PMID- 16241881 TI - The original Clarke Error Grid Analysis (EGA). PMID- 16241882 TI - Glycemic variability: it's not just about A1C anymore! PMID- 16241883 TI - Making sense of glucose monitoring technologies: from SMBG to CGM. PMID- 16241884 TI - Real-time continuous glucose monitor use and patient selection: what have we learned and where are we going? PMID- 16241885 TI - Hypoglycemia detection, and better yet, prevention, in pediatric patients. PMID- 16241886 TI - Reimbursement and coverage implications for CGM. PMID- 16241887 TI - New dawn for diabetes data management. PMID- 16241888 TI - Demonstrations and discussions of continuous glucose monitors. PMID- 16241889 TI - Incretins: what does the future hold? PMID- 16241890 TI - New insulin analogues. PMID- 16241891 TI - Intensive insulin treatment in type 2 diabetes. PMID- 16241892 TI - Practical considerations for glucose control in hospitalized patients. PMID- 16241894 TI - Pulmonary insulin update. PMID- 16241893 TI - Optimizing management of type 2 diabetes in hospitalized patients who are not critically ill. PMID- 16241895 TI - Hands-on demonstration and discussion of new pump software/hardware. PMID- 16241896 TI - To pump or not to pump. PMID- 16241897 TI - Comprehensive algorithm for quantitative real-time polymerase chain reaction. AB - Quantitative real-time polymerase chain reactions (qRT-PCR) have become the method of choice for rapid, sensitive, quantitative comparison of RNA transcript abundance. Useful data from this method depend on fitting data to theoretical curves that allow computation of mRNA levels. Calculating accurate mRNA levels requires important parameters such as reaction efficiency and the fractional cycle number at threshold (CT) to be used; however, many algorithms currently in use estimate these important parameters. Here we describe an objective method for quantifying qRT-PCR results using calculations based on the kinetics of individual PCR reactions without the need of the standard curve, independent of any assumptions or subjective judgments which allow direct calculation of efficiency and CT. We use a four-parameter logistic model to fit the raw fluorescence data as a function of PCR cycles to identify the exponential phase of the reaction. Next, we use a three-parameter simple exponent model to fit the exponential phase using an iterative nonlinear regression algorithm. Within the exponential portion of the curve, our technique automatically identifies candidate regression values using the P-value of regression and then uses a weighted average to compute a final efficiency for quantification. For CT determination, we chose the first positive second derivative maximum from the logistic model. This algorithm provides an objective and noise-resistant method for quantification of qRT-PCR results that is independent of the specific equipment used to perform PCR reactions. PMID- 16241898 TI - Structured motifs search. AB - In this paper, we describe an algorithm for the localization of structured models, i.e. sequences of (simple) motifs and distance constraints. It basically combines standard pattern matching procedures with a constraint satisfaction solver, and it has the ability, not present in similar tools, to search for partial matches. A significant feature of our approach, especially in terms of efficiency for the application context, is that the (potentially) exponentially many solutions to the considered problem are represented in compact form as a graph. Moreover, the time and space necessary to build the graph are linear in the number of occurrences of the component patterns. PMID- 16241899 TI - The statistical analysis of spatially clustered genes under the maximum gap criterion. AB - Statistical validation of gene clusters is imperative for many important applications in comparative genomics which depend on the identification of genomic regions that are historically and/or functionally related. We develop the first rigorous statistical treatment of max-gap clusters, a cluster definition frequently used in empirical studies. We present exact expressions for the probability of observing an individual cluster of a set of marked genes in one genome, as well as upper and lower bounds on the probability of observing a cluster of h homologs in a pairwise whole-genome comparison. We demonstrate the utility of our approach by applying it to a whole-genome comparison of E. coli and B. subtilis. Code for statistical tests is available at. PMID- 16241901 TI - Exact algorithms for planted motif problems. AB - The problem of identifying meaningful patterns (i.e., motifs) from biological data has been studied extensively due to its paramount importance. Three versions of this problem have been identified in the literature. One of these three problems is the planted (l, d)-motif problem. Several instances of this problem have been posed as a challenge. Numerous algorithms have been proposed in the literature that address this challenge. Many of these algorithms fall under the category of heuristic algorithms. In this paper we present algorithms for the planted (l, d)-motif problem that always find the correct answer(s). Our algorithms are very simple and are based on some ideas that are fundamentally different from the ones employed in the literature. We believe that the techniques we introduce in this paper will find independent applications. PMID- 16241900 TI - Genomic classification using an information-based similarity index: application to the SARS coronavirus. AB - Measures of genetic distance based on alignment methods are confined to studying sequences that are conserved and identifiable in all organisms under study. A number of alignment-free techniques based on either statistical linguistics or information theory have been developed to overcome the limitations of alignment methods. We present a novel alignment-free approach to measuring the similarity among genetic sequences that incorporates elements from both word rank order frequency statistics and information theory. We first validate this method on the human influenza A viral genomes as well as on the human mitochondrial DNA database. We then apply the method to study the origin of the SARS coronavirus. We find that the majority of the SARS genome is most closely related to group 1 coronaviruses, with smaller regions of matches to sequences from groups 2 and 3. The information based similarity index provides a new tool to measure the similarity between datasets based on their information content and may have a wide range of applications in the large-scale analysis of genomic databases. PMID- 16241902 TI - A construction of pooling designs with some happy surprises. AB - The screening of data sets for "positive data objects" is essential to modern technology. A (group) test that indicates whether a positive data object is in a specific subset or pool of the dataset can greatly facilitate the identification of all the positive data objects. A collection of tested pools is called a pooling design. Pooling designs are standard experimental tools in many biotechnical applications. In this paper, we use the (linear) subspace relation coupled with the general concept of a "containment matrix" to construct pooling designs with surprisingly high degrees of error correction (detection.) Error correcting pooling designs are important to biotechnical applications where error rates often are as high as 15%. What is also surprising is that the rank of the pooling design containment matrix is independent of the number of positive data objects in the dataset. PMID- 16241903 TI - Homologous versus heterologous interactions in the bicomponent staphylococcal gamma-haemolysin pore. AB - Staphylococcal gamma-haemolysin HlgA-HlgB forms a beta-barrel transmembrane pore in cells and in model membranes. The pore is formed by the oligomerization of two different proteins and a still debated number of monomers. To clarify the topology of the pore, we have mutated single residues - placed near the right and left interfaces of each monomer into cysteine. The mutants were labelled with fluorescent probes, forming a donor-acceptor pair for FRET (fluorescence resonance energy transfer). Heterologous couples (labelled on complementary left and right interfaces) displayed a marked FRET, suggesting extensive HlgA-HlgB or HlgB-HlgA contacts. Heterologous control couples (with both components labelled on the same side) showed absent or low FRET. We found the same result for the homologous couple formed by HlgA [i.e. HlgA-HlgA in the presence of wt (wild type) HlgB]. The homologous HlgB couple (HlgB-HlgB labelled on left and right interfaces and in the presence of wt HlgA) displayed a transient, declining FRET, which may indicate fast formation of an intermediate that is consumed during pore formation. We conclude that bicomponent pores are assembled by alternating heterologous monomers. PMID- 16241904 TI - Characterization of retinaldehyde dehydrogenase 3. AB - RALDH3 (retinal dehydrogenase 3) was characterized by kinetic and binding studies, protein engineering, homology modelling, ligand docking and electrostatic-potential calculations. The major recognition determinant of an RALDH3 substrate was shown to be an eight-carbon chain bonded to the aldehyde group whose kinetic influence (kcat/Km at pH 8.5) decreases when shortened or lengthened. Surprisingly, the b-ionone ring of all-trans-retinal is not a major recognition site. The dissociation constants (Kd) of the complexes of RALDH3 with octanal, NAD+ and NADH were determined by intrinsic tryptophan fluorescence. The similarity of the Kd values for the complexes with NAD+ and with octanal suggests a random kinetic mechanism for RALDH3, in contrast with the ordered sequential mechanism often associated with aldehyde dehydrogenase enzymes. Inhibition of RALDH3 by tri-iodothyronine binding in competition with NAD+, predicted by the modelling, was established kinetically and by immunoprecipitation. Mechanistic implications of the kinetically influential ionizations with macroscopic pKa values of 5.0 and 7.5 revealed by the pH-dependence of kcat are discussed. Analogies with data for non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase from Streptococcus mutans, together with the present modelled structure of the thioacyl RALDH3, suggest (a) that kcat characterizes deacylation of this intermediate for specific substrates and (b) the assignment of the pKa of the major ionization (approximating to 7.5) to the perturbed carboxy group of Glu280 whose conjugate base is envisaged as supplying general base catalysis to attack of a water molecule. The macroscopic pKa of the minor ionization (5.0) is considered to approximate to that of the carboxy group of Glu488. PMID- 16241905 TI - Plasma-membrane-associated sialidase (NEU3) differentially regulates integrin mediated cell proliferation through laminin- and fibronectin-derived signalling. AB - We have found previously that human plasma-membrane-associated sialidase (NEU3), a key glycosidase for ganglioside degradation, was markedly up-regulated in human colon cancers, with an involvement in suppression of apoptosis. To elucidate the molecular mechanisms underlying increased NEU3 expression, in the present study we investigated its role in cell adhesion of human colon cancer cells. DLD-1 cells transfected with NEU3 exhibited increased adhesion to laminins and consequent cell proliferation, but decreased cell adhesion to fibronectin and collagens I and IV, compared with control cells. When triggered by laminins, NEU3 clearly stimulated phosphorylation of FAK (focal adhesion kinase) and ERK (extracellular-signal-regulated kinase), whereas there was no activation on fibronectin. NEU3 markedly enhanced tyrosine phosphorylation of integrin beta4 with recruitment of Shc and Grb-2 only on laminin-5, and NEU3 was co immunoprecipitated by an anti-(integrin beta4) antibody, suggesting that association of NEU3 with integrin beta4 might facilitate promotion of the integrin-derived signalling on laminin-5. In addition, the promotion of phosphorylation of integrin beta1 and ILK (integrin-linked kinase) was also observed on laminins. G(M3) depletion as the result of NEU3 overexpression, assessed by TLC, appeared to be one of the causes of the increased adhesion on laminins and, in contrast, of the decreased adhesion on fibronectin - NEU3 probably having bimodal effects. These results indicate that NEU3 differentially regulates cell proliferation through integrin-mediated signalling depending on the extracellular matrix and, on laminins, NEU3 did indeed activate molecules often up-regulated in carcinogenesis, which may cause an acceleration of the malignant phenotype in cancer cells. PMID- 16241907 TI - PROactive Study: (r)evolution in the therapy of diabetes? PMID- 16241906 TI - Coupling of protein motions and hydrogen transfer during catalysis by Escherichia coli dihydrofolate reductase. AB - The enzyme DHFR (dihydrofolate reductase) catalyses hydride transfer from NADPH to, and protonation of, dihydrofolate. The physical basis of the hydride transfer step catalysed by DHFR from Escherichia coli has been studied through the measurement of the temperature dependence of the reaction rates and the kinetic isotope effects. Single turnover experiments at pH 7.0 revealed a strong dependence of the reaction rates on temperature. The observed relatively large difference in the activation energies for hydrogen and deuterium transfer led to a temperature dependence of the primary kinetic isotope effects from 3.0+/-0.2 at 5 degrees C to 2.2+/-0.2 at 40 degrees C and an inverse ratio of the pre exponential factors of 0.108+/-0.04. These results are consistent with theoretical models for hydrogen transfer that include contributions from quantum mechanical tunnelling coupled with protein motions that actively modulate the tunnelling distance. Previous work had suggested a coupling of a remote residue,Gly121, with the kinetic events at the active site. However, pre-steady state experiments at pH 7.0 with the mutant G121V-DHFR, in which Gly121 was replaced with valine, revealed that the chemical mechanism of DHFR catalysis was robust to this replacement. The reduced catalytic efficiency of G121V-DHFR was mainly a consequence of the significantly reduced pre-exponential factors, indicating the requirement for significant molecular reorganization during G121V DHFR catalysis. In contrast, steady-state measurements at pH 9.5, where hydride transfer is rate limiting, revealed temperature-independent kinetic isotope effects between 15 and 35 degrees C and a ratio of the pre-exponential factors above the semi-classical limit, suggesting a rigid active site configuration from which hydrogen tunnelling occurs. The mechanism by which hydrogen tunnelling in DHFR is coupled with the environment appears therefore to be sensitive to pH. PMID- 16241908 TI - Fat food for a bad mood. Could we treat and prevent depression in Type 2 diabetes by means of omega-3 polyunsaturated fatty acids? A review of the evidence. AB - AIMS: Evidence strongly suggests that depression is a common complication of Type 2 diabetes mellitus. However, there is considerable room to improve the effectiveness of pharmacological antidepressant agents, as in only 50-60% of the depressed subjects with diabetes does pharmacotherapy lead to remission of depression. The aim of the present paper was to review whether polyunsaturated fatty acids (PUFA) of the omega-3 family could be used for the prevention and treatment of depression in Type 2 diabetes. METHODS: MEDLINE database and published reference lists were used to identify studies that examined the associations between omega-3 PUFA and depression. To examine potential side effects, such as on glycaemic control, studies regarding the use of omega-3 supplements in Type 2 diabetes were also reviewed. RESULTS: Epidemiological and clinical studies suggest that a high intake of omega-3 PUFA protects against the development of depression. There is also some evidence that a low intake of omega 3 is associated with an increased risk of Type 2 diabetes, but the results are less conclusive. Results from randomized controlled trials in non-diabetic subjects with major depression show that eicosapentaenoic acid is an effective adjunct treatment of depression in diabetes, while docosahexanoic acid is not. Moreover, consumption of omega-3 PUFA reduces the risk of cardiovascular disease and may therefore indirectly decrease depression in Type 2 diabetes, via the reduction of cardiovascular complications. CONCLUSIONS: Supplementation with omega-3 PUFA, in particular eicosapentaenoic acid, may be a safe and helpful tool to reduce the incidence of depression and to treat depression in Type 2 diabetes. Further studies are now justified to test these hypotheses in patients with Type 2 diabetes. PMID- 16241909 TI - Are insulin resistance, impaired fasting glucose and impaired glucose tolerance all equally strongly related to age? AB - BACKGROUND: Insulin resistance (IR) has been considered an underlying cause of impaired glucose tolerance (IGT) and impaired fasting glucose (IFG). Whether IR increases with age has been debated. We investigated the age-associated deterioration in the homeostasis model assessment (HOMA) of IR and in glucose metabolism. METHODS: Ten (nine including women) European studies contributed data on 6314 men and 6393 women aged 30-88 years. The cohort- and sex-specific top 25% of HOMA of IR in non-diabetic subjects was used to define HOMA-IR. RESULTS: Compared with subjects aged 50-59 years, the cohort- and body mass index-adjusted odds ratio (95% confidence interval) for HOMA-IR was 0.83 (0.64, 1.08), 0.87 (0.74, 1.03), 1.20 (1.02, 1.42) and 1.45 (1.10, 1.92) in men and 0.84 (0.62, 1.14), 0.91 (0.77, 1.09), 1.38 (1.19, 1.62) and 1.71 (1.35, 2.17) in women, respectively, aged 30-39, 40-49, 60-69 and > or = 70 years (P < 0.0001 for trend test). The same increasing trend was also observed for IFG. In contrast, the corresponding odds ratios for IGT increased linearly and more strongly with age, being 0.37 (0.22, 0.63), 0.67 (0.52, 0.87), 1.55 (1.24, 1.92) and 2.96 (2.13, 4.13) in men and 0.51 (0.31, 0.85), 0.66 (0.52, 0.86), 1.92 (1.57, 2.35) and 3.85 (2.89, 5.12) in women, respectively. CONCLUSIONS: Age is more strongly associated with IGT than with HOMA-IR or IFG in non-diabetic European populations. PMID- 16241910 TI - Evaluation of the association between the EQ-5D (health-related utility) and body mass index (obesity) in hospital-treated people with Type 1 diabetes, Type 2 diabetes and with no diagnosed diabetes. AB - AIMS: The purpose of this study was to characterize the impact of body mass index (BMI) on health-related utility for patients with Type 1 and Type 2 diabetes and those without diabetes. METHODS: The study was conducted in Cardiff and the Vale of Glamorgan, Wales, UK. Health-related utility was measured using the EQ 5D(index). Patients from the Health Outcomes Data Repository (HODaR) were surveyed by postal questionnaire either 6 weeks post discharge for in-patients or at out-patient clinics between January 2002 and July 2003. BMI was calculated from self-reported data within the survey. Patients with diabetes were identified by a previous history of an in-patient admission with diabetes or as an out patient with diabetes recorded as a coexisting diagnosis. RESULTS: Questionnaires were returned from 27 924 patients of whom 2575 had diabetes. Increasing BMI was found to reduce utility in all three groups. BMI was significantly greater for those with Type 2 diabetes compared with those with Type 1, and those without diabetes (P < 0.001). Multiple regression analysis demonstrated that both BMI and diabetes status had a significant effect on utility. However, the rate of change of utility attributable to BMI was not found to be significantly different between the various groups. CONCLUSIONS: Obesity negatively impacts upon health related utility and thus quality of life for all patient groups. There was no significant difference in the effect of obesity on utility between those with and without diabetes. PMID- 16241911 TI - Pilot study of the effects of local pressure on microvascular function in the diabetic foot. AB - AIM: The aim of the study was to determine whether areas of the diabetic foot that experience high pressures during normal activity also demonstrate reductions in cutaneous microvascular flow and/or endothelial function. METHODS: Sixteen patients with diabetes mellitus and eight healthy, age-matched control subjects were recruited. Maps of dynamic pressure on the plantar aspect of both feet were recorded during a normal gait cycle, and the skin microvascular blood flow response to the endothelium-dependent vasodilator acetylcholine was assessed at the sites of highest and lowest plantar pressure over the metatarsal heads. RESULTS: Patients with diabetes had higher plantar pressures than control subjects (P = 0.002), but there were no significant differences in basal skin blood flow or acetylcholine response between the groups. In the patients, baseline flow was increased (P = 0.041) but the acetylcholine response reduced (P = 0.03) at the high-pressure compared with the low-pressure site; this was most apparent in those who were particularly at risk of ulceration due to high plantar pressures. CONCLUSIONS: Chronically raised plantar pressure in the diabetic foot is associated with increased basal skin blood flow, compared with lower pressure areas on the same foot. Further work is required to determine whether, and under what conditions, this additional hyperaemia is protective or maladaptive. In addition, high-pressure areas have a reduced responsiveness to an endothelium dependent vasodilator, although the clinical significance of these changes is not clear. PMID- 16241912 TI - Rural Vincentians' (Caribbean) beliefs about the usage of non-prescribable medicines for treating Type 2 diabetes. AB - AIMS: To explore beliefs among persons in St. Vincent, a rural Caribbean Community, regarding the usage of non-prescribable medicines for treating Type 2 diabetes. METHODS: A phenomenological enquiry explored people's experiences and the manner in which they interpret these. One focus-group interview was conducted of persons attending a rural diabetes clinic in St Vincent to generate insights into the phenomenon. Analysis was undertaken using the four steps utilized in phenomenological studies of bracketing, intuiting, analysis and description. The findings result in deeper understanding and definition of the phenomenon. RESULTS: A variety of non-prescribable, predominantly herbal and folk, medicines were commonly used as a means of self care in diabetes. Usage was underpinned by a system of lay beliefs about diabetes and beliefs in the treatment efficacy of folk medicine. A strong religious influence formed the basis of diabetes treatment and offered some symptom relief and therefore treatment satisfaction through spiritual revelations about remedies. Conventional medicines were taken in conjunction with non-prescribable treatments or else not at all. CONCLUSIONS: Non-prescribable medicines were believed by participants to be efficacious. Conventional medication was perceived as an access to medical care. Study findings may be relevant to other rural populations with strong social and religious mores. PMID- 16241913 TI - Diabetes screening anxiety and beliefs. AB - AIMS: This study assesses the impact of screening for diabetes on anxiety levels in an ethnically mixed population in the UK, and explores whether beliefs about Type 2 diabetes account for these anxiety levels. METHODS: This cross-sectional study recruited individuals who were identified at high risk of developing diabetes through general practitioners' (GPs) lists or through public media recruitment. Participants completed an oral glucose tolerance test (OGTT). Between blood tests, participants completed the Spielberger State Anxiety Scale Short Form, the Emotional Stability Scale of the Big Five Inventory 44 and three scales from the Diabetes Illness Representations Questionnaire, revised for this study. RESULTS: Of the 1339 who completed the OGTT and questionnaire booklet, 54% were female, with 21% from an Asian background. Forty-five per cent of participants reported little to moderate amounts of anxiety at screening (mean 35.2; sd = 11.6). There was no significant effect of family history of diabetes, ethnic group or recruitment method on anxiety. The only variable significantly associated (negatively) with anxiety was the personality trait of emotional stability. Of responders, 64% and 61% agreed that diabetes was caused by diet or hereditary factors, respectively. Only 155 individuals (12%) agreed that diabetes was serious, shortens life and causes complications. CONCLUSIONS: The results of this study replicate that of previous studies, indicating that screening for diabetes does not induce significant anxiety. Bivariate analysis indicated that individuals who perceived diabetes to be serious, life shortening and resulting in complications had higher anxiety scores, the personality trait of emotional stability being the strongest predictor of anxiety. PMID- 16241914 TI - Is pregnancy a risk factor for microvascular complications? The EURODIAB Prospective Complications Study. AB - AIMS: To examine the long-term influence of pregnancy on the development and progression of microvascular complications in Type 1 diabetes. METHODS: In the EURODIAB Prospective Complications Study (PCS), 793 women potentially child bearing at baseline completed the follow-up (7.3 years) and 163 (21%) gave birth during the follow-up period. We compared risk factors [mean levels of age, duration of diabetes, HbA(1c), systolic blood pressure (SBP) and proportion giving birth] between those that did or did not develop microvascular complications during the follow-up period. RESULTS: For the 425 childless women at baseline, 102 gave birth during follow-up. HbA(1c) was a significant risk factor for progression to microalbuminuria but age, duration of diabetes, systolic blood pressure or giving birth were not. Duration of diabetes and high HbA(1c) were significant risk factors for progression to proliferative retinopathy, whereas giving birth was not. Similar results were obtained for progression to any form of retinopathy. Giving birth was not significantly related to the incidence of neuropathy. Similar results were obtained for women with children at baseline giving birth during follow-up (n = 61/368). CONCLUSIONS: In this European study, having a first or another pregnancy did not seem to be a risk factor for long-term progression of any microvascular complication. This is in accordance with the findings of the Diabetes Control and Complications Trial (DCCT). PMID- 16241915 TI - Synergism between mutant HNF1A and the metabolic syndrome in Oji-Cree Type 2 diabetes. AB - AIMS: To determine the prevalence of the metabolic syndrome in the Sandy Lake Oji Cree and to examine its interaction with HNF1A in association with impaired glucose tolerance and Type 2 diabetes. METHODS: Using data collected from the Sandy Lake Health and Diabetes Project (1993-1995), the presence or absence of the metabolic syndrome was determined in 515 Oji-Cree subjects, > or = 18 years of age. In the original study, fasting plasma analytes were measured, a 75-g oral glucose tolerance test was administered, and subjects were genotyped for HNF1A G319S. RESULTS: The unadjusted prevalence of the metabolic syndrome in the Oji Cree adults was 29.9%. The adjusted odds ratio (OR) and 95% confidence interval for Type 2 diabetes among subjects who carried the HNF1A G319S mutation and had the modified metabolic syndrome (excluding hyperglycaemia) was 20.3 (6.94, 59.6). Adjusted ORs for Type 2 diabetes for subjects with either the HNF1A G319S mutation alone or the modified metabolic syndrome alone were 5.56 (2.85, 10.9) and 4.84 (2.53, 9.27), respectively. The risk of having impaired glucose tolerance was not influenced by the presence of either factor. CONCLUSIONS: The risk of Type 2 diabetes was similar (approximately five-fold increased) for subjects with either the presence of the modified metabolic syndrome or the private HNF1A G319S mutation. Interestingly, when present in combination, the two independent risk factors appeared to act synergistically to confer an even greater increased risk of Type 2 diabetes. PMID- 16241916 TI - Peroxisome proliferator-activated receptor-gamma co-activator-1alpha (PGC-1alpha) gene polymorphisms and their relationship to Type 2 diabetes in Asian Indians. AB - AIMS: The objective of the present investigation was to examine the relationship of three polymorphisms, Thr394Thr, Gly482Ser and +A2962G, of the peroxisome proliferator activated receptor-gamma co-activator-1 alpha (PGC-1alpha) gene with Type 2 diabetes in Asian Indians. METHODS: The study group comprised 515 Type 2 diabetic and 882 normal glucose tolerant subjects chosen from the Chennai Urban Rural Epidemiology Study, an ongoing population-based study in southern India. The three polymorphisms were genotyped using polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). Haplotype frequencies were estimated using an expectation-maximization (EM) algorithm. Linkage disequilibrium was estimated from the estimates of haplotypic frequencies. RESULTS: The three polymorphisms studied were not in linkage disequilibrium. With respect to the Thr394Thr polymorphism, 20% of the Type 2 diabetic patients (103/515) had the GA genotype compared with 12% of the normal glucose tolerance (NGT) subjects (108/882) (P = 0.0004). The frequency of the A allele was also higher in Type 2 diabetic subjects (0.11) compared with NGT subjects (0.07) (P = 0.002). Regression analysis revealed the odds ratio for Type 2 diabetes for the susceptible genotype (XA) to be 1.683 (95% confidence intervals: 1.264-2.241, P = 0.0004). Age adjusted glycated haemoglobin (P = 0.003), serum cholesterol (P = 0.001) and low-density lipoprotein (LDL) cholesterol (P = 0.001) levels and systolic blood pressure (P = 0.001) were higher in the NGT subjects with the XA genotype compared with GG genotype. There were no differences in genotype or allelic distribution between the Type 2 diabetic and NGT subjects with respect to the Gly482Ser and +A2962G polymorphisms. CONCLUSIONS: The A allele of Thr394Thr (G --> A) polymorphism of the PGC-1 gene is associated with Type 2 diabetes in Asian Indian subjects and the XA genotype confers 1.6 times higher risk for Type 2 diabetes compared with the GG genotype in this population. PMID- 16241917 TI - Evaluation of a programme of group visits and computer-assisted consultations in the treatment of adolescents with Type 1 diabetes. AB - AIM: To examine the effects of group visits and computer-assisted consultations on quality of life and glycaemic control in adolescents with Type 1 diabetes. METHODS: A total of 116 adolescents, aged 11-17 years, and their parents were randomly assigned to an intervention (n = 62) or a control group (n = 54). The intervention group was invited to a 15-month programme comprising group visits and computer-assisted consultations. The control group was offered traditional out-patient consultations. Outcomes included changes in HbA(1c) and the adolescents' assessment of generic and disease-specific health-related quality of life measured by the Child Health Questionnaire (CHQ-CF87) and the Diabetes Quality of Life Questionnaire (DQOL), respectively. RESULTS: One hundred and one adolescents (55/46) agreed to participate, mean age 14.2 years (sd 1.5), mean diabetes duration 6.5 years (sd 3.6, range 1-16 years), mean HbA(1c) 9.3% (sd 1.4, range 6.1-12.8%). Eighty-three (72%) completed the questionnaires at follow up (intervention/control 45/38). There were significant age by randomization group interactions for diabetes-related impact (P = 0.018), diabetes-related worries (P = 0.004), mental health (P = 0.046) and general behaviour (P = 0.029), implying that the intervention was effective in older adolescents (above 13-14 years). No significant effects on mean HbA(1c) were identified. CONCLUSIONS: Group visits and computer-assisted consultations had beneficial effects on health related quality of life in older adolescents, the role of this intervention being questionable in younger adolescents. PMID- 16241918 TI - Prognostic significance of glycaemic control in patients with HBV and HCV-related cirrhosis and diabetes mellitus. AB - AIMS: Diabetes mellitus (DM) is frequently observed in patients with cirrhosis, particularly that due to hepatitis C virus (HCV) infection. However, no studies have focused on the clinical significance of glycaemic control in cirrhotic patients because of their short life expectancy and poor hepatic function. The aim of this study was to evaluate the prognostic impact of glycaemic control in patients with hepatitis B virus (HBV) and HCV-related cirrhosis and DM. METHODS: A total of 434 patients with HCV-related (HCV group, n = 88) or HBV-related (HBV group, n = 346) cirrhosis were studied retrospectively. We determined the prevalence of DM and treatment methods for hyperglycaemia and status of glycaemic control, and the patients' outcome. RESULTS: The prevalence of DM was 43.2% (38/88) in the HCV group and 19.7% (68/346) in the HBV group. Patients in the HCV group were older with a female preponderance. DM was detected before the diagnosis of cirrhosis or simultaneously in 92% and 79% in the HCV and HBV groups, respectively. Most patients were treated with insulin or oral hypoglycaemic agents. However, blood glucose levels were maintained within the normal range in 34.2% of the HCV group and in 23.5% of the HBV group. Forty-six patients died during the observation period in both groups. Hepatic failure was the most common cause of death, and sepsis and variceal bleeding were more frequent in the HCV group than in the HBV group. Multivariate analysis showed that Child-Pugh class was the most important factor for survival in both groups. In the HCV group, the status of glycaemic control was a significant independent factor of survival (P = 0.018). In the HBV group, age and the development of spontaneous bacterial peritonitis were significant. CONCLUSION: DM is more frequent in patients with HCV-related cirrhosis than in patients with HBV. Strict control of blood glucose levels could improve survival in HCV patients. A precise assessment of the risks and benefits of glycaemic control is required to reduce the mortality and morbidity of patients with cirrhosis and DM. PMID- 16241919 TI - Evaluating the therapeutic approach in pregnancies complicated by borderline glucose intolerance: a randomized clinical trial. AB - AIMS: Most studies relating minor gestational metabolic alterations to macrosomia refer to glucose intolerance classified on the basis of the National Diabetes Data Group or previous World Health Organization diagnostic thresholds. Our aim was to evaluate the consequences of very mild forms of gestational glucose intolerance, defined by an elevated 50-g glucose challenge test followed by a normal oral glucose tolerance test, using the more restrictive Carpenter and Coustan's criteria (Borderline Gestational Glucose Intolerance, BGGI). METHODS: Three hundred BGGI women were randomly assigned to: Group A (standard management), Group B (dietary treatment and regular monitoring). A control group (C) was also considered. Newborns were classified as macrosomic, large (LGA), or small for gestational age (SGA). RESULTS: The three groups were similar in age, body mass index and parity. Therapy in Group B significantly improved fasting (from 4.68 +/- 0.45 to 4.28 +/- 0.45 mmol/l) and 2-h postprandial glycaemia (from 6.01 +/- 0.57 to 5.13 +/- 0.68 mmol/l). Fasting glycaemia at delivery was significantly lower in B (4.20 +/- 0.38 mmol/l) than in A (4.84 +/- 0.45 mmol/l), and was also lower than in C (4.31 +/- 0.39 mmol/l). Significantly fewer LGA babies were born to Group B (6.0%) than Group A (14.0%) and Group C (9.1%). No difference was found in the SGA rate. The neonatal Ponderal Index was higher (P = 0.030) in group A (2.73 +/- 0.35) than in C (2.64 +/- 0.30) and B (2.64 +/- 0.24). CONCLUSIONS: Even very mild alterations in glucose tolerance can result in excessive or disharmonious fetal growth, which may be prevented by simple, non invasive therapeutic measures. PMID- 16241920 TI - Diabetes mellitus: clinical presentation and outcome in men and women with acute coronary syndromes. Data from the Euro Heart Survey ACS. AB - AIMS: To study clinical presentation, in-hospital course and short-term prognosis in men and women with diabetes mellitus and acute coronary syndromes (ACS). METHODS: Men (n = 6488, 21.2% with diabetes) and 2809 women (28.7% with diabetes) < or = 80 years old, with a discharge diagnosis of ACS were prospectively enrolled in the Euro Heart Survey of ACS. RESULTS: Women with diabetes were more likely to present with ST elevation than non-diabetic women, a difference that became more marked after adjustment for differences in smoking, hypertension, obesity, medication and prior disease [adjusted odds ratio (OR) 1.46 (1.20, 1.78)], whereas there was little difference between diabetic and non-diabetic men [adjusted OR 0.99 (0.86, 1.14)]. In addition, women with diabetes were more likely to develop Q-wave myocardial infarction (MI) than non-diabetic women [adjusted OR 1.61 (1.30, 1.99)], while there was no difference between men with and without diabetes [adjusted OR 0.99 (0.85, 1.15)]. There were significant interactions between sex, diabetes and presenting with ST-elevation ACS (P < 0.001), and Q-wave MI (P < 0.001), respectively. Of the women with diabetes, 7.4% died in hospital, compared with 3.6% of non-diabetic women [adjusted OR 2.13 (1.39, 3.26)], whereas corresponding mortality rates in men with and without diabetes were 4.1% and 3.3%, respectively [OR 1.13 (0.76, 1.67)] (P for diabetes sex interaction 0.021). CONCLUSION: In women with ACS, diabetes is associated with higher risk of presenting with ST-elevation ACS, developing Q-wave MI, and of in-hospital mortality, whereas in men with ACS diabetes is not significantly associated with increased risk of either. These findings suggest a differential effect of diabetes on the pathophysiology of ACS based on the patient's sex. PMID- 16241921 TI - Increased risk of diabetes among relatives of female insulin-treated patients diagnosed at 15-34 years of age. AB - AIMS: This study aimed to determine the risk of developing diabetes among relatives of patients diagnosed between 15 and 34 years of age who were treated with insulin. Our second aim was to determine whether there was a difference in risk of diabetes between relatives of male and female patients. METHODS: A questionnaire was sent to patients in the Diabetes Incidence Study in Sweden registry diagnosed between 1983 and 1993 to determine the presence of first degree relatives with diabetes. RESULTS: In 3087 index patients treated with insulin, 17.8% (95% confidence interval 16.5, 19.2) had a first-degree relative (excluding offspring) treated with insulin, the frequency being higher in female (19.8%) than in male (16.5%, P = 0.018) patients. A total of 10.7% had a parent treated with insulin. The prevalence of insulin-treated diabetes was higher among parents of female (12.5%) than of male (9.5%), insulin-treated index patients (P = 0.0068). A similar difference was observed using life table analysis (P = 0.0025), which also showed that the risk by 63 years of age was 7.6% for parents of female and 4.9% for parents of male insulin-treated index patients. In insulin treated index patients, 8.4% had a sibling with insulin-treated diabetes. CONCLUSIONS: We conclude that the risk for relatives of women with insulin treated diabetes was higher than for relatives of insulin-treated male patients. We suggest that greater genetic susceptibility is required for females compared with males in the 15-34 age group in order to develop diabetes and hence females might carry more diabetes genes since more of their relatives also develop diabetes. PMID- 16241922 TI - Urinary albumin levels in the normal range determine arterial wall thickness in adults with Type 2 diabetes: a FIELD substudy. AB - AIM: Cardiovascular disease (CVD) rates are substantially higher among patients with Type 2 diabetes than in the general population. The objective of this study was to identify the determinants of carotid intima media thickness (IMT) in patients with Type 2 diabetes. METHODS: We measured the thickness of the intima media layer of the carotid artery, a strong predictor of the risk of future vascular events, in 397 Type 2 diabetic patients drawn from the Fenofibrate Intervention and Event Lowering in Diabetes study, prior to treatment allocation. RESULTS: The mean IMT was 0.78 mm [interquartile range (IQR) 0.23 mm], and the maximum IMT was 1.17 mm (IQR 0.36 mm). By multivariate analysis, age, sex, duration of diabetes, triglycerides, and total cholesterol were independently correlated with IMT, as was urine albumin-creatinine ratio (ACR) (P < 0.001). The effect of ACR on IMT was further examined by tertile. Clinically significant differences in IMT were associated with ACR > 0.65 mg/mmol, approximately one fifth the standard clinical threshold for microalbuminuria (P < 0.01). Long-term diabetes, independent of other parameters, was associated with a 50% increase in age-related thickening. CONCLUSIONS: IMT in people with Type 2 diabetes is independently and continuously related to urine albumin levels and to the duration of diabetes. These results support previous data linking urine albumin measurements within the normal range with increased ischaemic cardiac mortality in the setting of Type 2 diabetes, and strongly suggest that urine albumin levels within this range should trigger a formal evaluation for CVD. PMID- 16241923 TI - Factors associated with glycaemic outcome of childhood diabetes care in Denmark. AB - AIMS: To study how structure and process of care is associated with outcome assessed by HbA(1c). METHODS: Data for this cross-sectional study originated from the nationwide Danish Registry for Childhood Diabetes and two questionnaires. One questionnaire was sent to all children under 16 years of age with Type 1 diabetes in the year 2000 (N = 1087, response rate 80%). Another questionnaire was sent to the 19 centres in Denmark treating these children (response rate 100%). Simultaneously the children were asked to take a blood sample for central HbA(1c) analysis. Linear mixed models were used for analysis of associations between structure and process indicators and HbA(1c). Age, diabetes duration, sex, ethnicity, family structure and parents' occupational status were included as patient factors possibly affecting HbA(1c). RESULTS: More visits to the outpatient clinic and higher insulin dosage were significantly associated with higher HbA(1c) (P = 0.002 and P = 0.0001, respectively). Increased frequency of blood glucose monitoring (BGM/week) and completed nephropathy screening were significantly associated with lower HbA(1c) value (estimates -0.008 and -0.49, P = 0.02, respectively). The structure indicators were not associated with HbA(1c), but telephone hot-line was positively associated with the process indicator BGM (estimate 4.02, P = 0.04). Children without Danish parents performed BGM significantly less frequently (-7.11, P = 0.0005) and had higher HbA(1c) (0.41, P = 0.06). CONCLUSIONS: Most process indicators were significantly associated with HbA(1c), indicating relevant action of staff on glucose regulation. The structure indicators were not associated with outcome, necessitating more detailed studies on the influence of staffing resources, treatment strategies and targets in childhood diabetes management. PMID- 16241924 TI - Does parity increase insulin resistance during pregnancy? AB - AIMS: To study the effect of parity on impairment of insulin sensitivity during pregnancy and on the risk of gestational diabetes (GDM). METHODS: We studied the relationship between parity and peripheral insulin sensitivity index (ISI(OGTT)) or GDM in 1880 caucasian women, who underwent a 100-g, 3-h oral glucose tolerance test (OGTT) between the 24th and 28th gestational week and in 75 women who underwent an OGTT in two consecutive pregnancies. A proxy for beta-cell function (basal plasma C peptide/fasting plasma glucose; CP/FPG) was also measured. RESULTS: By univariate analysis parity was related to decreased ISI(OGTT) and to increased CP/FPG in those with parity > 3 and likewise GDM, diagnosed in 124 women (6.58%), was linearly related to parity (P = 0.0034) and strongly age dependent. The relationships between parity and ISI(OGTT), CP/FPG and GDM were no longer significant after adjustment for age, pregestational body mass index (BMI), and weight gain. GDM was significantly related to age and pregestational weight, while ISI(OGTT) and CP/FPG were inversely related to prepregnancy BMI or weight gain. In comparison with the index pregnancy, the subsequent pregnancy was characterized by an increase in actual and prepregnancy BMI, in 2 h area under curve (AUC) glucose and by a decrease in ISI(OGTT) (P = 0.0001). The longer the time interval between pregnancies and the higher the increment in pregestational BMI or in weight gain during the pregnancy, the greater were the ISI(OGTT) decrease and 2-h AUC glucose increase. CONCLUSIONS: Parity is not directly linked to insulin sensitivity deterioration, to CP/FPG increase during pregnancy, or to GDM appearance, although it is linked through the mediation of progressive ageing and weight gain either before or during pregnancy, when there is a sufficiently long time interval between pregnancies. PMID- 16241925 TI - The epidemiology of Type 2 diabetes mellitus in Greek adults: the ATTICA study. AB - AIMS: Individuals with Type 2 diabetes are at high risk for coronary heart disease, and may benefit from aggressive lifestyle modification. We evaluated the prevalence of Type 2 diabetes, in a Greek adult population, in relation to physical activity and dietary habits. METHODS: From May 2001 to December 2002, we randomly enrolled 1514 men and 1528 women, with no evidence of cardiovascular or any other chronic disease. The sampling was stratified by the age and gender distribution of the general population in the greater area of Athens (census 2001). Diabetes was defined according to the established American Diabetes Association criteria. Dietary habits were assessed through a validated food frequency questionnaire and a diet score was developed, in which higher values suggest greater adherence to the Mediterranean diet. Weekly energy expenditure was assessed by considering frequency, duration and intensity of sports-related physical activity during a usual week. RESULTS: After age adjustment for the Greek adult population (2001 census), the projected prevalence of Type 2 diabetes was 7.6% in men and 5.9% in women. A significant proportion of diabetic subjects (24% of men and 30% of women) were unaware of their condition. Moreover, a 10 unit increase in the diet score was associated with 21% lower odds of diabetes (P < 0.05), while individuals taking light physical activity were at 35% lower odds ratio of diabetes compared with sedentary individuals (P < 0.05). CONCLUSIONS: Diabetes mellitus is common in our population, with approximately 1 in 5 individuals with Type 2 diabetes being unaware of their condition. The lifestyle approach described here may contribute significantly to the reduction in the prevalence of diabetes, at a population level. PMID- 16241926 TI - The metabolic syndrome and changing relationship between blood pressure and insulin with age, as observed in Aboriginal and Torres Strait Islander peoples. AB - AIMS: To determine the prevalence of the metabolic syndrome (MS) among Aboriginal and Torres Strait Islander peoples. A further objective was to investigate the relationships between fasting insulin and blood pressure (BP) within these groups with increasing age. METHODS: A cross-sectional population-based study included 369 Torres Strait Islanders (residing in Torres Strait and Far North Queensland), and 675 Aborigines from central Australia. Data necessary for classification of MS was collected, including fasting and 2-h glucose and insulin, urinary albumin and creatinine, anthropometric measurements, BP, serum lipids. RESULTS: The ATPIII criteria classified 43% of Torres Strait Islanders and 44% of Aborigines with MS, whereas 32 and 28%, respectively, had the MS according to WHO criteria. Agreement between the two criteria was only modest (kappa coefficient from 0.28 to 0.57). Factor analyses indicated no cluster including both insulin and BP in either population. Significant correlations (P < 0.05) [adjusted for gender, body mass index (BMI) and waist circumference] were observed between BP and fasting insulin: a positive correlation for Torres Strait Islanders aged 15-29 years, and an inverse correlation for Aborigines aged 40 years and older. CONCLUSION: Torres Strait Islanders and Aborigines had very high prevalences of the MS. Specific population characteristics (high prevalences of central obesity, dyslipidaemia, renal disease) may make the WHO definition preferable to the ATPIII definition in these population groups. The poor agreement between criteria suggests a more precise definition of the metabolic syndrome that is applicable across populations is required. This study showed an inverse relationship with age for the correlation of BP and fasting insulin. PMID- 16241927 TI - Ethnic differences in glycaemic control in adult Type 2 diabetic patients in primary care: a 3-year follow-up study. AB - OBJECTIVE: To evaluate ethnic differences and characteristics related to glycaemic control in patients with Type 2 diabetes in primary care. RESEARCH DESIGN AND METHODS: Prospective cohort study; 500 adult patients with Type 2 diabetes, who were not on insulin therapy, were followed up annually for 3 years. HbA(1c) at baseline and 3-year changes and subsequent insulin therapy were related to baseline characteristics. RESULTS: Malay patients had significantly higher HbA(1c) (mean 8.7% +/- sd 1.66) compared with Chinese (8.2 +/- sd 1.67) and Indian (8.2 +/- sd 1.55) (P = 0.032) at baseline, and consistently for all years of HbA(1c) assessment (P = 0.017). At baseline, Malay patients were significantly more obese than Chinese or Indians (P < 0.001); fewer of them received structured shared-care intervention (P = 0.001), but they had a significantly higher glucose control educational score (P < 0.05). Multivariable analyses showed that HbA(1c) at baseline was significantly related to age (P = 0.001), BMI (P = 0.031) and ethnicity (P = 0.002). HbA(1c) declined significantly over 3 years in the whole population and in all ethnic groups. Significantly greater HbA(1c) declines were associated with higher baseline HbA(1c), structured shared-care intervention and non-insulin therapy. Correcting for differences on these factors, the decline in HbA(1c) in Malays was significantly less than in the Chinese. Insulin therapy was associated with higher baseline HbA(1c) and higher BMI. CONCLUSIONS: Malay ethnicity was associated with persistently poor glycaemic control. Sociocultural and behavioural factors should be addressed in improving care for patients with poorly controlled diabetes. PMID- 16241928 TI - Life-threatening hypokalaemia on a low-carbohydrate diet associated with previously undiagnosed primary hyperaldosteronism [corrected]. AB - BACKGROUND: Low-carbohydrate diets are popular and fashionable for weight loss despite lack of evidence about long-term effects. Many individuals attempting to lose weight have hypertension, especially those with diabetes, and the prevalence of hyperaldosteronism among hypertensive patients is higher than previously recognized. We present a patient with Type 2 diabetes and previously undiagnosed hyperaldosteronism who developed life-threatening hypokalaemia while following a low-carbohydrate diet. CASE REPORT: A 60-year-old man with diet-treated Type 2 diabetes and hypertension presented with generalized muscle weakness and serum potassium of 1.9 mmol/l. He had succeeded in losing three and a half stones during the previous 4 months by adhering strictly to a low-carbohydrate diet. HbA(1c) was 4.8% and plasma aldosterone:renin ratios were elevated suggestive of increased aldosterone secretion. On a low-calorie mixed diet serum potassium levels were maintained in the low-normal range over the following 165 days. The adrenals were normal on CT scanning and blood pressure responded dramatically to the addition of spironolactone on day 212 (125/83 mmHg). CONCLUSIONS: The prevalence of primary hyperaldosteronism in the hypertensive population, based on elevation of plasma renin:aldosterone ratio, is approximately 6%. The majority of these people are normokalaemic and remain undiagnosed. However, when carbohydrate intake is restricted such individuals are at increased risk of potentially life threatening metabolic derangements. PMID- 16241929 TI - Pseudoclaudication as a manifestation of diabetic neuropathy. AB - We present the case of a 64-year-old male Type 1 diabetic patient with painful diabetic neuropathy masquerading as intermittent claudication. Examination of the peripheral circulation (both arterial and venous) was normal. An MRI scan excluded lumbar spinal stenosis and nerve root compression as the cause of claudication. The case suggests that, in the absence of other identifiable causes and in the presence of peripheral diabetic neuropathy, "intermittent claudication" may be due to the neuropathy itself. PMID- 16241930 TI - Long-term treatment experience in a subject with Dunnigan-type familial partial lipodystrophy: efficacy of rosiglitazone. AB - Dunnigan-type familial partial lipodystrophy (FPLD) is caused by mutations in LMNA, the gene that encodes nuclear lamins A and C. FPLD is characterized by peripheral fat loss, excess central adiposity, insulin resistance, and hyperlipidaemia, which are difficult to treat. We present our 2 years' experience of treatment with rosiglitazone in a subject with FPLD. Insulin requirement decreased significantly from 240 IU/day to 76 IU/day (range 20-240 IU/day) and serum triglyceride concentration was lowered from 13.7 +/- 14.4 mmol/l to 4.5 +/- 4.3 mmol/l and remained stable. Mean HbA(1c) prior to rosiglitazone therapy was 9.4 +/- 1.32% and decreased to 7.4 +/- 0.6% during therapy with rosiglitazone. This case demonstrates the benefits of PPARgamma-agonists on glycaemic control and dyslipidaemia in a patient with FPLD. This in turn implies that PPARgamma may play a pathophysiological role in FPLD. PMID- 16241931 TI - Coronary flow reserve, insulin resistance and blood pressure response to standing in patients with normoglycaemia: is there a relationship? AB - AIMS: To establish the relationships between coronary flow reserve, cardiovascular autonomic function, and insulin resistance characterized by the homeostasis model assessment insulin resistance score in patients with normal carbohydrate metabolism according to the World Health Organization (WHO) and American Diabetes Association (ADA) criteria, and with morphologically normal epicardial coronary arteries. METHODS: Twenty-five patients [12 women and 13 men, mean (sd) age: 53 +/- 11 years] with normal coronary angiography were enrolled into the study. Coronary flow reserve was measured during stress transoesophageal echocardiography. Autonomic dysfunction was assessed by means of five standard cardiovascular reflex tests. The fasting serum glucose and insulin levels were determined and the homeostasis assessment model insulin resistance score was calculated. RESULTS: In patients with normal carbohydrate metabolism, negative correlations were observed between the coronary flow reserve and both the serum insulin level (r = -0.445, P = 0.026) and the homeostasis assessment model insulin resistance score (r = -0.449, P = 0.024). The systolic blood pressure response to standing also correlated with the coronary flow reserve (r = -0.519, P = 0.011). The heart rate response to deep breathing, the Valsalva ratio, the 30/15 ratio and the sustained handgrip test results were not correlated with the coronary flow reserve. CONCLUSIONS: Our data suggest the possible role of insulin resistance and early sympathetic nerve dysfunction in the development of decreased coronary flow reserve in patients without diabetes mellitus or impaired glucose tolerance. PMID- 16241932 TI - Short-term effects of cognitive behavioural group training (CBGT) in adult Type 1 diabetes patients in prolonged poor glycaemic control. A randomized controlled trial. AB - AIMS: To assess the effects of cognitive behavioural group training (CBGT) on glycaemic control, diabetes self-efficacy and well-being in Type 1 diabetes patients in persistent poor glycaemic control. METHODS: In a randomized controlled trial, a total of 107 patients with Type 1 diabetes in poor glycaemic control (HbA(1c) > or = 8%) were assigned to a 6-week CBGT or blood glucose awareness training (BGAT) as control condition. The intervention was preceded by a 3-month run-in period. Glycaemic control (HbA(1c)), diabetes-specific self efficacy (CIDS), diabetes-related distress (PAID) and depressive symptoms (CES D), were assessed at baseline (T1), directly before (T2) and 3 months after (T3) the intervention. RESULTS: No significant changes in HbA(1c) were found after CBGT, whilst diabetes self-efficacy increased (mean CIDS score 71.6 +/- 14.0 to 74.3 +/- 12.2) and diabetes-related distress (mean PAID score 47.0 +/- 21.6 to 42.6 +/- 20.8) and depressive symptoms decreased (mean CES-D score 16.9 +/- 12.8 to 13.5 +/- 12.6). Changes in psychological outcomes were similar for both treatment groups. Diabetes self-care behaviours improved equally. Drop-out rate, which was higher among CBGT participants, was relatively low overall (total n = 15, 17.05%), and both interventions were well-appreciated by the participants. CONCLUSIONS: CBGT was successful in improving self-efficacy, diabetes-related distress and mood at 3 months' follow-up, but not in improving glycaemic control. PMID- 16241933 TI - Creatine kinase elevation in a patient taking rosiglitazone. PMID- 16241934 TI - Incidence and outcome of asymptomatic bacteriuria in females with Type 2 diabetes mellitus over a 1-year follow-up period and association with risk factors. PMID- 16241935 TI - Screening for occult cancer in patients with acute venous thromboembolism. PMID- 16241936 TI - Decision analysis for cancer screening in idiopathic venous thromboembolism. AB - BACKGROUND: The SOMIT trial randomized patients with idiopathic venous thromboembolism (IVTE) and without signs of cancer at routine medical examination, to extensive screening for cancer plus 2 years of follow-up or to just 2-year follow-up. METHODS: The data of the SOMIT-trial were used to perform a decision analysis. The screening tests were divided in several possible strategies. The number of detected cancer patients and the number of patients investigated further for an eventually benign condition were calculated for each strategy. The total costs for the screening strategy and for each detected cancer patient were determined. Based on the tumor type, stage, age and gender of the individual cancer patient, the difference in live years gained (LYG) was calculated between the two study groups. RESULTS: Computed tomography (CT) of the abdomen combined with sputum cytology and mammography detected 12 of the 14 patients with cancer and had one false-positive result. In general, screening strategies including abdominal/pelvic ultrasonography (US) or tumor markers yielded a higher number of patients needed to screen in comparison with those using abdominal/pelvic CT. Furthermore, the strategies which included colonoscopy, tumor markers, and abdominal/pelvic US were significantly more costly, had inferior LYG and higher costs per LYG, when compared with strategies using abdominal/pelvic CT. CONCLUSIONS: Despite the limitations of this analysis, the screening for cancer with a strategy including abdominal/pelvic CT with or without mammography and/or sputum cytology appears potentially useful for cancer screening in patients with IVTE. The cost-effectiveness analysis of this strategy needs confirmation in a large trial. PMID- 16241937 TI - Mathematical and biological models of blood coagulation. PMID- 16241938 TI - During coagulation, thrombin generation shifts from chemical to diffusional control. PMID- 16241939 TI - Determinants of specificity in coagulation proteases. AB - Proteases play diverse roles in a variety of essential biological processes, both as non-specific catalysts of protein degradation and as highly specific agents that control physiologic events. Here, we review the mechanisms of substrate specificity employed by serine proteases and focus our discussion on coagulation proteases. We dissect the interplay between active site and exosite specificity and how substrate recognition is regulated allosterically by Na+ binding. We also draw attention to a functional polarity that exists in the serine protease fold, which sheds light on the structural linkages between the active site and exosites. PMID- 16241940 TI - Lepirudin in patients with heparin-induced thrombocytopenia - results of the third prospective study (HAT-3) and a combined analysis of HAT-1, HAT-2, and HAT 3. AB - OBJECTIVES: To assess efficacy and safety of lepirudin in patients with heparin induced thrombocytopenia (HIT) in a prospective study (HAT-3) as well as in a combined analysis of all HAT study data. PATIENTS/METHODS: Patients with laboratory-confirmed HIT were treated with lepirudin in three different aPTT adjusted dose regimen and during cardiopulmonary bypass (CPB). Endpoints were new thromboembolic complications (TEC), limb amputations, and death and major bleeding. A historical control group (n = 120) was used for comparison. RESULTS: After start of lepirudin in 205 patients treated in HAT-3, the combined endpoint occurred in 43 (21.0%). Thirty (14.6%) patients died, 10 (4.9%) underwent limb amputation, and 11 (5.4%) new TECs occurred. Major bleeding occurred in 40 patients (19.5%) (seven during CPB surgery). Combining all prospective HAT trials (n = 403), after start of lepirudin treatment, the combined endpoint occurred in 82 patients (20.3%), with 47 deaths (11.7%), 22 limb amputations (5.5%), 30 new TECs (7.4%), and 71 (17.6%) major bleedings. Compared with the historical control group (log-rank test), the combined endpoint after start of treatment was reduced (29.7% vs. 52.1%, P = 0.0473), primarily because of reduction in new thromboses (11.9% vs. 32.1%, P = 0.0008). Mean lepirudin maintenance doses ranged from 0.07 to 0.11 mg kg(-1) h(-1). Major bleeding was more frequent in the lepirudin treated patients (29.4% vs. 9.1%, P = 0.0148). CONCLUSIONS: The rate of new TECs in HIT patients is low after start of lepirudin treatment. The rate of major bleeding of 17.6% might be reduced by reducing the starting dose to 0.1 mg kg(-1) h(-1). PMID- 16241941 TI - Attitudes toward genetic testing for thrombophilia in asymptomatic members of a large family with heritable protein C deficiency. AB - BACKGROUND: Little research has been performed regarding the psychological consequences of knowing that one is at an increased risk for venous thrombosis. OBJECTIVES: The aim of this study was to explore attitudes toward genetic testing for protein C deficiency. METHODS: Questionnaires about genetic testing attitudes, dispositional anxiety, risk perception, and thrombosis-related worry were completed by 168 asymptomatic members of a North-American kindred with a high incidence of heritable protein C deficiency conferring a high lifetime risk of venous thrombosis. A total of 76 subjects (45%) had not been tested for protein C deficiency before participating in our study whereas the other 92 subjects (55%) had been tested prior to filling in the questionnaire, of whom 34 people had protein C deficiency, while 58 did not. RESULTS: Family members with protein C deficiency perceived a higher risk of suffering venous thrombosis and scored higher on thrombosis-related worry than family members without protein C deficiency. Participants who had not been tested did not report excessive thrombosis-related worry. Participants with protein C deficiency reported a belief in the psychological and health benefits of testing, and felt that they experienced low psychological distress following the genetic test. High psychological distress following the test was related to dispositional anxiety and thrombosis-related worry. Participants without protein C deficiency were relieved after finding out that they did not have the deficiency. CONCLUSION: There seem to be few negative psychological consequences of knowing that one is at an increased risk for venous thrombosis, except in vulnerable individuals. PMID- 16241942 TI - Determinants of plasma D-dimer levels in a traveling population. AB - BACKGROUND: Plasma D-dimer measurement is a widely used diagnostic test for assessing individuals with suspected venous thromboembolism (VTE). Whilst a negative test is helpful in ruling out thrombosis, the significance and determinants of an elevated plasma D-dimer level in otherwise healthy subjects are poorly understood. OBJECTIVES: To determine the association between recognized risk factors for VTE and plasma D-dimer levels in an adult population. SUBJECTS AND METHODS: Blood samples for measurement of plasma D-dimer levels were obtained from 1000 adults aged <70 years who were participating in a study investigating the incidence of VTE in long distance air travellers. The relationship between D-dimer levels and selected risks factors for VTE including thrombophilia status was investigated. RESULTS: The median (Inter-quartile range) D-dimer level was 243 ng mL(-1) (175-345). Multivariate analysis showed that plasma D-dimer levels were positively associated with increasing age, larger body mass index, female gender, the use of hormone therapy, thrombophilia state, and the presence of co-morbid conditions. CONCLUSION: Plasma D-dimer levels vary markedly between individuals and are associated with known risk factors for VTE, including the presence of thrombophilia conditions. The potential role for the measurement of plasma D-dimer as a marker for thrombosis risk requires further investigation. PMID- 16241943 TI - Helical computed tomography and alternative diagnosis in patients with excluded pulmonary embolism. AB - OBJECTIVE: A clinical diagnosis of pulmonary embolism (PE) is confirmed objectively in 20-30% of patients. Helical computed tomography (CT) can allow an alternative diagnosis to be made. The frequency and validity of alternative diagnoses on helical CT in consecutive patients presenting with clinically suspected PE was assessed. PATIENTS AND METHODS: In all 512 prospectively analyzed patients helical CT scan was performed, and apart from presence or absence of PE, pathologic changes in lung parenchyma, mediastinum, cardiovascular system, pleura and skeleton were recorded. When possible an alternative diagnosis was given and compared with the final diagnosis after 3 months follow-up. RESULTS: In 130 patients (25.4%) PE was excluded and an alternative diagnosis considered likely. In 123 of the 130 patients (94.6%) this diagnosis was unchanged at 3 months follow-up. The diagnoses included pneumonia (n = 67), malignancy (n = 22), pleural fluid (n = 10), cardiac failure (n = 10), COPD (n = 6) and a variety of other causes (n = 15). The diagnosis changed at follow-up in seven patients (5.4%). An initial diagnosis of pneumonia changed to malignancy in two patients and to pleuritis and cardiac failure in one patient each. In two other patients malignancy and chronic obstructive pulmonary disease (COPD) were ruled out and the diagnosis changed to pneumonia. In one patient the final diagnosis remained unknown after an initial suspicion of malignancy. CONCLUSION: In clinically suspected PE helical CT allows a reliable alternative diagnosis to be made in 25.4% of patients. This feature is an unique advantage in comparison with other diagnostic tests and supports the decision of taking helical CT as first line test in suspected PE. PMID- 16241944 TI - Differential value of risk factors and clinical signs for diagnosing pulmonary embolism according to age. AB - INTRODUCTION: The diagnostic value of clinical presentation of pulmonary embolism (PE) is uncertain in the elderly, who often have concomitant cardiopulmonary diseases that may mimic PE. The aim of our study was to assess the differential value of risk factors, symptoms and clinical signs of venous thromboembolism, results of electrocardiogram and chest X-ray for the diagnosis of PE in suspected patients according to age. METHODS: We analyzed data from two outcome studies which enrolled 1721 consecutive patients presenting in the emergency department with clinically suspected PE defined as acute onset of new or worsening shortness of breath or chest pain without any other obvious etiology. All patients underwent a sequential diagnostic work-up and a 3-month follow-up. RESULTS: The proportion of confirmed PE was 24.2% (416 of 1721). Strength of the association with PE did not differ according to age group for history of venous thromboembolism (VTE), recent surgery, tachypnea at admission or right ventricular strain on electrocardiogram. Active malignancy, hemoptysis, tachycardia, hemidiaphragmatic elevation and pleural effusion at chest X-ray were no more associated with PE in the patients aged of 75 years or more. Finally, symptoms and signs of deep venous thrombosis, and an alternative diagnosis less probable than PE were associated with PE in all age groups, but the strength of this association decreased significantly with advancing age. CONCLUSION: Some risk factors, symptoms and signs of VTE are less strongly or even not at all associated with PE in the elderly. Physicians should take this into account when attending elderly patients suspected of PE and when assessing their clinical probability of PE. PMID- 16241945 TI - Old and new risk factors for upper extremity deep venous thrombosis. AB - BACKGROUND: Well known risk factors for upper extremity deep venous thrombosis are the presence of a central venous catheter (CVC) and malignancy, but other potential risk factors, such as surgery, injury and hormone replacement therapy (HRT), have not yet been explored. METHODS: We performed a population-based case control study including 179 consecutive patients, aged 18-70 years with upper extremity deep venous thrombosis and 2399 control subjects. Participants reported on acquired risk factors in a questionnaire and factor V Leiden and prothrombin 20210A mutation were ascertained. Information on CVC was obtained from discharge letters. RESULTS: Forty-two patients (23%) and one control subject (0.04%) had a CVC (ORadj: 1136, 95% CI: 153-8448, adjusted for age and sex). Cancer patients without a CVC had an eightfold increased risk of venous thrombosis of the arm (ORcrude: 7.7, 95% CI: 4.6-13.0). Other evident risk factors were prothrombotic mutations, surgery, immobilization of the arm (plaster cast), oral contraceptive use and family history, with odds ratios varying from 2.0 up to 13.1. The risk in the presence of injury and during puerperium was twofold or more increased, although not significantly. In contrast HRT, unusual exercise, travel and obesity did not increase the risk. Hormone users had an increased risk in the presence of prothrombotic mutations or surgery. Obese persons (BMI > 30 kg m(-2)) undergoing surgery had a 23-fold increased risk of arm thrombosis compared with non-obese persons not undergoing surgery. CONCLUSION: A CVC is a very strong risk factor for arm thrombosis. Most risk factors for thrombosis in the leg are also risk factors for arm thrombosis. PMID- 16241946 TI - BAY 59-7939: an oral, direct factor Xa inhibitor for the prevention of venous thromboembolism in patients after total knee replacement. A phase II dose-ranging study. AB - BACKGROUND: BAY 59-7939, a novel, oral, direct factor Xa inhibitor, is in clinical development for the prevention of venous thromboembolism (VTE), a frequent complication following orthopaedic surgery. METHODS: In a multicenter, parallel-group, double-blind, double-dummy study, 621 patients undergoing elective total knee replacement were randomly assigned to oral BAY 59-7939 (2.5, 5, 10, 20, and 30 mg b.i.d., initiated 6-8 h postsurgery), or subcutaneous enoxaparin (30 mg b.i.d., initiated 12-24 h postsurgery). Treatment was continued until mandatory bilateral venography 5-9 days after surgery. The primary efficacy endpoint was a composite of any deep vein thrombosis (proximal and/or distal), confirmed non-fatal pulmonary embolism and all-cause mortality during treatment. The primary safety endpoint was major, postoperative bleeding during treatment. RESULTS: Of the 613 patients treated, 366 (59.7%) were evaluable for the primary efficacy analysis. The primary efficacy endpoint occurred in 31.7%, 40.4%, 23.3%, 35.1%, and 25.4% of patients receiving 2.5, 5, 10, 20 and 30 mg b.i.d. doses of BAY 59-7939, respectively (test for trend, P = 0.29), compared with 44.3% in the enoxaparin group. The frequency of major, postoperative bleeding increased with increasing doses of BAY 59-7939 (test for trend, P = 0.0007), with no significant difference between any dose group compared with enoxaparin. Bleeding endpoints were lower for the 2.5-10 mg b.i.d. doses compared with higher doses of BAY 59 7939. CONCLUSIONS: Oral administration of 2.5-10 mg b.i.d. of BAY 59-7939, early in the postoperative period, showed potential efficacy and an acceptable safety profile, similar to enoxaparin, for the prevention of VTE in patients undergoing elective total knee replacement. PMID- 16241947 TI - A novel polymorphism in the factor XIII B-subunit (His95Arg): relationship to subunit dissociation and venous thrombosis. AB - BACKGROUND: Factor (F)XIII B-subunit, which plays a carrier role for zymogen FXIIIA, is highly polymorphic, but the molecular basis for these polymorphisms and their relationship to disease remains unknown. OBJECTIVES: To screen the FXIIIB gene coding region for common variation and analyze possible functional effects. METHODS AND RESULTS: We examined the FXIIIB gene by PCR-SSCP and identified three common single nucleotide polymorphisms: A8259G, C29470T and A30899G. A8259G results in substitution of His95Arg in the second Sushi domain. An FXIII tetramer ELISA was developed to analyze B-subunit dissociation from A subunit (leading to access to the catalytic site of FXIII). Increased subunit dissociation, 0.51 vs. 0.45 (fraction of total tetramer), was found in plasma from subjects possessing the Arg-allele. However, when the variants were purified to homogeneity and binding was analyzed by steady-state kinetics, no difference was observed. The relationship between His95Arg and venous thrombosis was investigated in 214 patients and 291 controls from Leeds. His/Arg + Arg/Arg genotypes were more frequent in patients than controls (22.4% vs. 15.1%). His95Arg was also investigated in the Leiden Thrombophilia Study, in which a similar difference was observed for 471 patients vs. 472 controls (18.5% vs. 14.0%), for a pooled odds ratio (OR) of 1.5 (CI95 1.1-2.0). CONCLUSIONS: We have identified three FXIIIB polymorphisms, one of which codes for substitution of His95Arg. The Arg95 variant associates with a moderately increased risk for venous thrombosis, and with increased dissociation of the FXIII subunits in plasma, although in vitro steady-state binding between purified subunits was not affected. PMID- 16241948 TI - Thrombin generation profiles in deep venous thrombosis. AB - BACKGROUND: Reliable markers and methods to predict risk for thrombosis are essential to clinical management. OBJECTIVE: Using an integrated approach that defines an individual's comprehensive coagulation phenotype might prove valuable in identifying individuals at risk for experiencing a thrombotic event. METHODS: Using a numerical simulation model, we generated tissue factor (TF) initiated thrombin curves using coagulation factor levels from the Leiden Thrombophilia Study population and evaluated thrombotic risk, by sex, age, smoking, alcohol consumption, body mass index (BMI) and oral contraceptive (OC) use. We quantitated the initiation, propagation and termination phases of each individuals' comprehensive TF-initiated thrombin generation curve by the parameters: time to 10 nm of thrombin, maximum time, level and rate (MaxR) of thrombin generated and total thrombin. RESULTS: The greatest risk association was obtained using MaxR; with a 2.6-fold increased risk at MaxR exceeding the 90th percentile. The odds ratio (OR) for MaxR was 3.9 in men, 2.1 in women, and 2.9 in women on OCs. The association of risk with thrombin generation did not differ by age (OR:2.8 OR:2.5), BMI (OR:2.9 OR:2.3) or alcohol use. In both numerical simulations and empirical synthetic plasma, OC use created extreme shifts in thrombin generation in both control women and women with a prior thrombosis, with a larger shift in thrombin generation in control women. This suggests an interaction of OC use with underlying prothrombotic abnormalities. CONCLUSIONS: Thrombin generation based upon the individual's blood composition is associated with the risk for thrombosis and may be useful as a predictive marker for evaluating thrombosis on an individual basis. PMID- 16241949 TI - Urokinase induced fibrinolysis in thromboelastography: a model for studying fibrinolysis and coagulation in whole blood. AB - BACKGROUND: The contact system (CS) proteins, factor XII and prekallikrein are thought to have roles in blood coagulation and fibrinolysis. Recent research has suggested that the CS proteins might be more important in fibrinolysis and cell function than in coagulation. Most studies on fibrinolysis have used plasma or euglobulin assays, ignoring the influence of cellular elements of blood on the fibrinolytic process. OBJECTIVE AND METHODS: In order to study both coagulation and fibrinolysis in whole blood (WB), we have developed a thromboelastography (TEG) assay to investigate both coagulation and fibrinolysis in the same blood sample. In this assay, named urokinase (UK) induced fibrinolysis in thromboelastography (UKIFTEG), TEG is performed on recalcified citrated WB in the presence of UK. Large variations in Ly60 (percentage lysis 60 min after clot formation) were obtained between different donors with the same UK concentration. The UKIFTEG assay was therefore performed using UK concentrations that gave Ly60 values in the approximate range of 20-40%. RESULTS: The effect of CS activation was investigated in the presence or absence of celite (10 mg mL(-1) blood). Celite shortened the clotting time (CT), and increased Ly60 values. Factor XIIa (FXIIa) and plasma kallikrein (KK) produced concentration dependent reductions in CT (significant at concentrations of 1303 and 2600 ng mL(-1) blood, respectively) and increased Ly60 values (significant at concentrations of 652 and 1300 ng mL( 1) blood, respectively). CONCLUSIONS: Our results show that CS activation and both FXIIa and KK produce reductions in clotting time and enhanced fibrinolysis in UKIFTEG. PMID- 16241950 TI - Adhesive surface determines raft composition in platelets adhered under flow. AB - Adhesion to von Willebrand factor (VWF) induces platelet spreading, whereas adhesion to collagen induces aggregation. Here we report that cholesterol-rich domains (CRDs) or rafts play a critical role in clustering of receptors that control these responses. Platelets adhered to VWF and collagen show CRDs concentrated in filopodia which contain both the VWF receptor glycoprotein (GP) Ibalpha and the collagen receptor GPVI. Biochemical analysis of CRDs shows a threefold enrichment of GPIbalpha (but not GPVI) in VWF-adhered platelets and a fourfold enrichment of GPVI (but not GPIbalpha) in collagen-adhered platelets. Depletion of cholesterol (i) leaves the initial adhesion unchanged, (ii) inhibits spreading on VWF and aggregate formation on collagen, (iii) leaves filopodia formation intact, and (iv) reduces the localization in filopodia of GPIbalpha but not of GPVI. These data show that the adhesive substrate determines the composition of CRDs, and that cholesterol is crucial for redistribution of GPIbalpha but not of GPVI. PMID- 16241951 TI - Intraplatelet signaling mechanisms of the priming effect of matrix metalloproteinase-2 on platelet aggregation. AB - OBJECTIVE: Platelets contain and release some matrix metalloproteinases (MMPs), enzymes involved in the degradation of extracellular matrix, and one of these (MMP-2) exerts a proaggregatory effect. We explored the signal transduction mechanisms activated by MMP-2 in human blood platelets. METHODS AND RESULTS: Recombinant, human MMP-2, added before stimulation with subthreshold doses of different agonists, potentiated platelet activation, calcium influx, IP3 formation, and pleckstrin phosphorylation. Wortmannin and LY29400, two PI3-K inhibitors, suppressed the potentiating effects of MMP-2 and preincubation with MMP-2 enhanced the thrombin-induced association of the p85alpha PI3-K subunit with the cytoskeleton and increased the phosphorylation of PKB. Protein tyrosine kinase inhibitors, MAP kinase inhibitors, PLA2 inhibitors, cyclooxygenase inhibitors and antagonists of the P2Y1 and P2Y12 receptors did not affect the potentiating activity of MMP-2 on platelets. CONCLUSION: Our data show that MMP 2, at a concentration released by activated platelets, facilitates platelet activation acting at the level of a second messenger system common to different agonists and related to the activation of PI3-K. Platelet-released MMP-2 may contribute to platelet activation in vivo. PMID- 16241952 TI - Two subpopulations of thrombin-activated platelets differ in their binding of the components of the intrinsic factor X-activating complex. AB - Binding of fluorescein-labeled coagulation factors IXa, VIII, X, and allophycocyanin-labeled annexin V to thrombin-activated platelets was studied using flow cytometry. Upon activation, two platelet subpopulations were detected, which differed by 1-2 orders of magnitude in the binding of the coagulation factors and by 2-3 orders of magnitude in the binding of annexin V. The percentage of the high-binding platelets increased dose dependently of thrombin concentration. At 100 nm of thrombin, platelets with elevated binding capability constituted approximately 4% of total platelets and were responsible for the binding of approximately 50% of the total bound factor. Binding of factors to the high-binding subpopulation was calcium-dependent and specific as evidenced by experiments in the presence of excess unlabeled factor. The percentage of the high-binding platelets was not affected by echistatin, a potent aggregation inhibitor, confirming that the high-binding platelets were not platelet aggregates. Despite the difference in the coagulation factors binding, the subpopulations were indistinguishable by the expression of general platelet marker CD42b and activation markers PAC1 (an epitope of glycoprotein IIb/IIIa) and CD62P (P-selectin). Dual-labeling binding studies involving coagulation factors (IXa, VIII, or X) and annexin V demonstrated that the high-binding platelet subpopulation was identical for all coagulation factors and for annexin V. The high-binding subpopulation had lower mean forward and side scatters compared with the low-binding subpopulation ( approximately 80% and approximately 60%, respectively). In its turn, the high-binding subpopulation was not homogeneous and included two subpopulations with different scatter values. We conclude that activation by thrombin induces the formation of two distinct subpopulations of platelets different in their binding of the components of the intrinsic fX-activating complex, which may have certain physiological or pathological significance. PMID- 16241953 TI - A novel nitric oxide-releasing statin derivative exerts an antiplatelet/antithrombotic activity and inhibits tissue factor expression. AB - BACKGROUND: NO-releasing statins are new chemical entities, combining HMG-CoA reductase inhibition and slow NO release, that possess stronger anti-inflammatory and antiproliferative activities than the native statins. OBJECTIVE: We evaluated the antithrombotic effects of nitropravastatin (NCX-6550) by assessing its activity on platelet activation and tissue factor (TF) expression by mononuclear cells in vitro and in vivo. METHODS AND RESULTS: In vitro, NCX-6550 inhibited (1) U46619- and collagen-induced platelet aggregation in buffer and plasma; (2) collagen-induced P-selectin expression in whole blood and (3) platelet adhesion to collagen-coated coverslips under high shear stress. These effects were displayed at concentrations of NCX-6550 ranging from 25 to 100 mum, and were totally reverted by the guanylylcyclase inhibitor ODQ (10 microm). Equimolar concentrations of pravastatin had no influence on these parameters of platelet function. LPS- and PMA-induced TF expression by blood mononuclear cells was also inhibited by NCX-6550 (IC50 13 microm), but not by pravastatin, as assessed by functional and immunological assays and by real-time PCR. In a mouse model of platelet pulmonary thromboembolism, induced by the i.v. injection of collagen plus epinephrine, pretreatment with NCX-6550 (24-48 mg kg(-1)) significantly reduced platelet consumption, lung vessel occlusion and mortality. Moreover, nitropravastatin markedly inhibited the generation of procoagulant activity by spleen mononuclear cells and peritoneal macrophages in mice treated with LPS. In these in vivo models too, pravastatin failed to affect platelet activation and monocyte/macrophage procoagulant activity. CONCLUSIONS: Our results show that nitropravastatin exerts strong antithrombotic effects in vitro and in vivo, and may represent an interesting antiatherothrombotic agent for testing in acute coronary syndromes. PMID- 16241954 TI - Effects of platelet binding on whole blood flow cytometry assays of monocyte and neutrophil procoagulant activity. AB - BACKGROUND: Monocytes and neutrophils form heterotypic aggregates with platelets initially via engagement of platelet surface P-selectin with leukocyte surface P selectin glycoprotein ligand-1 (PSGL-1). The resultant intracellular signaling causes the leukocyte surface expression of tissue factor and activation of leukocyte surface Mac-1 (integrin alphaMbeta2, CD11b/CD18). The activation dependent conformational change in monocyte surface Mac-1 results in the binding of coagulation factor Xa (FXa) and/or fibrinogen to Mac-1. The aim of this study was to develop whole blood flow cytometry assays of these procoagulant activities and to investigate the effects of platelet binding to monocytes and neutrophils. METHODS: Citrate or D-Phe-Pro-Arg-chloromethylketone (PPACK) anticoagulated whole blood was incubated with monoclonal antibodies against CD14 (PECy5), CD42a (PE), FITC-conjugated test antibody and an agonist, and then fixed with FACS lyse. Appropriate isotype negative controls were prepared in parallel. A BD FACSCalibur was used to analyze monocytes and neutrophils, which were identified based on CD14 fluorescence, forward and 90 degrees light scatter. These populations were further gated into CD42a-positive (platelet-bound) and CD42a-negative (platelet free). Geometric mean fluorescence and per cent positive data were collected for each subpopulation to measure the binding of test antibodies directed at CD42a, tissue factor, coagulation FXa, bound fibrinogen, activated Mac-1, and CD11b. Compensation controls were prepared on six normal donors prior to the study and these settings were used throughout the 10 donor study. Negative controls verified the lack of cross talk, particularly in the quantified FITC and PE parameters. RESULTS: The physiologic agonists collagen and ADP increased monocyte platelet and neutrophil-platelet aggregates and increased leukocyte surface Mac 1/CD11b and surface-bound tissue factor, FXa and fibrinogen. Whereas the increases in Mac-1/CD11b were mainly independent of leukocyte-platelet binding, the increases in surface-bound tissue factor, FXa and fibrinogen were mainly dependent on leukocyte-platelet binding. CONCLUSIONS: (i) We have developed novel whole blood flow cytometry assays to measure bound tissue factor, coagulation FXa, fibrinogen, activated Mac-1 and CD11b on the surface of monocytes and neutrophils, allowing independent analysis of monocytes and neutrophils with and without surface-adherent platelets. (ii) The monocyte and neutrophil surface binding of tissue factor, FXa and fibrinogen is mainly dependent on platelet adherence to monocytes and neutrophils, whereas the monocyte and neutrophil surface expression of CD11b and activated Mac-1 is mainly independent of platelet adherence to monocytes and neutrophils. PMID- 16241955 TI - Classification of venous thromboembolism (VTE). The clot is hot: inflammation, myeloid leukocytes, and venous thromboembolism. PMID- 16241956 TI - Classification of venous thromboembolism (VTE). Role of acute inflammatory stress in venous thromboembolism. PMID- 16241957 TI - Classification of venous thromboembolism (VTE). Platelets in venous thrombosis. PMID- 16241958 TI - APC-PCI complex concentration is higher in patients with previous venous thromboembolism with Factor V Leiden. PMID- 16241959 TI - Harris syndrome - a geographic perspective. PMID- 16241960 TI - Role of angiotensin-converting enzyme insertion/deletion and plasminogen activator inhibitor-1 4G/5G gene polymorphisms in retinal vein occlusion. PMID- 16241961 TI - Protein Z in pregnancy: exaggerated rise in obese women. PMID- 16241962 TI - Antiphospholipid antibodies and antibodies to tissue factor pathway inhibitor in women with implantation failures or early and late pregnancy losses. PMID- 16241963 TI - Rituximab-induced long-term remission in patients with refractory acquired hemophilia. PMID- 16241964 TI - The DNA-pooling technique allowed for the identification of three novel mutations responsible for afibrinogenemia. PMID- 16241965 TI - Lepirudin: is the approved dosing schedule too high? PMID- 16241966 TI - More on: is there a need for replacement of the international reference preparation for thromboplastin, bovine, combined (OBT/79)? PMID- 16241967 TI - Polymorphism and hemophilia A causing inversions in distal Xq28: a complex picture. PMID- 16241968 TI - More on: polymorphism and hemophilia A causing inversions in distal Xq28: a complex picture. PMID- 16241970 TI - Guidelines on use of anti-IFN-beta antibody measurements in multiple sclerosis: report of an EFNS Task Force on IFN-beta antibodies in multiple sclerosis. AB - Therapy-induced binding and neutralizing antibodies is a major problem in interferon (IFN)-beta treatment of multiple sclerosis. The objective of this study was to provide guidelines outlining the methods and clinical use of the measurements of binding and neutralizing antibodies. Systematic search of the Medline database for available publications on binding and neutralizing antibodies was undertaken. Appropriate publications were reviewed by one or more of the task force members. Grading of evidence and recommendations was based on consensus by all task force members. Measurements of binding antibodies are recommended for IFN-beta antibody screening before performing a neutralizing antibody (NAB) assay (Level A recommendation). Measurement of NABs should be performed in specialized laboratories with a validated cytopathic effect assay or MxA production assay using serial dilution of the test sera. The NAB titre should be calculated using the Kawade formula (Level A recommendation). Tests for the presence of NABs should be performed in all patients at 12 and 24 months of therapy (Level A recommendation). In patients who remain NAB-negative during this period measurements of NABs can be discontinued (Level B recommendation). In patient with NABs, measurements should be repeated, and therapy with IFN-beta should be discontinued in patients with high titres of NABs sustained at repeated measurements with 3- to 6-month intervals (Level A recommendation). PMID- 16241971 TI - Human photosensitivity: from pathophysiology to treatment. AB - Photosensitivity is a condition detected on the electroencephalography (EEG) as a paroxysmal reaction to Intermittent Photic Stimulation (IPS). This EEG response, elicited by IPS or by other visual stimuli of daily life, is called Photo Paroxysmal Response (PPR). PPRs are well documented in epileptic and non epileptic subjects. Photosensitivity rarely in normal individuals evolves into epilepsy. Photosensitive epilepsy is a rare refex epilepsy characterized by seizures in photosensitive individuals. The development of modern technology has increased the exposition to potential seizure precipitants in people of all ages, but especially in children and adolescents. Actually, videogames, computers and televisions are the most common triggers in daily life of susceptible persons. The mechanisms of generation of PPR are poorly understood, but genetic factors play an important rule. The control of visually induced seizures has, generally a good prognosis. In patients known to be visually sensitive, avoidance of obvious source and stimulus modifications are very important and useful to seizure prevention, but in the large majority of patients with epilepsy and photosensitivity antiepileptic drugs are needed. PMID- 16241972 TI - Relationship between nigrostriatal dopaminergic degeneration, urinary symptoms, and bladder control in Parkinson's disease. AB - Patients with Parkinson's disease (PD) often have lower urinary tract symptoms (LUTS). Studies have indicated a correlation between dopaminergic degeneration and LUTS and presence of overactive bladder. We evaluated 18 patients with Parkinson's disease using single-photon emission computerized tomography (SPECT) imaging of the dopamine transporter with [(123)I]-FP-CIT, and bladder symptoms were assessed using questionnaires and full urodynamic evaluation both in medicated state and after cessation. Bladder symptoms correlated with age, stage and severity of disease but not with uptake of the ligand in the striatum. Patients with bladder symptoms had a significant lower uptake in the striatum compared with patients without LUTS. In patients with severe bladder dysfunction, LUTS correlated with putamen/caudate ratio. The specific binding of the ligand did not correlate with urodynamics parameters or any change in these after wash out. Our findings suggest that the presence of LUTS is associated with the degeneration of the total number of nigrostriatal dopaminergic neurones, whilst the severity of bladder dysfunction is correlated with the relative degeneration of the caudate nucleus. The effects of medication on bladder control, as evaluated by urodynamics are believed to involve structures outside the basal ganglia. PMID- 16241973 TI - Absence of spinocerebellar ataxia type 3/Machado-Joseph disease within ataxic patients in the Czech population. AB - Although spinocerebellar ataxia type 3 (SCA3)/Machado-Joseph disease is the most common type of SCA worldwide, we did not identify any cases of the disease amongst SCA patients in the Czech population. It has been proposed that the prevalence of large normal alleles correlates with the frequency of various types of SCA. We have therefore attempted to resolve the absence of SCA3 in our population by investigating, within 204 normal chromosomes, the frequency and nature of CAG repeats as well as two intragenic polymorphisms. We found that large normal alleles with more than 33 CAG repeats were observed at a frequency of only 0.49%. Whereas most of the expanded alleles worldwide have the CA haplotype, this was the least common (5.4%) variant observed in our study, although it was associated with a larger mean CAG repeat length (26.9). We postulate that the absence of SCA3 in the Czech population might be explained by the lack of large normal alleles and consequently a relatively small reservoir for aberrant CAG expansions at the SCA3 locus. PMID- 16241974 TI - The impact of paclitaxel or cisplatin-based chemotherapy on sympathetic skin response: a prospective study. AB - The current study aimed to assess the viability of sympathetic sudomotor fibers in cancer patients treated with cisplatin or paclitaxel-based chemotherapy and to ascertain whether this method could contribute to the diagnostic sensitivity of conventional techniques. Sympathetic skin response (SSR) from the hand and sole of 23 cancer patients (nine females and 14 males, mean age 62.4 +/- 10.5 years) was recorded unilaterally before and after chemotherapy with six courses of cumulative cisplatin or paclitaxel containing regimens. Clinical and electrophysiological data were also collected and correlated with the SSR results. Twenty-three healthy subjects served as controls. SSR abnormalities were only present in patients with evidence of peripheral neuropathy assessed by conventional nerve conduction techniques. Three patients had absent SSR in the upper limb whilst six patients had absent SSR both in the upper and lower limbs. In the upper limb, the mean SSR latency was not significantly altered through time (P = 0.086). In the lower limb the mean delay from baseline to follow-up was significantly changed (P = 0.029). In patients, the mean SSR latency was significantly prolonged compared with controls in both upper limb (P = 0.001) and lower limb (P = 0.000). SSR abnormalities were strongly related to sensory conduction abnormalities as detected by conventional techniques (r = 0.39, P = 0.004). Our results showed that SSR does not seem to add to the diagnostic sensitivity of conventional techniques in chemotherapy-induced neuropathy. However, its role in the disclosure of small fibers neuropathy abnormalities is worth considering. Further studies are warranted to address this important issue. PMID- 16241975 TI - Medical resource use and costs of health care after acute stroke in Germany. AB - The purpose of this study was to determine the 12 months medical resource use following admission to hospital with acute stroke and to calculate costs from a societal perspective. Data of consecutive patients with confirmed stroke were analysed. Acute hospital data were taken from medical records, socio-demographic variables from patients' interviews. A follow-up questionnaire about resource utilization was completed by patients or proxies 12 months after acute hospital admission. Costs were calculated by multiplying medical resource units used with cost factors per unit. Mean age of a total of 383 patients was 65 years and 41% were female. The median length of the initial stay in the acute hospital was 12 days at an average cost of 4650 per patient (49% of direct costs). Rehabilitation (16%), readmission (11%), medication (9%), and nursing costs (6%) were other contributors to the direct costs which amounted to a total of 9452 +/- 7599 per patient during 12 months. Indirect cost amounted to a total of 2014 +/- 5312. Patients' age, severity and type of stroke influenced the total stroke-associated costs. The large economic burden of stroke indicates the need for assessing and improving efficient health care for affected patients. PMID- 16241976 TI - Clinical and immune responses correlate in glatiramer acetate therapy of multiple sclerosis. AB - Glatiramer acetate (GA) treatment for relapsing remitting multiple sclerosis (RRMS) leads to decreased GA-specific proliferative responses and a Th2 cytokine shift. To study a possible correlation between immunological and clinical responses to GA therapy, we prospectively followed RRMS patients clinically, by magnetic resonance imaging and by primary immunological assays. Fluctuation of GA specific proliferative responses was significantly lower in treatment responders than in untreated patients, and GA-specific proliferative responses were increased during relapses. These associations suggest a possible causal relationship between immunological and clinical responses to GA therapy. Primary proliferation assays may thus be a useful marker for treatment response. PMID- 16241977 TI - Accidental MCI in healthy subjects: a prospective longitudinal study. AB - A study was realized on 130 healthy and autonomous volunteers (60-80 years old) who met specific medical and functional inclusion criteria. A comprehensive battery of neuropsychological tests was performed at baseline (M0), 6 and 12 months (M6, M12). At M0 the results indicated that 65% were cognitively normal on each of all the neuropsychological tests, whereas 35% presented a cognitive deficit on one or more tests. At M12, 52% of the subjects who had a cognitive deficit at M0 remained impaired, whereas 48% normalized their scores: they performed as well as the subjects classified normal at M0. The results also indicated that the subjects who remained impaired at M12, had at M0 low scores on three tests or more, whereas the ones who normalized their scores had one or two failed tests. This study focuses on the risk of false positive cases and shows that low scores can be accidental. The authors propose decision rules allowing to reduce the risk of false positive cases. The observation of accidental impairment invites to be cautious and makes this 1-year follow-up study particularly relevant, since a 1-year follow-up is generally needed to diagnose very mild dementia. PMID- 16241978 TI - Lateralized ictal immobility of the upper limb in patients with temporal lobe epilepsy. AB - The primary aim of this study was to establish the incidence and the lateralizing value of 'lateralized ictal immobility of the upper limb' (LIL) in patients suffering from temporal lobe epilepsy (TLE), and to describe the connection between LIL and other clinical ictal signs. We retrospectively reviewed video records of 87 patients with TLE. We reviewed a total of 276 focal epileptic seizures with or without secondary generalization. We studied the incidence of LIL, its lateralizing value, and its relationship to other ictal clinical signs. Of the 87 patients, 49 had undergone a successful resective surgery at least 1 year prior to the study. LIL is a late sign in the course of partial seizure. It occurred in 25 of our 87 patients (28.7%), and in 47 of 276 seizures (17.1%). In all of the evaluated seizures, LIL occurred contralateral to the side of seizure onset (P < 0.001). LIL was always associated with ipsilateral upper limb automatisms, and in 63.1% of the occurrences, it was immediately followed by ictal dystonia. LIL is a more accurate term to describe what has previously been called 'ictal paresis' in the literature. Due to the inability to execute proper testing during a partial seizure, it is better to use the term LIL when making a visual analysis of a seizure. LIL is a more suitable term to describe the studied ictal sign. It is a relatively frequent sign in patients with TLE. LIL has an excellent lateralizing value for the contralateral hemisphere. It is a negative motor sign, and its genesis is probably associated with the epileptic involvement of the contralateral frontal lobe. PMID- 16241979 TI - Lack of prefrontal repetitive transcranial magnetic stimulation effects in time production processing. AB - The aim of the present study was to determine the effects of high frequency repetitive transcranial magnetic stimulation (rTMS) over different neuroanatomical areas [left and right doroslateral prefrontal cortex (DLPFC) and right cerebellar hemisphere] on time production task. The study was performed in 16 healthy right-handed men with a cross-over, within subject repeated measures design. There were four rTMS conditions: baseline without stimulation, high frequency rTMS over right, left DLPFC and over right cerebellum. The volunteers were asked to produce a 3-min interval by internal counting. The rTMS was applied during the task. No significantly differences were observed in absolute error scores in time estimation task with any rTMS condition. This preliminary study does not support the role of the prefrontal lobe in time production processes. PMID- 16241980 TI - Epidemiology of status epilepticus in a rural area of northern Italy: a 2-year population-based study. AB - We performed a 2-year population-based study on status epilepticus (SE) in adults in the rural area of Lugo di Romagna, northern Italy, to verify whether an area of low-level urbanization has a lower risk of occurrence of SE (as recently suggested), different clinical features and short-term prognosis than areas of high-level urbanization. We found crude and age- and sex-adjusted annual incidence rates of SE of 16.5/100 000 and 11.6/100 000, respectively. In patients under 60 years crude incidence was 2.9/100 000 and in the elderly (>/=60 years) 38.6/100 000. Acute symptomatic SE accounted for 30% and a cerebrovascular pathology was the most frequently associated etiologic condition (60%). A history of seizures was reported in 41% of patients. The first therapeutic intervention was mainly benzodiazepines (lorazepam 46%; diazepam 33%). The 30-day case fatality was 7%. We observed that the adult population of an area with a low level of urbanization has the same risk for SE, clinical features and short-term prognosis as European urban areas. The only contrasting result is the 30-day case fatality of 7% against the 39% found in the other Italian study (Bologna), despite the similarity of the SE features in these two areas of the same region. We infer that the short-term prognosis of SE could also be considerably influenced by differences in health service organization (and hence management) possibly due to different levels of urbanization. PMID- 16241981 TI - Seasonal, extratrigeminal, episodic paroxysmal hemicrania successfully treated with single suboccipital steroid injections. AB - This case report describes a case of extratrigeminal, episodic paroxysmal hemicrania with a clear seasonal temporal pattern, successfully treated with repeated single suboccipital steroid injections. The pathophysiological and clinical implications of this observation are discussed. PMID- 16241982 TI - Spelling tasks and Alzheimer's disease staging. AB - Reading, writing and oral spelling were evaluated in 30 normal elderly individuals and 28 patients with probable Alzheimer's disease (AD). Significant differences between control and AD groups were found in all tasks. Oral spelling was the most impaired function in AD and the only one which differentiated mild and moderate dementia cases. This task is very sensitive to the disease's effects on working memory and may be included in batteries aimed at staging AD. PMID- 16241983 TI - X-linked adrenoleukodystrophy with olivopontocerebellar atrophy. AB - X-linked adrenoleukodystrophy (X-ALD) is a rare neurological disorder characterized by adrenal, gonadal and nervous system dysfunction. Patients usually develop spinal cord degeneration with involvement of the cerebral white matter. While a spinocerebellar variant has been described, the selective involvement of cerebellar white matter is very rare. We report the case of a patient affected by X-ALD whose clinical and magnetic resonance imaging (MRI) results resembled olivopontocerebellar atrophy. He was a 29-year-old mentally retarded man, who began to complain of slowly progressive gait ataxia after an 8 year history of Addison's disease. Serial MRI revealed marked cerebellar atrophy involving the inferior cerebellar vermis and brainstem, but sparing the supratentorial white matter. The diagnosis of X-ALD was confirmed by elevated levels of very long-chain fatty acids in the serum. After 2 years follow-up, the patient developed spastic paraparesis. The patient represents an unusual clinical presentation of X-ALD, as further confirmed by the MRI results. Consequently, cerebellar symptoms should be considered as a clinical presentation of X-ALD. Early recognition of this rare disorder would be useful for genetic counselling and therapy. PMID- 16241984 TI - Comparison of immunomodulatory treatments for multiple sclerosis. PMID- 16241986 TI - Hass J, Firzlaff M (2005). Twenty-four-month comparison of immunomodulatory treatments--a retrospective open label study in 308 RRMS patients treated with beta interferons or glatiramer acetate (Copaxone). European Journal of Neurology 12:425-431. PMID- 16241992 TI - Breast pathology practice: most common problems in a consultation service. AB - Considerable progress has been made in understanding breast lesions utilizing molecular methods, but conventional morphology, simple immunohistochemical stains and common sense still prevail in diagnosing the vast majority of breast disease. The focus of this review is to identify the most common breast lesions sent to our consultation practice, and to reiterate salient diagnostic features, differential diagnoses and common pitfalls in identifying these lesions. Separation of epithelial proliferative lesions and differentiation between usual epithelial hyperplasia (UEH) and atypical ductal hyperplasia (ADH) are the most common problems encountered in our Consultation practice. Differentiation between UEH and ADH is based on the assumption that ADH is a clonal process, recognized by a uniform phenotype and more recently described immunohistochemical markers such as differential cytokeratin and also hormone receptor expression. Difficulty in subtyping invasive carcinomas and exclusion of in situ and/or invasive carcinoma in a sclerosing lesion is also commonly noted. Finally, problems in distinguishing various papillary and fibroepithelial lesions are also encountered. The use of common immunohistochemical stains such as various cytokeratin and myoepithelial markers, E-cadherin and hormone receptors is helpful in solving most of these diagnostic dilemmas. PMID- 16241993 TI - p63 correlates with both BRCA1 and cytokeratin 5 in invasive breast carcinomas: further evidence for the pathogenesis of the basal phenotype of breast cancer. AB - AIMS: To study the expression of p63, cytokeratin (CK) 5 and CK8/18 in invasive ductal carcinomas and their relationship with BRCA1 and other pathological and immunohistochemical features of clinical significance. METHODS AND RESULTS: Immunohistochemistry with the antibodies p63, CK5, CK8/18, BRCA1, oestrogen receptor, progesterone receptor, p53, c-erbB-2 and Ki67 was performed in 102 formalin-fixed paraffin-embedded samples of invasive ductal carcinomas. The CK5+ cases were submitted to a double-immunolabelling study with p63. There was a strong relationship between CK5 and p63 expression and both markers were associated with hormonal receptor-negative high-grade carcinomas with high proliferative rate. Furthermore, there was coexpression of CK5 and p63 in neoplastic cells, indicating that p63, like CK5, is a marker of the basal phenotype of breast cancer. There was a strong relationship between reduced expression of BRCA1 with both p63 and CK5 expression as well as an inverse correlation between p63 and CK8/18 expression, suggesting that loss of p63 expression is required for the transition between a basal to a luminal phenotype of breast carcinoma. CONCLUSIONS: Since p63 is thought to be a marker of stem cells and may act as an oncogene, our data support the idea that BRCA1 acts as stem cell regulator. PMID- 16241994 TI - Lymphomatous polyposis of the gastrointestinal tract, including mantle cell lymphoma, follicular lymphoma and mucosa-associated lymphoid tissue lymphoma. AB - AIMS: Lymphomatous polyposis (LP) is considered to represent mantle cell lymphoma (MCL) of the gastrointestinal (GI) tract. However, a few reports have suggested that some are follicular lymphoma (FL) or mucosa-associated lymphoid tissue (MALT) lymphomas. In this study, we analysed 35 patients and clarified the clinicopathological features of LP. METHODS AND RESULTS: Paraffin-embedded tissue samples were stained immunohistochemically and analysed by tissue-fluorescence in situ hybridization (T-FISH) for IGH/CCND1 (cyclin D1) and IGH/BCL2. The average age of the patients was 58.3 years. Over half of the cases showed gastric, duodenal, small intestinal, ileocaecal and sigmoid colonic lesions (15, 19, 15, 16 and 16 cases, respectively). Phenotypically, cases were classified into three types of MCL (cyclin D1+ CD5+ CD10-) (n=12), FL (cyclin D1- CD5- CD10+) (n=14) and MALT (cyclin D1- CD5- CD10-) (n=9). T-FISH identified 11 of the 11 examined cases with MCLs to have IGH/CCND1, while seven of 10 cases with FL had IGH/BCL2, and none of the MALT cases were positive for IGH/CCND1 or IGH/BCL2. At the study endpoint, five of 12 patients with MCL were dead, two of 14 with FL and one of nine with MALT were dead of other disease. Event-free survival analysis showed significantly poorest outcome in MCL, followed by FL, while MALT was associated with a favourable outcome (P=0.0040). CONCLUSIONS: Our study emphasizes the importance of differentiating MCL, FL and MALT of LP in evaluating prognosis and hence the most suitable therapeutic regimen. PMID- 16241995 TI - Primary invasive micropapillary carcinoma of the colon. AB - AIMS: Invasive micropapillary carcinoma (IMPC) is associated with frequent lymph node metastasis and adverse clinical outcome. IMPC has been reported in breast, urinary bladder, ureter, lung and parotid gland but not in colon. We present the clinicopathological features of three cases of primary IMPC of the colon with a review of the literature. METHODS AND RESULTS: The patients (one man and two women) were 53, 67 and 68 years old, respectively. The size of the tumour ranged from 20 to 100 mm in diameter. Histologically, all cases were composed predominantly of papillary tumour cell clusters with spaces in a background of fine fibrocollagenous stroma. One of the tumours (case 1) was nearly completely composed of IMPC, but the other two were associated with foci of adenocarcinoma and concurrent mucinous carcinoma, respectively. MUC1 was positive in all cases, suggestive of reverse cell orientation which is responsible for its unique histological features. CONCLUSIONS: We report three cases of primary IMPC of the colon. Its clinical significance remains undetermined but the presence of this component may represent a poor prognostic factor. PMID- 16241996 TI - Histopathology of acute acalculous cholecystitis in critically ill patients. AB - AIMS: To illustrate the histopathological features of acute acalculous cholecystitis (AAC) of critically ill patients and to compare them with those of acute calculous cholecystitis (ACC) and normal gallbladders. METHODS AND RESULTS: We studied 34 gallbladders with AAC and compared them with 28 cases of ACC and 14 normal gallbladders. Histological features were systematically evaluated. Typical features in AAC were bile infiltration, leucocyte margination of blood vessels and lymphatic dilation. Bile infiltration in the gallbladder wall was more common and extended wider and deeper into the muscle layer in AAC compared with ACC. Epithelial degeneration and defects and widespread occurrence of inflammatory cells were typical features in ACC. Necrosis in the muscle layer was also more common and extended wider and deeper in ACC. There were no differences in the occurrence of capillary thromboses, lymphatic follicles or Rokitansky-Aschoff sinuses between the AAC and ACC samples. CONCLUSIONS: There are characteristic differences in histopathology between AAC and ACC, although due to overlap, none appeared to be specific as such for either condition. These results suggest that AAC is largely a manifestation of systemic critical illness, whereas ACC is a local disease of the gallbladder. PMID- 16241997 TI - Lymphocyte subsets in irradiation-induced sialadenitis of the submandibular gland. AB - AIMS: Irradiation-induced sialadenitis is a significant cause of morbidity in head and neck cancer patients receiving radiotherapy. Neither the exact aetiopathology of chronic irradiation-induced sialadenitis nor the mechanisms leading to atrophy of the glandular cells associated with an increase in extracellular matrix are understood. The aim of our study was to determine the phenotype of the inflammatory infiltrate and to study its distribution in the affected submandibular glands. METHODS AND RESULTS: Paraffin-embedded submandibular glands from a homogeneous group of 19 patients with advanced oropharyngeal cancer who received conventional radiotherapy to the primary site and upper neck were analysed. In all patients the radiation dose and field were approximately equal. The submandibular glands were obtained during neck dissection. To characterize the lymphoid infiltrate, all tissue sections were immunostained for T cells (CD3, CD4, CD8), cytotoxic T cells (granzyme B), B cells (CD20), and macrophages (Ki-M1p). A histopathological classification into four grades was established based on the degree of glandular atrophy, fibrosis and lymphocytic infiltration. Phenotypic analysis of submandibular gland sections revealed that the great majority of lymphocytic infiltrates were cytotoxic T cells associated with acinar cell destruction. CONCLUSIONS: The significantly elevated frequencies of cytotoxic cells in the submandibular glands of patients with irradiation-induced sialadenitis suggest that cell-mediated immune mechanisms may play a part in the pathogenesis of this disease. PMID- 16241998 TI - Expression of acyl-CoA synthetase 5 in human endometrium and in endometrioid adenocarcinomas. AB - AIM: Fatty acid metabolism of the endometrium is important for tissue homeostasis in the proliferative and secretory phase of the menstrual cycle. The enzyme acyl CoA synthetase 5 (ACS5) plays a crucial role in fatty acid metabolism, mainly through the generation of multifunctional long-chain-fatty-acid-CoA esters. The aim of the present study was to characterize expression and localization of ACS5 in the normal human endometrium and in endometrioid adenocarcinomas. METHODS AND RESULTS: Expression of ACS5 in the human endometrium was investigated by in situ techniques (immunohistochemistry, mRNA in situ hybridization) and a molecular approach (reverse transcriptase-polymerase chain reaction, Western blot). ACS5 protein and mRNA were localized to the epithelium of the human endometrium. Here, ACS5 expression was found throughout the menstrual cycle as well as in the postmenopausal endometrium. Notably, in endometrioid adenocarcinomas, the ACS5 molecule was found abundantly in well-differentiated tumours, but not in poorly differentiated adenocarcinomas. CONCLUSIONS: The abundance of ACS5 in the endometrial epithelium throughout the menstrual cycle provides support for its role in the regulation of tissue homeostasis. With regard to its value for histopathological diagnosis, immunohistochemical characterization of endometrioid adenocarcinomas shows that a decrease in ACS5 expression correlates with tumour dedifferentiation. PMID- 16241999 TI - Histopathological features of chronic granulomatous disease (CGD) in childhood. AB - AIMS: To describe the spectrum of histopathological features encountered in children with chronic granulomatous disease (CGD) at a specialist centre. METHODS AND RESULTS: The histopathological findings of 88 surgical pathology requests from a range of organ systems including upper and lower gastrointestinal tract biopsy series, liver, bladder, bone, lung, skin, soft tissue, bone marrow and lymph node biopsy specimens, in 32 patients aged 4 months to 18 years (median 7 years) with CGD were reviewed. In most tissues the features were those of active chronic inflammation, with or without abscess or granuloma formation, often associated with fungal infection. In some tissues, more characteristic findings were identified, including the presence of pigmented macrophages, especially in hepatic sinusoids and colonic mucosa, where active chronic eosinophil predominant colitis was also observed. CONCLUSIONS: Chronic granulomatous disease may present to histopathologists in a wide range of tissue specimens most often demonstrating features of active chronic inflammation with or without granuloma formation. The presence of numerous pigmented macrophages in association with such inflammation should raise suspicion of the diagnosis. In addition, diffuse granulomatous inflammation of the lung and hepatic abscess formation should be regarded as suggestive of the diagnosis. PMID- 16242000 TI - CD117+ small cell lung cancer lacks the asp 816-->val point mutation in exon 17. AB - AIMS: To determine the frequency of point mutation in c-kit in CD117+ small cell lung cancer (SCLC). A significant proportion of SCLCs have been documented to be CD117+, thereby signifying they express the c-kit gene product. This finding suggests this tumour may be a potential target for tyrosine kinase inhibitor (TKI) agents directed at c-kit. A point mutation in exon 17 of the c-kit gene, however, can abrogate the binding of TKIs. This being the case, immunohistochemistry is necessary to identify potential candidates for treatment with TKIs, but DNA sequence analysis may need to be performed to determine if these tumours will respond. METHODS AND RESULTS: Tumour cells of 23 cases of SCLC showing immunoreactivity for CD117 were laser capture microdissected from archived formalin-fixed paraffin-embedded tissue and the DNA isolated. PCR on exon 17 of the c-kit gene was performed and the amplified product sequenced. No point mutations were detected. CONCLUSIONS: The absence of mutations in exon 17 of CD117+ SCLC suggests this tumour may respond to therapy with TKI. PMID- 16242001 TI - Primary lymphoma arising in the nasal cavity among Japanese. AB - AIMS: Most lymphomas arising in the nasal cavity are thought to be of natural killer (NK) cell origin. However, some reports indicate that T- and B-cell lymphomas may also primarily arise in the nasal cavity. We therefore studied lymphomas arising in the nasal cavity both histologically and immunohistochemically. METHODS AND RESULTS: Of the 32 cases investigated, 20 cases were also available as fresh frozen specimens. We diagnosed 31 cases as extranodal NK/T-cell lymphoma and one as plasmacytoma. The neoplastic cells were immunoreactive for CD3 (polyclonal) 31/31, LMP-1 12/31, CD20 (L26) 0/31, granzyme B 30/31, TIA-1 30/30, CD56 (123C3) 29/31, CD4 0/31 and CD8 3/31. In situ hybridization for Epstein-Barr virus-encoded small RNA-1 (EBER-1) was detected in 31/31. In frozen tissue sections, neoplastic cells mostly showed CD3 (Leu4)-, CD4 (Leu3a)-, CD5 (Leu1)-, CD8 (Leu2)-, CD16 (Leu11)-, CD56 (Leu19)+, betaF1-, TCRdelta1-, perforin+, CD94+ phenotypes. These immunohistochemical findings indicate their NK cell origin. In three cases, neoplastic cells were positive for CD8. In one of these cases, neoplastic cells were positive for CD8beta and Valpha24, suggesting their NKT-cell origin. CONCLUSIONS: Our present study indicates that primary lymphomas arising in the nasal cavity are mostly of NK cell derivation. Our present study also suggests that a small number of cases are derived from NKT-cells. PMID- 16242003 TI - Toluidine positivity in chromophobe renal cell carcinoma: use in intraoperative diagnosis. PMID- 16242004 TI - Chordomas do not express CD10 and renal cell carcinoma (RCC) antigen: an immunohistochemical study. PMID- 16242005 TI - Ampullary carcinoid with amyloid stroma. PMID- 16242006 TI - Squamous inclusion cyst with evidence of focal glandular differentiation in an axillary lymph node. PMID- 16242007 TI - Uterine leiomyosarcoma with HMB45+ clear cell areas: report of two cases. PMID- 16242008 TI - An exotic osteoclastic reaction to breast implant that mimicked breast carcinoma. PMID- 16242009 TI - Measurement of whole tumour size in breast cancer specimens. PMID- 16242010 TI - Asymptomatic dissecting aneurysm of the coeliac artery: a variant of segmental arterial mediolysis. PMID- 16242011 TI - Simultaneous non-Hodgkin's lymphoma and mesothelioma presenting as a collision tumour. PMID- 16242013 TI - The burden of diarrhoea, shigellosis, and cholera in North Jakarta, Indonesia: findings from 24 months surveillance. AB - BACKGROUND: In preparation of vaccines trials to estimate protection against shigellosis and cholera we conducted a two-year community-based surveillance study in an impoverished area of North Jakarta which provided updated information on the disease burden in the area. METHODS: We conducted a two-year community based surveillance study from August 2001 to July 2003 in an impoverished area of North Jakarta to assess the burden of diarrhoea, shigellosis, and cholera. At participating health care providers, a case report form was completed and stool sample collected from cases presenting with diarrhoea. RESULTS: Infants had the highest incidences of diarrhoea (759/1,000/year) and cholera (4/1,000/year). Diarrhea incidence was significantly higher in boys under 5 years (387/1,000/year) than girls under 5 years (309/1,000/year; p < 0.001). Children aged 1 to 2 years had the highest incidence of shigellosis (32/1,000/year). Shigella flexneri was the most common Shigella species isolated and 73% to 95% of these isolates were resistant to ampicillin, trimethoprim-sulfamethoxazole, chloramphenicol and tetracycline but remain susceptible to nalidixic acid, ciprofloxacin, and ceftriaxone. We found an overall incidence of cholera of 0.5/1,000/year. Cholera was most common in children, with the highest incidence at 4/1,000/year in those less than 1 year of age. Of the 154 V. cholerae O1 isolates, 89 (58%) were of the El Tor Ogawa serotype and 65 (42%) were El Tor Inaba. Thirty-four percent of patients with cholera were intravenously rehydrated and 22% required hospitalization. V. parahaemolyticus infections were detected sporadically but increased from July 2002 onwards. CONCLUSION: Diarrhoea causes a heavy public health burden in Jakarta particularly in young children. The impact of shigellosis is exacerbated by the threat of antimicrobial resistance, whereas that of cholera is aggravated by its severe manifestations. PMID- 16242014 TI - Status of national research bioethics committees in the WHO African region. AB - BACKGROUND: The Regional Committee for Africa of the World Health Organization (WHO) in 2001 expressed concern that some health-related studies undertaken in the Region were not subjected to any form of ethics review. In 2003, the study reported in this paper was conducted to determine which Member country did not have a national research ethics committee (REC) with a view to guiding the WHO Regional Office in developing practical strategies for supporting those countries. METHODS: This is a descriptive study. The questionnaire was prepared and sent by diplomatic pouch to all the 46 Member States in the WHO African Region, through the WHO country representatives, for facilitation and follow up. The data were entered in Excel spreadsheet and subsequently exported to STATA for analysis. A Chi-Squared test (chi2) for independence was undertaken to test the relationship between presence/absence of Research Ethics Committee (REC) and selected individual socioeconomic and health variables. RESULTS: The main findings were as follows: the response rate was 61% (28/46); 64% (18/28) confirmed the existence of RECs; 36% (10/28) of the respondent countries did not have a REC (although 80% of them reported that they had in place an ad hoc ethical review mechanism); 85% (22/26) of the countries that responded to this question indicated that ethical approval of research proposals was, in principle, required; and although 59% of the countries that had a REC expected it to meet every month, only 44% of them reported that the REC actually met on a monthly basis. In the Chi-Squared test, only the average population in the group of countries with a REC was statistically different (at 5% level of significance) from that of the group of countries without a REC. CONCLUSION: In the current era of globalized biomedical research, good ethics stewardship demands that every country, irrespective of its level of economic development, should have in place a functional research ethics review system in order to protect the dignity, integrity and safety of its citizens who participate in research. PMID- 16242015 TI - Endospores of halophilic bacteria of the family Bacillaceae isolated from non saline Japanese soil may be transported by Kosa event (Asian dust storm). AB - BACKGROUND: Generally, extremophiles have been deemed to survive in the extreme environments to which they had adapted to grow. Recently many extremophiles have been isolated from places where they are not expected to grow. Alkaliphilic microorganisms have been isolated from acidic soil samples with pH 4.0, and thermophiles have been isolated from samples of low temperature. Numerous moderately halophilic microorganisms, defined as those that grow optimally in media containing 0.5-2.5 Molar (3-15%) NaCl, and halotolerant microorganisms that are able to grow in media without added NaCl and in the presence of high NaCl have been isolated from saline environments such as salterns, salt lakes and sea sands. It has tacitly been believed that habitats of halophiles able to grow in media containing more than 20% (3.4 M) are restricted to saline environments, and no reports have been published on the isolation of halophiles from ordinary garden soil samples. RESULTS: We demonstrated that many halophilic bacteria that are able to grow in the presence of 20% NaCl are inhabiting in non-saline environments such as ordinary garden soils, yards, fields and roadways in an area surrounding Tokyo, Japan. Analyses of partial 16S rRNA gene sequences of 176 isolates suggested that they were halophiles belonging to genera of the family Bacillaceae, Bacillus (11 isolates), Filobacillus (19 isolates), Gracilibacillus (6 isolates), Halobacillus (102 isolates), Lentibacillus (1 isolate), Paraliobacillus (5 isolates) and Virgibacillus (17 isolates). Sequences of 15 isolates showed similarities less than 92%, suggesting that they may represent novel taxa within the family Bacillaceae. CONCLUSION: The numbers of total bacteria of inland soil samples were in a range from 1.4 x 10(7)/g to 1.1 x 10(6)/g. One tenth of the total bacteria was occupied by endospore-forming bacteria. Only very few of the endospore-forming bacteria, roughly 1 out of 20,000, are halophilic bacteria. Most of the halophilic bacteria were surviving as endospores in the soil samples, in a range of less than 1 to about 500/g soil. Samples collected from seashore in a city confronting Tokyo Bay gave the total numbers of bacteria and endospores roughly 1000 time smaller than those of inland soil samples. Numbers of halophilic bacteria per gram, however, were almost the same as those of inland soil samples. A possible source of the halophilic endospore originating from Asian dust storms is discussed. PMID- 16242016 TI - Cardiac involvement in a patient with clinical and serological evidence of African tick-bite fever. AB - BACKGROUND: Myocarditis and pericarditis are rare complications of rickettsiosis, usually associated with Rickettsia rickettsii and R. conorii. African tick-bite fever (ATBF) is generally considered as a benign disease and no cases of myocardial involvement due to Rickettsia africae, the agent of ATBF, have yet been described. CASE PRESENTATION: The patient, that travelled in an endemic area, presented typical inoculation eschars, and a seroconversion against R. africae, was admitted for chest pains and increased cardiac enzymes in the context of an acute myocarditis. CONCLUSION: Our findings suggest that ATBF, that usually presents a benign course, may be complicated by an acute myocarditis. PMID- 16242017 TI - Research influence on antimalarial drug policy change in Tanzania: case study of replacing chloroquine with sulfadoxine-pyrimethamine as the first-line drug. AB - INTRODUCTION: Research is an essential tool in facing the challenges of scaling up interventions and improving access to services. As in many other countries, the translation of research evidence into drug policy action in Tanzania is often constrained by poor communication between researchers and policy decision-makers, individual perceptions or attitudes towards the drug and hesitation by some policy decision-makers to approve change when they anticipate possible undesirable repercussions should the policy change as proposed. Internationally, literature on the role of researchers on national antimalarial drug policy change is limited. OBJECTIVES: To describe the (a) role of researchers in producing evidence that influenced the Tanzanian government replace chloroquine (CQ) with sulfadoxine-pyrimethamine (SP) as the first-line drug and the challenges faced in convincing policy-makers, general practitioners, pharmaceutical industry and the general public on the need for change (b) challenges ahead before a new drug combination treatment policy is introduced in Tanzania. METHODS: In-depth interviews were held with national-level policy-makers, malaria control programme managers, pharmaceutical officers, general medical practitioners, medical research library and publications officers, university academicians, heads of medical research institutions and district and regional medical officers. Additional data were obtained through a review of malaria drug policy documents and participant observations were also done. RESULTS: In year 2001, the Tanzanian Government officially changed its malaria treatment policy guidelines whereby CQ- the first-line drug for a long time was replaced with SP. This policy decision was supported by research evidence indicating parasite resistance to CQ and clinical CQ treatment failure rates to have reached intolerable levels as compared to SP and amodiaquine (AQ). Research also indicated that since SP was also facing rising resistance trend, the need for a more effective drug was indispensable but for an interim 5-10 year period it was justifiable to recommend SP that was relatively more cost-effective than CQ and AQ. The government launched the policy change considering that studies (ethically approved by the Ministry of Health) on therapeutic efficacy and cost-effectiveness of artemisinin drug combination therapies were underway. Nevertheless, the process of communicating research results and recommendations to policy-making authorities involved critical debates between policy makers and researchers, among the researchers themselves and between the researchers and general practitioners, the speculative media reports on SP side-effects and reservations by the general public concerning the rationale for policy change, when to change, and to which drug of choice. CONCLUSION: Changing national drug policy will remain a sensitive issue that cannot be done overnight. However, to ensure that research findings are recognised and the recommendations emanating from such findings are effectively utilized, a systematic involvement of all the key stakeholders (including policy-makers, drug manufacturers, media, practitioners and the general public) at all stages of research is crucial. It also matters how and when research information is communicated to the stakeholders. Professional organizations such as the East African Network on Malaria Treatment have potential to bring together malaria researchers, policy-makers and other stakeholders in the research-to-drug policy change interface. PMID- 16242018 TI - HDL enhances oxidation of LDL in vitro in both men and women. AB - BACKGROUND: Oxidative modification of low-density lipoprotein (LDL) is a key event in the oxidation hypothesis of atherogenesis. Some in vitro experiments have previously suggested that high-density lipoprotein (HDL) co-incubated with LDL prevents Cu2+-induced oxidation of LDL, while some other studies have observed an opposite effect. To comprehensively clarify the role of HDL in this context, we isolated LDL, HDL2 and HDL3 from sera of 61 free-living individuals (33 women and 28 men). RESULTS: When the isolated LDL was subjected to Cu2+ induced oxidation, both HDL2 and HDL3 particles increased the rate of appearance and the final concentration of conjugated dienes similarly in both genders. Oxidation rate was positively associated with polyunsaturated fatty acid content of the lipoproteins in that it was positively related to the content of linoleate and negatively related to oleate. More saturated fats thus protected the lipoproteins from damage. CONCLUSION: We conclude that in vitro HDL does not protect LDL from oxidation, but is in fact oxidized fastest of all lipoproteins due to its fatty acid composition, which is oxidation promoting. PMID- 16242019 TI - nfi-I affects behavior and life-span in C. elegans but is not essential for DNA replication or survival. AB - BACKGROUND: The Nuclear Factor I (one) (NFI) family of transcription/replication factors plays essential roles in mammalian gene expression and development and in adenovirus DNA replication. Because of its role in viral DNA replication NFI has long been suspected to function in host DNA synthesis. Determining the requirement for NFI proteins in mammalian DNA replication is complicated by the presence of 4 NFI genes in mice and humans. Loss of individual NFI genes in mice cause defects in brain, lung and tooth development, but the presence of 4 homologous NFI genes raises the issue of redundant roles for NFI genes in DNA replication. No NFI genes are present in bacteria, fungi or plants. However single NFI genes are present in several simple animals including Drosophila and C. elegans, making it possible to test for a requirement for NFI in multicellular eukaryotic DNA replication and development. Here we assess the functions of the single nfi-1 gene in C. elegans. RESULTS: C. elegans NFI protein (CeNFI) binds specifically to the same NFI-binding site recognized by vertebrate NFIs. nfi-1 encodes alternatively-spliced, maternally-inherited transcripts that are expressed at the single cell stage, during embryogenesis, and in adult muscles, neurons and gut cells. Worms lacking nfi-1 survive but have defects in movement, pharyngeal pumping and egg-laying and have a reduced life-span. Expression of the muscle gene Ce titin is decreased in nfi-1 mutant worms. CONCLUSION: NFI gene function is not needed for survival in C. elegans and thus NFI is likely not essential for DNA replication in multi-cellular eukaryotes. The multiple defects in motility, egg-laying, pharyngeal pumping, and reduced lifespan indicate that NFI is important for these processes. Reduction in Ce titin expression could affect muscle function in multiple tissues. The phenotype of nfi-1 null worms indicates that NFI functions in multiple developmental and behavioral systems in C. elegans, likely regulating genes that function in motility, egg-laying, pharyngeal pumping and lifespan maintenance. PMID- 16242021 TI - Long-term all-sites cancer mortality time trends in Ohio, USA, 1970-2001: differences by race, gender and age. AB - BACKGROUND: There were significant changes in cancer mortality in the USA over the last several decades, in the whole country and in particular states. However, no in depth analysis has been published so far, dealing with changes in mortality time trends in the state of Ohio. Since the state of Ohio belongs to the states of relatively high level of all-sites mortality in both males and females, it is of interest to analyze recent changes in mortality rates, as well as to compare them with the situation in the rest of the USA. The main aim of this study was to analyze, describe and interpret all-sites cancer mortality time trends in the population of the State of Ohio. METHODS: Cancer mortality data by age, sex, race and year for the period 1970-2001 were obtained from the Surveillance Research Program of the National Cancer Institute SEER*Stat software. A joinpoint regression methodology was used to provide estimated annual percentage changes (EAPCs) and to detect points in time where significant changes in the trends occurred. RESULTS: In both, males and females mortality rates were higher in blacks compared with whites. The difference was bigger in males (39.9%) than in women (23.3%). Mortality rates in Ohio are generally higher than average USA rates--an overall difference was 7.5% in men in 1997-2001, and 6.1% in women. All sites mortality trends in Ohio and in the whole USA are similar. However, in general, mortality rates in Ohio remained elevated compared with the USA rates throughout the entire analyzed period. The exceptions are the rates in young and middle-aged African Americans. CONCLUSION: Although direction of time trends in Ohio are similar in Ohio and the whole US, Ohio still have cancer mortality rates higher than the US average. In addition, there is a significant discrepancy between white and black population of Ohio in all-sites mortality level, with disadvantage for Blacks. To diminish disparities in cancer mortality between African Americans and white inhabitants of Ohio efforts should be focused on increasing knowledge of black people regarding healthy lifestyle and behavioral risk factors, but also on diminishing socioeconomic differences, and last but not least, on better access to medical care. PMID- 16242020 TI - Comparative genomics of Thermus thermophilus and Deinococcus radiodurans: divergent routes of adaptation to thermophily and radiation resistance. AB - BACKGROUND: Thermus thermophilus and Deinococcus radiodurans belong to a distinct bacterial clade but have remarkably different phenotypes. T. thermophilus is a thermophile, which is relatively sensitive to ionizing radiation and desiccation, whereas D. radiodurans is a mesophile, which is highly radiation- and desiccation resistant. Here we present an in-depth comparison of the genomes of these two related but differently adapted bacteria. RESULTS: By reconstructing the evolution of Thermus and Deinococcus after the divergence from their common ancestor, we demonstrate a high level of post-divergence gene flux in both lineages. Various aspects of the adaptation to high temperature in Thermus can be attributed to horizontal gene transfer from archaea and thermophilic bacteria; many of the horizontally transferred genes are located on the single megaplasmid of Thermus. In addition, the Thermus lineage has lost a set of genes that are still present in Deinococcus and many other mesophilic bacteria but are not common among thermophiles. By contrast, Deinococcus seems to have acquired numerous genes related to stress response systems from various bacteria. A comparison of the distribution of orthologous genes among the four partitions of the Deinococcus genome and the two partitions of the Thermus genome reveals homology between the Thermus megaplasmid (pTT27) and Deinococcus megaplasmid (DR177). CONCLUSION: After the radiation from their common ancestor, the Thermus and Deinococcus lineages have taken divergent paths toward their distinct lifestyles. In addition to extensive gene loss, Thermus seems to have acquired numerous genes from thermophiles, which likely was the decisive contribution to its thermophilic adaptation. By contrast, Deinococcus lost few genes but seems to have acquired many bacterial genes that apparently enhanced its ability to survive different kinds of environmental stresses. Notwithstanding the accumulation of horizontally transferred genes, we also show that the single megaplasmid of Thermus and the DR177 megaplasmid of Deinococcus are homologous and probably were inherited from the common ancestor of these bacteria. PMID- 16242023 TI - i-Tracker: for quantitative proteomics using iTRAQ. AB - BACKGROUND: iTRAQ technology for protein quantitation using mass spectrometry is a recent, powerful means of determining relative protein levels in up to four samples simultaneously. Although protein identification of samples generated using iTRAQ may be carried out using any current identification software, the quantitation calculations have been restricted to the ProQuant software supplied by Applied Biosciences. i-Tracker software has been developed to extract reporter ion peak ratios from non-centroided tandem MS peak lists in a format easily linked to the results of protein identification tools such as Mascot and Sequest. Such functionality is currently not provided by ProQuant, which is restricted to matching quantitative information to the peptide identifications from Applied Biosciences' Interrogator software. RESULTS: i-Tracker is shown to generate results that are consistent with those produced by ProQuant, thus validating both systems. CONCLUSION: i-Tracker allows quantitative information gained using the iTRAQ protocol to be linked with peptide identifications from popular tandem MS identification tools and hence is both a timely and useful tool for the proteomics community. PMID- 16242022 TI - The importance of immune gene variability (MHC) in evolutionary ecology and conservation. AB - Genetic studies have typically inferred the effects of human impact by documenting patterns of genetic differentiation and levels of genetic diversity among potentially isolated populations using selective neutral markers such as mitochondrial control region sequences, microsatellites or single nucleotide polymorphism (SNPs). However, evolutionary relevant and adaptive processes within and between populations can only be reflected by coding genes. In vertebrates, growing evidence suggests that genetic diversity is particularly important at the level of the major histocompatibility complex (MHC). MHC variants influence many important biological traits, including immune recognition, susceptibility to infectious and autoimmune diseases, individual odours, mating preferences, kin recognition, cooperation and pregnancy outcome. These diverse functions and characteristics place genes of the MHC among the best candidates for studies of mechanisms and significance of molecular adaptation in vertebrates. MHC variability is believed to be maintained by pathogen-driven selection, mediated either through heterozygote advantage or frequency-dependent selection. Up to now, most of our knowledge has derived from studies in humans or from model organisms under experimental, laboratory conditions. Empirical support for selective mechanisms in free-ranging animal populations in their natural environment is rare. In this review, I first introduce general information about the structure and function of MHC genes, as well as current hypotheses and concepts concerning the role of selection in the maintenance of MHC polymorphism. The evolutionary forces acting on the genetic diversity in coding and non-coding markers are compared. Then, I summarise empirical support for the functional importance of MHC variability in parasite resistance with emphasis on the evidence derived from free-ranging animal populations investigated in their natural habitat. Finally, I discuss the importance of adaptive genetic variability with respect to human impact and conservation, and implications for future studies. PMID- 16242024 TI - Missing effects of zinc in a porcine model of recurrent endotoxemia. AB - BACKGROUND: Chronic human sepsis often is characterised by the compensatory anti inflammatory response syndrome (CARS). During CARS, anti-inflammatory cytokines depress the inflammatory response leading to secondary and opportunistic infections. Proved in vitro as well as in vivo, zinc's pro-inflammatory effect might overcome this depression. METHODS: We used the model of porcine LPS-induced endotoxemia established by Klosterhalfen et al. 10 pigs were divided into two groups (n = 5). Endotoxemia was induced by recurrent intravenous LPS-application (1.0 microg/kg E. coli WO 111:B4) at hours 0, 5, and 12. At hour 10, each group received an intravenous treatment (group I = saline, group II = 5.0 mg/kg elementary zinc). Monitoring included hemodynamics, blood gas analysis, and the thermal dilution technique for the measurement of extravascular lung water and intrapulmonary shunt. Plasma concentrations of IL-6 and TNF-alpha were measured by ELISA. Morphology included weight of the lungs, width of the alveolar septae, and rate of paracentral liver necrosis. RESULTS: Zinc's application only trended to partly improve the pulmonary function. Compared to saline, significant differences were very rare. IL-6 and TNF-alpha were predominately measured higher in the zinc group. Again, significance was only reached sporadically. Hemodynamics and morphology revealed no significant differences at all. CONCLUSION: The application of zinc in this model of recurrent endotoxemia is feasible and without harmful effects. However, a protection or restoration of clinical relevance is not evident in our setting. The pulmonary function just trends to improve, cytokine liberation is only partly activated, hemodynamics and morphology were not influenced. Further pre-clinical studies have to define zinc's role as a therapeutic tool during CARS. PMID- 16242025 TI - Septal rupture with right ventricular wall dissection after myocardial infarction. AB - BACKGROUND: In patients with inferior myocardial infarction, septal rupture generally involves basal inferoposterior septum, and the communicating tract between left and right ventricle is often serpiginous with a variable degree of right ventricular wall extension. Right ventricular wall dissection following septal rupture related with previous myocardial infarction has been reported in a very few cases, in many of them this condition has been diagnosed in post-mortem studies. In a recent report long-term survival has been achieved after promptly echocardiographic diagnosis and surgical repair. CASE PRESENTATION: We present a case of a 59-year-old man who had a septal rupture with right ventricular wall dissection after inferior and right ventricular myocardial infarction. Transthoracic echocardiography, as first line examination, established the diagnosis, and prompt surgical repair allowed long-term survival in our patient. CONCLUSION: Outcomes after right ventricular intramyocardial dissection following septal rupture related to myocardial infarction has been reported to be dismal. Early recognition of this complication using transthoracic echocardiography at patient bedside, and prompt surgical repair are the main factors to achieve long term survival in these patients. PMID- 16242026 TI - The rectus sling to prevent loop colostomy retraction: a case series. AB - Diverting stomas are being used increasingly in the management of rectal cancer, particularly with low anterior resection following neoadjuvant therapy. We describe a simple anchorage method for loop colostomy using a rectus fascial sling. This has been used successfully in fifteen patients with no complications or evidence of significant spill over of faecal contents into the efferent loop. PMID- 16242027 TI - Systemic and local antibiotic prophylaxis in the prevention of Staphylococcus epidermidis graft infection. AB - BACKGROUND: The aim of the study was to investigate the in vivo efficacy of local and systemic antibiotic prophylaxis in the prevention of Staphylococcus (S.) epidermidis graft infection in a rat model and to evaluate the bacterial adherence to frequently used prosthetic graft materials. METHODS: Graft infections were established in the subcutaneous tissue of 120 male Wistar rats by implantation of Dacron/ePTFE grafts followed by topical inoculation with 2 x 10(7) CFUs of clinical isolate of methicillin-resistant S. epidermidis. Each of the graft series included a control group, one contaminated group that did not receive any antibiotic prophylaxis, two contaminated groups that received systemic prophylaxis with teicoplanin or levofloxacin and two contaminated groups that received teicoplanin-soaked or levofloxacin-soaked grafts. The grafts were removed 7 days after implantation and evaluated by quantitative culture. RESULTS: There was significant bacterial growth inhibition in the groups given systemic or local prophylaxis (P < 0.05). Methicillin-resistant S. epidermidis had greater affinity to Dacron graft when compared with ePTFE graft in the untreated contaminated groups (P < 0.05). CONCLUSION: The study demonstrated that the usage of systemic or local prophylaxis and preference of ePTFE graft can be useful in reducing the risk of vascular graft infections caused by staphylococcal strains with high levels of resistance. PMID- 16242028 TI - Massive lung collapse with partial resolution after several years: a case report. AB - BACKGROUND: Bronchitis obliterans is a severe and extremely rare complication of respiratory tract infections in children and is characterized by massive atelectasis and collapse of the affected lung. Of the rare cases reported in the literature all surviving children underwent surgical resection of the collapsed lung. CASE PRESENTATION: We report an infant with bronchitis obliterans that was treated conservatively. 5 years after the initial event, partial lung re expansion was documented. CONCLUSION: This case therefore supports a conservative treatment whenever possible with pneumonectomy only as a last treatment option. PMID- 16242029 TI - Ethnic variations in incidence of asthma episodes in England & Wales: national study of 502,482 patients in primary care. AB - BACKGROUND: Recent studies have demonstrated marked international variations in the prevalence of asthma, but less is known about ethnic variations in asthma epidemiology within individual countries and in particular the impact of migration on risk of developing asthma. Recent within country comparisons have however revealed that despite originating from areas of the world with a low risk for developing asthma, South Asian and Afro-Caribbean people in the UK are significantly (3x and 2x respectively) more likely to be admitted to hospital for asthma related problems than Whites. METHODS: Using data from the Fourth National Study of Morbidity Statistics in General Practice, a one-percent broadly representative prospective cohort study of consultations in general practice, we investigated ethnic variations in incident asthma consultations (defined as new or first consultations), and compared consultation rates between those born inside and outside the UK (migrant status). Logistic regression models were used to examine the combined effects of ethnicity and migration on asthma incident consultations. RESULTS: Results showed significantly lower new/first asthma consultation rates for Whites than for each of the ethnic minority groups studied (mean age-adjusted consultation rates per 1000 patient-years: Whites 26.4 (95%CI 26.4, 26.4); South Asians 30.4 (95%CI 30.3, 30.5); Afro-Caribbeans 35.1 (95%CI 34.9, 35.3); and Others 27.8 (27.7, 28.0). Within each of these ethnic groups, those born outside of the UK showed consistently lower rates of incident asthma consultations. Modelling the combined effects of ethnic and migrant status revealed that UK-born South Asians and Afro-Caribbeans experienced comparable risks for incident GP consultations for asthma to UK-born Whites. Non-UK born Whites however experienced reduced risks (adjusted OR 0.82, 95%CI 0.69, 0.97) whilst non-UK born South Asians experienced increased risks (adjusted OR 1.33, 95%CI 1.04, 1.70) compared to UK-born Whites. CONCLUSION: These findings strongly suggest that ethnicity and migration have significant and independent effects on asthma incidence. The known poorer asthma outcomes in UK South Asians and Afro Caribbeans may in part be explained by the offspring of migrants experiencing an increased risk of developing asthma when compared to UK-born Whites. This is the first study to find heterogeneity for incident asthma consultations in Whites by migrant status. PMID- 16242030 TI - Women's autonomy, education and contraception use in Pakistan: a national study. AB - BACKGROUND: It has been proposed that the autonomy of women is one of the mechanisms of how education influences contraceptive use in developing countries. We tested this hypothesis in a national sample of women in Pakistan. METHODS: We used the 2000 Pakistan Reproductive Health and Family Planning Survey, which interviewed a national sample of ever married women aged 15-49 years (n = 6579). Women's decision autonomy was estimated from 9 questions on who makes decisions at home; movement autonomy was based on 6 questions on whether women need permission to visit places outside home. A number of socio-demographic variables were used in multivariate analysis to investigate the independent association between autonomy and lifetime and current contraception use and to assess the extent to which autonomy mediates the association between education and contraception use. RESULTS: Decision autonomy was significantly associated with both lifetime and current contraception use; after controlling for covariates, the odds ratios for the highest vs. the lowest quintile were 1.8 (1.4-2.4) and 2.0 (1.4-2.8), respectively. Movement autonomy was not consistently associated with contraceptive use. Contraceptive use was strongly associated with women's education but this relation was not mediated by women's autonomy. CONCLUSION: Women's decision autonomy is significantly associated with contraceptive use but it does not appear to mediate the link between woman's education and contraception. PMID- 16242031 TI - Asymmetry in the burial of hydrophobic residues along the histone chains of eukarya, archaea and a transcription factor. AB - BACKGROUND: The histone fold is a common structural motif of proteins involved in the chromatin packaging of DNA and in transcription regulation. This single chain fold is stabilized by either homo- or hetero-dimer formation in archaea and eukarya. X-ray structures at atomic resolution have shown the eukaryotic nucleosome core particle to consist of a central tetramer of two bound H3-H4 dimers flanked by two H2A-H2B dimers. The c-terminal region of the H3 histone fold involved in coupling the two eukaryotic dimers of the tetramer, through a four-fold helical bundle, had previously been shown to be a region of reduced burial of hydrophobic residues within the dimers, and thereby provide a rationale for the observed reduced stability of the H3-H4 dimer compared with that of the H2A-H2B dimer. Furthermore, comparison between eukaryal and archaeal histones had suggested that this asymmetry in the distribution of hydrophobic residues along the H3 histone chains could be due to selective evolution that enhanced the coupling between the eukaryotic dimers of the tetramer. RESULTS AND DISCUSSION: The present work describes calculations utilizing the X-ray structures at atomic resolution of a hyperthermophile from Methanopyrus kandleri (HMk) and a eukaryotic transcription factor from Drosophila melanogaster (DRm), that are structurally homologous to the eukaryotic (H3-H4)2 tetramer. The results for several other related structures are also described. Reduced burial of hydrophobic residues, at the homologous H3 c-terminal regions of these structures, is found to parallel the burial at the c-terminal regions of the H3 histones and is, thereby, expected to affect dimer stability and the processes involving histone structural rearrangement. Significantly different sequence homology between the two histones of the HMk doublet with other archaeal sequences is observed, and how this might have occurred during selection to enhance tetramer stability is described. PMID- 16242032 TI - Early relief of osteoarthritis symptoms with a natural mineral supplement and a herbomineral combination: a randomized controlled trial [ISRCTN38432711]. AB - BACKGROUND: This study was designed to determine if a natural mineral supplement, sierrasil, alone and in combination with a cat's claw extract (Uncaria guianensis), vincaria, has therapeutic potential in mild to moderate osteoarthritis of the knee. METHODS: Patients (n = 107) with mild to moderate osteoarthritis of the knee were randomly assigned to one of 4 groups; high dose sierrasil (3 g/day), low dose sierrasil (2 g/day), low dose sierrasil (2 g/day) + cat's claw extract (100 mg/day) or placebo, administered for 8 weeks. Treatment was double blinded. Primary efficacy variables were WOMAC scores (A, B, C and total). Visual analog score (VAS) for pain, consumption of rescue medication (paracetamol), and tolerability were secondary variables. Safety measures included vital signs and laboratory-based assays. RESULTS: Ninety-one of the 107 patients successfully completed the protocol. All four groups showed improvement in WOMAC and VAS scores after 8 weeks (p < 0.001), in all 3 groups receiving sierrasil the magnitude of benefits were greater vs. placebo (WOMAC Total 38-43% vs. 27%) but this was not statistically significant. In reference to baseline values sierrasil treated groups had a considerably faster onset of benefits. Placebo-treated individuals failed to show significant benefits at 4 weeks (11% reduction in total WOMAC). In contrast, after 1 or 2 weeks of therapy all the sierrasil groups displayed significant reductions in WOMAC scores (p < 0.05) and at week 4 displayed a 38-43% improvement. VAS was significantly improved at 4 weeks in all groups (p < 0.001) but was significantly greater in all sierrasil groups compared to placebo (p < 0.05). Rescue medication use was 28-23% lower in the herbomineral combination and high dose sierrasil groups although not statistically different from placebo (P = 0.101 and P = 0.193, respectively). Tolerability was good for all groups, no serious adverse events were noted and safety parameters remained unchanged. CONCLUSION: The natural mineral supplement, sierrasil alone and in combination with a cat's claw extract, improved joint health and function within 1-2 weeks of treatment but significant benefits over placebo were not sustained, possibly due to rescue medication masking. Sierrasil may offer an alternative therapy in subjects with joint pain and dysfunction. PMID- 16242033 TI - Role of spinal cord glutamate transporter during normal sensory transmission and pathological pain states. AB - Glutamate is a neurotransmitter critical for spinal excitatory synaptic transmission and for generation and maintenance of spinal states of pain hypersensitivity via activation of glutamate receptors. Understanding the regulation of synaptically and non-synaptically released glutamate associated with pathological pain is important in exploring novel molecular mechanisms and developing therapeutic strategies of pathological pain. The glutamate transporter system is the primary mechanism for the inactivation of synaptically released glutamate and the maintenance of glutamate homeostasis. Recent studies demonstrated that spinal glutamate transporter inhibition relieved pathological pain, suggesting that the spinal glutamate transporter might serve as a therapeutic target for treatment of pathological pain. However, the exact function of glutamate transporter in pathological pain is not completely understood. This report will review the evidence for the role of the spinal glutamate transporter during normal sensory transmission and pathological pain conditions and discuss potential mechanisms by which spinal glutamate transporter is involved in pathological pain. PMID- 16242034 TI - Self-prescribing among young Norwegian doctors: a nine-year follow-up study of a nationwide sample. AB - BACKGROUND: Self-prescribing among doctors is common, but no longitudinal studies have documented this issue. We studied the self-prescribing behaviour among young Norwegian physicians and the predictors of self-prescribing. METHODS: We conducted a nationwide, prospective and longitudinal study following young Norwegian physicians from internship through the subsequent nine years using three postal questionnaires. Chi-square tests and logistic regression models were applied. RESULTS: About 54% of the physicians in their fourth and ninth postgraduate years had self-prescribed medication at least once during the previous year. Among those who had used prescription medication during the previous year, about 90% had self-prescribed. Self-prescribing behaviour did not differ significantly between men and women, or according to the type of work at any time. The most frequently self-prescribed medications were antibiotics (71% 81%), contraceptives (24%-25%), analgesics (18%-21%), and hypnotics (9%-12%). Those who had needed treatment for mental problems had self-prescribed hypnotics and sedatives to a greater extent than the others. Being male, having self prescribed during internship, somatic complaints, mental distress, subjective health complaints, and not having sought help from a general practitioner, were significant adjusted predictors of self-prescribing in the ninth postgraduate year. CONCLUSION: The level of self-prescribing among young Norwegian physicians is relatively high, and this behaviour is established early in their professional lives. Although self-prescribing is acceptable in some situations, physicians should seek professional help for illness. Efforts to inculcate more rational help-seeking behaviour should probably start in medical schools. PMID- 16242035 TI - An online survey of chiropractors' opinions of Continuing Education. AB - BACKGROUND: Continuing Education (CE) for chiropractors is mandatory for licensure in most North American jurisdictions. Numerous chiropractic colleges have begun collaborating with universities to offer master's degree programs. Distance education master's degree programs may be desirable to allow full-time practicing doctors to further their post-graduate education. The present survey sought to answer three questions. First, what is the level of satisfaction of chiropractors with their continuing education? Second, what is the level of interest of chiropractors in online master's degree programs? Lastly, what is the response rate of chiropractors to an online survey? METHODS: An online survey consisting of 22 multiple choice questions was e-mailed to 1000 chiropractors randomly selected from the mailing list of an online chiropractic newsletter. Upon completion of the questionnaire, participants' answers were saved on a secure site. Data analysis included evaluation of the demographic characteristics of the respondents, their opinions of and patterns of taking CE including online education, preferred learning formats, and their interest in proposed online master's degree programs. A survey response rate was determined. RESULTS: Nearly 86% of respondents felt their previously completed CE courses were either somewhat or extremely satisfactory. Over ninety percent of respondents who had completed online or distance CE courses found them to be somewhat or extremely satisfactory. Almost half the respondents indicated that they most preferred online distance learning, while 34.08% most preferred face-to-face interaction. Fifty-three percent of respondents indicated an interest in starting a master's degree program; however 70.46% of respondents were interested in an online master's degree program that would offer CE credit. A response rate of 35.8% was obtained. CONCLUSION: Satisfaction among chiropractors with CE programs is high. The notion of completing a part-time online master's degree (or online combined with face-to-face interaction) appears to be popular among respondents, with a M.Sc. in Chiropractic Sciences being the most popular of those mentioned. Online surveys are a viable method of obtaining opinion in a cost and time efficient manner; there are some sources of bias involved in this type of research, and numerous steps need to be taken to obtain a suitable response rate. PMID- 16242036 TI - Sub-chronic inhalation of high concentrations of manganese sulfate induces lower airway pathology in rhesus monkeys. AB - BACKGROUND: Neurotoxicity and pulmonary dysfunction are well-recognized problems associated with prolonged human exposure to high concentrations of airborne manganese. Surprisingly, histological characterization of pulmonary responses induced by manganese remains incomplete. The primary objective of this study was to characterize histologic changes in the monkey respiratory tract following manganese inhalation. METHODS: Subchronic (6 hr/day, 5 days/week) inhalation exposure of young male rhesus monkeys to manganese sulfate was performed. One cohort of monkeys (n = 4-6 animals/exposure concentration) was exposed to air or manganese sulfate at 0.06, 0.3, or 1.5 mg Mn/m3 for 65 exposure days. Another eight monkeys were exposed to manganese sulfate at 1.5 mg Mn/m3 for 65 exposure days and held for 45 or 90 days before evaluation. A second cohort (n = 4 monkeys per time point) was exposed to manganese sulfate at 1.5 mg Mn/m3 and evaluated after 15 or 33 exposure days. Evaluations included measurement of lung manganese concentrations and evaluation of respiratory histologic changes. Tissue manganese concentrations were compared for the exposure and control groups by tests for homogeneity of variance, analysis of variance, followed by Dunnett's multiple comparison. Histopathological findings were evaluated using a Pearson's Chi Square test. RESULTS: Animals exposed to manganese sulfate at > or = 0.3 mg Mn/m3 for 65 days had increased lung manganese concentrations. Exposure to manganese sulfate at 1.5 mg Mn/m3 for > or = 15 exposure days resulted in increased lung manganese concentrations, mild subacute bronchiolitis, alveolar duct inflammation, and proliferation of bronchus-associated lymphoid tissue. Bronchiolitis and alveolar duct inflammatory changes were absent 45 days post exposure, suggesting that these lesions are reversible upon cessation of subchronic high-dose manganese exposure. CONCLUSION: High-dose subchronic manganese sulfate inhalation is associated with increased lung manganese concentrations and small airway inflammatory changes in the absence of observable clinical signs. Subchronic exposure to manganese sulfate at exposure concentrations (< or = 0.3 mg Mn/m3) similar to the current 8-hr occupational threshold limit value established for inhaled manganese was not associated with pulmonary pathology. PMID- 16242037 TI - Substrate specificity analysis of protein kinase complex Dbf2-Mob1 by peptide library and proteome array screening. AB - BACKGROUND: The mitotic exit network (MEN) is a group of proteins that form a signaling cascade that is essential for cells to exit mitosis in Saccharomyces cerevisiae. The MEN has also been implicated in playing a role in cytokinesis. Two components of this signaling pathway are the protein kinase Dbf2 and its binding partner essential for its kinase activity, Mob1. The components of MEN that act upstream of Dbf2-Mob1 have been characterized, but physiological substrates for Dbf2-Mob1 have yet to be identified. RESULTS: Using a combination of peptide library selection, phosphorylation of optimal peptide variants, and screening of a phosphosite array, we found that Dbf2-Mob1 preferentially phosphorylated serine over threonine and required an arginine three residues upstream of the phosphorylated serine in its substrate. This requirement for arginine in peptide substrates could not be substituted with the similarly charged lysine. This specificity determined for peptide substrates was also evident in many of the proteins phosphorylated by Dbf2-Mob1 in a proteome chip analysis. CONCLUSION: We have determined by peptide library selection and phosphosite array screening that the protein kinase Dbf2-Mob1 preferentially phosphorylated substrates that contain an RXXS motif. A subsequent proteome microarray screen revealed proteins that can be phosphorylated by Dbf2-Mob1 in vitro. These proteins are enriched for RXXS motifs, and may include substrates that mediate the function of Dbf2-Mob1 in mitotic exit and cytokinesis. The relatively low degree of sequence restriction at the site of phosphorylation suggests that Dbf2 achieves specificity by docking its substrates at a site that is distinct from the phosphorylation site. PMID- 16242038 TI - A role for airway remodeling during respiratory syncytial virus infection. AB - BACKGROUND: Severe respiratory syncytial virus infection (RSV) during infancy has been shown to be a major risk factor for the development of subsequent wheeze. However, the reasons for this link remain unclear. The objective of this research was to determine the consequences of early exposure to RSV and allergen in the development of subsequent airway hyperreactivity (AHR) using a developmental time point in the mouse that parallels that of the human neonate. METHODS: Weanling mice were sensitized and challenged with ovalbumin (Ova) and/or infected with RSV. Eight days after the last allergen challenge, various pathophysiological endpoints were examined. RESULTS: AHR in response to methacholine was enhanced only in weanling mice exposed to Ova and subsequently infected with RSV. The increase in AHR appeared to be unrelated to pulmonary RSV titer. Total bronchoalveolar lavage cellularity in these mice increased approximately two-fold relative to Ova alone and was attributable to increases in eosinophil and lymphocyte numbers. Enhanced pulmonary pathologies including persistent mucus production and subepithelial fibrosis were observed. Interestingly, these data correlated with transient increases in TNF-alpha, IFN-gamma, IL-5, and IL-2. CONCLUSION: The observed changes in pulmonary structure may provide an explanation for epidemiological data suggesting that early exposure to allergens and RSV have long-term physiological consequences. Furthermore, the data presented here highlight the importance of preventative strategies against RSV infection of atopic individuals during neonatal development. PMID- 16242039 TI - Identification of genes involved in Ca2+ ionophore A23187-mediated apoptosis and demonstration of a high susceptibility for transcriptional repression of cell cycle genes in B lymphoblasts from a patient with Scott syndrome. AB - BACKGROUND: In contrast to other agents able to induce apoptosis of cultured cells, Ca2+ ionophore A23187 was shown to elicit direct activation of intracellular signal(s). The phenotype of the cells derived from patients having the hemorrhagic disease Scott syndrome, is associated with an abnormally high proportion of apoptotic cells, both in basal culture medium and upon addition of low ionophore concentrations in long-term cultures. These features are presumably related to the mutation also responsible for the defective procoagulant plasma membrane remodeling. We analyzed the specific transcriptional re-programming induced by A23187 to get insights into the effect of this agent on gene expression and a defective gene regulation in Scott cells. RESULTS: The changes in gene expression upon 48 hours treatment with 200 nM A23187 were measured in Scott B lymphoblasts compared to B lymphoblasts derived from the patient's daughter or unrelated individuals using Affymetrix microarrays. In a similar manner in all of the B cell lines, results showed up-regulation of 55 genes, out of 12,000 represented sequences, involved in various pathways of the cell metabolism. In contrast, a group of 54 down-regulated genes, coding for histones and proteins involved in the cell cycle progression, was more significantly repressed in Scott B lymphoblasts than in the other cell lines. These data correlated with the alterations of the cell cycle phases in treated cells and suggested that the potent effect of A23187 in Scott B lymphoblasts may be the consequence of the underlying molecular defect. CONCLUSION: The data illustrate that the ionophore A23187 exerts its pro-apoptotic effect by promoting a complex pattern of genetic changes. These results also suggest that a subset of genes participating in various steps of the cell cycle progress can be transcriptionally regulated in a coordinated fashion. Furthermore, this research brings a new insight into the defect in cultured Scott B lymphoblasts, leading to hypothesize that a mutated gene plays a role not only in membrane remodeling but also in signal transduction pathway(s) leading to altered transcriptional regulation of cell cycle genes. PMID- 16242041 TI - Small business owners' health and safety intentions: a cross-sectional survey. AB - BACKGROUND: Little is known about the variables underlying small business owners' behavioural intentions toward workplace health and safety. This project explores the relationship between three mediating variables (Attitude Toward Safety, Subjective Norm and Perceived Behavioural Control) and owners' Intentions Toward Safety, following the Theory of Planned Behaviour. We also investigate the role of beliefs underlying each mediating variable. METHODS: Seven hundred businesses (5-50 employees) were randomly selected from 4084 eligible companies in a manufacturing business database (SIC codes 24 to 39). The 348 respondents are on average 51 yrs of age, 86% male, 96% white and have 2 to 4 years of post secondary school. RESULTS: All three mediator variables are significantly correlated with Intentions Toward Safety; Attitude Toward Safety shows the strongest correlation, which is confirmed by path analysis. Owners with higher attitudes toward safety have a higher probability of believing that improving workplace health and safety will make employees' healthier and happier, show that they care, increase employee productivity, lower workers' compensation costs, increase product quality and lower costs. CONCLUSION: These results suggest that interventions aimed at increasing owners' health and safety intentions (and thus, behaviours) should focus on demonstrating positive employee health and product quality outcomes. PMID- 16242040 TI - Combustion-derived nanoparticles: a review of their toxicology following inhalation exposure. AB - This review considers the molecular toxicology of combustion-derived nanoparticles (CDNP) following inhalation exposure. CDNP originate from a number of sources and in this review we consider diesel soot, welding fume, carbon black and coal fly ash. A substantial literature demonstrates that these pose a hazard to the lungs through their potential to cause oxidative stress, inflammation and cancer; they also have the potential to redistribute to other organs following pulmonary deposition. These different CDNP show considerable heterogeneity in composition and solubility, meaning that oxidative stress may originate from different components depending on the particle under consideration. Key CDNP associated properties of large surface area and the presence of metals and organics all have the potential to produce oxidative stress. CDNP may also exert genotoxic effects, depending on their composition. CDNP and their components also have the potential to translocate to the brain and also the blood, and thereby reach other targets such as the cardiovascular system, spleen and liver. CDNP therefore can be seen as a group of particulate toxins unified by a common mechanism of injury and properties of translocation which have the potential to mediate a range of adverse effects in the lungs and other organs and warrant further research. PMID- 16242043 TI - Geographical variation of cerebrovascular disease in New York State: the correlation with income. AB - BACKGROUND: Income is known to be associated with cerebrovascular disease; however, little is known about the more detailed relationship between cerebrovascular disease and income. We examined the hypothesis that the geographical distribution of cerebrovascular disease in New York State may be predicted by a nonlinear model using income as a surrogate socioeconomic risk factor. RESULTS: We used spatial clustering methods to identify areas with high and low prevalence of cerebrovascular disease at the ZIP code level after smoothing rates and correcting for edge effects; geographic locations of high and low clusters of cerebrovascular disease in New York State were identified with and without income adjustment. To examine effects of income, we calculated the excess number of cases using a non-linear regression with cerebrovascular disease rates taken as the dependent variable and income and income squared taken as independent variables. The resulting regression equation was: excess rate = 32.075-1.22 x 10(-4)(income)+ 8.068x10(-10)(income2), and both income and income squared variables were significant at the 0.01 level. When income was included as a covariate in the non-linear regression, the number and size of clusters of high cerebrovascular disease prevalence decreased. Some 87 ZIP codes exceeded the critical value of the local statistic yielding a relative risk of 1.2. The majority of low cerebrovascular disease prevalence geographic clusters disappeared when the non-linear income effect was included. For linear regression, the excess rate of cerebrovascular disease falls with income; each 10,000 dollars increase in median income of each ZIP code resulted in an average reduction of 3.83 observed cases. The significant nonlinear effect indicates a lessening of this income effect with increasing income. CONCLUSION: Income is a non-linear predictor of excess cerebrovascular disease rates, with both low and high observed cerebrovascular disease rate areas associated with higher income. Income alone explains a significant amount of the geographical variance in cerebrovascular disease across New York State since both high and low clusters of cerebrovascular disease dissipate or disappear with income adjustment. Geographical modeling, including non-linear effects of income, may allow for better identification of other non-traditional risk factors. PMID- 16242042 TI - Targeting energy metabolism in brain cancer: review and hypothesis. AB - Malignant brain tumors are a significant health problem in children and adults and are often unmanageable. As a metabolic disorder involving the dysregulation of glycolysis and respiration, malignant brain cancer is potentially manageable through changes in metabolic environment. A radically different approach to brain cancer management is proposed that combines metabolic control analysis with the evolutionarily conserved capacity of normal cells to survive extreme shifts in physiological environment. In contrast to malignant brain tumors that are largely dependent on glycolysis for energy, normal neurons and glia readily transition to ketone bodies (beta-hydroxybutyrate) for energy in vivo when glucose levels are reduced. The bioenergetic transition from glucose to ketone bodies metabolically targets brain tumors through integrated anti-inflammatory, anti-angiogenic, and pro-apoptotic mechanisms. The approach focuses more on the genomic flexibility of normal cells than on the genomic defects of tumor cells and is supported from recent studies in orthotopic mouse brain tumor models and in human pediatric astrocytoma treated with dietary energy restriction and the ketogenic diet. PMID- 16242044 TI - A method of precise mRNA/DNA homology-based gene structure prediction. AB - BACKGROUND: Accurate and automatic gene finding and structural prediction is a common problem in bioinformatics, and applications need to be capable of handling non-canonical splice sites, micro-exons and partial gene structure predictions that span across several genomic clones. RESULTS: We present a mRNA/DNA homology based gene structure prediction tool, GIGOgene. We use a new affine gap penalty splice-enhanced global alignment algorithm running in linear memory for a high quality annotation of splice sites. Our tool includes a novel algorithm to assemble partial gene structure predictions using interval graphs. GIGOgene exhibited a sensitivity of 99.08% and a specificity of 99.98% on the Genie learning set, and demonstrated a higher quality of gene structural prediction when compared to Sim4, est2genome, Spidey, Galahad and BLAT, including when genes contained micro-exons and non-canonical splice sites. GIGOgene showed an acceptable loss of prediction quality when confronted with a noisy Genie learning set simulating ESTs. CONCLUSION: GIGOgene shows a higher quality of gene structure prediction for mRNA/DNA spliced alignment when compared to other available tools. PMID- 16242045 TI - Silencing of human T-cell leukemia virus type I gene transcription by epigenetic mechanisms. AB - BACKGROUND: Human T-cell leukemia virus type I (HTLV-I) causes adult T-cell leukemia (ATL) after a long latent period. Among accessory genes encoded by HTLV I, the tax gene is thought to play a central role in oncogenesis. However, Tax expression is disrupted by several mechanims including genetic changes of the tax gene, deletion/hypermethylation of 5'-LTR. To clarify the role of epigenetic changes, we analyzed DNA methylation and histone modification in the whole HTLV-I provirus genome. RESULTS: The gag, pol and env genes of HTLV-I provirus were more methylated than pX region, whereas methylation of 5'-LTR was variable and 3'-LTR was not methylated at all. In ATL cell lines, complete DNA methylation of 5'-LTR was associated with transcriptional silencing of viral genes. HTLV-I provirus was more methylated in primary ATL cells than in carrier state, indicating the association with disease progression. In seroconvertors, DNA methylation was already observed in internal sequences of provirus just after seroconversion. Taken together, it is speculated that DNA methylation first occurs in the gag, pol and env regions and then extends in the 5' and 3' directions in vivo, and when 5'-LTR becomes methylated, viral transcription is silenced. Analysis of histone modification in the HTLV-I provirus showed that the methylated provirus was associated with hypoacetylation. However, the tax gene transcript could not be detected in fresh ATL cells regardless of hyperacetylated histone H3 in 5' LTR. The transcription rapidly recovered after in vitro culture in such ATL cells. CONCLUSION: These results showed that epigenetic changes of provirus facilitated ATL cells to evade host immune system by suppressing viral gene transcription. In addition, this study shows the presence of another reversible mechanism that suppresses the tax gene transcription without DNA methylation and hypoacetylated histone. PMID- 16242046 TI - Short- and long-term cause-specific survival of patients with inflammatory breast cancer. AB - BACKGROUND: Inflammatory breast cancer (IBC) had been perceived to have a poor prognosis. Oncologists were not enthusiastic in the past to give aggressive treatment. Single institution studies tend to have small patient numbers and limited years of follow-up. Most studies do not report 10-, 15- or 20-year results. METHODS: Data was obtained from the population-based database of the Surveillance, Epidemiology, and End Results program of the National Cancer Institute from 1975-1995 using SEER*Stat5.0 software. This period of 21 years was divided into 7 periods of 3 years each. The years were chosen so that there was adequate follow-up information to 2000. ICD-O-2 histology 8530/3 was used to define IBC. The lognormal model was used for statistical analysis. RESULTS: A total of 1684 patients were analyzed, of which 84% were white, 11% were African Americans, and 5% belonged to other races. Age distribution was < 30 years in 1%, 30-40 in 11%, 40-50 in 22%, 50-60 in 24%, 60-70 in 21%, and > 70 in 21%. The lognormal model was validated for 1975-77 and for 1978-80, since the 10-, 15- and 20-year cause-specific survival (CSS) rates, could be calculated using the Kaplan Meier method with data available in 2000. The data were then used to estimate the 10-, 15- and 20-year CSS rates for the more recent years, and to study the trend of improvement in survival. There were increasing incidences of IBC: 134 patients in the 1975-77 period to 416 patients in the 1993-95 period. The corresponding 20 year CSS increased from 9% to 20% respectively with standard errors of less than 4%. CONCLUSION: The improvement of survival during the study period may be due to introduction of more aggressive treatments. However, there seem to be no further increase of long-term CSS, which should encourage oncologists to find even more effective treatments. Because of small numbers of patients, randomized studies will be difficult to conduct. The SEER population-based database will yield the best possible estimate of the trend in improvement of survival for patients with IBC. PMID- 16242047 TI - Contrasting phenotypes of putative proprioceptive and nociceptive trigeminal neurons innervating jaw muscle in rat. AB - BACKGROUND: Despite the clinical significance of muscle pain, and the extensive investigation of the properties of muscle afferent fibers, there has been little study of the ion channels on sensory neurons that innervate muscle. In this study, we have fluorescently tagged sensory neurons that innervate the masseter muscle, which is unique because cell bodies for its muscle spindles are in a brainstem nucleus (mesencephalic nucleus of the 5th cranial nerve, MeV) while all its other sensory afferents are in the trigeminal ganglion (TG). We examine the hypothesis that certain molecules proposed to be used selectively by nociceptors fail to express on muscle spindles afferents but appear on other afferents from the same muscle. RESULTS: MeV muscle afferents perfectly fit expectations of cells with a non-nociceptive sensory modality: Opiates failed to inhibit calcium channel currents (I(Ca)) in 90% of MeV neurons, although ICa were inhibited by GABA(B) receptor activation. All MeV afferents had brief (1 msec) action potentials driven solely by tetrodotoxin (TTX)-sensitive Na channels and no MeV afferent expressed either of three ion channels (TRPV1, P2X3, and ASIC3) thought to be transducers for nociceptive stimuli, although they did express other ATP and acid-sensing channels. Trigeminal masseter afferents were much more diverse. Virtually all of them expressed at least one, and often several, of the three putative nociceptive transducer channels, but the mix varied from cell to cell. Calcium currents in 80% of the neurons were measurably inhibited by mu-opioids, but the extent of inhibition varied greatly. Almost all TG masseter afferents expressed some TTX-insensitive sodium currents, but the amount compared to TTX sensitive sodium current varied, as did the duration of action potentials. CONCLUSION: Most masseter muscle afferents that are not muscle spindle afferents express molecules that are considered characteristic of nociceptors, but these putative muscle nociceptors are molecularly diverse. This heterogeneity may reflect the mixture of metabosensitive afferents which can also signal noxious stimuli and purely nociceptive afferents characteristic of muscle. PMID- 16242048 TI - Satisfaction is not all--patients' perceptions of outcome of general practice consultations, a qualitative study. AB - BACKGROUND: Evaluation of outcome in general practice can be seen from different viewpoints. In this study we focus on the concepts patients use to describe the outcome of a consultation with a GP. METHOD: Patients were interviewed within a week after a consultation with a GP. The interviews were made with 20 patients in 5 focus groups and 8 individually. They were analysed with a phenomenographic research approach. RESULTS: From the patient's perspective, the outcome of a consultation is about cure or symptom relief, understanding, confirmation, reassurance, change in self-perception and satisfaction. CONCLUSION: General practice consultations are often more important for patients than generally supposed. Understanding is the most basic concept. PMID- 16242049 TI - Does the Angiotensin-converting enzyme (ACE) gene insertion/deletion polymorphism modify the response to ACE inhibitor therapy?--A systematic review. AB - BACKGROUND: Pharmacogenetic testing to individualize ACE inhibitor therapy remains controversial. We conducted a systematic review to assess the effect modification of the insertion/deletion (I/D) polymorphism of the ACE gene on any outcome in patients treated with ACE inhibitors for cardiovascular and/or renal disease. METHODS: Our systematic review involved searching six electronic databases, then contacting the investigators (and pharmaceutical industry representatives) responsible for the creation of these databases. Two reviewers independently selected relevant randomized, placebo-controlled trials and abstracted from each study details on characteristics and quality. RESULTS: Eleven studies met our inclusion criteria. Despite repeated efforts to contact authors, only four of the eleven studies provided sufficient data to quantify the effect modification by genotypes. We observed a trend towards better response to ACE inhibitors in Caucasian DD carriers compared to II carriers, in terms of blood pressure, proteinuria, glomerular filtration rate, ACE activity and progression to end-stage renal failure. Pooling of the results was inappropriate, due to heterogeneity in ethnicity, clinical domains and outcomes. CONCLUSION: Lack of sufficient genetic data from the reviewed studies precluded drawing any convincing conclusions. Better reporting of genetic data are needed to confirm our preliminary observations concerning better response to ACE inhibitors among Caucasian DD carriers as compared to II carriers. PMID- 16242050 TI - Diagnosis and management of synovial sarcoma. AB - Synovial sarcoma is a unique tumor with substantial promise for biologically targeted therapy. Although it demonstrates moderate chemosensitivity, with approximately 50% response rates to ifosfamide- and doxorubicin-containing regimens, it has a diagnostic translocation and a potentially informative chimeric protein product. Although surgical management remains the cornerstone to effect local control, therapeutic advancements are unlikely to occur by continuing to include advanced cases of synovial sarcomas in trials with other soft tissue sarcomas. Rather, attention should be turned toward prospective molecular targets and development of novel agents to exploit them. Research should be directed at understanding the fusion protein of the X,18 translocation and further validating the role of overexpressed proteins in synovial sarcoma. Meanwhile, carefully designed clinical trials of these agents, with translational correlates, will provide in vivo data to complement the preclinical experience. PMID- 16242051 TI - The potential for molecular therapeutic targets in Ewing's sarcoma. AB - Ewing's sarcoma is an uncompromising tumor of children and young adults. Before the introduction of chemotherapy for Ewing's sarcoma, nearly all patients succumbed to their disease, even with highly aggressive approaches to local control. The realization that most patients have micrometastatic disease at presentation, and the identification of active chemotherapeutic agents for this tumor, have resulted in significant improvements in patient survival. Modern therapy for Ewing's sarcoma combines high-dose chemotherapy for systemic control of disease, with advanced surgical and/or radiation therapeutic approaches for local control. Current therapy remains imperfect. Despite optimal management, the cure rate for localized disease is only approximately 70%, whereas the cure rate for metastatic disease at presentation is less than 30%. Patients who experience long-term disease-free survival are at risk for significant side effects of therapy, including infertility, limb dysfunction, and an increased risk for second malignancies. More effective and less toxic therapies are needed. This report presents an overview of dysregulated molecular pathways in Ewing's sarcoma and highlights the possibility that they may serve as therapeutic targets for the disease. Although a great deal of additional investigation is required before most of these approaches can be assessed in the clinic, we think that these potential new targets offer a great deal of hope for patients with Ewing's sarcoma. PMID- 16242052 TI - Molecular research directions in the management of gastrointestinal stromal tumors. AB - Imatinib mesylate (STI571) is an oral 2-phenylaminopyrimidine derivative that acts as a selective inhibitor against several receptor tyrosine kinases and has been viewed as one of the therapeutic success stories of the 21st century. Imatinib was first shown to inhibit the causative molecular translocation in chronic myelogenous leukemia, BCR-ABL. Because imatinib could also inhibit the activity of KIT, a 145-kD transmembrane glycoprotein, and because gastrointestinal stromal tumors (GISTs), the most common mesenchymal tumors of the digestive tract, are characterized by expression of a gain-of-function mutation in KIT, imatinib was used in therapeutic trials of GISTs beginning in 1999. The initial success has now resulted in more widespread use of imatinib for the treatment of patients with GIST. Molecular genetic studies have shown that most GISTs possess a KIT mutation in exon 9, 11, 13, or 17. Clinically, GIST patients with KIT exon 11 mutations (ie, the juxtamembrane region) are the most prevalent and sensitive to imatinib. In addition to the inhibitory effect on KIT, imatinib also inhibits the activity of mutant platelet-derived growth factor receptor-alpha (PDGFRalpha) found in a subset of GIST. What is becoming evident is that there are patients with GIST who lack mutations in KIT or PDGFRalpha, or possess "imatinib-resistant" mutations (such as exon 17 mutations in KIT and exon 18 mutations in PDGFRalpha). These patients typically do not respond well to imatinib therapy. Therefore, identifying additional genetic factors that contribute to the pathogenesis of GIST, independent of KIT and PDGFRalpha, will be important in developing additional anti-GIST therapies. As one might suspect from previous experiences with antitumor therapies, primary and secondary resistance to imatinib is also becoming a major clinical problem in the treatment of this disease. Therefore, new drugs that can serve as alternative therapies in imatinib-resistant patients with GIST or that can be used in combination with imatinib will be needed. As with most recent efforts to derive novel molecular target therapies to treat cancer, improved therapy of GIST will continue to benefit from advances in the molecular characterization of this disease. PMID- 16242053 TI - Strategies and methodologies for identifying molecular targets in sarcomas and other tumors. AB - Sarcomas are a heterogeneous group of tumors that respond poorly to conventional cytotoxic chemotherapy. Sarcomas possess specific molecular characteristics that exist because of unique somatic mutations, vital growth factor or growth factor receptor overexpression, or are critically dependent on host pathways. Currently these tumors are classified on the basis of their tissue of origin and histologic appearance. Gene expression and proteomic analysis of sarcomas may enable reclassification of these tumors and help predict their biologic behavior and devise common therapeutic strategies within a class. It may not be long before cDNA/protein expression profiling and karyotyping complement the current standard pathological evaluation of a newly diagnosed sarcoma, thereby helping the clinician pick targeted agent(s) relevant to the expression profile. The spectacular success of imatinib in patients with gastrointestinal stromal tumor demonstrates how molecular targeting can fulfill the promise of low toxicity and high response rates. Finding new targets, and learning how to use the current generation of targeting drugs in sarcoma, are urgent challenges. Therapeutic clinical trials with a host of new molecular-targeting agents are underway that will drive new paradigms in sarcoma therapeutics. Patients with refractory disease who currently have no viable therapeutic options may have options that open up with the advent of newer targeted approaches, converting their disease from an acute short-lived one to a chronic process with preservation of quality of life while receiving therapy. PMID- 16242054 TI - The role of radiosurgery in the management of malignant brain tumors. AB - Stereotactic radiosurgery (SRS) provides the means for creating lesions in deep seated areas of the brain inaccessible to invasive surgery, using single high doses of focused ionizing radiation, administered using stereotactic guidance. It is a surgical technique designed to produce a specific radiobiological effect within a sharply defined target region in a single treatment session. Its technical application requires a stereotactic coordinate system, highly accurate patient repositioning (usually fixed), and multiple convergent beams of photon radiation. SRS appears to provide no benefit in the upfront treatment of newly diagnosed malignant gliomas but may be used to effectively palliate small well demarcated volumes of recurrent disease. For selected patients with brain metastases treated with whole-brain radiation therapy (WBRT), the addition of SRS improves median survival. In selected patients with brain metastases, it is also rational to withhold WBRT in favor of radiosurgery alone, with WBRT reserved for salvage without a decrease in median survival time. PMID- 16242055 TI - Esthesioneuroblastoma. AB - Esthesioneuroblastoma is a rare tumor for which the published literature does not provide an evidence-based consensus treatment approach. Multimodality therapy including a combination of surgery and radiation appears to provide the best disease-free and overall survival. Surgical resection should proceed through a craniofacial approach if possible. The role of chemotherapy in the initial treatment paradigm is less clear. Chemotherapy should not be used as single modality therapy for initial treatment but may provide additional benefit when used in combination with radiation and surgery, particularly in advanced-stage disease. Combination chemotherapy should be considered as initial therapy for unresectable tumors and metastatic disease, and as salvage therapy in disease recurrence. PMID- 16242056 TI - Intra-arterial chemotherapy of primary brain tumors. AB - Intra-arterial (IA) chemotherapy is a form of regional delivery to brain tumors, designed to enhance the intra-tumoral concentrations of a given drug, in comparison with the intravenous route. Drugs that are likely to benefit from IA delivery have a rapid systemic clearance and include carmustine and other nitrosoureas, cisplatin, carboplatin, etoposide, and methotrexate. Clinical studies have demonstrated activity of IA chemotherapy approaches for low- and high-grade gliomas, and for cerebral lymphomas. However, a survival benefit for IA drug delivery, in comparison with intravenous administration, has not been proven in phase III trials. The technique is limited by the potential for significant vascular and neurologic toxicity, including visual loss, stroke, and leukoencephalopathy. More recent studies suggest that toxicity can be reduced by the use of carboplatin- and methotrexate-based regimens. Further clinical studies will be needed to determine the appropriate role for IA chemotherapy in the treatment of primary brain tumors. PMID- 16242059 TI - A biographical study of food choice capacity: standards, circumstances, and food management skills. AB - OBJECTIVE: Conceptual understanding of how management of food and eating is linked to life course events and experiences. DESIGN/SETTING: Individual qualitative interviews with adults in upstate New York. PARTICIPANTS: Fourteen men and 11 women with moderate to low incomes. PHENOMENON: Food choice capacity. ANALYSIS: Constant comparative method. RESULTS: A conceptual model of food choice capacity emerged. Food choice capacity represented participants' confidence in meeting their standards for food and eating given their food management skills and circumstances. Standards (expectations for how participants felt they should eat) were based on life course events and experiences. Food management skills (mental and physical talents to keep food costs down and prepare meals) were sources of self-esteem for many participants. Most participants had faced challenging and changing circumstances (income, employment, social support, roles, health conditions). Participants linked strong food management skills with high levels of food choice capacity, except in the case of extreme financial circumstances or the absence of strong standards. IMPLICATIONS FOR RESEARCH AND PRACTICE: Recognizing people's experiences and perspectives in food choice is important. Characterizing food management skills as durable, adaptive resources positions them conceptually for researchers and in a way that practitioners can apply in developing programs for adults. PMID- 16242060 TI - Reported attitudes and beliefs toward soy food consumption of soy consumers versus nonconsumers in natural foods or mainstream grocery stores. AB - OBJECTIVE: To examine the attitudes and beliefs of soy foods consumers (SCs) versus nonconsumers (NCs). DESIGN: Seven focus groups were conducted. SETTING: Mainstream or natural foods grocery stores. PARTICIPANTS: Fifty-three participants, ages 18 to 91 years. VARIABLES MEASURED: Focus groups included discussions on lifestyle practices, beliefs about soy, conversion to soy consumption, and suggestions on how to increase soy consumption. ANALYSIS: Common themes were identified, coded, and compared using NVivo computer software. RESULTS: Barriers to soy consumption included soy's image, a lack of familiarity with how to prepare soy foods, and a perception that soy foods were an inadequate flavor substitute for animal-based products. SCs' conversion to regular consumption was initiated by food intolerances, an increased interest in health, or an adoption of a vegetarian or natural foods lifestyle and was sustained because they enjoyed the flavor. Many participants did not know why soy was considered healthful, whereas others identified it as "heart healthy," a source of protein, and good for women's health. Some SCs had become concerned regarding the controversy surrounding breast cancer and soy consumption. CONCLUSIONS AND IMPLICATIONS: Improving soy's image and educating consumers on its preparation could increase soy consumption. PMID- 16242061 TI - Unraveling the web: an evaluation of the content quality, usability, and readability of nutrition web sites. AB - OBJECTIVE: To determine the content quality, general readability, and usability characteristics of consumer nutrition information on the World Wide Web. DESIGN: Almost 500 Web sites were identified for evaluation through 2 different approaches. Of these, 150 were included for further evaluation. Each site was rated on a 27-item tool covering content quality, readability, and usability. ANALYSIS: Summary statistics, means, ranges, and standard deviation were calculated for each study variable. The statistical significance of differences between item means by search strategy was determined using Student's t tests. RESULTS: Web sites identified using popular search engines scored significantly lower for content quality (P < .0001), were easier to navigate (P < .001), had better overall adherence to usability standards (P < .0001), and had lower reading levels compared with those sites identified using a government Web portal. CONCLUSIONS AND IMPLICATIONS: Nutrition Web sites obtained using popular search engines may be aesthetically appealing and easy to use, but they often provide inaccurate nutrition information. As consumers increasingly turn to the World Wide Web for nutrition advice and education, it is imperative that the needs of diverse user populations be identified and addressed. Future nutrition education research should build on these findings by creating strategies that help users find reliable user-friendly gateways to accurate nutrition information on the Internet. PMID- 16242062 TI - Psychosocial correlates of fruit and vegetable consumption among African American men. AB - OBJECTIVE: To determine the best predictors of fruit and vegetable consumption among African American men age 35 years and older. DESIGN: Data (n = 291) from a 2001 nationally representative mail survey commissioned by the American Cancer Society. PARTICIPANTS: 291 African American men age 35 years and older. MAIN OUTCOME MEASURES: DEPENDENT VARIABLES: (1) total fruits and vegetables without fried potatoes, (2) total fruit with juice, and (3) total vegetables without fried potatoes. Independent variables included 3 blocks of predictors: (1) demographics, (2) a set of psychosocial scales, and (3) intent to change variables based on a theoretical algorithm. ANALYSIS: Linear regression models; analysis of variance for the intent to change group. Alpha = .05. RESULTS: Regression model for total fruits and vegetables, significant psychosocial predictors: social norms, benefits, tangible rewards, and barriers-other. Total fruit with juice: social norms, benefits, tangible rewards. Total vegetables, no fried potatoes: tangible rewards, barriers-other interests. CONCLUSIONS AND IMPLICATIONS: For African American men, fruit consumption appears to be motivated by perceived benefits and standards set by important people in their lives; vegetable consumption is a function of extrinsic rewards and preferences for high calorie, fatty foods. The results suggest that communications to increase fruit and vegetable consumption should be crafted to reflect differences in sources of motivation for eating fruits versus eating vegetables. PMID- 16242063 TI - Monitoring of weight in weight loss programs: a double-edged sword? AB - Body size feedback is commonly provided to clients in weight loss programs. The attention focused on one's current, and presumably undesirable, body size can be motivating for those who want to modify a pattern of unhealthful behaviors. For others, this feedback may result in psychological distress and could lead to outcomes that are, in fact, counter to a weight loss strategy, such as attrition from programs, misreporting of dietary intake, and emotional states associated with binge eating. Dietitians and weight loss professionals should provide weight feedback judiciously, especially to vulnerable clients, and perhaps soften the emphasis on body size altogether. PMID- 16242064 TI - Weighing the evidence: benefits of regular weight monitoring for weight control. AB - We address the assertion that weighing obese patients in weight loss programs can be a source of distress and can lead to unfavorable outcomes. Examination of the relevant literature suggests that there is no evidence that weighing by weight loss participants is a cause of negative mood or of body dissatisfaction. Further, there is little evidence that negative mood states or body dissatisfaction lead to a poor outcome in weight loss programs. To the contrary, a number of studies consistently show that more frequent weighing is associated with better weight loss and maintenance. We offer suggestions for dealing with this issue in clinical practice. PMID- 16242065 TI - Food for health and soul: a curriculum designed to facilitate healthful recipe modifications to family favorites. PMID- 16242066 TI - Ultimate nutrition game for mexican american preschoolers. PMID- 16242071 TI - Patients with high-grade gliomas harboring deletions of chromosomes 9p and 10q benefit from temozolomide treatment. AB - Surgical cure of glioblastomas is virtually impossible and their clinical course is mainly determined by the biologic behavior of the tumor cells and their response to radiation and chemotherapy. We investigated whether response to temozolomide (TMZ) chemotherapy differs in subsets of malignant glioblastomas defined by genetic lesions. Eighty patients with newly diagnosed glioblastoma were analyzed with comparative genomic hybridization and loss of heterozygosity. All patients underwent radical resection. Fifty patients received TMZ after radiotherapy (TMZ group) and 30 patients received radiotherapy alone (RT group). The most common aberrations detected were gains of parts of chromosome 7 and losses of 10q, 9p, or 13q. The spectrum of genetic aberrations did not differ between the TMZ and RT groups. Patients treated with TMZ showed significantly better survival than patients treated with radiotherapy alone (19.5 vs 9.3 months). Genomic deletions on chromosomes 9 and 10 are typical for glioblastoma and associated with poor prognosis. However, patients with these aberrations benefited significantly from TMZ in univariate analysis. In multivariate analysis, this effect was pronounced for 9p deletion and for elderly patients with 10q deletions, respectively. This study demonstrates that molecular genetic and cytogenetic analyses potentially predict responses to chemotherapy in patients with newly diagnosed glioblastomas. PMID- 16242072 TI - A3 adenosine receptors modulate hypoxia-inducible factor-1alpha expression in human A375 melanoma cells. AB - Hypoxia-inducible factor-1 (HIF-1) is a key regulator of genes crucial to many aspects of cancer biology. The purine nucleoside, adenosine, accumulates within many tissues under hypoxic conditions, including that of tumors. Because the levels of both HIF-1 and adenosine are elevated within the hypoxic environment of solid tumors, we investigated whether adenosine may regulate HIF-1. Here we show that, under hypoxic conditions (< 2% O2), adenosine upregulates HIF-1alpha protein expression in a dose-dependent and time-dependent manner, exclusively through the A3 receptor subtype. The response to adenosine was generated at the cell surface because the inhibition of A3 receptor expression, by using small interfering RNA, abolished nucleoside effects. A3 receptor stimulation in hypoxia also increases angiopoietin-2 (Ang-2) protein accumulation through the induction of HIF-1alpha. In particular, we found that A3 receptor stimulation activates p44/p42 and p38 mitogen-activated protein kinases, which are required for A3 induced increase of HIF-1alpha and Ang-2. Collectively, these results suggest a cooperation between hypoxic and adenosine signals that ultimately may lead to the increase in HIF-1-mediated effects in cancer cells. PMID- 16242073 TI - Nanoparticles for the optical imaging of tumor E-selectin. AB - We designed a fluorescent peptide-magnetic nanoparticle conjugate that images E selectin expression in mouse xenograft models of Lewis lung carcinoma (LLC) by fluorescence reflectance imaging. It was synthesized by attaching the E-selectin binding peptide (ESBP; CDSDSDITWDQLWDLMK) to a CLIO(Cy5.5) nanoparticle to yield ESBP-CLIO(Cy5.5). Internalization by activated human umbilical vein endothelial cells (HUVECs) was rapid and mediated by E-selectin, indicated by the lack of uptake of nanoparticles bearing similar numbers of a scrambled peptide (Scram). To demonstrate the specificity of E-selectin targeting to ESBP-CLIO(Cy5.5) in vivo, we coinjected ESBP-CLIO(Cy5.5) and Scram-CLIO(Cy3.5) and demonstrated a high Cy5.5/Cy3.5 fluorescence ratio using the LLC. Histology showed that ESBP CLIO was associated with tumor cells as well as endothelial cells, but fluorescence-activated cell sorter analysis showed a far less expression of E selectin on LLC than on HUVECs. Using immunohistochemistry, we demonstrated E selectin expression in both endothelial cells and cancer cells in human prostate cancer specimens. We conclude that ESBP-CLIO(Cy5.5) is a useful probe for imaging E-selectin associated with the LLC tumor, and that E-selectin is expressed not only on endothelial cells but also on LLC cells and human prostate cancer specimens. PMID- 16242074 TI - The vascular-targeting fusion toxin VEGF121/rGel inhibits the growth of orthotopic human bladder carcinoma tumors. AB - Vascular endothelial growth factor (VEGF) and its receptors (FLT-1 and KDR) are overexpressed by human bladder cancer cells and tumor endothelial cells, respectively. Strategies that target VEGF receptors hold promise as antiangiogenic therapeutic approaches to bladder cancer. A fusion protein of VEGF121 and the plant toxin gelonin (rGel) was constructed, expressed in bacteria, and purified to homogeneity. Cytotoxicity experiments of VEGF121/rGel on the highly metastatic 253J B-V human bladder cancer cell line demonstrated that the VEGF121/rGel does not specifically target these cells, whereas Western blot analysis showed no detectable expression of KDR. Treatment with VEGF121/rGel against orthotopically implanted 253J B-V xenografts in nude mice resulted in a significant suppression of bladder tumor growth (approximately 60% inhibition; P < .05) compared to controls. Immunohistochemistry studies of orthotopic 253J B-V tumors demonstrated that KDR is highly overexpressed in tumor vasculature. Immunofluorescence staining with antibodies to CD-31 (blood vessel endothelium) and rGel demonstrated a dramatic colocalization of the construct on tumor neovasculature. Treated tumors also displayed an increase in terminal deoxynucleotidyl transferase-mediated dUTP-biotin end labeling staining compared to controls. Thus, VEGF121/rGel inhibits the growth of human bladder cancer by cytotoxic effects directed against the tumor vascular supply and has significant potential as a novel antiangiogenic therapeutic against human bladder cancer. PMID- 16242075 TI - Disruption of parallel and converging signaling pathways contributes to the synergistic antitumor effects of simultaneous mTOR and EGFR inhibition in GBM cells. AB - Elevated epidermal growth factor receptor (EGFR) and mammalian target of rapamycin (mTOR) signaling are known to contribute to the malignant properties of glioblastoma multiforme (GBM), which include uncontrolled cell proliferation and evasion of apoptosis. Small molecule inhibitors that target these protein kinases have been evaluated in multiple clinical trials for cancer patients, including those with GBM. Here we have examined the cellular and molecular effects of a combined kinase inhibition of mTOR (rapamycin) and EGFR (EKI-785) in U87 and U251 GBM cells. Simultaneous treatment with rapamycin and EKI-785 results in synergistic antiproliferative as well as proapoptotic effects. At a molecular level, rapamycin alone significantly decreases S6 phosphorylation, whereas EKI 785 alone promotes substantially reduced signal transducer and activator of transcription (STAT3) phosphorylation. Treatment with rapamycin alone also increases Akt phosphorylation on Ser-473, but this effect is blocked by a simultaneous administration of EKI-785. Individually, EKI-785 diminishes while rapamycin promotes the binding of the translation inhibitor eukaryotic initiation factor 4E binding protein (4EBP1) to the eukaryotic translation initiation factor 4E (eIF4E). In spite of these opposing effects, the highest level of 4EBP1-eIF4E binding occurs with the combination of the two inhibitors. These results indicate that the inhibition of EGFR and mTOR has distinct as well as common signaling consequences and provides a molecular rationale for the synergistic antitumor effects of EKI-785 and rapamycin administration. PMID- 16242076 TI - Gastrin exerts pleiotropic effects on human melanoma cell biology. AB - The effects of gastrin (G17) on the growth and migration factors of four human melanoma cell lines (HT-144, C32, G-361, and SKMEL-28) were investigated. The expression patterns of cholecystokinin (CCK)(A), CCK(B), and CCK(C) gastrin receptors were investigated in these cells and in seven clinical samples by means of reverse transcription polymerase chain reaction. Melanoma cells appear to express mRNA for CCK(C) receptors, but not for CCK(A) or CCK(B) receptors. Although gastrin does not significantly modify the growth characteristics of the cell lines under study, it significantly modifies their cell migration characteristics. These modifications occur at adhesion level by modifying the expression levels of alpha(v) and beta3 integrins, at motility level by modifying the organization of the actin cytoskeleton, and at invasion level by modifying the expression levels of matrix metalloproteinase 14. We recently demonstrated the presence of CCK(B) receptors in mouse endothelial cells involved in glioblastoma neoangiogenesis. Chronic in vivo administration of a selective CCK(B) receptor antagonist to mice bearing xenografts of human C32 melanoma cells significantly decreased levels of neoangiogenesis, resulting in considerable delays in the growth of these C32 xenografts. In conclusion, our study identifies the pleiotropic effects of gastrin on melanoma cell biology. PMID- 16242077 TI - Sequential molecular and cellular events during neoplastic progression: a mouse syngeneic ovarian cancer model. AB - Studies performed to identify early events of ovarian cancer and to establish molecular markers to support of early detection and the development of chemopreventive regimens have been hindered by a lack of adequate cell models. Taking advantage of the spontaneous transformation of mouse ovarian surface epithelial (MOSE) cells in culture, we isolated and characterized distinct transitional stages of ovarian cancer as the cells progressed from a premalignant nontumorigenic phenotype to a highly aggressive malignant phenotype. Transitional stages were concurrent with progressive increases in proliferation, anchorage independent growth capacity, in vivo tumor formation, and aneuploidy. During neoplastic progression, our ovarian cancer model underwent distinct remodeling of the actin cytoskeleton and focal adhesion complexes, concomitant with downregulation and/or aberrant subcellular localization of two tumor-suppressor proteins E-cadherin and connexin-43. In addition, we demonstrate that epigenetic silencing of E-cadherin through promoter methylation is associated with neoplastic progression of our ovarian cancer model. These results establish critical interactions between cellular cytoskeletal remodeling and epigenetic silencing events in the progression of ovarian cancer. Thus, our MOSE model provides an excellent tool to identify both cellular and molecular changes in the early and late stages of ovarian cancer, to evaluate their regulation, and to determine their significance in an immunocompetent in vivo environment. PMID- 16242081 TI - Hypersensitivity reactions to vaccine components. AB - Vaccines are responsible for the control of many infectious diseases that were once common in the United States, including polio, measles, diphtheria, pertussis (whooping cough), rubella (German measles), mumps, tetanus, and Haemophilus influenzae type b. National efforts to generate collaboration between federal, state, and local governments and public and private health care providers have resulted in record high levels of vaccination coverage in the United States. The high rate of US vaccinations is paralleled by growing concerns about the safety of their delivery. The variety of substances used in vaccines sometimes causes the development of cutaneous reactions in susceptible adults and children. This article will review adverse cutaneous events consistent with hypersensitivity reactions to the following ingredients in vaccines: aluminum, thimerosal, 2 phenoxyethanol, formaldehyde, and neomycin. PMID- 16242082 TI - Patch-test reactions to thioureas are frequently relevant. AB - BACKGROUND: Thioureas are used as antioxidants in the manufacture of rubber (especially neoprene) and as fixatives in photography and photocopy paper. The number of reported cases of allergic contact dermatitis from thioureas is relatively low, and standard patch test series often do not contain thiourea allergens. Thioureas are included in our institution's standard patch test series. OBJECTIVE: To examine the occurrence and relevance of patch-test reactions to thioureas. METHODS: We retrospectively analyzed patch-test data for patients evaluated between January 1999 and June 2001. Patients included in the study underwent testing with a standard screening series of allergens and a standardized patch testing technique. Our standard patch test series includes mixed dialkyl thioureas at a concentration of 1% in petrolatum. RESULTS: Of 1,368 patients tested with our standard series, 33 patients (2.4%) had patch-test reactions to mixed dialkyl thioureas at day 5. Reactions in 14 (42%) of those patients were of definite clinical relevance, indicating a confirmed exposure to thioureas in many different settings. CONCLUSION: Patch-test reactions to thioureas occur as often as do reactions to many other allergens in the standard series and are often relevant. Institutions should consider including mixed dialkyl thioureas in their standard patch test screening series. PMID- 16242078 TI - Vascular targeting in pancreatic cancer: the novel tubulin-binding agent ZD6126 reveals antitumor activity in primary and metastatic tumor models. AB - ZD6126 is a novel vascular-targeting agent that acts by disrupting the tubulin cytoskeleton of an immature tumor endothelium, leading to an occlusion of tumor blood vessels and a subsequent tumor necrosis. We wanted to evaluate ZD6126 in primary and metastatic tumor models of human pancreatic cancer. Nude mice were injected orthotopically with L3.6pl pancreatic cancer cells. In single and multiple dosing experiments, mice received ZD6126, gemcitabine, a combination of both agents, or no treatment. For the induction of metastatic diseases, additional groups of mice were injected with L3.6pl cells into the spleen. Twenty four hours after a single-dose treatment, ZD6126 therapy led to an extensive central tumor necrosis, which was not seen after gemcitabine treatment. Multiple dosing of ZD6126 resulted in a significant growth inhibition of primary tumors and a marked reduction of spontaneous liver and lymph node metastases. Experimental metastatic diseases could be significantly controlled by a combination of ZD6126 and gemcitabine, as shown by a reduction of the number and size of established liver metastases. As shown by additional in vitro and in vivo experiments, possible mechanisms involve antivascular activities and subsequent antiproliferative and proapoptotic effects of ZD6126 on tumor cells, whereas direct activities against tumor cells seem unlikely. These data highlight the antitumor and antimetastatic effects of ZD6126 in human pancreatic cancer and reveal benefits of adding ZD6126 to standard gemcitabine therapy. PMID- 16242083 TI - Low yield for extended reading of patch tests with topical corticosteroids. AB - BACKGROUND: On the basis of reports that up to 30% of patch test reactions are missed if an extended reading is not performed, we required that patients who were being patch tested with the corticosteroid series return for a reading at least 1 week after placement of the allergens. OBJECTIVE: To report our institutional experience with extended readings (day 7 or beyond) of patch test reactions to the corticosteroid series. METHODS: We retrospectively reviewed patch test reactions to corticosteroids since extended readings were implemented (April 2001 to June 2004). RESULTS: A total of 135 patients were patch tested with 1,656 corticosteroid allergens. On day 5, five patients had five positive patch test reactions; by the time of the extended reading, no new reactions had developed in these patients. Of the patients with no reactions on day 5, two had a positive result at the extended reading: each had a relevant reaction to budesonide 0.1%, one on day 7 and the other on day 9. CONCLUSIONS: Only 2 reactions (to 1,656 corticosteroids) became apparent at the extended reading. Extended readings were of limited value in our experience. PMID- 16242084 TI - Evaluation of the main contact allergens in propolis (1995 to 2005). AB - BACKGROUND: Propolis, the bee glue, is increasingly used in biocosmetics and for the self-treatment of various diseases. OBJECTIVE: Patients reacting to propolis were requested to participate in further testing with the breakdown constituents of the bee glue. METHODS: Twenty-seven patients agreed to be tested with 18 constituents, including four caffeates (the typical allergens of propolis) derived from the sticky exudates of poplar buds. RESULTS: Seven patients did not react to the propolis constituents tested. In the remaining 20 patients, the four caffeates produced strong reactions. Phenylethyl caffeate, which produced positive reactions in 20 patients, was the leading contact allergen. Benzyl caffeate elicited strong responses in 18 patients, and 3-methyl-2-butenyl caffeate produced reactions in 17 patients. Geranyl caffeate produced positive reactions in 11 patients. The flavonoid tectochrysin gave positive results in 2 patients; ferulic acid, coumaric acid, and methyl cinnamate produced weak responses. CONCLUSIONS: In middle Europe, the caffeates are the responsible allergens in propolis allergy. Patients from other countries, where poplar trees do not grow, become allergic to other propolis constituents but not to the caffeates. PMID- 16242085 TI - Chemical analysis of 2,4-toluene diisocyanate, 1,6-hexamethylene diisocyanate and isophorone diisocyanate in petrolatum patch-test preparations. AB - BACKGROUND: Isocyanates with the general formula R-(N=C=O) are theoretically contact sensitizers. However, allergic contact dermatitis (ACD) from isocyanates is seldom reported. In previous reports, patients reacted to their isocyanate based work materials but not to commercial patch-test preparations of isocyanates. Therefore, we suspected that the low frequency of reported ACD from isocyanates was partly due to inadequate commercial preparations. A past study also showed the concentrations of diphenylmethane-4,4'-diisocyanate (4,4'-MDI) in petrolatum preparations to be much lower than declared. OBJECTIVE: In this study, 2,4-toluene diisocyanate (2,4-TDI), 1,6-hexamethylene diisocyanate (1,6-HDI), and isophorone diisocyanate (IPDI) were investigated in a similar fashion. METHODS: In preparations from 12 dermatology departments and two suppliers of patch-test allergens, we determined the isocyanate content as the isocyanate-dibutylamine derivative, using liquid chromatography and mass spectrometry. The preparations were considered stable if the ratio between the stated and found concentrations was within the range of 0.8 to 1.2. RESULTS: Although 28 of 36 investigated preparations had ratios outside of the stable range, they were in its vicinity, which indicates that preparations of 2,4-TDI, 1,6-HDI, and IPDI are more stable than are preparations of 4,4'-MDI where previously reported results showed ratios far outside of stable range. CONCLUSION: As opposed to preparations of 4,4'-MDI, preparations of 2,4-TDI, 1,6-HDI, and IPDI can be considered to be stable. PMID- 16242086 TI - Photoallergic contact sensitization to 6-methylcoumarin in poikiloderma of Civatte. AB - A 49-year-old woman presented with asymptomatic progressive erythema, hyperpigmentation, atrophy, and telangiectasia on her neck. The clinical appearance and histopathology of the skin lesion was consistent with poikiloderma of Civatte. Photopatch-testing with our standard photoallergens yielded a positive reaction to 6-methylcoumarin, which was found in the patient's perfume. This report describes the first case in which photoallergic contact sensitization to 6-methylcoumarin may play an important role in the pathogenesis of poikiloderma of Civatte. PMID- 16242087 TI - Contact allergy to a cocobolo bracelet. AB - Tropical woods are highly valued because of their strength, hardness, and resistance to moisture. These characteristics make them easy to work with and extremely durable, and that is why they have been used in the manufacture of wooden jewelry, musical instruments, furniture, and handles of many different objects. We present a case of a 44-year-old man who developed pruritus, erythema, and blistering around his right wrist, corresponding exactly to the area where he had worn a wooden bracelet. Thin-layer chromatography performed with the extract of the shavings revealed (R)-4-methoxydalbergione and obtusaquinone (the main components of cocobolo wood) and (S)-4'-hydroxy-4-methoxydalbergione (in lower amounts). Patch-testing with sawdust from the bracelet resulted in a very strong reaction. Patch tests with the pure constituents yielded +++ reactions to the main sensitizers of cocobolo, including obtusaquinone, but also to sensitizers present in other rosewoods. This last fact can be explained by cross-reactivity between different dalbergiones. Contact dermatitis from tropical woods is more frequent than thought, owing to their high sensitizing properties. An exhaustive search can identify the allergen responsible in many cases. PMID- 16242088 TI - Allergic contact dermatitis to a urologic lubricant. PMID- 16242089 TI - Painful edema of the cheek. PMID- 16242090 TI - Patch-testing with dust mite (Dermatophagoides mix): still a high rate of reactivity with 0.1% concentration. PMID- 16242091 TI - [Precarious conditions for controlling the HIV epidemic]. PMID- 16242092 TI - [The value of the Letters to the Editor]. PMID- 16242093 TI - [Reporting of newly diagnosed HIV infections in Catalonia, Barcelona, Spain. Implementation and results]. AB - OBJECTIVES: To describe newly diagnosed HIV infections from the HIV Reporting System in Catalonia (2001-2003), and to compare the characteristics of the epidemic based on the use of the HIV Reporting System and the Catalonian AIDS Registry versus those based on the Catalonian AIDS Registry alone. METHODS: Data were collected from newly diagnosed HIV infections and AIDS cases between 2001 and 2003 in Catalonia. RESULTS: Among the newly diagnosed HIV infections (1,765) the most frequent route of HIV infection was heterosexual transmission (46.8%), followed by men who had sex with men (26.7%), and injecting drug use (19.9%). Out of the 1,210 AIDS cases, the most common route of HIV transmission was injecting drug used (42.2%), followed by (heterosexual transmission 34.5%) and MSM (18.0%). Comparison of routes of HIV transmission in the two reporting systems (HIV/AIDS versus AIDS) revealed statistically significant differences. CONCLUSIONS: The HIV/AIDS Reporting System based on reporting of newly diagnosed HIV infections is feasible, since it has been useful in achieving the objectives of epidemiological HIV infection surveillance. It also provides more accurate information than does the AIDS Registry, which can be used to describe recent patterns of HIV transmission. The completeness of the new reporting system may be enhanced by including the diagnosis of HIV infection among the diseases of mandatory notification. PMID- 16242094 TI - [Mortality and socioeconomic deprivation in the census tracts and districts of Barcelona]. AB - OBJECTIVES: To analyze the association between socioeconomic deprivation and mortality in the census tracts of Barcelona and the distinct patterns of this association in the districts. METHODS: We performed an ecological, cross sectional study based on the 1,812 census tracts and the 10 districts of Barcelona, using socioeconomic variables obtained from the 1991 census and mortality data for 1987-1995. A deprivation index was created through component analysis. Descriptive analyses were performed and multivariate Poisson regression models were adjusted. RESULTS: The greater the socioeconomic deprivation in the census tracts, the higher the mortality. The quartile with the greatest deprivation had a relative risk (RR) of mortality of 1.24 (95% confidence interval [CI], 1.22-1.27) in males and an RR of 1.05 (95% CI, 1.02-1.07) in females (compared with the quartile of census tracts with the lowest deprivation). This association varied according to district: in the old inner city district (district 1), which had the highest mortality and the greatest deprivation, the RR was 1.57 (95% CI, 1.21-2.05) for males, while in district 8, which also has considerable deprivation, mortality was lower. CONCLUSIONS: Analysis of socioeconomic inequalities in mortality in census tracts in an urban area highlights special areas of risk not observed in analysis of districts. PMID- 16242095 TI - Clustering of behavior-related risk factors and its association with subjective health. AB - OBJECTIVES: To describe the clustering of behavior-related risk factors in the adult population of the Autonomous Community of Madrid (Spain) and evaluate the association between the level of aggregation of these factors and suboptimal subjective health. METHODS: Data were drawn from the Non-communicable Disease Risk-Factor Surveillance System (Sistema de Vigilancia de Factores de Riesgo asociados a Enfermedades No Transmisibles [SIVFRENT]). We studied the associations between smoking, high-risk alcohol consumption, leisure-time sedentariness and unbalanced diet in 16,043 persons aged 18-64 years and compared the observed against the expected proportions. Logistic regression was used to estimate the association between clustering of risk factors and suboptimal health (fair, poor and very poor). RESULTS: Almost 20% of subjects had 3 or 4 risk factors simultaneously. Most combinations of 3 risk factors exceeded expectations and, in particular, 4 factor clustering yielded observed/expected quotients of 2.15 (95% confidence interval [CI]: 1.93-2.38) in men and 2.96 (95% CI, 2.46 3.46) in women. In both sexes, the individual factor most closely associated with the remaining risk factors was smoking. Aggregation of risk factors was more frequent among men, younger age groups and subjects with low educational level. Compared with persons with none of the 4 risk factors, those that simultaneously had 3 or 4 more frequently reported suboptimal subjective health (OR = 2.49; 95% CI, 1.59-3.90 in men and OR = 1.96; 95% CI, 1.29-2.97 in women). CONCLUSIONS: Behavior-related risk factors tend to aggregate, and this accumulation is higher among men, younger age groups, and subjects with a low educational level. A greater level of clustering is associated with a higher frequency of suboptimal perceived health. PMID- 16242096 TI - [Prioritization of patients on the waiting list for hip and knee replacement: the patients' views]. AB - OBJECTIVE: To identify patients' views on the criteria used to prioritize patients on the waiting list for total hip or knee prostheses. This study is part of a wider project whose objective is to design an instrument to prioritize patients on the waiting list for both procedures. MATERIAL AND METHODS: We performed a descriptive study with qualitative methodology that provides valuable information on how to improve various aspects of clinical practice and detect solutions that could be useful in decision making. There were 4 focus groups; 2 before the design of the instrument and 2 after. These focus group were composed of patients on the waiting list for knee or hip replacement. RESULTS: Thirty-one patients agreed to participate in the focus groups. All the patients were dissatisfied with the current waiting lists. The most salient factors discussed in meetings were pain, functional capability, and the repercussions of these on the patient's social role. CONCLUSIONS: Although the instrument is designed to be used by health professionals, patients' participation in its design and evaluation allows them to feel more involved in the healthcare process and provides information that more accurately reflects their experiences. The use of this information by health professionals will improve resource optimization and the response to patient needs. PMID- 16242097 TI - [Evaluation of a worksite smoking cessation program]. AB - AIMS: To evaluate the effects of a worksite smoking cessation program conducted to reduce the prevalence of smokers among staff of the town council of Sabadell (Spain). POPULATION AND METHODS: A pre-post intervention study was conducted. A survey was performed to determine the prevalence and characteristics of smokers and to invite them to participate in a smoking cessation program. The intervention involved an individual visit with medical history, measurement of carbon monoxide in expired air and prescription of nicotine replacement therapy followed by 12 sessions of group therapy over 1 year to reinforce abstinence. Follow-up to evaluate abstinence took place at 6, 12 and 24 months. A post intervention survey was carried out to evaluate the program's effectiveness. RESULTS: A total of 1,203 questionnaires were sent with a response rate of 51%. Thirty-three percent of the subjects (n = 204) were smokers and 101 smokers enrolled in the program. Abstinence rates at 12 and 24 months were 45% and 33% respectively with no differences between sexes. Low and moderate nicotine dependence was associated with a greater probability of abstinence at 6 (OR = 6.7; p = 0.01) and 12 months (OR = 5.4: p = 0.05), adjusted by sex, age, and compliance with nicotine replacement therapy. The prevalence of smokers in the post-intervention questionnaire was 19%. In addition, 26% (n = 10) of subjects who quit smoking in the 12-month study period without participating in the program reported quitting because of the program. CONCLUSIONS: The results indicate that worksite smoking cessation programs can substantially contribute to reducing the prevalence of smoking. PMID- 16242098 TI - [Considerations on surveillance of HIV infections. The experience of Navarre (Spain)]. AB - OBJECTIVE AND METHODS: The number and proportion of cases in the HIV registry of Navarre (Spain) that were residents of this region were quantified, according to the census and the healthcare card database. RESULTS: Of the 2,385 persons diagnosed with HIV infection to 2003, only 1,610 (67.5%) were residents of Navarre. The rate of HIV cases diagnosed among residents in Navarre was over 90 cases per million between 1994 and 1998, with values similar to those of Switzerland. In contrast with the time-trend in other European countries, the time-trend in Navarre decreased from 1994 to 2003. Since 2000 the rate of new HIV cases in Navarre has been lower than rates in Portugal, Switzerland, Luxembourg, Belgium, Ireland, and the United Kingdom. CONCLUSION: To prevent overestimation of the number of HIV infections, cases duplicated between regions should be excluded. This could be achieved by a national HIV surveillance system. PMID- 16242099 TI - [Some practical features of clinical practice guidelines]. PMID- 16242100 TI - [Selective serotonin reuptake inhibitors in childhood depression: a soap opera that reflects serious problems of drug safety]. PMID- 16242101 TI - [Comment. The role of regulatory agencies in the evaluation of drug safety: speaking about of selective serotonin receptor inhibitors]. PMID- 16242103 TI - [Journalistic coverage of the violence against women]. PMID- 16242106 TI - [Alan Harold Williams. Pioneer and promoter of health economics]. PMID- 16242107 TI - [Don Quixote and prevention's bad reputation]. PMID- 16242108 TI - Incidence and risk factors of severe traumatic brain injury resulting from road accidents: a population-based study. AB - A population-based study was carried out in 1996-2001 to provide the incidence and to identify the risk factors of severe traumatic brain injury (TBI) resulting from a road accident. The severe TBI was defined as an injury to the brain or the skull, excluding scalp injuries, with an abbreviated injury scale (AIS) severity score greater than 2. The severe TBI of 1238 patients were described. The annual incidence and mortality of severe TBI were, respectively, 13.7 per 100,000 and 5.3 per 100,000. The fatality rate increased from 20% in childhood to 71% over 75 year-old. Compared to restrained car occupants, the odds ratio for having a severe TBI was 18.1 (95% confidence interval, CI=12.8-25.5) for un-helmeted motorcyclists, 9.2 (95% CI=7.5-11.3) for pedestrians, 6.4 (95% CI=4.7-8.8) for un helmeted cyclists, 3.9 (95% CI=3.1-4.8) for unrestrained car occupants and 2.8 (95% CI=2.2-3.5) for helmeted motorcyclists. Even after adjustment for several severity factors, male gender and age above 55 were both risk factors. Prevention programs aiming at improving the head protection should be promoted. The circumstances of the accident should be taken into account to predict a severe TBI. PMID- 16242109 TI - Preparation and chemiresistive properties of nanostructured materials. PMID- 16242110 TI - Nanoclusters in polymer matrices prepared by co-deposition from a gas phase. AB - This paper reviews the fabrication of organic and metal nanoclusters in polymer matrices by three co-deposition techniques. In particular, the structure and properties of polytetrafluoroethylene (PTFE), polychlortrifluoroethylene (PCTFE), polyparaphenylene sulphide (PPS), polystyrene (PS) and polyparaxylylene (PPX) films, containing gold (Au) and dye clusters are discussed. For the first time, dye-filled polymers and multi-component films, consisting of both Au nanoparticles and dye molecules, dispersed in the PTFE matrix were studied. A low temperature plasma was used for film structure modification. Cluster formation process was studied using optical spectroscopy in situ. Transmission electron microscopy (TEM), atomic force microscopy (AFM) and ellipsometry were used for characterisation of the grown films. During Au-PTFE film growth plasmon band shifted from 460-480 nm to 560 nm. Au cluster diameter was in the 3-7 nm range. Plasma treatment of the vapours led to formation of smaller, but more aggregated clusters. During Au-PPS film deposition a two-step growth mechanism was discovered. At the beginning of film growth the plasmon band at 540 nm appeared, but as thickness increased, the band at 430 nm dominated. Without plasma treatment a disordered mixture was deposited, while with plasma treatment large Au aggregates confined with PPS matrix having plasmon band at 620 nm were formed. Dye cluster formation depends on the dye ability to aggregate, its concentration and the properties of the polymer matrix. But cluster formation can also be tuned by varying the deposition conditions. Laser beam evaporation promoted cluster formation, while plasma treatment and dilution in a polymer matrix prevented cluster formation. In all cases both equilibrium and non-equilibrium film structure can be formed using kinetic factor. Asymmetric molecules with bulky substituents were oriented in polymer matrices by applying an electric field in situ or by corona poling. These molecules did not aggregate even at high dye load. The films exhibited second harmonic generation, which demonstrated chromophore orientation in the polymer matrices. PMID- 16242111 TI - Micropatterning of biomolecules on glass surfaces modified with various functional groups using photoactivatable biotin. AB - Biomolecule patterning by photolithographic methods has considerable advantages because a large number of different biomolecules can be assembled on a spatial area by a combinatorial method and complex biomolecule patterning can be created in situ in closed environments such as microfluidic channels. Here, a photobiotin was used as the photoactivatable reagent to create patterned arrays of biomolecules. The variability of photobiotin deposition on glass substrates modified with a variety of materials having carboxyl, lysine, aldehyde, amine groups, and BSA (bovine serum albumin) was characterized by subsequent derivatization with Cy3-labeled streptavidin. The fluorescence images of the photobiotin patterned glass surfaces showed that the BSA/aldehyde-coated glass could be considered as the most appropriate substrate to immobilize photobiotin, in view of the homogeneous immobilization of biomolecules with high density in defined regions and the reduction of nonspecific binding to the surface. In streptavidin equilibrium adsorption assays, the maximum amount of streptavidin Cy3 bound to the BSA/aldehyde-coated glass surface continued to rise with increasing streptavidin-Cy3 concentration until 12.0 microg/mL was reached and the surface then became saturated. Also, a line array of biotin-labeled single strand probe DNAs was created on the BSA/aldehyde-coated glass by photolysis of photobiotin through a slit-type mask and biotin/streptavidin/biotin chemistry, extended to a quantitative measurement of the concentrations of target DNA. The results of target DNA analysis showed linearity over a wide range from 0.5 ng/mL to 5 microg/mL and were reproducible. PMID- 16242112 TI - Bioassay-based screening of microorganisms that degrade dioxin using substrate immobilized microtubes. AB - In the current study, we attempted to develop a method for bioassay-based screening of microorganisms that degrade dioxin. However, a crucial problem encountered was that the standard dioxin 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) added to bacterial medium immediately disappeared from the liquid phase due to its adsorption onto polypropylene (PP) tubes. Among other aromatic hydrocarbons, adsorption onto PP tubes was also observed in beta-naphthoflavone but not in benzo[a]pyrene. Adsorption of TCDD was observed not only onto PP tubes but also onto polystyrene, glass, and PP tubes with low affinity for DNA or protein. Silanization was not effective at preventing adsorption of TCDD. TCDD immobilized onto PP tubes was recovered by organic solvents, including ethanol, methanol, and dimethyl sulfoxide (DMSO). The elution efficiency of the immobilized TCDD by DMSO was approximately 85%. Based on these findings, screening of bacteria that degrade dioxin was attempted as follows. First, TCDD was immobilized onto PP tubes. Second, bacterial suspension was added to the tubes and incubated for biodegradation of TCDD. Third, remaining, immobilized TCDD was eluted by DMSO and subjected to a reporter bioassay to evaluate the level of TCDD. Using this method, we demonstrated successful screening of bacteria that have the potential for degradation of dioxin. PMID- 16242113 TI - [Open minimally invasive parathyroid and thyroid surgery]. AB - Open minimally invasive parathyroidectomy or thyroidectomy (small-incision technique) are frequently performed. Benefits and disadvantages of this approach are discussed in this review. Preoperative patients selection is mandatory and is also discussed. PMID- 16242114 TI - Analysis of the kinetics of the membrane potential generated by cytochrome c oxidase upon single electron injection. AB - In a recent work from this group (Popovic, D. M.; Stuchebrukhov A. A. FEBS Lett. 2004, 566, 126), a model of proton pumping by cytochrome c oxidase (CcO) was proposed. The key element of the model is His291 (bovine notation), a histidine ligand to enzyme's CuB redox center, which plays the role of the pump element. The model assumes that upon electron transfer between heme a and the binuclear catalytic center of the enzyme, two sequential proton transfers occur: First, a proton from Glu242 is transferred to an unprotonated His291, then a second proton, after reprotonation of Glu242 from the negative side of the membrane, is transferred to a hydroxyl group in the binuclear center, a water molecule is formed, and the first proton, due to proton-proton repulsion, is expelled from His291 to the positive side of the membrane, resulting in a pumping event. In the process the free energy of water formation (i.e., reduction of oxygen) is transformed into a proton gradient across the membrane. The model possesses specific kinetic features. It assumes, for example, that upon electron transfer the first proton is transferred to the proton-loading site of the pump, His291, and not to the catalytic center of the enzyme. Here, we analyze the kinetic properties of the proposed model, and calculate the time dependence of the membrane potential generated by CcO upon a single electron injection into the enzyme. These data are directly compared with recent experimental measurements of the membrane potential generated by CcO. Specifically, F to O, and O to E transitions will be discussed. Several enzymes from different organisms (bovine, two bacterial enzymes, and several mutants) are compared and discussed in detail. The kinetic description, however, is phenomenological, and does not include explicitly the nature of the groups involved in proton translocation, except in terms of their position depth within the membrane; thus, the kinetic equations developed here are in fact describe a generic model, similar, e.g., to that proposed earlier by Peter Rich (P.R. Rich, Towards an understanding of the chemistry of oxygen reduction and proton translocation in the iron-copper respiratory oxidases. Aust. J. Plant Physiol. 22 (1995) 479-486), and which is based on the idea of displacement of the pumped protons by the chemical ones. PMID- 16242115 TI - Trehalose loading through the mitochondrial permeability transition pore enhances desiccation tolerance in rat liver mitochondria. AB - Trehalose has extensively been used to improve the desiccation tolerance of mammalian cells. To test whether trehalose improves desiccation tolerance of mammalian mitochondria, we introduced trehalose into the matrix of isolated rat liver mitochondria by reversibly permeabilizing the inner membrane using the mitochondrial permeability transition pore (MPTP). Measurement of the trehalose concentration inside mitochondria using high performance liquid chromatography showed that the sugar permeated rapidly into the matrix upon opening the MPTP. The concentration of intra-matrix trehalose reached 0.29 mmol/mg protein (approximately 190 mM) in 5 min. Mitochondria, with and without trehalose loaded into the matrix, were desiccated in a buffer containing 0.25 M trehalose by diffusive drying. After re-hydration, the inner membrane integrity was assessed by measurement of mitochondrial membrane potential with the fluorescent probe JC 1. The results showed that following drying to similar water contents, the mitochondria loaded with trehalose had significantly higher inner membrane integrity than those without trehalose loading. These findings suggest the presence of trehalose in the mitochondrial matrix affords improved desiccation tolerance to the isolated mitochondria. PMID- 16242117 TI - Anthrax lethal toxin enhances cytokine-induced VCAM-1 expression on human endothelial cells. AB - Vascular endothelial dysfunction is thought to play a prominent role in systemic anthrax pathogenesis. We examined the effect of anthrax lethal toxin (LTx), a key virulence factor of Bacillus anthracis, on the expression of vascular cell adhesion molecule-1 (VCAM-1) on normal and cytokine-stimulated human lung microvascular endothelial cells. Confluent endothelial monolayers were treated with lethal factor (LF), protective antigen (PA), or both (LTx) in the presence or absence of tumor necrosis factor-alpha (TNFalpha). LTx enhanced cytokine induced VCAM-1 expression and monocyte adhesion. LTx alone had no effect on VCAM 1 expression. LF, PA or the combination of a catalytically inactive mutant LF and PA failed to enhance cytokine-induced VCAM-1 expression. Treatment with inhibitors of mitogen-activated protein kinase kinases (MEKs) and mitogen activated protein kinases did not reproduce the VCAM-1 enhancement effect of LTx, a known MEK metalloprotease, suggesting LTx-mediated MEK cleavage may not be a contributing factor. PMID- 16242116 TI - Effects of gramicidin-A on the adsorption of phospholipids to the air-water interface. AB - Prior studies suggest that the hydrophobic surfactant proteins, SP-B and SP-C, promote adsorption of the lipids in pulmonary surfactant to an air-water interface by stabilizing a negatively curved rate-limiting structure that is intermediate between bilayer vesicles and the surface film. This model predicts that other peptides capable of stabilizing negative curvature should also promote lipid adsorption. Previous reports have shown that under appropriate conditions, gramicidin-A (GrA) induces dioleoyl phosphatidylcholine (DOPC), but not dimyristoyl phosphatidylcholine (DMPC), to form the negatively curved hexagonal II (H(II)) phase. The studies reported here determined if GrA would produce the same effects on adsorption of DMPC and DOPC that the hydrophobic surfactant proteins have on the surfactant lipids. Small angle X-ray scattering and (31)P nuclear magnetic resonance confirmed that at the particular conditions used to study adsorption, GrA induced DOPC to form the H(II) phase, but DMPC remained lamellar. Measurements of surface tension showed that GrA in vesicles produced a general increase in the rate of adsorption for both phospholipids. When restricted to the interface, however, in preexisting films, GrA with DOPC, but not with DMPC, replicated the ability of the surfactant proteins to promote adsorption of vesicles containing only the lipids. The correlation between the structural and functional effects of GrA with the two phospholipids, and the similar effects on adsorption of GrA with DOPC and the hydrophobic surfactant proteins with the surfactant lipids fit with the model in which SP-B and SP-C facilitate adsorption by stabilizing a rate-limiting intermediate with negative curvature. PMID- 16242118 TI - Excitatory amino acid transporters expressed by synovial fibroblasts in rats with collagen-induced arthritis. AB - Although previous studies have demonstrated increased levels of the brain neurotransmitter glutamate (Glu) in the synovial fluid from patients with arthritis, not much attention has been paid to the possible role of Glu in joint synovial tissues to date. Constitutive expression of mRNA was for the first time shown with glutamate aspartate transporter, glutamate transporter-1 and excitatory amino acid carrier-1 (EAAC1), in addition to with particular ionotropic and metabotropic Glu receptors, in cultured synovial fibroblasts prepared from knee joints of male Lewis rats. Immunohistochemical analysis revealed high localization of immunoreactive EAAC1 at synovial tissues. The accumulation of [3H]Glu occurred in a temperature- and sodium-dependent manner in cultured synovial fibroblasts, with a Km of 23.1+/-1.1 microM and a Vmax of 237.1+/-31.1 pmol/(mg protein min), respectively. In rats with arthritis induced by immunization to type-II collagen, marked increases were seen in hind paw volume, cytokine mRNA expression and Glu levels in synovial tissues, in addition to histological erosion. In cultured synovial fibroblasts prepared from these arthritic rats, [3H]Glu accumulation was drastically increased with biochemical and pharmacological profiles similar to those seen in normal synovial fibroblasts. The exposure to Glu at 500 microM doubled the incorporation of 5 bromo-2'-deoxyuridine in cultured synovial fibroblasts of arthritic but not normal rats, without significantly affecting mRNA expression of different cytokines in both synovial fibroblasts. These results suggest that Glu may at least in part play a role in mechanisms associated with cellular proliferation through particular transporters functionally expressed by synovium in rheumatoid arthritis. PMID- 16242119 TI - Regarding "antibiotic prophylaxis with azithromycin or penicillin for childhood onset neuropsychiatric disorders". PMID- 16242121 TI - Regarding "antibiotic prophylaxis with azithromycin or penicillin for childhood onset neuropsychiatric disorders". PMID- 16242122 TI - Carbon monoxide and nitric oxide protect against tumor necrosis factor-alpha induced apoptosis in osteoblasts: HO-1 is necessary to mediate the protection. AB - BACKGROUND: Carbon monoxide (CO) and nitric oxide (NO) each have unique roles for various inflammatory states, including inflammatory bone resorption. Although it is known that NO can induce the expression of the cytoprotective enzyme HO-1, there is no information as to whether the protective effect of CO requires NO production or whether CO must induce the expression of HO-1 to exert its functional effects. METHODS: Murine osteoblast cells, MC3T3E1 osteoblasts, were cultured for CO and NO-associated HO-1 experiments and were transfected with pcDNA 3, pcDNA 3-HO-1, control siRNA or HO-1 siRNA using Nucleofector. For cell death measurement, MTT and annexin V assays were used. We performed Western blotting to check the expressions of HO-1 and iNOs and measured the HO-1 enzyme activity. We also measured the amounts of nitrite and nitrate using Griess reagents. RESULTS: The increased expression of HO-1 is required for the protective effect of NO and a single treatment of CO can increase the expression of HO-1, and this is also important for the protective effect of CO in MC3T3E1 osteoblasts. CO as well as NO attenuates the TNF-alpha-induced apoptosis in osteoblasts. The anti-apoptotic effect of CO or NO is not mediated by cGMP, and CO has no effect on the release of NO. The inhibition of HO-1 with using the HO-1 inhibitor ZnPP or HO-1 siRNA resulted in a striking increase of apoptosis in the CO/TNF-alpha-treated cells. Furthermore, HO-1 overexpression showed resistance against the TNF-alpha-induced cytotoxicity in the MC3T3E1 osteoblasts. CONCLUSIONS: There is a need for HO-1 expression to mediate the protection provided by exogenous CO or NO in osteoblasts. PMID- 16242123 TI - Modified and improved method for the measurement of plasma acylcarnitines by online extraction coupled to liquid chromatography-tandem mass spectrometry without derivatization. PMID- 16242124 TI - Multiple actions of U-37883A, an ATP-sensitive K+ channel blocker, on membrane currents in pig urethra. AB - The effects of U-37883A, a vascular ATP-sensitive K(+) channel (K(ATP) channel) blocker, on membrane currents were investigated in pig urethral myocytes by use of patch-clamp techniques (conventional whole-cell recordings, nystatin perforated patches and cell-attached configuration). Tension measurement was also performed to study the effects of U-37883A on the levcromakalim-induced urethral relaxation and the urethral resting tone in the absence and presence of Bay K 8644. Although cumulative application of U-37883A produced a concentration dependent inhibitory effect on the levcromakalim-induced urethral relaxation, U 37883A did not abolish the relaxation. In nystatin-perforated patch recording, K(ATP) currents activated by levcromakalim were inhibited by U-37883A in a concentration-dependent manner (K(i), 4.7 microM). Approximately 10% of the K(ATP) currents still remained even in the presence of 300 microM U-37883A. In cell-attached mode, extracellular application of U-37883A (100 microM) irreversibly inhibited the activity of the levcromakalim-induced K(ATP) channels. In whole-cell configuration, U-37883A suppressed the peak amplitude of voltage dependent Ba(2+) currents in a concentration- and voltage-dependent manner, and at 30 microM, shifted the steady-state inactivation curve of the Ba(2+) currents to the left at -90 mV. These results demonstrate that U-37883A reduces not only the activities of K(ATP) channels but also voltage-dependent Ca(2+) channels. Therefore, it is not appropriate to define U-37883A as solely a vascular K(ATP) channel blocker. PMID- 16242125 TI - Non-neuronal induction of immunoproteasome subunits in an ALS model: possible mediation by cytokines. AB - Protein aggregation is a pathologic hallmark of familial amyotrophic lateral sclerosis caused by mutations in the Cu, Zn superoxide dismutase gene. Although SOD1-positive aggregates can be cleared by proteasomes, aggregates have been hypothesized to interfere with proteasome activity, leading to a vicious cycle that further enhances aggregate accumulation. To address this issue, we measured proteasome activity in transgenic mice expressing a G93A SOD1 mutation. We find that proteasome activity is induced in the spinal cord of such mice compared to controls but is not altered in uninvolved organs such as liver or spleen. This induction within spinal cord is not related to an overall increase in the total number of proteasome subunits, as evidenced by the steady expression levels of constitutive alpha7 and beta5 subunits. In contrast, we found a marked increase of inducible beta proteasome subunits, LMP2, MECL-1 and LMP7. This induction of immunoproteasome subunits does not occur in all spinal cord cell types but appears limited to astrocytes and microglia. The induction of immunoproteasome subunits in G93A spinal cord organotypic slices treated with TNF-alpha and interferon-gamma suggest that certain cytokines may mediate such responses in vivo. Our results indicate that there is an overall increase in proteasome function in the spinal cords of G93A SOD1 mice that correlates with an induction of immunoproteasomes subunits and a shift toward immunoproteasome composition. These results suggest that increased, rather than decreased, proteasome function is a response of certain cell types to mutant SOD1-induced disease within spinal cord. PMID- 16242126 TI - Sertoli cells improve survival of motor neurons in SOD1 transgenic mice, a model of amyotrophic lateral sclerosis. AB - Cell replacement therapy has been widely suggested as a treatment for multiple diseases including motor neuron disease. A variety of donor cells have been tested for treatment including isolated preparations from bone marrow and embryonic spinal cord. Another cell source, Sertoli cells, have been successfully used in models of diabetes, Parkinson's disease and Huntington's disease. The ability of these cells to secrete cytoprotective proteins and their role as 'nurse cells' supporting the function of other cell types in the testes suggest their potential use as neuroprotective cells. The current study examines the ability of Sertoli cells injected into the parenchyma of the spinal cord to protect motor neurons in a mouse model for amyotrophic lateral sclerosis. Seventy transgenic mice expressing the mutant (G93A) human Cu-Zn superoxide dismutase (SOD1) received a unilateral spinal injection of Sertoli-enriched testicular cells into the L4-L5 ventral horn (1 x 10(5) cells total) prior to the onset of clinical symptoms. The animals were euthanized at the end stage of the disease. Histological and morphometric analyses of the transplant site were performed. A significant increase in the number of surviving ChAT positive motor neurons was found ipsilateral to the injection compared with contralateral and uninjected spinal cord. The ipsilateral increase in motor neuron density was dependent upon proximity to the injection site. Sections rostral or caudal to the injection site did not display a similar difference in motor neuron density. Implantation of a Sertoli-cell-enriched preparation has a significant neuroprotective benefit to vulnerable motor neurons in the SOD1 transgenic model. The therapeutic benefit may be the result of secreted neurotrophic factors present at a critical stage of motor neuron degeneration in this model. PMID- 16242127 TI - Inhibition of mitochondrial aldehyde dehydrogenase by nitric oxide-mediated S nitrosylation. AB - Mitochondrial aldehyde dehydrogenase (ALDH2) is responsible for the metabolism of acetaldehyde and other toxic lipid aldehydes. Despite many reports about the inhibition of ALDH2 by toxic chemicals, it is unknown whether nitric oxide (NO) can alter the ALDH2 activity in intact cells or in vivo animals. The aim of this study was to investigate the effects of NO on ALDH2 activity in H4IIE-C3 rat hepatoma cells. NO donors such as S-nitrosoglutathione (GSNO), S-nitroso-N acetylpenicillamine, and 3-morpholinosydnonimine significantly increased the nitrite concentration while they inhibited the ALDH2 activity. Addition of GSH ethylester (GSH-EE) completely blocked the GSNO-mediated ALDH2 inhibition and increased nitrite concentration. To directly demonstrate the NO-mediated S nitrosylation and inactivation, ALDH2 was immunopurified from control or GSNO treated cells and subjected to immunoblot analysis. The anti-nitrosocysteine antibody recognized the immunopurified ALDH2 only from the GSNO-treated samples. All these results indicate that S-nitrosylation of ALDH2 in intact cells leads to reversible inhibition of ALDH2 activity. PMID- 16242128 TI - Developmental shift in the apostat: comparison of neurones and astrocytes. AB - The intrinsic pathway of apoptosis was investigated in cell-free extracts of neurones and astrocytes at various stages of maturation. Neuronal extracts were activated 65-fold after 3 days, 9-fold after 7 days, and were not activated after 10 days in culture. In contrast, astrocyte extracts were activated to a similar extent at all stages, up to 60 days in culture. The co-incubation of neuronal and astrocyte extracts followed by addition of cytochrome c/2'-deoxyadenosine 5' triphosphate led to a 40-fold activation, suggesting that the development associated neuronal shift does not involve the appearance of a dominant inhibitor, but rather downregulation of some key component(s) involved in caspase activation. PMID- 16242129 TI - The cyanobacterium Anabaena sp. PCC 7120 has two distinct beta-carotene ketolases: CrtO for echinenone and CrtW for ketomyxol synthesis. AB - Two beta-carotene ketolases, CrtW and CrtO, are widely distributed in bacteria, although they show no significant sequence homology with each other. The cyanobacterium Anabaena sp. PCC 7120 was found to have two homologous genes. In the crtW deleted mutant, myxol 2'-fucoside was present, but ketomyxol 2'-fucoside was absent. In the crtO deleted mutant, beta-carotene was accumulated, and the amount of echinenone was decreased. Therefore, CrtW catalyzed myxol 2'-fucoside to ketomyxol 2'-fucoside, and CrtO catalyzed beta-carotene to echinenone. This cyanobacterium was the first species found to have both enzymes, which functioned in two distinct biosynthetic pathways. PMID- 16242130 TI - Assembly of major histocompatibility complex class II subunits with invariant chain. AB - The highly polymorphic major histocompatibility complex class II (MHCII) polypeptides assemble in the ER with the assistance of invariant chain (Ii) chaperone. Ii binds to the peptide-binding pocket of MHCII heterodimers. We explored the mechanism how MHCII subunits attach to Ii. Expression with single alpha or beta subunits from three human HLA and two mouse H2 class II isotypes revealed that Ii co-isolates predominantly with the alpha polypeptide. Co isolation with alpha chain requires the groove binding Ii-segment and depends on M91 of Ii. Immunoprecipitation of Ii from pulse chase labeled cells showed sequential assembly of alpha and beta chains. PMID- 16242131 TI - Non-cell autonomous RNA silencing. AB - In plants and in some animals, the effects of post-transcriptional RNA silencing can extend beyond its sites of initiation, owing to the movement of signal molecules. Although the mechanisms and channels involved are different, plant and animal silencing signals must have RNA components that account for the nucleotide sequence-specificity of their effects. Studies carried out in plants and Caenorhabditis elegans have revealed that non-cell autonomous silencing is operated through specialized, remarkably sophisticated pathways and serves important biological functions, including antiviral immunity and, perhaps, developmental patterning. Recent intriguing observations suggest that systemic RNA silencing pathways may also exist in higher vertebrates. PMID- 16242132 TI - Cortisol responses and social buffering: a study throughout the life span. AB - The ability of specific adult females to moderate plasma cortisol responses throughout the life span was examined in male guinea pigs maintained in large mixed age/sex groups. At four critical life stages of social development (preweaning, periadolescent, sexually but not socially mature, and sexually and socially mature), the same male guinea pigs were exposed to the stressor of exposure to a novel environment for 4 h while either alone, with an unfamiliar adult female, or with a favored adult female, as based on objective criteria from behavioral observation at that life stage. In preweaning males (9-19 days of age), the favored female (biological mother), but not an unfamiliar female, reduced the cortisol response in the novel environment. In periadolescents (49-61 days), an unfamiliar female, but not the favored female, buffered the cortisol response. At the sexually but not socially mature stage (114-126 days), the cortisol response to novelty was depressed in all conditions, and not affected by either female. At the sexually and socially mature stage (270-330 days), the favored female, but not the unfamiliar female, moderated cortisol levels. These results corroborate previous findings in infants and full adults, demonstrate marked age-specific changes in the ability of females to buffer hypothalamic pituitary-adrenal responses, and identify a heretofore undescribed period of cortisol response suppression in maturing male guinea pigs. The changing pattern of social buffering during the life span described here for the guinea pig might represent a more general pattern for males of other group-living mammals. PMID- 16242133 TI - Hypogastric artery ligation for intractable pelvic hemorrhage. AB - OBJECTIVE: To assess the outcomes of bilateral hypogastric (internal iliac) ligation performed to control intractable pelvic hemorrhage and avoid hysterectomy. METHODS: A review of indications and outcomes for 117 cases of bilateral hypogastric artery ligation over 15 years (1990-2004). RESULTS: Apart from a slight lesion to the hypogastric vein, no complications were observed. Hemorrhage was effectively controlled in all 37 obstetric cases. In 13 of these cases, the uterus was preserved even when there was cervical pregnancy, placenta previa, placental abruption, uterine atony, and uterine rupture, and 4 women were delivered of mature infants. Hemorrhage was effectively controlled in 41 of 80 gynecologic cases. Prophylactic reduction of pelvic blood flow was the indication for the procedure in 39 cases, 5 of whom involving Jehovah's Witnesses adverse to blood transfusion. The uterus was preserved in only a few of the 41 controlled cases, but one woman (so far) was delivered of a mature infant. CONCLUSION: Hypogastric artery ligation was found to be indicated if (1) life-threatening pelvic hemorrhage could not be controlled by conservative methods; (2) prophylactic reduction of pelvic blood flow was needed to prevent anticipated hemorrhage; and (3) preservation of reproductive function was desired. The procedure was found to be safe and usually effective and should be taught during obstetric and gynecologic training. PMID- 16242134 TI - Effect of coffee intake on blood flow and maternal stress during the third trimester of pregnancy. AB - OBJECTIVE: To investigate the effects of maternal ingestion of an ordinary dose of coffee on maternal stress and placental and fetal blood circulation during the third trimester of pregnancy. METHODS: We performed a Doppler blood flow analysis for 10 women in the third trimester of pregnancy before and after they drank a cup of coffee. Salivary samples were collected from the 10 pregnant women and 14 nonpregnant controls just before coffee intake and 30 min later. Salivary cortisol levels and chromogranin A titers were determined. RESULTS: Coffee intake had no effect on maternal or fetal blood flow. Among the pregnant women, Salivary cortisol levels were significantly reduced after coffee intake but salivary chromogranin A concentration was not significantly different before and after coffee intake. CONCLUSION: The reduced salivary cortisol levels suggest that coffee intake decreases maternal stress during pregnancy. PMID- 16242135 TI - Comprehensive two-dimensional gas chromatography of polybrominated diphenyl ethers. AB - Comprehensive two-dimensional gas chromatography with micro electron-capture detection (GCxGC-muECD) was evaluated for the separation of 125 polybrominated diphenyl ethers (PBDEs). From among the six column combinations that were evaluated, DB-1x007-65HT was found to be the most suitable because of: (i) the highest number of BDE congeners separated; (ii) the least decomposition of higher brominated congeners; and (iii) the most suitable maximum operating temperature. The separation of the 125 BDE congeners from five hydroxy- and two methoxy-BDEs and nine other brominated flame retardants (polybrominated biphenyls, tetrabromobisphenol-A, methyl-tetrabromobisphenol-A and hexabromocyclododecane) was also studied. Fluorinated BDEs were found to be valuable internal standards for the determination of BDEs because of their very similar physico-chemical properties and excellent separation from the parent BDEs, mainly in the second dimension. GCxGC-time-of-flight MS and GCxGC-muECD were shown to be useful tools to identify decomposition products of nona- and deca-substituted BDEs, which are formed during the GC run. Three nona-BDEs were shown to be the major decomposition products of BDE 209. PMID- 16242136 TI - Facilitated column selection in pharmaceutical analyses using a simple column classification system. AB - In this paper, the performance of a previously developed classification system applied to pharmaceutical chromatographic analyses, is investigated. The separation of seven different drug substances from their respective impurities was studied. The chromatographic procedure for acetylsalicylic acid, clindamycin hydrochloride, buflomedil hydrochloride, chloramphenicol sodium succinate, nimesulide and phenoxymethylpenicillin was performed according to the corresponding European Pharmacopoeia (Ph. Eur.) monograph. The separation of dihydrostreptomycin sulphate was performed according to the literature. It is shown that the column ranking system is a helpful tool in the selection of a suitable column in these analyses. PMID- 16242137 TI - Study on interaction between poly(amidoamine) dendrimer and CdSe nanocrystal in chloroform. AB - The binding of different categories of molecules to quantum dot has been studied for many years through different spectroscopic techniques to elucidate details of binding mechanism. In this work we present the results of the study of the interactions between CdSe and poly(amidoamine) dendrimer monitored by photoluminescence spectroscopy of CdSe in chloroform. Dendrimers with different terminal groups and different generations were used to bind with CdSe nanocrystal of different size. Significant differences in the values of binding constant Kb(n) and K(SV) were found in these experiments. The binding constant for poly(amidoamine) dendrimer of generation 4.0 is higher as compared to generation 3.5. The interaction of CdSe with poly(amidoamine) dendrimer shows an increase of binding constants with increasing dendrimer generation from 2.0 to 4.0, as well as with decreasing CdSe diameter. From HRTEM and FTIR analysis, we suggest that dendrimer/CdSe interactions are primarily hydrogen-bonding. PMID- 16242138 TI - Transient electroosmotic flow induced by AC electric field in micro-channel with patchwise surface heterogeneities. AB - This paper investigates two-dimensional, time-dependent electroosmotic flow driven by an AC electric field via patchwise surface heterogeneities distributed along the micro-channel walls. The time-dependent flow fields through the micro channel are simulated for various patchwise heterogeneous surface patterns using the backwards-Euler time stepping numerical method. Different heterogeneous surface patterns are found to create significantly different electrokinetic transport phenomena. The transient behavior characteristics of the generated electroosmotic flow are then discussed in terms of the influence of the patchwise surface heterogeneities, the direction of the applied AC electric field, and the velocity of the bulk flow. It is shown that the presence of oppositely charged surface heterogeneities on the micro-channel walls results in the formation of localized flow circulations within the bulk flow. These circulation regions grow and decay periodically in phase with the applied periodic AC electric field intensity. The location and rotational direction of the induced circulations are determined by the directions of the bulk flow velocity and the applied electric field. PMID- 16242139 TI - The influence of water on the intercalation of epoxy monomers in Na montmorillonite. AB - The intercalation of a typical UV-curable epoxy monomer (CE) in unmodified montmorillonite and the effect of hydration on the intercalation reaction are studied. Montmorillonite in the sodium form was submitted to a controlled hydration/dehydration cycle and the water content was checked by TGA/XRD analyses. The structure of the hydrated Na+-montmorillonite was determined from the values of the basal spacings and from the water content of the hydrated form: a coordination of four water molecules per Na ion was found, corresponding to a minimum of energy calculated by molecular dynamics simulation. When dispersing the clay in the CE monomer, the anhydrous Na+-montmorillonite did not show any intercalation; on the contrary the hydrated form showed an increase of the basal spacing. A possible mechanism to explain the intercalation of the CE monomers is proposed. PMID- 16242140 TI - Relating clay yield stress to colloidal parameters. AB - The ability to predict rheological behavior of clay dispersions would be useful in formulating systems rheologically modified by clays as well as designing equipment to handle clay based materials. Deriving predictive equations for these materials has been hindered due to the various mechanisms by which clay systems develop microstructure. This work seeks to observe the relationship between the yield stress, tauy, and the zeta potential, zeta, and determine the nature of the particle interactions for dispersions of laponite (a synthetic clay) and dispersions of a mixture of naturally occurring kaolinites and bentonites. It is found that the relationship of tauy to zeta2 for clay suspensions is opposite to that found for homogeneously charged spheroidal colloids. This result can be traced to the type of particle interactions occurring, which for the systems studied appear to be edge-to-face attractions. PMID- 16242141 TI - Rapid ventricular pacing induces delayed cardioprotection against myocardial stunning. AB - Tachycardia with rapid ventricular pacing induces delayed preconditioning against arrhythmias secondary to coronary artery occlusion (CAO) and reperfusion (CAR) but its effects on myocardial stunning remains unknown. Accordingly, we investigated whether delayed preconditioning with ventricular pacing develops against myocardial stunning and whether this phenomenon is triggered by reactive oxygen species. Eight chronically instrumented conscious dogs underwent three experimental sequences in a random order a week apart: (a) 10-min CAO (coronary occluder) followed by CAR, i.e. "Control" sequence; (b) pacing (right ventricular electrodes, 240 beats/min during 40 min) performed 24 h before the 10-min CAO, i.e. "PC" sequence; and (c) N-(2-mercaptopropionyl)-glycine (MPG, 100 mg/kg per h) administered concomitantly to pacing and 10-min CAO performed 24 h later, i.e. "PC+MPG" sequence. During "Control", left ventricular (LV) wall thickening (%, sonomicrometry) was dramatically reduced during CAO (-96 +/- 5% from 2.9 +/- 0.4 mm) and remained depressed during CAR demonstrating myocardial stunning. During "PC", LV wall thickening was not altered by pacing per se and was similarly decreased during CAO vs. "Control". However, during CAR, LV wall thickening was improved vs. "Control" (e.g. -24 +/- 5% and -8 +/- 4% from corresponding baseline for "PC" and "Control", respectively at 2 h-CAR; P<0.05), demonstrating delayed preconditioning. Administration of MPG during pacing (n=5) abolished the beneficial effects of pacing. Myocardial lactate extraction and transmural distribution of regional myocardial blood flow (fluorescent microspheres) were not modified, by pacing. In conclusion, tachycardia with rapid ventricular pacing induces delayed cardioprotection against myocardial stunning. The production of reactive oxygen species independently from ischemia appears to be a major trigger for this phenomenon. PMID- 16242142 TI - P2 receptors in human heart: upregulation of P2X6 in patients undergoing heart transplantation, interaction with TNFalpha and potential role in myocardial cell death. AB - ATP acts as a neurotransmitter via seven P2X receptor-channels for Na(+) and Ca(2+), and eight G-protein-coupled P2Y receptors. Despite evidence suggesting roles in human heart, the map of myocardial P2 receptors is incomplete, and their involvement in chronic heart failure (CHF) has never received adequate attention. In left myocardia from five to nine control and 5-12 CHF subjects undergoing heart transplantation, we analyzed the full repertoire of P2 receptors and of 10 "orphan" P2Y-like receptors. All known P2Y receptors (i.e. P2Y(1,2,4,6,11,12,13,14)) and two P2Y-like receptors (GPR91 and GPR17) were detected in all subjects. All known P2X(1-7) receptors were also detected; of these, only P2X(6) was upregulated in CHF, as confirmed by quantitative real time PCR. The potential significance of this change was studied in primary cardiac fibroblasts freshly isolated from young pigs. Exposure of cardiac fibroblasts to ATP or its hydrolysis-resistant-analog benzoylATP induced apoptosis. TNFalpha (a cytokine implicated in CHF progression) exacerbated cell death. Similar effects were induced by ATP and TNFalpha in a murine cardiomyocytic cell line. In cardiac fibroblasts, TNFalpha inhibited the downregulation of P2X(6) mRNA associated to prolonged agonist exposure, suggesting that, by preventing ATP-induced P2X(6) desensitization, TNFalpha may abolish a defense mechanism meant at avoiding Ca(2+) overload and, ultimately, Ca(2+)-dependent cell death. This may provide a basis for P2X(6) upregulation in CHF. In conclusion, we provide the first characterization of P2 receptors in the human heart and suggest that the interaction between TNFalpha and the upregulated P2X(6) receptor may represent a novel pathogenic mechanism in CHF. PMID- 16242143 TI - The spectrum of cardiovascular anomalies in CHF1/Hey2 deficient mice reveals roles in endocardial cushion, myocardial and vascular maturation. AB - CHF1/Hey2 null mice generated in different laboratories have discrepant cardiovascular phenotypes. To determine the effect of genetic background on phenotype, we backcrossed our knockout strain more than eight generations to the inbred strains BALB/c and C57BL/6. Knockout mice on these backgrounds showed disparate phenotypes. Mice on both backgrounds demonstrated ventricular septal defects (VSDs), tricuspid stenosis and mitral valve thickening, but at varying frequencies, suggesting a general defect in endocardial cushion remodeling. Additional defects seen exclusively on the C57BL/6 background included biventricular wall thinning and left ventricular enlargement, implying a more severe myocardial defect than previously observed. In addition, aortas and pulmonary arteries from these null mice had thinner walls. Intercrossing of the CHF1/Hey2 null mice on a C57BL/6 background with a C57BL/6 MLC2v-CHF1/Hey2 transgenic line overexpressing CHF1/Hey2 in the atrial and ventricular myocardium also rescued the VSD and myocardial phenotypes, but did not affect vascular wall thickness. Our results indicate that CHF1/Hey2 provides an important myocardial signal to the endocardial cushion for proper septation and valve formation and also plays an important role in maturation of the myocardium and vasculature. PMID- 16242144 TI - Generation of metabolic oscillations by mitoKATP and ATP synthase during simulated ischemia in ventricular myocytes. AB - Metabolic oscillations and the concomitant periodic activations of sarcolemmal ATP-sensitive K(+) channels (sarcK(ATP)) have recently been proposed as one mechanism underlying ischemia-related arrhythmia. In this study, we investigated the role of mitochondrial ATP-sensitive K(+) channels (mitoK(ATP)) and ATP synthase in the generation of metabolic oscillations during simulated ischemia from rat ventricular myocytes using patch-clamp technique and fluorescence microscopy. We have found that the combined application of creatine kinase (CK) inhibitor, 2,4-dinitrofluorobenzene, with cyanide, electron-transport-chain inhibitor causes oscillatory activations of sarcK(ATP). The oscillatory activations of sarcK(ATP) were accompanied by large periodic depolarizations in mitochondrial membrane potential (Psi(m)). 5-Hydroxydecanoate, an inhibitor of mitoK(ATP), halted the oscillations in Psi(m) at repolarized state, whereas oligomycin, an inhibitor of ATP synthase, halted them at depolarized state. In both conditions, oscillatory activations of sarcK(ATP) were abolished. Inhibitors of adenine nucleotide translocator and permeability transition pore had no effect on the oscillations in Psi(m) and sarcK(ATP). 4,4'-diisothiocyanatostilbene-2,2' disulfonate, an inhibitor of mitochondrial inner-membrane anion channel (IMAC), caused a full depolarization in Psi(m) and activation of sarcK(ATP), finally resulting in irreversible hypercontracture. Taken together, oscillations in Psi(m) can be explained by balance between depolarizing power of mitoK(ATP) and repolarizing power of the reverse activity of ATP synthase. ATP consumption by ATP synthase in reverse mode links periodic depolarizations in Psi(m) to oscillatory activation of sarcK(ATP). Considering that such oscillations were not induced by cyanide alone, CK system may act as an important buffer, inhibiting arrhythmia during ischemia. PMID- 16242145 TI - Antiapoptotic effect of calcitonin gene-related peptide on oxidative stress induced injury in H9c2 cardiomyocytes via the RAMP1/CRLR complex. AB - Calcitonin gene-related peptide (CGRP) plays an important role in the mediation of protective effects observed in situations such as ischemic preconditioning in rat hearts. In this study, we investigated in H9c2 rat cardiomyoblasts if the protective effect of CGRP could be linked to an inhibitory effect on the apoptotic pathway. We also determined the specificity of observed effects by treatment with adrenomedullin (ADM) in stress conditions generated by 100 microM hydrogen peroxide. Using MTT assays, we demonstrate that a pretreatment with CGRP decreases by half the loss of cell viability induced by H(2)O(2). CGRP inhibits phosphatidylserine externalization, caspase 3 activation and DNA fragmentation due to oxidative stress. Using RT-PCR, we observed an increase in Bcl-2 mRNA expression induced by CGRP treatment. Dot blotting experiments showed that, in stress conditions, Bcl-2 protein level decreases while Bax is increased. CGRP administration prior to stress prevents these effects. The three-receptor activity modifying protein (RAMP) isotypes were detected by RT-PCR in H9c2 cells and in left ventricle rat tissue, RAMP1 and RAMP3 being the most abundant in both cases. RAMP1 expression was upregulated by CGRP while RAMP3 mRNA level was decreased. Cell viability assessed by MTT indicates that, contrary to CGRP, pretreatment of stressed cells with ADM, a RAMP2 agonist, fails to protect them while treatment with CGRP(8-37) (a RAMP1 and 2 inhibitor) abolished CGRP protective effect. Taken together, these data suggest that CGRP has antiapoptotic properties through the RAMP1/CRLR complex. CGRP could be used to prevent apoptosis in an ischemia-reperfusion context. PMID- 16242146 TI - Proliferation of cardiomyocytes derived from human embryonic stem cells is mediated via the IGF/PI 3-kinase/Akt signaling pathway. AB - Cardiomyocytes from common experimental animals rapidly exit the cell cycle upon isolation, impeding studies of basic cell biology and applications such as myocardial repair. Here we examined proliferation of cardiomyocytes derived from human and mouse embryonic stem (ES) cells. While mouse ES cell-derived cardiomyocytes showed little proliferation, human cardiomyocytes were highly proliferative under serum-free conditions (15-25% BrdU+/sarcomeric actin+). The cells exhibited only a small serum dose-response, and proliferation gradually slowed with increasing differentiation of the cells. Neither cell density nor different matrix attachment factors affected cardiomyocyte proliferation. Blockade of phosphatidylinositol 3-kinase (PI 3-kinase) and Akt significantly reduced cardiomyocyte proliferation, whereas MEK inhibition had no effect. Antibody blocking of the insulin-like growth factor-1 (IGF-1) receptor significantly inhibited cardiomyocyte proliferation, while addition of IGF-1 or IGF-2 stimulated cardiomyocyte proliferation in a dose-dependent manner. Thus, cardiomyocytes derived from human ES cells proliferate extensively in vitro, and their proliferation appears to be mediated primarily via the PI 3-kinase/Akt signaling pathway, using the IGF-1 receptor as one upstream activator. This system should permit identification of regulatory pathways for human cardiomyocyte proliferation and may facilitate expansion of cardiomyocytes from human ES cells for therapeutic purposes. PMID- 16242147 TI - Tachycardia increases NADPH oxidase activity and RyR2 S-glutathionylation in ventricular muscle. AB - We have shown previously that electrically induced tachycardia effectively produces myocardial preconditioning. Among other effects, tachycardia increases calcium release rates in microsomal fractions enriched in sarcoplasmic reticulum (SR) isolated from dog cardiac ventricular muscle. Here, we report that preconditioning tachycardia increased twofold the NADPH oxidase activity of isolated SR-enriched microsomal fractions, measured as NADPH-dependent generation of superoxide anion and hydrogen peroxide. Tachycardia also augmented the association of rac1 and the NADPH oxidase cytosolic subunit p47(phox) to the microsomal fraction, without modifying the content of the membrane integral subunit gp91(phox). Microsomes from control animals displayed endogenous S glutathionylation of cardiac ryanodine receptors (RyR2); in microsomal fractions isolated after tachycardia RyR2 S-glutathionylation levels were 1.7-fold higher than in controls. Parallel in vitro experiments showed that NADPH produced a transient increase in calcium release rates and enhanced 1.6-fold RyR2 S glutathionylation in control microsomes but had marginal or no effects on microsomes isolated after tachycardia. Catalase plus superoxide dismutase, and the NADPH oxidase inhibitors apocynin and diphenyleneiodonium prevented the in vitro stimulation of calcium release rates and RyR2 S-glutathionylation induced by NADPH, suggesting NADPH oxidase involvement. Conversely, addition of reducing agents to vesicles incubated with NADPH markedly inhibited calcium release and prevented RyR2 S-glutathionylation. We propose that tachycardia stimulates NADPH oxidase activity, which by enhancing RyR2 redox modifications such as S glutathionylation, would contribute to sustain faster calcium release rates during conditions of increased cardiac activity. This response may be an important component of tachycardia-induced preconditioning. PMID- 16242148 TI - Gene expression profile associated with chronic atrial fibrillation and underlying valvular heart disease in man. AB - Atrial fibrillation (AF) is the most common sustained cardiac arrhythmia in humans. The pathophysiology of AF involves electrical, structural and contractile remodeling, which is associated with changes in cardiac gene expression. Previous studies of gene-expression changes in clinical AF have mostly been limited to a small number of candidate genes and have not all been well controlled for underlying heart disease. The present study assessed AF-related gene-expression changes in valve-disease patients with microarrays representing the cardiac transcriptome. Right atrial appendages from 11 patients with chronic AF and underlying valvular heart disease (AF-VHD) and seven patients in sinus rhythm with VHD (SR-VHD) were individually compared to an age-matched sinus-rhythm control group (SR-CTRL, 11 patients) using cardiac-specific microarray analysis. One-class statistical analysis was used to identify genes differentially expressed between SR-VHD and SR-CTRL patients. Two-class statistical analysis was used to identify genes differentially expressed between AF-VHD and SR-VHD patients. Out of 3863 analyzed genes, 832 genes were differentially expressed between SR-VHD and SR-CTRL patients, and 169 genes were differentially expressed between AF-VHD and SR-VHD patients. Striking AF-related changes included altered expression of nine genes pointing towards the development of fibrosis (e.g. upregulation of transforming growth factor beta1), and changes in eight genes potentially related to an increased risk of thromboembolic events (e.g. upregulation of alpha2 macroglobulin). Microarray results were confirmed by quantitative PCR. Our results suggest that AF produces a characteristic profile of gene-expression changes that may be related to the pathophysiology of the arrhythmia. PMID- 16242149 TI - Isoproterenol does not enhance Ca-dependent Na/Ca exchange current in intact rabbit ventricular myocytes. AB - Cardiac Na/Ca exchange (NCX, NCX1.1) is critical in cardiac myocyte Ca regulation, and its altered function contributes to inotropic state, systolic dysfunction in heart failure and arrhythmogenesis. Regulation of NCX is multifaceted, but protein kinase A (PKA) effects on NCX function are controversial. Here, we use three different and complementary approaches to compare NCX function +/-1 microM isoproterenol (ISO) in intact rabbit cardiac myocytes (in paired comparisons). First, in field-stimulated intact cells we inferred the cytosolic [Ca] ([Ca](i)) dependence of NCX function from the decay rate of caffeine-induced [Ca](i) transients. Second, we measured caffeine-induced [Ca](i) and inward I(NCX) simultaneously (perforated patch voltage clamp), to measure directly the [Ca](i) dependence of NCX rate. Third, using whole cell ruptured patch with [Ca](i) heavily buffered to 100 nM, [Na](i)=10 mM, and I(Ca), SR Ca release and Na/K pump all blocked, we recorded I(NCX) ramps at 37 degrees C. We find that NCX function is not altered by PKA activation under any of these three protocols, where intracellular conditions ranged from near-physiological to highly controlled. This does not rule out NCX modulation by PKA under all conditions, or in species other than rabbit. However, such effects are likely to be either minor (vs. other PKA actions on myocyte Ca handling) or indirect, such as secondary effects dependent on altered local [Ca](i) and [Na](i). PMID- 16242150 TI - P66shc regulates endothelial NO production and endothelium-dependent vasorelaxation: implications for age-associated vascular dysfunction. AB - The p66shc adaptor protein mediates age-associated oxidative stress. We examined the role of p66shc in endothelial nitric oxide synthase (eNOS) signaling. Overexpression of p66shc inhibited eNOS-dependent NO production. RNAi-mediated down-regulation of endogenous p66shc led to activation of the proto-oncogene ras, and Akt kinase, with a corresponding increase in phosphorylation of eNOS at S1177 (S1179 on bovine eNOS). In rat aortic rings, down-regulation of p66shc suppressed the vasoconstrictor response to phenyephrine that was abrogated by treatment with the NOS inhibitor l-NAME, and enhanced vasodilation induced by sub-maximal doses of acetylcholine. These findings highlight a pivotal role for p66shc in inhibiting endothelial NO production, and endothelium-dependent vasorelaxation, that may provide important mechanistic information about endothelial dysfunction seen with aging. PMID- 16242152 TI - pH-dependent conformational flexibility of the SARS-CoV main proteinase (M(pro)) dimer: molecular dynamics simulations and multiple X-ray structure analyses. AB - The SARS coronavirus main proteinase (M(pro)) is a key enzyme in the processing of the viral polyproteins and thus an attractive target for the discovery of drugs directed against SARS. The enzyme has been shown by X-ray crystallography to undergo significant pH-dependent conformational changes. Here, we assess the conformational flexibility of the M(pro) by analysis of multiple crystal structures (including two new crystal forms) and by molecular dynamics (MD) calculations. The MD simulations take into account the different protonation states of two histidine residues in the substrate-binding site and explain the pH activity profile of the enzyme. The low enzymatic activity of the M(pro) monomer and the need for dimerization are also discussed. PMID- 16242151 TI - Crystal structure of a bacterial class 2 histone deacetylase homologue. AB - Histone deacetylases (HDACs) are among the most promising targets in cancer therapy. However, structural information greatly enhancing the design of HDAC inhibitors as novel chemotherapeutics has not been available on class 2 HDACs so far. Here we present the structure of the bacterial FB188 HDAH (histone deacetylase-like amidohydrolase from Bordetella/Alcaligenes strain FB188) that reveals high sequential and functional homology to human class 2 HDACs. FB188 HDAH is capable to remove the acetyl moiety from acetylated histones. Several HDAC-specific inhibitors, which have been shown to inhibit tumor activity in both pre-clinical models and in clinical trials, also inhibit FB188 HDAH. We have determined the crystal structure of FB188 HDAH at a resolution of 1.6 angstroms in complex with the reaction product acetate, as well as in complex with the inhibitors suberoylanilide hydroxamic acid (SAHA) and cyclopentyle-propionyle hydroxamic acid (CypX) at a resolution of 1.57 angstroms and 1.75 angstroms, respectively. FB188 HDAH exhibits the canonical fold of class 1 HDACs and contains a catalytic zinc ion. The highest structural diversity compared to class 1 enzymes is found in loop regions especially in the area around the entrance of the active site, indicating significant differences among the acetylated proteins binding to class 1 and 2 HDACs, respectively. PMID- 16242153 TI - Internucleotide movements during formation of 16 S rRNA-rRNA photocrosslinks and their connection to the 30 S subunit conformational dynamics. AB - UV light-induced RNA photocrosslinks are formed at a limited number of specific sites in the Escherichia coli and in other eubacterial 16 S rRNAs. To determine if unusually favorable internucleotide geometries could explain the restricted crosslinking patterns, parameters describing the internucleotide geometries were calculated from the Thermus thermophilus 30 S subunit X-ray structure and compared to crosslinking frequencies. Significant structural adjustments between the nucleotide pairs usually are needed for crosslinking. Correlations between the crosslinking frequencies and the geometrical parameters indicate that nucleotide pairs closer to the orientation needed for photoreaction have higher crosslinking frequencies. These data are consistent with transient conformational changes during crosslink formation in which the arrangements needed for photochemical reaction are attained during the electronic excitation times. The average structural rearrangement for UVA-4-thiouridine (s4U)-induced crosslinking is larger than that for UVB or UVC-induced crosslinking; this is associated with the longer excitation time for s4U and is also consistent with transient conformational changes. The geometrical parameters do not completely predict the crosslinking frequencies, implicating other aspects of the tertiary structure or conformational flexibility in determining the frequencies and the locations of the crosslinking sites. The majority of the UVB/C and UVA-s4U-induced crosslinks are located in four regions in the 30 S subunit, within or at the ends of RNA helix 34, in the tRNA P-site, in the distal end of helix 28 and in the helix 19/helix 27 region. These regions are implicated in different aspects of tRNA accommodation, translocation and in the termination reaction. These results show that photocrosslinking is an indicator for sites where there is internucleotide conformational flexibility and these sites are largely restricted to parts of the 30 S subunit associated with ribosome function. PMID- 16242154 TI - Posttraumatic stress disorder: a state-of-the-science review. AB - This article reviews the state-of-the-art research in posttraumatic stress disorder (PTSD) from several perspectives: (1) Sex differences: PTSD is more frequent among women, who tend to have different types of precipitating traumas and higher rates of comorbid panic disorder and agoraphobia than do men. (2) Risk and resilience: The presence of Group C symptoms after exposure to a disaster or act of terrorism may predict the development of PTSD as well as comorbid diagnoses. (3) Impact of trauma in early life: Persistent increases in CRF concentration are associated with early life trauma and PTSD, and may be reversed with paroxetine treatment. (4) Imaging studies: Intriguing findings in treated and untreated depressed patients may serve as a paradigm of failed brain adaptation to chronic emotional stress and anxiety disorders. (5) Neural circuits and memory: Hippocampal volume appears to be selectively decreased and hippocampal function impaired among PTSD patients. (6) Cognitive behavioral approaches: Prolonged exposure therapy, a readily disseminated treatment modality, is effective in modifying the negative cognitions that are frequent among PTSD patients. In the future, it would be useful to assess the validity of the PTSD construct, elucidate genetic and experiential contributing factors (and their complex interrelationships), clarify the mechanisms of action for different treatments used in PTSD, discover ways to predict which treatments (or treatment combinations) will be successful for a given individual, develop an operational definition of remission in PTSD, and explore ways to disseminate effective evidence-based treatments for this condition. PMID- 16242155 TI - Falls among older adults--risk factors and prevention strategies. PMID- 16242156 TI - Low dose (-)deprenyl is cytoprotective: it maintains mitochondrial membrane potential and eliminates oxygen radicals. AB - Hypoxia leads to a collapse in mitochondrial transmembrane potential (Deltapsi(M)), a fall in the ATP/ADP ratio, and finally cell death. Since ( )deprenyl directly modulates Deltapsi(M) and production of reactive oxygen species (ROS) by altering the respiratory function of mitochondria, we were interested in the dose-response relations of these effects. The changes in JC-1 red/green signal ratios {mitochondrial transmembrane potential}, and the changes in the cerium staining (intracellular ROS) in hypoxic and normoxic PC12 cell cultures were measured following 1 h of Argon hypoxia and 24 h of re-oxygenation in the absence and in the presence of various concentrations of (-)deprenyl. Deltapsi(M) shifted to lower values following hypoxia/re-oxygenation and all cells had decreased and uniform Deltapsi(M) levels. The amount of ROS increased. Following 24 h of treatment with various concentrations of (-)deprenyl during the re-oxygenation period, survival increased, the Deltapsi(M) shift caused by oxygen deprivation was reversed and the peroxy radical levels decreased except for at 10(-3) M. PMID- 16242157 TI - Natural cannabinoids: templates for drug discovery. AB - Recent studies have elucidated the biosynthetic pathway of cannabinoids and have highlighted the preference for a C-3 n-pentyl side chain in the most prominently represented cannabinoids from Cannabis sativa and their medicinally important decarboxylation products. The corresponding C-3 n-propyl side chain containing cannabinoids are also found, although in lesser quantities. Structure-activity relationship (SAR) studies performed on Delta(9)-tetrahydrocannabinol (Delta(9) THC), the key psychoactive ingredient of Cannabis, and its synthetic analogues have identified the C-3 side chain as the key pharmacophore for ligand affinity and selectivity for the known cannabinoid receptors and for pharmacological potency. Interestingly, the terminal n-pentyl saturated hydrocarbon side chain of endocannabinoids also plays a corresponding crucial role in conferring similar properties. This review briefly summarizes the biosynthesis of cannabinoids and endocannabinoids and focuses on their side chain SAR. PMID- 16242158 TI - Genistein abrogates pre-hemolytic and oxidative stress damage induced by 2,2' Azobis (Amidinopropane). AB - The pre-hemolytic mechanism induced by free radicals initiated from water-soluble 2,2'-azobis (2-amidinopropane) hydrochloride (AAPH) and its reversal by genistein was investigated in human erythrocytes. The time course of K+ efflux compared to the occurrence of hemolysis suggests that AAPH-induced hemolysis occurs indirectly via pore formation and band 3 oxidation as expected. However, genistein inhibited hemolysis, LDH release and membrane protein oxidation but not K+ efflux. This indicated that erythrocyte protein oxidation possibly in the hydrophobic core plays a significant role in the membrane pre-hemolytic damage. Chemiluminescence (CL) analysis carried out in non-lysed erythrocytes treated with AAPH showed a dramatic increase in CL indicating both reduced levels of antioxidants and increased membrane lipid peroxide. The V0 value was also increased up to 6 times, denoting a high degree of membrane peroxidation very early in erythrocyte membrane damage. The whole process was inhibited by genistein in a dose-dependent manner. These results indicate that the genistein inhibited both hemolysis and pre-hemolytic damage and also hindered membrane lipid peroxide formation and protein oxidation. In addition, it is suggested that pre-hemolytic damage is mediated mainly by the oxidation of both phospholipid and protein located in the deeper hydrophobic region of the membrane. PMID- 16242159 TI - Historical and ecological analysis of coral communities in Castle Harbour (Bermuda) after more than a century of environmental perturbation. AB - The coral reefs in Bermuda's Castle Harbour basin have been subjected to varying anthropogenic stressors for over 100 years. These include restriction of water flow through the construction of a causeway in the late 19th century and an extensive dredging and land reclamation operation during World War II. In the 1970s, disposal of bulk waste commenced at a foreshore reclamation site in Castle Harbour. Since 1996 the waste stream has included blocks of cement-stabilized municipal incinerator ash. This study provides a historical and quantitative ecological review of the Castle Habour reef ecosystem as a case study, assessing the responses of the reef to more than a century of anthropogenic disturbance. Measures of the coral community, flow rates, turbidity and sedimentary regimes suggest the present structure of the coral community largely reflects the impacts of the historic dredge and fill operations prior to the establishment of the foreshore dump site. Recent increases in the abundance of some sediment tolerant, massive reef-building coral species (Diploria strigosa and Montastraea cavernosa) suggest adaptation to chronic sediment stress. PMID- 16242160 TI - Heavy metal concentrations in Littorina saxatilis and Enteromorpha intestinalis from Manx Estuaries. PMID- 16242161 TI - SMC1 inhibition results in FRA3B expression but has no effect on its delayed replication. AB - Cellular processes involved in fragile site expression have been investigated by studying the effect on the replication pattern of the commonest fragile site FRA3B of RNA interference (RNAi)-mediated sister maintenance chromosome 1 (SMC1) inhibition in normal human fibroblasts. Replication timing of FRA3B in G2 was studied by bromodeoxyuridine (BrdU) labeling for the final 2h of cell culture whereas in the S phase was investigated by a fluorescence in situ hybridization (FISH)-based approach through the analysis of clones spanning the FRA3B region. Results showed that FRA3B is normally late replicated even though it is not expressed in untreated cells. On the other hand, SMC1 inhibition leads to FRA3B expression even if the percent of late replicated cells is comparable to control cells. These results obtained by analysing the commonest fragile site suggest that SMC1 plays a role in protecting late replicating regions from stresses occurring in the final steps of genome replication and that delayed replication is necessary but not sufficient for inducing fragile site expression. PMID- 16242162 TI - Trafficking of presynaptic AMPA receptors mediating neurotransmitter release: neuronal selectivity and relationships with sensitivity to cyclothiazide. AB - Postsynaptic glutamate AMPA receptors (AMPARs) can recycle between plasma membrane and intracellular pools. In contrast, trafficking of presynaptic AMPARs has not been investigated. AMPAR surface expression involves interactions between the GluR2 carboxy tail and various proteins including glutamate receptor interacting protein (GRIP), AMPA receptor-binding protein (ABP), protein interacting with C kinase 1 (PICK1), N-ethyl-maleimide-sensitive fusion protein (NSF). Here, peptides known to selectively block the above interactions were entrapped into synaptosomes to study the effects on the AMPA-evoked release of [3H]noradrenaline ([3H]NA) and [3H]acetylcholine ([3H]ACh) from rat hippocampal and cortical synaptosomes, respectively. Internalization of pep2-SVKI to prevent GluR2-GRIP/ABP/PICK1 interactions potentiated the AMPA-evoked release of [3H]NA but left unmodified that of [3H]ACh. Similar potentiation was caused by pep2 AVKI, the blocker of GluR2-PICK1 interaction. Conversely, a decrease in the AMPA evoked release of [3H]NA, but not of [3H]ACh, was caused by pep2m, a selective blocker of the GluR2-NSF interaction. In the presence of pep2-SVKI the presynaptic AMPARs on noradrenergic terminals lost sensitivity to cyclothiazide. AMPARs releasing [3H]ACh, but not those releasing [3H]NA, were sensitive to spermine, suggesting that they are GluR2-lacking AMPARs. To conclude: (i) release regulating presynaptic AMPARs constitutively cycle in isolated nerve terminals; (ii) the process exhibits neuronal selectivity; (iii) AMPAR trafficking and desensitization may be interrelated. PMID- 16242163 TI - In vitro and ex vivo distribution of [3H]harmane, an endogenous beta-carboline, in rat brain. AB - The endogenous beta-carboline, harmane, has been shown to bind to monoamine oxidase A (MAO-A) and a separate, high affinity, non-MAO site. Research in our laboratory has shown that harmane is an active component of clonidine-displacing substance (CDS), the proposed endogenous ligand for imidazoline binding sites (IBS). In the present study we have investigated the distribution of [3H]harmane in rat brain, and related the binding profile to the distribution of the MAO-A selective ligand [3H]Ro41-1049 and the I2BS ligand [3H]2-BFI. The in vivo distribution of [3H]harmane following intravenous administration was also investigated. Receptor autoradiography revealed a highly significant correlation for the distribution of [3H]harmane and [3H]Ro41-1049, and a significant correlation for [3H]harmane and the I2BS ligand [3H]2-BFI. The in vivo distribution of [3H]harmane suggests that the ligand accumulates in the adrenal gland and throughout the brain with the primary route of excretion occurring via the duodenum. In conclusion, these studies have shown that [3H]harmane labels a population of binding sites that reflect the distribution of MAO-A. Further evidence for a non-MAO, IBS [3H]harmane population has not been shown but the high level of expression of the MAO-A site is likely to have masked the much smaller population of I2BS. PMID- 16242164 TI - Selective inhibition of inducible nitric oxide synthase reduces neurological deficit but not cerebral edema following traumatic brain injury. AB - The role of inducible nitric oxide synthase (iNOS) in cerebral edema and neurological deficit following traumatic brain injury (TBI) is not yet clear-cut. Therefore, the aim of this study was to investigate the effect of three different iNOS inhibitors on cerebral edema and functional outcome after TBI. First, the time courses of blood--brain barrier (BBB) breakdown, cerebral edema, and neurological deficit were studied in a rat model of fluid percussion-induced TBI. The permeability of BBB to Evans blue was increased from 1 h to 24 h after TBI. Consistently, a significant increase in brain water content (BWC) was observed at 6 and 24 h post-TBI. A deficit in sensorimotor neurological functions was also observed from 6 h to 7 days with a maximum 24 h after TBI. Second, a single dose of aminoguanidine (AG; 100 mg/kg, i.p.), L-N-iminoethyl-lysine (L-NIL; 20 mg/kg, i.p.), or N-[3-(aminomethyl)benzyl]acetamide (1400W; 20 mg/kg, s.c.) was administered at 6 h post-TBI. Treatment with AG reduced by 71% the increase in BWC evaluated at 24 h, while L-NIL and 1400W had no effect. In contrast, the three iNOS inhibitors reduced the neurological deficit from 30% to 40%. Third, 1400W (20 mg/kg, s.c.) was administered at 5 min, 8 and 16 h post-TBI. Although this treatment paradigm had no effect on cerebral edema evaluated at 24 h, it significantly reduced the neurological deficit and iNOS activity. In conclusion, iNOS contributes to post-TBI neurological deficit but not to cerebral edema. The beneficial effect of iNOS inhibitors is not due to their anti-edematous effect, and the reduction of cerebral edema by AG is unlikely related to iNOS inhibition. The 6 h therapeutic window of iNOS inhibitors could allow their use in the treatment of functional deficit at the acute phase of TBI. PMID- 16242165 TI - Decreased accumulation of glutelin types in rice grains constitutively expressing a sunflower seed albumin gene. AB - Previous studies have shown differential accumulation of sulfur rich glutelins and sulfur poor prolamins in transgenic rice seeds expressing a sunflower seed albumin gene [Hagan, N.D., Upadhyaya, N., Tabe, L.M., Higgins, T.J., 2003. The redistribution of protein sulfur in transgenic rice expressing a gene for a foreign, sulfur-rich protein. Plant J 34, 1-11]. Here, we show, by two dimensional electrophoresis, differential accumulation of three classes of glutelin proteins - type I, II and III - and a globulin, not previously resolved, in transgenic seeds grown under low and high sulfur nutrition. Several glutelin polypeptides were resolved and four identified as a type I glutelin, two type II glutelins and a type III glutelin. Although sulfur nutrition did not affect the accumulation of sunflower seed albumin, the levels of all four identified glutelins and the globulin were lower in mature seeds derived from transgenic plants grown under sulfur-optimum or sulfur limited conditions compared to non transgenic rice seeds. The reduction of all four glutelin polypeptides and the globulin varied from 21% to 68%. The re-allocation of sulfur reserves from endogenous proteins to the sulfur sink in transgenic grain is suggestive of a transcriptional control of sulfur mobilization in plants. PMID- 16242166 TI - Do polyamines contribute to plant cell wall assembly by forming amide bonds with pectins? AB - Two new reducing glycoconjugates [N-D-galacturonoyl-putrescinamide (GalA-Put) and N,N'-di-D-galacturonoyl-putrescinamide (GalA-Put-GalA)] and homogalacturonan putrescine (GalAn-Put) conjugates were synthesised as model compounds representing possible amide (isopeptide) linkage points between a polyamine and either one or two pectic galacturonate residues. The amide bond(s) were stable to cold acid and alkali (2M TFA and 0.1M NaOH at 25 degrees C) but rapidly hydrolysed by these agents at 100 degrees C. The amide bond(s) were resistant to Driselase and to all proteinases tested, although Driselase digested GalAn-Put, releasing fragments such as GalA3-Put-GalA3. To trace the possible formation of GalA-polyamine amide bonds in vivo, we fed Arabidopsis and rose cell-cultures and chickpea internodes with [14C]Put. About 20% of the 14C taken up was released as 14CO2, indicating some catabolism. An additional approximately 73% of the 14C taken up (in Arabidopsis), or approximately 21% (in rose), became ethanol insoluble, superficially suggestive of polysaccharide-Put covalent bonding. However, much of the ethanol-inextractable 14C was subsequently extractable by acidified phenol or by cold 1M TFA. The small proportion of radioactive material that stayed insoluble in both phenol and TFA was hydrolysable by Driselase or hot 6M HCl, yielding 14C-oligopeptides and/or amino acids (including Asp, Glu, Gly, Ala and Val); no free 14C-polyamines were released by hot HCl. We conclude that if pectin-polyamine amide bonds are present, they are a very minor component of the cell walls of cultured rose and Arabidopsis cells and chickpea internodes. PMID- 16242167 TI - Characterization of a mitochondrial-targeting signal in the PB2 protein of influenza viruses. AB - Influenza virus RNA polymerase is a heterotrimeric complex consisting of PB1, PB2, and PA subunits. These polymerase subunits accumulate in the nucleus of infected cells. We report here that PB2, from both human and avian influenza viruses, could also localize to mitochondria in transfected cells. Importantly, cells infected with influenza A virus also displayed mitochondrial PB2. We show that an N-terminal motif composed of 120 amino acids is sufficient for localization of PB2 to mitochondria. In particular, leucine residues at positions 7 and 10 were essential for mitochondrial targeting. Recombinant influenza A/WSN/33 viruses expressing PB2 proteins with L7A and/or L10A mutations showed reduced viral titers, but unaffected levels of transcription, replication, and protein expression. The introduction of L7A and/or L10A mutations into recombinant viruses correlated with reduced mitochondrial membrane potential in infected cells, suggesting that mitochondrial localization of PB2 contributes to the preservation of mitochondrial function during influenza virus infection. PMID- 16242169 TI - Variability estimation of urban wastewater biodegradable fractions by respirometry. AB - This paper presents a methodology for assessing the variability of biodegradable chemical oxygen demand (COD) fractions in urban wastewaters. Thirteen raw wastewater samples from combined and separate sewers feeding the same plant were characterised, and two optimisation procedures were applied in order to evaluate the variability in biodegradable fractions and related kinetic parameters. Through an overall optimisation on all the samples, a unique kinetic parameter set was obtained with a three-substrate model including an adsorption stage. This method required powerful numerical treatment, but improved the identifiability problem compared to the usual sample-to-sample optimisation. The results showed that the fractionation of samples collected in the combined sewer was much more variable (standard deviation of 70% of the mean values) than the fractionation of the separate sewer samples, and the slowly biodegradable COD fraction was the most significant fraction (45% of the total COD on average). Because these samples were collected under various rain conditions, the standard deviations obtained here on the combined sewer biodegradable fractions could be used as a first estimation of the variability of this type of sewer system. PMID- 16242168 TI - The initial ocular following responses elicited by apparent-motion stimuli: reversal by inter-stimulus intervals. AB - Transient apparent-motion stimuli, consisting of single 1/4-wavelength steps applied to square-wave gratings lacking the fundamental ("missing fundamental stimulus") and to sinusoidal gratings, were used to elicit ocular following responses (OFRs) in humans. As previously reported [Sheliga, B. M., Chen, K. J., FitzGibbon, E. J., & Miles, F. A. (2005). Initial ocular following in humans: a response to first-order motion energy. Vision Research, in press], the earliest OFRs were strongly dependent on the motion of the major Fourier component, consistent with early spatio-temporal filtering prior to motion detection, as in the well-known energy model of motion analysis. Introducing inter-stimulus intervals (ISIs) of 10-200 ms, during which the screen was gray with the same mean luminance, reversed the initial direction of the OFR, the peak reversed responses (with ISIs of 20-40 ms) being substantially greater than the non reversed responses (with an ISI of 0 ms). When the mean luminance was reduced to scotopic levels, reversals now occurred only with ISIs > or=60 ms and the peak reversed responses (with ISIs of 60-100 ms) were substantially smaller than the non-reversed responses (with an ISI of 0 ms). These findings are consistent with the idea that initial OFRs are mediated by first-order motion-energy-sensing mechanisms that receive a visual input whose temporal impulse response function is strongly biphasic in photopic conditions and almost monophasic in scotopic conditions. PMID- 16242170 TI - Removal of selected pharmaceuticals, fragrances and endocrine disrupting compounds in a membrane bioreactor and conventional wastewater treatment plants. AB - Eight pharmaceuticals, two polycyclic musk fragrances and nine endocrine disrupting chemicals were analysed in several waste water treatment plants (WWTPs). A membrane bioreactor in pilot scale was operated at different solid retention times (SRTs) and the results obtained are compared to conventional activated sludge plants (CASP) operated at different SRTs. The SRT is an important design parameter and its impact on achievable treatment efficiencies was evaluated. Different behaviours were observed for the different investigated compounds. Some compounds as the antiepileptic drug carbamazepine were not removed in any of the sampled treatment facilities and effluent concentrations in the range of influent concentrations were measured. Other compounds as bisphenol A, the analgesic ibuprofen or the lipid regulator bezafibrate were nearly completely removed (removal rates >90%). The operation of WWTPs with SRTs suitable for nitrogen removal (SRT>10 days at 10 degrees C) also increases the removal potential regarding selected micropollutants. No differences in treatment efficiencies were detected between the two treatment techniques. As in conventional WWTP also the removal potential of MBRs depends on the SRT. Ultrafiltration membranes do not allow any additional detention of the investigated substances due to size exclusion. However, MBRs achieve a high SRT within a compact reactor. Nonylphenolpolyehtoxylates were removed in higher extend in very low-loaded conventional WWTPs, due to variations of redox conditions, necessary for the degradation of those compounds. PMID- 16242171 TI - Low-temperature anaerobic biological treatment of solvent-containing pharmaceutical wastewater. AB - Low-temperature or psychrophilic (<20 degrees C) anaerobic digestion (PAD) has recently been demonstrated as a cost-effective option for the treatment of a range of wastewater categories. The aim of this work was 2-fold: (1) to screen three anaerobic sludges, obtained from full-scale reactors, with respect to suitability for PAD of pharmaceutical-like, solvent-contaminated wastewater; (2) to assess the feasibility of PAD of this wastewater category. Toxicity thresholds of key trophic groups within three candidate biomass samples were assessed against solvents prevalent in pharmaceutical wastewaters (propanol, methanol and acetone). Specific methanogenic activity (SMA) assays indicated that the metabolic optimum of each candidate biomass was within the mesophilic range. One biomass sample exhibited higher SMA assays than the other candidate samples and was also the sample least methanogenically inhibited by the addition of solvents to batch cultures. This sludge was selected as the biomass of choice for laboratory-scale trials. Two identical expanded granular sludge bed (EGSB)-based anaerobic reactors were used for the treatment of solvent-contaminated wastewater at 15 degrees C, and at applied organic loading rates (OLRs) of 5-20 kg chemical oxygen demand (COD) m(-3)d(-1). COD removal efficiencies of 60-70% were achieved during the 450 day trial. In addition, SMA assays carried out at the conclusion of the trial indicated the development of a putatively psychrophilic hydrogenotrophic methanogenic community. PMID- 16242172 TI - Clearance rates of Sabella spallanzanii and Branchiomma luctuosum (Annelida: Polychaeta) on a pure culture of Vibrio alginolyticus. AB - The influence exerted by filter-feeding activity on bacterial density by two sabellid species from the Mediterranean Sea (Ionian Sea, Italy), Branchiomma luctuosum Grube and Sabella spallanzanii Gmelin (Annelida: Polychaeta) was investigated. Clearance rates and retention efficiencies were estimated utilizing the species Vibrio alginolyticus selected on account of previous field studies and its importance in fish culture pathogenicity. The Cmax was 43.2+/-2.63 L h( 1) g(-1) DW for B. luctuosum and 12.4+/-2.22 L h(-1) g(-1) DW for S. spallanzanii. The Retention efficiency was 98% corresponding to a removed bacterial biomass of 44.8+7.88 microgC L(-1) g(-1) DW for B. luctuosum and 70% corresponding to a bacterial biomass of 23.8+2.95 microgC L(-1) g(-1) DW for S. spallanzanii. Maximum retention was recorded after 20 min for the first species and after 30 min for the second one. Present laboratory experiments represent a contribution to the knowledge of the filtration activity of the two polychaetes, characterizing the filtration process on bacterioplankton. Both species resulted extremely efficient in removing V. alginolyticus from seawater in experimental tanks, thus confirming the previous data from the field studies and suggesting their employment as biofilters of microbially contaminated waters in intensive aquaculture. PMID- 16242173 TI - Studies on the phosphorus sorption capacity of substrates used in constructed wetland systems. AB - Langmuir sorption isotherm was used to screen various substrates for use in removing phosphorus (P) in constructed wetlands (CW). The nine tested substrates included four sands, two soils, bentonite, and two industrial by-products of furnace slag and fly ash. Results showed that the furnace slag had the highest P sorption capacity (8.89 g Pk g(-1)), followed was the fly ash (8.81 g P kg(-1)), and that of sand II was the lowest. Different kinds of sands also showed varying P sorption capacity (0.13-0.29 g P kg(-1)). P sorption capacity was influenced by both the physico-chemical characteristics of the substrates and the amount of organic matter (OM) added. Lifetime of sand II for P sorption estimated by Langmuir P sorption maximum was up to only 9 months in full-scale systems, while that of furnace slag could be used for up to 22 yr. Furnace slag has great potential as a CW substrate, due to its high P sorption capacity. The expected lifetime of constructed wetlands for P removal is strongly influenced by the choice of adsorbing substrate. PMID- 16242174 TI - Mucilage microcosms. AB - In the summers of 2000, 2001 and 2002, large amounts of sticky mucilaginous material aggregated to form masses of impressive dimensions over large areas of the Adriatic Sea, particularly in its northern part. Aggregates differing in size were sampled by SCUBA divers and submitted to chemical (nutrient and organic matter concentrations) and biological analysis (virus, bacteria and phytoplankton abundances and bacterial metabolism). Suspended and sinking mucilaginous aggregates were biota-rich environments where the abundance of planktonic organisms and the concentration of nutrients were orders of magnitude higher than in the surrounding seawater. The embedded phytoplankton was mostly composed of diatoms, but the dinoflagellate Gonyaulax fragilis, previously reported in association with the aggregates, was also present. A variety of processes occurred within the mucilaginous aggregates which resulted in the transformation of the organic matter composition and lability and contributed to a partial degradation of mucilage. For an efficient degradation of mucilage, several conditions are necessary: high bacterial abundance and activity and an efficient recycling of nutrients within the aggregates. Most of these conditions, appear to change depending on the type and age of the aggregate. During the first phase of aggregation (cobwebs and ribbons), bacterial activities addressed the degradation of organic matter, particularly that of the nitrogen fraction. The degradation products were rapidly taken up by bacteria, supporting an increase in their abundance and production. In aged mucilage (clouds), the degradation processes decreased and the bacterial metabolism suggested the presence of new organic labile compounds probably due to phytoplankton production. On the basis of our results, stringers, generally considered the first step of the aggregation process, seemed to be the result of a mechanical disruption of other types of aggregates. PMID- 16242175 TI - Relationships between heterotrophic bacteria and cyanobacteria in the northern Adriatic in relation to the mucilage phenomenon. AB - High variability of heterotrophic bacterial (HB; 0.1.10(9)-6.10(9) cells L(-1)), nanoflagellates (HNF; 0.02.10(6)-2.4.10(6) cells L(-1)) and cyanobacterial (CB; 10(6)-700.10(6) cells L(-1)) abundances were observed during approximately monthly measurements at six stations along the transect Po Delta-Rovinj from March 1999 to August 2002. Substantially higher values were observed during the stratification period (June-September) in the surface layer of the western stations that were under more direct influence of Po River discharges. Changes of the HB abundance were significantly correlated with temperature in the entire water column and chlorophyll a (Chla) concentration in the surface layer. The nutrients did not look to have directly influenced the HB growth, except orthophosphate in the eastern, more oligotrophic part of transect, where probably HB efficiently competed with phytoplankton for this nutrient. Temperature was also important for CB growth that, however, appeared to occur more intensively in waters with low nutrient concentrations. Probably, in these conditions CB can still develop, while larger autotrophs are strongly nutrient limited. In the upper water column of reduced salinity (35-37), the contributions of CB carbon biomass to the total picoplankton biomass (CB+HB) were mostly larger than 30% (up to 80%) in years with mucilage events (1991, 2000-2002) than in other years. For example, in 1999 only a few values were higher than 30% (up to 50%). In abundant presence of mucilaginous aggregates (e.g. in June 2000 and late June 2002) the chlorophyll a ascribed to CB accounted for much larger portions of total measured Chla. From these results it was concluded that cyanobacteria during mucilage events may play a substantially increased role both as primary producers and prey within the microbial loop of the northern Adriatic. PMID- 16242176 TI - Estimating population served by sewage treatment works from readily available GIS data. AB - Environmental risk assessment of household chemicals at the catchment scale requires an estimate of the load at individual Sewage Treatment Works (STWs). This can be achieved based upon population served and market consumption data. Although the population served is difficult to obtain, this paper shows that reasonable estimates can be made using readily available spatially referenced data. A new method is developed using STW data from the Exe and the Aire and Calder catchments and validated using 193 STWs within the Environment Agency's Anglian region. The estimated populations served were compared with available estimates of Population Equivalents (PEs). The population estimates were broadly similar to PEs for small works but agreement was lower for larger plants. The discrepancy for larger works is consistent with trade influent inclusion in the PE. The method is suitable for application to both rural areas and large urbanised areas, although the interpretation of corroborating data becomes increasingly difficult in very large urban areas serving more than one STW. PMID- 16242177 TI - Mercury in the environment and riverside population in the Madeira River Basin, Amazon, Brazil. AB - This work presents quality control results on the mercury concentrations in different environmental (river sediments, forest soils, river suspended matter and fish) and human samples from the lower Madeira River, Amazon sampled between 2001 and 2003, about 15-20 years after the nearly cessation of gold mining activities in the region, which reached its peak in the late 1980s. The study aimed to compare mercury concentrations in these environmental samples with those reported by other authors during the gold rush of the Madeira River Basin. Today, in the Madeira River the releases of mercury register a sudden reduction due the gold price fall in the international trade. However, about 100 t of Hg were released to the atmosphere and to aquatic systems in the region during the gold rush. The present survey shows that notwithstanding the reduction of Hg emissions to the Madeira River Basin from gold mining proper, concentrations in fish and humans are similar to those measured during the gold rush. Reduction in Hg concentrations is restricted to areas close to old point sources and only for abiotic compartments (air and sediments). Remobilization of Hg from bottom sediments plus re-emission from soils due to land use changes are probably responsible for keeping high Hg concentrations in biological samples. PMID- 16242178 TI - A methodology for the assessment of rehabilitation success of post mining landscapes--sediment and radionuclide transport at the former Nabarlek uranium mine, Northern Territory, Australia. AB - Protection of the environment post-mining is an important issue, especially where runoff and erosion can lead to undesirable material leaving post-mining landscapes and contaminating surrounding land and watercourses. Methods for assessment of the environmental impact and long-term behaviour of post-mining landforms based on scientific methodology are needed especially where field data are absent or poor. An appraisal of the former Nabarlek uranium mine was conducted to assess the site from a soil erosion perspective as part of an independent evaluation of overall rehabilitation success. Determination of the gross erosion occurring, sediment discharge to Cooper Creek and the resultant sediment associated radionuclide load in Cooper Creek were the primary objectives of the study. These objectives were achieved through the application of several models using parameter values collected from the site. The study found that the area containing the mill tailings repository is extremely stable and meets the guidelines established for long-term storage of uranium mill tailings. Most other areas on the site are stable; however there are some areas with a high sediment loss. Sediment concentration in Cooper Creek, which drains the site, was found to be within the Australian water quality guidelines for fresh water, however sediment concentrations in tributaries were found to exceed recommended levels. Radionuclide determinations on soil samples showed that the highest specific activities (Bq kg-1) were present on a small (0.44 ha) area with a relatively high erosion rate. This small area contributed the majority of the estimated flux to Cooper Creek of uranium-series radionuclides sorbed or structurally incorporated to eroded soil particles sourced from the mine site. This study provides a methodology for assessment of the erosional stability of such a landscape and consequent impact on water quality, using extensive field data and readily available and well known models and methodologies. PMID- 16242179 TI - Effect of doxazosin gastrointestinal therapeutic system on platelet degranulation and platelet-leukocyte microaggregate formation induced by physiologic shear stress in hypertension. AB - INTRODUCTION: In this prospective, ex vivo, single-blind study, the effect of doxazosin on platelet function was studied in patients with hypertension. MATERIALS AND METHODS: Platelet activation by shear stress was measured in whole blood samples of 22 hypertensive patients and 22 normotensive controls, using flow cytometry. Sheared samples were evaluated for CD62 expression, microaggregate formation, and Ca2+ mobilization. Results were collected at baseline and after 1 and 2 months of single-dose (4 mg/d) extended-release doxazosin gastrointestinal therapeutic system therapy. RESULTS: Doxazosin normalized blood pressure in hypertensive patients after 1 and 2 months of treatment. Hypertensive patients had a higher baseline percentage (mean+/-SD) of degranulated platelets (CD62+) than the normotensive control group (4.14+/-1.05 vs. 2.47+/-0.68, P<0.01). After 2 months of doxazosin gastrointestinal therapeutic system treatment, the percentage of CD62+ in the experimental group significantly decreased (P<0.05). At baseline, the number of platelet-leukocyte aggregates in vivo was greater in hypertensive patients (P<0.01); doxazosin did not normalize this measurement. Following shearing, platelet expression of CD62 increased significantly in the hypertensive group (P<0.001 vs. control). Shear stress-induced platelet activation and microaggregate formation were also greater in hypertensive patients. Intraplatelet-free calcium concentration was higher in hypertensive patients at baseline than in the normotensive group (P<0.001). At 2 months, doxazosin significantly reduced thrombin-stimulated Ca2+ mobilization in hypertensive patients (P<0.01 vs. baseline). CONCLUSIONS: Platelets from hypertensive patients are more readily activated by shear stress and demonstrate significant alterations in cytoplasmic-free calcium mobilization. Doxazosin treatment reduced blood pressure and normalized alterations in platelet function. PMID- 16242180 TI - Fast glucocorticoid actions on brain: back to the future. AB - Rapid, non-transcriptionally mediated, effects of glucocorticoids affect many behaviors as well as inhibition of function in the hypothalamo-pituitary-adrenal axis. In this short review, it is argued that the fast glucocorticoid actions which are mediated by membrane receptors are an ancient type of sterol/steroid mediated effect, and that these may be the primordial glucocorticoid receptors. Although the fast feedback actions of the glucocorticoids enjoyed study in the middle of the last century, new results and the availability of new techniques suggest that it is again time for a concerted effort to be made to understand the mechanism(s) of these rapid effects. PMID- 16242181 TI - Clinical aspects of sperm DNA fragmentation detection and male infertility. AB - Over the past 25 years, various methods have been developed to measure sperm DNA strand breaks in situ. Currently, there are four major tests of sperm DNA fragmentation, including the Comet, Tunel, sperm chromatin structure assay (SCSA) and the acridine orange test (AOT). The Comet assay is a light microscope technique where the sperm cells are mixed with melted agarose and then placed on a glass slide. The cells are lysed and then subjected to horizontal electrophoresis. The Tunel assay, another light microscope technique, transfers labeled nucleotide to the 3'OH group of a broken DNA strand with the use of terminal deoxynucleotidyl transferase. The fluorescence intensity of each scored sperm is determined as a "yes" or "no" for sperm on a light microscope slide or by channels of fluorescent intensity in a flow cytometer. The light microscope based AOT, uses the metachromatic properties of acridine orange to stain sperm cells. The SCSA treats sperm with low pH to denature DNA at the sites of DNA strand breaks, followed by acridine orange (AO) staining of green for native DNA and red for denatured DNA as measured by flow cytometry (FCM) as well as % sperm with high DNA stainability (HDS: immature sperm with intact DNA related to decreased fertilization rates). The SCSA method has defined a 27-30% DNA fragmentation index (DFI) as the point in which a man is placed into a statistical category of taking a longer time to in vivo pregnancy, intra uterine insemination (IUI) and more routine in vitro fertilization (IVF) cycles or no pregnancy. Current data suggest that intracytoplasmic sperm injection (ICSI) may help overcome the diminished pregnancy prognosis with high DFI over the other ART or natural methods. PMID- 16242182 TI - Effects of infauna harvesting on tidal flats of a coastal lagoon (Ria Formosa, Portugal): implications on phosphorus dynamics. AB - The systematic collection of benthic organisms in tidal flats of coastal lagoons should be taken into account for the management of these systems, once sediment disturbance affects biogeochemical processes by favouring pore water renewal during tidal inundation. The objective of the present work was to evaluate the effects of infauna harvesting on the phosphorus dynamics of muddy and sandy intertidal areas in the Ria Formosa. Sediment cores and overlying water were collected during August 2000 and February 2001 from reworked and undisturbed sediment before and after flooding. Results obtained showed that during the first minutes of flooding there was a marked decrease of phosphate in pore water of disturbed sediments. However, phosphate tidal fluxes from sandy sediment were clearly higher (17 nmol cm(-2) d(-1) in summer and 3 nmol cm(-2) d(-1) in winter) than in muddy sediment (0.4 nmol cm(-2) d(-1) in summer and -0.01 nmol cm(-2) d( 1) in winter). After muddy sediment disturbance concentrations of iron oxides increased quickly (from 5 to 16 micromol g(-1)) and phosphate was sorbed onto these iron oxides, resulting in a buffering of phosphate pore water concentrations at low values in the oxidized sediment zone. The estimated P output from muddy sediment decreased one to two orders of magnitude after sediment disturbance in contrast to sandy sediments in which the impact of infauna harvesting was minimal. Consequently, the P-cycle is influenced by the disruption of muddy habitats in tide-driven systems. Such information could be useful for the management of the lagoon. PMID- 16242183 TI - Structural and active site analysis of plasmepsins of Plasmodium falciparum: potential anti-malarial targets. AB - Comparative protein modeling, active site analysis and binding site specificity for the homologous series of plasmepsins (PM's), present in food vacuole of Plasmodium falciparum, are carried out. Four loops (L1, L2, L3 and L4), which show maximum structural deviations irrespective of type of inhibitor, have been identified. Comparison of the crystal structures of ligand complexes reveal that residues belonging to these loops have negligible coulomb and VDW interactions with the inhibitor but play major role in determining the openness of the binding cavity. The coulomb and VDW interactions between the PMII subsite pockets and inhibitors, which play a major role in determining the inhibition constants, are delineated. Besides small displacements, the catalytic residues D32 of PMII undergoes rotation around the Cgamma-Cbeta single bond to assist catalysis whereas side chain conformational deviations are not observed in D214 on plasmepsin activation. The mutant S79D of PMII (and the corresponding residues of PMI and PMIV) which helps in recognizing and cleaving substrates containing lysine at P1 position is surrounded by highly polar atmosphere stabilized by lysine. However, in PMIII significantly lower polar atmosphere around the mutant A78S/A78D is observed. Large buried side chain area of residues located at M15 and I289 of PMII (and corresponding residues of PMI and PMIV) corroborates well with increase in specificity constant for hydrophobic substrates. PMID- 16242184 TI - Cytotoxic and genotoxic effects of resin monomers in human salivary gland tissue and lymphocytes as assessed by the single cell microgel electrophoresis (Comet) assay. AB - Malignant tumors of the three major pairs and the numerous minor salivary glands in humans are rare, and little is known about their various etiologies. Considering the fact that resin monomers from dental restorative materials are released into the saliva and diffuse into the tooth pulp or gingiva, mucosa, and salivary glands, this may potentially contribute to tumorigenesis. Resin monomers may also be reabsorbed and reach the circulating blood as well. Whereas the cytotoxic potential of some components has been clearly documented, data on genotoxicity in human target cells require further investigation. In the present study, genotoxic and cytotoxic effects of three common methacrylates are investigated in human samples of salivary glands and peripheral lymphocytes. The Comet assay was used to quantify DNA single strand breaks, alkali labile and incomplete excision repair sites in salivary gland probes and lymphocytes of 10 volunteers. The xenobiotics investigated were triethyleneglycoldimethacrylate (TEGDMA), urethanedimethacrylate (UDMA), and 2-hydroxyethylmethacrylate (HEMA), with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) and dimethyl sulfoxide (DMSO) as controls. DNA migration was analyzed using the tail moment according to Olive (OTM). Cytotoxicity was monitored using trypan blue staining. With TEGDMA concentrations at 10(-5)m (10(-3)m), UDMA at 10(-7)m (10(-7)m), and HEMA at 10( 3)m (10(-5)m) significant enhancements of DNA migration were achieved in tissue cells (lymphocytes) as compared to the negative controls. At higher concentrations of up to 2.5x10(-2)m, induced DNA migration was expressed by OTM at 10.7 for TEGDMA in tissue cells (8.7 in lymphocytes), 10.5 for UDMA (6.4), and 9.7 for HEMA (6.1). The viability of the cell systems was not affected as concerns the threshold level for the assay of 75% viable cells except for the highest concentration tested for TEGDMA and UDMA in tissue cells. At higher concentration levels, all tested substances induced significant enhancement of DNA migration in the Comet assay as a possible sign for genotoxic effects in human salivary glands and lymphocytes. These data add to the results of prior studies in human peripheral lymphocytes and give evidence of a possible risk factor for tumor initiation in human salivary glands. PMID- 16242185 TI - Relationships between chemical composition, physical properties and transfection efficiency of polysaccharide-spermine conjugates. AB - Biodegradable water-soluble polysaccharide-spermine (SPM) polycation conjugates for nucleic acid delivery were synthesized by oxidizing polysaccharides using potassium periodate, followed by SPM conjugation. The polycations differ in their polysaccharide type, arabinogalactan (AG) or dextran (D), and/or in the IO(4)- /saccharide mole ratio used for polysaccharide oxidation (1:1, 1:3, or 1:5), resulting in either D(1:1)-SPM, AG(1:1)-SPM, D(1:3)-SPM, AG(1:3)-SPM, or AG(1:5) SPM. Chemical structure of the conjugates was characterized for total nitrogen and primary amino groups. Surface pH and electrical surface potential were determined by means of spectral changes of covalently attached 7-hydroxycoumarin (HC, a pH- and electrical surface potential-sensitive fluorophore). The binding and the electrostatic neutralization of the polycations by plasmid DNA, as well as the relationship between chemical structure, physical parameters, and transfection of NIH3T3 cells, were also studied. D(1:1)-SPM, the only polycation that showed efficient cell transfection in culture, was shown to have: (1) high SPM content (2000 nmol/mg); (2) high levels of cross-linked SPM (39-51%); (3) at DNA P-/NH3+ ratio of 2.0, a plateau in neutralization of cationic groups (+48 mV, as determined by HC-labeled D(1:1)-SPM titration with DNA), and a drop in zeta potential from +42 mV for the polymer alone to 0 mV for the polyplex, suggesting that some of the charges are hidden from the DNA; (4) pH(surface) value of 9.2, suggesting that at physiological bulk pH the polymer is only partially ionized, and therefore can act as a "proton sponge" in the endosome; and (5) high sensitivity to serum-rich growth medium. An oleyl derivative, N-oleyl-dextran spermine (ODS), was synthesized and demonstrated improved transfection efficiency in serum-rich medium. PMID- 16242186 TI - Is there a role for T-type Ca2+ channel in glioma cell proliferation? PMID- 16242188 TI - Modulation of sensory photophobia in essential blepharospasm with chromatic lenses. AB - PURPOSE: To determine the effect of photochromatic modulation with tinted lenses on the sensory symptoms of photophobia in blepharospasm patients. DESIGN: Nonrandomized case-control study. PARTICIPANTS: Thirty-four subjects (24 benign essential blepharospasm patients and 10 normal controls). METHODS: Subjects were seated in front of a calibrated light source. Beginning at 0%, the intensity of the light source was increased gradually until the patient reported symptomatic photophobia. The intensity of the light source was then measured with a light meter. This procedure was performed first with no chromatic lens and then with 7 different chromatic lenses, each blocking specific wavelengths of the visible spectrum. The subject was then asked which lens provided the greatest symptomatic improvement of photophobia. Statistical significance was calculated with analysis of variance and t test analysis. MAIN OUTCOME MEASURES: Objective measurement of light intensity tolerated and subjective assessment of photophobia were obtained for each chromatic lens tested. RESULTS: The light intensity tolerated by the normal subjects compared with the blepharospasm group was not statistically significant with no lens, but grew to 3.5 times that tolerated by the blepharospasm group as more of the higher wavelengths of the visible spectrum were blocked (P = 0.048). Lenses 4, 5, 6, and 7 allowed blepharospasm patients to tolerate a significantly higher intensity of light when compared with no lens (P = 0.04, P = 0.007, P = 0.03, and P = 0.01, respectively). Although the highest intensity of light tolerated was measured with lens 6, 71% of blepharospasm patients reported the greatest relief of photophobia with lens 7. CONCLUSIONS: Blepharospasm patients tolerate a lower intensity of light when compared with normal subjects; this differential in light tolerance becomes significantly more pronounced as the higher wavelengths of the visible spectrum are blocked. The symptoms of photophobia in blepharospasm patients can be reduced significantly with photochromatic modulation. Despite lens 6 allowing the patients to tolerate a higher intensity of light, the majority of patients preferred lens 7 for symptomatic relief of photophobia. These findings suggest that sensory photophobia may be related more to the wavelength than to the intensity of the light exposure. PMID- 16242187 TI - Calcium imaging of individual erythrocytes: problems and approaches. AB - Although in erythrocytes calcium is thought to be important in homeostasis, measurements of this ion concentration are generally seen as rather problematic because of the auto-fluorescence or absorption properties of the intracellular milieu. Here, we describe experiments to assess the usability of popular calcium indicators such as Fura-2, Indo-1 and Fluo-4. In our experiments, Fluo-4 turned out to be the preferable indicator because (i) its excitation and emission properties were least influenced by haemoglobin and (ii) it was the only dye for which excitation light did not lead to significant auto-fluorescence of the erythrocytes. From these results, we conclude that the use of indicators such as Fura-2 together with red blood cells has to be revisited critically. We thus utilized Fluo-4 in erythrocytes to demonstrate a robust but heterogeneous calcium increase in these cells upon stimulation by prostaglandin E(2) and lysophosphatidic acid. For the latter stimulus, we recorded emission spectra of individual erythrocytes to confirm largely unaltered Fluo-4 emission. Our results emphasize that in erythrocytes measurements of intracellular calcium are reliably possible with Fluo-4 and that other indicators, especially those requiring UV excitation, appear less favourable. PMID- 16242189 TI - Diffuse unilateral subacute neuroretinitis in Venezuela. AB - PURPOSE: To describe the clinical features and management of the largest reported series to date of patients with diffuse unilateral subacute neuroretinitis (DUSN). DESIGN: Observational case series. METHODS: The charts of all patients coded as having DUSN in a vitreoretinal clinic in Caracas, Venezuela between July 1979 and August 2000 were retrospectively reviewed. MAIN OUTCOME MEASURES: Demographic information, presenting visual acuity, clinical course, and treatment were evaluated. RESULTS: We identified 82 eyes of 78 patients with DUSN. The mean age at diagnosis was 16.7 years, significantly younger than the mean age of the Venezuelan population. Sixty-five (83.3%) patients were 20 years old or younger (P<0.001 when compared with the population of Venezuela). Thirty-three (42.3%) of the patients were female. The presenting visual acuity was 20/400 or worse in 69 eyes (84.1%). The subretinal nematode was identified in 33 eyes (40.2%). All nematodes were small, approximately 400 microm in length. CONCLUSIONS: In Venezuela, DUSN is seen primarily in young patients without a significant gender predilection. The vast majority of patients in this population presented with severe vision loss that was irreversible. PMID- 16242190 TI - High efficiency transfection of Plasmodium berghei facilitates novel selection procedures. AB - The use of transfection in the study of the biology of malaria parasites has been limited due to poor transfection efficiencies (frequency of 10(-6) to 10(-9)) and a paucity of selection markers. Here, a new method of transfection, using non viral Nucleofector technology, is described for the rodent parasite Plasmodium berghei. The transfection efficiency obtained (episomal and targeted integration into the genome) is in the range of 10(-2) to 10(-3). Such high transfection efficiency strongly reduces the time, number of laboratory animals and amount of materials required to generate transfected parasites. Moreover, it allows different experimental strategies for reverse genetics to be developed and we demonstrate direct selection of stably and non-reversibly transformed, fluorescent protein (FP)-expressing parasites using FACS. Since there is no need to use a drug-selectable marker, this method increases the (low) number of selectable markers available for transformation of P. berghei and can in principle be extended to utilise additional FP. Furthermore the FACS-selected, FP expressing parasites may serve as easily visualized reference lines that may still be genetically manipulated with the existing drug-selectable markers. The combination of enhanced transfection efficiency and a versatile rodent model provides a basis for the further development of novel tools for high throughput genome manipulation. PMID- 16242192 TI - Prevalence of anemia in chronic obstructive pulmonary disease: comparison to other chronic diseases. AB - Chronic obstructive pulmonary disease (COPD) is a multisystemic inflammatory disease characterized by pulmonary and extrapulmonary symptoms. The impaired lung function has long-term implications on metabolism and homeostasis of many organ systems such as the skeleton, heart, brain and skeletal muscle. The occurrence and prevalence of anemia in COPD has rarely been studied. Anemia is such a common and simple clinical finding that we may underestimate its physiological relevance in COPD. The aim of the study was to retrospectively investigate the prevalence of anemia in a large population of COPD patients and to compare it to patients with chronic heart failure, renal insufficiency, cancer and asthma. A population of 7337 patients that was treated in the University Hospital Charite, Berlin, Germany, from 1996 to 2003 was subsetted according to the ICD-9/10 code of the discharge diagnoses into the above-mentioned diagnoses groups. The overall prevalence of anemia in COPD patients was 23.1%. It was comparable to the prevalence of anemia we found in patients with chronic heart failure (23.3%). Patients with renal insufficiency and cancer presented the highest anemia frequencies. The high prevalence of anemia in hospitalised COPD patients that were treated mostly for exacerbations gives evidence that anemia is also a comorbidity in COPD and may contribute to exercise limitation and dyspnoea. PMID- 16242191 TI - Glycosylated inositol phospholipid from Entamoeba histolytica: identification and structural characterization. PMID- 16242193 TI - Chemically modified tetracyclines (CMT-3 and CMT-8) enable control of the pathologic remodellation of human aortic valve stenosis via MMP-9 and VEGF inhibition. AB - OBJECTIVE: Tetracycline derivatives affect many cellular functions relevant to chronic cardiovascular pathologies, including cell proliferation, migration and matrix remodelling. Accordingly, we sought to determine whether they may modulate the pathologic characteristics known to be significantly involved in human aortic valve stenosis, such as gelatinase production, apoptosis, expression of vascular endothelial growth factor (VEGF) and tumour necrosis factor-alpha (TNF-alpha). METHODS: The effects of tetracycline derivatives (tetracycline and CMTs-3, -5, 8) on MMP-2 and -9 and their endogenous tissue inhibitor (TIMP-1 and -2) production profiles in explanted human aortic valve pieces were examined by means of gelatine zymography and reverse zymography. Chemiluminescent ELISA was performed to assess VEGF and TNF-alpha concentrations in the medium, and in order to evaluate programmed cell death, in situ labelling of the 3'-ends of the DNA fragments generated by apoptosis-associated endonucleases was performed. RESULTS: CMT-3 and -8 lowered the MMP-9 and VEGF levels significantly in a drug-, dose-, and time-dependent manner. MMP-2 and TIMPs remained unchanged, emphasizing the specificity of CMTs to MMP-9 production on the one hand and restoring the beneficial equilibrium of MMP-9 and TIMPs on the other. Tetracycline was the only drug with a significant impact on net gelatinolytic activity, suggesting that the effect of tetracycline is more extensive concerning total MMP activity. CONCLUSIONS: Tetracycline derivatives may have therapeutic effects on the pathologic remodellation of advanced human aortic stenosis through the inhibition of MMP-9 and VEGF production. PMID- 16242195 TI - Percutaneous treatment of ventricular tachycardia, perimembranous ventricular septal defect and patent foramen ovale: a case report. PMID- 16242194 TI - Adults with Ebstein's anomaly--Cardiopulmonary exercise testing and BNP levels exercise capacity and BNP in adults with Ebstein's anomaly. AB - INTRODUCTION: Ebstein's anomaly is defined as the significant apical displacement of the part of the tricuspid valve causing significant tricuspid regurgitation and reduction of the functional right ventricle. The aim of the study was to evaluate exercise capacity with cardiopulmonary stress testing and to determine plasma BNP levels in adults with Ebstein's anomaly, and to establish their relation with echocardiogaphic grading of the lesion severity. MATERIALS AND METHODS: Study group consisted of 21 patients (16 males, aged 40.3+/-11.5 years). The control group: 19 healthy individuals (13 males, aged mean 39.9+/-9.3 years). On echocardiography the grade of the lesion severity was calculated (EGE) and used to define the following four groups: I < 0.5, II: 0.5-0.9, III: 1.0-1.49, IV > 1.5. The forced vital capacity (FVC), first second forced expiratory volume (FEV1), peak oxygen uptake (peak VO2), and VE/VCO2 slope were assessed with cardiopulmonary stress test and plasma BNP levels measured with radioimmunometric assay. RESULTS: In the studied group VO2 was lower than in control (21.9+/-5.4 vs. 33.6+/-8.3 mL/kg/min [p = 0.00001]), VE/VCO2 slope was higher in Ebstein's group (40.1+/-8.1, p = 0.00001). BNP levels were higher in the Ebstein group then in controls (35.9+/-25.0 vs. 17.2+/-9.9 pg/mL [p = 0.0002]) and did not differ significantly between EGE groups. PeakVO2 of 24.5+/-3.9 in patients from II EGE group were higher than in patients from EGE groups: III (17.2+/-5.2 p = 0.007) and IV (22.9+/-4.7 p = 0.05). CONCLUSIONS: Exercise capacity of adults with Ebstein's anomaly is significantly reduced and plasma BNP levels are higher compared to healthy individuals. Exercise capacity in patients with Ebstein's anomaly becomes gradually lower alongside the EGE severity; however, BNP levels do not correlate significantly with this parameter. PMID- 16242196 TI - Description and first results of a BVDV control scheme in Brittany (western France). AB - A collective zoo-sanitary control scheme for BVDV has been implemented in Brittany in successive steps since 1998. Prior evaluation of prevalence and dynamics of infection through bulk-milk ELISA showed that 40% of the dairy herds were likely to be free of BVDV but that, in the long term, no improvement was to be expected without control of new infections. The next step was a test-and-cull programme for PI animals in ELISA positive herds. Ten percentage of the herds were found to have at least one PI animal. The incidence decreased during this step. To lower costs of testing traded animals, a database was developed to collect all the available information to trace animals already known to be non PI. PMID- 16242197 TI - Ear-tag retention and identification methods for extensively managed water buffalo (Bubalus bubalis) in Trinidad. AB - Thirty-two young domestic water buffalo were studied to evaluate ear-tag retention during an epidemiologic field trial. Plastic ear-tags were placed in both ears before the start of the trial, which was implemented in an extensively managed domestic water buffalo herd of approximately 1000 animals in Trinidad from 1999-2001. The presence or absence of ear-tags was recorded at the times of animal handling. The rate of ear-tag loss was modeled using a parametric survival analysis assuming an exponential rate of loss. A gamma distribution was used to estimate the amount of time that each animal would be positively identified based only on the presence or absence of one or more ear-tags. The overall median ear tag retention was 272 days. The estimated rate of ear-tag loss was 0.0024 ear tags lost per day. The use of ear-tags alone might not be sufficient for long term identification of extensively managed animal populations. PMID- 16242198 TI - Three- to eight-month-old infants' catching under monocular and binocular vision. AB - We report a cross-sectional and a longitudinal experiment that examined developmental changes in the relative contribution of monocular and binocular variables in the guidance of interceptive arm movements. Three- to eight-month old infants were observed while presented with differently sized balls that approached frontally with a constant velocity under both monocular and binocular viewing conditions. Movement onset indicated that with age infants increasingly came to rely on binocular variables in controlling the timing of the interceptive arm movements. That is, from 7 to 8 months of age movement onset was independent from object size under binocular but not under monocular viewing. In contrast, binocular viewing enhanced the spatial accuracy of the interceptive arm movements at all ages. We concluded that attunement to binocular information is a key process in infants' gaining adaptive control of goal-directed arm movements. However, interceptive arm movements entail the formation of multiple on-line couplings between optic and movement variables, each of which appears to develop at its own pace. PMID- 16242199 TI - Assessment of distributions for fitting lag times of individual cells in bacterial populations. AB - To develop mathematical models describing lag times of individual bacterial cells (tau), experimental tau data were fitted to a variety of continuous distributions using BestFit. Six strains of Escherichia coli O157:H7 were used, and serial dilutions were made in Bioscreen multi-well plates to get single cells per well. Detection times (td) for individual wells were converted to tau using the maximum specific growth rate (mu) for each strain. All strains were subject to 25 trials, with up to 100 replicate wells per trial. Some strains had significantly longer t(d), and lower mu, but the tau values were not significantly different. Distributions were best fit in the order Pearson V>Pearson VI>Extreme Value>Lognormal>Lognormal2>Inverse Gaussian based on the Anderson-Darling statistic. The Lognormal distribution was selected because there was less variability in the parameter values, and parameters have specific biological meanings. Distributions could be fit to sample populations as low as six, with fittings and parameter values comparable to those obtained with larger samples (up to 89). Extreme Value, Pearson V, and Pearson VI distributions were more suitable for fitting tau values generated from a Lognormal distribution when the numbers of sample points were few, which suggested that there are similarities between the distributions. The results suggest that a Lognormal distribution can be used successfully to characterize tau. PMID- 16242200 TI - Glucose oxidase immobilization on a novel cellulose acetate polymethylmethacrylate membrane. AB - Glucose oxidase (GOD) was immobilized on cellulose acetate-polymethylmethacrylate (CA-PMMA) membrane. The immobilized GOD showed better performance as compared to the free enzyme in terms of thermal stability retaining 46% of the original activity at 70 degrees C where the original activity corresponded to that obtained at 20 degrees C. FT-IR and SEM were employed to study the membrane morphology and structure after treatment at 70 degrees C. The pH profile of the immobilized and the free enzyme was found to be similar. A 2.4-fold increase in Km value was observed after immobilization whereas Vmax value was lower for the immobilized GOD. Immobilized glucose oxidase showed improved operational stability by maintaining 33% of the initial activity after 35 cycles of repeated use and was found to retain 94% of activity after 1 month storage period. Improved resistance against urea denaturation was achieved and the immobilized glucose oxidase retained 50% of the activity without urea in the presence of 5M urea whereas free enzyme retained only 8% activity. PMID- 16242201 TI - Stress-related differential expression of multiple beta-carotene ketolase genes in the unicellular green alga Haematococcus pluvialis. AB - The unicellular green alga Haematococcus pluvialis is used as a biological production system for astaxanthin. It accumulates large amounts of this commercially interesting ketocarotenoid under a variety of environmental stresses. Here we report the identification and expression of three different beta-carotene ketolase genes (bkt) that are involved in the biosynthesis of astaxanthin in a single strain of the alga. Bkt1 and bkt2 proved to be the crtO and bkt previously isolated from two different strains of H. pluvialis. Bkt3 is a novel third gene, which shared 95% identical nucleotide sequence with bkt2. Nitrogen deficiency alone could not induce the alga cells to produce astaxanthin in 3 days even though it enhances the expression of the bkt genes to three times of that in normal growing cells within 16 h. High light irradiation (125 micromol m(-2)s(-1)) or 45 mM sodium acetate greatly increased the expression of bkt genes to 18 or 52 times of that in normal growing cells, resulting in an accumulation of substantial astaxanthin (about 6 mg g(-1) dry biomass) in 3 days. It is suggested that the existence of the multiple bkt genes and their strong up regulation by different stress conditions is one of the reasons that H. pluvialis accumulates large amounts of astaxanthin in an instant response to stress environments. PMID- 16242202 TI - Influence of initial pH on hydrogen production from cheese whey. AB - Batch experiments were conducted to investigate the effect of initial pH, between 5 and 10, on fermentative hydrogen production from crude cheese whey (87.5% (v/v) by Clostridium saccharoperbutylacetonicum). Hydrogen was produced over the range of pH studied. The hydrogen production rate and yield peaked at an initial pH 6 and then steadily decreased as the pH increased. The highest rate and yield were 28.3 mlh(-1) and 7.89 mmolg(-1) lactose, respectively. Sugar consumption was unaffected between pH 5 and 9 and remained at 97%. All final pHs were acidic and increased alongside the initial pH. There was no correlation between the initial pH and the fermentation time; the times were shorter (50-52 h) between pH 6 and 8, and longer (62-82 h) outside this range. A modified Gompertz equation adequately described fermentative hydrogen production from cheese whey. The respective maximum hydrogen production rate and hydrogen potential at an optimal pH of 6 were 47.07 mlh(-1) and 1,432 ml. Lag phase times were much longer at acidic pHs than at alkaline pHs. PMID- 16242204 TI - Chromosome evolution in eukaryotes: a multi-kingdom perspective. AB - In eukaryotes, chromosomal rearrangements, such as inversions, translocations and duplications, are common and range from part of a gene to hundreds of genes. Lineage-specific patterns are also seen: translocations are rare in dipteran flies, and angiosperm genomes seem prone to polyploidization. In most eukaryotes, there is a strong association between rearrangement breakpoints and repeat sequences. Current data suggest that some repeats promoted rearrangements via non allelic homologous recombination, for others the association might not be causal but reflects the instability of particular genomic regions. Rearrangement polymorphisms in eukaryotes are correlated with phenotypic differences, so are thought to confer varying fitness in different habitats. Some seem to be under positive selection because they either trap favorable allele combinations together or alter the expression of nearby genes. There is little evidence that chromosomal rearrangements cause speciation, but they probably intensify reproductive isolation between species that have formed by another route. PMID- 16242203 TI - Psammomys obesus (sand rat), a new animal model of non-alcoholic fatty liver disease. AB - BACKGROUND/AIMS: Non-alcoholic fatty liver disease (NAFLD) is an increasingly recognized condition that includes a spectrum of clinicopathologic conditions ranging from steatotosis to cirrhosis and liver failure. NAFLD is usually associated with features of the metabolic syndrome. No established therapies can be offered to patients with NAFLD. An appropriate animal model of NAFLD would be of help in understanding the mechanisms of the disease and in testing novel therapeutic modalities. Available animal models, such as ob/ob and db/db mice, are unsatisfactory since they show only partial resemblance to human NAFLD. Psammomys obesus (sand rat) is a well-established model of type-2 diabetes and obesity, which shares most metabolic parameters of the human metabolic syndrome. In the present study, we hypothesized that P. obesus will also show features of non-alcoholic fatty liver disease. METHODS: Experimental and control animals were fed normal rat chow or either chow to which fiber (30% wheat straw) was added for 6-18 weeks. Body weight and capillary glucose were measured regularly. At sacrifice blood samples, liver and epididymal fat were obtained. Histology of the liver was blindly determined by a pathologist. RESULTS: The experimental group showed increased body weight, liver and abdominal fat pad mass, raised plasma glucose, insulin and lipids. Also, alanine-aminotransferase (189+/-76 IU versus 86+/-26 IU; p<0.0001) was significantly higher in the experimental than the control group. Microscopic examination of liver tissue demonstrated marked macrovesicular fat infiltration in the experimental group while it was histologicaly normal in the control animals (liver fat score 1.7+/-1.0 and 0.2+/ 0.4; p<0.0001, respectively). CONCLUSIONS: Fed a calorie-rich diet P. obesus develops a syndrome, which shares metabolic, laboratory and histopathologic characteristics compatible with human NAFLD. PMID- 16242205 TI - Correction: high-level disinfection of cystoscopic equipment with ortho phthalaldehyde solution. PMID- 16242206 TI - Surveillance of nosocomial infections in paediatric recipients of bone marrow or peripheral blood stem cell transplantation during neutropenia, compared with adult recipients. AB - The aim of this prospective study was to analyse the incidence of nosocomial infections (NIs) during neutropenia after haematopoietic stem cell transplantation (HSCT) in paediatric and adult patients. Diagnosis of NI followed the modified surveillance protocol of the German National Reference Centre for Surveillance of NIs. During the 24-month study period, 38 and 39 transplantations were performed in paediatric and adult patients, respectively. Eighty percent and 92% of the paediatric and adult patients, respectively, underwent allogenic HSCT. In total, 1156 neutropenic days were documented. The incidence of NI during neutropenia in all recorded cases was 38.9 NI per 1000 days, without significant difference between the groups. No difference was found in the incidence rates of pneumonia, while there was a trend towards a higher incidence of bloodstream infection in adult patients. Significantly more episodes of fever of unknown origin were found in paediatric than adult patients. Pathogens were isolated in 19 of 45 cases of NI. Of the clinically documented infections, Gram-positive organisms were isolated in 79%, Gram-negative organisms were isolated in 16% and fungi were isolated in 5%. Surveillance of NI is an effective instrument to control the quality of health care. It should focus on device-associated infections or specific high-risk targets such as nosocomial pneumonia. PMID- 16242207 TI - Why investigate nosocomial diarrhoea? PMID- 16242208 TI - A new screening method for carbapenem-resistant Acinetobacter baumanii and Pseudomonas aeruginosa. PMID- 16242211 TI - Environmental sampling for MRSA: a note of caution. PMID- 16242209 TI - Bacterial insertion sequence IS256 as a potential molecular marker to discriminate invasive strains from commensal strains of Staphylococcus epidermidis. AB - The skin commensal Staphylococcus epidermidis has become one of the most important causative agents of nosocomial infections associated with medical devices. Differentiation between invasive S. epidermidis and its commensal counterpart is crucial for clinical decision making. The ica gene locus, which codes for production of polysaccharide intercellular adhesion (PIA), represents a frequently suggested molecular marker for infectivity. Our data demonstrated that production of PIA was not significantly increased among clinical strains, which may explain the controversial results obtained previously on the correlation of ica presence with origin from infection. Therefore, in this study, we attempted to identify novel genes discriminating between invasive and commensal strains based on the comparison of genome sequences. Our results indicated that the bacterial insertion sequence element IS256 occurred significantly more frequently in strains of clinical origin. Importantly, IS256 might thus constitute a molecular marker to discriminate invasive strains from commensal strains of S. epidermidis. PMID- 16242210 TI - Microcyn: a novel super-oxidized water with neutral pH and disinfectant activity. AB - A new super-oxidized water (SOW) product, Microcyn, was tested for in vitro antimicrobial and antiviral activities. The effectiveness of this neutral-pH SOW at killing Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Salmonella typhi and Candida albicans in pure culture was evaluated. One millilitre (approximately 10(8)colony-forming units/mL) of each micro-organism was subjected to 9 mL Microcyn or sterile water at room temperature for 30s. Under these conditions, a log(10) reduction factor of 8 in the level of all pathogens occurred in the treatment samples. In addition, results of tests with three batches of Microcyn exposed to Bacillus atrophaeus spores for 5 min demonstrated complete inactivation of the spores within 2-3 min (log(10) reduction factor >4). The effectiveness of Microcyn in reducing human immunodeficiency virus-1 (HIV-1) on hard surfaces (glass) was also evaluated in compliance with Environmental Protection Agency requirements for virucidal claims. After exposure of the tested surfaces to Microcyn for 5 min without agitation, there was a log(10) reduction factor >3 in the viral load as measured by both cytopathic effect and antigen p24 of HIV-1 production in MT-2 cultures. Microcyn activity against adenoviral vector type 5 was also analysed under simulated laboratory in-use conditions with viral suspensions. In order to increase the sensitivity of the test, the fluorescent light emitted by AdGFP infected cells was measured with the use of a flow cytometer. A log(10) reduction factor >3 in the viral load was achieved after a 5-min exposure to Microcyn under these strict conditions. These results show that Microcyn exerts a wide antimicrobial spectrum with major advantages over acidic SOWs, including neutral pH, lower free active chlorine (51-85 ppm) and long shelf life (1 year). PMID- 16242212 TI - Epidemiology of Acinetobacter baumannii in a French university hospital: are isolation precautions necessary, useful or non-essential? PMID- 16242213 TI - Pharmacokinetics, toxicity of nasal cilia and immunomodulating effects in Sprague Dawley rats following intranasal delivery of thymopentin with or without absorption enhancers. AB - Thymopentin (TP 5), a synthetic pentapeptide, has been used in clinic as a modulator for immnuodeficiencies through intramuscular administration. The objectives of this study was to investigate the pharmacokinetics using normal rats and toxicity of nasal cilia as well as immunomodulating effects using immunosuppression rats after intranasal delivery of thymopentin with or without an absorption enhancer. The absorption extent of fluorescein isothiocyanate (FITC) labeled TP 5 via nasal delivery at a single dose is significantly improved by incorporating sodium deoxycholate, Brij 35 and chitosan, respectively. FITC-TP 5 can also be absorbed to such an extent ranging from 15 to 28% after intranasal administration of FITC-TP 5 alone, FITC-TP 5 with sodium caprylate, or with bacitracin, respectively. After seven consecutive days multiple dosing, TP 5 formulation with sodium deoxycholate or Brij 35 caused apparently injury to nasal cilia, indicating these two enhancers would not be appropriate for nasal delivery. Results from superoxide dismutase activity, maleic dialdehyde, T lymphocyte subsets (CD3+, CD4+, CD8+ and CD4+/CD8+ ratio) analyses suggest that all the selected enhancers improve the modulating effects of TP 5 in the immunosuppression rats. On an overall evaluation, intranasal TP 5 alone, TP 5 with chitosan, or TP 5 with bacitracin formulation may be suitable for the future clinical application. PMID- 16242214 TI - Synthesis and activity of an octapeptide inhibitor designed for SARS coronavirus main proteinase. AB - The outbreak of SARS, a life-threatening disease, has spread over many countries around the world. So far there is no effective drug for the treatment of SARS. Stimulated by the binding mechanism of SARS-CoV Mpro with the octapeptide AVLQSGFR reported recently as well as the "Chou's distorted key" theory, we synthesized the octapeptide AVLQSGFR for conducting various biochemical experiments to investigate the antiviral potential of the octapeptide against SARS coronavirus (BJ-01). The results demonstrate that, compared with other compounds reported so far, AVLQSGFR is the most active in inhibiting replication of the SARS coronavirus, and that no detectable toxicity is observed on Vero cells under the condition of experimental concentration. PMID- 16242215 TI - Phosphorylation of the growth factors bFGF, NGF and BDNF: a prerequisite for their biological activity. AB - The aim of this work was to test whether growth factors such as basic fibroblast growth factor (bFGF), nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) undergo autophosphorylation and whether this affects their biological activity. Incubation of those growth factors with [gamma-(32)P]ATP resulted in phosphorylation in vitro. The phosphate bond was resistant to alkaline pH, yet acid-labile. Addition of alkaline phosphatase resulted in time and protein dependent dephosphorylation. Concomitantly, alkaline phosphatase abolished the neuroprotective effect of those growth factors upon oxygen and glucose deprivation and upon staurosporine-induced cell death. For those studies, we were using primary cultures of cortical and hippocampal neurons from embryonic and neonatal rats. Incubation of bFGF with non-hydrolyzable ATP-gammaS resulted in phosphorylation and in neuroprotection resistant to alkaline phosphatase. We conclude that bFGF, NGF and BDNF undergo autophosphorylation on site(s) other than serine, threonine, tyrosine and/or ATP-binding, and that this binding of phosphate is essential for neuroprotection in vivo. PMID- 16242217 TI - [Segmental and focal glomerulonephritis in the course of primitive Gougerot Sjogren syndrome]. PMID- 16242216 TI - [A thoracic tumor]. PMID- 16242218 TI - [Factitious Cushing syndrome: two case reports]. AB - INTRODUCTION: Search of corticosteroid use is an important precaution before any exploration of hypercorticism. Nevertheless, this use may be factitious leading to serious misdiagnosis and pitfalls. EXEGESIS: We report two cases of 29 and 60 years old male patients, referred for hypercorticism with patent features including metabolic and trophical signs. Laboratories findings revealed unelevated urinary cortisol values (40 and 27 microg/ day) in both cases with normal ACTH and normal plasmatic cortisol response to synacthene (32,5 and 35 microg/dl). Corticosteroid use, initially denied, was finally approved by the patients (hydrocortisone and dexamethasone for the first patient and betametasone for the second). So, they were assigned to psychiatric therapy. CONCLUSION: Our cases emphasize that corticosteroid misuse must be evoked in patients with unexpected hormonal assessment of pituitary adrenal axis despite patent clinical features of hypercorticism. Several specific tests are actually available for detect factitious Cushing syndrome. However, differential diagnostic remains difficult mainly with pseudo Cushing and cyclical Cushing syndromes. PMID- 16242219 TI - The effect of antigen targeting sequences on antibody responses to hepatitis E virus DNA vaccines in rats and sheep. AB - Expression of the capsid (PORF2) protein of hepatitis E virus (HEV) in mammalian cells results in heterogeneous intracellular processing with a mixture of stable and rapidly degraded forms, which might be expected to influence immune responses to DNA immunisation. Plasmids encoding the N-terminal 22 or 50 amino acids of PORF2 (Sig1 or Sig3, respectively) fused at the N-terminus of the ORF2.1 antigen of HEV (amino acids 394-660 of PORF2) were examined for processing in vitro and antibody responses in vivo, in both rats and sheep. Unmodified ORF2.1 is an unstable cytosolic protein and Sig1-ORF2.1 is a stable membrane-associated protein, whereas Sig3-ORF2.1 demonstrated heterogeneous processing analogous to that of full-length PORF2. After DNA immunisation, Sig1-ORF2.1 demonstrated a 30 fold enhancement of antibody responses in rats compared to untargeted ORF2.1, increasing to more than 200-fold after boosting with recombinant protein, but was ineffective in sheep. In contrast, Sig3-ORF2.1 did not give a significant effect in rats, but demonstrated 4-5-fold enhancement of antibody responses in sheep, and this enhancement was maintained after boosting with recombinant protein. These results suggest that Sig3 in particular may have promise as a targeting molecule for DNA vaccines in large animals. PMID- 16242220 TI - DNA vaccines in sheep: CTLA-4 mediated targeting and CpG motifs enhance immunogenicity in a DNA prime/protein boost strategy. AB - DNA vaccines have proven to be an efficient means of inducing immune responses in small laboratory animals; however, their efficacy in large out-bred animal models has been much less promising. In addressing this issue, we have investigated the ability of ovine cytotoxic lymphocyte antigen 4 (CTLA-4) mediated targeting and ruminant specific CpG optimised plasmids, both alone and in combination, to enhance immune responses in sheep to the pro cathepsin B (FhCatB) antigen from Fasciola hepatica. In this study, CTLA-4 mediated targeting enhanced the speed and magnitude of the primary antibody response and effectively primed for a potent memory response compared to conventional DNA vaccination alone, which failed to induce a detectable immune response. While the CpG-augmentation of the CTLA-4 targeted construct did not further enhance the magnitude or isotype profile of the CTLA-4 induced antibody titres, it did result in the induction of significant antigen-specific, lymphocyte-proliferative responses that were not observed in any other treatment group, showing for the first time that significant cellular responses can be induced in sheep following DNA vaccination. In contrast, CpG-augmentation in the absence of CTLA-4 mediated targeting failed to induce a detectable immune response. This is the first study to explore the potential adjuvant effects of ruminant specific CpG motifs on DNA vaccine induced immune responses in sheep. The ability of CpG-augmented CTLA-4 mediated targeting to induce both humoral and cellular immune responses in this study suggests that this may be an effective approach for enhancing the efficacy of DNA vaccines in large out-bred animal models. PMID- 16242221 TI - Safety and immunogenicity of New Zealand strain meningococcal serogroup B OMV vaccine in healthy adults: beginning of epidemic control. AB - As the first step towards control of a strain specific epidemic of meningococcal disease in New Zealand (NZ), this study, an observer-blind, randomised controlled trial in 75 healthy adults, evaluated safety and immunogenicity of two different dosages of a meningococcal group B vaccine administered in a three dose regime. The "tailor-made" outer membrane vesicle (OMV) vaccine (candidate vaccine) developed using a New Zealand meningococcal group B strain (B:4:P1.7b,4) was well tolerated with no vaccine related serious adverse events. Similar local and systemic reactions were observed in those receiving the New Zealand candidate vaccine and the control parent Norwegian vaccine (MenBvac). A four-fold rise in serum bactericidal antibodies (SBAb) against the vaccine strain 4-6 weeks after the third vaccination was achieved in 100% of New Zealand candidate vaccine 2,519 microg participants and in 87% of 50 microg participants. The safety and immunogenicity profile observed in this study of healthy adults enabled studies in children to be initiated using 25 microg dosage. PMID- 16242222 TI - Growth of the damselfly Ischnura heterosticta is better in saline water than freshwater. AB - Increasing salinity has the potential to affect freshwater organisms. Yet sub lethal effects of salinity on macroinvertebrates are poorly understood. Growth and development of Ischnura heterosticta (Odonata: Coenagrionidae) was experimentally shown to be faster in 5-20 mS/cm than 0.1-1 mS/cm, while in 35 mS/cm all individuals died. In 30 mS/cm about half died and growth was similar to the 0.1 mS/cm treatment. The salinity-growth relationship cannot be explained indirectly, that is salinity affecting the survival of their prey. Tissue content and concentration of Ca, Mg, Na and K in emerged adults showed no evidence of deficiencies at low salinity. Heart beat rate was similar across treatments, except at 35 mS/cm, where it was slower. Respiration and feeding were similar at 0.1, 10 and 30 mS/cm. While there are similarities in I. heterosticta and other species' salinity response, there are differences and studies on more species are urgently needed. PMID- 16242223 TI - Distributions and concentrations of PAHs in Hong Kong soils. AB - Surface soil (0-10 cm) samples from 53 sampling sites including rural and urban areas of Hong Kong were collected and analyzed for 16 EPA priority polycyclic aromatic hydrocarbons (PAHs). Total PAH concentrations were in the range of 7.0 410 microg kg(-1) (dry wt), with higher concentrations in urban soils than that in rural soils. The three predominant PAHs were Fluoranthene, Naphthalene and Pyrene in rural soils, while Fluoranthene, Naphthalene and Benzo(b + k)fluoranthene dominated the PAHs of urban soils. The values of PAHs isomer indicated that biomass burning might be the major origin of PAHs in rural soils, but vehicular emission around the heavy traffic roads might contribute to the soil PAHs in urban areas. A cluster analysis was performed and grouped the detectable PAHs under 4 clusters, which could be indicative of the PAHs with different origins and PAHs affected by soil organic carbon contents respectively. PMID- 16242224 TI - Linking organic pollutant (bio)availability with geosorbent properties and biomimetic methodology: a review of geosorbent characterisation and (bio)availability prediction. AB - The interdependent link between structure and physico-chemical properties of geosorbents and sorption activity of hydrophobic organic pollutants (HOC) upon interaction with solid matrices has been established. The conclusions derived from these investigations have not been actively incorporated into risk assessment and remediation protocols since legislators prefer to adopt a conservative approach when the potential of contaminants to be released from soil matrices are evaluated. With the advent of spectroscopic techniques, it is possible to determine the molecular properties of the geosorbent organic matter which play a pivotal role in HOC retention. Physical-chemical and biological methods are employed to predict the potential for HOC release from sorbent matrices. This article serves as a review discussing the literature and reports the progress that has been made in these particular areas. The implication of employing molecular descriptor factors correlated with a biomimetic method to assess availability and risk is also considered. PMID- 16242225 TI - Effect of humic acid on water chemistry, bioavailability and toxicity of aluminium in the freshwater snail, Lymnaea stagnalis, at neutral pH. AB - The influence of humic acid on the water chemistry of environmentally relevant concentrations of Al at neutral pH was studied, together with its effect on the bioavailability and toxicity of Al in Lymnaea stagnalis. Humic acid significantly reduced the loss of Al from the water and increased the fraction of filterable Al, although this was a relatively small fraction of total Al. Filterable Al concentration in the presence or absence of humic acid was independent of initial Al concentration. Humic acid only partly reduced toxicity, as observed by a reduction in behavioural suppression, and had no effect on the level of Al accumulated in tissues. These results suggest that humic acid maintains Al in a colloidal form that is bioavailable to L. stagnalis. However, these colloidal Al humic acid species were less toxic since behavioural toxicity was reduced. Humic acid may play an important role in limiting the toxicity of Al to freshwater organisms. PMID- 16242226 TI - In silico screening of chemicals for bacterial mutagenicity using electrotopological E-state indices and MDL QSAR software. AB - Quantitative structure-activity relationship (QSAR) software offers a rapid, cost effective means of prioritizing the mutagenic potential of chemicals. MDL QSAR models were developed using atom-type E-state indices and non-parametric discriminant analysis. Models were developed for Salmonella typhimurium gene mutation, combining results from strains TA97, TA98, TA100, TA1535, TA1536, TA1537, and TA1538 (n=3228), and Escherichia coli gene mutation tests WP2, WP100, and polA (n=472). Composite microbial mutation models (n=3338) were developed combining all Salmonella, E. coli, and the Bacillus subtilis rec spot test study results. The datasets contained 74% non-pharmaceuticals and 26% pharmaceuticals. Salmonella and microbial mutagenesis external validation studies included a total of 1444 and 1485 compounds, respectively. The average specificity, sensitivity, positive predictivity, concordance, and coverage of Salmonella models was 76, 81, 73, 78, and 98%, respectively, with similar performance for the microbial mutagenesis models. MDL QSAR and discriminant analysis provides rapid and highly automated mutagenicity screening software with good specificity, sensitivity, and coverage that is simpler and requires less user intervention than other similar software. MDL QSAR modules for microbial mutagenicity can provide efficient and cost effective large scale screening of compounds for mutagenic potential for the chemical and pharmaceutical industry. PMID- 16242227 TI - Our health and theirs: forced migration, othering, and public health. AB - This paper uses 'othering' theory to explore how forced migrants are received in developed countries and considers the implications of this for public health. It identifies a variety of mechanisms by which refugees, asylum seekers and irregular migrants are positioned as 'the other' and are defined and treated as separate, distant and disconnected from the host communities in receiving countries. The paper examines how this process has the potential to affect health outcomes both for individuals and communities and concludes that public health must engage with and challenge this othering discourse. It argues that public health practitioners have a critical role to play in reframing thinking about health services and health policies for forced migrants, by promoting inclusion and by helping shape a narrative which integrates and values the experiences of this population. PMID- 16242229 TI - Antimutagenic effect of polysaccharide ginsan extracted from Panax ginseng. AB - Ginsan is a polysaccharide extracted from the roots of Panax ginseng, and it has earlier been reported to have an immunostimulatory effect. In the present study, the frequency of micronucleated polychromatic erythrocytes (MNPCE) was assessed in the bone marrow of C57BL/6 male mice treated with ginsan [100, 200 or 300 mg/kg body weight (b.w.)] or amifostine (200mg/kg b.w.) 30 min before as well as 15 min after 1.5 Gy of gamma-irradiation. Ginsan and amifostine did not alter the frequency of MNPCE of control mice (P>0.05), showing that they are non-mutagenic per se; gamma-irradiation induced a statistically significant (P<0.001) increase of MNPCE and decrease of PCE/NCE ratio (P<0.001) compared to control group. However, ginsan applied 30 min before or 15 min after irradiation reduced MNPCE in a dose-dependent manner. Amifostine (200mg/kg b.w.) did not reduce radiation induced MNPCE, but stimulated erythropoiesis, when administered before irradiation. Based on the above results, radioprotective effect of ginsan can be partially attributed to reduction of radiation-induced genotoxicity. PMID- 16242230 TI - Differential cytotoxic responses of PC12 cells chronically exposed to psychostimulants or to hydrogen peroxide. AB - Repeated abuse of stimulant drugs, cocaine and amphetamine, is associated with extraneuronal dopamine accumulation in specific brain areas. Dopamine may be cytotoxic through the generation of reactive oxygen species, namely hydrogen peroxide (H2O2), resulting from dopamine oxidative metabolism. In this work, we studied the cytotoxicity in PC12 cells (a dopaminergic neuronal model) chronically and/or acutely exposed to cocaine or amphetamine, as compared to H2O2 exposure. Chronic cocaine treatment induced sensitization to acute cocaine insult and increased cocaine-evoked accumulation of extracellular dopamine, although no changes in dihydroxyphenylacetic acid (DOPAC) levels were observed. Moreover, dopamine was depleted in cells chronically exposed to amphetamine and acute amphetamine toxicity persisted in these cells, indicating that dopamine was not involved in amphetamine cytotoxicity. PC12 cells chronically treated with H2O2 were totally resistant to acute H2O2, but not to acute cocaine or amphetamine exposure, suggesting that the toxicity induced by these stimulant drugs is unrelated to adaptation to oxidative stress. Interestingly, chronic cocaine treatment largely, but not completely, protected the cells against a H2O2 challenge, whilst a decrement in intracellular ATP was observed. This study shows that chronic treatment of PC12 cells with cocaine or H2O2 modifies the cytotoxic response to an acute exposure to these agents. PMID- 16242231 TI - Effect of subchronic exposure to acrylamide induced on the expression of bcl-2, bax and caspase-3 in the rat nervous system. AB - Occupational exposure and experimental intoxication with acrylamide (ACR) produce a neuropathy characterized by nerve degeneration. We hypothesize that ACR differentially affects the expression of bcl-2, bax and caspase-3 in the central nervous system (CNS) and the peripheral nervous system (PNS) tissue. Male adult Wistar rats were given ACR (20, 40 mg/kg i.p. 3 days/week) for 8 weeks. Samples of the cerebral cortex, cerebellum, spinal cord and sciatic nerves were collected and examined for bcl-2, bax and caspase-3 expression using Western blotting. Subchronic exposure to ACR reduced cortical bcl-2 expression in the low dose, increased it in the high dose; the change of bcl-2 expression in the spinal cord and cerebellum followed the same pattern as that described in the cerebral cortex; there was no significant change in the expression of bax in the cerebral cortex and the spinal cord, however, in the cerebellum the change of bax expression and bcl-2 expression is just the reverse. Thus, the bcl-2/bax ratio of the CNS tissue was affected by exposure to ACR, it decreased in the low dose group and increased in the high group. Compared to control, densitometric analysis showed that in the sciatic nerves the expression of bcl-2 and bax expression was markedly increased following ACR administration. The expression of inactive isoforms (32 kDa) of caspase-3 was not altered in the cortices of ACR treated rats, but increased in their spinal cords and sciatic nerves. Thus, subchronic exposure to ACR affected the expression of death-related proteins in the CNS and PNS tissue, which indicate there is the early molecular regulatory mechanism of apoptosis in the neuropathy induced by ACR. PMID- 16242233 TI - Solvent effects on the reactivity of fluorenyl nitrenium ion with DNA-like probes. AB - Laser Flash Photolysis (LFP) experiments carried out on 2-azidofluorene in aqueous systems generate 2-fluorenyl nitrenium ion (lambda(max) = 450 nm) which decays with first order rate constant and is quenched with 2'-deoxyguanosine originating an intermediate, namely the C8 adduct of 2-fluorenyl nitrenium ion, with bimolecular rate constant in the order of 1.3 x 10(9) M(-1) s(-1) in pure water. This intermediate very likely mimics the intermediate formed from carcinogens (i.e.: arylnitrenium ions formed through metabolic activation pathways from aminoaromatic substrates) and DNA rests in vivo. Solvent effects demonstrate and support the further stabilization of this intermediate (with respect to fluorenyl nitrenium ion) through hydrogen bonding as compared to other probe systems, and accounts for the enhanced metabolic carcinogenecity observed for this type of compounds. Diverse solvent systems, such as mixtures of water with acetonitrile, 1,1,1-trifluoroethanol, and 1,1,1,3,3,3-hexafluoroisopropanol, are used to interpret solvent-complex interactions. PMID- 16242232 TI - Clinical evaluation of composite and compomer restorations in primary teeth: 24 month results. AB - PURPOSE: This split-mouth, blind study evaluated the clinical performance of Dyract AP, F2000, and Heliomolar placed in primary molars of 30 children (mean age, 6 years and 2 months). MATERIALS AND METHODS: From a total of 79 restorations accomplished, 27 were built with Heliomolar (18 Class I, and 9 Class II), 30 were with F2000 (21 Class I and 9 Class II), and 22 were built with Dyract AP (14 Class I and 8 Class II). All of teeth restored had primary caries lesions. At 6, 12, 18 and 24 months, 60 restorations (75%) were evaluated using USPHS criteria for: color match (CM), marginal adaptation (MA), marginal discoloration (MD), anatomic form (AF) and secondary caries (SC) by three calibrated operators. The Alpha+Bravo score percentage was considered as clinical success. The data were subjected to statistical analysis by Mann-Whitney and Kruskal-Wallis tests (p<0.05). RESULTS: Heliomolar showed the smallest success clinical at 12 months for marginal adaptation and secondary caries, and at 18 months for marginal discoloration. Regarding color match and anatomic form, no significant differences were found among the groups at each evaluation period. When materials were compared, Heliomolar did not show a significant difference among the evaluated periods for any criteria, remaining with the lowest scores. Significant differences were observed at 12 months for F2000 (marginal adaptation), and at 24 months for Dyract AP (marginal discoloration and secondary caries) and for F2000 (color match and marginal discoloration). CONCLUSIONS: It was concluded that Dyract AP and F2000 showed the best clinical performance over 24 month-evaluations for marginal discoloration and secondary caries, and color match and marginal adaptation, respectively. The use of the resin composite Heliomolar in Class I/II restorations in primary molars should be carefully considered. PMID- 16242234 TI - Binding and photocleavage of cationic porphyrin-phenylpiperazine hybrids to DNA. AB - The binding properties of cationic porphyrin-phenylpiperazine hybrids to calf thymus (CT) DNA were investigated by using absorption, fluorescence and circular dichroism (CD) spectra, and the apparent affinity binding constants (K(app)) of the porphyrins for CT DNA were determined by using a competition method with ethidium bromide (EB). Intercalation of porphyrin into CT DNA occurred when two phenylpiperazines were introduced at cis position onto the periphery of cationic porphyrin. The photocleavages of pBR322 plasmid DNA by the porphyrins were consistent with the values of K(app). With [porphyrin]/[DNA base pairs] ratio increased, the binding mode tended to be outside binding, and the cleavage abilities of the porphyrins varied. In the presence of sodium azide, a quencher of 1O2, the cleavage of DNA by the porphyrin of intercalation was less inhibited. PMID- 16242236 TI - Towards building the silicon cell: a modular approach. AB - Systems Biology aims to understand quantitatively how properties of biological systems can be understood as functions of the characteristics of, and interactions between their macromolecular components. Whereas, traditional biochemistry focused on isolation and characterization of cellular components, the challenge for Systems Biology lies in integration of this knowledge and the knowledge about molecular interactions. Computer models play an important role in this integration. We here discuss an approach with which we aim to link kinetic models on small parts of metabolism together, so as to form detailed kinetic models of larger chunks of metabolism, and ultimately of the entire living cell. Specifically, we will discuss techniques that can be used to model a sub-network in isolation of a larger network of which it is a part, while still maintaining the dynamics of the larger complete network. We will start by outlining the JWS online system, the silicon cell project, and the type of models we propose. JWS online is a model repository, which can be used for the storage, simulation and analysis of kinetic models. We advocate to integrate a top-down approach, where measurements on the complete system are used to derive fluxes in a detailed structural model, with a bottom-up approach, consisting of the integration of molecular mechanism-based detailed kinetic models into the structural model. PMID- 16242237 TI - Generative topographic mapping applied to clustering and visualization of motor unit action potentials. AB - The identification and visualization of clusters formed by motor unit action potentials (MUAPs) is an essential step in investigations seeking to explain the control of the neuromuscular system. This work introduces the generative topographic mapping (GTM), a novel machine learning tool, for clustering of MUAPs, and also it extends the GTM technique to provide a way of visualizing MUAPs. The performance of GTM was compared to that of three other clustering methods: the self-organizing map (SOM), a Gaussian mixture model (GMM), and the neural-gas network (NGN). The results, based on the study of experimental MUAPs, showed that the rate of success of both GTM and SOM outperformed that of GMM and NGN, and also that GTM may in practice be used as a principled alternative to the SOM in the study of MUAPs. A visualization tool, which we called GTM grid, was devised for visualization of MUAPs lying in a high-dimensional space. The visualization provided by the GTM grid was compared to that obtained from principal component analysis (PCA). PMID- 16242235 TI - Trading the micro-world of combinatorial complexity for the macro-world of protein interaction domains. AB - Membrane receptors and proteins involved in signal transduction display numerous binding domains and operate as molecular scaffolds generating a variety of parallel reactions and protein complexes. The resulting combinatorial explosion of the number of feasible chemical species and, hence, different states of a network greatly impedes mechanistic modeling of signaling systems. Here we present novel general principles and identify kinetic requirements that allow us to replace a mechanistic picture of all possible micro-states and transitions by a macro-description of states of separate binding sites of network proteins. This domain-oriented approach dramatically reduces computational models of cellular signaling networks by dissecting mechanistic trajectories into the dynamics of macro- and meso-variables. We specify the conditions when the temporal dynamics of micro-states can be exactly or approximately expressed in terms of the product of the relative concentrations of separate domains. We prove that our macro modeling approach equally applies to signaling systems with low population levels, analyzed by stochastic rather than deterministic equations. Thus, our results greatly facilitate quantitative analysis and computational modeling of multi-protein signaling networks. PMID- 16242238 TI - Ultrasound facilitates and improves removal of Cd(II) from aqueous solution by the discarded tire rubber. AB - Some of the heavy metal ions such as cadmium are toxic and represent as hazardous pollutants due to their persistence in the environment. In this study the ground discarded tire rubber was used for the sorption of cadmium from aqueous solution. The batch sorption tests were conducted to investigate the sorption of Cd(II) by discarded tire rubber in the presence and absence of ultrasound. To assess the capability of sorbent, research parameters such as ultrasonic waves, solution temperature, particle size of ground tire and others were investigated. The experimental data were fitted in Langmuir model better than Freundlich one. Therefore, the former model was applied to the sorption equilibrium in order to determine the maximum metal sorption capacity in the presence and absence of ultrasound. The Langmuir constants were also obtained from the isotherms under different conditions. In the presence of ultrasound the tire rubber was a more efficient sorbent for this pollutant than its absence. According to the results, the internal porous and film diffusions were both effective in the sorption process. The porous and film diffusion coefficients of the ground tire rubber were, respectively, about 1.8 and 2.7 times more in the presence of ultrasound than its absence. The effect of ultrasound on the sorption process could be explained by the thermal and non-thermal properties of ultrasonic field. PMID- 16242239 TI - Pentylenetetrazole kindling induces neuronal cyclin B1 expression in rat hippocampus. AB - The purpose of the study was to explore the involvement of cell cycle events in the neuronal death induced by repeated seizures. Pentylenetetrazole (PTZ) kindling was used as a model of seizure-induced hippocampal neurodegeneration. Immunohistochemical approach was applied to detect cell cycle markers (cyclins and cycline-dependent kinases) in hippocampus. PTZ-kindling in rats induced moderate neuronal cell loss in hippocampal fields CA1, CA 3, CA 4, and dentate gyrus. The majority of damaged cells in hippocampi of PTZ-kindled rats were cycline B1 positive, while no expression of either other cell cycle markers or TUNEL-positive (apoptotic) nuclei could be revealed. Since cycline B1 expression has been described in hippocampal neurons of patients with temporal lobe epilepsy by [Z. Nagy, M.M. Esiri, Neuronal cyclin expression in the hippocampus in temporal lobe epilepsy, Exp. Neurol. 150 (1998) 240-247], it is suggested that PTZ-kindling may be a suitable model to study the mechanisms of seizure-induced neuronal death. PMID- 16242240 TI - Neurotrophic factors increase tumor necrosis factor-alpha-induced nuclear translocation of NF-kappaB in rat PC12 cells. AB - Neurotrophic factors regulate neuronal survival and differentiation and control neurite outgrowth by binding to tyrosine kinase receptors, the Trks, and a tumor necrosis factor (TNF) receptor-like molecule, p75 neurotrophin receptor. A proinflammatory cytokine, TNF, also affects survival and apoptotic death in neuronal cells. However, it is still unclear whether neurotrophic factors and TNF co-operate the intracellular signaling. Using green fluorescent protein-tagged NF kappaB1 (GFP-NF-kappaB1), we examined here the effects of TNF-alpha and neurotrophic factors on the nuclear translocation of NF-kappaB in PC12 cells. TNF alpha induced gradually the translocation of GFP-NF-kappaB1 from the cytoplasm to the nucleus within 60 min. Pretreatment of lactacystin which is a proteasome specific inhibitor suppressed significantly the nuclear translocation of GFP-NF kappaB1 after TNF-alpha stimulation. In addition, we found that co-stimulation of TNF-alpha and neurotrophic factors such as nerve growth factor (NGF) and brain derived neurotrophic factor (BDNF) increased greatly the nuclear translocation of GFP-NF-kappaB1 whereas neither NGF nor BDNF itself induced the translocation. These results suggested that there is a close correlation between the signaling pathways via TNF receptors and neurotrophin receptors for the NF-kappaB activation, and that NGF and BDNF enhance TNF-alpha-induced nuclear translocation of NF-kappaB. PMID- 16242242 TI - Adding temozolomide to radiotherapy prolongs survival in people with glioblastoma. PMID- 16242241 TI - Inhibition of SOD1 expression by mitomycin C is a non-specific consequence of cellular toxicity. AB - Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative condition that results in the death of the large motor neurons of the brain and spinal cord. Familial ALS accounts for 10% of all ALS cases. Approximately 25% of these cases are due to mutations in the SOD1 gene. Several lines of evidence argue that mutant SOD1 causes ALS by a toxic gain of function. We therefore anticipate that measures that reduce the levels of mutant SOD1 expression should be beneficial in mutant SOD1-associated ALS patients. Mitomycin C (MC) is an antitumor antibiotic previously demonstrated to reduce SOD1 expression in a reporter gene system. We investigated whether MC reduces endogenous SOD1 expression levels both in vitro and in vivo. MC reduced human and rat SOD1 protein levels in vitro, with a concomitant decrease in actin and increase in p53 protein levels, as detected by Western blotting. However, this decrease in SOD1 protein levels was paralleled by a similar decrease in cell viability. In contrast, intracerebroventricular administration of MC to rats and mice failed to produce any effect on brain or spinal cord SOD1 protein levels. Our data indicate the apparent inhibition of SOD1 expression by MC is a non-specific consequence of MC-induced cellular toxicity. PMID- 16242243 TI - Gemcitabine plus docetaxel for advanced or metastatic non-small cell lung cancer: similar survival to cisplatin plus vinorelbine and less toxicity. PMID- 16242244 TI - Impulsivity, inhibition and negative priming in ecstasy users. AB - A modified Stroop color-word interference paradigm was used to investigate the effects of recreational ecstasy (MDMA) use on central executive inhibitory processes. Ecstasy users who also used cannabis were compared with non-users matched for cannabis consumption and with non-drug users on a Stroop task in which standard color-word interference trials were interspersed with trials in which the target color was the same as the distractor word on the immediately preceding trial. Ecstasy's effects on standard inhibition (conscious suppression of a prepotent response pattern--responsible for Stroop interference) could thus be contrasted with its effect on the short-term, unconscious, inhibitory process responsible for suppression of the preceding distractor word (negative priming). Neither drug group differed from the non-drug users in level of Stroop interference but ecstasy users showed reduced negative priming compared to the cannabis users and non-drug users. The loss of inhibition in the ecstasy users was not related to impulsivity assessed by two standard scales since these were similar in both drug-user groups and raised relative to the non-drug users. It is argued that previous failures to demonstrate loss of inhibition could be partly due to the fact that standard executive function tests, such as the Stroop, are unable to differentiate between sub-types of inhibition, only some of which may be affected by ecstasy use. PMID- 16242245 TI - Transtheoretical model constructs in smokers with and without medical illness: a second look at the medical effect. AB - The present report describes an investigation of differences in transtheoretical model constructs between 275 smokers with any medical illness, smokers with chronic illness specifically, and smokers who are medically healthy. In contrast to a previous report by Kristeller et al., we did not find higher process scores among medically ill smokers, instead finding more relapses and higher temptation scores among the medically ill smokers. Chronically ill smokers in particular reported high temptation to smoke in negative affect and habit/craving situations. Moreover, greater number of chronic illnesses was associated with increasing temptation and pros of smoking. We conclude that while medical smokers may report more quit attempts, they may have more difficulty staying quit than their healthy counterparts. Helping medically ill smoking patients to cope more effectively with highly tempting situations, to decrease perceived benefits of smoking, and to increase their use of the processes of change may lead to greater cessation success. PMID- 16242246 TI - Predictors of smoking relapse in a sample of Dutch adult smokers; the roles of gender and action plans. AB - BACKGROUND: The object of this study was to investigate the influence of gender and the additional protective quality of action plans on the occurrence of relapse in smokers after a pharmacy-based minimal intervention strategy. METHODS: Respondents received a written baseline questionnaire, based on the I-Change Model, and were contacted 3 months after baseline by telephone. RESULTS: A total of 223 respondents were included in relapse analyses. Relapse was predicted by intention, the use of pharmacotherapy and action plans. Predictors differed for men and women. CONCLUSIONS: Action plans provide a protective value against smoking relapse. Men and women did not differ in relapse rates, but predictors differed. Developers of relapse prevention activities should consider developing programmes based on gender. These programmes could also benefit from making use of action plans as a method for preventing relapse. PMID- 16242247 TI - Sublethal mitochondrial stress with an attendant stoichiometric augmentation of reactive oxygen species may precipitate many of the beneficial alterations in cellular physiology produced by caloric restriction, intermittent fasting, exercise and dietary phytonutrients: "Mitohormesis" for health and vitality. AB - The precise mechanistic sequence producing the beneficial effects on health and lifespan seen with interventions as diverse as caloric restriction, intermittent fasting, exercise, and consumption of dietary phytonutrients is still under active characterization, with large swaths of the research community kept in relative isolation from one another. Among the explanatory models capable of assisting in the identification of precipitating elements responsible for beneficial influences on physiology seen in these states, the hormesis perspective on biological systems under stress has yielded considerable insight into likely evolutionarily consistent organizing principles functioning in all four conditions. Recent experimental findings provide the tantalizing initial lodestones for an entirely new research front examining molecular substrates of stress resistance. In this novel body of research, a surprising new twist has emerged: Reactive oxygen species, derived from the mitochondrial electron transport system, may be necessary triggering elements for a sequence of events that result in benefits ranging from the transiently cytoprotective to organismal level longevity. With the recent appreciation that reactive oxygen species and reactive nitrogen species function as signaling elements in a interconnected matrix of signal transduction, the entire basis of many widely accepted theories of aging that predominated in the past may need to be reconsidered to facilitate the formulation of an new perspective more correctly informed by the most contemporaneous experimental findings. This perspective, the mitohormesis theory, can be used in many disparate domains of inquiry to potentially explain previous findings, as well as point to new targets of research. The utility of this perspective for research on aging is significant, but beyond that this perspective emphasizes the pressing need to rigorously characterize the specific contribution of the stoichiometry of reactive oxygen species and reactive nitrogen species in the various compartments of the cell to cytoprotection and vitality. Previous findings regarding the influences of free radical chemistry on cellular physiology may have represented assessments examining the consequences of isolated elevation of signaling elements within a larger signal transductive apparatus, rather than definitive characterizations of the only modality of reactive oxygen species (and reactive nitrogen species) influence. In applying this perspective, it may be necessary for the research community, as well as the practicing clinician, to engender a more sanguine perspective on organelle level physiology, as it is now plausible that such entities have an evolutionarily orchestrated capacity to self-regulate that may be pathologically disturbed by overzealous use of antioxidants, particularly in the healthy. PMID- 16242248 TI - Inactivation of prion proteins via covalent grafting with methoxypoly(ethylene glycol). AB - Transmissible spongiform encephalopathies (TSE) such as bovine spongiform encephalitis (BSE), Creutzfeld-Jakob disease (CJD) as well as other proteinaceous infectious particles (prions) mediated diseases have emerged as a significant concern in transfusion medicine. This concern is derived from both the disease causing potential of prion contaminated blood products but also due to tremendous impact of the active deferral of current and potential blood donors due to their extended stays in BSE prevalent countries (e.g., the United Kingdom). To date, there are no effective means by which infectious prion proteins can be inactivated in cellular and acellular blood products. Based on current work on the covalent grafting of methoxypoly(ethylene glycol) [mPEG] to proteins, viruses, and anuclear, and nucleated cells, it is hypothesized that the conversion of the normal PrP protein to its mutant conformation can be prevented by the covalent grafting of mPEG to the mutant protein. Inactivation of infective protein particles (prions) in both cellular blood products as well as cell free solutions (e.g., clotting factors) could be of medical/commercial value. It is hypothesized that consequent to the covalent modification of donor-derived prions with mPEG the requisite nucleation of the normal and mutant PrP proteins is inhibited due to the increased solubility of the modified mutant PrP and that the conformational conversion arising from the mutant PrP is prevented due to obscuration of protein charge by the heavily hydrated and neutral mPEG polymers, as well as by direct steric hindrance of the interaction due to the highly mobile polymer graft. PMID- 16242249 TI - Boom or bubble? Is medical research thriving or about to crash? AB - A recent issue of JAMA (2005; vol. 294) presented a portrait of medical research as a booming enterprise. By contrast I have suggested that medical research is a speculative bubble due to burst. How can two such different predictions be compatible? From inside the expanding world of medical research everything seems fine and getting better. But to people outside the system, it seems like there is an awful lot of money going in, and not much coming out. Professional criteria of success (publications, impact factors, citations, grant income, large teams, etc.) are not the same as the outsider's view of success. Outsiders want the medical research system to generate therapeutic progress as efficiently as possible: the most progress for the least resources. Medical research is not the only good way of spending money and is in competition with other social systems. As funding increases, diminishing returns will set-in, opportunity costs will begin to bite, and there will be more and more social benefit to be gained from spending the extra research money on something else. Therefore, future cuts in medical research will happen because of pressure from outside the system - specifically pressure from other powerful social systems which will press their alternative claims for funding. In the short term, there will be a quantitative decline of research production. But in the longer term the medical research system will re-grow in a more efficient form. After a 'golden age of therapeutic progress in the mid-20th century, recent decades have seen a 'silver age' of scholasticism which is due to end soon. Perhaps a renaissance of medical research lies not too many years in the future. PMID- 16242250 TI - Senile plaques in Alzheimer's diseased brains: possible association of beta amyloid with herpes simplex virus type 1 (HSV-1) L-particles. AB - The characteristic insoluble, senile (neuritic) plaques found extracellularly in brains of patients with Alzheimer's disease (AD) contain the fibrillar form of beta-amyloid (Abeta42). A substantial proportion of autopsied elderly brains have demonstrated DNA evidence of herpes simplex virus type 1 (HSV-1) infiltration. HSV-1-infected cells produce significant quantities of non-infectious, non-DNA containing light particles (L-particles) comprised of viral envelope and tegument proteins. HSV-induced L-particles can be exocytosed out of their host cells. This report advances the hypothesis that (1) Abeta binds to L-particles; (2) Abeta permeabilizes L-particles, destroying the integrity of the envelope and allowing the contained tegument proteins to spill into the extracellular space; and (3) these events are followed by a conformational shift of Abeta into its fibrillar form, physically trapping the L-particle-derived substances and resulting in the plaques characteristic of AD. These hypotheses are supported by reports of biomolecular changes and pathophysiologies which have been simultaneously observed in both AD- and HSV-infected brains. PMID- 16242251 TI - [F-18]-fluorodeoxyglucose positron emission tomography for targeting radiation dose escalation for patients with glioblastoma multiforme: clinical outcomes and patterns of failure. AB - PURPOSE: [F-18]-fluorodeoxyglucose positron emission tomography (FDG-PET) imaging for brain tumors has been shown to identify areas of active disease. Radiation dose escalation in the treatment of glioblastoma multiforme may lead to improved disease control. Based on these premises, we initiated a prospective study of FDG PET for the treatment planning of radiation dose escalation for the treatment of glioblastoma multiforme. METHODS AND MATERIALS: Forty patients were enrolled. Patients were treated with standard conformal fractionated radiotherapy with volumes defined by MRI imaging. When patients reached a dose of 45-50.4 Gy, they underwent FDG-PET imaging for boost target delineation, for an additional 20 Gy (2 Gy per fraction) to a total dose of 79.4 Gy (n = 30). RESULTS: The estimated 1 year and 2-year overall survival (OS) for the entire group was 70% and 17%, respectively, with a median overall survival of 70 weeks. The estimated 1-year and 2-year progression-free survival (PFS) was 18% and 3%, respectively, with a median of 24 weeks. No significant improvements in OS or PFS were observed for the study group in comparison to institutional historical controls. CONCLUSIONS: Radiation dose escalation to 79.4 Gy based on FDG-PET imaging demonstrated no improvement in OS or PFS. This study establishes the feasibility of integrating PET metabolic imaging into radiotherapy treatment planning. PMID- 16242252 TI - Preoperative radiation with concurrent chemotherapy for resectable rectal cancer: effect of dose escalation on pathologic complete response, local recurrence-free survival, disease-free survival, and overall survival. AB - PURPOSE: Three Phase II studies of preoperative radiotherapy and concurrent 5FU chemotherapy were undertaken. The primary endpoints were acute toxicity and pathologic complete response rate (pCR). Secondary endpoints were local recurrence-free survival (LRFS), disease-free survival (DFS), and overall survival (OS). METHODS AND MATERIALS: A total of 134 patients with adenocarcinoma of the rectum (clinical T3/T4 or N1/N2) were treated. The initial cohort received 40 Gy in 20 fractions, the second 46 Gy in 23 fractions, and the third 50 Gy in 25 fractions. 5FU (225 mg/m2/day) was given continuously throughout radiotherapy. A total of 121 patients underwent surgical resection. RESULTS: Treatment was well tolerated. Grade 3/4 acute toxicity was observed in 13%, 4%, and 14% of patients in the 40 Gy, 46 Gy, and 50 Gy cohorts, respectively (p = 0.20). pCR was documented in 15%, 23%, and 33% of patients, respectively (p = 0.07). The 2-year actuarial LRFS was 72%, 90%, and 89% (p = 0.02); DFS was 62%, 84%, and 78% (p = 0.02); and OS was 72%, 94%, and 92%, respectively (p = 0.03). CONCLUSIONS: All treatment schedules were well tolerated. There was a trend toward increased pCR with higher doses. A statistically significant increase in LRFS, DFS, and OS was seen with radiation doses of 46 Gy and greater, but there was no difference between 46 Gy and 50 Gy. PMID- 16242253 TI - Hypoxia in relation to vasculature and proliferation in liver metastases in patients with colorectal cancer. AB - PURPOSE: To investigate hypoxia measured by pimonidazole binding, glucose transporter 1 (GLUT1) and carbonic anhydrase IX (CA-IX) expression, proliferation, and vascularity in liver metastases of colorectal cancer and to compare GLUT1 and CA-IX expression in corresponding primary tumors. METHODS AND MATERIALS: Twenty-five patients with liver metastases of colorectal cancer, planned for metastasectomy, were included. The hypoxia marker pimonidazole and proliferation marker iododeoxyuridine were administered before surgery. After immunofluorescent staining of the frozen metastases, pimonidazole binding, vascularity, and proliferation were analyzed quantitatively. Thirteen paraffin embedded primary tumors were stained immunohistochemically for GLUT1 and CA-IX expression, which was analyzed semiquantitatively in primary tumors and corresponding liver metastases. RESULTS: In liver metastases, pimonidazole binding showed a pattern consistent with diffusion-limited hypoxia. The mean pimonidazole-positive fraction was 0.146; the mean distance from vessels to pimonidazole-positive areas was 80 microm. When expressed, often co-localization was observed between pimonidazole binding and GLUT1 or CA-IX expression, but microregional areas of mismatch were also observed. No correlation between the level of pimonidazole binding and GLUT1 or CA-IX expression was observed. In some patients, a large fraction (up to 30%) of proliferating cells was present in pimonidazole-stained areas. Expression of CA-IX in primary tumors and metastases showed a significant correlation, which was absent for GLUT1 expression. CONCLUSIONS: Compared with other tumor types, liver metastases of colorectal cancer contain large amounts of hypoxic cells. The lack of correlation with pimonidazole binding brings into question the value of GLUT1 and CA-IX as endogenous markers of hypoxia. PMID- 16242254 TI - Chemoradiation and brachytherapy in biliary tract carcinoma: long-term results. AB - PURPOSE: To evaluate long-term effects of chemoradiation and intraluminal brachytherapy in terms of local control, disease-free survival, overall survival, and symptom relief in patients with unresectable or residual extrahepatic biliary carcinoma. METHODS AND MATERIALS: Twenty-two patients with unresectable (17 patients) or residual (5 patients) nonmetastatic extrahepatic bile tumors received external beam radiation therapy (39.6-50.4 Gy) between 1991 and 1997. In 21 patients, 5-fluorouracil (96-h continuous infusion, Days 1-4, 1,000 mg/m2/day) was administered. Twelve patients received a boost of intraluminal brachytherapy with 192Ir wires (30-50 Gy) 1 cm from the source axis. RESULTS: During external beam radiotherapy, 10 patients (45.4%) developed Grade 1 to 2 gastrointestinal toxicity. In patients with unresectable tumor who could be evaluated, the clinical response was 28.6% (4 of 14). Two patients showed complete response. In all 22 patients, median durations of local control, disease-free survival, and overall survival were 44.5 months, 16.3 months, and 23.0 months, respectively. Two patients who received external beam radiation therapy and intraluminal brachytherapy developed late duodenal ulceration. In patients with unresectable tumors, median survival was 13.0 months and 22.0 months in those treated with and without brachytherapy, with 16.7% and no 5-year survival, respectively (p=0.607). Overall 5-year survival was 18.0%: 40% and 11.7% in patients treated with partial resection and in those with unresectable tumor, respectively (p=0.135). CONCLUSION: This study confirmed the role of concurrent chemoradiation in advanced biliary carcinoma; the role of intraluminal brachytherapy boost remains to be further analyzed in larger clinical trials. PMID- 16242255 TI - Polymorphism at the 3'-UTR of the thymidylate synthase gene: a potential predictor for outcomes in Caucasian patients with esophageal adenocarcinoma treated with preoperative chemoradiation. AB - PURPOSE: To test the hypothesis that TS3'UTR polymorphisms predict outcomes in 146 Caucasian patients with esophageal adenocarcinoma treated with preoperative 5 fluorouracil-based chemoradiation. METHODS AND MATERIALS: DNA was extracted from hematoxylin-and-eosin stained histologic slides of normal esophageal or gastric mucosa sections from paraffin blocks of esophagectomy specimens. Genotypes of the TS3'UTR polymorphism were determined by polymerase chain reaction for a 6-bp insertion. The genotype groups (0bp/0bp, 6bp/0bp, and 6bp/6bp) were compared for clinical features and overall survival, recurrence-free-survival, locoregional control (LRC), and distant metastasis control. Multivariable Cox regression analyses were performed to find independent predictors for the stated outcomes. RESULTS: There was a trend of association between 6bp/6bp genotype and a decreased risk of local regional recurrence (hazards ratio = 0.211, 95% confidence interval = 0.041-1.095, p = 0.06) compared with other genotypes. There was a trend that patients with 6bp/6bp genotype had a higher 3-year probability of LRC compared with patients with the other two genotypes combined (p = 0.07); however, the difference was not statistically significant. CONCLUSIONS: The null hypotheses were not rejected in this study, probably owing to small sample size or the single gene examined. Prospective studies with adequate statistical power analyzing a family of genes involved in the 5-fluorouracil metabolism are needed to assess genetic determinants of treatment-related outcomes in esophageal adenocarcinoma. PMID- 16242256 TI - Dosimetry and preliminary acute toxicity in the first 100 men treated for prostate cancer on a randomized hypofractionation dose escalation trial. AB - PURPOSE: The alpha/beta ratio for prostate cancer is postulated to be between 1 and 3, giving rise to the hypothesis that there may be a therapeutic advantage to hypofractionation. The dosimetry and acute toxicity are described in the first 100 men enrolled in a randomized trial. PATIENTS AND METHODS: The trial compares 76 Gy in 38 fractions (Arm I) to 70.2 Gy in 26 fractions (Arm II) using intensity modulated radiotherapy. The planning target volume (PTV) margins in Arms I and II were 5 mm and 3 mm posteriorly and 8 mm and 7 mm in all other dimensions. The PTV D95% was at least the prescription dose. RESULTS: The mean PTV doses for Arms I and II were 81.1 and 73.8 Gy. There were no differences in overall maximum acute gastrointestinal (GI) or genitourinary (GU) toxicity acutely. However, there was a slight but significant increase in Arm II GI toxicity during Weeks 2, 3, and 4. In multivariate analyses, only the combined rectal DVH parameter of V65 Gy/V50 Gy was significant for GI toxicity and the bladder volume for GU toxicity. CONCLUSION: Hypofractionation at 2.7 Gy per fraction to 70.2 Gy was well tolerated acutely using the planning conditions described. PMID- 16242257 TI - Investigation of clinical and dosimetric factors associated with postoperative pulmonary complications in esophageal cancer patients treated with concurrent chemoradiotherapy followed by surgery. AB - PURPOSE: To assess the association of clinical and especially dosimetric factors with the incidence of postoperative pulmonary complications among esophageal cancer patients treated with concurrent chemoradiation therapy followed by surgery. METHOD AND MATERIALS: Data from 110 esophageal cancer patients treated between January 1998 and December 2003 were analyzed retrospectively. All patients received concurrent chemoradiotherapy followed by surgery; 72 patients also received irinotecan-based induction chemotherapy. Concurrent chemotherapy was 5-fluorouracil-based and in 97 cases included taxanes. Radiotherapy was delivered to a total dose of 41.4-50.4 Gy at 1.8-2.0 Gy per fraction with a three dimensional conformal technique. Surgery (three-field, Ivor-Lewis, or transhiatal esophagectomy) was performed 27-123 days (median, 45 days) after completion of radiotherapy. The following dosimetric parameters were generated from the dose volume histogram (DVH) for total lung: lung volume, mean dose to lung, relative and absolute volumes of lung receiving more than a threshold dose (relative V(dose) and absolute V(dose)), and absolute volume of lung receiving less than a threshold dose (volume spared, or VS(dose)). Occurrence of postoperative pulmonary complications, defined as pneumonia or acute respiratory distress syndrome (ARDS) within 30 days after surgery, was the endpoint for all analyses. Fisher's exact test was used to investigate the relationship between categorical factors and incidence of postoperative pulmonary complications. Logistic analysis was used to analyze the relationship between continuous factors (e.g., V(dose) or VS(dose)) and complication rate. Logistic regression with forward stepwise inclusion of factors was used to perform multivariate analysis of those factors having univariate significance (p < 0.05). The Mann-Whitney test was used to compare length of hospital stay in patients with and without lung complications and to compare lung volumes, VS5 values, and absolute and relative V5 values in male vs. female patients. Pearson correlation analysis was used to determine correlations between dosimetric factors. RESULTS: Eighteen (16.4%) of the 110 patients developed postoperative pulmonary complications. Two of these died of progressive pneumonia. Hospitalizations were significantly longer for patients with postoperative pulmonary complications than for those without (median, 15 days vs. 11 days, p = 0.003). On univariate analysis, female gender (p = 0.017), higher mean lung dose (p = 0.036), higher relative volume of lung receiving > or = 5 Gy (V5) (p = 0.023), and smaller volumes of lung spared from doses > or = 5 35 Gy (VS5-VS35) (p < 0.05) were all significantly associated with an increased incidence of postoperative pulmonary complications. No other clinical factors were significantly associated with the incidence of postoperative pulmonary complications in this cohort. On multivariate analysis, the volume of lung spared from doses > or = 5 Gy (VS5) was the only significant independent factor associated with postoperative pulmonary complications (p = 0.005). CONCLUSIONS: Dosimetric factors but not clinical factors were found to be strongly associated with the incidence of postoperative pulmonary complications in this cohort of esophageal cancer patients treated with concurrent chemoradiation plus surgery. The volume of the lung spared from doses of > or = 5 Gy was the only independent dosimetric factor in multivariate analysis. This suggests that ensuring an adequate volume of lung unexposed to radiation might reduce the incidence of postoperative pulmonary complications. PMID- 16242258 TI - Biologically effective dose values for prostate brachytherapy: effects on PSA failure and posttreatment biopsy results. AB - PURPOSE: To analyze the effect of biologically effective dose (BED) values on prostate-specific antigen (PSA) failure and posttreatment biopsy. METHODS AND MATERIALS: From 1990 to 2003, 1,377 patients had prostate brachytherapy alone (I 125 or Pd-103) (571), hormonal and brachytherapy (371), and trimodality therapy (hormonal, implant, and external beam) (435). Dose was defined as the D90 (dose delivered to 90% of the gland from the dose-volume histogram). RESULTS: Freedom from PSA failure (FFPF) at 10 years was 87%. The 10-year FFPF for BED<100, >100 120, >120-140, >140-160, <160-180, >180-200, and >200 were 46%, 68%, 81%, 85.5%, 90%, 90%, and 92%, respectively (p<0.0001). BED and Gleason score had the greatest effect, with p values of p<0.0001 in multivariate analysis. Posttreatment positive biopsy rate was 7% (31/446). The positive biopsy rates for BED100-120, >120-140, >140-160, >160-180, >180-200, and >200 were 24% (8/33), 15% (3/20), 6% (2/33), 6% (3/52), 7% (6/82), 1% (1/72), and 3% (4/131), respectively (p<0.0001). BED was the most significant predictor of biopsy outcome in multivariate analysis (p=0.006). CONCLUSIONS: Biologically effective dose equations provide a method of comparing different isotopes and combined therapies in the brachytherapy management of prostate cancer. The effects of BED on FFPF and posttreatment biopsy demonstrate a strong dose-response relationship. PMID- 16242259 TI - The radiation-induced changes in rectal mucosa: hyperfractionated vs. hypofractionated preoperative radiation for rectal cancer. AB - PURPOSE: The purpose of the study was the qualitative and quantitative evaluation of acute radiation-induced rectal changes in patients who underwent preoperative radiotherapy according to two different irradiation protocols. PATIENTS AND METHODS: Sixty-eight patients with rectal adenocarcinoma underwent preoperative radiotherapy; 44 and 24 patients underwent hyperfractionated and hypofractionated protocol, respectively. Fifteen patients treated with surgery alone served as a control group. Five basic histopathologic features (meganucleosis, inflammatory infiltrations, eosinophils, mucus secretion, and erosions) and two additional features (mitotic figures and architectural glandular abnormalities) of radiation induced changes were qualified and quantified. RESULTS: Acute radiation-induced reactions were found in 66 patients. The most common were eosinophilic and plasma cell inflammatory infiltrations (65 patients), erosions, and decreased mucus secretion (54 patients). Meganucleosis and mitotic figures were more common in patients who underwent hyperfractionated radiotherapy. The least common were the glandular architectural distortions, especially in patients treated with hypofractionated radiotherapy. Statistically significant differences in morphologic parameters studied between groups treated with different irradiation protocols were found. CONCLUSION: The system of assessment is a valuable tool in the evaluation of radiation-induced changes in the rectal mucosa. A greater intensity of regenerative changes was found in patients treated with hyperfractionated radiotherapy. PMID- 16242261 TI - Bio-guided isolation of the cytotoxic corytenchine and isocoreximine from roots of Annona cherimolia. AB - The roots of Annona cherimolia have been shown to contain the known tetrahydroprotoberberine alkaloids corytenchine (1) and isocoreximine (2), investigated for their cytotoxic properties. PMID- 16242260 TI - A phase I study of concurrent radiotherapy and capecitabine as adjuvant treatment for operable rectal cancer. AB - PURPOSE: To determine the maximum tolerated dose and the dose-limiting toxicity of capecitabine with standard radiotherapy (RT) as adjuvant treatment in patients with rectal cancer. METHODS AND MATERIALS: Patients with Stage II/III rectal cancer after surgery were eligible. Total RT dose was delivered as DT 50 Gy in fractions of 2.0 Gy/day for 5 weeks to the pelvic area. Capecitabine was administered concurrently with RT in escalating doses, twice daily with a 12-h interval, for two cycles of 14 days separated by a 7-day rest. Dose-limiting toxicity included Grade 3 or Grade 4 hematologic and nonhematologic toxicity. RESULTS: Twenty-four patients were enrolled at the following dose levels: 1,000 (3 patients), 1,200 (3 patients), 1,400 (3 patients), 1,500 (3 patients), 1,600 (6 patients), and 1,700 mg/m2/day (6 patients). Dose-limiting toxicity was observed in 1 patient at 1,600 mg/m2/day (Grade 3 diarrhea) and in 2 patients at 1,700 mg/m2/day (1 patient had Grade 3 and 1 Grade 4 diarrhea). CONCLUSION: The maximum tolerated dose (MTD) of capecitabine given concurrently with RT was 1,600 mg/m2, daily from the 1st to the 14th day, with a 7-day rest, for two cycles. PMID- 16242262 TI - A new kaurane diterpene dimer from Parinari campestris. AB - A new kaurane diterpene dimer, 15-oxozoapatlin-13alpha-yl-10'alpha,16'alpha dihydroxy-9'alpha-methyl-20'-nor-kauran-19'-oic acid gamma-lactone-17'-oate (1), together with the known 13-hydroxy-15-oxozoapatlin (2), 10alpha,13alpha,16alpha,17-tetrahydroxy-9alpha-methyl-15-oxo-20-nor-kauran-19-oic acid gamma-lactone (3), 2alpha,10alpha,13alpha,16alpha,17-pentahydroxy-9alpha methyl-15-oxo-20-nor-kauran-19-oic acid (19,10)-lactone (4), 3alpha,10alpha,13alpha,16alpha,17-pentahydroxy-9alpha-methyl-15-oxo-20-nor-kauran 19-oic acid gamma-lactone (5), and 1beta,16alpha,17-trihydroxy-ent-kaurane (6) were isolated from the leaves of Parinari campestris and identified on the basis of detailed spectral analysis, including 2D NMR spectrometry and ESI-MS. PMID- 16242263 TI - Compositions and antimicrobial activities of the essential oils of two Hypericum species from Turkey. AB - The chemical compositions of essential oils obtained from Hypericum hyssopifolium var. microcalycinum and Hypericum lysimachioides var. lysimachioides were analysed by using GC and GC-MS. Caryophyllene oxide was found to be the major component. The essential oils of both Hypericum species showed antimicrobial activity against nine microorganism at a concentration of 60 to 80 microg/ml. PMID- 16242264 TI - Screening of some rare endemic Italian plants for inhibitory activity on 5 lipoxygenase. AB - The extracts of four rare plants found on the islands of Sicily, Vulcano and Marettimo, Southern Italy, were screened for their inhibitory effect on the production of leukotriene B4 by 5-lipoxygenase in intact cells. The methanol extracts of pods of Cytisus aeolicus and aerial parts of Thymus richardii were the most active extracts, inhibiting almost completely the leukotriene B4 production at 200 and 50 microg/ml, respectively. PMID- 16242265 TI - Leaf and stem morphoanatomy of Petiveria alliacea. AB - Petiveria alliacea is a perennial herb native to the Amazonian region and used in traditional medicine for different purposes, such as diuretic, antispasmodic and anti-inflammatory. The morphoanatomical characterization of the leaf and stem was carried out, in order to contribute to the medicinal plant identification. The plant material was fixed, freehand sectioned and stained either with toluidine blue or astra blue and basic fuchsine. Microchemical tests were also applied. The leaf is simple, alternate and elliptic. The blade exhibits paracytic stomata on the abaxial side, non-glandular trichomes and dorsiventral mesophyll. The midrib is biconvex and the petiole is plain-convex, both traversed by collateral vascular bundles adjoined with sclerenchymatic caps. The stem, in incipient secondary growth, presents epidermis, angular collenchyma, starch sheath and collateral vascular organization. Several prisms of calcium oxalate are seen in the leaf and stem. PMID- 16242266 TI - Antibacterial and cytotoxic compounds from the bark of Cananga odorata. AB - O-Methylmoschatoline, liriodenine and 3,4-dihydroxybenzoic acid isolated from the barks of Cananga odorata showed antibacterial activities against a number of Gram (+) and Gram (-) bacteria. The compounds also showed antifungal and cytotoxic activities. PMID- 16242267 TI - A new benzophenanthridine alkaloid from Zanthoxylum buesgenii. AB - A new benzophenanthridine alkaloid, 6-[2'-ethoxy-2'-(2'',4'',5'' trimethoxyphenyl)] ethyl-7,8-dimethoxy-5-methyl-2,3-methylenedioxy-5,6 dihydrobenzo[c]phenanthridine named buesgeniine (1), as well as the known decarine, were isolated from the extract of the stem bark of Zanthoxylum buesgenii. In addition, three known lignans, sesamine, matairesinol dimethylether, and methylpluviatilol, were also identified. The structure of 1 was elucidated using spectroscopic methods. PMID- 16242268 TI - Antimicrobial activity of 10-isobutyryloxy-8,9-epoxythymol isobutyrate. AB - 10-Isobutyryloxy-8,9-epoxythymol isobutyrate was found to be a major constituent of Inula helenium and Inula royleana root cultures. The compound showed moderate antimicrobial activity against Staphylococcus aureus, Enterococcus faecalis, Escherischia coli, Pseudomonas aeruginosa and Candida albicans. PMID- 16242269 TI - Asterosaponins from the starfish Culcita novaeguineae and their bioactivities. AB - Bioassay-guided fractionation of the n-BuOH extract of the starfish Culcita novaeguineae resulted in the isolation of one new sulfated steroidal glycoside (asterosaponin) (1), along with three known asterosaponins, thornasteroside A (2), marthasteroside A(1) (3) and regularoside A (4), as active compounds causing morphological abnormality of Pyricularia oryzae mycelia. Their structures were elucidated by extensive spectral studies and chemical evidences. All the saponins showed moderate cytotoxicity against cancer cell lines K-562 and BEL-7402. PMID- 16242270 TI - Anti-nociceptive activity of the aqueous extract of Erythrina velutina leaves. AB - Anti-nociceptive and anti-oedematogenic effects of the aqueous extract from the leaves of Erythrina velutina were tested through experimental models of nociception in mice and paw oedema induced by carrageenin in rats. The extract (300 and 600 mg/kg) did not change the carrageenin-induced paw oedema. In the hot plate test the extract also did not alter the latency time for mice liking the rear paws. Moreover, the extract (600 mg/kg) decreased by 96.5% the paw liking time in the second phase of the formalin test. This effect was antagonized by naloxone (5 mg/kg). In the acetic acid-induced writhing test, the extract (300 and 600 mg/kg) reduced the number of writhing by 88.8% and 96.4%, respectively. Our present results demonstrated that the crude extract from the leaves of E. velutina has anti-nociceptive but not anti-oedematogenic properties. PMID- 16242271 TI - Insect antifeedant and growth regulating activities of salannobutyrolactone and desacetylsalannobutyrolactone. AB - Salannobutyrolactone and its deacetylderivative were tested for insect antifeedant and growth regulatory activities against the tobacco cutworm, Spodoptera litura. Salannobutyrolactone was the most effective antifeedant. Desacetylsalannobutyrolactone increased larval duration and larval mortality. PMID- 16242272 TI - Housekeeping recA gene interrupted by group II intron in the thermophilic Geobacillus kaustophilus. AB - Most of group II introns are found in intergenes and CDSs with unknown functions, but not in housekeeping genes. In particular, no group II intron within the housekeeping recA gene has been reported either in eukaryotic genomes or in prokaryotic genomes. In this study, we found that the recA gene of the thermophilic Geobacillus kaustophilus genome is interrupted by a group II intron (Gk. Int1), and that Gk.Int1 can splice in temperatures above 70 degrees C in vivo. Here, we report the first prokaryotic group II intron to be found in a housekeeping gene, the characteristics of its self-splicing in vivo and in vitro, and our conclusion that the recA gene functions through the self-splicing of Gk.Int1. It is suggested that the amelioration of Gk.Int1 intron has occurred recently, and that it is still in the process of evolution to the recipient genome. PMID- 16242273 TI - Recombinant bovine interleukin 2 enhances immunity and protection induced by Brucella abortus vaccines in cattle. AB - Augmentation of immunization of cattle Brucella abortus S19 or a B. abortus soluble protein extract (SPEBA) vaccine through administration of recombinant bovine IL 2 (rBoIL 2) was evaluated. Seventy-five heifers were divided among 6 groups that were treated with the following: Group 1, no treatment; Group 2, rBoIL 2 (1microg/kg) on day 0; Group 3, SPEBA (2 mg) on day 0 and week 9; Group 4, SPEBA + rBoIL 2 on day 0, SPEBA on week 9; Group 5, S19 (10(7) CFU) on day 0 and week 9; Group 6, S19 + rBoIL 2 on day 0, S19 only on week 9. Approximately, 6 months after vaccination, cattle were bred by natural service, and at mid gestation pregnant cattle were challenged intraconjunctivally with 9.1 x 10(5) CFU of virulent B. abortus S2308. Pre- and post-challenge antibody responses were measured by an enzyme-linked immunosorbent assay, a particle concentration fluorescence assay, and the card test. Lymphoproliferation (LP) responses to gamma-irradiated B. abortus and SPEBA antigens were measured in peripheral blood mononuclear cells. After vaccination, antibody responses to B. abortus elevated rapidly in SPEBA- and S19-vaccinates with and without rBoIL 2, however, these responses were significantly (P < 0.05) higher in vaccinates which also received rBoIL 2. Antibody levels for all vaccinated groups had returned to those of negative control groups by the challenge date with the exception of the SPEBA/rBoIL 2 group. In general, LP responses were higher in vaccinated or rBoIL 2-treated cattle than for unvaccinated controls. Challenge of 48 pregnant heifers resulted in abortions in 4/9 of Group 1, 0/9 of Group 2, 4/8 of Group 3, 2/9 of Group 4, 1/7 of Group 5, and 0/6 of Group 6 cattle. Treatment with rBoIL 2 alone (Group 2) provided significant (P < 0.05) protection from infection, abortions and induction of sero-positive status compared to untreated (Group 1) cattle. Co administration of rBoIL 2 with S19 resulted in significant (P < 0.05) augmentation in onset, duration and magnitude of LP responses to B. abortus antigens following challenge. Characterization of the cytokine response of bovine monocyte-derived macrophages by real-time polymerase chain reaction indicated that in vitro stimulation of these cells with rBoIL 2 resulted in a profound up regulation of genes encoding tumor necrosis factor-alpha, IL 12p40, and interferon-gamma reflecting activation of the cells. Overall, rBoIL 2-treatment was associated with fewer infections, sero-conversions and a significant (P = 0.02) level of protection against abortion as compared to vaccination alone or no treatment. PMID- 16242274 TI - Identification of mammalian cell genotoxins in respirable diesel exhaust particles by bioassay-directed chemical analysis. AB - A bioassay-directed chemical analysis which consists of mammalian cell bioassays (comet assay, CBMN assay and EROD-microbioassay) in conjunction with analytical measurements was performed to identify the most biologically active compounds of the diesel exhaust particulate matters (DEPs) on mutagenic activity. These bioassay systems were suitable to estimate the mammalian genotoxic potentials of pollutants present in low concentrations in limited environmental samples, as is the case with DEPEs. The results from mutagenic assay showed that the aromatic and slightly polar fraction of DEPs induced chromosomal damage and DNA breakage in a non-cytotoxic dose. It was also revealed that indirect-acting mutagens may mainly contribute to the mutagenic effect of aromatic fraction via the enzyme metabolism system. In the aromatic fraction, several indirect-acting mutagenic PAHs such as dibenzo(a,h)anthracene, chrysene, and 1,2-benzanthracene were detected by GC-MS and the complex mixture effect of this fraction was quantified in terms of its biological-TCDD equivalent concentration (bio-TEQ) which was 32.82 bio-TEQ ng/g-DEPs by EROD-microbioassay. Conclusively, we confirmed that indirect-acting mutagens contained in aromatic fraction may be important causatives of the genotoxicity of extracts of DEPs by integrating the results obtained from a mammalian cell bioassay-directed fractionation. PMID- 16242275 TI - Reference values of coplanar and non-coplanar PCBs in serum samples from two Italian population groups. AB - The main goal of this study is to establish the reference values of individual Polychlorinated biphenyl (PCB) congeners in non-occupationally exposed subjects. Since the PCB pattern in human serum is related to the living area, two different population groups from North and Central Italy, were compared. Serum concentrations of both coplanar and non-coplanar PCB congeners were measured by using gas chromatography coupled with low-resolution mass spectrometry (HRGC LRMS). A fast and reliable method for the determination of 60 congeners had been previously validated. Its reliability was further verified by using high resolution mass spectrometry. Thirty-one congeners out of 60 were found at detectable concentrations in at least one sample. The mean value for total PCBs was found to be 2.48 and 3.93 microg/L for the two population groups. Eight dioxin-like PCBs were detected. In accordance with the findings from the literature, the most abundant congeners were found to be 153, 138, 180, and 170. Both univariate and multivariate analysis showed that age is a significant determinant of PCB concentrations. The correlation increased with increasing chlorination. Slight differences in the PCB pattern were observed in the two population groups. PMID- 16242277 TI - Spray-freeze-dried liposomal ciprofloxacin powder for inhaled aerosol drug delivery. AB - Spray-freeze drying was utilized to manufacture a liposomal powder formulation containing ciprofloxacin as a model active component. The powder forms liposomally encapsulated ciprofloxacin when wetted. Aerosol properties of this formulation were assessed using a new passive inhaler, in which the powder was entrained at a flow rate of 60l/min. A mass median aerodynamic diameter (MMAD) of 2.8 microm was achieved for this formulation. Using the experimental dispersion testing data, ciprofloxacin concentration in the airway surface liquid (ASL) was calculated using a Lagrangian deposition model. The reconstitution of the powder in various aqueous media gave drug encapsulation efficiencies as follows: 50% in water, 93.5% in isotonic saline, 80% in bovine mucin, 75% in porcine mucus and 73% in five-fold-diluted ex vivo human cystic fibrosis patient sputum. PMID- 16242276 TI - Drug release and permeation studies of nanosuspensions based on solidified reverse micellar solutions (SRMS). AB - Solidified reverse micellar solutions (SRMS), i.e. mixtures of lecithin and triglycerides, offer high solubilisation capacities for different types of drugs in contrast to simple triglyceride systems [Friedrich, I., Muller-Goymann, C.C., 2003. Characterisation of SRMS and production development of SRMS-based nanosuspensions. Eur. J. Pharm. Biopharm. 56, 111-119]. Nanosuspensions based on SRMS were prepared by homogenisation close to the melting point of the SRMS matrix. In a first step the SRMS matrices of 1:1 (w/w) ratios of lecithin and triglycerides were loaded with 17beta-estradiol-hemihydrate (EST), hydrocortisone (HC) or pilocarpine base (PB), respectively, and subsequently ground in liquid nitrogen to minimise drug diffusion later on. The powder was then dispersed in a polysorbate 80 solution using high pressure homogenisation. The drug loading capacities of the nanosuspensions were very high in the case of poorly water soluble EST (99% of total 0.1%, w/w, EST) and HC (97% of total 0.5%, w/w, HC) but not sufficient with the more hydrophilic PB (37-40% of total 1.0%, w/w, PB). These findings suggest SRMS-based nanosuspensions to be promising aqueous drug carrier systems for poorly soluble drugs like EST and HC. Furthermore, in vitro drug permeation from the different drug-loaded nanosuspensions was performed across human cornea construct (HCC) as an organotypical cell culture model. PB permeation did not differ from the nanosuspension and an aqueous solution whereas the permeation coefficients of HC-loaded nanosuspensions were reduced in comparison to aqueous and oily solutions of HC. However, the permeated amount was higher from the nanosuspensions due to a much lower HC concentration in the solution than that in the nanosuspension (solution 0.02%, w/w, versus nanosuspension 0.5%, w/w). The high drug load of the nanoparticles provides prolonged HC release. Permeated amounts of EST were reduced in comparison to HC and only detectable with an ELISA technique. The EST release from nanosuspensions and different EST-loaded systems revealed a prolonged EST release from the nanoparticulate systems in contrast to a faster release of an oily solution of an equal EST concentration. With regard to an aqueous EST suspension of similar concentration which represents a depot system the release rate from the nanosuspensions revealed the same order of magnitude which points again to a prolonged release potential of the nanosuspensions. PMID- 16242279 TI - Effects of Urginea sanguinea, a traditional asthma remedy, on embryo neuronal development. AB - The Southern African plant, Urginea sanguinea Shinz (Hyacinthaceae) (US), is a well-known traditional herbal medicine and it is used for many different ailments, including asthma. Pregnant women also use this plant and little is known regarding the toxic effects of this plant material on the developing foetus. US contains the cardiac glycoside (CG) Transvaalin; CGs are known to cross the placenta and blood-brain barrier and therefore may have a negative effect on the foetal development. To address this, in vitro cytotoxicity of this preparation as well as its effect on chick embryo neural development was investigated. Water extracts of US were shown to be cytotoxic in cell cultures of L929 cell and primary embryonic neural cell cultures. Electron microscopy studies following in ovo exposure revealed altered neuron morphology with patterns of cell damage either associated with apoptosis or necrosis. CGs are known to inhibit membrane bound Na(+)/K(+)-ATPase in conducting tissues, causing disruption of the calcium pathways, mitochondrial calcium overload leading to either apoptosis or necrosis or where both occur, a process of necrapoptosis. The in ovo effects observed strongly indicate that US causes necrapoptosis in chick embryonic neurons. PMID- 16242278 TI - Synthesis, physicochemical properties and antiviral activities of ester prodrugs of ganciclovir. AB - The purpose of this study was to synthesize a series of diester prodrugs of ganciclovir (GCV), for improving ocular and oral bioavailability and therapeutic activity. Solubility, logP, pH stability profile, in vitro antiviral activity, cytotoxicity, inhibition profile and ocular tissue hydrolysis of the GCV prodrugs were measured. Val-Val-GCV and Val-Gly-GCV diesters were found to exhibit greater aqueous stability compared to Val-GCV and Gly-Val-GCV while ocular tissue hydrolysis demonstrated Val-Gly-GCV and Gly-Val-GCV to be more stable. Val-Val GCV and Val-GCV diesters were the most lipophilic compounds and were predicted to possess a partition coefficient 295- and 12-fold greater than that of GCV, respectively. All the prodrugs possess much higher aqueous solubility than the parent drug GCV. Ex vivo uptake in the rabbit eye indicates that the prodrugs have high uptake potential. The prodrugs showed no increase in cytotoxicity compared to GCV, instead there was a marked increase in their potency against human cytomegalovirus (HCMV) as well as HSV-1 and HSV-2. This should allow therapeutic response to be seen at a lower concentration that can be achieved more easily, than the drugs currently being used. In conclusion, the diester GCV prodrugs demonstrated excellent chemical stability, high aqueous solubility and markedly enhanced antiviral potency against the herpes viruses without any increase in cytotoxicity. PMID- 16242280 TI - Liquid chromatographic assay of ivermectin in human plasma for application to clinical pharmacokinetic studies. AB - There is a need for an accurate, sensitive and selective high-performance liquid chromatography (HPLC) method for the quantitation of ivermectin in human plasma that separates the parent drug from metabolites. Ivermectin and the internal standard, moxidectin, were extracted from 0.2 ml of human plasma using Oasis HLB solid phase extraction cartridges. After extraction, fluorescent derivatives of ivermectin and moxidectin were made by reaction with trifluoroacetic anhydride and N-methylimidazole. Separation was achieved on a Alltech Ultrasphere C18 5mu column with a mobile phase composed of tetrahydrofuran-acetonitrile-water (40:38:22 v/v/v). Detection is by fluorescence, with an excitation of 365 nm and emission of 475 nm. The retention times of ivermectin and internal standard, moxidectin are approximately 24.5 and 12.5 min, respectively. The assay is linear over the concentration range of 0.2-200 ng/ml of ivermectin in human plasma (r = 0.9992, weighted by 1/concentration). Recoveries of ivermectin are greater than 80% at all concentrations. The analysis of quality control samples for ivermectin 0.2, 25, and 200 ng/ml demonstrated excellent precision with coefficient of variation of 6.1, 3.6 and 2.3%, respectively (n = 6). The method is accurate with all intra-day (n = 6) and interday (n = 12) mean concentration within 10% of nominal values at all quality control sample concentrations. Storage stability for 30 days at -80 degrees C and after three freeze-thaw cycles are within acceptable limits. The method separates ivermectin from multiple less and more polar unidentified metabolites. This method is robust and suitable for clinical pharmacokinetic studies. The analytical procedure has been applied to a pharmacokinetic study of ivermectin in healthy volunteers and to the analysis of plasma specimens from patients with disseminated strongyloidiasis. PMID- 16242281 TI - Multiplexed cytokine detection in microliter microdialysis samples obtained from activated cultured macrophages. AB - Microdialysis sampling probes were used to collect cytokine samples from lipopolysaccharide (LPS)-stimulated macrophages. The probes were immersed into cell culture wells containing either RAW 264.7 or isolated peritoneal macrophages. Dialysates (15 microL) from these wells were subjected to a multiplexed cytokine sandwich immunoassay platform analyzed by flow cytometry that measures up to six separate cytokines, interleukin-6 (IL-6), interleukin-10 (IL-10), interleukin-12p70 (IL-12p70), interferon-gamma (IFN-gamma), macrophage chemoattractant protein-1 (MCP-1), and tumor necrosis factor-alpha (TNF-alpha) in a single 15-muL sample. In vitro microdialysis sampling relative recovery experiments showed that only IFN-gamma, IL-6, MCP-1, and TNF-alpha could be recovered across a commercially-available 100-kDa MWCO microdialysis membrane. Eleven hours after LPS addition (1 microg/mL), RAW 264.7 macrophages secreted greater than 8000 pg/mL of TNF-alpha and greater than 1000 pg/mL MCP-1. With the peritoneal macrophages, greater than 6000 pg/mL of IL-6, MCP-1, and TNF-alpha were obtained. The maximum dialysate concentrations obtained from the RAW macrophages were 1300 pg/mL for TNF-alpha and 55 pg/mL for MCP-1. Maximum cytokine concentrations from peritoneal macrophage dialysates reached approximately 2000 pg/mL, 1100 pg/mL and 500 pg/mL for TNF-alpha, MCP-1 and IL-6, respectively. Microdialysis sampling allowed for 20-min samples to be collected during the cytokine release from the activated macrophages. These results demonstrate that microdialysis sampling can be used for collection of selected cytokines with improved temporal resolution. PMID- 16242282 TI - Determination of bovine serum albumin by a resonance light-scattering technique with the mixed-complex La(Phth)(phen)(3+). AB - A mixed-complex of La(Phth)(phen)(3+) was synthesized. Resonance light-scattering characteristics of interaction between La(Phth)(phen)(3+) with bovine serum albumin (BSA) were studied. When BSA was added, aggregation of La(Phth)(phen)(3+) on the molecular surface of bovine serum albumin occurred in the pH 5.5-6.3, resulted in an enhanced resonance light-scattering (RLS) peak at 360 nm. The intensity of resonance light-scattering was found to be proportional to the concentration of BSA. PMID- 16242283 TI - Enhancement of melatonin photostability by encapsulation in lipospheres. AB - The effect of lipid microparticle carrier systems on the light-induced degradation of melatonin was investigated. Microspheres loaded with melatonin were prepared using tristearin or tripalmitin as the lipid material and hydrogenated phosphatidylcholine or polysorbate 60 as the emulsifier. The obtained lipid microspheres were characterized by scanning-electron microscopy and differential scanning calorimetry. Free or microencapsulated melatonin was incorporated in a model cream formulation (oil-in-water emulsion) and irradiated with a solar simulator. The extent of photodegradation was measured by high performance liquid chromatography. The photolysis experiments demonstrated that the light-induced decomposition of melatonin was markedly decreased by encapsulation into lipid microspheres based on tristearin and phosphatidylcholine (the extent of degradation was 19.6% for unencapsulated melatonin compared to 5.6% for the melatonin-loaded microparticles). Therefore, incorporation in lipid microparticles can be considered an effective system to enhance the photostability of melatonin. PMID- 16242284 TI - Interference from a glucuronide metabolite in the determination of ramipril and ramiprilat in human plasma and urine by gas chromatography-mass spectrometry. AB - In the course of development and validation of a gas chromatography-mass spectrometry (GC-MS) method for ramipril and its biologically active metabolite ramiprilat, evidence was found for an unknown interfering metabolite. Sample treatment included isolation from plasma or urine by solid-phase extraction, methylation with trimethylsilyldiazomethane and acylation with trifluoroacetic anhydride (TFAA). When liquid chromatography was used to fractionate plasma extracts prior to derivatization, the alkyl, acyl-derivative of ramipril was obtained from two separate LC fractions. Electrospray ionization mass spectral data, together with circumstances for the derivatization, were consistent with the presence of an N-glucuronide of ramipril. Interference from the metabolite was eliminated by including a wash step after extraction/alkylation, prior to acylation. The final assay had a lower limit of quantification at 1.0 nmol/L and a linear range of 1-300 nmol/L. Intra- and inter-batch precision for ramipril and ramiprilat in plasma or urine were better than 10 and 5% at 2 and 80 nmol/L, respectively. PMID- 16242285 TI - Novel stir bar sorptive extraction methods for environmental and biomedical analysis. AB - Stir bar sorptive extraction (SBSE) is sample preparation technique that involves the extraction and enrichment of organic compounds from a liquid sample. The technique is based on the principle of sorptive extraction. A large amount of extraction phase is coated on a stir bar. An analyte is extracted into the extraction phase, based on its octanol-water partitioning coefficient and the phase ratio. Recently, various methods involving SBSE were developed in order to further facilitate analysis and improve sensitivity. In this review, we focused on the novel methods that involve SBSE with in situ derivatization, SBSE with in situ de-conjugation, thermal desorption (TD) in the multi-shot mode and TD with in tube derivatization method. Those methods were applied successfully to the trace analysis of environmental and biological samples and extremely low detection limits were achieved. PMID- 16242286 TI - A reversed phase HPLC assay for the simultaneous quantitation of non-ionic and ionic surfactants in bioprocess intermediates. AB - This report describes a rapid and accurate reversed phase HPLC method for the simultaneous quantitation of multiple surfactants in various bioprocess solution matrices including cell lysates. Separation and quantitation of a mixture of the cationic detergent domiphen bromide from the non-ionic detergent Triton X-100 in crude cell mixtures can be achieved within 15 min using a TSK-gel C18-NPR reversed phase column and an aqueous mobile phase gradient of acetonitrile:water with the reagent PIC-B8 as ion-pairing modifier. The linear dynamic range for quantitation of domiphen bromide (DB) and Triton in this assay extends from 20 to 2000 microM. Linear regression analyses from the standard curve determinations showed an R2 of > or = 0.990. The assay does not show any interferences from proteins or other cellular contaminants such as nucleic acids. The assay has been used to evaluate clearance of these compounds throughout the purification process of an adenovirus-based vaccine candidate, as well as to determine the effects of process changes on detergent clearance. PMID- 16242287 TI - Simultaneous determination of active ingredients in an ophthalmic solution by isocratic tandem-mode HPLC connected reverse phase column and strong cation exchange column. AB - A tandem-mode high performance liquid chromatography (HPLC) system is described here, which employs reversed phase liquid chromatography (RPLC) followed by strong cation exchange liquid chromatography (SCX), was used to determine the mixture of six ingredients in an ophthalmic solution. As a result of investigations, isocratic HPLC methods that using two columns in tandem-mode; Atlantis dC18 (75 mm x 4.6 mm i.d., 3 microm, ODS) and CAPCELL PAK SCX UG80 (75 mm x 4.6 mm i.d., SCX), which have different separation modes, and control of mixture of methanol/ammonium dihydrogenphosphate buffer as used for the eluent, allowed for six target ingredients to be determined simultaneously. And all ingredients separated perfectly and were determined efficiently and rapidly. Validation of the method was accomplished with respect to linearity (r>0.999), recovery (99.4-100.4%), precision (R.S.D. 0.1-0.9%) and specificity. These results suggest that the fusion of different separation modes can be used for the simultaneous determination of ingredients in ophthalmic solutions, and this can be accomplished rapidly and with high precision. PMID- 16242288 TI - Improved LC of minocycline drug substance. AB - An isocratic liquid chromatographic method is described for the separation of minocycline and its impurities. This method uses XTerra RP-18, 5 microm (25 cm x 4.6 mm I.D.), a silica-based stationary phase with reduced silanol activity. A mobile phase composed of acetonitrile-0.2 M tetrabutylammonium hydrogen sulphate pH 6.5-0.2 M ethylenediaminetetraacetic acid pH 6.5-water (20:20:20:40; v/v/v/v) was used at a flow rate of 1 ml/min. The column temperature was set at 35 degrees C. UV detection was performed at 280 nm. Optimisation of the separation method and a robustness study were performed by means of a central composite experimental design. The method allows to separate minocycline from known impurities. Some unidentified impurities are also separated. The total time of analysis is less than 20 min. PMID- 16242289 TI - Sequential-injection determination of traces of disodium phenyl dibenzimidazole tetrasulphonate in urine from users of sunscreens by on-line solid-phase extraction coupled with a fluorimetric detector. AB - A sensitive and selective method to determine disodium phenyl dibenzimidazole tetrasulphonate (PDT) in the urine of sunscreen users, which is suitable for studies on body accumulation/excretion is proposed. On-line solid-phase extraction allows the analyte to be retained and subsequentely eluted, using a strong anion exchange (SAX) microcolumn. Standard addition calibration was carried out with only one standard. The wavelengths of excitation and emission were 330 and 454 nm, respectively. The method allows PDT to be determined in both, spiked and unspiked human urine samples, without any pre-treatment. Results obtained for spiked urine samples (40-200 ng ml(-1)) showed the accuracy of the method. The mean relative standard deviations (R.S.D.) of the results was 7%. Five volunteers applied a sunscreen lotion containing 5% PDT and their urinary excretion was controlled from the moment of application until the excreted amounts were no longer detectable. The sensitivity of the proposed method is in the order of 1900 ml microg(-1) and the detection limit (3S(y/x)/b) is in the order of 5 ng of PDT, which means 10 ng ml(-1) for a 500 microl injected volume, and this is suitable for the PDT levels found in the urine. PMID- 16242290 TI - Determination of bifendate in human plasma by HPLC-MS and bioequivalence on bifendate pills in healthy volunteers. AB - A sensitive and specific method for the determination of bifendate in human plasma was developed, based on high-performance liquid chromatography (HPLC)-mass spectrometry (MS). The samples were extracted from plasma with diethyl ether, followed by separation and evaporation after addition of internal standard diazepam. The residue was reconstituted in methanol and injected into the HPLC MS. Chromatography was performed on an Inertsil ODS column with a mobile phase consisting of methanol-distilled water (70/30, v/v) at a flow rate of 0.3 mL/min. Quantitative analysis was achieved by MS detection, using a mass spectrometer equipped with an electrospray ionization interface (ESI) and operated in selected ion monitoring (SIM) and positive-ionization mode using target ions at m/z 419 for bifendate and m/z 285 for internal standard, respectively. The linearity was confirmed in the concentration range of 2-200 ng/mL in human plasma and the precision of this assay was not more than 6.79% over the entire concentration range. The method was sensitive and repeatable enough to be used in pharmacokinetic and bioavailability studies. PMID- 16242291 TI - Enhancing parent-provider communication in ambulatory pediatric practice. AB - OBJECTIVE: To address the need for interventions to enhance parent-provider communication, the present study evaluated the effectiveness of a brief intervention to enhance interpersonal communication skills by pediatric and internal medicine/pediatric residents with parents of children seen in ambulatory practice. METHODS: In a within-subjects, repeated measures research design, residents (N=28) participated in a 1-112h intervention designed to increase the use of effective communication skills with their patients' families (N=92). Three pediatric visits (one prior to and two following the intervention) were audio taped and coded using Roter's Interaction Analysis System. Parents' perceptions of residents' communication skills and satisfaction with care were gathered. RESULTS: Residents used more effective interpersonal communication skills following the intervention (p<.05). Parent satisfaction with care also increased (p<.01) with parents reporting greater satisfaction with distress relief (p=.01) and communication following the intervention (p=.05). However, parents' perceptions of resident communication did not change over time. CONCLUSION: These findings suggest that a relatively brief intervention shows promise in improving the quality of care provided within a pediatric ambulatory care setting. PRACTICE IMPLICATIONS: Training residents to increase their use of praise, empathy, and collaboration with families can be done within a brief intervention and may be associated with increased parental satisfaction with care. PMID- 16242292 TI - Patients' perceptions of visit companions' helpfulness during Japanese geriatric medical visits. AB - OBJECTIVE: Elderly patients are often accompanied during medical visits by a companion, usually a family member. This study explores the association between patients' expectations regarding the communication role of their companions, and the role intended and taken by companions during the medical visits, on patient perceptions of companion helpfulness. METHODS: Participants included 63 patients aged 65 or over who were under continuous care of nine attending physicians at a university-affiliated geriatric clinic in Tokyo, and their companions during the medical visit. These medical visits were audiotape recorded and coded using a Japanese translation of the Roter Interaction Analysis System (RIAS). RESULTS: The discrepancy between what patients expected of their companions, and what companions intended and actually did during the visit, predicted patient ratings of companion helpfulness. It was especially noteworthy that the highest ratings of companion helpfulness occurred when patients expected their companion to be actively involved in communication and they were. However, if the patient anticipated only a limited companion role, the companion was not viewed as very helpful regardless of the role they played. DISCUSSION: There would be a largely unexploited potential for improving communication effectiveness through the optimal engagement of companions in visit communication. PMID- 16242293 TI - A literature review of the psychological impact of genetic testing on breast cancer patients. AB - OBJECTIVE: Easier access and increased awareness results in more referral for genetic testing for hereditary breast cancer in healthy at-risk women and breast cancer patients. To investigate the psychological impact of genetic testing on breast cancer patients, literature pertaining to this group was reviewed. METHOD: Medline and PsychInfo databases were searched over the period 1995-2004 for studies aimed at breast cancer patients referred for genetic testing. Qualitative and quantitative psychological outcome measures were identified. RESULTS: Eight papers were identified focusing on women affected by breast cancer and undergoing genetic counseling and DNA testing. CONCLUSION: Genetic testing does not lead to an increase in psychological distress in breast cancer patients. However, a recent breast cancer diagnosis adds to general and cancer-specific distress prior to genetic counseling and after DNA test disclosure. PRACTICE IMPLICATIONS: Clinicians need to be aware of possible high psychological distress and additional counseling needs of recently diagnosed breast cancer patients taking part in genetic testing. Further research should focus on patients who decline genetic counseling or receive an inconclusive test result, including age upon and time since diagnosis. PMID- 16242294 TI - Infertility: a label of choice in the case of sexually dysfunctional couples. AB - OBJECTIVES: In Indian society infertility and sexual dysfunction are stigmatized characteristics. The aim of the study was to explore which label couples who have both complaints, prefer to describe their situation, and whether gender differ occur in this preference. METHODS: A total of 40 couples - all clients of the reproductive health clinic of the King Edward Memorial Hospital in Pune because of problems with infertility and sexual dysfunction - participated in this study. Data were collected by means of interviews. RESULTS: The findings show that sexual activity decreased as the number of childless years increased. However, the interspouse-relationship gets stronger and more supportive. The couples never revealed their sexual dysfunction to others. When the husband was sexually dysfunctional, the couples preferred to label their situation as 'infertility' in order to avoid stigma. CONCLUSION: Issues that are related to sexual dysfunction, sexual behavior, inter-spouse relationship and communication are important aspects into the reality of an infertile couple's life. PRACTICE IMPLICATION: In planning the treatment clinicians should give more attention to the specific cultural context of these aspects. PMID- 16242295 TI - Does stimulating self-care increase self-care behaviour for minor illnesses of Dutch and Turkish inhabitants of a deprived area in The Netherlands? AB - OBJECTIVE: The aim of the present study was to examine whether self-care behaviour increases after a self-care stimulating intervention that proved to be successful in reducing care-seeking behaviour for minor illnesses of Turkish and Dutch inhabitants of a deprived area in the Netherlands, and to see whether there are cultural differences. METHOD: This longitudinal study was based on a "pre test/post-test one group" design. Data were collected during three structured face-to-face interviews: before the intervention, and 6 months and 1 year after the intervention, in which GPs personally handed out booklets to their patients containing guidelines on the management of 12 minor illnesses. RESULTS: The number of self-reported self-care actions did not increase. In contrast to the Dutch, the Turkish participants reported a decrease in the number of self-care actions, their attitude towards self-care became more negative, and they perceived less control. CONCLUSION: Apparently, a reduction in formal health care utilisation is not engendered by an increase in self-care behaviour. In order to make sure that interventions like these will have the intended effect, more research is needed, particularly among non-western populations. PRACTICE IMPLICATIONS: In developing future healthcare-reducing interventions, one should be aware of possible unwanted side effects in non-western populations. PMID- 16242297 TI - Adherence is associated with the quality of professional-patient interaction in Directly Observed Treatment Short-course, DOTS. AB - OBJECTIVE: To investigate the association between the behaviour of health professionals as reported by patients, the quality of communication, patients' communication about their disease, and non-adherence to Directly Observed Tuberculosis Treatment Short-course, DOTS. METHODS: This study was designed as a case-control study based on 50 cases (non-adherents) and 100 controls (adherents), conducted in a hilly western district in Nepal. The participation rate was 80% for 50 cases and 95% for 100 controls. All covariates with p valueserine variation in the fifth extracellular domain. Allelic, haplotypic and genotypic frequencies did not differ significantly between the patient (n=134) and the control (n=227) population samples. Moreover, there is no evidence of interaction between the DSG1 and the HLA-DRB1 and IL6 genes, whose alleles had been found associated with differential susceptibility to PF. The results of this study agree with the described and predicted B- and T-cell epitopes of the dsg1 molecule, which seemingly are not affected by the allelic variation. We conclude that genetic diversity of the autoantigen dsg1 is not a major factor for PF pathogenesis in the Brazilian population. PMID- 16242305 TI - Genetic prediction of autoimmunity: initial oligogenic prediction of anti-islet autoimmunity amongst DR3/DR4-DQ8 relatives of patients with type 1A diabetes. AB - In this study, the combined risk for expressing anti-islet autoantibodies and type 1A diabetes (T1D) was prospectively examined in 85 sampled relatives who had the high-risk HLA genotype (DR3-DQ8 DR4-DQ2). An insulin gene polymorphism, -23 HphI, and a lymphocyte tyrosine phosphatase gene polymorphism at position 1858C>T (amino acid 620 Arg to Trp), PTPN22/LYP, were analyzed. Life tables were created evaluating time to anti-islet autoantibody development and T1D. Of relatives with the high-risk HLA type followed for 3years, 9 of 43 (28.1%) with the high-risk 23 HphI polymorphism developed anti-islet autoantibodies versus two of 36 (5.6%) relatives with the lower-risk -23 HphI genotypes (p=0.048). Of relatives with the high-risk HLA type followed for 5years, eight of 32 (25.0%) with the high-risk 23 HphI polymorphism (A/A) developed T1D versus zero of 26 (0%) relatives with the lower-risk -23 HphI genotypes (A/T and T/T) (p=0.006). The PTPN22/LYP polymorphism, with genotypes C/C, C/T, and T/T, did not show a significant difference in risk by genotype. These results highlight the multiplicative risk of combined high-risk genotypes at different loci in terms of time to autoantibody and autoimmune disease development. PMID- 16242306 TI - Apoptotic cells, autoantibodies, and the role of HMGB1 in the subcellular localization of an autoantigen. AB - A current model for the evolution of systemic lupus erythematosus hypothesizes that there is a genetic predisposition coupled with an environmental or infectious trigger. This study investigated whether apoptotic cells given with a proinflammatory signal could induce features of lupus. Balb/c mice were injected with an apoptotic Balb/c-derived myeloid cell line, J774.1, either with or without the DNA-binding protein HMGB1 for five injections over 16 days in an IACUC approved study. Mice were sacrificed at 6 weeks and 12 weeks after treatment. Renal disease was assessed by immunofluorescence and autoantibodies were defined by ELISA. Western blotting was performed to characterize autoantigens. Mice injected with apoptotic cells developed antibodies to histones, SSA, ssDNA, and phospholipids. Antibodies to SSA and ssDNA persisted; however, antibodies to histones, and phospholipid declined at 12 weeks. IgG deposits in the kidney were detected at 6 weeks and persisted through 12 weeks primarily in animals that received both apoptotic cells and HMGB1. Autoantibodies in mice were diverse but the mice that received apoptotic cells developed particularly high titer antibodies to an unknown 78kDa protein. This protein became externalized on the surface of J774.1 cells in the presence of HMGB1. Mice that received apoptotic J774.1 cells with HMGB1 developed more extensive renal IgG deposition. While the mechanism is uncertain, an important effect of HMGB1 was to alter the subcellular distribution of a major autoantigen, making the autoantigen accessible for immune responses. This is the first description of an inflammatory stimulus altering the immunologic availability of a potential autoantigen. PMID- 16242308 TI - Muscular activation patterns of healthy persons and low back pain patients performing a functional capacity evaluation test. AB - The results of most reported studies show differences between the muscular activity of low back pain patients and healthy subjects, but the focus has usually been on trunk muscles only, and they have not involved work-related tests or exercises. The reintegration of chronic low back pain patients to job market is a common problem. Therefore assessment systems like the functional capacity evaluation (FCE) according to Isernhagen [S.J. Isernhagen, Work Injury: Management and Prevention, Aspen Publishers Inc., Gaithersburg, MD, 1988] are often used tools to determine the physical abilities and deficits of long-time incapacitated persons. The aim of the present study was to compare the healthy persons and chronic low back pain patients in performing a FCE-test and to analyse their muscular activation and motion patterns. The results indicate differences in the activation patterns of the groups in the test task "floor to waist lift" common in many occupations. PMID- 16242307 TI - G protein betagamma11 complex translocation is induced by Gi, Gq and Gs coupling receptors and is regulated by the alpha subunit type. AB - G protein activation by Gi/Go coupling M2 muscarinic receptors, Gq coupling M3 receptors and Gs coupling beta2 adrenergic receptors causes rapid reversible translocation of the G protein gamma11 subunit from the plasma membrane to the Golgi complex. Co-translocation of the beta1 subunit suggests that gamma11 translocates as a betagamma complex. Pertussis toxin ADP ribosylation of the alphai subunit type or substitution of the C terminal domain of alphao with the corresponding region of alphas inhibits gamma11 translocation demonstrating that alpha subunit interaction with a receptor and its activation are requirements for the translocation. The rate of gamma11 translocation is sensitive to the rate of activation of the G protein alpha subunit. alpha subunit types that show high receptor activated rates of guanine nucleotide exchange in vitro support high rates of gamma11 translocation compared to alpha subunit types that have a relatively lower rate of guanine nucleotide exchange. The results suggest that the receptor induced translocation of gamma11 is controlled by the rate of cycling of the G protein through active and inactive forms. They also demonstrate that imaging of gamma11 translocation can be used as a non-invasive tool to measure the relative activities of wild type or mutant receptor and alpha subunit types in a live cell. PMID- 16242309 TI - Semi-automatic learning of simple diagnostic scores utilizing complexity measures. AB - OBJECTIVE: Knowledge acquisition and maintenance in medical domains with a large application domain ontology is a difficult task. To reduce knowledge elicitation costs, semi-automatic learning methods can be used to support the domain specialists. They are usually not only interested in the accuracy of the learned knowledge: the understandability and interpretability of the learned models is of prime importance as well. Then, often simple models are more favorable than complex ones. METHODS AND MATERIAL: We propose diagnostic scores as a promising approach for the representation of simple diagnostic knowledge, and present a method for inductive learning of diagnostic scores. It can be incrementally refined by including background knowledge. We present complexity measures for determining the complexity of the learned scores. RESULTS: We give an evaluation of the presented approach using a case base from the fielded system SonoConsult. We further discuss that the user can easily balance between accuracy and complexity of the learned knowledge applying the presented measures. CONCLUSIONS: We argue that semi-automatic learning methods can support the domain specialist efficiently when building (diagnostic) knowledge systems from scratch. The presented complexity measures allow for an intuitive assessment of the learned patterns. PMID- 16242310 TI - Concurrent presentations of the sarcoidosis, tuberculosis and lymphoma in a single patient. AB - A 56-year-old female patient, developing ALK-negative anaplastic T cell lymphoma 7 years after the diagnosis of sarcoidosis with clinical and histopathological features of tuberculosis has been presented. We herein present concurrent occurrence of the sarcoidosis, lymphoma and tuberculosis along with the confusing findings during the investigation for the establishing the diagnosis and management that represented a great challenge. PMID- 16242311 TI - Cigarette smoke extract modulates respiratory defence mechanisms through effects on T-cells and airway epithelial cells. AB - Chronic obstructive pulmonary disease (COPD) is a disease primarily caused by cigarette smoking, which in turn has been shown to affect the susceptibility to and progression of airway infections. The question addressed in this study was how components from cigarette smoke could affect the defence mechanisms of T cells and epithelial cells, and thereby contribute to the development of the COPD pathology. T-cells and monocytes were isolated from buffycoats from healthy donors and T-cell responses studied in response to cigarette smoke extract (CSE). Activation level (CD25 expression), proliferation (BrdU incorporation) and intracellular expression of the cytotoxic markers granzyme-b and TIA-1 were determined using flowcytometry. Normal human bronchial epithelial cells were obtained from Cambrex and differentiated in air-liquid interface cultures. After exposure to CSE barrier function (trans-epithelial electric resistance, TEER), MUC5AC and interleukin-8 production were measured. T-cell activation, proliferation and expression of the cytotoxic proteins granzyme-b and TIA-1 were significantly reduced in response to 0.5-1% of CSE. The epithelial cells were more resistant to CSE and responded at doses 20 times higher than T-cells. The expression of interleukin-8 and MUC5AC was significantly increased after exposure to 15% and 30% CSE and TEER was largely unaffected at 30% CSE but clearly reduced at 40% CSE. This study shows that mechanisms, in both T-cells and airway epithelial cells, involved in the defence against infectious agents are modulated by CSE. PMID- 16242312 TI - An audit into the efficacy of single use bacterial/viral filters for the prevention of equipment contamination during lung function assessment. AB - Lung function testing has been suggested to provide a potential risk regarding cross-infection between patients. About 155 patients (86 infectious, 69 non infectious) used a single use bacterial/viral filter when performing routine lung function tests. Swabs from the patient side of the filter (Proximal) and the equipment side (Distal), and two sections of the filter itself were cultured. About 33/155 samples showed bacterial growth on the Proximal compared with 2/155 on the Distal side (P<0.01). Growth was obtained from the filter in 125/155 (80.6%) of cases. Pathogenic micro-organisms such as Pseudomonas aeruginosa (4 cases) and Staphylococcus aureus (5 cases) were isolated. Appropriate infection control measures should be used when performing lung function tests. PMID- 16242313 TI - Neuronal differentiation in C. elegans. AB - The small size and defined connectivity of the C. elegans nervous system and the amenability of this species to systematic functional screens have continued to yield new insights into neuronal differentiation. Many aspects of C. elegans neuronal development resemble those of other more complex neurons. The basic cellular machinery of synaptic transmission is highly conserved. Recent work has begun to unveil the roles of proteoglycans in axon guidance and branching, and of the extracellular matrix in neuronal process maintenance. The importance of ubiquitin-mediated protein turnover in neuronal differentiation is revealed by the identification of new and conserved pathways that promote the organization and function of the synapse. PMID- 16242315 TI - Hospital variation in referral for primary radiotherapy in South Netherlands, 1988-1999. AB - In this study, we have assessed whether referral for primary radiotherapy varied according to hospital size in a region with 1 million inhabitants served by community hospitals. We studied 20178 patients diagnosed with breast, non-small cell lung, prostate, rectal, or endometrial cancer between 1988 and 1999. We used logistic regression analysis, adjusted for age, stage and period of diagnosis. Medium-sized and small hospitals referred breast cancer patients more often (OR=2.2, 95%CI: 2.0-2.5, OR=1.2, 95%CI: 1.1-1.4, respectively), and patients with prostate cancer less often (OR=0.7 (0.5-0.8) and 0.7 (0.6-0.9), respectively). Referral rates for patients with non-small cell lung and rectal cancer showed minor differences according to hospital size, referral for endometrial cancer was somewhat higher for patients from medium-sized hospitals (OR=1.5 (1.0-2.1)). Time trends in variation were shown, but differences according to hospital size only decreased over time for rectal cancer. Despite multidisciplinary oncology meetings and treatment guidelines there were large variations in rates of referral for radiotherapy. PMID- 16242314 TI - Phenolic acids from beer are absorbed and extensively metabolized in humans. AB - In spite of the wide literature describing the biological effects of phenolic compounds, scarce data are available on their absorption from diet. In the present work, we studied the absorption in humans of phenolic acids from beer, a common beverage rich in different phenolic acids with related chemical structures. Beer was analyzed for free and total (free+bound) phenolic acids. Ferulic, caffeic and sinapic acids were present in beer mainly as bound forms, while 4-hydroxyphenylacetic acid and p-coumaric acid were present mainly as free forms. Vanillic acid was present equally in the free and bound forms. Plasma samples were collected before and 30 and 60 min after beer administration and analyzed for free and conjugated phenolic acid content. A significant two- to fourfold increase in plasma levels of phenolic acids was detected with peak concentrations at 30 min after beer ingestion. 4-Hydroxyphenylacetic acid was present in plasma mainly as nonconjugated forms while p-coumaric acid was present equally as nonconjugated and conjugated forms. Ferulic, vanillic and caffeic acids were present in plasma predominantly as conjugated forms, with a slight prevalence of sulfates with respect to glucuronates. Our results indicate that phenolic acids from beer are absorbed from the gastrointestinal tract and are present in blood after being largely metabolized to the form of glucuronide and sulfate conjugates. The extent of conjugation is related to the chemical structure of phenolic acids: the monohydroxy derivatives showing the lowest conjugation degree and the dihydroxy derivatives showing the highest one. PMID- 16242316 TI - Combined Pap and HPV testing in primary screening for cervical abnormalities: should HPV detection be delayed until age 35? AB - In 2003, the United States Food and Drug Administration has approved the Hybrid Capture 2 assay for use with a Pap test to adjunctively screen women of 30 years and older for the presence of high-risk human papillomavirus (HR-HPV) infection. Although the predictive power of a negative test is strong, the number of false positives may still be high. We investigated HPV prevalence in relation to age in a group of 2293 women, aged between 20 and 50, with normal cytology. Overall HR HPV prevalence was 6.9% (95%CI=5.9-8.0%). Regression analysis using 5-year intervals showed that the HR-HPV prevalence did not significantly decline up to age 34, whereas it declined significantly after age 35. This would suggest that postponing HPV detection in primary screening from age 30 to 35 would result in a decrease of almost 50% of the number of women with normal cytology and a transient HPV infection. However, larger scale studies are required to confirm this finding. PMID- 16242317 TI - Cloning, restriction mapping and phylogenetic relationship of genomic components of MYMIV from Lablab purpureus. AB - The present work describes cloning of genomic components of whitefly transmitted geminivirus infecting Lablab purpureus syn. Dolichos lablab (commonly known as Dolichos bean or Hyacinth bean). The genome characterization using PCR with geminiviral degenerate primers and DNA sequencing were used to describe the bipartite virus associated with yellow mosaic disease of Dolichos lablab. Full length DNA-A and DNA-B clones were obtained. The DNA-A sequence analysis showed that the isolate was similar to other Mungbean yellow mosaic India virus (MYMIV) isolates reported earlier. The nucleotide sequence analysis of the full-length DNA-A of virus isolate revealed more than 97% homology with Mungbean yellow mosaic India virus-[Cowpea] (AF481865), while the DNA-B also showed >95% homology with MYMIV-[Cp] (AF503580) and MYMIV-[Sb] (AY049771). The phylogenetic analysis of present isolate showed close relationship to legume geminiviruses. The nucleotide sequence analysis showed presence of six open reading frames (ORFs) in DNA-A, with 2 ORFs aligned in sense and 4 ORFs in antisense orientation. Similarly, DNA-B contained two open reading frames (ORFs), one in sense and another in antisense orientation. PMID- 16242318 TI - Xylitol production from corn fiber and sugarcane bagasse hydrolysates by Candida tropicalis. AB - A natural isolate, Candida tropicalis was tested for xylitol production from corn fiber and sugarcane bagasse hydrolysates. Fermentation of corn fiber and sugarcane bagasse hydrolysate showed xylose uptake and xylitol production, though these were very low, even after hydrolysate neutralization and treatments with activated charcoal and ion exchange resins. Initial xylitol production was found to be 0.43 g/g and 0.45 g/g of xylose utilised with corn fiber and sugarcane bagasse hydrolysate respectively. One of the critical factors for low xylitol production was the presence of inhibitors in these hydrolysates. To simulate influence of hemicellulosic sugar composition on xylitol yield, three different combinations of mixed sugar control experiments, without the presence of any inhibitors, have been performed and the strain produced 0.63 g/g, 0.68 g/g and 0.72 g/g of xylose respectively. To improve yeast growth and xylitol production with these hydrolysates, which contain inhibitors, the cells were adapted by sub culturing in the hydrolysate containing medium for 25 cycles. After adaptation the organism produced more xylitol 0.58 g/g and 0.65 g/g of xylose with corn fiber hydrolysate and sugarcane bagasse hydrolysate respectively. PMID- 16242319 TI - Bioconversion of industrial wastewater and wastewater sludge into Bacillus thuringiensis based biopesticides in pilot fermentor. AB - Starch industry wastewater (SWW), slaughterhouse wastewater (SHWW) and secondary sludges from three different wastewater treatment plants (Jonquiere--JQS, Communaute Urbaine de Quebec--CUQS and Black lake-BLS) were used as raw materials for the production of Bacillus thuringiensis (Bt) based biopesticides in a pilot scale fermentor (100 L working volume). The slaughterhouse wastewater exhibited the lowest Bt growth and entomotoxcity (Tx) potential (measured against spruce budworm) due to low availability of carbon, nitrogen and other nutrients. Performance variation (growth, sporulation, proteolytic activity and Tx potential) within the three types of sludges was directly related to the availability of nitrogen and carbohydrates, which could change with sludge origin and methods employed for its generation. The Tx potential of Bt obtained in different secondary sludges (JQS: 12 x 10(9) SBU/L; CUQS: 13 x 10(9) SBU/L and BLS: 16 x 10(9) SBU/L) and SWW (18 x 10(9) SBU/L) was higher than the soybean based synthetic medium (10 x 10(9) SBU/L). The maximum protease activity was obtained in CUQ secondary sludge (4.1 IU/mL) due to its high complex protein concentration. Nevertheless, high carbohydrate concentration in SWW repressed enzyme production. The secondary sludges and SWW were found to be suitable raw materials for high potency Bt biopesticide production. PMID- 16242320 TI - A protease stable in organic solvents from solvent tolerant strain of Pseudomonas aeruginosa. AB - A solvent tolerant strain of Pseudomonas aeruginosa (PseA) was isolated from soil samples by cyclohexane enrichment in medium. The strain was able to sustain and grow in a wide range of organic solvents. The adaptation of P. aeruginosa cell towards solvents was seen at membrane level in transmission electron micrographs. It also secreted a novel protease, which exhibited remarkable solvent stability and retained most of the activity at least up to 10 days in the presence of hydrophobic organic solvents (log P > or = 2.0) at 25% (v/v) concentrations. The protease was able to withstand as high as 75% concentration of solvents at least up to 48 h. P. aeruginosa strain and its protease, both seem promising for solvent bioremediation, wastewater treatment and carrying out biotransformation in non-aqueous medium. PMID- 16242321 TI - Aza-analogues of the marine pyrroloquinoline alkaloids wakayin and tsitsikammamines: synthesis and topoisomerase inhibition. AB - Two aza-analogues of the marine pyrroloquinoline alkaloids wakayin and tsitsikammamines A and B have been synthesized. The strategy used was based on a 1,3-dipolar cycloaddition reaction between indole 4,7-dione and a diazo aminopropane derivative. One of the two analogues partially inhibits human topoisomerase I, whereas synthetic intermediates inhibit the enzyme DNA cleavage activity at a concentration comparable to that of the control drug camptothecin. PMID- 16242322 TI - p38 MAP kinase inhibitors: metabolically stabilized piperidine-substituted quinolinones and naphthyridinones. AB - Quinolinones and naphthyridinones with C7 N-t-butyl piperidine substituents were found to be potent p38 MAP kinase inhibitors. These compounds significantly suppress TNF-alpha release in both cellular and LPS-stimulated whole blood assays. They also displayed excellent PK profiles across three animal species. Quinolinone at 10 mpk showed comparable oral efficacy to that of dexamethasone at 1 mpk in a murine collagen-induced arthritis model. PMID- 16242323 TI - Synthesis and biological activity of novel 4-phenyl-1,8-naphthyridin-2(1H)-on-3 yl ureas: potent acyl-CoA:cholesterol acyltransferase inhibitor with improved aqueous solubility. AB - 4-Aryl-1,8-naphthyridin-2(1H)-on-3-yl urea derivatives with hydrophilic groups were synthesized in order to improve aqueous solubility and pharmacokinetic property. SMP-797 possessing (4-aminophenyl)ureido and 3-(hydroxypropoxyphenyl) moieties showed potent ACAT inhibitory activity and excellent oral efficacy. PMID- 16242324 TI - Safrole oxide induces apoptosis by activating caspase-3, -8, and -9 in A549 human lung cancer cells. AB - Previously we found that 3,4-(methylenedioxy)-1-(2',3'-epoxypropyl)-benzene (safrole oxide) induced a typical apoptosis in A549 human lung cancer cells. In this study, we further investigated which caspases were activated by safrole oxide during the apoptosis. The data showed that the activity of caspase-3, -8, and -9 was significantly enhanced by the compound, which suggested that safrole oxide might be used as a caspase promoter to initiate lung cancer cell apoptosis. PMID- 16242325 TI - The design, synthesis, and evaluation of two universal doxorubicin-linkers: preparation of conjugates that retain topoisomerase II activity. AB - The design, synthesis, and evaluation of two N-alkylmaleimide aldehydes have been achieved, which upon reductive alkylation with the C3'-amino group of doxorubicin (DOX) permits the preparation of DOX conjugates via Michael addition of thiol containing vectors. This method enables the mild, facile, and high-throughput preparation of DOX conjugates that retain the basic C3'-nitrogen, a pre-requisite for topoisomerase II inhibition. Seven DOX-amino acid conjugates were prepared, each displaying similar inhibitory activity as the parent drug. PMID- 16242326 TI - Synthesis of the PPARbeta/delta-selective agonist GW501516 and C4-thiazole substituted analogs. AB - Sequential, position-selective, Pd-catalyzed cross-coupling reactions of 2,4 dibromo-5-hydroxymethylthiazole provided the scaffold for the synthesis of GW501516, the most potent PPARbeta/delta agonist yet described, and equally selective analogs at the thiazole-C4 position. PMID- 16242327 TI - Crystal structure of human ERK2 complexed with a pyrazolo[3,4-c]pyridazine derivative. AB - A series of pyrazolopyridazine compounds were briefly investigated as ERK2 inhibitors. The crystal structure of ERK2 complexed with an allyl derivative was determined. The compound induces structural change including movement of the glycine-rich loop and peptide flip between Met108-Glu109. As a result, the newly formed subsite can recognize small hydrophobic substituents but not hydrophilic ones. PMID- 16242328 TI - Synthesis and biological evaluation of 3-ethylidene-1,3-dihydro-indol-2-ones as novel checkpoint 1 inhibitors. AB - Chk1 inhibitors have emerged as a novel class of neoplastic agents for abrogating the G2 DNA damage checkpoint arrest. Analogs of the Chk1 inhibitor, 3-ethylidene 1,3-dihydro-indol-2-one, were synthesized and tested in vitro for their inhibitory activities. The most promising compound identified from this series is analog 28, which possesses potent enzymatic and cellular activities. PMID- 16242329 TI - N-Hydroxyurea as zinc binding group in matrix metalloproteinase inhibition: mode of binding in a complex with MMP-8. AB - The first crystallographic structure of an N-hydroxyurea inhibitor bound into the active site of a matrix metalloproteinase is reported. The ligand and three other analogues were prepared and studied as inhibitors of MMP-2, MMP-3, and MMP-8. The crystal structure of the complex with MMP-8 shows that the N-hydroxyurea, contrary to the analogous hydroxamate, binds the catalytic zinc ion in a monodentate rather than bidentate mode and with high out-of-plane distortion of the amide bonds. PMID- 16242331 TI - Intraoperative electron beam radiotherapy (ELIOT) to the breast: a need for a quality assurance programme. AB - Intraoperative radiotherapy (IORT) is a technique in which a high, single fraction radiation dose is delivered directly to the tumour bed during a surgical intervention, after the removal of a neoplastic mass. IORT has been recently used in early stage cancer as an exclusive radiation modality, rather than as a boost, especially for breast tumours, in particular at the European Institute of Oncology in Milan, where the technique has been called electron intraoperative therapy (ELIOT). Our studies on more than 1000 patients have demonstrated the feasibility of the technique and it is expected that its application will become more widespread in the immediate future. It is important to emphasise that ELIOT relies not only on new technological developments, but also on a multidisciplinary team with clear roles and responsibilities, the establishment of a programme of quality assurance with appropriate guidelines and a comprehensive staff development programme. PMID- 16242330 TI - Design and synthesis of downsized metastin (45-54) analogs with maintenance of high GPR54 agonistic activity. AB - Metastin has been identified as a metastasis suppressor gene product that mediates its function through a G protein coupled receptor, GPR54. To refine insight into the critical pharmacophore for the activation of GPR54, we have conducted alanine and d-amino acid scanning on a biologically active metastin fragment (45-54). Based on these data and structures of peptides previously reported to activate GPR54, a series of shortened metastin (45-54) derivatives were synthesized and tested for the ability to induce GPR54 signaling. These biological experiments were performed in yeast containing human GPR54 that was coupled to the pheromone response pathway and a pheromone responsive lacZ reporter gene. Compounds 32, 33, and 39, which possess an N-terminal basic group and a C-terminal RW-amide motif, were strong agonists, similar to the level of metastin. This may provide an approach to reverse the pro-metastatic effect of metastin deletion in multiple malignant tumors. PMID- 16242332 TI - Staphylococcal innate immune evasion. AB - Upon entering the human body, bacteria are confronted with the sophisticated innate defense mechanisms of the human host. From work in recent years it has become obvious that a new and growing family of small and excreted proteins can counteract the antibacterial effects of innate immunity. These highly selective proteins pick out crucial elements of our immune system and inhibit their function. In Staphylococcus aureus these proteins act on specific cellular receptors, on antimicrobial peptides and especially on the complement system. The combined action of this growing group of essential virulence factors ascertains efficient innate immune evasion. PMID- 16242333 TI - Tibial plateau fracture following gracilis-semitendinosus anterior cruciate ligament reconstruction: The tibial tunnel stress-riser. AB - Tibial plateau fractures following anterior cruciate ligament (ACL) reconstruction are extremely rare. This is the first reported case of a tibial plateau fracture following four-strand gracilis-semitendinosus autograft ACL reconstruction. The tibial tunnel alone may behave as a stress riser which can significantly reduce bone strength. PMID- 16242334 TI - Solid phase combinatorial synthesis of benzothiazoles and evaluation of topoisomerase II inhibitory activity. AB - To investigate one possible mechanism of action of the cytotoxic activity of benzothiazoles, we synthesized 2-(substituted-phenyl)benzothiazoles and evaluated their ability to inhibit topoisomerase II activities. Solid phase combinatorial method using trityl resin was employed and benzothiazole derivatives with various substitution on 2'-, 3'-, or 4'-position of phenyl group were obtained in ca. 30 mg scale (7-96% yield). Most of the compounds synthesized exhibited topoisomerase II inhibitory activity at 100 microM. 2-(3-Amino-4-methylphenyl)benzothiazole showed high activity (IC(50) = 71.7 microM), comparable to etoposide (IC(50) = 78.4 microM). PMID- 16242335 TI - Guanaconetins, new antitumoral acetogenins, mitochondrial complex I and tumor cell growth inhibitors. AB - The antitumoral activity of a series of acetylated bis-tetrahydrofuranic acetogenins with a threo/trans/threo/trans/erythro relative configuration was characterized by four new natural and two semisynthetic, 15,24,30-trioxygenated acetogenins that were found to inhibit mitochondrial complex I enzyme as well as growth of several tumor cell lines. Placement of acetyl groups along the alkyl chain modulated the potency of the bis-tetrahydrofuranic acetogenins and could be important for future utilization of these compounds as chemotherapeutic agents. PMID- 16242336 TI - Microwave enhanced solid support synthesis of fluorine containing benzopyrano triazolo-thiadiazepines as potent anti-fungal agents. AB - A facile, dry media procedure has been developed for the synthesis of a series of a new class of fluorine containing 3-alkyl-7-chloro-11a,12-dihydro-11-phenyl-12 (substituted aryl)-11H-benzopyrano[4,3-e][1,2,4]-triazolo[3,4-b][1,3,4] thiadiazepines (4a-k) under microwaves using basic alumina as solid support. The reaction time has been brought down from hours (60-65 h) to minutes (3-5 min) with improved yield as compared to conventional method, demonstrating the versatility of the process. The method reported herein is devoid of the hazards of solution-phase reactions. The synthesized compounds have been screened 'in vitro' for anti-fungal activity against Rhizoctonia solani, Fusarium oxysporum and Collectotrichum capsici. Most of the compounds have shown good activity against these pathogens. PMID- 16242337 TI - Synthesis, antiviral, and antitumor activity of 2-substituted purine methylenecyclopropane analogues of nucleosides. AB - The Z- and E-2-fluoro- and 2-chloropurine methylenecyclopropanes 9a,b and 10a,b as well as enantiomeric Z-isoguanine methylenecyclopropanes 11a,b and their phenyl phosphoralaninate pronucleotides 11c,d were synthesized and their antiviral activity against several viruses was evaluated. Fluoro analogues 9a and 10a were active against human cytomegalovirus but they were cytotoxic at approximately the same concentrations. Chloro derivatives 9b and 10b were non cytotoxic and effective against Epstein-Barr virus in Daudi cells. Isoguanine analogues 11a-d lacked antiviral activity but pronucleotides 11c,d were substrates for porcine liver esterase. From the group of 9a,b and 10a,b, the fluoro analogues 9a and 10a exhibited antitumor activity but only the Z-isomer 9a had a selective effect. PMID- 16242338 TI - Synthesis and in vitro activities of ferrocenic aminohydroxynaphthoquinones against Toxoplasma gondii and Plasmodium falciparum. AB - Fourteen ferrocenyl aminohydroxynaphthoquinones, analogues of atovaquone, were synthesized from the hydroxynaphthoquinone core. These novel atovaquone derivatives were tested for their in vitro activity against two apicomplexan parasites of medical importance, Toxoplasma gondii and Plasmodium falciparum, including resistant strains to atovaquone (T. gondii) and chloroquine (P. falciparum). Three of these ferrocenic atovaquone derivatives composed of the hydroxynaphthoquinone core plus an amino-ferrocenic group and an aliphatic chain with 6-8 carbon atoms were found to be significantly active against T. gondii. Moreover, these novel compounds were also effective against the atovaquone resistant strain of T. gondii (Ato(R)). PMID- 16242339 TI - Thioureido N-acetyllactosamine derivatives as potent galectin-7 and 9N inhibitors. AB - Derivatives of N-acetyllactosamine carrying structurally diverse thioureido groups at galactose C3 were prepared from a C3'-azido N-acetyllactosamine derivative in a three-step reaction sequence involving azide reduction and isothiocyanate formation by thiophosgene treatment of the C3-amine, followed by reaction of the isothiocyanate with a panel of amines. Evaluation of the N acetyllactosamine thioureas as inhibitors against galectins-1, 3, 7, 8N (N terminal domain), and 9N (N-terminal domain) revealed thiourea-mediated affinity enhancements for galectins-1, 3, 7, and 9N. In particular, good inhibitors were discovered against galectin-7 and 9N (K(d) 23 and 47 microM, respectively, for a 3-pyridylmethylthiourea derivative), which represents more than an order of magnitude affinity enhancement over the parent natural N-acetyllactosamine. PMID- 16242340 TI - Novel 5-(2-hydroxyphenyl)-3-substituted-2,3-dihydro-1,3,4-oxadiazole-2-thione derivatives: promising anticancer agents. AB - A series of 5-(2-hydroxyphenyl)-3-substituted-2,3-dihydro-1,3,4-oxadiazole-2 thione derivatives was synthesized and 13 of them were selected by the National Cancer Institute (NCI) and evaluated for their in vitro anticancer activity. Seven of the investigated compounds, 3i, 3j, 3k, 3o, 3p, 3q, and 3r, displayed high anticancer activity in the primary assay. These compounds have been selected for a full anticancer screening against a 60-cell panel assay where they showed non-selective broad spectrum and promising activity against all cancer cell lines. Compounds 3j and 3k proved to be the active members in this study compared to 5-fluorouracil and cyclophosphamide as reference drugs, respectively. Compounds 3j and 3k were identified as promising lead compounds. PMID- 16242341 TI - Bone microarchitecture evaluated by histomorphometry. AB - The increasing use of densitometric devices for assessing bone fragility has progressively strengthened the assumption that mass is the most important property determining bone mechanical competence. Nevertheless, structure and microarchitecture are relevant aspects of bone strength. The study of microarchitecture is based on the measure of width, number, and separation of trabeculae as well as on their spatial organization. There are several methods to assess bone architecture, particularly at the trabecular level. In particular, histomorphometry, based on the use of optical microscopy and on the principles of quantitative histology and stereology, evaluates microarchitecture two dimensionally, even if these measures appear well correlated to the three dimensional structure and properties of bone. In addition, new computerized methods allow the acquisition of more sophisticated measurements by means of a digitizer have been introduced to integrate the use of the microscope. These methods supply information on trabecular width as well as on its distribution and on the organization of the trabeculae in the marrow space. Microarchitecture seems to be a determinant of bone fragility independent of bone density and it is important for understanding the mechanisms of bone fragility as well as the action of the drugs used to prevent osteoporotic fractures. Several in vivo studies (on animals and humans) can provide an additional interpretation for the anti-fracture effect of such drugs. For instance, bisphosphonates and parathyroid hormone seem to preserve or even improve microarchitecture. The challenge for the future will be to evaluate bone quality in vivo with the same or better resolution and accuracy than the invasive methods used today. PMID- 16242342 TI - Siliceous spicules in marine demosponges (example Suberites domuncula). AB - All metazoan animals comprise a body plan of different complexity. Since- especially based on molecular and cell biological data--it is well established that all metazoan phyla, including the Porifera (sponges), evolved from a common ancestor the search for common, basic principles of pattern formation (body plan) in all phyla began. Common to all metazoan body plans is the formation of at least one axis that runs from the apical to the basal region; examples for this type of organization are the Porifera and the Cnidaria (diploblastic animals). It seems conceivable that the basis for the formation of the Bauplan in sponges is the construction of their skeleton by spicules. In Demospongiae (we use the model species Suberites domuncula) and Hexactinellida, the spicules consist of silica. The formation of the spicules as the building blocks of the skeleton, starts with the expression of an enzyme which was termed silicatein. Spicule growth begins intracellularly around an axial filament composed of silicatein. When the first layer of silica is made, the spicules are extruded from the cells and completed extracellularly to reach their the final form and size. While the first steps of spicule formation within the cells are becoming increasingly clear, it remains to be studied how the extracellularly present silicatein strings are formed. The understanding of especially this morphogenetic process will allow an insight into the construction of the amazingly diverse skeleton of the siliceous sponges; animals which evolved between two periods of glaciations, the Sturtian glaciation (710-680 MYA) and the Varanger-Marinoan ice ages (605-585 MYA). Sponges are--as living fossils--witnesses of evolutionary trends which remained unique in the metazoan kingdom. PMID- 16242344 TI - Immune- and enzyme histochemical characterisation of leukocyte populations within lymphoid and mucosal tissues of Atlantic halibut (Hippoglossus hippoglossus). AB - Leukocyte populations within the kidney, spleen, posterior intestine and gills of Atlantic halibut were investigated using a panel of histological, enzyme- and immunohistochemical methods. In the kidney and spleen, a diverse population of leukocytes was associated with the extensive network of sinusoids and larger blood vessels present in these tissues. IgM+ cells (B-cells, plasma cells and IgM bearing macrophages) and large mononuclear cells showing reactivity for non specific esterase (NSE) and acid phosphatase (ACP), representing macrophage populations, were often associated with vessel walls that were also the site of trapping of fluorescent microspheres. In the kidney, trapping of 0.1 and 0.5 microm diameter microspheres occurred at these sites but in the spleen, the 0.1 microm diameter microspheres were retained in ellipsoids. The lymphoid tissues of the kidney and spleen possessed a spread population of 5'-nucleotidase+ (5'N+) cells but compartmentalisation of the splenic white pulp was suggested by an absence of these 5'N+ reticular cells in areas associated with melanomacrophage accumulations and in areas rich in IgM+ cells. A striking feature of the mucosal tissues was the diversity of leukocyte populations within the epithelium particularly of the posterior intestine, including IgM+ cells and NSE+, ACP+ and 5'N+ mononuclear cells. Although limited in numbers in the posterior intestine, IgM+ cells were more common in the epithelium than in the lamina propria. In the gills, leukocytes as detected by enzymatic reactivity were scarce, but IgM+ cells were very abundant in the stratified epithelium of the gill arch and filaments. The difference in distribution of these leukocyte populations between the intestines and gills suggested a compartmentalisation of the mucosal immune system and the need to assess the immunological competence of mucosal tissues in Atlantic halibut. PMID- 16242343 TI - Helical crystallization on nickel-lipid nanotubes: perfringolysin O as a model protein. AB - To facilitate purification and subsequent structural studies of recombinant proteins the most widely used genetically encoded tag is the histidine tag (His tag) which specifically binds to N-nitrilotriacetic-acid-chelated nickel ions. Lipids derivatized with a nickel-chelating head group can be mixed with galactosylceramide glycolipids to prepare lipid nanotubes that bind His-tagged proteins. In this study, we use His-tagged perfringolysin O (PFO), a soluble toxin secreted by the bacterial pathogen Clostridium perfringens, as a model protein to test the utility of nickel-lipid nanotubes as a tool for structural studies of His-tagged proteins. PFO is a member of the cholesterol dependent cytolysin family (CDC) of oligomerizing, pore-forming toxins found in a variety of Gram-positive bacterial pathogens. CDC pores have been difficult to study by X ray crystallography because they are membrane associated and vary in size. We demonstrate that both a wild-type and a mutant form of PFO form helical arrays on nickel-lipid containing nanotubes. Cryo-electron microscopy and image analysis of the helical arrays were used to reconstruct a 3D density map of wild-type PFO. This study suggests that the use of nickel-lipid nanotubes may offer a general approach for structural studies of recombinant proteins and may provide insights into the molecular interactions of proteins that have a natural affinity for a membrane surface. PMID- 16242345 TI - Classification of large array surface myoelectric potentials from subjects with and without low back pain. AB - An algorithm was developed and tested for differentiating between the spatial distribution of large arrays of surface electromyographic (LASE) data from subjects with and without low back pain (LBP). The surface EMG data from 62 channels were collected from the low back of 161 healthy and 44 acute (less than 6-weeks) LBP subjects in three minimum stress postural positions including standing, 20 degrees of trunk flexion (at hip joint) and standing with arms extended forward holding a 1.36kg (3lb) weight in each hand. These data were statistically analyzed and the spatial distribution of the root mean square (RMS) values was used in a multivariate quadratic discriminant model to reclassify the healthy and acute LBP subjects. The most predictive results were obtained from the 'flexion' group of experiments and correctly reclassified 95.5% (42/44) of the acute LBP subjects and 99.4% (160/161) of the healthy subjects. The success rate of this reclassification based on surface distribution of myoelectric potentials was found to be better than the reported patient classifications based on a smaller set of electrode pairs using fewer subjects [Peach JP, McGill SM, Classification of low back pain with use of spectral electromyogram parameters. Spine 23(10):1998;1117-23; Roy SH, De Luca CJ, Emley M, Oddsson LI, Buijs RJ, Levins JA, Newcombe DS, Jabre JF. Classification of back muscle impairment based on the surface electromyographic signal. J Rehabil Res Dev 34(4):1997;405-14 [review]]. The results indicated the potential of the model for clinical patient classification. PMID- 16242346 TI - Combining EEG and fMRI to investigate the post-movement beta rebound. AB - The relationship between synchronous neuronal activity as measured with EEG and the blood oxygenation level dependent (BOLD) signal as measured during fMRI is not clear. This work investigates the relationship by combining EEG and fMRI measures of the strong increase in beta frequency power following movement, the so-called post-movement beta rebound (PMBR). The time course of the PMBR, as measured by EEG, was included as a regressor in the fMRI analysis, allowing identification of a region of associated BOLD signal increase in the sensorimotor cortex, with the most significant region in the post-central sulcus. The increase in the BOLD signal suggests that the number of active neurons and/or their synaptic rate is increased during the PMBR. The duration of the BOLD response curve in the PMBR region is significantly longer than in the activated motor region, and is well fitted by a model including both motor and PMBR regressors. An intersubject correlation between the BOLD signal amplitude associated with the PMBR regressor and the PMBR strength as measured with EEG provides further evidence that this region is a source of the PMBR. There is a strong intra subject correlation between the BOLD signal amplitude in the sensorimotor cortex during movement and the PMBR strength as measured by EEG, suggesting either that the motor activity itself, or somatosensory inputs associated with the motor activity, influence the PMBR. This work provides further evidence for a BOLD signal change associated with changes in neuronal synchrony, so opening up the possibility of studying other event-related oscillatory changes using fMRI. PMID- 16242347 TI - Improved BOLD detection in the medial temporal region using parallel imaging and voxel volume reduction. AB - Using gradient-echo EPI, signal dropout due to macroscopic off resonance effects can prevent blood-oxygenation-level-dependent (BOLD) signal change detection. The anterior medial temporal lobe (MTL) is located near these susceptibility gradients and therefore shows considerable signal dropout with GE-EPI. Reducing the volume of the image voxel reduces susceptibility-related signal dropout. However, this is accompanied by a prohibitive reduction in signal-to-noise ratio (SNR). To compensate for SNR loss with smaller voxels, we used a multi-channel MRI receiver with an array of receive-only 16-element surface coils at 3 T. We demonstrate that the reduction of susceptibility artifacts, through use of high resolution images, coupled with the gains in image SNR from the array coil improves the temporal signal-to-noise ratio (TSNR) and enhances the contrast-to noise ratio (CNR). Furthermore, a comparison of 2 mm with 4-mm-thick axial images both with the same in-plane resolution showed that thinner slices enhanced TSNR and CNR throughout the ventral-medial regions of the temporal lobes, with the greatest improvement in the most anterior regions of the MTL. Further improvements were seen when adjacent 2 mm slices were combined to match overall voxel volume. These results demonstrate that BOLD investigation of anterior MTL function can be enhanced by decreasing voxel size but only in combination with the SNR gained by using the 16-channel head coil system. PMID- 16242348 TI - A multivariate approach for processing magnetization effects in triggered event related functional magnetic resonance imaging time series. AB - Triggered event-related functional magnetic resonance imaging requires sparse intervals of temporally resolved functional data acquisitions, whose initiation corresponds to the occurrence of an event, typically an epileptic spike in the electroencephalographic trace. However, conventional fMRI time series are greatly affected by non-steady-state magnetization effects, which obscure initial blood oxygen level-dependent (BOLD) signals. Here, conventional echo-planar imaging and a post-processing solution based on principal component analysis were employed to remove the dominant eigenimages of the time series, to filter out the global signal changes induced by magnetization decay and to recover BOLD signals starting with the first functional volume. This approach was compared with a physical solution using radiofrequency preparation, which nullifies magnetization effects. As an application of the method, the detectability of the initial transient BOLD response in the auditory cortex, which is elicited by the onset of acoustic scanner noise, was used to demonstrate that post-processing-based removal of magnetization effects allows to detect brain activity patterns identical with those obtained using the radiofrequency preparation. Using the auditory responses as an ideal experimental model of triggered brain activity, our results suggest that reducing the initial magnetization effects by removing a few principal components from fMRI data may be potentially useful in the analysis of triggered event-related echo-planar time series. The implications of this study are discussed with special caution to remaining technical limitations and the additional neurophysiological issues of the triggered acquisition. PMID- 16242349 TI - In vivo diffusion tensor imaging (DTI) of brain subdivisions and vocal pathways in songbirds. AB - The neural substrate for song behavior in songbirds, the song control system (SCS), is thus far the best-documented brain circuit in which to study neuroplasticity and adult neurogenesis. Not only does the volume of the key song control nuclei change in size, but also the density of the connections between them changes as a function of seasonal and hormonal influences. This study explores the potentials of in vivo Diffusion-Tensor MRI (DT-MRI or DTI) to visualize the distinct, concentrated connections of the SCS in the brain of the starling (Sturnus vulgaris). In vivo DTI on starling was performed on a 7T MR system using sagittal and coronal slices. DTI was accomplished with diffusion gradients applied in seven non-collinear directions. Fractional Anisotropy (FA) maps allowed us to distinguish most of the grey matter and white matter-tracts, including the laminae subdividing the avian telencephalon and the tracts connecting the major song control nuclei (e.g., HVC with RA and X). The FA-maps also allowed us to discern a number of song control, auditory and visual nuclei. Fiber tracking was implemented to illustrate the discrimination of all tracts running from and to RA. Because of the remarkable plasticity inherent to the songbird brain, the successful implementation of DTI in this model could represent a useful tool for the in vivo exploration of fiber degeneration and regeneration and the biological mechanisms involved in brain plasticity. PMID- 16242350 TI - On why the elderly have normal semantic retrieval but deficient episodic encoding: a study of left inferior frontal ERP activity. AB - Age-related left inferior prefrontal cortex (LIPFC) blood flow reductions during semantic retrieval are associated with reduced subsequent episodic recognition memory performance but are inconsistent with age-invariant semantic retrieval performance. Therefore, we compared brain activity in young and elderly persons during low- and high-selection versions of a semantic task using ERPs recorded at 62 scalp locations. In an early interval (400-800 ms), both age groups showed more negativity over left inferior frontal scalp while performing the high- compared to the low-selection task. This early semantic selection-related negativity was associated with age-equivalent accuracy and reaction time on the semantic tasks. Further, in the early time interval, the ERPs of the young were more negative than those of the elderly. In addition, only the young showed a late selection-related negativity (1,200--1,400 ms). These age-related differences in left frontal ERP activity were associated with significant decrements in subsequent recognition for the elderly. Moreover, additional analyses revealed that larger amounts of negativity over left inferior frontal scalp locations during semantic retrieval were correlated with enhanced subsequent episodic recognition in both groups. In sum, the data suggest that (1) semantic retrieval and episodic encoding processes overlap temporally in similar brain regions, most likely including the LIPFC, and (2) the failure to recruit these regions in the service of episodic encoding underlies the age-related deficit in episodic memory. PMID- 16242351 TI - Impact of language proficiency and orthographic transparency on bilingual word reading: an fMRI investigation. AB - The purpose of the present functional magnetic resonance imaging (fMRI) investigation was to examine how language proficiency and orthographic transparency (letter-sound mapping consistency) modulate neural activity during bilingual single word reading. Spanish-English bilingual participants, more fluent in their second language (L2; English) than their native language (L1; Spanish), were asked to read words in the two languages. Behavioral results showed that participants were significantly slower in reading words in their less proficient language (Spanish) than in their more proficient language (English). fMRI results also revealed that reading words in the less proficient language yielded greater activity in the articulatory motor system, consisting of supplementary motor area/cingulate, insula, and putamen. Together, the behavioral and fMRI results suggest that the less practiced, hence less proficient, language requires greater articulatory motor effort, which results in slower reading rates. Moreover, we found that orthographic transparency also played a neuromodulatory role. More transparent Spanish words yielded greater activity in superior temporal gyrus (STG; BA 22), a region implicated in phonological processing, and orthographically opaque English words yielded greater activity in visual processing and word recoding regions, such as the occipito-parietal border and inferior parietal lobe (IPL; BA 40). Overall, our fMRI results suggest that the articulatory motor system is more plastic, hence, more amenable to change because of greater exposure to the L2. By contrast, we propose that our orthography effect is less plastic, hence, less influenced by frequency of exposure to a language system. PMID- 16242352 TI - Osteoarthritis cartilage histopathology: grading and staging. AB - OBJECTIVE: Current osteoarthritis (OA) histopathology assessment methods have difficulties in their utility for early disease, as well as their reproducibility and validity. Our objective was to devise a more useful method to assess OA histopathology that would have wide application for clinical and experimental OA assessment and would become recognized as the standard method. DESIGN: An OARSI Working Group deliberated on principles, standards and features for an OA cartilage pathology assessment system. Using current knowledge of the pathophysiology of OA morphologic features, a proposed system was presented at OARSI 2000. Subsequently, this was widely circulated for comments amongst experts in OA pathology. RESULTS: An OA cartilage pathology assessment system based on six grades, which reflect depth of the lesion and four stages reflecting extent of OA over the joint surface was developed. CONCLUSIONS: The OARSI cartilage OA histopathology grading system appears consistent and simple to apply. Further studies are required to confirm the system's utility. PMID- 16242354 TI - A history of the understanding of cartilage. AB - OBJECTIVE: To review the historic development of the understanding of articular cartilage from the earliest comment in the fourth century BCE until about 2000. DESIGN: The history up to 1900 is told chronologically, divided into (1) recognition of the tissue, (2) structure, and (3) chemistry. The twentieth century is sketched with a timeline of discoveries that at the time were important and a bibliography of journal review articles. RESULTS: By 1900 the avascular, aneural state and fibrillar composition have been accepted. The nutrition of articular cartilage remained in dispute. The composition of the binding substance and its relation to collagen remained unknown. Research in the first half of the twentieth century continued to be impeded by lack of technology. The advent of electron microscopy, isotopic tracer technics and enzymology rapidly accelerated the understanding of hyaline cartilage beginning in the 1950s. CONCLUSIONS: The history of research on hyaline cartilage illustrates the dependence of scientific progress on technologic innovation. PMID- 16242353 TI - Intra-articular hyaluronic acid compared with corticoid injections for the treatment of rhizarthrosis. AB - OBJECTIVE: Prospective assessment of the efficacy and tolerability of intra articular sodium hyaluronate (SH; Ostenil mini) and triamcinolone acetonide (TA; Volon A10) for treatment of osteoarthritis (OA) of the carpometacarpal (CMC) joint of the thumb in a 26-week, controlled, randomized, on an intention to treat, masked-observer study. METHODS: Patients were treated with three intra articular injections of either SH (n = 28) or TA (n = 28). Primary assessments were pain according to a 100mm visual analogue scale and extensive clinical and functional parameters such as swelling, grip power and range of motion. The population was analysed using one- and two-sided Mann-Whitney (MW) estimators. RESULTS: Maximum pain relief occurred at 2-3 weeks for TA and at week 26 for SH after the first intra-articular injection. At weeks 2-3 TA was significantly better than SH (MW: 0.3319 and 0.3063; P = 0.9827 and 0.9929). At week 26 a slight superiority of SH could be observed (MW: 0.53; P = 0.3624) and non inferiority could be proven. After 26 weeks lateral pinch power was significantly better in the SH-group (MW: 0.6331; P = 0.0226). In all, 88.0% of patients treated with SH and 79.1% of the TA-group described pain improvement after 26 weeks. Both agents were well tolerated. No adverse events with causal connection to the investigational products occurred. CONCLUSION: A single course of three SH injections is effective in relieving pain and improving joint function in patients with OA of the CMC joint of the thumb. Although in comparison with triamcinolone its effects are achieved more slowly, the results indicate a superior long-lasting effect of hyaluronan at 6 months after end of treatment period. PMID- 16242355 TI - Autologous chondrocyte implantation: a systematic review. AB - OBJECTIVE: To critically analyze the existing literature relating to autologous chondrocyte implantation (ACI) and thereby to ascertain whether the technique is clinically effective and safe. METHODS: Using predefined criteria, we searched a number of automated databases, such as MEDLINE, EMBASE, Cochrane, CRD, etc., for relevant articles, which were then analyzed by two independent reviewers. RESULTS: Three clinical trials and nine case series were evaluated. The clinical trials yielded no evidence that ACI was superior to the therapeutic alternatives with which it was compared. In contrast, the case series revealed an improvement in patients. However, as with the clinical trials, the follow-up periods were usually very short. In general, few adverse effects were observed, indicating that ACI is a safe technique. CONCLUSION: Available data afford no evidence that ACI is more effective than other conventional techniques in treating chondral lesions of the knee. PMID- 16242356 TI - Cell columns in articular cartilage physes questioned: a review. AB - OBJECTIVE: To report the outcome of a search of the literature for evidence in support of the published [Brown RA, Blunn GW, Salisbury JR, Byers PD. Two patterns of calcification in primary (physeal) and secondary (epiphyseal) growth cartilage. Clin Orthop1993;294:318-24] observation that there are differences in the cellular organisation and ossification between the articular growth cartilage and the metaphyseal physis, i.e., dual patterns of ossification. METHOD: The search of the journal literature was by Medline. Many references came from found articles, and from textbooks. The source texts were at the libraries of the Royal Society of Medicine, The Wellcome Trust and the British Library. RESULTS: (1) The search produced nine authors whose observations make up the bulk of the paper, which support the dual pattern of physes. (2) But there were also articles in favour of the single pattern of cellular organisation and ossification, in which, nevertheless, there were illustrations which were inconsistent with this, and favoured the duality. (3) The third section of the results mainly concerns the role of osteoclasts in the ossification process in articular physes. They are generally regarded as insufficient in number to play a solo part. Quantitative data about osteoclasts are limited; mathematical modelling is proposed as a more objective test. CONCLUSION: Objective, assessable criteria are presented in favour of a modification in our understanding of articular cartilage, and could, and should be augmented by further testing of the hypothesis. Some lines of enquiry are suggested. PMID- 16242357 TI - Repeated ultrasound-guided intra-articular injections of 40 mg of Hyalgan may be useful in symptomatic relief of hip osteoarthritis. PMID- 16242358 TI - A low temperature method of isolating normal human articular chondrocytes. AB - OBJECTIVE: Numerous methods for isolation of human chondrocytes are reported in the literature, most based on isolation from animal cartilage. Normal human articular cartilage (NHAC) poses particular problems for isolating chondrocytes when compared to animal or other types of human cartilage: a hardy matrix, combined with few and friable chondrocytes makes isolation difficult. Our objective was to develop an efficient method of isolating chondrocytes from NHAC without jeopardising the viability of these cells. DESIGN: In this study we demonstrate that lowering the enzymatic digestion temperature to 27 degrees C increases cell yield and chondrocyte viability. We then optimised this low temperature isolation of chondrocytes from NHAC by comparing the relative efficacies of trypsin and protease and hyaluronidase in combination with different types of collagenase (I, II and XI) at releasing chondrocytes from their surrounding cartilaginous matrix. Enzymes were tested at different concentrations and for differing times. Outcome measures included determining the amount of cartilage digested, the number of viable chondrocytes isolated per gram of cartilage and cell adherence rates. CONCLUSIONS: From these set of experiments, the method that maximised cell yield without jeopardising cell viability proved to be a two stage process: pre-digestion step using trypsin for 15 min; followed by overnight digestion with a combination of two types of collagenase (types I and II) and at a lower temperature of 27 degrees C. This has resulted in an efficient and robust method of releasing chondrocytes from cartilage, without jeopardising the viability of these cells. PMID- 16242359 TI - Subchondral bone trauma causes cartilage matrix degeneration: an immunohistochemical analysis in a canine model. AB - Joint instability was believed to be the main cause of osteoarthritis following non-fracture articular trauma. However, sudden high impact load through articular cartilage onto subchondral bone may also cause osteoarthritic changes. OBJECTIVE: We asked whether early osteoarthritic changes following transarticular impact may be depicted using immunofluorescence on unfixed cryosections to contribute to a more detailed understanding of degenerative processes of joint impaction. DESIGN: Transarticular impacts were applied to patellofemoral joints of 12 skeletally mature beagle dogs (age: 15-16 months) using a drop tower. Biopsies of impact areas were sampled after 6 months and processed for standard light microscopy on formalin-fixed sections and for immunofluorescence for collagen type I (col I), type II (col II) and aggrecan (AC) on unfixed cryosections. Gross morphology and immunofluorescence on cryosections were documented using a semi-quantitative scaling system, compared to healthy controls and to standard light microscopy. RESULTS: Four biopsies showed almost entirely fibrocartilaginous morphology, four appeared to be of preserved hyaline morphology with only minor signs of fibrocartilaginous remodelling and four showed preserved hyaline appearance. We found decrease in col II and AC expression in highly degenerative specimens as well as increase of col I expression. Increased col I expression in the pericellular matrix could even be depicted in specimens with intact hyaline morphology. DISCUSSION/CONCLUSION: Observations suggest that joint impaction causes early osteoarthritic changes after 6 months. Collagen network disruption seems to lead to AC loss, although other researchers found isolated AC loss without denaturation of col II using immunofluorescence in formalin-fixed specimens. This is the first study on effects of transarticular impact using immunofluorescence on unfixed cryosections. PMID- 16242360 TI - Magnetic resonance imaging characterization of osteochondral defect repair in a goat model at 8 T. AB - OBJECTIVE: This study was performed to non-invasively visualize and characterize osteochondral (OC) repair in ex vivo goat stifles using an 8 T magnetic resonance imaging (MRI) scanner and to compare the MR morphology with images obtained from 1.5 T, gross morphology and histology. METHODS: Mature, neutered male goats were assigned to an 8-week (n = 4) or 16-week (n = 4) study period. Two cylindrical OC defects (7 mm diameter, full cartilage thickness and 1mm into subchondral bone) were surgically created in the right stifle: one in the medial femoral condyle (MFC) and the other in the trochlear groove (TG). The implant matrices (non-woven or foam) were secured in the defect using a bottom anchored fixation device (FD). The contralateral left stifles served as time zero controls. At the day of necropsy, implants were placed at both defect sites (MFC and TG) on the normal left stifles. Following necropsy, the ex vivo goat stifles (intact and encapsulated) were disarticulated. Within 24 h postnecropsy, MR scans of the stifles along the mid-sagittal plane of the OC defect were acquired at 8 T and 1.5 T. MR relaxation times, T1 and T2, were measured at the region of repair tissue (RT) and adjacent native cartilage. Immediately after MR imaging, the stifles were dissected, grossly examined, and a sagittal OC block corresponding to the MR region of interest was prepared for formalin fixation. RESULTS: The high-resolution MR images enabled visualization of cartilage and bone integrity surrounding the implant as well as delineating the margins of RT/implant matrix and the FD. On spin echo sequence, the RT variably appeared as high, intermediate or low MR signal intensity; whereas, the FD always appeared as low signal intensity. In general, the MR signal intensity of 8-week RT was slightly higher compared to 16-week RT; however, there was no difference in RT morphology of stifles implanted with the non-woven matrix or foam matrix. Subchondral sclerosis appeared as low signal intensity. The 8 T MR images showed better delineation of the stifle tissues compared to the images acquired at 1.5 T. The T2 relaxation time of the RT appears to indicate (inconclusive due to small number of samples) a slight variation in the RT type between 8 weeks and 16 weeks. At both study times, the defects grossly appeared whitish to reddish but did not have the characteristic hyaline appearance typical of articular cartilage (AC). The gross appearance of the MFC and TG RT differed, which was predominantly mottled and recessed with fissuring of adjacent native AC in the MFC. Histologically, the RT of both 8-week and 16-week postsurgical defects predominantly comprised fibrovascular connective tissue with only few samples showing the presence of fibrocartilaginous and/or hypertrophic chondrocytes within the defect RT at 8 weeks. Also, compared to 8-week, the 16-week RT appeared to be more fibrotic. CONCLUSION: Using 8 T scanner, high-resolution MR images of ex vivo encapsulated goat stifles confirmed the capability of high-field MR imaging to distinguish the defect RT from the FD and adjacent joint tissues. The extent of OC repair and adjacent bone lesions (at 8 weeks and 16 weeks) observed in the MR images compared well with those observed on the corresponding histological sections. PMID- 16242361 TI - A double-blind randomized controlled trial comparing alternate forms of high molecular weight hyaluronan for the treatment of osteoarthritis of the knee. AB - OBJECTIVE: To compare the safety and effectiveness of a high molecular weight hyaluronan produced by biological fermentation (Bio-HA) with those of avian derived hyaluronan that uses cross-linking to achieve high molecular weight (CL HA). DESIGN: This was a prospective, multicenter, randomized, double-blind trial evaluating patients with confirmed osteoarthritis of the knee. The Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC Index) pain subscale was the primary effectiveness measure (visual analog scale). Both products were administered via three weekly injections, with follow-up evaluations at weeks 3, 6 and 12. Acetaminophen was permitted as rescue medication and quantitated by pill counts. RESULTS: Analyses were performed on the intent-to-treat population, defined as all patients receiving at least one injection. Of the 321 patients randomized to treatment, 314 patients (98%) completed the final study assessment. Improvement in the average WOMAC Index pain score was 29.8mm (-61.6%) for Bio-HA and 28.8mm (-54.9%) for CL-HA, meeting the prospective criteria for non-inferiority. For the secondary outcome measures, statistically significant differences favored Bio-HA for the number of patients requiring acetaminophen (P=0.013) and patient global satisfaction evaluations (P=0.03). Local reactions differed between the products in that 15 effusions were reported in 13 CL-HA patients (8.1%) after injection, compared to one effusion (0.6%) after Bio-HA injection (P=0.0015). CONCLUSION: The effectiveness of Bio-HA was not inferior to that of CL-HA. The significantly higher incidence of post injection effusion in the CL-HA group provides a safety advantage for Bio-HA. These data suggest that Bio-HA has an improved benefit-risk profile compared with CL-HA. PMID- 16242362 TI - Osteoarthritis cartilage histopathology: grading and staging. PMID- 16242363 TI - Mesogleal cells of the jellyfish Aurelia aurita are involved in the formation of mesogleal fibres. AB - The extracellular matrix of the jellyfish Aurelia aurita (Scyphozoa, Cnidaria), known as the mesoglea, is populated by numerous mesogleal cells (Mc). We determined the pattern of the Mc and the mesoglea, raised polyclonal antibodies (RA47) against the major mesogleal protein pA47 (47 kDa) and checked their specificity. In the mesoglea, RA47 stains pA47 itself. In immunoblots of Mc, RA47 stains bands of 120 kDa and 80 kDa; weaker staining is observed at pA47. The same staining pattern is seen on blots of jellyfish epidermal cells and of whole Hydra (Hydrozoa) or isolated mesoglea of Hydra. Our data indicate that pA47 is synthesized by Mc and epidermal cells as high molecular precursors. Using immunostaining techniques, we showed Mc to be involved in the formation of mesogleal non-collagenous (called "elastic" in classic morphological studies) fibres. The biochemical and morphological data suggest that Mc originate from the epidermis. PMID- 16242364 TI - Sleep, breathing and the nose. AB - During sleep there is a discrete fall in minute ventilation and an associated increase in upper airway resistance. In normal subjects, the nasal part of the upper airway contributes only little to the elevation of the total resistance, which is mainly the consequence of pharyngeal narrowing. Yet, swelling of the nasal mucosa due to congestion of the submucosal capacitance vessels may significantly affect nasal airflow. In many healthy subjects an alternating pattern of congestion and decongestion of the nasal passages is observed. Some individuals demonstrate congestion of the ipsilateral half of the nasal cavity when lying down on the side. Nasal diseases, including structural anomalies and various forms of rhinitis, tend to increase nasal resistance, which typically impairs breathing via the nasal route in recumbency and during sleep. A role of nasal obstruction in the pathogenesis of sleep-disordered breathing has been implicated by many authors. While it proves difficult to show a relationship between the degree of nasal obstruction and the number of disturbed breathing events, the presence of nasal obstruction will most likely have an impact on the severity of sleep-disordered breathing. Identification of nasal obstruction is important in the diagnostic work-up of patients suffering from snoring and sleep apnea. PMID- 16242365 TI - Use of sodium nitroprusside in the anaesthetic protocol of a patent ductus arteriosus ligation in a dog. AB - Patent ductus arteriosus (PDA) is one of the most common congenital cardiac abnormalities in the dog. Sodium nitroprusside has been proposed as an infusion during PDA ligation as the resultant nitric oxide production causes hypotension thereby facilitating surgical dissection and decreasing the risk of haemorrhage. This case report details how the method described did not decrease blood pressure and therefore had no beneficial effects. It is speculated that the lack of response may have been due to tachyphylaxis. Other drugs may be preferred to induce deliberate hypotension in the dog. PMID- 16242366 TI - Effects of temperature and feed intake on astaxanthin digestibility and metabolism in Atlantic salmon, Salmo salar. AB - The effects of feed intake, growth rate and temperature (8 and 12 degrees C) on apparent digestibility coefficients (ADC), blood uptake of individual astaxanthin E/Z isomers and metabolism of astaxanthin (3,3'-dihydroxy-beta,beta-carotene-4,4' dione) were determined in Atlantic salmon. Accumulation of idoxanthin (3,4,3' trihydroxy-beta,beta-carotene-4-one) in plasma was used to indicate metabolic transformation of astaxanthin. Quadruplicate groups of fish were subjected to three different treatments; one treatment was kept at 12 degrees C and fed to satiation. Another treatment kept at 12 degrees C was pair-fed with fish fed to satiation at 8 degrees C, resulting in a restricted feeding regime for the former treatment. After 2 months of feeding, the fish were fed a single meal containing ballotini glass beads to determine individual feed intake and Y(2)O(3) as an inert marker to determine ADCs. The faeces samples were pooled into 6 categories according to individual meal size (range 0.2-1.5% of body weight) and the ADCs for different meal sizes were determined. ADCs of astaxanthin ranged from 20% to 60% but were not significantly correlated with meal size. However, fish kept at 12 degrees C had approximately 10% higher ADC than fish kept at 8 degrees C (p=0.032). Growth rate and plasma astaxanthin concentration were higher at higher temperature and higher ration. Plasma concentration of idoxanthin was not affected by temperature or by meal size. The incidence of fin erosion and non feeding individuals was significantly higher among fish fed a restricted ration indicating more aggressive interactions. Fish with visible fin damage had a tendency for having higher idoxanthin content in plasma than fish without noticeable fin damage. It is concluded that temperature but not individual meal size affect ADC of astaxanthin, whereas both influence plasma astaxanthin levels and may therefore affect the efficiency of astaxanthin utilization. PMID- 16242367 TI - Total lactate dehydrogenase activity of tail muscle is not cold-adapted in nocturnal lizards from cool-temperate habitats. AB - The dependence of metabolic processes on temperature constrains the behavior, physiology and ecology of many ectothermic animals. The evolution of nocturnality in lizards, especially in temperate regions, requires adaptations for activity at low temperatures when optimal body temperatures are unlikely to be obtained. We examined whether nocturnal lizards have cold-adapted lactate dehydrogenase (LDH). LDH was chosen as a representative metabolic enzyme. We measured LDH activity of tail muscle in six lizard species (n=123: three nocturnal, two diurnal and one crepuscular) between 5 and 35 degrees C and found no differences in LDH-specific activity or thermal sensitivity among the species. Similarly, the specific activity and thermal sensitivity of LDH were similar between skinks and geckos. Similar enzyme activities among nocturnal and diurnal lizards indicate that there is no selection of temperature specific LDH enzyme activity at any temperature. As many nocturnal lizards actively thermoregulate during the day, LDH may be adapted for a broad range of temperatures rather than adapted specifically for the low temperatures encountered when the animals are active. The total activity of LDH in tropical and temperate lizards is not cold-adapted. More data are required on biochemical adaptations and whole animal thermal preferences before trends can be established. PMID- 16242368 TI - Histoplasmosis and penicilliosis: comparison of clinical features, laboratory findings and outcome. AB - INTRODUCTION: Histoplasmosis and penicilliosis are infections caused by the dimorphic fungi, Histoplasma capsulatum and Penicillium marneffei, respectively. The aim of this study was to compare the clinical presentation, laboratory and radiologic findings and outcome of these infections at Srinagarind Hospital, Khon Kaen, Thailand. METHODS: The medical records of patients who had positive cultures for Histoplasma capsulatum and Penicillium marneffei during 1996-2002 were reviewed. The data were compared and analyzed by the Chi-square and Fisher's exact tests. RESULTS: There were 32 and 36 medical records of patients with H. capsulatum and P. marneffei infection, respectively, available for review. The most common underlying disease of patients with histoplasmosis and penicilliosis was acquired immunodeficiency syndrome (AIDS), which accounted for 90.6% and 91.7%, respectively. The most common clinical findings in both infections were fever, weight loss, cough, anemia, lymphadenopathy, hepatomegaly and splenomegaly. Frequencies of skin lesions were not statistically different between either group (P=0.20). Laboratory findings were similar between the two infections, except hyperbilirubinemia, which was more common in the penicilliosis group (P=0.02). There were similar abnormal X-ray findings in both groups with interstitial infiltration the most common abnormality. CONCLUSIONS: Histoplasmosis and penicilliosis had similar clinical presentations, laboratory findings and chest X-ray abnormalities. Itraconazole is recommended as secondary prophylaxis in HIV-infected patients who have histoplasmosis or penicilliosis. PMID- 16242369 TI - [An exchange without negotiation: medical screening of blood donors in France]. AB - In France, screening of blood donors is realized through an interview with a medical doctor. This article offers a sociological analysis of these interviews in an interactionist perspective. 80 interviews have been observed and analysed. Two types of situations are distinguished. In most cases, no problem appears in the interview so that the possibility of the gift is not call into question, whereas this is not the case in a small part of the corpus, in which the doctor starts a thorough discussion with the donor. From this description, the author analyzes the logics of the transfusion: the logic of the donors, who want to give their blood, opposes to that of the doctors, who seek to identify the possible risks. The article highlights the "pact" which binds the donors and the transfusion service. The donors accept the absolute power of the doctors provided they are able to give their blood. Contrary to the usual relations between doctors and patients, this situation excludes any negotiation. PMID- 16242371 TI - Infection-induced respiratory burst in BALB/c macrophages kills Leishmania guyanensis amastigotes through apoptosis: possible involvement in resistance to cutaneous leishmaniasis. AB - The immune mechanisms that underlie resistance and susceptibility to leishmaniasis are not completely understood for all species of Leishmania. It is becoming clear that the immune response, the parasite elimination by the host and, as a result, the outcome of the disease depend both on the host and on the species of the infecting Leishmania. Here, we analyzed the outcome of the infection of BALB/c mice with L. guyanensis in vivo and in vitro. We showed that BALB/c mice, which are a prototype of susceptible host for most species of Leishmania, dying from these infections, develop insignificant or no cutaneous lesions and eliminate the parasite when infected with promastigotes of L. guyanensis. In vitro, we found that thioglycollate-elicited BALB/c peritoneal macrophages, which are unable to eliminate L. amazonensis without previous activation with cytokines or lipopolysaccharide, can kill L. guyanensis amastigotes. This is the first report showing that infection of peritoneal macrophages with stationary phase promastigotes efficiently triggers innate microbicidal mechanisms that are effective in eliminating the amastigotes, without exogenous activation. We demonstrated that L. guyanensis amastigotes die inside the macrophages through an apoptotic process that is independent of nitric oxide and is mediated by reactive oxygen intermediates generated in the host cell during infection. This innate killing mechanism of macrophages may account for the resistance of BALB/c mice to infection by L. guyanensis. PMID- 16242372 TI - Melatonin and its kynurenin-like oxidation products affect the microbicidal activity of neutrophils. AB - Activated phagocytes oxidize the hormone melatonin to N1-acethyl-N2-formyl-5 methoxykynuramine (AFMK) in a superoxide anion- and myeloperoxidase-dependent reaction. We examined the effect of melatonin, AFMK and its deformylated-product N-acetyl-5-methoxykynuramine (AMK) on the phagocytosis, the microbicidal activity and the production of hypochlorous acid by neutrophils. Neither neutrophil and bacteria viability nor phagocytosis were affected by melatonin, AFMK or AMK. However these compounds affected the killing of Staphylococcus aureus. After 60 min of incubation, the percentage of viable bacteria inside the neutrophil increased to 76% in the presence of 1 mM of melatonin, 34% in the presence of AFMK and 73% in the presence of AMK. The sole inhibition of HOCl formation, expected in the presence of myeloperoxidase substrates, was not sufficient to explain the inhibition of the killing activity. Melatonin caused an almost complete inhibition of HOCl formation at concentrations of up to 0.05 mM. Although less effective, AMK also inhibited the formation of HOCl. However, AFMK had no effect on the production of HOCl. These findings corroborate the present view that the killing activity of neutrophils is a complex phenomenon, which involves more than just the production of reactive oxygen species. Furthermore, the action of melatonin and its oxidation products include additional activities beyond their antioxidant property. The impairment of the neutrophils' microbicidal activity caused by melatonin and its oxidation products may have important clinical implications, especially in those cases in which melatonin is pharmacologically administered in patients with infections. PMID- 16242373 TI - Bone loss associated with anorexia nervosa. AB - The objective of this study was to evaluate the epidemiology, diagnosis, pathophysiology, and treatment of bone loss related to anorexia nervosa. Earlier onset and longer duration of anorexia nervosa are associated with more severe bone loss. Osteoporosis develops in 38-50% of cases. Bone mineral density measurement by dual-energy X-ray absorptiometry is useful for assessing bone mass, and bone marker assays provide information on bone turnover. Bone loss in anorexia nervosa is probably multifactorial. Estrogen deficiency was long felt to be the major factor. However, in contrast to postmenopausal osteoporosis, bone loss associated with anorexia nervosa is related mainly to inadequate bone formation, with only a slight increase in bone resorption. This suggests a role for nutritional factors, such as disturbances in the growth hormone-somatomedin C axis (GH/IGF-I) related to malnutrition. The best treatment strategy for correcting bone mass in patients with anorexia nervosa is not agreed on. Resumption of menstrual cycles and weight gain seem necessary but not always sufficient. Studies found no benefits with estrogen therapy, but this was usually given as estrogen-progestin contraceptives. No vast studies evaluating hormone replacement therapy have been reported. Bone formation enhancers such as IGF-I seem to provide the best results, most notably when used in combination with estrogens. This suggests that complex treatment strategies combining bone formation enhancers and bone resorption inhibitors may deserve evaluation. PMID- 16242370 TI - Toll-like receptor 2-mediated expression of beta-defensin-2 in human corneal epithelial cells. AB - We previously showed that human corneal epithelial cells (HCECs) express Toll like receptors (TLRs), which recognize gram-positive bacteria and respond to Staphylococcus aureus infection by the expression and secretion of proinflammatory cytokines and beta-defensin-2 (hBD2). In this study, we further elucidated the underlying mechanisms regulating hBD-2 expression and its role in innate defense in HCECs in response to S. aureus challenge. Exposure of HUCL cells, a telomerase-immortalized HCEC line, to S. aureus, its exoproducts (1:10 dilution), or synthetic lipopeptide Pam3Cys (10 microg/ml) resulted in the up regulation of hBD-2, but not hBD1 and hBD3. Similar to HUCL cells, primary HCECs responded to S. aureus-exoproducts and Pam3Cys challenge by expressing hBD2 mRNA and secreting hBD2 into the culture media. Furthermore, these stimuli induced the expression of TLR2 at both mRNA and protein levels. Consistently with its role as a major pattern-recognizing receptor, TLR2 was located at the cell surface by cell surface biotinylation. The treatment of HUCL cells with TLR2 neutralizing antibody resulted in a significant decrease in Pam3Cys-induced hBD2 production as well as IL-6, IL-8, and TNF-alpha secretion. The Pam3Cys-induced hBD2 expression was completely blocked by NF-kappaB inhibitors and partially inhibited by p38 MAP kinase and the JNK inhibitors. Conditioned media derived from HCECs challenged with S. aureus-exoproducts or Pam3Cys exhibited antibacterial activity against S. aureus, Pseudomonas aeruginosa and Escherichia coli. These findings suggest that S. aureus induces hBD2 production through TLR2-mediated pathways in HCECs and that pathogen-challenged, TLR-activated HCECs possess antimicrobial activity. Thus, the epithelium might play a role in innate defense against bacterial infection by directly killing bacteria in the cornea. PMID- 16242374 TI - Contribution of helical computed tomography to the evaluation of early hip osteoarthritis: a study in 18 patients. AB - OBJECTIVES: To show that helical computed tomography arthrograms (HCTA) with multiplanar reformations can document cartilage lesions and their characteristics in patients with suspected hip osteoarthritis and normal or inconclusive hip radiographs. METHODS: We retrospectively reviewed abnormal HCTAs from patients with mechanical hip pain and normal findings on anteroposterior and oblique ("faux profil") radiographs of the pelvis. HCTA in all patients consisted in acquiring 1 mm transverse slices and obtaining coronal and sagittal reformations. Radiographs and HCTAs were read separately by two experienced radiologists, who then worked together to reach a consensus. RESULTS: We identified 18 patients with abnormal HCTA findings and a full set of imaging studies. Mean age was 47.8 years, and there were 14 women and four men. Acetabular cartilage lesions were found consistently; they were often deep and predominated in the anterosuperior region. A labral fissure was noted in 12 patients. The sagittal and coronal reformations proved more informative than the acquired transverse slices. Interobserver reproducibility was excellent for the HCTA diagnosis of cartilage lesions. CONCLUSIONS: In patients with hip pain and normal radiographs, HCTA can provide a diagnosis of hip osteoarthritis by showing cartilage lesions, which are usually located in the anterosuperior part of the acetabulum. PMID- 16242375 TI - Ultrasonic versus silent methylation of vegetable oils. AB - The profile of fatty acid methyl esters (FAME) of different vegetable oils produced under ultrasonic irradiation and conventional heating were compared. In the presence of potassium hydroxide as catalyst, the distribution of FAME was quite similar for both procedures, while in the case of sodium hydroxide ultrasonic irradiation gave better results. The FAME profile resulted from the reaction catalyzed by sulfuric acid was almost the same with the one resulted from the reaction catalyzed by KOH, while boron trifluoride can give rise to many artifacts, thus is not a reliable catalyst. PMID- 16242376 TI - The lumbar multifidus muscle and patterns of pain. AB - This paper describes the patterns of pain induced by injecting hypertonic saline into the lumbar multifidus muscle opposite the L5 spinous process in 15 healthy adult volunteers. All subjects experienced local pain while referred pain was reported by 13 subjects in one of two regions of the thigh; anterior (n=5) or posterior (n=8). These results confirm that the multifidus muscle may be a source of local and referred pain. Comparison of these maps with pain maps following stimulation of the L4 medial dorsal rami and L4-5 interspinous ligaments shows that pain arising from the band of multifidus innervated by the L4 dorsal ramus has a segmental distribution. In addition patterns of pain arising from multifidus clearly overlap those reported for other lumbar structures. These findings highlight the difficulty of using pain distribution to accurately identify specific lumbar structures as the source of pain. PMID- 16242377 TI - Dipeptidyl peptidase IV inhibitors: a promising new therapeutic approach for the management of type 2 diabetes. AB - Glucagon-like peptide-1 is an insulinotropic hormone with antidiabetic potential due to its spectrum of effects, which include glucose-dependent stimulation of insulin and inhibition of glucagon secretion, tropic effects on the pancreatic beta-cells, inhibition of gastric emptying and the reduction of appetite. Glucagon-like peptide-1 is, however, extremely rapidly inactivated by the serine peptidase, dipeptidyl peptidase IV, so that the native peptide is not useful clinically. A new approach to utilise the beneficial effects of glucagon-like peptide-1 in the treatment of type 2 diabetes has been the development of orally active dipeptidyl peptidase IV inhibitors. Preclinical studies have demonstrated that this approach is effective in enhancing endogenous levels of glucagon-like peptide-1, resulting in improved glucose tolerance in glucose-intolerant and diabetic animal models. In recent studies of 3-12 months duration in patients with type 2 diabetes, dipeptidyl peptidase IV inhibitors have proved efficacious, both as monotherapy and when given in combination with metformin. Fasting and postprandial glucose concentrations were reduced, leading to reductions in glycosylated haemoglobin levels, while beta-cell function was preserved. Current information suggests dipeptidyl peptidase IV inhibitors are body weight neutral and are well tolerated. A number of dipeptidyl peptidase IV inhibitors are now in the late stages of clinical development. These have different properties, in terms of their duration of action and anticipated dosing frequency, but data from protracted dosing studies is presently not available to allow comparison of their clinical efficacy. PMID- 16242378 TI - Chemical approaches to deciphering the glycosaminoglycan code. AB - Glycosaminoglycans are sulfated biopolymers with rich chemical diversity and complex functions in vivo, contributing to processes ranging from cell growth and neuronal development to viral invasion and neurodegenerative disease. Recent studies suggest that glycosaminoglycans may encode information in the form of a 'sulfation code,' whereby discrete modifications to the polysaccharide backbone may direct the location or activities of proteins. PMID- 16242379 TI - Protein surface recognition and proteomimetics: mimics of protein surface structure and function. AB - Due to their key roles in a number of biological processes, protein-protein interactions are attractive and important targets, typically involving areas greater than 6 nm2. The disruption of such interactions remains a challenging feat but, in recent years, there has been considerable progress in the design of proteomimetics: molecules that mimic the structure and function of extended regions of protein surfaces. In particular, porphyrins, calixarenes, alpha helical mimetics and small molecules have successfully modulated significant protein-protein interactions, including those involved in cancer and HIV. PMID- 16242380 TI - Co-mutagenic activity of arsenic and benzo[a]pyrene in mouse skin. AB - Exposure to inorganic arsenic in drinking water is linked to skin, lung and bladder cancer in humans. The mechanism of arsenic-induced cancer is not clear, but exposure to arsenic and polycyclic arylhydrocarbons (PAH) is more carcinogenic than exposure to either type of carcinogen alone. Arsenic can also generate reactive oxygen species, suggesting that oxidation of DNA may play a role in carcinogenesis. Oxidization of guanosines in polyG tracts is known to cause frameshift mutations, and such events can be detected in situ using the G11 placental alkaline phosphatase (PLAP) transgenic mouse model, which reports frameshift mutations in a run of 11 G:C basepairs by generating cells containing heat-resistant alkaline phosphatase activity. PAH can also induce frameshift mutations. In the study described here, FVB/N mice carrying the G11 PLAP transgene were crossed to C57Bl/6 mice. Half of the hybrid mice were given drinking water with sodium arsenite (10 mg/L) for 10 weeks. Half of the arsenic treated mice were also exposed to benzo[a]pyrene (BaP) by skin painting (500 nmol/week) for 8 weeks. Another group of mice was exposed to BaP but not arsenic. The effect on frameshift mutation was assessed by staining sections of skin tissue to detect cells with PLAP activity. Arsenic alone had no significant effect. On average, mice given BaP alone had approximately three times more PLAP positive (PLAP+) cells. By contrast, mice exposed to both arsenic and BaP exhibited 10-fold more PLAP+ cells in the skin, and these cells were often arranged in large clusters, suggesting derivation from stem cells. Whereas combined treatment produced more PLAP+ cells, stable BaP adduct levels and arsenic burdens were not higher in mice exposed to both agents compared to mice exposed to either one agent or the other. PMID- 16242381 TI - Fabrication of high quality microarrays. AB - Fabrication of DNA microarray demands that between ten (diagnostic microarrays) and many hundred thousands of probes (research or screening microarrays) are efficiently immobilised to a glass or plastic surface using a suitable chemistry. DNA microarray performance is measured by parameters like array geometry, spot density, spot characteristics (morphology, probe density and hybridised density), background, specificity and sensitivity. At least 13 factors affect these parameters and factors affecting fabrication of microarrays are used in this review to compare different fabrication methods (spotted microarrays and in situ synthesis of microarrays) and immobilisation chemistries. PMID- 16242382 TI - The development of an optimized autologous blood donation program. AB - BACKGROUND: Autologous blood (ABD) programs have been in place in Canada since the late 1980s. Activity is decreasing in many sites and utilization averages 50%. METHODS: In an effort to optimize our ABD program we introduced one time single donation red cell collection (Trima) with and without erythropoietin and compared the results to those with standard red cell collections. A pre-pilot study assessed feasibility. RESULTS: To date a total of 59 patients have entered the three arm Trima/epo pilot involving prostate surgery patients: their pre surgical Hg 158 was compared to 108 in normal autologous control patients. By decreasing the standard pre-donation autologous requirement of three units of red cells to a single Trima collection and at $300 per autologous unit, this represents a cost savings of $36,000 in our 59 patients. When applied to the total autologous program at our hospital (450 per year), this extrapolates to a minimum savings of over $250,000 when using combined collection approaches. 32/59 patients in this pilot study have received only autologous blood, again reducing the number of allogeneic units that would have been required. The need for only a single autologous donation cuts nursing time by 2/3 and also saves laboratory money, since only a single unit is tested for infectious diseases. CONCLUSION: When applied to an autologous blood program, the use of the Trima machine combined with erythropoietin in appropriate cases, results in a decrease in the number of units collected and a higher pre-operative hemoglobin. This program should see application in other sites. PMID- 16242383 TI - Models of epilepsy in the developing and adult brain: implications for neuroprotection. AB - Repeated seizures cause a sequence of molecular and cellular changes in both the developing and adult brain, which may lead to intractable epilepsy. This article reviews this sequence of neuronal alterations, with emphasis on the kindling model. At each step, the opportunity exists for strategic intervention to prevent or reduce the downstream consequences of epileptogenesis and seizure-induced adverse plasticity. The concept of seizure-induced brain damage must be expanded to include behavioral and cognitive deficits, as well as structural neuronal damage and increased predisposition to seizures. PMID- 16242384 TI - Mediated amperometric biosensors for lactic acid based on carbon paste electrodes modified with baker's yeast Saccharomyces cerevisiae. AB - Carbon paste electrodes modified with baker's yeast Saccharomyces cerevisiae (a source of flavocytochrome b(2)) were investigated as amperometric biosensors for lactic acid. Phenazine methosulphate was used as a mediator. The optimal operational conditions of the electrodes were: an operating potential 0.0 V, solution pH 7.2, concentration of phenazine methosulphate in solution 0.2 mM. A linear range in the dependence of the current responses on the concentration of lactic acid was up to 1 mM. The suitability of the electrodes for determination of lactic acid in milk and dairy products such as kefir and yoghurt was tested. The yeast cells in the paste remained viable at least for 1 month. PMID- 16242385 TI - Clinical and economic choices in the treatment of respiratory infections in cystic fibrosis: comparing hospital and home care. AB - BACKGROUND: A cost-effectiveness evaluation comparing home-based and hospital based treatment with intravenous antibiotics for respiratory exacerbations in adults with cystic fibrosis (CF) has not been previously undertaken. METHODS: The study was conducted in a UK adult CF centre from a health service perspective. Clinical outcome and resource use data were obtained from a retrospective one year study and combined with unit cost data in an incremental economic analysis. The primary outcome measure was percentage change in FEV(1); "effectiveness" was defined as maintenance of baseline average FEV(1) over the one-year study period. RESULTS: 116 patients received 454 courses of intravenous antibiotics. At the end of 1 year, there had been a mean percentage decline in FEV(1) compared with baseline average for home-treated patients but an improvement for hospital treated patients (Tukey's HSD mean difference 10.1%, 95% CI 2.9 to 17.2, p = 0.003). Treatment was deemed "effective" in more hospital (58.8%) than home (42.6%) patients. The cost of hospital treatment was higher than home treatment (mean difference 9,005 pounds, 95% CI 3,507 to 14,700, p<0.001). The mean ICER was 46,098 pounds (2.5th and 97.5th percentiles -374,044 and 362,472). CONCLUSIONS: Hospital treatment was more effective but more expensive than home treatment. Potential methods to improve outcome at home should be considered but these may have resource implications. PMID- 16242386 TI - LC-MS solvent composition monitoring and chromatography alignment using mobile phase tracer molecules. AB - In the field of proteomics, reproducible liquid chromatographic description of analytes is often a key element for the differentiation or identification of proteins or peptides for clinical or biological research projects. However, analyte identification by retention time can be problematic in proteomics where lack of standardization can result in significantly different chromatography for the same analytes analyzed on different machines. Here we present a novel method of monitoring the mobile phase gradient of LC-MS/MS analyses by monitoring the ion current signal intensities of tracer molecules dissolved in the mobile phase solvents. The tracers' ion current signal intensities chronicled gradient fluctuations, did not adversely affect the number or quality of CID-based sequence identifications, and had lower run-to-run variance when compared to retention time. PMID- 16242387 TI - Metabolism of isometheptene in human urine and analysis by gas chromatography mass spectrometry in doping control. AB - A study of the metabolism of isometheptene, an antispasmodic drug, in man and comparison with heptaminol metabolism, is presented in this paper. Isometheptene and two metabolites were detected in human urine after oral administration of a tablet containing isometheptene mucate. The urine level of the parent drug, which is excreted during the first 24 h, was determined using gas chromatography-mass spectrometry, after alkaline extraction with organic solvent. A minor metabolite of isometheptene was converted to heptaminol in vitro under the acidic hydrolysis conditions used for the screening procedure of stimulants and narcotics in doping control analysis. PMID- 16242388 TI - Editorial response to author review in publishing science. PMID- 16242390 TI - New developments in fetal heart scanning: three- and four-dimensional fetal echocardiography. AB - This article reviews the possibilities of three- and four-dimensional (3- and 4D) fetal echocardiography. A volume data set of a fetal heart can be acquired as a static volume, as a real-time 3D volume or as an offline 4D volume cine using spatial and temporal image correlation (STIC) software. STIC is explained and the potentials of this modality are emphasized. The display of a fetal heart volume data set demonstrates the cross-sections of interest, using the multiplanar mode or tomographic multislice imaging, and different volume rendering tools. The latter include: surface, minimum, inversion and glass body modes. This review highlights the potential of acquiring a digital volume data set of a heart cycle for later offline evaluation, either for an offline diagnosis, a second opinion (e.g. via Internet link) or for teaching fetal echocardiography to trainees and sonographers. PMID- 16242391 TI - Structural adaptations to bone loss in aging men and women. AB - INTRODUCTION: Bone apposition on the subperiosteal surface and bone loss from the endocortical surface during aging establish the external diameter, total cross sectional area (tCSA), cortical thickness (Ct.Th) and the distance the cortex is placed from the neutral axis of a long bone, all determinants of bone strength. We tested the hypothesis that sex-related differences in these processes produces a sexual dimorphism in tibial fragility. METHODS: The above traits were assessed in 688 women and 561 men (20-102 years old) using peripheral QCT. RESULTS: Total and medullary areas were greater in young adult men than young adult women. As age advanced, in men, tCSA area increased by 0.79 SD, and medullary area increased by 0.54 SD so that cortical area, cortical thickness and minimum and maximum moments of inertia (Imin and Imax) were similar at all ages. In women, tCSA increased by 0.2 SD, while medullary area increased by 2.6 SD so that cortical area and thickness and the moments of inertia diminished. Cortical apparent volumetric bone mineral density (vBMD) declined more in women (by 3.1 SD) than men (by 0.5 SD). In both sexes, the lower the cortical apparent vBMD, the higher the tCSA (women R2 = 0.13, men R2 = 0.16, both P < 0.0001), whereas the lower the Ct.Th, the lower the tCSA (women R2 = 0.30, men R2 = 0.32, both P < 0.0001). CONCLUSIONS: Bone loss reduces cortical thickness and increases intracortical porosity. These changes tend to be compensated for by periosteal apposition in both sexes but more greatly in men than in women, perhaps because this mechanism may be ineffective when cortical thinning is severe. PMID- 16242393 TI - PDF has found its receptor. AB - The neuropeptide pigment-dispersing factor (PDF) is a crucial component of the insect circadian clock, but the identity of its receptor has long remained a mystery. In this issue of Neuron, three papers (Hyun et al., Lear et al., and Mertens et al.) identify the PDF receptor. PMID- 16242394 TI - Glial (and neuronal) cells missing. AB - The Glial cells missing transcription factor is necessary and sufficient to induce glial-cell fates in the Drosophila embryonic nervous system. A study by Chotard et al. in this issue of Neuron reveals that this "master regulator" of glial cell fate specification is also required (gasp!) to generate neurons. PMID- 16242395 TI - One circuit, two kinds of timing. AB - In this issue of Neuron, Gabernet et al. report an elegant series of in vivo and in vitro experiments that dissect a disynaptic circuit dictating the dynamic transition of cortical spiking responses to whisker stimulation from coincidence detection to temporal integration. PMID- 16242396 TI - Recovery in the blink of an eye. AB - Sensory deprivation sheds light on cortical plasticity mechanisms, but recovery of lost brain function may bear the greatest clinical relevance. Ramoa and colleagues now find that binocular recovery from monocular occlusion can be extraordinarily rapid, independent of protein synthesis, and precise. Reactivation of latent connections may then reverse amblyopia. PMID- 16242397 TI - Attractor neural networks and spatial maps in hippocampus. AB - Attractor neural network theory has been proposed as a theory for long-term memory. Recent studies of hippocampal place cells, including a study by Leutgeb et al. in this issue of Neuron, address the potential role of attractor dynamics in the formation of hippocampal representations of spatial maps. PMID- 16242398 TI - Building excitatory and inhibitory synapses: balancing neuroligin partnerships. AB - Processing of neural information is thought to occur by integration of excitatory and inhibitory synaptic inputs. As such, precise control mechanisms must exist to maintain an appropriate balance between each synapse type. Recent findings indicate that neuroligins and their synaptic binding partners modulate the development of both excitatory and inhibitory synapses. Here we highlight these findings and discuss a mechanism potentially involved in controlling the balance between excitation and inhibition. PMID- 16242399 TI - Contributions of the amygdala to emotion processing: from animal models to human behavior. AB - Research on the neural systems underlying emotion in animal models over the past two decades has implicated the amygdala in fear and other emotional processes. This work stimulated interest in pursuing the brain mechanisms of emotion in humans. Here, we review research on the role of the amygdala in emotional processes in both animal models and humans. The review is not exhaustive, but it highlights five major research topics that illustrate parallel roles for the amygdala in humans and other animals, including implicit emotional learning and memory, emotional modulation of memory, emotional influences on attention and perception, emotion and social behavior, and emotion inhibition and regulation. PMID- 16242400 TI - Innovations in the imaging of brain functions using fluorescent proteins. AB - Fluorescence imaging has enabled us to decipher spatiotemporal information coded in complex tissues. Genetically encoded probes that enable fluorescence imaging of excitable cell activity have been constructed by fusing fluorescent proteins to functional proteins that are involved in physiological signaling. The probes are introduced into an intact organism and targeted to specific tissues, cell types, or subcellular compartments, thereby allowing specific signals to be extracted more efficiently than was previously possible. In this primer, I will describe how this approach has met neuroscientists' demands and desires. PMID- 16242401 TI - Neuroscience at Johns Hopkins. PMID- 16242402 TI - PDF receptor signaling in Drosophila contributes to both circadian and geotactic behaviors. AB - The neuropeptide Pigment-Dispersing Factor (PDF) is a principle transmitter regulating circadian locomotor rhythms in Drosophila. We have identified a Class II (secretin-related) G protein-coupled receptor (GPCR) that is specifically responsive to PDF and also to calcitonin-like peptides and to PACAP. In response to PDF, the PDF receptor (PDFR) elevates cAMP levels when expressed in HEK293 cells. As predicted by in vivo studies, cotransfection of Neurofibromatosis Factor 1 significantly improves coupling of PDFR to adenylate cyclase. pdfr mutant flies display increased circadian arrhythmicity, and also display altered geotaxis that is epistatic to that of pdf mutants. PDFR immunosignals are expressed by diverse neurons, but only by a small subset of circadian pacemakers. These data establish the first synapse within the Drosophila circadian neural circuit and underscore the importance of Class II peptide GPCR signaling in circadian neural systems. PMID- 16242403 TI - A G protein-coupled receptor, groom-of-PDF, is required for PDF neuron action in circadian behavior. AB - The neuropeptide Pigment-Dispersing Factor (PDF) plays a critical role in mediating circadian control of behavior in Drosophila. Here we identify mutants (groom-of-PDF; gop) that display phase-advanced evening activity and poor free running rhythmicity, phenocopying pdf mutants. In gop mutants, a spontaneous retrotransposon disrupts a coding exon of a G protein-coupled receptor, CG13758. Disruption of the receptor is accompanied by phase-advanced oscillations of the core clock protein PERIOD. Moreover, effects on circadian timing induced by perturbation of PDF neurons require gop. Yet PDF oscillations themselves remain robust in gop mutants, suggesting that GOP acts downstream of PDF. gop is expressed most strongly in the dorsal brain in regions that lie in proximity to PDF-containing nerve terminals. Taken together, these studies implicate GOP as a PDF receptor in Drosophila. PMID- 16242404 TI - A splice code for trans-synaptic cell adhesion mediated by binding of neuroligin 1 to alpha- and beta-neurexins. AB - Previous studies suggested that postsynaptic neuroligins form a trans-synaptic complex with presynaptic beta-neurexins, but not with presynaptic alpha neurexins. Unexpectedly, we now find that neuroligins also bind alpha-neurexins and that alpha- and beta-neurexin binding by neuroligin 1 is regulated by alternative splicing of neuroligin 1 (at splice site B) and of neurexins (at splice site 4). In neuroligin 1, splice site B is a master switch that determines alpha-neurexin binding but leaves beta-neurexin binding largely unaffected, whereas alternative splicing of neurexins modulates neuroligin binding. Moreover, neuroligin 1 splice variants with distinct neurexin binding properties differentially regulate synaptogenesis: neuroligin 1 that binds only beta neurexins potently stimulates synapse formation, whereas neuroligin 1 that binds to both alpha- and beta-neurexins more effectively promotes synapse expansion. These findings suggest that neuroligin binding to alpha- and beta-neurexins mediates trans-synaptic cell adhesion but has distinct effects on synapse formation, indicating that expression of different neuroligin and neurexin isoforms specifies a trans-synaptic signaling code. PMID- 16242405 TI - glial cells missing and gcm2 cell autonomously regulate both glial and neuronal development in the visual system of Drosophila. AB - The transcription factors Glial cells missing (Gcm) and Gcm2 are known to play a crucial role in promoting glial-cell differentiation during Drosophila embryogenesis. Our findings reveal a central function for gcm genes in regulating neuronal development in the postembryonic visual system. We demonstrate that Gcm and Gcm2 are expressed in both glial and neuronal precursors within the optic lobe. Removal of gcm and gcm2 function shows that the two genes act redundantly and are required for the formation of a subset of glial cells. They also cell autonomously control the differentiation and proliferation of specific neurons. We show that the transcriptional regulator Dachshund acts downstream of gcm genes and is required to make lamina precursor cells and lamina neurons competent for neuronal differentiation through regulation of epidermal growth factor receptor levels. Our findings further suggest that gcm genes regulate neurogenesis through collaboration with the Hedgehog-signaling pathway. PMID- 16242407 TI - Drosophila GPCR Han is a receptor for the circadian clock neuropeptide PDF. AB - The pigment-dispersing factor (PDF) is a neuropeptide controlling circadian behavioral rhythms in Drosophila, but its receptor is not yet known. From a large scale temperature preference behavior screen in Drosophila, we isolated a P insertion mutant that preferred different temperatures during the day and night. This mutation, which we named han, reduced the transcript level of CG13758. We found that Han was expressed specifically in 13 pairs of circadian clock neurons in the adult brain. han null flies showed arrhythmic circadian behavior in constant darkness. The behavioral characteristics of han null mutants were similar to those of pdf null mutants. We also found that PDF binds specifically to S2 cells expressing Han, which results in the elevation of cAMP synthesis. Therefore, we herein propose that Han is a PDF receptor regulating circadian behavioral rhythm through coordination of activities of clock neurons. PMID- 16242406 TI - Evidence that embryonic neurons regulate the onset of cortical gliogenesis via cardiotrophin-1. AB - Precursor cells of the embryonic cortex sequentially generate neurons and then glial cells, but the mechanisms regulating this neurogenic-to-gliogenic transition are unclear. Using cortical precursor cultures, which temporally mimic this in vivo differentiation pattern, we demonstrate that cortical neurons synthesize and secrete the neurotrophic cytokine cardiotrophin-1, which activates the gp130-JAK-STAT pathway and is essential for the timed genesis of astrocytes in vitro. Our data indicate that a similar phenomenon also occurs in vivo. In utero electroporation of neurotrophic cytokines in the environment of embryonic cortical precursors causes premature gliogenesis, while acute perturbation of gp130 in cortical precursors delays the normal timed appearance of astrocytes. Moreover, the neonatal cardiotrophin-1-/- cortex contains fewer astrocytes. Together, these results describe a neural feedback mechanism; newly born neurons produce cardiotrophin-1, which instructs multipotent cortical precursors to generate astrocytes, thereby ensuring that gliogenesis does not occur until neurogenesis is largely complete. PMID- 16242408 TI - Molecular mechanism of AMPA receptor noncompetitive antagonism. AB - AMPA-type glutamate receptors are specifically inhibited by the noncompetitive antagonists GYKI-53655 and CP-465,022, which act through sites and mechanisms that are not understood. Using receptor mutagenesis, we found that these antagonists bind at the interface between the S1 and S2 glutamate binding core and channel transmembrane domains, specifically interacting with S1-M1 and S2-M4 linkers, thereby disrupting the transduction of agonist binding into channel opening. We also found that the antagonists' affinity is higher for agonist unbound receptors than for activated nondesensitized receptors, further depending on the level of S1 and S2 domain closure. These results provide evidence for substantial conformational changes in the S1-M1 and S2-M4 linkers following agonist binding and channel opening, offering a conceptual frame to account for noncompetitive antagonism of AMPA receptors. PMID- 16242409 TI - NMDA receptor subunit composition controls synaptic plasticity by regulating binding to CaMKII. AB - Calcium entry through postsynaptic NMDA-Rs and subsequent activation of CaMKII trigger synaptic plasticity in many brain regions. Active CaMKII can bind to NMDA Rs, but the physiological role of this interaction is not well understood. Here, we test if association between active CaMKII and synaptic NMDA-Rs is required for synaptic plasticity. Switching synaptic NR2B-containing NMDA-Rs that bind CaMKII with high affinity with those containing NR2A, a subunit with low affinity for CaMKII, dramatically reduces LTP. Expression of NR2A with mutations that increase association to active CaMKII recovers LTP. Finally, driving into synapses NR2B with mutations that reduce association to active CaMKII prevents LTP. Spontaneous activity-driven potentiation shows similar results. We conclude that association between active CaMKII and NR2B is required for different forms of synaptic enhancement. The switch from NR2B to NR2A content in synaptic NMDA-Rs normally observed in many brain regions may contribute to reduced plasticity by controlling the binding of active CaMKII. PMID- 16242410 TI - Chromatin remodeling is a key mechanism underlying cocaine-induced plasticity in striatum. AB - Given that cocaine induces neuroadaptations through regulation of gene expression, we investigated whether chromatin remodeling at specific gene promoters may be a key mechanism. We show that cocaine induces specific histone modifications at different gene promoters in striatum, a major neural substrate for cocaine's behavioral effects. At the cFos promoter, H4 hyperacetylation is seen within 30 min of a single cocaine injection, whereas no histone modifications were seen with chronic cocaine, consistent with cocaine's ability to induce cFos acutely, but not chronically. In contrast, at the BDNF and Cdk5 promoters, genes that are induced by chronic, but not acute, cocaine, H3 hyperacetylation was observed with chronic cocaine only. DeltaFosB, a cocaine induced transcription factor, appears to mediate this regulation of the Cdk5 gene. Furthermore, modulating histone deacetylase activity alters locomotor and rewarding responses to cocaine. Thus, chromatin remodeling is an important regulatory mechanism underlying cocaine-induced neural and behavioral plasticity. PMID- 16242411 TI - Somatosensory integration controlled by dynamic thalamocortical feed-forward inhibition. AB - The temporal features of tactile stimuli are faithfully represented by the activity of neurons in the somatosensory cortex. However, the cellular mechanisms that enable cortical neurons to report accurate temporal information are not known. Here, we show that in the rodent barrel cortex, the temporal window for integration of thalamic inputs is under the control of thalamocortical feed forward inhibition and can vary from 1 to 10 ms. A single thalamic fiber can trigger feed-forward inhibition and contacts both excitatory and inhibitory cortical neurons. The dynamics of feed-forward inhibition exceed those of each individual synapse in the circuit and are captured by a simple disynaptic model of the thalamocortical projection. The variations in the integration window produce changes in the temporal precision of cortical responses to whisker stimulation. Hence, feed-forward inhibitory circuits, classically known to sharpen spatial contrast of tactile inputs, also increase the temporal resolution in the somatosensory cortex. PMID- 16242412 TI - Protein synthesis-independent plasticity mediates rapid and precise recovery of deprived eye responses. AB - Monocular deprivation (MD) for a few days during a critical period of development leads to loss of cortical responses to stimulation of the deprived eye. Despite the profound effects of MD on cortical function, optical imaging of intrinsic signals and single-unit recordings revealed that deprived eye responses and orientation selectivity recovered a few hours after restoration of normal binocular vision. Moreover, recovery of deprived eye responses was not dependent upon mRNA translation, but required cortical activity. Interestingly, this fast recovery and protein synthesis independence was restricted to the hemisphere contralateral to the previously deprived eye. Collectively, these results implicate a relatively simple mechanistic process in the reactivation of a latent set of connections following restoration of binocular vision and provide new insight into how recovery of cortical function can rapidly occur in response to changes in sensory experience. PMID- 16242413 TI - Progressive transformation of hippocampal neuronal representations in "morphed" environments. AB - Hippocampal neural codes for different, familiar environments are thought to reflect distinct attractor states, possibly implemented in the recurrent CA3 network. A defining property of an attractor network is its ability to undergo sharp and coherent transitions between pre-established (learned) representations when the inputs to the network are changed. To determine whether hippocampal neuronal ensembles exhibit such discontinuities, we recorded in CA3 and CA1 when a familiar square recording enclosure was morphed in quantifiable steps into a familiar circular enclosure while leaving other inputs constant. We observed a gradual noncoherent progression from the initial to the final network state. In CA3, the transformation was accompanied by significant hysteresis, resulting in more similar end states than when only square and circle were presented. These observations suggest that hippocampal cell assemblies are capable of incremental plastic deformation, with incongruous information being incorporated into pre existing representations. PMID- 16242415 TI - Integration of touch and sound in auditory cortex. AB - To form a coherent percept of the environment, our brain combines information from different senses. Such multisensory integration occurs in higher association cortices; but supposedly, it also occurs in early sensory areas. Confirming the latter hypothesis, we unequivocally demonstrate supra-additive integration of touch and sound stimulation at the second stage of the auditory cortex. Using high-resolution fMRI of the macaque monkey, we quantified the integration of auditory broad-band noise and tactile stimulation of hand and foot in anaesthetized animals. Integration was found posterior to and along the lateral side of the primary auditory cortex in the caudal auditory belt. Integration was stronger for temporally coincident stimuli and obeyed the principle of inverse effectiveness: greater enhancement for less effective stimuli. These findings demonstrates that multisensory integration occurs early and close to primary sensory areas and--because it occurs in anaesthetized animals--suggests that this integration is mediated by preattentive bottom-up mechanisms. PMID- 16242414 TI - Neural correlates of knowledge: stable representation of stimulus associations across variations in behavioral performance. AB - Behavioral responses to a sensory stimulus are often guided by associative memories. These associations remain intact even when other factors determine behavior. The substrates of associative memory should therefore be identifiable by neuronal responses that are independent of behavioral choices. We tested this hypothesis using a paired-associates task in which monkeys learned arbitrary associations between pairs of visual stimuli. We examined the activity of neurons in inferior temporal cortex as the animals prepared to choose a remembered stimulus from a visual display. The activity of some neurons (22%) depended on the monkey's behavioral choice; but for a novel class of neurons (54%), activity reflected the stimulus that the monkey was instructed to choose, regardless of the behavioral response. These neurons appear to represent memorized stimulus associations that are stable across variations in behavioral performance. In addition, many neurons (74%) were modulated by the spatial arrangement of the stimuli in the display. PMID- 16242416 TI - Staged correction of double discordance, situs inversus, and absence of true pulmonary arteries. AB - The case of a young boy with an extremely rare and complex cardiac malformation is described. Situs inversus, dextrocardia, double discordance with pulmonary atresia, absence of true pulmonary arteries, and systemico-pulmonary collateral arteries was the original diagnosis. Three preparatory operations were necessary to reconstruct and unifocalize the pulmonary vascular tree. At the age of 7 years complete correction with intraoperative stenting of the left neopulmonary artery was successfully performed. After more than 3 years from the definitive operation, the patient is alive with no physical limitations. PMID- 16242417 TI - Images in cardiothoracic surgery. Spontaneous rupture of the inferior thyroid artery leading to life-threatening mediastinal hematoma. PMID- 16242418 TI - Innovative monolateral approach for closed-chest atrial fibrillation surgery. AB - Surgical treatment of atrial fibrillation recently gained new popularity since the introduction of different energy sources as an alternative to the original cut-and-sew technique. Recently an innovative approach for closed-chest thoracoscopic epicardial pulmonary veins isolation has been described for patients suffering from lone atrial fibrillation. Nevertheless in an effort to further reduce the invasiveness of closed-chest atrial fibrillation surgery, we developed a novel monolateral approach for thoracoscopic arrhythmia surgery. PMID- 16242419 TI - Impact of resident duty hour standards on cardiothoracic residents and program directors. PMID- 16242420 TI - Should coronary artery bypass grafting be regionalized? PMID- 16242422 TI - Pulmonary arteriovenous malformations in children after the Kawashima operation. AB - BACKGROUND: Pulmonary arteriovenous malformations are a major cause of progressive late cyanosis in patients treated with cavopulmonary anastomoses. Previous experience suggests that exclusion of the hepatic venous effluent from the pulmonary circulation may cause the development of pulmonary arteriovenous malformations after the Kawashima operation in children with interrupted inferior vena cava with azygous continuation. METHODS: From January 1990 to November 2004, 21 children (median age, 2 years) with heterotaxy syndrome and interrupted inferior vena cava with azygous continuation underwent Kawashima operation. The average preoperative arterial oxygen saturation was 76% +/- 7% (range, 64% to 90%). RESULTS: Follow-up was complete in all survivors except 1 at a median duration of 4.5 years. Pulmonary arteriovenous malformations were diagnosed at a median of 5 years after Kawashima operation in 11 patients (58%). Completion Fontan operation has been performed in 15 (79%). Five children who underwent a completion Fontan procedure 1 to 1.5 years after Kawashima operation did not have pulmonary arteriovenous malformations. There were 2 late deaths due to chronic congestive heart failure 4 months and 7 years after Kawashima operation. Overall survival at 10 years is 90%. Univariate and multivariate analysis demonstrated presence of bilateral superior vena cavae (p = 0.002) and interval longer than 2 years between Kawashima operation and completion Fontan operation (p = 0.04) as predictors of developing pulmonary arteriovenous malformations. CONCLUSIONS: In most patients with heterotaxy and interrupted inferior vena cava with azygous continuation, clinical evidence of pulmonary arteriovenous malformations will develop after Kawashima operation if they are followed up long enough. Early redirection of the hepatic venous effluent to the pulmonary arterial circulation may prevent or lead to regression of pulmonary arteriovenous malformations, with low mortality and morbidity. PMID- 16242421 TI - Outcomes after the stage I reconstruction comparing the right ventricular to pulmonary artery conduit with the modified Blalock Taussig shunt. AB - BACKGROUND: Recent reports advocate that a right ventricular to pulmonary artery (RV-PA) conduit improves outcome after the stage I reconstruction. METHODS: We retrospectively compared the outcomes of all neonates who underwent a stage I reconstruction between January 1, 2002, and October 1, 2004, with use of the RV PA conduit and modified Blalock-Taussig shunt (mBTS) interspersed over this time period. RESULTS: In all, 149 infants underwent a stage I reconstruction (95 mBTS, 54 RV-PA) for hypoplastic left heart syndrome (HLHS) or variants. There was a preference for the RV-PA conduit in patients with aortic atresia (mBTS 30% versus RV-PA 67%, p < 0.01). There was no difference in surgical mortality (mBTS 14% versus RV-PA 17%, p = 0.67), time to extubation (mBTS 4.5 +/- 4.8 days versus RV PA 3.9 +/- 3.5 days, p = 0.47), or length of hospital stay (mBTS 25 +/- 29 days versus RV-PA 21 +/- 23 days, p = 0.52). There was an increased incidence of shunt reinterventions in the patients with the RV-PA conduit (mBTS 17% versus RV-PA 32%, p = 0.04). Patients with RV-PA conduit returned earlier for stage II reconstruction (mBTS 6.5 +/- 2.5 months versus RV-PA 5.6 +/- 1.7 months, p = 0.05). There was no difference in overall mortality (mBTS 32% versus RV-PA 30%, p = 0.45) with a median duration of follow-up of 18 +/- 8 months. CONCLUSIONS: Comparing shunt strategies (mBTS versus RV-PA) over the same time period, we found no difference in outcome. These data support the need for a larger prospective, randomized trial. PMID- 16242423 TI - Incorporation of the hepatic veins into the cavopulmonary circulation in patients with heterotaxy and pulmonary arteriovenous malformations after a Kawashima procedure. AB - BACKGROUND: In patients with polysplenia syndrome and azygous continuation of an interrupted inferior vena cava (IVC), pulmonary arteriovenous malformations (PAVMs) are relatively common after bidirectional cavopulmonary anastomosis (BCPA, Kawashima procedure). Resolution of PAVMs after hepatic vein (HV) inclusion into the cavopulmonary circulation has been reported, but there has been no systematic investigation of the effects of this therapy in a population of more than 3 patients. METHODS: We studied 16 patients with heterotaxy, univentricular congenital heart disease, and azygous continuation of the IVC who underwent incorporation of the HV into the cavopulmonary circuit for treatment of significant PAVMs after a Kawashima procedure. RESULTS: The median preoperative systemic arterial oxygen saturation (SsaO2) was 76% (65%-85%), compared with 89% (85% to 92%) early after BCPA. Among 15 early survivors, the median early postoperative SsaO2 was 76% (56%-85%). In 11 of the 15 survivors, SsaO2 rose to 90% or greater within a year and remained at 93% or greater at follow-up of 2.8 to 10 years. Four patients had persistent hypoxemia and residual PAVMs at follow up catheterization 1.5 to 8 years postoperatively; these patients had the most severe hypoxemia prior to HV inclusion, and in 2 the residual PAVMs were unilateral, with HV flow streaming to the contralateral lung, in which PAVMs had resolved. CONCLUSIONS: Hypoxemia resolved after cavopulmonary incorporation of the HV in the majority of our patients with PAVMs after the Kawashima operation, presumably due to a combination of PAVM resolution and elimination of hepatic venoatrial right-to-left shunting. These findings support the theory that development of PAVMs is facilitated by exclusion of HV effluent from the pulmonary circulation. PMID- 16242424 TI - Creation of a brachial arteriovenous fistula for treatment of pulmonary arteriovenous malformations after cavopulmonary anastomosis. AB - BACKGROUND: Pulmonary arteriovenous malformations (PAVMs) occur in approximately 20% of patients after unidirectional superior cavopulmonary anastomosis (CPA), and frequently after bidirectional CPA in patients with polysplenia syndrome. It is hypothesized that exclusion of a growth-modulating factor produced in the liver may predispose to PAVM formation. Resolution of PAVMs after inclusion of hepatic venous effluent into the cavopulmonary circulation has been reported. An upper extremity systemic arteriovenous (AV) fistula may be created to augment pulmonary blood flow and improve oxygenation in hypoxemic patients with CPA, but there has been no systematic investigation of the effects of such fistulas on PAVMs after CPA. METHODS: We studied 11 patients with PAVMs who underwent creation of a brachial AV fistula a median of 11 years after CPA. RESULTS: Eight patients had discontinuous pulmonary arteries or unilateral flow of a bidirectional CPA and were not considered good candidates for Fontan completion; the other 3 patients had polysplenia and unilateral hepatic venous streaming after Fontan completion. Three patients died of progressive complications of their heart disease 4 to 18 months after AV fistula creation. Pulmonary arteriovenous malformations resolved after creation of a brachial AV fistula in 4 of 5 surviving patients with unilateral flow of a superior CPA, but in none of 3 patients with polysplenia who had unilateral hepatic venous streaming after Fontan completion and PAVMs in the contralateral lung. CONCLUSIONS: These findings are consistent with the "hepatic factor" hypothesis, according to which the development of PAVMs is facilitated when an unidentified factor produced or metabolized in the liver does not reach the pulmonary circulation before traversing another capillary bed. Patients with unilateral superior CPA flow and PAVMs who are not considered candidates for Fontan completion may benefit from a brachial AV fistula. PMID- 16242425 TI - Superior durability of SynerGraft pulmonary allografts compared with standard cryopreserved allografts. AB - BACKGROUND: The ideal pulmonary valve replacement for children and adolescents remains elusive. Although favored by many surgeons, the cryopreserved pulmonary allograft tends to become rapidly incompetent and elicits an immune response. The SynerGraft process (Cryolife Inc, Kennesaw, GA) decellularizes a pulmonary allograft, leaving a scaffold of connective tissue. These grafts have been shown to decrease immune reactivity and become populated with host cells. Although theoretically these traits may improve durability, few data comparing SynerGraft processed allografts (SynAs) (Cryolife Inc) with standard cryopreserved allografts are available. METHODS: A single institution review was performed for all SynAs implanted from their introduction in 2001 to January 2003. Twenty-six patients with SynAs and 26 age and diagnosis-matched controls receiving cryopreserved allografts were evaluated. Subjects were analyzed for demographics, survival, reintervention, and echocardiographic findings. RESULTS: There were no significant differences between groups in age, weight, valve diameter, orthotopic and heterotopic allograft position, or follow-up. On echocardiogram there was no difference in initial degree of allograft insufficiency or gradient. However, at mean follow-up of 19 +/- 13 months, SynAs were significantly less regurgitant than cryopreserved allografts (p = 0.017). Although all gradients were low, a significant difference between SynAs (7.6 +/- 14 mm Hg) and cryopreserved allografts (14.6 +/- 15.6 mm Hg; p = 0.012) had emerged. Survival was identical at 85% (22 of 26). Rates of reintervention were similar at 7% (2 of 26) for cryopreserved allografts and 3.8% (1 of 26) for SynAs (p = 0.98). CONCLUSIONS: At intermediate follow-up, the SynA demonstrated greater durability with less insufficiency and lower gradients. These characteristics are important to many patients with complex congenital heart disease; however, long-term effects on survival and reintervention remain unknown. PMID- 16242426 TI - Early insertion of a pulmonary valve for chronic regurgitation helps restoration of ventricular dimensions. AB - BACKGROUND: This study assesses the results of early insertion of a pulmonary valve for chronic pulmonary regurgitation based on right ventricular volume measurements. METHODS: Valved conduits were prospectively inserted in 39 patients (aged 14 to 39 years) when the right ventricular (RV) end-diastolic volume index on magnetic resonance imaging (MRI) exceeded 150 mL/m2. Changes in morphology and function of the RV were prospectively analyzed by an MRI at 6 months postoperatively (available in 21 patients). RESULTS: There were no early or late deaths. All conduits showed good function at a median 15 month follow-up. Postoperative RV end-systolic and end-diastolic volumes showed a significant positive correlation (p = 0.005 and p < 0.0001), while postoperative left ventricular (LV) ejection fraction showed a significant negative correlation (p = 0.03) with preoperative RV end-diastolic volume index. Seven patients who achieved normal RV end-diastolic volume index (< or = 100 mL/m2) (group 1) when compared with 14 remaining patients (group 2) showed that they differed significantly with respect to their preoperative RV end-diastolic volume index (170.3 +/- 21.1 vs 203.6 +/- 35.6; p = 0.02) and postoperative LV ejection fraction (59.9 +/- 4.2 vs 54 +/- 7%, p = 0.03). CONCLUSIONS: Our results show that the improvement in ventricular dimensions and functions directly correlates with the timing of pulmonary valve insertion. Early insertion leads to normalization and late insertion leads only to improvement. These observations, along with a low morbidity for these reoperations, justify earlier reintervention in cases of chronic pulmonary regurgitation. A RV end-diastolic volume index of 150 mL/m2 seems to be a practical cutoff value to prescribe pulmonary valve insertion. PMID- 16242427 TI - Early experience with valve-sparing aortic root replacement in children. AB - BACKGROUND: Aortic valve-sparing root replacement has become an established treatment for adults with aneurysms of the ascending aorta, but there is limited experience in children, for whom the advantages of avoiding valve prostheses are particularly attractive. METHODS: A retrospective clinical study was undertaken to examine results of aortic valve-sparing operations in pediatric patients at a single institution. Clinical and echocardiographic data were obtained from chart review and outpatient follow-up examination. RESULTS: Between 1997 and 2004, 19 children underwent aortic valve-sparing root replacement. Mean age was 12.2 +/- 4.4 years, and mean weight was 51.5 +/- 22.9 kg. Median length of clinical follow up was 58 months (range, 5 to 91). Fifteen of 19 patients (78.9%) had Marfan syndrome and 1 had aortic root dilation late after the arterial switch operation. Mean preoperative root diameter was 4.7 +/- 0.6 cm, with an average Z score of 7.7 +/- 1.9. Fourteen patients (73.7%) underwent root remodeling, whereas 5 (26.3%) had a reimplantation procedure. One patient required concomitant mitral valve repair. There was no operative mortality and only 1 reoperation for bleeding (5.3%). Median length of hospital stay was 5 days (range, 3 to 12). At latest follow-up, 3 of 19 patients (15.8%) have required late aortic valve replacement. Fifteen patients have no or mild aortic valve insufficiency, and 1 has moderate but stable valve regurgitation. No patient with a reimplantation procedure has had a reoperation or more than mild insufficiency, and no patient has suffered endocarditis or thromboembolism. CONCLUSIONS: Valve-sparing aortic root replacement is a viable alternative to root replacement with mechanical or biological prostheses in children, and can be accomplished with minimal morbidity and mortality. Reimplantation procedures appear to have more durable results than root remodeling techniques, and should be strongly considered for pediatric patients with aortic root enlargement secondary to connective tissue disorders and other forms of congenital heart disease. PMID- 16242428 TI - Pediatric autograft aortic root replacement: a prospective follow-up study. AB - BACKGROUND: The autograft procedure offers children who require aortic valve replacement the advantage of an autologous valve that has growth potential and does not require anticoagulation. However, the autograft procedure is a double valve operation and its durability depends on the lifetime of both the autograft and the pulmonary valve substitute. We present our clinical experience with pediatric autograft aortic root replacement. METHODS: Between September 1988 and September 2003, 47 children (mean age, 8 years; standard deviation, 5 years; range, 3 months to 15 years) underwent autograft aortic root replacement. Perioperative characteristics and annual follow-up information were collected prospectively. RESULTS: The male to female ratio was 32 to 15. Eighty-nine percent of patients had congenital aortic valve disease, 47% of patients previously underwent cardiac surgery, and 43% had an aortic valve balloon dilatation. Concomitant left ventricular outflow tract enlargement was performed in 19 patients. In all cases the pulmonary valve was replaced using an allograft. There were no hospital deaths. Mean follow-up was 6.1 years (median 5.4; range, 1 month to 15 years; total of 284 patient years). During follow-up 3 patients died. Cumulative survival was 95% at 1 year and 93% at 12 years. One patient had endocarditis of the pulmonary allograft develop. Three patients required reoperation; two patients for allograft degeneration at 9.4 and 12.8 years, and 1 for combined autograft dilatation and allograft degeneration at 7.7 years postoperatively. Freedom from valve-related reoperation was 86% at 12 years. CONCLUSIONS: Pediatric autograft aortic root replacement is associated with acceptable mortality and reoperation rates in the first decade postoperatively. It allows most children to grow into adulthood without the need for anticoagulation and additional valve replacements. PMID- 16242429 TI - Coronary reimplantation after neoaortic reconstruction can yield better result in arterial switch operation: comparison with open trap door technique. AB - BACKGROUND: Accurate coronary reimplantation is the most important component in the arterial switch operation. It is especially demanding for the less experienced surgeons. We compared the result of the technique of coronary reimplantation after neoaortic reconstruction with that of the open trap door technique. METHODS: From March 1994 to June 2004, 103 consecutive patients underwent the arterial switch operation by one surgeon. Patients who underwent coronary artery transfer with other modified techniques were excluded. Diagnoses of 94 patients were transposition of the great arteries with intact ventricular septum (n = 50), transposition of the great arteries with ventricular septal defect (n = 26), and the Taussig-Bing anomaly (n = 18). An aortic arch anomaly was present in 13 patients. The median age of the patients was 12 days and the mean body weight was 3.5 kg. Coronary reimplantation after neoaortic reconstruction was applied to 34 patients (group I), and the open trap door technique was applied to the rest (group II). RESULTS: Preoperative data were similar in both groups. Four patients in group II required intraoperative revision of a transferred coronary artery, and 1 patient with an intramural left coronary artery in group I had a conversion to free grafting using the left subclavian artery. Overall early mortality was 17.0% (16 of 94). Mortality in group I (1 of 34; 2.9%) was significantly lower than in group II (15 of 60; 25.0%) (p = 0.008). The leading cause of death in group II was low cardiac output (n = 9). During the follow-up, an aortic regurgitation of greater than mild was detected in 2 patients in group II. CONCLUSIONS: Coronary reimplantation after neoaoartic reconstruction is an attractive method to minimize coronary artery transfer-related mortality or morbidity. PMID- 16242430 TI - Late results after PTCA for coronary stenosis after the arterial switch procedure for transposition of the great arteries. AB - BACKGROUND: The arterial switch operation has become the surgical approach of choice for d-transposition of the great arteries, but there is an increased awareness of adverse sequelae in some survivors. Long-term patency and normal function of the translocated coronary arteries must be achieved. It is reported that dependent of the prior coronary status, 3% to 11% of all survivors have proximal coronary stenosis or complete occlusion develop after arterial switch operations. However, treatment of these stenoses is still a matter of debate. Late results after percutaneous transluminal coronary angioplasty (PTCA) for coronary stenosis after the arterial switch operation for d-transposition of the great arteries are reported. METHODS: Seven children after arterial switch operation for d-transposition of the great arteries who had subsequently undergone PTCA for coronary stenosis were angiographically re-evaluated 3 to 15 months after the initial PTCA and again after 3 to 5 years. RESULTS: All children survived the initial PTCA procedure. There were no late deaths. The degree of stenosis before PTCA ranged from 74% to 97%; immediately after PTCA from 5% to 10%; at 3 to 15 months after PTCA from zero to 6%; and at 3 to 5 years after PTCA from zero to 3%. Three to 5 years after PTCA all children showed normal development of the treated coronary artery. CONCLUSIONS: Primary PTCA of stenotic proximal coronary arteries after the arterial switch procedure for d transposition of the great arteries seems to be an effective treatment with excellent long-term results. PMID- 16242431 TI - Invited commentary. PMID- 16242432 TI - Ascending aortic extension for enlargement of the aortopulmonary space in children with pulmonary artery stenosis. AB - BACKGROUND: Complex reconstruction of the aorta can be complicated by compression within the aortopulmonary space resulting in airway or pulmonary artery narrowing. Pulmonary artery compression is especially problematic in children with single ventricle physiology in which an increase in pulmonary vascular resistance may impair systemic venous flow and reduce cardiac output. METHODS: We operated on 7 patients (mean age, 2.9 years) with pulmonary artery stenosis presenting after a complex neonatal aortic reconstruction. All 7 patients underwent aortic extension with a polytetrafluoroethylene interposition graft and homograft patch angioplasty of the pulmonary artery to open the aortopulmonary space and relieve pulmonary artery narrowing. Five patients (hypoplastic left heart syndrome, n = 2; transposition of the great arteries with tricuspid atresia and aortic hypoplasia, n = 1; double outlet right ventricle with aortic hypoplasia, n = 2) had previously undergone first stage repairs for single ventricle morphology. Two of the patients had multiple interim procedures, including placement of bilateral pulmonary artery stents, prior to our repair. RESULTS: There was 1 early death secondary to fungal sepsis. Six patients were discharged from the hospital. There was 1 late, noncardiac death from aspiration pneumonia in a patient with a severe craniofacial defect. Follow-up echocardiograms in the intermediate term have demonstrated relief of pulmonary artery narrowing and unobstructed aortic flow. CONCLUSIONS: Aortic extension is an option in children with pulmonary artery compression of structures in the aortopulmonary space after complex aortic reconstruction. PMID- 16242433 TI - Carotid-subclavian artery index: new echocardiographic index to detect coarctation in neonates and infants. AB - BACKGROUND: In neonates and young infants (less than 3 months), coarctation may be missed or underestimated by echocardiography, especially with a patent ductus arteriosus or severe concurrent illness. A reliable noninvasive screening tool for coarctation would be useful for these patients. METHODS: From 1997 to 2003, echocardiographic evaluation was performed in 63 consecutive patients with coarctation (47 neonates and 16 infants) as well as in 23 controls (16 neonates and 7 infants). End-systolic measurements were obtained from 12 different sites of the aortic arch. RESULTS: In patients, the diameters of the ascending and descending aorta were comparable to controls, but the dimensions of the transverse arch were significantly smaller. The distances between the origins of the great vessels were longer in patients with coarctation than in controls. The ratio of the aortic arch diameter at the left subclavian artery, to the distance between the left carotid artery and the left subclavian artery, which we propose as the carotid-subclavian artery index, was significantly smaller in patients with coarctation. A cut-off point at 1.5 showed a sensitivity of 97.7% and 94.7%, and a specificity of 92.3% and 100%, for neonates and young infants, respectively. The positive predictive value to have coarctation was 97.7% and 100%, for neonates and infants, respectively. CONCLUSIONS: The carotid-subclavian artery index is a simply obtainable noninvasive screening parameter, showing high sensitivity and specificity for coarctation, and may be useful in unstable patients or in those with a patent ductus arteriosus in which coarctation may be overlooked. PMID- 16242434 TI - Invited commentary. PMID- 16242435 TI - Comparison of angioplasty and surgery for neonatal aortic coarctation. AB - BACKGROUND: The efficacy of balloon dilatation as primary treatment for neonatal aortic coarctation remains controversial. METHODS: A retrospective comparison between balloon angioplasty and surgery for the treatment of neonatal aortic coarctation was undertaken on 57 neonates younger than 40 days of age (angioplasty, 23 patients; surgery, 34 patients) treated between 1994 and 2004. RESULTS: Cohorts were similar with respect to the preinterventional variables of age, weight, upper extremity systolic blood pressure, coarctation gradient, degree of aortic arch hypoplasia, associated conditions, and mean follow-up (angioplasty, 36 months; surgery, 38 months). Among the angioplasty group, 13 patients (57%) required surgery, and 8 required a second balloon dilatation, of whom 3 patients had an aortic aneurysm. Among the surgery cohort, 6 patients experienced recurrence (18%) after either SFA (3) or XETE anastomosis repair (3). All were successfully treated with balloon angioplasty. Actuarial freedom from any intervention was significantly greater in the surgery cohort as was the degree of aortic arch growth. At latest follow-up, antihypertensive medication was required in 3 of 9 angioplasty patients (33%) and 2 of 27 surgery patients (7%). No repeat intervention was required in the 13 patients who underwent angioplasty followed by surgery. CONCLUSIONS: Primary angioplasty is palliative treatment for neonatal aortic coarctation, but it is the treatment of choice for recurrence after surgery. Surgery for neonatal aortic coarctation is associated with fewer reinterventions, improved aortic arch growth, no aortic aneurysm formation, and decreased need for antihypertensive medication when compared with neonates treated primarily with balloon angioplasty. PMID- 16242436 TI - Cor triatriatum: presentation, diagnosis and long-term surgical results. AB - BACKGROUND: Cor triatriatum sinister is a rare, surgically correctable, congenital cardiac anomaly. Fewer than 250 cases have been reported. It can occur as an isolated defect (classic) or in association with other congenital cardiac anomalies (atypical). METHODS: A retrospective review of 28 consecutive patients diagnosed at the Royal Children's Hospital in Melbourne, Australia during a 22 year span from 1981 to 2003. RESULTS: There were 13 men (46%) and 15 women (54%). Fifteen patients were less than 1 year of age (neonates, [n = 7], 25%; infants, [n = 8], 29%). 17 patients (61%) had a communication between the right atrium and either the proximal or distal chamber. Fifteen patients (54%) had atypical cor triatriatum. Median age at presentation was 6 months (range, 0.6 to 240). Twenty four patients (86%) had presented by 5 years of age. Five patients (18%) underwent emergency surgery. Median age at operation was 7 months (range, 1 to 243). Twenty-seven patients (96%) underwent preoperative transthoracic echocardiography. Nine patients (32%), all with atypical cor triatriatum, required cardiac catheterization for diagnosis. Twenty-seven patients (96%) were correctly diagnosed before treatment. Twenty-seven patients (96%) were treated surgically. The defect was approached through the right atrium in 26 patients (93%). There was 1 early death and 1 patient died 10 years after repair. Follow up was 86% complete. At a median follow-up of 98 months (range, 0.2 to 284), all patients including those with atypical cor triatriatum were in New York Heart Association's function class 1. Post-repair survival was 96% and 88% at 5 and 15 years, respectively. CONCLUSIONS: Surgery offers good early and long-term results for both classic and atypical cor triatriatum. PMID- 16242437 TI - Steroid supplementation: a legitimate pharmacotherapy after neonatal open heart surgery. AB - BACKGROUND: An inflammatory response together with multiple organ failure subsequent to cardiopulmonary bypass is especially prominent in neonates. The behavior of glucocorticoids during this period in these patients is not known. If adrenal insufficiency should exist, it could considerably compromise postoperative recovery. METHODS: Twenty neonates undergoing biventricular repair were enrolled. Ten patients were assigned to receive hydrocortisone treatment and the other 10 to receive placebo. The treatment group received stress-dose hydrocortisone sodium succinate after discontinuation of cardiopulmonary bypass: 0.18 mg.kg(-1).hr(-1) for 3 days, 0.09 mg.kg(-1).hr(-1) for 2 days, and 0.045 mg.kg(-1).hr(-1) for 2 days. The placebo was 5% glucose solution. RESULTS: Patients had adrenal insufficiency (cortisol < 5 microg/dL) from 24 to 72 hours in the placebo group. This was associated with a simultaneous reduction of left ventricular shortening fraction (p < 0.0001, analysis of variance; p = 0.0203, Student's t test), the necessity to increase inotropic agents (p = 0.043, analysis of variance), and an increase in serum lactate level (p = 0.049, Student's t test). During this period, serum cortisol level was maintained above the normal level (> 23 microg/dL) in the hydrocortisone group. The placebo group had a greater positive fluid balance (p = 0.027, Student's t test) and greater total body edema in the immediate postoperative period (p = 0.065, Student's t test). Blood oxygenation constantly improved, and the duration on mechanical ventilation was shorter (83.5 +/- 42.1 versus 138.2 +/- 89.7 hours; p = 0.098) in the hydrocortisone group. CONCLUSIONS: Adrenal insufficiency may occur after neonatal open heart surgery. Stress-dose hydrocortisone supplementation blunts other organ dysfunction and can be considered a legitimate pharmacotherapy in this cohort. PMID- 16242438 TI - Hypothermic circulatory arrest enables aortic valve replacement in patients with unclampable aorta. AB - BACKGROUND: Atheroembolic complications associated with clamping a severely diseased ascending aorta during aortic valve replacement may result in unacceptable mortality and morbidity. Different management options include hypothermic circulatory arrest to replace the aortic valve, an aortic endarterectomy, or tube graft replacement of the aorta to allow safe application of cross-clamp before aortic valve replacement. METHODS: From 1998 to 2004, 70 patients who underwent aortic valve replacement had an aorta that was unclampable. Median age was 76 years; 33 (47%) were women; 46 (66%) had concomitant coronary artery bypass grafting; 9 (13%) had concomitant mitral valve surgery; and 4 (6%) were reoperations. Hypothermic circulatory arrest was used to replace the aortic valve alone, to do an aortic endarterectomy, or replace the ascending aorta with a tube graft. RESULTS: Operative mortality was 4%. There were 8 (11%) strokes and 1 (1.4%) transient ischemic attack. Statistical analysis showed no association between circulatory arrest period and occurrence of adverse cerebral events. There was no significant difference among the three groups when operative mortality and cerebral events were compared. CONCLUSIONS: Hypothermic circulatory arrest is an important adjunct that allows aortic valve replacement to be performed with an acceptable mortality but with an increased risk of cerebral event in this high-risk and elderly group of patients. PMID- 16242439 TI - No-clamp technique for valve repair or replacement in patients with a porcelain aorta. AB - BACKGROUND: Patients requiring valvular heart surgery may have circumferential calcification of the ascending aorta. A variety of creative procedures have been described for managing this "porcelain aorta." We describe a technique based on replacement of the ascending aorta and proximal arch under profound hypothermic circulatory arrest, followed by the valve procedure. METHODS: Twenty-five consecutive patients with a porcelain aorta were referred for heart valve surgery. In every case the aorta was replaced under circulatory arrest before the valve procedure. Postoperative morbidity, mortality, and univariate risk factors for death were calculated. Fisher's exact test defined significant perioperative variables with a p value less than 0.05. RESULTS: Of 25 patients, 23 (92%) survived the surgery to hospital discharge. One patient had a stroke (4%) and 2 patients (8%) required reexploration for bleeding. Risk factors for perioperative death by univariate analysis included age more than 78 years (p < 0.009), cardiopulmonary bypass time longer than 200 minutes (p < 0.0001), reexploration for bleeding (p < 0.02), need for intra-aortic balloon pump support (p < 0.001), and postoperative gastrointestinal complications (p < 0.001). CONCLUSIONS: Valve replacement or repair in the patient with a porcelain aorta can be safely accomplished with a technique based on aortic replacement under circulatory arrest. Elderly patients requiring extensive procedures and prolonged periods on bypass have a substantially increased risk for postoperative complications and death. PMID- 16242440 TI - Quality of life after heart valve surgery with prolonged intensive care. AB - BACKGROUND: A small proportion of patients undergoing heart valve operations require prolonged intensive care after surgery. Little is known about the quality of life that such patients attain after hospital discharge. METHODS: All consecutive patients who underwent primary heart valve surgery from 1998 to 2003 and required 8 days or more of treatment in an intensive care unit (ICU) were included (n = 225). At follow-up on August 31, 2004, 154 of these patients were alive. A cohort (n = 154) matched for sex, age, type of procedure, and week of operation, with an uncomplicated postoperative course (ICU stay of 2 days or less), served as the control group. All patients received the Medical Outcomes Study Short-Form 36, the Nottingham Health Profile, and the Hospital Depression and Anxiety scale to evaluate their quality of life. RESULTS: Survival at 5 years in the total ICU group was 68% (154 of 225). According to SF-36, the ICU study cohort reported poorer physical health but equal mental health compared with controls. On the Nottingham Health Profile, the ICU group reported more problems in all domains except emotional reactions and sleep. There was no difference in anxiety or depression between the groups. The ICU patients were in more advanced New York Heart Association functional classes preoperatively and postoperatively. No patient in the ICU study cohort regretted undergoing the operation, and 80% experienced improvement after surgery. CONCLUSIONS: This study showed reduced quality of life in terms of physical health and equal mental health in patients who required prolonged intensive care after heart valve surgery compared with controls without complications. PMID- 16242441 TI - Mitroflow synergy prostheses for aortic valve replacement: 19 years experience with 1,516 patients. AB - BACKGROUND: Pericardial prostheses have been used as valvular substitutes since 1975. They combine excellent hemodynamic characteristics with a low risk of valve failure during long-term observation. The aim of this article is to describe the clinical long-term performance of the Mitroflow Synergy pericardial valve (Sorin Group Inc, Mitroflow Division, Vancouver, Canada) in the aortic position for as long as 19 years. METHODS: Data were obtained between February 1985 and April 2004 from patients with aortic heart valve replacements (n = 1,464) or from patients with replacements of existing prosthetic aortic valves (n = 52). The age group distributions are less than 70 years (n = 175); 70 to 74 years (n = 462); 75 to 79 years (n = 532); 80 to 84 years (n = 273); and greater than or equal to 85 years (n = 74). The cause of the aortic valve lesions was combined (insufficiency and stenosis) in the majority of patients (62.4%). Concomitant procedures were performed in 897 patients (59.2%) and coronary artery bypass grafting was the most common (53.5%). Mean follow-up was 5.5 +/- 0.09 years. Total follow-up was 8,408 patient-years. RESULTS: The early mortality (30 days) was 6.6% (n = 99) and late deaths were 60.8% (n = 921). Actuarial event-free rates at 5, 10, and 15 years of follow-up are given as mean +/- standard error for endocarditis: 96.9 +/- 0.5, 92.8 +/- 1.2, and 92.0 +/- 1.4, respectively; embolism: 96.7 +/- 0.6, 88.8 +/- 1.7, and 82.9 +/- 3.5, respectively; bleeding: 98.5 +/- 0.4, 97.2 +/- 0.6, and 94.4 +/- 1.7, respectively; structural valve deterioration: 99.0 +/- 0.3, 82.8 +/- 2.2, and 62.8 +/- 5.8, respectively; and reoperation: 98.2 +/- 0.4, 79.2 +/- 2.4, and 63.4 +/- 5.3, respectively. The rate of endocarditis, structural valve degeneration, and reoperation was lower in patients 75 years of age and older compared with younger patients, whereas embolism occurred more frequently in elderly than in younger patients. CONCLUSIONS: Adverse events after implantation of Mitroflow aortic bioprosthesis rarely occurred during the first 5 years after valve replacement. The results of the prosthesis indicates reliable long-term morbidity rates and good durability in patients 75 years of age and older. PMID- 16242442 TI - Invited commentary. PMID- 16242443 TI - Intraoperative effects of the coapsys annuloplasty system in a randomized evaluation (RESTOR-MV) of functional ischemic mitral regurgitation. AB - BACKGROUND: Functional ischemic mitral regurgitation (MR) frequently arises after myocardial infarction; it is characterized by annular enlargement or lateral displacement of the subvalvular apparatus. Coapsys is a ventricular-annular remodeling device designed to treat functional ischemic MR; it does not require cardiopulmonary bypass. Initial intraoperative results of the RESTOR-MV randomized clinical trial are presented. METHODS: Patients referred for coronary artery bypass grafting with preoperative MR grade of 2 or greater were studied, excluding those with structural valve abnormalities. The Coapsys device, which consists of two epicardial pads connected by a flexible cord, was surgically implanted in 19 patients. Under epicardial echocardiographic guidance, the cord was passed through the left ventricle and tightened externally to improve leaflet coaptation and stabilize the ventricular wall; tightening was conducted with color flow Doppler imaging. RESULTS: Patients were 64.5 +/- 9.2 years old with an ejection fraction of 0.383 +/- 0.089 and received 2.7 +/- 1.1 grafts. Intraoperative MR grade was 2.7 +/- 0.8 after induction and was reduced to 0.4 +/ 0.7 after implantation (p < 0.0001). Mean epicardial dimension was reduced from 8.5 +/- 1.2 to 6.4 +/- 0.9 cm (p < 0.0001). Intraoperative MR was reduced in 95% (18 of 19) of patients, and 84% (16 of 19) had MR grade 1 or less after implantation. All implants were performed without cardiopulmonary bypass or conversion to standard annuloplasty. No hemodynamic compromise or structural damage to the mitral apparatus was noted. Significant acute remodeling was noted in the left ventricular dimensions. CONCLUSIONS: In patients without structural valve disease, the Coapsys device acutely reduces functional MR. Further randomized evaluation will assess long-term stability and compare it with standard annuloplasty techniques. PMID- 16242444 TI - Surgical treatment of cardiac papillary fibroelastoma: a single center experience with eighty-eight patients. AB - BACKGROUND: Cardiac papillary fibroelastoma is a rare benign tumor that can cause thromboembolism. We have found no large surgical series describing its treatment and outcome. METHODS: A retrospective review of all patients treated surgically for this tumor from 1985 to 2002. RESULTS: There were 88 patients with a mean age of 62 +/- 16 years. Sixty-two (71%) were male. Cardiac papillary fibroelastoma was a primary indication for surgery in 47 (group 1, 53%) and an incidental finding in 41 (group 2, 47%). The common clinical symptoms were neurologic (group 1) and cardiac (group 2). Cardiac valves were predominantly involved (77%); the aortic valve was the most affected (52%). Other common sites were the left ventricular outflow tract (18%) and anterior mitral leaflet (11%). All heart valves were involved in one patient. Seventy-three patients (83%) had shave excision and 8 (9%) excision with valve repair. Of 5 (6%) valve replacements, 2 were for concurrent degenerative valve disease. Concomitant procedures included repair or replacement of another valve (32%), CABG (28%), and septal myectomy (19%). Surgical mortality occurred in 1 patient (2.1%) in group 1 who had concomitant lung resection for bronchiolitis obliterans. There was no tumor recurrence, and no tumor-related late morbidity or mortality at a mean follow-up of 3 years. CONCLUSIONS: Cardiac papillary fibroelastoma has a propensity to affect the anatomically contiguous structures of the aortic valve, left ventricular outflow tract, and anterior mitral leaflet. Surgical treatment by simple shave excision is low risk and can achieve good results. PMID- 16242445 TI - Is reoperation still a risk factor in coronary artery bypass surgery? AB - BACKGROUND: Hospital mortality for reoperative coronary artery bypass grafting (CABG) is approaching that of primary CABG. This raises two questions: (1) has experience neutralized the risk of reoperation attributable to its greater difficulty, or (2) has experience neutralized the risk attributable to the higher risk profile of reoperative patients?. METHODS: From 1990 to 2003, 21,568 CABG procedures were performed, of which 4,518 (21%) were reoperations: 3,919 first, 552 second, 43 third, 3 fourth, and 1 fifth. Reoperative patients had a higher risk profile than primary patients, with more vascular disease, left ventricular dysfunction, and coronary artery disease (all p < 0.0001). Logistic regression was used to identify factors associated with hospital death and to develop a propensity score for reoperation, which was used to (1) adjust multivariable analyses of death and (2) compare outcomes in matched patients. RESULTS: Hospital mortality was 4.3% (168 of 3,919) for first reoperation, 5.1% (28 of 552) for second, and 6.4% (3 of 47) for third or more, compared with 1.5% (263 of 17,050) for primary operations. Risk of both primary and reoperative CABG decreased with experience (p > 0.0002); however, reoperative risk fell markedly in the mid 1990s. In both the overall and matched-pairs analyses, reoperation was a risk factor before 1997 (p < or = 0.008), but not after (p = 0.2). Reoperation within 1 year of previous CABG increased risk (p < 0.0001). Risk attributable to left ventricular dysfunction decreased with experience (p = 0.05). CONCLUSIONS: Hospital mortality for reoperative CABG has been consistently higher than for primary operation, but this difference has narrowed considerably. Patient characteristics, not reoperation itself, now have greater influence. PMID- 16242446 TI - Evidence of nitric oxide produced by the internal mammary artery graft in venous drainage of the recipient coronary artery. AB - BACKGROUND: The endothelium of the internal mammary artery produces nitric oxide in greater quantity than other vessels employed in revascularization of the ischemic myocardium. The aim of this study was to measure the concentration of stable metabolite (nitrite) of the endothelium-derived nitric oxide in the venous drainage (anterior interventricular vein) of the recipient coronary artery, which was the left anterior descending branch. The sampling was carried out before and after anastomosis completion. METHODS: Nitrite levels in the anterior interventricular vein, before and after anastomosis completion, in the left internal mammary artery free flow, and in the subclavian vein were measured. Fluroscopy after 4-hydroxycoumarin nitrozation was utilized to measure nitrite content of blood samples in 50 consecutive, partly heparinized patients undergoing off-pump coronary bypass surgery. Nitrate content of all samples was removed by Cadmium pearls. RESULTS: One hundred and sixty-four samples taken from 41 patients were feasable to analyze. A significant increase of nitric oxide (nitrite) level was found in the anterior interventricular vein, when comparing concentrations measured before and after the anastomosis between the left internal mammary artery and the left anterior descending artery. Mean values in the anterior inteventricular vein before and after anastomosis completion were as follows: 44.8 microMol (SD 4.9) and 70.7 microMol (SD 8.1), respectively. CONCLUSIONS: The increased production of nitric oxide by the internal mammary arterial graft may provide a perpetual vasodilatory response and partially protect the distal coronary vessel from atherosclerosis. PMID- 16242447 TI - Changing incidence, type, and natural history of conduction defects after coronary artery bypass grafting. AB - BACKGROUND: Cardiac conduction defects occur after cardiac surgery. We hypothesized that population aging and increased use of beta-blockers would increase the incidence of new conduction defects after coronary surgery. METHODS: We examined the medical records of 800 coronary artery bypass grafting (CABG) patients (400 from 1991 and 400 from 2001). Exclusion criteria included the following: preexisting conduction defect, permanent pacemaker, and perioperative atrial fibrillation, leaving 303 and 269 patients, respectively, included in the two study years. The incidence, type, and persistence of new conduction defects were determined from the preoperative, postoperative, and the predischarge electrocardiogram. Multivariate analysis identified predictors of new defects. RESULTS: Study populations were well-matched. There was a marked decrease in the incidence of new postoperative conduction defects from 1991 (19%) to 2001 (6%). There was also a change in the most frequently occurring block, from a right bundle-branch-block in 1991 (10%) to first-degree atrioventricular block (3%) in 2001. Finally, conduction defects in 1991 were more transient. While 19% of 1991 patients showed a conduction defect early postoperatively, only 9% were persistent. In 2001, the incidence of conduction defects at discharge (7%), was equivalent to that early postoperatively (6%). Predictors of new conduction defects included year of operation, age, intraaortic balloon counterpulsation, number of vessels bypassed, and crystalloid cardioplegia. CONCLUSIONS: Our results were the opposite of those predicted. Our report identifies a changing incidence, type, and natural history of conduction defects after CABG. Our comparison demonstrated a decrease in the incidence of new conduction defects, as well as a qualitative change in the defects identified. Multivariate analysis provided predictors of new conduction defects after CABG. PMID- 16242448 TI - Results of coronary artery endarterectomy and coronary artery bypass grafting for diffuse coronary artery disease. AB - BACKGROUND: Coronary artery endarterectomy with coronary artery bypass grafting for diffuse coronary artery disease has been associated with increased morbidity and mortality. We evaluated our institutional experience to redefine the role of coronary endarterectomy for diffuse coronary artery disease. METHODS: From 1985 to 2002 isolated coronary artery endarterectomy with coronary artery bypass grafting was performed in 1,478 consecutive patients. The short-term outcomes were compared with concurrent series of conventional coronary artery bypass graft surgery, and risk factors for adverse outcomes after coronary endarterectomy were identified. RESULTS: Patients in the coronary endarterectomy group were of higher risk with increased incidence of comorbidities and three-vessel coronary disease. The operative mortality (3.2% versus control 2.2%; p = 0.03) and the incidence of major postoperative morbidity (not significant) were comparable between the groups. Prolonged cardiopulmonary bypass time, recent acute myocardial infarction, redo surgery, and poor ventricular function were important predictors of in-hospital mortality. Vessel endarterectomized, technique of endarterectomy, and cardiopulmonary bypass versus off-pump technique did not alter results. At long-term follow-up, 5-year and 10-year survivals were 83% +/- 5%, and 74% +/- 3%, respectively, and freedom from angina at 5 and 10 years was 75% +/- 5%, and 69% +/- 4%, respectively, with 96% of survivors in New York Heart Association class II. CONCLUSIONS: In selected patients with diffuse coronary artery disease, coronary endarterectomy can be used as a tool for myocardial revascularization. The operative mortality and major morbidity were comparable or similar to coronary artery bypass grafting, and short-term and long-term results were favorable. PMID- 16242449 TI - Invited commentary. PMID- 16242450 TI - Stroke after coronary artery bypass grafting. AB - BACKGROUND: Despite the continuing improvements in surgical and cardiopulmonary bypass techniques during cardiac surgery, stroke remains a devastating complication. This study aimed to identify the preoperative and intraoperative risk factors for developing a perioperative stroke in patients undergoing coronary artery bypass graft surgery on cardiopulmonary bypass. METHODS: A total of 4,380 consecutive patients who received isolated coronary artery grafting on cardiopulmonary bypass between 1992 and 2002 were included. The sample contained three cardiopulmonary bypass temperature strategies: hypothermic (< 31 degrees C, n = 1,853), tepid (32-35 degrees C, n = 1,088), and normothermic (> 36 degrees C, n = 1,439). Outcome measures reported include stroke incidence, 30-day mortality, and hospital length of stay. RESULTS: The incidence of stroke was 1.2% (n = 51). Stroke patients were older, were more likely to be diabetic, hypertensive, have creatinine levels greater than 0.12 mmol/L, and have a history of stroke than those who did not have stroke (p < 0.05). Multivariate logistic regressions identified diabetes (p = 0.01), history of stroke (p = 0.04), and older age (p = 0.05) as independent predictors of stroke for all patients. The 30-day mortality for stroke patients was ten times greater than that of those who did not suffer stroke (17.6 vs 1.7%). CONCLUSIONS: Diabetes, history of stroke, and older age were identified as risk factors for stroke after coronary bypass; the temperature at which cardiopulmonary bypass was performed was not significant. PMID- 16242451 TI - Adverse 30-day outcomes after cardiac surgery: predictive role of intraoperative myocardial acidosis. AB - BACKGROUND: Regional myocardial acidosis in patients undergoing cardiac surgery has been shown to be reflective of regional myocardial ischemia. This study elucidates the relationship between intraoperative regional myocardial acidosis and 30-day postoperative outcomes after cardiac surgery. METHODS: Intramyocardial tissue pH in the anterior and posterior left ventricular walls was measured in 397 adult patients undergoing valve replacement or coronary revascularization surgery between 1987 and 2001. Dedicated nurses and research assistants prospectively collected preoperative, intraoperative, and outcomes data. Regional myocardial acidosis was defined in terms of pH thresholds identified by recursive partitioning. Adverse 30-day outcome, defined as death or any one of six complications, was the dependent variable in a multivariate logistic regression analysis. A morbidity score was developed on the basis of the sensitivity of each of the six complications in predicting death, and was the dependent variable in a multivariate linear regression analysis. RESULTS: During the period of aortic clamping, a mean intramyocardial tissue pH less than 6.85 was identified to be significant by recursive partitioning, and was encountered in either the anterior or posterior left ventricular wall in 85.4% of patients. After adjusting for preoperative and intraoperative variables, this pH threshold was found to be significantly associated with increased adverse outcomes within 30 days after surgery (p = 0.045). It was also significantly associated with increase in the morbidity score (p = 0.05). CONCLUSIONS: Regional myocardial acidosis of a magnitude frequently encountered during aortic clamping is an independent determinant of adverse 30-day outcomes after cardiac surgery. Its reversal by pH guided myocardial management has the potential of improving postoperative patient outcomes. PMID- 16242452 TI - Cardiac troponin I release after coronary artery bypass grafting operation: effects on operative and midterm survival. AB - BACKGROUND: Markers of myocardial necrosis are usually elevated in patients who have undergone a coronary bypass operation with cardiac arrest. The preferred marker in detecting acute myocardial ischemia is cardiac troponin I (cTnI). However, its ability to predict short-term and, particularly, midterm outcome after coronary bypass operations is uncertain. METHODS: Two hundred thirty unselected patients undergoing surgical revascularization had cTnI measured preoperatively and 11 times postoperatively. Receiver operating characteristic curves were constructed using cTnI postoperative peak values in order to assess the prognostic sensitivity and specificity of the test. The cut-off value of 13 ng/mL was used to assess the prognostic significance of the peak cTnI postoperative release for short-term and midterm outcomes. RESULTS: One hundred forty-six patients (63.5%) had postoperative cTnI peak values less than 13 ng/mL (mean peak value, 6.6 +/- 3.1 ng/mL) and 84 patients (36.5%) had postoperative cTnI peak values greater than 13 ng/mL (mean peak value, 45.5 +/- 59.9 ng/mL). Patients with peak cTnI greater than 13 ng/mL were older and had higher preoperative cTnI values. They required both longer cross-clamp time and CPB time. Moreover, hospital death in the cTnI greater than 13 ng/mL group (9.5% versus 0.7%, p = 0.0009) was significantly higher. Multivariate analysis showed that cTnI greater than 13 ng/mL was the only independent predictor of hospital death (odds ratio 10.33, p = 0.04) and hospital death from cardiac causes. A 2 year follow-up demonstrates that cTnI postoperative release had no influence on midterm mortality and hospitalization for due to cardiac illness. CONCLUSIONS: Cardiac troponin I is a valuable marker for immediate myocardial damage after coronary bypass operations. Its postoperative release does not predict midterm outcome. PMID- 16242453 TI - Incidence of sternal infection in diabetic patients undergoing bilateral internal thoracic artery grafting. AB - BACKGROUND: Bilateral internal thoracic artery (BITA) bypass grafts have advantages over single internal thoracic artery (SITA) bypass grafts in the medium term, particularly in diabetics. However, the perceived higher sternal complication rates seen in diabetics have led many surgeons to avoid the use of BITA surgery. The aim of our study was to assess the validity of this approach by assessing the incidence of sternal infections over a 10-year period in one institution. METHODS: A retrospective analysis was made of our coronary artery bypass graft (CABG) patients over a 10-year period (7,581 patients). Nine hundred and twenty-two of the patients were diabetics (261 insulin-dependent diabetes mellitus [IDDM]). Of the insulin-dependent diabetics, 166 had SITA, and 95 had BITA grafts. RESULTS: There was no significant difference in this subgroup in terms of gender, preoperative angina, dyspnea class, left ventricular function, and number of distal anastomoses. Comparing the rates of sternal wound complications of SITA and BITA in IDDM are the following: (1) superficial sternal infection, 6.6% in SITA, 1.1% in BITA (p = 0.04); (2) deep sternal infection, 1.2% in SITA, 3.2% in BITA (p = 0.27); (3) sternal dehiscence, 1.2% in SITA, 3.2% in BITA (p = 0.27). CONCLUSIONS: Our data do not support the perception that BITA grafting increases the risk of sternal complications in insulin-dependent diabetic patients. PMID- 16242454 TI - Effects of resveratrol in storage solution on adhesion molecule expression and nitric oxide synthesis in vein grafts. AB - BACKGROUND: Endothelial injury in human saphenous vein grafts may occur during harvesting and storage, which may have an adverse effect on coronary artery bypass grafting outcome. In this study, we sought to determine whether resveratrol, a natural antioxidant enriched in grape, can limit endothelial activation and reduce endothelial injury in human saphenous vein grafts. METHODS: Human saphenous vein grafts, obtained from 8 patients and divided into two equal groups of control and study specimens, were stored in either heparinized blood (group A) or heparinized blood containing 50 microg/mL resveratrol (group B) for 1 hour at room temperature. Specimens were analyzed by Western blotting to quantify intercellular adhesion molecule-1, vascular cell adhesion molecule-1, and inducible nitric oxide synthase-2 expression, as well as tissue cyclic guanylate monophosphate levels. Myeloperoxidase activity, a marker of neutrophil sequestration in human saphenous vein grafts, was also measured in each group. RESULTS: Intercellular adhesion molecule-1 expression (1,674 +/- 332 versus 559 +/- 282; p = 0.003), vascular cell adhesion molecule-1 expression (753 +/- 183 versus 472 +/- 151; p = 0.025), and myeloperoxidase activity (7.00 +/- 1.05 versus 1.33 +/- 0.45 nm/min; p = 0.004) were significantly lower in group B. In contrast, inducible nitric oxide synthase-2 expression (548 +/- 237 versus 2,234 +/- 726; p = 0.004) and tissue cyclic guanylate monophosphate levels (2.02 +/- 0.53 versus 5.61 +/- 0.89 pmol/mL; p = 0.001) were significantly higher in group B. CONCLUSIONS: Resveratrol reduced upregulation of leukocyte-endothelial cell adhesion molecule expression in human saphenous vein graft endothelium and decreased neutrophil adhesion to saphenous vein graft endothelium. Resveratrol also augmented inducible nitric oxide synthase-2 expression and increased cyclic guanylate monophosphate levels. These results suggest that resveratrol might improve vascular homeostasis and reduce endothelial injury during the hypoxic storage period of human saphenous vein grafts for coronary artery bypass grafting. PMID- 16242455 TI - Increasing transplanted cell survival with cell-based angiogenic gene therapy. AB - BACKGROUND: The majority of cells transplanted into infarcted myocardium do not survive. Maximizing cell survival should maximize the efficacy of cell transplantation for myocardial repair. We evaluated the role of apoptosis in cell loss after transplantation and the effect of angiogenesis on apoptosis and overall cell survival. METHODS: Female Lewis rats underwent myocardial cryoinjury 3 weeks before transplantation with male heart cells (a mixed culture of cardiomyocytes, smooth muscle cells, endothelial cells, and fibroblasts), vascular endothelial growth factor-transfected heart cells, skeletal myoblasts, vascular endothelial growth factor-transfected skeletal myoblasts (n = 6 each), or medium (control, n = 5). One week later, transplanted cell survival and apoptosis were quantitated by real-time polymerase chain reaction for Y chromosomal deoxyribonucleic acid, terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling assay and deoxyribonucleic acid fragmentation. RESULTS: Approximately one third of heart cells and skeletal myoblasts survived 1 week after transplantation, and one half of vascular endothelial growth factor-transfected heart cells and skeletal myoblasts survived to this time (p < 0.05). Apoptosis was greatest in heart cell and skeletal myoblast-transplanted hearts (p < 0.05), reduced in the vascular endothelial growth factor-transfected groups (p < 0.05) and lowest in controls. CONCLUSIONS: Ischemia and apoptosis both contribute to cell loss after transplantation. Transfection with vascular endothelial growth factor induced angiogenesis, which reduced both ischemic and apoptotic cell death. Our findings suggest that further strategies to reduce apoptosis may enhance the efficacy of cell transplantation in myocardial repair. PMID- 16242456 TI - Effects of mycophenolate mofetil on cardiac allograft survival and cardiac allograft vasculopathy in miniature swine. AB - BACKGROUND: Chronic rejection, as manifested by cardiac allograft vasculopathy, remains the leading cause of late graft failure in heart transplant recipients. Despite recent clinical trials, the efficacy of mycophenolate mofetil in preventing human cardiac allograft vasculopathy remains controversial. We investigated whether mycophenolate mofetil would prevent cardiac allograft vasculopathy and prolong cardiac allograft survival in our well-established miniature swine model of heart transplantation. METHODS: Hearts disparate at the major histocompatibility complex class I locus were heterotopically transplanted into miniature swine recipients treated with a 12-day course of mycophenolate mofetil (n = 3) or cyclosporine A (n = 3). Allograft survival, acute rejection, and chronic rejection were monitored in the two groups. RESULTS: Hearts transplanted with 12 days of cyclosporine were rejected between 46 and 61 days, whereas two of the three hearts transplanted with mycophenolate mofetil remained beating beyond 120 days (p = 0.02). At necropsy, there was a 4.9% mean prevalence of cardiac allograft vasculopathy in the mycophenolate mofetil group as compared with 16.6% in the cyclosporine group (p = 0.03). Cardiac allograft rejection and vasculopathy in the cyclosporine-treated group was associated with prominent myocardial interferon-gamma gene expression, a finding absent in two thirds of the mycophenolate mofetil-treated swine. Moreover, the mycophenolate mofetil treated swine failed to develop IgM or IgG alloantibodies. CONCLUSIONS: A short course of mycophenolate mofetil resulted in a longer allograft survival than a similar course of cyclosporine. Moreover, mycophenolate mofetil reduced the prevalence of cardiac allograft vasculopathy as compared with cyclosporine treated controls. The salutary effect of mycophenolate mofetil may be related to its ability to decrease interferon-gamma expression in the myocardium and prevent the generation of alloantibodies. PMID- 16242458 TI - Invited commentary. PMID- 16242457 TI - Fibronectin-hepatocyte growth factor enhances reendothelialization in tissue engineered heart valve. AB - BACKGROUND: To overcome the limitations of tissue-engineered heart valves, which require cell seeding before implantation, a growth factor for in situ recellularization may be an important strategy. We developed a new decellularized valve containing a fusion protein combined fibronectin and hepatocyte growth factor. Here, we tested the hypothesis that our valve might accelerate in situ recellularization by inducing the proliferation of endothelial cells. METHODS: Porcine aortic valves were decellularized using detergent. Fibronectin-hepatocyte growth factor was introduced into the decellularized valves. The decellularized valves with fibronectin-hepatocyte growth factor were implanted into the pulmonary arterial trunk of dogs (F group: n = 15). As controls, decellularized valves without the growth factor (C group: n = 12), and with hepatocyte growth factor (H group: n = 12) were implanted in the same manner. Histologic examinations were performed 1 week and 1 month after implantation. RESULTS: One week after implantation, endothelial cells partially covered the surface of the graft in the F group but not the C and H groups. Although the C and H groups had inadequate recellularization 1 month after implantation, the F group showed a monolayer of endothelial cells, underneath which were areas of additional cell layers, which were vimentin positive. Quantitative evaluation demonstrated the amount of vimentin in the F group was 71% of the native control, and it was much lower in the other groups (C, 2.8%; H, 16.8%) 1 month after implantation. CONCLUSIONS: This study demonstrated that fibronectin-hepatocyte growth factor enhanced early in situ recellularization in decellularized valves. PMID- 16242459 TI - Hypoxia-reoxygenation, St. Thomas cardioplegic solution, and nicorandil on endothelium-derived hyperpolarizing factor in coronary microarteries. AB - BACKGROUND: We investigated effects of hypoxia-reoxygenation (H-R) with and without St. Thomas solution under clinically relevant temperatures and effects of nicorandil on endothelium-derived hyperpolarizing factor (EDHF)-mediated relaxation in porcine coronary microarteries. METHODS: In a myograph, rings of porcine microarteries (diameter 200 to 450 microm) were subjected to hypoxia (PO2 < 5 mm Hg) for 30 minutes in Krebs at 37 degrees C, or for 60 minutes in Krebs and St. Thomas solution with or without nicorandil (0.1 microM) at 37 degrees C or 4 degrees C, followed by 30-minute reoxygenation. The EDHF-mediated relaxation by bradykinin (-10 to approximately -6 logM) with inhibitors of nitric oxide and prostacyclin was studied. RESULTS: The maximal EDHF-mediated relaxation was reduced after hypoxia for 30 minutes (59.9%% +/- 1.6% versus 81.2%% +/- 3.5%, p < 0.05) or 60 minutes (44.4% +/- 6.0% versus 82.7% +/- 7.4%, p < 0.001) in Krebs or St. Thomas (28.9% +/- 1.8% versus 78.1% +/- 3.0%, p < 0.001) at 37 degrees C and at 4 degrees C (Krebs: 49.3% +/- 3.0%, p < 0.001; ST: 43.1% +/- 2.6%, p < 0.001) and it was less in St. Thomas solution at 37 degrees C than at 4 degrees C (p < 0.001). The reduced relaxation was recovered by nicorandil (Krebs at 37 degrees C: 81.7% +/- 3.4%, p < 0.001; St. Thomas at 37 degrees C: 71.0% +/- 7.9%, p <0.001; St. Thomas at 4 degrees C: 85.3% +/- 3.3%, p < 0.001). CONCLUSIONS: We conclude that (1) H-R impairs EDHF-mediated relaxation in the coronary microarteries with more injury during prolonged H-R, and this can be partially eliminated by St. Thomas at 4 degrees C but not at 37 degrees C; and (2) as an additive, nicorandil may fully restore EDHF-mediated endothelial function after prolonged H-R. PMID- 16242460 TI - Attenuation of DNA damage in canine hearts preserved by continuous hypothermic perfusion. AB - BACKGROUND: Continuous hypothermic perfusion is a novel cardiac preservation technique. Reactive oxygen species play a role in ischemia reperfusion injury and limit organ preservation. Oxidative stress mediates a DNA mismatch lesion (7, 8 dihydro-8-oxoguanine [8-oxo-G]), which is repaired by the enzymes MutY homologue (MYH), 8-oxo-G glycosylase (OGG1), and MutS homologue 2 (MSH2). We hypothesized that continuous hypothermic perfusion would allow for maintenance of cardiac function while attenuating myocardial DNA damage with respect to the current clinical practice of static preservation at 4 degrees C. METHODS: In our canine orthotopic transplant model, donor hearts were harvested after echocardiograms, and hemodynamic studies were obtained and served as controls. The hearts were transplanted after 24 hours of continuous hypothermic perfusion or 4 hours of static preservation, and were studied for 6 hours. Quantification of 8-oxo-G lesions, MYH, OGG1, and MSH2 concentrations were performed on biopsies using immunohistochemistry. RESULTS: Postimplant echocardiograms, completed in 7 continuously perfused and 8 statically preserved hearts, demonstrated good function and normal wall motion. Positive staining for 8-oxoG was markedly increased in the static preservation group. Staining density for MYH, OGG1, and MSH2 were significantly decreased in statically preserved hearts and equivalent between continuously perfused and control hearts. CONCLUSIONS: The DNA damage assayed by 8-oxoG was significantly increased in statically preserved versus continuously perfused hearts. The DNA repair enzymes MYH, OGG1, and MSH2 were also markedly decreased in the static preservation versus continuous hypothermic perfusion groups. Continuous hypothermic perfusion reduces oxidative damage and extends preservation without compromising function. PMID- 16242461 TI - Novel tissue-engineered biodegradable material for reconstruction of vascular wall. AB - BACKGROUND: To solve several problems with artificial grafts, we sought to develop a novel bioengineered material that can promote tissue regeneration without ex vivo cell seeding and that has sufficient durability to be used for artery reconstruction. Here, we tested whether this biodegradable material could accelerate the in situ regeneration of autologous cardiovascular tissue, especially of the arterial wall, in various models of cardiovascular surgeries. METHODS: The tissue-engineered patch was fabricated by compounding a collagen microsponge with a biodegradable polymeric scaffold composed of polyglycolic acid knitted mesh, reinforced on the outside with woven polylactic acid. Tissue engineered patches without precellularization were grafted into the porcine descending aorta (n = 5), the porcine pulmonary arterial trunk (n = 8), or the canine right ventricular outflow tract (as the large graft model; n = 4). Histologic and biochemical assessments were performed 1, 2, and 6 months after the implantation. RESULTS: There was no thrombus formation in any animal. Two months after grafting, all the grafts showed good in situ cellularization by hematoxylin/eosin and immunostaining. The quantification of the cell population by polymerase chain reaction showed a large number of endothelial and smooth muscle cells 2 months after implantation. In the large graft model, the architecture of the patch was similar to that of native tissue 6 months after implantation. CONCLUSIONS: A tissue-engineered patch made of our biodegradable polymer and collagen-microsponge provided good in situ regeneration at both the venous and arterial wall, suggesting that this patch can be used as a novel surgical material for the repair of the cardiovascular system. PMID- 16242462 TI - Invited commentary. PMID- 16242463 TI - An epidural cooling catheter protects the spinal cord against ischemic injury in pigs. AB - BACKGROUND: Using swine, we investigated whether epidural placement of a cooling catheter rather than infusing iced saline solution could protect the spinal cord from ischemia during aortic surgery. METHODS: We divided 14 domestic pigs into two groups of 7 each. Each underwent epidural catheter placement preceding 30 minutes of aortic cross-clamping distal to the origin of the left subclavian artery. In group 1, cold water was circulated continuously through the lumen of the catheter connected to an external unit. In group 2, animals received catheter placement without cooling. Spinal cord somatosensory evoked potentials were recorded. Neurologic status involving hind limbs was graded sequentially after surgery. RESULTS: At aortic cross-clamping, spinal temperature in group 1 (31.7 degrees +/- 0.6 degrees C) was significantly lower than in group 2 (37.8 degrees +/- 0.4 degrees C; p < 0.0001). No significant elevation of intrathecal pressure accompanied cooling with the catheter (group 1, 8.1 +/- 1.7 mm Hg; group 2, 8.0 +/- 1.5 mm Hg). Mean duration of total loss of potentials was significantly shorter in group 1 (7.4 +/- 3.8 minutes) than group 2 (19.7 +/- 7.3 minutes; p = 0.0002). Pigs in group 1 exhibited better hind limb function recovery (mean Tarlov score, 4.7 +/- 0.5) than group 2 (0.6 +/- 0.8; p = 0.0017). Group 1 showed normal histologic characteristics, whereas group 2 showed loss of motor neurons in the ventral horns. CONCLUSIONS: Epidural cooling catheter without iced saline infusion can cool the spinal cord without elevating intrathecal pressure, protecting the cord against ischemia. PMID- 16242464 TI - Invited commentary. PMID- 16242465 TI - Closure of the pericardium using synthetic bioabsorbable polymers. AB - BACKGROUND: Pericardial substitutes are known to ensure safer resternotomy at reoperation. A synthetic sheet made from expanded-polytetrafluoroethylene (e PTFE) has been most commonly used as a pericardial substitute. The e-PTFE sheet, however, can induce severe inflammatory reaction and diffuse fibrosis. This study was designed to investigate the absorption rate and tissue reaction associated with two absorbable pericardial substitutes: a gelatin sheet and L-lactic acid epsilon-caprolactone copolymer (L-C copolymer). In addition, e-PTFE sheet and autologous pericardium were used as controls. METHODS: Sixty dogs were divided into four groups of 15. In group A, a 3 x 3 cm segment of pericardium was excised, and the autologous pericardium was resutured. In group B, the pericardial defect was replaced with gelatin sheet. In group C, the defect was replaced with L-C copolymer sheet. In group D, the defect was replaced with e PTFE sheet. For each group, the implanted membranes were retrieved at 2 weeks (n = 1), 4 weeks (n = 3), 12 weeks (n = 5), and 24 weeks (n = 6) after implantation. RESULTS: The e-PTFE sheet produced severe adhesions to the heart and pleura and a more prominent inflammatory reaction, as compared with the gelatin sheet. The absorbable pericardial substitutes were completely absorbed by 24 weeks after implantation, and were replaced with fibrous membrane. CONCLUSIONS: Gelatin sheet may involve less adhesion and a reduced inflammatory reaction compared with e PTFE. PMID- 16242466 TI - Resection for tumors with carinal involvement: technical aspects, results, and prognostic factors. AB - BACKGROUND: Resection of tumors with carinal involvement remains a challenge because of specific problems of operative technique and airway management. We reviewed our experience with carinal resection and studied factors influencing postoperative course and long-term survival. METHODS: Between 1983 and 2002, 65 patients underwent a carinal resection for non-small-cell lung cancers involving the carina (54 squamous cell carcinomas and 11 adenocarcinomas). RESULTS: Fifty eight right sleeve pneumonectomies and 2 left sleeve pneumonectomies were performed. In addition, five tracheocarinal resections with double bronchial reimplantation (no lung resection) were also performed. The intraoperative airway management consisted of high-frequency jet ventilation in 83% of patients and intermittent conventional ventilation through the operative field in the remaining 17% of patients. Operative mortality was 7.7%. Resection was complete in 61 patients. The overall 5-year and 10-year survival rates were 26.5% and 10.6%, respectively. Patients with N0 or N1 disease had a 5-year survival of 38% compared with 5.3% for those with N2 disease (p < 0.01). At multivariate analysis only nodal status (N0, N1 versus N2; p = 0.0046) had a significant impact on long term survival. CONCLUSIONS: Carinal resection provides acceptable results in terms of operative mortality and long-term survival rates. Patients should be carefully selected and probably enrolled in a multimodality treatment program in case of anticipated mediastinal lymph node involvement. PMID- 16242467 TI - Pulmonary resection for metastatic malignant fibrous histiocytoma: an analysis of prognostic factors. AB - BACKGROUND: Factors affecting recurrence and survival after pulmonary resection for metastatic malignant fibrous histiocytoma (MFH) are not well known. METHODS: Records of patients undergoing pulmonary metastasectomy for MFH between January 1976 and January 2000 were analyzed. RESULTS: There were 103 patients (46 men and 57 women). Median age was 60 years (range, 20 to 86). Malignant fibrous histiocytoma metastasis was solitary in 29 patients (28%), multiple/unilateral in 33 (32%), and multiple/bilateral in 41 (40%). Median time interval from primary tumor resection to metastasectomy was 11 months (range, 0 to 86). Wedge resection was carried out in 87 patients (84%), segmentectomy in 3 (3%), lobectomy in 3 (3%), pneumonectomy in 2 (2%), and a combination of resections in 8 (8%). Resection was complete in 93 patients (90%). Complications occurred in 11 patients (11%) and included prolonged air leak in 7, and pneumonia, empyema, atelectasis, and sepsis in 1 each. One patient died (operative mortality, 1%). Follow-up ranged from 2 weeks to 153 months (median, 18 months). Five-year survival was 21% (95% confidence interval, 14% to 31%). In the 103 patients, incomplete resection (p < 0.0001) was associated with decreased survival. Among the 93 patients with complete resection, factors associated with decreased survival included the presence of extrapulmonary disease at time of metastasectomy (p = 0.01), more than two nodules resected (p = 0.001), and adjuvant therapy after metastasectomy (p = 0.0007). CONCLUSIONS: Pulmonary resection of metastatic MFH is safe. Improved survival was associated with the absence of extrapulmonary disease at time of metastasectomy, with fewer than three pulmonary nodules resected, and with a complete resection. PMID- 16242468 TI - Quantitative computed tomography versus spirometry in predicting air leak duration after major lung resection for cancer. AB - BACKGROUND: Emphysema is a well-known risk factor for developing air leak or persistent air leak after pulmonary resection. Although quantitative computed tomography (CT) and spirometry are used to diagnose emphysema, it remains controversial whether these tests are predictive of the duration of postoperative air leak. METHODS: Sixty-two consecutive patients who were scheduled to undergo major lung resection for cancer were enrolled in this prospective study to define the best predictor of postoperative air leak duration. Preoperative factors analyzed included spirometric variables and area of emphysema (proportion of the low-attenuation area) that was quantified in a three-dimensional CT lung model. Chest tubes were removed the day after disappearance of the air leak, regardless of pleural drainage. Univariate and multivariate proportional hazards analyses were used to determine the influence of preoperative factors on chest tube time (air leak duration). RESULTS: By univariate analysis, site of resection (upper, lower), forced expiratory volume in 1 second, predicted postoperative forced expiratory volume in 1 second, and area of emphysema (< 1%, 1% to 10%, > 10%) were significant predictors of air leak duration. By multivariate analysis, site of resection and area of emphysema were the best independent determinants of air leak duration. The results were similar for patients with a smoking history (n = 40), but neither forced expiratory volume in 1 second nor predicted postoperative forced expiratory volume in 1 second were predictive of air leak duration. CONCLUSIONS: Quantitative CT is superior to spirometry in predicting air leak duration after major lung resection for cancer. Quantitative CT may aid in the identification of patients, particularly among those with a smoking history, requiring additional preventive procedures against air leak. PMID- 16242469 TI - Spontaneous hemopneumothorax revisited: clinical approach and systemic review of the literature. AB - BACKGROUND: Spontaneous hemopneumothorax, defined as the accumulation of more than 400 mL of blood in the pleural cavity in association with spontaneous pneumothorax, is a rare entity occurring in young patients and may be life threatening. Although many reports of case studies and series have been published in the world literature, the lack of consistent intraoperative findings and varying surgical methods require a review study. METHODS: We discuss the clinical features, management, surgical findings, and outcomes of our own patients with spontaneous hemopneumothorax. RESULTS: From September 1997 to September 2003, 488 patients with spontaneous pneumothorax were treated at our hospital. Of these patients, 27 (5.5%) had spontaneous hemopneumothorax develop. These 27 patients were comprised of 25 men and 2 women ranging in age from 15 to 39 years (mean age, 22.3 years). The amount of blood that was drained ranged from 400 to 1,700 mL (mean, 1,012 mL). Twenty-one patients underwent video-assisted thoracoscopic surgery within 1 day after admission; the remaining 6 patients were treated conservatively with tube thoracostomy alone. On arrival at our emergency room, 9 patients (33.3%) experienced hemodynamic instability with hypovolemic shock. In a review of 6,396 patients with spontaneous pneumothorax in the literature and in our current study, 201 patients (3.1%) had spontaneous hemopneumothorax develop. One hundred seventy-six patients (87.6%) were treated surgically, whereas video assisted thoracoscopic surgery has been performed in 48.9% of patients (86 of 176). There was no recurrence of hemopneumothorax in any of the 201 patients with spontaneous hemopneumothorax after treatment during the follow-up period. CONCLUSIONS: Thus one-third of the patients with spontaneous hemopneumothorax had shock symptoms develop. Video-assisted thoracoscopic surgery may be considered as an initial treatment procedure for patients with spontaneous hemopneumothorax, whereas conservative treatment is effective and may be performed in selected patients. PMID- 16242470 TI - Postoperative chylothorax after cardiothoracic surgery in children. AB - BACKGROUND: The purpose of this study is to determine the incidence, risk factors, and outcomes for chylothorax in children undergoing cardiothoracic surgery. METHODS: Hospital databases were used to identify chylothorax cases. Surgical databases were used to identify all patients undergoing cardiothoracic surgery. Medical records were reviewed, including daily records of drainage volumes and management. RESULTS: From September 2000 to December 2002, there were 48 cases of chylothorax in 1,257 surgeries--an incidence of 3.8% (95% confidence interval: 2.8% to 4.8%). Overall mortality rate was similar, but cases had longer postoperative hospital stays (median, 22 versus 8 days; p < 0.001). Incidence of chylothorax was significantly higher with heart transplantation and Fontan procedures. Diagnosis was made at a median of 6 days after surgery. Duration of drainage was a median of 15 days, with 11 patients draining more than 30 days. Longer duration of drainage was associated with cavopulmonary anastomosis procedures and longer time to diagnosis of chylothorax. Nutritional management included low fat diet, enteral feeds enriched with medium-chain triglycerides, and parenteral nutrition. Five patients were treated with octreotide, 4 with thoracic duct ligation, and 1 with pleurodesis. Octreotide was associated with a variable effect on drainage. Thoracic duct ligation reduced, but did not stop drainage. CONCLUSIONS: Chylothorax increases duration of hospitalization after cardiovascular surgery in children. Early diagnosis may reduce the duration of chylothorax. Nutritional strategies remain the cornerstone for management of postoperative chylothorax. The impact of octreotide and surgical intervention is limited when reserved for patients with severe or prolonged drainage. PMID- 16242471 TI - Invited commentary. PMID- 16242472 TI - Improved results treating lung allograft failure with venovenous extracorporeal membrane oxygenation. AB - BACKGROUND: Primary graft failure remains a significant source of mortality after lung transplantation. Extracorporeal membrane oxygenation (ECMO) provides treatment for affected recipients. We hypothesized that venovenous membrane oxygenation provides a safer alternative than venoarterial support for lung recipients suffering from primary graft failure. METHODS: We conducted an analysis of 522 patients who underwent lung transplantation from April 1992 to July 2004. Twenty-three (4.5%) patients required membrane oxygenation secondary to primary graft failure unresponsive to conventional treatment. Of these recipients, 15 (65%) were treated with venoarterial, while 8 (35%) underwent venovenous membrane oxygenation. RESULTS: Median days to initiation and duration of membrane oxygenation did not differ between groups. Eight of 15 patients (53%) from the venoarterial group were successfully weaned from life support, with one surviving greater than 45 days. This lone long-term survivor required retransplantation 4 days after initial transplant. In contrast, all venovenous patients were weaned from support, with 7 of 8 surviving greater than 30 days. The 30-day survival for venovenous recipients (88%) approximates that of all lung recipients at our center (94%, p = 0.42). Noted complications for ECMO patients included renal failure (n = 16), neurologic catastrophes (n = 8), sepsis (n = 5), and hemorrhage (n = 10). The venoarterial recipients suffered 30 of 39 total complications. Most of the complications for venovenous recipients involved renal failure, but by hospital discharge these patients demonstrated a mean creatinine of 0.9 mg/dL. CONCLUSIONS: For lung recipients with primary graft failure, venovenous membrane oxygenation provides better outcomes, with fewer complications, than venoarterial membrane oxygenation. PMID- 16242473 TI - A strategy for determining which thoracic esophageal cancer patients should undergo cervical lymph node dissection. AB - BACKGROUND: There is controversy about performing cervical lymph node dissections in all middle and lower thoracic esophageal squamous cell carcinoma patients. The purpose of this study was to evaluate whether intraoperative examination of thoracic paratracheal lymph node by real-time reverse transcription-polymerase chain reaction was worthwhile for selecting patients for cervical lymph node dissection. METHODS: Under informed consent, 30 middle and lower thoracic esophageal squamous cell carcinoma patients were examined for thoracic paratracheal lymph node metastasis intraoperatively by hematoxylin-eosin staining and real-time reverse transcription-polymerase chain reaction for messenger RNA encoding squamous cell carcinoma antigen. When thoracic paratracheal lymph node metastasis was found, cervical lymph node dissection was performed. If the patients had no thoracic paratracheal lymph node metastasis, a randomized study for selection of cervical lymph node dissection was performed. RESULTS: Eleven of 30 patients with middle or lower third thoracic esophageal squamous cell carcinoma had thoracic paratracheal lymph node metastasis. Five of these 11 patients had cervical lymph node metastasis. Nineteen patients who had no metastasis in the thoracic paratracheal lymph nodes were enrolled in a randomized study. Eight of the 19 patients received cervical lymph node dissection and they were found not to have cervical lymph node metastasis. The other 11 patients did not receive cervical lymph node dissection, and there was no cervical lymph node recurrence. CONCLUSIONS: The intraoperative diagnosis of metastasis in the thoracic paratracheal lymph node may be used as an indicator for cervical lymph node dissection in middle and lower thoracic esophageal cancer patients. PMID- 16242474 TI - LIFEBRIDGE: a portable, modular, rapidly available "plug-and-play" mechanical circulatory support system. AB - PURPOSE: We describe the LIFEBRIDGE, a portable, modular, rapidly available "plug and-play" mechanical circulatory support system, and report its experimental safety evaluation. DESCRIPTION: The modular construction consists of a disposable patient module with cardiopulmonary bypass circuit, control module, and base module with power supply, embedded PC, and user interface. The system weighs about 20 kg, has a modular design, and has semi-automatic priming that allows action within 5 minutes, and a 7-step air elimination program that prevents air embolization. EVALUATION: In eight pigs (85 +/- 10 kg) we investigated this system using central (right atrium and ascending aorta) cannulation (n = 4) or peripheral (iliac) cannulation (n = 4). Pump flows were 5.7 +/- 0.2 L/min with central and 4.1 +/- 0.2 L/min with peripheral cannulation, yielding sufficient animal perfusion and gas exchange. Using an intraaortic 8-MHz Doppler device, we demonstrated that venous air boluses of up to 100 mL were effectively removed, thus avoiding air embolization. Changing heights between animals and LIFEBRIDGE did not affect its proper action. CONCLUSIONS: This initial evaluation demonstrates that the LIFEBRIDGE is rapidly available, provides adequate perfusion and gas exchange, and operates safely even under simulated transport conditions. PMID- 16242475 TI - Invited commentary. PMID- 16242476 TI - Off-pump CABG with synchronized arterial flow ensuring system. AB - PURPOSE: We developed a synchronized, arterial-flow, ensuring system to perform coronary anastomoses safely without any ischemia-related event. DESCRIPTION: Arterial blood is removed from the femoral artery. The resulting blood passes a switching valve and is pumped out to a syringe pump. This pump controller provides pulsatile arterial blood flow synchronized with the diastolic phase on an electrocardiogram. The arterial blood is perfused to the coronary artery through a fine flexible cannula during anastomosis. EVALUATION: From February 1999, 524 consecutive patients were operated on using the synchronized arterial flow ensuring system. Mean duration for each anastomosis was 7.6 +/- 3.3 minutes (range, 4 to 20 min). There were no intraoperative fatal arrhythmias, ventricular arrhythmias, or short-run or hemodynamic deterioration during anastomoses. No hospital death was observed, and postoperative myocardial infarction occurred in 2 patients (0.4%). Postoperative angiography showed a 98.1% patency rate. CONCLUSIONS: The early clinical and angiographical results for off-pump CABG with the synchronized arterial flow ensuring system were excellent without mortality. We believe that off-pump CABG can be more safely performed using the synchronized arterial flow ensuring system based on our favorable results. PMID- 16242477 TI - Computer-controlled stapling system for lung surgery. AB - PURPOSE: Current disposable hand-actuated staplers may pose reliability problems, especially with respect to the measurement of tissue thickness. We have evaluated a newly developed stapler with a computer-controlled placement of staples. DESCRIPTION: The SurgAssist system (Power Medical Interventions, New Hope, PA) is comprised of a console that houses a computer, a remote control unit, a flexible shaft, and a cartridge. The remote control unit has two uses: (1) controlling the accurate placement of the cartridge by orientating the tip of the flexible shaft, and (2) controlling the closure of the stapler and the firing. Each cartridge contains a programmed electronic device that triggers the activation of the appropriate program in the main microprocessor. The compression level on the tissue is determined by the computer. EVALUATION: The system was used in a consecutive series of 38 patients, 26 times during open lung surgery and 12 times during video-assisted thoracic surgery. The following open procedures were performed: three pneumonectomies, 15 lobectomies, three segmentectomies, and five wedge resections. The following video-assisted thoracic surgery procedures were performed: eight wedge resections and four bullectomies for pneumothorax. There was no stapling failure and no complication related to the use of the stapler. During video-assisted thoracic surgery, some ergonomic problems were encountered that will be overcome by redesign. CONCLUSIONS: The computer-controlled stapling system may significantly improve tissue approximation during open and video assisted thoracic surgery. PMID- 16242478 TI - Invited commentary. PMID- 16242479 TI - Inflammatory reaction after sirolimus-eluting stent implant. AB - Sirolimus-eluting stents (SES) are supposed to attenuate cell proliferation and reduce restenosis rate. Histologic finding from coronary artery after SES implant showed fibrosis and inflammatory infiltrate, revealing a chronic inflammatory reaction. Extension of coronary inflammatory reaction after stenting needs clarification. The long-term consequences are unknown. PMID- 16242480 TI - Respiratory dependent compression of a venous bypass: therapy by stenting. AB - Although coronary artery bypass surgery has provided major advances in the treatment of coronary artery disease, narrowing of bypass vessels still constitutes a drawback of this therapy. Although this event is most frequently caused by intraluminal processes, obstruction from external structures is extremely rare. We report such a case in which external bypass compression was provoked by deep inspiration causing typical anginal symptoms. Percutaneous coronary intervention including stent placement provided bypass patency independent from the patient's respiratory phase. Disappearance of symptoms and absence of myocardial ischemia in perfusion scans confirmed successful treatment. PMID- 16242481 TI - Transcatheter repair of recurrent postinfarct ventricular septal defects. AB - Surgical repair of recurrent postmyocardial infarction septal defect is associated with a high mortality rate. We present 2 patients whose recurrent defects were closed percutaneously using an Amplatzer device. PMID- 16242482 TI - Chronic mitral valve rejection requiring replacement in a nine-year-old allograft. AB - A 43-year-old woman underwent mitral valve replacement for severe mitral regurgitation nine years after orthotopic heart transplant. Histopathology showed chronic rejection of the mitral valve with lymphocytic infiltrates. The patient is well at one year follow-up. This report describes an identified case of chronic mitral valve rejection requiring valve replacement. PMID- 16242483 TI - Mitral valve replacement for mitral stenosis secondary to Hunter's syndrome. AB - Hunter's syndrome is a rare, X-linked recessive, mucopolysaccharidosis. Survival into adulthood is uncommon. Mitral valve disease, predominantly regurgitation, has been reported in these patients. We have found no reports of mitral valve replacement for mitral stenosis secondary to Hunter's syndrome in the English literature. We report that mitral valve replacement for this pathology is a viable treatment option in an adult patient; however, specific precautions must be considered. PMID- 16242484 TI - Idiopathic inflammatory aneurysm of the ascending aorta. AB - Inflammatory aneurysms of the ascending aorta are exceedingly rare. The principal cause of ascending aortic aneurysms remains arteriosclerosis. We present a patient with an ascending aortic aneurysm caused by active inflammatory disease superimposed on complex atherosclerotic disease of the aorta. PMID- 16242486 TI - Anomalous origin of the right pulmonary artery from the ascending aorta with fibrous continuity to the pulmonary trunk. AB - A 1-month-old patient was brought to our institution with clinical signs of pulmonary hypertension. Cross-sectional echocardiography suggested a diagnosis of aortopulmonary window. At the time of surgery, we found that the right pulmonary artery was arising anomalously from the left side of the ascending aorta, but was also connected to the pulmonary trunk by a fibrous cord. We reimplanted the right pulmonary artery into the pulmonary trunk, closing the resultant opening in the ascending aorta by direct suture. The postoperative course was uneventful. On follow-up, the patient is asymptomatic without medication. PMID- 16242485 TI - Successful surgical treatment of Takayasu's arteritis associated with pyoderma gangrenosum. AB - Pyoderma gangrenosum is rarely associated with Takayasu's arteritis. We report the successful surgical treatment of a 34-year-old woman who was diagnosed with pyoderma gangrenosum associated with thoracic aortic aneurysm and dissection due to Takayasu's arteritis. She underwent graft replacement of the thoracic aortic aneurysm under cardiopulmonary bypass, with perioperative management using prednisolone and cyclosporine A. She has had no evidence of the development of skin lesions or the progression of Takayasu's arteritis. PMID- 16242487 TI - Circumflex coronary artery from right pulmonary artery in hypoplastic left heart syndrome. AB - We report the case of a newborn with the very rare association of hypoplastic left heart syndrome and aberrant origin of the circumflex coronary artery from the right pulmonary artery. This condition can jeopardize the result of the Norwood palliation. PMID- 16242488 TI - Successful use of Thoratec biventricular support in a small child awaiting cardiac transplantation. AB - The use of paracorporeal mechanical biventricular support devices either as a bridge to transplantation or while awaiting recovery of the failing heart has been well described in the literature. The majority of these reports detail conditions specific to the adult population. We describe use of the Thoratec ventricular assist device (Thoratec Corp, Pleasanton, CA) in the smallest known cardiomyopathy patient to date to be successfully supported with an emergent biventricular device before subsequent cardiac transplantation. The operative technique, pump settings, and modifications utilized to optimize peripheral perfusion are detailed. PMID- 16242489 TI - Annuloplasty of the regurgitant mitral valve after myocarditis in children. AB - Acute myocarditis is characterized by the development of rapid life-threatening congestive heart failure and arrhythmias. In many cases with hemodynamic compromise, medical therapy and mechanical support alone are not sufficient. Various surgical procedures have been tried to bridge patients with myocarditis to both transplant and recovery. Mitral regurgitation is a frequent association with end stage cardiomyopathy and predicts poor outcome. Mitral annuloplasty is well-established in adults with ischemic and dilated cardiomyopathy and the results are superior to medical therapy alone and are comparable to cardiac transplantation. However, its effectiveness and use is not well-established in children with cardiomyopathy. We report our experience in two children. PMID- 16242490 TI - Proximal conduit obstruction after Sano modified Norwood procedure. AB - Sano and colleagues recently described a modification of first stage palliation for hypoplastic left heart syndrome utilizing a right ventricle to pulmonary artery conduit. Preliminary results are favorable, but experience with this technique is limited. We report a case of sudden death due to obstruction of the proximal conduit by fibrointimal hyperplasia. This case of lethal conduit obstruction presented 3 months after initial palliation. Early cardiac catheterization and second stage palliation may be necessary to minimize the risk of such adverse events after the Sano modification. PMID- 16242491 TI - Use of a modified Dumon stent for postoperative bronchopleural fistula. AB - This report describes a case of a postoperative bronchopleural fistula successfully managed with a modified Dumon stent. Surgical interventions for the bronchopleural fistula with empyema were subsequently avoided. Dumon stent is an acceptable option for the treatment of postoperative bronchopleural fistulas. PMID- 16242492 TI - Occlusion of a broncho-cutaneous fistula with endobronchial one-way valves. AB - Bronchopleural and broncho-cutaneous fistulas can be problematic after lobectomy for tumors or aspergillomas. Closure of the air leak and treatment of infection are essential to allow the fistula to heal. The initial treatment can usually proceed along standard lines, but if the fistula persists, then treatment can be problematic. This report is the first description of the use of multiple Emphasys Medical endobronchial valve prostheses (Emphasys Medical, Inc, CA) to control a previously intractable broncho-cutaneous fistula. The valves have been specifically designed for airway placement as part of a therapeutic approach to severe emphysema. The advantages of using valves in this situation are discussed. PMID- 16242493 TI - New treatment method for reexpansion pulmonary edema: differential lung ventilation. AB - Reexpansion pulmonary edema (REPE) is an iatrogenic complication that develops in a lung rapidly reinflated after varied periods of collapse. Though infrequent, it is often lethal. Measures to prevent REPE are well known and mild cases may be reversed with conventional ventilator therapy and medication; however, there is no definitive treatment for this condition in its severest forms. Asynchronous differential lung ventilation is a relatively new therapeutic modality for preoperative, intraoperative, or postoperative treatment of respiratory failure secondary to ventilation-perfusion mismatch. Successful treatment of severe REPE in a 29-year-old male patient is described, using asynchronous differential lung ventilation. PMID- 16242494 TI - Intrathoracic splenosis. AB - Intrathoracic splenosis is a rare condition involving autotransplantation of the splenic tissue into the pleural cavity. It is typically a result of diaphragmatic and splenic rupture after blunt or penetrating abdominal trauma. The diagnosis is usually determined by surgical biopsy of an incidental asymptomatic mass on a radiograph or computed tomography. The process is benign and can be generally followed with serial chest radiographs. We herein discuss a 34-year-old male who presented with sharp chest pains. PMID- 16242495 TI - Tracheobronchopathia osteochondroplastica occurring in a subsegmental bronchus and causing obstructive pneumonia. AB - A 59-year-old man was referred to the University of Occupational and Environmental Health hospital because a chest roentgenogram obtained at an annual medical examination showed an abnormal shadow. We treated the patient for tracheobronchopathia osteochondroplastica, which caused segmental obstructive pneumonia that proved to be difficult to accurately diagnose before surgery. PMID- 16242496 TI - Antireflux operation for gastroesophageal reflux after Roux-en-y gastric bypass for obesity. AB - Gastroesophageal reflux disease (GERD) affects many morbidly obese patients. The Roux-en-y gastric bypass operation often achieves the simultaneous aims of weight reduction as well as reflux correction. However, some patients continue to suffer from severe gastroesophageal symptoms after successful postoperative weight reduction. There are no reports describing surgical treatments for these patients. Here we report our management of intractable postoperative reflux with a Belsey Mark IV fundoplication performed one year after a successful Roux-en-y gastric bypass operation. The patient tolerated the operation without problems and experienced successful resolution of her reflux symptoms. PMID- 16242497 TI - Images in cardiothoracic surgery. Active malignant pleural effusion captured through the thoracoscope. PMID- 16242498 TI - Images in cardiothoracic surgery. Herniation of postpneumonectomy space fluid. PMID- 16242499 TI - Pulmonary root translocation for repair of Taussig-Bing anomaly with interrupted arch. AB - Anterior translocation of the pulmonary root was used as a new approach to the staged repair of Taussig-Bing anomaly with an interrupted aortic arch. It was performed to construct the right ventricle outflow tract with intraventricular baffling of the left ventricle to the aorta as the second stage operation after repair of the interrupted arch and pulmonary artery banding. This technique allows minimization of pulmonary regurgitation and has the major theoretical advantage for growth potential, which could diminish the need for reoperation. PMID- 16242500 TI - A simple system to deliver blood cardioplegia. AB - We describe a simple and inexpensive system designed to deliver blood cardioplegia either diluted or at the patient's hematocrit, with controlled temperature and additive concentration. This system can be applied to any pump set, and suits any strategy for clinical myocardial preservation. PMID- 16242501 TI - Multiple arterial revascularization using the tangential K-graft technique. AB - The tangential K graft is a comfortable surgical technique aiming to increase cardiac surgeons' versatility in performing multiple arterial grafting using only two arterial conduits. One end of the free graft--either the right internal thoracic artery (RITA) or the radial artery (RA)--is attached to a marginal circumflex branch. Its other end is anastomosed end to side to a diagonal branch. After the left internal thoracic artery (LITA) is attached to the left anterior descending coronary artery, a wide-open side-to-side LITA to free RITA or RA anastomosis--resembling the letter K--is constructed. PMID- 16242502 TI - A novel surgical correction through a small transverse incision for pectus excavatum. AB - Surgical correction of pectus excavatum (PE) has been well established since Ravitch's publication in 1949. However, Ravitch's procedure, even if modified, was associated with the relatively radical nature of the operation. The aim of this study was to report our early experience and results in treatment of PE by a novel less invasive surgical technique through a small skin incision. From 1998 to 2003, a novel surgical correction through a small transverse incision was performed for 11 patients with PE, including 9 males and 2 females. The mean age was 9.2 years (range, 3 to 17 years). The less invasive surgical technique consisted of a small transverse skin incision over the deepest part of the PE deformity, subcutaneous dissection to the margin of the depressed deformity, elevation of pectoralis musculature from the midline toward the lateral border of the operative field, subperichondrial resection of the short segment (1 to 2 cm) of the involved costal cartilages, detachment of the xiphoid process and elevation of the sternum with sharp or blunt dissection, retrosternal titanium miniplate strutting, placement of drainage tubes in the mediastinum or pleural spaces, and closure of the operative wound. No sternal osteotomy was performed in this series. The average length of the skin incision was 3.2 cm. The number of the resected cartilages varied from 3 to 6 ribs on each side. The average blood loss was 41 mL (range, 10 to 80 mL), and the operation time was 3.1 hours. The duration of hospitalization was 4.4 days on average. There was no surgical complication or mortality. All patients were satisfied with their cosmesis, and no migration of the retrosternal strut was found in chest radiographs until the date of analysis. This less invasive surgical technique, which did not require osteotomy, could be effectively performed through a small skin incision and was associated with steady recovery of chest wall deformity, as well as excellent cosmetic results. PMID- 16242503 TI - Neonatal brain protection and deep hypothermic circulatory arrest: pathophysiology of ischemic neuronal injury and protective strategies. AB - Deep hypothermic circulatory arrest (DHCA) has been used for the past 50 years in the surgical repair of complex congenital cardiac malformations and operations involving the aortic arch; it enables the surgeon to achieve precise anatomical reconstructions by creating a bloodless operative field. Nevertheless, DHCA has been associated with immediate and late neurodevelopmental morbidities. This review provides an overview of the pathophysiology of neonatal hypoxic brain injury after DHCA, focusing on cellular mechanisms of necrosis, apoptosis, and glutamate excitotoxicity. Techniques and strategies in neonatal brain protection include hypothermia, acid base blood gas management during cooling, and pharmacologic interventions such as the use of volatile anesthetics. Surgical techniques consist of intermittent cerebral perfusion during periods of circulatory arrest and continuous regional brain perfusion. PMID- 16242504 TI - Acquiring proficiency in off-pump surgery: traversing the learning curve, reproducibility, and quality control. AB - As the risk profile of patients considered for surgical revascularization worsens, the cumulative benefit of off-pump coronary artery bypass (OPCAB) over conventional coronary artery bypass grafting, in terms of lower morbidity and reduced healthcare costs, may increase. There is still resistance to the introduction of OPCAB surgery however, its practice is variable and surgical residents are rarely trained in these techniques. This article considers how the learning curve in OPCAB may be negotiated and prospectively monitored to ensure quality control. The evidence suggests that situations in which suitable senior expertise exists, OPCAB surgery can be introduced into surgical practice and safely taught to trainees without detriment to patients. This is achieved by a progressive increase in the complexity of the case mix and careful early supervision. The introduction of OPCAB has coincided with the increasing use of control charts as quality control tools. Performance monitoring provides reassurance that patients are not being put at risk during the introduction of OPCAB; control chart methods can be used prospectively for real time performance monitoring by consultant surgeons and residents alike. These techniques may ultimately be used to determine proficiency and accreditation. Increasing use of parallel training techniques, the development of structured training programs that encompass OPCAB and other new technologies in cardiac surgery, coupled with objective performance monitoring are warranted to meet the needs of a changing patient population. PMID- 16242505 TI - Albert Wojciech Adamkiewicz: the discoverer of the variable vascularity of the spinal cord. AB - The present article contains a brief biography as well as a discussion of the more significant contributions of Albert Wojciech Adamkiewicz, Head of General and Experimental Pathology at the Jagiellonian University in Cracow in the years 1879 to 1892, and who is now primarily remembered as the discoverer of the major radicular artery eponymically named "Adamkiewicz's artery." His work as an investigator of the variability of the vasculature of the spinal cord led to his major contributions to present clinical practice in such areas as vascular surgery, neurosurgery, pediatric surgery, and the surgery of the aorta, providing a permanent Polish eponymic accent in major textbooks in these specialties. This article also deals with Adamkiewicz's contributions in other fields, mostly involving the nervous system, such as the development of a new and original method for staining neuronal tissue (double staining of the spinal cord) and his extensive studies of spinal cord degeneration. The authors also present aspects of his career that brought ill fame to Adamkiewicz. These were his claims to have discovered the so-called nervous bodies and anticancer antitoxin, which were both severely criticized by his faculty peers at the Jagiellonian University. The biography is supplemented with an attempt at evaluating Adamkiewicz's entire scientific output, wherein unquestionable achievements and pointed discoveries prevail in comparison with failures. PMID- 16242506 TI - PDA ligation: trans or extrapleural approach. PMID- 16242508 TI - Exhaled nitric oxide after cardiopulmonary bypass. PMID- 16242510 TI - How should bronchioloalveolar carcinoma of the lung 3 centimeters or less be treated? PMID- 16242512 TI - Relearning the lessons of the past: are we making progress? PMID- 16242514 TI - Loculated neonatal chylothorax treated with octreotide: complete recovery while on unrestricted full fat breast milk. PMID- 16242515 TI - Transxiphoid hand-assisted thoracoscopic surgery: an approach included in the armamentarium. PMID- 16242519 TI - What is nanomedicine? PMID- 16242520 TI - Nanostructure-mediated drug delivery. PMID- 16242521 TI - Formation of nanoparticles of a hydrophilic drug using supercritical carbon dioxide and microencapsulation for sustained release. PMID- 16242522 TI - Atomic force microscopy analysis of the Huntington protein nanofibril formation. PMID- 16242524 TI - Levosimendan improves postresuscitation outcomes in a rat model of CPR. AB - In this study we sought to determine whether a calcium sensitizer, levosimendan, would have a more favorable effect on postresuscitation myocardial function and, consequently, postresuscitation survival than beta-adrenergic dobutamine. The extreme decrease in survival before hospital discharge of resuscitated victims is attributed, in part, to postresuscitation myocardial failure, and dobutamine has been recommended for the management of postresuscitation myocardial failure. We studied a total of 15 animals. Ventricular fibrillation was induced in Sprague Dawley rats weighing 450 to 550 g. Cardiopulmonary resuscitation (CPR), including chest compressions and mechanical ventilation, was begun after 8 minutes of untreated cardiac arrest. Electrical defibrillation was attempted after 6 minutes of CPR. Each animal was resuscitated. Animals were randomized to undergo treatment with levosimendan, dobutamine, or saline-solution placebo. These agents were administered 10 minutes after the return of spontaneous circulation. Levosimendan was administered in a loading dose of 12 microg kg(-1) over a 10 minute period, followed by infusion of 0.3 microg kg(-1) min(-1) over the next 230 minutes. Dobutamine was continuously infused at a dosage of 3 microg kg(-1) min(-1). Saline-solution placebo was administered in the same volume and over the same amount of time as levosimendan. Levosimendan and dobutamine produced comparable increases in cardiac output and rate of left-ventricular pressure increase. However, administration of levosimendan resulted in lower heart rates and lesser increases in left ventricular diastolic pressure compared with both dobutamine and placebo. The duration of postresuscitation survival was significantly greater with levosimendan (16 +/- 2 hours), intermediate with dobutamine (11 +/- 2 hours) and least with saline-solution placebo (8 +/- 1 hour). Levosimendan and dobutamine both improved postresuscitation myocardial function. However, levosimendan produced more favorable postresuscitation myocardial function and increased the duration of postresuscitation survival. PMID- 16242525 TI - Heterogeneity of cag genotypes and clinical outcome of Helicobacter pylori infection. AB - Helicobacter pylori infecting strains may include colony subtypes with different cytotoxin-associated gene (cag) genotypes. We sought to determine whether the cag heterogeneity of infecting strains is related to the clinical outcome of infection. Gastric biopsies for culture and histologic study were taken from 19 patients infected with cagA-positive strains (6 with duodenal ulcer, 8 with atrophic gastritis, and 5 with nonatrophic gastritis). For each biopsy, DNA was extracted from 10 single colonies and from a sweep of colonies. Polymerase chain reaction (PCR) for cagA and cagE (both located in the right half of cag) and virB11 (located in the left half of cag) was performed. Random amplified polymorphic DNA PCR (RAPD-PCR) and sequencing of glmM PCR product were performed to verify strain identity of colonies with different cag genotypes. In all patients, PCR from sweeps were positive for cagA, showing that all specimens contained cagA-positive H. pylori subtypes. In 11 patients, PCR products from all colonies were positive for cagA, cagE, and virB11, but in 8 patients, PCR products from varying numbers of colonies were negative for 1 or more cag genes. RAPD-PCR and sequencing of glmM PCR product confirmed the strain identities of colonies with different cag genotypes. We detected cag deletions in 6 of 8, 2 of 5, and 0 of 6 patients with atrophic gastritis, nonatrophic gastritis, and duodenal ulcer, respectively (P = .02). In conclusion, changes in cag genotype in single colony isolates from subjects infected with cagA-positive H. pylori strains are more common in atrophic than in nonatrophic gastritis or duodenal ulcer. These findings are consistent with host-induced (acid secretion?) adaptive changes in cag genotype during infection. PMID- 16242526 TI - Characterization and gene transfer in mesenchymal stem cells derived from human umbilical-cord blood. AB - It has been shown that the stromal-cell population found in bone marrow can be expanded and differentiated into cells with the phenotypes of bone, cartilage, muscle, neural, and fat cells. However, whether mesenchymal stem cells (MSCs) are present in human umbilical-cord blood (UCB) has been the subject of ongoing debate. In this study, we report on a population of fibroblastlike cells derived from the mononuclear fraction of human UCB with osteogenic and adipogenic potential, as well as the presence of a subset of cells that have been maintained in continuous culture for more than 6 months. These cells were found to express CD29, CD44, CD90, CD95, CD105, CD166, and MHC class, but not CD14, CD34, CD40, CD45, CD80, CD86, CD117, CD152, or MHC class II. We also compared gene expression after gene transfer using lenti- and adenoviral vectors carrying the green fluorescence protein to the MSCs derived from UCB because a reliable gene delivery system is required to transfer target genes into MSCs, which have attracted attention as potential platforms for the systemic delivery of therapeutic genes. The lentiviral vectors can transduce these cells more efficiently than can adenoviral vectors, and we maintained transgene expression for at least 5 weeks. This is the first report showing that UCB-derived MSCs can express exogenous genes by way of a lentivirus vector. These results demonstrate that human UCB is a source of mesenchymal progenitors and may be used in cell transplantation and a wide range of gene-therapy treatments. PMID- 16242527 TI - Serum complement factor I decreases Staphylococcus aureus phagocytosis. AB - Complement-mediated opsonization of Staphylococcus aureus is a critical host defense in animal models. Specifically, C3b and CD35 play important roles in effective opsonophagocytosis of S. aureus. We have shown that complement control protein factor I mediates cleavage of the complement opsonin C3b bound to the S. aureus surface. In this study, we examined the physiologic relevance of this observation by determining whether factor I-mediated cleavage of S. aureus-bound C3b decreased phagocytosis of S. aureus by neutrophils. Compared with controls, anti-factor I antibody inhibited C3b-cleavage on the S. aureus surface by >83% (as measured by iC3b generation) and increased phagocytosis of S. aureus by >100%. Treatment of C3b-coated S. aureus with factor I increased generation of iC3b (75%), decreased the total amount of C3-fragments bound to the S. aureus surface (58%), and decreased the number of bacteria phagocytosed (40%). Testing specifically for C3-fragments shed from the S. aureus surface, we found that factor I increased shedding (43%). Notably, these factor I-mediated effects were of the same magnitude regardless of whether factor H, a known cofactor for factor I, was present. These findings indicate that S. aureus benefits from, and possibly manipulates, the normally host-protective activity of factor I cleavage of C3b, which results in bacterial escape from complement-mediated opsonophagocytosis. Because escaping opsonophagocytosis-mediated destruction is a necessary mechanism for bacterial survival resulting in human disease, preventing cleavage of C3b on the S. aureus surface, and thereby enhancing opsonophagocytosis, is a promising potential target for therapeutic intervention. PMID- 16242528 TI - C-reactive protein increases matrix metalloproteinase-2 expression and activity in cultured human vascular smooth muscle cells. AB - The C-reactive protein (CRP) is a strong predictor of cardiovascular events both in the general population and in patients with coronary artery disease. We aimed to evaluate whether in cultured human vascular smooth muscle cells (hVSMC) CRP modulates the synthesis and release of metalloproteinase-2 (MMP-2), which is deeply involved in plaque instabilization and vascular remodeling, and of the tissue inhibitor of metalloproteinase-2 (TIMP-2). Both in supernatants and in cell lysates of cultured hVSMC exposed to CRP (0-10 mg/L), we evaluated MMP-2 activity (gelatin zymography), MMP-2 and TIMP-2 protein synthesis (immunoblotting), MMP-2 and TIMP-2 mRNA expression (reverse transcription polymerase chain reaction). CRP effects were also investigated after cell exposure to specific MEK inhibitor PD98059 (15-30 micromol/L) to evaluate the involvement of mitogen-activated protein kinase (MAPK). CRP upregulated MMP-2 mRNA expression. MMP-2 synthesis and activity were increased by 1-10 mg/L CRP starting from 8-hour incubation. The effect was prevented by exposure to PD98059. CRP did not modify TIMP-2 mRNA expression, protein synthesis, and secretion. CRP, at concentrations that predict cardiovascular events, upregulates MMP-2 mRNA expression and increases MMP-2 protein synthesis and release in hVSMC through mechanisms involving activation of MAPK pathway. These data indicate that CRP is not only a risk marker for vascular events, but it is also directly involved in the mechanisms leading to remodeling and instabilization of atherosclerotic plaque. PMID- 16242530 TI - Effects of recombinant-hemoglobin solutions rHb2.0 and rHb1.1 on blood pressure, intestinal blood flow, and gut oxygenation in a rat model of hemorrhagic shock. PMID- 16242529 TI - Tetrathiomolybdate protects against cardiac damage by doxorubicin in mice. AB - Cardiac toxicity is the limiting factor in therapy with doxorubicin, an otherwise useful cancer drug. In this article we detail our study of a mouse model of doxorubicin-induced cardiac toxicity in which, after 4 days' treatment, doxorubicin caused marked increases in plasma concentrations of creatine kinase, lactic dehydrogenase, and troponin I, indicators of cardiac injury; marked increases in the plasma concentrations of tumor necrosis factor-alpha and interleukin-1(beta), both inflammatory cytokines; and a marked increase in the plasma concentration of interleukin-2, an indicator of cytotoxic T-cell activation. Therapy with tetrathiomolybdate, designed to limit copper availability, eliminated almost all of the increases of these six parameters in plasma. The marked protection against cardiac injury by doxorubicin in tetrathiomolybdate-treated animals suggests that tetrathiomolybdate would be of use clinically in helping prevent doxorubicin toxicity in patients. In other preclinical work, it has been shown that tetrathiomolybdate potentiates the chemotherapeutic effect of doxorubicin in cancer, so a double benefit might accrue clinically from the combined use of tetrathiomolybdate and doxorubicin. The mechanism by which tetrathiomolybdate protects against doxorubicin toxicity is of considerable interest. Our working hypothesis, based on the inhibition of interleukin-2 by tetrathiomolybdate as shown here, is that tetrathiomolybdate interrupts the inflammatory cascade at the activated-T-lymphocyte stage. PMID- 16242531 TI - About the cover illustration. PMID- 16242532 TI - Taste responses to dilute sucrose solutions are modulated by stage of the estrous cycle and fenfluramine treatment in female rats. AB - Meal size is decreased during the estrous stage of the rat's ovarian reproductive cycle. This is mediated, in part, by estradiol's ability to increase the strength by which negative-feedback signals function to inhibit meal size. For example, we recently reported that the anorectic effect of fenfluramine, a serotonin agonist, is enhanced during estrus. Here, we investigated whether a decrease in the strength of positive-feedback signals, like those related to the taste of food, contributes to the decrease in meal size observed either in estrous rats or following fenfluramine treatment. Rats were given brief access to six sucrose solutions (0.0, 0.025, 0.05, 0.1, 0.2, and 0.4 M) and the mean number of licks to these solutions was monitored in diestrous and estrous rats treated with 1 mg/kg fenfluramine or saline vehicle. Following saline treatment, estrous rats displayed fewer licks than diestrous rats to the 0.025 M sucrose solution. Following fenfluramine treatment, a decrease in the number of licks to 3 of the 5 sucrose solutions was observed in diestrous rats only. This decrease in sucrose palatability was limited to brief access tests, as overnight preference for the 0.025 M sucrose solution was not decreased by fenfluramine in either diestrous or estrous rats. Our findings suggest that estrous rats experience a decrease in the strength of positive-feedback signals elicited by a dilute sucrose solution and that the anorectic effect of fenfluramine is associated with a decline in positive-feedback signaling in the diestrous rat. PMID- 16242533 TI - Cognitive restructuring and EEG in major depression. AB - Techniques based on cognitive therapy and electroencephalography (EEG) were used to investigate the predictive utility of EEG alpha power with regard to mood improvement. Controls and individuals with major depression participated in four EEG recording blocks. Blocks 1 and 4 were resting baselines. During Block 2, Ss were asked to think about their "most troubling life difficulty." Next, Ss were introduced to cognitive views of depression and techniques used in cognitive therapy. For Block 3, Ss were asked to use these methods to think again about their life difficulty. Ss who reported greater post- than pre-intervention happiness (i.e., "Responders") exhibited greater overall cortical activity than Non-responders. Depressed Responders further exhibited a cortical asymmetry of greater right relative to left activity in frontal areas. The predictive utility of EEG is discussed with regard to identifying individuals who show mood improvement following cognitive restructuring. PMID- 16242534 TI - Impulsivity affects mismatch negativity (MMN) measures of preattentive auditory processing. AB - The current study addresses the relation between impulsive personality and mismatch negativity (MMN) as measure of auditory processing. In a sample of 33 normals, MMN was measured and related to self-reported measures of impulsive behavior. Participants were tested using a passive auditory oddball paradigm while EEG recordings were obtained. It was found that the negative MMN amplitude at the left lateral cluster was negatively correlated with self-reported impulsivity, with high impulsive individuals showing larger MMN amplitudes than low impulsive individuals. This indicates that impulsivity is related to preattentive auditory processing, a process that is not dependent on response execution. In addition to the well-known involvement of top-down control in impulsivity, these findings implicate that the cognitive bottom-up processing of incoming stimuli is associated with individual differences in impulsivity. Since the source of the MMN is thought to be located in the auditory cortex, this suggests involvement of "lower order" temporal lobe processes in impulsive behaviors. PMID- 16242535 TI - Looking at the heart of low and high heart rate variability fearful flyers: self reported anxiety when confronting feared stimuli. AB - Previous research has shown that phobic subjects with low heart rate variability (HRV) are less able to inhibit an inappropriate response when confronted with threatening words compared to phobic subjects with high HRV [Johnsen, B.H., Thayer, J.F., Laberg, J.C., Wormnes, B., Raadal, M., Skaret, E., et al., 2003. Attentional and physiological characteristics of patients with dental anxiety. Journal of Anxiety Disorders, 17, 75-87]. The aim of this study was to evaluate changes in self-reported anxiety when low HRV and high HRV fearful flyers (N=15) and a matched control group (N=15) were exposed to flight-related pictures, flight-related sounds or both pictures and sounds. We hypothesized that sounds would be crucial to evoke fear. Also, low HRV fearful flyers were expected to report higher anxiety than high HRV fearful flyers assuming anxiety as their inappropriate response. Decreases on HRV measures were also predicted for a subgroup of phobic participants (N=10) when confronted with the feared stimuli. Our data supported the hypothesis that sounds are crucial in this kind of phobia. Low HRV fearful flyers reported higher anxiety than high HRV fearful flyers in two out of three aversive conditions. The predicted HRV decreases were not found in this study. Results are discussed in the context of avoidance of exposure based treatments. PMID- 16242536 TI - Blood pressure changes highlight gender differences in emotional reactivity to arousing pictures. AB - The current study was aimed at investigating the effects of gender on the magnitude and patterning of blood pressure responses to specific pleasant and unpleasant, arousing visual stimuli. Systolic and diastolic blood pressure (SBP and DBP), as well as heart rate (HR) and skin conductance (SCR) responses were investigated during picture viewing in 21 female and 25 male students. The pattern of SCR and HR reactivity across emotional categories was found to be similar for men and women. Gender was found to be an effective moderator of BP responses specifically to sexual stimulus content, which prompted greater reactivity in men than in women. These findings extend prior research on gender differences in autonomic responding to emotional visual stimuli and suggest that BP changes might reflect sexual peripheral arousal more than other autonomic measures. PMID- 16242537 TI - Subthreshold amounts of social odorant affect mood, but not behavior, in heterosexual women when tested by a male, but not a female, experimenter. AB - Previously, we have demonstrated that exposure to the endogenous steroid androstadienone has the ability to modulate women's mood in that they feel more focused [Lundstrom, J.N., Goncalves, M., Esteves, F., Olsson, M.J., 2003a. Psychological effects of subthreshold exposure to the putative human pheromone 4,16-androstadien-3-one. Hormones and Behaviour 44 (5), 395-401]. Here, we tested the hypothesis that androstadienone exposure would modulate participating women's mood and corresponding behavior as measured by a sustained attention task. Thirty seven women participated in a double-blind, within-group experiment and were tested by either a female or a male experimenter. Effects on mood, psychophysiological arousal, sustained attention, and ratings of male facial attractiveness were assessed. Sensory detection of the experimental substance was rigorously controlled for by psychophysical testing. The results showed that exposure to a non-detectable amount of androstadienone modulated women's psychophysiological arousal and mood in a positive direction but did not change attention performance or rating of facial attractiveness. Moreover, mood effects were only evident when an experimenter of the opposite sex conducted the testing. This suggests that social context is important for mood effects of androstadienone exposure in women. PMID- 16242539 TI - Repair of thoracoabdominal aortic aneurysms with fenestrated and branched endovascular stent grafts. AB - OBJECTIVE: To report the repair of thoracoabdominal aortic aneurysms (TAAAs) with fenestrated and branched endovascular stent grafts (EVSGs). METHODS: Four patients with asymptomatic TAAAs were treated with custom-designed Zenith fenestrated and branched EVSGs. Three patients had undergone previous open aortic aneurysm repair. Thirteen visceral vessels in four patients were targeted for incorporation by graft fenestrations and branches. RESULTS: The fenestration/orifice interface was secured with balloon-expandable Genesis stents or Jostent stent grafts in 9 of 13 target vessels. Completion angiography demonstrated antegrade perfusion in 12 of 13 target vessels. One renal artery occluded because of graft rotation during deployment. There were no endoleaks. Three patients required additional surgical procedures related to access vessels. One patient required reoperation for bleeding from an extra-anatomic bypass graft and subsequently died from multisystem organ failure. Three patients made an uncomplicated recovery. No patient developed spinal cord ischemia. Computed tomography at 12 months in the 3 survivors demonstrated complete aneurysm exclusion with antegrade perfusion in all 10 target vessels. CONCLUSIONS: TAAA repair with fenestrated and branched EVSGs is feasible and provides an acceptable and promising alternative to conventional surgical repair in selected patients. PMID- 16242540 TI - Emergency endovascular treatment for ruptured abdominal aortic aneurysm and the risk of spinal cord ischemia. AB - BACKGROUND: Spinal cord ischemia is a rare complication after open surgical repair for ruptured abdominal aortic aneurysms (rAAA). The use of emergency endovascular aortic aneurysm repair (eEVAR) is increasing, and paraplegia has been observed in a few patients. The objective of this study was to assess the incidence and pathogenesis of spinal cord ischemia after eEVAR in greater detail. METHODS: This was a retrospective analysis of patients who had eEVAR for rAAA in three hospitals in The Netherlands and Belgium during a 3-year study period that ended in February 2004. The use of aortouniiliac devices combined with a femorofemoral crossover bypass was the preferred technique. Patients with postoperative symptoms of spinal cord ischemia were identified and the influence of potential risk factors was assessed. These factors included the presence of common iliac artery aneurysms necessitating device limb extension to the external iliac artery with associated overlapping the hypogastric artery, the prolonged interruption of bilateral hypogastric artery arterial inflow during the procedure (defined "functional aortic occlusion time" >30 minutes), and the occurrence of preoperative hemodynamic shock. RESULTS: Thirty-five patients were treated by EVAR and they constituted the study group. The first-month mortality in the study group with EVAR was 23%. Four patients (11.5%) with EVAR developed paraplegia postoperatively; the unilateral or bilateral hypogastric artery in all four patients became occluded during the procedure. In the other 31 patients who did not have paraplegia, the unilateral or bilateral hypogastric arteries became occluded in 14 patients (45%). This constituted a significant difference in the prevalence of hypogastric artery occlusion in patients with or without paraplegia (P = .04). The functional aortic occlusion time was prolonged in all four patients with paraplegia and in five without spinal cord ischemia (P = .0003). All four patients with spinal cord ischemia presented with hemodynamic shock. This factor did not reach a significant difference from nonparaplegic patients. CONCLUSION: Emergency EVAR continues to be a promising approach to reduce the high mortality of rAAA, but the incidence of spinal cord ischemia after endovascular treatment of rAAA was worrisome. Although the pathogenesis is most likely multifactorial, interruption of the hypogastric artery inflow appeared to have significant influence. In patients with aneurysmatic common iliac arteries, any effort should be made to minimize hypogastric occlusion time during the procedure and to maintain hypogastric artery inflow afterwards, either by the use of a bell-bottom iliac extension or by electing open repair. PMID- 16242541 TI - Endovascular management of ruptured abdominal aortic aneurysms: 6-year experience from a Greek center. AB - OBJECTIVE: To report our early experience with endovascular treatment of patients with ruptured abdominal aortic aneurysms (RAAAs). METHODS: Between March 1998 and October 2004, 40 consecutive patients with an RAAA presented to our unit; 38 underwent assessment by computed tomography, whereas 2 died on arrival before any assessment and treatment was possible. Twenty-three patients (61%) were suitable for stent grafting, and all proceeded to endovascular repair. Of these, 17 underwent operation with local anesthesia, 1 did so under general anesthesia, and a further 5 procedures were commenced under local anesthesia and converted to general anesthesia. A total of 14 bifurcated and 10 aortouni-iliac stent grafts were implanted; in 1 patient, the bifurcated graft was converted to an aortouni iliac repair during surgery because of technical difficulties. RESULTS: Stent graft deployment in the intended location without a type I or III endoleak was technically successful in 22 of the 23 patients. There were no conversions to open surgery. The 30-day mortality was 39%. Six patients died immediately or soon after the procedure because of severe hypovolemic shock, and three died within 30 days from cardiac causes. After surgery, 13 complications were encountered in 10 patients (3 cardiac, 4 respiratory, 5 renal, and 1 implant related). Two patients required reintervention--one for a type I endoleak and one for limb occlusion. There were 14 survivors. During a median follow-up of 410 days (range, 90-1650 days), 2 more patients died from myocardial infarction, and 9 remain well; 3 patients were lost to follow-up. There were three secondary interventions (two for type I endoleak and one for stent-graft thrombosis). CONCLUSIONS: Endovascular treatment of RAAAs is feasible, and the early experience is promising. More experience and evidence from randomized trials are needed to determine whether such an approach is superior to open surgery. PMID- 16242543 TI - Endovascular repair of abdominal aortic aneurysm in octogenarians: an analysis based on EUROSTAR data. AB - PURPOSE: To investigate the early and late outcome after endovascular treatment of abdominal aortic aneurysm (EVAR) in octogenarians compared with patients aged < 80 years. METHODS: Patients treated for abdominal aortic aneurysm (AAA) with endovascular repair during the period 1996 to 2004 were collated in the EUROSTAR registry. This study group consisted of 697 patients aged > or = 80 years. Comparison was made with 4198 patients aged < 80 years with regard to the incidence of preoperative characteristics and outcomes of the procedure. RESULTS: The proportion of octogenarians treated by EVAR increased during the study period, from 11% in the first year to 18% in the last year. Octogenarians more frequently had cardiac disease, impaired renal function, and pulmonary disease (P = .03, P < .0001 and P = .0001). Thirty-two percent of the octogenarians were recorded unfit for open surgery as opposed to 22% in younger patients (P < .0001); they also had a larger aneurysm diameter (62 vs 58 mm, respectively; P < .0001). The 30-day and in-hospital mortality in octogenarians was 5% vs 2% in the younger group (P < .0001). More device-related complications and systemic complications, including cardiac disease, were noted in octogenarians (7% vs 5% and 19% vs 11%, P = .03 and P < .0001, respectively). This group of patients also had a higher incidence of postoperative hemorrhagic complications, including hematoma (7% vs 3%, P < .0001, respectively). No differences in conversion to open repair and post-EVAR rupture rate were observed. Aneurysm-related mortality and late all-cause mortality was 7% vs 3% and 10% vs 7%, both P < .0001. CONCLUSION: Our study supports that EVAR might be considered when treating elderly patients, provided their aneurysms are anatomically suited for the endovascular technique. The risk for late complications compared with open repair may be outweighed by a lower early mortality as well as a shorter time for physical recovery. PMID- 16242545 TI - Secondary conversion of the Gore Excluder to operative abdominal aortic aneurysm repair. AB - OBJECTIVE: Reports continue to document the occurrence of major adverse events after endovascular aortic aneurysm repair. Although many of these problems can be successfully managed through endovascular salvage, operative conversion with explantation of the endoprosthesis remains necessary in some patients. We report herein a review of all patients initially enrolled in multicenter US clinical trials of the Excluder endograft who underwent secondary conversion to open surgical repair. METHODS: Clinical data and relevant medical records of patients enrolled in phase I and II multicenter US clinical trials of the Excluder endograft were retrospectively reviewed for adverse events and further narrowed to those patients who underwent secondary operative conversion. Hospital records, operative and anesthesia reports, and all imaging studies were analyzed at initial implantation and at the time of subsequent open surgical repair. RESULTS: Late open conversion was performed in 16 (2.7%) of the 594 patients enrolled in the Excluder clinical trials. Presumed endotension accounted for 8 of 16 of secondary conversions. In two of these patients, however, an endoleak was identified at the time of open surgical repair. Of the remaining eight patients, two underwent conversion for device infection, five for persistent endoleak, and one for aneurysm rupture. The overall 30-day mortality was 6.25% (1/16), with one death occurring in a patient with a ruptured aneurysm. Of patients who underwent conversion because of endotension, the maximal abdominal aortic aneurysm diameter (mean +/- SD) at the time of initial implantation and subsequent graft removal was 61 +/- 11 mm and 70 +/- 10 mm, respectively. The mean time to open conversion for treatment of endotension was 37 +/- 12 months (range, 20-50 months; median, 42 months). Freedom from conversion was 98.6% and 96.7% at 24 and 48 months, respectively. CONCLUSIONS: Endotension in the absence of a demonstrable endoleak has been a major indication for late surgical conversion in patients treated with the Excluder endograft. Given the potential presence of an undetected endoleak and the possible effects of progressive sac enlargement on long-term device stability, continued close surveillance of patients with assumed endotension is required. Should changes in device design eliminate endotension, a further reduction in the already low incidence of late open conversion of the Excluder endograft can be anticipated. PMID- 16242546 TI - Absence of proximal neck dilatation and graft migration after endovascular aneurysm repair with balloon-expandable stent-based endografts. AB - OBJECTIVE: Proximal neck dilatation (PND) and/or endograft migration with the subsequent development of type I endoleak is a significant cause of late endograft failure after endovascular abdominal aortic aneurysm repair (EVAR). Although there are numerous reports examining PND in patients receiving endografts that use self-expanding stents (SES) for proximal fixation, there are no such reports for patients treated with endografts that use balloon-expanding stents (BES). The purpose of this study was to investigate PND and endograft migration after EVAR with BES endografts. METHODS: We retrospectively reviewed all charts and all serial computed tomographic scans available for patients who underwent EVAR with a BES endograft (surgeon-made, aortounifemoral polytetrafluoroethylene graft with a proximal Palmaz stent) between August 1997 and October 2002. Only patients with longer than a 12-month follow-up were analyzed. Neck diameter was measured at the level of the lowest renal artery and at 5 mm below it. PND was defined as neck enlargement of 2.5 mm or more. To assess endograft migration, the distance between the superior mesenteric artery and the cranial end of the BES was measured. Stent migration was defined as a change of 5 mm or more. RESULTS: A total of 77 patients received this device during the study period. The technical success rate was 99%. The 1-, 3-, and 5 year survival was 66%, 48%, and 29.5%, respectively. Complete serial computed tomographic scans were available in 41 of the 48 patients who survived 12 months or longer after the operation. The mean follow-up period for these patients was 31 months (range, 12-66 months). The maximum aneurysm diameter was either unchanged or decreased in 35 patients (85%). The immediate postoperative proximal neck diameter was 19 to 29 mm (median, 24 mm). This was unchanged at the latest follow-up. None of the patients had significant PND. The cranial end of the BES was located in the area between 14 mm proximal and 36 mm distal to the superior mesenteric artery (median, 6 mm). None of the patients developed significant endograft migration. CONCLUSIONS: Neither PND nor endograft migration was observed with the BES endograft. The nature of the SES may be responsible for the observed neck dilatation and device migration after EVAR with SES endografts. This study suggests that BES may be a better fixation method for EVAR. PMID- 16242547 TI - Endovascular treatment for aortic disease: is a surgical environment necessary? AB - OBJECTIVE: Interventional radiologists, cardiologists, and vascular surgeons are capable of performing endovascular procedures successfully in their respective environments. Suboptimal anatomy or intraoperative technical problems can be encountered, and endovascular management alone is not always suitable. The objectives of this study were to define the incidence of adjunctive surgical techniques, to discuss the rationale for endovascular reconstruction in a well developed surgical environment, and to assess the effect of experience on the incidence of adjunctive repair. METHODS: All primary aortic and aortoiliac elective, urgent, and emergent endovascular procedures performed at the Peter Lougheed Center and entered into a prospective database from May 25, 1999 to June 01, 2005, were reviewed. All adjunctive surgical techniques to enable stent deployment, enhance attachment site, or solve intraoperative difficulties were captured. The study period was divided into two time periods based on learning curve data to assess the effect of experience on the rate of adjunctive repairs. RESULTS: Four hundred thirty-eight patients underwent elective (80%), urgent (15%), or emergent (5%) endovascular procedures during the study period. These consisted of 101 thoracic and 337 abdominal operations, including the use of 13 fenestrated stents. One hundred thirty-nine patients (31.7%) required 180 open surgical procedures. Complete data were available for the entire patient cohort. The mean follow-up was 793.2 days (SD, 519.1 days). Procedures were necessary for vascular access, arterial dissection/rupture, limb ischemia, and enhancement/elongation of the stent attachment site. The persistent endoleak rate was 5.3%, the late rupture rate was 0.7%, the conversion rate was 1.6%, the 30 day surgical mortality rate was 3.2%, all-cause mortality to date is 7.3%, and the reintervention rate was 4.6%. There was no statistically significant effect of the learning curve on the incidence of surgical adjunctive procedures in either the thoracic group (11/26 [42.3%] for phase 1 vs 17/75 [22.6%] for phase 2) or the abdominal group (14/50 [28.0%] for phase 1 vs 97/287 [33.8%] for phase 2). Overall, 31.5% of patients required adjunctive surgical repair. CONCLUSIONS: Successful endografting requires endovascular expertise in addition to a well developed surgical environment to increase applicability and decrease patient risk. Despite advances in endovascular technology, hybrid techniques will continue to be required to achieve good overall success rates. PMID- 16242549 TI - Is abdominal aortic aneurysm repair appropriate in oxygen-dependent chronic obstructive pulmonary disease patients? AB - BACKGROUND: The life expectancy of patients with oxygen-dependent chronic obstructive pulmonary disease (COPD) is significantly reduced, but the risk of any intervention is considered prohibitive. However, severe COPD may increase the risk of abdominal aortic aneurysm (AAA) rupture. We reviewed our experience with AAA repair in oxygen-dependent patients to determine whether operative risk and expected long-term survival justify surgical intervention. METHODS: A retrospective review of 44 consecutive patients with oxygen-dependent COPD undergoing AAA repair over an 8-year period was performed. Information was recorded for survival, length of follow-up, patient age, medical comorbidities, pulmonary function tests, and operative approach. Survival data were analyzed by Kaplan-Meier curves and compared with published cohorts of oxygen-dependent patients and the natural history of untreated aneurysms. RESULTS: Twenty-four patients underwent endovascular aneurysm repair (EVAR), and 20 underwent open procedures (14 retroperitoneal and 6 transabdominal). The mean AAA diameter was 6.1 cm (range, 5-9.5 cm). The mean age was 71.4 years, and 82% of patients were male. Operative mortality was 0%. The mean length of stay was 11.2 days for open procedures and 4.3 days for EVAR (significantly longer than that for standard risk patients). The mean survival time was 37.9 months (range, 2-91 months). Preoperative medical comorbidities, type of repair, and pulmonary function tests were not predictive of survival. Postoperative morbidity was significantly higher with open repair. Long term survival was comparable to historical series of the natural history of O2 dependent patients without AAA but better than untreated 6 cm AAA cohorts. At 42 months, almost 50% of patients in our study group were still alive, compared to 20% survival at 34 months for those with untreated 6 cm AAAs. CONCLUSIONS: It is reasonable to continue to offer AAA repair to home oxygen-dependent COPD patients who are ambulatory and medically optimized and who are without untreated coronary artery disease. Although EVAR may be the most suitable treatment for oxygen-dependent COPD patients, our results show that even open repair may be safely performed in this population, with acceptable results. PMID- 16242550 TI - Evidence for early nasogastric tube removal after infrarenal aortic surgery: a randomized trial. AB - OBJECTIVE: Nasogastric tube (NGT) decompression after abdominal surgery is still largely used to prevent nausea and vomiting. However, indications are based more on practice than on studies. Moreover, prolonged NGT decompression can lead to complications. In this prospective and randomized study, we evaluated the effects of early withdrawal of NGTs in patients undergoing surgery of the infrarenal aorta. METHODS: Between October 2001 and May 2002, consecutive patients underwent scheduled infrarenal aortic operations. Patients were prospectively randomised into two groups: group 1, NGT maintenance until the passage of flatus; and group 2, NGT removal at the time of tracheal extubation. Preoperative and perioperative data were collected. The main end point was the occurrence of nausea and vomiting. Secondary end points were tolerance of NGT withdrawal and postoperative complications. Criteria were compared between groups by using Mann-Whitney or Fisher exact tests. RESULTS: Forty-six patients underwent aortic operations. Six patients were subsequently excluded from the study. Of the 40 randomized subjects, 20 patients were included in each group. Preoperative and intraoperative data were similar in both groups. There was no statistical difference between groups regarding nausea and vomiting. In group 1, the occurrence of respiratory complications was more frequent compared with group 2 (5 vs 0 complications; P = .023). There was no significant difference in intensive care unit stay, but the hospital stay was shorter in group 2 (mean, 9 +/- 3 days vs 15 +/- 9 days; P = .016). There were no differences in other adverse events. CONCLUSIONS: This study does not support a significant effect of early removal of NGTs on nausea and vomiting in patients undergoing open repair of the infrarenal aorta. However, these findings suggest that NGT maintenance increases the risk of respiratory complications and the length of hospital stay. PMID- 16242551 TI - Improved outcomes in the recent management of secondary aortoenteric fistula. AB - OBJECTIVE: We reviewed the presentation, management, and late events in a recent experience with aortic graft-enteric communications (secondary aortoenteric fistula) to identify variables associated with poor outcomes. METHODS: Since 1991, 29 patients (19 men, 10 women; mean age, 70) presented with a secondary aortoenteric fistula. The duration from aortic graft implantation to aortoenteric fistula development ranged from 8 months to 180 months (mean, 47 months). Presenting symptoms included gastrointestinal bleeding (n = 25), sepsis and retroperitoneal abscess (n = 3), and ruptured para-anastomotic aneurysm (n = 1). One third (10/29) of patients were hypotensive at presentation. Repair was accomplished by graft excision with axillobifemoral bypass (17 simultaneous, 8 staged), graft excision with in situ deep vein replacement (n = 2), or graft excision with rifampin-treated prosthetic replacement (n = 2). Mean follow-up was 51 months, and the incidence of late events was reported by life table methods. RESULTS: The operative (< or = 30-day) mortality rate was 21%, with shock at presentation (P < .01), the need for preoperative transfusion (P < .01), and use of suprarenal aortic clamping during aortoenteric fistula repair (P = .03) associated with lethal outcome. Cumulative mortality related to aortoenteric fistula management was only 24% within an overall 5-year survival rate of 61%. Freedom from recurrent infection or amputation was 86% and 88% at 5 years, respectively, and assisted graft patency was 79%. Presence of staphylococcal species in 22% of patients and occasional operative findings of adjacent perigraft "biofilm" suggests that underlying indolent graft infection may contribute to the development of aortoenteric fistula in some cases. CONCLUSION: Outcomes associated with aortoenteric fistula repair were surprisingly favorable in the absence of preoperative hemodynamic instability. PMID- 16242552 TI - Graft-associated hemorrhage from femoropopliteal vein grafts. AB - OBJECTIVE: The femoropopliteal vein (FPV) graft has been used extensively for large-caliber vascular reconstructions. To date, there have been no reports of anastomotic dehiscence or rupture leading to graft-associated hemorrhage (GAH). In the present report, we review our experience with GAH from FPV grafts to determine the incidence of this problem, to better understand the etiology, and to determine potential methods to prevent this complication. METHODS: All patients undergoing arterial reconstructions with FPV grafts were entered into a registry that included demographics, operative details, complications, and follow up information. Episodes of GAH that occurred during the period from 1990 to 2004 were studied to determine etiologic factors and outcomes. RESULTS: During the study period, 574 FPV grafts were used for arterial reconstructions in 364 patients. GAH occurred in 11 patients (3%). Onset of GAH ranged from 1 hour to 180 days after operation. The mean blood transfusion requirement for GAH was 10 +/- 4 units. In three patients, the etiology of GAH was purely technical, resulting in a slipped or "popped" tie from a large side branch. In eight patients, the etiology was due to graft disruption secondary to uncontrolled infection and failure of anastomotic healing. Most of these patients were being treated for aortic graft infection. Special risk factors for this complication included malnutrition, ongoing polymicrobial and fungal infections, immunocompromised state, active cancer, steroid treatment, and ongoing graft contamination from gastrointestinal or pharyngeal leaks. Outcomes included four deaths and one stroke. CONCLUSIONS: GAH is a serious complication with high morbidity, mortality, and transfusion requirements. Although technical problems are preventable, FPV grafts, like all biologic grafts, can develop disruption with GAH from ongoing infection, especially in severely immunocompromised patients who are malnourished and have poor healing ability. Strategies for prevention and alternative treatment modalities are appropriate in patients at high risk for GAH. PMID- 16242553 TI - The surgical treatment of arterial aneurysms in Behcet disease: a report of 16 patients. AB - OBJECTIVE: The aim of this article is to report our experience in the surgical treatment of arterial aneurysms in patients with Behcet disease. METHODS: From October 2001 through May 2004, 18 arterial aneurysms were diagnosed in 16 Behcet patients. All patients were male. The patients ranged in age from 24 to 52 years (mean, 37.4 +/- 5.2 years). There were six abdominal aortic, three common femoral, two iliac, two popliteal, two superficial femoral, and two anterior tibial aneurysms and one subclavian artery aneurysm. All patients but four were in remission at the time of diagnosis. Those 4 patients received immunosuppressive therapy before the surgical intervention to induce remission. After hospital discharge, all patients were followed up regularly at 3-month intervals. The mean duration of follow-up was 17 +/- 4.2 months (range, 6-24 months). RESULTS: All patients underwent a successful surgical intervention. During the study period, we performed five aortic tube graft interpositions, two aortofemoral bypasses, one aortobifemoral bypass, three common femoral artery graft interpositions, and two femoropopliteal bypasses. The popliteal artery (n = 2), anterior tibial artery (n = 2), and subclavian artery (n = 1) aneurysms were repaired primarily. There was no in-hospital mortality. One patient with an abdominal aortic aneurysm had to undergo reoperation because of postoperative bleeding. The postoperative hospital stay was 8.5 +/- 4.3 days. Two patients were lost to follow-up. During the follow-up period, two false aneurysms of the common femoral artery were repaired with a graft interposition procedure. Another patient who had undergone an aortic tube graft interposition was readmitted 9 months later with an external iliac artery aneurysm. The external iliac artery was ligated through a retroperitoneal approach, and femorofemoral bypass was performed. In addition, one femoropopliteal interposition graft was occluded, without disabling ischemia. CONCLUSIONS: Although aneurysmal disease is rare in Behcet disease, it can complicate the clinical picture and cause life-threatening complications. We believe that the establishment of remission before the surgical intervention decreases the incidence of postoperative complications. Because recurrence at the site of anastomosis is possible, prolonged monitoring is required. PMID- 16242554 TI - Role of computed tomographic angiography in the detection and comprehensive evaluation of persistent sciatic artery. AB - PURPOSE: To define the role of computed tomographic (CT) angiography in the evaluation of persistent sciatic artery and to identify its potential advantages as a diagnostic modality. METHODS: Between July 2002 and August 2004, 307 consecutive patients underwent CT angiography for suspected lower-extremity arterial insufficiency. All CT angiograms were retrospectively reviewed to determine the presence and laterality of persistent sciatic artery and its associated vascular abnormalities, such as aneurysm, thrombus, distal thromboembolism, and atherosclerotic change. The relationship of persistent sciatic artery with adjacent structures, such as sciatic nerve, muscle, accompanying vein, and femoral artery, as well as the presence of other anomalies, was analyzed. Clinical data regarding the presenting symptoms and hospital course were obtained from patient charts. RESULTS: Six persistent sciatic arteries, with or without occlusion, were identified in five female patients (age range, 54 to 80 years). CT angiography revealed unilateral persistent sciatic artery in four patients (left, 3; right, 1) and bilateral persistent sciatic artery in one patient. Aneurysm was present in two (mean size, 26 mm x 20 mm), thrombosis in three, and distal thromboembolism in all six persistent sciatic arteries. All persistent sciatic arteries coursed along the sciatic nerve and continued as popliteal artery. Characteristically, in all these instances, the superficial femoral arteries were hypoplastic and tapered smoothly. Anomalous popliteal venous drainage was noted in all ipsilateral limbs with persistent sciatic artery and even in contralateral limbs with normal superficial femoral artery in all but one. CONCLUSION: CT angiography enables the detection of persistent sciatic artery even in the presence of complete occlusion and is useful in the comprehensive evaluation of various complications and associated venous anomalies. It can potentially be used as the sole imaging modality for persistent sciatic artery. PMID- 16242555 TI - Correlation of carotid artery stump pressure and neurologic changes during 474 carotid endarterectomies performed in awake patients. AB - PURPOSE: A carotid artery stump pressure (SP) of < 50 mm Hg and abnormal electroencephalography (EEG) changes have been suggested as indications for selective shunting in patients undergoing carotid endarterectomy (CEA) under general anesthesia. We attempted to determine the optimal SP threshold that correlated with neurologic changes in awake patients undergoing CEA using cervical block anesthesia (CBA) and performed a cost comparison with EEG monitoring. METHODS: Between July 1, 1995, and December 31, 2004, SP was measured during 474 CEAs performed under CBA by inserting a 19-gauge butterfly needle into the common carotid artery. A saline-filled intravenous bag in the patient's contralateral hand was connected to pressure tubing to generate waveforms with hand squeezing that could be visualized on a monitor. Systemic pressure was maintained approximately 10 mm Hg higher than baseline. Accurate SPs were confirmed by the finding of flatline waveforms after internal carotid artery clamping. Selective shunting was performed when neurologic changes occurred (aphasia, inability to squeeze the contralateral hand, decreased consciousness), regardless of SP. During this same period, 142 patients underwent CEA using GA, and SP was also measured. RESULTS: Shunting was necessary because of neurologic changes in 7.2% (34/474) of all CEAs performed using CBA: 0.9% (3/335) with SPs > or = 50 mm Hg systolic vs 1.0% (4/402) with SPs > or = 40 mm Hg systolic, and 22% (31/139) with SPs < 50 mm Hg systolic vs 42% (30/72) with SPs < 40 mm Hg systolic. If these 474 CEAs had been performed using GA, shunts would have been used in 29% (139/474) of patients for a SP < 50 mm Hg systolic vs 15% (72/474) for a SP < 40 mm Hg systolic. In patients not shunted, the perioperative stroke/death rate was 1.2% in patients (4/332) with SPs > or = 50 mm Hg vs 1.0% (4/398) with SPs > or = 40 mm Hg. Three of the four strokes occurred > 24 hours postoperatively and were unrelated to lack of shunting and ischemia. There was no significant difference in the percentage of patients with SPs > or = 50 mm Hg who underwent CEA using CBA (70%, 335/474) vs GA (67%, 96/142) during this time period. At our hospital, charges for SPe measurement, including anesthesia charges and tubing, were 229 dollars per case vs 3439 dollars per case for EEG monitoring. Use of SP measurements in these 474 patients would have resulted in reduced charges of 1,521,540 dollars compared with EEG monitoring if CEA had been performed under GA. CONCLUSION: Using 40 mm Hg systolic as a threshold, the need for shunting (15%) and the false-negative rate (1.0%) for SP in our series were equivalent to the results of EEG monitoring during CEA reported in the literature. However, charges for SP measurements are dramatically lower compared with EEG monitoring. Our results suggest that a carotid artery SP > or = 40 mm Hg systolic may be considered as an equally reliable but more cost-effective method to predict the need for carotid shunting during CEA under GA compared with EEG monitoring, but further investigation is warranted. PMID- 16242556 TI - Controlled reperfusion using a simplified perfusion system preserves function after acute and persistent limb ischemia: a preliminary study. AB - OBJECTIVE: Reperfusion of the limb after acute and persistent ischemia is associated with high rates of morbidity and mortality despite complete revascularization. Although reperfusion is a prerequisite for maintaining limb function, it may in itself cause further injury. There is experimental evidence that modification of the initial reperfusion modalities can minimize this reperfusion injury. We hypothesized that controlled reperfusion using a simple blood bag perfusion system reduces reperfusion injury and facilitates the return of normal function. METHODS: Fifteen consecutive patients (mean age, 80.5 +/- 5.0 years) with severe, acute lower-limb ischemia were allocated to two treatment arms in this prospective, controlled observational study. Group I (n = 8) underwent surgical embolectomy alone, and group II (n = 7) underwent surgical embolectomy plus controlled reperfusion using a simplified perfusion system. Indication for controlled reperfusion was made by the responsible surgeon. Controlled reperfusion consisted of a 30-minute infusion of a crystalloid reperfusion solution that was mixed with oxygenated blood (the blood:reperfusion solution ratio was 6:1) distal to the occlusion. Duration of ischemia, postoperative amputation rate, motor function of the ischemic limb, and pre- and postoperative serum creatine kinase levels were assessed. RESULTS: The duration of ischemia was 10.7 +/- 1.1 hours in group I and 19 +/- 5.2 hours in group II (P < .05). The site of the arterial occlusion was the iliac artery in nine patients and the common femoral artery in six patients. Full recovery was achieved in six of seven patients in group II and in only two of eight patients in group I (P < .05). There were three in-hospital deaths in group I, and two patients underwent major amputations. No in-hospital deaths or major amputations occurred in group II. CONCLUSION: The results from this preliminary study strongly suggest the hypothesis that the results of conventional embolectomy for acute, severe lower limb ischemia can be improved by controlled reperfusion. To prove our preliminary findings, a large randomized, prospective, controlled, multicenter trial, the Controlled Reperfusion of the Acutely Ischemic Limb trial (CRAIL-Trial) is currently being conducted to prove our preliminary findings. PMID- 16242557 TI - Comparative analysis of autogenous infrainguinal bypass grafts in African Americans and Caucasians: the association of race with graft function and limb salvage. AB - OBJECTIVE: African Americans (AAs) are at risk for developing diabetes mellitus and atherosclerosis. Whether race influences the results of infrainguinal arterial reconstruction is unclear. The purpose of this study was to compare the results of autogenous infrainguinal bypasses in AAs and Caucasians to determine the association of race with graft function and limb salvage. METHODS: This was a retrospective, comparative cohort study of AA and Caucasian patients who had undergone autogenous infrainguinal bypass surgery. Only single-limb bypasses in each patient cohort were considered in this analysis. In patients who had undergone bilateral lower limb bypasses, the first limb bypass was chosen as the index bypass procedure. RESULTS: From January 1985 to December 2003, 1459 autogenous infrainguinal bypasses were performed in 1459 patients for lower limb ischemia. Within this group, 89 AA patients/vein grafts formed the study cohort. The control group comprised 1370 Caucasian patients/vein grafts. Compared with the Caucasian cohort, AA patients were significantly younger (median age, 65 vs 70 years, respectively; P = .001) and predominantly female (57% vs 41%, respectively; P = .002). AA patients also had a higher prevalence of diabetes mellitus, hypertension, cerebrovascular disease, congestive heart failure, and dialysis-dependent renal failure. More AA than Caucasian patients presented with gangrene (34% vs 16%, respectively; P = .001), and more underwent bypass surgery for limb salvage indications (91% vs 81%, respectively; P = .01). The venous conduit used was predominantly the greater saphenous vein (AA, 83%; Caucasian, 85%), and the site of distal anastomosis was at the tibial/pedal level in 67% of AA and 61% of Caucasian patients. Overall morbidity (AA, 28%; Caucasian, 23%) and 30-day mortality (AA, 3%; Caucasian, 3%) were similar. Thirty-day graft failure was significantly greater in AAs than Caucasians (12% vs 5%, respectively; P = .003). The overall 5-year primary graft patency (+/-SE) was significantly worse in AA patients (AA, 52% +/- 6%; Caucasian, 67% +/- 2%; P = .009). The 5-year limb salvage rate (+/-SE) was also significantly worse in AA patients (AA, 81% +/- 5%; Caucasian, 90% +/- 1%; P = .04). With the Cox proportional hazard model, significant risk factors associated with primary graft failure were AA race, age younger than 65 years, female sex, secondary reconstructions, tibial bypasses, and critical limb ischemia. Significant risk factors associated with limb loss were age younger than 65 years, female sex, absence of coronary disease, presence of critical limb ischemia, and secondary reconstructions. CONCLUSIONS: Autogenous infrainguinal bypass surgery in AAs is associated with poorer primary graft patency and limb salvage rates compared with those of Caucasians. This may partially account for the higher rate of limb loss in AA patients with peripheral arterial occlusive disease. PMID- 16242558 TI - The effect of exercise intensity on the response to exercise rehabilitation in patients with intermittent claudication. AB - PURPOSE: The purpose of this randomized trial was to compare the efficacy of a low-intensity exercise rehabilitation program vs a high-intensity program in changing physical function, peripheral circulation, and health-related quality of life in peripheral arterial disease (PAD) patients limited by intermittent claudication. METHODS: Thirty-one patients randomized to low-intensity exercise rehabilitation and 33 patients randomized to high-intensity exercise rehabilitation completed the study. The 6-month exercise rehabilitation programs consisted of intermittent treadmill walking to near maximal claudication pain 3 days per week at either 40% (low-intensity group) or 80% (high-intensity group) of maximal exercise capacity. Total work performed in the two training regimens was similar by having the patients in the low-intensity group exercise for a longer duration than patients in the high-intensity group. Measurements of physical function, peripheral circulation, and health-related quality of life were obtained on each patient before and after the rehabilitation programs. RESULTS: After the exercise rehabilitation programs, patients in the two groups had similar improvements in these measures. Initial claudication distance increased by 109% in the low-intensity group (P < .01) and by 109% in the high intensity group (P < .01), and absolute claudication distance increased by 61% (P < 0.01) and 63% (P < .01) in the low-intensity and high-intensity groups, respectively. Furthermore, both exercise programs elicited improvements (P < .05) in peak oxygen uptake, ischemic window, and health-related quality of life. CONCLUSION: The efficacy of low-intensity exercise rehabilitation is similar to high-intensity rehabilitation in improving markers of functional independence in PAD patients limited by intermittent claudication, provided that a few additional minutes of walking is accomplished to elicit a similar volume of exercise. PMID- 16242559 TI - Hemodynamic effects of intermittent pneumatic compression in patients with critical limb ischemia. AB - BACKGROUND: Traditional teaching assumes that the distal arterial tree is maximally dilated in patients with critical limb ischemia (CLI). Endovascular or arterial bypass procedures are the commonly used interventions to increase distal perfusion. However, other forms of treatment such as spinal cord stimulation or intermittent pneumatic compression (IPC) have been shown to improve limb salvage rates. This prospective study was designed to determine if the use of IPC increases popliteal, gastrocnemial, collateral arterial, and skin blood flow in patients with CLI. METHODS: Twenty limbs with CLI in 20 patients (mean age, 74 years) were evaluated with duplex ultrasound scans and laser Doppler fluxmetry in the semi-erect position before, during, and after IPC. One pneumatic cuff was applied on the foot and the other on the calf. The maximum inflation pressure was 120 mm Hg and was applied for 3 seconds at three cycles per minute. All patients had at least two-level disease by arteriography. Fourteen limbs were characterized as inoperable, and six were considered marginal for reconstruction. Flow volumes were measured in the popliteal, medial gastrocnemial, and a genicular collateral artery. Skin blood flux was measured on the dorsum of the foot at the same time. RESULTS: Significant flow increase during the application of IPC was found in all three arteries (18/20 limbs) compared with baseline values (P < .02). The highest change was seen in the popliteal, followed by the gastrocnemial and the collateral artery. After the cessation of IPC, the flow returned to baseline. This was attributed to the elevation of time average velocity, as the diameter of the arteries remained unchanged. The skin blood flux increased significantly as well (P < .03). In the two limbs without an increase in the arterial or skin blood flow, significant popliteal vein reflux was found. Both limbs were amputated shortly after. CONCLUSIONS: IPC increases axial, muscular, collateral, and skin blood flow in patients with CLI and may be beneficial to those who are not candidates for revascularization. Patients with significant venous reflux may not benefit from IPC. This supports the theory that one of the mechanisms by which IPC enhances flow is by increasing the arteriovenous pressure gradient. PMID- 16242560 TI - Duration and amplitude decay of acute arterial leg inflow enhancement with intermittent pneumatic leg compression: an insight into the implicated physiologic mechanisms. AB - PURPOSE: By acutely enhancing the arterial leg inflow, intermittent pneumatic leg compression (IPC) improves the walking ability, arterial hemodynamics, and quality of life of claudicants. We quantified the duration of acute leg inflow enhancement with IPC of the foot (IPC(foot)), calf (IPC(calf)), or both (IPC(foot+calf)) and its amplitude decay in claudicants and controls in relation to the pulsatility index, an estimate of peripheral resistance. These findings are cross-correlated with the features of the three implicated physiologic mechanisms: (1) an increase in the arteriovenous pressure gradient, (2) suspension of peripheral sympathetic autoregulation, and (3) enhanced release of nitric oxide with flow and shear-stress increase. METHODS: Twenty-six limbs of 24 claudicants with superficial femoral artery occlusion or stenoses (>75%) and 24 limbs of 20 healthy controls matched for age and sex, meeting stringent selection criteria, had their popliteal volume flow and pulsating index (peak-to-peak velocity/mean velocity) measured with duplex scanning at rest and upon delivery of IPC. Spectral waveforms were analyzed for 50 seconds after IPC delivery per 5 second segments. The three IPC modes were applied in a true crossover design. Data analysis was performed with the Page, Friedman, Wilcoxon, Mann-Whitney and chi2 tests. RESULTS: The median duration of flow enhancement in claudicants exceeded 50 seconds with IPC(foot), IPC(calf), and IPC(foot+calf) but was shorter (P < .001) in the controls (32.5 to 40 seconds). Among the three IPC modes, the duration of flow enhancement differed (P < .05) only between IPC(foot) and IPC(foot+calf). After reaching its peak within 5 seconds of IPC, flow enhancement decayed at rates decreasing over time (trend, P < .05, Page test), which in both groups were highest at 5 to 20 seconds, moderate at 20 to 35 seconds, and lowest at 35 to 50 seconds (P < .05, Friedman test). Baseline and peak flow with all IPC modes was similar between the two groups. Pulsatility index attenuation in claudicating limbs lasted a median 32.5 seconds with IPC(foot), 37.5 seconds with IPC(calf), and 40 seconds with IPC(foot+calf); duration of pulsatility index attenuation was shorter in the control limbs with IPC(foot) (30 seconds), IPC(calf) (32.5 seconds), or IPC(foot+calf) (35 seconds), yet differences, as well as those among the 3 IPC modes, were not significant. CONCLUSION: Leg inflow enhancement with IPC exceeds 50 seconds in claudicants and lasts 32.5 to 40 seconds in the controls. Peak flow occurs concurrently with maximal pulsatility index attenuation, within 5 seconds of IPC. Irrespective of group or IPC mode, the decay rate (%) of flow enhancement is highest within 5 to 20 seconds of IPC, moderate at 20 to 35 seconds, and lowest at 35 to 50 seconds. Since attenuation in peripheral resistance terminates with the mid time period (20 to 35 seconds) of flow decay, and nitric oxide has a half-life of <7 to 10 seconds, the study's data indicate that all implicated physiologic mechanisms (1, 2, and 3) are likely active immediately after IPC delivery (0 to 20 sec) and all but nitric oxide are effective in the mid time period (20 to 35 seconds). As the pulsatility index has returned to baseline, the late phase of flow enhancement (35 to 50 seconds) could be attributable to the declining arteriovenous pressure gradient alone. PMID- 16242561 TI - Treatment of venous thromboembolism: adherence to guidelines and impact of physician knowledge, attitudes, and beliefs. AB - OBJECTIVES: To assess the treatment of venous thromboembolism (VTE) in hospitalized patients enrolled in a national, multicenter database. METHODS: This was a retrospective, cohort study that randomly selected VTE patients from 38 academic/teaching, community, and Veterans Administration (VA) hospitals. The study included a physician survey component. The patients selected were those treated between January 2002 and June 2003 who had an ICD-9-CM code for pulmonary embolus (PE), deep vein thrombosis (DVT), or pregnancy-related PE or DVT. RESULTS: The study included 939 patients: 52.7% with DVT, 28.4% with PE, and 18.8% with PE and DVT. Mean age was 59.5 years. Risk factors included obesity (body mass index > 30) in 30.1%, history of VTE in 28.0%, malignancy in 27.4%, surgery in 21.1%, and immobility in 18.5%. Only 56.1% of patients were treated with low-molecular-weight heparin (LMWH). Bridging from LMWH or unfractionated heparin (UFH) to warfarin was completed during hospitalization in 486 (68.6%), but only 246 (50.6%) had an international normalized ratio (INR) > or = 2 for 48 hours before discontinuation of the injectable anticoagulant. Length of stay in patients discharged on bridge therapy was 4.0 +/- 3.7 days vs 8.1 +/- 5.8 days for patients discharged on warfarin therapy (P < .001). Ninety-two (10.1%) patients were discharged with neither oral nor injectable anticoagulation and had a mean duration of treatment of only 10.6 +/- 16.2 days. Of 245 physicians surveyed from participating hospitals, 84% and 53%, respectively, indicated that LMWH was their preferred agent for treatment of DVT and treatment of PE. With regard to warfarin, 30% did not believe it was necessary to have a therapeutic INR for > or = 2 days before discontinuing LMWH or UFH, and 27% responded that it was necessary to keep DVT patients in the hospital until they were therapeutic. CONCLUSIONS: In this cross-section of United States hospitals, lower than anticipated use of LMWH, insufficient bridging from UFH or LMWH to warfarin, and continuation of anticoagulation after hospitalization were all problems discovered with the treatment of VTE. Physician knowledge, attitudes, and beliefs are partially responsible for the gap between actual practice and international guidelines. These results suggest that hospitals should evaluate their adherence to international VTE treatment guidelines and develop strategies to optimize antithrombotic therapy. PMID- 16242562 TI - Calf compression pressure required to achieve venous closure from supine to standing positions. AB - BACKGROUND: Compression therapy needs to narrow the veins of patients with venous disorders to achieve a hemodynamic effect. This study investigated the external pressure necessary to narrow and occlude leg veins in different body positions. METHODS: In nine healthy volunteers and five patients with incompetent small saphenous veins, the diameter of the small saphenous veins and of one posterior tibial vein was measured at the mid level of the calf by duplex ultrasound scans in sitting, standing, and supine positions. A modified blood pressure cuff with an acetate window that permitted ultrasound visualization of the veins was gradually inflated, and the pressures needed to narrow or occlude the veins were recorded. RESULTS: Initial narrowing occurs with a median pressure of between 30 and 40 mm Hg in the sitting and standing positions. Complete occlusion of superficial and deep leg veins occurs with 20 to 25 mm Hg in the supine position, between 50 and 60 mm Hg in the sitting position, and at about 70 mm Hg in the standing position. The difference between sitting and standing, and between standing and supine, was statistically significant (P < .01 and P < .001, respectively). CONCLUSION: The external pressure has to exceed the hydrostatic pressure in the vein to compress leg veins effectively. Higher external pressures than can be expected to be delivered by elastic compression stockings are required to achieve the collapse of lower-extremity veins in the upright position. PMID- 16242563 TI - Stenosis detection in failing hemodialysis access fistulas and grafts: comparison of color Doppler ultrasonography, contrast-enhanced magnetic resonance angiography, and digital subtraction angiography. AB - OBJECTIVE: Several imaging modalities are available for the evaluation of dysfunctional hemodialysis shunts. Color Doppler ultrasonography (CDUS) and digital subtraction angiography (DSA) are most widely used for the detection of access stenoses, and contrast-enhanced magnetic resonance angiography (CE-MRA) of shunts has recently been introduced. To date, no study has compared the value of these three modalities for stenosis detection in dysfunctional shunts. We prospectively compared CDUS and CE-MRA with DSA for the detection of significant (> or = 50%) stenoses in failing dialysis accesses, and we determined whether the interventionalist would benefit from CDUS performed before DSA and endovascular intervention. METHODS: CDUS, CE-MRA, and DSA were performed of 49 dysfunctional hemodialysis arteriovenous fistulas and 32 grafts. The vascular tree of the accesses was divided into three to eight segments depending on the access type (arteriovenous fistula or arteriovenous graft) and the length of venous outflow. CDUS was performed and assessed by a vascular technician, whereas CE-MRA and DSA were interpreted by two magnetic resonance radiologists and two interventional radiologists, respectively. All readers were blinded to information from each other and from other studies. DSA was used as reference standard for stenosis detection. RESULTS: DSA detected 111 significant (> or = 50%) stenoses in 433 vascular segments. Sensitivity and specificity of CDUS for the detection of significant stenosed vessel segments were 91% (95% CI, 84%-95%) and 97% (95% CI, 94%-98%), respectively. We found a positive predictive value of 91% (95% CI, 84% 95%) and a negative predictive value of 97% (95% CI, 94%-98%). The sensitivity, specificity, positive predictive value, and negative predictive value of MRA were 96% (95% CI, 90%-98%), 98% (95% CI, 96%-99%), 94% (95% CI, 88%-97%), and 98% (95% CI, 96%-99%), respectively. CDUS and CE-MRA depicted respectively three and four significant stenoses in six nondiagnostic DSA segments. The interventionalist would have chosen an alternative cannulation site in 38% of patients if the CDUS results had been available. CONCLUSIONS: We suggest that CDUS be used as initial imaging modality of dysfunctional shunts, but complete access should be depicted at DSA and angioplasty to detect all significant stenoses eligible for intervention. CE-MRA should be considered only if DSA is inconclusive. PMID- 16242564 TI - Pressure distention compared with pharmacologic relaxation in vein grafting upregulates matrix metalloproteinase-2 and -9. AB - OBJECTIVE: Autogenous vein bypasses are a common and effective method to treat occlusive disease. During surgical preparation, veins are routinely pressure distended to overcome vasospasm and twists. Distention, however, is believed to promote vascular remodeling and contribute to decreased graft patency. Pharmacologic vasorelaxation with a combination of effective vasodilators has been suggested as an alternative to pressure distention. The extracellular matrix (ECM)-degrading matrix metalloproteinases (MMPs) have been implicated in vascular remodeling and neointima formation. The purpose of the present study was to compare the effects of pressure distention with pharmacologic vasorelaxation on graft remodeling and regulation of MMP-2 and MMP-9 in porcine vein grafts. METHODS: Carotid artery bypass utilizing internal jugular veins was performed in eight female white pigs. Jugular veins were randomized to receive pressure distention (300 mm Hg for 2 minutes) or a combination of vasodilators (the alpha adrenergic antagonist phenoxybenzamine, 10 micromol/L; the Rho-kinase inhibitor HA-1077 [fasudil], 50 micromol/L; and the calcium-channel blocker nicardipine, 1 micromol/L) for 30 minutes and then were grafted into the carotid arteries. Two weeks after surgery, vein graft samples were analyzed for vessel intimal and medial area, lumen diameter, and ECM composition. Molecular analysis using reverse transcription-polymerase chain reaction, Western immunoblotting, gelatin zymography, and reverse zymography were performed to study the expression and activation of MMP-2 and MMP-9, and tissue inhibitors of MMP (TIMP)-1 and TIMP-2. RESULTS: Pressure distention irreversibly overstretched the porcine jugular vein and increased MMP-2 and MMP-9 proteolytic activity by 40% and 77%, respectively. Two weeks of vein grafting in the carotid arterial bed induced vessel wall thickening, ECM modification, and neointima formation, which were more pronounced in the distended grafts (P < .05) and accompanied by an increase in MMP expression and activity. Distended grafts demonstrated higher percentages of active MMP-9 (17.8% +/- 1.0%) and higher activities of latent (35.5% +/- 3.3%) and active MMP-2 (69.6% +/- 8.8%) than the pharmacologically treated grafts. Protein expression of TIMP-1 and TIMP-2 was downregulated after arterial grafting, but the pharmacologically treated grafts expressed significantly more TIMP-1 protein (by 36.8% +/- 4.1%) than the distended ones. The activities of TIMPs were markedly decreased after grafting, contributing to the upregulated MMP activity. CONCLUSIONS: Pressure distention of vein grafts before implantation, compared with pharmacologic vasodilatation, stimulates neointima formation and augments MMP activities. Pharmacologic vasorelaxation may be clinically superior to distention in attenuating graft remodeling and possibly improving graft patency. CLINICAL RELEVANCE: Autogenous vein bypasses are a common and effective method to treat occlusive disease. This study demonstrated that pressure distention, a common preparatory procedure in bypass surgery, upregulates extracellular matrix-degrading matrix metalloproteinases, which predisposes vein grafts to extensive remodeling and contributes to neointima formation and graft occlusion. The topical application of a combination of vasodilators to the vein graft before implantation may be clinically superior to pressure distention in attenuating graft remodeling and may possibly improve graft patency and reduce secondary surgical interventions. PMID- 16242565 TI - Hydrophilic statin suppresses vein graft intimal hyperplasia via endothelial cell tropic Rho-kinase inhibition. AB - BACKGROUND: Recent studies suggest that statins can protect the vasculature in a manner that is independent of their lipid-lowering activity through inhibition of the small guanosine triphosphate-binding protein, Rho, and Rho-associated kinase. Little information is available on the inhibitory effect of statins on vein graft intimal hyperplasia, the main cause of late graft failure after bypass grafting. We therefore examined the effects of a hydrophilic statin on vein graft intimal hyperplasia in vivo and Rho-kinase activity in vitro. METHODS: In the first experiment, rabbits were randomized to a control group (n = 7) that was fed regular rabbit chow or to a pravastatin group (n = 7) that was fed regular rabbit chow supplemented with 10 mg/kg pravastatin sodium. The branches of the jugular vein were ligated and an approximately 3-cm segment of the jugular vein was taken for an autologous reversed-vein graft. The carotid artery was cut and replaced with the harvested autologous jugular vein. At 2 and 4 weeks after the operation, vein grafts in both groups were harvested, and intimal hyperplasia of the vein grafts was assessed. In the second experiment, human umbilical vein endothelial cells and vascular smooth muscle cells were cultured and then treated with 1 micromol/L and 30 micromol/L pravastatin for 24 hours and harvested. Immunoblotting was performed on the resulting precipitates. Quantitative evaluation of phosphorylated myosin binding subunit and endothelial nitric oxide synthase was performed by densitometric analysis. RESULTS: We demonstrated that oral administration of the hydrophilic statin pravastatin to normocholesterolemic rabbits inhibited intimal hyperplasia of carotid interposition-reversed jugular vein grafts 4 weeks after implantation (pravastatin group, 39.5 +/- 3.5 microm vs control group, 64.0 +/- 7.1 microm; n = 7; P < .05) and suppressed cell proliferation and apoptosis in the neointima 2 weeks after implantation. In addition, we found that pravastatin inhibited Rho-kinase activity and accelerated endothelial nitric oxide synthase expression in human umbilical vein endothelial cells but did not inhibit Rho-kinase activity in vascular smooth muscle cells. CONCLUSIONS: These novel findings clearly demonstrate that a hydrophilic statin can suppress intimal hyperplasia of the vein graft in vivo and also show endothelial cell-tropic inhibition of Rho-kinase in vitro. Furthermore, these results strongly support the clinical use of hydrophilic statins to prevent intimal hyperplasia of the vein graft after bypass grafting. CLINICAL RELEVANCE: Late graft failure caused by neointimal hyperplasia limits the efficacy of vein grafting. Various treatments were examined to reduce neointimal hyperplasia, but a standard clinical treatment has not yet been established. We report here the inhibitory effect of pravastatin on the development of vein graft intimal hyperplasia. In addition, we demonstrate that pravastatin showed endothelial cell tropic benefits through both the inhibition of Rho-kinase activity and acceleration of eNOS expression in vitro. Because the clinical benefits and safety of pravastatin have been established to a certain extent through long-term clinical usage, pravastatin may soon become standard treatment after vein bypass grafting. PMID- 16242566 TI - Protective use of N-methyl-D-aspartate receptor antagonists as a spinoplegia against excitatory amino acid neurotoxicity. AB - OBJECTIVE: Paraplegia remains a serious complication of thoracic and thoracoabdominal aortic operations. To avoid this dreadful complication, N-methyl D-aspartate (NMDA) receptor antagonists have been examined in the ischemic or excitotoxic neuronal injury model. In the present study, we evaluated the protective efficacy of NMDA receptor antagonists that were infused segmentally after aortic clamping, as a spinoplegia, to reduce aspartate neurotoxicity in the spinal cord. METHODS: Infrarenal aortic isolation was performed in New Zealand white rabbits. Group A animals (n = 7) were pretreated with the segmental infusion of MK-801, a noncompetitive NMDA receptor antagonist, followed by segmental aspartate (50 mmol) infusion for 10 minutes. Group B animals (n = 6) received pretreatment with CGS19755, a competitive NMDA receptor antagonist, followed by the same aspartate infusion as group A. Group C animals (n = 7) received vehicle only, followed by aspartate infusion as a control group. In addition, group D animals (n = 6) were pretreated with MK-801 that was administrated intravenously 1 hour before aspartate infusion. Neurologic status was assessed at 12, 24, and 48 hours after operation by using the Tarlov score. The spinal cords were procured at 48 hours for histopathologic analysis to determine the extent of excitotoxic neuronal injury. RESULTS: Most of the animals in groups A and D revealed full recovery or mild motor disturbance. Group B and C animals exhibited paraplegia or paraparesis with marked neuronal necrosis. In the Tarlov score at 48 hours, group A animals represented better neurologic function than group C (P < .01) and similar motor function to group D animals. Severe histopathologic change was not observed in groups A and D. Animals in groups A and D showed a greater number of motor neurons than animals in groups B and C (P < .01). The difference could be due to chance between group A and D animals (P = .08). CONCLUSIONS: These results showed that the segmental infusion of noncompetitive NMDA receptor antagonist as an intraoperative spinoplegia could have a protective effect on the spinal cord neurons against excitotoxic neuronal injury in vivo. On the other hand, efficacy of the use of competitive antagonist was suggested to be limited in this model, probably because of the insurmountable obstacle of the blood-brain barrier. CLINICAL RELEVANCE: Paraplegia is a devastating complication during surgical repair of the thoracic and thoracoabdominal aortas. Excitatory amino acids neurotoxicity through the N methyl-D-aspartate (NMDA) receptor is no doubt the pathologic hallmark of ischemic and postischemic spinal cord injury. Systemic administration of either a competitive or noncompetitive NMDA antagonist has been reported to have neuroprotective effect, in terms of preoperative treatment, with dose-related central sympathomimetic and sedative effects. Local administration, particularly of a noncompetitive NMDA antagonist, infused segmentally after aortic clamping could therefore be a potent intraoperative pharmacologic strategy to minimize the effective dose that retains NMDA antagonism without undesirable adverse effects. Our ability to reproduce this model could facilitate pharmacologic prevention or provide a new surgical technique as a spinoplegia for NMDA receptor-mediated neuronal injury. PMID- 16242568 TI - Endovascular stent graft for symptomatic mobile thrombus of the thoracic aorta. AB - Mobile thoracic aortic thrombus is a potential source of arterial embolism. Therapeutic management remains controversial. Systemic anticoagulation and various open surgical procedures are the commonly used therapeutic modalities. We report the successful primary treatment by endovascular stent graft of a mobile thoracic aortic thrombus that had caused visceral and peripheral embolism. Our case shows that endovascular stent-graft treatment is an effective, minimally invasive treatment of symptomatic mobile thoracic aortic thrombus. PMID- 16242567 TI - Sustained orbital shear stress stimulates smooth muscle cell proliferation via the extracellular signal-regulated protein kinase 1/2 pathway. AB - OBJECTIVE: Nonlaminar shear stress stimulates smooth muscle cell (SMC) proliferation and migration in vivo, especially after an endothelial-denuding injury. To determine whether sustained shear stress directly stimulates SMC proliferation in vitro, the effect of orbital shear stress on SMC proliferation, phenotype, and extracellular signal-regulated protein kinase 1/2 (ERK1/2) phosphorylation was examined. METHODS: Bovine SMCs were exposed to orbital shear stress (210 rpm) for up to 10 days, with and without the ERK1/2 upstream pathway inhibitor PD98059 (10 microM) or the p38 pathway inhibitor SB203580 (10 microM). Proliferation was directly counted and assessed with proliferation cell nuclear antigen. Western blotting was used to assess activation of SMC ERK1/2 and SMC phenotype markers. RESULTS: SMCs exposed to sustained orbital shear stress (10 days) had 75% increased proliferation after 10 days compared with static conditions. Expression of markers of the contractile phenotype (alpha-actin, calponin) was decreased, and markers of the synthetic phenotype (vimentin, beta actin) were increased. ERK1/2 was phosphorylated in the presence of orbital shear stress, and orbital shear-stress-stimulated SMC proliferation was inhibited in the presence of PD98059 but sustained in the presence of SB203580. Orbital shear stress-induced changes in SMC phenotype were also inhibited in the presence of PD98059. CONCLUSION: Orbital shear stress directly stimulates SMC proliferation in long-term culture in vitro and is mediated, at least partially, by the ERK1/2 pathway. The ERK1/2 pathway may also mediate the orbital shear-stress-stimulated switch from SMC contractile to synthetic phenotype. These results suggest that shear-stress-stimulated SMC proliferation after vascular injury is mediated by a pathway amenable to pharmacologic manipulation. PMID- 16242569 TI - Retrograde migration of an abdominal aortic aneurysm endograft leading to postoperative renal failure. AB - The phenomenon of antegrade (distal) endograft migration is an extensively documented complication after endovascular abdominal aortic aneurysm repair. This case report describes the finding of postoperative abdominal aortic aneurysm endograft retrograde (proximal) migration occluding bilateral renal arteries and leading to dialysis-dependent renal failure. PMID- 16242570 TI - Superficial femoral artery transposition repair for isolated superior mesenteric artery dissection. AB - Isolated dissection of the superior mesenteric artery is an uncommon event, but many new cases have been reported recently, reflecting the progress of imaging and suggesting that this pathology is not as rare as previously thought. Here we report a case of superior mesenteric artery dissection where we performed, after failure of conservative medical management, an original surgical technique for mesenteric revascularization using a superficial femoral artery transposition. To the best of our knowledge, this is the first report of the use of this technique for complex mesenteric revascularization. PMID- 16242571 TI - Cerebral venous hypertension and blindness: a reversible complication. AB - A 57-year-old woman developed blindness during treatment for sarcoidosis-induced end-stage renal disease. An initial renal transplantation failed, and hemoaccess was maintained with multiple central catheters and upper extremity prosthetic arteriovenous grafts. A successful second transplantation eliminated her need for hemodialysis, but a right brachial to internal jugular graft remained patent. Progressive visual loss 2 years after transplantation prompted ophthalmic evaluation which initially revealed unilateral left optic nerve edema and visual loss, ultimately worsening over several months to no light perception in the left eye, 20/60 vision in the right eye, and bilateral papilledema. Arteriography demonstrated cerebral venous hypertension attributed to the functioning hemoaccess graft. Permanent graft occlusion normalized the papilledema, and visual field defects in the right eye and visual acuity returned to 20/20 in the right eye. PMID- 16242572 TI - Brachial artery occlusion secondary to hypereosinophilic syndrome. AB - Hypereosinophilic syndrome (HES) is characterized by an overproduction of eosinophils that leads to organ damage. Although most cases of HES frequently affect the lungs, heart, and gastrointestinal tract, there are a few reported cases of peripheral vascular involvement. We report a case of a patient with a history of colonic HES who presented with idiopathic occlusion of the brachial artery. A 28-year-old woman with a recent history of eosinophilic colitis presented with a several-week history of left hand pain, pallor, and paresthesias. Her hand was cool, without palpable pulses. Her eosinophilia count was 38%. An arteriogram documented a left brachial artery occlusion and diffuse left arm vasospasm. A brachial-to-brachial bypass was performed. Postoperatively, there was extensive vasospasm of her distal upper extremity arteries, which was treated with calcium-channel blockers and steroids. Her symptoms resolved and she has been asymptomatic for 9 months. The segment of occluded artery was found to contain many eosinophils on histologic examination. HES of the arterial system is an exceedingly rare cause of occlusion. Our patient presented with eosinophilia, arterial vasospasm with subsequent occlusion, and the presence of eosinophilic infiltration on the pathologic specimen. These data, combined with the patient's previous history, demonstrate that the patient's occlusion may have been secondary to HES. PMID- 16242573 TI - Wegener's granulomatosis presenting during pregnancy with acute limb ischemia. AB - We report a rare case of a pregnant woman with Wegener's granulomatosis whose disease involved the lungs, the spleen, and a limb. Wegener's granulomatosis was clinically manifested in the 34th week of pregnancy with pulmonary infiltrates, splenic infarcts, and acute limb ischemia. Successive thrombectomies failed to maintain arterial flow in the distal limb due to the development of active vasculitis. Thrombosis of the tibial arteries and recurrence of thrombosis was a persistent clinical observation. In the meantime, a cesarean section was performed, with a successful delivery of a healthy male newborn. Because of the recent cesarean section, the patient did not undergo thrombolysis. The woman finally underwent amputation of the limb. According to the reviewed literature, this is the sixth reported case of Wegener's granulomatosis presenting with digital ischemia and the first manifested during pregnancy. PMID- 16242574 TI - Secondary aortoenteric fistula. PMID- 16242575 TI - The two-stage brachial artery-brachial vein autogenous fistula for hemodialysis: an alternative autogenous option for hemodialysis access. AB - The optimal dialysis access for the patient with chronic renal failure is considered to be an autogenous fistula; this is reflected in the recommendations of the National Kidney Foundation-Disease Outcomes Quality Initiatives (NKF DOQI). If adequate superficial veins at the wrist or the forearm are not available, the next option is usually a prosthetic arteriovenous graft. In this case series, we describe our experience with an autogenous fistula constructed using the brachial vein. There were 20 patients over a 14-month period who were operated on for dialysis access. In these patients, no adequate superficial veins were found at operation. Instead of using a prosthetic graft, we performed a brachial artery-brachial vein fistula in two stages. The first stage involved a forearm anastomosis and then subsequently, weeks later, this fistula was "superficialized." Twenty patients underwent a brachial artery-brachial vein fistula. Of these patients, all had successful maturation of their fistula and after a minimum waiting period of 12 weeks for maturation; all but one were able to be successfully dialyzed through their fistula. One patient developed arm swelling due to previously placed subclavian vein pacemaker wires. None of the other patients developed arm swelling or vascular steal. The brachial artery brachial vein fistula is a feasible option for hemodialysis access and we suggest that this option be considered before a prosthetic arteriovenous graft is inserted. Arm swelling and steal have not been a problem, and all patients have been able to have full dialysis through the fistula after appropriate maturation times. PMID- 16242576 TI - Vascular surgery: a specialty in transformation with a bright future. PMID- 16242577 TI - The ethics of bylines: would the real authors please stand up? PMID- 16242578 TI - Regarding "Noncontrast three-dimensional magnetic resonance imaging vs lymphoscintigraphy in the evaluation of lymph circulation disorders: a comparative study". PMID- 16242581 TI - Regarding "Percutaneous angioplasty and stenting of the superficial femoral artery". PMID- 16242582 TI - Regarding: "The study of endovascular repair of small (<5.5-cm) aneurysms". PMID- 16242583 TI - Regarding "Perioperative beta-blockade (POBBLE) for patients undergoing infrarenal vascular surgery: results of a randomized double-blind controlled trial". PMID- 16242586 TI - Randomized controlled trial of physical activity counseling for older primary care patients. AB - BACKGROUND: Regular physical activity reduces the risk for chronic diseases among older adults. Older adults are likely to be seen by primary care clinicians who can play a role in promoting physical activity among their patients. DESIGN: In this randomized controlled trial (1998-2003; data analyzed 2004-2005), we compared the effects of brief advice to exercise from a clinician supplemented by telephone-based counseling by health educators (extended advice) to brief advice from a clinician alone (brief advice). SETTING/PARTICIPANTS: A total of 100 primary care patients (63.2% female, 14.7% minority, mean age=68.5 years) participated in the trial. INTERVENTIONS: The extended-advice intervention consisted of clinician advice plus exercise counseling via telephone provided by research staff, and the brief advice condition consisted of clinician advice alone. Both interventions focused on promoting moderate-intensity physical activity. MAIN OUTCOME MEASURES: Self-reported physical activity using the 7-Day Physical Activity Recall instrument and objective activity monitoring using Biotrainers were assessed at baseline, and at 3 and 6 months. RESULTS: Participants in the extended-advice arm reported significantly greater participation in moderate-intensity physical activity than the brief-advice group at 3 months (+57.69 minutes vs 12.45 minutes; 3.84 kcal/week vs 0.83 kcal/week) and 6 months (+62.84 minutes vs 16.60 minutes; 4.19 kcal/week vs 1.1 kcal/week). Objective activity monitoring also showed significantly increased physical activity among extended-advice versus brief-advice participants at both time points (+50.79 vs -11.11; +42.39 vs -24.18, respectively). CONCLUSIONS: These data indicate that clinician advice with follow-up counseling can promote adoption of moderate-intensity physical activity among older, primary care patients. PMID- 16242587 TI - Screening for physical activity in family practice: evaluation of two brief assessment tools. AB - BACKGROUND: Physical activity (PA) is relevant to the prevention and management of many health conditions in family practice. There is a need for an efficient, reliable, and valid assessment tool to identify patients in need of PA interventions. METHODS: Twenty-eight family physicians in three Australian cities assessed the PA of their adult patients during 2004 using either a two- (2Q) or three-question (3Q) assessment. This was administered again approximately 3 days later to evaluate test-retest reliability. Concurrent validity was evaluated by measuring agreement with the Active Australia Questionnaire, and criterion validity by comparison with 7-day Computer Science Applications, Inc. (CSA) accelerometer counts. RESULTS: A total of 509 patients participated, with 428 (84%) completing a repeat assessment, and 415 (82%) accelerometer monitoring. The brief assessments had moderate test-retest reliability (2Q k=58.0%, 95% confidence interval [CI]=47.2-68.8%; 3Q k=55.6%, 95% CI=43.8-67.4%); fair to moderate concurrent validity (2Q k=46.7%, 95% CI=35.6-57.9%; 3Q k=38.7%, 95% CI=26.4-51.1%); and poor to fair criterion validity (2Q k=18.2%, 95% CI=3.9 32.6%; 3Q k=24.3%, 95% CI=11.6-36.9%) for identifying patients as sufficiently active. A four-level scale of PA derived from the PA assessments was significantly correlated with accelerometer minutes (2Q rho=0.39, 95% CI=0.28 0.49; 3Q rho=0.31, 95% CI=0.18-0.43). Physicians reported that the assessments took 1 to 2 minutes to complete. CONCLUSIONS: Both PA assessments were feasible to use in family practice, and were suitable for identifying the least active patients. The 2Q assessment was preferred by clinicians and may be most appropriate for dissemination. PMID- 16242588 TI - Prospective study of physical activity and depressive symptoms in middle-aged women. AB - BACKGROUND: Although many studies support an inverse association between physical activity (PA) and depressive symptoms, prospective relationships between these variables have been confounded by pre-existing psychological and physical health problems. METHODS: This study examined the dose-response relationships between self-reported PA and depressive symptoms, using cross-sectional and prospective data from a population-based cohort of middle-aged women who participated in the Australian Longitudinal Study on Women's Health (ALSWH) between 1996 and 2001. Participants completed three mailed surveys (S1, 1996; S2, 1998; S3, 2001), which included questions about time spent in walking, moderate- and vigorous-intensity PA, and measures of psychological health (Center for Epidemiologic Studies Depression scale [CESD-10], and Mental health [MH] subscale of the Short Form 36 survey). Relationships between previous (S1, S2), current (S3), and habitual (S1, S2, S3) PA and "depressive symptoms" were examined, adjusting for sociodemographic and health-related variables (n=9207). RESULTS: Mean CESD-10 scores decreased, and MH scores increased with increasing levels of previous, current, and habitual activity. Odds ratios for CESD-10 scores > or =10 or MH scores < or =52 at S3 were 30% to 40% lower among women who reported the equivalent of > or =60 minutes of moderate-intensity PA per week, compared with those who reported less PA than this. Women who were in the lowest PA category at S1, but who subsequently reported > or =240 metabolic equivalent minutes (MET.mins) per week had lower odds of CESD-10 scores of > or =10 or MH scores < or =52 at S3 than those who remained in the very low PA category. CONCLUSIONS: These data suggest that there is a clear relationship between increasing PA and decreasing depressive symptoms in middle-aged women, independent of pre-existing physical and psychological health. PMID- 16242589 TI - Walking to public transit: steps to help meet physical activity recommendations. AB - BACKGROUND: Nearly half of Americans do not meet the Surgeon General's recommendation of > or =30 minutes of physical activity daily. Some transit users may achieve 30 minutes of physical activity daily solely by walking to and from transit. This study estimates the total daily time spent walking to and from transit and the predictors of achieving 30 minutes of physical activity daily by doing so. METHODS: Transit-associated walking times for 3312 transit users were examined among the 105,942 adult respondents to the 2001 National Household Travel Survey, a telephone-based survey sponsored by the U.S. Department of Transportation to assess American travel behavior. RESULTS: Americans who use transit spend a median of 19 minutes daily walking to and from transit; 29% achieve > or =30 minutes of physical activity a day solely by walking to and from transit. In multivariate analysis, rail users, minorities, people in households earning <$15,000 a year, and people in high-density urban areas were more likely to spend > or =30 minutes walking to and from transit daily. CONCLUSIONS: Walking to and from public transportation can help physically inactive populations, especially low-income and minority groups, attain the recommended level of daily physical activity. Increased access to public transit may help promote and maintain active lifestyles. Results from this study may contribute to health impact assessment studies (HIA) that evaluate the impact of proposed public transit systems on physical activity levels, and thereby may influence choices made by transportation planners. PMID- 16242590 TI - Using ecologic momentary assessment to measure physical activity during adolescence. AB - OBJECTIVE: Examine the validity of using high-density electronic ecologic momentary assessment (EMA) to assess physical activity. EMA was further used to explore within- and between-subject variability in adolescent physical activity (PA) patterns. METHODS: Adolescents (n=526, 51% male) participated in EMA waves occurring approximately every 6 months between the 9th and 12th grade. Each wave extended over 4 consecutive days (Thursday to Sunday). Using a Palm III handheld computer, each participant reported his or her primary activity (e.g., exercise, walking, homework) every 30 (+/-10) minutes during waking hours. Heart rate (via Polar heart rate monitor) and activity counts (via wrist accelerometer) were simultaneously assessed during the EMA intervals. RESULTS: Overall, heart rates and accelerometer counts were greater for diary-reported exercise and walking than for nonphysical activities (p's<0.001). EMA revealed that the typical duration of exercise sessions was longer than walking sessions (p<0.05). Rates of walking and exercise were more consistent between waves (i.e., across high school) than within waves (i.e., across the 4 days of monitoring), most likely due to the significantly higher rates of walking and exercise occurring on weekdays as compared to weekend days (p's<0.001). Average rates of walking were greater for girls than boys, and the reverse was true for exercise. Rates of both walking and exercise declined steadily between 9th and 12th grade (p's<0.001). CONCLUSIONS: Physical activity reported via EMA corresponded to objective activity indicators. EMA yielded information about within-person variability in PA that cannot be obtained readily from traditional self-report instruments. Given its potential for simultaneously assessing important physiologic, psychological, and contextual factors, EMA presents a promising approach to studying adolescent physical activity. PMID- 16242591 TI - Use and effectiveness of tobacco telephone counseling and nicotine therapy in Maine. AB - BACKGROUND: Since 2001, the Maine Bureau of Health has offered free evidence based treatment for tobacco dependence, including telephonic counseling and nicotine replacement therapy (NRT). This study examined the utilization of treatment services, evaluated quit outcomes, and estimated the population impact of treatment. METHODS: This is a descriptive study of tobacco users receiving treatment services from the Maine Tobacco HelpLine from January 2003 to December 2004. Demographics of callers were compared to adult smokers statewide, and NRT utilization was examined among callers eligible for therapy. Quit outcomes were assessed by telephone interview among a sample of callers registered November 15, 2003 to January 31, 2004 (n=535), 6 months after assistance. The population impact of treatment was estimated by applying intent-to-treat (30-day point prevalence) quit rates to services delivered in 2003 and 2004. Analyses were conducted in 2005. RESULTS: A total of 12,479 adult smokers (3% of smokers annually) utilized Maine's tobacco services during 2003 and 2004. Compared to smokers statewide, callers were more likely to be aged 45 to 64, female, or uninsured. A total of 82.3% of callers who were eligible for NRT and received counseling obtained free NRT. Intent-to-treat quit rates at 6 months were 12.3% (95% confidence interval [CI]=8.1-17.6) for counseling, and 22.5% (95% CI=19.1 26.3) for counseling plus NRT. An estimated 1864 smokers calling in 2003-2004 had successfully quit. CONCLUSIONS: The Maine Tobacco HelpLine and NRT programs have demonstrated effectiveness and population outreach, particularly to uninsured smokers. This study suggests that for quit lines to maximize their impact, tobacco medication access may be important. PMID- 16242592 TI - Incidence of TB in inmates with latent TB infection: 5-year follow-up. AB - BACKGROUND: Inmates are a high-risk population for tuberculosis (TB) control efforts, including treatment for latent tuberculosis infection (LTBI). Completion of therapy after release has been poor. The goal of this study was to evaluate therapy completion and active disease over 5 years in a cohort of inmates. METHODS: The sample was from a completed randomized trial in 1998-1999 of education or incentive versus usual care to improve therapy completion after release from the San Francisco County Jail. Records from the jail, the County Tuberculosis Clinic, and the California TB Registry were used to measure therapy completion and development of active TB. Analyses were conducted in 2005. RESULTS: Of a total 527 inmates, 31.6% (n=176) completed therapy, of whom 59.7% (n=105) completed it in jail. Compared with the U.S.-born, foreign-born inmates residing in the United States for < or =5 years were less likely to complete the therapy (adjusted odds ratio [AOR]=0.49, 95% confidence interval [CI]=0.28-0.85), and those with more education were more likely to complete the therapy (AOR=1.06, 95% CI=1.01-1.12). Three subjects developed active TB in the 5 years of follow up, resulting in an annual rate of 108 per 100,000. Compared with California rates, subjects were 59 times as likely to develop active TB (standardized morbidity ratio of 59.2, 95% CI=11.2-145.1). None had completed therapy, none were new immigrants, and two were known to be HIV-positive at diagnosis. CONCLUSIONS: Completion of therapy for LTBI is a challenge, but the active TB seen in this jail cohort emphasizes the importance of continued efforts to address TB risk in this population. PMID- 16242593 TI - Prevention of upper respiratory tract infections by gargling: a randomized trial. AB - BACKGROUND: Gargling to wash the throat is commonly performed in Japan, and people believe that such hygienic routine, especially with gargle medicine, prevents upper respiratory tract infections (URTIs). Its effectiveness, however, has not been established by clinical trials. DESIGN: Randomized controlled trial carried out in 2002-2003 winter season and analyzed in 2003 and 2004. PARTICIPANTS: Healthy volunteers (387) aged 18 to 65 years. INTERVENTION: Participants were randomly assigned to water gargling, povidone-iodine gargling, and usual care (control). Subjects in the two gargling groups were requested to gargle with water or diluted povidone-iodine at least three times a day. Participants were followed for 60 days. MAIN OUTCOME MEASURES: The primary outcome measure was first URTI incidence. Severity of URTI symptoms among incident cases was also evaluated. Both outcomes were assessed with a self administered symptom record. Analyses were performed on an intention-to-treat basis. RESULTS: A total of 130 participants contracted URTIs. The incidence rate of first URTI was 0.26 episodes/30 person-days among control subjects. The rate decreased to 0.17 episodes/30 person-days in the water gargling group, and 0.24 episodes/30 person-days in the povidone-iodine gargling group. Respective incidence rate ratios against controls were 0.64 (95% confidence interval [CI]=0.41-0.99) and 0.89 (95% CI=0.60-1.33). A Cox regression (proportional hazard model) revealed the efficacy of water gargling (hazard ratio=0.60, 95% CI=0.39-0.95). Even when a URTI occurred, water gargling tended to attenuate bronchial symptoms (p=0.055). CONCLUSIONS: Simple water gargling was effective to prevent URTIs among healthy people. This virtually cost-free modality would appreciably benefit the general population. PMID- 16242594 TI - McDonald's restaurants and neighborhood deprivation in Scotland and England. AB - BACKGROUND: Features of the local fast food environment have been hypothesized to contribute to the greater prevalence of obesity in deprived neighborhoods. However, few studies have investigated whether fast food outlets are more likely to be found in poorer areas, and those that have are local case studies. In this paper, using national-level data, we examine the association between neighborhood deprivation and the density of McDonald's restaurants in small census areas (neighborhoods) in Scotland and England. METHODS: Data on population, deprivation, and the location of McDonald's Restaurants were obtained for 38,987 small areas in Scotland and England (6505 "data zones" in Scotland, and 32,482 "super output areas" in England) in January 2005. Measures of McDonald's restaurants per 1000 people for each area were calculated, and areas were divided into quintiles of deprivation. Associations between neighborhood deprivation and outlet density were examined during February 2005, using one-way analysis of variance in Scotland, England, and both countries combined. RESULTS: Statistically significant positive associations were found between neighborhood deprivation and the mean number of McDonald's outlets per 1000 people for Scotland (p<0.001), England (p<0.001), and both countries combined (p<0.001). These associations were broadly linear with greater mean numbers of outlets per 1000 people occurring as deprivation levels increased. CONCLUSIONS: Observed associations between presence or absence of fast food outlets and neighborhood deprivation may provide support for environmental explanations for the higher prevalence of obesity in poor neighborhoods. PMID- 16242595 TI - Interventions to improve the health of the homeless: a systematic review. AB - BACKGROUND: Homelessness is a widespread problem in the United States. The primary goal of this systematic review is to provide guidance in the development and organization of programs to improve the health of homeless people. METHODS: MEDLINE, CINAHL, HealthStar, PsycINFO, Sociological Abstracts, and Social Services Abstracts databases were searched from their inception through July 2004 using the following terms: homeless, homeless persons, and homelessness. References of key articles were also searched. 4564 abstracts were screened, and 258 articles underwent full review. Seventy-three studies conducted from 1988 to 2004 met inclusion criteria (use of an intervention, use of a comparison group, and the reporting of health-related outcomes). Two authors independently abstracted data from studies and assigned quality ratings using explicit criteria. RESULTS: Forty-five studies were rated good or fair quality. For homeless people with mental illness, case management linked to other services was effective in improving psychiatric symptoms, and assertive case management was effective in decreasing psychiatric hospitalizations and increasing outpatient contacts. For homeless people with substance abuse problems, case management resulted in greater decreases in substance use than did usual care. For homeless people with latent tuberculosis, monetary incentives improved adherence rates. Although a number of studies comparing an intervention to usual care were positive, studies comparing two interventions frequently found no significant difference in outcomes. CONCLUSIONS: Coordinated treatment programs for homeless adults with mental illness or substance abuse usually result in better health outcomes than usual care. Health care for homeless people should be provided through such programs whenever possible. Research is lacking on interventions for youths, families, and conditions other than mental illness or substance abuse. PMID- 16242596 TI - Fish: balancing health risks and benefits. PMID- 16242597 TI - Fish, health, and sustainability. PMID- 16242598 TI - Health trade-offs from policies to alter fish consumption. PMID- 16242599 TI - A quantitative risk-benefit analysis of changes in population fish consumption. AB - Although a rich source of n-3 polyunsaturated fatty acids (PUFAs) that may confer multiple health benefits, some fish contain methyl mercury (MeHg), which may harm the developing fetus. U.S. government recommendations for women of childbearing age are to modify consumption of high-MeHg fish, while recommendations encourage fish consumption among the general population because of nutritional benefits. To investigate the aggregate impacts of hypothetical shifts in fish consumption, the Harvard Center for Risk Analysis convened an expert panel (see acknowledgements). Effects investigated include prenatal cognitive development, coronary heart disease mortality, and stroke. Substitution of fish with high MeHg concentrations with fish containing less MeHg among women of childbearing age yields substantial developmental benefits and few negative impacts. However, if women instead decrease fish consumption, countervailing risks substantially reduce net benefits. If other adults (mistakenly and inappropriately) also reduce their fish consumption, the net public health impact is negative. Although high compliance with recommended fish consumption patterns can improve public health, unintended shifts in consumption can lead to public health losses. Risk managers should investigate and carefully consider how populations will respond to interventions, how those responses will influence nutrient intake and contaminant exposure, and how these changes will affect aggregate public health. PMID- 16242600 TI - A quantitative analysis of fish consumption and coronary heart disease mortality. AB - Although a rich source of n-3 polyunsaturated fatty acids (PUFAs) that may confer multiple health benefits, some fish contain methyl mercury (MeHg), which may harm the developing fetus. U.S. government recommendations for women of childbearing age are to modify consumption of high-MeHg fish to reduce MeHg exposure, while recommendations encourage fish consumption among the general population because of the nutritional benefits. The Harvard Center for Risk Analysis convened an expert panel (see acknowledgements) to quantify the net impact of resulting hypothetical changes in fish consumption across the population. This paper estimates the impact of fish consumption on coronary heart disease (CHD) mortality and nonfatal myocardial infarction (MI). Other papers quantify stroke risk and the impacts of both prenatal MeHg exposure and maternal intake of n-3 PUFAs on cognitive development. This analysis identified articles in a recent qualitative review appropriate for the development of a dose-response relationship. Studies had to satisfy quality criteria, quantify fish intake, and report the precision of the relative risk estimates. Relative risk results were averaged, weighted proportionately by precision. CHD risks associated with MeHg exposure were reviewed qualitatively because the available literature was judged inadequate for quantitative analysis. Eight studies were identified (29 exposure groups). Our analysis estimated that consuming small quantities of fish is associated with a 17% reduction in CHD mortality risk, with each additional serving per week associated with a further reduction in this risk of 3.9%. Small quantities of fish consumption were associated with risk reductions in nonfatal MI risk by 27%, but additional fish consumption conferred no incremental benefits. PMID- 16242601 TI - A quantitative analysis of fish consumption and stroke risk. AB - Although a rich source of n-3 polyunsaturated fatty acids (PUFAs) that may confer multiple health benefits, some fish contain methyl mercury (MeHg), which may harm the developing fetus. U.S. government recommendations for women of childbearing age are to modify consumption of high-MeHg fish to reduce MeHg exposure, while recommendations encourage fish consumption among the general population because of the nutritional benefits. The Harvard Center for Risk Analysis convened an expert panel (see acknowledgements) to quantify the net impact of resulting hypothetical changes in fish consumption across the population. This paper estimates the impact of fish consumption on stroke risk. Other papers quantify coronary heart disease mortality risk and the impacts of both prenatal MeHg exposure and maternal intake of n-3 PUFAs on cognitive development. This analysis identified articles in a recent qualitative literature review that are appropriate for the development of a dose-response relationship between fish consumption and stroke risk. Studies had to satisfy quality criteria, quantify fish intake, and report the precision of the relative risk estimates. The analysis combined the relative risk results, weighting each proportionately to its precision. Six studies were identified as appropriate for inclusion in this analysis, including five prospective cohort studies and one case-control study (total of 24 exposure groups). Our analysis indicates that any fish consumption confers substantial relative risk reduction compared to no fish consumption (12% for the linear model), with the possibility that additional consumption confers incremental benefits (central estimate of 2.0% per serving per week). PMID- 16242602 TI - A quantitative analysis of prenatal methyl mercury exposure and cognitive development. AB - Although a rich source of n-3 polyunsaturated fatty acids (PUFAs) that may confer multiple health benefits, some fish also contain methyl mercury (MeHg), which may harm the developing fetus. U.S. government recommendations for women of childbearing age are to modify consumption of high MeHg fish to reduce MeHg exposure, while recommendations encourage fish consumption among the general population because of the nutritional benefits. The Harvard Center for Risk Analysis convened an expert panel (see acknowledgements) to quantify the net impact of resulting hypothetical changes in fish consumption across the population. This paper quantifies the impact of prenatal MeHg exposure on cognitive development. Other papers quantify the beneficial impact of prenatal intake of n-3 PUFAs on cognitive function and the extent to which fish consumption protects against coronary heart disease mortality and stroke in adults. This analysis aggregates results from three major prospective epidemiology studies to quantify the association between prenatal MeHg exposure and cognitive development as measured by intelligence quotient (IQ). It finds that prenatal MeHg exposure sufficient to increase the concentration of mercury in maternal hair at parturition by 1 microg/g decreases IQ by 0.7 points. This paper identifies important sources of uncertainty influencing this estimate, concluding that the plausible range of values for this loss is 0 to 1.5 IQ points. PMID- 16242603 TI - A quantitative analysis of prenatal intake of n-3 polyunsaturated fatty acids and cognitive development. AB - Although a rich source of n-3 polyunsaturated fatty acids (PUFAs) that may confer multiple health benefits, some fish also contain methyl mercury (MeHg), which may harm the developing fetus. U.S. government recommendations for women of childbearing age are to modify consumption of high-MeHg fish to reduce MeHg exposure, while recommendations encourage fish consumption among the general population because of the nutritional benefits. The Harvard Center for Risk Analysis convened an expert panel (see acknowledgements) to quantify the net impact of resulting hypothetical changes in fish consumption across the population. This paper estimates the impact of prenatal n-3 intake on cognitive development. Other papers quantify the negative impact of prenatal exposure to MeHg on cognitive development, and the extent to which fish consumption protects against coronary heart disease mortality and stroke in adults. This paper aggregates eight randomized controlled trials (RCTs) comparing cognitive development in controls and in children who had received n-3 PUFA supplementation (seven studies of formula supplementation and one study of maternal dietary supplementation). Our analysis assigns study weights accounting for statistical precision, relevance of three endpoint domains (general intelligence, verbal ability, and motor skills) to prediction of IQ, and age at evaluation. The study estimates that increasing maternal docosahexaenoic acid (DHA) intake by 100 mg/day increases child IQ by 0.13 points. The paper notes that findings were inconsistent across the RCTs evaluated (although our findings were relatively robust to changes in the weighting scheme used). Also, for seven of the eight studies reviewed, effects are extrapolated from formula supplementation to maternal dietary intake. PMID- 16242604 TI - A definition and operational framework for health numeracy. AB - Health numeracy has often been overshadowed by health literacy, either ignored completely or identified simply as a subset of health literacy. Only now are researchers beginning to realize the importance of health numeracy as a separate entity. One of the first steps in this evolution is to establish a distinct definition for health numeracy, something that has not been addressed in the literature to date. This paper proposes such a definition, as well as a set of clarifying categories in hopes of helping researchers both to advance the field of health numeracy and to focus their topics within the realm of health numeracy. PMID- 16242605 TI - The cost effectiveness of health insurance. PMID- 16242607 TI - On-chip micro-biosensor for the detection of human CD4(+) cells based on AC impedance and optical analysis. AB - The current study was undertaken to fabricate a small micro-electrode on-chip to rapidly detect and quantify human CD4(+) cells in a minimal volume of blood through impedance measurements made with simple electronics that could be battery operated implemented in a hand held device. The micro-electrode surface was non covalently modified sequentially by incubation with solutions of protein G', human albumin, monoclonal mouse anti-human CD4, and mouse IgG. The anti-human CD4 antibody served as the recognition and capture molecule for CD4(+) cells present in human blood. The binding of these biomolecules to the micro-electrodes was verified by impedance and cyclic voltammetry measurements. An increase in impedance was detected for each layer of protein adsorbed onto the micro electrode surface. This process was shown to be highly repeatable. Increased impedance was measured when CD4(+) cells were captured on the micro-electrode, and the impedance also increased as the number of captured cells increased. Fluorescence microscopy of captured cells immunolabeled with anti-human CD4, CD8, and CD19 antibodies, and the nuclear label DAPI, confirmed that only CD4(+) cells were captured. The results were highly dependent on the specimen preparation method used. We conclude that the on-chip capture system can efficiently quantify the number of CD4(+) cells. PMID- 16242608 TI - Investigation and evaluation of a method for determination of ethanol with the SIRE Biosensor P100, using alcohol dehydrogenase as recognition element. AB - A new method for rapid determination of ethanol was developed, using alcohol dehydrogenase as recognition element for the SIRE (sensors based on injection of the recognition element) Biosensor, which is an amperometric biosensor. The method was simple, fast, accurate, specific and cost-effective. The recognition element solution used was stable at least for 24 h in room temperature, and at least one month when lyophilised. The optimal potential versus the silver wire electrode, the optimal pH of the buffer and the optimal temperature of the water bath was determined to be +950 mV, 8.1 and 308 K, respectively. The optimal concentrations of alcohol dehydrogenase, BSA and NAD(+) were determined to be 200 U/ml, 20 mg/ml and 15 mM, respectively. The total analysis time was between 50 s and 4 min per analysis, depending on the concentration range. The linear range was 0-12.5 mM. The detection limit was less than 0.1 mM. The repeatability (%R.S.D.) was 3-5% (n=10). The reproducibility was 5-8% (n=5). Methanol gave no signal at all, but higher alcohols, such as propanol, pentanol and hexanol, gave significant signals, decreasing with increasing length of the carbon chain. The price for one measurement was calculated to be 0.052 euro. The results from measurements with the biosensor were compared to those from an established analysis kit for ethanol. The results correlated well (R(2)=0.9874). The concentration of ethanol in different alcoholic beverages was investigated and correlated well with the concentrations given by the manufacturers. PMID- 16242609 TI - A novel, disposable, screen-printed amperometric biosensor for glucose in serum fabricated using a water-based carbon ink. AB - Screen-printed amperometric glucose biosensors have been fabricated using a water based carbon ink. The enzyme glucose oxidase (GOD) and the electro-catalyst cobalt phthalocyanine were mixed with the carbon ink prior to the screen-printing process; therefore, biosensors are prepared in a one-step fabrication procedure. Optimisation of the biosensor performance was achieved by studying the effects of pH, buffer strength, and applied potential on the analytical response. Calibration studies were performed under optimum conditions, using amperometry in stirred solution, with an operating potential of +500 mV versus SCE. The sensitivity was found to be 1170 nA mM(-1), with a linear range of 0.025-2 mM; the former represents the detection limit. The disposable amperometric biosensor was evaluated by carrying out replicate determinations on a sample of bovine serum. This was achieved by the method of multiple standard additions and included a correction for background currents arising from oxidizable serum components. The mean serum concentration was calculated to be 8.63 mM and compared well with the supplier's value of 8.3 mM; the coefficient of variation was calculated to be 3.3% (n=6). PMID- 16242610 TI - Comparison of surface plasmon resonance spectroscopy and quartz crystal microbalance techniques for studying DNA assembly and hybridization. AB - In this study we evaluate the strengths and weaknesses of surface plasmon resonance (SPR) spectroscopy and quartz crystal microbalance (QCM) technique for studying DNA assembly and hybridization reactions. Specifically, we apply in parallel an SPR instrument and a 5 MHz QCM device with dissipation monitoring (QCM-D) to monitor the assembly of biotinylated DNA (biotin-DNA) on a streptavidin-modified surface and the subsequent target DNA hybridization. Through the parallel measurements, we demonstrate that SPR is more suitable for quantitative analysis of DNA binding amount, which is essential for interfacial DNA probe density control and for the analysis of its effect on hybridization efficiency and kinetics. Although the QCM is not quantitative to the same extent as SPR (QCM measures the total mass of the bound DNA molecules together with the associated water), the dissipation factor of the QCM provides a qualitative measure of the viscoelastic properties of DNA films and the conformation of the bound DNA molecules. The complexity in mass measurement does not impair QCM's potential for a kinetic evaluation of the hybridization processes. For quantification of target DNA, the biotin-DNA modified SPR and QCM sensors are exposed to target DNA with increasing concentration. The plots of SPR/QCM signals versus target DNA concentration show that water entrapment between DNA strands make the QCM sensitivity for the hybridization assay well comparable with that of the SPR, although the intrinsic mass sensitivity of the 5 MHz QCM is approximately 20 times lower. PMID- 16242611 TI - Generation of biochemical response patterns of different substances using a whole cell assay with multiple signaling pathways. AB - Distinctive generation of biochemical response patterns of eight different substances, using an assay based on pigment containing cells, was demonstrated. Xenopus laevis melanophores, transfected with human beta(2)-adrenergic receptor, were seeded in a 96 well microplate and used to generate individual biochemical images through a two transient measuring protocol that contributes to highlight the response signatures of the agents. Adequate signal processing creates distinctive patterns in a time-concentration response space suitable for substance classification. The concept of biochemical images is introduced here. The assays were evaluated both with a standard microplate reader and with a computer screen photo-assisted technique (CSPT) yielding similar results. Since CSPT platforms only demand standard computer sets and web cameras as measuring setup, applications for these kind of assays outside main-laboratories were discussed. PMID- 16242612 TI - Rapid detection of ssDNA and RNA using multi-walled carbon nanotubes modified screen-printed carbon electrode. AB - A method for rapid sensitive detection of DNA or RNA was designed using a composite screen-printed carbon electrode modified with multi-walled carbon nanotubes (MWNTs). MWNTs showed catalytic characteristics for the direct electrochemical oxidation of guanine or adenine residues of signal strand DNA (ssDNA) and adenine residues of RNA, leading to indicator-free detection of ssDNA and RNA concentrations. With an accumulation time of 5 min, the proposed method could be used for detection of calf thymus ssDNA ranging from 17.0 to 345 microg ml(-1) with a detection limit of 2.0 microg ml(-1) at 3 sigma and yeast tRNA ranging from 8.2 microg ml(-1) to 4.1 mg ml(-1). AC impedance was employed to characterize the surface of modified electrodes. The advantages of convenient fabrication, low-cost detection, short analysis time and combination with nanotechnology for increasing the sensitivity made the subject worthy of special emphasis in the research programs and sources of new commercial products. PMID- 16242613 TI - Palm tree peroxidase-based biosensor with unique characteristics for hydrogen peroxide monitoring. AB - Three amperometric enzyme electrodes have been constructed by adsorbing anionic royal palm tree peroxidase (RPTP), anionic sweet potato peroxidase (SPP), or cationic horseradish peroxidase (HRP-C) on spectroscopic graphite electrodes. The resulting H(2)O(2)-sensitive biosensors were characterized both in a flow injection system and in batch mode to evaluate their main bioelectrochemical parameters, such as pH dependency, I(max), K(M)(app), detection limit, linear range, operational and storage stability. The obtained results showed a distinctly different behavior for the plant peroxidase electrodes, demonstrating uniquely superior characteristics of the RPTP-based sensors. The broader linear range observed for the RPTP-based biosensor is explained by a high stability of this enzyme in presence of H(2)O(2). The higher storage and operational stability of RPTP-based biosensor as well as its capability to measure hydrogen peroxide under acidic conditions connect with an extremely high thermal and pH-stability of RPTP. PMID- 16242615 TI - Spore and micro-particle capture on an immunosensor surface in an ultrasound standing wave system. AB - The capture of Bacillus subtilis var. niger spores on an antibody-coated surface can be enhanced when that coated surface acts as an acoustic reflector in a quarter wavelength ultrasonic (3 MHz) standing wave resonator. Immunocapture in such a resonator has been characterised here for both spores and 1 microm diameter biotinylated fluorescent microparticles. A mean spatial acoustic pressure amplitude of 460 kPa and a frequency of 2.82 MHz gave high capture efficiencies. It was shown that capture was critically dependent on reflector thickness. The time dependence of particle deposition on a reflector in a batch system was broadly consistent with a calculated time of 35 s to bring 95% of particles to the coated surface. A suspension flow rate of 0.1 ml/min and a reflector thickness of 1.01 mm gave optimal capture in a 2 min assay. The enhancement of particle detection compared with the control (no ultrasound) situation was x 70. The system detects a total of five particles in 15 fields of view in a 2 min assay when the suspending phase concentration was 10(4) particles/ml. A general expression for the dependence of minimum concentration detectable on; number of fields examined, sample volume flowing through the chamber and assay time shows that, for a practical combination of these variables, the threshold detection concentration can be two orders of magnitude lower. PMID- 16242614 TI - Miniaturized glucose biosensor modified with a nitric oxide-releasing xerogel microarray. AB - An enzyme-based glucose biosensor modified to release nitric oxide (NO) via a xerogel microarray is reported. The biosensor design is as follows: (1) glucose oxidase (GOx) is immobilized in a methyltrimethoxysilane (MTMOS) xerogel layer; (2) a blended polyurethane/hydrophilic polyurethane coating prevents enzyme leaching and imparts selectivity for glucose; and (3) micropatterned xerogel lines (5 microm wide) separated by distances of 5 or 20 microm provide NO-release capability. This configuration allows for increased glucose sensitivity relative to sensors modified with NO-releasing xerogel films since significant portions of the sensor surface remain unmodified. Glucose diffusion to the GOx layer is thus less inhibited. The micropatterned NO-releasing biosensors generate sufficient NO levels to reduce both Pseudomonas aeruginosa and platelet adhesion without significantly compromising the enzymatic activity of GOx. The glucose response, linearity and stability of the NO-releasing micropatterned sensors are reported. PMID- 16242616 TI - Digital image processing-an alternate tool for monitoring of pigment levels in cultured cells with special reference to green alga Haematococcus pluvialis. AB - A method for analyzing carotenoid content in Haematococcus pluvialis, a green alga was developed using digital image processing (DIP) and an artificial neural network (ANN) model. About 90 images of algal cells in various phases of growth were processed with the tools of DIP. A good correlation of R(2)=0.967 was observed between carotenoid content as estimated by analytical method and DIP. Similar correlation was also observed in case of chlorophyll. Since the conventional methods of carotenoid estimation are time consuming and result in loss of pigments during analysis, DIP method was found to be an effective online monitoring method. This method will be useful in measurement of pigments in cultured cells. PMID- 16242617 TI - A multianalyte flow electrochemical cell: application to the simultaneous determination of carbohydrates based on bioelectrocatalytic detection. AB - A multianalyte flow electrochemical cell (MAFEC) for bioanalysis is constructed, characterised and used for simultaneous carbohydrate analysis incorporating mediated amperometric enzyme electrodes. Although multidetection schemes can be addressed with microfabricated systems, it is demonstrated that a "meso" analytical device of low cost can give answers to traditional simultaneous multianalysis problems, being robust, and easy to construct and operate. The cell operates as a radial flow thin-layer device and can achieve mass transport controlled response for fast electrochemical reactions. When appropriate enzymatic electrodes are used the response becomes kinetically limited, but still shows a better than 5% R.S.D. for response to different sugars analysed. All the enzymatic sensors are mediated with different osmium compounds appropriate for each enzyme's mechanism (NAD or PQQ dehydrogenases) in some cases combining multienzyme sensors. All sensors were optimised so that different sugars do not produce interferences to other sensors. Matrix interferences were kept low by operating all sensors at or below 150 mV versus Ag/AgCl. The integrated system was used for the simultaneous detection of fructose, sucrose, glucose, galactose, and lactose, fully characterising the system for these analytes (sensitivity, dynamic range). Cross referenced calibration curves were used for signal treatment and interpretation and it was possible to analyse real juice and milk samples with results agreeing with the standard enzymatic methods for the same analyses with a sampling frequency of more than 100 h(-1). PMID- 16242618 TI - Interface and gate bias dependence responses of sensing organic thin-film transistors. AB - The effects of the exposure of organic thin-film transistors, comprising different organic semiconductors and gate dielectrics, to 1-pentanol are investigated. The transistor sensors exhibited an increase or a decrease of the transient source-drain current in the presence of the analyte, most likely as a result of a trapping or of a doping process of the organic active layer. The occurrence of these two effects, that can also coexist, depend on the gate dielectric/organic semiconductor interface and on the applied gate field. Evidence of a systematic and sizable response enhancement for an OTFT sensor operated in the enhanced mode is also presented. PMID- 16242619 TI - Enhancement of DNA immobilization and hybridization on gold electrode modified by nanogold aggregates. AB - Gold electrodes modified by nanogold aggregates (nanogold electrode) were obtained by the electrodeposition of gold nanoparticles onto planar gold electrode. The Electrochemical response of single-stranded DNA (ssDNA) probe immobilization and hybridization with target DNA was measured by cyclic voltammograms (CV) using methylene blue (MB) as an electroactive indicator. An improving method using long sequence target DNA, which greatly enhanced the response signal during hybridization, was studied. Nanogold electrodes could largely increase the immobilization amount of ssDNA probe. The hybridization amount of target DNA could be increased several times for the manifold nanogold electrodes. The detection limit of nanogold electrode for the complementary 16 mer oligonucleotide (target DNA1) and long sequence 55-mer oligonucleotide (target DNA2) could reach the concentration of 10(-9) mol/L and 10(-11) mol/L, respectively, which are far more sensitive than that of the planar electrode. PMID- 16242620 TI - Cold-plasma modification of oxide surfaces for covalent biomolecule attachment. AB - While many processes have been developed to modify the surface of glass and other oxides for biomolecule attachment, they rely primarily upon wet chemistry and are costly and time-consuming. We describe a process that uses a cold plasma and a subsequent in vacuo vapor-phase reaction to terminate a variety of oxide surfaces with epoxide chemical groups. These epoxide groups can react with amine containing biomolecules, such as proteins and modified oligonucleotides, to form strong covalent linkages between the biomolecules and the treated surface. The use of a plasma activation step followed by an in vacuo vapor-phase reaction allows for the precise control of surface functional groups, rather than the mixture of functionalities normally produced. By maintaining the samples under vacuum throughout the process, adsorption of contaminants is effectively eliminated. This process modifies a range of different oxide surfaces, is fast, consumes a minimal amount of reagents, and produces attachment densities for bound biomolecules that are comparable to or better than commercially available substrates. PMID- 16242621 TI - Novel micromachined silicon sensor for continuous glucose monitoring. AB - The construction and the application properties of a micro-machined silicon sensor for continuous glucose monitoring are presented. The sensor uses the conventional enzymatic conversion of glucose with amperometric detection of H(2)O(2). The innovation is the precise diffusion control of the analyte through a porous silicon membrane into a silicon etched cavity containing the immobilised enzyme. A variation of the number and size of the membrane pores allows to adjust the linear range of the sensor to the respective requirement. The sensor was tested in vitro as well as in clinical studies, being supplied with interstitial fluid. The cavity sensor was designed for a linear range between 0.5 and 20 mM. A signal response time of below 30 s and a signal stability exceeding 1 week is shown. By using a double cavity sensor falsification of the glucose signal by interfering substances can be compensated. In clinical trials the sensor measured continuously in interstitial fluid for up to 18 h without any signal drift and with good correlation to blood glucose reference values. PMID- 16242622 TI - An electrochemical investigation of glucose oxidase at a CdS nanoparticles modified electrode. AB - The direct electrochemistry of glucose oxidase (GOD) adsorbed on a CdS nanoparticles modified pyrolytic graphite electrode was investigated, where the enzyme demonstrated significantly enhanced electron-transfer reactivity. GOD adsorbed on CdS nanoparticles maintained its bioactivity and structure, and could electro-catalyze the reduction of dissolved oxygen, which resulted in a great increase of the reduction peak current. Upon the addition of glucose, the reduction peak current decreased, which could be used for glucose detection. Performance and characteristics of the fabricated glucose biosensor were assessed with respect to detection limit, sensitivity, storage stability and interference exclusion. The results showed that the fabricated biosensor was sensitive and stable in detecting glucose, indicating that CdS nanoparticle was a good candidate material for the immobilization of enzyme in glucose biosensor construction. PMID- 16242623 TI - Design and fabrication of a largely deformable sensorized polymer actuator. AB - Polypyrrole (PPy), with its biomimetic properties such as high power density, large strain, and biocompatibility, is an excellent candidate for a biomimetic microactuator in microrobotics and bioengineering. A polyvinylidene fluorid (PVDF) sensor is also biocompatible, flexible, and chemically stable. Therefore, a PPy actuator is integrated with a PVDF sensor to realize a sensorized polymer actuator. A novel sensorized polymer actuator can accurately measure its bending motion precisely with real time. Experimental results demonstrate the feasibility of the sensorized polymer actuator. The polymer actuator can be actuated while it senses signals induced from the bending motion. In addition, the position of the sensorized polymer actuator can be controlled and adjusted precisely with feedback signals from its embedded sensor at the time of operation. If this system becomes more robust and reliable, its applications are promising and can be realized in cell handling, microrobotics, and microsurgery with the integration of standard microfabrication techniques. PMID- 16242624 TI - Detection of DNA recognition events using multi-well field effect devices. AB - We proposed the multi-well field effect device for detection of charged biomolecules and demonstrated the detection principle for DNA recognition events using quasi-static capacitance-voltage (QSCV) measurement. The multi-well field effect device is based on the electrostatic interaction between molecular charges induced by DNA recognition and surface electrons in silicon through the Si(3)N(4)/SiO(2) thin double-layer. Since DNA molecules and DNA binders such as Hoechst 33258 have intrinsic charges in aqueous solutions, respectively, the charge density changes due to DNA recognition events at the Si(3)N(4) surface were directly translated into electrical signal such as a flat band voltage change in the QSCV measurement. The average flat band shifts were 20.7 mV for hybridization and -13.5 mV for binding of Hoechst 33258. From the results of flat band voltage shifts due to hybridization and binding of Hoechst 33258, the immobilization density of oligonucleotide probes at the Si(3)N(4) surface was estimated to be 10(8) cm(-2). The platform based on the multi-well field effect device is suitable for a simple and arrayed detection system for DNA recognition events. PMID- 16242625 TI - New antibody immobilization method via functional liposome layer for specific protein assays. AB - A specific protein assay system based on functional liposome-modified gold electrodes has been demonstrated. To fabricate such assay system, a liposome layer was initially grown on top of a gold layer. The liposome layer contained two kinds of functional molecules: biotin molecules for the binding sites of streptavidin and N-(10,12-pentacosadiynoic)-acetylferrocene molecules for the facile redox probe in electrochemical detections. Then, streptavidin was attached on the functional liposme-modified layer using the interaction of streptavidin sbiotin complex. On the streptavidin-attached surface, antibody molecules, anti human serum albumin antibodies could be immobilized without any secondary antibodies. AFM imaging of the streptavidin-attached liposome surface revealed a uniform distribution of closely packed streptavidin molecules. In situ quartz crystal microbalance and electrochemical measurements demonstrated that the wanted antibody-antigen reactions should occur with high specificity and selectivity. Our specific antibody assay system, based on a functional liposome modified electrode, can be developed further to yield sophisticated structures for numerous protein chips and immunoassay sensors. PMID- 16242626 TI - Progressive supranuclear palsy: a systematic review. AB - The progressive supranuclear palsy (PSP) is a rapidly progressing degenerative disease belonging to the family of tauophaties, characterized by the involvement of both cortical and subcortical structures. Although the pathogenesis of PSP is still uncertain, genetic, biochemical, and immunohistochemical studies have been performed and are reviewed here. Genetic factors, oxidative damage, neurotoxins, and environmental factors contribute to tau deposition in the cerebral areas involved in PSP. Symptoms originate from the ensuing dysfunction of dopaminergic, GABAergic, cholinergic, and noradrenergic pathways. Recent advances in neuroradiological and instrumental examinations facilitate the diagnosis and have gained new insights into the pathophysiology of PSP, although the primary cause of the disease is unknown and disease-modifying drugs are not yet available. PMID- 16242627 TI - Beta-amyloid accumulation in APP mutant neurons reduces PSD-95 and GluR1 in synapses. AB - Synaptic dysfunction is increasingly viewed as an early manifestation of Alzheimer's disease (AD), but the cellular mechanism by which beta-amyloid (Abeta) may affect synapses remains unclear. Since cultured neurons derived from APP mutant transgenic mice secrete elevated levels of Abeta and parallel the subcellular Abeta accumulation seen in vivo, we asked whether alterations in synapses occur in this setting. We report that cultured Tg2576 APP mutant neurons have selective alterations in pre- and post-synaptic compartments compared to wild-type neurons. Post-synaptic compartments appear fewer in number and smaller, while active pre-synaptic compartments appear fewer in number and enlarged. Among the earliest changes in synaptic composition in APP mutant neurons were reductions in PSD-95, a protein involved in recruiting and anchoring glutamate receptor subunits to the post-synaptic density. In agreement, we observed early reductions in surface expression of glutamate receptor subunit GluR1 in APP mutant neurons. We provide evidence that Abeta is specifically involved in these alterations in synaptic biology, since alterations in PSD-95 and GluR1 are blocked by gamma-secretase inhibition, and since exogenous addition of synthetic Abeta to wild-type neurons parallels changes in synaptic PSD-95 and GluR1 observed in APP mutant neurons. PMID- 16242628 TI - Bilirubin-induced inflammatory response, glutamate release, and cell death in rat cortical astrocytes are enhanced in younger cells. AB - Unconjugated bilirubin (UCB) encephalopathy is a predominantly early life condition resulting from the impairment of several cellular functions in the brain of severely jaundiced infants. However, only few data exist on the age dependent effects of UCB and their association with increased vulnerability of premature newborns, particularly in a sepsis condition. We investigated cell death, glutamate efflux, and inflammatory cytokine dynamics after exposure of astrocytes at different stages of differentiation to clinically relevant concentrations of UCB and/or lipopolysaccharide (LPS). Younger astrocytes were more prone to UCB-induced cell death, glutamate efflux, and inflammatory response than older ones. Furthermore, in immature cells, LPS exacerbated UCB effects, such as cell death by necrosis. These findings provide a basis for the increased susceptibility of premature newborns to UCB deleterious effects, namely when associated with sepsis, and underline how crucial the course of cell maturation can be to UCB encephalopathy during moderate to severe neonatal jaundice. PMID- 16242629 TI - Decreased endocannabinoid levels in the brain and beneficial effects of agents activating cannabinoid and/or vanilloid receptors in a rat model of multiple sclerosis. AB - Recent studies have addressed the changes in endocannabinoid ligands and receptors that occur in multiple sclerosis, as a way to explain the efficacy of cannabinoid compounds to alleviate spasticity, pain, tremor, and other signs of this autoimmune disease. Using Lewis rats with experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis, we recently found a decrease in cannabinoid CB1 receptors mainly circumscribed to the basal ganglia, which could be related to the motor disturbances characteristic of these rats. In the present study, using the same model, we explored the potential changes in several neurotransmitters in the basal ganglia that might be associated with the motor disturbances described in these rats, but we only found a small increase in glutamate contents in the globus pallidus. We also examined whether the motor disturbances and the changes of CB1 receptors found in the basal ganglia of EAE rats disappear after the treatment with rolipram, an inhibitor of type IV phosphodiesterase able to supress EAE in different species. Rolipram attenuated clinical decline, reduced motor inhibition, and normalized CB1 receptor gene expression in the basal ganglia. As a third objective, we examined whether EAE rats also exhibited changes in endocannabinoid levels as shown for CB1 receptors. Anandamide and 2-arachidonoylglycerol levels decreased in motor related regions (striatum, midbrain) but also in other brain regions, although the pattern of changes for each endocannabinoid was different. Finally, we hypothesized that the elevation of the endocannabinoid activity, following inhibition of endocannabinoid uptake, might be beneficial in EAE rats. AM404, arvanil, and OMDM2 were effective to reduce the magnitude of the neurological impairment in EAE rats, whereas VDM11 did not produce any effect. The beneficial effects of AM404 were reversed by blocking TRPV1 receptors with capsazepine, but not by blocking CB1 receptors with SR141716, thus indicating the involvement of endovanilloid mechanisms in these effects. However, a role for CB1 receptors is supported by additional data showing that CP55,940 delayed EAE progression. In summary, our data suggest that reduction of endocannabinoid signaling is associated with the development of EAE in rats. We have also proved that the reduction of CB1 receptors observed in these rats is corrected following treatment with a compound used in EAE such as rolipram. In addition, the direct or indirect activation of vanilloid or cannabinoid receptors may reduce the neurological impairment experienced by EAE rats, although the efficacy of the different compounds examined seems to be determined by their particular pharmacodynamic and pharmacokinetic characteristics. PMID- 16242630 TI - HIV-Tat-mediated Bcl-XL delivery protects retinal ganglion cells during experimental autoimmune optic neuritis. AB - In multiple sclerosis (MS), post-mortem studies of human brain tissue as well as data from animal models have shown that apoptosis of neurons occurs to a significant extent during this disease. As neurodegeneration in MS correlates with permanent neurological deficits in patients, understanding the mechanisms would be an important pre-condition for designing appropriate neuroprotective therapies. Myelin oligodendrocyte glycoprotein-induced experimental autoimmune encephalomyelitis often affects the optic nerve and leads to consecutive apoptosis of retinal ganglion cells (RGCs), the neurons that form its axons. In this study, we fused Bcl-XL to the protein transduction domain of the HIV transactivator of transcription. Thereby, this anti-apoptotic member of the Bcl-2 family was delivered into RGCs of rats with electrophysiologically diagnosed optic neuritis. Transduction of Bcl-XL in our study led to significant rescue of RGCs indicating the relevance of this pathway for neuronal survival under autoimmune inflammatory conditions. PMID- 16242631 TI - Noradrenergic blockade prevents attacks in a model of episodic dysfunction caused by a channelopathy. AB - Episodic neurological dysfunction often results from ion channel gene mutations. Despite knowledge of the mutations, the factors that precipitate attacks in channelopathies are not clear. In humans, mutations of the calcium channel gene CACNA1A are associated with attacks of neurological dysfunction in familial hemiplegic migraine and episodic ataxia type-2. In tottering mice, a mutation in the same gene causes attacks resembling paroxysmal dyskinesia. Stress, a trigger associated with human episodic disorders, reliably elicits attacks in tottering mice. Because noradrenergic neurotransmission is critical to the stress response and because noradrenergic hyperinnervation is observed in tottering mice, the role of norepinephrine in stress-induced attacks was investigated. Drugs that act at alpha-adrenergic receptors to block noradrenergic transmission prevented attacks. However, agents that facilitate noradrenergic neurotransmission failed to induce attacks. These results suggest that, while noradrenergic neurotransmission may be necessary for attacks, an increase in norepinephrine is not sufficient to induce attacks. PMID- 16242632 TI - Cerebrospinal fluid of Alzheimer patients promotes beta-amyloid fibril formation in vitro. AB - Cerebral deposition of amyloid beta-peptide (Abeta) is an invariant feature of Alzheimer's disease (AD). To answer why soluble Abeta does not aggregate to beta amyloid fibrils (fAbeta) in the brain of normal humans, we examined the influence of cerebrospinal fluid (CSF) obtained from AD and non-AD patients on the formation of fAbeta(1-40) and fAbeta(1-42) in vitro, by using fluorescence spectroscopy with thioflavin T and electron microscopy. Although the CSF obtained from both groups inhibited the formation of both fAbeta(1-40) and fAbeta(1-42), the CSF from non-AD patients inhibited the formation of fAbetas more strongly than that from AD patients. In AD patients, the final levels of fAbetas formation showed a significant negative correlation with the Abeta(1-42) level in CSF. These results indicate that fAbeta deposition in the brain of AD may be enhanced by the decrease of specific inhibitory factors and/or by the increase of specific accelerating factors in CSF. PMID- 16242633 TI - Cystatin C modulates neurodegeneration and neurogenesis following status epilepticus in mouse. AB - Brain damaging insults cause alterations in neuronal networks that trigger epileptogenesis, and eventually lead to the appearance of spontaneous seizures. The present experiments were designed to study the cellular expression and functions of a cysteine proteinase inhibitor, cystatin C, whose gene expression is previously shown to be upregulated in the rat hippocampus during status epilepticus (SE)-induced epileptogenesis. The present data showed that the expression of cystatin C protein increased in the mouse hippocampus 7 days following SE and localized mainly to astrocytes and microglia. Acute neuronal death in the hippocampus at 24 h after SE was reduced in cystatin C-/- mice. Also, the basal level of neurogenesis in the subgranular layer of dentate gyrus was decreased in cystatin C-/- mice compared to wildtype littermates. Interestingly, migration of newly born neurons within the granule cell layer was attenuated in cystatin C-/- mice. These data demonstrate that cystatin C has a role in neuronal death and neurogenesis during SE-induced network reorganization. PMID- 16242634 TI - Small non-fibrillar assemblies of amyloid beta-protein bearing the Arctic mutation induce rapid neuritic degeneration. AB - Recent studies suggest that soluble intermediates formed during amyloid beta protein (Abeta) fibrillogenesis are neurotoxic. We studied early aggregation assemblies of wild-type and mutant Abeta bearing the E22G ("Arctic") familial Alzheimer's disease mutation. Using a novel method to present purified, disaggregated Abeta peptides to primary cortical neurons, the detailed temporal pattern of neurotoxicity was assessed. Neurons exposed to Arctic Abeta showed a progressive degeneration that was much more rapid than that with wild-type Abeta, beginning in dendrites and axons and leading to frank cell death. This neurotoxicity paralleled the aggregation process, with neuritic injury first appearing in the presence of small spherical Abeta oligomers, which were followed by a time-dependent elongation of curvilinear Abeta assemblies. One of the earliest neuritic changes was the formation of neurofilament-positive ringlets within axons, which disappeared as neurites followed by cell body degeneration. Our data support the hypothesis that small Abeta intermediates formed early in the aggregation process initiate cellular dysfunction beginning in neurites, leading to neuronal loss. A similar pattern of degeneration may occur during the preclinical and early clinical phases of Alzheimer's disease. PMID- 16242635 TI - Gene expression profiles of reactive astrocytes cultured from dopamine-depleted striatum. AB - We have carried out cDNA array analysis in order to characterize the gene expression profiles of reactive astrocytes from dopamine-depleted striatum. Astrocytes were cultured from the striatum of normal adult rats (adult astrocytes) or adult rats in which the substantia nigra had been lesioned 1 week earlier with 6-hydroxydopamine (reactive astrocytes), an animal model for Parkinson's disease. Three antibodies, 19D1, O1E4, and 13A11, known to label only reactive astrocytes in vivo, stained cultured reactive astrocytes but not adult astrocytes. Analysis with cDNA arrays showed that 38 genes were up-regulated and 75 genes down-regulated in reactive astrocytes compared to normal adult astrocytes. The expression of growth factor and transcription factor genes predominated among the up-regulated genes while those for signal transduction molecules, metabolic enzymes, and receptors for growth factors, hormones, and neurotransmitters predominated among the down-regulated genes. These results will allow the field to address the molecular profiles and functions of astrocytes activated in response to dopamine depletion and may be useful for developing new therapies for Parkinson's disease. PMID- 16242636 TI - Phenotype-dependent functional and pharmacological properties of BK channels in skeletal muscle: effects of microgravity. AB - We investigated the involvement of calcium-activated potassium channel (BK) in skeletal muscle phenotype determination and response to acetazolamide, a BK opener. The BKs of slow-twitching soleus (SOL) and fast-twitching flexor digitorum brevis (FDB) muscles of the rat were investigated by patch-clamp technique. The changes of BK properties following muscle disuse were investigated in the hindlimb-unloaded (HU) rat, an animal model of disuse/microgravity. Two functionally different BKs were found in skeletal muscle. The BK of FDB was sensitive to calcium and to acetazolamide, in contrast the BK of SOL was less sensitive to calcium and was resistant to acetazolamide. After 3-14 days HU, in parallel with the slow-to-fast phenotype transition of the fibers, the BK of SOL acquired properties similar to those of FDB. In skeletal muscle, the BK plays muscle-specific roles contributing to the calcium-dependent phenotype determination/adaptation to disuse. The phenotype specificity of acetazolamide has implications for drug-based therapy of neuromuscular disorders associated to disuse. PMID- 16242637 TI - Locomotor activity and evoked dopamine release are reduced in mice overexpressing A30P-mutated human alpha-synuclein. AB - We have generated a transgenic mouse line overexpressing mutated human A30P alpha synuclein under the control of the prion-related protein promoter. Immunohistology revealed mutated human A30P alpha-synuclein protein in numerous brain areas, but no gross morphological changes, Lewy bodies, or loss of dopaminergic cell bodies. The transgenic mice displayed decreased locomotion, impaired motor coordination, and balance. In vivo voltammetry showed that A30P mice responded to longer stimulation of the ascending dopaminergic pathways with less dopamine release in striatum and had a slower rate of dopamine decline after repeated stimulations or after alpha-methyl-p-tyrosine-HCl treatment. However, dopamine re-uptake or transporter levels were similar in transgenic and control mice. Our data provide evidence that overexpression of mutated human A30P alpha synuclein in mice leads to a reduced size of the dopamine storage pool. This is in agreement with the previously postulated involvement of alpha-synuclein in the turnover of transmitter vesicles and may explain the observed motor deficits in A30P mice. PMID- 16242638 TI - Progressively reduced synaptic vesicle pool size in cultured neurons derived from neuronal ceroid lipofuscinosis-1 knockout mice. AB - The neuronal ceroid lipofuscinoses are a newly-recognized group of lysosomal storage disorders in which neurodegeneration predominates. The pathophysiological basis for this is unknown. In the current paper, we sought to determine whether neurons that lack the enzyme responsible for the infantile form of neuronal ceroid lipofuscinosis (INCL) display abnormalities in culture that could be related to the clinical disorder. Electrophysiological and fluorescent dye studies were performed using cortical neuronal cultures established from postnatal day 2 palmitoyl-protein thioesterase-1 (Ppt1) knockout mice. We found a 30% reduction in synaptic vesicle number per bouton that was progressive with time in culture as well as an elevation in lysosomal pH, whereas a number of passive and active membrane properties of the neurons were normal. The reduction in vesicle pool size was also reflected in a decrease in the frequency of miniature synaptic currents. The progressive and gradual decline in vesicle numbers and miniature event frequency we observed here may be an early indicator of synapse degeneration, in keeping with observations during competitive stimulation at the neuromuscular junction or age-related synapse elimination recently reported by others. PPT1 did not colocalize with synaptic vesicle or synapse markers, suggesting that lysosomal dysfunction leads indirectly to the synaptic abnormalities. We conclude that from an early age, neurons deficient in PPT1 enzyme activity display intrinsically abnormal properties that could potentially explain key features of the clinical disease, such as myoclonus and seizures. PMID- 16242639 TI - Synaptic differences in the patch matrix compartments of subjects with schizophrenia: a postmortem ultrastructural study of the striatum. AB - The striatum processes motor, cognitive, and limbic circuitry. Striatal patch and matrix compartments are organized differently in many aspects including connectivity. Abnormalities in either compartment could have different functional consequences. The present study compares the synaptic organization in the patches and matrix in subjects with schizophrenia (SZ, n = 14) versus normal controls (NC, n = 8). Postmortem striatal tissue was processed for calbindin immunocytochemistry to identify the patch versus matrix compartments, prepared for electron microscopy, and analyzed using stereology. Several synaptic changes were observed in the SZ subjects vs. NCs including a higher density of cortical type synapses in the putamen patch (44% higher) and in the caudate matrix (36% higher) in SZ cases on typical antipsychotic drugs. These changes appeared to be normalized rather than caused by treatment. The abnormal connectivity may represent a failure of normal synaptic pruning and may play a role in limbic or cognitive dysfunction in schizophrenia. PMID- 16242640 TI - Extended period of asymptomatic prion disease after low dose inoculation: assessment of detection methods and implications for infection control. AB - We used quantal dose-titration of a mouse-adapted human transmissible spongiform encephalopathy strain (M470) to compare different analytical methods for their ability to detect asymptomatic brain prion infection after low dose inoculation. At a time point approximately 2.5-fold beyond the mean incubation period of high dose inocula, asymptomatic brain infection was commonly observed using histologic examination, Western blot, and "blind" bioassay following intracerebral inoculation with low titer inocula. At this time point, when a clinical end-point titration would usually be determined, evidence of infection was seen in all healthy animals inoculated with up to 100-fold lower inoculation doses than the lowest causing consistent clinical disease. For the assessment of the presence of asymptomatic infection, we compared different Western immunoblot and histopathological methods in relation to "blind" bioassay using transgenic Tga/20 mice overexpressing mouse prion protein (PrP). Sodium phosphotungstic acid (NaPTA) precipitation of protease-resistant PrP isoforms (PrP(res)) prior to Western blotting was found to approach the sensitivity of the Tga/20 bioassay and was superior to conventional Western blot and histopathological methods, wherein infectivity was commonly found when both of the latter were negative. Re-scaling the original titer by incorporating "blind" transmission data from surviving asymptomatic mice revises the estimate two orders of magnitude higher than the value derived using the conventional clinical disease outcome approach. We also found that the sensitivity of the NaPTA Western blot technique, if used with a diluent such as PBS compared with 10% normal brain homogenate, is adversely affected by up to around 20-fold. We postulate that infectious titer estimates based on more sensitive detection systems such as we report provide a more accurate indication of ultimate transmission risk. PMID- 16242641 TI - Systemic exposure to paraquat and maneb models early Parkinson's disease in young adult rats. AB - In recent years, several lines of evidence have shown an increase in Parkinson's disease (PD) prevalence in rural environments where pesticides are widely used. Paraquat (PQ--herbicide) and maneb (MB--fungicide) are among the compounds suspected to induce neuronal degeneration and motor deficits characteristics of PD. Here, we investigated the effects of PQ and MB on dopaminergic (DA) neuron glia cultures and in vivo in young adult rats. In vitro, PQ led to a loss of DA as compared to non-DA neurons and microglial activation in a dose-dependent manner. Addition of MB had no further effect nor did it lead to microglial activation when used alone. In vivo, 2-month old young adult rats were subjected to intraperitoneal injections of vehicle (n = 4), PQ alone (n = 8), or PQ in combination with MB (n = 8) twice a week for 4 weeks and were sacrificed the day following the last injection. Significant loss of nigral DA neurons was observed in both treatment groups, but a significant decrease in striatal DA fibers was not found. Microglial activation was seen in the nigra of rats subjected to PQ with or without MB. Behavioral analyses demonstrated a mixed pattern of motor impairments, which may have been related to early effects of nigral DA neuronal loss or systemic effects associated with MB exposure in addition to PQ. These results indicate that exposure to PQ with or without MB induces neurodegeneration which might occur via an early inflammatory response in young adult animals. PMID- 16242642 TI - Progressive neurodegeneration in C. elegans model of tauopathy. AB - Discovery of various mutations in the tau gene among frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17) families suggests gain-of-toxic function of wild-type or mutant tau as the mechanism for extensive neuronal loss. We thus generated transgenic nematode (Caenorhabditis elegans) expressing wild type or mutant (P301L and R406W) tau in the touch (mechanosensory) neurons. Whereas the worm expressing wild-type tau showed a small decrease in the touch response across the lifespan, the worm expressing mutant tau displayed a large and progressive decrease. When the touch neurons lost their function, neuritic abnormalities were found prominent, and microtubular loss became remarkable in the later stage. A substantial fraction of degenerating neurons developed tau accumulation in the cell body and neuronal processes. This neuronal dysfunction is not related to the apoptotic process because little recovery from touch abnormality was observed in the ced-3 or ced-4-deficient background. Expression of GSK3 brought about slight deterioration in the touch response, while expression of HSP70 led to some improvement. PMID- 16242644 TI - Constitutive Dyrk1A is abnormally expressed in Alzheimer disease, Down syndrome, Pick disease, and related transgenic models. AB - DYRK1A, dual-specificity tyrosine-regulated kinase 1A, maps to human chromosome 21 within the Down syndrome (DS) critical region. Dyrk1 phosphorylates the human microtubule-associated protein tau at Thr212 in vitro, a residue that is phosphorylated in fetal tau and hyper-phosphorylated in Alzheimer disease (AD) and tauopathies, including Pick disease (PiD). Furthermore, phosphorylation of Thr212 primes tau for phosphorylation by glycogen synthase kinase 3 (GSK-3). The present study examines Dyrk1A in the cerebral cortex of sporadic AD, adult DS with associated AD, and PiD. Increased Dyrk1A immunoreactivity has been found in the cytoplasm and nuclei of scattered neurons of the neocortex, entorhinal cortex, and hippocampus in AD, DS, and PiD. Dyrk1A is found in sarkosyl-insoluble fractions which are enriched in phosphorylated tau in AD brains, thus suggesting a possible association of Dyrk1A with neurofibrillary tangle pathology. Yet, no clear relationship has been observed between tau phosphorylation at Thr212, and GSK-3 and Dyrk1A expression in diseased brains. Transgenic mice bearing a triple tau mutation (G272V, P301L, and R406W) and expressing hyper-phosphoyrylated tau in neurons of the entorhinal cortex, hippocampus, and cerebral neocortex show increased expression of Dyrk1A in individual neurons in the same regions. However, transgenic mice over-expressing Dyrk1A do not show increased phosphorylation of tau at Thr212, thus suggesting that Dyrk1A over-expression does not trigger per se hyper-phosphorylation of tau at Thr212 in vivo. The present observations indicate modifications in the expression of constitutive Dyrk1A in the cytoplasm and nuclei of neurons in various neurodegenerative diseases associated with tau phosphorylation. PMID- 16242643 TI - Minocycline fails to protect cerebellar granular cell cultures against malonate induced cell death. AB - Experimental and clinical studies support the view that the semisynthetic tetracycline minocycline exhibits neuroprotective roles in several models of neurodegenerative diseases, including ischemia, Huntington, Parkinson diseases, and amyotrophic lateral sclerosis. However, recent evidence indicates that minocycline does not always present beneficial actions. For instance, in an in vivo model of Huntington's disease, it fails to afford protection after malonate intrastriatal injection. Moreover, it reverses the neuroprotective effect of creatine in nigrostriatal dopaminergic neurons. This apparent contradiction prompted us to analyze the effect of this antibiotic on malonate-induced cell death. We show that, in rat cerebellar granular cells, the succinate dehydrogenase inhibitor malonate induces cell death in a concentration-dependent manner. By using DFCA, monochlorobimane and 10-N-nonyl-Acridin Orange to measure, respectively, H2O2-derived oxidant species and reduced forms of GSH and cardiolipin, we observed that malonate induced reactive oxygen species (ROS) production to an extent that surpasses the antioxidant defense capacity of the cells, resulting in GSH depletion and cardiolipin oxidation. The pre-treatment for 4 h with minocycline (10-100 microM) did not present cytoprotective actions. Moreover, minocycline failed to block ROS production and to abrogate malonate induced oxidation of GSH and cardiolipin. Additional experiments revealed that minocycline was also unsuccessful to prevent the mitochondrial swelling induced by malonate. Furthermore, malonate did not induce the expression of the iNOS, caspase-3, -8, and -9 genes which have been shown to be up-regulated in several models where minocycline resulted cytoprotective. In addition, malonate-induced down-regulation of the antiapoptotic gene Bcl-2 was not prevented by minocycline, controversially the mechanism previously proposed to explain minocycline protective action. These results suggest that the minocycline protection observed in several neurodegenerative disease models is selective, since it is absent from cultured cerebellar granular cells challenged with malonate. PMID- 16242645 TI - Janus-faced drugs: the double-edged synthetic opiate trade. PMID- 16242646 TI - The kinome is not enough. AB - Mechanism of action studies are essential to link observable effects on cells with molecular targets of small molecules. Caligiuri and coworkers describe how yeast three-hybrid screening identified kinases that might mediate an intriguing tumor cell-specific antiproliferative effect. PMID- 16242648 TI - The stereochemistry of ketoreduction. AB - In this issue of Chemistry & Biology, Leadlay and coworkers report overproduction of a number of ketoreductase domains from modular polyketide synthases. These discrete enzymes allow the stereochemistry of polyketide ketoreduction to be studied in isolation. PMID- 16242647 TI - Small molecules driving myotube fission. AB - In this issue of Chemistry & Biology, Duckmanton et al. have rigorously studied myotube fragmentation, or "cellularization," triggered by microtubule-disrupting agents. They convincingly demonstrate that cellularization remains integral to myogenic dedifferentiation, but is insufficient for reentry of the mononucleate progeny into the cell cycle. PMID- 16242649 TI - Chemical genetics and orphan genetic diseases. AB - Many orphan diseases have been identified that individually affect small numbers of patients but cumulatively affect approximately 6%-10% of the European and United States populations. Human genetics has become increasingly effective at identifying genetic defects underlying such orphan genetic diseases, but little progress has been made toward understanding the causal molecular pathologies and creating targeted therapies. Chemical genetics, positioned at the interface of chemistry and genetics, can be used for elucidation of molecular mechanisms underlying diseases and for drug discovery. This review discusses recent advances in chemical genetics and how small-molecule tools can be used to study and ultimately treat orphan genetic diseases. We focus here on a case study involving spinal muscular atrophy, a pediatric neurodegenerative disease caused by homozygous deletion of the SMN1 (survival of motor neuron 1) gene. PMID- 16242650 TI - Structural basis for the interaction of a vascular endothelial growth factor mimic peptide motif and its corresponding receptors. AB - Vascular endothelial growth factor (VEGF) is central to the survival and development of the vascular and nervous systems. We screened phage display libraries and built a peptide-based ligand-receptor map of binding sites within the VEGF family. We then validated a cyclic peptide, CPQPRPLC, as a VEGF-mimic that binds specifically to neuropilin-1 and VEGF receptor-1. Here, we use NMR spectroscopy to understand the structural basis of the interaction between our mimic peptide and the VEGF receptors. We show that: (1) CPQPRPLC has multiple interactive conformations; (2) receptor binding is mediated by the motif Arg-Pro Leu; and (3) the Pro residue within Arg-Pro-Leu participates in binding to neuropilin-1 but not to VEGF receptor-1, perhaps representing an evolutionary gain-of-function. Therefore, Arg-Pro-Leu is a differential ligand motif to VEGF receptors and a candidate peptidomimetic lead for VEGF pathway modulation. PMID- 16242651 TI - Supramolecular fluorophores for biological studies: phenylene vinylene-amino acid amphiphiles. AB - We report here on a family of self-assembling fluorescent organic amphiphiles with a biomolecular L-lysine hydrophile and a photonically active phenylene vinylene hydrophobe. Unlike conventional amphiphiles, these segmented dendrimers feature a rigid, branched hydrophobe, and have packing characteristics controlled by the ratio of cross-sectional areas of the hydrophobe and hydrophile. In dilute solution, the amphiphiles form supramolecular aggregates, which are easily taken in by cells through an endocytic pathway, and have no discernible effect on cell proliferation or morphology. An analogous pyrene-based amphiphile was cytotoxic, suggesting that cell survival may be linked either to the self-assembling nature of the amphiphiles, or to the specific properties of the phenylene vinylene segment. The combination of photonic and biological components in these amphiphiles provides great potential for applications in sensing or delivery of molecules to intracellular targets. PMID- 16242652 TI - A tailoring activity is responsible for generating polyene amide derivatives in Streptomyces diastaticus var. 108. AB - We recently characterized rimocidin B (3b) and CE-108B (4b) as two polyene amides with improved pharmacological properties, produced by genetically modified Streptomyces diastaticus var. 108. In this work, genetic and biochemical analysis of the producer strain show that the two amides are derived from the parental polyenes rimocidin (3a) and CE-108 (4a) by a post-PKS modification of the free side chain carboxylic acid. This modification is mediated by an amidotransferase activity operating after the biosynthesis of rimocidin (3a) and CE-108 (4a) are completed. Two polyenes, intermediates of the biosynthetic pathway of rimocidin (3a) and CE-108 (4a), were also isolated and shown to have some improved pharmacological properties compared with the final products. PMID- 16242653 TI - A proteome-wide CDK/CRK-specific kinase inhibitor promotes tumor cell death in the absence of cell cycle progression. AB - The identification of molecular determinants of tumor cell survival is an important objective in cancer research. Here, we describe a small-molecule kinase inhibitor (RGB-286147), which, besides inhibiting tumor cell cycle progression, exhibits potent cytotoxic activity toward noncycling tumor cells, but not nontransformed quiescent fibroblasts. Extensive yeast three-hybrid (Y3H)-based proteome/kinome scanning with chemical dimerizers revealed CDK1/2/3/5/7/9 and the less well-characterized CDK-related kinases (CRKs) p42/CCRK, PCTK1/3, and PFTK1 as its predominant targets. Thus, RGB-286147 is a proteome-wide CDK/CRK-specific kinase inhibitor whose further study could yield new insight into molecular determinants of tumor cell survival. Our results also suggest that the [1, 3, 6] tri-substituted-pyrazolo[3,4-d]-pyrimidine-4-one kinase inhibitor scaffold is a promising template for the rational design of kinase inhibitors with potential applications to disease indications other than cancer, such as neurodegeneration, cardiac hypertrophic growth, and AIDS. PMID- 16242654 TI - A single-cell analysis of myogenic dedifferentiation induced by small molecules. AB - An important direction in chemical biology is the derivation of compounds that affect cellular differentiation or its reversal. The fragmentation of multinucleate myofibers into viable mononucleates (called cellularization) occurs during limb regeneration in urodele amphibians, and the isolation of myoseverin, a trisubstituted purine that could apparently activate this pathway of myogenic dedifferentiation in mammalian cells, generated considerable interest. We have explored the mechanism and outcome of cellularization at a single-cell level, and we report findings that significantly extend the previous work with myoseverin. Using a panel of compounds, including a triazine compound with structural similarity and comparable activity to myoseverin, we have identified microtubule disruption as critical for activation of the response. Time-lapse microscopy has enabled us to analyze the fate of identified mononucleate progeny, and directly assess the extent of dedifferentiation. PMID- 16242656 TI - Genes encoding enzymes responsible for biosynthesis of L-lyxose and attachment of eurekanate during avilamycin biosynthesis. AB - The oligosaccharide antibiotic avilamycin A is composed of a polyketide-derived dichloroisoeverninic acid moiety attached to a heptasaccharide chain consisting of six hexoses and one unusual pentose moiety. We describe the generation of mutant strains of the avilamycin producer defective in different sugar biosynthetic genes. Inactivation of two genes (aviD and aviE2) resulted in the breakdown of the avilamycin biosynthesis. In contrast, avilamycin production was not influenced in an aviP mutant. Inactivation of aviGT4 resulted in a mutant that accumulated a novel avilamycin derivative lacking the terminal eurekanate residue. Finally, AviE2 was expressed in Escherichia coli and the gene product was characterized biochemically. AviE2 was shown to convert UDP-D-glucuronic acid to UDP-D-xylose, indicating that the pentose residue of avilamycin A is derived from D-glucose and not from D-ribose. Here we report a UDP-D-glucuronic acid decarboxylase in actinomycetes. PMID- 16242655 TI - Cationic fullerenes are effective and selective antimicrobial photosensitizers. AB - Fullerenes are soccer ball-shaped molecules composed of carbon atoms, and, when derivatized with functional groups, they become soluble and can act as photosensitizers. Antimicrobial photodynamic therapy combines a nontoxic photosensitizer with harmless visible light to generate reactive oxygen species that kill microbial cells. We have compared the antimicrobial activity of six functionalized C(60) compounds with one, two, or three hydrophilic or cationic groups in combination with white light against gram-positive bacteria, gram negative bacteria, and fungi. After a 10 min incubation, the bis- and tris cationic fullerenes were highly active in killing all tested microbes (4-6 logs) under conditions in which mammalian cells were comparatively unharmed. These compounds performed significantly better than a widely used antimicrobial photosensitizer, toluidine blue O. The high selectivity and efficacy exhibited by these photosensitizers encourage further testing for antimicrobial applications. PMID- 16242657 TI - Molecular basis of Celmer's rules: stereochemistry of catalysis by isolated ketoreductase domains from modular polyketide synthases. AB - A system is reported for the recombinant expression of individual ketoreductase (KR) domains from modular polyketide synthases (PKSs) and scrutiny of their intrinsic specificity and stereospecificity toward surrogate diketide substrates. The eryKR(1) and the tylKR(1) domains, derived from the first extension module of the erythromycin PKS and the tylosin PKS, respectively, both catalyzed reduction of (2R, S)-2-methyl-3-oxopentanoic acid N-acetylcysteamine thioester, with complete stereoselectivity and stereospecificity, even though the substrate is not tethered to an acyl carrier protein or an intact PKS multienzyme. In contrast, and to varying degrees, the isolated enzymes eryKR(2), eryKR(5), and eryKR(6) exercised poorer control over substrate selection and the stereochemical course of ketoreduction. These data, together with modeling of diketide binding to KR(1) and KR(2), demonstrate the fine energetic balance between alternative modes of presentation of ketoacylthioester substrates to KR active sites. PMID- 16242658 TI - Evaluation of RNA isolation procedures from human blood and its application for gene expression studies (Sod-1, Sod-2). PMID- 16242659 TI - [Mesenteric panniculitis]. AB - Mesenteric panniculitis also named retractile mesenterite is a rare tumor-like lesion that thickens and shortens the mesentery. It is characterized by the association of inflammation, necrosis or fibrosis involving the adipose tissue of the bowel mesentery. The pathophysiology of this disease remains unclear despite associations with inflammatory diseases or malignancies, especially lymphomas that have been described. When symptomatic, patients may present with abdominal pain, palpable abdominal mass or intestinal obstruction. The disease remains asymptomatic in 30 to 50% of cases. Abdominal CT plays an important role in suggesting the diagnosis and can be useful in distinguishing the several conditions that can mimic mesenteric panniculitis. Nevertheless, pathologic examination of surgical excisional biopsies or sometimes percutaneous biopsies remains necessary to confirm the diagnosis and exclude an underlying infection or malignancy. Medical treatment may consist of therapy with anti inflammatory or immunosuppressive agents and can be proposed in highly symptomatic diseases. Surgical treatment should be exclusively attempted when intestinal obstruction or ischemia occur. Most of the time, it consists in intestinal derivation or segmental resection because complete excision of the lesions is often not possible. Mesenteric panniculitis usually has an uneventful clinical course and resolves spontaneously in a variable delay. PMID- 16242660 TI - [Multivisceral resections of extracolorectal lesions in familial adenomatous polyposis]. AB - AIMS: 1/ To report our experience with multivisceral resections in familial adenomatous polyposis (FAP) for extracolorectal lesions in a cohort of nine patients. 2/ Discuss the long term results of an agressive surgery. PATIENTS AND METHODS: Nine patients (7 males and 2 females) were operated at the University Hospital of Nimes (N=4) and Nantes (N=5). The median age at the first operation was 29 years (range 18-43). A genetic study was performed in six patients and confirmed the mutation on APC gene (exon 11, 13 and 15). All the patients were operated through a classic laparotomy. RESULTS: All patients have underwent a mean of three operations (range 2-5). Eight patients have had initially a total colectomy and 4 underwent subsequent proctectomy. Seven patients had pancreaticoduodenectomy for extensive duodenal adenomas and/or carcinoma. Three had one or multiple small bowel resections for development of carcinoma and one had partial gastric resection for large adenovillous tumor. The median follow up was 25 years (range 15-37) since the first operation. Three patients were died: one of gastric cancer with hepatic metastases, one of peritoneal carcinosis after ileal resection and one of astrocytoma. CONCLUSION: With regard to these nine observations, the authors underline the possibility of multivisceral resection in FAP. Despite a major digestive mutilation, it permits a long survival with acceptable quality of life. The prognosis depends on the aggressiveness of the duodenal or jejunoileal lesions more than of the colorectal tumors if found at the first resection. PMID- 16242661 TI - [Video-assisted and endoscopic parathyroidectomy and thyroidectomy]. PMID- 16242662 TI - [Abrikossoff tumor of the proximal esophagus misdiagnosed as a thyroid nodule]. AB - The diagnosis of thyroid nodules is straightforward and rarely mistaken. We present a case of a paraesophageal granular cell tumor, discovered incidentally during surgery for what it was diagnosed as a suspicious thyroid nodule by ultrasound and FNA. Complete resection was achieved without disruption of the esophageal mucosa. A terminal branch of the recurrent laryngeal nerve had to be resected en bloc with the tumor. Morphological and immunohistochemical diagnosis was established postoperatively. A review of the literature is presented. PMID- 16242663 TI - Commentary on: 'old and new data, new issues: the mitochondrial Deltapsi' by H. Tedeschi. AB - A recent review [H.Tedeschi, Old and New Data, New Issues: the Mitochondrial Deltapsi, Biochim. Biophys. Acta, Bioenerg. (2005)] has questioned the validity of experiments performed more than 30 years ago that provided the first quantitative estimates of the protonmotive force generated by mitochondria. This commentary explains that the review author's confusion stems from ignoring the data in these papers on the trans-membrane pH gradient. PMID- 16242664 TI - Intracellular-free calcium dynamics and F-actin alteration in the formation of macrophage foam cells. AB - The formation of macrophage foam cells, which is the key event in atherosclerosis, occurs by the uptake of oxidized low-density lipoprotein (Ox LDL) via the scavenger receptor (CD36) pathway. Ca(2+) plays an important role in atherosclerosis. However, in the spatiotemporal view, the correlation between kinetic changes of intracellular-free calcium ([Ca(2+)](i)) and the cellular dysfunctions in the formation of macrophage foam cells has not yet been studied in detail. By the use of confocal laser scanning microscope and flow cytometer, we have detected Ca(2+) dynamics, the assembly of F-actin, and the expression of CD36 under the exposure of U937-derived macrophages to Ox-LDL. The uptake of Ox LDL significantly increased [Ca(2+)](i) in U937-derived macrophages in both acute and chronic treatments (P<0.01). In particular, the increases of the induced [Ca(2+)](i) were different in the presence or absence of extracellular Ca(2+) under acute exposure. A time-dependent rise in F-actin assembly and CD36 expression at 12 and 24h was induced, respectively, by Ox-LDL. The spatiotemporal increases of [Ca(2+)](i) induced by Ox-LDL probably have the key effect on the early phrase in the formation of macrophage foam cells. PMID- 16242665 TI - Arsenate respiratory reductase gene (arrA) for Desulfosporosinus sp. strain Y5. AB - Desulfosporosinus sp. strain Y5 is a spore-forming bacterium capable of dissimilatory arsenate reduction coupled to the oxidation of aromatic compounds. In arsenate respiration, the arsenate respiratory reductase (ARR) catalyzes the reduction of arsenate to arsenite. Our objective is to characterize the arrA gene, encoding the ARR, for Desulfosporosinus sp. strain Y5. Oligonucleotide primers were designed based on the few arrA gene sequences available at the time and validated against positive and negative controls. The resulting arrA-amplicon of approximately 2.0kb was cloned and sequenced. The arrA from Desulfosporosinus sp. Y5 is closely related to Desulfitobacterium hafniense (similarity of 77% and 81% at the nucleotide and amino acid levels, respectively). Phylogenetic topology based on the arrA gene was partially congruent with that of 16S rRNA-based analysis. This arrA sequence will support the development of specific tracking probes for Desulfosporosinus sp. Y5 and the molecular characterization and monitoring of dissimilatory arsenate reducing bacteria. PMID- 16242667 TI - A reference map of the Arabidopsis thaliana mature pollen proteome. AB - The male gametophyte (or pollen) plays an obligatory role during sexual reproduction of higher plants. The extremely reduced complexity of this organ renders pollen a valuable experimental system for studying fundamental aspects of plant biology such as cell fate determination, cell-cell interactions, cell polarity, and tip-growth. Here, we present the first reference map of the mature pollen proteome of the dicotyledonous model plant species, Arabidopsis thaliana. Based on two-dimensional gel electrophoresis, matrix-assisted laser desorption/ionization time-of-flight, and electrospray quadrupole time-of-flight mass spectrometry, we reproducibly identified 121 different proteins in 145 individual spots. The presence, subcellular localization, and functional classification of the identified proteins are discussed in relation to the pollen transcriptome and the full protein complement encoded by the nuclear Arabidopsis genome. PMID- 16242668 TI - Estrogen induces rapid decrease in dendritic thorns of CA3 pyramidal neurons in adult male rat hippocampus. AB - Modulation of hippocampal synaptic plasticity by estrogen has been attracting much attention. Thorns of thorny excrescences of CA3 hippocampal neurons are post synaptic regions whose presynaptic partners are mossy fiber terminals. Here we demonstrated the rapid effect of estradiol on the density of thorns of thorny excrescences, by imaging Lucifer Yellow-injected CA3 neurons in adult male rat hippocampal slices. The application of 1nM estradiol induced rapid decrease in the density of thorns on pyramidal neurons within 2h. The estradiol-mediated decrease in the density of thorns was blocked by CNQX (AMPA receptor antagonist) and PD98059 (MAP kinase inhibitor), but not by MK-801 (NMDA receptor antagonist). ERalpha agonist PPT induced the same suppressive effect as that induced by estradiol on the density of thorns, but ERbeta agonist DPN did not affect the density of thorns. Note that a 1nM estradiol treatment did not affect the density of spines in the stratum radiatum and stratum oriens. A search for synaptic ERalpha was performed using purified RC-19 antibody. The localization of ERalpha (67kDa) in the CA3 mossy fiber terminals and thorns was demonstrated using immunogold electron microscopy. These results imply that estradiol drives the signaling pathway including ERalpha and MAP kinase. PMID- 16242669 TI - Association between tubular toxicity of cisplatin and expression of organic cation transporter rOCT2 (Slc22a2) in the rat. AB - Cisplatin is an effective anticancer drug, but has its severe adverse effects, especially nephrotoxicity. The molecular mechanism of cisplatin-induced nephrotoxicity is still not clear. In the present study, we examined the role of rat (r)OCT2, an organic cation transporter predominantly expressed in the kidney, in the tubular toxicity of cisplatin. Using HEK293 cells stably expressing rOCT2 (HEK-rOCT2), we evaluated the cisplatin-induced release of lactate dehydrogenase and the uptake of cisplatin. The release of lactate dehydrogenase and the accumulation of platinum were greater in HEK-rOCT2 cells treated with cisplatin than in mock-transfected cells. Moreover, cimetidine and corticosterone, OCT2 inhibitors, inhibited the cytotoxicity and the transport of cisplatin in HEK rOCT2 cells. Pharmacokinetics of cisplatin was investigated in male and female rats because the renal expression level of rOCT2 was higher in male than female rats. The renal uptake clearance of cisplatin was greater in male than female rats, while the hepatic uptake clearance was similar between the sexes. In addition, glomerular filtration rate and liver function were unchanged, but N acetyl-beta-D-glucosaminidase activity in the bladder urine and the urine volume were markedly increased 2 days after the administration of 2 mg/kg of cisplatin in male rats. Moreover, cisplatin did not induce the elevation of urinary N acetyl-beta-D-glucosaminidase activity in the castrated male rats whose renal rOCT2 level was lower than that of the sham-operated rats. In conclusion, the present results indicated that renal rOCT2 expression was the major determinant of cisplatin-induced tubular toxicity. PMID- 16242670 TI - Role of mechanical and redox stress in activation of mitogen-activated protein kinases in primary cultured rat hepatocytes. AB - Mechanical stress is known to activate signaling cascades, including mitogen activated protein kinase (MAPK) pathways. Although mechanical stress has been implicated in hepatic cirrhosis and liver regeneration following hepatectomy, the signaling pathway(s) that may be activated in hepatocytes in response to mechanical stress have not been determined. Using primary cultured rat hepatocytes to examine cellular signaling in response to mechanical stress associated with medium change, we observed that the phosphorylation status of extracellular signal-regulated kinase 1/2 (ERK1/2), Jun N-terminal kinase and p38 MAPK, but not Akt, was altered. MAPK activation, especially ERK1/2, was rapidly increased within 5 min, followed by a subsequent decrease to below basal levels between 30 min and 1 h following medium change. MAPK/ERK kinase (MEK1/2) phosphorylation followed the same pattern. The phosphorylation of Raf-1 in response to medium change was also consistent with Raf-1 serving as an upstream regulator of MEK1/2-ERK1/2 signaling. Phosphorylation of ERK1/2 was increased by mechanical stress alone, suggesting that mechanical stress may be primarily responsible for ERK1/2 activation in response to medium change. Medium change produced a marked decline in oxidized glutathione and malondialdehyde levels, and the antioxidant N-acetyl-L-cysteine decreased basal ERK1/2 phosphorylation, suggesting a role for oxidative stress in maintaining basal ERK1/2 phosphorylation in cultured hepatocytes. These data suggest that medium change results in immediate activation of the MAPK signaling pathway due to mechanical stress, followed by a subsequent inactivation of MAPK signaling due to a reduction in oxidative stress levels. These processes may be associated with alteration of hepatic hemodynamic circulation observed in hepatic diseases and in liver transplantation. PMID- 16242671 TI - Isolation and characterisation of acanmyotoxin-2 and acanmyotoxin-3, myotoxins from the venom of the death adder Acanthophis sp. Seram. AB - Death adder (genus Acanthophis) venoms display neurotoxic activity but were thought to be devoid of myotoxic components. Studies from our laboratory have shown that some species (i.e. Acanthophis rugosus and Acanthophis sp. Seram) possess venom with myotoxic activity [Wickramaratna JC, Fry BG, Aguilar M, Kini RM, Hodgson WC. Isolation and pharmacological characterisation of a phospholipase A2 myotoxin from the venom of the Irian Jayan death adder (A. rugosus). Br J Pharmacol 2003;138:333-342; Wickramaratna JC, Fry BG, Hodgson WC. Species dependent variations in the in vitro myotoxicity of death adder (Acanthophis) venoms. Toxicol Sci 2003;74:352-360]. The present study describes the isolation and characterisation of two myotoxins (acanmyotoxin-2 and acanmyotoxin-3) from A. sp. Seram venom. Venom was fractionated into approximately 12 major peaks using reverse phase high performance liquid chromatography. Two components caused concentration (0.1-1 microM) dependent inhibition of direct (2 ms, 0.1 Hz, supramaximal V) twitches and an increase in baseline tension in the chick biventer cervicis nerve-muscle. Histological examination of the muscle confirmed damage. PLA2 activity was detected in both acanmyotoxin-2 (390.2+/-19.7 micromol/(min mg); n=4) and acanmyotoxin-3 (14.2+/-7.7 micromol/(min mg); n=4). In comparison, A. sp. Seram whole venom had a specific activity of 461.3+/-90.4 micromol/(min mg) (n=3). Mass spectrometry analysis indicated acanmyotoxin-2 had a mass of 13,082 Da and acanmyotoxin-2 13,896 Da. Acanmyotoxin-2 and acanmyotoxin 3 accounted for approximately 7 and 4% of total venom composition, respectively. N-terminal sequencing of the first 30 amino acids of each toxin indicated they shared some sequence homology with known myotoxins. In conclusion, clinicians should be aware that symptoms of envenoming by some species of death adder may include signs of myotoxicity as well as neurotoxicity. Future studies will investigate the efficacy of the current antivenom treatment against the myotoxic components of A. sp. Seram venom. PMID- 16242673 TI - Hypoxia impedes the formation of chromium DNA-adducts in a cell-free system. AB - The metabolic reduction of hexavalent chromium [Cr(VI)] in the presence of DNA generates several lesions which impede DNA replication and gene transcription. However, the relative contribution of molecular oxygen to Cr-induced genetic damage is unclear. To elucidate the role of dioxygen in Cr genotoxicity, we studied the formation of Cr-induced lesions in DNA treated with either Cr(VI) and the physiological reductant, ascorbic acid (Asc), or Cr(III), under ambient and hypoxic (<1% oxygen) conditions. We found that hypoxia did not impede the reduction of Cr(VI) by Asc throughout a 2 h treatment. In contrast, Cr-DNA binding under these conditions was reduced up to 70% by hypoxia, and a 50-90% decrease in the frequency of Cr-induced Taq polymerase-arresting DNA adducts was also observed. In the presence of Cr(VI)/Asc, formation of Cr-DNA interstrand crosslinks (ICLs) under hypoxia was 50% or less of that under ambient conditions. Kinetic studies found that hypoxia reduced the rate at which Cr interacted with DNA, but not the ultimate steady state level of Cr-DNA binding. The inhibitory effect of hypoxia on Cr(VI)/Asc genotoxicity could not be explained solely by alterations in the reactivity of intermediate Cr(V) species because Cr(III)-DNA binding and Cr(III)-induced ICL formation were also impaired by hypoxia. Moreover, Cr(V) was generated to similar levels in ambient and hypoxic reactions. Hypoxia did not affect ICL formation by the inorganic chemotherapeutic agent cisplatin, suggesting that these effects were specific for Cr(III). Taken together, these results support a role for dioxygen in facilitating the formation of Cr-DNA coordination complexes. PMID- 16242672 TI - Lysophosphatidic acid stimulates cell proliferation in rat chondrocytes. AB - Rat primary chondrocytes express the lysophosphatidic acid (LPA) receptor, LPA1, LPA3, but not LPA2. When chondrocytes were stimulated with LPA, phospholipase C mediated cytosolic calcium increase was dramatically induced. LPA also stimulated two kinds of mitogen-activated protein kinase, extracellular signal-regulated kinase (ERK) and p38 kinase in chondrocytes. In terms of the LPA-mediated functional modulation of chondrocytes, LPA stimulated cellular proliferation. We examined the signaling pathways involved in LPA-mediated cellular proliferation. LPA-induced chondrocyte proliferation was almost completely blocked by 2'-amino 3'-methoxyflavone (PD98059) but not by 4-(4-fluorophenyl)-2-(4 methylsulfinylphenyl)-5-(4-pyridyl)1H-imidazole (SB203580), suggesting that ERK activity is essentially required for the process. Pertussis toxin almost completely inhibited the LPA-induced cellular proliferation and ERK activation, indicating the role of G(i/o) protein(s) in the processes. This study demonstrates the physiological role of LPA on the modulation of rat primary chondrocyte proliferation, and the crucial role played by ERK in the process. PMID- 16242674 TI - Calretinin-containing neurons which co-express parvalbumin and calbindin D-28k in the rat spinal and cranial sensory ganglia; triple immunofluorescence study. AB - The co-expression of calretinin with parvalbumin and calbindin D-28k was examined in the rat cranial and spinal sensory ganglia by triple immunofluorescence method. In the trigeminal and nodose ganglia, 9% and 5% of calretinin immunoreactive neurons, respectively, also contained both parvalbumin- and calbindin D-28k immunoreactivity. These neurons had large cell bodies. In the trigeminal ganglion, they were restricted to the caudal portion. Such neurons were evenly distributed throughout the nodose ganglion. The co-expression could not be detected in the dorsal root, jugular or petrosal ganglia. Nerve fibers which co-expressed all the three calcium-binding proteins were observed in the inferior alveolar nerve but not the infraorbital nerve or palate. In the periodontal ligament, these nerve fibers formed Ruffini-like endings. These findings suggest that (1) the co-expression in trigeminal neurons is intimately related to their peripheral receptive fields; (2) the three calcium-binding proteins (calretinin, parvalbumin, calbindin D-28k) co-expressed in the trigeminal neurons may have mechanoreceptive function in the periodontal ligament. PMID- 16242675 TI - Neuronal zinc stores are modulated by non-steroidal anti-inflammatory drugs: an optical analysis in cultured hippocampal neurons. AB - Zinc chelation and non-steroidal anti-inflammatory drugs (NSAIDs) have been explored as potential neuroprotective agents. However, it remains unknown whether NSAIDs and zinc chelation may converge on a similar cellular process. Using two photon microscopy to observe hippocampal neurons labeled with a zinc-sensitive dye, we provide evidence that three chemically unrelated NSAIDs, niflumic acid, ibuprofen, and naproxen, acutely increase intracellular zinc stores from extracellular metal pools. Phospholipase A2 inhibitors triggered similar responses, suggesting that NSAIDs likely control zinc stores by their activity as cyclooxygenase inhibitors. These results provide evidence for a new link between cyclooxygenase metabolites and the mechanisms controlling neuronal zinc pools. PMID- 16242676 TI - A templating route to nanoporous chitosan materials. AB - Nanoporous chitosan materials in both membrane and bulk form with pore diameter ranges of 10-50 nm were prepared by a novel emulsion-mediated templating method. Considering the biocompatibility and versatility of chitosan, the nanoporous chitosan material is expected to have a wide variety of applications in the biotechnological and biomaterials areas. PMID- 16242677 TI - Unusual conformational behavior of trisaccharides containing N-acetylglucosamine. AB - Protected trisaccharides containing N-acetylglucosamine can adopt unexpected conformations through the formation of hydrogen bonds involving the amide group. This conformational behavior was observed by NMR spectroscopy when three protected trisaccharides were dissolved in deuterated chloroform and to a lesser extent in deuterated dichloromethane. In contrast, NMR spectra of the same analogues acquired in the hydrogen bond-accepting solvents deuterated acetonitrile and dimethylsulfoxide showed that the N-acetylglucosamine residues adopted the expected 4C1 conformation. PMID- 16242678 TI - GLYDE-an expressive XML standard for the representation of glycan structure. AB - The amount of glycomics data being generated is rapidly increasing as a result of improvements in analytical and computational methods. Correlation and analysis of this large, distributed data set requires an extensible and flexible representational standard that is also 'understood' by a wide range of software applications. An XML-based data representation standard that faithfully captures essential structural details of a glycan moiety along with additional information (such as data provenance) to aid the interpretation and usage of glycan data, will facilitate the exchange of glycomics data across the scientific community. To meet this need, we introduce GLYcan Data Exchange (GLYDE) standard as an XML based representation format to enable interoperability and exchange of glycomics data. An online tool () for the conversion of other representations to GLYDE format has been developed. PMID- 16242679 TI - Dexamethasone: effects on neointimal hyperplasia and vessel integrity. PMID- 16242680 TI - Role of ELAV-like RNA-binding proteins HuD and HuR in the post-transcriptional regulation of acetylcholinesterase in neurons and skeletal muscle cells. AB - Over the last few years, several laboratories have focused their attention on elucidating the molecular events that control the expression and localization of acetylcholinesterase (AChE) in neurons and skeletal muscle cells. In this context, results from a number of studies have clearly shown the important contribution of transcriptional events in regulating AChE expression. Specifically, these studies have highlighted the roles of several cis- and trans acting factors that control transcription of the AChE gene in these excitable cells. However, it has also become apparent that changes in the transcriptional activity of the AChE gene cannot fully account for the alterations seen in the overall abundance of AChE transcripts in neurons and muscle cells placed under a variety of experimental conditions. This indicates, therefore, that post transcriptional mechanisms also play a significant role in controlling AChE mRNA expression. With this in mind, we have recently begun to address this issue in greater detail. Here, we provide a summary of our most recent findings dealing with the post-transcriptional regulation of AChE. Together, our studies have shown so far the important contribution of an AU-rich element located in the 3'UTR of AChE transcripts and of the stabilizing RNA-binding proteins of the ELAV like family in regulating AChE expression in differentiating neuronal and muscle cells. PMID- 16242681 TI - Thermal conversions of trimethylsilyl peroxides of linoleic and linolenic acids. AB - The trimethylsilyl (TMS) peroxides/esters of the fatty acid hydroperoxides (9S,10E,12Z)-9-hydroperoxy-10,12-octadecadienoic acid (9-HPOD) and (9Z,11E,13S,15Z)-13-hydroperoxy-9,11,15-octadecatrienoic acid (13-HPOT) were subjected to gas chromatography-mass spectrometry and products formed by thermal rearrangements were identified. The main products were decadienals and the TMS derivatives of 13-oxo-9,11-tridecadienoic acid, epoxyalcohols, hemiacetals, and ketodienes. Oxy radicals as well as epoxyallylic radicals served as intermediates in the formation of these compounds. The thermal TMS peroxide conversions documented provided biomimetic models for enzymatic conversions of fatty acid hydroperoxides and also offered a method to generate an array of oxylipin derivatives of value as reference compounds in GC-MS studies. PMID- 16242682 TI - The role of circulating monocyte chemoattractant protein-1 as a marker of hepatic inflammation in patients with chronic liver disease. AB - We investigated the relationships between plasma monocyte chemoattractant protein 1, serum C-reactive protein, and the degree of hepatic inflammation in patients with chronic liver disease. Monocyte chemoattractant protein-1 concentration was correlated with the histological hepatic inflammation (estimated by the Knodell index) and with standard liver function tests (P<0.01). C-reactive protein was not correlated with any of the variables studied. These results underline the role of monocyte chemoattractant protein-1 in the pathogenesis of liver impairment and suggest that this chemokine may be a reliable marker of inflammation in hepatic derangements. PMID- 16242683 TI - Generation and characterization of Dyt1 DeltaGAG knock-in mouse as a model for early-onset dystonia. AB - A trinucleotide deletion of GAG in the DYT1 gene that encodes torsinA protein is implicated in the neurological movement disorder of Oppenheim's early-onset dystonia. The mutation removes a glutamic acid in the carboxy region of torsinA, a member of the Clp protease/heat shock protein family. The function of torsinA and the role of the mutation in causing dystonia are largely unknown. To gain insight into these unknowns, we made a gene-targeted mouse model of Dyt1 DeltaGAG to mimic the mutation found in DYT1 dystonic patients. The mutated heterozygous mice had deficient performance on the beam-walking test, a measure of fine motor coordination and balance. In addition, they exhibited hyperactivity in the open field test. Mutant mice also showed a gait abnormality of increased overlap. Mice at 3 months of age did not display deficits in beam-walking and gait, while 6 month mutant mice did, indicating an age factor in phenotypic expression as well. While striatal dopamine and 4-dihydroxyphenylacetic acid (DOPAC) levels in Dyt1 DeltaGAG mice were similar to that of wild-type mice, a 27% decrease in 4 hydroxy, 3-methoxyphenacetic acid (homovanillic acid) was detected in mutant mice. Dyt1 DeltaGAG tissues also have ubiquitin- and torsinA-containing aggregates in neurons of the pontine nuclei. A sex difference was noticed in the mutant mice with female mutant mice exhibiting fewer alterations in behavioral, neurochemical, and cellular changes. Our results show that knocking in a Dyt1 DeltaGAG allele in mouse alters their motor behavior and recapitulates the production of protein aggregates that are seen in dystonic patients. Our data further support alterations in the dopaminergic system as a part of dystonia's neuropathology. PMID- 16242685 TI - Synthesis of 3-aryl-5-decapentyl-1,2,4-oxadiazoles possessing antiinflammatory and antitumor properties. AB - A simple, convenient and straightforward synthesis of 3-aryl-1,2,4-oxadiazoles 4a f from arylamidoximes 1a-f and palmitic acid 2 is described. Compounds 4a-f are non-lethal in mice at four times the therapeutic dose (i.p., LD50>1 g kg(-1) of the animals' body weight). These heterocycles have been found to possess antiinflammatory property similar to aspirin and ibuprofen. Three compounds, viz., 4a, d, e have also been evaluated for antitumor activity, where 4d exhibited an excellent activity comparable to lapachol. PMID- 16242686 TI - Disparate release of prolactin and growth hormone from the tilapia pituitary in response to osmotic stimulation. AB - In most teleost fishes, prolactin (PRL) plays a key role in freshwater (FW) adaptation, whereas growth hormone (GH) is involved in seawater (SW) adaptation in salmonids and certain euryhaline species including the tilapia, Oreochromis mossambicus. Consistent with its osmoregulatory activity, PRL release increases in response to physiologically relevant reductions in extracellular osmolality. When dispersed PRL and GH cells from FW-acclimatized fish were incubated in media of varying osmolalities, PRL release increased significantly in response to a 12% reduction in medium osmolality during 1 and 4h of exposure. By contrast, cells from SW-acclimatized fish responded only to a 24% reduction in osmolality. Growth hormone release on the other hand increased whether medium osmolality was reduced or raised. Cell volume increased together with PRL release during the perifusion of dispersed PRL cells in direct proportion to the reduction in medium osmolality. Growth hormone release increased whether GH cell volume increased or decreased. In in vivo studies, circulating PRL levels increased as early as 1h after the transfer of fish from SW to FW, whereas GH levels remained unchanged during 24h of acclimatization. These results indicate that while PRL and GH cells are osmosensitive, the PRL cells respond to reductions in extracellular osmolality in a manner that is consistent with PRL's physiological role in the tilapia. While the rise in GH release following the reduction in osmolality is of uncertain physiological significance, the rise in GH release with the elevation of medium osmolality may be connected to its role in SW adaptation. PMID- 16242687 TI - Cholecystokinin and peptide Y in yellowtail (Seriola quinqueradiata): molecular cloning, real-time quantitative RT-PCR, and response to feeding and fasting. AB - In fish, the peptide hormones cholecystokinin (CCK) and peptide Y (PY) may be involved in pancreatic exocrine secretion, as found with mammalian CCK and peptide YY (PYY); CCK stimulates, whereas PYY inhibits, pancreatic exocrine secretion in mammals. However, there is very little information on these hormones in fish; in particular, the function of PY is still unknown. Therefore, as a first step for understanding the role of CCK and PY in regulating pancreatic exocrine in fish, the cDNAs of CCK and PY were cloned from the digestive tract of yellowtail (Seriola quinqueradiata). The peptide sequence of yellowtail CCK-8, DYLGWMDF, is identical to sequences found in several teleosts. The mature form of yellowtail PY consists of 36 amino acids and has high identity to other fish PYs (88.9-97.2%). Real-time quantitative RT-PCR assays were developed to measure yellowtail CCK and PY mRNA levels. CCK mRNA levels were extremely high in the brain and, among the digestive organs, high concentrations were found in the pyloric caeca and anterior intestine. PY mRNA levels were low in the brain and highest in the anterior intestine. In fasting experiments, mRNA levels of CCK and PY in the anterior intestine showed an antagonistic change after fasting; CCK decreased whereas PY increased. These data suggest that CCK and PY in yellowtail may relate to digestion including, enzyme secretion. PMID- 16242688 TI - Novel effects of CRF on visuomotor behavior and autonomic function in anuran amphibians. AB - Administration of corticotropin-releasing factor (CRF) or exposure to stressors inhibits feeding in anuran amphibians. Since most amphibians rely on visual cues for feeding, these findings have led to the hypothesis that CRF may modulate visuomotor pathways involved in prey detection and predator avoidance. The inhibitory effects of CRF on feeding and prey capture are rapid, and do not appear to require the pituitary-adrenal axis in the short term. CRF neurons are located in key visuomotor processing areas of the anuran brain. Corticotropin releasing factor also has potent stimulatory effects on sympathetic nervous system activity, a key regulatory system involved in both prey capture and predator avoidance. In this review I will discuss the unique model that amphibian species provide for investigating CRF effects on visual perception and visuomotor processing, and will summarize the data suggesting a role for CRF in visuomotor behavior and autonomic function in amphibians. PMID- 16242689 TI - In vitro steroid production by adrenals and kidney-gonads from embryonic southern snow skinks (Niveoscincus microlepidotus): implications for the control of the timing of parturition? AB - In some mammalian species, hormones produced by the embryo(s) at the completion of development are involved in the cascade of events that result in parturition. Our overall aim was to determine whether a similar mechanism exists in viviparous reptiles. The alpine skink Niveoscincus microlepidotus provides a useful model for studies of gestation and parturition in viviparous reptiles as the completion of embryonic development and parturition are temporally separated; ovulation occurs in spring, embryonic development is completed by autumn, but parturition does not occur until the following spring. In this study, we determined how in vitro steroid hormone production by embryonic adrenals (progesterone, corticosterone, and testosterone) and mesonephric kidney-gonads (oestradiol and testosterone) varied during the later stages of gestation. We hypothesised that embryonic adrenals and kidney-gonads were capable of producing steroid hormones in vitro, and that the pattern of production would change as parturition approached, would be influenced by temperature, and would increase in the presence of corticotropin (ACTH) or pregnenolone, respectively. Embryonic adrenals and kidney-gonads were incubated with or without ACTH or the steroid precursor pregnenolone, respectively. Tissues were incubated for 3 h at either 16 or 24 degrees C (preferred body temperature of pregnant N. microlepidotus). Incubation medium was analysed for steroid hormones using radioimmunoassay. Low levels of progesterone were produced in vitro during the later stages of gestation when embryonic adrenals were incubated with ACTH. In vitro corticosterone production by embryonic adrenals also occurred, with greater production occurring when tissues were incubated at 24 degrees C. Testosterone was produced in vitro by both adrenals and kidney-gonads in March (late autumn when embryonic development was complete, but prior to parturition the following spring), with greater production at 16 degrees C. This peak in testosterone production coincided with differentiation and continuing growth of the hemipenes. Low levels of oestradiol were produced in vitro by embryonic kidney-gonads in March. These results provide some support for the hypothesis that corticosterone production by the embryonic adrenal may have a role in determining the timing of parturition in a viviparous reptilian species. Further research is needed to differentiate the activity of various tissues and steroid hormones in control of embryonic development, sexual differentiation, and the potential regulation of gestation and parturition in N. microlepidotus and other viviparous reptiles. PMID- 16242691 TI - Prediction of fixation failure after sliding hip screw fixation. PMID- 16242690 TI - Expression of three proopiomelanocortin subtype genes and mass spectrometric identification of POMC-derived peptides in pars distalis and pars intermedia of barfin flounder pituitary. AB - Proopiomelanocortin (POMC) is a common precursor of adrenocorticotropic hormone (ACTH), melanophore-stimulating hormone (MSH), and endorphin (END). In pituitary gland, POMC receives posttranslational processing by which different peptides are generated in the pars distalis (PD) and pars intermedia (PI). Recently, we cloned three subtypes of the POMC gene in pituitary gland of barfin flounder. The present study was undertaken to elucidate whether the three POMC genes are expressed in both the PD and PI of barfin flounder pituitary, and to identify peptides derived from POMCs in these lobes. We amplified the transcripts of POMC A, -B and -C in both the PD and PI by the reverse transcription-polymerase chain reaction. In situ hybridization also detected signals for these three subtypes in the PD and PI. These results demonstrated that all three POMC genes are expressed in both the PD and PI of barfin flounder pituitary. By mass spectrometric analyses, ACTH-A, Des-acetyl (Ac)-alpha-MSH-A/B (amino acid sequence of alpha-MSH A is identical to that of alpha-MSH-B), beta-MSH-A, corticotropin-like intermediate lobe peptide (CLIP)-A, and N-terminal peptide (N-POMC)-A were identified in the PD. Moreover, Des-Ac-alpha-MSH-A/B, alpha-MSH-A/B, beta-MSH-A and -B, N-beta-lipotropin-A, CLIP-A, N-Ac-beta-END-A(1-41) (C-terminally truncated form of N-Ac-beta-END-A), and N-POMC-A were identified in the PI. Predominant detection of POMC-A-derived peptides indicates the greatest production of POMC-A and no detection of POMC-C-derived peptides indicates the lowest production of POMC-C in both the PD and PI. ACTH-A is specifically produced in the PD, however, the occurrence of Des-Ac-alpha-MSH-A, CLIP-A, and beta-MSH-A shows that the entire POMC-A is further cleaved into small peptides as in the PI. In the PI, some peptides receive modification or truncation as shown by the occurrence of alpha-MSH-A/B and N-Ac-beta-END-A(1-41). These results show differential posttranslational processing of POMC between the PD and PI in barfin flounder pituitary. PMID- 16242692 TI - Entrusting patients with their own plain radiographs increases their availability in fracture clinic. AB - Missing radiographs are a common cause of frustration in many fracture clinics. This can lead to unjustified delays and unnecessary radiation exposure. Patients were entrusted with their own radiographs and asked to bring them back to subsequent clinic consultations. We audited a 14-day-period and compared it with a similar period before implementation of this new protocol. The results showed a significant reduction in the number of missing radiographs by entrusting them with patients. PMID- 16242694 TI - Placental growth factor in the cerebrospinal fluid of women with preeclampsia. AB - OBJECTIVE: Recent data suggest that excess circulating soluble fms-like tyrosine kinase-1 (sFlt-1) may causally relate to preeclampsia. This study investigates the levels of sFlt-1, VEGF, and PlGF in cerebrospinal fluid (CSF) of patients with preeclampsia and normotensive controls. METHODS: CSF was collected from preeclamptic patients (n=15) and controls (n=7) at the time of spinal anesthesia and assayed for PlGF, sFlt-1, and VEGF (total and free) by specific immunoassays. RESULTS: All sought angiogenic factors were measurable. Levels of free PlGF but not sFlt-1 or VEGF (total or free) were increased in CSF of preeclamptic women. There was no significant difference in the ratios of angiogenic factors in the CSF of women with preeclampsia. There was no correlation between levels of angiogenic factors and CSF cell counts or severity of symptoms. CONCLUSION: Elevated levels of PlGF in CSF preeclamptic women may promote vascular permeability and contribute to the hypertensive encephalopathy seen in such patients. PMID- 16242695 TI - Comparison of the efficacy of Burch colposuspension, pubovaginal sling, and tension-free vaginal tape for stress urinary incontinence. AB - OBJECTIVE: To compare the cure rate and confirm the clinical efficacy of the 3 most frequently performed surgical procedures for stress urinary incontinence (SUI). METHODS: Between January 2001 and May 2003, 92 women with SUI were randomly assigned to undergo the Burch colposuspension (n=33), pubovaginal sling (n=28), or tension-free vaginal tape (n=31) at the Department of Obstetrics and Gynecology, Yonsei Medical Center, Seoul, Korea. Patient characteristics, urodynamic study results, cure rates at 3, 6, and 12 months, and complication rates were compared using the chi2 test. RESULTS: There were no statistically significant differences in the cure rates initially, but after 12 months the cure rate of the pubovaginal sling procedure was found to be significantly higher than those of the tension-free vaginal tape or Burch colposuspension procedures. CONCLUSION: The cure rate of the pubovaginal sling procedure was significantly higher after 1 year, but no difference in efficacy was observed between the 2 other procedures. A randomized prospective study of a larger population should be conducted. PMID- 16242696 TI - The association of microalbuminuria with clinical cardiovascular disease and subclinical atherosclerosis in the elderly: the Cardiovascular Health Study. AB - PURPOSE: Microalbuminuria (MA) is a risk factor for cardiovascular disease (CVD). It is not known whether this association is due to the effect of MA on the development of subclinical atherosclerosis or whether MA destabilizes subclinical atherosclerosis, leading to clinical events. METHODS: In a cross-sectional analysis we evaluated 3312 Cardiovascular Health Study participants, age >or=65 years, who had MA testing. Participants were divided into three groups: those without diabetes or hypertension (33%), those with hypertension (52%) and those with diabetes, with or without hypertension (15%). Clinical CVD was defined as presence of coronary heart disease (angina, MI, CABG, PTCA), cerebrovascular disease (stroke, TIA) and peripheral arterial disease (requiring intervention). Among those without clinical disease, subclinical atherosclerosis was defined as increased carotid artery intima-media thickness, decreased ankle arm index or increased left ventricular mass. RESULTS: In each of the three groups of participants, the adjusted odds of prevalent clinical CVD in the presence of MA was 1.70-1.80-fold increased, independent of other risk factors. MA was not associated with risk of subclinical atherosclerosis in those without hypertension or diabetes (OR 1.14 [95% CI 0.59, 2.23]), whereas it was associated with subclinical atherosclerosis in those with hypertension (OR 1.58 [95% CI 1.08, 2.30]) or diabetes (OR 2.51 [95% CI 1.27, 4.94]). CONCLUSION: In the absence of hypertension or diabetes, MA was associated with clinical CVD but not with subclinical atherosclerosis. Thus, a hypothesis may be made that the mechanism of association of MA with clinical vascular disease involves destabilization of the vasculature, leading to clinical disease. PMID- 16242698 TI - Components of the metabolic syndrome and incidence of diabetes in elderly Italians: the Italian Longitudinal Study on Aging. AB - The aim of this study was to explore the relationship among components of the metabolic syndrome and their role in the development of diabetes. We included 2295 subjects, aged 65-84 years, participating in the Italian Longitudinal Study on Aging, a population-based study conducted in 1992 and with a follow-up in 1996. Factor analysis was conducted, separately for diabetic and non-diabetic men and women, using the principle components method and varimax rotations. Factor scores for the baseline were used as independent variables in logistic regressions models to determine risk factors predicting the development of diabetes. Factor analysis among non-diabetic elderly showed two factors for men (body size/insulin resistance, blood pressure/lipids) and three for women (body size, lipids, blood pressure). Among diabetic subjects, three factors emerged for men (body size/lipids/insulin resistance, body size/blood pressure, glucose) and four for women (body size/lipids/insulin resistance, lipids, body size/glucose/insulin resistance, lipids/blood pressure). For non-diabetic men and women, the body size factor (body size/insulin resistance factor for men) was strongly associated with diabetes incidence (OR=2.30, 95% CI 1.41-3.74 and OR=2.06, 95% CI 1.33-3.17, respectively). This study confirms that the metabolic syndrome (MetS) does not recognize one single underlying factor in an elderly cohort and that the obesity factor is a strong predictor of development of new onset diabetes. PMID- 16242697 TI - Difference in atherosclerosis burden in different nations and continents assessed by coronary artery calcium. AB - We utilized coronary artery calcium scores (CACS) to assess differences in atherosclerosis burden between asymptomatic White populations living in continents with different cardiovascular disease rates. The similarities in the genetic pool between Brazilian and Portuguese Caucasian subjects offered an opportunity to assess the influence of environmental factors on the development of atherosclerosis. We reviewed CACS data from 17,563 individuals (12,378 men and 5169 women) collected in the USA (74% of the subjects), Brazil (15% of the subjects) and Portugal (11% of the subjects). CACS was absent in 80 and 88% of Portuguese men and women, compared with 46 and 62% and 33 and 59% of Brazilian and US counterparts (p<0.0001). Although the US subjects showed the lowest prevalence of risk factors they had a higher median (interquartile range) CACS than the Brazilian and the Portuguese cohorts: 4 (0;87), 1 (0;68) and 0 (0;0), respectively (p<0.0001). After adjusting for differences in age and cardiovascular risk factors, US men showed higher relative risk ratios of having any CACS than either Brazilian or Portuguese men. Brazilian and US women did not differ as far as risk of CACS although they demonstrated a greater risk than Portuguese women. In this study, significant differences in CACS were detected among three nations in different continents. The CACS differences paralleled the respective cardiovascular mortality rates. PMID- 16242699 TI - The role of preclinical atherosclerosis in the explanation of educational differences in incidence of coronary events. AB - The associations between educational level, preclinical carotid atherosclerosis and incident coronary events (CE), were investigated in a general population sample of 5399 Swedish middle-aged men and women without history of cardiovascular disease (CVD) over a median follow-up of 8.7 years. Presence of carotid plaque (focal intima-media thickness (IMT) >1.2mm) was determined by B mode ultrasound. In the age- and sex-adjusted model, there was an inverse relationship between educational level and risk of future CE (p for trend=0.002). To explore if there were education differences between groups with similar degrees of preclinical carotid atherosclerosis stratified analyses were made. Those with low educational level without carotid plaque showed a slightly increased hazard rate ratio (HRR), 1.14 (95% CI: 0.65, 1.97), compared to those with high educational level without carotid plaque (reference group). For those with high educational level with carotid plaque the HRR was 1.53 (95% CI: 0.92, 2.55). Having both low educational level and carotid plaque was associated with a HRR of 2.72 (95% CI: 1.72, 4.31). Individuals with plaque generally had more unfavourable cardiovascular risk factor levels, regardless of educational level. However, after risk factor adjustment those with both low education and carotid plaque still had a two-fold increased risk of CE. The results imply that differences in the prevalence of preclinical atherosclerosis seem important in explaining education differences in future coronary morbidity. PMID- 16242700 TI - Elevated HDL is a risk factor for recurrent coronary events in a subgroup of non diabetic postinfarction patients with hypercholesterolemia and inflammation. AB - Recent studies demonstrate important roles for inflammation in development of atherosclerosis with current attention focusing on interactions of inflammation with traditional lipoprotein risk factors. Since the nature of such relationships is largely unknown, we sought to investigate interactions of inflammation with hyperlipidemia in generating cardiovascular risk in a way that would allow recognition of such interactions whether anticipated or not. Thus, we searched for subgroups at high risk for recurrent coronary events in 767 non-diabetic postinfarction patients using an exploratory three-dimensional graphical screening technique with previously established factor analysis-derived inflammatory and lipoprotein-related factors. Results indicated a high-risk patient subgroup defined by factor interaction that was best characterized clinically by high levels of C-reactive protein (CRP) and total cholesterol. Kaplan-Meier and Cox multivariate analysis confirmed high-risk. Additionally, within-subgroup risk related to metabolic, inflammatory, and thrombogenic blood markers was assessed using Cox analysis with results showing only elevated HDL as a significant and independent predictor of risk with hazard ratio, 2.24 (95% CI; 1.12, 4.49; p = 0.023). We conclude that in non-diabetic postinfarction patients, elevated HDL is predictive of risk of recurrent coronary events within a subgroup of patients characterized by simultaneous elevations in serum CRP and total cholesterol. PMID- 16242701 TI - Re "Inexpensive color evaluation of dye-based pressure-sensitive films for plantar studies" T. W. Ng, W.K. Yeong, Journal of Biomechanics 38 (2005) 2130 2133. PMID- 16242702 TI - Lutein in selected Canadian crops and agri-food processing by-products and purification by high-speed counter-current chromatography. AB - This study mainly focused on lutein content in several selected crops grown in southern Ontario, Canada. Marigold flower, a good rotation crop for the control of nematodes in tobacco fields was found to contain 0.77% lutein (after saponification, on dry basis). A high-speed counter-current chromatography (HSCCC) method using a two-phase solvent system consisting of hexane-ethanol water (6:4.5:1.5, v/v/v) was developed for the purification of lutein from the saponification mixture of marigold flower extract. The purity of lutein prepared using this HSCCC method was 97%. Free lutein was found to be the predominant form in three squash varieties, and it was mostly found in the peel rather than the commonly consumed flesh. Sweet Mamma, Buttercup and Pepper squash varieties contained 25.4, 18.4 and 30.1mg/100g fresh weigh (FW) of lutein in the peels, respectively. These concentrations were significantly higher than that in spinach and kale (3.7 and 12.3 mg/100 g FW). beta-Carotene was found most in the peel of Sweet Mamma squash at 13.6 mg/100g FW, whereas it was below 2mg/100g FW in all other samples. Cooking increased extractable free lutein by 22-65% in squash peels. Lutein in Yukon Gold potato was at ca. 0.4 mg/100 g FW. Certain Yukon Gold was also found to contain violaxanthin (0.35 mg/100 g FW). Structures of lutein, beta-carotene and violaxanthin were identified by LC-atmospheric pressure chemical ionization MS in positive ion mode, and by comparing the retention time and UV-vis spectral data with standards. Results from this study suggest the selected crops and agri-food industrial processing by-products of these can be a good source of free lutein. PMID- 16242703 TI - Comparison of comprehensive two-dimensional gas chromatography in conventional and stop-flow modes. AB - A new mode of operation for comprehensive two-dimensional gas chromatography (GC x GC), stop-flow GC x GC, was introduced recently. In this technique, the flow in the primary column is stopped for a brief period of time during each modulation cycle, allowing for a secondary separation time that is longer than would otherwise be permitted by the modulation period in conventional GC x GC. This allows the modulation period and the secondary separation time to become independent variables, and greatly increases the flexibility of the system. In this paper, separations obtained under similar conditions for both conventional and stop-flow GC x GC are presented, demonstrating the advantages that stop-flow GC x GC provides over conventional GC x GC for certain separations. PMID- 16242705 TI - Short-time behavior of mixed diffusion-barrier controlled adsorption. AB - This paper focuses on the short-time adsorption kinetics of nonionic surfactants onto water/air surfaces, analyzed in the context of the mixed diffusion-barrier controlled adsorption modeling framework. Specifically, we reconcile the apparent contradiction between theoretical prediction and experimental observations on the adsorption kinetics mechanism at short times: while the mixed diffusion-barrier controlled model predicts a barrier-controlled adsorption, as well as the impossibility of a diffusion-controlled adsorption at asymptotic short times, the short-time experimental dynamic surface tension (DST) behavior of many nonionic surfactants has been interpreted to result from diffusion-controlled adsorption at asymptotic short times. This is because the short-time experimental DST of these surfactants displays a t variation, which is considered as a fingerprint for the existence of diffusion-controlled adsorption, based on the short-time asymptotic behavior of the diffusion-controlled adsorption model. As a result of this interpretation, the fundamental physical nature of the energy barrier has been proposed to be associated with high surfactant surface concentrations. In this paper, we derive a new nonasymptotic short-time formalism of the mixed diffusion-barrier controlled model to describe surfactant adsorption onto a spherical pendant-bubble surface, including determining the ranges of time and surfactant surface concentration values where the short-time formalism is applicable. Based on this formalism, we find that one can expect to observe an apparent t variation of the DST at short times even for the mixed diffusion barrier controlled adsorption model. We analyze the consequence of this finding by re-evaluating the existing notions of the energy barrier. We conclude that the energy barrier is associated with the adsorption of a single surfactant molecule onto a clean surface. PMID- 16242704 TI - Affinity chromatography matures as bioinformatic and combinatorial tools develop. AB - Affinity chromatography has the reputation of a more expensive and less robust than other types of liquid chromatography. Furthermore, the technique is considered to stand a modest chance of large-scale purification of proteinaceous pharmaceuticals. This perception is changing because of the pressure for quality protein therapeutics, and the realization that higher returns can be expected when ensuring fewer purification steps and increased product recovery. These developments necessitated a rethinking of the protein purification processes and restored the interest for affinity chromatography. This liquid chromatography technique is designed to offer high specificity, being able to safely guide protein manufactures to successfully cope with the aforementioned challenges. Affinity ligands are distinguished into synthetic and biological. These can be generated by rational design or selected from ligand libraries. Synthetic ligands are generated by three methods. The rational method features the functional approach and the structural template approach. The combinatorial method relies on the selection of ligands from a library of synthetic ligands synthesized randomly. The combined method employs both methods, that is, the ligand is selected from an intentionally biased library based on a rationally designed ligand. Biological ligands are selected by employing high-throughput biological techniques, e.g. phage- and ribosome-display for peptide and microprotein ligands, in addition to SELEX for oligonucleotide ligands. Synthetic mimodyes and chimaeric dye-ligands are usually designed by rational approaches and comprise a chloro-triazinlyl scaffold. The latter substituted with various amino acids, carbocyclic, and heterocyclic groups, generates libraries from which synthetic ligands can be selected. A 'lead' compound may help to generating a 'focused' or 'biased' library. This can be designed by various approaches, e.g.: (i) using a natural ligand-protein complex as a template; (ii) applying the principle of complementarity to exposed residues of the protein structure; and (iii) mimicking directly a natural biological recognition interaction. Affinity ligands, based on the peptide structure, can be peptides, peptide-mimetic derivatives (<30 monomers) and microproteins (e.g. 25-200 monomers). Microprotein ligands are selected from biological libraries constructed of variegated protein domains, e.g. minibody, Kunitz, tendamist, cellulose-binding domain, scFv, Cytb562, zinc finger, SpA-analogue (Z-domain). PMID- 16242706 TI - Influence of SDS and two anionic hydrotropes on the micellized state of the triblock copolymer E71G7E71. AB - Complex formation between the triblock copolymer E71G7E71, where E denotes ethylene oxide, the central hydrophobic block, phenyl glycidyl ether, is denoted as G, and the subscripts denote the block length, the surfactant sodium dodecyl sulfate (SDS), and the penicillins cloxacillin and dicloxacillin, which can be considered as hydrotropes has been investigated by dynamic light scattering (DLS) and isothermal titration calorimetry (ITC). The different copolymer/amphiphile systems have been studied at a constant copolymer concentration of 10 g dm(-3) and at different amphiphile concentrations. For all systems, the relaxation time distributions show a well-defined single mode with a shift toward slightly faster times, which indicates that the electrostatic interactions between the complexes is minimum, and so it is possible to estimate a size in terms of the apparent hydrodynamic radii. For ITC the amphiphile solutions were titrated in two different forms: as monomers (below the cmc) and as aggregates. The data indicate that the interaction between the surfactant and the hydrotropes with the copolymer occurs even at the lowest SDS concentration measured. PMID- 16242708 TI - Methods to characterize lymphoid apoptosis in a murine model of autoreactivity. AB - The immune system is shaped by the random generation of lymphocytes followed by apoptosis of self-reactive cells, a process termed negative selection. The survival of these pathogenic cells in the periphery can elicit autoreactivity. We describe the development of a biomarker assay for the detection of pathogenic subpopulations of lymphoid cells in adult non-obese diabetic (NOD) mice based on disease-specific alterations in spontaneous or triggered cell death. Utilizing improved methods of cell separations, two distinct lymphoid cell subpopulations with increased susceptibility to apoptosis were identified and quantified. A subpopulation of CD8+ T cells that constitutes approximately 3-7% of the total CD8+ T cell population underwent apoptosis on exposure to low concentrations of TNF-alpha. Such cells were exclusively detected only in NOD mice with histologic signs of active autoreactivity. The non-T cell compartment of NOD immune system, although resistant to TNF-alpha-induced apoptosis, contained a subpopulation of B cells with spontaneous death by culture alone. The refined detection of small numbers of lymphoid cell subsets with quantifiable differences in apoptosis provides a possible immune biomarker for monitoring disease activity or treatment interventions. PMID- 16242707 TI - A novel capture-ELISA for detection of anti-neutrophil cytoplasmic antibodies (ANCA) based on c-myc peptide recognition in carboxy-terminally tagged recombinant neutrophil serine proteases. AB - Testing for antineutrophil cytoplasmic antibodies (ANCA) reacting with proteinase 3 (PR3) is part of the routine diagnostic evaluation of patients with small vessel vasculitis. For PR3-ANCA detection, capture ELISAs are reported to be superior to direct ELISAs. Standard capture ELISAs, in which PR3 is anchored by anti-PR3 monoclonal antibodies (moAB), have two potential disadvantages. First, the capturing moAB may compete for epitopes recognized by some PR3-ANCA, causing occasional false-negative results. Second, the capture of recombinant PR3 mutant molecules becomes unpredictable as modifications of specific conformational epitopes may not only affect the binding of PR3-ANCA, but also the affinity of the capturing anti-PR3 moAB. Here, we describe a new capture ELISA, and its application for PR3-ANCA detection. This new assay is based on the standardized capture of a variety of different carboxy-terminally c-myc tagged recombinant ANCA target antigens using anti-c-myc coated ELISA plates. Antigen used include c myc tagged human rPR3 variants (mature and pro-form conformations), mouse mature rPR3 and human recombinant neutrophil elastase. This new anti-c-myc-capture ELISA for PR3-ANCA detection has an intra- and inter-assay coefficient of variation of 3.6% to 7.7%, and 15.8% to 18.4%, respectively. The analytical sensitivity and specificity for PR3-ANCA positive serum samples were 93% and 100%, respectively when rPR3 with mature conformation was used as target antigen, and 83% and 100% when the pro-enzyme conformation was employed. In conclusion, this new anti-c-myc capture ELISA compares favorably to our standard capture ELISA for PR3-ANCA detection, enables the unified capture of different ANCA target antigens through binding to a c-myc tag, and allows capture of rPR3 mutants necessary for PR3-ANCA epitope mapping studies. PMID- 16242709 TI - Antioxidant enzymes in Spodoptera littoralis (Boisduval): are they enhanced to protect gut tissues during oxidative stress? AB - The Egyptian armyworm Spodoptera littoralis is a polyphagous insect attacking a number of plant species including those belonging to the Solanaceae and Cruciferaceae families. Its digestive physiology must therefore adapt to the food plant to ensure maximum extraction of nutrients with minimum trade-off in terms of growth retardation by pro-oxidant allelochemicals. To investigate this, the caterpillars of S. littoralis were fed on a semi-artificial diet (Manduca Premix Heliothis Premix) and for 24 h on potato plants (Solanum tuberosum), respectively, at the mature 6th instar, and the levels of oxidative radicals and antioxidant enzymes in their guts were compared. The gut pH, standard redox potential (Eh) and electron availability (pe) revealed that oxidizing conditions prevail which promote oxidation of pro-oxidant allelochemicals in foliage. Oxidative stress in the foregut and midgut tissue and the gut contents was assessed from the generation of superoxide radical, total peroxide content and protein carbonyl content. Antioxidant defense was measured by the activities of superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APOX) and glutathione S-transferase peroxidase (GSTpx). A significant (p < 0.001) increase in the superoxide radical production (in foregut tissue, foregut and midgut contents), concomitant with an increase in total peroxide (in foregut contents) and protein carbonyl levels (in foregut and midgut tissue) were noted in larvae fed on the plants in contrast to those fed the semi-artificial diet. Similarly, a significant up-regulation of antioxidant enzymes SOD (in midgut tissues), CAT (in foregut, midgut tissue and contents), APOX (in foregut contents, midgut tissue and contents) and GSTpx (in foregut tissues) was recorded on the plant diet in comparison to the semi-artificial diet. The pro-oxidant allelochemicals in the plant diet are thus eliminated by the insect at the expense of up-regulation of antioxidative enzymes in response to increased oxidative stress from oxidizable allelochemicals. The results are consistent with the hypothesis that increased concentrations of antioxidants form an important component of the defense of herbivorous insects against both exogenous and endogenous oxidative radicals. PMID- 16242710 TI - The oligomeric distribution of SecYEG is altered by SecA and translocation ligands. AB - The multimeric membrane protein complex translocase mediates the transport of preproteins across and integration of membrane proteins into the inner membrane of Escherichia coli. The translocase consists of the peripheral membrane associated ATPase SecA and the heterotrimeric channel-forming complex consisting of SecY, SecE and SecG. We have investigated the quaternary structure of the SecYEG complex in proteoliposomes. Fluorescence resonance energy transfer demonstrates that SecYEG forms oligomers when embedded in the membrane. Freeze fracture techniques were used to examine the oligomeric composition under non translocating and translocating conditions. Our data show that membrane-embedded SecYEG exists in a concentration-dependent equilibrium between monomers, dimers and tetramers, and that dynamic exchange of subunits between oligomers can occur. Remarkably, the formation of dimers and tetramers in the lipid environment is stimulated significantly by membrane insertion of SecA and by the interaction with translocation ligands SecA, preprotein and ATP, suggesting that the active translocation channel consists of multiple SecYEG complexes. PMID- 16242711 TI - Perturbation of the hierarchical folding of a large RNA by the destabilization of its Scaffold's tertiary structure. AB - The P4-P6 domain serves as a scaffold against which the periphery and catalytic core organize and fold during Mg2+-mediated folding of the Tetrahymena thermophila ribozyme. The most prominent structural motif of the P4-P6 domain is the tetraloop-tetraloop receptor interaction which "clamps" the distal parts of its hairpin-like structure. Destabilization of the tertiary structure of the P4 P6 domain by perturbation of the tetraloop-tetraloop receptor interaction alters the Mg2+-mediated folding pathway. The folding hierarchy of P5c approximately P4 P6 > periphery > catalytic core that is a striking attribute of the folding of the wild-type RNA is abolished. The initial steps in folding of the mutant RNA are > or =50-fold faster than those of the wild-type ribozyme with the earliest observed tertiary contacts forming around regions known to specifically bind Mg2+. The interaction between the mutant tetraloop and the tetraloop receptor appears coincidently with slowly forming catalytic core tertiary contacts. Thus, the stability conferred upon the P4-P6 domain by the tetraloop-tetraloop receptor interaction dictates the preferred folding pathway by stabilizing an early intermediate. A sub-denaturing concentration of urea diminishes the early barrier to folding the wild-type ribozyme along with complex effects on the subsequent steps of folding the wild-type and mutant RNA. PMID- 16242712 TI - High-resolution crystal structure of AKR11C1 from Bacillus halodurans: an NADPH dependent 4-hydroxy-2,3-trans-nonenal reductase. AB - Aldo-keto reductase AKR11C1 from Bacillus halodurans, a new member of aldo-keto reductase (AKR) family 11, has been characterized structurally and biochemically. The structures of the apo and NADPH bound form of AKR11C1 have been solved to 1.25 A and 1.3 A resolution, respectively. AKR11C1 possesses a novel non-aromatic stacking interaction of an arginine residue with the cofactor, which may favor release of the oxidized cofactor. Our biochemical studies have revealed an NADPH dependent activity of AKR11C1 with 4-hydroxy-2,3-trans-nonenal (HNE). HNE is a cytotoxic lipid peroxidation product, and detoxification in alkaliphilic bacteria, such as B.halodurans, plays a crucial role in survival. AKR11C1 could thus be part of the detoxification system, which ensures the well being of the microorganism. The very poor activity of AKR11C1 on standard, small substrates such as benzaldehyde or DL-glyeraldehyde is consistent with the observed, very open active site lacking a binding pocket for these substrates. In contrast, modeling of HNE with its aldehyde function suitably positioned in the active site suggests that its elongated hydrophobic tail occupies a groove defined by hydrophobic side-chains. Multiple sequence alignment of AKR11C1 with the highly homologous iolS and YqkF proteins shows a high level of conservation in this putative substrate-binding site. We suggest that AKR11C1 is the first structurally characterized member of a new class of AKRs with specificity for substrates with long aliphatic tails. PMID- 16242713 TI - Cell-free selection of zinc finger DNA-binding proteins using in vitro compartmentalization. AB - We have exploited emulsion-based in vitro compartmentalization (IVC) to devise a method for the selection of zinc finger proteins (ZFPs) on the basis of their DNA binding specificity. A library of ZFPs fused to a C-terminal peptide tag is encoded by a set of DNA cassettes that are prepared wholly in vitro. In addition to the ZFP gene, each DNA cassette also carries a given DNA target binding site sequence for which one wishes to isolate ZFP binders. An aliquot of the library is added to bacterial S30 extract and emulsified in mineral oil so that most of the aqueous droplets contain, on average, no more than one gene. If an intra compartmentally expressed ZFP binds specifically to its encoding DNA via the target binding site, the complex can be purified by affinity capture via the peptide tag after breaking the emulsion, thus rescuing the gene. We present proof of-principle for this IVC selection method by selecting a specific high-affinity ZFP gene from a high background of a related gene. We also propose that high affinity ZFPs can be used as genotype-phenotype linkages to enable selection of other proteins using IVC. PMID- 16242714 TI - Reversed DNA strand cleavage specificity in initiation of Cre-LoxP recombination induced by the His289Ala active-site substitution. AB - During the first steps of site-specific recombination, Cre protein cleaves and religates a specific homologous pair of LoxP strands to form a Holliday junction (HJ) intermediate. The HJ is resolved into recombination products through exchange of the second homologous strand pair. CreH289A, containing a His to Ala substitution in the conserved R-H-R catalytic motif, has a 150-fold reduced recombination rate and accumulates HJs. However, to produce these HJs, CreH289A exchanges the opposite set of strands compared to wild-type Cre (CreWT). To investigate how CreH289A and CreWT impose strand exchange order, we characterized their reactivities and strand cleavage preferences toward LoxP duplex and HJ substrates containing 8bp spacer substitutions. Remarkably, CreH289A had different and often opposite strand exchange preferences compared to CreWT with nearly all substrates. CreH289N was much less perturbed, implying that overall recombination rate and strand exchange depend more on His289 hydrogen bonding capability than on its acid/base properties. LoxP substitutions immediately 5' (S1 nucleotide) or 3' (S1' nucleotide) of the scissile phosphate had large effects on substrate utilization and strand exchange order. S1' substitutions, designed to alter base-unstacking events concomitant with Cre-induced LoxP bending, caused HJ accumulation and dramatically inverted the cleavage preferences. That pre-formed HJs were resolved via either strand in vitro suggests that inhibition of the "conformational switch" isomerization required to trigger the second strand exchange accounts for the observed HJ accumulation. Rather than reflecting CreWT behavior, CreH289A accumulates HJs of opposite polarity through a combination of its unique cleavage specificity and an HJ isomerization defect. The overall implication is that cleavage specificity is mediated by sequence-dependent DNA deformations that influence the scissile phosphate positioning and reactivity. A role of His289 may be to selectively stabilize the "activated" phosphate conformation in order to promote cleavage. PMID- 16242715 TI - Substrate access to the active sites in aminopeptidase T, a representative of a new metallopeptidase clan. AB - Aminopeptidase T (AmpT) from Thermus thermophilus is a metalloexopeptidase with no similarity to prototypical metallopeptidases with an HExxH or HxxEH motif. The crystal structure of the Staphylococcus aureus homologue of AmpT, which is known as aminopeptidase S (AmpS), has been reported recently. This structure revealed a dimeric protein with a very unusual, elongated shape and a large internal cavity. The active sites were found on the inner walls of the cavity and were entirely shielded from the environment, which suggested either that the dimer in the crystals was not physiologically relevant, or that an inactive conformation had been crystallized. Here, we show by gel-filtration and analytical ultracentrifugation that AmpT, like AmpS, forms dimers in solution, and we present the structure of AmpT in a crystal form with five protomers in the asymmetric unit. The five protomers take conformations that range from fully closed, as in the AmpS structure, to nearly open, so that the active site is almost directly accessible. The different conformations indicate flexibility between the AmpT N and C-domains, and explain how AmpT can be active, although the unusual AmpS dimerization mode applies to AmpT as well. PMID- 16242716 TI - Crystal structures of an NAD kinase from Archaeoglobus fulgidus in complex with ATP, NAD, or NADP. AB - NAD kinase is a ubiquitous enzyme that catalyzes the phosphorylation of NAD to NADP using ATP or inorganic polyphosphate (poly(P)) as phosphate donor, and is regarded as the only enzyme responsible for the synthesis of NADP. We present here the crystal structures of an NAD kinase from the archaeal organism Archaeoglobus fulgidus in complex with its phosphate donor ATP at 1.7 A resolution, with its substrate NAD at 3.05 A resolution, and with the product NADP in two different crystal forms at 2.45 A and 2.0 A resolution, respectively. In the ATP bound structure, the AMP portion of the ATP molecule is found to use the same binding site as the nicotinamide ribose portion of NAD/NADP in the NAD/NADP bound structures. A magnesium ion is found to be coordinated to the phosphate tail of ATP as well as to a pyrophosphate group. The conserved GGDG loop forms hydrogen bonds with the pyrophosphate group in the ATP-bound structure and the 2' phosphate group of the NADP in the NADP-bound structures. A possible phosphate transfer mechanism is proposed on the basis of the structures presented. PMID- 16242717 TI - Crystal structure of the complex between the monomeric form of Toxoplasma gondii surface antigen 1 (SAG1) and a monoclonal antibody that mimics the human immune response. AB - Toxoplasma gondii, the intracellular parasite responsible for toxoplasmosis infects more than one-third of the world population and can be life-threatening for fetuses and immunocompromised patients. The surface protein SAG1 is an important immune target, which provides a strong immune response against the invasive tachyzoite while the other forms of the parasite, devoid of SAG1 at their surface, are multiplying. In addition to this role as a "hot spot" decoy, SAG1 is predicted to act as an adhesin during host-cell attachment through its binding to proteoglycans. To begin to understand the relationships between SAG1 epitopes and the ligand-binding site, we have solved the crystal structure of the monomeric form of T.gondii SAG1 complexed to a Fab derived from a monoclonal antibody raised against tachyzoite particles. This antibody competes strongly with human Toxoplasma-specific sera, suggesting that its epitope is part of an immunodominant region present on the surface of SAG1. The structure reveals that this conformational epitope, located within the SAG1 N-terminal domain, does not overlap with the proposed ligand-binding pocket. This study provides the first structural description of the monomeric form of SAG1, and significant insights into its dual role of adhesin and immune target during parasite infection. PMID- 16242718 TI - Electrostatics of the intracellular vestibule of K+ channels. AB - Previous calculations using continuum electrostatic calculations showed that a fully hydrated monovalent cation is electrostatically stabilized at the center of the cavity of the KcsA potassium channel. Further analysis demonstrated that this cavity stabilization was controlled by a balance between the unfavorable reaction field due to the finite size of the cavity and the favorable electrostatic field arising from the pore helices. In the present study, continuum electrostatic calculations are used to investigate how the stability of an ion in the intracellular vestibular cavity common to known potassium channels is affected as the inner channel gate opens and the cavity becomes larger and contiguous with the intracellular solution. The X-ray structure of the calcium-activated potassium channel MthK, which was crystallized in the open state, is used to construct models of the KcsA channel in the open state. It is found that, as the channel opens, the barrier at the helix bundle crossing decreases to approximately 0 kcal/mol, but that the ion in the cavity is also significantly destabilized. The results are compared and contrasted with additional calculations performed on the KvAP (voltage-activated) and KirBac1.1 (inward rectifier) channels, as well as models of the pore domain of Shaker in the open and closed state. In conclusion, electrostatic factors give rise to energetic constraints on ion permeation that have important functional consequences on the various K+ channels, and partly explain the presence or absence of charged residues near the inner vestibular entry. PMID- 16242719 TI - A single disulfide bond differentiates aggregation pathways of beta2 microglobulin. AB - Deposition of wild-type beta2-microglobulin (beta2m) into amyloid fibrils is a complication in patients undergoing long-term hemodialysis. The native beta sandwich fold of beta2m has a highly conserved disulfide bond linking Cys25 and Cys80. Oxidized beta2m forms needle-like amyloid fibrils at pH 2.5 in vitro, whereas reduced beta2m, at acid pH, in which the intra-chain disulfide bond is disrupted, cannot form typical fibrils. Instead, reduced beta2m forms thinner and more flexible filaments. To uncover the difference in molecular mechanisms underlying the aggregation of the oxidized and reduced beta2m, we performed molecular dynamics simulations of beta2m oligomerization under oxidized and reduced conditions. We show that, consistent with experimental observations, the oxidized beta2m forms domain-swapped dimer, in which the two proteins exchange their N-terminal segments complementing each other. In contrast, both dimers and trimers, formed by reduced beta2m, are comprised of parallel beta-sheets between monomers and stabilized by the hydrogen bond network along the backbone. The oligomerized monomers are in extended conformations, capable of further aggregation. We find that both reduced and oxidized dimers are thermodynamically less stable than their corresponding monomers, indicating that beta2m oligomerization is not accompanied by the formation of a thermodynamically stable dimer. Our studies suggest that the different aggregation pathways of oxidized and reduced beta2m are dictated by the formation of distinct precursor oligomeric species that are modulated by Cys25-Cys80 disulfide-bonds. We propose that the propagation of domain swapping is the aggregation mechanism for the oxidized beta2m, while "parallel stacking" of partially unfolded beta2m is the aggregation mechanism for the reduced beta2m. PMID- 16242720 TI - A comparison of antiangiogenic therapies for the prevention of liver metastases. AB - Angiogenesis is essential for solid tumor growth. Although successful antiangiogenic therapies have been demonstrated in animal models, a systematic comparison of the efficacy of different antiangiogenic factors has not been described in the hepatic environment. To address this issue, CT26 murine colon carcinoma cells were transfected with retroviral vectors encoding murine endostatin (mEndostatin), human angiostatin (hAngiostatin), murine-soluble vascular endothelial growth factor receptor-2, (msFlk-1), or murine-soluble Tie2 (msTie2). The transfected cells were then subjected to another round of transfection with a luciferase cDNA-encoding retroviral vector. Expression of these putative antiangiogenic proteins inhibited the proliferation of human umbilical vein endothelial cells in vitro but not tumor cells. To examine effects on tumor growth in vivo, modified cells were delivered via intrasplenic injection into BALB/c mice to induce liver metastases. Tumor burden was measured weekly by bioluminescence. Growth of hepatic metastases in vivo was significantly reduced in mice that were administered cells expressing msTie2 (76% reduction compared with control cells 21 days after intrasplenic inoculation; P < 0.05). Similar results were observed with cells that expressed msFlk-1 and hAngiostatin. However, expression of mEndostatin had no significant effect on the growth of liver metastases compared with control animals. These findings indicate that multiple antiangiogenic pathways are necessary for the growth of hepatic metastases, and each of these pathways is a potential clinically relevant antiangiogenic target for the treatment of this disease. PMID- 16242721 TI - Wound healing potential of Cassia fistula on infected albino rat model. AB - BACKGROUND: Infection is the major problem to treat the wound. Antibiotic resistance by the pathogenic microorganism renders drug ineffective and calls for improved designing and development of new drugs. New approach has been developed to isolate active components from botanicals. Our aim was to investigate the potential of Cassia fistula to treat the infected wound on albino rat model. MATERIAL AND METHODS: The alcohol extract of C. fistula leaves was analyzed for antibacterial effect against Staphylococcus aureus ATCC 29213 and Pseudomonas aeruginosa ATCC 27853. Formulated ointment was topically applied on the infected wound. Wound reduction rate, histological analysis, biochemical analysis, and gelatin zymography were obtained to assess the healing pattern. RESULTS: C. fistula treated rats showed, better wound closure, improved tissue regeneration at the wound site, and supporting histopathological parameters pertaining to wound healing. Biochemical analysis and matrix metalloproteinases expression correlated well with the results thus confirming efficacy of C. fistula in the treatment of the infected wound. CONCLUSION: Along with the other activities such as antitumor, antioxidant, hypoglycemic, hepatoprotective, antibacterial, hypocholesterolaemic, and antidiabetic activity, the healing potential of C. fistula provides a scientific rationale for the traditional use of this plant in the management of infected dermal wound and can be further investigated as a substitute to treat infected wounds without using synthetic antibiotics. PMID- 16242723 TI - Investigating the presence of human herpesvirus 7 and 8 in multiple sclerosis and normal control brain tissue. AB - Multiple sclerosis (MS) is an important demyelinating disease of the central nervous system, the aetiology of which is thought to have a possible viral component. In this study we investigated the possible involvement in MS of two herpes viruses: the neurotropic human herpesvirus 7 (HHV-7) and the related human herpesvirus 8 (HHV-8). Utilising fluorescent in situ hybridisation (FISH) techniques, we examined human post mortem tissues for the presence of immediate early and late viral gene or protein expression in MS patient normal appearing white matter (NAWM), lesional tissue and normal control brain samples. HHV-7 and/or HHV-8 mRNA or protein was detected in some individuals in all three sample categories and was restricted to oligodendrocytes, as determined by double mRNA FISH analysis or immuno fluorescence (IF). No samples showed evidence of viral mRNA when subjected to RT-PCR on extracted ribonucleic acid. We therefore conclude that there is little evidence in our particular sample cohort to suggest involvement of either HHV-7 or HHV-8 in MS. PMID- 16242724 TI - Ecotypes of the Mycobacterium tuberculosis complex. AB - A phylogeny of the Mycobacterium tuberculosis complex has recently shown that the animal-adapted strains are found in a single lineage marked by the deletion of chromosomal region 9 (RD9) [Brosch et al., 2002. A new evolutionary scenario for the Mycobacterium tuberculosis complex. Proc. Natl Acad. Sci. USA 99 (6), 3684 3689]. We have obtained the spoligotype patterns of the RD9 deleted strains used to generate this new evolutionary scenario and we show that the presence of spoligotype spacers 3, 9, 16, 39, and 40-43 is phylogenetically informative in this lineage. We have used the phylogenetically informative spoligotype spacers to screen a database of spoligotype patterns and have identified further members of a group of strains apparently host-adapted to antelopes. The presence of the spoligotype spacers is congruent with the phylogeny generated by chromosomal deletions, suggesting that recombination is rare or absent between strains of this lineage. The phylogenetically informative spacers, in concert with the previously identified single nucleotide mutations and chromosomal deletions, can be used to identify a series of clades in the RD9 deleted lineage each with a separate host preference. Finally, we discuss the application of the ecotype concept to this series of clades and suggest that the M. tuberculosis complex may best be described as a series of host-adapted ecotypes. PMID- 16242725 TI - Role of selection in fixation of gene duplications. AB - New genes commonly appear through complete or partial duplications of pre existing genes. Duplications of long DNA segments are constantly produced by rare mutations, may become fixed in a population by selection or random drift, and are subject to divergent evolution of the paralogous sequences after fixation, although gene conversion can impede this process. New data shed some light on each of these processes. Mutations which involve duplications can occur through at least two different mechanisms, backward strand slippage during DNA replication and unequal crossing-over. The background rate of duplication of a complete gene in humans is 10(-9)-10(-10) per generation, although many genes located within hot-spots of large-scale mutation are duplicated much more often. Many gene duplications affect fitness strongly, and are responsible, through gene dosage effects, for a number of genetic diseases. However, high levels of intrapopulation polymorphism caused by presence or absence of long, gene containing DNA segments imply that some duplications are not under strong selection. The polymorphism to fixation ratios appear to be approximately the same for gene duplications and for presumably selectively neutral nucleotide substitutions, which, according to the McDonald-Kreitman test, is consistent with selective neutrality of duplications. However, this pattern can also be due to negative selection against most of segregating duplications and positive selection for at least some duplications which become fixed. Patterns in post fixation evolution of duplicated genes do not easily reveal the causes of fixations. Many gene duplications which became fixed recently in a variety of organisms were positively selected because the increased expression of the corresponding genes was beneficial. The effects of gene dosage provide a unified framework for studying all phases of the life history of a gene duplication. Application of well-known methods of evolutionary genetics to accumulating data on new, polymorphic, and fixed duplication will enhance our understanding of the role of natural selection in the evolution by gene duplication. PMID- 16242726 TI - The effect of genetic drift on the variance/covariance components generated by multilocus additive x additive epistatic systems. AB - The effect of population bottlenecks on the components of the genetic variance/covariance generated by n neutral independent additive x additive loci has been studied theoretically. In its simplest version, this situation can be modelled by specifying the allele frequencies and homozygous effects at each locus, and an additional factor measuring the strength of the n-th order epistatic interaction. The variance/covariance components in an infinitely large panmictic population (ancestral components) were compared with their expected values at equilibrium over replicates randomly derived from the base population, after t bottlenecks of size N (derived components). Formulae were obtained giving the derived components (and the between-line variance) as functions of the ancestral ones (alternatively, in terms of allele frequencies and effects) and the corresponding inbreeding coefficient F(t). The n-th order derived component of the genetic variance/covariance is continuously eroded by inbreeding, but the remaining components may increase initially until a critical F(t) value is attained, which is inversely related to the order of the pertinent component, and subsequently decline to zero. These changes can be assigned to the between-line variances/covariances of gene substitution and epistatic effects induced by drift. Numerical examples indicate that: (1) the derived additive variance/covariance component will generally exceed its ancestral value unless epistasis is weak; (2) the derived epistatic variance/covariance components will generally exceed their ancestral values unless allele frequencies are extreme; (3) for systems showing equal ancestral additive and total non-additive variance/covariance components, those including a smaller number of epistatic loci may generate a larger excess in additive variance/covariance after bottlenecks than others involving a larger number of loci, provided that F(t) is low. Our results indicate that it is unlikely that the rate of evolution may be significantly accelerated after population bottlenecks, in spite of occasional increments of the derived additive variance over its ancestral value. PMID- 16242727 TI - The Maynard Smith model of sympatric speciation. AB - The paper entitled "Sympatric speciation," which was published by John Maynard Smith in 1966, initiated the development of mathematical models aiming to identify the conditions for sympatric speciation. A part of that paper was devoted to a specific two-locus, two-allele model of sympatric speciation in a population occupying a two-niche system. Maynard Smith provided some initial numerical results on this model. Later, Dickinson and Antonovics (1973) and Caisse and Antonovics (1978) performed more extensive numerical studies on the model. Here, I report analytical results on the haploid version of the Maynard Smith model. I show how the conditions for sympatric and parapatric speciation and the levels of resulting genetic divergence and reproductive isolation are affected by the strength of disruptive selection and nonrandom mating, recombination rate, and the rates of male and female dispersal between the niches. PMID- 16242728 TI - Synergy and discounting of cooperation in social dilemmas. AB - The emergence and maintenance of cooperation by natural selection is an enduring conundrum in evolutionary biology, which has been studied using a variety of game theoretical models inspired by different biological situations. The most widely studied games are the Prisoner's Dilemma, the Snowdrift game and by-product mutualism for pairwise interactions, as well as Public Goods games in larger groups of interacting individuals. Here, we present a general framework for cooperation in social dilemmas in which all the traditional scenarios can be recovered as special cases. In social dilemmas, cooperators provide a benefit to the group at some cost, while defectors exploit the group by reaping the benefits without bearing the costs of cooperation. Using the concepts of discounting and synergy for describing how benefits accumulate when more than one cooperator is present in a group of interacting individuals, we recover the four basic scenarios of evolutionary dynamics given by (i) dominating defection, (ii) coexistence of defectors and cooperators, (iii) dominating cooperation and (iv) bi-stability, in which cooperators and defectors cannot invade each other. Generically, for groups of three or more interacting individuals further, more complex, dynamics can occur. Our framework provides the first unifying approach to model cooperation in different kinds of social dilemmas. PMID- 16242729 TI - The emergence of wave emitting centres in an excitable medium. AB - The response of an excitable biological medium to a double local stimulus is considered within the context of a mathematical model for a layer of starving cells of Dictyostelium discoideum, with both spatially one- and two-dimensional (1D and 2D) system being investigated. In contrast to the response usually seen in excitable media, whereby each superthreshold stimulus delivered to the relaxed medium results in the initiation of just one travelling wave, a source emitting a sequence of waves can develop in the present excitable medium after the second stimulus. In a 1D system, only transient wave sources forming a limited number of waves are found. In 2D systems, a permanent wave sources consisting in a pair of spirals are observed as well as the transient wave sources forming circular wave patterns. The general features of the medium dynamics that underlie the observed responses to the double stimulus are discussed. PMID- 16242730 TI - Nonlinear dynamical model and response of avian cranial kinesis. AB - All modern birds have kinetic skulls in which the upper bill can move relative to the braincase, but the biomechanics and motion dynamics of cranial kinesis in birds are poorly understood. In this paper, we model the dynamics of avian cranial kinesis, such as prokinesis and proximal rhynchokinesis in which the upper jaw pivots around the nasal-frontal (N-F) hinge. The purpose of this paper is to present to the biological community an approach that demonstrates the application of sophisticated predictive mathematical modeling tools to avian kinesis. The generality of the method, however, is applicable to the advanced study of the biomechanics of other skeletal systems. The paper begins with a review of the relevant biological literature as well as the essential morphology of avian kinesis, especially the mechanical coupling of the upper and lower jaw by the postorbital ligament. A planar model of the described bird jaw morphology is then developed that maintains the closed kinematic topology of the avian jaw mechanism. We then develop the full nonlinear equations of motion with the assumption that the M. protractor pterygoideus and M. depressor mandibulae act on the quadrate as a pure torque, and the nasal frontal hinge is elastic with damping. The mechanism is shown to be a single degree of freedom device due to the holonomic constraints present in the quadrate-jugal bar-upper jaw-braincase quadrate kinematic chain as well as the quadrate-lower jaw-postorbital ligament braincase-quadrate kinematic chain. The full equations are verified via simulation and animation using the parameters of a Grey Heron (Ardea cinerea). Next we develop a simplified analytical model of the equations by power series expansion. We demonstrate that this model reproduces the dynamics of the full model to a high degree of fidelity. We proceed to use the harmonic balance technique to develop the frequency response characteristics of the jaw mechanism. It is shown that this avian cranial kinesis model exhibits the characteristics of a hardening Duffing oscillator. Beyond the identification of the characteristics of the underlying dynamics, which provides insight into the behavior of the system, the model and methodology presented here provides other potential benefits. A framework has been developed that could be utilized to study the biomechanics of feeding and bite force as well the effects of cranial kinesis on the frequency and modulation of bird songs. PMID- 16242732 TI - Oxidized phospatidylcholine but not native phosphatidylcholine inhibits inducible nitric oxide synthase in RAW 264.7 macrophages. AB - This study was designed to compare the effects of oxidized 1-palmitoyl-2 arachidonyl-sn-glycero-3-phosphorylcholine (PAPC) and native PAPC on the inducible nitric oxide synthase (iNOS) in the macrophage cell line RAW 264.7. Macrophages stimulated by bacterial lipopolysaccharide (1 microg/ml) were incubated with increasing amounts of native or oxidized PAPC (oxPAPC, 10-20 microg/ml). Cells incubated with oxPAPC showed a dose-dependent inhibition of inducible nitric oxide synthesis, as well as reduced iNOS protein expression and mRNA levels. Additionally, chromatin immunoprecipitation assay revealed that oxPAPC reduced the interaction of the active NF-kappaB subunit p65 with the iNOS promoter region when compared to native PAPC. PMID- 16242731 TI - Effects of prenatal cocaine, morphine, or both on postnatal opioid (mu) receptor development. AB - We studied the effects of prenatal cocaine and morphine given separately and in combination on the (1) postnatal brain mu-opioid receptor development and (2) interaction of dopamine with mu receptors. Pregnant rats received single daily intraperitoneal (I.P.) injections of saline, cocaine (20 mg/kg), morphine (2 mg/kg), or the combination of both drugs from day 13 to day 20 of gestation. Postnatal days (P) 1, 7, 14, and 28, whole brains were analyzed for opioid receptor binding and mu mRNA. Prenatal cocaine administered by itself had no significant effect on the ontogeny of brain mu receptors on all the days studied when compared to controls. The morphine-treated group showed a significant increase in mu receptor binding on P1 and P7. Exposure to both cocaine and morphine showed a significant increase in mu receptor density on P1 and P7. In addition, there was also a significant increase in MOR mRNA in both the morphine alone and combination groups. Pretreatment with dopamine D2 receptor antagonist (sulpiride, 20 mg/kg) prior to drug administration showed decreased mu receptor binding on P1 and P7. These results suggest that prenatal exposure to morphine or a combination of cocaine and morphine significantly increases mu receptor density. By P14, mu-opioid receptor binding was no longer different than the control. This may suggest that the effect on receptor may be short-lived and that other key intracellular events may be activated to mediate the long-term effects. Also, the data show that dopaminergic mechanisms are (or opioid-dopamine interaction is) involved in the effects of morphine alone or morphine in combination with cocaine on mu receptor regulation. PMID- 16242733 TI - The influence of season on oxidant-antioxidant status in trained and sedentary subjects. AB - The association between oxidative stress and cardiovascular diseases is a widely accepted fact today. Generally, men have a higher risk of cardiovascular incidents and mortality from acute myocardial infarction and strokes. We have examined sport-associated circannual rhythms of oxidant and antioxidant processes by measuring plasma LPO, erythrocyte SOD, CAT, Gpx activity and plasma hormonal status in both sedentary and long-term trained men and women. We have shown seasonal variations in both oxidant and antioxidant status in all examined groups. The largest difference was observed in the oxidant status between sedentary men and women during autumn and winter, which is considered a period of high coronary risk for men. Sport decreased LPO in trained men in autumn, while the same effect in trained women was shifted towards summer. These data state that regular, long-term physical exercise training induces adaptive responses that confer protection against oxidative stress, as well as the beneficial effect of exercise with regard to season, particularly in men during a period of high coronary risk (autumn and winter, respectively) and in women during summer. PMID- 16242735 TI - Epidemiologic aspects of overweight and obesity in the United States. AB - National survey data from the U.S. show that the prevalence of overweight and obesity among adults remained relatively constant over the 20-year period from 1960 to 1980, began to increase around the mid-1980s and has continued to increase. Data for children and adolescents, based on different definitions, show the same pattern. It can sometimes be more useful to look at the whole distribution of body mass index, rather than on prevalence estimates based on pre defined cutoffs. Data from several countries suggest that for both adults and children, the distribution of BMI has become more skewed over time. Although many hypotheses have been put forward about the causes of the increases, data to address these issues are sparse. Obesity is a well-known risk factor for numerous health conditions. Nonetheless, the health consequences of the increases in obesity have not been fully delineated. Increases in diabetes have been noted in conjunction with the rise in obesity. On the other hand, declines in some other cardiovascular risk factors have been seen at all BMI levels. Rising life expectancy and decreasing heart disease mortality rates seem to confound some of the expectations about the effects of increasing obesity on mortality. The effects of obesity on morbidity may be greater than its effects on mortality. The increasing prevalence of obesity poses challenges for researchers and for policy makers. PMID- 16242736 TI - Anthocyanins from red wine--their stability under simulated gastrointestinal digestion. AB - The stability of anthocyanins from red wine was assessed using an in vitro digestion system that simulated the physiochemical changes that occur in the upper gastrointestinal tract. Anthocyanins in red wine were stable to gastric conditions whereas there was a small loss in total phenol content. After pancreatic digestion, the total anthocyanins were very poorly recovered compared to the bulk phenols in the IN sample, which was previously described as the "serum-available" fraction, and the majority of the anthocyanins and phenols were recovered in the OUT fraction, previously described as the "colon-available" fraction. Removing alcohol from the wine samples prior to the procedure did not markedly affect this pattern. The composition of anthocyanins in the post gastric, IN and OUT samples was analysed using liquid chromatography mass spectrometry. The red wine used contained over 20 identifiable anthocyanins of which the main components were 3-O-glucosides of malvidin, peonidin, petundin, delphidin and cyanidin. Coumaroylated-glucoside derivatives of malvidin, petundin, peonidin, and delphinidin were observed and acetylated glucosides of peonidin, petundin and malvidin were also identified. Anthocyanins with modified aglycones similar to vitisin A derivatives of delphinidin, peonidin, petunidin and malvidin were also identified. After the in vitro digestion procedure, only five anthocyanins could be detected in the IN (serum-available) and the OUT (colon-available) fractions, which were confirmed as malvidin-3-O-glucoside and the vitisin A adducts of malvidin-3-O-glucoside, malvidin-3-O-acetylglucoside, malvidin-3-O-coumaroylglucoside and peonidin-3-O-glucoside. Malvidin-3-O glucoside was recovered at 0.2% in the IN fraction and 0.9% in the OUT fraction. However, the vitisin derivatives were much more stable to pancreatic digestion. Assuming that the vitisin A derivatives display similar biological properties to their parent anthocyanins, their enhanced gastrointestinal stability could lead to enhanced bioavailability and bio-effectiveness in vivo. PMID- 16242737 TI - Negative ion electrospray mass spectrometry of neoflavonoids. AB - The electrospray ionisation mass spectra of the neoflavanoids brazilin and hematoxylin are reported in both their reduced (1 and 2, respectively) and their oxidised forms (3 and 4, respectively). In the reduced forms, breakdown pathways under collision induced decomposition (CID) conditions produce fragments characteristic of rings A and C; in their oxidised forms, the fragments are characteristic of rings B and D. The structural assignments of the fragments are substantiated by recording the spectra after deuterium exchange at the hydroxyl groups. PMID- 16242738 TI - Phomoxins B and C: polyketides from an endophytic fungus of the genus Eupenicillium. AB - Chemical investigations of the culture broth from an endophytic fungus Eupenicillium sp. have afforded two natural products phomoxins B (1) and C (2) as well as the previously reported fungal metabolite eupenoxide (3). Compounds 1 and 2 both contain a cyclic carbonate moiety that is rare among natural products. This paper reports the full spectroscopic characterisation of phomoxins B (1) and C (2) by NMR, UV, IR and MS data. All compounds were inactive against a panel of nosocomial microbes. PMID- 16242739 TI - In vitro metabolism of ginsenosides by the ginseng root pathogen Pythium irregulare. AB - The role of ginseng saponins (ginsenosides) as modulators or inhibitors of disease is vague, but our earlier work supports the existence of an allelopathic relationship between ginsenosides and soilborne microbes. Interestingly, this allelopathy appears to significantly promote the growth of the important ginseng pathogen, Pythium irregulare while inhibiting that of an antagonistic non pathogenic fungus, Trichoderma hamatum. Herein we report on the apparent selective metabolism of 20(S)-protopanaxadiol ginsenosides by an extracellular glycosidase from P. irregulare. Thus, when P. irregulare was cultured in the presence of a purified (> 90%) ginsenoside mixture, nearly all of the 20(S) protopanaxadiol ginsenosides (Rb1, Rb2, Rc, Rd, and to a limited extent G-XVII) were metabolized into the minor ginsenoside F2, at least half of which appears to be internalized by the organism. No metabolism of the 20(S)-protopanaxatriol ginsenosides (Rg1 and Re) was evident. By contrast, none of the ginsenosides added to the culture medium of the non-pathogenic fungus T. hamatum were metabolized. The metabolism of 20(S)-protopanaxadiol ginsenosides by P. irregulare appears to occur through the hydrolysis of terminal monosaccharide units from disaccharides present at C-3 and/or C-20 of ginsenosides Rb1, Rc, Rb2, Rd and G-XVII to yield one major product, ginsenoside F2 and one minor product (possibly G-III). A similar transformation of ginsenosides was observed using a crude protein preparation isolated from the spent medium of P. irregulare cultures. PMID- 16242740 TI - Mefloquine toxicity presenting with polyneuropathy - a report of two cases in India. AB - We report two cases of people from different states in India who developed polyneuropathy following intake of mefloquine for treatment of malaria. To the best of our knowledge this has not been reported before. PMID- 16242741 TI - Kinetics of sedimentation rate, viral load and TNF-alpha in relation to HIV co infection in tuberculosis. AB - The kinetics of potential surrogate markers in HIV-positive (HIV+) and HIV negative (HIV-), smear-positive tuberculosis (Tb+) patients in Gondar, Ethiopia (n = 60) was investigated. Clinical symptoms, sputum conversion, sedimentation rate (SR), HIV viral load and serum levels of TNF-alpha were determined before and 8 weeks after treatment initiation. The co-infection rate of HIV was 45%. There were significantly higher initial levels of SR and TNF-alpha in HIV+/Tb+ patients (79 +/- 29 mm/h and 13.5 +/- 7.6 pg/ml), than in HIV-/Tb+ patients (60 +/- 23 mm/h and 6.8 +/- 5.9 pg/ml, P<0.001). In HIV-/Tb+ patients, there was a marked decrease in SR compared with co-infected patients (46% [33 +/- 24 mm/h at week 8] vs. 24% [61 +/- 27 mm/h at week 8]). The HIV viral load (4.99 [range 3.70 5.92] to 4.90 [range 3.96-5.78] log10 copies/ml from week 0 to 8) and TNF-alpha (13.5 +/- 7.6 to 12.0 +/- 6.0 pg/ml) remained high in HIV+/Tb+ patients. In Tb patients, SR was significantly increased in HIV+ compared with HIV- patients. Additionally, TNF-alpha and HIV viral load remained elevated in HIV+/Tb+ patients following treatment despite clinical improvement comparable to HIV-/Tb+ patients. PMID- 16242743 TI - Transport on fractal river networks: application to migration fronts. AB - A quantitative model of the US colonization in the 19th century is presented. We explore the idea that landscape heterogeneities should have strongly affected this process, as the need for water made the colonizers follow the routes of main rivers and set up their towns near them. So, we study transport processes on fractal networks modeling river basins, a case which may have a great ecological relevance for the study of hydrochory, and in general for species which spread along corridors. The analytical reaction-diffusion model presented here allows to predict the propagation rate of fronts spreading through Peano-like basins, and comparison with the Optimal Channels Network model is also reported. Finally, the propagation rates observed are compared with the results from our model, concluding that migration through fractal media, in spite of being a mathematical idealization of the problem, mimics the dynamics of real systems reasonably well. PMID- 16242742 TI - Syntheses of 19-[O-(carboxymethyl)oxime] haptens of epipregnanolone and pregnanolone. AB - O-(Carboxymethyl)oximes 1 and 2 derived from two epimeric 5beta-pregnanolones (3beta-hydroxy-5beta-pregnan-20-one and 3alpha-hydroxy-5beta-pregnan-20-one) in position 19 were prepared. Two synthetic routes were employed, both using protection of the 20-keto group after reduction into the (20R)-alcohol in the form of acetate. In the first route, (20R)-19-hydroxy-5beta-pregnan-3beta,20-diyl diacetate (3) was transformed into the corresponding 19-[O-(carboxymethyl)oxime] methyl ester 6, then deacetylated by acid and partially silylated with tert butyldimethylsilyl chloride. The desired 3-O-silylated derivative 8 was separated, oxidized to the 20-ketone and protecting groups were sequentially removed to give the first title hapten 1. The second route started from (20R)-19 hydroxy-3-oxopregn-4-en-20-yl acetate (11), which was hydrogenated in the presence of base to the 5beta-pregnan-3-one derivative 12, protected in position 19 with tert-butyldimethylsilyl group and reduced with borohydride. The prevailing 3alpha-alcohol 15 was separated, protected in position 3 with a methoxymethyl group, deprotected in position 19 and transformed into the 19-[O (carboxymethyl)oxime] 19. After deacetylation, esterification with diazomethane and oxidation in position 20, the pregnanolone skeleton was regenerated. Final deprotection steps gave the second title hapten 2. Both haptens, i.e., (19E) 3beta- and -3alpha-hydroxy-20-oxo-5beta-pregnan-19-al 19-[O (carboxymethyl)oxime], were designed for the development of immunoassays of the corresponding parent neuroactive steroids. PMID- 16242744 TI - Parvovirus uncoating in vitro reveals a mechanism of DNA release without capsid disassembly and striking differences in encapsidated DNA stability. AB - The uncoating mechanism of parvoviruses is unknown. Their capsid robustness and increasing experimental data would suggest an uncoating mechanism without capsid disassembly. We have developed an in vitro system to detect and quantify viral DNA externalization and applied the assay on two parvoviruses with important differences in capsid structure, human B19 and minute virus of mice (MVM). Upon briefly treating the capsids to increasing temperatures, the viral genome became accessible in its full-length in a growing proportion of virions. Capsid disassembly started at temperatures above 60 degrees C for B19 and 70 degrees C for MVM. For both viruses, the externalization followed an all-or-nothing mechanism, without transitions exposing only a particular genomic region. However, the heat-induced DNA accessibility was remarkably more pronounced in B19 than in MVM. This difference was also evident under conditions mimicking endosomal acidification (pH 6.5 to 5), which triggered the externalization of B19 DNA but not of MVM-DNA. The externalized ssDNA was a suitable template for the full second-strand synthesis. Immunoprecipitation with antibodies against conformational epitopes and quantitative PCR revealed that the DNA externalized by heat was mostly dissociated from its capsid, however, the low pH-induced DNA externalization of B19 was predominantly capsid-associated. These results provide new insights into parvovirus uncoating suggesting a mechanism by which the full length viral genome is released without capsid disassembly. The remarkable instability of the encapsidated B19 DNA, which is easily released from its capsid, would also explain the faster heat inactivation of B19 when compared to other parvoviruses. PMID- 16242745 TI - Transactivation of the simian varicella virus (SVV) open reading frame (ORF) 21 promoter by SVV ORF 62 is upregulated in neuronal cells but downregulated in non neuronal cells by SVV ORF 63 protein. AB - Simian varicella virus (SVV) infection in primates closely resembles varicella zoster virus (VZV) infection in humans. SVV ORF 63 has 51.6% homology at the amino acid level to VZV ORF 63. We cloned SVV ORFs 63 and 62, transcribed and translated in vitro, and immunoprecipitated the expected proteins with rabbit polyclonal antibodies. Immunoprecipitation analysis revealed that SVV ORF 63 is expressed as a 43-kDa phosphorylated protein in virus-infected cells. In both neuronal and non-neuronal cells, SVV ORF 62 protein alone upregulated the SVV 21 promoter, while SVV ORF 63 protein alone did not have any effect. SVV ORF 62 mediated transactivation of the SVV ORF 21 promoter was upregulated in neuronal cells, but downregulated in non-neuronal cells, by SVV ORF 63 protein. This is the first study in which a varicella protein (ORF 63) expressed during latency has been shown to have a differential effect on a promoter that is also active during latency, in neuronal as compared to non-neuronal cells. PMID- 16242746 TI - Kinetics and functional studies on interaction between the replicase proteins of Tomato Bushy Stunt Virus: requirement of p33:p92 interaction for replicase assembly. AB - The assembly of the functional replicase complex via protein:protein and RNA:protein interactions among the viral-coded proteins, host factors and the viral RNA on cellular membranes is a key step in the replication process of plus stranded RNA viruses. In this work, we have characterized essential interactions between p33:p33 and p33:p92 replication proteins of Tomato bushy stunt virus (TBSV), a tombusvirus with a non-segmented, plus-stranded RNA genome. Surface plasmon resonance (SPR) measurements with purified recombinant p33 and p92 demonstrate that p33 interacts with p92 in vitro and that the interaction requires the S1 subdomain, whereas the S2 subdomain plays lesser function. Kinetic SPR analyses showed that binding of S1 subdomain to the C-terminal half of p33 takes place with moderate binding affinity in the nanomolar range whereas S2 subdomain binds to p33 with micromolar affinity. Using mutated p33 and p92 proteins, we identified critical amino acid residues within the p33:p92 interaction domain that play essential role in replication and the assembly of the tombusviral replicase. In addition, we show that interaction takes place between replication proteins of TBSV and the closely related Cucumber necrosis virus but not between TBSV and the more distantly related Turnip crinkle virus, suggesting that selective protein interactions might prevent the assembly of chimeric replicases carrying replication proteins from different viruses during mixed infections. PMID- 16242747 TI - A rotavirus strain isolated from pig-tailed macaque (Macaca nemestrina) with diarrhea bears a P6[1]:G8 specificity. AB - A distinct rotavirus strain (PTRV) was isolated in cell cultures from a stool sample obtained from a diarrheic 3-year-old female pig-tailed macaque (Macaca nemestrina) that was born at the breeding colony of the University of Washington in Seattle. Unlike other known simian rotavirus strains including vervet monkey rotavirus SA11 which bears P5B[2]:G3 or P6[1]:G3 specificity, rhesus monkey rotavirus MMU18006 with P5B[3]:G3 specificity, pig-tailed macaque rotavirus YK-1 with P[3]:G3 specificity and rhesus monkey rotavirus TUCH with P[24]:G3 specificity, the cell-culture-grown PTRV strain was shown to bear P6[1]:G8 specificity as determined by VP4 (P)- and VP7 (G)-specific neutralization assays as well as gene sequence analyses. The virus in the original diarrhea stool was also shown to bear genotypes P[1] and G8. In addition, the PTRV strain exhibited a "long" electropherotype, subgroup I specificity and NSP4 genotype A specificity. The PTRV probe formed (i) 8-9 hybrid bands with genomic RNAs of various bovine rotavirus strains and (ii) only 2-3 hybrid bands with simian rotavirus RNAs as demonstrated by RNA-RNA hybridization, suggesting a possible bovine origin of the virus. Serologic analysis of serum samples obtained from selected pig-tailed macaques in the colony suggested that a rotavirus bearing P[1]:G8 specificity was endemic among macaques for at least 8 years (1987-1994). This is the first report describing an isolation of a simian rotavirus bearing a non-G3 VP7 and possibly a P6[1] specificities. Because of its unique simian serotype, this virus may prove to be valuable in challenge studies in a non-human primate model in studies of rotavirus immunity. PMID- 16242748 TI - Reorganization of RNA polymerase II on the SV40 genome occurs coordinately with the early to late transcriptional switch. AB - The pattern of organization of RNA polymerase II (RNAPII) in wild-type and mutant cs1085 SV40 chromosomes isolated between 30 min and 48 h post-infection was determined using a combination of chromatin immunoprecipitation (ChIP) techniques. During the course of a wild-type infection, we observed a slow but significant decline in the relative occupancy of RNAPII at the early region and a corresponding increase in occupation in the late region. In the promoter, occupancy began high, decreased to a minimum at 8 h post-infection, and then increased to a high level by 48 h post-infection. In the mutant cs1085, which does not down-regulate early transcription, we observed high occupancy of the early region and the promoter throughout the infection. The changing organization of RNAPII on the wild-type SV40 but not the mutant cs1085 genome appears to be a result of the switch from early to late transcription. PMID- 16242749 TI - Analysis of HIV-1 subtype B third variable region peptide motifs for induction of neutralizing antibodies against HIV-1 primary isolates. AB - The HIV-1 gp120 V3 loop is a potent inducer of neutralizing antibodies for T cell line adapted-HIV-1, but less so for primary isolates. We hypothesized that peptides representative of the diversity of natural HIV-1 V3 loop variants might capture elements of conserved higher order structures and so stimulate broadly reactive neutralizing antibodies. We designed a panel of 29 subtype B V3 sequences postulated to reflect the range of V3 diversity. These peptides were used to immunize guinea pigs. The most effective peptide (62.19) clustered around the subtype B consensus sequence and induced antibodies that reproducibly neutralized 31% of the subtype B HIV-1 primary isolates evaluated, but exhibited limited cross-neutralization of non-subtype B HIV-1 strains. Taken together, these data demonstrated that the limited neutralization profile of antibodies induced by optimal subtype B V3 motifs likely represents the maximum breadth of neutralization of subtype B HIV-1 primary isolates attainable by anti-V3 peptide antibodies. PMID- 16242750 TI - The visual evoked potential in the mouse--origins and response characteristics. AB - The visual evoked potential (VEP) in the mouse is characterized and compared to responses obtained with the electroretinogram (ERG). The results indicate that: 1, the VEP originates in the visual cortex; 2, the rod and cone pathways contribute separately to the VEP; 3, temporal tuning functions for rod and cone ERGs are low pass and band pass, respectively; VEP tuning functions are both band pass; and 4, VEP acuity is 0.62+/-0.156 cycles/degree. The differences in the spatial and temporal tuning functions obtained from the retina and visual cortex provides a tool to investigate signal processing through the visual system. PMID- 16242752 TI - Shape-specific perceptual learning in a figure-ground segregation task. AB - What does perceptual experience contribute to figure-ground segregation? To study this question, we trained observers to search for symmetric dot patterns embedded in random dot backgrounds. Training improved shape segmentation, but learning did not completely transfer either to untrained locations or to untrained shapes. Such partial specificity persisted for a month after training. Interestingly, training on shapes in empty backgrounds did not help segmentation of the trained shapes in noisy backgrounds. Our results suggest that perceptual training increases the involvement of early sensory neurons in the segmentation of trained shapes, and that successful segmentation requires perceptual skills beyond shape recognition alone. PMID- 16242751 TI - Impact of aging and age-related maculopathy on inactivation of the a-wave of the rod-mediated electroretinogram. AB - This study examined the impact of aging and age-related maculopathy (ARM) on the inactivation of phototransduction in rod photoreceptors by measuring the recovery of the a-wave using a paired flash electroretinogram technique. Measurements were made on 32 older adults in normal retinal health, 25 with early ARM, 7 with late ARM, and 20 young adults for comparison purposes. ARM presence and severity were defined by the Wisconsin Age-Related Maculopathy Grading System based on grading of fundus photographs. The inactivation of rod phototransduction exhibited an aging-related slowing. Those with early ARM did not exhibit inactivation slowing over and above what would be expected based on normal retinal aging. Persons in the late stages of ARM exhibited dramatic slowing in inactivation kinetics. PMID- 16242753 TI - Partitioning of dissolved chlorinated ethenes into vegetable oil. AB - Food-grade soybean oil (SoyOil) has been used to enhance in situ anaerobic bioremediation at sites contaminated with chlorinated ethenes (CEs). The abiotic interactions of SoyOil with the CEs may be significant and need to be better understood. The oil: water partition coefficients (Kp) of dissolved CEs into SoyOil were measured in batch tests and ranged from 22 to 1200 with increasing chlorination. CE mixtures significantly reduced the Kp for tetrachloroethene (PCE), but not the other CEs. Simple flow tests were used to quantify the mass transfer coefficients (kL) of dissolved CEs into SoyOil. Higher kL values corresponded to the CEs with higher diffusivity in water. CE mixtures reduced the kL for all of the CEs. The results can be used to predict abiotic interactions and distribution of contaminant mass expected after SoyOil injection, and thus provide a more accurate estimate of the mass of CEs removed due to enhanced biodegradation. PMID- 16242754 TI - Evaluation of redox indicators for determining sulfate-reducing and dechlorinating conditions. AB - An in situ methodology based on covalently bonded redox indicators has been developed for determining when sulfate-reducing conditions exist in environmental samples. Three immobilized redox indicators [thionine (Thi, formal potential at pH 7 (E(0')7) equals 52 mV), cresyl violet (CV, E(0')7 = -81 mV), and phenosafranine (PSaf, E(0')7 = -267 mV)] were tested for their response to sulfide in synthetic solutions and under sulfate-reducing conditions in wastewater slurries. The byproduct of the sulfate-reducing process, sulfide, was found to couple well to CV in the concentration range of 1-100 microM total sulfide ([S(-II)]) and the pH range of 6-8. Thi, the indicator with the highest formal potential, reacts rapidly with sulfide at levels well below 1 microM while PSaf, the indicator with the lowest formal potential, does not couple to sulfide at levels in excess of 100 microM [S(-II)]. The degree of reduction of the indicators (i.e., the fraction of cresyl violet oxidized) in contact with a given level of sulfide can be modeled qualitatively with an equilibrium expression for [S(-II)]-indicator based on the Nernst equation assuming that rhombic sulfur is the product of sulfide oxidation. In a groundwater sample with dechlorinating microbes, reduction of Thi and partial reduction of CV correlated with dechlorination of TCE to cis-DCE. PMID- 16242755 TI - Human foot bones from Klasies River main site, South Africa. AB - The caves at Klasies River contain abundant archaeological evidence relating to human evolution in the late Pleistocene of southern Africa. Along with Middle Stone Age artifacts, animal bones, and other food waste, there are hominin cranial fragments, mandibles with teeth, and a few postcranial remains. Three foot bones can now be added to this inventory. An adult first metatarsal is similar in size and discrete anatomical features to those from Holocene burials in the Cape Province. A complete and well-preserved second metatarsal is especially long and heavy at midshaft in comparison to all Holocene and more recent South African homologues. A large fifth metatarsal is highly distinctive in its morphology. In overall size, these pedal elements resemble specimens from late Pleistocene sites in western Asia, but there are some differences in proportions. The fossils support earlier suggestions concerning a relatively high level of sexual dimorphism in the African Middle Stone Age population. Squatting facets on the two lateral metatarsals appear to indicate a high frequency of kneeling among members of this group. The new postcranial material also underlines the fact that the morphology of particular skeletal elements of some of the 100,000-year-old Klasies River individuals falls outside the range of modern variation. PMID- 16242756 TI - Development of a multiple objective planning theory and system for sustainable air quality monitoring networks. AB - Air quality monitoring data are important bases for air quality management strategies planning and performance assessment. Therefore, the environmental protection authorities need to plan the air quality monitoring network effectively. However, in Taiwan, the national Environmental Protection Administration (EPA) and some county environmental protection bureaus (EPB) separately installed their own monitoring stations. This study developed an integrated methodology and computer system for planning air quality monitoring networks. The environmental, social, and economic objectives and sub-objectives, and their weights were identified using system analysis and multiple objective planning, based on the principles of sustainable development. A multiple objective optimization model and procedure for sustainable air quality monitoring networks planning are developed in this study. According to the procedure, a multiple objective planning system for sustainable air quality monitoring networks (MOPSSAQMN) is developed using computer software based on the modified bounded implicit enumeration algorithm with the constraint arrangement method. The air quality monitoring network of Taoyuan County, in northern Taiwan, was used as a case study to demonstrate the proposed method. Two satisfactory alternatives based on different conditions were generated using MOPSSAQMN. The compared results show that this study generated better alternatives than the current monitoring network. An installation schedule for the alternative was proposed, and its first step is now being implemented by the EPB of Taoyuan County Government. The procedure and computer system developed in this study can be used to assist the competent authorities to devise good and different alternatives for air quality monitoring networks planning. PMID- 16242757 TI - A simulation-based interval two-stage stochastic model for agricultural non-point source pollution control through land retirement. AB - This study presents a simulation-based interval two-stage stochastic programming (SITSP) model for agricultural non-point source (NPS) pollution control through land retirement under uncertain conditions. The modeling framework was established by the development of an interval two-stage stochastic program, with its random parameters being provided by the statistical analysis of the simulation outcomes of a distributed water quality approach. The developed model can deal with the tradeoff between agricultural revenue and "off-site" water quality concern under random effluent discharge for a land retirement scheme through minimizing the expected value of long-term total economic and environmental cost. In addition, the uncertainties presented as interval numbers in the agriculture-water system can be effectively quantified with the interval programming. By subdividing the whole agricultural watershed into different zones, the most pollution-related sensitive cropland can be identified and an optimal land retirement scheme can be obtained through the modeling approach. The developed method was applied to the Swift Current Creek watershed in Canada for soil erosion control through land retirement. The Hydrological Simulation Program FORTRAN (HSPF) was used to simulate the sediment information for this case study. Obtained results indicate that the total economic and environmental cost of the entire agriculture-water system can be limited within an interval value for the optimal land retirement schemes. Meanwhile, a best and worst land retirement scheme was obtained for the study watershed under various uncertainties. PMID- 16242758 TI - The role of web-based environmental information in urban planning--the environmental information system for planners. AB - The Environmental Information System for Planners (EISP) is a proof of concept web-based system designed to support decision making within the UK planning framework by making information on environmental issues more widely accessible. It incorporates relevant outputs from the Natural Environment Research Council (NERC) Urban Regeneration and the Environment (URGENT) research programme and from research directly commissioned by the Office of the Deputy Prime Minister (ODPM). It supports three principal planning functions carried out by local authorities: pre-planning enquiries, development control decisions and strategic planning. Eleven environmental science themes are incorporated: Air quality, Shallow undermining, Landslide susceptibility, Groundwater protection, Flood risk, Drainage, Land contamination, Proximity to landfill, Biodiversity, Natural and Man-made heritage. Decision flow diagrams represent detailed analysis of workflow in each theme, taking account of best practice, regulatory responsibilities and planning guidance. Industry-standard web technologies integrate the flows and provide access to the system via secure web pages. Underpinning the system is an environmental geographical information system (GIS) containing up-to-date data, information and models relevant to each theme. The modular system design allows new legislation and local priorities and datasets to be easily incorporated. Web technology delivers information and research data that have hitherto been difficult for the non-specialist to access and have therefore been under-exploited. The study has demonstrated a successful application of the principles of e-Governance in an area where informed decisions commonly require specialist information. The system, if rolled out nationally, offers potential economic benefits and efficiency savings for both planners and developers. PMID- 16242759 TI - The associations between knowledge, CA125 preoccupation, and distress in women with epithelial ovarian cancer. AB - OBJECTIVE: The goal of the current study was to determine how much women with ovarian cancer know about their cancer and CA125 testing and how much women focus on or are preoccupied with their CA125 levels. We also examined the direct and moderating effects of knowledge and CA125 preoccupation on two measures of distress (depressive symptoms and anxiety). METHODS: Participants were 126 women with epithelial ovarian cancer. Patients completed questionnaires of knowledge, CA125 preoccupation, and distress at the beginning of the first or a new round of chemotherapy. RESULTS: Both knowledge and preoccupation with CA125 levels were associated with depressive and anxiety symptoms. More importantly, knowledge appeared to moderate the association between CA125 preoccupation and depressive symptoms. That is, for patients with lower levels of knowledge, more CA125 preoccupation was associated with more depressive symptoms. However, there was no such association in women with higher levels of knowledge. CONCLUSION: Our results suggest that it may be possible to decrease depressive symptoms in women who are overly preoccupied or worried about their CA125 levels by improving their knowledge of aspects of their disease and the surveillance process. PMID- 16242760 TI - Expression and localization of aquaporin-5 in the epithelial ovarian tumors. AB - OBJECTIVE: To investigate the expression and localization of aquaporin-5 (AQP5) in epithelial ovarian tumors and its clinic significance. METHODS: The expression of AQP5 protein and mRNA in 65 cases epithelial ovarian tumors and 13 cases normal tissue were measured by immunohistochemical technique, Western blotting and RT-PCR, respectively. RESULTS: AQP5 is mainly localized in the basolateral membranes of benign tumor cells, the apical and basolateral membrane of borderline cells and scattered in the membrane of malignant cells and almost no or weak staining in normal ovarian epithelium. The AQP5 expression in ovarian malignant and borderline tumors was significantly higher than that of benign tumors (P < 0.05) and normal tissue (P < 0.05). Of all the epithelial ovarian malignant tumors, the AQP5 expression in cases with ascites volume more than 1000 ml was higher than that of ascites volume less than 500 ml (P < 0.05). Increased AQP5 protein level was associated with lymph node metastasis (P < 0.05). There is a positive correlation between ascites amount and the expression of AQP5 protein and mRNA (P < 0.05), as well as lymph node metastasis and the expression of AQP5 protein and mRNA (P < 0.05). The AQP5 expression was not related with FIGO stage, grade and histological type (P > 0.05). CONCLUSION: The data suggest that overexpression of AQP5 play an important role in tumorigenesis of epithelial ovarian tumors, which may be related to the ascites formation of ovarian carcinoma. PMID- 16242763 TI - Regression and resurgence of the CL following PGF2alpha treatment 3 days after ovulation in mares. AB - The present study was designed to characterize and compare the physiology and ultrasonographic morphology of the corpus luteum (CL) during regression and resurgence following a single dose of native prostaglandin F2alpha (PGF) given 3 days after ovulation, with a more conventional treatment given 10 days after ovulation. On the day of pre-treatment ovulation (Day 0), horse mares were randomly assigned to receive PGF (Lutalyse; 10 mg/mare, i.m.) on Day 3 (17 mares) or Day 10 (17 mares). Beginning on either Days 3 or 10, follicle and CL data and blood samples were collected daily until post-treatment ovulation. Functional and structural regression of the CL in response to PGF treatment were similar in both the Day 3 and 10 groups, as indicated by an abrupt decrease in circulating concentrations of progesterone, decrease in luteal gland diameter and increase in luteal tissue echogenicity. As a result, the mean +/- S.E.M. interovulatory interval was shorter (P < 0.0001) in the Day 3 group (13.2 +/- 0.9 days) than in the Day 10 group (19.2 +/- 0.7 days). Within the Day 3 group, functional resurgence of the CL was detected in 75% of the mares (12 of 16) beginning 3 days after PGF treatment, as indicated by transient major (6 mares) and minor (6 mares) increases (P < 0.05 and < 0.1, respectively) in progesterone. Correspondingly, mean length of the interovulatory interval was longer (P < 0.03) in mares with major resurgence (15.8 +/- 1.6 days) than in mares with minor (11.2 +/- 1.2 days) and no resurgences (13.5 +/- 0.3 days) in progesterone. Structural resurgence of the CL in the Day 3 group and functional and structural resurgence in the Day 10 group were not detected. In conclusion, PGF treatment 3 days after ovulation resulted in structural and functional regression of the CL and hastened the interval to the next ovulation, despite post-treatment resurgences in progesterone. PMID- 16242761 TI - Guillain-Barre syndrome occurring after adjuvant chemo-radiotherapy for endometrial cancer. AB - BACKGROUND: The association between Guillain-Barre syndrome (GBS) and malignancy is uncommon and has not been previously reported in gynecological cancers. CASE: Our case documents this syndrome occurring in a patient shortly after completion of adjuvant chemo-radiotherapy for endometrial carcinoma. We review the current literature and discuss potential pathogenic mechanisms of this likely paraneoplastic association. CONCLUSION: GBS in cancer patients is a potentially life-threatening condition and should be differentiated from simple chemotherapy toxicity, particularly as effective treatment is available. PMID- 16242762 TI - Detection of damage in mammalian sperm cells. AB - Ejaculated semen is washed for in vitro fertilization or diluted and processed to allow optimal and long-term low temperature liquid- and cryo-preservation. However, sperm are vulnerable to the washing, dilution, temperature and osmotic changes involved in sperm storage. In this review, a number of techniques are considered for detecting damaged spermatozoa. Staining protocols have been developed to detect the membrane and organelle integrity of mammalian sperm cells. Plasma membrane integrity is usually assessed after staining cells with membrane-impermeable dyes or alternatively with acetylated membrane (AM) permeable probes that are selectively de-esterified and become membrane impermeable and thus entrapped into viable cells only (AM ester loading). Organelle-specific dyes are commonly used to detect functionality of mitochondria or the acrosome. A distortion in the lateral and bilayer organization of lipids as well as the peroxidation of fatty acid moieties can be quantified and localized in living sperm. The relation of a disordering in the sperm membrane's lipid architecture and sperm deterioration versus capacitation is discussed. Finally, the integrity of sperm DNA can be measured at three different levels by assessing the degree of DNA-protamine condensation, the incidence of breaks and nicks in the DNA and the frequency of fragmentation of the nuclei into sub haploid apoptotic bodies. The relevance of detecting DNA aberrations and especially the putative link to the incidence of apoptosis is critically considered. PMID- 16242764 TI - Flow cytometric sexing of mammalian sperm. AB - This review reexamines parameters needed for optimization of flow cytometric sexing mammalian sperm and updates the current status of sperm sexing for various species where this technology is currently being applied. Differences in DNA content have provided both a method to differentiate between these sex determining gametes and a method to sort them that can be used for predetermining sex in mammals. Although the DNA content of all cells for each mammalian species is highly conserved, slight but measurable DNA content differences of sperm occur within species even among cattle breeds due to different sizes of Y-chromosomes. Most mammals produce flattened, oval-headed sperm that can be oriented within a sorter using hydrodynamic forces. Multiplying the percentage the difference in DNA content of the X- or Y-chromosome bearing sperm times the area of the flat profile of the sperm head gives a simple sorting index that suggests that bull and boar sperm are well suited for separation in a flow sorter. Successful sperm sexing of various species must take into account the relative susceptibilities of gametes to the stresses that occur during sexing. Sorting conditions must be optimized for each species to achieve acceptable sperm sexing efficiency, usually at 90% accuracy. In the commercial application of sperm sexing to cattle, fertility of sex-sorted bull sperm at 2 x 10(6)/dose remains at 70-80% of unsexed sperm at normal doses of 10 to 20 x 10(6) sperm. DNA content measurements have been used to identify the sex-chromosome bearing sperm populations with good accuracy in semen from at least 23 mammalian species, and normal-appearing offspring have been produced from sexed sperm of at least seven species. PMID- 16242765 TI - Which variability? AB - Drai and Grodzinsky provide a valuable analysis that offers a way of disentangling the effects of Movement and Mood in agrammatic comprehension. However, their mathematical implementation (Beta model) hides theoretically relevant information, i.e., qualitative heterogeneities of performance within the patient sample. This heterogeneity is crucial in the variability debate. PMID- 16242766 TI - The belief in a just world and subjective perceptions of society: a developmental perspective. AB - The hypothesis guiding this study stated that just world beliefs (i.e., the belief that the world is orderly and just) are primitive beliefs that lose their importance across age as they become replaced by more sophisticated forms of reasoning enabling individuals to handle a world that is neither orderly nor just. In addition, just world beliefs were thought to relate to perceptions of inequality and collectivism within society. In this study, a cross-sectional design was employed involving 235 secondary school pupils and 268 psychology students divided over six age groups with mean ages 12, 14, 16, 18, 20, and 22 years and older. They were presented with the Just World Beliefs Scale and the Individualism-Collectivism Scale. Outcomes revealed that general beliefs in a just world begin to loose their importance around the age of 12, followed by personal beliefs around the age of 16. Vertical collectivism related positively to general and personal just world beliefs showing that the experience of social 'inequality' plays an important role in the maintenance of such beliefs. PMID- 16242767 TI - Binding of wogonin to human gammaglobulin. AB - The binding of wogonin to HGG was studied by spectroscopic method including circular dichroism (CD), fourier transformation infrared spectra (FT-IR), fluorescence spectra. The binding parameters and the thermodynamic parameters for the reaction have been calculated according to Sips method and Gibbs-Helmholtz equation, respectively, at different temperatures. AutoDock3.05 program was used to calculate the interaction modes between the drug and HGG. The Sips plots indicated that the binding of HGG to wogonin at 297, 304, 310 and 317 K is characterized by two binding sites with the average affinity constant Ko at 2.102x10(4), 2.078x10(4), 1.956x10(4) and 1.931x10(4), respectively. The binding process was exothermic, spontaneous and entropy driven, as indicated by the thermodynamic analyses, and the major part of binding energy is electrostatic interaction accompanied by hydrophobic interaction and hydrogen bond. The secondary structure compositions of free HGG and its wogonin complexes were estimated by the FT-IR spectra and the curve-fitted results of amide I band, which are in good agreement with the analyses of CD spectra. Furthermore, the average binding distance between wogonin and HGG (5.60 nm) was obtained on the basis of the theory of Forster energy transfer. PMID- 16242768 TI - Bioluminescent imaging: emerging technology for non-invasive imaging of bone tissue engineering. AB - Bone tissue engineering is a multidisciplinary research area in which new strategies are developed to treat patients with large bone defects as occurring during e.g. hip revisions, upon trauma or in spinal fusions. In vivo evaluation of bone formation in animal models is highly relevant for graft evaluation but is time-consuming, invasive and difficult to quantify. As a consequence, most in vivo studies ignore the dynamic nature of bone regeneration and the molecular processes underlying it. In vivo bioluminescent imaging (BLI) is a relatively young research field with great potential that may overcome these problems. BLI encompasses non-invasive imaging of luciferase gene activity using cooled charge coupled device cameras in luciferase transgenic animals or in grafted, luciferase transgenic cells. The imaging procedure is technically simple and quantifiable. Because luciferase expression can be put under the control of tissue-specific regulatory elements, BLI allows non-invasive imaging of processes highly relevant to bone tissue engineering like differentiation, apoptosis, vasculogenesis and inflammation. In this review, we describe the basic principle of BLI and discuss transgenic animals and constructs currently available for application in bone tissue engineering. Furthermore, we reflect on technical developments that will make BLI even more promising for future application in bone tissue engineering research. PMID- 16242769 TI - Micropatterned cell co-cultures using layer-by-layer deposition of extracellular matrix components. AB - Micropatterned cellular co-cultures were fabricated using three major extracellular matrix components: hyaluronic acid (HA), fibronectin (FN) and collagen. To fabricate co-cultures with these components, HA was micropatterned on a glass substrate by capillary force lithography, and the regions of exposed glass were coated with FN to generate cell adhesive islands. Once the first cell type was immobilized on the adhesive islands, the subsequent electrostatic adsorption of collagen to HA patterns switched the non-adherent HA surfaces to adherent, thereby facilitating the adhesion of a second cell type. This technique utilized native extracellular matrix components and therefore affords high biological affinity and no cytotoxicity. This biocompatible co-culture system could potentially provide a new tool to study cell behavior such as cell-cell communication and cell-matrix interactions, as well as tissue-engineering applications. PMID- 16242770 TI - The placenta cytokine network and inflammatory signals. AB - Throughout its entire lifespan, the placenta is able to produce as well as respond to a variety of inflammatory stimuli. Many signaling molecules and concurrent pathways responsible for the propagation of an inflammatory response have been identified in placental cells. From early developmental stages onward, the secretory activity of placenta cells clearly contributes to increase local as well as systemic levels of cytokines and inflammatory molecules. Two aspects of the progression of an immune response have been particularly investigated: the highly regulated process of invasion and implantation and, the induction of preterm labor associated with infections. With the progression of pregnancy, the physiological role of most placental cytokines is more uncertain. Many placental cytokines are similar to adipose tissue derived cytokines. One possibility is that they contribute to the low grade systemic inflammation developing during the third trimester of pregnancy. The prevalent hypothesis is that activation of some inflammatory pathways is necessary to induce maternal insulin resistance which is required for the progression of normal gestation. As an integrative organ, the placenta may relay or enhance the contribution made by the cells of the adipose tissue and immune system in non-pregnant individuals. In pregnancy complicated with obesity or diabetes mellitus, continuous adverse stimulus is associated with dysregulation of metabolic, vascular and inflammatory pathways supported by increased circulating concentration of inflammatory molecules. It is believed that maternal adipose tissue and placental cells both contribute to the inflammatory situation by releasing common molecules. For example, the accumulation of leptin and TNF-alpha is associated with an increased production for markers of inflammation, fibrotic response, vascular remodeling and proteins facilitating lipid storage within the placenta. PMID- 16242771 TI - Confocal laser scanning microscope study of terminal villi vessels in normal term and pre-eclamptic placentas. AB - The aim of this study was to compare immunocytochemical confocal scanning laser microscopy measures of villus capillarization in control placentas with pre eclamptic ones. Accordingly, placentas from normal term pregnancies (n=3) and cases of late-onset pre-eclampsia without intrauterine growth retardation (IUGR) featuring normal uterine artery Doppler (n=3) were analyzed by confocal scanning laser microscopy (CSLM), which is a powerful technique for obtaining three dimensional reconstructions of any kind of blood vessels (arteries, veins, capillaries). A laser light beam is used in order to detect CD34 antibody-related immunofluorescence, which is a marker of endothelial cells. Villus capillarization was assessed by estimating the following parameters: number of pixels, mean, maximum and minimum immunofluorescence amplitude. Our results show a significant hyper-ramification of the capillary loop in pre-eclamptic placentas, featuring irregular profile and narrow lumina. Such findings support the hypothesis that several agents causing angiogenesis and vasoconstriction affect villus vessels in pre-eclamptic placentas, thus promoting a lasting condition of fetal hypoxia by decreasing endothelial surface and materno-fetal exchanges. PMID- 16242773 TI - Mitochondria play a critical role in shaping the exocytotic response of rat pancreatic acinar cells. AB - We have previously demonstrated [M. Campos-Toimil, T. Bagrij, J.M. Edwardson, P. Thomas, Two modes of secretion in pancreatic acinar cells: involvement of phosphatidylinositol 3-kinase and regulation by capacitative Ca(2+) entry, Curr. Biol. 12 (2002) 211-215] that in rat pancreatic acinar cells, Gd(3+)-sensitive Ca(2+) entry is instrumental in governing which second messenger pathways control secretory activity. However, in those studies, we were unable to demonstrate a significant increase in cytoplasmic [Ca(2+)] during agonist application as a result of this entry pathway. In the present study, we combined pharmacology with ratiometric imaging of fura-2 fluorescence to resolve this issue. We found that 2 microM Gd(3+) significantly inhibits store-mediated Ca(2+) entry. Furthermore, both the protonophore, CCCP (5 microM) and the mitochondrial Ca(2+)-uptake blocker, RU360 (10 microM), led to an enhancement of the plateau phase of the biphasic Ca(2+) response induced by acetylcholine (1 microM). This enhancement was completely abolished by Gd(3+); and as has been previously shown for Gd(3+), RU360 led to a switch to a wortmannin-sensitive form of exocytosis. Using MitoTracker Red staining we found a close association of mitochondria with the lateral plasma membrane. We propose that in rat pancreatic acinar cells, capacitative Ca(2+) entry is targeted directly to mitochondria; and that as a result of Ca(2+) uptake, these mitochondria release "third" messengers which both enhance exocytosis and suppress phosphatidylinositol 3-kinase-dependent secretion. PMID- 16242774 TI - Homodimerization and internalization of galanin type 1 receptor in living CHO cells. AB - Galanin is a 29- to 30-aa-long neuropeptide affecting feeding, cognitive, and sexual behavior. It exerts its effects through galanin receptors 1, 2 and 3, which are all seven transmembrane domain G-protein coupled receptors (GPCRs). The GPCRs have been shown to function as monomers, homodimers, heterodimers and oligomers. In this study, we examined the extent of galanin receptor 1 (GalR1) dimerization and internalization in living CHO cells using fluorescence resonance energy transfer (FRET) and time lapse confocal imaging. Ratio imaging analysis and emission spectral analysis revealed substantial homodimerization of GalR1. In addition, internalization of GalR1 after 1h of agonist stimulation with the GalR1 agonist galanin (1-29) was observed with time lapse fluorescence imaging, whereas stimulation with the GalR2 specific agonist galanin (2-11) did not lead to internalization. Treatment of GalR1 transfected cells with the non-selective adenylyl cyclase activator forskolin influenced the rate of internalization when administered together with galanin (1-29). These results indicate that GalR1 can act as a dimer on the cell surface and that receptor desensitization and internalization was observed after stimulation with the agonist galanin (1-29). Western blots further confirm the FRET data that GalR1-XFP dimerizes and can be detected in the cell as a monomer or dimer using antibodies to XFP. Internalization and dimerization of GalR1 is shown, contributing to the regulation of galanergic signaling. PMID- 16242775 TI - WT1 gene expression in children with Down syndrome and transient myeloproliferative disorder. AB - Transient myeloproliferative disorder (TMD) is found in 10% of newborns with Down syndrome (DS). Myeloid leukemia develops in 25% within the following 3 years. Little is known about markers predicting leukemia occurrence. We studied expression levels of the Wilms tumor gene (WT1) by real-time quantitative PCR (RQ PCR) in peripheral blood of five infants with TMD. WT1 levels were elevated similar to findings in AML. Longitudinal studies showed normalization of the WT1 level in all patients except one who developed GATA1 mutated myeloid leukemia at 11 months of age. The lack of normalization of WT1 level may be a predictor of leukemia development and WT1 expression may be an attractive marker for monitoring of minimal residual disease. PMID- 16242776 TI - Transforming growth factor-beta1 inhibits all-trans retinoic acid-induced apoptosis. AB - The interaction between retinoids and transforming growth factor-beta1 (TGF beta1) leading to regulation of proliferation, differentiation and apoptosis is not still fully understood. In this study, we demonstrated that a combination treatment with all-trans retinoic acid (ATRA) and TGF-beta1 led to the enhancement of ATRA-induced suppression of cell proliferation, which is accompanied by inhibition of ATRA-induced apoptosis in human leukemia HL-60 cells. This effect was preceded by the arrest of cells in G0/G1 cell cycle phase linked with pRb protein dephosphorylation, continuous accumulation of p21 and transiently increased level of p27, inhibitors of cyclin-dependent kinases. Inhibition of ATRA-induced apoptosis by TGF-beta1 was associated with an increased level of Mcl-1 protein, an anti-apoptotic member of Bcl-2 family, but not with inhibition of mitochondrial membrane depolarization. Levels of other Bcl 2 family proteins (Bcl-2, Bcl-X(L), Bad, Bak, Bax) were unaffected by simultaneous ATRA and TGF-beta1 treatment, when compared to ATRA alone. Upregulation of c-FLIP(L) protein, an inhibitor of apoptosis induced by tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), correspond with inhibition of ATRA-induced (autocrine TRAIL-mediated) caspase-8 activation and apoptosis. These results suggest that apoptosis inhibition associated with proliferation block could depend on modulation of the TRAIL apoptotic pathway and regulation of the Mcl-1 protein level. In summary, we demonstrate that the balance of processes leading to regulation of proliferation and differentiation of myeloid cells can modulate cell sensitivity to apoptosis-inducing stimuli. PMID- 16242777 TI - Viscum album agglutinin-I (VAA-I) induces apoptosis and degradation of cytoskeletal proteins in human leukemia PLB-985 and X-CGD cells via caspases: lamin B1 is a novel target of VAA-I. AB - Viscum album agglutinin-I (VAA-I) is a potent inducer of cell apoptosis and possesses anti-tumoral activity. Using PLB-985 and chronic granulomatous disease (X-CGD) cells, which lack expression of gp91(phox), VAA-I was found to induce apoptosis in both cell lines as assessed by cytology, DNA laddering and degradation of the cytoskeletal protein gelsolin. Both cell lines expressed caspase-3 and -8 and VAA-I activated these caspases. We demonstrated that lamin B(1) is a novel target to VAA-I and its degradation was reversed by a pan-caspase inhibitor and by a caspase-6, but not a caspase-8, inhibitor. PMID- 16242778 TI - Using synthetic miRNA mimics for diverting cell fate: a possibility of miRNA based therapeutics? PMID- 16242779 TI - Herpetic eye disease in diabetic patients. AB - PURPOSE: To study the incidence of herpetic eye disease (HED) of the ocular surface in diabetics. DESIGN: Observational historical cohort study. SETTING: A district of the largest health maintenance organization in Israel (the Central District of Clalit Health Services). PARTICIPANTS: We reviewed the electronic medical records of all patients older than 50 years (159634 patients) in the district, and of these, 22382 (14.0%) patients had diabetes mellitus. METHODS: All filled prescriptions for acyclovir eye ointment between January 1, 2001 and December 31, 2003 (1483 tubes) and all hemoglobin A1c laboratory tests during 2003 (41910 tests) were documented. An ocular surface HED event was defined when a patient consumed at least 1 tube of topical acyclovir per month, whereas no acyclovir use was documented 3 months before and 3 months after that event. MAIN OUTCOME MEASURES: Incidence of ocular surface HED events in diabetics compared with nondiabetics adjusted for age and gender. RESULTS: After age and gender adjustment, significantly more diabetics had ocular surface HED (5.21 per thousand) compared with nondiabetics (4.27 per thousand; P<0.0001). Stratification by age revealed a significantly higher prevalence of HED in diabetics, aged 60 to 79 years. Recurrent herpetic events occurred during the study period in 25.2% of HED-affected diabetics, and in 16.6% of HED-affected nondiabetics (P = 0.05). Diabetics with poor glycemic control (mean annual hemoglobin A1c > 9%) consumed significantly more ocular acyclovir (P = 0.01). Multivariate analysis revealed this effect to be independent of age, gender, place of birth, or place of residency. CONCLUSIONS: Ocular surface HED is significantly more common among patients with diabetes mellitus. Poor glycemic control correlates with increased consumption of ocular acyclovir in diabetic patients. PMID- 16242780 TI - Comparison of fluorescence polarization assay with card and complement fixation tests for the diagnosis of goat brucellosis in a high-prevalence area. AB - An evaluation of fluorescence polarization assay (FPA) to detect antibodies against Brucella melitensis according to the Mexican Official Norm (NOM) was performed. In this study, a total of 2582 goat serum samples from a high prevalence area in northeast Mexico where vaccination is applied, were used. Of these, 1094 were classified as NOM negatives (card test (CT) negatives or CT positives/complement fixation test (CFT) negatives) and 1488 as NOM positives (CT and CFT positives). The receiver operator characteristics (ROC) curve analysis was used to obtain the FPA sensitivity (83.5%), specificity (82.2%) and accuracy (88.2%) compared with NOM criteria, using a cut-off value of 89mP for positive samples. In addition, FPA produced 84.1% of negative results versus 65.7% of CT using 1094 CFT negative samples, which indicated that FPA performance was better than CT to detect negative samples or differentiate samples from vaccinated animals. Finally, FPA showed 95.8% sensitivity when using 702 negative non vaccinated samples. Taken together, these results suggested that FPA might replace CT as a screening test for its better performance compared with CFT, its adjustable cut-off useful in different epidemiological situations, and for its reliability, ease of performance, comparable cost with CT regimen, and potential application in field and high-throughput laboratories. The use of FPA as screening test will help to reduce the percentage of goats wrongly slaughtered because of brucellosis misdiagnosis. More studies on FPA are required for its approval as diagnostic tool for goat brucellosis. PMID- 16242781 TI - Prophylactic, therapeutic and anti-metastatic effects of an HPV 16mE6Delta/mE7/TBhsp70Delta fusion protein vaccine in an animal model. AB - Human papillomaviruses (HPVs), particularly HPV-16, are not only causally linked to cervical cancers but also play an important role in the development of other cancers. The oncoproteins, E6 and E7, are consistently coexpressed in the majority of HPV-containing carcinomas and their metastatic lesions, and are critical to the induction and maintenance of malignant phenotype, and also can cause tumor metastasis. Therefore, E6 and E7 represent ideal tumor-specific antigens for the development of immunotherapy to prevent and treat HPV-associated cancers and their metastases. The powerful antigenic nature of Mycobacterium tuberculosis heat shock protein 70 (TBhsp70) is emphasized by evidence that mammals are capable of recognizing murine and human multiple B and T cell epitopes in this protein, and therefore allows it to be used as an adjuvant-free carrier to stimulate the immune response to a covalently linked fusion partner. In our present study, we developed a recombinant TBhsp70Delta protein expression vector that permits the production of other protein fused to TBhsp70Delta. A recombinant HPV-16mE6Delta/mE7/TBhsp70Delta fusion protein was expressed and purified, and immunization with the fusion protein in the absence of adjuvant was capable of providing strong protection to C57BL/6 mice against challenge and rechallenge with TC-1 cells, but not HPV negative Lewis lung cancer cells, and induced established TC-1 tumor regression and led to long-term survival. Consistent with the in vivo results, the fusion protein immunization in the absence of adjuvant induced cytolytic T lymphocytes recognized specifically TC-1 tumor cells in vitro. We also demonstrated that immunization with the fusion protein in the absence of adjuvant was effective in both preventing and treating TC-1 metastatic lesions in the lung metastasis model. In particular, immunization with the fusion protein caused regression of established lung metastatic lesions in 50% of immunized animals. This study represents an instance of tumor therapy with a TBhsp70Delta fusion protein and provides the scientific basis for the clinical application of the HPV16mE6Delta/mE7/TBhsp70Delta fusion protein in the treatment of HPV-associated cancers and their metastases. PMID- 16242782 TI - Toll-like receptor 2-dependent and -independent activation of macrophages by group B streptococci. AB - Group B streptococcus (GBS), a capsulated gram-positive bacterium, is a major cause of newborn infections. Although the innate immune receptor Toll-like receptor (TLR) 2 has been shown to primarily recognize gram-positive bacterial products, the production of TNF by macrophages treated with heat-killed GBS (HK GBS) does not depend on TLR2. In this report, we have characterized HK-GBS induced activation of macrophages derived from wildtype and TLR2-deficient mice. Microarray analysis demonstrated that HK-GBS activation of macrophages induces both TLR2-independent and -dependent signals. While the expression of a major fraction of genes in macrophages induced by HK-GBS does not depend on TLR2, induction of several important molecules involved in host innate immunity such as IL-6, IL-1beta, and lipocalin 2 is severely impaired in the absence of TLR2 signaling. Furthermore, we show that HK-GBS utilizes centrifugation sensitive components to induce rapid activation of TLR2(-/-) macrophages and that HK-GBS induced activation of macrophages is not mediated through its genomic DNA. Together, our results demonstrate that HK-GBS induces TLR2-dependent antimicrobial gene activation and provide further understanding of the molecular basis of host innate response to GBS infection. PMID- 16242783 TI - Darkness induced neuroplastic changes in the serotoninergic system of the chick retina. AB - Sensory experience is critical for the formation of neuronal circuits and it is well known that neuronal activity plays a crucial role in the formation and maintenance of synapses. In the vertebrate retina, exposure to different environmental conditions results in structural, physiological, neurochemical and pharmacological changes. Serotoninergic (5HT) amacrine cells of the chicken retina are bistratified interneurons whose primary dendrites descend through the inner nuclear layer (INL) to branch in the inner plexiform layer (IPL) forming two plexi, an outer network, localized to sublamina 1, and an inner network, localized to sublamina 4 and 5 of the IPL. Their development is temporally correlated with the establishment of synapses in the retina and with the emergence of the typical adult electroretinogram. It is unknown, however, which role these cells play in processing visual information and whether visual deprivation modifies their phenotype. Here, we show that, in the chicken, red light rearing from hatching to postnatal day 12 significantly alters the stratification pattern of 5HT amacrine cells, inhibiting their age-dependent pruning measured with morphometric and densitometric procedures; as well as increasing serotonin immunoreactivity measured as relative optical density. This change in dendritic arborization, accompanied by an increase in serotonin concentration in dark adapted conditions, may decrease visual threshold, thus increasing visual sensitivity. PMID- 16242784 TI - Noradrenaline transporter and its turnover rate are decreased in blood lymphocytes of patients with major depression. AB - Lymphocytes possess transporters of serotonin and dopamine, and also contain monoamines. The objective of this work was to determine the presence of noradrenaline transporters, the turnover rate of noradrenaline and serotonin in lymphocytes of major depression patients, and to correlate the biochemical parameters with the severity of the disorder. Lymphocytes from peripheral blood were isolated by Ficoll/Hypaque, and noradrenaline transporter was studied by binding of [3H]nisoxetine: control group (29, age 31.52+/-1.08, 7 men) and major depression patients (35, age 36.68+/-1.69, 6 men), Hospital Vargas de Caracas. Diagnostic was done by criteria of the American Psychiatric Association and severity by Hamilton Scale for Depression. Levels of noradrenaline, serotonin, 3 methoxy-4-hydroxyphenylglycol and 5-hydroxyindoleacetic acid were determined by HPLC. Turnover rate was evaluated by the ratios of monoamines and metabolites. Correlations were done between the biochemical parameters and the severity of depression. The score of Hamilton for Depression was 22.77+/-0.51. There was a reduction in the number of transporters in lymphocytes of patients, 0.95+/-0.27 versus 4.06+/-1.67 fmol/10(6) cells. Levels of monoamines and metabolites did not significantly differ between patients and controls. However, there was a higher monoamine/metabolite ratio in lymphocytes of patients, indicating a reduction of metabolic turnover rate. Also there was a relative greater concentration of noradrenaline than serotonin in the lymphocytes of the patients, as indicated by the ratio noradrenaline/serotonin. Noradrenergic and serotonergic turnover is decreased in blood peripheral lymphocytes of major depression patients; the reduction in noradrenaline transporter could be related to changes in intracellular levels, and these modifications could result in functional changes of the immune system. PMID- 16242785 TI - A study of the influence of mouth-breathing in some parameters of unstimulated and stimulated whole saliva of adolescents. AB - OBJECTIVE: The aim of this study was to analyze some standard parameters of the unstimulated and stimulated whole saliva of mouth-breathers and a control group to determine if these variables present any difference in mouth-breathers compared to control group, since these parameters of saliva can influence the oral health. METHODS: Saliva samples were collected from 61 adolescents aged 10 19 years; 30 were mouth-breathers and 31 were nose-breathers. The unstimulated salivary specimen was collected, followed by collection of the stimulated saliva. Soon after collecting the salivary sample, the flow rate and buffering capacity were determined. The samples were then stored at -80 degrees C until analysis was performed. The analysis consisted of the determination of protein content and total, free and bound sialic acid levels. RESULTS: No statistically significant difference was observed in the flow rate, buffering capacity, protein content, total and bound sialic acid levels of unstimulated and stimulated saliva, nor in the free sialic acid of stimulated saliva. However, the levels of free sialic acid of the unstimulated saliva were significantly higher in the mouth-breather compared to the control group. CONCLUSIONS: Since a higher level of free sialic acid is indicative of an increase in the number of bacteria in saliva, our findings suggest that mouth-breathers retain more bacteria in oral tissues. PMID- 16242786 TI - Diagnosis of pediatric laryngopharyngeal reflux. PMID- 16242787 TI - Rigid bronchoscopy for the suspicion of foreign body in the airway. AB - OBJECTIVE: Airway foreign bodies present a diagnostic dilemma and has been recognized for many years. Since aspiration of foreign bodies can be a serious and sometimes fatal problem, early intervention and proper management is vital. METHOD: In this retrospective study, the results of 1887 bronchoscopies, which were performed between the years 1973 and 2004 for the suspicion of foreign body aspiration in children, were presented. Various instruments and techniques were used over 31-year period and rigid bronchoscopy was the preferred method of foreign body extraction. RESULTS: There were 1106 boys and 781 girls with the median age of 2.3 years. Seventy-four percent of patients were less than 3 years old. The most common type of foreign body (89.9%) was organic; watermelon seeds (39.7%) were the most frequent organic foreign bodies. Eight hundred and twenty three patients (43.6%) were referred to our clinic within the first 24h of the event while 4.5% of the patients were admitted to the hospital with the suspicion of foreign body in the airway later than one month. At bronchoscopy, a foreign body was identified in 79.1% of patients and no foreign body was seen in 20.9% of patients. Foreign bodies were encountered in 96.3% of the patients with positive history whereas 28.1% of the patients with negative history had foreign body. Of the patients with foreign bodies, 93.2% had positive history. Overall, the positive history was obtained from 85.2% of patients. The incidence of postbronchoscopic tracheotomy, thoracotomy, and overall mortality rate were 0.47, 0.15, and 0.21%, respectively. CONCLUSION: Otolaryngologists should consider foreign body aspiration in the airway in the differential diagnosis of any patient with the complaints of stridor, dyspnea, sudden onset of cough and intractable and recurrent lower respiratory tract infections. A careful history and physical examination were strong indicators of the diagnosis and raised the index of suspicion of an aspirated foreign body. Timely intervention with the experienced surgical team would decrease the complication rate and mortality rate. However, prevention of aspiration with the education of parents and caregivers is very important. PMID- 16242788 TI - Potentiating role of interleukin 2 (IL-2) receptors in the midbrain periaqueductal gray (PAG) upon defensive rage behavior in the cat: role of neurokinin NK(1) receptors. AB - Feline defensive rage is a form of aggression occurring in nature in response to a threatening condition and is elicited under laboratory conditions by electrical stimulation of the medial hypothalamus or midbrain periaqueductal gray (PAG). Since it has recently been shown that cytokines can modulate neurotransmitter release, the present study was designed to determine the effects of administration of interleukin 2 (IL-2) into the PAG upon defensive rage elicited from the medial hypothalamus. Microinjections of relatively low doses of IL-2 into the dorsal PAG significantly facilitated defensive rage behavior elicited from the medial hypothalamus. The specificity of this phenomenon was supported by the following findings: (1) IL-2 induced effects were dose- and time-dependent, (2) the facilitative effects of IL-2 could be completely blocked by pre-treatment of the injection site with either anti-IL-2 or anti-IL-2 receptor antibody and (3) IL-2 administration into the PAG showed no effect upon another form of aggression, namely predatory attack, elicited from the lateral hypothalamus. The findings further demonstrated that the effects of IL-2 were mediated by an NK(1) receptor mechanism since pre-treatment of the PAG with an NK(1) receptor antagonist completely blocked the facilitating effects of IL-2. Immunocytochemical observations supported these findings by demonstrating an extensive pattern of labeling of IL-2Ralpha in the dorsal PAG. The present study thus demonstrates that IL-2 in the dorsal PAG potentiates defensive rage behavior and is mediated through an NK(1) receptor mechanism. PMID- 16242789 TI - The electroencephalographic substratum of the awakening. AB - The aim of the present study was to characterize the regional electroencephalographic substratum of the awakening process by means of a Hz-by Hz EEG spectral power analysis. For this purpose, we recorded a group of 25 female subjects who slept for at least two consecutive nights in the laboratory. The post-sleep waking EEG was compared to the one recorded during the presleep wakefulness from four midline derivations (Fz-A1, Cz-A1, Pz-A1, Oz-A1). Results indicated that the first 10 min after awakening are characterized by an increase of EEG power in the low-frequency range (1-9 Hz) compared to the corresponding presleep waking period, and by a significant decrease of EEG power in the beta range (18-24 Hz). As regards topographic differences, the increase of EEG power upon awakening in the delta-theta range showed a parieto-occipital prevalence. Moreover, the occipital derivation showed a larger decrease of power in the beta range as compared to the other derivations. In conclusion, the EEG substratum of the sleep offset period is characterized by a pattern of increased EEG power in the delta-theta and low-alpha bands, and of decreased power in the beta range. This pattern could be considered as the spectral EEG signature of the sleep inertia phenomenon. The state of post-sleep EEG hypo-arousal does not subside in the first 10-min period after awakening considered in the present analysis. Finally, according to our results, the more posterior scalp locations show stronger EEG signs of sleep inertia, and could be the last ones to properly wake up. PMID- 16242790 TI - Alterations in cognitive function in prepubertal mice with protein malnutrition: relationship to changes in choline acetyltransferase. AB - We have found that protein malnutrition (PM) causes a significant impairment of memory-related behavior on the 15th and 20th day after the start of PM (5% casein) feeding in prepubertal mice but not in postpubertal mice, as measured by a passive-avoidance task. This impairment was almost completely reversed by merely switching to a standard protein (20% casein) diet on the 10th day after the start of PM. However, the reversal was not observed when the switching to a standard protein regimen was done on the 15th day of the PM diet. Interestingly, the impairment of memory-related behavior on the 20th day was improved by the chronic administration of physostigmine (0.1 mg/kg/day x last 10 days, i.p.), a cholinesterase inhibitor. To correlate brain cholinergic neuron function with the memory-related behavior impairment induced by PM, microphotometry was used to determine the histological distribution of the imunofluorescence intensity for choline acetyltransferase (ChAT), a functional marker of presynapse in cholinergic neurons. The change in the intensity of fluorescence indicated that ChAT protein was decreased in the hippocampus (CA1, CA3 and dentate gyrus) on the 20th day after PM feeding in comparison with controls. These results suggest the possibility that the memory-related behavior deficits observed in prepubertal mice with PM are caused by a dysfunction of the cholinergic neurons in the hippocampus. PMID- 16242791 TI - Adenylyl cyclase activity and its modulation in the gills of Mytilus galloprovincialis exposed to Cr6+ and Cu2+. AB - The adenylyl cyclase (AC)/cAMP system regulates a large number of physiological functions in bivalve mussels, although its basal properties and the potential effects of environmental pollutants are scarcely studied. We characterized some properties of AC and measured both the enzyme activity and the cAMP levels in the gills of the filter-feeding sea mussel Mytilus galloprovincialis. Basal AC activity was 5.6+/-0.8 pmol cAMP 10 min(-1) mg protein(-1) and showed a Km value of 0.82+/-0.06 mM for ATP in the presence of 5mM Mg2+. It was stimulated up to 2.5- and 3.5-fold by 5-HT and GTPgammaS, respectively. Similarly to what was found in other bivalves, forskolin is a poor activator that reached significant stimulation only at 100 microM. Both basal and 5-HT-stimulated AC activity were significantly increased in the gills of mussels exposed for 7 days in aquaria to Cr6+ (10 ng/l) and Cu2+ (5 microg/l). The cAMP content of the gill under these conditions was also significantly higher than in control animals. In vitro exposure of gill membrane preparations to Cr6+ and Cu2+ induced a bimodal effect. Cu2+ significantly stimulated AC activity at nanomolar concentrations, but a strong inhibition was displayed in the micromolar range. A similar bell-shaped curve was obtained in the presence of Cr6+, with maximal AC stimulation at 10( 8)M and inhibition at 10(-5)M. Overall, these data suggest that the mussel AC/cAMP system can be affected with different patterns by heavy metals. AC activity is strongly affected by acute exposure to heavy metals in vitro, probably through a direct interaction of the pollutants with the enzyme molecule, while AC activity and cAMP content increase in organisms exposed for 7 days in vivo, probably as a defense response to acclimate the physiological functions to the environmental challenge. PMID- 16242793 TI - Characterisation of a secreted N-acetyl-beta-hexosaminidase from Trichinella spiralis. AB - A thorough investigation was conducted for glycoside hydrolase activities in the secreted proteins of Trichinella spiralis. The data demonstrated that the only secreted glycosidase with significant activity was an exo-beta-hexosaminidase with catalysis of the substrates N-acetyl-beta-D-glucosamine, N-acetyl-beta-D galactosamine and N-acetyl-beta-D-glucosamine-6-sulphate proceeding with an efficiency similar to the human isozyme beta-hexosaminidase A (Hex A). The hydrolysis of N-acetyl-beta-D-glucosamine followed Michaelis-Menten kinetics with a K(m) of 0.187+/-0.025 mM, and catalysis was inhibited competitively by both N acetyl-beta-d-glucosamine and N-acetyl-beta-D-galactosamine, with K(i) values of 15.75+/-0.99 and 1.17+/-0.24 mM, respectively. The enzyme was maximally active at pH 4.4, had a temperature optimum at 54 degrees C and was thermolabile. We observed no cleavage of N-acetylglucosamine beta1-4 linkages in N acetylchitooligosaccharides, but significant hydrolysis of N-acetylglucosamine beta1-2 linked to mannose in glycans was detected indicating that the secreted enzyme is linkage specific. The enzyme was partially purified and identified by SDS-PAGE and Western blotting as a protein with an apparent molecular mass of 50 kDa. We established that the protein was glycosylated and showed that the glycan was decorated with tyvelose (3,6-dideoxy-D-arabino-hexose). Matrix-assisted laser desorption/ionisation mass spectrometry (MALDI-MS) analysis demonstrated that the carbohydrate moeity was a tyvelose capped tetra-antennary N-glycan corresponding to the structure Tyv(4)Fuc(5)HexNAc(10)Hex(3). All our studies suggest that this is a novel variant of a secreted N-acetyl-beta-hexosaminidase. PMID- 16242792 TI - Antioxidant systems and lipid peroxidation in Bathymodiolus azoricus from Mid Atlantic Ridge hydrothermal vent fields. AB - Enzymatic defenses involved in protection from oxygen radical damage were determined in gills and mantle of Bathymodiolus azoricus collected from three contrasting Mid-Atlantic Ridge (MAR) hydrothermal vent fields (Menez-Gwen, Lucky Strike and Rainbow). The activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidases (GPx) (total and Se-dependent), and levels of total oxyradical scavenging capacity (TOSC), metallothioneins (MT) and lipid peroxidation (LPO) were determined in B. azoricus tissues and the impact of metal concentrations on these antioxidant systems and lipid peroxidation assessed. SOD, CAT, TOSC, MTs and LPO levels were higher in B. azoricus gills while glutathione peroxidases (total and Se-dependent) were higher in the mantle, and with the exception of CAT, were of the same order of magnitude as in other molluscs. TOSC levels from Menez-Gwen indicate that the vent environment at this site is less stressful and the formation of ROS in mussels is effectively counteracted by the antioxidant defense system. TOSC depletion indicates an elevated ROS production in molluscs at the other two vent sites. Cytosolic SOD, GPx and LPO were more relevant at Lucky Strike (Bairro Alto) where levels of essential (Cu and Zn) and toxic metals (Cd and Ag) were highest in the organisms. CAT activity and LPO were predominant at the Rainbow vent site, where an excess of Fe in mussel tissues and in vent fluids (the highest of all three vent sites) may have contributed to increased LPO. Therefore, three distinct pathways for antioxidant enzyme systems and LPO based on environmental metal speciation of MAR vent fields are proposed for Bathymodiolus gills. At Menez-Gwen, TOSC towards peroxyl and hydroxyl radicals and peroxynitrite are predominant, while at Lucky Strike cytosolic SOD activity and GPx are the main antioxidant mechanisms. Finally at Rainbow, catalase and lipid peroxidation are dominant, suggesting that resistance of mussels to metal toxicity at these vent fields decreases in the sequence Menez Gwen > Lucky Strike and Rainbow. PMID- 16242794 TI - Angiotensin type 1 receptor is linked to inhibition of nitric oxide production in pulmonary endothelial cells. AB - We previously demonstrated that angiotensin II (Ang II) stimulates an increase in nitric oxide synthase (NOS) mRNA levels, eNOS protein expression and NO production via the type 2 (AT2) receptor, whereas signaling via the type 1 (AT1) receptor negatively regulates NO production in bovine pulmonary artery endothelial cells (BPAECs). In the present study, we investigated the components of the AT1 receptor-linked signaling pathway(s) that are involved in the downregulation of eNOS protein expression in BPAECs. Treatment of BPAECs with either AT1 receptor antagonists or an anti-AT1 receptor antibody induced eNOS protein expression. Furthermore, intracellular delivery of GP-Antagonist-2A, an inhibitor of Galphaq proteins, and treatment of BPAECs with U73122, a phosphatidylinositol-phospholipase C (PLC)-specific inhibitor, enhanced eNOS protein expression. Treatment of BPAECs with the cell-permeable calcium chelator, BAPTA/AM, increased eNOS protein expression at 8 h, while increasing intracellular calcium with either thapsigargin or A23187 prevented Ang II-induced eNOS protein expression. Phorbol myristate acetate (PMA), a protein kinase C (PKC) activator, completely prevented Ang II-stimulated eNOS protein expression at 8 h, whereas depletion of PKC by long-term treatment with PMA, induced eNOS protein expression. Treatment of BPAECs with a PKCalpha-specific inhibitor or transfection of BPAECs with an anti-PKCalpha neutralizing antibody stimulated eNOS protein expression. Conversely, rottlerin, a PKCdelta specific isoform inhibitor had no effect on basal or Ang II-stimulated eNOS protein expression. Moreover, treatment of BPAECs with U73122, BAPTA/AM and PKCalpha-specific inhibitors increased NO production at 8 h. In conclusion, Ang II downregulates eNOS protein expression via an AT1 receptor-linked pathway involving Galphaq/PLC/calcium/PKCalpha signaling pathway in BPAECs. PMID- 16242795 TI - Regulation of NGF and BDNF by dexamethasone and theophylline in human peripheral eosinophils in allergics and non-allergics. AB - BACKGROUND: Recent studies have shown that the neurotrophins NGF and BDNF are produced by eosinophils. The influence of neurotrophins in allergic diseases including asthma has been described. The regulation by pharmacological substance remains unclear. OBJECTIVES: The aim of this study was to assess whether approved pharmacological substances in the treatment of asthma such as corticosteroids or theophylline regulate neurotrophins on a cellular level. METHODS: Eosinophils were purified by negative immunoselection from allergics and non-allergic donors. Eosinophils were incubated with dexamethasone and theophylline and supernatants were collected for measurement of neurotrophic factors. The content of neurotrophins in eosinophil lysates was determined by ELISA. Regulation of stored NGF and BDNF was demonstrated by Western-blotting and flow cytometry while influence on transcription level was demonstrated by RT-PCR. RESULTS: Eosinophils produce and release the neurotrophins NGF and BDNF at different levels in allergics and non-allergics. Dexamethason lead to a significant downregulation of NGF in eosinophils of allergics. The levels of BDNF were not significantly reduced. Theophylline did not influence the levels of NGF nor BDNF significantly. CONCLUSIONS: The production of the neurotrophin NGF was downregulated by an established substance such as dexamethasone. This might further contribute to the pharmacological potential of corticosteroids in allergic asthma. PMID- 16242796 TI - Early reduction in left ventricular contractile reserve detected by dobutamine stress echo predicts high-dose chemotherapy-induced cardiac toxicity. AB - BACKGROUND: High-dose chemotherapy (HDC) is utilized in high-risk cancer patients. This type of treatment may induce cardiac toxicity which becomes clinically evident weeks or months after HDC. Hence, the possibility of early identification of patients who will develop cardiac impairment is strategic for its clinical implications. The aim of this study was to identify possible early changes of left ventricular contractile reserve (LVCR) in cancer patients undergoing HDC, as well as to evaluate the relevance of such changes as predictors of chemotherapy-induced cardiotoxicity. METHODS: In forty-nine female patients scheduled for HDC, due to poor-prognosis breast cancer, dobutamine stress echocardiography (DSE) was performed, before each of the three HDC cycles (C1, C2, C3), and 1, 4, and 7 months after the end of chemotherapy. According to rest left ventricular ejection fraction (LVEF) evaluated within 18 months after HDC (f-LVEF), patients were allocated to Group A (LVEF < 50% and >10 absolute units reduction) and to Group B (LVEF > or = 50%). RESULTS: Rest LVEF didn't show any significant difference between the two groups except at f-LVEF. Peak LVEF and LVCR significantly decreased in Group A only, starting from C3. At C3, a > or = 5 units fall in LVCR was found to be predictive for f-LVEF drop below 50%. CONCLUSIONS: In patients undergoing HDC, low-dose DSE allows the early identification of patients at a high risk of developing cardiac dysfunction. PMID- 16242797 TI - Losartan reduces monocyte chemoattractant protein-1 expression in aortic tissues of 2K1C hypertensive rats. AB - BACKGROUND: Previous study has demonstrated an arterial inflammatory response in aortic tissues in several hypertensive models which can not be fully explained by hemodynamic forces. This study sought to investigate the effect of angiotensin II (Ang II)and its subtype-1 receptor blocker Losartan on the chemokine expression of monocyte chemoattractant protein-1(MCP-1) in aortic tissues of acute stage of 2-kidney-1-clip (2K1C) hypertensive rats. METHODS: 2K1C renovascular hypertension was produced in male Wistar-Kyoto(WKY) rats by placing a silver clip with an internal diameter of 0.2 mm around the left renal artery. The MCP-1 mRNA on aortic wall was detected by in situ hybridization and reverse transcription polymerase chain reaction(RT-PCR); the levels of Ang II in plasma and aorta were determined by radioimmunoassay. The concentration of MCP-1 in supernatant of cultured endothelial cells (ECV-304) was measured by ELISA. RESULTS: No MCP-1 was exhibited in aortic wall of normotensive rats both by RT-PCR and in situ hybridization; it would be expressed on the aortic wall of rats in 7 days after 2K1C hypertensive model was formed, especially in intima. The expression of MCP-1 in aortic wall was increased with the duration of hypertension and correlated with local Ang II activity (r=0.594, P=0.02) other than those in plasma, it was decreased obviously after being treated by Losartan for 28 days (optical density of MCP-1/GAPDH ratio: 0, 0.58+/-0.10, 1.14+/-0.09, 1.52+/-0.20, 0.66+/-0.07, respectively, P<0.01). Ang II had increased the expression of MCP-1 in endothelial cells; the highest levels had been performed at 1x10(-7)mol/l Ang II or after 4 h, respectively; and losartan markedly reduced the expression of MCP 1. CONCLUSIONS: The expression of MCP-1 significantly increases in aortic tissues of the acute stage 2K1C hypertensive rats and is decreased markedly by treatment of losartan. These findings imply Ang II may be involved in facilitating MCP-1 production in hypertension, and may provide a molecular link between hypertension and the development of atherosclerosis. PMID- 16242798 TI - Analysis of the extension of Q-waves after infarction with body surface map: relationship with infarct size. AB - AIMS: We aimed to characterize the extension of Q-waves after a first ST-segment elevation myocardial infarction using body surface map (BSM) and its relationship with infarct size quantified with cardiovascular magnetic resonance imaging (CMR). METHODS AND RESULTS: Thirty-five patients were studied 6 months after a first ST-segment elevation myocardial infarction (23 anterior, 12 inferior). All cases had single-vessel disease and an open artery. The extension of Q-waves was analyzed by means of a 64-lead BSM. Infarct size was quantified with CMR. Absence of Q-waves in BSM was observed in 5 patients (14%), 2 of whom (40%) had >1 segment with transmural necrosis. Absence of Q-waves in 12-lead ECG was observed in 8 patients (23%), 7 of whom (87%) had >1 segment with transmural necrosis. Patients with inferior infarctions (n=12, 34%) showed a larger number of Q-waves in BSM (18+/-7.1 leads) than patients with anterior infarctions (n=23, 66%; 3.7+/ 3.6 leads; p<0.0001). When the study group was analysed as a whole, the total number of Q-waves detected in BSM did not correlate with the number of necrotic segments (r=0.15; p=0.4). In anterior infarctions, a number of Q-waves >median (2 leads) was related to a higher number of necrotic segments (5.1+/-2.4 vs. 2+/-2.2 segments; p=0.004). The same was observed in inferior infarctions (median 20 leads: 3.5+/-1.9 vs. 1.2+/-1.2 segments; p=0.03). CONCLUSION: In a stable phase after a first ST-segment elevation myocardial infarction, absence of Q-waves does not mean non-transmural necrosis. Using BSM, extension of Q-waves is much higher in inferior infarctions; a separate analysis depending on infarct location is necessary. A major BSM-derived extension of Q-waves is related to larger infarct size both in anterior and in inferior infarctions. PMID- 16242799 TI - A model of contagious bovine pleuropneumonia transmission dynamics in East Africa. AB - The dynamics of contagious bovine pleuropneumonia (CBPP) transmission vary widely between livestock production systems. This paper describes the development of a homogeneous, stochastic, compartmental model for CBPP transmission in pastoral herds of East Africa. The model was built using parameter estimates based on data published in the literature and on observations of livestock owners obtained through participatory research. The basic reproduction number for CBPP in southern Sudan was estimated to range from 3.2 to 4.6. The homogeneous model indicates that the critical community size for the persistence of CBPP falls within the typical herd sizes for pastoral communities in East Africa suggesting that individual isolated herds are capable of maintaining infection indefinitely. Vaccination alone with currently available vaccines was unlikely to eradicate the disease. PMID- 16242800 TI - A heterogeneous population model for contagious bovine pleuropneumonia transmission and control in pastoral communities of East Africa. AB - Pastoral cattle live in highly structured communities characterized by complex contact patterns. The present paper describes a spatially heterogeneous model for the transmission of contagious bovine pleuropneumonia (CBPP) developed specifically for pastoral communities of East Africa. The model is validated against serological data on the prevalence of CBPP infection in several communities of southern Sudan and against livestock owner information on community structure, livestock contact and cattle exchange. The model is used to assess the impact of alternative control strategies including mass and elective vaccination programmes, potential treatment regimes and the combination of vaccination and treatment in a single unified strategy. The results indicate that the eradication of CBPP using mass vaccination with currently available vaccines is unlikely to succeed. On the other hand, elective control programmes based on herd level vaccination, treatment of clinical cases or a combination of both vaccination and treatment enabled individual livestock owners to capture a large benefit in terms of reduced animal-level prevalence and mortality experience. The most promising intervention scenario was a programme which combined the vaccination of healthy animals with treatment of clinical cases. PMID- 16242801 TI - Multiplex PCR for simultaneous detection of five virulence hemolysin genes in Vibrio anguillarum. AB - A multiplex PCR was developed for detection of hemolysin-producing Vibrio anguillarum using primers targeting five hemolysin genes (vah1, vah2, vah3, vah4 and vah5). This method was successful in amplifying reactions containing as little as 100 fg of genomic template DNA. The direct detection of V. anguillarum in clinical specimens by this multiplex PCR was also successful in reactions containing as few as 10 bacterial cells. This multiplex PCR method can be a rapid and sensitive method for detecting pathogenic V. anguillarum. PMID- 16242802 TI - Microarray analysis of Bacillus cereus group virulence factors. AB - Bacillus cereus, B. thuringiensis and B. anthracis are closely related medically and economically important bacterial species that belong to the B. cereus group. Members of the B. cereus group carry genes encoding several important virulence factors, including enterotoxins, phospholipases and exotoxins. Since it is difficult to differentiate among B. cereus group members, and because Bacillus virulence factors are very important for pathogenesis, we explored the use of microarray-based detection of virulence factor genes as a tool for strain identification and for determining virulence. Our method requires an initial multiplex PCR amplification step, followed by identification of the PCR amplicons by hybridization to an oligonucleotide microarray containing genes for all three types of Bacillus virulence factors including B. anthracis virulence factors. The DNA chip described here contains 21 identical arrays used for analysis of seven samples in triplicates. Using the arrays, we found that virulence factors are present in several combinations in the strains analyzed. This work also demonstrates the potential of oligonucleotide microarrays for medical, food safety and biodefense analysis of microbial pathogens. PMID- 16242803 TI - Engineering chromosomes for delivery of therapeutic genes. AB - The ability to create fully functional human chromosome vectors represents a potentially exciting gene-delivery system for the correction of human genetic disorders with several advantages over viral delivery systems. However, for the full potential of chromosome-based gene-delivery vectors to be realized, several key obstacles must be overcome. Methods must be developed to insert therapeutic genes reliably and efficiently and to enable the stable transfer of the resulting chromosomal vectors to different therapeutic cell types. Research to achieve these outcomes continues to encounter major challenges; however recent developments have reiterated the potential of chromosome-based vectors for therapeutic gene delivery. Here we review the different strategies under development and discuss the advantages and problems associated with each. PMID- 16242804 TI - Complete sequence and organization of the human adenovirus serotype 46 genome. AB - Out of 51 human adenoviral serotypes recognized to date, 32 of them belong to species D. Members of species D adenoviruses are commonly isolated from immune suppressed patients (organ transplant) and patients suffering from AIDS. The role of species D adenoviruses in pathogenesis is currently unclear. To derive new insights into the genetic content and evolution of species D adenoviruses and as a first step towards development of human adenovirus serotype 46 (Ad46) as vector, the complete nucleotide sequence of the virus was determined. The size of the genome is 35,178 bp in length with a G+C content of 56.9%. All the early and late region genes are present in the expected locations of the genome. The deduced amino acid sequences of all late region genes, with the exception of fiber, exhibited high degree of homology with the corresponding proteins of other adenoviruses. The deduced amino acid sequences of early regions E1, E3 and E4 showed a high degree of homology with the corresponding proteins of adenoviruses belonging to species D and less homology with the corresponding proteins of adenoviruses of other species. The homologues of Ad5 E3 region genes encoding 12.5K, gp19K, 10.4K, 14.5K and 14.7K are conserved in the genome of Ad46. However, the E3 region of Ad46 lacks genes encoding 6.7K and adenovirus death protein (ADP) but contains two additional open reading frames with a coding capacity of 433 and 281 amino acids. The fiber protein of Ad46 is 200 amino acids smaller than the fiber protein of Ad5 and contains only 10 pseudo-repeats in the shaft region. To facilitate the manipulation of the genome, the complete genome of Ad46 was cloned into a single bacterial plasmid. Following transfection into E1 complementing cell lines, the virus was recovered demonstrating the feasibility of viral genome manipulation for generation of recombinant viruses. PMID- 16242805 TI - Viral RNA in the bloodstream suggests viremia occurs in clinically ill rabies infected mice. AB - Data regarding the occurrence of a viremia during rabies virus infections are contradictory. Here, we attempted to clarify the dissimilar results using a qualitative TaqMan PCR assay to detect viral RNA in blood of mice that had been injected intramuscularly with rabies virus. Viral RNA was detected at two different intervals. Initially, RNA was present in blood of 30/32 (94%) mice, from 1h to 2 days after injection of virus. The RNA in the blood at this time most likely resulted from trauma to blood vessels at the injection site and leakage of the inoculated virus into the circulation. Thereafter, from 3 to 30 days, viral RNA was undetectable in the blood of 37 mice that remained free of clinical disease. However, and more importantly, viral RNA was detected again in 21/25 (84%) mice that became clinically ill and were exsanguinated 2-4 days after the onset of paralysis. The presence of viral RNA in blood of the clinically ill mice might have been due to an escape of virus into the bloodstream as a result of viral replication induced injury in the central nervous system and other tissues. Anti-rabies virus neutralizing antibody was detected in sera of 11/21 (52%) clinically ill mice whose blood was positive for rabies viral RNA. The presence of viral RNA in the bloodstream of mice that developed clinical rabies suggested that a viremia might occur in rabies-infected mice. Thus, the current opinion that a viremia does not occur in experimental or natural rabies infections of other species might need to be re-evaluated. PMID- 16242806 TI - Coping strategies in men and women with type 2 diabetes in Swedish primary care. AB - Based on findings regarding gender differences in the experience and complications of diabetes, we studied coping strategies in men and women with type 2 diabetes in relation to their demographic, medical, socio-economic and psychosocial situation. Altogether 232 Swedish-born type 2 diabetes patients aged 35-64 years at four primary health care centres in Stockholm County were studied, 121 men and 111 women. Coping strategies were assessed by the General Coping Questionnaire (GCG), which describes five orientations dichotomised into positive and negative opposites: self-trust/fatalism; problem focusing/resignation; cognitive revaluation/protest; social trust/isolation; and minimisation/intrusion. Socio-economic and medical data were taken both from a questionnaire and from medical records. Gender differences for the coping strategies resignation, protest and isolation were found (p<0.05), with higher scores for women. The most important medical factor associated with coping strategies was HbA1c. Other significant factors detected in the multivariate analyses were psychiatric disorder, cohabitation and daily smoking. Thus, coping strategies and gender are important factors that should be addressed more in diabetes health care. PMID- 16242807 TI - Demand and supply of emergency help: an economic analysis of Red Cross services. AB - This paper analyzes supply and demand side characteristics of (voluntary) Red Cross services in Austria. The demand side analysis is based on a contingent valuation study on people's willingness to pay for emergency treatment, transportation services and disaster relief activities. The supply side is identified by a high percentage of volunteers in the Red Cross organization which makes the provision of emergency help at low cost possible. We find that aggregate benefits of Red Cross services exceed their cost of production. Policy conclusions are drawn with respect to future recruitment and funding: whereas intrinsic motivation is important for the decision to volunteer, and financial incentives play a minor role in general, the young Red Cross activists work voluntarily for self-realization reasons and to continue their education. Age specific recruitment strategies accompanied by word-of-mouth advertising are recommended to address potential volunteers. As long as the volunteering character of Red Cross services will be maintained and cost of production will not go up an increase of funds does not seem necessary in the future. Moreover, a radical change in the structure of funding may crowd out both donations and voluntary labor supply. PMID- 16242808 TI - Laboratory-scale application of fiber optic transflection dip probe (FOTDP) for in situ monitoring of gas phase ozone in unsaturated porous media. AB - A fiber optic transflection dip probe (FOTDP) system was developed for in situ and real-time monitoring of the transport of gas phase ozone in unsaturated porous media. A unique property of this system is the employment of a dip probe, which is inserted within the porous media. At the probe's tip, incoming light interacts with gas phase ozone and is partially reflected back into the probe by a mirror attached to the tip. Calibration of the FOTDP system was successfully carried out with various ozone concentrations using a column packed with glass beads. The ozone breakthrough curves (BTCs) were obtained by converting normalized UV intensities into gas phase ozone concentrations. The FOTDP system worked well for in situ monitoring of gas phase ozone using a column packed with sand under various water saturations in the presence of SOM and reflected the ideal transport phenomena of gas phase ozone for various flow rates. PMID- 16242809 TI - The Stice model of overeating: tests in clinical and non-clinical samples. AB - The present study tested the dual pathway model of Stice [. A review of the evidence for a sociocultural model of bulimia nervosa and an exploration of the mechanisms of action. Clinical Psychology Review, 14, 633-661 and . A prospective test of the dual-pathway model of bulimic pathology: mediating effects of dieting and negative affect. Journal of Abnormal Psychology, 110, 124-135.] in a non clinical sample of female adolescents and a clinical sample of female eating disorder patients. The model assumes that negative affect and restrained eating mediates the link between body dissatisfaction and overeating. We also tested an extended version of the model postulating that negative affect and overeating are not directly related, but indirectly through lack of interoceptive awareness and emotional eating. Structural equation modelling was used to test our models. First, in the two samples, body dissatisfaction and drive for thinness were associated with overeating/binge eating. In both clinical and adolescent sample, we found support for the negative affect pathway and not for the restraint pathway. Lack of interoceptive awareness and emotional eating appear to (partly) explain the association between negative affect and overeating. Emotional eating was much more strongly associated with overeating in the clinical than in the adolescent sample. In sum, we found substantial evidence for the negative affect pathway in the dual pathway model. The link between body dissatisfaction and overeating in this respect might be explained by the fact that negative affect, due to body dissatisfaction, is related to a lack of awareness of personal feelings and to eating when dealing with negative emotions, which on its turn is associated with overeating. PMID- 16242810 TI - What should be done with nasal Staphylococcus aureus carriers? PMID- 16242811 TI - Reduced pulsatility does not explain renal hyporesponsiveness to cardiac natriuretic peptides in pulmonary hypertension. AB - A pulsatile secretory pattern is assumed to improve hormonal efficiency. We examined the short-term time courses of circulating atrial (ANP) and brain natriuretic peptides (BNP) in patients with pulmonary hypertension (PH) and reduced renal efficiency of ANP-BNP, reflected by low natriuresis/ANP or BNP ratios. Compared to controls, we observed a persistence of ANP and BNP pulsatility in PH patients with a similar periodicity of 20min. Pulse amplitude increased proportionally to the rise in mean plasma level observed in patients (around 27%). In PH patients, the decrease in ANP-BNP renal efficiency is not attributable to a loss of the rhythmic pulsatility of these hormones. PMID- 16242816 TI - Antiretroviral therapy for HIV-1 vaccine efficacy trial participants who seroconvert. AB - Instead of preventing infection, most HIV-1 vaccines in clinical trials are directed at inducing cytotoxic HIV-1-specific T cell (CTL) responses which may control viral replication and subsequently modify the clinical course of HIV-1 infection. Thus, vaccine efficacy trial designs must follow participants who become HIV-infected and monitor the course of HIV-1 infection, in order to assess the effect of vaccination on HIV-1 disease progression. This post-infection evaluation will assess time to reach specific CD4 and viral load thresholds as well as time to initiation of antiretroviral therapy. This paper discusses current literature and guidelines on the initiation of highly active antiretroviral therapy (HAART) for persons who become HIV-infected during HIV-1 vaccine trials, focusing both on acute and early HIV-1 infection, since participants in HIV-1 vaccine and other prevention trials will typically be identified within 3-6 months after HIV-1 acquisition. A standardized HAART protocol for HIV-1 vaccine efficacy trial participants who become HIV-infected is essential to the evaluation of CTL-based HIV-1 vaccines on the natural history of HIV-1 infection. PMID- 16242812 TI - Gene expression correlates of neurofibrillary tangles in Alzheimer's disease. AB - Neurofibrillary tangles (NFT) constitute one of the cardinal histopathological features of Alzheimer's disease (AD). To explore in vivo molecular processes involved in the development of NFTs, we compared gene expression profiles of NFT bearing entorhinal cortex neurons from 19 AD patients, adjacent non-NFT-bearing entorhinal cortex neurons from the same patients, and non-NFT-bearing entorhinal cortex neurons from 14 non-demented, histopathologically normal controls (ND). Of the differentially expressed genes, 225 showed progressively increased expression (AD NFT neurons > AD non-NFT neurons > ND non-NFT neurons) or progressively decreased expression (AD NFT neurons < AD non-NFT neurons < ND non-NFT neurons), raising the possibility that they may be related to the early stages of NFT formation. Immunohistochemical studies confirmed that many of the implicated proteins are dysregulated and preferentially localized to NFTs, including apolipoprotein J, interleukin-1 receptor-associated kinase 1, tissue inhibitor of metalloproteinase 3, and casein kinase 2, beta. Functional validation studies are underway to determine which candidate genes may be causally related to NFT neuropathology, thus providing therapeutic targets for the treatment of AD. PMID- 16242817 TI - Eustachian tube possesses immunological characteristics as a mucosal effector site and responds to P6 outer membrane protein of nontypeable Haemophilus influenzae. AB - The eustachian tube (ET) plays an important role in the pathogenesis of otitis media (OM). To better understand its biology and to develop a nasal vaccine for preventing OM, mucosal lymphocytes in the ET were analyzed, and the ET's immunological function was investigated. Mononuclear cells were isolated from murine ET, and lymphocyte subsets were analyzed by flow cytometry. Antibody producing cells were determined by enzyme-linked immunospot assay. The expression of cytokine mRNA in ET CD4(+) T cells was determined by RT-PCR. Results in naive mice showed that the ET contained many immunocompetent cells, including a relative large number of IgA-producing cells and Th2 cytokine-expressing T cells. Next, we investigated antigen-specific immune responses in the ET. Mice were immunized intranasally with the P6 outer membrane of nontypeable Haemophilus influenzae (NTHi) and cholera toxin (CT), and P6-specific immune responses in the ET were examined. P6-specific IgA producing cells markedly increased in the ET. Moreover, in vitro stimulation with P6 of purified CD4(+) T cells from immunized mice resulted in the proliferation of CD4(+) T cells that expressed Th2 cytokine mRNA. These results indicate that the ET might be characterized as a mucosal effector site and that antigen-specific IgA and Th2 immune responses could be induced in the ET by intranasal immunization. These findings suggest that the ET might be a key immunological organ in the pathogenesis of OM, and in the development of a nasal vaccine. PMID- 16242818 TI - Interferon-gamma production in vitro from whole blood of foot-and-mouth disease virus (FMDV) vaccinated and infected cattle after incubation with inactivated FMDV. AB - Studies were performed to determine whether a rapid method to detect cell mediated immune responses to foot-and-mouth disease virus (FMDV) could be used either as a diagnostic test or provide a correlate of protection in animals post vaccination. Using protocols based on the BOVIGAM assay for tuberculosis, whole blood samples from FMDV vaccinated or control animals, before and after challenge infection, were stimulated overnight with inactivated FMDV antigen. The quantity of interferon gamma (IFN-gamma) produced in the supernatants was measured using an ELISA. Specific induction of IFN-gamma was detected in samples from vaccinated, infected and vaccinated-then-infected cattle. Further development of this assay may provide a useful tool for the diagnosis of FMDV immune animals, including the identification of vaccinated animals that have been subsequently infected with FMDV. In these studies, combining the results of the IFN-gamma assay with virus neutralising antibody titre, in groups of vaccinated animals, provided a correlation with the capacity to control virus replication after subsequent challenge. PMID- 16242819 TI - Redox reaction of iodine in paddy soil investigated by field observation and the I K-Edge XANES fingerprinting method. AB - In order to elucidate the cause for the leaching of iodine in a flooded paddy field, we investigated the transformation of an iodine species affected by the water management of the paddy field. The increased concentration of iodide (I(-)) in soil solution of a flooded paddy field suggested that I(-) was leached from the soil under anaerobic conditions. The post-edge feature of X-ray absorption near-edge structure (XANES) for iodate (IO(3)(-)) spiked to soil totally disappeared after anaerobic incubation of the soils, and I(-) was dissolved in the solution. On the other hand, I(-) in contact with the soil was not likely to be oxidized to IO(3)(-) under aerobic incubation. Iodine was leached out in soil solution as I(-) under anaerobic conditions, whereas part of the iodine species was retained by soil as I(2) or organoiodine both under anaerobic and aerobic conditions. PMID- 16242820 TI - The EXPURT model for calculating external gamma doses from deposited material in inhabited areas. AB - EXPURT, NRPB's model for calculating external gamma doses in inhabited areas, was originally developed in the mid-1980s. Deposition on surfaces in the area, the subsequent transfer of material between different surfaces or its removal from the system, and dose rates in various locations from material on the different surfaces are modelled. The model has been updated to take account of more recent experimental data on the transfer rates between surfaces and to make it more flexible for use in assessing dose rates following an accidental release. EXPURT is a compartmental model and models the transfer of material between the surfaces using a set of first order differential equations. It enables the impact of the decontamination of surfaces on doses and dose rates to be explored. The paper describes the EXPURT model and presents some preliminary results obtained using it. PMID- 16242821 TI - Intramuscular haemangioma within depressor anguli oris. PMID- 16242822 TI - The future of British surgery. PMID- 16242823 TI - Neuroimaging studies of obsessive-compulsive disorder in adults and children. AB - Obsessive-compulsive disorder (OCD) is a severe, highly prevalent and chronically disabling disorder that usually emerges during childhood or adolescence. Neuroimaging studies play an important role in advancing our understanding of the pathophysiology of OCD and in developing neurocircuitry models of this psychiatric illness. This paper provided an updated, comprehensive review and analysis of the relevant literature on baseline functional and structural neuroimaging studies of OCD in both paediatric and adult patients. The neuroanatomical findings were presented in the context of two models: executive dysfunction, which implicates the dorsolateral prefrontal cortex, caudate nucleus, thalamus, and striatum; and modulatory control, which implicates the orbitofrontal and medial prefrontal cortex and the cingulate gyrus. Neuroanatomical findings were not consistent across all studies, and limitations were examined. Recommendations for future research directions and the implications of the results for improved treatment were explored. PMID- 16242824 TI - Correlations of siblings' and mothers' utilisation of primary and hospital health care: a record linkage study in Western Australia. AB - A relationship between maternal and child use of general practitioners (GPs) has been shown to exist for some time, however, the reasons for this relationship are not clear and the extent to which this relationship extends to tertiary care is unknown. The aim of this study was to examine the relationships between the utilisation of health care by siblings and mothers over a 14 year period. A retrospective cohort study of 756 mothers and their 1494 children up to age 14 years was conducted in three general practices in Western Australia. Medicare claims and hospital morbidity records for 1984-1997 were linked using deterministic and probabilistic matching. Generalised Estimating Equations and correlations were used to examine the relationships between the utilisation of primary and hospital health care by family members. Significant correlations were found between hospital admissions of all participants and their GP visits, specialist visits, pathology and diagnostic imaging combined and hospital length of stay. There was a strong association between siblings' use of GPs. A child's rate of GP attendance increased with that of its mother. There was a weak but significant relationship between siblings' use of hospitals, and a child's hospital admission rate increased with that of its mother. It is concluded that there is a strong relationship between siblings' use of GPs and a weaker but still significant association between the hospital admissions of siblings. As expected, there were strong associations between mother and child visits to GPs. There was also an association between a mother's use of hospital and that of her children. This finding reduces the plausibility that the relationships found between utilisation of health care by siblings and mothers can be explained entirely by behavioural factors, and suggests the presence of intergenerational correlation of morbidity. PMID- 16242825 TI - Shifting dollars, saving lives: what might happen to mortality rates, and socio economic inequalities in mortality rates, if income was redistributed? AB - Personal or household income predicts mortality risk, with each additional dollar of income conferring a slightly smaller decrease in the mortality risk. Regardless of whether levels of income inequality in a society impact on mortality rates over and above this individual-level association (i.e., the 'income inequality hypothesis'), the current consensus is that narrowing income distributions will probably improve overall health status and reduce socio economic inequalities in health. Our objective was to quantify this impact in a national population using 1.3 million 25-59-year-old respondents to the New Zealand 1996 census followed-up for mortality over 3 years. We modelled 10-40% shifts of everyone's income to the mean income (equivalent to 10-40% reductions in the Gini coefficient). The strength of the income-mortality association was modelled using rate ratios from Poisson regression of mortality on the logarithm of equivalised household income, adjusted for confounders of age, marital status, education, car access, and neighbourhood socio-economic deprivation. Overall mortality reduced by 4-13% following 10-40% shifts in everyone's income, respectively. Inequalities in mortality reduced by 12-38% following 10-40% shifts in everyone's income. Sensitivity analyses suggested that halving the strength of the income-mortality association (i.e., assuming our multivariable estimate still overestimated the causal income-mortality association) would result in 2-6% reductions in overall mortality and 6-19% reductions in inequalities in mortality in this New Zealand setting. Many commentators have noted the non-linear association of income with mortality predicts that narrowing the income distribution will both reduce overall mortality rates and reduce inequalities in mortality. Quantifying such reductions can only be done with considerable uncertainty. Nevertheless, we tentatively suggest that the gains in overall mortality will be modest (although still potentially worthwhile from a policy perspective) and the reductions in inequalities in mortality will be more substantial. PMID- 16242826 TI - A double-blind sham controlled study of right prefrontal repetitive transcranial magnetic stimulation (rTMS): therapeutic and cognitive effect in medication free unipolar depression during 4 weeks. AB - BACKGROUND: Transcranial magnetic stimulation (TMS) has become a therapeutic tool in psychiatric diseases. METHODOLOGY: The objective was to evaluate the efficacy of TMS in unipolar depression: the percentage of responders (>50% HDRS reduction) and remission (HDRS score < or =8, after four weeks of active TMS treatment in depressed patients free of any antidepressive agent versus placebo-TMS. RESULTS: 27 patients were randomized in two groups: rTMS (N=11) versus sham TMS (N=16). Statistical differences were detected between sham and TMS treated groups on remission (0/16 versus 4/11 p=0.032, 1/16 versus 6/11 0.028 and 1/16 versus 7/11 p=0.011 at day 14, day 21 and day 28, respectively) and on response (2/16 versus 5/11 at day 14 (NS), 2/16 versus 7/11 p=0.0115 at day 21 and 1/16 versus 7/11 (p=0.025) day 28, respectively, using the exact Fisher test). Significant differences were observed between day 1 versus day 8 (p<0.01), day 15, day 21 and day 28 (p<0.001) in TMS group and only versus day 21 (p<0.01) and day 28 (p<0.05) for the sham group. ANOVA comparison between TMS and sham groups was significant at day 14 and day 28 (p<0.05). LIMITATIONS: The few number of patients. CONCLUSION: Our study has shown an efficacy of right rTMS in free medication unipolar depression over a month. Nevertheless, number of patients included is limited and multicentric studies will be necessary to specify the antidepressive action of TMS. PMID- 16242827 TI - Effects of mu-CPP and mesulergine on dietary choices in deprived rats: possible mechanisms of their action. AB - Although it has been well established that compounds that stimulate 5-HT(2C) and/or 5-HT(1B) receptors induce hypophagia by promoting satiety process, the relative role of these receptor subtypes in dietary choices remains to be fully determined. m-CPP is considered a useful probe of 5-HT(2C) receptor function in vivo and its administration reduces food intake and appetite in humans and rats. Conversely, the non-selective 5-HT(2C) receptor antagonist mesulergine elicits feeding in rats. Food intake and dietary choices were measured in a food deprivation experimental protocol employing male Wistar rats. Animals were given access for a 4-h period to a pair of isocaloric diets. These two diets were enriched in protein or carbohydrate proportions, respectively, but fat content was held constant. The mixed 5-HT(2C/1B) receptor agonist, m-CPP, led to a dose dependent hypophagia, due to substantial reduction in carbohydrate consumption while protein intake was spared (0.62, 1.25 and 2.50 mg/kg i.p., respectively). The non-selective 5-HT(2C) receptor antagonist and also D2 agonist, mesulergine, on its own produced a significant dose-dependent increase in both protein and carbohydrate diets (1.0 and 3.0 mg/kg i.p., respectively). Combined treatment with m-CPP, at its maximum effective dose, and mesulergine dose-dependently reversed m-CPP-induced hypophagia, during the 4-h test period. In order to clarify the effects of mesulergine on dietary choices since it is simultaneously a dopamine agonist besides its antiserotonergic properties, the D2 agonist apomorphine was also used. Apomorphine caused a dose-dependent increase in protein intake while carbohydrate and total food intake remained nearly unchanged (0.5 and 1.0 mg/kg i.p., respectively). It is concluded that the mesulergine induced hyperphagic response on both diets is the expression of a dual mode of action, due to its 5-HT(2C) antagonist activity together with D2 agonist properties. The results further indicate that the activation of hypothalamic 5 HT(2C) receptors may be involved in both protein sparing and carbohydrate suppressing effects of 5-HT (m-CPP-like effect), whereas an important role in increase of protein consumption seems to have the dopaminergic system probably through D2 receptors (apomorphine-like and mesulergine-like effects, respectively). PMID- 16242828 TI - Cholecystokinin and endogenous opioid peptides: interactive influence on pain, cognition, and emotion. AB - It is well documented that stressful life experiences contribute to the etiology of human mood disorders. Cholecystokinin (CCK) is a neuropeptide found in high concentrations throughout the central nervous system, where it is involved in numerous physiological functions. A role for CCK in the induction and persistence of anxiety and major depression appears to be conspicuous. While increased CCK has been associated with motivational loss, anxiety and panic attacks, an increase in mesocorticolimbic opioid availability has been associated with coping and mood elevation. The close neuroanatomical distribution of CCK with opioid peptides in the limbic system suggests that there may be an opioid-CCK link in the modulation and expression of anxiety or stressor-related behaviors. In effect, while CCK induces relatively protracted behavioral disturbances in both animal and human subjects following stressor applications, opioid receptor activation may change the course of psychopathology. The antagonistic interaction of CCK and opioid peptides is evident in psychological disturbances as well as stress-induced analgesia. There appears to be an intricate balance between the memory-enhancing and anxiety-provoking effects of CCK on one hand, and the amnesic and anxiolytic effects of opioid peptides on the other hand. Potential anxiogenic and mnemonic influences of site-specific mesocorticolimbic CCK and opioid peptide availability, the relative contributions of specific CCK and opioid receptors, as well as the time course underlying neuronal substrates of long-term behavioral disturbances as a result of stressor manipulations, are discussed. PMID- 16242829 TI - [Direct excision of the deep nasolabial fold: a logical and easy surgical management]. AB - OBJECTIVE: Surgical management of the deep nasolabial fold, due to age, genetic diseases or facial lipoatrophy, is difficult and often short-lived. Direct excision seems to improve the long-term results. Clinical illustration and review of literature about the anatomy and physiology of the nasolabial fold are proposed to explain this fact. MATERIALS AND METHODS: Direct excision of the nasolabial fold has been performed on nine patients, three females and six males. The problem was aging midface in two cases, facial lipoatrophy in five cases, cutis laxa in one case and pachydermoperiostosis in one case. The excision was sitting astride the nasolabial fold and the suture preserved a dog-ear at the upper extremity to restore the projection of the cheek. Evaluation items were quality of the scar, durability of the result and personal improvement. RESULTS: Scars were always unnoticeable within six months, projection of the cheek was restored, and results were durable with an average follow-up of eighteen months. All patients were satisfied with the aesthetic or social improvement. CONCLUSION: Direct excision of the nasolabial fold is a simple, reliable and durable method, respecting the physiology of this area of the face. Then, it should be indicated in deepened nasolabial fold, whatever the etiology. PMID- 16242831 TI - Succinate and artificial maintenance of normal body temperature synergistically correct lethal disorders in thiopental coma rat. AB - Under modeling of thiopental coma influence of sodium succinate and (or) external warming for the support of normal body temperature (isothermal regimen) on the gas exchange, blood gas content, acid-base status and survival rate was studied in rats. In the absence of therapy hypothermia was developed (-9.4 degrees C), O(2) consumption decreased by a factor 5, oxygenation of arterial blood (pO(2)) did not change while that of venous blood increased, where with arteriovenous oxygen tension gradient decreased by half. Blood tension of carbon dioxide (pCO(2)) increased twice, respiratory and metabolic acidosis was developed. Survival rate under absence of a therapy was 42%, with isolated use of isothermal regimen or succinate therapy alike-50%; with their use in combination drastically increased up to 92%. Succinate increased arteriovenous gradient of pO(2), decreased deficit of buffer bases, increased bicarbonate concentration. At isothermal regimen accumulation of CO(2) in the blood was diminished, its excretion was increased, pH of blood approached normal values. Combined use of both therapy agents increased O(2) consumption and potentiated their positive influence on acid-base status. The implication is that hypothermia restrains effect of succinate in barbiturate coma; prevention of hypothermia in combination with succinate administration is highly effective method of experimental therapy of barbiturate intoxication. PMID- 16242830 TI - Influence of a low-molecular-weight heparin derivative on the nitric oxide levels and apoptotic DNA damage in adriamycin-induced cardiac and renal toxicity. AB - The spectrum of the anti-apoptotic potential of heparin is currently under scrutiny in various tissues and under various pathological situations. In this study, the role of a low-molecular-weight heparin derivative (LMWH), certoparin in adriamycin-induced oxidative DNA damage has been evaluated in the cardiac and renal tissues. Two groups of male albino rats of the Wistar strain (140+/-10 g) received a single intravenous injection of adriamycin (7.5mg/kg), and one of them received low-molecular-weight heparin (Certoparin Sodium, 300 microg/day/rat s.c.) treatment, commencing on day 8, continued for a week. The nitrosative stress in ADR cytotoxicity is indicated by the 1.51-fold cardiac and 2.36-fold renal increase in reactive nitrogen species (RNS), while LMWH treatment restores normalcy (p<0.001). The influence of LMWH on the pro-inflammatory and pro apoptotic cytokine, TNF-alpha was studied. Renal and cardiac levels of TNF-alpha showed a significant rise (p<0.001) in the ADR cytotoxic group, while the TNF alpha values departed towards control levels in the LMWH treated group (p<0.001). DNA damage indicated by the fragmentation pattern (agarose gel electrophoresis) and the significantly increased comet tail length (p<0.001) observed after alkaline single cell gel electrophoresis confirmed the toxicity induced by ADR on DNA in the untreated group. In the LMWH-treated group, the observation of intact DNA band after agarose gel electrophoresis, and the finding of comet tail length being comparable with that of the control substantiated the protection rendered by the LMWH, certoparin. In short, the results suggest that the low-molecular weight heparin derivative, certoparin exerts beneficial effects on the nitrosative status, and on the biological macromolecules as DNA and curtails the rise of the pro-apoptotic and pro-inflammatory cytokine, TNF-alpha in the cardiac and renal tissues. PMID- 16242832 TI - Induction and expression of GST-Pi foci in the liver of Cyclophosphamide administered rats. AB - Anticancer drug Cyclophosphamide (CY) is metabolized to phosphoramide mustard and acrolein by the hepatic P450 enzymes. GST-Pi is a biochemical feature which occurs in carcinogen induced preneoplastic foci and it plays an important role in the detoxification pathway of acrolein metabolism. Administration of CY induces GST-Pi positive single cells and foci expression in rat liver and these can be considered as precursors of preneoplastic foci leading to hepatocarcinogenesis. The expression of GST-Pi in CY-treated rats on different days of treatment was confirmed by immunohistochemistry, immunoblot, RT-PCR and by ELISA. We also advocate that epigenetic mechanism could be accounted for the GST-Pi induction in the hepatocytes of CY-treated rats. PMID- 16242833 TI - Use of low-molecular-weight heparin from the first trimester of pregnancy: a retrospective study of 111 consecutive pregnancies. AB - BACKGROUND: During the first trimester of pregnancy, unfractionated heparin is the standard anticoagulant treatment for pregnant women at high risk of thrombosis. OBJECTIVE: To observe maternal and fetal tolerance for low-molecular weight heparin begun in the first trimester of pregnancy. METHODS: Observational study conducted from 1 January 1997 to 31 May 2001. All patients began treatment before the 15th week of pregnancy. The outcome measures were the incidence and causality of adverse events in mother and fetus. RESULTS: The study included 97 patients (and 111 pregnancies) at very high risk for thrombosis. Seven fetal losses (6.3%) were observed: three early spontaneous abortions, three late spontaneous abortions and one medically indicated abortion. Twenty-five (22.5%) bleeding events occurred during pregnancy, seven (6.3%) of which required medical intervention: five curettages for first trimester spontaneous abortions, one late abortion at 21 weeks and one placental abruption at 25 weeks. Of nine (8.1%) primary postpartum hemorrhages involving a blood loss > or = 500 mL, three involved losses of 1000 mL or more and one required embolization of the uterine arteries. Five patients had thrombocytopenia, but none was treatment-related. Local cutaneous reactions occurred in 33 (29.7%) patients. Six (5.4%) maternal thromboembolic complications occurred during pregnancy or postpartum. At birth, two children had non-chromosomal congenital malformations (pyelectasia, cleft lip and palate). No fetal or neonatal complication was attributed to the treatment. CONCLUSION: The use of low-molecular-weight heparin (LMWH) for patients requiring anticoagulant treatment from the first trimester appears safe for mother and fetus. PMID- 16242834 TI - Deuteration can affect the conformational behaviour of amphiphilic alpha-helical structures. AB - The replacement of hydrogen with deuterium is frequently used in conjunction with neutron diffraction to investigate peptide-membrane interaction. This isotopic substitution in an amino acid residue radically changes the neutron scatter pattern of the peptide, thereby allowing its localisation within the bilayer with the aid of derived Fourier maps. Nonetheless, this technique relies on the generally held assumption that normal and isotopically enriched protein species do not differ significantly in structure or biological activity. Recently, this assumption has been questioned and here, diffraction data from studies on a membrane interactive peptide clearly challenge the reliability of this assumption. PMID- 16242835 TI - Effects of lactose permease of Escherichia coli on the anisotropy and electrostatic surface potential of liposomes. AB - The membrane transport protein lactose permease (LacY), a member of the Major Facilitator Superfamily (MFS) containing twelve membrane-spanning segments connected by hydrophilic loops, was reconstituted in liposomes of: (i) 1,2 dimyristoyl-sn-glycero-3-phosphocoline (DMPC) and 1-palmitoyl-2-oleoyl-sn-glycero 3-phosphocholine (POPC) in equimolar proportions; and (ii) Escherichia coli total lipid extract. The structural order of the lipid membranes, in the presence and absence of LacY, was investigated using steady-state fluorescence anisotropy. The features of the anisotropy curves obtained with 1,6-phenyl-1,3,5-hexatriene (DPH) and 1-(4-trimethylammoniumphenyl)-6-phenyl-1,3,5-hexatriene p-toluene sulfonate (TMA-DPH) evidenced: (i) the insertion of LacY into the bilayer; and (ii) a surface effect on the membranes. The most dramatic effects were observed when LacY was reconstituted in the E. coli lipid matrix. The effect of the protein on the electrostatic surface potential of each bilayer was also examined using a fluorescent pH indicator, 4-Heptadecyl-7-hydroxycoumarin (HHC). Changes in surface potential were enhanced in the presence of the substrate (i.e. lactose) only when the lipid matrices were charged. These results suggest a role for charged phospholipids (i.e. phosphatidylethanolamine or phosphatidylglycerol) in proton transfer to the amino acids involved in substrate translocation. PMID- 16242836 TI - How thick is the layer of thermal volume surrounding the protein? AB - Investigation on the volume properties of protein hydration layers is reported. Presented results are based on combination of Monte Carlo modeling and available experimental data. Six globular proteins with known data are chosen for analysis. Analyzing the model and the experimental results we found that water molecules bound to proteins by hydrogen bond are preferentially located at the places with local depressions on the protein surface. Consequently, the hydration level is not strictly proportional to the area of charged and polar surfaces, but also depends on the shape of the molecular surface. The thickness of the thermal volume layer as calculated in the framework of the scaled particle theory is 0.6 0.65 A for chosen proteins. The obtained value is significantly lower than that presented for proteins in earlier papers (where proportionality between the hydration level and the area of charged and polar surfaces was assumed), but is close to the value published for small solute molecules. Discussion including the influence of protein size and the thermal motion of the surface is presented. PMID- 16242837 TI - 5th International Conference on Systems Biology (ICSB 2004), Heidelberg, October 9-13, 2004. PMID- 16242838 TI - Oncogenic rearrangements of the NTRK1/NGF receptor. AB - The NTRK1 gene encodes the high affinity receptor for Nerve Growth Factor, and its action regulates neural development and differentiation. Deregulation of NTRK1 activity is associated with several human disorders. Loss of function mutations causes the genetic disease congenital insensitivity to pain with anhidrosis (CIPA). Constitutive activation of NTRK1 has been detected in several tumor types. An autocrine loop involving NTRK1 and NGF is associated with tumor progression in prostate carcinoma and in breast cancer. A novel alternative splicing variant with constitutive oncogenic potential has been recently described in neuroblastoma. Somatic rearrangements of NTRK1, producing chimeric oncogenes with constitutive tyrosine kinase activity, have been detected in a consistent fraction of papillary thyroid tumors. The topic of this review is a detailed analysis of the thyroid TRK oncogenes. The modalities of their activation, their mechanism of action, the contribution of activating sequences, and the molecular mechanisms underlying their generation will be discussed. PMID- 16242839 TI - Mutations in the mitochondrial DNA D-Loop region occur frequently in human osteosarcoma. AB - To explore the polymorphisms and mutations of mitochondrial D-Loop region in osteosarcoma and their possible association with carcinogenesis, direct DNA sequencing method was used to detect the variants of the mitochondrial D-Loop in 20 patients with osteosarcoma. We found mutations of the mitochondrial D-Loop in 14/20 (70%) of the tested osteosarcoma samples, including 34 novel mutations. We did not detect any new polymorphisms in osteosarcomas, nor was there any association between variants and the three histopathological subtypes. We identified 89 variant sites, most of which were within the hypervariable I and II (HV I and HV II) regions. These data suggest that mtDNA mutations within the D Loop region, particularly the HV I and HV II segments, are a frequent event in osteosarcomas. PMID- 16242840 TI - Common chromosomal fragile sites and cancer: focus on FRA16D. AB - A growing body of experimental evidence supports the view that certain human chromosomal fragile sites have roles to play in cancer. The principle lines of evidence are at the level of mutation mechanism and gene function. Most research in this area has previously focussed on the FRA3B common fragile site and the FHIT gene that spans this site. Here we review recent progress in characterising the second most readily observed common fragile site, FRA16D, and the WWOX gene that spans it. Comparative analyses of FRA3B/FHIT and FRA16D/WWOX reveal some striking similarities suggesting that these sites and their associated genes may play a part in a normal protective response of cells to environmental stress. PMID- 16242841 TI - Adsorption of landfill leachates onto activated carbon. Equilibrium and kinetics. AB - The adsorption of stabilized leachates generated in a municipal landfill onto three commercial activated carbons has been investigated. Norit 0.8, Chemviron AQ40 and Picacarb 1240 have been used as adsorbents. Equilibrium experiments have been conducted to obtain the experimental isotherm profiles. Isotherms have been plotted based on the adsorption of general parameters, for instance chemical oxygen demand, total carbon, absorption at 410 nm and absorption at 254 nm. Different literature models and error functions have been used to adequately fit the experimental data. As a rule of thumb, three-parameter models do adjust experimental results better than two-parameter models. Norit 0.8 shows better adsorption characteristics than the rest of activated carbons, both in terms of contaminant level reduction of per unit mass of absorbent and in terms of the process kinetics. PMID- 16242842 TI - Utilization of MSWI fly ash for stabilization/solidification of industrial waste sludge. AB - This work investigated the potential for utilization of MSWI incineration fly ash as solidification binder to treat heavy metals-bearing industrial waste sludge. In the study, Municipal Solid Waste Incineration (MSWI) fly ash was used along with ordinary Portland cement to immobilize three different types of industrial sludge while MSWI incineration fly ash was stabilized at the same time. The results showed that the matrixes with heavy metals-bearing sludge and MSWI fly ash have a strong fixing capacity for heavy metals: Zn, Pb, Cu, Ni and Mn. Specimens with only 5-15% cement content was observed to be sufficient to achieve the target compressive strength of 0.3 MPa required for landfill disposal. An optimum mix comprising 45% fly ash, 5% cement and 50% of the industrial sludge could provide the required solidification and stabilization. Addition of MSWI can improve the strength of matrix. Meanwhile, the main hydration products of new S/S matrix are ettringite AFt, Friedel's salt and C-S-H. These hydration products play an important role in the fixing of heavy metals. The co-disposal of MSWI fly ash with heavy metals-bearing sludge can minimize the enlargement of the landfill volume and stabilize the heavy metals effectively. PMID- 16242843 TI - Transfer mechanisms of contaminants in cement-based stabilized/solidified wastes. AB - Stabilization/solidification (S/S) processes are routinely used for the final treatment of hazardous wastes prior to land disposal. Cement-based binder systems with partial replacement of cement by pulverized fuel ash (PFA) are common. In order to predict the long-term leaching characteristics of S/S wastes, it is important to understand the leaching mechanism. This paper presents experimental results from a study that has investigated the leaching behaviour of contaminants from cement-based S/S waste forms. A novel radial leach test (RLT) has been used to study the migration of heavy metals. The results show that contaminants migrate from the inner core of the S/S waste during leaching to the sample surface and accumulate near the edge of the S/S waste. The degree of accumulation is related to the contaminant species and the Ca(OH)2 content. PMID- 16242844 TI - Canine visceral leishmaniasis: comparison of in vitro leishmanicidal activity of marbofloxacin, meglumine antimoniate and sodium stibogluconate. AB - The control of canine leishmaniasis largely depends on the success of treatment. Drugs currently available to treat this disease are toxic and partially effective. The curative effect of marbofloxacin, a third-generation fluoroquinolone developed for veterinarian individual treatment, was evaluated in vitro in the presence of Leishmania infantum promastigotes and dog-monocyte derived macrophages; meglumine antimoniate and sodium stibogluconate were used as comparative treatments. We observed that the killing of Leishmania promastigotes and intracellular amastigotes by marbofloxacin was dose-dependent. We demonstrated that successful treatment of canine infected macrophages for 48 h was possible with 500 microg/ml of marbofloxacin. Leishmanicidal activity acted through a TNF-alpha and nitric oxide pathway and correlated with the generation of nitric oxide (NO(2)) production by monocytes derived macrophages from infected (23+/-5 microM) or healthy (21+/-6 microM) dogs, in comparison with NO(2) concentration in infected/non-treated macrophages (< 3 microM, P<0.01). This significant induced parasiticidal effect correlated with extensive elimination of amastigotes by macrophages derived from infected (11+/-5) and healthy dogs (6+/ 2), when compared to infected/non-treated macrophages (530+/-105 and 472+/-86 amastigotes, respectively, P< 0.01). Marbofloxacin was shown to be non-toxic at 500 microg/ml in vitro and no cell apoptosis was observed. The molecule was able to induce a parasitic process after significant elimination of amastigotes in leishmania-infected dog macrophages. We propose that marbofloxacin, compared to standard chemotherapeutic agents (meglumine antimoniate and sodium stibogluconate), could be an effective and pragmatic oral route alternative to treat canine leishmaniasis. PMID- 16242845 TI - cDNA cloning and functional expression of KM+, the mannose-binding lectin from Artocarpus integrifolia seeds. AB - KM+, a mannose-binding lectin present in the seeds of Artocarpus integrifolia, has interesting biological properties and potential pharmaceutical use [A. Panunto-Castelo, M.A. Souza, M.C. Roque-Barreira, J.S. Silva, KM(+), a lectin from Artocarpus integrifolia, induces IL-12 p40 production by macrophages and switches from type 2 to type 1 cell-mediated immunity against Leishmania major antigens, resulting in BALB/c mice resistance to infection, Glycobiology 11 (2001) 1035-1042. ; L.L.P. daSilva, A. Panunto-Castelo, M.H.S. Goldman, M.C. Roque-Barreira, R.S. Oliveira, M.D. Baruffi, J.B. Molfetta-Machado, Composition for preventing or treating appearance of epithelia wounds such as skin and corneal wounds or for immunomodulating, comprises lectin, Patent number WO20041008.]. Here, we have isolated clones encoding the full-length KM+ primary sequence from a cDNA library, through matrix PCR-based screening methodology. Analysis of KM+ nucleotide and deduced amino acid sequences provided strong evidence that it neither enters the secretory pathway nor undergoes post translational modifications, which is in sharp contrast with jacalin, the more abundant lectin from A. integrifolia seeds. Current investigations into the KM+ properties are often impaired by the difficulty in obtaining sufficient quantities of jacalin-free KM+ through direct seed extraction. To obtain active recombinant protein (rKM+) in larger amounts, we tested three different expression systems. Expression vectors were constructed to produce: (a) rKM+ in E. coli in its native form, (b) rKM+ with GST as an N-terminal tag and (c) native rKM+ in Saccharomyces cerevisiae. The presence of the GST-tag significantly improved the overall rKM+ yield; however, most of the obtained rGST-KM+ was insoluble. Production of rKM+ in the yeast host yielded the highest quantities of soluble lectin that retained the typical high-mannose oligosaccharide-binding properties of the natural protein. The possible biotechnological applications of recombinant KM+ are discussed. PMID- 16242846 TI - Experimental models of traumatic brain injury: do we really need to build a better mousetrap? AB - Approximately 4000 human beings experience a traumatic brain injury each day in the United States ranging in severity from mild to fatal. Improvements in initial management, surgical treatment, and neurointensive care have resulted in a better prognosis for traumatic brain injury patients but, to date, there is no available pharmaceutical treatment with proven efficacy, and prevention is the major protective strategy. Many patients are left with disabling changes in cognition, motor function, and personality. Over the past two decades, a number of experimental laboratories have attempted to develop novel and innovative ways to replicate, in animal models, the different aspects of this heterogenous clinical paradigm to better understand and treat patients after traumatic brain injury. Although several clinically-relevant but different experimental models have been developed to reproduce specific characteristics of human traumatic brain injury, its heterogeneity does not allow one single model to reproduce the entire spectrum of events that may occur. The use of these models has resulted in an increased understanding of the pathophysiology of traumatic brain injury, including changes in molecular and cellular pathways and neurobehavioral outcomes. This review provides an up-to-date and critical analysis of the existing models of traumatic brain injury with a view toward guiding and improving future research endeavors. PMID- 16242847 TI - Prenatal stress and neonatal rat brain development. AB - Chronic or repeated stress during human fetal brain development has been associated with various learning, behavioral, and/or mood disorders, including depression in later life. The mechanisms accounting for these effects of prenatal stress are not fully understood. The aim of this study was to investigate the effects of prenatal stress on early postnatal brain development, a disturbance of which may contribute to this increased vulnerability to psychopathology. We studied the effects of prenatal stress on fetal growth, stress-induced corticosterone secretion, brain cell proliferation, caspase-3-like activity and brain-derived neurotrophic factor protein content in newborn Fischer 344 rats. In addition to a slight reduction in birth weight, prenatal stress was associated with elevated corticosterone levels (33.8%) after 1 h of maternal deprivation on postnatal day 1, whereas by postnatal day 8 this pattern was reversed (-46.5%). Further, prenatal stress resulted in an approximately 50% decrease in brain cell proliferation just after birth in both genders with a concomitant increase in caspase-3-like activity within the hippocampus at postnatal day 1 (36.1%) and at postnatal day 5 (females only; 20.1%). Finally, brain-derived neurotrophic factor protein content was reduced in both the olfactory bulbs (-24.6%) and hippocampus (-28.2%) of prenatally stressed male offspring at postnatal days 1 and 5, respectively. These detrimental central changes observed may partly explain the increased susceptibility of prenatally stressed subjects to mood disorders including depression in later life. PMID- 16242848 TI - The metabotropic glutamate receptor subtype 5 antagonist MPEP and the Na+ channel blocker riluzole show different neuroprotective profiles in reversing behavioral deficits induced by excitotoxic prefrontal cortex lesions. AB - Overactivation of excitatory amino acid receptors has been involved in several neurodegenerative diseases. The present study aims at investigating the potential neuroprotective action of 2-methyl-6-(phenylethylnyl)-pyridine (MPEP), a selective non-competitive antagonist of metabotropic glutamate receptor subtype 5, and 2-amino-6-trifluoro methoxy-benzothiole (riluzole), a Na+ channel blocker exhibiting anti-glutamatergic properties, on the ibotenate-induced damage to the rat medial prefrontal cortex. The neuroprotective efficacy of these compounds was assessed on the recovery from behavioral deficits induced by prefrontal cortical excitotoxic lesions in a reaction time task. MPEP (3, 10 or 30 mg/kg) or riluzole (2, 4 or 8 mg/kg) was administered i.p. 30 min before and after medial prefrontal cortex lesions. As previously found, lesions to the medial prefrontal cortex significantly altered the motor preparatory processes involved in the reaction time task. These deficits were prevented by MPEP 3 mg/kg and riluzole 2 mg/kg while higher doses of either compound were ineffective. Furthermore, the neuron specific nuclear protein immunostaining of the lesioned cortical area in animals treated with the efficient dose of either compound revealed that MPEP reduced the volume of the lesion whereas riluzole reversed the decrease of neuronal density within the lesioned area. Altogether, these results suggest a neuroprotective action of MPEP as well as riluzole at both behavioral and cellular levels on excitatory amino acid-induced toxicity. PMID- 16242850 TI - When a line is a number: color yields magnitude information in a digit-color synesthete. AB - The phenomenon of synesthesia has received a great deal of interest recently in the scientific literature. Many previous studies stressed the unidirectional nature of this phenomenon. For example, color-grapheme synesthetes automatically perceive achromatic numbers as colored (e.g. 7 is turquoise). Conversely, colors do not automatically give rise to any sort of number experience (e.g. turquoise is 7). In contrast to the common view, we report on a digit-color synesthete in whom colors can evoke numerical representations in the absence of any digit presentation. It is concluded that in synesthesia there is a reciprocal rather than unidirectional flow of information between dimensions. PMID- 16242849 TI - Galantamine increases excitability of CA1 hippocampal pyramidal neurons. AB - Galantamine is a third generation cholinesterase inhibitor and an allosteric potentiating ligand of nicotinic acetylcholine receptors. It enhances learning in aging rabbits and alleviates cognitive deficits observed in patients with Alzheimer's disease. We examined galantamine's effect on CA1 neurons from hippocampal slices of young and aging rabbits using current-clamp, intracellular recording techniques. Galantamine (10-200 microM) dose-dependently reduced the postburst afterhyperpolarization and the spike-frequency accommodation of CA1 neurons from both young and aging animals. These reductions were partially, but significantly, reversed by the addition of the muscarinic receptor antagonist, atropine (1 microM), to the perfusate. In contrast, the nicotinic acetylcholine receptor antagonist, alpha-bungarotoxin (10 nM), had no effect; i.e. alpha bungarotoxin did not reverse the afterhyperpolarization and accommodation reductions. The allosteric potentiating ligand effect was examined by stimulating the Schaffer collateral and measuring the excitatory postsynaptic potentials for 30 min during bath application of galantamine. Galantamine (200 microM) significantly enhanced the excitatory postsynaptic potential amplitude and area over time. These effects were blocked by 10 nM alpha-bungarotoxin, supporting a role for galantamine as an allosteric potentiating ligand. We did not observe a facilitation of the excitatory postsynaptic potentials with 1 microM galantamine. However, when the excitatory postsynaptic potential was pharmacologically isolated by adding 10 microM gabazine (GABA(A) receptor antagonist) to the perfusate, 1 microM galantamine potentiated the subthreshold excitatory postsynaptic potentials into action potentials. We propose that the learning enhancement observed in aging animals and the alleviation of cognitive deficits associated with Alzheimer's disease after galantamine treatment may in part be due to the enhanced function of both nicotinic and muscarinic excitatory transmission on hippocampal pyramidal neurons. PMID- 16242851 TI - Major depression is not a diagnosis, it is a departure point to differential diagnosis -- clinical and hormonal considerations (a commentary and elaboration on Antonejevic's paper). AB - Currently, the prevailing diagnostic systems of mental disorders, including Depressions and anxieties are based on descriptive phenomenological approach and not on etiology and pathophysiology. However, cumulative knowledge suggests that the field is moving towards establishment of subtypes or phenotypes of depressions based on different pathophysiologies, which may result in differential treatment responses. What is currently defined as 'Major Depressive Disorder (MDD)' may be the end-result of multiple heterogeneous biological and psychosocial processes. Therefore, MDD is actually a first step towards a differential diagnosis that eventually may lead to pathophysiology-based entities. Several hormonal systems may play a major role in the pathophysiology of several subtypes of depressions and anxieties. They include but are not limited to dysregulation of the hypothalamic-pituitary-adrenal (HPA) system, HP Gonadal (HPG) and Thyroid (HPT) systems as well as their interactions with immune and other biological and psychosocial processes. Elucidation of the multiple underlying mechanisms of depressions and anxieties should lead to targeted treatment modalities and improvement of treatment responses. PMID- 16242852 TI - Spinal manipulation and spinal mobilization influence different axial sensory beds. AB - Manipulation and mobilization are two forms of manual therapy commonly employed in the management of musculoskeletal disorders. Spinal manipulation and mobilization are often distinguished from one another by reference to certain biomechanical parameters such as peak force, duration and magnitude of translation. However, as of yet, there is relatively little research which distinguishes between them in terms of neurological mechanisms or clinical effectiveness. Theories concerning the mechanisms underlying the therapeutic effects of manipulation and mobilization commonly make reference to mechanical events such as the release of entrapped tissue or the disruption of intra articular adhesions. Relatively less attention is given to neural effects. In this paper, we hypothesize that, at least in part, spinal manipulation preferentially influences a sensory bed which, in terms of anatomical location and function, is different from the sensory bed influenced by spinal mobilization techniques. More specifically, we hypothesize that manipulation may particularly stimulate receptors within deep intervertebral muscles, while mobilization techniques most likely affect more superficial axial muscles. In part, our rationale for this hypothesis is based on differences in mechanical advantage of the respective manual procedures on multi-segmental versus short intervertebral muscles. PMID- 16242853 TI - Hypoplastic left heart syndrome: Rheumatic heart disease of the fetus? AB - Hypoplastic left heart syndrome (HLHS) accounts for nearly 25% of deaths among neonates with congenital heart disease. The essential feature of HLHS is a small left ventricle (LV) incapable of supporting the circulation. The etiology of HLHS is unknown. A hypothesis is proposed implicating an immune mechanism involving maternal antibodies produced in response to pharyngitis caused by group A beta hemolytic streptococci (GABHS) ("strep throat"). After crossing the placenta, the antibodies injure the developing fetal heart, leading to HLHS either because of direct injury to the LV or secondary to reduced blood flow through affected aortic and mitral valves. Analogy is drawn to rheumatic heart disease (RHD), a known sequela of strep throat. In RHD a misdirected immune response originally intended for GABHS leads to cardiac injury through "molecular mimicry"; the normal heart antigens supposedly mimic the GABHS antigens. A similar pathogenesis is proposed for HLHS and related heart defects. HLHS may represent an extreme form of injury, while a milder insult may present as only mild aortic stenosis or a bicuspid aortic valve, conditions with wide prevalence among the general population. The injury may indeed superimpose on many other congenital heart defects, leading to a variable presentation of these other diseases. Beside remarkable likenesses between HLHS and RHD, the hypothesis is also supported by increasing evidence for the role of deleterious transplacental antibodies in the pathogenesis of other fetal diseases. Implications for other congenital heart diseases and the broader picture of global public health are discussed. PMID- 16242854 TI - Disturbance of microcirculation due to unhealthy lifestyle: Cause of type 2 diabetes. AB - Recently, type 2 diabetes seems to be increasing annually in all developed countries. The outcome of type 2 diabetes is often tragic due to succession of complications including renal disorders requiring hemodialysis, blindness, and limb amputation. The expenses for the care of diabetic patients are also a large burden on the society. These circumstances strongly indicate the necessity of prevention. For satisfactory prevention, the clarification of the etiology related to lifestyle is important, but it remains insufficient to date. In this paper, we present a hypothesis of the etiology of type 2 diabetes from the viewpoint of microcirculation. As mentioned later, an unhealthy lifestyle first causes disturbance of the microcirculation, and a portion of the blood is considered to bypass the capillaries via arteriovenous shunts. This prevents the delivery of glucose and insulin to cells of peripheral tissues, causing hyperglycemia unrelated to the cell insulin sensitivity or the endocrine state, i.e., apparent reduction of insulin sensitivity. Disturbance of the microcirculation also causes oxidative stress in peripheral tissues by inducing ischemia and hypoxia. This oxidative stress is considered to further exacerbate reduction of insulin sensitivity. This hypothesis is supported by the well-known fact that insulin sensitivity recovers with improvement in lifestyle including moderate exercise. PMID- 16242855 TI - A new nortriterpenoid from Rostrinucula dependens. AB - A new nortriterpenoid rostrinucin (1), together with five known compounds were isolated from Rostrinucula dependens. The structure of 1 was elucidated by spectroscopic evidence. PMID- 16242856 TI - Distribution of anticancer drug camptothecin in Nothapodytes foetida. AB - The topoisomerase I-DNA inhibitor alkaloid camptothecin has been evaluated from the various parts of Nothapodytes foetida. The bark contained 0.27% dry wt of camptothecin and 0.11% dry wt 9-methoxycamptothecin followed by the root, stem, and leaves. Immature seeds contained higher concentrations of camptothecin (0.32% dry wt) and 9-methoxycamptothecin (0.16% dry wt) compared to mature seeds. Various parts of mature and immature seeds were analysed to determine the content of the major alkaloids. Zygotic embryos of immature seeds contained 0.11% dry wt of camptothecin and 0.04% dry wt of 9-methoxycamptothecin. The highest concentration of camptothecin (0.42% dry wt) and 9-methoxycamptothecin (0.18% dry wt) were accumulated in the cotyledons of immature seeds. PMID- 16242857 TI - New prenylflavans from Cyperus conglomeratus. AB - In addition to luteolin and its 7-methyl ether, the CH2Cl2-MeOH (1:1) extract of Cyperus conglomeratus afforded two new prenylflavans identified as 7,3'-dihydroxy 5,5'-dimethoxy-8-prenylflavan and 5,7,3'-trihydroxy-5'-methoxy-8-prenylflavan. The structures were established by CIMS, 1H-NMR, 13C-NMR, H-H COSY, HMQC, HMBC and DEPT analysis. PMID- 16242858 TI - Effects of maternal separation, early handling, and standard facility rearing on orienting and impulsive behavior of adolescent rats. AB - Effects of maternal separation in rats have been extensively investigated, but no studies have examined its effects in rat adolescence. We examined the effects of neonatal infant-mother separation (MS) for 6h/day and early handling (EH) for 10 days during the first 2 weeks of life by comparing MS and EH groups to standard facility reared (SFR) controls. At adolescence, the animals were evaluated in a novel and familiar open-field, the light-dark box, and the sucrose consumption test. Behavioral indices included orienting behavior (rearing frequency and duration), impulsive behavior (movement velocity and risk taking by entering the center of the open field or the light compartment of the light-dark box), hyperactivity (ambulatory distance and stereotypic movement), and reward-seeking behavior (sucrose drinking time). The prolonged MS during the first 2 weeks of life resulted in decreased orienting behavior and increased impulsive behavior in adolescence. Measures of ambulatory and stereotypic movements showed that MS rats were hyperactive in the novel environment whereas EH rats were less active overall. The impulsive/hyperactive phenotype produced by this MS protocol may provide a useful animal model to investigate the neurological basis for the similar behavioral phenotype found in attention deficit/hyperactivity disorder. PMID- 16242859 TI - The contribution of latent inhibition to reduced generalization after pre exposure to the test stimulus. AB - Two experiments assessed the contribution of latent inhibition to the generalization-reducing effects of pre-exposure to the test stimulus using a taste aversion procedure in rats. In both experiments, lithium chloride induced illness was paired with a flavor compound (AX) of either salt or sugar (A or B) and hydrochloric acid (X). Generalization of the resulting aversion to a test compound (BX), was assessed after varying pre-exposure to BX, X, and B. Experiment 1 showed that generalization to BX was less when BX itself had been exposed than equivalent pre-exposure to either B and X separately or to B and a new compound (CX). Experiment 2 showed that levels of generalization varied directly as a function of the amount of pre-exposure to BX. The findings show that latent inhibition alone cannot account for the generalization-reducing effect of pre-exposure to BX. PMID- 16242860 TI - Loop-mediated isothermal amplification for the rapid detection of Salmonella. AB - Loop-mediated isothermal amplification (LAMP) assay detected Salmonella within 60 min. The 220 strains of 39 serotypes of Salmonella subsp. enterica and 7 strains of Salmonella enterica subsp. arizonae were amplified, but not 62 strains of 23 bacterial species other than Salmonella. The sensitivity of the LAMP assay was found to be >2.2 cfu/test tube using nine serotypes. The specificity was similar to that of a PCR assay, but the sensitivity of LAMP was greater. Both fluorescence and turbidity were able to detect the products in the LAMP assay. S. enteritidis in a liquid egg sample artificially inoculated with the organism was detected by the LAMP assay at 2.8 cfu/test tube, although negative by PCR assay. These results indicate that the LAMP assay is a rapid, specific and sensitive detection method for Salmonella. PMID- 16242861 TI - Genome-wide expression analysis of iron regulation in Burkholderia pseudomallei and Burkholderia mallei using DNA microarrays. AB - Burkholderia pseudomallei and B. mallei are the causative agents of melioidosis and glanders, respectively. As iron regulation of gene expression is common in bacteria, in the present studies, we have used microarray analysis to examine the effects of growth in different iron concentrations on the regulation of gene expression in B. pseudomallei and B. mallei. Gene expression profiles for these two bacterial species were similar under high and low iron growth conditions irrespective of growth phase. Growth in low iron led to reduced expression of genes encoding most respiratory metabolic systems and proteins of putative function, such as NADH-dehydrogenases, cytochrome oxidases, and ATP-synthases. In contrast, genes encoding siderophore-mediated iron transport, heme-hemin receptors, and a variety of metabolic enzymes for alternative metabolism were induced under low iron conditions. The overall gene expression profiles suggest that B. pseudomallei and B. mallei are able to adapt to the iron-restricted conditions in the host environment by up-regulating an iron-acquisition system and by using alternative metabolic pathways for energy production. The observations relative to the induction of specific metabolic enzymes during bacterial growth under low iron conditions warrants further experimentation. PMID- 16242862 TI - The Clostridium botulinum GerAB germination protein is located in the inner membrane of spores. AB - Clostridium botulinum dormant spores germinate in presence of l-alanine via a specific receptor composed of GerAA, GerAB and GerAC proteins. In Bacillus subtilis spores, GerAA and GerAC proteins were located in the inner membrane of the spore. We studied the location of the GerAB protein in C. botulinum spore fractions by Western-blot analysis, using an antipeptidic antibody. The protein GerAB was in vitro translated and used to confirm the specificity of the antibodies. GerAB was not present in a coat and spore outer membrane fraction but was present in a fraction of decoated spores containing inner membrane. These results strongly suggest that the protein GerAB is located in the inner membrane of the spore. PMID- 16242863 TI - The cell surface of Lactobacillus reuteri ATCC 55730 highlighted by identification of 126 extracellular proteins from the genome sequence. AB - Bioinformatical analyses of a draft genome sequence of the commensal bacterium Lactobacillus reuteri ATCC 55730 revealed 126 genes encoding putative extracellular proteins. The function, localization and distribution in bacterial species were predicted. Interestingly, few proteins possessed LPXTG motifs or C terminal transmembrane anchors. Instead eight proteins were putatively anchored by GW repeats and several secreted proteins were likely to be re-associated to the surface. The majority of the extracellular proteins were widely distributed, i.e., found universally or in gram-positive bacteria, but 24 were only detected in L. reuteri. Further, the number of transporters was lower, while the number of enzyme was higher than in related species. PMID- 16242864 TI - Functional coupling between vanillate-O-demethylase and formaldehyde detoxification pathway. AB - Pseudomonas putida vanillate-O-demethylase consisting of VanA and VanB was expressed in Escherichia coli strain K-12. Recombinant E. coli strain K-12 cells expressing VanAB efficiently converted vanillate into protocatechuate with glucose consumption. Mutant lacking either pgi or zwf showed higher or lower converting activity than the parental strain, respectively. Formaldehyde, which is the by-product of the demethylation, was converted into formate in the cellular reaction. Formate accumulation was blocked by gene disruption of the E. coli frmA that coded glutathione-dependent formaldehyde dehydrogenase. PMID- 16242865 TI - Functional divergence of duplicated c-myc genes in a tetraploid fish, the common carp (Cyprinus carpio). AB - The proto-oncogene c-myc is thought to be one of the most important genes in controlling cell proliferation. In a tetraploid fish, two c-myc genes (CAM1 and CAM2) were previously isolated from the common carp, Cyprinus carpio, and were shown to have different expression patterns in adult tissues. Here we found that CAM1 and CAM2 proteins had distinct properties in terms of their transcription regulation system, formation of the transcription activator complex Myc/Max, and transcriptional activation of the target gene. These results showed that the two carp c-Myc proteins have overlapping but distinct functions, suggesting that CAM1 and CAM2 are evolving to acquire different functions after an earlier tetraploidization event. PMID- 16242866 TI - Specific adhesion and invasion of Salmonella Enteritidis in the vagina of laying hens. AB - Salmonella Enteritidis is the predominant serovar associated with egg-borne salmonellosis in humans. The colonization of S. Enteritidis in the vagina may play a role in the production of S. Enteritidis-contaminated eggs. In the first experiment, the in vitro adhesion of S. Enteritidis in vaginal and follicular explants was compared with that of S. Typhimurium by bacteriological isolation methods. The mean number of S. Enteritidis associated with vaginal explants was significantly (P < 0.05) higher than S. Typhimurium associated with vaginal explants and both serovars associated with follicular explants. In the second experiment, the in vitro adhesion and invasion of S. Enteritidis strains in the vaginal epithelium was compared with that of several strains of S. Agona, S. Infantis, S. Hadar, S. Heidelberg, S. Montevideo and S. Typhimurium, by immunohistochemical methods. The mean number of Salmonella in the vaginal epithelium depended on their lipopolysaccharide (LPS) type, with the rank order as follows: LPS type O9 (S. Enteritidis) > LPS type O4 (S. Agona, S. Typhimurium and S. Heidelberg) > LPS type O7 (S. Montevideo and S. Infantis) and LPS type O8 (S. Hadar). This rank order of Salmonella invasiveness is in accordance with the frequency of Salmonella outbreaks involving contaminated eggs. These findings suggest that S. Enteritidis has a higher ability to colonize the vaginal epithelium than other serovars, and the Salmonella LPS type may play an essential role in tropism of the reproductive tract. PMID- 16242867 TI - Nonlinear analyses of heart rate variability in normal and growth-restricted fetuses. AB - BACKGROUND: Many studies on the physiology of the cardiovascular system reported that nonlinear chaotic dynamics may govern the generation of the heart rate signal. OBJECTIVE: To examine whether the heart rate dynamics of an intrauterine growth restricted (IUGR) fetus is different from a healthy normal fetus by nonlinear methods of time series analysis. DESIGN OF THE STUDY: One hundred nineteen fetal heart rate (FHR) recordings from healthy normal fetuses, and 69 recordings from IUGR fetuses were analyzed. Nonlinear analyses included attractor reconstruction, calculation of the largest Lyapunov exponents using the Wolf algorithm, and estimation of correlation dimension. The largest Lyapunov exponents from normal fetuses were checked by means of surrogate-data test. RESULTS: Abnormal FHR patterns of IUGR fetuses such as decreased variability and repetitive late decelerations presented a remarkably different structure in the reconstructed attractor. Surrogate data suggest that the FHR of healthy normal fetuses has unique nonlinear characteristics. The largest Lyapunov exponents were positive for all of 119 healthy normal fetuses, indicating that the FHR control system is sensitive to initial conditions. The values of IUGR fetuses were significantly lower than those of normal subjects. In normal fetuses, significant changes of correlation dimension according to gestational age were observed. In IUGR fetuses, however, such changes were not found. CONCLUSIONS: The heart rate dynamics of IUGR fetuses is less chaotic than that of normal fetuses. Decreased system complexity suggested by correlation dimension may limit the IUGR fetuses' ability to maintain cardiovascular integrity, and therefore, to adapt to the variety of internal and external cardiovascular stresses. PMID- 16242868 TI - Chemopreventive effect of punicalagin, a novel tannin component isolated from Terminalia catappa, on H-ras-transformed NIH3T3 cells. AB - Terminalia catappa and its major tannin component, punicalagin, have been characterized to possess antioxidative and anti-genotoxic activities. However, their effects on reactive oxygen species (ROS) mediated carcinogenesis are still unclear. In the present study, H-ras-transformed NIH3T3 cells were used to evaluate the chemopreventive effect of T. catappa water extract (TCE) and punicalagin. In the cell proliferation assay, TCE and punicalagin suppressed the proliferation of H-ras-transformed NIH3T3 cells with a dose-dependent manner but only partially affected non-transformed NIH3T3 cells proliferation. The differential cytotoxicity of TCE/punicalagin on the H-ras-transformed and non transformed NIH3T3 cells indicated the selectivity of TCE/punicalagin against H ras induced transformation. TCE or punicalagin treatment reduced anchorage independent growth that could be due to a cell cycle arrest at G0/G1 phase. The intracellular superoxide level, known to modulate downstream signaling of Ras protein, was decreased by punicalagin treatments. The levels of phosphorylated JNK-1 and p38 were also decreased with punicalagin treatments. Thus, the chemopreventive effect of punicalagin against H-ras induced transformation could result from inhibition of the intracellular redox status and JNK-1/p38 activation. PMID- 16242869 TI - External and internal exposure of wine growers spraying methyl parathion. AB - Organophosphates, used in agriculture, are readily absorbed through the skin. We investigated the relationship between dermal and inhalative methyl parathion exposure and the plasma levels. Twenty-three healthy wine growers sprayed the insecticide for 50 min. Fluorescent brilliant sulfoflavin was added to the spraying fluids and filter papers were fixed on the subjects. The filter papers were used to evaluate the amount of brilliant sulfoflavine on the unprotected skin fluorometrically. Inhalative exposure was measured with personal air sampler. Plasma concentrations of methyl parathion and its metabolite methyl paraoxon were determined with gas chromatography. Cholinesterase activity in serum and erythrocytes was measured before and after exposure. Some wine growers wore protective clothes, none protective gloves. Dermal exposure ranged up to 12,044 microg, inhalative to 22 microg. Maximum plasma concentration of methyl parathion was 12.1 microg/l. Methyl paraoxon was not detectable. Cholinesterase activity did not decrease. Dermal exposure correlated with the methyl parathion plasma level (Spearman's rho=0.72, p<0.001). In conclusion, dermal exposure exceeded inhalative exposure considerably. Measuring dermal deposition with the brilliant sulfoflavin technique may provide a good estimate of the internal load with methyl parathion. Preventive measures should be improved as toxic effects of repeated exposure to low doses of methyl parathion cannot be excluded. PMID- 16242870 TI - Bio-effect monitoring using a [(15)N]methacetin test as diagnostic tool to monitor remediation effects in an industrially polluted region. AB - The classical way to demonstrate the efficiency of remediation is measuring the reduction of toxic compounds in the environment. Nevertheless, more important is the risk reduction in human health. To determine changing health effects, exposure and bio-effects have to be monitored at time of and during remediation. Kindergarten children from a heavily polluted industrial (n=23) and a control area (n=12) were investigated. The region-specific outdoor and indoor exposure [27 volatile organic compounds (VOC), emphasis on tri- and tetrachloroethylene (TRI, TETRA)], the internal load [(trichloroacetic acid-TCA-as urine metabolites of TRI and TETRA and S-phenyl- and S-benzylmercapturic acid (SPMA and SBMA) as metabolites of benzene and toluene], and biological effect assessment ([(15)N]methacetin test-a non-invasive stable isotope test to determine the unspecific liver detoxification capacity of an individual) were measured twice a year during 2 years of remediation (1997/1998). It could be shown that in- and outdoor levels of TRI and TETRA decreased by 47% in the heavily polluted village, Greppin, while the levels remained much the same in the control village, Roitzsch. This trend was reflected in the decreasing elimination of TCA in the urine (41%) by the Greppin children, with no differences in the TCA elimination in Roitzsch probands. As the remediation efforts decreased the burden of exposure, the children's liver detoxification capacity improved as well. Combining different methods, such as exposure-effect (external and internal loads) and bio-effect monitoring, proved to be useful to assess remediation successes including the improvement in human health. PMID- 16242871 TI - Mercapturic acids of styrene in man: comparability of the results obtained by LC/MS/MS and by HPLC-fluorimeter, and stability of samples under different storage conditions. AB - Two analytical methods (HPLC-fluorimeter [HPLC-FLD] and tandem mass spectrometry LC/MS/MS) are available to assay phenyl-hydroxyethylmercapturic acids (PHEMAs), the mercapturic acids of styrene in humans. In the past, each method was used to check different populations of subjects, but until now no attempt has been made to compare the two methods. This study was designed to verify whether the two methods actually give comparable results. The influence of different conditions of sample storage in altering the concentration of PHEMAs was also investigated. Urine samples were collected at the beginning and at the end of the workshift from 10 workers exposed to different levels of styrene. Each sample was analysed both by LC/MS/MS after storage under different conditions (respectively, at -20 and +4 degrees C, and after repeated freezing-thawing cycles), and by HPLC-FLD (in the same conditions of storage). Strong correlations were found between the two methods both for total PHEMAs and for each of the isomers measured, including the minor (S,R)-M1. Also an alternative approach, the Bland-Altman test, confirmed the agreement between the two methods. The different storage conditions tested did not decrease the concentration of PHEMAs but, surprisingly, a clear trend to increase was shown, particularly for (R,R)-M1, (S,R)-M2 and (R,R)-M2 in samples stored at +4 degrees C for 1 week. In conclusion, the study demonstrates that the methods give comparable results. Indirectly, this confirms also the main characteristics of PHEMAs, showed in the previous experiments: low biotransformation rates of styrene into PHEMAs; large inter-individual variability; and the presence of a clear preference in the excretion of the isomers deriving from (S)-styrene oxide. PHEMAs appear stable under different storage conditions, but further studies are needed to explain the increase of levels that occurs when samples are not kept frozen. To avoid pre-analytical errors, samples collected for biomonitoring or research purposes should be frozen as soon as possible, and thawed only one time just before the analysis. PMID- 16242872 TI - The effects of Vitamin A administration on the development of vitrified-warmed mouse blastocyst. AB - The purpose of this study was to evaluate the development of vitrified-warmed mouse blastocysts following a period of Vitamin A administration. Four to six weeks old BALB/c mice were given an intraperitoneal injection of either 0.1 ml paraffin oil alone (control, Con) or paraffin oil containing 250IU of Vitamin A (experiment, Exp). Ten days later the mice were given second paraffin or paraffin Vitamin A injection and an injection of 10IU equine chorionic gonadotropin (eCG) followed 48 h later by 10IU human chorionic gonadotropin (hCG). Blastocysts were collected from both groups and randomly divided into non-vitrified (Con 1, Exp 1) and vitrified (Con 2, Exp 2) subgroups. Embryos in the vitrified group were exposed sequentially to two solutions (10% ethylene glycol, 10% DMSO in holding medium (HM: DMEMF(12)+10% FBS) and 20% ethylene glycol, 20% DMSO in HM) before plunging into liquid nitrogen. After warming at 37 degrees C, cryoprotectants were diluted serially with 0.25 and 0.15M sucrose solution in HM. The vitrified warmed and the fresh embryos of the control and the experiment groups were cultured in DMEMF(12) with 10% FBS for 72 h. Although, on the first day of culture, the rate of development to the hatched blastocyst was nearly identical between the two vitrified groups (15.8% versus 13%) but after 48 h, the rate of plated embryos was statistically higher in the vitrified Vitamin A than the vitrified control group (63.1% versus 19.6%, P<0.001). After 48 h, in the non vitrified groups, the rate of the plated embryos was also significantly higher in the Vitamin A than the control group (70.5% versus 49.3%, P<0.01). These data provided evidence that systemic administration of Vitamin A may enhance the potential development of blastocysts in culture and is capable to reduce the adverse effects of vitrification at least during the first 2 days of cultivation. PMID- 16242873 TI - Candidate gene markers for sperm quality and fertility of boar. AB - Candidate genes gonadotropin releasing hormone receptor (GNRHR), prolactin (PRL), prolactin receptor (PRLR), follicle-stimulating hormone beta (FSHB), luteinizing hormone beta (LHB), follistatin (FST), inhibin alpha (INHA), inhibin beta A (INHBA) and inhibin beta B (INHBB) were investigated for their association with sperm quality traits of sperm concentration (SCON), motility (MOT), semen volume per ejaculate (VOL), plasma droplets rate (PDR), abnormal sperm rate (ASR) and fertility traits of non return rate (NRR) and number of piglets born alive (NBA). The experimental material included 356 boars of Pietrain (PI) and Pietrain x Hampshire (PI x HA). Analysis of variance revealed significant association of GNRHR with MOT (P = 0.0161), PDR (P = 0.0048) and ASR (P = 0.0201), INHBA was found to have significant effects on PDR (P = 0.0318) and ASR (P = 0.0067), INHBB was significant (P = 0.0360) for SCON trait. FSHB, FST, INHA, PRL, PRLR and LHB had no significant effects on any trait in this experiment. PMID- 16242874 TI - ESR1 and APOE gene polymorphisms, serum lipids, and hormonal replacement therapy. AB - OBJECTIVES: The risks and benefits of hormone replacement therapy (HRT) are, at least in part, mediated by the metabolic individuality of women. Therefore, we investigated the association between polymorphisms at the estrogen receptor 1 gene (ESR1) and at the apolipoprotein E gene (APOE) with lipid and lipoprotein levels in order to verify whether these concentrations are modulated by these gene variants in women with different hormonal status. METHODS: One hundred and eighteen postmenopausal women using oral HRT with estrogen or estrogen plus progestagen (HRT+, mean age=56+/-6.7 years, 39-75 years) and 167 postmenopausal women that were not on HRT (HRT-, mean age=58+/-9.8 years, 38-85 years) participated in the study. The polymorphisms were genotyped by PCR-RFLP methods. RESULTS: No significant effect of ESR1 genotypes or haplotypes and ESR1*HRT interactions were detected on lipid levels in two-way analysis of variance. Postmenopausal women HRT nonusers carriers of the APOE*4 allele had higher T-chol and LDL-C levels than postmenopausal women HRT nonusers carriers of the APOE*3 and APOE*2 allele. T-chol and LDL-C concentrations in postmenopausal users of HRT that were APOE*4 carriers were similar to those in postmenopausal women nonusers of HRT homozygotes for APOE*3 and APOE*2 carriers. A significant APOE*4/HRT interaction was detected on T-chol and LDL-C levels by multiple regression analysis. CONCLUSION: The results from this study suggest that the HRT influence on T-chol and LDL-C levels is modulated by APOE isoforms but not by ESR1 polymorphisms. PMID- 16242876 TI - A stability-indicating HPLC method for medroxyprogesterone acetate in bulk drug and injection formulation. AB - A stability-indicating HPLC assay method has been developed and validated for medroxyprogesterone acetate (MPA) in bulk drug and injectable suspension. An isocratic RP-HPLC was achieved on a Hichrom C(18) column (150 mm x 4.6mm i.d., 5 microm) utilizing a mobile phase of methanol 0.020 M acetate buffer pH 5 (65:35, v/v) and a photodiode array detector at 245 nm. The stress testing of MPA was carried out under acidic and alkaline hydrolysis, and oxidation conditions. MPA was well resolved from its degradation products, a main related substance (megestrol acetate) and two preservatives (methyl paraben and propyl paraben) with the resolution >or=2. The proposed method was validated for selectivity, linearity, accuracy, precision and solution stability. The method was found to be suitable for the quality control of MPA in bulk drug and injections as well as the stability-indicating studies. PMID- 16242875 TI - Mixed micelles made of poly(ethylene glycol)-phosphatidylethanolamine conjugate and d-alpha-tocopheryl polyethylene glycol 1000 succinate as pharmaceutical nanocarriers for camptothecin. AB - Micelles from the mixture of poly(ethylene glycol)-phosphatidyl ethanolamine conjugate (PEG-PE) and d-alpha-tocopheryl polyetheyene glycol 1000 succinate (TPGS) were prepared loaded with the poorly soluble anticancer drug camptothecin (CPT). The solubilization of CPT by the mixed micelles was more efficient than with earlier described micelles made of PEG-PE alone. CPT-loaded mixed micelles were stable upon storage and dilution and firmly retained the incorporated drug. The cytotoxicity of the CPT-loaded mixed micelles against various cancer cells in vitro was remarkably higher than that of the free drug. PEG-PE/TPGS mixed micelles may serve as pharmaceutical nanocarriers with improved solubilization capacity for poorly soluble drugs. PMID- 16242877 TI - High sensitive analysis of rat serum bile acids by liquid chromatography/electrospray ionization tandem mass spectrometry. AB - Sensitive liquid chromatography (LC)/electrospray ionization (ESI) tandem mass spectrometry (MS) can be used to analyze the bile acid composition of rat serum. This method can analyze eight common bile acids and their glycine and taurine conjugates in 100 microl rodent serum by gradient elution on a reversed-phase column using a mixture of 20mM ammonium acetate buffer (pH 8.0), acetonitrile and methanol as a mobile phase. Selected reaction monitoring analysis under negative ion detection mode allowed the achievement of a high sensitive assay with a simple solid phase extraction using an ODS cartridge column. We used this method to investigate the effect of a one-day fast on the concentration and composition of serum bile acids in rats. The results suggested that the method described here is useful for the dynamic analysis of serum bile acids in rats. PMID- 16242878 TI - Degradation chemistry of a Vitamin D analogue (ecalcidene) investigated by HPLC MS, HPLC-NMR and chemical derivatization. AB - Ecalcidene (1-[(1alpha,3beta,5Z,7E,20S)-1,3-dihydroxy-24-oxo-9,10-secochola 5,7,10(19)-trien-24-yl]-piperidine) is a new 1-hydroxyvitamin D analogue. In this report, the thermal degradation, acid induced degradation and iodine induced degradation of ecalcidene were investigated using HPLC-MS, HPLC-NMR and chemical derivatization. In solution ecalcidene was thermally and reversibly transformed to a pre-Vitamin D type isomer 1 which subsequently produced the dehydrated pyrocalciferol and isopyrocalciferol type isomers 2 and 3 by cyclization and dehydration at elevated temperatures. Acidic conditions resulted in the formation of a novel C9-hydroxylated isomer 4 of ecalcidene, possibly via a tachysterol type intermediate, followed by the acid facilitated nucleophilic addition of water. In the presence of iodine, cis/trans isomerization of both ecalcidene and its pre-Vitamin D type isomer 1 occurred. The results may shed light on the stability and metabolism of ecalcidene, provide useful information for its potential pharmaceutical development, and enrich the knowledge of Vitamin D chemistry. PMID- 16242879 TI - APTS-labeled dextran ladder: a novel tool to characterize cell layer tightness. AB - The aim of this work was the development of an easy manageable analytic system for describing tightness of cell layers in a molecular size dependent manner, which is more precise than currently used ones. Dextrans were labeled by reductive amination with fluorescent 1-aminopyrene-3,6,8-trisulfonate (APTS). This mixture, including internal standard diazepam, was used for transport studies, which were accomplished with an established transwell blood-brain barrier model culturing an immortalized porcine brain microvascular endothelial cell line (PBMEC/C1-2). Samples were analyzed by fluorescence measurements, capillary electrophoresis and RP-LC. Following this approach, a permeability pattern could be achieved including each single fraction from APTS, APTS-glucose to APTS-dextran consisting of 31 glucose units. Permeability coefficients were calculated and ranged from 16.38+/-3.79 microm/min for APTS to 6.07+/-1.23 microm/min for the APTS-dextran with 31 glucose units (diazepam: 67.97+/-7.32 microm/min). All in all, the developed APTS-dextran ladder is an useful tool to characterize cell layer tightness--especially to describe paracellular transport ways and leakiness status of the blood-brain barrier over time--applying a wide range from smaller to larger molecules at the same time in order to refine, e.g. TEER, sucrose or Evans blue measurements. PMID- 16242881 TI - Liquid chromatography-negative ion electrospray tandem mass spectrometry method for the quantification of ezetimibe in human plasma. AB - A simple, reliable and sensitive liquid chromatography-tandem mass spectrometry method (LC-MS/MS) was developed and validated for quantification of free and total ezetimibe in human plasma. The analyte and internal standard (13C6 ezetimibe) were extracted by liquid-liquid extraction with methyl tert-butyl ether. The reversed-phase chromatographic separation was performed on a Capcell C18 column, and the plasma extract was eluted with a gradient consisting of acetonitrile and 5 mM ammonium acetate. The analyte was detected using negative ionization by multiple reaction monitoring mode. The mass transition pairs of m/z 408.5-->270.8 and m/z 414.5-->276.8 were used to detect ezetimibe and internal standard, respectively. The assay exhibited linear ranges from 0.02 to 20 ng/ml for free ezetimibe and 0.25 to 250 ng/ml for total ezetimibe in human plasma. Acceptable precision and accuracy were obtained for concentrations of the calibration standard and quality control. The validated method was successfully used to analyze human plasma samples for application in a pharmacokinetic study. PMID- 16242880 TI - GC-MS fingerprint of Pogostemon cablin in China. AB - Pogostemon cablin, originating in Malaysia and India, is cultivated in southern China including Guangdong and Hainan Province, which was called GuangHuoXiang to differentiate it from the HuoXiang of the north, the species Agastache rugosa, that it resembles. Essential oil of P. cablin mainly contributes to the pharmacological activities and the therapeutic properties of the essential oils are directly correlated with their qualitative and quantitative composition. For controlling the quality, standard fingerprint of P. cablin collected from different regions was developed by using GC-MS. Nine compounds including beta patchoulene, caryophyllene, alpha-guaiene, seychellene, beta-guaiene, delta guaiene, spathulenol, patchouli alcohol and pogostone were identified among 10 main peaks in P. cablin. Hierarchical clustering analysis based on characteristics of 10 investigated peaks in GC profiles showed that 18 samples were divided into three main clusters, patchouliol-type, pogostone-type and an interim-type, which was the one between the two chemotypes. The simulative mean chromatogram for the three types P. cablin was generated using the Computer Aided Similarity Evaluation System. The fingerprint can help to distinguish the substitute or adulterant, and further assess the differences of P. cablin grown in various areas of China. PMID- 16242882 TI - Optimization of pressurized liquid extraction for Z-ligustilide, Z butylidenephthalide and ferulic acid in Angelica sinensis. AB - Pressurized liquid extraction, one of the most promising and recent sample preparation techniques, offers the advantages of reducing solvent consumption and allowing for automated sample handling. It is being exploited in diverse areas because of its distinct advantages. However, because the extraction is performed at elevated temperatures using PLE, thermal degradation could be a concern. Z ligustilide, one of the biologically active components in Angelica sinensis, is an unstable compound, which decomposes rapidly at high temperature. In this study, we carried out a comparative study to evaluate PLE as a possible alternative to current extraction methods like Soxhlet and sonication for simultaneous extraction of Z-ligustilide, Z-butylidenephthalide and ferulic acid in A. sinensis. The operating parameters for PLE including extraction solvent, particle size, pressure, temperature, static extraction time, flush volume and numbers of extraction were optimized by using univariate approach coupled with central composite design (CCD) in order to obtain the highest extraction efficiency. Determination of Z-ligustilide, Z-butylidenephthalide and ferulic acid were carried out by means of high performance liquid chromatography with diode-array detector. The results showed that PLE was a simple, high efficient and automated method with lower solvent consumption compared to conventional extraction methods such as Soxhlet and sonication. PLE could be used for simultaneous extraction of Z-ligustilide, Z-butylidenephthalide and ferulic acid in A. sinensis. PMID- 16242883 TI - The use of LC/MS, GC/MS, and LC/NMR hyphenated techniques to identify a drug degradation product in pharmaceutical development. AB - Understanding drug degradation in the formulated product is critical in pharmaceutical development as it has significant impacts on drug efficacy, safety profile and storage conditions. As a result, identification of degradation compounds has taken an important role in the drug development process. In this study, various hyphenated analytical techniques, such as liquid chromatography mass spectrometry (LC/MS), gas chromatography mass spectrometry (GC/MS), and liquid chromatography nuclear magnetic resonance with a solid phase extraction interface (LC/SPE/NMR), have been applied to the identification of a drug degradation product which grew over time in the stability study of the drug product. The target unknown is less polar and more unsaturated than the drug substance based upon reverse phase HPLC relative retention time and UV spectra. It is not ionizable by electrospray ionization (ESI) or atmospheric pressure chemical ionization (APCI) in either a positive or a negative mode. The unknown was isolated by an HPLC fraction collector and enriched by solid phase extraction. GC/MS with chemical ionization (CI) was employed to determine the molecular weight of this compound. Its fragmentation pattern was determined by CI MS/MS using an ion trap mass spectrometer. The isolated material was also analyzed by LC/SPE/NMR, from which the structure of this compound was further characterized. The study utilizes a combination of various hyphenated analytical techniques to obtain complimentary information for structure elucidation of the unknown. The combination approach is critical for unambiguous impurity structure elucidation in drug degradation studies of pharmaceutical drug products. PMID- 16242884 TI - Development and validation of a novel LC non-derivatization method for the determination of amikacin in pharmaceuticals based on evaporative light scattering detection. AB - A novel method for the direct determination of the aminoglycoside antibiotic amikacin and its precursor component kanamycin was developed and validated, based on reversed phase LC with evaporative light scattering detector (ELSD). ELSD response to amikacin was found to be enhanced by: (a) use of ion-pairing acidic reagents of increased molecular mass, (b) increase of mobile phase volatility and (c) decrease of peak width and asymmetry (obtained by controlling the mobile phase acidity and/or ratio of organic solvent to water). Utilizing a Thermo Hypersil BetaBasic C(18) column, the selected optimized mobile phase was water methanol (60:40, v/v), containing 3.0 mll(-1) nonafluoropentanoic acid (18.2mM) (isocratic elution with flow rate of 1.0 mlmin(-1)). ELSD experimental parameters were: nitrogen pressure 3.5 bar, evaporation temperature 50 degrees C, and gain 11. Amikacin was eluted at 8.6 min and kanamycin at 10.4 min with a resolution of 1.5. Logarithmic calibration curves were obtained from 7 to 77 microgml(-1) (r>0.9995) for amikacin and 8 to 105 microgml(-1) (r>0.998) for kanamycin, with a LOD equal to 2.2 and 2.5 microgml(-1), respectively. In amikacin sulfate pharmaceutical raw materials, the simultaneous determination of sulfate (t(R)=2.3 min, LOD=1.8 microgml(-1), range 5-40 microgml(-1), %R.S.D.=1.1, r>0.9997), kanamycin and amikacin was feasible. No significant difference was found between the results of the developed LC-ELSD method and those of reference methods, while the mean recovery of kanamycin from spiked samples (0.5%, w/w) was 97.3% (%R.S.D. 148.1 and m/z 212.4--> 162.3 were used for detection of deglymidodrine and its internal standard, respectively. The assay exhibited linear ranges from 0.25 to 32 ng/ml for the analyte in human plasma. Acceptable precision and accuracy were obtained for concentrations of quality control (QC) samples. The proposed method has been successfully used to analyze human plasma samples for application in oral pharmacokinetic study. PMID- 16242886 TI - Determination of surface-bound hydroxypropylcellulose (HPC) on drug particles in colloidal dispersions using size exclusion chromatography: a comparison of ELS and RI detection. AB - Evaporative light scattering (ELS) and refractive index (RI) detection methods were evaluated for the determination of surface-bound hydroxypropylcellulose (HPC) on drug particles in colloidal dispersions. Size exclusion chromatography (SEC) was used to separate HPC from other components of the dispersions. The instrumental parameters of the ELS detector were optimized to obtain maximum peak intensity, adequate peak shape and minimal baseline noise by varying the mobile phase flow rate, nebulizer temperature, and evaporation temperature. The chromatographic method was validated using both detectors. The ELS detector response exhibited second order polynomial and linear double logarithmic correlation with concentration over a 10-300% range while the RI response was linear. The double logarithmic correlation simplified the calculation compared to using the polynomial fit, and it provided more accurate results compared to the linear fit approach. Total HPC was obtained by solubilizing all components of the dispersion and analyzing for HPC. Non-bound HPC was obtained by ultracentrifuging the dispersion and analyzing the supernatant for HPC concentration. Analysis for total- and non-bound HPC in a representative colloidal dispersion gave method precisions with R.S.D.s of 2.5 and 2.2% for ELS, and 4.5 and 2.4% for RI (n=4). HPC bound to the surface of the drug particles was determined by difference: % bound HPC=100%-% non-bound HPC. Resultant % bound HPC values ranged from 22.1 to 25.4% of available HPC. Both ELS and RI are satisfactory detection techniques for HPC quantitation and for determination of the proportion of HPC bound to drug colloid particles, and the assay results are comparable. PMID- 16242887 TI - Screening a library of potential prion therapeutics against cellular prion proteins and insights into their mode of biological activities by surface plasmon resonance. AB - The conversion of cellular prion protein (PrP(C)) to the protease resistant isoform (PrP(Sc)) is considered essential for the progression of transmissible spongiform encephalopathies (TSEs). A potential therapeutic strategy for preventing the accumulation of PrP(Sc) is to stabilize PrP(C) through the direct binding of a small molecule to make conversion less energetically favourable. Using surface plasmon resonance (SPR)-based technology we have developed a procedure, based on direct binding, for the screening of small molecules against PrP(C) immobilized on a sensor chip. In this paper we report some problems associated with the immobilization of PrP(C) onto the sensor surface for conducting drug screening and how these problems were overcome. We demonstrated that the conformational change of PrP(C) on the chip surface leads to increased exposure of the C-terminal which was observed by the increase in quinacrine binding over time, and loss of heparin binding to the N-terminal. In addition, we also report the results of the successful screening of a library of 47 compounds of known activity in cell line or cell free conversion studies for direct binding to three forms of PrP(C) (huPrP(C), t-huPrP(C) and moPrP(C)). These results show the usefulness of this technique for the identification of PrP(C) binding ligands and to gain some insight as to their potential mode of action. PMID- 16242888 TI - In vitro characterization of an in situ microdialysis sampling assay for elastase activity detection. AB - A microdialysis sampling method has been developed to detect the in vitro presence of a proteolytic enzyme, porcine elastase, external to a microdialysis probe. Elastase converts the substrate, succinyl(Ala)(3)-p-nitroanilide (suc(Ala)(3)-p-NA), to p-nitroaniline (p-NA). The substrate, suc(Ala)(3)-p-NA, was locally delivered through the microdialysis probe to external solutions containing different elastase activities (0.025-0.5 units/mL). The product, p-NA, was recovered back into the probe. Dialysates containing both suc(Ala)(3)-p-NA and p-NA were quantified using HPLC-UV. Different microdialysis suc(Ala)(3)-p-NA extraction efficiencies (EE) were observed among different elastase-containing solutions (buffer and 0.3% agar solutions). The p-NA concentrations recovered back into the microdialysis probe correlated with the elastase activity external to the microdialysis probe. The greatest fraction of p-NA recovered as compared to substrate lost occurred with the highest flow rate used (5.0 microL/min). However, the highest concentrations of p-NA recovered occurred at the lowest flow rates. This method may allow for microdialysis sampling to be used as a means to study localized enzyme activity. PMID- 16242889 TI - A study of variable hydration states in topotecan hydrochloride. AB - Topotecan hydrochloride, a pharmaceutical compound developed as a treatment for cancer, exhibits variable hydration states in a crystalline solid form chosen for manufacturing. This variability requires additional controls for successful development, and presents a characterization and detection challenge for analytical methods. In this study, overall water content was determined by Karl Fischer titration and thermogravimetric analysis (TGA) on topotecan HCl equilibrated at different relative humidity levels. These results, when combined with information obtained from dynamic water vapor sorption and differential scanning calorimetry (DSC), indicate that this form of topotecan HCl contains 3 mol of water integral to the crystalline structure and up to two additional moles of water depending on the relative humidity. Powder X-ray diffraction experiments did not detect significant differences in topotecan HCl samples equilibrated at trihydrate and pentahydrate states, and showed that the crystal lattice dimensions are not affected unless the form is dried below the trihydrate state. This behavior is typical of crystal structures with channels that can accommodate additional loosely bound water. To study the role of the loosely bound water in the crystal structure in more detail, solid-state (13)C and (15)N nuclear magnetic resonance (NMR) were used to examine the differences between the hydration states. Both the trihydrate and pentahydrate states yielded similar solid-state NMR spectra, consistent with the lack of change in the crystal lattice. However, minor but readily detectable differences in the (13)C spectra are observed with changes in water content. Interpretation of this data suggests that the loosely bound channel water is hydrogen-bonding to specific portions of the topotecan parent molecule. Topotecan HCl trihydrate was hydrated with D(2)O vapor to confirm the nature and location of the channel water using (13)C and (2)H solid-state NMR. Despite the detectable association of the channel water with hydrogen bonding sites on the topotecan molecule, (2)H quadrupolar echo experiments indicate that the channel water is highly mobile at room temperature and at -60 degrees C. PMID- 16242890 TI - Application of constant-current coulometry for estimation of plasma total antioxidant capacity and its relationship with transition metal contents. AB - Simple and express coulometric method for the evaluation of the total antioxidant capacity (TAC) of human plasma based on the reaction with electrogenerated bromine is applied. TAC of plasma from patients with different ethiology of chronic renal failure was observed. The levels of antioxidant capacity for venous and arterial plasma are authentically different (15+/-1 kCl/L versus 11.7+/-0.7 kCl/L, p<0.01). The application of Vitamin E and ximedon as an antioxidant treatment significantly increase TAC level of plasma. Free liposoluble antioxidants in plasma in alpha-tocopherol units was determined. Redox potential of plasma is measured and its correlation with lg(TAC) is obtained. Transition metal contents of Fe, Cu, Mn, Ni, and Cr in plasma of patients with chronic renal failure is significantly higher than that for a control group. Correlation analysis has shown negative linear regression between TAC value and transition metals concentration in plasma. This confirms interrelation of processes with participation of free radicals, antioxidants and transition metals as donors of electrons in chain radical processes. Moreover, it shows utility of common parameters, TAC for example, for estimation of efficiency of antioxidant defense system in living organism, in particular its antioxidant status. PMID- 16242891 TI - Determination of ceftiofur in bovine plasma by HPLC-DAD. AB - Ceftiofur sodium is a third generation broad-spectrum cephalosporin, formulated as an intramuscular injection, which is used to treat respiratory diseases in swine, ruminants and horses. The thioester bond on ceftiofur is rapidly cleaved to give desfuroylceftiofur which is further metabolized to a disulfide dimer and various desfuroylceftiofur-protein and amino acid conjugates. Methods of analysis of ceftiofur rely on cleavage by dithioerythritol to produce desfuroylceftiofur, which is further stabilized by derivatization to desfuroylceftiofur acetamide using iodoacetamide. Previous analytical methods have extracted derivatized analyte from plasma and tissue using solid-phase extraction clean-up steps followed by HPLC analysis with results reported as ceftiofur-free acid equivalents (CFAE). The simplified method presented here involves direct HPLC injection of a cleaved and derivatized sample following a protein precipitation step with calibration by external standardization and selectivity achieved based on chromatography and diode-array detection (DAD). The assay was linear over the calibration range 0.4-40 microg/ml in plasma. Intra-batch reproducibility R.S.D. was 10.3% and intra-batch sample repeatability R.S.D. was 2.1% at the 5 microg/ml level. The mean accuracy over the range of the calibration curve was -4.2% and the detection limit was 0.15 microg/ml. The assay was successfully applied to bovine plasma following intramuscular injection of ceftiofur sodium. This simplified method is suitable for pharmacokinetic applications involving ceftiofur at normal therapeutic levels. PMID- 16242892 TI - Isolation and identification of novel impurities in spironolactone. AB - Three known and five new steroidal compounds as impurities in spironolactone were isolated from the enriched mother liquor by using various chromatographic methods. Their structures were elucidated by spectrometric analysis. New compounds were characterized as 3-(3,3-dimethoxy-5 alpha,7 alpha-epidithio-17beta hydroxy-4-androstan-17 alpha-yl) propionic acid gamma-lactone (6); 3-(3-oxo-7 alpha-acetylthio-6 beta,17beta-dihydroxy-4-androsten-17 alpha-yl) propionic acid gamma-lactone (7); 7 alpha-acetylthio-17beta-20-isopropylidendioxy-21-nor-17 alpha-pregn-4-en-3-one (8); 7 alpha-acetylthio-3-oxo-pregna-4,17(20)i-dien-22-oic acid methyl ester (9) and 7 alpha-acetylthio-17-methyl-18-nor-androsta-4-en-3-one (10). PMID- 16242893 TI - Ionization of unconjugated, glycine- and taurine-conjugated bile acids by electrospray ionization mass spectrometry. AB - We investigated the effect of organic anions as spray liquid additives on the ionization efficiency of unconjugated, glycine-conjugated and taurine-conjugated bile acids under electrospray ionization conditions. Addition of organic acids influenced the ionization efficiency of whole bile acids. Use of a stronger acid reduced the peak intensity of unconjugated and glycine-conjugated bile acids, while the use of TFA, the strongest acid tested, improved the intensity of taurine conjugates. The hydroxyl group at the C-12 alpha position of cholic acid and deoxycholic acid easily underwent intra-molecular hydrogen bonding with the side chain carboxyl group, accelerating the ionization efficiency. This intra molecular hydrogen bond may also affect the formation of product ions in low energy-CID. The addition of ammonium ions to the spray liquid influenced the ionization of all bile acids, specifically enhancing the ionization efficiency of unconjugated bile acids. PMID- 16242894 TI - High-performance liquid chromatographic assay for morphine in small plasma samples: application to pharmacokinetic studies in rats. AB - In order to perform a reliable pharmacokinetic study of morphine during subchronic treatment in rats, an easy, rapid, sensitive and selective analytical method was proposed and validated. The analyte and internal standard (naloxone) were extracted from plasma samples (100 microL) by a single solid-phase extraction method prior to reverse-phase high performance liquid chromatography (HPLC) along with electrochemical detection (ED). Standard calibration graphs were linear within a range of 3.5-1,000 ng/mL (r=0.999). The intra-day coefficients of variation (CV) were in the range of 5.8-8.5% at eight concentration levels (7-1,000 ng/mL) and the inter-day coefficient of variation ranged from 7.4 to 8.8%. The intra-day assay accuracy was in the range of -5-10% and the inter-day assay accuracy ranged from 3.0 to 9.3% of relative error (RE). The limit of quantification was 3.5 ng/mL using a plasma sample of 100 microL (15.8% of CV and 12.8% of RE). Plasma samples were stable for 2 months at -20 degrees C. This method was found to be suitable for pharmacokinetic studies in rats, after subcutaneous administration of morphine (5.6 mg/kg per day) in subchronic treatment for 6 and 12 days. PMID- 16242895 TI - Solid phase extraction and liquid chromatographic determination of sildenafil and N-demethylsildenafil in rat serum with basic mobile phase. AB - HPLC method for the determination of sildenafil and its metabolite (N demethylsildenafil) in rat serum has been developed. The technique included a solid phase extraction of the serum samples on a [poly(divinylbenzene-co-N vinylpyrrolidone)] solid phase extraction sorbent. After conditioning, the cartridge was loaded with 0.5 mL of buffered serum containing internal standard. Elution was made with 1 mL of acetonitrile. After evaporation of the eluates to dryness and reconstitution with methanol, the samples were analyzed on Kromasil C18 column phase with phosphate buffer 0.05 M/acetonitrile: 54/46, pH 8. Detection was carried out using a photodiode array detector. For sildenafil and demethylsildenafil, full validation of the proposed method was provided (linearity range, calibration curves, average extraction efficiency; average intra-day and interday variabilities, limit of detection, limit of quantification, specificity). The proposed method was successfully utilised to quantify sildenafil and N-demethylsidenafil in rat serum for a pharmacokinetic study. PMID- 16242896 TI - Chest pain: communication of symptoms and history in a London emergency department. AB - OBJECTIVE: To describe patient-provider interactions for patients in an emergency department with possible acute coronary syndrome (ACS) and to generate hypotheses about how communication might contribute to sociocultural disparities in cardiac care. METHODS: Qualitative analysis of observational data. Seventy-four consecutive patients presenting between 8 a.m. and 10 p.m. over a 4-month period. RESULTS: Participants were aged 40-85 years; 58% were male; 67% were white, 18% Afro-Caribbean, and 15% South East Asian. Observations revealed significant obstacles to communication for the majority of patients. The three most prominent impediments to effective communication were: the use of leading questions to define chest pain, patient-provider conflict as a result of, and contributor to, poor communication, and frank miscommunication due to language barriers and translational difficulties. CONCLUSION: This study documents aspects of the communication process that compromise the quality of the medical history obtained in emergency department patients with suspected ACS. Accurate diagnosis relies on an interaction that weaves both the patient's and the physician's perspective into a shared understanding of events that comprise a patient's history. When diagnostic short cuts are taken to overcome educational, cultural, or language barriers in the medical interview, they may contribute to health care disparities. PRACTICE IMPLICATIONS: Physicians should take a more attentive and careful approach to patient interviewing than was observed here and should be aware of the ways in which they shape the interview through their questions and focus. Good communication skills can be effectively taught at all levels of training and practice. PMID- 16242897 TI - Patient's satisfaction and distress compared with expectations of the medical staff. AB - OBJECTIVE: Patient satisfaction is increasingly used to measure and evaluate patient treatment in hospital. The aim of this study is to assess satisfaction regarding communication, support and the degree of anxiety and depression among the patients, and to compare reports from the patients with the expectations of the staff. METHODS: In a prospective study, 176 (93%) of the patients and 52 (80%) of the nurses completed a similar questionnaire. Degree of satisfaction was measured on a five-point scale (0-4), and degree of anxiety and depression was measured with the hospital anxiety and depression scale (HAD). RESULTS: Satisfaction regarding communication with nurses (3.4+/-0.07) and physicians (3.0+/-0.08) was higher than expected by the staff (2.9+/-0.09 and 2.4+/-0.09, respectively) (p<0.01). The staff expected a higher degree of anxiety (2.3+/-0.10 versus 1.5+/-0.12) (p<0.01). The patients scored significantly higher on HAD anxiety compared with the population in general. The HAD score correlated negatively with the level of satisfaction. CONCLUSION: The patients generally showed a high degree of satisfaction with communication and support. The staff underestimated the patients' degree of satisfaction and overestimated their degree of anxiety. PRACTICE IMPLICATIONS: Knowledge of patients' degree of satisfaction is important for satisfactory communication with the staff, compliance with treatment and in order to reduce staff's concern for their care. PMID- 16242898 TI - Understanding the decisions of cancer clinical trial participants to enter research studies: factors associated with informed consent, patient satisfaction, and decisional regret. AB - OBJECTIVE: To understand the psychosocial outcomes related to decision-making processes of individuals eligible for participation in clinical trials. METHODS: Individuals eligible to participate in selected clinical trials were contacted to complete two surveys; one shortly after participants were identified, and the second 6 weeks after the first survey was completed (N=50). Measures included subjective informed consent; satisfaction with decision-making; decisional regret; and timing of consent (early versus late signers). ANOVA and correlation coefficients were used to test the relationships between variables. RESULTS: Early signers reported themselves to be less informed about the details of their particular clinical trials than later signers (M=81.9 versus 91.2; F=5.5; p=.02). There was a non-significant trend for early signers to be less satisfied with their decisions than late signers. Satisfaction with decision-making and subjective informed consent were both strongly associated with later decisional regret (r=-.32 and -.30, respectively). However, there was no relationship between timing of consent and decisional regret. CONCLUSION: Participants who enroll in clinical trials quickly may not believe they fully understand the implications of trial participation. In general, participants who do not believe they fully understand the implications of trial participation, or who are less satisfied with their decision to enroll in the trial may ultimately feel regret about their decision to participate. PRACTICE IMPLICATIONS: More effort is needed to ensure that clinical trial participants fully understand the risks and benefits of participation and are satisfied with their decision to enroll in a trial prior to signing consent forms. PMID- 16242899 TI - Informed decision making in the context of prenatal screening. AB - OBJECTIVE: This study aimed to construct a measure of informed decision making that includes knowledge, deliberation, and value-consistency, and to assess the level of informed decision making about prenatal screening, and differences between test acceptors and test decliners. METHODS: Women attending one of 44 midwifery and gynaecology practices were asked to fill out postal questionnaires before and after the prenatal screening offer. The principal outcome was the level of informed decision making. For this purpose, knowledge about prenatal screening, deliberation about the pros and cons of the alternatives, test uptake, and attitude towards having a prenatal screening test were measured. RESULTS: Eighty-four percent of the participants were sufficiently knowledgeable about prenatal screening, 75% of the decisions were deliberate, and 82% were value consistent. Fifty-one percent of the participants made an informed decision. Test acceptors made less informed decisions as compared to test decliners. This difference was mainly caused by the lower rate of deliberation in this group. CONCLUSION: It appears from this study that prenatal screening decisions are often not informed decisions. This is inconsistent with the main objective of offering screening, which is to enable people to make informed decisions. PRACTICE IMPLICATIONS: Decision makers should be encouraged during the counselling to deliberate about the various alternatives. PMID- 16242900 TI - Transfer of communication skills training from workshop to workplace: the impact of clinical supervision. AB - OBJECTIVE: Recent studies have recognised that the communication skills learned in the training environment are not always transferred back into the clinical setting. This paper reports a study which investigated the potential of clinical supervision in enhancing the transfer process. METHODS: A randomised controlled trial was conducted involving 61 clinical nurse specialists. All attended a 3-day communication skills training workshop. Twenty-nine were then randomised to 4 weeks of clinical supervision, aimed at facilitating transfer of newly acquired skills into practice. Assessments, using real and simulated patients, were carried out before the course, immediately after the supervision period and 3 months later. Interviews were rated objectively using the Medical Interview Aural Rating Scale (MIARS) to assess nurses' ability to use key skills, respond to patient cues and identify patient concerns. RESULTS: Assessments with simulated patients showed that the training programme was extremely effective in changing competence in all three key areas. However, only those who experienced supervision showed any evidence of transfer. Improvements were found in the supervised groups' use of open questions, negotiation and psychological exploration. Whilst neither group facilitated more disclosure of cues or concerns, those in the experimental group responded more effectively to the cues disclosed, reduced their distancing behaviour and increasing their exploration of cues. CONCLUSIONS: The study has shown that whilst training enhances skills, without intervention, it may have little effect on clinical practice. The potential role of clinical supervision as one way of enhancing the clinical effectiveness of communication skills training programmes has been demonstrated. PRACTISE IMPLICATIONS: This study raises questions about the effectiveness of training programmes which do not incorporate a transfer element, and provides evidence to support the need for clinical supervision for clinical nurse specialist. PMID- 16242901 TI - Communication strategies and interpersonal skills of instructors of esophageal speech: an observation study. AB - OBJECTIVE: In this article, the way that the Instructor of Esophageal Speech (IES) works is considered from an interactive-analytic perspective. METHODS: The observation project data were gathered during 6 months in 2004 at the training sessions of the Ginreikai-Asian Federation of Laryngectomees' Association using the Six Category Intervention Analysis framework (6CIA) by a trained communication specialist as an active-observer. RESULTS: Mean scores of each of the six categories and a series of percentage distributions were extracted from the usage-frequency data and interpreted together with the results of Chi-square analysis of usage-frequency units. We found the IES used more frequently authoritative categories and used less facilitative categories. Our results of the data analysis show that the 30 IES used more authoritative interventions and used more frequently the prescription and information giving than the confrontation or cathartic interventions. CONCLUSION: The results of the present study show that the 6CIA framework has its potential and value as an analytic tool to explore the IES' actual behavior in a specific therapy related context. PRACTICE IMPLICATIONS: We argue for the utility of the 6CIA as an analytic framework to investigate the interpersonal behavior of the IES in the Japanese cultural setting. PMID- 16242903 TI - The effect of salutogenic treatment principles on coping with mental health problems A randomised controlled trial. AB - OBJECTIVE: Although the theory of salutogenesis provides generic understanding of how coping may be created, this theoretical perspective has not been explored sufficiently within research among people suffering from mental health problems. The aim of this study is to investigate the effect of talk-therapy groups based on salutogenic treatment principles on coping with mental health problems. METHOD: In an experimental design, the participants (residents in the community) were randomly allocated to a coping-enhancing experimental group (n=59) and a control group (n=47) receiving standard care. Coping was measured using the sense of coherence (SOC) questionnaire. RESULTS: Coping improved significantly in the experiment group (+6 points) compared with the control group (-2 points). The manageability component contributed most to this improvement. CONCLUSION: Talk therapy groups based on salutogenic treatment principles improve coping among people with mental health problems. PRACTICE IMPLICATIONS: Talk-therapy groups based on salutogenic treatment principles may be helpful in increasing coping in the recovery process among people with mental health problems and seem to be applicable to people with various mental health problems. PMID- 16242905 TI - Performance on the Hayling and Brixton tests in older adults: norms and correlates. AB - The individualized nature of the aging process underlines the need to have neuropsychological tests that are sensitive enough to distinguish normal changes associated with aging from those that are pathological. However, these measures are only useful if adequate normative data are available. Normative data are presented for two new executive functioning tasks, the Hayling and Brixton tests, which were administered as part of a neuropsychological battery to 457 typically aging older adults (53-90 years). Advancing age was associated with poorer performance on both the Hayling and Brixton tests. Results showed that fluid intelligence accounts for some but not all of the age-related variance on these tasks. PMID- 16242904 TI - Expressing medicine side effects: assessing the effectiveness of absolute risk, relative risk, and number needed to harm, and the provision of baseline risk information. AB - OBJECTIVE: To assess the effectiveness of absolute risk, relative risk, and number needed to harm formats for medicine side effects, with and without the provision of baseline risk information. METHODS: A two factor, risk increase format (relative, absolute and NNH)xbaseline (present/absent) between participants design was used. A sample of 268 women was given a scenario about increase in side effect risk with third generation oral contraceptives, and were required to answer written questions to assess their understanding, satisfaction, and likelihood of continuing to take the drug. RESULTS: Provision of baseline information significantly improved risk estimates and increased satisfaction, although the estimates were still considerably higher than the actual risk. No differences between presentation formats were observed when baseline information was presented. Without baseline information, absolute risk led to the most accurate performance. CONCLUSION: The findings support the importance of informing people about baseline level of risk when describing risk increases. In contrast, they offer no support for using number needed to harm. PRACTICE IMPLICATIONS: Health professionals should provide baseline risk information when presenting information about risk increases or decreases. More research is needed before numbers needed to harm (or treat) should be given to members of the general populations. PMID- 16242906 TI - Quality of life in multiple sclerosis: does information-processing speed have an independent effect? AB - Information-processing speed (IPS) has been identified as an area of primary deficit in multiple sclerosis regardless of disease course. This study examines the extent to which information-processing speed contributes to quality of life (measured by the SF-36) in individuals with Multiple Sclerosis (MS), independent of level of neurological disability (measured by the Expanded Disability Status Scale (EDSS)). Fifty-two individuals with MS (29 relapsing-remitting and 23 primary-progressive) completed the SF-36 and neuropsychological measures related to speed of processing and were assessed using the EDSS. The EDSS was significantly related to all SF-36 scales except those measuring Mental Health and Role Limits--Emotional. While the EDSS contributed significantly to prediction of SF-36 component scores, addition of measures of IPS to the regression equation did not significantly improve prediction. It was noted, however, the effect size associated with addition of IPS scores were large, indicating that quality of life is indeed related to measures of IPS. PMID- 16242907 TI - The phylogenetic analysis of tetraspanins projects the evolution of cell-cell interactions from unicellular to multicellular organisms. AB - In animals, the tetraspanins are a large superfamily of membrane proteins that play important roles in organizing various cell-cell and matrix-cell interactions and signal pathways based on such interactions. However, their origin and evolution largely remain elusive and most of the family's members are functionally unknown or less known due to difficulties of study, such as functional redundancy. In this study, we rebuilt the family's phylogeny with sequences retrieved from online databases and our cDNA library of amphioxus. We reveal that, in addition to in metazoans, various tetraspanins are extensively expressed in protozoan amoebae, fungi, and plants. We also discuss the structural evolution of tetraspanin's major extracellular domain and the relation between tetraspanin's duplication and functional redundancy. Finally, we elucidate the coevolution of tetraspanins and eukaryotes and suggest that tetraspanins play important roles in the unicell-to-multicell transition. In short, the study of tetraspanin in a phylogenetic context helps us understand the evolution of intercellular interactions. PMID- 16242908 TI - Evolutionarily conserved coupling of transcription and alternative splicing in the EPB41 (protein 4.1R) and EPB41L3 (protein 4.1B) genes. AB - Recent studies have shown that transcription and alternative splicing can be mechanistically coupled. In the EPB41 (protein 4.1R) and EPB41L3 (protein 4.1B) genes, we showed previously that promoter/alternative first exon choice is coupled to downstream splicing events in exon 2. Here we demonstrate that this coupling is conserved among several vertebrate classes from fish to mammals. The EPB41 and EPB41L3 genes from fish, bird, amphibian, and mammal genomes exhibit shared features including alternative first exons and differential splice acceptors in exon 2. In all cases, the 5'-most exon (exon 1A) splices exclusively to a weaker internal acceptor site in exon 2, skipping a fragment designated as exon 2'. Conversely, alternative first exons 1B and 1C always splice to the stronger first acceptor site, retaining exon 2'. These correlations are independent of cell type or species of origin. Since exon 2' contains a translation initiation site, splice variants generate protein isoforms with distinct N-termini. We propose that these genes represent a physiologically relevant model system for mechanistic analysis of transcription-coupled alternative splicing. PMID- 16242909 TI - Metabotropic glutamate receptors mediate lipopolysaccharide-induced fever and sickness behavior. AB - Several mechanisms have been proposed for neuroimmune communication supporting the sickness syndrome (fever, anorexia, inactivity, and cachexia) following infection. We examined the role of glutamate as a neurochemical intermediary of sickness behavior induced by intraperitoneal lipopolysaccharide (LPS). Mice implanted with biotelemetry devices capable of detecting body temperature (Tb) were administered LPS (50 or 500 microg/kg i.p., serotype 0111:B4) with or without i.p. pretreatment with vehicle or broad-spectrum antagonists selective for N-methyl-d-aspartate (NMDA), alpha-amino-3-hydroxy-5-methyl-4 isoxazoleproprionic (AMPA)/kainite, or metabotropic glutamate (mGlu) receptors. While NMDA and AMPA/kainate receptor antagonism failed to attenuate LPS-induced sickness behavior, antagonism of metabotropic receptors with l(+)-AP3 reduced the febrile (0-11h: control: 37.32+/-0.16 degrees C, l(+)-AP3: 36.66+/-0.27), anorexic (control: -87+/-5%, l(+)-AP3: 48+/-12% scotophase food intake), and cachexic (control: -8.9+/-0.4%, l(+)-AP3: -6.1+/-1.3% body weight) effects of 500 microg/kg LPS, and produced a biphasic Tb effect in response to 50 microg/kg LPS (1h: -0.90+/-0.26; 6h: 1.78+/-0.35 degrees C relative to baseline). At this dose the Tb of l(+)-AP3-treated mice was 1.18 degrees C lower than controls 2h post injection, and 0.68 degrees C greater that controls 8h post-injection. These results suggest a role for mGlu receptors in mediating fever, anorexia, and cachexia possibly via activation of extra-vagal pathways, since the attenuating effect of l(+)-AP3 increased with increasing dosages of LPS. Given the critical role ascribed to mGlu receptors in neurotransmitter release and astrocytic processes, it is possible that these observations reflect an l(+)-AP3-induced attenuation of these systems. PMID- 16242910 TI - Belief or Need? Accounting for individual variations in the neurochemistry of the placebo effect. AB - The activation of pain-suppressive, endogenous opioid neurotransmission after administration of a placebo with expectation of analgesia has been directly demonstrated in humans using molecular imaging techniques in recent work. Regional effects were described in the dorsolateral prefrontal cortex, pregenual anterior cingulate, anterior insula, and nucleus accumbens. However, it was also observed that the magnitude of these responses was subject to substantial individual and regional variation. The present study was undertaken to examine the contribution of various factors to the observed variability in the neurochemical responses to placebo administration. Multiple regression analyses were conducted on data from 19 healthy males to study to what degree expectations of analgesia and various elements of the experience of pain itself, in the absence of placebo, were associated with the individual and brain regional variability in endogenous opioid neurochemical responses to placebo. A model that included affective qualities of pain, the volume of algesic stimulus required to maintain pain over the experimental period within a moderate range, and the internal affective state of the volunteers contributed to 40-68% of the variance in the regional neurochemical responses to placebo. These initial data suggests that in the case of endogenous opioid mediated placebo analgesic responses, the individual experience of pain, in particular its affective elements, the internal affective state of the individuals during pain and a measure of sustained pain sensitivity are important factors contributing to the formation of a placebo effect. Further examination of individual variations in placebo responding will need to take into account the underlying process for which relief is required. PMID- 16242911 TI - The effects of graduated exposure, modeling, and contingent social attention on tolerance to skin care products with two children with autism. AB - Children with autism may display unusual or fearful responses to common stimuli, such as skin care products. Parents of children with autism have often reported that their children will not allow the application of these types of substances to their skin and if the parent persists, the children become extremely upset and anxious. Such responding can interfere with adaptive functioning. The purpose of this study was to evaluate the effects of a treatment package involving graduated exposure to steps in an avoidance hierarchy, modeling, and social attention on the responding of two children with autism who displayed fearful responses to skin care products. Both avoidance and acceptance responses to skin care products were measured. Both changing criteria and multiple baseline experimental designs were employed to assess the effects of the intervention package. The results suggest that the package was successful in teaching tolerance of skin products for both children. PMID- 16242912 TI - Autoimmune hepatitis type 1 and primary biliary cirrhosis have distinct bone marrow cytokine production. AB - We have recently reported differences in the hematopoiesis between autoimmune hepatitis type 1 (AIH-1) and primary biliary cirrhosis (PBC). In view of the notion that cytokines are regulators of hematopoiesis, we investigated in our tertiary center the cytokine production in the bone marrow (BM) of the same consecutive cohort of patients (13 AIH-1, 13 PBC, 10 healthy and 7 patients with cirrhosis due to chronic hepatitis B). Interferon-gamma (IFN-gamma), interleukin 4 (IL-4), interleukin-10 (IL-10), tumor necrosis factor-alpha (TNF-alpha) and transforming growth factor-beta (TGF-beta) were determined in the supernatants of long-term BM cultures by ELISAs. IL-4, TNF-alpha and TGF-beta were found significantly increased in the BM of PBC patients compared to AIH-1 and both control groups. AIH-1 patients had significantly higher BM IL-10 compared to PBC patients and higher IL-10, IL-4 and TNF-alpha compared to controls. BM IFN-gamma was significantly higher in PBC and AIH-1 patients compared to controls. In AIH-1 patients, IL-10 was positively correlated with CD34+, CD34+/CD38- and CD34+/CD38+ cell proportions. In conclusion, the BM cytokine microenvironment of PBC and AIH 1 patients differs significantly compared to that of healthy individuals and cirrhotic patients of non-autoimmune etiology. Differences were also found between patients with PBC and AH-1. The implication of BM in the pathogenesis of autoimmune liver diseases is possible and needs further investigation. PMID- 16242913 TI - Warm autoimmune hemolytic anemia is an IgM-IgG immune complex disease. AB - Warm autoimmune hemolytic anemia (WAIHA) is characterized by polyclonal IgG autoantibodies binding to red blood cells (RBC). The characterization of the autoantigen in WAIHA has not yet led to definitive results, and the etiology of RBC autoantibodies remains unclear. An altered control of self-reactive IgG by autologous IgM has been proposed as the underlying mechanism of disease in WAIHA, suggesting that IgM-IgG immune complexes contribute to the pathophysiology of the disease. In the present study, we purified and characterized IgM from plasma of WAIHA patients and from healthy controls using FPLC-based protocols and optical biosensor technology, and investigated IgG present within the IgM fractions. We provide evidence that IgM-IgG immune complexes in plasma and associated with the RBC membrane are the characteristic feature of WAIHA, independent of the etiology of the disease. IgM-IgG immune complexes of WAIHA patients differ from IgM-IgG immune complexes of healthy individuals with regard to quantity and to structural composition. The data suggest that self-immunoglobulin is the original autoantigen underlying WAIHA. The molecular characterization of IgM-IgG immune complexes may define new targets for therapeutic intervention in WAIHA. PMID- 16242914 TI - BAFF is elevated in serum of patients with Wegener's granulomatosis. AB - BAFF (B-cell activating factor of the TNF family) plays a crucial role in B-cell survival. Elevated BAFF serum levels have been linked to several autoimmune diseases in humans, and therapies targeting BAFF were successful in animal models of rheumatoid arthritis and systemic lupus erythematosus. Wagener's granulomatosis (WG), a chronic systemic vasculitis, is characterized by circulating autoantibodies (cANCA) targeting neutrophils, which can produce BAFF. To investigate whether BAFF is involved in WG pathology, BAFF serum levels were measured by ELISA in 46 WG patients and 62 healthy donors. We report the novel finding that in WG patients serum levels of BAFF were significantly increased (median 3.95 ng/ml, p=0.009) compared to healthy controls (median 2.38 ng/ml). The difference was even more pronounced when comparing controls with untreated WG patients (median 4.61 ng/ml, p=0.001). Treatment of WG patients with glucocorticoids was associated with lower BAFF levels. The serum BAFF level in treated WG patients was about the same as in the control group. We propose that BAFF might be a pathogenic factor in WG and that targeting BAFF may represent a new therapeutic strategy in a subset of chronically relapsing WG patients with elevated BAFF levels. PMID- 16242915 TI - PKCeta is localized in the Golgi, ER and nuclear envelope and translocates to the nuclear envelope upon PMA activation and serum-starvation: C1b domain and the pseudosubstrate containing fragment target PKCeta to the Golgi and the nuclear envelope. AB - Protein kinase C (PKC) represents a family of serin/threonine kinases, playing a central role in the regulation of cell growth, differentiation and transformation. These enzymes differ in their primary structure, biochemical properties, tissue distribution and subcellular localization. The specific cellular functions of PKC isoforms are largely controlled by their localization. PKCeta, a member of the novel subfamily, is expressed predominantly in epithelial tissues. However, not much is known with respect to its mechanism of activation and regulation. Our recent studies suggest its role in cell cycle control. Here we show that PKCeta is localized at the Golgi apparatus, ER and the nuclear envelope. Furthermore, using GFP-fusion proteins of the different functional domains of PKCeta we deciphered the specific structural domains of the protein responsible for its apparent localization. We show that the cysteine-rich repeat C1b is responsible for its Golgi localization, while for its presence at the ER/nuclear envelope the pseudosubstrate containing fragment coupled to the C1 domain is required. In response to short-term activation by PMA we show translocation of PKCeta to the plasma membrane and the nuclear envelope. We demonstrate that the C1b is sufficient for its translocation to the plasma membrane. Interestingly, accumulation of PKCeta at the nuclear envelope also occurred in response to serum-starvation. It should be noted that interaction of PKCeta with the cyclin E/Cdk2 complex at the perinuclear region was recently reported by us in response to serum-starvation. Thus, our studies demonstrate translocation of PKCeta to the nuclear envelope, and suggest that the spatial regulation of PKCeta could be important for its cellular functions including effects on cell cycle control and involvement in tumor promotion. PMID- 16242916 TI - ERK is an anti-inflammatory signal that suppresses expression of NF-kappaB dependent inflammatory genes by inhibiting IKK activity in endothelial cells. AB - Unveiling of endothelial nuclear factor-kappaB (NF-kappaB) activation is pivotal for understanding the inflammatory reaction and the pathogenesis of inflammatory vascular diseases. We here report the novel function of extracellular signal related kinase (ERK) in controlling endothelial NF-kappaB activation and inflammatory responses. In human endothelial cells, vascular endothelial growth factor (VEGF) induced NF-kappaB-dependent transcription of cell adhesion molecules (CAMs) and monocyte adhesion. These effects were prominently enhanced by either pretreatment with the MEK inhibitors, PD98059 and U0126 or overexpression of a dominant negative form of MEK, but blocked by a wild type ERK. Consistently, inhibition of ERK significantly increased IkappaB kinase (IKK) activity, IkappaBalpha phosphorylation, and nuclear translocation of NF-kappaB induced by VEGF, whereas overexpression of ERK resulted in the loss of these responses to VEGF. Using two PKC inhibitors has demonstrated that VEGF concomitantly stimulates IKK and its negative regulatory signal ERK through PKC that lies downstream of KDR/Flk-1. Strikingly, elevation of ERK in endothelial cells markedly inhibited CAM expression and NF-kappaB activation as well as monocyte adhesion induced by IL-1beta and TNF-alpha. The data collectively suggest that ERK serves as an anti-inflammatory signal that suppresses expression of NF-kappaB-dependent inflammatory genes by inhibiting IKK activity in endothelial cells. Measuring the existence of ERK activity in vascular endothelial cells may be useful for predicting the feasibility and potency of inflammatory reactions in the vasculature. PMID- 16242917 TI - Season of birth is associated with anthropometric and neurocognitive outcomes during infancy and childhood in a general population birth cohort. AB - The 'season of birth' effect is one of the most consistently replicated associations in schizophrenia epidemiology. In contrast, the association between season of birth and development in the general population is relatively poorly understood. The aim of this study was to explore the impact of season of birth on various anthropometric and neurocognitive variables from birth to age seven in a large, community-based birth cohort. A sample of white singleton infants born after 37 weeks gestation (n = 22,123) was drawn from the US Collaborative Perinatal Project. Anthropometric variables (weight, head circumference, length/height) and various measures of neurocognitive development, were assessed at birth, 8 months, 4 and 7 years of age. Compared to summer/autumn born infants, winter/spring born infants were significantly longer at birth, and at age seven were significantly heavier, taller and had larger head circumference. Winter/spring born infants were achieving significantly higher scores on the Bayley Motor Score at 8 months, the Graham-Ernhart Block Test at age 4, the Wechsler Intelligence Performance and Full Scale scores at age 7, but had significantly lower scores on the Bender-Gestalt Test at age 7 years. Winter/spring birth, while associated with an increased risk of schizophrenia, is generally associated with superior outcomes with respect to physical and cognitive development. PMID- 16242918 TI - Premorbid childhood ocular alignment abnormalities and adult schizophrenia spectrum disorder. AB - This study examined the relation between childhood ocular alignment deficits and adult psychiatric outcomes among children at high-risk for schizophrenia and controls. A sample of 265 Danish children was administered a standardized eye exam assessing strabismus and related ocular alignment deficits. All children whose mothers or fathers had a psychiatric diagnosis of schizophrenia comprised the first group (N=90). Children who had at least one parent with a diagnosis other than schizophrenia comprised the first matched control group (N=93). The second control group consisted of children with no parental diagnoses (N=82). In 1992, adult psychiatric outcome data were obtained for 242 of the original subjects. It was found that children who later developed a schizophrenia-spectrum disorder had significantly higher eye exam scale and strabismus scale scores compared to children who developed other non-psychotic psychopathology and children who did not develop a mental illness. The mean rank for children in the high-risk group (offspring of parents with schizophrenia) on the eye scale and the strabismus scale was greater than the mean rank for children in the matched control groups (both offspring of parents with other non-psychotic disorder and no mental illness), although the results failed to reach statistical significance. Results from this study suggest a premorbid relation between ocular deficits and schizophrenia-spectrum disorders in childhood prior to onset of psychopathology in adulthood. Strabismus may serve as a premorbid marker for spectrum disorders and may have implications for the understanding of early aberrant neurological development related to later schizophrenia-spectrum disorders. PMID- 16242919 TI - Dopaminergic drugs may counteract behavioral and biochemical changes induced by models of brain injury. AB - The dopaminergic drugs, bromocriptine, cabergoline, dihydroergocryptine, pergolide and ropinirole were injected subcutaneously (s.c.) at the dose of 0.1, 0.5 and 1 mg/kg/day for 7 days into male rats of the Sprague-Dawley strain. The drug pre-treatment reverted amnesia induced in rats by hypobaric hypopxia and tested in active and passive avoidance tasks. A restoration of memory retention, as assessed in a step-through passive avoidance task, was found in animals with a 2-month brain occlusive ischemia and exposed to dopaminergic drugs for 7 days. For behavioral effects in both active and passive avoidance tests in both experimental models, the rank of relative potency was ropirinole>bromocriptine=cabergoline>pergolide>dihydroergocryptine. Spontaneous ambulation of animals with brain occlusive ischemia was increased by the higher doses of drugs. All dopaminergic drugs reduced kainate mortality rate. The rank of relative potency for this effect was ropirinole=bromocriptine=cabergoline>pergolide=dihydroergocryptine. However, no change was found in other seizure parameters (latency to first convulsion and total number of convulsions) after drug treatment. A biochemical analysis of glutathione redox index (glutathione reduced/glutathione oxidized ratio) in discrete brain areas revealed that exposure to dopaminergic drugs increased this parameter in frontal cortex, striatum and hippocampus of animals subject to hypobaric hypoxia and brain occlusive ischemia. For this effect, the relative potency rank was ropirinole>bromocriptine=cabergoline>>pergolide=dihydroergocryptine. These behavioral and biochemical findings suggest that dopaminergic drugs may counteract either behavioral or biochemical changes induced by experimental models of brain injury. This activity was found after protective activity (as found in animals pre-treated with these drugs and exposed to hypobaric hypoxia) or reversal of brain injury (as found in animals treated after 2-month occlusive brain ischemia). Their neuroprotective activity probably involves the reduction/oxidation balance of the glutathione system in the brain. PMID- 16242920 TI - Distinct gene expression profiles in adult rat brains after acute MK-801 and cocaine treatments. AB - Uncompetitive N-methyl-D-aspartate (NMDA) receptor antagonists have been suggested to attenuate the self-administration and rewarding effects of psychostimulants. Microarrays containing 14,500 rat cDNAs were hybridized to identify alterations in gene expression levels in rat brain regions elicited by the uncompetitive NMDA receptor antagonist MK-801 (dizocilpine, 1 mg/kg), the dopamine agonist cocaine (20 mg/kg), or combined treatment (MK-801 15 min prior to cocaine) 4 h after injections. Total genes up regulated (Z-ratio >2) in parietal cortex and nucleus accumbens were 111 and 158, respectively. Total genes down regulated (Z-ratio <2) in the same tissues were 360 and 166, respectively. These genes fell into multiple molecular function gene ontology (GO) categories, but were highly represented in catalytic activities (44% of all genes), signal transduction (14%), protein (20%), nucleotide (18%), and nucleic acid (15%) binding. In nucleus accumbens, genes up regulated by MK-801 (87 genes) did not overlap those up regulated by cocaine (46 genes). Genes down regulated by MK-801 (33 genes) consisted of 2 overlapping genes with those down regulated by cocaine (89 genes). In parietal cortex, low numbers of overlapping regulated genes were also observed. Combined treatments also indicated low numbers (0-10) of genes commonly regulated when compared with single treatments alone. In situ hybridisation studies indicated significant increases in b-ZIP transcription factors (CREM, ICER, CBP, and c-fos) elicited by MK-801 and decreases in c-fos elicited by cocaine. The results indicate independent gene expression signatures following acute MK-801 and cocaine administration that appears to be largely non overlapping and context dependent. PMID- 16242921 TI - Mitochondrial criticality: a new concept at the turning point of life or death. AB - A variety of stressors can cause the collapse of mitochondrial membrane potential (DeltaPsi(m)), but the events leading up to this catastrophic cellular event are not well understood at the mechanistic level. Based on our recent studies of oscillations in mitochondrial energetics, we have coined the term "mitochondrial criticality" to describe the state in which the mitochondrial network of cardiomyocytes becomes very sensitive to small perturbations in reactive oxygen species (ROS), resulting in the scaling of local mitochondrial uncoupling and DeltaPsi(m) loss to the whole cell, and the myocardial syncytium. At the point of criticality, the dynamics of the mitochondrial network bifurcate to oscillatory behavior. These energetic changes are translated into effects on the electrical excitability of the cell, inducing dramatic changes in the morphology and the threshold for activating an action potential. Emerging evidence suggests that this mechanism, by creating spatial and temporal heterogeneity of excitability in the heart during ischemia and reperfusion, underlies the genesis of potentially lethal cardiac arrhythmias. PMID- 16242922 TI - Double dissociation in neural correlates of visual working memory: a PET study. AB - Using positron emission tomography (PET), we investigated the organisation of spatial versus object-based visual working memory in 11 normal human subjects. The paradigm involved a conditional colour-response association task embedded within two visual working memory tasks. The subject had to remember a position (spatial) or shape (object-based) and then use this to recover the colour of the matching element for the conditional association. Activation of the nucleus accumbens and the anterior cingulate cortex was observed during the conditional associative task, indicating a possible role of these limbic structures in associative memory. When the 2 memory tasks were contrasted, we observed activation of 2 distinct cortical networks: (1) The spatial task activated a dorsal stream network distributed in the right hemisphere in the parieto occipital cortex and the dorsal prefrontal cortex, and (2) The non spatial task activated a ventral stream network distributed in the left hemisphere in the temporo- occipital cortex, the ventral prefrontal cortex and the striatum. These results support the existence of a domain-specific dissociation with dorsal and ventral cortical systems involved respectively in spatial and non spatial working memory functions. PMID- 16242923 TI - Neuroimaging studies of practice-related change: fMRI and meta-analytic evidence of a domain-general control network for learning. AB - Functional magnetic resonance imaging and a meta-analysis of prior neuroimaging studies were used to characterize cortical changes resulting from extensive practice and to evaluate a dual-processing account of the neural mechanisms underlying human learning. Three core predictions of the dual processing theory are evaluated: 1) that practice elicits generalized reductions in regional activity by reducing the load on the cognitive control mechanisms that scaffold early learning; 2) that these control mechanisms are domain-general; and 3) that no separate processing pathway emerges as skill develops. To evaluate these predictions, a meta-analysis of prior neuroimaging studies and a within-subjects fMRI experiment contrasting unpracticed to practiced performance in a paired associate task were conducted. The principal effect of practice was found to be a reduction in the extent and magnitude of activity in a cortical network spanning bilateral dorsal prefrontal, left ventral prefrontal, medial frontal (anterior cingulate), left insular, bilateral parietal, and occipito-temporal (fusiform) areas. These activity reductions are shown to occur in common regions across prior neuroimaging studies and for both verbal and nonverbal paired-associate learning in the present fMRI experiment. The implicated network of brain regions is interpreted as a domain-general system engaged specifically to support novice, but not practiced, performance. PMID- 16242924 TI - Action selectivity in parietal and temporal cortex. AB - The sensory-action theory proposes that the neural substrates underlying action representations are related to a visuomotor action system encompassing the left ventral premotor cortex, the anterior intraparietal (AIP) and left posterior middle temporal gyrus (LPMT). Using fMRI, we demonstrate that semantic decisions on action, relative to non-action words, increased activation in the left AIP and LPMT irrespective of whether the words were presented in a written or spoken form. Left AIP and LPMT might thus play the role of amodal semantic regions that can be activated via auditory as well as visual input. Left AIP and LPMT did not distinguish between different types of actions such as hand actions and whole body movements, although a right STS region responded selectively to whole body movements. PMID- 16242925 TI - Release property of temperature-sensitive alginate beads containing poly(N isopropylacrylamide). AB - The graft copolymer (APN) of alginate and poly(N-isopropylacrylamide) (PNIPAM) were synthesized and APN beads were prepared by dropping the aqueous solution of the copolymer into an aqueous solution of Ca(2+) solution. Alginate chains were employed to play a role in forming beads by electrostatic interactions with a multivalent ion, Ca(2+). Grafted PNIPAM segments were adopted to act as a valve for the pores of the beads, since they exhibit the properties of thermal contraction and expansion. The percent of release of blue dextran from APN beads was higher at 40 degrees C than at 25 degrees C. The difference in the release between two temperatures became more distinguishable when the content of PNIPAM in APN beads is higher. Below lower critical solution temperature (LCST), the expanded PNIPAM would close the pores of the beads, resulting in a lower release rate. Above LCST, the thermally contracted polymer would open the pores, resulting in a higher release rate. The percent of release from APN beads were investigated when the temperature of the release medium is altered. The release rate was relatively low at 25 degrees C. The temperature, however, changed up to 40 degrees C, a marked increase in the release rate was observed. These trends were found to be reproducible when the temperature was repeatedly altered between 25 and 40 degrees C. As a result, a stepwise response to the temperature alteration was obtained. PMID- 16242926 TI - Long-term oxygen therapy in chronic respiratory failure: a Multicenter Italian Study on Oxygen Therapy Adherence (MISOTA). AB - BACKGROUND: The adherence to the prescribed oxygen therapy is difficult to obtain for patients on long-term oxygen therapy (LTOT). There is little information on the modalities of oxygen utilisation for patients on LTOT who are using liquid oxygen in real life. STUDY OBJECTIVE: Evaluation of the behaviour and the knowledge regarding LTOT in a large group of patients mainly using liquid oxygen. DESIGN AND SETTING: Questionnaire administered to consecutive outpatients on domiciliary LTOT for at least 6 months referring to one of 20 clinics throughout Italy. Blinded to this result, the physician who cared for the patient completed another questionnaire. RESULTS: We evaluated 1504 patients (mean age 71.6 years; males 64%; 74% suffering from COPD). Most respondents (93%) used liquid oxygen with mobile device. Fifteen per cent of patients had a prescribed length of oxygen therapy less than 15 h/day; 21% reported to practice oxygen for less than 15 h/day. Patients reported using oxygen for less hours than had been prescribed during the day at rest (P=0.02, k=0.80) during exercise (P=0.002, k=0.72) and at night (P=0.0036, k=0.77). There was no difference between the flow prescribed by the physician and that known and practised by the patient at rest or during sleep; during exercise the flow reported by patients was lower than that prescribed by the physician. Patients used in the night but not at rest or during exercise, a lower level of oxygen flow than what they knew had been prescribed. Fifty-five per cent of patients received indications to modify the oxygen flow in the various situations of life. Liquid oxygen was almost always useful to decrease breathlessness. Most (84%) patients possessed a mobile device, but only 40% declared they used it daily, 'shame' being indicated as the principal barrier. On the physicians' side, we found that the criteria used in prescribing did not always correspond to evidence-based recommendations. CONCLUSION: The widespread use of liquid oxygen did not automatically assure optimal adherence to the prescribed treatment as regards times and modality of oxygen use. A better education of patients, relatives, and the general public, as well as increased self-assessment on the part of health caregivers would improve the practice of LTOT in Italy. PMID- 16242927 TI - Microfluidic-based diagnostics for cervical cancer cells. AB - The use of biomarkers has facilitated the detection of specific tumor cells. However, the technology to apply these markers in a clinical setting has not kept pace with their increasing availability. In this project, we use an antibody based microfluidics platform to recognize and capture cervical cancer cells. Because HPV-16 infection of cervical cells and up-regulation of alpha6-integrin cell surface receptors are correlated, we utilized alpha6-integrin as a capture antibody bound to the channel surface. Normal human glandular epithelial cells (HGEC), human cervical stromal cells (HCSC) and cervical cancer cells (HCCC) were suspended in PBS and flowed through the system. Greater than 30% of the cancer cells were captured while the capture of the normal cell types was less than 5%. The technique is sensitive and accurate. It is potentially useful in the detection of cervical cancer at all stages, as well as other of cancers with similar characteristics of cell surface antigen expression. PMID- 16242928 TI - Prognostic relevance of methylation markers in patients with non-muscle invasive bladder carcinoma. AB - There is increasing evidence for the role of epigenetic gene silencing in superficial bladder cancer. The aim of the current study was to investigate the prognostic value of epigenetic alterations in patients with non-muscle invasive bladder carcinoma. We checked the methylation status of 20 cancer associated genes (p14ARF, p16 CDKN2A, STAT-1, SOCS-1, DR-3, DR-6, PIG-7, BCL-2, H-TERT, BAX, EDNRB, DAPK, RASSF-1A, FADD, TMS-1, E-Cadherin, ICAM-1, TIMP-3, MLH-1, COX-2) for DNA methylation. We analysed microdissected tumour samples from 105 consecutive patients with primary non-muscle invasive bladder carcinoma. Quantitative methylation analysis of CpG sites in the promoter region of the genes was performed with methylation sensitive quantitative real time PCR ('Methylight'). Univariate analysis for association with tumour recurrence was carried out with the Kaplan-Meier analysis and the log-rank test. Follow-up data were available in 95/105 patients (91.4%). A tumour recurrence was observed in 26 patients (27.3%). We could identify six genes (SOCS-1, STAT-1, BCL-2, DAPK, TIMP-3, E-Cadherin), where methylation was associated with tumour recurrence. In Kaplan-Meier analysis, TIMP-3 showed a significant association with recurrence free survival. Methylation of TIMP-3 predicted prolonged disease free interval. In this study, we report a comprehensive analysis on prognostic relevance of gene methylation in non-muscle invasive bladder cancer. We identified one gene (TIMP-3) where methylation was associated with a more favourable outcome. Our data strongly support the usefulness of gene methylation as a prognostic marker in patients with non-muscle invasive bladder cancer. PMID- 16242929 TI - Differential effects of Vitamin D analogs on bone formation and resorption. AB - Deficiency in Vitamin D and its metabolites leads to a failure in bone formation primarily caused by dysfunctional mineralization, suggesting that Vitamin D analogs might stimulate osteoblastic bone formation and mineralization. In this study, we compare the effect of selected Vitamin D analogs and active metabolite, 1alpha,25-dihydroxyvitamin D(3), 19-nor-1alpha, 25-dihydroxyvitamin D(2), and 1alpha-hydroxyvitamin D(2) or 1alpha,25-dihydroxyvitamin D(2) on bone formation and resorption. In a mouse calvariae bone primary organ culture system, all Vitamin D analogs and metabolite tested-stimulated collagen synthesis in a dose dependent manner and 19-nor-1alpha, 25-dihydroxyvitamin D(2) was the most efficacious among three. 19-nor-1alpha, 25-dihydroxyvitamin D(2) and 1alpha,25 dihydroxyvitamin D(2) showed similar potencies and 1alpha,25-dihydroxyvitamin D(3) was less potent than others. Osteocalcin was also up-regulated in a dose dependent manner, suggesting that the three Vitamin D analogs have the equal potencies on bone formation. 25-Hydroxyvitamin D-24-hydroxylase expression was induced in a dose-dependent manner and 19-nor-1alpha, 25-dihydroxyvitamin D(2) was less potent than other two compounds. In a mouse calvariae organ culture, all induced a net calcium release from calvariae in a dose-dependent manner, but the potency is in the order of 1alpha,25-dihydroxyvitamin D(2) congruent with1alpha,25-dihydroxyvitamin D(3)>19-nor-1alpha, 25-dihydroxyvitamin D(2). In a Vitamin D/calcium-restricted rat model, all caused an elevation in serum calcium in a dose-dependent manner. There is no significant difference between 1alpha,25 dihydroxyvitamin D(3) and 1alpha-hydroxyvitamin D(2) in potencies, but 19-nor 1alpha, 25-dihydroxyvitamin D(2) is at least 10-fold less potent than the other two compounds. Our results suggest that Vitamin D analogs have direct effects on bone resorption and formation, and 19-nor-1alpha, 25-dihydroxyvitamin D(2) may be more effective than 1alpha,25-dihydroxyvitamin D(3) and 1alpha-hydroxyvitamin D(2) on stimulating anabolic bone formation. PMID- 16242930 TI - Vitamin D and cancer. Proceedings of the vitamin D workshop meeting. November 17 19, 2004. National Institutes of Health, Bethesda, Maryland, USA. PMID- 16242931 TI - Antibodies and vaccines--hope or illusion? AB - The search for target molecules on tumor cells eliciting strong immune responses in cancer patients has been pursued over decades. Growth factors and their respective receptors were discovered as suitable targets for passive or active immunotherapy approaches. Monoclonal antibodies directed against some of these targets like the proto-oncogene HER2/neu have become an accepted standard of therapy in the clinical management of subgroups of HER2/neu overexpressing breast cancer patients and in other malignancies. Antibodies against multiple other target molecules like epidermal growth factor receptor (EGFR), vascular endothelial growth factor (VEGF), etc., are explored in ongoing trials in order to enter clinical practice in the near future. More recently, potent techniques have been developed to identify cancer antigens eliciting spontaneous immune responses in cancer patients. Cancer vaccination strategies targeting some of these cancer antigens have also been developed, and are maturing for clinical application. With reliable immunomonitoring techniques in place it has been shown that vaccination with some of these cancer antigens may induce strong integrated (humoral and cellular) immune responses in antigen-positive cancer patients. A prominent example is the cancer testis (CT-) antigen NY-ESO-1, which is expressed in 30% of all breast cancers. NY-ESO-1 is one of the most immunogenic human cancer antigens known to date. The aim of ongoing clinical trials is to induce or augment preexisting immune responses in cancer patients with strong NY-ESO-1 positive disease. There is preliminary evidence that patients with strong NY-ESO 1-specific immune responses have more favorable courses of disease. In several clinical phase I trials targeting HER2/neu it was shown that antigen-specific T cell responses could be induced. Another new cancer antigen explored for cancer vaccination is the breast differentiation antigen NY-BR-1, expressed in 70% of all tested primary breast cancers. Although this cancer antigen is still in preclinical testing, its strong and restricted pattern of expression in breast cancer makes it a promising target for clinical development. For all cancer vaccines there is mounting evidence that the stage of disease to be targeted is minimal residual disease or in adjuvant settings. PMID- 16242932 TI - The RGK family: a regulatory tail of small GTP-binding proteins. AB - RGK proteins are small Ras-related GTP-binding proteins that function as potent inhibitors of voltage-dependent calcium channels, and two members of the family, Gem and Rad, modulate Rho-dependent remodeling of the cytoskeleton. Within the Ras superfamily, RGK proteins have distinct structural and regulatory characteristics. It is an open question as to whether RGK proteins catalyze GTP hydrolysis in vivo. Binding of calmodulin and the 14-3-3 protein to RGK proteins controls downstream pathways. Here, we discuss the structural and functional properties of RGK proteins and highlight recent work by Beguin and colleagues addressing the mechanism of Gem regulation by calmodulin and 14-3-3. PMID- 16242933 TI - Chiari malformation and odontoid panus causing craniovertebral stenosis in a child with Crouzon's syndrome. AB - Crouzon's disease is a well-known disorder affecting multiple organ systems, specifically a craniofacial disorder with highly variable penetrance and severity of deformity. Crouzon's patients typically have anomalies of the skull base leading to gross distortion of the cranium and in some cases the cervicocranium. We present a 5-year-old girl with Crouzon's disease who suffered from an acquired Chiari I malformation after insertion of a ventriculoperitoneal shunt and a coexistent ventral odontoid panus. Both these lesions were causing cervicomedullary compression. The literature is controversial on the surgical management of anterior and posterior compression at the craniocervical junction. We review the literature on surgical options for decompression at the craniocervical junction and offer our surgical case as a treatment option for patients in this rare clinical situation. PMID- 16242934 TI - Multi-modality neurophysiological monitoring during surgery for adult tethered cord syndrome. AB - During complex microneurosurgery performed in patients with tethered cord syndrome, the conus medullaris and the roots that innervate the lower limbs, bladder and bowel are potentially exposed to damage. The aim of multimodality intraoperative monitoring (IOM) is to reduce the risk of inadvertent injury of neural tissue. We simultaneously record tibial nerve somatosensory evoked potentials (SSEPs) from the scalp and free run electromyography (EMG) of limb muscles supplied by L2 to S2 roots, anal and urethral sphincters. We also identify critical neural structures in the operative field, including the conus and exiting nerve roots, with a nerve stimulator to evoke EMG. SSEPs assess the sensory pathways mainly mediated by the S1 roots. Continuous EMG provides the surgeon with immediate auditory feedback resulting from irritative discharges triggered by manipulation of nerve fibres. Microstimulation can distinguish the filum terminale, scar tissue and invasive tumors from functional neural tissue, thus minimizing the risk of iatrogenic injury. Overall multimodality IOM proves a valuable adjunct to microneurosurgery of the lumbosacral spine. PMID- 16242935 TI - Foraminal migration of a lumboperitoneal shunt catheter tip. AB - Although lumboperitoneal shunts have some advantages over other shunt types, they are also associated with unique complications, including scoliosis, back pain, and sciatica. We report a case of foraminal migration of a lumboperitoneal shunt catheter tip, which resulted in radicular pain and neurological deficit. PMID- 16242936 TI - Ventriculoperitoneal shunt catheter masquerading as ascariasis. AB - We describe extrusion of a ventriculoperitoneal shunt catheter through the anus occurring in an eight month child with hydrocephalus. This is a rare complication of ventriculoperitoneal shunt. Perforation of the bowel by the peritoneal catheter of the shunt occurred without peritonitis or retrograde infection of the shunt system. Pathogenesis and the literature is reviewed. PMID- 16242937 TI - First case report of X linked dystonia parkinsonism (XDP) or 'lubag' in Australia. AB - PURPOSE: To present the first genetically supported case of X linked dystonia parkinsonism (XDP) or 'lubag' reported in an Australian hospital. METHODS: We performed PCR amplification of microsatellite markers in and around the previously described segregating region for the XDP haplotype. RESULTS: Linkage was confirmed using markers ZNF261, DXS10017, and DXS10018. CONCLUSION: We present the first case of XDP or 'lubag' reported in an Australian hospital. It highlights the enlarging role of genetic testing in facilitating the diagnosis of dystonia in a clinical environment where a disease like XDP is rare, and where a corroborating family history may be unavailable. PMID- 16242938 TI - Observing three-dimensional human microvascular and myogenic architecture using conventional fluorescence microscopy. AB - Microangiography and vascular casting have previously been used to demonstrate the three-dimensional architecture of human uterine microvasculature. However, a limitation of these perfusion-dependent techniques is the difficulty in identifying surrounding tissue components. We have previously shown that it is possible to visualise microvascular networks on the cut surfaces of fresh tissue specimens by diffusive labelling of vascular endothelium with fluorescently conjugated UEA-1 lectin. Unlike perfusion methods that are limited to accessible vascular networks, diffusive fluorescence labelling (DFL) allows additional visualisation of extravascular cellular components, such as smooth muscle. Following UEA-1 DFL, smooth muscle-myosin and -actin were then visualised by immunolocalisation on the acetone-fixed tissue pieces. This allowed clear three dimensional distinction between the vascular and muscle architecture of the myometrium and endometrium. This method can also be applied for studying the relative distribution of microvascular and muscle architecture in leiomyomas (fibroids). The techniques described in this methodological study provide a simple way of directly examining the uterine vasculature in three dimensions using conventional microscopy, while also distinguishing myometrial from endometrial parts of the network. PMID- 16242940 TI - The RACHS-1 risk categories reflect mortality and length of stay in a Danish population of children operated for congenital heart disease. AB - OBJECTIVE: The Risk Adjusted classification for Congenital Heart Surgery (RACHS 1) was created in order to compare in-hospital mortality for groups of children undergoing surgery for congenital heart disease. The method was evaluated with two large multi-institutional data sets-the Paediatric Cardiac Care Consortium (PCCC) and Hospital Discharge (HD) data from three states in the USA. The RACHS-1 classification was later applied to a large German paediatric cardiac surgery population in Bad Oeynhausen (BO), where it was found that the RACHS-1 categories were also associated with length of stay. We applied the RACHS-1 classification to the 957 operations performed during January 1996 to December 2002 at Skejby Sygehus, Denmark and we examined the association between the RACHS-1 categories, in-hospital mortality and length of stay in the Intensive Care Unit. METHODS: The operations were classified according to the six RACHS-1 categories by matching the procedure of each patient with a risk category. The ability of the RACHS-1 classification to predict mortality in our population was examined by estimating the area under the receiver operator characteristic (ROC) curve. Likelihood ratio chi(2) tests were used to compare the distribution of RACHS-1 categories and the distribution of mortality with PCCC, HD and BO. Linear regression was used to examine the correlation between the RACHS-1 categories and length of stay in the Intensive Care Unit. RESULTS: The RACHS-1 category frequencies in our population were: category 1: 18.4%, category 2: 37.4%, category 3: 34.6%, category 4: 8.2%, category 5: 0% and category 6: 1.5%. The overall ability of the RACHS-1 classification to predict in-hospital mortality (area under the ROC curve 0.741; 95% confidence interval=0.690; 0.791) was equal to the findings from larger populations. We found no differences in the category specific mortality when comparing with the larger reported series. There was a positive association between RACHS-1 category and length of stay in the Intensive Care Unit. CONCLUSIONS: The RACHS-1 classification can also be used to predict in-hospital mortality and length of stay in the Intensive Care Unit in a small volume centre. PMID- 16242941 TI - Retrograde pulmonary embolectomy in massive pulmonary embolism. AB - The purpose of this study was introduction and evaluation of efficacy and safety of retrograde thromboembolectomy in acute massive pulmonary emboli. The method is described in a 56-year-old woman with acute massive pulmonary thromboemboli. Postoperative course was uneventful. The described surgical technique is not a panacea and definitely not the whole answer, but is a big part of the solution and may be accompanied with less adverse effects. Additionally, there is a need of being reviewed further in large experimental studies and measurements before it could be used safely as a new technique. PMID- 16242942 TI - Adjuvant chemotherapy in patients 80-89 years of age with non-small cell lung cancer. PMID- 16242943 TI - Quality assessment of distal S(2)AS connector anastomosis by 13 MHz epicardial ultrasound. AB - OBJECTIVE: During application of a distal coronary bypass connector, we employed 13 MHz epicardial ultrasound to evaluate quantitative caliper measurements for vessel size matching and to assess anastomosis quality after connector deployment. METHODS: Two S(2)AS connector anastomoses were constructed on ex vivo pressure-perfused porcine hearts. Epicardial ultrasound measurements of the connector ring and anastomosis were compared to intravascular ultrasound measurement and cast dimensions. In 21 pigs, anastomotic sites with internal diameter of 2.25-3.0mm (internal mammary artery, IMA) and 1.8-2.2mm (left anterior descending coronary artery, LAD) were selected using external caliper and epicardial ultrasound measurements. Anastomoses were visualized and assessed intraoperatively (beating heart, n=21) and at 3 and 6 months' follow-up (explanted heart, n=10 each). RESULTS: Epicardial ultrasound underestimated connector dimension by < or =5% versus intravascular ultrasound and deviated < or =13% from cast dimensions for other anastomotic measurements. Caliper estimates of internal IMA and LAD diameter differed from ultrasound by -3+/-6% and -2+/-7% (mean+/-SD), respectively. Intraoperatively, the anastomotic orifice was flawless in all animals. It remained fully patent at 3 and 6 months by ultrasound, which was confirmed by histology. The connector to LAD percentage diameter stenosis changed from -12+/-5% intraoperatively to -1+/-7% at 3 months and from -5+/-6% intraoperatively to -16+/-13% at 6 months, in the growing pig model. CONCLUSIONS: In the pig, external caliper measurements provided a reliable quantitative estimate of inner graft and coronary diameter for connector size matching. Epicardial 13 MHz ultrasound is a promising method to assess coronary anastomosis quality even when connector metal is present. PMID- 16242944 TI - Synchronously counterpulsating extracorporeal life support enhances myocardial working conditions regardless of systemic perfusion pressure. AB - OBJECTIVE: A new pulsatile extracorporeal life support (pECLS) system has entered the market. We wanted to investigate what potential advantages pECLS may have over current non-pulsatile systems (NPS). Our research was focused on the pump's functional interaction with the left ventricle and the coronary circulation. METHODS: Extensive hemodynamic measurements were performed during asynchronous and synchronous pECLS in 10 calves. The two extremes regarding LV afterload, namely systolic arrival (SA) and diastolic arrival (DA) of the pump pulse were studied. RESULTS: SA was associated with increased oxygen consumption (+57%) and decreased diastolic coronary perfusion (-43%). DA increased left ventricular output (DA: 4.5+/-2.4 l/min vs SA: 3.5+/-2.2 l/min), LV ejection fraction (+10%), and ventricular efficiency (+17%). Mean aortic pressure and mean coronary flow were only marginally affected by pulse incidence. Systolic impairment was more pronounced with higher bypass flows. These results indicate that myocardial working conditions can be optimized by phasing pECLS ejection into cardiac diastole. CONCLUSION: We conclude that during pECLS, myocardial working conditions can be improved by avoidance of systolic impairment. Synchronously counterpulsating pECLS could be a more economic and versatile alternative to NPS or NPS combined with intra-aortic balloon pumping. The potential benefits of synchronously counterpulsating pECLS over the current alternatives remain to be investigated. PMID- 16242946 TI - Stem cell therapy for myocardial regeneration: creating hype ignoring reality. PMID- 16242947 TI - Emergency endovascular repair for traumatic injury of the inferior vena cava. AB - Conventional open repair of the traumatic injuries of the inferior vena cava still represents a surgical challenge, since it carries high morbidity and mortality rates close to 100% in emergency setting. Surgical techniques required the total mobilization of the liver in the former case and the anatomic exposure of the aorto-iliac bifurcation in the latter. We report a case of uncontrolled hemorrhage from the inferior vena cava, managed by endovascular technique. A 65 year-old woman underwent endovascular stent-grafting for traumatic injury of the inferior vena cava. We performed the endovascular approach as we similarly did for aortic repair using a simple groin access through the common femoral vein; no additional dissection of the inferior vena cava or adjacent structures was needed using endovascular approach, thus avoiding additional operative trauma, and time with the stent-graft procedure was 9 min. Inferior vena cava stent-grafting without laparotomy and retroperitoneal dissection is feasible and could be a very attractive alternative to open surgery for rupture of the vena cava. PMID- 16242948 TI - Inhaled iloprost to control residual pulmonary hypertension following pulmonary endarterectomy. AB - OBJECTIVE: Pulmonary endarterectomy (PEA) is the standard therapy for patients with chronic thromboembolic pulmonary hypertension (CTEPH). In the immediate postoperative period, persistent pulmonary hypertension increases the risk of acute respiratory or right heart failure. In pulmonary arterial hypertension, prostanoid inhalation has been found to improve pulmonary hemodynamics, right ventricular function, gas exchange, and clinical outcome. We report the results of a double-blinded randomized trial with the aerosolized prostacyclin analogue iloprost in patients with residual pulmonary hypertension after PEA. METHODS: Twenty-two patients (age, 55+/-13 years; 8 females; propofol- and sufentanil based anesthesia; pressure-controlled mechanical ventilation) were randomized to receive either a single dose of 25 microg aerosolized iloprost (iloprost group; n=11) or normal saline (placebo group; n=11) immediately after postoperative ICU admission. Primary endpoints were changes in gas exchange, pulmonary and systemic hemodynamics, and clinical outcome. RESULTS: Iloprost significantly enhanced cardiac index (CI) and reduced mean pulmonary arterial pressure (mPAP) and pulmonary vascular resistance [PVR (dynscm(-5))] in contrast to placebo. Placebo: pre-inhalation 413+/-195 versus post-inhalation 404+/-196 at 30 min (p=0.051), 415+/-189 at 90 min (p=0.929). Iloprost: pre-inhalation 503+/-238 versus post inhalation 328+/-215 at 30 min (p=0.001), 353+/-156 at 90 min (p=0.003). Blood oxygenation remained unchanged. CONCLUSION: In addition to the effect of PEA, iloprost reduces residual postoperative pulmonary hypertension, decreases right ventricular afterload and may facilitate the early postoperative management after PEA. PMID- 16242949 TI - Detailed preoperative information--cruelty or improving quality of informed consent. PMID- 16242950 TI - Coexistence of left pulmonary artery sling and aortopulmonary window complicated with difficult airway-a rare congenital cardiopulmonary defect. AB - Both the pulmonary artery sling and the aortopulmonary window are unusual and serious anomalies. In patients with sling the airway might be compromised by associated cartilaginous o-rings in variant segments. In aortopulmonary window the clinical presentations are similar to the large patent ductus arteriosus or ventricular septal defect, but the surgical procedure is quite difficult and different. We operated an infant with a rarely seen cardiac defect, the coexistence of left pulmonary artery sling and Type 3 aortopulmonary window, when he was 35 days old. Postoperative repeated bronchospasm caused prolonged ventilation and hospitalization. Patient was extubated 28 days later and discharged at the age of 80 days without any events. Besides, the reconstructed three-dimensional images of ultrafast computed tomography offered us an interesting view other than surgical findings. A case of two such rare lesions coexisting was not to be found in the literature review, so we report this case because of its rarity and clinical interest. PMID- 16242951 TI - Myocardial protection with intermittent cross-clamp fibrillation: does preconditioning play a role? AB - OBJECTIVE: Previously, we showed intermittent cross-clamp fibrillation afforded equivalent protection to cardioplegia. This study examined whether protection induced by intermittent cross-clamp fibrillation involves an ischemic preconditioning mechanism. METHODS: Isolated Langendorff-perfused rat hearts were subjected to three different studies to determine: Study 1, whether a single intermittent cross-clamp fibrillation episode (10 min) and reperfusion (10 min) before prolonged ischemia acts as a preconditioning trigger for protection; Study 2, whether cardioprotection induced by intermittent cross-clamp fibrillation alone (no prolonged ischemia) involves a preconditioning mechanism; Study 3, whether intermittent cross-clamp fibrillation cardioprotection can be prevented by targeting putative components of the preconditioning mechanism (protein kinase C or the mitochondrial ATP-sensitive potassium (K(ATP)) channel). Hearts were reperfused (60 min) and recovery of function (left ventricular developed pressure measured using an intraventricular balloon) and myocardial injury (creatine kinase leakage) were measured. RESULTS: In Study 1, recovery of function in the single intermittent cross-clamp fibrillation hearts was 61+/-3% (mean+/-SEM) (p<0.05) compared to 41+/-2% in control group; glibenclamide (a non-specific ATP sensitive potassium (K(ATP))-channel blocker) prevented this preconditioning protection (37+/-4%). In Study 2, recovery of function in intermittent cross clamp fibrillation hearts (62+/-3%) was significantly (p<0.05) higher than intermittent cross-clamp fibrillation hearts treated with glibenclamide (33+/-2%) and ischemia hearts (30+/-5%). In Study 3, protection by intermittent cross-clamp fibrillation (60+/-3%; p<0.05) was attenuated by protein kinase C inhibition (chelerythrine, 34+/-3%) and mitochondrial K(ATP)-channel blockade (5 hydroxydecanoate, 27+/-4%) to levels not significantly different from that of ischemia hearts (25+/-4%). CONCLUSIONS: The cardioprotective efficacy of intermittent cross-clamp fibrillation was attenuated by protein kinase C inhibition or K(ATP)-channel blockade. Involvement of these putative preconditioning cascade components in association with cardioprotection induced by intermittent cross-clamp fibrillation, suggests a role for the ischemic preconditioning mechanism. PMID- 16242952 TI - Pharmaceutical analysis of 5-aminolevulinic acid in solution and in tissues. AB - Quantification of 5-aminolevulinic acid (ALA) in solution, and methods used to achieve this, have been extensively reported in the literature. However, validated methods have only rarely been presented and never have methods been compared. Due to a necessity in drug delivery research for optimised and validated methods for determination of ALA in solution, this paper compares, for the first time, two such methods validated to International Conference on Harmonisation (ICH) standards. Of major importance, derivatisation of ALA with acetyl acetone and formaldehyde was found to be more suitable for routine fluorimetric HPLC analysis of ALA than derivatisation with o-phthaldialdehyde and 2-mercaptoethanol. This former method was successfully utilised in the comparison of in vitro drug release from a proprietary ALA cream and a novel bioadhesive patch-based system. In addition, determination of ALA in tissue is necessary to compare different topical formulations, in terms of their ability to deliver the drug successfully, and different tissue types, to assess their barrier properties to penetration of the drug. Consequently, this paper also describes the use of liquid scintillation spectroscopy as an analytical tool for rapid, convenient and routine quantification of ALA in tissue and determination of penetration depth following topical application of creams and patches. PMID- 16242953 TI - Manipulating the fragmentation patterns of phosphopeptides via gas-phase boron derivatization: determining phosphorylation sites in peptides with multiple serines. AB - Trivalent boron species readily react with protonated phosphopeptides to give addition products with the loss of boron ligands. In the present study, trimethoxyborane (TMB), diisopropoxymethylborane (DIPM), and diethylmethoxyborane (DEMB) were allowed to react with four phosphopeptides, VsSF, LSsF, LsGASA, and VSGAsA (lower-case s indicates phosphoserine). Each of the phosphopeptides contains one serine that is phosphorylated and one that is not. Under collision activated dissociation (CAD) conditions, the boron-derivatized peptides give fragmentation patterns that differ significantly from that of the protonated phosphopeptide. The patterns vary, depending on the number of labile (i.e., alkoxy) ligands on the boron. In general, boron derivatization increases the yield of phosphate-containing sequence ions, but dramatic effects are only seen with certain reagent/peptide combinations. However, the suite of reagents provides a means of altering and increasing the information content of phosphopeptide CAD spectra. PMID- 16242954 TI - Stability of the homopentameric B subunits of shiga toxins 1 and 2 in solution and the gas phase as revealed by nanoelectrospray fourier transform ion cyclotron resonance mass spectrometry. AB - The assembly of the B subunits of Shiga toxins (Stx) 1 and 2 and the influence of solution conditions (protein concentration, temperature, pH, and ionic strength) on it are investigated using temperature-controlled nanoflow electrospray (nano ES) ionization and Fourier-transform ion cyclotron resonance mass spectrometry. Despite the similar higher order structure predicted by X-ray crystallography analysis, the B(5) homopentamers of Stx1 and Stx2 exhibit differences in stability under the solution conditions investigated. At solution temperatures ranging from 0 to 60 degrees C and subunit concentrations ranging from 5 to 85 microM, the Stx1 B subunit exists almost entirely as the homopentamer in aqueous solutions, independent of the ionic strength. In contrast, the degree of assembly of Stx2 B subunit is strongly dependent on temperature, subunit concentration, and ionic strength. At subunit concentrations of more than 50 microM, the Stx2 B subunit exists predominantly as a pentamer, although smaller multimers (dimer, trimer, and tetramer) are also evident. At lower concentrations, the Stx2 B subunit exists predominantly as monomer and dimer. The relative abundance of multimeric species of the Stx2 B subunit was insensitive to the ion source conditions, suggesting that gas-phase dissociation of the pentamer ions in the source does not influence the mass spectrum. Blackbody infrared radiative dissociation of the protonated B(5) ions of Stx2 at the +12 and +13 charge states proceeds, at reaction temperatures of 120 to 180 degrees C, predominantly by the ejection of a single subunit from the complex. Dissociation into dimer and trimer ions constitutes a minor pathway. It follows that the dimer and trimer ions and, likely, the monomer ions observed in the nano-ES mass spectra of Stx2 B subunit originated in solution and not from gas-phase reactions. It is concluded that, under the solution conditions investigated, the homopentamer of Stx2 B subunit is thermodynamically less stable than that of Stx1 B subunit. Arrhenius activation parameters determined for the protonated Stx2 B(5) ions at the +12 and +13 charge states were compared with values reported for the corresponding B(5) ions of Stx1 B subunit. In contrast to the differential stability of the Stx1 and Stx2 B pentamers in solution, the dissociation activation energies (E(a)) determined for the gaseous complexes are indistinguishable at a given charge state. The similarity in the E(a) values suggests that the protonated pentamer ions of both toxins are stabilized by similar intersubunit interactions in the gas phase, a result that is in agreement with the X-ray crystal structures of the holotoxins. PMID- 16242955 TI - Cloning, expression, purification, and characterization of zebrafish cytosolic serine hydroxymethyltransferase. AB - A cDNA which encodes for zebrafish serine hydroxymethyltransferase (SHMT) has been cloned into a pET43.1a vector as a NdeI-EcoRI insert and transformed into HMS174(DE3) cells. After induction with isopropyl thiogalactoside, the enzyme was purified with a three-step purification protocol and about 15 mg of pure enzyme was obtained per liter of culture. Spectral and structural characteristics of the recombinant zebrafish SHMT are similar to the rabbit and human cytosolic SHMT. Kinetic constants for the natural substrates l-serine and tetrahydrofolate are also comparable to the values obtained previously for the rabbit and human cytosolic enzyme. PMID- 16242957 TI - Prion protein from Xenopus laevis: overexpression in Escherichia coli of the His tagged protein and production of polyclonal antibodies. AB - Prion protein (PrP) and PrP-related proteins have been identified in reptiles, amphibians, and fishes by means of cDNA cloning, genome database searching and comparative genomics. However, no studies have been reported so far on the expression of PrP at the protein level in those animals. This report presents a procedure to obtain and purify recombinant PrP from Xenopus laevis expressed in Escherichia coli as a fusion protein in which mature PrP (residues 21-194) is linked to a 35-amino acid N-terminal extension containing a hexahistidine stretch. The protein was used to raise and purify by affinity chromatography anti Xenopus PrP polyclonal antibodies which were suitable to detect the presence of PrP in Xenopus brain by Western blot. This is the first report of a positive identification of PrP in amphibian at the protein level. Anti-Xenopus PrP antibodies do not cross react with PrP from different sources (human, bovine, sheep, and turtle). Similarly, Xenopus PrP do not react with anti-turtle PrP(143 248) antibodies. PMID- 16242956 TI - High yield synthesis and characterization of phosphorylated recombinant human procathepsin D expressed in mammalian cells. AB - We used a vaccinia virus expression system for the production of recombinant human cathepsin D (CD), a lysosomal protease implicated in various patho physiological processes including cancer, neurodegeneration, and development. The recombinant protein was successfully expressed in various human and non-human cells. It was correctly synthesized as a glycosylated 53 kDa precursor (proCDrec) that reacted with a polyclonal antibody against residues 7-21 of the propeptide sequence. In contrast to the control, in cells infected with the recombinant virus proCDrec was largely secreted into the culture medium, although it contained high-mannose oligosaccharides with uncovered mannose-6-phosphate residues. Intracellular proCDrec was processed into the 48 kDa intermediate single-chain and the 31 plus 13 kDa double-chain forms, however, the processing was slower than in normal cells. A method based on Pepstatin A-affinity chromatography allowed to isolate the recombinant protein from the medium of infected cells. Based on its latency in activity assay at acid pH and on its reactivity with antibodies specific for the N-terminus, the purified protein was judged to be in the inactive precursor form. During incubation at acid pH the purified proCDrec underwent autocatalytic processing and acquired pepstatin A sensitive enzyme activity, as expected for correctly folded proCD. Antiserum raised in rabbits against proCDrec specifically reacted with human, but not with mouse proCD under non-denaturing conditions. We conclude that our vaccinia virus directed proCDrec displays structural and functional features resembling those of native human proCD. This system can therefore be exploited for the synthesis of large quantities of human proCD, allowing further studies on the structure and function of this interesting protein. PMID- 16242958 TI - Using cost and health impacts to prioritize the targeted testing of tuberculosis in the United States. AB - PURPOSE: Evaluation improves efficiency and effectiveness. Current U.S. tuberculosis (TB) control policies emphasize the treatment of latent TB infection (LTBI). However, this policy, if not targeted, may be inefficient. We determined the efficiency of a state-law mandated TB screening program and a non state-law mandated one in terms of cost, morbidity, treatment, and disease averted. METHODS: We evaluated two publicly funded metropolitan TB prevention and control programs through retrospective analyses and modeling. Main outcomes measured were TB incidence and prevalence, TB cases averted, and cost. RESULTS: A non state-law mandated TB program for homeless persons in Tarrant County screened 4.5 persons to identify one with LTBI and 82 persons to identify one with TB. A state-law mandated TB program for jail inmates screened 109 persons to identify one with LTBI and 3274 persons to identify one with TB. The number of patients with LTBI treated to prevent one TB case was 12.1 and 15.3 for the homeless and jail inmate TB programs, respectively. Treatment of LTBI by the homeless and jail inmate TB screening programs will avert 11.9 and 7.9 TB cases at a cost of 14,350 US dollars and 34,761 US dollars per TB case, respectively. CONCLUSIONS: Mandated TB screening programs should be risk-based, not population-based. Non mandated targeted testing for TB in congregate settings for the homeless was more efficient than state-law mandated targeted testing for TB among jailed inmates. PMID- 16242959 TI - Why do postmenopausal African-American women not benefit from overall breast cancer mortality decline? AB - BACKGROUND: An overall decline in U.S. female breast cancer mortality in the 1990s has been reported. However, several studies have shown that mortality trends are different for White women and African-American (AA) women. The purpose of this study was to assess differences in time trends and patterns of female breast cancer mortality among women by race and age in Ohio. MATERIAL AND METHODS: Joinpoint regression (JR) and age-period-cohort (APC) approaches were used to evaluate temporal changes in mortality and to assess period and birth generation impacts on observed patterns. Logistic regression was used to assess racial differences in tumor staging and grading among women diagnosed with breast cancer in Ohio from 1996 to 2000. Mortality data were obtained from NCHS (National Center for Health Statistics) via Surveillance Research Program, National Cancer Institute SEER*Stat software; Ohio incidence data were provided by the Ohio Cancer Incidence Surveillance System. RESULTS: Among women aged 30 74, a significant decline of 2.8% was noted since 1988 for White women. AA women in this age group have experienced significant decline (by 0.9%) since 1983. White women aged 30-39 years experienced a decline in mortality of 3.5% per year in the period 1986-2001, while decline by 2.3% was observed among Black women of that age since 1984. Among the age categories 40-49, 50-59, and 60-74, a decline in mortality rates was observed among White women in the 1990s. The decline was observed also among AA women aged 40-49, beginning in the mid 1980s, but not in the older AA age groups. Specifically, in AA women 60-74 and 75+, a mortality increase was observed within the entire study period (0.9% and 1.4%, respectively). CONCLUSIONS: Analysis of the data for Ohio suggests that AA women do not equally benefit from the overall decline in breast cancer mortality that is often sited. This is especially true for AA postmenopausal women who continue to experience an increase in breast cancer mortality. In light of existing literature and this analysis of data from the state of Ohio, we conclude that the reason for these differences lies mostly in disparities in access to care, as well as in differences in stage at diagnosis and biological determinants (grading) between White and Black women. PMID- 16242960 TI - Prevalence of childhood asthma in urban communities: the impact of ethnicity and income. AB - PURPOSE: The goal of this study was to assess the relationship between hospitalization rates and asthma prevalence in New York City children and investigate the role that sociodemographic factors play in asthma. METHODS: A parent-report questionnaire was distributed in 26 randomly selected New York City public elementary schools, stratified according to neighborhood hospitalization rates. RESULTS: The overall student response rate was 76.9% (5250 students). Prevalence of current asthma was 17.9%, 9.59%, 6.39% (p < .001) in areas of high, median, and low asthma hospitalization rates, respectively. The overall prevalence of current asthma was 13.0%. Children living in predominantly low socioeconomic status (SES) communities had a 70% greater risk of current asthma, independent of their own ethnicity and income level. Asthma prevalence within different ethnic and income groups was consistently lower in neighborhoods of greater socioeconomic status, except among Puerto Rican children, who had high asthma prevalence, regardless of school attended or income. CONCLUSIONS: Prevalence of current asthma is strongly associated with attending a school in a low-income neighborhood. Although hospitalization rates have been used as an indicator of the burden of asthma in a neighborhood, it alone does not reflect the level of disparities that exist among communities with different socioeconomic backgrounds. PMID- 16242961 TI - Mode of delivery is associated with asthma and allergy occurrences in children. AB - PURPOSE: A growing body of evidence indicates that perinatal factors modulate immune development and thereby may affect childhood asthma risk. In this study, we examined the associations between birth by cesarean section (C-section) and atopic disease occurrence in childhood. METHODS: Subjects were born in California between 1975 and 1987 and were 8 to 17 years old during their enrollment in the Children's Health Study. Our analysis was restricted to 3464 children born at or after 37 weeks of gestation with a birth weight of 2500 g or greater based on birth certificate data. Information about sociodemographic factors, reported physician-diagnosed asthma, and other atopic diseases was obtained by using a self-administered structured questionnaire. Logistic regression models were fitted to compute odds ratios (ORs) and 95% confidence intervals (CIs). RESULTS: Children born by C-section were at increased risk for asthma (OR, 1.33; 95% CI, 1.01-1.75), hay fever (OR, 1.57; 95% CI, 1.24-1.99), and allergy (OR, 1.26; 95% CI, 1.03-1.53) compared with those born vaginally. Risk associated with C-section was the same for children regardless of family history of asthma or allergy. CONCLUSION: We conclude that birth by C-section or processes associated with it may increase the risk for atopic disease in childhood. PMID- 16242962 TI - Do cases and controls matched on the first eight digits of their telephone number share geographic proximity and socioeconomic characteristics? AB - PURPOSE: The aim of this study is to assess the geographic proximity of cases and controls who share the same "stem" of their telephone number (area code, prefix, and next two digits) and compare their socioeconomic characteristics. METHODS: We compared residential proximity and characteristics of case-control sets selected in this way with sets consisting of the same cases matched to different controls on all variables except telephone number stem. RESULTS: Mean distance between telephone stem-matched pairs was much less than that between pairs not matched by telephone number stem (3.53 and 20.14 km, respectively). There was slight agreement between cases and controls for some measures of socioeconomic status in both sets. We also compared distances between residences of control respondents who shared the same telephone number stem with those between residences of respondents who shared only the same telephone area code and prefix. Mean distances for the two groups were similar. CONCLUSIONS: Geographic proximity between cases and controls can be achieved by matching on either telephone number stem or only telephone area code and prefix. Adjustment for socioeconomic factors may be needed even if cases and controls live close to each other. PMID- 16242963 TI - Crystal structure of TM1457 from Thermotoga maritima. AB - The crystal structure of a hypothetical protein, TM1457, from Thermotoga maritima has been determined at 2.0A resolution. TM1457 belongs to the DUF464 family (57 members) for which there is no known function. The structure shows that it is composed of two helices in contact with one side of a five-stranded beta-sheet. Two identical monomers form a pseudo-dimer in the asymmetric unit. There is a large cleft between the first alpha-helix and the second beta-strand. This cleft may be functionally important, since the two highly conserved motifs, GHA and VCAXV(S/T), are located around the cleft. A structural comparison of TM1457 with known protein structures shows the best hit with another hypothetical protein, Ybl001C from Saccharomyces cerevisiae, though they share low structural similarity. Therefore, TM1457 still retains a unique topology and reveals a novel fold. PMID- 16242964 TI - Neutrophils and B-cells in blood and head kidney of Atlantic salmon (Salmo salar L.) challenged with infectious pancreatic necrosis virus (IPNV). AB - Salmon B-cells and neutrophils were studied by flow cytometry in IPNV infected salmon. A highly virulent strain of IPNV was used for challenge of parr and post smolts. The parr were challenged by intraperitoneal (ip) injection while salmon post-smolts were challenged by ip injection or cohabitation. No mortality occurred in the parr groups, but a cumulative mortality of about 50% was obtained in cohabitant infected post-smolt groups and less than 10% in ip challenged post smolts. The virus levels were low in head kidney (HK) samples from survivors compared to dead fish. The percentages of neutrophilic granulocytes and Ig+ cells (B-cells) were analysed using HK and blood samples from survivors. The cell populations were identified by monoclonal antibodies (MAb) E3D9, recognising neutrophils, and G2H3 recognising Ig+ cells (B-cells). Parr sampling for leucocyte analyses took place about 1.5 weeks prior to and about 4 weeks post challenge. This corresponded to about 8 and 2.5 weeks before the fish were adapted to seawater transfer. In parr head kidney leucocytes (HKL) we observed significantly lower (p < 0.05) levels of neutrophils in ip infected fish compared to non-infected control fish. The post-smolt sampling from infected fish took place 2 weeks prior to and in the fifth and sixth week post challenge. HKL samples from both surviving cohabitants and ip injected fish had significantly (p < 0.05) lower levels of neutrophils than non-infected control fish. The cohabitant fish also had significantly (p < 0.05) higher levels of B-cells in HKL compared to ip injected fish. No significant changes in B-cells in HKL or peripheral blood leucocytes (PBL) was observed in infected parr or ip infected post-smolts compared to control fish. The relative leucocyte levels of the fish prior to challenge and in non-infected control fish are in accordance with earlier findings. The results indicate that non-specific immune cells like neutrophils are highly influenced by IPNV infection of parr and post-smolts several weeks post challenge. PMID- 16242965 TI - Brain electrical tomography (BET) analysis of induced gamma band responses during a simple object recognition task. AB - The formation of cortical object representations requires the activation of cell assemblies, correlated by induced oscillatory bursts above 20 Hz (gamma band), which are characterized by trial-by-trial latency fluctuations around a mean of approximately 300 ms after stimulus onset. The present electroencephalogram (EEG) study was intended to uncover to the generators of induced gamma band responses (GBRs) and to analyze phase-synchronization between these sources. A standard object recognition task was used to elicit gamma activity. At the scalp surface (electrode space), we found an augmentation of induced GBRs after the presentation of meaningful (familiar) as opposed to meaningless (unfamiliar) stimuli, which was accompanied by a dense pattern of significant phase-locking values between distant recording sites. Subsequently, intracranial current density distributions compatible with the observed scalp voltage topographies were estimated by means of VARETA (Variable Resolution Electromagnetic Tomography). In source space brain electrical tomographies (BETs) revealed widespread generators of induced GBRs at temporal, parietal, posterior, and frontal areas. Phase-locking analysis was calculated between re-constructed electrode signals based on separate forward solutions of the observed generators, thereby eliminating the possibly confounding influence of activity from areas not under observation. The results support the view that induced GBRs signify synchronous neuronal activity in a broadly distributed network during object recognition. The localization of the generators of event-related potentials (ERPs), evoked gamma activity, and induced alpha activity revealed different sources as compared to the induced GBR and, thus, seem to mirror complementary functions during the present task as compared to induced high-frequency brain dynamics. PMID- 16242966 TI - Correction for direction-dependent distortions in diffusion tensor imaging using matched magnetic field maps. AB - Diffusion tensor imaging (DTI) has seen increased usage in clinical and basic science research in the past decade. By assessing the water diffusion anisotropy within biological tissues, e.g. brain, researchers can infer different fiber structures important for neural pathways. A typical DTI data set contains at least one base image and six diffusion-weighted images along non-collinear encoding directions. The resultant images can then be combined to derive the three principal axes of the diffusion tensor and their respective cross terms, which can in turn be used to compute fractional anisotropy (FA) maps, apparent diffusion coefficient (ADC) maps, and to construct axonal fibers. The above operations all assume that DTI images along different diffusion-weighting directions for the same brain register to each other without spatial distortions. This assumption is generally false, as the large diffusion-weighting gradients would usually induce eddy currents to generate diffusion-weighting direction dependent field gradients, leading to mis-registration within the DTI data set. Traditional methods for correcting magnetic field-induced distortions do not usually take into account these direction-dependent eddy currents unique for DTI, and they are usually time-consuming because multiple phase images need to be acquired. In this report, we describe our theory and implementation of an efficient and effective method to correct for the main field and eddy current induced direction-dependent distortions for DTI images under a unified framework to facilitate the daily practice of DTI acquisitions. PMID- 16242969 TI - Phylogeny of pholcid spiders (Araneae: Pholcidae): combined analysis using morphology and molecules. AB - The spider family Pholcidae comprises a large number of mainly tropical, web weaving spiders, and is among the most diverse and dominant spider groups in the world. The phylogeny of this family has so far been investigated exclusively using morphological data. Here, we present the first molecular data for the family analyzed in a phylogenetic context. Four different gene regions (12S rRNA, 16S rRNA, cytochrome c oxidase subunit I, 28S rRNA) and 45 morphological characters were scored for 31 pholcid and three outgroup taxa. The data were analyzed both for individual genes, combined molecular data, and molecular plus morphological data, using parsimony, maximum likelihood, and Bayesian methods. Some of the phylogenetic hypotheses obtained previously using morphology alone were also supported by our results, like the monophyly of pholcines and of the New World clade. On the other hand, some of the previous hypotheses could be discarded with some confidence (monophyly of holocnemines, the position of Priscula), and still others need further investigation (the position of holocnemines, ninetines, and Metagonia). The data obtained provide an excellent basis for future investigations of phylogenetic patterns both within the family and among spider families. PMID- 16242968 TI - Optic nerve diffusion tensor imaging in optic neuritis. AB - Diffusion tensor magnetic resonance imaging (DT-MRI) provides in vivo information about the pathology of multiple sclerosis lesions. Increases in mean diffusivity (MD) and reductions in fractional anisotropy (FA) have been found and may represent axonal disruption. The optic nerve is an ideal structure for study by DT-MRI but previous clinical studies did not obtain the full diffusion tensor necessary to calculate MD and FA. In this study, a technique that was specifically developed to achieve full diffusion tensor measurements from the optic nerve (zonal oblique multislice (ZOOM) echoplanar imaging) was applied to 25 patients with a single unilateral episode of optic neuritis at least one year previously, and 15 controls. The intraorbital nerves were segmented on non diffusion-weighted images and the regions of interest transferred to MD, FA, and eigenvalue maps to obtain quantitative data. Quantitative visual testing and electrophysiology were also performed. In affected nerves, mean MD and mean orthogonal eigenvalue lambda( perpendicular) were elevated, and mean FA reduced compared with clinically unaffected contralateral nerves (P < 0.001) and control nerves (P < 0.001). The mean principal eigenvalue lambda?? was significantly increased in affected nerves compared to contralateral unaffected nerves (P = 0.04) but not compared to control nerves (P = 0.13). There was no association of clinical measures of visual function in affected eyes with the DT-MRI parameters but there was a significant correlation of the whole field visual evoked potential (VEP) amplitude with MD (r = -0.57, P = 0.006) and lambda( perpendicular) (r = -0.56, P = 0.007). These findings suggest that optic nerve DT MRI measures provide an indication of the structural integrity of axons. PMID- 16242967 TI - Dynamic physiological modeling for functional diffuse optical tomography. AB - Diffuse optical tomography (DOT) is a noninvasive imaging technology that is sensitive to local concentration changes in oxy- and deoxyhemoglobin. When applied to functional neuroimaging, DOT measures hemodynamics in the scalp and brain that reflect competing metabolic demands and cardiovascular dynamics. The diffuse nature of near-infrared photon migration in tissue and the multitude of physiological systems that affect hemodynamics motivate the use of anatomical and physiological models to improve estimates of the functional hemodynamic response. In this paper, we present a linear state-space model for DOT analysis that models the physiological fluctuations present in the data with either static or dynamic estimation. We demonstrate the approach by using auxiliary measurements of blood pressure variability and heart rate variability as inputs to model the background physiology in DOT data. We evaluate the improvements accorded by modeling this physiology on ten human subjects with simulated functional hemodynamic responses added to the baseline physiology. Adding physiological modeling with a static estimator significantly improved estimates of the simulated functional response, and further significant improvements were achieved with a dynamic Kalman filter estimator (paired t tests, n=10, P<0.05). These results suggest that physiological modeling can improve DOT analysis. The further improvement with the Kalman filter encourages continued research into dynamic linear modeling of the physiology present in DOT. Cardiovascular dynamics also affect the blood-oxygen dependent (BOLD) signal in functional magnetic resonance imaging (fMRI). This state-space approach to DOT analysis could be extended to BOLD fMRI analysis, multimodal studies and real-time analysis. PMID- 16242970 TI - Resection extent versus postoperative outcomes of seizure and memory in mesial temporal lobe epilepsy. AB - OBJECTIVES: To investigate the effects of the resection of hippocampus and temporal neocortex on postsurgical seizure and memory outcomes in mesial temporal lobe epilepsy (mTLE) patients. METHODS: Sixty-eight mTLE patients underwent pre- and postsurgical brain magnetic resonance imaging (MRI). The patients were divided into seizure-free group (SF, N=54) and non-seizure-free group (NSF, N=14). The resection length of hippocampus was determined by the difference between presurgical and postsurgical hippocampus lengths in MRIs. The lengths of resected temporal gyri were measured on three-dimensional MRI reconstruction. Among SF group, 37 patients performed pre- and postsurgical neuropsychological tests. The postsurgical memory decline (PMD) was calculated by subtracting postsurgical memory score from presurgical one in verbal and visual memory tests. RESULTS: The resection length of hippocampus in SF was significantly longer than in NSF (32.7 +/- 7.7 mm versus 25.1 +/- 7.3 mm, t-test, p=0.002), regardless of intersubject difference in the extent of hippocampal sclerosis (logistic regression, p=0.003) while the resection lengths of the lateral temporal gyri were not different between SF and NSF. Overall postsurgical change of verbal or visual memory was not significant. However, regression analysis showed a significant correlation between the resection length of inferior or basal temporal gyrus and verbal PMD (p<0.001) in left TLE patients with seizure-free outcome. CONCLUSION: More resection of hippocampus may predict a better postsurgical seizure-free outcome. The larger resection of inferior or basal temporal gyrus seems to be related to a postsurgical verbal memory decline in left TLE patients. PMID- 16242971 TI - Distal femoral intercondylar notch dimensions and their relationship to composition and metabolism of the canine anterior cruciate ligament. AB - OBJECTIVE: To determine the relationship between the dimensions of the distal femoral intercondylar notch (ICN) and the composition and metabolism of the anterior cruciate ligament (ACL) in three dog breeds with different relevant risks to ligament rupture and subsequent osteoarthritis (OA). DESIGN: ICN measurements were obtained from the femurs of Golden Retrievers (high risk), Labrador Retrievers (high risk) and Greyhounds (low risk). Femoral condyle width and height, ICN height and width indices, and notch shape index were measured using Vernier callipers in all dogs. Intact ACLs were obtained from the same dog breeds for a study of the impinged areas and were analysed for collagen content, collagen cross-links, and sulphated glycosaminoglycan (GAG) content, matrix metalloproteinase (MMP)-2 and the tissue inhibitors of metalloproteinases (TIMPs) 1 and -2. RESULTS: Femoral condyle width and height and ICN width indices were significantly greater in the low risk compared to the high risk breeds (P<0.01 for all parameters). In contrast, the pro (P=0.003) and active (P=0.007) forms of MMP-2 and sulphated GAGs (P=0.0002) were significantly greater in the impinged areas of the ACLs of the rupture predisposed breeds. CONCLUSIONS: Impingement by the ICN on the ACLs of the high risk breeds may result in increased collagen remodelling and increased sulphated GAG deposition, causing reduced structural integrity of the ligament. Altered ACL composition may predispose the ligament to increased laxity leading to joint degeneration and OA. This may have a comparative implication for pathogenesis of ACL rupture in humans. PMID- 16242972 TI - Early post-traumatic osteoarthritis-like changes in human articular cartilage following rupture of the anterior cruciate ligament. AB - OBJECTIVE: Injury to the anterior cruciate ligament (ACL) frequently leads to post-traumatic osteoarthritis (OA). In this study we determined whether early degenerative changes characteristic of idiopathic OA are induced in articular cartilage following ACL injury. METHODS: A small sample of femoral articular cartilage was removed at surgery, as part of ACL reconstruction, from a total of 50 patients with ACL injuries. Of these, 28 underwent surgery less than 1 year post-injury. Control cartilages were obtained from the same site from 21 persons at autopsy. All cartilages were examined for molecular changes. The content of type II collagen, its cleavage by collagenases and its denaturation were determined by immunoassay. The total content of glycosaminoglycan (GAG), which is principally aggrecan, was measured colorimetrically. Data were expressed per unit DNA (GAG and collagen content) or as a percentage of total collagen cleaved or denatured. Other cartilages from the same site (8 controls, 12 less than 1 year and 8 more than 1 year post-injury) were frozen sectioned and examined histologically to determine by Mankin grading cartilage degeneration. RESULTS: Histological analyses revealed that control subjects exhibited staining for proteoglycan, which was reduced in some patients following ACL rupture. Degeneration of the articular surface was sometimes observed 1 year after ACL rupture. Although the Mankin grade increased with time after rupture these changes were not significant. Immunoassays, however, revealed an increase in GAG content within 1 year which was maintained after 1 year although no longer significant. No changes in total type II collagen content were observed during the period of study. However, there were significant increases in the denaturation and cleavage of type II collagen less than and more than 1 year post ACL rupture. Total type II collagen content was directly correlated with GAG content in all three groups, with the significance being weakest at more than 1 year. After 1 year an inverse correlation was observed between total type II collagen content and collagen cleavage as well as denaturation. CONCLUSIONS: These observations reveal that joint instability resulting from ACL injury rapidly results in degenerative changes characteristic of those seen in idiopathic OA at arthroplasty and in experimental OA following ACL surgery. These changes may contribute to the development of post-traumatic OA that is commonly observed following ACL injury. The observations support and extend conclusions from other studies on human and animal articular cartilage and synovial fluids post-ACL injury that have revealed a rapid onset of damage to type II collagen and an initial increase in proteoglycan content characteristic of experimental OA post-ACL injury. This study provides direct evidence for the rapid development of degenerative changes characteristic of OA following ACL injury. PMID- 16242973 TI - Heterogeneity in patellofemoral cartilage adaptation to anterior cruciate ligament transection; chondrocyte shape and deformation with compression. AB - OBJECTIVE: The purpose of this study was to determine if the opposing cartilages of the feline patellofemoral joint adapted differently to short-term anterior cruciate ligament transection (ACL-T) and if the magnitude of chondrocyte deformation upon tissue loading was altered under ACL-T conditions compared to contralateral controls. In situ static compression of physiological magnitude was applied to the feline patellofemoral cartilage 16 weeks post-ACL-T and cartilage and chondrocyte deformation were evaluated by histomorphometry. DESIGN: Six adult cats were euthanized 16 weeks after unilateral ACL-T. A peak surface pressure of 9 MPa was applied to the fully intact patella and femoral groove cartilages. After in situ fixation under compression, sections from the centre of the indent and from an adjacent unloaded area of the cartilages were analysed. Chondrocyte shape, size, clustering and volumetric fraction were quantified. RESULTS: Experimental patellar articular cartilage was thicker, contained larger chondrocytes that were more frequently arranged in clusters and had, on average, a larger chondrocyte volumetric fraction compared to contralateral controls. In contrast, the experimental femoral groove cartilage demonstrated little adaptation to ACL-T. CONCLUSIONS: The patellar articular cartilage adapts to short-term ACL-T to a greater extent than femoral groove cartilage. We speculate that differences in the histological parameters of control tissues, such as cartilage thickness and the magnitude and depth distribution of chondrocyte shape, size and volumetric fraction may contribute to predisposing patellar cartilage, and not femoral groove cartilage, to adaptation after ACL-T. PMID- 16242974 TI - Induction of MMP-13 expression by soluble human glucocorticoid-induced tumor necrosis factor receptor in fibroblast-like synovial cells. AB - OBJECTIVE: We tested the hypothesis that human glucocorticoid-induced tumor necrosis factor receptor (hGITR/TR11) expressed on the surface of activated CD4(+) T cells is responsible for up-regulating the production of matrix metalloproteinase (MMP)-13 by fibroblast-like synoviocytes (FLSs). METHODS: The level of MMP-13 was measured by Western blot and reverse transcriptase polymerase chain reaction (RT-PCR). Expressions of hGITR ligand (hGITRL) on the surface of FLSs and hGITR on the surface of human CD4(+) T cells were analyzed by flow cytometry and RT-PCR. Neutralizing antibodies (Abs) were used to block hGITRL and hGITR on the surface of FLSs and human CD4(+) T cells, respectively. Human CD4(+) T cells were cocultured with FLSs to facilitate interaction between hGITR on CD4(+) T cells and hGITRL on FLSs. RESULTS: Soluble hGITR (shGITR) stimulated FLSs to produce MMP-13, and blockade of hGITRL reduced this effect. Direct contact between activated CD4(+) T and FLSs also induced the production of MMP 13, and neutralization of hGITR on activated CD4(+) T cells during coculture decreased the amount of MMP-13 produced by FLSs. CONCLUSION: shGITR stimulated FLSs to produce MMP-13 via a signal through hGITRL. Direct contact between activated CD4(+) T cells and FLSs facilitated hGITR-hGITRL interaction, and resulted in inducing MMP-13. This effect may increase tissue destruction in chronic inflammation such as rheumatoid arthritis (RA). PMID- 16242975 TI - Invasive growth and topoisomerase-switch induced by tumorous extracellular matrix in osteosarcoma cell culture. AB - Osteosarcoma cells are capable of extracellular matrix (ECM) synthesis. The ability of ECM to trigger the proliferation of a novel osteosarcoma cell line (OSCORT) was tested in this study in relation to a known tumor ECM, isolated from Engelbreth-Holm-Swarm (EHS) sarcoma (EHS-ECM). OSCORT was grown in monolayer, in EHS-ECM and in ECM deposited by the cells (OSCORT-ECM). Both EHS-ECM and OSCORT ECM increased the proliferation and migration of OSCORT cells. Among the ECM biopolymers, heparan sulfate proteoglycan (HSPG) and fibronectin enhanced invasive growth, collagen type IV reduced it, while laminin had no effect. Among the ECM components HSPG and collagen IV increased both the synthesis and activation of collagenase type IV, and all the ECM components substantially increased beta1 integrin levels in the cells. The majority of ECM biopolymers decreased the level of topoisomerase I (except laminin) and elevated topoisomerase II (except fibronectin) in OSCORT. The switch in the ratio between the activities of topoisomerases I and II was mainly due to HSPG. The HSPG synthesized by OSCORT cells is described as agrin, which is a novel finding. The present study showed that HSPG (agrin) showed the most remarkable stimulatory action on the growth and migration of OSCORT cells. HSPG-induced topoisomerase II induction deserves further experimentation, to discover its relevance to tumor progression. PMID- 16242977 TI - Rolipram reverses scopolamine-induced and time-dependent memory deficits in object recognition by different mechanisms of action. AB - In this study, the effect of the selective phosphodiesterase type 4 (PDE4) inhibitor rolipram on memory performance was investigated using the object recognition task. First, three doses of rolipram (0.01, 0.03 or 0.1 mg/kg) were tested with a 24h delay between the learning (T1) and the test (T2) trial. Doses of rolipram were injected at different time points (30 min before T1, immediately after T1 or 3 h after T1). In a second experiment, the effects of rolipram (0.03, 0.1 or 0.3 mg/kg) were tested in combination with scopolamine (0.1 mg/kg) applying a 1 h delay between trials. Both substances were administered 30 min before T1. Using a 24h interval, rolipram showed an improvement in long-term memory performance when injected 3 h after T1 at a dose of 0.03 mg/kg. Further, rolipram reversed the scopolamine-induced short-term memory deficit at a dose of 0.1 mg/kg. Although the improved memory performance in both conditions is likely to be explained by elevated cAMP levels, two separate working mechanisms might explain these effects. PMID- 16242976 TI - In vivo expression and characteristics of novel alpha-D-mannose-rich glycoprotein markers of apoptotic cells. AB - We recently established that an increased expression of alpha-D-mannose (Man)- and beta-D-galactose-rich plasma membrane glycoproteins (GPs) is characteristic for apoptotic cells in vitro [Bilyy, R.O., Stoika, R.S., 2003. Lectinocytochemical detection of apoptotic murine leukemia L1210 cells. Cytometry 56A, 89-95]. It was independent of cell line or apoptosis-inducing agent, and can therefore be considered as a selective marker for identification and isolation of apoptotic cells [Bilyy, R.O., Antonyuk, V.O., Stoika, R.S., 2004. Cytochemical study of role of alpha-D-mannose- and beta-D-galactose-containing glycoproteins in apoptosis. J. Mol. Histol. 35, 829-838]. The main goals of the present study were: (1) to determine whether an increased expression of specific GPs also takes place after apoptosis induction in vivo; and (2) to identify additional characteristics of the membrane GP markers of the apoptotic cells. To reach these goals, we studied the expression of alpha-Man-rich membrane GPs in murine leukemia L1210 cells inoculated into abdominal cavities of mice which were then subjected to the action of apoptosis inducer doxorubicin. Another experimental model used in the present work was splenocytes obtained from mice treated with dexamethasone. Lectin-affinity chromatography and PAGE electrophoresis, or PAGE electrophoresis and lectinoblot analysis were applied for isolation of plasma membrane GPs (34 kDa, and high M(W) of approximately 600 and 800 kDa) whose expressions were increased during apoptosis. Triton X-114 treatment of cell membrane samples showed that the apoptotic cell-specific GPs were localized in the peripheral and integral compartments of plasma membrane. Apoptosis in vitro and in vivo was accompanied by an increased expression of the same GP, identified by MALDI-TOF MS analysis as the microtubule-actin cross-linking factor 1. Other GPs, whose expressions were also increased at apoptosis, were similarly identified as G-protein beta-subunit like (Acc# BAA06185.1) and dystonin isoform beta. PMID- 16242978 TI - Haemodynamic study examining the response of venous blood flow to electrical stimulation of the gastrocnemius muscle in patients with chronic venous disease. AB - OBJECTIVES: The aim of this study was to explore the option of stimulating calf muscle contraction through externally applied neuromuscular electrical stimulation (NMES) and to measure venous blood flow response to this stimulation. METHODS: Ten patients with class 6 chronic venous disease (CEAP clinical classification) were recruited. Measurements of peak venous velocities in the popliteal vein were recorded by Duplex scanning in response to six test conditions; 1. Standing, 2. Voluntary calf muscle contraction, 3. Standing with NMES applied, 4. Standing with compression bandaging applied to the leg, 5. Voluntary calf muscle contraction with compression bandaging applied to the leg, 6. Stationary with compression bandaging applied to the leg and NMES applied. Comfort assessment was completed using visual analogue scales at each test stage and on study completion each patient completed a short structured interview to determine comfort and acceptability of NMES. Statistical analyses were carried out using SPSS, Version 9. Non-parametric testing was used in all analyses using the Wilcoxon Signed Ranks Test for paired samples. RESULTS: There was a significant increase in venous velocities on voluntary contraction of the calf muscle (median resting vel 7.3 cm/s; voluntary contraction median 70 cm/s) and with the introduction of NMES, both with compression (median velocity 15 cm/s, p = 0.005 Wilcoxon) and without compression (median velocity 13 cm/s, p = 0.005 Wilcoxon). The greatest increase with NMES was when combined with compression bandaging. All patients reported the stimulus as an acceptable treatment option with 90% reporting NMES as comfortable. CONCLUSIONS: Healing rates in venous ulceration with the application of compression bandaging remain between 50 and 70%. This study shows a positive haemodynamic response to NMES. Further research is needed to quantitatively measure the effect of NMES on ulcer healing. PMID- 16242979 TI - Changes in internal carotid blood flow after CEA evaluated by transit-time flowmeter. AB - AIM: The aim of this study was to investigate whether there was an association between the degree of the stenosis of the internal carotid artery (ICA) and post operative increase of blood flow. METHODS AND MATERIALS: In 200 out of 660 patients undergoing carotid endarteriectomy (CEA) for a high-degree ICA stenosis, pre-operatively a bilateral selective carotid and intracerebral angiography was performed. The degree of the ipsilateral and contralateral stenosis was digitally assessed by using computer software according to the CC-Index. Intraoperatively, the pressure ratio over the stenosis (ICA/CCA) was measured by direct arterial puncture. Blood flow in the ICA was measured before and after CEA with an ultrasound flowmeter using the transit-time principle. These findings were correlated to the degree of stenosis revealed by angiographic analysis and the pressure ratio. RESULTS: Before CEA the median blood flow in the ICA was 171 ml/min (range 620 ml/min) with a significant (p<0.001) post-operative increase to 250 ml/min (range 875 ml/min). The median relative increase of flow (post-flow pre-flow/pre-flow) was 42%. The pre-CEA flow volumes were dependent on the degree of stenosis and also the pressure ratio. The increase of flow following CEA correlated better with pressure ratio (r=-0.435, p<0.001), than the stenosis severity (r=0.319, p<0.001). Analysis of variance identified only the pressure gradient as an independent determinant of flow changes following CEA. CONCLUSIONS: The blood flow increase following CEA is mainly determined by the pressure gradient across the stenosis. PMID- 16242980 TI - Antioxidant nutrient intake and supplements as potential moderators of cognitive decline and cardiovascular disease in obstructive sleep apnea. AB - Cognitive deficits and cardiovascular disease (CVD) are comorbid conditions frequently associated with obstructive sleep apnea (OSA). Oxygen free radical release and its differential regulation of cytokine synthesis and immune modulation resulting from OSA-related hypoxic events have been hypothesized as the underlying mechanism(s) for the cognitive deficits and CVD in OSA. A number of studies have suggested that increased levels of oxidative stress and/or antioxidant deficiencies may also be risk factors in cognitive decline and CVD. The influence of antioxidant nutrients and supplements, such as Vitamins B6, B12, C, E, folic acid, alpha-lipoic acid and Coenzyme Q(10) on cognitive decline and CVD have been investigated. The influence of antioxidant nutrients or supplements on OSA remains to be investigated. Even if dietary or supplemental antioxidants do not prove to be effective therapies for OSA, dietary assessment and prescription to increase dietary intake of neuro- and cardio-protective nutrients may make it possible to reduce some of the cognitive and cardiovascular sequelae associated with OSA. PMID- 16242981 TI - Phosphodiesterase IV and neutral endopeptidase in airways from developing and allergen sensitized rabbits. AB - Phosphodiesterase IV (PDE IV) and neutral endopeptidase (NEP) may modulate the neurally mediated nonadrenergic noncholinergic inhibitory (NANCi) response. This response is not present in normal rabbits until 2 weeks of age. Allergen sensitization and challenge of fully grown 13-week old rabbits decreases the NANCi response. Our goal was to assess NANCi as a function of age and allergen sensitization. Tracheal smooth muscle (TSM) rings from normal 1-, 2-, and 13-week old rabbits plus ragweed immune as well as ragweed immune/challenged (I/C) 13 week old rabbits were assessed. Colorimetric assays of PDE IV and NEP activity were conducted on TSM from each group. NANCi responses were obtained in the presence and absence of Ro 20-1724 (PDE IV inhibitor) and/or phosphoramidon (Phos; NEP inhibitor) after contraction of TSM with neurokinin A. In normal TSM, there was no difference in PDE IV as a function of age. Conversely, NEP decreased significantly from 1 to 13 weeks of age. The immune and I/C groups had decreases in NEP and increases in PDE IV that were significant. Neither Ro 20-1724 nor Phos alone or together increased NANCi responses in TSM from 1- or 2-week old rabbits. However, both enhanced relaxation in TSM from normal, immune, and I/C 13-week old rabbits with an additive effect when drugs were combined. This work demonstrates (1) normal maturational changes in NEP but not PDE IV within TSM of this species; (2) modulation of the NANCi response by inhibitors of PDE IV and NEP in 13- but not 2-week old rabbits; (3) increased PDE IV and decreased NEP levels in the immune and I/C groups with reconstitution of NANCi responses by the combination of inhibitors. We conclude that mediation of the NANCi response is different in normal 2- and 13-week old rabbits. Both PDE IV and NEP modulated relaxation in fully grown rabbits, but had no effect at the younger age. Furthermore, both ragweed sensitization alone and ragweed challenge of immune rabbits altered NANCi via increases in PDE IV and decreases in NEP. PMID- 16242982 TI - Application of sequential integration for metabolic engineering of 1,2 propanediol production in yeast. AB - The yeast species Saccharomyces cerevisiae was engineered to produce 1,2 propanediol (1,2-PD) using the delta/UB sequential gene integration method. To study the effects of increased copy number, 2 genes (mgs and gldA from Escherichia coli) were sequentially integrated into the chromosomes of S. cerevisiae strains of opposite mating type. The resulting strains (containing 0-3 copies of either mgs or gldA) were mated to create all possible combinations of the 2 genes introduced for 1,2-PD production. Enzyme activities were generally correlated with copy number, although there was greater variation in GldA activity in the diploid cells. The integrated genes were confirmed by Southern blot and 1,2-PD production was analyzed by HPLC. The strain containing 3 copies of mgs and gldA showed the highest level of 1,2-PD; however, 1,2-PD concentration was not clearly related to gene copy number. 1,2-PD production did correlate with Mgs specific activity, and high GldA specific activity was found to be inhibitory. PMID- 16242983 TI - Signal transduction and metabolic flux of beta-thujaplicin and monoterpene biosynthesis in elicited Cupressus lusitanica cell cultures. AB - beta-Thujaplicin is an antimicrobial tropolone derived from geranyl pyrophosphate(GPP) and monoterpene intermediate. Yeast elicitor-treated Cupressus lusitanica cell cultures accumulate high levels of beta-thujaplicin at early stages and other monoterpenes at later stages post-elicitation. The different regulation of beta-thujaplicin and monoterpene biosynthesis and signal transduction directing metabolic flux to beta-thujaplicin firstly and then shifting metabolic flow from beta-thujaplicin to other monoterpene biosynthesis were investigated. The earlier rapid induction of beta-thujaplicin accumulation and a later stimulation of monoterpene biosynthesis by yeast elicitor are in well agreement with elicitor-induced changes in activity of three monoterpene biosynthetic enzymes including isopentenyl pyrophosphate isomerase, GPP synthase, and monoterpene synthase. Yeast elicitor induces an earlier and stronger beta thujaplicin production and monoterpene biosynthetic enzyme activity than methyl jasmonate (MeJA) does. Profiling all monoterpenes produced by C. lusitanica cell cultures under different conditions reveals that beta-thujaplicin biosynthesis parallels with other monoterpenes and competes for common precursor pools. Yet beta-thujaplicin is produced pre-dominantly at early stage of elicitation whereas other monoterpenes are mainly accumulated at late stage while beta-thujaplicin is metabolized. It is suggested that yeast elicitor-treated C. lusitanica cells preferentially accumulate beta-thujaplicin as a primary defense and other monoterpenes as a secondary defense. Inhibitor treatments suggest that immediate production of beta-thujaplicin post-elicitation largely depends on pre-existing enzymes and translation of pre-existing transcripts as well as recruitment of precursor pools from both the cytosol and plastids. The later beta-thujaplicin and other monoterpene accumulation strictly depends on active transcription and translation. Induction of beta-thujaplicin production and activation of monoterpene biosynthetic enzymes by elicitor involves similar signaling pathways, which may activate early beta-thujaplicin production and later monoterpene biosynthesis and induce a metabolic flux shift from beta-thujaplicin to monoterpene accumulation. PMID- 16242984 TI - Long-term treatment of patients with mild and classical phenylketonuria by tetrahydrobiopterin. AB - Tetrahydrobiopterin (BH4), the natural cofactor of phenylalanine hydroxylase (EC 1.14.16.1), can reduce blood phenylalanine (Phe) in BH4 sensitive patients with hyperphenylalaninemia (McKuisick 261600). We report on the long-term treatment of eight patients with mild and classical phenylketonuria (blood Phe levels maximum blood Phe levels between 771 and 1500 micromol/L) using BH4 at a dosage of 8-12 mg/kg BW per day. In all patients reduction of blood Phe was >30% after BH4 loading test. Three patients were treated from birth by BH4 only, five after initial low Phe dietary treatment. Seven of them continue to be on BH4 treatment only, one has a relaxed low protein diet. No side effects could be observed (longest observation time 5 years), somatic and psychomotor development were normal. The main problem of BH4 treatment is finding an optimal dosage at different ages and an under special conditions like infectious diseases. There is evidence that in some patients BH4 treatment may allow a more relaxed low protein diet showing positive effects on weight gain and quality of life. Further controlled studies are necessary not only to rule out any side effects but also for optimizing treatment strategies with BH4 treatment in mild phenylketonuria. PMID- 16242985 TI - Antibody response achieved by different rabies prophylaxis methods. PMID- 16242986 TI - Cytotoxic effect of Shiga toxin-2 holotoxin and its B subunit on human renal tubular epithelial cells. AB - Shiga toxin-producing Escherichia coli produces watery diarrhea, hemorrhagic colitis and hemolytic-uremic syndrome (HUS). In Argentina, HUS is the most common cause of acute renal failure in children. The purpose of the present study was to examine the cytotoxicity of Stx type 2 (Stx2 holotoxin) and its B subunit (Stx2 B subunit) on human renal tubular epithelial cells (HRTEC), in the presence and absence of inflammatory factors. Cell morphology, cell viability, protein synthesis and apoptosis were measured. HRTEC are sensitive to both Stx2 holotoxin and Stx2 B subunit in a dose- and time-dependent manner. IL-1, LPS and butyrate but not TNF, IL-6 and IL-8, increased the Stx mediated cytotoxicity. The effects of Stx2 B subunit appear at doses higher than those used for Stx2 holotoxin. Although the physiological importance of these effects is not clear, it is important to be aware of any potentially toxic activity in the B subunit, given that it has been proposed for use in a vaccine. PMID- 16242987 TI - A reliable technique for avoiding the median nerve during carpal tunnel injections. AB - Carpal tunnel injections are widely performed for diagnostic and therapeutic purposes. Injury to the median nerve is a serious and fairly common complication. There is no consensus regarding the safest injection site. The objective of this study was to determine the safest injection site based on anatomical data. During 124 endoscopic procedures for median nerve release at the carpal tunnel, we measured the distances separating the median nerve, palmaris longus (PL), flexor carpi radialis (FCR), and flexor carpi ulnaris 1 cm proximal to the wrist crease. The edge of the median nerve extended medially beyond the PL tendon in 82 (88%) hands. Thus, needle insertion within 1 cm of either edge of the PL tendon may cause median nerve injury; with injection sites located further toward the medial edge, the ulnar pedicle may be at risk. Consequently, we recommend that carpal tunnel injections be performed through the FCR tendon. PMID- 16242988 TI - Bacterial histidine kinase as signal sensor and transducer. AB - Adaptation to an environmental stress is essential for cell survival in all organisms, from E. coli to human. To respond to changes in their surroundings, bacteria utilize two-component systems (TCSs), also known as histidyl-aspartyl phosphorelay (HAP) systems that consist of a histidine kinase (HK) sensor and a cognate response regulator (RR). While mammals developed complex signaling systems involving serine/threonine/tyrosine kinases in stress response mechanisms, bacterial TCS/HAP systems represent a simple but elegant prototype of signal transduction machineries. HKs are known as a seductive target for anti bacterial therapeutic development, because of their significance in pathological virulence in some bacteria such as Salmonella enterica. Recent molecular and structural studies have shed light on the molecular basis of the signaling mechanism of HK sensor kinases. This review will focus on recent advancements in structural investigation of signal sensing and transducing mechanisms by HKs, which is critical to our understanding of bacterial biology and pathology. PMID- 16242989 TI - snRNAs as the catalysts of pre-mRNA splicing. AB - The spliceosome, the gigantic molecular machine that performs pre-mRNA splicing in eukaryotes, contains over 200 different proteins and five RNA molecules. The central role played by the spliceosomal RNAs in splicing has led to the hypothesis that, like the ribosome, the spliceosome is an RNA-centric enzyme and a relic from the RNA world. Recent structural studies have provided the first glimpses of the structural features of spliceosomal RNAs, and mutational analyses in vivo and in vitro have uncovered new functional roles for a catalytically essential domain. An emerging model for the active site of group II introns, a closely related class of natural ribozymes, is likely to provide a wealth of insights on structure and function of the active site of the spliceosome. PMID- 16242990 TI - Nucleotide analogues to investigate RNA structure and function. AB - RNA plays an essential cellular role in nearly every aspect of the transmission and expression of genetic information, including regulatory roles that have significance for cellular development. Access to RNA bearing synthetic modifications has allowed biological chemists to probe deep into the inner workings of cellular processes. Here, we describe recent advances in harnessing the power of nucleotide analogues to obtain mechanistic and biological insights into RNA structure, function and dynamics. PMID- 16242991 TI - Archaeal DNA replication and repair. AB - Since the first archaeal genome was sequenced, much attention has been focused on the study of these unique microorganisms. We have learnt that although archaeal DNA metabolic processes (replication, recombination and repair) are more similar to the metabolic processes of Eukarya than those of Bacteria, Archaea are not simply 'mini Eukarya'. They are, in fact, a mosaic of the eukaryal and bacterial systems that also possess archaeal-specific features. Recent biochemical and structural studies of the proteins that participate in archaeal DNA replication and repair have increased our understanding of these processes. PMID- 16242992 TI - Archaebacteria (Archaea) and the origin of the eukaryotic nucleus. AB - The eukaryotic nucleus is a unique structure. Because it lacks an obvious homologue or precursor among prokaryotes, ideas about its evolutionary origin are diverse. Current attempts to derive the nuclear membrane focus on invaginations of the plasma membrane in a prokaryote, endosymbiosis of an archaebacterium within a eubacterial host, or the origin of a genuinely new membrane system following the origin of mitochondria in an archaebacterial host. Recent reports point to ways in which different ideas regarding the origin of the nucleus might someday be discriminated. PMID- 16242993 TI - Methyl-coenzyme M reductase and the anaerobic oxidation of methane in methanotrophic Archaea. AB - Recent biochemical and metagenomic data indicate that not yet cultured Archaea that are closely related to methanogenic Archaea of the order of Methanosarcinales are involved in the anaerobic oxidation of methane in marine sediments. The DNA from the methanotrophic Archaea has been shown to harbor gene homologues for methyl-coenzyme M reductase, which in methanogenic Archaea catalyses the methane-forming reaction. In microbial mats catalyzing anaerobic oxidation of methane, this nickel enzyme has been shown to be present in concentrations of up to 10% of the total extracted proteins. PMID- 16242994 TI - Further evaluation of the skin micronucleus test: results obtained using 10 polycyclic aromatic hydrocarbons. AB - The standard in vivo micronucleus (MN) test detects clastogenicity in hematopoietic cells and is not suitable for detecting chemicals that target the skin. Previously, we have developed an in vivo rodent skin MN test that is simple to perform and can be applied to several laboratory animals, including the hairless mouse-a species whose use simplifies the procedure of skin testing. In this paper, we report new data that confirms the predictive ability of the test. Following the application of 10 polycyclic aromatic hydrocarbons (7,12 dimethylbenz[a]anthracene; 3-methylcholanthrene; benzo[a]pyrene; dibenz[a,h]anthracene; benz[a]anthracene; dibenz[a,c]anthracene; chrysene; benzo[e]pyrene; pyrene; anthracene) with various degrees of genotoxicity to the dorsal skin of hairless mice, we found that these compounds caused MN production that in general correlated with their reported carcinogenicity. We believe that this test will be useful in detecting skin clastogens that test negative when analyzed using the standard micronucleus test. PMID- 16242995 TI - Epidemic spread of recombinant noroviruses with four capsid types in Hungary. AB - BACKGROUND: Noroviruses are common pathogens in gastro-enteritis outbreaks in humans worldwide. Noroviruses are genetically diverse group of viruses with multiple genogroups (GG) and genotypes. More recently, naturally occurring recombinant noroviruses were described. These viruses had a distinct polymerase gene sequence (designated GGIIb/Hilversum) and were disseminated through waterborne and food-borne transmission in Europe. OBJECTIVES: Our aim was to characterize these emerging recombinant noroviruses causing outbreaks of gastro enteritis in Hungary. STUDY DESIGN: From January 2001 to May 2004, samples containing "GGIIb/Hilversum polymerase" (GGIIb-pol) were selected for analysis of the viral capsid region (ORF2) by reverse transcription-polymerase chain reaction (RT-PCR) and sequencing. RESULTS: Thirty-four (14.4%) of 236 confirmed norovirus outbreaks were caused by the variant lineage with the GGIIb-pol. Four different recombinants were detected with capsids of Hu/NLV/GGII/Mexico/1989 (n=9, 43%), Hu/NLV/GGII/Snow Mountain/1976 (n=6, 28%), Hu/NLV/GGII/Hawaii/1971 (n=4, 19%) and Hu/NLV/GGII/Lordsdale/1993 (n=1, 5%). CONCLUSIONS: In Hungary, emerging recombinant noroviruses became the second most common norovirus variants-next to GGII-4/Lordsdale virus-causing epidemics of gastroenteritis in the last 4 years. PMID- 16242996 TI - Proceedings of the International Symposium in Emerging Trends in Tuberculosis Research, November 15-17, 2004, New Delhi, India. PMID- 16242997 TI - Mechanistic studies of sequential injection of cationic liposome and plasmid DNA. AB - We previously reported that sequential injection of cationic liposome and plasmid DNA leads to notably reduced inflammatory toxicity and improved transfection in the lung (Y. Tan et al., 2001, Mol. Ther. 3, 673-682). The purpose of the current study was to explore the mechanism involved in sequential injection. It was observed that sequential injection resulted in dramatically lower DNA uptake by the liver and higher DNA levels in the lung than the lipoplex injection. In vitro experiments with macrophage cells further showed that sequential addition of liposomes and DNA could diminish the cellular uptake of DNA by these cells. The contributions of serum to the enhanced bioactivity and decreased toxicity were examined by injecting mice with samples of premixed liposome with serum and then DNA (LSD sample), and the resulting activities were compared to those obtained with injection of lipoplex-serum mixtures (LDS sample). LSD yielded 80% lower TNF alpha levels and over 10-fold higher transfection than lipoplex, which is consistent with the reported findings with sequential injection. In contrast, LDS resulted in the same TNF-alpha levels and comparable transfection with lipoplex. Thus, the results suggest that the primary interaction of serum with liposome is a critical factor contributing to the superior activity and reduced toxicity of sequential injection. Studies on the interaction between mouse serum, liposomes, and DNA showed that DNA could bind negatively charged liposome-serum complex to form a ternary complex, which has a density similar to that of the ternary complex formed between lipoplex with serum. Further in vitro tests showed that LSD and LDS were similar in particle size and protein content, but different in protein composition as observed by 2-D gel electrophoresis. In addition, DNA in LSD was more readily displaced by dextran sulfate, an anionic polymer, than in LDS. The above findings suggest that the inhibition of opsonin protein binding on the particle surface with the sequential injection may contribute to the reduced macrophage uptake and cytokine induction and that the high ability of DNA release from the particles formed after sequential injection may contribute to the improved lung gene transfection. PMID- 16242998 TI - Activation of refractory T cell responses against hepatitis C virus core protein by ablation of interfering hydrophobic domains. AB - Hepatitis C virus (HCV) is the major pathogen of chronic hepatitis and liver disease, but currently there are no prophylactic HCV vaccines available. The HCV core protein-encoding sequence is among the most conserved genes in the HCV genome, making it a prime candidate for a component of a vaccine. The core protein localizes to the endoplasmic reticulum (ER) through a C-terminal hydrophobic region that is cotranslationally inserted into the ER membrane. Here we show that removal of the C-terminal hydrophobic region confers nuclear localization and enhances proteasomal degradation of the core protein in mammalian cells. This efficient protein proteolysis induces enhanced core specific CD8(+) T cell responses in BALB/c mice immunized with plasmids expressing C-terminal deletions of the HCV core protein. These results suggest that more potent HCV vaccines can be achieved by targeting the core protein for proteasomal degradation by deletion of its C-terminal hydrophobic domain. PMID- 16242999 TI - Generation, culture, and differentiation of human embryonic stem cells for therapeutic applications. AB - Embryonic stem (ES) cells, derived from the inner cell mass of the mammalian blastocyst, can continuously proliferate in an undifferentiated state and can also be induced to differentiate into a desired cell lineage. These abilities make ES cells an appealing source for cell replacement therapies, the study of developmental biology, and drug/toxin screening studies. As compared to mouse ES cells, human ES cells have only recently been derived and studied. Although there are many differences in properties between mouse and human ES cells, the study of mouse ES cells has provided important insights into human ES cell research. In this review, we describe the advantages and disadvantages of methods used for human ES cell derivation, the expansion of human ES cells, and the current status of human ES cell differentiation research. In addition, we discuss the endeavor that scientists have undertaken toward the therapeutic application of these cells, which includes therapeutic cloning and the improvement of human ES cell culture conditions. PMID- 16243000 TI - Sexuality in women with epilepsy. AB - Most women with epilepsy maintain normal reproductive cycles and sexual lives. However, a significant minority, approximately 20-30%, have some degree of sexual dysfunction, including problems with seizure exacerbation, libido, arousal, and orgasm. Fluctuating hormone levels may contribute to an array of reproductive cycling abnormalities. With regard to sexual dysfunction, there is some evidence of reduced genital blood flow in women with temporal lobe epilepsy. Other studies suggest that psychosocial factors, such as depression, feeling stigmatized, and being anxious about having seizures during sex, may contribute to the higher rates of sexual dysfunction in this patient population. Some antiepileptic drugs may adversely affect normal reproductive cycling and sexual function, particularly drugs that increase serotonergic transmission. Conversely, resective epilepsy surgery has been shown to restore sexual function. Treatments for sexual dysfunction include testosterone replacement, although transdermal testosterone replacement is not yet approved by the Food and Drug Administration for women. Given the possibility that women with epilepsy may experience inadequate vasocongestion during arousal, sildenafil may have a useful role, though it has not proved effective for women in general. This review focuses on potential sexual problems that are faced by women with epilepsy, with the suggestion that proper treatment may alleviate these problems. PMID- 16243001 TI - Human sexuality, sex hormones, and epilepsy. AB - The function of the hypothalamic-pituitary axis (HPA), including the production of luteinizing hormone, follicle-stimulating hormone, gonadotropin-releasing hormone, and prolactin, and the concentrations and metabolism of its end products, such as estrogen, testosterone, and dehydroepiandrosterone, appear to be modified in many people with epilepsy. Effects of the disorder itself and effects of antiepileptic drugs (AEDs) both appear to contribute to these hormonal alterations, which may be associated with sexual dysfunction. Focal epileptic discharges from the temporal lobe may affect HPA function, as is suggested by the normalization of androgen levels seen in men with temporal lobe epilepsy who become seizure-free after surgery. Hepatic enzyme-inducing AEDs such as carbamazepine and phenytoin may be most clearly linked to altered metabolism of sex steroid hormones, but valproic acid, an enzyme inhibitor, has also been implicated in the causation of reproductive endocrine abnormalities. Polycystic ovaries and polycystic ovarian syndrome (PCOS) are widely believed to be common in women with epilepsy, but the actual prevalence and the pathogenesis of PCOS in this population are disputed. Hormonal changes and sexual dysfunction need to be addressed in any comprehensive approach to epilepsy management, as well as any comprehensive epilepsy research program. Avoidance of enzyme-inducing AEDs and achievement of freedom from seizures as the goal of treatment are strongly recommended. PMID- 16243002 TI - Neuroprotection in epilepsy: the Holy Grail of antiepileptogenic therapy. PMID- 16243003 TI - Assessment of sexual functioning: sexual history taking for health care practitioners. AB - It is important for clinicians to inquire about sexual functioning as part of routine mental and physical health care. General guidelines for taking a skillful and sensitive sexual history are given, including ways to initiate discussions and make the patient feel more at ease. In addition, a sample comprehensive sexual history is presented. This is a revised version of the Integrated Model of Sexual Therapy developed by Helen Singer Kaplan, M.D., Ph.D. It includes a large number of sample questions, which are organized according to such categories as exploring the chief complaint, the sexual status examination, and the medical, psychiatric, family, psychosexual, and relationship histories. Additional questions pertaining to epilepsy and the prevention of sexually transmitted diseases are also included. PMID- 16243004 TI - Reproductive and sexual dysfunction in men with epilepsy. AB - Disturbances of reproductive and sexual health are common in people with epilepsy. Their etiology is not well understood but appears to be multifactorial, and both epilepsy itself and drugs used to treat it are implicated. Physiologically, sex steroid hormone levels, the hypothalamic-pituitary axis, and testicular function can be affected in men with epilepsy. Psychosocial complications associated with epilepsy can also affect reproductive health and sexuality. Clinicians need to investigate such problems carefully, both because of their multifactorial nature and because patients and physicians alike may often fail to recognize or be reluctant to acknowledge them; in particular, patients whose epilepsy had its onset before puberty may lack subjective awareness of impairments of sexual response and function. Treatments for reproductive and sexual dysfunction in men with epilepsy have been inadequately studied. Modalities such as medications for erectile dysfunction and surgery may be useful. Therapy with exogenous testosterone and an aromatase inhibitor may be helpful for men with epilepsy and sexual dysfunction due to testosterone deficiency. PMID- 16243005 TI - Conceptualizing the world: lessons from history. AB - Throughout history, philosophers, scientists, and other scholars have named and organized the salient elements of the world. These efforts have led to conceptualizations that differ widely in both content and form. This paper argues that all conceptualizations are biased, both because they depend on the purposes for which they have been created, and because they are closely tied to the world view of their designers. This bias needs to be recognized, and its consequences need to be addressed if the conceptualizations are to be used for purposes other than those for which they were designed. The paper begins with a brief overview of the disciplines that have been concerned with conceptualizing particular domains. This is followed by a non-exhaustive, illustrative, historical perspective, and the paper concludes with the interesting case study of biological taxonomy. PMID- 16243006 TI - Tree disagreement: measuring and testing incongruence in phylogenies. AB - The branching patterns of phylogenetic trees often disagree even when they have been constructed using different portions of the same data. This phylogenetic discord (incongruence) can be explained by real differences in evolutionary process or history, but also may be due simply to random chance or sampling error. Techniques for measuring and testing the significance of phylogenetic incongruence are used widely in systematic biology, and are necessary when considering genome-scale datasets composed of multiple genes that may or may not have different histories. They are also applicable wherever tree algorithms are used for ordering and interpreting data (e.g., DNA microarrays). Here, I review the different incongruence tests and use them to test the phylogenetic discord of a potentially mobile genetic element (the widespread colonization Island) in the gamma-proteobacteria. I then consider how incongruence tests may be used as a starting point for phylogenetic analysis that accounts for horizontal transfer and duplication events as explanations for homoplasy. PMID- 16243007 TI - Evidence for the existence of two distinct species: Psammomys obesus and Psammomys vexillaris within the sand rats (Rodentia, Gerbillinae), reservoirs of cutaneous leishmaniasis in Tunisia. AB - A thorough taxonomic knowledge about putative animal reservoirs of transmissible diseases is an absolute prerequisite to any ecological investigation and epidemiological survey of zoonoses. Indeed, accurate identification of these reservoirs is essential for predicting species-specific population outbreaks and therefore to develop accurate ecological control strategies. The systematic status of sand rats (genus Psammomys) remains unclear despite the pivotal role of these rodents in the epidemiology of Zoonotic Cutaneous Leishmaniasis (ZCL) disease as sand rats are the main known reservoir hosts of the protozoan parasite Leishmania major. In the present work, we expose morphological, biochemical, genetic and cytogenetic evidence supporting the identification of at least two cryptic species within the genus Psammomys in Tunisia. First, significant morphometric differences were observed and were correlated associated with external features and biogeographic origins. Second, differences in patterns of two isoenzymic systems (Glutamate Oxaloacetate Transaminase (GOT) and 6 PhosphoGluconate Dehydrogenase (6PGD)) were found, which makes it possible to amount these isoenzyme characters to two diagnostic loci. Third, based on the mitochondrial cytochrome b (cyt b) gene, a high magnitude of genetic distance (13.89%) was also observed. Fourth, cytogenetic analysis showed that these two populations groups differ in their diploid chromosome numbers, i.e. 2N=46 versus 2N=48. We consider that all these variations are enough important to be considered as demonstrative and we propose that these two lineages should be considered as two distinct species that we refer to the fat sand rat Psammomys obesus Cretzschmar, 1828 and the thin sand rat Psammomys vexillaris Thomas, 1925. Implications of such results on the eco-epidemiology of ZCL in Tunisia are discussed. PMID- 16243009 TI - Extra-hepatic hepatocellular carcinoma presenting as obstructive jaundice. AB - Hepatocellular carcinoma is a neoplasm with a uniformly poor prognosis. Risk factors for its development include chronic hepatitis B or C infection, haemochromatosis and alpha-1-antitrypsin deficiency, but individuals with any type of chronic liver disease are predisposed. The incidence is significantly higher in Asia and Africa although it has been noted to be increasing in the United States. We present a patient with notable atypical clinical features for hepatocellular carcinoma. The patient had neither predisposing risk factors nor a primary liver lesion causing obstructive jaundice. After multiple tissue specimens were obtained, the final pathological diagnosis was established. Hepatocellular carcinoma generally requires a surgical cure, but patients who are icteric often portend poorer prognoses. For those at high risk, screening may be indicated to identify early curative treatment. PMID- 16243008 TI - Voriconazole therapy in children with cystic fibrosis. AB - BACKGROUND: There is increasing evidence for the efficacy of the antifungal voriconazole, particularly in immunosuppression. We describe our experience of using voriconazole in children with CF. METHODS: We performed a retrospective case note review of children with CF treated with voriconazole in a single centre over an 18 month period. RESULTS: A total of 21 children aged 5 to 16 years (median 11.3) received voriconazole for between 1 and 50 (22) weeks. Voriconazole was used as monotherapy in 2 children with recurrent allergic bronchopulmonary aspergillosis (ABPA); significant and sustained improvements in clinical and serological parameters for up to 13 months were observed, without recourse to oral steroids. Voriconazole was used in combination with an immunomodulatory agent in a further 11 children with ABPA, with significant improvement in pulmonary function and serology. 8 children without ABPA but who had recurrent Aspergillus fumigatus isolates and increased symptoms also received voriconazole; this group did not improve with treatment. Adverse effects occurred in 7 children (33%: photosensitivity reaction 3, nausea 2, rise in hepatic enzymes 1, hair loss 1). CONCLUSIONS: Voriconazole may be a useful adjunctive therapy for ABPA in CF. Voriconazole monotherapy appears to be an alternative treatment strategy when oral corticosteroids may not be suitable. PMID- 16243010 TI - Overt gastrointestinal bleeding in haematologic neoplasms. AB - BACKGROUND AND AIM: Patients with acute leukaemia suffer from various haemorrhages, most frequently due to thrombocytopenia. We could not reach any information regarding the frequency of gastrointestinal bleeding in acute leukaemia and decided to search this complication in patients with acute and chronic leukaemias and myeloproliferative disorders, retrospectively. PATIENTS AND METHODS: During a 6-year period, 291 patients with acute leukaemia, 52 patients with chronic leukaemia and 108 patients with myeloproliferative disorders had been followed. Thirty-two cases of overt gastrointestinal haemorrhage episodes (25 upper, 7 lower) were observed during the mentioned period. RESULTS: The frequency of bleeding episodes was 7.1% (32/451) in haematologic malignancies as a whole, 5.8% (17/291) for acute leukaemia, 1.9% (1/52) for chronic leukaemia and 13% (14/108) for myeloproliferative disorders. If the patients with myeloproliferative disorders in blastic phase were analysed separately, the ratio was 30% (6/20). Oesophagogastroduodenoscopy, which could be performed in 8 of 25 upper gastrointestinal haemorrhage episodes, revealed erosive gastritis in five patients and duodenal ulcers in three patients. Neutropenic enterocolitis was the underlying cause in all of the seven patients with lower gastrointestinal haemorhage. Five out of the seven patients had acute leukaemia. In 7 bleeding attacks, out of 32, the ultimate result was death. Generally, the haemorrhage was only a contributing cause of mortality. All of the mortality cases were patients with acute leukaemia. CONCLUSION: Especially, the patients with myeloproliferative disorders are prone to develop gastrointestinal haemorrhage. The manifestation is generally as upper gastrointestinal bleeding due to gastric erosions and duodenal ulcers. Lower gastrointestinal bleeding is frequently a problem of the patients with acute leukaemia. It is commonly a sign of neutropenic enterocolitis. PMID- 16243011 TI - The quality of life in patients with chronic pancreatitis evaluated using the SF 12 questionnaire: a comparative study with the SF-36 questionnaire. AB - BACKGROUND: In clinical practice there is the need to utilise a time saving questionnaire to assess the quality of life. AIMS: To establish the validity of the SF-12 questionnaire in chronic pancreatitis patients and to identify the predictors capable of modifying the physical and mental summaries in these patients. QUESTIONNAIRES: SF-12 and SF-36 questionnaires were used. SUBJECTS: One hundred and forty-one outpatients with proven chronic pancreatitis. The data of 141 sex- and age-matched Italian subjects of two normative groups (61,434 Italian subjects for SF-12 and 2031 Italian subjects for SF-36) were used as controls. RESULTS: Chronic pancreatitis patients had the SF-12 physical and mental component summaries significantly related to the SF-36 physical and mental component summaries (P<0.001). The presence of pancreatic pain and non-pancreatic surgery accounted for 41.3% in the formation of the PCS-36 score and 37.2% in that of the PCS-12 score, respectively. Gender, BMI and pancreatic pain accounted for 15.3% of the information in the formation of the MCS-36 and for 14.7% in that of the MCS-12; using these clinical variables, the loss of information in applying the SF-12 instead of the SF-36 was very low (4.1 and 0.6% for the PCS and the MCS, respectively). CONCLUSIONS: The SF-12 is a good alternative to the SF-36 in assessing the quality of life in chronic pancreatitis. PMID- 16243012 TI - A large inflammatory fibroid polyp of the colon treated by endoclip-assisted endoscopic polypectomy: A case report. AB - Inflammatory fibroid polyp is a rare benign polypoid lesion of the gastrointestinal tract. Histologically, inflammatory fibroid polyp is characterised by an admixture of numerous small vessels, fibroblasts and oedematous connective tissue, accompanied by marked inflammatory infiltration by eosinophils. A 40-year-old man visited our hospital for the purpose of colorectal screening due to a positive faecal occult blood test. A pedunculated and reddish polyp was found endoscopically in the ascending colon. The polyp was large but was resected endoscopically without any problems. Histologically, the abnormal tissue of the polyp was located in the submucosal and mucosal layer. Proliferation of spindle cells and infiltration of inflammatory cells, such as plasma cells and eosinophils, were observed. Immunohistochemically, the spindle cells were positive for CD34, which was localised in the cytoplasm. These cells were also positive for S100 protein but were negative for c-kit and muscle markers. These findings are compatible with the histological diagnosis of inflammatory fibroid polyp. The surgical margin of the polyp was free of the tumour. Inflammatory fibroid polyp is more commonly found in the stomach or small intestine, and rarely in the colon, and therefore our case is a rare example of large and pedunculated colonic inflammatory fibroid polyp, which was treated successfully by endoscopic polypectomy. PMID- 16243013 TI - Hypoplastic left heart syndrome: from in-utero diagnosis to school age. AB - HLHS can be treated with successful survival outcome. Prenatal diagnosis of the anomaly is now quite common. Our understanding of the developmental aspects of HLHS during the second and third trimesters of gestation is advancing. Survivors of surgery are being closely followed and studied as they proceed forwards in time. A number of morbidities are identified. Many questions concerning the pathophysiological mechanisms of these morbidities exist. New therapies and treatments will certainly arise to meet the challenges these children face as they enter into adulthood, and as our understanding of this unique cardiovascular state progresses. PMID- 16243014 TI - Characteristics of periodontal ligament subpopulations obtained by sequential enzymatic digestion of rat molar periodontal ligament. AB - Periodontal ligament (PDL) consists of different cell populations in various differentiation stages. In the present study, we isolated cell populations from rat molar PDL by sequential enzymatic digestion and characterized growth potential and mineralization activity of the PDL subpopulations (PDL-SP) to throw light on the mechanism of PDL remodeling and, in its turn, periodontal tissue regeneration. PDL attached to extracted rat molars was digested 2 mg/ml collagenase and 0.25% trypsin at 37 degrees C for 30 min. Then four consecutive digestions were performed for 20 min each in a fresh digestive solution. The solutions were centrifuged to collect released cells and 5 PDL subpopulations (30M-, 50M-, 70M-, 90M-and 110M-PDL-SP) were obtained. Light microscopic observation showed that about a half of PDL in width attached on the root surface of extracted teeth and 30M-PDL-SP was considered to contain cells mainly from middle portion of PDL. Scanning electron microscopic examination indicated that 110M-PDL-SP was enriched by root lining cementoblastic cells. 30M-PDL-SP showed a high level of proliferative activity. Although the growth potential of a subpopulation decreased in PDL-SP toward the root surface, 110M-PDL-SP had a high proliferative activity equivalent to that of 30M-PDL-SP. Analyses of alkaline phosphatase (ALP) and mineralization activities showed that higher activities in PDL-SP toward the surface of roots and that 110M-PDL-SP had the highest activity of ALP and the largest number of mineralization nodules. The present study shows as supposed by previous studies on cell kinetics in PDL that subpopulations with larger growth potential were generally located in the middle portion of PDL and those with higher mineralization activities toward the surface of the roots. It is suggested, however, that a possible pathway of PDL cell turnover may exist within the PDL-SP on the root surface in addition to the generally recognized pathway from the middle area of PDL to root surface. PMID- 16243015 TI - Sorting nexins. PMID- 16243016 TI - The chordate ParaHox cluster. PMID- 16243017 TI - Weak suppression of visual context in chronic schizophrenia. PMID- 16243018 TI - Body-size regulation: combining genetics and physiology. AB - New research has revealed that the activity of the insulin-signaling pathway in the prothoracic gland of Drosophila modulates ecdysone release and thereby influences both the duration and rate of larval growth. PMID- 16243019 TI - Genome rearrangements: mother knows best! AB - In Paramecium, developmentally programmed genome rearrangements can be altered by the presence of homologous sequences within the maternal somatic nucleus. Newly identified RNA-binding proteins appear to mediate the transfer of homologous sequence information from the maternal to the developing somatic nucleus, facilitating epigenetic regulation of this large-scale genome reorganization. PMID- 16243020 TI - Photosensing fungi: phytochrome in the spotlight. AB - Red light triggers asexual development and represses sexual development in the fungus Aspergillus nidulans. This response has been shown to require a phytochrome red/far-red light photoreceptor, FphA, which is cytoplasmic and binds a tetrapyrrole chromophore. FphA exhibits similarities to both plant and bacterial phytochromes. PMID- 16243021 TI - Neuroscience: comraderie and nostalgia in nematodes. AB - Two recent papers on social rearing and olfactory imprinting show that early developmental experiences can lead to long-lasting changes in behaviour of the model nematode Caenorhabditis elegans. PMID- 16243022 TI - Counting chromosomes: not as easy as 1, 2, 3. AB - Mammalian cells must count their X chromosomes to determine whether to initiate X chromosome inactivation. A region that may be important for X chromosome counting has been identified, but the puzzle pieces still do not quite fit. PMID- 16243023 TI - Evolution: do bad husbands make good fathers? AB - Males sometimes harm their mates as they seek to maximise the number of offspring they sire. But are females really suffering or do the benefits of having sons that inherit their father's manipulative traits make up for the costs? Three recent studies provide the first hard data addressing this issue, but they differ in their conclusions. PMID- 16243024 TI - Eye movements: building a stable world from glance to glance. AB - Frequent exploratory eye-movements called saccades pose for the visual system the problem of combining information from successive fixations into an apparently seam less conscious experience. A new study shows that information from successive fixations is combined, not by fusing fixation 'snapshots', but by integrating more complex visual attributes at a mid-high level of analysis. PMID- 16243025 TI - Chromosome dynamics: actin's gone fishing. AB - Chromosome congression and segregation have been widely known to be coordinated by the function of the dynamic spindle microtubules. But recent work suggests that oocytes may employ a unique actin-dependent mechanism of chromosome delivery to the spindle. PMID- 16243026 TI - New roles for Galpha and RGS proteins: communication continues despite pulling sisters apart. AB - Large G protein alpha subunits and their attendant regulators of G-protein signaling (RGS) proteins control both intercellular signaling and asymmetric cell divisions by distinct pathways. The classical pathway, found throughout higher eukaryotic organisms, mediates intercellular communication via hormone binding to G-protein-coupled receptors (GPCRs). Recent studies have led to the discovery of GPCR-independent activation of Galpha subunits by the guanine nucleotide exchange factor RIC-8 in both asymmetric cell division and synaptic vesicle priming in metazoan organisms. Protein-protein interactions and protein function in each pathway are driven through the cycle of GTP binding and hydrolysis by the Galpha subunit. This review builds a conceptual framework for understanding RIC-8 mediated pathways by comparison with the mechanism of classical G-protein activation and inhibition in GPCR signaling. PMID- 16243027 TI - Telomere binding protein Taz1 establishes Swi6 heterochromatin independently of RNAi at telomeres. AB - BACKGROUND: The telomere is a specialized heterochromatin conserved among eukaryotes. However, it remains unknown how heterochromatin protein 1 (HP1) is recruited to telomeres and how telomere heterochromatin is formed. In fission yeast, the RNAi (RNA interference)-RITS (RNA-induced initiation of transcriptional silencing) pathway initiates heterochromatin formation at the centromeres and the silent mat locus by using common DNA sequences, the dg and dh repeats, as the templates for small interfering RNA (siRNA). RESULTS: We found that telomeric repeats are sufficient for the establishment of Swi6 (a fission yeast HP1 homolog) heterochromatin, and the establishment requires Taz1, a telomere binding protein of the TRF family. Additionally, Swi6 heterochromatin is established by a part of the subtelomere that contains sequences highly homologous to that of the dh repeat, and it is strikingly destabilized by the deletion of both Taz1 and RNAi-RITS. Transcripts from the telomeric dh-homologous region were specifically associated with RITS, and deletion of the telomeric dh homologous region showed the phenotype similar to that of the rnai mutant in terms of the telomeric silencing, indicating that the RNAi-RITS pathway acts at the telomeric dh-homologous region to establish Swi6 heterochromatin. Furthermore, we found that Taz1 establishes Swi6 heterochromatin independently of the telomeric repeats and the RNAi-RITS pathway at the subtelomeres. CONCLUSION: The telomere heterochromatin is regulated by at least two factors: One is Taz1, which is telomere specific, and the other is RNAi-RITS, which is commonly used at the constitutive heterochromatin regions. PMID- 16243028 TI - Activity-driven dendritic remodeling requires microtubule-associated protein 1A. AB - Activity-prompted dendritic remodeling leads to calcium-influx-dependent activation of signaling pathways within minutes and gene transcription within hours. However, dendrite growth continues for days and requires extension and stabilization of the cytoskeleton in nascent processes. In addition to binding microtubules, microtubule-associated proteins (MAPs) associate with the actin cytoskeleton, anchor ion channels and signaling complexes, and modulate synaptic growth. MAP2 is predominantly dendritic. MAP1B is at postsynaptic densities (PSD) and modulates ion channel activity, in addition to affecting axon growth. Less is known about MAP1A, but it is also enriched in dendrites at input locations, including PSDs where MAP1A associates with channel complexes and the calcium sensor caldendrin. MAP1A rescued hearing loss in tubby mice. Here we show that MAP1A becomes enriched in dendrites concurrently with dendritic branching and synapse formation in the developing brain; that synaptic activity is required for establishing mature MAP1A expression levels; and that MAP1A expression is required for activity-dependent growth, branching, and stabilization of the dendritic arbor. PMID- 16243029 TI - Efficient mitosis in human cells lacking poleward microtubule flux. AB - Chromosome segregation relies on the dynamic properties of spindle microtubules (MTs). Poleward MT flux contributes to spindle dynamics through the disassembly of MT minus ends at spindle poles coupled to the continuous poleward transport of spindle MTs. Despite being conserved in metazoan cells, the function of flux remains controversial because flux rates differ widely in different cell types. In meiotic systems, the rate of flux nearly matches that of chromosome movement, but in mitotic systems, flux is significantly slower than chromosome movement. Here, we show that spindles in human mitotic cells depleted of the kinesin-13 proteins Kif2a and MCAK lack detectable flux and that such cells frequently fail to segregate all chromosomes appropriately at anaphase. Elimination of flux reduces poleward chromosome velocity approximately 20%, but does not hinder bipolar spindle assembly, chromosome alignment, or mitotic progression. Thus, mitosis proceeds efficiently in human cells lacking detectable poleward MT flux. These data demonstrate that in human cultured cells, kinetochores are sufficient to effectively power chromosome movement, leading us to speculate that flux is maintained in these cells to fulfill other functional roles such as error correction or kinetochore regulation. PMID- 16243030 TI - The Aspergillus nidulans phytochrome FphA represses sexual development in red light. AB - Phytochrome photoreceptors sense red and far-red light through photointerconversion between two stable conformations, a process mediated by a linear tetrapyrrole chromophore. Originally, phytochromes were thought to be confined to photosynthetic organisms including cyanobacteria, but they have been recently discovered in heterotrophic bacteria and fungi, where little is known about their functions. It was shown previously in the ascomycetous fungus Aspergillus nidulans that asexual sporulation is stimulated and sexual development repressed by red light. The effect was reminiscent of a phytochrome response, and indeed phytochrome-like proteins were detected in several fungal genomes. All fungal homologs are more similar to bacterial than plant phytochromes and have multifunctional domains where the phytochrome region and histidine kinase domain are combined in a single protein with a C-terminal response-regulator domain. Here, we show that the A. nidulans phytochrome FphA binds a biliverdin chromophore, acts as a red-light sensor, and represses sexual development under red-light conditions. FphA-GFP is cytoplasmic and excluded from the nuclei, suggesting that red-light photoperception occurs in the cytoplasm. This is the first phytochrome experimentally characterized outside the plant and bacterial kingdoms and the second type of fungal protein identified that functions in photoperception. PMID- 16243031 TI - Hypomorphic mutation of PDK1 suppresses tumorigenesis in PTEN(+/-) mice. AB - Many cancers possess elevated levels of PtdIns(3,4,5)P(3), the second messenger that induces activation of the protein kinases PKB/Akt and S6K and thereby stimulates cell proliferation, growth, and survival. The importance of this pathway in tumorigenesis has been highlighted by the finding that PTEN, the lipid phosphatase that breaks down PtdIns(3,4,5)P(3) to PtdIns(4,5)P(2), is frequently mutated in human cancer. Cells lacking PTEN possess elevated levels of PtdIns(3,4,5)P(3), PKB, and S6K activity and heterozygous PTEN(+/-) mice develop a variety of tumors. Knockout of PKBalpha in PTEN-deficient cells reduces aggressive growth and promotes apoptosis, whereas treatment of PTEN(+/-) mice with rapamycin, an inhibitor of the activation of S6K, reduces neoplasia. We explored the importance of PDK1, the protein kinase that activates PKB and S6K, in mediating tumorigenesis caused by the deletion of PTEN. We demonstrate that reducing the expression of PDK1 in PTEN(+/-) mice, markedly protects these animals from developing a wide range of tumors. Our findings provide genetic evidence that PDK1 is a key effector in mediating neoplasia resulting from loss of PTEN and also validate PDK1 as a promising anticancer target for the prevention of tumors that possess elevated PKB and S6K activity. PMID- 16243032 TI - Quantitative analysis of protein dynamics during asymmetric cell division. AB - In dividing Drosophila sensory organ precursor (SOP) cells, the fate determinant Numb and its associated adaptor protein Pon localize asymmetrically and segregate into the anterior daughter cell, where Numb influences cell fate by repressing Notch signaling. Asymmetric localization of both proteins requires the protein kinase aPKC and its substrate Lethal (2) giant larvae (Lgl). Because both Numb and Pon localization require actin and myosin, lateral transport along the cell cortex has been proposed as a possible mechanism for their asymmetric distribution. Here, we use quantitative live analysis of GFP-Pon and Numb-GFP fluorescence and fluorescence recovery after photobleaching (FRAP) to characterize the dynamics of Numb and Pon localization during SOP division. We demonstrate that Numb and Pon rapidly exchange between a cytoplasmic pool and the cell cortex and that preferential recruitment from the cytoplasm is responsible for their asymmetric distribution during mitosis. Expression of a constitutively active form of aPKC impairs membrane recruitment of GFP-Pon. This defect can be rescued by coexpression of nonphosphorylatable Lgl, indicating that Lgl is the main target of aPKC. We propose that a high-affinity binding site is asymmetrically distributed by aPKC and Lgl and is responsible for asymmetric localization of cell-fate determinants during mitosis. PMID- 16243033 TI - Will travel for food: spatial discounting in two new world monkeys. AB - Nonhuman animals steeply discount the future, showing a preference for small, immediate over large, delayed rewards. Currently unclear is whether discounting functions depend on context. Here, we examine the effects of spatial context on discounting in cotton-top tamarins (Saguinus oedipus) and common marmosets (Callithrix jacchus), species known to differ in temporal discounting. We presented subjects with a choice between small, nearby rewards and large, distant rewards. Tamarins traveled farther for the large reward than marmosets, attending to the ratio of reward differences rather than their absolute values. This species difference contrasts with performance on a temporal task in which marmosets waited longer than tamarins for the large reward. These comparative data indicate that context influences choice behavior, with the strongest effect seen in marmosets who discounted more steeply over space than over time. These findings parallel details of each species' feeding ecology. Tamarins range over large distances and feed primarily on insects, which requires using quick, impulsive action. Marmosets range over shorter distances than tamarins and feed primarily on tree exudates, a clumped resource that requires patience to wait for sap to exude. These results show that discounting functions are context specific, shaped by a history of ecological pressures. PMID- 16243034 TI - PDGFRalphaalpha signaling is regulated through the primary cilium in fibroblasts. AB - Recent findings show that cilia are sensory organelles that display specific receptors and ion channels, which transmit signals from the extracellular environment via the cilium to the cell to control tissue homeostasis and function. Agenesis of primary cilia or mislocation of ciliary signal components affects human pathologies, such as polycystic kidney disease and disorders associated with Bardet-Biedl syndrome. Primary cilia are essential for hedgehog ligand-induced signaling cascade regulating growth and patterning. Here, we show that the primary cilium in fibroblasts plays a critical role in growth control via platelet-derived growth factor receptor alpha (PDGFRalpha), which localizes to the primary cilium during growth arrest in NIH3T3 cells and primary cultures of mouse embryonic fibroblasts. Ligand-dependent activation of PDGFRalphaalpha is followed by activation of Akt and the Mek1/2-Erk1/2 pathways, with Mek1/2 being phosphorylated within the cilium and at the basal body. Fibroblasts derived from Tg737(orpk) mutants fail to form normal cilia and to upregulate the level of PDGFRalpha; PDGF-AA fails to activate PDGFRalphaalpha and the Mek1/2-Erk1/2 pathway. Signaling through PDGFRbeta, which localizes to the plasma membrane, is maintained at comparable levels in wild-type and mutant cells. We propose that ciliary PDGFRalphaalpha signaling is linked to tissue homeostasis and to mitogenic signaling pathways. PMID- 16243035 TI - P-Rex1 regulates neutrophil function. AB - Rac GTPases regulate cytoskeletal structure, gene expression, and reactive oxygen species (ROS) production. Rac2-deficient neutrophils cannot chemotax, produce ROS, or degranulate upon G protein-coupled receptor (GPCR) activation. Deficiency in PI3Kgamma, an upstream regulator of Rac, causes a similar phenotype. P-Rex1, a guanine-nucleotide exchange factor (GEF) for Rac, is believed to link GPCRs and PI3Kgamma to Rac-dependent neutrophil responses. We have investigated the functional importance of P-Rex1 by generating a P-Rex1(-/-) mouse. P-Rex1(-/-) mice are viable and healthy, with apparently normal leukocyte development, but with mild neutrophilia. In neutrophils from P-Rex1(-/-) mice, GPCR-dependent Rac2 activation is impaired, whereas Rac1 activation is less compromised. GPCR dependent ROS formation is absent in lipopolysaccharide (LPS)-primed P-Rex1(-/-) neutrophils, but less affected in unprimed or TNFalpha-primed cells. Recruitment of P-Rex1(-/-) neutrophils to inflammatory sites is impaired. Surprisingly, chemotaxis of isolated neutrophils is only slightly reduced, with a mild defect in cell speed, but normal polarization and directionality. Secretion of azurophil granules is unaffected. In conclusion, P-Rex1 is an important regulator of neutrophil function by mediating a subset of Rac-dependent neutrophil responses. However, P-Rex1 is not an essential regulator of neutrophil chemotaxis and degranulation. PMID- 16243036 TI - P-Rex1 is a primary Rac2 guanine nucleotide exchange factor in mouse neutrophils. AB - Leukocyte chemoattractants regulate many leukocyte functions, including leukocyte chemotaxis, via the Rho family of small GTPases that include RhoA, Cdc42, and Rac. Previous work has revealed mechanisms by which chemoattractants regulate RhoA and Cdc42 in mouse neutrophils, but the mechanisms for regulation of Rac remain unclear even though Rac is important for neutrophil functions. Here, we characterized P-Rex1, a Gbetagamma and PIP(3)-regulated guanine nucleotide exchange factor that was initially identified as a Rac activator in response to chemoattractants, for its roles in the regulation of Rac activity and neutrophil functions. We generated a mouse line in which the P-Rex1 gene is disrupted and found that P-Rex1 deficiency did not significantly affect Rac1 activation but diminished Rac2 activation in response to a chemoattractant fMLP in mouse neutrophils. This preference for Rac2 may partially result from the apparent higher affinity of P-Rex1 for Rac2 than for Rac1 because P-Rex1 was more readily immunoprecipitated with Rac2(S17N) than Rac1(S17N). In addition, P-Rex1 deficiency significantly attenuated fMLP-induced F actin formation and superoxide production without affecting LPS- or PMA-induced production. Furthermore, P-Rex1 deficiency caused a chemotactic defect that is primarily attributed to a reduction in the migration rate rather than directionality. PMID- 16243040 TI - Reading the literature and developing a hypothesis. PMID- 16243037 TI - DNA-PKcs-dependent signaling of DNA damage in Dictyostelium discoideum. AB - DNA double-strand breaks (DSBs) can be repaired by either homologous recombination (HR) or nonhomologous end-joining (NHEJ). In vertebrates, the first step in NHEJ is recruitment of the DNA-dependent protein kinase (DNA-PK) to DNA termini. DNA-PK consists of a catalytic subunit (DNA-PKcs) that is recruited to DNA ends by the Ku70/Ku80 heterodimer. Although Ku has been identified in a wide variety of organisms, to date DNA-PKcs has only been identified experimentally in vertebrates. Here, we report the identification of DNA-PK in the nonvertebrate Dictyostelium. Dictyostelium Ku80 contains a conserved domain previously implicated in recruiting DNA-PKcs to DNA and consistent with this observation, we have identified DNA-PKcs in the Dictyostelium genome. Disruption of the gene encoding Dictyostelium DNA-PKcs results in sensitivity to DNA DSBs and defective H2AX phosphorylation in response to this form of DNA damage. However, these phenotypes are only apparent when DNA damage is administered in G(1) phase of the cell cycle. These data illustrate a cell cycle-dependent requirement for Dictyostelium DNA-PK in signaling and combating DNA DSBs and represent the first experimental verification of DNA-PKcs in a nonvertebrate organism. PMID- 16243041 TI - Fundamentals of surgical research course: research presentations. AB - BACKGROUND: The effective communication of research outcomes is a skill that can be learned. To make the greatest impact, it is imperative to know your audience. METHODS: Oral communication is a highly effective means for dissemination of new information. The preparation of a research presentation requires significant time. The researcher must determine the key messages to be delivered and plan the presentation accordingly. High quality visual aids can impress your audience and, in combination with a succinct oral presentation, can increase retention of the material. CONCLUSIONS: In addition to the communication of the raw scientific data and results, the importance of stance, appearance, voice, and eye contact necessary to engage your audience should not be overlooked. PMID- 16243044 TI - Research ethics primer. PMID- 16243045 TI - Developing the young academic surgeon. AB - In the past, the process of developing the young academic surgeon was arguably less strategic, one that was often not deliberately managed and monitored, leading in some cases to academic drift and disillusionment. Once upon a time it was assumed that greatness was genetic and that the next triple threat would emerge when a pre-programmed set of genes was turned on. Today, as the complexities and vicissitudes of our work increase, it is practically impossible for even the most gifted young person to be successful without careful attention to career development. Faculty development must be deliberate and strategic- every junior faculty member is unique and will require a customized career development plan that is well thought out, linked to measurable goals, monitored routinely and buttressed by effective mentoring. This approach will require time and commitment--precious commodities that are in short supply as the demands on our time are only escalating. By recruiting the right people (those who fit with the organization's values and goals) and providing the right environment, we can optimize the growth and satisfaction of our young faculty and, in so doing, create departments that are leaders in carrying out our missions of research, education and patient care. We cannot afford to have our young people fail--it is simply too costly, both from a financial and a human perspective. PMID- 16243046 TI - New measurement of hepatic blood flow by xenon CT system: an animal study with PGE1. AB - BACKGROUND: A new Xenon computed tomography (CT) system was developed to measure both hepatic arterial and portal venous tissue blood flow (HATBF/PVTBF) non invasively. Despite its clinical trial, the effect of prostaglandin E1 (PGE1) on hepatic hemodynamics is not well investigated. In a rabbit model, we evaluated the accuracy of this system by comparing it with the electromagnetic blood flowmeter (EMBF), the pharmacological effect of PGE1 on the fractional hepatic hemodynamics. MATERIALS AND METHODS: Seven NZW-rabbits were used. Serial abdominal CT scan was obtained every min before and during the 4 min inhalation of the Xenon gas, followed by 5 min administration of oxygen air. From these images, HATBF and PVTBF were separately calculated with a special new imaging system. We also used EMBF during laparotomy, and directly measured the hepatic arterial and portal venous flow with or without PGE1 administration. RESULTS: Xenon CT showed HATBF of 18.4 +/- 4.5 (ml/min/100 g) and PVTBF of 69.4 +/- 15.0, was almost identical with those of EMBF (19.8 +/- 5.7 and 67.2 +/- 19.1, respectively). After PGE1 administration, Xenon CT showed 22.9 +/- 4.6 and 76.5 +/- 20.5, while those with EMBF were 21.0 +/- 6.5 and 84.7 +/- 21.6, respectively. There were significant correlations (P < 0.01) in total HTBF, HATBF, and PVTBF between results of Xenon CT and EMBF. CONCLUSIONS: Xenon CT with a newly developed imaging system enables us to measure the fractional hepatic tissue blood flow in rabbits, differentially and accurately. Venous administration of PGE1 increased total hepatic blood flow, mainly affecting the portal blood flow. PMID- 16243047 TI - The effect of N-acetylcysteine on pulmonary lipid peroxidation and tissue damage. AB - BACKGROUND: We aimed to investigate the effect of N-acetylcysteine (NAC) on pulmonary lipid peroxidation and tissue damage in experimental obstructive jaundice (OJ) stimulated by lipopolysaccharide (LPS) in this study. MATERIALS AND METHODS: We randomized 40 rats into five groups. Group A: Sham (n = 8); group B: OJ (n = 8); group C: OJ + lipopolysaccharide (LPS; n = 8); group D: OJ + NAC + LPS (n = 8); group E: OJ + LPS + NAC (n = 8). OJ was performed by common bile duct ligation and division in all groups except the sham group. At the fifth day, the rats were jaundiced. At the fifth day of OJ, LPS was injected 10 mg/kg intraperitoneally to the rats and at the tenth day, the rats were sacrificed in group C. In group D; at the fifth day of OJ, NAC was started 100 mg/kg subcutaneously and the same dose NAC injection repeated every day for 5 days. At the tenth day of OJ, LPS was injected 10 mg/kg intraperitoneally to the rats and then after 6 h they were sacrificed. In group E; 10 mg/kg LPS was administered intraperitoneally at fifth day of OJ and after then NAC was started 100 mg/kg subcutaneously and the same dose NAC injection repeated every day for 5 days and at the tenth day, the rats were sacrificed. Tissue samples were harvested through a midline incision, and lungs were resected and examined histopathologically and immunohistochemically for tissue damage scoring. The blood was taken by cardiac puncture and malondialdehyde (MDA), myeloperoxidase (MPO), and levels of total antioxidant status were detected with biochemical methods to evaluate lung tissue damage. RESULTS: Increase in lung and serum MDA and MPO levels, as well as decrease in total antioxidant status, were observed in groups B and C when compared with the sham group (P = 0.0001, for each comparison). Furthermore, the lung tissue damage was observed in the same groups by histopathological examination when compared with sham group. There was significant decrease at serum and lung MPO and MDA levels after the NAC application in groups D and E, when compared with group C (P = 0.0001, for each comparison). Antioxidant status in groups D and E were increased in the presence of NAC (P = 0.0001, for each comparison). Lung histology was prevented relatively in group D when compared with groups B and C. CONCLUSION: Results of the study indicate that NAC has protective effect on pulmonary lipid peroxidation and tissue damage before and after LPS administration. PMID- 16243048 TI - Heat shock response inhibits NF-kappaB activation and cytokine production in murine Kupffer cells. AB - BACKGROUND: Kupffer cells play a crucial role in the pathogenesis of sepsis through production of proinflammatory mediators and control of systemic endotoxemia. The anti-inflammatory effects of heat shock response (HSP) have been well documented. However, the role of HSP in lipopolysaccharide (LPS) induced Kupffer cell activation has not been fully investigated. In this study, we investigated the effects of HSP on LPS induced Kupffer cell NF-kappaB activation and cytokine production. MATERIALS AND METHODS: Kupffer cells were isolated from mice by collagenase digestion and HSP was induced by culturing Kupffer cells with sodium arsenite. Kupffer cells were stimulated in vitro by LPS. Heat shock protein (HSP)-70 expression and cytoplasmic IkappaBalpha protein was determined by Western blot. Supernatant tumor necrosis factor (TNF)-alpha, interleukin (IL) 6 and IL-10 levels were measured by ELISA. NF-kappaB activation was analyzed by electrophoresis mobility shift assay. Cytokine and IkappaBalpha mRNA expression were determined by RT-PCR. Toll-like receptor 4 expression on Kupffer cells was determined by flow cytometry. RESULTS: HSP pre-conditioning significantly inhibited LPS-induced cytokine TNF-alpha and IL-6 production and mRNA expression. NF-kappaB activation and IkappaBalpha degradation induced by LPS were attenuated by HSP. HSP up-regulated expression of IkappaBalpha mRNA. No effect of HSP on cell surface expression of TLR4 was observed. CONCLUSIONS: Increased IkappaBalpha stability and up-regulation of IkappaBalpha gene expression may be one of the mechanisms of the inhibition of LPS induced Kupffer cell activation by HSP. HSP also inhibited expression of the anti-inflammatory cytokine IL-10, and the mechanism and biological significance of this effect merit further investigation. PMID- 16243049 TI - Dopamine dependency of cognitive switching and response repetition effects in Parkinson's patients. AB - A group of people with Parkinson's disease and a group of matched controls were tested on a task involving a switch between perceptual dimensions. Patients were tested both 'on' and 'off' their normal medication cycles. Stimuli appeared in pairs for each trial, with each stimulus consisting of a color and a shape. One dimension of color and one of shape were mapped to each of two response keys. A cue was presented concurrently with each stimulus to indicate whether to respond on the basis of color or shape, following procedures developed by Hayes et al. [Hayes, A.E., Davidson, M.C., Keele, S.W., and Rafal, R.D. (1998). Toward a functional analysis of the basal ganglia. Journal of Cognitive Neuroscience, 10, 178-198]. Replicating previous literature, abnormally large switch costs were observed in patients who were off their normal medication cycles. A novel finding was that patients in the 'on' state demonstrated a slight reversal of switch costs. Also novel, reaction time (RT) costs associated with switching between response keys, and interactions between response switching and task switching were influenced predominantly by on-off dopamine manipulations. It is concluded that abnormal task switching costs and response repetition effects likely reflect impairments of activation and inhibition, and both effects are dopamine dependent. PMID- 16243050 TI - Exploring the functional architecture of person recognition system with event related potentials in a within- and cross-domain self-priming of faces. AB - In this paper, we explored the functional properties of person recognition system by investigating the onset, magnitude, and scalp distribution of within- and cross-domain self-priming effects on event-related potentials (ERPs). Recognition of degraded pictures of famous people was enhanced by a prior exposure to the same person's face (within-domain self-priming) or name (cross-domain self priming) as compared to those preceded by neutral or unrelated primes. The ERP results showed first that the amplitude of the N170 component to famous face targets was modulated by within- and cross-domain self-priming, suggesting not only that the N170 component can be affected by top-down influences but also that this top-down effect crosses domains. Second, similar to our behavioral data, later ERPs to famous faces showed larger ERP self-priming effects in the within domain than in the cross-domain condition. In addition, the present data dissociated between two topographically and temporally overlapping priming sensitive ERP components: the first one, with a strongly posterior distribution arising at an early onset, was modulated more by within-domain priming irrespective whether the repeated face was familiar or not. The second component, with a relatively uniform scalp distribution, was modulated by within- and cross domain priming of familiar faces. Moreover, there was no evidence for ERP-induced modulations for unfamiliar face targets in the cross-domain condition. Together, our findings suggest that multiple neurocognitive events that are possibly mediated by distinct brain loci contribute to face priming effects. PMID- 16243051 TI - A functional MRI study of preparatory signals for spatial location and objects. AB - We investigated preparatory signals for spatial location and objects in normal observers using functional magnetic resonance imaging (fMRI). Activity for attention-directing cues was separated from activity for subsequent test arrays containing the target stimulus. Subjects were more accurate in discriminating a target face among distracters when they knew in advance its location (spatial directional cue), as compared to when the target could randomly appear at one of two locations (spatial neutral cue), indicating that the spatial cue was used. Spatially specific activations occurred in a region at the intersection of the ventral intraparietal sulcus and transverse occipital sulcus (vIPS-TOS), which showed significantly stronger activation for rightward- than leftward-directing cues, while other fronto-parietal areas were activated by the cue but did not show spatial specificity. In visual cortex, activity was weak or absent in retinotopic occipital regions following attention-directing cues and this activity was not spatially specific. In a separate task, subject discriminated a target outdoor scene among distracters after the presentation of spatial neutral cues. There was no significant difference in dorsal frontoparietal activity during the face versus scene discrimination task. Also, there was only weak evidence for selective preparatory activity in ventral object-selective regions, although the activation of these regions to the subsequent test array did depend upon which discrimination (face or place) was performed. We conclude first that under certain circumstances, spatial cues that produce strong behavioral effects may modulate parietal-occipital regions in a spatially specific manner without producing similar modulations in retinotopic occipital regions. Second, attentional modulations of object-selective regions in temporal-occipital cortex can occur even though preparatory object-selective modulations of those regions are absent or weak. PMID- 16243052 TI - Behavioral and neurobiological effects of printed word repetition in lexical decision and naming. AB - A series of experiments studied the effects of repetition of printed words on (1) lexical decision (LD) and naming (NAM) behavior and (2) concomitant brain activation. It was hypothesized that subword phonological analysis (assembly) would decrease with increasing word familiarity and the greater decrease would occur in LD, a task that is believed to be less dependent on assembly than naming. As a behavioral marker of assembly, we utilized the regularity effect (the difference in response latency between words with regular versus irregular spelling-sound correspondences). In addition to repetition, stimulus familiarity was manipulated by word frequency and case alternation. Both experiments revealed an initial latency disadvantage for low frequency irregular words suggesting that assembly is the dominant process in both tasks when items are unfamiliar. As items become more familiar with repetition, the regularity effect disappeared in LD but persisted in NAM. Brain activation patterns for repeated words that were observed in fMRI paralleled the behavioral studies in showing greater reductions in activity under lexical decision than naming for regions previously identified as involved in assembly. PMID- 16243053 TI - The role of the right hemisphere in processing nonsalient metaphorical meanings: application of principal components analysis to fMRI data. AB - Some researches indicate that the right hemisphere (RH) has a unique role in comprehending the figurative meaning of metaphors whereas the results of other studies do not support the notion of a selective role for the RH in accessing metaphorical meanings. The present research used fMRI technology to test a theoretical explanation of the above conflicting findings. This theoretical account is derived from the Graded Salience Hypothesis (GSH) [Giora, R. (1997). Understanding figurative and literal language: the Graded Salience Hypothesis. Cognitive Linguistics, 7, 183-206; Giora, R. (2003). On our mind: Salience, context and figurative language. New York: Oxford University Press], according to which the degree of meaning salience, rather than literality or nonliterality primarily affects differences between the LH and RH in linguistic processing. Thus, the GSH predicts a selective RH involvement in comprehension of novel, nonsalient metaphoric meanings and LH involvement in the comprehension of conventional, salient metaphoric meanings. Fifteen normal adults participated in a block designed fMRI experiment that compared the patterns of brain activation induced by processing the meanings of literal, conventional metaphoric, novel metaphoric and unrelated word pairs. The subjects performed a semantic judgment task. We applied the Principal Components Analysis (PCA) technique in order to find different functional networks corresponding to the different stimuli. Our results, obtained from PCA of the fMRI data indicate that the right homologue of Wernicke's area has a special role in processing novel metaphors. We suggest that a unique network, consisting of the right homologue of Wernicke's area, right and left premotor areas, right and left insula and Broca's area, is recruited for the processing of novel metaphors but not for the processing of conventional metaphors. PMID- 16243054 TI - Similar cortical correlates underlie visual object identification and orientation judgment. AB - Visual object perception has been suggested to follow two different routes in the human brain: a ventral, view-invariant occipital-temporal route processes object identity, whereas a dorsal, view-dependent occipital-parietal route processes spatial properties of an object. Using fMRI, we addressed the question whether these routes are exclusively involved in either object recognition or spatial representation. We presented subjects with images of natural objects and involved them either in object identification or object orientation judgment task. For both tasks, we observed activation in ventro-temporal as well as parietal areas bilaterally, with significantly stronger responses for the orientation judgment in both ventro-temporal as well as parietal areas. Our findings suggest that object identification and orientation judgment do not follow strictly separable cortical pathways, but rather involve both the dorsal and the ventral stream. PMID- 16243055 TI - Material-specific long-term memory representations of faces and spatial positions: evidence from slow event-related brain potentials. AB - Motivated by models that propose material-specific cortical long-term memory representations we expected different topographies of event-related slow waves of the EEG during cued retrieval of two distinct types of information (faces and spatial positions), which are assumed to be processed and stored in topographically distinct cortical areas, i.e., in either the ventral or the dorsal visual pathway. Seventeen participants learned associations either between words and spatial positions or between words and faces. Each word was associated with either one or two positions or faces. In a cued recall test, one day later, participants saw two words and had to decide whether these were linked to each other via an associated spatial position or a face. Slow event-related potentials (ERPs) of the EEG were recorded from 61 scalp electrodes during both acquisition and recall. Response times increased monotonically with the number of faces and positions to be reactivated. Negative slow ERPs showed a comparable topography during anticipation learning and cued recall, but dissociated topographically for positions and faces. The maximum of the negativity increased when items were presented repetitively (compared to the first presentation) during learning, and also with the number of the to-be-reactivated associations during retrieval. These results are consistent with an information-processing model that assumes material-specific cortical representations of episodic memory contents, which are established as localized cortical cell assemblies during encoding, and which are being reactivated during recall. PMID- 16243056 TI - Normal and abnormal face selectivity of the M170 response in developmental prosopagnosics. AB - Developmental prosopagnosia is a lifelong impairment in face recognition despite normal low-level visual processing. Here we used magnetoencephalography (MEG) to examine the M170 response, a component occurring approximately 170 ms after stimulus onset, in a group of five developmental prosopagnosics. In normal subjects, the M170 is "face-selective", with a consistently higher amplitude to faces than to a wide variety of other visual stimulus categories; the N170, a component recorded using event-related potentials (ERP) and thought to be analogous to the M170, also shows this "face selectivity". Two previous ERP studies with developmental prosopagnosics have found attenuation or absence of face selectivity in the N170 response of these subjects [Bentin, S., Deouell, L. Y., and Soroker, N. (1999). Selective visual streaming in face recognition: Evidence from developmental prosopagnosia. Neuroreport, 10, 823-827; Kress, T., and Daum, I. (2003). Event-related potentials reflect impaired face recognition in patients with congenital prosopagnosia. Neuroscience Letters, 352, 133-136]. Three of our developmental prosopagnosic group showed this non-selective pattern at the M170 while the remaining two prosopagnosics were indistinguishable from normal controls. Thus, impaired face recognition is not necessarily correlated with an absence of the "face-selective" M170. Furthermore, ERP recordings collected simultaneously in the two developmental prosopagnosics with seemingly selective M170s also showed N170s within the same normal selective range, demonstrating that the face-selective signals found with MEG are not due to differences between MEG and ERP. While the presence of face selectivity at these neurophysiological markers is insufficient for predicting normal behavioral performance with faces, it could help to distinguish different classes of face recognition deficits. PMID- 16243058 TI - Recent Advances in Non-viral Gene Delivery. AB - Gene therapy has been deemed the medicine of the future due to its potential to treat many types of diseases. However, many obstacles remain before gene delivery is optimized to specific target cells. Over the last several decades, many approaches to gene delivery have been closely examined. By understanding the factors that determine the efficiency of gene uptake and expression as well as those that influence the toxicity of the vector, we are better able to develop new vector systems. This chapter will provide a brief overview of recent advances in gene delivery, specifically on the development of novel non-viral vectors. The following chapters will provide additional details regarding the evolution of non viral gene delivery systems. PMID- 16243059 TI - Barriers to Gene Delivery Using Synthetic Vectors. AB - Progress has been made in the development of different types of nucleic acids such as DNA and siRNA with the potential to form the basis of new treatments for genetic and acquired disorders. The lack of suitable vectors for the delivery of nucleic acids, however, represents a major hurdle to their continued development and therapeutic application. Synthetic vectors based on polycations are promising vectors for gene delivery as they are relatively safe and can be modified by the incorporation of ligands for targeting to specific cell types. However, the levels of gene expression mediated by synthetic vectors are low compared to viral vectors. The aim of this chapter is to give an overview of the main barriers that have been identified as limiting gene transfer using polycation-based synthetic vectors. The chapter is divided into two sections to focus on both extracellular and intracellular barriers. We describe novel strategies that are being used to develop increasingly sophisticated vectors in an attempt to overcome these barriers. For instance, we describe approaches to prolong the plasma circulation of polyplexes by the incorporation onto their surface of hydrophilic polymers such as polyethylene glycol (PEG) and poly[N-(2-hydroxypropyl)methacrylamide] (pHPMA). In addition, strategies to improve transfer of nucleic acids from the outside of the cell to the nucleus are described to overcome barriers such as escape from endocytic vesicles and translocation across the nuclear membrane. Furthermore, we highlight new types of vectors based on reducible polycations that are triggered by the intracellular environment to facilitate efficient cytoplasmic release of nucleic acids. PMID- 16243060 TI - Pharmacokinetics of Plasmid DNA-Based Non-viral Gene Medicine. AB - Non-viral gene therapy can be realized by optimization of the pharmacokinetic properties of both the vector and the encoded therapeutic protein. A major obstacle to its successful clinical application is the limited ability of plasmid DNA, the most convenient gene-coding compound, to distribute within the body after in vivo administration. Under normal conditions, plasmid DNA and its non viral vector complexes have difficulty in passing through various anatomical and biological barriers. These characteristics greatly limit the number and distribution of cells transduced with the vector, because transgene expression only occurs in cells that are reached by the vector. New approaches to the design of vectors as well as the methods of administration, such as electroporation and a hydrodynamic delivery, have increased the transgene expression in vivo, suggesting that improved distribution of plasmid DNA is possible by these approaches. In this chapter, the basic pharmacokinetic properties of naked plasmid DNA under normal conditions are first reviewed, then the properties of both naked and complexed plasmid DNA are discussed under conditions where significant transgene expression takes place. PMID- 16243061 TI - What Role Can Chemistry Play in Cationic Liposome-Based Gene Therapy Research Today? AB - Gene therapy research is still in trouble owing to a paucity of acceptable vector systems to deliver nucleic acids to patients for therapy. Viral vectors are efficient but may be too dangerous for routine clinical use. Synthetic non-viral vectors are inherently much safer but are currently not efficient enough to be clinically viable. The solution for gene therapy lies with improved synthetic non viral vectors based upon well-found platform technologies and a thorough understanding of the barriers to efficient gene delivery and expression (transfection) relevant to clinical applications of interest. Here we introduce and interpret synthetic non-viral vector systems through the ABCD nanoparticle structural paradigm that represents, in our view, an appropriate lens through which to view all synthetic, non-viral vector systems applicable to in vitro use or in vivo applications and gene therapy. Our intention in introducing this paradigm is to shift the focus of organic and physical chemists away from the design of yet another cytofectin, and instead encourage them to appreciate the wider challenges presented by the need to produce tool kits of meaningful chemical components from which to assemble viable, tailor-made nanoparticles for in vivo applications and gene therapy, both now and in the future. PMID- 16243062 TI - Lipoplex Structures and Their Distinct Cellular Pathways. AB - Cationic liposomes (CLs) are used as non-viral vectors in worldwide clinical trials of gene therapy. Among other advantages, CL-DNA complexes have the ability to transfer very large genes into cells. However, since the understanding of their mechanisms of action is still incomplete, their transfection efficiencies remain low compared to those of viruses. We describe recent studies which have started to unravel the relationship between the distinct structures and physicochemical properties of CL-DNA complexes and their transfection efficiency by combining several techniques: synchrotron X-ray diffraction for structure determination, laser-scanning confocal microscopy to probe the interactions of CL DNA particles with cells, and luciferase reporter-gene expression assays to measure transfection efficiencies in mammalian cells. Most CL-DNA complexes form a multilayered structure with DNA sandwiched between the cationic lipids (lamellar complexes, L(alpha)(C)). Much more rarely, an inverted hexagonal structure (H(II)(C)) with single DNA strands encapsulated in lipid tubules is observed. An important recent insight is that the membrane charge density sigma(M) of the CL-vector, rather than, for example, the charge of the cationic lipid, is a universal parameter governing the transfection efficiency of L(alpha)(C) complexes. This has led to a new model of the intracellular release of L(alpha)(C) complexes, through activated fusion with endosomal membranes. In contrast to L(alpha)(C) complexes, H(II)(C) complexes exhibit no dependence on sigma(M), since their structure leads to a distinctly different mechanism of cell entry. Surface-functionalized complexes with poly(ethyleneglycol)-lipids (PEG lipids), potentially suitable for transfection in vivo, have also been investigated, and the novel aspects of these complexes are discussed. PMID- 16243063 TI - "Diffusible-PEG-Lipid Stabilized Plasmid Lipid Particles" AB - Many viral and non-viral gene transfer systems suffer from common pharmacological issues that limit their utility in a systemic context. By application of the liposomal drug delivery paradigm, many of the limitations of the first generation non-viral delivery systems can be overcome. Encapsulation in small, long circulating particles called stabilized plasmid lipid particles (SPLP) results in enhanced accumulation at disease sites and selective protein expression. This work compares the detergent dialysis method of SPLP manufacture with an alternative method, spontaneous vesicle formation by ethanol dilution. The pharmacology of SPLP, as determined by monitoring lipid label and quantitative real time PCR, is also presented. PMID- 16243064 TI - Toxicity of Cationic Lipid-DNA Complexes. AB - As with any conventional drug, the body's response to cationic lipid-DNA complexes is highly dependent on both the dose administered and the route of delivery. At relatively low doses there is little to no effect on organ function or tissue architecture, but at higher doses, acute inflammation and tissue damage can occur that is sometimes quite profound. Of the two most common routes of delivery, intravenous (IV) or intrapulmonary, IV administration tends to cause more severe adverse effects and can be lethal at higher doses of complex. Both routes activate an innate immune response that includes the induction of proinflammatory cytokines and immune cell activation, a major portion of which has been attributed to the presence of immunostimulatory CpG motifs within the plasmid DNA vector. Removing CpGs from the plasmid vector reduces several, but not all of the acute inflammatory responses to cationic lipid-DNA complexes. Therefore, other strategies are required to improve the therapeutic potential of these vectors, such as transient immune suppression, aerosolization of the complex, and novel formulations that have increased efficiency of transduction and decreased interaction with immune cells. PMID- 16243065 TI - Polyethylenimine (PEI). AB - Since the first edition of this book in 1999 the field of gene therapy has been the arena both for major advances that justified the early hopes placed in the concept, and for ever-present impatience with the slowness of overall progress. On the positive side, gene therapy obtained its first brilliant success, though not where most efforts were invested and not with a synthetic vector (Cavazzana Calvo et al., 2000). Yet the search for efficient molecules is still very active, in part because the negative consequences of using viral vectors somewhat shadow the brilliant picture (Hacein-Bey-Abina et al., 2003). PMID- 16243066 TI - Pluronic Block Copolymers for Gene Delivery. AB - Amphiphilic block copolymers of poly(ethylene oxide) and poly(propylene oxide) called Pluronic or poloxamer are commercially available pharmaceutical excipients. They recently attracted considerable attention in gene delivery applications. First, they were shown to increase the transfection with adenovirus and lentivirus vectors. Second, they were shown to increase expression of genes delivered into cells using non-viral vectors. Third, the conjugates of Pluronic with polycations, were used as DNA-condensing agents to form polyplexes. Finally, it was demonstrated that they can increase regional expression of the naked DNA after its injection in the skeletal and cardiac muscles or tumor. Therefore, there is substantial evidence that Pluronic block copolymers can improve gene expression with different delivery routes and different types of vectors, including naked DNA. These results and possible mechanisms of Pluronic effects are discussed. At least in some cases, Pluronic can act as biological adjuvants by activating selected signaling pathways, such as NF-kappaB, and upregulating the transcription of the genes. PMID- 16243067 TI - Terplex Gene Delivery System. AB - Polymeric gene delivery systems have been developed to overcome problems caused by viral carriers. They are low cytotoxic, have no size limit, are convenient in handling, of low cost and reproducible. A Terplex gene delivery system consisting of plasmid DNA, low density lipoprotein and hydropholized poly-L-lysine was designed and characterized. The plasmid DNA, when formulated with stearyl PLL and LDL, forms a stable and hydrophobicity/charge-balanced Terplex system of optimal size for efficient cellular uptake. DNA is still intact after the Terplex formation. This information is expected to be utilized for the development of improved transfection vector for in vivo gene therapy. Terplex DNA complex showed significantly longer retention in the vascular space than naked DNA. This system was used in the augmentation of myocardial transfection at an infarction site with the VEGF gene. PMID- 16243068 TI - Design of Polyphosphoester-DNA Nanoparticles for Non-Viral Gene Delivery. AB - Development of safe and effective non-viral gene carriers is still critical to the ultimate success of gene therapy. This review highlights our attempt to design the gene carriers in a systematic manner. We have synthesized a series of polymers with a phosphoester backbone containing different charge groups in the sidechain connected to the backbone through a phosphate (PO) or a phosphoramide (PN) bond. These gene carriers have different charge groups, sidechain lengths, and branching structures, but they are structurally related to allow a systematic investigation of the structure-property relationship, including DNA binding capacity, cytotoxicity, DNA protection, biodegradability, DNA release kinetics, and transfection efficiency. PMID- 16243069 TI - Development of HVJ Envelope Vector and Its Application to Gene Therapy. AB - To create a highly efficient vector system that is minimally invasive, we initially developed liposomes that contained fusion proteins from the hemagglutinating virus of Japan (HVJ; Sendai virus). These HVJ-liposomes delivered genes and drugs to cultured cells and tissues. To simplify the vector system and develop more efficient vectors, the next approach was to convert viruses to non-viral vectors. Based on this concept, we recently developed the HVJ envelope vector. HVJ with robust fusion activity was inactivated, and exogenous DNA was incorporated into the viral envelope by detergent treatment and centrifugation. The resulting HVJ envelope vector introduced plasmid DNA efficiently and rapidly into both cultured cells in vitro and organs in vivo. Furthermore, proteins, synthetic oligonucleotides, and drugs have also been effectively introduced into cells using the HVJ envelope vector. The HVJ envelope vector is a promising tool for both ex vivo and in vivo gene therapy experiments. Hearing impairment in rats was prevented and treated by hepatocyte growth factor gene transfer to cerebrospinal fluid using HVJ envelope vector. For cancer treatment, tumor-associated antigen genes were delivered efficiently to mouse dendritic cells to evoke an anti-cancer immune response. HVJ envelope vector fused dendritic cells and tumor cells and simultaneously delivered cytokine genes, such as IL-12, to the hybrid cells. This strategy successfully prevented and treated cancers in mice by stimulating the presentation of tumor antigens and the maturation of T cells. For human gene therapy, a pilot plant to commercially produce clinical grade HVJ envelope vector has been established. PMID- 16243070 TI - Targeting of Polyplexes: Toward Synthetic Virus Vector Systems. AB - Dominating issues in gene vector optimization are specific in recognizing the target cells and exploiting the proper intracellular trafficking routes. Any progress in this area will result in improved specific gene transfer, reduce the required therapeutic vector doses and, in consequence, lower the overall toxicity to the host. To provide polyplexes with the ability to distinguish between non target and target cells, cell-binding ligands have been incorporated which recognize target-specific cellular receptors. In addition, polyplex domains with unspecific binding capacity (such as surface charges) have to be shielded or removed. Cell-binding ligands can be small molecules, vitamins, carbohydrates, peptides or proteins such as growth factors or antibodies. Such ligands have been incorporated into polyplexes after chemical conjugation to cationic polymers. The choice of the ligand and physical properties of the DNA formulation strongly influence extracellular routing (circulation in blood, tissue distribution), uptake and intracellular delivery of polyplexes. Recent efforts are discussed that aim at the development of polyplexes into virus-like supramolecular complexes; such particles should undergo structural changes compatible with extracellular and intracellular targeting. PMID- 16243071 TI - Germany's need for health-care reforms. PMID- 16243072 TI - Wellbeing: an idea whose time has come. PMID- 16243073 TI - Boosting the effectiveness of food aid. PMID- 16243074 TI - Voriconazole for candidosis: an important addition? PMID- 16243075 TI - BCG: the story continues. PMID- 16243077 TI - Research and clinical ethics after the tsunami: Sri Lanka. PMID- 16243076 TI - Reorienting health-research communication. PMID- 16243078 TI - Deep brain stimulation for treatment of refractory depression. PMID- 16243079 TI - Nobel Prize winners Robin Warren and Barry Marshall. PMID- 16243080 TI - Antibiotics for acute infective conjunctivitis in children. PMID- 16243081 TI - Antibiotics for acute infective conjunctivitis in children. PMID- 16243084 TI - Antibiotics for acute infective conjunctivitis in children. PMID- 16243085 TI - Assisted dying for the terminally ill. PMID- 16243086 TI - Sir Richard Doll. PMID- 16243087 TI - Hiroshima. PMID- 16243088 TI - Voriconazole versus a regimen of amphotericin B followed by fluconazole for candidaemia in non-neutropenic patients: a randomised non-inferiority trial. AB - BACKGROUND: Voriconazole has proven efficacy against invasive aspergillosis and oesophageal candidiasis. This multicentre, randomised, non-inferiority study compared voriconazole with a regimen of amphotericin B followed by fluconazole for the treatment of candidaemia in non-neutropenic patients. METHODS: Non neutropenic patients with a positive blood culture for a species of candida and clinical evidence of infection were enrolled. Patients were randomly assigned, in a 2:1 ratio, either voriconazole (n=283) or amphotericin B followed by fluconazole (n=139). The primary efficacy analysis was based on clinical and mycological response 12 weeks after the end of treatment, assessed by an independent data-review committee unaware of treatment assignment. FINDINGS: Of 422 patients randomised, 370 were included in the modified intention-to-treat population. Voriconazole was non-inferior to amphotericin B/fluconazole in the primary efficacy analysis, with successful outcomes in 41% of patients in both treatment groups (95% CI for difference -10.6% to 10.6%). At the last evaluable assessment, outcome was successful in 162 (65%) patients assigned voriconazole and 87 (71%) assigned amphotericin B/fluconazole (p=0.25). Voriconazole cleared blood cultures as quickly as amphotericin B/fluconazole (median time to negative blood culture, 2.0 days). Treatment discontinuations due to all-cause adverse events were more frequent in the voriconazole group, although most discontinuations were due to non-drug-related events and there were significantly fewer serious adverse events and cases of renal toxicity than in the amphotericin B/fluconazole group. INTERPRETATION: Voriconazole was as effective as the regimen of amphotericin B followed by fluconazole in the treatment of candidaemia in non neutropenic patients, and with fewer toxic effects. RELEVANCE TO PRACTICE: There are several options for treatment of candidaemia in non-neutropenic patients, including amphotericin B, fluconazole, voriconazole, and echinocandins. Voriconazole can be given both as initial intravenous treatment and as an oral stepdown agent. PMID- 16243089 TI - Effect of BCG vaccination on risk of Mycobacterium tuberculosis infection in children with household tuberculosis contact: a prospective community-based study. AB - BACKGROUND: Little is known about factors that affect the risk of acquiring infection in children exposed to Mycobacterium tuberculosis. The effect of BCG vaccination has been difficult to ascertain because the tuberculin skin test (TST), until recently the only method for detecting M tuberculosis infection, does not reliably distinguish between tuberculosis infection and BCG vaccination. METHODS: We investigated risk factors for tuberculosis infection in 979 child household contacts of 414 adult index patients with sputum smear-positive pulmonary tuberculosis in Istanbul, Turkey. Children were aged up to 16 years (median 7, IQR 3-11) and 770 of 979 (79%) had a BCG scar. A T-cell-based enzyme linked immunospot assay (ELISpot), which is not confounded by BCG vaccination, and TST were used to assess infection. Independent risk factors for infection were identified through multivariate analysis. FINDINGS: Amount of tuberculosis exposure within the household and age (a marker of tuberculosis exposure outside the household) were strongly associated with likelihood of infection as measured by both TST and ELISpot. ELISpot also identified absence of BCG scar as an independent risk factor for infection in tuberculosis-exposed children; BCG vaccinated children had an odds ratio of 0.60 (95% CI 0.43-0.83, p=0.003) for tuberculosis infection, compared with unvaccinated children. INTERPRETATION: Contrary to the prevailing theory that BCG vaccination protects only against tuberculosis disease, our results suggest that the vaccine also protects against tuberculosis infection. This finding has important implications for our understanding of the biology of tuberculosis infection and development of improved tuberculosis vaccines. PMID- 16243091 TI - Co-coverage of preventive interventions and implications for child-survival strategies: evidence from national surveys. AB - BACKGROUND: In most low-income countries, several child-survival interventions are being implemented. We assessed how these interventions are clustered at the level of the individual child. METHODS: We analysed data from Bangladesh, Benin, Brazil, Cambodia, Eritrea, Haiti, Malawi, Nepal, and Nicaragua. A co-coverage score was obtained by adding the number of interventions received by each child (including BCG, diphtheria-pertussis-tetanus, and measles vaccines), tetanus toxoid for the mother, vitamin A supplementation, antenatal care, skilled delivery, and safe water. Socioeconomic status was assessed through principal components analysis of household assets, and concentration indices were calculated. FINDINGS: The percentage of children who did not receive a single intervention ranged from 0.3% (14/5495) in Nicaragua to 18.8% (1154/6144) in Cambodia. The proportions receiving all available interventions varied from 0.8% (48/6144) in Cambodia to 13.3% (733/5495) in Nicaragua. There were substantial inequities within all countries. In the poorest wealth quintile, 31% of Cambodian children received no interventions and 17% only one intervention; in Haiti, these figures were 15% and 17%, respectively. Inequities were inversely related to coverage levels. Countries with higher coverage rates tended to show bottom inequity patterns, with the poorest lagging behind all other groups, whereas low coverage countries showed top inequities with the rich substantially above the rest. INTERPRETATION: The inequitable clustering of interventions at the level of the child raises the possibility that the introduction of new technologies might primarily benefit children who are already covered by existing interventions. Packaging several interventions through a single delivery strategy, while making economic sense, could contribute to increased inequities unless population coverage is very high. Co-coverage analyses of child-health surveys provide a way to assess these issues. PMID- 16243090 TI - Differences between organophosphorus insecticides in human self-poisoning: a prospective cohort study. AB - BACKGROUND: Although more than 100 organophosphorus insecticides exist, organophosphorus poisoning is usually regarded as a single entity, distinguished only by the compound's lethal dose in animals. We aimed to determine whether the three most common organophosphorus insecticides used for self-poisoning in Sri Lanka differ in the clinical features and severity of poisoning they cause. METHODS: We prospectively studied 802 patients with chlorpyrifos, dimethoate, or fenthion self-poisoning admitted to three hospitals. Blood cholinesterase activity and insecticide concentration were measured to determine the compound and the patients' response to insecticide and therapy. We recorded clinical outcomes for each patient. FINDINGS: Compared with chlorpyrifos (35 of 439, 8.0%), the proportion dying was significantly higher with dimethoate (61 of 264, 23.1%, odds ratio [OR] 3.5, 95% CI 2.2-5.4) or fenthion (16 of 99, 16.2%, OR 2.2, 1.2-4.2), as was the proportion requiring endotracheal intubation (66 of 439 for chlorpyrifos, 15.0%; 93 of 264 for dimethoate, 35.2%, OR 3.1, 2.1-4.4; 31 of 99 for fenthion, 31.3%, 2.6, 1.6-4.2). Dimethoate-poisoned patients died sooner than those ingesting other pesticides and often from hypotensive shock. Fenthion poisoning initially caused few symptoms but many patients subsequently required intubation. Acetylcholinesterase inhibited by fenthion or dimethoate responded poorly to pralidoxime treatment compared with chlorpyrifos-inhibited acetylcholinesterase. INTERPRETATION: Organophosphorus insecticide poisoning is not a single entity, with substantial variability in clinical course, response to oximes, and outcome. Animal toxicity does not predict human toxicity since, although chlorpyrifos is generally the most toxic in rats, it is least toxic in people. Each organophosphorus insecticide should be considered as an individual poison and, consequently, patients might benefit from management protocols developed for particular organophosphorus insecticides. PMID- 16243092 TI - Haemoglobin E beta thalassaemia in Sri Lanka. AB - Haemoglobin E beta thalassaemia is the commonest form of severe thalassaemia in many Asian countries, but little is known about its natural history, the reasons for clinical diversity, or its management. We studied 109 Sri Lankan patients with the disorder over 5 years. 25 patients were not receiving transfusion; transfusion was stopped with no deleterious effect in a further 37. We identified several genetic and environmental factors that might contribute to the phenotypic diversity of the disorder, including modifiers of haemoglobin F production, malaria, and age-related changes in adaptive function. Our findings suggest that haemoglobin E beta thalassaemia can be managed without transfusion in many patients, even with low haemoglobin levels. Age-related changes in the pattern of adaptation to anaemia suggest that different and more cost-effective approaches to management should be explored. PMID- 16243093 TI - Self-harm. AB - The term self-harm is commonly used to describe a wide range of behaviours and intentions including attempted hanging, impulsive self-poisoning, and superficial cutting in response to intolerable tension. As with suicide, rates of self-harm vary greatly between countries. 5-9% of adolescents in western countries report having self-harmed within the previous year. Risk factors include socioeconomic disadvantage, and psychiatric illness--particularly depression, substance abuse, and anxiety disorders. Cultural aspects of some societies may protect against suicide and self-harm and explain some of the international variation in rates of these events. Risk of repetition of self-harm and of later suicide is high. More than 5% of people who have been seen at a hospital after self-harm will have committed suicide within 9 years. Assessment after self-harm includes careful consideration of the patient's intent and beliefs about the lethality of the method used. Strong suicidal intent, high lethality, precautions against being discovered, and psychiatric illness are indicators of high suicide risk. Management after self-harm includes forming a trusting relationship with the patient, jointly identifying problems, ensuring support is available in a crisis, and treating psychiatric illness vigorously. Family and friends may also provide support. Large-scale studies of treatments for specific subgroups of people who self-harm might help to identify more effective treatments than are currently available. Although risk factors for self-harm are well established, aspects that protect people from engaging in self-harm need to be further explored. PMID- 16243094 TI - Genetic epidemiology and public health: hope, hype, and future prospects. AB - Genetic epidemiology is a rapidly expanding research field, but the implications of findings from such studies for individual or population health are unclear. The use of molecular genetic screening currently has some legitimacy in certain monogenic conditions, but no established value with respect to common complex diseases. Personalised medical care based on molecular genetic testing is also as yet undeveloped for common diseases. Genetic epidemiology can contribute to establishing the causal nature of environmentally modifiable risk factors, through the application of mendelian randomisation approaches and thus contribute to appropriate preventive strategies. Technological and other advances will allow the potential of genetic epidemiology to be revealed over the next few years, and the establishment of large population-based resources for such studies (biobanks) should contribute to this endeavour. PMID- 16243095 TI - HIV prevention research in a resource-limited setting: the experience of planning a trial in Cambodia. PMID- 16243096 TI - Visual loss due to a carious tooth. PMID- 16243097 TI - The association between childhood and adolescent sexual abuse and proxies for sexual risk behavior: a random sample of the general population of Sweden. AB - OBJECTIVE: Several studies with small and "high risk" samples have demonstrated that a history of childhood or adolescent sexual abuse (CASA) is associated with sexual risk behaviors (SRBs). However, few studies with large random samples from the general population have specifically examined the relationship between CASA and SRBs with a comprehensive set of measures. DESIGN: The study was a cross sectional retrospective survey of past and current sexual health and behavior. METHODS: A random sample of 4781 persons from the Swedish Post Address Register was obtained, which included 6,119,000 Swedish citizens in 1996. Of those persons, 2810 participants agreed to participate in the study. Participants were interviewed as well as administered a questionnaire regarding their sexual health and behavior. RESULTS: Using Mann-Whitney U tests, a history of CASA was found to be associated with younger age at first intercourse; younger age at diagnosis of first sexually transmitted infection (STI); greater frequency of unintended pregnancy; greater likelihood of participation in group sex; higher likelihood of not interrupting sexual intercourse despite the risk of pregnancy or risk of an STI; greater likelihood of exchanging sex for money or other necessities/drugs; more frequent substance use in the last 48 hours; and higher likelihood of adult sexual and physical assault. CONCLUSIONS: The findings of this study confirm previous research, which has reported an association between CASA and SRBs in smaller and high-risk samples. Clinicians working with adults with a history of CASA should be aware of the relationship between CASA and SRBs and be prepared to address such issues during therapy. PMID- 16243099 TI - Prevention of high-dose-rate brachytherapy accidents. ICRP Publication 97. AB - High-dose-rate brachytherapy is a rapidly growing technique (HDR) that has been replacing low-dose-rate (LDR) procedures over the last few years in both industrialised and developing countries. It is estimated that about 500,000 procedures (administration of treatment) are performed by HDR units annually. LDR equipment has been discontinued by many manufacturers over the last few years, leaving HDR brachytherapy as the major alternative. HDR brachytherapy techniques deliver a very high dose, of the order of 1.6-5.0 Gy/min, so mistakes can lead to under- or overdosage with the potential for clinical adverse effects. More than 500 HDR accidents (including one death) have been reported along the entire chain of procedures from source packing to delivery of dose. Human error has been the prime cause of radiation events. In the present report, the International Commission on Radiological Protection concludes that many accidents could have been prevented if staff had had functional monitoring equipment and paid attention to the results. Since iridium has relatively short half-life, the HDR sources need to be replaced approximately every 4 months. Over 10,000 HDR sources are transported annually, with the resultant potential for accidents; therefore, appropriate procedures and regulations must be observed. A number of specific recommendations on procedures and equipment are given in this report. The need for an emergency plan and for practising emergency procedures is stressed. The possibility of loss or theft of sources must be kept in mind. A collaborating team of specifically trained personnel following quality assurance (QA) procedures is necessary to prevent accidents. Maintenance is indispensable component of QA; external audits of procedures re-enforce good and safe practice, and identify potential causes of accidents. QA should include peer review of cases. Accidents and incidents should be reported and the lessons learned should be shared with other users to prevent similar mistakes. PMID- 16243102 TI - Arrhythmogenic right ventricular cardiomyopathy in two pairs of monozygotic twins. AB - Arrhythmogenic right ventricular cardiomyopathy (ARVC) is an inheritant disease with an autosomal dominant mode of transmission with incomplete penetrance and variable expression. Linkage analysis in affected families succeeds in identifying 9 loci determining 9 subtypes of the disease. Genotype phenotype correlation is unclear and the influence of various environmental factors is discussed. OBJECTIVES: Genotype phenotype correlation in 2 pairs of monozygotic twins with ARVC and the role of environmental factors are analyzed. PATIENTS AND METHODS: Among 40 pts with ARVC and their 195 relatives there were 2 pairs of monozygotic twins: brothers, age 47 y; and sisters, age 48 y. History, ECG, Holter monitoring, 2D and Doppler Echo, and MRI were analyzed. RESULTS: Twin brothers: ARVC was diagnosed in the proband after the episode of VT with LBBB morphology (enlarged right ventricle, focal hypokinesia of apex, MR evidence of adipose tissue in RV wall). Identical morphology of RV was seen in asymptomatic twin brother. The patient presenting arrhythmia has been rowing for 4 years. Twin sisters: diagnosis was done during family screening. Both were asymptomatic. RV morphology typical for ARVC was found discrete in one of them (bulges adipose tissue in the RV apex); the latter showed changes suggesting RV abnormality (mild segmental dilatation of infundibulum, adipose tissue in a free wall of the RV). No differences in previous viral infections and sports involvement were observed. CONCLUSIONS: 1. Clinical picture of ARVC in monozygotic twins is not identical. 2. Strenuous effort may be a factor triggering the arrhythmia in pts with ARVC. PMID- 16243103 TI - Utility of combined parameters of common carotid intima-media thickness or albuminuria in diagnosis of coronary artery disease in women. AB - BACKGROUND: Noninvasive testing for suspected coronary artery disease is challenging. We prospectively investigated whether measurements of carotid intima media thickness in the presence or absence of albuminuria in patients with stable chest pain syndromes can be used as a noninvasive test algorithm for prediction of significant coronary artery disease. Additionally, this algorithm was tested with regard to gender differences. METHODS: Consecutive patients (79 men and 72 women) with stable chest pain syndromes and suspected coronary artery disease admitted for coronary angiography were studied. Measurements of intima-media thickness were performed by ultrasound. Urinary albumin excretion was measured in a random urine specimen. A positive test for coronary artery disease was defined as an intima-media thickness >or=1 mm or albuminuria. RESULTS: Sensitivity, specificity and positive likelihood ratio for a combination of intima-media thickness values >or=1 mm or presence of albuminuria to predict coronary artery disease were, respectively, 0.5, 0.73 and 1.27 in men and 0.68, 0.79 and 3.32 in women. Sensitivity, specificity and positive likelihood ratio for exercise ECG were, respectively, 0.54, 0.48 and 1.08 in men and 0.47, 0.45 and 0.8 in women. CONCLUSIONS: Intima-media thickness of the common carotid artery or presence of albuminuria are clinically valuable parameters in the noninvasive diagnostic work up of women with stable chest pain syndromes. Their value is limited in men. PMID- 16243104 TI - Trauma associated with acute myocardial infarction in a multi-state hospitalized population. AB - INTRODUCTION: Trauma has been suggested, in case series, as one of the nonatherosclerotic mechanisms leading to acute myocardial infarction (AMI), the leading cause of death in the US. AMI following non-penetrating injury has been shown to carry significant morbidity and mortality. OBJECTIVE: To determine whether hospitalized injuries in a large multi state population are associated with increased risk of AMI during the initial hospital stay. METHODS: Statewide injury hospital discharge data were collected from 19 states in 1997. Affected body regions of interest included thoracic, abdominal or pelvic, spine or back and blunt cardiac injury (BCI). The outcome of interest was AMI which was identified based on ICD-9-CM discharge diagnoses for the same visit. Unadjusted and adjusted multivariate logistic regression analyses were performed. RESULTS: Independent of confounding factors and coronary arteriography (CA) status, BCI was associated with 2.6-fold increased risk for AMI in persons 46 years or older. When the diagnosis of AMI was confirmed by CA, BCI was associated with 8-fold risk elevation among patients 46 years and older and a 31-fold elevation among patients 45 years and younger. Abdominal or pelvic trauma, irrespective of confounding factors and CA status, was associated with a 65% increase in the risk of AMI among patients 45 years and younger and 93% increase in the risk of among patients 46 years and older. When the diagnosis of AMI was confirmed by CA, abdominal or pelvic trauma was associated with 6-fold risk elevation among patients 46 years and older. CONCLUSION: Direct trauma to the heart, as characterized by a diagnosis of BCI, was observed to carry the greatest risk for AMI. Abdominal or pelvic trauma also increased the risk for AMI. Longitudinal studies are warranted to better understand the relationship between trauma and AMI. PMID- 16243105 TI - Is coronary angiography in unstable patients safe in district general hospitals without any on-site revascularisation? AB - BACKGROUND: Patients admitted to hospital with unstable angina or non-ST segment elevation myocardial infarction have a high risk of death, re-infarction or re hospitalisation within the next 6 months. International guidelines recommend an early interventional strategy in moderate- to high-risk patients with pre discharge coronary angiography. In the UK, such patients admitted to district hospitals have traditionally been transferred to tertiary centres for investigation and treatment. Due to the large numbers involved and scarcity of tertiary centre beds, this results in long delays before transfer. The objective of this study was to determine whether diagnostic cardiac catheterisation in moderate- to high-risk patients could be safely performed in hospitals without on site revascularisation and whether this strategy led to earlier discharge times. METHODS: A retrospective audit was undertaken of all patients undergoing inpatient coronary angiography to a large district general hospital (DGH) after admission with a clinical diagnosis of unstable angina or non-ST elevation myocardial infarction over a 12-month period from April 2002 to March 2003. The main outcome measures were complications arising from coronary angiography and number of bed-days saved by allowing earlier discharge. RESULTS: 142 patients with non-ST elevation ACS who met 'high risk' criteria underwent inpatient angiography locally. Significant luminal coronary disease was present in 76% of patients but 49% were treated medically and discharged early. 32% of patients required transfer for percutaneous intervention (PCI) and 19% were referred directly for surgery. There was a high revascularisation rate (43%) even in patients who did not have elevated troponin levels on admission. No patients died or sustained a myocardial infarction as a result of angiography, and morbidity was minimal. Patients waited an average of 3 days for an angiogram locally, but transfer time to a tertiary centre was 9 days for PCI and 12 days for surgery. As almost half of all patients were discharged without requiring transfer for revascularisation, we estimate a total of 490 bed-days were saved over 12 months. CONCLUSION: Cardiac catheterisation in most 'high-risk' patients with non-ST elevation ACS is safe in DGHs without on-site PCI or surgery and frees up large numbers of acute medical beds by selecting out only those patients requiring onward referral for revascularisation. PMID- 16243107 TI - The effect of resistance training on left ventricular function and structure of patients with chronic heart failure. AB - INTRODUCTION: Resistance training (RT) has been shown to improve chronic heart failure (CHF) patients' functional ability and quality of life. Despite these benefits RT has not always been recommend as a form of exercise principally because of a concern for acceleration of the left ventricular (LV) remodeling process. This study investigated the effects of 8 weeks RT on the LV structure and function of patients with CHF. METHOD: Fifteen men who suffered from CHF were divided into either a RT program or non-training control group. Before and after 8 weeks of training patients underwent resting echocardiography to assess their end-diastolic and end-systolic dimensions (EDD and ESD, respectively), ejection fraction (EF), fractional shortening (FS) and stroke volume (SV). RESULTS: A repeated measured ANOVA showed that 8 weeks of RT had no significant effect on the LV measurements (group x time, p>0.05). Post training comparison, however, revealed that the EF and FS of the training group was significantly higher than in the control group (40.9+/-10.5% vs. 30.3+/-4.6%, p=0.029 and 25.0+/-7.0% vs. 17.4+/-3.1%, p=0.020 respectively). CONCLUSION: RT is a suitable method of training for CHF patients since it does not cause a reduction of LV contractility function or enhance myocardial deterioration as measured by EF and FS. PMID- 16243106 TI - Influence of age, gender, and serum triglycerides on heart rate in a cohort of asymptomatic individuals without heart disease. AB - BACKGROUND: Age, sex and blood lipids were demonstrated in epidemiological studies to influence heart rate measured on physical examination, on 12-lead electrocardiogram or with automatic devices for short-term measurements. We hypothesized that in healthy individuals, age, sex and other clinical variables may also influence heart rate measured on 24-h ambulatory electrocardiographic monitoring. METHODS: We studied 625 asymptomatic individuals with normal clinical examination, aged 15 to 83 (mean 42, standard deviation 11.9) years, 276 (44.2%) men and 349 (55.8%) women. Heart rate was evaluated on 24 h ambulatory electrocardiographic monitoring. Variables selected in univariate analysis (chi(2) and Student t tests) were further submitted to multivariate analysis with canonical correlation to assess the strength of associations between heart rate and other variables, and multiple linear regression models to generate reference curves. RESULTS: Age was the most significant influence on canonical variable of heart rate relative to other clinical and laboratory variables (0.55; p<0.01). There was an increase in the minimum heart rate and a decrease of maximum heart rate with increasing age in both genders. The increase was steeper in men and the decrease was steeper in women. Minimum heart rate increased with increasing serum triglycerides and decreased as estimated maximum oxygen consumption increased. CONCLUSIONS: There was a narrower variation of heart rate with increasing age in both genders in healthy individuals. This variation was less pronounced in women. In addition, status of body haemostasis associated with peculiar metabolic conditions expressed in serum triglycerides levels may also be associated with heart rate. PMID- 16243108 TI - Combined right ventricular systolic and diastolic dysfunction represents a strong determinant of poor prognosis in patients with symptomatic heart failure. AB - BACKGROUND: The presence of right ventricular systolic dysfunction is known to significantly worsen prognosis of patients with heart failure. However, the prognostic impact of right ventricular diastolic dysfunction and of its combination with right ventricular systolic dysfunction and with other prognostic markers has not yet been systematically studied. The aim of this study was to assess the prognostic impact of combined right ventricular systolic and diastolic dysfunction in patients with symptomatic heart failure due to ischemic or idiopathic dilated cardiomyopathy. METHODS: The study included 177 consecutive patients with symptomatic heart failure (mean left ventricular ejection fraction of 23%). All patients underwent clinical and laboratory examination, standard echocardiography completed by Doppler tissue imaging of the tricuspid annular motion, and right-sided heart catheterization. They were followed up for a mean period of 16 months (range, 1-48 months). RESULTS: During the follow-up, there were 28 cardiac-related deaths and 35 non-fatal cardiac events (31 hospitalizations for heart failure decompensation and 4 hospitalizations for malignant arrhythmias requiring the implantation of a cardioverter defibrillator). The multivariate stepwise Cox regression modeling revealed the right ventricular systolic (represented by the peak systolic tricuspid annular velocity-Sa) and diastolic (represented by the peak early diastolic tricuspid annular velocity-Ea) function to be the independent predictors of event-free survival or survival (p<0.01). The Sa separated better between patients with and without the risk of cardiac events (p<0.05), while the Ea appeared to further distinguish patients with increased risk (those at risk of late event from those at risk of early non-fatal event and early death). The strongest predictive information was obtained by the combination of Sa and Ea creating the Sa/Ea categories. The Sa/Ea I category of patients (Sa>or=10.8 cm s(-1) and Ea>or=8.9 cm s(-1)) had excellent prognosis. On the other hand, the Sa/Ea IV category (Sa<10.8 cm s(-1) and Ea<8.9 cm s(-1)) was found to be at a very high risk of cardiac events (p<0.001 vs. Sa/Ea I). Imbalanced categories of patients (Sa/Ea II and III) with only one component (Sa or Ea) pathologically decreased were at medium risk when assessing event-free survival. However, a significantly better survival (p<0.05) was found in patients with Ea>or=8.9 cm s(-1) (Sa/Ea I and III categories) as compared with those having Ea<8.9 cm s(-1) (Sa/Ea II and IV categories). Thus, in contrast to event-free survival, the survival pattern was determined mainly by the Ea value with only little additional contribution of Sa. CONCLUSIONS: The assessment of right ventricular systolic and diastolic function provides complementary information with a very high power to stratify prognosis of patients with heart failure. The combination of right ventricular systolic and diastolic dysfunction identifies those with a very poor prognosis. PMID- 16243109 TI - Acute pulmonary edema complicating myocardial infarction: is colour M-mode Doppler of the mitral valve a valuable tool? AB - OBJECTIVE: To assess the value of colour M-mode Doppler of the mitral valve in patients with acute myocardial infarction complicated by pulmonary edema. DESIGN: Case-control, clinical. SETTING: Coronary care unit in a university hospital. PATIENTS/SUBJECTS: 28 patients admitted because of acute myocardial infarction, and who developed pulmonary edema (group P, cases); 39 patients with uncomplicated myocardial infarction (group C, controls). RESULTS: Patients in group P showed higher E wave (77+/-20 vs. 64+/-16 cm/s, p=0.007), E/A ratio (1.5+/-1.0 vs. 1.0+/-0.4, p=0.014), lower time of deceleration of the E wave (153+/-40 vs. 196+/-53 ms, p=0.001) and lower Ejection Fraction (35+/-10 vs. 49+/ 11, p<0.001). There were no differences in the velocity of the colour M-mode Doppler of the mitral valve (Vp: 36.2+/-11 vs. 34.0+/-12 cm/s). Excluding patients with abnormal relaxation or restrictive pattern of the pulsed Doppler, Vp was identical (group P 34+/-10, group C 34+/-12). CONCLUSIONS: M-mode colour Doppler of the mitral valve was not useful to differentiate patients with acute pulmonary edema complicating myocardial infarction. Measurement of Vp is not warranted as a routine in these patients. PMID- 16243110 TI - Tissue Doppler echocardiographic evidence of atrial mechanical dysfunction in coronary artery disease. AB - BACKGROUND: Atrial function is an integral part of cardiac function which is often neglected. The presence of coronary artery disease (CAD) may impair atrial function. This study investigated if atrial mechanical dysfunction was present in patients with CAD by tissue Doppler echocardiography (TDI). METHODS: Echocardiography with TDI was performed in 118 patients with CAD, and compared with 100 normal controls with comparable age and heart rate. Regional atrial function was assessed at the left (LA) and right (RA) atrial free wall and inter atrial septum (IAS). The peak regional atrial contraction velocity of (V(A)) and the timing of mechanical events were compared. RESULTS: The V(A) in the LA (5.0+/ 2.6 Vs 7.7+/-2.6 cm/s), IAS (4.8+/-1.7 Vs 5.7+/-1.5 cm/s) and RA (6.8+/-3.1 Vs 9.2+/-2.9 cm/s) were significantly decreased in patients with CAD when compared with controls (all p<0.001). Patients with impaired systolic function (ejection fraction50% (both p<0.001); and were lower in those with restrictive filling pattern (RFP) than non-RFP of diastolic dysfunction (both p<0.05). The V(A) in all the subgroups was lower than controls. In contrast, transmitral atrial velocity was unable to reveal any abnormality except in the subgroup with a RFP. The LA dimension, area and volume were increased in the disease groups, but were largely unchanged in the RA despite abnormal V(A). The physiological inter-atrial delay for the onset and peak atrial contraction between the RA and LA were unaffected by CAD. CONCLUSIONS: The atrial contractile function in both atria was impaired in the presence of CAD, especially in the LA. This was detected even in patients with preserved systolic function or mild diastolic dysfunction such as non-RFP. Direct assessment of atrial velocity by TDI may better reflect atrial mechanical function than transmitral atrial velocity. PMID- 16243111 TI - Long-term follow-up after invasive approach of coronary artery disease in daily practice. AB - BACKGROUND: To assess long-term survival in unselected patients with coronary artery disease in who an invasive approach is considered. METHODS: All patients with significant coronary artery disease who were presented for coronary revascularisation to two tertiary centres in 1992 were included. Follow-up data were collected in September 2002. Multivariate Cox' proportional-hazards regression analysis was applied to assess the independent relation between variables and 10-year survival. RESULTS: A total of 877 patients were included in this analysis. Mean age was 62 and the most common clinical diagnosis was chronic stable angina (60%). Diabetes was present in 12% of the patients. During the follow-up period, 233 patients (27%) died. Predictors of long-term survival were increasing age, diabetes, peripheral vascular disease and a decreased left ventricular function. Compared to medical treated patients, those treated with revascularisation (either by PCI or CABG) had a decreased long-term mortality (p<0.05). Of the patients with PCI 27% had died, compared to 24% in those who had CABG and 36% of those who were treated medically. However, after adjusting for differences in baseline variables, conservative treatment was no significant predictor of long-term mortality. After multivariable analyses, increasing age, decreased left ventricular function and diabetes were independent predictors of long-term mortality. CONCLUSIONS: In patients with coronary artery disease in whom an invasive approach is considered, increasing age, impaired left ventricular function and diabetes are the strongest predictors of long-term mortality. After adjustments for differences in baseline variables, invasive treatment is not associated with a lower long-term mortality. PMID- 16243113 TI - Design of PREVENCION: a population-based study of cardiovascular disease in Peru. AB - Latin America is undergoing the epidemiologic transition that occurred earlier in developed countries, and is likely to face a gigantic epidemic of heart disease in the next few years unless urgent action is taken. The first essential component of any effective cardiovascular disease (CVD) control program is to establish reliable estimates of cardiovascular disease-related morbidity and mortality. However, such data from population-based studies in Latin America are still lacking. In this paper, we present the design and operation of PREVENCION (Estudio Peruano de Prevalencia de Enfermedades Cardiovasculares, for Peruvian Study of the Prevalence of Cardiovascular diseases). PREVENCION is an ongoing population-based study on a representative sample of the civilian non institutionalized population of the second largest city in Peru. Its population is comparable to the rest of the Peruvian urban population and closely resembles other Latin American populations in countries such as Bolivia and Ecuador. Our study will contribute to the enormous task of understanding and preventing CVD in Latin America. PMID- 16243112 TI - Expression changes of thrombospondin-1 and neuropeptide Y in myocardium of STZ induced rats. AB - Diabetic cardiomyopathy was the most dangerous diabetic complication facing diabetics, with its exact mechanisms remaining obscure. Our study was conducted to investigate the expression of thrombospondin-1 (TSP-1) and neuropeptide Y (NPY) in myocardium of streptozotocin (STZ)-induced diabetic rats. We employed streptozotocin (STZ)-induced diabetic rats to study the alteration of the TSP-1 and NPY expression in the left ventricle myocardium in diabetic and normal group by immunohistochemistry and immunofluorescence. The data of weight, blood sugar and urine sugar indicated no significant difference between the two groups before the animal model was induced. Four weeks after the induction of diabetes the weight of the diabeteic animals was 189.1+/-18.4 g, plasma glucose was 23.7+/ 3.25 mmol/L and urine glucose was (++) to (+++); whereas the weight of the control animals was 260.5+/-32.1 g, plasma glucose was 4.9+/-0.5 mmol/L and urine glucose undetectable (-). The differences between the control and the diabetes group were distinct. A significant increase of the TSP-1 and NPY expression was also observed in the diabetic rat's heart. The number of the NPY positive myocardium and the light density of the positive myocardium in the left ventricle of the diabetic model were 17.3+/-2.1 and 102.5+/-9.3/mm(2), respectively, which were considered as increased when compared with the control that were 10.1+/-2.6 and 61.2+/-6.7, respectively. Our results support the view that high glucose conditions can induce an increased synthesis of TSP-1 through the PKC-TGF-beta TSP-1 pathway, which in turn facilitate TGF-beta activation. Additionally, the activation of PKC may further lead to the over-expression of NPY. This may be involved in diabetic cardiomyopathy. PMID- 16243114 TI - Circulating level of gelatinase activity predicts ventricular remodeling in patients with acute myocardial infarction. AB - BACKGROUND: Matrix metalloproteinase (MMP) plays an important role in the development of ventricular remodeling in an animal model of acute myocardial infarction (AMI). We examined whether circulating MMP activity can predict left ventricular (LV) remodeling after AMI in humans. METHODS: We measured the circulating level of MMP-2 and MMP-9 activities (gelatinase activity) at 14 days after the onset of AMI by gelatin zymography in 52 consecutive patients (age 62+/ 2). All patients underwent direct PTCA and stenting at an acute stage, and were treated subsequently with losartan or enalapril. Biplane left ventriculography was performed at admission, and 2 weeks and 6 months after the onset of AMI. RESULTS: We expressed gelatinolysis activity as the ratio to MMP-2 standard. Mean gelatinase activity was 0.721+/-0.013. We divided patients into two groups, groups with gelatinolysis activity <0.72 (low group, n=27) and >0.72 (high group, n=25). Either change in LV end-diastolic volume index (LVEDVI, ml/m(2)) or end systolic volume index (LVESVI, ml/m(2)) from admission to 2 weeks was not different between the two groups. Changes in both LVEDVI and LVESVI from 2 weeks to 6 months were greater in high gelatinolysis activity group than those in low activity group. Moreover, circulating level of gelatinolysis activity was positively correlated with changes in LVEDVI and LVESVI from 2 weeks to 6 months. CONCLUSION: These results demonstrate that circulating level of gelatinase activity can predict LV remodeling after AMI. Inhibition of gelatinase activity at the acute phase may be a therapeutic strategy for the prevention of remodeling after AMI. PMID- 16243116 TI - Association of 22q11 deletion with isolated congenital heart disease in three Chinese ethnic groups. AB - BACKGROUND: Congenital heart disease (CHD) is the most common type of heart disease among children. About 75% of DiGeorge syndrome (DGS) and velo-cardio facial syndrome (VCFS) includes CHD. A deletion within chromosome 22q11.2 has been identified in the majority of patients with DGS and VCFS. And 22q11.2 deletion has become one of the markers used to study CHD in these syndromes. Whether 22q11.2 deletion is associated with isolated CHD is not known and was the topic of this study. METHODS AND RESULTS: We studied the 22q11.2 deletion in three Chinese ethnic groups (Tai, Bai and Han people) with 19 sporadic, isolated CHD by genotype and haplotype analysis with D22S420 etc. 11 consecutive polymorphic microsatellite markers. Among 19 isolated CHD patients, four had Tetralogy of Fallot (TOF), five exhibited Ventricular Septal Defect (VSD), five showed Atrial Septal Defect (ASD) and 5 had Patent Ductus Arteriosus (PDA). In some isolated CHD patients, 3 Mb and 1.5 Mb deletion to chromosome 22q11.2 was found. 2 of 4 TOF (50%) and 1 of 5 VSD (20%) and 1 of 5 PDA (20%) respectively were found to have deletions at D22S944. CONCLUSIONS: 22q11.2 deletion can be detected in isolated TOF, VSD and PDA of three Chinese ethnic groups, without detectable 22q11.2 deletion in those isolated ASD patients examined thus far. Our finding may be the first to show the 22q11.2 deletion in sporadic, isolated PDA/VSD patients whose family members are without CHD. In addition, D22S420 etc. 11 consecutive polymorphic microsatellite markers are very useful for the determination of 22q11.2 deletion in isolated CHD in China. PMID- 16243115 TI - Effects of resveratrol on vascular tone and endothelial function of human saphenous vein and internal mammary artery. AB - BACKGROUND: The polyphenolic compound resveratrol presented in red wine has potent cardiovascular effect in animal. Here, we investigated the ability of resveratrol to relax human coronary bypass grafts, saphenous vein and internal mammary artery and also its effect on their endothelial reactivity. METHODS: Vascular rings were obtained from 38 male patients undergoing coronary artery bypass operation. The relaxant effects of resveratrol (10-70 microM) and acetylcholine (10(-8)-10(-4) M) were examined on precontracted saphenous vein and internal mammary artery rings. RESULTS: Resveratrol, at concentration of 70 microM caused relaxations of 34.2+/-5.7% in saphenous vein and 35.2+/-5.4% in internal mammary artery. Endothelium removal and l-NOARG (nitric oxide synthase inhibitor, 10(-4) M) pretreatment almost completely inhibited the relaxation to resveratrol in internal mammary artery but partially in saphenous vein rings. Indomethacin (cyclooxygenase inhibitor, 10(-5) M) slightly, but not significantly enhanced the relaxation to resveratrol in both vessels. The endothelium-dependent relaxations to acetylcholine were significantly improved in the presence of resveratrol of 20 microM in both grafts (E(max): 33.8+/-3.7% versus 46.8+/-4% in saphenous vein n=9; p<0.05; 54. 4+/-5.3% versus 69.3+/-5.4% in internal mammary artery, n=8, p<0.05). The relaxations to acetylcholine were fully eliminated by combination of resveratrol with l-NOARG (10(-4) M) in both vessels. CONCLUSIONS: Resveratrol produced mainly endothelium-dependent and nitric oxide-mediated vasodilation in human internal mammary artery but partially in saphenous vein rings and improved their endothelial reactivity. This may have a therapeutic potential in cardiovascular diseases. PMID- 16243117 TI - Hyperkalemia induced failure of atrial and ventricular pacemaker capture. AB - Hyperkalemia is a life threatening metabolic condition. The common risk factors for hyperkalemia include renal insufficiency, use of angiotensin converting enzyme inhibitors, potassium supplementation and excessive consumption of potassium containing diet. A mild to moderate increase in serum potassium causes an increase in myocardial excitability, but further increase leads to impaired myocardial responsiveness, including that to pacing stimulation. Hyperkalemia has been reported to cause failure of atrial capture due to pacemaker exit block. We report a case where hyperkalemia resulted in failure of both the atrial and the ventricular pacemaker capture. PMID- 16243118 TI - Localized dissection of the descending thoracic aorta after blunt chest trauma. PMID- 16243119 TI - Constrictive pericarditis with persistent left superior vena cava flow into right atria through coronary sinus demonstrated by multislice computed tomography. PMID- 16243120 TI - Sub-pulmonary ventricular failure resulting from pulmonary hypertension in transposition of the great arteries palliated by atrial switch. PMID- 16243122 TI - Acute myocardial infarction in a young woman with antiphospholipid syndrome and occasional cocaine abuse. AB - We describe a case of acute myocardial infarction in a 19-year-old woman reporting a history of occasional cocaine abuse with last exposure 4 days before symptom onset, otherwise at low cardiovascular risk. Coronary angiography showed thrombotic occlusion of anterior descending coronary artery without atherosclerotic plaques, and complete recanalization of the vessel after anticoagulation with enoxaparine. Hypercoagulability evaluation revealed the presence of antiphospholipid antibodies. We suppose that cocaine abuse may have had a delayed effect as trigger of acute myocardial infarction. This action may be due to cocaine-induced endothelial activation and to a synergic prothrombotic activity of cocaine and antiphospholipid antibodies. PMID- 16243121 TI - Staphylococcus aureus infective endocarditis and septic pulmonary embolism after septic abortion. AB - We report a case of a 26-year-old female who after septic abortion developed tricuspid valve endocarditis with large vegetations, which subsequently resulted in embolism to right main pulmonary artery. Patient was treated with intravenous antibiotics and had a good recovery. PMID- 16243123 TI - Molecular diagnosis of infectious diseases in dermatology. AB - The molecular diagnosis of infectious disease has been growing considerably over the past decade. Nucleic acid amplification techniques, such as polymerase chain reaction, ligase chain reaction, transcription-mediated amplification, and nucleic acid sequence-based amplification, provide highly accurate diagnosis of numerous bacterial, viral, fungal, and parasitic infections involved in a variety of dermatologic diseases. In addition, signal amplification with hybrid capture, branched-DNA assays, and in situ hybridization have been used to detect numerous viral pathogens with high degrees of sensitivity and specificity. New technology that involves the use of DNA and protein microarrays has also enabled the detection of a variety of genes and gene mutations. With time, these diagnostic assays are decreasing in cost, gaining approval of the U.S. Food and Drug Administration, and becoming easier and more efficient to use. In the future, these assays will be able to deliver rapid and accurate diagnosis of infectious diseases within a single clinic visit. LEARNING OBJECTIVE: At the completion of this learning activity, participants should be familiar with molecular diagnosis of infectious diseases in dermatology. PMID- 16243124 TI - Long-term treatment of photoaged human skin with topical retinoic acid improves epidermal cell atypia and thickens the collagen band in papillary dermis. AB - BACKGROUND: Risk of photocarcinogenesis and the relevance of collagen in wrinkle effacement are two issues related to prolonged use of retinoic acid (RA) that have not been fully addressed. OBJECTIVE: Our purpose was to investigate the degree of epidermal cellular atypia and the thickness of papillary dermal collagen in photoaging after long-term use of RA. METHODS: Thirty-four subjects with photoaged skin were treated daily with 0.05% RA for at least 6 months. Epidermal cellular atypia was graded by means of a semiquantitative scale. Thickness of collagen band was measured by using image-analysis software. RESULTS: Compared with pretreatment findings, melanocytic and keratinocytic atypia was significantly reduced and the collagen band thickness doubled. LIMITATIONS: This was an open-label study. CONCLUSION: Improvement in epidermal cellular atypia is consistent with the ability of RA to act as a chemopreventive agent in epithelial carcinogenesis. Prolonged use also significantly increased collagen matrix deposition in dermal repair zones, which most likely contributes to wrinkle effacement by RA. PMID- 16243125 TI - Dermal matrix remodeling after nonablative laser therapy. AB - OBJECTIVE: Nonablative laser therapy is widely practiced for cutaneous rejuvenation. We sought to quantify dermal molecular changes after exposure of photodamaged skin to nonablative laser energy. METHODS: Nonablative laser therapy of forearm skin using either a 585-nm wavelength pulsed dye laser or a 1320-nm wavelength neodymium:yttrium-aluminum-garnet laser was performed. Serial biopsy specimens were obtained at baseline and various times after treatment. RESULTS: Statistically significant increases in type I procollagen messenger RNA expression occurred after exposure of photodamaged skin to each laser. Induction was 47% (P < .05) and 84% (P < .05) above baseline levels 1 week after laser therapy among those treated with the pulsed dye and neodymium:yttrium-aluminum garnet lasers, respectively. Substantial induction of type III procollagen, various matrix metalloproteinases, and primary cytokines was also demonstrated. Responses with respect to all molecules studied were highly variable. LIMITATIONS: This study addresses molecular changes after a single laser exposure whereas clinically, serial treatments are often provided. CONCLUSIONS: Nonablative laser therapy may result in quantifiable alterations in molecules associated with remodeling of the dermal matrix, although responses vary greatly among patients. PMID- 16243126 TI - Skin cancer in organ transplant recipients: effect of pretransplant end-organ disease. AB - BACKGROUND: Solid organ transplant recipients are at increased risk for posttransplant neoplasms. OBJECTIVE: Our purpose was to determine whether various diseases causing end-organ failure are associated with different degrees of risk of skin cancer development after transplantation. METHODS: The Organ Procurement and Transplantation Network/United Network for Organ Sharing Transplant Tumor Registry was searched for the incidence of skin cancer among kidney, liver, and heart transplant recipients in the United States between 1996 and 2001. Multivariate analysis was used to determine the association between disease diagnosis and posttransplant skin cancer. RESULTS: Transplant recipients with specific pretransplant diseases, such as polycystic kidney disease and cholestatic liver disease, were at increased risk for skin cancer. Patients with diabetes mellitus had a lower incidence of skin cancer after kidney transplantation. LIMITATIONS: The study had only a brief follow-up period, indirect assessment of photodamage, and possible underreporting. CONCLUSION: Transplant recipients with a history of certain diseases warrant intensive skin cancer surveillance and strict sun-protective practices. PMID- 16243127 TI - A randomized, double-blind, placebo-controlled trial of the combined effect of doxycycline hyclate 20-mg tablets and metronidazole 0.75% topical lotion in the treatment of rosacea. AB - BACKGROUND: Subantimicrobial doses of doxycycline may improve outcomes in rosacea when combined with topical metronidazole and used as maintenance monotherapy. OBJECTIVE: The purpose of this study was to evaluate the safety and efficacy of doxycycline hyclate 20 mg (subantimicrobial dose doxycycline) administered twice daily as an adjunct to metronidazole 0.75% topical lotion in the treatment of rosacea. METHODS: Patients received subantimicrobial doses of doxycycline twice daily plus metronidazole (n = 20) or placebo plus metronidazole (n = 20) for 12 weeks. Subantimicrobial-dose doxycycline or placebo monotherapy continued for 4 weeks. The primary efficacy measure was change from baseline in total inflammatory lesions at weeks 2 and 16. RESULTS: Total inflammatory lesions were reduced significantly (P =.048) by week 4 and by all subsequent visits in the subantimicrobial-dose doxycycline/metronidazole group compared with placebo/metronidazole. Changes from baseline increased over time and were maintained during subantimicrobial-dose doxycycline monotherapy. CONCLUSION: Adjunctive use of subantimicrobial dose doxycycline significantly reduced the clinical signs of rosacea compared with metronidazole alone and may be useful maintenance monotherapy. PMID- 16243128 TI - A randomized controlled trial assessing the efficacy of fluconazole in the treatment of pediatric tinea capitis. AB - BACKGROUND: Griseofulvin is considered first-line therapy for tinea capitis, and the Physician's Desk Reference currently recommends 11 mg/kg per day microsize formulation for use in children. Diverse selective pressures have resulted in waning clinical efficacy of griseofulvin, such that higher doses and longer courses of treatment are required. These events have prompted the search for therapeutic alternatives. Fluconazole is one such treatment option, and a variety of studies using this drug have shown promise in the treatment of pediatric tinea capitis. OBJECTIVE: We sought to assess the efficacy, safety, and optimal dose and duration of fluconazole therapy compared with standard-dose griseofulvin (11 mg/kg per day microsize formulation) in the treatment of pediatric tinea capitis. METHODS: This randomized, multicenter, third-party-blind, 3-arm trial was designed as a superiority study to identify a therapeutically superior agent/regimen from the 3 treatment arms: (1) fluconazole 6 mg/kg per day for 3 weeks followed by 3 weeks of placebo, (2) fluconazole 6 mg/kg per day for 6 weeks, and (3) griseofulvin 11 mg/kg per day for 6 weeks. Efficacy variables included mycological, clinical, and combined outcomes. The primary efficacy variable was the combined outcome of the modified intent-to-treat population at week 6. Patient safety was assessed throughout the study. Statistical analysis of the efficacy variables was conducted by means of the Cochran-Mantel-Haenszel test. RESULTS: At the end of treatment, mycological cures were present in 44.5%, 49.6%, and 52.2% of the fluconazole 3-week, fluconazole 6-week, and griseofulvin groups, respectively. Analysis of the primary efficacy variable failed to identify any superior agent, and differences between the combined outcomes of the fluconazole 6-week and griseofulvin groups at week 6 were not significant (P = .32). Regarding mycological, clinical, and combined outcomes, no significant differences between the fluconazole 6-week and griseofulvin groups were detected at any time point in the study. No new safety concerns were raised by this trial, and the incidence of treatment-related adverse events noted in this study is concordant with previous reports. Patients in the fluconazole arms of the study fared similarly. At the end of the trial, the difference in mycological cures between the fluconazole arms was only 7.5%, and increases in the incidence of certain treatment-related adverse events were observed in the fluconazole 6-week group. LIMITATIONS: Adjunctive topical therapies and the impact of infected contacts were not assessed in this trial. CONCLUSION: Systemic therapy with fluconazole 6 mg/kg per day and standard-dose griseofulvin produces comparable but low mycological and clinical cure rates. The limited efficacy of standard dose griseofulvin and the lack of consensus regarding dose and duration of griseofulvin therapy in tinea capitis emphasize the need for controlled trials to identify optimal treatment parameters. Although the efficacy of fluconazole is no better than that of standard-dose griseofulvin, it may still be useful in select patients with a contraindication or intolerance to high-dose griseofulvin. The outcomes observed in this trial highlight the need to more clearly define the relative importance of adjunctive topical therapies and the evaluation and treatment of infected contacts as factors affecting cure rates. PMID- 16243129 TI - Leishmania tropica in northern Israel: a clinical overview of an emerging focus. AB - BACKGROUND: In Israel, most cutaneous leishmaniasis (CL) is caused by Leishmania major. Recently a new focus of CL caused by Leishmania tropica has been described in Tiberias and the surrounding area of northern Israel. OBJECTIVE: The aim of this study was to evaluate clinical (size, number, location, and type of lesion) and laboratory (culture and polymerase chain reaction [PCR] analysis) parameters at diagnosis, response to treatment, and outcome of patients with CL due to L tropica. METHODS: Between September 2002 and March 2004, patients with direct smear-confirmed CL were evaluated; clinical records were reviewed and a telephone survey was performed. RESULTS: Forty nine patients, 34 (69%) male and 15 (31%) female, were studied. Mean age was 31.1 years (median 26 years, range 1-70); 76% of patients live in Tiberias and the surrounding area. The mean number of lesions was 2.6 (median 2, range 1-10). Lesions were commonly located on the face (61%) and upper limbs (57%). PCR analysis was performed in 27 patients and was positive for L tropica in 26. Fifty percent of patients studied received multiple therapeutic regimens because of incomplete response or treatment failure. Topical paromomycin was used in 44 patients (90%), with a complete response reported in only 17 (39%); of the 9 patients treated with intralesional sodium stibogluconate, a complete response was reported in 6 (67%); of the 5 patients treated with intravenous sodium stibogluconate, 4 (80%) were cured. LIMITATIONS: The relatively small number of patients studied combined with the fact that some were assessed retrospectively limit our conclusions. In addition, 50% of the patients studied received multiple therapeutic regimens because of failure of, or incomplete responses to, their initial therapy, thereby making comparisons difficult. CONCLUSIONS: The cure rate in those completing a course of antimony therapy, either 10 or more days of intravenous therapy or therapy administered intralesionally, was 75% (95% confidence interval [CI], 50.5-99.5%) as compared with 45% (95% CI, 28.9-60.5%) among those completing at least 10 days of topical paromomycin. To date, no standardized, simple, safe, and highly effective regimen for treating L tropica exists. Large, controlled clinical trials to evaluate current treatment regimens as well as new medications for CL, and especially CL attributed to L tropica, are urgently needed. PMID- 16243130 TI - Malignant melanoma in childhood and adolescence: report of 13 cases. AB - We reviewed all cases of malignant melanoma in children younger than 17 years of age who were evaluated at Sainte Justine Hospital, a tertiary care pediatric center, between 1980 and 2002. The medical records and histologic features of all cases were reviewed. Thirteen cases were identified, 4 boys and 9 girls. Fifty three percent of patients were prepubescent. None of the patients had a predisposing condition (eg, giant congenital nevi, dysplastic nevus syndrome, or xeroderma pigmentosum). One patient had had chemoradiotherapy previously for an undifferentiated pleuropulmonary malignant tumor (blastoma) and another patient had Down syndrome. The most frequent reason for initial consultation was a recent increase in size of the lesion. Three patients had pyogenic granuloma-like lesions. Eighty-five percent of the observed melanomas were nodular in type. Tumor thickness ranged from 0 to 6 mm with a median and mean thickness of 2.8 and 3.2 mm, respectively. The overall 5-year survival rate was 58.8%. Lack of awareness and delay in diagnosis may lead to a higher incidence of thick and intermediate melanoma in children. Because it appears that the majority of melanomas in childhood and adolescence occur de novo, clinicians should consider this condition in the differential diagnosis of any suspect lesion in children and adolescents even without an identified predisposing factor. PMID- 16243132 TI - Dermal tumescent local anesthesia in cutaneous surgery. PMID- 16243131 TI - Efficacy of 3 different light doses in the treatment of actinic keratosis with 5 aminolevulinic acid photodynamic therapy: a randomized, observer-blinded, intrapatient, comparison study. AB - BACKGROUND: Topical 5-aminolevulinic acid-based photodynamic therapy (ALA-PDT) has been established in recent years as an effective treatment for disseminated actinic keratosis (AK). As yet, however, data are lacking to define the optimal light dose for activation of ALA-induced protoporphyrin IX in AK. OBJECTIVE: In the present study our purpose was to compare the efficacy and tolerability of 3 different doses of red light for ALA-PDT of AK. METHODS: Twenty-seven patients with at least 3 clearly definable, mild or moderate AKs on the scalp or face entered the study. After occlusion for 4 hours with 20% ALA, one AK each was irradiated at random with a single dose of 70, 100, or 140 J/cm2. PDT-induced pain was assessed by the patients by means of a visual analog scale that graded pain intensity between 0 and 10. Follow-up examinations were performed 1 and 3 months after PDT. RESULTS: One month after PDT, the rate of complete remission (CR) was 89% for 70 J/cm2, 92% for 100 J/cm2, and 81% for 140 J/cm2. The CR rates at 3 months were 81% for 70 J/cm2, 77% for 100 J/cm2, and 69% for 140 J/cm2. No significant difference in therapeutic efficacy was found among the 3 light doses at either 1 month (P = .36) or 3 months (P = .96) after PDT. The degree of PDT induced pain during irradiation was substantial and not statistically different (P = .06) for all 3 light doses. LIMITATIONS: The conclusions from this study are limited by the small sample size and only apply to topical ALA-PDT. CONCLUSION: Our results indicate that a red light dose of 70 J/cm2 may be sufficient for effective topical ALA-PDT of disseminated, mild to moderate AK on the face and scalp. PMID- 16243133 TI - The 30-degree angle revisited. AB - The standard surgical ellipse, with 30 degrees apical angles and a length-to width ratio of 2 or 3 to 1, works optimally on a flat surface. The same pattern, when used for excisions on strongly convex or concave surfaces, leads to distortions which may require significant revisions. The reason for these discrepancies is explainable by the mathematical differences between flat Euclidian geometry and curved non-Euclidian geometry. Understanding these basic mathematical principles as applied to cutaneous surgery should lead to better preoperative planning, fewer intraoperative surprises, and more pleasing results. PMID- 16243134 TI - Basal cell carcinomas with unusual histologic patterns. AB - Uncommon histologic variants of basal cell carcinoma (BCC) can present a diagnostic challenge. In this case series, we describe 3 patients with unusual BCCs encountered in a dermatologic surgical unit over a 1-year period from September 2003 to September 2004. The formalin-fixed, paraffin-embedded histologic specimens were initially examined microscopically after staining with hematoxylin and eosin. Additional stains, including diastase periodic acid Schiff, colloidal iron, carcinoembryonic antigen, and cytokeratin-20, were subsequently performed as appropriate. Of the 3 lesions, one exhibited apocrine differentiation and two demonstrated a trabecular growth pattern. Although BCCs demonstrating apocrine differentiation have previously been described, a trabecular growth pattern, to our knowledge, has not been previously reported for BCC. PMID- 16243135 TI - Solitary cutaneous dendritic cell tumor in a child: role of dendritic cell markers for the diagnosis of skin Langerhans cell histiocytosis. AB - We describe a child with a solitary dendritic cell (DC) tumor positive for S-100 protein, CD1a, and HLA-DR with the clinical and histopathologic features of a so called solitary variant of congenital self-healing Hashimoto-Pritzker reticulohistiocytosis (CSHRH). CSHRH is a spontaneously regressing, benign form of Langerhans cell histiocytosis (LCH) and was thought to be a histiocytosis consisting of precursor Langerhans cells. In our study the tumor cells did not express CD68, indicating that they represent mature DCs. Because of the negative finding for Langerin, it cannot be assessed whether the tumor consists of terminally mature Langerhans cells that have lost Langerin expression upon maturation or of mature dermal DCs. This case demonstrates that the progress in DC biology necessitates reevaluation of our knowledge of LCH to better understand the different variants of the disease. Therefore the literature on CSHRH is reviewed in light of present knowledge on cutaneous DC immunology. PMID- 16243137 TI - After the tsunami. PMID- 16243136 TI - A systematic review of contact dermatitis treatment and prevention. AB - BACKGROUND: Contact dermatitis (CD) is a common occupational disease. There have been no systematic reviews of CD treatment or prevention. METHODS: Multiple databases were systematically searched. Using independent double review and published quality review criteria, articles were rated as good, fair, or poor. Treatment benefit data were tabulated and conclusions were based on the rated strength of published evidence. RESULTS: In all, 49 studies met inclusion criteria. Barrier creams containing dimethicone or perfluoropolyethers, cotton liners, and softened fabrics prevent irritant CD. Lipid-rich moisturizers both prevent and treat irritant CD. Topical skin protectant and quaternium 18 bentonite (organoclay) prevent rhus dermatitis. Diethylenetriamine pentaacetic acid (chelator) cream prevents nickel, chrome, and copper dermatitis. Potent or moderately potent steroids effectively treat allergic CD. There were no macrolide immunomodulator trials that met inclusion criteria. This review did not include studies of children, animals, or non-English language publications. CONCLUSIONS: A limited number of interventions effectively prevent or treat irritant and allergic CD, but well-controlled, outcome-blinded studies, particularly in the area of allergic CD prevention are needed. PMID- 16243138 TI - Trichorhinophalangeal syndrome. PMID- 16243139 TI - Antibiotic issues in dermatology. PMID- 16243140 TI - Medical analogies: their role in teaching dermatology to medical professionals and patients. PMID- 16243141 TI - Medical pearl: Using tissue transglutaminase antibodies to diagnose dermatitis herpetiformis. PMID- 16243142 TI - Medical pearl: Interpretation of tuberculin skin tests in patients who have received the BCG vaccine. PMID- 16243143 TI - Medical pearl: Blue underpants sign--a diagnostic clue for Pseudomonas aeruginosa intertrigo of the groin. PMID- 16243144 TI - Surgical pearl: Patient-applied manual pressure for hemostasis. PMID- 16243145 TI - Surgical pearl: The use of endarterectomy scissors in dermatologic surgery. PMID- 16243146 TI - Chronic edema of the lips--a rare but real problem: a report of 3 cases and their response to therapy. AB - Chronic edema of the lips is rare but disabling. We present 3 illustrative cases of chronic edema involving the lips. The causes were (1) idiopathic, (2) lymphedema occurring after radiation therapy, and (3) cheilitis granulomatosa. The edema subsided with compression therapy in the first 2 patients. Surgical decompression was helpful in the second patient, and dapsone therapy was initiated in the third. Chronic edema of the lips can be a clinically significant problem. These 3 cases illustrate the various causes and an approach to the management of chronic edema of the lips. PMID- 16243147 TI - Contact sensitization to Compositae mix in children. AB - The prevalence of contact sensitization to Compositae mix was investigated in 641 consecutive children. Seventeen children (12 with atopic dermatitis) tested positive for Compositae mix. We suggest adding Compositae mix to a pediatric screening series when investigating airborne dermatitis in children with atopic dermatitis. PMID- 16243148 TI - Symptomatic treatment of idiopathic and rosacea-associated cutaneous flushing with propranolol. AB - Flushing has been associated with medications, rosacea, menopause, carcinoid syndrome, pheochromocytoma, polycythemia, and mastocytosis, although it can occur without known cause. There are no known specific treatments available, but beta blockers have suppressed flushing reactions in some patients, particularly when associated with anxiety. The medical histories and clinical characteristics of 9 patients with either idiopathic flushing or flushing associated with rosacea were reviewed. Eight patients experienced subjective improvement with propranolol therapy. PMID- 16243149 TI - Dermoscopy of the nail bed and matrix to assess melanonychia striata. AB - Melanonychia striata represents a diagnostic dilemma for dermatologists. The use of dermoscopy to assess the nail has advantages over clinical examination. However, when compared to skin lesions, it gives fewer details. We describe two cases of melanonychia striata submitted to dermoscopic examination of the nail bed and matrix. This is a new procedure that enables observing dermoscopic characteristics that are not visualized in the nail plate, thus, providing additional information. PMID- 16243150 TI - Etanercept improves the health-related quality of life of patients with psoriasis: results of a phase III randomized clinical trial. AB - In this randomized, double-blind, phase III trial, patients with psoriasis received etanercept for 24 weeks or placebo for 12 weeks followed by etanercept for 12 weeks. At week 12, improvement in Dermatology Life Quality Index was 47% to 61% with etanercept compared with 11% with placebo (P < .0001). Etanercept rapidly and substantially improved patients' health-related quality of life. PMID- 16243151 TI - Frequent varicella zoster reactivation associated with therapeutic use of arsenic trioxide: portents of an old scourge. AB - In 44 patients treated with arsenic trioxide (As2(O3)) for acute promyelocytic leukemia, 11 developed varicella zoster virus (VZV) reactivation (median 56 days [range 15-299]) after treatment. There was no preferential dermatome involvement or systemic spread. The actuarial risk of VZV reactivation at 1 year was 26%. No VZV reactivation occurred after the first year of initial treatment with As2(O3). PMID- 16243152 TI - Sternal erythema: a distinctive postsurgical eruption. AB - BACKGROUND/OBJECTIVE: Wound complications constitute a diverse array of surgical and dermatologic entities. We sought to describe the clinical and pathologic attributes of a series of patients in whom a distinctive thoracic surgical wound eruption developed after coronary bypass grafting. METHODS: We performed a detailed chart review, including history, surgical procedure notes, and biopsy findings, obtained from 3 patients; we examined the data and reconciled them with conventional etiologies in an attempt to discern a shared diagnosis and pathogenesis. RESULTS: The patients were white men (59, 68, and 73 years of age) in whom roughly symmetrical asymptomatic erythema developed within the borders of their sternal thoracotomy wounds between 1 and 2 years after open heart surgery. The cutaneous findings consisted of macular erythema and poikiloderma that blanched with external pressure. Biopsy findings yielded epithelial atrophy with capillary telangiectases. CONCLUSION: Although cellulitis, dermal hypersensitivity to surgically implanted hardware including sternal wire, and an anatomic variant of costal fringe remain as possible diagnoses, an additional consideration includes a form of postsurgical reflex sympathetic dystrophy. PMID- 16243153 TI - CD8(+) T lymphocytes reactive against Epstein-Barr virus antigens in skin lesions of a patient with Sezary syndrome. AB - Several studies have investigated the possible involvement of viral agents, and among them herpes viruses, in the development of cutaneous T-cell lymphoma. The aim of our study was to determine whether T cells specific to Epstein-Barr virus (EBV) antigens were detectable among tumor-infiltrating lymphocytes infiltrating cutaneous lesions of a patient with Sezary syndrome. To analyze responses to EBV, we used a transient SV-40 origin-defective transformed simian cells transfection assay that permits an estimation of CD8 T-cell responses against a large number of HLA/viral protein combinations. This technique allowed the detection of EBV specific T lymphocytes mainly directed against epitopes generated during the lytic cycle in the cutaneous lesions. This is, to our knowledge, the first description of the presence of EBV-specific T lymphocytes among tumor infiltrating lymphocytes infiltrating the lesional skin of a patient with Sezary syndrome. PMID- 16243154 TI - Paintball purpura. PMID- 16243155 TI - The PUVA-turban as an alternative treatment of alopecia associated with chronic graft versus host disease. PMID- 16243156 TI - Infliximab treatment failure in a case of cutaneous Crohn's disease. PMID- 16243157 TI - Cutis marmorata telangiectatica congenita in unique association with imperforate anus and myelodysplasia. PMID- 16243158 TI - Persistent microscopic primary melanoma in patients presenting with node-positive disease. PMID- 16243159 TI - Kava-induced urticaria. PMID- 16243160 TI - Squamous cell carcinoma arising from lichen planus of nail matrix and nail bed. PMID- 16243161 TI - Toxic epidermal necrolysis secondary to timolol, dorzolamide, and latanoprost eyedrops. PMID- 16243162 TI - Topical 5-aminolevulinic acid-based photodynamic therapy for the treatment of a patient with cutaneous pseudolymphoma. PMID- 16243163 TI - Characterization of Epstein-Barr virus-infected natural killer lymphocytes in a patient with hypersensitivity to mosquito bites. PMID- 16243164 TI - Multiple mucinous (myxoid) cysts of a single finger. PMID- 16243165 TI - Severe chronic urticaria: response to mirtazapine. PMID- 16243166 TI - Ulcerative cutaneous sarcoidosis responding to adalimumab. PMID- 16243167 TI - Epidemiologic aspects of rosacea. PMID- 16243168 TI - Sun protection factor content and warning statements for sunless tanning products: an examination of retail outlets and the Internet. PMID- 16243170 TI - Are life insurance underwriting practices fair? PMID- 16243169 TI - A win-win proposition: fostering US health care consumer involvement in the Cochrane Collaboration Skin Group. PMID- 16243171 TI - Readiness and response: the oral and maxillofacial surgeon's role in disaster. PMID- 16243172 TI - Bisphosphonate-induced exposed bone (osteonecrosis/osteopetrosis) of the jaws: risk factors, recognition, prevention, and treatment. AB - PURPOSE: Bisphosphonates inhibit bone resorption and thus bone renewal by suppressing the recruitment and activity of osteoclasts thus shortening their life span. Recently three bisphosphonates, Pamidronate (Aredia; Novartis Pharmaceuticals, East Haven, NJ), Zoledronate (Zometa; Novartis Pharmaceuticals), and Alendronate (Fosamax; Merck Co, West Point, VA) have been linked to painful refractory bone exposures in the jaws. MATERIALS AND METHODS: One hundred nineteen total cases of bisphosphonate-related bone exposure were reviewed. RESULTS: Thirty-two of 119 patients (26%) received Aredia, 48 (40.3%) received Zometa, 36 (30.2%) received Aredia later changed to Zometa, and 3 (2.5%) received Fosamax. The mean induction time for clinical bone exposure and symptoms was 14.3 months for those who received Aredia, 12.1 months for those who received both, 9.4 months for those who received Zometa, and 3 years for those who received Fosamax. Sixty-two (52.1%) were treated for multiple myeloma, 50 (42%) for metastatic breast cancer, 4 (3.4%) for metastatic prostate cancer and 3 (2.5%) for osteoporosis. Presenting findings in addition to exposed bone were 37 (31.1%) asymptomatic, 82 (68.9%) with pain, 28 (23.5%) mobile teeth, and 21 (17.6%) with nonhealing fistulas. Eighty-one (68.1%) bone exposures occurred in the mandible alone, 33 (27.7%) in the maxilla, and 5 (4.2%) occurred in both jaws. Medical comorbidities included the malignancy itself 97.5%, previous and/or maintenance chemotherapy 97.5%, Dexamethasone 59.7%. Dental comorbidities included the presence of periodontitis 84%, dental caries 28.6%, abscessed teeth 13.4% root canal treatments 10.9%, and the presence of mandibular tori 9.2%. The precipitating event that produced the bone exposures were spontaneous 25.2%, tooth removals 37.8%, advanced periodontitis 28.6%, periodontal surgery 11.2%, dental implants 3.4% and root canal surgery 0.8%. CONCLUSIONS: Complete prevention of this complication in not currently possible. However, pre-therapy dental care reduces this incidence, and non-surgical dental procedures can prevent new cases. For those who present with painful exposed bone, effective control to a pain free state without resolution of the exposed bone is 90.1% effective using a regimen of antibiotics along with 0.12% chlorohexidine antiseptic mouth. PMID- 16243173 TI - The role of facial trauma as a possible etiologic factor in temporomandibular joint disorder. AB - PURPOSE: Facial trauma has been suggested as a possible etiologic factor of temporomandibular joint disorder. However, there is little information on the role of macrotrauma. The main purpose of this study was to validate facial trauma as a potential etiologic factor for temporomandibular joint (TMJ) disorder. Multidirectional approaches were applied for the evaluation of the changes of TMJ after TMJ macrotrauma. PATIENTS AND METHODS: Analysis of TMJ status including arthroscopic examination, histomorphologic examination, and synovial fluid biochemical analysis were performed on the patients with mandibular fractures. Additionally, the efficacy of arthrocentesis for the patients of mandibular fracture was evaluated from the functional point of view. RESULTS: In arthroscopic examinations, evidence of synovitis with variable degrees was found. The representative findings are fibrillation and ecchymosis. On histomorphologic examination, bloody smear, degenerated cells and cartilage, inflammatory cells, and crystal were observed. In biochemical analysis, considerable amounts of prostaglandin E(2) and leukotriene B(4) were detected in the synovial fluid of the patients. CONCLUSION: The inflammatory and degenerative changes of TMJ can develop after facial trauma. Trauma can be a possible etiologic factor in cartilage degeneration, and biochemical and intra-articular pathology. Clinicians should recognize the etiologic importance of macrotrauma, and long-term evaluation of the TMJ as well as adequate treatment is required for patients with facial trauma. PMID- 16243174 TI - Bilateral sagittal split osteotomy for correction of mandibular prognathism: long term results. AB - PURPOSE: To identify the long-term maxillomandibular changes after surgical correction of mandibular prognathism using bilateral sagittal split osteotomy (BSSO). PATIENTS AND METHODS: Twenty patients who underwent BSSO to setback the mandible and had cephalometric radiographs taken preoperatively and postoperatively at 6 weeks, 1 year, and long-term follow-up (mean, 28 months). The cephalograms were traced and measured to determine the operative and postoperative changes. Correlation analyses were performed to see the relationship between the magnitude of setback and the amount of long-term postsurgical change at B point and pogonion. RESULTS: The mean surgical setback was 8.2 mm at B point and 8.8 mm at pogonion. The mean long-term horizontal relapse was 2.3 mm (28.0%) at B point and 3.0 mm (34.1%) at pogonion. Out of 20 patients, 12 (60.0%) relapsed horizontally greater than 2 mm at B point and 13 (65.0%) at pogonion. The mean vertical surgical changes showed downward displacement of B point (2.3 mm) and pogonion (2.0 mm). The mean long-term vertical relapse was 1.6 mm (69.6%) at B point and 1.7 mm (85.0%) at pogonion. CONCLUSION: There was no correlation between the magnitude of setback and the amount of relapse at B point and pogonion. However, there was significant correlation between the magnitude of vertical, downward surgical displacement and the amount of vertical relapse at B point and pogonion. The majority of the maxillofacial changes occurred within 1 year postoperatively. PMID- 16243175 TI - Microbial analysis of the autogenous bone collected by bone filter during oral surgery: a clinical study. AB - PURPOSE: The purposes of the study presented are to identify the microbial content of the collected bone debris and to determine the antibacterial efficiency of chlorhexidine mouth rinse in reducing the microbial content of the collected bone debris. PATIENTS AND METHODS: Twenty-five patients who had asymptomatic fully impacted mandibular third molars bilaterally underwent surgical removal for prophylactic and orthodontic considerations. Immediately before surgery all patients rinsed with 10 mL of their assigned solution (chlorhexidine or sterile saline) for 2 minutes. Before surgical removal of the teeth, bone debris was collected with a stringent aspiration protocol from the ramus by bone filter. The microbial content of the bone debris was assessed and the bacterial levels between the 2 groups were compared statistically. RESULTS: All samples from both groups (chlorhexidine or sterile saline) yielded viable microorganisms. There was no significant difference between the mean/median colony-forming units (CFU)/g values of both group samples, for aerobes (Streptococcus salivarius) and anaerobes (Bacteroides, Peptococcus, Peptostreptococcus, and Veillonella species). With regard to total microorganisms, the mean CFU/g derived from the chlorhexidine group samples were 1.5 x 10(8) CFU/g per bone sample compared with 1.5 x 10(9) CFU/g for the sterile saline control group (P < .05). CONCLUSION: Although chlorhexidine rinsing immediately before surgery reduced the levels of total microorganisms when compared with stringent aspiration protocol alone, it has not been found effective on aerobic Streptococcus salivarius and, importantly, on anaerobes. The reduced bacterial levels may still carry high infectious risk and may lead to failure in autogenous grafting procedures in oral surgery. PMID- 16243176 TI - Prognostic factors in intraoral squamous cell carcinoma: the influence of histologic grade. AB - PURPOSE: The purpose of this retrospective study was to review the outcome of patients with oral cavity squamous cell carcinoma treated at a single institution by primary surgical resection with or without adjuvant radiotherapy or chemoradiotherapy and to identify factors affecting survival and locoregional control. PATIENTS AND METHODS: The records of 233 patients with oral cavity squamous cell carcinoma treated at a single institution from 1993 to 2003 were identified from the Legacy Emanuel Hospital and Health Center's cancer registry (Portland, OR). All patients undergoing surgical resection as a primary treatment modality were included in the study. Patients with nonresectable disease, distant metastasis, and those with inadequate follow-up data were excluded from the study. Patients with positive surgical margins, high-grade histology, aggressive biologic behavior, or advanced staged disease underwent adjuvant radiotherapy or chemoradiotherapy. The data collected included age, gender, race, tumor site, margin status, grade, TNM stage, cancer therapies, and cancer status. Data were statistically analyzed in an attempt to identify predictors of locoregional control and disease-free survival. Descriptive statistics were calculated for each variable and survival was calculated using the Kaplan-Meier method. Prognostic factors were analyzed using the Cox proportional hazard model. RESULTS: Two hundred fifteen patients consisting of 119 men (55%) and 123 females (52%), with an average age at diagnosis of 66 years (SD +/- 14), met the criteria for inclusion in the study. Average tumor size was 23.5 mm (SD +/- 14.1). Overall 5-year survival was 56% and disease-free survival at 5 years was 58%. Stage and grade were identified as having a statistically significant effect on survival (P = .014; likelihood ratio chi-square = 10.7, 3 degrees of freedom; and P = .026; likelihood ratio chi-square = 5, 1 degree of freedom, respectively). Neither age, gender, race, tumor site, nor positive margins showed a statistically significant effect on survival (P > .05). CONCLUSION: This study highlights the importance of grade and stage as independent factors in predicting survival in patients with oral squamous cell carcinoma. PMID- 16243177 TI - The assessment of quality of life in patients who have undergone surgery for oral cancer: a preliminary report. AB - PURPOSE: The outcome of management strategies for the treatment of oral cancer has always been in terms of disease-specific survival. Modern assessment of outcome now also includes assessment of quality of life (QOL). Little has been done previously in Australia to assess the QOL of patients treated primarily by surgery for oral cancer. MATERIALS AND METHODS: Twenty-two patients who had undergone primary surgical management for oral cancer underwent assessment of QOL using the European Organization for Research and Treatment of Cancer QOL questionnaire version 3 (EORTC QLQ-C30) and the University of Washington Head and Neck QOL questionnaire version 4 (UWQOL). Similar emotional domains were compared and factors affecting QOL were determined. RESULTS: Fifty-five percent of patients were male and 45% were female. The mean age was 61.8 years (range, 38 to 86 years) with a mean review time of 24.6 months (SD, 20.3 months) postoperatively. Good QOL scores were experienced by most patients. A new addition to the UWQOL version 4 was the mood and anxiety domains, and these correlated well with the emotional domains of the EORTC QLQ-C30 using Spearman rank correlations. Factors significantly affecting survival included age, overall stage, neck dissection, free flap, and postoperative radiotherapy using Mann Whitney tests (P < .05). No significant difference in QOL was found in patients reviewed at either less than or greater than 1 year. CONCLUSIONS: This study confirms previous reports in the literature. It has provided us with the foundation for further investigation of QOL in patients who have been and who will be treated for oral cancer. It also gives us a new measure of outcome to compare against survival rates. PMID- 16243178 TI - Cutaneous cysts of the head and neck. AB - PURPOSE: This article reviews the types of cutaneous cysts in patients referred to the Facial Lesion Clinic at John Peter Smith Hospital in Fort Worth, TX, and proposes effective treatment modalities based on lesion and patient variables. Cyst variables included proper identification, size of the lesion, and acute or chronic processes. Patient considerations included age, skin type, and location. Medical and social histories were not noted. PATIENTS AND METHODS: Eighty-two patients who had 1 or more cysts removed over the 5-year period from July 15, 1998 to July 14, 2003 were reviewed for age, gender, histologic diagnosis, anatomic location of the lesion, and complications. RESULTS: Patients with epidermal inclusion cysts (79%), followed by pilar cysts (9%), hidrocystomas and dermoid cysts (3% each), and multiple other diagnoses (less than 2%) were treated. Neither complications nor recurrent infections were reported during the 5-year interval. There were no recurrent cyst formations noted by return appointment. CONCLUSION: Cystic lesions of the head and neck may be treated effectively as long as they are correctly identified and treated in a specific manner. PMID- 16243179 TI - An in vitro comparison of an adjustable bone fixation system. AB - PURPOSE: The purpose of this study was to biomechanically compare an adjustable plating system and several variations with an inverted-L pattern of 3 bicortical screws to fix a bilateral sagittal split osteotomy. MATERIALS AND METHODS: Sixty polyurethane mandible replicas (Synbone, Laudquart, Switzerland) were used in this study. Ten uncut mandibular replicas served as control models. Fifty experimental synthetic mandibles had bilateral sagittal split osteotomies created with a 7-mm advancement. Fixation modalities included 3 bicortical screws in an inverted-L pattern, a 4-hole sagittal split plate with adjustable slider (slide 0), an adjustable plate with an additional bicortical screw (slide 1), an adjustable plate with 2 additional bicortical screws (slide 2), and an adjustable plate with 3 additional bicortical screws (slide 3). The alloplastic mandibles were secured in a custom fabricated jig and subjected to vertical loads at the incisal edge and torsional loading at the molar region by an Instron 1331 (Instron, Canton, MA) servohydraulic mechanical testing unit. Mechanical deformation data within a 0- to 900-N range were recorded. Yield load, yield displacement, and stiffness were determined. Means and standard deviations were derived and compared for statistical significance using a Fisher's Protected Least Significant Differences Test with a confidence level of 95% (P < .05). Second-order best-fit polynomials were created for the experimental data curves. RESULTS: For incisal edge loading, the control was significantly greater for yield load than the experimental models. No significant difference was noted between slide 0, slide 1, and slide 2, but significant differences were seen with slide 3 and the inverted-L. There were no significant differences between slide 1, slide 2, slide 3, and inverted-L. For molar load, the control was significantly different than the experimental models. However, there was no significant difference between the experimental models. CONCLUSIONS: Mandibular advancements of 7 mm in a synthetic mandible fixed with a 4-hole sagittal split plate with adjustable slider alone is resistant to torsional forces and comparable to that seen with 3 bicortical screws placed in an inverted-L fashion. However, when loading at the incisal edge, additional bicortical screws are necessary to achieve the similar resistance to vertical load as that seen with 3 bicortical screws. PMID- 16243180 TI - Bone regeneration in osseous defects using a resorbable nanoparticular hydroxyapatite. AB - PURPOSE: This animal study examined the de novo bone formation in bony defects following the insertion of autogenous bone alone versus an injectable nanoparticle hydroxyapatite alone and in combination with 25% autogenous bone. The regenerative potentials of the tested materials were compared with each other. MATERIALS AND METHODS: A model with biological similarity to humans with regard to bone regeneration was a prerequisite for the transferability of the results to clinical practice. Therefore, the adult domestic pig was the animal of choice. A total observation period of 6 months was selected. Microradiographic and histologic evaluation of the bone specimens was completed at 8 defined times. RESULTS: Microradiography indicated mineralization rates in the 2 bone substitute groups that were not significantly lower than those found in the autogenous bone group. Histologically, there was suitable osseointegration and osteoconduction of the used material. Complete resorption of the nanoparticle hydroxyapatite had taken place after 12 weeks. CONCLUSIONS: It can be concluded that the evaluated nanoparticular hydroxyapatite met the clinical requirements for a bone substitute material within the limits of this experimental setting. Due to its microstructure, complete resorption took place during the course of this study. PMID- 16243181 TI - Treatment options to optimize display of anterior dental esthetics in the patient with the aged lip. AB - PURPOSE: With aging a number of predictable changes occur in the upper and lower lips that act to mask otherwise esthetic dental procedures including dental implants, gingival esthetic surgery, and optimal dental restorations. The purpose of this article is to show how lip modification can be used to enhance the esthetic appearance of the patient, with specific attention to esthetic dental smile line characteristics. METHOD: Proper evaluation and diagnosis of senile lip changes, such as lip atrophy, loss of lip architecture, and lip lengthening lead to a reliable treatment plan that provides a more esthetic frame to view the oral aperture. Treatment options will be discussed that act to reverse these problems. Directed lip augmentation procedures are used to correct loss of lip volume and architecture. The subnasal lip lift technique is presented as a method to correct lip lengthening. Risks and limitations of these procedures are addressed. CONCLUSION: Safe and reliable methods are presented that can improve and rejuvenate the lip complex, thus allowing for optimal display of dental esthetics. PMID- 16243182 TI - Indications for splinting implant restorations. AB - PURPOSE: The purpose of this article was to review the literature concerning the need to splint implants together when restoring them with a provisional restoration immediately after implant placement. METHODS: The literature is reviewed concerning the rationale for splinting teeth and reports concerning the efficacy of splinting implants together. Based on this team's experience with a prospective series of consecutive 2 to 5 unit provisionalization cases, guidelines are included with case examples for understanding the technique. CONCLUSION: As long as canine guidance is present, and occlusion is stable, multiunit single quadrant restorations do not need to be splinted when provisionalized. PMID- 16243183 TI - Implant restoration of external resorption teeth in the esthetic zone. AB - PURPOSE: The purposes of this article were to review the literature for treatment of teeth with external resorption secondary to avulsive injuries and to illustrate treatment of patients with teeth following different clinical scenarios to develop a predictable course of therapy. MATERIALS AND METHODS: Cases that demonstrate treatment of teeth with external resorption following injury during the growing period, external resorption treated with a delayed approach after tooth extraction, immediate grafting of the extraction site to repair labial bone loss, and immediate implant placement with provisionalization are shown. CONCLUSION: Based on a review of the literature, the decision to place dental implants to replace teeth with external resorption can be timed depending on the location and type of the resorption, with excellent esthetic results. PMID- 16243184 TI - The treatment of odontogenic keratocysts by excision of the overlying, attached mucosa, enucleation, and treatment of the bony defect with carnoy solution. PMID- 16243185 TI - Treatment of keratocysts: the case for decompression and marsupialization. PMID- 16243186 TI - Intraorbital epidermoid tumors: a case report and a review of the literature. PMID- 16243187 TI - Unique pericoronal orocutaneous fistula: sequelae of chronic subclinical inflammation. PMID- 16243188 TI - Maxillomalar monoblock removal, reshaping, and reinsertion in Paget's disease: 15 year follow-up. PMID- 16243189 TI - Fixate: incorrect grammar. PMID- 16243190 TI - On alveolar distraction in the upper jaw. PMID- 16243191 TI - Biochemical and etiological characteristics of acute hyponatremia in the emergency department. AB - Hyponatremia can be classified as acute or chronic depending on its duration, and treatment options are tailored to this classification. However, it is sometimes difficult to differentiate acute from chronic hyponatremia in the Emergency Department (ED). The objective of this study was to identify characteristics to help diagnose and manage acute hyponatremia in the ED. Patients with acute hyponatremia in the ED were enrolled from a retrospective 2-year chart review. Eleven patients (0.8%) were identified with acute hyponatremia out of a total of 1321 hyponatremic patients. There were nine women and two men. The mean age was 48.9 years. The mean sodium (Na(+)) level was 115+/-4 mmol/L. Accompanying biochemical abnormalities included hypouricemia and hypouremia with increased fractional excretions of uric acid (UA) and urea. The estimated amount of water intake ranged from 2.5 to 10 liters (mean, 5.1+/-2.3 liters) during the day before ED presentation. All patients were treated with hypertonic saline and furosemide at a correction rate of 1.6+/-0.5 mmol/L/h. No patients had neurological sequelae after treatment. The causes of acute hyponatremia included induction of abortion with oxytocin (n=1), primary polydipsia on neuroleptic agents (n=2), polyethylene glycol (PEG) preparation for colonoscopy (n=1), diuretic therapy for hypertension (n=4), ecstasy use (n=1), and weight-reducing herbal teas (n=2). We conclude that in the right clinical setting, high free water intake and low serum urea and UA favor acute hyponatremia. A detailed drug history may be helpful in the differential diagnosis of acute hyponatremia. PMID- 16243192 TI - Severe imported malaria: clinical presentation at the time of hospital admission and outcome in 42 cases diagnosed from 1996 to 2002. AB - The objectives of this retrospective study were to describe initial clinical profiles and subsequent outcome of adult patients in France who were diagnosed with severe imported malaria, as defined by the World Health Organization (WHO). Forty-two patients diagnosed from 1996 to 2002 were included (median age: 30 years, men: 78%, non-immune persons: 74%, return from Africa: 100%, inappropriate antimalarial chemoprophylaxis: 95%). At the time of hospital admission, jaundice (62%), hyperparasitemia (56%), and prostration (52%) were the most frequent findings, followed by acute renal failure (31%). Other findings, as described by the WHO criteria, were less common. Twenty-three patients presented only with jaundice, hyperparasitemia, or prostration in isolation, or in combination. Of these 23, five non-immune persons subsequently developed coma, shock, acute respiratory distress syndrome or acute renal failure; this led to death in 2 of these cases. This suggests that non-immune persons with imported malaria who present with jaundice, hyperparasitemia, or prostration should be admitted to the intensive care unit for close monitoring. PMID- 16243193 TI - Prevalence of acute myocardial infarction and other serious diagnoses in patients presenting to an urban emergency department with chest pain. AB - A retrospective cohort study and chart review were performed to estimate the absolute and relative prevalence of the serious diagnoses that might cause a patient to present to the Emergency Department (ED) with a chief complaint of chest pain. In this study, we queried a database of 347,229 complete visits to the San Francisco General Hospital Emergency Department between July 1, 1993 and June 30, 1998 for visits by patients > 35 years old with a chief complaint of chest pain and no history of trauma. Visits for chest pain that resulted in hospitalization were assigned to one of nine diagnostic groups according to final diagnoses as coded in the database. Manual chart review by trained abstractors using explicit criteria was done when group assignment based on coded diagnoses was unclear and in all diagnoses of pulmonary embolism and aortic dissection. Of 8,711 visits (2.5% of all visits) with a chief complaint of non-traumatic chest pain, 3,271 (37.6%) resulted in hospitalization. Of the 3,078 (94.1% of those hospitalized) assigned a final diagnosis, 329 (10.7% of hospitalizations, 3.8% of all visits) had acute myocardial infarction, 693 (22.5%) had either unstable angina or stable coronary artery disease, and 345 (11.2%) had pulmonary causes (mainly bacterial pneumonia) deemed serious enough to require hospitalization. Pulmonary embolism and aortic dissection were diagnosed in only 12 (0.4%) and 8 (0.3%) patients, respectively. In 905 (29.4%) hospitalizations for chest pain, myocardial infarction was "ruled out" and no cardiac ischemia or other serious etiology for the chest pain was diagnosed. Among patients presenting with chest pain, those in older age groups had dramatically increased risk of acute myocardial infarction. Women presenting with chest pain had a lower risk of acute myocardial infarction than men. In conclusion, the prevalence of acute myocardial infarction in the undifferentiated ED patient with a chief complaint of chest pain is only about 4%. An equal number of patients will have a serious pulmonary cause as the etiology of their pain. Pulmonary embolism and aortic dissection are important but extremely rare causes of a chest pain presentation to the ED. PMID- 16243194 TI - The association between field Glasgow Coma Scale score and outcome in patients undergoing paramedic rapid sequence intubation. AB - Early intubation is standard for treating severe traumatic brain injury (TBI). Aeromedical crews and select paramedic agencies use rapid sequence intubation (RSI) to facilitate intubation after TBI, with Glasgow Coma Scale (GCS) score commonly used as a screening tool. To explore the association between paramedic GCS and outcome in patients with TBI undergoing prehospital RSI, paramedics prospectively enrolled adult major trauma victims with GCS 3-8 and clinical suspicion for head trauma to undergo succinylcholine-assisted intubation as part of the San Diego Paramedic RSI Trial. The following data were abstracted from paramedic debriefing interviews and the county trauma registry: demographics, mechanism, vital signs including GCS score, clinical evidence of aspiration before RSI, arrival laboratory values, hospital course, and outcome. Paramedic GCS calculations were confirmed during debriefing interviews. Patients were stratified by GCS score, with chi-square and receiver-operator-curve (ROC) analysis used to explore the relationship between GCS and hypoxia, head injury severity, aspiration, intensive care unit (ICU) length of stay, and outcome. Cohort analysis was used to explore potential reasons for early extubation and discharge from the ICU in some patients. A total of 412 patients were included in this analysis. A total of 81 patients (20%) were extubated and discharged from the ICU in 48 h or less; these patients had higher pre-RSI oxygen saturation (SaO(2)) values and higher arrival serum ethanol levels. Paramedic and physician GCS calculations had high agreement (kappa=0.995). A statistically significant relationship was observed between GCS score and Head Abbreviated Injury Score (AIS), survival, and pre-RSI SaO(2) values. However, ROC analysis revealed a limited ability of GCS to predict the presence of severe TBI, injury severity, desaturation, aspiration, ICU length of stay, or ultimate survival. In conclusion, paramedics seem to accurately calculate GCS values before prehospital RSI. Although a relationship between paramedic GCS and outcome exists, the ability to predict the severity of injury, airway-related complications, ICU length of stay, and overall survival is limited using this single variable. Other factors should be considered to screen TBI patients for prehospital RSI. PMID- 16243195 TI - Use of spiral computed tomography contrast angiography and ultrasonography to exclude the diagnosis of pulmonary embolism in the emergency department. AB - Spiral computed tomography (CT) contrast angiography is a promising imaging modality for the diagnosis of pulmonary embolism but the negative predictive value of this test remains controversial. We performed a multi-center prospective cohort study to determine the safety of relying on a negative spiral CT contrast angiography scan to exclude pulmonary embolism. Patients presenting to the Emergency Departments of three tertiary care institutions with clinically suspected pulmonary embolism were potentially eligible for the study. Patients underwent a clinical evaluation to categorize pretest probability into low, moderate, and high categories, and had D-dimer testing performed. Patients at low pretest probability with normal D-dimer were considered to have pulmonary embolism excluded. The remaining patients underwent spiral CT contrast angiography scan of the pulmonary arterial circulation and bilateral venous ultrasound of the proximal leg veins. Patients who were confirmed to have pulmonary embolism or deep vein thrombosis were treated with anticoagulant therapy. Patients in whom the diagnosis of pulmonary embolism was excluded did not receive anticoagulant therapy and were followed for a 3-month period for the development of venous thromboembolic complications. Eight hundred fifty-eight (858) patients were enrolled in this study. Three-hundred sixty-nine (369) patients had low pretest probability and negative D-dimer results and no further diagnostic tests were performed. None of these patients subsequently developed venous thromboembolic complications (0%, 95% confidence interval [CI] 0% to 1.0%). The remaining 489 were referred for spiral CT contrast angiography scan and ultrasound. Sixty-seven patients were confirmed to have pulmonary embolism and an additional 15 patients with negative CT scans had proximal deep vein thrombosis (DVT) on ultrasound for a total prevalence of venous thromboembolism of 82/489 (16.8%). Two of 409 patients who had pulmonary embolism excluded in the initial evaluation phase developed proximal venous thromboembolism (0.5%; 95% CI 0% to 1.8%) in the 3-month follow-up period. These findings suggest that the combination of a negative spiral CT contrast angiography scan and normal venous ultrasound imaging safely excludes the diagnosis of pulmonary embolism in the Emergency Department setting. PMID- 16243196 TI - Tissue plasminogen activator for acute ischemic stroke: a New York city emergency medicine perspective. AB - Nationally, only 2-3% of patients with acute ischemic stroke (AIS) currently receive tissue plasminogen activator (TPA). To better understand the reasons, we investigated the practice patterns, level of familiarity and acceptance of TPA for AIS among emergency physicians in New York City (NYC). Fifty-seven 911 receiving hospital emergency department directors were surveyed regarding TPA use. Of those responding, 37% had never used TPA to treat AIS. Lack of neurological support was reported by 33%. Departments with formal protocols were more likely to use TPA for AIS. In conclusion, there is considerable variation in the practice, knowledge, and attitudes regarding the use of TPA for AIS in NYC emergency departments. Improved educational efforts and institutional support may be necessary to ensure the appropriate use of TPA by emergency physicians. PMID- 16243198 TI - Does public education reduce ice storm-related carbon monoxide exposure? AB - Public education to prevent carbon monoxide exposure during ice storms has been recommended; its effects remain unexamined. We compared patients seen for carbon monoxide inhalation at the area's only academic Emergency Department during 1991 and 2003 ice storms; educational efforts were more intense in 2003. There were fewer patients during the second storm (45 vs. 55); all recovered fully. The percentage of Caucasian patients rose (from 57% to 89%) whereas that of African American patients fell (from 39% to 7%). Indoor grill use, associated with 11% of 1991 cases, was eliminated in 2003. Indoor gas generators remain the most common source. Carboxyhemoglobin levels correlate poorly with ambient carbon monoxide levels. Enhanced public education had a modest effect, especially in reducing the proportion of African-American patients and those from indoor grill use. Research on more effective public health education targeted at gas generator users and combined with physical interventions should be considered. PMID- 16243197 TI - The sensitivity of cardiac markers stratified by symptom duration. AB - We compared the sensitivity of three commonly used cardiac markers between two subpopulations, those who came to the Emergency Department (ED) late (6-24 h) after their symptoms began, and those who arrived earlier (<6 h), in a prospective comparative trial. Among all adult patients who presented to our ED with symptoms suggestive of acute myocardial infarction (MI), we drew serum for myoglobin, CK-MB, and troponin I upon arrival (time 0) and 2 h later. Outcomes, including acute MI, were determined. Sensitivities for all three markers between the subpopulations who arrived fewer than 6 h from symptom onset were compared to those who arrived later (6-24 h). We enrolled 346 eligible subjects, 36% of whom described cardiac symptoms as beginning 6 or more hours earlier; 14% suffered acute MIs. For time 0, the sensitivity of all three markers for acute MI was significantly higher among those subjects with symptoms of 6 or more hours' duration as compared to those with less. For troponin I, the increase in sensitivity between these two subpopulations approached 300%. At the time of the 2-h sample, the differences in sensitivities were much less and were not statistically significant. We conclude that cardiac marker values obtained at time 0 among Emergency Department patients who arrive 6 or more hours after cardiac symptom onset provide significantly higher sensitivities as compared to those obtained in patients who arrive earlier. For troponin I, the increase in sensitivity approaches threefold. PMID- 16243199 TI - Delayed presentation of traumatic facial nerve (CN VII) paralysis. AB - Facial nerve paralysis (Cranial Nerve VII, CN VII) can be a disfiguring disorder with profound impact upon the patient. The etiology of facial nerve paralysis may be congenital, iatrogenic, or result from neoplasm, infection, trauma, or toxic exposure. In the emergency department, the most common cause of unilateral facial paralysis is Bell's palsy, also known as idiopathic facial paralysis (IFP). We report a case of delayed presentation of unilateral facial nerve paralysis 3 days after sustaining a traumatic head injury. Re-evaluation and imaging of this patient revealed a full facial paralysis and temporal bone fracture extending into the facial canal. Because cranial nerve injuries occur in approximately 5 10% of head-injured patients, a good history and physical examination is important to differentiate IFP from another etiology. Newer generation high resolution computed tomography (CT) scans are commonly demonstrating these fractures. An understanding of this complication, appropriate patient follow-up, and early involvement of the Otolaryngologist is important in management of these patients. The mechanism as well as the timing of facial nerve paralysis will determine the proper evaluation, consultation, and management for the patient. Patients with total or immediate paralysis as well as those with poorly prognostic audiogram results are good candidates for surgical repair. PMID- 16243201 TI - Delayed diagnosis of post-traumatic acute myocardial infarction complicated by congestive heart failure. AB - A 53-year-old man experienced persistent chest pain followed by slight shortness of breath after being hit in the chest by a stranger. Chest X-ray study showed no rib fractures but electrocardiography indicated acute anterior wall myocardial infarction. Echocardiography revealed akinesia in both the interventricular septum and anterior left ventricular wall. Emergency cardiac catheterization demonstrated total occlusion of the proximal left anterior descending coronary artery, 9 h after the event. He was successfully treated with coronary angioplasty and stenting procedures. However, poor left ventricular function was observed 3 months after the event despite medications. We conclude that evaluation for possible myocardial injury should be considered soon after blunt chest trauma for early treatment to improve prognosis. PMID- 16243200 TI - Septic arthritis of the acromioclavicular joint. AB - Septic arthritis of the acromioclavicular (AC) joint is a rare but rapidly destructive etiology of acute shoulder pain. We report a case of septic AC joint in the absence of trauma or intravenous drug use. A diabetic man presented with severe right shoulder pain and was later diagnosed with septic arthritis of the AC joint by magnetic resonance imaging (MRI) and positive blood cultures. Chronic diabetic foot ulcers were the source of the patient's group B streptococcus bacteremia. PMID- 16243202 TI - Clinical decision rules and cervical spine injury in an elderly patient: a word of caution. AB - We report a case of a clinically significant cervical spine fracture in an elderly patient without midline cervical tenderness. Application of the NEXUS rule by the treating physicians ruled out the need for radiography. However, knowledge of the Canadian C-spine rule and clinical judgment prompted obtaining a three-view trauma series of the cervical spine and, when the patient's pain increased, a computed tomography scan of the cervical spine. A type III fracture of the dens was found. In review of the case it was recognized that application of the NEXUS rule for this patient was problematic regarding the assessment of mental status. Specifically, the treating physicians did not strictly adhere to the detailed explanations attached to the NEXUS criteria regarding mental status. Clinicians may wish to preferentially apply the Canadian rule for patients over the age of 64 years. PMID- 16243203 TI - Exercise-unrelated sudden death as the first event of anomalous origin of the left coronary artery from the right aortic sinus. AB - Congenital anomalous origins of the coronary arteries represents a rare but well described cause of myocardial ischemia and sudden death. Left coronary artery (LCA) arising from the right sinus of Valsalva is a rare congenital coronary anomaly that seems to be commonly associated with sudden death in young trained athletes. The possibility of a coronary artery anomaly should always be considered in young individuals with a history of chest pain or syncope, particularly if the episodes are triggered by exercise. We describe a case of congenital LCA anomaly in an asymptomatic 10-year-old girl with no family history of sudden death; no previous unexplained syncopal episodes or exercise-induced symptoms were reported. She experienced a cardiac arrest while she was resting at school and was not recoverable despite early emergency department admission and intensive prolonged cardiopulmonary resuscitation attempts. Post-mortem pathological findings revealed a single origin from the right sinus of Valsalva for both right and left coronary arteries. The LCA was compressed between the aorta and the pulmonary trunk. Histologic features suggested recent ischemia. Although sudden death can be the first manifestation of this condition, it is important to be particularly aware of prodromic symptoms: exertional dyspnea, chest pain, syncope or dizziness. Recognition during life of this coronary anomaly is mandatory to prevent the risk of sudden death and to plan surgical correction if clinically indicated. PMID- 16243204 TI - An ovary with a twist: a case of interesting sonographic findings of ovarian torsion. AB - An otherwise healthy young woman presented to the Emergency Department with abdominal pain. Multiple diagnoses were considered, as must be in cases of women with lower quadrant abdominal pain. Rapid identification of an abnormally enlarged ovary in close proximity to the opposite ovary on pelvic ultrasonography suggested the diagnosis of ovarian torsion. This was verified on laparoscopy. Of note is the fact that normal Doppler flow to both ovaries was demonstrated on pelvic ultrasound. We present this case, and its associated images, with the intent to highlight a readily identifiable sign of ovarian torsion for emergency physicians and to briefly review ovarian torsion's salient clinico-pathologic features. PMID- 16243205 TI - Propofol-induced seizure-like phenomena. AB - Procedural sedation is a common practice in Emergency Medicine. Propofol has supplanted benzodiazepines in many centers as the drug of choice for procedural sedation. This article reports a case of seizure-like activity in an elderly man undergoing procedural sedation for a fracture reduction. The seizure-like activity was attributed to propofol. A review of the literature is discussed. When using propofol for sedation one should be aware of the risk of seizure-like activity. PMID- 16243206 TI - Methylenedioxymethamphetamine (ecstasy)-related hyperthermia. AB - MDMA (or 3, 4 methylenedioxymethamphetamine) was first manufactured in the 1920s and found to have structural similarities to both mescaline and amphetamines. Used briefly by some therapists in the 1970s and early 1980s as an adjunct to psychotherapy, it is now primarily abused by teenagers and young adults as an illicit recreational drug known as "ecstasy." As its popularity has increased, so have the number of fatalities and adverse events related to its use. We report six patients suffering fatal or life-threatening hyperthermia after MDMA use. These cases illustrate that hyperthermia associated with MDMA use cannot be solely attributed to rave parties (high ambient temperatures, excessive dancing, dehydration, and overcrowded conditions), drug contaminants, or co-ingestants. A better understanding of the etiology of hyperthermia after MDMA use is needed so that appropriate harm-reduction measures can be developed and instituted. PMID- 16243207 TI - Accuracy of emergency medicine ultrasound in the evaluation of abdominal aortic aneurysm. AB - This study assesses the accuracy of Emergency Medicine (EM) residents in detecting the size and presence of abdominal aortic aneurysms (AAAs) using EM ultrasound (EUS) compared to radiology measurement (RAD) by computed tomography (CT) scan, magnetic resonance imaging (MRI), angiography, or operative findings. There were 238 aortic EUS performed from 1999-2000; 36 were positive for AAA. The EUS finding of "AAA" had a sensitivity of 0.94 (0.86-1.0 95% confidence interval [CI]) and specificity of 1 (0.98-1.0 95% CI). Mean aortic diameter among patients with AAA identified by EUS was 5.43+/-1.95 cm and by RAD was 5.35+/-1.83 cm. The mean absolute difference between EUS and RAD diameters was 4.4 mm (95% CI 3.7-5.5 mm). Regression of EUS on RAD diameters is strongly correlated, with R(2)=0.92. EM residents with appropriate training can accurately determine the presence of AAA as well as the maximal aortic diameter. PMID- 16243208 TI - Emergency diagnostic paracentesis to determine intraperitoneal fluid identity discovered on bedside ultrasound of unstable patients. AB - Patients presenting with hypotension may be evaluated with a FAST (Focused Abdominal Sonography for Trauma) examination as recent literature has suggested its utility in the unstable patient. Those who are found to have intraperitoneal fluid on the FAST examination may have solid organ injury from unknown trauma, ruptured abdominal aortic aneurysm (AAA), hemorrhaging ovarian cyst, ruptured ectopic pregnancy, or other disease process responsible for intra-periteal blood. However, because ultrasound does not assist in fluid identification, it is possible that the fluid present is not blood, but ascites. Further, a decision may need to be made regarding emergent surgical intervention or blood transfusion vs. aggressive fluid resuscitation or pressors followed by other testing such as computed tomography. This case series illustrates the utility of an ultrasound guided, emergent diagnostic paracentesis in the management of unstable patients found to have a positive FAST examination. Six unstable patients were evaluated with the FAST examination and found to have large quantities of intraperitoneal fluid. In each case the fluid was sampled and proposed management changed due to fluid identification. PMID- 16243209 TI - Pleuritic chest pain and exertional dyspnea. PMID- 16243210 TI - An unusual urethral foreign body: silicone caulk. PMID- 16243211 TI - Coat hanger injury. PMID- 16243212 TI - Third degree heart block in a young man. PMID- 16243213 TI - Subdural hematoma in a shunted patient. PMID- 16243214 TI - Anterior pneumothorax. PMID- 16243215 TI - ED electrical cardioversion for atrial fibrillation: cardiologists are shocked! PMID- 16243216 TI - The "other" primary headaches in children and adolescents. AB - Headache represents one of the most common reasons why children and adolescents are referred to pediatric neurology practices where the most common headache syndromes diagnosed are migraine and its variants, and chronic daily headache. The bulk of recent literature regarding headache in children has focused on these two clinical entities even though large epidemiologic studies have demonstrated that tension-type headache may be two to three times more common in children. Why has so little attention been given to these other disorders? The purpose of this review is to examine the "other" primary headache disorders in children and adolescents. PMID- 16243217 TI - Effectiveness of topiramate in the treatment of pediatric chronic daily headache. AB - This study reports on the efficacy and safety of low-dose topiramate in the treatment of pediatric patients with chronic daily headache. Topiramate is one of the new antiepileptic drugs commonly being used for migraine prophylaxis in adults as well as children and was recently approved by the Food and Drug Administration for migraine treatment in adults. This report presents our experience with low-dose topiramate for the treatment of chronic daily headache using a retrospective parental survey of 21 patients ranging in age from 6 to 18 years. Efficacy and safety were evaluated using a parental assessment and satisfaction questionnaire. Sixty-two percent of families reported that low-dose topiramate (average dose of 30 mg/day) was successful in reducing both the frequency and severity of headache episodes. The headache frequency decreased from 22.8 headaches/month to 7.2 headaches/month and severity decreased from a pain score of 8.1 to 3.7. Topiramate was safe, well tolerated, and highly effective at low doses in the treatment of chronic daily headaches. PMID- 16243218 TI - Comparison of two methods of neurologic assessment in infants. AB - Information provided by the neonatal neurologic assessment is important for identifying infants with neurologic abnormalities at a very early age. The aim of this study was to compare two distinct approaches to the neurologic assessment of newborns: the Amiel-Tison neurologic assessment, and Prechtl's qualitative assessment of general movements. The results of both assessments were studied in a group of 45 preterm infants with different risk factors for brain damage that were compared at term age and at a corrected age of 3 months. The predictive power of the two methods regarding the neurologic and developmental outcome at a corrected age of 12-15 months was analyzed. The agreement of the two methods was excellent at term (kappa = 0.87) and good at 3 months (kappa = 0.54). The sensitivity of both methods for detecting children with neurologic abnormalities was high both at term and at 3 months (0.92, 1.0 for the Amiel-Tison assessment and 0.96, 1.0 for general movements). The specificity of both techniques was low at both ages (0.45, 0.75 for the Amiel-Tison assessment and 0.40, 0.35 for general movements). The agreement of the neurologic and developmental outcome was better with the Amiel-Tison assessment (kappa = 0.39, 0.77) than with the observation of general movements (kappa = 0.38, 0.37). PMID- 16243219 TI - Diffusion tensor imaging in Niemann-Pick Type C disease. AB - Niemann-Pick Type C disease is a homozygous recessive disorder resulting in errant intracellular cholesterol metabolism and the accumulation of intracellular unesterified cholesterol and sphingolipids. Although no current effective treatment exists for Niemann-Pick Type C disease, a number of therapies are under development in animal models. As therapies are brought into clinical trials, it will be extremely helpful to have a reliable means to track the progression of the disease and to monitor its response to therapy. In this effort, diffusion tensor imaging has been applied to investigate the white matter in a Niemann-Pick Type C patient, and the results compared to those from age-matched control subjects. Diffusion tensor imaging enables quantitative measurement of water diffusion in white matter, which is sensitive to the architecture and integrity of the tissue. Compared with control subjects, significant reductions in fractional anisotropy values were observed in regions of white matter, most prominently in the corpus callosum. The results from this case study suggest that diffusion tensor imaging may allow progression of the disease to be quantitatively measured and may be able to play a role as a surrogate marker in clinical trials. PMID- 16243220 TI - Neurotrophic factor expression in three infants with Ondine's curse. AB - This study investigates the expression of some neurotrophic factors (brain derived neurotrophic factor, glial-derived neurotrophic factor, and nerve growth factor) in the cerebrospinal fluid of infants suffering from idiopathic congenital central hypoventilation syndrome and determines their correlations with this syndrome. Cerebrospinal fluid samples were collected from three infants suffering from idiopathic congenital central hypoventilation syndrome and 15 control subjects with obstructive hydrocephalus to measure the expression of brain-derived neurotrophic factor, glial-derived neurotrophic factor, and nerve growth factor using an immunoenzymatic assay. In the cerebrospinal fluid of patients, analysis of neurotrophic factors expression indicated a reduction, not statistically significant, of brain-derived neurotrophic factor compared with the mean level of the control group (1554 pg/mL, 1509 pg/mL, and 1582 pg/mL respectively, in comparison to 1954 +/- 103 pg/mL), whereas nerve growth factor and glial-derived neurotrophic factor did not undergo significant variations in either group. Neurotrophic factors, namely brain-derived neurotrophic factor, regulate the maturation and differentiation of respiratory neurons. The reduced expression of brain-derived neurotrophic factor in the cerebrospinal fluid samples of infants with Ondine's curse, although not statistically significant, is suggestive of a dysregulation in the brain-derived neurotrophic factor synthesis that could play an important role in the breathing disorders observed in patients with idiopathic congenital central hypoventilation syndrome. PMID- 16243221 TI - Oxcarbazepine in infants and young children with partial seizures. AB - In this open-label study, the safety, tolerability, and pharmacokinetics of oxcarbazepine as monotherapy or adjunctive therapy were studied in infants and young children with partial seizures. In a 30-day treatment phase, oxcarbazepine was titrated from 10 mg/kg/day to 60 mg/kg/day. Blood samples for analysis of the oxcarbazepine metabolite, the 10-monohydroxy derivative (MHD), were obtained at regular intervals. Patients completing the treatment phase entered a 6-month extension phase. Safety and tolerability were assessed throughout the study. Twenty-four patients (mean [range] age, 20.4 [2-45] months) were enrolled. Nineteen (79%) patients completed the treatment phase and, together with one patient who discontinued prematurely during the treatment phase, entered the extension phase. Thirteen of 20 (65%) patients completed the extension phase. The most common adverse events were pyrexia, ear infection, and irritability. Whether patients (n = 23) received enzyme-inducing antiepileptic drugs or not, MHD concentrations were consistent with those predicted from a linear, one compartment, population-pharmacokinetic model based on a model previously fitted for 3- to 17-year-old children. Oxcarbazepine was safe and well tolerated in infants and young children. The pharmacokinetic profile of MHD was predicted by extension of a model based on older children. PMID- 16243222 TI - Reactive EEG patterns in pediatric coma. AB - This study was designed to determine whether the observed reactive electroencephalographic patterns in comatose children were associated with a better outcome. All electroencephalograms performed in comatose children aged 2 months to 18 years during the period 1996-2003 were retrospectively analyzed and classified according to modified Young's classification. Reactivity to painful/auditory stimuli and passive eye closure (at least two modalities) was checked in all electroencephalograms. The clinical outcome at 1 year or during the last clinic/inpatient follow-up was scored according to the Pediatric Cerebral and Overall Performance Category Scale. Outcomes were then compared using Fisher exact test and the Mann-Whitney test. Thirty-three patients had electroencephalography within 72 hours after the onset of coma. Fourteen of 33 electroencephalograms revealed reactive patterns. Outcome was unfavorable in 4 (28.6%) of these patients. Three children had no residual neurologic impairment. Among the 19 children with nonreactive electroencephalogram, 13 (65%) had unfavorable outcome, which included 10 deaths. All the survivors had residual neurologic impairment. Outcome was better in children with reactive electroencephalographic patterns (Fisher exact test; P = 0.023). Comatose children with reactive electroencephalographic patterns have better clinical outcome in terms of morbidity and mortality. A careful assessment of electroencephalographic reactive patterns in all comatose children is required for better understanding of the clinical outcome. PMID- 16243223 TI - Macrophagic myofasciitis in childhood: a controversial entity. AB - Macrophagic myofasciitis is an unusual inflammatory myopathy, which has been almost exclusively reported in French adults with diffuse arthromyalgias and asthenia. It is characterized by an infiltrate of densely packed macrophages, with granular periodic-acid-Schiff positive content, on muscle biopsies at the site of vaccination. The presence of aluminum inclusions in these macrophages points to an inappropriate reaction to aluminum used as an adjuvant in some vaccines. Although in adults this entity is well defined, less than 15 cases have been reported in children. This study describes seven children, younger than 3 years of age, with typical lesions of macrophagic myofasciitis on quadriceps muscle biopsy. In five cases, biopsies were performed to exclude mitochondrial pathology. All the children developed hypotonia and motor or psychomotor delay, associated with others symptoms. Abnormal neuroimaging was evident in six cases. Spectrometry studies detected elevated levels of aluminum in muscle in three of four cases tested. Despite the wide use of vaccines in childhood, macrophagic myofasciitis was rarely observed in children and its characteristic histologic pattern could not be correlated with a distinctive clinical syndrome. PMID- 16243224 TI - Cognition in African children with attention-deficit hyperactivity disorder. AB - The aims of the study were: (1) to describe the performance of African children with symptoms of attention-deficit hyperactivity disorder on selected neuropsychologic tests and compare it with performance among peers of the same age without symptoms; (2) to explore through a factor analysis if the selected tests cover the same functions as known from studies in Europe and North America. A nested case-control approach was used to select the two groups of children. The tests were selected to measure aspects of executive functions, attention and memory functions as well as motor skills. A total of 185 schoolchildren (28 cases and 157 control subjects) aged 85 to 119 months old were included. The findings indicate only minor difference between children with symptoms of attention deficit hyperactivity disorder and control subjects in most of the tasks. However, children with symptoms of attention-deficit hyperactivity disorder performed more poorly on tests of motor skills and had more violations of rules on the planning task. The factor analysis indicated a three-factor model, confirming that the selected tests could be used as measures of executive/motor functions, attention, and memory functions. Similar findings have been reported among children in Europe and North America. PMID- 16243225 TI - Delayed, recurrent opsoclonus-myoclonus syndrome responding to plasmapheresis. AB - Opsoclonus-myoclonus syndrome is a distinct neurologic disorder characterized by opsoclonic eye movements, multifocal myoclonus, and ataxia, traditionally described as "dancing eyes, dancing feet." A presenting sign in 2% of children with neuroblastoma, it usually heralds a favorable prognosis for the tumor. Although opsoclonus-myoclonus syndrome usually presents at initial diagnosis or relapse, there are reports of delayed presentation, usually a few months after diagnosis. This report describes a patient with ganglioneuroblastoma who developed recurrent symptoms of opsoclonus-myoclonus syndrome 9 years after completing treatment, without evidence of recurrent tumor. Believed to be autoimmune in origin, opsoclonus-myoclonus syndrome frequently responds to immunomodulatory therapies, such as steroids or intravenous immunoglobulin. This patient did not respond adequately to either agent, so plasmapheresis, a less commonly used modality in opsoclonus-myoclonus syndrome, was attempted. His symptoms resolved after he received therapy with a combination of plasmapheresis and steroids over a 1-year period. After being slowly weaned off all therapy, he has been symptom-free now for over 3 years. Armstrong MB, Robertson PL, Castle VP. Delayed, recurrent opsoclonus-myoclonus syndrome responding to plasmapheresis. PMID- 16243226 TI - Cyclosporine in chronic inflammatory demyelinating polyradiculoneuropathy. AB - This study demonstrates the efficacy of cyclosporine included in a regimen for the treatment of steroid-resistant chronic inflammatory demyelinating polyradiculoneuropathy in two children. Clinical response was characterized by either decreased frequency of recurrent weakness or normalized motor function. Nerve conduction studies and monitoring of cyclosporine levels were included in the serial follow-up evaluations, and their results were used in formulating a treatment plan. One of the two children, who had been monitored for 56 months since the onset of the disease, was able to maintain normal muscle strength without recurrent weakness for 39 months, with 5 mg/kg daily of cyclosporine. The other child, who had been taking prednisolone 0.3 mg/kg daily and cyclosporine 5 mg/kg daily, regained ambulation without support while demonstrating a reduction of recurrent weakness. None had adverse effects caused by cyclosporine therapy. We conclude that cyclosporine is an effective drug in the treatment of children with steroid-resistant chronic inflammatory demyelinating polyradiculoneuropathy. PMID- 16243227 TI - Intrathecal baclofen overdose followed by withdrawal: clinical and EEG features. AB - Intrathecal baclofen therapy is increasingly used to alleviate medically intractable spasticity in children with cerebral palsy, spinal cord injuries, and generalized dystonia. Complications like overdose or withdrawal can occur and could be the result of pump malfunction (device-related) or refilling and programming mistakes (human errors). This report describes a case, with emphasis on electroencephalographic changes, of a 12-year old male on long-term intrathecal baclofen therapy who had sequential occurrence of both acute inadvertent baclofen overdose followed by withdrawal symptoms. During baclofen intoxication, electroencephalography documented periodic generalized epileptiform discharges, occasionally followed by intermittent electro-decremental responses on a background of diffuse delta slowing (1-2 Hz). During withdrawal, mild generalized slowing during wakefulness was observed along with the appearance of high-amplitude, sharply contoured delta activity resembling frontal intermittent rhythmic delta activity in sleep. To our knowledge, this temporal profile of electroencephalographic features during baclofen intoxication followed by withdrawal has not been described before in pediatric patients. It is important for treating physicians to recognize the evolution of this electroencephalographic pattern in order to avoid misinterpretation of diagnosis and prognosis. PMID- 16243228 TI - Extrapyramidal parkinsonism complicating acute organophosphate insecticide poisoning. AB - The aim of this study is to report our experience with a child who developed extrapyramidal perturbations complicating acute organophosphate insecticides poisoning and to review the literature reporting on basal ganglia impairment associated with this poisoning. Our patient had developed overt parkinsonism presenting with a resting tremor, expressionless face, and lack of blinking along with marked cogwheel rigidity and a stooped, slow gait. He was alert, coherent, and cooperative, yet agitated. The parkinsonian perturbations developed 5 days after an accidental ingestion of a raw eggplant sprayed with the organophosphate dimethoate (Rogor) when he had already recovered from the acute cholinergic crisis, the first stage of organophosphate poisoning. Such a presentation was initially perceived by his caregivers as severe reactive depression or even psychosis. Once a parkinsonian syndrome was diagnosed, he was begun on amantadine and completely recovered within 1 week with no relapse of symptoms. Basal ganglia impairment should be considered in any patient who develops extrapyramidal symptoms such as marked rigidity and bradykinesia or choreoathetosis while recovering from the acute cholinergic phase of organophosphate insecticide poisoning. Thus, administration of a drug such as amantadine, which probably enhances neurotransmission, may hasten the rate of recovery and prevent long-term neurologic and emotional sequelae. PMID- 16243229 TI - Antibiotic resistance in outpatient urinary isolates: final results from the North American Urinary Tract Infection Collaborative Alliance (NAUTICA). AB - The goal of the North American Urinary Tract Infection Collaborative Alliance (NAUTICA) study was to determine antibiotic susceptibility to commonly used agents for urinary tract infections against outpatient urinary isolates obtained in various geographic regions in the USA and Canada. Forty-one medical centres (30 from the USA and 11 from Canada) participated, with each centre submitting up to 50 consecutive outpatient midstream urine isolates. Isolates were identified to species level by the standard protocol of each laboratory. Susceptibility testing was determined using the National Committee for Clinical Laboratory Standards (NCCLS) microdilution method. Resistance breakpoints used were those published by the NCCLS, including: ampicillin (resistant > or = 32 microg/mL), sulphamethoxazole/trimethoprim (SMX/TMP) (resistant > or = 4 microg/mL), nitrofurantoin (resistant > or = 128 microg/mL), ciprofloxacin (resistant > or = 4 microg/mL) and levofloxacin (resistant > or = 8 microg/mL). Of the 1990 isolates collected, 75.1% (1494) were collected from the USA and 24.9% (496) were collected from Canada. The mean age of the patients was 48.3 years (range 1 month to 99 years), and 79.5% and 20.5% of isolates were obtained from women and men, respectively. The most common organisms were Escherichia coli (57.5%), Klebsiella pneumoniae (12.4%), Enterococcus spp. (6.6%), Proteus mirabilis (5.4%), Pseudomonas aeruginosa (2.9%), Citrobacter spp. (2.7%), Staphylococcus aureus (2.2%), Enterobacter cloacae (1.9%), coagulase-negative staphylococci (1.3%), Staphylococcus saprophyticus (1.2%), Klebsiella spp. (1.2%), Enterobacter aerogenes (1.1%) and Streptococcus agalactiae (1.0%). Among all 1990 isolates, 45.9% were resistant to ampicillin, 20.4% to SMX/TMP, 14.3% to nitrofurantoin, 9.7% to ciprofloxacin and 8.1% to levofloxacin. Fluoroquinolone resistance was highest in patients > or = 65 years of age. For the 1142 E. coli isolates, resistance rates were: ampicillin 37.7%, SMX/TMP 21.3%, ciprofloxacin 5.5%, levofloxacin 5.1% and nitrofurantoin 1.1%. For all 1990 isolates and for the 1142 E. coli only, resistance rates were significantly higher in US compared with Canadian medical centres. This study reports higher rates of antibiotic resistance in US versus Canadian outpatient urinary isolates and demonstrates the continuing evolution of resistance to antimicrobial agents. PMID- 16243230 TI - Mathematical model for carbon dioxide evolution from the thermophilic composting of synthetic food wastes made of dog food. AB - The impacts of the aeration and the agitation on the composting process of synthetic food wastes made of dog food were studied in a laboratory-scale reactor. Two major peaks of CO(2) evolution rate were observed. Each peak represented an independent stage of composting associated with the activities of thermophilic bacteria. CO(2) evolutions known to correlate well with microbial activities and reactor temperatures were fitted successfully to a modified Gompertz equation, which incorporated three biokinetic parameters, namely, CO(2) evolution potential, specific CO(2) evolution rate, and lag phase time. No parameters that describe the impact of operating variables are involved. The model is only valid for the specified experimental conditions and may look different with others. The effects of operating parameters such as aeration and agitation were studied statistically with multivariate regression technique. Contour plots were constructed using regression equations for the examination of the dependence of CO(2) evolution potentials on aeration and agitation. In the first stage, a maximum CO(2) evolution potential was found when the aeration rate and the agitation parameter were set at 1.75 l/kg solids-min and 0.35, respectively. In the second stage, a maximum existed when the aeration rate and the agitation parameter were set at 1.8 l/kg solids-min and 0.5, respectively. The methods presented here can also be applied for the optimization of large scale composting facilities that are operated differently and take longer time. PMID- 16243231 TI - Country report: Broiler industry and broiler litter-related problems in the southeastern United States. AB - This report describes the development of the broiler litter problem in the southeastern United States, including the economic opportunity and environmental challenges brought to the region by the industry. Through an analysis applied to the State of Georgia, land application of litter as a disposal alternative is examined along with its associated benefits. The analysis indicates that litter could be transported economically up to 256 km for cropland application. Excessive broiler litter production in a few concentrated regions is expected to stimulate the development of alternative approaches to broiler litter management, such as electricity generation. PMID- 16243232 TI - Osteoblasts from the sclerotic subchondral bone downregulate aggrecan but upregulate metalloproteinases expression by chondrocytes. This effect is mimicked by interleukin-6, -1beta and oncostatin M pre-treated non-sclerotic osteoblasts. AB - OBJECTIVE: To determine the effects of osteoarthritic (OA) subchondral osteoblasts on the metabolism of human OA chondrocytes in alginate beads. METHODS: Human chondrocytes were isolated from OA cartilage and cultured in alginate beads for 4 days in the absence or in the presence of osteoblasts isolated from non-sclerotic (N) or sclerotic (SC) zones of human OA subchondral bone in monolayer (co-culture system). Before co-culture, osteoblasts were incubated for 72 h with or without 1.7ng/ml interleukin (IL)-1beta, 100 ng/ml IL 6 with its soluble receptor (50 ng/ml) or 10 ng/ml oncostatin M (OSM). Aggrecan (AGG) and matrix metalloproteases (MMP)-3 and -13 mRNA levels in chondrocytes were quantified by real-time polymerase chain reaction. AGG production was assayed by a specific enzyme amplified sensitivity immunoassay. RESULTS: SC, but not N, osteoblasts induced a significant inhibition of AGG production and AGG gene expression by human OA chondrocytes in alginate beads, and significantly increased MMP-3 and MMP-13 gene expression by chondrocytes. When they were pre incubated with IL-1beta, IL-6 or OSM, N osteoblasts inhibited AGG synthesis and increased MMP-3 and -13 gene expression by chondrocytes in alginate beads in a same order of magnitude as SC osteoblasts. CONCLUSIONS: These results demonstrate that SC OA subchondral osteoblasts could contribute to cartilage degradation by stimulating chondrocytes to produce more MMP and also by inhibiting AGG synthesis. PMID- 16243233 TI - Mini dental implants: an adjunct for retention, stability, and comfort for the edentulous patient. AB - OBJECTIVE: This article examines the success of mini-dental implants (MDI'S) by assessing four subjective measures of patient satisfaction for MDI's in the edentulous maxilla and mandible: comfort, retention, chewing ability and speaking ability. Success rates, surgical techniques, and financial advantages of the MDI's are reviewed. STUDY DESIGN: Thirty consecutive patients received four MDI's between the mental foramen of the mandible from 9/18/2003 to 10/22/2004. Questionnaires were sent to all thirty patients an average of 5 months postoperatively. The patients ranked comfort, retention, chewing ability, and speaking ability from 1 to 10 (1=poor and 10=excellent). RESULTS: A total of 116 MDI's were placed in 13 months and 113 remain stable for a 97.4% implant success rate. Pre-operatively patients rated their retention at 1.7+/-0.42 and post operatively at 9.6+/-0.37, for a difference of 7.9 (p=3.6-19). Comfort was the next greatest improvement, with a pre-operative rating of 2.2+/-0.63 and a post operative rating of 9.4+/-0.45, for a difference of 7.2 (p=3.5-15). Chewing ability also improved, with a difference of 7.0 (p=2.9e-16). In the final category of speaking ability, the pre-operative to post-operative difference was 3.2 (p=1.1e-5). CONCLUSION: MDI's are a highly successful implant option for patients with poor tolerance to maxillary and mandibular prosthesis. The implants are relatively affordable and overall patient satisfaction is excellent. PMID- 16243234 TI - Mandibular midline distraction using a simple device. AB - OBJECTIVE: Mandibular midline distraction osteogenesis represents a new option for creation of intra-arch space in the mandibular arch. The aim of this paper is to introduce a simple device (hyrax expander) and method for mandibular midline distraction. STUDY DESIGN: The sample consisted of 24 patients (mean age: 18.07 years) treated with mandibular midline distraction. Hyrax expanders were used as symphyseal distractors, and distraction procedure was carried out with a rate of 1 mm and rhythm of twice daily. RESULTS: Mandibular midline distraction was completed successfully in all patients and the distraction amount was 7.01 mm. No major complication other than mild mucosal irritation and gingival recession was observed. CONCLUSION: The presented method is a viable option for mandibular midline distraction osteogenesis. PMID- 16243235 TI - The world is flat. PMID- 16243236 TI - Trigeminal neuralgia. AB - Trigeminal Neuralgia (TN) is a well recognized syndrome characterized by lancinating attacks of severe facial pain. The diagnosis of TN is based on a history of characteristic pain attacks that are consistent with specific widely accepted criteria for the diagnosis. TN pain attacks may result from physiologic changes induced by a chronic partial injury to the brainstem trigeminal nerve root from a variety of causes. An early and accurate diagnosis of TN is important, because therapeutic interventions can reduce or eliminate pain attacks in the large majority of TN patients. PMID- 16243237 TI - Nasal airflow and olfactory function after the repair of cleft palate (with and without cleft lip). AB - OBJECTIVE: The objective of this study was to examine nasal airflow and olfactory functions in patients with repaired cleft palate compared with matching normal controls. STUDY DESIGN: The all-cleft group consisted of 25 patients with hard palate cleft comprising 15 patients with unilateral cleft palate and lip (UCLP); 2 with CP but no cleft lip (UCLP subgroup) and 8 patients with bilateral cleft lip and palate (BCLP subgroup). All had had surgical correction of the palate in infancy. The control group consisted of 20 nonaffected orthodontic patients. The median age of both groups was 14 years. The tests included the following: (1) nasal airflow measured by anterior rhinomanometry, (2) smell threshold for isoamyl-acetate determined using a 3-way forced choice method, (3) a self administered questionnaire regarding the subjective perception of smell sense function, and (4) orthonasal and retronasal smell identification (correct/incorrect) and hedonics using visual analog scale (VAS). RESULTS: The respective test results follow. (1) When compared with the control group, the total airflow in the UCLP subgroup was significantly lower especially on the affected side; while in the BCLP subgroup it was lower than in the control group bilaterally. No significant difference was found between the cleft side of UCLP and BCLP subgroups. (2) The smell threshold of the UCLP subgroup was significantly higher than that of the control group and BCLP subgroup. No significant differences were found between right and left nostrils within the BCLP patients and between them and the control group. (3) No difference was found between the groups regarding the subjective perception of smell. (4) No significant differences were found between the UCLP and BCLP subgroups and between the all-cleft group and the control group, except for one item, regarding orthonasal and retronasal smell identification and hedonics. CONCLUSION: Although nasal airflow is significantly lower and the smell threshold higher on the cleft side, the day-to-day function of the sense of smell of cleft patients is similar to that of normal controls. PMID- 16243238 TI - Radiographic proximity of the mandibular third molar to the inferior alveolar canal. AB - OBJECTIVE: To determine the panoramic radiographic distance from the mandibular third molar tooth to the inferior alveolar canal. STUDY DESIGN: Five hundred sixty mandibular third molars were evaluated by panoramic radiography. The teeth were grouped into erupted vs unerupted and further subdivided by tooth angulation. The distance from the most inferior aspect of the mandibular third molar tooth to the superior border of the inferior alveolar canal was measured with digital calipers. A t test was performed to compare erupted and unerupted teeth, and ANOVA was used to determine if a significant difference exists based upon tooth angulation. A record review was performed to assess the incidence of inferior alveolar nerve paresthesia based upon measured distances. RESULTS: The mean distance from erupted mandibular third molar teeth to the inferior alveolar canal is 0.88 mm. This distance was significantly different from unerupted teeth (P=.002). The mean values for unerupted teeth indicated that the most inferior portion of all teeth measured was below the superior border of the canal (negative values) as follows: mesioangular (-0.97 mm), vertical (-0.61 mm), distoangular (-0.31 mm), and horizontal (-0.24 mm). The position of mesioangular impactions were significantly different than all other impaction groups (P=.0125). The incidence of inferior alveolar nerve paresthesia was 3.33% (18/541), most commonly associated with mesioangular impactions (-0.66 mm) in female patients. CONCLUSIONS: Unerupted mandibular third molar teeth are closer to the inferior alveolar canal than are erupted teeth. Mesioangular mandibular third molar impactions are most closely positioned to the inferior alveolar canal, and this may represent an independent risk factor for postoperative paresthesia. PMID- 16243239 TI - Clindamycin in dentistry: more than just effective prophylaxis for endocarditis? AB - Clindamycin is a broad-spectrum antibiotic with activity against aerobic, anaerobic, and beta-lactamase-producing pathogens. This antibiotic has been used for many years as prophylactic treatment during dental procedures to prevent endocarditis. However, the spectrum and susceptibility of the bacteria species involved in dental infections indicate that clindamycin would also be an effective treatment option for these conditions. In addition to its antiinfective properties, clindamycin has high oral absorption, significant tissue penetration, including penetration into bone, and stimulatory effects on the host immune system. This review discusses the microbiologic and clinical evidence supporting the efficacy and safety of clindamycin for the successful management of dental infections. PMID- 16243240 TI - The effects on postoperative oral surgery pain by varying NSAID administration times: comparison on effect of preemptive analgesia. AB - OBJECTIVE: Many studies on the efficacy of preemptive analgesia have been processed in different ways. But the value of preemptive analgesia is still controversial. The goal of this study was to compare analgesic effects of a nonsteroidal anti-inflammatory drug (NSAID) for oral surgical pain according to 3 different administration times. STUDY DESIGN: Using a randomized, parallel-group, single-center, and active-controlled test design, this study was conducted with 80 healthy patients undergoing a surgical removal of an impacted mandibular third molar requiring bone removal. The oral NSAID was first administered 1 hour preoperatively, or 1 hour postoperatively, or no scheduled administration pre- or postsurgery. Whenever patients felt at least moderate pain (score > or =5 on a 10 point scale) after surgery, they were instructed to take the same drug. Pain intensities and times to the first and second onsets of postoperative pain from the end of surgery were assessed for 24 hours. RESULTS: Of the 80 enrolled subjects in this study, 25 patients were assigned to the preemptive group, 26 to the posttreatment group, and 29 to the no-treatment group. The demographic distribution and duration of surgery in the 3 groups were statistically similar. The mean time to first onset of postoperative pain was significantly prolonged in the posttreatment group (277.2 minutes, P < .05) compared to the preemptive group (158.4 minutes) and the no-treatment group (196.5 minutes). The mean time to second onset of postoperative pain was not significantly different among the 3 groups. No significant statistical difference was found among the mean pain intensities at the first and second onsets of postoperative pain in the 3 groups. CONCLUSIONS: In this small selected group of subjects and limited study design, the analgesic effects of NSAID administered preoperatively were no longer effective for postoperative pain. The results in this population imply that scheduled postoperative analgesics before pain development are adequate for postoperative analgesia without preoperative administration. PMID- 16243241 TI - Midazolam in the reduction of surgical stress: a randomized clinical trial. AB - OBJECTIVE: The objective of this study was to assess the role of midazolam in reducing surgical stress as measured using subjective and objective variables. STUDY DESIGN: The study was a double-blind randomized controlled trial. Thirty eight male patients undergoing surgical removal of third molars under general anesthesia were recruited for this study, each patient was given premedication (midazolam or placebo) and subjective variables (HAD scale) were obtained and objective variables (salivary cortisol samples and vital signs) were collected pre-, peri-, and postoperatively. The salivary samples were analyzed by direct immunofluorimetric assay using the "DELFIA" system. RESULTS: There were no significant differences in anxiety between the treatment group and the control group before the administration of the premedication. Following the administration of premedication, the majority of the control group showed high cortisol levels on the day of surgery, compared with relatively low cortisol levels in the majority of the treatment group. A few patients in the control group gave a placebo effect (sedative effect) and a number of the treatment group were unresponsive to the drug. There was a slight drop in the blood pressure and respiration rate with a slight increase in the heart rate in the treatment group; however these results were not statistically significant. The HAD scores were not statistically different between the 2 groups. CONCLUSION: Midazolam has proved to be very successful in reducing anxiety and stress pre-, peri-, and postoperatively with no significant effect on the vital signs of a healthy patient. Salivary cortisol technique is an easy, noninvasive method to assess anxiety and stress level in patients undergoing surgery. PMID- 16243242 TI - Oral malignancies in HIV disease: changes in disease presentation, increasing understanding of molecular pathogenesis, and current management. AB - Infection with human immunodeficiency virus (HIV) and progression to acquired immune deficiency syndrome (AIDS) are associated with a vide variety of morbidities. Local and systemic diseases can develop in association with HIV infection and may manifest themselves as malignancies of the oropharynx. Advances in HIV management, fueled by increasing understanding of molecular pathogenesis, have resulted in marked changes in the prevalence of oral malignant disease. This paper discusses recent trends in the presentation and treatment of malignancies related to HIV and AIDS with an emphasis on malignancies seen in the oral cavity. PMID- 16243243 TI - Sclerotherapy of benign oral vascular lesion with ethanolamine oleate: an open clinical trial with 30 lesions. AB - OBJECTIVE: The objective of this study was to report and discuss the results from treatment of benign oral vascular lesions with ethanolamine oleate. STUDY DESIGN: Twenty-seven patients with 30 examples of oral vascular malformation, hemangioma, or varix were treated with intralesional injections of 1.25% or 2.5% ethanolamine oleate at an interval of 15 days between each application. The lesions were divided into 2 categories: (1) lesions of 20 mm or less and (2) those greater than 20 mm. Subsequently, the Mann-Whitney test was used a means of statistical analysis. RESULTS: Although the number of injections varied from patient to patient, all lesions responded to the treatment, showing total clinical regression. Lesions of 20 mm or less needed a lesser number of applications than those greater than 20 mm (P < .05). CONCLUSIONS: Ethanolamine oleate is a 100% effective sclerosant agent for treatment of benign oral vascular lesions. In this study, no difference was found between the 2 concentrations applied. PMID- 16243244 TI - External cervical root resorption involving multiple maxillary teeth in a patient with hereditary hemorrhagic telangiectasia. AB - Hereditary hemorrhagic telangiectasia (HHT) is an inherited syndrome characterized by mucocutaneous telangiectases that commonly involve the tongue, lips, fingers, and conjunctiva. While root resorption has been reported in association with central hemangiomas of bone, the association of HHT with external cervical root resorption has not been described to date. We report a case of a 57-year-old female with HTT who presented with advanced cervical root resorption involving multiple maxillary anterior teeth. Histologic examination of the gingival tissue adjacent to the area of root resorption demonstrated multiple thin-walled vascular elements as well as larger vascular channels surrounded by a thickened muscular layer. We hypothesize that the external root resorption seen in this case is the result of the HHT-related vascular process in the adjacent gingival tissue. PMID- 16243245 TI - Peripheral primitive neuroectodermal tumor associated with the anterior mandible: a case report and review of the literature. AB - Neuroectodermal tumors may arise in many places throughout the body including the diverse tissues of the head and neck. The primitive neuroectodermal tumor is a predominately neural, nonepithelial neoplasm similar to Ewing sarcoma. This article describes an 18-year-old female patient with a highly malignant peripheral primitive neuroectodermal tumor located in the soft tissue anterior to the mandibular symphysis. The clinical and radiographic presentation as well as the histopathology and immunohistochemistry of this rare entity is discussed. A review of the literature with respect to this tumor, as well as the current management of this tumor, is presented. PMID- 16243246 TI - Benign teratoma of the buccal mucosa in a 9-year-old girl: report of case and review of the literature. AB - Teratomas are embryonal tumors composed of tissues from all 3 germinal layers with variable levels of maturity. Teratomas are rare in the head and neck region, representing less than 5% of all cases. In the head and neck the most common involved sites are the nasopharynx and cervical regions. Oral teratomas are extremely uncommon, and we found only 21 cases published in the English-language literature. We describe an unusual case of oral teratoma on the right buccal mucosa of a 9-year-old girl. PMID- 16243247 TI - Physical properties and ease of operation of a wireless intraoral x-ray sensor. AB - OBJECTIVE: A wireless CMOS (CDR Wireless) system was evaluated based on its physical properties and ease of operation. STUDY DESIGN: The physical properties were assessed by dose-response curve, modulation transfer function, and detective quantum efficiency tests. The range of signal receptivity between the sensor and the antenna was also determined. The time required to make a radiograph was measured. Using a visual analog scale (VAS), 10 test patients were asked to evaluate the discomfort caused by having the sensor inserted and placed in the mouth. An intraoral x-ray film packet and a wired CDR sensor were also evaluated for comparison with the wireless system. RESULTS: The physical properties of the wireless system and the wired CDR sensor were essentially equal. The antenna could receive the signal up to 3.5 m from the sensor. The wireless CMOS system required less time to perform the radiographic task than the other methods studied. The VAS scores for the wireless and wired CDR sensors were comparable. CONCLUSIONS: The CDR Wireless sensor has equivalent physical properties to its wired counterpart and may be more convenient to use. PMID- 16243248 TI - Three-dimensional identification of hemangiomas and feeding arteries in the head and neck region using combined phase-contrast MR angiography and fast asymmetric spin-echo sequences. AB - OBJECTIVE: To evaluate a proposed technique for the 3-dimensional (3D) detection of hemangiomas, including vascular malformation and their feeding arteries, in the head and neck. The new technique combines phase-contrast magnetic resonance angiography (PCMRA) without contrast medium and 3D fast asymmetric spin-echo (FASE) sequences. METHODS: The technique was applied to 3 patients having hemangiomas in the head and neck region. In 1 patient the image obtained with the proposed technique was compared to that obtained by standard contrast angiography. RESULTS: In all 3 patients, the 3D presence of the hemangiomas and the feeding arteries were well defined in images created by the proposed technique. Additionally, the characterization of the hemangioma's 3D structure and distribution of the feeding arteries coincided with those observed using contrast angiography in the case for which contrast angiography was also performed. CONCLUSIONS: Preliminary experience shows that the proposed technique combining 3D-FASE and 3D-PCMRA is useful to visualize both the 3D structure of hemangiomas and to identify the 3D distribution of the feeding arteries without using contrast medium. PMID- 16243250 TI - Topographic metabolic map of head and neck squamous cell carcinoma using 18F-FDG PET and CT image fusion. AB - OBJECTIVE: To propose a methodologic approach to evaluate head and neck tumors in order to identify and distinguish areas of higher metabolic activity inside the lesion. STUDY DESIGN: The sample consisted of 17 patients with squamous cell carcinoma of the head and neck. Images were simultaneously acquired using a nondedicated PET-CT device and an independent workstation with ENTEGRA 2 NT software to generate the image fusion between PET and CT. Sites of higher metabolic activity inside the tumor were classified as centric or eccentric according to their relative location to the lesion center. RESULTS: Seventy-seven percent (n=13) of the patients presented the site of higher metabolic activity at the center of lesion, and in 23% (n=4) the uptake of the tracer was increased at the periphery of the lesion. CONCLUSIONS: This technique gave a realistic view of the functional metabolism, locating the anatomical tumor area and helping in future treatment planning. PMID- 16243249 TI - Intraoperative imaging techniques: a guide to retrieval of foreign bodies. AB - Foreign bodies are frequently introduced into the tissues of the head and neck by various mechanisms, and oral and maxillofacial surgeons are often called upon to retrieve these embedded objects. Retrieval may be quite challenging depending on many factors such as the size of the object, the location, and the surrounding anatomical structures. Preoperative imaging is very important in deciding upon the surgical approach. Computerized tomography is considered the gold standard for detection of foreign bodies because of the ability to localize an object in multiple planes and the creation of a 3-dimensional image. Difficulty arises when looking for a small object in an area with multiple important anatomical structures, such as the infratemporal fossa or the neck. Surgery can become tedious secondary to the risk of postoperative morbidity with injury to various anatomical structures. Foreign bodies in the head and neck are often difficult to manage even when a plan has been formulated from static preoperative images. Intraoperative feedback or guidance, especially when navigating through troublesome locations, can be extremely useful. In this paper, we report 2 cases and discuss the various modalities used for intraoperative imaging as a guide for surgical retrieval of foreign bodies. PMID- 16243251 TI - The effect of calcium chelating or binding agents on Candida albicans. AB - OBJECTIVE: The aim of this study was to evaluate antifungal effects of calcium chelating or -binding agents on Candida albicans comparing with conventional antifungal agents. STUDY DESIGN: Two clinical oral isolates and 1 standard strain of C albicans were included in the study. Test solutions were ethylenediamine tetraacetic acid (EDTA), ethyleneglycol-tetraacetic acid (EGTA), sodium fluoride (NaF), titanium tetrafluoride (TiF4), nystatin, and ketoconazole. Minimum inhibitory and fungicidal concentrations of the solutions were determined. The results were analyzed statistically using Friedman's nonparametric 2-way ANOVA and Mann-Whitney U tests. RESULTS: All isolates demonstrated similar susceptibility patterns (P>.05). Except ketoconazole, EDTA had the highest antifungal and fungicidal activity, followed by TiF4. EGTA and NaF were the weakest agents against C albicans among all test solutions. CONCLUSION: EDTA and TiF4 may be recommended as an alternative irrigating solution particularly in persistent root canal infections and in root canals of patients having a high incidence of oral candidosis. PMID- 16243252 TI - Radiographic evaluation of periapical status and prevalence of endodontic treatment in an adult Japanese population. AB - OBJECTIVES: To determine the prevalence of periapical radiolucencies and endodontic treatment in an adult Japanese population. STUDY DESIGN: Periapical status and length of root fillings of 672 adult patients attending Okayama University Hospital of Dentistry were evaluated using full mouth intraoral radiographs. RESULTS: Overall, 87% of the subjects had root-filled teeth, and 70% exhibited an apical radiolucency. Of the 16,232 teeth examined, 21% had been root filled, and, of these, 40% exhibited an apical radiolucency. Root-filled teeth that were overfilled or that were mandibular incisors had the highest prevalence of apical radiolucencies. CONCLUSIONS: The prevalence of root-filled teeth appears higher in this Japanese population than in Europe or America; however, the ratio of teeth with an apical radiolucency to root-filled teeth was within the range of that reported for other countries. Overfilled teeth and mandibular incisors are most likely to exhibit apical radiolucencies. PMID- 16243253 TI - Comparative evaluation of the preparation efficacies of HERO Shaper and Nitiflex root canal instruments in curved root canals. AB - OBJECTIVES: To comparatively evaluate the shaping efficacies of HERO Shaper rotary instruments and Nitiflex hand files. STUDY DESIGN: The mesial roots of 40 mandibular molars were used. In 20 teeth, the mesiobuccal canals were instrumented with HERO Shaper and the mesiolingual canals with Nitiflex. In the remaining 20 teeth, the mesiobuccal canals were instrumented with Nitiflex and the mesiolingual canals with HERO Shaper. Pre- and postoperative sections were obtained from the coronal, middle, and apical portions and analyzed. Statistical analysis was performed using the Student t test. RESULTS: More dentin was removed from the middle portion with HERO Shaper (P < .05). No statistically significant difference was observed in terms of transportation (P>.05). No instrument fracture or deformation was noted. CONCLUSIONS: Both HERO Shaper and Nitiflex can be recommended for clinical practice. Further studies can focus on the comparison of HERO Shaper with other rotary instruments as there is an increasing trend for the utilization of these systems. PMID- 16243254 TI - The lost science of formulation. PMID- 16243255 TI - Specific enzyme identified that snips APP into beta-amyloid. PMID- 16243256 TI - Novel alternative for osteoarthritis. PMID- 16243257 TI - Cathepsin enzyme provides clue to SARS infection. PMID- 16243258 TI - Steve Carney talks to Vincent Lee on the pharma industry, the FDA and public education with respect to drugs and their development. PMID- 16243259 TI - Widening opportunities in restenosis. PMID- 16243260 TI - Trends in genomic variation: a view of some of the latest technologies. PMID- 16243261 TI - Scaling the ladder. PMID- 16243262 TI - Keynote review: in vitro safety pharmacology profiling: an essential tool for successful drug development. AB - Broad-scale in vitro pharmacology profiling of new chemical entities during early phases of drug discovery has recently become an essential tool to predict clinical adverse effects. Modern, relatively inexpensive assay technologies and rapidly expanding knowledge about G-protein coupled receptors, nuclear receptors, ion channels and enzymes have made it possible to implement a large number of assays addressing possible clinical liabilities. Together with other in vitro assays focusing on toxicology and bioavailability, they provide a powerful tool to aid drug development. In this article, we review the development of this tool for drug discovery, its appropriate use and predictive value. PMID- 16243263 TI - The role of nanobiotechnology in drug discovery. AB - The application of nanotechnology in life sciences, nanobiotechnology, is already having an impact on diagnostics and drug delivery. Now, researchers are starting to use nanotechnology in the field of drug discovery. This review explains how several technologies, including nanoparticles and nanodevices such as nanobiosensors and nanobiochips, are used to improve drug discovery and development. Nanoscale assays can contribute significantly to cost-saving in screening campaigns. In addition, some nanosubstances (such as fullerenes) could be potential drugs for the future. Although there might be some safety concerns with respect to the in vivo use of nanoparticles, studies are in place to determine the nature and extent of adverse events. Future prospects for the application of nanotechnology in healthcare and for the development of personalized medicine appear to be excellent. PMID- 16243264 TI - High throughput P450 inhibition screens in early drug discovery. AB - This review of high throughput (HT) P450 inhibition technologies and their impact on early drug discovery finds the field at a mature stage. The relationship between P450 inhibition and drug-drug interactions is well understood. A wide variety of P450 inhibition detection technologies are readily available off-the shelf, but what seems still to be missing is a general agreement on how much weight one should give to the various types of early discovery HT P450 inhibition data. Method-dependent potency differences are a cause of concern, and to resolve this issue the authors advocate calibration of the HT methods with a large set of marketed drugs. PMID- 16243265 TI - PEGylation, successful approach to drug delivery. AB - PEGylation defines the modification of a protein, peptide or non-peptide molecule by the linking of one or more polyethylene glycol (PEG) chains. This polymer is non-toxic, non-immunogenic, non-antigenic, highly soluble in water and FDA approved. The PEG-drug conjugates have several advantages: a prolonged residence in body, a decreased degradation by metabolic enzymes and a reduction or elimination of protein immunogenicity. Thanks to these favorable properties, PEGylation now plays an important role in drug delivery, enhancing the potentials of peptides and proteins as therapeutic agents. PMID- 16243266 TI - Can smooth muscle represent a useful target for the treatment of rapid ejaculation? AB - Rapid ejaculation is probably the most common form of male sexual dysfunction. Current research into the treatment of the condition has focused on centrally acting or topical desensitizing agents; however, no treatment has yet been approved. An alternative approach could be to develop drugs that act directly upon the target organ itself and our increasing knowledge of the molecular biology of the accessory sex organs makes this a realistic possibility. This review analyzes the information in the literature that would support such a hypothesis. Particular emphasis has been placed on articles that have investigated smooth muscle cell relaxation. A critical review of the literature has revealed that there are potentially a myriad of targets through which rapid ejaculation can be treated. PMID- 16243267 TI - Investigating Wnt signaling: a chemogenomic safari. AB - Chemical genomics is a powerful method to complement more traditional genetic techniques (i.e. knockout mice, siRNA) for the dissection of complex signaling networks. Wnt signaling in mammals is a complex and crucial regulator of diverse functions. The Wnt-beta-catenin pathway initiates a signaling cascade that is crucial in both normal development and the initiation and progression of cancer. A key step in Wnt activation of target genes is the nuclear translocation of beta catenin and the formation of a complex between beta-catenin and members of the T cell factor (TCF) family of transcription factors. Using a forward chemical genomics strategy, we identified ICG-001, a selective inhibitor of a subset of Wnt-beta-catenin-driven gene expression. This chemogenomic tool enables us to dissect this complex signaling network and to better understand the role of Wnt signaling in both normal and pathophysiological settings. PMID- 16243268 TI - Beyond annotation transfer by homology: novel protein-function prediction methods to assist drug discovery. AB - Every entirely sequenced genome reveals 100 s to 1000 s of protein sequences for which the only annotation available is 'hypothetical protein'. Thus, in the human genome and in the genomes of pathogenic agents there could be 1000 s of potential, unexplored drug targets. Computational prediction of protein function can play a role in studying these targets. We shall review the challenges, research approaches and recently developed tools in the field of computational function-prediction and we will discuss the ways these issues can change the process of drug discovery. PMID- 16243270 TI - Functional effects of Japanese style fermented soy sauce (shoyu) and its components. AB - The functional effects of Japanese style fermented soy sauce (shoyu) have been studied. Soy sauce promotes digestion, because the consumption of a cup of clear soup containing soy sauce enhances gastric juice secretion in humans. Soy sauce possesses antimicrobial activity against bacteria such as Staphylococcus aureus, Shigella flexneri, Vibrio cholera, Salmonella enteritidis, nonpathogenic Escherichia coli and pathogenic E. coli O157:H7. Soy sauce also contains an antihypertensive component. An angiotensin I-converting enzyme inhibitor having antihypertensive effects was found in soy sauce. The active compound was identified as nicotianamine, which comes from soybeans. Soy sauce exhibits anticarcinogenic effects. Giving diets containing soy sauce to mice inhibit benzo[a]pyrene (BP)-induced forestomach neoplasia. The anticarcinogenic compounds in soy sauce were identified. The flavor components of Japanese style fermented soy sauce, such as 4-hydroxy-2(or 5)-ethyl-5(or 2)-methyl-3(2H)-furanone (HEMF), which is a characteristic flavor component of Japanese style fermented soy sauce and 4-hydroxy-2,5-dimethyl-3(2H)-furanone (HDMF) and 4-hydroxy-5-methyl-3(2H) furanone (HMF) exhibit antioxidant activities and anticarcinogenic effects on BP induced mice forestomach neoplasia when fed following carcinogen exposure. The feeding of a diet containing 10% soy sauce to male C3H mice for 13 months also reduces the frequency and multiplicity of spontaneous liver tumors. HDMF and HEMF also exhibit anticataract effects in the spontaneous cataract rat (ICR/f rat). Fermented soy sauce contains three tartaric isoflavone derivatives called shoyuflavones. These shoyuflavones were shown to have inhibitory activities against histidine decarboxylase, which produces histamine, a mediator of inflammation, allergy and gastric acid secretion. Soy sauce also exhibits antiplatelet activity. beta-Carbolines were isolated from soy sauce as the active compounds. Soybeans and wheat, which are the main raw materials of soy sauce, are allergenic foods. However, recent studies by enzyme-linked immunosorbent assay showed the absence of soybean and wheat allergens in soy sauce. PMID- 16243271 TI - Bioreactor design for tissue engineering. AB - Bioreactor systems play an important role in tissue engineering, as they enable reproducible and controlled changes in specific environmental factors. They can provide technical means to perform controlled studies aimed at understanding specific biological, chemical or physical effects. Furthermore, bioreactors allow for a safe and reproducible production of tissue constructs. For later clinical applications, the bioreactor system should be an advantageous method in terms of low contamination risk, ease of handling and scalability. To date the goals and expectations of bioreactor development have been fulfilled only to some extent, as bioreactor design in tissue engineering is very complex and still at an early stage of development. In this review we summarize important aspects for bioreactor design and provide an overview on existing concepts. The generation of three dimensional cartilage-carrier constructs is described to demonstrate how the properties of engineered tissues can be improved significantly by combining biological and engineering knowledge. In the future, a very intimate collaboration between engineers and biologists will lead to an increased fundamental understanding of complex issues that can have an impact on tissue formation in bioreactors. PMID- 16243272 TI - Directed evolution of bacterial alanine racemases with higher expression level. AB - Bacterial alanine racemase (EC 5.1.1.1) is a pyridoxal 5'-phosphate-dependent enzyme that catalyzes the interconversion of L-alanine and D-alanine. It can be classified into two groups: biosynthetic enzymes with low catalytic activity and catabolic enzymes with high catalytic activity. It can react with serine to a limited extent. Two biosynthetic alanine racemase genes in Escherichia coli and Salmonella typhimurium were DNA shuffled, and a very diverse chimeric gene library was constructed. An E. coli serine auxotroph was transformed with the shuffled genes, and the recombinant clones were screened on selective media supplemented with 0.5-5 mM D-serine as an L-serine supplier. Selected clones were expected to contain racemases exhibiting higher catalytic activities toward alanine as well as serine. Three independent clones that grew on selective media were isolated. The specific activities of crude extracts prepared from cells expressing the chimeric racemases were increased up to approximately three times more than those expressing the parental enzymes. The best chimera Ser15 racemase was expressed at a level approximately twofold higher than the parental alanine racemases. This high protein expression was demonstrated to be posttranscriptionally achieved. PMID- 16243273 TI - O-palmitoylcurdlan sulfate (OPCurS)-coated liposomes for oral drug delivery. AB - O-Palmitoylcurdlan sulfate (OPCurS) was applied to the liposomal surface to improve the stability of liposomes. To synthesize OPCurS, curdlan was chemically sulfated and then modified with a palmitoyl derivative. The synthesized OPCurS was characterized by Fourier transform-infrared spectroscopy (FT-IR). OPCurS coated liposomes prepared by the solvent evaporation method were characterized for size, shape, surface charge, and stability in simulated gastric fluid (SGF) and sodium cholate solution. The sizes of OPCurS-coated liposomes increased with the OPCurS content of liposomes and zeta potential decreased when OPCurS was applied to the liposomal surface. With the increase in the content of OPCurS attached to the liposomal surface, the stability of liposomes in SGF and sodium cholate solution was gradually induced and the stability was most improved at a lipid/OPCurS weight ratio of 1.5. Liposomes not coated with OPCurS released 99.5+/-2.3% of the initial 5-carboxyfluorescein (5-CF) content, whereas OPCurS coated liposomes released 53.7+/-3.7%. OPCurS on the surface of liposomes suppressed the release of 5-CF. Theses results indicate that OPCurS-coated liposomes can be effectively used as a drug delivery carrier via oral administration. PMID- 16243274 TI - Hydrogen and ethanol production from glycerol-containing wastes discharged after biodiesel manufacturing process. AB - H2 and ethanol production from glycerol-containing wastes discharged after a manufacturing process for biodiesel fuel (biodiesel wastes) using Enterobacter aerogenes HU-101 was evaluated. The biodiesel wastes should be diluted with a synthetic medium to increase the rate of glycerol utilization and the addition of yeast extract and tryptone to the synthetic medium accelerated the production of H2 and ethanol. The yields of H2 and ethanol decreased with an increase in the concentrations of biodiesel wastes and commercially available glycerol (pure glycerol). Furthermore, the rates of H2 and ethanol production from biodiesel wastes were much lower than those at the same concentration of pure glycerol, partially due to a high salt content in the wastes. In continuous culture with a packed-bed reactor using self-immobilized cells, the maximum rate of H2 production from pure glycerol was 80 mmol/l/h yielding ethanol at 0.8 mol/mol glycerol, while that from biodiesel wastes was only 30 mmol/l/h. However, using porous ceramics as a support material to fix cells in the reactor, the maximum H2 production rate from biodiesel wastes reached 63 mmol/l/h obtaining an ethanol yield of 0.85 mol/mol-glycerol. PMID- 16243275 TI - Dibenzothiophene desulfurizing enzymes from moderately thermophilic bacterium Bacillus subtilis WU-S2B: purification, characterization and overexpression. AB - The moderately thermophilic bacterium Bacillus subtilis WU-S2B desulfurized dibenzothiophene (DBT) at 50 degrees C through the selective cleavage of carbon sulfur bonds. In this study, three enzymes involved in the microbial DBT desulfurization were purified and characterized. The first two enzymes, DBT monooxygenase (BdsC) and DBT sulfone monooxygenase (BdsA), were purified from the wild-type strain, and the last one, 2'-hydroxybiphenyl 2-sulfinic acid desulfinase (BdsB), was purified from the recombinant Escherichia coli overexpressing the gene, bdsB, with chaperonin genes, groEL/ES. The genes of BdsC and BdsA were also overexpressed. The molecular weights of BdsC and BdsA were determined to be 200 and 174 kDa, respectively, by gel filtration chromatography, suggesting that both enzymes had four identical subunits. BdsB had a monomeric structure of 40 kDa. The three enzymes were characterized and compared with the corresponding enzymes (DszC, DszA, and DszB) of mesophilic desulfurization bacteria. The specific activities of BdsC, BdsA, and BdsB were 84.2, 855, and 280 units/mg, respectively, and the latter two activities were higher than those of DszA and DszB. The heat stability and optimum temperature of BdsC, BdsA, and BdsB were higher than those of DszC, DszA, and DszB. Other enzymatic properties were investigated in detail. PMID- 16243276 TI - Synthesis of linoleoyl disaccharides through lipase-catalyzed condensation and their surface activities. AB - Monolinoleoyl trehalose, maltose and cellobiose were synthesized by Candida antarctica lipase-catalyzed condensation in an organic solvent with a low water content. The use of a mixture of pyridine and tert-butanol as the reaction medium resulted in a high product concentration on the order of mmol/l for the synthesis of linoleoyl trehalose and maltose. The highest product concentration was achieved with the 0.4 volumetric fraction of pyridine. Linoleoyl cellobiose was also synthesized although its concentration was approximately one tenth the concentrations of linoleoyl trehalose and maltose. The surfactant properties of linoleoyl trehalose, maltose and cellobiose were measured. Among the esters, linoleoyl trehalose showed the strongest surface activity. PMID- 16243277 TI - Therapeutic use of phage cocktail for controlling Escherichia coli O157:H7 in gastrointestinal tract of mice. AB - To investigate the therapeutical use of phage mixture for controlling gastrointestinal Escherichia coli O157:H7 cells, in vitro and in vivo experiments were conducted. Three phages, SP15, SP21, and SP22 were selected from 26 phage stock screened from feces of stock animals and sewage influent. Addition of single or binary phage to the E. coli cell batch-culture reduced the turbidity of the culture. However, reascend of the turbidity due to the appearance of phage resistance cell was observed. On the other hand, addition of three phage mixture (SP15-21-22) did not produce reascend of culture turbidity under aerobic condition. Under anaerobic condition, slight reascend of culture turbidity was observed after SP15-21-22 addition. Chemostat continuous culture was operated under anaerobic condition to optimize the titer of phage cocktail and frequency of the addition for controlling E. coli cells. Five-log decrease of E. coli cell concentration after addition of phage cocktail of 10(9) Plaque forming unit (PFU)/ml was observed. However, reascend of cell concentration was observed after 1 d incubation. Repeated addition of phage cocktail was effective to reduce the cell concentration. Suspension of phage cocktail in the buffer containing 0.25% CaCO3 neutralized 9 times much more buffer of pH 2. Based on this in vitro experiment, phage cocktail (SP15-21-22) suspended in the buffer containing 0.25% CaCO3 was orally administrated to the mice in which E. coli O157:H7 cells was administrated in 2-d advance. E. coli and phage concentration in the feces was monitored for 9 d after phage addition. High titer of phage was detected in the feces when the phage cocktail administrated daily. E. coli O157:H7 concentration in the feces has been reduced according to the time period. However, difference of E. coli concentration in the feces of mice administrates with phage and in the control mice without phage addition became slight after 9-d test period. High titer of the phage settled down in the gastrointestinal tracts and reduced the concentration of E. coli cell. Repeated oral administration of SP15-21-22 was effective for rapid evacuation of E. coli O157:H7 from the feces and gastrointestinal tract of mice. PMID- 16243278 TI - A new microbial method for more efficient production of Nalpha-benzyloxycarbonyl L-aminoadipate delta-semialdehyde and Nalpha-benzyloxycarbonyl-D-aminoadipate delta-semialdehyde. AB - A new biochemical method for more efficient production of Nalpha benzyloxycarbonyl-L-aminoadipate delta-semialdehyde (Nalpha-Z-L-AASA) and Nalpha Z-D-AASA was developed with cells of Rhodococcus sp. AIU Z-35-1. Using the cells harvested after 1 d of cultivation, more than 95 mM Nalpha-Z-L-AASA was produced from 100 mM Nalpha-Z-L-lysine by incubating at pH 5.0 for 1 d at 30 degrees C or by incubating at pH 7.0 for 2 d at 10 degrees C. A similar conversion yield of Nalpha-Z-D-AASA was also obtained under the same conditions. These reaction times required were 1/4 and 1/2 of the respective ones by the method with amine oxidase, and the yields of Nalpha-Z-L-AASA and Nalpha-Z-D-AASA were 2 times higher than the respective ones by the method with amine oxidase. In addition, this method had the advantages of not requiring purification of enzyme and addition of catalase. Thus, the microbial method proposed here was superior to the chemical and other biochemical methods in simplicity, reaction rate, and yield. PMID- 16243279 TI - Cell microarray for screening feeder cells for differentiation of embryonic stem cells. AB - Microarrays are currently recognized as one of major tools in the assessment of gene expression via cDNA or RNA analysis and are now accepted as a powerful experimental tool for high-throughput screening of a large number of samples, such as cDNA and siRNAs. In this study, we examined the potential of the microarray methodology for high-throughput screening of candidate cells as feeder cells which effectively differentiate embryonic stem (ES) cells to the specific lineage. Cell arrays were prepared by applying three kinds of cells, PA6, human umbilical vein endothelial, and COS-1 cells, to circular spots, 2 mm in diameter, on a glass plate, followed by the application of mouse ES cells to the cell microarray. After 8 d in culture, TuJ1 (neuron-specific class III beta-tubulin) immunocytochemical staining clearly demonstrated that only PA6 cell spots had the capability to induce ES cells to neuronal differentiation. Although this is a model experiment, these findings clearly indicate that the cell microarray will become a powerful tool for high-throughput screening large numbers of candidate feeder cells for specific differentiation. PMID- 16243280 TI - Performance of a styrene-degrading biofilter inoculated with Pseudomonas sp. SR 5. AB - Styrene removal was studied for 3 months in a laboratory-scale biofilter packed with a mixed packing material of peat and ceramic at a ratio of 1 to 1 on a dry weight basis and inoculated with Pseudomonas sp. SR-5. More than 90% removal efficiency (RE) was attained at 1-140 g/m3/h styrene loads under nitrogen-source limitation. When RE decreased to 70% after 30 d with an increase in styrene load, readdition of SR-5 and washing of the filter packing material restored the RE to more than 90% by maintaining the population of SR-5 at 1-10% of the total cell number. The maximum elimination capacity (EC) by kinetic analysis was estimated to be 290 g/m3/h. High conversion of the removed styrene carbon to CO2, and significantly small production of cell mass from the removed carbon were confirmed. PMID- 16243281 TI - Effect of Bcl-2 overexpression on cell cycle and antibody productivity in chemostat cultures of myeloma NS0 cells. AB - Chemostat cultures of NS0 cell lines were carried out at dilution rates ranging from 0.8 d(-1) to 0.2 d(-1). Compared with the control, the viable cell density of the Bcl-2 cell line was approximately 10% higher at 0.8 d(-1) and increased to 55% when the dilution rate was reduced to 0.2 d(-1). As the dilution rate was reduced, the viability of the two cultures diverged reaching a difference of 43% at 0.2 d(-1). The specific growth rate of the control cells was the same as the dilution rate down to a value of 0.6 d(-1). By contrast, the specific growth rate of Bcl-2 cells was parallel to the dilution rate down to a value as low as 0.3 d( 1). For both NS0 cell lines, the G1 cell population decreased, while the S and G2/M cell populations increased as the dilution rate was reduced. The antibody titer of the control cells increased from 7 to 21 microg.ml(-1) as the dilution rate was reduced from 0.8 to 0.2 d(-1). With an initial increase from 2 to 15 microg.ml(-1) as the dilution rate was reduced from 0.8 to 0.4 d(-1), the antibody titer of the Bcl-2 cells remained constant as the dilution rate was further reduced to 0.2 d(-1). A good correlation between specific antibody production rate and the percentage of G2/M cells was observed. PMID- 16243282 TI - Determination of concentration and binding affinity of antibody fragments by use of surface plasmon resonance. AB - An assay method using a surface plasmon resonance (SPR) biosensor has been developed that allows quantitative measurement of the specific antibody concentration in crude materials. By injecting non-labeled antibody samples onto a biosensor surface on which antigen was immobilized at high densities, the concentration of active antibodies can be accurately measured. To clarify applicability of this method to pharmacokinetic studies, the concentration of active antibodies in mouse plasma was measured for 4 h after injection of antibodies in mice. Although this period of measurement might be insufficient for determining the pharmacokinetics of blood pool clearance, this method has some advantages over conventional methods in measurement of single-chain antibody fragment (scFv) concentrations. Using the SPR biosensor, scFv and antibodies without epitope tag peptides were easily detected in real time, requiring as little as 20 mul of blood sample. Moreover, from the apparent dissociation rate in the dissociation phase of the sensorgrams, we could identify whether the antibody fragments existed as bivalent or monovalent in animal blood. We also evaluated the antigen binding activity of the scFvs against human CD47 and found scFvs had slightly weak affinity to their antigen (K(D), about 10 nM) compared with F(ab')2 and Fab' fragments (K(D), about 3-4 nM). This assay method promises to be a convenient tool for quality control, screening, and simple pharmacokinetic analysis of antibody fragments and other recombinant proteins not having epitope tags. PMID- 16243283 TI - Purification and characterization of the alcohol dehydrogenase with a broad substrate specificity originated from 2-phenylethanol-assimilating Brevibacterium sp. KU 1309. AB - A novel 2-phenylethanol dehydrogenase has been purified from a soil bacterium Brevibacterium sp. KU 1309. The enzyme was purified about 1400-fold to homogeneity, and found to be a monomeric enzyme of apparent 39 kDa. The enzyme had broad substrate specificity and catalyzes a reversible oxidation of various primary alcohols to aldehydes. The enzyme required NAD+, but not NADP+ as a cofactor. Thus, the enzyme was classified into a group of NAD+-dependent primary alcohol dehydrogenase. The activity was inhibited by Cu2+, Ni2+, Ba2+, Hg2+ and p chloromercuribenzoate. The enzyme is expected to be applicable as an effective biocatalyst in the oxidation of various alcohols. PMID- 16243284 TI - Active lactonizing lipase (LipL) efficiently overproduced by Pseudomonas strains as heterologous expression hosts. AB - Pseudomonas sp. strain 109 secretes lactonizing lipase (LipL), which catalyzes efficient intramolecular transesterification of omega-hydroxyfatty acid esters to form macrocyclic lactones. Because Escherichia coli was found to be unsuitable as an expression host due to the predominant formation of inactive LipL-inclusion bodies and a lack of proper secretion machinery which is also required for the formation of active LipL, Pseudomonas strains were surveyed as expression hosts. Pseudomonas sp. strain 109, an original LipL producer, showed a 7.1-fold higher level of active LipL when the lipL gene under the control of tac-lacUV5 tandem promoter was introduced together with a limL gene encoding a LipL-specific chaperon. Pseudomonas aeruginosa ADD 1976 containing a T7 RNA polymerase gene in the chromosome and plasmid-borne lipL-limL genes under the control of T7 promoter showed a 13-fold higher level of active LipL. Several combinations in the number of lipL and/or limL genes on the plasmid were investigated, and (lipL)3-limL was found to be most efficient, yielding a 67-fold greater production of active LipL than that obtained by the wild-type Pseudomonas sp. strain 109. PMID- 16243285 TI - Patterning cultured cells by visible light illumination with photosensitizers. AB - We showed that PC12 cells and 3T3 cells cultured in dishes were killed by illumination with visible white light from a halogen lamp at 7 x 10(4) lx for 5 min in the presence of either 10 microM hematoporphyrin or 2 microM methylene blue as a photosensitizer. This simple technique, based on the photodynamic reaction via generation of reactive oxygen species can be applicable for patterning cultured cells. PMID- 16243286 TI - Production of L-talitol from L-psicose by Metschnikowia koreensis LA1 isolated from soy sauce mash. AB - A strain LA1 that can convert L-psicose to L-talitol was isolated from soy sauce mash and identified as Metschnikowia koreensis. The cells grown on L-arabitol were found to have relatively high conversion potential. Addition of D-sorbitol to the reaction mixture considerably accelerated the conversion rate of L-psicose to L-talitol. During the conversion reaction, D-sorbitol was added to the reaction mixture at 12-h intervals to maintain the concentration of D-sorbitol at 1.0%. The final conversion ratios were 81.4%, 75.2%, 73.0%, 60.4% and 43.5% using washed cells when the concentrations of L-psicose were 0.5%, 1.0%, 2.0%, 3.0% and 5.0%, respectively. The product from L-psicose was identified as L-talitol by HPLC analysis, and infrared spectroscopy, optical rotation and melting point measurements. PMID- 16243287 TI - Fed-batch culture under illumination with blue light emitting diodes (LEDs) for astaxanthin production by Haematococcus pluvialis. AB - To increase the cell concentration and the accumulation of astaxanthin, the effects of the fed-batch addition of 10-fold-concentrated medium to supply nutrients, as well as illumination with blue light emitting diodes (LEDs), on cell growth and accumulation of astaxanthin were studied for the cultivation of Haematococcus pluvialis. Using the fed-batch addition method, the cell concentration increased above 1 mg-dry cell/cm3, and under illumination with blue LEDs, the astaxanthin concentration reached approximately 70 microg/cm3. This method was much simpler to operate than the medium replacement method in operation and enabled us to attain a higher total yield of astaxanthin. PMID- 16243288 TI - Hyper expression of kojibiose phosphorylase gene and trehalose phosphorylase gene from Thermoanaerobacter brockii ATCC35047 in Bacillus subtilis and selaginose synthesis utilizing two phosphorylases. AB - The kojibiose phosphorylase (KP) gene and trehalose phosphorylase (TP) gene from Thermoanaerobacter brockii ATCC35047 were intracellularly hyper-expressed under the control of the Bacillus amyloliquefaciens alpha-amylase promoter in Bacillus subtilis. The production yields were estimated to be 2.1 g of KP and 4.9 g of TP per liter of medium. Selaginose, non-reducing trisaccharide, was synthesized from trehalose utilizing the recombinant KP and TP from B. subtilis. Selaginose was not hydrolyzed by salivary amylase, artificial gastric juice, pancreatic amylase, or small intestinal enzymes. PMID- 16243289 TI - High-density fluorescently labeled rolling-circle amplicons for DNA diagnostics. PMID- 16243290 TI - Spectrophotometric determination of hydroxylamine and nitrite in mixture in water and biological samples after micelle-mediated extraction. PMID- 16243291 TI - Adverse psychological outcomes in colorectal cancer screening: does health anxiety play a role? AB - People who are anxious about their health are more likely to misinterpret health information as personally threatening and less likely to be reassured by medical investigations that show they are free from disease. Consequently, health anxious people would be expected to react more adversely to cancer screening, but this possibility has rarely been explored. The moderating role of health anxiety on the psychological impact of participating in colorectal cancer screening was examined among a sub-sample of 3535 participants in a large, community-based trial of colorectal cancer screening in the UK. The screening modality was flexible sigmoidoscopy, which examines the bowel for pre-cancerous polyps. It was predicted that health anxiety would be associated with more worry about cancer before screening, a greater increase in worry if polyps were detected, and less reassurance after a clear result. As expected, health anxious participants were more anxious and more worried about bowel cancer both before and after screening. However, they experienced greater reductions in anxiety and worry about cancer following the examination. They reported lower levels of reassurance following screening, but also expressed more positive reactions to the experience. The positive psychological benefits of attending medical investigations should be examined in future work, because this may go some way towards explaining why health anxious people repeatedly seek medical interventions. PMID- 16243292 TI - Fibrocystin interacts with CAML, a protein involved in Ca2+ signaling. AB - The predicted structure of the autosomal recessive polycystic kidney disease protein, fibrocystin, suggests that it may function as a receptor, but its function remains unknown. To understand its function, we searched for proteins that interact with the intracellular C-terminus of fibrocystin using the yeast two-hybrid system. From the screening, we found calcium modulating cyclophilin ligand (CAML), a protein involved in Ca(2+) signaling. Immunofluorescent analysis showed that both proteins are co-localized in the apical membrane, primary cilia, and the basal body of cells derived from the distal nephron Epitope-tagged expression constructs of both proteins were co-immunoprecipitated from COS7 cells. The intracellular C-terminus of fibrocystin interacts with CAML, a protein with an intracellular distribution that is similar to that of PKD2. Fibrocystin may participate in regulation of intracellular Ca(2+) in the distal nephron in a manner similar to PKD1 and PKD2 that are involved in autosomal dominant polycystic kidney disease. PMID- 16243293 TI - The ABC transporter PGP-2 from Caenorhabditis elegans is expressed in the sensory neuron pair AWA and contributes to lysosome formation and lipid storage within the intestine. AB - The functional role of the ABC transporter PGP-2 from the nematode Caenorhabditis elegans has been studied by combining phenotype analyses of pgp-2 deletion mutants or pgp-2 RNAi treated worms with reporter gene studies using a pgp-2::GFP construct. pgp-2 mutants showed a strong reduction of lipid stores. In addition, we found that in the case of the pgp-2 mutant or after pgp-2 RNAi the worms were unable to perform pinocytosis and to acidify intestinal lysosomes. Especially under cholesterol-restricted conditions, the viability of the mutant was reduced. Surprisingly, the chemosensory AWA neurons in the head region were identified as expression sites by reporter gene studies. These neurons are known to be involved in attraction behaviour towards odorants associated with potential food bacteria. Our results imply that PGP-2 is involved in a signalling process that connects sensory inputs to intestinal functions, possibly by influencing acidification of intestinal lysosomes, which in turn may affect pinocytosis and lipid storage. PMID- 16243294 TI - Permeable ions differentially affect gating kinetics and unitary conductance of L type calcium channels. AB - Although ion permeation and gating of L-type Ca(2+) channels are generally considered separate processes controlled by distinct components of the channel protein, ion selectivity can vary with the kinetic state. To test this possibility, we studied single-channel currents (cell-attached) of recombinant L type channels (Ca(V)1.2, beta(2a), and alpha(2)delta) transiently expressed in tsA201 cells in the presence of the channel agonist BayK 8644 which promotes long channel openings (Mode 2 openings). We found that both the brief (Mode 1) and long (Mode 2) mean open times in the presence of Ca(2+) were relatively longer than those with Ba(2+). The unitary slope conductance with Ba(2+) was significantly larger (p<0.05) in Mode 2 openings than for brief Mode 1 openings, whereas the conductance with Ca(2+) did not vary with mode gating. Consequently, the gamma(Ba):gamma(Ca) ratio was greater for Mode 2 than Mode 1 openings. Our findings indicate that both ion permeation and gating kinetics of the L-type channel are differentially modulated by permeable ions. Ca(2+) binding to the L type channel may stabilize the alteration of channel ion permeability mediated by gating kinetics, and thus, play a role in preventing excessive ion entry when the activation gating of the channel is promoted to the prolonged open state. PMID- 16243295 TI - Differentiation-inducing factor-1 suppresses gene expression of cyclin D1 in tumor cells. AB - To determine the mechanism by which differentiation-inducing factor-1 (DIF-1), a morphogen of Dictyostelium discoideum, inhibits tumor cell proliferation, we examined the effect of DIF-1 on the gene expression of cyclin D1. DIF-1 strongly reduced the expression of cyclin D1 mRNA and correspondingly decreased the amount of beta-catenin in HeLa cells and squamous cell carcinoma cells. DIF-1 activated glycogen synthase kinase-3beta (GSK-3beta) and inhibition of GSK-3beta attenuated the DIF-1-induced beta-catenin degradation, indicating the involvement of GSK 3beta in this effect. Moreover, DIF-1 reduced the activities of T-cell factor (TCF)/lymphoid enhancer factor (LEF) reporter plasmid and a reporter gene driven by the human cyclin D1 promoter. Eliminating the TCF/LEF consensus site from the cyclin D1 promoter diminished the effect of DIF-1. These results suggest that DIF 1 inhibits Wnt/beta-catenin signaling, resulting in the suppression of cyclin D1 promoter activity. PMID- 16243296 TI - Osteoclasts but not osteoblasts are affected by a calcified surface treated with zoledronic acid in vitro. AB - Bisphosphonates are potent inhibitors of osteoclast-mediated bone resorption. Recent interest has centered on the effects of bisphosphonates on osteoblasts. Chronic dosing of osteoblasts with solubilized bisphosphonates has been reported to enhance osteogenesis and mineralization in vitro. However, this methodology poorly reflects the in vivo situation, where free bisphosphonate becomes rapidly bound to mineralized bone surfaces. To establish a more clinically relevant cell culture model, we cultured bone cells on calcium phosphate coated quartz discs pre-treated with the potent nitrogen-containing bisphosphonate, zoledronic acid (ZA). Binding studies utilizing [(14)C]-labeled ZA confirmed that the bisphosphonate bound in a concentration-dependent manner over the 1-50microM dose range. When grown on ZA-treated discs, the viability of bone-marrow derived osteoclasts was greatly reduced, while the viability and mineralization of the osteoblastic MC3T3-E1 cell line were largely unaffected. This suggests that only bone resorbing cells are affected by bound bisphosphonate. However, this system does not account for transient exposure to unbound bisphosphonate in the hours following a clinical dosing. To model this event, we transiently treated osteoblasts with ZA in the absence of a calcified surface. Osteoblasts proved highly resistant to all transitory treatment regimes, even when utilizing ZA concentrations that prevented mineralization and/or induced cell death when dosed chronically. This study represents a pharmacologically more relevant approach to modeling bisphosphonate treatment on cultured bone cells and implies that bisphosphonate therapies may not directly affect osteoblasts at bone surfaces. PMID- 16243297 TI - A functional link between Disrupted-In-Schizophrenia 1 and the eukaryotic translation initiation factor 3. AB - Disrupted-In-Schizophrenia 1 (DISC1) was identified as a candidate gene for schizophrenia. DISC1 is disrupted by a balanced t(1;11)(q42.1;q14.3) translocation segregating with schizophrenia and related psychiatric illness in a large Scottish family. Here, we show that DISC1 interacts via its globular domain with the p40 subunit of the eukaryotic translation initiation factor 3. Furthermore, we found that overexpression of DISC1 in SH-SY5Y cells induces the assembly of eIF3- and TIA-1-positive stress granules (SGs), discrete cytoplasmic granules formed in response to environmental stresses. Our findings suggest that DISC1 may function as a translational regulator and may be involved in stress response. PMID- 16243298 TI - Reduced transcriptional activity in individuals with IL-18 gene variants detected from functional but not association study. AB - Genetic polymorphisms of IL-18 and its receptor were reported to be associated with elevated serum IgE levels, atopy, and/or asthma. However, conflicting results were observed in various association studies and functional activity of these polymorphisms remains unclear. A total of 393 unrelated subjects were involved in this study. Direct PCR-sequencing method was used to screen novel polymorphisms. The functional significance of these polymorphisms was investigated using reporter gene assay. Three known (-137, +113, and +127) polymorphisms in the IL-18 promoter were identified with a perfect linkage disequilibrium (Delta=1, p<0.001) among them. No significant difference in the genotype frequencies of these polymorphisms between atopy and atopic phenotypes in Singaporean Chinese, Malays, and Indians was observed. However, transcriptional activities were significantly increased in HepG2 cultured cells with wild-type IL-18 genotype (-137/G, +113/T, and +127/C) than mutated genotype (-137/C, +113/G, and +127/T). Although these polymorphisms appear to have no association with atopic phenotypes in our population, subsequent functional studies suggest that polymorphisms in the IL-18 promoter region could affect significantly its activity. PMID- 16243299 TI - Extracellular release of BACE1 holoproteins from human neuronal cells. AB - BACE1 is a membrane-bound aspartyl protease involved in production of the Alzheimer's amyloid beta-protein. The BACE1 ectodomain is partially cleaved to generate soluble BACE1, but the physiological significance of this event is unclear. During our characterization of BACE1 shedding from human neuroblastoma SH-SY5Y cells stably expressing BACE1, we unexpectedly found that detectable amounts of BACE1 holoproteins were released extracellularly along with soluble BACE1. Treatment with the metalloprotease inhibitor, TAPI-1, inhibited BACE1 shedding but increased BACE1 holoprotein release. Soluble and full-length BACE1 were released in parallel, at least partly originating from the plasma membrane. Furthermore, the release of soluble BACE1, but not full-length BACE1, was increased by deletion of the C-terminal dileucine motif, indicating that dysregulated BACE1 sorting affects BACE1 shedding. These findings suggest that the release of BACE1 holoproteins may be a physiologically relevant cellular process. PMID- 16243300 TI - The expression of CCL2 by T lymphocytes of mammary tumor bearers: role of tumor derived factors. AB - Tumor-associated chemokines, including CC chemokine ligand 2/monocyte chemoattractant protein-1 (CCL2), are thought to play many roles in cancer progression. Here we demonstrate the novel finding that during growth of the D1 7,12-dimethylbenzanthracene-3 mammary tumor in BALB/c mice, there is a dramatic up-regulation of CCL2 in splenic T cells at both the mRNA and protein levels upon stimulation. Of particular relevance is the finding that tumor-infiltrating T cells also produce high levels of CCL2. While a variety of tumor cell lines have been found to produce CCL2, we found no detectable levels of CCL2 protein in supernatants of the cultured mammary tumor cells. Investigation of the mechanisms involved in CCL2 induction showed that treatment of splenic T cells with the tumor-derived factors GM-CSF and phosphatidyl serine (PS) resulted in increased CCL2 production. This increased production may be involved in the downregulation of IFN-gamma by the T cells of tumor-bearing mice previously reported in this model, as treatment of splenic T lymphocytes with CCL2 resulted in a decreased secretion of IFN-gamma by those cells. PMID- 16243301 TI - Peripheral nerve esterases and the promotion of organophosphate-induced neuropathy in hens. AB - Several esterase inhibitors, not capable of causing peripheral neuropathy by themselves, exacerbate organophosphate-induced delayed polyneuropathy (OPIDP) and other axonopathies. This effect was called promotion of axonopathies and it was found not to be associated with inhibition of neuropathy target esterase (NTE), the molecular target of OPIDP. The search for an esterase as the target of promotion has started long ago, when an eterogeneous group of esterases hydrolysing phenyl valerate (PV) was identified in hen's sciatic nerve by means of selective inhibitors. Correlation studies in vivo indicated that the target of promotion may have been among the proteins present in the soluble fraction. When this soluble PV-esterase activity was separated on a Sephacryl-S-300 column, correlation was found between promotion and its inhibition in vivo. The electrophoretic analysis of this fraction indicated the presence of several proteins. Subsequent ion-exchange chromatography identified a protein of about 80 kDa molecular weight that was associated with PV-esterase activity. The inhibition of this activity did also correlate with promotion. The sequence of this protein identified it as ovotransferrin, but commercial preparations of ovotransferrin were found to lack PV-esterase activity. Binding experiments on this purified PV-activity and on commercial ovotransferrin using radiolabelled promoters were inconclusive. Titration of this PV-activity showed that about 20 30% of it is resistant to high concentrations of several inhibitors, suggesting heterogeneity of the fraction. In fact, bi-dimensional electrophoresis indicated the presence of several proteins. Finally, in vivo correlation experiments with p toluensulfonyl fluoride showed that whereas this chemical does not promote OPIDP induced by dibutyl dichlorovinyl phosphate, it does inhibit about 80% of this PV activity. In conclusion, available data indicate that the target of promotion is unlikely to be ovotransferrin. However, all promoters identified so far are esterase inhibitors suggesting that the target of promotion might be, indeed, a protein with esteratic activity. PMID- 16243302 TI - Visualizing viral transduction of a cocaine-hydrolyzing, human butyrylcholinesterase in rats. AB - Human plasma butyrylcholinesterase (BChE) is essential for cocaine detoxification even though its catalytic efficiency for that substrate is relatively poor. Site directed mutagenesis of this protein has recently been used to obtain much improved cocaine esterases, one of which we designate CocE. We previously showed that adenoviral transduction of such esterases caused up to 50,000-fold increases in circulating cocaine hydrolase activity, led to drastically shortened cocaine half-life, and blunted the cardiovascular responses to cocaine in rats. In those experiments, gene transduction of cocaine esterase was sustained at high levels for up to 1 week but then declined steeply. Our eventual goal is to use long-term esterase expression as a means of reducing drug reward and extinguishing intake in models of cocaine-addiction. Therefore, we investigated the site of enzyme transduction for clues to the local reactions that may limit the duration of CocE expression. Histological and immunohistochemical observations demonstrated that hepatocytes were the primary focus for transduction of modified human BChE. Rats were administered 2.2 x 10(10) plaque forming units of a replication-incompetent, type-5 adenoviral vector incorporating CocE cDNA. Within days the livers showed intense thiocholine staining for BChE activity. Selective immunohistochemistry for human BChE proved that this activity represented CocE transgene. By 5 days, however, pockets of mononuclear cells had invaded the hepatic parenchyma, and a meshwork of IgM-like immunoreactivity had lined the hepatic sinusoids. These phenomena probably represent early responses of the immune system, either to the transduced CocE or to the hepatocytes producing this protein. PMID- 16243303 TI - Nematode acetylcholinesterases are encoded by multiple genes and perform non overlapping functions. AB - Nematodes are unusual in that diverse molecular forms of acetylcholinesterase are the product of distinct genes. This is best characterised in the free living organism Caenorhabditis elegans, in which 3 genes are known to give rise to distinct enzymes, with a fourth likely to be non-functional. ACE-1 is an amphiphilic tetramer associated with a hydrophobic non-catalytic subunit, analogous to vertebrate T enzymes, whereas ACE-2 and ACE-3 are glycosylphosphatidylinositol-linked amphiphilic dimers. The different ace genes show distinct anatomical patterns of expression in muscles, sensory neurons and motor neurons, with only a few examples of coordinated expression. Clear homologues of ace-1 and ace-2 have now been isolated from a variety of parasitic nematodes, and the predicted proteins have very similar C-terminal amino acid sequences, implying an analogous means of anchorage to membranes. In addition to these membrane-bound enzymes, many parasitic nematodes which colonise mucosal surfaces secrete acetylcholinesterases to the external (host) environment. These hydrophilic enzymes are separately encoded in the genome, so that some parasites may thus have a total complement of six ace genes. The secretory enzymes have been characterised from the intestinal nematode Nippostrongylus brasiliensis and the lungworm Dictyocaulus viviparus. These show a number of common features, including a truncated C-terminus and an insertion at the molecular surface, when compared to other nematode acetylcholinesterases. Although the function of these enzymes has not been determined, they most likely alter host physiological responses to promote survival of the parasite. PMID- 16243304 TI - Serine hydrolase targets of organophosphorus toxicants. AB - Acetylcholinesterase (AChE) is one of several hundred serine hydrolases in people potentially exposed to about 80 organophosphorus (OP) compounds important as insecticides or chemical warfare agents. The toxicology of OPs was interpreted until recently almost solely on the basis of AChE inhibition. It is assumed that each serine hydrolase has a specific function and proposed that every OP compound has a unique inhibitory profile. This review considers the progress in sifting the expanding list of potential serine hydrolase toxicological targets. About 50 serine hydrolase targets have been recognized but only a few studied thoroughly. The toxicological relevance of known secondary OP targets is established mainly from observations with humans (butyrylcholinesterase and neuropathy target esterase-lysophospholipase) and studies with mice (cannabinoid CB1 receptor, carboxylesterase, lysophospholipase and platelet activating factor acetylhydrolase) and hen eggs (arylformamidase or kynurenine formamidase). Pesticides most commonly shown to inhibit these targets in experimental vertebrates are chlorpyrifos and tribufos. Generally the levels of environmental and occupational OP pesticide exposure are well below those causing in vivo inhibition of secondary serine hydrolase targets. Although exposure to OP insecticides is decreasing from stricter regulations and the development of resistant pest strains, it will continue to some degree for decades in the future. Only two OPs are used as pharmaceuticals, i.e. echothiophate as an ophthalmic for treatment of glaucoma and metrifonate as an anthelmintic for Schistosoma (and formerly as a candidate drug for improved cognitive function in Alzheimer's disease). In safety evaluations, knowledge on known OP targets must be balanced against major gaps in current understanding since more than 75% of the serine hydrolases are essentially unknown as to OP targeting and relevance, i.e. it is not clear if they play a role in OP toxicology. PMID- 16243305 TI - Peripheral cholinergic disturbances in Alzheimer's disease. AB - The most pronounced neurochemical abnormality in Alzheimer's disease (AD) is cholinergic dysfunction in the central nervous system. Peripheral tissues may also be affected, however, including blood. The present study undertook to determine the activity of acetylcholinesterase (AChE) and its molecular forms in erythrocytes, lymphocytes and platelets of normal elderly subjects and probable AD cases. These samples contained dimeric globular (G2), tetrameric globular (G4) and asymmetric (A12) AChE forms, but no globular monomeric (G1) enzyme. In both lymphocytes and platelets, the major AChE molecular form was G2 (approximately 80%), with G4 and A12 forms accounting for nearly equal portions of the remainder. Total AChE activities and measured sedimentation coefficients were similar in the control and AD samples (from patients with mild and moderately severe cognitive deficiency). However, the groups differed significantly in the proportion of certain AChE molecular forms. Thus, as compared with controls, the amount of A12 AChE in the AD samples was increased 148 and 161% in lymphocytes and platelets, respectively. Genotyping for apolipoprotein E (ApoE) and the butyrylcholinesterase K (BCHE-K) variant, carried out using the polymerase chain reaction, showed that AD patients carried the ApoE4 allele at a significantly higher frequency than the controls. On the other hand there were no significant group differences in the occurrence of the BCHE-K variant and no synergism between ApoE alleles and the BCHE-K variant in our Hungarian AD population. PMID- 16243306 TI - Delivery of human acetylcholinesterase by adeno-associated virus to the acetylcholinesterase knockout mouse. AB - The purpose of this work was to develop a gene delivery system that expressed acetylcholinesterase (AChE) for prolonged periods. An adeno-associated virus (AAV) expressing human AChE was constructed by co-transfecting three plasmids into HEK 293T cells. The purified vector expressed 0.17 microg AChE per 1 million viral particles in culture medium in 23 h, or 0.8 U/ml. The AAV/hAChE was injected into muscle of adult AChE knockout mice and into the brains of 3-6 week old AChE knockout mice. Intramuscular injection yielded plasma AChE levels approaching 50% of the AChE activity of wild-type mouse plasma. The highest AChE activity was found on day 3 post-injection. AChE activity declined thereafter to a constant 7% of normal. The decreased level was accompanied by the appearance of anti-human AChE antibodies, suggesting partial clearance of AChE from plasma by antibodies. Intrastriatal injection resulted in AChE expression in the striatum. No antibodies were detected in animals treated intrastriatally. Motor coordination was improved and the lifespan of intrastriatally-treated AChE knockout mice was prolonged. Human AChE was expressed in mouse brain for up to 7 months after intrastriatal injection of an AAV/hAChE construct. Gene-therapy to supply AChE to the striatum improved motor coordination and prolonged the life of mice genetically deficient in AChE, probably by reducing their susceptibility to spontaneous seizures. This supports the hypothesis that their seizures are induced by excess acetylcholine. PMID- 16243307 TI - Quantification of succinylacetone in urine of hepatorenal tyrosinemia patients by HPLC with fluorescence detection. AB - BACKGROUND: Hepatorenal tyrosinemia (HT1) is considered a treatable inherited metabolic disease, particularly when detected early in life. Succinylacetone (SA), a unique metabolic marker for HT1, is normally circulating or excreted at very low physiological concentrations and is significantly increased in HT1 patients. METHODS: We developed and validated a new method for the determination of SA in urine using high-pressure liquid chromatography with fluorescence detection. SA and its homologue 5,7-dioxooctanoic acid used as internal standard (IS) were extracted from urine, derivatized with pyrenebutyric hydrazide and separated on a C18 column within 11 min. Calibration curves were linear between 0.025 to 100 micromol/l. Within- and between-day variations were <5% and results obtained by the current method compared favorably with a reference liquid chromatography tandem mass spectrometric method. The method was applied retrospectively to the analysis of urine samples from HT1 patients. CONCLUSIONS: The method requires a minimal sample volume (0.1 ml) with simple instrumentation. The method enabled us to differentiate HT1 cases (n=14) from controls (n=104), regardless of the years of urine storage. PMID- 16243308 TI - Characterization of innexin gene expression and functional roles of gap junctional communication in planarian regeneration. AB - Planaria possess remarkable powers of regeneration. After bisection, one blastema regenerates a head, while the other forms a tail. The ability of previously adjacent cells to adopt radically different fates could be due to long-range signaling allowing determination of position relative to, and the identity of, remaining tissue. However, this process is not understood at the molecular level. Following the hypothesis that gap-junctional communication (GJC) may underlie this signaling, we cloned and characterized the expression of the Innexin gene family during planarian regeneration. Planarian innexins fall into 3 groups according to both sequence and expression. The concordance between expression based and phylogenetic grouping suggests diversification of 3 ancestral innexin genes into the large family of planarian innexins. Innexin expression was detected throughout the animal, as well as specifically in regeneration blastemas, consistent with a role in long-range signaling relevant to specification of blastema positional identity. Exposure to a GJC-blocking reagent which does not distinguish among gap junctions composed of different Innexin proteins (is not subject to compensation or redundancy) often resulted in bipolar (2-headed) animals. Taken together, the expression data and the respecification of the posterior blastema to an anteriorized fate by GJC loss-of-function suggest that innexin-based GJC mediates instructive signaling during regeneration. PMID- 16243309 TI - Bmp signaling promotes intermediate mesoderm gene expression in a dose-dependent, cell-autonomous and translation-dependent manner. AB - The intermediate mesoderm lies between the somites and the lateral plate and is the source of all kidney tissue in the developing vertebrate embryo. While bone morphogenetic protein (Bmp) signaling is known to regulate mesodermal cell type determination along the medio-lateral axis, its role in intermediate mesoderm formation has not been well characterized. The current study finds that low and high levels of Bmp ligand are both necessary and sufficient to activate intermediate and lateral mesodermal gene expression, respectively, both in vivo and in vitro. Dose-dependent activation of intermediate and lateral mesodermal genes by Bmp signaling is cell-autonomous, as demonstrated by electroporation of the avian embryo with constitutively active Bmp receptors driven by promoters of varying strengths. In explant cultures, Bmp activation of Odd-skipped related 1 (Odd-1), the earliest known gene expressed in the intermediate mesoderm, is blocked by cyclohexamide, indicating that the activation of Odd-1 by Bmp signaling is translation-dependent. The data from this study are integrated with that of other studies to generate a model for the role of Bmp signaling in trunk mesodermal patterning in which low levels of Bmp activate intermediate mesoderm gene expression by inhibition of repressors present in medial mesoderm, whereas high levels of Bmp repress both medial and intermediate mesoderm gene expression and activate lateral plate genes. PMID- 16243310 TI - Inhibitory action of L-type Ca2+ current by paeoniflorin, a major constituent of peony root, in NG108-15 neuronal cells. AB - The effects of paeoniflorin, a glycoside isolated from the root of Paeonia lactiflora, on ion currents in a mouse neuroblastoma and rat glioma hybrid cell line, NG108-15 were investigated. Paeoniflorin (1-300 microM) reversibly produced an inhibition of L-type voltage-dependent Ca2+ current (I(Ca,L)) in a concentration-dependent manner. Paeoniflorin caused no change in the overall shape of the current-voltage relationship of I(Ca,L). The IC50 value of paeoniflorin-induced inhibition of I(Ca,L) was 14 microM. However, neither adenosine deaminase (1 U/ml) nor 8-cyclopentyl-1, 3-dipropylxanthine (10 microM) could reverse the inhibition by paeoniflorin of I(Ca,L). Paeoniflorin (30 microM) shifted the steady-state inactivation curve of I(Ca,L) to more negative membrane potentials by approximately -10 mV. It also prolonged the recovery of I(Ca,L). The inhibitory effect of paeoniflorin on I(Ca,L) exhibited tonic and use dependent characteristics. Paeoniflorin could effectively suppress I(Ca,L) evoked by action potential waveforms. Paeoniflorin at a concentration of 30 microM produce a slight inhibition of voltage-dependent Na+ current and delayed rectifier K+ current. Under current-clamp configuration, unlike adenosine, this compound decreased the firing of action potentials. Taken together, this study indicates that paeoniflorin can block L-type Ca2+ channels in NG108-15 cells in a mechanism unlinked to the binding to adenosine receptors. The effects of paeoniflorin on ion currents may partly, if not entirely, contribute to the underlying mechanisms through which it affects neuronal or neuroendocrine function. PMID- 16243311 TI - Enhanced response of pig coronary arteries to endothelin-1 after ischemia reperfusion. Role of endothelin receptors, nitric oxide and prostanoids. AB - To analyse the coronary effects of endothelin-1 after ischemia-reperfusion, the left anterior descending coronary artery of anesthetized pigs was subjected to 30 min occlusion followed by 60-min reperfusion. Then, rings distal (ischemic arteries) and proximal (control arteries) to the occlusion were taken from this artery and prepared for isometric tension recording. The sensitivity of the contraction in response to endothelin-1 (3 x 10(-10)-3 x 10(-7) M) and the endothelin ET(B) receptor agonist IRL-1620 (3 x 10(-10)-3 x 10(-7) M) was greater in ischemic vessels. The endothelin ET(A) receptor antagonist BQ-123 (10(-7)-3 x 10(-6) M) decreased the sensitivity of the response to endothelin-1 similarly in ischemic and control arteries. The endothelin ET(B) receptor antagonist BQ-788 (10(-6) M), endothelium removal or the inhibitor of nitric oxide synthesis N(omega)-nitro-L-arginine methyl ester (L-NAME 10(-4) M) potentiated the response to endothelin-1 and IRL-1620 in control arteries only. The cyclooxygenase inhibitor meclofenamate (10(-5) M) augmented the maximal response to endothelin-1 in control arteries, and reduced it in ischemic arteries. In precontracted arteries, IRL-1620 (3 x 10(-11)-3 x 10(-10) M) relaxed control but not ischemic arteries, and L-NAME or meclofenamate abolished this relaxation. Therefore, ischemia-reperfusion increases the coronary vasoconstriction in response to endothelin-1 probably due to impairment of endothelin ET(B) receptor-induced release of nitric oxide and prostacyclin, augmentation of the contractile response to activation of endothelin ET(B) receptors, and increased release of vasoconstrictor prostanoids. PMID- 16243312 TI - Caspase 8/10 are not mediating apoptosis in neuroblastoma cells treated with CDK inhibitory drugs. AB - Olomoucine and Roscovitine are pharmacological inhibitors of cyclin-dependent kinases (CDK) displaying a promising profile as anticancer agents. Both compounds are effective inductors of apoptosis in a human neuroblastoma cell line, SH-SY5Y. The characterization of this process had suggested the involvement of an extrinsic pathway [Ribas, J., Boix, J., 2004. Cell differentiation, Caspase inhibition, and macromolecular synthesis blockage, but not Bcl-2 or Bcl-XL proteins, protect SH-SY5Y cells from apoptosis triggered by two CDK inhibitory drugs. Exp. Cell Res. 295 9-24.], which depends on either Caspase 8 or Caspase 10 activation. However, neither Caspase 8 nor Caspase 10 is expressed in SH-SY5Y cells because of gene silencing. Upon Olomoucine or Roscovitine treatment, no re expression of Caspase 8 or Caspase 10 was found. Therefore, in SH-SY5Y cells, this type of drugs is not triggering a canonical, Caspase 8/10-mediated, extrinsic apoptotic pathway. PMID- 16243313 TI - The "resurrection method" for modification of specific proteins in higher plants. AB - We describe a new method designated "the resurrection method" by which a modified protein is expressed in higher plants in place of the original protein. The modified gene constructed by introducing synonymous codon substitutions throughout the original gene to prevent the sequence-specific degradation of its mRNA during RNA silencing is expressed while the expression of the original gene is suppressed. Here, we report the successful alteration of the biochemical properties of green fluorescent protein expressed in transgenic Nicotiana benthamiana, suggesting that this method could be useful for gene control in living plants. PMID- 16243314 TI - SNARE-mediated membrane traffic modulates RhoA-regulated focal adhesion formation. AB - In the present study, we examined the role of soluble NSF attachment protein receptor (SNARE)-mediated membrane traffic in the formation of focal adhesions during cell spreading. CHO-K1 cells expressing a dominant-negative form of N ethylmaleimide-sensitive factor (E329Q-NSF) were unable to spread as well as control cells and they formed focal adhesions (FAs) that were larger than those in control cells. FA formation was impaired in cells transfected with a dominant negative form of RhoA, but, significantly, not in cells simultaneously expressing dominant-negative NSF. Treatment of E329Q-NSF-expressing cells with the ROCK inhibitor Y-27632 did inhibit FA formation. The results are consistent with a model of cell adhesion in which SNARE-mediated membrane traffic is required for both the elaboration of lamellipodia and the modulation of biochemical signals that control RhoA-mediated FA assembly. PMID- 16243315 TI - Inhibition of RhoA-mediated SRF activation by p116Rip. AB - p116Rip, originally identified as a binding partner of activated RhoA, is an actin-binding protein that interacts with the regulatory myosin-binding subunit (MBS) of myosin-II phosphatase and is essential for Rho-regulated cytoskeletal contractility. Here, we have examined the role of p116Rip in RhoA-mediated activation of the transcription factor SRF. We show that p116Rip oligomerizes via its C-terminal coiled-coil domain and, when overexpressed, inhibits RhoA-induced SRF activation without affecting RhoA-GTP levels. Mutant forms of p116Rip that fail to oligomerize or bind to MBS are still capable of inhibiting SRF activity. Our results suggest that p116Rip interferes with RhoA-mediated transcription through its ability to disassemble the actomyosin cytoskeleton downstream of RhoA. PMID- 16243316 TI - Cell integrity signaling activation in response to hyperosmotic shock in yeast. AB - Current progress highlights the role of the yeast cell wall as a highly dynamic structure that responds to many environmental stresses. Here, we show that hyperosmotic shock transiently activates the PKC signaling pathway, a response that requires previous activation of the HOG pathway. Phosphorylation of Slt2p under such conditions is related to changes in the glycerol turnover and is mostly Mid2p dependent, suggesting that changes in cell turgor, mediated by intracellular accumulation of glycerol, are sensed by PKC sensors to promote the cell integrity response. These observations, together with previous results, suggest that yeast cells respond to changes in cellular turgor by remodeling their cell walls. PMID- 16243318 TI - Functional zinc finger/sleeping beauty transposase chimeras exhibit attenuated overproduction inhibition. AB - The sleeping beauty (SB) transposon system has potential utility in gene transfer applications but lacks specificity for genomic integration and exhibits overproduction inhibition which limits in vivo activity. Targeting transposition may be possible by coupling a specific DNA binding domain to the SB transposase, but it is not known if this strategy will preserve or disrupt activity of the system. We engineered and tested chimeric SB transposases with two different human zinc finger DNA binding domain elements, Sp1 and zinc finger 202 (ZNF202). Addition of Sp1 to the C-terminus abolished transposase activity whereas N terminal addition of either Sp1 or ZNF202 did not. Transposition activity exhibited by N-terminal chimeras was increased to levels similar to native SB through the use of a hyperactive transposase (SB12) and activating transposon mutations. Importantly, addition of DNA binding domains to the transposase N terminus resulted in attenuation of overproduction inhibition, a major limitation of this system. These findings suggest that SB transposase chimeras may have specific advantages over the native enzyme. PMID- 16243317 TI - Low and high molecular weight poly(L-lysine)s/poly(L-lysine)-DNA complexes initiate mitochondrial-mediated apoptosis differently. AB - Poly(L-lysine)s, PLLs, are commonly used for DNA compaction and cell transfection. We report that, although PLLs of low (2.9 kDa), L-PLL, and high (27.4 kDa), H-PLL, Mw in free form and DNA-complexed cannot only cause rapid plasma membrane damage in human cell lines, phosphatidylserine "scrambling" and loss of membrane integrity, but later (24 h) initiate stress-induced cell death via mitochondrial permeabilization without the involvement of processed caspase 2. Mitochondrially mediated apoptosis was confirmed by detection of cytochrome c (Cyt c) release, activation of caspases-9 and -3, and subsequent changes in mitochondrial membrane potential. Plasma membrane damage and apoptosis were most prominent with H-PLL. Cytoplasmic level of Cyt c was more elevated following H PLL treatment, but unlike L-PLL case, inhibition of Bax channel-forming activity reduced the extent of Cyt c release from mitochondria by half. Inhibition of Bax channel-forming activity had no modulatory effect on L-PLL-mediated Cyt c release. Further, functional studies of isolated mitochondria indicate that H PLL, but not L-PLL, can directly induce Cyt c release, membrane depolarization, and a progressive decline in the rate of uncoupled respiration. Combined, our data suggest that H-PLL and L-PLL are capable of initiating mitochondrially mediated apoptosis differently. The observed PLL-mediated late-phase apoptosis may provide an explanation for previously reported transient gene expression associated with PLL-based transfection vectors. The importance of our data in relation to design of novel and safer cationic non-viral vectors for human gene therapy is discussed. PMID- 16243319 TI - Characterization of the dinuclear metal center of Pyrococcus furiosus prolidase by analysis of targeted mutants. AB - Prolidases are dipeptidases specific for cleavage of Xaa-Pro dipeptides. Pyrococcus furiosus prolidase is a homodimer having one Co-bound dinuclear metal cluster per monomer with one tightly bound Co(II) site and the other loosely bound (Kd 0.24 mM). To identify which Co site is tight-binding and which is loose binding, site-directed mutagenesis was used to modify amino acid residues that participate in binding the Co1 (E-313 and H-284), the Co2 site (D-209) or the bidentate ligand (E-327). Metal-content, enzyme activity and CD-spectra analyses of D209A-, H284L-, and E327L-prolidase mutants show that Co1 is the tight-binding and Co2 the loose-binding metal center. PMID- 16243320 TI - In situ and in vivo efficacy of peroral absorption enhancers in rats and correlation to in vitro mechanistic studies. AB - The present investigation attempts to increase intestinal permeability and hence absorption of biopharmaceutic classification system (BCS) Class III (cefotaxime sodium (CX)) and Class IV (cyclosporin A (CSA)) drugs by employing certain absorption enhancers. Drugs were co-perfused with sodium caprate (SC, 0.25% w/v), piperine (P, 0.004% w/v) and sodium deoxycholate (SD, 1.0% w/v) separately in rat in situ single pass intestinal perfusion model. These additives increased intestinal permeability (P(app)) and absorption rate constant (K(a)) up to two and fourfold, respectively. SC exhibited substantial absorption enhancement of both CX and CSA, while SD and P enhanced absorption of CX and CSA, respectively. Co-administration of SC significantly enhanced peroral bioavailability of CX (from 29.4 +/- 1.7 to 69.6 +/- 3.2) and CSA (from 18.4 +/- 15.6 to 49.6 +/- 25.1) in rats, while P increased bioavailability of CSA (from 18.4 +/- 15.6 to 33.1 +/- 17.7). Transmission electron microscopy of intestinal mucosa revealed that SC and SD act on lipid and protein domains of absorptive membrane. P showed no effect on intestinal P(app) and oral bioavailability of CX but has a profound effect on CSA, a known P-glycoprotein (P-gp) substrate. These results indicated that P enhances intestinal absorption of CSA by modulating P-gp mediated efflux transport. Release of lactate dehydrogenase in situ from intestinal mucosa in the presence of absorption enhancer was taken as index of its local toxicity. All the absorption enhancers showed significantly less release of LDH compared to positive control, sodium dodecyl sulfate (60% w/v). Overall, the data indicate that the features of these commonly used food ingredients or endogenous bile salts can effectively improve bioavailability of various BCS Class III and Class IV drugs. PMID- 16243321 TI - New 8-substituted xanthiene derivatives as potent bronchodilators. AB - The synthesis and structure determination of 8-aryl /alkyl aryl 1, 3-dimethyl-3, 7-dihydropurin-2, 6-dione derivatives (1-13), was carried out in this study. Bronchodilator activity is investigated using isolated guinea-pig tracheal strips, pre-contracted by acetylcholine and histamine. Spasmolytic activity of the compounds was compared to theophylline. Synthesized compounds (1-13) did not inhibit the acetylcholine-induced pre-contractions except compound (8) at 10(-5) M concentration. In contrast, some of the compounds, especially (7), (11), (12) at 10(-5) M and (3), (4), (9) and (11) in 10(-4) M displayed inhibitory activity on the tracheal strips pre-contracted by histamine. The potency of the compounds at human adenosine receptors was evaluated using radioligand binding assay and a cyclic AMP functional assay in CHO cells expressing these receptors. Compound (11) displayed the greatest activity against radioligand binding of specific agonists to A2A and A2B receptors. The compounds were relatively selective for both A2A and A2B compared with A1 and A3 receptors. All compounds were also tested for the inhibition of NECA-induced cAMP accumulation mediated by the A2B adenosine receptor and compound (11) was found to be the most effective. Our results showed that these compounds are acting as selective adenosine antagonists, especially for adenosine A2B receptors, and are promising as potent anti-inflammatory agents rather than bronchodilator for the treatment of asthma. PMID- 16243322 TI - Nasal administration of heparin-loaded microspheres based on poly(lactic acid). AB - In this study, heparin-loaded microspheres having smooth surface and small particle size were designed in order to provide the absorption of heparin through nasal mucosa. For this purpose, microspheres at different polymer/drug ratios (1:10, 1:2.5 and 1:1) and at different concentrations of polyvinyl alcohol, emulsifying agent (1.5% and 2.5% w/v) were prepared by solvent evaporation method with poly(lactic acid). The microspheres were for evaluated shape and surface properties, particle size, production yield, encapsulation efficiency and in vitro drug release. Based on the in vitro data, selected microspheres were applied by nasal route to Wistar albino rats. According to in vivo studies, heparin-loaded microspheres may be used by nasal route as an alternative to parenteral route. PMID- 16243323 TI - Chemical and biochemical transformations of 5-ethoxycarbonyl-5-phenyl-2 isoxazolines. AB - The salts, 2-methyl-5,5-disubstituted 4,5-dihydroisoxazolium methylsulfates comprising various substituents at the C-3 carbon atom were subjected to transformations. The structure of applied compounds permitted to monitor the effect of this factor on the transformation course of the 2-isoxazoline ring. The nucleophilic addition of cyanide anion to the selected salts enabled the obtaining of a next heterocyclic system of changed physicochemical and biological properties in comparison to the starting 2-isoxazolines. The diastereoselective hydrolysis of the cyanide group in 2-isoxazolidines by the bacteria strain Rhodococcus rhodochrous PCM 909 leads to the obtaining of a racemic mixture of the trans-hydroxyacid. The introduction of new functional groups into the heterocyclic ring made these compounds attractive objects for further chemical and microbial transformations and to study their biological activity. PMID- 16243325 TI - Integrative approach to osmoregulatory action of atrial natriuretic peptide in seawater eels. AB - Atrial natriuretic peptide (ANP) reduces plasma Na+ concentration and promotes seawater (SW) adaptation in SW eels. However, little is known about the mechanisms for the hyponatremic effect of ANP. In order to evaluate the role of ANP in the whole-body Na+ homeostasis of marine teleost, we reviewed previous in vivo experiments using exogenously administered ANP and present additional experiments to assess the role of endogenous ANP in Na+ homeostasis in conscious SW eels. The Na+ influx and efflux rate across the body surfaces including the gills measured with isotopic 22Na were not altered by the hyponatremic dose (5 pmol kg-1 min-1) of ANP infusion in SW eels. ANP infusion also had no effect on renal Na+ excretion in SW eels. In contrast, ANP strongly inhibited drinking, and the inhibition was quantitatively correlated with the hyponatremic effect of ANP. Further, intestinal absorption of Na+ was inhibited by ANP as examined in situ using intestinal sac in conscious SW eels. The combined inhibitory actions of ANP on drinking and intestinal absorption were sufficient to explain the decrease in plasma Na+ concentration. In addition, removal of endogenous circulating ANP by immunoneutralization increased plasma Na+ concentration with a concomitant increase in drinking rate in SW eels. These results strongly suggest that endogenous ANP is involved in the hyponatremic regulation through actions on drinking, and probably on intestine, in SW eels. PMID- 16243324 TI - Changes in mRNA expression of grouper (Epinephelus coioides) growth hormone and insulin-like growth factor I in response to nutritional status. AB - Growth hormone (GH) and insulin-like growth factor-I (IGF-I) are key links to nutritional condition and growth regulation in teleost. To understand the endocrine mechanism of growth regulation in grouper, we cloned the cDNAs for grouper GH and IGF-I and examined their mRNA expression during different nutritional status. Grouper GH cDNA is 936 base pairs (bp) long excluding the poly-A tail. It contained untranslated regions of 85 and 231bp in the 5'- and 3' ends, respectively. It has an open reading frame of 612bp coding for a signal peptide of 17 amino acids (aa) and a mature hormone of 187aa residues. Based on the aa sequence of the mature hormone, grouper GH shows higher sequence identity (>76%) to GHs of perciforms than to GHs of cyprinids and salmonids (53-69%). Grouper preproIGF-I cDNA consisted of 558bp, which codes for 186aa. This is composed of 44aa for the signal peptide, 68aa for the mature peptide comprising B, C, A, and D domains, and 74aa for the E domain. Mature grouper IGF-I shows very high sequence identity to IGF-I of teleost fishes (84-97%) compared to advanced groups of vertebrates such as chicken, pig, and human (80%). Using DNA primers specific for grouper GH and IGF-I, the changes in mRNA levels of pituitary GH and hepatic IGF-I in response to starvation and refeeding were examined by a semi-quantitative RT-PCR. Significant elevation of GH mRNA level was observed after 2 weeks of food deprivation, and increased further after 3 and 4 weeks of starvation. GH mRNA level in fed-controls did not change significantly during the same period. Hepatic IGF-I mRNA level decreased significantly starting after 1 week of starvation until the 4th week. There was no significant change in IGF-I mRNA levels in fed-controls. One week of refeeding can restore the GH and IGF-I mRNA back to its normal levels. Deprivation of food for 1-4 weeks also resulted in cessation of growth and decrease in condition factor. PMID- 16243326 TI - Melatonin and differential effect of L-thyroxine on immune system of Indian tropical bird Perdicula asiatica. AB - Interaction of thyroxine and melatonin on immune status was noted in vivo and in vitro when peripheral melatonin was high and thyroxine low in plasma of male Perdicula asiatica during reproductively inactive phase. During this phase exogenous thyroxine (4 microg/100g. Bwt./day) and melatonin (25 microg/100g. Bwt./day) increased immune parameters (spleen weight, total leukocyte count, lymphocyte count, percent stimulation ratio) and increased splenocyte density in spleen. In vitro l-thyroxine (10(-6)M/ml) supplementation decreased the splenocyte proliferation which was reversed by melatonin (500 pg/ml) supplementation. In vivo l-thyroxine showed immunoenhancing effect while in vitro it decreased the splenocyte proliferation presenting a differential effect. In the absence of internal physiological conditions of the birds, T(4) showed a negative effect on splenocytes proliferation in vitro when treated alone. However, melatonin maintained its lymphoproliferative effect under both conditions. Thus, avian splenocyte exposed to different hormonal conditions in vitro might have produced different signal peptides other than in vivo, thereby making the result different. PMID- 16243327 TI - Occurrence of two functionally distinct proopiomelanocortin genes in all modern lampreys. AB - The lampreys (family Petromyzontidae) are divided into three subfamilies, the Petromyzontinae in the Northern Hemisphere and the Geotriinae and Mordaciinae in the Southern Hemisphere. We previously found two proopiomelanocortin subtypes, proopiocortin (POC) and proopiomelanotropin (POM) in sea lamprey, Petromyzon marinus (Petromyzontinae). POC encoding adrenocorticotropic hormone (ACTH) and beta-endorphin (beta-END) is expressed in the pars distalis of the pituitary, while POM encoding melanophore-stimulating hormone (MSH)-A and B together with a different beta-END is expressed in the pars intermedia of the pituitary. All these hormonal segments are encoded on the third exon in both POC and POM. Here, we demonstrate the presence of both POC and POM genes in Geotria australis (Geotriinae) and Mordacia mordax (Mordaciinae) by molecular cloning of the third exons with the polymerase chain reaction using genomic DNA or pituitary cDNA. Molecular phylogenetic analysis showed that the POC and POM are distinctly different for the Southern Hemisphere lampreys as they are for P. marinus. Moreover, the relationship of each hormonal segments in POC and POM between Geotria, Mordacia, and Petromyzon is inconsistent. Immunocytochemical studies showed that the distribution of POC and POM in the pituitaries of the Southern Hemisphere lampreys is the same as that in the Northern Hemisphere. Taken together, these findings suggest that the duplication event which generated the two genes may have occurred in a common ancestor of the three extant lamprey subfamilies. PMID- 16243328 TI - Smoking and hip fracture; a study of 3617 cases. AB - We analysed the characteristics and outcome for 467 hip fracture patients, who reported that they were currently smoking at the time of admission, against 3150 non-smoking hip fracture patients. Those patients who smoked were younger (72 years versus 81 years mean age), more likely to be male (35.3% versus 19.5%), more mobile and less likely to be living in institutional care (7.5% versus 25.0%). Preoperative outcomes and complication rates were similar, despite the smokers' relative youth. Mortality at 30 days was similar for the two groups (6.2% versus 7.6%), but lower for the smokers at one year (22.7% versus 27.6%). However, with adjustment for the younger age and sex of these patients, this difference in mortality was not statistically significant. These findings suggest that smoking results in hip fracture occurring at a younger age. Despite this, the outcome for smokers was similar to that for the average hip fracture patient. PMID- 16243329 TI - Dynamic hip screw in the management of reverse obliquity intertrochanteric neck of femur fractures [Injury 36 (2005) 105-109]. PMID- 16243330 TI - "Superficial radial nerve damage due to Kirschner wiring of the radius" by Singh et al. [Injury 36 (2005) 330-332]. PMID- 16243331 TI - Traumatic lung injury treated by extracorporeal membrane oxygenation (ECMO). AB - BACKGROUND: Conventional mechanical ventilation is the mainstay of treatment for severe respiratory failure associated with trauma. However, when extensive lung injury is present, this technique may not be sufficient to prevent hypoxia, and furthermore, may exacerbate pulmonary damage by barotrauma. Extracorporeal membrane oxygenation (ECMO) has been used successfully in critically ill adult trauma patients and can offer an additional treatment modality. This study reports the use of ECMO in a cohort of adults referred with severe respiratory failure following trauma. METHODS: Retrospective analysis over an 8-year period of all 28 adult patients referred to a single tertiary unit for ECMO support. Survival relative to Injury severity score (ISS), lung injury score (Murray grade), duration of treatment and patient age was evaluated. RESULTS: Twenty of 28 patients who received ECMO with severe trauma related respiratory failure (mean PaO2/FiO2 of 62 mmHg) survived. Most patients had long bone fractures, blunt chest trauma, or combined injuries. Lung injury and injury severity scores, patient age, ECMO duration and oxygenation indices pre-ECMO (PaO2/FiO2) were similar in both the survivor and non-survivor groups. CONCLUSION: A high proportion of trauma patients treated with ECMO for severe lung injury survived. This outcome appears to compare favourably to conventional ventilation techniques and may have a role in patients who develop acute severe respiratory distress associated with trauma. PMID- 16243332 TI - Derotation of post-traumatic femoral deformities by closed intramedullary sawing. AB - Different techniques and devices have been used for correction osteotomies of bones in patients with malalignments. The most frequently used technique for rotational deformities of the femur and tibia is open osteotomy with an oscillating saw and pre-drilled holes with all well-known drawbacks of open surgery. An intramedullary device with an adapted minimal-invasive surgical technique allows intramedullary osteotomy of the bone preserving the surrounding soft tissue. We performed femoral osteotomies with an intramedullary saw followed by static interlocking nailing in 14 patients with post-traumatic rotational deformity in the femur. Twelve patients had an external rotational deformity of the femur ranging between 26 and 63 degrees , one had an additional leg shortening of about 4 cm. Two patients had internal rotational deformities. In two patients with delayed fracture healing union was achieved within one year without secondary surgery. Post-operative clinical assessment and CT-scans revealed good derotation results with deformities of less than 4 degrees in all cases. No device-related complications were observed. Therefore, we conclude that "closed" osteotomy with an intramedullary saw is a minimal-invasive, safe and reliable option for derotation procedures in the femur. PMID- 16243333 TI - Functional outcome of tibial plateau fractures treated with the fine-wire fixator. AB - OBJECTIVE: To review the functional outcome of patients with complex tibial plateau fractures treated with fine-wire fixation. DESIGN: Retrospective review with follow-up of patients in outpatient clinic. SETTING: Tertiary trauma center. PATIENTS: All patients who had fine-wire fixation for tibial plateau fractures between 1996 and January 2001 were reviewed. INTERVENTIONS: Fine-wire fixation with/without limited internal fixation for complex tibial plateau fractures. MAIN OUTCOME MEASURES: Knee range of motion, adequacy of articular surface reduction, mechanical axis, Knee Society Clinical Rating Scale and Short-Form 36 Health Questionnaire. RESULTS: Eighteen of twenty-one eligible patients were available for follow-up. There were 14 Shatzker VI and 4 V fractures. Seven fractures were open. Average follow-up was 28.2 months. All fractures united. There were three cases of delayed union, all progressed to union following additional procedures and bone grafting. There were no cases of osteomyelitis, septic arthritis or deep vein thrombosis. Seven patients had Knee Society Clinical Rating Scores of good/excellent (38.9%), and 11 had fair/poor scores (61.1%). Abnormal mechanical axes and multiple co-morbid injuries were associated with poorer outcomes. Although SF-36 scores were lower in the study group compared to matched population norms, 15 of 18 patients had full or partial return to pre-injury levels of functioning. CONCLUSION: Fine-wire fixation with limited internal fixation is a satisfactory method of managing complex high-energy fractures of the tibial plateau where soft tissue injury and bony comminution make traditional techniques of open reduction and internal fixation unsuitable. PMID- 16243334 TI - Treatment of infected defect pseudoarthrosis of the tibia by in situ fibular transfer in children. AB - The aim of this study was to determine the effectiveness of in situ fibular transfer in the treatment of infected defect pseudoarthrosis in childhood. Four cases with infected defect pseudoarthrosis of the tibia were treated with debridement, sequestrectomy and two-staged ipsilateral fibular transfer. The mean age of the patients was 7.5 (2-11) years. Mean follow-up period was 9.5 (6-13) years. In all cases infection was resolved and tibia-fibular synostosis was achieved. The diameter of the transferred fibula increased significantly with regard to the unaffected fibula. None of the patients had limitation of joint motion or shortening more than 1cm. This technique is a good method for treatment of infected defect pseudoarthrosis of the tibia. PMID- 16243336 TI - Guided bone regeneration in the treatment of segmental diaphyseal defects: a comparison between resorbable and non-resorbable membranes. AB - This study compares non-absorbable, ethyl cellulose (EC) membranes and absorbable, chitosan (CH) membranes in a rabbit model. Bone regeneration, evaluated by radiological and pathological parameters, was seen in both groups. In the EC group the process was faster but was followed by a sharp decrease, whereas in the CH group it was slower and continuous. PMID- 16243335 TI - Distraction osteogenesis for segmental bone defect. Physeal change after acute bone shortening followed by gradual lengthening in a rabbit tibia model. AB - We investigated physeal change after acute bone shortening followed by distraction osteogenesis in a rabbit tibia model. Distraction osteogenesis was performed on acutely shortened right rabbit tibia after removing a diaphyseal segment until distraction reached the length of the excised segment. Sixty animals were divided into three groups (n=20) according to the amount of shortening (20, 30, and 40% of original length) and radiographic, immunohistochemical, and histomorphometric analyses were performed to evaluate the physeal activity. The results showed no differences between groups and all animals could recover the original length without growth retardation. There was no notable venous congestion or lymphatic kinking associated with acute shortening. These results suggest that distraction osteogenesis at the primary site of shortening may have little adverse effects on the growth plate, when the shortening can provide sufficient soft tissue without vascular compromise and the amount of lengthening does not exceed that of shortening. PMID- 16243337 TI - The effectiveness of standard povidone iodine surgical preparation in decontaminating external fixator components. AB - The purpose of this study was to evaluate the effectiveness of standard iodine surgical scrubs to remove bacteria from external fixator components. Sterile adjustable external fixation clamps, Schanz pins, and carbon fibre rods were coated with a sterile protein solution and immersed in solution of coagulase negative Staphylococcus (10(3)organisms/ml). They were then decontaminated in standard fashion using a povidone iodine scrub and paint solution. After neutralisation the components were sonicated, serially diluted, plated on blood agar, and incubated for 24h. Unassembled external fixation components were examined individually, and as assembled pin-rod-clamp constructs with and without manipulation of the clamp. Of the three external fixation components (pins, rods, clamps) the highest number of bacterial colony forming units was seen on the external fixation clamps. Manipulation of the assembled construct significantly increased the mean bacterial colony counts compared to the assembled non manipulated construct (p=0.0007). Standard surgical preparation does not remove all bacteria from external fixators during subsequent operative procedures. PMID- 16243338 TI - Electrosurgical bipolar vessel sealing during vaginal hysterectomy. AB - OBJECTIVE: To compare electrosurgical bipolar vessel sealing (EBVS) with traditional suturing during vaginal hysterectomy. METHODS: In a randomized controlled trial involving 68 women undergoing vaginal hysterectomy for benign disease, 37 procedures were performed using EBVS and 31 using traditional suturing. The end points were procedure time, blood loss, number of ligatures used, postoperative pain score, and number of days in hospital. RESULTS: The procedure duration was shorter using EBVS (median duration, 32 vs. 40 min; P=.0003), with fewer ligatures (1 vs. 7; P<.0001) and less pain (median score, 4 vs. 6; P<.0001). There were no significant differences regarding blood loss (median, 100 vs. 160 mL; P=.36) and days in hospital (median, 2 vs. 2; P=.03). CONCLUSION: The EBVS system provided advantages over traditional suturing with regard to procedure time, number of ligatures used, and postoperative pain score. PMID- 16243339 TI - Calcium plus linoleic acid therapy for pregnancy-induced hypertension. AB - OBJECTIVE: To determine the effect of dietary supplementation of calcium plus conjugated linoleic acid (calcium-CLA) in reducing the incidence of vascular endothelial dysfunction in pregnant women at high risk of developing pregnancy induced hypertension (PIH). PATIENTS AND METHODS: This randomized, double-blind, placebo-controlled trial conducted at 4 outpatient clinics in 2 developing countries recruited 48 healthy primigravidas younger than 19 years or older than 35 years who had a family history of pre-eclampsia and diastolic notch. Twenty four participants received daily elemental calcium (600 mg) plus CLA (450 mg) and 24 received placebo from week 18 to 22 of pregnancy until delivery. RESULTS: Calcium-CLA supplementation reduced significantly the incidence of PIH (2 cases [8%] in the study group vs. 10 cases [42%] in the placebo group; relative risk, 0.20; 95% confidence interval, 0.05-0.82; P=.01). Endothelial dysfunction was also significantly reduced after calcium-CLA supplementation (in 18 women [75%] vs. 4 women [17%]; P<.001), compared with the placebo group (in 15 [63%] vs. 9 women [38%]; P=.08). CONCLUSION: In pregnant women at high risk for PIH, calcium CLA supplementation decreases the incidence of PIH and improves endothelial function. PMID- 16243340 TI - Association of TNF-alpha serum levels and TNFA promoter polymorphisms with risk of myocardial infarction. AB - Elevated levels of tumor necrosis factor-alpha (TNF-alpha), and presence of polymorphisms of the TNFA gene have been implicated in cardiovascular disease pathogenesis. We explored the relationship between polymorphisms in the TNFA gene (-1031C/T, -863C/A -857T/C, -308G/A, -238G/A), protein levels of TNF-alpha and their association to myocardial infarction (MI) using a sample of 1213 post-MI patients and 1561 healthy controls. MI risk was higher among men with elevated TNF-alpha levels, with the highest compared to the lowest TNF-alpha quartile giving a 70% risk increase (OR [95% CI]: 1.7 [1.1; 2.6]). Obese subjects who also had elevated TNF-alpha levels were at even higher risk for MI (OR [95% CI]: 3.4 [2.1; 5.6]). Higher TNF-alpha levels were seen among smokers (but not among non smokers) carrying the -857T allele. Furthermore, a rare haplotype occurred more frequently among the cases than the controls. Elevated TNF-alpha levels are associated with increased MI risk. Obese subjects with elevated TNF-a levels, and carriers of polymorphisms in or near TNFA are particularly susceptible to the hazards of smoking, results which may have implications for cardiovascular preventive measures. PMID- 16243342 TI - Determination of diallyldimethylammonium chloride in drinking water by reversed phase ion-pair chromatography-electrospray ionization mass spectrometry. AB - A method for the direct determination of diallyldimethylammonium chloride (DADMAC) in water samples, using ion-pair liquid chromatography-mass spectrometry system was developed. The chromatographic separation was performed using a C18 column. The type, the concentration of ion-pair reagent and the pH were optimized to give good chromatographic retention and sensitivity for DADMAC. Quantification was achieved in the positive electrospray ionization mode using selected ion monitoring. The cone voltage was also studied to establish the optimal experimental conditions. Finally, the reproducibility of the proposed method was shown by good run-to-run and day-to-day precision values. No sample preparation was required and the detection limit was 0.1 microg/L. The method was used to detect residual DADMAC at drinking water treatment plants in Tianjin, north China. The concentration of DADMAC observed in drinking water ranged from below quantitation limit to 22.0 microg/L. PMID- 16243341 TI - Influence of advanced electromyogram (EMG) amplitude processors on EMG-to-torque estimation during constant-posture, force-varying contractions. AB - Numerous studies have investigated the relationship between surface electromyogram (EMG) and torque exerted about a joint. Most studies have used conventional EMG amplitude (EMGamp) processing, such as rectification followed by low-pass filtering, to pre-process the EMG before relating it to torque. Recently, advanced EMGamp processors that incorporate signal whitening and multiple-channel combination have been shown to significantly improve EMGamp processing. In this study, we compared the performance of EMGamp-torque estimators with and without these advanced EMGamp processors. Fifteen subjects produced constant-posture, non-fatiguing, force-varying contractions about the elbow while torque and biceps/triceps EMG were recorded. EMGamp was related to torque using a linear FIR model. Both whitening and multiple-channel combination reduced EMG-torque errors and their combination provided an additive benefit. Using a 15th-order linear FIR model, EMG-torque errors with a four-channel, whitened processor averaged 7.3% of maximum voluntary contraction (MVC) (or 78% of variance accounted for). By comparison, the equivalent single-channel, unwhitened (conventional) processor produced an average error of 9.9% of MVC (variance accounted for of 55%). In addition, the study describes the occurrence of spurious peaks in estimated torque when the torque model is created from data with a sampling rate well above the bandwidth of the torque. This problem occurs when the torque data are sampled at the same rate as the EMG data. The problem is corrected by decimating the EMGamp prior to relating it to joint torque, in our case to an effective sampling rate of 40.96 Hz. PMID- 16243343 TI - Separation of conjugated linoleic acid isomers by cyclodextrin-modified micellar electrokinetic chromatography. AB - A cyclodextrin-modified micellar electrokinetic chromatography (CD-MEKC) method was developed for separating conjugated linoleic acid (CLA) isomers. All the seven CLA isomers (9cis,11cis-CLA, 9cis,11trans-CLA, 9trans,11trans-CLA, 10trans,12cis-CLA, 11cis,13cis-CLA, 11cis,13trans-CLA and 11trans,13trans-CLA) were completely separated in the optimized conditions (4% (w/v) beta-cyclodextrin (beta-CD), 54 mM sodium dodecyl sulphate (SDS), 80 mM borate (pH 9.0), 8 M urea, 4% (v/v) ethanol, 30 kV and 15 degrees C). The CD-MEKC method was superior to the gas chromatographic (GC) and silver-ion high-performance liquid chromatographic (Ag(+)-HPLC) methods that were generally used in analyzing CLA isomers. PMID- 16243345 TI - Interaction forces and molecular adhesion between pre-adsorbed poly(ethylene imine) layers. AB - Interaction forces between pre-adsorbed layers of branched poly(ethylene imine) (PEI) of different molecular mass were studied with the colloidal probe technique, which is based on atomic force microscopy (AFM). During approach, the long-ranged forces between the surfaces are repulsive due to overlap of diffuse layers down to distances of a few nanometers, whereby regulation of the surface charge is observed. The ionic strength dependence of the observed diffuse layer potentials can be rationalized with a surface charge of 2.3 mC/m2. The forces remain repulsive down to contact, likely due to electro-steric interactions between the PEI layers. These electro-steric forces have a range of a few nanometers and appear to be superposed to the force originating from the overlap of diffuse layers. During retraction of the surfaces, erratic attractive forces are observed due to molecular adhesion events (i.e., bridging adhesion). The frequency of the molecular adhesion events increases with increasing the ionic strength. The force response of the PEI segments is dominated by rubber-like extension profiles. Strong adhesion forces are observed for low molecular mass PEI at short distances directly after separation, while for high molecular mass weaker adhesion forces at larger distances are more common. The work of adhesion was estimated by integrating the retraction force profiles, and it was found to increase with the ionic strength. PMID- 16243344 TI - In situ neutron reflectometry investigation of gold-chemisorbed PEO layers of varying chain density: relationship of layer structure to protein resistance. AB - In work reported previously [L.D. Unsworth, H. Sheardown, J.L. Brash, Langmuir 21 (2005) 1036] we investigated protein interactions with polyethylene oxide (PEO) layers formed by chemisorption of thiol-PEO on gold. It was shown that, as a function of surface chain density, protein adsorption passed through a minimum. In follow-on work reported here, neutron reflectometry (NR) was used to investigate the formation and properties (volume fraction and chain density) of such PEO layers in situ, i.e., in contact with water. Chain density was varied by varying solubility conditions (far from and near the cloud point) and chemisorption time. Neutron experiments were carried out using neutrons of de Broglie wavelength 2.37 A. Contrast matching techniques were used to improve sensitivity. Layers formed under high solubility conditions were found to have PEO volume fraction, layer thickness and chain density of 0.33, 28 A, and 0.56 chains/nm2, respectively, after 0.5 h chemisorption; and 0.31, 28.5 A, and 0.59 chains/nm2, respectively, after 11 h, suggesting that the layer is fully formed within 0.5 h. Both chain density and PEO volume fraction in the chemisorbed layers were significantly greater when the layers were formed under low solubility conditions. The PEO layers shown in our previous work to have maximum protein resistance were found to have a PEO volume fraction of approximately 40%. Moreover the limiting volume fraction in the PEO films formed under low solubility conditions was approximately 57%, a value similar to the solubility limit of PEO in aqueous solution, suggesting that local regions in the layers may be phase separated under these conditions. This may result in increased hydrophobicity and may explain why protein adsorption was found to increase on the layers of higher chain density. PMID- 16243346 TI - Redispersible rutile TiO2 nanocrystals in organic media by surface chemical modification with an inorganic barium hydroxide. AB - The present paper describes the synthesis of the redispersible rutile TiO2 nanocrystals in organic media by surface chemical modification reaction in an aqueous barium hydroxide solution. In our facile surface modification reactions, the surfaces of the TiO2 nanocrystals are coated by bimetallic TiOBa spices and saturated with BaOH terminal groups. The inherent characteristics such as morphology, size, crystallinity, and color of the nanocrystals remained almost unchanged after surface-treatment, but their dispersibility in organic media such as methanol and DMF were remarkably enhanced. It is ascribed that BaOH groups in the surface of the TiO2 nanocrystals prevented the formation of covalently bound agglomerates through Ti-O-Ti condensation reaction among the nanocrystals during the purification and water-elimination procedures. PMID- 16243347 TI - Variations of wettability and protein adsorption on solid siliceous carriers grafted with poly(N-isopropylacrylamide). AB - Poly(N-isopropylacrylamide), a thermally responsive polymer, was end-grafted to mercaptopropyl derivatives of silica gel, plane glass sheets and glass capillary tubing by free radical polymerization of the monomer in 1,4-dioxane at 100 degrees C. The polymer monolayer attached to the glass carriers provided them with thermally controlled wettability registered by two independent methods: direct measurements of the water contact angle and capillary rise. The water contact angle changed from 54+/-3 degrees to 68+/-3 degrees in the temperature range from 20 to 50 degrees C. The polymer grafting to silica gel (pore diameter 100 A, particle size 5 microm) resulted in 15-30-fold reduction in protein adsorption on the carrier at 35 degrees C. Adsorption isotherms of myoglobin indicate completely different characters of the protein adsorption to silica gel and its polyNIPAM-grafted derivative. Cooling of the grafted carrier containing adsorbed myoglobin to 9 degrees C led to a partial release of the protein to the contacting solution, whereas heating of the system to 35 degrees C resulted in reversible binding of the protein. Adsorption of myoglobin on polyNIPAM-coated silica was ca. 2-fold higher at 35 than at 9 degrees C, most probably due to steric repulsion displayed by the swollen copolymer at the lower temperature. PMID- 16243348 TI - Stability and change in children's division strategies. AB - Age-related changes in children's performance on simple division problems (e.g., 6/2, 72/9) were investigated by asking children in Grades 4 through 7 to solve 32 simple division problems. Differences in performance were found across grade, with younger children performing more slowly and less accurately than older children. Problem size effects were also found in that children were faster and more accurate on small problems than on large problems. Two strategies changed across age, with children in Grade 4 relying heavily on the strategy of "addition" (adding the divisor until the dividend was reached) to solve the problems and children in Grades 5 through 7 relying primarily on the strategy of "multiplication" (recasting the division problem as a multiplication problem) to solve the problems. Surprisingly, the frequency of direct retrieval (retrieving the answer directly from memory) did not increase across grade and never became the dominant strategy of choice. Reasons for why retrieval use remains infrequent and age invariant are discussed. Overall, the results suggest that division is a unique operation and that the continued study of division may have implications for further understanding of how procedural and conceptual knowledge of arithmetic develops. PMID- 16243349 TI - Recognition of faces of ingroup and outgroup children and adults. AB - People are often more accurate in recognizing faces of ingroup members than in recognizing faces of outgroup members. Although own-group biases in face recognition are well established among adults, less attention has been given to such biases among children. This is surprising considering how often children give testimony in criminal and civil cases. In the current two studies, Euro Canadian children attending public school and young adults enrolled in university level classes were asked whether previously presented photographs of Euro American and African American adults (Study 1) or photographs of Native Canadian, Euro-Canadian, and African American children (Study 2) were new or old. In both studies, own-group biases were found on measures of discrimination accuracy and response bias as well as on estimates of reaction time, confidence, and confidence-accuracy relations. Results of both studies were consistent with predictions derived from multidimensional face space theory of face recognition. Implications of the current studies for the validity of children's eyewitness testimony are also discussed. PMID- 16243350 TI - Degradation of the S. frugiperda peritrophic matrix by an inducible maize cysteine protease. AB - A unique 33-kDa cysteine protease (Mir1-CP) rapidly accumulates at the feeding site in the whorls of maize (Zea mays L.) lines that are resistant to herbivory by Spodoptera frugiperda and other lepidopteran species. When larvae were reared on resistant plants, larval growth was reduced due to impaired nutrient utilization. Scanning electron microscopy (SEM) indicated that the peritrophic matrix (PM) was damaged when larvae fed on resistant plants or transgenic maize callus expressing Mir1-CP. To directly determine the effects of Mir1-CP on the PM in vitro, dissected PMs were treated with purified, recombinant Mir1-CP and the movement of Blue Dextran 2000 across the PM was measured. Mir1-CP completely permeabilized the PM and the time required to reach full permeability was inversely proportional to the concentration of Mir1-CP. Inclusion of E64, a specific cysteine protease inhibitor prevented the damage. The lumen side of the PM was more vulnerable to Mir1-CP attack than the epithelial side. Mir1-CP damaged the PM at pH values as high as 8.5 and more actively permeabilized the PM than equivalent concentrations of the cysteine proteases papain, bromelain and ficin. The effect of Mir1-CP on the PMs of Helicoverpa zea, Danaus plexippus, Ostrinia nubilalis, Periplaneta americana and Tenebrio molitor also was tested, but the greatest effect was on the S. frugiperda PM. These results demonstrate that the insect-inducible Mir1-CP directly damages the PM in vitro and is critical to insect defense in maize. PMID- 16243351 TI - Trimetazidine inhibits mitochondrial permeability transition pore opening and prevents lethal ischemia-reperfusion injury. AB - Trimetazidine (TMZ) affects mitochondrial function during ischemia. Mitochondrial permeability transition is a pivotal event in cardiomyocyte death following acute ischemia. The aim of the present study was to determine whether the anti-ischemic agent TMZ might modulate mitochondrial permeability transition pore (mPTP) opening and limit lethal ischemia-reperfusion injury. Anesthetized NZW rabbits underwent 30 min of coronary artery occlusion followed by 4 hours of reperfusion. Prior to this, they underwent either no intervention (control, C), ischemic preconditioning (PC), or an IV injection of 5 mg kg(-1) TMZ 10 min before ischemia (TMZ). Additional rabbits (Sham group) underwent no ischemia/reperfusion throughout the experiment. Infarct size was assessed by triphenyltetrazolium staining, and apoptosis via measurement of caspase 3 activity. Ca(2+)-induced mPTP opening was assessed in mitochondria isolated from ischemic myocardium. TMZ and PC significantly reduced infarct size that averaged 34 +/- 4% and 21 +/- 4% of the risk region respectively, versus 63 +/- 6% in controls (P<0.005). Caspase 3 activity was reduced in both TMZ and PC groups: 37 +/- 11 and 29 +/- 7 respectively, versus 68 +/- 9 nmol min(-1) mg(-1) mitochondrial protein in controls (P=0.01 versus TMZ and PC). In controls, Ca(2+) load required for mPTP opening averaged 11 +/- 4 microM mg(-1) mitochondrial protein versus 116 +/- 6 in shams (P<0.0001). Pre-treatment by TMZ or PC attenuated this, with Ca(2+) loads averaging 45 +/- 4 and 46 +/- 4 microM mg(-1) mitochondrial proteins, respectively (P<0.005 versus C). These data suggest that TMZ inhibits mPTP opening and protects the rabbit heart from prolonged ischemia-reperfusion injury. PMID- 16243352 TI - Helical packing of needles from functionally altered Shigella type III secretion systems. AB - Gram-negative bacteria commonly interact with eukaryotic host cells using type III secretion systems (TTSSs or secretons), which comprise cytoplasmic, transmembrane and extracellular domains. The extracellular domain is a hollow needle-like structure protruding 60 nm beyond the bacterial surface. The TTSS is activated to transfer bacterial proteins directly into a host cell only upon physical contact with the target cell. We showed previously that the monomer of the Shigella flexneri needle, MxiH, assembles into a helical structure with parameters similar to those defining the architecture of the extracellular components of bacterial flagella. By analogy with flagella, which are known to exist in different helical states, we proposed that changes in the helical packing of the needle might be used to sense host cell contact. Here, we show that, on the contrary, mutations within MxiH that lock the TTSS into altered secretion states do not detectably alter the helical packing of needles. This implies that either: (1) host cell contact is signalled through the TTSS via helical changes in the needle that are significantly smaller than those linked to structural changes in the flagellar filament and therefore too small to be detected by our analysis methods or (2) that signal transduction in this system occurs via a novel molecular mechanism. PMID- 16243353 TI - Measurement of internal movements within the 30 S ribosomal subunit using Forster resonance energy transfer. AB - We have used Forster resonance energy transfer (FRET) to study specific conformational changes in the Escherichia coli 30 S ribosomal subunit that occur upon association with the 50 S subunit. By measuring energy transfer between 13 different pairs of fluorescent probes attached to specific positions on 30 S subunit proteins, we have monitored changes in distance between different locations within the 30 S subunit in its free and 50 S-bound states. The measured distance changes provide restraints for modeling the movement that occurs within the 30 S subunit upon formation of the 70 S ribosome in solution. Treating the head, body, and platform domains of the 30 S subunit as simple rigid bodies, the lowest-energy solution converges on a model that satisfies each of the individual FRET restraints. In this model, the 30 S subunit head tilts towards the 50 S subunit, similar to the movement found in comparing 30 S subunits and 70 S ribosomes from X-ray and cryo-electron microscope structures, and the platform is predicted to undergo a clock-wise rotation upon association. PMID- 16243354 TI - ClpXP-dependent proteolysis of FNR upon loss of its O2-sensing [4Fe-4S] cluster. AB - The global regulator FNR from Escherichia coli controls the transcription of genes required for an anaerobic lifestyle. While previous studies have demonstrated that FNR activity is regulated by O2 through loss of dimerization upon destruction of its [4Fe-4S]2+ cluster, the present study reveals that monomeric FNR protein is also a target of proteolysis. We have found that turnover of FNR protein is increased selectively under aerobic growth conditions, when FNR is not active as a transcription factor and is primarily a metal-free, monomeric form (apo-FNR). This degradation of monomeric FNR was dependent on the ClpXP protease and required the presence of two amino acid sequences within FNR that resemble known ClpX recognition motifs. By measuring the turnover rates of various FNR mutants that have unique properties with respect to dimerization and Fe-S cluster stability, we have shown that loss of dimerization upon [4Fe-4S]2+ cluster destruction by O2 targets FNR for degradation by the ClpXP protease. In addition, by measuring the differential rate of FNR degradation upon switching aerobic cultures to anaerobic growth conditions, we provide evidence that pre existing FNR apo-protein can be converted to [4Fe-4S]2+ -FNR. Finally, we address the physiological significance of FNR proteolysis by demonstrating that varying FNR protein levels over a small range under aerobic growth conditions has a direct effect on the function of FNR in O2 sensing. PMID- 16243355 TI - A binding free energy hot spot in the ankyrin repeat protein GABPbeta mediated protein-protein interaction. AB - The frequently observed ankyrin repeat motif represents a structural scaffold evolved for mediating protein-protein interactions. As such, these repeats modulate a diverse range of cellular functions. We thermodynamically characterized the heterodimeric GA-binding protein (GABP) alphabeta complex and focused specifically on the interaction mediated by the ankyrin repeat domain of the GABPbeta. Our isothermal titration calorimetric analysis of the interaction between the GABP subunits determined an association constant (K(A)) of 6.0 x 10(8) M(-1) and that the association is favorably driven by a significant change in enthalpy (DeltaH) and a minor change in entropy (-TDeltaS). A total of 16 GABPbeta interface residues were chosen for alanine scanning mutagenesis. The calorimetrically measured differences in the free energy of binding were compared to computationally calculated values resulting in a correlation coefficient r = 0.71. We identified three spatially contiguous hydrophobic and aromatic residues that form a binding free energy hot spot (DeltaDeltaG > 2.0 kcal/mol). One residue provides structural support to the hot spot residues. Three non-hot spot residues are intermediate contributors (DeltaDeltaG approximately 1.0 kcal/mol) and create a canopy-like structure over the hot spot residues to possibly occlude solvent and orientate the subunits. The remaining interface residues are located peripherally and have weak contributions. Finally, our mutational analysis revealed a significant entropy-enthalpy compensation for this interaction. PMID- 16243356 TI - Trim32 is a ubiquitin ligase mutated in limb girdle muscular dystrophy type 2H that binds to skeletal muscle myosin and ubiquitinates actin. AB - Trim32 belongs to the tripartite motif (TRIM) protein family, which is characterized by a common domain structure composed of a RING-finger, a B-box, and a coiled-coil motif. In addition to these motifs, Trim32 possesses six C terminal NHL-domains. A point mutation in one NHL domain (D487N) has been linked to two forms of muscular dystrophy called limb girdle muscular dystrophy type 2H and sarcotubular myopathy. In the present study we demonstrate that Trim32 is an E3 ubiquitin ligase that acts in conjunction with ubiquitin-conjugating enzymes UbcH5a, UbcH5c, and UbcH6. Western blot analysis showed that Trim32 is expressed primarily in skeletal muscle, and revealed its differential expression from one muscle to another. The level of Trim32 expression was elevated significantly in muscle undergoing remodeling due to changes in weight bearing. Furthermore, expression of Trim32 was induced in myogenic differentiation. Thus, variability in Trim32 expression in different skeletal muscles could be due to induction of Trim32 expression upon changes in physiological conditions. We show that Trim32 associates with skeletal muscle thick filaments, interacting directly with the head and neck region of myosin. Our data indicate that myosin is not a substrate of Trim32; however, Trim32 was found to ubiquitinate actin in vitro and to cause a decrease in the level of endogenous actin when transfected into HEK293 cells. In conclusion, our results demonstrate that Trim32 is a ubiquitin ligase that is expressed in skeletal muscle, can be induced upon muscle unloading and reloading, associates with myofibrils and is able to ubiquitinate actin, suggesting its likely participation in myofibrillar protein turnover, especially during muscle adaptation. PMID- 16243358 TI - Coexistence and error propagation in pre-biotic vesicle models: a group selection approach. AB - Compartmentalization of unlinked, competing templates is widely accepted as a necessary step towards the evolution of complex organisms. However, preservation of information by templates confined to isolated vesicles of finite size faces much harder obstacles than by free templates: random drift allied to mutation pressure wipe out any template that does not replicate perfectly, no matter how small the error probability might be. In addition, drift alone hinders the coexistence of distinct templates in a same compartment. Here, we investigate the conditions for group selection to prevail over drift and mutation and hence to guarantee the maintenance and coexistence of distinct templates in a vesicle. Group selection is implemented through a vesicle survival probability that depends on the template composition. By considering the limit case of an infinite number of vesicles, each one carrying a finite number of templates, we were able to derive a set of recursion equations for the frequencies of vesicles with different template compositions. Numerical iteration of these recursions allows the exact characterization of the steady state of the vesicle population-a quasispecies of vesicles-thus revealing the values of the mutation and group selection intensities for which template coexistence is possible. Within the main assumption of the model-a fixed, finite or infinite, number of vesicles-we find no fundamental impediment to the coexistence of an arbitrary number of template types with the same replication rate inside a vesicle, except of course for the vesicle capacity. Group selection in the form of vesicle selection is a must for compartmentalized primordial genetic systems even in the absence of intra-genomic competition of different templates. PMID- 16243357 TI - Presenting characteristics of depressed outpatients as a function of recurrence: preliminary findings from the STAR*D clinical trial. AB - OBJECTIVES: Recurrent depression predicts risk for subsequent episodes, but it is unclear how it relates to demographic features, course of illness, and clinical presentation. METHODS: We report on the baseline data for the first 1500 patients enrolled in the Sequenced Treatment Alternatives to Relieve Depression (STAR*D) study (www.star-d.org). Patients were required to have a DSM-IV diagnosis of nonpsychotic major depression and to score > or = 14 on the 17-item Hamilton rating scale for depression. Status with respect to recurrent depression and other aspects of illness course and demographic features were ascertained at intake, along with measures of depression and concurrent general medical illness. RESULTS: Patients with recurrent depression were older, had an earlier age of onset, and were more likely to have a positive family history of depression than first episode patients. However, recurrent patients were less likely to be chronic and reported shorter current episodes than first episode patients, something that was largely confined to females. Recurrent patients were more likely than first episode patients to report non-essential aspects of mood, cognition, and somatic symptoms, although largely as a consequence of greater overall depressive symptom severity. CONCLUSIONS: As compared to single episode depressions, recurrent depression was associated with greater symptom severity and illness characteristics suggestive of greater underlying risk, but not other demographic characteristics than age. Risk for recurrence appeared to be distinct from chronic depression. A subset of chronic first episode patients may lack the capacity to remit and may therefore be distinct from those with recurrent episodes. PMID- 16243359 TI - Distributive network model of Banavar, Damuth, Maritan and Rinaldo (2002): critique and perspective. PMID- 16243360 TI - New perspectives on natural products in TB drug research. AB - The challenge of discovering new, urgently needed anti-TB drugs from natural sources requires a truly interdisciplinary research. Cutting-edge mycobacteriology and innovative natural products chemistry tools have to be developed and employed in tandem, in order to meet these demands. The present review provides cross-linkage to the most recent literature on anti-TB active natural products and summarizes the recent developments in both fields and their potential to impact the early steps of the TB drug discovery process. PMID- 16243361 TI - A comprehensive study on neurobehavior, neurotransmitters and lymphocyte subsets alteration of Chinese manganese welding workers. AB - The neurotoxicity of manganese has been demonstrated by many researches. But few reports have been found on its immunotoxicity in manganese-exposed workers. Here we selected welding workers (aged 34 years) as Mn-exposed subjects. They have been exposed to manganese for 16 years. The control group was from a flour plant. The average concentrations of Mn, Cd, Fe and Ni in work place were 138.40 +/- 11.60 microg/m3, 581.40 +/- 45.32 microg/m3, 3.84 +/- 0.53 microg/m3 and 12.64 +/ 2.80 ng/m3, respectively. Blood Mn (4.84 mug/dl) of welding workers was higher than that of the control group (1.92 microg/dl). Neurobehavioral core test battery (NCTB) recommended by WHO was conducted on the subjects and found that the scores of negative emotions, such as confusion-bewilderment, depression dejection, fatigue-inertia, and tension-anxiety, were higher in welding workers. Visual simple reaction time and the fast simple reaction time were shorter than that of the control group. The numbers of digital span, forward digital span, backward digital span and digital symbol decreased in welding workers compared with control group. Monoamine neurotransmitters and their metabolism substances in urine were tested by HPLC-ultraviolet. NE, E, MHPG, HVA, DA, DOPAC and 5-HT in the urine of Mn-exposed group had no significant changes while 5-HIAA in Mn exposed group had significantly decreased compared with that of the control group. Lymphocyte subsets of the subjects were determined by Flow Cytometer. CD3+ T cell, CD4+CD8- T cell, CD4-CD8+ T cell, CD4+CD45RO- "virgin" lymphocytes, CD4+CD45RO+ "memory" lymphocytes, and CD3-CD19+ B cell had no significant changes compared with the control group. The results showed that long-term exposure to manganese in welding might have adverse effects on mood state, neurobehavior, and peripheral neurotransmitters. However, they had no effects on lymphocyte subsets parameters. PMID- 16243362 TI - Nimesulide inhibits lipopolysaccharide-induced production of superoxide anions and nitric oxide and iNOS expression in alveolar macrophages. AB - The study was designed to investigate the effect of nimesulide on lipopolysaccharide (LPS)-induced proinflammatory oxidants production by rat alveolar macrophages (AMs). Effects of LPS and nimesulide on antioxidant defense and the expression of inducible nitric oxide synthase (iNOS) were also studied. It was found that nimesulide could scavenge superoxide anions (O2*-), nitric oxide (NO*) and total oxidant burden induced by LPS in AMs in vitro. Approximately 850 nmoles of nimesulide had activity equivalent to one IU of superoxide dismutase (SOD). Further, to confirm the in vitro observation, Male Wistar rats were orally administered with nimesulide (9 mg/kg b. wt. twice daily) for one week followed by intratracheal instillation of 2 microg LPS to stimulate lung inflammation. AMs from bronchoalveolar lavage fluid were collected 18 h after instillation of LPS. Nimesulide pretreatment could inhibit O2*-, NO() and lipid peroxidation in AMs. Nimesulide also suppressed LPS-induced iNOS expression in AMs in vivo and in vitro. Nimesulide could also normalize LPS-induced changes in the levels of superoxide dismutase (SOD), glutathione reductase (GR) and reduced glutathione (GSH) in AMs. Inhibition in production of oxidants in LPS challenged AMs by nimesulide could be one of the pathways for its anti inflammatory action. PMID- 16243363 TI - Occurrence of estrogen-like substances in the marine environment of the Northern Mediterranean Sea. AB - Evidence has been presented that a number of environmental agents perturbs the function of the sex hormone signalling pathways in marine animals. In this work the estrogenic/antiestrogenic activity of coastal marine water samples from five sites of the Northern Mediterranean Sea was assayed using a recombinant Saccharomyces cerevisiae yeast strain (RMY326 ER-ERE) transfected with the human estrogen receptor alpha. Extraction of potential EDCs from seawater was performed in columns packed with XAD-2 resins. The estrogenic activity was calculated as percentage of the activity of 17beta-estradiol (10 nM) and it ranged from 4.8% and 59.03%, significantly depending on sampling site (p=0.0013) and season (p<0.05). Antagonistic activity of extracts was also detected and the percentage of inhibition of estradiol-dependent beta-galactosidase induction ranged to 52.8%. These results point the reliability of the yeast assay as a first level screening test to assess the quality of aquatic environments. PMID- 16243364 TI - Galanin receptor-1 knockout mice exhibit spontaneous epilepsy, abnormal EEGs and altered inhibition in the hippocampus. AB - Galanin is a widely-distributed neuropeptide that acts as an endogenous anticonvulsant. We have recently generated a galanin receptor type 1 knockout mouse (Galr1(-/-)) that develops spontaneous seizures. Our aim here was to characterize the seizures by making electroencephalogram (EEG) recordings from this animal, and also to elucidate the cellular basis of its epileptic phenotype by studying the neurophysiology of CA1 pyramidal neurons in acute hippocampal slices. EEGs showed that major seizures had a partial onset with secondary generalization, and that paroxysms of spike-and-slow waves occurred and were associated with hypoactivity. The interictal EEG was also abnormal, with a marked excess of spike-and-slow waves. Slice experiments showed that resting potential, input resistance, intrinsic excitability, paired-pulse facilitation of excitatory and inhibitory postsynaptic currents (EPSCs and IPSCs), stimulus--response plots for EPSCs, and several properties of spontaneous miniature EPSCs and IPSCs were all unchanged in the mutant mouse compared with wildtype. However, the frequency of miniature IPSCs was significantly reduced in the mutants. These results suggest that impaired synaptic inhibition in the hippocampus may contribute to the local onset of seizures in the Galr1(-/-) mouse. PMID- 16243365 TI - On the nature of near space: effects of tool use and the transition to far space. AB - Many researchers have proposed that the near space immediately surrounding the body is represented differently than more distant space. Indeed, it has often been suggested that near space encompasses that within arm's reach. The present study used a line bisection task in healthy adults to investigate the effects of tool use on space perception, and the nature of the transition between near and far space. Subjects bisected lines at four distances controlled for both veridical and angular size using a laser pointer and a set of sticks. When the laser pointer was used, a left to right shift in bias was observed as stimuli were moved from near to far space. When a tool was used, however, a leftward bias was observed at all distances, similar to that observed with the laser pointer in near space. These results suggest that the tool expanded the range of near space. Additionally, the transition from near to far space was gradual, with no abrupt shift at arm's length (or at any other distance). In contrast to theories describing near space as that within arm's reach, these findings suggest that the representation of near space is less rigid, extending with tool use and gradually transitioning into far space. PMID- 16243366 TI - Normative influences on food intake. AB - Hunger and satiety have conventionally provided the framework for understanding eating and overeating. We argue that hunger and satiety play a relatively small role in everyday eating. The normative control of food intake refers to the fact that our eating is largely governed by the motive to avoid eating excessively. Dieters impose a restrictive intake norm on themselves, but often violate the norm. Personal norms are individualized rules that people develop to help themselves decide how much is appropriate to eat in a given situation. Situational norms are derived from the eating situation itself; examples include portion size and social influence, which exert powerful effects on intake. We discuss the implications of a normative approach to the analysis of eating and overeating. PMID- 16243367 TI - Effects of age on recovery of body weight following REM sleep deprivation of rats. AB - Chronically enforced rapid eye (paradoxical) movement sleep deprivation (REM-SD) of rats leads to a host of pathologies, of which hyperphagia and loss of body weight are among the most readily observed. In recent years, the etiology of many REM-SD-associated pathologies have been elucidated, but one unexplored area is whether age affects outcomes. In this study, male Sprague-Dawley rats at 2, 6, and 12 months of age were REM sleep-deprived with the platform (flowerpot) method for 10-12 days. Two-month-old rats resided on 7-cm platforms, while 10-cm platforms were used for 6- and 12-month-old rats; rats on 15-cm platforms served as tank controls (TCs). Daily changes in food consumption (g/kg(0.67)) and body weight (g) during baseline, REM-SD or TCs, and post-experiment recovery in home cages were determined. Compared to TCs, REM-SD resulted in higher food intake and decreases in body weight. When returned to home cages, food intake rapidly declined to baseline levels. Of primary interest was that rates of body weight gain during recovery differed between the age groups. Two-month-old rats rapidly restored body weight to pre-REM-SD mass within 5 days; 6-month-old rats were extrapolated by linear regression to have taken about 10 days, and for 12-month old rats, the estimate was about 35 days. The observation that restoration of body weight following its loss during REM-SD may be age-dependent is in general agreement with the literature on aging effects on how mammals respond to stress. PMID- 16243368 TI - Influence of early dietary experience on energy regulation in rats. AB - The environmental mechanisms that have contributed to the rapid increases in overweight and obesity in the US over the past 25-30 years have yet to be fully specified. One hypothesis that has been forwarded is that increased consumption of calories in liquid form may be a contributing factor, since some studies support the idea that caloric compensation is less adequate for liquid calories compared to calories delivered in more solid form. Work from our laboratory using rats has examined the role that differences in diet viscosity may play in altering energy intake and body weight regulation. This work has suggested that when offered diets matched on caloric density, macronutrient and micronutrient composition, and differing only in viscosity, adult rats fail to compensate for calories delivered in low-viscosity form in short-term intake tests. Further, long-term consumption of low-viscosity diets leads to enhanced weight gain in adult rats. In the present studies, we examined whether short- or long-term exposure to varying relationships between viscosity and calories led to altered food intake or body weight regulation in juvenile rats. The results demonstrated that animals given either short- or long-term experience with direct relationships between viscosity and calories (high viscosity, high calorie meals and low viscosity, low calorie meals) did not differ in food intake or body weight gain compared to animals given short- or long-term experience with indirect relationships between viscosity and calories (high viscosity, low calorie meals and low viscosity, high calorie meals). When juvenile rats were given long-term (9 weeks) exposure to a single, high or low viscosity diet supplement, matched on caloric density and differing only in viscosity, there were no effects on body weight gain. However, analysis of body composition using DEXA demonstrated that animals consuming the low viscosity supplements had significantly greater body fat than animals that consumed either the high viscosity supplement or no dietary supplement at all. These differences in body fat persisted for at least 3 months following the cessation of dietary supplements; during this 3-month period, animals previously exposed to the low viscosity supplement also gained significantly more weight than animals previously exposed to the high viscosity supplement. Taken together, the results suggest that consuming calories in low viscosity form may contribute to poor intake regulation over the short-term and to increased adiposity over the long term. When animals experience these diets as juveniles, these effects may persist into adulthood. PMID- 16243369 TI - Obesity: should treatments target visceral afferents? AB - The fact that obesity is a chronic disorder has traditionally focused experimental attention on the long-term controls of energy balance. Searches for therapeutic targets tend to concentrate on central integrative mechanisms and to largely ignore the visceral afferents and other peripheral mechanisms providing short-term controls of energy balance. Investigations of central mechanisms have yet to yield, however, any practical and effective treatments for correcting obesity. In this review, we survey some of the arguments for considering peripheral visceral afferent mechanisms as promising targets for future research on obesity. These arguments include (1) the observation that visceral afferents have the specializations, complexities, heterogeneities, and extensive distributions at key sites to provide exhaustive and dynamic feedback to control energy handling, (2) the fact that the most effective treatments yet developed for achieving long-term or permanent weight loss, namely gastroplasty and similar bariatric surgical procedures, clearly alter visceral afferent feedback from the gastrointestinal tract, and (3) experimental observations that suggest loss of visceral negative feedback can lead to overeating, positive energy balance, and obesity. Furthermore, even though excess adiposity is a disturbance in long-term energy regulation, it is instructive that obesity in the final analysis is developed, is maintained, and ultimately needs to be treated one meal at a time. When these considerations are taken in conjunction with concerns about side effects and risks that can be expected to accompany pharmacological therapies directed at central nervous system circuits, it would seem prudent to assess ways in which the feedback of visceral afferents might be enhanced or manipulated to support or synergize with other therapeutic strategies used in the management of excess energy intake. PMID- 16243370 TI - Prostate cancer cell type-specific involvement of the VDR and RXR in regulation of the human PTHrP gene via a negative VDRE. AB - Parathyroid hormone-related protein (PTHrP) increases the growth and osteolytic potential of prostate cancer cells, making it important to control PTHrP expression in these cells. We show that 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) and its non-hypercalcemic analog, EB1089, decrease PTHrP mRNA and cellular protein levels in the androgen-dependent human prostate cancer cell line LNCaP and its androgen-independent derivative, the C4-2 cell line. This effect is mediated via a negative Vitamin D response element (nVDREhPTHrP) within the human PTHrP gene and involves an interaction between nVDREhPTHrP and the Vitamin D receptor (VDR). The retinoid X receptor (RXR) is a frequent heterodimeric partner of the VDR. We show that RXRalpha forms part of the nuclear protein complex that interacts with nVDREhPTHrP along with the VDR in LNCaP and C4-2 cells. We also show that the RXR ligand, 9-cis-retinoic acid, downregulates PTHrP mRNA levels; this decrease is more pronounced in LNCaP than in C4-2 cells. In addition, 9-cis retinoic acid enhances the 1,25(OH)2D3-mediated downregulation of PTHrP expression in both cell lines; this effect also is more pronounced in LNCaP cells. Proliferation of LNCaP, but not C4-2, cells is decreased by 9-cis-retinoic acid. Promoter activity driven by nVDREhPTHrP cloned upstream of the SV40 promoter and transiently transfected into LNCaP and C4-2 cells is downregulated in response to 1,25(OH)2D3 and EB1089 in both cell lines. Co-treatment with these compounds and 9-cis-retinoic acid further decreases CAT activity in LNCaP, but not C4-2, cells. These results indicate that PTHrP gene expression is regulated by 1,25(OH)2D3 in a cell type-specific manner in prostate cancer cells. PMID- 16243371 TI - C-21 steroidal glycosides from the roots of Cynanchum chekiangense and their immunosuppressive activities. AB - Two new C21 steroidal glycosides, chekiangensosides A and B, were isolated from the roots of Cynanchum chekiangense, together with two known compounds. On the basis of chemical evidence and extensive spectroscopic methods, including one dimensional and two-dimensional NMR, the structures of two new compounds were identified as cynajapogenin A, 3-O-beta-D-glucopyranosyl-(1-->4)-beta-D cymaropyranosyl-(1-->4)-alpha-L-cymaropyranosyl-(1-->4)-beta-D-cymaropyranoside, and glaucogenin A, 3-O-beta-D-glucopyranosyl-(1-->4)-beta-D-cymaropyranosyl-(1- >4)-alpha-L-cymaropyranosyl-(1-->4)-beta-D-cymaropyranoside, respectively. The two known steroidal glycosides, and were revised. These isolated compounds were tested for their immunological activities in vitro against concanavalin A (Con A) and lipopolysaccharide (LPS)-induced proliferation of mice splenocytes. Compounds showed immunosuppressive activities in vitro in a dose-dependent manner. PMID- 16243372 TI - Competitive exclusion and limiting similarity: a unified theory. AB - Robustness of coexistence against changes of parameters is investigated in a model-independent manner by analyzing the feedback loop of population regulation. We define coexistence as a fixed point of the community dynamics with no population having zero size. It is demonstrated that the parameter range allowing coexistence shrinks and disappears when the Jacobian of the dynamics decreases to zero. A general notion of regulating factors/variables is introduced. For each population, its impact and sensitivity niches are defined as the differential impact on, and the differential sensitivity towards, the regulating variables, respectively. Either the similarity of the impact niches or the similarity of the sensitivity niches results in a small Jacobian and in a reduced likelihood of coexistence. For the case of a resource continuum, this result reduces to the usual "limited niche overlap" picture for both kinds of niche. As an extension of these ideas to the coexistence of infinitely many species, we demonstrate that Roughgarden's example for coexistence of a continuum of populations is structurally unstable. PMID- 16243373 TI - Comparison of shift estimation strategies in spectral elastography. AB - This paper compares the performance of various spectral shift estimators for use in spectral elastography, namely, the normalized cross-correlation (NCC), sum squared difference (SSD) and sum absolute difference (SAD). Simulation and experimental results demonstrate that the spectral SSD-based elastographic method exhibits no marked difference in performance compared to the more computationally costly NCC-based approach, which has conventionally been the preferred estimator in spectral elastography. The spectral SAD-based strain estimator, despite being computationally less burdening, failed to exhibit performance comparable to that of the NCC- and SSD-based techniques. Furthermore, though spectral subsample estimation techniques using a cosine-fit interpolation method outperformed that of the parabolic-fit method in terms of both reduced bias errors and standard deviations, the latter was analyzed in this study due to computational simplicity. The role of spectral density was evaluated without and with parabolic based subsample interpolation. Based on minimizing computational complexity, it is concluded that a (low density) spectral SSD strain estimator coupled with parabolic-based subsample estimation is the preferred choice for spectral elastography. PMID- 16243374 TI - Host cell killing by the West Nile Virus NS2B-NS3 proteolytic complex: NS3 alone is sufficient to recruit caspase-8-based apoptotic pathway. AB - The West Nile Virus (WNV) non-structural proteins 2B and 3 (NS2B-NS3) constitute the proteolytic complex that mediates the cleavage and processing of the viral polyprotein. NS3 recruits NS2B and NS5 proteins to direct protease and replication activities. In an effort to investigate the biology of the viral protease, we cloned cDNA encoding the NS2B-NS3 proteolytic complex from brain tissue of a WNV-infected dead crow, collected from the Lower Merion area (Merion strain). Expression of the NS2B-NS3 gene cassette induced apoptosis within 48 h of transfection. Electron microscopic analysis of NS2B-NS3-transfected cells revealed ultra-structural changes that are typical of apoptotic cells including membrane blebbing, nuclear disintegration and cytoplasmic vacuolations. The role of NS3 or NS2B in contributing to host cell apoptosis was examined. NS3 alone triggers the apoptotic pathways involving caspases-8 and -3. Experimental results from the use of caspase-specific inhibitors and caspase-8 siRNA demonstrated that the activation of caspase-8 was essential to initiate apoptotic signaling in NS3 expressing cells. Downstream of caspase-3 activation, we observed nuclear membrane ruptures and cleavage of the DNA-repair enzyme, PARP in NS3-expressing cells. Nuclear herniations due to NS3 expression were absent in the cells treated with a caspase-3 inhibitor. Expression of protease and helicase domains themselves was sufficient to trigger apoptosis generating insight into the apoptotic pathways triggered by NS3 from WNV. PMID- 16243376 TI - Surround-induced foveal afterimage pulsation: evidence for a long-range neural effect. AB - We demonstrate that an afterimage resulting from a strong foveal light flash can be made to pulsate by luminance modulation of a surrounding annulus as far as 8 deg away. Afterimage pulsation persists even if all artifacts due to pupil size, stray light and simultaneous contrast are ruled out. This suggests an origin by a long-range neural process acting from the remote surround. The effect is interpreted in terms of an adaptive gain control optimizing the responses of visual cells. PMID- 16243375 TI - Recovery of rod-mediated a-wave during light-adaptation in mGluR6-deficient mice. AB - The purpose of this study was to compare the a-waves of mGluR6-deficient mice (KO) to that of wild-type mice (WT), and to determine whether the light-adapted electroretinogram of the KO mice originate exclusively from cones. Dark-adapted a waves were recorded under the same conditions from both types of mice. With a 96 cd/m(2) background, the a-wave from both types of mice showed a rapid recovery over a 50-min period. The analysis of the a-waves in KO mice indicated that the recovery was determined mainly by the rod component. The light-adapted b-wave of WT mice showed no corresponding recovery. We conclude that rod contribution must be considered in the analyses of the light-adapted a-waves of KO mice. PMID- 16243377 TI - Removal of soluble COD by a biofilm formed on a membrane in a jet loop type membrane bioreactor. AB - The soluble chemical oxygen demand (COD) removal efficiency through a cake layer (biofilm) deposited on the surfaces of a membrane was investigated as a function of biofilm thickness in a jet loop type membrane bioreactor (JL-MBR). The mechanisms for the removal were investigated based on the microbial characteristics of the biofilm. Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) was used to identify the microbial community and a Specific oxygen uptake rate (SOUR) analysis was applied to determine the activities of microbial communities in a biofilm. Both the activities and community of microbial communities in the biofilm were similar to those found in a mixed liquor since, in JL-MBR all the substrates, dissolved oxygen, and nutrients are forced to flow through the biofilm, which differs from the biofilms grown on a non-permeable substratum in conventional biofilm process. The removal efficiency of soluble COD in a reactor through the active biofilm increased, reaching a constant value of approximately 92% despite the continuous increase in the thickness of active biofilm with the operation time. This might be attributed to (i) the presence of soluble COD that is not readily biodegraded, (ii) the presence of small and non-biodegradable organic molecules that could easily pass through the biofilm as well as the membrane, and (iii) too short a contact time of soluble solutes with the active microorganisms in the biofilm. PMID- 16243378 TI - Description, new reconstruction, comparative anatomy, and classification of the Sterkfontein Stw 53 cranium, with discussions about the taxonomy of other southern African early Homo remains. AB - Specimen Stw 53 was recovered in 1976 from Member 5 of the Sterkfontein Formation. Since its incomplete initial description and comparison, the partial cranium has figured prominently in discussions about the systematics of early Homo. Despite publication of a preliminary reconstruction in 1985, Stw 53 has yet to be compared comprehensively to other Plio-Pleistocene fossils or assessed systematically. In this paper, we report on a new reconstruction of this specimen and provide a detailed description and comparison of its morphology. Our reconstruction differs in important respects from the earlier one, especially in terms of neurocranial length, breadth, and height. However, given that Stw 53 exhibits extensive damage, these dimensions are most likely prone to much error in reconstruction. In areas of well-preserved bone, Stw 53 shares many cranial features with Homo habilis, and we propose retaining it within this species. We also consider the affinities of dental remains from Sterkfontein Member 5, along with those from Swartkrans and Drimolen previously assigned to Homo. We find evidence for sympatry of H. habilis and Australopithecus robustus and possibly Plio-Pleistocene Homo sapiens sensu lato in Sterkfontein Member 5. At Swartkrans and Drimolen, we find evidence of H. habilis. We also compare the morphologies of Stw 53 and SK 847 and find compelling evidence to assign the latter specimen to H. habilis, as has been proposed. PMID- 16243379 TI - Growth hormone administration to aged animals reduces disulfide glutathione levels in hippocampus. AB - Systemic growth hormone (GH) and insulin-like growth factor-1 (IGF-1), potent anabolic hormones, decrease with age. In humans and animal models, administration of growth hormone or IGF-1 to aged subjects improves learning and memory, suggesting that the age-related decline in cognitive performance results, in part, from peripheral GH/IGF-1 deficiency. However, the cellular mechanisms by which GH/IGF-1 effect cognitive function are unknown. We propose that the effects of these hormones may be mediated by increasing cellular redox potential resulting in reduced oxidative stress. Because the most abundant endogenous antioxidant is glutathione (GSH), we assessed GSH and disulfide glutathione (GSSH) levels in hippocampus and frontal cortex of young (4-month-old) and aged (30-month-old) male Fisher 344xBrown Norway rats treated with porcine growth hormone (200microg/animal, twice/daily) or vehicle. We report that hippocampal levels of GSSG increase with age (0.54+/-0.08 to 1.55+/-0.24nmolGSSG/mgprotein, p<0.05) and growth hormone treatment ameliorates both the age-related rise in GSSG (1.55+/-0.24 to 0.87+/-0.24nmolGSSG/mgprotein, p<0.05) and the decline in GSH/GSSG ratios. Analysis of GSSG reductase activity in aged animals indicated no effect of either age or growth hormone treatment (p=0.81). Although similar age related increases in GSSG and decreases in GSH/GSSG ratios were evident in frontal cortex, growth hormone had no effect. Subsequently, we assessed whether the effects of age and growth hormone treatment result from modulating trace metal accumulation. Thirteen metals were analyzed in hippocampus and frontal cortex by inductive coupled plasma mass spectrometry. Aluminum, copper, iron, manganese and zinc levels increased with age (p<0.05 each) but growth hormone replacement had no effect on metal accumulation. Our results indicate that growth hormone replacement attenuates the age-related increase in oxidative stress in hippocampus without effects on glutathione reductase or trace metal accumulation. We conclude that the age-related decline in circulating growth hormone and IGF-1 contribute to increased oxidative stress in hippocampus with age. PMID- 16243380 TI - Intraurban variations of PM10 air pollution in Christchurch, New Zealand: implications for epidemiological studies. AB - Epidemiological studies relating air pollution to health effects often utilise one or few central monitoring sites for estimating wider population exposures to outdoor particulate air pollution. These studies often assume that highly correlated particulate concentrations between intraurban sites equate to a uniform concentration field. Several recent studies have questioned the universal validity of this assumption, noting that in some cities, the uniformity assumption may lead to exposure misclassification in health studies. Few studies have compared central fixed site concentrations to intraurban population background sites using actual monitored data in cities with higher levels of pollution. This research examines daily concentration variations in particulate matter less than 10 microm in diameter (PM10) at the neighbourhood scale over two winter months in Christchurch, New Zealand, a city with high winter pollution concentrations. Daily concentrations of PM10 data were collected for two winter months at ten background monitoring sites within 9.3 km of the central fixed monitoring site typically used for estimating exposure in epidemiological studies. Results indicate that while the correlation between PM10 concentrations measured at the central monitoring site and most background sites is strong (r>0.76), absolute daily concentration differences between the central monitoring site and population background sites were substantial (mean 90th percentile absolute difference=17.6 microg m-3). In Christchurch, a central monitoring site does not therefore appear to accurately depict wider area population exposures to PM10. Local intraurban variations in particulates should be well understood before applying central monitoring site concentrations as proxies for population exposure in epidemiological studies. PMID- 16243382 TI - Transendothelial migration of ferric ion in FeCl3 injured murine common carotid artery. AB - INTRODUCTION/OBJECTIVES: Adventitial application of FeCl(3) causes endothelial injury, platelet aggregation, and a rapid onset of thrombus formation. The transmigration pathway of the ferric ion has not been definitively identified. Using a combination of TEM and X-ray elemental analysis, this study aims to elucidate the endothelial pathway of ferric ion migration in carotid artery. METHODS AND RESULTS: Vascular injury was induced by placing a Whatman #1 filter paper strip saturated with 10% FeCl(3) over the common carotid artery in male C57BL/6 mice for 3 min. After rinsing in saline, the mice were terminated at 10 or 30 min. The FeCL(3) exposed segments of the common carotid artery were dissected, and processed for TEM. Thrombus formation was observed in all cases. Endothelial and smooth muscle injuries were observed in segments of the vessel in direct contact with the oxidant. The endothelial injury ranged from minimal damage to total denudation. The basal endothelial surface adjacent to the internal elastic lamina showed accumulation of electron opaque vesicles. The membrane enclosed particles transmigrated across the endothelium and exocytosed into the lumen. The nature of the particles shown by STEM/EDS was rich in ferric ion. Elemental analysis also showed that some ferric oxide aggregates formed near the developing thrombus in the vascular lumen. CONCLUSION: Our results showed the ferric ion permeated the endothelial basal lamina before entering the arterial lumen via endocytic-exocytic pathway. This study provides an ultrastructural framework for future analysis of the adluminal and luminal injuries in this model. PMID- 16243381 TI - Mercury in different environmental compartments of the Pra River Basin, Ghana. AB - Artisanal gold mining (AGM) with metallic mercury has a long history in Ghana. It is believed to be over 2,000 years old. Today, AGM has escalated in a new dimension consuming about half of the country where gold lode deposits exist along riverbanks or rivers are alluvial-gold rich. The Pra River in southwestern Ghana is a site of on going application of metallic mercury in prospecting gold, and this paper examines mercury (Hg) contamination in the different environmental compartments in its watershed. Samples of water, sediment, soil and biota (i.e., human hair and fish) were collected from locations along the course of the river during the rainy and dry seasons of 2002 and 2003, respectively. Besides the obvious Hg point sources along the Pra and its tributaries, the obtained results show that Hg levels and speciation in the studied aquatic system are controlled by precipitation, which drives the hydrology and differences in flow regimes versus seasons. The seasonal difference in Hg speciation suggests that methyl mercury (MeHg) found in the aqueous phase and riverine sediments is likely of terrestrial origin where its production is favored during the rainy season by high soil water and organic matter content. The use of the enrichment factor (EF) for the assessment of sediment quality indicated moderate to severe contamination of surface sediments in the rainy season, while in the dry season, the EF index indicates nearly no pollution of surface sediments. Accordingly, most of the Hg introduced into this river system is likely transported to depositional downstream terminal basins (e.g. the river delta and the Gulf of Guinea). With regard to biota, Hg measured in hair in the dry period was higher than data obtained on samples collected during the wet period. This could be explained at least in part by the shift in diet as a result of abundance of fish in the local markets and the concurrent increase and more active fishing during the dry season. Mercury data obtained on a very limited number of fish samples collected during the dry period only are also presented. PMID- 16243383 TI - Endometrial volume and vascularity measurements by transvaginal three-dimensional ultrasonography and power Doppler angiography in stimulated and tumoral endometria: intraobserver reproducibility. AB - OBJECTIVES: To assess intraobserver reproducibility of the endometrial volume (EV) and 3D power Doppler indices (vascularization index, VI; flow index, FI; and vascularization flow index, VFI) of the endometrium and subendometrial area using three-dimensional power Doppler angiography (3D-PDA). METHODS: Twenty-five women on the hCG day after controlled ovarian stimulation and 15 patients presenting with uterine bleeding and suspicious endometrial thickening (10 endometrial cancers and 5 endometrial hyperplasias) were scanned. Eighty volume data sets were analyzed using the VOCAL imaging program. EV and VI, FI and VFI of the endometrium and subendometrium (5 mm shell) were manually calculated in the longitudinal and coronal planes with 15 degrees and 9 degrees rotation steps. Intraclass correlation coefficient (ICC) and 95% confidence intervals were used to assess reliability. RESULTS.: EV measurements were highly reproducible (ICC > or = 0.97) without significant differences between planes and rotation steps. Endometrial and subendometrial VI, FI, and VFI presented ICCs above 0.90 with the exception of the subendometrial FI (ICC > or = 0.80). There were no significant differences according to measurement plane and rotation step except for subendometrial VFI. Nevertheless, 3D power Doppler indices calculated in the coronal plane and 9 degrees rotation step obtained the highest ICC. ICCs for 3D PDA indices from the tumoral endometria were significantly higher than those calculated from the stimulated endometria. CONCLUSIONS: Endometrial volume and endometrial and subendometrial 3D power Doppler indices have an acceptable reproducibility, significantly higher in tumoral endometria. The reliability of measurements does not seem to be significantly influenced by the rotation plane and degrees of rotation. These results support that 3D-PDA and VOCAL are reliable methods to evaluate the physiological and pathological changes of the endometrium. PMID- 16243384 TI - Arsenic Trioxide inhibits the growth of human ovarian carcinoma cell line. AB - OBJECTIVE: To determine whether Arsenic Trioxide may be an effective treatment of human ovarian carcinoma, we examined if it induced apoptosis in human ovarian cancer cell line. METHODS: The human ovarian cell line SKOV3 was exposed to different concentrations of As2O3: 0 (control), 0.1 microM, 1 microM, 5 microM and 10 microM. The effect on cells proliferation and apoptotic parameters was examined. RESULTS: The most effective inhibitory Arsenic Trioxide concentrations were 5 microM and 10 microM, causing growth inhibition of 79% and 83%, respectively. The maximum inhibitory effect of Arsenic Trioxide on cellular proliferation was seen after 48 h. No morphological or molecular features of apoptosis have been observed. At the same time, there were no typical changes of cellular necrosis. CONCLUSIONS: The exact mechanism by which Arsenic Trioxide inhibits the growth of ovarian cancer cells is probably not by apoptosis. However, since As2O3 has been shown to inhibit ovarian cancer cellular growth, it should be further elucidated as a possible chemotherapeutic agent of ovarian cancer. PMID- 16243385 TI - Comparison of different transvaginal ovum pick-up protocols to optimise oocyte retrieval and embryo production over a 10-week period in cows. AB - The objective was to develop a simple and effective ovum pick-up (OPU) protocol for cows, optimised for oocyte harvest and subsequent in vitro embryo production (IVP). Five protocols differing in collection frequency, dominant follicle removal (DFR) and FSH stimulation were tested on groups of three cows each, over an interval of 10 consecutive weeks. Performance was evaluated on per OPU session, per week and pooled (3 cowsx10weeks) basis. Among the non-stimulated groups, on a per cow per session basis, once- or twice-weekly OPU had no effect on the mean (+/- S.E.M.) number of follicles aspirated, oocytes retrieved and blastocysts produced (0.6+/-0.8 and 0.7 +/- 0.7, respectively). However, DFR 72 h prior to OPU almost doubled blastocyst production (1.2 +/- 1.3). In stimulated groups, FSH treatment (80 mg IM and 120 mg SC) was given once weekly prior to OPU. Treatment with FSH, followed by twice-weekly OPU, failed to show any synergistic effect of FSH and increased aspiration frequency. When FSH was given 36 h after DFR, followed by OPU 48 h later, more (P < 0.05) follicles (16.0 +/- 5.0), oocytes (10.6 +/- 4.5) and embryos (2.1 +/- 1.2) were obtained during each session, but not on a weekly basis. Pooled results over 10 weeks showed an overall improved performance for the treatment groups with twice-weekly OPU sessions, due to double the number of OPU sessions performed. However, the protocol that consisted of DFR, FSH treatment and a subsequent single OPU per week, was the most productive and cost-effective, with potential commercial appeal. PMID- 16243386 TI - Characterization of calcium oscillation patterns in caprine oocytes induced by IVF or an activation technique used in nuclear transfer. AB - Routine activation of nuclear transfer (NT) eggs involves the application of a single intracellular calcium [Ca2+]i rise, stimulated by an electrical pulse, as opposed to [Ca2+]i oscillations, which is the natural mode of sperm-induced activation at fertilization in all mammalian species tested to date. It has yet to be shown that caprine oocytes exhibit an increase in calcium at fertilization in a manner similar to other mammals. The objective of the present study was to evaluate and characterize the ([Ca2+]i) oscillation patterns of caprine metaphase II (MII) oocytes during IVF and during an activation techniques used in nuclear transfer. Additionally, the effect of cytochalasin B (cyto B) in the NT process was evaluated for its impact on [Ca2+]i oscillations and subsequent embryo development. Mature in vitro and in vivo derived caprine oocytes were activated by 5 microM ionomycin, an electrical pulse(s), or IVF. The intracellular Ca2+ response was determined using the [Ca2+]i indicator Fura-2 dextran (Fura-2D). Ova treated with ionomycin or stimulated by an electrical pulse exhibited a single [Ca2+]i rise, whereas IVF-derived oocytes showed oscillations. IVF [Ca2+]i showed some variation, with 62% of in vitro matured oocytes exhibiting oscillations, whereas 8% of in vivo matured oocytes exhibited oscillations demonstrating a correlation between [Ca2+]i responses and maturation technique. Knowing the [Ca2+]i profile of activated eggs, one may be able to optimize the activation methodology used in a production nuclear transfer setting which could potentially improve development to term for NT embryos. PMID- 16243387 TI - Stem allomorphy in the Spanish mental lexicon: evidence from behavioral and ERP experiments. AB - This study presents results from a nonce-word elicited production task and a reading experiment using event-related brain potentials (ERPs) investigating finite forms of Spanish verbs which consist of marked stems and regular person and number agreement suffixes. The first experiment showed that unmarked stems are productively extended to nonce words, whereas marked stems generalize more restrictively to nonce words, based on lexical similarity to existing stem forms. The second experiment yielded a lexical ERP signature for stem violations and an ERP pattern signaling morpho-syntactic (rule-based) processing for suffix violations. We argue that stem allomorphy is lexically represented in the Spanish mental lexicon, with marked stems forming subnodes of structured lexical entries. PMID- 16243388 TI - Structure of lintnerized starch is related to X-ray diffraction pattern and susceptibility to acid and enzyme hydrolysis of starch granules. AB - Acid-resistant residues (lintnerized starches, Ls) were prepared from starches showing A-, B- and C- X-ray diffraction patterns. Ls retained the same X-ray crystalline type as their native counterparts with an improvement in diffraction intensity. Fluorophore-assisted capillary electrophoresis (FACE) study indicated that structural characteristics of Ls were associated with X-ray diffraction patterns. Double helices originated from linear chains with an approximate average degree of polymerisation (DP) 14, 16, and 15 would span the entire length of crystalline lamellae of A-, B-, and C-type starches, respectively. The proportion of singly branched materials (SB) with DP 25 protected in Ls was higher for A-type Ls (10-17%) than for B-type Ls (4-6%) and C-type Ls (8%). The structures of SB were similar in which branched chain (DP 13-15) was longer than main chain (DP 10-12). The structural characteristics of Ls are discussed in relation to acid and enzymatic degradations of starch granules. PMID- 16243390 TI - Maximum contractile filament movement per ATP used in muscle contraction is approximately 1.3 nm not 13 nm. PMID- 16243392 TI - Mechanisms of short crack growth at constant stress in bone. AB - This paper describes an experimental study of the growth of small (i.e. sub millimetre) cracks in samples of cortical bone subjected to a constant tensile stress. Slow, stable crack growth occurred at a rate and angle which were dependent on the orientation of the sample: tests were conducted with the loading axis both parallel and perpendicular to the longitudinal axis of the bone. All cracks showed intermittent growth in which periods of relatively rapid propagation alternated with periods of temporary crack arrest or relatively slow growth. In some cases crack arrest could be clearly linked to microstructural features such as osteons or Volkmann's canals, which acted as barriers to crack growth. Crack-opening displacement increased over time during the arrest periods. These observations suggest a mechanism for the growth of small cracks in bone at constant stress, involving microstructural barriers, time-dependent deformation of material near the crack tip and strain-controlled propagation. PMID- 16243391 TI - Design of modular non-viral gene therapy vectors. AB - Gene delivery has numerous potential applications both clinically and for basic science research. Non-viral vectors represent the long-term future of gene therapy and biomaterials are a critical component for the development of efficient delivery systems. Biomaterial development combined with fundamental studies of virus function and cellular processes will enable the molecular level design of modular vectors. Vectors are being developed based on cationic polymers or lipids that contain functional groups to mediate appropriate interactions with the extracellular environment or to interface with specific cellular processes. This review describes recent progress on the development of biomaterials for non viral vectors and highlights opportunities for future development. Ultimately, efficient vectors will expand the traditional applications of gene therapy within the clinic and may enable numerous other opportunities within diagnostics, biotechnology, and basic science research. PMID- 16243393 TI - The effect of structural alterations of PEG-fibrinogen hydrogel scaffolds on 3-D cellular morphology and cellular migration. AB - The need for alternative scaffolds in tissue engineering has motivated the establishment of advanced biomaterial technologies based on biosynthetic polymers. Networks of synthetic and biologic building blocks are created into a biomimetic environment for enhanced tissue compatibility with precise structural properties. The current investigation describes a unique biosynthetic hybrid scaffold comprised of synthetic polyethylene glycol (PEG) and endogenous fibrinogen precursor molecules. The PEGylated fibrinogen is cross-linked using photoinitation in the presence of cells to form a dense cellularized hydrogel network. The fibrin-like scaffold material maintains its biofunctionality through the fibrinogen backbone, while changes in the molecular architecture of the synthetic precursor are used to alter the nanostructrual properties of the scaffold, including mesh size and permeability. The structural properties of 6- and 10-kDa PEG-fibrinogen hydrogels are characterized by measuring the swelling properties and relating them to the degradation kinetics of the scaffold. Increased concentrations of the synthetic PEG are used to further alter the network structure of the PEG-fibrinogen hydrogel. Experiments using smooth muscle cells cultured inside the PEG-fibrinogen scaffold demonstrates a qualitative relationship between the molecular architecture of the matrix and the cellular morphology. A quantitative assessment of cell migration into the hydrogel network demonstrates a strong correlation between rate of cellular invasion and the network structure of the matrix. The ability to regulate cellular characteristics using structural modifications to the PEG-fibrinogen scaffold can be a valuable tool in tissue engineering and tissue regeneration. PMID- 16243394 TI - Biologically active lipid A antagonist embedded in a multilayered polyelectrolyte architecture. AB - Recently [Jessel N, Schwinte P, Donohue R, Lavalle P, Boulmedais F, Darcy R, et al. Pyridylamino-beta-cyclodextrin as a molecular chaperone for lipopolysaccharide embedded in a multilayered polyelectrolyte architecture. Adv Funct Mater 2004;14:963-9], we demonstrated the biological activity of a lipopolysaccharide from Escherichia coli incorporated into layer-by-layer films made of poly (l-lysine) and poly (l-glutamic acid) and containing a polycationic beta-cyclodextrin (CD) with chaperone properties. Here we develop innovative architectures containing a complex made of a charged beta-cyclodextrin and a lipid A antagonist (LAA) as potential systems for local endotoxin antagonistic activity. We examine the biological activity of these architectures. The CD-LAA complex adsorbed on top, or embedded into the polyelectrolyte films keeps its LPS antagonistic activity on both murine and human macrophages for at least 24h. PMID- 16243395 TI - High glucose-induced apoptosis through store-operated calcium entry and calcineurin in human umbilical vein endothelial cells. AB - Diabetes mellitus causes multiple cardiovascular complications. Previous studies have shown that prolonged exposure (96 h) of human umbilical vein endothelial cells (HUVECs) to hyperglycemia causes a significant increase in apoptosis. We report here that this increase in apoptosis is associated with an increase in Ca(2+) current (whole cell patch-clamp recorded) resulting from Ca(2+) entry mediated by store-operated channels (SOCs). The number of apoptotic cells after prolonged high glucose (HG, 30 mmol/L) exposure was significantly reduced in the presence of the SOC inhibitor 2-APB or of La(3+). A marked increase (approximately 80%) in Ca(2+)-dependent calcineurin (CN-A) phosphatase activity also occurred after prolonged HG exposure. Prolonged HG exposure-induced increase in CN-A activity was prevented by 2-APB, and selective CN-A phosphatase inhibition by FK506 or calmodulin inhibition by calmidazolium decreased HG induced apoptosis. Blocking hydrogen peroxide production using catalase or inhibiting the tyrosine kinase pp60(src) during prolonged exposure to HG, resulted in a marked decrease in apoptosis and was further associated with a significant reduction in CN-A phosphatase activity. The results demonstrate a significant role for Ca(2+) entry in HG-induced apoptosis in HUVECs, and suggest that this role is mediated via H(2)O(2) generation and the action of the Ca(2+) activated protein phosphatase calcineurin. PMID- 16243396 TI - Minimal residual core binding factor AMLs by real time quantitative PCR--initial response to chemotherapy predicts event free survival and close monitoring of peripheral blood unravels the kinetics of relapse. AB - Minimal residual disease (MRD) was measured by RQ-PCR in 11 AML1/ETO and 13 CBFbeta/MYH11 patients at diagnosis, after induction chemotherapy, and at all subsequent visits. Median detection limits were 1:50,000 and 1:10,000, respectively. In 64/103 samples MRD was detectable and highly correlated in PB and BM. In 38/103 samples, where MRD was only detectable in BM, median BM MRD was 3.5log lower than at diagnosis. Event free survival was significantly inferior in case of <2log reduction post-induction MRD. Persistent MRD was always followed by hematological relapse. Molecular progression rate in relapsing CBFbeta/MYH11 was surprisingly slow with a time lag to hematological relapse approaching 1 year. This direct comparison between the two subgroups of CBF AMLs delineates clear biological differences and corroborates the value of RQ-PCR. PMID- 16243397 TI - The central extended amygdala network as a proposed circuit underlying reward valuation. AB - The phenomenon of medial forebrain bundle self-stimulation offers a powerful model of reward-based behavior. In particular, it appears to activate a neural system whose natural function is to compute the survival value or utility of present stimuli and to help orchestrate responses toward those inputs. Although the anatomical identity of this system is as yet unknown, recent descriptions of anatomical macrosystems within the basal forebrain lead to the proposal that it may be largely contained within the central extended amygdala network. This paper reviews decades' worth of behavioral and neurophysiological investigations of brain stimulation reward that support or are at least consistent with this idea. The proposed network circuitry underlying self-stimulation is also placed into the larger context of basal forebrain function, specifically, the role of the ventral striatopallidum in linking motivation to behavior, the role of the amygdala in detecting motivationally significant inputs, and the role of the magnocellular complex in communicating reward information to cortical and hippocampal targets. PMID- 16243398 TI - Heavy metal concentrations in molluscs from the Senegal coast. AB - Bivalve molluscs from the Western coast of Senegal were sampled and measured for their cadmium, copper and zinc concentrations. This part of Africa has not been studied as regards heavy metal concentrations in the molluscs. The collected species are two small African bivalve molluscs, living in the sand: Cardita ajar and Dosinia isocardia; the African mussel Perna perna which is the only mussel of this genus in the Western coast of Africa and the oyster Crassostrea gasar, which lives in mangroves attached to the rhizophores, in the intertidal zone. C. gasar and C. ajar present higher cadmium concentrations, respectively 6.82+/-0.54 and 13.77+/-0.80 microg Cd/g (d.w.) than the two other species (D. isocardia: 3.88+/ 0.31 microg/g and P. perna 2.37+/-0.22 microg/g. Copper and zinc concentrations are in the range of the published values for C. gasar and P. perna collected elsewhere in Africa. The results suggest that cadmium may be present in high concentrations in Senegalese waters where upwellings occur. PMID- 16243399 TI - Short-term follow-up after implantation of a foldable iris-fixated intraocular lens in phakic eyes. AB - OBJECTIVE: To evaluate efficiency, predictability, stability, complications, and patient satisfaction after implantation of a foldable iris-fixated phakic intraocular lens (PIOL) for the correction of myopia. DESIGN: Prospective, nonrandomized, comparative (self-controlled) trial. PARTICIPANTS: Forty-one eyes of 22 myopic patients aged 18 to 56 years (mean, 36 years) with average sphere of -8.2+/-2.01 diopters (D; range, -12.25 to -3.75 D) and average preoperative cylinder of -0.90+/-0.62 D (range, -2.50 to 0.00 D) were enrolled in this prospective study. METHODS: All eyes underwent implantation of a foldable iris fixated PIOL with an optical zone of 6.0 mm. The follow-up was 6 months in all cases. Phakic intraocular lenses were available in powers ranging from -2.0 D to 12.0 D. MAIN OUTCOME MEASURES: The main parameters assessed were best spectacle corrected visual acuity (BSCVA), uncorrected visual acuity, refraction, endothelial cell count, intraocular pressure, slit-lamp biomicroscopy, and indirect ophthalmoscopy. RESULTS: At 6 months' follow-up, no eyes experienced a loss in BSCVA and 78% gained 1 or more lines of their preoperative BSCVA. Uncorrected visual acuity was significantly improved, with 82% of eyes reaching 20/25 or better. There was a significant reduction in spherical errors in all patients after surgery. Ninety-one percent of eyes were within +/-0.50 D of target refraction. A slight loss of endothelial cells (2.3 %) was observed 6 months after surgery. There were no intraoperative complications. In the postoperative follow-up, however, pigment precipitates were noted in 5 eyes of 4 patients. CONCLUSIONS: At short-term follow-up, the implantation of the foldable iris-fixated PIOL proved to be effective and predictable for the correction of myopia in phakic eyes. However, longer follow-up with larger numbers of patients is necessary to evaluate long-term complications. PMID- 16243400 TI - Cellular and molecular therapeutic modalities for arterial obstructive syndromes. AB - Arterial obstructive syndromes result in heart disease, stroke and limb loss, disability, and mortality. Currently available therapeutics for patients with these conditions are inadequate or fail in a significant number of patients. The development of novel therapies for severe coronary arterial disease (CAD), peripheral arterial disease (PAD), and cerebral vascular disease (CVD) is a major goal for modern medicine. Molecular and cell-based therapies for arterial obstructive syndromes have the potential to become clinically useful in the near future. Molecular therapy employs angiogenic proteins and genes in order to initiate the development of new blood vessels that by-pass an arterial occlusion. The induction of a collateral artery system is termed therapeutic angiogenesis or neovascularization. Proteins have been delivered either directly into the ischemic area or via a vector encoding an angiogenic gene. Both protein and gene therapies have been associated with promising preclinical and early phase human trial results in patients with PAD as well as CAD. However, to date, efficacy has not been demonstrated in placebo-controlled, large trails. Today's cell-based therapy is focused on stem cells (SCs) for the treatment of patients after acute myocardial infarction (AMI) or for patients with severe left ventricular dysfunction. Stem cells have shown to increase cardiac performance in uncontrolled, early phase human studies. This improvement is believed to have its origin in myogenesis and neovascularization. In the following review, we will cover current state of molecular- and cellular-based treatments for PAD and CAD that have reached the clinical arena. PMID- 16243401 TI - Carbohydrate-deficient transferrin parallels disease severity in anorexia nervosa. AB - Carbohydrate-deficient transferrin (CDT) is a commonly used biomarker for alcohol abuse; however, recent findings questioned its diagnostic value in catabolic subjects. We have thus investigated possible changes of CDT in patients suffering from eating disorders. Retrospectively, CDT values of patients with eating disorders were identified. Twenty-four non-alcoholic subjects could be found, in which CDT was determined at least once during stationary psychotherapy. Anorexia nervosa patients had pathological CDT concentrations in 57% of cases; conversely, bulimia nervosa patients had normal CDT. Patients with initially elevated CDT tended to be more seriously ill than those without. During therapy, the body mass index of anorexia nervosa patients normalized, paralleled by declining CDT. In anorexia nervosa patients, CDT is unsuitable as a marker of alcohol abuse, but it might serve as a parameter indicating prognosis and disease severity. However, case-control studies with larger samples are warranted. PMID- 16243402 TI - A mean red blood cell volume loss in tonsillectomy, adenoidectomy and adenotonsillectomy. AB - OBJECTIVE: To calculate a mean red blood cell volume (RBCV) loss per kilogram (kg) in adenoidectomy, tonsillectomy, and adeno-tonsillectomy. MATERIALS AND METHODS: Pre- and post-operative complete blood cell count, and pre-operative clotting studies of 144 patients were measured. Total blood volume (75 or 70 ml/kg), pre- and post-operative RBCV (hematocritxtotal blood volume), RBCV loss, per kg RBCV loss and per cent RBCV loss were calculated. RESULTS: There was significant differences between pre- and post-operative RBCV, Hb, and Htc values for tonsillectomy, adenoidectomy and A&T groups, respectively (p<0.001 for all groups). We found a mean RBCV loss of 33 ml (5.56% of total RBCV) for adenoidectomy, 128 ml (10.63% of RBCV) for tonsillectomy and 60 ml (10.71% of RBCV) for A&T. RBCV loss per kg was 1.57+/-1.29 for adenoidectomy, 2.96+/-1.91 for adeno-tonsillectomy, and 3.02+/-1.66 for tonsillectomy. CONCLUSION: According to us, for management of the patients, knowledge of a mean nature loss of RBCV per kg is important because bleeding seems to be unavoidable in the intra operative or maybe post-operative period in these operations. PMID- 16243403 TI - Thromboxane, prostacyclin and isoprostanes: therapeutic targets in atherogenesis. AB - Atherosclerosis is a chronic disease of the vasculature that is influenced by multiple factors that involve a complex interplay between some components of the blood and the arterial wall. Inflammation and oxidative stress have key roles in atherogenesis. The production of F2-isoprostanes (F2-IPs), thromboxane A2 (TxA2) and prostacyclin (PGI2) increases in atherosclerosis, and recent studies show that pharmacological modulation of their biosynthesis and biological activities are important therapeutic targets for managing atherosclerosis. In this review, we highlight recent breakthroughs in the roles of F2-IPs, TxA2 and PGI2 in atherogenesis, and identify pertinent therapeutic targets. PMID- 16243404 TI - Assessing economic and social pressure for the control of bovine viral diarrhoea virus. AB - The objective of this paper is to present a preliminary assessment of variation in the economic impact of bovine viral diarrhoea virus (BVDV) at dairy farm level between a sample of nations within the EU and hence assess differences in pressure to respond to this disease that may be impeding progress in control and hence restricting collective benefits from healthier livestock. We used a questionnaire to obtain national average values of key epidemiological and economic parameters for a typical dairy farm from BVDV experts in the countries concerned. These parameters were converted into assessments of economic impact using a computer simulation model. Uncontrolled output losses for a BVDV-naive herd with virus introduced in year 1 of a 10-year epidemic represented 22, 7, 8, 5, 8 and 20% of the BVDV-free annuity for the UK, Northern Portugal, Holland, Norway, Italy and Germany, respectively. Differences between countries will be widened by differences in the risk of acquiring BVDV. These will be much reduced in countries, such as Norway that have a national BVDV eradication programme. Farmers in such countries can therefore justify spending much less on maintaining BVDV-free status than BVDV-free farms in other countries. This result illustrates the paradox that in countries where BVDV prevalence is high, farmers have least to gain from unilateral BVDV eradication because of the high cost of maintaining freedom from the disease. We discuss this issue in the light of increasing recognition at international level of the importance of BVDV control. PMID- 16243405 TI - Analysis of the 1999-2000 highly pathogenic avian influenza (H7N1) epidemic in the main poultry-production area in northern Italy. AB - We evaluated the effects of risk factors and control policies following the highly pathogenic avian influenza (HPAI) epidemic that struck northern Italy's poultry industry in the winter of 1999-2000. The epidemic was caused by a type-A influenza virus of the H7N1 subtype, that originated from a low-pathogenic AI virus which spread among poultry farms in northeastern Italy in 1999 and eventually became virulent by mutation. Most infected premises (IP) were located in the regions of Lombardy and Veneto (382 out of 413, 92.5%), and the eradication measures provided for in the European legislation were enforced. In Veneto, where flock density was highest, infection-control was also accomplished by means of depopulation of susceptible flocks through a ban on restocking and pre-emptive slaughter of flocks that were in the vicinities of or that had dangerous contacts with IPs. In Lombardy, such control measures were applied to a lesser extent. Infection incidence rate (IR) was 2.6 cases per 1000 flocks per day in Lombardy and 1.1 in Veneto. After the implementation of infection-control measures, the at-risk population, the percentage of flocks < or =1.5 km from IPs, and the HPAI-IR underwent a greater reduction in Veneto than in Lombardy. Although the proximity (< or =1.5 km) to IPs in the temporal risk window (TRW) was a major risk factor for HPAI at the individual flock level, its effect at the population level (population-attributable fraction) did not exceed 31.3%. Viral transmission therefore also occurred among relatively distant flocks. Turkey flocks were characterised by greater IR of HPAI compared with other bird species such as layer hens, broilers, gamebirds, and waterfowl, even when located at distances >1.5 km from IPs. In Lombardy, IR for species other than turkeys was also relatively high. PMID- 16243406 TI - What influences participation in genetic carrier testing? Results from a discrete choice experiment. AB - This study explores factors that influence participation in genetic testing programs and the acceptance of multiple tests. Tay Sachs and cystic fibrosis are both genetically determined recessive disorders with differing severity, treatment availability, and prevalence in different population groups. We used a discrete choice experiment with a general community and an Ashkenazi Jewish sample; data were analysed using multinomial logit with random coefficients. Although Jewish respondents were more likely to be tested, both groups seem to be making very similar tradeoffs across attributes when they make genetic testing choices. PMID- 16243407 TI - Towards targeted mutagenesis and gene replacement in plants. AB - Advances in the development of biotechnological tools for plant gene disruption and repair have lagged behind the rapid progress made in whole-genome sequencing of many model and crop plant species. Plant DNA-repair machinery predominantly uses non-homologous end-joining (NHEJ), making the homologous recombination (HR) based methods, which have proved fruitful for gene targeting in non-plant systems, unsuitable for use in plant systems. Two recent reports describe successful targeted mutagenesis and gene targeting in Arabidopsis by either harnessing the plant NHEJ machinery using site-specific induction of double strand breaks (DSBs), or by activation of a HR pathway through overexpression of a yeast DNA recombination gene in transgenic plants. These reports provide a foundation from which new technologies for site-specific genome alterations in plant species can be developed. PMID- 16243408 TI - The role of biotechnology in art preservation. AB - Biotechnology has played a key role in medicine, agriculture and industry for over 30 years and has advanced our understanding of the biological sciences. Furthermore, the tools of biotechnology have a great and largely untapped potential for the preservation and restoration of our cultural heritage. It is possible that these tools are not often applied in this context because of the inherent separation of the worlds of art and science; however, it is encouraging to see that during the past six years important biotechnological applications to artwork preservation have emerged and advances in biotechnology predict further innovation. In this article we describe and reflect upon a unique example of a group of scientists and art restoration technicians working together to study and treat of a piece of colonial art, and review some of the new applications in biotechnology for the preservation of mankind's cultural heritage. We predict an expansion in this field and the further development of biotechnological techniques, which will open up new opportunities to both biologists and artwork preservers. PMID- 16243409 TI - Therapeutic ultrasound optimization for gene delivery: a key factor achieving nuclear DNA localization. AB - When applying therapeutic-ultrasound (TUS) for gene-delivery, there is a great need to understand the contribution of different parameters to the transfection process. The aim of this study is to optimize a wide range of parameters associated with the TUS system concurrent with parameters associated with the transfection, achieving high transfection level and efficiency (total number of cells), while localizing the DNA in the nucleus. Exposure of different cell-types (BHK, LNCaP, BCE) to TUS resulted in high gene expression (1200 fold) and efficiency (28%) with minimal loss in cell viability (<20%). The optimal transfection level and efficiency was achieved using TUS at 2 W/cm2 (0.159 MPa), 30% duty cycle (DC) for 30 min (1080 J/cm2), by placing the transducer above the cells. Long-term TUS application was found to overcome the rate-limiting step of this technology-driving DNA to the cell nucleus. The effect of cell density and DNA concentrations were studied. Increasing DNA concentration contributes to the increase in total gene expression, but not necessarily to transfection efficiency. Our findings support the feasibility of TUS to deliver genes to cells and contribute to the understanding of wide range of parameters that affect the capability of TUS to efficiently deliver genes. PMID- 16243410 TI - Age-related changes of vergence under natural viewing conditions. AB - Vergence eye movements were recorded with the scleral search-coil system in 32 healthy subjects (ages 19-73 years) to characterize the age-related effects on the dynamic parameters of vergence responses to step (transient components) and ramp or sinusoidal targets (sustained components) under natural viewing conditions. Transient vergence showed an age-related increase in latency and decreases in peak velocity and acceleration in the binocular stimulus condition but not in accommodative vergence. Sustained vergence showed no age-related effect in the binocular condition, but there was an age-related decrease in accommodative vergence steady-state velocity and an increase in latency. Age related changes of the transient and sustained components were very similar to those reported for saccades and smooth pursuit; they thus might support a distinction between a sustained and transient vergence system. Furthermore, such age-related effects have to be taken into account when assessing eye movement disorders in neurodegenerative and cerebrovascular diseases. PMID- 16243411 TI - Intranasal Protollin/F1-V vaccine elicits respiratory and serum antibody responses and protects mice against lethal aerosolized plague infection. AB - F1-V is a recombinant plague antigen comprising the capsular (F1) and virulence associated (V) proteins. Given intramuscularly with Alhydrogel, it protects mice against challenge, but is less effective in non-human primates against high-dose aerosolized Yersinia pestis challenge, perhaps because it fails to induce respiratory immunity. Intranasal immunization of mice with F1-V formulated with a Proteosome-based adjuvant (Protollin), elicited high titers of specific IgA in lungs whereas intranasal F1-V alone or intramuscular Alhydrogel-adsorbed F1-V did not. The Protollin-adjuvanted F1-V vaccine also induced high serum titers of specific IgG, comparable to those induced by intramuscular Alhydrogel-adsorbed F1 V. Mice immunized intranasally with Protollin-F1-V were 100% protected against aerosol challenge with 170 LD50 of Y. pestis and 80% against 255 LD50. PMID- 16243412 TI - Tracking mothers attitudes to childhood immunisation 1991-2001. AB - This report presents the findings from a series of 20 surveys carried out between 1991 and 2001. The main objectives of the research were to: This unique body of more than 15,000 interviews was conducted as part of a routine programme of research supporting the national immunisation programme in England. These surveys show that the public wants clarity, consistency, factual information and openness from those delivering immunisation services. PMID- 16243413 TI - Genetically engineered herpes simplex viruses that express IL-12 or GM-CSF as vaccine candidates. AB - We are using genetically modified, conditionally replicating herpes simplex virus (HSV) that express either interleukin (IL)-12 or granulocyte macrophage-colony stimulating factor (GM-CSF) as live, attenuated vaccine candidates for protection against HSV infection and/or disease. We report the following: (1) animals previously vaccinated with these candidate vaccines exhibited dose-dependent protection after intranasal, intraperitoneal or intracranial challenge with the highly virulent E377-MB wild-type HSV-1; (2) the IL-12 expressing virus (M002) consistently conferred protection at lower immunization doses than GM-CSF expressing virus (M004); (3) between 80 and 100% protection from E377-MB challenge was conferred after intramuscular immunization of mice with any of the three Deltagamma1 34.5 HSV, as opposed to 50% protection elicited after immunization with wild-type HSV-1 (F); and (4) latent virus was not detected at a higher rate in animals immunized and subsequently challenged with E377-MB than in immunized animals alone. These data suggest that conditionally replicating, cytokine-expressing HSV are able to elicit protective immune responses while retaining safety in an experimental murine model. PMID- 16243414 TI - Vertical distribution, migration rates, and model comparison of actinium in a semi-arid environment. AB - Vertical soil characterization and migration of radionuclides were investigated at four radioactively contaminated sites on Kirtland Air Force Base (KAFB), New Mexico to determine the vertical downward migration of radionuclides in a semi arid environment. The surface soils (0-15 cm) were intentionally contaminated with Brazilian sludge (containing (232)Thorium and other radionuclides) approximately 40 years ago, in order to simulate the conditions resulting from a nuclear weapons accident. Site grading consisted of manually raking or machine disking the sludge. The majority of the radioactivity was found in the top 15 cm of soil, with retention ranging from 69 to 88%. Two models, a compartment diffusion model and leach rate model, were evaluated to determine their capabilities and limitations in predicting radionuclide behavior. The migration rates of actinium were calculated with the diffusion compartment and the leach rate models for all sites, and ranged from 0.009 to 0.1 cm/yr increasing with depth. The migration rates calculated with the leach rate models were similar to those using the diffusion compartment model and did not increase with depth (0.045-0.076, 0.0 cm/yr). The research found that the physical and chemical properties governing transport processes of water and solutes in soil provide a valid radionuclide transport model. The evaluation also showed that the physical model has fewer limitations and may be more applicable to this environment. PMID- 16243415 TI - A study of Welsh mothers' experiences of postnatal depression. AB - OBJECTIVE: to examine the experiences of Welsh mothers diagnosed with postnatal depression and to question whether postnatal depression is socially determined. DESIGN: qualitative interviews using a semi-structured questionnaire to clarify mothers' thoughts and feelings within the postnatal period. SETTING: a semi-rural part of south-west Wales, previously dominated by the mining industry. PARTICIPANTS: ten mothers with one or more children participated in the study. Most of the mothers were from lower socio-economic groups. They were selected from 30 women diagnosed with postnatal depression. FINDINGS: mothers had little knowledge of the effects of postnatal depression before becoming pregnant, and were initially reluctant to confide or share their feelings. Economic pressures determined a second income and necessitated mothers returning to work. This left them with little quality time for their babies and family. The prime support networks, which in previous generations were grandmothers, were absent, and mothers relied on social services and voluntary support groups. IMPLICATIONS FOR PRACTICE: a greater understanding of the emotional and social effects of childbirth may help mothers to avoid feelings of isolation and the inability to cope. Discussing the issues of socio-emotional strain during pregnancy may help the mother to recognise the symptoms that identify postnatal depression, legitimise the condition and begin the recovery process. PMID- 16243416 TI - Appraising the quality of qualitative research. AB - In the process of undertaking a meta-synthesis of qualitative studies of free standing midwife-led units, the authors of this paper encountered a number of methodologically and epistemologically unresolved issues. One of these related to the assessment of the quality of qualitative research. In an iterative approach to scoping this issue, we identified eight existing checklists and summary frameworks. Some of these publications were opinion based, and some involved a synthesis of pre-existing frameworks. None of them provide a clear map of the criteria used in all their reviewed papers, and of the commonalities and differences between them. We critically review these frameworks and conclude that, although they are epistemologically and theoretically dense, they are excessively detailed for most uses. In order to reach a workable solution to the problem of the quality assessment of qualitative research, the findings from these frameworks and checklists were mapped together. Using a technique we have termed a 'redundancy approach' to eliminate non-essential criteria, we developed our own summary framework. The final synthesis was achieved through reflexive debate and discussion. Aspects of this discussion are detailed here. The synthesis is clearly rooted in a subjectivist epistemology, which views knowledge as constructed and hermeneutic in intent, encompassing individual, cultural and structural representations of reality. PMID- 16243417 TI - Management of tophaceous gout of the distal interphalangeal joint. AB - Surgical management of acute tophaceous gout of the distal interphalangeal joint is often associated with delayed wound healing. Aspiration through neighbouring uninvolved skin is a safe method of treating these tophi. For persistently symptomatic, unstable DIP joints, arthrodesis should be considered. PMID- 16243418 TI - Double-blind, comparative study of cyamemazine vs. bromazepam in the benzodiazepine withdrawal syndrome. AB - Cyamemazine is an anxiolytic antipsychotic, which reduces ethanol withdrawal symptoms. Here, we investigated if cyamemazine can be also effective as substitute drug to facilitate benzodiazepine withdrawal. A total of 168 patients treated with benzodiazepines for at least 3 months and with a <18 score in the Hamilton Anxiety Rating Scale (HARS) were included in the study. Previous benzodiazepine treatment was withdrawn, and patients were randomized to a 4-week treatment with cyamemazine (25-50 mg q.d.) or bromazepam (3-6 mg q.d.), followed by 2 weeks of placebo. The primary efficacy variable was the maximal anxiety rebound as measured with the HARS during the 42 days of treatment. No statistically significant differences between treatment groups were found for the extent or incidence of rebound anxiety. Considering all dropout patients as withdrawal failures, after 6 months of follow-up, 56/84 patients in the cyamemazine group (66.7%) and 55/84 patients in the bromazepam group (65.5%) were successfully withdrawn. 28 patients in the cyamemazine group and 18 in the bromazepam group had an adverse event, including anxiety, insomnia, dry mouth and somnolence. No extra-pyramidal symptoms were reported. In conclusion, cyamemazine was comparable to bromazepam in ensuring successful benzodiazepine withdrawal and in controlling the acute benzodiazepine withdrawal syndrome. Cyamemazine may be useful to facilitate benzodiazepine withdrawal in those patients where bromazepam substitution is not appropriate. PMID- 16243419 TI - Apoptotic effect of red wine polyphenols on human colon cancer SNU-C4 cells. AB - Polyphenols in fruits, soybean, vegetables, herbs, roots and leaves act as bioactive components related with prevention of cancer, heart diseases and diabetes. We investigated the apoptotic effects of polyphenols from red wine on human colon cancer cells SNU-C4 using 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) assay, 4,6-diamidino-2-phenylindole (DAPI) staining, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay, reverse transcription-polymerase chain reaction (RT-PCR) expressions of Bcl-2, Bax and Caspase-3 genes, and Caspase-3 enzyme activity. Polyphenols (100 microg/ml) increased the apoptosis of SNU-C4 cells with apparent apoptotic characteristics including morphological changes of chromatin condensation and apoptotic body formation from DAPI staining and TUNEL assay. Compared with untreated control group, polyphenols (100 microg/ml) reduced the expression of Bcl-2 whereas those of Bax and Caspase-3 were increased. The Caspase-3 activity in the polyphenols treated group was significantly increased compared to those in control group (P<0.05). These results suggest that polyphenols have a strong potential for development as an anti-colon cancer agent. PMID- 16243420 TI - A review of the toxicity of Melaleuca alternifolia (tea tree) oil. AB - The essential oil of Melaleuca alternifolia, also known as tea tree or melaleuca oil, is widely available and has been investigated as an alternative antimicrobial, anti-inflammatory and anti-cancer agent. While these properties are increasingly well characterised, relatively limited data are available on the safety and toxicity of the oil. Anecdotal evidence from almost 80 years of use suggests that the topical use of the oil is relatively safe, and that adverse events are minor, self-limiting and occasional. Published data indicate that TTO is toxic if ingested in higher doses and can also cause skin irritation at higher concentrations. Allergic reactions to TTO occur in predisposed individuals and may be due to the various oxidation products that are formed by exposure of the oil to light and/or air. Adverse reactions may be minimised by avoiding ingestion, applying only diluted oil topically and using oil that has been stored correctly. Data from individual components suggest that TTO has the potential to be developmentally toxic if ingested at higher doses, however, TTO and its components are not genotoxic. The limited ecotoxicity data available indicate that TTO is toxic to some insect species but more studies are required. PMID- 16243421 TI - [Malar valgisation in narrow face patients]. AB - Most of the "narrow faces" are secondary to a maxillo-malar hypoplasia consecutive to chronic nasal obstruction. Functional malar valgisation by transmalar osteotomy allows a simple and safe correction of this dysmorphic aspect. Interpositional bone graft and osteosynthesis by miniplate avoid mucous inflammation, maxillary sinusitis and/or relapse observed with use of biomaterials. Correction of "narrow faces" by malar osteotomy is natural and long term results are stable. Functional and esthetic rhinoplasty is often associated. Our clinical experience in 15 cases is reported. PMID- 16243422 TI - [Calciphylaxis: a severe but unrecognized complication in end-stage renal disease patients. A review of 2 cases]. AB - Calciphylaxis presents like subcutaneous lesions with livedo reticularis leading to necrotic and painful ulcers, predominantly in the lower limbs and the abdomen. They initially simulate dermohypodermitis. Biology reveals secondary hyperparathyroidism, phosphocalcic metabolism abnormalities and state of hypercoagulability. Histological signs are constant: calcifications in the media of small and sub-cutaneous arteries, intimal hyperplasia and intravascular thrombosis. This complication occurs in 4% of end-stage renal disease patients. Its prognostic is awful with a rate of mortality of 60% due to sepsis. Treatment is based upon the normalization of phosphocalcic rates and local debridement. PMID- 16243423 TI - [Scientific bases of fat transfer. Critical review of the literature]. AB - Fat grafting is an old technique. Since its first description in 1893, fat was used in a lot of indications. After more then a century of evolution, there is still no commonly accepted standard method of fat transfer by all the authors. On the contrary, many different techniques have been described in the literature with the only aim of reducing resorption of the transferred fat. Today the main inconvenient of fat grafting remains the partial resorption the transferred tissue. After a review of the literature of the various technique of fat transfer presented according to their chronology in the operative procedure, we have observed that the techniques developed by certain authors are the opposite of the techniques developed by others. This shows the lack of rigorous work concerning the fat transfer. Whether they are experimental or clinical, in most of these studies, we can find at least one skew which could have influence the results and the conclusion of these studies. A critical revue of the literature offers the opportunity to reconsider the actual bases of fat transfer. The use of an objective way of quantifying the volumes in clinical practice could permit the comparison of different studies in the future. PMID- 16243424 TI - Antioxidant activity of caffeic acid (3,4-dihydroxycinnamic acid). AB - Caffeic acid (3,4-dihydroxycinnamic acid) is among the major hydroxycinnamic acids present in wine; sinapic acid, which is a potent antioxidant. It has also been identified as one of the active antioxidant. In the present study, the antioxidant properties of the caffeic acid were evaluated by using different in vitro antioxidant assays such as 2-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) radical scavenging, 1,1-diphenyl-2-picryl-hydrazyl free radical (DPPH) scavenging, total antioxidant activity by ferric thiocyanate method, total reductive capability using the potassium ferricyanide reduction method, superoxide anion radical scavenging and metal chelating activities. alpha Tocopherol, trolox, a water-soluble analogue of tocopherol, butylated hydroxyanisole (BHA), and butylated hydroxytoluene (BHT) were used as the reference antioxidant compounds. At the concentrations of 10 and 30 microg/mL, caffeic acid showed 68.2 and 75.8% inhibition on lipid peroxidation of linoleic acid emulsion, respectively. On the other hand, 20 microg/mL of standard antioxidant such as BHA, BHT, alpha-tocopherol and trolox indicated an inhibition of 74.4, 71.2, 54.7 and 20.1% on peroxidation of linoleic acid emulsion, respectively. In addition, caffeic acid is an effective ABTS(+) scavenging, DPPH scavenging, superoxide anion radical scavenging, total reducing power and metal chelating on ferrous ions activities. PMID- 16243425 TI - The role of Nurr1 in the development of dopaminergic neurons and Parkinson's disease. AB - Nurr1, a transcription factor belonging to the orphan nuclear receptor superfamily, is critical in the development and maintenance of the dopaminergic system and as such it may have role in the pathogenesis of Parkinson' disease (PD). Human Nurr1 gene has been mapped to chromosome 2q22-23 and Nurr1 protein is predominantly expressed in central dopaminergic neurons. Nurr1 interacts with other factors critical for the survival of mensencephalic dopaminergic neurons and it appears to regulate the expression of tyrosine hydroxylase (TH), dopamine transporter (DAT), vesicular monoamine transporter 2 (VMAT2), and l-aromatic amino acid decarboxylase (AADC), all of which are important in the synthesis and storage of dopamine. Experimental studies in Nurr1 knock-out mice indicate that Nurr1 deficiency results in impaired dopaminergic function and increased vulnerability of those midbrain dopaminergic neurons that degenerate in PD. Decreased Nurr1 expression is found in the autopsied PD midbrains, particularly in neurons containing Lewy bodies, as well as in peripheral lymphocytes of patients with parkinsonian disorders. Several variants in Nurr1 gene have been reported in association with PD. All these studies suggest that Nurr1 is not only essential in the development of mensencephalic dopaminergic neurons and maintenance of their functions, but it may also play a role in the pathogenesis of PD. PMID- 16243426 TI - Caesarean before labour between 34 and 37 weeks: what are the risk factors of severe neonatal respiratory distress? AB - OBJECTIVE: To assess the frequency of severe neonatal respiratory distress and identify its risk factors in caesarean deliveries before labour between 34 and 37 weeks' gestation. STUDY DESIGN: Retrospective study of children born by caesarean delivery before labour between 34 and 37 weeks, between 1999 and 2003 in a level 3 maternity unit. The frequencies of severe and mild neonatal respiratory distress were calculated. Univariate and multivariate analyses studied the factors potentially associated with severe respiratory distress: gestational age, type of pregnancy (singleton or multiple), condition of membranes, maternal diabetes, indication for caesarean, antenatal corticosteroid therapy, intrauterine growth retardation, infant's sex and birth weight. RESULTS: The 189 study subjects included 107 singletons and 82 twins: 28% required intensive care for severe respiratory distress and 30.2% developed mild respiratory distress. Gestational age was a significant risk factor (p = 0.01), especially before 36 weeks (adjusted OR = 2.1; 95% CI: 1.0-4.4). The multivariate analysis indicated that singleton pregnancies (adjusted OR = 3.2; 95% CI: 1.5-6.7) and caesareans for fetal indications (adjusted OR = 2.7; 95% CI: 1.2-5.7) are also risk factors and that premature rupture of the membranes is a "protective" factor against respiratory complications (adjusted OR = 0.2; 95% CI: 0.1-0.8). CONCLUSION: More than a quarter of the infants delivered by caesarean before labour between 34 and 37 weeks' gestation in our level 3 maternity unit had severe respiratory distress. Although our population may not be typical of the general population, this finding and the risk factors associated with it should be taken into account in determining the best time and place for delivery of each patient. PMID- 16243427 TI - Oral N-acetylcysteine administration does not stabilise the process of established severe preeclampsia. AB - OBJECTIVE: To stabilise the disease process in women with early onset severe preeclampsia and/or HELLP syndrome by enhancing maternal antioxidants effects of glutathione. STUDY DESIGN: In a randomised, double-blind, placebo-controlled trial, women with severe preeclampsia and/or HELLP syndrome received oral N acetylcysteine. Primary outcome measures were disease stabilisation expressed as treatment-to-delivery interval and biochemical assessment of glutathione and parameters of oxidative stress. Secondary outcome measures were maternal complications, rate of caesarean section, stay at intensive care unit, postpartum hospital stay and neonatal morbidity and mortality. Analyses were done by intention-to-treat using Wilcoxon's two-sample test and regression analysis. RESULTS: Median treatment-to-delivery interval was not significantly different between the N-acetylcysteine and placebo group. The whole blood and plasma levels of glutathione and other thiols were not affected by N-acetylcysteine administration, except for plasma homocysteine concentrations, which were lower in the N-acetylcysteine group. There were no differences found in maternal nor neonatal secondary outcome measures between both groups. CONCLUSION: Oral N acetylcysteine administration does not stabilise the disease process of early onset severe preeclampsia and/or HELLP syndrome. PMID- 16243428 TI - Nanoparticle formation from poly(acrylic acid) and oppositely charged peptides. AB - Cationic peptides self assemble upon interacting with sodium salt of oppositely charged polymer, poly(acrylic acid), PAA, giving rise to water-soluble nanoparticles at very low concentration (0.1 mM of PAA). The morphology of these kinds of nanoparticles is mainly governed by the composition of the complexes, which can be expressed as Z+/-, i.e., the ratio of positively charged units to the concentration of anionic units of the polymers present in the system. In the present study, at lower Z+/-, the particles are elongated in shape but adopt spherical shape of 75-100 nm in diameter at higher Z+/- values. We propose that the nanoparticles containing cationic peptides obtained by this methodology can serve as delivery system to enhance the antinociception effect of the chimeric peptide with previously administered doses. PMID- 16243429 TI - Perturbing effects of carvedilol on a model membrane system: role of lipophilicity and chemical structure. AB - Carvedilol, a beta-adrenergic blocker used to treat cardiovascular diseases, protects cell membranes from lipid peroxidative damage. Previous studies suggested the drug resides in a non-polar environment and partitions into cell membranes, perturbing their fluidity. Here differential scanning calorimetry (DSC) and fluorescence spectroscopy were applied to further investigate interactions of carvedilol with a liposome model. Results indicate the association is relatively unaffected by pH or temperature, but could be sensitive to liposome composition. The drug's carbazole group plays the dominant role in bilayer perturbation. Compared with other beta-blockers examined, carvedilol produced the strongest liposome DSC perturbation. Locations of carbazole and carvedilol in the liposome were determined using depth-dependent fluorescent probes. Both compounds are situated in the middle of the bilayer, consistent with strong hydrophobic interactions. This combination of high lipophilicity and specific chemical structure appear required for carvedilol's novel antioxidant activity, and may enhance cardioprotection. PMID- 16243431 TI - High frequency of mutation of epidermal growth factor receptor in lung adenocarcinoma in Thailand. AB - Recent reports have suggested influences of racial difference on the frequency of mutation of EGFR in lung cancer. We therefore sought to characterize the frequency and pattern of mutation of EGFR in lung adenocarcinoma in Thai patients. Overall, EGFR catalytic domain mutations were detected in 35/61 (57.4%). We found 29/60 (48.3%) of exon 19 deletions, 5/54 (9.3%) of exon 21 point mutations, and 1/54 (1.9%) of double-mutation of both exons. The presence of these mutations was significantly associated with non-smoking habit. In summary, we report a strikingly high prevalence of mutation of EGFR in Thai lung adenocarcinoma, which may explain the high response rate to the treatment with TKI among Asian populations. PMID- 16243430 TI - Effects of COX-2 inhibitor on growth of human gastric cancer cells and its relation to hepatocyte growth factor. AB - It is known that hepatocyte growth factor binding to its receptor regulates gastric cancer progression and metastasis. HGF was found to up-regulate the expression of cyclooxygenase-2 gene and increases prostaglandin (PG) synthesis in gastric mucosa cells. Overexpression of COX-2 and increased PG secretion have also been found to be involved in the regulation of growth and metastasis of gastric cancer. Results from this study showed that c-Met and COX-2 are expressed in 28 cases (93.3%) and 16 cases (53.3%) of 30 human gastric cancer tissues, respectively. Expressions of c-Met positively correlated with that of COX-2 (r=0.41; P=0.024). Using in vivo and in vitro models to further examine the interaction between c-MET and COX-2, we found that HGF stimulated the growth of SC-M1 cells in a dose-dependent manner. COX-2-specific inhibitor-NS398 inhibited the growth of human gastric cancer SC-M1 cells as well as HGF stimulated the growth of SC-M1 cells in a dose-dependent manner. HGF treatment of SC-M1 cells increased the secretion of PGE2 and this stimulation was blocked by NS398. In vivo SC-M1 tumor model showed that HGF stimulated the tumor growth and NS398 retarded the tumor growth. These results suggest that COX-2-specific inhibitors may play some role on the therapy of gastric cancer patients with high serum HGF level and overexpression of c-Met in tumor. PMID- 16243432 TI - Global kinetic model: a case study on the N-oxidation of alkylpyridines. AB - The homogeneous catalytic N-oxidation of two picolines and two lutidines by hydrogen peroxide has been studied calorimetrically using a heat flow and power compensation SIMULAR reaction calorimeter. The objective of this work was to extend a previously developed kinetic model [J. Sempere, R. Nomen, J.L. Rodriguez, M. Papadaki, Modelling of the reaction of 2-methylpyridine using hydrogen peroxide and a complex metal catalyst, Chem. Eng. Process. 37 (1998) 33 46] for 2-methylpyridine to more reactions in the same family. The kinetic model is in good agreement with our experimental data on beta-picoline. 3,5-Lutidine is in adequately good agreement with the model. However, the formation of two phases during the course of the reaction imposes the need for modifications to allow for mass transfer considerations. The N-oxidation of 2,6-lutidine is controlled by the addition or availability of hydrogen peroxide. A different reactor design is necessary for operation at the kinetic regime. High temperatures and catalyst concentrations enhance the selectivity towards N-oxidation of all alkylpyridines studied. The power evolution of all reactions has similar profiles thus indicating that similar kinetics are followed. The study of the N-oxidation of this family of compounds indicates that it is possible to design a process where hydrogen peroxide decomposition can be practically totally suppressed. PMID- 16243433 TI - Enhanced reduction of perchlorate by elemental iron at elevated temperatures. AB - Kinetics of perchlorate reduction by elemental iron was examined at elevated temperatures using microwave heating and conventional block heating. It was hypothesized that increasing the solution temperature may accelerate the reduction of perchlorate by overcoming the high activation energy barrier. Results from microwave heating study showed that 98% of aqueous perchlorate was removed in 1 h at 200 degrees C. Similar results observed in control experiments with a block heater indicated that the enhancement in the extent and rate of perchlorate removal by elemental iron was mostly due to heat energy at high temperature. The rapid and complete reduction of perchlorate by elemental iron at elevated temperatures suggests that iron reduction process at elevated temperature may be an option to consider for complete removal of perchlorate from industrial discharges. PMID- 16243434 TI - Contribution to the study of electrocoagulation mechanism in basic textile effluent. AB - Electrocoagulation method with iron electrode is used to treat the industrial textile wastewater in batch reactor. The effects of operating parameters such as time and potential electrolysis on the decolourization and COD removal efficiency have been investigated. The results indicate that electrocoagulation is very efficient and able to achieve 100% colour and 84% COD removal in 3 min at potential 600 mV. The effluent wastewater is very clear and its quality exceed the direct discharge standard. Furthermore, the mechanism of electrocoagulation is elucidated by zeta potential measurement. PMID- 16243435 TI - Analgesic effects of capsazepine and resiniferatoxin on bone cancer pain in mice. AB - In the present paper, we describe the analgesic effects induced by the transient receptor potential vanilloid type 1 (TRPV1) antagonist, capsazepine, and the TRPV1 agonist, resiniferatoxin, on the thermal hyperalgesia induced by the presence of a tibial osteosarcoma or an inflammatory process in mice. The administration of capsazepine abolished the osteosarcoma-induced hyperalgesia at a dose range (3-10 mg/kg; s.c.) ineffective to inhibit the hyperalgesia elicited by the intraplantar administration of complete Freund's adjuvant (CFA). In contrast, the administration of resiniferatoxin (0.01-0.1 mg/kg; s.c.) inhibited both the osteosarcoma- and the CFA-induced hyperalgesia. Remarkably, a single dose of resiniferatoxin abolished the osteosarcoma-induced hyperalgesia for several days and completely prevented the instauration of thermal hyperalgesia when administered at the initial stages of osteosarcoma development. The potential of drugs acting through TRPV1 for the management of some types of bone cancer pain is proposed. PMID- 16243436 TI - Brain ischemia/reperfusion-induced expression of DP5 and its interaction with Bcl 2, thus freeing Bax from Bcl-2/Bax dimmers are mediated by c-Jun N-terminal kinase (JNK) pathway. AB - Our previous studies and those of others have strongly suggested that c-Jun N terminal kinase (JNK) signaling pathway plays a critical role in ischemic brain injury. But the downstream mechanism that accounts for the proapoptotic actions of JNK during cerebral ischemia/reperfusion still remains to be investigated in detail. DP5, one of the mammalian BH3-only proteins, was cloned as a neuronal apoptosis-inducing gene. In this study, we examined the changes of protein level of DP5 and its interaction with Bcl-2 family members in a rat model of global ischemia and reperfusion by immunoprecipitation and immunoblotting; furthermore, we investigated the effect of activated JNK on DP5-signaling pathway. We show here that DP5 was induced and interacted with Bcl-2 but not Bax in hippocampal CA1 6 h to 3 days after ischemia, while the interaction of Bcl-2 with Bax decreased. Systemic administration of SP600125, a small molecule JNK-specific inhibitor, diminished the induction of DP5 and its interaction with Bcl-2 after 2 days of ischemia. At the same time, SP600125 increased the interaction of Bax with Bcl-2 after 2 days of reperfusion. Thus, these results indicate that brain ischemia/reperfusion-induced activation of DP5 signaling pathway is mediated by JNK in postischemic rat hippocampal CA1. PMID- 16243438 TI - High-quality evidence in oncology from 14 April to 12 May 2005: a summary. PMID- 16243437 TI - Gain-of-function screen identifies a role of the Sec61alpha translocon in Drosophila postmitotic neurotoxicity. AB - To elucidate the intrinsic mechanisms of neurotoxicity induction, including those underlying neural cell death and neurodegeneration, we developed a gain-of function screen for gene products causing neural cell loss. To identify novel genes with a cell-death-related function in neurons, we screened 4,964 Drosophila GS lines, in which one or two genes from much of the Drosophila genome can be overexpressed. Approximately 0.68% of the GS lines produced phenotypes involving a loss of postmitotic neurons. Of these, we identified and characterized the endd2 gene, which encodes the Drosophila ortholog of Sec61alpha (DSec61alpha), an endoplasmic reticulum protein with protein translocation activity. Ectopic expression of DSec61alpha caused neural cell death accompanied by the accumulation of ubiquitinated proteins, which was mediated by DSec61alpha's translocon activity. This supported our previous observation that the DSec61alpha translocon contributes to expanded polyglutamine-mediated neuronal toxicity, which is also associated with ubiquitinated protein accumulation. These data suggest that the translocon may be a novel component of neural cell death and degeneration pathways. Our approach can be used to identify potential neurotoxic factors within the whole genome, which will increase our understanding of the molecular mechanisms of various types of cell death, including those associated with human neurodegenerative diseases. PMID- 16243439 TI - Follow-up difficulty: correlates and relationship with outcome in heroin dependence treatment in the NEPOD study. AB - Data collected from 317 heroin users who participated in four studies that were included in the Australian National Evaluation of Pharmacotherapies for Opioid Dependence were analysed to examine predictors of follow-up difficulty and whether follow-up difficulty was related to heroin use outcomes. Participants who were no longer receiving treatment were more difficult to contact and more likely to be lost to follow-up. Participants treated in general practice settings were also more difficult to contact and more likely to be lost to follow-up than participants treated at specialist clinics. Contact difficulty among followed-up participants (either in or out of treatment) was unrelated to heroin use outcomes. The 21% of participants who were followed-up with just one contact attempt reported 20.0 heroin-free days in the previous month, increasing only slightly to 20.9 based on the 70% of participants eventually contacted after up to 20 attempts. The study examined three methods for imputing missing heroin use outcome data and concluded that imputation of missing outcome data by inserting corresponding baseline data may be too conservative. PMID- 16243440 TI - Intravenous amifostine during chemoradiotherapy for head-and-neck cancer: a randomized placebo-controlled phase III study. AB - PURPOSE: Clinical trials demonstrated the efficacy and safety of intravenous (i.v.) or subcutaneous (s.c.) amifostine for reducing xerostomia and mucositis after radiotherapy or radiochemotherapy for head-and-neck cancer. This randomized, double-blinded, placebo-controlled, phase III study evaluated the efficacy and safety of i.v. amifostine during radiochemotherapy for head-and-neck cancer. METHODS AND MATERIALS: Patients from European and American study centers received i.v. amifostine 300 mg/m2 (n = 67) or placebo (n = 65) before carboplatin 70 mg/m2 and radiotherapy on Days 1 to 5 and 21 to 25, and i.v. amifostine 200 mg/m2 or placebo before radiotherapy on other days. RESULTS: Toxicity incidences were (amifostine, placebo, p value): Grade 2 or higher acute xerostomia (39%, 34%, 0.715), Grade 3 or higher acute mucositis (39%, 22%, 0.055), Grade 2 or higher late xerostomia (37%, 24%, 0.235), and Grade 3 or higher treatment-related adverse events (42%, 20%, 0.008). One-year rates of locoregional failure, progression-free survival, and overall survival were not significantly different between treatments. CONCLUSIONS: The used amifostine doses were not able to reduce the toxicity of simultaneous radiochemotherapy for head-and-neck cancer. The safety of amifostine and the lack of tumor protection were consistent with previous studies. PMID- 16243441 TI - Comparison of day 0 and day 14 dosimetry for permanent prostate implants using stranded seeds. AB - PURPOSE: To determine, using MRI-based dosimetry (Day 0 and Day 14), whether clinically significant changes in the dose to the prostate and critical adjacent structures occur between Day 0 and 14, and to determine to what degree any changes in dosimetry are due to swelling or its resolution. METHODS AND MATERIALS: A total of 28 patients with a permanent prostate implant using 125I rapid strands were evaluated at Days 0 and 14 by CT/MRI fusion. The minimal dose received by 90% of the target volume (prostate D90), percentage of volume receiving 100% of prescribed minimal peripheral dose (prostate V100), external sphincter D90, and 4-cm3 rectal volume dose were calculated. An acceptable prostate D90 was defined as D90 >90% of prescription dose. Prostate volume changes were calculated and correlated with any dosimetry change. A paradoxic dosimetric result was defined as an improvement in D90, despite increased swelling; a decrease in D90, despite decreased swelling; or a large change in D90 (>30 Gy) in the absence of swelling. RESULTS: The D90 changed in 27 of 28 patients between Days 0 and 14. No relationship was found between a change in prostate volume and the change in D90 (R2 = 0.01). A paradoxic dosimetric result was noted in 11 of 28 patients. The rectal dose increased in 23 of 28 patients, with a >30-Gy change in 6. The external sphincter D90 increased in 19 of 28, with a >50-Gy increase in 6. CONCLUSION: The dose to the prostate changed between Days 0 and 14 in most patients, resulting in a change in clinical status (acceptable or unacceptable) in 12 of 28 patients. Profound increases in normal tissue doses may make dose and toxicity correlations using Day 0 dosimetry difficult. No relationship was found between the prostate volume change and D90 change, and, in 11 patients, a paradoxic dosimetric result was noted. A differential z-axis shift of stranded seeds vs. prostate had a greater impact on final dosimetry and dose to critical adjacent tissues than did prostate swelling. These findings challenge the model that swelling is the principal cause of dosimetric changes after implantation. Stranded seeds may have contributed to this outcome. On the basis of these findings, a change in technique to avoid placement of stranded seeds inferior to the prostate apex has been adopted. These results may not apply to implants using single seeds within the prostate. PMID- 16243443 TI - Prostate position variability and dose-volume histograms in radiotherapy for prostate cancer with full and empty bladder. AB - PURPOSE: To evaluate prostate position variability and dose-volume histograms in prostate radiotherapy with full bladder (FB) and empty bladder (EB). METHODS AND MATERIALS: Thirty patients underwent planning computed tomography scans in a supine position with FB and EB before and after 4 and 8 weeks of radiation therapy. The scans were matched by alignment of pelvic bones. Displacements of the prostate/seminal vesicle organ borders and center of mass were determined. Treatment plans (FB vs. EB) were compared. RESULTS: Compared with the primary scan, FB volume varied more than EB volume (standard deviation, 106 cm3 vs. 47 cm3), but the prostate/seminal vesicle center of mass position variability was the same (> 3 mm deviation in right-left, anterior-posterior, and superior inferior directions in 0, 41%, and 33%, respectively, with FB vs. 0, 44%, and 33% with EB). The bladder volume treated with 90% of the prescription dose was significantly larger with EB (39% +/- 14% vs. 22% +/- 10%; p < 0.01). Bowel loops received > or = 90% of prescription dose in 37% (3% with FB; p < 0.01). CONCLUSION: Despite the larger variability of bladder filling, prostate position stability was the same with FB compared with EB. An increased amount of bladder volume in the high-dose region and a higher dose to bowel loops result from treatment plans with EB. PMID- 16243442 TI - Prospective evaluation of concurrent paclitaxel and radiation therapy after adjuvant doxorubicin and cyclophosphamide chemotherapy for Stage II or III breast cancer. AB - PURPOSE: To evaluate the safety and feasibility of concurrent radiation therapy and paclitaxel-based adjuvant chemotherapy, given either weekly or every 3 weeks, after adjuvant doxorubicin and cyclophosphamide (AC). METHODS AND MATERIALS: After definitive breast surgery and AC chemotherapy, 40 patients with operable Stage II or III breast cancer received protocol-based treatment with concurrent paclitaxel and radiation therapy. Paclitaxel was evaluated on 2 schedules, with treatment given either weeklyx12 weeks (60 mg/m2), or every 3 weeksx4 cycles (135 175 mg/m2). Radiation fields and schedules were determined by the patient's surgery and pathology. The tolerability of concurrent therapy was evaluated in cohorts of 8 patients as a phase I study. RESULTS: Weekly paclitaxel treatment at 60 mg/m2 per week with concurrent radiation led to dose-limiting toxicity in 4 of 16 patients (25%), including 3 who developed pneumonitis (either Grade 2 [1 patient] or Grade 3 [2 patients]) requiring steroids. Efforts to eliminate this toxicity in combination with weekly paclitaxel through treatment scheduling and CT-based radiotherapy simulation were not successful. By contrast, dose-limiting toxicity was not encountered among patients receiving concurrent radiation with paclitaxel given every 3 weeks at 135-175 mg/m2. However, Grade 2 radiation pneumonitis not requiring steroid therapy was seen in 2 of 24 patients (8%) treated in such a fashion. Excessive radiation dermatitis was not observed with either paclitaxel schedule. CONCLUSIONS: Concurrent treatment with weekly paclitaxel and radiation therapy is not feasible after adjuvant AC chemotherapy for early-stage breast cancer. Concurrent treatment using a less frequent paclitaxel dosing schedule may be possible, but caution is warranted in light of the apparent possibility of pulmonary injury. PMID- 16243444 TI - Determining optimal clinical target volume margins in head-and-neck cancer based on microscopic extracapsular extension of metastatic neck nodes. AB - PURPOSE: To determine the optimal clinical target volume margins around the gross nodal tumor volume in head-and-neck cancer by assessing microscopic tumor extension beyond cervical lymph node capsules. METHODS AND MATERIALS: Histologic sections of 96 dissected cervical lymph nodes with extracapsular extension (ECE) from 48 patients with head-and-neck squamous cell carcinoma were examined. The maximum linear distance from the external capsule border to the farthest extent of the tumor or tumoral reaction was measured. The trends of ECE as a function of the distance from the capsule and lymph node size were analyzed. RESULTS: The median diameter of all lymph nodes was 11.0 mm (range: 3.0-30.0 mm). The mean and median ECE extent was 2.2 mm and 1.6 mm, respectively (range: 0.4-9.0 mm). The ECE was <5 mm from the capsule in 96% of the nodes. As the distance from the capsule increased, the probability of tumor extension declined. No significant difference between the extent of ECE and lymph node size was observed. CONCLUSION: For N1 nodes that are at high risk for ECE but not grossly infiltrating musculature, 1 cm clinical target volume margins around the nodal gross tumor volume are recommended to cover microscopic nodal extension in head and-neck cancer. PMID- 16243445 TI - Intraoperative electron-beam radiotherapy and ureteral obstruction. AB - PURPOSE: To quantify the risk of ureteral obstruction (UO) after intraoperative electron-beam radiotherapy (IOERT). METHODS AND MATERIALS: One hundred forty-six patients received IOERT of 7.5 to 30 Gy to 168 ureters; 132 patients received external radiotherapy. RESULTS: Follow-up ranged from 0.01 to 19.1 years (median, 2.1 years). The rates of clinically apparent type 1 UO (UO from any cause) after IOERT at 2, 5, and 10 years were 47%, 63%, and 79%, respectively. The rates of clinically apparent type 2 UO (UO occurring at least 1 month after IOERT, excluding UO caused by tumor or abscess and patients with stents) at 2, 5, and 10 years were 27%, 47%, and 70%, respectively. Multivariate analysis revealed that the presence of UO before IOERT (p < 0.001) was associated with an increased risk of clinically apparent type 1 UO. Increasing IOERT dose (p < 0.04) was associated with an increased risk of clinically apparent type 2 UO. UO rates in ureters not receiving IOERT at 2, 5, and 10 years were 19%, 19%, and 51%, respectively. CONCLUSIONS: Risk of UO after IOERT increases with dose. However, UO risk for ureters not receiving IOERT was also high, which suggests an underlying risk of ureteral injury from other causes. PMID- 16243446 TI - Long-term outcome and mortality trends in early-stage, Grade 1-2 follicular lymphoma treated with radiation therapy. AB - PURPOSE: To analyze long-term outcomes and causes of death in patients receiving radiation therapy (RT) for localized, low-grade follicular lymphoma. METHODS AND MATERIALS: Between 1972 and 2000, 106 patients with Stage I-II, Grade 1-2 follicular lymphoma received RT alone or radiation and chemotherapy (RT/CT). Seventy-four percent had Stage I, and 26% had Stage II disease. Seventy-six percent received RT alone, and 24% received combined RT/CT. Second malignancy rates were compared with an age- and sex-matched population. RESULTS: Median follow-up was 12 years. Median survival time was 19 years. The 5-, 10-, and 15 year overall survival (OS) rates were 93%, 75%, and 62%, respectively. Age > or = 60 was the only significant adverse prognostic factor with respect to OS. There were 35 deaths, 20 of which were attributable to lymphoma. Freedom from treatment failure (FFTF) rates at 5, 10, and 15 years were 72%, 46%, and 39%, respectively. Forty-seven patients (48%) relapsed. Tumor size > 3 cm was the only significant adverse factor for FFTF. Observed incidence of second malignancy did not significantly exceed expected incidence. CONCLUSIONS: Although patients with early-stage, low-grade follicular lymphoma have long median survival, the leading cause of death remains lymphoma. However, patients receiving RT do not have significantly elevated cumulative incidence of second malignancy. PMID- 16243447 TI - Antifungal effect of some spice hydrosols. AB - The antifungal effects of rosemary, cumin, sater (savory), basil and pickling herb hydrosols were investigated against Rhizoctonia solani, Fusarium oxysporum f. sp tulipae, Botrytis cinerea and Alternaria citri. Hydrosols of sater and pickling herb showed the most relevant fungicidal activity. PMID- 16243448 TI - Protective effect of Piper longum fruit ethanolic extract on radiation induced damages in mice: a preliminary study. AB - The radioprotective property of an ethanolic extract of Piper longum fruits (EEPLF) was investigated in Swiss mice. The white blood cell (WBC) count in irradiated control mice was drastically reduced to 1900 cells/mm3 on third day but in treated animals the count was 2783.3 cells/mm3. The number of bone marrow cells and alpha-esterase positive cells was also enhanced by the EEPLF administration (16.7 x 10(6) cells/femur and 946.5/4000 cells, respectively) when compared to the radiation exposed control animals (12.2 x 10(6) cells/femur and 693.5/4000 cells, respectively). EEPLF reduced the elevated levels of glutathione pyruvate transaminase (GPT), alkaline phosphatase (ALP), and lipid peroxidation (LPO) in liver and serum of radiation treated animals. The extract administration also increased the reduced glutathione (GSH) production to offer the radioprotection. PMID- 16243449 TI - Participation of the recA determinant in the transposition of class II transposon mini-TnMERI1. AB - As an initial step to understand the mobile nature of class II mercury resistance transposon TnMERI1, the effect of the recA gene on translocation of mini-TnMERI1 was evaluated. A higher transposition frequency in the LE392 strain (2.4+/ 1.2x10(-5)) than in the recA-deficient DH1 strain (1.2+/-0.8x10(-6)) indicated participation of the recA gene in mini-TnMERI1 transposition. Introduction of the recA gene into the DH1 strain complemented the transposition frequency at the same level as in LE392 and confirmed participation of the recA gene in transposition. However, treatment of cells by stress agents, including irradiation of up to 3000 Jm(-2) UV doses, did not alter the transposition frequency and suggested independence of RecA from the SOS stress response. Further analysis of transconjugants indicated participation of RecA in the resolution of the cointegrate structure of the transposon. These results suggested that RecA is a constitutive cellular factor that increases translocation of mini-TnMERI1 and may participate in dissemination of TnMERI1 like transposons. PMID- 16243450 TI - Use of a microarray to assess the distribution of plasmid and chromosomal virulence genes in strains of enteroaggregative Escherichia coli. AB - A DNA microarray was used to analyze the distribution of plasmid and chromosomal genes among strains of enteroaggregative Escherichia coli (EAEC) isolated from a prospective diarrhoea surveillance study in the United Kingdom. Target genes were extracted from existing databases and from the genome sequence of prototype EAEC strain 042. We found that strains exhibiting the aggregative adherence (AA) phenotype could be broadly divided into two groups depending upon whether they harboured genes from the EAEC virulence plasmid (pAA) and a set of chromosomal genes found in EAEC strain 042. Several chromosomal loci were inherited en bloc, and were more common in strains which we designated Group 1; genes at the pheU locus were particularly conserved. Genes encoded on the pAA plasmid and those under control of the master regulator AggR were also concentrated in the Group 1 EAEC. A gene encoding a type 1 pilin allele was detected more frequently in Group 2 EAEC. Our data suggest that strains previously designated as typical EAEC harbour a large number of conserved plasmid and chromosomal loci, further illuminating a package of virulence genes common to the most important EAEC. PMID- 16243451 TI - Agrobacterium tumefaciens-mediated transformation of Leptosphaeria spp. and Oculimacula spp. with the reef coral gene DsRed and the jellyfish gene gfp. AB - Four filamentous ascomycetes, Leptosphaeria maculans, L. biglobosa, Oculimacula yallundae and O. acuformis, were transformed via Agrobacterium tumefaciens mediated transformation with the genes encoding DsRed and GFP. Using vectors pCAMDsRed and pCAMBgfp, either germinated conidia of Leptosphaeria spp. and O. yallundae or physically fragmented cultures of Oculimacula spp. were transformed. In vitro, the expression of the two reporter proteins in mycelium of both Oculimacula and both Leptosphaeria species was sufficient to distinguish each species in co-inoculated cultures. In planta, transformants of L. maculans or L. biglobosa expressing DsRed or GFP could be observed together in leaves of Brassica napus. Either reporter protein could be used to view the colonization of leaf petioles by both Leptosphaeria spp. and growth in the xylem vessels could be clearly observed. With the generation of these transformants, further studies on interactions between pathogen species involved in disease complexes on various host species and between opposite mating types of the same species are now possible. PMID- 16243452 TI - Reversible membrane association of dinitrogenase reductase activating glycohydrolase in the regulation of nitrogenase activity in Rhodospirillum rubrum; dependence on GlnJ and AmtB1. AB - In the photosynthetic bacterium Rhodospirillum rubrum nitrogenase activity is regulated by reversible ADP-ribosylation of dinitrogenase reductase in response to external so called "switch-off" effectors. Activation of the modified, inactive form is catalyzed by dinitrogenase reductase activating glycohydrolase (DRAG) which removes the ADP-ribose moiety. This study addresses the signal transduction between external effectors and DRAG. R. rubrum, wild-type and P(II) mutant strains, were studied with respect to DRAG localization. We conclude that GlnJ clearly has an effect on the association of DRAG to the membrane in agreement with the effect on regulation of nitrogenase activity. Furthermore, we have generated a R. rubrum mutant lacking the putative ammonium transporter AmtB1 which was shown not to respond to "switch-off" effectors; no loss of nitrogenase activity and no ADP-ribosylation. Interestingly, DRAG was mainly localized to the cytosol in this mutant. Overall the results support our model in which association to the membrane is part of the mechanism regulating DRAG activity. PMID- 16243453 TI - The Trichoderma reesei hydrophobin genes hfb1 and hfb2 have diverse functions in fungal development. AB - Hydrophobins are fungal self-assembling proteins. Here, the hydrophobin genes hfb1 and hfb2 were deleted in Trichoderma reesei and their biological roles studied. Our results suggest that HFBI has a role in hyphal development and HFBII in sporulation. Sporulating colonies of the Deltahfb2 strain were wettable and sporulation was only 50% of the parent strain. Colonies of Deltahfb1 showed wettable and fluffy phenotype. In shaken liquid cultures, the hyphae of Deltahfb1 were thinner and biomass formation was slower compared to the parent strain while in static liquid cultures no aerial hyphae were formed. Expressing the Schizophyllum commune hydrophobin SC3 in the Deltahfb1 strain restored the formation of aerial hyphae. PMID- 16243454 TI - Isolation of a triazophos-degrading strain Klebsiella sp. E6 effectively utilizing triazophos as sole nitrogen source. AB - A triazophos-degrading strain, Klebsiella sp. E6, was isolated by enrichment technology from soil that had been exposed long-term to triazophos. The strain grew well at pH 7.0-8.0 with a broad temperature profile ranging from 32 to 37 degrees C. It could keep good growth on methanol as carbon source and TAP as additional carbon source or nitrogen source. The experiment on the degradation activities of strain E6 showed that it utilized TAP more effectively when TAP was supplied as the sole nitrogen source, as opposed to additional carbon source. The intermediates of triazophos metabolism indicated that degradation occurred through a hydrolysis mechanism, one of the products of which, 1-phenyl-3-hydroxy 1,2,4-triazole, was also mineralized by strain E6. PMID- 16243455 TI - Purification and properties of a xylanase from Ceriporiopsis subvermispora cultivated on Pinus taeda. AB - The production of hemicellulose and cellulose degrading enzymes by the white-rot fungus Ceriporiopsis subvermispora was determined while growing in Pinus taeda wood chips. Enzymes produced by the fungus were extracted after 30 days of cultivation and at least two different xylanases were secreted. An endo-(1,4) beta-xylanase was purified by means of ultrafiltration, anion exchange chromatography and gel filtration. Its molecular mass was 29 kDa and the pH and temperature optima were 5.0 and 60 degrees C, respectively. The endo-xylanase was able to hydrolyze xylan to principally xylotriose and xylotetraose and it has different activities against different xylans. With birchwood xylan as substrate, the enzyme showed a K(m) of 1.93 mg/ml and specific activity of 538 units/mg protein at 50 degrees C. PMID- 16243456 TI - Characteristics of Streptococcus mutans strains lacking the MazEF and RelBE toxin antitoxin modules. AB - Two pairs of genes were identified in Streptococcus mutans with similarity to relBE and mazEF toxin-antitoxin (TA) modules of Escherichia coli. Transcription of mazEF and relBE was repressed by amino acid starvation, and relBE expression was repressed by low pH. Mutants lacking MazF, RelE, or both toxins (MRT1) grew in broth media and formed biofilms as well as the parent. Biofilm populations of MRT1 were more resistant to acid killing than the parent or single mutants. MRT1 also exhibited a longer diauxie during growth on glucose and inulin and displayed decreased phosphoenolpyruvate:sugar phosphotransferase activity. This is the first report that demonstrates a physiological role for TA modules in Gram positive bacteria. PMID- 16243457 TI - Homologous recombination of exogenous DNA with the Sulfolobus acidocaldarius genome: properties and uses. AB - In order to quantify recombination between exogenous DNA and the Sulfolobus acidocaldarius chromosome, we electroporated pyrE (uracil-auxtotrophic) recipient strains with functional pyrE sequences and counted Pyr+ transformants by direct plating. Certain culture and post-electroporation conditions increased the yield of Pyr+ recombinants from non-replicating pyrE plasmid, whereas cognate methylation of SuaI restriction sites in the plasmid decreased it. Recombination of linear DNAs with the S. acidocaldarius genome was proportional to the length of a limiting overlap, but even synthetic oligonucleotides produced reasonable numbers of recombinants with appropriate recipient strains. To investigate uses of this latter property, we electroporated an 18-bp pyrE deletion mutant with mixtures of synthetic oligonucleotides altering glycine-55 of the orotate phosphoribosyl transferase encoded by pyrE. Pyr+ transformants were recovered in which this codon was converted to each of the alternatives encoded by the oligonucleotide mixtures, thereby identifying five amino acid substitutions tolerated at this position of the thermostable enzyme. PMID- 16243458 TI - Antimalarial drug synergism and antagonism: mechanistic and clinical significance. AB - Interactions between antimicrobial agents provide clues as to their mechanisms of action and influence the combinations chosen for therapy of infectious diseases. In the treatment of malaria, combinations of drugs, in many cases acting synergistically, are increasingly important in view of the frequency of resistance to single agents. The study of antimalarial drug interactions is therefore of great significance to both treatment and research. It is therefore worrying that the analysis of drug-interaction data is often inadequate, leading in some cases to dubious conclusions about synergism or antagonism. Furthermore, making mechanistic deductions from drug-interaction data is not straightforward and of the many reported instances of antimalarial synergism or antagonism, few have been fully explained biochemically. This review discusses recent findings on antimalarial drug interactions and some pitfalls in their analysis and interpretation. The conclusions are likely to have relevance to other antimicrobial agents. PMID- 16243459 TI - Does sympathetic activity contribute to growth of preterm infants? AB - BACKGROUND: Emerging evidence on faster growth in preterm infants with higher heart rate as opposed to the classical approach calls for further research. AIMS: to test whether (1) high or low heart rate and (2) heart rate during the first days of life predict greater weight gain in preterm infants. METHODS: A retrospective study analyzing two daily measures of heart rate obtained during restful sleep, total daily calorie intake and daily weight gain measured always in the morning before meal were collected from the medical files. RESULTS: Analysis of 90 healthy preterm infants born at 32-36 weeks of gestation revealed that increased mean heart rate during hospitalization predicted greater weight gain even when controlling for calorie intake, birth weight, gestational age, appropriateness of birth weight for gestational age, and length of hospitalization. Mean heart rate during the first three days of life yielded the same pattern of results. Post-hoc analysis of variance between infants with mean daily heart rate /=140 bpm showed that infants with higher heart rate achieved a significantly higher weight gain. CONCLUSIONS: It is suggested that, contrary to adults, in neonates an anabolic activity is represented by increased sympathetic functioning within the normal range. The implications of a slower growth rate for additional developmental care and individual considerations of appropriate stimulation in preterm infants are discussed. PMID- 16243460 TI - Repeated topical treatment, in contrast to single oral doses, with Vitamin A containing preparations does not affect plasma concentrations of retinol, retinyl esters or retinoic acids in female subjects of child-bearing age. AB - BACKGROUND: Vitamin A is widely used in cosmetic preparations. Given that oral Vitamin A and its metabolites present a potential reproductive risk, the present study investigated the effect of topical Vitamin A on human endogenous plasma levels of Vitamin A and its metabolites. METHODS: Two groups of 14 female volunteers of child-bearing age were kept on a Vitamin A-poor diet and treated topically for 21 days with creams containing 0.30% retinol or 0.55% retinyl palmitate on approximately 3000 cm2 of their body surface area, amounting to a total of approximately 30,000 IU Vitamin A/subject/day. After a 12-day wash-out period, the study groups received single oral doses of 10,000 IU or 30,000 IU retinyl palmitate (RP), corresponding to the maximal EU allowance during pregnancy or three-times higher, respectively. Blood samples were collected over 24h on study days -3 (pre-study), 1, 21 (first and last days of topical treatment) and 34 (oral administration) at 0, 1, 2, 4, 6, 8, 12, 14-16 h and 24 h after treatment for determination of plasma concentrations of retinol (REL), retinyl palmitate (RP), oleate (RO) and stearate (RS), 9-cis-, 13-cis-, all-trans (AT), 13-cis-4-oxo- or AT-4-oxo-retinoic acids (RAs). RESULTS: With the exception of transient mild (RP-group) to moderate (REL-group) local irritation on the treatment sites, no adverse local or systemic effects were noted. On days 1 or 21 of topical treatment, no changes were measured in individual or group mean plasma Cmax, AUC0-24 h or other pharmacokinetic parameters of REL, retinyl esters or RAs relative to pre-study data. In contrast, single oral doses of RP at 10,000 IU or 30,000 IU produced dose-related and sustained increases in Cmax and AUC0-24 h values of plasma RP, RO, RS, 13-cis- and 13-cis-4-oxo-RAs, as well as a transient increase in AT-RA. In conclusion, our results provide evidence that human topical exposure to retinol- or retinyl ester-containing cosmetic creams at 30,000 IU/day and maximal use concentrations do not affect plasma levels of retinol, retinyl esters or RAs, whereas single oral doses at 10,000 IU or 30,000 IU produce significant increases in plasma retinyl esters and RAs. PMID- 16243461 TI - Autosomal recessive and sporadic deafness in Morocco: high frequency of the 35delG GJB2 mutation and absence of the 342-kb GJB6 variant. AB - Deafness is a heterogeneous disorder showing different pattern of inheritance and involving a multitude of different genes. Mutations in the gene, GJB2 Gap junction type 1), encoding the gap junction protein connexin-26 on chromosome 13q11 may be responsible for up 50% of autosomal recessive nonsyndromic hearing loss cases (ARNSHL), and for 15-30% of sporadic cases. However, a large proportion (10-42%) of patients with GJB2 has only one GJB2 mutant allele. Recent reports have suggested that a 342-kb deletion truncating the GJB6 gene (encoding connexin-30), was associated with ARNSHL through either homozygous deletion of Cx30, or digenic inheritance of a Cx30 deletion and a Cx26 mutation in trans. Because mutations in Connexin-26 (Cx26) play an important role in ARNSHL and that distribution pattern of GJB2 variants differs considerably among ethnic groups, our objective was to find out the significance of Cx26 mutations in Moroccan families who had hereditary and sporadic deafness. One hundred and sixteen families with congenital deafness (including 38 multiplex families, and 78 families with sporadic cases) were included. Results show that the prevalence of the 35delG mutation is 31.58% in the family cases and 20.51% in the sporadic cases. Further screening for other GJB2 variants demonstrated the absence of other mutations; none of these families had mutations in exon 1 of GJB2 or the 342-kb deletion of GJB6. Thus, screening of the 35delG in the GJB2 gene should facilitate routinely used diagnostic for genetic counselling in Morocco. PMID- 16243462 TI - Expression of bFGF and NGF and their receptors in chick's auditory organ following overexposure to noise. AB - Growth factors are known to activate signaling cascades for DNA replication; they participate in the regulation of cell differentiation and are required as positive signals for cell survival. Thus, many of them may be regarded as potential candidates stimulating regeneration processes in the inner ear. We analyzed the expression of basic fibroblast growth factor (bFGF) and nerve growth factor (NGF) and their receptor (bFGFR and NGFR)-like immunoreactivity in chick basilar papillae, along with bFGF and NGF mRNA expression. The evaluation was made 1 and 5 days after exposure to wide-band noise with two increasing levels of acoustic energy. For both factors, the immunoreactivity was shown predominantly in the middle part of basilar papilla, in noise-exposed, but not control birds. It was localized in the cytoplasm of hair cells, nuclei of supporting cells and cytoplasm of ganglion cells. Strong immunoreactivity of bFGFR and NGFR was found both in control and noise-exposed animals, with the cell localization similar to that of growth factors. The increase in mRNA expression for bFGF and NGF was found in noise-exposed animals only after lower exposure to noise, on day 5 after exposure (p<0.01). A lack of increased expression after higher exposure could be excused by larger damage of hair cells followed by the increase of mRNA for beta actin to which the results were referred. The results suggest bFGF and NGF involvement in postinjury regeneration of the basilar papilla. PMID- 16243463 TI - Phytochemical screening and effect of aqueous extract of Ficus sycomorus L. (Moraceae) stembark on muscular activity in laboratory animals. AB - The stembark of Ficus sycomorus was collected, dried and extracted, to screen for some chemical constituents and study its effect on muscle contraction. The duodena and recti abdominis of 10 guinea pigs weighing between 330 and 345 g and 10 frogs weighing between 180 and 201 g, respectively, were isolated and used for this study. The extract was tested to see its effect on acetylcholine (ACH) induced contraction on kymograph. The extract reduced the acetylcholine contractile responses of guinea pigs duodena and recti abdominis muscles of frogs significantly, thus showing inhibitory effect on muscle contraction. The extract showed the presence of gallic tannins, saponins, reducing sugars, alkaloids and flavone aglycones. It was concluded that the extract has inhibitory effect on both smooth and skeletal muscles contractions and contains important constituents for pharmacological activities. PMID- 16243464 TI - Nitric oxide-dependent vasorelaxation induced by extractive solutions and fractions of Maytenus ilicifolia Mart ex Reissek (Celastraceae) leaves. AB - This study reveals that an ethanolic supernatant obtained from an aqueous extractive solution prepared from residues of methanolic extracts of ground leaves of Maytenus ilicifolia is able to cause a concentration- and endothelium dependent relaxation in pre-contract rat aorta rings, with EC(50) of 199.7 (190 210) microg/ml. The non-selective nitric oxide synthase inhibitors l-NAME and l NMMA abolished this effect, while superoxide dismutase and MnTBAP (a non enzymatic superoxide dismutase mimetic) enhanced it. Further, relaxation induced by this ethanolic supernatant have been strongly inhibited by the guanylate cyclase inhibitors methylene blue and ODQ, as well as by the potassium channel blockers 4-aminopyridine and tetraethylammonium, but was unchanged by the cyclooxygenase inhibitor indomethacin and the membrane receptor antagonists atropine, HOE-140 and pirilamine. Partition of the ethanolic supernatant between H(2)O and EtOAc generated a fraction several times more potent, able to fully relax endothelium-intact aorta rings with an EC(50) of 4.3 (3.9-4.8) microg/ml. (13)C NMR spectrum of this fraction showed signals typical of catechin. This study reveals that the leaves of M. ilicifolia possess one or more potent substances able to relax endothelium-intact rat aorta rings, an event that appears to involve nitric oxide production, guanylate cyclase activation and potassium channel opening. PMID- 16243465 TI - Chemical composition and biological activities of essential oil from the leaves of Sesuvium portulacastrum. AB - Sesuvium portulacastrum has long been used as a remedy for fever and scurvy. Hydrodistillation was used to extract the essential oil from the fresh leaves of Sesuvium portulacastrum. The essential oil yield obtained was 0.15%. Using GC-MS analysis, alpha-pinene, camphene, beta-pinene, alpha-terpinene, O-cymene, limonene, 1,8-cineole, alpha-terpinene, bornyl acetate, tridecane, trans caryophyllene and alpha-humulene were identified. The hole plate diffusion method was used for antibacterial testing. The essential oil exhibited antibacterial activity against Acetobacter calcoacetica, Bacillus subtillis, Clostridium sporogenes, Clostridium perfringens, Escherichia coli, Salmonella typhii, Staphylococcus aureus and Yersinia enterocolitica. The mycelium growth inhibition method was used for the antifungal testing. The oil exhibited antifungal activity against Candida albicans, Aspergillus niger, Aspergillus flavus and Penicillium notatum. Using the beta-carotene, acetone and linoleic acid method for the antioxidant testing, the essential oil showed antioxidant activity threshold of 15.9 mm mean zone of color retention. PMID- 16243466 TI - Opuntia ficus-indica attenuates neuronal injury in in vitro and in vivo models of cerebral ischemia. AB - We examined whether the methanol extract of Opuntia ficus-indica (MEOF) has a neuroprotective action against N-methyl-d-aspartate (NMDA)-, kainate (KA)-, and oxygen-glucose deprivation (OGD)-induced neuronal injury in cultured mouse cortical cells. We also evaluated the protective effect of MEOF in the hippocampal CA1 region against neuronal damage evoked by global ischemia in gerbils. Treatment of neuronal cultures with MEOF (30, 300, and 1000 microg/ml) inhibited NMDA (25 microM)-, KA (30 microM)-, and OGD (50 min)-induced neurotoxicity dose-dependently. The butanol fraction of Opuntia ficus-indica (300 microg/ml) significantly reduced NMDA (20 microM)-induced delayed neurotoxicity by 27%. Gerbils were treated with MEOF every 24h for 3 days (0.1, 1.0, and 4.0 g/kg, p.o.) or for 4 weeks (0.1 and 1.0 g/kg, p.o.), and ischemic injury was induced after the last dose. Neuronal cell damage in the hippocampal CA1 region was evaluated quantitatively at 5 days after the ischemic injury. When gerbils were given doses of 4.0 g/kg (3 days) and 1.0 g/kg (4 weeks), the neuronal damage in the hippocampal region was reduced by 32 and 36%, respectively. These results suggest that the preventive administration of Opuntia ficus-indica extracts may be helpful in alleviating the excitotoxic neuronal damage induced by global ischemia. PMID- 16243468 TI - Soleus and EDL muscle contractility across the lifespan of female C57BL/6 mice. AB - All previous aging research on the contractility of rodent skeletal muscle has been conducted on male rodents. Because males and females age differently, we undertook this study to determine if and when age-related decrements in skeletal muscle contractility occur in female mice. Soleus and extensor digitorum longus (EDL) muscles from female C57BL/6 mice aged approximately 4, 8, 16, 24 and 28 mo were assessed in vitro for contractility and subsequently contractile protein content. EDL muscle was resistant to age-related changes in force generation but displayed characteristics of becoming more slow-twitch like. Maximal isometric tetanic force (Po) generated by soleus muscle declined with age. Soleus muscle size and contractile protein contents were not affected by age and thus could not explain the age-related force decrements. Soleus muscle specific Po declined with age being approximately 26% lower in muscles of 16-28 mo-old mice indicating that a deterioration in soleus muscle quality of female mice occurred beginning around the age of ovarian failure. Thus this study provides essential, comprehensive baseline data for future studies on age-related muscle dysfunction in the female mouse. PMID- 16243467 TI - Results of the 2003-2004 GEP-ISFG collaborative study on mitochondrial DNA: focus on the mtDNA profile of a mixed semen-saliva stain. AB - We report here a review of the seventh mitochondrial DNA (mtDNA) exercise undertaken by the Spanish and Portuguese working group (GEP) of the International Society for Forensic Genetics (ISFG) corresponding to the period 2003-2004. Five reference bloodstains from five donors (M1-M5), a mixed stain of saliva and semen (M6), and a hair sample (M7) were submitted to each participating laboratory for nuclear DNA (nDNA; autosomal STR and Y-STR) and mtDNA analysis. Laboratories were asked to investigate the contributors of samples M6 and M7 among the reference donors (M1-M5). A total of 34 laboratories reported total or partial mtDNA sequence data from both, the reference bloodstains (M1-M5) and the hair sample (M7) concluding a match between mtDNA profiles of M5 and M7. Autosomal STR and Y STR profiling was the preferred strategy to investigate the contributors of the semen/saliva mixture (M6). Nuclear DNA profiles were consistent with a mixture of saliva from the donor (female) of M4 and semen from donor M5, being the semen (XY) profile the dominant component of the mixture. Strikingly, and in contradiction to the nuclear DNA analysis, mtDNA sequencing results yield a more simple result: only the saliva contribution (M4) was detected, either after preferential lysis or after complete DNA digestion. Some labs provided with several explanations for this finding and carried out additional experiments to explain this apparent contradictory result. The results pointed to the existence of different relative amounts of nuclear and mtDNAs in saliva and semen. We conclude that this circumstance could strongly influence the interpretation of the mtDNA evidence in unbalanced mixtures and in consequence lead to false exclusions. During the GEP-ISFG annual conference a validation study was planned to progress in the interpretation of mtDNA from different mixtures. PMID- 16243469 TI - Quantitative assay of lorazepam and its metabolite glucuronide by reverse-phase liquid chromatography-tandem mass spectrometry in human plasma and urine samples. AB - A LC/MS/MS method for the quantitative determination of lorazepam in human plasma and urine samples was developed and validated. The enantioselective assay allowed to separate the enantiomers and to verify the stereochemical instability of lorazepam. The linearity assessed for lorazepam unchanged was 0.2-20 ng of each enantiomer/ml plasma and 0.2-15 ng of each enantiomer/ml urine. The linearity assessed for total lorazepam (after enzymatic hydrolysis) was 1-30 ng of each enantiomer/ml plasma and 10-150 ng of each enantiomer/ml urine. The coefficients of variation obtained for the intra- and interassay precision were less than 15%. The method was applied to the investigation of the kinetic disposition and metabolism of racemic lorazepam administered as a single oral dose of 2 mg to a parturient. The occurrence of racemization required the calculation of the pharmacokinetic parameters as enantiomeric mixtures of lorazepam (t(1/2a) 3.5h; K(a) 0.198 ngh(-1); t(1/2) 11.5h; beta 0.060 h(-1); AUC(0-infinity) 192.1ngh/ml; CLt/f 2.41ml/minkg; Vd/f 173.5l; Fel 0.41%, and Cl(R) 0.0099 ml/minkg) and its metabolite lorazepam-glucuronide (t(1/2f) 1.2h; K(f) 0.578 h(-1); t(1/2) 16.6h; beta 0.042 h(-1); AUC(0-infinity) 207.6 ngh/ml; Fel 51.80%, and Cl(R) 98.32 ml/minkg). However, the determined confidence limits make the method suitable for application to clinical pharmacokinetic studies, even if the quantification of both the enantiomers is required. PMID- 16243470 TI - Determination of tobramycin and impurities using high-performance anion exchange chromatography with integrated pulsed amperometric detection. AB - Tobramycin is one of a class of aminoglycoside antibiotics that lack a good chromophore, and is therefore difficult to determine using reversed-phase HPLC with absorbance detection. This is especially true for determining the quantity of each impurity. We show that tobramycin and its major impurities, including kanamycin B and neamine (neomycin A), can be separated on a strong anion-exchange column using a weak potassium hydroxide eluent (2.00 mM) at a column temperature of 30 degrees C, and directly detected by integrated pulsed amperometric detection (IPAD). The resolution (United States Pharmacopeia (USP) definition) between tobramycin and kanamycin B ranged from 5.71 and 6.06 over 7 days of consecutive analysis (5.92+/-0.07, n = 590 injections). Due to the difficulty of producing weak hydroxide eluents of the required purity (i.e. carbonate-free), this method depends on automatic eluent generation to ensure method ruggedness. This method exhibited good long-term (50 days, 2368 injections) retention time stability with R.S.D.s of 0.4% and 0.3% for tobramycin and kanamycin B, respectively. Peak area R.S.D.s for tobramycin and kanamycin B (10 microM each, 20 microL injection) over 7 days (572 injections) were 2.3% and 1.9%, respectively. Method robustness was evaluated by intentionally varying the flow rate, eluent concentration, column temperature, and column. Based on the results of these evaluations, this method can be used for tobramycin identity, assay, and purity. PMID- 16243471 TI - Determination of albuterol sulfate and its related substances in albuterol sulfate inhalation solution, 0.5% by RP-HPLC. AB - An isocratic reversed-phase high performance liquid chromatographic (RP-HPLC) method has been developed and validated for the determination of albuterol sulfate and six of its related substances in albuterol sulfate inhalation solution, 0.5% (w/v). The separation was achieved using a YMC phenyl column (250 mm x 4.6 mm ID, 5 microm fitted with a direct connect YMC phenyl guard column (20 mm x 4 mm ID) maintained at ambient conditions, and a mobile phase of 25 mM monobasic potassium phosphate (pH 3.0) and methanol (95:5, v/v). The mobile phase flow rate was 1.5 mL/min and the detection wavelength was 225 nm. Albuterol is quantitated versus an external standard. The method was capable of resolving six of the seven known albuterol-related substances. Bis-ether albuterol, a drug substance process related impurity, is retained on the column due to its different hydrophilic character. The related substances are determined by area percent. However, a correction factor of 1.6 is applied for the determination of albuterol aldehyde, a potential impurity and a degradation product, since its molar absorptivity is about 1.6 times that of albuterol. The limits of detection and quantitation for albuterol and six of its related substances ranged between 0.01 and 0.21% of the assay concentration of 0.3 mg/mL as albuterol base. The method was found to be linear for albuterol over the range of 50-150% of the active label claim. The method was also found to be linear for the six related substances over the range 0.05-0.5%. No interferences from the blank, placebo (formulation matrix), related substances or force-degraded placebo samples were observed for the determination of the active or the individual related substances. The method was found to be accurate, precise, linear, specific, sensitive, rugged, robust, and stability-indicating. PMID- 16243473 TI - Rothia dentocariosa endocarditis complicated by multiple cerebellar hemorrhages. AB - A rare case of Rothia dentocariosa endocarditis after oral surgery is presented. The patient had repeated cerebellar hemorrhages occurring over months as the sole manifestation of disease. He was successfully treated with penicillin G and gentamicin with good outcome. PMID- 16243472 TI - Multilocus sequence typing and repetitive element-based polymerase chain reaction analysis of Neisseria meningitidis isolates in Brazil reveal the emergence of 11 new sequence types genetically related to the ST-32 and ST-41/44 complexes and high prevalence of strains related to hypervirulent lineages. AB - Eighty-four strains of Neisseria meningitidis isolated from patients with meningococcal disease in 4 states of Brazil were analyzed by multilocus sequence typing and repetitive element-based polymerase chain reaction (Rep-PCR). The majority of strains analyzed (82%) belonged to 4 hypervirulent lineages, and 11 of 20 new sequence type (STs) characterized were related to hypervirulent lineages. Sequences of fetA and porA genes were analyzed, and the majority were related to profiles present in the ST-32 complex/electrophoretic type (ET)-5 complex. Rep-PCR analysis showed a unique electrophoretic pattern among strains related to hypervirulent lineages. Considering that 81% of the strains were serogroup B and strains belonging to the ST-32/ET-5 complex are genetically related to the Cuban vaccine strain used in a mass vaccination from 1990 to 1994 in Brazil, we believe that this vaccine did not confer effective herd immunity even among the age group within the vaccine showed higher efficacy. Our results once more raise the question about which strains should be used in the development of a new vaccine against N. meningitidis serogroup B. PMID- 16243474 TI - Evaluation of dipicolinic acid for detection of IMP- or VIM- type metallo-beta lactamase-producing Pseudomonas aeruginosa clinical isolates. AB - We evaluated dipicolinic acid (DPA) as a chelating agent for detection of IMP- or VIM-type metallo-beta-lactamase (MBL)-producing Pseudomonas aeruginosa clinical isolates. Using the broth microdilution testing in the presence or absence of DPA, MBL producers exhibited 100%, 92%, or 100% of >or=8 times (media, 32 times) reduction of MICs in presence of DPA for ceftazidime, imipenem, or meropenem, respectively. In disk diffusion testing, expansion of growth inhibitory zone of these clinical isolates was clearly observed. Thus, DPA could be useful in the detection for MBL-producing P. aeruginosa clinical isolates. PMID- 16243475 TI - Development of a whole-virus multiplex flow cytometric assay for antibody screening of a specific pathogen-free primate colony. AB - Our goal was to determine if a multiplex technique using a fluorescent bead-based flow cytometric assay could yield results comparable to traditional enzyme-linked immunosorbent assay (ELISA) in terms of sensitivity, specificity, cross reactivity, and throughput. We applied both techniques to serologic screening of specific pathogen-free macaques, for type D simian retrovirus, simian T lymphotropic virus, Cercopithicine herpesvirus 1, and simian immunodeficiency virus, and found a high correlation between the bead-based multiplex assay and ELISA. The multiplex assay demonstrated greater sensitivity with no loss in specificity when compared to the ELISA. A lower false-positive rate with the multiplex assay decreased the number of confirmatory Western blots required. Using the multiplex assay, we were able to screen samples for 4 viruses simultaneously in the time it took to perform a single-virus ELISA, resulting in a faster turnaround time and higher throughput. The multiplexed assay provided greater sensitivity, increased stability, and better performance than ELISA. PMID- 16243476 TI - Changing patterns of fluoroquinolone resistance among Bacteroides fragilis group organisms over a 6-year period (1997-2002). AB - The evolution of susceptibility to the newer fluoroquinolones, moxifloxacin and trovafloxacin, of Bacteroides fragilis group organisms isolated in our hospital from 1997 to 2002 was studied. A total of 927 strains were included in the study. Trovafloxacin was more active than moxifloxacin against the various species of the group. During the study period, an increase in resistance to both quinolones was observed. Rates of resistance to moxifloxacin at a breakpoint of 8 microg/mL remained stable at around 6% during the period 1997-1998 and increased to 11.4% in 2000 and to 16.5% in 2001-2002 (P<0.005). Resistance to trovafloxacin rose significantly from 0.6% in 1998 to 6.8% in 1999 (P<0.05) and did not change appreciably over the last 3 years studied. This study confirms the increasing resistance of B. fragilis group organisms to trovafloxacin and moxifloxacin and emphasizes the need to perform periodic antimicrobial susceptibility tests to guide the selection of appropriate antimicrobial therapy. PMID- 16243477 TI - Simultaneous detection of isoniazid, rifampin, and ethambutol resistance of Mycobacterium tuberculosis by a single multiplex allele-specific polymerase chain reaction (PCR) assay. AB - Prompt detection of drug resistance of Mycobacterium tuberculosis is essential for effective control of tuberculosis (TB). We developed a multiplex allele specific polymerase chain reaction (MAS-PCR) that detects the most commonly observed isoniazid (INH), rifampin (RIF), and ethambutol resistance-associated mutations in a single assay. The usefulness of the newly developed method was evaluated with 174 clinical isolates of M. tuberculosis obtained from Turkey. Distinct PCR banding patterns were observed for different mutation profiles and the correlation between MAS-PCR results and DNA sequencing findings was 99.4%. With culture-based phenotypic drug susceptibility testing as a reference standard, the sensitivity and specificity of the newly developed MAS-PCR assay for drug resistance-related genetic mutation detection were determined to be 81.1% and 97.5% for INH, 93.0% and 98.9 % for RIF, and 54.5% and 68.0 % for ethambutol. MAS-PCR provides a rapid, potentially more cost-effective, method of detecting multidrug-resistant TB. PMID- 16243478 TI - Evaluation of the BD Phoenix Automated Microbiology System SMIC/ID panel for identification and antimicrobial susceptibility testing of Streptococcus spp. AB - The BD Phoenix Automated Microbiology System SMIC/ID panel was evaluated for identification and antimicrobial susceptibility testing (AST) of various streptococci. A group of 97 consecutive clinical isolates of Streptococcus pneumoniae, 23 Streptococcus pyogenes, 24 Streptococcus agalactiae, and 34 viridans streptococci were collected and comparisons made with routine manual methods used in the clinical microbiology laboratory. Overall, in 162 (91%) of 178 isolates, Phoenix identification results demonstrated agreement. For AST results for the 162 isolates that demonstrated identification concordance, the overall essential agreement rate was 98.5%; the category agreement was 94.9%; and the very major error, major error, and minor error rates were 0%, 0.15%, and 5.8%, respectively. Although relatively high minor error rates were observed with S. pneumoniae and beta-lactams, 79.2% of the 77 minor errors were the result of a single log(2) dilution difference. The Phoenix SMIC/ID panel performed favorably and demonstrated the advantages of automation and simple methodology. PMID- 16243479 TI - Reconstituted human corneal epithelium: a new alternative to the Draize eye test for the assessment of the eye irritation potential of chemicals and cosmetic products. AB - The aim of this study was to evaluate the interest of a new three-dimensional epithelial model cultivated from human corneal cells to replace animal testing in the assessment of eye tolerance. To this end, 65 formulated cosmetic products and 36 chemicals were tested by means of this in vitro model using a simplified toxicokinetic approach. The chemicals were selected from the ECETOC data bank and the EC/HO International validation study list. Very satisfactory results were obtained in terms of concordance with the Draize test data for the formulated cosmetic products. Moreover, the response of the corneal model appeared predictive of human ocular response clinically observed by ophthalmologists. The in vitro scores for the chemicals tested strongly correlated with their respective scores in vivo. For all the compounds tested, the response of the corneal model to irritants was similar regardless of their chemical structure, suggesting a good robustness of the prediction model proposed. We concluded that this new three-dimensional epithelial model, developed from human corneal cells, could be promising for the prediction of eye irritation induced by chemicals and complex formulated products, and that these two types of materials should be tested using a similar protocol. A simple shortening of the exposure period was required for the chemicals assumed to be more aggressively irritant to the epithelial tissues than the cosmetic formulae. PMID- 16243480 TI - Sleep and body temperature responses in an acute viral infection model are altered in interferon type I receptor-deficient mice. AB - Type I interferons (IFNs) include IFNalpha and IFNbeta, both of which are elevated in acute viral infections and both of which have been shown to induce symptoms such as fever and somnolence when administered in pharmacological doses. To investigate the role of type I IFNs in mediation of acute respiratory viral symptoms we examined sleep and body temperature responses in mice with a targeted mutation of the IFN receptor type I (IFN-RI knockouts). IFN-RI knockouts (KOs) or wild-type 129 SvEv controls were challenged intratracheally (IT) with combined poly[rI.rC] (synthetic double-stranded RNA) and IFNgamma, a model that simulates an acute viral infection with respect to body temperature and locomotor activity responses. Control mice of both strains were treated with IT IFNgamma alone. Hypothermic responses to IT poly[rI.rC]/IFNgamma were more exaggerated in the IFN RI KO mice than in wild-type. The non-rapid eye movement sleep (NREMS) response to IT poly[rI.rC]/IFNgamma was increased earlier in the IFN-RI KO mice than in wild-type, though the total time spent in NREMS was reduced in the KOs compared to wild-type and the return to baseline NREMS was faster in the KOs. The quality of NREMS also was altered more extensively in the wild-type than in the KO mice. Spontaneous rapid eye movement sleep (REMS) was suppressed in IFN-RI KOs as previously reported, but was not substantially altered in either mouse strain by IT poly[rI.rC]/IFNgamma challenge. Our results implicate type I IFNs as inhibitors of the hypothermic response and enhancers of the NREMS response to IT poly[rI.rC]/IFNgamma, a model of acute viral infection. PMID- 16243481 TI - Rapid action of pesticides on cytosolic calcium concentrations in cultures of human umbilical vein endothelial cells. AB - Persistent metabolites of pesticides such as p,p'-DDE, at environmentally relevant concentrations, have been shown to have a rapid effect on intracellular calcium [Ca2+]i concentrations in human granulosa-lutein cells. Since endocrine disrupting substances can be transferred from the maternal circulation to the fetus the present study examined whether the pesticides, kepone, o,p-DDE, p,p' DDE and methoxychlor, could alter cytoplasmic calcium [Ca2+]cyt concentrations in human umbilical vein endothelial (HUVE) cells. Cultured HUVE cells were loaded with Fura-2 AM and changes in [Ca2+]cyt of single cells were studied using a dynamic digital Ca2+ imaging system. Kepone and methoxychlor consistently increased [Ca2+]cyt concentrations, similar to the effects of estradiol and progesterone. p,p'-DDE increased [Ca2+]cyt concentrations in 80% of experiments whereas o,p-DDE stimulated its increases in 42%. Estrone, estriol, pregnenolone and cortisol were not effective. These results demonstrate that pesticides can have a rapid effect on HUVE cells probably through a nongenomic mechanism of action. PMID- 16243482 TI - Investigation of anti-hyaluronidase treatment on vocal fold wound healing. AB - SUMMARY: Phytochemical constituents of medicinal plants demonstrate inhibition of tissue and bacterial hyaluronidase. Echinacoside is a caffeoyl conjugate of Echinacea with known anti-hyaluronidase properties. The purpose of this study was to investigate the wound healing effects of Echinacea on vocal fold wound healing and functional voice outcomes. Pig animal model. METHODS: Vocal fold injury was induced in 18 pigs by unilateral vocal fold stripping. The uninjured vocal fold served as control. Three groups of six pigs randomly received a topical application of 300, 600, or 1,200 mg of standardized Echinacea on the injured side. Animals were euthanized after 3, 10, and 15 days of wound healing. Phonation threshold pressure and vocal economy measurements were obtained from excised larynges. Treatment outcomes were examined by comparing the animals receiving treatment with a set of 19 untreated and 5 historical controls. Treatment effects on wound healing were evaluated by histologic staining for hyaluronan and collagen. Treated larynges revealed improved vocal economy and phonation threshold pressure compared with untreated larynges. Histologically, treated vocal folds revealed stable hyaluronan content and no significant accumulation of collagen compared with control. Findings provide a favorable outcome of anti-hyaluronidase treatment on acute vocal fold wound healing and functional measures of voice. PMID- 16243483 TI - Normative standards for vocal tract dimensions by race as measured by acoustic pharyngometry. AB - SUMMARY: Acoustic pharyngometry evaluates the geometry of the vocal tract with acoustic reflections and provides information about vocal tract cross-sectional area and volume from lip to the glottis. Variations in vocal tract diameters are needed for speech scientists to validate various acoustic models and for medical professionals since the advent of endoscopic surgical techniques. Race is known to be one of the most important factors affecting the oral and nasal structures. This study compared vocal tract dimensions of White American, African American, and Chinese male and female speakers. One hundred and twenty healthy adult subjects with equal numbers of men and women were divided among three races. Subjects were controlled for age, gender, height, and weight. Six dimensional parameters of the speakers' vocal tract cavities were measured with acoustic reflection technology (AR). Significant gender and race main effects were found in certain vocal tract dimensions. The findings of this study now provide speech scientists, speech-language pathologists, and other health professionals with a new anatomical database of vocal tract variations for adult speakers from three different races. PMID- 16243484 TI - Autoimmunity and clinical course in children with type 1, type 2, and type 1.5 diabetes. AB - AIMS: Both type 1 (T1D) and type 2 diabetes (T2D) are increasing in incidence in children; often an admixture of T1D and T2D features are present at diagnosis. We examined the relationship between diabetes autoantibodies (DAA), human leukocyte antigen (HLA), and clinical course in subjects grouped by clinical diabetes type. METHODS: Subjects 8-18 years old with T1D, T2D, and mixed clinical features (T1.5D), were studied at diagnosis. DAA were measured in all subjects; a subset of subjects underwent HLA genotyping. Clinical course was followed in 84% of subjects for 47.9+8.7 months. RESULTS: Eighty-nine percent of T1.5D subjects were positive for at least one DAA; 88% of HLA-typed subjects had risk HLA genotypes. Two subjects initially treated with oral agents were subsequently treated with insulin (50%); one had risk HLA, and the other was DAA positive. Thirty-three percent of T2D subjects were DAA positive and 93% were treated with oral agents at diagnosis. Three subjects were subsequently treated with insulin (21%); of these, two were DAA positive, and one had risk HLA. No subject who remained on diet therapy or oral agents had a combination of DAA-positivity and risk HLA genotype. CONCLUSIONS: Children clinically classified with T1.5D or T2D have a high frequency of autoimmune markers and T1D-associated HLA alleles which appears to indicate a more aggressive diabetes disease process, as has been shown for DAA positive adults with phenotypic T2D. PMID- 16243485 TI - Increased levels of chemokine receptor CXCR3 and chemokines IP-10 and MIG in patients with primary biliary cirrhosis and their first degree relatives. AB - Infiltrating memory T cells play an important role in the destruction of the biliary tract in primary biliary cirrhosis (PBC) and inflammatory chemokines control lymphocyte traffic through their interactions with T cell chemokine receptors. In the present study, we measured plasma levels of chemokines interferon-gamma-inducible protein-10 (IP-10) and monokine induced by gamma interferon (MIG), and also studied the expression of CXCR3 chemokine receptors in 105 subjects, including 53 patients with PBC, 26 first degree relatives and 26 healthy controls. Interestingly, plasma IP-10 and MIG levels in PBC were increased significantly compared to controls and appeared to increase with disease progression. By immunohistochemistry, IP-10 and MIG expressions were evident in the portal areas in PBC. Further, the frequency of CXCR3-expressing cells in peripheral blood was also significantly higher in PBC, and CXCR3 positive cells were also found in the portal areas of diseased livers, primarily on CD4+ cells. Finally, the daughters and sisters of PBC patients also demonstrated increased plasma levels of IP-10 and MIG, but, in contrast, displayed normal frequency of CXCR3+ expressing peripheral blood lymphocytes. Our data imply a role for specific chemokine-chemokine receptor interactions in the pathogenesis of PBC and also highlight the familial risk factor. PMID- 16243486 TI - The genetics of systemic lupus erythematosus. AB - For years the identification of candidate genes has been approached in various ways. The latest technology allows for whole genome SNP analysis in genetic association studies. These studies pose new challenges but also new possibilities. I hereby review what is known to date on lupus genetics and what challenges we are to overcome with the new available methods. PMID- 16243487 TI - IPEX and FOXP3: clinical and research perspectives. AB - Immunodysregulation, Polyendocrinopathy, Enteropathy, X-linked (IPEX) syndrome is a rare inborn error of immune regulation characterized by the early onset of one or more autoimmune diseases in boys. IPEX is caused by mutations in FOXP3, and is thus the homologue of the scurfy mutant mouse. The gene product, Scurfin, is required for the development of CD4+CD25+ T regulatory cells. In the absence of T regulatory cells, activated CD4+ T cells instigate multi-organ damage resulting in type 1 diabetes, enteropathy, eczema, hypothyroidism, and other autoimmune disorders. While effective therapies are currently limited, studies in the scurfy mouse are revealing aspects of pathophysiology and genetics that will lead to novel approaches for treating IPEX and other autoimmune disorders. Females carrying Foxp3 mutations are unaffected. In new experiments we show that female scurfy mice that are also heterozygous in trans for the X-linked recessive common gamma chain knockout contract autoimmune disease, proving that murine Foxp3 is subject to X-inactivation and providing an example of gene-gene interaction causing autoimmune disease in females. One explanation for the lesser disease severity in these females is proposed. PMID- 16243488 TI - LRRK1 protein kinase activity is stimulated upon binding of GTP to its Roc domain. AB - Human leucine-rich repeat kinase 1 (LRRK1) is a multi-domain protein of unknown function belonging to the ROCO family of complex proteins. Here, we report the molecular characterization of human LRRK1 and show, for the first time, that LRRK1 is both a functional protein kinase and a GDP/GTP-binding protein. Binding of GTP to LRRK1 is specific, requires the GTPase-like Roc domain, and leads to a stimulation of LRRK1 kinase activity. LRRK1 is the first example of a GTP regulated protein kinase harboring both the kinase effector domain and the GTP binding regulatory domain. Hence, we propose a model in which LRRK1 cycles between a GTP-bound active and a GDP-bound inactive state. Moreover, we mutated LRRK1 to mimic mutations previously identified in LRRK2/dardarin, the only human paralogue of LRRK1, that have been linked to autosomal-dominant parkinsonism. We demonstrate that three of four mutations analyzed significantly downregulate LRRK1 kinase activity. Ultimately, the results presented for LRRK1 may contribute to the elucidation of LRRK2's role in the pathogenesis of Parkinson's disease. PMID- 16243489 TI - Differences in performance and learning proficiency on the Wisconsin Card Sorting Test in schizophrenia: do they reflect distinct neurocognitive subtypes with distinct functional profiles? AB - The aim of this study was two-fold: 1) to determine whether a priori subtyping of patients with schizophrenia based on both overall performance on a measure of executive-function, the Wisconsin Card Sorting Test (WCST), and ability to learn the task with expanded instruction, could be confirmed with other, independent measures of executive-function and learning, and, if so, 2) to determine whether these groups have different neurocognitive profiles and show differences in functional capacity. Fifty-four outpatients with schizophrenia were divided by WCST performance into three groups: intact executive-function (n=28), "good learner" (n=13), and "poor learner" (n=13) groups. These groups were then assessed with a comprehensive neurocognitive test battery and a performance-based measure of functional status, the UCSD Performance-based Skills Assessment (UPSA). The WCST-intact subgroup performed significantly better than other groups on a second measure of executive-function and in working memory, and speeded motor sequencing. Impaired WCST patients who benefited from expanded WCST instruction ("good learners") also showed better performance than patients who did not benefit from instruction ("poor-learners") on a second measure of learning, as well as on a measure of auditory divided attention. The intact WCST subgroup had greater functional capacity than either "strong" or "poor" learners. These subtypes may have implications for response to behavioral treatment interventions. PMID- 16243490 TI - Reduced perception of the motion-induced blindness illusion in schizophrenia. AB - Motion-induced blindness (MIB) occurs when target stimuli are presented together with a moving distractor pattern. Most observers experience the targets disappearing and reappearing repeatedly for periods of up to several seconds. MIB can be viewed as a striking marker for the organization of cognitive functioning. In the present study, MIB rates and durations were assessed in 34 schizophrenia spectrum disorder patients and matched controls. The results showed that positive symptoms and excitement enhanced MIB, whereas depression and negative symptoms attenuated the illusion. MIB was more frequently found in normal subjects. The results remained consistent after adjusting for reaction time and error rates. Hence, MIB may provide a valid and reliable measure of cognitive organization in schizophrenia. PMID- 16243491 TI - Clozapine as a short-term antipsychotic medication in patients with schizophrenia. PMID- 16243492 TI - Management of the febrile neutropenic patient. AB - Fever of unknown origin in oncological patients is a frequent problem throughout the world. The microbiology of infections in these patients can vary widely. Gram negative bacteria were more prevalent in early trials, but Gram-positive organisms have become increasingly common since the mid 1980s. However, Gram negative microorganisms appear to be resurging. Equally important changes have occurred in the antimicrobial susceptibility of infective pathogens, most importantly methicillin-resistant Staphylococcus aureus, coagulase-negative staphylococci, vancomycin-resistant enterococci, viridans group streptococci, ciprofloxacin-resistant Escherichia coli and Pseudomonas aeruginosa. Current management strategies for febrile neutropenic patients emphasize risk assessment and the suitability of individual patients for outpatient versus hospital treatment and for oral versus parenteral therapy. Among the new determinants of infection risk, the most important are the severity and duration of neutropenia. Additional significant issues include: the selection of monotherapy versus combination therapy; and prophylaxis, which involves, among other strategies, quinolone use, prevention of fungal and viral infections, surveillance cultures, prevention of catheter-related infections, and vaccines. With relation to the consensus document, it should clearly define fever and neutropenia, and rank the strength of recommendations and the quality of the evidence on which they are based. Finally, the document should provide a detailed, stepwise management algorithm, addressing the initial empirical antimicrobial therapy and the antimicrobial therapy on days 3-5 and days 5-7 of therapy. PMID- 16243493 TI - Red cell membrane omega-3 fatty acids are decreased in nondepressed patients with social anxiety disorder. AB - The "phospholipid hypothesis" attributes a pathophysiologic role to the polyunsaturated fatty acid (PUFA) composition of phospholipids in depression. The aim of the present study was to determine whether the hypothesis is relevant to social anxiety disorder (SAD). The study sample consisted of 27 untreated, nondepressed patients with SAD (DSM-IV) and 22 controls. Severity of SAD was assessed with the Liebowitz Social Anxiety Scale (LSAS). Erythrocyte PUFA concentrations were measured by gas-liquid chromatography. Concentrations of most n-3 PUFAs were lower in the patients: 18:3n-3 by 32% (p < 0.002), 20:3n-3 by 34%, 20:5n-3 by 36% (all p < 0.001) and 22:6n-3 by 18% (p = 0.002). No significant differences were observed in other fatty acids. Significant inverse correlations were obtained between levels of n-3 PUFAs and LSAS scores. In conclusion, the phospholipid hypothesis may apply to SAD, thereby opening new therapeutic options. The robust relationship between low erythrocyte n-3 PUFA concentrations and SAD justifies exploration of relevant neuropathophysiological mechanisms. PMID- 16243494 TI - Prefrontal abnormalities in patients with simple schizophrenia: structural and functional brain-imaging studies in five cases. AB - Simple schizophrenia is an uncommon disorder with unknown pathophysiology, and its position in the current diagnostic system is ambiguous. Brain-imaging studies may help to elucidate its pathophysiology. Five patients fulfilling both ICD-10 criteria for simple schizophrenia and DSM-IV criteria for simple deteriorative disorder underwent computed tomography, magnetic resonance imaging, and single photon emission computed tomography. These scans were assessed individually by visual inspection as well as automatically by comparison with scans in normal controls or other schizophrenia subtype patients using voxel-based image analyses. Three of the five simple schizophrenia patients had findings of atrophy and reduced cerebral perfusion in the frontal areas. Voxel-based analyses also showed prefrontal grey matter deficits and hypoperfusion in simple schizophrenia patients compared with the controls. Although this study is limited by the small number of patients with simple schizophrenia, the results suggest that simple schizophrenia, or at least this subpopulation, may have rather homogeneous morphological and functional deficits in the prefrontal cortex. It is also suggested that simple schizophrenia may occupy an extreme position of the schizophrenic continuum where the prefrontal deficits and negative symptoms are most purely manifested. PMID- 16243495 TI - Attention and selection for predictive smooth pursuit eye movements. AB - Humans cannot typically produce smooth eye movements in the absence of a moving stimulus. However, they can produce predictive smooth eye movements if they expect a target of a known velocity to reappear. Here, we observed that participants could extract velocity information from two simultaneously presented moving targets in order to produce a subsequent predictive smooth eye movement for one of the two targets. Subjects fixated a stationary cross during the presentation of two targets, moving rightward at different velocities. In the next presentation, a single target was presented, which participants tracked with their eyes. A static cue, presented 700 ms before the moving target, indicated which of the two targets would be presented. Predictive eye movements were of an appropriate velocity, even when participants did not know in advance which of the two targets would subsequently be cued. However, the scaling of predictive eye velocity was marginally less accurate in this divided attention condition than when participants knew the identity of the cued target in advance, or a single target was presented during fixation. In a second experiment, we found that the velocity cued on the previous trial had a greater effect than the uncued velocity on the current trial. The negligible effect of the uncued velocity indicates that participants were extremely effective at selectively reproducing one of two recently viewed velocities. However, other influences, such as past history, also affected predictive smooth eye movements. PMID- 16243496 TI - In situ forming microparticle system for controlled delivery of leuprolide acetate: influence of the formulation and processing parameters. AB - The objective of present study was to control the delivery of leuprolide acetate using in situ forming microparticle (ISM) systems. A solution of leuprolide acetate and poly(lactide-co-glycolide) (PLGA RG 503H) or poly(lactide) (PLA R 202H) in N-methyl-2-pyrrolideone (NMP) was emulsified into an external oil phase using a two-syringe/connector system. After injection into an aqueous environment, NMP diffusion led to polymer precipitation and microparticle formation in situ. ISM-systems were characterized with respect to particle morphology and the influence of formulation and processing parameters on the in vitro release. ISM from RG 503H showed a high initial release (approximately 40%), which could be attributed to the high porosity of microparticles. The initial release could be reduced by increasing the polymer concentration, increasing the amount and viscosity of the oil phase, and decreasing the drug loading. ISM-systems from R 202H had a much lower initial release (approximately 9%) compared to that from RG 503H, which was followed by a slow and continuous drug release. In comparison to conventional microparticles prepared by a solvent evaporation method, ISM from R 202H showed a lower initial release and a more linear continuous release. PMID- 16243497 TI - [Importance of anamnestic criteria of perinatal infections for the diagnosis of early bacterial maternofetal infection]. PMID- 16243499 TI - Does training of respiratory muscles affect exercise performance in healthy subjects? PMID- 16243498 TI - Control of mild to moderate asthma over 1-year with the combination of salmeterol and fluticasone propionate. AB - OBJECTIVE: The aim of this study was to assess asthma control using salmeterol plus fluticasone propionate (FP) in combination (SFC) versus salmeterol or FP as monotherapy in patients with mild to moderate asthma. METHODS: In this randomised, double-blind, parallel-group study, 322 symptomatic patients were recruited, of which 282 were randomised to receive either salmeterol (50 microg), FP (250 microg), or SFC (50 microg/250 microg), via a single Diskus inhaler twice daily for 12 months. Outcome variables included the number of patients requiring an increase in study medication and the number experiencing 2 exacerbations during the 12-month treatment period. Airway hyper-responsiveness (AHR) and lung function tests were performed at clinic visits. Peak expiratory flow, rescue medication use, symptom scores and adverse events were recorded in diary cards. RESULTS: Fewer patients required an increase in study medication with SFC (10.5%) than with either FP (34.8%) or salmeterol (61.1%) (P<0.001). Significantly fewer patients experienced 2 exacerbations with SFC (4.2%), compared with FP (17.4%; P<0.01) or salmeterol (40%; P<0.001). SFC improved AHR to a significantly greater extent than FP (methacholine PC20=1.8 mg/ml vs. 1.1 mg/ml; P<0.05) or salmeterol (methacholine PC20=1.8 mg/ml vs. 0.7 mg/ml; P<0.001). CONCLUSIONS: The protection against exacerbations may be attributed to better control of inflammation, AHR and lung function parameters achieved with salmeterol and FP in combination, compared with either treatment alone. PMID- 16243500 TI - The effect of massive weight loss on pulmonary function of morbid obese patients. AB - STUDY OBJECTIVES: To test if morbid obesity causes pulmonary function changes and if massive weight loss have effect on pulmonary function (especially in subjects with BMI>or=60 kg/m(2)). PARTICIPANTS: Thirty-nine morbid obese subjects before and after massive weight loss. MEASUREMENTS AND RESULTS: Patients had baseline BMI>or=40 kg/m(2), pulmonary function test (PFT) before and after surgery for gastric volume reduction and massive weight loss, and presented no complaints unrelated to obesity. Based on initial BMI, the patients were divided in groups A (BMI 40-59.9 kg/m(2)) and B (BMI>or=60 kg/m(2)). Initially, group A (n=28) had normal PFT, however group B (n=11) presented FVC and FEV(1) measurements in the lowest limit of normality (with normal FEV(1)/FVC), significantly different from group A. After massive weight loss, the group B compared to A had a significant improvement in FVC (23.7% vs. 9.7%, P=0.012) and FEV(1) (25.6% vs. 9.1%, P=0.006); thus the initial difference in FVC and FEV(1) between groups no longer existed after weight loss. CONCLUSIONS: These results point out that the severe morbid obesity (BMI>or=60 kg/m(2)) may lead to pulmonary function impairment and presents more prominent pulmonary function gain after massive weight reduction. The possible clinical implications of these results are that PFT abnormalities in subjects with BMI<60 kg/m(2) should probably be interpreted as consequence of intrinsic respiratory disease and that severe morbid obese patients may be encouraged to lose weight to improve their pulmonary function, especially those with concomitant pulmonary disorders. PMID- 16243501 TI - On the cutting edge of clinical pulmonary medicine in COPD. PMID- 16243502 TI - Regulation of temporal identities during Drosophila neuroblast lineage development. AB - One of the major goals of neurobiology is to describe, in molecular terms, how a neural progenitor cell can generate an ordered series of uniquely fated neurons and glia. It has become clear that many, or all, neural-subtype identities can be linked to sequentially changing regulatory programs within neural precursors. Recent studies shed light on regulatory inputs and timing mechanisms that generate temporally defined cell identities, and new contributions are beginning to establish a link between the temporal network and cell function. PMID- 16243504 TI - Heterotrimeric G proteins and regulation of size asymmetry during cell division. AB - The generation of daughter cells of different fate and size depends on the orientation, positioning and morphology of the mitotic spindle. In both C. elegans and Drosophila, heterotrimeric G proteins have emerged as central and conserved regulators of this process. Although the same molecular players are involved in worms and flies, there are clear differences in the mechanisms used. Interestingly, recent work in mammalian cells suggests that heterotrimeric G proteins may control spindle positioning in higher organisms during symmetric and asymmetric cell divisions. PMID- 16243506 TI - Symmetric versus asymmetric cell division during neurogenesis in the developing vertebrate central nervous system. AB - The type and number of cell divisions of neuronal progenitors determine the number of neurons generated during the development of the vertebrate central nervous system. Over the past several years, there has been substantial progress in characterizing the various kinds of neuronal progenitors and the types of symmetric and asymmetric divisions they undergo. The understanding of the cell biological basis of symmetric versus asymmetric progenitor cell division has been consolidated, and the molecular machinery controlling these divisions is beginning to be unravelled. Other recent advances include comparative studies of brain development in rodents and primates, as well as the identification of gene mutations in humans that affect the balance between the various types of cell division of neuronal progenitors. PMID- 16243508 TI - Effects of different phytosterol analogs on colonic mucosal cell proliferation in hamsters. AB - OBJECTIVE: The aim of this study was to investigate the effects of different phytosterols and their analogs on colonic mucosal cell proliferation in hamsters. METHOD: Hamsters (n=70) were randomly assigned to seven groups after a 2-week acclimation and fed the experimental diet for 5 weeks. Diets included (i) the semipurified diet with no cholesterol (Con), (ii) the Con diet plus 0.25% cholesterol (Ch-con), or the Ch-con diet with (iii) 1% phytosterols (Ste), (iv) 1% phytostanols (Sta), (v) 1.76% sterol esters (esterified to fish oil, SteF), (vi) 0.71% stanol esters (esterified to ascorbic acid [disodium ascorbyl phytostanol phosphate, FM-VP4], 0.7% StaA) and (vii) 1.43% stanol esters (1.4% StaA), respectively. After 5 weeks on experimental diet, hamsters were sacrificed, and colons were collected. Colonic mucosal cell proliferation was measured by immunohistochemistry using monoclonal antibodies against antigen Ki 67. RESULTS: Colonic mucosal cell proliferation was 21.4% (P<.01) lower in the 0.7%, but not 1.4%, StaA relative to the Ch-con group. In addition, a lower ( 13.9%) cell proliferation was observed in the SteF group in comparison to the Ch con group; however, this difference achieved only a borderline level of statistical significance (P=.069). No differences were observed between Con and Ch-con, as well as among Ste, Sta, 1.4% StaA and Ch-con treatments. CONCLUSION: Plant stanols esterified to ascorbic acid may possess anticarcinogenic properties in the colon by suppressing colonic mucosa cell proliferation; however, this effect was not observed with free plant sterols or stanols. PMID- 16243507 TI - Life in the balance: how BH3-only proteins induce apoptosis. AB - BH3-only members of the Bcl-2 intracellular protein family, which include Bim, Bmf, Bik, Bad, Bid, Puma, Noxa and Hrk, mediate many developmentally programmed and induced cytotoxic signals. They have key roles in development, tissue homeostasis, immunity and tumor suppression, and compounds mimicking them are promising anti-cancer agents. Their activity is normally constrained by transcriptional and/or diverse post-transcriptional controls. When activated, these death ligands engage pro-survival Bcl-2-like proteins via the BH3 domain, inactivating their function. Bim and Puma bind all the pro-survival proteins, whereas others, such as Noxa and Bad, engage distinct subsets and exhibit complementary killing. Hence, multiple pro-survival proteins must be inactivated to unleash Bax and Bak, which drive apoptosis. Whether certain BH3-only proteins also directly activate Bax/Bak remains controversial. PMID- 16243509 TI - Prolonged increase in dietary phosphate intake alters bone mineralization in adult male rats. AB - Excessive intake of dietary phosphate without the company of calcium causes serum parathyroid hormone (s-PTH) concentration to rise. We investigated the effect of a modest but prolonged increase in dietary intake of inorganic phosphate on the bone quantitative factors of mature male rats. Twenty Wistar rats were divided into two groups and fed a high-phosphate diet (1.2% phosphate) or a control diet (0.6% phosphate) for 8 weeks. In the beginning and at the end of the study period, femur and lumbar bone mineral density (BMD), bone mineral content and area were measured using DXA, s-PTH was analyzed from the blood sample, and after sacrifice, right femur was cut loose and processed into paraffin cuts. Bone diameter, inner diameter and cortical width was measured from the hematoxylin- and eosin-dyed femur cuts. Tibias were degraded and calcium and phosphate content was analyzed by inductively coupled plasma-mass spectrometer. Femoral BMD increased significantly more in the control group than in the phosphate group (P=.005). Lumbar BMD values decreased in both groups, and the fall was greater in the control group (P=.007). The phosphate group had significantly higher s-PTH values (P=.0135). Femoral histomorphometric values or tibial mineral contents did not differ between groups. In conclusion, increase in dietary phosphate intake caused s-PTH to rise and hindered mineral deposition into cortical bone, leading to lower BMD. The effect on trabecular bone was opposing as mineral loss was less in the lumbar spine of phosphate group animals. These results are in concurrence with the data stating that skeletal response to PTH is complex and site dependent. PMID- 16243510 TI - Electrochemical microdevice with separable electrode and antibody chips for simultaneous detection of pepsinogens 1 and 2. AB - An electrochemical microdevice with separable electrode and antibody chips has been developed and applied to detect atrophic gastritis-related proteins, pepsinogen 1 (PG1) and pepsinogen 2 (PG2), based on sandwich-type enzyme-linked immunosorbent assays (ELISAs) with horseradish peroxidase (HRP)-labeled antibody. To fabricate the electrochemical device for simultaneous analysis of several proteins, the electrode chip with eight electrode elements was assembled along with an antibody chip with eight cavities containing immobilized anti-PG1 or anti PG2. The immunoreactions occurring in the cavities of the device were detected simultaneously by amperometry. The labeled HRP in the cavity in the presence of hydrogen peroxide catalyzed the oxidation of ferrocenemethanol (FMA) to FMA+, which was detected electrochemically by the electrode chip. The amperometric responses of respective cavities in the device increased with increasing concentration of PG1 or PG2 of 0-50 ng/ml, ensuring the simultaneous detection of PG1 and PG2. The detection limits for both PG1 and PG2 were 0.6 ng/ml (S/N=2). The electrode chip was recovered easily by disassembling the electrochemical device; thereby, it was used repeatedly, whereas the antibody chip was discarded. No marked decrease in electrochemical responses was detected after repeated use. Reuse of the electrode chip is beneficial to reduce costs of protein analysis. PMID- 16243511 TI - Development of a combined setup for simultaneous detection of total and glycated haemoglobin content in blood samples. AB - An original setup for analysis of glycated haemoglobin (HbA1c) in blood is reported. The construction employed a combination of the piezoelectric biosensor for glycated haemoglobin and the flow-through photometric sensor for total haemoglobin (Hb). The modification of gold electrodes with 3-aminophenylboronic acid (APBA) as a specific ligand was studied; the chemisorbed conjugate of APBA with a long-chain thiocompound provided the best affinity for HbA1c. The effect of various operating parameters, such as flow rate and instrumental setup, was optimised. The total haemoglobin content was analysed as absorbance of the haemoglobin-cyanide derivative at 540 nm. Only one standard (calibrator) diluted in various ratio was necessary for calibration and 1 microl of blood was sufficient for analysis. The full range of HbA1c content (4-15%) in blood can be analysed; the working ranges of total and glycated haemoglobin were 50-2000 and 10-90 microg/ml, respectively. The developed method was successfully evaluated on blood samples collected from diabetics. PMID- 16243512 TI - Catalytic antibodies and their applications. AB - Catalytic antibodies (CAbs) occur naturally in healthy individuals where they may form part of the innate immune system, but are preferentially found in those with autoimmune disease. CAbs can also be artificially engineered or elicited by immunizations. Their mechanisms of action include nucleophilic catalysis, induction of conformational strain, coordination with metal ions, and stabilization of transition states. Recent applications of CAbs with clinical significance include the conversion of cocaine to a non-psychoactive form, the degradation of nicotine, activation of prodrugs for targeted chemotherapy, protection from ultraviolet radiation, inhibition of HIV infectivity, and the destruction of aggregates of beta-amyloid implicated in Alzheimer's disease. Artificial CAbs are likely to find increasing applications in research, clinical medicine, diagnostics and manufacturing. PMID- 16243513 TI - Hyperinsulinaemia: a prospective risk factor for lethal clinical prostate cancer. AB - Previous studies have suggested that hyperinsulinaemia and other components of metabolic syndrome are risk factors for clinical prostate cancer. This prospective study tested the hypothesis that hyperinsulinaemia and other components of metabolic syndrome are risk factors for lethal clinical prostate cancer. The clinical, haemodynamic, anthropometric, metabolic and insulin profile at baseline in men who had died from clinical prostate cancer during follow-up was compared with the profile of men who were still alive at follow-up. If the hypothesis is true, men with an unfavourable prognosis would have a higher profile at baseline than those with a favourable prognosis. A total of 320 patients in whom clinical prostate cancer, stages T2-3, had been diagnosed were consecutively included in the study during 1995-2003. Height, body weight, waist measurement, hip measurement and blood pressure were determined. Body mass index and waist/hip ratio (WHR) were calculated. Blood samples were collected to determine triglycerides, total cholesterol, high-density lipoprotein (HDL) cholesterol, low-density lipoprotein (LDL)-cholesterol, uric acid, alanine aminotransferase and fasting plasma insulin level. The prostate gland volume was measured using transrectal ultrasound. The annual benign prostatic hyperplasia (BPH) growth rate was calculated. The diagnosis of prostate cancer was established using transrectal ultrasound-guided automatic needle biopsy of the prostate gland. All patients with clinical prostate cancer were followed up until their death or until the study was terminated on 31 December 2003. At follow-up, 54 patients had died from prostate cancer and 219 were still alive. The results showed that the men who died of clinical prostate cancer during the follow-up period were older (P < 0.001), had a larger prostate gland volume (P < 0.001), a faster BPH growth rate (P < 0.001), a higher prevalence of type 2 diabetes (P < 0.035) and treated hypertension (P < 0.023), a higher stage (P < 0.001) and grade (P = 0.028) of clinical prostate cancer, a higher prostate-specific antigen (PSA) level (P < 0.001) and a higher PSA density (P < 0.001) at baseline than men still alive with clinical prostate cancer at follow-up. These men also had a lower HDL cholesterol level (P = 0.027), a higher fasting plasma insulin level (P = 0.004), a higher WHR (P = 0.097) of borderline significance and a higher uric acid level (P = 0.079) of borderline significance. Eliminating the effect on mortality of higher stage and grade of the clinical prostate cancer and PSA at baseline, the following statistically significant correlations remained: a higher fasting plasma insulin level (P = 0.010) and a lower HDL-cholesterol level of borderline significance (P = 0.065). In conclusion, hyperinsulinaemia and five other previously established components of metabolic syndrome are shown to be prospective risk factors for deaths that can be ascribed to prostate cancer. These findings confirm previous study, which indicate that prostate cancer is a component of metabolic syndrome. Moreover, these data indicate that hyperinsulinaemia and other metabolic disorders precede deaths caused by prostate cancer. Thus, our data support the hypothesis that hyperinsulinaemia is a promoter of clinical prostate cancer. Furthermore, our data suggest that the insulin level could be used as a marker of prostate cancer prognosis and tumour aggressiveness, regardless of the patient's prostate cancer stage, cancer grade and PSA level. PMID- 16243514 TI - Preventing organ-specific chemotherapy toxicity. AB - Recent advances in treatment for pediatric cancers has increased overall survival rates. As more and more survive pediatric cancer, we continue to see the emergence of late effects of treatment within pediatric and the growing adult survivor population. The evaluation of late effects was initiated approximately two decades ago, and has become an extremely important facet of pediatric oncology. This review delves into several of the most serious organ-specific late effects of pediatric cancer treatment, outline what we know and what we do not currently understand about preventing or reducing them. Clinical and bench research are necessary to develop interventions that will avoid or mitigate late effects and improve the health of pediatric cancer survivors. PMID- 16243515 TI - Incidence of pleural mesothelioma in Liguria Region, Italy (1996-2002). AB - In this study, incidence of pleural malignant mesothelioma (PMM) in the Liguria Region (Italy) (approximately 1.6 million inhabitants), in the presence of asbestos exposure was investigated. New PMM cases recorded by the Mesothelioma Registry of Liguria, from 1996 to 2002 and interviews reported on a standardised questionnaire were analysed according to demographical and etiological characteristics. Nine hundred and forty five PMM cases were recorded (757 males and 188 females); the age standardised (European population) incidence rates per 100,000 were 8.51 and 1.43, respectively. The rates among the four provinces ranged between 1.18 and 13.7 for males and 0.68 and 1.44 for females. The questionnaire was evaluated for 786 PMM cases (or next-of-kin). Higher incidence rates were reported in the provinces with larger industrial and harbour areas, including shipyards (construction and repair), dockyards, building activities, chemical and heavy industrial activities. Asbestos exposure was unlikely or unknown for 57.5% females and 15% males. A major role of environmental asbestos exposure in the etiology of PMM is hypothesised for females and for a minor proportion of males. PMID- 16243516 TI - Risk of melanoma following adulthood cancer: a case-control study. AB - Melanoma is a severe skin cancer related to sun exposure. Whether this malignancy is linked to exposure to ionising radiation during adulthood is still controversial. This case-control study examined the risk of melanoma following treatment for an adulthood first malignant neoplasm (FMN). Cases were patients who presented with cutaneous melanoma after a first cancer in adulthood. Controls (3 per case) were patients free of melanoma, matched for age, duration of follow up since the FMN, type of FMN, and followed in the same institution. A total of 57 cases and 171 controls were included. In the final multivariate analysis, no risk of melanoma was associated with radiotherapy (odds ratio (OR) for 1 Gy = 1.01, 95% confidence interval (95%CI) 0.96-1.07) nor hormonotherapy, whereas chemotherapy use (OR = 2.3, 95%CI 0.93-5.6) and having a history of familial cancer (OR = 2.8, 95%CI 1.3-5.9) exhibited a nearly significant risk. In conclusion, unlike the evidence for risk of exposure to ionising radiation during childhood, we did not substantiate a risk for association of melanoma with exposure to ionising radiation during adulthood. The risk associated with chemotherapy should justify the implementation of skin surveillance for early detection of melanoma in these patients. PMID- 16243517 TI - Is there a common aetiology for certain childhood malignancies? Results of cross space-time clustering analyses. AB - We previously demonstrated significant space-time clustering amongst cases of childhood leukaemia (in particular acute lymphoblastic leukaemia (ALL)), central nervous system (CNS) tumour (especially astrocytoma), soft tissue sarcoma and Wilms' tumour. We hypothesised that there may be common aetiological mechanisms between some of these diagnostic groups. To test this hypothesis we analysed for cross-space-time clustering between these diagnostic groups, using population based data from north-west England. Data were examined by a second-order procedure based on K-functions. Reference points in time and space were dates and addresses at birth and diagnosis. The results showed statistically significant (P < 0.05) cross-clustering between cases of leukaemia and CNS tumour and between cases of ALL and astrocytoma. There was no statistically significant cross clustering of Wilms' tumours and soft tissue sarcomas with any other malignancy. In conclusion, these findings are consistent with common, possibly infectious, aetiological mechanisms for childhood leukaemia (particularly ALL) and CNS tumours (particularly astrocytoma). PMID- 16243519 TI - Cancer therapeutics: targeting the dark side of Myc. AB - The potent Myc oncoprotein plays a pivotal role as a regulator of tumorigenesis in numerous human cancers of diverse origin. Experimental evidence shows that inhibiting Myc significantly halts tumour cell growth and proliferation. This review summarises recent progress in understanding the function of Myc as a transcription factor, with emphasis on key protein interactions and target gene regulation. In addition, major advances in drug development aimed at eliminating Myc are described, including antisense and triple helix forming oligonucleotides, porphyrins and siRNA. Future anti-Myc strategies are also discussed that inhibit Myc at the level of expression and/or function. Targeting the dark side of Myc with novel therapeutic agents promises to have a profound impact in combating cancer. PMID- 16243521 TI - DT56a stimulates gender-specific human cultured bone cells in vitro. AB - DT56a found to have SERM-like properties is used for the treatment of menopausal symptoms and osteoporosis. In vivo experiments demonstrated that DT56a displayed selective estrogenic activity; it stimulated creatine kinase (CK) specific activity in the skeletal tissues but not on the uterus of ovariectomized rats. DT56a, when applied together with estradiol-17beta (E(2)), completely inhibited the E(2)-stimulated CK, as demonstrated by other SERMs. DT56a stimulated bone formation in a rat model as measured by histological and histomorphometrical parameters. In a clinical study, administration of DT56a (Femarelle) resulted in a considerable elevation of bone mineral density and relief of menopausal symptoms. The aim of the present study was to analyze the effects of DT56a in vitro on human-derived bone cultured osteoblasts (Ob), by measuring its effects, at different concentrations, on DNA synthesis, CK and alkaline phosphatase (ALP) specific activities as well as changes in intracellular [Ca(2+)](i) concentrations. DT56a stimulated CK and DNA synthesis in both pre- and post menopausal female Ob with maximal effect at 100 ng/ml for both age groups. In addition, DT56a stimulated ALP in Ob from both pre- and post-menopausal women with maximal effect at lower dose of 50 ng/ml, with higher response of pre menopausal cells. Raloxifene (Ral) inhibited all DT56a-stimulated changes in Ob from both age groups. DT56a, when given together with E(2), completely antagonized E(2)-stimulated effects demonstrating its nature as a phyto-SERM. DT56a also, dose dependency, stimulated the intracellular levels of [Ca(2+)](i) with maximal effect at 10 ng/ml. Male-derived Ob did not respond to DT56a in any parameter. In summary, DT56a stimulated sex-specifically female-derived Ob, indicating its unique nature compared to the compounds currently used for postmenopausal osteoporosis by being bone-forming and not only an anti-resorptive agent. PMID- 16243523 TI - An assessment of the toxicity of metals to Pseudomonas aeruginosa PU21 (Rip64). AB - The toxicity of Co(II), Mn(II), Cd(II), and Zn(II) for Pseudomonas aeruginosa PU21, a Hg(II)-hyperresistant strain containing the mercury resistance mer operon, was determined. The metal tolerance of PU21 was strongly influenced by environmental conditions (e.g., existing metal, medium composition). Dose response analysis on chronic and acute toxicity (e.g., EC(20), median effective dose EC(50), and slope factor B) of divalent cobalt, manganese, cadmium, and zinc cations in LB medium amended with citric acid phosphate buffered saline (CAPBS) suggested a toxicity series of Co > Mn approximately Zn > Cd for EC(50). In contrast, excluding the likely precipitate of Zn(II), the toxicity ranking in phosphate-buffered saline (PBS)-amended LB medium was Co > Cd > Mn. The metal toxicity in PBS, irrespective of metals, was greater than that in CAPBS. This might be attributed to the presence of citric acid in CAPBS as a chelating ligand donating electrons to hold free metals (e.g., Cd(2+), Zn(2+) tetrahedral ML(4) complex). The toxicity assessment established viable operation ranges (ca. 2000 nM) and versus kappa (Ki at kappa-receptors > 5000 nM) receptors. Molecular modelling carried out on the pair 3Aa/3Ba and on the 3Bh was consistent with the hypothesis that the two series of compounds 3A and 3B interact with the mu-receptor in very different ways. PMID- 16243531 TI - Histomorphometric analysis of glucocorticoid-induced osteoporosis. AB - Bone histomorphometry or quantitative histology consists of counting or measuring tissue components: cells, extracellular constituents and microarchitecture. Bone histomorphometry is the only method that allows the measurement of mineralization rate and the study of bone formation at three levels: cell, remodeling unit and tissue levels. It is a useful tool to explain the pathogenesis and cellular mechanisms of different metabolic bone diseases such as glucocorticoid-induced osteoporosis (GIO). Glucocorticoids (GC) affect calcium and bone metabolism at every level, but the main effect is the osteoblastic dysfunction. Concerning the bone formation, some histomorphometric studies have shown a depressed osteoblastic activity at a cell, bone remodeling unit, and tissue levels. In addition, there is evidence of a shortening of the period in which the osteoblasts work actively forming the bone matrix. This latter effect seems to occur after high cumulative doses of GC. With regard to the resorption, the results are still debated, but histomorphometric parameters seem to be increased in the majority of studies, at least in the first period of the GC treatment. From a structural point of view, GC seem to induce a thinning of the trabeculae without their perforation, which occurs only after high cumulative doses. Anti resorptive treatments, such as bisphosphonates, are able to counteract the negative effects of GC on bone. In particular, along with their active working period, they prolong the lifespan of osteoblasts and osteocytes. In addition, the anti-resorptive treatments seem to extend the time for secondary mineralization through a reduction of the Activation Frequency. The latter is an intriguing mechanism of bisphosphonates in GIO that needs further ad hoc investigations. PMID- 16243532 TI - Inactivation of vascular smooth muscle cells photosensitised by liposome delivered Zn(II)-phthalocyanine. AB - Photodynamic therapy is a promising approach for the prevention of arterial restenosis, which frequently occurs after balloon angioplasty, largely owing to abnormal proliferation of vascular smooth muscle cells (VSMC) and their migration from the media to the intima, where they originate intimal hyperplasia (IH). We investigated the efficacy of Zn(II)-phthalocyanine-photosensitised processes in promoting the inactivation of VSMC. This liposome delivered phthalocyanine is readily taken up by VSMC, largely partitions in the Golgi apparatus, and upon photoactivation causes >95% cell mortality using mild irradiation conditions (e.g. 5 min irradiation at 1 microM ZnPc). Cell death occurs through the parallel development of random necrotic and apoptotic processes. PMID- 16243533 TI - Effect of denaturation on the photochemistry of pyrimidine bases in isolated DNA. AB - The influence of denaturation on DNA photochemistry was studied by quantifying the yield of formation of all possible bipyrimidine photolesions within isolated genomic DNA samples exposed to UVC radiation. Effects of DNA melting was studied either by carrying out irradiation over a wide range of temperature (0-90 degrees C) or by decreasing the ionic strength of the solution at 30 degrees C. A first observation was a much larger decrease in the photoreactivity upon increasing the temperature in single-stranded than in double-stranded DNA. Secondly, formation of trans,syn cyclobutane dimers and, to a lesser extent, modification in the ratio between the yields of cyclobutane dimers and (6-4) photoproducts, were found to be other main features associated with denaturation. These results emphasize the modulating role of structure in the yield and nature of UV-induced DNA damage. PMID- 16243534 TI - Relationship between cytokine gene polymorphisms and graft-versus-host disease after allogeneic stem cell transplantation in a Brazilian population. AB - Graft-versus-host disease (GVHD) is the major complication of allogeneic hematopoietic stem cell transplantation (HSCT), and cytokines are recognized as important mediators of GVHD. Polymorphisms in the regulatory regions of several cytokine genes have been associated with a number of immune diseases as well as organ transplant complications. In this study we have investigated the role of tumor necrosis factor-alpha(-308), interleukin (IL)-6(-174), IL-10(-1082, -819, 592), Interferon-gamma(-874), and transforming growth factor-beta1(+869, +915) polymorphisms on HSCT outcome. Donor/recipient genotypes were analyzed by polymerase chain reaction with sequence specific primers (PCR-SSP). Although we have found a small number of low IL-6, a polymorphism at position -174 of the recipient and donor IL-6 gene was associated with the increased incidence of chronic GVHD. Therefore, this study emphasizes the probable potential role of genetic variability of donor and recipient in determining outcome after transplantation. PMID- 16243535 TI - Ginkgo biloba normalizes stress- and corticosterone-induced impairment of recall in rats. AB - Exposure to chronic restraint stress in rats and psychosocial stress in humans has been shown to alter cognitive functions such as learning and memory and has been linked to the pathophysiology of mood and anxiety disorders. Antianxiety or sedative agents used in the management of stress have several disadvantages and side effects. Therefore, in this study, we investigated efficacy of a natural medicine, the extract of Ginkgo biloba (EGB 761), in prevention and treatment of the post-stress memory dysfunctions. The results showed that chronic restraint stress (2h for 21 days) or an 'equivalent' dose of exogenous corticosterone (5 mg kg(-1)) decreased re-entry latencies in the passive avoidance situation showing thus impairment of recall. Preventive doses of EGB 761 (100 mg kg(-1)), given 30 min before each restraint stress episode or corticosterone injection, abolished cognitive deficits seen in unprotected rats. There was no influence of stress, corticosterone, and EGB 761 on the acquisition of conditioned avoidance responses (CARs). PMID- 16243536 TI - Piceatannol upregulates endothelial heme oxygenase-1 expression via novel protein kinase C and tyrosine kinase pathways. AB - Piceatannol is an anti-inflammatory and anti-proliferative plant-derived stilbene. Heme oxygenase-1 (HO-1) is a cytoprotective enzyme to activate by various phytochemicals. In this study, we examined the ability of piceatannol to upregulate HO-1 expression in endothelial cells. We found piceatannol at micromolar (10-50 microM) concentrations dramatically increased HO-1 protein levels in a time-dependent manner. Piceatannol was similarly potent in the induction of HO-1 as hemin, arsenate, and 15d-PGJ2, and was more potent than some other phytochemicals including curcumin, EGCG, baicalein, and quercetin. In contrast, the similar chemical structure compounds, trans-stilbene, stilbene oxide, and resveratrol had no HO-1-inducing effects, suggesting a critical role for the hydroxyl groups in HO-1 induction. No cytotoxicity and superoxide production was observed after 10-50 microM piceatannol treatments. Piceatannol mediated HO-1 induction was abrogated in the presence of N-acetylcysteine and glutathione, but not by other antioxidants. Induction of HO-1 by piceatannol was further enhanced by using buthionine sulfoximine. In addition, we determined that tyrosine kinase was involved in the induction of HO-1 by using tyrosine kinase inhibitors, herbimycin A, erbstatin, and genistein; in contrast, no significant changes in the pretreatment of PI3 kinase or MAP kinase inhibitors was determined. HO-1 induction was blocked by the protein kinase C inhibitors calphostin C, rottlerin, and long PMA pretreatment, whereas conventional PKC inhibitors, Go6976, and Ca2+ chelator BAPTA/AM, had no effect. Elevated HO-1 protein levels were associated with the inhibition of tumor necrosis factor-alpha (TNFalpha)-induced intercellular adhesion molecule-1 (ICAM-1) expression. Treating ECs with zinc protoporphyrin, an HO-1 inhibito blocked the anti inflammatory effect of piceatannol. In summary, this study identified piceatannol as a novel phytochemical inducer of HO-1 expression and identified the mechanisms involved in this process. PMID- 16243537 TI - Ventral midbrain glia express region-specific transcription factors and regulate dopaminergic neurogenesis through Wnt-5a secretion. AB - Glial cells have been classically described as supporting cells for neurons. Recently, additional roles during neural development have begun to emerge. Here, we report that ventral midbrain glia, including astrocytes and radial glia, are the source of signals required by neural precursors to acquire a dopaminergic phenotype. We found that ventral midbrain glia, but not cortical glia, secrete high levels of the glycolipoprotein Wnt-5a, express region-specific transcription factors such as Pax-2, En-1 and Otx-2 and increase the differentiation of cortical or ventral midbrain Nurr1 precursors into tyrosine hydroxylase-positive neurons. Moreover, blocking experiments using a Wnt-5a blocking antibody indicated that the effects of ventral midbrain glia on Nurr1-positive neural precursors are partially mediated by Wnt-5a. Thus, our results identify Wnt-5a as an important component of the dopaminergic inductive activity of the ventral midbrain glia. PMID- 16243538 TI - Expression, purification, and in vitro refolding of a humanized single-chain Fv antibody against human CTLA4 (CD152). AB - A human-derived single-chain Fv (scFv) antibody fragment specific against human CTLA4 (CD152) was produced at high level in Escherichia coli. The scFv gene was cloned from a phagemid to the expression vector pQE30 with a N-terminal 6His tag fused in-frame, and expressed as a 29 kDa protein in E. coli as inclusion bodies. The inclusion body of scFv was isolated from E. coli lysate, solubilized in 8M urea with 10mM dithiothreitol, and purified by ion-exchange chromatography. Method for in vitro refolding of the scFv was established. The effects of refolding buffer composition, protein concentration and temperature on the refolding yield were investigated. The protein was renatured finally by dialyzing against 3mM GSH, 1mM GSSG, 150 mM NaCl, 1M urea, and 50 mM Tris-Cl (pH 8.0) for 48 h at 4 degrees C, and then dialyzed against phosphate-buffered saline (pH 7.4) to remove remaining denaturant. This refolding protocol generated up to a 70% yield of soluble protein. Soluble scFv was characterized for its specific antigen binding activity by indirect cellular ELISA. The refolded scFv was functionally active and was able to bind specifically to CTLA4 (CD152). The epitopes recognized by refolded anti-CTLA4 scFv do not coincide with those epitopes recognized by CD80/CD86. PMID- 16243539 TI - Improved purification protocol for wild-type and mutant human foamy virus proteases. AB - Wild-type and an active site mutant (S25T) human foamy virus (HFV) proteases were expressed in fusion with maltose binding protein in Escherichia coli. The mutant enzyme contained a Ser to Thr mutation in the -Asp-Ser-Gly- active site triplet of the enzyme, which forms the "fireman's grip" between the two subunits of the homodimeric enzyme. The fusion proteins were purified by affinity chromatography on amylose resin, cleaved with factor Xa, and the processed enzymes were purified by gel filtration under denaturing condition. Refolding after purification resulted in active enzymes with comparable yields. Furthermore, both enzymes showed similar catalytic activities in an oligopeptide substrate representing an HFV Gag cleavage site. However, the S25T mutant showed increased stability in urea unfolding experiment, in a good agreement with the suggested role of the Thr residue of fireman's grip. PMID- 16243540 TI - Immunoreactive antigens of the outer membrane protein of Aeromonas hydrophila, isolated from goldfish, Carassius auratus (Linn.). AB - Aeromonas hydrophila causes disease under stress conditions or in concert with infection by other pathogens in goldfish. Sero-diagnostic and/or immunoprophylactic tools against Aeromonas infection in goldfish are not available so far. The present study was undertaken to fractionate and characterise the outer membrane proteins (OMP) of A. hydrophila and to identify suitable immunoreactive components. A total of 10 fractions were generated from crude OMP antigens upon gel permeation and subsequent ion-exchange chromatography. One of the fractionated antigens (GPID2), primarily a 57-kDa polypeptide, showed maximum sero-reactivity, even higher than the crude OMP. Suitability of GPID2 antigen for use in diagnostic preparations was assessed by dip-stick ELISA. In vitro goldfish lymphoproliferative ability of fractionated antigen, GPIID2 (primarily a 23-kDa polypeptide) was observed to be higher than all the fractionated antigens as well as crude OMP. It can be concluded that the 57 kDa and 23 kDa polypeptides of the OMP of A. hydrophila, possessing high immunoreactivity, should be given due attention while preparing immunodiagnostic and immunoprophylatic tools against Aeromonas infections in goldfish. PMID- 16243541 TI - Electromyographic analysis of joint-dependent global synkinesis in the upper limb of healthy adults: laterality of intensity and symmetry of spatial representation. AB - The intensity and spatial representation of electromyographical (EMG) activity were examined to characterize the effects of limb dominance and movement direction upon global synkinesis (GS). Twenty-two healthy young subjects (11 men, 11 women) with a mean age of 24.7 years participated in this study. Three trials of EMG activities from eight primary muscles in the unexercised limb were recorded when a maximal isometric contraction in various directions was performed by the shoulder, elbow, and wrist of the dominant and non-dominant upper limbs. The features of GS, including intensity and spatial representation, were quantified with standardized net excitation levels (SNE) and relative excitation (RE), respectively. Our data indicated that (1) GS intensity was strongly limb dependent with a larger SNE level arising when target joints of the non-dominant upper limb were active, (2) the GS intensity was more influenced by movement direction of the non-dominant limb than by that of the dominant limb, (3) the gradient change in GS intensity was observed bilaterally with a larger SNE level associated with contralateral movements of a proximal joint than a distal joint, and (4) GS spatial representations of the upper limbs were patterned and symmetrical, but seemly insensitive to movement direction. Laterality in GS intensity and structured GS spatial representation with symmetry could be a consequence of use-dependent hemispheric organization. PMID- 16243542 TI - Comparison of a piezoelectric contact sensor and an accelerometer for examining mechanomyographic amplitude and mean power frequency versus torque relationships during isokinetic and isometric muscle actions of the biceps brachii. AB - The purpose of this study was to compare a piezoelectric contact sensor with an accelerometer for measuring the mechanomyographic (MMG) signal from the biceps brachii during submaximal to maximal isokinetic and isometric forearm flexion muscle actions. Following determination of isokinetic peak torque (PT) and the isometric maximum voluntary contraction (MVC), 10 adults (mean+/-SD age=22.8+/ 2.7yrs) performed randomly ordered, submaximal step muscle actions of the dominant forearm flexors in 20% increments from 20% to 80% PT and MVC. Surface MMG signals were recorded simultaneously from a contact sensor and an accelerometer placed over the belly of the biceps brachii muscle. During the isokinetic and isometric muscle actions, the contact sensor and accelerometer resulted in linear increases in normalized MMG amplitude with torque (r(2) range=0.84-0.97) but the linear slope of the normalized MMG amplitude versus isokinetic torque relationship for the accelerometer was less (p<0.10) than that of the contact sensor. There was no significant (p>0.05) relationship for normalized MMG mean power frequency (MPF, %max) versus isokinetic and isometric torque for the contact sensor, but the accelerometer demonstrated a quadratic (R(2)=0.94) or linear (r(2)=0.83) relationship for the isokinetic and isometric muscle actions, respectively. There were also a number of significant (p<0.05) mean differences between the contact sensor and accelerometer for normalized MMG amplitude or MPF values. These findings indicated that in some cases involving dynamic and isometric muscle actions, the contact sensor and accelerometer resulted in different torque-related responses that may affect the interpretation of the motor control strategies involved. PMID- 16243543 TI - Music in dreams. AB - Music in dreams is rarely reported in scientific literature, while the presence of musical themes in dreams of famous musicians is anecdotally reported. We did a systematic investigation to evaluate whether the occurrence of musical dreams could be related to musical competence and practice, and to explore specific features of dreamt pieces. Thirty-five professional musicians and thirty non musicians filled out a questionnaire about the characteristics of their musical activity and a structured dream log on the awakening for 30 consecutive days. Musicians dream of music more than twice with respect to non-musicians; musical dreams frequency is related to the age of commencement of musical instruction, but not to the daily load of musical activity. Nearly half of the recalled music was non-standard, suggesting that original music can be created in dreams. PMID- 16243544 TI - Application of Brodmann's area templates for ROI selection in white matter tractography studies. AB - Brodmann's areas are part of the common vernacular used by neuroscientists to indicate specific location of brain activity in functional brain imaging studies. Here, we have employed a template based on the Brodmann's areas as a means of compartmentalizing underlying white matter pathways. White matter tractography was performed on the diffusion tensor data of sixteen subjects using a streamline tracking technique with Runge-Kutta integration. After co-registration, the Brodmann template was utilized for ROI selection. Tracts were segmented based on their termination in a particular area of the template. Binary masks were generated based on the tractography segmentation for a given Brodmann's area in each individual subject. Following registration to a normalized coordinate space, the binary masks were averaged, generating a map that estimates the probability of tractography connectivity for particular white matter pathways to a specific Brodmann's area. The probability maps were color-coded and overlaid on anatomical images to provide perspective. In this study, particular attention was given to the areas of the frontal cortex. A composite map of these areas was generated by assigning each voxel to the Brodmann's area with the highest probability of connectivity, based on the average results. The average maps generated with this method reveal consistent patterns of connectivity across subjects. The use of a normalized template for ROI selection automates the process of segmenting tractography data, making it particularly useful for multi-subject studies. In the future, this method could be used to help elucidate relationships between function and anatomical structure. PMID- 16243545 TI - Effects of oxygen saturation on BOLD and arterial spin labelling perfusion fMRI signals studied in a motor activation task. AB - Effects of oxygen availability on blood oxygenation level dependent (BOLD) and arterial spin labelling (ASL) perfusion functional magnetic resonance imaging (fMRI) signal changes upon motor activation were studied. Mild hypoxic hypoxia was induced by reducing the inspired oxygen content (FIO(2)) to 12%, decreasing blood oxygen saturation (Y) from 0.99 +/- 0.01 to 0.85 +/- 0.03. The fMRI signal characteristics were determined during finger tapping. BOLD activation volume decreased as a function of declining Y in the brain structures involved in execution of the motor task, however, the BOLD signal increase in activated parenchyma was not influenced by Y. ASL fMRI showed that the baseline CBF of 61.8 +/- 3.6 ml/100 g/min was not affected by hypoxic hypoxia. Similar to the BOLD fMRI, the volume of motor cortex areas displaying increase in perfusion by ASL fMRI decreased, but the signal change due to perfusion increase was not influenced in hypoxia. The present fMRI results show distinct patterns of haemodynamic and metabolic responses in the brain to motor task between normoxia and hypoxia. On one hand, neither BOLD nor ASL fMRI signal changes are influenced by hypoxia during motor activation. On the other hand, hypoxia attenuates increase in both BOLD and perfusion fMRI signals upon finger tapping from the levels determined in normoxia. These observations indicate that haemodynamic and metabolic responses may be heterogeneous in brain during execution of motor functions in mild hypoxia. PMID- 16243547 TI - The effect of a passive muscle stretching protocol on the articular cartilage. AB - OBJECTIVE: The aim of this study is to evaluate the articular cartilage alterations of rat ankles, after applying unilateral cyclic passive muscle stretching protocol in previously immobilized rats. METHODS: Twenty-two male albino rats divided into four groups, I--immobilized; IS--immobilized and stretched; S--stretched and C--control, were used in this experiment. The I and IS groups were immobilized for 4 weeks. In the muscle stretching protocol the treated ankle joint (groups IS and S) was manually full dorsal flexed 10 times for 60s with a 30s interval between each 60s period, 7 days a week for 3 weeks, to stretch the ankle plantar flexors muscle group. The right hind limb was free to move. At the end of the experiment, the ankles were removed, processed in paraffin and stained with hematoxylin-eosin and Safranin-O. Two blinded observers evaluated cellularity, chondrocyte cloning and Safranin-O staining through light microscopy. And a morphometric study was carried out using a hand count of chondrocyte cells and cartilage thickness measurement. RESULTS: No significant effect of solely muscle stretching concerning cellularity, chondrocyte cloning and Safranin-O staining parameters was detected. However, IS group presented a significantly higher reduction of proteoglycans content than the solely stretched and solely immobilized groups and the morphometric analysis showed significant cellularity increase without thickness alteration compared to control. CONCLUSIONS: These findings suggest that the stretching protocol used was harmful to the previously immobilized articular cartilage. However, the same stretching protocol did not harm the cartilage of non-immobilized groups. PMID- 16243546 TI - Abnormalities of hippocampal surface structure in very mild dementia of the Alzheimer type. AB - To better define the pattern of hippocampal deformity early in the course of Alzheimer's disease, we compared the pattern of hippocampal surface variation in subjects with very mild dementia of the Alzheimer type (DAT) and nondemented subjects. The surface of the hippocampus was divided a priori on a neuroanatomical template into three zones approximating the locations of underlying subfields [Csernansky, J.G., Wang, L., Swank, J., Miller, J.P., Gado, M., McKeel, D., Miller, M.I., Morris, J.C., 2005. Preclinical detection of Alzheimer's disease: hippocampal shape and volume predict dementia onset in the elderly. NeuroImage 25, 783--792]; i.e., a lateral zone (LZ) approximating the CA1 subfield, a superior zone (SZ) approximating the combined CA2, CA3, CA4 subfields and the gyrus dentatus (GD), and an inferior-medial zone (IMZ) approximating the subiculum. Large-deformation high-dimensional brain mapping (HDBM-LD) was used to generate the hippocampal surfaces of all subjects and to register the surface zones across subjects. Average variations within each zone were calculated for the subjects with very mild DAT as compared to the average of the nondemented subjects. After correcting for multiple comparisons, the very mild DAT subjects showed significant inward variation in the left and right LZ, the left and right IMZ, but not in the left and right SZ as compared to nondemented subjects. In logistic regression analyses, inward variation of the left and right LZ or IMZ by 0.1 mm relative to the average of the nondemented subjects increased the odds of the subject being a very mild DAT subject (range 1.18 to 1.57) rather than being a nondemented subject. The odds ratios for the left and right SZ were not significant. These results represent a replication of our previous findings [Csernansky, J.G., Wang, L., Joshi, S., Miller, J.P., Gado, M., Kido, D., McKeel, D., Morris, J.C., Miller, M.I., 2000. Early DAT is distinguished from aging by high-dimensional mapping of the hippocampus. Neurology 55, 1636--1643.] and suggest that inward deformities of the hippocampal surface in proximity to the CA1 subfield and subiculum can be used to distinguish subjects with very mild DAT from nondemented subjects. PMID- 16243548 TI - Isolation of Mycoplasma bovis from bovine clinical mastitis cases in Northern Greece. AB - Mycoplasma bovis was detected in 18/219 (8.2%) quarter milk samples collected from cases of bovine clinical mastitis in Northern Greece between November 1997 and March 1999. The cases occurred in 2/37 (5.4%) of the herds examined. The micro-organism was isolated from bulk milk samples (BTS) from the two positive herds but was not isolated from 111 composite milk samples collected from clinically healthy cows from all 37 herds. Isolates were identified as M. bovis by polymerase chain reaction (PCR) assay. Other micro-organisms were also isolated from the M. bovis positive samples. The M. bovis-positive cows had all been imported into Greece from other European countries. PMID- 16243549 TI - Paediatric cancer pain management using the WHO analgesic ladder--results of a prospective analysis from 2265 treatment days during a quality improvement study. AB - OBJECTIVE: To collect data on pain management in paediatric oncology with respect to the WHO ladder approach. SETTING, DESIGN, PATIENTS AND METHODS: Eight German tertiary care paediatric oncology centres prospectively documented all their in patient pain treatment courses from June 1999 to December 2000. Pain was scored using a 1-6 faces scale. RESULTS: Two hundred and twenty four patients (median age, 9 years; range 0.2-32.1) were enrolled. Three hundred and thirty three pain episodes comprising a total of 2265 treatment days were documented. Pain was mostly therapy associated. The most frequently administered non-opioid analgesics were dipyrone and paracetamol. On WHO step 2, tramadol was almost the only opioid used. During tramadol monotherapy average daily pain scores were lower than with a combination of tramadol and non-opioid analgesics. On WHO step 3, morphine was at least part of the analgesic regimen on most treatment days. Strong opioids were combined with a non-opioid analgesic on 41% of the treatment days. The mean intravenous morphine equivalence dose was 0.034 mg/kg/h. During opioid and non opioid combination therapy, adverse effects were more frequent, and average pain scored higher than on opioid monotherapy. CONCLUSIONS: WHO-guidelines were closely followed in Germany and seem to provide effective analgesia for children with cancer pain. In our patient group there is no evidence that a combination of an opioid with a non-opioid is more effective than opioid therapy alone in in patient paediatric oncology pain treatment. PMID- 16243550 TI - A DOQSY approach for the elucidation of torsion angle distributions in biopolymers: application to silk. AB - Silk from the wild silkworm Samia cynthia ricini with a molecular mass of about 300kDa consists of alternating repeats of nano-crystalline poly-(Ala) and non crystalline glycine-rich domains. The backbone torsion angles between pairs of these two amino acids is determined by DOQSY solid-state NMR spectroscopy: the alanine-rich domains are predominantly in a beta-sheet conformation, whereas the glycine-rich domains are found to be partially in an extended beta-sheet conformation and partially in an approximately 3(1)-helical conformation. In the cast film from liquid silk significantly different secondary structures were found: the alanine-rich domains are alpha-helical conformation, whereas the results for glycine-labelled sample are explained by a random-coil state. A detailed error analysis of the technique is presented. PMID- 16243551 TI - Simultaneous measurement of D and T2 using the distant dipolar field. AB - The presence of long-range dipolar fields in liquids is known to introduce a non linear term in the Bloch-Torrey equations which is responsible for many interesting effects in nuclear magnetic resonance as well as in magnetic resonance imaging. We show here, for the first time, that the diffusion coefficient D and the spin-spin relaxation time T2 can be obtained simultaneously from the time evolution profile of the long-range dipolar field refocused signal. In a COSY Revamped by Z-asymmetric Echo Detection sequence, the analytical first order approximation solution of the Bloch-Torrey equations modified to include the effect of the distant dipolar field is used to demonstrate the technique in an experiment using doped water. PMID- 16243552 TI - Integration of digital signal processing technologies with pulsed electron paramagnetic resonance imaging. AB - The integration of modern data acquisition and digital signal processing (DSP) technologies with Fourier transform electron paramagnetic resonance (FT-EPR) imaging at radiofrequencies (RF) is described. The FT-EPR system operates at a Larmor frequency (L(f)) of 300MHz to facilitate in vivo studies. This relatively low frequency L(f), in conjunction with our approximately 10MHz signal bandwidth, enables the use of direct free induction decay time-locked subsampling (TLSS). This particular technique provides advantages by eliminating the traditional analog intermediate frequency downconversion stage along with the corresponding noise sources. TLSS also results in manageable sample rates that facilitate the design of DSP-based data acquisition and image processing platforms. More specifically, we utilize a high-speed field programmable gate array (FPGA) and a DSP processor to perform advanced real-time signal and image processing. The migration to a DSP-based configuration offers the benefits of improved EPR system performance, as well as increased adaptability to various EPR system configurations (i.e., software configurable systems instead of hardware reconfigurations). The required modifications to the FT-EPR system design are described, with focus on the addition of DSP technologies including the application-specific hardware, software, and firmware developed for the FPGA and DSP processor. The first results of using real-time DSP technologies in conjunction with direct detection bandpass sampling to implement EPR imaging at RF frequencies are presented. PMID- 16243553 TI - A novel liquid crystalline system for partial alignment of polar organic molecules. AB - A new system for partial alignment of polar organic molecules to measure residual dipolar couplings in NMR consists of a 1:1 or a 2:1 mixture of water and DMSO including 3-13% n-alkylpentaethylene glycol as the surfactant. Temperature and concentration dependence of the alignment system are investigated and, as examples, the 13C,1H residual dipolar couplings for the amino acid methionine 1 and for an alpha-methylene-gamma-butyrolactone 2 have been obtained and are compared with those obtained from the alignment media consisting of n alkylpentaethylene glycol, n-alkyl alcohol and water. PMID- 16243554 TI - Effect of pressure on the conformation of proteins. A molecular dynamics simulation of lysozyme. AB - The effect of pressure on the structure and mobility of lysozyme was studied by molecular dynamics computer simulation at 1 and 3 kbar (1 atm = 1.01325 bar = 101.325 kPa). The results have good agreement with the available experimental data, allowing the analysis of other features of the effect of pressure on the protein solution. The studies of mobility show that although the general mobility is restricted under pressure this is not true for some particular residues. From the analysis of secondary structure along the trajectories it is observed that the conformation under pressure is more stable, suggesting that pressure acts as a 'conformer selector' on the protein. The difference in solvent-accessed surface (SAS) with pressure shows a clear inversion of the hydrophilic/hydrophobic SAS ratio, which consequently shows that the hydrophobic interaction is considerably weaker under high hydrostatic pressure conditions. PMID- 16243555 TI - Homology modeling of the DNA-binding domain of human Smad5: a molecular model for inhibitor design. AB - Members of the Smad protein family function as signal transducers of the transforming growth factor (TGF-beta) superfamily proteins. The human Smad5 protein, a signal transducer downstream of TGF-beta/BMP receptors, is composed of N-terminal DNA binding domain (MH1) and C-terminal protein-protein interaction domain (MH2) connected together by a linker motif. We used homology-modeling techniques to generate a reliable molecular model of the Smad5 MH1 domain based on the crystal structure of Smad3 MH1 domain. Our study presents the structural features of a BMP-regulated, R-Smad subfamily member (consisting of Smad1, Smad5 and Smad8) for the first time. This model provides a structural basis for explaining both functional similarities and differences between Smad3 and Smad5. Also, the structural model of this molecular target would be useful for structure based inhibitor design because of its high accuracy. The results of our study provide important insights into understanding the structure-function relationship of the members of the Smad protein family and can serve to guide future genetic and biochemical experiments in this area. PMID- 16243556 TI - A reactivity index study to rationalize the effect of dopants on Bronsted and Lewis acidity occurring in MeAlPOs. AB - The influence of both bivalent and trivalent metal substituents from a range of metal cation (Co, Mn, Mg, Fe and Cr) on the acidic property (both Bronsted and Lewis) of metal substituted aluminum phosphate MeAlPOs is monitored. The influence of the environment of the acid site is studied both by localized cluster and periodic calculations to propose that the acidity of AlPOs can be predictable with accuracy so that AlPO material with desired acidity can be designed. A semi-quantitative reactivity scale within the domain of hard soft acid-base (HSAB) principle is proposed in terms of the metal substitutions using density functional theory (DFT). It is observed that for the bivalent metal cations Lewis acidity linearly increases with ionic size, where as the Bronsted acidity is solely dependent on the nearest oxygen environment. Intramolecular and intermolecular interactions show that once active site of the interacting species is identified, the influence of the environment can be prescribed. Mg(II)-doped AlPO-34 shows highest Bronsted acidity and whereas Cr(III)-doped species shows lowest acidity. Fe(II)/Fe(III)-doped AlPO-34 shows highest Lewis acidity, whereas Mn(III), Mg (II) shows lowest acidity. PMID- 16243558 TI - Hemophagocytic syndrome associated with visceral leishmaniasis. PMID- 16243557 TI - Association of estrogen receptor alpha gene polymorphisms with bone mineral density in postmenopausal Indian women. AB - Bone mineral density (BMD) is the major determinant of osteoporotic fracture risk with a particular genetic background. However, consensus on the association of BMD with specific gene locus has not been reached. In the present study, we investigated the potential association of estrogen receptor alpha (ER alpha) gene intron I polymorphisms with BMD in 246 postmenopausal Indian women (average age 54.2+/-3.4 years). All the subjects were genotyped for XbaI and PvuII polymorphisms and underwent BMD measurements at spine and hip by dual energy X ray absorptiometery. The average BMD of subjects with the genotypes XX and PP (absence of restriction sites for XbaI and PvuII, respectively) was 12.7 and 5.4% higher at the spine and 13.1 and 4.6% higher at the hip, respectively, than those with genotypes xx and pp. In age vs. BMD scatterplot, the intercept and slope of regression lines for genotypes xx and pp at spine and hip demonstrated comparatively rapid decrease in BMD across the age. The genotype XX was significantly prevalent (p<0.001) in women with normal bone mass (32%) and genotype xx in women with osteoporotic bone mass (35.3%), within the group. A significantly higher relative risk was associated with xx genotype. The study concludes that genetic variations at ER alpha gene locus, perhaps, are associated with BMD in Indian women and may influence some determinant of bone metabolism resulting in accelerated bone loss with age. PMID- 16243559 TI - Diagnosing pneumonia in rural Thailand: Digital cameras versus film digitizers for chest radiograph teleradiology. AB - BACKGROUND: Accurate surveillance for pneumonia requires standardized classification of chest radiographs. Digital imaging permits rapid electronic transfer of data to radiologists, and recent improvements in digital camera technology present high quality, yet cheaper, options. METHODS: We evaluated the comparative utility of digital camera versus film digitizer in capturing chest radiographs in a pneumonia surveillance system in rural Thailand using a panel of radiologists; the gold standard was the hard-copy radiograph. We calculated sensitivity and specificity and conducted a receiver operator characteristics (ROC) analysis. RESULTS: Of the 192 radiographs from patients with clinical pneumonia, 166 (86%) were classified as pneumonia on the hard copies. Sensitivity and specificity for identifying pneumonia were 89% and 73% for the camera and 90% and 65% for the digitizer. In the ROC analysis, there was no statistically significant difference in the area under the curve (camera, 0.86; film digitizer, 0.91, p = 0.29). The digital camera set cost 965 dollars compared to 3000 dollars for the film digitizer. CONCLUSION: Detection of pneumonia was not measurably compromised by using digital cameras compared with film digitizers. The 3-fold lower cost of the digital camera makes this technology an affordable and widely accessible alternative for surveillance systems, vaccine trials, and perhaps clinical use. PMID- 16243560 TI - [Combined flow cytometry determination of S-phase fraction and DNA ploidy is an independent prognostic factor in node-negative invasive breast carcinoma: review of a series of 271 patients with stage I and II breast cancer]. AB - PURPOSE: To assess the significance of S-phase fraction (SPF) and DNA ploidy evaluated by DNA flow cytometry as prognostic markers in stage I or II breast cancer. PATIENTS AND METHODS: A series of 271 patients, treated by surgery, radiotherapy+/-systemic therapy was analysed (median follow up: 64 months). Standardized flow cytometry cell preparation from frozen samples and consensus rules for data interpretation were followed. Three SPF classes were defined on the basis of tertiles after adjustment for ploidy. Four groups were defined based on combinations of DNA ploidy (DIP: diploid; ANEUP: aneuploid) and SPF: DIP and low SPF (DL, N=37), DIP and medium or high SPF (DMH, N=76), ANEUP and low SPF (AL, N=24), ANEUP and medium or high SPF (AMH, N=68). Local control rate (LCR), disease-free survival (DFS), metastasis-free survival (MFS), and overall survival (OS) were correlated with DNA ploidy, SPF, DL to AMH groups, T and N stages, SBR grading, age, and hormonal status on univariate and multivariate analysis (Cox model). RESULTS: On univariate analysis, DFS and LCR were higher for DIP tumours. High SPF values were associated with shorter DFS. LCR, MFS, DFS, and OS rates were significantly different with an increasingly poorer prognosis from DL to AMH. On multivariate analysis, groups DL to AMH, histological node involvement and T stage were independently associated with MFS, and DFS. In N- patients, DL to AMH remained independent for MFS and DFS. For SBR III tumours, MFS and OS were significantly different in DL to AMH groups. These results strongly support the use of combined evaluation of DNA ploidy and SPF as independent parameters in clinical trials for N- stage I and II breast cancer. PMID- 16243561 TI - Vaccines based on the native HIV Tat protein and on the combination of Tat and the structural HIV protein variant DeltaV2 Env. AB - The promising results obtained with the HIV-1 Tat-based vaccines in mice, monkeys and humans, a better understanding of Tat immunomodulatory functions, as well as evidence that vaccination with trimeric V2 loop-deleted HIV-1 Env induces cross clade neutralizing antibodies led to the rational design of a novel vaccine based on the combination of Tat and V2-deleted Env. PMID- 16243562 TI - Role of Rac1 in Escherichia coli K1 invasion of human brain microvascular endothelial cells. AB - Escherichia coli K1 invasion of human brain microvascular endothelial cells (HBMEC) requires the reorganization of host cytoskeleton at the sites of bacterial entry. Both actin and myosin constitute the cytoskeletal architecture. We have previously shown that myosin light chain (MLC) phosphorylation by MLC kinase is regulated during E. coli invasion by an upstream kinase, p21-activated kinase 1 (PAK1), which is an effector protein of Rac and Cdc42 GTPases, but not of RhoA. Here, we report that the binding of only Rac1 to PAK1 decreases in HBMEC upon infection with E. coli K1, which resulted in increased phosphorylation of MLC. Overexpression of a constitutively active (cAc) form of Rac1 in HBMEC blocked the E. coli invasion significantly, whereas overexpression of a dominant negative form had no effect. Increased PAK1 phosphorylation was observed in HBMEC expressing cAc-Rac1 with a concomitant reduction in the phosphorylation of MLC. Immunocytochemistry studies demonstrated that the inhibition of E. coli invasion into cAc-Rac1/HBMEC is due to lack of phospho-MLC recruitment to the sites of E. coli entry. Taken together the data suggest that E. coli modulates the binding of Rac1, but not Cdc42, to PAK1 during the invasion of HBMEC. PMID- 16243563 TI - Mucosal immune responses are related to reduction of bacterial colonization in the stomach after therapeutic Helicobacter pylori immunization in mice. AB - The aim of this study was to investigate the capacity of oral and parenteral therapeutic immunization to reduce the bacterial colonization in the stomach after experimental Helicobacter pylori infection, and to evaluate whether any specific immune responses are related to such reduction. C57BL/6 mice were infected with H. pylori and thereafter immunized with H. pylori lysate either orally together with cholera toxin or intraperitoneally (i.p.) together with alum using immunization protocols that previously have provided prophylactic protection. The effect of the immunizations on H. pylori infection was determined by quantitative culture of H. pylori from the mouse stomach. Mucosal and systemic antibody responses were analyzed by ELISA in saponin extracted gastric tissue and serum, respectively, and mucosal CD4+ T cell responses by an antigen specific proliferation assay. Supernatants from the proliferating CD4+ T cells were analyzed for Th1 and Th2 cytokines. The oral, but not the parenteral therapeutic immunization induced significant decrease in H. pylori colonization compared to control infected mice. The oral immunization resulted in markedly elevated levels of serum IgG+M as well as gastric IgA antibodies against H. pylori antigen and also increased H. pylori specific mucosal CD4+ T cell proliferation with a Th1 cytokine profile. Although the parenteral immunization induced dramatic increases in H. pylori specific serum antibody titers, no increases in mucosal antibody or cellular immune responses were observed after the i.p. immunization compared to control infected mice. These findings suggest that H. pylori specific mucosal immune responses with a Th1 profile may provide therapeutic protection against H. pylori. PMID- 16243565 TI - [Recording expulsive forces during childbirth using intercostal muscle electromyogram: a pilot study. Gynecol Obstet Fertil 2005;33:299-303]. PMID- 16243564 TI - Vaccines based on Nef and on Nef/DeltaV2 Env. AB - Modified vaccinia virus Ankara (MVA) is a potent vaccine vector, which proved its safety, immunogenity and efficacy in preclinical and clinical studies. The rational for the development of a vaccine against HIV based on the regulatory protein Nef delivered by MVA combined with a V2-deleted Env protein is discussed. PMID- 16243566 TI - [Female condom or male condom: offer a choice!]. AB - OBJECTIVES: As in the entire Ile-de-France, the Val-de-Marne is facing an increase in female contamination by HIV, a clear increase in STI and furthermore the number of legal abortions is still high. Various recent studies have emphasized the decrease in condom use since 1998. Can the possibility of proposing another condom, more specifically controlled by women, boost condom use? PATIENTS AND METHODS: The conseil general (local government authority) thus initiated a study in the general population, from January 1st 2001 to December 31st 2001, in order to study the criteria of acceptability of the female condom (FC). RESULTS: Although the women coming to consultation had no specific requests a quarter finally tested the female condom during sexual intercourse. Among them 4 out of 10 are ready to use it again. Curiosity was the primary motivation of those who accepted to test the female condom (77 %). Women who were more at ease with their sexuality were twice more likely to test the female condom. Three main advantages were put forward by the women who tested the FC: the strength of the FC, the fact that insertion was possible prior to the sexual act and the possibility of postponing removal, the latter being considered a supplementary advantage. The disadvantages most frequently cited before use were its appearance and difficult insertion process. The frequency of utilisation of the FC was multiplied by three when women practised putting the condom in place in a non sexual context. However, even when they recognized one or several disadvantages women accepted to test it. In addition, whenever a woman finds at least one advantage before use she is twice more likely to test it. This is where counselling by professionals in charge of presenting the FC becomes a deciding factor. DISCUSSION AND CONCLUSIONS: The chance of using at least one sort of condom is increased when a choice of condoms is available. When both male and female condoms are available, the responsibility for prevention is better balanced within the couple: each partner can then be in charge of his or her own health. However the adoption of a barrier method such as the condom, means that the individual is conscious of such a need and finds that there are benefits mainly staying in good health. However the insufficient number of prevention and health promotion campaigns in France does not favour such an awareness. The availability of inexpensive female condoms in pharmacies, supermarkets as well as free distribution by certain institutions remains an unresolved question. PMID- 16243567 TI - [Ultrasound-guided versus clinical touch embryo transfer]. PMID- 16243568 TI - [Decreased fetal movements in the third trimester: what to do?]. AB - OBJECTIVE: To appreciate at the end of pregnancy, in a low-risk pregnant population, the interest of a screening for fetal well-being in case of decreased fetal movements. To define the most adapted screening. PATIENTS AND METHODS: Retrospective study over a complete year of the patients having consulted in the same center for decreased fetal movements and subjected to the same screening for fetal activity in hospitalization during 48 hours. This screening included a study of fetal heart rate repeated three times a day, a fetal biophysical profile scoring, an umbilical artery Doppler, a Kleihauer-Betke testing, and an amnioscopy. RESULTS: One hundred and sixty patients were identified, representing 6.1% of pregnancies followed in the center. There was no relation between the age, the parity of the patients and the probability to consult for a decrease of fetal movements. Nevertheless the antecedents of pathological pregnancy or fetal malformation were frequent. Twenty-one percent of the deliveries were induced for a global rate of 18% in the center. Twenty-eight percent of the patients had a cesarean section for a global rate of 22.8%. Five percent of fetuses were at risk for prenatal asphyxia on the data of the screening. Fetal heart rate was abnormal in 3.75% of cases, fetal biophysical profile score pathological in 3.1% of cases. Just one umbilical Doppler was highly pathological. No meconium amniotic fluid was found. Two Kleihauer-Betke tests were disturbing. At the time of delivery, 28% of fetuses presented a funicular abnormality, 4.3% a severe growth restriction, 4.3% a malformation. One child only had an anemia. There was no perinatal mortality. DISCUSSION AND CONCLUSION: Screening for fetal vitality remains necessary in case of decreased fetal movements. It has to associate the study of fetal heart rate and the fetal biophysical profile with a Kleihauer Betke testing. In a low-risk pregnant population, the study of fetal Doppler velocimetry is not profitable. Amnioscopy presents not enough interest. It is necessary to insist with the patients on the necessity of consulting in case of decreased fetal movements even in the approach of the term. PMID- 16243569 TI - [Cavernous hemangioma of the breast: a case report]. AB - Cavernous hemangiomas of the breast are uncommon. Clinical diagnosis is rather difficult. Generally there are coincidental microscopic findings. We present a case of a 67-year-old woman which commenced as a 6 cm palpable mass of the right breast. The use of mammography, ultrasound and MRI facilitate diagnosis. Large surgical excision of the lesion was performed and histology allowed to find a cavernous hemangioma without cellular atypia. PMID- 16243570 TI - [Necrotizing fasciitis of the perineum secondary to a surgical treatment of Bartholin's gland abscess]. AB - Necrotizing fasciitis of the perineum is a rare but of fast evolution, and potentially fatal infectious disease process. It is characterized by progressive inflammation and extensive necrosis of subcutaneous tissue involving the fascia and other adjacent tissues. This infection may be idiopathic or secondary to local trauma or pelvic surgery. Its mortality rate is 20%. We report a case of necrotzing fasciitis of the perineum in a 34-year-old woman following incision and drainage of Bartholin's gland abscess. Streptococcus A, Proteus mirabilis, Escherichia coli, and Candida albicans were isolated. Intravenous broad spectrum antibiotic therapy was promptly instituted. Concurrent surgical debridement of all necrotic areas was required. Post debridement therapy required a long period of dressing changes until cicatrisation. Necrotizing fasciitis of the perineum is a surgical emergency. Early diagnosis and prompt aggressive debridement are the keys to successful management. PMID- 16243571 TI - [Assessment of human papilloma virus (HPV) testing in primary screening for cervical cancer in France (May 2004). Gynecol Obstet Fertil 2005;33:357-360]. PMID- 16243572 TI - [Recording expulsive forces during childbirth using intercostal muscle electromyogram: a pilot study. Gynecol Obstet Fertil 2005;33:299-303]. PMID- 16243573 TI - [HRT: abandon? Menopausal hormone replacement: future]. PMID- 16243574 TI - [Microbiological exams and screenings during pregnancy]. AB - Infections due to bacteria, virus or parasites are frequent during pregnancy and can have severe consequences. Moreover, the vaginal carriage of commensal bacteria or dramatic changes in the equilibrium of the vaginal flora can induce complications and cause fetal contaminations. With the help of the guidelines set up by the French Agency of Accreditation and Evaluation in Health (Anaes), the authors detail the microbiological exams and screening which take part in the management of different situations at risk. PMID- 16243575 TI - [Optimization of artificial inseminations with donor semen: a four-year experience]. AB - OBJECTIVE: Our aim was to analyse the results of a donor insemination program using ovarian stimulation, swim-up sperm preparation and intrauterine insemination proposed to women with a maximum age of 39. Incidence of several clinical and biological parameters on success rates was investigated. PATIENTS AND METHODS: Retrospective analysis of the results of 249 cycles performed in 106 couples during a four-year period is reported. RESULTS: Overall pregnancy rate of 28.1% and delivery rate of 22% per cycle were achieved, with a multiple pregnancy rate of 11.4%. Most of the pregnancies (84%) were obtained before the fourth insemination. Among the different parameters studied the total number of motile sperm inseminated was found to be the most important factor for success rate: pregnancy rate per cycle reached 40.4% if more than 1.5 million progressive sperm were inseminated vs. 24.7% if they were less than 1.5 million (P<0.05). DISCUSSION AND CONCLUSION: In precise conditions, outcome of inseminations with donor semen can reach satisfying pregnancy rates, being a valuable help for couples suffering of long-time infertility. PMID- 16243576 TI - [Ovarian endometrioma and infertility: when not to treat? Gynecol Obstet Fertil 2005;33:416-422]. PMID- 16243577 TI - [About the recommendations of the HAS for health professionals concerning information given to pregnant women (April 2005)]. PMID- 16243578 TI - [Pregnancy in type 1 diabetes: insulin pump versus intensified conventional therapy. Gynecol Obstet Fertil 2005;33:389-394]. PMID- 16243579 TI - [Practicability of the vaginal prolapse surgery in the elderly woman]. AB - OBJECTIVE: To evaluate the vaginal prolapse cure in the elderly woman. PATIENTS AND METHODS: A retrospective study is carried out on 38 patients of more than 80 years, operated for a vaginal genital prolapse cure between November 1997 and October 2004. The patients' general state of health, the type of analgesia, the operative type and time, the per- and postoperative complications and the evolution of the patients one month after the intervention, had been assessed. RESULTS: The average age of the patients was 84.2 (80-95). The patients' general state of health, according to the ASA classification was about 55.3% ASA II, and about 44.7% ASA III. The anaesthesia was general in 92.1% and spinal in 7.9%. The operating type was vaginal hysterectomy associated to a Marion-Kelly and a posterior perineorrhaphy in 68.4%; and a simple vaginal hysterectomy, associated more or less to the installation of a TVT in 13.1%; an intervention of Lefort, a case of Richter and a case of simple colpectomy in 10.5%. The average operative time is about 43 minutes. The peroperative complications can be summed up in a conversion in laparotomy due to haemorrhage, and a case of ligation of the right urethra. The transfer in an intensive care unit during 24 hours is necessary only for one case in 38. The postoperative complications are in one case death at eleventh day as a result of a pulmonary embolism, and a case of temporospatial disorientation and a prolapse relapse at 6 months. The duration of stay in hospital is about 7 days (2-18). The surgery did not affect the autonomy of the patients at one month. DISCUSSION AND CONCLUSION: Vaginal prolapse concerning the elderly woman is workable in quite common practice; it involves a good cooperation between anaesthetist and surgeon. The vital risk is really present but relative. This functional surgery must not be delayed by the installation of a pessary which must be used only for the inoperable patients. PMID- 16243580 TI - Scales of environmental justice: combining GIS and spatial analysis for air toxics in West Oakland, California. AB - This paper examines the spatial point pattern of industrial toxic substances and the associated environmental justice implications in the San Francisco Bay Area, California, USA. Using a spatial analysis method called Ripley's K we assess environmental justice across multiple spatial scales, and we verify and quantify the West Oakland neighborhood as an environmental justice site as designated by the US Environmental Protection Agency. Further, we integrate the ISCST3 air dispersion model with Geographic Information Systems (GIS) to identify the number of people potentially affected by a particular facility, and engage the problem of non-point sources of diesel emissions with an analysis of the street network. PMID- 16243581 TI - Outward migration of Gnathostoma spinigerum in interferon alpha treated hepatitis C patient. AB - After the first dose injection of pegylated interferon alpha-2b (Peg-IFN alpha 2b) to a HCV infected Thai woman, she developed cyclic painful swelling nodules on right upper quadrant of abdomen and right anterior lower chest wall. The nodules subsided spontaneously within 1-2 days but were recurrent after every Peg IFN alpha-2b injection. She also experienced acute urticaria. After nine months of therapy, an immature male of G. spinigerum migrated out from the skin nodule shortly after a Peg-IFN alpha-2b injection as scheduled. The worm showed a head bulb bearing 8 transverse rows of spines which indicated immature stage. It had well defined four pairs of caudal papillae on posterior body part which were used to identify male gender. Painful migratory swelling and urticaria disappeared after the parasite was removed. She was continually treated and had sustained both virological and biochemical responses to HCV treatment. This case demonstrates that the outward migration of G. spinigerum may be stimulated by the injection of Peg-IFN alpha-2b. PMID- 16243583 TI - Homing to suppress: address codes for Treg migration. AB - Compelling evidence suggests that diverse types of immune reactions can be suppressed by CD25+ CD4+ regulatory T cells (Tregs). Although increasing knowledge has accumulated concerning the generation and functional properties of Tregs, relatively little attention has been paid to another key question: where does immune regulation by Tregs take place in vivo? Tregs can inhibit both the priming and the effector phase of an immune response, so suppression might occur both within lymphoid tissues and at peripheral sites during immune reactions. This leads to the hypothesis that appropriate localization is indispensable for in vivo Treg function and that the migratory behavior of Treg subsets influences their in vivo suppressive capacity. Current data suggest a division of labor between subpopulations of Tregs, which is mainly based on specialized homing patterns. PMID- 16243582 TI - Putative Diphyllobothrium nihonkaiense acquired from a Pacific salmon (Oncorhynchus keta) eaten in France; genomic identification and case report. AB - We report here a likely case of Diphyllobothrium nihonkaiense contracted in France through the consumption of a Pacific salmon imported from Canada. The species diagnosis was made by molecular analysis of two mitochondrial genes (COI & ND3). This case is rather unusual in that D. nihonkaiense has never been reported along the Pacific coast of North America. PMID- 16243584 TI - Insights into mechanisms of induction and ligands recognition in the transcriptional repressor EthR from Mycobacterium tuberculosis. AB - Mycobacterium tuberculosis EthR is a repressor of ethA, a gene encoding a mono oxygenase required for the activation of the prodrug ethionamide. Two EthR crystal structures have been reported recently, either in a ligand-bound (Frenois F, Engohang-Ndong J, Locht C, Baulard AR, Villeret V. Mol Cell 2004; 16: 301-7) or in a presumed apo conformation (Dover LG, Corsino PE, Daniels IR, Cocklin SL, Tatituri V, Besra GS, Futterer K. J Mol Biol 2004; 340: 1095-105). In order to infer the EthR induction mechanism, we have compared these structures. It appears that the two structures are in a conformation incompatible with repressor function, due to the presence in both proteins of fortuitous and structurally unrelated ligands. This observation paves the way to the design of specific drugs that could increase the sensitivity of M. tuberculosis to ethionamide. PMID- 16243585 TI - Home blood transfusion, a four-year experience. PMID- 16243586 TI - Role of oxidative stress in angiotensin-II mediated contraction of human conduit arteries in patients with cardiovascular disease. AB - BACKGROUND: Angiotensin II is a powerful vasoconstrictor involved in the development of high blood pressure and in the regulation of cardiovascular growth. Recent reports have suggested that in addition to the classical pathways involved in transducing responses to receptor activation, formation of reactive oxygen species by angiotensin II may also be involved. We investigated the importance of oxidative stress in angiotensin II induced contraction in human conduit arteries from patients with cardiovascular disease. METHODS AND RESULTS: Isometric contraction studies using human radial arteries entailed probes modulating the redox-dependent reactions to define the oxidative pathways involved in angiotensin II contraction. In situ oxidative fluorescence was employed to detect immediate superoxide tissue production in radial and internal mammary arteries. Treatment with TEMPOL, human superoxide dismutase, diphenyleneiodonium, oxypurinol, NG-monomethyl L-arginine considerably decreased contractile response to angiotensin II in radial arteries. Similarly, angiotensin II-stimulated arterial superoxide production was reduced in the presence of the above inhibitors. On the contrary, used as controls, norepinephrine vasoconstriction was not associated with increase of superoxide and neither ciprofloxacin nor aminophylline altered basal or angiotensin II induced superoxide generation. CONCLUSIONS: Our findings provide evidence for the role of oxidative pathways in contractile response of human conduit arteries to angiotensin II. Angiotensin II induced superoxide anion production may be mediated by multiple inter-dependent rate-limiting enzymes in both types of artery. Our studies may have important implication for future therapeutic approaches involving inhibition of angiotensin II mediated superoxide generation in hypertension and prevention of cardiovascular disease. CONDENSED ABSTRACT: We studied the role of oxidant species in contraction responses to angiotensin II in human conduit arteries. Treating radial artery segments with the anti-oxidants with a range of inhibitors, affecting the redox dependent pathways, markedly reduced contraction to angiotensin II. In parallel experiments, oxidative fluorescence was assessed and compared in human radial and internal mammary artery. Angiotensin II induced superoxide anion production may be mediated by multiple inter-dependent rate-limiting enzymes in both types of artery. PMID- 16243587 TI - Serum AGE-elastin derived peptides among diabetic children. AB - The purpose of the study was to measure advanced glycated end products (AGE) of elastin in human serum. In the present study, we adapted an ELISA technique for the determination of AGE-elastin-derived peptides (AGE-EDP) in human sera of healthy and diabetic subjects. This test makes use of human aortic elastin hydrolyzed by a chemical procedure (alpha-elastin) and AGE-Hemocyanin. Polyclonal sera from rabbit against AGE-Hemocyanin and from sheep against alpha-elastin were obtained and their specificity was tested via direct and competitive ELISA. Sera of 60 Type 1 (insulin-dependent) diabetic children and 28 healthy subjects were tested. The patients with vascular complications showed significant higher levels of age, diabetes duration, systolic blood pressure (SBP), diastolic blood pressure (DBP), dose, EDP and AGE-EDP than those without vascular complications. AGE-EDP concentrations of all diabetics correlated with triglycerides (r=0.19; p=0.04). The correlation was found between AGE-EDP and DBP in the subgroup of patients with microalbuminuria+retinopathy (r=0.94; p=0.0006). The subgroup of patients with microalbuminuria (n=19) showed correlation with age (r=0.24; p=0.008), AGE-EDP (r=0.65; p=0.0001), EDP (r=0.51; p=0.0001) and SBP (r=0.33; p=0.0003). Further studies are necessary to elucidate the relationship between the serum level of AGE-elastin degradation products and diabetic vascular complications. The measurement of non-invasive markers of elastin synthesis and degradation may be useful in monitoring development and therapeutic intervention in diabetic vascular complications. PMID- 16243588 TI - Assessing the quality of RCTs on the effect of beta-elemene, one ingredient of a Chinese herb, against malignant tumors. AB - OBJECTIVE: To evaluate the quality of randomized controlled trials (RCTs) for Elemene injections, one ingredient of Chinese herb Curcuma wenyujin, for malignant tumors widely used in clinical practice in China. METHODS: We used a systematic sample of 127 reports of RCTs that used Elemene injections as an intervention. The quality of each report was assessed using the number of Consolidated Standards for Reporting of Trials (CONSORT) checklist items included, the frequency of allocation concealment and a 5-point quality assessment instrument (Jadad). RESULTS: 69.44% of the CONSORT checklist items was included in the reports. Only 2 (1.57%) RCTs reported allocation concealment by sealed envelopes. 123 (96.85%) reports described baseline demographic and clinical characteristics of each group. But only 5 (3.94%) of 127 RCTs reported statistics analysis results of baseline data. None of the reports stated in the methods section that intention-to-treat (ITT) analysis was used, although 111 (87.40%) reports described the number of participants (denominator) in each group included in each analysis. Information regarding adverse events was reported in 83.46% of the RCTs. However the quality of reports were low as assessed by the Jadad scale. CONCLUSIONS: The methodological quality of RCTs of Elemene injection against malignant tumors was low. Therefore, the effect of Elemene injection being used in clinical settings needs to be confirmed by further RCTs. Meanwhile, there is a need to supervise and urge researchers in China to conform to Good Clinical Practice (GCP) and CONSORT guidelines when reporting. PMID- 16243589 TI - Expression of xylA genes encoding xylose isomerases from Escherichia coli and Streptomyces coelicolor in the methylotrophic yeast Hansenula polymorpha. AB - The thermotolerant methylotrophic yeast Hansenula polymorpha is able to ferment xylose to ethanol at high temperatures. H. polymorpha xylose reductase and xylitol dehydrogenase are involved during the first steps of this fermentation. In this article, expression of bacterial xylA genes coding for xylose isomerases from Escherichia coli or Streptomyces coelicolor in the yeast H. polymorpha was shown. The expression was achieved by integration of the xylA genes driven by the promoter of the H. polymorpha glyceraldehyde-3-phosphate dehydrogenase gene ( HpGAP) into the H. polymorpha genome. Expression of the bacterial xylose isomerase genes restored the ability of the H. polymorpha Deltaxyl1 mutant to grow in a medium with xylose as the sole carbon source. This mutant has a deletion of the XYL1 gene encoding xylose reductase and is not able to grow in the xylose medium. The H. polymorpha Deltaxyl1(xylA) transformants displayed xylose isomerase activities, which were near 20% of that of the bacterial host strain. The transformants did not differ from the yeast wild-type strain with respect to ethanol production in xylose medium. PMID- 16243590 TI - Discovering causes and cures for cancer from gene expression analysis. AB - Tumorigenesis is governed by a series of complex genetic and epigenetic changes. Both mechanisms can result in either the silencing or aberrant expression of messages in a cell. Gene expression profiling techniques such as the serial analysis of gene expression (SAGE) or microarray analysis can provide global overviews of these changes, as well identify key genes and pathways involved in this process. This review outlines the current roles of these techniques in cancer research, and how they may contribute to finding not only mechanisms of this disease, but potential targets for therapy. PMID- 16243591 TI - Identification of the major metabolites of resveratrol in rat urine by HPLC MS/MS. AB - To identify the major metabolites of resveratrol in rat, rat urine samples were pretreated by using solid-phase extraction technique (SPE) with polyamide cartridges. And a LC-MS/MS method with electrospray ionisation (ESI), negative ion mode and collision induced dissociation (CID), was used to elucidate the structures of the major metabolites of resveratrol. According to the results of our experiment, we found that the main metabolites of resveratrol were resveratrol monoglucuronide (M1), dihydroresveratrol monosulfate (M2), resveratrol monosulfate (M3) and dihydroresveratrol (M4). PMID- 16243592 TI - The general practitioner's approach to irritable bowel syndrome: from intention to practice. AB - BACKGROUND: Although general practitioners play a critical role in the management of irritable bowel syndrome because they deal with the most patients, guidelines are developed mainly by specialists. AIMS: To evaluate the clinical features of irritable bowel patients and the general practitioners' approach to irritable bowel syndrome in Italy. SUBJECTS AND METHODS: A questionnaire focusing on the management of this syndrome was completed by 28 general practitioners. Clinical features and diagnostic and treatment measures taken in 229 patients were analysed. RESULTS: Only 35.7% of the general practitioners were familiar with the Rome II criteria. Changes in bowel habits and abdominal pain/discomfort were the most common symptoms. Constipation (74.2%) was more frequent as the main symptom than diarrhoea. Routine blood tests (76.4%) and abdominal ultrasound (42.2%) were requested more frequently than colonoscopy (31.1%). At least one specialist consultation was recommended in 63.3% of patients. Drugs (mainly antispasmodics) were prescribed more frequently for diarrhoea (91.4%) than for constipation (55.7%). CONCLUSIONS: General practitioners are little acquainted with the Rome II criteria. Diagnostic tests and specialist consultations are often recommended; antispasmodics are the most frequently prescribed drug. Guidelines should be developed together by general practitioners and gastroenterologists to effectively manage patients at a lower cost. PMID- 16243593 TI - Nell-1 induced bone formation within the distracted intermaxillary suture. AB - Maxillary bone deficiencies, such as cleft palate and underdeveloped maxilla that require bone graft or regeneration after orthopedic or surgical expansion, pose a significant biomedical burden. Nell-1 is a secreted molecule that possesses chordin-like domains and induces cranial suture bone growth and osteoblast differentiation. To accelerate bone formation in acutely distracted palatal sutures, rat organ cultures were stimulated with Nell-1 or BMP-7 for 8 days in vitro. We hypothesized that Nell-1 stimulation to the distracted palatal suture would accelerate bone formation. Distracted palates of 4-week-old male rats were maintained in an organ culture system, and tissue was either unstimulated or stimulated with Nell-1 or BMP-7 for 8 days. MicroCT was conducted to quantitate bone formation, while alcian blue staining was conducted for cartilage localization. Immunohistochemistry of Sox9 for chondrocyte proliferation, type X collagen for hypertrophic cartilage in endochondral bone formation, and bone sialoprotein for bone formation was conducted to characterize the cellular mechanism of newly developed tissues. Distracted palates cultured in the presence of Nell-1 or BMP-7 produced statistically significantly (P < 0.05) more bone and cartilage within the intermaxillary suture, relative to unstimulated control samples. While both BMP-7 and Nell-1 induced similar bone formation in the distracted suture, BMP-7 induced both chondrocyte proliferation and differentiation, while Nell-1 accelerated chondrocyte hypertrophy and endochondral bone formation. While both Nell-1 and BMP-7 are effective in forming bone in the distracted palatal suture, they are suggested to have distinctively different mechanisms. The ability of Nell-1 to accelerate bone formation within the palate suture demonstrates the versatility of Nell-1 within the craniofacial complex as well as an exciting advance in palate suture defect healing. PMID- 16243594 TI - Flipping coins in the fly retina. AB - Color vision in Drosophila melanogaster relies on the presence of two different subtypes of ommatidia: the "green" and "blue." These two classes are distributed randomly throughout the retina. The decision of a given ommatidium to take on the "green" or "blue" fate seems to be based on a stochastic mechanism. Here we compare the stochastic choice of photoreceptors in the fly retina with other known examples of random choices in both sensory and other systems. PMID- 16243595 TI - Unraveling the molecular pathways that regulate early telencephalon development. AB - The telencephalon, at the rostral end of the developing central nervous system, starts off as a sheet of neuroepithelial cells. During development, this sheet of cells becomes patterned and morphologically partitioned into areas that give rise to the adult cerebral hemispheres. How does this happen? How are telencephalic precursor cells instructed to generate myriad neural cell types in different areas and at different times as well as to change their rates of cell proliferation, differentiation, and death? The molecular pathways required for patterning the telencephalic neuroepithelium and forming the cerebral hemispheres are beginning to be unraveled. PMID- 16243596 TI - Glia-neuron interactions in nervous system function and development. AB - Nervous systems are generally composed of two cell types-neurons and glia. Early studies of neurons revealed that these cells can conduct electrical currents, immediately implying that they have roles in the relay of information throughout the nervous system. Roles for glia have, until recently, remained obscure. The importance of glia in regulating neuronal survival had been long recognized. However, this trophic support function has hampered attempts to address additional, more active functions of these cells in the nervous system. In this chapter, recent efforts to reveal some of these additional functions are described. Evidence supporting a role for glia in synaptic development and activity is presented, as well as experiments suggesting glial guidance of neuronal migration and process outgrowth. Roles for glia in influencing the electrical activity of neurons are also discussed. Finally, an exciting system is described for studying glial cells in the nematode C. elegans, in which recent studies suggest that glia are not required for neuronal viability. PMID- 16243597 TI - The novel roles of glial cells revisited: the contribution of radial glia and astrocytes to neurogenesis. AB - Astroglial cells are the most frequent cell type in the adult mammalian brain, and the number and range of their diverse functions are still increasing. One of their most striking roles is their function as adult neural stem cells and contribution to neurogenesis. This chapter discusses first the role of the ubiquitous glial cell type in the developing nervous system, the radial glial cells. Radial glial cells share several features with neuroepithelial cells, but also with astrocytes in the mature brain, which led to the name "radial glia." At the end of neurogenesis in the mammalian brain, radial glial cells disappear, and a subset of them transforms into astroglial cells. Interestingly, only some astrocytes maintain their neurogenic potential and continue to generate neurons throughout life. We discuss the current knowledge about the differences between the adult astroglial cells that remain neurogenic and act as neural stem cells and the majority of other astroglial cells that have apparently lost the capacity to generate neurons. Additionally, we review the changes in glial cells upon brain lesion, their dedifferentiation and recapitulation of radial glial properties, and the conditions under which reactive glia may reinitiate some neurogenic potential. Given that the astroglial cells are not only the most frequent cell type in an adult mammalian brain, but also the key cell type in the wound reaction of the brain to injury, it is essential to further understand their heterogeneity and molecular specification, with the final aim of using this unique source for neuronal replacement. Therefore, one of the key advances in the field of neurobiology is the discovery that astroglial cells can generate neurons not only during development, but also throughout adult life and potentially even after brain lesion. PMID- 16243598 TI - Classical embryological studies and modern genetic analysis of midbrain and cerebellum development. AB - The brain is a remarkably complex anatomical structure that contains a diverse array of subdivisions, cell types, and synaptic connections. It is equally extraordinary in its physiological properties, as it constantly evaluates and integrates external stimuli as well as controls a complicated internal environment. The brain can be divided into three primary broad regions: the forebrain, midbrain (Mb), and hindbrain (Hb), each of which contain further subdivisions. The regions considered in this chapter are the Mb and most-anterior Hb (Mb/aHb), which are derived from the mesencephalon (mes) and rhombomere 1 (r1), respectively. The dorsal Mb consists of the laminated superior colliculus and the globular inferior colliculus (Fig. 1A and B), which modulate visual and auditory stimuli, respectively. The dorsal component of the aHb is the highly foliated cerebellum (Cb), which is primarily attributed to controlling motor skills (Fig. 1A and B). In contrast, the ventral Mb/aHb (Fig. 1B) consists of distinct clusters of neurons that together comprise a network of nuclei and projections-notably, the Mb dopaminergic and Hb serotonergic and Mb/aHb cholinergic neurons (Fig. 1G and H), which modulate a collection of behaviors, including movement, arousal, feeding, wakefulness, and emotion. Historically, the dorsal Mb and Cb have been studied using the chick as a model system because of the ease of performing both cell labeling and tissue transplants in the embryo in ovo; currently DNA electroporation techniques are also used. More recently the mouse has emerged as a powerful genetic system with numerous advantages to study events underpinning Mb/aHb development. There is a diverse array of spontaneous mutants with both Mb- and Cb-related phenotypes. In addition, numerous gene functions have been enumerated in mouse, gene expression is similar across vertebrates, and powerful genetic tools have been developed. Finally, additional insight into Mb/aHb function has been gained from studies of genetic diseases, such as Parkinson's disease, schizophrenia, cancer, and Dandy Walker syndrome, that afflict the Mb/aHb in humans and have genetic counterparts in mouse. Accordingly, this chapter discusses a spectrum of experiments, including classic embryology, in vitro assays, sophisticated genetic methods, and human diseases. We begin with an overview of Mb and aHb anatomy and physiology and mes/r1 gene expression patterns. We then provide a summary of fate-mapping studies that collectively demonstrate the complex cell behaviors that occur while the Mb and aHb primordia are established during embryogenesis and discuss the integration of both anterior-posterior (A-P) and dorsal-ventral (D-V) patterning. Finally, we describe some aspects of postnatal development and some of the insights gained from human diseases. PMID- 16243599 TI - Brain development and susceptibility to damage; ion levels and movements. AB - Responses of immature brains to physiological and pathological stimuli often differ from those in the adult. Because CNS function critically depends on ion movements, this chapter evaluates ion levels and gradients during ontogeny and their alterations in response to adverse conditions. Total brain Na(+) and Cl(-) content decreases during development, but K(+) content rises, reflecting shrinkage of the extracellular and increase in the intracellular water spaces and a reduction in total brain water volume. Unexpectedly, [K(+)](i) seems to fall during the first postnatal week, which should reduce [K(+)](i)/ [K(+)](e) and result in a lower V(m), consistent with experimental observations. Neuronal [Cl( )](i) is high during early postnatal development, hence the opening of Cl(-) conduction pathways may lead to plasma membrane depolarization. Equivalent loss of K(+)(i) into a relatively large extracellular space leads to a smaller increase in [K(+)](e) in immature animals, while the larger reservoir of Ca(2+)(e) may result in a greater [Ca(2+)](i) rise. In vivo and in vitro studies show that compared with adult, developing brains are more resistant to hypoxic/ischemic ion leakage: increases in [K(+)](e) and decreases in [Ca(2+)](e) are slower and smaller, consistent with the known low level of energy utilization and better maintenance of [ATP]. Severe hypoxia/ischemia may, however, lead to large Ca(2+)(i) overload. Rises in [K(+)](e) during epileptogenesis in vivo are smaller and take longer to manifest themselves in immature brains, although the rate of K(+) clearance is slower. By contrast, in vitro studies suggest the existence of a period of enhanced vulnerability sometime during the developmental period. This chapter concludes that there is a great need for more information on ion changes during ontogeny and poses the question whether the rat is the most appropriate model for investigation of mechanisms of pathological changes in human neonates. PMID- 16243600 TI - Thinking about visual behavior; learning about photoreceptor function. AB - Visual behavioral assays in Drosophila melanogaster were initially developed to explore the genetic control of behavior, but have a rich history of providing conceptual openings into diverse questions in cell and developmental biology. Here, we briefly summarize the early efforts to employ three of these behaviors: phototaxis, the UV-visible light choice, and the optomotor response. We then discuss how each of these assays has expanded our understanding of neuronal connection specificity and synaptic function. All of these studies have contributed to the development of sophisticated tools for manipulating gene expression, assessing cell fate specification, and visualizing neuronal development. With these tools in hand, the field is now poised to return to the original goal of understanding visual behavior using genetic approaches. PMID- 16243601 TI - Critical period mechanisms in developing visual cortex. AB - Binocular vision is shaped by experience during a critical period of early postnatal life. Loss of visual acuity following monocular deprivation is mediated by a shift of spiking output from the primary visual cortex. Both synaptic and network explanations have been offered for this heightened brain plasticity. Direct experimental control over its timing, duration, and closure has now been achieved through a consideration of balanced local circuit excitation-inhibition. Notably, canonical models of homosynaptic plasticity at excitatory synapses alone (LTP/LTD) fail to produce predictable manipulations of the critical period in vivo. Instead, a late functional maturation of intracortical inhibition is the driving force, with one subtype in particular standing out. Parvalbumin-positive large basket cells that innervate target cell bodies with synapses containing the alpha1-subunit of GABA(A) receptors appear to be critical. With age, these cells are preferentially enwrapped in peri-neuronal nets of extracellular matrix molecules, whose disruption by chondroitinase treatment reactivates ocular dominance plasticity in adulthood. In fact, critical period plasticity is best viewed as a continuum of local circuit computations ending in structural consolidation of inputs. Monocular deprivation induces an increase of endogenous proteolytic (tPA-plasmin) activity and consequently motility of spines followed by their pruning, then re-growth. These early morphological events faithfully reflect competition only during the critical period and lie downstream of excitatory-inhibitory balance on a timescale (of days) consistent with the physiological loss of deprived-eye responses in vivo. Ultimately, thalamic afferents retract or expand accordingly to hardwire the rapid functional changes in connectivity. Competition detected by local inhibitory circuits then implemented at an extracellular locus by proteases represents a novel, cellular understanding of the critical period mechanism. It is hoped that this paradigm shift will lead to novel therapies and training strategies for rehabilitation, recovery from injury, and lifelong learning in adulthood. PMID- 16243602 TI - brawn for brains: the role of MEF2 proteins in the developing nervous system. AB - The myocyte enhancer factor 2 (MEF2) transcription factors were originally identified, as their family name implies, on the basis of their role in muscle differentiation. Expression of the four MEF2 proteins, however, is not restricted to contractile tissue. While it has been known for more than a decade that MEF2s are abundantly expressed in neurons, their contributions to the development and function of the nervous system are only now being elucidated. Interestingly, the emerging mechanisms regulating MEF2 in neurons have significant parallels with the regulatory mechanisms in muscle, despite the quite distinct identities of these two electrically excitable tissues. The goal of this chapter is to provide an introduction to those regulatory mechanisms and their consequences for brain development. As such, we first provide an overview of MEF2 itself and its expression within the central nervous system. The second part of this chapter describes the signaling molecules that regulate MEF2 transcriptional activity and their contributions to MEF2 function. The third part of this chapter discusses the role of MEF2 proteins in the developing nervous system and compares the analogous functions of this protein family in muscle and brain. PMID- 16243603 TI - Mechanisms of axon guidance in the developing nervous system. AB - The human brain assembles an incredible network of over a billion neurons. Understanding how these connections form during development in order for the brain to function properly is a fundamental question in biology. Much of this wiring takes place during embryonic development. Neurons are generated in the ventricular zone, migrate out, and begin to differentiate. However, neurons are often born in locations some distance from the target cells with which they will ultimately form connections. To form connections, neurons project long axons tipped with a specialized sensing device called a growth cone. The growing axons interact directly with molecules within the environment through which they grow. In order to find their targets, axonal growth cones use guidance molecules that can either attract or repel them. Understanding what these guidance cues are, where they are expressed, and how the growth cone is able to transduce their signal in a directionally specific manner is essential to understanding how the functional brain is constructed. In this chapter, we review what is known about the mechanisms involved in axonal guidance. We discuss how the growth cone is able to sense and respond to its environment and how it is guided by pioneering cells and axons. As examples, we discuss current models for the development of the spinal cord, the cerebral cortex, and the visual and olfactory systems. PMID- 16243604 TI - Characterization of two APP gene promoter polymorphisms that appear to influence risk of late-onset Alzheimer's disease. AB - Alzheimer's disease (AD) is characterized by formation of plaques of amyloid beta peptide (Abeta). Autosomally-inherited or "familial" AD had been demonstrated only in connection with coding sequence mutations. We characterized DNA-protein interaction and expression influence of two polymorphisms that occur in the promoter (C<-->T at -3829 and T<-->C at -1023, +1 transcription start site) of the Abeta precursor protein (APP) gene. We report distinct functional differences in reporter expression and in DNA-protein interaction for variant sequences in both -3829 and -1023 polymorphic regions. The -3829T variant has reduced DNA protein interaction and reporter expression compared to -3829C, while -1023C has greater DNA-protein interaction and reporter expression than -1023T. Our predictions for likely transcription factors for loss of function (-3829T) are ADR1, MIG1, and PuF, and for gain of function (-1023C) are E12/E47, ITF-2, and RFX2. Characterization of the activity of a regulatory polymorphism of the APP gene points towards understanding mechanisms that likely underlie the majority of AD cases and may contribute to promoter-based drug design. PMID- 16243605 TI - Somatic mitochondrial DNA mutations in single neurons and glia. AB - Somatic mitochondrial DNA (mtDNA) point mutations reach high levels in the brain. However, the cell types that accumulate mutations and the patterns of mutations within individual cells are not known. We have quantified somatic mtDNA mutations in 28 single neurons and in 18 single glia from post-mortem human substantia nigra of six control subjects. Both neurons and glia contain mtDNA with somatic mutations. Single neurons harbor a geometric mean (95% CI) of 200.3 (152.9-262.4) somatic mtDNA point mutations per million base pairs, compared to 133.8 (97.5 184.9) for single glia (p=0.0251). If mutations detected multiple times in the same cell are counted only once, the mean mutation level per million base pairs remains elevated in single neurons (146.9; 124.0-174.2) compared to single glia (100.5; 81.5-126.5; p=0.009). Multiple distinct somatic point mutations are present in different cells from the same subject. Most of these mutations are individually present at low levels (less than 10-20% of mtDNA molecules), but with high aggregate mutation levels, particularly in neurons. These mutations may contribute to changes in brain function during normal aging and neurodegenerative disorders. PMID- 16243606 TI - Subjective memory complaints in aging are associated with elevated cortisol levels. AB - The origin and clinical significance of subjective memory complaints among middle aged and older individuals is not well understood. Associations with objective memory impairments, personality traits or mood disturbances have been reported. Elevated cortisol levels occur in aging and depression and causal links to cognitive or emotional problems have been suggested. The goal of this study was to investigate the associations between basal and feedback indices of cortisol regulation and subjective memory impairment in a sample of healthy middle aged and older subjects (mean age 61.8 years) with (n=27) and without (n=19) subjective memory complaints. Participants with memory complaints had both higher basal cortisol levels and higher cortisol levels after dexamethasone. There was a significant group by gender interaction for basal cortisol levels, where women without memory complaints showed significantly lower cortisol levels, whereas no such difference was found for the men. All effects were not due to slight differences in depression scores. Differences in personality traits or in stress susceptibility might underlie the present findings. Future studies of memory complaints should take a comprehensive approach including relevant endocrine parameters. PMID- 16243610 TI - History of sleep medicine. AB - Sleeping and dreaming always have been a fundamental part of human existence. Most early writing on these subjects was almost entirely speculation. During the twentieth century, however, scientific observation and experimentation abounded. This article emphasizes the evolution of the key concepts and research findings that characterize sleep research and sleep medicine, crucial discoveries and developments in the formative years of the field, and those principles and practices that have stood the test of time. PMID- 16243607 TI - Estrogen's effects on central and circulating immune cells vary with reproductive age. AB - Previous work from this lab has shown that estrogen attenuates inflammatory cytokine production following brain lesions in young adult female rats, but not in older, reproductive senescent females. The present study was designed to elucidate whether these effects result from estrogen's actions on brain-resident immune cells (microglia) or on circulating immune cells recruited to the brain from blood. Microglia, harvested from the olfactory bulbs of ovariectomized young adult and reproductive senescent animals, were pretreated with 17beta-estradiol and subsequently with the bacterial endotoxin LPS. LPS treatment significantly increased the pro-inflammatory cytokine IL-1beta in microglial cultures harvested from young and senescent females, but estrogen treatment had no effect on cytokine expression in either group. In young adult-derived microglia, LPS treatment also increased nitric oxide (NO), which was attenuated by estrogen, and MMP-9, which was not affected by estrogen. Reproductive senescent-derived microglia cultures had higher basal expression of NO and MMP-9 activity as compared to those from young adult microglial cultures, although LPS did not further stimulate these inflammatory markers. In blood cultures, LPS stimulated a dose-dependent increase in the inflammatory cytokine TNF-alpha expression in both young adult and reproductive senescent animals. Estrogen replacement significantly attenuated TNF-alpha induction by LPS in blood cultures derived from young adult females. Paradoxically, estrogen replacement increased LPS induced TNF-alpha expression in blood cultures derived from reproductive senescent animals as compared to age-matched controls. The age and estrogen dependent effects on circulating immune cells found in whole blood cultures closely mimic the effects of estrogen on cytokine expression in the young and senescent animals that we reported in vivo, supporting the hypothesis that the immunosuppressive actions of estrogen replacement on neural injury may result from hormone-action on circulating immune cells. PMID- 16243608 TI - REM sleep control during aging in SAM mice: a role for inducible nitric oxide synthase. AB - Evidence that nitric oxide (NO) is involved in the regulation of rapid-eye movement sleep (REMS) is supported by recent studies. During aging, NO generation encounters marked changes mainly related to the activation of the inducible NO synthase (iNOS). To investigate links existing between iNOS and REMS impairments related to aging, we examine the age-related variations occurring in: mRNA and activity of iNOS in brainstem and frontal cortex; sleep parameters under baseline and after treatment by a selective iNOS inhibitor (AMT) in Senescence Accelerated Mice (SAM). SAMR1 (control) mice are a model of aging while SAMP8 are adequate to study neurodegenerative processes. RT-PCR analysis does not reveal significant variation in iNOS mRNA expression in both strains. However, significant age related increases in iNOS activity occur in SAMR1 but such variation is not observed in SAMP8. In baseline conditions, aging induces a slight increase in slow-wave sleep (SWS) amounts in both groups and deteriorates greatly REMS architecture in SAMP8 compared to SAMR1. AMT reduces REMS amounts for 4-6h after treatment in a dose and age-dependent manner in SAMR1. Almost no changes occur in SAMP8. Data reported suggest that NO derived from iNOS contributes to trigger and maintain REMS during aging. PMID- 16243611 TI - The neurology of sleep. AB - Neurology, by virtue of its study of the brain, is the primary medical science for the elucidation of the anatomy, physiology, pathology and, ultimately, the function of sleep. There has been nothing short of a revolution in the science of sleep over the past 50 years. From the discovery of REM sleep to the identification of Hypocretin/Orexin the basic science and clinical field of sleep medicine has blossomed. This article will explore the anatomy, physiology, biochemistry and, to a limited extent, pathophysiology of the sleep/wake centers of the brain. The field of chronobiology will also be touched upon. PMID- 16243612 TI - Clinical and technologic approaches to sleep evaluation. AB - This article discusses normal human sleep. Discoveries leading to an understanding of human sleep, electroencephalographic definitions, and general characteristics of normal human sleep are presented. Actuarial data for a first laboratory night are provided. Finally, the mechanisms governing sleep and wakefulness are reviewed and a model of normal sleep mechanisms going awry is outlined as an aid for understanding abnormal sleep associated with sleep disorders. PMID- 16243613 TI - Sleep and the gastrointestinal tract. AB - In this review, an integration of GI functioning is attempted with regard to its relationship to sleep, how this interaction may lead to complaints of sleep disorders, and the pathogenesis of some GI disorders. Data are presented to support the notion that sleep-related GER is an important factor not only in the development of esophagitis but also in the respiratory complications of GER. Although sensory functioning is altered markedly during sleep with regard to most standard sensory functions (eg, auditory), there seems to be an enhancement of some visceral sensation during sleep that seems to protect the tracheobronchial tree from aspiration of gastric contents reflux during sleep. Patients who have functional bowel disorders reveal an increase in sleep complaints compared with normal volunteers. The actual mechanisms of these disturbances remain somewhat obscure and studies do not demonstrate any consistent abnormalities in sleep patterns of these patients. Some studies show that autonomic functioning during sleep, particularly REM sleep, can distinguish patients who have IBS. Thus, the continued study of sleep and GI functioning promises to create a new dimension in the understanding of the pathophysiology of a variety of GI disorders. PMID- 16243614 TI - Conditions of primary excessive daytime sleepiness. AB - Excessive daytime somnolence is a prevalent problem in medical practice and in society. It exacts a great toll in quality of life, personal and public safety, and productivity. The causes of EDS are myriad, and careful evaluation is needed to determine the cause in each case. Although much progress has been made in discovering the pathophysiology of narcolepsy, much more remains to be understood, and far less is known about other primary conditions of EDS. Several methods have been developed to assess EDS, although each of them has limitations. Treatment is available for the great majority of cases. PMID- 16243615 TI - Sleep-related breathing disorders. AB - Sleep-related breathing disorders are a heterogeneous group of conditions that may be associated with alterations in the structure of sleep, in sleep quality, and in gas exchange during sleep. Obstructive sleep apnea represents the most frequent cause of sleep-related breathing disorders, which encompass a diversity of conditions that either complicate coexisting disease or present as primary disorders. Many of these disorders have consequences during both sleep and wakefulness and may produce substantial burden of symptoms and disease in untreated individuals. PMID- 16243616 TI - Sleep-disordered breathing and cerebrovascular disease: a mechanistic approach. AB - The observations described in this article point to the existence of increased oxidative stress and systemic inflammation in sleep apnea and have paved the way for establishing sleep apnea as an independent risk factor for cardio- and cerebrovascular morbidities. The proposed course of events is summarized in Fig. 1. It is suggested that hypoxia/reoxygenation,characteristic of sleep apnea, promotes the formation of ROS, particularly during the reoxygenation period, and can be deleterious to cells and tissues. ROS, however, regulate the activation of critical transcription factors that are redox sensitive, resulting in increased expression of sets of genes that encode proteins essential to adaptation to hypoxia (via hypoxia inducible factor I [hypoxia inducible factor-la]). Yet, redox-sensitive transcription factors (NFKB and AP-1) that elicit inflammatory pathways also are activated, thereby affecting inflammatory and immune responses by promoting activation of endothelial cells, leukocytes, and platelets. These activated cells express adhesion molecules and proinflammatory cytokines that may lead to endothelial injury and dysfunction and consequently to the development of cardio- and cerebrovascular morbidities. These may be exaggerated in patients who have sleep apnea in response to the intermittent hypoxia. PMID- 16243617 TI - Non-rapid eye movement sleep parasomnias. AB - Parasomnias are unpleasant or undesirable behavioral or experiential phenomena that occur during sleep. Once believed unitary phenomena related to psychiatric disorders, it is now clear that parasomnias result from several different phenomena and usually are not related to psychiatric conditions. Parasomnias are categorized as primary (disorders of the sleep states) and secondary (disorders of other organ systems that manifest themselves during sleep). Primary sleep parasomnias can be classified according to the sleep state of origin: rapid eye movement sleep, non-rapid eye movement sleep, and miscellaneous (those not respecting sleep state). Secondary sleep parasomnias are classified by the organ system involved. PMID- 16243618 TI - Rapid eye movement sleep parasomnias. AB - The recognition of RBD has shed additional scientific light on the "bumps in the night"; expanded knowledge of states of being and state dissociation; opened up new areas of research on brain and mind dysfunction during sleep; expanded knowledge of various neurologic disorders, particularly narcolepsy and parkinsonism; and reaffirmed the vital link between basic research and clinical medicine. Moreover, the safe and effective treatment of RBD with clonazepam is especially gratifying. PMID- 16243619 TI - Sleep and epilepsy. AB - This article examines the relationship between sleep and epilepsy, an association that has been recognized since antiquity. The mechanisms whereby sleep facilitates seizures are under investigation, although the synchronizing role of thalamocortic networks seems contributory. Recognition of the variety of generalized and partial epileptic syndromes associated with sleep, familiarity with the differential diagnosis of nocturnal spells, and awareness of the role that antiepileptic drugs and sleep disorders may play in epilepsy are helpful in evaluating patients presenting with behavioral and motor disturbances of sleep. PMID- 16243620 TI - Pharmacologic and nonpharmacologic treatments of insomnia. AB - Insomnia in its chronic form is present in high numbers of patients presenting to physicians. As older women who have medical problems have the highest rates of chronic insomnia, physicians must have a high index of suspicion and be prepared to explore various etiologic factors that might be operative. Treatment should focus on setting specific goals, with patients using strategies that combine lifestyle changes, behavioral interventions, and appropriate medications. OTC agents, sedating antidepressants at low dosages (trazodone, doxepin, amitriptyline, and others), and nonhypnotic benzodiazepines are insufficiently studied to provide evidence-based support for their use to treat chronic insomnia. Particularly in the elderly, close monitoring is needed to prevent falls, accidents, and cognitive impairment from these agents. FDA-labeled hypnotic agents are efficacious, but long-term studies have not been available until the recent release of eszopiclone in the United States. Recent work encourages the use of CBT even in patients who have used sleeping pills for several years, although the success of CBT has been less encouraging when applied to chronic insomnia sufferers who have concurrent psychiatric disorders and who have taken hypnotics for years. PMID- 16243621 TI - Restless legs syndrome. AB - Restless legs syndrome (RLS) is clinically defined as an urge to move the legs with or without paresthesia, worsening of symptoms with rest and transient improvement with activity, and worsening of symptoms in the evening and night. This is often genetic but may also occur in the setting of iron deficiency, uremia, pregnancy, neuropathy, and possibly other conditions. The pathology is probably related to central nervous system iron dysregulation. Effective treatments include dopaminergics and narcotics. Recent advances in our understanding of RLS clinical presentation, epidemiology, etiology, and treatment will be discussed. PMID- 16243622 TI - Parkinson's disease and sleep. AB - Sleep disorders are common in PD and many factors can contribute to disturbed nocturnal sleep and daytime sleepiness. Factors contributing to sleep disturbance include the presence of insomnia, mood or anxiety disorders, dementia, specific sleep disorders, PD motor disorders, and the effects of PD or medications. Patients who have PD should be interrogated about sleep disturbance and daytime sleepiness and preferably, because of underestimation of the severity of sleepiness or lack of awareness, patients should be interviewed in the presence of a close friend or relative. The ability to drive, if sleepiness is present, should be assessed and appropriate recommendations made. Treatment of sleepiness involves treating any underlying sleep disturbance and may involve the use of stimulant or alerting medications in the daytime. PMID- 16243625 TI - Supportive therapy of the toxic cow. PMID- 16243623 TI - Sleep and neuromuscular disorders. AB - Nocturnal sleep-related ventilatory alterations may occur in dis-proportion to the severity of the neuromuscular disorder. Diaphragm paralysis occurring with a neuromuscular disorder is an overlooked complication. Failure to thrive, daytime tiredness, and incapacitating fatigue may be the result of a correctable sleep related abnormality, not the result of relentless progression of a neuromuscular condition. Polysomnographic evaluation is recommended for patients who have neuromuscular disorder who develop symptoms and signs of sleep-wake abnormality or nocturnal respiratory failure. Application of noninvasive positive airway ventilation and, in some cases, administration of supple-mental oxygen may improve quality of life and prolong survival of patients who have neuromuscular disorder. PMID- 16243626 TI - Blood component transfusions. PMID- 16243627 TI - Pain management in cattle. PMID- 16243628 TI - Neonatal care of high-risk cloned and transgenic calves. PMID- 16243629 TI - Abdominal emergencies in cattle. PMID- 16243630 TI - Respiratory emergencies in cattle. PMID- 16243631 TI - Bovine reproductive emergencies. PMID- 16243632 TI - Toxicologic emergencies in cattle. PMID- 16243633 TI - Factor structure, reliability, and validity of the Penn State Worry Questionnaire: differences between African-American and White-American college students. AB - This study examined differences in the factor structure of the Penn State Worry Questionnaire (PSWQ) between African-American (n=181) and White-American (n=180) college students. Results from a confirmatory factor analysis indicated that the traditional single-factor solution did not provide the best fit for the data from either ethnic group. A multiple-group factor analysis indicated that underlying structure of Factor 1 was roughly equivalent between ethnic groups. Structure of Factor 2, however, differed between groups. Specifically, item 10 loaded on different factors for each group. In support of these analyses, an exploratory factor analyses (EFA) among White-American participants indicated the presence of a two-factor model while an EFA among African-Americans indicated the presence of three factors. Despite some overlap in the overall factor structure between ethnic groups, African-Americans scored significantly lower on the PSWQ than the White-American group. Furthermore, among African-Americans level of ethnic identity was negatively related to state and trait measures of anxiety, but unrelated to measures of depression and worry. PMID- 16243634 TI - Mental health among Hispanics and Caucasians: risk and protective factors contributing to prevalence rates of psychiatric disorders. AB - The current study examined the one-year prevalence of psychiatric disorders for Hispanics and Caucasians in a large population sample (N=4559) and explored factors that contributed to group differences. Hispanic participants (predominantly Mexican Americans) were more likely than Caucasian participants to have met the criteria for a psychiatric diagnosis in the past year, had higher one-year prevalence rates of several anxiety disorders, had greater problems meeting their basic needs, and better interpersonal functioning. Hispanic participants' problems meeting basic needs partially mediated their higher prevalence of psychiatric disorders compared to Caucasian participants. Better interpersonal functioning protected Hispanic participants against depression, panic, and substance use disorders. These findings are discussed in terms of the importance of psychosocial variables for the prevalence of psychiatric disorders. PMID- 16243635 TI - Correlates for posttraumatic stress disorder in Gulf War veterans: a retrospective study of main and moderating effects. AB - With a sample of 120 Gulf War veterans, the present study investigated the main effects of childhood and lifetime trauma, combat exposure, and coping strategies on posttraumatic stress disorder (PTSD), as well as combat exposure's moderating effects on the other variables' relationships with PTSD. Logistic regression results indicated correct classification of PTSD diagnosis for 88% of the participants, with combat exposure and avoidant coping making significant contributions to this classification. Multiple regression results indicated that lifetime trauma, combat exposure, and avoidant coping were strongly related to PTSD symptoms. Multiple regression results also revealed that combat exposure moderated the strength and direction of PTSD's relationships with childhood trauma and avoidant coping. Study findings have implications for longitudinal investigation of PTSD development and preventive interventions. PMID- 16243636 TI - A Brief Measure of Worry Severity (BMWS): personality and clinical correlates of severe worriers. AB - This report describes the development of a brief and valid self-report measure to assess severe and dysfunctional worry (the Brief Measure of Worry Severity or BMWS). Using three independent subject groups (clinical and non-clinical), the measure was used to examine the differential severity of worry in depression and anxiety and to examine the clinical and personality correlates of severe worriers. Preliminary psychometric evaluation revealed that the BMWS possesses good construct and clinical discriminant validity. Subjects reporting greater worry severity tended to be more "introverted" and "obsessional," but less "agreeable" and "conscientious." Subjects with depression only, reported less problems with worrying compared to those with co-morbid anxiety disorders. However, among the anxiety disorders, severe and dysfunctional worry was not exclusively experienced by subjects with generalized anxiety disorder (GAD). This study suggests that pathological worry is not only relevant for patients with GAD, but may be an equally detrimental cognitive activity for patients with panic disorder and obsessive-compulsive disorder. PMID- 16243637 TI - Psychometric properties of the Spanish version of the Obsessive-Compulsive Inventory--revised in a non-clinical sample. AB - The psychometric properties of a Spanish version of the Obsessive-Compulsive Inventory-Revised (OCI-R) were examined in a non-clinical student sample (n=381). A confirmatory factor analysis replicated the original six-factor structure. The total and each of the subscales of the Spanish OCI-R demonstrated moderate to good internal consistency and test-retest reliability, moderate convergent validity and good divergent validity. The Spanish version of the OCI-R retains the sound psychometric properties of the original version. PMID- 16243638 TI - Exploratory factor analysis of two measures of posttraumatic stress disorder (PTSD) symptoms in a non-clinical sample of college students. AB - This study investigated the psychometric structure of two widely utilized measures of posttraumatic symptoms in a primarily Caucasian non-clinical sample. Given the prevalence of trauma exposure in non-referred samples, measurement of resulting symptoms is a critical issue. Exploratory factor analysis was utilized to assess and compare the factor structure of the Impact of Event Scale [IES; Horowitz, M., Wilner, N., & Alvarez, W. (1979). Impact of Event Scale: a measure of subjective stress. Psychosomatic Medicine, 41, 209-218] and the Mississippi Scale for Posttraumatic Stress Disorder, Civilian version [MIS-Civ; Vreven, D. L., Gudanowski, D. M., King, L. A., & King, D. W. (1995). The Civilian Version of the Mississippi PTSD Scale-a psychometric evaluation. Journal of Traumatic Stress, 8, 91-109] in a sample of college students reporting exposure to potentially traumatic events. The psychometric structure of the IES was largely consistent with the two-factor structure widely reported in the literature, while the structure of the MIS-Civ varied considerably in this sample. Notably, non clinical samples tended to report fewer social and occupational dysfunction than clinical samples predominantly utilized in PTSD research. Implications for use of these instruments in screening samples are discussed. PMID- 16243639 TI - Are there lessons to be learned from drug development that will accelerate the use of molecular imaging probes in the clinic? AB - This special issue of the journal contains contributions from participants of the third La Jolla meeting (The Magic Bullet: A Century Later). The goal of this meeting was twofold: to review approaches to validating molecular imaging agents and to review the progress in advancing the use of molecular imaging from the bench to the bedside, with a special emphasis on how molecular imaging improves patient care and management. Drug development has changed its focus over the years. The original approach depended on direct measurements in patients, whereby, in many cases, the drug was advanced to an NDA based on physiological results (e.g., lowering blood pressure) without identifying a target. Over the past decade, the focus has been on validating a target and choosing the lead compound using combinatorial chemistry and high throughput screening, often at the expense of a focus on the biology of diseases. On the other hand, molecular imaging has been target based since its beginning because of the requirements dictated by external imaging (i.e., a target-to-nontarget ratio). This article explores the possible analogies between current targeted drug development and molecular imaging-targeted probe development with the goal of better defining the path to new molecular imaging probes for the clinic. PMID- 16243640 TI - True tracers: comparing FDG with glucose and FLT with thymidine. AB - As PET metabolic imaging becomes routine in clinical practice, there is a tendency to make imaging and data analysis fast and simple, but interpretation of these pictures by visual inspection does not do justice to the power of PET technology. Tissue data and blood data can be analyzed mathematically to provide parametric images of the PET tracer's biochemistry in terms of a transport parameter and a metabolic flux. The methods for parametric imaging with (11)C tracers of glucose and thymidine have been validated, but the short half-life of this radionuclide and the rapid metabolism of these labeled substrates to [(11)C]CO(2) have led investigators to develop (18)F analogs. While (18)F substitution at critical positions in the natural substrate can block metabolism, it has other effects on the transport and metabolism of the analog tracer. The fidelity with which analog tracers mimic tracers of the authentic substrate is critically evaluated for [(18)F]-2-fluoro-2-deoxyglucose and [(18)F]-3'-fluoro-3' deoxythymidine. PMID- 16243641 TI - Mass effect of injected dose in small rodent imaging by SPECT and PET. AB - This paper discusses the effect of mass (chemical quantity) of injected dose on positron emission tomography (PET) and single-photon emission computed tomography (SPECT). Commonly, PET or SPECT imaging study uses a "no-carrier added" dose, which contains a small amount of radioactive imaging agent (in picogram to microgram). For small animal (rodent) imaging studies, specifically targeting binding sites or biological processes, the mass (chemical quantity) in the dose may significantly modify the binding, pharmacokinetics and, ultimately, the imaging outcome. Due to differences in size and other physiological factors between humans and rodents, there is a dramatic divergence of mass effect between small animal and human imaging study. In small animal imaging studies, the mass, or effective dose (ED(50)), a dose required for 50% of receptor or binding site occupancy, is usually not directly related to binding potential (B(max)/K(d)) (measured by in vitro binding assay). It is likely that dynamic interplays between specific and nonspecific binding in blood circulation, transient lung retention, kidney excretion, liver-gallbladder flow, soft tissue retention as well as metabolism could each play a significant role in determining the concentration of the tracer in the target regions. When using small animal imaging for studying drug occupancy (either by a pretreatment, coinjection or chasing dose), the mass effects on imaging outcome are important factors for consideration. PMID- 16243642 TI - Measurement of input functions in rodents: challenges and solutions. AB - INTRODUCTION: Tracer kinetic modeling used in conjunction with positron emission tomography (PET) is an excellent tool for the noninvasive quantification of physiological, biological and molecular processes and their alterations due to disease. Currently, complex multi-compartment modeling approaches are being applied in a variety of clinical studies to determine myocardial perfusion, viability and glucose utilization as well as fatty acid metabolism and oxidation in the normal and diseased heart. These kinetic models require two key measurements of tracer activity over time, tracer activity in arterial blood (input function) and its corresponding activity in the organ of interest. The alteration in the time course of tracer activity as it travels from blood to the organ of interest describes the kinetics of the tracer. To be able to implement these approaches in rodent models of disease using small-animal PET (microPET), it is imperative that the input function is measured accurately. METHODS: The blood input functions in rodent experiments were obtained by (1) direct blood sampling, (2) direct measurement of blood activity by a beta-detecting probe that counts the activity in the blood, (3) an arterial-venous bypass (A/V shunt), (4) factor analysis of dynamic structures from dynamic PET images and (5) measurement from region-of-interest (ROI) analysis of dynamic PET images. Direct blood sampling was used as the reference standard to which the results of the other techniques were compared. RESULTS: Beta probes are difficult to operate and may not provide accurate blood input functions unless they are used intravenously, which requires complicated microsurgery. A similar limitation applies to the A/V shunt. Factor analysis successfully extracts the blood input function for mice and rats. The ROI-based method is less accurate due to limited image resolution of the PET system, which results in severe partial volume effect and spillover from myocardium. CONCLUSION: The current reference standard, direct blood sampling, is more invasive and has limited temporal resolution. With current imaging technology, image-based extraction of blood input functions is possible by factor analysis, while forthcoming technological developments are likely to allow extraction of input function directly from the images. These techniques will reduce the level of complexity and invasiveness for animal experiments and are likely to be used more widely in the future. PMID- 16243643 TI - Cy5.5-DTPA-galactosyl-dextran: a fluorescent probe for in vivo measurement of receptor biochemistry. AB - The high sensitivity of fluorescent reporters offers an opportunity to analytically probe the biochemistry of in vivo receptor systems with low target tissue concentration. We investigated the ability of an optical imaging system to acquire adequate signal for in vivo measurement of receptor biochemistry. The imaging system consisted of a small animal optical imager operating in the time domain (TD) and a fluorescent-labeled diagnostic probe of known receptor-binding properties. Optical imaging of mice (n = 4) using the targeted probe, Cy5.5-DTPA galactosyl-dextran (2.2 Cy5.5, 4 DTPA, 68 galactose units per dextran, 124 kDa, 24 nmol/kg), demonstrated blood clearance and hepatic uptake. The mean and standard deviation for the time to reach 90% of the peak liver intensity were 15.4 +/- 1.6 min. Typical fluorescent intensities within a 10-pixel region-of interest from a 30-s image acquired 30 min postinjection were in excess of 2.5 million counts. The nontargeted agent (Cy5.5-DTPA-dextran) did not demonstrate (n = 4) hepatic uptake. This uptake pattern was duplicated by nuclear imaging of rabbits using (99m)Tc-labeled Cy5.5-DTPA-galactosyl-dextran and Cy5.5-DTPA dextran. This study demonstrated the feasibility of optically labeling a receptor binding diagnostic probe and imaging in the TD with sufficient sensitivity and temporal resolution for pharmacokinetic analysis. PMID- 16243644 TI - Treatment with rhenium-188-perrhenate and iodine-131 of NIS-expressing mammary cancer in a mouse model remarkably inhibited tumor growth. AB - INTRODUCTION: Novel therapeutic modalities are needed for breast cancer patients in whom standard treatments are not effective. Mammary gland sodium/iodide symporter has been identified as a molecular target in breast cancers in humans and in some transgenic mouse models. We report the results of a therapy study with (131)I(-) and (188)ReO(4)(-) of breast cancer in polyoma middle T oncoprotein (PyMT) transgenic mice endogenously expressing the Na(+)/I(-) symporter (NIS). METHODS: PyMT mice (12-13 weeks old) with one palpable tumor of 0.5-0.8 cm in diameter were used. For the therapy studies, PyMT mice were (1) treated with two intraperitoneal injections of 1.5 mCi of (188)ReO(4)(-) 1 week apart, (2) pretreated for 1 week with 5 microg of triiodothyronine (T3) followed by two intraperitoneal injections of 1.5 mCi of (131)I(-) 1 week apart or (3) left untreated. The tumor and normal organ uptakes were assessed by scintigraphic imaging. The thyroid function of treated and control animals was evaluated at the completion of the study by measuring the T3/thyroxine (T4) ratio in their blood. RESULTS: There was significant uptake of (131)I(-) and (188)ReO(4)(-) in the primary palpable tumors as well as in nonpalpable tumors, stomachs and thyroids. The tumor uptake after the second injection was 10 times lower in comparison with the first injection. Tumor growth was significantly inhibited in both the (131)I( ) and (188)ReO(4)(-) groups in comparison with the control group, and tumors in the (188)ReO(4)(-) group increased in size significantly less than in the (131)I( ) group. The T3/T4 ratios were calculated to be 27 and 25 for the control group and the (188)ReO(4)(-) group, respectively; for (131)I(-), both the T3 and T4 levels were below detection limit, demonstrating much less effect on the thyroids of treatment with (188)ReO(4)(-) than with (131)I(-). CONCLUSIONS: These results prove that NIS expression in breast tumors in animal models allows specific, efficient and safe treatment with a variety of radionuclides transported by NIS. PMID- 16243645 TI - A simple method for stem cell labeling with fluorine 18. AB - Hexadecyl-4-[(18)F]fluorobenzoate ([(18)F]HFB), a long chain fluorinated benzoic acid ester, was prepared in a one-step synthesis by aromatic nucleophilic substitution of [(18)F]fluoride ion on hexadecyl-4-(N,N,N trimethylammonio)benzoate. The radiolabeled ester was obtained in good yields (52% decay corrected) and high purity (97%). [(18)F]HFB was used to radiolabel rat mesenchymal stem cells (MSCs) by absorption into cell membranes. MicroPET imaging of [(18)F]HFB-labeled MSCs following intravenous injection into the rat showed the expected high and persistent accumulation of radioactivity in the lungs. [(18)F]HFB is thus simple to prepare and uses labeling agent for short term distribution studies of injected stem cells. PMID- 16243646 TI - New-generation radiotracers for nAChR and NET. AB - Advances in radiotracer chemistry and instrumentation have merged to make positron emission tomography (PET) a powerful tool in the biomedical sciences. Positron emission tomography has found increased application in the study of drugs affecting the brain and whole body, including the measurement of drug pharmacokinetics (using a positron-emitter-labeled drug) and drug pharmacodynamics (using a labeled tracer). Thus, radiotracers are major scientific tools enabling investigations of molecular phenomena, which are at the heart of understanding human disease and developing effective treatments; however, there is evidently a bottleneck in translating basic research to clinical practice. In the meantime, the poor ability to predict the in vivo behavior of chemical compounds based on their log P's and affinities emphasizes the need for more knowledge in this area. In this article, we focus on the development and translation of radiotracers for PET studies of the nicotinic acetylcholine receptor (nAChR) and the norepinephrine transporter (NET), two molecular systems that urgently need such an important tool to better understand their functional significance in the living human brain. PMID- 16243647 TI - Resolution, sensitivity and precision with autoradiography and small animal positron emission tomography: implications for functional brain imaging in animal research. AB - Quantitative autoradiographic methods for in vivo measurement of regional rates of cerebral blood flow, glucose metabolism, and protein synthesis contribute significantly to our understanding of phsysiological and biochemical responses of the brain to changes in the environment. A disadvantage of these autoradiographic methods is that experimental animals can be studied only once. With the advent of small animal positron emission tomography (PET) and with increases in the sensitivity and spatial resolution of scanners it is now possible to use adaptations of these methods in experimental animals with PET. These developments allow repeated studies of the same animal, including studies of the same animal under different conditions, and longitudinal studies. In this review we summarize the tradeoffs between the use of autoradiography and small animal PET for functional brain imaging studies in animal research. PMID- 16243648 TI - What is to be gained by imaging the same animal before and after treatment? AB - Experimental design includes decisions to perform measurements with intact animals or truncated in vivo measurements followed by dissection and ex vivo measurements. The design may be driven by advantages and disadvantages of the choices. Analytical results are presented here that can provide a basis for certain experimental design choices. Design considerations include longitudinal studies in the single animal that bracket a treatment, joint observations by supplementing the imaging data with plasma and/or ex vivo sampling after dissection, multiple injections of increasing molar base or administration of a molar dose that results in the occupation of a significant fraction of the target receptors. Of particular interest when constructing an experimental design is the reduction of bias. Analysis tools for this purpose include reparameterization and model application without the formation of time-based averaging. PMID- 16243649 TI - Radiolabeled peptide conjugates for targeting of the bombesin receptor superfamily subtypes. AB - Research laboratories around the world are currently focusing their efforts toward the development of radiometallated, site-directed, diagnostic/therapeutic agents based upon small peptides such as octreotide, neurotensin, alpha melanocyte stimulating hormone, vasointestinal peptide and others. Bombesin (BBN) or derivatives of bombesin are also of significant interest. Bombesin is a 14 amino-acid peptide with very high affinity for the BB2 or gastrin-releasing peptide receptor (GRPr). Over-expression of the GRPr on a variety of human cancers (i.e., breast, prostate, pancreatic, small cell lung, etc.) provides potential efficacy toward development of radiometallated BBN derivatives for targeting and, hence, diagnosis/treatment of these specific diseases. New derivatives are being developed that are also capable of targeting the BB1 and BB3 receptor subtypes that are over-expressed on cancer cells. This review highlights some of the more recent developments toward design of BBN receptor specific radiopharmaceuticals that have taken place over the past 2 years. PMID- 16243650 TI - Pretargeted radioimmunotherapy in tumored mice using an in vivo 212Pb/212Bi generator. AB - OBJECTIVE: Pretargeting is the concept that combines optimal delivery of the antibody and rapid capture and elimination of the radioactivity. In this study, we evaluated the potential of antibody pretargeting to enable the tumor-targeting (212)Pb for in vivo generation of (212)Bi for alpha particle radiotherapy. METHODS: The (212)Pb/(212)Bi chelate of DOTA-biotin, as well as their gamma emitting analogues, (203)Pb and (205)Bi, was prepared and characterized. The radiolabeled compounds were injected in animals for evaluation of tumor targeting and normal tissue uptake and retention. In the pretargeting protocol, injection of 400 microg of NR-LU-10 antibody-streptavidin conjugate was given at t = 0 h, then 100 microg of N-acetyl-galatosamine-biotin clearing agent was injected at t = 20-24 h; finally, 1 microg of (212)Pb/(212)Bi-DOTA-biotin was injected 6 h later. RESULTS: Both (203)Pb and (205)Bi-DOTA-biotin were stable for at least 4 days in the different challenging solutions including PBS, 10 mM DTPA and serum. Contrary to its gamma-emitting analogues, radiolabeled (212)Pb-DOTA-biotin was not stable. There was greater than 30% of free (212)Bi released 4 h after (212)Pb labeled DOTA-biotin. The results of pretargeting protocol of (203)Pb and (205)Bi DOTA-biotin showed that the tumor target reached 20% injected dose (ID)/g at 4 h postinjection and remained high for 5 days. The %ID/g in the whole blood and other nontarget organs was low after administration of labeled (203)Pb and (205)Bi-DOTA-biotin similar to the biodistribution of labeled DOTA-biotin alone. In the animals administered (212)Pb-DOTA-biotin, radioactivity in nontarget organs was low except the kidneys. The %ID/g in the kidney for (212)Bi was 14.5 at 2 h, higher than (212)Pb, but dropped to about 6% ID/g by 4 h. However, tumor uptake for (212)Pb and (212)Bi was >25% ID/g at 1 h postinjection and remained so through 24 h. CONCLUSIONS: Antibody pretargeting system with Mab-streptavidin, clearing agent and DOTA-biotin provides the potential of (212)Bi for solid tumor radiotherapy despite the release of (212)Bi after (212)Pb decay. Dosimetry calculations resulted in tumor dose at 93 rad/muCi and ratios of tumor to marrow and kidney at 386:1 and 12:1, respectively. PMID- 16243651 TI - Gene manipulation to enhance MIBG-targeted radionuclide therapy. AB - The goal of targeted radionuclide therapy is the deposition in malignant cells of sterilizing doses of radiation without damaging normal tissue. The radiopharmaceutical [(131)I]meta-iodobenzylguanidine ([(131)I]MIBG) is an effective single agent for the treatment of neuroblastoma. However, uptake of the drug in malignant sites is insufficient to cure disease. A growing body of experimental evidence indicates exciting possibilities for the integration of gene transfer with [(131)I]MIBG-targeted radiotherapy. PMID- 16243652 TI - Positron emission tomography studies in eating disorders: multireceptor brain imaging, correlates with behavior and implications for pharmacotherapy. AB - Modern imaging techniques that visualize disease-specific organ neurotransmitter or protein receptor sites are increasingly able to define pathological processes on a molecular level. One of those imaging modalities, positron emission tomography (PET), for the assessment of brain neuroreceptor binding has revolutionized the in vivo assessment of biologic markers that may be related to human behavior. Such studies may help identify chemical targets that may be directly related to psychiatric pathology and, thus, opportunities for pharmacological intervention. In this review, we describe results from PET studies in eating disorders (EDs). Eating disorders are frequently debilitating illnesses that are quite homogeneous in their presentation. Those studies that identified particular serotonin and dopamine receptor alterations can distinguish recovered ED subjects from controls as well as ED subgroups. Furthermore, correlations of receptor binding with behavioral constructs, such as harm avoidance or novelty seeking, could be found. These recognized receptors may now help us to move away from rather nonspecific treatment approaches in psychiatric research and clinic to the possibility of more syndrome- and symptom-specific treatment approaches. PMID- 16243653 TI - Reporter gene imaging: potential impact on therapy. AB - Positron emission tomography (PET)-based molecular-genetic imaging in living organisms has enjoyed exceptional growth over the past 5 years; this is particularly striking since it has been identified as a new discipline only within the past decade. Positron emission tomography is one of three imaging technologies (nuclear, magnetic resonance and optical) that has begun to incorporate methods that are established in molecular and cell biology research. The convergence of these disciplines and the wider application of multi-modality imaging are at the heart of this success story. Most current molecular-genetic imaging strategies are "indirect," coupling a "reporter gene" with a complimentary "reporter probe." Reporter gene constructs can be driven by constitutive promoter elements and used to monitor gene therapy vectors and the efficacy of trans gene targeting and transduction, as well as to monitor adoptive cell-based therapies. Inducible promoters can be used as "sensors" to regulate the magnitude of reporter gene expression and can be used to provide information about endogenous cell processes. Reporter systems can also be constructed to monitor mRNA stabilization and specific protein-protein interactions. Promoters can be cell specific and restrict transgene expression to certain tissue and organs. The translation of reporter gene imaging to specific clinical applications is discussed. Several examples that have potential for patient imaging studies in the near future include monitoring adenoviral-based gene therapy, oncolytic herpes virus therapy, adoptive cell-based therapies and Salmonella-based tumor-targeted cancer therapy and imaging. The primary translational applications of noninvasive in vivo reporter gene imaging are likely to be (a) quantitative monitoring of the gene therapy vector and the efficacy of transduction in clinical protocols, by imaging the location, extent and duration of transgene expression; (b) monitoring cell trafficking, targeting, replication and activation in adoptive therapies, involving ex vivo transduction of harvested immune-competent cells and stem/progenitor cells; (c) assessments of endogenous molecular events using different reporter gene imaging technologies following the development of safe, efficient and target-specific vectors for "diagnostic transductions." PMID- 16243654 TI - Breast imaging: a surgeon's prospective. AB - Mammography, ultrasound, magnetic resonance imaging, positron emission tomography, gamma camera and intraoperative gamma detection, and computed tomography are employed in the diagnosis and treatment of breast cancer. This paper summarizes the role of each modality from the perspective of the physician responsible for management of the patient's care. An understanding of an imaging modality's current role can provide insights into the design of new applications and diagnostic agents. Moreover, knowledge of the mechanism by which each modality provides clinical information can guide the design of new imaging methods that complement and add certainty to the patient's management. The reader should note the lack of molecular information provided by the current imaging methods. The perspective concludes with a request for an imaging technique that can measure the biologic aggressiveness of a woman's cancer. The surgeon notes that basing the formation of an image on a molecular process would be compatible with current medical practice, which utilizes molecular concepts to base medical decisions. In addition, molecular imaging will enable rapid translation between basic science and medical practice. PMID- 16243657 TI - Transplant case manager is perplexed and confused. PMID- 16243659 TI - Integrating CAM into health care professional education. PMID- 16243660 TI - Case management: heart and soul. AB - Dear Colleagues: As a seasoned case manager, I have had the opportunity to serve our specialty practice in the development of the definition of case management, the creation of our standards of practice, the evolution of our professional code of conduct, and, of course, the process of certification. Yet it is the stories of our shared time with our clients that truly define what we do and who we are and clarify our contribution to health care. It is by the telling of the story that we understand our roles, our behavior, and the value of our activities. PMID- 16243661 TI - Managing resistance to change: the social worker's role in case management. AB - From time to time in my responsibilities as a medical social worker, I become involved with a "complicated" case. Translation: a case was referred to the social worker to intervene and "fix" the problem because no one else knew what to do with the client or the issues surrounding the client. Necessity is the mother of invention, so when all else failed, that was my cue to be creative. PMID- 16243662 TI - Performance measurement for case management: principles and objectives for developing standard measures. AB - Developing standardized performance measurements for case management (CM) has become the holy grail of the field. The Council for Case Management Accountability (CCMA), a leadership committee of the Case Management Society of America (CMSA), has been grappling with the concept since early 2003. This article lays out a theoretical framework for performance measurement and then outlines the progress on a specific initiative begun by CCMA to correlate CM practices with improved outcome measures. PMID- 16243663 TI - Consumer-directed health care and case management: Part 2. PMID- 16243664 TI - Converging pathways: a journey toward quality case management. AB - Development of case management as a profession surged in the latter part of the last century, due in large part to the birth in 1990 of two quality-focused organizations. In that year, URAC was established to improve the quality and the accountability of organizations conducting utilization review, but, in later years, it expanded to accreditation of organizations conducting many medical management components, such as case management and disease management. Today the organization has 17 accreditation programs. PMID- 16243665 TI - Cuffed endotracheal tubes. PMID- 16243667 TI - Diabetic ketoacidosis in the pediatric population. PMID- 16243673 TI - The new two-patient care environment. PMID- 16243674 TI - Austin-Travis County STAR Flight. PMID- 16243676 TI - Appropriateness of medical transport and access to care in acute stroke syndromes. PMID- 16243675 TI - Off-duty preparation for overnight work in rotor wing air medical programs. AB - INTRODUCTION: Air medical teams provide around-the-clock critical care, risking performance-altering fatigue from circadian disruption and sleep deprivation. Although safety is an essential issue in the air medical industry, there is little understanding of off-duty preparation for overnight shifts. METHODS: An anonymous survey was distributed to pilots and medical team members at participating programs with variable program, staffing, and shift models. Eighty responses from crewmembers working 12-hour night shifts (12N) were analyzed with appropriate t-tests and nonparametric tests. RESULTS: 12N crewmembers sleep significantly less in off-duty periods than before night shifts: 7.3 +/- 1.2 hours versus 4.8 +/- 1.9 hours (P < 0.01). Preshift sleep does not differ between crewmembers permitted on-duty rest and those for whom it is prohibited. 34.1% of 12N crewmembers permitted on-duty rest say they report to work planning to sleep. The minimum preshift sleep reported by 12N crewmembers before any shift in the past month averaged 2.4 +/- 2.3 hours, with 36.3% having worked overnight in the past month with no sleep before their shift On-duty rest permission was not a significant factor. Fifty-five percent of 12N crewmembers report outside employment (OE) in addition to their flight position. 12N crewmembers with OE averaged significantly less preshift sleep than those without OE: 4.4 +/- 2.1 hours versus 5.3 +/- 1.6 hours (P < 0.05). 54.5% of 12N crewmembers with OE described reporting to a flight shift within 8 hours of leaving their other job at least once within the past month. OE was more common when the flight program permitted on-duty rest (P < 0.01). CONCLUSION: Air medical team members report for 12N shifts with a significant sleep debt that does not differ between crewmembers permitted on-duty rest and those with on-duty rest prohibitions. More than half of flight team members surveyed have OE and many report for flight duty within 8 hours of leaving their other job. 12N shift crewmembers are at a particularly high risk for the consequences of fatigue. This is an important consideration as the industry develops on-duty rest guidelines to optimize safe operations. PMID- 16243677 TI - The multi-dimensional neighbourhood and health: a cross-sectional analysis of the Scottish Household Survey, 2001. AB - Neighbourhoods may influence the health of individual residents in different ways: via the social and physical environment, as well as through facilities and services. Not all factors may be equally important for all population subgroups. A cross-sectional analysis of the Scottish Household Survey 2001 examined a range of neighbourhood factors for links with three health outcomes and two health related behaviours. The results support the hypothesis that the neighbourhood has a multi-dimensional impact on health. There was also some evidence that the relationship between neighbourhood factors and health varied according to the population subgroup, although not in a consistent manner. PMID- 16243678 TI - The ecological relationship between deprivation, social isolation and rates of hospital admission for acute psychiatric care: a comparison of London and New York City. AB - We report on comparative analyses of small area variation in rates of acute hospital admissions for psychiatric conditions in Greater London around the year 1998 and in New York City (NYC) in 2000. Based on a theoretical model of the factors likely to influence psychiatric admission rates, and using data from the most recent population censuses and other sources, we examine the association with area indicators designed to measure access to hospital beds, socio-economic deprivation, social fragmentation and ethnic/racial composition. We report results on admissions for men and women aged 15-64 for all psychiatric conditions (excluding self-harm), drug-related substance abuse/addiction, schizophrenia and affective disorders. The units of analysis in NYC were 165 five-digit Zip Code Areas and, in London, 760 electoral wards as defined in 1998. The analysis controls for age and sex composition and, as a proxy for access to care, spatial proximity to hospitals with psychiatric beds. Poisson regression modeling incorporating random effects was used to control for both overdispersion in the counts of admissions and for the effects of spatial autocorrelation. The results for NYC and London showed that local admission rates for all types of condition were positively and significantly associated with deprivation and the association is independent of demographic composition or 'access' to beds. In NYC, social fragmentation showed a significant association with admissions due to affective disorders and schizophrenia, and for drug dependency among females. Racial minority concentration was significantly and positively associated with admissions for schizophrenia. In London, social fragmentation was associated positively with admissions for men and women due to schizophrenia and affective disorders. The variable measuring racial/ethnic minority concentration for London wards showed a negative association with admission rates for drug dependency and for affective disorders. We discuss the interpretation of these results and the issues they raise in terms of the potential and limitations of international comparison. PMID- 16243679 TI - Ecological analysis of the distribution and socio-spatial context of homicides in Porto Alegre, Brazil. AB - Over the last decade, the number of homicides in Porto Alegre has increased to the point where external causes are now the main group of causes of death in the 5-34-year age group. Preventing these deaths depends fundamentally on identifying factors related to excess violence in population groups. The overall aim of this study is to analyse the spatial distribution of homicide victims by place of residence in Porto Alegre, the capital of the southernmost Brazilian State of Rio Grande do Sul, in 1996, in order to identify and understand the socio-spatial context. Demographic and socioeconomic indicators based on the 1991 census and 1996 population count were used to build a multivariate classification characterizing the 1851 census tracts. Homicides occurring in 1996 were located using the municipality's Geographic Information System. Four socioeconomic groups were identified, mainly differentiated by housing indicators. Small areas on the urban periphery in which slums (favelas) are concentrated presented higher homicide rates. Homicide rates were lower in the two groups with higher income and educational level. The second step was to classify the census tracts according to the homicide indicator. In this case, areas were differentiated by the number of household inhabitants per room, income, schooling, and median age. We conclude that the multivariate socioeconomic classification presents a limited capacity to identify populations exposed to homicides, suggesting that socioeconomic conditions themselves do not determine violent behaviour. On the other hand, the spatial methods allowed us to identify small areas where deaths are concentrated and whose populations should receive special attention in planning measures to prevent violent deaths. PMID- 16243680 TI - Intra-urban differentials in child dental trauma in relation to healthy cities policies in Curitiba, Brazil. AB - This paper is concerned with developing methods for assessing the potential impact of healthy public policies on health outcomes, particularly on dental trauma. The hypothesis is that in deprived areas of Curitiba, Brazil, there would be a lower prevalence of dental trauma in 12-year-old schoolchildren related to different degrees of implementation of healthy public policies. Fourteen socio environmental variables gathered from 29 deprived areas were factor-analysed and three principal components (PCA) extracted: physical environment, public social policies, and social cohesion. Individual clinical data on dental trauma were collected for 2,126 children from public schools in the areas. Rank correlation indicated that the relationship between components' scores and dental trauma was statistically significant. The PCA scores explained 42% of the variance for dental trauma. PMID- 16243681 TI - Inscribing healthification: governance, risk, surveillance and the subjects and spaces of fitness and health. AB - This paper considers the interface between the hygienic geographies of a fitness and exercise space and the discourses of risk and subjectivity in this era of the new public health. Using an analysis of space, power and the subject, the paper assesses the ways in which subjects govern themselves and others in public health spaces through an intensification of surveillance, in order to ensure health and safety compliance. In this paper, first, I introduce the locker room as a place of health and hygiene. Second, I set out the wider context of the inscription of health on spaces and subjects. Third, I examine the relationship between discourses of risk and subjectivity and how risk discourses are fundamental to the fabrication of subjects and social life. The paper then examines, more specifically, techniques of self- and other-governance that help to inscribe and prescribe the new public health in locker rooms. Drawing on the findings of a spatial and ethnographic case study of an urban, Canadian university locker room, I conclude that the macro- and micro-politics and economics of policing public health require both the regulation of bodies and the participation of active, health-conscious citizens in order to keep (western) neo-liberal imperatives of health in place. PMID- 16243682 TI - Unstable housing, associated risk behaviour, and increased risk for HIV infection among injection drug users. AB - We sought to examine the relationship between housing status and risk of HIV infection among injection drug users in Vancouver, Canada. Using Kaplan-Meier survival analysis, we found an elevated HIV incidence rate among those who reported residing in unstable housing (log-rank p=0.006). In Cox's regression survival analysis, unstable housing remained marginally associated with elevated risks of HIV infection (relative hazard=1.40 (95% confidence interval: 0.09 2.00); p=0.084) after adjustment for potential confounders including syringe sharing. Adjusted generalized estimating equations analysis that examined factors associated with unstable housing demonstrated that residing in unstable housing was independently associated with several HIV risk behaviours including borrowing used needles (adjusted odds ratio (OR)=1.14) and sex-trade involvement (adjusted OR=1.19). Our findings suggest that unstable housing environments are associated with elevated risk of HIV- infection due to risk behaviours that take place in these environments. Implications for policy including more comprehensive housing interventions (e.g. 'floating support') are discussed. PMID- 16243684 TI - Levelling the playing field? Exploring the implications of managed competition for voluntary sector providers of long-term care in small town Ontario. AB - This paper considers how long-term care restructuring, under the rubric of managed competition, has increased pressure on voluntary sector providers of long term care to elderly populations in small, rural places. Drawing on in-depth interviews with key-informants from non-profit organizations, we present a case study set in rural Ontario to develop a situated understanding of the impact of restructuring on voluntary sector providers of long-term care. We contend that managed competition (underway in Ontario since 1995) has constrained providers, eroded service choices, and reduced access to long-term care in rural areas leaving rural populations more disadvantaged than ever before when it comes to public service provisioning. PMID- 16243683 TI - Pokies and poverty: problem gambling risk factor geography in New Zealand. AB - Up to 2% of adults in New Zealand can be considered problem gamblers, where the activity has an impact on the well-being of those who gamble, and often their close associates. The most common activity involves non-casino gaming machines (NCGMs). This paper explores the geography of gambling 'opportunity' at small area scales, and finds excess provision in those areas classified as highly deprived. Geographically weighted regression has been used to investigate the possibility that the degree of inequity in NCGM provision varies across New Zealand. As machines are licensed, this provides an opportunity for policy implementation towards risk reduction. PMID- 16243685 TI - Geographies of health in nursing. PMID- 16243686 TI - Rodent malaria parasites Plasmodium chabaudi and P. vinckei do not increase their rates of gametocytogenesis in response to mosquito probing. AB - Several vector-borne infectious agents facultatively alter their life history strategies in response to local vector densities. Some evidence suggests that malaria parasites invest more heavily in transmission stage production (gametocytogenesis) when vectors are present. Such a strategy could rapidly increase malaria transmission rates, particularly when adult mosquitoes begin to appear after dry seasons. However, in contrast to a recent experiment with a rodent malaria (Plasmodium chabaudi), we found no change in gametocytogenesis in either P. chabaudi or in another rodent malaria, P. vinckei, when their mouse hosts were exposed to mosquitoes. Positive results in the earlier study may have been because mosquito-feeding caused anaemia in hosts, a known promoter of gametocytogenesis. The substantial evidence that malaria and a variety of other parasites facultatively alter transmission strategies in response to a variety of environmental influences makes our results surprising. PMID- 16243687 TI - Do unprofitable prey evolve traits that profitable prey find difficult to exploit? AB - Prey that are unprofitable to attack (for example, those containing noxious chemicals) are often conspicuously patterned and move in a slower and more predictable manner than species lacking these defences. Contemporary theories suggest these traits have evolved as warning signals because they can facilitate both associative and discriminative avoidance learning in predators. However, it is unclear why these particular traits and not others have tended to evolve in unprofitable prey. Here we show using a signal detection model that unprofitable prey will evolve conspicuous colours and patterns partly because these characteristics cannot readily evolve in profitable prey without close mimicry. The stability of this signal is maintained through the costs of dishonesty in profitable prey. Indeed, unprofitable prey will sometimes evolve a conspicuous form to reduce mimetic parasitism, even in the unlikely event that this form can be more closely mimicked. This is one of the first mathematical models of the evolution of warning signals to allow for the possibility of mimicry, yet our analyses suggest it may offer a general explanation as to why warning signals take the form that they do. Warning signals and mimicry may therefore be more closely related than is currently supposed. PMID- 16243689 TI - Recurrent evolution of host-specialized races in a globally distributed parasite. AB - The outcome of coevolutionary interactions is predicted to vary across landscapes depending on local conditions and levels of gene flow, with some populations evolving more extreme specializations than others. Using a globally distributed parasite of colonial seabirds, the tick Ixodes uriae, we examined how host availability and geographic isolation influences this process. In particular, we sampled ticks from 30 populations of six different seabird host species, three in the Southern Hemisphere and three in the Northern Hemisphere. We show that parasite races have evolved independently on hosts of both hemispheres. Moreover, the degree of differentiation between tick races varied spatially within each region and suggests that the divergence of tick races is an ongoing process that has occurred multiple times across isolated areas. As I. uriae is vector to the bacterium responsible for Lyme disease Borrelia burgdorferi sensu lato, these results may have important consequence for the epidemiology of this disease. With the increased occurrence of novel interspecific interactions due to global change, these results also stress the importance of the combined effects of gene flow and selection for parasite diversification. PMID- 16243688 TI - Socially induced and rapid increases in aggression are inversely related to brain aromatase activity in a sex-changing fish, Lythrypnus dalli. AB - Social interactions can generate rapid and dramatic changes in behaviour and neuroendocrine activity. We investigated the effects of a changing social environment on aggressive behaviour and brain aromatase activity (bAA) in a sex changing fish, Lythrypnus dalli. Aromatase is responsible for the conversion of androgen into oestradiol. Male removal from a socially stable group resulted in rapid and dramatic (> or =200%) increases in aggression in the dominant female, which will become male usually 7-10 days later. These dominant females and recently sex-changed individuals had lower bAA but similar gonadal aromatase activity (gAA) compared to control females, while established males had lower bAA than all groups and lower gAA than all groups except dominant females. Within hours of male removal, dominant females' aggressive behaviour was inversely related to bAA but not gAA. These results are novel because they are the first to: (i) demonstrate socially induced decreases in bAA levels corresponding with increased aggression, (ii) identify this process as a possible neurochemical mechanism regulating the induction of behavioural, and subsequently gonadal, sex change and (iii) show differential regulation of bAA versus gAA resulting from social manipulations. Combined with other studies, this suggests that aromatase activity may modulate fast changes in vertebrate social behaviour. PMID- 16243690 TI - Does Bertrand's rule apply to macronutrients? AB - It has been known for over a century that the dose-response curve for many micronutrients is non-monotonic, having an initial stage of increasing benefits with increased intake, followed by increasing costs as excesses become toxic. This phenomenon, termed Bertrand's rule, is widely assumed not to apply to caloric macronutrients. To date this assumption has been safe, owing to the considerable methodological challenges involved in coaxing animals to over-ingest macronutrients in a way that enables the effects of specific food components to be isolated. Here we report an experiment which overcomes these difficulties, to test whether the second phase (incurring costs with excessive intake) applies to carbohydrate intake by the generalist-feeding caterpillar Spodoptera littoralis. The results showed that excess carbohydrate intake caused increased mortality, thus extending Bertrand's rule to macronutrients. PMID- 16243691 TI - Winter resource wealth drives delayed dispersal and family-group living in western bluebirds. AB - Delayed dispersal, where offspring remain with parents beyond the usual period of dependence, is the typical route leading to formation of kin-based cooperative societies. The prevailing explanations for why offspring stay home are variation in resource wealth, in which offspring of wealthy parents benefit disproportionately by staying home, and nepotism, where the tendency for parents to be less aggressive and share food with offspring makes home a superior place to wait to breed. These hypotheses are not strict alternatives, as only wealthy parents have sufficient resources to share. In western bluebirds, Sialia mexicana, sons usually delay dispersal until after winter, gaining feeding advantages through maternal nepotism in a familial winter group. Experimentally reducing resource wealth (mistletoe) by half on winter territories caused sons to disperse in summer, even though their parents remained on the territory during the winter. Only 8% of sons remained with their parents on mistletoe-removal territories compared to 50% of sons on control territories (t(9,10)=3.33, p<0.005). This study is the first to demonstrate that experimentally reducing wealth of a natural food resource reduces delayed dispersal, facilitating nepotism and family-group living. The results clarify the roles of year-round residency, resource limitation and relative wealth outside the breeding season in facilitating the formation of kin-based cooperative societies. PMID- 16243692 TI - Modelling information exchange in worker-queen conflict over sex allocation. AB - We investigate the conflict between queen and worker over sex allocation, specifically the allocation of the queen's eggs between workers and reproductives and the allocation of the reproductive eggs between male and female. In contrast to previous models, we allow workers to observe and use information about the strategy of the queen. We consider three conflict models: simultaneous (no information exchange), sequential (a one-way information exchange) and negotiated (an iterated two-way information exchange). We find that the first model produces sex ratios intermediate between the classic queen (1:1) and worker (1:3) optima. The second model, in which the worker has information about the queen's decisions, produces a different result and one that is somewhat counter-intuitive in that the sex ratios are less female-biased than for the other two models, and in fact are often male-biased. The third model predicts sex ratios intermediate between the first two models. We discuss how these findings may shed new light on observed sex allocation patterns in social insects and we suggest some experimental tests. PMID- 16243693 TI - Cattle domestication in the Near East was followed by hybridization with aurochs bulls in Europe. AB - Domesticated cattle were one of the cornerstones of European Neolithisation and are thought to have been introduced to Europe from areas of aurochs domestication in the Near East. This is consistent with mitochondrial DNA (mtDNA) data, where a clear separation exists between modern European cattle and ancient specimens of British aurochsen. However, we show that Y chromosome haplotypes of north European cattle breeds are more similar to haplotypes from ancient specimens of European aurochsen, than to contemporary cattle breeds from southern Europe and the Near East. There is a sharp north-south gradient across Europe among modern cattle breeds in the frequencies of two distinct Y chromosome haplotypes; the northern haplotype is found in 20 out of 21 European aurochsen or early domestic cattle dated 9500-1000 BC. This indicates that local hybridization with male aurochsen has left a paternal imprint on the genetic composition of modern central and north European breeds. Surreptitious mating between aurochs bulls and domestic cows may have been hard to avoid, or may have occurred intentionally to improve the breeding stock. Rather than originating from a few geographical areas only, as indicated by mtDNA, our data suggest that the origin of domestic cattle may be far more complex than previously thought. PMID- 16243694 TI - Odour concentration affects odour identity in honeybees. AB - The fact that most types of sensory stimuli occur naturally over a large range of intensities is a challenge to early sensory processing. Sensory mechanisms appear to be optimized to extract perceptually significant stimulus fluctuations that can be analysed in a manner largely independent of the absolute stimulus intensity. This general principle may not, however, extend to olfaction; many studies have suggested that olfactory stimuli are not perceptually invariant with respect to odour intensity. For many animals, absolute odour intensity may be a feature in itself, such that it forms a part of odour identity and thus plays an important role in discrimination alongside other odour properties such as the molecular identity of the odorant. The experiments with honeybees reported here show a departure from odour-concentration invariance and are consistent with a lower-concentration regime in which odour concentration contributes to overall odour identity and a higher-concentration regime in which it may not. We argue that this could be a natural consequence of odour coding and suggest how an 'intensity feature' might be useful to the honeybee in natural odour detection and discrimination. PMID- 16243695 TI - A ratio model of perceived speed in the human visual system. AB - The perceived speed of moving images changes over time. Prolonged viewing of a pattern (adaptation) leads to an exponential decrease in its perceived speed. Similarly, responses of neurones tuned to motion reduce exponentially over time. It is tempting to link these phenomena. However, under certain conditions, perceived speed increases after adaptation and the time course of these perceptual effects varies widely. We propose a model that comprises two temporally tuned mechanisms whose sensitivities reduce exponentially over time. Perceived speed is taken as the ratio of these filters' outputs. The model captures increases and decreases in perceived speed following adaptation and describes our data well with just four free parameters. Whilst the model captures perceptual time courses that vary widely, parameter estimates for the time constants of the underlying filters are in good agreement with estimates of the time course of adaptation of direction selective neurones in the mammalian visual system. PMID- 16243696 TI - Biology of extinction risk in marine fishes. AB - We review interactions between extrinsic threats to marine fishes and intrinsic aspects of their biology that determine how populations and species respond to those threats. Information is available on the status of less than 5% of the world's approximately 15500 marine fish species, most of which are of commercial importance. By 2001, based on data from 98 North Atlantic and northeast Pacific populations, marine fishes had declined by a median 65% in breeding biomass from known historic levels; 28 populations had declined by more than 80%. Most of these declines would be sufficient to warrant a status of threatened with extinction under international threat criteria. However, this interpretation is highly controversial, in part because of a perception that marine fishes have a suite of life history characteristics, including high fecundity and large geographical ranges, which might confer greater resilience than that shown by terrestrial vertebrates. We review 15 comparative analyses that have tested for these and other life history correlates of vulnerability in marine fishes. The empirical evidence suggests that large body size and late maturity are the best predictors of vulnerability to fishing, regardless of whether differences among taxa in fishing mortality are controlled; there is no evidence that high fecundity confers increased resilience. The evidence reviewed here is of direct relevance to the diverse criteria used at global and national levels by various bodies to assess threat status of fishes. Simple life history traits can be incorporated directly into quantitative assessment criteria, or used to modify the conclusions of quantitative assessments, or used as preliminary screening criteria for assessment of the approximately 95% of marine fish species whose status has yet to be evaluated either by conservationists or fisheries scientists. PMID- 16243697 TI - Metamorphosis in a Silurian barnacle. AB - Exceptionally preserved fossils from the Wenlock Series (Silurian) of Herefordshire, UK, provide unique evidence of metamorphosis from free-swimming cyprid larva to attached juvenile in a Palaeozoic barnacle. The larva had large brush-like anterior limbs. The juvenile shows the head transformed into a stalk and the development of the primordial condition of five mineralized plates within the carapace. The discovery of a cyprid larva indicates that crown group cirripedes had evolved by the Silurian. PMID- 16243698 TI - An invasive lineage of sculpins, Cottus sp. (Pisces, Teleostei) in the Rhine with new habitat adaptations has originated from hybridization between old phylogeographic groups. AB - Fish abundance surveys in the Rhine system have shown in the past two decades that there is a rapid upriver invasion of a freshwater sculpin of the genus Cottus. These fish are found in habitats that are atypical for the known species Cottus gobio, which is confined to small cold streams within the Rhine drainage. Phylogeographic analysis based on mitochondrial haplotypes and diagnostic single nucleotide polymorphisms indicates that the invasive sculpins are hybrids between two old lineages from the River Scheldt drainage and the River Rhine drainage, although it is morphologically more similar to the Scheldt sculpins. Most importantly, however, the invasive population possesses a unique ecological potential that does not occur in either of the source populations from the Rhine or the Scheldt, which allows the colonization of new habitats that have previously been free of sculpins. Microsatellite analysis shows that the new lineage is genetically intermediate between the old lineages and that it forms a distinct genetic group across its whole expansion range. We conclude that hybridization between long separated groups has lead to the fast emergence of a new, adaptationally distinct sculpin lineage. PMID- 16243699 TI - Genetic analysis reveals demographic fragmentation of grizzly bears yielding vulnerably small populations. AB - Ecosystem conservation requires the presence of native carnivores, yet in North America, the distributions of many larger carnivores have contracted. Large carnivores live at low densities and require large areas to thrive at the population level. Therefore, if human-dominated landscapes fragment remaining carnivore populations, small and demographically vulnerable populations may result. Grizzly bear range contraction in the conterminous USA has left four fragmented populations, three of which remain along the Canada-USA border. A tenet of grizzly bear conservation is that the viability of these populations requires demographic linkage (i.e. inter-population movement of both sexes) to Canadian bears. Using individual-based genetic analysis, our results suggest this demographic connection has been severed across their entire range in southern Canada by a highway and associated settlements, limiting female and reducing male movement. Two resulting populations are vulnerably small (< or =100 animals) and one of these is completely isolated. Our results suggest that these trans-border bear populations may be more threatened than previously thought and that conservation efforts must expand to include international connectivity management. They also demonstrate the ability of genetic analysis to detect gender-specific demographic population fragmentation in recently disturbed systems, a traditionally intractable yet increasingly important ecological measurement worldwide. PMID- 16243700 TI - Molecular evolution of the sheep prion protein gene. AB - Transmissible spongiform encephalopathies (TSEs) are infectious, fatal neurodegenerative diseases characterized by aggregates of modified forms of the prion protein (PrP) in the central nervous system. Well known examples include variant Creutzfeldt-Jakob Disease (vCJD) in humans, BSE in cattle, chronic wasting disease in deer and scrapie in sheep and goats. In humans, sheep and deer, disease susceptibility is determined by host genotype at the prion protein gene (PRNP). Here I examine the molecular evolution of PRNP in ruminants and show that variation in sheep appears to have been maintained by balancing selection, a profoundly different process from that seen in other ruminants. Scrapie eradication programs such as those recently implemented in the UK, USA and elsewhere are based on the assumption that PRNP is under positive selection in response to scrapie. If, as these data suggest, that assumption is wrong, eradication programs will disrupt this balancing selection, and may have a negative impact on the fitness or scrapie resistance of national flocks. PMID- 16243701 TI - Importance of climatological downscaling and plant phenology for red deer in heterogeneous landscapes. AB - Understanding how climate influences ecosystems represents a challenge in ecology and natural resource management. Although we know that climate affects plant phenology and herbivore performances at any single site, no study has directly coupled the topography-climate interaction (i.e. the climatological downscaling process) with large-scale vegetation dynamics and animal performances. Here we show how climatic variability (measured by the North Atlantic oscillation 'NAO') interacts with local topography in determining the vegetative greenness (as measured by the normalized difference vegetation index 'NDVI') and the body masses and seasonal movements of red deer (Cervus elaphus) in Norway. Warm springs induced an earlier onset of vegetation, resulting in earlier migration and higher body masses. Increasing values of the winter-NAO corresponded to less snow at low altitude (warmer, more precipitation results in more rain), but more snow at high altitude (colder, more precipitation corresponds to more snow) relative to winters with low winter-NAO. An increasing NAO thus results in a spatially more variable phenology, offering migrating deer an extended period with access to high-quality forage leading to increased body mass. Our results emphasize the importance of incorporating spring as well as the interaction between winter climate and topography when aiming at understanding how plant and animal respond to climate change. PMID- 16243702 TI - Thermodynamics in solid mechanics: a commentary. AB - This commentary on thermodynamics in solid mechanics aims to provide an overview of the main concepts of thermodynamic processes as they apply to, and may be exploited for, studies in nonlinear solid mechanics. We give a descriptive commentary on the (physical) interpretation of these concepts, and relate these where appropriate to behaviour of solids under thermo-mechanical conditions. The motivation is firstly that students of solid mechanics have often had less exposure to thermodynamics than those in other branches of science and engineering, yet there is great value in analytical formulations of material behaviour derived from the principles of thermodynamics. It also sets the contributions in this Theme Issue in context. Along with the deliberately descriptive treatment of thermodynamics, we do outline the main mathematical statements that define the subject, knowing that full details are provided by the authors in their corresponding contributions to this issue. The commentary ends on a lighter note. In order to aid understanding and to stimulate discussion of thermodynamics in solid mechanics, we have invented a number of very basic and completely fictitious materials. These have strange and extreme behaviours that describe certain thermodynamics concepts, such as entropy, in isolation from the complexities of real material behaviour. PMID- 16243703 TI - An anisotropic thermomechanical damage model for concrete at transient elevated temperatures. AB - The behaviour of concrete at elevated temperatures is important for an assessment of integrity (strength and durability) of structures exposed to a high temperature environment, in applications such as fire exposure, smelting plants and nuclear installations. In modelling terms, a coupled thermomechanical analysis represents a generalization of the computational mechanics of fracture and damage. Here, we develop a fully coupled anisotropic thermomechanical damage model for concrete under high stress and transient temperature, with emphasis on the adherence of the model to the laws of thermodynamics. Specific analytical results are given, deduced from thermodynamics, of a novel interpretation on specific heat, evolution of entropy and the identification of the complete anisotropic, thermomechanical damage surface. The model is also shown to be stable in a computational sense, and to satisfy the laws of thermodynamics. PMID- 16243704 TI - Thermodynamically consistent relations involving plasticity, internal energy and thermal effects. AB - Experimental data associated with plastic deformations indicate that the temperature is less than that predicted from dissipation based on plastic work. To obtain reasonable correlation between theoretical and experimental results, the plastic work is often multiplied by a constant beta. This paper provides an alternative thermodynamic framework in which it is proposed that there is an additional internal energy associated with dislocation pile-up or increase in dislocation density. The form of this internal energy follows from experimental data that relates flow stress to dislocation density and to equivalent plastic strain. The result is that beta is not a constant but a derived function. Representative results for beta and temperature as functions of effective plastic strain are provided for both an uncoupled and a coupled thermoplastic theory. In addition to providing features that are believed to be representative of many metals, the formulation can be used as a basis for more advanced theories such as those needed for large deformations and general forms of internal energy. PMID- 16243705 TI - Micromechanics investigation of expansive reactions in chemoelastic concrete. AB - Expansive reactions damage porous materials through the formation of reaction products of a volume in excess of the available space left by the reactants and the natural porosity of the material. This leads to pressurizing the pore space accessible to the reaction products, which differs when the chemical reaction is through-solution or topochemical or both in nature. This paper investigates expansive reactions from a micromechanical point of view, which allows bridging the scale from the local chemo-mechanical mechanisms to the macroscopically observable stress-free expansion. In particular, the study of the effect of morphology of the pore space, in which the chemical expansion occurs locally, on the macroscopically observable expansion is the main focus of this paper. The first part revisits the through-solution and the topochemical reaction mechanism within the framework of micro-macro-homogenization theories, and the effect of the microscopic geometry of pores and microcracks in the solid matrix on the macroscopic chemical expansion is examined. The second part deals with the transition from a topochemical to a through-solution-like mechanism that occurs in a solid matrix with inclusions (cracks, pores) of different morphology. PMID- 16243706 TI - Thermodynamic conditions for stability in materials with rate-independent dissipation. AB - A distinctive feature of the examined class of solids is that a part of the entropy production is due to rate-independent dissipation, as in models of plasticity, damage or martensitic transformations. The standard condition for thermodynamic stability is shown to be too restrictive for such solids and, therefore, an extended condition for stability of equilibrium is developed. The classical thermodynamic theory of irreversible processes is used along with the internal variable approach, with the emphasis on the macroscopic effects of micro scale instabilities in the presence of two different scales of time. Specific conditions for material stability against internal structural rearrangements under deformation-sensitive loading are derived within the incremental constitutive framework of multi-mode inelasticity. Application to spontaneous formation of deformation bands in a continuum is presented. Conditions for stability or instability of a quasi-static process induced by varying loading are given under additional constitutive postulates of normality and symmetry. As illustration of the theory, the stability of equilibrium or a deformation path under uniaxial tension is analysed for a class of inelastic constitutive laws for a metal crystal deformed plastically by multi-slip or undergoing stress-induced martensitic transformation. PMID- 16243707 TI - Enhanced continua and discrete lattices for modelling granular assemblies. AB - This article discusses the derivation of continuum models that can be used for modelling the inhomogeneous mechanical behaviour of granular assemblies. These so called kinematically enhanced models are of the strain-gradient type and of the strain-gradient micro-polar type, and are derived by means of homogenizing the micro-structural interactions between discrete particles. By analysis of the body wave dispersion curves, the enhanced continuum models are compared to corresponding discrete lattice models. Accordingly, it can be examined up to which deformation level the continuum models are able to accurately describe the discrete particle behaviour. Further, the boundary conditions for the enhanced continuum models are formulated, and their stability is considered. It is demonstrated how to use the body wave dispersion relations for the assessment of stability. PMID- 16243708 TI - High-temperature creep rupture of low alloy ferritic steel butt-welded pipes subjected to combined internal pressure and end loadings. AB - Constitutive equations are reviewed and presented for low alloy ferritic steels which undergo creep deformation and damage at high temperatures; and, a thermodynamic framework is provided for the deformation rate potentials used in the equations. Finite element continuum damage mechanics studies have been carried out using these constitutive equations on butt-welded low alloy ferritic steel pipes subjected to combined internal pressure and axial loads at 590 and 620 degrees C. Two dominant modes of failure have been identified: firstly, fusion boundary failure at high stresses; and, secondly, Type IV failure at low stresses. The stress level at which the switch in failure mechanism takes place has been found to be associated with the relative creep resistance and lifetimes, over a wide range of uniaxial stresses, for parent, heat affected zone, Type IV and weld materials. The equi-biaxial stress loading condition (mean diameter stress equal to the axial stress) has been confirmed to be the worst loading condition. For this condition, simple design formulae are proposed for both 590 and 620 degrees C. PMID- 16243709 TI - Opposing actions of STAT-1 and STAT-3. AB - The signal transducers and activators of transcription (STATs) are a family of transcription factors, which were originally identified on the basis of their ability to transduce a signal from a cellular receptor into the nucleus and modulate the transcription of specific genes. Interestingly, recent studies have demonstrated that STAT-1 plays a key role in promoting apoptosis in a variety of cell types, whereas STAT-3 has an anti-apoptotic effect. Moreover, whilst STAT-3 promotes cellular proliferation and is activated in a variety of tumour cells, STAT-1 appears to have an anti-proliferative effect. Although the initially characterised signal transduction events mediated by STAT-1 and STAT-3 involve the DNA binding and transcriptional activation domains of the factor, some of their other effects appear not to require DNA binding. Therefore, STAT-1 and STAT 3 can mediate the regulation of gene transcription both by direct DNA binding and via a co-activator mechanism and despite their very similar structures, have antagonistic effects on cellular proliferation and apoptosis. PMID- 16243710 TI - Mdm2 in growth signaling and cancer. AB - Genetic and biochemical evidence have demonstrated a direct link between Mdm2 and cancer development. Elevated expression of Mdm2 is observed in a significant proportion of different types of cancer. The major contribution of Mdm2 to the development of cancer is through a tight inhibition of the activities and stability of the tumor suppressor p53. However, extensive studies over the past few years have identified p53-independent functions of Mdm2, in the regulation of several important cellular processes and multiple signaling pathways. The promotion of cell cycle progression by Mdm2 is mediated via p53 inhibition, and by regulating the pRb/E2F complex. Mdm2 is an important mediator of growth and survival signaling in the PI3K/Akt pathway, an activator of certain steroid hormone receptors, and an inhibitor of the TGF-beta growth restrictive pathway. Thus, the impact on these pathways by deregulated Mdm2, as often observed in cancer, can be oncogenic in a permissible environment. This renders Mdm2 as an important target for the development of anti-cancer drugs. PMID- 16243711 TI - Adapter protein connections: the MRL and Grb7 protein families. PMID- 16243712 TI - The priming/completion paradigm to explain growth factor-dependent cell cycle progression. AB - Approximately 50 years ago, researchers established conditions to maintain cells in tissue culture: Likely et al. (1952), Scherer et al. (1953), Eagle (1955). This simple model system set the stage for discovery of growth factors and the signaling systems that they engage to mediate cellular responses such as proliferation. The purpose of this review is to present the original view of how growth factors regulate cell cycle progression and an updated (priming/completion) version of how growth factors advance resting cells through the cell cycle. PMID- 16243713 TI - The conserved lymphokine element 0 is a powerful activator and target for corticosteroid inhibition in human interleukin-5 transcription. AB - The role of eosinophilia in allergic disorders indicates hIL-5 as a potential target for therapy. The conservation of hIL-5 gene proximal elements suggests they are important in controlling expression. Corticosteroids are important in the treatment of allergy, and are powerful inhibitors of IL-5 expression. This study aimed at understanding the role of hIL-5 conserved proximal elements, and elucidating the target of corticosteroid activity, in hIL-5 gene expression. Methods used include transient transfection of PBMC and PER117 cells with hIL-5 deletion constructs, EMSA, Western Blotting, and RT-PCR. The conserved proximal CLE0/TATA elements driving a reporter gene gave similar or higher expression than a 500 bp promoter in primary human T cells and a T-cell line. Two and three copies of IL-5 CLE0 upstream of the silent IL-4 minimal promoter gave 30-45 fold increases in expression in forward orientation, but little activity in reverse orientation. Consequently, CLE0 is a powerful activator but not a classical enhancer. Deletion analysis identified CLE0 as the key element in the inhibition of IL-5 reporter constructs by dexamethasone, and RT-PCR analysis indicated that GILZ expression correlated with dexamethasone-induced inhibition of IL-5. Ectopic expression of GILZ, confirmed by western blotting, gave a 90% inhibition of promoter constructs in absence of dexamethasone. CLE0 is a powerful activator sufficient for the inducible expression of IL-5, and functions when moved upstream in a heterologous promoter. CLE0 is also the main target for IL-5 inhibition by dexamethasone, and we present evidence consistent with a role of GILZ in this. PMID- 16243714 TI - Identification of growth factor-regulated proteins using 2D electrophoresis and mass spectrometry. AB - Proteomic technology has recently emerged as a powerful tool for detecting both qualitative and quantitative changes of proteins that occur upon activation of complex signaling pathways. In the present study, comparison of the protein profile of platelet-derived growth factor (PDGF), epidermal growth factor (EGF) and nerve growth factor (NGF)-stimulated and unstimulated cells with two dimensional electrophoresis followed by mass spectrometric analysis led to the identification of a number of proteins, several of which had not been previously shown to be regulated by receptor-tyrosine kinases. Using subcellular fractionation, our approach was able to identify not only changes due to altered gene transcription, but also due to intracellular protein translocation or modification. One of the proteins that was identified among other PDGF-regulated molecules was prohibitin, a potential tumour suppressor previously implicated in cell cycle regulation and protection of mitochondrial proteins from degradation. Further analysis confirmed that mitochondria-associated prohibitin translocates to an insoluble perinuclear compartment. This study demonstrates the utility of proteomic strategies in identifying potential growth factor-regulated effectors. PMID- 16243715 TI - C-terminal Src kinase (CSK) and CSK-homologous kinase (CHK)--endogenous negative regulators of Src-family protein kinases. AB - C-terminal Src kinase (CSK) and CSK-homologous kinase (CHK) are endogenous inhibitors of the Src-family protein tyrosine kinases (SFKs). Since constitutive activation of SFKs contributes to cancer formation and progression, to prevent excessive activation of SFKs, their activity in normal cells is kept at the basal level by CSK and CHK. CSK and CHK inactivate SFKs by specifically phosphorylating a consensus tyrosine (called Y(T)) near their C-termini. Upon phosphorylation, the phospho-Y(T) engages in intramolecular interactions that lock the SFK molecule in an inactive conformation. SFKs are anchored to the plasma membrane, while CSK and CHK are localized predominantly in the cytosol. To inhibit SFKs, CSK and CHK need to translocate to the plasma membrane. Recruitment of CSK and CHK to the plasma membrane is mediated by the binding of their SH2, SH3 and/or kinase domains to specific transmembrane proteins, G-proteins and adaptor proteins located near the plasma membrane. For CSK, membrane recruitment often accompanies activation. CSK and CHK employ two types of direct interactions with SFKs to achieve efficient Y(T) phosphorylation: (i) short-range interactions involving binding of the active sites of CSK and CHK to specific residues near Y(T), (ii) long-range non-catalytic interactions involving binding of SFKs to motifs located distally from the active sites of CSK and CHK. The interactions between CSK and SFKs are transient in nature. Unlike CSK, CHK binds tightly to SFKs to form stable protein complexes. The binding is non-catalytic as it is independent of Y(T). More importantly, the tight binding alone is sufficient to completely inhibit SFKs. This non-catalytic inhibitory binding represents a novel mechanism employed by CHK to inhibit SFKs. Given that SFKs are implicated in cancer development, compounds mimicking the non-catalytic inhibitory mechanism of CHK are potential anti-cancer therapeutics. PMID- 16243716 TI - Enzyme therapy for management of coeliac disease. AB - OBJECTIVE: Enzyme therapy based on animal digestive extracts was investigated as a means of completely digesting toxic residues from gluten in the small intestine, thus providing a means of protection of the mucosa. MATERIAL AND METHODS: A randomized, placebo-controlled, clinical trial of an encapsulated enzyme extract was conducted in 21 coeliac patients in remission who were challenged with a modest amount of gluten daily over 2 weeks. Enzyme extract (900 mg) in three divided doses was administered during this challenge to half the group and a placebo to the other half in a double-blind, crossover design. Symptoms were recorded in daily diaries; blood was taken for tissue transglutaminase antibodies (anti-tTG) at the start and at intervals up to 12 weeks. Duodenal biopsies were performed for histological assessment at the start and end of each challenge period for 6 patients chosen at random from volunteers. After a further 10 weeks, the groups were changed over, and the same assessments carried out. RESULTS: Only 8 of the 21 patients (38%) had more than 5 episodes of moderate to severe symptoms during either of the gluten challenge periods, and in these, symptoms scores were ameliorated during enzyme therapy compared with the placebo period (p<0.02). Rises of 5 U/ml or more in anti-tTG occurred in only 5 patients at about 6-8 weeks after challenge, but were not correlated with symptoms. CONCLUSIONS: Only 1 of the 6 patients had normal histology at entry, thus focusing attention on the need for better management of the disease. By histological criteria, enzyme therapy offered better protection than placebo during the gluten challenges. The study supports the use of enzyme supplementation as a safeguard for patients with coeliac disease because of the difficulty of ensuring a strictly gluten-free diet. PMID- 16243718 TI - Adducts between nucleophilic amino acids and hexahydrophthalic anhydride, a structure inducing both types I and IV allergy. AB - Haptens causing type I allergy have been shown to predominantly form lysine adducts in the carrier protein, while many haptens giving rise to type IV allergy preferentially form adducts with cysteine residues. Hexahydrophthalic anhydride derivatives are strong sensitizers capable of inducing allergic rhinitis, asthma and urticaria (type I allergy) and allergic contact dermatitis (type IV allergy). The ability of hexahydrophthalic anhydride (HHPA) to form adducts with nucleophilic amino acids and a model peptide in vitro is presented. Adduct formation was monitored by high-performance liquid chromatography with ultraviolet light/vis detection (LC-UV/vis) and high-performance liquid chromatography with mass spectrometric detection (LC/MS). The characterization was obtained by nuclear magnetic resonance spectroscopy (NMR) and mass spectrometry (MS and MS/MS). It was found that HHPA formed adducts with N(alpha) acetylated lysine and cysteine and the non-acetylated alpha-amino group of proline and, to some extent, also with other nucleophilic amino acids. The adducts with lysine and proline were chemically stable. Addition of one HHPA to a model carrier peptide with all important nucleophilic amino acid residues showed N-terminal proline to be the major site of reaction. The addition of a second hapten gave a lysine adduct, but a minor cysteine adduct was also found. The cysteine-HHPA adducts were shown to be chemically unstable and participated in further reactions with lysine forming lysine-HHPA adducts. The results will be useful for understanding the formation of HHPA-protein adducts with the capability of being markers of exposure, and also to a deeper understanding of the chemical structures causing types I and IV allergy. PMID- 16243717 TI - Prevalence and clinical significance of sonographic detection of enlarged regional lymph nodes in Crohn's disease. AB - OBJECTIVE: Reactive regional lymph node enlargement is a frequent ultrasonographic finding in patients with Crohn's disease. However, the prevalence of this condition and its clinical significance are unknown. This study assesses the prevalence of enlarged regional mesenteric lymph nodes and its clinical significance in Crohn's disease, and in particular whether there is a correlation between the sonographic detection of enlarged regional lymph nodes and the degree of clinical or biochemical activity of the disease. MATERIAL AND METHODS: A total of 240 in- and outpatients with Crohn's disease underwent intestinal ultrasound to assess the presence of enlarged regional lymph nodes as well as the thickness and echopattern of the bowel wall, the site and extent of Crohn's disease and the presence of stenosis, fistulas and abscesses. Demographic, clinical and biochemical parameters were also collected for each patient. A multivariate model by means of multiple regression analysis was used to identify independent variables linked to regional lymph node enlargement. RESULTS: Enlarged regional lymph nodes were detected ultrasonographically in 25.4% of Crohn's disease patients. The presence of regional lymph nodes showed a weak correlation with both clinical and biochemical Crohn's disease activity. Regional lymph nodes were found more frequently in young patients (50% of patients < 30 years, 18% of patients between 30 and 50 yrs, and 7% of patients > 50 yrs; p<0.0001) and in patients with a shorter disease duration. Enlarged regional lymph nodes were strongly correlated with internal fistulas and intra abdominal abscesses. The multiple regression analysis showed that age, duration of disease and presence of internal fistulas were the best independent predictive factors linked to the presence of enlarged mesenteric lymph nodes. CONCLUSIONS: The sonographic detection of enlarged regional lymph nodes is more frequent in young patients, which suggests an earlier phase of Crohn's disease and the presence of septic complications such as fistulas and abscesses, but this is of limited valued in assessing disease activity. PMID- 16243719 TI - Biological monitoring among benzene-exposed workers in Bangalore city, India. AB - Environmental and biological monitoring was carried out in the winter season of 2004 for 30 gasoline station workers (study subjects) and 30 office workers (controls) of Bangalore city, India. Personal air sampling was carried out in the breathing zone of workers using an Anasorb CSC sorbent tube (SKC 226-01) fitted to the low-flow personal samplers (PCXR4 and pocket pump Model No. 210-1002) at a flow rate of 200 ml min(-1) during the shift work of 8 h. The benzene content adsorbed in the sorbent tube (SKC 226-01) was desorbed with 1 ml of benzene-free carbon disulfide on a developing vibrator and later analysed by Trace GC fitted with MXT-624 column and flame ionization detector. The mean time weighted average benzene concentration found among study and controls was 1.10+/-1.08 and 0.070+/ 0.035 mg m(-3), respectively. Biological monitoring for benzene exposure was performed by measuring trans,trans muconic acid (t,t-MA) in the end shift urine samples using HPLC-UV technique. End-shift urine samples (1 ml) were adjusted to pH 7-9 with phosphate buffer pH 7.4 passed through the preconditioned Q-SAX anion exchange cartridge and the (t,t-MA) is extracted with 10% acetic acid and later analysed by HPLC-UV detection The mean t,t-MA found among study and controls were 563.16+/-281.81 and 266.88+/-110.65 microg g(-1) creatinine. About 50% of the study subjects (15) have higher t,t-MA values than the biological exposure index of the American Conference of Government Industrial Hygienist (ACGIH). Correlation is significant at 5% level (p<0.05) between personal air benzene concentration and urinary t,t-MA in the study group. Based on these findings, the t,t-MA can be used as a biomarker for benzene exposure. PMID- 16243720 TI - Application of two SH-based methods for metallothionein determination in mussels and intercalibration of the spectrophotometric method: laboratory and field studies in the Mediterranean Sea. AB - Metallothionein (MT) induction is widely used as a biomarker of exposure to metals in mussels. The aims of the present work were first to compare the suitability of spectrophotometry and differential pulse polarography (DPP) for MT detection in mussels exposed to 200 ppb cadmium for 9 days in a laboratory experiment and in mussels sampled in different seasons from expected pollution gradients along the Mediterranean Sea; second, to intercalibrate the widely used spectrophotometric method using mussels from Saronikos Gulf. In the intercalibration of the spectrophotometric method, similar results (p>0.05) were obtained by two different research teams indicating a good reproducibility of the technique. However, polarographic and spectrophotometric methods gave significantly (p<0.05) different results in laboratory and field studies. In the laboratory experiment, MT values detected with DPP were nine times higher than with spectrophotometry. The results obtained by the two methods were significantly correlated. Both methods could discriminate between control and exposed mussels. In field studies, MT values obtained by DPP were 34-38-fold higher than with spectrophotometry, and MT concentrations measured by both methods were not correlated. This discrepancy could be due to several factors, including the low levels of bioavailable metals in the studied areas and the possibility that the different methods can measure MT isoforms differentially. Further work is needed to decipher the functions of MT isoforms in mussels. This information is relevant for the application of MT as a biomarker in biomonitoring programmes. PMID- 16243721 TI - Do metals inhibit acetylcholinesterase (AChE)? Implementation of assay conditions for the use of AChE activity as a biomarker of metal toxicity. AB - The enzymatic activity of acetylcholinesterase (AChE) has been shown to be altered by environmental contaminants such as metals. However, the available literature illustrates a background of contradictory results regarding these effects. Therefore, the main purpose of this study was to investigate the potential of five metal ions (nickel, copper, zinc, cadmium and mercury) to inhibit AChE activity in vitro. First, to accomplish this objective, the possible interference of metals as test toxicants in the Ellman's assay, which is widely used to assess AChE activity, was studied. The potential influence of two different reaction buffers (phosphate and Tris) was also determined. The results suggest that the selected metals react with the products of this photometric technique. It is impossible to ascertain the artefactual contribution of the interaction of the metals with the technique when measuring AChE inhibition. This constitutes a major obstacle in obtaining accurate data. The presence of phosphate ions also makes enzymatic inhibition difficult to analyse. Attending to this evidence, an assay using the substrate o-nitrophenyl acetate and Tris buffer was used to investigate the effects of metals on AChE activity. O-nitrophenyl acetate is also a substrate for esterases other than cholinesterases. It is therefore only possible to use it for the measurement of cholinesterase activity with purified enzymes or after a previous verification of the absence of other esterases in the sample tissue. Under these conditions, the results indicate that with the exception of nickel, all tested metals significantly inhibit AChE activity. PMID- 16243722 TI - Prenatal exposure to pesticides: analysis of human placental acetylcholinesterase, glutathione S-transferase and catalase as biomarkers of effect. AB - Pre- and perinatal exposure to pesticides is deleterious on foetal and neonatal development, but information regarding possible effects on environmental low-dose exposure to pesticides is scarce. Most epidemiological studies of the health effect of pesticides have been based on self-reported information. However, detailed information on past pesticide use is difficult to reconstruct. This is a current study conducted among pregnant mothers attending a delivery care and perinatal programme at a public hospital. The study investigates biomarkers of early effects in placentas from women living in an area with an intensive use of pesticides in the northern part of Patagonia, province of Rio Negro, Argentina, and it assesses the consistency of the information provided by self-reports. The study confirms that placental acetylcholinesterase and catalase activities are significantly associated with periods of organophosphorus pesticides application, while glutathione S-transferase is not affected. We found a positive correlation between environmental exposure to organophosphorus pesticides and carbamate insecticides and newborn head circumference. The findings provide a further indication of a link between placenta acetylcholinesterase and catalase activity and prenatal exposure to pesticides in population studies. Both placenta enzymes may be used as biomarkers in health surveillance programmes for early diagnosis of exposure related alterations produced by organophosphorus pesticides and carbamate pesticides. PMID- 16243723 TI - Preliminary studies on the effect of moderate physical activity on blood levels of glutathione. AB - Molecular epidemiological approaches are being used to study how physical activity may protect against cancer. Prior epidemiological data suggest that physical activity protects against lung cancer; however, interpretation of these data is complicated by potential confounding by smoking. Glutathione (GSH) detoxifies cigarette smoke carcinogens and the paper tests whether physical activity levels are associated with blood GSH levels. Study subjects were enrolled in a chemoprevention trial testing whether antioxidant micronutrient supplementation reduces genetic damage from cigarette smoking. Physical activity data were collected by questionnaire from 178 subjects at 12 months of follow-up in the trial. Total GSH (tGSH), which is the sum of free and protein-bound GSH and glutathione disulfide levels, was measured using the 5,5'-dithiobis-(2 nitrobenzenoic acid) colormetric assay with red blood cell samples collected at the 12-month time point. In multivariate linear regression analyses that controlled for gender and cigarettes smoked per day, tGSH was positively associated with hours per week of moderate intensity activity (beta=0.005, p=0.02). Hours per week of vigorous intensity activity were unassociated with tGSH and the effect of moderate activity remained after control for vigorous activity. The results are consistent with prior research showing differential effects of moderate and vigorous activity and suggest a mechanism through which physical activity may influence lung cancer risk. PMID- 16243724 TI - Expression of survivin mRNA in dog tumors. AB - Survivin, a member of the inhibitor of apoptosis (IAP) gene family, overexpresses in various human tumors. Recently this protein has attracted strong interest as a potential prognostic marker because it promotes malignancy through anti-apoptotic activity and is associated with a more aggressive phenotype. To explore the utility of survivin as a veterinary marker of tumor malignancy, we performed molecular cloning of dog survivin cDNA and studied survivin mRNA expression in a variety of naturally occurring dog tumors. The dog cDNA contains a 426-bp open reading frame encoding 142 amino acids of polypeptide, in which a structure termed the baculovirus IAP repeat (BIR) domain, commonly observed in IAPs, is found, as it is in other mammalian survivin protein. The transcript was detected in many adult normal organs including heart, lung, liver, stomach, duodenum, colon, spleen, kidney and testis. As a result of quantitative expression analysis by real-time PCR undertaken for benign and malignant tumors, overexpression of the survivin gene was found in 3 of 18 malignant tumors and in none of the benign tumors, suggesting that survivin overexpression is associated with tumor malignancy in dog. PMID- 16243725 TI - Cloning and expression analysis of a vacuolar Na+/H+ antiporter gene from alfalfa. AB - A full-length cDNA, named MsNHX1, encoding a vacuolar Na+/H+ antiporter was cloned from alfalfa (Medicago sativa), using degenerate primers, followed by 3' and 5' RACE. The cDNA sequence was 2225 bp long and included an open reading frame encoding a deduced protein of 541-amino-acid polypeptide. The deduced amino acid sequence showed high similarity (more than 73%) to those of the previously cloned Na+/H+ antiporters form Arabidopsis thaliana, Qryza sativa, Atriplex gemlinin, Beta vulgaris and Hordeum vulgare. Southern hybridization showed that MsNHX1 has only one copy in alfalfa genome. Semiquantitative RT-PCR analysis indicated that the mRNA level of MsNHX1 was moderate without stress and markedly up-regulated after treatment by NaCl and ABA (abscisic acid). Those results suggest that the MsNHX1 product play an important role in salt tolerance of the alfalfa, and its transcript expression is possibly partially regulated through ABA-dependent signaling pathway. PMID- 16243726 TI - Isolation and characterization of TaDof1 transcription factor in wheat (Triticum. aestivum. L). AB - The Dof (DNA binding with one finger) proteins are plant specific transcription factors. Dof proteins are apparently encoded by a multiple gene family in higher plants. However, only one Dof gene, WPBF, was reported in wheat. In this study, a member of Dof gene family, TaDof1, was cloned from wheat. TaDof1 encode 291 amino acids, with a predicted molecular mass of 30.348 kDa. At its N-terminal end, a 52 amino acid stretch typical of Dof domain and two serine-rich stretches were observed. Sequence alignment indicated that, in Dof domain, TaDof1 share more than 75% identity with other Dof proteins of different species. TaDof1 was expressed highly in leaves and sheaths, but lowly in roots, and constitutively expressed in developing seeds of 2-12 DAP. It was interesting to note that TaDof1 was differentially expressed between hybrids F1 and parents in root, sheath and leaf. The implication of the differential expression patterns of TaDof1 was discussed in related to the up-regulation of C4 pathway related gene in hybrid rice and heterosis. PMID- 16243727 TI - Cloning, sequencing and salt induced expression of PEAMT and BADH in oilseed rape (Brassica napus). AB - Agriculture productivity is severely hampered by soil salinity, drought and other environmental stresses. Studies on stress-resistant plants (halophytes, xerophytes, accumulating plants for specific toxic ions) have illuminated some mechanisms of stress tolerance in plants at metabolic or molecular levels, which gave some clues on how to genetically engineer stress-tolerant crops. With the isolation of more stress-responsive genes, genetic engineering with modified expression of stress responsive genes may be an effective way to produce stress tolerant crops. In the present report, two genes (PEAMT and BADH) encoding the corresponding key enzymes for choline and glycine betaine (an important osmoprotectant) biosynthesis in plants were isolated in oilseed rape, an important oil crop in the world. Effects of salt stress on their expression were studied with quantitative PCR and their potential use in the genetic engineering of oilseed rape was discussed. PMID- 16243728 TI - Structural analysis of a peptide synthetase gene required for ergopeptine production in the endophytic fungus Neotyphodium lolii. AB - Lysergyl peptide synthetase 1 catalyzes the assembly of toxic ergopeptines from activated D-lysergic acid and three amino acids. The gene encoding this enzyme in the endophytic fungus Neotyphodium lolii was analyzed and compared to a homologous gene from the ergot fungus Claviceps purpurea. Each gene contained two introns, which were found in the same relative position within two modules of the gene. The 5' ends of the two genes were unusually divergent. Signature sequences determining substrate specificity were similar in adenylation domains that recognized identical amino acids but differed within the adenylation domain for the amino acid that varies between the major ergopeptines of the two fungi. Homologues were detected in several related endophytic fungi; the tall fescue endophyte Neotyphodium coenophialum contained a divergent, second copy of the gene. Our results provide new information on the structure and distribution of this important peptide synthetase involved in ergot alkaloid biosynthesis. PMID- 16243729 TI - Cloning and characterization a novel human 1-acyl-sn-glycerol-3-phosphate acyltransferase gene AGPAT7. AB - The 1-Acylglycerolphosphate acyltransferase is crucial enzyme for synthesis of glycerolipids as well as triacylglylcerol biosynthesis in eukaryotes. Six members of 1-acyl-sn-glycerol-3-phosphate acyltransferase family in human have been described, which were AGPAT1, 2, 3, 4, 5 and 6. Here we report the cloning and characterization of another novel human 1-acyl-sn-glycerol-3-phosphate acyltransferase member AGPAT7 (1-acyl-sn-glycerol-3-phosphate acyltransferase 7) gene, which was mapped to human chromosome 15q14. The AGPAT7 cDNA is 1898 bp in length, encoding a putative protein with 524 amino acid residues, which contains an acyltransferase domain in 123-234 aa. RT PCR amplification in 18 human tissues indicated that human AGPAT7 gene was widely expressed in uterus, thymus, pancreas, skeletal muscle, bladder, stomach, lung and testis. AGPAT7 protein was mainly localized to the endoplasmic reticulum (ER) in Hela cells. PMID- 16243731 TI - Cerebral embolic protection devices and percutaneous carotid artery stenting. PMID- 16243732 TI - Acute and intermediate-term procedural results using cypher stenting to treat multi-vessel coronary artery disease. AB - OBJECTIVE: We examined our experience using the sirolimus eluting stents (Cypher) as an alternative to surgical revascularization in carefully selected cohort of patients undergoing multi-vessel percutaneous coronary intervention. METHODS: Fifty consecutive patients with multi-vessel disease who were good candidates for both surgical and percutaneous revascularization were included in the current analysis. All patients underwent a careful clinical evaluation prior to the intervention, and they were followed for procedural and clinical outcomes for nine months. RESULTS: Mean age was 64+/-11 years (40 males, 30% diabetics) and 10 patients (20%) had three-vessel disease. Angina class was 2.7+/-0.6 at baseline. Overall, 116 lesions were treated using 122 stents (mean 2.4 stents per patient). Total mean stent length was 43+/-12 mm (range: 21-90 mm). Overall, one patient died during follow-up (2%), no patient had stroke or Q wave MI and one patient experienced non-Q myocardial infarction. There was no documented stent thrombosis and two patients (4%) underwent target-vessel revascularization. The hierarchical cumulative major adverse cardiac event rate was 8% and the cardiac event-free survival rate was thus 92%. CONCLUSION: Multi-vessel stenting using Cypher stents is a viable treatment strategy in selected group of patients with multi-vessel coronary artery disease. It is associated with excellent intermediate-term clinical outcomes and thus it could serve as the primary revascularization strategy of choice in appropriate candidates. PMID- 16243733 TI - Carotid artery stenting: update. AB - "Extracranial carotid artery stenosis is responsible for approximately 20-30% of ischemic strokes. Traditionally, carotid artery stenosis has been treated with carotid endarterectomy. However, the low periprocedural complication rate and the mid term durability of carotid artery stenting has made it a competitive alternative treatment strategy. This update reviews the technical aspects of carotid artery stenting, clinical data supporting carotid artery stenting particularly in high risk patients, and the complications associated with carotid artery stenting." PMID- 16243734 TI - Intracardiac echocardiography guidance during percutaneous transluminal septal myocardial ablation in patients with obstructive hypertrophic cardiomyopathy. AB - BACKGROUND: Percutaneous transluminal septal myocardial ablation (PTSMA) recently emerged as an alternative to myectomy for hypertrophic obstructive cardiomyopathy (HOCM) patients with drug-refractory symptoms. The target septal branch selection is a main point to achieve the therapeutic result. METHODS AND RESULTS: We report about PTSMA performed using intracardiac echocardiography (ICE) to guide the procedure in 9 symptomatic HOCM patients. The target septal branch was chosen on the basis of the risk-area visualized using ICE after injection of a contrast agent. During alcohol administration a backscattered signal enhancement of the infarcted area was detected. The procedures were uncomplicated and effective to reduce the gradient from 78.9+/-20.4 mmHg to 7.8+/-7.9 mmHg (p<0.0001). CONCLUSIONS: In this initial experience ICE monitoring during PTSMA was safe and provided high quality and continuous imaging of the treated segment of the septum during the whole procedure. PMID- 16243735 TI - Intracardiac echocardiographic guidance for hemodynamic assessment in a patient with congenital abnormalities and a prosthetic aortic valve. AB - In this case report, we present the use of intracardiac echocardiography (ICE) for guiding the cardiac catheterization and subsequent hemodynamic investigation in an unusual patient case with multiple congenital abnormalities (bicuspid aortic valve, left cervical aortic arch, two aortic coarctations) and two aortic valve replacement operations in the past. The ICE catheter (AcuNav) permitted us to accurately and safely puncture the interatrial septum and place the Swan-Ganz catheter in the left ventricle; additionally, visualization of the aortic coarctation in the ascending aorta was also achieved. PMID- 16243736 TI - Three-dimensional coronary reconstruction from routine single-plane coronary angiograms: in vivo quantitative validation. AB - BACKGROUND: Current X-ray technology displays the complex 3-dimensional (3-D) geometry of the coronary arterial tree as 2-dimensional (2-D) images. To overcome this limitation, an algorithm was developed for the reconstruction of the 3-D pathway of the coronary arterial tree using routine single-plane 2-D angiographic imaging. This method provides information in real-time and is suitable for routine use in the cardiovascular catheterization laboratory. OBJECTIVES: The purpose of this study was to evaluate the precision of this algorithm and to compare it with 2-D quantitative coronary angiography (QCA) system. METHODS: Thirty-eight angiographic images were acquired from 11 randomly selected patients with coronary artery disease undergoing diagnostic cardiac catheterization. The 2 D images were analyzed using QCA software. For the 3-D reconstruction, an algorithm integrating information from at least two single-plane angiographic images taken from different angles was formulated. RESULTS: 3-D acquisition was feasible in all patients and in all selected angiographic frames. Comparison between pairs of values yielded greater precision of the 3-D than the 2-D measurements of the minimal lesion diameter (P<0.005), minimal lesion area (P<0.05) and lesion length (P<0.01). CONCLUSIONS: The study validates the 3-D reconstruction algorithm, which may provide new insights into vessel morphology in 3-D space. This method is a promising clinical tool, making it possible for cardiologists to appreciate the complex curvilinear structure of the coronary arterial tree and to quantify atherosclerotic lesions more precisely. PMID- 16243737 TI - Functional T-lymphocyte dichotomy in the peripheral blood of patients with unstable angina. AB - OBJECTIVES: Herein, we investigated the percentage of T-helper (Th1) and Th2 cells among the general T-cell population in the peripheral blood of patients with stable angina (SA) and unstable angina (UA). BACKGROUND: Recent evidence suggests that Th1 cells and the cytokines that they secrete (especially IFN gamma) have a role in the activation of macrophages, promotion of clot formation and destabilization of atherosclerotic plaques. Thus, Th1 cytokines may contribute to the initiation and progression of UA. In contrast, cytokines secreted by Th2 cells (e.g. IL-10) are known to inhibit activation and proliferation of Th1 cells and the secretion of IFN-gamma, lysosomal enzymes and metalloproteinases. Therefore, we sought to examine whether the ratio of IFN gamma to IL-10 secreting cells is altered in patients with UA. METHODS: The percentage of Th1 and Th2 cells among the general T-cell population was determined by fluorescent intracellular cytokine staining (IFN-gamma and IL-10, out of the total CD3 positive cells). RESULTS: The percentage of T-cells positive for intracellular IFN-gamma was significantly higher in patients with UA (n = 22) in comparison with SA (n = 20) patients (39.0+/-2.8% and 29.6+/-2.7%, respectively. P = 0.02). There was no significant difference in intracellular IL 10 positive cells between the two groups. In addition, there was no significant difference in the ratio between the intracellular IFN-gamma positive cells and the intracellular IL-10 positive cells. CONCLUSIONS: There is an increased activity of Th1 cells in patients with UA in comparison with patients with SA. There is no evidence of heightened activity of Th2 cells in either group. Thus, IFN-gamma secreted by peripheral blood T-lymphocytes,may be an important immunomodulator contributing to destabilization of the atheromatous plaque lying at the base of the etiopathogenesis of unstable angina. PMID- 16243738 TI - CT angiography is here--are we expected to see a change of angiography referral pattern? AB - A 52-year-old asymptomatic man, with cardiac risk factors of hypertension, Type II diabetes, hypertriglyceridemia, low HDL, obesity, and positive family history for early coronary artery disease (CAD), was referred to nuclear stress test. He exercised for 14 minutes, achieved his target heart rate, without any symptoms or ECG changes. Nuclear images were entirely normal. However, the patient was still concerned and anxious so he underwent CT angiography that revealed coronary narrowings. Next, he underwent coronary angiography with similar findings. The lesions were treated with stents. We present a case report to illustrate how newer technology such as CT angiography alters the way in which we diagnose CAD and decide on whether to pursue further invasive therapy. PMID- 16243739 TI - Seeing transient regional myocardial ischaemia through new eyes. PMID- 16243740 TI - Middle ear pathology in ancient and modern populations: incudal osteoma. AB - CONCLUSIONS: Osteomas of the incus were found in surprisingly high numbers in ancient and modern populations. The coexistence of osteoma formation and unique vascularization on the medial surface of the incus alludes to a relationship between the two findings. OBJECTIVE: Osteomas involving the middle ear ossicles are rare. The aim of this study was to compare the incudal pathology of ancient and recent periods using the recent literature. MATERIAL AND METHODS: Pathologies encountered in 1170 human incudes belonging to different populations from ancient and recent periods were studied. Most of the ossicles were found in the middle ear of dry skulls or during dissection. The observations were done with a binocular microscope and a Nikon Profile Projector. RESULTS: A total of 47 incudes (4.01%) presented with a variety of pathologies, osteomas being found in 19 cases (1.62%). The osteomas were noted only on the medial surface of the incus. Other findings, including prominent erosions, multiple nutrient foramina and pitting, were also found on the medial side of the incus. The incidence of osteoma was found to be more common in ancient populations (1.88%) and most prevalent among Bedouins (4.55%). PMID- 16243741 TI - Tonic contractions of the tensor tympani muscle: a key to some non-specific middle ear symptoms? Hypothesis and data from temporal bone experiments. AB - CONCLUSIONS: The results of this study show that in clinical practice it will not be easy to diagnose tonic contractions of the tensor tympani muscle and only a combination of findings will be helpful. Based on these experimental results a clinical study will be started which should clarify the diagnostic relevance of indicators of tonic tensor muscle contractions. OBJECTIVES: There are indications from the literature and from personal experience that tonic contractions of the tensor tympani muscle may play a role in some ear symptoms, such as fullness, certain cases of tinnitus, slight hearing loss or Meniere's disease-like findings. In order to prove this theory we looked for indicators, either visual or functional, to help clinically diagnose the functional state of the muscle, particularly its tonic contraction. MATERIAL AND METHODS: Experiments simulating tensor contractions were carried out on temporal bone specimens. Traction was applied either to the isolated muscle, to its tendon or to the malleus neck. Effects were observed either visually via an endoscope or by impedance audiometry using multiple-frequency tympanometry. RESULTS: During simulated tensor traction the aspect of the tympanic membrane changed slightly, i.e. there was some inward movement of the umbo. However, such effects were only identifiable during the pulling action or by directly comparing the "contracted" and "relaxed" states. Tympanometry revealed a decrease in the peak amplitudes and a shift in the middle ear resonance towards higher frequencies during contractions. PMID- 16243742 TI - Selective regeneration of motor and sensory axons in an experimental peripheral nerve model without endorgans. AB - We assessed the selectivity of motor and sensory axon regeneration towards the distal motor and sensory nerve segments that were disconnected from endorgans in a rat silicone Y chamber model. The L5 ventral root was used as a pure motor nerve, and the saphenous nerve was used as a sensory nerve. In experiment 1 (n=11), the proximal stump of the L5 ventral root, a 1-cm-long L5 ventral root segment and a saphenous nerve segment were inserted into a silicone Y chamber. In experiment 2 (n=11), the proximal stump of the saphenous nerve, a L5 ventral root segment and a saphenous nerve segment were inserted into a Y chamber. The distance between the nerve stumps was 5 mm. Six weeks later, the number of regenerated myelinated motor and sensory axons was measured and compared in the distal two channels. Motor axons showed no selective regeneration, but sensory axons did. PMID- 16243743 TI - Clinical reappraisal of the segmental pectoralis major turn-over flap for coverage of the local mediastinal wound. AB - The viability of a long and narrow pectoralis major turn-over flap based on one intercostal muscle perforator was supported by the result of our cadaveric injection study. This flap preserves the muscular strength and aesthetic contour of the anterior axillary line, and is attractive for reconstruction of the mid sternum. PMID- 16243744 TI - Sensorial donor site morbidity after saphenous neurocutaneous flap. AB - Neurocutaneous flaps based on the arterial network around the superficial sensory nerves are popular in soft tissue coverage of the lower extremities, and are usually preferred in reconstruction of the lower leg and foot. Although sacrifice of sensory cutaneous nerves is mentioned as one of their major disadvantages to our knowledge the morbidity in the foot and at the donor site has not been properly investigated. We investigated sensorial morbidity in the foot and at the donor site caused by raising a saphenous neurocutaneous flap in 14 patients by using static two-point discrimination test, Semmes-Weinstein monofilament test, vibration test, and by measuring somatosensory evoked potentials in the twelfth postoperative month. Our results suggest that sensory recovery is good and protective sensation is gained in most patients. PMID- 16243746 TI - Cognitive appraisal accuracy moderates the relationship between injury severity and psychosocial outcomes in traumatic brain injury. AB - OBJECTIVE: Traumatic brain injury (TBI) frequently results in significant changes in physical, cognitive and emotional status. Outcomes after TBI may be related to accurate appraisal of these changes. This study examined the relationship between cognitive appraisal accuracy and psychosocial outcomes in TBI survivors. METHODS: Participants were male and female TBI survivors (n=103) who were at least 6 months post-injury and a family member of each survivor. Appraisal accuracy was assessed using self- and observer-report measures of perceived cognitive difficulties. Family members also completed a measure of psychosocial function. Hierarchical regression techniques were used to determine whether cognitive appraisal accuracy accounted for a significant proportion of variance in psychosocial outcomes. RESULTS: Study findings indicated cognitive appraisal accuracy moderated the relationship between injury severity and aspects of psychosocial function. CONCLUSIONS: The results suggest that brain injury outcomes may be improved when an individual is able to accurately assess limitations. PMID- 16243747 TI - Exploring the impact of traumatic brain injury on the older couple: 'yes, but how much of it is age, I can't tell you ...'. AB - PRIMARY OBJECTIVE: To qualitatively explore the partner relationships of older couples faced with traumatic brain injury (TBI). RESEARCH DESIGN: Qualitative analysis comparing individuals with TBI and a comparison group. METHODS AND PROCEDURES: Twenty-one individuals were interviewed using a semi-structured format focusing on various aspects of their partner relationship. A qualitative analysis of transcripts identified general themes. MAIN OUTCOMES AND RESULTS: Variability of relationship experiences (relationship relatedness and persistence of the partnership) was shown across groups. Aspects of relationship commitment to the partnership were revealed. Many individuals with TBI and their partners attributed various relationship changes to ageing and age-related issues and not to the TBI. CONCLUSIONS: These findings suggest that the effects of TBI are not clearly distinguished from age-related causes for change in the partners and their relationship. Older couples' experiences may be different from those of younger couples impacted by TBI, in the areas of sexuality, inter-personal support and role functions. PMID- 16243749 TI - Episodic pain syndrome restricted cheiro-oral region associated with pontine lesion. AB - Other than from the thalamus and sensory cortex, episodic pain is an extremely rare neurological manifestation in the central compartment. This study reports a middle-aged man who experienced an acute onset of episodic oscillatory burning pain restricted to the cheiro-oral region, who was found to have a singular infarct at the left tegmental pons. A close relationship between his pain attack and an elevation of arterial blood pressure was clearly observed. Blood adenohypophyseal hormones and electroencephalogram did not reveal an abnormality in the ictus. Neuroimaging and clinical studies did not support involvement of the thalamus, periaqueductal gray matter, hypothalamus or regional structure. Therefore, episodic pain may be an isolated manifestation with a pontine lesion. A relapsing expansion of focal cerebral oedema with fluid retention may have corresponded to the oscillation of his sensory deficit. This accumulating, devastating calamity by a repetitive and paroxysmal offense after a blood-brain barrier breakdown should be cautiously reviewed. PMID- 16243748 TI - The high-level mobility assessment tool (HiMAT) for traumatic brain injury. Part 1: Item generation. AB - PRIMARY OBJECTIVE: To generate a comprehensive list of items for a scale suitable for assessing high-level mobility in people with traumatic brain injury (TBI). RESEARCH DESIGN: High-level mobility items were generated following a critical evaluation of existing adult and paediatric mobility scales and by surveying expert clinicians for opinions about items appropriate for inclusion on a high level mobility scale. MAIN OUTCOMES AND RESULTS: The critical evaluation identified 18 different items from 31 mobility scales. These included nine walking items in addition to higher level activities such as stair use, running, jumping and hopping. Expert clinicians generated 157 items that were collated and condensed to 88 items for ranking on a questionnaire. Fifteen items on the questionnaire were rated as very important by 80% of the expert clinicians. These included walking forwards, walking on slopes and different surfaces, changing direction, walking long distances and stair use. Running items included forwards, backwards, on slopes and over different surfaces, changing direction, stopping and starting as well as running long distances. Balancing in single limb stance was also included. CONCLUSION: The final list comprised walking, running, hopping, skipping, jumping and balance items. This initial version of the HiMAT has face and content validity although requires further testing to investigate whether it is uni-dimensional and valid for people with TBI. PMID- 16243750 TI - Foreign accent syndrome following traumatic brain injury. AB - Foreign accent syndrome (FAS) is a very rare disorder, caused by lesions of the dominant brain hemisphere and defined as a loss of normal phonetic contrast when using the mother language. The pronunciation is perceived by native speakers as compromised by a foreign accent. This study reports about a 35-years old woman, with a FAS following a brain injury with a traumatic left temporal haemorrhage, who experienced excellent remission after 2 years. PMID- 16243751 TI - Homocysteine, folate, and congenital heart defects. AB - Congenital heart disease will be the next frontier for prevention by periconceptional management of homocysteine and its metabolites by folate supplementation. Evidence for the connection between maternal and fetal folate metabolism and congenital malformations of the cardiovascular system is reviewed including possible applications to the large population of patients at risk for a child with congenital heart disease. PMID- 16243752 TI - Levels of matrix metalloproteinase-9 and its inhibitor in guinea pig asthma model following ovalbumin challenge. AB - Our objective was to investigate the role of matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) and their cellular sources in guinea pig asthma model following ovalbumin (OA) challenge. Twenty guinea pigs were divided into 2 groups: OA-challenged group were sensitized by intraperitoneal injection with OA and exposed to OA while the control group used saline in a similar manner. The positive level index (PLI), defined as the percentage of positive-stained cells x average optical density, was used to assess the expressing levels of MMP-9 and TIMP-1. The levels of MMP-9 from OA challenged group were significantly higher than those of controls. However, no significant difference of the TIMP-1 levels between the OA-challenged group and controls were found. MMP-9 was expressed strongly in both inflammatory and structural cells while TIMP-1 was expressed weakly in inflammatory cells and structural cells. Our results implied that MMP-9 may contribute to tissue remodeling in asthma and both structural and inflammatory cells within lung tissue are the main cellular sources of MMP-9. Excessive expression of MMP-9 in the early or acute stage of asthma may be associated with the initiation of bronchial injury and inflammatory cells accumulation. PMID- 16243754 TI - Posttransplant de novo membranous nephropathy in childhood. AB - Posttransplant proteinuria is a recognized, but relatively uncommon, presentation of renal transplant dysfunction. Significant proteinuria occurs in around 10-15% of renal transplant recipients. We present a case of de novo posttransplant membranous nephropathy in childhood complicating renal transplantation for severe congenital obstructive uropathy and review the pathology, pathogenesis, and clinical implications of this condition. In the majority of cases, the cause of posttransplant proteinuria is either related to chronic allograft nephropathy or recurrence of the glomerulonephritis for which transplantation was indicated. In a minority, however, de novo posttransplant membranous nephropathy (DNPMN) is identified on biopsy. The histopathological findings in some cases may either be similar to those of classical membranous nephropathy, or may be more subtle, showing focal segmental variation in severity, often in conjuction with the features of chronic allograft nephropathy. The use of ancillary techniques including immunohistochemistry and electron microscopy may be required to confirm the diagnosis. The presence of posttransplant de novo membranous nephropathy may be associated with an increased risk of graft loss. PMID- 16243753 TI - Lethal infantile cortical hyperostosis. AB - We present an unusual case of the rare occurrence of lethal infantile cortical hyperostosis (Caffey disease). Clinical and Pathological aspects are discussed. PMID- 16243755 TI - Hypoxic-ischemic encephalopathy in infants: new challenges. AB - The subject of hypoxic-ischemic encephalopathy (HIE) is hardly new and yet remains an every day challenge to clinicians, mainly neonatologists and child neurologists. However, technological advances in neuroimaging do help in the diagnosis, but prevention of hypoxic-ischemic insults, ability to change the course of the disease, and treatment of the devastating consequences of HIE are lagging behind. With these issues in mind, we reviewed the literature on this subject and our own clinical experience in search of new developments in this area. The pathology and clinicopathologic correlates of HIE are emphasized as well. PMID- 16243756 TI - Segmentation defects of the human axial skeleton without dysostoses or skeletal dysplasias. AB - The incidence and associations of axial segmentation defects were examined in a series of perinatal autopsies. Variations in vertebral morphology (whether changes in overall numbers or homeotic transformations from one morphological type to another) and of number of well-formed ribs were common, and both were associated with extraskeletal malformations. Rudimentary cervical ribs were present in almost half of all cases and were not specifically associated with extraskeletal abnormalities. Radiographs of all fetuses and infants at autopsy are recommended as they may uncover axial skeletal abnormalities. PMID- 16243759 TI - Asynchronous colonization of Madagascar by the four endemic clades of primates, tenrecs, carnivores, and rodents as inferred from nuclear genes. AB - Madagascar harbors four large adaptive radiations of endemic terrestrial mammals: lemurs, tenrecs, carnivorans, and rodents. These rank among the most spectacular examples of evolutionary diversification, but their monophyly and origins are debated. The lack of Tertiary fossils from Madagascar leaves molecular studies as most promising to solve these controversies. We provide a simultaneous reconstruction of phylogeny and age of the four radiations based on a 3.5-kb data set from three nuclear genes (ADRA2B, vWF, and AR). The analysis supports each as a monophyletic clade, sister to African taxa, and thereby identifies four events of colonization out of Africa. To infer the time windows for colonization, we take into account both the divergence from the closest non-insular sister group and the initial intra-insular radiation, which is a novel but conservative approach in studies of the colonization history of Madagascar. We estimate that lemurs colonized Madagascar between 60 million years ago (Mya) (split from lorises) and 50 Mya (lemur radiation) (70-41 Mya taking 95% credibility intervals into account), tenrecs between 42 and 25 Mya (50-20 Mya), carnivorans between 26 and 19 Mya (33-14 Mya), and rodents between 24 and 20 Mya (30-15 Mya). These datings suggest at least two asynchronous colonization events: by lemurs in the Late Cretaceous-Middle Eocene, and by carnivorans and rodents in the Early Oligocene-Early Miocene. The colonization by tenrecs may have taken place simultaneously with either of these two events, or in a third event in the Late Eocene-Oligocene. Colonization by at least lemurs, rodents, and carnivorans appears to have occurred by overseas rafting rather than via a land bridge hypothesized to have existed between 45 and 26 Mya, but the second scenario cannot be ruled out if credibility intervals are taken into account. PMID- 16243760 TI - Hylid frog phylogeny and sampling strategies for speciose clades. AB - How should characters and taxa be sampled to resolve efficiently the phylogeny of ancient and highly speciose groups? We addressed this question empirically in the treefrog family Hylidae, which contains > 800 species and may be nonmonophyletic with respect to other anuran families. We sampled 81 species (54 hylids and 27 outgroups) for two mitochondrial genes (12S, ND1), two nuclear genes (POMC, c myc), and morphology (144 characters) in an attempt to resolve higher-level relationships. We then added 117 taxa to the combined data set, many of which were sampled for only one gene (12S). Despite the relative incompleteness of the majority of taxa, the resulting trees placed all taxa in the expected higher level clades with strong support, despite some taxa being > 90% incomplete. Furthermore, we found no relationship between the completeness of a taxon and the support (parsimony bootstrap or Bayesian posterior probabilities) for its localized placement on the tree. Separate analysis of the data set with the most taxa (12S) gives a somewhat problematic estimate of higher-level relationships, suggesting that data sets scored only for some taxa (ND1, nuclear genes, morphology) are important in determining the outcome of the combined analysis. The results show that hemiphractine hylids are not closely related to other hylids and should be recognized as a distinct family. They also show that the speciose genus Hyla is polyphyletic, but that its species can be arranged into three monophyletic genera. A new classification of hylid frogs is proposed. Several potentially misleading signals in the morphological data are discussed. PMID- 16243761 TI - Can incomplete taxa rescue phylogenetic analyses from long-branch attraction? AB - Taxon sampling may be critically important for phylogenetic accuracy because adding taxa can help to subdivide misleading long branches. Although the idea that added taxa can break up long branches was exemplified by a study of "incomplete" fossil taxa, the issue of taxon completeness (i.e., proportion of missing data) has been largely ignored in most subsequent discussions of taxon sampling and long-branch attraction. In this article, I use simulations to test the ability of incomplete taxa to subdivide long branches and improve phylogenetic accuracy in situations of potential long-branch attraction. The results show that for most methods and conditions examined, adding taxa that are only 50% complete may provide similar benefits to adding the same number of complete taxa (suggesting that the advantages of increased taxon sampling may be obtained with less data than previously considered). For parsimony, taxa that are less complete (5% to 25% complete) may often have limited ability to rescue analyses from long-branch attraction. In contrast, highly incomplete taxa can be surprisingly beneficial when using model-based methods. The results also suggest the importance of model-based methods in phylogenetic analyses that combine molecular and fossil data. PMID- 16243762 TI - An empirical assessment of long-branch attraction artefacts in deep eukaryotic phylogenomics. AB - In the context of exponential growing molecular databases, it becomes increasingly easy to assemble large multigene data sets for phylogenomic studies. The expected increase of resolution due to the reduction of the sampling (stochastic) error is becoming a reality. However, the impact of systematic biases will also become more apparent or even dominant. We have chosen to study the case of the long-branch attraction artefact (LBA) using real instead of simulated sequences. Two fast-evolving eukaryotic lineages, whose evolutionary positions are well established, microsporidia and the nucleomorph of cryptophytes, were chosen as model species. A large data set was assembled (44 species, 133 genes, and 24,294 amino acid positions) and the resulting rooted eukaryotic phylogeny (using a distant archaeal outgroup) is positively misled by an LBA artefact despite the use of a maximum likelihood-based tree reconstruction method with a complex model of sequence evolution. When the fastest evolving proteins from the fast lineages are progressively removed (up to 90%), the bootstrap support for the apparently artefactual basal placement decreases to virtually 0%, and conversely only the expected placement, among all the possible locations of the fast-evolving species, receives increasing support that eventually converges to 100%. The percentage of removal of the fastest evolving proteins constitutes a reliable estimate of the sensitivity of phylogenetic inference to LBA. This protocol confirms that both a rich species sampling (especially the presence of a species that is closely related to the fast evolving lineage) and a probabilistic method with a complex model are important to overcome the LBA artefact. Finally, we observed that phylogenetic inference methods perform strikingly better with simulated as opposed to real data, and suggest that testing the reliability of phylogenetic inference methods with simulated data leads to overconfidence in their performance. Although phylogenomic studies can be affected by systematic biases, the possibility of discarding a large amount of data containing most of the nonphylogenetic signal allows recovering a phylogeny that is less affected by systematic biases, while maintaining a high statistical support. PMID- 16243763 TI - Resolving deep phylogenetic relationships in salamanders: analyses of mitochondrial and nuclear genomic data. AB - Phylogenetic relationships among salamander families illustrate analytical challenges inherent to inferring phylogenies in which terminal branches are temporally very long relative to internal branches. We present new mitochondrial DNA sequences, approximately 2,100 base pairs from the genes encoding ND1, ND2, COI, and the intervening tRNA genes for 34 species representing all 10 salamander families, to examine these relationships. Parsimony analysis of these mtDNA sequences supports monophyly of all families except Proteidae, but yields a tree largely unresolved with respect to interfamilial relationships and the phylogenetic positions of the proteid genera Necturus and Proteus. In contrast, Bayesian and maximum-likelihood analyses of the mtDNA data produce a topology concordant with phylogenetic results from nuclear-encoded rRNA sequences, and they statistically reject monophyly of the internally fertilizing salamanders, suborder Salamandroidea. Phylogenetic simulations based on our mitochondrial DNA sequences reveal that Bayesian analyses outperform parsimony in reconstructing short branches located deep in the phylogenetic history of a taxon. However, phylogenetic conflicts between our results and a recent analysis of nuclear RAG-1 gene sequences suggest that statistical rejection of a monophyletic Salamandroidea by Bayesian analyses of our mitochondrial genomic data is probably erroneous. Bayesian and likelihood-based analyses may overestimate phylogenetic precision when estimating short branches located deep in a phylogeny from data showing substitutional saturation; an analysis of nucleotide substitutions indicates that these methods may be overly sensitive to a relatively small number of sites that show substitutions judged uncommon by the favored evolutionary model. PMID- 16243764 TI - An empirical examination of the utility of codon-substitution models in phylogeny reconstruction. AB - Models of codon substitution have been commonly used to compare protein-coding DNA sequences and are particularly effective in detecting signals of natural selection acting on the protein. Their utility in reconstructing molecular phylogenies and in dating species divergences has not been explored. Codon models naturally accommodate synonymous and nonsynonymous substitutions, which occur at very different rates and may be informative for recent and ancient divergences, respectively. Thus codon models may be expected to make an efficient use of phylogenetic information in protein-coding DNA sequences. Here we applied codon models to 106 protein-coding genes from eight yeast species to reconstruct phylogenies using the maximum likelihood method, in comparison with nucleotide- and amino acid-based analyses. The results appeared to confirm that expectation. Nucleotide-based analysis, under simplistic substitution models, were efficient in recovering recent divergences whereas amino acid-based analysis performed better at recovering deep divergences. Codon models appeared to combine the advantages of amino acid and nucleotide data and had good performance at recovering both recent and deep divergences. Estimation of relative species divergence times using amino acid and codon models suggested that translation of gene sequences into proteins led to information loss of from 30% for deep nodes to 66% for recent nodes. Although computational burden makes codon models unfeasible for tree search in large data sets, we suggest that they may be useful for comparing candidate trees. Nucleotide models that accommodate the differences in evolutionary dynamics at the three codon positions also performed well, at much less computational cost. We discuss the relationship between a model's fit to data and its utility in phylogeny reconstruction and caution against use of overly complex substitution models. PMID- 16243765 TI - An application of tissue and DNA banking for genomics and conservation: the Ambrose Monell Cryo-Collection (AMCC). PMID- 16243766 TI - Measuring support and finding unsupported relationships in supertrees. PMID- 16243767 TI - On probability and systematics: possibility, probability, and phylogenetic inference. PMID- 16243768 TI - DNA barcoding: perspectives from a "Partnerships for Enhancing Expertise in Taxonomy" (PEET) debate. PMID- 16243769 TI - The perils of DNA barcoding and the need for integrative taxonomy. PMID- 16243770 TI - The promise of DNA barcoding for taxonomy. PMID- 16243773 TI - Early epidermal destruction with subsequent epidermal hyperplasia is a unique feature of the papilloma-independent squamous cell carcinoma phenotype in PKCepsilon overexpressing transgenic mice. AB - Protein kinase C epsilon (PKCepsilon) overexpressing transgenic (PKCepsilon Tg) mice develop papilloma-independent squamous cell carcinomas (SCC) elicited by 7,12-dimethylbenz[a]anthracene (DMBA) tumor initiation and 12-O tetradecanoylphorbol-13-acetate (TPA) tumor promotion. We examined whether epidermal cell turnover kinetics was altered during the development of SCC in PKCepsilon Tg mice. Dorsal skin samples were fixed for histological examination. A single application of TPA resulted in extensive infiltration of polymorphonuclear neutrophils (PMNs) into the epidermis at 24 h after TPA treatment in PKCepsilon Tg mice while wild-type (WT) mouse skin showed focal infiltration by PMNs. Complete epidermal necrosis was observed at 48 h in PKCepsilon Tg mice only; at 72 h, epidermal cell regeneration beginning from hair follicles was observed in PKCepsilon Tg mice. Since the first TPA treatment to DMBA-initiated PKCepsilon Tg mouse skin led to epidermal destruction analogous to skin abrasion, we propose the papilloma-independent phenotype may be explained by death of initiated interfollicular cells originally destined to become papillomas. Epidermal destruction did not occur after multiple doses of TPA, presumably reflecting adaptation of epidermis to chronic TPA treatment. Prolonged hyperplasia in the hair follicle may result in the early neoplastic lesions originally described by Jansen et al. (2001) by expanding initiated cells in the hair follicles resulting in the subsequent development of SCC. PMID- 16243774 TI - Transgenic disruption of gap junctional intercellular communication enhances early but not late stage hepatocarcinogenesis in the rat. AB - Much experimental evidence supports the conclusion that loss of gap junctional intercellular communication (GJIC) contributes to carcinogenesis. Transgenic rats featuring a dominant negative mutant of the connexin 32 gene under albumin promoter control (Cx32Delta Tg-High and Cx32Delta Tg-Low lines, respectively with high and low copy numbers of the transgene) have disrupted GJIC, as demonstrated by scrape dye-transfer assay in vivo as previous report by Asamoto et al. (2004). In the present study, we investigated the susceptibility of these transgenic rats to a single intraperitoneal administration of diethylnitrosamine (DEN), and found a significant increase in preneoplastic glutathione S-transferase placental form (GST-P) positive lesions in the livers of Cx32Delta Tg-High but not Cx32Delta Tg Low rats. However, incidences of adenomas and hepatocellular carcinomas were not elevated at the end of the experiment (52 weeks). In addition, we investigated the promotional effect of phenobarbital (PB) on Cx32Delta Tg-High rats pretreated with DEN and found enhanced formation of GST-P positive lesions, in contrast to the lack of promoting effects reported for Cx32 deficient mice. The results indicate that although both high and low expression of the dominant negative connexin 32 mutant gene in our rats is able to inhibit gap junctional capacity, only high expression is effective at enhancing susceptibility to early stage DEN induced liver carcinogenesis. PMID- 16243775 TI - Safety of recombinant human factor XIII in a cynomolgus monkey model of extracorporeal blood circulation. AB - Factor XIII (FXIII) is a thrombin-activated plasma coagulation factor critical for blood clot stabilization and longevity. Administration of exogenous FXIII to replenish depleted stores after major surgery, including cardiopulmonary bypass, may reduce bleeding complications and transfusion requirements. Thus, a model of extracorporeal circulation (ECC) was developed in adult male cynomolgus monkeys (Macaca fascicularis) to evaluate the nonclinical safety of recombinant human FXIII (rFXIII). The hematological and coagulation profile in study animals during and after 2 h of ECC was similar to that reported for humans during and after cardiopulmonary bypass, including observations of anemia, thrombocytopenia, and activation of coagulation and platelets. Intravenous slow bolus injection of 300 U/kg (2.1 mg/kg) or 1000 U/kg (7 mg/kg) rFXIII after 2 h of ECC was well tolerated in study animals, and was associated with a dose-dependent increase in FXIII activity. No clinically significant effects in respiration, ECG, heart rate, blood pressure, body temperature, clinical chemistry, hematology (including platelet counts), or indicators of thrombosis (thrombin:anti-thrombin complex and D-Dimer) or platelet activation (platelet factor 4 and beta-thromboglobulin) were related to rFXIII administration. Specific examination of brain, heart, lung, liver, and kidney from rFXIII-treated animals provided no evidence of histopathological alterations suggestive of subclinical hemorrhage or thrombosis. Taken as a whole, the results demonstrate the ECC model suitably replicated the clinical presentation reported for humans during and after cardiopulmonary bypass surgery, and do not suggest significant concerns regarding use of rFXIII in replacement therapy after extracorporeal circulation. PMID- 16243776 TI - New experimental approaches for investigating interactions between Pyrococcus furiosus carbamate kinase and carbamoyltransferases, enzymes involved in the channeling of thermolabile carbamoyl phosphate. AB - A somewhat neglected but essential aspect of the molecular physiology of hyperthermophiles is the protection of thermolabile metabolites and coenzymes. An example is carbamoyl phosphate (CP), a precursor of pyrimidines and arginine, which is an extremely labile and potentially toxic intermediate. The first evidence for a biologically significant interaction between carbamate kinase (CK) and ornithine carbamoyltransferase (OTC) from Pyrococcus furiosus was provided by affinity electrophoresis and co-immunoprecipitation in combination with cross linking (Massant et al. 2002). Using the yeast two-hybrid system, Hummel-Dreyer chromatography and isothermal titration calorimetry, we obtained additional concrete evidence for an interaction between CK and OTC, the first evidence for an interaction between CK and aspartate carbamoyltransferase (ATC) and an estimate of the binding constant between CK and ATC. The physical interaction between CK and OTC or ATC may prevent thermodenaturation of CP in the aqueous cytoplasmic environment. Here we emphasize the importance of developing experimental approaches to investigate the mechanism of thermal protection of metabolic intermediates by metabolic channeling and the molecular basis of transient protein-protein interactions in the physiology of hyperthermophiles. PMID- 16243777 TI - Characterization of the Family I inorganic pyrophosphatase from Pyrococcus horikoshii OT3. AB - A gene encoding for a putative Family I inorganic pyrophosphatase (PPase, EC 3.6.1.1) from the hyperthermophilic archaeon Pyrococcus horikoshii OT3 was cloned and the biochemical characteristics of the resulting recombinant protein were examined. The gene (Accession No. 1907) from P. horikoshii showed some identity with other Family I inorganic pyrophosphatases from archaea. The recombinant PPase from P. horikoshii (PhPPase) has a molecular mass of 24.5 kDa, determined by SDS-PAGE. This enzyme specifically catalyzed the hydrolysis of pyrophosphate and was sensitive to NaF. The optimum temperature and pH for PPase activity were 70 degrees C and 7.5, respectively. The half-life of heat inactivation was about 50 min at 105 degrees C. The heat stability of PhPPase was enhanced in the presence of Mg2+. A divalent cation was absolutely required for enzyme activity, Mg2+ being most effective; Zn2+, Co2+ and Mn2+ efficiently supported hydrolytic activity in a narrow range of concentrations (0.05-0.5 mM). The K(m) for pyrophosphate and Mg2+ were 113 and 303 microM, respectively; and maximum velocity, V(max), was estimated at 930 U mg(-1). PMID- 16243778 TI - Role of the precorrin 6-X reductase gene in cobamide biosynthesis in Methanococcus maripaludis. AB - In Methanococcus maripaludis strain JJ, deletion of the homolog to cbiJ, which encodes the corrin biosynthetic enzyme precorrin 6-X reductase, yielded an auxotroph that required either cobamide or acetate for good growth. This phenotype closely resembled that of JJ117, a mutant in which tandem repeats were introduced into the region immediately downstream of the homolog of cbiJ. Mutant JJ117 also produced low quantities of cobamides, about 15 nmol g(-1) protein or 1 2% of the amount found in wild-type cells. These results confirm the role of the cbiJ homolog in cobamide biosynthesis in the Archaea and suggest the presence of low amounts of a bypass activity in these organisms. PMID- 16243779 TI - Succession of methanogenic archaea in rice straw incorporated into a Japanese rice field: estimation by PCR-DGGE and sequence analyses. AB - The succession and phylogenetic profiles of methanogenic archaeal communities associated with rice straw decomposition in rice-field soil were studied by polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) analysis followed by 16S rDNA sequencing. Nylon bags containing either leaf sheaths or blades were buried in the plowed layer of a Japanese rice field under drained conditions during the off-crop season and under flooded conditions after transplanting. In addition, rice straw samples that had been buried in the rice field under drained conditions during the off-crop season were temporarily removed during spring plowing and then re-buried in the same rice field under flooded conditions at transplanting. Populations of methanogenic archaea were examined by amplification of the 16S rRNA genes in the DNA extracted from the rice straw samples. No PCR product was produced for samples of leaf sheath or blade prior to burial or after burial under drained conditions, indicating that the methanogen population was very small during decomposition of rice straw under oxic conditions. Many common bands were observed in rice straw samples of leaf sheath and blade during decomposition of rice straw under flooded conditions. Cluster analysis based on DGGE patterns divided methanogenic archaeal communities into two groups before and after the mid-season drainage. Sequence analysis of DGGE bands that were commonly present were closely related to Methanomicrobiales and Rice cluster I. Methanomicrobiales, Rice cluster I and Methanosarcinales were major members before the mid-season drainage, whereas the DGGE bands that characterized methanogenic archaeal communities after the mid-season drainage were closely related to Methanomicrobiales. These results indicate that mid season drainage affected the methanogenic archaeal communities irrespective of their location on rice straw (sheath and blade) and the previous history of decomposition during the off-crop season. PMID- 16243780 TI - A study of archaeal enzymes involved in polar lipid synthesis linking amino acid sequence information, genomic contexts and lipid composition. AB - Cellular membrane lipids, of which phospholipids are the major constituents, form one of the characteristic features that distinguish Archaea from other organisms. In this study, we focused on the steps in archaeal phospholipid synthetic pathways that generate polar lipids such as archaetidylserine, archaetidylglycerol, and archaetidylinositol. Only archaetidylserine synthase (ASS), from Methanothermobacter thermautotrophicus, has been experimentally identified. Other enzymes have not been fully examined. Through database searching, we detected many archaeal hypothetical proteins that show sequence similarity to members of the CDP alcohol phosphatidyltransferase family, such as phosphatidylserine synthase (PSS), phosphatidylglycerol synthase (PGS) and phosphatidylinositol synthase (PIS) derived from Bacteria and Eukarya. The archaeal hypothetical proteins were classified into two groups, based on the sequence similarity. Members of the first group, including ASS from M. thermautotrophicus, were closely related to PSS. The rough agreement between PSS homologue distribution within Archaea and the experimentally identified distribution of archaetidylserine suggested that the hypothetical proteins are ASSs. We found that an open reading frame (ORF) tends to be adjacent to that of ASS in the genome, and that the order of the two ORFs is conserved. The sequence similarity of phosphatidylserine decarboxylase to the product of the ORF next to the ASS gene, together with the genomic context conservation, suggests that the ORF encodes archaetidylserine decarboxylase, which may transform archaetidylserine to archaetidylethanolamine. The second group of archaeal hypothetical proteins was related to PGS and PIS. The members of this group were subjected to molecular phylogenetic analysis, together with PGSs and PISs and it was found that they formed two distinct clusters in the molecular phylogenetic tree. The distribution of members of each cluster within Archaea roughly corresponded to the experimentally identified distribution of archaetidylglycerol or archaetidylinositol. The molecular phylogenetic tree patterns and the correspondence to the membrane compositions suggest that the two clusters in this group correspond to archaetidylglycerol synthases and archaetidylinositol synthases. No archaeal hypothetical protein with sequence similarity to known phosphatidylcholine synthases was detected in this study. PMID- 16243781 TI - Oligomerization of Sulfolobus solfataricus signature amidase is promoted by acidic pH and high temperature. AB - The recombinant amidase from the hyperthermophylic archaeon Sulfolobus solfataricus (SSAM) a signature amidase, was cloned, purified and characterized. The enzyme is active on a large number of aliphatic and aromatic amides over the temperature range 60-95 degrees C and at pH values between 4.0 and 9.5, with an optimum at pH 5.0. The recombinant enzyme is in the form of a dimer of about 110 kD that reversibly associates into an octamer in a pH-dependent reaction. The pH dependence of the state of association was studied using gel permeation chromatography, analytical ultracentrifugation and dynamic light scattering techniques. At pH 7.0 all three techniques show the presence of two species, in about equal amounts, which is compatible with the existence of a dimeric and an octameric form. In decreasing pH, the dimers formed the octameric species and in increasing pH, the octameric species was converted to dimers. Above pH 8.0, only dimers were present, below pH 3.0 only octamers were present. The association of dimers into octamers decreased in non-polar solvents and increased with temperature. A mutant (Y41C) was obtained that did not show this behavior. PMID- 16243782 TI - Application of single molecule technology to rapidly map long DNA and study the conformation of stretched DNA. AB - Herein we describe the first application of direct linear analysis (DLA) to the mapping of a bacterial artificial chromosome (BAC), specifically the 185.1 kb long BAC 12M9. DLA is a single molecule mapping technology, based on microfluidic elongation and interrogation of individual DNA molecules, sequence-specifically tagged with bisPNAs. A DNA map with S/N ratio sufficiently high to detect all major binding sites was obtained using only 200 molecule traces. A new method was developed to extract an oriented map from an averaged map that included a mixture of head-first and tail-first DNA traces. In addition, we applied DLA to study the conformation and tagging of highly stretched DNA. Optimal conditions for promoting sequence-specific binding of bisPNA to an 8 bp target site were elucidated using DLA, which proved superior to electromobility shift assays. DLA was highly reproducible with a hybridized tag position localized with an accuracy of +/-0.7 microm or +/-2.1 kb demonstrating its utility for rapid mapping of large DNA at the single molecule level. Within this accuracy, DNA molecules, stretched to at least 85% of their contour length, were stretched uniformly, so that the map expressed in relative coordinates, was the same regardless of the molecule extension. PMID- 16243783 TI - Draft versus finished sequence data for DNA and protein diagnostic signature development. AB - Sequencing pathogen genomes is costly, demanding careful allocation of limited sequencing resources. We built a computational Sequencing Analysis Pipeline (SAP) to guide decisions regarding the amount of genomic sequencing necessary to develop high-quality diagnostic DNA and protein signatures. SAP uses simulations to estimate the number of target genomes and close phylogenetic relatives (near neighbors or NNs) to sequence. We use SAP to assess whether draft data are sufficient or finished sequencing is required using Marburg and variola virus sequences. Simulations indicate that intermediate to high-quality draft with error rates of 10(-3)-10(-5) (approximately 8x coverage) of target organisms is suitable for DNA signature prediction. Low-quality draft with error rates of approximately 1% (3x to 6x coverage) of target isolates is inadequate for DNA signature prediction, although low-quality draft of NNs is sufficient, as long as the target genomes are of high quality. For protein signature prediction, sequencing errors in target genomes substantially reduce the detection of amino acid sequence conservation, even if the draft is of high quality. In summary, high-quality draft of target and low-quality draft of NNs appears to be a cost effective investment for DNA signature prediction, but may lead to underestimation of predicted protein signatures. PMID- 16243784 TI - Structural insights into abasic site for Fpg specific binding and catalysis: comparative high-resolution crystallographic studies of Fpg bound to various models of abasic site analogues-containing DNA. AB - Fpg is a DNA glycosylase that recognizes and excises the mutagenic 8-oxoguanine (8-oxoG) and the potentially lethal formamidopyrimidic residues (Fapy). Fpg is also associated with an AP lyase activity which successively cleaves the abasic (AP) site at the 3' and 5' sides by betadelta-elimination. Here, we present the high-resolution crystal structures of the wild-type and the P1G defective mutant of Fpg from Lactococcus lactis bound to 14mer DNA duplexes containing either a tetrahydrofuran (THF) or 1,3-propanediol (Pr) AP site analogues. Structures show that THF is less extrahelical than Pr and its backbone C5'-C4'-C3' diverges significantly from those of Pr, rAP, 8-oxodG and FapydG. Clearly, the heterocyclic oxygen of THF is pushed back by the carboxylate of the strictly conserved E2 residue. We can propose that the ring-opened form of the damaged deoxyribose is the structure active form of the sugar for Fpg catalysis process. Both structural and functional data suggest that the first step of catalysis mediated by Fpg involves the expulsion of the O4' leaving group facilitated by general acid catalysis (involving E2), rather than the immediate cleavage of the N-glycosic bond of the damaged nucleoside. PMID- 16243785 TI - Specific function of a plastid sigma factor for ndhF gene transcription. AB - The complexity of the plastid transcriptional apparatus (two or three different RNA polymerases and numerous regulatory proteins) makes it very difficult to attribute specific function(s) to its individual components. We have characterized an Arabidopsis T-DNA insertion line disrupting the nuclear gene coding for one of the six plastid sigma factors (SIG4) that regulate the activity of the plastid-encoded RNA polymerase PEP. This mutant shows a specific diminution of transcription of the plastid ndhF gene, coding for a subunit of the plastid NDH [NAD(P)H dehydrogenase] complex. The absence of another NDH subunit, i.e. NDHH, and the absence of a chlorophyll fluorescence transient previously attributed to the activity of the plastid NDH complex indicate a strong down regulation of NDH activity in the mutant plants. Results suggest that plastid NDH activity is regulated on the transcriptional level by an ndhF-specific plastid sigma factor, SIG4. PMID- 16243786 TI - Use of altered-specificity binding Oct-4 suggests an absence of pluripotent cell specific cofactor usage. AB - Oct-4 is a POU domain transcription factor that is critical for maintaining pluripotency and for stem cell renewal. Previous studies suggest that transcription regulation by Oct-4 at particular enhancers requires the input of a postulated E1A-like cofactor that is specific to pluripotent cells. However, such studies have been limited to the use of enhancer elements that bind other POU protein family members in addition to Oct-4, thus preventing a 'clean' assessment of any Oct-4:cofactor relationships. Other attempts to study Oct-4 functionality in a more 'stand-alone' situation target Oct-4 transactivation domains to DNA using heterologous binding domains, a methodology which is known to generate artificial data. To circumvent these issues, an altered-specificity binding Oct-4 (Oct-4RR) and accompanying binding site, which binds Oct-4RR only, were generated. This strategy has previously been shown to maintain Oct-1:cofactor interactions that are highly binding-site and protein/binding conformation specific. This system therefore allows a stand-alone study of Oct-4 function in pluripotent versus differentiated cells, without interference from endogenous POU factors and with minimal deviation from bound wild-type protein characteristics. Subsequently, it was demonstrated that Oct-4RR and the highly transactive regions of its N-terminus determined here, and its C-terminus, have the same transactivation profile in pluripotent and differentiated cells, thus providing strong evidence against the existence of such a pluripotent cell-specific Oct-4 cofactor. PMID- 16243787 TI - Function of the ribosomal E-site: a mutagenesis study. AB - Ribosomes synthesize proteins according to the information encoded in mRNA. During this process, both the incoming amino acid and the nascent peptide are bound to tRNA molecules. Three binding sites for tRNA in the ribosome are known: the A-site for aminoacyl-tRNA, the P-site for peptidyl-tRNA and the E-site for the deacylated tRNA leaving the ribosome. Here, we present a study of Escherichia coli ribosomes with the E-site binding destabilized by mutation C2394G of the 23S rRNA. Expression of the mutant 23S rRNA in vivo caused increased frameshifting and stop codon readthrough. The progression of these ribosomes through the ribosomal elongation cycle in vitro reveals ejection of deacylated tRNA during the translocation step or shortly after. E-site compromised ribosomes can undergo translocation, although in some cases it is less efficient and results in a frameshift. The mutation affects formation of the P/E hybrid site and leads to a loss of stimulation of the multiple turnover GTPase activity of EF-G by deacylated tRNA bound to the ribosome. PMID- 16243788 TI - Epidermal growth factor receptor inhibitors: a moving target? PMID- 16243789 TI - Multifocal renal cancer: genetic basis and its medical relevance. PMID- 16243790 TI - Gene expression profiles of B-lineage adult acute lymphocytic leukemia reveal genetic patterns that identify lineage derivation and distinct mechanisms of transformation. AB - PURPOSE: To characterize gene expression signatures in acute lymphocytic leukemia (ALL) cells associated with known genotypic abnormalities in adult patients. EXPERIMENTAL DESIGN: Gene expression profiles from 128 adult patients with newly diagnosed ALL were characterized using high-density oligonucleotide microarrays. All patients were enrolled in the Italian GIMEMA multicenter clinical trial 0496 and samples had >90% leukemic cells. Uniform phenotypic, cytogenetic, and molecular data were also available for all cases. RESULTS: T-lineage ALL was characterized by a homogeneous gene expression pattern, whereas several subgroups of B-lineage ALL were evident. Within B-lineage ALL, distinct signatures were associated with ALL1/AF4 and E2A/PBX1 gene rearrangements. Expression profiles associated with ALL1/AF4 and E2A/PBX1 are similar in adults and children. BCR/ABL+ gene expression pattern was more heterogeneous and was most similar to ALL without known molecular rearrangements. We also identified a set of 83 genes that were highly expressed in leukemia blasts from patients without known molecular abnormalities who subsequently relapsed following therapy. Supervised analysis of kinase genes revealed a high-level FLT3 expression in a subset of cases without molecular rearrangements. Two other kinases (PRKCB1 and DDR1) were highly expressed in cases without molecular rearrangements, as well as in BCR/ABL positive ALL. CONCLUSIONS: Genomic signatures are associated with phenotypically and molecularly well defined subgroups of adult ALL. Genomic profiling also identifies genes associated with poor outcome in cases without molecular aberrations and specific genes that may be new therapeutic targets in adult ALL. PMID- 16243791 TI - Carbonic anhydrase IX expression and tumor oxygenation status do not correlate at the microregional level in locally advanced cancers of the uterine cervix. AB - PURPOSE: Carbonic anhydrase IX (CA IX) can be induced by hypoxia in vitro and shows an immunohistochemical expression pattern that is predominantly found in perinecrotic tumor areas and correlates with exogenous markers of hypoxia, such as pimonidazole. CA IX might therefore serve as an endogenous marker of tumor hypoxia, although comparisons of CA IX immunostaining with direct oxygenation measurements using pO2 microsensors have thus far yielded contradictory results. EXPERIMENTAL DESIGN: Because tumor heterogeneity may be among the factors responsible for the discrepancy between the two methods, CA IX expression in tissue samples originating from oxygen microelectrode tracks of locally advanced cervical cancers was assessed in this study. Seventy-seven biopsy specimens were analyzed immunohistochemically using an anti-CA IX rabbit polyclonal antibody and semiquantitative scoring. RESULTS: CA IX expression showed no correlation with the oxygenation variables median pO2 and hypoxic fraction 2.5, 5, or 10. Cases with higher International Federation of Gynecology and Obstetrics stages (IIb IVa) exhibited stronger expression of CA IX (P = 0.035) and CA IX expression tended to be more prevalent in node-positive patients (P = 0.051). CONCLUSIONS: These data indicate that CA IX cannot be recommended as a substitute for oxygen microelectrode measurements. That the expression of CA IX does not correlate with the oxygenation status may be due to the degree to which other factors, such as nutrient (e.g., glucose) deficiency or the action of oncogenic mutations, can modulate the in vivo expression of this protein, rendering a strict association with tumor hypoxia too unreliable for clinical use. PMID- 16243792 TI - Molecular genetic evidence for the independent origin of multifocal papillary tumors in patients with papillary renal cell carcinomas. AB - PURPOSE: In patients with papillary renal cell carcinoma, it is not uncommon to find two or more anatomically distinct and histologically similar tumors at radical nephrectomy. Whether these multiple papillary lesions result from intrarenal metastasis or arise independently is unknown. Previous studies have shown that multifocal clear cell renal cell carcinomas express identical allelic loss and shift patterns in the different tumors within the same kidney, consistent with a clonal origin. However, similar clonality assays for multifocal papillary renal cell neoplasia have not been done. Molecular analysis of microsatellite and chromosome alterations and X-chromosome inactivation status in separate tumors in the same patient can be used to study the genetic relationships among the coexisting multiple tumors. EXPERIMENTAL DESIGN: We examined specimens from 21 patients who underwent radical nephrectomy for renal cell carcinoma. All patients had multiple separate papillary lesions (ranging from 2 to 5). Eighteen patients had multiple papillary renal cell carcinomas. Seven had one or more papillary renal cell carcinomas with coexisting papillary adenomas. Genomic DNA samples were prepared from formalin-fixed, paraffin embedded tissue sections using laser-capture microdissection. Loss of heterozygosity assays were done for six microsatellite polymorphic markers for putative tumor suppressor genes on chromosomes 3p14 (D3S1285), 7q31 (D7S522), 9p21 (D9S171), 16q23 (D16S507), 17q21 (D17S1795), and 17p13 (TP53). X-chromosome inactivation analyses were done on the papillary kidney tumors from three female patients. Fluorescence in situ hybridization analysis was done on the tumors of selected patients showing allelic loss at loci on chromosome 7 and/or chromosome 17. RESULTS: Twenty of 21 (95%) cases showed allelic loss in one or more of the papillary lesions in at least one of the six polymorphic markers analyzed. A concordant allelic loss pattern between each coexisting kidney tumor was seen in only 1 of 21 (5%) cases. A concordant pattern of nonrandom X-chromosome inactivation in the coexisting multiple papillary lesions was seen in two of three female patients. A discordant pattern of X-chromosome inactivation was seen in the tumors of the other female patient. Fluorescence in situ hybridization showed that the majority of tumors analyzed had gains of chromosomes 7 and 17. Two patients had one tumor with chromosomal gain and another separate tumor that did not. CONCLUSION: Our data suggest that, unlike multifocal clear cell renal cell carcinomas, the multiple tumors in patients with papillary renal cell carcinoma arise independently. Thus, intrarenal metastasis does not seem to play an important role in the spread of papillary renal cell carcinoma, a finding that has surgical, therapeutic, and prognostic implications. PMID- 16243793 TI - Novel genes associated with malignant melanoma but not benign melanocytic lesions. AB - PURPOSE: Cutaneous melanoma is a common, aggressive cancer with increasing incidence. The identification of melanoma-specific deregulated genes could provide molecular markers for lymph node staging assays and further insight into melanoma tumorigenesis. EXPERIMENTAL DESIGN: Total RNA isolated from 45 primary melanoma, 18 benign skin nevi, and 7 normal skin tissue specimens were analyzed on an Affymetrix Hu133A microarray containing 22,000 probe sets. RESULTS: Hierarchical clustering revealed a distinct separation of the melanoma samples from the benign and normal specimens. Novel genes associated with malignant melanoma were identified. Differential gene expression of two melanoma-specific genes, PLAB and L1CAM, were tested by a one-step quantitative reverse transcription-PCR assay on primary malignant melanoma, benign nevi, and normal skin samples, as well as on malignant melanoma lymph node metastasis and melanoma free lymph nodes. The performance of the markers was compared with conventional melanoma markers such as tyrosinase, gp100, and MART1. CONCLUSION: Our study systematically identified novel melanoma-specific genes and showed the feasibility of using a combination of PLAB and L1CAM in a reverse transcription PCR assay to differentiate clinically relevant samples containing benign or malignant melanocytes. PMID- 16243794 TI - Ovarian carcinomas: CCN genes are aberrantly expressed and CCN1 promotes proliferation of these cells. AB - PURPOSE: The connective tissue growth factor/cysteine-rich 61/nephroblastoma overexpressed (CCN) family consists of six matricellular proteins that are involved in various cellular functions, such as proliferation, development, and angiogenesis. The purpose of this study was to explore the possibility that CCN genes are involved in ovarian cancers. EXPERIMENTAL DESIGN: We quantified CCN expression in a series of 59 ovarian cancers using quantitative real-time reverse transcription-PCR. CCN1 protein levels were further determined by immunohistochemistry and Western blot analysis. Overexpression and inhibition of CCN1 expression by small interfering RNA were used to examine its role in ovarian cancer cell proliferation in vitro and in vivo. RESULTS: We found dysregulation of levels of the various CCN mRNAs in ovarian cancers compared with their expression in normal whole ovaries. Expression of CCN1 protein was detected in normal ovarian epithelial cells and ovarian tumors as well as in ovarian cancer cell lines. Furthermore, estrogen increased CCN1 mRNA and protein levels in ovarian cancer cells. Ectopic expression of CCN1 enhanced the growth of ovarian cancer cells in liquid culture, whereas inhibition of its expression decreased proliferation and increased apoptosis in these cells. The observed changes in cell growth were accompanied with activation of Akt and extracellular signal regulated kinase (ERK) signaling pathways. Stable expression of CCN1 in SKOV3 cells significantly increased tumorigenicity in nude mice. Finally, overexpression of CCN1 conferred resistant to carboplatin-induced apoptosis in SKOV3 cells. CONCLUSIONS: This is the first study to show abnormalities in CCN expression in ovarian carcinomas. Furthermore, our results suggest that CCN1 may play a role in ovarian carcinogenesis by stimulating survival and antiapoptotic signaling pathways. PMID- 16243795 TI - Expression of phosphorylated Ser70 of Bcl-2 correlates with malignancy in human colorectal neoplasms. AB - PURPOSE: Bcl-2 is a model apoptosis suppressor postulated to promote tumorigenesis. Recently, it has been reported that Bcl-2 undergoes phosphoregulation of its Ser70 to substantially alter its molecular function. Previous studies further suggest that such phospho-Bcl-2 regulation may influence tumor progression in colorectal and other cancers; however, phosphorylation status of the Ser70 of Bcl-2 (pSer70) in vivo in tumors remains obscure. To elucidate this question that may suggest the biological role, we molecularly screened a panel of human colorectal adenomas and adenocarcinomas for endogenous expression of pSer70 Bcl-2. EXPERIMENTAL DESIGN: An antibody specific against pSer70 Bcl-2 was generated for thorough immunohistochemical examination of paraffin-embedded tumor specimens, allowing detection of the endogenously expressed antigen among a range of Bcl-2-positive colorectal neoplasms, including 75 tubular adenomas, 114 adenocarcinomas, and 15 cases of cancer in adenomas. RESULTS: Loss of pSer70 Bcl-2 expression was observed in adenocarcinomas in a differentiation-dependent manner (positivities: well differentiated 63%, moderately differentiated 52%, and poorly differentiated 12%), whereas tubular adenomas maintained their expression (positivity 88%). Interestingly, an inverse correlation was found between expression of pSer70 Bcl-2 and Ki-67 antigen in those cases of cancer in adenoma (P < 0.01). It was further observed that loss of pSer70 Bcl-2 expression was associated with significantly shorter survival (P < 0.05) and correlated with clinical stages and lymph node metastasis (P < 0.05 and P < 0.05, respectively). CONCLUSIONS: Loss of pSer70 Bcl-2 expression is closely linked to biological aggressiveness in colorectal tumors and represents a statistically significant molecular index for prognosis of patients with these tumors. PMID- 16243796 TI - Macrophage migration inhibitory factor is a determinant of hypoxia-induced apoptosis in colon cancer cell lines. AB - Hypoxia contributes to cytotoxic chemotherapy and radiation resistance and may play a role in the efficacy of antiangiogenesis cancer therapy. We have generated a series of cell lines derived from the colon adenocarcinoma models HT29 and HCT116 by exposing cells in vitro to repeated sublethal periods of profound hypoxia. These cell lines have altered sensitivity to hypoxia-induced apoptosis: those derived from HT29 are resistant, whereas those from HCT116 are more susceptible. We used cDNA selected subtractive hybridization to identify novel genes mediating sensitivity to hypoxia-induced apoptosis and isolated macrophage migration inhibitory factor (MIF) from the hypoxia-conditioned cell lines. MIF expression correlates with susceptibility of the cell lines to apoptosis. In hypoxia-resistant cells, the induction of apoptosis by hypoxia can be restored by the addition of exogenous recombinant MIF protein, suggesting that resistance may result in part from down-regulation of MIF production possibly through an autocrine loop. Inhibition of MIF using small interfering RNA in the susceptible lines conferred resistance to hypoxia-induced cell death. The relative expression of MIF in the hypoxia-conditioned cells implanted s.c. in severe combined immunodeficient mice in vivo was similar to that observed in vitro. In an analysis of 12 unrelated colon tumor cell lines, MIF expression and response to hypoxia varied widely. Cell lines in which MIF was inducible by hypoxia were more sensitive to oxaliplatin. In human colon tumor specimens analyzed by immunohistochemistry, MIF expression was similarly variable. There was no detectable expression of MIF in normal colon mucosa or adenoma but positive staining in all carcinomas tested. Taken together, these data indicate that MIF may be a determinant of hypoxia-induced apoptosis in vitro and that its variable expression in human colon cancers may indicate a functional role in vivo. We suggest that MIF expression in colorectal cancer may be a marker of susceptibility to therapies that may depend on induction of hypoxia, possibly including antiangiogenic therapy. PMID- 16243797 TI - Molecular pathogenesis of ovarian borderline tumors: new insights and old challenges. AB - Ovarian borderline (low malignant potential) tumors are a puzzling group of neoplasms that do not fall neatly into benign or malignant categories. Their behavior is enigmatic, their pathogenesis unclear, and their clinical management controversial, especially for serous borderline tumors (SBT), the most common type of ovarian borderline tumor. Clarifying the nature of borderline tumors and their relationship to invasive carcinoma has puzzled investigators since the category was created over 30 years ago. Much of the confusion and controversy concerning these tumors is due to a lack of understanding of their pathogenesis and an absence of a model for the development of ovarian carcinoma. This review summarizes recent molecular studies of ovarian borderline tumors with special emphasis on the role of SBT in tumor progression and its relationship to ovarian serous carcinoma. PMID- 16243799 TI - In vitro and in vivo targeting properties of iodine-123- or iodine-131-labeled monoclonal antibody 14C5 in a non-small cell lung cancer and colon carcinoma model. AB - PURPOSE: The monoclonal antibody (mAb) 14C5 is a murine IgG1 directed against a yet undefined molecule involved in cell substrate adhesion found on the surface of malignant breast cancer tissue. mAb 14C5 is able to inhibit cell substrate adhesion and invasion of breast cancer cells in vitro. In normal tissues as well as in the stroma surrounding in situ carcinomas of the breast, no expression of the antigen 14C5 occurs. The aim of this study was to investigate the in vitro and in vivo targeting properties of 123I- and 131I-labeled mAb 14C5 as a novel agent for radioimmunodetection and radioimmunotherapy. EXPERIMENTAL DESIGN: Internalization of mAb 14C5 was investigated with 125I-labeled mAb 14C5 and by confocal laser scanning microscopy. Biodistribution studies of 131I-labeled mAb 14C5 and planar gamma imaging were done in nude mice bearing an A549 (non-small cell lung carcinoma) or a LoVo (colon carcinoma) tumor. RESULTS: Internalization studies with both A549 and LoVo cells showed that 125I-labeled mAb 14C5 is slowly internalized with approximately 30% of the initially bound mAb 14C5 internalized after 2 hours at 37 degrees C. Internalization of mAb 14C5 could be visualized with confocal laser scanning microscopy. In vivo, radioisotope uptake peaked at 24 hours for both tumor models (n = 5) with no significant difference in percentage of injected dose/g tissue (A549 10.4 +/- 0.8 and LoVo 9.3 +/- 0.8). Via planar gamma camera imaging, A549 lung tumors as well as LoVo colon tumors could be clearly visualized. CONCLUSIONS: The in vitro and in vivo targeting properties of 123I- and 131I-labeled mAb 14C5 are promising and could provide a new antibody-based agent for radioimmunodetection and radioimmunotherapy of patients bearing antigen 14C5-expressing tumors. PMID- 16243798 TI - Nuclear estrogen receptor beta in lung cancer: expression and survival differences by sex. AB - PURPOSE: A role for estrogens in determining lung cancer risk and prognosis is suggested by reported sex differences in susceptibility and survival. Archival lung tissue was evaluated for the presence of nuclear estrogen receptor (ER) alpha and ER-beta and the relationship between ER status, subject characteristics, and survival. EXPERIMENTAL DESIGN: Paraffin-embedded lung tumor samples were obtained from 214 women and 64 men from two population-based, case control studies as were 10 normal lung autopsy samples from patients without cancer. Nuclear ER-alpha and ER-beta expression was determined by immunohistochemistry. Logistic regression was used to identify factors associated with ER positivity and Cox proportional hazards models were used to measure survival differences by ER status. RESULTS: Neither tumor (0 of 94) nor normal (0 of 10) lung tissue stained positive for ER-alpha. Nuclear ER-beta positivity was present in 61% of tumor tissue samples (170 of 278; 70.3% in men and 58.3% in women) and 20% of normal tissue samples (2 of 10; P = 0.01). In multivariate analyses, females were 46% less likely to have ER-beta-positive tumors than males (odds ratio, 0.54; 95% confidence interval, 0.27-1.08). This relationship was stronger and statistically significant in adenocarcinomas (odds ratio, 0.40; 95% confidence interval, 0.18-0.89). Women with ER-beta-positive tumors had a nonsignificant 73% (P = 0.1) increase in mortality, whereas men with ER-beta positive tumors had a significant 55% (P = 0.04) reduction in mortality compared with those with ER-beta-negative tumors. CONCLUSIONS: This study suggests differential expression by sex and influence on survival in men of nuclear ER beta in lung cancer, particularly in adenocarcinomas. PMID- 16243800 TI - Total soluble HLA class I and soluble HLA-G in multiple myeloma and monoclonal gammopathy of undetermined significance. AB - Serum beta2-microglobulin, the light chain of the HLA class I molecular complex, remains one of the best survival prognostic factors in multiple myeloma, but other HLA class I molecules might be of interest in monoclonal gammopathies. In this study, we evaluate total soluble HLA class I (HLA-Is) and soluble HLA-G (HLA Gs) in 103 patients with newly diagnosed multiple myeloma, 30 patients with monoclonal gammopathy of undetermined significance (MGUS), and 30 healthy subjects, studying their prognostic value in multiple myeloma. In multiple myeloma patients, HLA-Is and HLA-Gs median values were 0.8 microg/mL and 28 ng/mL, respectively. Median HLA-Is concentration was higher in stage II and III multiple myeloma patients than in stage I multiple myeloma, MGUS, and control patients. Median HLA-Gs was significantly lower in healthy controls than in MGUS and multiple myeloma patients. A high level of HLA-Is (> or =2.1 microg/mL) was predictive of short survival (P = 0.017). For each given level of beta2 microglobulin, the relative risk of death was higher for patients with HLA-Is > or = 2.1 microg/mL than in patients with a lower level (P = 0.047). HLA-Gs, a marker of monoclonal gammopathy, was of no prognostic value, but the addition of HLA-Is to beta2-microglobulin produced an efficient prognostic score (P < 0.0001). HLA-Is is a new marker of multiple myeloma tumor load and provides additional survival prognostic information to beta2-microglobulin. PMID- 16243801 TI - Differential features of colorectal cancers fulfilling Amsterdam criteria without involvement of the mutator pathway. AB - PURPOSE: Hereditary nonpolyposis colorectal cancer (HNPCC) is the commonest form of inherited colorectal cancer. Whereas it has been known that mismatch repair gene mutations are the underlying cause of HNPCC, an undetermined number of patients do not have these alterations. The main objectives of this study were to assess the relevance of clinically defined HNPCC patients without characteristic mutator pathway alterations and to identify their specific features. EXPERIMENTAL DESIGN: This was a prospective, population-based, cohort that included 1,309 newly diagnosed colorectal cancer patients. Demographic, clinical, pathologic data and tumor DNA from probands as well as a detailed family history were collected. Microsatellite analysis and MLH1, MSH2, and MSH6 immunohistochemistry were done. Germ line MLH1 and MSH2 mutational analysis was done in all patients with evidence of MMR alterations. RESULTS: Twenty-five patients (1.9%) fulfilled Amsterdam criteria of HNPCC but 15 (60%) of them did not have microsatellite instability and showed normal expression of MMR proteins. These patients presented mostly left-sided tumors without lymphocytic infiltrate; they were older, had fewer family members affected with colorectal or endometrial cancers, and more often fulfilled Amsterdam II criteria than HNPCC patients with microsatellite instability. Like unstable HNPCC patients, this group without mutator pathway alterations had a significant percentage of synchronous and metachronous adenomatous polyps and cancers. CONCLUSIONS: We define an important group of HNPCC families with specific features, no evidence of mismatch repair deficiency, and an autosomal dominant trait with a lesser penetrance than HNPCC with deficiency. PMID- 16243802 TI - How ADAM-9 and ADAM-11 differentially from estrogen receptor predict response to tamoxifen treatment in patients with recurrent breast cancer: a retrospective study. AB - PURPOSE: To evaluate the predictive value of the disintegrin and metalloproteinases, ADAM-9, ADAM-10, ADAM-11, and ADAM-12, and of the matrix metalloproteinases, MMP-2 and MMP-9, in patients with recurrent breast cancer treated with tamoxifen. EXPERIMENTAL DESIGN: A retrospective study was done on 259 frozen specimens of estrogen receptor-positive primary breast carcinomas from patients who developed recurrent disease and were treated with tamoxifen as the first line of therapy. The expression levels of the biological factors were assessed by real-time quantitative reverse transcriptase PCR. RESULTS: Using log transformed continuous variables, increasing levels of ADAM-9 [odds ratio (OR) = 1.41; P = 0.015] and decreasing levels of MMP-9 (OR, 0.81; P = 0.035) predicted favorable disease control independent from the traditional predictive factors. Furthermore, when tumors were dichotomized at the median level of 70% tumor cell nuclei, our univariate analysis showed particularly strong results for the group of 153 patients with primary tumors containing 30% or more stromal cells. Although estrogen receptor levels lost their predictive power for this group of patients, high levels of ADAM-9 (OR, 1.59; P = 0.007) and ADAM-11 (OR, 1.65; P = 0.001) were significantly associated with a higher efficacy of tamoxifen therapy. CONCLUSIONS: Our results show that especially for primary tumors containing stromal elements, the assessment of mRNA expression levels of ADAM-9 and ADAM-11 could be useful to identify patients with recurrent breast cancer who are likely to benefit or fail from tamoxifen therapy. PMID- 16243803 TI - Increased intratumor Valpha24-positive natural killer T cells: a prognostic factor for primary colorectal carcinomas. AB - PURPOSE: Human invariant natural killer T (NKT) cells are novel, distinct lymphocyte populations with a restricted T-cell receptor repertoire (Valpha24 Vbeta11). They play a pivotal role in immunoregulation and in antitumor activities. This study focused on Valpha24+ NKT cells in colorectal carcinomas and their clinicopathologic significance. EXPERIMENTAL DESIGN: Valpha24+ NKT-cell infiltration immunohistochemistry was studied in a total of 103 colorectal carcinoma cases. The degree of NKT-cell infiltration in tumors was evaluated as low (<7 NKT cells/5 HPF) or high (> or =7 NKT cells/5 HPF). The correlation between the degree of infiltrated Valpha24+ NKT cells and clinicopathologic variables was studied statistically. RESULTS: A small number of Valpha24+ NKT cells were found in the normal colorectal mucosa (2.6 +/- 3.7 cells/5 HPF); however, their number increased remarkably in colorectal carcinomas (15.2 +/- 16.3 cells/5 HPF; P = 0.0003) and a majority showed phenotype of activation. Higher NKT-cell infiltration was more frequent in women than in men (P = 0.034) and correlated with fewer lymph node metastases (P = 0.042). Patients with high NKT-cell infiltration showed higher overall (P = 0.018) as well as disease-free (P = 0.0006) survival rates. Intratumor NKT-cell infiltration was an independent prognostic factor for the overall (P = 0.033) and disease-free (P = 0.0064) survival rates. CONCLUSIONS: Increased infiltration of Valpha24+ NKT cells was observed in colorectal carcinomas. Higher Valpha24+ NKT-cell infiltration in colorectal carcinomas was an independent prognostic factor for favorable prognosis. PMID- 16243804 TI - p53 Codon 72 polymorphism predicts the pathologic response to neoadjuvant chemotherapy in patients with breast cancer. AB - PURPOSE: Recent studies have highlighted that the p53 codon 72 polymorphism plays a crucial role in modulating wild-type p53 apoptotic capacity, and as such may influence the response to chemotherapy. Thus, the purpose of this study was to investigate whether the p53 codon 72 polymorphism might influence pathologic response to neoadjuvant chemotherapy in primary breast cancer. EXPERIMENTAL DESIGN: One hundred and ten operable breast cancer patients received anthracycline-based neoadjuvant chemotherapy and p53 codon 72 polymorphism status was analyzed by PCR-RFLP. RESULTS: The distribution of initial clinical stage, tumor size, estrogen receptor or progesterone receptor status, menopausal status, or erbB2 expression was not significantly different among the polymorphic variants. However, we found that only 13% (3 of 23) of patients with the Pro/Pro variant had a good pathologic response, defined as a complete pathologic response or minimal residual disease. In comparison, 40% (22 of 55) or 37.5% (12 of 32) of patients with the Pro/Arg or Arg/Arg variant had a good pathologic response (P = 0.019). Moreover, patients with the Pro/Pro variant were more likely to have a positive axillary lymph node status than those with the Pro/Arg or Arg/Arg variant (P = 0.007). Furthermore, in multivariate analysis, p53 codon 72 polymorphism was found to be a strong predictor of pathologic response (odds ratio 6.7, 95% confidence interval, 1.4-31.2; P = 0.016). CONCLUSION: Our study indicates that breast cancer patients with the Pro/Pro variant may be less sensitive to anthracycline-based treatment than those with the Pro/Arg or Arg/Arg variant and suggests that analysis of p53 codon 72 polymorphism may provide a simple predictive marker for selecting the right breast cancer patients to anthracycline-based neoadjuvant chemotherapy in clinical setting. PMID- 16243805 TI - Expressions of nuclear factor kappaB, inducible nitric oxide synthase, and vascular endothelial growth factor in adenoid cystic carcinoma of salivary glands: correlations with the angiogenesis and clinical outcome. AB - OBJECTIVE: To evaluate the expressions of nuclear factor kappaB (NF-kappaB p65), inducible nitric oxide synthase enzyme (iNOS), and vascular endothelial growth factor (VEGF) in relation to angiogenesis (microvessel density, MVD) and clinical outcomes in adenoid cystic carcinoma (ACC) of salivary glands. METHODS: Immunohistochemical staining was used to quantify the protein expression levels of NF-kappaB p65, iNOS, and VEGF in 80 surgically resected ACCs and 20 normal salivary tissues. In all cases of ACCs, MVD was evaluated by counting CD34 reactive endothelial cells or endothelial cell clusters. RESULTS: The nuclear localization of NF-kappaB p65 was only detected in ACC cells. Both iNOS and VEGF staining activities in ACCs were more significant than those in normal gland tissues (P < 0.01). MVD had significant correlations with NF-kappaB p65, iNOS, and VEGF expressions (P < 0.01). In three histologic types of ACCs, the NF kappaB, iNOS, VEGF expressions, and MVD were significantly higher in solid type than in cribriform and tubular types (P < 0.01). The NF-kappaB, iNOS, VEGF expressions, and MVD were significantly correlated with clinical stage, tumor size, vascular invasion, recurrence, and metastasis (P < 0.05). Multivariate analysis showed NF-kappaB, iNOS and VEGF expression, MVD, solid histotype, and perineural invasion had an independent prognostic effect on overall survival. CONCLUSION: The expressions of NF-kappaB p65, iNOS, and VEGF were related with MVD. Clinical outcomes raised the possibility that the overexpression of these cytokines might contribute to tumor angiogenesis and have prognostic value in ACCs. PMID- 16243806 TI - Lymphangiogenesis correlates with lymph node metastasis, prognosis, and angiogenic phenotype in human non-small cell lung cancer. AB - PURPOSE: Recent experimental studies have revealed that lymphangiogenesis plays an important role in cancer progression, but its clinical significance in the case of non-small cell lung cancer (NSCLC) remains unclear. Our aim was to assess the lymphangiogenesis of human NSCLC, and to correlate this with angiogenic phenotype (angiogenic versus nonangiogenic growth pattern) and clinical behavior. EXPERIMENTAL DESIGN: One hundred and three patients with NSCLC and complete follow-up information were included. Tumor samples were immunostained for vascular endothelial growth factor-C (VEGF-C), the lymphatic endothelial markers, LYVE-1 and D2-40/Podoplanin, and the panvascular marker, CD31. Lymphatic vessel density (LVD) and perimeters were evaluated within the tumor and peritumorally. RESULTS: LVDs at the tumor periphery were significantly higher in lymph node metastatic tumors (P < 0.005) and high LVDs correlated with poor overall survival (P < 0.001). However, this tendency proved to be significant only in the angiogenic tumor group (P < 0.001). Although 68% of the patients with nonangiogenic tumors had lymph node metastasis (P = 0.0048 versus angiogenic tumors), in the patient group with nonangiogenic NSCLCs, there was no information from the LVDs in any investigated tumor area (P > 0.05). In contrast to angiogenic tumors, which had actively sprouting lymphatics in all of the investigated tumor areas, nonangiogenic tumors showed no Ki67 staining intratumorally. CONCLUSIONS: Our results reveal tumor lymphangiogenesis as a novel prognostic indicator for the risk of lymph node metastasis in NSCLC. Moreover, it also provides the first evidence that nonangiogenic NSCLCs mainly co opt host tissue lymphatics during their growth, in contrast to most of the angiogenic tumors, which expand with concomitant lymphangiogenesis. PMID- 16243807 TI - The up-regulation of Y-box binding proteins (DNA binding protein A and Y-box binding protein-1) as prognostic markers of hepatocellular carcinoma. AB - PURPOSE: The development of hepatocellular carcinoma is associated with the chronic inflammation of the liver caused by various factors such as hepatitis B or C virus infection. Previously, we reported DNA binding protein A (dbpA) as a candidate molecule that can accelerate inflammation-induced hepatocarcinogenesis. DbpA belongs to the Y-box binding protein family, and Y-box binding protein-1 (YB 1), the prototype member of this family, is reported to be a prognostic marker of malignant diseases other than hepatocellular carcinoma. The purpose of this study is to examine the significance of the expression of dbpA or of the T-to-G transversion in the dbpA promoter region, which enhances the promoter activity in vitro, for the progression of hepatocellular carcinoma. EXPERIMENTAL DESIGN: We studied the expression of dbpA (as well as of YB-1) in 82 formalin-fixed hepatocellular carcinoma tissues by immunohistochemistry and determined the sequence of the dbpA promoter region in 42 frozen hepatocellular carcinoma tissues. We examined the relationship between these findings and the clinicopathologic factors of hepatocellular carcinoma patients. RESULTS: DbpA expression was associated with the advanced stages of hepatocellular carcinoma, and the cases with the nuclear dbpA expression had a poor prognosis. DbpA contributed more significantly to this association than YB-1. Furthermore, the T to-G transversion in the dbpA promoter region was related to the nuclear localization of dbpA. CONCLUSION: DbpA was a more significant prognostic marker of hepatocellular carcinoma than YB-1. The T-to-G transversion in the dbpA promoter region was suggested to be a predisposing factor for the progression of hepatocellular carcinoma. PMID- 16243808 TI - Cyclooxygenase-2 is an independent prognostic factor in gastric cancer and its expression is regulated by the messenger RNA stability factor HuR. AB - PURPOSE: Cyclooxygenase-2 (COX-2) promotes carcinogenesis and its expression associates with clinicopathologic characteristics in gastric cancer. HuR is an mRNA binding protein that controls the stability of certain transcripts including COX-2. We evaluated the prognostic significance of COX-2 and HuR expressions in gastric cancer and whether there exists a link between HuR and COX-2 expressions. EXPERIMENTAL DESIGN: The study included 342 consecutive patients with histologically confirmed gastric adenocarcinoma, of whom 321 patients had tissue specimens available for COX-2 and 316 for HuR immunohistochemistry. Specimens were stained by COX-2- and HuR-specific monoclonal antibodies and scored by two independent observers. Correlation to clinical data and survival was assessed. TMK-1 gastric adenocarcinoma cells were treated with small interfering RNA against HuR and expressions of HuR and COX-2 were detected by immunofluorescence and Western blot analysis. RESULTS: Patients with low COX-2 expression had a cumulative 5-year survival of 53% and those with high COX-2 expression had 16% (P < 0.0001). In multivariate analysis, COX-2 was an independent prognostic factor (P = 0.003). Cytoplasmic HuR expression was associated with high COX-2 expression (P < 0.0001) and with reduced survival (P = 0.004) whereas nuclear positivity for HuR was not. When TMK-1 cells were treated with HuR small interfering RNA, expressions of HuR and COX-2 were reduced. CONCLUSIONS: High COX-2 is an independent prognostic factor in gastric cancer. Cytoplasmic expression of HuR associates with high COX-2 expression and with reduced survival, and tissue culture experiments show that HuR can regulate expression of COX-2 in gastric cancer cells. PMID- 16243809 TI - Profiling cytochrome P450 expression in ovarian cancer: identification of prognostic markers. AB - PURPOSE: The cytochromes P450 are a multigene family of enzymes with a central role in the oxidative metabolism of a wide range of xenobiotics, including anticancer drugs and biologically active endogenous compounds. The purpose of this study was to define the cytochrome P450 profile of ovarian cancer and identify novel therapeutic targets and establish the prognostic significance of expression of individual cytochrome P450s in this type of cancer. EXPERIMENTAL DESIGN: Immunohistochemistry for a panel of 23 cytochrome P450s and cytochrome P450 reductase was done on an ovarian cancer tissue microarray consisting of 99 primary epithelial ovarian cancers, 22 peritoneal metastasis, and 13 normal ovarian samples. The intensity of immunoreactivity in each sample was established by light microscopy. RESULTS: In primary ovarian cancer, several P450s (CYP1B1, CYP2A/2B, CYP2F1, CYP2R1, CYP2U1, CYP3A5, CYP3A7, CYP3A43, CYP4Z1, CYP26A1, and CYP51) were present at a significantly higher level of intensity compared with normal ovary. P450 expression was also detected in ovarian cancer metastasis and CYP2S1 and P450 reductase both showed significantly increased expression in metastasis compared with primary ovarian cancer. The presence of low/negative CYP2A/2B (log rank = 7.06, P = 0.008) or positive CYP4Z1 (log rank = 6.19, P = 0.01) immunoreactivity in primary ovarian cancer were each associated with poor prognosis. Both CYP2A/2B and CYP4Z1 were also independent markers of prognosis. CONCLUSIONS: The expression profile of individual P450s has been established in ovarian cancer. Several P450s show increased expression in ovarian cancer and this provides the basis for developing P450-based therapeutics in ovarian cancer. Expression of CYP2A/2B or CYP4Z1 in primary ovarian cancer were independent markers of prognosis. PMID- 16243810 TI - Hypermethylation of 18S and 28S ribosomal DNAs predicts progression-free survival in patients with ovarian cancer. AB - PURPOSE: Repetitive ribosomal DNA (rDNA) genes are GC-rich clusters in the human genome. The aim of the study was to determine the methylation status of two rDNA subunits, the 18S and 28S genes, in ovarian tumors and to correlate methylation levels with clinicopathologic features in a cohort of ovarian cancer patients. EXPERIMENTAL DESIGN: 18S and 28S rDNA methylation was examined by quantitative methylation-specific PCR in 74 late-stage ovarian cancers, 9 histologically uninvolved, and 11 normal ovarian surface epithelial samples. In addition, methylation and gene expression levels of 18S and 28S rDNAs in two ovarian cancer cell lines were examined by reverse transcription-PCR before and after treatment with the demethylating drug 5'-aza-2'-deoxycytidine. RESULTS: The methylation level (amount of methylated rDNA/beta-actin) of 18S and 28S rDNAs was significantly higher (P < 0.05) in tumors than in normal ovarian surface epithelial samples. Methylation of 18S and 28S rDNA was highly correlated (R2= 0.842). Multivariate analysis by Cox regression found that rDNA hypermethylation [hazard ratio (HR), 0.25; P < 0.01], but not age (HR, 1.29; P = 0.291) and stage (HR, 1.09; P = 0.709), was independently associated with longer progression-free survival. In ovarian cancer cell lines, methylation levels of rDNA correlated with gene down-regulation and 5'-aza-2'-deoxycytidine treatment resulted in a moderate increase in 18S and 28S rDNA gene expressions. CONCLUSION: This is the first report of rDNA hypermethylation in ovarian tumors. Furthermore, rDNA methylation levels were higher in patients with long progression-free survival versus patients with short survival. Thus, rDNA methylation as a prognostic marker in ovarian cancer warrants further investigation. PMID- 16243811 TI - 14-3-3sigma in endometrial cancer--a possible prognostic marker in early-stage cancer. AB - PURPOSE: We examined expression of 14-3-3sigma, a regulator of cell proliferation, and evaluated its clinical significance in endometrioid endometrial carcinoma. EXPERIMENTAL DESIGN: One hundred three endometrioid endometrial adenocarcinoma cases were examined using immunohistochemistry with archival specimens. We correlated this finding with various clinicopathologic variables, including the status of estrogen receptor, progesterone receptor, and MIB-1 (Ki-57). RESULTS: 14-3-3sigma Immunoreactivity was detected in 78 of 103 (75.3%) of carcinoma cases. No statistically significant correlation was detected between status of 14-3-3sigma and any of clinicopathologic variables examined. There was, however, a statistically significant correlation between loss of 14-3 3sigma expression and adverse clinical outcome of the patients (P = 0.0007). In the early stages of cancer (stages I and II), 14-3-3sigma immunoreactivity was absent in 5 of 10 (50.0%) patients who showed recurrence during follow-up, whereas its absence was detected in only 13 of 68 (19.1%) disease-free patients in the same period. In addition, 14-3-3sigma immunoreactivity was absent in 4 of 5 (80.0%) patients who died, whereas its absence was detected in only 14 of 73 (19.2%) patients who had lived during the same period. Patients whose tumors were negative for 14-3-3sigma were at much greater risk to develop recurrent and/or mortal disease (P = 0.0372 and 0.0067). In multivariate analysis using the Cox proportional hazards model, absence of 14-3-3sigma turned out to be statistically independent risk factor in disease-free survival and overall survival even in patients with early-stage disease (P = 0.0321 and 0.0191). CONCLUSIONS: Results of our study showed that loss or absence of 14-3-3sigma determined by immunohistochemistry may be an important tool to identify endometrial carcinoma cases at high risk of recurrence and/or death, who are otherwise not detected by current clinical and pathologic evaluation, especially in the early stages of the disease. In addition, results of 14-3-3sigma immunohistochemistry in the early stage of endometrial carcinoma could contribute to planning postoperative follow up and adjuvant therapy. PMID- 16243812 TI - Reduction of Raf-1 kinase inhibitor protein expression correlates with breast cancer metastasis. AB - PURPOSE: Raf-1 kinase inhibitor protein (RKIP) was originally identified as the first physiologic inhibitor of the Raf/mitogen-activated protein kinase kinase/extracellular signal-regulated kinase (ERK) pathway. This pathway regulates fundamental cellular functions, including those that are subverted in cancer cells, such as proliferation, transformation, survival, and metastasis. Recently, RKIP has been recognized as a strong candidate for a metastasis suppressor gene in cell and animal model systems. Therefore, we investigated whether RKIP expression is altered in clinical specimens of human primary breast cancers and their lymph node metastases. EXPERIMENTAL DESIGN: Paraffin-embedded tumor samples from 103 breast cancer patients were examined immunohistochemically for the expression of RKIP, activated ERK, and apoptosis. The specificity of the antibodies used was validated by competition experiments with purified recombinant RKIP protein. RESULTS: RKIP expression was high in breast duct epithelia and retained to varying degrees in primary breast tumors. However, in lymph node metastases, RKIP expression was highly significantly reduced or lost (P = 0.000003). No significant correlations were observed between RKIP expression and histologic type, tumor differentiation grade, size, or estrogen receptor status. CONCLUSION: This is the first study of RKIP expression in a large clinical cohort. It confirms the results of cell culture and animal studies, suggesting that in human breast cancer, RKIP is a metastasis suppressor gene whose expression must be down-regulated for metastases to develop. RKIP expression is independent of other markers for breast cancer progression and prognosis. PMID- 16243813 TI - Effect of common CYP3A4 and CYP3A5 variants on the pharmacokinetics of the cytochrome P450 3A phenotyping probe midazolam in cancer patients. AB - PURPOSE: To evaluate the effect of naturally occurring variants in genes encoding the cytochrome P450 (CYP) isoforms CYP3A4 and CYP3A5 in patients with cancer receiving midazolam as a phenotyping probe. EXPERIMENTAL DESIGN: Five variants in CYP3A4 and CYP3A5 were evaluated in 58 patients (21 women and 37 men) receiving a short i.v. bolus of midazolam (dose, 0.0145 or 0.025 mg/kg). Midazolam concentrations in plasma were determined using liquid chromatography-mass spectrometry, and pharmacokinetic variables were calculated using noncompartmental analysis. Genomic DNA was characterized for the variants by PCR RFLP, and all genotypes were confirmed by direct nucleotide sequencing. RESULTS: The mean clearance of midazolam was 24.4 +/- 9.12 L/h, and phenotypic CYP3A activity varied about 4-fold in this population (range, 10.8-44.3 L/h). There were six carriers of the CYP3A4*1B allele (allele frequency, 0.061). No variant alleles for CYP3A4*17, CYP3A4*18A, or CYP3A5*6 were identified. Forty-eight of the 58 patients were homozygous variant for CYP3A5*3C, eight were heterozygous, and two were homozygous wild type (allele frequency, 0.897). No associations were noted between any of the studied genotypes and the phenotypic measures (P > or = 0.16). Likewise, a common variant in exon 26 in the gene encoding P-glycoprotein [i.e., ABCB1 (MDR1) 3435C>T] that was previously reported to be linked to CYP3A4 mRNA levels was unrelated to any of the studied phenotypic measures (P > or = 0.49). CONCLUSIONS: The studied genetic variants in CYP3A4 and CYP3A5 are unlikely to have an important functional significance to phenotypic CYP3A activity in patients with cancer. PMID- 16243814 TI - Apurinic/apyrimidinic endonuclease activity is associated with response to radiation and chemotherapy in medulloblastoma and primitive neuroectodermal tumors. AB - PURPOSE: Apurinic/apyrimidinic endonuclease (Ap endo) is a key DNA repair activity that confers resistance to radiation- and alkylator-induced cytotoxic abasic sites in human cells. We assayed apurinic/apyrimidinic endonuclease activity in medulloblastomas and primitive neuroectodermal tumors (PNET) to establish correlates with tumor and patient characteristics and with response to adjuvant radiation plus multiagent chemotherapy. EXPERIMENTAL DESIGN: Ap endo activity was assayed in 52 medulloblastomas and 10 PNETs from patients 0.4 to 21 years old. Ape1/Ref-1, the predominant human Ap endo activity, was measured in 42 medulloblastomas by immunostaining. Cox proportional hazards regression models were used to analyze the association of activity with time to tumor progression (TTP). RESULTS: Tumor Ap endo activity varied 180-fold and was significantly associated with age and gender. Tumor Ape1/Ref-1 was detected almost exclusively in nuclei. In a multivariate model, with Ap endo activity entered as a continuous variable, the hazard ratio for progression after adjuvant treatment in 46 medulloblastomas and four PNETs increased by a factor of 1.073 for every 0.01 unit increase in activity (P < or = 0.001) and was independent of age and gender. Suppressing Ap endo activity in a human medulloblastoma cell line significantly increased sensitivity to 1,3-bis(2-chlororethyl)-1-nitrosourea and temozolomide, suggesting that the association of tumor activity with TTP reflected, at least in part, abasic site repair. CONCLUSIONS: Our data (a) suggest that Ap endo activity promotes resistance to radiation plus chemotherapy in medulloblastomas/PNETs, (b) provide a potential marker of treatment outcome, and (c) suggest clinical use of Ap endo inhibitors to overcome resistance. PMID- 16243815 TI - Modeling the pharmacodynamics of highly schedule-dependent agents: exemplified by cytarabine-based regimens in acute myeloid leukemia. AB - BACKGROUND: Many agents in antineoplastic chemotherapy are highly schedule dependent. Therefore, variables such as total dose and also the area under the curve (AUC) that are schedule insensitive are generally insufficient to adequately represent treatment strength. PURPOSE: To establish a descriptor of treatment strength that takes into account the differential contribution of plasma concentrations (C) and exposure times (T) towards the cytotoxic effect and to investigate whether such a pharmacodynamically weighed descriptor is better correlated to the clinical effect than conventional variables. PATIENTS AND METHODS: The paradigm "C(N) x T = constant" (for an isoeffect) incorporates a weighing factor N (concentration coefficient) into the conventional description of the AUC that quantitates the differential contribution of C and T towards the cytotoxic effect. N was to be numerically derived from a multitude of in vitro isoeffect analyses of the major agents in acute myeloid leukemia (AML) therapy from patient samples (n = 57). RESULTS: For cytarabine, N was 0.45, numerically expressing the substantially higher relevance of T versus C for its cytotoxic effect. In a meta-analysis of 49 study arms involving >10,000 patients, neither total dose, dose intensity, nor AUC was correlated to the clinical effect. However, when AUC was pharmacodynamically weighed (N-weighed AUC, N-AUC = C(0.45) x T), this new descriptor was highly significantly correlated to the clinical effect (P < 0.001). CONCLUSION: The N-AUC concept is able to characterize schedule-dependent agents and is the only descriptor of cytarabine treatment strength actually correlated to the clinical effect in AML. PMID- 16243816 TI - CD4 and CD8 T-lymphocyte apoptosis can predict radiation-induced late toxicity: a prospective study in 399 patients. AB - PURPOSE: Predicting late effects in patients treated with radiation therapy by assessing in vitro radiation-induced CD4 and CD8 T-lymphocyte apoptosis can be useful in individualizing treatment. EXPERIMENTAL DESIGN: In a prospective study, 399 curatively irradiated patients were tested using a rapid assay where fresh blood samples were in vitro irradiated with 8 Gy X-rays. Lymphocytes were collected and prepared for flow cytometric analysis. Apoptosis was assessed by associated condensation of DNA. The incidences of late toxicities were compared for CD4 and CD8 T-lymphocyte apoptoses using receiver-operating characteristic curves and cumulative incidence. RESULTS: No association was found between early toxicity and T-lymphocyte apoptosis. Grade 2 and 3 late toxicities were observed in 31% and 7% of patients, respectively. More radiation-induced T-lymphocyte apoptosis was significantly associated with less grade 2 and 3 late toxicity (Gray's test, P < 0.0001). CD8 (area under the curve = 0.83) was more sensitive and specific than CD4. No grade 3 late toxicity was observed for patients with CD4 and CD8 values greater than 15% and 24%, respectively. The 2-year cumulative incidence for grade 2 or 3 late toxicity was 70%, 32%, and 12% for patients with absolute change in CD8 T-lymphocyte apoptosis of < or =16, 16 to 24, and >24, respectively. CONCLUSIONS: Radiation-induced T-lymphocyte apoptosis can significantly predict differences in late toxicity between individuals. It could be used as a rapid screen for hypersensitive patients to radiotherapy. In future dose escalation studies, patients could be selected using the apoptosis assay. PMID- 16243817 TI - Gene expression profile associated with response to doxorubicin-based therapy in breast cancer. AB - PURPOSE: This study was designed to identify genes that could predict response to doxorubicin-based primary chemotherapy in breast cancer patients. EXPERIMENTAL DESIGN: Biopsy samples were obtained before primary treatment with doxorubicin and cyclophosphamide. RNA was extracted and amplified and gene expression was analyzed using cDNA microarrays. RESULTS: Response to chemotherapy was evaluated in 51 patients, and based on Response Evaluation Criteria in Solid Tumors guidelines, 42 patients, who presented at least a partial response (> or =30% reduction in tumor dimension), were classified as responsive. Gene profile of samples, divided into training set (n = 38) and independent validation set (n = 13), were at first analyzed against a cDNA microarray platform containing 692 genes. Unsupervised clustering could not separate responders from nonresponders. A classifier was identified comprising EMILIN1, FAM14B, and PBEF, which however could not correctly classify samples included in the validation set. Our next step was to analyze gene profile in a more comprehensive cDNA microarray platform, containing 4,608 open reading frame expressed sequence tags. Seven samples of the initial training set (all responder patients) could not be analyzed. Unsupervised clustering could correctly group all the resistant samples as well as at least 85% of the sensitive samples. Additionally, a classifier, including PRSS11, MTSS1, and CLPTM1, could correctly distinguish 95.4% of the 44 samples analyzed, with only two misclassifications, one sensitive sample and one resistant tumor. The robustness of this classifier is 2.5 greater than the first one. CONCLUSION: A trio of genes might potentially distinguish doxorubicin responsive from nonresponsive tumors, but further validation by a larger number of samples is still needed. PMID- 16243818 TI - Long-term follow-up of patients with malignant pleural mesothelioma receiving high-dose adenovirus herpes simplex thymidine kinase/ganciclovir suicide gene therapy. AB - PURPOSE: Delineation of the long-term follow-up data on a series of patients with malignant mesothelioma, who received a single intrapleural dose of a nonreplicative adenoviral (Ad) vector encoding the herpes simplex virus thymidine kinase "suicide gene" (Ad.HSVtk) in combination with systemic ganciclovir. EXPERIMENTAL DESIGN: This report focuses on the 21 patients receiving "high-dose" therapy, defined by an intrapleural dose of vector (> or =1.6 x 10(13) viral particles), where transgene-encoded tk protein was reliably identified on immunohistochemical staining. In 13 patients, the vector was deleted in the E1 and E3 regions of the Ad; in the other eight patients, the vector had deletions in the Ad genes E1 and E4. Safety, immunologic responses, transgene expression, and clinical responses were evaluated. RESULTS: Both the E1/E3-deleted vector and the E1/E4-deleted vector were well tolerated and safe, although production of the E1/E4 vector was more difficult. Posttreatment antibody responses against the tumors were consistently seen. Interestingly, we observed a number of clinical responses in our patients, including two long-term (>6.5 year) survivors, both of whom were treated with the E1/E4-deleted vector. CONCLUSIONS: Intrapleural Ad.HSVtk/ganciclovir is safe and well tolerated in mesothelioma patients and resulted in long-term durable responses in two patients. Given the limited amount of gene transfer observed, we postulate that Ad.HSVtk may have been effective due to induction of antitumor immune responses. We hypothesize that approaches aiming to augment the immune effects of Ad gene transfer (i.e., with the use of cytokines) may lead to increased numbers of therapeutic responses in otherwise untreatable pleural malignancies. PMID- 16243819 TI - Pilot trial of unlabeled and indium-111-labeled anti-prostate-specific membrane antigen antibody J591 for castrate metastatic prostate cancer. AB - BACKGROUND: Prostate-specific membrane antigen (PSMA) is a transmembrane glycoprotein primarily expressed on benign and malignant prostatic epithelial cells. J591 is an IgG1 monoclonal antibody that targets the external domain of the PSMA. The relationship among dose, safety, pharmacokinetics, and antibody dependent cellular cytotoxicity (ADCC) activation for unlabeled J591 has not been explored. PATIENTS AND METHODS: Patients with progressive metastatic prostate cancer despite androgen deprivation were eligible. Each patient received 10, 25, 50, and 100 mg of J591. Two milligrams of antibody, conjugated with the chelate 1,4,7,10-tetraazacyclododecane-N, N',N'',N'''-tetraacetic acid, were labeled with 5 mCi indium-111 (111In) as a tracer. One group of patients received unlabeled J591 before the labeled antibody; the other received both together. Toxicities, pharmacokinetic properties, biodistribution, ADCC induction, immunogenicity, and clinical antitumor effects were assessed. RESULTS: Fourteen patients were treated (seven in each group). Treatment was well tolerated. Biodistribution of 111In labeled J591 was comparable in both groups. The mean T1/2 was .96, 1.9, 2.75, and 3.47 days for the 10, 25, 50, and 100 mg doses, respectively. Selective targeting of 111In-labeled J591 to tumor was seen. Hepatic saturation occurred by the 25-mg dose. ADCC activity was proportional to dose. One patient showed a >50% prostate specific antigen decline. CONCLUSIONS: J591 is well tolerated in repetitive dose escalating administrations. The rate of serum clearance decreases with increasing antibody mass. ADCC activation is proportional to antibody mass. The optimal dose is 25 mg for radioimmunotherapy and 100 mg for immunotherapy. Phase II studies using J591 as a radioconjugate are under way. PMID- 16243820 TI - Association of pretherapeutic expression of chemotherapy-related genes with response to neoadjuvant chemotherapy in Barrett carcinoma. AB - PURPOSE: We analyzed pretherapeutic gene expression patterns of patients with locally advanced adenocarcinomas of the esophagus with regard to response to neoadjuvant chemotherapy. EXPERIMENTAL DESIGN: Pretherapeutic, paraffin-embedded, formalin-fixed endoscopic esophageal tumor biopsies of 38 patients with locally advanced esophageal adenocarcinomas (Barrett adenocarcinoma) were included. All patients underwent two cycles of cisplatin and 5-fluorouracil (5-FU) therapy with or without additional paclitaxel followed by abdominothoracal esophagectomy. RNA expression levels of 5-FU metabolism-associated genes thymidylate synthase, thymidine phosphorylase, dihydropyrimidine dehydrogenase, methylenetetrahydrofolate reductase, MAP7, and ELF3, of platinum- and taxane related genes caldesmon, ERCC1, ERCC4, HER-2/neu, and GADD45, and of multidrug resistance gene MRP1 were determined using real-time reverse transcriptase-PCR. Expression levels were correlated with response to chemotherapy, histopathologically assessed in surgically resected specimens. RESULTS: Responding patients showed significantly higher pretherapeutic expression levels of MTHFR (P = 0.012), caldesmon (P = 0.016), and MRP1 (P = 0.007). In addition, patients with high pretherapeutic MTHFR and MRP1 levels had a survival benefit after surgery (P = 0.013 and P = 0.015, respectively). Additionally, investigation of intratumoral heterogeneity of gene expression of relevant genes (MTHFR, caldesmon, HER-2/neu, ERCC4, and MRP1), verified in nine untreated Barrett adenocarcinomas by examination of five distinct tumor areas, revealed no significant heterogeneity in gene expression indicating that expression profiles obtained from biopsy material may yield a representative genetic expression profile of total tumor tissue. CONCLUSIONS: Our results indicate that determination of mRNA levels of few genes may be useful for the prediction of the success of neoadjuvant chemotherapy in individual cancer patients with locally advanced Barrett adenocarcinoma. PMID- 16243821 TI - Phase I clinical trial of a HER-2/neu peptide (E75) vaccine for the prevention of prostate-specific antigen recurrence in high-risk prostate cancer patients. AB - PURPOSE: The E75 peptide is an immunogenic peptide from the HER-2/neu protein that is substantially expressed in prostate cancer. We are conducting a clinical trial of an E75/granulocyte macrophage colony-stimulating factor vaccine to prevent post-prostatectomy prostate-specific antigen (PSA) recurrences in high risk prostate cancer (HRPC) patients. EXPERIMENTAL DESIGN: Prostate cancer patients at high risk for recurrence were prospectively evaluated and identified by the validated Center for Prostate Disease Research (CPDR)/CaPSURE high-risk equation. From these high-risk equation patients, 27 HER-2/neu-expressing prostate cancer patients were enrolled. HLA-A2+ patients (n = 17) were vaccinated, whereas HLA-A2- patients (n = 10) were followed as clinical controls. Local/systemic toxicities, immunologic responses, and time to recurrence were measured. RESULTS: This vaccine is safe with only minor toxicities observed. Additionally, the vaccine is immunogenic with all patients showing both in vivo and in vitro phenotypic and functional immune responses, although variable. HLA A2+ patients were found to have larger tumors, higher postoperative Gleason scores, and more high-risk CPDR scores than HLA-A2- patients. Despite these differences, disease-free survival was not different between the vaccinated HLA A2+ patients and the HLA-A2- controls at a median follow up of 23 months. Three of the four vaccinated patients that recurred had rising PSAs at the initiation of the trial. Ex vivo phenotypic assays were predictive of recurrences and correlated in general with functional assays. CONCLUSIONS: The E75 vaccine strategy is safe and effective in eliciting an immune response against the HER 2/neu protein in HRPC patients and may be useful as a preventive strategy against disease recurrence. Vaccination in response to a rising PSA may be too late. PMID- 16243822 TI - Epidermal growth factor receptor activity determines response of colorectal cancer cells to gefitinib alone and in combination with chemotherapy. AB - PURPOSE: Up to now, there have been no established predictive markers for response to epidermal growth factor receptor (EGFR/HER1/erbB1) inhibitors alone and in combination with chemotherapy in colorectal cancer. To identify markers that predict response to EGFR-based chemotherapy regimens, we analyzed the response of human colorectal cancer cell lines to the EGFR-tyrosine kinase inhibitor, gefitinib (Iressa, AstraZeneca, Wilmington, DE), as a single agent and in combination with oxaliplatin and 5-fluorouracil (5-FU). EXPERIMENTAL DESIGN: Cell viability was assessed using 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide and crystal violet cell viability assays and analyzed by ANOVA. Apoptosis was measured by flow cytometry, poly(ADP-ribose) polymerase, and caspase 3 cleavage. EGFR protein phosphorylation was detected by Western blotting. RESULTS: Cell lines displaying high constitutive EGFR phosphorylation (a surrogate marker for EGFR activity) were more sensitive to gefitinib. Furthermore, in cell lines exhibiting low constitutive EGFR phosphorylation, an antagonistic interaction between gefitinib and oxaliplatin was observed, whereas in cell lines with high basal EGFR phosphorylation, the interaction was synergistic. In addition, oxaliplatin treatment increased EGFR phosphorylation in those cell lines in which oxaliplatin and gefitinib were synergistic but down regulated EGFR phosphorylation in those lines in which oxaliplatin and gefitinib were antagonistic. In contrast to oxaliplatin, 5-FU treatment increased EGFR phosphorylation in all cell lines and this correlated with synergistic decreases in cell viability when 5-FU was combined with gefitinib. CONCLUSIONS: These results suggest that phospho-EGFR levels determine the sensitivity of colorectal cancer cells to gefitinib alone and that chemotherapy-mediated changes in phospho EGFR levels determine the nature of interaction between gefitinib and chemotherapy. PMID- 16243824 TI - Schedule-dependent drug effects of oral 5-iodo-2-pyrimidinone-2'-deoxyribose as an in vivo radiosensitizer in U251 human glioblastoma xenografts. AB - PURPOSE: 5-Iodo-2-pyrimidinone-2'-deoxyribose (IPdR) is an oral prodrug of 5-iodo 2'-deoxyuridine (IUdR), an in vitro/in vivo radiosensitizer. IPdR can be rapidly converted to IUdR by a hepatic aldehyde oxidase. Previously, we found that the enzymatic conversion of IPdR to IUdR could be transiently reduced using a once daily (q.d.) treatment schedule and this may affect IPdR-mediated tumor radiosensitization. The purpose of this study is to measure the effect of different drug dosing schedules on tumor radiosensitization and therapeutic index in human glioblastoma xenografts. EXPERIMENTAL DESIGN: Three different IPdR treatment schedules (thrice a day, t.i.d.; every other day, q.o.d.; every 3rd day, q.3.d.), compared with a q.d. schedule, were analyzed using athymic nude mice with human glioblastoma (U251) s.c. xenografts. Plasma pharmacokinetics, IUdR-DNA incorporation in tumor and normal proliferating tissues, tumor growth delay following irradiation, and body weight loss were used as end points. RESULTS: The t.i.d. schedule with the same total daily doses as the q.d. schedule (250, 500, or 1,000 mg/kg/d) improved the efficiency of IPdR conversion to IUdR. As a result, the percentage of IUdR-DNA incorporation was higher using the t.i.d. schedule in the tumor xenografts as well as in normal small intestine and bone marrow. Using a fixed dose (500 mg/kg) per administration, the q.o.d. and q.3.d. schedules also showed greater IPdR conversion than the q.d. schedule, related to a greater recovery of hepatic aldehyde oxidase activity prior to the next drug dosing. In the tumor regrowth assay, all IPdR treatment schedules showed significant increases of regrowth delays compared with the control without IPdR (q.o.d., 29.4 days; q.d., 29.7 days; t.i.d., 34.7 days; radiotherapy alone, 15.7 days). The t.i.d. schedule also showed a significantly enhanced tumor growth delay compared with the q.d. schedule. Additionally, the q.o.d. schedule resulted in a significant reduction in systemic toxicity. CONCLUSIONS: The t.i.d. and q.o.d. dosing schedules improved the efficiency of enzymatic activation of IPdR to IUdR during treatment and changed the extent of tumor radiosensitization and/or systemic toxicity compared with a q.d. dosing schedule. These dosing schedules will be considered for future clinical trials of IPdR-mediated human tumor radiosensitization. PMID- 16243823 TI - Curcumin suppresses the paclitaxel-induced nuclear factor-kappaB pathway in breast cancer cells and inhibits lung metastasis of human breast cancer in nude mice. AB - Currently, there is no effective therapy for metastatic breast cancer after surgery, radiation, and chemotherapy have been used against the primary tumor. Because curcumin suppresses nuclear factor-kappaB (NF-kappaB) activation and most chemotherapeutic agents activate NF-kappaB that mediates cell survival, proliferation, invasion, and metastasis, we hypothesized that curcumin would potentiate the effect of chemotherapy in advanced breast cancer and inhibit lung metastasis. We tested this hypothesis using paclitaxel (Taxol)-resistant breast cancer cells and a human breast cancer xenograft model. As examined by electrophoretic mobility gel shift assay, paclitaxel activated NF-kappaB in breast cancer cells and curcumin inhibited it; this inhibition was mediated through inhibition of IkappaBalpha kinase activation and IkappaBalpha phosphorylation and degradation. Curcumin also suppressed the paclitaxel-induced expression of antiapoptotic (XIAP, IAP-1, IAP-2, Bcl-2, and Bcl-xL), proliferative (cyclooxygenase 2, c-Myc, and cyclin D1), and metastatic proteins (vascular endothelial growth factor, matrix metalloproteinase-9, and intercellular adhesion molecule-1). It also enhanced apoptosis. In a human breast cancer xenograft model, dietary administration of curcumin significantly decreased the incidence of breast cancer metastasis to the lung and suppressed the expression of NF-kappaB, cyclooxygenase 2, and matrix metalloproteinase-9. Overall, our results indicate that curcumin, which is a pharmacologically safe compound, has a therapeutic potential in preventing breast cancer metastasis possibly through suppression of NF-kappaB and NF-kappaB-regulated gene products. PMID- 16243826 TI - MHC class I-related chain A conjugated to antitumor antibodies can sensitize tumor cells to specific lysis by natural killer cells. AB - PURPOSE: As a first step for the development of a new cancer immunotherapy strategy, we evaluated whether antibody-mediated coating by MHC class I-related chain A (MICA) could sensitize tumor cells to lysis by natural killer (NK) cells. EXPERIMENTAL DESIGN: Recombinant MICA (rMICA) was chemically conjugated to Fab' fragments from monoclonal antibodies specific for tumor-associated antigens, such as carcinoembryonic antigen, HER2, or CD20. RESULTS: Flow cytometry analysis showed an efficient coating of MICA-negative human cancer cell lines with the Fab rMICA conjugates. This was strictly dependent on the expression of the appropriate tumor-associated antigens in the target cells. Importantly, preincubation of the tumor cells with the appropriate Fab-rMICA conjugate resulted in NK cell-mediated tumor cell lysis. Antibody blocking of the NKG2D receptor in NK cells prevented conjugate-mediated tumor cell lysis. CONCLUSIONS: These results open the way to the development of immunotherapy strategies based on antibody-mediated targeting of MICA. PMID- 16243825 TI - In vivo therapeutic responses contingent on Fanconi anemia/BRCA2 status of the tumor. AB - PURPOSE: BRCA2, FANCC, and FANCG gene mutations are present in a subset of pancreatic cancer. Defects in these genes could lead to hypersensitivity to interstrand cross-linkers in vivo and a more optimal treatment of pancreatic cancer patients based on the genetic profile of the tumor. EXPERIMENTAL DESIGN: Two retrovirally complemented pancreatic cancer cell lines having defects in the Fanconi anemia pathway, PL11 (FANCC-mutated) and Hs766T (FANCG-mutated), as well as several parental pancreatic cancer cell lines with or without mutations in the Fanconi anemia/BRCA2 pathway, were assayed for in vitro and in vivo sensitivities to various chemotherapeutic agents. RESULTS: A distinct dichotomy of drug responses was observed. Fanconi anemia-defective cancer cells were hypersensitive to the cross-linking agents mitomycin C (MMC), cisplatin, chlorambucil, and melphalan but not to 5-fluorouracil, gemcitabine, doxorubicin, etoposide, vinblastine, or paclitaxel. Hypersensitivity to cross-linking agents was confirmed in vivo; FANCC-deficient xenografts of PL11 and BRCA2-deficient xenografts of CAPAN1 regressed on treatment with two different regimens of MMC whereas Fanconi anemia-proficient xenografts did not. The MMC response comprised cell cycle arrest, apoptosis, and necrosis. Xenografts of PL11 also regressed after a single dose of cyclophosphamide whereas xenografts of genetically complemented PL11(FANCC) did not. CONCLUSIONS: MMC or other cross-linking agents as a clinical therapy for pancreatic cancer patients with tumors harboring defects in the Fanconi anemia/BRCA2 pathway should be specifically investigated. PMID- 16243828 TI - Complete tumor response following intratumoral 32P BioSilicon on human hepatocellular and pancreatic carcinoma xenografts in nude mice. AB - PURPOSE: 32P BioSilicon is a new, implantable, radiological medical device that comprises particles of highly pure silicon encapsulating 32phosphorus (32P) for the treatment of unresectable solid tumors. Prior to administration, the device particles are suspended in a formulant which provides an even suspension of the intended dose for implantation. The primary objective of this animal trial study was to investigate the effects of intratumoral injection of 32)P BioSilicon on human hepatocellular (HepG2) and pancreatic carcinoma (2119) xenografts implanted in nude mice (BALB/c). A secondary objective was the histopathologic examination of the tumor foci and surrounding tissue during the study. METHODS: Cultured human carcinoma cells (HepG2 and 2119) were injected s.c. into the gluteal region of nude mice. When the implanted tumors were approximately 1 cm in diameter, 32P BioSilicon (0.5, 1.0, and 2.0 MBq) or formulant was injected into the tumors. Implanted tumor size was measured once a week for 10 weeks. At study termination, the tumor and surrounding normal tissue were collected and fixed in 10% formalin and processed for histopathologic analysis. RESULTS: 32P BioSilicon produced a reduction in HepG2 tumor volume when compared with formulant control, and complete response was observed among tumors in the 1.0 and 2.0 MBq treatment groups after week 8. There was also significant reduction in 2119 tumor volume in all treated groups, with the complete response rate of 67% in the 2.0 MBq group. CONCLUSION: 32P BioSilicon suppressed the growth of both human hepatocellular and pancreatic carcinoma xenografts implanted in nude mice and complete responses were also observed in tumors at higher radiation doses. PMID- 16243827 TI - Stable suppression of tumorigenicity by Pin1-targeted RNA interference in prostate cancer. AB - PURPOSE: The peptidyl-prolyl isomrase Pin1 plays a catalytic role in oncogenesis in solid cancers, including prostate cancer. In the present study, we sought to determine the potential of Pin1-targeted gene silencing in inhibiting cellular growth and tumorigenicity in prostate cancer. EXPERIMENTAL DESIGN: A retrovirus mediated RNA interference targeting Pin1 was expressed in PC3 and LNCaP cells, and cell growth and several transformed properties were investigated. RESULTS: The stable expression of Pin1-specific small interfering RNA constructs in PC3 and LNCaP cells significantly reduced cellular proliferation, colony formation, migration, and invasion but strongly enhanced the apoptotic response induced by serum depletion or treatment with anticancer agents. Furthermore, Pin1 depletion significantly suppressed tumorigenic potential in athymic mice, resulting in the inhibition of both tumor growth and angiogeneisis. CONCLUSIONS: These results strongly suggest that Pin1 plays an important role not only in tumorigenesis but also in the maintenance of the transformed phenotype in prostate cancer cells. Hence, Pin1 may serve as a promising therapeutic target, particularly for recurrent prostate tumors. PMID- 16243829 TI - Optimizing photodynamic therapy: in vivo pharmacokinetics of liposomal meta (tetrahydroxyphenyl)chlorin in feline squamous cell carcinoma. AB - PURPOSE: The aim of the present study was to optimize and simplify photodynamic therapy using a new liposomal formulation of the photosensitizer meta (tetrahydroxyphenyl)chlorin [m-THPC (Foscan); liposomal m-THPC (Fospeg)] and to reduce systemic reactions to the photosensitizer. EXPERIMENTAL DESIGN: To examine the pharmacokinetics of liposomal m-THPC, we determined tissue and plasma variables in feline patients with spontaneous squamous cell carcinoma. In vivo fluorescence intensity measurements of tumor and skin were done with a fiber spectrophotometer after i.v. injection of m-THPC or liposomal m-THPC in 10 cats. Blood samples, drawn at several time points after photosensitizer administration, were analyzed by high-performance liquid chromatography. RESULTS: None of the liposomal m-THPC-treated cats showed side effects during or after drug injection. Fluorescence intensities, fluorescence ratios (tumor fluorescence divided by skin fluorescence), and bioavailability in the tumor were 2 to 4 times higher with liposomal m-THPC compared with m-THPC. Liposomal m-THPC concentration in the tumor increased constantly to reach a maximum at 4 hours after injection. Plasma concentration and bioavailability were approximately 3 times higher with liposomal m-THPC compared with m-THPC measured at the time points of highest plasma concentration. The distribution half-life was shorter with liposomal m THPC, resulting in maximal tumor accumulation up to 5.5 times earlier. Maximal tumor accumulation and maximal fluorescence ratio with liposomal m-THPC occurred at the same time point, indicating maximal selectivity. In both groups, all cats responded to therapy. CONCLUSIONS: Liposomal m-THPC was well tolerated by all cats and seems to have superior pharmacokinetic properties compared with m-THPC. The efficacy of the drug warrants further study. PMID- 16243830 TI - Silencing of human phosphatidylethanolamine-binding protein 4 sensitizes breast cancer cells to tumor necrosis factor-alpha-induced apoptosis and cell growth arrest. AB - PURPOSE: The current therapeutic approach is not so effective in breast cancer patients. Alternative treatment protocols aimed at different targets need to be explored. We recently reported a novel phosphatidylethanolamine-binding protein, human phosphatidylethanolamine-binding protein 4 (hPEBP4), as an antiapoptotic molecule. The finding led us to explore a promising approach for breast cancer therapy via silencing the expression of hPEBP4. EXPERIMENTAL DESIGN: hPEBP4 expression in clinical breast specimens was examined by Tissue Microarrays. RNA interference was used to silence hPEBP4 expression in MCF-7 breast carcinoma cells and the effects on cell proliferation, cell cycle progression, apoptosis, as well as underlying mechanisms, were investigated. RESULTS: hPEBP4 was found to be expressed in up to 50% of breast cancers but in only <4% of normal breast tissues. Silencing of hPEBP4 potentiated tumor necrosis factor-alpha (TNF-alpha) induced apoptosis and cell cycle arrest in MCF-7 cells, which was due to the increased mitogen-activated protein kinase activation and the enhanced phosphatidylethanolamine externalization. Further investigation showed that silencing of hPEBP4 in MCF-7 cells promoted TNF-alpha-induced stability of p53, up-regulation of phospho-p53ser15, p21waf/cip, and Bax, and down-regulation of Bcl-2 and Bcl-xL, which were shown to depend on extracellular signal-regulated kinase 1/2 and c-jun NH2-terminal kinase activation by hPEBP4 silencing. Moreover, the increased proportion of cells in the G0-G1 phase of cell cycle was observed in hPEBP4-silenced MCF-7 cells on TNF-alpha treatment and the expression of cyclin A and cyclin E was down-regulated more significantly. CONCLUSIONS: The antiapoptotic effect and the preferential expression pattern in breast cancer tissues make hPEBP4 a new target for breast cancer therapy. Silencing of hPEBP4 expression may be a promising approach for the treatment of breast carcinoma. PMID- 16243831 TI - More efficient induction of HLA-A*0201-restricted and carcinoembryonic antigen (CEA)-specific CTL response by immunization with exosomes prepared from heat stressed CEA-positive tumor cells. AB - PURPOSE: Tumor-derived exosomes are proposed as a new type of cancer vaccine. Heat shock proteins are potent Th1 adjuvant, and heat stress can induce heat shock protein and MHC-I expression in tumor cells, leading to the increased immunogenicity of tumor cells. To improve the immunogenicity of exosomes as cancer vaccine, we prepared exosomes from heat-stressed carcinoembryonic antigen (CEA)-positive tumor cells (CEA+/HS-Exo) and tested the efficacy of these exosomes in the induction of CEA-specific antitumor immunity. EXPERIMENTAL DESIGN: First, we identified the composition of CEA+/HS-Exo and observed their effects on human dendritic cell maturation. Then, we evaluated their ability to induce a CEA-specific immune response in vivo in HLA-A2.1/Kb transgenic mice and CEA-specific CTL response in vitro in HLA-A*0201+ healthy donors and HLA A*0201+CEA+ cancer patients. RESULTS: CEA+/HS-Exo contained CEA and more heat shock protein 70 and MHC-I and significantly induced dendritic cell maturation. Immunization of HLA-A2.1/Kb transgenic mice with CEA+/HS-Exo was more efficient in priming a CEA-specific CTL, and the CTL showed antitumor effect when adoptively transferred to SW480-bearing nude mice. Moreover, in vitro incubation of lymphocytes from HLA-A*0201+ healthy donors and HLA-A*0201+CEA+ cancer patients with CEA+/HS-Exo-pulsed autologous dendritic cells induces HLA-A*0201 restricted and CEA-specific CTL response. CONCLUSIONS: Our results show that CEA+/HS-Exo has superior immunogenicity than CEA+/Exo in inducing CEA-specific CTL response and suggest that exosomes derived from heat-stressed tumor cells may be used as efficient vaccine for cancer immunotherapy. PMID- 16243832 TI - Kinetics of tempol for prevention of xerostomia following head and neck irradiation in a mouse model. AB - PURPOSE: Radiotherapy is commonly used to treat the majority of patients with head and neck cancers. Salivary glands in the radiation field are dramatically affected by this procedure. The purpose of this study was to examine pharmacokinetic characteristics of the stable nitroxide 4-hydroxy-2,2,6,6 tetramethylpiperidine-N-oxyl (tempol) with respect to radioprotection of the salivary glands. EXPERIMENTAL DESIGN: To evaluate the effect of different doses and times of administration, the heads of C3H mice were exposed to a single irradiation dose of 15 Gy, with i.p. tempol injection. To analyze other routes of administration, we injected 275 mg/kg tempol by an i.m., i.v., or s.c. route, 10 minutes before irradiation. We also tested whether oral administration of tempol in a topical form (either in a mouthwash or gel) provided any salivary gland protection. RESULTS: Tempol treatment (137.5 or 275 mg/kg, i.p., 10 minutes before irradiation) significantly reduced irradiation-induced salivary hypofunction (approximately 50-60%). I.v. or s.c. administration of tempol also showed significant radioprotection, whereas i.m. administration proved to be ineffective. Topical use of tempol, either as a mouthwash or gel, also was radioprotective. CONCLUSIONS: Our results strongly suggest that tempol is a promising candidate for clinical application to protect salivary glands in patients undergoing radiotherapy for head and neck cancers. PMID- 16243833 TI - Topotecan combination chemotherapy in two new rodent models of retinoblastoma. AB - Chemotherapy combined with laser therapy and cryotherapy has improved the ocular salvage rate for children with bilateral retinoblastoma. However, children with late-stage disease often experience recurrence shortly after treatment. To improve the vision salvage rate in advanced bilateral retinoblastoma, we have developed and characterized two new rodent models of retinoblastoma for screening chemotherapeutic drug combinations. The first model is an orthotopic xenograft model in which green fluorescent protein- or luciferase-labeled human retinoblastoma cells are injected into the eyes of newborn rats. The second model uses a replication-incompetent retrovirus (LIA-E(E1A)) encoding the E1A oncogene. Clonal, focal tumors arise from mouse retinal progenitor cells when LIA-E(E1A) is injected into the eyes of newborn p53-/- mice. Using these two models combined with pharmacokinetic studies and cell culture experiments, we have tested the efficacy of topotecan combined with carboplatin and of topotecan combined with vincristine for the treatment of retinoblastoma. The combination of topotecan and carboplatin most effectively halted retinoblastoma progression in our rodent models and was superior to the current triple drug therapy using vincristine, carboplatin, and etoposide. Vincristine had the lowest LC50 in culture but did not reduce tumor growth in our preclinical retinoblastoma models. Taken together, these data suggest that topotecan may be a suitable replacement for etoposide in combination chemotherapy for the treatment of retinoblastoma. PMID- 16243835 TI - S100A4, a mediator of metastasis. PMID- 16243834 TI - Estrogen receptor alpha-negative breast cancer tissues express significant levels of estrogen-independent transcription factors, ERbeta1 and ERbeta5: potential molecular targets for chemoprevention. AB - We have investigated the expression of two estrogen receptor beta (ERbeta) isoforms, ERbeta1 and ERbeta5, which activate gene transcription independent of estrogen or growth factors, in ERalpha-negative breast cancer tissues. We report here, for the first time, that ERalpha-negative tissues express significant levels of ERbeta1 and ERbeta5, and their expression levels are not different from levels in ERalpha positive tumors. However, significant differences exist between the two racial groups, African American and Caucasian, in that the patients from the former group express higher levels of ERbeta1 and ERbeta5 but not ERalpha. These two transcription factors could be potential molecular targets for designing chemopreventive drugs to treat ERalpha-negative breast cancers. PMID- 16243836 TI - Identification of the preprotein binding domain of SecA. AB - SecA, the preprotein translocase ATPase, has a helicase DEAD motor. To catalyze protein translocation, SecA possesses two additional flexible domains absent from other helicases. Here we demonstrate that one of these "specificity domains" is a preprotein binding domain (PBD). PBD is essential for viability and protein translocation. PBD mutations do not abrogate the basal enzymatic properties of SecA (nucleotide binding and hydrolysis), nor do they prevent SecA binding to the SecYEG protein conducting channel. However, SecA PBD mutants fail to load preproteins onto SecYEG, and their translocation ATPase activity does not become stimulated by preproteins. Bulb and Stem, the two sterically proximal PBD substructures, are physically separable and have distinct roles. Stem binds signal peptides, whereas the Bulb binds mature preprotein regions as short as 25 amino acids. Binding of signal or mature region peptides or full-length preproteins causes distinct conformational changes to PBD and to the DEAD motor. We propose that (a) PBD is a preprotein receptor and a physical bridge connecting bound preproteins to the DEAD motor, and (b) preproteins control the ATPase cycle via PBD. PMID- 16243837 TI - Calcium binding by the essential virulence factor BAD-1 of Blastomyces dermatitidis. AB - BAD-1 (Blastomyces adhesin 1), a 120-kDa protein of Blastomyces dermatitidis, functions as an adhesin, immune modulator, and essential virulence factor. Structurally, BAD-1 is composed of a short N-terminal region, a core of 30 tandem repeats critical for virulence, and a C-terminal epidermal growth factor domain that binds the protein to yeast cell surface chitin. Each of the 30 acidic residue-rich tandem repeats contains a sequence that resembles the calcium binding loop of the EF-hand domain found in many calcium-binding proteins. Here, we investigated the binding of calcium by BAD-1 and its biological significance. Yeast washed with double distilled H2O released surface-bound BAD-1, but EGTA washes were an order of magnitude more efficient, suggesting an interaction between BAD-1 and calcium. Immobilized BAD-1 was stained with ruthenium red dye, an indicator of calcium-binding proteins. In equilibrium dialysis, BAD-1 bound 45Ca2+ with an affinity of 0.41 x 10(-5) m and a capacity of 27 calcium/mol. Mass spectrometry confirmed this capacity. Elevated [Ca2+] diminished BAD-1 solubility. Upon deletion of its C-terminal epidermal growth factor-like domain, BAD-1 resisted aggregation by elevated [Ca2+] but retained its affinity and capacity for calcium. Removing 20 copies of the tandem repeat, however, sharply reduced the capacity of BAD-1 for calcium. Growth of the bad-1 null yeast was inhibited by 5 mm EGTA, and re-expression of BAD-1 in trans or the addition of exogenous purified BAD-1 restored growth. Thus, BAD-1 is a high capacity calcium binding protein. This property contributes to the structure and function of BAD 1, as well as to B. dermatitidis acquisition of calcium from the environment. PMID- 16243838 TI - Regulation of the interaction of inosine monophosphate dehydrogenase with mycophenolic Acid by GTP. AB - Inosine monophosphate dehydrogenase (IMPDH), a rate-limiting enzyme in the de novo synthesis of guanine nucleotides, is a major therapeutic target. A prototypic uncompetitive inhibitor of IMPDH, mycophenolic acid (MPA), is the active form of mycophenolate mofeteil (CellCept), a widely used immunosuppressive drug. We have found that MPA interacts with intracellular IMPDH in vivo to alter its mobility on SDS-polyacrylamide gels. MPA also induces a striking conformational change in IMPDH protein in intact cells, resulting in the formation of annular aggregates of protein with concomitant inhibition of IMPDH activity. These aggregates are not associated with any known intracellular organelles and are reversible by incubating cells with guanosine, which repletes intracellular GTP, or with GTPgammaS. GTP also restores IMPDH activity. Treatment of highly purified IMPDH with MPA also results in the formation of large aggregates of protein, a process that is both prevented and reversed by the addition of GTP. Finally, GTP binds to IMPDH at physiologic concentrations, induces the formation of linear arrays of tetrameric protein, and prevents the aggregation of protein induced by MPA. We conclude that intracellular GTP acts as an antagonist to MPA by directly binding to IMPDH and reversing the conformational changes in the protein. PMID- 16243839 TI - Perilipin targets a novel pool of lipid droplets for lipolytic attack by hormone sensitive lipase. AB - Adipocytes serve as the principal energy reservoir of the body; however, the subcellular organization of the machinery regulating lipid trafficking and metabolism is poorly understood. Mobilization of stored triglyceride is thought be controlled by interactions among intracellular lipases and proteins that coat lipid storage droplets. A major limitation of previous studies of hormone mediated lipolysis, however, is the use of cultured model adipocytes whose three dimensional architectures do not resemble those in real adipose tissue. To address this limitation, we investigated the intracellular targeting of perilipin, a major lipid coat protein, and hormone-sensitive lipase in three preparations that exhibit more appropriate morphologies: 3T3-L1 adipocytes grown in three-dimensional matrix, dissociated mature adipocytes from mouse adipose tissue, and adipocytes within intact fat pads. High resolution imaging of native and fluorescently tagged proteins indicate that: 1) perilipin preferentially targets a special class of peripheral lipid storage droplets, but not the major or central lipid storage droplets, 2) the peripheral droplets are the sites of attack by hormone-sensitive lipase, and 3) perilipin and hormone-sensitive lipase are continuously colocalized following lipolytic activation. These results indicate that in white adipose tissue, lipolysis takes place in a specialized subcellular domain that is distinct from the major lipid storage site and is defined by perilipin. PMID- 16243841 TI - Resistin-like molecule beta activates MAPKs, suppresses insulin signaling in hepatocytes, and induces diabetes, hyperlipidemia, and fatty liver in transgenic mice on a high fat diet. AB - Resistin and resistin-like molecules (RELMs) are a family of proteins reportedly related to insulin resistance and inflammation. Because the serum concentration and intestinal expression level of RELMbeta were elevated in insulin-resistant rodent models, in this study we investigated the effect of RELMbeta on insulin signaling and metabolism using transgenic mice and primary cultured hepatocytes. First, transgenic mice with hepatic RELMbeta overexpression were shown to exhibit significant hyperglycemia, hyperlipidemia, fatty liver, and pancreatic islet enlargement when fed a high fat diet. Hyperinsulinemic glucose clamp showed a decreased glucose infusion rate due to increased hepatic glucose production. In addition, the expression levels of IRS-1 and IRS-2 proteins as well as the degrees of insulin-induced phosphatidylinositol 3-kinase and Akt activations were attenuated in RELMbeta transgenic mice. Similar down-regulations of IRS-1 and IRS 2 proteins were observed in primary cultured hepatocytes chronically treated (for 24 h) with RELMbeta, suggesting the insulin resistance-inducing effect of RELMbeta to be direct. Furthermore, it was shown that RELMbeta acutely and markedly activates ERK and p38, while weakly activating JNK, in primary cultured hepatocytes. This increased basal p38 phosphorylation level was also observed in the livers of RELMbeta transgenic mice. In conclusion, RELMbeta, a gut-derived hormone, impairs insulin signaling probably via the activations of classic MAPKs, and increased expression of RELMbeta may be involved in the pathogenesis of glucose intolerance and hyperlipidemia in some insulin-resistant models. Thus, RELMbeta is a potentially useful marker for assessing insulin resistance and may also be a target for future novel anti-diabetic agents. PMID- 16243840 TI - Protein kinase B/Akt is a novel cysteine string protein kinase that regulates exocytosis release kinetics and quantal size. AB - Protein kinase B/Akt has been implicated in the insulin-dependent exocytosis of GLUT4-containing vesicles, and, more recently, insulin secretion. To determine if Akt also regulates insulin-independent exocytosis, we used adrenal chromaffin cells, a popular neuronal model. Akt1 was the predominant isoform expressed in chromaffin cells, although lower levels of Akt2 and Akt3 were also found. Secretory stimuli in both intact and permeabilized cells induced Akt phosphorylation on serine 473, and the time course of Ca2+-induced Akt phosphorylation was similar to that of exocytosis in permeabilized cells. To determine if Akt modulated exocytosis, we transfected chromaffin cells with Akt constructs and monitored catecholamine release by amperometry. Wild-type Akt had no effect on the overall number of exocytotic events, but slowed the kinetics of catecholamine release from individual vesicles, resulting in an increased quantal size. This effect was due to phosphorylation by Akt, because it was not seen in cells transfected with kinase-dead mutant Akt. As overexpression of cysteine string protein (CSP) results in a similar alteration in release kinetics and quantal size, we determined if CSP was an Akt substrate. In vitro 32P phosphorylation studies revealed that Akt phosphorylates CSP on serine 10. Using phospho-Ser10-specific antisera, we found that both transfected and endogenous cellular CSP is phosphorylated by Akt on this residue. Taken together, these findings reveal a novel role for Akt phosphorylation in regulating the late stages of exocytosis and suggest that this is achieved via the phosphorylation of CSP on serine 10. PMID- 16243842 TI - Synthetic Vpr protein activates activator protein-1, c-Jun N-terminal kinase, and NF-kappaB and stimulates HIV-1 transcription in promonocytic cells and primary macrophages. AB - The human immunodeficiency virus (HIV) Vpr protein plays a critical role in AIDS pathogenesis, especially by allowing viral replication within nondividing cells such as mononuclear phagocytes. Most of the data obtained so far have been in experiments with endogenous Vpr protein; therefore the effects of extracellular Vpr protein remain largely unknown. We used synthetic Vpr protein to activate nuclear transcription factors activator protein-1 (AP-1) and NF-kappaB in the promonocytic cell line U937 and in primary macrophages. Synthetic HIV-1 Vpr protein activated AP-1, c-Jun N-terminal kinase, and MKK7 in both U937 cells and primary macrophages. Synthetic Vpr activated NF-kappaB in primary macrophages and to a lesser extent in U937 cells. Because synthetic Vpr activated AP-1 and NF kappaB, which bind to the HIV-1 long terminal repeat, we investigated the effect of synthetic Vpr on HIV-1 replication. We observed that synthetic Vpr stimulated HIV-1 long terminal repeat in U937 cells and enhanced viral replication in chronically infected U1 promonocytic cells. Similarly, synthetic Vpr stimulated HIV-1 replication in acutely infected primary macrophages. Activation of transcription factors and enhancement of viral replication in U937 cells and primary macrophages were mediated by both the N-terminal and the C-terminal moieties of synthetic Vpr. Therefore, our results suggest that extracellular Vpr could fuel the progression of AIDS via stimulation of HIV-1 provirus present in such cellular reservoirs as mononuclear phagocytes in HIV-infected patients. PMID- 16243843 TI - Protamines, in the footsteps of linker histone evolution. PMID- 16243844 TI - PKCdelta mediates testosterone-induced increases in coronary smooth muscle Cav1.2. AB - Sex hormones have emerged as important modulators of cardiovascular physiology and pathophysiology. Our previous studies demonstrated that testosterone increases expression and activity of L-type, voltage-gated calcium channels (Cav1.2) in coronary arteries of males. The purpose of the present study was to determine whether testosterone (T) alters coronary protein kinase C delta (PKCdelta) expression and whether PKCdelta plays a role in coronary Cav1.2 expression. For in vitro studies, porcine right coronary arteries (RCA) and post confluent (passages 3-6) 5-day, serum-restricted coronary smooth muscle cell cultures (CSMC) were incubated in the presence and absence of T or dihydrotestosterone (10 and 100 nm) for 18 h at 37 degrees C in a humidified chamber. For sex and endogenous testosterone-dependent effects, RCA were obtained from intact males, castrated males, castrated males with T replacement, and intact females. In vitro T and dihydrotestosterone caused an approximately 2-3 fold increase in PKCdelta protein levels, approximately 1.5-2-fold increase in PKCdelta kinase activity, and localization of PKCdelta toward the plasma membrane and nuclear envelope. PKCdelta protein levels were higher in coronary arteries of intact males compared with intact females. Elimination of endogenous testosterone by castration reduced RCA PKCdelta protein levels, an effect partially (approximately 45%) reversed by exogenous T (castrated males with T replacement). In CSMC, PKC inhibition with either the general PKC inhibitor, cheylerythrine, or the putative PKCdelta inhibitor, rottlerin, completely inhibited the T-mediated increase in coronary Cav1.2 protein levels. Conversely, Go6976, a conventional PKC isoform inhibitor, failed to inhibit T-induced increases in coronary Cav1.2 protein levels. PKCdelta short interference RNA completely blocked T-induced increases in Cav1.2 protein levels in CSMC. These results demonstrate for the first time that 1) endogenous T is a primary modulator of coronary PKCdelta protein and activity in males and 2) T increases Cav1.2 protein expression in a PKCdelta-dependent manner. PMID- 16243845 TI - Rotenone model of Parkinson disease: multiple brain mitochondria dysfunctions after short term systemic rotenone intoxication. AB - Chronic infusion of rotenone (Rot) to Lewis rats reproduces many features of Parkinson disease. Rot (3 mg/kg/day) was infused subcutaneously to male Lewis rats for 6 days using Alzet minipumps. Control rats received the vehicle only. Presence of 0.1% bovine serum albumin during the isolation procedure completely removed rotenone bound to the mitochondria. Therefore all functional changes observed were aftereffects of rotenone toxicity in vivo. In Rot rat brain mitochondria (Rot-RBM) there was a 30-40% inhibition of respiration in State 3 and State 3U with Complex I (Co-I) substrates and succinate. Rot did not affect the State 4Deltapsi of RBM and rat liver mitochondria (RLM). However, Rot-RBM required two times less Ca2+ to initiate permeability transition (mPT). There was a 2-fold increase in O*2- or H2O2 generation in Rot-RBM oxidizing glutamate. Rot infusion affected RLM little. Our results show that in RBM, the major site of reactive oxygen species generation with glutamate or succinate is Co-I. We also found that Co-II generates substantial amounts of reactive oxygen species that increased 2-fold in the Rot-RBM. Our data suggest that the primary mechanism of the Rot toxic effect on RBM consists in a significant increase of O*2- generation that causes damage to Co-I and Co-II, presumably at the level of 4Fe-4S clusters. Decreased respiratory activity diminishes resistance of RBM to Ca2+ and thus increases probability of mPT and apoptotic cell death. We suggest that the damage to Co-I and Co-II shifts O*2- generation from the CoQ10 sites to more proximal sites, such as flavines, and makes it independent of the RBM functional state. PMID- 16243846 TI - The mechanism of membrane targeting of human sphingosine kinase 1. AB - Sphingosine 1-phosphate is a bioactive sphingolipid that regulates cell growth and suppresses programmed cell death. The biosynthesis of sphingosine 1-phosphate is catalyzed by sphingosine kinase (SK) but the mechanism by which the subcellular localization and activity of SK is regulated in response to various stimuli is not fully understood. To elucidate the origin and structural determinant of the specific subcellular localization of SK, we performed biophysical and cell studies of human SK1 (hSK1) and selected mutants. In vitro measurements showed that hSK1 selectively bound phosphatidylserine over other anionic phospholipids and strongly preferred the plasma membrane-mimicking membrane to other cellular membrane mimetics. Mutational analysis indicates that conserved Thr54 and Asn89 in the putative membrane-binding surface are essential for lipid selectivity and membrane targeting both in vitro and in the cell. Also, phosphorylation of Ser225 enhances the membrane affinity and plasma membrane selectivity of hSK1, presumably by modulating the interaction of Thr54 and Asn89 with the membrane. Collectively, these studies suggest that the specific plasma membrane localization and activation of SK1 is mediated largely by specific lipid protein interactions. PMID- 16243847 TI - Metallothionein disulfides are present in metallothionein-overexpressing transgenic mouse heart and increase under conditions of oxidative stress. AB - Metallothionein (MT) releases zinc under oxidative stress conditions in cultured cells. The change in the MT molecule after zinc release in vivo is unknown although in vitro studies have identified MT disulfide bond formation. The present study was undertaken to test the hypothesis that MT disulfide bond formation occurs in vivo. A cardiac-specific MT-overexpressing transgenic mouse model was used. Mice were administered saline as a control or doxorubicin (20 mg/kg), which is an effective anticancer drug but with severe cardiac toxicity at least partially because of the generation of reactive oxygen species. A differential alkylation of cysteine residues in MT of the heart extracts was performed. Free and metal-bound cysteines were first trapped by N-ethylmaleimide and the disulfide bonds were reduced by dithiothreitol followed by alkylation with radiolabeled iodoacetamide. Analyses of the differentially alkylated MTs in the heart extract by high performance liquid chromatography, SDS-PAGE, Western blot, and mass spectrometry revealed that disulfide bonds were present in MT in vivo under both physiological and oxidative stress conditions. More disulfide bonds were found in MT under the oxidative stress conditions. The MT disulfide bonds were likely intramolecular and both alpha- and beta-domains were involved in the disulfide bond formation, although the alpha-domain appeared to be more easily oxidized than the beta-domain. The results suggest that under physiological conditions, the formation of MT disulfide bonds is involved in the regulation of zinc homeostasis. Additional zinc release from MT under oxidative stress conditions is accompanied by more MT disulfide bond formation. PMID- 16243848 TI - Age at retirement and long term survival of an industrial population: prospective cohort study. AB - OBJECTIVE: To assess whether early retirement is associated with better survival. DESIGN: Long term prospective cohort study. SETTING: Petroleum and petrochemical industry, United States. SUBJECTS: Past employees of Shell Oil who retired at ages 55, 60, and 65 between 1 January 1973 and 31 December 2003. MAIN OUTCOME MEASURE: Hazard ratio of death adjusted for sex, year of entry to study, and socioeconomic status. RESULTS: Subjects who retired early at 55 and who were still alive at 65 had a significantly higher mortality than those who retired at 65 (hazard ratio 1.37, 95% confidence interval 1.09 to 1.73). Mortality was also significantly higher for subjects in the first 10 years after retirement at 55 compared with those who continued working (1.89, 1.58 to 2.27). After adjustment, mortality was similar between those who retired at 60 and those who retired at 65 (1.06, 0.92 to 1.22). Mortality did not differ for the first five years after retirement at 60 compared with continuing work at 60 (1.04, 0.82 to 1.31). CONCLUSIONS: Retiring early at 55 or 60 was not associated with better survival than retiring at 65 in a cohort of past employees of the petrochemical industry. Mortality was higher in employees who retired at 55 than in those who continued working. PMID- 16243850 TI - Reduction of anisotropic image resolution in transmission electron microtomography by use of quadrangular prism-shaped section. AB - A quadrangular prism specimen (a 'prism-shaped' section) was observed in three dimension (3-D) by transmission electron microtomography (TEMT) over as wide a tilt range as possible. Two types of specimens were prepared for the TEMT experiments: (i) a prism-shaped section of a block copolymer nanostructure, whose cross section was 200 nm on each side and (ii) a conventional ultrathin section having the same thickness (approximately 200 nm) as the prism-shaped section. Image quality of the projections taken at high tilt angles, e.g. 60 degrees, of the prism-shaped section was considerably better than that of the ultrathin section. This was because the path length of electron beam of the prism-shaped section was shorter (and hence the transmission was higher) than that of the ultrathin section at the same tilt angle. Thus, although the projections of the ultrathin section at the tilt angle larger than approximately 40 degrees did not effectively contribute to the 3-D reconstruction, those of the prism-shaped section at very high tilt angles, e.g. +/-75 degrees, can still be used for the 3 D reconstruction. Three orthogonal cross-sectional views of the 3-D reconstruction were extensively compared between the two sections. It was found that (i) the image contrast of the 3-D reconstruction was significantly enhanced and (ii) an elongation of structural object due to limitation of the angular range in the electron tomography was considerably reduced using the prism-shaped section. PMID- 16243849 TI - Prevention of colorectal cancer by colonoscopic surveillance in individuals with a family history of colorectal cancer: 16 year, prospective, follow-up study. AB - OBJECTIVE: To determine to what extent individuals with various family histories of colorectal cancer (from one to three or more affected first degree relatives) benefit from colonoscopic surveillance. DESIGN: Prospective, observational study of high risk families, followed up over 16 years. SETTING: Tertiary referral family cancer clinic in London. PARTICIPANTS: 1678 individuals from families registered with the clinic. Individuals were classified according to the strength of their family history: hereditary non-polyposis colorectal cancer (if they fulfilled the Amsterdam criteria), and one, two, or three affected first degree relatives (moderate risk). INTERVENTIONS: Colonoscopy was initially offered at five year intervals or three year intervals if an adenoma was detected. MAIN OUTCOME MEASURES: The incidence of adenomas with high risk pathological features or cancer. This was analysed by age, the extent of the family history, and findings on previous colonoscopies. The cohort was flagged for cancer and death. Incidence of colorectal cancer and mortality during over 15,000 person years of follow-up were compared with those expected in the absence of surveillance. RESULTS: High risk adenomas and cancer were most common in families with hereditary non-polyposis colorectal cancer (on initial colonoscopy 5.7% and 0.9%, respectively). In the families with moderate risk, these findings were particularly uncommon under age 45 (1.1% and 0%) and on follow-up colonoscopy if advanced neoplasia was absent initially (1.7% and 0.1%). The incidence of colorectal cancer was substantially lower-80% in families with moderate risk (P = 0.00004), and 43% in families with hereditary non-polyposis colorectal cancer (P = 0.06)-than the expected incidence in the absence of surveillance when the family history was taken into account. CONCLUSIONS: Colonoscopic surveillance reduces the risk of colorectal cancer in people with a strong family history. This study confirms that members of families with hereditary non-polyposis colorectal cancer require surveillance with short intervals. Individuals with a lesser family history may not require surveillance under age 45, and if advanced neoplasia is absent on initial colonoscopy, surveillance intervals may be lengthened. This would reduce the demand for colonoscopic surveillance. PMID- 16243851 TI - NADP-malate dehydrogenase gene evolution in Andropogoneae (Poaceae): gene duplication followed by sub-functionalization. AB - BACKGROUND AND AIMS: Plastid NADP-dependent malate dehydrogenase (MDH) catalyses the conversion of oxaloacetate to malate. In C4 plants, it is involved in photosynthetic carbon assimilation. In Poaceae, one NADP-MDH gene has been identified in rice (C3; Erhartoideae) and maize (C4; Panicoideae), whereas two tandemly repeated genes have been identified in Sorghum (C4; Panicoideae). In the present study, the molecular evolution of the NADP-MDH multigene family was investigated in order to analyse how the C4 isoform has evolved over a broader range of panicoid grasses. METHODS: Polymerase chain reaction (PCR)-based cloning was used to isolate cDNAs encoding NADP-MDHs from 15 species of Panicoideae. A gene phylogeny was reconstructed based on cDNA sequences using distance and maximum parsimony methods. Episodic selection along some branches of the phylogenetic tree was tested by analysing non-synonymous and synonymous rate ratios. Transcription of NADP-MDH genes was compared in green leaves of five accessions of Saccharum, Sorghum and Vetiveria using a semi-quantitative PCR approach. KEY RESULTS: Phylogenetic analyses of these data support the existence of two NADP-MDH gene lineages (NMDH-I and NMDH-II) in several Andropogoneae (i.e. Saccharum, Sorghum and Vetiveria). Episodic positive selection was shown along the basal branch of the NMDH-II clade. Three amino acid modifications allow the two gene lineages to be distinguished, suggesting a positive selection at these sites. In green leaves, we showed that the transcript accumulation was higher for NMDH-I than for NMDH-II. CONCLUSIONS: It is hypothesized that the maintenance of both NADP-MDH genes in some Andropogoneae is due to a partition of the original functions across both copies. NMDH-I probably corresponds to the C4 isoform as previously suggested. Nevertheless, some C4 species (e.g. maize) only have one gene which should be selected for its high expression level in leaves. This study confirms that gene duplicates have been recruited for C4 photosynthesis but are not required in every case. PMID- 16243852 TI - Nuclear DNA variation, chromosome numbers and polyploidy in the endemic and indigenous grass flora of New Zealand. AB - BACKGROUND AND AIMS: Little information is available on DNA C-values for the New Zealand flora. Nearly 85 % of the named species of the native vascular flora are endemic, including 157 species of Poaceae, the second most species-rich plant family in New Zealand. Few C-values have been published for New Zealand native grasses, and chromosome numbers have previously been reported for fewer than half of the species. The aim of this research was to determine C-values and chromosome numbers for most of the endemic and indigenous Poaceae from New Zealand. SCOPE: To analyse DNA C-values from 155 species and chromosome numbers from 55 species of the endemic and indigenous grass flora of New Zealand. KEY RESULTS: The new C values increase significantly the number of such measurements for Poaceae worldwide. New chromosome numbers were determined from 55 species. Variation in C value and percentage polyploidy were analysed in relation to plant distribution. No clear relationship could be demonstrated between these variables. CONCLUSIONS: A wide range of C-values was found in the New Zealand endemic and indigenous grasses. This variation can be related to the phylogenetic position of the genera, plants in the BOP (Bambusoideae, Oryzoideae, Pooideae) clade in general having higher C-values than those in the PACC (Panicoideae, Arundinoideae, Chloridoideae + Centothecoideae) clade. Within genera, polyploids typically have smaller genome sizes (C-value divided by ploidy level) than diploids and there is commonly a progressive decrease with increasing ploidy level. The high frequency of polyploidy in the New Zealand grasses was confirmed by our additional counts, with only approximately 10 % being diploid. No clear relationship between C value, polyploidy and rarity was evident. PMID- 16243853 TI - Interleukin-11 inhibits expression of insulin-like growth factor binding protein 5 mRNA in decidualizing human endometrial stromal cells. AB - Differentiation of endometrial stromal cells into decidual cells is essential for successful embryo implantation. Interleukin (IL)-11 signalling is critical for normal decidualization in the mouse. The expression of IL-11 and its receptors during the menstrual cycle, and the effect of exogenous IL-11 on the decidualization of human endometrial stromal cells in vitro, suggests a role for this cytokine in human decidualization. As the downstream target genes of IL-11 are also likely to be critical mediators of this process, this study aimed to identify genes regulated by IL-11 in decidualizing human endometrial stromal cells in vitro. Stromal cells isolated from endometrial biopsies were decidualized with 17beta estradiol (E) and medroxyprogesterone acetate (EP) in the presence or absence of exogenous IL-11, and total RNA used for cDNA microarray analysis and real-time RT-PCR. Microarray analysis revealed 16 up regulated and 11 down-regulated cDNAs in EP + IL-11-treated compared with EP treated cells. The most down-regulated gene was insulin-like growth factor binding protein-5 (IGFBP-5) (3.6-fold). Using real-time RT-PCR, IL-11 was confirmed to decrease IGFBP-5 transcript abundance 102-fold (P = 0.016; n = 6). No difference in IGFBP-5 immunostaining intensity was detected in stromal cells decidualized in the presence or absence of IL-11, and there was no effect of exogenous IGFBP-5 on the progression of steroid-induced in vitro decidualization. Interactions between IL-11 and its target genes, including IGFBP-5, may contribute to the regulation of decidualization and/or mediate communication between the decidua and invading trophoblast at implantation. PMID- 16243854 TI - Markedly elevated neonatal immunoreactive trypsinogen levels in the absence of cystic fibrosis gene mutations is not an indication for further testing. AB - AIMS: To investigate the immunoreactive trypsinogen (IRT) values above the usual 99th centile laboratory cut-off and determine the value of offering further testing to those infants with a markedly elevated IRT but no cystic fibrosis transmembrane regulator (CFTR) gene mutation identified by the screening programme. METHODS: All babies born in Victoria, Australia, between 1991 and 2003, were screened by IRT followed by CF gene mutation analysis. RESULTS: Of the 806,520 babies born, 9268 with the highest IRT levels had CFTR mutation analysis. There were 123 DeltaF508 homozygotes and 703 heterozygotes (86 with CF, 617 carriers). A total of 8442 babies had no CFTR gene mutation, of whom 18 (0.21%) had CF. The total number of CF babies with IRT greater than the laboratory cut off was 227 (2.4%). The IRT results of the CF patients were distributed normally, with the majority above the laboratory cut-off of newborn IRT results. There was no evidence of an excess of babies with CF in the very highest levels of IRT above the 99th centile. CONCLUSIONS: Only a small proportion of babies with a neonatal IRT >99th centile have CF. Additional CF testing for infants with an elevated IRT but no CFTR gene mutation has an extremely low yield, no matter how high the IRT result. PMID- 16243855 TI - An 8 year study of risk factors for SIDS: bed-sharing versus non-bed-sharing. AB - BACKGROUND: It is unclear if it is safe for babies to bed share with adults. In Ireland 49% of sudden infant death syndrome (SIDS) cases occur when the infant is bed-sharing with an adult. OBJECTIVE: To evaluate the effect of bed-sharing during the last sleep period on risk factors for SIDS in Irish infants. DESIGN: An 8 year (1994-2001) population based case control study of 287 SIDS cases and 831 controls matched for date, place of birth, and sleep period. Odds ratios and 95% confidence intervals were calculated by conditional logistic regression. RESULTS: The risk associated with bed-sharing was three times greater for infants with low birth weight for gestation (UOR 16.28 v 4.90) and increased fourfold if the combined tog value of clothing and bedding was > or =10 (UOR 9.68 v 2.34). The unadjusted odds ratio for bed-sharing was 13.87 (95% CI 9.58 to 20.09) for infants whose mothers smoked and 2.09 (95% CI 0.98 to 4.39) for non-smokers. Age of death for bed-sharing and sofa-sharing infants (12.8 and 8.3 weeks, respectively) was less than for infants not sharing a sleep surface (21.0 weeks, p<0.001) and fewer bed-sharing cases were found prone (5% v 32%; p = 0.001). CONCLUSION: Risk factors for SIDS vary according to the infant's sleeping environment. The increased risk associated with maternal smoking, high tog value of clothing and bedding, and low z scores of weight for gestation at birth is augmented further by bed-sharing. These factors should be taken into account when considering sleeping arrangements for young infants. PMID- 16243856 TI - Differing priorities for medical research funding. PMID- 16243857 TI - Paediatric cardiac resuscitation: can we do better? PMID- 16243859 TI - Is protecting children bad for your health? PMID- 16243858 TI - Diagnosing tuberculosis. PMID- 16243860 TI - Kayser Fischer ring. PMID- 16243861 TI - Patterns of research activity related to government policy: a UK web based survey. AB - AIMS: To describe the patterns of child and family health and wellbeing research activity in the fiscal year (FY) 2002/2003 in relation to UK government policies. METHODS: Projects investigating the health and wellbeing of children and families were located through a web based survey of major research funders, including UK government departments and non-departmental public bodies, research councils, and medical charities. A budget was estimated for each project for the FY 2002/2003, and each project coded according to a framework which reflected government priorities and research methodologies. RESULTS: There was a substantial amount of project information posted on the websites of the funding organisations, but the level of detail varied. For the FY 2002/2003, 31 organisations were identified that commissioned 567 projects investigating the health and wellbeing of children and families. Based on information from organisations' websites, this represented approximately 3% of their research budgets. Within this funding area, low proportions of research activity related to health inequalities (9% of total expenditure on child and family health research), health economic analysis (8%), primary and secondary prevention (12%), and children and adolescents at high risk of ill health (14%). CONCLUSIONS: A limited amount of research activity on children and families health funded in the FY 2002/2003 is addressing UK government policy priorities. This suggests the need to commission further research to fill gaps in the evidence. PMID- 16243862 TI - Burnout, psychological morbidity, job satisfaction, and stress: a survey of Canadian hospital based child protection professionals. AB - AIMS: (1) To measure the prevalence of burnout, psychological morbidity, job satisfaction, job stress, and consideration of alternate work among multidisciplinary hospital based child and youth protection (CYP) professionals; (2) to understand the relations between these variables; and (3) to understand the reasons for leaving among former programme members. METHODS: Mailed survey of current and former members of all Canadian academic hospital based CYP programmes. Surveys for current members contained validated measures of burnout, psychological morbidity, job satisfaction/stress, and questions about consideration of alternate work. Surveys for former members examined motivation(s) for leaving. RESULTS: One hundred and twenty six of 165 current members (76.4%) and 13/14 (92.9%) former members responded. Over one third (34.1%) of respondents exhibited burnout while psychological morbidity was present in 13.5%. Job satisfaction was high, with 68.8% finding their job "extremely" or "quite" satisfying, whereas 26.2% found their job "extremely" or "quite" stressful. Psychological morbidity, job satisfaction, and job stress were not associated with any of the demographic variables measured, but burnout was most prevalent among non-physician programme members. Almost two thirds of current members indicated that they had seriously considered a change in work situation. Former members indicated that burnout and high levels of job stress were most responsible for their decision to leave and that increasing the number of programme staff and, consequently, reducing the number of hours worked would have influenced their decision to stay. CONCLUSIONS: Current levels of burnout and the large proportion of individuals who have contemplated leaving the service suggest a potential crisis in Canadian hospital based CYP services. PMID- 16243863 TI - Klippel-Trenaunay syndrome. PMID- 16243864 TI - Developmental delay: when to suspect and how to investigate for an inborn error of metabolism. AB - The purpose of this review is to provide a practical guideline on the suspicion and investigation of inborn errors of metabolism (IEMs) as cause of developmental delay. Developmental delay is a common paediatric problem. Inborn errors of metabolism are a rare cause of developmental delay. However, it is important to detect IEMs for several reasons: accurate counselling may be given regarding recurrence risk; metabolic decompensation may be avoided; and specific treatments may be available. Certain clinical situations are more likely to point to an IEM as the cause of developmental delay. This review highlights the risk factors in the history, the important examination findings, and the appropriate biochemical investigation of the child with developmental delay. Following these guidelines makes "missing" an IEM unlikely. PMID- 16243865 TI - Erythema induratum of Bazin and episcleritis in a 6 year old girl. PMID- 16243866 TI - Adolescent medicine in paediatric practice. AB - Adolescents are a growing area in paediatric practice in both hospital and community settings. They make up around one quarter of the practice of many paediatricians. Yet until recently there has been little formal interest in young people's health in the UK. The situation is now changing, particularly following the publication of the "National Service Framework for children, young people and maternity services", which places a major emphasis on adolescent health. Given that this area is relatively new to many paediatricians, this article aims to provide an overview of the range of health problems that affect young people, to provide practical advice for working with this group in paediatric practice, and to outline current and future opportunities for training in adolescent health in the UK. PMID- 16243867 TI - Assessment of hypoxia in children with cystic fibrosis. AB - Hypoxia during sleep and exercise may occur in an important number of patients with cystic fibrosis (CF). Despite its recognition, no clear definition for hypoxia in CF exists, and nor do guidelines for commencing oxygen therapy. CF patients with hypoxia may have increased pulmonary artery pressure, reduced exercise ability, and skeletal muscle strength, and most importantly of all worse sleep quality, and a worse quality of life. Laboratory and rodent evidence exists to suggest that hypoxia may contribute to the decline in lung function in CF by upregulating lung inflammation, and encouraging growth of Pseudomonas aeruginosa, the most important pathogen associated with CF lung disease. The effects of hypoxia in childhood CF need to be fully studied, and a potential expanded role for oxygen as therapy in CF may be worthy of exploration. PMID- 16243868 TI - The impact of hormone physiology on clinical practice with reference to two papers spanning 40 years of endocrinology. AB - Following the pioneering work of Roger Ekins in describing the methodology for radioimmunoassay of hormone concentrations in plasma a series of publications followed describing variation in plasma hormone concentrations in a number of diseases. Though fluctuations in hormonal concentrations had been suspected, it was not until Hunter and Rigal documented these for the GH axis that it became apparent that the preferred mode of signalling for many hormone systems and in particular the anterior-pituitary hormones was of a pulsatile nature. PMID- 16243869 TI - Reduction of paediatric in-patient cardiac arrest and death with a medical emergency team: preliminary results. AB - AIMS: To determine the impact of a paediatric medical emergency team (MET) on cardiac arrest, mortality, and unplanned admission to intensive care in a paediatric tertiary care hospital. METHODS: Comparison of the retrospective incidence of cardiac arrest and death during 41 months before introduction of a MET service with the prospective incidence of these events during 12 months after its introduction. Comparison of transgression of MET call criteria in patients who arrested and died before and after introduction of MET. RESULTS: Cardiac arrest decreased from 20 among 104 780 admissions (0.19/1000) to 4 among 35 892 admissions (0.11/1000) (risk ratio 1.71, 95% CI 0.59 to 5.01), while death decreased from 13 (0.12/1000) to 2 (0.06/1000) during these periods (risk ratio 2.22, 95% CI 0.50 to 9.87). Unplanned admissions to intensive care increased from 20 (SD 6) to 24 (SD 9) per month. The incidence of transgression of MET call criteria in patients who arrested decreased from 17 to 0 (risk difference 0.16/1000, 95% CI 0.09 to 0.24), and in those who died, decreased from 12 to 0 (risk difference 0.11/1000, 95% CI 0.05 to 0.18) after introduction of MET. CONCLUSIONS: Introduction of a medical emergency team service was coincident with a reduction of cardiac arrest and mortality and a slight increase in admissions to intensive care. PMID- 16243870 TI - Diagnostic accuracy of chest radiography in detecting mediastinal lymphadenopathy in suspected pulmonary tuberculosis. AB - OBJECTIVE: To estimate the diagnostic accuracy of chest radiography in the detection of chest lymphadenopathy in children with clinically suspected pulmonary tuberculosis. METHODS: DESIGN: Prospective cross sectional study. SETTING: A short stay ward in a children's hospital in South Africa. PATIENTS: Consecutive children under 14 years of age admitted with suspected pulmonary tuberculosis. DIAGNOSTIC TEST: Antero-posterior and/or lateral chest x rays interpreted independently and blind to the reference standard by three primary care clinicians and three paediatricians, all with a special interest in tuberculosis. Reference standard: Spiral chest computed tomography (CT) with contrast injection. RESULTS: One hundred children (median age 21.5 months) were enrolled. Lymphadenopathy was present in 46 of 100 reference CT scans and judged to be present in 47.1% of x ray assessments. Overall sensitivity was 67% and specificity 59%. Primary care clinicians were more sensitive (71.5% v 63.3%, p = 0.047) and less specific (49.8% v 68.9%, p<0.001) than paediatricians. Overall accuracy was higher for the paediatricians (diagnostic odds ratio 3.83 v 2.49, p = 0.008). The addition of a lateral to an antero-posterior view did not significantly increase accuracy (diagnostic odds ratio 3.09 v 3.73, p = 0.16). Chance adjusted inter-observer agreement (kappa) varied widely between viewer pairs, but was around 30%. CONCLUSIONS: Detection of mediastinal lymphadenopathy on chest x ray to diagnose pulmonary tuberculosis in children must be interpreted with caution. Diagnostic accuracy might be improved by refining radiological criteria for lymphadenopathy. PMID- 16243871 TI - Tuberculosis in New Zealand, 1992-2001: a resurgence. AB - OBJECTIVE: To describe the recent epidemiology and clinical features of paediatric tuberculosis (TB) in New Zealand (NZ). METHODS: A retrospective review was conducted of clinical, laboratory, and radiology records of children <16 years old diagnosed with TB between January 1992 and June 2001 in nine NZ health districts. RESULTS: A total of 274 patients <16 years old were identified; the average annual TB rate was 4.8 per 100,000. Rates rose over time reaching a peak of 10.1 in 1999. Rates were highest in under-5 year olds, at 6.2 per 100,000, and varied by ethnicity: African 575.2, Pacific Island 15.2, Maori 6.4, Asian 5.6, and European 0.6. Seventy two cases (26%) were foreign born. Thirty six per cent of cases were not detected until they presented with symptoms and of these 44% had no known TB contact. Most cases were identified by contact tracing (48%) or immigrant screening (11%); 43% were part of outbreaks. Miliary TB or meningitis occurred in 8% of patients, two of whom died. Drug resistance was found in 7% of culture positive cases and no HIV co-infection was found. CONCLUSIONS: A resurgence of TB occurred among children in NZ between 1992 and 2001 predominantly involving non-European and immigrant groups. Despite established contact tracing and immigrant screening programmes, many cases were part of outbreaks, remained unidentified until symptoms arose, or had no known TB contact. These findings point to an unrecognised burden of adult disease, ongoing community transmission, and missed opportunities for prevention. Further study is required to confirm these hypotheses. PMID- 16243873 TI - The significance of elevated CSF lactate. AB - The final diagnosis of 158 patients who had a cerebrospinal fluid (CSF) lactate concentration greater than 2 mmol/l was ascertained. The conditions included seizures, inflammatory changes, and proven metabolic disorders. For the diagnosis of congenital lactic acidoses, CSF lactate should ideally be measured in a seizure free patient after any acute illness. PMID- 16243872 TI - The prevalence of symptoms associated with pulmonary tuberculosis in randomly selected children from a high burden community. AB - BACKGROUND: Diagnosis of childhood tuberculosis is problematic and symptom based diagnostic approaches are often promoted in high burden settings. This study aimed (i) to document the prevalence of symptoms associated with tuberculosis among randomly selected children living in a high burden community, and (ii) to compare the prevalence of these symptoms in children without tuberculosis to those in children with newly diagnosed tuberculosis. METHODS: A cross sectional, community based survey was performed on a 15% random sample of residential addresses. A symptom based questionnaire and tuberculin skin test (TST) were completed in all children. Chest radiographs were performed according to South African National Tuberculosis Control Program guidelines. RESULTS: Results were available in 1415 children of whom 451 (31.9%) were TST positive. Tuberculosis was diagnosed in 18 (1.3%) children. Of the 1397 children without tuberculosis, 253 (26.4%) reported a cough during the preceding 3 months. Comparison of individual symptoms (cough, dyspnoea, chest pain, haemoptysis, anorexia, weight loss, fatigue, fever, night sweats) in children with and without tuberculosis revealed that only weight loss differed significantly (OR = 4.5, 95% CI 1.5 to 12.3), while the combination of cough and weight loss was most significant (OR = 5.4, 95% CI 1.7 to 16.9). Children with newly diagnosed tuberculosis reported no symptoms in 50% of cases. CONCLUSION: Children from this high burden community frequently reported symptoms associated with tuberculosis. These symptoms had limited value to differentiate children diagnosed with tuberculosis from those without tuberculosis. Improved case definitions and symptom characterisation are required when evaluating the diagnostic value of symptoms. PMID- 16243874 TI - Muscle fibre type grouping in high resolution ultrasound. PMID- 16243876 TI - A memorable stay. PMID- 16243875 TI - Guidelines and adenosine dosing in supraventricular tachycardia. AB - This retrospective review of present practice of administration of adenosine by paediatricians shows that current guidelines recommend starting doses that are effective in only 9% of infants and children. PMID- 16243877 TI - Postural hallucinations? An unusual presentation of anaemia. AB - The case of a 14 year old girl is reported, referred to Child Psychiatry with tearfulness and possible auditory hallucinations. Closer questioning revealed a history of low energy, sleeping excessively without refreshment, exertional dyspnoea, and poor growth. Psychologically, there was no evidence of low mood or negative cognitions despite the inexplicable tearfulness. Detailed enquiry revealed the "hallucinations" to be "whooshing" noises in her ears precipitated by standing. She was found to have a haemoglobin level of 55 g/l, attributed to a combination of poor diet and menorrhagia.Periodic, anaemic, cerebral hypoxia could be proposed to be the root of most, if not all, of the symptoms, illustrating the importance of marrying physical and psychological history taking with suitable investigations for an eminently treatable condition. PMID- 16243879 TI - Randomisation. PMID- 16243878 TI - Superglue mistaken for eye drops. PMID- 16243880 TI - Towards evidence based medicine for paediatricians. PMID- 16243881 TI - Should steroids be used in children with meningococcal shock? PMID- 16243882 TI - Should children with Henoch-Schonlein purpura and abdominal pain be treated with steroids? PMID- 16243883 TI - Do cuffed endotracheal tubes increase the risk of airway mucosal injury and post extubation stridor in children? PMID- 16243884 TI - Biopsychosocial approach to functional abdominal pain. PMID- 16243885 TI - Clinical assessment of neonatal hyperbilirubinaemia. PMID- 16243887 TI - Melatonin and epilepsy. PMID- 16243886 TI - Nutritional rickets is increasingly diagnosed in children of ethnic origin. PMID- 16243888 TI - Is timing of haemorrhage by spectrophotometry similar for haemorrhages in the subdural and subarachnoid space? PMID- 16243889 TI - Community needlestick injuries may still be dangerous. PMID- 16243890 TI - More evidence is needed in the antibiotic treatment of Pseudomonas aeruginosa colonisation. PMID- 16243891 TI - A vaccine scare in 19th century Northampton. PMID- 16243892 TI - Car seat safety for premature and LBW infants. PMID- 16243894 TI - Melatonin: prescribing practices and adverse events. PMID- 16243893 TI - Interpreting immunogenicity data in UK studies. PMID- 16243895 TI - Food challenge tests. PMID- 16243896 TI - Hearing impairment: age at diagnosis, severity, and language outcomes. PMID- 16243897 TI - Cytotoxicity of human RNase-based immunotoxins requires cytosolic access and resistance to ribonuclease inhibition. AB - Immunotoxins are targeted therapeutics designed to kill cancer cells. The targeting moiety of an immunotoxin selectively binds to a tumor cell and targets it for death via an attached toxin. Because the toxins are typically of plant or bacterial origin, their clinical use is limited by immunogenicity and nonspecific toxicity. To circumvent these problems, we have begun to engineer immunotoxins containing human pancreatic ribonuclease. Here we describe the generation of ribonuclease mutants designed to evade a ubiquitous cytosolic inhibitor that would otherwise block cytotoxicity. Two mutants retained catalytic activity and were relatively resistant to the inhibitor. To deliver them to human T leukemic cells, these ribonuclease variants were fused to a single chain Fv fragment specific for CD 7. The ribonuclease-sFv fusion proteins bound CD 7(+) T cells and were internalized yet were not cytotoxic. Transfection of the proteins directly into the cytosol reduced cell viability, suggesting that the failure of the immunotoxins to kill cells when added externally resulted from the inability of the ribonuclease moiety to access the cytosol efficiently. Our results indicate appropriate intracellular routing, as well as resistance to inhibition, is critical to the cytotoxicity of human ribonuclease-based immunotoxins. PMID- 16243898 TI - A set of multicolored Photinus pyralis luciferase mutants for in vivo bioluminescence applications. AB - Error-prone PCR was used to isolate Photinus pyralis luciferase mutants producing bright light in the red-orange region of the spectrum. All mutations were clustered in the beta5-alpha10-beta6 region of N-terminal subdomain B and appear to affect bioluminescence color by modulating the position of the Ser314-Leu319 mobile loop with respect to the putative active site. Two red variants (Q283R and S284G) and one orange mutant (S293P) contained a single substitution. Although the remaining orange variant contained two mutations, L287I mainly contributed to the color change. Emission spectra collected on whole cells at pH 7.0 revealed that while a single peak of lambdamax approximately 605 nm accounts for red light production by the Q283R and S284G variants, orange light results from the contribution of two peaks of lambdamax approximately 560 and 600 nm. All spectra underwent a red-shift when cells were assayed under acidic conditions, whereas a blue-shift was observed at pH 8.0, indicating that the internal pH of Escherichia coli is close to the external pH shortly after imposition of acid or alkaline stress. In addition, changes in assay pH led to bimodal emission spectra, lending support to the idea that bioluminescence color is determined by the relative contribution of yellow-green and red-orange peaks. The set of multicolored luciferase mutants described here may prove useful for a variety of applications including biosensing, pH monitoring, and tissue and animal imaging. PMID- 16243899 TI - Thromboxane A2 induces airway constriction through an M3 muscarinic acetylcholine receptor-dependent mechanism. AB - Thromboxane A2 (TXA2) is a potent lipid mediator released by platelets and inflammatory cells and is capable of inducing vasoconstriction and bronchoconstriction. In the airways, it has been postulated that TXA2 causes airway constriction by direct activation of thromboxane prostanoid (TP) receptors on airway smooth muscle cells. Here we demonstrate that although TXA2 can mediate a dramatic increase in airway smooth muscle constriction and lung resistance, this response is largely dependent on vagal innervation of the airways and is highly sensitive to muscarinic acetylcholine receptor (mAChR) antagonists. Further analyses employing pharmacological and genetic strategies demonstrate that TP-dependent changes in lung resistance and airway smooth muscle tension require expression of the M2 mAChR subtype. These results raise the possibility that some of the beneficial actions of anticholinergic agents used in the treatment of asthma and chronic obstructive pulmonary disease result from limiting physiological changes mediated through the TP receptor. Furthermore, these findings demonstrate a unique pathway for TP regulation of homeostatic mechanisms in the airway and suggest a paradigm for the role of TXA2 in other organ systems. PMID- 16243900 TI - Impaired NO-dependent inhibition of store- and receptor-operated calcium entry in pulmonary vascular smooth muscle after chronic hypoxia. AB - We have recently demonstrated that chronic hypoxia (CH) attenuates nitric oxide (NO)-mediated decreases in pulmonary vascular smooth muscle (VSM) intracellular free calcium concentration ([Ca2+]i) and promotes NO-dependent VSM Ca2+ desensitization. The objective of the current study was to identify potential mechanisms by which CH interferes with regulation of [Ca2+]i by NO. We hypothesized that CH impairs NO-mediated inhibition of store-operated (capacitative) Ca2+ entry (SOCE) or receptor-operated Ca2+ entry (ROCE) in pulmonary VSM. To test this hypothesis, we examined effects of the NO donor, spermine NONOate, on SOCE resulting from depletion of intracellular Ca2+ stores with cyclopiazonic acid, and on UTP-induced ROCE in isolated, endothelium denuded, pressurized pulmonary arteries (213 +/- 8 microm inner diameter) from control and CH (4 wk at 0.5 atm) rats. Arteries were loaded with fura-2 AM to continuously monitor VSM [Ca2+]i. We found that the change in [Ca2+]i associated with SOCE and ROCE was significantly reduced in vessels from CH animals. Furthermore, spermine NONOate diminished SOCE and ROCE in vessels from control, but not CH animals. We conclude that NO-mediated inhibition of SOCE and ROCE is impaired after CH-induced pulmonary hypertension. PMID- 16243901 TI - Influence of pulmonary arterial endothelial cells on quinone redox status: effect of hyperoxia-induced NAD(P)H:quinone oxidoreductase 1. AB - The objective of this study was to examine the impact of chronic hyperoxic exposure (95% O2 for 48 h) on intact bovine pulmonary arterial endothelial cell redox metabolism of 2,3,5,6-tetramethyl-1,4-benzoquinone (duroquinone, DQ). DQ or durohydroquinone (DQH2) was added to normoxic or hyperoxia-exposed cells in air saturated medium, and the medium DQ concentrations were measured over 30 min. DQ disappeared from the medium when DQ was added and appeared in the medium when DQH2 was added, such that after approximately 15 min, a steady-state DQ concentration was approached that was approximately 4.5 times lower for the hyperoxia-exposed than the normoxic cells. The rate of DQ-mediated reduction of the cell membrane-impermeant redox indicator, potassium ferricyanide [Fe(CN)6(3 )], was also approximately twofold faster for the hyperoxia-exposed cells. Inhibitor studies and mathematical modeling suggested that in both normoxic and hyperoxia-exposed cells, NAD(P)H:quinone oxidoreductase 1 (NQO1) was the dominant DQ reductase and mitochondrial electron transport complex III the dominant DQH2 oxidase involved and that the difference between the net effects of the cells on DQ redox status could be attributed primarily to a twofold increase in the maximum NQO1-mediated DQ reduction rate in the hyperoxia-exposed cells. Accordingly, NQO1 protein and total activity were higher in hyperoxia-exposed than normoxic cell cytosolic fractions. One outcome for hyperoxia-exposed cells was enhanced protection from cell-mediated DQ redox cycling. This study demonstrates that exposure to chronic hyperoxia increases the capacity of pulmonary arterial endothelial cells to reduce DQ to DQH2 via a hyperoxia-induced increase in NQO1 protein and total activity. PMID- 16243902 TI - Central apnoea after balanced general anaesthesia that included dexmedetomidine. AB - Dexmedetomidine is an alpha(2)-adrenoreceptor agonist that, in spite of its potent sedative, amnesic, and analgesic properties, has minimal respiratory depressant effect. Even at doses adequate for general anaesthesia, it does not cause central apnoea. Thus, it has been claimed that "combining alpha(2)-agonists with opiate narcotics or non-steroidal anti-inflammatory drugs can enhance the analgesic efficacy without increasing the respiratory depressant effect of the latter" and "the combination of alpha(2)-adrenoceptor agonists with opioids does not lead to further ventilatory depression". We present a case of central apnoea after general anaesthesia that included opioids and dexmedetomidine, and remind the readers that in susceptible patients, dexmedetomidine may cause life threatening respiratory depression through potentiation of co-administered central nervous system depressants. PMID- 16243903 TI - Transcriptome and selected metabolite analyses reveal multiple points of ethylene control during tomato fruit development. AB - Transcriptome profiling via cDNA microarray analysis identified 869 genes that are differentially expressed in developing tomato (Solanum lycopersicum) pericarp. Parallel phenotypic and targeted metabolite comparisons were employed to inform the expression analysis. Transcript accumulation in tomato fruit was observed to be extensively coordinated and often completely dependent on ethylene. Mutation of an ethylene receptor (Never-ripe [Nr]), which reduces ethylene sensitivity and inhibits ripening, alters the expression of 37% of these 869 genes. Nr also influences fruit morphology, seed number, ascorbate accumulation, carotenoid biosynthesis, ethylene evolution, and the expression of many genes during fruit maturation, indicating that ethylene governs multiple aspects of development both prior to and during fruit ripening in tomato. Of the 869 genes identified, 628 share homology (E-value < or = 1 x 10(-10)) with known gene products or known protein domains. Of these 628 loci, 72 share homology with previously described signal transduction or transcription factors, suggesting complex regulatory control. These results demonstrate multiple points of ethylene regulatory control during tomato fruit development and provide new insights into the molecular basis of ethylene-mediated ripening. PMID- 16243904 TI - PGP4, an ATP binding cassette P-glycoprotein, catalyzes auxin transport in Arabidopsis thaliana roots. AB - Members of the ABC (for ATP binding cassette) superfamily of integral membrane transporters function in cellular detoxification, cell-to-cell signaling, and channel regulation. More recently, members of the multidrug resistance P glycoprotein (MDR/PGP) subfamily of ABC transporters have been shown to function in the transport of the phytohormone auxin in both monocots and dicots. Here, we report that the Arabidopsis thaliana MDR/PGP PGP4 functions in the basipetal redirection of auxin from the root tip. Reporter gene studies showed that PGP4 was strongly expressed in root cap and epidermal cells. PGP4 exhibits apolar plasma membrane localization in the root cap and polar localization in tissues above. Root gravitropic bending and elongation as well as lateral root formation were reduced in pgp4 mutants compared with the wild type. pgp4 exhibited reduced basipetal auxin transport in roots and a small decrease in shoot-to-root transport consistent with a partial loss of the redirective auxin sink in the root. Seedlings overexpressing PGP4 exhibited increased shoot-to-root auxin transport. Heterologous expression of PGP4 in mammalian cells resulted in 1-N naphthylthalamic acid-reversible net uptake of [3H]indole-3-acetic acid. These results indicate that PGP4 functions primarily in the uptake of redirected or newly synthesized auxin in epidermal root cells. PMID- 16243905 TI - Functional differentiation of bundle sheath and mesophyll maize chloroplasts determined by comparative proteomics. AB - Chloroplasts of maize (Zea mays) leaves differentiate into specific bundle sheath (BS) and mesophyll (M) types to accommodate C4 photosynthesis. Consequences for other plastid functions are not well understood but are addressed here through a quantitative comparative proteome analysis of purified M and BS chloroplast stroma. Three independent techniques were used, including cleavable stable isotope coded affinity tags. Enzymes involved in lipid biosynthesis, nitrogen import, and tetrapyrrole and isoprenoid biosynthesis are preferentially located in the M chloroplasts. By contrast, enzymes involved in starch synthesis and sulfur import preferentially accumulate in BS chloroplasts. The different soluble antioxidative systems, in particular peroxiredoxins, accumulate at higher levels in M chloroplasts. We also observed differential accumulation of proteins involved in expression of plastid-encoded proteins (e.g., EF-Tu, EF-G, and mRNA binding proteins) and thylakoid formation (VIPP1), whereas others were equally distributed. Enzymes related to the C4 shuttle, the carboxylation and regeneration phase of the Calvin cycle, and several regulators (e.g., CP12) distributed as expected. However, enzymes involved in triose phosphate reduction and triose phosphate isomerase are primarily located in the M chloroplasts, indicating that the M-localized triose phosphate shuttle should be viewed as part of the BS-localized Calvin cycle, rather than a parallel pathway. PMID- 16243906 TI - Cell cycle progression in the pericycle is not sufficient for SOLITARY ROOT/IAA14 mediated lateral root initiation in Arabidopsis thaliana. AB - To study the mechanisms behind auxin-induced cell division, lateral root initiation was used as a model system. By means of microarray analysis, genome wide transcriptional changes were monitored during the early steps of lateral root initiation. Inclusion of the dominant auxin signaling mutant solitary root1 (slr1) identified genes involved in lateral root initiation that act downstream of the auxin/indole-3-acetic acid (AUX/IAA) signaling pathway. Interestingly, key components of the cell cycle machinery were strongly defective in slr1, suggesting a direct link between AUX/IAA signaling and core cell cycle regulation. However, induction of the cell cycle in the mutant background by overexpression of the D-type cyclin (CYCD3;1) was able to trigger complete rounds of cell division in the pericycle that did not result in lateral root formation. Therefore, lateral root initiation can only take place when cell cycle activation is accompanied by cell fate respecification of pericycle cells. The microarray data also yielded evidence for the existence of both negative and positive feedback mechanisms that regulate auxin homeostasis and signal transduction in the pericycle, thereby fine-tuning the process of lateral root initiation. PMID- 16243907 TI - Maize rough sheath2 and its Arabidopsis orthologue ASYMMETRIC LEAVES1 interact with HIRA, a predicted histone chaperone, to maintain knox gene silencing and determinacy during organogenesis. AB - Plant shoots are characterized by indeterminate growth resulting from the action of a population of stem cells in the shoot apical meristem (SAM). Indeterminacy within the SAM is specified in part by the class I knox homeobox genes. The myb domain proteins rough sheath2 (RS2) and ASYMMETRIC LEAVES1 (AS1) from maize (Zea mays) and Arabidopsis thaliana, respectively, are required to establish determinacy during leaf development. These proteins are part of a cellular memory system that in response to a stem cell-derived signal keeps knox genes in an off state during organogenesis. Here, we show that RS2/AS1 can form conserved protein complexes through interaction with the DNA binding factor ASYMMETRIC LEAVES2, a predicted RNA binding protein (RIK, for RS2-Interacting KH protein), and a homologue of the chromatin-remodeling protein HIRA. Partial loss of HIRA function in Arabidopsis results in developmental defects comparable to those of as1 and causes reactivation of knox genes in developing leaves, demonstrating a direct role for HIRA in knox gene repression and the establishment of determinacy during leaf formation. Our data suggest that RS2/AS1 and HIRA mediate the epigenetic silencing of knox genes, possibly by modulating chromatin structure. Components of this process are conserved in animals, suggesting the possibility that a similar epigenetic mechanism maintains determinacy during both plant and animal development. PMID- 16243909 TI - Exercise training increases basal tone in arterioles distal to chronic coronary occlusion. AB - Endurance exercise training increases basal active tone in coronary arteries and enhances myogenic tone in coronary arterioles of control animals. Paradoxically, exercise training has also been shown to augment nitric oxide production and nitric oxide-mediated relaxation in coronary arterioles. The purpose of the present study was to examine the effect of exercise training on basal active tone of arterioles (approximately 150 microm ID) isolated from the collateral dependent region of hearts exposed to chronic coronary occlusion. Ameroid occluders were surgically placed around the proximal left circumflex coronary artery of miniature swine. Arterioles were isolated from both the collateral dependent and nonoccluded myocardial regions of sedentary (pen confined) and exercise-trained (treadmill run; 14 wk) pigs. Coronary tone was studied in isolated arterioles using microvessel myographs and standard isometric techniques. Exposure to nominally Ca2+-free external solution reduced resting tension in all arterioles; decreases were most profound (P < 0.05) in arterioles from the collateral-dependent region of exercise-trained animals. Furthermore, nitric oxide synthase (NOS) inhibition (N(omega)-nitro-L-arginine methyl ester; 100 microM) unmasked markedly increased nitric oxide-sensitive tone in arterioles from the collateral-dependent region of exercise-trained swine. Blockade of K+ channels revealed significantly enhanced K+ channel contribution to basal tone in collateral-dependent arterioles of exercise-trained pigs. Protein content of endothelial NOS (eNOS) and phosphorylated eNOS (pS1179), determined by immunoblot, was elevated in arterioles from exercise-trained animals with the greatest effect in collateral-dependent vasculature. Taken together, we demonstrate the interaction of opposing exercise training-enhanced arteriolar basal active tone, nitric oxide production, and K+ channel activity in chronic coronary occlusion, potentially enhancing the capacity to regulate blood flow to collateral-dependent myocardium. PMID- 16243910 TI - Transcriptional analysis of doxorubicin-induced cardiotoxicity. AB - Doxorubicin is an effective chemotherapeutic agent against a broad range of tumors. However, a threshold dose of doxorubicin causes an unacceptably high incidence of heart failure and limits its clinical utility. We have established two models of doxorubicin cardiotoxicity in mice: 1) in an acute model, mice are treated with 15 mg/kg of doxorubicin once; and 2) in a chronic model, they receive 3 mg/kg weekly for 12 wk. Using echocardiography, we have monitored left ventricular function during treatment in the chronic model and seen the expected development of dilated cardiomyopathy. Treated mice showed histological abnormalities similar to those seen in patients with doxorubicin cardiomyopathy. To investigate transcriptional regulation in these models, we used a muscle specific cDNA microarray. We have identified genes that respond to doxorubicin exposure in both models and confirmed these results using real-time PCR. In the acute model, a set of genes is regulated early and rapidly returns to baseline levels, consistent with the half-life of doxorubicin. In the chronic model, which mimics the clinical situation much more closely, we identified dysregulated genes that implicate specific mechanisms of cardiac toxicity. These include STARS, a hypertrophy-responsive gene; SNF1-kinase, a potential modulator of ATP levels; and AXUD1, a downstream target of the proapoptotic regulator AXIN1. PMID- 16243908 TI - TRANSPARENT TESTA10 encodes a laccase-like enzyme involved in oxidative polymerization of flavonoids in Arabidopsis seed coat. AB - The Arabidopsis thaliana transparent testa10 (tt10) mutant exhibits a delay in developmentally determined browning of the seed coat, also called the testa. Seed coat browning is caused by the oxidation of flavonoids, particularly proanthocyanidins, which are polymers of flavan-3-ol subunits such as epicatechin and catechin. The tt10 mutant seeds accumulate more epicatechin monomers and more soluble proanthocyanidins than wild-type seeds. Moreover, intact testa cells of tt10 cannot trigger H2O2-independent browning in the presence of epicatechin and catechin, in contrast with wild-type cells. UV-visible light detection and mass spectrometry revealed that the major oxidation products obtained with epicatechin alone are yellow dimers called dehydrodiepicatechin A. These products differ from proanthocyanidins in the nature and position of their interflavan linkages. Flavonol composition was also affected in tt10 seeds, which exhibited a higher ratio of quercetin rhamnoside monomers versus dimers than wild-type seeds. We identified the TT10 gene by a candidate gene approach. TT10 encodes a protein with strong similarity to laccase-like polyphenol oxidases. It is expressed essentially in developing testa, where it colocalizes with the flavonoid end products proanthocyanidins and flavonols. Together, these data establish that TT10 is involved in the oxidative polymerization of flavonoids and functions as a laccase-type flavonoid oxidase. PMID- 16243911 TI - Calcium receptor is functionally expressed in rat neonatal ventricular cardiomyocytes. AB - Both intra- and extracellular calcium play multiple roles in the physiology and pathophysiology of cardiomyocytes, especially in stimulus-contraction coupling. The intracellular calcium level is closely controlled through the concerted actions of calcium channels, exchangers, and pumps; however, the expression and function(s) of the so-called calcium-sensing receptor (CaR) in the heart remain less well characterized. The CaR is a seven-transmembrane receptor, which, in response to noncovalent binding of extracellular calcium, activates intracellular effectors, including G proteins and extracellular signal-regulated kinases (ERK1/2). We have shown that cultured neonatal cardiomyocytes express the CaR messenger RNA and the CaR protein. Furthermore, increasing concentrations of extracellular calcium and a type II CaR activator "calcimimetic" caused inositol phosphate (IP) accumulation, downregulated tritiated thymidine incorporation, and supported ERK1/2 phosphorylation, suggesting that the CaR protein is functionally active. Interestingly, the calcimimetic induced a more rapid ERK1/2 phosphorylation than calcium and left-shifted the IP concentration-response curve for extracellular calcium, supporting the hypothesis that CaR is functionally expressed in cardiac myocytes. This notion was underscored by studies using a virus containing a dominant-negative CaR construct, because this protein blunted the calcium-induced IP response. In conclusion, we have shown that the CaR is functionally expressed in neonatal ventricular cardiomyocytes and that the receptor activates second messenger pathways, including IP and ERK, and decreases DNA synthesis. A specific calcium-sensing receptor on cardiac myocytes could play a role in regulating cardiac development, function, and homeostasis. PMID- 16243912 TI - Plasticity of GABAergic control of hypothalamic presympathetic neurons in hypertension. AB - Increased sympathetic outflow contributes to the pathogenesis of hypertension. However, the mechanisms of increased sympathetic drive in hypertension remain unclear. We examined the tonic GABAergic inhibition in control of the excitability of paraventricular (PVN) presympathetic neurons in spontaneously hypertensive rats (SHR) and normotensive controls, including Sprague-Dawley (SD) and Wistar-Kyoto (WKY) rats. Whole cell patch-clamp recordings were performed on retrogradely labeled PVN neurons projecting to the rostral ventrolateral medulla (RVLM) in brain slices. The basal firing rate of PVN neurons was significantly decreased in 13-wk-old SD and WKY rats but increased in 13-wk-old SHR, compared with their respective 6-wk-old controls. The GABA(A) antagonist bicuculline consistently increased the firing of PVN neurons in normotensive controls. Surprisingly, bicuculline either decreased the firing or had no effect in 59.3% of labeled cells in 13-wk-old SHR. In contrast, the GABA(B) antagonist CGP-55845 had no effect on the firing of PVN neurons in normotensive controls but significantly increased the firing of 75% of cells studied in 13-wk-old SHR. Furthermore, the evoked GABA(A) current decreased significantly in labeled PVN neurons of 13-wk-old SHR compared with that in normotensive controls. Both the frequency and amplitude of GABAergic spontaneously inhibitory postsynaptic currents were also reduced in 13-wk-old SHR. This study demonstrates an unexpected functional change in GABA(A) and GABA(B) receptors in regulation of the firing activity of PVN-RVLM neurons in SHR. This change in GABA(A) receptor function and GABAergic inputs to PVN output neurons may contribute to increased sympathetic outflow in hypertension. PMID- 16243913 TI - Respective contribution of age, mean arterial pressure, and body weight on central arterial distensibility in SHR. AB - In spontaneously hypertensive rats (SHR), carotid and aortic distensibilities measured at operational blood pressure (BP) are reduced. Increased body weight and mean arterial pressure (MAP) are both known to reduce distensibility independently. However, whether, after adjustment to body weight and mean BP, distensibility remains reduced in SHR has never been investigated. Carotid and abdominal aorta distensibilities were measured under anesthesia in SHR at 5, 12, 52, and 78 wk of age, and measurements were compared with age-matched normotensive Wistar rats. Each age group was composed of 9 or 10 animals. We determined distensibility using echo-tracking techniques of high resolution. Compared with Wistar rats, carotid and aortic distensibilities measured at operational MAP are reduced in SHR. This reduction is accentuated with age, particularly for the carotid artery. After adjustment to body weight and MAP, carotid and aortic distensibilities become identical in Wistar and SHR (or even slightly increased in SHR) but continue to be reduced with age, mainly for the carotid artery. In conclusion, in SHR, age and high BP do not have a parallel and similar influence on the reduction of arterial distensibility. Aging constantly reduces arterial distensibility, whereas MAP levels contribute to maintenance of arterial function. PMID- 16243914 TI - Sustained preconditioning induced by cardiac transgenesis with the tetracycline transactivator. AB - Preconditioning protocols that protect the heart from ischemic injury may aid in the development of new therapies. However, the temporal window of cardioprotection is limited to a few days after the preconditioning stimulus. Here we report a sustained cardioprotected phenotype in mice expressing a tetracycline transactivator (tTA) transcription factor under the control of the alpha-myosin heavy chain (alphaMHC) promoter. alphaMHC-tTA mice were originally designed for tetracycline-regulated gene expression in the heart (Tet system). However, we found that after 45 min of global ischemia at 37 degrees C, left ventricular developed pressure (LVDP) of Langendorff-perfused alphaMHC-tTA mouse hearts rapidly recovered in 5 min to 60% of initial levels, whereas LVDP of wild type (WT) littermates recovered to only 10% of the initial level. Improved postischemic recovery of function for alphaMHC-tTA hearts was associated with a 50% decrease of infarct size and a significantly smaller release of lactate dehydrogenase to the coronary effluent. Improved postischemic recovery was not attributable to differences in coronary flow that was similar for WT- and alphaMHC-tTA hearts during recovery. Moreover, improved postischemic recovery of alphaMHC-tTA hearts was not abolished by inhibitors of classical cardioprotective effectors (mitochondrial ATP-sensitive K+ channels, PKC, or adenosine receptors), suggesting a novel mechanism. Finally, the tetracycline analog doxycycline, which inhibits binding of tTA to DNA, did not abolish improved recovery for alphaMHC tTA hearts. The sustained cardioprotected phenotype of alphaMHC-tTA hearts may have implications for developing new therapies to minimize cardiac ischemic injury. Furthermore, investigations of cardioprotection using the Tet system may be aberrantly influenced by sustained preconditioning induced by cardiac transgenesis with tTA. PMID- 16243915 TI - Effect of thrombolysis on myocardial injury: recombinant tissue plasminogen activator vs. alfimeprase. AB - Plasmin-dependent thrombolytic agents are potentially prothrombotic and proinflammatory. Alfimeprase, a zinc-containing metalloproteinase, degrades fibrin directly and achieves thrombolysis independent of plasmin formation. This study examines the hypothesis that thrombolysis in the absence of plasmin generation results in improved myocardial salvage on reperfusion. The thrombolytic effects of recombinant tissue plasminogen activator [rt-PA; 0.022 mg/kg, 1/10 of which was administered as a loading dose; the rest (9/10) was infused over 60 min by intracoronary (ic) administration] or alfimeprase (0.5 mg/kg over 1 min ic) were evaluated in a canine model of arterial thrombosis involving electrolytic injury of the left circumflex (LCX) coronary artery. Both agents induced thrombolysis, with onset of reperfusion being more rapid after alfimeprase compared with rt-PA (1.5 +/- 0.6 vs. 10.1 +/- 2.1 min). In the absence of adjunctive therapy, time to reocclusion after alfimeprase was 3.2 +/- 0.5 min compared with 77.5 +/- 31.9 min with rt-PA. The glycoprotein IIb/IIIa platelet receptor antagonist CRL-42796 prolonged reperfusion time after thrombolysis with alfimeprase or rt-PA. The effect of each lytic agent on myocardial infarct size was examined in a separate group of dogs subjected to 60 min of LCX coronary artery ligation and 4 h of reperfusion. Myocardial infarct size, expressed as percentage of the risk region, was larger (32.16 +/- 3.95%) after rt-PA compared with alfimeprase (19.85 +/- 3.61%) or that of the saline control group (18.46 +/- 3.34%). rt-PA in contrast to alfimeprase, a direct acting fibrinolytic agent, is associated with an increase in myocyte reperfusion injury. PMID- 16243916 TI - Mesenchymal stem cell adhesion to cardiac microvascular endothelium: activators and mechanisms. AB - Circulating stem cells home within the myocardium, probably as the first step of a tissue regeneration process. This step requires adhesion to cardiac microvascular endothelium (CMVE). In this study, we studied mechanisms of adhesion between CMVE and mesenchymal stem cells (MSCs). Adhesion was studied in vitro and in vivo. Isolated 1,1'-dioctadecyl-3,3,3',3' tetramethylindocarbocyanine perchlorate-labeled rat MSCs were allowed to adhere to cultured CMVE in static and dynamic conditions. Either CMVE or MSCs were pretreated with cytokines [IL-1beta, IL-3, IL-6, stem cell factor, stromal cell derived factor-1, or TNF-alpha, 10 ng/ml]. Control or TNF-alpha-treated MSCs were injected intracavitarily in rat hearts in vivo. In baseline in vitro conditions, the number of MSCs that adhered to CMVE was highly dependent on the flow rate of the superfusing medium but remained significant at venous and capillary shear stress amplitudes. Activation of both CMVE and MSCs with TNF-alpha or IL-1beta before adhesion concentration dependently increased adhesion of MSCs at each studied level of shear stress. Consistently, in vivo, activation of MSCs with TNF alpha before injection significantly enhanced cardiac homing of MSCs. TNF-alpha induced adhesion could be completely blocked by pretreating either CMVE or MSCs with anti-VCAM-1 monoclonal antibodies but not by anti-ICAM-1 antibodies. Adhesion of circulating MSCs in the heart appears to be an endothelium-dependent process and is sensitive to modulation by activators of both MSCs and endothelium. Inflammation and the expression of VCAM-1 but not ICAM-1 on both cell types have a regulatory effect on MSC homing in the heart. PMID- 16243917 TI - Cyclic ADP ribose-mediated Ca2+ signaling in mediating endothelial nitric oxide production in bovine coronary arteries. AB - The present study tested the hypothesis that cyclic ADP ribose (cADPR) serves as a novel second messenger to mediate intracellular Ca2+ mobilization in coronary arterial endothelial cells (CAECs) and thereby contributes to endothelium dependent vasodilation. In isolated and perfused small bovine coronary arteries, bradykinin (BK)-induced concentration-dependent vasodilation was significantly attenuated by 8-bromo-cADPR (a cell-permeable cADPR antagonist), ryanodine (an antagonist of ryanodine receptors), or nicotinamide (an ADP-ribosyl cyclase inhibitor). By in situ simultaneously fluorescent monitoring, Ca2+ transient and nitric oxide (NO) levels in the intact coronary arterial endothelium preparation, 8-bromo-cADPR (30 microM), ryanodine (50 microM), and nicotinamide (6 mM) substantially attenuated BK (1 microM)-induced increase in intracellular [Ca2+] by 78%, 80%, and 74%, respectively, whereas these compounds significantly blocked BK-induced NO increase by about 80%, and inositol 1,4,5-trisphosphate receptor blockade with 2-aminethoxydiphenyl borate (50 microM) only blunted BK-induced Ca2+-NO signaling by about 30%. With the use of cADPR-cycling assay, it was found that inhibition of ADP-ribosyl cyclase by nicotinamide substantially blocked BK induced intracellular cADPR production. Furthermore, HPLC analysis showed that the conversion rate of beta-nicotinamide guanine dinucleotide into cyclic GDP ribose dramatically increased by stimulation with BK, which was blockable by nicotinamide. However, U-73122, a phospholipase C inhibitor, had no effect on this BK-induced increase in ADP-ribosyl cyclase activity for cADPR production. In conclusion, these results suggest that cADPR importantly contributes to BK- and A 23187-induced NO production and vasodilator response in coronary arteries through its Ca2+ signaling mechanism in CAECs. PMID- 16243918 TI - Contribution of p16INK4a and p21CIP1 pathways to induction of premature senescence of human endothelial cells: permissive role of p53. AB - We have previously found that nonenzymatically glycated collagen I (GC), mimicking diabetic microenvironment, can induce senescent phenotype in early passage human umbilical vein endothelial cells (HUVECs). In the present study, we explored the functional involvement of cell cycle checkpoint pathways in initiating GC-induced premature endothelial cell senescence. When compared with native collagen, early passage HUVECs showed increased p53, p21(CIP1) (p21), and p16(INK4a) (p16) mRNA expression after exposure to GC. Twenty-four hours after transfection of p16, p21, and p53-enhanced green fluorescent protein (EGFP) recombinant plasmids, HUVECs entered G(1)-phase cell cycle arrest. By days 3 and 5, HUVECs transfected with p16-EGFP showed an increased proportion of senescent cells, and this increase was more prominent in the GFP-positive cell population, which exhibited 68% of senescent cells. Transfection of p21 also induced senescence but only by day 5. Cotransfection of p16 and p21 showed no additive effect. Transfection of p21 or p53 induced apoptosis in HUVECs. Next, we suppressed endogenous p53, p21, p16, or retinoblastoma (Rb) gene expression through small interference RNA strategy and investigated their influence in p16- and p21-initiated endothelial cell senescence. Analysis indicated that suppression of p53 expression can abolish senescence induced by p16 overexpression. Paradoxically, this effect was not observed when p21 was suppressed. On the other hand, suppression of Rb eliminated senescence initiated by either p16 or p21 overexpression. In summary, the p53/p21 pathway is mainly responsible for GC-induced apoptosis, but the coordinated activation of the p53/p21 and p16 pathway is responsible for GC-induced endothelial cell senescence through a Rb-dependent mechanism. PMID- 16243919 TI - Adenosine and hypoxic dilation of rat coronary small arteries: roles of the ATP sensitive potassium channel, endothelium, and nitric oxide. AB - The aims of the study were to examine the roles of the ATP-sensitive potassium (K(ATP)) channel, the endothelium, and nitric oxide (NO) in the responses of rat coronary small arteries to adenosine and hypoxia. Segments of rat coronary vessel were investigated in vitro using pressure myography; all vessels studied developed stable spontaneous myogenic tone during equilibration. Glibenclamide (a K(ATP) channel inhibitor) reversed pinacidil but not 2-deoxyglucose-induced dilation. Both adenosine and hypoxia dilated the vessels, and glibenclamide did not reverse these responses. Endothelial removal or N(G)-nitro-L-arginine methyl ester (L-NAME) inhibited the dilation to adenosine by approximately 50%; subsequent addition of glibenclamide was without effect. Hypoxic dilation was completely inhibited by endothelium removal or L-NAME. We conclude that adenosine and hypoxia-induced dilation of rat coronary arteries does not appear to involve the K(ATP) channel. Adenosine-induced dilation is partially and hypoxic dilation is completely dependent on endothelium-derived NO. PMID- 16243920 TI - Cardiomyocyte function associated with hyperactivity and/or hypertension in genetic models of LV hypertrophy. AB - We examined cardiomyocyte intracellular calcium ([Ca2+]i) dynamics and sarcomere shortening dynamics in genetic rat models of left ventricular (LV) hypertrophy associated with or without hypertension (HT) and with or without hyperactive (HA) behavior. Previous selective breeding of the spontaneously hypertensive rat (SHR) strain, which is HA and HT, with the Wistar-Kyoto (WKY) rat strain, which is not hyperactive (NA) and not hypertensive (NT), has led to two unique strains: the WKHA strain, selected for HA and NT, and the WKHT strain, selected for NA and HT. Cardiomyocytes were isolated from young adult males and females of each strain, paced at 2, 3, and 4 Hz in 1.2 mM external Ca2+ concentration at 37 degrees C, and cardiomyocyte [Ca2+]i and sarcomere dynamics were recorded simultaneously. Under these conditions, LV cardiomyocyte systolic and diastolic [Ca2+]i dynamics and diastolic sarcomere dynamics in the WKHT were significantly enhanced compared with WKY controls, suggesting an underlying LV hypertrophic response that successfully compensated for HT in the absence of HA. LV cardiomyocyte [Ca2+]i dynamics in the WKHA and SHR were strikingly similar to each other and only slightly reduced compared with WKY. LV cardiomyocyte systolic and diastolic sarcomere dynamics, on the other hand, were significantly reduced in the SHR compare with WKHA and more so in male than in female SHR. We conclude from these data that HT alone is an insufficient descriptor of the cause of LV hypertrophy and diminished LV cardiomyocyte function in the SHR rat. These data further suggest that HA (augmented by male sex) in the SHR may interact with the HT state to initiate impaired cardiomyocyte function and thereby inhibit or undermine an otherwise compensatory response that may occur with HT in the absence of HA. PMID- 16243922 TI - Late coronary thrombosis in a sirolimus-eluting stent due to the lack of neointimal coverage. PMID- 16243921 TI - Extravascular pressure modulates responses of isolated rat coronary arteries to vasodilator, but not vasoconstrictor, stimuli. AB - The aims of the study were to investigate whether elevated extravascular pressure modulates responses of isolated rat coronary arteries to constrictor and dilator stimuli. Isolated segments of rat coronary artery were mounted in a modified pressure myograph system that allowed independent modulation of both intra- and extravascular pressures. The influence of elevated extravascular pressure on stable levels of myogenic tone and on responses to vasoconstrictor and vasodilator stimuli was investigated at constant overall transmural pressures. Stable levels of myogenic tone were independent of the relative levels of intra- and extravascular pressure, as were responses to depolarization and to addition of the thromboxane agonist U-46619. Elevating extravascular pressure, however, significantly reduced dilatory responses to introduction of intraluminal flow and to addition of endothelium-dependent and endothelium-independent vasodilatory agonists. These results support the notion that elevated extravascular pressure may attenuate responses of coronary arteries to a variety of dilatory stimuli. This finding may be of relevance to cardiac disorders associated with elevated ventricular pressures. PMID- 16243923 TI - Multimodality imaging of percutaneous closure of the left atrial appendage. PMID- 16243924 TI - Aortic dissection involving ostium of right coronary artery as the reason of myocardial infarction. PMID- 16243925 TI - Clinical vignette. Diagnosing acute myocarditis using cardiac MRI. PMID- 16243926 TI - Emergency endovascular repair of ruptured pseudo-aneurysm at the site of a corrected aortic coarctation. PMID- 16243951 TI - Response: on economic growth, business fluctuations, and health progress. PMID- 16243952 TI - Consequences of using different methods to assess cardiovascular risk in primary care. AB - BACKGROUND: There are two promising methods to assess cardiovascular risk: the Adult Treatment Panel III (ATPIII) and the Systematic Coronary Risk Evaluation (SCORE). The ATPIII calculates the 10-year risk of coronary events based on an adaptation of the original Framingham function. The SCORE chart is based on European studies and measures the absolute risk of cardiovascular mortality in the next 10 years. OBJECTIVE: To evaluate the clinical consequences of using different methods to calculate cardiovascular risk and different primary prevention guidelines. METHODS: A cross sectional study of 914 dyslipidemic patients from three primary health centres from Catalonia, Spain, was conducted. Outcome variables were the risk level according to the different equations (classical Framingham table by Anderson, ATPIII adapted Framingham table, and SCORE system), and candidates for lipid lowering treatment according to European and ATPIII guidelines. RESULTS: The proportion of high-risk patients according to the three equations and excluding diabetic patients was 13.5%, 11.4% and 7.1%, respectively, and 20.2%, 25.7% and 29.2%, respectively when including diabetic patients. The prevalence of candidates for lipid lowering treatment according to European guidelines and ATPIII guidelines were 28.8% and 39.3%, respectively. A 49% disagreement with a Kappa of -0.1, and a 37% disagreement with a Kappa of 0.08 were observed when comparing candidates identified for lipid lowering treatment and patients actually receiving that treatment, according to ATPIII and SCORE guidelines, respectively. CONCLUSION: Our results suggest important clinical and economic consequences when comparing European guidelines or ATPIII guidelines for the treatment of dyslipidemic patients in general practice. PMID- 16243953 TI - Testing a European set of indicators for the evaluation of the management of primary care practices. AB - BACKGROUND: Effective practice management is an important prerequisite for offering good clinical care. Internationally valid, reliable and feasible indicators and instruments are needed to describe and compare the management of primary care practices in Europe. OBJECTIVE: This paper describes development and evaluation of the European Practice Assessment instrument and indicators (Engels Y, Campbell S, Dautzenberg M et al. Developing a framework of, and quality indicators for, general practice management in Europe. Fam Pract 2005; 22(2): 215 22). METHODS: The study design was a validation and feasibility study set in 273 general practices in Austria, Belgium, France, Germany, Israel, The Netherlands, Slovenia, Switzerland and the UK. Use was made of a set of 62 valid quality indicators derived previously from an international Delphi procedure. The EPA instrument, based on this set of indicators, was used to collect data in the 273 practices. This instrument consists of self-completed questionnaires for doctors, staff managers and patients. In addition, there is an interview schedule for use by an outreach visitor, to be held with the lead GP or manager, and a visitor checklist. The instrument was analysed using expert review by the project partners, factor and reliability analyses, ANOVA analyses and by determining intraclass correlations. RESULTS: Fifty-seven indicators were found to be valid, feasible, reliable and discriminative in all participating countries. The instrument was able to determine differences in practice management within and between countries. All (but one) practices completed the assessment procedure. The data collection method appeared to be feasible, although some aspects can be improved. CONCLUSION: The EPA instrument provides feedback to practices that facilitates quality improvement and can compare primary care practices on a national and an international level. PMID- 16243954 TI - A multifaceted intervention according to the Audit Project Odense method improved secondary prevention of ischemic heart disease: a randomised controlled trial. AB - BACKGROUND: No single quality improvement instrument has proved consistently effective, but multifaceted interventions are believed to have the greatest impact. However, only little is known regarding what combinations are likely to be successful. OBJECTIVE: To evaluate the impact of a multifaceted intervention strategy combining GP registrations, outreach visits and feedback, targeting secondary prevention of ischemic heart disease in general practice. METHODS: A randomised controlled trial including 28 GPs in Ringkjobing County, Denmark. Half of the GPs received outreach visits and feedback on their prescribing of heart disease drugs. Evaluation was based on registration of consultations with patients suffering from ischemic heart disease. RESULTS: The intervention had a statistically significant impact on prescribing of lipid lowering drugs [odds ratio 1.59; 95% confidence interval (CI) 1.00 to 2.53] and acetylsalicylic acid (odds ratio 2.54; 95% CI 1.21 to 5.31). CONCLUSION: An intervention strategy combining outreach visits, feedback and GP registrations is a promising way of improving the quality of preventive treatment in general practice. PMID- 16243955 TI - Recovering from a heart attack: a qualitative study into lay experiences and the struggle to make lifestyle changes. AB - BACKGROUND: The adoption of healthy living advice by people with heart disease is known to be poor even in targeted interventions. Reasons for this can range from confusion about the seriousness of the condition to ineffectiveness in the form of advice and how it is conveyed. However, the social setting can be an important influence on lifestyle change. OBJECTIVES: To identify views and experiences of people recovering from myocardial infarction, specifically barriers to, and facilitators of, following advice about lifestyle change and maintenance. METHODS: Focus groups and interviews were undertaken with men and women discharged from hospital two/three years previously. A total of 53 people (35 men and 18 women) took part, recruited via a coronary care unit and patients' GPs. RESULTS: A major finding was participants' desires for long-term monitoring and support. While reported sources, form and content of coronary heart disease advice varied, most participants agreed that long-term follow up or back-up would be helpful, although what this should include and how it should be undertaken was not the same for all participants. This would fulfil needs such as: help in following lifestyle advice; sharing with people with similar experiences; regular contact with medical/health professionals (for confirmation of good heath and to ask questions); providing reassurance to other members of the patients' families. CONCLUSIONS: A long-term programme is needed incorporating mutual support and sharing with regular (not necessarily frequent) input from practitioners of information, advice and reassurance, as a support strategy for lifestyle change. PMID- 16243956 TI - Slow elimination of nonylphenol from rat intestine. AB - Nonylphenol, a possible endocrine disrupter, tends to persist in rat liver tissue after detoxification as a glucuronide conjugate by UDP-glucuronosyltransferase 2B1 expressed in the liver. In the intestine, however, the metabolism and dynamics of nonylphenol remain to be elucidated. The objectives of this study were to clarify the metabolism and excretion of nonylphenol having a long alkyl chain in the first barrier intestine and to estimate whether the nonylphenol alkyl chain governs the speed of excretion from intestinal tissue. Organ tissue glucuronidation activity toward alkylphenols (C2, C9) was investigated using microsomes prepared from intestinal tissue. To elucidate the elimination pathway of alkylphenols (C2, C4, C6, C9), a perfusion study was conducted on everted intestine. After oral administration (5 mg) of alkylphenols (C2, C9) to rats, gastrointestinal contents and related organ tissues (gastrointestinal tissue, liver, and kidney), blood, and urine were analyzed for alkylphenols (C2, C9) and glucuronides. The intestine showed strong glucuronidation activity toward alkylphenols (C2, C9). In everted intestinal assay, nonylphenol was glucuronidated within the intestinal wall, as was the case for other alkylphenols (C2, C4, C6), but nonylphenol-glucuronide was not excreted from intestinal tissue. Orally administered nonylphenol remained for long periods in gastrointestinal tissue as both the parent compound and glucuronide. The present study confirmed that intestinal tissue possesses an alkylphenol elimination system using UDP-glucuronosyltransferase; however, this system is impaired by the marginal transport of alkylphenol-glucuronide possessing long alkyl chain, such as nonylphenol. PMID- 16243957 TI - Transcriptional regulation of the PXR gene: identification and characterization of a functional peroxisome proliferator-activated receptor alpha binding site within the proximal promoter of PXR. AB - The pregnane X receptor (PXR, NR1I2) is widely regarded as a central factor in the body's response to changes in the fluxome, the overall metabolite profile in the body. PXR expression is regulated by a number of chemicals at the transcriptional level; the majority of these chemicals are ligands for PXR and substrates for PXR target genes. However, transcriptional activators of PXR, such as clofibrate, do not seem to be PXR ligands or substrates for its target genes. Understanding the molecular mechanisms underlying both these expected and, more importantly, unexpected transcriptional activations is central to fully understanding the roles of PXR in the human body. We have carried out an in silico analysis of the human PXR proximal promoter, identifying putative protein/DNA interaction sites within the 2 kilobases (kb) 5' to the putative transcription start site. These sites included several for liver-enriched transcription factors, such as the hepatic nuclear factors and CAAT-enhancer binding protein alpha, and chicken ovalbumin upstream promoter transcription factor, commensurate with the high expression of PXR in liver. Furthermore, we identified putative binding sites for a number of ligand-activated transcription factors, suggesting that these factors may regulate PXR gene expression. Further analysis of this regulatory region has shown that transcriptional activation of PXR by peroxisome proliferator-activated receptor alpha (PPARalpha) is via a binding site located approximately 1.3 kb upstream of the putative transcription start site, with ablation of this site preventing PPARalpha-mediated activation of PXR gene expression. We present a model of how regulation of PXR gene expression by ligand-activated transcription factors may play a central role in the body's response to xenobiotic exposure. PMID- 16243958 TI - Nuclear receptor expression in fetal and pediatric liver: correlation with CYP3A expression. AB - The mechanisms underlying interindividual variation and developmental changes in cytochrome P450 3A (CYP3A) expression and activity are not fully understood. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) methods were used to detect, during human fetal and pediatric development, mRNA expression of nuclear receptors involved in the regulation of CYP3A genes. Quantitative RT-PCR was conducted on RNA extracted from prenatal (n = 60, 76 days to 32 weeks estimated gestational age) and pediatric (n = 20, 4 days to 18 years of age) liver tissue with primers for nuclear receptors implicated in regulating CYP3A gene expression. Pregnane X receptor (PXR) and constitutive androstane receptor (CAR) were expressed at low (and highly variable) levels in pre- and neonatal liver relative to liver tissue derived from older children. CAR was expressed at higher levels relative to PXR in prenatal liver (757 +/- 480 molecules CAR/ng of RNA versus 271 +/- 190 molecules PXR/ng of RNA after correction for 18S rRNA). In contrast, mRNA expression of the heterodimer partner RXRalpha was less variable (33-fold) and did not differ appreciably between pre- and postnatal liver samples (219 +/- 101 molecules/ng of RNA prenatal versus 253 +/- 232 molecules/ng of RNA postnatal). Expression of HNF4alpha1 mRNA was similar to that of RXRalpha. Log CYP3A7 mRNA expression was significantly correlated with PXR (r2 = 0.372) and CAR (r2 = 0.380) mRNA in fetal liver, but associations were weaker than those observed with CYP3A4 mRNA in postnatal liver (r2 = 0.610 and 0.723 for PXR and CAR, respectively). In conclusion, nuclear receptor mRNA expression demonstrates considerable interindividual variability in human fetal and pediatric liver and is significantly correlated with CYP3A expression. PMID- 16243959 TI - In vitro metabolism of naphthalene by human liver microsomal cytochrome P450 enzymes. AB - The polycyclic aromatic hydrocarbon naphthalene is an environmental pollutant, a component of jet fuel, and, since 2000, has been reclassified as a potential human carcinogen. Few studies of the in vitro human metabolism of naphthalene are available, and these focus primarily on lung metabolism. The current studies were performed to characterize naphthalene metabolism by human cytochromes P450. Naphthalene metabolites from pooled human liver microsomes (pHLMs) were trans-1,2 dihydro-1,2-naphthalenediol (dihydrodiol), 1-naphthol, and 2-naphthol. Metabolite production generated Km values of 23, 40, and 116 microM And Vmax values of 2860, 268, and 22 pmol/mg protein/min, respectively. P450 isoform screening of naphthalene metabolism identified CYP1A2 as the most efficient isoform for producing dihydrodiol and 1-naphthol, and CYP3A4 as the most effective for 2 naphthol production. Metabolism of the primary metabolites of naphthalene was also studied to identify secondary metabolites. Whereas 2-naphthol was readily metabolized by pHLMs to produce 2,6- and 1,7-dihydroxynaphthalene, dihydrodiol and 1-naphthol were inefficient substrates for pHLMs. A series of human p450 isoforms was used to further explore the metabolism of dihydrodiol and 1 naphthol. 1,4-Naphthoquinone and four minor unknown metabolites from 1-naphthol were observed, and CYP1A2 and 2D6*1 were identified as the most active isoforms for the production of 1,4-naphthoquinone. Dihydrodiol was metabolized by P450 isoforms to three minor unidentified metabolites with CYP3A4 and CYP2A6 having the greatest activity toward this substrate. The metabolism of dihydrodiol by P450 isoforms was lower than that of 1-naphthol. These studies identify primary and secondary metabolites of naphthalene produced by pHLMs and P450 isoforms. PMID- 16243960 TI - Transcriptional regulation of the NAD(P)H:quinone oxidoreductase 1 and glutathione S-transferase ya genes by mercury, lead, and copper. AB - Recently, we demonstrated the ability of heavy metals, particularly Hg2+, Pb2+, and Cu2+, to differentially modulate in Hepa 1c1c7 cells the expression of the phase II xenobiotic metabolizing enzymes NAD(P)H:quinone oxidoreductase 1 (Nqo1) and glutathione S-transferase subunit Ya (Gst ya) genes, yet the mechanisms involved remain unknown. To investigate the molecular mechanisms involved in the regulation of Nqo1 and Gst ya genes by heavy metals, Hepa 1c1c7 cells were treated with Hg2+, Pb2+, or Cu2+ in the presence and absence of 2,3,7,8 tetrachlorodibenzo-p-dioxin, a potent inducer of Nqo1, Gst ya, and Cyp1a1 genes. Analysis of the time-dependent effect of heavy metals revealed that Hg2+ and Pb2+ increased whereas Cu2+ inhibited the constitutive and inducible expression of Nqo1 and Gst ya mRNAs in a time-dependent manner. The RNA synthesis inhibitor actinomycin D significantly inhibited the Nqo1 and Gst ya mRNA induction in response to metals, indicating a requirement of de novo RNA synthesis. The protein synthesis inhibitor cycloheximide significantly inhibited metal-mediated induction of Nqo1 and Gst ya mRNAs, which coincided with a decrease in the nuclear factor erythroid 2-related factor 2 (Nrf2) protein expression, implying the requirement of Nrf2 protein synthesis for the induction of these genes. Furthermore, inhibition of Nrf2 protein degradation by carbobenzoxy-L-leucyl-L leucyl-leucinal (MG-132), a 26S proteasome inhibitor, significantly reversed the cycloheximide-mediated inhibition of Nqo1 and Gst ya mRNAs, which coincided with an increase in the expression of Nrf2, confirming that a transcriptional mechanism is involved. Nqo1 and Gst ya mRNA and protein decay experiments revealed lack of post-transcriptional and post-translational mechanisms. This is the first demonstration that heavy metals regulate the expression of Nqo1 and Gst ya genes through a transcriptional mechanism. PMID- 16243961 TI - M2 and M3 muscarinic receptor activation of urinary bladder contractile signal transduction. I. Normal rat bladder. AB - The muscarinic receptor subtype-activated signal transduction mechanisms mediating rat urinary bladder contraction are incompletely understood. M(3) mediates normal rat bladder contractions; however, the M(2) receptor subtype has a more dominant role in contractions of the hypertrophied bladder. Normal bladder muscle strips were exposed to inhibitors of enzymes thought to be involved in signal transduction in vitro followed by a single cumulative concentration response curve to the muscarinic receptor agonist carbachol. The outcome measures were the maximal contraction, the potency of carbachol, and the affinity of the M(3) -selective antimuscarinic agent darifenacin for inhibition of contraction. Inhibition of phosphoinositide-specific phospholipase C (PI-PLC) with 1-O octadecyl-2-O-methyl-sn-glycero-3-phosphorylcholine (ET-18-OCH(3)) reduces carbachol potency and reduces darifenacin affinity, whereas inhibition of phosphatidyl choline-specific phospholipase C (PC-PLC) with O tricyclo[5.2.1.02,6]dec-9-yl dithiocarbonate potassium salt (D609) attenuates the carbachol maximal contraction. Inhibition of rho kinase with (R)-(+)-trans-4-(1 aminoethyl)-N-(4-pyridyl)cyclohexanecarboxamide dihydrochloride (Y-27632) reduces carbachol potency and increases darifenacin affinity. Inhibition of rho kinase, protein kinase A (PKA), and protein kinase G (PKG) with 1-(5 isoquinolinesulfonyl)-homopiperazine.HCl (HA-1077) reduces the carbachol maximal contraction, carbachol potency, and darifenacin affinity. Inhibition of protein kinase C (PKC) with chelerythrine increases darifenacin affinity, whereas inhibition of rho kinase, PKA, PKG, and PKC with 1-(5-isoquinolinesulfonyl)-2 methylpiperazine.2HCl (H7) reduces the carbachol maximum and carbachol potency while increasing darifenacin affinity. Inhibition of rho kinase, PKA, and PKG with N-[2-(p-bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide.2HCl (H89) reduces carbachol maximum and carbachol potency. Both the M(2) and the M(3) receptor subtype are involved in normal rat bladder contractions. The M(3)subtype seems to mediate contraction by activation of PI-PLC, PC-PLC, and PKA, whereas the M(2) signal transduction cascade may include activation of rho kinase, PKC, and an additional contractile signal transduction mechanism independent of rho kinase or PKC. PMID- 16243962 TI - M2 and M3 muscarinic receptor activation of urinary bladder contractile signal transduction. II. Denervated rat bladder. AB - Normal rat bladder contractions are mediated by the M(3) muscarinic receptor subtype. The M(2) receptor subtype mediates contractions of the denervated, hypertrophied bladder. This study determined signal transduction mechanisms mediating contraction of the denervated rat bladder. Denervated bladder muscle strips were exposed to inhibitors of enzymes thought to be involved in signal transduction in vitro followed by a cumulative carbachol concentration-response curve. Outcome measures were the maximal contraction, the potency of carbachol, and the affinity of darifenacin for inhibition of contraction. Inhibition of phosphoinositide-specific phospholipase C (PI-PLC) with 1-O-octadecyl-2-O-methyl sn-glycero-3-phosphorylcholine (ET-18-OCH(3)) has no effect on denervated bladder contractions, whereas inhibition of phosphatidyl choline-specific phospholipase C (PC-PLC) with O-tricyclo[5.2.1.02,6]dec-9-yl dithiocarbonate potassium salt (D609) attenuates the carbachol maximum and potency. Inhibition of rho kinase with (R)-(+)-trans-4-(1-aminoethyl)-N-(4-pyridyl)cyclohexanecarboxamide dihydrochloride (Y-27632) reduces carbachol maximum, carbachol potency, and increases darifenacin affinity. Inhibition of rho kinase, protein kinase A (PKA), and protein kinase G (PKG) with 1-(5-isoquinolinesulfonyl)-homopiperazine.HCl (HA 1077) reduces the carbachol maximum and potency. Inhibition of PKC with chelerythrine increases darifenacin affinity, whereas inhibition of rho kinase, PKA, PKG, and protein kinase C (PKC) with 1-(5-isoquinolinesulfonyl)-2 methylpiperazine.2HCl (H7) reduces the carbachol potency while increasing darifenacin affinity. Inhibition of rho kinase, PKA, and PKG with N-[2-(p bromocinnamylamino)ethyl]-5-isoquinolinesulfonamide.2HCl (H89) increases darifenacin affinity. This study demonstrates that different signal transduction mechanisms mediate the contractile response in the denervated rat bladder than in normal rat bladder. In normal rat bladder, PI-PLC and PC-PLC mediate the contraction, but in denervated bladder only PC-PLC is involved. In the denervated bladder, the rho kinase pathway is more dominant than in normal bladders. PKA seems to mediate a contractile response in normal bladders, whereas it seems to inhibit contraction in denervated bladders. PMID- 16243963 TI - In vivo cardiac electrophysiologic effects of a novel diphenylphosphine oxide IKur blocker, (2-Isopropyl-5-methylcyclohexyl) diphenylphosphine oxide, in rat and nonhuman primate. AB - The voltage-gated potassium channel, Kv1.5, which underlies the ultrarapid delayed rectifier current, I(Kur), is reported to be enriched in human atrium versus ventricle, and has been proposed as a target for novel atrial antiarrhythmic therapy. The administration of the novel I(Kur) blocker (2 isopropyl-5-methyl-cyclohexyl) diphenylphosphine oxide (DPO-1) (0.06, 0.2, and 0.6 mg/kg/min i.v. x 20 min; total doses 1.2, 4.0, and 12.0 mg/kg, respectively) to rat, which exhibits I(Kur) in both atria and ventricle, elicited significant, dose-dependent increases in atrial and ventricular refractory period (9-42%) at all doses tested, with no changes in cardiac rate or indices of cardiac conduction. Plasma levels achieved in rat at the end of the three infusions were 1.1, 4.1, and 7.7 microM. Reverse transcription-polymerase chain reaction analysis of African green monkey atria and ventricle demonstrated an atrial preferential distribution of Kv1.5 transcript. The administration of DPO-1 (1.0, 3.0, and 10.0 mg/kg i.v.; 5-min infusions) to African green monkey elicited significant increases in atrial refractoriness (approximately 15% increase at the 10.0 mg/kg dose), with no change in ventricular refractory period, ECG intervals, heart rate, or blood pressure. Plasma levels of DPO-1 achieved in African green monkey were 0.58, 1.12, and 5.43 microM. The concordance of effect of DPO-1 on myocardial refractoriness with distribution of Kv1.5 in these two species is consistent with the I(Kur) selectivity of DPO-1 in vivo. Moreover, the selective increase in atrial refractoriness in primate supports the concept of I(Kur) blockade as an approach for the development of atrial-specific antiarrhythmic agents. PMID- 16243964 TI - The effect of select nutrients on serum high-density lipoprotein cholesterol and apolipoprotein A-I levels. AB - One of the factors contributing to the increased risk of developing premature atherosclerosis is low plasma concentrations of high-density lipoprotein (HDL) cholesterol (HDLc). Multiple potential mechanisms account for the cardioprotective effects of HDL and its main protein apolipoprotein A-I (apo A I). The low plasma concentrations of HDL could be the result of increased fractional clearance and reduced expression of apo A-I. To this end, nutrients play an important role in modulating the fractional clearance rate, as well as the rate of apo A-I gene expression. Because medical nutrition therapy constitutes the cornerstone of management of dyslipidemias, it is essential to understand the mechanisms underlying the changes in HDL level in response to alterations in dietary intake. In this review, we will discuss the effect of select nutrients on serum HDLc and apo A-I levels. Specifically, we will review the literature on the effect of carbohydrates, fatty acids, and ketones, as well as some of the nutrient-related metabolites, such as glucosamine and the prostanoids, on apo A-I gene expression. Because there are multiple mechanisms involved in the regulation of serum HDLc levels, changes in gene transcription do not necessarily correlate with clinical observations on serum levels of HDLc. PMID- 16243965 TI - Structural activation pathways from dynamic olfactory receptor-odorant interactions. AB - We have simulated an odor ligand's dynamic behavior in the binding region of an olfactory receptor (OR). Our short timescale computational studies (up to 200 ps) have helped identify unprecedented postdocking ligand behavior of ligands. From in vacuo molecular dynamics simulations of interactions between models of rat OR I7 and 10 aldehyde ligands, we have identified a dissociative pathway along which the ligand exits and enters the OR-binding pocket--a transit event. The ligand's transit through the receptor's binding region may mark the beginning of a signal transduction cascade leading to odor recognition. We have graphically traced the rotameric changes in key OR amino acid side chains during the transit. Our results have helped substantiate or refute previously held notions of amino acid contribution to ligand stability in the binding pocket. Our observations of ligand activity when compared to those of experimental (electroolfactogram response) OR-activation studies provide a view to predicting the stability of ligands in the binding pocket as a precursor to OR activation by the ligand. PMID- 16243966 TI - Taste cell responses in the frog are modulated by parasympathetic efferent nerve fibers. AB - We studied the anatomical properties of parasympathetic postganglionic neurons in the frog tongue and their modulatory effects on taste cell responses. Most of the parasympathetic ganglion cell bodies in the tongue were found in extremely small nerve bundles running near the fungiform papillae, which originate from the lingual branches of the glossopharyngeal (GP) nerve. The density of parasympathetic postganglionic neurons in the tongue was 8000-11,000/mm(3) of the extremely small nerve bundle. The mean major axis of parasympathetic ganglion cell bodies was 21 microm, and the mean length of parasympathetic postganglionic neurons was 1.45 mm. Electrical stimulation at 30 Hz of either the GP nerve or the papillary nerve produced slow hyperpolarizing potentials (HPs) in taste cells. After nicotinic acetyl choline receptors on the parasympathetic ganglion cells in the tongue had been blocked by intravenous (i.v.) injection of D tubocurarine (1 mg/kg), stimulation of the GP nerve did not induce any slow HPs in taste cells but that of the papillary nerve did. A further i.v. injection of a substance P NK-1 antagonist, L-703,606, blocked the slow HPs induced by the papillary nerve stimulation. This suggests that the parasympathetic postganglionic efferent fibers innervate taste cells and are related to a generation of the slow HPs and that substance P is released from the parasympathetic postganglionic axon terminals. When the resting membrane potential of a taste cell was hyperpolarized by a prolonged slow HP, the gustatory receptor potentials for NaCl and sugar stimuli were enhanced in amplitude, but those for quinine-HCl and acetic acid stimuli remained unchanged. It is concluded that frog taste cell responses are modulated by activities of parasympathetic postganglionic efferent fibers innervating these cells. PMID- 16243967 TI - In situ modification of herbivore-induced plant odors: a novel approach to study the attractiveness of volatile organic compounds to parasitic wasps. AB - Many parasitic wasps (parasitoids) exploit volatile organic compounds (VOCs) emitted by herbivore-infested plants in order to locate their hosts, but it remains largely unknown which specific compounds within the volatile blends elicit the attractiveness to parasitoids. One way of studying the importance of specific VOCs is to test the attractiveness of odor blends from which certain compounds have been emitted. We used this approach by testing the attraction of naive and experienced females of the two parasitoids Cotesia marginiventris and Microplitis rufiventris to partially altered volatile blends of maize seedlings (Zea mays var. Delprim) infested with Spodoptera littoralis larvae. Adsorbing filter tubes containing carbotrap-C or silica were installed in a four-arm olfactometer between the odor source vessels and the arms of the olfactometer. The blends breaking through were tested for chemical composition and attractiveness to the wasps. Carbotrap-C adsorbed most of the sesquiterpenes, but the breakthrough blend remained attractive to naive C. marginiventris females. Silica adsorbed only some of the more polar VOCs, but this essentially eliminated all attractiveness to naive C. marginiventris, implying that among the adsorbed compounds there are some that play key roles in the attraction. Unlike C. marginiventris, M. rufiventris was still attracted to the latter blend, showing that parasitoids with a comparable biology may employ different strategies in their use of plant-provided cues to locate hosts. Results from similar experiments with modified odor blends of caterpillar-infested cowpea (Vigna unguiculata) indicate that key VOCs in different plant species vary greatly in quality and/or quantity. Finally, experienced wasps were more strongly attracted to a specific blend after they perceived the blend while ovipositing in a host. Considering the high number of distinct adsorbing materials available today, this in situ modification of complex volatile blends provides a new and promising approach pinpointing on key attractants within these blends. Advantages and disadvantages compared to other approaches are discussed. PMID- 16243968 TI - Quantitative high-throughput analysis of DNA methylation patterns by base specific cleavage and mass spectrometry. AB - Methylation is one of the major epigenetic processes pivotal to our understanding of carcinogenesis. It is now widely accepted that there is a relationship between DNA methylation, chromatin structure, and human malignancies. DNA methylation is potentially an important clinical marker in cancer molecular diagnostics. Understanding epigenetic modifications in their biological context involves several aspects of DNA methylation analysis. These aspects include the de novo discovery of differentially methylated genes, the analysis of methylation patterns, and the determination of differences in the degree of methylation. Here we present a previously uncharacterized method for high-throughput DNA methylation analysis that utilizes MALDI-TOF mass spectrometry (MS) analysis of base-specifically cleaved amplification products. We use the IGF2/H19 region to show that a single base-specific cleavage reaction is sufficient to discover methylation sites and to determine methylation ratios within a selected target region. A combination of cleavage reactions enables the complete evaluation of all relevant aspects of DNA methylation, with most CpGs represented in multiple reactions. We successfully applied this technology under high-throughput conditions to quantitatively assess methylation differences between normal and neoplastic lung cancer tissue samples from 48 patients in 47 genes and demonstrate that the quantitative methylation results allow accurate classification of samples according to their histopathology. PMID- 16243969 TI - Support from the relationship of genetic and geographic distance in human populations for a serial founder effect originating in Africa. AB - Equilibrium models of isolation by distance predict an increase in genetic differentiation with geographic distance. Here we find a linear relationship between genetic and geographic distance in a worldwide sample of human populations, with major deviations from the fitted line explicable by admixture or extreme isolation. A close relationship is shown to exist between the correlation of geographic distance and genetic differentiation (as measured by F(ST)) and the geographic pattern of heterozygosity across populations. Considering a worldwide set of geographic locations as possible sources of the human expansion, we find that heterozygosities in the globally distributed populations of the data set are best explained by an expansion originating in Africa and that no geographic origin outside of Africa accounts as well for the observed patterns of genetic diversity. Although the relationship between F(ST) and geographic distance has been interpreted in the past as the result of an equilibrium model of drift and dispersal, simulation shows that the geographic pattern of heterozygosities in this data set is consistent with a model of a serial founder effect starting at a single origin. Given this serial-founder scenario, the relationship between genetic and geographic distance allows us to derive bounds for the effects of drift and natural selection on human genetic variation. PMID- 16243971 TI - Scientific issues in the design of metrics for inclusion of oxides of nitrogen in global climate agreements. AB - The Kyoto Protocol seeks to limit emissions of various greenhouse gases but excludes short-lived species and their precursors even though they cause a significant climate forcing. We explore the difficulties that are faced when designing metrics to compare the climate impact of emissions of oxides of nitrogen (NO(x)) with other emissions. There are two dimensions to this difficulty. The first concerns the definition of a metric that satisfactorily accounts for its climate impact. NO(x) emissions increase tropospheric ozone, but this increase and the resulting climate forcing depend strongly on the location of the emissions, with low-latitude emissions having a larger impact. NO(x) emissions also decrease methane concentrations, causing a global-mean radiative forcing similar in size but opposite in sign to the ozone forcing. The second dimension of difficulty concerns the intermodel differences in the values of computed metrics. We explore the use of indicators that could lead to metrics that, instead of using global-mean inputs, are computed locally and then averaged globally. These local metrics may depend less on cancellation in the global mean; the possibilities presented here seem more robust to model uncertainty, although their applicability depends on the poorly known relationship between local climate change and its societal/ecological impact. If it becomes a political imperative to include NO(x) emissions in future climate agreements, policy makers will be faced with difficult choices in selecting an appropriate metric. PMID- 16243970 TI - The highly conserved cardiac glycoside binding site of Na,K-ATPase plays a role in blood pressure regulation. AB - The Na,K-ATPase contains a binding site for cardiac glycosides, such as ouabain, digoxin, and digitoxin, which is highly conserved among species ranging from Drosophila to humans. Although advantage has been taken of this site to treat congestive heart failure with drugs such as digoxin, it is unknown whether this site has a natural function in vivo. Here we show that this site plays an important role in the regulation of blood pressure, and it specifically mediates adrenocorticotropic hormone (ACTH)-induced hypertension in mice. We used genetically engineered mice in which the Na,K-ATPase alpha2 isoform, which is normally sensitive to cardiac glycosides, was made resistant to these compounds. Chronic administration of ACTH caused hypertension in WT mice but not in mice with an ouabain-resistant alpha2 isoform of Na,K-ATPase. This finding demonstrates that the cardiac glycoside binding site of the Na,K-ATPase plays an important role in blood pressure regulation, most likely by responding to a naturally occurring ligand. Because the alpha1 isoform is sensitive to cardiac glycosides in humans, we developed mice in which the naturally occurring ouabain resistant alpha1 isoform was made ouabain-sensitive. Mice with the ouabain sensitive "human-like" alpha1 isoform and an ouabain-resistant alpha2 isoform developed ACTH-induced hypertension to greater extent than WT animals. This result indicates that the cardiac glycoside binding site of the alpha1 isoform can also mediate ACTH-induced hypertension. Taken together these results demonstrate that the cardiac glycoside binding site of the alpha isoforms of the Na,K-ATPase have a physiological function and supports the hypothesis for a role of the endogenous cardiac glycosides. PMID- 16243972 TI - Light scattered by model phantom bacteria reveals molecular interactions at their surface. AB - Testing molecular interactions is an ubiquitous need in modern biology and molecular medicine. Here, we present a qualitative and quantitative method rooted in the basic properties of the scattering of light, enabling detailed measurement of ligand-receptor interactions occurring on the surface of colloids. The key factor is the use of receptor-coated nanospheres matched in refractive index with water and therefore optically undetectable ("phantom") when not involved in adhesion processes. At the occurrence of ligand binding at the receptor sites, optically unmatched material adsorbs on the nanoparticle surface, giving rise to an increment in their scattering cross section up to a maximum corresponding to saturated binding sites. The analysis of the scattering growth pattern enables extracting the binding affinity. This label-free method has been assessed through the determination of the binding constant of the antibiotic vancomycin with the tripeptide l-Lys-d-Ala-d-Ala and of the vancomycin dimerization constant. We shed light on the role of chelate effect and molecular hindrance in the activity of this glycopeptide. PMID- 16243973 TI - Inhibitors of histone deacetylases target the Rb-E2F1 pathway for apoptosis induction through activation of proapoptotic protein Bim. AB - Inhibitors of histone deacetylases (HDACIs) are a new generation of anticancer agents that selectively kill tumor cells. However, the molecular basis for their tumor selectivity is not well understood. We investigated the effects of HDACIs on the oncogenic Rb-E2F1 pathway, which is frequently deregulated in human cancers. Here, we report that cancer cells with elevated E2F1 activity, caused either by enforced E2F1 expression, or by E1A oncogene expression, are highly susceptible to HDACI-induced cell death. This E2F1-mediated apoptosis is neither p53- nor p73-dependent but proceeds through selective induction of proapoptotic BH3-only protein Bim. We show that Bim is a direct target of E2F1 and that HDAC inhibition promotes the recruitment of E2F1 to the Bim promoter. Moreover, silencing of Bim by specific small interfering RNA (siRNA) effectively abolishes the E2F1-mediated cell death sensitization to HDACIs. These findings suggest that the oncogenic E2F1 pathway participates in HDACIs-induced apoptosis in cancer cells and underscore the importance of Bim as a key mediator of oncogene-induced apoptosis. Our study provides an important insight into the molecular mechanism of tumor selectivity of HDACIs and predicts that, clinically, HDACIs will be more effective in tumors with high E2F1 activity. PMID- 16243975 TI - Measurement of the contributions of 1D and 3D pathways to the translocation of a protein along DNA. AB - Proteins that act at specific DNA sequences bind DNA randomly and then translocate to the target site. The translocation is often ascribed to the protein sliding along the DNA while maintaining continuous contact with it. Proteins also can move on DNA by multiple cycles of dissociation/reassociation within the same chain. To distinguish these pathways, a strategy was developed to analyze protein motion between DNA sites. The strategy reveals whether the protein maintains contact with the DNA as it transfers from one site to another by sliding or whether it loses contact by a dissociation/reassociation step. In reactions at low salt, the test protein stayed on the DNA as it traveled between sites, but only when the sites were <50 bp apart. Transfers of >30 bp at in vivo salt, and over distances of >50 bp at any salt, always included at least one dissociation step. Hence, for this enzyme, 1D sliding operates only over short distances at low salt, and 3D dissociation/reassociation is its main mode of translocation. PMID- 16243974 TI - A fully dissociated compound of plant origin for inflammatory gene repression. AB - The identification of selective glucocorticoid receptor (GR) modifiers, which separate transactivation and transrepression properties, represents an important research goal for steroid pharmacology. Although the gene-activating properties of GR are mainly associated with undesirable side effects, its negative interference with the activity of transcription factors, such as NF-kappaB, greatly contributes to its antiinflammatory and immune-suppressive capacities. In the present study, we found that Compound A (CpdA), a plant-derived phenyl aziridine precursor, although not belonging to the steroidal class of GR-binding ligands, does mediate gene-inhibitory effects by activating GR. We demonstrate that CpdA exerts an antiinflammatory potential by down-modulating TNF-induced proinflammatory gene expression, such as IL-6 and E-selectin, but, interestingly, does not at all enhance glucocorticoid response element-driven genes or induce GR binding to glucocorticoid response element-dependent genes in vivo. We further show that the specific gene-repressive effect of CpdA depends on the presence of functional GR, displaying a differential phosphorylation status with CpdA as compared with dexamethasone treatment. The antiinflammatory mechanism involves both a reduction of the in vivo DNA-binding activity of p65 as well as an interference with the transactivation potential of NF-kappaB. Finally, we present evidence that CpdA is as effective as dexamethasone in counteracting acute inflammation in vivo and does not cause a hyperglycemic side effect. Taken together, this compound may be a lead compound of a class of antiinflammatory agents with fully dissociated properties and might thus hold great potential for therapeutic use. PMID- 16243978 TI - Isoflurane prevents platelets from enhancing neutrophil-induced coronary endothelial dysfunction. AB - We evaluated whether platelets can enhance polymorphonuclear neutrophil-induced coronary endothelial dysfunction, and, after observing this, whether isoflurane can prevent the effect. Neutrophils, coronary artery segments, and platelets were obtained from 25 healthy dogs. Coronary artery rings were exposed to neutrophils activated with platelet-activating factor (1.0 microM), and after washing and preconstriction with U46619, were evaluated for concentration-related responses to acetylcholine, an endothelium-dependent vasorelaxing drug. Superoxide production by activated neutrophils was measured spectrophotometrically. Adherence of the activated neutrophils to the endothelium of coronary segments was assessed by direct counting of neutrophils labeled with fluorescent dye. Measurements were performed in absence and presence of isoflurane (1 minimum alveolar concentration) both with and without platelets. The presence of platelets enhanced the neutrophil-induced rightward shift in the concentration vasorelaxation response curve to acetylcholine (the concentration of acetylcholine required to elicit 50% of maximal relaxation (-log M) was increased from 6.78 +/- 0.7 to 5.26 +/- 0.6), and it increased superoxide oxide production from 45.0 +/- 4.2 to 54.3 +/- 4.2 nM O2-/5 x 10(6) neutrophils and adherence of activated neutrophils from 204 +/- 10 to 268 +/- 5 neutrophils/mm2. Isoflurane abolished these effects of platelets. In conclusion, platelets enhanced the ability of neutrophils to cause coronary endothelial dysfunction. This effect was prevented by isoflurane. This may be attributable to an inhibitory action on superoxide production by the neutrophils leading to reduced expression of endothelial adhesion molecules and, in turn, reduced neutrophil adherence. PMID- 16243976 TI - Interferon regulatory factor 4 negatively regulates the production of proinflammatory cytokines by macrophages in response to LPS. AB - A member of the IFN regulatory factor (IRF) family of transcription factors, IRF 4 is expressed in lymphocytes and macrophage/dendritic cells. Studies using IRF-4 deficient mice have revealed the critical roles of IRF-4 in lymphocyte responses. However, the role of IRF-4 in innate immune responses is not clearly understood. Here, we demonstrate that IRF-4 negatively regulates the production of proinflammatory cytokines by macrophages in response to Toll-like receptor (TLR) stimulation. Mice lacking IRF-4 are sensitive to LPS-induced shock, and their macrophages produce high levels of proinflammatory cytokines, including TNF-alpha and IL-6, in response to TLR ligands. The inhibitory role of IRF-4 in response to TLR stimulation was confirmed by the down-regulation of IRF-4 expression in normal macrophages by using the small interfering RNA technique and by the overexpression of IRF-4 in macrophage line RAW264.7. Activation of the important signaling pathways for cytokine production, NF-kappaB and JNK (c-Jun N-terminal kinase), was enhanced after LPS stimulation in IRF-4(-/-) macrophages. These results imply that IRF-4 negatively regulates TLR signaling and is inhibitory to the production of proinflammatory cytokines in response to TLR stimulation. PMID- 16243977 TI - The influence of mitochondrial KATP-channels in the cardioprotection of preconditioning and postconditioning by sevoflurane in the rat in vivo. AB - Volatile anesthetics induce myocardial preconditioning and can also protect the heart when given at the onset of reperfusion-a practice recently termed "postconditioning." We investigated the role of mitochondrial KATP (mKATP) channels in sevoflurane-induced cardioprotection for both preconditioning and postconditioning alone and whether there is a synergistic effect of both. Rats were subjected to 25 min of coronary artery occlusion followed by 120 min of reperfusion. Infarct size was determined by triphenyltetrazolium staining. The following protocols were used: 1) preconditioning (S-Pre, n = 10, achieved by 2 periods of 5 min sevoflurane administration (1 MAC) followed by 10 min of washout); 2) sevoflurane postconditioning (1 MAC of sevoflurane given for 2 min at the beginning of reperfusion; S-Post, n = 10); 3) administration before and after ischemia (S-Pre + S-Post, n = 10). Protocols 1-3 were repeated in the presence of 5-hydroxydecanoate (5HD), a specific mKATP-channel-blocker (S-Pre + S Post + 5HD, S-Pre + 5HD: n = 10; S-Post + 5HD: n = 9). Nine rats served as untreated controls (CON) or received 5HD alone (5HD, n = 10). Both S-Pre (23% +/- 13% of the area at risk, mean +/- sd) and S-Post (18% +/- 5%) reduced infarct size compared with CON (49% +/- 11%, both P < 0.05). S-Pre + S-Post resulted in a larger reduction of infarct size (12% +/- 5%, P = 0.054 versus S-Pre) compared with administration before or after ischemia alone. 5HD diminished the protection in all three sevoflurane treated groups (S-Pre + 5HD, 35% +/- 12%; S-Post + 5HD, 44% +/- 12%; S-Pre + S-Post + 5HD, 46% +/- 14%;) but given alone had no effect on infarct size (41% +/- 13%). Sevoflurane preconditioning and postconditioning protects against myocardial ischemia-reperfusion injury. The combination of preconditioning and postconditioning provides additive cardioprotection and is mediated, at least in part, by mKATP-channels. PMID- 16243979 TI - Myocardial systolic function increases during positive pressure lung inflation. AB - Lung inflation with positive airway pressure may have rapid and dynamic effects on myocardial contractile function. We designed this study to assess the magnitude and time to onset of myocardial function changes during the initiation of single positive pressure lung inflation at clinically relevant inflation pressures. In 8 anesthetized 40-kg pigs, left ventricular pressures and volumes were measured directly (conductance volumetry). A 15 cm H2O airway pressure plateau with lung inflation (PPLI-15) was performed, and 2 single beats from that sequence, one from resting apnea at zero airway pressure and the second from the point when the lungs were first maximally inflated, were selected for analysis. Systolic function variables for zero airway pressure and PPLI-15 were analyzed. Systolic elastance, derived from bilinear time-varying elastance curves, increased approximately 15% during PPLI-15 from zero airway pressure. This agreed with other systolic function variables that identified an increase in left ventricular contractile function for the lung inflation beat. Serial measurements of myocardial function should be conducted with constant airway pressure and lung inflation conditions. PMID- 16243980 TI - Reactive oxygen species as mediators of cardiac injury and protection: the relevance to anesthesia practice. AB - Reactive oxygen species (ROS) are central to cardiac ischemic and reperfusion injury. They contribute to myocardial stunning, infarction and apoptosis, and possibly to the genesis of arrhythmias. Multiple laboratory studies and clinical trials have evaluated the use of scavengers of ROS to protect the heart from the effects of ischemia and reperfusion. Generally, studies in animal models have shown such effects. Clinical trials have also shown protective effects of scavengers, but whether this protection confers meaningful clinical benefits is uncertain. Several IV anesthetic drugs act as ROS scavengers. In contrast, volatile anesthetics have recently been demonstrated to generate ROS in the heart, most likely because of inhibitory effects on cardiac mitochondria. ROS are involved in the signaling cascade for cardioprotection induced by brief exposure to a volatile anesthetic (termed "anesthetic preconditioning"). ROS, therefore, although injurious in large quantities, can have a paradoxical protective effect within the heart. In this review we provide background information on ROS formation and elimination relevant to anesthetic and adjuvant drugs with particular reference to the heart. The sources of ROS, the means by which they induce cardiac injury or activate protective signaling pathways, the results of clinical studies evaluating ROS scavengers, and the effects of anesthetic drugs on ROS are each discussed. PMID- 16243981 TI - Is the valve OK or not? Immediate evaluation of a replaced aortic valve. AB - Transesophageal echocardiography is a crucial tool in intraoperative evaluation of newly implanted/repaired heart valves because suspected valvular malfunction needs to be identified and sometimes surgically corrected. Although color Doppler is often adequate in evaluating the expected regurgitant jets, as well as excluding pathologic paravalvular leaks, spectral Doppler techniques are the most commonly used methods for estimating transvalvular gradients in the operating room. However, these methods are subject to a variety of confounding factors, including subvalvular gradients and pressure recovery. Other methods of valve area estimation should also be used when evaluating a prostethic aortic valve, including the continuity equation and the left ventricular outflow tract/aortic valve velocity ratio. PMID- 16243982 TI - Transesophageal echocardiographic findings in papillary muscle rupture. PMID- 16243983 TI - An evaluation of bilateral monitoring of cerebral oxygen saturation during pediatric cardiac surgery. AB - Cerebral oximetry is a technique that enables monitoring of regional cerebral oxygenation during cardiac surgery. In this study, we evaluated differences in bi hemispheric measurement of cerebral oxygen saturation using near-infrared spectroscopy in 62 infants undergoing biventricular repair without aortic arch reconstruction. Left and right regional cerebral oxygen saturation index (rSO2i) were recorded continuously after the induction of anesthesia, and data were analyzed at 12 time points. Baseline rSO2i measurements were left 65 +/- 13 and right 66 +/- 13 (P = 0.17). Mean left and right rSO2i measurements were similar (< or =2 percentage points/absolute scale units) before, during, and after cardiopulmonary bypass, irrespective of the use of deep hypothermic circulatory arrest. Further longitudinal neurological outcome studies are required to determine whether uni- or bi-hemispheric monitoring is required in this patient population. PMID- 16243984 TI - The optimal depth of central venous catheter for infants less than 5 kg. AB - To avoid fatal complications of central venous catheterization such as cardiac tamponade, the tip of the central venous catheter (CVC) should be placed outside of the cardiac chamber. To suggest a guideline for a proper depth of CVC in infants, we measured the distance from the skin puncture site to the junction between superior vena cava and right atrium (SVC-RA junction) by using transesophageal echocardiography (TEE). Fifty infants less than 5 kg undergoing surgery for congenital heart disease were enrolled in this prospective study. After the induction of general anesthesia, CVC was inserted via the right subclavian vein. After the tip of the CVC was placed at the SVC-RA junction using TEE guidance, the length of the CVC inserted beneath the skin was measured. The measured distance had a high correlation with the patient's height, weight, and age (r = 0.88, 0.76, and 0.64, respectively). In infants smaller than 5 kg, the following guideline can avoid intraatrial placement of the CVC: a depth between 40 and 45 mm for infants 2.0-3.0 kg in weight, 45-50 mm for those 3.0-3.9 kg, and 50-55 mm for those more than 4.0 kg. PMID- 16243985 TI - Intraarticular bupivacaine-clonidine-morphine versus femoral-sciatic nerve block in pediatric patients undergoing anterior cruciate ligament reconstruction. AB - We hypothesized that combined femoral-sciatic nerve block (FSNB) offers better analgesia with fewer side effects than intraarticular infiltration (IA) in children undergoing anterior cruciate ligament (ACL) reconstruction. Thirty-six children undergoing ACL reconstruction were randomized to FSNB or IA. FSNB patients had FSNB with bupivacaine (0.125%)-clonidine (2 microg/kg), whereas IA patients received bupivacaine (0.25%)-clonidine (1 microg/kg)-morphine (5 mg). Postoperatively, analgesia was provided with patient-controlled analgesia and rescue morphine. Patient demographics were similar. FSNB patients required less intraoperative fentanyl (50 +/- 40 microg versus 80 +/- 50 microg; P = 0.04). Visual analog scale score for FSNB was smaller than IA in the recovery room (1.8 +/- 3 versus 5.4 +/- 3; P = 0.0002) and during the first 24 h (1.6 +/- 1 versus 2.9 +/- 2; P = 0.01)). FSNB morphine use in the first 18 h was less (7 +/- 13 mg versus 21 +/- 21 mg; P = 0.03). Fewer FSNB patients vomited (11% versus 50%; P = 0.03). IA patients required morphine patient-controlled analgesia sooner. After ACL reconstruction in children, FSNB with bupivacaine-clonidine provides better analgesia with fewer side effects than IA with bupivacaine-clonidine-morphine. PMID- 16243986 TI - Large-dose pretreatment with methylprednisolone fails to attenuate neuronal injury after deep hypothermic circulatory arrest in a neonatal piglet model. AB - Conflicting results have been reported with regard to the neuroprotective effects of steroid treatment with cardiopulmonary bypass (CPB) and deep hypothermic circulatory arrest (DHCA). We evaluated the mode and severity of neuronal cell injury in neonatal piglets after prolonged DHCA and the possible neuroprotective effect of systemic pretreatment (>6 h before surgery) with large-dose methylprednisolone (MP). Nineteen neonatal piglets (age, <10 days; weight, 2.1 +/ 0.5 kg) were randomly assigned to 2 groups: 7 animals were pretreated with large dose systemic MP (30 mg/kg) 24 h before surgery, and 12 animals without pharmacological pretreatment (saline) served as control groups. All animals were connected to full-flow CPB with cooling to 15 degrees C and 120 min of DHCA. After rewarming to 38.5 degrees C with CPB, animals were weaned from CPB and survived 6 h before they were killed, and the brain was prepared for light and electron microscopy, immunohistochemistry, and TUNEL-staining. Quantitative histological studies were performed in hippocampus, cortex, cerebellum, and caudate nucleus. Systemic pretreatment with large-dose MP lead to persistent hyperglycemia but no significant changes of cerebral perfusion. Necrotic and apoptotic neuronal cell death were detected in all analyzed brain regions after 120 min of DHCA. In comparison to the control group, large-dose pretreatment with systemic MP lead to an increase of necrotic neuronal cell death and induced significant neuronal apoptosis in the dentate gyrus of the hippocampus (P = 0.001). In conclusion, systemic pretreatment with large-dose MP fails to attenuate neuronal cell injury after prolonged DHCA and induces regional neuronal apoptosis in the dentate gyrus. PMID- 16243987 TI - Total shoulder arthroplasty as an outpatient procedure using ambulatory perineural local anesthetic infusion: a pilot feasibility study. AB - We investigated the feasibility of converting total shoulder arthroplasty (TSA) into an outpatient procedure using ambulatory interscalene perineural ropivacaine infusion. Of the patients of the first phase (n = 8) who were required to remain hospitalized for at least 1 postoperative night, 5 met discharge criteria in the recovery room. Of the subsequent patients of the second phase (n = 6), all met discharge criteria in the recovery room after surgery, and 5 were discharged directly home. For all patients, postoperative pain was well controlled, oral opioid requirements and sleep disturbances were minimal, range-of-motion consistently reached or exceeded the surgeon's expectations, and patient satisfaction was high. These results suggest that TSA may be performed on an outpatient basis using perineural local anesthetic infusion. Additional research is required to define the appropriate subset of patients and assess the incidence of complications associated with this practice before its mainstream use. PMID- 16243988 TI - A randomized, double-blind study of granisetron plus dexamethasone versus ondansetron plus dexamethasone to prevent postoperative nausea and vomiting in patients undergoing abdominal hysterectomy. AB - In this randomized, double-blind study, we evaluated whether small-dose granisetron (0.1 mg) plus dexamethasone 8 mg (G+D) was as effective as ondansetron 4 mg plus dexamethasone 8 mg (O+D) for preventing vomiting during the 0 to 2 h after tracheal extubation in patients undergoing abdominal hysterectomy requiring general anesthesia. Dexamethasone (D) was administered at induction of anesthesia, and granisetron (G) or ondansetron (O) was given approximately 15 min before tracheal extubation. Data on postoperative nausea and vomiting were collected at 0, 2, 6, and 24 h. For the primary efficacy endpoint, most patients in each group had no vomiting in the 0- to 2-h interval (82/87 [94%] for G+D versus 86/89 [97%] for O+D). Effectiveness of G+D was demonstrated versus O+D. Treatment groups were similar with regard to moderate or severe nausea, complete response, rescue medication use, and total control over 24 h. A descriptive assessment of adverse events showed that both combinations were well tolerated with infrequent and similar incidences of adverse events. The combination of small-dose G administered just before tracheal extubation plus D given at induction of anesthesia is an effective alternative to O+D in preventing vomiting during the 0- to 2-h interval after tracheal extubation. PMID- 16243989 TI - Ondansetron, orally disintegrating tablets versus intravenous injection for prevention of intrathecal morphine-induced nausea, vomiting, and pruritus in young males. AB - In this study we compared the efficacy of orally disintegrating tablets (ODT) and IV ondansetron for preventing spinal morphine-induced pruritus and postoperative nausea and vomiting (PONV) in healthy young male patients. Patients who received bupivacaine with 0.20 mg morphine for spinal anesthesia were randomly assigned to the ODT group (ODT ondansetron 8 mg, n = 50), the IV group (4 mg ondansetron IV, n = 50), or the placebo group (n = 50). Each individual was assessed for pruritus, postoperative nausea and vomiting, and pain at 0, 2, 6, 12, 18, and 24 h after surgery using three distinct visual analog scales. The frequencies of postoperative nausea and vomiting and frequencies of requirement for rescue antiemetic and antipruritic were recorded. There were no significant differences among the three groups with respect to incidence or severity of PONV or postoperative pain visual analog scale scores. The incidences of pruritus in the ODT (56%) and IV (66%) groups were significantly different from that in the placebo group (86%) (P < 0.02 for both). Only the ODT group had significantly lower mean pruritus visual analog scale scores at 0, 2, 6, and 12 h postsurgery than the placebo group (P < 0.023 for all). The frequency of requirement for rescue antipruritic was significantly less in the ODT group than the placebo group (P = 0.013). Both ODT ondansetron 8 mg and IV ondansetron 4 mg are more effective than placebo for preventing spinal morphine-induced pruritus, but neither form of this agent reduces spinal morphine-induced postoperative nausea and vomiting in this patient group. PMID- 16243990 TI - The prevalence and significance of low preoperative hemoglobin in ASA 1 or 2 outpatient surgery candidates. AB - Asymptomatic anemia in healthy patients undergoing low risk surgery is rare. In this retrospective study, we examined the records of 9584 ASA class I-II patients scheduled for elective low risk surgery who had a preoperative hemoglobin (hgb) test for the presence of anemia. Hgb <9 g/dL was detected in 75 patients (0.8%). Perioperative management of anemia occurred in no cases of elective surgery in this group. Transfusion of red cells occurred in four other patients, all of whom had hgb >9 g/dL. In all cases, management decisions were based on clinical factors rather than the preoperative hemoglobin test. In healthy patients undergoing low risk elective surgery, routine preoperative hgb testing is not indicated. PMID- 16243991 TI - Opiate-induced nausea and vomiting: what is the challenge? PMID- 16243992 TI - Postoperative nausea and vomiting are strongly influenced by postoperative opioid use in a dose-related manner. AB - We prospectively examined the incidence of postoperative nausea and vomiting (PONV) in a group of 193 elderly surgical inpatients receiving no postoperative antiemetic prophylaxis. Risk factors for PONV and detailed data on postoperative opioid use were recorded. The overall postoperative vomiting (POV) rate was 23.8%, whereas postoperative nausea (PON) was 51.3%. Opioid use (P = 0.025), and female gender (P = 0.038) were identified as significantly influencing POV in this relatively small population. There was a strong logarithmic dose-response relationship between postoperative opioid dose and POV (r2= 0.98, P < 0.01), as well as PON (r2= 0.98, P = 0.01). Use of patient-controlled analgesia or epidural analgesia was a marker for large-dose opioid use (P < 0.001) and was associated with POV in the 24-h postoperative period of 41% and 31% respectively, compared with 11% for other patients (P < 0.001). Future studies defining risk factors for POV should treat postoperative opioid use as a continuous variable, rather than treat it as a dichotomous variable. PMID- 16243994 TI - Rocuronium versus succinylcholine for rapid sequence induction of anesthesia and endotracheal intubation: a prospective, randomized trial in emergent cases. AB - When anesthesia is induced with propofol in elective cases, endotracheal intubation conditions are not different between succinylcholine and rocuronium approximately 60 s after the injection of the neuromuscular relaxant. In the present study, we investigated whether, in emergent cases, endotracheal intubation conditions obtained at the actual moment of intubation under succinylcholine differ from those obtained 60 s after the injection of rocuronium. One-hundred-eighty adult patients requiring rapid sequence induction of anesthesia for emergent surgery received propofol (1.5 mg/kg) and either rocuronium (0.6 mg/kg; endotracheal intubation 60 s after injection) or succinylcholine (1 mg/kg; endotracheal intubation as soon as possible). The time from beginning of the induction until completion of the intubation was shorter after the administration of succinylcholine than after rocuronium (median time 95 s versus 130 s; P < 0.0001). Endotracheal intubation conditions, rated with a 9 point scale, were better after succinylcholine administration than after rocuronium (8.6 +/- 1.1 versus 8.0 +/- 1.5; P < 0.001). There was no significant difference in patients with poor intubation conditions (7 versus 12) or in patients with failed first intubation attempt (4 versus 5) between the groups. We conclude that during rapid sequence induction of anesthesia in emergent cases, succinylcholine allows for a more rapid endotracheal intubation sequence and creates superior intubation conditions compared with rocuronium. PMID- 16243993 TI - Does neostigmine administration produce a clinically important increase in postoperative nausea and vomiting? AB - Neostigmine is used to antagonize neuromuscular blocker-induced residual neuromuscular paralysis. Despite the findings of a previous meta-analysis, the effect of neostigmine on postoperative nausea and vomiting remains unresolved. We reevaluated the effect of neostigmine on postoperative nausea and vomiting while considering the different anticholinergics as potentially confounding factors. We performed a systematic literature search using MEDLINE, Embase, Cochrane library, reference listings, and hand searching with no language restriction through December 2004 and identified 10 clinical, randomized, controlled trials evaluating neostigmine's effect on postoperative nausea and vomiting. Data on nausea or vomiting from 933 patients were extracted for the early (0-6 h), delayed (6-24 h), and overall (0-24 h) postoperative periods and analyzed with RevMan 4.2 (Cochrane Collaboration, Oxford, UK) and multiple logistic regression analysis. The combination of neostigmine with either atropine or glycopyrrolate did not significantly increase the incidence of overall (0-24 h) vomiting (relative risk, 0.91; 95% confidence interval, 0.70-1.18; P = 0.48) or nausea (relative risk, 1.24; 95% confidence interval, 0.98-1.59; P = 0.08). Multiple logistic regression analysis indicated that there was not a significant increase in the risk of vomiting with large compared with small doses of neostigmine. Contrasting a previous analysis, we conclude that there is insufficient evidence to conclude that neostigmine increases the risk of postoperative nausea and vomiting. PMID- 16243995 TI - Nitric oxide is not a mediator of inflammation-induced resistance to atracurium. AB - Resistance to atracurium as a result of increased drug binding to alpha1-acid glycoprotein is associated with increased inducible nitric oxide synthase activity and increased nitric oxide levels in plasma. We investigated if the inhibition of inducible nitric oxide synthase and suppression of nitric oxide can reverse the resistance to atracurium. As a model of alpha1-acid glycoprotein and nitric oxide increase, 84 male Sprague-Dawley rats received an IV injection of either 60 mg/kg Corynebacterium parvum (CP) or saline (control). The 2 groups (CP/Control) were further divided into subgroups, receiving the selective inducible nitric oxide synthase inhibitor, N-Iminolysine, via drinking water at different concentrations. On day 4 post-CP injection, the pharmacodynamics of atracurium were determined. Plasma concentrations of nitric oxide, atracurium, and alpha1-acid glycoprotein were measured and acetylcholine receptor numbers were quantified. In the CP groups, N-Iminolysine suppressed nitric oxide levels in a dose-dependent manner. Resistance to atracurium persisted. alpha1-acid glycoprotein serum levels remained increased in all CP groups with no differences in acetylcholine receptor expression. Our results suggest that the mechanism leading to increased expression of alpha1-acid glycoprotein and consecutive increased protein binding of atracurium is not mediated by inducible nitric oxide synthase induction and nitric oxide expression. PMID- 16243996 TI - Doxapram only slightly reduces the shivering threshold in healthy volunteers. AB - We determined the effects of doxapram on the major autonomic thermoregulatory responses in humans. Nine healthy volunteers were studied on 2 days: control and doxapram (IV infusion to a plasma concentration of 2.4 +/- 0.8, 2.5 +/- 0.9, and 2.6 +/- 1.1 microg/mL at the sweating, vasoconstriction, and shivering thresholds, respectively). Each day, skin and core temperatures were increased to provoke sweating, then reduced to elicit peripheral vasoconstriction and shivering. We determined the sweating, vasoconstriction, and shivering thresholds with compensation for changes in skin temperature. Data were analyzed with paired t-tests and presented as mean +/- sd; P < 0.05 was considered statistically significant. Doxapram did not change the sweating (control: 37.5 degrees +/- 0.4 degrees C, doxapram: 37.3 degrees +/- 0.4 degrees C; P = 0.290) or the vasoconstriction threshold (36.8 degrees +/- 0.7 degrees C versus 36.4 degrees +/ 0.5 degrees C; P = 0.110). However, it significantly reduced the shivering threshold from 36.2 degrees +/- 0.5 degrees C to 35.7 degrees +/- 0.7 degrees C (P = 0.012). No sedation or symptoms of panic were observed on either study day. The observed reduction in the shivering threshold explains the drug's efficacy for treatment of postoperative shivering; however, a reduction of only 0.5 degrees C is unlikely to markedly facilitate induction of therapeutic hypothermia as a sole drug. PMID- 16243997 TI - Clonidine premedication in patients with sleep apnea syndrome: a randomized, double-blind, placebo-controlled study. AB - Patients with sleep apnea often present with cardiac diseases and breathing difficulties, with a high risk of postoperative respiratory depression. We conducted a randomized, double-blind, prospective study in 30 adult patients with obstructive sleep apnea, undergoing elective ear-nose-throat surgery. The patients were randomly assigned to receive placebo or clonidine (2 microg/kg oral) the night before and the next morning 2 h before surgery. Spo2, heart rate, mean arterial blood pressure, snoring, and oronasal airflow were monitored for 36 h. A standard anesthesia was used consisting of propofol and remifentanil. Anesthetic drug consumption, postoperative analgesics, and pain score were recorded. In the clonidine group, mean arterial blood pressures were significantly lower during induction, operation, and emergence from anesthesia. Both propofol dose required for induction (190 +/- 32.2 mg) and anesthesia (6.3 +/- 1.3 mg . kg(-1).h(-1)) during surgery were significantly reduced in the clonidine group compared with the placebo group (induction 218 +/- 32.4, anesthesia 7.70 +/- 1.5; P < 0.05). Piritramide consumption (7.4 +/- 5.1 versus 14.2 +/- 8.5 mg; P < 0.05) and analgesia scores were significantly reduced in the clonidine group. Apnea and desaturation index were not different between the groups, whereas the minimal postoperative oxygen saturation on the day of surgery was significantly lower in the placebo than in the clonidine group (76.7% +/- 8.0% versus 82.4% +/- 5.8%; P < 0.05). We conclude that oral clonidine premedication stabilizes hemodynamic variables during induction, maintenance, and emergence from anesthesia and reduces the amount of intraoperative anesthetics and postoperative opioids without deterioration of ventilation. PMID- 16243998 TI - The effects of anesthetics and ethanol on alpha2 adrenoceptor subtypes expressed with G protein-coupled inwardly rectifying potassium channels in Xenopus oocytes. AB - A wide range of physiological effects are mediated by alpha2-adrenoceptors (ARs) through their association with G protein-coupled inwardly rectifying potassium (GIRK) channels. Although alpha2-ARs are divided into three subtypes (alpha2A-C), a pharmacological distinction among the subtypes is difficult to establish because of the lack of a selective agonist and antagonist; therefore, little is known about the effects of anesthetics on the alpha2-AR subtypes. We expressed each subtype together with GIRK1/GIRK2 subunits in Xenopus oocytes and observed alpha2-AR-mediated GIRK1/GIRK2 currents to test the effects of ethanol, halothane, and several IV anesthetics at clinical concentrations. UK 14,304, a selective alpha2-AR agonist, evoked GIRK1/GIRK2 currents in every subtype. None of the IV anesthetics, which included pentobarbital, propofol, ketamine, and alphaxalone, influenced UK 14,304-evoked potassium currents in any of the receptor subtypes. Ethanol enhanced the UK 14,304-evoked potassium currents, whereas halothane inhibited the currents. However, these effects were not significantly different from those on the baseline-GIRK1/GIRK2 current, suggesting that neither ethanol nor halothane acts directly on the alpha2-AR subtypes. Although none of the drugs examined had any effect on the alpha2-ARs, the physiological actions of the alpha2-ARs mediated by the GIRK1/GIRK2 channels may be affected by ethanol and halothane. PMID- 16243999 TI - Spatial memory performance 2 weeks after general anesthesia in adult rats. AB - We have previously demonstrated that general anesthesia with 1.2% isoflurane-70% nitrous oxide impairs acquisition of a radial arm maze task in both young and aged rats when testing begins 2 days after anesthesia and in aged rats when testing begins 2 wk later. We designed this study to examine whether postanesthesia learning impairment is persistent in young rats. Six-month-old rats were randomized to anesthesia for 2 h with 1.2% isoflurane-70% nitrous oxide, 1.8% isoflurane, or a control group that received 30% oxygen (n = 10 per group). Rats recovered for 2 wk and were then tested daily on a radial arm maze for 14 days. There were no differences between the controls and anesthesia groups in number of correct choices to first error or time to complete the maze. There was no main effect of group in terms of total number of errors (P > 0.05) but the group by day interaction was significant (P < 0.05), reflecting improved performance in the 1.2% isoflurane-70% nitrous oxide group relative to controls during the later days of testing (P < 0.005). Hence, in adult rats, previous general anesthesia is not associated with impaired learning 2 wk later. In fact, previous 1.2% isoflurane-70% nitrous oxide improves maze performance 2 wk later. PMID- 16244000 TI - Modern wireless telecommunication technologies and their electromagnetic compatibility with life-supporting equipment. AB - Hospitals rely on pagers and ordinary telephones to reach staff members in emergency situations. New telecommunication technologies such as General Packet Radio Service (GPRS), the third generation mobile phone system Universal Mobile Telecommunications System (UMTS), and Wireless Local Area Network (WLAN) might be able to replace hospital pagers if they are electromagnetically compatible with medical devices. In this study, we sought to determine if GPRS, UMTS (Wideband Code Division Multiple Access-Frequency Division Duplex [WCDMA FDD]), and WLAN (IEEE 802.11b) transmitted signals interfere with life-supporting equipment in the intensive care and operating room environment. According to United States standard, ANSI C63.18-1997, laboratory tests were performed on 76 medical devices. In addition, clinical tests during 11 operations and 100 h of intensive care were performed. UMTS and WLAN signals caused little interference. Devices using these technologies can be used safely in critical care areas and during operations, but direct contact between medical devices and wireless communication devices ought to be avoided. In the case of GPRS, at a distance of 50 cm, it caused an older infusion pump to alarm and stop infusing; the pump had to be reset. Also, 10 cases of interference with device displays occurred. GPRS can be used safely at a distance of 1 m. Terminals/cellular phones using these technologies should be allowed without restriction in public areas because the risk of interference is minimal. PMID- 16244001 TI - Denaturing high performance liquid chromatography screening of ryanodine receptor type 1 gene in patients with malignant hyperthermia in Taiwan and identification of a novel mutation (Y522C). AB - We performed the present study to identify the mutation in patients in Taiwan with malignant hyperthermia (MH). We also test the hypothesis that a denaturing high-performance liquid chromatography (DHPLC) protocol can be used for mutation detection in these patients. We identified five Taiwanese patients with typical clinical presentations of MH after general anesthesia. We also enrolled 50 healthy volunteers. Polymerase chain reaction was used to amplify the ryanodine receptor (RYR1) gene mutation hot spots and DHPLC techniques were used to screen for mutations. Upon detection of a heterozygous elution pattern in DHPLC analysis, DNA sequencing reaction was performed to identify the nucleotide variations. We identified a RYR1 mutation in all 5 patients with MH. There were 4 different mutations in the 5 patients: Tyr522Cys, Arg552Trp, Val2168Met, and Thr2206Arg. Among the 5 patients, 2 unrelated patients had the same Thr2206Arg mutation. Three of the mutations had been reported before, but the Tyr522Cys mutation was novel. None of the MH-related mutations were found in the control group. In conclusion, we identified RYR1 mutations in 5 Taiwanese patients with MH using a DHPLC-based approach. A DHPLC-based genetic test may be developed as a noninvasive and convenient test for MH. PMID- 16244002 TI - Reducing the incidence of surgical fires: supplying nasal cannulae with sub-100% O2 gas mixtures from anesthesia machines. AB - In June 2003, the Joint Commission on Accreditation of Healthcare Organizations (JCAHO) recommended: "As a general policy, use air or FiO2 at < or =30% for open delivery (consistent with patient needs)" to prevent surgical fires. One way to interpret JCAHO's recommendation is that 100% O2 should not be indiscriminately used, and anesthesia providers should have the ability, consistent with patient needs and their clinical judgment, to deliver sub-100% O2 with nasal cannulae. An auxiliary O2 flowmeter has a barbed outlet connector that offers a convenient means to connect a cannula to an anesthesia machine and is routinely used for open delivery of 100% O2. The auxiliary O2 flowmeter provides only 100% O2 and thus does not allow titration of the O2 concentration to patient needs and may increase the risk of surgical fires. This report clarifies the JCAHO recommendation and describes different means of addressing it that are based primarily on using the anesthesia machine to blend a sub-100% O2 gas mixture and delivering it via a nasal cannula. The options presented depend on the model and manufacturer of the anesthesia machine and allow delivery via nasal cannula of O2 concentrations that range from 21% to 100%. PMID- 16244003 TI - Air detection performance of the level 1 H-1200 fluid and blood warmer. AB - We evaluated the Level 1 H-1200 fluid warmer during simulated conditions of minor to massive air embolism. The fluids we tested were crystalloid and diluted red cells (estimated hematocrit 50%) during gravity and pressure driven flow. The air volumes tested ranged from 1-60 mL for crystalloid and 30-150 mL for red cells. No air was observed distal to the air detector and clamp during all test conditions. The device consistently alarmed and automatically shut off flow. Air was easily purged through the gas vent-filter assembly during simulated air embolism with crystalloid but not with red cell infusion. The use of ultrasonic air detection coupled with automatic shutoff is a significant safety improvement of the Level 1 H-1200 fluid and blood warmer. PMID- 16244004 TI - Antinociception of intrathecal adenosine receptor subtype agonists in rat formalin test. AB - Adenosine has shown antinociceptive action via spinal adenosine receptors. There are four types of adenosine receptors: A1, A2A, A2B, and A3. We characterized the nature of types of adenosine receptors for the control of nociception at the spinal level. For nociception, formalin solution (5%, 50 microL) was injected into the hindpaw of male Sprague-Dawley rats. The effects of intrathecal adenosine A1 (CPA), A2A (DPMA), and A3 (IB-MECA) receptor agonists were examined. CPA and IB-MECA produced limited or no effect on the early phase response of the formalin test, respectively, but the two drugs depressed the late phase response. DPMA suppressed both phase responses. CPA was the most potent drug among the three in the late phase. These results suggest that spinal adenosine A1 and A2A receptors may be involved in the modulation of the early and the late phase responses of the formalin test, whereas adenosine A3 receptor may be involved in the regulation of the late phase response. PMID- 16244005 TI - The neurological safety of epidural gabapentin in rats: a light microscopic examination. AB - Gabapentin acts primarily on the central nervous system. Therefore, we hypothesized that the direct epidural administration of gabapentin could have various advantages over its oral administration with respect to required dose, side effects, and efficacy. However, before administering gabapentin into the epidural space in a clinical setting, its neurotoxicity must be examined in animals. Thus, we evaluated neurotoxicity of epidural gabapentin by observing behavioral and sensory-motor changes, and by histopathological examinations of spinal cords and dorsal root ganglia in the rat. Twenty-seven rats were randomly divided into 3 groups, which were administered 0.3 mL (30 mg) of epidural gabapentin (group G, n = 9) and the same volume of epidural alcohol (group A, n = 9) or normal saline (group N, n = 9). No rats in groups G and N showed sensory motor dysfunction, behavioral change, or histopathological abnormalities over a 3 wk observation period, whereas all rats in group A showed abnormalities. We conclude that the direct epidural injection of gabapentin in rats did not show any neurotoxic evidence in terms of sensory-motor functions and behavior, or by a microscopic histopathological evaluation. This study represents a first promising step toward the trial of epidural gabapentin in a clinical setting. PMID- 16244006 TI - Multimodal analgesia with gabapentin and local anesthetics prevents acute and chronic pain after breast surgery for cancer. AB - We evaluated the effect of multimodal analgesia on acute and chronic pain after breast surgery for cancer. Fifty patients scheduled for breast cancer surgery were blindly randomized to receive gabapentin, eutectic mixture of local anesthetics cream, and ropivacaine in the wound or three placebos. Pain (visual analog scale) and analgesics were recorded in the postanesthesia care unit (PACU) 3, 6, and 9 h and 8 days after surgery. Three and 6 mo later, patients were assessed for chronic pain. The treatment group consumed less paracetamol in the PACU (469 versus 991 mg; P < 0.002) and less Lonalgal (1.0 versus 4.4 tablets; P = 0.003) than the controls, exhibited lower visual analog scale scores at rest in the PACU (P = 0.001) and on postoperative Days 1, 3, and 5 (P = 0.040, P = 0.015, and P = 0.045, respectively), and after movement in the PACU (P = 0.001) and on postoperative Days 2, 4, and 8 (P = 0.028, P = 0.007, and P = 0.032, respectively). Three and 6 mo after surgery, 18 of 22 (82%) and 12 of 21 (57%) of the controls reported chronic pain versus 10 of 22 (45%) and 6 of 20 (30%) in the treatment group (P = 0.028 and P = 0.424, respectively); 5 of 22 and 4 of 21 of the controls required analgesics versus 0 of 22 and 0 of 20 of those treated (P = 0.048 and P = 0.107, respectively). Multimodal analgesia reduced acute and chronic pain after breast surgery for cancer. PMID- 16244008 TI - Sacroiliac joint pain: a comprehensive review of anatomy, diagnosis, and treatment. AB - Sacroiliac (SI) joint pain is a challenging condition affecting 15% to 25% of patients with axial low back pain, for which there is no standard long-term treatment. Recent studies have demonstrated that historical and physical examination findings and radiological imaging are insufficient to diagnose SI joint pain. The most commonly used method to diagnose the SI joint as a pain generator is with small-volume local anesthetic blocks, although the validity of this practice remains unproven. In the present review I provide a comprehensive review of the anatomy, function, and mechanisms of injury of the SI joint, along with a systematic assessment of its diagnosis and treatment. PMID- 16244007 TI - The effects of intraarticular resiniferatoxin in experimental knee-joint arthritis. AB - In this study we sought to determine whether an intraarticular administration of a vanilloid agonist resiniferatoxin (RTX) produces an analgesic effect in experimental arthritis. Knee joint inflammation was induced in rats by intraarticular carrageenan (2%, 30 microL). Pain score and left/right hind leg weight distribution ratio were used to assess pain behavior. Changes in knee dimensions were evaluated by measuring external circumference and intraarticular area (ultrasound scanning). The intraarticular administration of RTX (0.0003% or 0.003%, 30 microL) provided a significant analgesic effect. Twenty-four hours after RTX administration, the pain score was reduced from 15.1 +/- 4.7 to 6.9 +/- 4.4 (P < 0.01) with 0.0003% and was abolished (P < 0.0001) with 0.003%. The improvement in weight distribution ratio lasted for several days after the RTX administration. Reduction in knee circumference demonstrated that intraarticular RTX suppressed the carrageenan-induced edema by at least one third. Ultrasound scanning revealed no RTX-induced decrease of the intraarticular area. The experiments demonstrated that intraarticular RTX inhibits pain behavior in knee joint arthritis and that this effect is dose-dependent. These results suggest a new direction for peripheral analgesia. PMID- 16244009 TI - The legend of the P value. PMID- 16244010 TI - Chemical dependency treatment outcomes of residents in anesthesiology: results of a survey. AB - Substance abuse is a potentially lethal occupational hazard confronting anesthesiology residents. We present the results of a survey sent to all United States anesthesiology training programs regarding experience with and outcomes of chemically dependent residents from 1991 to 2001. The response rate was 66%. Eighty percent reported experience with impaired residents and 19% reported at least one pretreatment fatality. Despite this familiarity, few programs required pre-employment drug testing or used substance abuse screening tools during interviews. The majority of impaired residents attempted reentry into anesthesiology after treatment. Only 46% of these were successful in completion of anesthesiology residency. Eventually, 40% of residents who underwent treatment and returned to medical training entered another specialty. The mortality rate for the remaining anesthesiology residents was 9%. Long-term outcome was reported for 93% of all treated residents. Of these, 56% were successful in some specialty of medicine at the end of the survey period. We hypothesize that specialty change afforded substantial improvement in the overall success rate and avoided significant mortality. Redirection of rehabilitated residents into lower-risk specialties may allow a larger number to achieve successful medical careers. PMID- 16244011 TI - Brain and sepsis: functional impairment, structural damage, and markers. PMID- 16244012 TI - Slight increase of serum S-100B during porcine endotoxemic shock may indicate blood-brain barrier damage. AB - Septic shock is a condition that affects many organs, but little is known about the effects on the central nervous system. S-100B, an acidic low molecular weight protein, has attracted considerable interest as a marker for brain damage and disintegration of the blood-brain barrier. It is released into the cerebrospinal fluid and blood from brain tissue after brain damage. We studied S-100B in a porcine model of endotoxemic shock that resembles human Gram-negative septic shock. Ten piglets received IV endotoxin, and plasma samples were collected before the endotoxin infusion and each hour (1-6 h) during the endotoxin infusion. S-100B was measured by sandwich enzyme-linked immunosorbent assay. Low levels of plasma S-100B were detected, but there was a significant increase in S 100B during Hours 1-5 in comparison with the 0 values. We determined that endotoxemia causes a very small but significant increase in the levels of the widely used brain damage marker serum S-100B. However, it cannot be excluded that the increase in S-100B could be caused by release from organs other than the brain. PMID- 16244013 TI - Validation of a behavioral pain scale in critically ill, sedated, and mechanically ventilated patients. AB - Assessing pain in critically ill patients, particularly in nonverbal patients, is a great challenge. In this study, we validated a behavioral pain scale (BPS) in critically ill, sedated, and mechanically ventilated patients. The BPS score was the sum of 3 subscales that have a range score of 1-4: facial expression, upper limb movements, and compliance with mechanical ventilation. Two assessors observed and scored pain simultaneously with the BPS at rest and during painful procedures. The psychometric properties of the BPS that were studied were reliability, validity, and responsiveness. We achieved 360 observations in 30 patients. The BPS was internally reliable (Cronbach alpha = 0.72). The intraclass correlation coefficient to evaluate inter-rater reliability was high (0.95). Validity was demonstrated by the change in BPS scores, which were significantly higher during painful procedures, with averages of 3.9 +/- 1.1 at rest and 6.8 +/ 1.9 during procedures (P < 0.001), and by the principal components factor analysis, which revealed a large first-factor accounting for 65% of the variance in pain expression. The BPS exhibited excellent responsiveness, with an effect size ranging from 2.2 to 3.4. This study demonstrated that the BPS can be valid and reliable for measuring pain in noncommunicative intensive care unit patients. PMID- 16244014 TI - A comparison of the laryngeal tube and bag-valve mask ventilation by emergency medical technicians: a feasibility study in anesthetized patients. AB - Airway management is of major importance in emergency care. The basic technique for all health care providers is bag-valve mask (BVM) ventilation, which requires skill and may be difficult to perform. Endotracheal intubation, which is the advanced method for securing the airway, is a demanding technique that has been shown to be associated with infrequent success, even when used by experienced paramedical personnel. Therefore, alternative airway devices have been sought. The use of the laryngeal tube (LT) by experienced anesthesia personnel had been studied in anesthetized patients and manikins in emergency medical training. We decided to evaluate the ability of inexperienced firefighter-emergency medical technician students (fire-EMT) to insert the LT or perform BVM in anesthetized patients. Thirty fire-EMTs randomly inserted the LT (n = 15) and performed 1 min of ventilation or used the BVM (n = 15). We found that all students successfully (100%) inserted the LT. Those who inserted the LT on the first attempt (73%) required 48.2 +/- 14.7 s for the insertion. Both the LT and BVM provided adequate oxygenation and ventilation. In this study, we found that inexperienced fire-EMT students inserted LT and performed 1-min ventilation with a reasonable success rate and insertion time in anesthetized patients. PMID- 16244015 TI - A rat model for isolated bilateral lung contusion from blunt chest trauma. AB - Lung contusion affects 17%-25% of adult blunt trauma patients, and is the leading cause of death from blunt thoracic injury. A small animal model for isolated bilateral lung contusion has not been developed. We induced lung contusion in anesthetized rats by dropping a 0.3-kg weight onto a precordial protective shield to direct the impact force away from the heart and toward the lungs. Lung injury was characterized as a function of chest impact energy (1.8-2.7 J) by measurements of arterial oxygenation, bronchoalveolar lavage (BAL) albumin and cytology, pressure-volume mechanics, and histopathology. Histology confirmed bilateral lung contusion without substantial cardiac muscle trauma. Rats receiving 2.7 J of chest impact energy had 33% mortality that exceeded prospectively defined limits for sublethal injury. Hypoxemia in rats with maximal sublethal injury (2.45 J) met criteria for acute lung injury at < or =24 h, improving by 48 h. BAL albumin levels were highest at < or =24 h, and remained elevated along with increased BAL leukocytes and decreased lung volumes at 48 h. We concluded that an impact energy of 2.45 J induces isolated, bilateral lung contusion and provides a useful model for future mechanistic pathophysiological assessments. PMID- 16244016 TI - Delayed respiratory depression after risperidone overdose. AB - Risperidone is an atypical antipsychotic drug used for the treatment of schizophrenia. Both positive and negative symptoms are prominent with its use. Metabolism occurs mainly in the liver, where risperidone is changed by CUP2D6 to an active metabolite, 9-hydroxyrisperidone. The half-lives of risperidone and its metabolite are 3 and 7 h, respectively. Genetic polymorphism is seen in the 6%-8% of white patients who are considered poor metabolizers. In poor metabolizers, the half-life extends to 20-30 h. We present an unusual case of unanticipated delayed respiratory depression after risperidone overdose. PMID- 16244017 TI - The efficacy of postoperative ondansetron (Zofran) orally disintegrating tablets for preventing nausea and vomiting after acoustic neuroma surgery. AB - Postoperative nausea and vomiting is a frequent complication of craniotomy. We evaluated the ability of intraoperative IV ondansetron followed by postoperative ondansetron in an orally disintegrating tablet formulation to reduce the frequency and severity of postoperative nausea and vomiting in a prospective, randomized, placebo-controlled double-blind trial of 60 patients undergoing acoustic neuroma resection. Each patient received intraoperative ondansetron (4 mg IV) or placebo 30 min before case end. Postoperatively, patients received ondansetron in an orally disintegrating tablet formulation (8 mg BID) or placebo twice a day for up to 72 h. Metoclopramide was available as rescue therapy for both groups. Severity of nausea (as measured on a 10-cm visual scale), number of emetic episodes, and requirement for rescue therapy were recorded. In the immediate postoperative period, nausea severity was less in patients treated with ondansetron than placebo (3.3 +/- 4.1 versus 7.3 +/- 4.2; P < 0.001) and fewer patients experienced vomiting (3 of 28 versus 11 of 32; chi2 P < 0.01). More patients required some form of rescue treatment in the placebo group on the first postoperative day (26 of 32 versus 16 of 28; chi2 P < 0.01). We conclude that after acoustic neuroma surgery IV ondansetron treatment prevents immediate postoperative nausea and vomiting. Postoperative treatment with ondansetron in an orally disintegrating tablet formulation was associated with less frequent rescue therapy as compared with placebo on the first postoperative day. PMID- 16244018 TI - Epidural blood patch for headache after lumboperitoneal shunt placement. AB - Headaches complicating lumboperitoneal (LP) shunt placement have been attributed to shunt failure with resultant high intracranial pressure or to overdrainage with resultant low intracranial pressure. In this case, a 17-yr-old girl had symptoms of a low-pressure headache after LP shunt placement alleviated by an epidural blood patch. The success of this therapy suggests postdural puncture as a possible cause for low-pressure headache after LP shunt placement. Epidural blood patch may be an alternative initial therapy for some low-pressure headaches after LP shunt placement. PMID- 16244019 TI - Postural headache in the presence of cerebral venous sinus thrombosis. AB - Cerebral venous sinus thrombosis (CVST) can present with a headache similar to that after a dural puncture. We report on a patient who developed postural headache after epidural anesthesia for delivery. The headache became more intense during the following 6 days, and the patient had a tonic clonic seizure. A magnetic resonance angiogram demonstrated CSVT, and anticoagulation therapy was started, with resolution of the symptoms over 2 wk. Any postdural-puncture headache that loses its positional character, becomes persistent, or does not improve with a properly performed blood patch should raise the suspicion of CVST. PMID- 16244021 TI - The efficacy of plethysmographic pulse wave amplitude as an indicator for intravascular injection of epinephrine-containing epidural test dose in anesthetized adults. AB - In this study, I evaluated the efficacy of plethysmographic pulse wave amplitude (PPWA) in detecting intravascular injection of a simulated epidural test dose containing 15 microg of epinephrine in adults during either sevoflurane or isoflurane inhaled anesthesia and compared its reliability to the classical heart rate (HR; positive if > or =10 bpm) and systolic blood pressure (SBP; positive if > or =15 mm Hg) criteria. Eighty patients were randomized to receive either 1 mean alveolar anesthetic concentration of sevoflurane or 1 mean alveolar anesthetic concentration of isoflurane (n = 40 for each anesthesia group). Patients in each anesthesia group microg of epinephrine IV or 3 mL of saline IV (n = 20 each). HR, SBP, and PPWA were monitored for 5 min after injection. Injection of the test dose resulted in peak PPWA decrease by 61% +/- 17% and 58% +/- 15% at 61 +/- 12 s and 63 +/- 13 s in the sevoflurane and isoflurane groups, respectively. Positive PPWA criterion, as determined from peak increases during saline administration, was a decrease in PPWA > or =10%. Using this value, the sensitivity, specificity, positive predictive, and negative predictive values of PPWA were 100% in both anesthetic groups. On the contrary, sensitivities of 85% and 95% were obtained based on HR criterion in the sevoflurane and isoflurane patients, respectively, and a sensitivity of 90% was obtained in both anesthesia groups on the basis of SBP criterion. In conclusion, PPWA is a reliable alternative to conventional hemodynamic criteria for detection of an intravascular injection of epidural test dose. PMID- 16244020 TI - Administration of local anesthetic through the epidural needle before catheter insertion improves the quality of anesthesia and reduces catheter-related complications. AB - Epidural catheter placement offers flexibility in block management. However, during epidural catheter insertion, complications such as paresthesia and venous and subarachnoid cannulation may occur, and suboptimal catheter placement can affect the quality of anesthesia. We performed this prospective, randomized, double-blind study to assess the effect of a single-injection dose of local anesthetic (20 mL of 2% lidocaine) through the epidural needle as a priming solution into the epidural space before catheter insertion. We randomized 240 patients into 2 equal groups and measured the quality of anesthesia and the incidence of complications. In the needle group (n = 100), catheters were inserted after injection of a full dose of local anesthetic through the needle. In the catheter group (n = 98), the catheters were inserted immediately after identification of the epidural space. Local anesthetic was then injected via the catheter. We noted the occurrence of paresthesia, inability to advance the catheter, or IV or subarachnoid catheter placement. Sensory and motor block were assessed 20 min after the injection of local anesthetic. Surgery was initiated when adequate sensory loss was confirmed. In the catheter group, the incidence of paresthesia during catheter placement was 31.6% compared with 11% in the needle group (P = 0.00038). IV catheterization occurred in 8.2% versus 2% of patients in the catheter and needle groups, respectively (P = 0.048). More patients in the needle group had excellent surgical conditions than the catheter group (89.6% versus 72.9; P < 0.003). We conclude that giving a single-injection dose via the epidural needle before catheter placement improves the quality of epidural anesthesia and reduces catheter-related complications. PMID- 16244022 TI - Small-dose bupivacaine-sufentanil prevents cardiac output modifications after spinal anesthesia. AB - Spinal injection of small-dose (SD) bupivacaine decreases the likelihood of hypotension compared with large-dose (LD) bupivacaine. We assumed that a SD of bupivacaine could also prevent the decrease in cardiac output (CO). Patients undergoing elective urologic, lower abdominal, or lower limb surgery under spinal anesthesia were included in this prospective randomized study. Spinal injection consisted of 5 mug of sufentanil and either SD (7.5 mg of hyperbaric bupivacaine with glucosemonohydrate 80 mg/mL; n = 19 patients) or LD (12.5 mg of hyperbaric bupivacaine with glucosemonohydrate 80 mg/mL; n = 19 patients). CO (impedance cardiography), arterial blood pressure, and heart rate) were measured at 1 min before performance of spinal block and 2, 10, and 30 min after the intrathecal injection. Sensory level was also assessed at 30 min. CO was higher in the SD group as compared with the LD group from 2 min to 30 min after spinal anesthesia. Moreover, CO increased at 2 min in the SD group and decreased at 10 and 30 min in the LD group compared with baseline value. In conclusion, SD bupivacaine provides successful anesthesia and gives better CO stability than LD. PMID- 16244023 TI - Prophylactic intravenous ondansetron and dolasetron in intrathecal morphine induced pruritus: a randomized, double-blinded, placebo-controlled study. AB - Pruritus is the most common side effect of intrathecal morphine for postoperative pain relief. Activation of central 5-hydroxytryptamine subtype 3 (5-HT3) receptors is one of its possible mechanisms. The role of 5-HT3 antagonists in the prevention of pruritus has not been clearly established. In a prospective, randomized, double-blind, placebo-controlled study, we evaluated the efficacy of prophylactic administration of ondansetron and dolasetron for the prevention of intrathecal morphine-induced pruritus. The patients were randomized into 3 groups to receive either 4 mg ondansetron IV (group O, n = 35), 12.5 mg dolasetron IV (group D, n = 35) or 5 mL placebo (group P, n = 35) 30 min before administration of spinal anesthesia with 10 to 17.5 mg of 0.5% hyperbaric bupivacaine and 0.25 mg of morphine for urologic, orthopedic, or vascular surgery. Patients were evaluated for incidence and severity of pruritus at arrival to the postanesthesia care unit and at 2, 4, 8, and 24 h postoperatively. The incidence and severity of pruritus was significantly less frequent in the ondansetron and dolasetron groups compared with placebo (34%, 20%, and 66% respectively, P < 0.01). Patients who received 5-HT3 antagonist reported significantly less total severity of pruritus compared with placebo during the first 8 h and the severe pruritus was observed only in patients within P group (P group: 4 of 35; 11%, O or D group: 0 of 35; 0%, P < 0.05). We conclude that the prophylactic use of ondansetron and dolasetron helps to reduce the incidence and severity of intrathecal morphine induced pruritus. PMID- 16244024 TI - The effects of continuous epidural anesthesia and analgesia on stress response and immune function in patients undergoing radical esophagectomy. AB - We investigated whether perioperative extensive epidural block (C3-L) affects postoperative immune response in patients undergoing radical esophagectomy. Patients undergoing radical esophagectomy were randomly assigned to either general anesthesia with continuous epidural infusion via 2 epidural catheters that was continued for postoperative analgesia (group E, n = 15) or intraoperative general anesthesia and postoperative IV morphine analgesia (group G, n = 15). Plasma levels of stress hormones, cytokines, C-reactive protein (CRP), leukocyte counts, and distribution of lymphocyte subsets were assessed before and after surgery and on postoperative days (PODs) 1 and 3. In comparison with group E, significant increases in plasma epinephrine level at the end of surgery (P < 0.05) and norepinephrine level at the end of surgery (P < 0.01) and on POD1 (P < 0.01) and POD3 (P < 0.01) and significant decrease in cluster of differentiation (CD4/CD8 ratio) at the end of surgery (P < 0.05) were observed in group G. However, there were no significant differences in other variables between groups. In both groups, plasma cortisol, adrenocorticotropic hormone, interleukin (IL)-1beta, IL-6, IL-10, and CRP levels were increased after surgery (each group P < 0.01) and IL-1beta, IL-6, IL-10, and CRP were still increased on POD1 and POD3 (each change, each group P < 0.01). Leukocyte counts were increased on POD1 (each group P < 0.05) and POD3 (each group P < 0.01). The proportion of lymphocytes decreased from the end of surgery to POD3 (each group P < 0.01). The proportion of B cells was increased on POD1 (each group P < 0.01); that of natural killer cells was decreased at POD1 and POD3 (each group P < 0.01). We conclude that tissue damage and inflammation apparently overcome the effects of extensive epidural block on stress response and immune function in radical esophagectomy. PMID- 16244025 TI - An in vitro comparison of the electrical conducting properties of multiport versus single-port epidural catheters for the epidural stimulation test. AB - Effective conduction of electricity through a catheter is essential for the success of the epidural stimulation test. In this in vitro study we examined the electrical conductivity of single and multiport epidural catheters (with and without embedded metal elements) after being primed with normal saline. Seven different types of 19-gauge catheters (n = 5), either single-port or multiport catheters, with or without embedded metal elements, were studied. The proximal end of each epidural catheter was connected to the cathode of a nerve stimulator via an electrode adapter. The catheter, primed with normal saline, was placed at the bottom of a syringe filled with 5 different volumes of saline (1, 2, 3, 4 and 5 mL) and attached to an electrode adapter. The voltage of the peripheral nerve stimulator was measured using an oscilloscope. The electrical resistance between the proximal and distal end of the catheter was calculated using Ohm's Law. In catheters without metal elements the electrical resistances were too high to be measured. In catheters that had metal elements, the mean electrical resistances of the same catheter design (single-port or multiport) were similar. However, the electrical resistances of the multiport metal reinforced epidural catheters were significantly lower (P < 0.05) than the single-port metal coil reinforced epidural catheters. The volume of saline in the syringe had no impact on the measured electrical resistances. This study suggests that multiport metal reinforced epidural catheters have low electrical resistances and, thus, are a reasonable alternative to single-port catheters for transmitting sufficient current for performing the epidural stimulation test. On the other hand, epidural catheters without metal elements (single-port or multiport) are not suitable for performing the stimulation test. PMID- 16244026 TI - Continuous maxillary and mandibular nerve block for perioperative pain relief: the excision of a complicated pleomorphic adenoma. AB - A 68-yr-old hypertensive patient with ischemic heart disease and intractable atrial fibrillation with stable hemodynamics and poor chest compliance underwent pleomorphic adenoma of the parotid gland using regional anesthesia with continuous propofol sedation. Continuous maxillary and mandibular nerve blocks were performed and excision was performed with complete intraoperative and postoperative pain relief without compromising the patient's hemodynamic variables. PMID- 16244027 TI - The feasibility of laryngoscope-guided tracheal intubation in microgravity during parabolic flight: a comparison of two techniques. AB - We determined the feasibility of laryngoscope-guided tracheal intubation (LG-TI) in microgravity obtained during parabolic flight and tested the hypothesis that LG-TI is similarly successful in the free-floating condition, with the patient's head gripped between the anesthesiologist's knees, as in the restrained condition, with the torso strapped to the surface. Three personnel with no experience in airway management or microgravity participated in the study. LG-TI of a sophisticated full-size manikin was attempted on seven occasions in each condition by each investigator after ground-based training. The parabolic flights, which took place in an Airbus 300 over the Atlantic Ocean, provided 23 s of microgravity. During this time, the investigator opened a box with airway equipment, performed LG-TI, and attached and held onto a self-inflating bag. The efficacy of ventilation was assessed during level flight by squeezing the bag and noting whether the manikin sensors indicated a tidal volume > or =300 mL. There were no differences in ventilation success (41% versus 33%) or time to successful insertion (both 18 s) between the free-floating and the restrained conditions. More than 90% of failures were caused by the inability to insert the tracheal tube within 23 s. There were no differences in performance among investigators. We conclude that LG-TI is feasible in microgravity obtained during parabolic flight, but the success rate is infrequent because of severe time restrictions. There were no differences in success rate between the free-floating condition, with the head gripped between the knees, and the restrained condition, with the torso strapped to the surface. PMID- 16244028 TI - Alkalinization of intracuff lidocaine: efficacy and safety. AB - When alkalinized lidocaine instead of air is used to fill the endotracheal tube (ETT) cuff, coughing, and bucking are decreased during extubation when ventilation is controlled with N2O. However, sodium bicarbonate (NaHCO3) used to transform lidocaine hydrochloride (L-HCl) to lidocaine base induces a pH increase that could be irritating for mucosa in the case of cuff rupture. Therefore, we determined, in a randomized controlled study with controlled patient ventilation without N2O, whether the smallest concentrations of NaHCO3 (1.4% versus 8.4%) reduced diffusion (in vitro evaluation) and other secondary clinical benefits. After pH determination of different solutions (2 mL of 2% L-HCl and 2 to 6 mL of 8.4%, or 1.4% NaHCO3), an in vitro lidocaine diffusion through the ETT cuffs was evaluated (2 mL of 2% L-HCl and 3 mL of 8.4% or 1.4% NaHCO3). Then, adult patients scheduled for total thyroidectomy surgery were consecutively enrolled (n = 20 for each group). The ETT cuff was filled with air (group air) or with alkalinized lidocaine (2 mL of 2% L-HCl) using 8.4% (group large dose) or 1.4% (group small dose) of NaHCO3. After tracheal extubation, sore throat was evaluated by visual analog scale as the main end-point of the study. Hoarseness, bucking, dysphonia, dysphagia, cough, restlessness, and postoperative nausea and vomiting were also evaluated. There was a slight tendency toward a slower release when a small concentration of NaHCO3 was used (i.e., 1.4%). Compared with group air, the alkalinized-lidocaine groups had a significant reduction in sore throat during the 24-h postoperative period (P < 0.0001). The difference was not significant between the two alkalinized lidocaine groups. This increase in ETT tolerance was confirmed by the analysis of secondary end-points. No laryngospasm, rupture of ETT cuff, or depression of the swallowing reflex were recorded. A decrease in sore throat during the postoperative period was recorded when the cuff was inflated with a small dose of alkalinized lidocaine (i.e., 40 mg of L HCl and 1.4% of NaHCO3) rather than with air when ventilation was controlled without N2O. PMID- 16244029 TI - The predictive value of the height ratio and thyromental distance: four predictive tests for difficult laryngoscopy. AB - Preoperative evaluation of anatomical landmarks and clinical factors help identify potentially difficult laryngoscopies; however, predictive reliability is unclear. Because the ratio of height to thyromental distance (RHTMD) has a demonstrably better predictive value than the thyromental distance (TMD), we evaluated the predictive value and odds ratios of RHTMD versus mouth opening, TMD, neck movement, and oropharyngeal view (modified Mallampati). We collected data on 550 consecutive patients scheduled for elective-surgery general anesthesia requiring endotracheal intubation and then assessed all five factors before surgery. An experienced anesthesiologist, not apprised of the recorded preoperative airway assessment, performed the laryngoscopy and grading (as per Cormack and Lehane's classification). Difficult laryngoscopy (Grade 3 or 4) occurred in 69 patients (12.5%). RHTMD had a higher sensitivity, positive predictive value, and fewer false negatives than the other variables tested. In the multivariate analysis, three criteria were found independent for difficult laryngoscopy (neck movement < or =80 degrees; Mallampati Class 3 or 4, and RHTMD > or =23.5). The odds ratio (95% confidence interval) of the RHTMD, Mallampati class, and neck movement were 6.72 (3.29-13.72), 2.96 (1.63-5.35), and 2.73 (1.14 6.51), respectively. The odds ratio for RHTMD was the largest and thus may prove a useful screening test for difficult laryngoscopy. PMID- 16244031 TI - Hypoxemia during one-lung ventilation: jet ventilation of the middle and lower lobes during right upper lobe sleeve resection. AB - A 64-yr-old man underwent right thoracotomy and upper lobectomy for lung carcinoma. Hypoxemia on one-lung ventilation was being managed with continuous positive airway pressure to the nondependent lung when a sleeve resection had to be performed. As this positive airway pressure would no longer be maintained with the bronchus open, an alternate method of oxygenation was necessary. This report describes the successful use of jet ventilation via an airway exchange catheter placed in the bronchus intermedius through the tracheal lumen of a left-sided double-lumen endobronchial tube. Oxygenation was maintained and surgical access was good during the 15-min resection. PMID- 16244032 TI - Report of the 13th annual meeting of the International Society for Anaesthetic Pharmacology. PMID- 16244030 TI - Supplemental intravenous crystalloid administration does not reduce the risk of surgical wound infection. AB - Wound perfusion and oxygenation are important determinants of the development of postoperative wound infections. Supplemental fluid administration significantly increases tissue oxygenation in surrogate wounds in the subcutaneous tissue of the upper arm in perioperative surgical patients. We tested the hypothesis that supplemental fluid administration during and after elective colon resections decreases the incidence of postoperative wound infections. Patients undergoing open colon resection were randomly assigned to small-volume (n = 124, 8 mL.kg( 1).h(-1)) or large-volume (n = 129, 16-18 mL.kg(-1).h(-1)) fluid management. Our major outcomes were two distinct criteria for diagnosis of surgical wound infections: 1) purulent exudate combined with a culture positive for pathogenic bacteria, and 2) Center for Disease Control criteria for diagnosis of surgical wound infections. All wound infections diagnosed using either criterion by a blinded observer in the 15 days after surgery were considered in the analysis. Wound healing was evaluated with the ASEPSIS scoring system. Of the patients given small fluid administration, 14 had surgical wound infections; 11 given large fluid therapy had infections, P = 0.46. ASEPSIS wound-healing scores were similar in both groups: 7 +/- 16 (small volume) versus 8 +/- 14 (large volume), P = 0.70. Our results suggest that supplemental hydration in the range tested does not impact wound infection rate. PMID- 16244035 TI - Anesthetic depth and long-term mortality. PMID- 16244036 TI - Short-lasting effect of perineural resiniferatoxin on mechanical hyperalgesia. PMID- 16244037 TI - Capnogram shape in obstructive lung disease. PMID- 16244038 TI - Hyperkalemic cardiac arrest after cardiopulmonary bypass in a child with Duchenne muscular dystrophy. PMID- 16244039 TI - Countries, rooms, and plants, but not mammalian cells. PMID- 16244040 TI - Sterilization pouches for perioperative thermal insulation. PMID- 16244041 TI - Supplementation of intrathecal bupivacaine with clonidine in ex-premature neonates. PMID- 16244042 TI - Costs are not the only thing we should be concerned with in anesthesia. PMID- 16244043 TI - Airway fires: gas-bugs providing the fuel? PMID- 16244044 TI - Aerosolized lidocaine for relief of extubation laryngospasm. PMID- 16244045 TI - The case for the role of advanced simulators in trauma management training was not made. PMID- 16244046 TI - Anesthesiologists must inform their surgical colleagues when there is a risk of an operating room fire. PMID- 16244047 TI - Thoracotomy in a patient with a history of local anesthetic allergy. PMID- 16244048 TI - Subdural hematoma following accidental dural puncture. PMID- 16244049 TI - ECG artifact and BIS in severe brain injury. PMID- 16244050 TI - The effect of family size on incentive effects of welfare transfers in two-parent families: an evaluation using experimental data. AB - Family size is an important determinant of family well-being, and it is a good predictor of poverty. This study examines effects of waiving the 100-hour rule, by family size, and distinguishes between the "work-incentive effects" and the "eligibility effects" of the waiver. The 100-hour rule limits eligibility to aid to two-parent families in which the principal earner is unemployed or underemployed (works fewer than 100 hours per month). The study uses data from the Link-Up randomized experiment, conducted in California's Central Valley, from 1992 to 1994. The findings show that the eligibility effect of the waiver does not differ by family size, but the work-incentive effect does. PMID- 16244051 TI - Propensity scores: an introduction and experimental test. AB - Propensity score analysis is a relatively recent statistical innovation that is useful in the analysis of data from quasi-experiments. The goal of propensity score analysis is to balance two non-equivalent groups on observed covariates to get more accurate estimates of the effects of a treatment on which the two groups differ. This article presents a general introduction to propensity score analysis, provides an example using data from a quasi-experiment compared to a benchmark randomized experiment, offers practical advice about how to do such analyses, and discusses some limitations of the approach. It also presents the first detailed instructions to appear in the literature on how to use classification tree analysis and bagging for classification trees in the construction of propensity scores. The latter two examples serve as an introduction for researchers interested in computing propensity scores using more complex classification algorithms known as ensemble methods. PMID- 16244052 TI - Eliciting survey cooperation: incentives, self-interest, and norms of cooperation. AB - Given the weaker ties to community as noted by scholars such as Robert Putnam, survey researchers should not be surprised by a decline in survey participation over the past 10 years. This research analyzes the use of incentives coupled with introductory themes emphasizing cooperation and helpfulness--cooperative norms in American society--to understand their effects on survey response. This article analyzes two separate experiments (one phone and one mail) that provide evidence that norms of cooperation matter in the decision to participate in a survey, suggesting that this is particularly true at the refusal conversion stage. These results indicate that survey researchers may use such themes to their advantage, especially when conducting a nonresponse follow-up in a mail survey. PMID- 16244053 TI - Training x Trainee Interactions in a family planning intervention. AB - This study investigated the effects of introducing a family planning counseling model at clinics of Peru's Ministry of Health. Providers trained in the model presented greater quality of care and longer counseling sessions than did controls. The main effects, however, were misleading. Nearly all of the quality improvements were contributed by 37% of the trained providers; they had already been better performers at the pretest and complied with the new counseling model's requirement of job aid use at the posttest. The recognition that a majority of trained providers did not profit from training posed a challenge that led to improvements in the model. Training x Trainee Interactions should be explored in evaluations. PMID- 16244054 TI - The learning evaluation: a theoretical and empirical exploration. AB - In this article, the authors theoretically and empirically explore the concept of learning evaluation. They shed light on the positioning of the learning evaluation amid scholarly work on evaluations. Moreover, they describe the learning evaluation in practice in the Netherlands by going into a specific project called the Stimulation Program on Citizen and Environment. The theoretical and empirical quest gives insights into the problems with and possibilities of the learning evaluation. They think that their experiences can help the further development of theory about learning evaluation as well as aid in the practice of such evaluations. PMID- 16244056 TI - Long-term dynamics of Sin Nombre viral RNA and antibody in deer mice in Montana. AB - Infections with hantaviruses in the natural host rodent may result in persistent, asymptomatic infections involving shedding of virus into the environment. Laboratory studies have partially characterized the acute and persistent infection by Sin Nombre virus (SNV) in its natural host, the deer mouse (Peromyscus maniculatus). However, these studies have posed questions that may best be addressed using longitudinal studies involving sequential sampling of individual wild-caught, naturally infected mice. Using enzyme immunoassay and polymerase chain reaction (PCR) analysis of monthly blood samples, we followed the infection status of deer mice in a mark-recapture study in Montana for 2 yr. Only six of 907 samples without IgG antibody to SNV contained detectable SNV RNA, suggesting that there is a very brief period of viremia before the host develops detectable antibody. The simultaneous presence of both antibody and viral RNA in blood was detected in consecutive monthly samples for as long as 3 mo. However, chronic infection was typified by alternating characteristics of PCR positivity and PCR negativity. Two possible interpretations of these results are that 1) viral RNA may be consistently present in the blood of chronically infected deer mouse, but that viral RNA is near the limits of PCR detectability or 2) SNV RNA sporadically appears in blood as a consequence of unknown physiological events. The occurrence of seasonal patterns in the proportion of samples that contains antibody and that also contained SNV RNA demonstrated a temporal association between recent infection (antibody acquisition) and presence of viral RNA in blood. PMID- 16244057 TI - PCR detection of bovine herpesviruses from nonbovine ruminants in Hungary. AB - Polymerase chain reaction (PCR) was used to test six different nonbovine ruminant species for five bovine herpesviruses including infectious bovine rhinotracheitis virus (BoHV-1), bovine herpes mammillitis virus (BoHV-2), Movar-type herpesvirus (BoHV-4), bovine herpesvirus type 5 (BoHV-5), and alcelaphine herpesvirus 1 (AlHV 1). Species tested included 56 roe deer (Capreolus capreolus), 66 red deer (Cervus elaphus), 20 fallow deer (Dama dama), 16 mouflon (Ovis musimon), 34 domestic sheep, and 44 domestic goats, which were sampled in Hungary in 2003. Tracheal and popliteal lymph nodes collected from these animals were tested for the presence of the five bovine herpesviruses using three nested (two of which were duplex) PCR assays. Three bovine herpesviruses (BoHV-1, -2, and -4) were detected, whereas no evidence of AlHV-1 or BoHV-5 was observed. Prevalence of BoHV-1 ranged from 12% to 47%, and PCR-positive results were observed in all species tested. BoHV-2 was detected from roe deer, red deer, fallow deer, mouflon, and domestic sheep, and the prevalence in these species ranged from 3% to 50%. BoHV-4 was detected in all species, with prevalence ranging from 12% to 69%. Sequenced PCR products were 99-100% identical to bovine herpesviral sequences deposited in the GenBank. PMID- 16244058 TI - Distribution of chelonid fibropapillomatosis-associated herpesvirus variants in Florida: molecular genetic evidence for infection of turtles following recruitment to neritic developmental habitats. AB - Marine turtle fibropapillomatosis is associated with chelonid fibropapilloma associated herpesvirus (C-FP-HV) and commonly affects juvenile green turtles (Chelonia mydas) in neritic (nearshore) habitats. Green turtles have a complex life history, characterized by shifts in trophic level as well as habitat during ontogeny. Thus, several hypotheses can be proposed for when turtles become infected with C-FP-HV. They may acquire the virus at an early stage in the life cycle, including prenatal, hatchling, or the posthatchling pelagic stages. Alternatively, they may become infected later in life after they emigrate from the open ocean to neritic habitats. Each hypothesis generates predictions about the spatial distribution of genetic variants of C-FP-HV among nearshore sites within a region. Sequencing of polymerase chain reaction-amplified viral DNA from fibropapillomas of individual turtles was used to genotype the viral variants present in marine turtles from different coastal areas in Florida. We found four distinct virus variants (A, B, C, and D), two of which (A and C) were present in multiple turtle species. Green turtles in Florida were infected with variants A, B, and C. Variant A was found in green turtles from all three areas. Outside the Indian River Lagoon, variant A was most commonly detected and was found in >94% of diseased green turtles and 70% of loggerhead sea turtles (Caretta caretta) in the Florida Bay/Florida Keys. However, in the Indian River Lagoon, variant B was found in >94% of affected green turtles. Variant B was not detected outside of the Indian River system. Chi-square analysis strongly supported (P<0.001) an association between viral variant distribution in green turtles and location. On the basis of the assumption that juvenile green turtles found in Florida's west central coast, Florida Keys, and Indian River Lagoon areas represented recruits from a mixed pelagic population, we expected that the distribution of viral variants in these turtles would be relatively homogeneous among locations; this would correspond to infection in the earlier phases of their life cycle. The heterogeneous distribution of viral variants in green turtle tumors from different Florida coastal locations strongly supports the hypothesis that, during epizootics, turtles are infected with specific C-FP-HV variants after they arrive as juveniles in neritic habitats. The conclusion that C-FP-HV is acquired after turtles recruit to nearshore habitats should help focus further research efforts on understanding the mechanisms of transmission and raises the possibility that the effect of fibropapillomatosis on turtle populations might be reduced by management strategies designed to break the cycle of transmission in these locations. PMID- 16244059 TI - Avian cholera exposure and carriers in greater white-fronted geese breeding in Alaska, USA. AB - We conducted a 3-yr study (2001-03) on greater white-fronted geese (Anser albifrons frontalis) breeding in Alaska, USA, to determine the exposure of this population to Pasteurella multocida and the potential role of these birds as disease carriers. We tested sera from nearly 600 adult geese for antibodies to P. multocida serotype 1. We found a low prevalence (<5%) of positive antibodies in adult geese, and based on the short duration of detectable antibodies, these findings indicate recent infection with P. multocida. Prevalence was similar to serologic results from both breeding and wintering lesser snow geese. We also collected oral (n=1,035), nasal (n=102), and cloacal (n=90) swab samples to determine the presence of avian cholera carriers in this population. We were unable to isolate P. multocida serotype 1 from any of the birds sampled. Based on comparison with other waterfowl species, we concluded that these geese may be exposed to avian cholera during the winter or spring migration but are unlikely to play a significant role as carriers of the bacterium causing avian cholera. PMID- 16244060 TI - Relative vulnerability of chronic wasting disease infected mule deer to vehicle collisions. AB - We estimated chronic wasting disease (CWD) prevalence among vehicle-killed mule deer (Odocoileus hemionus) in select data analysis units (DAUs) in northern Colorado, USA, and compared these with estimated CWD prevalence among mule deer of the same sex sampled in the vicinity of collision sites to assess relative vulnerability of CWD-infected individuals to vehicle collisions. Twenty-five of 171 vehicle-killed mule deer tested positive for CWD (overall prevalence=0.146, 95% confidence interval [CI]=0.097-0.208); 173 of 2,317 deer sampled in the vicinity of these vehicle-killed deer tested positive (overall prevalence=0.075, 95% CI=0.064-0.085). In nine of ten DAU x sex comparisons, relative risk of CWD infection tended to be higher among vehicle-killed deer (range of estimated relative risks=1.6-15.9). Spongiform encephalopathy was detected in 12 of 20 (60%; 95% CI=39-81%) CWD-positive deer killed by vehicles and in 79 of 180 (44%; 95% CI=37-52%) CWD-positive deer detected via random sampling (relative risk=1.37; 95% CI=0.92-2.03), suggesting that infected deer killed by vehicles tended to be in later stages of disease than those killed by hunters. Our data offer evidence that CWD-infected mule deer may be relatively vulnerable to vehicle collisions. It follows that sampling of vehicle-killed mule deer may be exploited to increase efficiency of surveillance programs designed to detect new foci of CWD infection; moreover, evidence of increased susceptibility to vehicle collisions may aid in understanding vulnerability of CWD-infected individuals to other forms of death, particularly predation. PMID- 16244061 TI - Infectious disease and the decline of Steller sea lions (Eumetopias jubatus) in Alaska, USA: insights from serologic data. AB - Serologic data were examined to determine whether infectious disease may have played a role in the decline of Steller sea lions (Eumetopias jubatus) in the Gulf of Alaska and Aleutian Islands, USA. Available published data, unpublished data, and recent collections (1997-2000) were compared and reviewed. Data were stratified by geography to compare the declining western Alaskan population in the Aleutian Islands through eastern Prince William Sound to the increasing population in southeastern Alaska. Prevalences of antibodies from the 1970s to the early 1990s were noted for Leptospira interrogans, Chlamydophila psittaci, Brucella spp., phocid herpesvirus-1, and calciviruses. Serum samples collected from 1997-2000 were tested for antibodies to these agents as well as to marine mammal morbilliviruses, canine parvovirus, and canine adenovirus-1 and -2. Conclusions could not be drawn about changes in antibody prevalence to these agents during the decline of Steller sea lions, however, because data were incomplete or not comparable as a result of inconsistencies in testing techniques. Despite these shortcomings, results provided no convincing evidence of significant exposure of Steller sea lions to morbilliviruses, Brucella spp., canine parvovirus, or L. interrogans. Steller sea lions have been exposed to phocid herpesviruses, caliciviruses, canine adenovirus, and C. psittaci or to cross-reactive organisms in regions of both increasing and decreasing sea lion abundance. Based on similar antibody prevalence estimates from the increasing and decreasing populations, these agents are unlikely to have been the primary cause of the population decline. They may have contributed to the decline or impeded population recovery, however, because of undetected mortality and morbidity or reductions of fecundity and body condition in animals under other stresses. Systematic monitoring for disease agents and their effects is needed to determine whether infectious disease currently plays a role in the decline and lack of recovery of Steller sea lions. PMID- 16244062 TI - The rise and fall of psoroptic scabies in bighorn sheep in the San Andres Mountains, New Mexico. AB - Between 1978 and 1997, a combination of psoroptic scabies (Psoroptes spp.), mountain lion (Puma concolor) predation, and periodic drought reduced a population of native desert bighorn sheep (Ovis canadensis) in the San Andres Mountains (SAM), New Mexico, from >200 individuals to a single ewe. In 1999, this ewe was captured, ensured to be Psoroptes-free, and released back into the SAM. Eleven radio-collared rams were translocated from the Red Rock Wildlife Area (RRWA) in New Mexico into the SAM range and monitored through 2002 to determine whether Psoroptes spp. mites were still in the environment. None of these sentinel rams acquired scabies during this period, and no additional native sheep were found to be present in the range. In 2002, 51 desert bighorn sheep were translocated into the SAM from the Kofa National Wildlife Refuge in Arizona (n = 20) and the RRWA in New Mexico (n = 31). Twenty-one bighorn sheep have died in the SAM since that time, but Psoroptes spp. mites have not been detected on any of these animals, nor have they been found on mule deer (Odocoileus hemionus) sampled since 2000. We conclude that psoroptic scabies is no longer present in the San Andres bighorn sheep population and that psoroptic scabies poses a minimal to nonexistent threat to the persistence of this population at this time. PMID- 16244063 TI - Interspecific differences in hematozoan infection in Sonoran desert Aimophila sparrows. AB - Numerous studies have identified factors that control avian hematozoan infections, but the mechanisms that account for host differences in parasitemia remain largely speculative. To address this issue, we compared the prevalence of these parasites in stained blood smears from four conspecific Sonoran desert Aimophila sparrow species sampled during their breeding season: rufous-winged (Aimophila carpalis; RWSP), rufous-crowned (Aimophila ruficeps; RCSP), Cassin's (Aimophila cassinii, CASP), and Botteri's (Aimophila botterii; BOSP) sparrows. Blood smears contained Haemoproteus fringillae (RWSP), Trypanosoma everetti (RWSP, RCSP, BOSP), Trypanosoma avium (CASP), and microfilariae (all species). Most (92.5%) RWSP (n=40) were infected with Haemoproteus, but this parasite was not detected in RCSP (n=20) or BOSP (n=20) and was found only in one (2.5%) CASP (n=40). Trypanosoma spp. and microfilariae were detected in all species, but prevalence differed between these four sparrow species. Species differences in parasite prevalence were not due to difference in sex, age, adult body mass, incubation period, breeding habitat, or plumage colorfulness. However, differences in Haemoproteus sp. prevalence correlated with preferred nesting height, as RWSP generally nest above ground, whereas the other species nest on or close to the ground. Elevated H. fringillae prevalence in breeding-condition RWSP presumably does not result from a seasonal relapse associated with breeding or require new infection because 1) this prevalence did not differ in males sampled during and outside (n=21) the breeding season, and 2) all male RWSP (n=25) that we held in captivity and shielded from new infections and influence of natural photoperiod for 1 yr had viable blood H. fringillae gametocytes. H. fringillae prevalence in fall-sampled hatch-year male RWSP (n=11) was 63.6%, demonstrating that this parasite can be transmitted on the breeding grounds and during the first months of life. T. everetti prevalence in RWSP was lower in winter than in summer and also in long-term captive than in free-ranging adults. Presence of this parasite in the blood of breeding males may depend on recrudescence of existing infections or new infections. PMID- 16244064 TI - Extrinsic factors significantly affect patterns of disease in free-ranging and captive cheetah (Acinonyx jubatus) populations. AB - The cheetah (Acinonyx jubatus) has been considered a paradigm for disease vulnerability due to loss of genetic diversity. This species monomorphism has been suspected to be the basis for their general poor health and dwindling populations in captivity. North American and South African captive populations have high prevalences of hepatic veno-occlusive disease, glomerulosclerosis, gastritis, and systemic amyloidosis, diseases that are rare in other species. Unusually severe inflammatory reactions to common infectious agents have also been documented in captive cheetahs. The current study compared disease prevalences in free-ranging Namibian cheetahs with those in two captive populations of similar ages. The occurrence of diseases in the free-ranging population was determined from 49 necropsies and 27 gastric biopsies obtained between 1986 and 2003 and compared with prevalences in 147 North American and 80 South African captive cheetahs. Except for two cheetahs, the free-ranging population was in robust health with only mild lesions present, in contrast with significantly higher prevalences in the captive populations. Despite widespread heavy Helicobacter colonization in wild cheetahs, only 3% of the free-ranging population had moderate to severe gastritis, in contrast with 64% of captive cheetahs. No severe inflammatory reactions to viral infections were detected in the free-ranging animals. Because free-ranging Namibian cheetahs are as genetically impoverished as captive cheetahs, these findings caution against attributing loss of fitness solely to genetic factors and attest to the fundamental importance of extrinsic factors in wildlife health. PMID- 16244065 TI - Evaluation and significance of tetracycline stability in rabies vaccine baits. AB - Tetracycline is widely used as a biomarker for bait consumption by wildlife; tetracycline is incorporated into bones and teeth and can be detected by fluorescence microscopy several weeks postconsumption. During 2003, the United States Department of Agriculture distributed more than 10 million tetracycline containing rabies-vaccine baits to control the spread of wildlife vectored rabies to humans, pets, and livestock. To estimate the percentage of target species consuming the baits, raccoons and skunks were collected in baited areas and teeth were analyzed for the presence of the biomarker. Several incidents of low biomarker detection rates prompted an investigation of the stability of the biomarker in the baits. Baits were collected at several points along the manufacturing and distribution chain. Baits were analyzed for free and polymer bound tetracycline and the less active isomer epitetracycline. Results indicated that a portion of the tetracycline was converted to epitetracycline. Additionally, significant quantities of both compounds were trapped in the polymer, which is homogeneously distributed throughout the bait. The results of this study suggest that approximately 40% of the target quantity of tetracycline was unavailable for absorption. This situation could contribute to low biomarker detection rates and suggests that formulation modification should be considered. PMID- 16244067 TI - Liver concentrations of copper, cobalt, and selenium in wild Norwegian red deer (Cervus elaphus). AB - Liver samples from 245 wild red deer (Cervus elaphus) collected during the licensed hunting season in 2001 from five different locations in western Norway were analyzed for copper (Cu), cobalt (Co), and selenium (Se). The associations between these trace elements and geographical location, age group, and sex were studied. The median (and range of) liver concentrations (microg/g wet weight) for all the examined deer were: Cu 20 (1.7-103), Co 0.08 (<0.01-0.18), and Se 0.09 (0.04-1.0). The results indicate a generally low status of Cu and Se. In total, 15 (6%) red deer had deficient Cu levels (< 4 (microg/g). For all three elements, the liver concentrations showed a significant geographic variation. The geographic difference was most distinct for Cu. The lowest median Cu concentration was found in deer from the island Hitra, where 13% of the animals had deficient Cu levels. Significant differences between age groups were found for all elements, and generally, the adults (> or =2.5 yr) had the highest levels. No significant sex differences within the various age groups were found, with three exceptions: female calves and adults had significantly higher Co levels than male deer, and adult males had significantly higher Se levels than adult females. The Cu and Se status of wild red deer in parts of Norway is low; however, the significance of this needs to be explored further. PMID- 16244066 TI - Determination and evaluation of an optimal dosage of carfentanil and xylazine for the immobilization of white-tailed deer (Odocoileus virginianus). AB - Using an iteration method, optimal hand-injected immobilization dosages of carfentanil/xylazine (CAR/XYL) were determined for 13 adult white-tailed deer (Odocoileus virginianus). Deer were temporarily restrained in a squeeze chute and were repeatedly immobilized one to four times at 2-5-wk intervals from December 2002 to March 2003. A fixed ratio of 1 mg CAR:10 mg XYL intramuscularly was used, increasing or decreasing the dosage until the optimal dosage (defined by an induction time < 3 min and PaCO(2)< 60 mmHg) was reached for each animal. Inductions were video-recorded and reviewed by observers blinded to drugs and dosages, who rated qualitative aspects of each induction. There were significant (P < 0.05) dosage-dependent decreases in induction time, time to first effect, PaO(2), SaO(2), and arterial pH, and significant dosage-dependent increases in PaCO(2) and quality ratings. The median optimal dosage (mOD) was 0.03 (range, 0.015-0.06) mg/kg CAR+0.3 (range, 0.15-0.6) mg/kg XYL. Induction times using the mOD were rapid (median 3.0 min [range, 1.8-10.0]), but quality ratings were considered undesirable for nine of 13 deer. Increased rectal body temperatures of 40.6+/-0.5 C (mean +/- SD) were noted in all deer and hyperthermia (T > 41 C) was noted in three. There was a positive correlation between body temperature and induction time (r=0.44). Heart rates significantly decreased from 5 to 15 min postinduction and remained decreased at the 20-min reading; there was occasional bradycardia. There was a significant increase in pH from 10 to 20 min postinduction, but metabolic acidemia (pH<7.3) persisted throughout the immobilization periods for all deer. Possible hypoxemia (SaO(2) and SpO(2)<90 mmHg but PaO(2)>60 mmHg) was present after induction, while hypercapnea (PaCO(2) > 60 mmHg) did not occur. Reversal times with naltrexone and yohimbine were rapid (mean 3.7+/-1.5 min) and uneventful, with no evidence of renarcotization. Although the median optimal dosage produced rapid inductions, no respiratory depression, complete reversal after antagonist administration, and no renarcotization, negative attributes included elevated body temperatures, acidemia, and undesirable induction qualities. PMID- 16244068 TI - Further observations on the blood parasites of birds in Uganda. AB - Birds from three National Parks (Bwindi Impenetrable, Kibale, and Queen Elizabeth) in western Uganda were surveyed during the dry season in July 2003 and investigated for hematozoa by microscopic examination of stained blood films. Of 307 birds examined, representing 68 species of 15 families and four orders, 61.9% were found to be infected with blood parasites. Species of Haemoproteus (15.3% prevalence), Plasmodium (20.5%), Leucocytozoon (40.1%), Trypanosoma (11.4%), Hepatozoon (2.6%), Atoxoplasma (0.3%), and microfilariae (3.9%) were recorded. Except for Haemoproteus spp. infections, the overall prevalence of hematozoa belonging to all genera was significantly higher in this study than was previously reported in Uganda. Thirty-six species of birds were examined for blood parasites for the first time and 112 new host-parasite associations were identified. Eighty-one were at the generic and 31 at the specific level of the hematozoa. Hepatozoon and Atoxoplasma spp. were detected for the first time in Uganda. PMID- 16244069 TI - Experimental infection of adult and juvenile coyotes with domestic dog and wild coyote isolates of Hepatozoon americanum (Apicomplexa: Adeleorina). AB - Each of five adult and four juvenile coyotes (Canis latrans) was exposed to an oral dose of 50 Hepatozoon americanum oocysts recovered from Amblyomma maculatum ticks that previously fed on either naturally infected domestic dogs (Canis familiaris) or naturally infected wild coyotes. All coyotes exposed to H. americanum became infected, regardless of isolate source, and all exhibited mild to moderate clinical disease that simulated American canine hepatozoonosis in naturally infected dogs. At 100 days postexposure, parasitemia was greater in juvenile than adult coyotes (0.9% and 0.3%, respectively); radiographic imaging of femurs revealed moderate exostosis in all juveniles and mild to moderate new bone growth in four of five (80%) adult coyotes. Gross postmortem analysis of bone lesions demonstrated variation between age groups of coyotes but not between isolates of H. americanum. Microscopic evaluation of skeletal muscle revealed that parasite-induced lesions were significantly more numerous (t = 5.0, df = 7, P = 0.001) in juvenile than adult coyotes. Results of this study indicate that juvenile and adult coyotes are equally susceptible to experimental infection with H. americanum isolated from domestic dog and wild coyote sources. The age of coyotes at the time of exposure, and possibly the number of H. americanum oocysts ingested, might influence morbidity and mortality, but it appears that both adult and juvenile coyotes could be reservoirs of H. americanum. PMID- 16244070 TI - Metastrongyloid nematode (Otostrongylus circumlitus) infection in a stranded California sea lion (Zalophus californianus)--a new host-parasite association. AB - A stranded yearling male California sea lion was admitted to a rehabilitation center June 2003. On presentation, the sea lion was emaciated and had diarrhea and neutrophilia. Two weeks later, the animal became anorexic, blood and mucus were observed around the oral cavity, and corneal opacity was noted in the right eye. Hematology results at that time included leukocytosis consisting of neutrophilia with a left shift, anemia, and thrombocytopenia. Despite supportive care, the sea lion died. On post mortem examination, there were multiple areas of hemorrhage scattered throughout all lung lobes, and pulmonary blood vessels were occluded by fibrin thrombi. Nematodes identified as immature forms of Otostrongylus circumlitus were found in the right ventricle and pulmonary arteries. Histologic findings in the lungs included severe suppurative and necrotizing arteritis with vascular thrombosis, interstitial pneumonia, and large areas of pulmonary hemorrhage. This report of O. circumlitus infection in a California sea lion (Zalophus californianus) might indicate a potentially new host-parasite association. PMID- 16244071 TI - Absence of antibodies to foot-and-mouth disease virus in free-ranging roe deer from selected areas of Germany (2001-2002). AB - Blood samples (n = 223) of free-ranging roe deer (Capreolus capreolus) were collected from selected hunting grounds in Germany between October 2001 and October 2002. Samples originated from Lower Saxony (n = 43) and North-Rhine Westphalia (n = 108) within a 20-km area ("cordon") cordoned off along the border of The Netherlands. This is adjacent to the area of a foot-and-mouth disease outbreak that occurred between 21 March and 22 April 2001 in The Netherlands. Negative control samples were taken from northern Germany (Schleswig-Holstein, n = 72). Two different enzyme-linked immunosorbent assays (ELISAs) were used for the detection of antibodies against foot-and-mouth disease virus (FMDV) serotype O strain Manisa. To confirm ELISA-positive results, a virus neutralization test was performed. All samples tested negative for antibodies against FMDV. These results suggest that FMDV was not transmitted to free-ranging roe deer living in parts of Germany adjacent to the area affected by the 2001 foot-and-mouth disease outbreak in The Netherlands. PMID- 16244072 TI - Detection of a newly described pestivirus of Pyrenean chamois (Rupicapra pyrenaica pyrenaica) in France. AB - A pestivirus was detected and characterized in chamois (Rupicapra pyrenaica pyrenaica) originating from the French part of the Pyrenees. Phylogenetic analysis of the pestivirus was done on the basis of a fragment from the 5' noncoding region including 22 published nucleotide sequences of different pestivirus strains. Our strain was grouped within the clade of border disease viruses (BDV). However, it had an intermediate position between clade BDV and classical swine fever viruses representing a basal position to BDV strains of domestic sheep. Our strain was grouped as a sister unit to a novel pestivirus (Chamois-1) recently described from chamois in Spain. Therefore, we postulate that this virus occurs in the entire population of Pyrenean chamois. On the basis of the phylogenetic grouping of this isolate, a postulated cross-species transmission of pestivirus from domestic sheep to chamois via shared pastures seems to be unlikely. PMID- 16244073 TI - Antibodies to the Ross River virus in captive marsupials in urban areas of eastern New South Wales, Australia. AB - Serum samples collected from 224 tammar wallabies (Macropus eugenii) in two captive populations in urban areas in eastern New South Wales Australia, between December 1999 and May 2004, were tested for antibodies to Ross River virus (RRV). In one population in northwest Sydney, 21 animals (11%) tested positive, and in another population in Newcastle, New South Wales, thirteen (33%) of the animals were positive. Antibodies were detected in four of 11 wallaroos (Macropus robustus) (36%) but not in parma wallabies (Macropus parma) (n=5), koalas (Phascolarctos cinereus) (n=12) and southern hairy-nosed wombats (Lasiorhinus latifrons) (n=2) from the Sydney area. These data support the possible role of marsupials as urban amplifying hosts for RRV. PMID- 16244074 TI - A serological and fecal parasitologic survey of the critically endangered pygmy raccoon (Procyon pygmaeus). AB - The pygmy raccoon (Procyon pygmaeus) of Cozumel Island, Mexico, is among the most endangered carnivores in the world, and causes of its decline are unclear. During 2002 and 2003, we sampled approximately 10% of the remaining population to survey exposure to viral and parasitic pathogens that may have contributed to population decline. We found evidence of exposure to infectious canine hepatitis, canine distemper, feline panleukopenia virus, and Toxoplasma gondii. The latter is suggestive of spillover from domestic cats, which have only recently been introduced to the island. Additional parasites identified include Eimeria nutalli, Placoconus lotoris, Capillaria procyonis, Physaloptera sp., a mite in the family Listrophoridae, and a trematode in the family Heterophyidae. Several of these are typical of the parasite community of the common raccoon (Procyon lotor). PMID- 16244075 TI - Highly pathogenic avian influenza in magpies (Pica pica sericea) in South Korea. AB - Highly pathogenic avian influenza (HPAI) is an extremely infectious, systemic viral disease of birds that produces high mortality and morbidity. HPAI was diagnosed in the three dead magpies (Pica pica sericea) submitted to the National Veterinary Research and Quarantine Service. At necropsy, the prominent lesions were multifocal or coalescing necrosis of the pancreas with enlargement of the livers and spleens. Microscopically, there were severely necrotizing pancreatitis and lymphocytic meningoencephalitis. Influenza viral antigen was also detected in areas closely associated with histologic lesions. Avian influenza virus was isolated from cecal tonsils and feces of the magpies. The isolated virus was identified as a highly pathogenic H5N1, with hemagglutinin proteolytic cleavage site deduced amino acid sequence of QREKRKKR/GLFGAIAG. To determine the pathogenicity of the isolate, eight 6-wk-old specific-pathogen-free chickens were inoculated intravenously with the virus, and all birds died within 24 hr after inoculation. This is the first report of HPAI in magpies. PMID- 16244076 TI - Avian influenza viruses and paramyxoviruses in wintering and resident ducks in Texas. AB - Cloacal swabs were collected from teal (Anas crecca, Anas cyanoptera, Anas discors), mottled duck (Anas fulvigula) and northern pintail (Anas acuta) in Brazoria County, Texas, USA, during February 2001, mottled ducks during August 2001, and blue-winged teal (A. discors) during February 2002. Prevalence of avian influenza virus (AIV) infections during each sampling period were 11, 0, and 15%, respectively. The hemagglutinin (H) subtypes H2 and H7 were detected in both years, while the H8 subtype was detected in 2001 and the H1 subtype was detected in 2002. Avian paramyxovirus type 1 (APMV-1) was isolated from 13% of mottled ducks sampled in August 2001 and 30.7% of teal in February 2002. The season of isolation of both viruses and the majority of the AIV subtypes detected in this study are not typical based on previous reports of these viruses from North American ducks. PMID- 16244077 TI - Isolation of Mycobacterium avium subsp paratuberculosis (Map) from feral cats on a dairy farm with Map-infected cattle. AB - Paratuberculosis is an economically important disease of dairy cattle caused by Mycobacterium avium subsp. paratuberculosis (Map). The role of nonruminant, nondomestic animals in the epidemiology of paratuberculosis in cattle is unclear. To examine nonruminant, nondomestic animals for the presence of Map, 25 feral cats, nine mice (species unknown), eight rabbits (Sylvilagus floridanus), six raccoons (Procyon lotor), and three opossums (Didelphis virginiana) were collected from a mid-western dairy with known Map-infected cattle. Mycobacterium avium subsp. paratuberculosis was isolated from the mesenteric lymph node from seven of 25 (28%) feral cats. Ileum was culture-positive for three of these seven cats, and an isolation of Map was also made from the ileum of one of nine (11%) mice. Tissue samples from other species were negative as determined by Map culture; microscopic lesions consistent with paratuberculosis were not seen in any animal. Restriction fragment polymorphism analysis of isolates from cats and dairy cattle suggest interspecies transmission. The means by which interspecies transmission occurred may be through ingestion of Map-contaminated feces or waste milk or through ingestion of Map-infected prey. Shedding of Map from infected cats was not evaluated. The epidemiologic role of Map-infected feral cats on dairy farms requires further investigation. PMID- 16244079 TI - Hailstorms as a cause of mass mortality of Swainson's hawks in their wintering grounds. AB - We describe a mass mortality event of wintering Swainson's hawks (SH, Buteo swainsoni) in central Argentina during November 2003. One hundred thirteen SH were found dead as a consequence of a single hailstorm. In addition, 14 hawks with severe injuries were recovered alive, but only 10 of these survived 1 wk later. Another 45 dead birds of 11 species were collected in the area. Interviews with local land-owners conducted in other areas of the SH wintering grounds provided further evidence of past hailstorm-related mortality involving SH, suggesting that such events commonly occur in the Argentine Pampas. This potential cause of mass mortality of SH wintering in agricultural areas of Argentina may be significant when added to the increased mortality associated with poisoning events during the last decade. PMID- 16244078 TI - Presumptive gangrenous ergotism in free-living moose and a roe deer. AB - Presumptive gangrenous ergotism in 10 moose (Alces alces) and one roe deer (Capreolus capreolus) is reported. Three of the moose came from a municipality in southeastern Norway where the disease occurred as a cluster in 1996. The other moose represented solitary or sporadic cases diagnosed in four municipalities in northwestern Norway between 1996 and 2004. Affected moose (seven calves, three yearlings) were found between October and June, showing distal limb lesions on one to three limbs. The lesions in the moose found during October and November presented as dry gangrene, whereas moose found between December and June presented with loss of the distal part of the limbs or open lesions close to sloughing. Four of the moose also had bilateral ear lesions affecting the outer third of the pinnae. A retrospective diagnosis of ergotism (June 1981) was made in a 1-yr-old roe deer from northwestern Norway showing loss of the distal part of all four limbs. PMID- 16244080 TI - Epidemic trichodinosis associated with severe epidermal hyperplasia in largemouth bass, Micropterus salmoides, from North Carolina, USA. AB - An epidemic of trichodinosis associated with severe epidermal hyperplasia occurred in adult largemouth bass (Micropterus salmoides) from the Chowan River drainage, North Carolina (USA) in late winter to early spring 2002. Initial reports by anglers of fish with a "jelly-like slime coat" on the skin prompted an electrofishing survey in which about 10% of sampled largemouth bass had a very thick, bluish-white "mucoid layer" on the body and fins. Moderate to heavy infestations of the ciliate Trichodina were detected in wet mounts of skin from five of five fish having the mucoid layer; these fish also had significant gill infestations. An additional two fish with only mild reddening and four asymptomatic fish (no skin lesions) had mild skin infestations but no gill infestations. Two asymptomatic fish had no skin parasites. Four fish with the mucoid layer were necropsied and had extremely severe epidermal hyperplasia on the body and fins. The hyperplasic epidermis had relatively few mucus cells and typically was about 5-10 times thicker than healthy epidermis. The upper four fifths of the epidermis consisted of finely vacuolated, highly flattened, somewhat disorganized epithelial cells. No other significant clinical or histopathologic abnormalities were detected. No systemic infection by pathogenic bacteria was noted. The environmental cause of the epidemic is uncertain but the lesions suggest that some chronic stressor was involved. PMID- 16244081 TI - Congenital hemicerebral anomaly in a stranded Pacific harbor seal (Phoca vitulina richardsi). AB - A stranded 5-month-old female Pacific harbor seal (Phoca vitulina richardsi) was presented displaying tachypnea and diminished lung sounds. No neurological abnormalities were noted. The animal was treated for verminous pneumonia, but died 2 wk later. Gross necropsy examination revealed a severe obstructive verminous pneumonia associated with large numbers of Otostrongylus circumlitus. In addition, the majority of the right cerebral hemisphere was absent, with hypoplasia of the left cerebellar hemisphere, absence of the right pyramid, and malformation of the right occipital bone. Histopathologic findings included multifocal thrombosis and inflammation of pulmonary arteries, verminous pneumonia, and mild vacuolation of the subependymal white matter in the third ventricle representing swelling of myelin sheaths and edema. This is the first report of a hemicerebral anomaly in a marine mammal. PMID- 16244082 TI - Tiletamine-zolazepam-xylazine immobilization of American marten (Martes americana). AB - The effectiveness of tiletamine-zolazepam (Telazol) and xylazine as an immobilizing combination for American martens (Martes americana) was evaluated. Fifteen martens were intramuscularly injected on 19 occasions using a 3:2 mixture of tiletamine-zolazepam (3.2+/-0.6 mg/kg [mean +/- SD]) and xylazine (2.1+/-0.4 mg/kg) at Pictured Rocks National Lakeshore, Michigan (USA) during May to October 2002-2003. Mean induction time was 2.5+/-1.8 min; mean recovery time was 70.8+/ 31.9 min. There was no relation between the amount (mg/kg) of tiletamine zolazepam-xylazine injected and induction (r(2)=0.08, P=0.26). However, there was an inverse relation (r(2)=0.28, P<0.01) between dosage and time to first effect of immobilants. Time to recovery increased (r(2)=0.21, P=0.05) with increased dosage. Mean heart rate, respiratory rate, and body temperature declined through 10 min postinduction (P<0.05). No mortality occurred and no short-term adverse effects were observed in recaptured individuals. In conclusion, a 3:2 mixture of tiletamine-zolazepam/xylazine is a safe and effective immobilizing agent for martens when conducting non-surgical field procedures. Immobilizing martens with 4.2 mg/kg tiletamine-zolazepam and 2.8 mg/kg xylazine should provide < or =30 min of handling time and allow full recovery in about 70 min. PMID- 16244083 TI - Thallium contamination in wild ducks in Japan. AB - Although thallium (Tl) is toxic to both humans and animals, there is little information on contamination in wildlife. In this study, Tl contents in wild ducks in Japan were determined. Contents of Tl in kidney and liver ranged from 0.42 to 119.61 and 0.10 to 33.94 microg/g dry weight, respectively. Significant correlations between Tl contents in kidney and liver were observed for all dabbling ducks except mallard (Anas platyrhynchos); similar correlations were not observed in diving ducks. Variation in Tl content was observed between sampling locations with the highest mean Tl content in the Eurasian wigeon (Anas penelope) collected in Ibaraki Prefecture. PMID- 16244085 TI - Linezolid use in sepsis due to methicillin-susceptible Staphylococcus aureus. PMID- 16244084 TI - Surveillance of extended-spectrum beta-lactamases from clinical samples and faecal carriers in Barcelona, Spain. AB - OBJECTIVES: The aim of the present study was to characterize and compare the extended-spectrum beta-lactamase (ESBL)-producing organisms isolated from clinical samples and faecal carriers in 2001 and 2002. METHODS: A total of 5251 Enterobacteriaceae isolated from clinical samples and 1321 stool samples were evaluated for the presence of ESBLs. The stool samples were spread onto plates of MacConkey agar containing 2 mg/L cefotaxime for selection of ESBL-producing strains. These strains were defined as those showing synergism between amoxicillin/clavulanic acid and third-generation cephalosporins. The beta lactamases involved were characterized by isoelectric focusing, PCR assays and DNA sequencing. RESULTS: The prevalence of ESBL-producing strains among clinical Enterobacteriaceae was 1.7%. Of these, 87.6% produced CTX-M, 25.8% produced SHV and 2.2% were TEM-type-producing strains. All clinical ESBL-producing strains were Escherichia coli, with the exception of four Klebsiella pneumoniae and one Citrobacter freundii. The prevalence of faecal carriage of ESBL-producing organisms was 3.3%. Of these, 75% produced CTX-M-type enzymes followed by 22.7% SHV-producing strains. All faecal ESBL-producing strains were E. coli except for one Enterobacter cloacae and one Proteus mirabilis. This latter strain produced the PER-1 enzyme reported for the first time in Spain. CONCLUSIONS: The prevalence of ESBL-producing strains in stool samples was higher than that observed in clinical samples from the same period. The different types of ESBLs found were similar in both contexts. The most prevalent ESBLs were the CTX-M related enzymes, with nine different types, followed by SHV-12. PMID- 16244086 TI - Increasing incidence of quinolone resistance in human non-typhoid Salmonella enterica isolates in Korea and mechanisms involved in quinolone resistance. AB - OBJECTIVES: We investigated the trends of nalidixic acid resistance in human non typhoid Salmonella enterica in a Korean population, and examined some possible mechanisms involved in this resistance. METHODS: A total of 261 clinical strains were tested. For all strains, the MICs of nalidixic acid were determined. Nalidixic acid-resistant strains underwent further analysis, including determination of MICs of other antibiotics, mutation analysis within the topoisomerase genes, organic solvent tolerance test, western blotting for AcrA, marOR mutation analysis, ciprofloxacin accumulation test, and PCR for the qnr gene. The clonal relationships of Salmonella strains were examined by random amplified polymorphic DNA analysis. RESULTS: The incidence of nalidixic acid resistance increased from 1.8% in 1995-96 to 21.8% in 2000-02. The resistance rate was higher in S. enterica serotype Enteritidis (21.6%) than in serotype Typhimurium (12.1%). The nalidixic acid resistance rates in Salmonella Enteritidis varied according to the phage type (PT) and Salmonella Enteritidis PT 1 was most commonly associated with resistance to nalidixic acid. Several cases of clonal spread, especially by Salmonella Enteritidis PT 1, were identified. Of the 46 nalidixic acid-resistant strains, 43 had single mutations in the gyrA gene. Four strains were organic solvent-tolerant and were associated with decreased ciprofloxacin accumulation; three of these showed increased expression of AcrA and had novel mutations in marOR (84L). The qnr gene was not identified. CONCLUSIONS: Recently, the rate of nalidixic acid resistance in Korean clinical Salmonella strains markedly increased and it was partly due to the clonal spread of Salmonella Enteritidis, especially PT 1. The main mechanism of nalidixic acid resistance was a mutation in the gyrA region. PMID- 16244087 TI - Cost effectiveness of inhaled steroid withdrawal in outpatients with chronic obstructive pulmonary disease. AB - BACKGROUND: The evidence for the effectiveness and safety of inhaled corticosteroids (ICS) in chronic obstructive pulmonary disease (COPD) is inconclusive. This study determined the cost effectiveness of withdrawing fluticasone propionate (FP) in outpatients with COPD. METHODS: The cost effectiveness analysis was based on a randomised, placebo controlled FP withdrawal study. After a 4 month run in period on FP, patients were randomly assigned to continue FP 500 microg twice daily or to receive placebo for 6 months. A decision analytical model evaluated the 6 month incremental cost effectiveness of the ICS versus ICS withdrawal strategy. One way sensitivity analyses and a Monte Carlo simulation were performed to evaluate the robustness of the findings. RESULTS: The average patient with COPD in the FP group generated 511 in direct medical costs, including 238 for FP. The cost of the placebo strategy was 456. The higher direct drug cost of 212 per patient for the FP strategy during the 6 month follow up period compared with the placebo group was partially offset by a lower exacerbation and hospital admission cost of 157. The 6 month incremental cost effectiveness of the FP strategy compared with placebo was 110 per exacerbation prevented and 1286 per hospital admission prevented. CONCLUSIONS: Over a 6 month period, withdrawing FP in a pre-selected trial population of COPD patients led to absolute cost savings but with a higher rate of exacerbations and hospital admissions. PMID- 16244088 TI - Inhalation of stable dust extract prevents allergen induced airway inflammation and hyperresponsiveness. AB - BACKGROUND: Recent epidemiological studies have shown that growing up on a traditional farm provides protection from the development of allergic disorders such as hay fever and allergic asthma. We present experimental evidence that substances providing protection from the development of allergic diseases can be extracted from dust collected in stables of animal farms. METHODS: Stable dust was collected from 30 randomly selected farms located in rural regions of the Alps (Austria, Germany and Switzerland). The dust was homogenised with glass beads and extracted with physiological sodium chloride solution. This extract was used to modulate immune response in a well established mouse model of allergic asthma. RESULTS: Treatment of mice by inhalation of stable dust extract during sensitisation to ovalbumin inhibited the development of airway hyperresponsiveness and airway eosinophilia upon challenge, as well as the production of interleukin 5 by splenocytes and of antigen specific IgG(1) and IgE. Dust extract also suppressed the generation of human dendritic cells in vitro. The biological activity of the dust extract was not exclusively mediated by lipopolysaccharide. CONCLUSIONS: Stable dust from animal farms contains strong immune modulating substances. These substances can interfere with the development of both cellular and humoral immunity against allergens, thus suppressing allergen sensitisation, airway inflammation, and airway hyperresponsiveness in a murine model of allergic asthma. PMID- 16244089 TI - Progression of lung disease on computed tomography and pulmonary function tests in children and adults with cystic fibrosis. AB - BACKGROUND: A study was undertaken to compare the ability of computed tomographic (CT) scores and pulmonary function tests to detect changes in lung disease in children and adults with cystic fibrosis (CF). METHODS: CT scans and pulmonary function tests were retrospectively studied in a cohort of patients with CF aged 5-52 years for whom two or three CT scans at 3 year intervals were available, together with pulmonary function test results. All CT scans were scored by two observers. Pulmonary function results were expressed as percentage predicted and Z scores. RESULTS: Of 119 patients studied, two CT scans were available in 92 patients and three in 24. CT (composite and component) scores and lung function both deteriorated significantly (p<0.02). Peripheral bronchiectasis worsened by 1.7% per year in children (p<0.0001) and by 1.5% per year in adults (p<0.0001). Bronchiectasis worsened in 68 of 92 patients while forced expiratory volume in 1 second (FEV1) worsened in 54 of 92 patients; bronchiectasis also deteriorated in 27 patients with stable or improving FEV1. The CT score (and its components) and pulmonary function tests showed similar rates of deterioration in adults and children (p>0.09). CONCLUSION: The peripheral bronchiectasis CT score deteriorates faster and more frequently than lung function parameters in children and adults with CF, which indicates that pulmonary function tests and CT scans measure different aspects of CF lung disease. Our data support previous findings that the peripheral bronchiectasis CT score has an added value to pulmonary function tests in monitoring CF lung disease. PMID- 16244091 TI - Comparison of shuttle walk with measured peak oxygen consumption in patients with operable lung cancer. AB - BACKGROUND: The relationship between the shuttle walk test and peak oxygen consumption in patients with lung cancer has not previously been reported. A study was undertaken to examine this relationship in patients referred for lung cancer surgery to test the hypothesis that the shuttle walk test would be useful in this clinical setting. METHODS: 125 consecutive patients with potentially operable lung cancer were prospectively recruited. Each performed same day shuttle walking and treadmill walking tests. RESULTS: Shuttle walk distances ranged from 104 m to 1020 m and peak oxygen consumption ranged from 9 to 35 ml/kg/min. The shuttle walk distance significantly correlated with peak oxygen consumption (r = 0.67, p<0.001). All 55 patients who achieved more than 400 m on the shuttle test had a peak oxygen consumption of at least 15 ml/kg/min. Seventy of 125 patients failed to achieve 400 m on the shuttle walk test; in 22 of these the peak oxygen consumption was less than 15 ml/kg/min. Nine of 17 patients who achieved less than 250 m had a peak oxygen consumption of more than 15 ml/kg/min. CONCLUSION: The shuttle walk is a useful exercise test to assess potentially operable lung cancer patients with borderline lung function. However, it tends to underestimate exercise capacity at the lower range compared with peak oxygen consumption. Our data suggest that patients achieving 400 m on the shuttle walk test do not require formal measurement of oxygen consumption. In patients failing to achieve this distance we recommend assessment of peak oxygen consumption, particularly in those unable to walk 250 m, because a considerable proportion would still qualify for surgery as they had an acceptable peak oxygen consumption. PMID- 16244090 TI - Differences in radiological/HRCT findings in eosinophilic bronchitis and asthma: implication for bronchial responsiveness. AB - BACKGROUND: Airway hyperresponsiveness in asthmatics is considered to be one of the major consequences of airway inflammation and remodelling. Airway responsiveness is normal in patients with eosinophilic bronchitis (EB), despite eosinophilic inflammation of the airways comparable to that which occurs in asthmatics. Comparisons between asthma and EB should clarify the changes in airway morphology that are related specifically to AHR in asthmatics. METHODS: Eighteen asthmatic patients, 15 patients with EB, and 11 healthy subjects were recruited. Airway wall area percentage (WA%), centrilobular prominence, and air trapping were compared using thin slice section computed tomography. RESULTS: WA% was significantly greater in asthmatics than in patients with EB (72 (3.1)% v 54 (2.1)%, p = 0.032) and was similar in EB patients and controls (54 (2.1)% v 57 (1.8)%, p>0.05). Centrilobular prominence and air trapping were similar in EB patients and asthmatics and were significantly greater than in controls. CONCLUSION: WA% rather than air trapping or centrilobular prominence may be associated with the airway hyperresponsiveness that occurs in asthmatics but not in patients with EB. PMID- 16244092 TI - Diet and asthma in Dutch school children (ISAAC-2). AB - BACKGROUND: The rise in the prevalence of asthma in western societies may be related to changed dietary habits. Epidemiological studies in children have shown inverse associations of asthma related outcomes with intake of fruits, vegetables, dairy and whole grain products, and fish. In contrast to most previous studies, we used both questionnaire and clinical data to define asthma. METHODS: Intake of the abovementioned foods was studied in relation to asthma in 598 Dutch children aged 8-13 years. Dietary intake was estimated using a parent completed semi-quantitative food frequency questionnaire. Current wheeze and current asthma were defined based on questionnaire data. More complex end points were defined using information on bronchial hyperresponsiveness (BHR) and atopic sensitisation as well. Linear associations were studied using logistic regression analysis and odds ratios presented for the highest versus the lowest tertile of intake. In the final models, adjustments were made for maternal educational level, foreign descent, and total energy intake. RESULTS: The intake of whole grain products and of fish was inversely associated with asthma. Adjusted odds ratios for the independent associations with whole grains and fish were 0.46 (95% CI 0.19 to 1.10) and 0.34 (95% CI 0.13 to 0.85) for current asthma and 0.28 (95% CI 0.08 to 0.99) and 0.12 (95% CI 0.02 to 0.66) for atopic asthma with BHR. Similar results were observed for current wheeze and atopic wheeze with BHR. Intake of (citrus) fruits, vegetables, and dairy products showed no clear associations with asthma end points. CONCLUSIONS: Our findings suggest that a high intake of whole grain products and fish may have a protective effect against asthma in children. PMID- 16244094 TI - Exhaled nitric oxide in sarcoidosis. AB - BACKGROUND: Increased production of nitric oxide (NO) by the lower respiratory tract is viewed as a marker of airway inflammation in asthma and bronchiectasis. NO is a potentially important immune modulator, inhibiting the release of several key pro-inflammatory cytokines. As sarcoidosis is characterised by granulomatous airway inflammation, we hypothesised that exhaled NO levels might be raised in sarcoidosis and correlate with the morphological extent and functional severity of disease. METHODS: Fifty two patients with sarcoidosis (29 men) of mean age 42 years underwent thin section computed tomography (CT), pulmonary function tests, and measurement of exhaled NO. RESULTS: Exhaled NO levels (median 6.8 ppb, range 2.4-21.8) did not differ significantly from values in 44 control subjects, and were not related to the extent of individual CT abnormalities or the level of pulmonary function impairment. CONCLUSION: Exhaled NO levels are not increased in pulmonary sarcoidosis. PMID- 16244096 TI - Physics must join with biology in better assessing risk from low-dose irradiation. AB - This review summarises the complex response of mammalian cells and tissues to low doses of ionising radiation. This thesis encompasses induction of DNA damage, and adaptive protection against both renewed damage and against propagation of damage from the basic level of biological organisation to the clinical expression of detriment. The induction of DNA damage at low radiation doses apparently is proportional to absorbed dose at the physical/chemical level. However, any propagation of such damage to higher levels of biological organisation inherently follows a sigmoid function. Moreover, low-dose-induced inhibition of damage propagation is not linear, but instead follows a dose-effect function typical for adaptive protection, after an initial rapid rise it disappears at doses higher than approximately 0.1-0.2 Gy to cells. The particular biological response duality at low radiation doses precludes the validity of the linear-no-threshold hypothesis in the attempt to relate absorbed dose to cancer. In fact, theory and observation support not only a lower cancer incidence than expected from the linear-no-threshold hypothesis, but also a reduction of spontaneously occurring cancer, a hormetic response, in the healthy individual. PMID- 16244095 TI - A multi-compartment model for slow bronchial clearance of insoluble particles- extension of the ICRP human respiratory tract models. AB - To incorporate the various mechanisms that are presently assumed to be responsible for the experimentally observed slow bronchial clearance into the HRTM, a multi-compartment model was developed to simulate the clearance of insoluble particles in the tracheobronchial tree of the human lung. The new model considers specific mass transfer paths that may play an important role for slow bronchial clearance. These include the accumulation of particulate mass in the periciliary sol layer, phagocytosis of stored particles by airway macrophages and uptake of deposited mass by epithelial cells. Besides the gel layer representing fast mucociliary clearance, all cellular and non-cellular units involved in the slow clearance process are described by respective compartments that are connected by specific transfer rates. The gastrointestinal tract and lymph nodes are included into the model as final accumulation compartments, to which mass is transferred via the airway route and the transepithelial path. Predicted retention curves correspond well with previously published data. PMID- 16244093 TI - Hormone replacement therapy, body mass index and asthma in perimenopausal women: a cross sectional survey. AB - BACKGROUND: Hormone replacement therapy (HRT) and obesity both appear to increase the risk of asthma. A study was undertaken to investigate the association of HRT with asthma and hay fever in a population of perimenopausal women, focusing on a possible interaction with body mass index (BMI). METHODS: A postal questionnaire was sent to population based samples in Denmark, Estonia, Iceland, Norway, and Sweden in 1999-2001, and 8588 women aged 25-54 years responded (77%). Pregnant women, women using oral contraceptives, and women <46 years were excluded. Analyses included 2206 women aged 46-54 years of which 884 were menopausal and 540 used HRT. Stratified analyses by BMI in tertiles were performed. RESULTS: HRT was associated with an increased risk for asthma (OR 1.57 (95% CI 1.07 to 2.30)), wheeze (OR 1.60 (95% CI 1.22 to 2.10)), and hay fever (OR 1.48 (95% CI 1.15 to 1.90)). The associations with asthma and wheeze were significantly stronger among women with BMI in the lower tertile (asthma OR 2.41 (95% CI 1.21 to 4.77); wheeze OR 2.04 (95% CI 1.23 to 3.36)) than in heavier women (asthma: p(interaction) = 0.030; wheeze: p(interaction) = 0.042). Increasing BMI was associated with more asthma (OR 1.08 (95% CI 1.05 to 1.12) per kg/m2). This effect was only found in women not taking HRT (OR 1.10 (95% CI 1.05 to 1.14) per kg/m2); no such association was detected in HRT users (OR 1.00 (95% CI 0.92 to 1.08) per kg/m2) (p(interaction) = 0.046). Menopause was not significantly associated with asthma, wheeze, or hay fever. CONCLUSIONS: In perimenopausal women there is an interaction between HRT and BMI in the effects on asthma. Lean women who were HRT users had as high a risk for asthma as overweight women not taking HRT. It is suggested that HRT and overweight increase the risk of asthma through partly common pathways. PMID- 16244097 TI - Activity Concentration of 137Cs in soil samples from Punjab province (Pakistan) and estimation of gamma-ray dose rate for external exposure. AB - Punjab is one of the most populated provinces of Pakistan having 34 districts. There is always a need to have baseline background level information about 137Cs and the corresponding gamma-ray doses to the population. In this respect, soil samples were collected to a depth of 30-50 cm from all 34 district headquarters of the Punjab province of Pakistan and 137Cs activity concentrations were measured. 137Cs activity concentration was found to be in the range from 1.1 +/- 1.0 to 5.3 +/- 2.5 Bq kg(-1). The average value of estimated external gamma-ray dose rate from all 34 districts of Punjab province was computed to be 0.18 +/- 0.07 nSv h(-1) from 137Cs in soil samples. The measured 137Cs activity concentration range was compared with the reported ranges in the literature from some of the other locations in the world. Results obtained in this study show that 137Cs concentration is of a lower level in the investigated area. The average value of estimated external effective dose rate is found far below the dose rate limit of 1.0 mSv y(-1) for members of the general public recommended by the International Commission on Radiological Protection (ICRP) as well as the external gamma radiation dose of 0.48 mSv y(-1) received per head from the natural sources of radiation assessed by UNSCEAR (2000). It is concluded that 137Cs soil contamination does not pose radiation hazards to the population in the investigated areas. PMID- 16244098 TI - The European Radiobiology Archives (ERA)--content, structure and use illustrated by an example. AB - The European Radiobiology Archives (ERA), supported by the European Commission and the European Late Effect Project Group (EULEP), together with the US National Radiobiology Archives (NRA) and the Japanese Radiobiology Archives (JRA) have collected all information still available on long-term animal experiments, including some selected human studies. The archives consist of a database in Microsoft Access, a website, databases of references and information on the use of the database. At present, the archives contain a description of the exposure conditions, animal strains, etc. from approximately 350,000 individuals; data on survival and pathology are available from approximately 200,000 individuals. Care has been taken to render pathological diagnoses compatible among different studies and to allow the lumping of pathological diagnoses into more general classes. 'Forms' in Access with an underlying computer code facilitate the use of the database. This paper describes the structure and content of the archives and illustrates an example for a possible analysis of such data. PMID- 16244099 TI - Effects of a psychosocial intervention on breast self-examination attitudes and behaviors. AB - An educational intervention to promote breast self-examinations (BSEs) among young women was tested. In a group (intervention versus control) x time (Session 1 versus Session 2) mixed design, 172 college females were randomly assigned to either an intervention or control condition. Both groups attended two sessions; the second session was 48 hours after the first. The intervention consisted of an essay, lecture, video portraying young survivors of breast cancer, group discussions, self-test and instructions on performing BSEs. The control group had the same format; however, the information was focused on nutrition and exercise. Participants in the intervention group scored higher on rational problem solving and behavioral intentions, suggesting that the intervention increased adaptive responses to breast cancer threat. Conversely, control participants scored significantly higher on maladaptive reactions (e.g. hopelessness, avoidance and fatalistic religiosity) to breast cancer threat. For intervention participants, the initial decline in maladaptive reactions remained stable at 3-month follow up, but adaptive reactions decreased. Intervention participants had greater confidence in performing BSEs compared with controls but performed them on an irregular basis. Results were interpreted in terms of protection motivation theory, a model that applies the social psychology of persuasion to preventive health. PMID- 16244100 TI - Economic evaluation of interventions for problem drinking and alcohol dependence: do within-family external effects make a difference? AB - AIMS: To propose methods for the inclusion of within-family external effects in clinical and economic evaluations. To demonstrate the extent of bias due to the exclusion of within-family external effects when measuring the relative performance of interventions for problem drinking and alcohol dependence. METHODS: The timing and magnitude of treatment effects are modified to accommodate the external health-related quality of life impact of having a problem or dependent drinker in the family home. RESULTS: The inclusion of within family external effects reduces cost per QALY estimates of interventions for problem drinking and alcohol dependence thereby improving the performance of all evaluated interventions. In addition, the inclusion of within-family external effects improves the relative performance of interventions targeted at those with moderate-to-severe alcohol dependence as compared to interventions targeted at less severe alcohol problems. CONCLUSIONS: Failure to take account of external effects in clinical and economic evaluations results in an uneven playing field. Interventions with readily quantifiable health benefits (where social costs and benefits are predominantly comprised of private costs and benefits) are at a distinct advantage when competing for public funding against interventions with quantitatively important external effects. PMID- 16244101 TI - Unravelling the mystery of work-related upper limb disorder. PMID- 16244103 TI - Secondary Cushing's syndrome in children with juvenile idiopathic arthritis following intra-articular triamcinolone acetonide administration. PMID- 16244102 TI - Cerebral MRI is necessary in patients with rheumatoid arthritis and uveitis before undergoing therapy with TNF-alpha blocking agents. PMID- 16244104 TI - Ceramide catabolism critically controls survival of human dendritic cells. AB - The regulation of dendritic cell (DC) survival is crucial for the modulation of adaptive immunity. Ceramide is a lipid mediator of the stress response, which accumulates intracellularly during DC differentiation. We found that ceramide levels are tightly regulated in human DCs and that the pharmacological inhibition of enzymes responsible for ceramide catabolism, such as ceramidases and sphingosine kinases, sensitizes DCs to ceramide-induced cell death. It is important that inhibition of sphingosine kinases, during lipopolysaccharide stimulation, causes extensive ceramide accumulation and death of DCs. These data indicate that ceramide catabolism regulates survival of human DCs and reveal novel potential targets for the pharmacological manipulation of the immune response. PMID- 16244105 TI - Interferon-activated neutrophils store a TNF-related apoptosis-inducing ligand (TRAIL/Apo-2 ligand) intracellular pool that is readily mobilizable following exposure to proinflammatory mediators. AB - Neutrophils are versatile cells, which play a role, not only in inflammatory processes but also in immune and antitumoral responses. Recently, we have reported that interferon (IFN)-activated neutrophils are able to release biologically active tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL/APO2 ligand), a molecule exerting selective, apoptotic activities toward tumor and virus-infected cells, as well as immunoregulatory functions on activated T lymphocytes. Herein, we show that only a minor fraction of the total TRAIL, newly synthesized by IFN-activated neutrophils within 24 h, is released outside, the rest being retained intracellularly, mainly in secretory vesicles and light membrane fractions. We demonstrate that the intracellular pool of TRAIL present in IFN-pretreated neutrophils is rapidly mobilizable to the cell surface and can be secreted following exposure to proinflammatory mediators such as TNF alpha, lipopolysaccharide, formyl-methionyl-leucyl-phenylalanine, CXC chemokine ligand 8/interleukin-8, insoluble immunocomplexes, and heat shock protein Gp96. These various proinflammatory agonists functioned as effective secretagogue molecules only, in that they failed to augment TRAIL mRNA expression or TRAIL de novo synthesis in freshly isolated neutrophils or cultured with or without IFN. In addition, supernatants from IFN-treated neutrophils stimulated with proinflammatory mediators induced the apoptosis of target cells more effectively than supernatants from neutrophils activated with IFNs alone. Collectively, our results uncover a novel mechanism, whereby the release of soluble TRAIL by neutrophils can be greatly amplified and further reinforce the notion that neutrophils are important cells in tumor surveillance and immunomodulation. PMID- 16244106 TI - Abnormal regulation of the cytoskeletal regulator Rho typifies macrophages of the major murine models of spontaneous autoimmunity. AB - Macrophages (mphi) from prediseased mice of all the major murine models of spontaneous autoimmunity have an identical defect in cytokine expression that is triggered by serum and/or apoptotic cells. We show here that mphi from prediseased mice of the same models of spontaneous autoimmunity share a serum dependent defect in the activity of Rho, a cytoplasmic G protein and cytoskeletal regulator. Affected strains include those developing lupus (BXSB, LG, MRL/l+, MRL/lpr, NZBWF1) and autoimmune diabetes (nonobese diabetic). No similar defect in Rho activity occurred in seven control strains. In the presence of serum, Rho activity in mphi from all autoimmune-prone strains was reduced to less than 10% of that in control mice. In contrast, under serum-free conditions, Rho activity was completely normal in autoimmune-prone mphi. The activities of Ras, another cytoplasmic G protein, and Rac and Cdc42, two additional G protein regulators of the cytoskeleton, were regulated normally in autoimmune-prone strains. Serum dependent dysregulation of Rho was associated with multiple abnormalities, including increased adhesion to various surfaces, a more spread dendritic morphology, and an altered actin cytoskeletal organization. Our results suggest that mphi from multiple, genetically diverse, autoimmune-prone strains share a mutation or allelic difference affecting signal transduction within a specific Rho-regulatory pathway. PMID- 16244107 TI - Induction of intracellular calcium elevation by Delta9-tetrahydrocannabinol in T cells involves TRPC1 channels. AB - We have reported previously that Delta9-tetrahydrocannabinol (Delta9-THC) treatment of resting human and murine splenic T cells robustly elevated intracellular calcium ([Ca2+]i). The objective of the present investigation was to examine the putative role of [Ca2+]i store depletion and store-operated calcium (SOC) and receptor-operated cation (ROC) channels in the mechanism by which Delta9-THC increases [Ca2+]i in the cannabinoid-2 receptor-expressing human peripheral blood-acute lymphoid leukemia (HPB-ALL) human T cell line. By using the smooth endoplasmic reticulum Ca2+-ATPase pump inhibitor, thapsigargin, and the ryanodine receptor antagonist, 8-bromo-cyclic adenosine diphosphate ribose, we demonstrate that the Delta9-THC-mediated elevation in [Ca2+]i occurs independently of [Ca2+]i store depletion. Furthermore, the ROC channel inhibitor, SK&F 96365 was more efficacious at attenuating the Delta9-THC-mediated elevation in [Ca2+]i than SOC channel inhibitors, 2-aminoethoxydiphenyl borate and La3+. Recently, several members of the transient receptor potential canonical (TRPC) channel subfamily have been suggested to operate as SOC or ROC channels. In the present studies, treatment of HPB-ALL cells with 1-oleoyl-2-acetyl-sn-glycerol (OAG), a cell-permeant analog of diacylglycerol (DAG), which gates several members of the TRPC channel subfamily, rapidly elevated [Ca2+]i, as well as prevented a subsequent, additive elevation in [Ca2+]i by Delta9-THC, independent of protein kinase C. Reverse transcriptase-polymerase chain reaction analysis for TRPC1-7 showed that HPB-ALL cells express detectable mRNA levels of only TRPC1. Finally, small interference RNA knockdown of TRPC1 attenuated the Delta9-THC mediated elevation of [Ca2+]i. Collectively, these results suggest that Delta9 THC-induced elevation in [Ca2+]i is attributable entirely to extracellular calcium influx, which is independent of [Ca2+]i store depletion, and is mediated, at least partially, through the DAG-sensitive TRPC1 channels. PMID- 16244108 TI - Latent TGF-beta1-transduced CD4+ T cells suppress the progression of allergic encephalomyelitis. AB - Systemic injection of small amounts of transforming growth factor-beta (TGF beta), a cytokine produced by lymphoid and other cells, has a profound effect in protecting mice from the inflammatory demyelinating lesions of experimental allergic encephalomyelitis (EAE; an animal model for multiple sclerosis). However, TGF-beta has side-effects, which might be avoided if the cells producing TGF-beta can be delivered to the affected site in the nervous system to insure its local release in small amounts. Myelin basic protein (MBP)-specific, cloned CD4+ T cells were engineered by retroviral transduction to produce latent TGF beta. Studies about the spontaneous form of EAE in T cell receptor (TCR) transgenic recombination-activating gene (RAG)-1(-/-) mice showed that essentially all of the MBP-specific, TCR-transgenic RAG-1(-/-) (BALB/cxB10.PL)F1 mice develop spontaneous EAE by the age of 11 weeks. By 12 weeks, 25-50% of the mice have died from disease. A single injection of TGF-beta1-transduced T helper cell type 1 (Th1) cells significantly protected the mice from EAE, and untransduced Th1 cells did not protect. MBP-specific BALB/c Th2 clones, transduced with TGF-beta1-internal ribosome entry site-green fluorescent protein (GFP) significantly reduced EAE induction by untransduced Th1 cells in RAG-1(-/-) B10.PL mice. Furthermore, the GFP+ TGF-beta1-producing Th2 cells were detectable in the spinal cords of the injected mice. PMID- 16244109 TI - The interferon-inducible gene, Ifi204, is transcriptionally activated in response to M-CSF, and its expression favors macrophage differentiation in myeloid progenitor cells. AB - The interferon-inducible (Ifi)204 gene was isolated as a macrophage-colony stimulating factor (M-CSF)-responsive gene using a gene trap approach in the myeloid interleukin-3 (IL-3)-dependent FD-Fms cell line, which differentiates in macrophages in response to M-CSF. Here, we show that Ifi204 was transcriptionally activated in response to M-CSF, and FD-Fms cells decreased their growth and committed toward a macrophage morphology; this induction was abrogated when the differentiation signal of the M-CSF receptor was blocked; the Ifi204 gene was also induced during macrophage differentiation controlled by leukemia inhibitory factor; and the Ifi204 gene is expressed in different mature monocyte/macrophage cells. Finally, we showed that enforced expression of Ifi204 strongly decreased IL-3- and M-CSF-dependent proliferation and conversely, favored macrophage differentiation of FD-Fms cells in response to M-CSF. Altogether, these results demonstrate that the Ifi204 gene is activated during macrophage development and suggest that the Ifi204 protein may act as a regulator of the balance between proliferation and differentiation. Moreover, this study suggests that other members of the Ifi family might act as regulators of hematopoiesis under the control of hemopoietic cytokines. PMID- 16244110 TI - Common methodology is inadequate for studies on the microbicidal activity of neutrophils. AB - Microbicidal activity of neutrophils is usually measured by colony-counting techniques after cell lysis in distilled water. While studying the effect of the reduced nicotinamide adenine dinucleotide phosphate-oxidase inhibitor diphenyleneiodonium (DPI) on the staphylocidal activity of neutrophils, we obtained inconsistent results: various degrees of inhibition in some experiments and no effect in others. The lysis step, i.e., dilution of neutrophils in distilled water, was the source of error. Cell-associated microorganisms were not dispersed effectively by this treatment. We overcame this problem by using water at pH 11 for cell lysis. Under these conditions, killing was inhibited completely and reproducibly by DPI. Here, we show that cell lysis in distilled water is incomplete and leads to an overestimate of microbial killing. This hinders identification of partial defects and makes complete defects appear as partial. We found that DPI-treated neutrophils and chronic granulomatous disease neutrophils were completely defective in killing of Staphylococcus aureus and Candida albicans and partially defective in killing of Escherichia coli after lysis with water pH 11, whereas after lysis in distilled water, killing of S. aureus and C. albicans was approximately 60% and approximately 70% of control killing, respectively, and killing of E. coli was normal. Likewise, killing of S. aureus by myeloperoxidase-deficient neutrophils was severely impaired after lysis in water pH 11 but appeared normal after lysis in distilled water. As most studies about neutrophil microbicidal activity have been performed using distilled water, our findings indicate that previous data about killing defects and the effects of agents that modulate microbicidal activity of neutrophils should be re-evaluated. PMID- 16244111 TI - Activation of human eosinophils via P2 receptors: novel findings and future perspectives. AB - A growing body of information indicates that release of intracellular nucleotides represents an important way to modulate several cell pathways in physiological or pathological conditions. Nucleotides released as a consequence of cell damage, cell stress, bacterial infection, or other noxious stimuli signal at a class of plasma membrane receptors--P2 receptors--activating diverse intracellular pathways in many tissues and organs. For example, nucleotides secreted in the airway system control chloride/liquid secretion, goblet cell degranulation, and ciliary beat frequency. Several studies indicate that nucleotides play a role in airway diseases through their action on multiple cell types, including mast cells, dendritic cells, neurons, and eosinophils. Recent work by us and other groups led to the identification and characterization of P2 receptors expressed by human eosinophils. In this review, we will summarize recent developments in this field and put forward a hypothesis about the role of P2 receptors in pathophysiological conditions where eosinophils are major players. PMID- 16244112 TI - Synergistic enhancement of cytokine-induced human monocyte matrix metalloproteinase-1 by C-reactive protein and oxidized LDL through differential regulation of monocyte chemotactic protein-1 and prostaglandin E2. AB - C-reactive protein (CRP) and oxidized LDL (ox-LDL) are associated with inflammatory lesions, such as coronary artery disease, in which monocytes and matrix metalloproteinases (MMPs) may play a major role in the rupture of atherosclerotic plaques. Monocytes are recruited to inflammation sites by monocyte chemoattractant protein-1 (MCP-1), which may also participate in the activation of monocytes. The objective of this study was to compare the individual and combined effect of CRP and ox-LDL on human monocyte MMP-1 and the role of MCP-1 in this effect. Although CRP or ox-LDL failed to induce MMP-1 in control monocytes, these molecules enhanced MMP-1 production induced by tumor necrosis factor alpha (TNF-alpha) and granulocyte macrophage-colony stimulating factor (GM-CSF) with a synergistic increase in MMP-1 occurring in the presence of both mediators. Enhancement of MMP-1 by CRP and ox-LDL was attributable to a differential increase in MCP-1 and prostaglandin E2(PGE2). CRP, at physiological concentrations, induced high levels of MCP-1 and relatively low levels of PGE2, whereas ox-LDL caused a significant enhancement of PGE2 with little affect on MCP 1. Accordingly, CRP- and ox-LDL-induced MMP-1 production by monocytes was inhibited by anti-MCP-1 antibodies and indomethacin, respectively. Moreover, addition of exogenous MCP-1 or PGE2 enhanced MMP-1 production by TNF-alpha- and GM-CSF-stimulated monocytes. These results show that the combination of CRP and ox-LDL can cause a synergistic enhancement of the role of monocytes in inflammation, first, by increasing MCP-1, which attracts more monocytes and directly enhances MMP-1 production by activated monocytes, and second, by elevating PGE2 production, which also leads to higher levels of MMP-1. PMID- 16244113 TI - Endotoxin tolerance induces selective alterations in neutrophil function. AB - Endotoxin tolerance has the potential to limit phagocyte responses to Toll-like receptor (TLR) agonists, but the role of tolerance in regulating neutrophil responses is unknown. We investigated neutrophil responses to prolonged lipopolysaccharide (LPS) exposure and observed induction of tolerance in intracellular signaling pathways and respiratory burst. These effects were not prevented by granulocyte macrophage-colony stimulating factor (GM-CSF) pretreatment, and tolerized neutrophils retained the ability to respond to GM-CSF and other survival factors with a delay in apoptosis. In addition, LPS-exposed neutrophils showed continued generation of CXC chemokine ligand 8, which was not reduced in tolerized cells. Induction of tolerance was associated with a loss of TLR4 surface expression. Tolerance, therefore, induces a selective reprogramming of neutrophil function, but cells retain a predominantly proinflammatory phenotype. PMID- 16244114 TI - Cytokine-regulated expression and inhibitory function of FcgammaRIIB1 and -B2 receptors in human dendritic cells. AB - Dendritic cells (DC) capture immune complexes (IC) via Fc receptors for immunoglobulin G FcgammaRII and elicit antigen presentation and protective antitumoral immune response in mice. Two protocols are commonly used to differentiate human monocyte-derived DC in vitro. They associate granulocyte macrophage-colony stimulating factor (CM-CSF) with interleukin (IL)-4 or IL-13. In this study, we first assessed the ability of the two types of DC to initiate an immune response against an IC-linked antigen. We evidenced that IL-4 and IL-13 DC display comparable lymphocyte stimulatory capacity and similar lifetimes. We next characterized FcgammaRIIs expressed by pure populations of circulating myeloid DC (BDCA1+DC), IL-4, and IL-13 DC. We highlighted the expression of FcgammaRIIA, -B1, and -B2 by pure populations of BDCA1 myeloid DCs and IL-4 and IL-13 DC. Moreover, IL-4 and IL-13 DC displayed greater FcgammaRIIB expression than monocytes but a comparable FcgammaRIIA. We next investigated the FcgammaRIIB mechanism of action. We evidenced that deleting FcgammaRIIB increased the ability of IC-pulsed DC to stimulate autologous lymphocytes. FcgammaRIIB acted by lowering IC uptake, surface expression of costimulation molecules, and cytokine release. Finally, the balance between activating FcgammaRIIA/inhibitory FcgammaRIIB (B1+B2) could be modulated in vitro by inflammation mediators. By lowering FcgammaRIIB expression without significantly affecting FcgammaRIIA, prostaglandin E2 (PGE-2) appeared to be a major regulator of this balance. IL 1beta and tumor necrosis factor alpha were also found to potentiate PGE-2 action. Altogether, our results evidence an inhibitory role for FcgammaRIIB in human DC and provide an easy way to possibly improve in vitro the induction of immune response against IC-linked antigen. PMID- 16244115 TI - Modulation of phenotype and function of dendritic cells by a therapeutic synthetic killer peptide. AB - The strong microbicidal effects of an engineered synthetic killer peptide (KP), which functionally mimics a fungal killer toxin, have been demonstrated extensively. Beta-glucan has been identified as a receptor for KP on fungal cell walls. Although the direct microbicidal and related therapeutic effects have been studied in depth, no information currently exists about the interaction of KP with immune cells. In this study, we exploited the possibility of KP binding to different murine immune cell populations. The results demonstrate that KP binds selectively to dendritic cells (DC) and to a lesser extent, to macrophages but not to lymphocytes and neutrophils; KP binding possibly occurs through major histocompatibility complex (MHC) class II, CD16/32, and cellular molecules recognized by anti-specific intercellular adhesion molecule-grabbing nonintegrin R1 antibodies; and KP modulates the expression of costimulatory and MHC molecules on DC and improves their capacity to induce lymphocyte proliferation. These findings provide evidence that this synthetic KP interacts selectively with DC and modulating their multiple functions, might also serve to improve the immune antimicrobial response. PMID- 16244116 TI - PAF-receptor antagonists, lovastatin, and the PTK inhibitor genistein inhibit H2O2 secretion by macrophages cultured on oxidized-LDL matrices. AB - Adhesion of mononuclear phagocytes (Macs) to extracellular matrices containing oxidized low-density lipoproteins (oxLDL) stimulates these cells to secrete reactive oxygen species (e.g., O2-, H2O2) that are believed to promote atherogenesis. Current in vitro systems designed to measure Mac H2O2 secretion in response to oxLDL show that these cells secrete H2O2 for only a few hours after plating. The slow onset and chronicity of atherogenesis, however, suggested to us that Mac ROS secretion might be sustained for much longer periods when Macs are maintained in an environment resembling that in the intima of arteries undergoing atherogenesis. The findings reported here confirm this suggestion. They show that Macs maintained on collagen IV matrices containing oxLDL in medium containing human plasma-derived serum secrete H2O2 continuously and in large amounts for at least 11 days. Using this system we tested the effects of compounds known to attenuate atherogenesis in vivo. Platelet-activating factor (PAF) receptor antagonists, lovastatin, and the isoflavone protein tyrosine kinase (PTK) inhibitor genistein each reduced H2O2 secretion by Macs maintained on oxLDL containing matrices by approximately 60%. Lovastatin's inhibitory effect was blocked completely by addition of geranylgeranyl pyrophosphate to the medium. We conclude that matrix-bound and oxidized lipoproteins stimulate Macs to produce H2O2 continuously and in large quantities via a pathway that involves PAF receptors and PTK and is reversibly blocked by inhibitors of protein prenylation. PMID- 16244117 TI - Age trends and age norms for the NEO Personality Inventory-3 in adolescents and adults. AB - The NEO Personality Inventory-3 (NEO-PI-3) is a modification of the Revised NEO Personality Inventory (NEO-PI-R) designed to be more understandable to adolescents. Data from adults aged 21 to 91 showed that the NEO-PI-3 also functions as well or better than the NEO-PI-R in adults. Age trends from combined adolescent (n = 500) and adult (n = 635) samples confirmed previous cross sectional findings and demonstrated the importance of studying age changes especially at the facet level and during the decade of the 20s. Normative data for self-report and observer rating forms for adolescents, younger and older adults, and all adults are discussed, as well as for a combined-age group. It is argued that combined-age norms may be most appropriate for depicting the personality scores of individuals, but the utility for some purposes of within age group scores is also acknowledged. PMID- 16244118 TI - Using the short mood and feelings questionnaire to detect depression in detained adolescents. AB - The Mood and Feelings Questionnaire (MFQ) is examined for its utility in screening youth in juvenile justice settings for depression. In a cross-sectional study conducted at King County Juvenile Detention Center, a representative sample of 228 detained adolescents complete structured assessments, including the MFQ and the Massachusetts Youth Screening Instrument. Fifty youth also complete the Voice-Diagnostic Interview Schedule for Children. Internal reliability coefficient for the MFQ short form (SMFQ) is = .87. Factor analysis produces a unifactorial scale with item loadings of .43 to .78. At SMFQ cutoff > or = 10, sensitivity and specificity are optimized at 1.00/0.79. Prevalence of major depressive disorder is estimated at 32.1% (95% Confidence Interval = 25.3% to 39.2%). The SMFQ shows potential for depression screening of detained adolescents. PMID- 16244120 TI - Paper and voice MAYSI-2: format comparability and concordance with the voice DISC IV. AB - The authors examine the comparability of paper and voice formats of the Massachusetts Youth Screening Instrument-Second Version (MAYSI-2) as well as each format's concordance with the Voice Diagnostic Interview Schedule for Children (DISC) among adjudicated youth. Comparability is assessed among 248 youths admitted to a South Carolina Assessment Center. Mean scores and alpha coefficients are calculated, and area under the curve and positive and negative predictive values are used to compare concordance to the DISC. Paper and voice formats are significantly correlated, have similar alpha coefficients, and have comparable concordance with the DISC, suggesting that the MAYSI-2 paper and voice formats are comparable instruments. The voice format may be preferable to the paper format for screening justice youth for mental health problems because it may reduce incomplete data and increase reporting of stigmatized behaviors and because of its ability to automatically generate scored reports and aggregate data. PMID- 16244119 TI - Use of the adolescent SASSI in a juvenile correctional setting. AB - The Substance Abuse Subtle Screening Inventory-Adolescent (SASSI-A) is used in evaluation and treatment planning for incarcerated juveniles. Validity of the SASSI-A in a juvenile correctional facility was examined using archival data. Findings generally support the validity of SASSI-A substance use scales. However, there is concern regarding the potential for ethnic bias in this setting. Cut scores suggest that the SASSI-A may best be used for detecting problematic alcohol consumption using the Face Valid Alcohol Scale > or = 3. Future studies should more closely investigate whether the three underlying dimensions of the SASSI-A are useful in treatment planning. Results are presented in light of the relatively new SASSI-A2. PMID- 16244121 TI - A simplified technique for scoring DSM-IV personality disorders with the Five Factor Model. AB - The current study compares the use of two alternative methodologies for using the Five-Factor Model (FFM) to assess personality disorders (PDs). Across two clinical samples, a technique using the simple sum of selected FFM facets is compared with a previously used prototype matching technique. The results demonstrate that the more easily calculated counts perform as well as the similarity scores that are generated by the prototype matching technique. Optimal diagnostic thresholds for the FFM PD counts are computed for identifying patients who meet diagnostic criteria for a specific PD. These threshold scores demonstrate good sensitivity in receiver operating characteristics analyses, suggesting their usefulness for screening purposes. Given the ease of this scoring procedure, the FFM count technique has obvious clinical utility. PMID- 16244122 TI - Construct validity of the Balloon Analog Risk Task (BART): associations with psychopathy and impulsivity. AB - To continue research assessing the validity of the Balloon Analog Risk Task (BART) as a measure of risk taking, the BART was administered to a nonforensic sample of individuals with varying levels of psychopathic characteristics. Construct validity of the BART was evaluated by measuring risk-taking behavior in relation to self-reported psychopathy, anxiety, psychophysiological data, and demographic variables. Supporting the construct validity of the BART, while controlling for variance accounted for by interrelated variables in the context of hierarchical regression analyses, higher self-reported psychopathy was significantly predictive of increased risk taking on the BART. Findings also revealed interesting gender differences in self-reported psychopathy and a trend for men to engage in riskier behavior. Limitations and research directions are presented. PMID- 16244123 TI - WAIS digit span-based indicators of malingered neurocognitive dysfunction: classification accuracy in traumatic brain injury. AB - The present study determined specificity and sensitivity to malingered neurocognitive dysfunction (MND) in traumatic brain injury (TBI) for several Wechsler Adult Intelligence Scale (WAIS) Digit Span scores. TBI patients (n = 344) were categorized into one of five groups: no incentive, incentive only, suspect, probable MND, and definite MND. Performance of 1,063 nonincentive patients (e.g., cerebrovascular accident, memory disorder) was also examined. Digit Span scores included reliable digit span, maximum span forward both trials correct, maximum span forward, combined maximum forward and backward span, Digit Span scaled score, maximum span backward both trials correct, and maximum span backward. In TBI, sensitivity to MND ranged from 15% to greater than 30% at specificities of 92% to 98%. Patient groups with documented brain pathology had higher false-positive error rates. These results replicate previous known-groups malingering studies and provide valuable data supporting the WAIS Digit Span scores in detection and diagnosis of malingering. PMID- 16244124 TI - Graduate student WAIS-III scoring accuracy is a function of full scale IQ and complexity of examiner tasks. AB - Research on the Wechsler Adult Intelligence Scale-Revised and Wechsler Adult Intelligence Scale-Third Edition (WAIS-III) suggests that practicing clinical psychologists and graduate students make item-level scoring errors that affect IQ, index, and subtest scores. Studies have been limited in that Full-Scale IQ (FSIQ) and examiner administration, recording, and scoring tasks have not been systematically varied. In this study, graduate student participants score a high (FSIQ = 112) and low (FSIQ = 85) IQ record form in one of two stimulus conditions: digitized film clips (N = 13) or partially completed record forms (N = 11). Results demonstrate that examiners are less accurate in the high IQ condition, and that recording examinee responses from scoring video clips results in more scoring errors. Obtained FSIQs are significantly higher than criterion IQ scores in the high IQ condition (8.46 for video condition, 2.55 for record form condition). Self-reported proficiency in WAIS-III administration and scoring is positively related to number of scoring errors. PMID- 16244125 TI - A comparison of two mental status examinations in an inpatient psychiatric sample. AB - This study compares the psychometric properties and clinical use of the Mini Mental Status Exam (MMSE) and the Modified MMSE (3MS) in patients admitted to an acute medical-psychiatric inpatient unit. Internal consistencies were .56 for the estimated MMSE and .72 for the 3MS. Regression analyses revealed that the 3MS was a significant predictor of both length of hospital stay (LOS) and the need for additional services postdischarge. Regressions were also used to evaluate the contribution of the four new items contained in the 3MS: Word Generation, Similarities, Second Recall, and Date and Place of Birth. The 3MS was a significant predictor of LOS and the need for discharge services, whereas the MMSE did not contribute to any regression model. Word Generation was found to be a significant predictor of patients' needing posthospital services. Results support prior findings of the 3MS's superiority over the (estimated) MMSE and help to demonstrate the continued importance of cognitive screening in psychiatric samples. PMID- 16244129 TI - Homology modeling revealed more than 20,000 rRNA internal transcribed spacer 2 (ITS2) secondary structures. AB - Structural genomics meets phylogenetics and vice versa: Knowing rRNA secondary structures is a prerequisite for constructing rRNA alignments for inferring phylogenies, and inferring phylogenies is a precondition to understand the evolution of such rRNA secondary structures. Here, both scientific worlds go together. The rRNA internal transcribed spacer 2 (ITS2) region is a widely used phylogenetic marker. Because of its high variability at the sequence level, correct alignments have to take into account structural information. In this study, we examine the extent of the conservation in structure. We present (1) the homology modeled secondary structure of more than 20,000 ITS2 covering about 14,000 species; (2) a computational approach for homology modeling of rRNA structures, which additionally can be applied to other RNA families; and (3) a database providing about 25,000 ITS2 sequences with their associated secondary structures, a refined ITS2 specific general time reversible (GTR) substitution model, and a scoring matrix, available at http://its2.bioapps.biozentrum.uni wuerzburg.de. PMID- 16244128 TI - Binding of misacylated tRNAs to the ribosomal A site. AB - To test whether the ribosome displays specificity for the esterified amino acid and the tRNA body of an aminoacyl-tRNA (aa-tRNA), the stabilities of 4 correctly acylated and 12 misacylated tRNAs in the ribosomal A site were determined. By introducing the GAC (valine) anticodon into each tRNA, a constant anticodon.codon interaction was maintained, thus removing concern that different anticodon.codon strengths might affect the binding of the different aa-tRNAs to the A site. Surprisingly, all 16 aa-tRNAs displayed similar dissociation rate constants from the A site. These results suggest that either the ribosome is not specific for different amino acids and tRNA bodies when intact aa-tRNAs are used or the specificity for the amino acid side chain and tRNA body is masked by a conformational change upon aa-tRNA release. PMID- 16244130 TI - Ribosomes containing mutants of L4 ribosomal protein from Thermus thermophilus display multiple defects in ribosomal functions and sensitivity against erythromycin. AB - Protein L4 from Thermus thermophilus (TthL4) was heterologously overproduced in Escherichia coli cells. To study the implication of the extended loop of TthL4 in the exit-tunnel and peptidyltransferase functions, the highly conserved E56 was replaced by D or Q, while the semiconserved G55 was changed to E or S. Moreover, the sequence -G55E56- was inverted to -E55G56-. When we incorporated these mutants into E. coli ribosomes and investigated their impact on poly(Phe) synthesis, high variations in the synthetic activity and response to erythromycin of the resulting ribosomes were observed. In the absence of erythromycin, ribosomes harboring mutations G55E and E56D in TthL4 protein were characterized by low activity in synthesizing poly(Phe) and decreased capability in binding tRNA at the A site. On the other hand, ribosomes possessing mutations G55E, G55S, G55E-E56G, or E56Q in TthL4 protein were unexpectedly more sensitive to erythromycin. Evidence in support of these findings was drawn by in vivo experiments, assessing the erythromycin sensitivity of E. coli cells expressing wild-type or mutant TthL4 proteins. Our results emphasize the role of the extended loop of L4 ribosomal protein in the exit-tunnel and peptidyltransferase center functions. PMID- 16244131 TI - The Kluyveromyces lactis gamma-toxin targets tRNA anticodons. AB - Kluyveromyces lactis killer strains secrete a heterotrimeric toxin (zymocin), which causes an irreversible growth arrest of sensitive yeast cells. Despite many efforts, the target(s) of the cytotoxic gamma-subunit of zymocin has remained elusive. Here we show that three tRNA species tRNA(Glu)(mcm(5)s(2)UUC), tRNA(Lys)(mcm(5)s(2)UUU), and tRNA(Gln)(mcm(5)s(2)UUG) are the targets of gamma toxin. The toxin inhibits growth by cleaving these tRNAs at the 3' side of the modified wobble nucleoside 5-methoxycarbonylmethyl-2-thiouridine (mcm(5)s(2)U). Transfer RNA lacking a part of or the entire mcm(5) group is inefficiently cleaved by gamma-toxin, explaining the gamma-toxin resistance of the modification deficient trm9, elp1-elp6, and kti11-kti13 mutants. The K. lactis gamma-toxin is the first eukaryotic toxin shown to target tRNA. PMID- 16244132 TI - Yeast Puf3 mutants reveal the complexity of Puf-RNA binding and identify a loop required for regulation of mRNA decay. AB - The eukaryotic Puf proteins regulate mRNA translation and degradation by binding the 3' untranslated regions of target mRNAs. Crystal structure analysis of a human Puf bound to RNA suggested a modular mode of binding, with specific amino acids within each of eight repeat domains contacting a single nucleotide of the target RNA. Here we study the mechanism by which the yeast Puf3p binds and stimulates the degradation of COX17 mRNA. Mutation of the predicted RNA-binding positions of Puf3p to those found in Puf5p demonstrated that a single amino acid change in Puf3p abolished detectable binding to COX17. Since this amino acid position in both Puf3p and Puf5p is predicted to contact an adenine in the respective target RNAs, the amino acid in Puf3p must play a more critical role in promoting COX17 interaction. In contrast, an amino acid change in the third repeat of Puf3p, which interacts with the only divergent nucleotide between the Puf3p and Puf5p targets, had no effect on binding COX17. These results argue that a simple set of rules cannot reliably link specific amino acid positions with target specificity. Each of these amino acid changes in Puf3p enhanced binding to the Puf5p target HO RNA, suggesting a different mode of binding to this target. Finally, we identified an outer surface loop that was dispensable for binding but was required to promote both rapid deadenylation and subsequent decapping of the COX17 mRNA, most likely as a point of protein-protein interactions. PMID- 16244133 TI - An artificial riboswitch for controlling pre-mRNA splicing. AB - Riboswitches, as previously reported, are natural RNA aptamers that regulate the expression of numerous bacterial metabolic genes in response to small molecule ligands. It has recently been shown that these RNA genetic elements are also present near the splice site junctions of plant and fungal introns, thus raising the possibility of their involvement in regulating mRNA splicing. Here it is shown for the first time that a riboswitch can be engineered to regulate pre-mRNA splicing in vitro. We show that insertion of a high-affinity theophylline binding aptamer into the 3' splice site (3' ss) region of a model pre-mRNA (AdML Theo29AG) enables its splicing to be repressed by the addition theophylline. Our results indicate that the location of 3' ss AG within the aptamer plays a crucial role in conferring theophylline-dependent control of pre-mRNA splicing. We also show that theophylline-mediated control of pre-mRNA splicing is highly specific by first demonstrating that a small molecule ligand similar in shape and size to theophylline had no effect on the splicing of AdML-Theo29AG pre-mRNA. Second, theophylline failed to exert any influence on the splicing of a pre-mRNA that does not contain its binding site. Third, theophylline specifically blocks the step II of the splicing reaction. Finally, we provide evidence that theophylline dependent control of pre-mRNA splicing is functionally relevant. PMID- 16244134 TI - Solution probing of metal ion binding by helix 27 from Escherichia coli 16S rRNA. AB - Helix (H)27 from Escherichia coli 16S ribosomal (r)RNA is centrally located within the small (30S) ribosomal subunit, immediately adjacent to the decoding center. Bacterial 30S subunit crystal structures depicting Mg(2+) binding sites resolve two magnesium ions within the vicinity of H27: one in the major groove of the G886-U911 wobble pair, and one within the GCAA tetraloop. Binding of such metal cations is generally thought to be crucial for RNA folding and function. To ask how metal ion-RNA interactions in crystals compare with those in solution, we have characterized, using solution NMR spectroscopy, Tb(3+) footprinting and time resolved fluorescence resonance energy transfer (tr-FRET), location, and modes of metal ion binding in an isolated H27. NMR and Tb(3+) footprinting data indicate that solution secondary structure and Mg(2+) binding are generally consistent with the ribosomal crystal structures. However, our analyses also suggest that H27 is dynamic in solution and that metal ions localize within the narrow major groove formed by the juxtaposition of the loop E motif with the tandem G894-U905 and G895-U904 wobble pairs. In addition, tr-FRET studies provide evidence that Mg(2+) uptake by the H27 construct results in a global lengthening of the helix. We propose that only a subset of H27-metal ion interactions has been captured in the crystal structures of the 30S ribosomal subunit, and that small-scale structural dynamics afforded by solution conditions may contribute to these differences. Our studies thus highlight an example for differences between RNA metal ion interactions observed in solution and in crystals. PMID- 16244135 TI - Simple, quantitative primer-extension PCR assay for direct monitoring of microRNAs and short-interfering RNAs. AB - There has been a surge of interest in the biology of microRNAs and the technology of RNA interference. We describe a simple, robust, inexpensive assay for quantitative analysis of microRNAs and short-interfering RNAs. The method relies on primer extension conversion of RNA to cDNA by reverse transcription followed by quantitative, real-time PCR. Technical parameters critical to the success of the assay are presented. Measurements of microRNA levels are sensitive, with most assays allowing measurements in the femtomolar range, which corresponds to tens of copies per cell or less. The assay has a high dynamic range and provides linear readout over differences in microRNA concentrations that span 6-7 orders of magnitude. The assay is capable of discriminating between related microRNA family members that differ by subtle sequence differences. We used the method for quantitative analysis of six microRNAs across 12 tissue samples. The data confirm striking variation in the patterns of expression of these noncoding regulatory RNAs. PMID- 16244136 TI - Three maize leaf ferredoxin:NADPH oxidoreductases vary in subchloroplast location, expression, and interaction with ferredoxin. AB - In higher plants, ferredoxin (Fd):NADPH oxidoreductase (FNR) catalyzes reduction of NADP+ in the final step of linear photosynthetic electron transport and is also implicated in cyclic electron flow. We have identified three leaf FNR isoenzymes (LFNR1, LFNR2, and LFNR3) in maize (Zea mays) chloroplasts at approximately equivalent concentrations. Fractionation of chloroplasts showed that, while LFNR3 is an exclusively soluble enzyme, LFNR1 is only found at the thylakoid membrane and LFNR2 has a dual location. LFNR1 and LFNR2 were found to associate with the cytochrome b6f complex following its partial purification. We cloned LFNR3 and produced all three isoenzymes as stable, soluble proteins. Measurement of Fd reduction ability showed no significant differences between these recombinant enzymes. Column chromatography revealed variation between the interaction mechanisms of LFNR1 and LFNR2 with Fd, as detected by differential dependence on specific intermolecular salt bridges and variable sensitivity of interactions to changes in pH. A comparison of LFNR transcripts in leaves of plants grown on variable nitrogen regimes revealed that LFNR1 and LFNR2 transcripts are relatively more abundant under conditions of high demand for NADPH. These results are discussed in terms of the functional differentiation of maize LFNR isoenzymes. PMID- 16244137 TI - Identification of three urease accessory proteins that are required for urease activation in Arabidopsis. AB - Urease is a nickel-containing urea hydrolase involved in nitrogen recycling from ureide, purine, and arginine catabolism in plants. The process of urease activation by incorporation of nickel into the active site is a prime example of chaperone-mediated metal transfer to an enzyme. Four urease accessory proteins are required for activation in Klebsiella aerogenes. In plants urease accessory proteins have so far been only partially defined. Using reverse genetic tools we identified four genes that are necessary for urease activity in Arabidopsis (Arabidopsis thaliana; ecotypes Columbia and Nossen). Plants bearing T-DNA or Ds element insertions in either the structural gene for urease or in any of the three putative urease accessory genes AtureD, AtureF, and AtureG lacked the corresponding mRNAs and were defective in urease activity. In contrast to wild type plants, the mutant lines were not able to support growth with urea as the sole nitrogen source. To investigate whether the identified accessory proteins would be sufficient to support eukaryotic urease activation, the corresponding cDNAs were introduced into urease-negative Escherichia coli. In these bacteria, urease activity was observed only when all three plant accessory genes were coexpressed together with the plant urease gene. Remarkably, plant urease activation occurred as well in cell-free E. coli extracts, but only in extracts from cells that had expressed all three accessory proteins. The future molecular dissection of the plant urease activation process may therefore be performed in vitro, providing a powerful tool to further our understanding of the biochemistry of chaperone-mediated metal transfer processes in plants. PMID- 16244138 TI - Enhanced tolerance to environmental stress in transgenic plants expressing the transcriptional coactivator multiprotein bridging factor 1c. AB - Abiotic stresses cause extensive losses to agricultural production worldwide. Acclimation of plants to abiotic conditions such as drought, salinity, or heat is mediated by a complex network of transcription factors and other regulatory genes that control multiple defense enzymes, proteins, and pathways. Associated with the activity of different transcription factors are transcriptional coactivators that enhance their binding to the basal transcription machinery. Although the importance of stress-response transcription factors was demonstrated in transgenic plants, little is known about the function of transcriptional coactivators associated with abiotic stresses. Here, we report that constitutive expression of the stress-response transcriptional coactivator multiprotein bridging factor 1c (MBF1c) in Arabidopsis (Arabidopsis thaliana) enhances the tolerance of transgenic plants to bacterial infection, heat, and osmotic stress. Moreover, the enhanced tolerance of transgenic plants to osmotic and heat stress was maintained even when these two stresses were combined. The expression of MBF1c in transgenic plants augmented the accumulation of a number of defense transcripts in response to heat stress. Transcriptome profiling and inhibitor studies suggest that MBF1c expression enhances the tolerance of transgenic plants to heat and osmotic stress by partially activating, or perturbing, the ethylene response signal transduction pathway. Present findings suggest that MBF1 proteins could be used to enhance the tolerance of plants to different abiotic stresses. PMID- 16244139 TI - Functional interaction between two transcription factors involved in the developmental regulation of a small heat stress protein gene promoter. AB - Hahsp17.6G1 is the promoter of a small heat stress protein (sHSP) from sunflower (Helianthus annuus) that is activated during zygotic embryogenesis, but which does not respond to heat stress. We report here the cloning of a transcription factor (TF), sunflower drought-responsive element binding factor 2 (HaDREB2), by one-hybrid interaction with functional cis-elements in Hahsp17.6G1. We have analyzed the functional interaction between HaDREB2 and a second transcription factor, sunflower heat stress factor A9 (HaHSFA9), which was previously assigned to the regulation of Hahsp17.6G1. HaDREB2 and HaHSFA9 synergistically trans activate the Hahsp17.6G1 promoter in bombarded sunflower embryos. This synergistic interaction is heat stress factor (HSF) specific and requires the binding of both factors to the promoter. The C-terminal region of HaHSFA9 is sufficient for the HSF specificity. Our results represent an example of a functional interaction between members of the Apetala 2 (HaDREB2) and HSF (HaHSFA9) families of transcription factors. We suggest new roles in zygotic embryogenesis for specific members of the AP2 transcription factor family. PMID- 16244140 TI - Differential expression of sucrose-phosphate synthase isoenzymes in tobacco reflects their functional specialization during dark-governed starch mobilization in source leaves. AB - Sucrose (Suc)-phosphate synthase (SPS) plays a crucial role in the synthesis of Suc in photosynthetic and nonphotosynthetic tissues. Several isoforms of SPS exist in dicotyledonous plants that can be grouped into the different families A, B, and C. To explore whether functional differences between the SPS gene families might exist, we characterized a representative for each family from tobacco (Nicotiana tabacum). RNA-blot analysis revealed a distinct expression pattern for each of the three SPS genes. While the A-family member (NtSPSA) was found to be expressed in all tissues examined, expression of the B isoform (NtSPSB) was mainly confined to the reproductive organs and NtSPSC mRNA was exclusively detected in mature source leaves. We used RNA interference to assess the in planta function of NtSPSA and C. While silencing of NtSPSA had no detectable influence on leaf carbohydrate metabolism, reduction of NtSPSC led to an increase in leaf starch content by a factor of 3 to 8. Further analysis revealed that starch accumulation in NtSPSC-silenced plants was not due to an increased partitioning of carbon into starch, but rather showed that starch mobilization was impaired. The transgenic plants were unable to efficiently mobilize their transitory leaf starch during a prolonged period of darkness and accumulated maltose as a major intermediate of starch breakdown. NtSPSC mRNA level increased appreciably during the dark period while transcript levels of the other isoforms showed no diurnal changes. Together, these results suggest that NtSPSC is specifically involved in the synthesis of Suc during starch mobilization in the dark. The roles of the other SPS isoforms are discussed. PMID- 16244141 TI - Suppression of allene oxide cyclase in hairy roots of Medicago truncatula reduces jasmonate levels and the degree of mycorrhization with Glomus intraradices. AB - During the symbiotic interaction between Medicago truncatula and the arbuscular mycorrhizal (AM) fungus Glomus intraradices, an endogenous increase in jasmonic acid (JA) occurs. Two full-length cDNAs coding for the JA-biosynthetic enzyme allene oxide cyclase (AOC) from M. truncatula, designated as MtAOC1 and MtAOC2, were cloned and characterized. The AOC protein was localized in plastids and found to occur constitutively in all vascular tissues of M. truncatula. In leaves and roots, MtAOCs are expressed upon JA application. Enhanced expression was also observed during mycorrhization with G. intraradices. A partial suppression of MtAOC expression was achieved in roots following transformation with Agrobacterium rhizogenes harboring the MtAOC1 cDNA in the antisense direction under control of the cauliflower mosaic virus 35S promoter. In comparison to samples transformed with 35SuidA, roots with suppressed MtAOC1 expression exhibited lower JA levels and a remarkable delay in the process of colonization with G. intraradices. Both the mycorrhization rate, quantified by fungal rRNA, and the arbuscule formation, analyzed by the expression level of the AM-specific gene MtPT4, were affected. Staining of fungal material in roots with suppressed MtAOC1 revealed a decreased number of arbuscules, but these did not exhibit an altered structure. Our results indicate a crucial role for JA in the establishment of AM symbiosis. PMID- 16244142 TI - A mutation in the anticodon of a single tRNAala is sufficient to confer auxin resistance in Arabidopsis. AB - Auxin-resistant mutants have been useful for dissecting the mechanisms that underlie auxin-mediated biological processes. Here we report the isolation and molecular characterization of a novel auxin-resistant mutant in Arabidopsis (Arabidopsis thaliana). Like known mutated AUX/IAA transcription factors, the mutant described here displayed dominant resistance to exogenously supplied auxins (sirtinol, 2,4-dichlorophenoxyacetic acid, indole-3-acetic acid) and a host of pleiotropic phenotypes, including apical hook deformation, defects in lateral root development, reduced stature, and homozygous lethality. This mutant showed the same sensitivity to the ethylene precursor 1-aminocyclopropane carboxylic acid as wild-type plants, and retained the ability to induce IAA19 expression in response to exogenously supplied indole-3-acetic acid. To our surprise, these phenotypes were not caused by a mutation in an AUX/IAA gene, but rather a mutation in a tRNA(ala) gene in which the anticodon was found changed from CGC to CAC. Such a change results in a tRNA that is charged with alanine but recognizes the second most highly used valine codon in Arabidopsis. Therefore, the observed phenotypes are likely the composite of stochastic mutations of many proteins, including downstream effectors. PMID- 16244143 TI - Plant-derived transfer DNAs. AB - The transfer of DNA from Agrobacterium to plant cell nuclei is initiated by a cleavage reaction within the 25-bp right border of Ti plasmids. In an effort to develop all-native DNA transformation vectors, 50 putative right border alternatives were identified in both plant expressed sequence tags and genomic DNA. Efficacy tests in a tobacco (Nicotiana tabacum) model system demonstrated that 14 of these elements displayed at least 50% of the activity of conventional Agrobacterium transfer DNA borders. Four of the most effective plant-derived right border alternatives were found to be associated with intron-exon junctions. Additional elements were embedded within introns, exons, untranslated trailers, and intergenic DNA. Based on the identification of a single right border alternative in Arabidopsis and three in rice (Oryza sativa), the occurrence of this motif was estimated at a frequency of at least 0.8x10(-8). Modification of plasmid DNA sequences flanking the alternative borders demonstrated that both upstream and downstream sequences play an important role in initiating DNA transfer. Optimal DNA transfer required the elements to be preceded by pyrimidine residues interspaced by AC-rich trinucleotides. Alteration of this organization lowered transformation frequencies by 46% to 93%. Despite their weaker resemblance with left borders, right border alternatives also functioned effectively in terminating DNA transfer, if both associated with an upstream A[C/T]T[C/G]A[A/T]T[G/T][C/T][G/T][C/G]A[C/T][C/T][A/T] domain and tightly linked cytosine clusters at their junctions with downstream DNA. New insights in border region requirements were used to construct an all-native alfalfa (Medicago sativa) transfer DNA vector that can be used for the production of intragenic plants. PMID- 16244144 TI - Overexpression of AtCpNifS enhances selenium tolerance and accumulation in Arabidopsis. AB - Selenium (Se) is an essential element for many organisms but is toxic at higher levels. CpNifS is a chloroplastic NifS-like protein in Arabidopsis (Arabidopsis thaliana) that can catalyze the conversion of cysteine into alanine and elemental sulfur (S0) and of selenocysteine into alanine and elemental Se (Se0). We overexpressed CpNifS to investigate the effects on Se metabolism in plants. CpNifS overexpression significantly enhanced selenate tolerance (1.9-fold) and Se accumulation (2.2-fold). CpNifS overexpressors showed significantly reduced Se incorporation into protein, which may explain their higher Se tolerance. Also, sulfur accumulation was enhanced by approximately 30% in CpNifS overexpressors, both on media with and without selenate. Root transcriptome changes in response to selenate mimicked the effects observed under sulfur starvation. There were only a few transcriptome differences between CpNifS-overexpressing plants and wild type, besides the 25- to 40-fold increase in CpNifS levels. Judged from x ray analysis of near edge spectrum, both CpNifS overexpressors and wild type accumulated mostly selenate (Se(VI)). In conclusion, overexpression of this plant NifS-like protein had a pronounced effect on plant Se metabolism. The observed enhanced Se accumulation and tolerance of CpNifS overexpressors show promise for use in phytoremediation. PMID- 16244145 TI - PsbP protein, but not PsbQ protein, is essential for the regulation and stabilization of photosystem II in higher plants. AB - PsbP and PsbQ proteins are extrinsic subunits of photosystem II (PSII) and participate in the normal function of photosynthetic water oxidation. Both proteins exist in a broad range of the oxygenic photosynthetic organisms; however, their physiological roles in vivo have not been well defined in higher plants. In this study, we established and analyzed transgenic tobacco (Nicotiana tabacum) plants in which the levels of PsbP or PsbQ were severely down-regulated by the RNA interference technique. A plant that lacked PsbQ showed no specific phenotype compared to a wild-type plant. This suggests that PsbQ in higher plants is dispensable under the normal growth condition. On the other hand, a plant that lacked PsbP showed prominent phenotypes: drastic retardation of growth, pale green-colored leaves, and a marked decrease in the quantum yield of PSII evaluated by chlorophyll fluorescence. In PsbP-deficient plant, most PSII core subunits were accumulated in thylakoids, whereas PsbQ, which requires PsbP to bind PSII in vitro, was dramatically decreased. PSII without PsbP was hypersensitive to light and rapidly inactivated when the repair process of the damaged PSII was inhibited by chloramphenicol. Furthermore, thermoluminescence studies showed that the catalytic manganese cluster in PsbP-deficient leaves was markedly unstable and readily disassembled in the dark. The present results demonstrated that PsbP, but not PsbQ, is indispensable for the normal PSII function in higher plants in vivo. PMID- 16244146 TI - Genetic and molecular analyses of natural variation indicate CBF2 as a candidate gene for underlying a freezing tolerance quantitative trait locus in Arabidopsis. AB - Natural variation for freezing tolerance is a major component of adaptation and geographic distribution of plant species. However, little is known about the genes and molecular mechanisms that determine its naturally occurring diversity. We have analyzed the intraspecific freezing tolerance variation existent between two geographically distant accessions of Arabidopsis (Arabidopsis thaliana), Cape Verde Islands (Cvi) and Landsberg erecta (Ler). They differed in their freezing tolerance before and after cold acclimation, as well as in the cold acclimation response in relation to photoperiod conditions. Using a quantitative genetic approach, we found that freezing tolerance differences after cold acclimation were determined by seven quantitative trait loci (QTL), named FREEZING TOLERANCE QTL 1 (FTQ1) to FTQ7. FTQ4 was the QTL with the largest effect detected in two photoperiod conditions, while five other FTQ loci behaved as photoperiod dependent. FTQ4 colocated with the tandem repeated genes C-REPEAT BINDING FACTOR 1 (CBF1), CBF2, and CBF3, which encode transcriptional activators involved in the cold acclimation response. The low freezing tolerance of FTQ4-Cvi alleles was associated with a deletion of the promoter region of Cvi CBF2, and with low RNA expression of CBF2 and of several CBF target genes. Genetic complementation of FTQ4-Cvi plants with a CBF2-Ler transgene suggests that such CBF2 allelic variation is the cause of CBF2 misexpression and the molecular basis of FTQ4. PMID- 16244147 TI - Evidence that bicarbonate is not the substrate in photosynthetic oxygen evolution. AB - It is widely accepted that the oxygen produced by photosystem II of cyanobacteria, algae, and plants is derived from water. Earlier proposals that bicarbonate may serve as substrate or catalytic intermediate are almost forgotten, though not rigorously disproved. These latter proposals imply that CO2 is an intermediate product of oxygen production in addition to O2. In this work, we investigated this possible role of exchangeable HCO3- in oxygen evolution in two independent ways. (1) We studied a possible product inhibition of the electron transfer into the catalytic Mn4Ca complex during the oxygen-evolving reaction by greatly increasing the pressure of CO2. This was monitored by absorption transients in the near UV. We found that a 3,000-fold increase of the CO2 pressure over ambient conditions did not affect the UV transient, whereas the S(3) --> S(4) --> S(0) transition was half-inhibited by raising the O2 pressure only 10-fold over ambient, as previously established. (2) The flash-induced O2 and CO2 production by photosystem II was followed simultaneously with membrane inlet mass spectrometry under approximately 15% H2(18)O enrichment. Light flashes that revealed the known oscillatory O2 release failed to produce any oscillatory CO2 signal. Both types of results exclude that exchangeable bicarbonate is the substrate for (and CO2 an intermediate product of) oxygen evolution by photosynthesis. The possibility that a tightly bound carbonate or bicarbonate is a cofactor of photosynthetic water oxidation has remained. PMID- 16244148 TI - Salt stress in Thellungiella halophila activates Na+ transport mechanisms required for salinity tolerance. AB - Salinity is considered one of the major limiting factors for plant growth and agricultural productivity. We are using salt cress (Thellungiella halophila) to identify biochemical mechanisms that enable plants to grow in saline conditions. Under salt stress, the major site of Na+ accumulation occurred in old leaves, followed by young leaves and taproots, with the least accumulation occurring in lateral roots. Salt treatment increased both the H+ transport and hydrolytic activity of salt cress tonoplast (TP) and plasma membrane (PM) H(+)-ATPases from leaves and roots. TP Na(+)/H+ exchange was greatly stimulated by growth of the plants in NaCl, both in leaves and roots. Expression of the PM H(+)-ATPase isoform AHA3, the Na+ transporter HKT1, and the Na(+)/H+ exchanger SOS1 were examined in PMs isolated from control and salt-treated salt cress roots and leaves. An increased expression of SOS1, but no changes in levels of AHA3 and HKT1, was observed. NHX1 was only detected in PM fractions of roots, and a salt induced increase in protein expression was observed. Analysis of the levels of expression of vacuolar H(+)-translocating ATPase subunits showed no major changes in protein expression of subunits VHA-A or VHA-B with salt treatment; however, VHA-E showed an increased expression in leaf tissue, but not in roots, when the plants were treated with NaCl. Salt cress plants were able to distribute and store Na+ by a very strict control of ion movement across both the TP and PM. PMID- 16244149 TI - Ascorbic acid deficiency activates cell death and disease resistance responses in Arabidopsis. AB - Programmed cell death, developmental senescence, and responses to pathogens are linked through complex genetic controls that are influenced by redox regulation. Here we show that the Arabidopsis (Arabidopsis thaliana) low vitamin C mutants, vtc1 and vtc2, which have between 10% and 25% of wild-type ascorbic acid, exhibit microlesions, express pathogenesis-related (PR) proteins, and have enhanced basal resistance against infections caused by Pseudomonas syringae. The mutants have a delayed senescence phenotype with smaller leaf cells than the wild type at maturity. The vtc leaves have more glutathione than the wild type, with higher ratios of reduced glutathione to glutathione disulfide. Expression of green fluorescence protein (GFP) fused to the nonexpressor of PR protein 1 (GFP-NPR1) was used to detect the presence of NPR1 in the nuclei of transformed plants. Fluorescence was observed in the nuclei of 6- to 8-week-old GFP-NPR1 vtc1 plants, but not in the nuclei of transformed GFP-NPR1 wild-type plants at any developmental stage. The absence of senescence-associated gene 12 (SAG12) mRNA at the time when constitutive cell death and basal resistance were detected confirms that elaboration of innate immune responses in vtc plants does not result from activation of early senescence. Moreover, H2O2-sensitive genes are not induced at the time of systemic acquired resistance execution. These results demonstrate that ascorbic acid abundance modifies the threshold for activation of plant innate defense responses via redox mechanisms that are independent of the natural senescence program. PMID- 16244151 TI - Differential capacity for high-affinity manganese uptake contributes to differences between barley genotypes in tolerance to low manganese availability. AB - There is considerable variability among barley (Hordeum vulgare) genotypes in their ability to grow in soils containing a low level of plant available manganese (Mn). The physiological basis for the tolerance to low Mn availability is unknown. In this work, Mn2+ influx and compartmentation in roots of the Mn efficient genotype Vanessa and the Mn-inefficient genotype Antonia were investigated. Two separate Mn transport systems, mediating high-affinity Mn2+ influx at concentrations up to 130 nm and low-affinity Mn2+ influx at higher concentrations, were identified in both genotypes. The two genotypes differed only in high-affinity kinetics with the Mn-efficient genotype Vanessa having almost 4 times higher V(max) than the inefficient Antonia, but similar K(m) values. Online inductively coupled plasma-mass spectrometry measurements verified that the observed differences in high-affinity influx resulted in a higher Mn net uptake of Vanessa compared to Antonia. Further evidence for the importance of the differences in high-affinity uptake kinetics for Mn acquisition was obtained in a hydroponic system with mixed cultivation of the two genotypes at a continuously low Mn concentration (10-50 nm) similar to that occurring in soil solution. Under these conditions, Vanessa had a competitive advantage and contained 55% to 75% more Mn in the shoots than did Antonia. Subcellular compartmentation analysis of roots based on 54Mn2+ efflux established that up to 93% and 83% of all Mn was present in the vacuole in Vanessa and Antonia, respectively. It is concluded that differential capacity for high-affinity Mn influx contributes to differences between barley genotypes in Mn efficiency. PMID- 16244150 TI - Arabidopsis FHY1 protein stability is regulated by light via phytochrome A and 26S proteasome. AB - Phytochrome A (phyA) is the primary photoreceptor mediating responses to far-red light. Among the phyA downstream signaling components, Far-red Elongated Hypocotyl 1 (FHY1) is a genetically defined positive regulator of photomorphogenesis in far-red light. Both physiological and genomic characterization of the fhy1 mutants indicated a close functional relationship of FHY1 with phyA. Here, we showed that FHY1 is most abundant in young seedlings grown in darkness and is quickly down-regulated during further seedling development and by light exposure. By using light-insensitive 35S promoter-driven functional beta-glucuronidase-FHY1 and green fluorescent protein-FHY1 fusion proteins, we showed that this down-regulation of FHY1 protein abundance by light is largely at posttranscriptional level and most evident in the nuclei. The light triggered FHY1 protein reduction is primarily mediated through the 26S proteasome dependent protein degradation. Further, phyA is directly involved in mediating the light-triggered down-regulation of FHY1, and the dark accumulation of FHY1 requires functional pleiotropic Constitutive Photomorphogenic/De-Etiolated/Fusca proteins. Our data indicate that phyA, the 26S proteasome, and the Constitutive Photomorphogenic/De-Etiolated/Fusca proteins are all involved in the light regulation of FHY1 protein abundance during Arabidopsis (Arabidopsis thaliana) seedling development. PMID- 16244153 TI - The dynamic changes of tonoplasts in guard cells are important for stomatal movement in Vicia faba. AB - Stomatal movement is important for plants to exchange gas with environment. The regulation of stomatal movement allows optimizing photosynthesis and transpiration. Changes in vacuolar volume in guard cells are known to participate in this regulation. However, little has been known about the mechanism underlying the regulation of rapid changes in guard cell vacuolar volume. Here, we report that dynamic changes in the complex vacuolar membrane system play a role in the rapid changes of vacuolar volume in Vicia faba guard cells. The guard cells contained a great number of small vacuoles and various vacuolar membrane structures when stomata closed. The small vacuoles and complex membrane systems fused with each other or with the bigger vacuoles to generate large vacuoles during stomatal opening. Conversely, the large vacuoles split into smaller vacuoles and generated many complex membrane structures in the closing stomata. Vacuole fusion inhibitor, (2s,3s)-trans-epoxy-succinyl-l-leucylamido-3 methylbutane ethyl ester, inhibited stomatal opening significantly. Furthermore, an Arabidopsis (Arabidopsis thaliana) mutation of the SGR3 gene, which has a defect in vacuolar fusion, also led to retardation of stomatal opening. All these results suggest that the dynamic changes of the tonoplast are essential for enhancing stomatal movement. PMID- 16244152 TI - The polygalacturonase-inhibiting protein PGIP2 of Phaseolus vulgaris has evolved a mixed mode of inhibition of endopolygalacturonase PG1 of Botrytis cinerea. AB - Botrytis cinerea is a phytopathogenic fungus that causes gray mold in >1,000 plant species. During infection, it secretes several endopolygalacturonases (PGs) to degrade cell wall pectin, and among them, BcPG1 is constitutively expressed and is an important virulence factor. To counteract the action of PGs, plants express polygalacturonase-inhibiting proteins (PGIPs) that have been shown to inhibit a variety of PGs with different inhibition kinetics, both competitive and noncompetitive. The PG-PGIP interaction promotes the accumulation of oligogalacturonides, fragments of the plant cell wall that are general elicitors of plant defense responses. Here, we characterize the enzymatic activity of BcPG1 and investigate its interaction with PGIP isoform 2 from Phaseolus vulgaris (PvPGIP2) by means of inhibition assays, homology modeling, and molecular docking simulations. Our results indicate a mixed mode of inhibition. This is compatible with a model for the interaction where PvPGIP2 binds the N-terminal portion of BcPG1, partially covering its active site and decreasing the enzyme affinity for the substrate. The structural framework provided by the docking model is confirmed by site-directed mutagenesis of the residues that distinguish PvPGIP2 from the isoform PvPGIP1. The finding that PvPGIP2 inhibits BcPG1 with a mixed type kinetics further indicates the versatility of PGIPs to evolve different recognition specificities. PMID- 16244154 TI - Cryptochrome 1 contributes to blue-light sensing in pea. AB - Cryptochromes are widespread in higher plants but their physiological roles as blue-light photoreceptors have been examined in relatively few species. Screening in a phyA null mutant background has identified several blue-light response mutants in pea (Pisum sativum), including one that carries a substitution of a highly conserved glycine residue in the N-terminal photolyase-homologous domain of the pea CRY1 gene. Analyses of cry1, phyA, and phyB mutants show that all three photoreceptors contribute to seedling photomorphogenesis under high irradiance blue light, whereas phyA is the main photoreceptor active under low irradiances. Triple phyA phyB cry1 mutants grown under high-irradiance blue light are indistinguishable from dark-grown wild-type plants in length and leaf expansion but show a small residual response to higher-irradiance white light. Monogenic cry1 mutants have little discernable phenotype at the seedling stage, but later in development are more elongated than wild-type plants. In addition, the loss of cry1 moderates the short-internode phenotype of older phyA mutants, suggesting an antagonism between phyA and cry1 under some conditions. Pea cry1 has a small inhibitory effect on flowering under long and short days. However, the phyA cry1 double mutant retains a clear promotion of flowering in response to blue-light photoperiod extensions, indicating a role for one or more additional blue-light photoreceptors in the control of flowering in pea. PMID- 16244155 TI - Sec22 and Memb11 are v-SNAREs of the anterograde endoplasmic reticulum-Golgi pathway in tobacco leaf epidermal cells. AB - Distinct sets of soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptors (SNAREs) are distributed to specific intracellular compartments and catalyze membrane fusion events. Although the central role of these proteins in membrane fusion is established in nonplant systems, little is known about their role in the early secretory pathway of plant cells. Analysis of the Arabidopsis (Arabidopsis thaliana) genome reveals 54 genes encoding SNARE proteins, some of which are expected to be key regulators of membrane trafficking between the endoplasmic reticulum (ER) and the Golgi. To gain insights on the role of SNAREs of the early secretory pathway in plant cells, we have cloned the Arabidopsis v-SNAREs Sec22, Memb11, Bet11, and the t-SNARE Sed5, and analyzed their distribution in plant cells in vivo. By means of live cell imaging, we have determined that these SNAREs localize at the Golgi apparatus. In addition, Sec22 was also distributed at the ER. We have then focused on understanding the function of Sec22 and Memb11 in comparison to the other SNAREs. Overexpression of the v-SNAREs Sec22 and Memb11 but not of the other SNAREs induced collapse of Golgi membrane proteins into the ER, and the secretion of a soluble secretory marker was abrogated by all SNAREs. Our studies suggest that Sec22 and Memb11 are involved in anterograde protein trafficking at the ER-Golgi interface. PMID- 16244157 TI - Nectarin IV, a potent endoglucanase inhibitor secreted into the nectar of ornamental tobacco plants. Isolation, cloning, and characterization. AB - We have isolated and characterized the Nectarin IV (NEC4) protein that accumulates in the nectar of ornamental tobacco plants (Nicotiana langsdorffii x Nicotiana sanderae var LxS8). This 60-kD protein has a blocked N terminus. Three tryptic peptides of the protein were isolated and sequenced using tandem mass spectroscopy. These unique peptides were found to be similar to the xyloglucan specific fungal endoglucanase inhibitor protein (XEGIP) precursor in tomato (Lycopersicon esculentum) and its homolog in potato (Solanum tuberosum). A pair of oligonucleotide primers was designed based on the potato and tomato sequences that were used to clone a 1,018-bp internal piece of nec4 cDNA from a stage 6 nectary cDNA library. The remaining portions of the cDNA were subsequently captured by 5' and 3' rapid amplification of cDNA ends. Complete sequencing of the nec4 cDNA demonstrated that it belonged to a large family of homologous proteins from a wide variety of angiosperms. Related proteins include foliage proteins and seed storage proteins. Based upon conserved identity with the wheat (Triticum aestivum) xylanase inhibitor TAXI-1, we were able to develop a protein model that showed that NEC4 contains additional amino acid loops that are not found in TAXI-1 and that glycosylation sites are surface exposed. Both these loops and sites of glycosylation are on the opposite face of the NEC4 molecule from the site that interacts with fungal hemicellulases, as indicated by homology to TAXI-I. NEC4 also contains a region homologous to the TAXI-1 knottin domain; however, a deletion in this domain restructures the disulfide bridges of this domain, resulting in a pseudoknottin domain. Inhibition assays were performed to determine whether purified NEC4 was able to inhibit fungal endoglucanases and xylanases. These studies showed that NEC4 was a very effective inhibitor of a family GH12 xyloglucan-specific endoglucanase with a K(i) of 0.35 nm. However, no inhibitory activity was observed against other family GH10 or GH11 xylanases. The patterns of expression of the NEC4 protein indicate that, while expressed in nectar at anthesis, it is most strongly expressed in the nectary gland after fertilization, indicating that inhibition of fungal cell wall-degrading enzymes may be more important after fertilization than before. PMID- 16244159 TI - Does familiarity with the release site reduce the deflection induced by clock shifting? A comment to the paper by Gagliardo et al. (2005). PMID- 16244156 TI - Increased calcium levels and prolonged shelf life in tomatoes expressing Arabidopsis H+/Ca2+ transporters. AB - Here we demonstrate that fruit from tomato (Lycopersicon esculentum) plants expressing Arabidopsis (Arabidopsis thaliana) H(+)/cation exchangers (CAX) have more calcium (Ca2+) and prolonged shelf life when compared to controls. Previously, using the prototypical CAX1, it has been demonstrated that, in yeast (Saccharomyces cerevisiae) cells, CAX transporters are activated when the N terminal autoinhibitory region is deleted, to give an N-terminally truncated CAX (sCAX), or altered through specific manipulations. To continue to understand the diversity of CAX function, we used yeast assays to characterize the putative transport properties of CAX4 and N-terminal variants of CAX4. CAX4 variants can suppress the Ca2+ hypersensitive yeast phenotypes and also appear to be more specific Ca2+ transporters than sCAX1. We then compared the phenotypes of sCAX1- and CAX4-expressing tomato lines. The sCAX1-expressing tomato lines demonstrate increased vacuolar H(+)/Ca2+ transport, when measured in root tissue, elevated fruit Ca2+ level, and prolonged shelf life but have severe alterations in plant development and morphology, including increased incidence of blossom-end rot. The CAX4-expressing plants demonstrate more modest increases in Ca2+ levels and shelf life but no deleterious effects on plant growth. These findings suggest that CAX expression may fortify plants with Ca2+ and may serve as an alternative to the application of CaCl2 used to extend the shelf life of numerous agriculturally important commodities. However, judicious regulation of CAX transport is required to assure optimal plant growth. PMID- 16244161 TI - Path integration in a three-dimensional maze: ground distance estimation keeps desert ants Cataglyphis fortis on course. AB - In this study, we investigate the ability of desert ants to gauge the ground distances of sloped sections in a three-dimensional (3D) outbound path. Ground distance estimation, as opposed to a simple measurement of walking distances, is a necessary prerequisite for precise path integration in undulating terrain. We trained ants to visit a feeder along a path that included an angular turn as well as a 'hill', resulting in an outbound path with a distinct 3D structure. We then observed the ants' return path in a test field on level ground. From the angles of the ants' return path on the test field one can infer which property of the hill segment was fed into the ants' path integration module, the actual walking distance or the ground distance. The results show clearly that it is the ground distance that Cataglyphis fortis feeds into its path integrator, and suggest that the ants are able to keep an accurate home vector also in hilly terrain. PMID- 16244158 TI - Analysis of the cDNAs of hypothetical genes on Arabidopsis chromosome 2 reveals numerous transcript variants. AB - In the fully sequenced Arabidopsis (Arabidopsis thaliana) genome, many gene models are annotated as "hypothetical protein," whose gene structures are predicted solely by computer algorithms with no support from either expressed sequence matches from Arabidopsis, or nucleic acid or protein homologs from other species. In order to confirm their existence and predicted gene structures, a high-throughput method of rapid amplification of cDNA ends (RACE) was used to obtain their cDNA sequences from 11 cDNA populations. Primers from all of the 797 hypothetical genes on chromosome 2 were designed, and, through 5' and 3' RACE, clones from 506 genes were sequenced and cDNA sequences from 399 target genes were recovered. The cDNA sequences were obtained by assembling their 5' and 3' RACE polymerase chain reaction products. These sequences revealed that (1) the structures of 151 hypothetical genes were different from their predictions; (2) 116 hypothetical genes had alternatively spliced transcripts and 187 genes displayed polyadenylation sites; and (3) there were transcripts arising from both strands, from the strand opposite to that of the prediction and possible dicistronic transcripts. Promoters from five randomly chosen hypothetical genes (At2g02540, At2g31270, At2g33640, At2g35550, and At2g36340) were cloned into report constructs, and their expressions are tissue or development stage specific. Our results indicate at least 50% of hypothetical genes on chromosome 2 are expressed in the cDNA populations with about 38% of the gene structures differing from their predictions. Thus, by using this targeted approach, high throughput RACE, we revealed numerous transcripts including many uncharacterized variants from these hypothetical genes. PMID- 16244162 TI - Ontogenetic development of locomotion in small mammals--a kinematic study. AB - Comparative studies of locomotion indicate that limb design and performance are very similar in adult mammals of small to medium size. The present study was undertaken to test whether basic therian limb pattern is present during postnatal development. Kinematic data were collected from juveniles of two eutherian species in a cross-sectional study, using cinevideography. The tree shrew Tupaia glis and the cui Galea musteloides were selected because of their different reproductive strategies, which could result in differences in the development of locomotor abilities. The aims of this study were to describe the process by which young animals develop the adult pattern of locomotion and the extent to which this process varies in two species with very different postnatal ontogenies. Despite their different life histories, the development of kinematic parameters in the altricial tree shrew and the precocial cui are surprisingly similar. General limb design, performance, and timing of segment and joint movements in the young animals were similar to adults in both species, even from the first steps. Touch-down of the forelimb occurred at the position below the eye in all individuals and limb position was highly standardized at touch-down; no major changes in segment and joint angles were observed. Significant changes occurred at lift-off. With increasing body mass, limb segments rotated more caudally, which resulted in larger limb excursions and relatively longer steps. Developmental changes in locomotor abilities were similar in both species; only the time necessary to reach the adult performance was different. Despite the widely assumed maturity of locomotor abilities in precocial young, the first steps of the cui juveniles were not similar to the movements of adults. The adult locomotor pattern was reached within the first postnatal week in the cui and by the time they leave the nest in the tree shrew (39 days after birth; individual P39). These results suggest that during the evolution of precocial development only processes independent of exercise or gravity can be shifted into the intrauterine period. However, development of locomotor ability dependents on exercise, and adjustments and training occur during growth. Therefore, only the time necessary to reach maturity was clearly shortened in the precocial juvenile relative to the ancestral altricial condition. PMID- 16244163 TI - Oxygen delivery to the fish eye: root effect as crucial factor for elevated retinal PO2. AB - Although the retina has one of the highest metabolic rates among tissues, certain teleost fishes lack any vascular supply to this organ which, in combination with the overall thickness of the organ, results in extremely long diffusion distances. As the only way to compensate for these obstacles, oxygen partial pressure (PO2) in the eyes of such fish is elevated far above atmospheric values. Although not supported by any direct evidence, the enhancement of PO2 is considered to be related to the Root effect, the release upon acidification of Hb bound O2 into physical dissolution, possibly supported by counter-current multiplication similar to the loop of Henle. The present study evaluates the magnitude of intraocular PO2 enhancement under tightly controlled physiological conditions, to directly confirm the involvement of the Root effect on intraocular PO2 in the retina of rainbow trout Oncorhynchus mykiss. Intraocular PO2 was determined with special polarographic microelectrodes inserted into the eye. PO2 profiles established in vivo by driving electrodes through the entire retina yielded average PO2 values between 10 mmHg (1.3 kPa) at the inner retinal surface and 382 mmHg (50.9 kPa) close to the outer retinal limit (Bruch's membrane). According to estimates on the basis of the diffusion distances determined from sections of the retina (approximately 436 microm at the site of PO2 measurement) and literature data on specific oxygen consumption, the in vivo determined values would be sufficient to cover the oxygen demand of the retina with some safety margin. For a clear and direct in-tissue-test as to the involvement of the Root effect, an isolated in vitro eye preparation was established in order to avoid the problem of indirect blood supply to the eye from the dorsal aorta only via the pseudobranch, a hemibranch thought to modulate blood composition before entry of the eye. Any humoral effects (e.g. catecholamines) were eliminated by perfusing isolated eyes successively with standardized red blood cell (RBC) suspensions in Ringer, using trout (with Root) and human (lacking any Root effect) RBC suspension. To optimize perfusate conditions for maximal Root effect, the Root effect of trout RBCs was determined in vitro via graded acidification of individual samples equilibrated with standardized gas mixtures. During perfusion with trout RBC, PO2 at the outer retinal limit was 99 mmHg (13.2 kPa), but fell by a factor of 3.3 upon perfusion with human RBC in spite of higher total oxygen content (TO2 2.8 for trout vs 3.9 mmol l-1 for human RBC). Upon reperfusion with trout RBC, PO2 was restored immediately to the original value. This regularly observed pattern indicated a highly significant difference (P=0.003) between perfusion with trout (with Root effect; high retinal PO2) and perfusion with human (no Root effect; low retinal PO2) RBC suspension, thus clearly demonstrating that the Root effect is directly involved and a crucial prerequisite for the enhancement of PO2 in the retina of the teleost eye. PMID- 16244164 TI - Does swarming cause honey bees to update their solar ephemerides? AB - Spatial orientation in the social insects offers several examples of specialized learning mechanisms that underlie complex learning tasks. Here we study one of these systems: the processes by which honey bees update, or fail to update, their memories of the sun's daily pattern of movement (the solar ephemeris function) in relation to the landscape. Specifically, we ask whether bees that have initially learned the solar ephemeris function relative to a conspicuous treeline at their natal site can later realign the ephemeris to a differently oriented treeline. We first confirm and clarify an earlier finding that bees transplanted passively (by being carried) do not re-learn the solar ephemeris in relation to the new treeline. When they cannot detect the sun directly, as on overcast days, these transplanted bees use a solar ephemeris function appropriate for their natal site, despite days or weeks of experience at the new site. We then ask whether bees put through a swarming process as they are transplanted are induced to re learn the solar ephemeris function at the new site, as swarming is a natural process wherein bees transplant themselves. Most of the swarmed bees failed to re learn, even though they did extensive learning flights (in comparison with those of non-swarmed controls) as they first emerged from the hive at the new site. We hypothesize that the bees' representation of the solar ephemeris function is stored in an encapsulated cognitive module in which the ephemeris is inextricably linked to the reference landscape in which it was learned. PMID- 16244165 TI - Miniaturization (0.2 g) and evaluation of attachment techniques of telemetry transmitters. AB - We have developed a miniaturized very high frequency (VHF) transmitter design for radio-tagging small animals. The average mass of the circuitry is 0.084 g (range 0.081-0.087), hence, with the smallest power cell, complete tags weigh 0.2 g and have a life of 18-22 days. We further demonstrate that with such small tags the technique of attaching the device to the animal's body strongly affects the effective radiated power, and thus transmission ranges. Ideally the attachment couples the electronics to the animal's body as a ground plane for improved radiation. The transmitter allows the application of radio-tracking to be expanded to new taxa whose spatial behaviour and population dynamics are largely unknown, for example arthropods, reptiles or amphibians. The new design is also suitable for miniature implants and signal modulation with sensors. PMID- 16244167 TI - Cryoprotection by urea in a terrestrially hibernating frog. AB - The role of urea as a balancing osmolyte in osmotic adaptation is well known, but this 'waste product' also has myriad other functions in diverse taxa. We report that urea plays an important, previously undocumented role in freezing tolerance of the wood frog (Rana sylvatica), a northern woodland species that hibernates terrestrially in sites where dehydration and freezing may occur. Wood frogs inhabiting an outdoor enclosure accumulated urea to 65 mmol l-1 in autumn and early winter, when soil moisture was scarce, but subsequently urea levels fell to approximately 2 mmol l-1 as the availability of environmental water increased. Laboratory experiments showed that hibernating R. sylvatica can accumulate at least 90 mmol l-1 urea under relatively dry, warm conditions. During experimental freezing, frogs synthesized glucose but did not accumulate additional urea. Nevertheless, the concentrations of urea and glucose in some tissues were similar. We tested urea's efficacy as a cryoprotectant by measuring lysis and lactate dehydrogenase (LDH) leakage in samples of R. sylvatica erythrocytes frozen/thawed in the presence of physiological levels of urea or other osmolytes. In conferring protection against freeze/thaw damage, urea was comparable to glycerol and as good as or better than glucose, cryoprotectants found in freeze tolerant frogs and other animals. Urea treatment also improved the viability of intact tissues frozen in vitro, as demonstrated by post-thaw measures of metabolic activity and LDH leakage. Collectively, our findings suggest that urea functions both as an osmoprotectant and a cryoprotectant in terrestrially hibernating amphibians. PMID- 16244166 TI - Ionoregulatory changes in different populations of maturing sockeye salmon Oncorhynchus nerka during ocean and river migration. AB - We present the first data on changes in ionoregulatory physiology of maturing, migratory adult sockeye salmon Oncorhynchus nerka. Fraser River sockeye were intercepted in the ocean as far away as the Queen Charlotte Islands (approximately 850 km from the Fraser River) and during freshwater migration to the spawning grounds; for some populations this was a distance of over 700 km. Sockeye migrating in seawater toward the mouth of the Fraser River and upriver to spawning grounds showed a decline in gill Na+,K+-ATPase activity. As a result, gill Na+,K+-ATPase activity of fish arriving at the spawning grounds was significantly lower than values obtained from fish captured before entry into freshwater. Plasma osmolality and chloride levels also showed significant decreases from seawater values during the freshwater migration to spawning areas. Movement from seawater to freshwater increased mRNA expression of a freshwater specific Na+,K+-ATPase isoform (alpha1a) while having no effect on the seawater specific isoform (alpha1b). In addition, gill Na+,K+-ATPase activity generally increased in active spawners compared with unspawned fish on the spawning grounds and this was associated with a marked increase in Na+,K+-ATPase alpha1b mRNA. Increases in gill Na+,K+-ATPase activities observed in spawners suggests that the fish may be attempting to compensate for the osmotic perturbation associated with the decline in plasma chloride concentration and osmolality. PMID- 16244168 TI - Effects of intake rate on energy expenditure, somatic repair and reproduction of zebra finches. AB - Understanding the effect of food availability on food requirements is critical when linking food availability e.g. to reproduction or habitat selection. Decreasing intake rate (intake per unit foraging effort) can be expected to increase daily energy expenditure (DEE), due to increased foraging costs. However, all the studies we could find that have tested this hypothesis (with one exception) found DEE to be constant or decreasing when intake rate was experimentally decreased. This may be due to the design of the reward schedule, which can be fixed (e.g. 20 units effort required for each reward) or variable (e.g. each unit effort rewarded with probability 1/20). Most studies used fixed reward rates, but foraging motivation is generally higher for variable reward rates, and the only study in which animals increased DEE when intake rate decreased used variable reward rates. To assess the generality of this result, we exposed zebra finches Taeniopygia guttata to different intake rates using variable reward rates. We decreased intake rate by mixing 25 g of seeds with 0, 25 or 75 g of chaff. With increasing chaff/seed ratio the time spent foraging increased from 6% to 27%, but this was insufficient to compensate for the lower intake rate, because DEE decreased by 6.6%. Body mass was independent of chaff/seed ratio. Effects of intake rate on foraging time and DEE were stronger at lower temperatures, when DEE was higher. The decrease in DEE in adverse conditions raises the question of what prevents such behaviour in benign circumstances. We hypothesize that energy is saved at the expense of ;condition', and we tested this hypothesis in two ways. Firstly, we tested the effect of intake rate on the replacement of two plucked tail feathers (a form of somatic repair). Replacement feathers were shorter when intake rate was low, indicating an effect of intake rate on somatic repair ability. Secondly, we tested for carry over effects of intake rate on reproduction, by giving pairs the opportunity to reproduce with access ad libitum to food after feeding on one of the three chaff/seed ratios for 6 weeks. The interval until laying the first egg increased with decreasing intake rate in the preceding 6 weeks. The effects of intake rate on somatic maintenance and reproduction may explain why birds sustained higher metabolic rates than apparently necessary, but the physiological mechanisms determining the optimal metabolic rate remain to be discovered. PMID- 16244169 TI - Metabolic adjustments to increasing foraging costs of starlings in a closed economy. AB - Knowledge of the physiological consequences of variation in food availability may be essential for understanding behavioural and life history responses to such variation. To study the physiological consequences of food availability animals are generally subjected to caloric restriction or starvation, thereby reducing the upper limit to the energy budget. The relevance of this approach to free living animals is questionable, however, because under natural conditions low food availability often results in higher foraging costs, and everything else remaining equal this results in a higher energy budget. We manipulated food availability by varying the foraging costs and studied effects on daily energy expenditure (DEE) and energy allocation of captive starlings Sturnus vulgaris. Birds in a closed economy earned their food by flying between two perches 5 m apart. The probability of a reward was set at three different levels, thereby creating a 'poor', 'intermediate' and 'rich' environment. Compared with the rich environment, birds flew 4 times more (2.3 h per day) in the poor environment, and increased DEE by 43% to 220 kJ day-1 (3.7xBMR), within the range of free-living parents rearing young. To our knowledge this is the first study to show an increase in DEE with decreasing food availability. Body mass, basal metabolic rate (BMR) and pectoral muscle size were reduced in the poor environment. Nocturnal energy expenditure was further reduced by reaching BMR earlier in the night. Calculations show that the energy demands in the poor environment could not be met with the flight costs of 20.5 W that we measured previously in a rich environment. Flight costs derived indirectly from the energy budget were lower, at 17.5 W, probably due to lower body mass. By reducing body mass by 20%, and economising during sleep, the birds achieved savings of 37% in their DEE. Without these savings, a DEE substantially higher than measured in free-living parents rearing young would be required to remain in energy balance. Surprisingly little data exist to verify whether free-living animals use the same tactics to survive periods with low food availability. PMID- 16244170 TI - Mitochondrial depolarization following hydrogen sulfide exposure in erythrocytes from a sulfide-tolerant marine invertebrate. AB - Sulfide-tolerant marine invertebrates employ a variety of mechanisms to detoxify sulfide once it has entered their bodies, but their integumentary, respiratory epithelium and circulatory cells may still be exposed to toxic sulfide concentrations. To investigate whether sulfide exposure is toxic to mitochondria of a sulfide-tolerant invertebrate, we used the fluorescent dyes JC-1 and TMRM to determine the effect of sulfide exposure on mitochondrial depolarization in erythrocytes from the annelid Glycera dibranchiata. In erythrocytes exposed to 0.11-1.9 mmol l-1 sulfide for 1 h, the dyes showed fluorescence changes consistent with sulfide-induced mitochondrial depolarization. At the highest sulfide concentration, the extent of depolarization was equivalent to that caused by the mitochondrial uncoupler carbonyl cyanide m-chlorophenylhydrazone (CCCP). Even when induced by as little as 0.3 mmol l-1 sulfide, the depolarization was not reversible over a subsequent 5 h recovery period. The mechanism of toxicity was likely not via inhibition of cytochrome c oxidase (COX), since other COX inhibitors and other mitochondrial electron transport chain inhibitors did not produce similar effects. Furthermore, pharmacological inhibition of the mitochondrial permeability transition pore failed to prevent sulfide-induced depolarization. Finally, increased oxidation of the free radical indicators H2DCFDA and MitoSOX in erythrocytes exposed to sulfide suggests that sulfide oxidation increased oxidative stress and superoxide production, respectively. Together, these results indicate that sulfide exposure causes mitochondrial depolarization in cells of a sulfide-tolerant annelid, and that this effect, which differs from the actions of other COX inhibitors, may be via increased free radical damage. PMID- 16244172 TI - Determination of pH by microfluorometry: intracellular and interstitial pH regulation in developing early-stage fish embryos (Danio rerio). AB - Microfluorometric techniques were applied in vivo for continuous monitoring of specific acid-base parameters in zebrafish (Danio rerio) embryos during early stages of ontogeny. Dextran-coupled pH-sensitive single-excitation/dual-emission dye SNARF-1 was pressure-injected into individual cells or the interstitial space of 16- to 256-cell embryos, and pH was continuously recorded during subsequent development for time periods of up to 8 h. A novel calibration technique was developed, essentially characterized by in vitro inorganic buffer calibration of the optical system and mathematical post-processing according to the effects of in vivo dye modifiers through a correlation established by direct comparison of optical techniques with pH microelectrodes. This approach results in high accuracy of microfluorometry, comparable with that of pH electrodes, and a recovery only limited by the physical stability of the utilized optical system. Intracellular pH (pHi) in Danio rerio embryos between 1k-cells stage and the end of epiboly was found to be well regulated to a mean value of 7.55+/-0.13 (+/ s.d.), a range distinctly more alkaline than typical values for adult fish but in accordance with embryonic pHi of a few non-fish species shortly after fertilization. Also, interstitial pH (pHint) was significantly higher (8.08+/ 0.25) than values for extracellular pH in adult fish. Distributions of HCO3- across membranes and between interstitium and ambient fluid compared with respective potentials strongly suggest that pH in these early stages of ontogeny is already adjusted by active transfer processes. Non-respiratory changes in ambient pH between 7.7 and 8.5 did not significantly affect pHi, a result potentially attributable to low membrane leakage rate or to the potency of active transfer mechanisms. In order to assess the pH regulatory systems more quantitatively, embryos were exposed to ambient changes of carbon dioxide partial pressure (PCO2). The direct impact of PCO2 changes on cell pH was alleviated by cell non-bicarbonate buffering and subsequent rapid, almost complete, compensation by changes in cell [HCO3-] as an expression of transmembrane transfer of acid-base relevant ions. On the basis of these results, we conclude that the regulatory potency of embryonic cells is well developed, is active to resist extensive homoiostatic stress and is efficient to maintain critical metabolism in adverse conditions, even at early stages of ontogeny. PMID- 16244171 TI - Interactions of visual odometry and landmark guidance during food search in honeybees. AB - How do honeybees use visual odometry and goal-defining landmarks to guide food search? In one experiment, bees were trained to forage in an optic-flow-rich tunnel with a landmark positioned directly above the feeder. Subsequent food search tests indicated that bees searched much more accurately when both odometric and landmark cues were available than when only odometry was available. When the two cue sources were set in conflict, by shifting the position of the landmark in the tunnel during test, bees overwhelmingly used landmark cues rather than odometry. In another experiment, odometric cues were removed by training and testing in axially striped tunnels. The data show that bees did not weight landmarks as highly as when odometric cues were available, tending to search in the vicinity of the landmark for shorter periods. A third experiment, in which bees were trained with odometry but without a landmark, showed that a novel landmark placed anywhere in the tunnel during testing prevented bees from searching beyond the landmark location. Two further experiments, involving training bees to relatively longer distances with a goal-defining landmark, produced similar results to the initial experiment. One caveat was that, with the removal of the familiar landmark, bees tended to overshoot the training location, relative to the case where bees were trained without a landmark. Taken together, the results suggest that bees assign appropriate significance to odometric and landmark cues in a more flexible and dynamic way than previously envisaged. PMID- 16244173 TI - Retinoid cycles in the cone-dominated chicken retina. AB - In past decades, the role of retinoids in support of rod photopigment regeneration has been extensively characterized. In the rhodopsin cycle, retinal chromophore from bleached rod pigments is reduced to retinol and transferred to the retinal pigment epithelium (RPE) to store as all-trans retinyl ester. This ester pool is subsequently utilized for visual pigment regeneration. However, there is a lack of information on the putative cone visual cycle. In the present study, we provide experimental evidence in support of a novel retinoid cycle for cone photopigment regeneration. In the cone-rich chicken, light exposure resulted in the accumulation of 11-cis retinyl esters to the retina and all-trans retinyl esters to the RPE. Both the rate of increase and the amount of 11-cis retinyl esters in the retina far exceeded those of the all-trans retinyl esters in the RPE. In response to dark adaptation, this 11-cis retinyl ester pool in the retina depletes at a rate several times faster than the all-trans retinyl ester pool in the RPE. In vitro, isolated, dark-adapted retinas devoid of RPE show both an accumulation of 11-cis retinyl ester and a concomitant reduction of 11-cis retinal chromophore in response to light exposure. Finally, we provide experimental results to elucidate a cone visual cycle in chicken by relating the change in retinoids (retinal and retinyl ester) with time during light and dark adaptation. Our results support a new paradigm for cone photopigment regeneration in which the 11-cis retinyl ester pool in the retina serves as the primary source of visual chromophore for cone pigment regeneration. PMID- 16244174 TI - The contribution of tympanic transmission to fine temporal signal evaluation in an ultrasonic moth. AB - In lesser waxmoths Achroia grisella, pair formation and female mate choice involve very fine discrimination of male ultrasonic signals. Female A. grisella prefer male signals with longer pulses and longer ;asynchrony intervals', and evaluate differences in these characteristics in the range of 80-260 mus. The first step in the evaluation of these characteristics is the tympanic transmission of stimuli. We used laser vibrometry to describe the mode of vibration, frequency tuning and stimulus transmission of the tympana of A. grisella. The tympanic response consisted of a rotational mode of vibration, in which the anterior and posterior sections moved out of phase; the posterior section of the tympanum vibrated with all points moving in phase and maximum displacement at the attachment point of the scoloparium that contains the receptor cells. The tympana of A. grisella were tuned to high ultrasonic frequencies and had an estimated time constant (i.e. the limit to their temporal acuity) of about 20-50 mus. Pulse length and all but the shortest asynchrony interval were thus well resolved by the tympanum. We discuss implications for the evaluation of pulse length and asynchrony interval. PMID- 16244175 TI - Calcium-activated potassium channel of the tobacco hornworm, Manduca sexta: molecular characterization and expression analysis. AB - Large-conductance calcium- and voltage-gated potassium channels (BK or Slowpoke) serve as dynamic integrators linking electrical signaling and intracellular activity. These channels can mediate many different Ca2+-dependent physiological processes including the regulation of neuronal and neuroendocrine cell excitability and muscle contraction. To gain insights into the function of BK channels in vivo, we isolated a full-length cDNA encoding the alpha subunit of a Slowpoke channel from the tobacco hornworm, Manduca sexta (msslo). Amino acid sequence comparison of the deduced Manduca protein revealed at least 80% identity to the insect Slo channels. The five C-terminal alternative splice regions are conserved, but the cloned cDNA fragments contained some unique combinations of exons E, G and I. Our spatial profile revealed that transcript levels were highest in skeletal muscle when compared with the central nervous system (CNS) and visceral muscle. The temporal profile suggested that msslo expression is regulated developmentally in a tissue- and regional-specific pattern. The levels of msslo transcripts remain relatively constant throughout metamorphosis in the CNS, transiently decline in the heart and are barely detectable in the gut except in adults. A dramatic upregulation of msslo transcript levels occurs in thoracic but not abdominal dorsal longitudinal body wall muscles (DLM), suggesting that the msSlo current plays an important role in the excitation or contractile properties of the phasic flight muscle. Our developmental profile of msslo expression suggests that msSlo currents may contribute to the changes in neural circuits and muscle properties that produce stage-specific functions and behaviors. PMID- 16244176 TI - Hearing measurements from a stranded infant Risso's dolphin, Grampus griseus. AB - An infant Risso's dolphin (Grampus griseus) was rescued from the beach in Southern Portugal, and an audiogram was measured using auditory evoked potentials (AEP) and envelope following response (EFR) techniques for frequencies from 4 to 150 kHz. The stimuli used were custom sinusoidally amplitude-modulated (SAM) tone bursts, and the AEP responses were collected, averaged and analyzed to quantify the animal's physiological response and, thereby, hearing thresholds. The infant animal showed a wide range of best sensitivity, with the lowest threshold of 49.5 dB re. 1 microPa at 90 kHz. The audiogram showed a typical mammalian union or logical sum-shape with a gradual, low-frequency slope of 16.4 dB octave-1 and a sharp high-frequency increase of 95 dB octave-1. When compared with an audiogram of an older Risso's dolphin obtained using behavioral methods, the threshold values at upper frequencies were much lower for this infant animal, and this infant heard higher frequencies. These results redefine the hearing capabilities of Risso's dolphins by demonstrating very high-frequency sensitivity. PMID- 16244177 TI - Activation and inhibition of kidney CLC-K chloride channels by fenamates. AB - CLC-K Cl(-) channels are selectively expressed in kidney and ear, where they are pivotal for salt homeostasis, and loss-of-function mutations of CLC-Kb produce Bartter's syndrome type III. The only ligand known for CLC-K channels is a derivative of the 2-p-chlorophenoxypropionic acid (CPP), 3-phenyl-CPP, which blocks CLC-Ka, but not CLC-Kb. Here we show that in addition to this blocking site, CLC-K channels bear an activating binding site that controls channel opening. Using the voltage-clamp technique on channels expressed in Xenopus laevis oocytes, we found that niflumic acid (NFA) increases CLC-Ka and CLC-Kb currents in the 10 to 1000 microM range. Flufenamic acid (FFA) derivatives or high doses of NFA produced instead an inhibitory effect on CLC-Ka, but not on CLC Kb, and on blocker-insensitive CLC-Ka mutants, indicating that the activating binding site is distinct from the blocker site. Evaluation of the sensitivity of CLC-Ka to derivatives of NFA and FFA together with a modeling study of these ligands allow us to conclude that one major characteristic of activating compounds is the coplanarity of the two rings of the molecules, whereas block requires a noncoplanar configuration. These molecules provide a starting point for identification of diuretics or drugs useful in the treatment of Bartter's syndrome. PMID- 16244178 TI - Microtubule-associated protein 1B-light chain 1 enhances activation of Rap1 by exchange protein activated by cyclic AMP but not intracellular targeting. AB - We have previously demonstrated that EPAC1 interacts with light chain (LC) 2 of microtubule-associated protein (MAP) 1A. In the present study, we investigated whether the structurally related LC1 of MAP1B also interacts with EPAC1. We demonstrate that LC1 copurifies with EPAC1 from extracts of PC-12 cells, using cyclic AMP-agarose. Using recombinant LC1 and LC2 in pull-down and solid phase binding assays, we demonstrate direct interaction with a glutathione S transferase-fusion of the cyclic AMP-binding (CAMP) domain of EPAC1. We also tested whether LC1 directed intracellular targeting of EPAC1 through its interaction with the CAMP domain. EPAC1 was found be in the soluble and particulate, nuclear/perinuclear fractions of cells. We found that the catalytic (CAT) domain of EPAC1, and not the CAMP domain, was responsible for recruitment to the nuclear/perinuclear fraction of cells. The targeting sequence responsible was located between amino acids 764 and 838 of EPAC1. Overexpresssion of an isolated CAT domain in COS1 cells was found to displace endogenous EPAC1 from the nuclear/perinuclear fraction, thereby inhibiting EPAC-activated Rap1 in this compartment. In contrast, LC1 was not able to compete for the binding of EPAC1 to this fraction. LC1, however, was able to enhance interaction of EPAC1 with cyclic AMP and heightened the ability of EPAC to activate Rap1. Antibody disruption of EPAC1/LC1 interaction in PC-12 cells ablated the ability of cyclic AMP to activate Rap1. LC1 is therefore not involved in intracellular targeting of EPAC1, but it is rather a molecular chaperone of EPAC activity toward Rap1. PMID- 16244179 TI - Constitutive formation of oligomeric complexes between family B G protein-coupled vasoactive intestinal polypeptide and secretin receptors. AB - Formation of oligomeric complexes of family A G protein-coupled receptors has been shown to influence their function and regulation. However, little is known about the existence of such complexes for family B receptors in this superfamily. We previously used bioluminescence resonance energy transfer (BRET) to demonstrate that the prototypic family B secretin receptor forms ligand independent oligomeric complexes. Here, we show that subtypes of human vasoactive intestinal polypeptide receptors (VPAC1 and VPAC2) that represent the closest structurally related receptors to the secretin receptor also form constitutive oligomers with themselves and with the secretin receptor. We prepared tagged constructs expressing Renilla reniformis luciferase, yellow fluorescent protein, or cyan fluorescent protein at the carboxyl terminus of VPAC1, VPAC2, and secretin receptors, and performed BRET and morphologic fluorescence resonance energy transfer (FRET) studies with all combinations. The specificity of the BRET and FRET signals was confirmed by control studies. These constructs bound their natural ligands specifically and saturably, with these agonists able to elicit full cAMP responses. BRET studies showed that, like the secretin receptor, both VPAC receptors exhibited constitutive homo-oligomerization in COS cells. Unlike secretin receptor oligomers that were unaffected by ligand binding, the VPAC receptor homo-oligomers were modulated by vasoactive intestinal polypeptide. In addition, each of these three receptors formed hetero-oligomers with each other. The VPAC1-VPAC2 hetero-oligomers were modulated by vasoactive intestinal polypeptide binding, whereas the secretin-VPAC1 and secretin-VPAC2 receptor hetero-oligomers were unaffected by ligand treatment. Morphologic FRET studies demonstrated that each of the homo-oligomers and the VPAC1-VPAC2 receptor hetero oligomers reached the cell surface, where receptor interactions were clear. However, coexpression of secretin receptors with either type of VPAC receptor resulted in intracellular trapping of the hetero-oligomeric complexes within the biosynthetic pathway. These studies provide new insight into the ability of family B G protein-coupled receptors to associate with each other in cells. PMID- 16244180 TI - Sit to stand transfer: performance in rising power, transfer time and sway by age and sex in senior athletes. AB - OBJECTIVE: To observe the differences in performance variables of the sit to stand transfer (as measured on the NeuroCom Balance Master) in a population of senior athletes. METHOD: A convenience sample of 173 subjects aged 50 years and older. Data were obtained from voluntary participation in a health fair offered at the annual Huntsman World Senior Games in St George, Utah, USA. All sit to stand tests were performed on the NeuroCom Balance Master. The measured parameters were weight transfer time (WTT), rising power (force exerted to rise), and centre of gravity sway (COG sway) during the rising phase. RESULTS: A significant difference was found between stratified age groups (50-64 and 65+ years) on rising power. There was also a sex difference in rising power. No significant differences were found in weight transfer time or COG sway. CONCLUSION: While rising power decreases with increasing age in senior athletes, WTT and COG sway remain similar regardless of age or sex. The maintenance of these other two variables (WTT and COG sway) may be attributable to physical activity and/or participation in sport. PMID- 16244181 TI - Thrower's fracture of the humerus with radial nerve palsy: an unfamiliar softball injury. AB - A fracture of the normal humerus in a healthy young adult most commonly results from significant direct trauma. Throwing sports have become increasingly popular outside of North America and bring with them a novel injury mechanism for clinicians. A 21 year old woman sustained a "thrower's fracture" of the distal humerus and radial nerve palsy while throwing a softball. She was treated by internal fixation. Her fracture united, and radial nerve neurapraxia resolved after 8 weeks. Clinicians should be aware of this entity so that prodromal symptoms can be recognised early and thrower's fractures are not investigated unnecessarily. PMID- 16244182 TI - The time lords--measurement and performance in sprinting. PMID- 16244183 TI - Definitions for the purist. PMID- 16244184 TI - Research on mechano growth factor: its potential for optimising physical training as well as misuse in doping. PMID- 16244185 TI - Matrix metalloproteases: a role in overuse tendinopathies. PMID- 16244186 TI - Can exercise improve self esteem in children and young people? A systematic review of randomised controlled trials. AB - A systematic review to determine if exercise alone or as part of a comprehensive intervention can improve self esteem in children and young people is described. Twenty three randomised controlled trials were analysed. A synthesis of several small, low quality trials indicates that exercise may have short term beneficial effects on self esteem in children and adolescents. However, high quality research on defined populations with adequate follow up is needed. PMID- 16244187 TI - Muscle activity during the golf swing. AB - In the right hands, the golf swing is a motion that inspires looks of awe from the public. It is a complex movement of the whole body to generate power to a golf ball to propel the ball great distances with accuracy. This movement relies on the coordinated sequence of muscle activation to produce a fluid and reproducible movement. This paper reviews the literature on golf swing related muscle activity. The phases of this activity are discussed with a view to assisting the practitioner in understanding the swing. Such understanding may help in the management of the injured golfer. PMID- 16244188 TI - Detecting altered postural control after cerebral concussion in athletes with normal postural stability. AB - OBJECTIVE: To determine if approximate entropy (ApEn), a regularity statistic from non-linear dynamics, could detect changes in postural control during quiet standing in athletes with normal postural stability after cerebral concussion. METHODS: The study was a retrospective, case series analysis of centre of pressure (COP) data collected during the Sensory Organization Test (SOT) from NCAA Division I (USA) athletes prior to and within 48 h after injury. Subjects were 21 male and six female athletes from a variety of sports who sustained a cerebral concussion between 1997 and 2003. After injury, athletes displayed normal postural stability equivalent to preseason levels. For comparison, COP data also were collected from 15 male and 15 female healthy non-athletes on two occasions. ApEn values were calculated for COP anterior-posterior (AP) and medial lateral (ML) time series. RESULTS: Compared to healthy subjects, COP oscillations among athletes generally became more regular (lower ApEn value) after injury despite the absence of postural instability. For AP time series, declines in ApEn values were much larger in SOT conditions 1 and 2 (approximately three times as large as the standard error of the mean) than for all other conditions. For ML time series, ApEn values declined after injury in all sensory conditions (F(1,55) = 6.36, p = 0.02). CONCLUSIONS: Athletes who demonstrated normal postural stability after concussion nonetheless displayed subtle changes in postural control. Changes in ApEn may have represented a clinically abnormal finding. ApEn analysis of COP oscillations may be a valuable supplement to existing concussion assessment protocols for athletes. PMID- 16244189 TI - Rugby World Cup 2003 injury surveillance project. AB - OBJECTIVES: To study match injury patterns and incidence during the Rugby World Cup 2003 (RWC 2003); to compare these patterns and rates with comparative rugby injury data; and to assess differences between teams playing at different levels (eight finalists v 12 non-finalists). METHODS: Data were collected prospectively during the tournament. All injuries were recorded by the 20 participating team physicians. These were submitted to the tournament medical officer. An injury was defined as an event which forced a player either to leave the field or to miss a subsequent game or both. RESULTS: 189 injuries were recorded over 48 matches. This corresponds to 97.9 injuries per 1000 player-hours. Pool matches yielded a higher injury rate than non-pool matches. The 12 non-finalist teams sustained significantly higher injury rates than the eight finalist teams. The player positions open side flanker, inside centre, and number 8 were the most frequently injured positions. There was a low concussion rate, which may reflect under reporting. The non-finalist teams had a higher rate of recurrent injury. CONCLUSIONS: The injury rate was higher than comparative data. Mismatches in the areas of skill, fitness, and the availability of resources for medical care of players may explain these differences. PMID- 16244191 TI - Anthropometric and physiological profiles of sepak takraw players. AB - OBJECTIVES: Anthropometric and physiological profiles of national sepak takraw players were determined. METHODS: Thirty nine players, specialising in the three playing positions (tekong/server, feeder, and killer/spiker) were divided into three age categories of under 15 (U15), under 18 (U18), and under 23 (U23) years of age. Height, weight, percent body fat (%bf), maximum oxygen consumption (Vo(2max)), range of motion (ROM), back and leg strength, and heart rate, for the estimation of oxygen consumption during matches, were recorded. Statistical analysis was performed using one way ANOVA for independent measurements and data are presented as mean+/-standard deviation. RESULTS: The U23 players were significantly taller and heavier with significantly better ROM of the neck, trunk, and ankle joints and back and leg strength than the U15 players. No significant difference was found in %bf between the three age categories. Mean maximum heart rate during exercise was significantly higher in the U15 group when compared to the U18 and U23 groups (p<0.05). Mean Vo2max was similar between the three groups. Estimated oxygen consumption during matches was 69.1%, 68.5%, and 56.4% of Vo2max in the killer, tekong, and the feeder groups, respectively. CONCLUSIONS: The mean height, body weight, and cardiopulmonary capacities of the players were within the Malaysian population norms, but were somewhat lower than those of players of other court games from other countries. %bf was also lower in these players. This study provides the much needed anthropometric and physiological data of sepak takraw players for further development of this sport. PMID- 16244190 TI - Changes in joint position sense after surgically treated chronic lateral ankle instability. AB - BACKGROUND: A search of the literature shows that the effect of surgery on ankle proprioception has been hardly investigated. OBJECTIVE: To examine the effect of anatomical reconstruction of the anterolateral capsuloligamentous complex on ankle joint position sense. METHODS: A prospective study using the "slope box" test. Ten consecutive patients were included in the study, and 10 healthy athletes represented the control group. RESULTS: Similar test-retest reliability rates (overall reliability 0.92; p = 0.0013) were obtained to those of the original designers of the method. There were no significant differences with respect to side dominance (p = 0.9216). Investigation of the characteristics of mean absolute estimate errors showed that the controls tested became error prone in the range of slope altitudes 7.5-25 degrees in every direction, compared with the range 0-5 degrees (range of p values 0.00003-0.00072). The results of the intervention group showed that, for the two main directions of interest (anterior and lateral), preoperative differences in mean absolute estimate errors between injured (anterior 3.91 (2.81) degrees ; lateral 4.06 (2.85) degrees ) and healthy (anterior 2.94 (2.21) degrees , lateral 3.19 (2.64) degrees ) sides (anterior, p = 0.0124; lateral, p = 0.0250) had disappeared (postoperative differences: anterior, p = 0.6906; lateral, p = 0.4491). The afflicted ankle had improved significantly after surgery in both important directions (anterior, p<0.0001; lateral, p = 0.0023). CONCLUSIONS: The study shows that differences in joint position sense between healthy and injured ankles disappeared as the result of surgery. Preoperative data show that proprioceptive malfunction is a cause of functional instability. If treatment is by means of surgery, the retensioning of the original anterolateral structures is inevitable, even if other grafting or surgical techniques are used. PMID- 16244193 TI - Is postural control affected by expertise in alpine skiing? AB - OBJECTIVES: This study examined the postural performance of two groups of male skiers competing at different levels and the consequences on postural control of the suppression of visual afferences by eye closure. METHODS: Seven national level (NAT) skiers and 7 regional level (REG) skiers were asked to stand as still as possible on a force platform with eyes opened and closed and while wearing or not wearing their ski boots in a stable posture and in two unstable postures (in the sagittal or frontal plane). Postural performance was assessed with centre of foot pressure measurements. RESULTS: REG and NAT skiers were similarly influenced by the absence of visual information and presented similar postural performance when tests were performed with ski boots. However, without ski boots, REG skiers displayed better postural performance than NAT skiers. CONCLUSIONS: The inferior postural performance of NAT skiers without ski boots could be a long term effect of repetitive wearing of ski boots, which impairs postural performance by restricting the range of motion of the ankle-foot complex. Since individuals with decreased postural performance are believed to be more susceptible to ankle injury than those with finer postural control, NAT skiers should benefit from specific training aimed at improving postural ability and preventing ankle injury. PMID- 16244192 TI - Release of alpha-actin into serum after skeletal muscle damage. AB - OBJECTIVE: The skeletal muscle protein alpha-actin was investigated in the serum of subjects with severe skeletal muscle damage to assess its utility as a reliable and predictive marker of muscle damage. METHODS: Serum samples were obtained from 33 healthy controls and 33 patients with severe skeletal muscle damage, defined by a total creatine kinase value of >500 IU/l (Rosalki method). Troponin I, troponin T, and myoglobin concentrations were determined by immunoassay and alpha-actin concentrations by Western blot and densitometry. RESULTS: The mean serum concentration of alpha-actin in controls and patients with skeletal muscle damage was 600.9 and 1968.51 ng/ml, respectively, a statistically significant difference. Sera of patients with muscle damage showed higher levels of alpha-actin than of troponin or myoglobin. No significant difference in troponin I levels was observed between the groups. CONCLUSIONS: According to these results, alpha-actin was the most significant skeletal muscle damage marker analysed and may be an ideal candidate for the identification of all types of myofibre injury, including sports injuries. Our findings support the use of alpha-actin as a marker alongside other currently used biological proteins. PMID- 16244194 TI - Partial rupture of the proximal Achilles tendon: a differential diagnostic problem in ultrasound imaging. AB - OBJECTIVES: The aim of this study was to determine whether ultrasound can correctly visualise partial ruptures of the proximal Achilles tendon. METHOD: This was a prospective study in which all chronic Achilles tendon injury patients seen at three centres in Germany from 1998 to 2003 were screened. All patients with clinical and/or sonographic signs of abnormalities in the region of the proximal third of the Achilles tendon and tendomuscular junction were included in the analysis. Each of these cases was evaluated by ultrasound following an assessment protocol. Patients with ambiguous ultrasound findings and/or clinical signs were additionally assessed by magnetic resonance imaging (MRI). RESULTS: Sonomorphologic changes suggestive of an abnormality in the proximal third of the Achilles tendon were detected in 13 out of 320 patients (4.2%) with recurring Achilles tendon complaints. Thirteen patients had clinical signs but no sonographic changes in the tendon. The sonographic diagnosis was correct in 19 cases. In six of the 26 cases studied, MRI was needed to establish the correct diagnosis of partial intratendinous rupture of the proximal Achilles tendon. Sensitivity was 0.5, specificity was 0.81, and the overall agreement of the ultrasound examination was 61.5%. All patients were asymptomatic at follow up at a mean of 14 months (range 12-17 months) after surgery. CONCLUSIONS: Ultrasound is a useful tool for evaluation of proximal Achilles tendon complaints. However, ultrasound is not sufficiently reliable for diagnosis of all pathologies, especially partial ruptures of the Achilles tendon. Thus, the definitive diagnosis must be established by MRI. PMID- 16244195 TI - Is bowling workload a risk factor for injury to Australian junior cricket fast bowlers? AB - OBJECTIVES: To examine whether bowling workload is a risk factor for overuse injury to Australian junior cricket fast bowlers and to evaluate the appropriateness of current bowling workload guidelines. METHODS: Forty four male fast bowlers (mean (standard deviation) age 14.7 (1.4) years) were monitored prospectively over the 2002-2003 season. Bowlers completed a daily diary to record bowling workloads and self reported injuries, which were validated by a physiotherapist. Bowling workload prior to the first injury (for those bowlers who were injured) was compared to workload across the whole season for uninjured bowlers. RESULTS: Eleven (25%) bowlers reported an overuse-type injury, with seven of these sustaining a back injury. Injured bowlers had been bowling significantly more frequently than uninjured bowlers (median number of days since the previous bowling day: 3.2 v 3.9 days, Mann-Whitney U = 105.0, p = 0.038). Compared with bowlers with an average of > or =3.5 rest days between bowling, bowlers with an average of <3.5 rest days were at a significantly increased risk of injury (risk ratio (RR) = 3.1, 95% confidence interval (CI) 1.1 to 8.9). There were also trends towards an increased risk of injury for those who bowled an average of > or =2.5 days per week (RR = 2.5, 95% CI 0.9 to 7.4) or > or =50 deliveries per day (RR = 2.0, 95% CI 0.7 to 5.4). CONCLUSIONS: This study has identified high bowling workload as a risk factor for overuse injury to junior fast bowlers. Continued research is required to provide scientific evidence for bowling workload guidelines that are age-specific for junior fast bowlers. PMID- 16244196 TI - Superior results with eccentric compared to concentric quadriceps training in patients with jumper's knee: a prospective randomised study. AB - BACKGROUND: A recent study reported promising clinical results using eccentric quadriceps training on a decline board to treat jumper's knee (patellar tendinosis). METHODS: In this prospective study, athletes (mean age 25 years) with jumper's knee were randomised to treatment with either painful eccentric or painful concentric quadriceps training on a decline board. Fifteen exercises were repeated three times, twice daily, 7 days/week, for 12 weeks. All patients ceased sporting activities for the first 6 weeks. Age, height, weight, and duration of symptoms were similar between groups. Visual analogue scales (VAS; patient estimation of pain during exercise) and Victorian Institute of Sport Assessment (VISA) scores, before and after treatment, and patient satisfaction, were used for evaluation. RESULTS: In the eccentric group, for 9/10 tendons patients were satisfied with treatment, VAS decreased from 73 to 23 (p<0.005), and VISA score increased from 41 to 83 (p<0.005). In the concentric group, for 9/9 tendons patients were not satisfied, and there were no significant differences in VAS (from 74 to 68, p<0.34) and VISA score (from 41 to 37, p<0.34). At follow up (mean 32.6 months), patients in the eccentric group were still satisfied and sports active, but all patients in the concentric group had been treated surgically or by sclerosing injections. CONCLUSIONS: In conclusion, eccentric, but not concentric, quadriceps training on a decline board, seems to reduce pain in jumper's knee. The study aimed to include 20 patients in each group, but was stopped at the half time control because of poor results achieved in the concentric group. PMID- 16244197 TI - Changes in performance, skinfold thicknesses, and fat patterning after three years of intense athletic conditioning in high level runners. AB - OBJECTIVES: To determine if the changes in specific skinfold sites induced by intense athletic conditioning over a three year period were associated with changes in running performance in high level athletes. METHODS: Thirty seven top class runners (eight male and five female sprint trained, 16 male and eight female endurance trained) volunteered to participate in the study. The athletes were divided into class A (n = 18) and class B (n = 17), with class A having the best performance. Biceps, triceps, subscapular, pectoral, iliac crest, abdominal, front thigh, and medial calf skinfold thickness and the best running performance were recorded at the beginning and after one, two, and three years of training. A one way analysis of variance and a linear regression analysis were conducted to determine changes and association between performance and skinfold thicknesses. Analyses were controlled for sex, sprint event or endurance event, and class. RESULTS: Training resulted in a significant increase in performance and decreases in sum of six skinfolds, abdominal, front thigh, and medial calf skinfolds, and the ratio of extremity to trunk skinfolds (E/T, Sigmatriceps, front thigh, medial calf/Sigmasubscapular, iliac crest, abdominal). There were no significant differences in body weight. Except for the abdominal skinfold, there was no significant difference in trunk skinfolds. Significant differences in these changes were observed by sex for E/T, which decreased and increased in male and female runners respectively, and by class. Class B runners significantly improved performance, with decreased skinfold thicknesses in the lower limb. There were no significant changes in performance or skinfold thicknesses in class A runners. Improvements in performance were consistently associated with a decrease in the lower limb skinfolds. CONCLUSIONS: On the basis of these findings, anthropometric assessment of top class athletes should include an evaluation of all skinfolds. The loss of body fat appears to be specific to the muscular groups used during training. The lower limb skinfolds may be particularly useful predictors of running performance. PMID- 16244198 TI - Early, active rehabilitation following mini-open repair of Achilles tendon rupture: a prospective study. AB - OBJECTIVE: To assess the use of a supervised active rehabilitation program following repair of acute Achilles tendon ruptures using a minimally invasive suture system. METHODS: We performed a prospective study on 46 patients undergoing surgical repair of acute Achilles tendon ruptures using the Achillon suture system. All patients began a supervised active rehabilitation program from 2 weeks postoperatively. Patients were placed in a range of motion brace fixed at 20 degrees equinus for 2 weeks to allow wound healing followed by active movement from neutral to full plantar flexion for 4 weeks. RESULTS: At a minimum follow up of 12 months there were no re-ruptures. All patients were able to return to their previous sporting activities by 6 months post operation. The average American Orthopaedic Foot and Ankle Society (AOFAS) score at 6 months was 98, with 42 patients having excellent and four patients good Leppilahti scores. The average time to return to work was 22 days. One patient had a superficial wound infection which settled with 5 days of oral antibiotics. Two patients had altered sensation in the distribution of the sural nerve which settled spontaneously within 3 months. CONCLUSION: The Achillon suture system appears to allow a safe early active rehabilitation program and achieves a high rate of success. Further evaluation is necessary with regard to potential damage to the sural nerve. PMID- 16244199 TI - Acute whole body vibration training increases vertical jump and flexibility performance in elite female field hockey players. AB - OBJECTIVE: To quantify the acute effect of whole body vibration (WBV) training on arm countermovement vertical jump (ACMVJ), grip strength, and flexibility performance. METHODS: Eighteen female elite field hockey players each completed three interventions of WBV, control, and cycling in a balanced random manner. WBV was performed on a Galileo machine (26 Hz) with six different exercises being performed. For the control, the same six exercises were performed at 0 Hz, whilst cycling was performed at 50 W. Each intervention was 5 min in duration with ACMVJ, grip strength, and flexibility measurements being conducted pre and post intervention. RESULTS: There was a positive interaction effect (intervention x pre-post) of enhanced ACMVJ (p<0.001) and flexibility (p<0.05) parameters following WBV; however no changes were observed after the control and cycling interventions. There was no interaction effect for grip strength following the three interventions. CONCLUSIONS: Acute WBV causes neural potentiation of the stretch reflex loop as shown by the improved ACMVJ and flexibility performance. Additionally, muscle groups less proportionally exposed to vibration do not exhibit physiological changes that potentiate muscular performance. PMID- 16244201 TI - Cobalt chloride administration in athletes: a new perspective in blood doping? AB - Blood doping is an illegal and unfair way of enhancing athletic performance by increasing the oxygen carrying capacity of the blood. Currently used methods usually involve stimulation of erythropoiesis. Gene therapy targeting the hypoxia inducible factor pathway may be an attractive alternative to traditional blood doping techniques. Hypoxia activates a large number of genes with essential roles in cell and tissue adaptation to low oxygen. Cobalt chloride is a well established chemical inducer of hypoxia-like responses such as erythropoiesis. Cobalt supplementation is not banned and therefore would not be detected by current anti-doping testing. Although there is as yet no direct or anecdotal evidence of cobalt chloride administration to athletes, its use should be warned against as being not only unfair but potentially dangerous. PMID- 16244202 TI - "Read with caution": a reply to Pickett et al. PMID- 16244200 TI - Measurement of functional recovery in individuals with acute anterior cruciate ligament rupture. AB - OBJECTIVES: To measure functional recovery following acute anterior cruciate ligament (ACL) rupture using a simple and reliable clinical movement analysis system. Clinic based methods that simultaneously quantify different aspects of movement over a range of activities and model functional recovery will help guide rehabilitation. METHODS: A longitudinal study was used to measure gait variables at initial physiotherapy attendance and then at monthly intervals using a digital camcorder and computer for quantitative analysis. Jogging and distance hopping were added during recovery. A sample of 63 ACL deficient subjects entered the study and 48 subjects were measured at least three times. To determine the pattern of recovery, repeated measurements were analysed using a least square fit of the data. RESULTS: Gait variables took between 95 and 130 days post injury to reach the control mean and stabilise shortly after this. Hopping distance for the injured leg took 62 days to recover to within normal limits and 5 months post injury to reach the control mean. Jogging was already within the control limits at 30 days post injury and demonstrated little change with recovery. CONCLUSIONS: Functional recovery of multiple variables has been modelled. In the early phase of post injury, gait velocity seems to be the most useful variable to measure improvement. Recovery of more challenging activities appears to take an average of 5 months. Therefore, patients may need to be monitored in physiotherapy until this time and advised not to return to sport until sufficient recovery is demonstrated on activities such as distance hopping. PMID- 16244203 TI - American Academy of Pediatrics guidelines for detecting neonatal hyperbilirubinaemia and preventing kernicterus. PMID- 16244204 TI - American Academy of Pediatrics guidelines for detecting neonatal hyperbilirubinaemia and preventing kernicterus. PMID- 16244205 TI - The car seat: a challenge too far for preterm infants? AB - Physiological monitoring studies have shown that some preterm infants who are otherwise ready for hospital discharge experience episodes of oxygen desaturation, apnoea, or bradycardia when seated in standard car safety seats. The American Academy of Pediatrics recommends that all preterm infants are assessed for cardiorespiratory stability in their car seat before discharge: the "car seat challenge". This screening test has been incorporated into discharge assessments in most neonatal units in North America and is being increasingly used in the United Kingdom and elsewhere. The evidence base for the use of the car seat challenge in discharge assessment, the possible implications for infants, their families, and health services of adopting the practice, and the issues that may be resolved with further research are discussed. PMID- 16244206 TI - Fit to fly: practical challenges in neonatal transfers by air. AB - Air transport of infants in incubators should be undertaken in a manner that is safe for both staff and infant and satisfies all directives by appropriate regulatory bodies. In Scotland during the last two years, certification of an infant incubator system for use in both rotary and fixed wing aircraft has been accomplished. This is a report of the issues addressed during this project, which will be common to all neonatal transport services intending to develop air transport capability. PMID- 16244208 TI - Watershed infarcts in the full term neonatal brain. PMID- 16244207 TI - Severe acute respiratory syndrome (SARS) in neonates and children. AB - Severe acute respiratory syndrome (SARS) runs a more benign course in children during the acute phase. Infants born to mothers with the disease did not acquire the infection through vertical transmission. The treatment strategy for children with SARS has not been standardised and is based on adult experience. Thus far, no deaths have been reported in the paediatric age group. Exercise impairment and residual radiological abnormalities were present six months after diagnosis. It is important to assess these patients on a regular basis to detect and provide appropriate management for any persistent or emerging long term sequelae in the physical, psychological, and social domains. This review describes the current understanding of SARS coronavirus infection in newborns and children. PMID- 16244209 TI - Does variation in interpretation of ultrasonograms account for the variation in incidence of germinal matrix/intraventricular haemorrhage between newborn intensive care units in New Zealand? AB - BACKGROUND: The incidence of germinal matrix/intraventricular haemorrhage (GM/IVH) reported to the Australian and New Zealand Neonatal Network (ANZNN) varies between neonatal intensive care units (NICUs). HYPOTHESIS: Differences in the capture, storage, and interpretation of the cerebral ultrasound scans may account for some of this variation. METHODS: A total of 255 infants with birth weight <1500 g and gestation <32 weeks born between 1997 and 2002 were randomly selected from the ANZNN database, 44 from each of the six NICUs in New Zealand. Twenty two infants from each NICU had cerebral ultrasound scans previously reported to ANZNN as normal; another 22 had scans reported as abnormal. The original scans were copied using digital photography and anonymised and independently read by a panel of three experts using a standardised method of reviewing and reporting. RESULTS: There was considerable variation between NICUs in methods of image capture and quality and completeness of the scans. However, there was little variation in the reporting of scans between the reviewers and the reports to ANZNN (weighted kappa 0.75-0.91). Grade 1 GM/IVH was generally over-reported and grade 4 under-reported to the ANZNN. CONCLUSION: For all NICUs, a high level of agreement was found between the reviewers' reports and the reports to the ANZNN. Thus the variation between NICUs in the incidence of GM/IVH reported to the ANZNN is unlikely to be due to differences in capture, storage, and interpretation of the cerebral ultrasound scans. Further investigation is warranted into the reasons for the variation in incidence of GM/IVH between NICUs. PMID- 16244210 TI - Temperature variation in newborn babies: importance of physical contact with the mother. AB - BACKGROUND: Hypothermia is a major cause of deterioration and death in the neonatal period. Temperature deviations are key signs of illness. OBJECTIVE: To determine normal patterns of temperature variation in newborn babies and the influence of external factors. METHODS: Abdominal and foot skin temperature were continuously recorded in 27 healthy full term babies during the first two days of life and related to the care situation-that is, whether the baby was with the mother or in its cot. The recordings were made using no wires to avoid interference with the care of the neonate. Ambient temperature was close to 23 degrees C during the study period. RESULTS: Mean rectal and abdominal and foot skin temperature were lower on day 1 than day 2. The foot skin temperature was directly related to the care situation, being significantly higher when the baby was with the mother. The abdominal skin temperature was much less influenced by external factors. When the neonates were with their mothers, the mean difference between rectal temperature and abdominal skin temperature was 0.2 degrees C compared with a mean difference between rectal temperature and foot skin temperature of 1.5 degrees C, indicating a positive heat balance. In the cot the corresponding temperature differences were 0.7 degrees C and 7.5 degrees C. A temperature difference between rectal and foot skin temperature of 7-8 degrees C indicates a heat loss close to the maximum for which a neonate can compensate (about 70 W/m2). CONCLUSION: This study emphasises the importance of close physical contact with the mothers for temperature regulation during the first few postnatal days. PMID- 16244211 TI - Potential risk factors for the development of acute renal failure in preterm newborn infants: a case-control study. AB - AIMS: To determine in a case-control study possible associations between the development of acute renal failure in preterm newborns and therapeutic interventions, particularly drug treatments. METHODS: The study population was 172 preterm infants of <38 weeks gestation; 71 had acute renal failure and 101 were controls closely matched for gestational age and birth weight. Maternal and neonatal information was collected for both groups through questionnaires and interviews. Routine data on renal variables were also collected. Univariate and multivariate logistic regression analyses were performed. RESULTS: Very low birthweight infants were at high risk of acute renal failure (79% of cases were <1500 g). However, the acute renal failure was transient. Mothers of infants with acute renal failure received more drugs during pregnancy and delivery (mainly antibiotics and non-steroidal anti-inflammatory drugs). Of the possible therapeutic interventions, intubation, catheterisation, and phototherapy were mainly applied to case subjects. A low Apgar score and patent ductus arteriosus were diagnosed in a greater percentage of neonates with acute renal failure. Moreover, in the first few days of life and before diagnosis of acute renal failure, case subjects received more drugs (antibiotics, non-steroidal anti inflammatory drugs, and diuretics) and for a longer time. In the multivariate logistic analysis, medullary hyperechogenicity (odds ratio (OR) 4.491; 95% confidence interval (CI) 1.879 to 10.731) and ceftazidime administration (OR 5.082; 95% CI 1.493 to 17.297) were associated with a greater risk of acute renal failure. CONCLUSIONS: The results suggest the need for careful monitoring of very low birthweight infants and attention to drug treatments, as it is difficult to differentiate between normality and renal failure in the first few days of life. PMID- 16244212 TI - Sildenafil in neonatal pulmonary hypertension due to impaired alveolarisation and plexiform pulmonary arteriopathy. AB - We report a case of severe pulmonary hypertension in a neonate associated with impaired alveolarisation and plexiform pulmonary arteriopathy. Treatment with oral sildenafil in addition to inhaled nitric oxide (NO) resulted in recovery from the pulmonary hypertensive crisis. Long term sildenafil therapy was associated with complete resolution of the pulmonary hypertension. PMID- 16244214 TI - Chitotriosidase in neonates with fungal and bacterial infections. AB - Increased plasma and/or urine chitotriosidase activity was found in neonates with fungal infection changing in parallel with their clinical condition. Increased levels were also found in neonates with bacterial infection. Chitotriosidase activity increase is not a response specific to fungi, but serial assays could monitor the course of neonatal fungal infection. PMID- 16244213 TI - Cytomegalovirus remains viable in naturally infected breast milk despite being frozen for 10 days. AB - Cytomegalovirus culture positive breast milk was obtained from four mothers of very premature babies. The milk was stored at 0-5 degrees C in a domestic refrigerator for 48 hours or frozen for different durations at -20 degrees C. Cytomegalovirus survived in breast milk despite being frozen for 10 days at -20 degrees C. PMID- 16244215 TI - Scald injury from the Guthrie test: should the heel be warmed? AB - A 6 day old boy sustained scalds to his right foot from application of a "warm nappy" to increase blood flow to facilitate heel pricking for a routine Guthrie test. It is proposed that active warming of the heel in isolation is not necessary in heel blood sampling. PMID- 16244216 TI - The giant umbilical cord. AB - The giant umbilical cord is a rare malformation of the umbilical cord that can easily be diagnosed on prenatal scans and is unmistakable postnatally. We report a case to highlight issues of this rare finding. Visual diagnosis is easy and surgical repair is usually required. PMID- 16244217 TI - Sir Robert Hutchison (1871-1960) of London and the causes and treatment of rickets. AB - A general physician with a special interest in nutrition and in children, Sir Robert Hutchison became a leader in medicine and paediatrics in the United Kingdom during the early decades of the 20th century. His wisdom and wit educated and amused successive generations of students and doctors. PMID- 16244218 TI - Are endocannabinoids the basis for neonatal analgesia through non-nutritive sucking? PMID- 16244219 TI - Is current management of neonatal jaundice evidence based? PMID- 16244220 TI - Two-photon fluorescence cross-correlation spectroscopy as a potential tool for high-throughput screening of DNA repair activity. AB - Several lines of evidence indicate that differences in DNA repair capacity are an important source of variability in cancer risk. However, traditional assays for measurement of DNA repair activity in human samples are laborious and time consuming. DNA glycosylases are the first step in base excision repair of a variety of modified DNA bases. Here, we describe the development of a new sensitive DNA glycosylase assay based on fluorescence cross-correlation spectroscopy (FCCS) with two-photon excitation. FCCS was applied to the measurement of uracil DNA glycosylase activity of human cell extracts and validated by comparison with standard gel electrophoresis assay. Our results indicate that FCCS can be adapted to efficient assays for DNA glycosylase activity in protein extracts from human cells. This method has a potential for the development of automated screening of large number of samples. PMID- 16244221 TI - Improving molecular cancer class discovery through sparse non-negative matrix factorization. AB - MOTIVATION: Identifying different cancer classes or subclasses with similar morphological appearances presents a challenging problem and has important implication in cancer diagnosis and treatment. Clustering based on gene expression data has been shown to be a powerful method in cancer class discovery. Non-negative matrix factorization is one such method and was shown to be advantageous over other clustering techniques, such as hierarchical clustering or self-organizing maps. In this paper, we investigate the benefit of explicitly enforcing sparseness in the factorization process. RESULTS: We report an improved unsupervised method for cancer classification by the use of gene-expression profile via sparse non-negative matrix factorization. We demonstrate the improvement by direct comparison with classic non-negative matrix factorization on the three well-studied datasets. In addition, we illustrate how to identify a small subset of co-expressed genes that may be directly involved in cancer. PMID- 16244223 TI - H5N1 avian influenza virus reaches Europe. PMID- 16244225 TI - Animal Welfare Bill laid before Parliament. PMID- 16244227 TI - Practice standards today... PMID- 16244231 TI - Discrimination of isolates of Mycobacterium bovis in Northern Ireland on the basis of variable numbers of tandem repeats (VNTRs). AB - The ability to reproducibly discriminate Mycobacterium bovis isolates and trace their transmission has the potential to clarify sources of infection and major routes of transmission for bovine tuberculosis (TB). A PCR-based genotyping assay has been developed to discriminate between strains of M bovis by examining multiple sites in its genome that consist of variable numbers of tandem repeats (VNTRS). The discriminatory power and reproducibility of this VNTR typing has been compared with that of the established PCR-based spoligotyping technique by using a panel of 461 isolates of M bovis prevalent in Northern Ireland. The VNTR assay discriminated 40 different profiles, the most prevalent of which constituted 21 per cent of the total, compared with 14 profiles discriminated by spoligotyping, the most prevalent of which constituted 65 per cent. No significant differences were observed between the prevalences of the VNTR profiles in the years from 1999 to 2003. A preliminary evaluation indicated that most genotypes predominated in particular areas of the country. This VTNR typing assay was found to be highly discriminating, with the performance characteristics to support its systematic application to the molecular epidemiology of bovine TB. PMID- 16244232 TI - Review of 15 cases of pemphigus foliaceus in horses and a survey of the literature. AB - The records of 15 horses with pemphigus foliaceus diagnosed on the basis of their history, clinical signs, histopathology and the exclusion of differential diagnoses were evaluated with respect to the age of onset, the clinical signs and the diagnostic tests used. There was no apparent breed predisposition. The horses' mean age was nine years, with a range from three months to 25.5 years, three were foals up to six months old and eight were nine years old or older. The most frequent lesions were scaling in 11, crusting in 10 and alopecia in 10, and they appeared most commonly on the face, neck and trunk, in 10 horses for each of these sites. The extremities were involved in nine of the horses, pruritus occurred in seven, and four of the horses had pustules. The clinical signs mostly corresponded with those described in previous reports, but signs of pain were not a prominent feature. Acantholytic cells were identified cytologically in four of six of the horses. PMID- 16244233 TI - Topical treatment of non-healing corneal epithelial ulcers in dogs with aminocaproic acid. AB - The potential efficacy of topical epsilon-aminocaproic acid, an antiplasmin agent, in the treatment of persistent corneal epithelial defects was evaluated in a study of the medical records of 44 dogs, in which 51 eyes had been diagnosed with a corneal epithelial defect lasting more than 10 days, with no apparent underlying cause. At an initial examination all the affected eyes had had non adherent epithelium removed. Thirty-four of the eyes in 28 dogs examined between January 2000 and March 2003 were also treated by the topical application of a solution of 35.7 mg/ml ophthalmic aminocaproic acid three times a day; the other 17 eyes in 16 dogs treated between October 1997 and March 1999 had received only topical treatment with gentamicin in addition to the debridement. Both groups were assessed clinically at weekly intervals for a maximum of three weeks. The two groups had approximately the same breed distribution, and there were no statistically significant differences between them in terms of their age, sex, affected side or duration of the corneal erosions. After three weeks, 32 of the 34 eyes treated with aminocaproic acid (94.1 per cent) had been cured, compared with seven of the 17 eyes treated with gentamicin (41.2 per cent) (P=0.0001). No adverse drug reactions were reported. PMID- 16244234 TI - Anatomical, histological and genetic investigations of a sexually anomalous goat. AB - An apparently female goat was masculine in appearance and had strong male-pattern libido and behaviour. The goat's DNA was subjected to PCR analysis and its reproductive system was examined anatomically and histologically. The external genitalia consisted of an apparent vulva in the normal position and a grossly enlarged clitoris. Two masses could be palpated lying subcutaneously in the inguinal region; when they were removed they had the macroscopic appearance of hypoplastic testes, and histologically they had atrophic tubules and prominent interstitial tissues. To exclude sex chimaerism, X and Y chromosome-specific targets were amplified by PCR, using bone marrow and muscle DNA templates. The goat had some Y chromosome-specific sequences such as SRY and BRY.I but lacked others, such as BOV97M. PMID- 16244235 TI - Coccidiosis in replacement gilts. PMID- 16244236 TI - Serological prevalence of Leptospira interrogans serovar Bratislava in horses in Mongolia. PMID- 16244237 TI - Diabetes mellitus in an adult alpaca (Lama pacos). PMID- 16244238 TI - Peste des petits ruminants in a sheep and goat flock in Kayseri province, Turkey. PMID- 16244239 TI - The Supply of Relevant Veterinary Medicinal Products Order 2005. PMID- 16244240 TI - Isolation and identification of avian influenza viruses from migratory birds in Iran. PMID- 16244241 TI - Testing for antibodies to equine arteritis virus. PMID- 16244242 TI - Canine influenza virus. PMID- 16244243 TI - Avian vibrionic hepatitis in laying hens. PMID- 16244244 TI - Toe-out conformation in thoroughbred horses in Sudan. PMID- 16244245 TI - Science to practice: can combination therapy for colorectal liver metastases improve results over chemoembolization or ablation alone? PMID- 16244246 TI - Responsible conduct of radiology research. Part IV. The boundary of research and practice. AB - The purpose of this series of articles is to explain the ethical and legal basis for responsible conduct of radiology research and the rules that an investigator must follow. Part four of this series explains special situations that commonly arise during human research in radiology. The boundaries of research and innovative care or quality improvement often are not clear. When human research activities are or become subject to regulation, they must be reviewed and approved by an institutional review board. Therefore, physicians performing innovative care or conducting quality improvement should have a low threshold for seeking institutional review board guidance. Physicians performing innovative care should also consider their ethical obligation to future patients to conduct research to prove the validity of such innovative care. PMID- 16244248 TI - Screening with multisection CT: unmasking the benefit. PMID- 16244247 TI - Guidelines for management of small pulmonary nodules detected on CT scans: a statement from the Fleischner Society. AB - Lung nodules are detected very commonly on computed tomographic (CT) scans of the chest, and the ability to detect very small nodules improves with each new generation of CT scanner. In reported studies, up to 51% of smokers aged 50 years or older have pulmonary nodules on CT scans. However, the existing guidelines for follow-up and management of noncalcified nodules detected on nonscreening CT scans were developed before widespread use of multi-detector row CT and still indicate that every indeterminate nodule should be followed with serial CT for a minimum of 2 years. This policy, which requires large numbers of studies to be performed at considerable expense and with substantial radiation exposure for the affected population, has not proved to be beneficial or cost-effective. During the past 5 years, new information regarding prevalence, biologic characteristics, and growth rates of small lung cancers has become available; thus, the authors believe that the time-honored requirement to follow every small indeterminate nodule with serial CT should be revised. In this statement, which has been approved by the Fleischner Society, the pertinent data are reviewed, the authors' conclusions are summarized, and new guidelines are proposed for follow-up and management of small pulmonary nodules detected on CT scans. PMID- 16244249 TI - Finding evidence-based answers to practical questions in radiology: which patients with inoperable hepatocellular carcinoma will survive longer after transarterial chemoembolization? AB - To some, evidence-based practice (EBP) means the identification of centers that produce evidence reports and technology assessments to support guideline development. To others, EBP is the best research evidence integrated with clinical expertise and patient values. Inherent in the first approach is the implication that only central academic organizations can produce valid, reliable analyses of existing literature, which will then be distributed to ordinary practitioners. The second approach implies that ordinary practitioners can learn to use a stepwise approach and a preprepared set of rules and tools to effectively find the best current literature, appraise it, and then apply local circumstances to these rules and tools in their hospital. Paul Glasziou, director of the Centre for Evidence-based Practice in Oxford, England, has coined the phrases top-down EBP and bottom-up EBP to describe these approaches. In this article, the authors describe how knowledge gaps in an ordinary radiology practice can be addressed by using stepwise bottom-up EBP techniques. The following clinical scenario is used: Your hospital's recently appointed chief hepatobiliary surgeon questions the use of transarterial chemoembolization for inoperable hepatocellular carcinoma because of his concerns after reading a recent review article suggesting that there is no clear survival benefit to using this procedure. What would you do? Here is how the authors would do it. PMID- 16244250 TI - The lactating breast: contrast-enhanced MR imaging of normal tissue and cancer. AB - PURPOSE: To retrospectively describe the magnetic resonance (MR) imaging characteristics of normal breast tissue and breast cancer in the setting of lactation. MATERIALS AND METHODS: The HIPAA-compliant study was exempt from institutional approval, and informed consent was not required. Unilateral MR imaging of 10 breasts was performed in seven lactating patients aged 27-42 years. For the three patients in whom both breasts were imaged, each breast was imaged on a separate day. Nonenhanced T1-weighted and fat-saturated T2-weighted images and contrast material-enhanced dynamic three-dimensional (3D) T1-weighted spiral gradient-echo images interleaved with T1-weighted high-spatial-resolution 3D gradient-echo images (2.0 x 1.0 x 0.4-mm voxels) were obtained. Three readers in consensus assessed the glandular density, T2-weighted signal intensity, milk duct appearance, and contrast enhancement in normal and tumor-containing breast regions. The pharmacokinetic contrast enhancement parameters of tumors were compared with those of normal tissue by using Student t and Mann-Whitney tests. RESULTS: MR findings of normal breast tissue in the seven women included increased glandular density in six women, high T2-weighted signal intensity in six, dilated central ducts in seven, and rapid initial glandular contrast enhancement in seven. MR findings of invasive ductal carcinoma in five women, compared with findings of the normal glandular tissue, included lower T2-weighted signal intensity in five women, more avid and rapid contrast enhancement in five, and early contrast enhancement washout in four. One minute after contrast agent injection, tumor signal intensity increased significantly more than normal lactating tissue signal intensity (153% vs 60% from baseline, P = .016). The median two-compartment model K(21) exchange rate in the tumors, 0.078 sec(-1), was significantly faster than the K(21) exchange rate in normal tissue, 0.011 sec(-1) (P = .03). CONCLUSION: Normal lactating glands have increased density, high T2-weighted signal intensity, and rapid moderate contrast enhancement. Breast cancers are visible during lactation owing to their lower signal intensity and more intense initial contrast enhancement with early washout compared with normal breast tissue. PMID- 16244251 TI - Influence of review design on percentages of missed interval breast cancers: retrospective study of interval cancers in a population-based screening program. AB - PURPOSE: To retrospectively investigate whether different review designs have an influence on the estimate of missed interval cancer in a population-based breast cancer screening program. MATERIALS AND METHODS: The Norwegian Breast Cancer Screening Program invites women aged 50-69 years to undergo biennial screening mammography. The current study was part of the evaluation and scientific aspects of the screening program and thus was covered by the general ethical approval of the screening program as a part of the Cancer Registry of Norway. All participants signed an informed consent that specified that data related to their screening visit could be used for evaluation and scientific purposes. Six radiologists (9-34 years of experience in mammography) reviewed previously obtained bilateral two-view screening and diagnostic mammograms of 231 interval cancers, 117 screening-detected cancers, and 373 normal cases. Four review designs were used: individual and paired blinded review and individual and consensus informed review. A five-point interpretation scale was used to reclassify the cancers into missed cancers, minimal signs, and true cancers. The number and proportion of subgroups were estimated with 95% confidence intervals. RESULTS: Of 231 interval cancers, 46 (19.9%) were reclassified as missed cancers with the mixed blinded individual review and 54 (23.4%) were classified as missed cancers with the mixed blinded paired review. Eighty-three cancers (35.9%) were classified as missed cancers with individual informed review, and 78 (33.8%) were classified as missed cancers with consensus informed review. Thirty-nine cancers (16.8%) were reclassified as missed when four or more radiologists assigned a score of 2 or more (probably benign or more suspicious); three cancers (1.3%) were reclassified as missed when a score of 4 or more (probably malignant or more suspicious) was assigned. CONCLUSION: The percentage of interval cancers classified as missed ranged from 1.3% to 35.9% according to review design. To encourage learning, a review protocol should include both blinded and informed designs. PMID- 16244252 TI - Computer-aided detection in the United Kingdom National Breast Screening Programme: prospective study. AB - PURPOSE: To evaluate prospectively the recall and cancer detection rates with and without computer-aided detection (CAD) in the United Kingdom National Health Service Breast Screening Programme. MATERIALS AND METHODS: The study had appropriate ethics committee approval. Informed consent was not required; however, patients were informed that their mammograms might be used in research efforts, and all patients agreed to participate. Mammograms obtained in 6111 women (mean age, 58.4 years) undergoing routine screening every 3 years were analyzed with a CAD system. Mammograms were independently double read. Twelve readers participated. Readers recorded an initial evaluation, viewed the CAD prompts, and recorded a final evaluation. Recall to assessment was decided after arbitration. Sensitivities were calculated for single reading, single reading with CAD, and double reading, as a proportion of the total number of cancers detected by using double reading with CAD. RESULTS: A total of 62 cancers were detected in 61 women. CAD prompted 51 (84%) of 61 radiographically detected cancers. Of 12 cancers missed on single reading, nine were correctly prompted; however, seven prompts were overruled by the reader. Sensitivity of single reading was 90.2% (95% confidence interval [CI]: 83.0%, 95.0%), single reading with CAD was 91.5% (95% CI: 85.0%, 96.0%), and double reading without CAD was 98.4% (95% CI: 91.0%, 100%). Cancer detection rate was 1%. Recall to assessment rate was 6.1%, with an increase of 5.8% because of CAD. Average time required, per reader, to read a case was 25 seconds without CAD and 45 seconds with CAD. CONCLUSION: CAD increases sensitivity of single reading by 1.3%, whereas double reading increases sensitivity by 8.2%. PMID- 16244253 TI - Free-response receiver operating characteristic evaluation of lossy JPEG2000 and object-based set partitioning in hierarchical trees compression of digitized mammograms. AB - PURPOSE: To assess the effects of two irreversible wavelet-based compression algorithms--Joint Photographic Experts Group (JPEG) 2000 and object-based set partitioning in hierarchical trees (SPIHT)--on the detection of clusters of microcalcifications and masses on digitized mammograms. MATERIALS AND METHODS: The use of the images in this retrospective image-collection study was approved by the institutional review board, and patient informed consent was not required. One hundred twelve mammographic images (28 with one or two clusters of microcalcifications, 19 with one mass, 17 with both abnormal findings, and 48 with normal findings) obtained in 60 women who ranged in age from 25 to 79 years were digitized and compressed at 40:1 and 80:1 by using the JPEG2000 and object based SPIHT methods. Five experienced radiologists were asked to locate and rate clusters of microcalcifications and masses on the original and compressed images in a free-response receiver operating characteristic (FROC) data acquisition paradigm. Observer performance was evaluated with the jackknife FROC method. RESULTS: The mean FROC figures of merit for detecting clusters of microcalcifications, masses, and both radiographic findings on uncompressed images were 0.80, 0.81, and 0.72, respectively. With object-based SPIHT 80:1 compression, the corresponding values were larger than the values for uncompressed images by 0.005, 0.009, and -0.005, respectively. The 95% confidence interval for the differences in figures of merit between compressed and uncompressed images was -0.039, 0.033 for the microcalcification finding; -0.055, 0.034 for the mass finding; and -0.039, 0.030 for both findings. Because each of these confidence intervals includes zero, no significant difference in detection accuracy between uncompressed and object-based SPIHT 80:1 compression was observed at a P value of 5%. The F test of the null hypothesis that all of the modes (uncompressed and four compressed modes) were equivalent yielded the following results: F = 0.255, P = .903 for the microcalcification finding; F = 0.340, P = .848 for the mass finding; and F = 0.122, P = .975 for both findings. CONCLUSION: To within the accuracy of these measurements, lossy compression of digital mammographic data at 80:1 with JPEG2000 or the object-based SPIHT algorithm can be performed without decreasing the rate of detection of clusters of microcalcifications and masses. PMID- 16244254 TI - Coregistered MR imaging myocardial viability maps and multi-detector row CT coronary angiography displays for surgical revascularization planning: initial experience. AB - PURPOSE: To evaluate assignment of left ventricular (LV) myocardial segments to coronary arterial territories by using coregistered magnetic resonance (MR) imaging and multi-detector row computed tomography (CT) displays; to assess the accuracy of coregistered displays in determining the distribution of clinically important coronary artery disease (CAD) and regional effect of CAD on LV myocardium in patients with chronic ischemic heart disease (CIHD); and to determine the utility of coregistered displays in optimizing surgical revascularization planning. MATERIALS AND METHODS: This study was HIPAA compliant and was approved by the local Institutional Review Board, with waiver of informed consent. Twenty-six patients (19 men, seven women; age, 56 years +/- 12 [+/- standard deviation]) with CIHD underwent MR imaging assessment of myocardial viability and multi-detector row CT assessment of CAD on the same day. For coregistration, a population-based LV model was fit to each data set separately; models were then registered spatially. For data analysis, correspondence between coregistered displays and the 17-segment LV model for assessment of CIHD was evaluated, accuracy of using coregistered displays to evaluate the extent of CAD and myocardial disease was assessed, and utility of coregistered displays in optimizing surgical revascularization planning was determined. RESULTS: Coronary assignment for coregistered displays and the 17-segment LV model differed in 17% of myocardial segments. For the majority of patients, three segments (midanterolateral [62%], apical lateral [73%], and apical inferior [58%]) were discordant. Segments were supplied by the left anterior descending artery, a diagonal branch, or a ramus intermedius with diagonal distribution in all but one case. Coregistered displays were deemed concordant with selective coronary angiography and alternate myocardial imaging in all cases. Overall, surgical planning was potentially enhanced in 83% of cases because, compared with alternate imaging modalities, coregistered displays were believed to demonstrate the relationship between coronary arteries and underlying myocardial tissue more definitively and efficiently (for patients in whom surgery was performed) or more correctly and comprehensively (for a presumably better-tailored surgery). CONCLUSION: Assessment of CIHD can be improved by using coregistered displays that directly relate the condition of LV myocardium to the anatomy of the coronary arteries in individual patients. PMID- 16244256 TI - Case 88: Congenital generalized lipodystrophy. PMID- 16244257 TI - Liver metastases in rats: chemoembolization combined with interstitial laser ablation for treatment. AB - PURPOSE: To assess the effect of transcatheter arterial chemoembolization (TACE) combined with laser-induced thermotherapy (LITT) for treatment of liver metastases in an animal model. MATERIALS AND METHODS: All experiments were approved by the German government and the institutional animal research review board. After subcapsular liver implantation of colorectal cancer cells in 30 WAG rats (on day 0), the animals were randomly assigned to three interventional treatment groups. In the 10 rats in group A, TACE was performed: Fourteen days after cancer cell implantation and within 20 minutes after laparotomy and retrograde placement of a microcatheter into the gastroduodenal artery, these rats were injected with mitomycin (0.1 mg), iodized oil (0.1 mL), and degradable starch microspheres (5.0 mg). In the 10 rats in group B, LITT was performed: Also on day 14, the tumors in these animals were exposed to Nd:YAG laser light of 1064 nm at 2 W for 5 minutes. In the 10 rats in group C, combined treatment was administered: TACE was performed on day 14, and LITT was performed on day 21. Tumor volumes were measured before (on day 13) and after (on day 28) treatment with magnetic resonance (MR) imaging, and the mean tumor growth ratio (day 13 tumor volume divided by day 28 tumor volume) was calculated. RESULTS: The mean tumor volumes measured before and after the treatments were, respectively, 0.11 and 0.60 cm(3) in group A, 0.11 and 0.68 cm(3) in group B, and 0.11 and 0.35 cm(3) in group C. The mean tumor growth ratio was 5.42 in group A, 6.14 in group B, and 3.15 in group C. According to Bonferroni test results, compared with the rats in groups A and B (controls), the group C rats had significantly inhibited tumor growth (P < .01 for both comparisons). CONCLUSION: Use of combined TACE LITT treatment, compared with the use of TACE or LITT alone, significantly inhibits tumor growth. PMID- 16244258 TI - Assessment of DNA damage in target tumor cells after thermoablation in mice. AB - PURPOSE: To determine the effects of temperature on cell death for cells in culture and to compare these effects with the results of in vivo experiments in which heating is induced in mice with implanted human adenocarcinoma by using magnetic methods. MATERIALS AND METHODS: Experimentation was approved by the regional animal care committee. Human adenocarcinoma cells (MX-1) and human fibroblasts (HTB-125) were exposed to defined temperatures of 45 degrees -90 degrees C for 4 minutes. Single- and double-strand DNA breaks (expressed as a percentage of the total DNA in tail) were identified by using the alkaline comet assay, and cell survival was determined by using the cloning assay and trypan blue exclusion. For in vivo experiments, MX-1 tumors were implanted into 14 mice. Magnetic heating at temperatures of 59 degrees -96 degrees C was subsequently performed by injecting magnetic material into the tumor (7 mg +/- 3 [+/- standard deviation] magnetite per tumor) and applying an alternating magnetic field (8.8 kA/m; 400 kHz) for 4 minutes. The efficiency of the temperature-dependent induction of DNA damage in isolated tumor cells was quantified and compared with that in cultured cells. RESULTS: Results of experiments with cell cultures revealed a strong correlation between DNA damage, cell survival, and temperature, as determined with the cloning assay and trypan blue exclusion. The threshold thermoablasive temperature for tumor cell elimination was found to be 55 degrees 60 degrees C. Moreover, a strong impairment in cell survival was found when damaged DNA accounted for more than 50% of the total DNA. The heating sensitivities of malignant and nonmalignant cells did not differ. After the magnetic heating of tumors in vivo to temperatures of up to 96 degrees C (rectal temperatures between 27 degrees C +/- 2 and 29 degrees C +/- 2), isolated tumor cells showed a mean of 71.9% +/- 24.5 of total DNA in the tail per cell compared with nontreated tumors, which showed 8.0% +/- 3.1. CONCLUSION: There appears to be a threshold temperature for the induction of irreversible DNA damage. This is reflected by the results of in vivo experiments in tumor-bearing mice after high temperatures were applied to tumors by using magnetic heating. PMID- 16244259 TI - Accuracy of liver fat quantification at MR imaging: comparison of out-of-phase gradient-echo and fat-saturated fast spin-echo techniques--initial experience. AB - PURPOSE: To retrospectively determine the relative accuracy of liver fat quantification with out-of-phase gradient-echo magnetic resonance (MR) imaging and fat-saturated fast spin-echo MR imaging in patients with and without cirrhosis, with histologic analysis as the reference standard. MATERIALS AND METHODS: Committee on Human Research approval was obtained. Patient consent was not required. Data collection ended before HIPAA regulations were implemented, but patient anonymity was maintained. Twenty-seven patients, 16 with cirrhosis, were retrospectively identified who underwent MR imaging before histopathologic evaluation of liver fat at biopsy or surgery. The patient population consisted of 15 male and 12 female patients (mean age, 55 years; range, 16-75 years). One radiologist blinded to the histopathologic results recorded mean signal intensity derived from three regions of interest placed in the right and left lobes of the liver on three sections and signal intensity of the spleen from one region of interest within the same section. Liver fat was quantified with the relative loss of signal intensity on out-of-phase images compared with that on in-phase T1 weighted gradient-echo images and with relative loss of signal intensity on T2 weighted fast spin-echo MR images obtained with fat saturation compared with those obtained without fat saturation. Hotelling t test was used to compare correlation coefficients between relative signal intensity differences and histopathologically determined percentage of fat. RESULTS: In patients without cirrhosis, liver fat quantification with fat-saturated fast spin-echo MR imaging was significantly better than it was with out-of-phase gradient-echo MR imaging (r = 0.92 vs 0.69, P < .01). In patients with cirrhosis, liver fat quantification was correlated only with fat-saturated fast spin-echo MR imaging (r = 0.76, P < .01); the relative signal intensity loss on out-of-phase gradient-echo MR images was not correlated with histopathologically determined percentage of fat (r = 0.25, P = .36). CONCLUSION: Preliminary results suggest liver fat may be more accurately quantified with fat-saturated fast spin-echo MR imaging than with out of-phase gradient-echo MR imaging, especially in patients with cirrhosis. PMID- 16244260 TI - Cirrhotic nodules: association between MR imaging signal intensity and intranodular blood supply. AB - PURPOSE: To retrospectively determine whether there is a relationship between the intranodular blood supply evaluated at computed tomography (CT) during arterial portography (CTAP) and CT during hepatic arteriography (CTHA) and the magnetic resonance (MR) imaging signal intensity of nodules associated with cirrhosis. MATERIALS AND METHODS: Neither institutional review board approval nor informed consent was required for retrospective reviews of medical records and images. One hundred fourteen hepatocellular nodules 10 mm or greater in largest diameter in 58 patients (39 men, 19 women; mean age, 61 years) with cirrhosis were evaluated at CTAP, CTHA, and MR imaging. The CTAP and CTHA nodule findings were divided into three main types: Type A nodules were isoattenuating at CTAP and hypoattenuating at CTHA; type B nodules, slightly hypoattenuating at CTAP and hypoattenuating at CTHA; and type C nodules, strongly hypoattenuating at CTAP and hyperattenuating at CTHA. The relationships between the CTAP and CTHA findings and the MR imaging signal intensity among these nodules were analyzed by using the chi(2) test. RESULTS: On T1-weighted MR images, 27 (63%) of 43 type A nodules were hyperintense, nine (39%) of 23 type B nodules were isointense, and 19 (48%) of 40 type C nodules were hypointense; differences were not significant. On T2 weighted MR images, 31 (72%) of 43 type A nodules were hypointense (P < .05), 12 (52%) of 23 type B nodules were isointense, and 34 (85%) of 40 type C nodules were hyperintense (P < .05). CONCLUSION: There was a significant association between intranodular blood supply and nodule signal intensity on T2-weighted MR images. However, study findings did not show whether the blood itself (ie, blood volume or blood flow amount) directly influenced the signal intensity. PMID- 16244261 TI - Neoangiogenesis and sinusoidal capillarization in hepatocellular carcinoma: correlation between dynamic CT and density of tumor microvessels. AB - PURPOSE: To retrospectively evaluate the correlation between the degree of contrast enhancement on dynamic computed tomographic (CT) scans and the degree of neoangiogenesis and sinusoidal capillarization in hepatocellular carcinoma (HCC). MATERIALS AND METHODS: The institutional review board did not require approval or informed patient's consent for the review of medical records or images. Dynamic CT scans of 97 nodular HCCs in 97 patients (79 men, 18 women; age range, 29-73 years; mean age, 54 years) were evaluated in terms of the attenuation change in the arterial, portal venous, and delayed phases, and the results were correlated with the number of unpaired arteries and the degree of sinusoidal capillarization at histopathologic examination. The mean attenuation value of the nodular HCCs on triple-phase helical CT scans was correlated with the number of unpaired arteries and the degree of sinusoidal capillarization. Statistical analysis was performed with the Spearman rank correlation test. RESULTS: The number of unpaired arteries in the nodular HCCs was found to correlate with the degree of contrast enhancement in the arterial phase (r = 0.225, P = .027), but did not correlate with the degree of contrast enhancement in the portal and delayed phases. The degree of sinusoidal capillarization did not correlate linearly with the mean attenuation of the nodular HCCs in any phase of contrast enhancement. CONCLUSION: The degree of contrast enhancement of the nodular HCCs in the arterial phase tended to correlate with the number of unpaired arteries, but no correlation was evident between the degree of contrast enhancement and sinusoidal capillarization in any phase of CT imaging. PMID- 16244262 TI - Esophageal varices: evaluation with esophagography with barium versus endoscopic gastroduodenoscopy in patients with compensated cirrhosis--blinded prospective study. AB - PURPOSE: To prospectively evaluate the accuracy of esophagography with barium in diagnosis of esophageal varices (EV) in patients with compensated cirrhosis, with endoscopic gastroduodenoscopy as the reference standard. MATERIALS AND METHODS: In this study, which was approved by the local Helsinki Committee and in which all patients consented to participate, 61 patients with cirrhosis (34 men, 27 women; mean age, 61 years; range, 36-76 years) received a diagnosis clinically or with liver biopsy. In 87% (n = 53) of patients, Child-Pugh classification was A; in 13% (n = 8), Child-Pugh classification was B. They were evaluated with endoscopic gastroduodenoscopy, according to Japanese general criteria. Esophagography was performed within 3 weeks of endoscopic gastroduodenoscopy, and EV were assigned grades as follows: 0, no EV were seen; 1, EV manifested as very mild irregularities of the folds; and 2, the irregularity of the folds (EV) was clearly present. They were also assigned grades for shape and size: grade F0, no EV detected; grade F1, small straight EV; grade F2, slightly enlarged tortuous EV occupying less than one-third of esophageal lumen; and grade F3, large coil shaped EV that occupied more than one-third of esophageal lumen. The sensitivity and specificity and positive and negative predictive values of esophagography for identification of each grade of EV were calculated separately, as was the 95% confidence interval. RESULTS: All large EV (grades F2 and F3) were diagnosed at esophagography. Sensitivity declined with small EV (grade F1) to 71. The overall sensitivity of esophagography was 89% (95% confidence interval: 75.9%, 96.5%), the overall specificity was 83% (95% confidence interval: 64.5%, 94.7%), the overall positive predictive value was 89%, and the overall negative predictive value was 83% (95% confidence interval: 64.5%, 94.7%). Overall accuracy was 87%. CONCLUSION: Esophagography is highly accurate in diagnosis of EV and can be considered a viable noninvasive alternative for determination of patients who should be selected for prophylactic treatment. PMID- 16244263 TI - Staging prostate cancer with dynamic contrast-enhanced endorectal MR imaging prior to radical prostatectomy: experienced versus less experienced readers. AB - PURPOSE: To prospectively determine the accuracy of experienced and less experienced readers in the interpretation of combined T2-weighted fast spin-echo (SE) magnetic resonance (MR) images and dynamic contrast material-enhanced MR images compared with T2-weighted fast SE alone, with respect to differentiation of stage T2 versus stage T3 prostate carcinoma, with histologic analysis serving as the reference standard. MATERIALS AND METHODS: Institutional review board approval and informed consent were obtained, and 124 consecutive men (age range, 42-74 years; median age, 63 years) with biopsy-proved prostate cancer underwent MR imaging and were candidates for radical prostatectomy. T2-weighted fast SE MR images and multisection dynamic contrast-enhanced MR images with a 2-second time resolution for the whole prostate were obtained. The T2-weighted and fused color coded parametric dynamic contrast-enhanced MR images with T2-weighted images were evaluated prospectively and scored with regard to local extent by one experienced reader and evaluated retrospectively by two less experienced readers working in consensus by using a five-point scale; images with a score greater than or equal to four were considered indicative of T3 disease. Results were correlated with whole-mount section histopathologic findings, and receiver operating characteristics analysis was performed. RESULTS: Twenty-five patients were excluded because of positive findings in the lymph nodes (n = 16), preoperative biopsy-proved seminal vesicle invasion (n = 5), and an absent dynamic dataset (n = 4). Ninety-nine patients were included in this study. The overall sensitivity, specificity, and accuracy of MR staging performance in prostate cancer with dynamic contrast-enhanced MR imaging was 69% (24 of 35 patients), 97% (62 of 64 patients), and 87% (86 of 99 patients), respectively, for the experienced reader. This difference was not significant (P = .48) when results were compared with results from the T2-weighted images. Staging performance for the less experienced readers with parametric dynamic contrast-enhanced MR imaging, however, resulted in significant improvement of the area under the receiver operating characteristics curve (A(z)) compared with T2-weighted MR imaging alone (A(z) = .66 and .82, respectively; P = .01). CONCLUSION: The use of multisection dynamic contrast-enhanced MR imaging in staging prostate cancer showed significant improvement in staging performance for the less experienced readers but had no benefit for the experienced reader. PMID- 16244264 TI - Testicular microlithiasis in association with pseudoxanthoma elasticum. AB - PURPOSE: To determine the presence of testicular microlithiasis in male subjects with pseudoxanthoma elasticum (PXE). MATERIALS AND METHODS: Institutional review board approval was obtained for the prospective and retrospective components of this HIPAA-compliant study. Informed consent was obtained from all patients or their parents. Testicular ultrasonography (US) was performed in eight men aged 29 56 years and in one 13-year-old boy, all with confirmed PXE. Two radiologists reviewed the US images by consensus for testicular microlithiasis, testicular masses, and additional testicular abnormalities. Testicular microlithiasis was judged to be classic when at least five microliths were seen on a single US image and to be limited when fewer than five microliths were seen on all obtained US images. Urologic physiologic examinations were performed. A history and/or symptoms of testicular disease also were recorded at the time of examination. Similarly, the testicular US images obtained in two additional men, aged 48 and 59 years, and in another 13-year-old boy were retrospectively reviewed. Histopathologic testicular analysis was performed in one autopsy case. RESULTS: Of the 12 participants, 11 (92%) had classic and one (8%) had limited testicular microlithiasis. None of the 12 participants had evidence of testicular malignancy at US or physical examination. Histopathologic analysis at autopsy revealed intratubular microlithiasis without the calcification of elastic fibers in arterial walls that is characteristic of cutaneous PXE. CONCLUSION: Study findings suggested an association between PXE and testicular microlithiasis. It is possible that the testicular microlithiasis in male subjects who have PXE is related to the underlying PXE abnormality. PMID- 16244265 TI - Salivary gland tumors at in vivo proton MR spectroscopy. AB - PURPOSE: To prospectively evaluate whether proton magnetic resonance (MR) spectroscopy can be used to characterize salivary gland tumors (SGTs). MATERIALS AND METHODS: Ethics committee approval and informed consent were obtained. Hydrogen 1 ((1)H) MR spectroscopy was performed with echo times of 136 and 272 msec at 1.5 T in both SGTs and normal parotid glands. Spectra were analyzed in the time domain by using prior knowledge in the fitting procedure to obtain peak amplitudes of choline (Cho), creatine (Cr), and unsuppressed water. Mean Cho/Cr and Cho/water ratios for each subgroup of SGTs were obtained, and results were compared by using a nonparametric t test. RESULTS: Successful spectra were acquired in 56 patients (35 men, 21 women; mean age, 56 years) with a total of nine malignant tumors and 47 benign SGTs (24 Warthin tumors, 22 pleomorphic adenomas, one oncocytoma). At an echo time of 136 msec, Cho/Cr ratios were obtained in 26 (47%) of 55 spectra, with a mean value (+/- standard deviation) of 1.73 +/- 0.47, 5.49 +/- 1.86, 3.46 +/- 0.84, and 2.45 for malignant tumors, Warthin tumors, pleomorphic adenomas, and oncocytoma, respectively. Differences were significant between Warthin tumors and pleomorphic adenomas (P = .028) and between benign SGTs and malignant tumors (P < .001). At an echo time of 272 msec, Cho/Cr ratios were obtained in 16 (30%) of 53 spectra, with a mean value of 2.27 +/- 0.69, 6.92 +/- 1.47, and 3.67 +/- 1.23 for malignant tumors, Warthin tumors, and pleomorphic adenomas, respectively. Differences were also significant between Warthin tumors and pleomorphic adenomas (P = .041) and benign SGTs and malignant tumors (P = .004). There was a significant difference in mean Cho/water ratio for Warthin tumors versus pleomorphic adenomas at echo times of 136 msec (P = .003) and 272 msec (P = .002) but not for benign SGTs versus malignant tumors. CONCLUSION: (1)H MR spectroscopy may be used to characterize SGTs, but a larger study is required to validate these initial results. PMID- 16244266 TI - Orthopedic hardware complications diagnosed with multi-detector row CT. AB - PURPOSE: To retrospectively evaluate multi-detector row computed tomography (CT) for the depiction of orthopedic hardware complications in the spine and appendicular skeleton. MATERIALS AND METHODS: This HIPAA-compliant study had institutional review board approval; patient informed consent was not required. Results of 114 multi-detector row CT studies performed because of possible hardware complications in 109 patients (57 men, 52 women; mean age, 44 years; age range, 12-82 years) were available for analysis. The CT studies were retrospectively reviewed and compared with clinical or surgical outcomes, which were used as the reference standard. In another experiment, detection of hardware complications on radiographs and multi-detector row CT images was compared between two readers for selected cases (18 positive and 26 negative) by using receiver operating characteristic (ROC) methods. RESULTS: For 91 (80%) of 114 multi-detector row CT studies, the complication status could be determined on the basis of clinical or surgical outcomes. Twenty-three multi-detector row CT studies were confirmed to be positive (revealing 10 cases of nonunion, five cases of hardware malplacement, three cases of hardware loosening, three perihardware fractures, and two chronic infections), and 57 were confirmed to be negative. There were three false-positive and eight false-negative multi-detector row CT studies. With clinical or surgical outcomes as the reference standard, the sensitivity, specificity, and positive and negative predictive values of multi detector row CT were 74% (23 of 31 studies), 95% (57 of 60 studies), 88% (23 of 26 studies), and 88% (57 of 65 studies), respectively. Results of ROC analysis indicated that detection of hardware complications was much lower with radiography than with multi-detector row CT (area under ROC curve, 0.84 vs 1.00; F = 4.69, df = 1, 43; P < .05). CONCLUSION: Multi-detector row CT is an effective tool for depicting orthopedic hardware complications. PMID- 16244267 TI - Anterior shoulder instability: accuracy of MR arthrography in the classification of anteroinferior labroligamentous injuries. AB - PURPOSE: To retrospectively evaluate the accuracy of magnetic resonance (MR) arthrography in the classification of anteroinferior labroligamentous injuries by using arthroscopy as the reference standard. MATERIALS AND METHODS: Ethical committee approval and informed consent were obtained. MR arthrograms obtained in 205 patients, including a study group of 104 patients (74 male and 30 female; mean age, 28.2 years) with arthroscopically proved labroligamentous injuries and a control group of 101 patients (65 male and 36 female; mean age, 31.4 years) with intact labroligamentous complex, were reviewed in random order. MR arthrograms were analyzed for the presence and type (Bankart, anterior labral periosteal sleeve avulsion [ALPSA], Perthes, glenolabral articular disruption [GLAD], or nonclassifiable lesion) of labroligamentous injuries by two radiologists in consensus. Results were compared with arthroscopic findings. Sensitivity, specificity, accuracy, and corresponding 95% confidence intervals for the detection and classification of anteroinferior labroligamentous lesions with MR arthrography were calculated. RESULTS: At arthroscopy, 104 anteroinferior labroligamentous lesions were diagnosed, including 44 Bankart lesions, 22 ALPSA lesions, 12 Perthes lesions, and three GLAD lesions. Twenty-three labral lesions were nonclassifiable at arthroscopy, all of which occurred after a history of chronic instability. Nineteen (83%) of these 23 lesions were also nonclassifiable at MR arthrography. With arthroscopy used as the reference standard, labroligamentous lesions were detected and correctly classified at MR arthrography with sensitivities of 88% and 77%, specificities of 91% and 91%, and accuracies of 89% and 84%, respectively. Bankart, ALPSA, and Perthes lesions were correctly classified in 80%, 77%, and 50% of cases, respectively. The three GLAD lesions were all correctly assessed. CONCLUSION: MR arthrography is accurate in enabling classification of acute and chronic anteroinferior labroligamentous injuries, although correct interpretation of Perthes lesions remains difficult. PMID- 16244268 TI - Benign and malignant processes: normal values and differentiation with chemical shift MR imaging in vertebral marrow. AB - PURPOSE: To establish retrospectively a range of values for signal intensity change in normal vertebral marrow by using chemical shift magnetic resonance (MR) imaging and to assess the use of this technique in differentiating benign from malignant marrow abnormalities. MATERIALS AND METHODS: Institutional Review Board approval for this retrospective, HIPAA-compliant study was obtained; informed consent was waived. A total of 569 normal vertebrae in 75 patients (42 women, 33 men; mean age, 57.5 years; age range, 26-84 years) (control group) and 221 lesions in 92 patients (50 women, 42 men; mean age, 59.0 years; age range, 27-85 years) (study group) who had focal vertebral marrow abnormalities were studied by using 1.5-T chemical shift MR imaging. Imaging time was less than 1 minute. The proportional change in signal intensity on in-phase compared with out-of-phase images was calculated by using 1 x 1-cm regions of interest (ROIs) in the control group and ROIs as large as possible for focal lesions in the study group. This change in signal intensity (expressed as a percentage) was compared with that of normal levels and benign and malignant lesions. For statistical analysis, a random effect model was used that was adjusted for multiple comparisons. RESULTS: A substantial decrease in signal intensity was noted for all normal vertebrae (mean, 58.5%) and for benign lesions, including endplate degeneration (mean, 52.2%), Schmorl nodes with edema (mean, 58.0%), hemangiomas (mean, 49.4%), and benign fractures (mean, 49.3%). Metastases exhibited either a minimal decrease or an increase in signal intensity (mean, 2.8%). Although there was some overlap in the range of signal intensity values among malignant lesions, benign lesions, and normal marrow, the differences in signal intensity loss for normal marrow and benign and malignant lesions were significant (P < .01 for all pairwise comparisons after adjusting for multiplicity). CONCLUSION: Bone marrow in the vertebral bodies displays somewhat variable behavior at chemical shift MR imaging. Results suggest that a decrease in signal intensity greater than 20% on out-of-phase images compared with in-phase images should be used as a cutoff threshold for normalcy to allow distinction between benign and malignant causes of vertebral marrow abnormalities. PMID- 16244269 TI - Acute low back pain and radiculopathy: MR imaging findings and their prognostic role and effect on outcome. AB - PURPOSE: To prospectively determine in patients with acute low back pain (LBP) or radiculopathy, the magnetic resonance (MR) imaging findings, prognostic role of these findings, and effect of diagnostic information on outcome. MATERIALS AND METHODS: Institutional review board approval and informed consent were obtained. This study was HIPAA compliant. A total of 246 patients with acute-onset LBP or radiculopathy were randomized to either the early information arm of the study, with MR results provided within 48 hours, or the second arm of the study, where both patients and physicians were blinded to MR results, unless this information was critical to patient care. Patients underwent 6 weeks of conservative care. Roland function scoring, visual pain analog, Short Form 36 health status survey, self-efficacy scoring, and a fear avoidance questionnaire were completed at presentation; at 2-, 4-, 6-, and 8-week follow-up; and at 6-, 12-, and 24-month follow-up. A second MR imaging examination was performed at 6-week follow-up. Multivariate logistic regression analysis was used to determine which imaging and nonimaging variables can be used to predict improvement in Roland function and patient satisfaction. The chi(2) test and repeated-measures analysis of variance were used to compare outcome of blinded and unblinded patients. RESULTS: Herniation was identified in 60% (n = 147) of patients at the initial examination. The prevalence of herniations in patients with LBP (57%) (n = 85) and those with radiculopathy (65%) (n = 62) were similar (P = .217), although patients with radiculopathy were more likely to have stenosis and nerve root compression (P < .006). There was no relationship between herniation type, size, and behavior over time with outcome. An improvement of 50% or more in Roland function score at 6-week follow-up occurred 2.7 times as often among patients with a herniation at baseline (P = .003). Improvement at 6-week follow-up was similar in unblinded (60%) (n = 55) and blinded (67%) (n = 57) patients (P = .397). Self-efficacy, fear avoidance beliefs, and the Short Form 36 subscales were similar for blinded and unblinded patients. CONCLUSION: In typical patients with LBP or radiculopathy, MR imaging does not appear to have measurable value in terms of planning conservative care. Patient knowledge of imaging findings does not alter outcome and is associated with a lesser sense of well-being. PMID- 16244270 TI - Endovascular treatment with coils of 149 middle cerebral artery berry aneurysms. AB - PURPOSE: To retrospectively evaluate the immediate and long-term clinical results, as well as the angiographic results, of occlusion of middle cerebral artery (MCA) berry aneurysms with coils. MATERIALS AND METHODS: This retrospective study had institutional review board approval, and informed consent was obtained. One hundred fifty-four MCA aneurysms in 142 patients were intended to be treated. Complications, patient clinical outcomes, and immediate postprocedural and follow-up angiography results were retrospectively evaluated. RESULTS: One hundred forty-nine (96.8%) of 154 MCA aneurysms (72 ruptured, 77 unruptured) were occluded with coils in 137 patients (99 women and 38 men; age range, 28-76 years; mean, 48 years). Thromboembolic events occurred in 20 (13.4%) and aneurysm perforation occurred in seven (4.7%) of 149 procedures. Endovascular treatment (EVT) was performed without complications for 121 (81.2%) of the treated aneurysms. For ruptured aneurysms, the treatment-related mortality rate was 6% (four of 72 aneurysms) and the treatment-induced permanent morbidity rate was 1% (one aneurysm). For unruptured aneurysms, the treatment-induced mortality rate was 1% (one of 77 aneurysms) and the procedure-related permanent morbidity rate was 3% (two aneurysms). One hundred five (70.5%) of the 149 aneurysms were examined with follow-up angiography at least once. Recurrences were found for 21 (20%) of the 105 aneurysms that were followed up for a cumulative period of 1564 months (mean, 15 months). Of these 21 recurrent aneurysms, 10 increased in size in the interval between follow-up angiography examinations and 11 remained stable. A second treatment was required for 12 aneurysms, and a third treatment was required for one. After repeat EVT, total aneurysm occlusion was attained for nine aneurysms, and a residual neck was seen in two aneurysms. One recurrent aneurysm was surgically clipped. The nine other aneurysms with small recurrences were not candidates for additional treatment. CONCLUSION: EVT of MCA aneurysms with coils can be successfully performed without inducing neurologic deficits in most patients with ruptured or unruptured aneurysms. PMID- 16244271 TI - CT appearance of bone metastases detected with FDG PET as part of the same PET/CT examination. AB - PURPOSE: To retrospectively evaluate lesion findings at computed tomography (CT) performed as part of a combined positron emission tomography (PET)/CT examination in patients suspected of having metastatic bone lesions-lesions that were detected with fluorine 18 fluorodeoxyglucose (FDG) PET as part of the same examination-and to correlate the CT and FDG PET findings. MATERIALS AND METHODS: This HIPAA-compliant study had institutional review board approval, and the need for patient informed consent was waived. Three hundred fifty-nine consecutive patients (191 male patients, 168 female patients; mean age, 56.9 years; age range, 8-92 years) underwent PET/CT. PET images were first reviewed by nuclear medicine physicians who had no clinical information regarding the presence or absence of bone metastasis by using a five-point grading system (0, a lesion was definitely negative for metastasis; 1, a lesion was probably negative; 2, a lesion was equivocal; 3, a lesion was probably positive; and 4, a lesion was definitely positive). For lesions assigned a grade of 3 or 4 at PET, CT characteristics such as the presence or absence of morphologic changes or accompanying findings (including bone destruction) were assessed by radiologists on the CT images obtained during the same imaging session. RESULTS: One hundred seventy-nine lesions in 55 patients were considered to be probable or definite bone metastases at PET. One hundred thirty-three of these lesions in 33 patients were clinically confirmed to be bone metastases at follow-up and/or histopathologic examination. CT revealed osteolytic changes in 41 (31%) and osteoblastic changes in 21 (16%) of the 133 lesions, but no or nonspecific changes were seen at CT in 49 (37%) and 22 lesions (17%), respectively. Of the 179 lesions suspected at PET, 46 ultimately proved to be nonosseous or false positive for bone metastasis. Of these 46 lesions, 38 were not located in the bone but in adjacent tissues such as the pleura. CONCLUSION: CT images obtained as part of PET/CT scanning were useful in yielding the precise location of bone lesions and thus helping avoid misdiagnosis of bone metastasis; however, CT revealed morphologic changes in only half of the lesions assigned a grade of 3 or 4 at PET. PMID- 16244272 TI - Comparison of alignment of computer-registered data sets: combined PET/CT versus independent PET and CT of the thorax. AB - PURPOSE: To retrospectively determine whether alignment of registered positron emission tomographic (PET) and computed tomographic (CT) data sets obtained independently varies significantly from alignment of data sets acquired from a combined PET/CT scanner. MATERIALS AND METHODS: The study was approved by the institution's Human Research Committee with a waiver of informed consent and complied with HIPAA. Whole-body combined PET/CT data sets and separate routinely positioned thoracic CT data sets were obtained from 12 patients (six men, six women; mean age, 48.6 years; range, 24-62 years). Separate PET and thoracic CT data sets matched for patient positioning and respiration were acquired on the same day for nine patients (four men, five women; mean age, 71 years; range, 51 90 years). Computer nonlinear registration was performed on PET and CT data sets from combined PET/CT (fusion group 1), PET data sets from combined PET/CT with unmatched thoracic CT (fusion group 2), and data sets from separate PET and CT matched for patient positioning and respiration (fusion group 3). Quality of alignment was assessed by two radiologists in consensus blinded to the source of registered data in each fusion group at the following anatomic locations: diaphragm, aortic arch, heart, thoracic spine, and lung apices. Results were compared by using the Wilcoxon paired signed rank and unpaired rank sum tests. RESULTS: Quality of alignment did not significantly differ between fusion group 1 and fusion group 3. Fusion group 1 provided significantly better alignment in two of five anatomic locations (P = .008 for diaphragm and P = .031 for heart) than fusion group 2. Fusion group 3 provided significantly better alignment in two of five anatomic locations (P = .037 for diaphragm and P = .009 for heart) than fusion group 2. CONCLUSION: Thoracic anatomic alignment does not significantly differ between registered PET and CT data sets acquired on a combined PET/CT scanner or from separate PET and CT scanners obtained on the same day when carefully matched for anatomic positioning and respiration. PMID- 16244273 TI - The focal hepatic hot spot sign. PMID- 16244274 TI - Sixteen-detector row CT angiography for lower-leg arterial occlusive disease: analysis of section width. AB - Institutional review board approval and written informed consent from all patients were obtained. Diagnostic accuracy of three reconstructions of 16 detector row computed tomographic (CT) angiography data with different section widths and increments (2.0 and 1.0 mm [CT data set 1], 1.0 and 0.5 mm [CT data set 2], and 0.75 and 0.4 mm [CT data set 3]) was compared with that of digital subtraction angiography (DSA) in 163 arterial segments in 17 patients with occlusive peripheral arterial disease (PAD). Arterial visibility was superior with CT as compared with DSA (P < .008). Sensitivity for stenosis detection did not differ between the CT reconstructions, whereas specificity was significantly improved when CT data set 3 was used (P < .017). Stenosis length did not differ significantly between CT angiography and DSA. Accuracy of stenosis detection was 88.2%, 90.8%, and 96.1% with CT data sets 1, 2, and 3, respectively. CT angiography has excellent diagnostic accuracy in the assessment of lower-leg PAD provided that the thinnest possible section width is used. PMID- 16244275 TI - Small-bowel perfusion measurement: feasibility with single-compartment kinetic model applied to dynamic contrast-enhanced CT. AB - This study was institutional review board approved and HIPAA compliant. Informed consent was obtained from all patients. The purpose of the study was to prospectively examine the feasibility of measuring small-bowel quantitative blood flow by using motion-corrected, contrast-enhanced computed tomographic (CT) images and a single-compartment kinetic model. Seven patients underwent abdominal CT in which 40 10-mm-thick sections were obtained at a single level. Small-bowel images were obtained every 3 seconds after contrast agent administration. Automated application of regions of interest yielded time-enhancement curves for the bowel wall and the aorta. A one-compartment model was applied to each set of time-enhancement curves for determination of the small-bowel volumetric blood flow F(V), volume of distribution V(D), and blood transit time tau. F(V) was also calculated by using the first-pass method and gamma variate analysis for model validation. The F(V) values obtained by using the single-compartment model (mean F(V), 0.47 min(-1)) showed excellent linear correlation with those obtained by using the first-pass method (Pearson r = 0.80) and gamma variate analysis (Pearson r = 0.97). Mean V(D) and tau values were 2.86 (unitless) and 4.28 seconds, respectively. A one-compartment kinetic model can be applied to motion corrected, contrast-enhanced small-bowel CT images to quantify perfusion. PMID- 16244276 TI - Solitary pulmonary nodule: characterization with combined wash-in and washout features at dynamic multi-detector row CT. AB - PURPOSE: To prospectively assess the accuracy of combined wash-in and washout characteristics at dynamic contrast material-enhanced multi-detector row computed tomography (CT) in distinguishing benign from malignant solitary pulmonary nodules. MATERIALS AND METHODS: Institutional review board approval and informed consent were obtained. The study included 107 patients (62 men, 45 women; mean age, 55 years; range, 22-81 years) with a solitary pulmonary nodule. After unenhanced CT (2.5-mm collimation) scans were obtained, dynamic CT was performed by using a helical technique (series of images obtained throughout the nodule, with 2.5-mm collimation, at 30, 60, 90, and 120 seconds and 4, 5, 9, 12, and 15 minutes) after intravenous injection of contrast medium (120 mL). Tissue diagnosis was made in 70 nodules, and follow-up images showed benignancy in the remaining 37 (no change in size, n = 32; decrease in size, n = 5). CT findings were analyzed in terms of wash-in and washout of contrast medium. Sensitivity, specificity, and accuracy for malignant nodules were calculated by considering both the wash-in and washout characteristics at dynamic CT. RESULTS: There were 49 malignant and 58 benign nodules. When diagnostic criteria for malignancy of both wash-in of 25 HU or greater and washout of 5-31 HU were applied, sensitivity, specificity, and accuracy for malignancy were 94% (46 of 49 nodules), 90% (52 of 58 nodules), and 92% (98 of 107 nodules), respectively. Of 58 benign nodules, 27 showed less than 25 HU wash-in, 14 showed persistent contrast enhancement without washout and with wash-in of 25 HU or greater, and 11 showed washout greater than 31 HU and wash-in of 25 HU or greater. CONCLUSION: Evaluation of solitary pulmonary nodules by analyzing combined wash-in and washout characteristics at dynamic contrast-enhanced multi-detector row CT showed 92% accuracy for distinguishing benign nodules from malignant nodules. PMID- 16244277 TI - Computer-aided detection of peripheral lung cancers missed at CT: ROC analyses without and with localization. AB - PURPOSE: To retrospectively evaluate whether a difference-image computer-aided detection (CAD) scheme can help radiologists detect peripheral lung cancers missed at low-dose computed tomography (CT). MATERIALS AND METHODS: Institutional review board approval and informed patient and observer consent were obtained. Seventeen patients (eight men and nine women; mean age, 60 years) with a missed peripheral lung cancer and 10 control subjects (five men and five women; mean age, 63 years) without cancer at low-dose CT were included in an observer study. Fourteen radiologists were divided into two groups on the basis of different image display formats: Six radiologists (group 1) reviewed CT scans with a multiformat display, and eight radiologists (group 2) reviewed images with a "stacked" cine-mode display. The radiologists, first without and then with the CAD scheme, indicated their confidence level regarding the presence (or absence) of cancer and the most likely position of a lesion on each CT scan. Receiver operating characteristic (ROC) curves were calculated without and with localization to evaluate the observers' performance. RESULTS: With the CAD scheme, the average area under the ROC curve improved from 0.763 to 0.854 for all radiologists (P = .002), from 0.757 to 0.862 for group 1 (P = .04), and from 0.768 to 0.848 for group 2 (P = .01). The average sensitivity in the detection of 17 cancers improved from 52% (124 of 238 observations) to 68% (163 of 238 observations) for all radiologists (P < .001), from 49% (50 of 102 observations) to 71% (72 of 102 observations) for group 1 (P = .02), and from 54% (74 of 136 observations) to 67% (91 of 136 observations) for group 2 (P = .006). The localization ROC curve also improved. CONCLUSION: Lung cancers missed at low-dose CT were very difficult to detect, even in an observer study. The use of CAD, however, can improve radiologists' performance in the detection of these subtle cancers. PMID- 16244278 TI - CT fluoroscopy-guided biopsy of the lung or upper abdomen with a breath-hold monitoring and feedback system: a prospective randomized controlled clinical trial. AB - PURPOSE: To prospectively determine the clinical effectiveness of a breath-hold monitoring and feedback system in computed tomographic (CT) fluoroscopy-guided biopsies in which respiratory motion is a problem. MATERIALS AND METHODS: Institutional review board approval and oral and written informed consent were obtained. This study was HIPAA compliant. A bellows-based system was used to monitor respiration and provide patient feedback. A randomized controlled clinical trial compared intermittent mode CT fluoroscopy-guided biopsies of the lung or upper abdomen performed with (n = 56) and without (n = 57) the bellows system. Inclusion criteria for 113 patients were lesions 6 cm or smaller in maximum dimension that were not affixed to the chest or abdominal wall. Primary outcome measurements were CT fluoroscopy exposure time and patient dose. Wilcoxon rank sum, chi(2), and Fisher exact tests were used for statistical analysis. RESULTS: Median CT fluoroscopy exposure time was 12.6 seconds (range, 2.4-44.4 seconds) for the bellows group and 18.0 seconds (range, 6.0-118.0 seconds) for the nonbellows group (P = .004). Patient dose was decreased in the bellows group (median dose, 29.5 mGy; range, 4.7-135.8 mGy) versus the nonbellows group (median, 41.3 mGy; range, 11.8-155.9 mGy) (P = .01). Lesions were accessed successfully with one needle puncture attempt in 43 of 56 patients (77%) in the bellows group and 30 of 57 patients (53%) in the nonbellows group (P = .007). Pneumothorax developed in 11 of 50 patients (22%) in the bellows group who underwent lung biopsy compared with 16 of 50 (32%) patients in the nonbellows group. CONCLUSION: A breath-hold monitoring and feedback system allows depiction of mobile target lesions throughout CT fluoroscopy-guided biopsy of the lung and upper abdomen. PMID- 16244279 TI - Radiofrequency ablation of liver tumors: a new cause of benign portal venous gas. AB - PURPOSE: To retrospectively describe and categorize the presence of portal venous gas (PVG) from radiofrequency (RF) ablation of hepatic tumors. MATERIALS AND METHODS: The study was HIPAA compliant, and informed consent was waived. Thirty four consecutive computed tomography (CT)-guided percutaneous RF ablations of liver tumors in 26 patients (13 men, 13 women; mean age, 69 years) with five hepatocellular carcinomas and 21 metastatic liver tumors (13 colon, five other, and three unknown primary tumors) were performed with an institutional review board-approved protocol. Two treatment modalities were used: RF ablation alone (13 procedures) and combined RF ablation and ethanol injection (21 procedures). Presence of PVG was quantified with three parameters: maximum length of a portal venous branch with gas, number of Couinaud segments in which PVG was seen, and total number of portal venous branch points with gas. Then an overall PVG score from 0 to 5 was determined. Also, when tumoral gas was seen on CT scans, the largest cross-sectional area of gas was measured. The two ablation methods were compared for quantities of PVG and tumoral gas. The role of N(2)O anesthetic in PVG and tumoral gas formation during ablation also was studied. Statistical analyses were performed with Wilcoxon rank sum and Student t tests. RESULTS: In 25 procedures (74%), gas was found in portal vein branches; in 30 procedures (88%), gas was also found in tumoral and peritumoral tissues. There was no significant difference in frequency of PVG between the ablation methods. Combined therapy yielded significantly greater lengths of PVG (P < .002) and more portal venous branch points (P < .001) than did RF ablation alone. Mean PVG score was 2.4 +/- 0.4 (standard error of the mean) for combined therapy and 0.9 +/- 0.2 for RF ablation alone (P < .004). N(2)O anesthetic was associated with greater amounts of tumoral gas (P < .008) and PVG (P < .03). Tumoral gas, peritumoral gas, and PVG dissipated within 20 minutes after ablation in all patients. No morbidity or mortality was associated with PVG. CONCLUSION: RF ablation is a common yet benign cause of transient PVG, tumoral gas, and peritumoral gas. Combined RF and ethanol ablation was associated with more PVG than was RF ablation alone. PMID- 16244280 TI - DSA versus multi-detector row CT angiography in peripheral arterial disease: randomized controlled trial. AB - PURPOSE: To prospectively compare therapeutic confidence in, patient outcomes (in terms of quality of life) after, and the costs of digital subtraction angiography (DSA) with those of multi-detector row computed tomographic (CT) angiography as the initial diagnostic imaging test in patients with peripheral arterial disease (PAD). MATERIALS AND METHODS: Institutional medical ethics committee approval and patient informed consent were obtained. Between April 2000 and August 2001, patients with PAD were randomly assigned to undergo either DSA or multi-detector row CT angiography as the initial diagnostic imaging test. Outcomes were the therapeutic confidence assessed by physicians (on a scale from 0 to 10), the need for additional imaging, the health-related quality of life at 6-month follow-up, diagnostic and therapeutic costs, and the costs for a hospital stay. Costs were computed from a hospital perspective according to Dutch guidelines for cost calculations in health care. Mean outcomes were compared between groups with unpaired t testing and were adjusted for predictive baseline characteristics with multivariable regression analysis. RESULTS: Among the 145 patients, 72 were randomly allocated to the DSA group and 73 to the CT angiography group. One patient in the DSA group had to be excluded. Mean age was 63 years in the DSA group and 64 years in the CT angiography group. There were 47 men in the DSA group and 58 men in the CT angiography group. Physician confidence in making a correct therapeutic choice was significantly higher at DSA (mean confidence score, 8.2) than at CT angiography (mean score, 7.2; P < .001). During 6-month follow-up, 14% less additional imaging was performed in the DSA group than in the CT angiography group (P = .3). No significant quality-of-life differences were found between groups. The diagnostic cost associated with DSA (564 +/- 210 euro [standard deviation]) was significantly higher than that associated with CT angiography (363 +/- 273 euro), a difference of -201 euro (95% confidence interval: -281 euro, -120 euro; P < .001). Therapeutic and hospitalization costs were similar for both strategies. CONCLUSION: These results suggest that use of noninvasive multi-detector row CT angiography instead of DSA as the initial diagnostic imaging test for PAD provides sufficient information for therapeutic decision making and reduces imaging costs. PMID- 16244281 TI - Retrospective power analysis: when? PMID- 16244282 TI - Percutaneous extraluminal recanalization: usefulness of false channel balloon dilation and heparin administration before true lumen reentry. PMID- 16244283 TI - Differentiation of residual tumor from benign periablational tissues after radiofrequency ablation: the role of MR imaging contrast agents. PMID- 16244284 TI - "Hot nose sign" is not so hot. PMID- 16244286 TI - Diagnostic biomarkers for stroke: a stroke neurologist's perspective. PMID- 16244287 TI - Measuring what isn't there. PMID- 16244289 TI - Altered distribution of transferrin isoforms according to serum storage conditions. PMID- 16244288 TI - Characterization of publicly available lymphoblastoid cell lines for disease associated mutations in 11 genes. PMID- 16244290 TI - Erythrocyte riboflavin for the detection of riboflavin deficiency in pregnant Nepali women. PMID- 16244291 TI - Rapid detection of nucleophosmin (NPM1) mutations in acute myeloid leukemia by denaturing HPLC. PMID- 16244292 TI - Unusual rearrangement of the alpha-globin gene cluster containing both the alpha3.7 and alphaalphaalphaanti-4.2 crossover junctions: clinical diagnostic implications and possible mechanisms. PMID- 16244293 TI - Expression of COX-2 mRNA in peripheral blood monocytes from patients with acute myocardial infarction and its significance. PMID- 16244294 TI - Measurement of pro-C-type natriuretic peptide in plasma. PMID- 16244295 TI - False-positive results in the detection of methadone in urines of patients treated with psychotropic substances. PMID- 16244297 TI - Falsely low urinary albumin concentrations after prolonged frozen storage of urine samples. PMID- 16244296 TI - The Toll-like receptor 4 Asp299Gly and Thr399Ile polymorphisms influence the late inflammatory response in human endotoxemia. PMID- 16244298 TI - Use of fully denaturing HPLC for UGT1A1 genotyping in Gilbert syndrome. PMID- 16244299 TI - Improved sensitivity of a thyrotropin receptor antibody assay. PMID- 16244300 TI - Determination of asymmetric dimethylarginine: liquid chromatography-mass spectrometry or ELISA? PMID- 16244301 TI - Short-term urine deoxypyridinoline biological variability in the first 5 years after menopause. PMID- 16244302 TI - Investigation into the origin and tumoral mass correlation of plasma Epstein-Barr virus DNA in nasopharyngeal carcinoma. PMID- 16244303 TI - Reliable and cost-effective screening of inherited heterozygosity by Zn2+-cyclen polyacrylamide gel electrophoresis. PMID- 16244304 TI - Validation of the 99th percentile cutoff independent of assay imprecision (CV) for cardiac troponin monitoring for ruling out myocardial infarction. PMID- 16244305 TI - The polymorphism c.-3279T>G in the phenobarbital-responsive enhancer module of the bilirubin UDP-glucuronosyltransferase gene is associated with Gilbert syndrome. PMID- 16244306 TI - Tumor necrosis factor-alpha-related intraperitoneal release of CA 125 in cirrhotic patients with sterile ascites. PMID- 16244307 TI - Comparison of protocols for extracting circulating DNA and RNA from maternal plasma. PMID- 16244308 TI - Fetal rhesus D mRNA is not detectable in maternal plasma. PMID- 16244309 TI - Differences in glomerular filtration rate estimates by 2 cystatin C-based equations. PMID- 16244310 TI - Determination of amniotic fluid bilirubin absorbance in specimens contaminated by blood. PMID- 16244311 TI - CYP3A5 genotype does not influence the blood concentration of tacrolimus measured with the Abbott immunoassay. PMID- 16244312 TI - Cystatin C intrapatient variability in children with chronic kidney disease is less than serum creatinine. PMID- 16244313 TI - Autosampler programming for improved sample throughput in liquid chromatography/mass spectrometry. PMID- 16244314 TI - Gearing up for genome-wide gene-association studies. AB - One of the grand challenges of human genetics to systematically map by gene association susceptibility genes for complex diseases is underway. High throughput genotyping platforms have been developed; a comprehensive map of human genetic variation (HapMap) to guide efficient marker selection is imminent and many researchers have assembled suitable cohorts of patients. Expectations are understandably high and it is timely to review the promise and pitfalls of this strategy. PMID- 16244315 TI - Genetical genomics: combining genetics with gene expression analysis. AB - The biological mechanisms that link genetic variation and its phenotypic outcome stand as a central puzzle in biology. Geneticists have usually approached this problem by trying to identify genetic variants that underlie the trait in question. Ten years ago, microarray technology opened a second front by making it possible to compare expression levels for most active genes under a variety of genetic and environmental conditions. A typical study reveals up- or down regulation of genes or pathways associated with a phenotype (case/control) or condition (treated/untreated). In the past few years, a number of groups have started to combine gene expression studies with genetic linkage analysis, leading to a new synergy between these approaches. In this strategy, expression levels are treated as quantitative phenotypes and genetic variants that influence gene expression are sought. Several studies have shown that mRNA levels for many genes are heritable, thus amenable to genetic analysis. Quantitative trait loci mapping efforts have led to the initial characterization of genetic regulation in 'cis' probably because of variants in the gene's own regulatory regions, as well as in 'trans', i.e. by loci elsewhere in the genome. The existence of some 'master regulators' that each affects expression levels of hundreds of genes is an important finding that will surely enrich our understanding of regulatory networks. Although this novel field is still developing, understanding the genetic basis of molecular phenotypes such as gene expression is expected to shed light on the intermediate processes that connect genotype to cellular and organismal traits and represents a critical step towards true systems biology. PMID- 16244316 TI - Camels and zebrafish, viruses and cancer: a microRNA update. AB - MicroRNAs (miRNAs) form an extensive class of RNA molecules that regulate gene expression at post-transcriptional level. In recent years, much progress has been made in dissection of biogenesis and functions of miRNAs. There are at least several hundred miRNA genes in the human genome, and the emerging evidence suggests that miRNAs are broadly implicated in gene regulation. Here, we review some recent advances, and particularly we discuss how comparative genomics helps to identify novel miRNA genes, how studies in zebrafish reveal roles of miRNAs in morphogenesis, how changes in miRNA expression patterns are connected with cancer and how host-virus coevolution exploits miRNA regulatory pathways. PMID- 16244317 TI - The first molecular details of ALT in human tumor cells. AB - The activation of a telomere maintenance mechanism (TMM) is indispensable for cellular immortalization, a hallmark of human cancer. Although most human cancers use telomerase as their TMM, some use an alternative lengthening of telomeres (ALT) mechanism. The latter especially include specific subtypes of soft tissue sarcomas where ALT occurs most often in tumors with complex karyotypes, astrocytic brain tumors and osteosarcomas. The prognostic significance of ALT varies according to the type of tumor. Some ALT cells have atypical features, suggesting the possibility that there is more than one ALT mechanism. ALT cells are characterized by instability at a specific minisatellite locus (although they are stable at microsatellite loci) and by high rates of telomeric recombinational exchange. We propose a revised model whereby unequal telomeric exchange and asymmetrical chromosome segregation could result in telomere length maintenance in a cell population. In at least some ALT cells, telomere maintenance requires the integrity of the MRN (MRE11-RAD50-NBS1) recombination complex and is efficiently repressed by its sequestration. Microsatellite instability (MSI) often results in disruption of MRN, so ALT may usually be incompatible with MSI. We suggest that ALT in human tumors is a dysregulated version of an aspect of normal mammalian telomere homeostasis, which may be a vestige of the TMM used by ancient eukaryotes. Understanding the molecular basis of ALT has important implications for the diagnosis and treatment of tumors that use this TMM. PMID- 16244318 TI - Mechanisms of common fragile site instability. AB - The study of common fragile sites has its roots in the early cytogenetic investigations of the fragile X syndrome. Long considered an interesting component of chromosome structure, common fragile sites have taken on novel significance as regions of the genome that are particularly sensitive to certain forms of replication stress, which are frequently rearranged in cancer cells. In recent years, much has been learned about the genomic structure at fragile sites and the cellular checkpoint functions that monitor their stability. Recent findings suggest that common fragile sites may serve as markers of chromosome damage caused by replication stress during early stages of tumorigenesis. Thus, the study of common fragile sites can provide insight not only into the nature of fragile sites, but also into the broader consequences of replication stress on DNA damage and cancer. However, despite recent advances, many questions remain regarding the normal functional significance of these conserved regions and the basis of their fragility. PMID- 16244319 TI - Pharmacogenetics/genomics and personalized medicine. AB - Despite the marked advances in drug therapy, some patients do not respond favorably or suffer severe adverse drug effects. Pharmacogenetic studies have shown that polymorphisms of drug metabolizing enzymes, transporters and receptors contribute to variable drug response. Owing to the complexity of drug actions, a broader genomics approach aims at finding new drug targets and optimizing therapy for the individual patient. However, pharmacogenomics has made only a few inroads into clinical practice to date. This review evaluates obstacles that need to be overcome. These include the complexity of mechanisms underlying drug response, given singly or in combination, uncertainty about the genetic underpinnings of complex diseases, such as cancer, diabetes, cardiovascular and mental disorders and a lack of quantitative understanding of the scope of genetic variations, even for well-studied genes. By resolving these hurdles, pharmacogenomics will yield significant, but incremental, therapeutic advances paving the way towards personalized health care. PMID- 16244320 TI - Identification of disease genes by whole genome CGH arrays. AB - Small, submicroscopic, genomic deletions and duplications (1 kb to 10 Mb) constitute up to 15% of all mutations underlying human monogenic diseases. Novel genomic technologies such as microarray-based comparative genomic hybridization (array CGH) allow the mapping of genomic copy number alterations at this submicroscopic level, thereby directly linking disease phenotypes to gene dosage alterations. At present, the entire human genome can be scanned for deletions and duplications at over 30,000 loci simultaneously by array CGH ( approximately 100 kb resolution), thus entailing an attractive gene discovery approach for monogenic conditions, in particular those that are associated with reproductive lethality. Here, we review the present and future potential of microarray-based mapping of genes underlying monogenic diseases and discuss our own experience with the identification of the gene for CHARGE syndrome. We expect that, ultimately, genomic copy number scanning of all 250,000 exons in the human genome will enable immediate disease gene discovery in cases exhibiting single exon duplications and/or deletions. PMID- 16244321 TI - Genetic basis of Joubert syndrome and related disorders of cerebellar development. AB - Over three decades have passed since Marie Joubert described the original proband for Joubert syndrome, a rare neurological disorder featuring absence of the cerebellar vermis (i.e. midline). Efforts at deciphering the molecular basis for this disease have been complicated by the clinical and genetic heterogeneity as well as extensive phenotypic overlap with other syndromes. However, progress has been made in recent years with the mapping of three genetic loci and the identification of mutations in two genes, AHI1 and NPHP1. These genes encode proteins with some shared functional domains, but their role in brain development is unclear. Clues may come from studies of related syndromes, including Bardet Biedl syndrome and nephronophthisis, for which all of the encoded proteins localize to primary cilia. The data suggest a tantalizing connection between intraflagellar transport in cilia and brain development. PMID- 16244322 TI - Susceptibility genes for complex epilepsy. AB - Common idiopathic epilepsies are, clinically and genetically, a heterogeneous group of complex seizure disorders. Seizures arise from periodic neuronal hyperexcitability of unknown cause. The genetic component is mostly polygenic, where each susceptibility gene in any given individual is likely to represent a small component of the total heritability. Two susceptibility genes have been so far identified, where genetic variation is associated with experimentally demonstrated changes in ion channel properties, consistent with seizure susceptibility. Rare variants and a polymorphic allele of the T-type calcium channel CACNA1H and a polymorphic allele and a rare variant of the GABA(A) receptor delta subunit gene have differential functional effects. We speculate that these and other as yet undiscovered susceptibility genes for complex epilepsy could act as 'modifier' loci, affecting penetrance and expressivity of the mutations of large effect in those 'monogenic' epilepsies with simple inheritance that segregate through large families. Discovery of epilepsy associated ion channel defects in these rare families has opened the door to the discovery of the first two susceptibility genes in epilepsies with complex genetics. The susceptibility genes so far detected are not commonly involved in complex epilepsy suggesting the likelihood of considerable underlying polygenic heterogeneity. PMID- 16244323 TI - Tuberous sclerosis: a GAP at the crossroads of multiple signaling pathways. AB - Tuberous sclerosis complex (TSC) is an autosomal dominant disorder that is characterized by benign tumors (hamartomas and hamartias) involving multiple organ systems, due to inactivating mutations in TSC1 or TSC2. Here, we review recent advances in our understanding of the growth and signaling functions of the TSC1 and TSC2 proteins. Led by seminal studies in Drosophila, the TSC1/TSC2 complex has been positioned in an ancestrally conserved signaling pathway that regulates cell growth. TSC1/TSC2 receives inputs from at least three major signaling pathways in the form of kinase-mediated phosphorylation events that regulate its function as a GTPase activating protein (GAP): the PI3K-Akt pathway, the ERK1/2-RSK1 pathway and the LKB1-AMPK pathway. TSC1/TSC2 functions as a GAP towards Rheb, which is a major regulator of the mammalian target of rapamycin (mTOR). In the absence of either TSC1 or TSC2, high levels of Rheb-GTP lead to constitutive activation of mTOR-raptor signaling, thereby leading to enhanced and deregulated protein synthesis and cell growth. As a specific inhibitor of mTOR, rapamycin has therapeutic potential for the treatment of TSC hamartomas. PMID- 16244325 TI - The genetics of Fraser syndrome and the blebs mouse mutants. AB - Fraser syndrome is a recessive multisystem disorder characterized by embryonic epidermal blistering, cryptophthalmos, syndactyly, renal defects and a range of other developmental abnormalities. More than 17 years ago, the family of four mapped mouse blebs mutants was proposed as models of this disorder, given their striking phenotypic overlaps. In the last few years, these loci have been cloned, uncovering a family of three large extracellular matrix proteins and an intracellular adapter protein which are required for normal epidermal adhesion early in development. The proteins have also been shown to play a crucial role in the development and homeostasis of the kidney. We review the cloning and characterization of these genes and explore the consequences of their loss. PMID- 16244326 TI - The genetics of attention deficit hyperactivity disorder. AB - Attention deficit hyperactivity disorder (ADHD) is a highly heritable, disruptive, childhood-onset condition, the aetiology and pathogenesis of which is poorly understood. There have been relatively few genome-wide linkage studies, and no chromosomal region has yet been unequivocally implicated. In contrast, evidence from pharmacological, neuroimaging and animal studies has suggested the involvement of specific neurotransmitter systems, notably dopaminergic pathways, in ADHD and these aetiological clues have inspired a fruitful application of the candidate gene association approach. Meta-analyses or pooled data analyses have supported association between ADHD and polymorphisms in DRD4, DRD5 and SLC6A3 which encode dopamine D4 and D5 receptors and the dopamine transporter, respectively. A weaker, but nevertheless replicated, body of evidence also supports associations with SNAP-25 (synaptosomal-associated protein, 25 kDa) and SLC6A4 (serotonin transporter). There is increasing research interest in gene phenotype links, clinical phenotypic markers of heterogeneity and gene- environment interaction, which are likely to be important in the next generation of genetic studies. PMID- 16244324 TI - Insights into X-linked retinitis pigmentosa type 3, allied diseases and underlying pathomechanisms. AB - In the past decade, we have witnessed great advances in the identification of genes underlying numerous neurodegenerative diseases and the stark complexity determining genotype-phenotype relationships that lead to the impairment, and ultimately, premature death of neurons. However, significant challenges lie ahead in understanding the pathobiological and spatiotemporal processes triggered by genetic lesions underlying neurodegenerative disorders. Neuroretinal dystrophies occupy a prominent place among neurodegenerative diseases, because of the large number and prevalence of disease-causing genes, the diverse functions, the wealth of allelic, non-allelic and clinical heterogeneities determining the phenotypic expressivity and penetrance of the disease and the ease of use of animal models to probe gene function and disease pathogenesis in a well-defined neuroretinal circuitry. Retinitis pigmentosa (RP) has a prevalence of about one in 4000. RP is a retinal dystrophy leading primarily to the progressive death of photon capturing neurons--the rod photoreceptors. X-linked retinitis pigmentosa type 3 (XlRP3) accounts up to 14% of all RP cases, higher than any other single RP locus identified to date, and considered to be the most severe of all RP cases. The XlRP3 encodes the retinitis pigmentosa GTPase regulator (RPGR). RPGR interacts with the RPGR interacting protein-1 (RPGRIP1). Mutations in RPGRIP1 cause Leber's congenital amaurosis. This review highlights the progress devoted to understand the pathogenesis associated with XlRP3 and allied disorders and, concepts, trends and discrepancies emerging as molecular, subcellular and physiological processes linked to RPGR and RPGRIP1-protein network begin to be elucidated, and that may serve as a paradigm for other biological processes and neurodegenerative diseases. PMID- 16244327 TI - Emerging functions of mammalian mitochondrial fusion and fission. AB - Mitochondria provide a myriad of services to the cell, including energy production, calcium buffering and regulation of apoptosis. How these diverse functions are coordinated among the hundreds of mitochondria in a given cell is largely unknown, but is probably dependent on the dynamic nature of mitochondria. In this review, we explore the latest developments in mitochondrial dynamics in mammals. These studies indicate that mitofusins and OPA1 are essential for mitochondrial fusion, whereas Fis1 and Drp1 are essential for mitochondrial fission. The overall morphology of the mitochondrial population depends on the relative activities of these two sets of proteins. In addition to the regulation of mitochondrial shape, these molecules also play important roles in cell and tissue physiology. Perturbation of mitochondrial fusion results in defects in mitochondrial membrane potential and respiration, poor cell growth and increased susceptibility to cell death. These cellular observations may explain why mitochondrial fusion is essential for embryonic development. Two inherited neuropathies, Charcot-Marie-Tooth type 2A and autosomal dominant optic atrophy, are caused by mutations in mitofusin 2 and OPA1, suggesting that proper regulation of mitochondrial dynamics is particularly vital to neurons. Mitochondrial fission accompanies several types of apoptotic cell death and appears important for progression of the apoptotic pathway. These studies provide insight into how mitochondria communicate with one another to coordinate mitochondrial function and morphology. PMID- 16244328 TI - Small molecule intervention in microtubule-associated human disease. AB - Microtubules are essential for a number of cellular processes that include the transport of intracellular cargo or organelles across long distances and the assembly of the mitotic spindle. The identification of numerous microtubule associated proteins and the progressive elucidation of the mechanisms of microtubule assembly and transport are beginning to have a profound impact on the study and treatment of human genetic disease. A number of seemingly unrelated phenotypes have now been linked to microtubular dysfunction, especially in systems dependent heavily on microtubule-based transport, such as neurons and ciliated cells. In parallel, the association of microtubule transport defects with human genetic disease has led to the realization that targeting various aspects of microtubular biology with small molecules might offer new therapeutic paradigms, including the development of new therapeutic utility for seemingly old drugs. In this review, we discuss the use of small molecules in the investigation of microtubule-associated processes and particularly the screens of chemical compound libraries for the identification of lead compounds with potential utility in microtubule-associated disease processes. PMID- 16244329 TI - Pre hospital care in Europe. PMID- 16244330 TI - Clinicians taking pictures--a survey of current practice in emergency departments and proposed recommendations of best practice. AB - The primary objective of this survey was to establish current practice in emergency departments in the UK. Variation in obtaining consent, how image collection is achieved, and the images stored were considered to be important outcomes. An initial postal questionnaire followed by phone survey posed questions about practical and procedural issues when capturing clinical images in emergency departments in the UK. Altogether, 117 departments replied out of 150 surveyed. Only 21 departments have a written policy permitting medico-legal case photography. A total of 53 do take clinical photographs where no policy exists, seven of which actively take assault/domestic violence images, only four of which document consent. All departments with photographic facilities take images for clinical/teaching purposes. Thirty two of those without a policy attach the photograph to the clinical notes and so may be potentially called upon for medico legal proceedings if relevant, which raises issues of adequate consent procedures, storage, and confidentiality. This is particularly pertinent with the increasing use of digital photography and image manipulation. A large variation in current practice has been identified in relation to a number of issues surrounding clinical image handling in emergency departments. Subsequently, recommendations for best practice have been proposed to protect both the patient and the clinician with regards to all forms of photography in the emergency department setting. PMID- 16244331 TI - When should we thrombolyse patients with pulmonary embolism? A systematic review of the literature. AB - The early mortality in pulmonary embolism (PE) is largely predicted by the associated cardiovascular response, with progressive right ventricular failure, hypotension, shock, and circulatory arrest being associated with increasing mortality. Thrombolysis may improve the prognosis of PE associated with these varying degrees of circulatory collapse, but has no place in the treatment of small emboli with no cardiovascular compromise, as it carries a significant risk of haemorrhage. This review sets out to guide the emergency physician in deciding which patients with PE may benefit from thrombolysis. PMID- 16244332 TI - Non-invasive ventilation as a first-line treatment for acute respiratory failure: "real life" experience in the emergency department. AB - OBJECTIVE: To describe our experience with non-invasive ventilation (NIV) for patients with acute respiratory failure (ARF) in the emergency department (ED). METHODS: A prospective/retrospective, observational study on 190 patients with ARF (mean +/-SD age 72.2+/-12.9 years, mean APACHE II score 18.9+/-5.9), who received 200 NIV trials in an ED. We analysed the NIV register data (prospectively collected) and medical records (retrospective data abstraction) and evaluated clinical indications for NIV, patient outcomes, and predictive factors for success and death. NIV success was defined as tolerance of the procedure and no need for endotracheal intubation (ETI). RESULTS: Main indications to NIV were cardiogenic pulmonary oedema (CPE) (70 trials), acute exacerbation of COPD (39), both CPE and acute exacerbation of COPD (11), pneumonia (48), decompensation of obesity/hypoventilation (6), other conditions (26). The procedure was successful in 60.5% of trials. Global mortality was 34.5%, similar to the APACHE II predicted mortality of 32%. ETI rates were 6.5% and tracheostomy rates 1%. The improvement of pH within six hours after NIV initiation was predictive of survival in the hypercapnic group. CONCLUSIONS: Our results confirm the global efficacy of NIV in an ED setting, and show that, in spite of lower success rate in "real practice" in comparison with RCTs, an intermediate care unit can represent an appropriate and less expensive setting to perform this technique. The low rate of ETI seems to be because of the high number of patients for whom NIV was used as "ceiling" treatment. PMID- 16244333 TI - Relation between epistaxis, external nasal deformity, and septal deviation following nasal trauma. AB - OBJECTIVES: To find if the presence of epistaxis after nasal trauma can be used to predict post-traumatic external nasal deformity or a symptomatic deviated nasal septum. METHODS: Retrospective analysis of all patients seen in the fractured nose clinic by the first author between 17 October 2003 and 27 February 2004. Presence of epistaxis, newly developed external nasal deformity, and the presence of a deviated nasal septum with new symptoms of nasal obstruction were noted. RESULTS: A total of 139 patients were included in the study. Epistaxis following injury was noted in 106 (76%). Newly developed external nasal deformity was noted in 71 (51%), and 33 (24%) had a deviated nasal septum with new symptoms of nasal obstruction. Of the 106 patients with post-trauma epistaxis, 50 (67%) had newly developed external nasal deformity and of the 33 patients without post traumatic epistaxis, 11 (33%) had nasal deformity (p<0.05). Post-trauma epistaxis was not associated with the presence of a newly symptomatic deviated septum (25% in patients with epistaxis after injury versus 18% if there was no epistaxis). CONCLUSIONS: Presence of epistaxis after nasal trauma is associated with a statistically significant increase in external nasal deformity. However, one third of patients without epistaxis following nasal trauma also had external nasal deformity and hence all patients with a swollen nose after injury, irrespective of post-trauma epistaxis, still need to be referred to the fractured nose clinic. PMID- 16244334 TI - How is deep vein thrombosis diagnosed and managed in UK and Australian emergency departments? AB - BACKGROUND: Recent research has identified technologies that may be of value in the diagnosis and management of deep vein thrombosis (DVT). We aimed to survey current practice in the United Kingdom (UK) and Australia to determine the extent to which these technologies have been implemented in these two healthcare systems. METHODS: We undertook a postal survey of 255 hospitals in the UK and 89 hospitals in Australia, requesting details of individual diagnostic tests, use of diagnostic algorithms, and management of DVT. RESULTS: We received replies from 186/255 UK hospitals (73%) and 84/89 of Australian hospitals (94%). Ultrasonography and laboratory based D-dimer were the most commonly available tests. We received 43 different algorithms from 51 hospitals. With only a very few exceptions, DVT diagnosis was ruled in by positive venography or positive ultrasound without venographic confirmation. By contrast a variety of different criteria were used to rule out DVT. Most algorithms used a combination of low clinical risk and negative D-dimer to rule out DVT, but some required all patients to receive ultrasound or venography. Few ruled out on the basis of low clinical risk or negative D-dimer alone. Low molecular weight heparins were overwhelmingly the treatment of choice for established DVT. Most departments (214/264; 81%) offered outpatient treatment. CONCLUSION: Recently developed technologies for the diagnosis and treatment of DVT have been widely implemented in the UK and Australia. Variation in practice, and thus presumably uncertainty, seems to be greatest in relation with the criteria used to rule out DVT. PMID- 16244335 TI - The usefulness of trauma scores in determining the life threatening condition of trauma victims for writing medical-legal reports. AB - OBJECTIVES: In the Turkish legal system the severity of the victim's injury determines the severity of the criminal penalty, and the life threatening condition stated in the medical-legal report is one of the main determinants for injury severity. The aim of this study is to investigate the effectiveness and usefulness of the trauma scores in determining the life threatening condition of trauma victims from the forensic aspect in order to write accurate medical-legal reports. METHODS: Data of 296 forensic cases with blunt and penetrating trauma were obtained. The life threatening condition of patients stated in the medical legal reports according to the criteria based on traditional forensic opinion were examined. For each case, Injury Severity Score (ISS), Revised Trauma Score (RTS), and Trauma and Injury Severity Score (TRISS) were calculated. The ROC curve analysis was used to investigate the success of the trauma scores in distinguishing patients with/without life threatening conditions. Logistic regression analysis was performed to measure the association between trauma scores and life threatening conditions. RESULTS: The relations between all scores and groups (with and without life threatening risk) were found statistically significant. ISS was the most successful method in distinguishing traumatised patients both in a life threatening or non life threatening condition. CONCLUSIONS: Trauma scores can be used for making more objective, standardised, and accurate judgement on whether the injury was a life threatening one or not. These advantages of using trauma scores in such situations will also be helpful for the conclusion of the lawsuits shortly, but further studies are needed to confirm these findings. PMID- 16244336 TI - The right place in the right space? Awareness of site for needle thoracocentesis. AB - BACKGROUND: Invasive practical procedures require identification of surface anatomical landmarks to reduce risk of damage to other structures. Needle thoracocentesis has specific complications, which have been previously documented. An observational study was performed among emergency physicians to name the landmark for needle thoracocentesis and identify this point on a human volunteer as per Advanced Trauma and Life Support (ATLS) guidelines. RESULTS: A cohort of 25 emergency physicians was studied, 21 (84%) of which were ATLS certified. The correct landmark was named by 22 (88%). Only 15 (60%) correctly identified the second intercostal space on the human volunteer, all placing the needle medial to the midclavicular line, with a range of 3 cm. Two (8%) named and identified the site of needle pericardiocentesis; one (4%) named and identified the fifth intercostal space in the anterior axillary line. DISCUSSION: These results demonstrate a low accuracy among emergency physicians in identifying correct landmarks for needle thoracocentesis under elective conditions. Should greater emphasis be placed on competency based training in ATLS? PMID- 16244337 TI - Unnecessary laparotomy by using physical examination and different diagnostic modalities for penetrating abdominal stab wounds. AB - BACKGROUND: The modern management of penetrating abdominal trauma has decreased the incidence of unnecessary laparotomy by using selective non-operative management protocols. However, the real benefits of physical examination and different diagnostic methods are still unclear. METHODS: From January 2000 to April 2003, we prospectively collected data on 117 patients with penetrating stab wounds to the thoracoabdominal, anterior abdominal, and back regions who had non operative management. Clinical examination was the primary tool to differentiate those patients requiring operation. Findings of physical examination, ultrasound, computed tomography, endoscopy, echocardiography, diagnostic peritoneal lavage, and diagnostic laparoscopy were reviewed. The number of therapeutic, non therapeutic, and negative laparotomies were recorded. RESULTS: Non-operative management was successful in 79% of patients. There were 11 early (within 8 hours of admission) and 14 delayed (more than 8 hours after admission) laparotomies performed, depending on the results of various diagnostic procedures. Non operative management failed in 21% of patients, and the rate of non-therapeutic laparotomy in early and delayed laparatomy groups was 9% and 14% respectively. There was no negative laparatomy. CONCLUSIONS: The use of physical examination alone and/or together with different diagnostic methods allows reduction of non therapeutic laparotomies and elimination of negative laparatomies. PMID- 16244338 TI - Emergency department organisation of critical care transfers in the UK. AB - OBJECTIVES: Transport of the critically ill patient to or from the emergency department (ED) is a frequent occurrence. This study was designed to determine whether UK EDs currently have appropriate equipment, monitoring, staff training systems, and processes of care for transportation of the critically ill patient. METHODS: A postal questionnaire regarding ED transfer patients was sent to 247 UK EDs, followed by repeat mailing and telephone follow up of non-responders. RESULTS: In total, 139 EDs (56%) responded. An estimated 20-30 critically ill patients are transferred from and <20 are received by each ED annually. Processes of care are poorly developed; only 79 EDs (56%) have transfer guidelines available. Audit of transfers is ongoing in 59 EDs (42%), and critical incident reporting is ongoing in 122 (88%). There is a lack of immediately available transport equipment; for example, 17 EDs (12%) have no transport ventilator, 9 (6%) have no transport monitor, and 9 (6%) have no syringe pump. Transport equipment is invariably not standardised. Anaesthetic staff of specialist registrar (74 doctors; 53%) or senior house officer (36 doctors; 26%) grades carry out the majority of ED transfers accompanied by a D or E grade nurse. Both invariably have no formal transfer training. CONCLUSIONS: This study highlights inadequacies in provision of equipment and monitoring during interhospital transfer from the ED. Training and processes of care for transport of the critically ill are also suboptimum. Many departments are currently reviewing these processes to formalize and improve transfer training procedures and protocols. PMID- 16244339 TI - Self-harm, capacity, and refusal of treatment: implications for emergency medical practice. A prospective observational study. AB - OBJECTIVES: In the context of increasing attention to the rights of adults to make treatment decisions for themselves, this study investigated, among patients who have engaged in self-harm (i) the extent of valid decision making; (ii) the impact of mental disorders; and (iii) the effect of systematically providing relevant clinical information. DESIGN: A prospective observational study. SETTING: The emergency department of a large teaching hospital in southeast England. PARTICIPANTS: Seventy one adult men and women who had presented for treatment following self-harm. MAIN OUTCOME MEASURES: Semi-structured interviews were used to make clinical judgements about participants' capacity to consent before, and following, the presentation of simple written information about the proposed treatment(s). Demographic data, and data about mental disorder and alcohol misuse, were also collected. RESULTS: Based on accepted legal criteria, only 28/71 (39.4%) of the patients were judged to have capacity to consent to the proposed intervention(s) initially. However, the number of patients judged to have capacity improved significantly (p<0.001) after the presentation of written information, to 45/71 (63.4%). Those judged incapable were significantly more likely (p<0.01) to refuse treatment. Continuing incapacity was significantly associated only with cognitive impairment (p<0.001) and/or severe psychiatric disturbance (p<0.01). CONCLUSIONS: Consistent with current views, capacity is not static, even among patients who have engaged in self-harm, but can be improved through a simple intervention. The findings are consistent with recent guidance about supporting this vulnerable group of patients, many of whom are ambivalent about treatment. PMID- 16244340 TI - "Honey moon" meningitis. AB - Streptococcus agalactiae is a rare cause of meningitis in healthy non-pregnant adults. A case of S. agalactiae meningitis is reported in a previously healthy young woman following sexual intercourse for the first time. The vaginal flora was the verified source of infection. PMID- 16244341 TI - An audit of symptom triggered chlordiazepoxide treatment of alcohol withdrawal on a medical admissions unit. AB - Patients suffering from alcohol withdrawal were found on initial audit to be discharged before the resolution of their symptoms. A regimen using symptom triggered chlordiazepoxide was introduced and found, on re-audit and anecdotally, to be an improvement. The duration of symptoms was reduced significantly, the length of stay was reduced (although not statistically significant because of limited power), and most patients who had experience of both regimens found the symptom triggered regimen more effective. PMID- 16244342 TI - Reduction of dislocated hip prosthesis in the emergency department using conscious sedation: a prospective study. AB - This prospective observational study aimed to determine the rate of success of reduction of dislocated hip prostheses using conscious sedation. In 101 consecutive patients presenting to the emergency department between August 2000 and February 2003 with a dislocated hip prosthesis, reduction was attempted using conscious sedation. The outcome measures of the study were (a) rate of success of the attempted reductions (b) rate of complication of the sedation or the procedure, and (c) rate of success in the three subgroups (based on degree of dislocation). The overall success rate was 62% (95% CI 53% to 71%). There were six complications: five related to sedation and one was a mild foot drop. The mean time to attempted reduction using conscious sedation was 1.83 hours and for an equivalent group who were excluded and subsequently required general anaesthesia the mean time was 10.9 hours. Reduction of isolated unilateral prosthetic hip dislocation using conscious sedation in the emergency department is safe and has a reasonable success rate. Prosthetic hip reduction can be attempted more quickly using conscious sedation than when awaiting general anaesthesia. PMID- 16244343 TI - Simple measures to reduce the rate of contamination of blood cultures in Accident and Emergency. AB - OBJECTIVES: To reduce the contamination rate of blood cultures taken in the Accident and Emergency (A&E) department. METHODS: The standard blood culture sampling kit was supplemented with an instruction sheet on the optimal method for drawing blood cultures and a large 62% ethyl alcohol impregnated wipe. RESULTS: There was a statistically significant reduction in the number of contaminants (p = 0.03). CONCLUSIONS: Simple measures to encourage skin disinfection and appropriate sampling technique will reduce the incidence of contamination of blood cultures in the A&E department. PMID- 16244346 TI - Best evidence topic report. How to immobilise after shoulder dislocation? AB - A short cut review was carried out to establish the best way to immobilise dislocated shoulders after reduction. A total of 47 papers were identified using the reported search, of which four represent the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results, and study weaknesses of these best papers are tabulated. For patients with a first anterior shoulder dislocation immobilisation in external rotation may be of more benefit than immobilisation in internal rotation. PMID- 16244344 TI - Best evidence topic report. Incision and drainage preferable to oral antibiotics in acute paronychial nail infection? AB - A short cut review was carried out to establish whether incision and drainage or antibiotics was best for acute paronychia. No relevant papers were found using the reported search. There is currently no evidence that oral antibiotics are any better or worse than incision and drainage for acute paronychiae. PMID- 16244347 TI - Best evidence topic report. No evidence found that a femoral nerve block in cases of femoral shaft fractures can delay the diagnosis of compartment syndrome of the thigh. AB - A short cut review was carried out to establish whether a femoral nerve block may mask the signs and symptoms of thigh compartment syndrome. No relevant papers were found using the reported search. There is no evidence to associate a femoral nerve block with a delayed or missed diagnosis of compartment syndrome. PMID- 16244348 TI - Best evidence topic report. Cricoid pressure in emergency rapid sequence induction. AB - A short cut review was carried out to establish cricoid pressure reduced aspiration during rapid sequence induction (RSI) of anaesthesia. A total of 241 papers were identified using the reported search, of which three represented the best evidence to answer the clinical question. The author, date and country of publication, patient group studied, study type, relevant outcomes, results, and study weaknesses of these best papers are tabulated. There is little evidence to support the widely held belief that the application of cricoid pressure reduces the incidence of aspiration during a rapid sequence intubation. PMID- 16244350 TI - Code of practice between immediate care doctors and ambulance NHS trusts. AB - All medical practitioners working within the NHS are subject to annual appraisal and in the future revalidation. Medical responders acting on behalf of NHS Ambulances Trusts often working in a voluntary capacity will require appraisal either individually (those working exclusively in prehospital care) or jointly (for hospital practitioners or GP's who are also involved prehospital care). They will be required to demonstrate satisfactorily adherence to the principles of clinical governance and develop a CPD portfolio. Currently there exists no specific code of practice for medical practitioners responding on behalf of the ambulance service. To establish a mutual understanding between the Ambulance Services and medical practitioners the Faculty of Prehospital Care has met with stakeholders to establish a code of practice which has been agreed and endorsed by the Ambulance Service Association. This document clearly defines both individual and joint responsibilities to establish good practice. For many practitioners much of what is contained in this document will already be in place. PMID- 16244349 TI - Extrication of the seriously injured road crash victim. AB - In any accident scene, rapid removal of the casualty to hospital improves their chances of survival. The term the "Golden Hour" was first introduced in 1961, but because of misinterpretation as to what period this actually referred to, a second concept, the "Platinum Ten Minutes" was proposed as the time taken to move a casualty to the ambulance. To achieve this rapid removal, the ambulance and medical personnel must work in harmony with the police and fire service to secure the scene and remove the casualty safely without causing injury either to the casualty or other personnel on the scene. Care should also be taken to preserve evidence at the scene for future investigation. This paper describes the role of the emergency services, in particular that of the fire officer in charge, when attending the scene of a road traffic accident. The methods and equipment used to extricate a casualty from a motor vehicle are explained. PMID- 16244351 TI - Atrial fibrillation after vardenafil therapy. AB - Vardenafil is a new oral phosphodiesterase inhibitor used for erectile dysfunction. We report a case admitted with a first-detected, symptomatic paroxysmal atrial fibrillation in a healthy patient after self-medication with vardenafil. PMID- 16244352 TI - "My back is killing me". AB - This is a case of a rare cause of back pain that presented to several clinicians before the diagnosis became obvious and the correct management was initiated. Lemierre's syndrome was confirmed at post mortem examination. We conclude that thoracic back pain should not be assumed to have a simple mechanical cause and that efforts should be made to discover the underlying aetiology. PMID- 16244353 TI - Pneumothorax following central venous catheterisation? AB - A chest radiograph performed to check the position of a central venous catheter in a patient appeared to show a pneumothorax. Intercostal drain insertion was prepared. Reassessment of the patient and a further radiograph confirmed that the "pneumothorax" was an artefact from a prominent skin fold due to the patient's body habitus. PMID- 16244354 TI - Bilateral optic disc swelling; is a CT scan necessary? AB - A 47 year old man sustained a head injury after tripping. He presented to the accident and emergency department next morning where head x ray revealed no fractures. However, the casualty doctor found bilateral blurred optic disc margins on ophthalmoscopy. Although his head injury was classed as non-serious, an urgent computed tomography scan was ordered and an ophthalmic opinion sought. After detailed retinal examination, the ophthalmologist made a diagnosis of bilateral optic nerve head drusen (ONHD), which was confirmed by a B-scan ultrasound. The patient was advised not to drive (due to constricted visual fields associated with ONHD) and to inform his siblings of his condition so they could also be assessed. In cases of apparent optic disc swellings, it is essential to distinguish between true and pseudo-papillo-oedema to avoid subjecting patients to unnecessary neuroimaging procedures and associated exposure to radiation. PMID- 16244355 TI - Purtscher's retinopathy after fracture dislocation of shoulder joint. AB - Purtscher described sudden blindness in patients with severe head trauma due to a remote retinopathy, characterised by bilateral retinal haemorrhages, cotton wool spots, and optic disc swelling seen on fundoscopy. A similar retinopathy has been reported in compressive chest trauma, long bone fractures, and acute pancreatitis. It is less well recognised that Purtscher's retinopathy can occur unilaterally and following less severe trauma. We present a case of unilateral remote traumatic retinal angiopathy following a fracture dislocation of the shoulder joint. PMID- 16244357 TI - Eosinophilic gastroenteritis presenting as relapsing severe abdominal pain and enteropathy with protein loss. AB - Eosinophilic gastroenteritis (EG) is an uncommon gastrointestinal tract disease, and diagnosis can be difficult. A combination of acute or recurrent abdominal pain with peripheral hypereosinophilia suggests the diagnosis. Surgery in patients with these features might therefore be avoided. Physicians must maintain a high index of suspicion and a working knowledge of the natural history of EG in order to establish the proper diagnosis. We present the case of a young man with EG who presented with relapsing severe abdominal pain and enteropathy with protein loss. PMID- 16244356 TI - Modified release verapamil induced cardiogenic shock. AB - Cardiogenic shock due to acute myocardial infarction is commonly seen in the accident and emergency department. Refractory cardiogenic shock has been reported after a therapeutic dose of modified release verapamil with concomitant use of beta blocker, metoprolol, but not after a single therapeutic oral dose of modified release verapamil alone. We report what we believe to be the first case of potentially life threatening cardiogenic shock resulting from the myocardial depressant effect of a single therapeutic oral dose of modified release verapamil. The patient made a dramatic recovery minutes after an injection of intravenous calcium chloride. The case is a reminder of the negative inotropic effect of verapamil and how it should be treated. PMID- 16244359 TI - The joint effect of smoking and AIB1 on breast cancer risk in BRCA1 mutation carriers. AB - Women with BRCA1 mutations are at an elevated risk for breast cancer. AIB1 (NCOA3/SRC3) genotype and smoking may alter this risk. We examined the differences in breast cancer risk by AIB1 polyglutamine repeat polymorphism and pre-diagnosis smoking habits for BRCA1 mutation carriers to determine if there was an interaction between smoking and AIB1 genotype. Multivariate Cox proportional hazards regression was used with 316 female BRCA1 mutation carriers to model breast cancer risk. Ever having smoked was associated with a decreased breast cancer risk [Hazard Ratio (HR) = 0.63, 95% CI, 0.47-0.87]. A dose-response relationship between number of pack-years smoked and breast cancer risk was also found for women who smoked <20 pack years of cigarettes (HR = 0.72, 95% CI, 0.52 1.00) and for women who smoked >/=20 pack years (HR = 0.41, 95% CI, 0.23-0.71; P for trend = 0.0007). Women with a 28 repeat allele for AIB1 had a significantly reduced risk of breast cancer (HR = 0.72, 95% CI, 0.51-1.00). Women who smoked >/=20 pack-years with a 28 repeat allele had an even greater reduced risk of breast cancer (HR = 0.19, 95% CI, 0.07-0.54) compared to women who were never smokers with no 28 allele. Since AIB1 appears to modulate the effect of endogenous hormones via the estrogen receptor, and smoking affects circulating hormone levels, these results support evidence that steroid hormones play an important role in breast carcinogenesis in BRCA1 mutation carriers, and suggest mechanisms for developing novel cancer prevention strategies for BRCA1 mutation carriers. PMID- 16244360 TI - Role of the main access channel of catalase-peroxidase in catalysis. AB - Catalase-peroxidases (KatG) are bifunctional heme peroxidases with an overwhelming catalatic activity. The structures show that the buried heme b is connected to the exterior of the enzyme by a main channel built up by KatG specific loops named large loop LL1 and LL2, the former containing the highly conserved sequence Met-Gly-Leu-Ile-Tyr-Val-Asn-Pro-Glu-Gly. LL1 residues Ile248, Asn251, Pro252, and Glu253 of KatG from Synechocystis are the focus of this study because of their exposure to the solute matrix of the access channel. In particular, the I248F, N251L, P252A, E253Q, and E253D mutants have been analyzed by UV-visible and resonance Raman spectroscopies in combination with steady-state and presteady-state kinetic analyses. Exchange of these residues did not alter the kinetics of cyanide binding or the overall peroxidase activity. Moreover, the kinetics of compound I formation and reduction by one-electron donors was similar in the variants and the wild-type enzyme. However, the turnover numbers of the catalase activity of I248F, N251L, E253Q, and E253D were only 12.3, 32.6, 25, and 42% of the wild-type activity, respectively. These findings demonstrate that the oxidation reaction of hydrogen peroxide (not its reduction) was affected by these mutations. The altered kinetics allowed us to monitor the spectral features of the dominating redox intermediate of E253Q in the catalase cycle. Resonance Raman data and structural analysis demonstrated the existence of a very rigid and ordered structure built up by the interactions of these residues with distal side and also (via LL1) proximal side amino acids, with the heme itself, and with the solute matrix in the channel. The role of Glu253 and the other investigated channel residues in maintaining an ordered matrix of oriented water dipoles, which guides hydrogen peroxide to its site of oxidation, is discussed. PMID- 16244358 TI - Attenuation of BPDE-induced p53 accumulation by TPA is associated with a decrease in stability and phosphorylation of p53 and downregulation of NFkappaB activation: role of p38 MAP kinase. AB - DNA damage caused by benzo[a]pyrene (B[a]P) or other polynuclear hydrocarbons (PAHs) induce p53 protein as a protective measure to eliminate the possibility of mutagenic fixation of the DNA damage. 12-O-tetradecanoylphorbol-13-acetate (TPA) inhibits p53 response induced by B[a]P and other DNA-damaging agents and may cause tumor promotion. The molecular mechanism of attenuation of B[a]P-induced p53 response by TPA is not known. We investigated the effect of TPA on p53 response in (+/-)-anti-benzo[a]pyrene-7,8-diol-9,10-epoxide (BPDE)-treated mouse epidermal JB6(P(+)) Cl 41 cells. BPDE treatment induced p53 accumulation which was attenuated significantly by TPA. Cells treated with BPDE and TPA showed increased ratio of Mdm2 to p53 proteins in p53 immunoprecipitate and decreased p53 life span compared to BPDE-treated cells indicating p53 destabilization by TPA. TPA also inhibited BPDE-induced p53 phosphorylation at serine15. Activation of both ERKs and p38 MAPK by BPDE and attenuation of BPDE-induced p53 accumulation by U0126 or SB202190, specific inhibitor of MEK1/2 or p38 MAPK, indicate the role of ERKs and p38 MAPK in p53 accumulation. Interestingly, TPA potentiated BPDE-induced activation of ERKs whereas p38 MAPK activation was significantly inhibited by TPA, suggesting that inhibition of p38 MAPK is involved in p53 attenuation by TPA. Furthermore, SB202190 treatment caused decreased p53 stability and inhibition of phosphorylation of p53 at serine15 in BPDE-treated cells. We also observed that TPA or SB202190 attenuated BPDE-induced nuclear factor kappa B (NFkappaB) activation in JB6 Cl 41 cells harboring NFkappaB reporter plasmid. To our knowledge this is the first report that TPA inhibits chemical carcinogen-induced NFkappaB activation. Interference of TPA with BPDE-induced NFkappaB activation implicates abrogation of p53 function which has been discussed. Overall, our data suggest that abrogation of BPDE-induced p53 response and of NFkappaB activation by TPA is mediated by impairment of the signaling pathway involving p38 MAPK. PMID- 16244362 TI - The nucleotide-binding site of the Escherichia coli DnaC protein: molecular topography of DnaC protein-nucleotide cofactor complexes. AB - The structure of the nucleotide-binding site of the Escherichia coli replication factor DnaC protein and the effect of the nucleotide cofactor on the protein structure have been examined using ultraviolet, steady-state, and time-dependent fluorescence spectroscopy. Emission spectra and quenching studies of the fluorescent nucleotide analogs, 3'-O-(N-methylantraniloyl)-5'-triphosphate (MANT ATP) and 3'-O-(N-methylantraniloyl)-5'-diphosphate (MANT-ADP), bound to the DnaC protein indicate that the nucleotide-binding site forms a hydrophobic cleft on the surface of the protein. Fluorescence decays of free and bound MANT-ATP and MANT-ADP indicate that cofactors exist in two different conformations both, free and bound to the protein. However, the two conformations of the bound nucleotides differ in their solvent accessibilities. Moreover, there are significant differences in the solvent accessibility between ATP and ADP complexes. Specific binding of magnesium to the protein controls the structure of the binding site, particularly, in the case of the ATP complex, leading to additional opening of the binding site cleft. Both tyrosine and tryptophan residues are located on the surface of the protein. The tryptophans are clustered at a large distance from the nucleotide-binding site. However, in spite of a large spatial separation, binding of both cofactors induces significant and different changes in the structure of the environment of tryptophans, indicating long-range structural effects through the DnaC molecule. Moreover, only ATP induces changes in the distribution of the tyrosine residues on the surface of the protein. The data reveal that the nucleotide-DnaC protein complex is a sophisticated allosteric system, responding differently to the ATP and ADP binding. PMID- 16244361 TI - Identification of the major site of O-linked beta-N-acetylglucosamine modification in the C terminus of insulin receptor substrate-1. AB - Signal transduction from the insulin receptor to downstream effectors is attenuated by phosphorylation at a number of Ser/Thr residues of insulin receptor substrate-1 (IRS-1) resulting in resistance to insulin action, the hallmark of type II diabetes. Ser/Thr residues can also be reversibly glycosylated by O linked beta-N-acetylglucosamine (O-GlcNAc) monosaccharide, a dynamic posttranslational modification that offers an alternative means of protein regulation to phosphorylation. To identify sites of O-GlcNAc modification in IRS 1, recombinant rat IRS-1 isolated from HEK293 cells was analyzed by two complementary mass spectrometric methods. Using data-dependent neutral loss MS3 mass spectrometry, MS/MS data were scanned for peptides that exhibited a neutral loss corresponding to the mass of N-acetylglucosamine upon dissociation in an ion trap. This methodology provided sequence coverage of 84% of the protein, permitted identification of a novel site of phosphorylation at Thr-1045, and facilitated the detection of an O-GlcNAc-modified peptide of IRS-1 at residues 1027-1073. The level of O-GlcNAc modification of this peptide increased when cells were grown under conditions of high glucose with or without chronic insulin stimulation or in the presence of an inhibitor of the O-GlcNAcase enzyme. To map the exact site of O-GlcNAc modification, IRS-1 peptides were chemically derivatized with dithiothreitol following beta-elimination and Michael addition prior to LC-MS/MS. This approach revealed Ser-1036 as the site of O-GlcNAc modification. Site-directed mutagenesis and Western blotting with an anti-O GlcNAc antibody suggested that Ser-1036 is the major site of O-GlcNAc modification of IRS-1. Identification of this site will facilitate exploring the biological significance of the O-GlcNAc modification. PMID- 16244364 TI - Conservation and variation of gene regulation in embryonic stem cells assessed by comparative genomics. AB - We have examined the gene structure and regulatory regions of octamer-binding transcription factor 3/4 (Oct 3/4), sex determining region Y box 2 (Sox2), signal transducer and activator of transcription 3 (Stat3), embryonal stem cell-specific gene 1 (ESG), Nanog homeobox (Nanog), and several other genes highly expressed in embryonic stem (ES) cells across different species. Our analysis showed that ES cell-expressed Ras (ERAS) was orthologous to a human pseudogene Harvey Ras (HRASP) and that the promoter and other regulatory sequences were highly divergent. No ortholog of (ES) cell-derived homeobox containing gene (Ehox) could be identified in human, and the closest paralogs PEPP gene subfamily 1 (PEPP1), PEPP2, and extraembryonic, spermatogenesis, homeobox 1 (Esx1) were not expressed by ES cells and shared little homology. The Sox2 promoter was the most conserved across species and the Oct3/4 promoter region showed significant homology particularly in the distal enhancer active in ES cells. Analysis suggested common and divergent pathways of regulation. Conserved Oct3/4 and Sox2 co-binding domains were identified in most ES expressed genes, highlighting the importance of this transcriptional pathway. Conserved fibroblast growth factor response element sites were identified in regulatory regions, suggesting a potential parallel pathway for regulation by FGFs. A central role of Stat3 activation in self-renewal and in a regulatory feedback loop was suggested by the identification of the conserved binding sites in most pathways. Although most pathways were evolutionarily conserved, promoters and genomic structure of the leukemia inhibitory factor (LIF) pathway components were divergent, likely explaining the differential requirement of LIF for human and rodent cells. Our analysis further suggested that the Nanog regulatory pathway was relatively independent of the LIF/Oct pathway and may interact with the Nodal/transforming growth factor-beta pathway. These results provide a framework for examining the current reported differences between rodent and human ES cells and define targets for future perturbation studies. PMID- 16244363 TI - Long QT syndrome-associated I593R mutation in HERG potassium channel activates ER stress pathways. AB - Hereditary long QT syndrome is a fatal arrhythmia associated with gene mutations in potassium and sodium channels that are expressed in ventricle. By employing heterologous expression and making comparisons to cells expressing wild-type human-ether-a-go-go-related protein (HERG), a potassium channel that contributes to I(Kr) current in ventricular cardiomyocytes, we demonstrate activation of an elevated endoplasmic reticulum (ER) stress response by the mutant I593R HERG potassium channel implicated in long QT syndrome type 2. I593R HERG is trafficking-impaired and forms Triton-insoluble aggregates. Expression of I593R HERG activates the unfolded protein response pathway and, separately, NF-kappaB signaling. ATF6, the activating transcription factor of the unfolded protein response pathway, is processed into the active transcription factor in the cells expressing I593R HERG. Consistent with ATF6 activation, the ER chaperones/calcium binding proteins Grp78, Grp94, and calreticulin are elevated in I593R HERG expressing cells. Coexpression of I593R HERG with wild-type HERG also results in ER stress pathway activation. By eliciting downstream links in signal transduction pathways associated with ER stress, expression of mutant trafficking impaired ion channels may contribute to disease etiology mechanisms that augment those associated with attenuated ion flux. PMID- 16244365 TI - Protein-surface interactions: an energy-based mathematical model. AB - This article describes an energy-based approach to protein adsorption, focusing on the energies involved in the interactions between a protein and a surface. Mathematical modeling and simulation based on this approach allow an improved understanding of the conditions that favor or prevent adsorption of a protein onto a surface and that can play a significant role in the design of material surfaces that interact with biological tissues according to specific needs. Biocompatibility with respect to fluids in motion, such as blood, is the main foreseeable application of our work. The considered energies are the van der Waals energy, the electrostatic energy, and the hydrophobic or hydrophilic energy. Moreover, the motion of the medium in which particles are immersed is also taken into account, considering the drag effect of the motion of the fluid on the particle, leading to a kinetic contribution to the total energy. It is shown that the adsorption behavior is not mainly determined by the van der Waals energy and by the double layer energy, but that a significant role is also played by the hydrophobic or hydrophilic energy. These results support the findings of experimental studies. PMID- 16244367 TI - Light-induced protein-matrix uncoupling and protein relaxation in dry samples of trehalose-coated MbCO at room temperature. AB - In humid samples of trehalose-coated carboxy-myoglobin (MbCO), thermally driven conformational relaxation takes place after photodissociation of the carbon monoxide (CO) molecule at room temperature. In such samples, because of the extreme viscosity of the external matrix, photodissociated CO cannot diffuse out of the protein and explores the whole (proximal and distal side) heme pocket, experiencing averaged protein heme pocket structures, as a result of the presence of Brownian motions. At variance, in very dry samples, a lower portion of the photodissociated CO diffuses from the distal to the proximal heme pocket side probing in nonaveraged structures. We revisit here the flash photolysis data by Librizzi et al. (2002) and report on new, room temperature experiments in MbCO trehalose samples, shortly illuminated prior the laser pulse. In dry samples, pre illumination increased the diffusion of CO from the distal to the proximal heme pocket side, which resulted in less structure than in non-pre-illuminated samples. Such an effect, which is absent in humid samples, stems from a decoupling of the protein internal degrees of freedom from those of the external water-sugar matrix. We suggest that such a decoupling can be brought about by the continuous attempts performed by the protein during pre-illumination to undergo relaxation toward the photodissociated deoxy state. This, in turn, causes a collapse in the hydrogen bond network, which connects the protein surface to the water-sugar matrix, as reported by Cottone et al. (2002) and Giuffrida et al. (2003). In the conclusion section, we discuss the possible involvement of the processes invoked to rationalize the present data, in the function of macromolecules and interactions in living cells. PMID- 16244366 TI - Gramicidin D and dithiothreitol effects on erythrocyte exovesiculation. AB - The use of either diphenylhexatriene, trimethylamino-diphenylhexatriene, or heptadecyl-hydroxycoumarin (C17-HC) allows, simultaneously and with the same molecule, the induction of erythrocyte exovesiculation and labeling of the released vesicles with the fluorescent probe. This method was used to evaluate gramicidin D (a channel-forming peptide) and dithiothreitol (a reducing agent) effects on the human erythrocytes vesiculation process. The release of cholesterol and phospholipids in exovesicles at longer incubation times was only detectable in the presence of gramicidin or dithiothreitol. When C17-HC was used to induce the vesiculation, the presence of gramicidin or dithiothreitol lead to a drastic decrease on the [phospholipids]/[cholesterol] ratio. However, in the samples with dithiothreitol, this variation did not result in the expectable decrease of membrane fluidity. These effects can be related with the presence of lipid rafts, the transbilayer lipids reorientation induced by gramicidin or dithiothreitol, and the cholesterol-dependent gramicidin channels inactivation. PMID- 16244369 TI - Mitogen-activated protein kinases in cell-cycle control. AB - The mitogen-activated protein kinase (MAPK) family of kinases connects extracellular stimuli with diverse cellular responses ranging from activation or suppression of gene expression to the regulation of cell mortality, growth, and differentiation. The MAPK family has been studied extensively; however, the role of these kinases in cell growth and cell-cycle control has become increasingly complex. Patterns have begun to emerge from these studies that show the functions of MAPK subfamilies at different stages of the cell cycle. Their patterns of subcellular localization and movement during the cell cycle are subfamily specific and have raised many questions about possible cell-cycle functions that have yet to be demonstrated. This article will compare and contrast our current understanding of the functions and localization patterns of the MAPK subfamilies (ERK, BMK, p38, and JNK) in cell-cycle control. PMID- 16244368 TI - Cytoskeletal regulation of nitric oxide synthase. AB - The three isoforms of nitric oxide synthase (NOS)--endothelial NOS (eNOS), inducible NOS (iNOS), and neural NOS (nNOS)--colocalize with the cytoskeleton including actin microfilaments, microtubules, and intermediate filaments directly or indirectly. These colocalizations enable optimal nitric oxide production and help NOS exert their functions. The reorganization of cytoskeletal polymerization state induced by extracellular stimuli such as shear stress, hypoxia, and drugs regulates eNOS, nNOS, and iNOS. Alterations of nitric oxide production caused by cytoskeletal reorganization play an important role in physiological and pathophysiological conditions. This review focuses on recent data regarding the regulation of NOS by the cytoskeleton at transcriptional, posttranscriptional, and posttranslational levels. PMID- 16244370 TI - Atherosclerosis and cancer: flip sides of the neoplastic response in mammalian cells? AB - Scientific and medical evidence over the past 30 years has established striking parallels between atherosclerosis and cancer--pathogenetic relationships that cross the boundaries of fiction into the realm of reason. Both diseases in humans are characterized by uncontrolled regulation of cellular growth and differentiation and share many common genomic targets during the course of growth dysregulation. Such parallels can be reconciled if atherosclerotic plaques are viewed as neoplasms of smooth muscle origin. PMID- 16244371 TI - Dietary vitamin E decreases doxorubicin-induced oxidative stress without preventing mitochondrial dysfunction. AB - Doxorubicin (DOX) is a widely prescribed antineoplastic and although the precise mechanism(s) have yet to be identified, DOX-induced oxidative stress to mitochondrial membranes is implicated in the pathogenic process. Previous attempts to protect against DOX-induced cardiotoxicity with alpha-tocopherol (vitamin E) have met with limited success, possibly as a result of inadequate delivery to relevant subcellular targets such as mitochondrial membranes. The present investigation was designed to assess whether enrichment of cardiac membranes with alpha-ocopherol is sufficient to protect against DOX-induced mitochondrial cardiotoxicity. Adult male Sprague-Dawley rats received seven weekly subcutaneous injections of 2 mg/kg DOX and fed either standard diet or diet supplemented with alpha-tocopherol succinate. Treatment with a cumulative dose of 14 mg/kg DOX caused mitochondrial cardiomyopathy as evidenced by histology, accumulation of oxidized cardiac proteins, and a significant decrease in mitochondrial calcium loading capacity. Maintaining rats on the alpha tocopherol supplemented diet resulted in a significant (two- to four-fold) enrichment of cardiac mitochondrial membranes with alpha-tocopherol and diminished the content of oxidized cardiac proteins associated with DOX treatment. However, dietary alpha-tocopherol succinate failed to protect against mitochondrial dysfunction and cardiac histopathology. From this we conclude that although dietary vitamin E supplementation enriches cardiac mitochondrial membranes with alpha-tocopherol, either (1) this tocopherol enrichment is not sufficient to protect cardiac mitochondrial membranes from DOX toxicity or (2) oxidative stress alone is not responsible for the persistent mitochondrial cardiomyopathy caused by long-term DOX therapy. PMID- 16244372 TI - Doxorubicin cardiac dysfunction: effects on calcium regulatory proteins, sarcoplasmic reticulum, and triiodothyronine. AB - Utilizing a model of chronic doxorubicin cardiomyopathy, this study examines the relationship between changes in expression and function of calcium handling proteins and contractile dysfunction. A possible mechanism to account for this relationship is suggested. New Zealand white rabbits were injected with either doxorubicin (1 mg/kg, twice weekly for 8 wk) or 0.9% NaCl. Gene transcript, protein levels, and the function of several proteins from the left ventricle were assessed. Protein levels of sarcoplasmic reticulum (SR) Ca2+ transporting ATPase (SERCA2a and b), Ca2+ release channel (RYR2), calsequestrin, Na/Ca exchanger, mRNA levels of RYR2, and [3H]-ryanodine binding (B(max)) to RYR2 were significantly decreased in doxorubicin-treated rabbits; protein levels of phospholamban, dihydropyridine receptor alpha2 subunit, and SR Ca2+ loading rates were not decreased. However, only protein levels of SERCA2 and RYR2, mRNA levels of RYR2, and Bmax of RYR2 significantly regressed with left-ventricular fractional shortening. Analysis of contractile function of atrial preparations isolated from doxorubicin-treated rabbits revealed that doxorubicin diminished contractility (dF/dt) of rest-potentiated contractions consistent with SR dysfunction. Serum concentrations of free triiodothyronine (T3) decreased in doxorubicin-treated rabbits. Our results suggest that chronic doxorubicin administration in the rabbit causes a SR-dependent contractile dysfunction that may result, in part, from decreased T3. PMID- 16244373 TI - Cardiomyocyte dysfunction in models of type 1 and type 2 diabetes. AB - Cardiomyopathy is a major cause of mortality for both type 1 and 2 diabetic patients. However, experimental analysis of diabetic cardiomyopathy has focused on type 1 diabetes and there are few reports on cardiomyocyte dysfunction in the widely used type 2 diabetic model, db/db. In the current study, we assessed function in isolated ventricular myocytes from type 1 diabetic OVE26 mice and from type 2 diabetic db/db mice. When compared with their respective control strains, both diabetic models showed significant impairment in contractility, as assessed by percent peak shortening, maximal rate of contraction, and maximal rate of relaxation. The calcium decay rate was also significantly reduced in both types of diabetes, but the decrement was much greater in OVE26 myocytes, approx 50% vs only 20% in db/db myocytes. To understand the basis for slow calcium decay in diabetic myocytes and to understand the molecular basis for the quantitative difference between calcium decay in OVE26 and db/db myocytes, we measured cardiac content of the SERCA2a calcium pump. SERCA2a was significantly decreased in OVE26 diabetic myocytes but not reduced at all in db/db myocytes. The reduction of SERCA2a in OVE26 myocytes was completely prevented by overexpression of the antioxidant protein metallothionein, confirming that oxidative stress is an important component of diabetic cardiomyopathy. The current results demonstrate that though contractility is impaired in individual myocytes of db/db hearts and deficits are similar to what is seen in a severe model of type 1 diabetes, impairment in calcium reuptake is less severe, probably as a result of maintenance of normal levels of SERCA2a. PMID- 16244374 TI - Overexpression of hexokinase protects hypoxic and diabetic cardiomyocytes by increasing ATP generation. AB - Cardiac glucose metabolism is critical to hypoxic cardiac function and hypoxia is known to stimulate glucose metabolism. This increases generation of ATP when mitochondrial respiration is inhibited. In diabetes, cardiac glucose metabolism declines and this may contribute to diabetic cardiomyopathy. The first step in committing glucose to metabolism is glucose phosphorylation catalyzed by hexokinase. But the potential role of hexokinase in the hypoxic or diabetic heart is uncertain. This study is designed to assess the ability of hexo-kinase and elevated ATP to protect cardiomyocyte contractility from hypoxia and diabetes. We used cardiomyocytes from the transgenic mouse Mh, which has cardiac specific expression of yeast hexokinase, to investigate the importance of glucose phosphorylation in the myocyte response to hypoxia and diabetes. Cardiomyocytes were isolated from FVB control and Mh hearts to assess the effects of 2 h of hypoxia on myocyte contractility and ATP content. The protective effect of hexokinase on diabetes was assessed in myocytes from the OVE26 Type I diabetic mouse and in OVE26Mh diabetic mice that carry the hexokinase gene. Overexpression of hexokinase had no effect during aerobic culture, but during hypoxia, hexokinase improved ATP content by 44% and this restored contractility almost to normal levels. In myocytes from diabetic mice, tested under both aerobic and hypoxic conditions, the hexokinase gene significantly improved ATP content and this significantly improved contractility. These results demonstrate that elevating hexokinase activity can be beneficial to hypoxic or diabetic cardiomyocytes secondary to improving myocyte ATP levels. PMID- 16244375 TI - 3,4-Methylenedioxymethamphetamine activates nuclear factor-kappaB, increases intracellular calcium, and modulates gene transcription in rat heart cells. AB - 3,4-Methylenedioxymethamphetamine (MDMA) is an illicit psychoactive drug that has gained immense popularity among teenagers and young adults. The cardiovascular toxicological consequences of abusing this compound have not been fully characterized. The present study utilized a transient transfection/dual luciferase genetic reporter assay, fluorescence confocal microscopy, and gene expression macroarray technology to determine nuclear factor-kappaB (NF-kappaB) activity, intracellular calcium balance, mitochondrial depolarization, and gene transcription profiles, respectively, in cultured rat striated cardiac myocytes (H9c2) exposed to MDMA. At concentrations of 1 x 10(-3) M and 1 x 10(-2) M, MDMA significantly enhanced NF-kappaB reporter activity compared with 0 M (medium only) control. This response was mitigated by cotransfection with IkappaB for 1 x 10(-3) M but not 1 x 10(-2) M MDMA. MDMA significantly increased intracellular calcium at concentrations of 1 x 10(-3) M and 1 x 10(-2) M and caused mitochondrial depolarization at 1 x 10(-2) M. MDMA increased the transcription of genes that are considered to be biomarkers in cardiovascular disease and genes that respond to toxic insults. Selected gene activation was verified via temperature-gradient RT-PCR conducted with annealing temperatures ranging from 50 degrees C to 65 degrees C. Collectively, these results suggest that MDMA may be toxic to the heart through its ability to activate the myocardial NF-kappaB response, disrupt cytosolic calcium and mitochondrial homeostasis, and alter gene transcription. PMID- 16244376 TI - Direct implication of carbon monoxide in the development of heart failure in rats with cardiac hypertrophy subjected to air pollution. AB - Pollution is known to particularly affect patients with respiratory insufficiency and right ventricle abnormalities. We therefore hypothesized that carbon monoxide (CO) at low dose could be involved in cardiovascular disorders in patients with chronic hypoxic pulmonary hypertension secondary to chronic hypoxia. Ten-week-old male and female healthy Dark Agouti rats were randomly divided into two series- untrained (U) and trained (T)--of four groups of 18 animals each. Both U and T series were continuously exposed to ambient air (U(AIR), and T(AIR); n = 16) or air plus 50 ppm CO (U(AIR+CO) and T(AIR+CO); n = 18). Similarly, rats initially subjected to right ventricle hypertrophy secondary to chronic hypoxia (H) were continuously exposed to ambient air (TH(AIR), and UH(AIR); n = 18) or air plus 50 ppm CO (UH(AIR+CO), and TH(AIR+CO); n = 18). Doppler-echocardiography and hemodynamic studies performed at rest both indi-cated that CO had no significant effect on cardiac morphology or functions in control rats (U(AIR+CO) vs U(AIR)). In contrast, cardiac dilation and large decreases in left ventricular ejection fraction, mitral early diastolic rapid inflow (E) deceleration, E/atrial contraction filling (A) ratio, +dP/dt, and -dP/dt were found in TH(AIR+CO) compared with TH(AIR). After exposure, heart rate variability was unaffected in U(AIR+CO), whereas total power spectra were markedly decreased and low frequency/high frequency power ratio was increased in TH(AIR+CO) rats. CO pollution could be directly involved in cardiac disorders of patients with pre existent hypertrophic cardiomyopathies. PMID- 16244377 TI - Nitrate-based vasodilators inhibit multiple vascular aldehyde dehydrogenases. AB - Nitrate-based vasodilators (NBVs) are commonly used to treat multiple sequelae of atherosclerosis. A commonly used NBV, glyceryl trinitrate (GTN) is bioactivated by mitochondrial, class 2 aldehyde dehydrogenase (ALDH2). ALDH2 and other ALDHs are NAD(P)+-dependent enzymes critical to the detoxification of cytotoxic lipid aldehydes elevated in atherosclerotic lesions, such as trans-4-hydroxy-2-nonenal (HNE). The GTN bioactivation step, however, inac-tivates ALDH2 and may alter the metabolism of these aldehydes. In this study, we tested the hypothesis that multiple ALDH enzymes are inhibited by different NBVs. ALDH2, ALDH3A, and ALDH5A were present in aorta with ALDH2 and ALDH3A localized to the smooth muscle layers. GTN (1 microM) inhibited ALDH2 activity (55 +/- 6% of control) and ablated ALDH3 activity. In contrast, isosorbide-2,5-dinitrate (ISDN, 1 microM) inhibited ALDH3 activity (1.1 +/- 0.4% of control) but did not inhibit ALDH2 activity even up to 50 microM ISDN. In homogenates of rat aorta, GTN (1 microM) inhibited the NAD+-dependent (41 +/- 5% of control) and NADP+-dependent (25 +/- 6% of control) detoxification of HNE. The inhibition of ALDH3A, but not ALDH2, could be prevented by the addition of dithiothreitol. These studies demonstrate that GTN and ISDN possess selectivity for ALDH inactivation with different mechanisms of inactivation. PMID- 16244378 TI - Cardiac mitochondrial compromise in 1-yr-old Erythrocebus patas monkeys perinatally-exposed to nucleoside reverse transcriptase inhibitors. AB - Hearts from 1-yr-old Erythrocebus patas monkeys were examined after in utero and 6-wk-postbirth exposure to antiretroviral nucleoside reverse transcriptase inhibitors (NRTIs). Protocols were modeled on those given to human immunodeficiency virus (HIV)-1-infected pregnant women. NRTIs were administered daily to the dams for the last 20% or 50% of gestation, and to the infants for 6 wk after birth. Exposures included: no drug (n = 4); Zidovudine, 3'-azido-3' deoxythymidine (AZT; n = 4); AZT/Lamivudine, (-)-beta-L-2', 3'-Dideoxy-3' thiacytidine (Epivir, 3TC) (n = 4); AZT/Didanosine (Videx, ddI) (n = 4); and Stavudine (Zerit, d4T)/3TC (n = 4). Echocardiograms and clinical chemistry showed no drug-related changes, but the d4T/3TC-exposed fetuses at 6 and 12 mo had increased white cell counts (p < 0.05). At 1 yr of age, oxidative phosphorylation (OXPHOS) enzyme activities were similar in heart mitochondria from all groups. Mitochondrial pathology, that included clones of damaged mitochondria (p < 0.05), was found in hearts of all 1-yr drug-exposed infants. Levels of mtDNA were elevated (p < 0.05) in hearts of all NRTI-exposed monkeys in the following order: control < d4T/3TC < AZT < AZT/3TC < AZT/ddI. The clinical status of NRTI-exposed infants, as evidenced by behavior, clinical chemistry, OXPHOS activity and echocardiogram, was normal. However, extensive mitochondrial damage with clusters of similar-appearing damaged heart mitochondria observed by electron microscopy, and an increase in mtDNA quantity, that persisted at 1 yr of age, suggest the potential for cardiotoxicity later in life. PMID- 16244379 TI - Preparing for the next influenza pandemic. AB - The Dutch Ministry of Health, Welfare and Sport is determined to be as well prepared as possible for an influenza pandemic, if one should occur. Various measures based on the use of vaccines, antiviral agents and guidelines are being implemented or prepared. The aim is to maintain social and economic infrastructure as much as possible during a pandemic outbreak of influenza. In this context, the ministry is drawing on advice provided by the Health Council in two separate reports. PMID- 16244380 TI - Use of antiviral agents and other measures in an influenza pandemic. AB - The Dutch Ministry of Health asked the Health Council for advice on how to prepare for a possible influenza pandemic. In two advisory reports the Committee responsible indicated the measures that it believes would need to be taken if such a pandemic were to reach the Netherlands. During a pandemic, the Committee recommends that every resident of the Netherlands with influenza-like illness should be treated with neuraminidase inhibitors such as antiviral agents. This approach serves to mitigate the course of the disease, to reduce infectivity and to allow patients to build up immunity to the virus. Since up to 30% of the population could become ill, the Committee anticipates that a stock of five million courses of the neuraminidase inhibitor oseltamivir is sufficient. If a pandemic were to occur at a time that the stock does not exceed the present 225,000 courses, the committee advises restricting treatment to three specified groups of patients. If the first few patients are traced shortly after they fall ill, the Committee recommends treatment of the patient and postexposure prophylaxis for his/her close contacts. The Committee does not advocate prophylaxis in general, but it can envisage prophylaxis for particular groups of patients or under particular circumstances. The Committee believes that in order to reduce rapid spread of the virus, schools should be closed and events where large numbers of people gather in a confined space should be cancelled. Because this recommendation would have major social and economic consequences, the Committee understands that its implication will depend on the anticipated severity and extent of the pandemic. The Committee regards vaccination against influenza as the best means of protecting the population. The development of a vaccine should be the absolute priority. PMID- 16244381 TI - Helicobacter pylori, obesity and gastro-oesophageal reflux disease. Is there a relation? A personal view. AB - The incidence and prevalence of gastro-oesophageal reflux disease is rising. Changing dietary habits and increasing body weight can be held responsible. In several studies a close relation was found between body weight and the occurrence of reflux disease. It may be concluded that there is a definite relation between body mass index and the occurrence of reflux disease. H. pylori probably also plays a role. H. pylori causes changes in fundic leptin levels and plasma levels of ghrelin. Eradication of H. pylori infection can increase appetite leading to a rise in body mass index due to a higher caloric intake. H. pylori can be a 'protective' factor against the development of overweight. Since only a minority of overweight or obese patients with gastro-oesophageal reflux disease will lose weight successfully, medical treatment with effective acid suppression will be the mainstay of the treatment of reflux disease in patients with a high body mass index. PMID- 16244382 TI - Evaluation of endocrine tests. B: screening for hypercortisolism. AB - BACKGROUND: While reference values for 24-hour free urinary cortisol excretion and the overnight 1 mg dexa-methasone-suppression test in the healthy population are available, cut-off values in patients clinically suspected of Cushing's syndrome have to be established. METHODS: This was a prospective follow-up study in one academic centre of 144 patients with clinical suspicion of Cushing's syndrome (group A) and 50 patients with adrenal incidentaloma (group B) who were referred for putative hypercortisolism between 1 January 1993 and 1 January 2003. The 24-hour urinary free cortisol and post-dexamethasone plasma cortisol were measured. Accurate diagnosis of (absence of) Cushing's syndrome was confirmed by histopathological data and long-term follow-up. Based on the data obtained in group A, sensitivity, specificity and receiver operating characteristic (ROC) curves were calculated. RESULTS: Complete follow-up was obtained in 86%, and partial follow-up was obtained in 8% of patients. Median follow-up was 36 (1 to 122) months. In group A, 17 patients were found to have Cushing's syndrome. In this group median 24-hour urinary free cortisol was 77 (<5 to 51458) mmol/24 hours and median post-dexamethasone plasma cortisol was <50 (<50 to 4900) nmol/l. Area under the ROC curve was 0.958 for 24-hour urinary free cortisol and 0.985 for post-dexamethasone plasma cortisol. Optimal cut-off values were 180 nmol/24 hours (sensitivity 94%, specificity 94%) and 95 nmol/l (sensitivity 100%, specificity 94%) respectively. CONCLUSION: We established cut-off values for 24 hour free urinary cortisol excretion (180 nmol/24 hours) and for post dexamethasone plasma cortisol (95 nmol/l) in the evaluation of patients referred for hypercortisolism. PMID- 16244383 TI - Changes in antibiotic use in Dutch hospitals over a six-year period: 1997 to 2002. AB - OBJECTIVE: To analyse trends in antibiotic use in Dutch hospitals over the period 1997 to 2002. METHODS: Data on the use of antibiotics and hospital resource indicators were obtained by distributing a questionnaire to all Dutch hospital pharmacies. Antibiotic use was expressed as the number of defined daily doses (DDD) per 100 patient-days and as DDD per 100 admissions. RESULTS: Between 1997 and 2002, the mean length of stay decreased by 18%. The mean number of admissions remained almost constant. Total antibiotic use significantly increased by 24%, from 47.2 in 1997 to 58.5 DDD per 100 patient-days in 2002 (p<0.01), whereas expressed as DDD per admissions it remained constant. Antibiotic use varied greatly between the hospitals. Moreover, the mean number of DDD per hospital of amoxicillin with clavulanic acid, clarithromycin, cefazolin, clindamycin and ciprofloxacin increased by 16, 38, 39, 50 and 52%, respectively. Total antibiotic use was higher in university hospitals than in general hospitals. CONCLUSIONS: Between 1997 and 2002, patients hospitalised in the Netherlands did not receive more antibiotics but, since they remained in the hospital for fewer days, the number of DDD per 100 patient-days increased. For macrolides, lincosamides and fluoroquinolones increases in both DDD per 100 patient-days and in DDD per 100 admissions were observed. It is arguable whether these trends result in an increase in selection pressure towards resistance in the hospitals. Continuous surveillance of antibiotic use and resistance is warranted to maintain efficacy and safety of antibiotic treatment. PMID- 16244384 TI - Acute lymphoblastic leukaemia in pregnancy. AB - Two cases of pregnant patients with acute lymphoblastic leukaemia (ALL) are presented. ALL is rare in pregnancy. The basic principle of ALL treatment is combination chemotherapy with sequential administration of induction, consolidation and maintenance therapy, and this also holds for ALL in pregnancy. The prognosis of ALL in pregnancy is poor and termination of the pregnancy needs to be considered. PMID- 16244385 TI - Early onset of oral aphthous ulcers with weekly docetaxel. AB - Two patients with metastatic breast cancer developed oral aphthous ulcers after only two administrations of weekly docetaxel without any other toxicity. A treatment delay and dose reduction appears to be an effective management strategy. PMID- 16244386 TI - Fever from the USA or Portugal? PMID- 16244387 TI - Dutch medical oath. AB - In the first part of this article, the booklet Dutch Medical Oath is reviewed. The content of the new oath is discussed as are the reasons for revision of the previous version of the oath. This is followed by a short history of the oath. In the second part of the article the oath is compared with the seven competencies of a medical specialist. The new oath contains elements of six of these seven competencies. This demonstrates that the oath is in keeping with the new medical educational demands. PMID- 16244389 TI - Development of methods for the quantification of essential and toxic elements in human biomonitoring. AB - Analytical methods using sector field ICP-MS and ICP-AES were developed for the determination of Al, Ba, Be, Bi, Ca, Cd, Co, Cr, Cu, Fe, Hg, Li, Mg, Mn, Mo, Ni, Pb, Sb, Si, Sn, Sr, Tl, V, W, Zn and Zr in cerebrospinal fluid, urine, serum and blood. Sample treatment procedures merging high sample throughput, simplicity and low contamination risk were set up. Method performances were evaluated in terms of detection limits, accuracy and precision. The limits were below 0.05 ng ml(-1) for all the elements, except for Al (all matrices), Hg (blood), Pb (blood) and Sn (serum and blood). The accuracy varied from 86% to 110% and the precision was always below 6%. PMID- 16244388 TI - Metal ions and oxidative protein modification in neurological disease. AB - This review highlights the role of oxidative stress and imbalances in metal ion homeostasis in the neurodegenerative diseases Alzheimer's disease and Parkinson's disease and in the progressive demyelinating disease multiple sclerosis. The chemistry and biochemistry of oxidative stress-induced protein damage are first described, followed by the evidence for a pathological role of oxidative stress in these disease states. It is tempting to speculate that free radical oxygen chemistry contributes to pathogenesis in all these conditions, though it is as yet undetermined what types of oxidative changes occur early in the disease, and what types are secondary manifestations of neuronal degeneration. PMID- 16244390 TI - The network of the Italian laboratories: a proficiency test on the quantification of trace elements in serum. AB - A proficiency test on the quantification of Al, Cu, Mn, Se and Zn in serum was carried out to verify the performance of about 30 regional laboratories of the network of Italian laboratories. The exercise consisted of four runs in which the laboratories were free in choosing analytical methods to determine trace elements in freeze-dried animal serum. Laboratories performances were evaluated by the study of statistical functions as Coefficients of Variation (CV), Youden plot and Z-score value. As for Al, the results were generally characterized by good accuracy and precision, in spite of the low levels of the element (5-7 microg l( 1)). Copper determination had some problems only at low concentration (about 160 microg l(-1)--first run), in which an elevated number of anomalous data were found. Better data were achieved for Zn, for which anomalous values were mainly stragglers than outliers. Due to the low number of data for Mn (concentrations from 0.6 to 60 microg l(-1)) and Se (concentration from 45 to 106 microg l(-1)), a restricted statistical treatment was applied; for these elements high CV values were found (range from 30 to 80%). The results of this trial confirmed that participation in a proficiency test represents a benefit for all analytical laboratories. In fact, with few exceptions, most of the participants improved their performances in terms of Z-score values. PMID- 16244391 TI - Assessment of reference values for selected elements in a healthy urban population. AB - Reference values for 26 elements, namely Al, Ba, Be, Bi, Ca, Cd, Co, Cr, Cu, Fe, Hg, Li, Mg, Mn, Mo, Ni, Pb, Sb, Si, Sn, Sr, Tl, V, W, Zn and Zr are proposed in serum and blood of 110 healthy adults of the urban area of Rome. They were included in the study on the basis of strict criteria of eligibility and exclusion. With the exception of Ba, Bi, Co, Cr, Ni, Sb, Sn Tl in serum, and Bi, Hg, Si, V and W in whole blood, experimental data for each all the other analytes were found to approach a normal distribution. The estimated 5-95% references ranges (in ng ml(-1)) were reported. For several elements the reference ranges observed overlapped information available in the literature. Gender, age, body mass index, smoking habits and alcohol consume were used as grouping variables. Mutual associations were observed for several elements, as follows: Be, Ca, Co, Cr, Cu, Li, Mo, Pb and Zn with sex; Ca, Pb and Si with age (< and > 45 years); Co, Cr, Mo, Sb and Tl with body mass index; Cd and Pb with smoking habit; Cr and Pb with alcohol consume. PMID- 16244392 TI - Metals and oxidative stress in patients with Parkinson's disease. AB - Twenty-six metals and the oxidative status in 71 patients affected by Parkinson's disease and 44 healthy individuals were compared in order to identify potential biomarkers of the disease. In the patients, the following significant imbalances were found (p < or = 0.05): i) in serum, an increment of Ca, Mg, Ni, Si and V, and a decrement of Cd, Co, Fe, Li, Sn, Zn and Zr; ii) in blood, raised levels of Co, Li, Ni and Si and decreased of Al, Be, Ca, Cd, Fe, Mg, Mo, Sn, Zn and Zr; iii) increased formation of oxidant species and lowered anti-oxidant capacity (p < or = 0.001 for both). Barium, Bi, Cr, Cu, Hg, Mn, Pb, Sb, Sr, Tl and W did not change with the disease. The best discriminating variables between patients and controls were Cd, Co, Fe, Ni and Si in serum (91.2% of cases correctly classified), and Al, Cd, Co, Fe, Mo and Si in blood (98.2% of cases properly classified). PMID- 16244393 TI - Monitoring of chemical elements and oxidative damage in patients affected by Alzheimer's disease. AB - The haematic concentration of 26 metals and the oxidative damage in 60 patients (20 males and 40 females) affected by Alzheimer's disease and 44 healthy individuals (33 males and 11 females) were compared. In patients, the following significant (p < or = 0.05) discrepancies were found: i) increment of Ca, Cd, Hg, Mg, Si and Sn, and decrement of Al, Co, Fe and Zn in serum; ii) higher concentrations of Cu, Li, Mn, Sn and Zr and lower of Fe, Hg, Mo in blood; iii) overproduction of oxidant species (SOS) and decrease of the anti-oxidant capacity (SAC) (p < or = 0.001, for both). Variables that, joined, better discriminated between patients and controls resulted to be Si, SOS, SAC, Co, Ca, Al in serum (94% of cases correctly classified) and Cu, Zr, Mo and Fe in blood (90% of cases properly categorized). PMID- 16244394 TI - Correlation between metal ions and clinical findings in subjects affected by Alzheimer's disease. AB - There is a growing interest to evaluate metals in biological fluids in Alzheimer's disease (AD). There are numerous studies on this theme, but just few papers analyzed the relationship between haematic metal concentrations and the clinical features of the disease. In this study, possible associations between clinical features of AD and the variations in serum and blood concentration of some metals, as well as the serum oxidative status and the antioxidant capacity have been investigated. Sixty subjects with AD were enrolled. Some elements correlated with gender, depression and duration of the disease. However, the most significant result was the relationship between blood Ca and Fe levels and the severity of cognitive impairment. We hypothesize that Ca and Fe might play an important role in the pathogenetic mechanisms of AD. PMID- 16244395 TI - Quantification of chemical elements in blood of patients affected by multiple sclerosis. AB - Although some studies suggested a link between exposure to trace elements and development of multiple sclerosis (MS), clear information on their role in the aetiology of MS is still lacking. In this study the concentrations of Al, Ba, Be, Bi, Ca, Cd, Co, Cr, Cu, Fe, Hg, Li, Mg, Mn, Mo, Ni, Pb, Sb, Si, Sn, Sr, Tl, V, W, Zn and Zr were determined in the blood of 60 patients with MS and 60 controls. Quantifications were performed by inductively coupled plasma (ICP) atomic emission spectrometry and sector field ICP mass spectrometry. When the two groups were compared, an increased level of Co, Cu and Ni and a decrement of Be, Fe, Hg, Mg, Mo, Pb and Zn in blood of patients were observed. In addition, the discriminant analysis pointed out that Cu, Be, Hg, Co and Mo were able to discriminate between MS patients and controls (92.5% of cases correctly classified). PMID- 16244396 TI - Concentration of elements in serum of patients affected by multiple sclerosis with first demyelinating episode: a six-month longitudinal follow-up study. AB - Twenty-six chemical elements and oxidative status were determined in serum of 12 patients with first demyelinating episode and brain magnetic resonance imaging compatible with the disease at different time points. Quantifications of Al, Ba, Be, Bi, Ca, Cd, Co, Cr, Cu, Fe, Hg, Li, Mg, Mn, Mo, Ni, Pb, Sb, Si, Sn, Sr, V, Tl, W, Zn and Zr, as well as of serum oxidative status and antioxidant capacity were carried out. The results were compared with values obtained from healthy subjects living in the same geographic area. Concentration variability, expressed as coefficient of variation (CV), was evaluated over a six months longitudinal follow-up. The CV was higher for Li and Pb, while showed minimal variation for Ca, Cu, Mg and Zn--elements strictly body regulated. Significant difference (p < or = 0.05) in mean concentrations of Ba, Ca, Cd, Cr, Li, Mn, Mo, Ni, Sb, Si, Sn and Zr between patients at time 0 and controls was also found. PMID- 16244397 TI - Discriminant analysis to study trace elements in biomonitoring: an application on neurodegenerative diseases. AB - Quantification of 26 elements was performed in blood of patients affected by neurodegenerative pathologies, i.e., Alzheimer's disease (AD), Parkinson's disease (PD) and multiple sclerosis (MS), and of a control group to study the potential role of blood elements as markers for the different neurodegenerations. A multivariate discriminant analysis (stepwise method) was applied to determine the best set of variables to discriminate among subjects with different health status. Preliminary results show three classification functions of seven elements, namely Ca, Co, Cu, Fe, Ni, Pb and Zr. PMID- 16244398 TI - Brain damage in preterm infants: etiological pathways. AB - Preterm newborns represent a high-risk population for brain damage, primarily affecting the white matter, and for related neurodevelopmental disabilities. Determinants of brain damage have been extensively investigated, but there are still many controversies on how these factors can influence the developing brain and provoke damage. The concept of etiological pathway, instead of a single determinant, appears to better explain pathogenetic mechanisms: the brain damage may represent the final outcome of exposure to several combinations of risk factors in the same pathway or in different pathways and can change according to the gestational age. The aim of this article is to review the current knowledge on the pathogenesis of brain damage in preterm infants, within the frame of two main theoretical models, the ischemic and the inflammatory pathway. The relationship between the two pathways and the contribution of genetic susceptibility to ischemic and/or inflammatory insult, in modulating the extent and severity of brain damage, is also discussed. PMID- 16244399 TI - [Prevalence of the use of old and new drugs among new entrants in Italian prisons]. AB - An epidemiological survey was conducted to evaluate the prevalence of drug use among new entrants in Italian prisons. Overall, 1267 inmates were recruited by 9 prisons. Drug use before imprisonment was reported by 55.8% of the participants. Cocaine was the most commonly used drug (42%), followed by heroin (34%), marijuana/cannabis (33%), ecstasy (7%), hallucinogens (6%), amphetamines (5%); more than one drug was reported by 68% of abusers. Recent use (1 month before imprisonment) was admitted by up to 27% of inmates. Alcohol or tobacco use was reported by 38 and 77% of the inmates, respectively. Our findings indicate that a high proportion of inmates has ever used drugs; adequate intervention is needed to reduce the risk of addictive behaviour in this population group. PMID- 16244400 TI - Laboratory study to investigate the efficacy of cyfluthrin EW treated nets against phlebotomine sandflies. AB - Toxic and feeding inhibition effect of cyfluthrin treated nets was assessed and compared with the activity of deltamethrin impregnated nets, before and after 2 and 4 washes against Phlebotomus papatasi. Impregnated cyfluthrin nets (50, 25, 12.5 mg active ingredient (a.i.)/m2) caused knock down and mortality rates higher than 90%, being of 100% at the highest dose, while the nets impregnated with deltamethrin (25, 12.5, 6.25 mg a.i./m2) showed low mortality rates (13.3-43.5%). Also after washing cyfluthrin at the dose of 50 mg a.i./m2 provided higher activity than deltamethrin (25 mg a.i./m2), being mortality rates within 24 hs of 66.7 and 48.9% for cyfluthrin and 38.0 and 2.1% for deltamethrin, respectively after 2 and 4 washes. Before washing, cyfluthrin (50 mg a.i./m2) significantly prevented blood feeding, with respect to both untreated (88.6%) and deltamethrin impregnated nets (56.8%). Deltamethrin (25 mg a.i./m2) showed low feeding inhibition (48.4%), although it was significantly higher than control. The washing of cyfluthrin impregnated nets (50 mg a.i./m2) produced a decrease of the feeding inhibition, being the reduction of 26.6% and 41.3%, respectively after 2 and 4 washes. Two washes of deltamethrin impregnated nets (25 mg a.i./m2) yielded a very high reduction of antifeeding inhibition (68.8%). These laboratory results if confirmed by field evaluation indicate cyfluthrin as good candidate for treatment of mosquito nets to be used against phlebotomine sandflies. PMID- 16244401 TI - [Evaluation of repellent and anti-feeding effect of garlic oil (Allium sativum) against the bite of phlebotomine sandflies Diptera: Psychodidae]. AB - The repellent and anti-feeding effect of garlic oil was evaluated in laboratory conditions against the bite of Phlebotomus papatasi females. The effectiveness was evaluated by two different laboratory procedures: (i) topical application of garlic oil on five human volunteers, using the "standard cage test", and (ii) feeding sandflies on artificial membranes treated with the compound. Garlic oil showed a significant protection by topical application on the skin of volunteers, being the protection 97% and 40%, respectively at 1% and 0.005% dilution. Garlic oil showed also an anti-feeding effect when tested on the artificial membrane feeding apparatus; the anti-feeding effect was dose dependent, being 100% at 1%. The results of the present study confirm previous observations on the repellent effect of garlic oil against haematophagous arthropods. PMID- 16244402 TI - Double-helix structure in multiwall boron nitride nanotubes. AB - A new nanotube structural form is reported that resembles a double helix in multiwall boron nitride nanotubes (MW-BNNT) grown by a carbon-free chemical-vapor deposition process as documented by evidence obtained by transmission electron diffraction and microscopy. The double-helix structure is found in MW-BNNTs exhibiting the same chirality in its different walls. The MW-BNNTs deviate from the structure of ideal nested coaxial cylindrical tubes. Most significantly, bright- and dark-field electron imaging reveals regular zigzag dark and bright spots on the side walls of the nanotubes. The repeating distance between the bright, or dark, spots is related to the chiral angle of the nanotube. Electron diffraction patterns recorded from individual nanotubes show additional diffraction spots belonging to the 201 zone axes, which are not allowed in a perfectly cylindrical nanotube. These additional diffraction spots become asymmetrical as smaller sections of the nanotube are probed. A series of diffraction patterns recorded along the tube axis showed that the imperfections giving rise to these spots move in a regular fashion around the circumference of the tube. It is shown that all experimental evidence supports the structure model of two helices; one is polygonal in cross section and highly crystalline and the other is circular and less ordered. It is further suggested that the double-helix structure is a result of stronger wall-wall interactions associated with the ionic bonding in boron nitride. PMID- 16244403 TI - Combinatorial construction of tilings by barycentric simplex orbits (D symbols) and their realizations in Euclidean and other homogeneous spaces. AB - A new method, developed in previous works by the author (partly with co-authors), is presented which decides algorithmically, in principle by computer, whether a combinatorial space tiling (Tau, Gamma) is realizable in the d-dimensional Euclidean space E(d) (think of d = 2, 3, 4) or in other homogeneous spaces, e.g. in Thurston's 3-geometries: E(3), S(3), H(3), S(2) x R, H(2) x R, SL(2)R, Nil, Sol. Then our group Gamma will be an isometry group of a projective metric 3 sphere PiS(3) (R, < , >), acting discontinuously on its above tiling Tau. The method is illustrated by a plane example and by the well known rhombohedron tiling (Tau, Gamma), where Gamma = R3m is the Euclidean space group No. 166 in International Tables for Crystallography. PMID- 16244405 TI - Ultra-small-angle X-ray scattering from dislocation structures. AB - Ultra-small-angle X-ray scattering data were obtained from deformed single crystal aluminium samples. These data are consistent with recent theoretical predictions of scattering from dislocation walls, allowing quantitative microstructural parameters to be extracted. PMID- 16244404 TI - Anomalous scattering factor determined by semicircle fitting near the K absorption edge of Ge. AB - It is shown that the locus of the f' + if'' plot in the complex plane, f' being determined from measured f'' by using the dispersion relation, looks like a semicircle very near the absorption edge of Ge. The semicircular locus is derived from a quantum theory of X-ray resonant scattering when there is a sharp isolated peak in f'' just above the K-absorption edge. Using the semicircular behavior, an approach is proposed to determine the anomalous scattering factors in a crystal by fitting known calculated values based on an isolated-atom model to a semicircular focus. The determined anomalous scattering factors f' show excellent agreement with the measured values just below the absorption edge. In addition, the phase determination of a crystal structure factor has been considered by using the semicircular behavior. PMID- 16244406 TI - Abinitio structure determination using dispersive differences from multiple wavelength synchrotron-radiation powder diffraction data. AB - The purpose of this paper and a test case study is to assess a method of ab initio structure solution from powder diffraction data using f' difference techniques. A theoretical foundation for the approach used is first provided. Then, with a test case (nickel sulfate hexahydrate), it is shown that both the position of the anomalous scatterer (Ni) can be determined and the structure can be developed in full. Specifically, synchrotron-radiation data were collected at two wavelengths close to the K edge for Ni and three wavelengths remote from the Ni absorption edge, at 1.3, 1.8 and 2.16 A. These five wavelengths then allowed various combinations to be tried to establish which wavelength pairs gave the optimum signal in the Patterson maps using dispersive amplitude differences. The initial phases derived from the metal-atom position then allowed the structure to be fully developed by difference Fourier cycling. The relevance of these developments to structure-solution possibilities for proteins via powder dispersive difference data is then outlined. PMID- 16244407 TI - Report of the working group on crystal phase identifiers. AB - The proposed crystalline phase identifier consists of a number of components (layers) describing enough properties of the phase to allow a unique identification. These layers consist of the chemical formula, a flag indicating the state of matter, the space-group number and the Wyckoff sequence. They are defined in such a way that they can be incorporated into the IUPAC International Chemical Identifier (InChI) proposed by the International Union of Pure and Applied Chemistry (IUPAC). PMID- 16244408 TI - The variety of convex 12-hedra revised. AB - The symmetry point group statistics for all combinatorially non-isomorphic convex 12-hedra (6384634 in total) are contributed in the paper. The numbers of 12-hedra with 11 to 19 vertices are revised. The most symmetrical shapes with 6 to 120 automorphism group orders (115 in total) are drawn in Schlegel diagrams and characterized by facet symbols and symmetry point groups. PMID- 16244409 TI - On the Motzkin-Grunbaum theorem. AB - A constructive proof of the Motzkin-Grunbaum theorem (at least one fullerene C(n) with any even n > or = 24 exists) is suggested. PMID- 16244412 TI - Mini-implants: Where are we? PMID- 16244414 TI - Non-surgical treatment of an adult skeletal Class III patient with insufficient incisor display. PMID- 16244415 TI - A mini-maxillary protractor for Class III correction. PMID- 16244416 TI - The readers' corner. How often do you offer limited treatment? PMID- 16244417 TI - The readers' corner. What is your usual appointment interval? PMID- 16244418 TI - The Greenfield lingual distalizer. PMID- 16244419 TI - Fixed space maintainer for use with a rapid palatal expander. PMID- 16244420 TI - Correction of a malocclusion caused by a dentigerous cyst. PMID- 16244421 TI - Treatment of ectopic maxillary canines using a palatal implant for anchorage. PMID- 16244422 TI - Quick setup for light-cured bonding. PMID- 16244423 TI - A removable Class II appliance for simultaneous distalization and expansion. PMID- 16244424 TI - An American in Paris. PMID- 16244425 TI - A new self-etching, light-cured bonding system. PMID- 16244427 TI - Six steps to wealth and financial independence. PMID- 16244428 TI - Effect of randomly interesterified triacylglycerols containing medium- and long chain fatty acids on energy expenditure and hepatic fatty acid metabolism in rats. AB - In our previous studies, medium- and long-chain triacylglycerols (MLCT), randomly interesterified triacylglycerols containing medium-chain and long-chain fatty acids in the same glycerol molecule, significantly reduced body fat accumulation in humans and rats. To clarify mechanism(s) for this effect of MLCT, we measured energy expenditure and hepatic fatty acid metabolism in rats by comparison with long-chain triacylglycerols (LCT) or medium-chain triacylglycerols (MCT). MLCT, compared with LCT, showed significantly lower body fat accumulation, higher 24-h energy expenditure and acyl-CoA dehydrogenase activity measured using octanoyl CoA as a substrate, and similar lipogenic activity. MCT, compared with LCT, showed significantly higher energy expenditure, but fat accumulation was comparable. Additionally, MCT exhibited significantly higher lipogenic activity than the other oils. These data suggest that enhancement of energy expenditure and medium-chain fatty acids (MCFA) oxidation without activating de novo lipogenesis are responsible at least for the lower body fat accumulation in rats fed MLCT. The activation of hepatic lipogenesis by excessive intake of MCFA might counteract their preventive effects on body fat accumulation. PMID- 16244426 TI - An adjustable surgical guide for miniscrew placement. PMID- 16244429 TI - Attenuated response of the serum triglyceride concentration to ingestion of a chocolate containing polydextrose and lactitol in place of sugar. AB - We examined the effects of ingesting a non-sugar chocolate containing polydextrose and lactitol in place of sucrose and lactose on the concentrations of plasma glucose and serum insulin and triglyceride in humans. A regular chocolate was used as the control. A crossover study was employed, and the subjects each ingested 46 g of the control or non-sugar chocolate in the experiments. Alterations in the blood components were monitored for a period of 150 min after ingestion. The control chocolate elevated the concentrations of plasma glucose and serum insulin, with the peak occurring 30 min after ingestion, but the non-sugar chocolate had a very minor effect. The serum triglyceride concentration gradually increased after ingesting the control chocolate, but was only slightly elevated 150 min after ingesting the non-sugar chocolate. An animal study also showed an attenuated response of serum triglyceride to the administration of a fat emulsion containing polydextrose and lactitol, suggesting that the triglyceride transit through the gut was promoted by these compounds. These results suggest that, compared to regular chocolate, fat absorption in the gut was less after ingesting the non-sugar chocolate, presumably resulting in less effect on body fat deposition. PMID- 16244430 TI - Inhibitory effect of porphyran, prepared from dried "Nori", on contact hypersensitivity in mice. AB - Porphyran is a major component of the red algae, Porphyra tenera and P. yezoensis, which are processed into a sheet type of dried food, "Nori". Porphyran has been reported to activate murine macrophages by in vitro and i.p. injection studies. The contact hypersensitivity (CHS) reaction in mice is commonly used as a model to evaluate the anti-allergic activity of food and food components. We therefore studied the effect of porphyran on the CHS reaction in Balb/c mice to evaluate anti-allergic activity of porphyran. We found that an oral administration of porphyran (2% in drinking water) suppressed the CHS reaction (ear edema) induced by 2,4,6-trinitrochlorobenzene. We also found that porphyran suppressed the serum level of IgE and the production of interferon-gamma (IFN gamma) in the challenged ear lobe. We conclude from these results that the CHS reaction was suppressed by oral porphyran due to the decreased serum level of IgE and the production of IFN-gamma in the challenged ear lobe. PMID- 16244431 TI - Ovipositional deterrent in the sweet pepper, Capsicum annuum, at the mature stage against Liriomyza trifolii (Burgess). AB - Liriomyza trifolii (Burgess), the American serpentine leafminer fly, is well known as a serious pest throughout the world. This insect attack over 21 different plant families including solanaceae plants. The mature sweet pepper, Capsicum annuum (Solanaceae), however, shows resistance to this leafminer fly. This resistance is based on the ovipositional deterrent in the sweet pepper leaf against the fly species. Based on bioassay-guided fractionation, luteolin 7-O beta-D-apiofuranosyl-(1-->2)-beta-D-glucopyranoside was isolated and identified as the ovipositional deterrent against this insect species. This compound completely deterred L. trifolii females from laying their eggs on a host plant leaf treated at 4.90 microg/cm2. PMID- 16244433 TI - Corn silk induced cyclooxygenase-2 in murine macrophages. AB - Stimulation of murine macrophages with corn silk induced cyclooxygenase (COX)-2 with secretion of PGE2. Expression of COX-2 was inhibited by pyrolidine dithiocarbamate (PDTC), and increased DNA binding by nuclear factor kappa B (NF kappaB), indicating that COX-2 induction proceeds also via the NF-kappaB signaling pathway. A specific inhibitor of COX-2 decreased the expression level of inducible nitric oxide synthase (iNOS) stimulated by corn silk. PGE2 elevated the expression level of iNOS, probably via EP2 and EP4 receptors on the surface of the macrophages. PMID- 16244432 TI - Distribution of PCDDs/PCDFs and Co-PCBs in human maternal blood, cord blood, placenta, milk, and adipose tissue: dioxins showing high toxic equivalency factor accumulate in the placenta. AB - To assess levels of dioxin background contamination and transfer of dioxins from mothers to unborn children and infants, concentrations of polychlorinated dibenzo p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and coplanar polychlorinated biphenyls (Co-PCBs) were measured in human samples from expectant and nursing mothers living in Nara, Japan. The average toxic equivalency quantities (TEQs) of PCDDs/PCDFs and Co-PCBs from circulating maternal blood, cord blood, placenta, milk taken 3-10 d after delivery, milk taken one month after delivery, and adipose tissue were 26 and 9.3, 15 and 2.3, 31 and 1.2, 16 and 5.4, 18 and 8.8, and 16 and 7.7 pg-TEQ/g-fat, respectively. Among the various PCDD/PCDF congeners, 1,2,3,7,8-PeCDD and 2,3,4,7,8-PeCDF contributed most heavily to the TEQs of all maternal samples. Among the various Co-PCB congeners, 3,3',4,4',5-PeCB (#126), 2,3,3',4,4',5-HxCB (#156), and 2,3',4,4',5-PeCB (#118) contributed most heavily to the TEQs of all maternal samples. But, the concentrations and relative percentages of congeners differed among the various samples, suggesting that congeners showing high toxic equivalency factor accumulate in the placenta. PMID- 16244434 TI - New deblocking aminopeptidases from Pyrococcus horikoshii. AB - It has been reported that one of the hyperthermostable aminopeptidases from Pyrococcus horikoshii exhibits hydrolytic activity toward short peptides and acyl peptides (deblocking activity). In the genome database of P. horikoshii, two new open reading frames homologous to the hyperthermostable aminopeptidase of P. horikoshii were found. The two new genes for the proteins were cloned, expressed using E. coli, and characterized. The purified proteins gave a single band on SDS PAGE corresponding to molecular masses of 42 kDa and 41 kDa respectively, and exhibited aminopeptidase activity, including deblocking activity. These enzymes are likely to exist as oligomeric structures at neutral pH. The optimum pHs of the two enzyme activities were in the range of 7.0 to 7.5, and the optimum temperatures for the activities were around 100 degrees C. The enzymes exhibited low hydrolytic activity for peptide substrates longer than 10 residues. They were activated by cobalt and zinc ions. Their substrate specificities and activation factors are different. It was confirmed that P. horikoshii has three similar aminopeptidases with deblocking activity and that these enzymes appear to play important roles in hydrolyzing small peptides in P. horikoshii cells. PMID- 16244435 TI - Molecular properties and enhancement of thermostability by random mutagenesis of glutamate dehydrogenase from Bacillus subtilis. AB - The rocG gene encoding glutamate dehydrogenase from Bacillus subtilis (Bs-GluDH) was cloned, and expressed at considerable magnitude in Escherichia coli. The recombinant Bs-GluDH was purified to homogeneity and has been determined to have a hexameric structure (M(r) 270 kDa) with strict specificity for 2-oxoglutarate and L-glutamate, requiring NADH and NAD+ as cofactors respectively. The enzyme showed low thermostability with T(m) = 41 degrees C due to dissociation of the hexamer. To improve the thermostability of this enzyme, we performed error-prone PCR, introducing random mutagenesis on cloned GluDH. Two single mutant enzymes, Q144R and E27F, were isolated from the final mutant library. Their T(m) values were 61 degrees C and 49 degrees C respectively. Furthermore, Q144R had a remarkably high k(cat) value (435 s(-1)) for amination reaction at 37 degrees C, 1.3 times higher than that of the wild-type. Thus, Q144R can be used as a template gene to modify the substrate specificity of Bs-GluDH for industrial use. PMID- 16244436 TI - Expression level of ABCG2 in the placenta decreases from the mid stage to the end of gestation. AB - The aim of this study was to elucidate the expression pattern of ABCG2 in the placenta from the mid stage to the end of gestation. rABCG2 expression was investigated in rats on the 14th gestation day (gd) and the 20th gd. Expression of the rABCG2 gene and expression of rABCG2 protein in the placenta were detected on gd 14 by RT-PCR and Western blot analysis respectively. The expression level of rABCG2 on gd 20 was less than that on gd 14. We investigated whether progesterone, secreted from the placenta, regulates the expression of ABCG2 in BeWo cells. Expression levels of the ABCG2 gene and protein in BeWo cells were decreased by progesterone treatment. We conclude that progesterone plays a role in reduction in the expression level of ABCG2 in the placenta with the advance of gestation from the mid stage to the end of gestation. PMID- 16244437 TI - Degradation of soluble proteins including some allergens in brown rice grains by endogenous proteolytic activity during germination and heat-processing. AB - The effect of germination and subsequent heat-processing on the degradation of soluble proteins, including some allergenic proteins, in brown rice grains was investigated. The content of soluble proteins, including 14-16-kDa and 26-kDa allergens, in the germinated and processed brown rice grains (GPR) was much lower than that of non-germinated brown rice. These proteins in brown rice grains were also much lower after subsequent heat-processing during the manufacturing process. The protease activity of germinated brown rice (GR) was detected and increased 1.5 times after germination. The optimum pH values for degradation of the 26-kDa and 14-16-kDa allergens in the GR grains were 4 and between 5 and 7, respectively. These results suggest that the decrease in the soluble proteins and allergens was induced in part by proteolytic degradation. The presence of a detergent enhanced the proteolytic degradation of the soluble proteins, especially of the 26-kDa allergen, in the brown rice grains. The degradation of the 26-kDa allergen was weakly inhibited by NEM, suggesting cysteine protease(s) may have been involved in its degradation. These results suggest that the two abundant allergens were degraded in a different manner and probably by different proteases in the grains during germination. PMID- 16244439 TI - Efficient route to (S)-azetidine-2-carboxylic acid. AB - A new and efficient route to (S)-azetidine-2-carboxylic acid (>99.9% ee) in five steps and total yield of 48% via malonic ester intermediates was established. As the key step, efficient four-membered ring formation (99%) was achieved from dimethyl (S)-(1'-methyl)benzylaminomalonate by treating with 1,2-dibromoethane (1.5 eq) and cesium carbonate (2 eq) in DMF. Krapcho dealkoxycarbonylation of dimethyl (1'S)-1-(1'-methyl)benzylazetidine-2,2-dicarboxylate, the product of this cyclization procedure, proceeded with preferential formation (2.7:1, 78% total yield) of the desired (2S,1'S)-monoester, with the help of a chiral auxiliary which was introduced on the nitrogen atom. The undesired (2R,1'S) isomer could be converted to that with proper stereochemistry, by a deprotonation and subsequent re-protonation step. Finally, lipase-catalyzed preferential hydrolysis of the (2S,1'S)-monoester and subsequent deprotection provided enantiomerically pure (S)-azetidine-2-carboxylic acid in a 91% yield from the mixture of (2S,1'S)- and (2R,1'S)-isomers. PMID- 16244438 TI - Purification and biochemical characterization of the F1-ATPase from Acidithiobacillus ferrooxidans NASF-1 and analysis of the atp operon. AB - ATPase was purified 51-fold from a chemoautotrophic, obligately acidophilic iron oxidizing bacterium, Acidithiobacillus ferrooxidans NASF-1. The purified ATPase showed the typical subunit pattern of the F1-ATPase on a polyacrylamide gel containing sodium dodecyl sulfate, with 5 subunits of apparent molecular masses of 55, 50, 33, 20, and 18 kDa. The enzyme hydrolyzed ATP, GTP, and ITP, but neither UTP nor ADP. The K(m) value for ATP was 1.8 mM. ATPase activity was optimum at pH 8.5 at 45 degrees C, and was activated by sulfite. Azide strongly inhibited the enzyme activity, whereas the enzyme was relatively resistant to vanadate, nitrate, and N,N'-dicyclohexylcarbodiimide. The genes encoding the subunits for the F1F(O)-ATPase from A. ferrooxidans NASF-1 were cloned as three overlapping fragments by PCR cloning and sequenced. The molecular masses of the alpha, beta, gamma, delta, and epsilon subunits of the F1 portion were deduced from the amino acid sequences to be 55.5, 50.5, 33.1, 19.2, and 15.1 kDa, respectively. PMID- 16244440 TI - Novel application of Shochu distillery by-products to prophylaxis against mammary carcinogenesis induced by 7,12-dimethylbenz[a]anthracene in rats. AB - The effect of the heat-dried product of Shochu distillery by-products (HSDB) derived from sweet potato on mammary carcinogenesis in rats was investigated. HSDB was fed at 2.5% or 5% of the total feed weight. Dietary HSDB at the 5% level suppressed the incidence and number of tumors, and delayed the latency of mammary tumor development relative to the control diet. Experiments were conducted to determine the relative polarity of the anticarcinogenic constituent(s). The number of tumors per tumor-bearing rat was lower in the diet group fed with an ethyl acetate extract of HSDB than in the control group. The tumor incidence evaluated at both palpation and autopsy was slightly lower in the group fed with a methanol extract than in the control group. These results suggest that HSDB contained at least two constituents of differing polarity that counteracted mammary carcinogenesis. PMID- 16244442 TI - Purification and characterization of a novel aminoacylase from Streptomyces mobaraensis. AB - A novel aminoacylase was purified to homogeneity from culture broth of Streptomyces mobaraensis, as evidenced by SDS-polyacrylamide gel electrophoresis (PAGE). The enzyme was a monomer with an approximate molecular mass of 100 kDa. The purified enzyme was inhibited by the presence of 1,10-phenanthroline and activated by the addition of Co2+. It was stable at temperatures of up to 60 degrees C for 1 h at pH 7.2. It showed broad substrate specificity to N acetylated L-amino acids. It catalyzed the hydrolysis of the amide bonds of various N-acetylated L-amino acids, except for Nepsilon-acetyl-L-lysine and N acetyl-L-proline. Hydrolysis of N-acetyl-L-methionine and N-acetyl-L-histidine followed Michaelis-Menten kinetics with K(m) values of 1.3+/-0.1 mM and 2.7+/-0.1 mM respectively. The enzyme also catalyzed the deacetylation of 7 aminocephalosporanic acid (7-ACA) and cephalosporin C. Moreover, feruloylamino acids and L-lysine derivatives of ferulic acid derivatives were synthesized in an aqueous buffer using the enzyme. PMID- 16244441 TI - Glucoamylase originating from Schwanniomyces occidentalis is a typical alpha glucosidase. AB - A starch-hydrolyzing enzyme from Schwanniomyces occidentalis has been reported to be a novel glucoamylase, but there is no conclusive proof that it is glucoamylase. An enzyme having the hydrolytic activity toward soluble starch was purified from a strain of S. occidentalis. The enzyme showed high catalytic efficiency (k(cat)/K(m)) for maltooligosaccharides, compared with that for soluble starch. The product anomer was alpha-glucose, differing from glucoamylase as a beta-glucose producing enzyme. These findings are striking characteristics of alpha-glucosidase. The DNA encoding the enzyme was cloned and sequenced. The primary structure deduced from the nucleotide sequence was highly similar to mold, plant, and mammalian alpha-glucosidases of alpha-glucosidase family II and other glucoside hydrolase family 31 enzymes, and the two regions involved in the catalytic reaction of alpha-glucosidases were conserved. These were no similarities to the so-called glucoamylases. It was concluded that the enzyme and also S. occidentalis glucoamylase, had been already reported, were typical alpha glucosidases, and not glucoamylase. PMID- 16244444 TI - Tetrapyrrole metabolism is involved in lesion formation, cell death, in the Arabidopsis lesion initiation 1 mutant. AB - The Arabidopsis lesion initiation 1 (len1) mutant develops lesions on leaves without pathogen attack. The len1 plants display lesion formation as they grow under short-day conditions (SD), but not under long-day conditions (LD). This study was conducted to examine how lesion formation, viz., cell death, in len1 plants occurs under SD. I present genetic and physiological data to show that tetrapyrrole metobolism is necessary for lesion formation in len1 plants. Lesion formation was suppressed in the len1lin2 double mutant under SD. lesion initiation 2 (lin2) is another lesion mimic mutant with a defect in tetrapyrrole biosynthesis. Suppression of lesion formation in len1 plants was also observed when they were crossed with the mutants that had defects in other steps in tetrapyrrole metabolism. Suppression was correlated with reduced chlorophyll (Chl) levels in the double mutants. Furthermore, I found that dark-to-light transition caused a bleached phenotype in len1 plants, as in the case of antisense ACD1 (acd, accelerated cell death) plants. ACD1 encodes pheophorbide a oxygenase (PaO), which is involved in Chl catabolism in Arabidopsis. These results suggest that tetrapyrrole metabolism, especially Chl breakdown, might be involved in lesion formation in len1 plants. PMID- 16244443 TI - RKTS-33, an epoxycyclohexenone derivative that specifically inhibits Fas ligand dependent apoptosis in CTL-mediated cytotoxicity. AB - Cytotoxic T lymphocytes (CTLs) eliminate virus-infected cells and tumor cells by two distinct killing pathways, mediated by lytic granules containing perforin and by Fas ligand (FasL). ECH [(2R,3R,4S)-2,3-epoxy-4-hydroxy-5-hydroxymethyl-6-(1E) propenyl-cyclohex-5-en-1-one] has been shown to inhibit FasL-dependent apoptosis or the killing pathway in short-term culture. However, since ECH exhibited cell toxicity in long-term culture, we attempted the synthesis of less toxic epoxycyclohexenone derivatives. In the present study, we found that RKTS-33 [(2R,3R,4S)-2,3-epoxy-4-hydroxy-5-hydroxymethyl-cyclohex-5-en-1-one] has cell toxicity lower than ECH in long-term culture, and further investigated the inhibitory effect of RKTS-33 on CTL-mediated killing pathways. RKTS-33 did not affect cell-surface expression of FasL upon CD3 stimulation, but profoundly inhibited the FasL-dependent killing pathway mediated by CD4+ and CD8+ CTLs, indicating that RKTS-33 specifically blocks target cell apoptosis but not CTL function. By contrast, RKTS-33 did not affect the perforin-dependent killing pathway in CD8+ CTLs. These results indicate that RKTS-33 is a specific inhibitor of the FasL-dependent killing pathway in CTL-mediated cytotoxicity. PMID- 16244445 TI - Structural stabilities of recombinant scombridae fish myoglobins. AB - An expression system of recombinant myoglobins (Mb) of 3 scombridae fish species was constructed. The stability of these Mbs was compared with native Mbs purified from slow skeletal muscle. The addition of hemin during the cultivation of an Escherichia coli strain harboring a pGEX-2T expression vector was found to be necessary to prevent recombinant Mb from degrading and to attain its proper folding. The stabilities of recombinant Mbs were generally lower than those of native Mbs, partly due to the absence of post-translational modification. The alpha-Helical content of bullet tuna recombinant Mb at 10 degrees C was the lowest (29.0%) among the recombinant Mbs examined (the values for bluefin tuna and bigeye tuna Mbs being 34.8 and 35.5%, respectively). On the other hand, the stabilities of recombinant Mbs of bluefin tuna and bigeye tuna against denaturants (urea and guanidine hydrochloride) were found to be similar, whereas bullet tuna recombinant Mb exhibited the lowest stability among these Mbs. The pattern of temperature-dependent decrease in the alpha-helical content supported these results. PMID- 16244446 TI - Enzymatic synthesis of cytidine 5'-monophospho-N-acetylneuraminic acid. AB - We have established an efficient method for enzymatic production of cytidine 5' monophospho-N-acetylneuraminic acid (CMP-NeuAc) from inexpensive materials, N acetylglucosamine (GlcNAc) and cytidine 5'-monophosphate (CMP). The Haemophilus influenzae nanE gene encoding GlcNAc 6-phosphate (GlcNAc 6-P) 2-epimerase and the Campylobacter jejuni neuB1 gene encoding N-acetylneuraminic acid (NeuAc) synthetase, both of whose products are involved in NeuAc biosynthesis, were cloned and co-expressed in Escherichia coli cells. We examined the synthesis of NeuAc from GlcNAc via GlcNAc 6-P, N-acetylmannosamine (ManNAc) 6-P, and ManNAc by the use of E. coli cells producing GlcNAc 6-P 2-epimerase and NeuAc synthetase, in expectation of biological functions of E. coli such as the supply of phosphoenolpyruvate (PEP), which is an essential substrate for NeuAc synthetase, GlcNAc phospholylation by the PEP-dependent phosphotransferase system, and dephospholylation of ManNAc 6-P. Eleven mM NeuAc was synthesized from 50 mM GlcNAc by recombinant E. coli cells with the addition of glucose as an energy source. Next we attempted to synthesize CMP-NeuAc from GlcNAc and CMP using yeast cells, recombinant E. coli cells, and H. influenzae CMP-NeuAc synthetase, and succeeded in efficient production of CMP-NeuAc due to a sufficient supply of PEP and efficient conversion of CMP to cytidine 5'-triphosphate by yeast cells. PMID- 16244447 TI - Pungent qualities of sanshool-related compounds evaluated by a sensory test and activation of rat TRPV1. AB - The detection threshold and taste characteristics of sanshools were examined by sensory evaluation, after isolating four sanshools (alpha-, beta-, gamma-, and delta-), and two hydroxy sanshools (alpha- and beta-) from the pericarp of Japanese pepper. The Scoville unit (SU) values of the four sanshools were in the range of 80,000-110,000, while those of hydroxy sanshools were 3-5 fold lower than corresponding sanshools. The pungent qualities of each sanshool were different. Burning and tingling were predominantly perceived and lasted for the longest time with alpha-sanshool. Burning and fresh for gamma-sanshool, and tingling and numbing for hydroxy alpha-sanshool were perceived. Tests on the activation of rat TRPV1 were also performed. All of them were weak agonists. Among them, gamma-sanshool was the most potent agonist, although its EC50 value of 5.3 microM was 230 fold higher than that of capsaicin. These results indicate that it would be difficult to explain the pungent quality of each sanshool simply in terms of TRPV1 activation. PMID- 16244448 TI - Quantitative analysis of sanshool compounds in Japanese pepper (Xanthoxylum piperitum DC.) and their pungent characteristics. AB - The distributions of each sanshool in the Japanese pepper plant grown in various regions and the change in composition of sanshools during maturation of the fruit were investigated. The degree of pungency, defined as the amount of a sanshool/the threshold value, was calculated, and the pungent qualities of the products were evaluated and compared. The degree of pungency and amount of a sanshool showed a positive correlation. In young leaves and flowers, the degree of pungency was less than that in the fruits, the main compound being alpha sanshool, while the two hydroxy sanshools were detected only in trace amounts. The main compound in fruits was hydroxy alpha-sanshool, whose threshold value was higher than that of alpha-sanshool. It is concluded that the pungency of Japanese pepper should be evaluated not only by the threshold values, but also by the pungent qualities, the composition of sanshools, and the usage of each product of Japanese pepper. PMID- 16244449 TI - AtFLA11, a fasciclin-like arabinogalactan-protein, specifically localized in sclerenchyma cells. AB - An arabinogalactan-protein macroarray of all 48 Arabidopsis arabinogalactan protein genes was prepared as a handy detection system for arabinogalactan protein gene expression. The major transcript in inflorescence stems was identified as AtFLA11. AtFLA11 is categorized as a fasciclin-like arabinogalactan protein that possesses a fasciclin domain with a cell adhesion function in animal cells. AtFLA11 was specifically expressed in the sclerenchyma cells of inflorescence stems and siliques, which are characterized by their thick secondary cell walls, and was confirmed by immunostaining at the protein level. The fluctuation of AtFLA11 transcripts during the maturation process of sclerenchyma cells suggests its role in the formation of the secondary cell wall. PMID- 16244450 TI - Reduction in dietary calcium/phosphorus ratio reduces bone mass and strength in ovariectomized rats enhancing bone turnover. AB - To clarify the effects of the dietary calcium (Ca)/phosphorus (P) ratio on bone mineralization under the condition of estrogen deficiency, Wistar strain female rats were ovariectomized (OVX) at 12 weeks old. At 16 weeks old, the rats were divided into three dietary groups fed varying levels of P containing 0.5% Ca: 0.25% P, Ca/P = 2; 0.5% P, Ca/P = 1; and 1.0% P, Ca/P = 0.5 respectively. This study indicates that the reduction of the dietary Ca/P ratio impairs trabecular bone turnover accompanying the acceleration of bone formation in OVX rats. PMID- 16244451 TI - Inhibitory effect of polyphenol-enriched apple extracts on mast cell degranulation in vitro targeting the binding between IgE and FcepsilonRI. AB - Extracts from immature fruit of the apple (Rosaceae, Malus sp.), which contain procyanidins (polymers of catechins) as the major ingredients, are known to inhibit histamine release from mast cells. We analyzed in this study the mechanism for the anti-allergic activity of two polyphenol-enriched apple extracts. These extracts, termed "crude apple polyphenol (CAP)" and "apple condensed tannin (ACT)", reduced the degranulation of mast cells caused by cross linking of the high-affinity receptor for IgE (FcepsilonRI) with IgE and the antigen in a dose-dependent manner. Furthermore, western blotting revealed that phosphorylation of the intracellular signal-transduction molecules caused by cross-linking of FcepsilonRI was markedly decreased by the addition of CAP or ACT. We then analyzed the effects of CAP and ACT on the binding of the IgE antibody to FcepsilonRI on mast cells, which is the first key step in the allergic reaction mediated by mast cells, and found that this binding was markedly inhibited by both CAP and ACT. These results indicate that the inhibition of binding between FcepsilonRI and IgE by either CAP or ACT was the probable cause of the suppression of mast cell activation. This is the first report demonstrating the molecular mechanism for the anti-allergic effect of procyanidin-enriched extracts from apples. PMID- 16244452 TI - Increased expression of hypothalamic NADPH-diaphorase neurons in mice with iron supplement. AB - Iron deficiency is known as the most important nutritional problem in the world. The loss of appetite is a common characteristic of iron deficiency. Iron containing heme is required as a cofactor for nitric oxide synthase (NOS) which produces nitric oxide (NO). NOS in the central nervous system has been suggested to regulate food intake. Hence, we examined the expression of hypothalamic NOS at various levels of dietary iron. ICR mice (n = 30) were randomly divided into three groups based on the level of dietary iron and fed experimental diets for 4 weeks: the normal-iron diet group (7 mg/kg diet, n = 10), the low-iron diet group (21 mg/kg diet, n = 10) and the high-iron diet group (42 mg/kg diet, n = 10). Expression of NOS in the paraventricular nucleus (PVN) and lateral hypothalamic area (LHA) of hypothalamus was examined by histochemistry for nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-diaphorase). The high-iron diet mice showed significantly higher staining intensity of NADPH-diaphorase-positive neurons in the PVN and LHA than the normal- and low-iron diet mice. PMID- 16244453 TI - Dissimilarity in Fos and Jun immunoreactivity in hypothalamic regions between obese and lean Zucker rats. AB - The obese Zucker rat, whose genotype is transmitted in an autosomal recessive fashion, is an animal model widely used in the field of obesity. The expression of the nuclear transcription factors c-Fos and c-Jun in the paraventricular nucleus (PVN) and arcuate nucleus (ARC) of the hypothalamus of obese Zucker rats was studied using immunohistochemical methods. PVN and ARC in the hypothalamus are known as centers for the control of food intake. It was observed that the numbers of c-Fos-positive and c-Jun-positive neurons in these regions decreased in obese rats compared to lean rats, and that difference was more evident in the ARC than in the PVN which has to do with the regulation of body weight. The reduction in expression in the ARC of obese rats was greater for c-Jun than for c Fos. These results suggest a possible difference in Fos immunoreactivity in hypothalamic resistance to circulating satiety factors in genetically obese Zucker rats. PMID- 16244454 TI - Electrolyzed hydrogen-saturated water for drinking use elicits an antioxidative effect: a feeding test with rats. AB - A new type of electrolyzed hydrogen-saturated (EHS) water was produced using a water-electrolyzing device equipped with a special cation exchanger. Use of the EHS water for drinking in a feeding test with rats elicited an antioxidative effect. After intraperitoneal injection of 2,2-azobis-amidinopropane dihydrochloride, urinary secretion of 8-hydroxydeoxyguanosine and hepatic formation of peroxidized lipid were significantly lessened in rats which had received the EHS water for one week. These results suggest the possibility that this drinking water shows an effect in reduction of oxidative stress in the body. PMID- 16244455 TI - Effect of an adzuki bean extract on hepatic anti-oxidant enzyme mRNAs in D galactosamine-treated rats. AB - In acute hepatic injury tests, an adzuki bean extract decreased D-galactosamine (GalN)-induced alterations in the serum alanine aminotransferase and aspartate aminotranferase activities to about 37% and 25%, respectively, although there were no significant differences in these activities between the GalN-treated group with the adzuki bean extract and the GalN-treated group without the adzuki bean extract. Furthermore, the hepatic glutathione peroxidase, glutathione reductase, and Mn- and Cu,Zn-superoxide dismutase mRNA levels in the GalN-treated group with the adzuki bean extract were higher than those in the control group and GalN-treated group without the adzuki bean extract. PMID- 16244456 TI - Substrate shape preference of Escherichia coli ribonuclease P ribozyme and holo enzyme using bottom-half part-shifting variants of pre-tRNA. AB - We showed previously that the bacterial ribonuclease P (RNase P) ribozyme has substrate shape preference depending on the concentrations of catalytically important magnesium ions. The ribozyme discriminates a canonical cloverleaf precursor tRNA from a hairpin RNA with a CCA-tag sequence at low concentrations of magnesium ions. By detailed analysis of the shape preference using the bottom half part-shifting variants of a tRNA precursor, we showed that the RNAs in a T shape structure can be substrates for the ribozyme reactions even at low concentrations of magnesium ions, and that the RNA in a natural L-shape is the best substrate for both the ribozyme and the holo enzyme. The results also showed that the position of the bottom-half part did not affect the cleavage site selection of a substrate by the enzyme. Our results are the first kinetic evidence to show the importance of the bottom-half part of tRNA molecule, and our result also showed that the holo enzyme can discriminate substrate shape as well as the ribozyme at low concentrations of metal ions. PMID- 16244457 TI - Paenibacillus macerans possesses two types of 16S rDNA copies in a genome with a length difference of twelve base pairs. AB - Two Paenibacillus macerans strains, JCM 2500T and MCRI 12, exhibited two types of 16S rDNA copies in their genomes, accompanied by a length difference of 12 bp at positions 203 to 214 (Escherichia coli numbering). The long-type sequences were newly identified for P. macerans 16S rDNA, and the copy numbers were different between the two strains. Both types of 16S rRNA were expressed in each strain, and it was predicted that the polymorphism at this position is located in helix H10, based on a comparison with the E. coli 16S rRNA secondary structure model. PMID- 16244458 TI - Identifying 6,7,4'-trihydroxyisoflavone as a potent tyrosinase inhibitor. AB - A known biotransformed compound, 6,7,4'-trihydroxyisoflavone, was identified as a potent tyrosinase inhibitor. It inhibited mushroom tyrosinase with an IC50 value of 9.2 microM, which is six times the anti-tyrosinase activity of kojic acid (IC50 = 54.4 microM). The inhibition kinetics, analyzed by Lineweaver-Burk plots, indicated 6,7,4'-trihydroxyisoflavone to be a competitive inhibitor of tyrosinase when L-tyrosine was used as a substrate. Its biosynthesis precursors and analogs, including glycitein, daidzein, and genistein, showed little anti-tyrosinase activity. The results suggest that hydroxyl groups at the C-6 and C-7 positions of the isoflavone skeleton might play an important role in the expression of tyrosinase inhibitory activity. PMID- 16244459 TI - Crystalline structure of N-(S)-2-heptyl (1R,2R)-2-(2,3 anthracenedicarboximido)cyclohexanecarboxamide that differs from its preferred conformation in the solvent used for crystallization. AB - The crystalline structure of N-(S)-2-heptyl (1R,2R)-2-(2,3 anthracenedicarboximido)cyclohexamide (1), which was crystallized from methanol, was determined by an X-ray analysis and had a different conformation from its preferred one in CD3OD by a 1H-NMR analysis. Inter- and intra-molecular CH-pi interaction in a crystal plays a very important role in crystal packing. The preferred conformation of the amide derivative in a solution allows us to exploit (1R,2R)-2-(2,3-anthracenedicarboximido)cyclohexanecarbonyl chloride as a conversion reagent to determine the absolute configuration of chiral amines by 1H NMR. PMID- 16244460 TI - Cryptic polyadenylation of transcripts of an RNA virus gene introduced into tobacco plants. AB - We constructed an expression vector for the coat protein (CP) gene and the 3' untranslated region (3' UTR) of RNA virus (sweet potato feathery mottle virus severe strain (SPFMV-S)) lacking a foreign terminator. Out of seven transgenic tobacco plants, expression of the transgene was observed in six plants. RT-PCR analysis revealed that the transcripts had a poly(A) tail, and in most of them, polyadenylation occurred on the 5' side of the 3' UTR. These results suggest that the viral sequence contains a cryptic polyadenylation signal that permits 3'-end processing of the transcripts. PMID- 16244461 TI - An isozyme of earthworm serine proteases acts on hydrolysis of triacylglycerol. AB - An enzyme catalyzing the hydrolysis of triacylglycerol was purified from an earthworm. The N-terminal amino acid sequence and the catalytic function of the purified enzyme were identical to those of Isozyme C, an isozyme of the earthworm serine proteases. No other lipase proteins were found in the earthworm cells. The isozyme might act on the hydrolysis of triacylglycerol as well as the protein decomposition. PMID- 16244462 TI - Promoting protein, a silkworm hemolymph protein promoting in vitro replication of nucleopolyhedrovirus, binds to beta-glucans. AB - A protein which bound to 125I-labeled peptidoglycan (PGN) was isolated from hemolymph of silkworm larvae. The N-terminal amino acid sequence and the molecular weight of the protein were in accord with those described for Promoting Protein (PP) from the silkworm. The binding of the protein to [125I]PGN was competitively inhibited by various beta-glucans. The binding kinetics of PGN and chitin to the protein were analyzed in a biosensor. PMID- 16244463 TI - Uncoupling of longevity and paraquat resistance in mutants of the nematode Caenorhabditis elegans. AB - To analyze the relationship between resistance to oxidative stress and longevity, we isolated three novel paraquat-resistant mutants, mev-5, mev-6, and mev-7, from the nematode Caenorhabditis elegans. They all showed the Dyf (defective in dye filling) phenotype, but not always resistance to heat or UV. Life-span extension was observed only in the mev-5 mutant at 26 degrees C. These results indicate that longevity is uncoupled with the phenotype of paraquat resistance. PMID- 16244464 TI - Tamm-Horsfall protein acts as a general host-defense factor against bacterial cystitis. AB - AIMS: Tamm-Horsfall protein (THP) is urine's most abundant protein, but its biological function has remained elusive. Recently, THP-deficient (THP(-/-)) mice were shown to have difficulty clearing Escherichia coli from the urinary bladder. It has remained unclear if interaction between THP and E. coli is specific for E. coli or if THP has a versatile ability to clear a variety of bacteria from the bladder, and act as a broad host-defense mechanism against urinary tract infection (UTI). In this study, we examined the role of THP as a protective factor against UTI caused by bacteria other than E. coli, namely Klebsiella pneumoniae and Staphylococcus saprophyticus by determining if the THP(-/-) mouse has difficulty clearing these bacteria from its bladder. METHODS: THP gene knockout mice were generated by the technique of homologous recombination. K. pneumoniae and S. saprophyticus were introduced transurethrally, in separate experiments, into the bladders of the THP(-/-) and genetically similar wild-type (THP(+/+)) mice. Urine was collected at periodic intervals and cultured to quantitate the degree of bacteriuria. Bladders were surgically removed and examined histomorphometrically to determine the intensity of inflammation. RESULTS: Results showed that both with K. pneumoniae and with S. saprophyticus, the THP(-/-) mice had more severe bacteriuria in comparison with THP(+/+) mice. The inflammatory changes in the bladder were also markedly more intense in THP(-/ ) mice with each of the bacterial species. CONCLUSIONS: These findings support the hypothesis that THP helps eliminate K. pneumoniae and S. saprophyticus from the urinary tract and acts as a general host-defense factor against UTI. PMID- 16244465 TI - The comparative morphology of the cerebellum in caprimulgiform birds: evolutionary and functional implications. AB - Interspecific variation in the structure of the avian cerebellum is poorly understood. We present the first comparison of cerebellar morphology within the avian order Caprimulgiformes. Using a range of qualitative descriptions and quantitative measurements of cerebellar morphology we compared caprimulgiform birds with hummingbirds and swifts (Apodiformes) and owls (Strigiformes), two groups that are putative sister taxa to the Caprimulgiformes. Our results demonstrate that the owlet-nightjars (Aegothelidae), nightjars (Caprimulgidae) and potoos (Nyctibiidae) are more similar to apodiforms than they are to other taxa. All of these species have a reduced anterior lobe characterized by particularly small folia II and III and a relatively large posterior lobe. The frogmouths (Podargidae) possess a markedly different cerebellum that is more similar to that of owls than any of the caprimulgiform or apodiform birds. The monotypic oilbird (Steatornis caripensis, Steatornithidae) possesses a cerebellum with some nightjar-like features and some owl-like features, but overall it too resembles an owl more than a nightjar. This cerebellar diversity within the order Caprimulgiformes has significant implications for understanding the evolutionary relationships within the order, how the avian cerebellum has evolved and whether interspecific differences in cerebellar morphology reflect behavior. PMID- 16244466 TI - Chemoarchitecture of the monotreme olfactory bulb. AB - The cyto- and chemoarchitecture of the olfactory bulb of two monotremes (shortbeaked echidna and platypus) was studied to determine if there are any chemoarchitectural differences from therian mammals. Nissl staining in conjunction with enzyme reactivity for NADPH diaphorase, and immunoreactivity for calcium binding proteins (parvalbumin, calbindin and calretinin), neuropeptide Y, tyrosine hydroxylase and non-phosphorylated neurofilament protein (SMI-32 antibody) were applied to the echidna. Material from platypus bulb was Nissl stained, immunoreacted for calretinin, or stained for NADPH diaphorase. In contrast to eutherians, no immunoreactivity for either the SMI-32 antibody or calretinin was found in the mitral or dispersed tufted cells of the monotremes and very few parvalbumin or calbindin immunoreactive neurons were found in the bulb of the echidna. On the other hand, immunoreactivity for tyrosine hydroxylase in the echidna was similar in distribution to that seen in therians, and periglomerular and granule cells showed similar patterns of calretinin immunoreactivity to eutherians. Multipolar neuropeptide Y immunoreactive neurons were confined to the deep granule cell layer and underlying white matter of the echidna bulb and NADPH diaphorase reactivity was found in occasional granule cells, fusiform and multipolar cells of the inner plexiform and granule cell layers, as well as underlying white matter. Unlike eutherians, no NPY immunoreactive or NADPH diaphorase reactive neurons were seen in the glomerular layer. The bulb of the echidna was comparable in volume to prosimians of similar body weight, and its constituent layers were highly folded. In conclusion, the monotreme olfactory bulb does not show any significant chemoarchitectural dissimilarities from eutheria, despite differences in mitral/tufted cell distribution. PMID- 16244467 TI - Cyto- and chemoarchitecture of the monotreme olfactory tubercle. AB - This study was undertaken to determine whether the olfactory tubercles of two monotremes (platypus and echidna) showed cyto- or chemoarchitectural differences from the tubercles of therian mammals. Nissl staining was applied in conjunction with enzyme reactivity for NADPH diaphorase and acetylcholinesterase, and immunoreactivity for calcium binding proteins (parvalbumin, calbindin and calretinin) and tyrosine hydroxylase (echidna only). Golgi impregnations of the tubercle were also available for the echidna. The olfactory tubercle is a poorly laminated structure in the echidna, despite the pronounced development of other components of the echidna olfactory system, and the dense cell layer of the olfactory tubercle was found to be discontinuous and irregular. Granule cell clusters (islands of Calleja) were present, but were small, poorly defined and did not show the intense NADPH diaphorase activity seen in marsupial and placental mammals. A putative small island of Calleja magna was seen in only one echidna out of four. In Golgi impregnations of the echidna olfactory tubercle, the most abundant neuron type was a medium-sized densely spined neuron similar to that seen in the olfactory tubercle of some therians. Large spine-poor neurons were also seen in the polymorphic layer. In the platypus, the olfactory tubercle was very small but showed more pronounced lamination than the echidna, although no granule cell clusters were seen. In both monotremes, the development of the olfactory tubercle was poor relative to other components of the olfactory system (bulb and piriform cortex). The small olfactory tubercle region in the platypus is consistent with poor olfaction in that aquatic mammal, but the tubercle in the echidna is more like that of a microsmatic mammal than other placentals occupying a similar niche (e.g., insectivores). PMID- 16244468 TI - Relative medial and dorsal cortex volume in relation to sex differences in spatial ecology of a snake population. AB - In non-avian reptiles the medial and dorsal cortices are putative homologues of the hippocampal formation in mammals and birds. Studies on mammals and birds commonly report neuro-ecological correlations between hippocampal volume and aspects of spatial ecology. We examined the relationship between putative homologous cortical volumes and spatial use in a population of the squamate reptile, Agkistrodon piscivorus, that exhibits sex differences in spatial use. Do male A. piscivorus that inhabit larger home ranges than females also have larger putative hippocampal volumes? Male and female brains were sectioned and digitized to quantify regional cortical volumes. Although sex differences in dorsal cortex volume were not observed, males had a significantly larger medial cortex relative to telencephalon volume. Similar to studies on mammals and birds, relative hippocampal or medial cortex volume was positively correlated with patterns of spatial use. We demonstrate volumetric sex differences within a reptilian putative hippocampal homologue. PMID- 16244469 TI - Elevation of cytokine concentrations in asphyxiated neonates. AB - BACKGROUND: Various cytokines are reportedly associated with many neonatal diseases. Asphyxia is considered to result in ischemia-reperfusion injuries and induces abnormal inflammatory responses involving excessive cytokine production. OBJECTIVES: To evaluate alteration in sera levels of various cytokines/chemokines in case of perinatal asphyxia at birth. METHODS: In order to determine the concentrations of various cytokines/chemokines in sera, we used a highly sensitive fluorescence microsphere method. We measured the concentration of 8 types of cytokines/chemokines in sera obtained from 17 cases of asphyxia, 10 normal neonates, and 6 healthy adults. RESULTS: The concentrations of IL-6, IL-8, and IL-10 in the sera of asphyxiated neonates were higher than those in the normal neonates. Irrespective of the presence or absence of asphyxia, sera concentrations of IL-2, IL-4, IFN-gamma, and TNF-alpha were higher in the neonates than those in the adults. The concentration of IFN-gamma in the asphyxiated neonates was lower than that in the normal neonates. Sera levels of IL-10 were higher in the asphyxiated cases than those in the normal neonates. The sera levels of IL-6, IL-8, and IL-10 in asphyxiated neonates with either a poor outcome or death were higher than those without poor outcomes. CONCLUSIONS: The concentrations of various types of cytokines/chemokines were different in neonatal sera and some of them increased drastically during asphyxia. The concentration of an anti-inflammatory cytokine IL-10 was elevated in asphyxiated neonates immediately after birth, thereby suggesting that IL-10 might be associated with neuroprotective functions. PMID- 16244470 TI - Impact of a genetic polymorphism of the interleukin-1 receptor antagonist on technique survival in peritoneal dialysis patients. AB - BACKGROUND/AIMS: There is a clear association between one allele of the interleukin-1 receptor-antagonist gene (IL-1RN) and inflammatory diseases in which IL-1 is implicated. We evaluated patient survival and technique survival of peritoneal dialysis (PD) patients, while analyzing independent risk factors, in a PD program. We also tested the association between IL-1RN polymorphism, patient survival and technique survival. METHODS: We retrospectively evaluated 129 Japanese CAPD patients undergoing initial treatment in eight centers in Japan. Using PCR, IL-1RN genotype and allele frequencies were determined, and clinical and biochemical variables were recorded at the start of PD. The relation of patient survival or technique survival with IL-1RN polymorphism and those variables was analyzed with a multivariate Cox's proportional-hazard model. RESULTS: The frequencies of IL-1RN*1/IL-1RN*1 and IL-1RN*1/IL-1RN*2 genotypes were 84.5 and 15.5%, respectively. Median patient survival was 37.0 months, and overall patient survival was 92.8 and 87.9% at 2 and 5 years, respectively. Age, cardiovascular disease and serum albumin were found to be independent predictors of patient survival. Median technique survival was 32 months. PD failure occurred in 37 patients, with technique survival rates of 92.0 and 72.7% at 2 and 5 years, respectively. Serum albumin, peritonitis and the presence of the IL-1RN*2 genotype were found to be independent predictors of technique survival. CONCLUSION: Serum albumin was the strongest predictive factor for mortality and technique failure in PD. Technique failure was also affected by IL-1RN polymorphism in this patient population. PMID- 16244472 TI - Introduction. Genes and patents. PMID- 16244471 TI - Metabolic acidosis in peritoneal dialysis patients: the role of residual renal function. AB - BACKGROUND: Metabolic acidosis (MA) is common in chronic renal insufficiency (CRI) patients, and its pattern changes as renal function deteriorates. Although the prevalence of acidosis in peritoneal dialysis has been reported to be rather high, the causes of it have not been well studied. The present study was performed to examine the prevalence of metabolic acidosis in our continuous ambulatory peritoneal dialysis (CAPD) patients and its possible causes. METHODS: In this cross-sectional study, we analyzed data from patients who received maintenance CAPD in our hospital and had been on dialysis for at least one month. Patients' demographic features, medications, and intercurrent medical conditions were recorded. Data including blood biochemistry, dialysis adequacy, and nutrition were collected. A serum bicarbonate concentration of less than 23 mmol/l was defined as having acidosis. The normal value of the serum anion gap (AG) was defined as 12+/- 4 mmol/l. RESULTS: A total of 154 patients (76 males and 78 females) with age of 60.04+/- 13.92 years and the time on dialysis of 16.83+/- 21.59 months were included in this study. Sixty-six patients (43%) had a serum bicarbonate of less than 23 mmol/l, among whom 12 patients (8%) were identified as having MA with increased AG, 54 (35%) were identified as having MA with normal AG. Patients who had better residual renal function (RRF) had a significantly lower serum bicarbonate level despite their higher total KT/V(urea) as compared to those with lower RRF. In addition, patients with MA and normal AG had the highest RRF and highest total KT/V(urea). All patients with MA and increased AG had significantly lower values of dietary protein intake (DPI) as compared to their values of normalized protein nitrogen appearance (nPNA), and had higher serum urea and phosphate levels as compared with those patients without MA. CONCLUSION: Our study suggested that CAPD patients with better RRF were more susceptible to metabolic acidosis, which was characterized by normal anion gap and hyperchloremia. Thus, we speculate that renal loss of bicarbonate may to a large extent be responsible for the occurrence of MA in these patients. PMID- 16244474 TI - Genetic patent protection in the pharmaceutical and biotechnology industries. AB - Without patent protection, biomedical progress would be severely diminished. Conditions under the current patent regime are characterized by rapid advancement made possible by cooperative licensing, collaboration and partnerships between and among various entities, and the drive to bring successful products to market both in order to make profits and to further the cause of humanity. The financial advantages associated with patent-driven corporate participation are the lifeline of innovation. While granting limited periods of exclusivity under the patent system necessary to entice innovation is a calculated sacrifice, the enormous benefits of fully-disclosed pharmaceutical and genetic discoveries result in a handsome net benefit over the alternative of resource-limited research clouded by a shroud of secrecy as a substitute for patent protection. By examining characteristics of the pharmaceutical and biotechnology industries and the critical role the patent regime plays in driving investment in these areas, a clearer picture of the necessity of strong intellectual property rights in the context of genetics will emerge. PMID- 16244473 TI - What are gene patents and why are people worried about them? AB - This article examines what it means to patent a gene. Numerous ethical concerns have been raised about the effects of such patents on clinical medical practice as well as on research and development. We describe what kinds of inventions are covered by human gene patents, give several examples and summarize the small body of empirical research performed in the US examining the effects of these patents. There is little evidence that early fears about gene patenting placing substantial restraints on research and clinical medicine have come to fruition. Nonetheless, there are areas of concern, and policy makers, physicians and the public should be alert to ensure that the net social benefits of patenting human genes are maintained. PMID- 16244475 TI - Patenting and licensing of university-based genetic inventions--a view from experience at Stanford University's Office of Technology Licensing. AB - University technology transfer offices license inventions created at universities to companies looking for tools, additions to their product pipelines, improvements or start-up opportunities. This paper will cover the motivations for conducting university technology transfer, an overview of Stanford's Office of Technology Licensing and its patenting and licensing practices, models of royalty sharing at university technology transfer offices, and patenting and licensing genetic information inventions at Stanford University. The terms 'invention' and 'technology' are used interchangeably throughout the paper. PMID- 16244476 TI - Policy conflicts: gene patents and health care in Canada. AB - Several recent gene patent controversies have energized and refocused the human gene patent debate in Canada. These include the use of the Myriad test for breast cancer by the provinces, patenting of the Severe Acute Respiratory Syndrome virus and a recent Supreme Court decision rejecting the patenting of 'higher life forms'. These cases place the emerging policy conflicts between the innovation and commercialization agenda of the government and the desire to provide equitable access to health care in sharp focus. Another challenge faced by Canada is the powerful influence of the United States in policy decisions. Although these issues have raised awareness about the possibility of reforming the patent system, Parliament has yet to consider any of the suggested reforms of the Canadian patent system and there are no formal proposals pending. PMID- 16244477 TI - Balancing innovation and access to healthcare through the patent system--an Australian perspective. AB - This article examines the enforcement of gene and other research tool patents in Australia. An empirical analysis of patenting practices in the Australian medical biotechnology industry showed heightened concern about the impact of patents on research and diagnostic testing, but provided little evidence to support these concerns at that time. Since then, the Australian company Genetic Technologies Ltd. has been enforcing its patents for non-coding DNA sequences. The governments of Australia are encouraging the biotechnology industry to better protect and enforce intellectual property rights, but recognize these needs to be balanced against access to healthcare. The article discusses proposals made by the Australian Law Reform Commission to adjust the balance, both by tightening the requirements for obtaining patents and by introducing various options to assist providers of diagnostic services and others in using patented inventions, but at the same time maintaining the incentive to innovate. PMID- 16244478 TI - The patent is political: the consequences of patenting the BRCA genes in Britain. AB - OBJECTIVES: The paper explores the attempt by an American biotechnology company, Myriad Genetics, to use its patent rights over the BRCA genes to transfer its technology of genetic testing for breast and ovarian cancer to Britain. It also investigates the responses of British scientists, health care professionals and patient advocates to this attempted technology transfer. METHODS: This paper is based on approximately 100 in-depth interviews, document analysis and ethnographic observation conducted in the United States and Britain from 1998 to 2001. RESULTS: The BRCA gene patents inspired political resistance and mobilized opposition to the patenting of genes in general. They also provided an opportunity for the British to assert their national identity as they argued that a British BRCA testing service needed to be available within the context of the National Health Service to all citizens equally. CONCLUSIONS: Patents are not only legal documents and technical descriptions, but political tools as well. As they are increasingly deemed vital to economic globalization, patents have become mobilizing tools for anti-globalization activists and non-governmental organizations from less developed countries, and for asserting local and national identities. PMID- 16244479 TI - Visual search in patients with subcortical vascular dementia: short fixations but long reaction times. AB - Visual search is a cognitive function of high ecological relevance. It involves rapid alternations between allocating and shifting attention. In patients with Alzheimer's disease, the duration of fixations during visual search increases already in the early stage of the illness. Subcortical vascular dementia (SVD), a newly defined subgroup of vascular dementia, has not yet been examined in this respect. SVD affects patients with a history of lacunar infarctions and/or transient ischemic attacks, focal neurological signs and evidence of subcortical white matter lesions as well as lacunes in the deep grey matter. Here, we report our findings from tracking eye movements during a visual search task with different array sizes in 9 patients with SVD and compare the number and duration of eye fixations they made with the values obtained in 9 healthy elderly control subjects. While patients with SVD were significantly slower in the tasks with longer center to target distances (mean reaction time), the number and duration of fixations they made did not differ from those in controls. Impairment of visual search in patients with SVD seems to be an effect of general cognitive slowing in more demanding arrays of visual search rather than a specific deficit in parameters of eye fixation. PMID- 16244480 TI - Memory for performed actions in dementia of Alzheimer type: further evidence for a global semantic memory deficit. AB - Findings in patients with dementia of the Alzheimer type (AD patients) concerning the enactment effect, i.e. reaching a better memory performance after encoding actions by performing them in contrast to encoding them by reading, are ambiguous. In order to get more insight into memory deterioration in patients suffering from AD, an experimental study was run. Patients with mild-to-moderate AD (n = 23) and healthy elderly control persons (n = 20) took part. They had to encode actions verbally as well as enactively. Memory performance was tested by using recognition judgments instead of the predominating free recall. The AD patients showed no enactment effect and a rate of false alarm judgments that was more than twice as high as found in healthy elderly controls. This enactive encoding failure may be indicative of memory deficits underlying the conceptual system of action representations. PMID- 16244481 TI - Olanzapine versus haloperidol in the treatment of agitation in elderly patients with dementia: results of a randomized controlled double-blind trial. AB - The goal of this study was to compare the efficacy and safety of olanzapine versus haloperidol in the treatment of agitation and aggression in patients with dementia. The subjects were 58 out-patients with dementia and agitation. After baseline assessments and, if necessary, a period of wash-out of a previous antipsychotic drug, they were randomly assigned to 5 weeks of double-blind treatment with either olanzapine or haloperidol. The first 2 weeks were used for dose titration. Subsequently, the patients received a fixed dose of either olanzapine (average dose 4.71 mg) or haloperidol (average dose 1.75 mg) from day 14 to day 35. Both olanzapine and haloperidol decreased agitation significantly (decrease in Cohen-Mansfield Agitation Inventory scores), but there was no significant difference between the two drugs. The two drugs had comparable effects on all secondary outcome measures. They were well tolerated and had a similar side-effect pattern. Our study could not demonstrate the superiority of olanzapine, compared to haloperidol, for the treatment of agitation in patients with dementia. PMID- 16244482 TI - Measurement of thirteen biological markers in CSF of patients with Alzheimer's disease and other dementias. AB - Cerebrospinal fluid (CSF) biological markers may be of valuable help in the diagnosis of dementia. The aim of the present study was to evaluate CSF levels of 13 potential biomarkers in patients with Alzheimer's disease (AD), frontotemporal lobe dementia, alcohol dementia, major depression and control patients without any neuropsychiatric disease. The study was performed using beta-amyloid 1-42 (Abeta42), total tau and phosphorylated tau-181 (P-tau181) as core markers. The ratio P-tau181/Abeta42 could significantly distinguish AD patients from all other diagnostic subgroups. CSF levels of 5 growth factors (HGF, GDNF, VEGF, BDNF, FGF 2) and 3 cytokines/chemokines (TNF-alpha, TGF-beta1, MIP-1alpha) did not significantly differentiate between the studied groups. However, depending on the degree of neurodegeneration (as expressed by the ratio P-tau181/Abeta42), patients with AD displayed significantly increased CSF levels of nerve growth factor (NGF) as compared to healthy controls. CSF levels of monocyte chemoattractant protein 1 (MCP-1) were found to be significantly increased with age in all groups but did not distinguish AD patients from healthy controls. The results confirmed the suitability of the ratio P-tau181/Abeta42 for the diagnosis of AD, while CSF levels of NGF and MCP-1 are less specific and reliable for AD. It is suggested that the increase in NGF depends on the extent of neurodegeneration of the AD type and the increase in MCP-1 on age. PMID- 16244483 TI - Atrophic gastritis is associated with increased sucrose permeability related to chronic inflammation. AB - BACKGROUND: Different theories have been presented to explain how atrophic gastritis may lead to gastric cancer development. One contributing factor could be impaired function of the gastric mucosal barrier. The aim of this study was to investigate if there are changes in gastric mucosal permeability to sucrose in atrophic gastritis. METHODS: The study comprised 22 patients with atrophic gastritis and 21 normal controls. Gastritis was classified according to the Sydney system from endoscopic biopsies of the gastric corpus and antrum. All subjects were exposed to oral sucrose load (100 g), and the fraction of sucrose excreted in urine was measured by gas chromatography-mass spectrometry. RESULTS: The fraction of sucrose excreted in urine after oral load was significantly increased in atrophic gastritis compared with controls (median 0.08 vs. 0.04%; p = 0.003). Sucrose excretion was positively related to the degree of chronic inflammation (median fraction excreted: mild inflammation 0.06%, moderate inflammation 0.08%, severe inflammation 0.18%; p = 0.04) rather than to the degree of atrophy in the gastric mucosa. Occurrence of intestinal metaplasia was also associated with significantly higher sucrose excretion. However, in multivariate analysis, including intestinal metaplasia, only the degree of inflammation was positively related to sucrose excretion. CONCLUSION: Atrophic gastritis is associated with increased sucrose permeability, suggesting paracellular leakage of the gastric mucosa. This leakage seems to be related to the degree of inflammation rather than the degree of atrophy. The findings may have implications for the diseases and complications associated with atrophic gastritis. PMID- 16244484 TI - A note on aphasia in bilingual patients: Pitres' and Ribot's laws. AB - Johann Gesner in 1770 probably provided the first description of dissociation in reading ability in different languages in a bilingual patient, who after brain damage was able to read Latin but not German. Clinical studies have since shown that bilingual 'aphasics' do not necessarily manifest the same language disorders with the same degree of severity in both languages. Superficially, different case findings indicate instances of shared and divergent representation of components of language in the bilingual brain. This paper considers a selection of many empirical studies, which have failed to reconcile the parallel recovery of language in many reported bilingual aphasiacs and the differential recovery in others. It reviews Pitres' rule (recovery of the most used acquired language) and Ribot's law (recovery of the native language) that are important concepts during recovery and rehabilitation of bilingual aphasiacs. PMID- 16244485 TI - Virginia Apgar (1909-1974): neurological evaluation of the newborn infant. PMID- 16244486 TI - Theodore Herpin: neglected contributions in the understanding of epilepsy. AB - In contrast to earlier notions of epilepsy, Theodore Herpin considered that when symptoms started in the periphery or the viscera their origin was in the brain, a view later lauded by Hughlings Jackson. His respected book Des acces incomplets did not attempt to deal with disordered physiology but was designed to help physicians to recognise and treat epilepsy early in its course. He described juvenile myoclonic epilepsy and vividly and accurately portrayed many other clinical phenomena of epilepsy. PMID- 16244487 TI - Intravenous TPA for very old stroke patients. AB - BACKGROUND: Although thrombolysis in patients with advanced age is considered more risky, some may benefit from TPA treatment. We studied safety and recanalization/recovery in patients older than 80 years treated with TPA and compared them with younger stroke patients. METHODS: We studied patients treated with intravenous TPA and divided them into younger (<80 years) and older (> or =80 years) groups for comparison. Diagnostic transcranial Doppler was completed before bolus, and patients were consequently monitored for up to 2 h when feasible. Clinical data included NIH Stroke Scale score, symptomatic intracranial hemorrhage (ICH) and discharge disposition. RESULTS: We studied 127 younger (mean 63 years, range 31-79) and 56 older patients (mean 84 years, range 80-93). Median baseline NIH Stroke Scale score was higher in the older group (18 vs. 14 points, NS). Occlusion locations, onset to needle time (median 130 vs. 120 min) as well as improvement at 24 h (median 5 vs. 4 points) were similar in both groups. Transcranial Doppler monitoring showed similar partial or complete recanalization rates (66 vs. 66%), onset to recanalization time (median 160 vs. 158 min) and reocclusion rates (26 vs. 25%). Symptomatic and fatal ICH was not higher in the older group (7.1 and 3.5% vs. 6.3 and 3.9%, NS). There was higher mortality among older patients (20 vs. 11%, NS). At discharge, 23% of older patients went home, 41% underwent rehabilitation and 16% were transferred to skilled nursing facilities, compared with 31, 43 and 15% respectively, in the younger group. CONCLUSION: After intravenous TPA treatment, patients over 80 years of age have similar recanalization, short-term improvement and symptomatic ICH rates compared with younger patients. However, older patients tend to have higher in-hospital mortality. PMID- 16244488 TI - Treatment of endometriosis with transvaginal ultrasound-guided drainage under GnRH analogues and recombinant interleukin-2 left in the cysts. AB - BACKGROUND: To analyze the therapeutic results of one dose of 3 million IU of recombinant interleukin-2 (rIL-2) left intracyst (group I) versus two doses with a 1-month interval (group II) after transvaginal ultrasound (US)-guided drainage of endometriomas under the effect of GnRH analogues. METHODS: Prospective and randomized clinical trial (helped by a random number table) at a University Hospital. Twenty-four consecutive patients with endometriomas initially sent to us for laparotomy and conservative surgery for endometriosis were included. INTERVENTIONS: Treatment with GnRH analogues every 28 days, 3 doses. Under their effect, one or two transvaginal US-guided punctures were performed in order to aspirate the endometriomas, and 3 million IU of rIL-2 were left in the aspirated cysts each time. MAIN OUTCOME MEASURES: CLINICAL RESULTS: two menstruations after GnRH analogues. Other secondary outcome measures were: the time until recurrence of cysts, symptoms and CA-125 >35 U/ml, and the need for further medical or surgical treatment. RESULTS: They were moderate or good in >50% of cases with one drainage and one dose of 3 million IU of rIL-2 intracyst, but were better with a second drainage and two doses of rIL-2 (25 vs. 58.3% 'good results'). There were fewer recurrences and the interval before recurrence was longer after two doses but differences were not significant. Six patients from group I (50%) and 3 from group II (25%) needed laparotomy and conservative surgery at 17.5 +/- 8.7 months (total time of follow-up = 33 +/- 8.8 months). CONCLUSIONS: Treatment of endometriomas with transvaginal US-guided drainage and rIL-2 left in the cysts under endometrial suppressive therapy with GnRH analogues has beneficial effects, improving clinical manifestations and avoiding some surgical therapies. The use of a higher dose of rIL-2 does not produce better results, whereas drainage + rIL-2 twice does. PMID- 16244489 TI - A rapidly growing paraurethral myoma with profuse bleeding from a mucosal vessel: report of a case. AB - Paraurethral leiomyoma is rare. This is the first reported case of a woman with a 6 x 7 x 5 cm urethral leiomyoma with profuse vaginal bleeding. The site of bleeding was identified as the anterior vaginal mucosal vein, which we subsequently electrocauterized. Enucleation of the mass was performed smoothly with a Foley catheter to avoid damage to the urethra. Profuse bleeding from a paraurethral myoma is possibly due to increased vascularity and the prolapsed nature of the tumor in this area. PMID- 16244490 TI - Association of endometriosis risk and genetic polymorphisms involving sex steroid biosynthesis and their receptors: a meta-analysis. AB - Endometriosis is a sex steroids-dependent disease. It has been postulated that certain genetic polymorphisms involved in sex steroids biosynthesis and metabolisms may be associated with increased risk of developing endometriosis. Despite a deluge of reports of positive associations of endometriosis with numerous polymorphisms involving sex steroids production and metabolism, the results are often conflicting. We performed a meta-analysis of 12 association studies on 5 genes (CYP17, CYP19, AR, PR and ER). We found that many reported positive findings were not supported by the data due to faulty analysis. There have been no functional data that support a putative relationship of these genetic polymorphisms with endometriosis. A handful of positive findings so far have not been independently replicated, and should be viewed as preliminary. In addition, these findings should be counterbalanced by legitimate concerns of multiple comparisons, small prior probability of association with a particular polymorphism, proper selection of controls, and lack of replication (at least until now). In future association studies, it may be productive to put more thought to study design, execution, and data analysis. PMID- 16244491 TI - MRI evaluation of the uterine structure after myomectomy. AB - Myomectomy is a good indication for women with uterine leiomyoma who desire to preserve their child-bearing potential. However, there are still no reports about how long it takes the uterus to reach a stable state after myomectomy. We evaluated the changes in uterine structure during the recovery process after myomectomy by MR images. MR images were used to analyze the time-dependent changes in the length of the uterine cavity, the volume of the uterus, recovery of the junctional zone, prevalence of modification of the endometrium, and uterine structure in the region of the enucleated myoma. The cavity length and the volume of the uterus, and the myometrium were stabilized at six weeks after the myomectomy. With regard to the endometrium, 12 weeks were required for it to achieve a stable state after myomectomy. However, even at 12 weeks postoperatively, 14.2% of the cases showed an unusual view near the uterine incision on MR images. We concluded that the recovery process is complete at 12 weeks after the operation if there are no clear findings of hematoma or edema formation in the myometrium on MR images. PMID- 16244492 TI - Two successful pregnancies following eight miscarriages in a patient with antithrombin deficiency. AB - Inherited thrombophilias are associated with an increased risk of maternal thromboembolism and certain adverse pregnancy outcomes, including second- and third-trimester fetal loss, placental abruption, severe intrauterine growth restriction, and early-onset, severe preeclampsia. Pregnant patients with severe thrombophilias, especially antithrombinopathies are at very high risk for both thromboembolism and adverse pregnancy outcomes. A case of a patient with antithrombin deficiency is reported, who had two successful pregnancies after eight miscarriages. Our case shows that a combined treatment with antithrombin substitution and a prophylactic, body-weight-adjusted dose of low-molecular weight heparin may be successful in preventing pregnancy loss and thromboembolism in antithrombin deficiency during pregnancy, although other complications, such as preeclampsia and intrauterine growth restriction cannot always be prevented. PMID- 16244493 TI - A novel autosomal recessive non-syndromic deafness locus, DFNB66, maps to chromosome 6p21.2-22.3 in a large Tunisian consanguineous family. AB - Hereditary non-syndromic deafness is extremely heterogeneous. Autosomal recessive forms account for approximately 80% of genetic cases. Autosomal recessive non syndromic sensorineural deafness segregating in a large consanguineous Tunisian family was mapped to chromosome 6p21.2-22.3. A maximum lod score of 5.36 at theta=0 was obtained for the polymorphic microsatellite marker IR2/IR4. Haplotype analysis defined a 16.5-Mb critical region between microsatellite markers D6S1602 and D6S1665. The screening of 3 candidate genes, COL11A2, BAK1 and TMHS, did not reveal any disease causing mutation, suggesting that this is a novel deafness locus, which has been named DFNB66. A search in the Human Cochlear EST Library for ESTs located in this critical interval allowed us to identify several candidates. Further investigations on these candidates are needed in order to identify the deafness-causing gene in this Tunisian family. PMID- 16244494 TI - Leptin directly controls secretory activity of human ovarian granulosa cells: possible inter-relationship with the IGF/IGFBP system. AB - AIMS: The aim of our in vitro studies was to understand the role of leptin and the insulin-like growth factor I/insulin-like growth factor protein (IGF/IGFBP) system in controlling human ovarian function. METHODS: We studied the action of leptin (0, 1, 10, or 100 ng/ml) and immunoneutralization of IGF-I using specific antiserum (0.1%) on the release of progesterone (P), estradiol (E), oxytocin (OT), IGF-I, IGFBP-3, and prostaglandins F (PGF) by these cells using radioimmunoassay/immunoradiometric assay. RESULTS: It was found that leptin stimulated the secretion of OT, IGFBP-3, and PGF. It suppressed the secretion of E and IGF-I, but not P, into the medium. The addition of antiserum against IGF-I decreased IGF-I output, increased P, OT, IGFBP-3, and PGF secretion, and had no effect on E release. Immunoneutralization of IGF-I also prevented or reversed the effects of leptin on P, E, IGF-I, IGFBP-3, PGF, but not on OT. CONCLUSIONS: These observations (1) demonstrate that leptin directly controls the secretory activity of human ovarian cells, (2) confirm the involvement of IGF-I in the regulation of ovarian cells, and (3) suggest an inter-relationship between leptin and the IGF/IGFBP system in the control of these functions and the involvement of IGF/IGFBP system in mediating leptin action on the ovary. PMID- 16244495 TI - Ultrasound-microbubble-mediated NF-kappaB decoy transfection attenuates neointimal formation after arterial injury in mice. AB - OBJECTIVE: Decoy transfection is a significant methodology for suppressing gene activation. The decoy can be synthesized easily; however, the major obstacle is the difficulty involved in effective in vivo delivery. METHODS AND RESULTS: We used a fluorescein-labeled decoy to investigate the ultrasound-microbubble mediated in vivo delivery in normal and injured mouse arteries. We showed that this approach could deliver the decoy into target tissues. In addition, we performed in vivo NF-kappaB decoy transfection into murine injured arteries using the ultrasound-microbubble method. Murine femoral arteries were injured using flexible wires to establish arterial injury. Pathologically, neointima/media areas in the NF-kappaB decoy transfection using ultrasound-microbubble group showed less than those in the control groups. Immunohistochemistry revealed that enhanced expression of inflammatory factors was observed in nontreated injured arteries, while the NF-kappaB decoy suppressed the expression. CONCLUSION: We revealed that ultrasound-microbubble delivery of the decoy is effective for transfection into target organs. We also indicated that NF-kappaB decoy transfection using this method has potential for the suppression of neointimal formation. Ultrasound-mediated transfection of the decoy can be beneficial for the clinical treatment of restenosis after coronary intervention and other cardiovascular diseases. PMID- 16244496 TI - Effect of chronic viral hepatitis on graft survival in Saudi renal transplant patients. AB - BACKGROUND: In Saudi Arabia the prevalence of hepatitis C among hemodialysis patients is very high ranging from 60 to 80%. A large number of these dialysis patients go for renal transplant, resulting into a higher prevalence of hepatitis C virus (HCV) infection in renal transplant patients. Yet no current systematic report is available on the influence of hepatitis C status on patient and graft survival. The present study was therefore undertaken to address this objective. METHODS: Retrospective analysis of data of 448 renal transplantation subjects was undertaken. The mean follow-up period was 5.85 +/- 2.7 (median 5.3) years. The factors associated with renal graft survival were reviewed and these include: age, sex, and type of donor, immunosuppressive medication, episodes of infection, blood pressure, serum creatinine, and status of hepatitis. The primary end-points were renal graft function and patient survival. Logistic regression, COX regression analysis, and Kaplan-Meier survival estimates were used to evaluate the influence of hepatitis C on the above parameters. RESULTS: Among 448 recipients of first kidney transplant patients, 286 (63.8%) were positive for HCV infection. In the HCV-positive group, 204 (71.32%) were males. Kaplan-Meier survival analysis showed a significantly better graft survival for HCV-negative patients than HCV-positive patients (p < 0.001; log-rank test). Logistic regression analysis and COX regression analysis have shown different grades of graft dysfunction were present in HCV-positive patients after adjustment for covariates: age, sex, blood pressure, type of donor, and immunosuppressive medication; the presence of HCV was a major predictor of bad outcome and significantly influenced graft survival (odds ratio = 4.37; 95% Cl = 1.81-4.77). Significant deterioration of liver function was noted in HCV-positive patients at the last follow-up, taking ALT as a marker (ALT level 80.6 +/- 5.8 U/l at the last follow-up versus 49.5 +/- 32 U/l at baseline p < or = 0.0001). Sixteen patients had a chronic active course and 1 patient developed biopsy-proven liver cirrhosis and portal hypertension. A serious and significantly greater incidence of fatal chest infections was seen in HCV-positive patients. Although mortality was greater in HCV-positive versus HCV-negative patients (20 vs. 7), the difference did not attain statistical significance (p = 0.23) and none of the patients died as a result of hepatic failure. CONCLUSION: The presence of HCV infection greatly influenced graft survival in renal transplant patients and a higher proportion of infected patients had renal and hepatic dysfunction. A significant increase in fatal chest infections was noted in HCV-positive patients. Overall mortality was higher in HCV-positive patients, but it was not statistically significant. All measures should be taken to prevent HCV transmission in the dialysis population. Renal transplant recipients with HCV infection need close monitoring for both graft and liver function. PMID- 16244498 TI - Acute and chronic regulation of the renal Na/H+ exchanger NHE3 in rats with STZ induced diabetes mellitus. AB - BACKGROUND: Early stages of diabetic nephropathy are characterized by alterations of glomerular filtration, increased tubular sodium and water reabsorption, and systemic volume expansion, which may be a major cause for the development of hypertension. As a significant fraction of renal salt and water transport is mediated by the proximal tubular Na+/H+ exchanger NHE3, we investigated its regulation in rats with STZ-induced diabetes mellitus. METHODS: Male Sprague Dawley rats were injected +/- streptozotocin (STZ, 60 mg/kg), and sacrificed after 2, 7 or 14 days. Renal cortical BBM vesicles were prepared to measure Na+/H+ exchange (NHE) activity and NHE3 protein abundance. Cortical NHE3 mRNA was extracted to perform Northern blot analysis. Pharmacological inhibitors were used in vivo and in vitro in order to identify isoform specificity conferring changes in NHE activity mediated by the diabetic milieu. RESULTS: Compared to control rats, STZ rats were clearly hyperglycemic at all time points studied. NHE activity was significantly increased by 40 and 37% in diabetic rats after 7 and 14 days, respectively, but not after 2 days. The increase in Na+/H+ exchange activity was not inhibited by HOE-642 (3 microM). Administration of exogenous insulin to diabetic rats resulted in lower blood sugars, but not NHE activity. Moreover, serum glucose concentration did not correlate with NHE activity in any subgroup nor in all animals analyzed together. However, in STZ rats supplemented with exogenous insulin NHE activity was positively correlated with serum insulin concentrations (r = 0.86, p < 0.01). In vivo, the increase in NHE activity induced by STZ could be completely inhibited when rats were fed 6 ppm of HOE-642 with the diet over 14 days. The changes in Na+/H+ exchange activity were not paralleled by changes in NHE3 protein or mRNA abundance in diabetic rats at any of the time points investigated. CONCLUSIONS: These results suggest that proximal tubular Na/H exchange activity is modified in the early stage of diabetes mellitus. PMID- 16244497 TI - Expression analysis of hypothalamic and pituitary components of the growth hormone axis in fasted and streptozotocin-treated neuropeptide Y (NPY)-intact (NPY+/+) and NPY-knockout (NPY-/-) mice. AB - In the fasted and the streptozotocin (STZ)-induced diabetic male rat, hypothalamic growth hormone (GH)-releasing hormone (GHRH) mRNA levels, and pulsatile GH release are decreased. These changes are believed to be due to a rise in hypothalamic neuropeptide Y (NPY) that inhibits GHRH expression. To directly test if NPY is required for metabolic regulation of hypothalamic neuropeptides important in GH secretion, NPY, GHRH and somatostatin (SRIH) mRNA levels were determined in fasted (48 h) and STZ-treated wild-type (NPY(+/+)) and NPY-knockout (NPY(-/-)) mice by ribonuclease protection assay. In addition, pituitary receptor mRNA levels for GHRH (GHRH-R), ghrelin (GHS-R) and SRIH (sst2) were assessed by RT-PCR. Under fed conditions the GH axis of NPY(+/+) and NPY(-/ ) did not differ. In the NPY(+/+) mouse, fasting resulted in a 23% weight loss and >250% increase in NPY mRNA accompanied by a significant reduction in both GHRH and SRIH mRNA. These changes were associated with increases in pituitary expression of GHRH-R and GHS-R and a concomitant suppression of sst2. In the NPY( /-) mouse, fasting also resulted in a 23% weight loss and comparable changes in GHRH-R and sst2, but failed to alter GHRH, SRIH and GHS-R mRNA levels. Fasting resulted in an overall increase in circulating GH, which reached significance in the fasted NPY(-/-) mouse. Induction of diabetes in NPY(+/+) mice, using a single, high-dose, STZ injection (150 mg/kg), resulted in modest weight loss (5%), and a 158% increase NPY expression which was associated with reciprocal changes in pituitary GHS-R and sst2 expression, similar to that observed in the fasted state, but no change in hypothalamic GHRH or SRIF expression was observed. Induction of diabetes in NPY(+/+) and NPY(-/-) mice, using a multiple, low-dose, STZ paradigm (5 consecutive daily injections of 40 mg/kg), did not alter body weight, hypothalamic neuropeptide expression or pituitary receptor expression, with the exception that sst2 mRNA levels were suppressed and GH levels did rise in the NPY(-/-) mouse. These observations demonstrate that NPY is not required for basal regulation of the GH axis, but is required for fasting-induced suppression of GHRH and SRIH expression, as well as fasting-induced augmentation of pituitary GHS-R mRNA. In contrast to the rat, fasting clearly did not suppress circulating GH levels in mice, but resulted in an overall rise in mean GH levels, similar to that observed in other mammalian species. The fact that many of the fasting-induced changes in the GH axis were observed in the high-dose STZ-treated mice, but were not observed in the multiple, low-dose paradigm, suggests STZ mediated modulation of GH axis function is dependent on the severity of the catabolic state and not hyperglycemia. PMID- 16244499 TI - Phosphorylation of neuronal and endothelial nitric oxide synthase in the kidney with high and low salt diets. AB - BACKGROUND: Renal nitric oxide (NO) synthesis increases with increasing salt intake, however, the mechanisms underlying this are poorly understood. We hypothesized that activating or inhibitory phosphorylation of neuronal and endothelial nitric oxide synthase (nNOS, eNOS) regulates renal NO production in response to altered dietary salt. METHODS: Sprague-Dawley rats were fed low, normal or high salt diets for 12 h or 2 weeks, and kidney NOS phosphorylation was analyzed by Western blot using phosphopeptide antibodies against the sites nNOS Ser(1412), nNOS-Ser(847), eNOS-Ser(1176) and eNOS-Thr(494). RESULTS: At 12 h, total nNOS increased 1.4-fold (p < 0.01) in the high salt group and decreased by 26% (p < 0.05) in the low salt group. Changes in expression of phospho-nNOS at 12 h were accounted for by the changes in total nNOS. No change in total or phospho eNOS was seen at 12 h. At 2 weeks, in the low salt group expression of total nNOS increased 1.8-fold (p < 0.05) whereas expression of nNOS phosphorylated at the inhibitory site Ser(847) increased 4.3-fold (p < 0.01). Total eNOS was increased 3-fold in the low salt group (p < 0.01), with parallel increases in eNOS phosphorylated at both activating and inhibitory sites (p < 0.05). In the 2-week high salt group no changes in NOS expression or phosphorylation were seen, despite the observed increased excretion of urinary NO metabolites. CONCLUSION: In summary, changes in phospho-nNOS and phospho-eNOS expression occurred in parallel with changes in total expression, thus, the overall activating and inhibitory effects of nNOS and eNOS phosphorylation at the sites studied were not changed by altered dietary salt. PMID- 16244500 TI - Evidence for a tetrahydrobiopterin deficit in schizophrenia. AB - Tetrahydrobiopterin (BH(4)) is a vital cofactor maintaining availability of the amine neurotransmitters [dopamine (DA), noradrenaline (NA), and serotonin (5 HT)], regulating the synthesis of nitric oxide (NO) by nitric oxide synthase (NOS), and stimulating and modulating the glutamatergic system (directly and indirectly). These BH(4) properties and their potential relevance to schizophrenia led us to investigate the hypothesis of a study group (healthy controls, n=37; schizophrenics, n=154) effect on fasting plasma total biopterin levels (a measure of BH(4)). Study analysis showed a highly significant deficit of total biopterins for the schizophrenic sample after partialling out the effects of potential confounds of gender, age, ethnicity, neuroleptic use history and dose of current use, 24-hour dietary phenylalanine/protein ratio (a dietary variable relevant to BH(4) synthesis), and plasma phenylalanine (which stimulates BH(4) synthesis). A mean decrement of 34% in plasma total biopterins for schizophrenics from control values supports clinical relevance for the finding. In a subsample (21 controls and 23 schizophrenics), sequence analysis was done of the GTP cyclohydrolase I feedback regulatory gene and no mutations were found in the coding region of the gene. A deficiency of BH(4) could lead to hypofunction of the systems of DA, NA, 5-HT, NOS/NO, and glutamate, all of which have been independently implicated in schizophrenia psychopathology. Further, evidence has been accumulating which implicates the critical interdependence of these neurotransmitter systems in schizophrenia; this concept, along with the present study finding of a biopterin deficit, suggests that further study of the BH(4) system in schizophrenia is warranted and desirable. PMID- 16244501 TI - Interleukin-6 genetic polymorphism and Chinese major depression. AB - BACKGROUND: It has been reported that serum concentrations of interleukin-6 (IL 6), a pro-inflammatory cytokine, were significantly higher in major depressive disorder (MDD). OBJECTIVES: The aim of the present study was to examine whether the IL-6 genetic variant confers susceptibility to MDD. METHODS: We genotyped a functional polymorphism in the promoter region (position -634) of the IL-6 gene in a sample of 257 MDD patients and 105 normal controls. RESULTS: No significant differences were demonstrated for the genotype or allele frequency of the IL-6 polymorphism when comparing the MDD and control groups. CONCLUSIONS: Our findings suggested that the investigated IL-6 polymorphism does not affect MDD susceptibility. We propose that other genetic variations of IL-6, relating either to MDD or to the clinical manifestations of MDD, may need further investigation. PMID- 16244502 TI - Cortisol and heart rate measures during casino gambling in relation to impulsivity. AB - Problematic gambling behavior is thought to be influenced by neurobiological as well as environmental factors. In this study, we investigated the relationship among impulsivity, gambling behavior, the cardiovascular system and the hypothalamic-pituitary-adrenal axis activity in blackjack gamblers. Twenty-nine males were continuously monitored before, during and after a 90-min blackjack session in a casino wagering their own money and during a control condition where subjects played cards for accumulation of points. Heart rate and cortisol levels significantly increased with the onset of gambling and remained elevated throughout the test session compared to the control condition. After median split of impulsivity scores, high impulsivity subjects revealed significantly higher heart rate levels compared to the low impulsivity subgroup. Correlation analyses revealed a positive relationship between impulsivity scores and severity of pathological gambling. Impulsivity may be one important factor mediating gambling behavior and its accompanying autonomic response. PMID- 16244504 TI - Anaemia of cancer: impact on patient fatigue and long-term outcome. AB - Anaemia is the most common haematological abnormality encountered by cancer patients. A large European survey of cancer patients (n = 15,367) reported that 67% had anaemia at some point during the survey, and that over 60% of these patients did not receive any treatment for their anaemia. Two other surveys (the FATIGUE surveys) showed that over 75% of cancer patients experienced fatigue at least monthly, with over 30% reporting this symptom on a daily basis. Significantly, patients regarded fatigue as having a greater negative impact on their daily lives than many other cancer- or treatment-related complications, with important emotional and mental consequences including lack of self motivation, sadness, frustration, and mental exhaustion. Indeed, fatigue was considered so debilitating, 12% of patients felt their quality of life (QoL) was so reduced that they did not wish to continue living. Anaemia is also recognised as an independent predictor of poor prognosis in cancer patients. A systematic review evaluating survival showed a 65% overall increase in the risk of mortality in cancer patients with anaemia. Increasing physicians' awareness of the importance of effectively treating anaemia in cancer patients therefore has the potential to improve prognosis as well as QoL. PMID- 16244505 TI - Anaemia management strategies: optimising treatment using epoetin beta (NeoRecormon). AB - Anaemia has a detrimental impact on quality of life and it is important that this condition is recognised and treated in patients with cancer. Epoetin beta is an effective and well-tolerated treatment of anaemia in patients with a wide range of solid and haematological malignancies. A study in patients with lymphoid malignancies confirms that epoetin beta is equally effective at the same overall weekly dose (30,000 IU weekly) when given once-weekly or three-times weekly. This once-weekly regimen has also proved effective in patients with solid tumours. Once-weekly treatment is more convenient for the patient, potentially improving compliance and is associated with reduced hospital administration costs. The majority of patients with cancer will respond to epoetin therapy with an increase in haemoglobin levels. However, it is of value to identify those patients who are likely to respond, so that cost-effectiveness can be improved. Despite much research into potential predictive factors, follow-up studies are required and clinical judgement remains key to managing the anaemia of cancer. In addition, studies suggest that intravenous iron supplementation can improve response to epoetin therapy in patients with functional iron deficiency. Epoetin beta offers an effective, safe and convenient therapy for the management of anaemia in patients with cancer. Ongoing studies are expected to lead to a greater understanding of the optimal use of epoetins in cancer-related anaemia. PMID- 16244506 TI - New guidelines on anaemia management in patients with cancer: How do these affect clinical practice? AB - Anaemia is a condition that frequently occurs in patients with cancer, but a recent survey has cast doubt over whether it is being appropriately treated. These findings are disappointing considering the wealth of data showing the effectiveness of epoetin therapy in patients with cancer. Recently, evidence based guidelines have been published that aim to provide physicians with the latest information required for optimal management of anaemia in patients with cancer. By increasing physician awareness of the appropriate therapy for anaemia management, more patients may receive the benefits of epoetin therapy. PMID- 16244507 TI - Beyond anaemia management: evolving role of erythropoietin therapy in neurological disorders, multiple myeloma and tumour hypoxia models. AB - Recombinant human erythropoietin (epoetin) has become the standard of care in the treatment of anaemia resulting from cancer and its treatment, and chronic kidney disease. The discovery that erythropoietin and its receptor are located in regions outside the erythropoietic system has led to interest in the potential role of epoetin in other tissues, such as the central nervous system. Animal studies have shown that systemically applied epoetin can cross the blood-brain barrier, where it reduces tissue injury associated with stroke, blunt trauma and experimental autoimmune encephalomyelitis. Pilot studies in humans have shown that epoetin treatment given within 8 h of stroke reduces infarct size and results in a significantly better outcome when compared with placebo treatment. Studies also suggest that epoetin has the potential to improve cognitive impairment associated with adjuvant chemotherapy in patients with cancer. Anaemia is a major factor causing tumour hypoxia, a condition that can promote changes within neoplastic cells that further tumour survival and malignant progression and also reduces the effectiveness of several anticancer therapies including radiotherapy and oxygen-dependent cytotoxic agents. Use of epoetin to prevent or correct anaemia has the potential to reduce tumour hypoxia and improve treatment outcome. Several therapeutic studies in anaemic animals with experimental tumours have shown a beneficial effect of epoetin on delaying tumour growth. Furthermore, clinical observations in patients with multiple myeloma and animal studies have suggested that epoetin has an antimyeloma effect, mediated via the immune system through activation of CD8+ T cells. Therefore, the role of epoetin may go well beyond that of increasing haemoglobin levels in anaemic patients, although additional studies are required to confirm these promising results. PMID- 16244508 TI - By whom and how is the quality of research data collection assured and checked? PMID- 16244509 TI - Technological innovations in clinical assessment and psychotherapy. AB - In this paper the application of computer technology and the use of the Internet in mental health care are critically reviewed. A number of on-line screening devices have been developed for anxiety disorders, mood disorders, and substance abuse disorders, with great potential for clinical practice. On line assessment is generally equivalent to clinical assessment. A number of studies have shown that self-help treatment programmes on stand-alone computers are as effective as routine clinical care. The Internet enhances the therapeutic possibilities of computers by offering feedback of therapists and more tailor-made treatment. A number of randomized clinical trials (RCTs) have shown that Internet-based treatment is more effective than no-treatment and as effective as face-to-face treatment. Research so far has been limited to anxiety disorders, burn-out, depression, headache, insomnia, tinnitus and obesity. Further, exposure through virtual reality has been found effective in a number of RCTs in specific phobias, but results with respect to the effects of the use of virtual reality techniques to other disorders are inconclusive. It is concluded that computer-driven assessment and treatment has many advantages and few disadvantages. A number of reasons are discussed which will preclude large-scale implementation of computer driven assessment and therapy in the near future. PMID- 16244510 TI - Psychological and quality-of-life outcomes from a comprehensive stress reduction and lifestyle program in patients with coronary artery disease: results of a randomized trial. AB - BACKGROUND: Stress reduction and comprehensive lifestyle modification programs have improved atherosclerosis and cardiac risk factors in earlier trials. Little is known about the impact of such programs on quality-of-life (QoL) and psychological outcomes. Given recent significant improvements in cardiac care, we evaluated the current benefit of stress reduction/lifestyle modification on QoL and emotional distress in patients with coronary artery disease (CAD). METHODS: 101 patients (59.4 +/- 8.6 years, 23 female) with CAD were randomized to a 1-year lifestyle/stress management program (n = 48) or written advice (n = 53). QoL and psychological outcomes were assessed with the SF-36, Beck Depression, Spielberger State/Trait Anxiety, Spielberger State/Trait Anger and Perceived Stress Inventories. Group repeated-measures analyses of variance were performed for all measures. RESULTS: Adherence to the program was excellent (daily relaxation practice 39 +/- 5 vs. 5 +/- 8 min, respectively; p < 0.001). Both groups improved comparably in most dimensions of QoL, and significantly greater improvements for the lifestyle group were found for physical function and physical sum score (p = 0.046 and p = 0.045). Depression, anxiety, anger and perceived stress were reduced similarly in both groups. However, intervention x gender interaction effects revealed greater benefits among women in the lifestyle intervention vs. advice group for depression and anger (p = 0.025 and p = 0.040), but no effects for men. CONCLUSIONS: A comprehensive lifestyle modification and stress management program did not improve psychological outcomes in medically stable CAD patients. The program did appear to confer psychological benefits for women but not men. Further trials should investigate gender-related differences in coronary patient responses to behavioral interventions. PMID- 16244511 TI - Comparison of changes in physical self-concept, global self-esteem, depression and anxiety following two different psychomotor therapy programs in nonpsychotic psychiatric inpatients. AB - BACKGROUND: The first objective of this study was to compare the changes in physical self-concept, global self-esteem, depression and anxiety after participation in one of two 16-week psychomotor therapy programs for nonpsychotic psychiatric inpatients. The second objective was to study the relationship between changes in these variables. METHODS: One hundred and ninety-nine inpatients were randomly assigned to either a personalized psychomotor fitness program, consisting of aerobic exercise and weight training, or a general program of psychomotor therapy, consisting of different forms of physical exercises and relaxation training. Physical self-concept was evaluated using the Dutch version of the Physical Self-Perception Profile at baseline, after 8 weeks, and after completion of the 16-week interventions. At the same time points, additional variables of global self-esteem, depression and anxiety were assessed by means of the Rosenberg Self-Esteem Inventory, the Beck Depression Inventory and the Trait Anxiety Inventory, respectively. RESULTS: After 16 weeks, both groups showed significant improvements in all outcome measures (p values ranged from 0.01 to < 0.0001), with no between-group differences. In both groups, the improvement in physical self-concept was correlated with increased global self-esteem and decreased depression and anxiety levels (p < 0.01). CONCLUSIONS: The results suggest that both psychomotor therapy programs are equally effective in enhancing physical self-concept. The relationship between improvements in physical self concept and enhancements in global self-esteem, depression and anxiety supports the potential role of the physical self-concept in the recovery process of depressed and anxious psychiatric inpatients. PMID- 16244512 TI - Antidepressant response and well-being in pre-, peri- and postmenopausal women with major depressive disorder treated with fluoxetine. AB - BACKGROUND: We assessed the impact of menopausal status on treatment response and well-being in a cohort of outpatient women with major depressive disorder (DSM III-R criteria), who received treatment with fluoxetine (20 mg/day for 8 weeks). METHODS: Menopausal status was defined based on age, presence of menstrual irregularity or amenorrhea and vasomotor symptoms. Remission and response of depression were defined as a 17-item Hamilton Depression Rating Scale (HAM-D-17) score or=50%, respectively. Well-being was assessed by self-rating with the Symptom Questionnaire. Remitters were followed up for 28 additional weeks. RESULTS: No differences in rates of response and remission as well as in levels of well-being were observed among pre- (n = 121), peri- (n = 28) and postmenopausal (n = 35) women at the endpoint of the acute phase, even after adjustment for baseline depression severity. Residual symptoms, however, were significantly more common in postmenopausal women, except for the continuation phase endpoint. Differences in residual symptoms during the acute phase subsided after adjustment for baseline depression severity. CONCLUSIONS: Overall, menopausal status did not significantly affect the response to fluoxetine treatment and the degree of posttreatment well-being among major depressive disorder patients. PMID- 16244513 TI - In-patient treatment of mothers with depressive disorders - does joint admission of their children have a negative effect on the results of treatment? AB - BACKGROUND: There is disagreement whether in-patient psychotherapeutic treatment results for women whose children are cohospitalised with them would be better if the mothers were hospitalised without children. The purpose of this study was to examine this question in the case of mothers with depressive symptoms. METHODS: The treatment results for 43 female in-patients (21 with and 22 without accompanying children) in a prospective, randomised, controlled study were compared. The period of observation was 6 weeks. The Beck Depression Inventory (BDI) and the Changes in Experience and Behaviour Questionnaire (VEV) were used for outcome measures. RESULTS: At 6 weeks, according to the intent-to-treat principle, neither the VEV (p = 0.58) nor the BDI (p = 0.57) yielded significant differences between mothers whose children were admitted jointly and mothers whose children were not. CONCLUSION: The treatment results for patients with depressive symptoms whoare accompanied by their children for the duration of their in-patient psychotherapy treatment are just as good as those for mothers whose children are not jointly admitted. Additionally, the data have further implications that lend themselves to discussion regarding support for this type of facility. PMID- 16244514 TI - Stability of alexithymia and its relationships with the 'big five' factors, temperament, character, and attachment style. AB - BACKGROUND: Controversy still exists concerning the stability of the alexithymia construct. Also, although alexithymia has been found to be related in a theoretically meaningful way to other personality constructs such as the 'Big Five' factors, few studies have investigated its relationship with influential constructs such as temperament and character, and attachment security. METHODS: Two hundred twenty-one undergraduate and graduate students were administered the Toronto Alexithymia Scale (TAS-20), the State-Trait Anxiety Inventory (STAI), the Zung Depression Scale (ZDS), the Temperament and Character Inventory (TCI-125), the Big Five Questionnaire (BFQ), and the Experiences in Close Relationships (ECR) questionnaire. After 1 month, 115 participants completed again the TAS-20, STAI, and ZDS. RESULTS: Alexithymia was only moderately correlated with depression and anxiety. Both the absolute and relative stability of TAS-20 total and subscale scores was high, and a negligible portion of their change over time was accounted for by changes in depression or anxiety. In separate multiple regression models including also gender, age, depression and anxiety, TAS-20 total and subscale scores were correlated with low energy/extraversion, low emotional stability, openness, low friendliness/agreeableness; harm avoidance, low self-directedness, low cooperativeness, low reward dependence; attachment related avoidance and anxiety. CONCLUSIONS: Our findings lend support for both absolute and relative stability of alexithymia, corroborate an association between alexithymia and insecure attachment, and contribute to a coherent placing of alexithymia in the broader theoretical network of personality constructs. PMID- 16244516 TI - Interferon-alpha-induced depression: when a randomized trial is not a randomized controlled trial. PMID- 16244515 TI - Coping with serious accidental injury: a one-year follow-up study. AB - BACKGROUND: The aim of this study was to analyze changes of coping strategies in severely injured accident victims over time and to compare patients with high and low posttraumatic stress disorder (PTSD) symptom levels with regard to their coping patterns and accident-related cognitions. METHODS: 106 consecutive patients with severe accidental injuries admitted to a trauma surgery intensive care unit (ICU) were assessed within 1 month after the trauma and 6 and 12 months later. Assessments included a clinical interview, the Freiburg Questionnaire of Coping with Illness, the patients' accident-related cognitions, the Clinician Administered PTSD Scale, the 90-item revised Symptom Checklist (SCL-90-R), and the Sense of Coherence Questionnaire (SOC). Patients who met the criteria for either full or subsyndromal PTSD at least once over the observation period (36 subjects; 34.0%) were assigned to a highly symptomatic group (HSG), the remainder (70 subjects; 66.0%) to a less symptomatic group. RESULTS: Overall, active problem-focused coping was predominant immediately after the accident and declined over time, with a stronger decrease in the HSG. Patients in the HSG scored higher on the SCL Global Severity Index and lower on the SOC. The patients' subjective appraisal of accident severity was higher in the HSG, whereas there was no group difference with regard to accident-related variables such as type of accident, injury severity and mild to moderate traumatic brain injury. CONCLUSIONS: Active problem-focused coping, although utilized most frequently and often regarded as protective, might be an inadequate strategy in face of acute stress following a severe accident. Clinicians should not expect their patients to cope very actively in the acute ICU phase. In the subsequent rehabilitation, active coping seems to be more adaptive. PMID- 16244517 TI - Completed suicide among subjects receiving psychotherapy. PMID- 16244518 TI - Galactorrhea with aripiprazole. PMID- 16244519 TI - Conducting multiple-site clinical trials in medical rehabilitation research. AB - This article examines the distinctive opportunities and challenges of multiple site clinical trials, as distinguished from trials conducted by a single program. Among the topics discussed are: the role of clinical trials generally in medical rehabilitation research; definitions of "clinical trials" and "multiple site"; the distinction between exploratory and capstone trials; the potential advantages and disadvantages of multiple-site trials; planning issues in terms of sampling designs, the choice of outcome measures, approaches to dealing with intervention fidelity, organizational structures, authorship, and the conduct of collateral studies; and implementation issues in terms of pilot-testing procedures, handling emerging issues, and optimizing communication. PMID- 16244520 TI - Intramuscular electrical stimulation for hemiplegic shoulder pain: a 12-month follow-up of a multiple-center, randomized clinical trial. AB - OBJECTIVE: Assess the effectiveness of intramuscular electrical stimulation in reducing hemiplegic shoulder pain at 12 mos posttreatment. DESIGN: A total of 61 chronic stroke survivors with shoulder pain and subluxation participated in this multiple-center, single-blinded, randomized clinical trial. Treatment subjects received intramuscular electrical stimulation to the supraspinatus, posterior deltoid, middle deltoid, and upper trapezius for 6 hrs/day for 6 wks. Control subjects were treated with a cuff-type sling for 6 wks. Brief Pain Inventory question 12, an 11-point numeric rating scale was administered in a blinded manner at baseline, end of treatment, and at 3, 6, and 12 mos posttreatment. Treatment success was defined as a minimum 2-point reduction in Brief Pain Inventory question 12 at all posttreatment assessments. Secondary measures included pain-related quality of life (Brief Pain Inventory question 23), subluxation, motor impairment, range of motion, spasticity, and activity limitation. RESULTS: The electrical stimulation group exhibited a significantly higher success rate than controls (63% vs. 21%, P = 0.001). Repeated-measure analysis of variance revealed significant treatment effects on posttreatment Brief Pain Inventory question 12 (F = 21.2, P < 0.001) and Brief Pain Inventory question 23 (F = 8.3, P < 0.001). Treatment effects on other secondary measures were not significant. CONCLUSIONS: Intramuscular electrical stimulation reduces hemiplegic shoulder pain, and the effect is maintained for > or =12 mos posttreatment. PMID- 16244521 TI - Corticosteroid treatment and functional improvement in Duchenne muscular dystrophy: long-term effect. AB - OBJECTIVE: To determine and compare the long-term effects of prednisone and deflazacort on the functional status of children with Duchenne muscular dystrophy. DESIGN: A total of 49 boys with Duchenne muscular dystrophy, between the age of 12 and 15 yrs, who were observed over a 7-yr period were reviewed retrospectively. Eighteen had been treated with prednisone, 12 with deflazacort, and 19 had no drug treatment. All boys treated with steroids received medication for >2 yrs before losing their ambulation. Lower and upper limb motor functions, pulmonary function, prevalence of surgery for scoliosis, and side effects were compared. RESULTS: Boys in the steroid groups were significantly more functional and performed better on all tests than boys not treated (P < 0.05). There was no significant difference between the deflazacort- and prednisone-treated groups (P > 0.05). The number of boys having scoliosis surgery in treated groups was significantly less than nontreated boys (P < 0.05). The control group's pulmonary capacity was decreasing and significantly less than both prednisone- and deflazacort-treated boys. Both deflazacort and prednisone had beneficial effect on pulmonary function and scoliosis. Cataracts, hypertension, behavioral changes, excessive weight gain, and vertebral fracture were noted as serious side effects. CONCLUSIONS: Prednisone and deflazacort have a significant beneficial effect on slowing the disease progress. Their usage in Duchenne muscular dystrophy may prolong ambulation and upper limb function with similar potency. Both steroids also improve pulmonary function, in addition to delaying the need for spinal interventions, with similar therapeutic profiles. PMID- 16244522 TI - Alternatives to endotracheal intubation for patients with neuromuscular diseases. AB - OBJECTIVE: To evaluate the usefulness of continuous noninvasive mechanical ventilation and mechanical coughing aids to avoid endotracheal intubation and tracheostomy during episodes of acute respiratory failure in patients with neuromuscular disease. DESIGN: We conducted a prospective cohort study at the respiratory medicine ward of a university hospital to study the success rate of the use of continuous noninvasive mechanical ventilation and manually and mechanically (CoughAssist) assisted coughing to avert endotracheal intubation in 24 consecutive episodes of acute respiratory failure for 17 patients with neuromuscular disease. The noninvasive mechanical ventilation and coughing aids were used to reverse decreases in oxyhemoglobin saturation and relieve respiratory distress that occurred despite oxygen therapy and appropriate medication. Noninvasive mechanical ventilation was delivered by volume ventilators (Breas PV 501) alternating nasal/oronasal and oral interfaces. RESULTS: Noninvasive management was successful in averting death and endotracheal intubation in 79.2% of the acute episodes. There were no significant differences in respiratory function between the successfully treated and unsuccessfully treated groups before the current episode. Bulbar dysfunction was the independent risk factor for failure of noninvasive treatment (P < 0.05; odds ratio, 35.99%; 95% confidence interval, 1.71-757.68). CONCLUSIONS: Intubation can be avoided for some patients with neuromuscular disease in acute respiratory failure by some combination of noninvasive mechanical ventilation and mechanically assisted coughing. Severe bulbar involvement can limit the effectiveness of noninvasive management. PMID- 16244523 TI - Effect of a lateral wedge on joint moments during gait in subjects with recurrent ankle sprain. AB - This study assessed the biomechanic effects of wearing a lateral wedge on the subtalar joint moment during gait in athletes with and without an unstable lateral ankle. A crossover design was applied whereby 50 collegiate males walked with two different wedges: a 0 degrees control wedge and a 6 degrees lateral wedge. We investigated frontal plane angles and moments at the subtalar joint and the knee joint, as well as ground reaction forces and center of pressure excursion. Moments were derived using a three-dimensional inverse dynamics model of the lower extremity. The 6 degrees lateral wedge significantly increased the subtalar joint valgus moment (P < 0.001) and reduced the knee joint varus moment (P < 0.001) when compared with no wedge. The differences between wedge conditions were associated with a laterally shifted location of the center of pressure during stance phase. However, there were diverse, sometimes reversed effects with the wedge among the athletes with an unstable lateral ankle. These results indicate that biomechanical indications and limitations of lateral wedges in unstable lateral ankles should be analyzed in more detail, possibly leading to new guidelines for the use of such foot orthoses. PMID- 16244524 TI - Effects of regular resistance training on muscle histopathology and morphometry in elderly patients with chronic kidney disease. AB - OBJECTIVE: To evaluate muscle histopathological outcomes, adaptation in muscle fiber area, and fiber type proportion after a resistance training program for patients with chronic kidney disease. DESIGN: Vastus lateralis muscle biopsies before and after the study period in seven patients and six healthy subjects performing the exercise program and in five patients in a nonexercising comparison group. RESULTS: The change in degree of histopathological abnormality did not differ between the groups after the exercise program. Muscle fiber type proportion and muscle fiber area was also the same after the observation period. CONCLUSIONS: Patients with chronic kidney disease have muscle histopathological abnormalities already in the predialysis phase. There was no indication that the exercise program had disadvantageous effects on muscle histopathology in these patients. There were no differences in muscle fiber area or in fiber type proportion after the exercise program within or between the groups. Thus, a workload of 60% of one repetition maximum was sufficient to increase muscular strength and endurance in patients with chronic kidney disease but not to increase muscle fiber area or to change muscle fiber type proportion. PMID- 16244525 TI - Effect of penile vibratory stimulation on spasticity in men with spinal cord injury. AB - OBJECTIVE: Penile vibratory stimulation is the first treatment option for anejaculation in men with spinal cord injury. It has been postulated that it also has an antispasticity effect. The purpose of this study was to determine the effect of penile vibratory stimulation on spasticity in patients with spinal cord injury. DESIGN: Ten male patients with spinal cord injury were included in the study. They were examined at the baseline and 3, 6, 24, and 48 hrs later. After their baseline examination, they performed penile vibratory stimulation. The outcome variables were spasticity (Ashworth scale), spasm frequency, spasm severity, painful spasms, plantar stimulation response, deep tendon reflexes, clonus, and effect on function. RESULTS: Ashworth grade showed a statistically significant decrease at hour 3 and hour 6 examinations (P = 0.001 and P = 0.03, respectively, with Tukey test). The patients showed a tendency toward having less frequent and less severe spasms throughout the study; however, it did not reach to a significant level. Similarly, clonus showed a nonsignificant decrease during follow-up examinations. The other variables did not change considerably. CONCLUSIONS: Penile vibratory stimulation may contribute to the relief of the spasticity in men with spinal cord injury. In treating spasticity, all the factors that increase or decrease the tone should be considered. PMID- 16244527 TI - Current status of nonsteroidal anti-inflammatory drugs in physiatry: balancing risks and benefits in pain management. AB - The gastropathy associated with the administration of nonsteroidal anti inflammatory drugs (NSAIDs) for musculoskeletal pain disorders has contributed to significant morbidity and mortality. The distinction between the cyclooxygenase 1 and 2 enzymatic properties lead to the development of selective cyclooxygenase-2 inhibitory NSAIDs with the prospect of reducing NSAID-related gastropathy while maintaining anti-inflammatory properties. Initial studies of the efficacy and safety of the selective cyclooxygenase-2 inhibitors seemed promising. Larger clinical trials were carried out to reinforce the efficacy and safety of the cyclooxygenase-2 anti-inflammatory medications (coxibs). Further analysis of these trials raised concern with regard to both the efficacy and safety of this class of drugs. The most recent clinical trials of the coxibs have demonstrated significant cardiovascular thrombogenic potential, particularly at higher doses. Clinical investigators and regulatory agencies have questioned whether these findings mitigate the efficacy of coxibs and NSAIDs in general in the prophylaxis of colonic polyps, Alzheimer's disease, and more saliently, musculoskeletal pain disorders. This article addresses the current controversy of the efficacy and safety of the coxibs and NSAIDs in general based on recent clinical trials and review by healthcare consortiums. This article also provides guidelines regarding the use of NSAIDs, including the diminishing armamentarium of the coxibs, and the alternative therapeutic options available to the physiatrist in managing musculoskeletal pain disorders. PMID- 16244526 TI - Acute inpatient rehabilitation of 55 patients after liver transplantation. AB - OBJECTIVE: Acute rehabilitation for patients who have had liver transplants is often necessary to restore functional mobility, but no studies, except for case reports, have documented the complications, outcomes, or predictors of success of an acute inpatient rehabilitation program. Our objective was to examine each of these areas related to rehabilitation after liver transplantation. DESIGN: We performed a chart review of 55 patients who received a liver transplant and were treated on our acute rehabilitation service. We examined several factors, including age, reason for transplant, length of acute hospital stay, length of acute inpatient rehabilitation stay, FIMtrade mark scores, albumin levels, medical complications, and discharge disposition. RESULTS: Patients undergoing rehabilitation after liver transplantation required multiple medical interventions, including liver biopsy, ultrasound, blood transfusions, and dialysis. Several patients (15%) were transferred to the acute care hospital for medical problems. Both a low albumin and a long length of stay in the acute care hospital correlated with a longer stay in rehabilitation and less efficient rehabilitation. Longer rehabilitation stays correlated positively with FIM gain (P < 0.001, r = 0.534). Significant improvements in FIM scores (P < 0.001) during acute inpatient rehabilitation were noted in patients who were discharged to home. We also found that age, albumin level, and length of stay in the hospital did not predict discharge disposition. CONCLUSION: Patients who have had liver transplants can achieve significant functional gains in acute rehabilitation. Rehabilitation professionals should be aware of potential complications and factors that may affect the rehabilitation of this patient population. PMID- 16244528 TI - Occult periarthrosis of the shoulder: a possible progenitor of tennis elbow. AB - Nineteen outpatients, nine women and ten men, all right-handed, ranging in age from 33 to 87 yrs, with an average age of 53.5 yrs, presented with complaints of lateral epicondylitis (i.e., tennis elbow). Each was subsequently identified as having a heretofore unrecognized loss of ipsilateral shoulder internal rotation. The authors suggest that in this scenario the occult shoulder periarthrosis is interlinked in a pathokinetic chain potentially predisposing to the presenting symptoms of tennis elbow. During a tennis serve, excessive wrist flexion may be substituted as to compensate for a restricted arc of shoulder internal rotation. The extensor carpi radialis longus and brevis cross both the elbow and wrist. Functionally, they have the capacity of reversing their osseous attachments, both at their origin and insertion, as these muscles cycle under load from a concentric/shortening to an eccentric/elongating contraction. Aging muscle, with its increased viscoelastic stiffness, may be "too slow" to accommodate this transition recoil, especially when repetitive. Instead, it is pathomechanically absorbed at the proximal elbow extensor tendon insertion. PMID- 16244529 TI - Bilateral adrenal hemorrhage: an unusual complication after bilateral total knee arthroplasty. AB - Total knee arthroplasty is a commonly performed surgical procedure usually associated with excellent improvement in mobility and function and with lessening of pain. Most cases are performed without complication. Complications, when they do occur, are most often associated with deep venous thrombosis and possible pulmonary embolism, infection, prosthetic failure or loosening, or neurovascular complications. This article presents a case report of an unusual, but serious, complication, bilateral adrenal hemorrhage, occurring in the postoperative, rehabilitative phase after what seemed to be successful bilateral knee arthroplasties in an 83-yr-old woman. PMID- 16244530 TI - Serial electrodiagnostic studies in West Nile virus-associated acute flaccid paralysis. AB - A man in his 70s presented for acute rehabilitation with severe acute flaccid asymmetric weakness in both lower limbs. Cerebrospinal fluid and serum immunoglobulin M titers were positive for West Nile virus. Electrodiagnostic studies demonstrated severe diffuse motor axonopathy consistent with an anterior myelitis. Electrodiagnostic and clinical improvements were monitored. Electrodiagnostic testing at 6 and 18 mos demonstrated continuing reinnervation; nascent voluntary motor unit action potentials were first noted proximally and, at 18 mos, distally in the left lower limb, including muscles in which motor unit potentials were not initially noted. Corresponding clinical improvements, though slow, were demonstrated even at 1(1/2) yrs after onset. Thus, motoric changes after West Nile virus-associated anterior myelitis need to be monitored over a prolonged time period to allow accurate assessment of prognosis for recovery in rehabilitation programs. PMID- 16244531 TI - Lumbar synovial cyst. PMID- 16244532 TI - Effects of a malpractice crisis on specialist supply and patient access to care. AB - OBJECTIVE: To investigate specialist physicians' practice decisions in response to liability concerns and their perceptions of the impact of the malpractice environment on patient access to care. SUMMARY BACKGROUND DATA: A perennial concern during "malpractice crises" is that liability costs will drive physicians in high-risk specialties out of practice, creating specialist shortages and access-to-care problems. METHODS: Mail survey of 824 Pennsylvania physicians in general surgery, neurosurgery, orthopedic surgery, obstetrics/gynecology, emergency medicine, and radiology eliciting information on practice decisions made in response to rising liability costs. RESULTS: Strong majorities of specialists reported increases over the last 3 years in patients' driving distances (58%) and waiting times (83%) for specialist care or surgery, waiting times for emergency department care (82%), and the number of patients forced to switch physicians (89%). Professional liability costs and managed care were both considered important contributing factors. Small proportions of specialists reported that they would definitely retire (7%) or relocate their practice out of state (4%) within the next 2 years; another third (32% and 29%, respectively) said they would likely do so. Forty-two percent of specialists have reduced or eliminated high-risk aspects of their practice, and 50% are likely to do so over the next 2 years. CONCLUSIONS: Our data suggest that claims of a "physician exodus" from Pennsylvania due to rising liability costs are overstated, but the malpractice situation is having demonstrable effects on the supply of specialist physicians in affected areas and their scope of practice, which likely impinges upon patients' access to care. PMID- 16244533 TI - Annals of surgery. PMID- 16244534 TI - A human factors analysis of technical and team skills among surgical trainees during procedural simulations in a simulated operating theatre. AB - BACKGROUND: High-risk organizations such as aviation rely on simulations for the training and assessment of technical and team performance. The aim of this study was to develop a simulated environment for surgical trainees using similar principles. METHODS: A total of 27 surgical trainees carried out a simulated procedure in a Simulated Operating Theatre with a standardized OR team. Observation of OR events was carried out by an unobtrusive data collection system: clinical data recorder. Assessment of performance consisted of blinded rating of technical skills, a checklist of technical events, an assessment of communication, and a global rating of team skills by a human factors expert and trained surgical research fellows. The participants underwent a debriefing session, and the face validity of the simulated environment was evaluated. RESULTS: While technical skills rating discriminated between surgeons according to experience (P = 0.002), there were no differences in terms of the checklist and team skills (P = 0.70). While all trainees were observed to gown/glove and handle sharps correctly, low scores were observed for some key features of communication with other team members. Low scores were obtained by the entire cohort for vigilance. Interobserver reliability was 0.90 and 0.89 for technical and team skills ratings. CONCLUSIONS: The simulated operating theatre could serve as an environment for the development of surgical competence among surgical trainees. Objective, structured, and multimodal assessment of performance during simulated procedures could serve as a basis for focused feedback during training of technical and team skills. PMID- 16244535 TI - Surgical simulation: a vital part of our future. PMID- 16244536 TI - Primary hyperparathyroidism, cognition, and health-related quality of life. AB - OBJECTIVE: To provide a rigorous and critical review of studies in which formal neuropsychological (NP) testing and measurement of health-related quality of life (HRQL) were conducted pre- and post-parathyroidectomy for primary hyperparathyroidism (PHPT). These data contribute to the discussion on the utility of surgical intervention for nonclassic PHPT. SUMMARY BACKGROUND DATA: PHPT is a complex endocrinopathy involving calcium metabolism and a potent hormone made by the parathyroid glands. Approximately 1.5% of Americans age 65 years and older, representing more than 3.9 million people, have PHPT, and the prevalence in postmenopausal women is estimated at 3.4%. Current National Institutes of Health guidelines for curative, surgical intervention of PHPT exclude 80% of patients with hyperparathyroid disease who have subjective neurobehavioral and physical symptoms that affect the quality of their lives. METHODS: An electronic search was conducted of prospective studies in which cognitive functioning was measured with formal NP tests and HRQL was measured with valid and reliable instruments before and following parathyroidectomy for PHPT. RESULTS: : In studies conducted pre- and post-parathyroidectomy for PHPT, 6 small studies of cognitive functioning report inconsistent findings; however, 7 well-designed studies of HRQL report improvement across multiple domains following surgery. CONCLUSIONS: Surgical treatment of PHPT is a viable option for patients with laboratory diagnosed, "nonclassic" PHPT. Formal NP testing and evaluation of HRQL are useful tools that may assist physicians in choosing whom to refer for parathyroidectomy. Further longitudinal study of NP functioning and HRQL in patients with laboratory diagnosed PHPT is warranted. PMID- 16244537 TI - Ultrasonically assisted retrohepatic dissection for a liver hanging maneuver. AB - OBJECTIVE: To establish a safer and technically easier retrohepatic dissection for the liver hanging maneuver with the assistance of intraoperative ultrasound (IOUS). SUMMARY BACKGROUND DATA: The liver hanging maneuver described by Belghiti et al is an innovative suspending technique of the liver and is useful in difficult major right hepatectomies or in donor operations for living donor liver transplantation. The most important complication of this procedure is injury to the short hepatic veins and subsequent massive bleeding with an incidence of 4% to 6%. METHODS: After the cranial dissection of the suprahepatic inferior vena cava (IVC) between the middle and left hepatic veins, a long light curved Kelly clamp is inserted from the caudal edge behind the caudate lobe and passed cranially along the anterior midline of the IVC. On the midway of the dissection, the proper hepatic vein draining the caudate lobe (PrCV) is visualized. A safe dissection path is confirmed by IOUS, identifying the position of the clamp tip, PrCV, and the caudal end of the cranial retrohepatic dissection. When IOUS shows that the clamp tip has reached the caudal end of the cranial dissection, the operator can feel the clamp tip with his/her finger and the retrohepatic dissection is completed. RESULTS: From September 2003 to July 2004, 50 donor operations were performed for adult living donor liver transplantation. Retrohepatic dissection was feasible in 40 cases (80%). Of these, a US-assisted retrohepatic dissection was performed in 34 donors. PrCVs were visualized by IOUS in 48 donors (96%). The location of these PrCVs varied significantly (60 degrees 175 degrees from the right edge of IVC), and there were no distinct landmarks for identifying the location of PrCVs and safe dissecting course (55 degrees -130 degrees ). IOUS found that the dissecting clamp was heading to the PrCV in 3 cases and the direction of dissection was shifted to avoid injury. No substantial bleeding or no other complication related to retrohepatic dissection was encountered in any of the cases. CONCLUSIONS: With the aid of IOUS, the whole course of the blind dissection between the anterior surface of the IVC and the liver could be clearly visualized. IOUS could also identify the PrCV, the most dangerous point in the retrohepatic dissection. PMID- 16244539 TI - Efficacy of sacral nerve stimulation for fecal incontinence: results of a multicenter double-blind crossover study. AB - BACKGROUND AND AIMS: This is the first double-blind multicenter study examining the effectiveness of sacral nerve stimulation in a significant number of fecally incontinent patients. METHODS: A total of 34 consecutive patients (31 women), median age 57 years (range, 33-73 years), underwent sacral nerve stimulation for fecal incontinence. After implantation, 27 of 34 patients were randomized in a double-blind crossover design to stimulation ON or OFF for 1-month periods. While still blinded, the patients chose the period of stimulation (ON or OFF) that they had preferred. The mode of stimulation corresponding to the selected period was continued for 3 months (final period). Outcome measures were frequency of fecal incontinence and urgency episodes, delay in postponing defecation, score severity, feeling of improvement, preference for ON or OFF, quality of life, and manometric measurements. RESULTS: In the crossover portion of the study, the self reported frequency of fecal incontinence episodes was significantly reduced during the ON versus the OFF period (P = 0.03), and this symptomatic improvement was consistent: 1) with the patients feeling of greater improvement during the ON versus OFF period (P = 0.02); 2) with the significant preference of patients (P = 0.02) for the ON versus OFF period. In the final period of the study, the frequency of fecal incontinence episodes decreased significantly (P = 0.005) in patients with the stimulator ON. The ability to postpone defecation (P = 0.01), the score for symptom severity (P = 0.0004), and the quality of life (P < 0.05) as well as anal sphincter function significantly improved. CONCLUSIONS: The significant improvement in FI during the ON versus OFF period indicated that the clinical benefit of sacral nerve stimulation was not due to placebo. PMID- 16244538 TI - Growth hormone, glutamine, and an optimal diet reduces parenteral nutrition in patients with short bowel syndrome: a prospective, randomized, placebo controlled, double-blind clinical trial. AB - OBJECTIVE: To determine if growth hormone (GH) and glutamine (Gln) might allow for a reduction in parenteral nutrition (PN) in individuals with short bowel syndrome. BACKGROUND DATA: Following massive intestinal resection, patients frequently sustain severe nutrient malabsorption and are dependent on PN for life. GH treatment with or without Gln might allow for a reduction in PN. METHODS: A prospective, double-blind, randomized, placebo-controlled clinical trial performed in 41 adults dependent on PN. Following screening, patients were admitted to an in-house facility for 6 weeks. After 2 weeks of stabilization and dietary optimization, patients were randomized to one of 3 treatment arms (1:2:2 ratio): oral Gln (30 g/day) + GH placebo (control group, n = 9), Gln placebo + GH (0.1 mg/kg per day, n = 16), or Gln + GH (n = 16). Standard criteria based on clinical and laboratory measurements were followed to determine PN volume and content. After 4 weeks of treatment, patients were discharged and monitored; GH and GH placebo were discontinued, but the diet with Gln or Gln placebo was continued for 3 months. RESULTS: Patients receiving GH + Gln placebo + diet showed greater reductions in PN volume (5.9 +/- 3.8 L/wk, mean +/- SD), PN calories (4338 +/- 1858 calories/wk), and PN infusions (3 +/- 2 infusions/wk) than corresponding reductions in the Gln + diet group (3.8 +/- 2.4 L/wk; 2633 +/- 1341 calories/wk; 2 +/- 1 infusions/wk, P < 0.05). Patients who received GH + Gln + diet showed the greatest reductions (7.7 +/- 3.2 L/wk; 5751 +/- 2082 calories/wk; 4 +/- 1 infusions/wk, P < 0.001 versus Gln + diet). At the 3-month follow-up, only patients who had received GH + Gln + diet maintained significant reductions in PN (P < 0.005) compared with the Gln + diet. CONCLUSIONS: Treatment with GH + diet or GH + Gln + diet initially permitted significantly more weaning from PN than Gln + diet. Only subjects receiving GH + Gln + diet maintained this effect for at least 3 months. PMID- 16244540 TI - Fibrin sealant versus mechanical stapling for mesh fixation during endoscopic extraperitoneal inguinal hernioplasty: a randomized prospective trial. AB - OBJECTIVE: To compare the clinical outcome of simultaneous bilateral endoscopic totally extraperitoneal inguinal hernioplasty (TEP) using fibrin sealant (FS) and mechanical stapling for prosthetic mesh fixation. SUMMARY BACKGROUND DATA: Similar efficacy of FS and mechanical stapling for mesh fixation has been demonstrated in a swine model, but no clinical trial has been conducted to compare the outcomes of TEP using these 2 fixation devices. FS adheres the prosthetic mesh without causing injury to the underlying structures. Whether the application of FS improves early postoperative outcomes, namely, reduction of postoperative pain and seroma formation, has not been examined. PATIENTS AND METHODS: Between July 2002 and February 2004, a total of 93 patients with 186 inguinal hernias who underwent bilateral TEP were randomized to have mesh fixation by either FS (n = 46) or mechanical stapling (n = 47). The primary endpoints were severity of pain, analgesic requirement, and incidence of seroma. Secondary endpoints were length of hospital stay, number of days required to resume normal outdoor activities and work, recurrence rate, and incidence of chronic pain. RESULTS: The 2 groups were comparable in age, sex, and types of hernia. TEP were successfully performed in all patients. The FS group consumed significantly less analgesics compared with that of the staple group (P = 0.034). There was no significant difference in the postoperative pain score at rest and on coughing from the day of operation to postoperative day 6 between the groups. The incidence of seroma was significantly higher in the FS group (17.4%) than the staple group (5.3%) (P = 0.009). Length of hospital stay and time taken to resume normal activities and work were comparable between the 2 groups. With a median follow-up of 1.2 years, no recurrent hernia has been detected in either group, but the incidence of chronic pain in the staple group (20.0%) was higher than that of the FS group (13.2%) (P = 0.418). CONCLUSIONS: This randomized prospective clinical trial demonstrated a significant reduction of analgesic consumption by using FS for mesh fixation during bilateral TEP, but it was associated with an increased incidence of postoperative seroma. PMID- 16244541 TI - Modulation of E-cadherin by hepatocyte growth factor induces aggressiveness of gastric carcinoma. AB - OBJECTIVE: Hepatocyte growth factor (HGF) is well known as a scatter factor because it can disperse cells. E-cadherin is a protein that plays a main role in the establishment of cell-cell adhesion. This study focused on the role of HGF on the expression and distribution of E-cadherin. Furthermore, we found induction of aggressiveness of gastric carcinoma by modulation of E-cadherin by HGF. MATERIALS AND METHODS: Tumor tissues from 50 patients with gastric carcinoma were evaluated for the expression of HGF, its receptor c-Met, and E-cadherin. Western blot analysis and invasion assay were performed to confirm the role of HGF on the modulation of E-cadherin using human gastric cancer cell lines. RESULTS: Seventy eight percent of the gastric carcinoma tissues showed overexpression of c-Met. E cadherin expression was found in 86%, which could be further classified as membranous type (52%) or nonmembranous type (48%). The levels of HGF in tumor tissues increased significantly according to the tumor progression. The levels of HGF in tumors with nonmembranous type E-cadherin expression were significantly higher than those in tumors with membranous expression. A striking morphologic change from epithelial shape to fibroblastic shape was observed in SNU-16 cells after 3 days' exposure to HGF, accompanied by down-regulation of functional E cadherin in the membrane. Treatment of the cells with HGF induced significant invasion into the matrigel. CONCLUSION: We can conclude that HGF can modulate the expression of E-cadherin in gastric carcinoma, which was accompanied by more aggressive phenotype. PMID- 16244542 TI - Histomorphologic tumor regression and lymph node metastases determine prognosis following neoadjuvant radiochemotherapy for esophageal cancer: implications for response classification. AB - OBJECTIVE: We sought to quantitatively and objectively evaluate histomorphologic tumor regression and establish a relevant prognostic regression classification system for esophageal cancer patients receiving neoadjuvant radiochemotherapy. PATIENTS AND METHODS: Eighty-five consecutive patients with localized esophageal cancers (cT2-4, Nx, M0) received standardized neoadjuvant radiochemotherapy (cisplatin, 5-fluorouracil, 36 Gy). Seventy-four (87%) patients were resected by transthoracic en bloc esophagectomy and 2-field lymphadenectomy. The entire tumor beds of the resected specimens were evaluated histomorphologically, and regression was categorized into grades I to IV based on the percentage of vital residual tumor cells (VRTCs). A major response was achieved when specimens contained either less than 10% VRTCs (grade III) or a pathologic complete remission (grade IV). RESULTS: Complete resections (R0) were performed in 66 of 74 (89%) patients with 3-year survival rates of 54% +/- 7.05% for R0-resected cases and 0% for patients with incomplete resections or tumor progression during neoadjuvant therapy (P < 0.01). Minor histopathologic response was present in 44 (59.5%) and major histopathologic response in 30 (40.5%) tumors. Significantly different 3-year survival rates (38.8% +/- 8.1% for minor versus 70.7 +/- 10.1% for major response) were observed. Univariate survival analysis identified histomorphologic tumor regression (P < 0.004) and lymph node category (P < 0.01) as significant prognostic factors. Pathologic T category (P < 0.08), histologic type (P = 0.15), or grading (P = 0.33) had no significant impact on survival. Cox regression analysis identified dichotomized regression grades (minor and major histomorphologic regression, P < 0.028) and lymph node status (ypN0 and ypN1, P < 0.036) as significant independent prognostic parameters. A 2-parameter regression classification system that includes histomorphologic regression (major versus minor) and nodal status (ypN0 versus ypN1) was established (P < 0.001). CONCLUSIONS: Histomorphologic tumor regression and lymph node status (ypN) were significant prognostic parameters for patients with complete resections (R0) following neoadjuvant radiochemotherapy for esophageal cancer. A regression classification based on 2 parameters could lead to improved objective evaluation of the effectiveness of treatment protocols, accuracy of staging and restaging modalities, and molecular response prediction. PMID- 16244543 TI - Crohn's disease patients carrying Nod2/CARD15 gene variants have an increased and early need for first surgery due to stricturing disease and higher rate of surgical recurrence. AB - OBJECTIVE: To study the predictive value of Nod2/CARD15 gene variants along with disease phenotypic characteristics for requirement of initial surgery and for surgical recurrence in Crohn's disease (CD). SUMMARY BACKGROUND DATA: Nod2/CARD15 gene variants play an important role in the susceptibility to CD. Studies of genotype-phenotype relationship suggest that these variants are associated with development of intestinal strictures. Preliminary reports analyzing the association between these variants and need for surgery have produced inconsistent results. METHODS: A total of 170 CD patients were included prospectively in the study and followed up regularly for a mean of 7.4 +/- 6.1 years. Clinical characteristics of CD, time and indication for surgery, and recurrence were registered. Nod2/CARD15 gene variants were determined by DNA sequencing analysis. RESULTS: Surgery for stricturing disease was significantly more frequent in patients with Nod2/CARD15 variants in the univariate analysis (odds ratio [OR], 3.63; 95% confidence interval [CI], 1.42-9.27), and it was required at an earlier time (P = 0.004). Only Nod2/CARD15 variants (OR, 3.58; 95% CI, 1.21-10.5) and stricturing phenotype at diagnosis of CD (OR, 9.34; 95% CI, 2.56-33.3) were independent predictive factors of initial surgery for stricturing lesions in the multivariate analysis. Among 70 patients that required surgery, postoperative recurrence was also more frequent in patients with Nod2/CARD15 variants in the univariate and multivariate analysis (OR, 3.29; 95% CI, 1.13 9.56), and reoperation was needed at an earlier time (P = 0.03). CONCLUSION: Nod2/CARD15 variants are associated with early initial surgery due to stenosis and with surgical recurrence in Crohn's disease. Patients with these variants could benefit from preventive and/or early therapeutic strategies. PMID- 16244544 TI - The accuracy of ultrasound, stereotactic, and clinical core biopsies in the diagnosis of breast cancer, with an analysis of false-negative cases. AB - OBJECTIVE: Preoperative core biopsy in breast cancer is becoming the standard of care. The aim of this study was to analyze the various methods of core biopsy with respect to diagnostic accuracy and to examine the management and outcome of those patients with false-negative biopsies. METHODS: All patients undergoing core biopsy for breast abnormalities over a 5-year period (1999-2003) were reviewed. The accuracy rates for each method of core biopsy, the histologic agreement between the core pathology and subsequent excision pathology, and the length of follow-up for cases of benign disease were studied. Patients whose biopsies were benign but who were subsequently diagnosed with cancer underwent detailed review. RESULTS: There were 2427 core biopsies performed over the 5-year period, resulting in a final diagnosis of cancer in 1384 patients, benign disease in 954 patients, and atypical disease in 89 patients. Biopsy type consisted of 1279 ultrasound-guided cores, 739 clinically guided cores, and 409 stereotactic guided cores. The overall false-negative rate was 6.1%, with specific rates for ultrasound-, clinical-, and stereotactic-guided cores of 1.7%, 13%, and 8.9%, respectively. False-negative biopsies occurred in 85 patients, and in 8 of these patients the diagnosis was delayed by greater than 2 months. In all other false negative cases, "triple assessment" review allowed prompt recognition of discordant biopsy results and further evaluation. CONCLUSION: Ultrasound guidance should be used to perform core biopsies in evaluating all breast abnormalities visible on ultrasound. Adherence to principles of triple assessment following biopsy allows for early recognition of the majority of false-negative cases. PMID- 16244545 TI - Follicular thyroid carcinoma: the role of histology and staging systems in predicting survival. AB - OBJECTIVE: To evaluate the risk factors including tumor histomorphology for survival specific to follicular thyroid carcinoma (FTC) and to apply commonly employed staging systems in predicting survival for patients with FTC. SUMMARY BACKGROUND DATA: FTC is usually analyzed collectively with papillary thyroid carcinoma (PTC) in risk group analysis. Risk factors and risk group analysis are important in the management of patients with FTC, although current published therapeutic guidelines call for total thyroidectomy followed by radioactive iodine (I) ablation for all FTC patients. METHODS: Over a 40-year period, 156 patients surgically treated for FTC with an average follow-up of 14.4 years were retrospectively studied after histologic reclassification according to the type and degree of invasiveness of the tumor. Potential risk factors for survival were calculated using multivariate analysis, and the prognostic accuracy of AMES risk group stratification, UICC/AJCC pTNM staging, Degroot classification, and MACIS scoring schemes in predicting survival was compared. RESULTS: Seventeen (11%) patients had distant metastases at presentation, and bilateral thyroid resection was performed for 131 (84%) patients. Seventeen (11%) patients died of recurrent or metastatic disease. The overall and cancer-specific survival (CSS) rates at 10 years were 79% and 88%, respectively. None of the patients with minimally invasive (n = 49) or angioinvasive (n = 23) carcinomas died compared with 17 of 84 patients with widely invasive carcinomas (P = 0.0007). Using the Cox proportional hazards model, old age, the presence of distant metastases, and incomplete tumor excision were independent prognostic factors for survival. For patients who underwent curative treatment, old age and widely invasive carcinoma were risk factors for poor survival. All staging systems studied accurately predicted CSS, and the pTNM UICC/AJCC staging system yielded the best prognostic information. CONCLUSIONS: Commonly adopted staging systems can be applied specifically to patients with FTC. The distinction of FTC in minimally invasive and widely invasive carcinoma based on the extent of invasiveness rather than vascular invasion is important in identifying low-risk FTC patients for a more conservative management. PMID- 16244546 TI - Simultaneous pancreas-kidney transplantation from donation after cardiac death: successful long-term outcomes. AB - OBJECTIVE: The outcomes of simultaneous pancreas-kidney (SPK) transplantation with donor organs procured from donation after cardiac death (DCD) are compared with transplants performed with donor organs recovered from donation after brain death (DBD). SUMMARY BACKGROUND DATA: Concerns exist regarding the utilization of pancreata obtained from DCD donors. While it is known that DCD kidneys will have a higher rate of DGF, long-term functional graft survival data for DCD pancreata have not been reported. METHODS: A retrospective review of all DCD SPK transplants performed at a single center was undertaken. RESULTS: Patient, pancreas, and kidney survival at 5 years were similar between DCD and DBD organs. Pancreas function and outcomes were indistinguishable between the 2 modes of procurement. As expected, the DCD kidneys had an elevated rate of DGF, which had no significant long-term clinical impact. CONCLUSION: SPK transplantation using selected DCD donors is a safe and viable method to expand the organ pool for transplantation. PMID- 16244547 TI - Donation after cardiac death: the University of Wisconsin experience with liver transplantation. AB - OBJECTIVE: To determine whether the outcomes of liver transplantation (LTx) from donation after cardiac death (DCD) donors are equivalent to those from donation after brain death (DBD) donors. SUMMARY BACKGROUND DATA: Because of the significant donor organ shortage, more transplant centers are using livers recovered from DCD donors. However, long-term, single-center outcomes of liver transplantation from DCD donors are limited. METHODS: From January 1, 1993, to July 31, 2002, 553 liver transplants were performed from DBD donors and 36 were performed from DCD donors. Differences in event rates between the groups were compared with Kaplan-Meier estimates and the log-rank test. Differences in proportion and differences of means between the groups were compared with Fisher exact test and the Wilcoxon rank sum test, respectively. RESULTS: Mean warm ischemic time at recovery in the DCD group was 17.8 +/- 10.6 minutes. The overall rate of biliary strictures was greater in the DCD group at 1 year (33% versus 10%) and 3 years (37% versus 12%; P = 0.0001). The incidence of hepatic artery thrombosis, portal vein stenosis/thrombosis, ischemic-type biliary stricture (ITBS), and primary nonfunction were similar between groups. However, the incidence of both hepatic artery stenosis (16.6% versus 5.4%; P = 0.001) and hepatic abscess and biloma formation (16.7% versus 8.3%; P = 0.04) were greater in the DCD group. Trends toward worse patient and graft survival and increased incidence of ITBS were seen in DCD donors greater than 40 years compared with DCD donors less than 40 years. Overall patient survival at 1 year (DCD, 80%; versus DBD, 91%) and 3 years (DCD, 68%; versus DBD, 84%) was significantly less in the DCD group (P = 0.002). Similarly, graft survival at 1 year (DCD, 67%; versus DBD, 86%) and 3 years (DCD, 56%; versus DBD, 80%) were significantly less in the DCD group (P = 0.0001). CONCLUSIONS: Despite similar rates of primary nonfunction, LTx after controlled DCD resulted in worse patient and graft survival compared with LTx after DBD and increased incidence of biliary complications and hepatic artery stenosis. However, overall results of LTx after controlled DCD are encouraging; and with careful donor and recipient selection, LTx after DCD may successfully increase the donor liver pool. PMID- 16244548 TI - Single-center experience with liver transplantation from controlled non heartbeating donors: a viable source of grafts. AB - BACKGROUND: To increase the number of livers available for transplantation a non heartbeating donor (NHBD) liver transplant program was started after obtaining hospital ethical committee approval. METHODS: Controlled donors with a warm ischemia of <30 minutes were considered. A 5-minute stand-off period was observed from asystole to skin incision. A super-rapid technique was used for the retrieval. Methods used to assess the suitability for transplantation included liver function tests, morphologic and histologic assessment, and hepatocyte viability testing. RESULTS: Sixty livers were retrieved from NHBDs. Of these, 33 were judged suitable for transplantation. Of these one was exported and transplanted, and one could not be matched to a suitable recipient. A further 27 were not used because of liver appearance in 21, prolonged hypoxia and hypotension in 4, poor perfusion in 1, and donor malignancy in 1. Mean donor age was 39.4 years (range, 0.75-67 years). Causes of death were head trauma in 10 donors, intracranial bleed in 24, and anoxic/ischemic brain injury in 26. Mean warm ischemia time was 14.7 minutes (range, 7-40 minutes). Thirty-two patients were transplanted (one split liver), and the mean age of the recipients was 38.4 years (range, 0.7-72 years). All grafts had good early function except one right lobe split. There were 4 deaths resulting from ischemic brain injury, chronic rejection, biliary sepsis, and multiorgan failure following retransplantation for primary nonfunction. Overall patient and graft survival is 87% and 84%, respectively, at a median follow-up of 15 months. CONCLUSIONS: Early results suggest that controlled NHBDs are a significant new source of grafts, but careful donor selection and short cold ischemia are mandatory. PMID- 16244551 TI - Problem of portal venous thrombosis after splenectomy. PMID- 16244549 TI - French women from multiplex abdominal aortic aneurysm families should be screened. AB - BACKGROUND: Multiplex abdominal aortic aneurysm families (MAAAFs) (> or =1 subject plus the proband) represent 1% to 34% of abdominal aortic aneurysm (AAA), but the percentage in France is unknown. METHOD: The MAAAF rate was retrospectively defined by analysis of 3 groups: 72 of 104 consecutive individuals undergoing AAA surgery during 1994, 24 of 53 women and 35 of 76 men with giant (> or =9 cm) AAA operated on during 1986 to 1994. MAAAF characteristics were determined based on 10 families issued from these 3 groups and 34 others identified nationwide. Data were obtained from a standardized questionnaire for probands and relatives, detailed pedigrees of each family, and computed tomography (CT) scans without contrast medium of the aorta and lower limb arteries for first-degree relatives > or =40-year-of age. RESULTS: The MAAAF rate was 4.2% for the consecutive-surgery patients (proband M/F ratio, 17:1; mean age at surgery, 68.5 +/- 8.5 years). CT detected no additional AAA among them (screened individuals M/F ratio, 0.63; mean age, 54.0 +/- 11.2 years). MAAAF rates were 8.3% and 14.3% for the women's and giant-AAA groups with CT screening, respectively. Characteristics were investigated in 104 affected subjects from 44 MAAAFs: female relatives were more often affected than probands (P < 0.025). Compared with men, affected female relatives were significantly older at diagnosis and surgery (P < 0.05 and P < 0.02, respectively), as were affected women (P < 0.02 and P < 0.01, respectively). CT scan screening identified significantly more AAA and abdominal aortic dilatations among the 44 MAAAFs than the consecutive-surgery group (5 and 4, respectively; P < 0.001). CONCLUSION: Although the MAAAF rate seems low in France, women from MAAAF were affected more often and later, suggesting that they should be screened. PMID- 16244552 TI - Laparoscopic left colectomy and obese patients. PMID- 16244557 TI - [Obsessive compulsive disorders associated with Gilles de la Tourette syndrome]. PMID- 16244553 TI - Are there curative options to peritoneal carcinomatosis? PMID- 16244556 TI - New technique for liver resection using heat coagulative necrosis. PMID- 16244558 TI - [Obituary for Professor Jean-Marie Warter]. PMID- 16244559 TI - [Spatio-temporal dynamics of neuronal networks in partial epilepsy]. AB - INTRODUCTION: The anatomo-functional organization of partial drug-resistant epilepsies is the subject of much current research aiming at better understanding these pathologies and improving their treatment. The work carried out by our team on the study of intracerebral recording falls within this category of research. The objectives are to identify the neural networks involved in the generation of paroxysmal activity and to understand their spatio-temporal dynamics, in order to be able in the long term to propose targeted therapeutic approaches likely to "control" these networks. STATE OF ART: The traditional concept of epileptic "focus" must nowadays be replaced by a more complex model taking into account potential interactions within the neural networks involved in the seizure. Indeed, during partial seizures, involved cerebral structures are the site of characteristic oscillations which may be synchronized or on the contrary transiently desynchronized. These epileptic rhythms may disturb the physiological rhythms underlying normal cognitive processes; these cognitive processes may thus be impaired in partial epilepsy, even those remote from the site of origin of the discharge. In this article we describe a model of organization of human partial seizures, through characterization of the relationships ("synchrony") between intracerebral signals recorded in the involved structures. We propose that seizures are generated in an initial network of highly epileptogenic brain structures (epileptogenic zone network, EZN) whose activity is synchronized; this activity is then transiently desynchronized with the appearance of fast oscillations. During a second ictal phase, other cortical and subcortical structures are the seat of slower rhythmic modifications that are synchronized (propagation network, PN). The emergence of a particular clinical semiology in the course of the seizure depends on these phenomena which can in certain cases "mimic" a normal cerebral process or on the contrary provoke a major rupture in normal cerebral functioning. CONCLUSIONS: These studies contribute to improvement in our knowledge of the neural networks involved in partial epilepsies. In the future, this type of research may contribute to the development of specific treatments that target certain pathophysiological mechanisms involved in seizure generation. PMID- 16244560 TI - [The use of intravenous immunoglobulins in neurology]. AB - INTRODUCTION: In the past decade, intravenous immunoglobulins (IVIG) have been widely used and their administration has grown throughout the world. The current indications of IVIG in neurological diseases are discussed on the basis of the passed and current trials. Unlike other immuomodulatory agents, IVIG are well tolerated and have very few side effects and a good viral safety. STATE OF ART: There is clinical evidence, based on controlled trials, for the effectiveness of IVIG in Guillain-Barre syndrome, chronic inflammatory demyelinating polyneuropathy and multifocal neuropathy with conduction blocks. In myasthenia gravis, the IVIG are effective especially in myasthenic crisis, but their synergistic effect with other treatments, the steroid sparing effect, and their long-term effect are unknown. These issues need to be addressed in further controlled clinical trials. In dermatoploymyositis, IVIG are reserved for steroid resistant patients. There is actually no support or no significant clinical benefit for the routine use of IVIG in other neurological diseases. PERSPECTIVES: Further controlled trials are warranted to assess the quality of life, the dose finding effect and their long-term efficacy in order to improve clinical practices. CONCLUSION: Routine use of IVIG should be reserved for diseases in which positive controlled trials are available. For the remaining dysimmune diseases, IVIG should be assess in comparison with the other available therapies, taking into consideration the age of the patients, the safety of the IVIG and, in our country, the economic aspect. PMID- 16244561 TI - [Gilles de la Tourette syndrome: a self-administered assessment questionnaire]. AB - INTRODUCTION: An association of patients with Gilles de la Tourette syndrome enabled us to gather a large body of information regarding the disease manifestations, and patient-perceived consequences. METHOD: 350 questionnaires were sent to patients belonging to the AFSGT (French Association of Patients Suffering from Gilles de la Tourette Syndrome). 187 responses were received (53 percent). The patients were divided into four groups: those with motor tics, vocal tics, complex tics and complex tics with coprolalia. This last group corresponds to the DSM IV definition of "Tourette Disorder". The questions were grouped in five sections: simple manifestations, complex manifestations, family study, treatment and psycho-affective perception (social and in the context of schooling). RESULTS: The study of the simple manifestations of the disorder revealed the homogeneity of the four groups with an age of onset at on average 7 years and a male-to-female ratio of 3.5. The first signs of the disorder are motor tics of the face and neck, and the disorder shows a variable and fluctuating course characterized by periods of decreased or absent symptoms. Familial cases (58 percent) are found in all four groups. The complex signs included in part of behaviors corresponding to the definition of tics: sudden, brusque, repetitive, varied, escape despite efforts to repress them and reappearance more intensely after a period of conscious control. The complex signs also consisted of accompanying factors such as agitation, need to organize, classify or count. Treatments have been of limited success and a significant number of patients have abandoned treatment entirely. Our study demonstrates that this condition seriously affects the daily life of patients, including family and social relations, schooling and occupational life. No patients suffering from transient tics responded to our survey, but such tics were reported in family members. CONCLUSION: Overall, the condition is considered to be single family of disorders, despite the broad phenotypic spectrum, from transitory cases by children to very severe forms. Escape despite efforts to repress tics and the rebound after control tics is characteristic of the Georges Gilles de la Tourette syndrome. PMID- 16244562 TI - [False recognition of faces associated with fronto-temporal dementia with prosopagnosia]. AB - INTRODUCTION: The association of prosopagnosia and false recognition of faces is unusual and contributes to our understanding of the generation of facial familiarity. METHOD: A 67-year-old man with a left prefrontal traumatic lesion, developed a temporal variety of fronto-temporal dementia (semantic dementia) with amyotrophic lateral sclerosis. Cerebral imagery demonstrated a bilateral, temporal anterior atrophy predominating in the right hemisphere. The main cognitive signs consisted in severe difficulties to recognize faces of familiar people (prosopagnosia), associated with systematic false recognition of unfamiliar people. RESULT: Neuropsychological testing indicated that the prosopagnosia probably resulted from the association of an associative/mnemonic mechanism (inability to activate the Face Recognition Units (FRU) from the visual input) and a semantic mechanism (degradation of semantic/biographical information or deconnexion between FRU and this information). At the early stage of the disease, the patient could activate residual semantic information about individuals from their names, but after a 4-year course, he failed to do so. This worsening could be attributed to the extension of the degenerative lesions to the left temporal lobe. Familiar and unfamiliar faces triggered a marked feeling of knowing. False recognition concerned all the unfamiliar faces, and the patient claimed spontaneously that they corresponded to actors, but he could not provide any additional information about their specific identities. The coexistence of prosopagnosia and false recognition suggests the existence of different interconnected systems processing face recognition, one intended to identification of individuals, and the other producing the sense of familiarity. Dysfunctions at different stages of one or the other of these two processes could result in distortions in the feeling of knowing. CONCLUSION: From this case and others reported in literature, we propose to complete the classical model of face processing by adding a pathway linked to limbic system and frontal structures. This later pathway could normally emit signals for familiarity, essentially autonomic, in response to the familiar faces. These signals, primitively unconscious, secondly reach consciousness and are then integrated by a central supervisor system which evaluates and verifies identity-specific biographical information in order to make a decision about the sense of familiarity. PMID- 16244563 TI - [A family with exercise-induced paroxysmal dystonia and childhood absence epilepsy]. AB - INTRODUCTION: The boundary between epilepsy and paroxysmal dyskinesia appears to be less easy to delineate than previously believed. Reports of families showing both phenomena suggest a shared pathophysiology. PATIENTS AND METHOD: A new family with autosomal dominant exercise-induced paroxysmal dystonia is reported. RESULTS: Two family members also had childhood absence epilepsy, and one of them suffered from acute transient hemiplegia at age 10. CONCLUSION: The association of epilepsy and paroxysmal dyskinesia has been rarely reported in the literature, and several loci have been identified. Absence epilepsy and exercise-induced paroxysmal dystonia appear to be very uncommon, although some reports mentioned the association in sporadic and familial cases. The involvement of ion channel genes in several transient neurological disorders supports the hypothesis of a common pathophysiological process underlying both the childhood absence seizure and the later paroxysmal dystonia. PMID- 16244564 TI - [Non-Hodgkin's malignant lymphoma of the peripheral nervous system: clinicopathological correlations in ten patients]. AB - INTRODUCTION: Identifying tumor infiltration or compression in patients with non Hodgkin's malignant lymphoma presenting peripheral neuropathy can be a difficult task. METHODS: We collected a series of patients with peripheral neuropathy with demonstrated lymphomatous infiltration or compression managed between October 1977 and October 2001 to search for clinico-pathological correlations. RESULTS: Ten cases were reviewed. Neurological manifestations were the inaugural symptom of the disease in 7 patients. Clinical presentations included 5 focal (3 cranial nerve palsies, 2 brachial radiculopathies) and 5 diffuse neuropathies (3 polyradiculoneuropathies, 1 polyneuropathy and 1 mononeuritis multiplex). The mechanisms of peripheral nerve involvement were classified into lymphomatous meningoradiculitis (5 cases), involvement of cranial nerves or spinal roots in their extraneuraxial course (3 cases) and infiltration of distal peripheral nerves (2 cases). Four long lasting survivals after treatment were observed. CONCLUSIONS: Prognosis depends much more on the haematological disease than on the neurological symptoms or tumor location. PMID- 16244565 TI - [Crossed anarthria and a dissociated lateralisation of language]. AB - INTRODUCTION: Crossed anarthria cases are uncommon and rather old. OBSERVATION: We report the case of a right-handed 55-year-old man who presented crossed pure anarthria due to a hemorrhage in the premotor cortex (feet of F1 and F2) and in the high part of Pierre-Marie's quadrangle. CONCLUSION: The study of different tasks (articulation, verbal fluency, direct object word-generation from a verb) showed a dissociated lateralisation of his language. Lexico-semantic and grammatical tasks are processed in the left hemisphere. Articulation programming occurs in the right hemisphere. PMID- 16244566 TI - [Systemic Candida albicans infection with neuromeningeal complication in a premature infant]. AB - INTRODUCTION: Systemic maternal-fetal Candida albicans infections are uncommon diseases with a poor outcome. An associated cerebromeningeal infection increases morbidity. We present a case of neuromeningeal candidiasis following systemic neonatal infection in a premature infant. Management and therapeutic difficulties are outlined. OBSERVATION: The patient was a male infant born preterm at 30 weeks gestation. During his first week of life, he developed a systemic infection with an associated symptomatic hydrocephalus. Systemic candidaisis with neuromeningeal complication was diagnosed five weeks later. Despite treatment including cerebrospinal fluid (CSF) shunting and antimycotic medications (flucytosin and amphotericin B), the candidal infection did not resolve. Infectious and mechanical complications of the CSF drainage were treated by several surgical interventions during the following months. At 10 months of life, there was clinical and laboratory evidence of active persistent neuromeningeal candidaisis. Finally, candidal infection was eradicated with intravenous administration of fluconazole. After five year follow-up, the intellectual and psychological status of the patient was quite satisfactory, and no neurological deficits were found on clinical examination. DISCUSSION: Management of neuromeningeal candidaisis in premature infants is a challenging task particularly because of delayed diagnosis. Candida infection should routinely be suspected in cases of systemic infection with neurological impairment in premature infants. Fluconazole may constitute an efficient therapeutic option. PMID- 16244567 TI - [Cerebral vasculitis secondary to Varicella-Zoster virus infection]. AB - INTRODUCTION: Central nervous system infection by the varicella-zoster virus (VZV) can be responsible for myelitis, meningitis, ventriculitis and large and small-vessels encephalitis. CASE REPORT: We report the case of a 57-year-old-man hospitalized for deteriorating general health. Physical examination revealed likely encephalitis associated with headache without meningeal syndrome. Successive cerebral MRIs showed bilateral necrosis of the amygdaloid bodies and multiple deep and sub-cortical infarcts suggestive of vasculitis. Cerebral arteriography was normal. Three cerebral fluid examinations disclosed mononuclear pleiocytosis with few red blood cells. PCR analysis for VZV was only positive at the third time. DISCUSSION: The diagnosis of VZV encephalitis is difficult without the rash typical of zoster and because of the low sensitivity of PCR VZV in comparison with PCR HSV. CONCLUSION: In active viral disease, where the prognosis depends on early treatment, we highlight the usefulness of repeated PCR analysis and the search for antibodies in blood and cerebrospinal fluid. PMID- 16244568 TI - [Multiple strokes after chickenpox primo-infection in an adult]. AB - INTRODUCTION: Chickenpox is considered as a high risk factor for developing stroke in childhood, but descriptions in adult are exceptional (only three cases reported, to our knowledge). CASE REPORT: A 37-year-old man presented with a chickenpox eruption, followed by a right parietal and a left occipital infarcts, associated with multiple lacunae. There was no coagulation disorder, no hypertension or cardiovascular disorder. Cerebral angiography showed an irregular narrowing of the right internal parietal artery and vascular defects in right parietal and left occipital areas. The diagnosis of VZV-related vasculitis was evoked. White cell count, serology and VZV PCR were negative in the cerebrospinal fluid. Clinical improvement was observed after treatment by corticosteroids and aciclovir. CONCLUSION: Chickenpox is a rare cause of cerebral vasculitis. Involvement of both medium and small vessels was present here, contrary to other adult case reports in the literature. Hematogenous dissemination of the virus responsible for cerebral vasculitis seems to be the most probable pathophysiological mechanism. PMID- 16244569 TI - [Recurrent Miller-Fisher syndrome]. AB - INTRODUCTION: Miller-Fisher syndrome (MFS) is a rare auto-immune post-infectious syndrome, characterized by an ataxia, an ophthalmoplegia and a generalized areflexia. It is considered as a clinical variant of Guillain-Barre syndrome (GBS). MFS is correlated with the presence of anti-GQ1b antibodies, elevated cerebrospinal fluid (CSF) protein levels, presence of mostly sensitive electrophysiological abnormalities and for some authors central involvement with increased signal intensity of brainstem and cerebellum on MRI. Recurrent MFS is extremely rare with only 21 cases since the first description in 1970. CASE REPORT: A 54-year-old women presented MFS with two episodes in 19 years. Clinically, the first episode was a "classical" MFS, and the second an extensive MFS with tetraparesis and respiratory failure. CSF protein levels and cerebral MRI were normal. Anti-GQ1b antibodies were strongly positive and anti-GM1, anti GM2 antibodies were slightly positive, campylobacter jejuni serology was negative. Electromyography showed isolated sensory abnormalities in median nerves territory. CONCLUSION: We report a new case of recurrent MFS with unusual clinical, biological and electrophysiological features. PMID- 16244570 TI - [Hereditary neuropathy with liability to pressure palsy presenting with an acute inflammatory demyelinating polyneuropathy]. PMID- 16244571 TI - [Acute tetraparesis of cerebral origin]. AB - INTRODUCTION: Thrombolytic treatment in the early stage of ischemic cerebral attacks requires rapid confirmation of the diagnosis and topographic localization. Unusual clinical features can lead to misdiagnosis with the risk of delaying optimal therapeutic management. OBSERVATION: We report the cases of two patients who experienced acute tetraparesis without any associated encephalic sign, consistent with the diagnosis of spinal cord injury. Cervical magnetic resonance imaging (MRI) was normal. Conversely, cerebral MRI displayed in both cases bilateral hemispheric infarction. Two ischemic lesions were revealed in the territory of both anterior cerebral arteries in the first patient, while the second patient had a bilateral infarction in the posterior arms of both internal capsules. CONCLUSION: In case of tetraparesis, emergency spinal cord MRI should be performed to rule out neurosurgical etiologies and ischemia. If negative, cerebral MRI should be performed at the same time to look for early cerebral infarction in both hemispheres and determine the indication for thrombolysis. PMID- 16244572 TI - [Speech therapy for the dysarthria of Parkinson's disease]. AB - INTRODUCTION: Dysarthria is frequent in Parkinson's disease and impairs communication. STATE OF THE ART: Use of levodopa or surgical procedures does not significantly improve speech. Traditional speech therapy can lead to improvement of dysarthria, and intensive programs have recently showed substantial beneficial effects on parkinsonian dysarthria. PERSPECTIVES: More work is needed to evaluate speech treatment efficacy. CONCLUSION: Earlier and more frequent speech therapy in parkinsonian dysarthria patients may contribute to better quality of life. PMID- 16244573 TI - [The finding of incidental intracranial lesions with magnetic resonance imaging]. AB - INTRODUCTION: Incidental findings on brain MRI are not rare since the development of MRI as a diagnostic and clinical research tool. STATE OF THE ART: Prevalence of cerebral incidental finding, based on research volunteers participating in studies, is estimated from 1.7 to 4 percent. The most frequently detected lesions are intracranial tumors (meningioma, arachnoid cysts, neuro-epithelial tumor) and vascular malformations. PERSPECTIVES: A better knowledge of the spontaneous outcome of incidental findings would allow adequate management of these patients. CONCLUSION: Clinicians should be aware of the outcome of incidental findings, to reassure patients or trigger a work-up if necessary. PMID- 16244574 TI - [Consensus statement on severe dementia]. AB - Under the auspices of the French Society of Gerontology and Geriatrics, a multidisciplinary team including geriatritians, neurologists, epidemiologists, psychiatrists, pharmacologists and public health specialists developed a consensus on care for patients with severe dementia. They defined 21 recommendations for general practitioners, long-term care physicians and specialists based on knowledge available in 2005. At all stages of the disease, the objective of care is to improve as much as possible quality-of-life for the patient and his/her family, including a life project until the end of life. It is always possible to do something for these patients and their family: nutritional status, behavior disorders, and incapacities to deal with basic activities of daily life have to be taken in consideration. Resource allocation and proximity care have to be targeted. Research areas necessary to improve the care of patients with severe dementia has been selected. PMID- 16244576 TI - Colorectal cancer and hypercholesterolemia: review of current research. AB - AIM: In spite of ample research about a high level of cholesterol in the blood of patients with colorectal cancer (CRC), the relationship between factors causing hypercholesterolemia and factors leading to CRC development is not fully investigated. The purpose of this article is to provide a review of the current research about the risk factors leading to the development of hypercholesterolemia and CRC, and to show the relationship between these factors, hypercholesterolemia and CRC with the implication for CRC preventive and treatment practices. METHODS: A systematic search of MEDLINE and PUBMED databases between 1990 and 2005 was conducted to locate the studies that investigated the risk factors causing CRC and hypercholesterolemia. From among 255 studies found, 66 were selected that matched the following criteria for selection: (1) reported original research; (2) discussed at least one of the listed eight factors; (3) discussed hypercholesterolemia; and/or (4) discussed colon or rectum cancer. RESULTS: The studies were grouped according to four areas of research: (1) studies that explored the relationship between different factors and CRC incidences; (2) studies that investigated the relationship between different factors and CRC incidences and the role of mutations in causing CRC; (3) studies that looked at the factors causing hypercholesterolemia; and (4) studies that explored the relationship among the factors, hypercholesterolemia, and CRC development. A discussion of the studies is presented and the details related to the studies major aspects are summarized in 4 tables. CONCLUSION: The review has revealed a relationship between factors that can lead to the development of CRC and those that lead to hypercholesterolemia. Although the role of many individual risk factors is still controversial the analysis of their significance in combination might be important for diagnostic and development of the models for prediction of cancer occurrence. PMID- 16244577 TI - Hypercoagulant states in malignant lymphoma. AB - The incidence of severe complications, such as disseminated intravascular coagulation (DIC) in malignant lymphoma, differs between clinical stages and histological types of the disease, but they occur frequently in stage IV or natural killer (NK) cell lymphoma. Patients with stage IV or NK cell lymphoma exhibit abnormal thrombotic and hemostatic states. One of the mechanisms in DIC might involve elevated cytokine expression by lymphoma cells stimulating the expression of tissue factor (TF) in blood cells or surrounding tissue. During chemotherapy for lymphoma, the white blood cell count was significantly reduced at days 1 and 3, but significantly increased at days 7 and 9. At day 7 of chemotherapy, leukocyte TF mRNA levels were significantly increased. Plasma concentrations of granulocyte elastase derived-XDP (GEXDP) levels correlated with D-dimer levels, suggesting that almost all elevated D-dimer is GE-XDP. C-reactive protein (CRP), GE-XDP and D-dimer were significantly elevated in patients with infection, DIC or acute respiratory distress syndrome (ARDS). Analysis of patients with DIC or ARDS revealed that TF mRNA correlated with D-dimer, and GE XDP correlated with leukocyte count, CRP and D-dimer, suggesting that inflammatory changes due to thrombosis may cause the activation of leukocytes during chemotherapy. PMID- 16244578 TI - Identification of novel binding partners for tuberous sclerosis complex 2 (TSC2) by yeast two-hybrid approach. AB - AIM: To identify novel tuberous sclerosis complex (TSC2) binding partners by yeast two-hybrid screening. METHODS: The yeast two-hybrid system DupLEX-A developed by OriGene Technologies and Mouse embryo and HeLa cells cDNA libraries were used in this study. The "bait" constructs, containing full-length and truncated form of TSC2 were prepared. The expression of all constructs in yeast was confirmed by immunoblotting with specific anti-LexA antibodies. The suitability of generated constructs for screening was tested in autoactivation and nuclear translocation assays. Screening of mouse embryo and HeLa cDNA libraries with selected baits was carried out according to manufacturer's recommendations. Positive clones were selected using double selection procedure and further confirmed in mating assay. Isolated cDNA clones were identified by automated DNA sequencing and database searching. RESULTS: Extensive screening of two cDNA libraries from mouse embryo and HeLa cells with TSC2 baits led to the isolation of 102 positives clones. The specificity of interaction between TSC2 and binding proteins of selected clones was confirmed by mating assay for 83 clones. Sequencing of these clones indicated that they encode already known and novel TSC2-binding partners. CONCLUSION: The isolation of several known TSC2 binding partners, such as several isoforms of 14-3-3, demonstrates the validity of generated bait constructs and screening conditions. In addition, we have found a number of novel interactors, which encode cytoskeletal proteins and signaling molecules, such as Ser/Thr phosphatases. PMID- 16244579 TI - The role of expression of the components of proteome in the formation of molecular profile of human ovarian carcinoma A2780 cells sensitive and resistant to cisplatin. AB - AIM: To study the expression of proteins that characterize drug resistance, proliferation and apoptosis of human ovarian carcinoma cells. METHODS: The study was carried out on human ovarian carcinoma A2780 cells and on the A2780/DDP8 subline resistant to cisplatin. Expression of the surface and intracellular antigens (p53, Bcl-2, CD95, antigen of proliferating cells, metallothioneins, drug resistance proteins (P-glycoprotein (P-gp), glutathione-S-transferase), molecules of adhesion (E-cadherin, alpha- and beta-catenins) was studied by immunocytochemical method. RESULTS: It has been shown that the formation of the resistance to cisplatin in A2780/DDP8 cells is accompanied by the increase of expression of glutathione-S-transferase and Bcl-2, by the decrease of expression of CD95-antigene and proliferation potential of the cells, by appearance of EGF receptors and elevation of expression level of E-cadherin, alpha- and beta catenins, proving the enhancement of adhesive properties of tumor cells. CONCLUSION: Antiapoptotic program seems to be the leading mechanism of the development of the resistance to cisplatin in A2780/DDP8 cells and is realized via high expression of Bcl-2. During the development of drug resistance of A2780/DDP cells, the program of glutathione detoxification is functionally replacing the decrease of the content of metallothioneins. PMID- 16244580 TI - Expression of Bax, Bad and Bcl-2 proteins under x-radiation effect towards human breast carcinoma MCF-7 cells and their doxorubicin-resistant derivatives. AB - AIM: 1) To investigate the effect of X-rays on expression of Bax, Bad, and Bcl-2 proteins in human breast carcinoma MCF-7 cells, sensitive (MCF-7(wt)) and resistant (MCF-7(DOX/R)) to doxorubicin; 2) to measure X-radiation-induced DNA damage in order to clarify if changes in the expression of those proteins and DNA repair capacity can be responsible for the development of drug-resistance and for altered radio-sensitivity of MCF-7(DOX/R) cells. METHODS: Expression of Bax, Bad and Bcl-2 proteins was detected by Western-blot analysis. DNA damage induced by X radiation and the kinetics of subsequent DNA repair were estimated by alkaline DNA comet assay. RESULTS: We found an increase in the expression of pro-apoptotic Bax and Bad proteins and a decrease in the expression of Bcl-2 protein in MCF 7(wt) cells under the effect of X-radiation, whereas the expression of those proteins was not changed significantly in the irradiated MCF-7(DOX/R) cells. Different pattern of Bcl-2 expression was shown in MCF-7(DOX/R) cells compared with MCF-7(wt) cells. A decreased level of X-radiation-induced DNA damage and an increased DNA repair capacity were revealed in MCF-7(DOX/R) cells compared with MCF-7(wt) cells. CONCLUSIONS: The results of our study suggest that a reduced apoptotic response and an increased DNA repair capacity in MCF-7(DOX/R) cells might contribute to the resistance of these cells to X-radiation treatment. PMID- 16244581 TI - Hypoxia level and matrix metalloproteinases-2 and -9 activity in Lewis lung carcinoma: correlation with metastasis. AB - AIM: To investigate the relationship between tumor hypoxia in vivo, activity of matrix metalloproteinases (MMPs), and metastatic potential of tumor. MATERIALS AND METHODS: Lewis lung carcinoma (3LL) was used in this study. Total activity of MMP-2 and -9 in tumor was measured biochemically, tumor hypoxia level was assessed by (31)P NMR spectroscopy in tissue perchloric extracts. RESULTS: It was determined that hypoxia level in primary tumor has been concomitantly increasing along with tumor growth and correlated with metastasis level in lung. The positive correlation between hypoxia level and activities of MMP-2 and MMP-9 in primary tumor was registered. Moreover, the activity of MMP-2 and -9 in 3LL (primary tumor) directly correlates with metastasis level in lung. CONCLUSION: This study demonstrated that the growth of primary tumor is distinctly accompanied by an increase of tumor hypoxia level which positively correlates both with the activity of MMP-2 and -9 in primary tumor and metastatic efficiency. PMID- 16244582 TI - Response of the thymus and spleen of CBA mice with sarcoma 37 on intravenous and subcutaneous administration of syngeneic dendritic cells. AB - OBJECTIVE: To compare the effect of intravenous (i/v) and subcutaneous (s/c) infusion of syngeneic dendritic cells (DC) loaded or unloaded with lyzate of tumor cells (LTC) on apoptosis and proliferation of thymic and splenic cells in CBA mice with sarcoma 37 (S 37). METHODS: Experiments were carried out on CBA mice bearing transplanted ascitic form of S 37. DC loaded and unloaded with LTC were infused i/v and s/c (2 x 10(5) per animal) every 4 days (3 injections in total). Apoptosis and proliferation of thymic and splenic cells were evaluated using flow cytofluorimetry. RESULTS: It has been found that tumor development is accompanied by altered apoptosis and proliferation level in thymus and spleen. I/v infusion of syngeneic DC loaded and unloaded with LTC rendered the normalizing effect on the state of thymus in spleen in tumor-bearing animals. In the case of s/c infusion such effect of DC is less expressed. The use of DC, loaded with LTC irrespectively of the way of administration renders more durable effect on the functional state of these cells than the use of unloaded DC. CONCLUSION: Injection of syngeneic DC both loaded or unloaded with LTC renders a normalizing effect on the state of organs of immune system in tumor-bearing CBA mice. PMID- 16244583 TI - Galectin-3 and proliferating cell nuclear antigen (PCNA) expression in papillary thyroid carcinoma. AB - BACKGROUND AND AIM: To examine the relationship between galectin-3 and cell proliferation in thyroid tumor tissue. Galectin-3, a beta-galactoside binding protein, has recently been recognized as a promising molecular marker of thyroid malignancy, due to its high expression in thyroid carcinomas and absence from normal or benign thyroid tissue. However, its exact role in thyroid tumor biology is still unknown. PATIENTS AND METHODS: We examined the relationship between galectin-3 and cell proliferation by comparative immunostaining for galectin-3 and proliferating cell nuclear antigen (PCNA) in paraffin-embedded tissues from 126 cases of papillary thyroid carcinoma. RESULTS: Positive cytoplasmic immunostaining for galectin-3 was found in 115 (91.3%) cases. Nuclear staining for PCNA was detectable in 93 (74.4%) cases. A low level of PCNA staining (less than 10% positive cells) was found in 36 (28.6%) cases, moderate staining for PCNA (more than 10% but less than 30% positive cells) in 35 cases (27.8%), while highly increased PCNA expression (more than 30% positive cells) was found in 32 (25.4%) cases. Moderate or strong galectin-3 expression, found in 99 cases, was associated with highly increased PCNA staining in 28.3% of them but with no detectable PCNA expression in 24.3% of them. CONCLUSION: These results suggest that overexpression of galectin-3 is not clearly related to proliferative activity of papillary thyroid carcinoma cells as assessed by PCNA immunostaining. PMID- 16244584 TI - Effects of N-acylethanolamines and various antimitotic agents on apoptotic DNA fragmentation in conventionally normal and tumor tissue of human adrenals. AB - AIM: To study effects of N-acylethanolamines (NAE) and various antimitotic agents: taxol, colchicine, and cytochalasin B on the DNA fragmentation extent in conventionally normal (CNT) and tumor tissue of human adrenal cortex. METHODS: Six types of adrenal tumor tissue of 84 patients were analyzed. Slices of tissue was incubated at 37 degrees C for 3 h in the presence of NAEs and antimitotics, DNA was extracted and analysed in agarose gel. RESULTS: It was established that NAEs enhanced apoptosis in conventionally normal and tumor tissue of adrenal glands. In general, tumor tissue was more sensitive to NAEs and antimitotic compounds than conventionally normal tissue. NAEs in combination with colchicine and cytochalasin B enhanced DNA fragmentation in some types of tumor tissue and did not influence, or even reduced it in CNT. Taxol revealed selective action depending on tumor tissue type. Considerable individual differences were reported in sensitiveness of different types of tumors to the NAEs and antimitotic agents. CONCLUSIONS: Taxol and combination of NAEs with colchicine and cytochalasin B are inductors of apoptosis in the adrenal tumor cells and thus promising for further investigation. PMID- 16244585 TI - RAR-beta(1) overexpression in chromophobe renal cell carcinoma: a novel target for therapeutic intervention? AB - AIM: Retinoic acid (RA) has proven to possess modest but distinct activity in metastatic renal cell carcinoma (RCC), at least in a subgroup of patients. However, the exact molecular mechanisms leading to success or failure of RA application in individual patients are still unknown. As earlier studies have indicated that in RCC the RA receptor (RAR) beta might play a central role in RA signaling, we investigated the expression of the isoforms RAR-beta(1+2) in primary conventional and chromophobe RCC. METHODS: We used quantitative RT-PCR methodology to study RAR-beta(1) and RAR-beta(2) expression in ten primary conventional RCC samples (clear cell type), in two chromophobe RCC specimens, and the respective corresponding normal kidney tissues. The housekeeping genes RPS9 and RPLP0 were applied to normalize differences in mRNA quality and quantity. RESULTS: In contrast to conventional RCC samples, RAR-beta(1) was significantly overexpressed in both chromophobe tumors compared to the adjacent normal kidney tissue (p=0.03). On the contrary, RAR-beta(2) expression did neither differ significantly between conventional and chromophobe RCC (p=0.91) nor between malignant and normal kidney tissue (p>or=0.47). CONCLUSION: We demonstrate for the first time a significant and specific overexpression of RAR-beta(1) in chromophobe RCC. In future we will have to confirm this result within a larger number of samples. PMID- 16244586 TI - Vascular endothelial growth factor, c-erbB-2 and c-erbB-3 expression in colorectal adenoma and adenocarcinoma. AB - AIM: To analyze vascular endothelial growth factor (VEGF), c-erbB-2 and c-erbB-3 expression and to evaluate their relation to clinicopathologic parameters and pathogenesis of colorectal carcinoma. METHODS: Sections of adenoma, intramucosal carcinoma and adenocarcinoma were evaluated by immunohistochemistry in 85 malignant and 37 benign colorectal neoplasms for the expression of VEGF, c-erbB-2 and c-erbB-3 considering clinicopathological variables. RESULTS: VEGF was detected in comparable percentages of all neoplasm types while c-erbB-2 expression was detectable more frequently in adenoma than adenocarcinoma cases (65% vs 43%). Except for the correlation of c-erbB-3 expression with Dukes' staging, there was no correlation between the studied markers and grade of differentiation, Dukes' stage and localization of colorectal adenocarcinoma. c erbB-3 expression was seen more frequently in tubular adenomas, while c-erbB-2 expression was higher in tubulovillous and villous types. These differences were not statistically significant. The presence of distant metastasis and angiolymphatic invasion were identified as independent predictors of survival. A positive correlation was found between VEGF expression and lymphatic vessel invasion and regional lymph node involvement. CONCLUSION: These results suggest that VEGF, c-erbB-2, c-erbB-3 expression does not have prognostic value in colorectal cancer. VEGF expression may be implicated in the lymph node metastasis. PMID- 16244587 TI - Novel cytogenetic findings revealed by conventional cytogenetic and FISH analyses in leukaemia patients. AB - AIM: To describe novel cytogenetic findings in four leukaemia patients. METHODS: Conventional cytogenetic (CC) and fluorescence in situ hybridization (FISH) analyses were performed on bone marrow samples of four leukaemia patients. RESULTS: In this study, t(3;10)(q11;q25) and t(2;22)(p21;q11.2) were detected as novel translocations. t(8;16;21)(q22.1;q13;q22) and t(1;6;9;22)(p36.1;p21.3;q34;q11) were found as variant translocations, and these variant translocations were confirmed by Interphase-FISH and Multi-colour-FISH. CONCLUSION: Newly identified cytogenetic findings can lead us to characterize cytogenetic evolution of the haematological malignancies. Further investigations are certainly warranted to resolve the prognostic impact of these new cytogenetic abnormalities. PMID- 16244588 TI - Activity of cathepsin D and alpha(1)-antitrypsin in the blood serum of patients with mammary carcinoma. AB - THE AIM of this study was to determine the activity of cathepsin D and alpha(1) antitrypsin in the blood serum of patients with mammary carcinoma. PATIENTS AND METHODS: The study was conducted on 52 women operated for a unilateral breast tumor, divided into two groups, according to the number of metastases and tumor size. Cathepsin D activity was determined using the method of Anson, while alpha(1)-antitrypsin activity was determined according to the Eriksson method. RESULTS: Both groups of patients with mammary carcinoma were found to have higher activity of cathepsin D before the treatment compared to healthy females. After the surgery the enzyme activity increased significantly, whereas 6 months after the surgery it generally decreased. The activity of alpha(1)-antitrypsin was significantly lower in patients before the treatment than in the controls, while after 6 months an increase in alpha(1)-antitrypsin activity was observed. The correlation between activity of cathepsin D and alpha(1)-antitrypsin was revealed. High enzyme activity and low alpha(1)-antitrypsin activity may result from the stage of neoplastic transformation. CONCLUSION: The determination of cathepsin D activity together with alpha(1)-antitrypsin activity may serve as useful biochemical marker in monitoring of malignant changes in breast tumor. PMID- 16244589 TI - Global quality of life in patients who have undergone the hematopoietic stem cell transplantation: finding from transversal and retrospective study. AB - AIM: To analyse the factors which influence global quality of life in patients after the hematopoietic stem cell transplantation at the Department of Clinical Hematology of the Second Internal Clinic of the University Hospital and Medical Faculty of Charles University in Hradec Kralove, Czech Republic. PATIENTS AND METHODS: The total number of respondents after the transplantation from 2001 to 2003 was 95 and the return rate of questionnaires was 72.1% (71 respondents - 39 men and 32 women, average age - 55.5 years). The Czech version of an international generic European Quality of Life Questionnaire - Version EQ-5D was used. The influence of monitored factors (type of transplantation - autologous, allogenous, age, sex, education, poly-morbidity, marital status, religion and the time lapse from the hematopoietic stem cell transplantation) on the global quality of life in patients was determined by means of dispersion analysis. RESULTS: The above-mentioned factors proved statistically significant dependence of EQ-5D index and EQ-5 VAS on age (in both cases p<0.01), poly-morbidity (in both cases p<0.01) and on religion (in both cases p<0.01). EQ-5D index (dimensions of quality of life) and EQ-5D VAS (a subjective health condition) significantly decrease with increasing age and with a higher number of associated diseases, and are significantly higher in patients who believe in God compared to patient without religious beliefs. The influence of other factors on EQ-5D index and EQ-5D VAS was not proven as statistically significant. CONCLUSION: The global quality of life in patients who underwent the hematopoietic stem cell transplantation at the Department of Clinical Hematology of the Second Internal Clinic of the University Hospital and Medical Faculty of Charles University in Hradec Kralove, Czech Republic, is very high, which is seen from EQ-5D index (72.5%) and EQ-5D VAS (76.5%) values. PMID- 16244590 TI - DIC and lymphoma: editorial comment. PMID- 16244592 TI - What I think is... PMID- 16244593 TI - Harsh advice? PMID- 16244591 TI - Antiangiogenic activity of BAI1 in vivo: implications for gene therapy of human glioblastomas. AB - Glioblastomas are the most common primary brain tumors in adults. These tumors exhibit a high degree of vascularization, and malignant progression from astrocytoma to glioblastoma is often accompanied by increased angiogenesis and the upregulation of vascular endothelial growth factor and its receptors. In this study, we investigated the in vivo antiangiogenic and antitumor effects of brain specific angiogenesis inhibitor 1 (BAI1) using human glioblastoma cell lines. Glioblastoma cells were transduced with an adenoviral vector encoding BAI1 (AdBAI1), and Northern and Western blot analyses, respectively, demonstrated BAI1 mRNA and protein expression in the transduced tumor cells. Using an in vivo neovascularization assay, we found that angiogenesis surrounding AdBAI1 transduced glioblastoma cells transplanted into transparent skinfold chambers of SCID mice was significantly impaired compared to control treated cells. Additionally, in vivo inoculation with AdBAI1 of established subcutaneous or intracerebral transplanted tumors significantly impaired tumor growth and promoted increased mouse survival. Morphologically, the tumors exhibited signs of impaired angiogenesis, such as extensive necrosis and reduced intratumoral vascular density. Taken together, these data strongly indicate that BAI1 may be an excellent gene therapy candidate for the treatment of brain tumors, especially human glioblastomas. PMID- 16244594 TI - Double standard? PMID- 16244595 TI - Uncertainty and delay. PMID- 16244596 TI - Patient individuality. PMID- 16244597 TI - SHO pay scales. PMID- 16244598 TI - Must-have kit. PMID- 16244599 TI - Sleepless in surbiton. PMID- 16244600 TI - Ozone revolution? Or emperor's new clothes? PMID- 16244616 TI - Oral medicine--update for the dental practitioner: oral malodour. AB - This series provides an overview of current thinking in the more relevant areas of oral medicine for primary care practitioners, written by the authors while they were holding the Presidencies of the European Association for Oral Medicine and the British Society for Oral Medicine, respectively. A book containing additional material will be published. The series gives the detail necessary to assist the primary dental clinical team caring for patients with oral complaints that may be seen in general dental practice. Space precludes inclusion of illustrations of uncommon or rare disorders, or discussion of disorders affecting the hard tissues. Approaching the subject mainly by the symptomatic approach--as it largely relates to the presenting complaint--was considered to be a more helpful approach for GDPs rather than taking a diagnostic category approach. The clinical aspects of the relevant disorders are discussed, including a brief overview of the aetiology, detail on the clinical features and how the diagnosis is made. Guidance on management and when to refer is also provided, along with relevant websites which offer further detail. PMID- 16244615 TI - Science versus empiricism. AB - Few debates within dentistry have aroused such strong feelings as the issue of orthodontic growth guidance. Most orthodontists consider that appliances have little influence beyond the teeth and alveolus. If there is not room, it is argued, extractions are inevitable. Despite this a sizable minority, mostly of general dentists, believe that it is possible to influence skeletal form significantly. Why is there such lack of consensus? PMID- 16244617 TI - The management of the Muslim dental patient. AB - There are aspects of the practice of the religion of Islam that have some relevance to receiving dental treatment. This article aims to provide dentists with background knowledge of normal practices which may affect the treatment offered. The author does not attempt to inform the reader about Islam, but to assist the dentist in the management of a Muslim patient. Much of the content of this article describes how to manage a patient who is fasting during the Islamic month of Ramadan. Ramadan takes place this year in early October, lasting for 29 or 30 days. During Ramadan patients may present to dentists with the signs and symptoms described in this article. PMID- 16244618 TI - Dental and oral complications of lip and tongue piercings. AB - Piercing of the tongue and perioral regions is an increasingly popular expression of body art, with more patients coming in for a routine check-up with tongue and/or lip piercings. Several complications of oral piercing have been reported, some of which are life-threatening. In the present clinical survey the prevalence of both tongue and lip piercing complications in oral health was assessed in a group of 50 patients. The most common dental problem registered was chipping of the teeth, especially in association with tongue piercing. Gingival recession was seen as a result of lip piercing with studs. Post-procedural complications included oedema, haemorrhage and infection. Therefore, dentists and oral and maxillofacial surgeons should be given more authority to advise patients with oral and facial piercings or those who plan to acquire this type of body art. PMID- 16244625 TI - Insufficient incentives. PMID- 16244626 TI - Conflicting viewpoints. PMID- 16244627 TI - The dental practitioner and child protection in Scotland. AB - OBJECTIVES: To identify from general dental practitioners: undergraduate and postgraduate training experience in child protection; numbers of suspected cases of child physical abuse; reasons for failing to report suspicious cases of child physical abuse; knowledge of local child protection protocols and procedures for referral. MATERIALS AND METHODS: Postal questionnaires were sent to 500 randomly selected general dental practitioners in Scotland, with a further 200 sent to a random sample of the original 500 to increase response rate. RESULTS: Sixty-one per cent (306) of the original 500 questionnaires, and 35% (69) of the second random mail shot of 200 questionnaires were returned. Only 19% could remember any undergraduate training and 16% had been to a postgraduate lecture or seminar in child protection. Twenty-nine per cent of dentists had seen at least one suspicious case in their career. Only 8% of suspicious cases were referred on to the appropriate authorities. Reasons for failure to refer revealed that 11% were concerned about a negative impact on their practice, 34% feared family violence towards the child, 31% feared violence directed against them, and 48% feared litigation. Only 10% of dentists had been sent a copy of the local child protection guidelines on commencing work and only 15% had seen their Area Child Protection Committee (ACPC) Guidelines via any route. CONCLUSIONS: Due to lack of training or clear guidelines for dentists in Scotland, most practitioners were unsure what to do in the event of a suspicion of child abuse. Twenty-one per cent of dentists had encountered suspicious cases but failed to take any action. Dentists overwhelmingly requested appropriate training. This training should address dental competence in assessment of suspicious indicators and involve dentists in inter-agency child protection training. PMID- 16244628 TI - A study of visual and blood contamination on reprocessed endodontic files from general dental practice. AB - OBJECTIVE: This study examined methods used for reprocessing endodontic instruments in general dental practice and determined the degree of residual visual contamination and blood contamination on 250 reprocessed files collected from 25 general dental practices. MATERIALS AND METHODS: A questionnaire was administered to 25 general dental practitioners to obtain information on the re processing of used endodontic files. Ten files which had been used and reprocessed were also collected from each practice. These were examined visually under a dissecting light microscope for residual contamination and then tested for blood deposits using the Kastle-Meyer test. RESULTS: Nineteen of the 25 practices used stainless steel hand files. No practitioners used endodontic files as single use devices. Ninety-two per cent of the practitioners discarded and replaced files when they were bent or damaged. Several decontamination methods were reported. The two combinations employed most frequently were manual cleaning and autoclaving or manual cleaning, followed by ultrasonic cleaning and autoclaving. Of the 250 files, 75% showed some degree of visual contamination and seven percent tested positive for residual blood. Blood contaminated files were significantly more heavily contaminated when examined visually. Large variations were found in residual contamination of files collected from practices using the same methods of decontamination. CONCLUSIONS: While all practitioners re-used endodontic files, the variations in decontamination methods reported indicate a lack of clarity on best practice. This study demonstrates that endodontic files are not reliably decontaminated by methods currently employed in dental practice. PMID- 16244639 TI - 125 years of developments in dentistry, 1880-2005. Part 3: dental equipment and materials. PMID- 16244640 TI - Visual loss precluding acute onset of Budd-Chiari syndrome. PMID- 16244641 TI - Severe inflammation following iris fixated anterior chamber phakic intraocular lens for myopia. PMID- 16244642 TI - Neoadjuvant topical mitomycin C chemotherapy for conjunctival and corneal intraepithelial neoplasia. PMID- 16244643 TI - Ophthalmomyiasis externa caused by Dermatobia hominis: a successful treatment with oral ivermectin. PMID- 16244644 TI - Kingella kingae orbital cellulitis in a 3-year-old. PMID- 16244645 TI - The natural history of Vigabatrin-associated visual field defects in patients electing to continue their medication. PMID- 16244646 TI - Transpupillary thermotherapy for subfoveal choroidal neovascular membrane in choroidal osteoma. PMID- 16244647 TI - Pars plana vitrectomy for diabetic macular oedema: OCT and functional correlations. AB - PURPOSE: A prospective study to evaluate the macular structural and functional effects of pars plana vitrectomy (PPV) for persistent diffuse clinically significant macular oedema (CSMO). METHOD: A total of 12 patients with persistent diffuse CSMO were recruited and underwent assessment including best-corrected visual acuity, fundus fluorescein angiography, optical coherence tomography (OCT) and fine matrix mapping (FMM) at baseline and over a period of a year poststandard three-port PPV. RESULTS: The median baseline ETDRS letters score for all 12 patients was 52 (range 41-63) while at 12 months it had increased to 65 (range of 27-68), an improvement of two complete ETDRS lines (P=0.037). Similarly, there was an improvement in the perifoveal cone thresholds (P=0.02). The foveal thickening for all 12 patients ranged from a median of 183 to 751 microm (normal range 126-180 microm) and the macular volume ranged from a median of 2.13 to 6.42 mm(3) (normal <1.66 mm(3)). After surgery, both the median foveal thickness (from 334 to 280 microm) and median macular volume (from 3.24 to 2.61 mm(3)) demonstrated decreases over 12 months (P=0.01). On baseline OCT, the patients fell into two anatomically distinct groups: Group 1 (n=4) had a dome shaped thickened macula with a partial posterior hyaloid separation and a significantly higher foveal thickness and macular volume than Group 2 (n=8) which had a diffuse low-elevation profile of the thickened macula (P=0.007). CONCLUSIONS: In this prospective study of PPV for persistent fovea-involving CSMO there was structural and functional improvement. PMID- 16244648 TI - Glyoxalase 1 and glutathione reductase 1 regulate anxiety in mice. AB - Anxiety and fear are normal emotional responses to threatening situations. In human anxiety disorders--such as panic disorder, obsessive-compulsive disorder, post-traumatic stress disorder, social phobia, specific phobias and generalized anxiety disorder--these responses are exaggerated. The molecular mechanisms involved in the regulation of normal and pathological anxiety are mostly unknown. However, the availability of different inbred strains of mice offers an excellent model system in which to study the genetics of certain behavioural phenotypes. Here we report, using a combination of behavioural analysis of six inbred mouse strains with quantitative gene expression profiling of several brain regions, the identification of 17 genes with expression patterns that correlate with anxiety like behavioural phenotypes. To determine if two of the genes, glyoxalase 1 and glutathione reductase 1, have a causal role in the genesis of anxiety, we performed genetic manipulation using lentivirus-mediated gene transfer. Local overexpression of these genes in the mouse brain resulted in increased anxiety like behaviour, while local inhibition of glyoxalase 1 expression by RNA interference decreased the anxiety-like behaviour. Both of these genes are involved in oxidative stress metabolism, linking this pathway with anxiety related behaviour. PMID- 16244649 TI - Eye-specific effects of binocular rivalry in the human lateral geniculate nucleus. AB - When dissimilar images are presented to the two eyes, they compete for perceptual dominance so that each image is visible in turn for a few seconds while the other is suppressed. Such binocular rivalry is associated with relative suppression of local, eye-based representations that can also be modulated by high-level influences such as perceptual grouping. However, it is currently unclear how early in visual processing the suppression of eye-based signals can occur. Here we use high-resolution functional magnetic resonance imaging (fMRI) in conjunction with a new binocular rivalry stimulus to show that signals recorded from the human lateral geniculate nucleus (LGN) exhibit eye-specific suppression during rivalry. Regions of the LGN that show strong eye-preference independently show strongly reduced activity during binocular rivalry when the stimulus presented in their preferred eye is perceptually suppressed. The human LGN is thus the earliest stage of visual processing that reflects eye-specific dominance and suppression. PMID- 16244650 TI - Inducing and expanding regulatory T cell populations by foreign antigen. AB - Evidence suggests that regulatory T cells expressing the transcription factor Foxp3 develop extrathymically and intrathymically. Mechanisms of extrathymic induction require further scrutiny, especially as proliferation and/or phenotypic changes of preexisting suppressor cells must be distinguished from true de novo generation. Here we report the conversion of truly naive CD4(+) T cells into suppressor cells expressing Foxp3 by targeting of peptide-agonist ligands to dendritic cells and by analysis of Foxp3 expression at the level of single cells. We show that conversion was achieved by minute antigen doses with suboptimal dendritic cell activation. The addition of transforming growth factor-beta or the absence of interleukin 2 production, which reduces proliferation, enhanced the conversion rate. In addition, regulatory T cell populations induced in subimmunogenic conditions could subsequently be expanded by delivery of antigen in immunogenic conditions. The extrathymic generation and proliferation of regulatory T cells may contribute to self-tolerance as well as the poor immunogenicity of tumors and may be exploited clinically to prevent or reverse unwanted immunity. PMID- 16244651 TI - Regulation of lung injury and repair by Toll-like receptors and hyaluronan. AB - Mechanisms that regulate inflammation and repair after acute lung injury are incompletely understood. The extracellular matrix glycosaminoglycan hyaluronan is produced after tissue injury and impaired clearance results in unremitting inflammation. Here we report that hyaluronan degradation products require MyD88 and both Toll-like receptor (TLR)4 and TLR2 in vitro and in vivo to initiate inflammatory responses in acute lung injury. Hyaluronan fragments isolated from serum of individuals with acute lung injury stimulated macrophage chemokine production in a TLR4- and TLR2-dependent manner. Myd88(-/-) and Tlr4(-/-)Tlr2(-/ ) mice showed impaired transepithelial migration of inflammatory cells but decreased survival and enhanced epithelial cell apoptosis after lung injury. Lung epithelial cell-specific overexpression of high-molecular-mass hyaluronan was protective against acute lung injury. Furthermore, epithelial cell-surface hyaluronan was protective against apoptosis, in part, through TLR-dependent basal activation of NF-kappaB. Hyaluronan-TLR2 and hyaluronan-TLR4 interactions provide signals that initiate inflammatory responses, maintain epithelial cell integrity and promote recovery from acute lung injury. PMID- 16244652 TI - Sonic hedgehog myocardial gene therapy: tissue repair through transient reconstitution of embryonic signaling. AB - Sonic hedgehog (Shh) is a crucial regulator of organ development during embryogenesis. We investigated whether intramyocardial gene transfer of naked DNA encoding human Shh (phShh) could promote a favorable effect on recovery from acute and chronic myocardial ischemia in adult animals, not only by promoting neovascularization, but by broader effects, consistent with the role of this morphogen in embryogenesis. After Shh gene transfer, the hedgehog pathway was upregulated in mammalian fibroblasts and cardiomyocytes. This resulted in preservation of left ventricular function in both acute and chronic myocardial ischemia by enhanced neovascularization, and reduced fibrosis and cardiac apoptosis. Shh gene transfer also enhanced the contribution of bone marrow derived endothelial progenitor cells to myocardial neovascularization. These data suggest that Shh gene therapy may have considerable therapeutic potential in individuals with acute and chronic myocardial ischemia by triggering expression of multiple trophic factors and engendering tissue repair in the adult heart. PMID- 16244653 TI - Efficiency and power in genetic association studies. AB - We investigated selection and analysis of tag SNPs for genome-wide association studies by specifically examining the relationship between investment in genotyping and statistical power. Do pairwise or multimarker methods maximize efficiency and power? To what extent is power compromised when tags are selected from an incomplete resource such as HapMap? We addressed these questions using genotype data from the HapMap ENCODE project, association studies simulated under a realistic disease model, and empirical correction for multiple hypothesis testing. We demonstrate a haplotype-based tagging method that uniformly outperforms single-marker tests and methods for prioritization that markedly increase tagging efficiency. Examining all observed haplotypes for association, rather than just those that are proxies for known SNPs, increases power to detect rare causal alleles, at the cost of reduced power to detect common causal alleles. Power is robust to the completeness of the reference panel from which tags are selected. These findings have implications for prioritizing tag SNPs and interpreting association studies. PMID- 16244654 TI - Global hypomethylation of the genome in XX embryonic stem cells. AB - Embryonic stem (ES) cells are important tools in the study of gene function and may also become important in cell therapy applications. Establishment of stable XX ES cell lines from mouse blastocysts is relatively problematic owing to frequent loss of one of the two X chromosomes. Here we show that DNA methylation is globally reduced in XX ES cell lines and that this is attributable to the presence of two active X chromosomes. Hypomethylation affects both repetitive and unique sequences, the latter including differentially methylated regions that regulate expression of parentally imprinted genes. Methylation of differentially methylated regions can be restored coincident with elimination of an X chromosome in early-passage parthenogenetic ES cells, suggesting that selection against loss of methylation may provide the basis for X-chromosome instability. Finally, we show that hypomethylation is associated with reduced levels of the de novo DNA methyltransferases Dnmt3a and Dnmt3b and that ectopic expression of these factors restores global methylation levels. PMID- 16244655 TI - Mutation of Vps54 causes motor neuron disease and defective spermiogenesis in the wobbler mouse. AB - Vacuolar-vesicular protein sorting (Vps) factors are involved in vesicular trafficking in eukaryotic cells. We identified the missense mutation L967Q in Vps54 in the wobbler mouse, an animal model of amyotrophic lateral sclerosis, and also characterized a lethal allele, Vps54(beta-geo). Motoneuron survival and spermiogenesis are severely compromised in the wobbler mouse, indicating that Vps54 has an essential role in these processes. PMID- 16244656 TI - Cell-specific targeting of nanoparticles by multivalent attachment of small molecules. AB - Nanomaterials with precise biological functions have considerable potential for use in biomedical applications. Here we investigate whether multivalent attachment of small molecules can increase specific binding affinity and reveal new biological properties of such nanomaterials. We describe the parallel synthesis of a library comprising 146 nanoparticles decorated with different synthetic small molecules. Using fluorescent magnetic nanoparticles, we rapidly screened the library against different cell lines and discovered a series of nanoparticles with high specificity for endothelial cells, activated human macrophages or pancreatic cancer cells. Hits from the last-mentioned screen were shown to target pancreatic cancer in vivo. The method and described materials could facilitate development of functional nanomaterials for applications such as differentiating cell lines, detecting distinct cellular states and targeting specific cell types. PMID- 16244657 TI - Computational design and experimental validation of oligonucleotide-sensing allosteric ribozymes. AB - Allosteric RNAs operate as molecular switches that alter folding and function in response to ligand binding. A common type of natural allosteric RNAs is the riboswitch; designer RNAs with similar properties can be created by RNA engineering. We describe a computational approach for designing allosteric ribozymes triggered by binding oligonucleotides. Four universal types of RNA switches possessing AND, OR, YES and NOT Boolean logic functions were created in modular form, which allows ligand specificity to be changed without altering the catalytic core of the ribozyme. All computationally designed allosteric ribozymes were synthesized and experimentally tested in vitro. Engineered ribozymes exhibit >1,000-fold activation, demonstrate precise ligand specificity and function in molecular circuits in which the self-cleavage product of one RNA triggers the action of a second. This engineering approach provides a rapid and inexpensive way to create allosteric RNAs for constructing complex molecular circuits, nucleic acid detection systems and gene control elements. PMID- 16244658 TI - Efficient in vivo gene expression by trans-splicing adeno-associated viral vectors. AB - Although adeno-associated virus (AAV)-mediated gene therapy has been hindered by the small viral packaging capacity of the vector, trans-splicing AAV vectors are able to package twice the size of the vector genome. Unfortunately, the efficiency of current trans-splicing vectors is very low. Here we show that rational design of the gene splitting site has a profound influence on trans splicing vector-mediated gene expression. Using mRNA accumulation as a guide, we generated a set of efficient trans-splicing vectors and achieved widespread expression of the 6-kb DeltaH2-R19 mini-dystrophin gene in skeletal muscle of mdx mice, a model for Duchenne muscular dystrophy. The dystrophic phenotype was ameliorated in both adult and aged mice. This demonstrates the use of trans splicing vectors to efficiently express a large therapeutic structural protein. This strategy should be applicable to other large therapeutic genes or large transcription regulatory elements. PMID- 16244659 TI - An Anopheles transgenic sexing strain for vector control. AB - Genetic manipulation of mosquito species that serve as vectors for human malaria is a prerequisite to the implementation of gene transfer technologies for the control of vector-borne diseases. Here we report on the development of transgenic sexing lines for the mosquito Anopheles stephensi, the principal vector of human malaria in Asia. Male mosquitoes, expressing enhanced green fluorescent protein (EGFP) under the control of the beta2-tubulin promoter, are identified by their fluorescent gonads in as early as their 3(rd) instar larval stage, and can be efficiently separated from females using both manual methods and automated sorting machines. Importantly, beta2-EGFP males are not impaired in their mating ability and viable fluorescent spermatozoa are also detected in spermathecae of wild-type females mated with transgenic males. The transgenic mosquito lines described here combine most of the features desired and required for a safe application of transgenic methodologies to malaria-control programs. PMID- 16244660 TI - ERj1p uses a universal ribosomal adaptor site to coordinate the 80S ribosome at the membrane. AB - Ribosomes translating secretory and membrane proteins are targeted to the endoplasmic reticulum membrane and attach to the protein-conducting channel and ribosome-associated membrane proteins (RAMPs). Recently, a new RAMP, ERj1p, has been identified that recruits BiP to ribosomes and regulates translational activity. Here we present the cryo-EM structure of a ribosome-ERj1p complex, revealing how ERj1p coordinates the ribosome at the membrane and how allosteric effects may mediate ERj1p's regulatory activity. PMID- 16244661 TI - HIV-2 genomic RNA contains a novel type of IRES located downstream of its initiation codon. AB - Eukaryotic translation initiation begins with assembly of a 48S ribosomal complex at the 5' cap structure or at an internal ribosomal entry segment (IRES). In both cases, ribosomal positioning at the AUG codon requires a 5' untranslated region upstream from the initiation site. Here, we report that translation of the genomic RNA of human immunodeficiency virus type 2 takes place by attachment of the 48S ribosomal preinitiation complex to the coding region, with no need for an upstream 5' untranslated RNA sequence. This unusual mechanism is mediated by an RNA sequence that has features of an IRES with the unique ability to recruit ribosomes upstream from its core domain. A combination of translation assays and structural studies reveal that sequences located 50 nucleotides downstream of the AUG codon are crucial for IRES activity. PMID- 16244662 TI - A single spacer nucleotide determines the specificities of two mRNA regulatory proteins. AB - Regulation of messenger RNA is crucial in many contexts, including development, memory and cell growth. The 3' untranslated region is a rich repository of regulatory elements that bind proteins and microRNAs. Here we focus on PUF proteins, an important family of mRNA regulatory proteins crucial in stem-cell proliferation, pattern formation and synaptic plasticity. We show that two Caenorhabditis elegans PUF proteins, FBF and PUF-8, differ in RNA-binding specificity. FBF requires the presence of a single 'extra' nucleotide in the middle of an eight-nucleotide site, whereas PUF-8 requires its absence. A discrete protein segment is responsible for the difference. We propose that a structural distortion in the central region of FBF imposes the requirement for the additional nucleotide and that this mode of PUF specificity may be common. We suggest that new specificities can be designed and selected using the PUF scaffold. PMID- 16244664 TI - ERj1p has a basic role in protein biogenesis at the endoplasmic reticulum. AB - ERj1p is a membrane protein of the endoplasmic reticulum (ER) that can recruit the ER lumenal chaperone BiP to translating ribosomes. ERj1p can also modulate protein synthesis at initiation and is predicted to be a membrane-tethered transcription factor. Here we attribute the various functions of ERj1p to distinct regions within its cytosolic domain. A highly positively charged nonapeptide within this domain is necessary and sufficient for binding to ribosomes. Binding of ERj1p to ribosomes involves the 28S ribosomal RNA and occurs at the tunnel exit. Additionally, ERj1p has a dual regulatory role in gene expression: ERj1p inhibits translation in the absence of BiP, and another charged oligopeptide within the cytosolic domain of ERj1p mediates binding of the nuclear import factor importin beta and import into the nucleus, thereby paving the way for subsequent action on genomic DNA. PMID- 16244663 TI - Sequential phosphorylation and multisite interactions characterize specific target recognition by the FHA domain of Ki67. AB - The forkhead-associated (FHA) domain of human Ki67 interacts with the human nucleolar protein hNIFK, recognizing a 44-residue fragment, hNIFK226-269, phosphorylated at Thr234. Here we show that high-affinity binding requires sequential phosphorylation by two kinases, CDK1 and GSK3, yielding pThr238, pThr234 and pSer230. We have determined the solution structure of Ki67FHA in complex with the triply phosphorylated peptide hNIFK226-269(3P), revealing not only local recognition of pThr234 but also the extension of the beta-sheet of the FHA domain by the addition of a beta-strand of hNIFK. The structure of an FHA domain in complex with a biologically relevant binding partner provides insights into ligand specificity and potentially links the cancer marker protein Ki67 to a signaling pathway associated with cell fate specification. PMID- 16244665 TI - Induced structure of a helical switch as a mechanism to regulate enzymatic activity. AB - Herpesviruses encode a protease that is activated by homodimerization at high enzyme concentrations during lytic replication. The homodimer contains two active sites, which are distal from the dimer interface. Assignment of backbone NMR resonances and engineering of a redox switch show that two helices position a loop containing catalytic residues within each active site. PMID- 16244666 TI - Tankyrase-1 polymerization of poly(ADP-ribose) is required for spindle structure and function. AB - Poly(ADP-ribose) (PAR) is a large, negatively charged post-translational modification that is produced by polymerization of NAD+ by PAR polymerases (PARPs). There are at least 18 PARPs in the human genome, several of which have functions that are unknown. PAR modifications are dynamic; PAR structure depends on the balance between synthesis and hydrolysis by PAR glycohydrolase2. We previously found that PAR is enriched in vertebrate somatic-cell mitotic spindles and demonstrated a requirement for PAR in the assembly of Xenopus egg extract spindles. Here, we knockdown all characterized PARPs using RNA interference (RNAi), and identify tankyrase-1 as the PARP that is required for mitosis. Tankyrase-1 localizes to mitotic spindle poles, to telomeres and to the Golgi apparatus. Tankyrase-1 RNAi was recently shown to result in mitotic arrest, with abnormal chromosome distributions and spindle morphology observed--data that is interpreted as evidence of post-anaphase arrest induced by failure of telomere separation6. We show that tankyrase-1 RNAi results in pre-anaphase arrest, with intact sister-chromatid cohesion. We also demonstrate a requirement for tankyrase 1 in the assembly of bipolar spindles, and identify the spindle-pole protein NuMA as a substrate for covalent modification by tankyrase-1. PMID- 16244667 TI - Phosphatidylinositol transfer protein-alpha in netrin-1-induced PLC signalling and neurite outgrowth. AB - Neurite extension is essential for wiring the nervous system during development. Although several factors are known to regulate neurite outgrowth, the underlying mechanisms remain unclear. Here, we provide evidence for a role of phosphatidylinositol transfer protein-alpha (PlTPalpha) in neurite extension in response to netrin-1, an extracellular guidance cue. PlTPalpha interacts with the netrin receptor DCC (deleted in colorectal cancer) and neogenin. Netrin-1 stimulates PlTPalpha binding to DCC and to phosphatidylinositol (5) phosphate [Pl(5)P], increases its lipid-transfer activity and elevates hydrolysis of phosphatidylinositol bisphosphate (PlP2). In addition, the stimulated PIP2 hydrolysis requires PlTPalpha. Furthermore, cortical explants of PlTPalpha mutant mice are defective in extending neurites in response to netrin-1. Commissural neurons from chicken embryos expressing a dominant-negative PlTPalpha mutant show reduced axon outgrowth. Morpholino-mediated knockdown of PlTPalpha expression in zebrafish embryos leads to dose-dependent defects in motor-neuron axons and reduced numbers of spinal-cord neurons. Taken together, these results identify a crucial role for PlTPalpha in netrin-1-induced neurite outgrowth, revealing a signalling mechanism for DCC/neogenin and PlTPalpha regulation. PMID- 16244668 TI - The Polo kinase Plk4 functions in centriole duplication. AB - The human Polo-like kinase 1 (PLK1) and its functional homologues that are present in other eukaryotes have multiple, crucial roles in meiotic and mitotic cell division. By contrast, the functions of other mammalian Polo family members remain largely unknown. Plk4 is the most structurally divergent Polo family member; it is maximally expressed in actively dividing tissues and is essential for mouse embryonic development. Here, we identify Plk4 as a key regulator of centriole duplication. Both gain- and loss-of-function experiments demonstrate that Plk4 is required--in cooperation with Cdk2, CP110 and Hs-SAS6--for the precise reproduction of centrosomes during the cell cycle. These findings provide an attractive explanation for the crucial function of Plk4 in cell proliferation and have implications for the role of Polo kinases in tumorigenesis. PMID- 16244669 TI - Root gravitropism requires lateral root cap and epidermal cells for transport and response to a mobile auxin signal. AB - Re-orientation of Arabidopsis seedlings induces a rapid, asymmetric release of the growth regulator auxin from gravity-sensing columella cells at the root apex. The resulting lateral auxin gradient is hypothesized to drive differential cell expansion in elongation-zone tissues. We mapped those root tissues that function to transport or respond to auxin during a gravitropic response. Targeted expression of the auxin influx facilitator AUX1 demonstrated that root gravitropism requires auxin to be transported via the lateral root cap to all elongating epidermal cells. A three-dimensional model of the root elongation zone predicted that AUX1 causes the majority of auxin to accumulate in the epidermis. Selectively disrupting the auxin responsiveness of expanding epidermal cells by expressing a mutant form of the AUX/IAA17 protein, axr3-1, abolished root gravitropism. We conclude that gravitropic curvature in Arabidopsis roots is primarily driven by the differential expansion of epidermal cells in response to an influx-carrier-dependent auxin gradient. PMID- 16244672 TI - [Recently discovered virus--known symptoms]. PMID- 16244670 TI - The KLF4 tumour suppressor is a transcriptional repressor of p53 that acts as a context-dependent oncogene. AB - KLF4 (GKLF/EZF) encodes a transcription factor that is associated with both tumour suppression and oncogenesis. We describe the identification of KLF4 in a functional genomic screen for genes that bypass RAS(V12)-induced senescence. However, in untransformed cells, KLF4 acts as a potent inhibitor of proliferation. KLF4-induced arrest is bypassed by oncogenic RAS(V12) or by the RAS target cyclin-D1. Remarkably, inactivation of the cyclin-D1 target and the cell-cycle inhibitor p21CIP1 not only neutralizes the cytostatic action of KLF4, but also collaborates with KLF4 in oncogenic transformation. Conversely, KLF4 suppresses the expression of p53 by directly acting on its promoter, thereby allowing for RAS(V12)-mediated transformation and causing resistance to DNA damage-induced apoptosis. Consistently, KLF4 depletion from breast cancer cells restores p53 levels and causes p53-dependent apoptosis. These results unmask KLF4 as a regulator of p53 that oncogenically transforms cells as a function of p21CIP1 status. Furthermore, they provide a mechanistic explanation for the context-dependent oncogenic or tumour-suppressor functions of KLF4. PMID- 16244673 TI - [Reporting on suicide and health hazards]. PMID- 16244675 TI - [Can ear temperature measurement be used in a hospital?]. AB - BACKGROUND: There are two principle means of infrared thermometry: tympanic thermometry and ear canal thermometry. The purpose of this study was to examine the reliability of these ear thermometers compared with the digital rectal thermometer. MATERIAL AND METHODS: We used two different infrared ear thermometers in this study; one type measured the tympanic temperature, the other the temperature in the ear canal. The rectal temperature was defined as the gold standard. The temperatures measured with the ear thermometers were compared to rectal temperature on 213 patients older than 18 years at Haukeland University Hospital. RESULTS: The tympanic and ear canal measurements showed an average of 0.5 and 0.2 degrees Celsius respectively lower than the rectal temperature. The difference between the two types of ear thermometers was 0.4 degrees (p < 0.001, 95% CI 0.18 to 0.42). By using the tympanic thermometer, the sensitivity of detecting fever was 14% and the negative predictive value was 89%. Ear canal thermometry had 55% sensitivity for finding fever and a negative predictive value of 95%. INTERPRETATION: There are many advantages by using ear thermometers, but the accuracy and ability to detect fever is not good enough for it to be recommended in a clinical practice. We recommend using rectal thermometers. PMID- 16244674 TI - [Self-reported medical drug use among 15-16 year-old adolescents in Norway]. AB - BACKGROUND: Little information is available about Norwegian adolescents' use of medical drugs. MATERIAL AND METHODS: Self-reported drug use was recorded by means of a self-administered questionnaire handed out in classrooms to all 10th grade students in four counties. In all, 5846 boys and 5862 girls participated during the years 2000-2002. RESULTS: Forty-one percent of boys and 64% of girls had used drugs during the last four weeks. Non-prescription analgesics had been used by 25% of the boys and by more than half of the girls. Daily use of prescription analgesics was reported by approximately 1% in both sexes, while more girls than boys had used these drugs during the last four weeks. In all, 15% of the boys and 16% of the girls reported use of allergy medicines during the last four weeks and 6% of the boys and 7% of the girls had used asthma medicines. During the same period, 1.4% of the boys and 1.8% of the girls had used hypnotics. Anxiolytics had been used by 1.9%, while 0.8% reported use of antidepressants. INTERPRETATION: Drug use is common among adolescents in Norway. Except for analgesics, there were only minor sex differences in drug use. The proportion of drug users did not vary significantly between counties. PMID- 16244676 TI - [Human metapneumovirus--virologic and diagnostic aspects]. AB - Human metapneumovirus (hMPV) is a pathogenic respiratory virus, discovered in 2001. The virus is part of the family paramyxoviridae and is comparatively closely related to respiratory syncytial virus (RSV). The clinical manifestations of a hMPV infection are rather similar to infections caused by RSV, ranging from mild upper airway disease to severe pneumonia. Detection of hMPV RNA by polymerase chain reaction (PCR) is the only established routine diagnostic method. The virus is easily cultivated in several types of cell culture lines, however, cell pathology (cytopathic effect) is irregularly observed. Our department has developed an indirect immunofluorescence test based on polyvalent rabbit antibodies. This method appears to give reliable results, detecting hMPV both in cell culture and directly on patient samples. Nasopharyngeal aspirate is the preferred material for diagnostic purposes in small children. A nasopharyngeal swab, or alternatively a throat swab, can be used for older children and adults. Samples from lower airways are appropriate under special circumstances. Serology is not available for routine diagnostic use. PMID- 16244677 TI - [Human metapneumovirus--occurrence and clinical significance]. AB - BACKGROUND: Human metapneumovirus (hMPV) was isolated for the first time in 2001 from young children with acute respiratory tract infection. MATERIALS AND METHODS: Review of published data on the clinical impact of hMPV and our own experience with hMPV during three winter seasons. We present four cases of severe hMPV childhood infections. RESULTS: Human metapneumovirus accounts for approximately 10% of respiratory tract infections that are not related to previously known etiologic agents. The virus seems to be distributed worldwide and to have a seasonal distribution. During a short epidemic, 60% of our isolates from children with respiratory infection tested positive for hMPV. Serologic studies have shown that by the age of five, virtually all children have been exposed to the virus and reinfections appear to be common. Human metapneumovirus may cause mild respiratory tract infection. Small children, elderly and immunocompromised individuals are, however, at risk of severe disease and hospitalization. Although the clinical manifestations of hMPV resemble those of respiratory syncytial virus, we have observed that hMPV more often causes severe pneumonia in hospitalized children. INTERPRETATION: Human metapneumovirus is an important cause of acute respiratory tract infection in children. The virus may cause severe disease in patients at risk. We recommend identification and isolation of hospitalized hMPV-infected patients. PMID- 16244678 TI - [Phencyclidine--angel dust]. AB - BACKGROUND: For the first time in ten years, phencyclidine (PCP) has been confiscated in Norway. Physicians should be aware of this substance when treating intoxications. METHODS: Relevant literature was identified by search in Medline. This review presents the pharmacological properties and effects of phencyclidine as well as symptoms and treatment of phencyclidine intoxication. RESULTS AND INTERPRETATION: Phencyclidine exhibits hallucinogenic, depressant and stimulant properties. Phencyclidine interacts as an antagonist to the N-methyl-D-aspartate (NMDA) receptor in the central nervous system. Severe NMDA receptor hypofunction can elicit clinical symptoms similar to a schizophrenic episode. PMID- 16244679 TI - [Cardiac ion channel disorders--diagnosis and treatment]. AB - BACKGROUND: Inherited arrhythmogenic disorders are a group of genetically determined diseases characterised by ventricular tachyarrhythmias sometimes leading to sudden death. The molecular bases of these disorders are mutations in genes coding for various cardiac ion channels. The most common cardiac ion channel disease is the long QT syndrome. This syndrome is rare, but probably more common in Norway than previously expected. We have recently started genetic testing for cardiac ion channel disorders at Rikshospitalet University Hospital in Oslo. This review describes the current understanding of the etiology, prognosis and management of cardiac ion channel disorders, based on literature and our own clinical experience. INTERPRETATION: Cardiac ion channel disorders may lead to sudden cardiac death. Prophylactic and life-saving therapies are available for many of these disorders. Therapy and risk stratification depend on the clinical presentation, the ECG pattern, and which gene is mutated. Genetic testing offers the opportunity to exclude individual family members as mutation carriers. PMID- 16244680 TI - [DNA-based diagnostics of long QT syndrome]. AB - BACKGROUND: Long QT syndrome is characterised by inherited long QT interval on the ECG and increased risk for syncope and sudden death caused by arrhythmias. For Romano-Ward syndrome and Jervell and Lange-Nielsen syndrome DNA based diagnostics are available. MATERIALS AND METHODS: This paper is a summary of our experience with DNA-based diagnostics of LQTS since the autumn of 2003. The diagnostic analyses are performed by sequencing the exons of five genes, KCNQ1, HERG, SCN5A, minK and MiRP1. RESULTS AND INTERPRETATIONS: As of mid-January 2005, 56 probands with long QT syndrome have been referred for genetic testing. We have identified an underlying mutation in 64% of the patients. Mutations in the KCNQ1 gene are most frequent in Norwegian long QT syndrome patients, as 61% of the patients have their mutation in this gene. The detection of a mutation in the probands has led to genetic testing of 215 relatives; 99 out of these are heterozygous for the mutation present in the family. Heterozygous patients have been referred to a cardiologist. Of the 43 that have been referred to follow up at the department of cardiology at Rikshospitalet, 35 have started treatment with beta blockers to reduce the risk of arrhythmias. Thus, DNA-based diagnostics has clinical significance leading to prophylactic treatment of long QT syndrome patients. Compared to evaluation of ECG, which is negative in 30% of mutation carriers, the sensitivity of DNA-based diagnostics of relatives of probands with a known mutation, is close to 1. PMID- 16244681 TI - [Slipped capital femoral epiphysis treated with a specially designed screw]. AB - BACKGROUND: In 1992 we developed a specially designed Olmed screw for treatment of slipped capital femoral epiphysis. The segment of which the screw is threaded is shorter than a regular Olmed screw. In that way the threaded portion of the screw can be placed entirely within the epiphysis. This makes the shank of the screw smooth through the physis; the purpose of this is to allow continued growth in the neck of femur. The threads are reverse cutting to make the screw easier to remove. MATERIAL AND METHODS: From 1992 to 2004, we treated 18 hips with slipped capital femoral epiphysis; all patients were treated percutaneously with in situ fixation. We have retrospectively reviewed the medical records and radiographs. Nine patients were reviewed clinically and radiographically after an average follow up of seven year and nine months. RESULTS: There were no difficulties with the surgical procedure, including removal of the screw. No further slippage, avascular necrosis or chondrolysis occurred. Radiographs showed that the screw allows continued growth of the neck of femur. Most of the patients had decreased internal rotation, but few had symptoms at follow up. PMID- 16244682 TI - [Malignant hyperthermia--a hereditary and potentially life-threatening condition]. AB - BACKGROUND: Malignant hyperthermia is a rare and possibly life-threatening complication to anaesthesia. It manifests in susceptible individuals as a hypermetabolic response on exposure to halogenated anaesthetics and depolarising muscle relaxants. Susceptibility to malignant hyperthermia is inherited as an autosomal dominant disorder and is associated with myopathies. MATERIAL AND METHODS: We present a Norwegian family with central core disease and malignant hyperthermia susceptibility. A novel mutation (c.14558C>T) in the ryanodine receptor gene (RYR1), causing an amino acid change of a highly conserved residue (Thr4853Ile), has been identified in this family. We present a review of the literature on this disorder. RESULTS AND INTERPRETATION: Modern medical treatment, including the use of dantrolene, has significantly reduced the mortality of malignant hyperthermia. The identification of susceptibility to malignant hyperthermia in patients can possibly further reduce the risk of death during and after anaesthesia. A contracture test of muscular tissue is performed in patients with suspected malignant hyperthermia and should be considered in family members. Molecular genetic examinations might be considered in some cases. PMID- 16244683 TI - [Thoracoscopic sympathectomy in the treatment of palmar hyperhidrosis and facial blushing]. AB - BACKGROUND: Severe palmar hyperhidrosis and facial blushing are conditions connected with considerable psychosocial burden. Conservative treatment is often of limited value. Several reports have demonstrated high success rates in thoracoscopic sympathectomy. Treatment results from Norwegian centres have not previously been published. MATERIAL AND METHODS: The long term effects of thoracoscopic sympathectomy performed between 1998 and 2002 at Aker University Hospital were evaluated by questionnaire, using VAS scores (visual analogue scales) obtained from 72 out of 76 operated patients. RESULTS: In palmar hyperhidrosis, a reduction in VAS score above 3 was reported in 27 of 28 patients (from mean 9.5 +/- 0.2 to 0.8 + 0.4). In facial blushing, 39 of 44 patients reported a reduction in VAS score above 3 (from mean 8.9 +/- 0.3 to 2.5 +/- 0.4). The overall satisfaction rate was 90 %. A majority of patients (86 %) experienced variable but lasting compensatory general hyperhidrosis. However, only a few patients found this condition disabling to the extent that it caused regret (3 patients) or dissatisfaction (4 patients). CONCLUSION: Thoracoscopic sympathectomy markedly reduces palmar hyperhidrosis and facial blushing, but the indication should be strong and the information thorough because of compensatory hyperhidrosis. PMID- 16244684 TI - [Self-help programs in drug addiction therapy]. AB - BACKGROUND: Treatment of drug addiction is complex; hence there is a debate in the field of what is the best approach. An honest desire to stop using drugs is the entry ticket into the self-help programmes Alcoholics Anonymous (AA) and Narcotics Anonymous (NA); these groups are a possible supplement to ordinary treatment. They employ the twelve-step programme and are found in most large cities in Norway and around the world. The usefulness of these groups has been disputed, and few Norwegian treatment facilities have developed a systematic cooperation with them. The purpose of this study was to examine whether the use of self-help groups is associated with higher rates of abstinence two years after participants started attending groups. MATERIALS AND METHODS: 114 patients, 59 with alcohol addiction and 55 with drug addiction, who started in self-help groups after a drug treatment, were approached two years later with a questionnaire. The response rate was 66%; 6 (5%) patients were dead. RESULTS: Intention to treat analysis showed that 38% participated in self-help programmes two years after treatment. Of regular participants, 81% had been abstinent over the previous 6 months, compared with only 26% among non-participants. Logistic regression analysis showed odds ratio = 12.6, 95% CI (4.1-38.3), p < 0.001, for participation and abstinence. CONCLUSION: The study has several methodological problems; particularly that correlation does not necessarily indicate causality. These problems are discussed here; the conclusion is that the probability of a positive effect is strong enough to recommend participation in self-help groups as a supplement to drug addiction treatment. PMID- 16244685 TI - [Is there a difference between low-molecular-weight heparins?]. AB - Low-molecular-weight heparins share many properties and are commonly referred to as a group, but structurally and pharmacologically they are dissimilar. The size spectrum of the heparin molecules varies between the different products and as assessed in vitro, their anticoagulant properties differ. In particular, the ratio anti-factor Xa : anti-factor IIa activities varies. The clinical consequences of these differences are unknown. The efficacy and safety of two different low-molecular-weight heparins have been compared in only a few clinical studies; no significant differences in outcome were shown. However, low-molecular weight heparins should be used according to the approved indication for each product and in doses shown effective and safe in clinical studies. A change from one low-molecular-weight heparin to another in the same patient should be avoided. Fondaparinux is a synthetic penta-saccharide which may be regarded as an extreme low-molecular-weight heparin with a ratio of anti-factor Xa : anti-factor IIa activity as 1 : 0, and with a promising efficacy/safety profile. So far, the approved clinical indication for its use is limited to prophylaxis in orthopaedic surgery. PMID- 16244686 TI - [How can women with chronic muscular pain change their bodily habits?]. AB - BACKGROUND: Each person develops their own posture and patterns of movement; our bodies are marked by the life we live. Our ways of being imply that the body is used and strained in different ways, and these habits are decisive for what kind of functional problems we may develop. A group-based treatment programme for women with chronic muscular pain includes movement training and group discussions once a week over the course of ten months, aimed at changing straining bodily habits. The study explores what the participants appreciate as meaningful and beneficial effects of this programme. MATERIAL AND METHODS: A treatment group of ten women with long-lasting chronic muscular pain was organised at the start of the 21st century. Seven of them completed the programme. Transcriptions from individual interviews with the participants were analysed using a phenomenological analysis. RESULTS: The participants made new bodily experiences, changed their bodily habits and developed strategies for handling stress and pain in daily life. INTERPRETATION: Group treatment may be a beneficial offer in primary care to women with chronic muscular pain. PMID- 16244687 TI - [Pain as a health problem]. PMID- 16244689 TI - [About sugar, lactic acid and pain]. PMID- 16244688 TI - [Pain treatment in patients on chronic opioid therapy and in drug abusers]. AB - BACKGROUND: Substance abuse in pain management can undermine treatment compliance, the effectiveness of therapies, and social support and functioning. MATERIAL: An overview is given regarding pain management for patients on chronic use of opioids and for patients with drug dependence and substance abuse. RESULTS: Clinical experience as well as relevant documentation demonstrates that management of these patients remains one of the most challenging problems in clinical medicine. CONCLUSION: Assessment and treatment plans for this patient group require an understanding of tolerance, physical dependence and drug abuse. It is imperative that the patient's ongoing treatment is planned in a multidisciplinary setting, so as to assure a consistent, structured and shared approach by all caregivers. PMID- 16244691 TI - [Hospital discharge information as a communication tool]. AB - BACKGROUND: The main objective of the list patient health reform has been to improve the quality of local medical service. In European healthcare systems, there has been a need for coordination between primary and secondary care. It has been claimed that the communication between GPs and specialised health care is insufficient, particularly for patients with an extended need of care. MATERIAL AND METHODS: Data on collaboration and satisfaction with specialised health services was collected in a cross-sectional questionnaire survey comprising all Norwegian GPs in 2004 (N = 633); the response rate was 48%. RESULTS: Median delay in receiving patient information was one week or more for inpatient and outpatient treatment. Overall, GPs were satisfied with the hospitals' discharge reports, except for patients needing extended follow up by GPs after discharge. In these patients, GPs more frequently needed to contact the hospital because of insufficient data or unacceptable delays. Female GPs were less satisfied with information services in patients with increased needs. There were also geographical differences in satisfaction. INTERPRETATION: In patients with an increased need for follow up, GPs are less satisfied with hospital information service. PMID- 16244690 TI - [The list patient scheme and the growth insurance expenditure in general practice]. AB - BACKGROUND: Fee-for-item payments, mainly from the National Insurance Administration, are the main source of remuneration for primary care physicians in Norway. The aim of the present study was to describe the development in the National Insurance Administration's expenditure for primary physician services before and after the introduction of a list patient scheme in 2001. MATERIAL AND METHODS: The analyses were performed on data from the National Insurance Administration and data on the number of physician in relation to population from Statistics Norway. RESULTS: During the period 1998-2003, expenditure increased by 84%, from NOK 1.37 billion to NOK 2.52 billion. The main finding is that nearly all the increase can be explained by increases in the number of physicians, population size and the level of fees. INTERPRETATION: One interpretation of our findings is that to a large extent the National Insurance Administration's expenditure on primary physician services can be controlled by regulating the number of physicians and the level of fees. PMID- 16244692 TI - [Sense and sensibility in delivery care]. AB - The organisation of delivery care engages both professional and laypersons. When the Norwegian princess Martha Louise in 2005 chose to give birth at home instead of in hospital, the intensity of the debate in media increased. Some professionals claim with great persuasion that among selected low-risk women, birth at home or in small midwifery units is as safe as in larger delivery units in hospitals. The scientific evidence for this statement is, however, weak. The Norwegian National Advisory Committee for Maternal Care is an important contributor to this debate. The Committee should, however, pay more respect to the scientific disagreement within the field. PMID- 16244693 TI - [Coordination in psychiatry--a problem of the border between the fields?]. PMID- 16244695 TI - [Women's voices in the Northwest Russia]. PMID- 16244698 TI - [Radical treatment of prostatic cancer in Norway]. PMID- 16244700 TI - [Cryotherapy in prostatic cancer]. PMID- 16244702 TI - Glycan analysis of the chicken synaptic plasma membrane glycoproteins--a major synaptic N-glycan carries the LewisX determinant. AB - The majority of synaptic plasma membrane components are glycosylated. It is now widely accepted that this post-translational modification is crucial during the establishment, maintenance and function of the nervous system. Despite its significance, structural information about the glycosylation of nervous system specific glycoproteins is very limited. In the present study the major glycan structures of the chicken synaptic plasma membrane (SPM) associated glycoprotein glycans were determined. N-glycans were released by hydrazinolysis, labelled with 2-aminobenzamide, treated with neuraminidase and subsequently fractionated by size exclusion chromatography. Individual fractions were characterized by the combination of high-pressure liquid chromatography, exoglycosidase treatment or reagent array analysis method (RAAM). In addition to oligomannose-type glycans, core-fucosylated complex glycans with biantennary bisecting glycans carrying the LewisX epitope were most abundant. The overall chicken glycan profile was strikingly similar to the rat brain glycan profile. The presence of the LewisX determinant in relatively large proportions suggests a tissue-specific function for these glycans. PMID- 16244703 TI - Ber-H2 (CD30) immunohistochemical staining of human fetal tissues. AB - OBJECTIVE: CD30 antigen has long been considered to be restricted to the tumour cells of Hodgkin's disease and of anaplastic large cell lymphoma as well as to T and B activated lymphocytes. It is now apparent that the range of normal and neoplastic cells, which may express CD30 antigen, is much wider than was at first thought. In order to gain insight into the physiological function of CD30 antigen, we studied the distribution of its expression in the tissues of fetuses from week 8th to week 16th. MATERIALS AND METHODS: We investigated the immunohistochemical expression of CD30 antigen in paraffin-embedded tissue samples representing all systems from 30 fetuses after therapeutic abortion at 8th to 10th and 12th to 16th week of gestation, respectively, using the monoclonal antibody Ber-H2. RESULTS: Our results demonstrated that CD30 is expressed early in human fetal development (8th to 10th week of gestation) in several fetal tissues derived from all three germ layers (gastrointestinal tract, special glands of the postpharyngeal foregut, urinary, musculoskeletal, reproductive, nervous, endocrine systems), with the exception of the skin and hematolymphoid system (thymus), in which the antigen is expressed later on (10th week onwards). Expression of CD30 was restricted to the hematolymphoid system in the 12-16 weeks of gestation. No expression of the marker was observed in the respiratory and cardiovascular systems during the entire period examined. CONCLUSIONS: CD30 antigen is of importance in cell development, and proliferation. It is also pathway-related to terminal differentiation in many fetal tissues and organs. PMID- 16244704 TI - Structural evolution of the protein kinase-like superfamily. AB - The protein kinase family is large and important, but it is only one family in a larger superfamily of homologous kinases that phosphorylate a variety of substrates and play important roles in all three superkingdoms of life. We used a carefully constructed structural alignment of selected kinases as the basis for a study of the structural evolution of the protein kinase-like superfamily. The comparison of structures revealed a "universal core" domain consisting only of regions required for ATP binding and the phosphotransfer reaction. Remarkably, even within the universal core some kinase structures display notable changes, while still retaining essential activity. Hence, the protein kinase-like superfamily has undergone substantial structural and sequence revision over long evolutionary timescales. We constructed a phylogenetic tree for the superfamily using a novel approach that allowed for the combination of sequence and structure information into a unified quantitative analysis. When considered against the backdrop of species distribution and other metrics, our tree provides a compelling scenario for the development of the various kinase families from a shared common ancestor. We propose that most of the so-called "atypical kinases" are not intermittently derived from protein kinases, but rather diverged early in evolution to form a distinct phyletic group. Within the atypical kinases, the aminoglycoside and choline kinase families appear to share the closest relationship. These two families in turn appear to be the most closely related to the protein kinase family. In addition, our analysis suggests that the actin fragmin kinase, an atypical protein kinase, is more closely related to the phosphoinositide-3 kinase family than to the protein kinase family. The two most divergent families, alpha-kinases and phosphatidylinositol phosphate kinases (PIPKs), appear to have distinct evolutionary histories. While the PIPKs probably have an evolutionary relationship with the rest of the kinase superfamily, the relationship appears to be very distant (and perhaps indirect). Conversely, the alpha-kinases appear to be an exception to the scenario of early divergence for the atypical kinases: they apparently arose relatively recently in eukaryotes. We present possible scenarios for the derivation of the alpha-kinases from an extant kinase fold. PMID- 16244705 TI - Positive selection of Iris, a retroviral envelope-derived host gene in Drosophila melanogaster. AB - Eukaryotic genomes can usurp enzymatic functions encoded by mobile elements for their own use. A particularly interesting kind of acquisition involves the domestication of retroviral envelope genes, which confer infectious membrane fusion ability to retroviruses. So far, these examples have been limited to vertebrate genomes, including primates where the domesticated envelope is under purifying selection to assist placental function. Here, we show that in Drosophila genomes, a previously unannotated gene (CG4715, renamed Iris) was domesticated from a novel, active Kanga lineage of insect retroviruses at least 25 million years ago, and has since been maintained as a host gene that is expressed in all adult tissues. Iris and the envelope genes from Kanga retroviruses are homologous to those found in insect baculoviruses and gypsy and roo insect retroviruses. Two separate envelope domestications from the Kanga and roo retroviruses have taken place, in fruit fly and mosquito genomes, respectively. Whereas retroviral envelopes are proteolytically cleaved into the ligand-interaction and membrane-fusion domains, Iris appears to lack this cleavage site. In the takahashii/suzukii species groups of Drosophila, we find that Iris has tandemly duplicated to give rise to two genes (Iris-A and Iris-B). Iris-B has significantly diverged from the Iris-A lineage, primarily because of the "invention" of an intron de novo in what was previously exonic sequence. Unlike domesticated retroviral envelope genes in mammals, we find that Iris has been subject to strong positive selection between Drosophila species. The rapid, adaptive evolution of Iris is sufficient to unambiguously distinguish the phylogenies of three closely related sibling species of Drosophila (D. simulans, D. sechellia, and D. mauritiana), a discriminative power previously described only for a putative "speciation gene." Iris represents the first instance of a retroviral envelope-derived host gene outside vertebrates. It is also the first example of a retroviral envelope gene that has been found to be subject to positive selection following its domestication. The unusual selective pressures acting on Iris suggest that it is an active participant in an ongoing genetic conflict. We propose a model in which Iris has "switched sides," having been recruited by host genomes to combat baculoviruses and retroviruses, which employ homologous envelope genes to mediate infection. PMID- 16244707 TI - The evolutionary value of recombination is constrained by genome modularity. AB - Genetic recombination is a fundamental evolutionary mechanism promoting biological adaptation. Using engineered recombinants of the small single-stranded DNA plant virus, Maize streak virus (MSV), we experimentally demonstrate that fragments of genetic material only function optimally if they reside within genomes similar to those in which they evolved. The degree of similarity necessary for optimal functionality is correlated with the complexity of intragenomic interaction networks within which genome fragments must function. There is a striking correlation between our experimental results and the types of MSV recombinants that are detectable in nature, indicating that obligatory maintenance of intragenome interaction networks strongly constrains the evolutionary value of recombination for this virus and probably for genomes in general. PMID- 16244706 TI - Evolutionary comparison provides evidence for pathogenicity of RMRP mutations. AB - Cartilage-hair hypoplasia (CHH) is a pleiotropic disease caused by recessive mutations in the RMRP gene that result in a wide spectrum of manifestations including short stature, sparse hair, metaphyseal dysplasia, anemia, immune deficiency, and increased incidence of cancer. Molecular diagnosis of CHH has implications for management, prognosis, follow-up, and genetic counseling of affected patients and their families. We report 20 novel mutations in 36 patients with CHH and describe the associated phenotypic spectrum. Given the high mutational heterogeneity (62 mutations reported to date), the high frequency of variations in the region (eight single nucleotide polymorphisms in and around RMRP), and the fact that RMRP is not translated into protein, prediction of mutation pathogenicity is difficult. We addressed this issue by a comparative genomic approach and aligned the genomic sequences of RMRP gene in the entire class of mammals. We found that putative pathogenic mutations are located in highly conserved nucleotides, whereas polymorphisms are located in non-conserved positions. We conclude that the abundance of variations in this small gene is remarkable and at odds with its high conservation through species; it is unclear whether these variations are caused by a high local mutation rate, a failure of repair mechanisms, or a relaxed selective pressure. The marked diversity of mutations in RMRP and the low homozygosity rate in our patient population indicate that CHH is more common than previously estimated, but may go unrecognized because of its variable clinical presentation. Thus, RMRP molecular testing may be indicated in individuals with isolated metaphyseal dysplasia, anemia, or immune dysregulation. PMID- 16244708 TI - Identification and characterization of tyrosylprotein sulfotransferase from human saliva. AB - Tyrosylprotein sulfotransferase (TPST), the enzyme responsible for the sulfation of tyrosine residues, has been identified and characterized in submandibular salivary glands previously (William et al. Arch Biochem Biophys 338: 90-96). Tyrosylprotein sulfotransferase catalyses the sulfation of a variety of secretory and membrane proteins and is believed to be present only in the cell. In the present study, this enzyme was identified for the first time in human saliva. Analysis of human saliva and parotid saliva for the presence of tyrosylprotein sulfotransferase revealed tyrosine sulfating activity displayed by both whole saliva and parotid saliva at pH optimum of 6.8. In contrast to tyrosylprotein sulfotransferase isolated from submandibular salivary glands, salivary enzyme does not require the presence of Triton X-100, NaF and 5'AMP for maximal activity. Similar to the submandibular TPST, the enzyme from saliva also required MnCl(2) for its activity. Maximum TPST activity was observed at 20 mM MnCl(2). The enzyme from saliva was immunoprecipitated and purified by immunoaffinity column using anti-TPST antibody. Affinity purified salivary TPST showed a single band of 50-54 kDa. This study is the first report characterizing a tyrosylprotein sulfotransferase in a secretory fluid. PMID- 16244710 TI - Caleb Hillier Parry (1755-1822): clinician, scientist, friend of Edward Jenner (1749-1823). AB - This article summarizes briefly the life and work of Dr Caleb Hillier Parry (1755 1822), a friend of Dr Edward Jenner. He made original clinical observations, including the bradycardic effect of carotid artery compression, and the association of thyroid enlargement with cardiac disease. He also undertook experimental work and published extensively, including the first book devoted to angina. PMID- 16244711 TI - The influence of Sir Andrew Clark (1826-93) on William Osler (1849-1919). AB - Three lines of evidence suggest that Sir Andrew Clark had a significant influence on William Osler's postgraduate training: (1) numerous references to Clark in Osler's textbook of medicine; (2) the similarity of Clark's and Osler's aphoristic messages for students; and (3) personal encounters, including Clark's reminiscence that he 'had striven ten years for bread, ten years for bread and butter, and twenty years for cakes and ale'. Clark's meticulous approach to clinical problems, his work ethic and his enthusiasm for bedside teaching may have fostered or reinforced these same attributes in Osler. Osler honoured Clark's memory by helping Sir John MacAlister and others realize Clark's ambition to unite various London medical societies into a single organization, which, in 1907, became the Royal Society of Medicine. PMID- 16244712 TI - George E Holtzapple (1862-1946) and oxygen therapy for lobar pneumonia: the first reported case (1887) and a review of the contemporary literature to 1899. AB - A commemorative plaque in York Hospital in Pennsylvania, USA, records that George E Holtzapple MD is the physician 'who discovered the use of oxygen for the treatment of pneumonia on March 6, 1885'. This paper suggests that Dr Holtzapple was not the first to use oxygen for pneumonia patients but was the first to publish a case report with a reasoned physiological explanation of oxygen therapy. His publication was intended, in his own words, to benefit 'average country practitioners' who had no other means of learning about this valuable therapy. PMID- 16244709 TI - Identification of the moving junction complex of Toxoplasma gondii: a collaboration between distinct secretory organelles. AB - Apicomplexan parasites, including Toxoplasma gondii and Plasmodium sp., are obligate intracellular protozoa. They enter into a host cell by attaching to and then creating an invagination in the host cell plasma membrane. Contact between parasite and host plasma membranes occurs in the form of a ring-shaped moving junction that begins at the anterior end of the parasite and then migrates posteriorly. The resulting invagination of host plasma membrane creates a parasitophorous vacuole that completely envelops the now intracellular parasite. At the start of this process, apical membrane antigen 1 (AMA1) is released onto the parasite surface from specialized secretory organelles called micronemes. The T. gondii version of this protein, TgAMA1, has been shown to be essential for invasion but its exact role has not previously been determined. We identify here a trio of proteins that associate with TgAMA1, at least one of which associates with TgAMA1 at the moving junction. Surprisingly, these new proteins derive not from micronemes, but from the anterior secretory organelles known as rhoptries and specifically, for at least two, from the neck portion of these club-shaped structures. Homologues for these AMA1-associated proteins are found throughout the Apicomplexa strongly suggesting that this moving junction apparatus is a conserved feature of this important class of parasites. Differences between the contributing proteins in different species may, in part, be the result of selective pressure from the different niches occupied by these parasites. PMID- 16244713 TI - Bard-Parker scalpels. PMID- 16244714 TI - Edinburgh medical students peculiarly described as 'occasional auditors'. AB - The phrase 'occasional auditor' appears to have been coined by a Royal Commission of 1826-32 to describe students who attended classes but did not intend to graduate. 'Occasional' meant 'infrequent', 'auditor' meant 'listener', and the Commission paid scant attention to these students. At the Edinburgh Medical School, however, such non-graduating students significantly outnumbered the graduates, and this can be shown to have been the case from as early as 1726 and to have continued until 1858, particular attention being paid here to the last two decades (i.e. 1839-58). There is no existing Edinburgh University list of 'occasional auditors', but a methodology of cross-referencing is described which makes it possible to identify the 'occasional auditors' by name. 'Occasional auditors' in the Faculty of Medicine can now be redefined as serious students who had assembled their own course of study in order to further their personal careers, and it is possible to show that very many of these careers were in medicine, largely in the emerging field of general practice. PMID- 16244715 TI - Caleb Hillier Parry (1755-1822). PMID- 16244716 TI - Genealogy of John and Charles Bell: their relationship with the children of Charles Shaw of Ayr. AB - The Reverend William Bell had six children who survived infancy. Two of his sons entered the legal profession and two other sons became distinguished anatomists and surgeons--John Bell, said for 20 years to have been the leading operating surgeon in Britain and throughout the world--and Sir Charles Bell, possibly the most distinguished anatomist and physiologist of his day. Information is not known about the fifth son or their sister. Charles Shaw, a lawyer of Ayr, had four sons and two daughters who survived infancy. Two of his sons, John and Alexander, became anatomists and later surgeons at the Middlesex Hospital, and both worked closely with Charles Bell at the Great Windmill Street School of Anatomy. His third son entered the law and his fourth son became a distinguished soldier. The two daughters of Charles Shaw married into the Bell family: Barbara married George Joseph Bell and Marion married Mr (later Sir) Charles Bell. PMID- 16244717 TI - The death of George Washington (1732-99) and the history of cynanche. AB - George Washington died in the winter of 1799 from acute epiglottitis during an epidemic of influenza. The details of the illness were fully recorded by his secretary, Tobias Lear, and this is the first published description in English of this condition. An account is given of the medical treatment and controversies that arose in criticism of the attendant doctors. PMID- 16244718 TI - Maud Gonne MacBride (1866-1953): an indomitable consumptive. AB - Maud Gonne MacBride was an English debutante who became an Irish rebel and muse to William Butler Yeats. Wealth did not protect her or her family from tuberculosis but, in spite of frequent relapses and three pregnancies, she invariably recovered to lead an active, fruitful life until her death in 1953 at the age of 86. She refused treatment with tuberculin and the induction of an artificial pneumothorax but went through a course of creosote inhalations. Her indomitable spirit helped her to outwit the 'Captain of these Men of Death' at a time when he fully deserved that title. PMID- 16244719 TI - Alfred Salter (1873-1945). PMID- 16244721 TI - Subphthalocyanines as fluoro-chromogenic probes for anions and their application to the highly selective and sensitive cyanide detection. AB - The use of a subphthalocyanine derivative as a selective chromo-fluorogenic reporter for the anion cyanide in mixed aqueous solutions is reported. PMID- 16244722 TI - Contra-Hofmeister anion extraction by cyclosteroidal receptors. AB - Steroid-based receptors with enclosed binding sites, formed from quaternary ammonium and macrocyclic bis-urea units, can substantially override the Hofmeister series in anion phase transfer experiments. PMID- 16244720 TI - Polymeric electrochromics. AB - Conducting polymers are excellent candidates for applications in displays, mirrors, windows, light-emitting diodes, photovoltaics, near-infrared devices and electrochromic devices. From these potential applications, in this article, we will focus on the electrochromic polymers and devices. Although several objective studies have been conducted in the last decade, bringing to light many advantages over other types of electrochromics, polymeric electrochromics have not yet received the industrial attention that they deserve. One of the most important and dazzling advantages of polymers over the other types of electrochromics is the ease of modification of a polymer's backbone, that changes almost the entire properties of the material and switches many disadvantages into advantages. Our recent completion of the deficient third leg of additive primary colour space was a very good example of tailoring the polymer backbone. This discovery could be considered as one of the milestones of commercialization of polymeric electrochromics. In this article, we will also discuss the completion of the additive primary colours, red, green and blue (RGB), in polymeric electrochromics and their ways of commercialization. PMID- 16244723 TI - Two-color luminescence from a tetranuclear Ir(III)/Ru(II) complex. AB - A new tetranuclear compound containing Ru(II) and Ir(III) polypyridine subunits exhibits two independent emissions at room temperature, as a consequence of weak interchromophoric coupling; in contrast, at 77 K energy transfer from Ir-based chromophores to the Ru-based ones is quantitative. PMID- 16244724 TI - Selective steroid recognition by a partially bridged resorcin[4]arene cavitand. AB - The partially bridged resorcin[4]arene cavitand featuring a cleft-shaped recognition site formed by two anti-quinoxaline bridges and four convergent HO groups was prepared in three steps and characterised by X-ray crystallography; cavitand was found to be a selective receptor for steroidal substrates in CDCl3, with the best binding observed for steroids with a flat A-ring and two H-bonding sites on rings A and C/D. PMID- 16244725 TI - Organic crystals absorb hydrogen gas under mild conditions. AB - We have studied the hydrogen sorption on three well-known organic hosts that possess vacant lattice voids large enough to accommodate H2 molecules. PMID- 16244726 TI - A highly active catalyst for CO oxidation at 298 K: mononuclear AuIII complexes anchored to La2O3 nanoparticles. AB - Mononuclear La2O3-supported AuIII complexes synthesised from AuIII(CH3)2(C5H7O2) and characterised by X-ray absorption spectroscopy are highly active, stable CO oxidation catalysts at room temperature, demonstrating the importance of the support in stabilizing catalytically active gold species, which need not include zerovalent gold for high activity. PMID- 16244727 TI - PNA forms an i-motif. AB - A C-rich PNA hexanucleotide, p(C5T), has been shown to form an i-motif by nanoelectrospray ionization mass spectrometry coupled with H/D exchange, to have thermal stability comparable with its DNA analogue, but to exist over a much narrower pH range. PMID- 16244728 TI - Enantioselective conjugate addition of phenylboronic acid to enones catalysed by a chiral tropos/atropos rhodium complex at the coalescence temperature. AB - A highly enantioselective rhodium-catalysed conjugate addition of phenylboronic acid to cyclic enones has been achieved using a dynamic library of chiral phosphorus ligands; the tropos/atropos nature of the ligands in the rhodium complex has been characterised via 31P-NMR. PMID- 16244729 TI - Regiochemical control of the catalytic asymmetric hydroboration of 1,2 diarylalkenes. AB - The hydroboration of stilbenes and related disubstituted alkenes catalysed by QUINAP complexes may proceed with high enantio- and regioselectivity; rhodium and iridium catalysts give the same product regioisomer but opposite enantiomers. PMID- 16244730 TI - Well-controlled reversible addition-fragmentation chain transfer radical polymerisation under ultraviolet radiation at ambient temperature. AB - Controlled reversible addition-fragmentation chain transfer radical polymerisation of methyl acrylate was carried out under long-wave (lambda > or = 365 nm) ultraviolet radiation using an acylphosphine oxide as a photoinitiator at ambient temperature; the polymerisation shows a "living" character at high conversions of polymerisation and leads to well-defined polymers with narrow polydispersities (Mw/Mn < 1.1). PMID- 16244731 TI - Phase identification and quantification in a devitrified glass using homo- and heteronuclear solid-state NMR. AB - A complex mixture resulting from the devitrification of an aluminophosphate glass has been studied for the first time using a combination of homo- and heteronuclear solid-state NMR sequences that offers the advantage of subsequent quantification. PMID- 16244732 TI - Assembled bright green fluorescent zinc coordination polymer. AB - A 1D zinc coordination polymer with bright green fluorescence, [Zn(4,4' bipy)(H2O)4][Zn(4,4'-bipy)(1.5)(L)(H2O)2]2(L).6H2O (L = 4,4'-bis(2 sulfonatostyryl)biphenyl), has been designed, hydrothermally synthesized and characterized; the interesting structure consists of three types of infinite chain: a neutral sandwich-like chain, ionic and cationic chains. PMID- 16244734 TI - Ag/SiO2: a novel catalyst with high activity and selectivity for hydrogenation of chloronitrobenzenes. AB - Ag/SiO2 prepared by an in situ reduction method are found, for the first time, to be highly effective and recyclable catalysts for the selective hydrogenation of a range of chloronitrobenzes to their corresponding chloroanilines, which are of great potential as industrially viable and cheap novel catalysts for the production of chloroanilines. PMID- 16244733 TI - A geometric switching approach toward thermal activation of metalloenediynes. AB - Tetradentate metalloenediynes with strong imine and weaker thioether coordination serve as a geometrically non-rigid switch to drive thermal Bergman cyclization. PMID- 16244735 TI - Ortho-hydroxylation of benzoic acids with hydrogen peroxide at a non-heme iron center. AB - The iron-assisted hydroxylation of benzoic acid to salicylic acid by 1/H2O2 has been achieved in good yield under mild conditions (where is [Fe(II)(BPMEN)(CH3CN)2](ClO4)2 and BPMEN =N,N'-dimethyl-N,N'-bis(2 pyridylmethyl)ethane-1,2-diamine); the product of this reaction is a novel mononuclear iron(III) complex with a chelating salicylate. PMID- 16244736 TI - The direct alpha-zincation of amides, phosphonates and phosphine oxides by H-Zn exchange. AB - Stoichiometric or catalytic quantities of simple 2 degrees amines greatly increase the rate of H-Zn exchange between ZnPh2 and a range of relatively non acidic substrates, allowing for the convenient and direct preparation of alpha functionalized organozincs. PMID- 16244737 TI - Efficient dynamic kinetic resolution of secondary amines with Pd on alkaline earth salts and a lipase. AB - Combination of Pd, supported on alkaline earth type supports with a lipase results in a selective catalytic system for dynamic kinetic resolution of benzylic amines. PMID- 16244738 TI - A catenane consisting of a large ring threaded through both cyclic units of a handcuff-like compound. AB - The template effect of copper(I) allowed the preparation of a new catenane constructed around a bis-macrocyclic unit; a double ring-closing metathesis reaction afforded the central ring which is threaded through both rings of the bis-macrocycle. PMID- 16244739 TI - A fluorescent chemosensor for wide-range pH detection. AB - Simple polyamines, L1-L3, bearing anthracene and benzophenone units at the respective ends, behave as a fluorescent pH sensor applicable to wide-range pH detection. PMID- 16244740 TI - A fluorescent molecular logic gate with multiply-configurable dual outputs. AB - A simple molecule, L, diethylenetriamine bearing anthracene fragments at both ends, behaves as a fluorescent molecular logic gate with "multiply-configurable dual outputs", capable of demonstrating five different logic functions operated by proton (H+) and transition metal cations (Mn+) as inputs. PMID- 16244741 TI - Bis-(hydroxyamino)triazines: highly stable hydroxylamine-based ligands for iron(III) cations. AB - Bis-(hydroxyamino)triazines (BHTs) constitute a new, general and highly versatile group of tridentate iron(III) chelating agents exhibiting higher affinity to iron(III) than other tridentate iron(III) chelators and superior iron(III) over iron(II) selectivity compared to desferrioxamine-B (DFO), EDTA as well as other tridentate ligands. PMID- 16244742 TI - NaNO2-activated, iron-TEMPO catalyst system for aerobic alcohol oxidation under mild conditions. AB - FeCl3-TEMPO-NaNO2 catalyses the selective and mild aerobic oxidation of a broad range of alcohols to the corresponding aldehydes and ketones. PMID- 16244743 TI - Synthesis of protein-silica hybrid hollow particles through the combination of protein catalysts and sonochemical treatment. AB - Hollow spherical particles with protein-silica hybrid shell structures have been synthesized through a combination of the catalytic activity of the protein and sonochemical treatment; the morphologies of the particles were controlled by varying the protein concentration. PMID- 16244744 TI - Chemical synthesis of PEDOT nanofibers. AB - A one-step, room-temperature method is described to chemically synthesize bulk quantities of microns long, 100-180 nm diameter nanofibers of electrically conducting poly(3,4-ethylenedioxythiophene)(PEDOT) in the form of powders, or as optically transparent, substrate-supported films using a V2O5 seeding approach. PMID- 16244745 TI - Anodic aluminium oxide catalytic membranes for asymmetric epoxidation. AB - Catechol-functionalized (salen)Mn complexes can be supported on mesoporous anodized aluminium oxide disks to yield catalytic membranes that are highly active in the enantioselective epoxidation of olefins when being deployed in a forced-through-flow reactor. PMID- 16244746 TI - Combination of ring-opening polymerization and "click" chemistry towards functionalization of aliphatic polyesters. AB - Azide pendent groups of aliphatic polyesters have been derivatized into tertiary amines, ammonium salts and poly(ethylene oxide) grafts. The experimental conditions have been optimized (organic solvent, 35 degrees C), such that the aliphatic polyesters are not degraded, including even poly(lactide) which is very sensitive to attack by weak nucleophiles. PMID- 16244748 TI - Improved method of the MALDI-TOF analysis of DNA with nanodot sample target plate. AB - We report a new sample target plate for MALDI-TOF mass spectrometry made of SiO2 with 30 nm Pt dots for which a highly reproducible and improved DNA analysis was achieved. PMID- 16244747 TI - A novel calamitic mesophase semiconductor with the fastest mobility of charged carriers: 1,4-di(5'-octyl-2'-thienyl)benzene. AB - The carrier mobility of highly ordered lamellar mesophases was evaluated by a Time-of-Flight (TOF) method for mesogenic 1,4-di(5'-octyl-2'-thienyl)benzene (8 TPT-8) to reveal the fastest drift mobility (0.1 cm(2) V(-1) s(-1) for the lowest temperature mesophase) in calamitic liquid crystals reported to date. PMID- 16244749 TI - Alkanes-assisted low temperature formation of highly ordered SBA-15 with large cylindrical mesopores. AB - Highly ordered SBA-15 silicas with large cylindrical mesopores (approximately 15 nm) are successfully obtained with the help of NH4F by controlling the initial reaction temperatures in the presence of excess amounts of alkanes. PMID- 16244750 TI - A versatile organocatalyst for the asymmetric conjugate addition of nitroalkanes to enones. AB - 5-Pyrrolidin-2-yltetrazole performs as an improved catalyst for the asymmetric addition of a range of nitroalkanes to cyclic and acyclic enones, with good to excellent enantioselectivity. PMID- 16244751 TI - Nanotube brushes: polystyrene grafted covalently on CNx nanotubes by nitroxide mediated radical polymerization. AB - Polymer brushes consisting of polystyrene (PS) chains bonded covalently to N doped multiwalled carbon nanotubes (CNx) were synthesized by a "grafting from" route using nitroxide mediated radical polymerization (NMRP). PMID- 16244752 TI - Conformational and spacial preferences for substrates of PepT1. AB - The conformation at the first residue of dipeptide substrates for the peptide transporter PepT1 has been probed using constrained peptide analogues, and the active conformation has been identified. PMID- 16244753 TI - Selective electrochemical sensing of acidic organic molecules via a novel guest to-host proton transfer reaction. AB - Ferrocene-containing amidopyridine receptors bind carboxylic acids and the amino acid phenylalanine in acetonitrile via a novel proton transfer process that enables guests to be electrochemically sensed by positive shifts in the ferrocene centred redox potentials. PMID- 16244754 TI - Changing management of clinical low-stage testicular cancer. AB - Stage I and II testicular germ cell tumors (GCTs) are almost always cured with appropriate treatment and most ongoing research regarding these tumors focuses on minimizing treatment toxicity. The management of clinical stage I testicular GCTs has grown more complicated due to the emergence of a brief course of chemotherapy as an additional treatment option for stage I seminomas and stage I nonseminomas. In addition, growing concern about radiation-induced cancers and other late toxicity has dulled enthusiasm for radiotherapy as a treatment for stage I seminomas. However, recent randomized trials have shown that radiotherapy doses and field sizes can be lowered without compromising cure rates and it is possible that this reduction in radiation exposure will reduce the rate of secondary cancers. At this point in history, stage I patients have three treatment options following radical orchiectomy: adjuvant (sometimes called "primary") chemotherapy (carboplatin for seminomas and the combined regimen of bleomycin, etoposide, and cisplatin for nonseminomas), surveillance, and either retroperitoneal lymph node dissection (for nonseminomas) or radiotherapy (for pure seminomas). Clinical studies have made it possible to identify subgroups of patients at high and low risk for relapse and this has made it possible to tailor treatment decisions to the individual patient's postorchiectomy relapse risk. PMID- 16244755 TI - Evaluation of stature development during childhood and adolescence in individuals with familial hypophosphatemic rickets. AB - This review was conducted to study the diagnosis, treatment, and growth progression in infants and adolescents with familial hypophosphatemic rickets. The bibliographic search was carried out utilizing the electronic databases MEDLINE, OVID, and LILACS and by direct research within the last 15 years using the keywords rickets, familial hypophosphatemia, vitamin D deficiency, stature growth, childhood, and adolescence. Article selection was done by comparing the evaluation of the growth in patients with familial hypophosphatemic rickets, including the variables that might affect them, for possible future therapeutic proposals. It is concluded that the most significant fact in the treatment of familial hypophosphatemic rickets in infancy was the magnitude of the final stature. The use of growth hormone can be helpful in these patients. However, research reporting treatments with the use of the growth hormone for rickets are controversial. The majority of the authors agree that treatment using vitamin D and phosphate enables some statural growth in cases of early diagnosis, reflecting a better prognosis. PMID- 16244757 TI - Health-related quality of life and hearing aids: a tutorial. AB - Health-related quality-of-life (HRQoL) instruments measure the impact of a disorder and treatment on several attributes that are thought to constitute the self-perceived health status of an individual. This tutorial reviews the conceptual framework of HRQoL, including the challenges associated with defining and measuring HRQoL, specifically as it applies to audiologic care. A relatively new instrument, the World Health Organization-Disability Assessment Schedule II, will be discussed as a potentially valuable instrument to measure the impact of hearing loss and hearing aid intervention on self-perceived HRQoL. PMID- 16244760 TI - Generic low molecular weight heparins: a significant dilemma. PMID- 16244758 TI - The WHO-DAS II: psychometric properties in the measurement of functional health status in adults with acquired hearing loss. AB - The World Health Organization's (WHO) Disability Assessment Scale II (WHO-DAS II) is a generic health-status instrument firmly grounded in the WHO's International Classification of Functioning, Disability and Health (WHO-ICF). As such, it assesses functioning for six domains: communication, mobility, self-care, interpersonal, life activities, and participation. Domain scores aggregate to a total score. Because the WHO-DAS II contains questions relevant to hearing and communication, it has good face validity for use as an outcome measure for audiologic intervention. The purpose of the present study was to determine the psychometric properties of the WHO-DAS II on a sample of individuals with adult onset hearing loss, including convergent validity, internal consistency, and test retest stability. Convergent validity was established by examining correlations between the WHO-DAS II (domain and total scores) and the Abbreviated Profile of Hearing Aid Benefit (APHAB) and the Hearing Aid Handicap for the Elderly (HHIE), two disease-specific measures, as well as with the Short Form-36 for veterans (SF 36V), a second generic measure. Data on all four measures were collected from 380 older individuals with adult-onset hearing loss who were not hearing aid users. The results of the convergent validity analysis revealed that the WHODAS II communication domain score was moderately and significantly correlated with scores on the APHAB and the HHIE. WHO-DAS II interpersonal and participation domain scores and the total scores were also moderately and significantly correlated with HHIE scores. These findings support the validity of using the WHO DAS II for assessing activity limitations and participation restrictions of adult onset hearing loss. Several WHO-DAS II domain scores and the total score were also significantly and moderately-markedly correlated with scores from the SF 36V. These findings support the validity of the WHO-DAS II as a generic health status instrument. Internal consistency reliability for all the domain scores was adequate for all but the interpersonal domain. Test-retest stability for all the domain scores was adequate. Critical difference values were calculated for use in clinical application of the WHO-DAS II. From these findings, we concluded that the WHO-DAS II communication, participation, and total scores can be used to examine the effects of adult-onset hearing loss on functional health status. Further work examining the utility of the WHO-DAS II as an outcome measure for hearing aid intervention is warranted. PMID- 16244761 TI - Argatroban and lepirudin: clinical trials to clinical practice. PMID- 16244759 TI - The WHO-DAS II: measuring outcomes of hearing aid intervention for adults. AB - The World Health Organization's Disability Assessment Scale II (WHO-DAS II) is a generic health-status instrument that provides six domain scores and a total, aggregate score. Two of the domain scores, communication and participation, and the total score, have good validity, internal-consistency reliability, and test retest stability in individuals with adult-onset hearing loss. As such, these two domain scores and the total WHO-DAS II score may be useful as generic outcome measures to assess the effectiveness of hearing aid intervention for this population. Before the use of the WHO-DAS II in hearing aid clinical trials, however, the responsiveness of the instrument and the short- and long-term outcomes to hearing aid intervention had to be determined. Responsiveness and outcomes were assessed in 380 veterans (approximately half received hearing aids and half served as controls) by examining group differences, effect-size estimates, and individual differences as a function of hearing aid intervention. For comparison, data also were obtained on two disease-specific measures, the APHAB and the HHIE. The WHO-DAS II communication domain and total scores were sufficiently responsive to hearing aid intervention for use in future studies in which group differences are to be detected. The WHO-DAS II participation domain was not sufficiently responsive to hearing aid intervention. The APHAB and HHIE, both disease-specific measures, were more sensitive to hearing aid intervention than the generic measure. The short- and long-term outcomes of hearing aid intervention were also examined in the present study. Group outcomes for hearing aid intervention can be expected to be stable for at least 6 months when measured by WHO-DAS II total score and for at least 12 months when measured by the WHO-DAS II communication domain scores. Effect-size estimates and examination of the number of individuals exhibiting change scores exceeding 90% critical differences for true changes in scores indicate that for clinical applications, disease specific instruments are more useful than the WHO-DAS II. The findings of this study support the use of the WHO-DAS II as a generic measure in hearing aid trials research so as to allow for comparisons of health-status outcomes across different diseases or disorders. PMID- 16244762 TI - A comparison of lepirudin and argatroban outcomes. AB - Although both argatroban and lepirudin are used for the management of heparin induced thrombocytopenia (HIT), data comparing these agents are lacking. The objective of this project was to compare the clinical outcomes of lepirudin vs argatroban therapy. Patients who received a direct thrombin inhibitor (DTI) from January 2000 to December 2001 were identified. Medical charts were retrospectively reviewed and relevant data extracted. The primary efficacy outcome was effective anticoagulation and the primary safety outcome was major bleeding. Data were analyzed using the t test and Fisher's exact test. Sixty-one lepirudin patients and 29 argatroban patients received a DTI during the study period. A new diagnosis of HIT was the indication for DTI therapy in 44.8% of argatroban patients and 57.4% of lepirudin patients. Effective anticoagulation was achieved in 77.8% of argatroban patients and 69.5% of lepirudin patients (p = .61). Major bleeding occurred in 10.3% and 11.5% of argatroban and lepirudin patients, respectively (p = 1.0). Argatroban and lepirudin demonstrated comparable safety and efficacy outcomes. PMID- 16244763 TI - Associations of thrombophilia, hypofibrinolysis, and retinal vein occlusion. AB - We prospectively assessed whether thrombophilia and hypofibrinolysis, amplified by thrombophilic hormone replacement therapy (HRT), were associated with retinal vein occlusion (RVO). We studied 44 cases (18 men, 26 women), > or = 3 months after RVO, 42 with central RVO, 2 with branch RVO, in the consecutive order of their referral by 2 community-based ophthalmologists. PCR and serologic coagulation assays were compared to 83 and 40 healthy adult normal controls, respectively. The 4G allele frequency of the plasminogen activator inhibitor-1 (PAI-1) gene, associated with hypofibrinolysis, was 56 of 88 (64%) in cases vs 79 of 166 (48%) in controls, X(2) = 5.95, p = .015. The PAI-1 gene product, plasminogen activator inhibitor activity (PAI-Fx), was higher in cases than controls (age-race-sex- adjusted mean 12.2 U/mL vs 6.3, p = .013). By stepwise logistic regression, the PAI-1 gene 4G allele was associated with RVO, odds ratio 1.94, 95% CI 1.12-3.34, p = .018. Thrombophilic resistance to activated protein C (RAPC) was present in 6 of 32 (19%) of cases vs 0 of 40 (0%) controls, Fisher's p [p(f)] = .006. Thrombophilic high factor VIII (> 150%) was present in 3 of 30 (10%) cases vs 0 of 40 (0%) controls, p = .041, p(f) = .07. Comparing 23 RVO cases < or = age 55 and controls < or = age 55 (n = 44 for PCR, n = 40 for serologic measures), RAPC was present in 17% of cases vs 0% controls (p(f) = .026), high Factor VIII in 17% vs 0% (p(f) = .026), heterozygosity for the G1691A Factor V Leiden mutation in 13% vs 2% (p(f) = 0.11), and the 4G allele frequency of the PAI-1 gene 74% vs 39% (p = .0001). PAI-Fx was higher in cases than controls (age-race-sex adjusted mean 12.7 U/mL vs 6.7, p = .016). The case control odds ratio for the PAI-1 4G allele was 5.54, 95% CI = 1.86-16.7, p = .002. Of the 26 women, 9 (35%) took HRT; 4 of the 9 had PAI-1 gene 4G4G homozygosity, 2 had thrombophilic high anticardiolipin antibody (IgG), 1 was heterozygous for the G1691A Factor V Leiden mutation, and 2 were heterozygous for the thrombophilic PL A1/A2 mutation of the platelet glycoprotein IIb/IIIa gene. Associations between heritable coagulation disorders and RVO, most marked in cases < or = age 55, and often amplified in women by thrombophilic HRT, are, speculatively, causal. PMID- 16244764 TI - Elevated plasma levels of fibrin degradation products by granulocyte-derived elastase in patients with disseminated intravascular coagulation. AB - Plasma levels of granulocyte-derived elastase (GE-XDP), D-dimer, and soluble fibrin (SF) were examined in 177 patients with disseminated intravascular coagulation (DIC) of various etiologies. Plasma levels of GE-XDP and D-dimer, but not SF, were significantly high in patients with sepsis and solid cancer. The ratio of GE-XDP/ D-dimer was significantly high in patients with trauma, burn, and sepsis, suggesting that fibrinolysis due to GE-XDP may be dominant in DIC. Plasma levels of GE-XDP and D-dimer, but not SF, were significantly high in patients with overt DIC and correlated with DIC score. Plasma levels of GE-XDP, but not SF, correlated significantly with D-dimer. Plasma levels of D-dimer, but not SF, correlated significantly with plasmin plasmin inhibitor complex (PPIC). Plasma levels of GE-XDP and D-dimer, but not SF, were significantly high in nonsurvivors. Plasma levels of GE-XDP, but not SF, correlated significantly with sepsis-related organ failure assessment (SOFA) score. These results suggest that GE-XDP is a potentially useful marker for the diagnosis of overt-DIC and as a predictor of organ failure-related outcome. PMID- 16244765 TI - Control of bleeding caused by thrombocytopenia associated with hematologic malignancy: an audit of the clinical use of recombinant activated factor VII. AB - This paper presents an analysis of 24 cases in which recombinant factor VIIa (rFVIIa) was used in the management of hemorrhage in patients with thrombocytopenia associated with hematologic malignancies. This is the largest case aggregation to date and focuses on preliminary experience in the off-label use of this hemostatic agent. Data were extracted from the international, Internet-based registry, www.haemostasis.com, accessed in September 2003. The search results were manually cross-checked against monthly summary reports. The physicians providing the cases were contacted individually to approve the use of their cases, supply any information missing from the database, and validate the data already held. Patients with acute myeloid leukemia, acute lymphoblastic leukemia, Hodgkin's disease, non-Hodgkin's lymphoma, Burkitt's lymphoma, B-cell or T-cell lymphoma, or aplastic anemia received rFVIIa at total doses of between 18 and 1040 mug/kg body weight. Bleeding stopped in 11 of 24 (46%) patients, markedly decreased in 8 of 24 (33%) patients, and decreased in 4 of 24 (17%) patients. In most patients, the response was achieved within 2.5 hours of administration of rFVIIa. The use of rFVIIa was generally well tolerated -- 1 case of ischemic stroke was considered to be possibly related to rFVIIa administration, but this has yet to be confirmed. A review of these 24 cases submitted to the www.haemostasis.com database suggests that rFVIIa is beneficial in the management of hemorrhage in patients with thrombocytopenia and hematologic malignancies. This warrants further investigation in rigorously controlled clinical trials. PMID- 16244766 TI - Increased tyrosine phosphorylation of platelet proteins including pp125(FAK) suggests endogenous activation and aggregation in pulmonary hypertension. AB - Despite of several lines of evidence indicating a pathophysiologic role of platelets in pulmonary hypertension, the occurrence of chronic endogenous platelet activation has been a matter of debate. It was hypothesized that the pattern of tyrosine phosphorylation of platelet proteins examined ex vivo could provide information on the state of platelet activation. This was examined in 10 patients with pulmonary arterial hypertension aged 18 to 53 years. Phosphotyrosine density and the amounts of specific proteins were analyzed in resting platelets after reaction with anti-phosphotyrosine, anti-pp60(s-src), anti-pp125(FAK), and anti-alphaIIbbeta3 antibodies. There was a 79% increase in protein-associated phosphotyrosine in patients in comparison to levels in controls (p<0.05). In particular, phosphorylation on tyrosine residues of pp120 and pp125(FAK) increased 24% and 57%, respectively (p<0.05). Although pp60(s-src) associated phosphotyrosine was not altered in the patient group as a whole, it was clearly decreased in three subjects. Platelet content of beta3 integrin, pp60(s-src), and pp125(FAK), was not altered. This pattern of phosphorylation suggests an ongoing process of platelet activation. Because phosphorylation of pp125(FAK) is a late, integrin-dependent event, results suggest that platelet activation and aggregation occur in vivo in these patients. PMID- 16244768 TI - Gefitinib affects functions of platelets and blood vessels via changes in prostanoids balance. AB - Prostaglandins (PGs) and thromboxane (TX) produced by cyclooxygenase (COX) have a great influence on vascular systems and platelet functions. The serum levels of epidermal growth factor (EGF) and PGs were measured in patients with lung cancer treated with gefitinib, and the influence of EGF on platelet aggregation was investigated. Twenty patients were investigated. The serum level of TXB(2) increased significantly in all patients who received gefitinib for 2 weeks (before vs. after = 94.1 +/- 47.3 vs. 190.9 +/- 54.3, p<0.01). TXB(2) also increased significantly in responders without concurrent chemotherapy (before vs. after = 79.3 +/- 35.5 vs. 194.5 +/- 58.1, p<0.05), but not in non-responders (before vs. after = 106. 5 +/- 65.8 vs. 162.2 +/- 52.8, N.S.). PG 6-keto F1alpha and PGE(2) did not exhibit significant changes. Furthermore, EGF showed no significant change (after vs. before = 234 +/- 35 vs. 276 +/- 72, N.S.). Although there was no correlation between the levels of EGF and TXB(2) (N.S.), the PG 6 keto F2alpha/TXB(2) ratio decreased significantly (before vs. after = 0.054 +/- 0.018 vs 0.033 +/- 0.015, p<0.05). The secondary platelet aggregation observed after high-dose adenosine diphosphate stimulation was inhibited after a 1-minute preincubation with EGF. Platelet aggregation in patients after gefitinib administration tended to accelerate and secondary aggregation was observed after low-dose adenosine diphosphate stimulation. We conclude that careful observation is needed for patients with chronic obstructive pulmonary disease, pulmonary fibrosis, and thromboembolic diseases receiving gefitinib. Furthermore, measurement of prostanoids may be a good predictor of the beneficial and adverse effects. Moreover, the combination of gefitinib with a COX inhibitor that regulates TXA(2)/PGI(2) balance should be evaluated. PMID- 16244767 TI - Different effect of statins on platelet oxidized-LDL receptor (CD36 and LOX-1) expression in hypercholesterolemic subjects. AB - Hydroxymethyl-glutaryl-CoA-reductase inhibitors (statins) reduce cardiovascular mortality by decreasing cholesterol as well as by non-lipid-related actions. Oxidized low-density lipoproteins (ox-LDL) are pro-atherogenic molecules and potent platelet agonists. CD36 and lectin-like ox-LDL receptor-1 (LOX-1) are specific ox-LDL receptors also expressed in platelets. This study was planned to address whether treatment with atorvastatin 10 mg/day, pravastatin 40 mg/day or simvastatin 20 mg/day could affect platelet CD36 and LOX-1 expression. Twenty four patients for each treatment were evaluated after 3, 6, and 9 days and at 6 weeks for complete lipid profile (chromogenic), ox-LDL (ELISA), platelet P selectin (P-sel), CD36, LOX-1 (FACS), and intracellular citrullin recovery (iCit) (HPLC). Data show hyperactivated platelets (P-sel absolute values, percent variation in activated cells, all p < 0.001), and CD36 and LOX-1 overexpression (all p < 0.001) in patients at baseline. P-sel, CD36, and LOX-1 were significantly decreased by atorvastatin and simvastatin (all p < 0.01) and related with iCit increase (r = 0.58, p < 0.001) and platelet-associated ox-LDL (r = 0.51, p < 0.01) at 9 days. Pravastatin reduced LOX-1 and P-sel (p < 0.05) at 6 weeks in relation with decreased LDL and ox-LDL (r = 0.39, p < 0.01 and r = 0.37, p < 0.01, respectively). These data suggest that atorvastatin and simvastatin reduce platelet activity by exposure of CD36 and LOX-1 before significant LDL reduction, whereas pravastatin action is detected later and in relation with LDL and ox-LDL lowering. Rapid and consistent reduction of CD36 and LOX-1 could be considered a direct anti-atherothrombotic mechanism related to the role of ox-LDL in platelet activation, platelet-endothelium interactions, and NO synthase activity. PMID- 16244769 TI - Endothelial cell activity in chronic obstructive pulmonary disease without severe pulmonary hypertension. AB - Pulmonary hypertension is common in patients with chronic obstructive pulmonary disease (COPD), but the precise mechanism of vascular impairment in these patients is unknown. We, therefore, decided to investigate whether endothelial cell dysfunction is present in patients with COPD with a wide range of chronic airflow obstruction before the development of severe pulmonary hypertension. Selected plasma markers of endothelial cell activity were studied: nitrate+nitrite (NO-(2)/NO-(3)), thrombomodulin (TM), tissue factor pathway inhibitor (TFPI), soluble selectins (endothelium sES, leukocyte sLS, platelet sPS), soluble intercellular adhesion molecule-1 (sICAM-1), and soluble platelet endothelial cell adhesion molecule-1 (sPECAM-1). Twenty-five patients with COPD (forced expiratory volume in one second/vital capacity [FEV(1)/VC] < 88% predicted) and 29 healthy control subjects were recruited to the study. Among patients nine had a pulmonary artery systolic pressure (PASP) between 15 and 30 mmHg, 13 between 32 and 38 mmHg, 2 had a PASP of 41 and 42 mmHg, respectively. One patient had severe pulmonary hypertension with a PASP of 70 mmHg. The average FEV(1) of patients with COPD was 46 +/- 4% predicted. As compared to control subjects, patients with COPD showed a significant increase in plasma levels of TM and TFPI, indicating that their endothelial cells are still able to produce potent coagulation inhibitors. Levels of NO-(2)/NO-(3) were similar in the two groups of subjects examined, further suggesting preserved endothelial function in patients with COPD. In regard to adhesion molecules, patients with COPD showed a reduction in sLS, sPS, and sPECAM-1, and an increase in sICAM-1. This study shows that endothelial cell activity is largely preserved in patients with COPD without severe pulmonary hypertension, suggesting that these patients, despite quite severe airway obstruction, retain reasonably normal endothelial function until they develop severe pulmonary hypertension. PMID- 16244770 TI - Idiopathic intracranial hypertension: associations with thrombophilia and hypofibrinolysis in men. AB - The existence of an association between idiopathic intracranial hypertension (IIH) and coagulation disorders in men was assessed prospectively. Microthrombi, associated with thrombophilia-hypofibrinolysis, occlude arachnoid sinus villi, thus reducing resorption of cerebrospinal fluid, leading to IIH. Ten consecutively referred men with IIH, nine whites, one African American, median age 36 years, were 2 to 1 matched by age and race by healthy male controls. Polymerase chain reaction assays were done for four thrombophilic and one hypofibrinolytic gene mutations: G1691A factor V Leiden, G20210A prothrombin, C677T MTHFR, platelet glycoprotein IIb/IIIa (PL A1/A2), and 4G/5G polymorphism of the plasminogen activator inhibitor (PAI-1) gene promoter. Coagulation measures in plasma included dilute Russel's viper venom time (dRVVT), activated partial thromboplastin time (aPTT), the lupus anticoagulant, factor VIII, factor XI, plasminogen activator inhibitor activity (PAI-Fx), protein C antigenic, protein S total (antigenic), protein S free (antigenic), antithrombin III (functional), and resistance to activated protein C (RAPC). Tests performed on serum included anticardiolipin antibodies, homocysteine, and Lp(a). The body mass index was 40 kg/m(2) or greater (extremely obese) in two men, 30 to 40 kg/m(2) (obese) in three, and was 25 to 30 kg/m(2) in five (overweight). Cases differed from controls for inherited 4G4G homozygosity of the PAI-1 gene, four of 10 (40%) vs. one of 20 (5%), Fisher's p [p(f)]= .031, and for high levels (>21.1 U/mL) of the hypofibrinolytic PAI-1 gene product, PAI-Fx, 5 of 10 (50%) vs. one of 18 (6%), p(f) = .013. Thrombophilic factor VIII was high (> or = 150%) in three of 10 (30%) cases vs. zero of 16 (0%) controls, p(f)=. 046. The thrombophilic lupus anticoagulant was present in two of 10 (20%) cases vs. zero of 32 (0%) controls, p(f) = .052. Heritable hypofibrinolysis and heritable and acquired thrombophilia appear, speculatively, to be treatable etiologies of IIH in men. Understanding contributions of hypofibrinolysis and thrombophilia to the development of IIH should facilitate development of novel new approaches to treat this often disabling neurologic disorder. PMID- 16244771 TI - Plasma levels of thrombin-activatable fibrinolysis inhibitor in primary and secondary thrombocytosis. AB - An elevated platelet count is a common finding in both hospitalized and ambulatory patients. Thrombosis and bleeding complications are more frequently observed in patients with clonal thrombocytosis than secondary thrombocytosis. The aim of this study was to investigate the behaviors of thrombin-activatable fibrinolysis inhibitor (TAFI) activity, the inhibitor of fibrinolysis, and also prothrombin time (PT), active partial thromboplastin time, and D-dimer and fibrinogen levels in 21 patients affected with clonal thrombocytemia as compared with 21 patients with reactive thrombocytosis and 21 healthy controls. In the clonal thrombocytemia group, plasma levels of TAFI activity were significantly higher than in both the reactive thrombocytosis and the control group. Plasma levels of leukocyte and platelet counts were significantly higher in the clonal thrombocytemia group than in the other two groups and also higher in the reactive thrombocytosis group than in the control group, which was also significant. Fibrinogen and D-dimer levels were higher in patients than in the control group but showed no significant difference between the clonal and secondary thrombocytosis groups. Plasma levels of PT and aPTT were higher in secondary thrombocytosis group than the clonal thrombocytosis group. The results of this study showed for the first time that TAFI activity is increased in patients with clonal thrombocytosis. These increased levels in clonal thrombocytosis can be considered a factor to explain the thrombotic tendency in myeloproliferative disorders. PMID- 16244772 TI - Significance of alpha(IIb)beta(3) in subacute stent thrombosis after percutaneous coronary intervention. AB - SUMMARY: Soluble P-selectin and whole blood aggregation (WBA) were measured after percutaneous coronary intervention in patients who then received antiplatelet therapy. One had subacute thrombosis on day 7. This patient's WBA exhibited time dependent enhancement. In addition, the accentuation of WBA on day 3 was observed when anti-alpha(IIb)beta(3) antibody was added. The remaining 22 patients were divided according to WBA results on day 3 with anti-alpha(IIb)beta(3) antibody added (A group, WBA enhanced; B group, WBA did not enhance). WBA on day 3 was similar in the two groups. The ratio of WBA with and without anti alpha(IIb)beta(3) antibody was higher in group A than in group B. A significant time-dependent increase of soluble P-selectin was observed in the A group. These results suggest that enhancement of WBA with anti-alpha(IIb)beta(3) antibody after percutaneous coronary intervention predicts subacute thrombosis. PMID- 16244773 TI - Effects of percutaneous coronary thrombectomy with the X-sizer catheter on epicardial flow and microvascular function in acute coronary syndromes. AB - During percutaneous coronary intervention, slow coronary flow and distal embolization are still important problems, especially in cases with intracoronary thrombus. The aim of this study was to learn the effectiveness and early term results of thrombectomy with the X-SIZER catheter system in acute coronary syndrome. Twenty-nine patients (22 [76%] men; 55.9 +/- 11.1 years) with acute coronary syndrome and intracoronary thrombus detected in coronary angiography were included into the study. X-sizer thrombectomy was applied to 14 of the patients, and conventional percutaneous transluminal coronary angioplasty (PTCA) was applied to the others. Baseline characteristics were similar in both groups. Mean thrombolysis in myocardial infarction (TIMI) flow increased from 0.8 +/- 0.9 to 2.4 +/- 0.6 in X-sizer-treated patients (p<0.001) and TIMI 3 flow was maintained in 71.4% of the patients. Similary, mean TIMI flow increased from 0.36 +/- 0.81 to 2.73 +/- 0.47 in conventional PTCA-treated patients (p<0.001) and TIMI 3 flow was maintained in 73% of the patients (NS). Mean myocardial blush grade (MBG) increased from 0.7 +/- 0.7 to 2.6 +/- 0.6 in X-sizer-treated patients (p<0.001) and from 0.27 +/- 0.65 to 2.36 +/- 0.67 in the conventional PTCA treated patients (p<0.001). Postprocedural MBG 3 was obtained in 64.3% of X-Sizer treated patients and in 45% of controls. Although microvascular function in the thrombectomy-applied patients was found better, there was no significant difference between the two groups. Furthermore it was detected that the use of tirofiban yielded no additional improvement in epicardial and microvascular flow. In acute coronary syndromes, use of X-sizer in addition to primary percutaneous coronary interventions is a safe and relatively effective method in the prevention of distal embolization. PMID- 16244774 TI - Coagulation and fibrinolysis in preeclampsia and neonates. AB - Coagulation and fibrinolysis were determined in 67 Indonesian women admitted to the University Hospitals for delivery in Medan. They were diagnosed to be at term gestation (mean 39.3 +/- 1.1 weeks) with moderate and severe preeclampsia (n=32) and in labor, and 8 had preterm labor (gestation mean 33.5 +/- 2.6 weeks). Twenty seven normal pregnant women in labor (gestation mean 39.7 +/- 1.0 weeks) served as controls. Cord blood from 23 neonates from normal pregnancy and 31 neonates from preeclampsia was also evaluated. Preeclamptic women in labor showed further enhanced coagulation activation (F(1+2)) with raised urokinase-like plasminogen activator (u-PA) activity and reduced plasminogen activator inhibitor-2 (PAI-2) levels. In preterm preeclampsia, significantly reduced antithrombin III (ATIII) and PAI-2 levels with further elevated tissue-type PA (t-PA) antigen and plasminogen activator inhibitor-1 (PAI-1) antigen were seen compared to normal pregnancy. These would suggest a state of enhanced thrombin generation with elevated fibrinolytic/inhibitor proteins in preterm preeclampsia. The reduced PAI 2 levels seen in preeclampsia have been suggested to be associated with reduced placental function. Neonates born to mothers of either normal pregnancy or preeclampsia at term showed similar hemostatic changes with reduced fibrinogen, ATIII, t-PA, u-PA antigen, PAI-1 levels, and coagulation activation compared to their respective maternal plasma levels. No significant differences in hemostatic parameters studied between the neonates of both cohorts were seen, and this would suggest that the neonates were protected from the adverse effects of preeclampsia and their hemostatic system was physiologically balanced. PMID- 16244775 TI - Clinico-hematologic profile of factor XIII-deficient patients. AB - A retrospective analysis of clinico-hematologic parameters of 18 factor XIII deficient patients was carried out. The hematologic tests included activated partial thromboplastin time (APTT), prothrombin time (PT), and clot solubility. Laboratory diagnosis of FXIII deficiency was made where bleeding time, PT, APTT, and thrombin time were normal and the clot solubility test result with 5M urea was positive. Factor XIII level with family screening was performed using commercially available kits. History of prolonged bleeding from the umbilical stump was present in four (22.2%) patients. The most common site of bleeding was the skin (11 of 18 patients). Three patients were given prophylaxis (FFP in two, factor XIII in one). A high prevalence of recurrent abortion in female patients with FXIII deficiency (two of the three patients in this study) was observed. PMID- 16244776 TI - eNOS gene affects red cell deformability: role of T-786C, G894T, and 4a/4b polymorphisms. AB - Plasma viscosity and erythrocyte deformability play a key role in maintaining and regulating microcirculation. In vitro and in vivo studies suggested a role for nitric oxide (NO) in modulating flow-mediated vasodilatation and red blood cell deformability. Impaired NO availability due to mutations in eNOS gene might contribute to the altered haemorheologic state. The aim of this study was to investigate the role of eNOS T-786C, G894T, and 4a/4b polymorphisms in modulating the haemorheologic state in a clinical condition characterized by a microcirculatory disorder. Eighty patients with idiopathic sudden sensorineural hearing loss (ISSHL) and 80 healthy subjects were studied. By using a dominant model of inheritance, we found a significant association between eNOS 894T rare variant and ISSHL (odds ratio [OR] 894TT+GT = 2.08, p = 0.03) after adjustment with traditional vascular risk factors. A higher percentage of altered red cell deformability both in patients and in controls carrying the eNOS rare variants was found in comparison to subjects carrying the wild type. Apart from the disease, eNOS T-786C and G894T polymorphisms independently affected the deformability index (OR, -786CC+TC = 2.81, p = 0.01 and OR, 894TT+GT = 2.5, p = 0.02, respectively), in particular in subjects in whom the contemporary presence of the two rare alleles was observed (OR, -786CC+TC and 894TT+GT combined genotype = 6.9, p<0.0001). Our study documented that eNOS gene affects the red blood cell deformability, so possibly contributing to ISSHL, which may represent a suitable model of microcirculatory disorder. PMID- 16244777 TI - Effects of high-dose methylprednisolone therapy on coagulation factors in patients with acute immune thrombocytopenic purpura. AB - Autoimmune thrombocytopenic purpura (ITP) is a disease that presents with skin and mucous membrane bleeding due to thrombocytopenia. In the literature, there are a few studies about the effect of high-dose steroid therapy on coagulation tests in different diseases, but their results are still controversial. In this study, coagulation parameters were investigated that might have a role in hemostatic compensation in childhood acute ITP before and after high-dose methylprednisolone (HDMP) treatment. The study includes 21 children age 1.5 to 14 years with acute ITP and 21 healthy age-matched control subjects. All patients with acute ITP received HDMP for 7 days. Before and after HDMP treatment (0 and 8 days) prothrombin time, partial thromboplastin time, fibrinogen, Protein C, Protein S, antithrombin III, and the levels of factor II (FII), FV, FVII, FVIII, FIX, FX, FXI, and FXII were studied in all subjects. The results were compared with those of the control group. Pre-treatment Protein C and Protein S levels in the patient group were significantly lower than those in the control groups (p<0.05). Protein S and Protein C levels were significantly improved after HDMP treatment in patient group. There were lower FV, FVII, FX values in the patient group compared to the control groups on admission. There was no difference in AT III and fibrinogen levels before and after treatment. As a result, some changes in the coagulation system associated with thrombocytopenia were observed in patients with acute ITP. These changes may be accepted as compensatory mechanisms to maintain hemostasis. PMID- 16244778 TI - Confirmation of the value of a modified long-distance polymerase chain reaction in the detection of inversion intron 22 in severe hemophilia a: a technical note. AB - Hemophilia A is one of the most common bleeding disorders in man. Approximately half the families with a severe disease have an inversion of the factor VIII gene. The inversion may be detected with a long polymerase chain reaction. This is a simple and reproducible method that may yield satisfactory results in approximately 24 hours. The long-distance polymerase chain reaction amplify three very large amplicons with a very GC-rich region of several kilobases, and is detected with a Expand Long Template DNA polymerase (Roche, Mannheim, Germany). Recently this polymerase has been changed with a new chemical composition. The object of the present method is to standardize the technique using the new Expand Long polymerase. For the new protocol, the cycling conditions, the concentration of nucleotide primers, and the buffer are changed. The need for a rapid response is determined in the case of hemophilia A patients, not only by the desire to reach a proper classification, but also by the urgency to inform the carrier status of the mother or of a female relative. PMID- 16244779 TI - Oral contraceptives, thrombosis and socio-ethical values: an inevitable compromise is needed based mainly on a careful evaluation of potential users. PMID- 16244780 TI - The intra-lingual course of the nerves of the tongue. AB - The intra-lingual course of the nerves of tongue has never been studied in the human by the Sihler method. This technique was applied to six human tongues and allowed coloration of the nerves and illustration of the tongue. The course of the hypoglossal, glossopharyngeal, and lingual nerves was defined. Constant anastomoses between the lingual and hypoglossal nerves, between the glossopharyngeal nerve and the hypoglossal and lingual nerves, were demonstrated and may help explain the "neck-tongue" syndrome. This cartography of the nerves of the tongue allowed the definition of the motor and sensory consequences of tongue surgery. PMID- 16244781 TI - The duodenal fossae: anatomic study and clinical correlations. AB - The aim of this study was to present anatomic macroscopic aspects and the relationship between the duodenum and the posterior abdominal wall. The authors describe anatomic types of peritoneal duodenal fossae and stress some points of surgical importance. Twenty-four cadavers, fixed in formalin, were dissected. Ten peritoneal fossae were given prominence and the authors show the anatomical structures topographically, from a superficial plane to a deep viscerae level. There is usually a complete fusion of the duodenal loop with the posterior parietal peritoneum except the duodenojejunal flexure. The study reveals three right retroduodenal fossae, three left retroduodenal fossae, two inferior duodenal fossae, one left paraduodenal fossa and one superior duodenal fossa. These peritoneal recesses mostly result from an incomplete adhesion of the Treitz's fascia. This work provides some explanation of paraduodenal hernias that represent a rare case of intestinal obstruction. Two cases of these internal hernias are illustrated and their pathophysiology and embryologic basis are discussed. PMID- 16244782 TI - Homocysteine levels are associated with MTHFR A1298C polymorphism in Indian population. AB - An elevated level of homocysteine is an independent risk factor for cardiovascular diseases and is associated with other complex disorders. Homocysteine levels can be elevated due to dietary and/or genetic factors. A majority of Indian population have a low level of vitamin B12 (presumably due to vegetarian diet)--a critical nutritional factor, deficiency of which results in hyperhomocysteinemia. Hence, polymorphisms in the genes responsible for homocysteine metabolism can be perceived to have a greater impact in relation to hyperhomocysteinemia in Indian population. For this reason, the effects of diet and/or methylenetetrahydrofolate reductase (MTHFR) polymorphism were assessed in 200 individuals having varying homocysteine levels. Homocysteine levels were significantly elevated in individuals adhering to a vegetarian diet (P = 0.019) or having MTHFR A1298C polymorphism (P = 0.006). The minor allele frequency (MAF) of MTHFR C677T and A1298C was 0.15 and 0.44 respectively in this cohort. Since the MAF of these polymorphisms differed considerably from Caucasian and other Asian populations, frequencies of these polymorphisms were also determined in more than 400 individuals from different ethnic populations, selected from the entire country based on their geographical location and linguistic lineage, and was found to be similar to that of our cohort. The fact that MTHFR A1298C polymorphism is significantly associated with homocysteine levels, and that the CC genotype is present at a higher frequency in the Indian population, makes it extremely relevant in terms of its potential impact on hyperhomocysteinemia. PMID- 16244784 TI - Accuracy and reliability of sentinel node biopsy in patients with breast cancer. Single centre study with long term follow-up. AB - The aim of our study is to evaluate the frequency of false-negative (FN) sentinel node procedures in patients with breast cancer. A total of 791 breast cancer patients underwent sentinel lymph node (SLN) biopsy at our institution between July 1997 and February 2005. A 2-day protocol was used to localise the sentinel node with the injection of 99mTc-nanocolloid. There were two phases in the study: the learning phase (50 patients) and the application phase (741 patients). In the learning phase, a complete lymphadenectomy was always performed. In the application phase, sentinel nodes were studied postoperatively with breast cancer and lymphadenectomy was performed when considered warranted by the pathological postoperative results. The median follow-up duration in the 741 patients studied during the application phase was 32.3 months (range 6-72 months). In this phase a total of 787 sentinel nodes (719 axillary and 68 intramammary chain) were obtained (range 0-5 per patient, mean 1.01), with 153 (41 with micrometastasis) positive sentinel nodes. We observed a total of three FN SLN results (0.5%). All three presented as an axillary recurrence into 24 months from operation. After a median follow-up of 32.3 months we observed only three clinical recurrences among 741 patients. Our results indicate that the sentinel node protocol can give an adequate local control. PMID- 16244783 TI - Functional analysis of a novel RUNX2 missense mutation found in a family with cleidocranial dysplasia. AB - Mutations of the RUNX2 gene result in dominantly inherited cleidocranial dysplasia (CCD). RUNX2 encodes for an osteoblast-specific transcription factor, which recognizes specific DNA sequences by the runt domain. DNA binding is stabilized by the interaction with the protein CBFbeta, which induces structural modifications of the runt domain. A novel 574G > A RUNX2 missense mutation has been found in members of a family clinically diagnosed with CCD. This mutation causes the glycine at position 192 to change to arginine (G192R), in loop 9 of the runt domain. Unlike other residues of loop 9, G192 does not establish DNA contacts. Accordingly, the G192R mutant showed a 50% reduction in binding activity compared to the wild-type runt domain. However, the mutation completely abolished the activating properties of the protein on osteocalcin promoter. Moreover, the G192R mutant exerts a dominant-negative effect when overexpressed. Computer modeling indicated that the G192R mutation perturbs not only loop 9, but also other parts of the runt domain, suggesting impairment of the interaction with CBFbeta. PMID- 16244785 TI - The effect of race/ethnicity and insurance in the administration of standard therapy for local breast cancer in Florida. AB - OBJECTIVES: Assess the effect of race/ethnicity and insurance coverage on the receipt of standard treatment for local breast cancer. METHODS: Local breast cancers diagnosed between July 1997 and December 2000 and reported to Florida's registry were linked to the Agency of Healthcare Administration inpatient and outpatient databases, resulting in 23,817 female local breast cancers with informative treatment. Standard treatment was defined as mastectomy or breast conserving surgery followed by radiation therapy and it was modeled as a function of health insurance and race/ethnicity accounting for age at diagnosis, marital status and facility type. RESULTS: Approximately 88% of the local breast cancers received standard treatment. The likelihood of standard treatment decreased by 3% per year of increase in the age at diagnosis. Compared to white non-Hispanic, black non-Hispanic women were 19% less likely to receive standard treatment (OR=0.81, 95%CI=0.68, 0.97) and Hispanics were 23% less likely (OR=0.77, 95%CI=0.66, 0.89). Local breast cancers diagnosed in non-teaching facilities were 21% more likely to receive standard treatment compared to those diagnosed in teaching facilities (OR=1.21; 95%CI=1.05, 1.38)). Compared to single, married women were 51% more likely to get standard treatment (OR=1.51, 95%CI=1.31, 1.66), followed by separated or divorced women that were 37% more likely (OR=1.37, 95%CI =1.13, 1.66). Compared to the privately insured, Medicare beneficiaries were 36% more likely to receive standard treatment (OR=1.36, 95%CI=1.22, 1.51) whereas the uninsured were 24% less likely (OR=0.76, 95%CI=0.59, 0.96); Medicaid insured women were 29% less likely to receive standard treatment compared to the uninsured (OR=0.71, 95%CI=0.53, 0.96). CONCLUSION: Future efforts should target the elderly, Hispanic and black women, the uninsured, and those on Medicaid in order to reduce treatment disparities. PMID- 16244786 TI - A new germline mutation in BRCA1 gene in a sicilian family with ovarian cancer. AB - A group of 103 sicilian patients with hereditary and familiar breast and/or ovarian cancer were screened for Breast Cancer 1 gene (BRCA1) mutations by direct sequencing PCR products spanning the coding region and partial intronic regions of the BRCA1 gene. In this study, we report a new germline mutation in BRCA1 gene, not previously reported in the BIC database, in a woman with ovarian cancer at 46 years old. Mother's proband has been diagnosed the same histotype of ovarian cancer at 42 age. The mutational analyses that shown a 4843delC frameshift mutation in exon 16 of BRCA1 gene was extended to other family members including the proband's brother and her two sons. Direct automatic sequencing of DNA extracted from the lymphocytes showed exactly the same 4843delC frameshift mutation only in the brother. In conclusion, the characterization of this mutation could help in the identification of a founder mutation of sicilian area and this may provide significant advantages for genetic counselling. PMID- 16244787 TI - Intensity modulated radiation therapy (IMRT) reduces the dose to the contralateral breast when compared to conventional tangential fields for primary breast irradiation. AB - PURPOSE: To determine the dose received by the contralateral breast during primary breast irradiation using IMRT compared to conventional tangential field techniques. METHODS AND MATERIALS: Between March 2003 and March 2004, 83 patients with breast carcinoma were treated using 6, 10, or mixed 6/18 MV photons (65 with tangential IMRT technique and 18 with 3-dimensional technique using tangential fields with wedges) for primary breast irradiation following breast-conserving surgery. Paired thermoluminescent dosimeters (TLDs) were placed on each patient's contralateral breast, 4 and 8 cm from the center of the medial border of the tangential field. The TLDs were left on the patient during a single fraction and then measured 24 h afterwards. RESULTS: The mean dose delivered with photons to the primary breast for all patients was 4999 cGy (SD = 52) with a mean single fraction dose of 199 cGy (SD = 8). The mean percent of the prescribed dose to the contralateral breast measured at the 4- and 8-cm positions were 7.19% (SD = 2.28) and 4.63% (SD = 2.12), respectively, for patients treated with IMRT compared to 11.22% (SD = 2.73) and 10.70% (SD = 3.44), respectively, for the patients treated with conventional tangential field techniques. This represented a 36% and 57% reduction at the 4 and 8-cm contralateral positions, respectively, in the mean dose to the contralateral breast using IMRT compared to 3-D technique which was statistically significant (p < 0.0005, <0.0005, respectively). CONCLUSION: Primary breast irradiation with tangential IMRT technique significantly reduces the dose to the contralateral breast compared to conventional tangential field techniques. PMID- 16244788 TI - Breast tumors that overexpress nuclear metastasis-associated 1 (MTA1) protein have high recurrence risks but enhanced responses to systemic therapies. AB - Nuclear metastasis-associated 1(MTA1) protein is an estrogen receptor co repressor that regulates transcription via chromatin remodeling, and MTA1 messenger ribonucleic acid (mRNA) levels are elevated in several kinds of locally advanced and metastatic tumors relative to non-metastatic tumors. Previous studies in our laboratory mapped MTA1 into a region showing significantly lower LOH (loss of heterozygosity) in primary breast cancers with metastases compared to node-negative tumors, suggesting that epigenetic alterations of MTA1 affect metastatic potential. The present study examined immunohistochemical expression of the MTA1 protein in treated and untreated primary human breast cancers to study the relationship between MTA1 expression and clinical outcome. Node negative tumors that overexpress MTA1 protein had recurrence risks similar to node-positive tumors. In multivariate analysis of untreated node-negative tumors, highest expression of MTA1 was associated with increased relapse risk (hazard ratio (HR)=2.72, p=0.0003 for multivariate analysis). Tamoxifen and/or anthracylcene-based chemotherapies eliminated all MTA1 associations with clinical outcome, suggesting MTA1 overexpression predicts early disease relapse, but sensitizes breast tumors to systemic therapies. PMID- 16244789 TI - Melatonin enhances the inhibitory effect of aminoglutethimide on aromatase activity in MCF-7 human breast cancer cells. AB - The inhibition of the aromatase-induced intratumoral estrogen synthesis is one of the main anticancer pharmacological strategies. The aim of this paper was to study if a melatonin pretreatment prior to aminoglutethimide increases the efficiency of the aromatase inhibitor used in treating breast cancer. Aminoglutethimide (100 microM) and melatonin (1 nM) significantly decreased cellular aromatase activity in unpretreated MCF-7 cells. A sequential regimen of melatonin (1 nM) followed 24 h later by aminoglutethimide (100 microM) induced a significantly higher decrease in MCF-7 cell aromatase activity to below the values obtained in unpretreated cells. Melatonin treatment inhibited aromatase mRNA expression in unpretreated cells and a sequential treatment of cells with melatonin followed by aminoglutethimide induced a significant inhibition in the aromatase mRNA expression as compared to cells exposed to the same doses of aminoglutethimide, but without melatonin pretreatment. The present study demonstrates that a treatment with melatonin followed by aminoglutethimide is the most effective way of reducing the aromatase activity in the MCF-7 cell line. The aminoglutethimide inhibitory effect is more potent when MCF-7 cells are pre exposed to melatonin. Our results suggest that melatonin pretreatment increases the reduction of the aromatase activity of cells exposed to aminoglutethimide as a result of the decrease in the aromatase mRNA expression. The findings presented here point to melatonin pretreatment as a novel and interesting means to increase the efficacy of competitive aromatase inhibitors used in treating breast cancer. PMID- 16244790 TI - Overexpression of caveolin-1 and -2 in cell lines and in human samples of inflammatory breast cancer. AB - PURPOSE: Inflammatory breast cancer (IBC) is the most aggressive form of locally advanced breast cancer (LABC). The IBC phenotype is characterized by an infiltrative growth pattern, increased (lymph)angiogenesis and the propensity to invade dermal lymphatics. In pancreatic cancer, interactions between caveolin-1 and RhoC GTPase, a key molecule in causing the IBC phenotype, regulate tumour cell motility and invasion. In this study we sought to investigate the role of caveolin-1 and -2 in IBC cell lines and in human IBC samples. EXPERIMENTAL DESIGN: Differential methylation techniques identified the methylation status of the caveolin-1 and -2 promoters in human mammary epithelial cells (HMECs) and the SUM149 cell line. In cell line experiments, caveolin-1 and -2 mRNA and protein expression were compared in HMECs, MCF10A, the SUM102 non-IBC cell lines and 2 IBC cell lines (SUM149 and SUM190). Furthermore, caveolin-1 and -2 mRNA and protein expression were compared in human IBC and non-IBC samples using cDNA microarray, real-time qRT-PCR and immunohistochemistry. Results were correlated with RhoC protein expression data. RESULTS: In the SUM149 cell line, the caveolin 1 and -2 promoter sites were hypomethylated. A significantly increased expression of caveolin-1 and -2, both at the mRNA and protein level was found in IBC cell lines and in human samples of IBC: caveolin-1 and -2 mRNA were respectively 1.7 (p = 0.02) and 2.2 (p = 0.03) fold more expressed in IBC compared to non IBC and at the protein level, 41.4% of IBC specimens expressed either caveolin-1 or -2, compared to 15.6% of non-IBC specimens (p = 0.03). Furthermore a correlation was found between RhoC protein expression and caveolin-1 (p = 0.1) or caveolin-2 (p = 0.09) or either caveolin-1 or -2 protein expression (p = 0.04). CONCLUSIONS: Although considered a tumour suppressor in breast cancer, we demonstrated overexpression of caveolin-1 and -2 in IBC cell lines and in human samples of IBC, most likely due to hypomethylation of their respective promoters. These results confirm the distinct molecular signature of IBC. Our data further suggest interaction between RhoC GTPase and the caveolins in IBC. PMID- 16244791 TI - Endothelin B receptor agonist, IRL 1620, enhances the anti-tumor efficacy of paclitaxel in breast tumor rats. AB - Pharmacological agents that increase tumor blood flow could be utilized to promote the delivery of anti-cancer drugs. We have demonstrated that administration of endothelin-1 (ET-1) to breast tumor bearing rats transiently increased tumor blood flow by stimulating endothelin B (ET(B)) receptors. The present study evaluated the effect of ET(B) receptor agonist, IRL 1620, on breast tumor perfusion, concentration of [3H]paclitaxel in tumor and tissues, and efficacy of paclitaxel in N-methyl nitrosourea induced breast tumor bearing rats. Administration of IRL 1620 (3 and 9 nmol/kg) significantly increased (203 and 140%, respectively) breast tumor perfusion. BQ 788, an ET(B) receptor antagonist, pretreatment completely abolished IRL 1620 induced increase in tumor perfusion. Tumor [3H]paclitaxel concentration was increased by 308% when [3H]paclitaxel was administered 15 min after IRL 1620 (3 nmol/kg) compared to vehicle treated rats. However, IRL 1620 did not increase [3H]paclitaxel concentrations in other organs. Efficacy study showed that paclitaxel (5 mg/kg) administration on every third day for a total of five doses produced 60.0, 4.5 and 0% reduction in tumor volume, tumor progression and complete tumor remission, respectively, compared to saline treated rats. However, paclitaxel (5 mg/kg) when administered 15 min after IRL 1620 (3 nmol/kg) produced 268.9, 210.3 and 20% reduction in tumor volume, tumor progression and complete remission of tumors, respectively, compared to saline treated rats. In conclusion, IRL 1620 significantly enhanced delivery and effectiveness of paclitaxel in an animal model of breast cancer. PMID- 16244792 TI - Quantitative prediction of imprinting factor of molecularly imprinted polymers by artificial neural network. AB - Artificial neural network (ANN) implementing the back-propagation algorithm was applied for the calculation of the imprinting factors (IF) of molecularly imprinted polymers (MIP) as a function of the computed molecular descriptors of template and functional monomer molecules and mobile phase descriptors. The dataset used in our study were obtained from the literature and classified into two distinctive datasets on the basis of the polymer's morphology, irregularly sized MIP and uniformly sized MIP datasets. Results revealed that artificial neural network was able to perform well on datasets derived from uniformly sized MIP (n = 23, r = 0.946, RMS = 2.944) while performing poorly on datasets derived from irregularly sized MIP (n = 75, r = 0.382, RMS = 6.123). The superior performance of the uniformly sized MIP dataset over the irregularly sized MIP dataset could be attributed to its more predictable nature owing to the consistency of MIP particles, uniform number and association constant of binding sites, and minimal deviation of the imprinted polymers. The ability to predict the imprinting factor of imprinted polymer prior to performing actual experimental work provide great insights on the feasibility of the interaction between template-functional monomer pairs. PMID- 16244793 TI - Purification and characterization of curvaticin L442, a bacteriocin produced by Lactobacillus curvatus L442. AB - Lactobacillus curvatus L442, isolated from Greek traditional fermented sausage prepared without the addition of starters, produces a bacteriocin, curvaticin L442, which is active against the pathogen Listeria monocytogenes. The bacteriocin was purified by 50% ammonium sulphate precipitation, cation exchange, reverse phase and gel filtration chromatography. Partial N-terminal sequence analysis using Edman degradation revealed 30 amino acid residues, revealing high homology with the amino acid sequence of sakacin P. Curvaticin L442 is active at pH values between 4.0 and 9.0 and it retains activity even after incubation for 5 min at 121 degrees C with 1 atm of overpressure. Proteolytic enzymes and alpha amylase inactivated this curvaticin, while the effect of lipase was not severe. PMID- 16244794 TI - Acute monocular blindness resulting from transformation of von Recklinghausen's neurofibromatosis to malignant melanocytic schwannomas. AB - A 59-year-old male was transferred to our facility after initial workup for left eye blindness revealed multiple brain lesions. The patient presented with scant pedunculated skin lesions on his neck and arms, axillary freckling and bilateral subcutaneous ankle nodules suggestive of neurofibromatosis type 1 (NF-1). Neurological examination revealed left-sided blindness, diminished pupillary response to light, incomplete eyelid closure and facial droop. A CT series of the chest/abdomen/pelvis demonstrated several lesions in the lungs, liver and left kidney. CT-guided biopsy of the renal lesion was "consistent with metastatic melanoma" based on the presence of malignant cell staining for the melanocyte differentiation antigen MART-1, as well as the calcium binding protein S-100. A dermatology consultant confirmed neurofibromas and a cafe au lait macule. No dysplastic nevi or primary melanoma were identified. An ophthalmology consultant noted gray choroidal lesions on fundoscopic examination thought to be metastatic disease of unknown primary. Despite treatment with whole brain irradiation and intravenous steroids, the patient died on hospital day 10. A full autopsy showed diffuse neurofibromas and cafe au lait macules consistent with NF-1 and malignant melanocytic schwannomas with multiple metastases. PMID- 16244795 TI - LINAC radiosurgery as single treatment in cerebral metastases. AB - OBJECTIVE: Stereotactic radiosurgery is a radiation technique of high radiation dose focused on a stereotactic intracranial target in a single fraction with high precision. LINAC Radiosurgery has gained increasing relevance in the treatment of brain metastases since it was introduced by Sturm (1987). METHOD AND PATIENT SELECTION: From January 1996 to August 2003 110 patients were treated with LINAC Radiosurgery. A combination of the University of Florida system and the X Knife System developed by Radionics was used in all patients. Seventy patients had a single and 40 patients multiple metastatic lesions at the time of diagnosis and treatment. Overall 161 intracerebral metastases were treated. Median tumor volume was 3.1 ccm (0.3-15 ccm). Median radiation dose to the tumor margin was 1830 cGy (range 1100-2200 cGy) prescribed to the 80% isodose line. Whole brain radiation therapy with a total dose of 30 Gy in 10 fractions was performed in 35 patients because of multiple metastases and LINAC Radiosurgery was used as boost for recurrences. In 75 patients LINAC Radiosurgery was used as single treatment. RESULTS: The follow-up period was between 6 and 72 months. Local tumor control rate was 89.4%. Seventeen out of 161 metastases treated showed local recurrence. Eleven out of 75 patients treated with radiosurgery as single treatment developed distant recurrence and 3 out of 35 patients who were treated with whole brain radiation therapy (WBRT) and radiosurgery as boost. The 1-year survival rate is 54.9% with a median survival of 54 weeks. CONCLUSION: LINAC Radiosurgery is an effective and safe treatment modality in patients with cerebral metastases located in any area of the brain. WBRT should be preserved for patients with multiple metastases or be delayed until multiple recurrence occurs. Surgery is still the treatment of choice in metastases with mass effect and surgical accessible location. PMID- 16244796 TI - Evaluation of intraoperative autotransfusion filtration for hepatectomy and pancreatectomy. AB - BACKGROUND: Hepatectomy and pancreatectomy are often associated with significant intraoperative blood loss leading to postoperative anemia, which has been demonstrated to lead to increased perioperative morbidity, a prolonged hospital stay, and decreased overall survival. Cancer has remained an absolute contraindication to autotransfusion because of the unproven concern about reinfusion of malignant cells. Thus, the aim of this study was to test for the presence of malignant cells in autotransfused filtered blood in patients undergoing major pancreatic and liver resection. METHODS: A prospective study of 20 consecutive patients evaluated the presence of malignant cells from autotransfusion filtered blood after resection by flow cytometric and immunohistochemical methods. RESULTS: Ten patients underwent major hepatectomy for metastatic colorectal cancer, with a median blood loss of 500 mL (range, 200 700 mL). Three patients received a total of six units of packed red blood cells. Ten patients underwent pancreaticoduodenectomy for adenocarcinoma with a median blood loss of 400 mL (range, 200-1300 mL). Five patients received a total of nine units of packed red blood cells. Flow cytometry did not demonstrate the presence of any cytokeratin-positive carcinoma cells in filtered blood. CONCLUSIONS: Intraoperative autotransfusion for major hepatectomy in metastatic colorectal cancer and pancreatectomy for adenocarcinoma is safe and should begin to be evaluated in a phase II study for efficacy. PMID- 16244797 TI - Tumor regression and autoimmunity in cytotoxic T lymphocyte-associated antigen 4 blockade-treated patients. PMID- 16244798 TI - Limitations of ampullectomy in the treatment of nonfamilial ampullary neoplasms. AB - BACKGROUND: Pancreaticoduodenectomy (PD) is the standard surgical management of invasive ampullary neoplasms. A rational plan to use ampullectomy (AMP) for lesions at this location requires careful analysis of preoperative clinical information (comorbidity, lesion size, and histopathology) and intraoperative data (frozen section pathology and clinical impression) to properly select patients for this treatment. METHODS: We identified 140 consecutive cases of nonfamilial ampullary neoplasms from our prospective institutional database over a 7-year period (1996-2003). Preoperative and intraoperative factors were analyzed and related to outcomes. RESULTS: AMP was planned for 37 patients with small lesions (median, 1.86 cm [range, 0-3 cm] vs. 2.6 cm [range, 0-8 cm] in PD). AMP was converted to PD because of the extent of disease in three and an intraoperative diagnosis of invasive cancer in five patients. Preoperative biopsy had a diagnostic accuracy of 79% (97 of 123) but missed 23 cancers. Intraoperative frozen section had a diagnostic accuracy of 84%; two cases of high grade dysplasia and invasive cancer were missed. Patients with invasive cancer treated by AMP had a decreased recurrence-free and disease-specific survival compared with those treated by PD. Lymphatic spread of disease was associated with diminished long-term survival. Although both vascular invasion and tumor stage independently predicted lymphatic metastases, both were limited by their sensitivity. CONCLUSIONS: The reduced morbidity and mortality of AMP makes this the preferred treatment for benign lesions of the ampulla. Conversion to PD should be considered when intraoperative or final pathology identifies invasive adenocarcinoma. Refinement of clinicopathologic factors may reduce the occasional PD for benign disease and AMP for malignancy. PMID- 16244799 TI - Detection of carcinoembryonic antigen messenger RNA-expressing cells in portal and peripheral blood during surgery does not influence relapse in colorectal cancer. AB - BACKGROUND: No consensus has been reached on whether cancer cells detected in blood during colorectal cancer (CRC) surgery may serve as a prognostic indicator. METHODS: One hundred patients with CRC who underwent curative surgery were the subjects. Portal and peripheral blood were collected immediately after celiotomy and examined for carcinoembryonic antigen (CEA) messenger RNA (mRNA) by using competitive semi-nested reverse transcriptase-polymerase chain reaction. The median follow-up period was 59 months (range, 49-74 months). RESULTS: Until now, recurrence has been confirmed in 13 patients (13%). The 4-year recurrence rate was 6.7% (3 of 45) in patients with CEA mRNA-positive portal blood and 20.8% (10 of 48) in patients with CEA mRNA-negative portal blood (P = .09); it was 5.6% (2 of 36) and 19.3% (11 of 57) in patients with CEA mRNA-positive peripheral blood and CEA mRNA-negative blood, respectively (P = .12). There was no difference in disease-free survival between the CEA mRNA-positive and -negative groups. The multivariate analysis showed that the presence of tumor cells in portal or peripheral blood was a factor that reduced recurrence. The relative risks were .17 (P = .01) for the portal vein and .24 (P = .07) for the peripheral vein. CONCLUSIONS: The detection of cancer cells in blood taken during surgery is not considered to be a poor-prognostic factor in CRC. PMID- 16244800 TI - Evaluation of residual glandular tissue after skin-sparing mastectomies. AB - BACKGROUND: The oncological safety of skin-sparing mastectomy (SSM) has been the object of several studies. METHODS: From June 2003 to January 2004, 42 breast cancer patients, stage 0 to IIIA, underwent SSM. Before surgery, two lines were drawn on the breast skin, representing SSM and conventional mastectomy incisions. After surgery, the skin flap that would remain after SSM was removed, and immediate breast reconstruction was begun. The presence and amount of remaining glandular breast tissues were histologically evaluated in the skin flap. Terminal ductal lobular units (TDLUs) and residual disease were identified. These data were correlated with other clinical and pathologic parameters by using Fisher's exact test (P value) and receiver operating characteristic curves. RESULTS: The prevalence of residual breast tissue in the sample was 59.5%, and the presence of TDLUs was significantly associated with skin flaps thicker than 5 mm. Residual disease was found in 9.5% of the women and was associated with skin flaps >5 mm thick and the presence of TDLUs. There was no significant association between the presence of TDLUs and residual disease with age, body mass index, menopausal status, clinical and pathologic staging, breast volume, mammographic density, neoadjuvant chemotherapy, type of surgery, and presence of an extensive in situ component. The receiver operating characteristic curve showed that as skin flaps decrease in thickness, TDLUs also decrease. CONCLUSIONS: A high prevalence of glandular breast tissue and residual disease in the skin flap was associated with a skin flap thickness >5 mm. PMID- 16244801 TI - Inadequacy of lymph node staging in gastric cancer patients: a population-based study. AB - BACKGROUND: In 1997, examination of at least 15 lymph nodes was recommended for adequate gastric cancer staging. However, the proportion of patients undergoing an adequate lymph node examination (LNE) has not been studied in a population based manner. METHODS: We used Surveillance, Epidemiology, and End Results cancer registry data to assess LNE adequacy in adults with nonmetastatic gastric adenocarcinoma. We selected patients aged 18 through 80 years whose disease was diagnosed from 1998 through 2001 and who underwent at least partial gastrectomy. We evaluated the overall number of nodes, estimated the likelihood of adequate LNE (i.e., > or =15 nodes examined), and determined the influence of selected tumor and patient characteristics on LNE. RESULTS: In this 4-year period, 3593 patients met our study's selection criteria. The median number of nodes examined was 10: 32% of patients underwent adequate LNE, and 9% of patients had no nodes examined. Node-positive patients were more likely to have undergone an adequate LNE than node-negative patients (42% vs. 23%; P < .0001). Younger age, female sex, and more radical surgery were associated with adequate LNE in both univariate and multivariate analysis (P < .0001). Geographical site was an important predictor; patients from one registry (Hawaii) were significantly more likely to have undergone adequate LNE than patients from all other registries (56% vs. 30%; P < .0001). CONCLUSIONS: Our 4-year review of the Surveillance, Epidemiology, and End Results database revealed that only a third of patients with gastric cancer underwent adequate LNE, i.e., had the recommended minimum of 15 nodes examined for gastric cancer staging. Better results at one registry (Hawaii) indicate that substantial improvements could be made. PMID- 16244802 TI - Can the Memorial Sloan-Kettering Cancer Center nomogram predict the likelihood of nonsentinel lymph node metastases in breast cancer patients in the Netherlands? AB - BACKGROUND: According to Dutch guidelines, an axillary lymph node dissection (ALND) is recommended whenever a sentinel lymph node (SLN) contains metastatic disease. However, only in approximately 50% of patients with metastatic disease in the SLN are additional nodal metastases detected in the completion ALND. To identify the individual patient's risk for non-SLN metastases, a nomogram containing eight predictors was developed by the Breast Service of Memorial Sloan Kettering Cancer Center (New York, NY). The aim of this study was to test the accuracy of the nomogram on a population of Dutch breast cancer patients. METHODS: Patient, tumor, and SLN metastasis characteristics were collected for 222 consecutive patients who underwent a completion ALND. The data of the index and test populations were compared. A receiver operating characteristic curve was drawn, and the area under the curve was calculated to assess the discriminative power of the nomogram. RESULTS: Even though our patient population differed in many respects from the source population, the area under the receiver operating characteristic curve amounted to .77, a value very much comparable to the one found in the source population. CONCLUSIONS: The nomogram provides a fairly accurate predicted probability for the likelihood of non-SLN metastases in a general population of breast cancer patients at a regional teaching hospital in The Netherlands. This suggests that the nomogram's originally calculated predictive accuracy may be valid for patient populations that differ considerably from the population in which it was developed. PMID- 16244803 TI - Utility of breast magnetic resonance imaging in patients with occult primary breast cancer. AB - BACKGROUND: Although carcinoma presenting as axillary metastases is assumed to be due to breast cancer, identification of the primary lesion may prove problematic. We investigated the ability of breast magnetic resonance imaging (MRI) to identify the primary tumor, thereby confirming the diagnosis and broadening treatment options. METHODS: From 1995 to 2001, 69 patients at our institution presented with occult primary breast cancer. All patients had negative breast examinations and mammograms and underwent breast MRI. RESULTS: Of 69 patients, 55 had axillary adenopathy without evidence of distant disease (stage II); 14 had stage IV disease. In patients with stage II disease, MRI revealed suspicious lesions in 76% (42 of 55). In 62% (26 of 42), the MRI finding proved to be the occult primary tumor. Of these, 58% (15 of 26) were candidates for breast conservation. MRI did not identify the primary tumor in 25 women; 12 underwent mastectomy. Cancer was found in 33% (4 of 12) of these. Thirteen patients were treated with primary breast irradiation; three were lost to follow-up, one developed distant disease, and nine were without evidence of disease with a median follow-up of 4.5 years. In women with stage IV disease, MRI identified the primary tumor in 5 of 9 patients with regional adenopathy and 2 of 5 patients with distant disease (overall 50%; 7 of 14). MRI identified the primary tumor in women with both mammographically dense (19 of 44; 43%) and less dense (10 of 20; 50%) breasts. CONCLUSIONS: Breast MRI detects mammographically occult cancer in half of women with axillary metastases, regardless of breast density. MRI is a powerful tool for stage II and stage IV patients with occult primary breast cancer. PMID- 16244804 TI - Minimally invasive surgery for colorectal carcinoma. PMID- 16244805 TI - Effectiveness of palliative procedures for intra-abdominal sarcomas. AB - BACKGROUND: Nearly half of patients with intra-abdominal (retroperitoneal, visceral, or pelvic) sarcoma undergo more than one operation. When the objective shifts from cure to palliation, the clinical quandary of doing no harm and maximizing benefit is magnified. Knowledge of the effectiveness of a procedure at achieving its palliative intent, as well as its attendant morbidity and mortality, is therefore paramount during this deliberation. METHODS: A retrospective review was performed of all patients with a diagnosis of intra abdominal sarcoma who underwent a palliative procedure between 1982 and 2003. A procedure was defined as palliative if it was explicitly performed to relieve symptoms. RESULTS: Ten percent (112 of 1084) of patients with a diagnosis of intra-abdominal sarcoma underwent a total of 156 palliative procedures. The most frequent system for which a palliative procedure was performed was gastrointestinal (68 of 156; 44%). Overall, 71% of patients had improvement of symptoms 30 days after the operation, whereas only 54% of patients remained symptom free after 100 days. Although 54% of gastrointestinal tract obstructive symptoms were successfully relieved at 30 days, only 23% of patients remained symptom free at 100 days. The overall operative morbidity was 29%, and postoperative mortality was 12%. Patients undergoing procedures intended to palliate gastrointestinal obstruction encountered the greatest morbidity (19 of 40; 48%). CONCLUSIONS: Successful palliation of many symptoms associated with advanced intra-abdominal sarcoma may be achieved. However, even in highly selected patients, the progressive and pervasive nature of the disease limits the opportunity to attain sustained relief for gastrointestinal obstructive symptoms. PMID- 16244806 TI - Herpes simplex virus amplicon delivery of a hypoxia-inducible soluble vascular endothelial growth factor receptor (sFlk-1) inhibits angiogenesis and tumor growth in pancreatic adenocarcinoma. AB - BACKGROUND: Tumor hypoxia induces vascular endothelial growth factor (VEGF) expression, which stimulates angiogenesis and tumor proliferation. The VEGF signaling pathway is inhibited by soluble VEGF receptors (soluble fetal liver kinase 1; sFlk-1), which bind VEGF and block its interaction with endothelial cells. Herpes simplex virus (HSV) amplicons are replication-incompetent viruses used for gene delivery. We attempted to attenuate angiogenesis and inhibit pancreatic tumor growth through HSV amplicon-mediated expression of sFlk-1 under hypoxic control. METHODS: A multimerized hypoxia-responsive enhancer (10 x HRE) was cloned upstream of the sFlk-1 gene (10 x HRE/sFlk-1). A novel HSV amplicon expressing 10 x HRE/sFlk-1 was genetically engineered (HSV10 x HRE/sFlk-1).Human pancreatic adenocarcinoma cells (AsPC1) were transduced with HSV10 x HRE/sFlk-1 and incubated in normoxia (21% oxygen) or hypoxia (1% oxygen). Capillary inhibition was evaluated by human umbilical vein endothelial cell assay. Western blot assessed sFlk-1 expression. AsPC1 flank tumor xenografts (n = 24) were transduced with HSV10 x HRE/sFlk-1. RESULTS: Media from normoxic AsPC1 transduced with HSV10 x HRE/sFlk-1 yielded a 36% reduction in capillary formation versus controls (P < .05), whereas hypoxic AsPC1 yielded a 76% reduction (P < .005). Western blot of AsPC1 transduced with HSV10 x HRE/sFlk-1 demonstrated greater sFlk-1 expression in hypoxia versus normoxia. AsPC1 flank tumors treated with HSV10 x HRE/sFlk-1 exhibited a 59% reduction in volume versus controls (P < .000001). CONCLUSIONS: HSV amplicon delivery of a hypoxia-inducible soluble VEGF receptor significantly reduces new vessel formation and tumor growth. Tumor hypoxia can thus be used to direct antiangiogenic therapy to pancreatic adenocarcinoma. PMID- 16244807 TI - Nuclear DNA contents, rDNAs, and karyotype evolution in Vicia subgenus Vicia: II. Section Peregrinae. AB - Nuclear DNA contents, automated karyotype analyses, and sequences of rDNA spacers have been determined for the species of Vicia belonging to sect. Peregrinae, as well as for V. mollis. The phylogenetic data generated from the comparison of rDNA sequences and karyomorphological results would both indicate that Vicia mollis is a sister group to sect. Peregrinae. The relationships among the species belonging to the Peregrinae section and species enclosed in sections Faba, Narbonensis, and Bithynicae have been also investigated: a clade including V. mollis and sect. Peregrinae is a sister group to a clade including V. bithynica and sect. Narbonensis. With our choice of outgroup, Vicia faba (including subsp. paucijuga) is external to the above mentioned inclusive group. PMID- 16244808 TI - Superoxide dismutase, peroxidase, and germin-like protein activity in plasma membranes and apoplast of maize roots. AB - The analysis of plasma membranes from maize roots by native gel electrophoresis revealed the existence of Mn-containing 120 kDa and CuZn-containing 70, 40, and 15 kDa superoxide dismutase (SOD) isoform activities. Isoelectric focusing of the plasma membranes differentiated anionic SOD isoforms with a pI of about 5 and cationic SOD isoforms at pI 8.6. Solubilization of the plasma membrane proteins further separated the cationic SOD into pI 8.6, 8.2, 8.4, and 7.2 isoforms. Double staining for both SOD and peroxidase activities showed an overlap of these activities only in the case of the high-molecular-mass (ca. 120 kDa) isoforms. High-temperature treatments demonstrated that the 120 kDa isoform was active even at 100 degrees C, indicating that it was a germin-like protein with superoxide dismutating activity, different from the peroxidase with a similar molecular mass and the lower-molecular-mass CuZn-containing superoxide dismutases. These results are compared to those obtained from whole-tissue extract and apoplastic fluid. PMID- 16244810 TI - Extra thin alginate film: an efficient technique for protoplast culture. AB - This paper reports an efficient protoplast culture technique, the "extra thin alginate film" technique. The development of this improved method of protoplast culture was an outcome of an assessment of the efficiency and shortcomings of various protoplast culture techniques. The efficiency of this technique was evaluated with two model plant systems, viz., Nicotiana tabacum and Lotus corniculatus, and a comparison was made with the "thin alginate layer" technique, another efficient protoplast culture system. Results indicate that the culture technique with extra thin alginate film is as efficient as the technique with thin alginate layer, with many additional advantages. The present innovation overcomes most of the limitations of protoplast culture techniques described so far and can now be applied to a wide variety of crops to check its general applicability. PMID- 16244809 TI - Poly(A)+ RNA and cytoskeleton during cyst formation in the cap ray of Acetabularia peniculus. AB - The configuration and distribution of polyadenylated RNA (poly(A)+ RNA) during cyst formation in the cap rays of Acetabularia peniculus were demonstrated by fluorescence in situ hybridization using oligo(dT) as a probe, and the spatial and functional relationships between poly(A)+ RNA and microtubules or actin filaments were examined by immunofluorescence microscopy and cytoskeletal inhibitor treatment. Poly(A)+ RNA striations were present in the cytoplasm of early cap rays and associated with longitudinal actin bundles. Cytochalasin D destroyed the actin filaments and caused a dispersal of the striations. Poly(A)+ RNA striations occurred in the cytoplasm of the cap rays up to the stage when secondary nuclei migrated into the cap rays, but they disappeared after the secondary nuclei were settled in their positions. At that time, a mass of poly(A)+ RNA was present around each of the secondary nuclei and accumulated rRNA. This mass colocalized with microtubules radiating from the surface of each secondary nucleus and disappeared when the microtubules were depolymerized by butamifos, which did not affect the configuration of actin filaments. These masses of poly(A)+ RNA continued to exist even after the cap ray cytoplasm divided into cyst domains. Thus two distinct forms of poly(A)+ RNA population, striations and masses, appear in turn at consecutive stages of cyst formation and are associated with distinct cytoskeletal elements, actin filaments and microtubules, respectively. PMID- 16244812 TI - Re-designing state mental health policy to prevent the use of seclusion and restraint. AB - : The members of the National Association of State Mental Health Program Directors (NASMHPD) believe that seclusion and restraint, including "chemical restraints," are safety interventions of last resort and are not treatment interventions. Seclusion and restraint should never be used for the purposes of discipline, coercion, or staff convenience, or as a replacement for adequate levels of staff or active treatment. The use of seclusion and restraint creates significant risks for people with psychiatric disabilities. These risks include serious injury or death, retraumatization of people who have a history of trauma, and loss of dignity and other psychological harm. In light of these potential serious consequences, seclusion and restraint should be used only when there exists an imminent risk of danger to the individual or others and no other safe and effective intervention is possible. (Endorsed by the State Mental Health Directors, July 13, 1999). (NASMHPD 1999, NASMHPD Position Statement on Seclusion and Restraint. Alexandria, VA: National Technical Assistance Center for State Mental Health Planning.). PMID- 16244811 TI - Role of dislocation loops on the elastic constants of lyotropic lamellar phases. AB - We study the role of dislocation loops defects on the elasticity of lamellar phases by investigating the variation of the lamellar elastic constants, B and K, induced by the proliferation of these defects. We focus our interest on one particular lamellar phase made up of a mixture of C(12)E(5) and DMPC in water, which is already well-characterised. This lamellar phase undergoes a second-order (or weakly first-order) lamellar-to-nematic phase transition at about 19 degrees C and dislocation loops are seen to proliferate within the lamellar structure when temperature is decreased below 30 degrees C. The values of both elastic constants of this given lamellar phase are measured as a function of temperature, approaching the lamellar-to-nematic transition, with the help of Quasi-Elastic Light Scattering (QELS) on oriented lamellar phases. Very surprisingly we observe a strong and rapid increase in both Band K as the lamellar-to-nematic transition temperature is approached. These increases are seen to start as soon as dislocation loops can be observed in the lamellar phase. We interpret our results as being the consequence of the appearance and proliferation of dislocation loops within the lamellar structure. According to a simple model we developed we show that B and K are proportional to the density of dislocation loops in the lamellar phase. PMID- 16244814 TI - [Nutrition for the elderly]. PMID- 16244815 TI - [Bioelectric impedance analysis in the diagnosis of malnutrition]. AB - Malnutrition is one of the most common comorbidities in the elderly. Despite several screening and assessment tools, the diagnosis can be difficult in some cases. In the geriatric population, classical anthropometry does not help very much. For that reason there is a need for further diagnostic criteria. Bioelectrical impedance analysis (BIA) is a fast, low-cost and non-invasive method, which has shown its value in several fields of medicine. Whether this method could be a useful part in the diagnosis of malnutrition in geriatric patients, is the question of this study. In comparison with the Mini-Nutritional Assessment (MNA), serum-albumin and other diagnostic markers of malnutrition, especially the phase angle (50 kHz) of bioelectric impedance analysis showed a consistent correlation with most of the parameters. The calculation of body-fat mass and fat-free mass from bioelectrical impedance analysis is not yet sufficiently validated for geriatric patients. In this study there was no good correlation of the BIA-calculated body-fat mass and fat-free mass with the observed nutritional parameters. PMID- 16244813 TI - Motor dysfunction in patients with liver cirrhosis: impairment of handwriting. AB - Motor dysfunction is an important clinical finding in patients with liver cirrhosis and mild forms of hepatic encephalopathy. The mechanisms and clinical appearance of motor impairment in patients with liver cirrhosis are not completely understood. We studied fine motor control in forty four patients with advanced liver cirrhosis (excluding those with hepatic encephalopathy grade II) and 48 healthy controls using a kinematic analysis of standardized handwriting tests. We analysed parameters of velocity, the ability to coordinate and the level of automatisation of handwriting movements. Furthermore, we studied the association between impairment of handwriting and clinical neuro-psychiatric symptoms. As compared with control subjects, patients showed a statistically significant reduction of movement peak velocity in all handwriting tasks as well as a substantial increase of number of velocity inversions per stroke. Using a z score based assessment we found impairment of handwriting in fourteen out of forty four patients (31.8 %). The deterioration of handwriting was associated with clinical symptoms of motor dysfunction, such as bradykinesia, adiadochokinesia, dysmetria of upper extremities and gait ataxia. This is the first study that quantitatively investigates impairment of handwriting in patients with liver cirrhosis. Our findings suggest the application of kinematic analysis of handwriting for diagnostics of motor dysfunction in patients with mild forms of hepatic encephalopathy. PMID- 16244816 TI - Comparison of the Mini Nutritional Assessment, Subjective Global Assessment, and Nutritional Risk Screening (NRS 2002) for nutritional screening and assessment in geriatric hospital patients. AB - The Mini Nutritional Assessment (MNA), the Subjective Global Assessment (SGA) and the Nutritional Risk Screening (NRS) are screening and assessment tools aimed at detecting malnourished individuals and those at risk for malnutrition. In our study we tested their applicability in geriatric hospital patients and compared the results of the three tools. We examined prospectively all patients of two acute geriatric wards by the MNA, the SGA and the NRS. 121 patients were included in the study. The MNA could be completed in 66.1% of all patients, the SGA in 99.2% and the NRS in 98.3%. There was a significant association of all three tools with the BMI (p<0.01). With regard to serum albumin and to length of hospital stay (p<0.05), only a significant association could be shown for the MNA (p<0.05). Although the categories of the results were not completely identical for the three tools there were more patients at risk or malnourished according to the MNA (70%) than according to the SGA (45%) or the NRS (40.3%). The direct comparison of the NRS with the MNA and the SGA demonstrated significant differences, especially for the latter (p<0.001). In a relevant percentage of those tested, MNA, SGA, and NRS identify different individuals as malnourished or at risk for malnutrition. Because of its association with relevant prognostic parameters, the MNA is still the first choice for geriatric hospital patients. For those patients to whom the MNA cannot be applied, the NRS is recommended. PMID- 16244817 TI - [Healthy seniors with a normal nutritional level in the Mini-Nutritional Assessment (MNA) identified as at risk for status decline and impaired function]. AB - Older adults are potentially vulnerable for malnutrition, risk which can be assessed by the Mini-Nutritional Assessment (MNA). To answer the question, whether the MNA can also assess healthy, normal nourished seniors and to check for correlations on functional disabilities and risk factors for functional status decline, 58 independent living, healthy elderly persons with an MNA above 24 points were studied. Different clinical and laboratory exams were performed and a comprehensive geriatric assessment was carried out. There was a non age dependent correlation for the number of risks in the Lachs' screening, for the number of drugs taken, for the hemoglobin concentration, the level of the g- GT and for mobility assessment tested by the "Timed Up and Go Test" and the "POMA". The MNA can also distinguish in the "normal" range from 24 to 30 points between persons at risk for functional status decline or persons with slightly impaired functions. PMID- 16244818 TI - [Oral health, dental state and nutrition in older adults]. AB - The loss of natural teeth impairs essentially the chewing function and can only partly be restored by the insertion of dental prostheses. Equally, xerostomia and dysphagia may aggravate the nutritional intake in older adults. Often denture wearers do subjectively not notice the adjustment of their food choice and the employment of special preparation techniques. Finally the dental state influences the nutritional intake. A reduced number of teeth correlates with the intake of less calories, proteins, fat, non-starch polysaccharides and vitamins. Often missing calories are compensated by an increased consumption of sugar and fat. Especially edentulous persons with a low level of education choose a diet which is rich in fat and sugar. Further the daily intake of fruit and vegetables diminishes along with fewer occlusal contacts in posterior teeth. The restoration of the chewing function by dental intervention does not lead to an improvement of the nutritional intake by itself and should therefore always be complemented by nutritional advice. PMID- 16244819 TI - [Oral health-related quality of life in elderly]. AB - PURPOSE: The aim of this study was to describe the oral health related quality of life (OHRQoL) of institutionalized elderly. MATERIAL: 159 elderly were randomly selected for this, study ranging in age from 61 to 98 (mean age 82.8; 19.1% male, 80.9% female). For measuring the OHRQoL the Oral Health Impact Profile (OHIP) was used. Additionally, a dental status was evaluated. RESULTS: 15% of the subjects had only natural teeth or fixed dentures, 80% removable dentures and 15% no dentures at all. OHIP-mean reached 33+/-25 points, between 0 and 112. The subcategory "functional limitation" showed the highest impairment (6.3%). The items with the highest impairment on OHRQoL in the different subcategories were: dentures not fitting properly, uncomfortable dentures, self-conscious due to teeth, avoid eating some foods, felt depressed, avoid going out and general health worsened. CONCLUSIONS: The OHIP summary score represents an overview of OHRQoL and answers detailed questions with the help of the single items. In institutionalized elderly many items showed impairment, especially in items concerning the prosthetic restorations. This demonstrates the need for dental aftercare and not only dental interventions for acute pain. Establishing aftercare and prophylaxis could increase the OHRQoL and subsequently the overall well-being. PMID- 16244820 TI - [Maillard reaction products in food as pro-inflammatory and pro-arteriosclerotic factors of degenerative diseases]. AB - Heating of food induces the formation of Maillard reaction products (MRPs) caused by the reaction of reducing sugars with proteins or amino acids. Analogous reactions occur in the human body, eventually forming "Advanced Glycation Endproducts" (AGEs). AGEs accumulate in aging tissues accelerating degenerative inflammatory and proliferative processes. MRPs present in food can also directly cause inflammatory processes in the intestines and, once absorbed, would support and reinforce any inflammatory and degenerative process occurring in the body. The contribution of AGEs (and additional MRPs) in the development of diabetic complications as well as nephropathy, neuropathy, micro- and macroangiopathies is now well established. Which of the MRPs or AGEs in particular induce these cellular processes is currently unknown. Thus the exact knowledge of the chemical structures of the MRPs could help to minimize the formation of "harmful MRPs" that occur due to heating in food processing. Because MRPs play a decisive role in the successful marketing of edibles due to their characteristics as flavor components, it is important to increase the amount of innocuous and palatable MRPs, and minimize signal active pro-inflammatory MRPs by the use of defined preparation methods. It is practicable to use low-priced immunological methods for the quantitative determination of specific MRPs or AGEs. In the medical area, the knowledge of the signal active MRP/AGE structures provides the opportunity to measure their concentrations in body fluids and tissues and thus determine their influence on inflammatory and age-related degenerative processes (e. g., late diabetic complications, arteriosclerosis, degeneration of neurons). From a clinical perspective, the application of RAGE antagonists after an appropriate chemical diagnosis could be effective in supporting the treatment of affected patient groups, especially older diabetic and dialysis patients. PMID- 16244821 TI - [What does the language of Alzheimer patients have to do with the language of Paul Celan?]. AB - Being in touch with severely demented patients requires a fundamental transformation in communication habits in medicine and in nursing. There is a need of reorientation away from an interpreting and often deficit-oriented aspect of communication towards attentive observance of unusual verbal and nonverbal signals and resources of the patient. Spontaneous and open communication with the patient outweighs the importance of a goal-oriented exchange of information and poses a significant challenge for all health and social professions. The well being of persons with dementia depends mainly on the quality of communication and on the design of the milieu and the quality of everyday life. Interaction in nursing seems to be the crucial issue, including both spontaneity and creativity in the interaction partners, while respecting the personal boundaries of both the patient and the nurse in the necessary intimacy of the care environment. This essay shows important aspects and strategies of adequate communication with people with dementia from the perspective of medicine and of nursing. Ways to improve communication skills are shown, referring, among others, to the approaches by Kate Allan and John Killick (research fellows at Dementia Services Development Centre, University of Stirling). PMID- 16244822 TI - Assessment of cognitive status in the elderly using telephone interviews. AB - OBJECTIVE: To examine the feasibility to assess cognitive status in the elderly using telephone interviews. RESEARCH DESIGN AND METHODS: From January to December 2003, 740 participants of the ongoing Heidelberg longitudinal study (HeiDE) aged 70 years or more were eligible for a telephone interview on cognitive status. Validated instruments to assess cognitive status, including the Telephone Interview of Cognitive Status (TICS) and the East Boston Memory Test (EBMT, immediate and delayed recall), the Verbal Fluency Test, the HAWIE-"Vocabulary Test" and the HAWIE-"General Knowledge", a prospective memory test, and a digit span backwards-test were translated into German, if applicable. RESULTS: Out of 740 participants at the age of 70 or older, 473 participants were interviewed (64.9%). The total score of the TICS (maximum=best: 41) ranged from 21 to 40 (mean 33.5, SD 3.1; median 34.0). The EBMT scores (immediate recall; maximum=best: 12) ranged from 4 to 12 (mean 9.2, SD 1.7; median 9.0). CONCLUSIONS: Cognitive status could be successfully assessed by telephone interview in elderly participants of an ongoing population-based cohort study. Specifically, some of the tests showed pronounced variability allowing cross sectional analyses whereas others seem more valuable for longitudinal assessment. PMID- 16244826 TI - [Nonoperative therapy for arthritis]. PMID- 16244827 TI - [Health education and health training with osteoarthritis]. AB - Osteoarthrosis is one of the most prevalent diseases in orthopedics. Its impact for the individual patient as well as for the socio-economic system is enormous. An increasing loss of function and pain progress of disease leads to chronic handicap, which severely restricts freedom of a self-determined life and reduces quality of life. Besides conservative and later surgical treatment, health inducing offers to patients with arthrosis deformans have gained more importance in the process of coping with the illness similar to other chronic diseases. Health education and exercise are supposed to help patients gain more theoretical and practical knowledge of the disease and foster a more positive attitude in order to improve self management. The effect of patient educating programs with respect to self-help activities and socio-economic savings (return to work, days of hospital treatment) could be verified during recent years in many cases of chronic disease, such as rheumatic disease, diabetes and asthma through prospective multicenter studies. Whether special health training in osteoarthrosis leads to similar effects is subject of current studies. PMID- 16244828 TI - [Physical therapy of osteoarthritis]. AB - Physical therapy is part of guidelines and recommendations in the treatment of osteoarthritis. Different methods of physical therapy are used in osteoarthritis. There is evidence that manual physical therapy and exercise improve function and reduce pain in osteoarthritic joints. Thermal modalities are employed for short term pain relief and change the intraarticular temperature. Electrotherapy, therapeutic ultrasound and balneotherapy show positive therapeutic effects. Based on studies and clinical experience, physical therapy must be recommended in the therapy of osteoarthritis. PMID- 16244829 TI - [Evidence-based evaluation of study results of symptomatic glucosamine therapy]. AB - Meta-analyses can contribute to the field of evidence based medicine. In the systematic analyses so far published, the symptomatic efficacy of glucosamine was described to be moderate to large. The results of the individual clinical studies however do not support these findings unanimously. In the past couple of years, some new studies of high quality standard were published. This publication gives an overview and an evaluation of the current available studies for the symptomatic treatment of patients with gonarthrosis with glucosamine. It also compares the results of the individual studies with those of recent meta analyses. PMID- 16244830 TI - [Pharmacological treatment of osteoarthritis in the elderly]. AB - Osteoarthritis (OA) is the most common cause of functional disability in the elderly. Pain and loss of motion induce a vicious circle, leading to instability, frailty and ultimately invalidity. Currently, there is no treatment to reverse or slow the disease progression to a clinically meaningful extent. Thus, the primary goal of OA treatment in the elderly is pain relief and preservation of joint function. For this, pharmacological, non-pharmacological and if necessary surgical treatment regimes must form an integrated concept. However, the real challenge is polymorbidity and other age-related or age-associated factors, which influence the course of disease and its therapy unfavorably. The changes in pharmacokinetics and -dynamics in the elderly can be compensated for the nonopioid and opioid-analgesics by the well known "start low, go slow" approach. More problematic are non-steroidal anti-inflammatory drugs (NSAIDs), which are most often used for symptomatic treatment of OA: Patients over 65 have an enhanced susceptibility to the gastrointestinal and renal side effects of NSAIDs; all NSAIDs, not only coxibs, increase the cardiovascular risk in patients with such a disease; number and severity of drug interactions is elevated due to age associated polypharmacy. Thus, NSAIDs, including coxibs, should be used with great caution for treatment of OA in the elderly. PMID- 16244831 TI - [Imaging techniques in rheumatology: conventional radiography in rheumatoid arthritis]. AB - Conventional radiography is still regarded the gold standard for imaging techniques in rheumatoid arthritis (RA). It is a very important tool for the diagnosis, the differential diagnosis, and the evaluation of the course and prognosis of the disease. Special advantages of conventional radiography are the worldwide availability and experience with the method over decades and the relatively low costs. Moreover, x-rays can be stored and re-evaluated over long periods of time. Joint damage caused by RA can be quantified by means of scoring methods. The amount of destruction correlates well with functional disability over time. The inhibition of damage progression seen on radiographs is the most important characteristic of a DMARD. Typical radiographic changes are part of the ACR classification criteria of RA. Technically, bone structure can be demonstrated with a high local resolution better than with all other imaging techniques, whereas only indirect conclusions can be drawn concerning soft tissue and cartilage lesions. This review includes recommendations given by the "Commission on Imaging Techniques" of the German Society of Rheumatology regarding technical and personal preconditions, costs, indication as well as practical performance of radiography in RA. In addition, radiographic changes that can be expected in RA including destruction and repair are discussed briefly. PMID- 16244833 TI - [PM-Scl antibody positive systemic sclerosis associated with inclusion-body myositis]. AB - We describe a 72- year-old patient with a ten year history of anti-PM-Scl positive systemic sclerosis associated with inclusion-body myositis. While the association of dermatomyositis and polymyositis with anti-PM-Scl positive systemic sclerosis is frequently reported, inclusion-body myositis was, to the best of our knowledge, only previously described once in association with anti-PM Scl-positive systemic sclerosis. The distinction between inclusion-body myositis and other forms of inflammatory myopathy, like the histopathologically well distinguishable polymyositis or dermatomyositis, is relevant because of the poor response of inclusion- body myositis to immunosuppressive treatment. Our case underlines that in patients with anti-PM-Scl-positive systemic sclerosis and treatment resistant progressive myopathy the diagnosis of inclusion body myositis should be considered. PMID- 16244832 TI - [The common occurrence of osteoarthritis and osteoporosis and the value of markers of bone turnover]. AB - OBJECTIVE: Increasing bone mineral density (BMD) has been found in several studies in patients with osteoarthritis. Therefore, many clinicians deny the simultaneous occurrence of osteoporosis (OP) and osteoarthritis (OA). Because of our clinical impression however, we suggested that we have to consider a common occurrence. Furthermore, the value of markers of bone turn over with a view to early diagnosis of OP and or as an assessment for bone metabolism in OA is still a matter debate and their clinical use has not been clearly defined in the management of the individual patient. METHOD: The BMD of the lumbar spine and the proximal femur of 119 OA patients (83 postmenopausal female patients aged 50-83 and 35 male patients aged 36-86 years) who subsequently required hip or knee replacements, but were otherwise healthy, were measured by dual energy X-ray absorption (DXA), Hologic QDR-2000. We also measured biochemical markers of bone turn over, i. e., CICP, ICTP, DPD, PTH, estrogen, testosterone, bAP, hydroxy vitamin D and the normal blood count. RESULTS: There was a high occurrence of a low BMD among the patients. A total of 28.9% of women were affected by OP and 52.9% by osteopoenie. This reflects the normal distribution of OP in the female population. Of the male patients 20% had OP and 38.8% osteopoenie. This is astonishing high. Age proved to be a significant factor in the degree of BMD. An association between disuse osteoporosis and degree of BMD in the OA affected joint could not be proven. The use of the biochemical markers for an earlier diagnosis or to assess bone metabolism in OP and OA was not possible. CONCLUSION: We can not support the hypotheses that OA prevents OP. Moreover, the occurrence of OP in our study reflected the incidence of OP in the average female and the astonishingly high incidence in the male population; however does not mean that the two conditions are mutually exclusive. We did not find that the biochemical markers of bone turn over could deliver additional information with respect to bone metabolism and an earlier diagnosis of OP. PMID- 16244838 TI - [Granuloma annulare]. AB - Granuloma annulare is a benign self-limiting disease of unknown origin, characterized by dermal papules tending to form annular shapes. It belongs to the granulomatous inflammatory skin diseases characterized by histomorphologically detectable granulomas. In addition to localized variants, generalized or disseminated variants also exist. The disease is often asymptomatic and does not require therapy in every case. Established therapies for localized forms of granuloma annulare are glucocorticosteroids, cream PUVA and cryosurgery. Antimalarials, antileprotics, retinoids, fumarates, UVA1 phototherapy and photochemotherapy are of value in disseminated cases. Recent case reports demonstrate the successful treatment of granuloma annulare with topical calcineurin antagonists or imiquimod and with the systemic administration of biologicals. PMID- 16244839 TI - Some notes on medical English. AB - Writing medical papers is a challenge in any language, but writing in a foreign language such as English can be especially difficult. This paper provides some suggestions on how to overcome common problems in preparing medical papers and presentations, for writers whose native language isn't English. Stylistic and grammatical peculiarities of the English language are considered, as well as Americanizations of Latin terminology, scientific ("international") notation, variations in usage between Britain and the U.S.A., scientific and ethical standards of medical publications in English, and some resources for foreign language researchers who want to compose papers or presentations in English. PMID- 16244840 TI - A comparison of graphical and textual presentations of time series data to support medical decision making in the neonatal intensive care unit. AB - OBJECTIVE: To compare expert-generated textual summaries of physiological data with trend graphs, in terms of their ability to support neonatal Intensive Care Unit (ICU) staff in making decisions when presented with medical scenarios. METHODS: Forty neonatal ICU staff were recruited for the experiment, eight from each of five groups--junior, intermediate and senior nurses, junior and senior doctors. The participants were presented with medical scenarios on a computer screen, and asked to choose from a list of 18 possible actions those they thought were appropriate. Half of the scenarios were presented as trend graphs, while the other half were presented as passages of text. The textual summaries had been generated by two human experts and were intended to describe the physiological state of the patient over a short period of time (around 40 minutes) but not to interpret it. RESULTS: In terms of the content of responses there was a clear advantage for the Text condition, with participants tending to choose more of the appropriate actions when the information was presented as text rather than as graphs. In terms of the speed of response there was no difference between the Graphs and Text conditions. There was no significant difference between the staff groups in terms of speed or content of responses. In contrast to the objective measures of performance, the majority of participants reported a subjective preference for the Graphs condition. CONCLUSIONS: In this experimental task, participants performed better when presented with a textual summary of the medical scenario than when it was presented as a set of trend graphs. If the necessary algorithms could be developed that would allow computers automatically to generate descriptive summaries of physiological data, this could potentially be a useful feature of decision support tools in the intensive care unit. PMID- 16244841 TI - Reliability of near-infrared spectroscopy in people with dark skin pigmentation. AB - OBJECTIVE: Near-infrared spectroscopy (NIRS) is a promising non-invasive technique for the continuous monitoring of tissue oxygen delivery. NIRS detects light absorbance of haemoglobin chromophores to determine tissue oxygen saturation (StO2). As skin colour is also determined by the presence of chromophores, it is plausible that NIRS signal quality may be affected by dark skin pigmentation. METHODS: Tissue saturation in the anterior compartment of the lower leg during isometric contraction was measured using NIRS in 17 volunteers with dark skin pigmentation. Measurements were continued until StO2 was zero percent or until the signal disappeared. RESULTS: The NIRS device failed to register tissue saturation values at some point in nine of seventeen volunteers. This occurred more often in individuals with darker skin. CONCLUSIONS: In patients with a dark pigmented skin, NIRS StO2 measurements should be interpreted with caution, as melanin clearly interferes with the quality of the reflected NIRS signal. PMID- 16244843 TI - ECG compression by modelling the instantaneous module/phase of its DCT. AB - Recent developments in compression methods on the non-linear and non-stationary data, such as electrocardiograms (ECG), have received large attention by the time frequency analysts. The technique presented in this paper is based on parametrical modeling the instantaneous module as well as the instantaneous phase, estimated directly from the Discrete Cosine Transform (DCT) of each ECG beat. The estimated parameters are then used to reconstruct each recorded beat. In order to evaluate the performance of our technique, data recorded from the MIT BIH arrhythmia database are used. PMID- 16244842 TI - Recovery from paralysis with succinylcholine increased Response entropy and EMG but not State entropy. AB - OBJECTIVE: It is reported that the electromyogram is an indicator of patient arousal during pain stimulation if anesthesia is inadequate. This may not be true during recovery from succinylcholine induced paralysis. We evaluated State entropy of the electroencephalogram (EEG, 0.8-32 Hz) and Response entropy, a combined measure of the electromyogram (EMG) and EEG (0.8-47 Hz), during recovery from paralysis with succinylcholine. METHODS: Twenty patients were randomized to receive either 0.8% (n = 10) or 1.4% isoflurane (n = 10), with 2 mg/kg succinylcholine administered for paralysis in all patients. State entropy and Response entropy were evaluated using a Datex-Ohmeda Entropy module. Frontal EMG was measured separately by an EEG module. State entropy, Response entropy, and EMG were measured in awake patients, during isoflurane anesthesia and paralysis, and after 100% recovery to train of four stimulation. RESULTS: Response entropy and State entropy decreased from awake levels in a dose related manner during 0.8% or 1.4% isoflurane and succinylcholine. Recovery from succinylcholine significantly increased Response entropy and EMG in 5 of 10 patients with 0.8% isoflurane and 8 of 10 with 1.4% isoflurane without a change in State entropy. CONCLUSION: Although RE and EMG increased during recovery from paralysis with succinylcholine, SE, an indicator of EEG, was not stimulated. EMG activity may not be an indicator of patient arousal after succinylcholine treatment. PMID- 16244844 TI - The changes in pulmonary capillary blood flow and anatomical dead space during pulmonary resection under one-lung ventilation. AB - Pulmonary capillary blood flow (PCBF), anatomical dead space (VDaw) and SpO2 were measured and recorded continuously using NICO (Novametrix Medical Systems Inc, USA) in 2 cases during pulmonary resection under one-lung ventilation (OLV). A pulmonary artery catheter was inserted and continuous CO (CCO) was also measured in case 2. In both cases PCBF decreased by half when one lung was blocked. CCO was unchanged before and after one lung blockade in case 2. During OLV, SpO2, which had initially decreased, gradually increased along with a very slow increase in PCBF. The decreased PCBF obtained with NICO represents the pulmonary blood flow of the ventilated (non-blocked) lung. The gradual increase in PCBF with NICO during OLV may express the compensatory effects of hypoxic pulmonary vasoconstriction. By measuring VDaw before, during and after one-lung ventilation, we were able to calculate the bronchial volume of the non-ventilated lung and that of the resected lung. PMID- 16244845 TI - Use of the Patient State Index (PSI) to assist in the diagnosis of perioperative neurological injury and brain death. AB - We present a series of three postoperative cases that were admitted to a cardiothoracic intensive care unit (ICU) after major surgery. Due to the possible presence of residual postoperative neuromuscular blockade after surgery, a processed electroencephalograph (EEG) was applied prior to starting sedation. This was markedly abnormal in all three cases, and not in keeping with the residual anesthesia. The patients were immediately transported for a CT scan. In all three cases there was severe neurological injury incompatible with survival and end of life decisions were made. Although the utility of quantitative EEG technology, like the Bispectral index (BIS) or Patient State Analyzer (PSA), is becoming better defined in the operating room, the role in the ICU is less clear. We propose that the ICU use of the PSA 4000 may have affected our decision weighing the risk versus benefit of transporting a fresh postoperative case to the radiology suite, expedited the neurological diagnosis, and may have reduced overall ICU resource utilization. PMID- 16244847 TI - Optimized brainstem auditory evoked potentials estimation using simulated annealing. AB - In this paper, we use a new approach based on Simulated Annealing for estimating the BAEPs (brainstem auditory evoked potentials). Each BAEP is obtained through a hundred of responses to stimulations. In case of endocochlear pathologies, it has been assumed that these signals could be randomly delayed from a response to another one. In such cases, the application of the averaging method systematically leads to "smoothed" BAEPs, thus complicating both identification and interpretation operations. The method presented in this paper consists in minimizing a non linear criterion in order to obtain an alignment of the various responses, before averaging them. Simulated and experimental results are presented, and compared to those produced by the classical method. PMID- 16244846 TI - A stochastic control program to predict outcome and to support therapeutic decisions: a preliminary report. AB - BACKGROUND AND OBJECTIVES: Early noninvasive hemodynamic monitoring with an outcome predictor and a therapeutic decision support system may be useful to identify and correct hemodynamic deficiencies in emergency patients. The first aim was to apply a stochastic (probability) search and display model to predict outcome as early as possible. The second aim was to explore the usefulness of a therapeutic decision support system to evaluate the relative effectiveness of various therapies. METHODS: A stochastic control and display program based on noninvasive hemodynamic monitoring was applied in 100 consecutive critically ill patients admitted to the emergency department of an inner city public hospital. The program continuously displayed the noninvasive hemodynamic data and the patient's predicted survival probability (SP) that was based on the patient's diagnosis, covariates, and hemodynamic data. The accuracy of the SP at the initial resuscitation on admission to the emergency department (ED) was evaluated by the actual outcome at hospital discharge. The therapeutic decision support program evaluated the relative effectiveness of various therapies on based on their hemodynamic and SP responses and outcome of patients with similar clinical hemodynamic states. RESULTS: The cardiac index, mean arterial pressure, arterial saturation, transcutaneous oxygen and carbon dioxide tensions were appreciably higher in survivors than in nonsurvivors in the initial resuscitation. Heart rate was higher in the nonsurvivors. The calculated Survival Probability (SP) of survivors averaged 81 +/- 1.4% in the first 24-hour observation period. It was 58 +/- 2.2% for nonsurvivors during this period. Misclassifications were 10/100 or 10%. PMID- 16244848 TI - Intraoperative monitoring using somatosensory evoked potentials. A position statement by the American Society of Neurophysiological Monitoring. AB - OBJECTIVE: To provide an educational service to the intraoperative neurophysiologist community by publishing a position statement by the American Society of Neurophysiological Monitoring on the recommended appropriate and correct use of somatosensory evoked potentials as an intraoperative neurophysiological monitoring tool to protect patient well-being during surgery. This position statement presents the somatosensory evoked potential utilization basis, relevant anatomy, patient preparation, important systemic factors, anesthesia considerations, safety and technical considerations, documentation requirements, neurophysiologist credentials and staffing practice patterns, and monitoring applications for protecting brain, spinal nerve root, peripheral nerve, plexus and spinal cord function. In conclusion, a summary of major recommendations regarding the use of somatosensory evoked potentials in intraoperative neurophysiological monitoring is presented. PMID- 16244850 TI - [Teamwork in the operating theatre. Effect on quality of decision-making]. AB - German hospitals face growing economic pressure. Due to the reimbursement system with diagnosis-related groups (DRGs), revenues from capitation fees have to be earned by each clinic. This leads to minimization of resources for every single case. Cost effectiveness is also realised through shorter hospital stays, reduction of hospital beds and a steady rise in cases per year. As a consequence, all employees are confronted with an increasing workload. Compensation of these working conditions can be achieved by human engineering. Despite statements from politicians and hospital leadership in order to cope with this situation, reality is far away: overwork, shorter or missing recreation periods, pressure of time and low income of staff members have turned hospitals into a deterring workplace. If satisfaction of elementary needs such as adherence to break times, is not achieved shortly, politicians and hospital leadership will find out that without motivated and satisfied staff there will not be efficient performance. The present article addresses this issue and gives examples for increasing efficiency through motivation. Focus is on cooperation in the operation room, advantages and risks of teamwork and auxiliary measures for improvement. The aim is to underline how complex and fragile working in an operation room proceeds and how little is done to support this process. Finally, examples are described which improve teamwork, motivation, efficiency and efficacy. PMID- 16244849 TI - [Preoperative psychoeducation for a severely traumatized child]. AB - N., an 8-year-old girl with a long term history of severe sexual abuse and maltreatment with consecutive mental disorders, was scheduled for surgery of clubfeet. The team consisting of an anesthesiologist, a psychologist and a pediatrician decided to prepare the girl for surgery without time pressure. In a first step we applied psychoeducation to obtain a certain degree of confidence. Psychoeducation is a special form of psychological treatment aiming to define cause and goal of surgery together with the patient and relatives. In a further step the anesthesiologist familiarized her with the surgical procedure and the operating theatre. Finally a careful anesthesia with a pain-free postoperative period was applied. Even extremely traumatized children can be prepared for surgery by the help of psychoeducative methods, without time pressure, a careful and pain-free anesthesia and an experienced team of doctors. PMID- 16244851 TI - [The German Telephone Health Survey 2003: an instrument of federal health reporting]. AB - The Robert Koch Institute has carried out health surveys at irregular intervals since the 1980s . Internationally telephone surveys have been used to supplement examination surveys for some time. Health interview surveys by telephone provide a timely and inexpensive possibility to build up a time series on health while being flexible enough to cover new health aspects. Health telephone surveys will also serve as a module of a coming health-monitoring system in Germany, which is currently under discussion. From September 2002 to March 2003, 8,313 German speaking people randomly selected from the resident population aged 18 years and over were asked about topics such as chronic diseases, health behaviour and the extent to which they make use of health care services and others. PMID- 16244852 TI - [Clinical experience with the Siremobil Iso-C(3D) imaging system in pelvic surgery]. AB - In pelvic and acetabular surgery intraoperative control of reduction and implant position is sometimes hard to achieve with conventional C-arm technology. The Siemens C-arm Iso-C(3D) imaging system enables axial cuts and two- or three dimensional reconstructions to be generated. Following the good experience in surgery of the spine and extremities, its clinical applicability in pelvic surgery was evaluated in 30 patients in a prospective clinical trial. In all 20 patients with acetabular fractures reduction quality and implant position could be well assessed. In one postoperative examination an intraarticular screw placement was evident, which intraoperatively was not as clear and was revised. In one procedure an intraarticular fragment was visualized, and was extracted in the same procedure. In one procedure the use of the Iso-C(3D) system enabled the approach to be limited. In all ten pelvic ring injuries the osseous structures in the posterior pelvic ring could be visualized with an adequate image quality. Reduction quality and implant position could be assessed in all open and closed procedures. On two occasions the SI screws were navigated based upon the Iso C(3D) dataset. Overall the use of the Iso-C(3D) system was successful in all cases. Image quality, which is clearly inferior to that of CT, was sufficient for the assessment of reduction quality and implant position. One revision was indicated, and one avoided. However, in spite of its advantages in pelvic surgery, handling, sterile covering and data transfer need to be improved. For bilateral pelvic injuries the image size is too small. PMID- 16244853 TI - Enterococcal prosthetic valve infective endocarditis: report of 45 episodes from the International Collaboration on Endocarditis-merged database. AB - Enterococcal prosthetic valve infective endocarditis (PVE) is an incompletely understood disease. In the present study, patients with enterococcal PVE were compared to patients with enterococcal native valve endocarditis (NVE) and other types of PVE to determine differences in basic clinical characteristics and outcomes using a large multicenter, international database of patients with definite endocarditis. Forty-five of 159 (29%) cases of definite enterococcal endocarditis were PVE. Patients with enterococcal PVE were demographically similar to patients with enterococcal NVE but had more intracardiac abscesses (20% vs. 6%; p=0.009), fewer valve vegetations (51% vs. 79%; p<0.001), and fewer cases of new valvular regurgitation (12% vs. 45%; p=0.01). Patients with either enterococcal PVE or NVE were elderly (median age, 73 vs. 69; p=0.06). Rates of in hospital mortality, surgical intervention, heart failure, peripheral embolization, and stroke were similar in both groups. Patients with enterococcal PVE were also demographically similar to patients with other types of PVE, but mortality may be lower (14% vs. 26%; p=0.08). Notably, 93% of patients with enterococcal PVE came from European centers, as compared with only 79% of patients with enterococcal NVE (p=0.03). Thus, patients with enterococcal PVE have higher rates of myocardial abscess formation and lower rates of new regurgitation compared to patients with enterococcal NVE, but there are no differences between the groups with regard to surgical or mortality rates. In contrast, though patients with enterococcal PVE and patients with other types of PVE share similar characteristics, mortality is higher in the latter group. Importantly, the prevalence of enterococcal PVE was higher in the European centers in this study. PMID- 16244854 TI - Optimal antibiotic treatment in severe pneumococcal pneumonia--time for real answers. PMID- 16244855 TI - Optimization of a protective medium for enhancing the viability of freeze-dried Lactobacillus delbrueckii subsp. bulgaricus based on response surface methodology. AB - Response surface methodology (RSM) was used to optimize a protective medium for enhancing the cell viability of Lactobacillus delbrueckii subsp. bulgaricus LB14 during freeze-drying. Using a previous Plackett-Burman design, it was found that sucrose, glycerol, sorbitol and skim milk were the most effective freeze-drying protective agents for L. bulgaricus LB14. A full factorial central composite design was applied to determine the optimum levels of these four protective agents. The experimental data allowed the development of an empirical model (P<0.0001) describing the inter-relationships between the independent and dependent variables. By solving the regression equation, and analyzing the response surface contour and surface plots, the optimal concentrations of the agents were determined as: sucrose 66.40 g/L, glycerol 101.20 g/L, sorbitol 113.00 g/L, and skim milk 130.00 g/L. L. bulgaricus LB14 freeze-dried in this medium obtained a cell viability of up to 86.53%. PMID- 16244856 TI - Mapping of a spontaneous mutation for early flowering time in maize highlights contrasting allelic series at two-linked QTL on chromosome 8. AB - Only a few mutations affecting flowering time have been detected in maize. We analyzed a spontaneous early mutation, vgt-f7p, which appeared during production of the inbred line F7. This mutation shortens the time from planting to flowering by about 100 growing degree days (GDD), and reduces the number of nodes. It therefore seems to affect the timing of meristem differentiation from a vegetative to a reproductive state. It was mapped to a 6 cM confidence interval on chromosome 8, using a QTL mapping approach. QTL analysis of a mapping population generated by crossing the mutant F7 line (F7p) and the Gaspe flint population showed that vgt-f7p is probably allelic to vgt1, a QTL described in previous studies, and affects earliness more strongly than the Gaspe allele at vgt1. Global analysis of the QTL in the region suggested that there may be two consensus QTL, vgt1 and vgt2. These two QTL have contrasting allelic effects: rare alleles conferring extremely early flowering at vgt1 vs. greater diversity and milder effects at locus vgt2. Finally, detailed syntenic analysis showed that the vgt1 region displays a highly conserved duplicated region on chromosome 6, which also plays an important role in maize flowering time variation. The cloning of vgt1 should, therefore, also facilitate the analysis of the molecular basis of variation due to this second region. PMID- 16244857 TI - Developmental and neurobehavioral effects of perinatal exposure to polychlorinated biphenyls in mice. AB - Because behavioral deficits associated with gestational exposure to polychlorinated biphenyls (PCBs) have been a concern, we studied the developmental and neurobehavioral effects of perinatal exposure to Aroclor 1254 (A1254), a commercial mixture of PCBs, in mice. The PCB mixture (A1254; 0, 6, 18, and 54 mg/kg body weight) was administered to pregnant mice (C57BL/6Cr) every 3 days by gavage from gestational day (GD) 6 to postnatal day (PND) 20. Compared with the control, treatment with A1254 did not alter the maternal body weight during the gestation and lactation periods. The body weight of the offspring did not differ among treatments. To assess the effects on offspring following such exposure, physical and neurobehavioral development (i.e., pinna detachment, hair growth, eye opening, incisor eruption, grasp reflex, righting reflex, walking, negative geotaxis, and cliff avoidance) was observed before weaning. At PND 7, poor adult-like responses in negative geotaxis were observed in all exposed groups. When the offspring were at 8-week old, the PCB-treated (18 mg/kg body weight) mice showed a decreased walking speed in the open-field test, and a prolonged time to reach the platform in the water maze test. Spontaneous locomotion activity was not affected by PCB exposure at 9 weeks . These results showed that perinatal exposure to PCBs produces several behavioral alterations in mice. Although dose-dependent changes were not observed, the neurobehavioral effects such as a decreased walking speed in the open-field test and a prolonged time to reach the platform in the water maze test remained in adulthood after the seeming recovery from the transient delay in development before weaning. PMID- 16244858 TI - Ochratoxin A secretion by ATP-dependent membrane transporters in Caco-2 cells. AB - The ATP-dependent membrane transporters, P-gp, MRP2 and BCRP, localized in the luminal membranes of the intestines, liver and kidney, counteract absorption and increase excretion of xenobiotics and drugs. Previously, it has been suggested that the mycotoxin ochratoxin A (OTA) is a substrate for ATP-dependent transporters, and hence the absorption and secretion of OTA in the Caco-2 cell model was investigated. To this end, Caco-2 cells were cultured as confluent monolayers in bicameral inserts and the transepithelial transport of the mycotoxin was assessed. Caco-2 cells secreted OTA to the luminal side in a concentration-dependent manner. This secretory permeability was higher than the absorptive permeability, while the absorptive permeability remained constant for all OTA concentrations tested. The secretion decreased and absorption increased in the presence of the MRP-inhibitor MK571, the P-gp and BCRP inhibitor GF120918, and the BCRP-inhibitor Ko143, suggesting that the secretion of OTA is mediated by MRP2 and BCRP. Cyclosporine A also decreased the secretory permeability, but did not affect absorptive permeability, while PSC833 did neither change absorption nor secretion of OTA. Hence it can be suggested that OTA is a substrate for MRP2 as well as BCRP. These findings are of interest in evaluating mycotoxin absorption after oral ingestion, tissue distribution and particularly excretion pathways, including renal, biliary and mammary gland excretion. PMID- 16244859 TI - Solid-phase extraction and purification for the quantification of polycyclic aromatic hydrocarbon metabolites in fish bile. AB - An analytical protocol including solid-phase extraction and purification is described for the individual quantification of polycyclic aromatic hydrocarbon metabolites (hydroxylated PAHs) in liquid biological matrices such as plasma and bile. The method consists in an enzymatic deconjugation followed by a solid-phase extraction on a C18 cartridge and by a cleanup on an NH2 cartridge. Extracts are then submitted to a derivatization step before gas chromatography/mass spectrometry (GC/MS) analysis. The quantification of PAH metabolites is ensured by adding an internal standard, 1-hydroxypyrene deuterated, at the beginning of the protocol. Recoveries obtained for the entire protocol were for the major part of the compounds between 96 and 70%. However, recoveries were not so satisfying concerning 2-hydroxybiphenyl and especially 3-hydroxybenzo(a)pyrene, with 62 and 36% respectively. Finally, the protocol was applied to different fish bile samples and showed its good applicability to fish bile samples. The NH2 cleanup step has been proved to be a very selective purification step, necessary to remove most of the bile pigments before GC/MS injection. Different PAH metabolites could be detected in those natural samples and their quantification allowed us to distinguish different levels of fish exposure. PMID- 16244860 TI - Genetic organization of a plasmid from an industrial wastewater bioreactor. AB - Pseudomonas strain CT14 was isolated from activated sludge. Strain CT14 contained a 55, 216 bp plasmid that was characterized by sequence analysis. The plasmid had a modular structure with 51 open reading frames (ORFs) that were distributed between two clearly demarcated domains. Domain I primarily contained genes for plasmid-related functions and a novel origin of replication. Domain II bore evidence of extensive transposition and recombination. Domain II contained several genes from a meta-cleavage pathway for aromatic rings. These genes appeared to have been recruited from different hosts. This observation suggests that sequencing pCT14 may have revealed an intermediate stage in the evolution of a new assemblage of meta-cleavage pathway genes. PMID- 16244861 TI - An unusual late presentation of a congenital diaphragmatic hernia. AB - An unusual and late-presenting case of left Bochdalek diaphragmatic hernia is presented to highlight the fact that the timing and mechanism of presentation of the hernia remains obscure. In this case there were two previous admissions, initially for a left upper lobe lobectomy for congenital lobar emphysema and a subsequent admission for bronchiolitis. He then presented at the age of nine months, with acute respiratory distress and was diagnosed to have a left diaphragmatic hernia. This is the first case of a late- presenting congenital diaphragmatic hernia reported in association with a previous ipsilateral lobectomy for congenital lobar emphysema. A brief review of late -presenting congenital diaphragmatic hernias is given and an alternative theory is proposed for the mechanism of late presentation. PMID- 16244862 TI - Late-onset hemothorax after the Nuss procedure for funnel chest. AB - A 4-year-and-3-month-old boy with funnel chest underwent the Nuss procedure. He had an uneventful intraoperative and postoperative course, and was discharged on the tenth day of hospitalization. He developed chest pain while playing on the 29th day after surgery, and was diagnosed with right hemothorax. He was followed conservatively and the hemothorax disappeared. PMID- 16244863 TI - Protective effects of trapidil in lung after abdominal aorta induced ischemia reperfusion injury: an experimental study. AB - We aimed to investigate the protective effects of trapidil after the occlusion of abdominal aorta and the reperfusion injury in lung. Eighteen New Zealand albino rabbits were used in the study. In six animals [group 1, ischemia-reperfusion (IR) group], the abdominal aorta was exposed and a microvascular clamp was placed in the infrarenal abdominal aorta for 60 min. After the ischemic period, the microvascular clamp was removed and reperfusion was provided for 2 h. After the reperfusion period, the lungs were removed carefully and specimens were prepared for histopathological and biochemical studies in appropriate conditions. In group 2 (study group), trapidil (Rocarnal, Rentschler-UCB GmbH, Kerpen, Germany) was administered intraperitoneally as a single dose 1 h prior to trial, the IR procedure was performed and lung specimens were prepared similar to group 1. In group 3 (sham group), the infrarenal abdominal aorta was exposed and lung specimens were prepared for histopathological and biochemical studies at the end of the study. Histopathological changes, malondialdehyde (MDA), nitric oxide (NO) and total sulfhydryl group (T-SH) levels were evaluated. There was a statistical difference between the IR group and study group regarding NO and MDA levels (P < 0.05 and P < 0.01, respectively), but this was not detected between the IR group and the sham group (P > 0.05). There was no statistical difference among the three groups regarding T-SH levels (P > 0.05). While a statistical difference was found between the sham group and study group in the NO level (P < 0.05), no statistical difference was found in the MDA level (P > 0.05). There was a statistical difference in interstitial edema, PMN infiltration and hemorrhage scores among the groups (P < 0.05). There was a statistical difference between the IR group and study group in PMN infiltration (P < 0.05), but this was not detected between the groups in interstitial edema and hemorrhage scores (P > 0.05). There was a statistical difference between IR group and sham group in interstitial edema, PMN infiltration and hemorrhage scores (P < 0.05). Statistical difference was found between the sham group and study group in interstitial edema and hemorrhage scores (P < 0.05), but not in PMN infiltration (P > 0.05). CONCLUSIONS: Infrarenal abdominal aortic occlusion and reperfusion causes lung injury. We conclude that trapidil has preventive effects in the lung tissue after IR injury. PMID- 16244864 TI - Functional changes after physiotherapy in fecal incontinence. AB - BACKGROUND: Physiotherapy is a common treatment option in patients with fecal incontinence. Although physiotherapy may result in relief of symptoms, to what extent improvement is associated with changes in anorectal function is still unclear. AIM: The aim of the present study was to investigate prospectively how anorectal function changes with physiotherapy and whether these changes are related to changes in fecal incontinence score. METHODS: Consenting consecutive patients (n=266) with fecal incontinence (91% women; mean age, 59 years) underwent anorectal manometry, anal and rectal mucosal sensitivity measurements, and rectal capacity measurement at baseline and after nine sessions of standardized pelvic floor physiotherapy. These findings were compared with changes in Vaizey incontinence score. RESULTS: On follow-up 3 months after physiotherapy, squeeze pressure (p=0.028), as well as urge sensation threshold (p=0.046) and maximum tolerable volume (p=0.018), had increased significantly. The extent of improvement was not related to age, duration of fecal incontinence, menopause, and endosonography findings. All other anorectal functions did not change. An improvement in the Vaizey score was moderately correlated with an increase in incremental squeeze pressure (r=0.14, p=0.04) and a decrease in anal mucosal sensitivity threshold (r=0.20, p=0.01). CONCLUSIONS: Physiotherapy improves squeeze pressure, urge sensation, and maximum tolerable volume. However, improved anorectal function does not always result in a decrease in fecal incontinence complaints. PMID- 16244865 TI - Thallus morphology and optical characteristics affect growth and DNA damage by UV radiation in juvenile Arctic Laminaria sporophytes. AB - Growth of young sporophytes of the brown algae Laminaria digitata, L. saccharina and L. solidungula from Spitsbergen were measured in the laboratory after being exposed for 21 days to either photosynthetically active radiation (PAR = P) or to full light spectrum (PAR + UV-A + UV-B = PAB) using of cutoff glass filters. The plants were grown at 8+/-2 degrees C and 16 h light : 8 h dark cycles with 6 h additional ultraviolet radiation (UVR) exposure in the middle of the light period. Growth was measured every 10 min using growth chambers with online video measuring technique. Tissue morphology and absorption spectra were measured in untreated young sporophytes while chlorophyll (Chl) a content and DNA damage were measured in treated thalli at the end of the experiment. In all species, growth rates were significantly higher in sporophytes exposed to P alone compared to sporophytes exposed to PAB. Tissue DNA damage is dependent on thallus thickness and absorption spectra characteristics of pigments and UV-absorbing compounds. In sporophytes exposed to UVR, energy demands for repair of DNA damage and synthesis of UV-absorbing compounds for protection effectively diverts photosynthate at the expense of growth. Photosynthetic pigment was not significantly different between treatments suggesting a capacity for acclimation to moderate UVR fluence. The general growth pattern in sporophytes exposed to P alone showed an increasing growth rate from the onset of light (0500-0900 hours) to a peak at the middle of the light phase (0900-1500 hours), a decline towards the end of the light phase (1500-2100 hours) and a minimum "low" growth in the dark (2100-0500 hours) relative to growth during the entire light phase. Under PAB, different growth patterns were observed such as growth compensation at night in L. digitata, delayed growth recovery in L. saccharina and minimal but continuous growth in L. solidungula. Growth as an integrative parameter of all physiological processes showed that the effect of UVR is correlated to the depth distribution of these species. PMID- 16244866 TI - UFO in the Arabidopsis inflorescence apex is required for floral-meristem identity and bract suppression. AB - The UNUSUAL FLORAL ORGANS (UFO) gene of Arabidopsis encodes an F-box protein required for the determination of floral-organ and floral-meristem identity. Mutation of UFO leads to dramatic changes in floral-organ type which are well characterized whereas inflorescence defects are more subtle and less understood. These defects include an increase in the number of secondary inflorescences, nodes that alternate between forming flowers and secondary inflorescences, and nodes in which a single flower is subtended by a bract. Here, we show how inflorescence defects correlate with the abnormal development of floral primordia and establish a temporal requirement for UFO in this process. At the inflorescence apex of ufo mutants, newly formed primordia are initially bract like. Expression of the floral-meristem identity genes LFY and AP1 are confined to a relatively small adaxial region of these primordia with expression of the bract-identity marker FIL observed in cells that comprise the balance of the primordia. Proliferation of cells in the adaxial region of these early primordia is delayed by several nodes such that primordia appear "chimeric" at several nodes, having visible floral and bract components. However, by late stage 2 of floral development, growth of the bract generally ceases and is overtaken by development of the floral primordium. This abnormal pattern of floral meristem development is not rescued by expression of UFO from the AP1 promoter, indicating that UFO is required prior to AP1 activation for normal development of floral primordia. We propose that UFO and LFY are jointly required in the inflorescence meristem to both promote floral meristem development and inhibit, in a non-cell autonomous manner, growth of the bract. PMID- 16244867 TI - Early responses to Nod factors and mycorrhizal colonization in a non-nodulating Phaseolus vulgaris mutant. AB - Legumes can acquire nitrogen through a symbiotic interaction with rhizobial bacteria. The initiation of this process is determined by a molecular dialogue between the two partners. Legume roots exude flavonoids that induce the expression of the bacterial nodulation genes, which encode proteins involved in the synthesis and secretion of signals called Nod factors (NFs). NFs signal back to the plant root and trigger several responses, leading to bacterial invasion and nodule formation. Here, we describe the molecular and cellular characterization of a Phaseolus vulgaris non-nodulating mutant (NN-mutant). Root hair cells of the NN-mutant plant respond with swelling and branching when inoculated with Rhizobium etli, albeit without curling induction. Furthermore, neither initiation of cell division in the outer cortex, nor entrapment of bacteria nor infection thread formation was observed. Both the bean wild-type and the NN-mutant responded with elevated intracellular calcium changes in the root hairs. Although the NN-mutant is deficient in early nodulin gene expression when inoculated with R. etli, it can be effectively colonized by arbuscular mycorrhizal fungi (Glomus intraradices). Our data indicate that the P. vulgaris NN-mutant is not blocked at the NFs early perception stage, but at later downstream stages between Ca(2+) signaling and early nodulin induction. This supports the idea that both microsymbionts are perceived and trigger different downstream pathways in the host plant. PMID- 16244869 TI - Biphasic solitary fibrous tumour: a report of two cases with epithelioid features. AB - We present two cases of solitary fibrous tumour (SFT) showing biphasic morphology with a spectrum of malignant epithelioid components. Slides prepared from formalin-fixed and paraffin-embedded tissue from both cases were stained with haematoxylin and eosin and by immunohistochemistry. Interphase fluorescent in situ hybridisation studies were performed in both cases using paraffin-embedded tissue to look for the t(X;18) translocation, thereby to exclude synovial sarcoma. Both cases showed biphasic morphology with some areas having typical benign spindled SFT morphology (including CD34 expression) and other areas having a malignant epithelioid appearance. In one of the cases, the epithelioid area, which was well circumscribed and showed packeting of cell groups, demonstrated expression of cytokeratin and epithelial cadherin but not of CD34. In the second case, the immunophenotype of the epithelioid component was similar to that of the benign SFT component. These findings suggest that epithelioid change in SFT shows a range of differentiation at one end, similar to that of a standard SFT, and at the other end, possibly acquiring epithelial characteristics. PMID- 16244868 TI - The Arabidopsis LHP1 protein is a component of euchromatin. AB - The HP1 family proteins are involved in several aspects of chromatin function and regulation in Drosophila, mammals and the fission yeast. Here we investigate the localization of LHP1, the unique Arabidopsis thaliana HP1 homolog known at present time, to approach its function. A functional LHP1-GFP fusion protein, able to restore the wild-type phenotype in the lhp1 mutant, was used to analyze the subnuclear distribution of LHP1 in both A. thaliana and Nicotiana tabacum. In A. thaliana interphase nuclei, LHP1 was predominantly located outside the heterochromatic chromocenters. No major aberrations were observed in heterochromatin content or chromocenter organization in lhp1 plants. These data indicate that LHP1 is mainly involved in euchromatin organization in A. thaliana. In tobacco BY-2 cells, the LHP1 distribution, although in foci, slightly differed suggesting that LHP1 localization is determined by the underlying genome organization of plant species. Truncated LHP1 proteins expressed in vivo allowed us to determine the function of the different segments in the localization. The in foci distribution is dependent on the presence of the two chromo domains, whereas the hinge region has some nucleolus-targeting properties. Furthermore, like the animal HP1beta and HP1gamma subtypes, LHP1 dissociates from chromosomes during mitosis. In transgenic plants expressing the LHP1-GFP fusion protein, two major localization patterns were observed according to cell types suggesting that localization evolves with age or differentiation states. Our results show conversed characteristics of the A. thaliana HP1 homolog with the mammal HP1gamma isoform, besides specific plant properties. PMID- 16244871 TI - Relationship between mRNA levels and protein accumulation in a chloroplast promoter-mutant of Chlamydomonas reinhardtii. AB - The photosynthetic chloroplast mutant G64 of Chlamydomonas reinhardtii was shown to contain a single point mutation within the 5' region of the psbD gene encoding the D2 protein of the photosystem II reaction center. The mutation affects the sequence element TATAATAT which has previously been hypothesized to function as the psbD promoter. Run-on analysis confirmed that transcription of psbD in the mutant was reduced to approximately 10% of the wild-type level. However, psbD mRNA accumulated to approximately 35%, despite the prominent decrease in RNA synthesis. This suggests that RNA-stabilization effects can compensate to some extent for a reduction in transcriptional activity. Interestingly, a direct correlation between transcript levels and the accumulation of the psbD gene product, the D2-protein, was observed in G64. The data suggest that posttranscriptionally acting regulatory factors determine the rate-limiting steps of chloroplast psbD gene expression. PMID- 16244870 TI - Xq25 and Xq26 identify the common minimal deletion region in malignant gastroenteropancreatic endocrine carcinomas. AB - Loss of heterozygosity (LOH) for markers on X chromosome are associated with malignancy in endocrine tumors of the stomach and pancreas. The aim of this work is to investigate low-grade, well-differentiated endocrine carcinomas (WDEC) vs high-grade, poorly differentiated endocrine carcinomas (PDEC) of the gastroenteropancreatic (GEP) tract for common deletion regions on X chromosome. We performed a comparative allelotyping analysis with 24 highly polymorphic markers for the X chromosome in 12 WDECs and 5 PDECs. Overall, the LOH frequency in all informative loci investigated was 59% in primary and 61% in metastasis, with a significantly higher rate in PDECs than in WDECs (p<0.015 for primary and p<0.00005 for metastasis). In both WDECs and PDECs, the small Xq25 region as defined by DXS8059, DXS8098, and DXS8009 markers showed higher LOH rate as compared to the rest of the chromosome markers (p<0.04). In addition, LOH was very frequently elevated also in DXS294 and in DXS102 loci mapping the chromosomal region Xq26. In no instances differences were found between primary tumors and metastases. Methylation analysis revealed that Xq25 loss preferentially occurred on the inactive X chromosome, a feature in agreement with findings from other human cancers suggesting escape of tumor suppressor genes to X chromosome inactivation at this region. Overall, our data indicate that the two chromosomal regions, Xq25 and Xq26, may participate to the malignant progression of GEP endocrine carcinomas. PMID- 16244872 TI - Genome-wide SNP discovery and linkage analysis in barley based on genes responsive to abiotic stress. AB - More than 2,000 genome-wide barley single nucleotide polymorphisms (SNPs) were developed by resequencing unigene fragments from eight diverse accessions. The average genome-wide SNP frequency observed in 877 unigenes was 1 SNP per 200 bp. However, SNP frequency was highly variable with the least number of SNP and SNP haplotypes observed within European cultivated germplasm reflecting effects of breeding history on genetic diversity. More than 300 SNP loci were mapped genetically in three experimental mapping populations which allowed the construction of an integrated SNP map incorporating a large number of RFLP, AFLP and SSR markers (1,237 loci in total). The genes used for SNP discovery were selected based on their transcriptional response to a variety of abiotic stresses. A set of known barley abiotic stress QTL was positioned on the linkage map, while the available sequence and gene expression information facilitated the identification of genes potentially associated with these traits. Comparison of the sequenced SNP loci to the rice genome sequence identified several regions of highly conserved gene order providing a framework for marker saturation in barley genomic regions of interest. The integration of genome-wide SNP and expression data with available genetic and phenotypic information will facilitate the identification of gene function in barley and other non-model organisms. PMID- 16244874 TI - Mapping autosomal dominant progressive limb-girdle myopathy with bone fragility to chromosome 9p21-p22: a novel locus for a musculoskeletal syndrome. AB - Progressive myopathy of a limb-girdle distribution and bone fragility is a rare autosomal dominant disorder of unknown etiology. Affected individuals, within this family, present with various combinations of progressive muscle weakness, easy fracturing, and poor healing of long bones. Additional features include premature graying with thin hair, thin skin, hernias, and clotting disorders. Electromyograms show myopathic changes and biopsies reveal non-specific myopathic changes. Skeletal radiographs demonstrate coarse trabeculation, patchy sclerosis, cortical thickening, and narrowing of medullary cavities. We report genetic mapping of this disorder to chromosome 9p21-p22 in a multigenerational family. A genome-wide scan for the disease locus obtained a maximal LOD score of 3.74 for marker GATA87E02 N (D9S1121). Haplotype analysis localized the disease gene within a 15 Mb interval flanked by markers AGAT142P and GATA5E06P. This region also localizes diaphyseal medullary stenosis with malignant fibrous histiocytoma (DMS-MFH). Identification of the disease gene will be necessary to understand the pathogenesis of this complex disorder. PMID- 16244873 TI - The neurobeachin gene spans the common fragile site FRA13A. AB - Common fragile sites are normal constituents of chromosomal structure prone to chromosomal breakage. In humans, the cytogenetic locations of more than 80 common fragile sites are known. The DNA at 11 of them has been defined and characterized at the molecular level. According to the Genome Database, the common fragile site FRA13A maps to chromosome band 13q13.2. Here, we identify the precise genomic position of FRA13A, and characterize the genetic complexity of the fragile DNA sequence. We show that FRA13A breaks are limited to a 650 kb region within the neurobeachin (NBEA) gene, which genomically spans approximately 730 kb. NBEA encodes a neuron-specific multidomain protein implicated in membrane trafficking that is predominantly expressed in the brain and during development. PMID- 16244875 TI - Impaired transcriptional upregulation of Parkin promoter variant under oxidative stress and proteasomal inhibition: clinical association. AB - We provided data to show that the transcriptional activity of wildtype -258T in the parkin promoter region was significantly higher than the -258G variant in human cell lines. The transcriptional activity of wildtype -258T was significantly increased under oxidative stress by hydrogen peroxide, but this was not observed for the -258G variant. The transcriptional upregulation was significantly higher for wildtype -258T compared to -258G variant at 0.1, 0.2 and 0.4 mM of hydrogen peroxide. Similar results were obtained when the cells were treated with a proteasome inhibitor, MG132.Furthermore, in a case control study involving 753 subjects, we demonstrated that the parkin promoter -258G variant was associated with an increased risk of sporadic Parkinson's disease (PD) in the elderly ethnic Chinese population. Our clinical and laboratory data provide corroborative evidence that some older individuals who have the -258G variant may have a higher risk of developing PD. PMID- 16244877 TI - The attributable mortality of acute renal failure in critically ill patients with liver cirrhosis. AB - OBJECTIVE: To determine outcome and mortality risk related to acute renal failure (ARF) in critically ill patients with cirrhosis. DESIGN AND SETTING: A retrospective cohort analysis and two independent case-control analyses in a medical ICU. PATIENTS: 41 and 32 patients who developed mild and severe ARF, respectively, matched (1:2 ratio) with cirrhotic patients without ARF during their ICU stay. MEASUREMENTS AND RESULTS: Cirrhotic patients with ARF had higher MELD, APACHE II, and SOFA scores at baseline that those without ARF. They had more respiratory failure and cardiovascular failure during ICU stay, longer stay in ICU, and a greater crude hospital mortality rate (65% vs. 32%). Multivariate survival analysis identified ARF (hazard ratio, HR, 4.1), alcohol abuse or dependency, and severe sepsis or septic shock as independent predictors of death. In case-control studies both mild and severe ARF were independently associated with mortality (HR, 2.6, and 4.2, respectively). Cirrhotic patients with mild ARF patients had a higher risk of death than those without ARF (relative risk, RR, 2.0). Severe ARF was associated with an increase matched risk of death (RR 2.6), higher mortality of 51%, and higher risk-adjusted mortality rate (2.1 vs. 0.9). CONCLUSIONS: ICU patients with liver cirrhosis still have a high crude mortality. In this specific population ARF is associated with an excess mortality, depending on the severity of renal dysfunction. PMID- 16244878 TI - The use of severity scores in the intensive care unit. PMID- 16244876 TI - Proprotein convertases furin and PC5: targeting atherosclerosis and restenosis at multiple levels. AB - Several growth factors, chemokines, adhesion molecules, and proteolytic enzymes important for cell-cell/cell-matrix interactions in atherosclerosis and restenosis are initially synthesized as inactive precursor proteins. Activation of proproteins to biologically active molecules is regulated by limited endoproteolytic cleavage at dibasic amino acid residues. This type of activation typically requires the presence of suitable proprotein convertases (PCs). The PC isozymes furin and PC5 are expressed in human atherosclerotic lesions and have been found to be up-regulated, following vascular injury in animal models in vivo. In vitro, these PCs can regulate vascular smooth muscle cell and macrophage functions and signaling events, through activation of pro-alpha-integrins and/or pro-membrane-type matrix metalloproteinases. Integrins link the cytoskeleton with the extracellular matrix and mediate bidirectional signaling and mechanotransduction, whereas matrix metalloproteinases are the major matrix degrading enzymes. Both activities are required for cell recruitment to the intima. Furthermore, cleavage of extracellular matrix molecules by matrix metalloproteinases potentially contributes to weakening of the fibrous cap, promoting plaque rupture. Based on these recent in vitro and in vivo data, furin and PC5 are potential contributors to the initiation, progression, and complications of atherosclerosis and restenosis. Targeting these PCs may provide future anti-atherosclerotic therapies. PMID- 16244879 TI - Accuracy of clinical presentation for differentiating bacterial from viral meningitis in adults: a multivariate approach. AB - OBJECTIVE: To determine whether bacterial (BM) and viral (VM) meningitis can be differentiated based on initial clinical presentation. DESIGN AND SETTING: Retrospective cohort study in a medical emergency department and intensive care unit in a university hospital. PATIENTS: 144 adults, including 90 with confirmed BM and 54 unpretreated VM. MEASUREMENTS AND RESULTS: Symptoms, examination findings, paraclinical data, and clinical outcome were assessed. Severity was defined by the presence at referral of one of the following criteria: altered consciousness, seizures, focal neurological findings, and shock. After univariate analyses we performed stepwise logistic regression to determine predictors for BM available at referral (except for CSF Gram stain) and logistic regression using previously validated CSF cutoffs. Univariate methods identified the presence of one sign of severity as the most important predictor for BM (sensitivity 0.989, specificity 0.981, positive predictive value 0.989, negative predictive value 0.981, odds ratio 4,770) and showed that CSF results differ in BM and in VM (except for CSF glucose). Logistic regression analysis revealed severity and CSF absolute neutrophil count as the two predictors of BM (R2=0.876). Logistic analysis showed that BM was related to severity (beta=6.46+/-1.27) and a CSF absolute neutrophil count above 1,000/mm3 whereas CSF glucose below 2 mmol/l and CSF protein higher than 2 g/l were not predictive. CONCLUSIONS: The presence of at least one sign of severity at referral and a CSF absolute neutrophil count above 1,000/mm3 mm are predictive of BM. PMID- 16244880 TI - How to differentiate bacterial from viral meningitis. PMID- 16244881 TI - Passive stretching produces Akt- and MAPK-dependent augmentations of GLUT4 translocation and glucose uptake in skeletal muscles of mice. AB - Muscle contraction is accompanied by passive stretching or deformation of cells and tissues. The present study aims to clarify whether or not acute passive stretching evokes glucose transporter 4 (GLUT4) translocation and glucose uptake in skeletal muscles of mice. Passive stretching mainly induced GLUT4 translocation from an intracellular membrane-rich fraction (PF5) to a plasma membrane-rich fraction (F2) and accelerated glucose uptake in hindlimb muscles; whereas electrical stimulation, which mimics physical exercise in vivo, and insulin, each induced GLUT4 translocation from an intracellular membrane-rich fraction (PF5) to a fraction rich in plasma membrane (F2), and to one rich in transverse tubules (PF3), along with subsequent glucose uptake. Mechanical stretching increased phosphorylation of Akt and p38 mitogen-activated protein kinase (p38 MAPK), but it had no apparent effect on the activity of AMP-activated protein kinase (AMPK). Electrical stimulation augmented the activity of not only AMPK but also phosphorylation of Akt and p38 MAPK. Our results suggest that passive stretching produces translocation of GLUT4 mainly from the fraction rich in intracellular membrane to that rich in plasma membrane, and that the glucose uptake could be Akt- and p38 MAPK-dependent, but AMPK-independent manners. PMID- 16244882 TI - Study on fish embryo responses to the treatment of cryoprotective chemicals using impedance spectroscopy. AB - Investigations using electrical impedance spectroscopy to measure the responses of fish embryos to the cryoprotective chemicals, methanol and dimethyl sulphoxide (DMSO), were carried out. Zebrafish (Danio rerio) embryos were used as a model to study the newly proposed technique. The normalised permittivity and conductivity changes of the embryos were measured continuously over a 20-min period in a customised embryo-holding chamber. The normalised permittivity and conductivity spectra were obtained during embryo exposure to different concentrations of methanol (1.0, 2.0 and 3.0 M) and DMSO (0.5, 1.0 and 2.0 M) solutions. The results showed significant permittivity and conductivity changes after embryo exposure to methanol and DMSO at the optimum embryo loading level (six embryos). Embryos in different concentrations of methanol and DMSO also resulted in quantitative responses shown in the normalised permittivity and conductivity spectra. The results demonstrated that fish embryo membrane permeability to cryoprotective chemicals could be monitored in real-time. The measurement of permittivity at a lower frequency range (10-10(3) Hz) and conductivity at a higher frequency range (10(4)-10(6) Hz) during fish embryo exposure to cryoprotective chemicals using impedance spectroscopy can be used as a new tool for the fast screening of most effective cryoprotective chemicals. The results from the present study also demonstrated the possibility of quantifying the level of cryoprotective chemicals penetrating the fish embryos. PMID- 16244883 TI - Identification and phylogenetic analysis of 15 MHC class II DRB1 beta1 expressed alleles in a ewe-lamb flock. AB - Ovar-DRB1 is part of the major histocompatibility complex (MHC) class II of sheep and functions by presenting extracellular-derived peptides to the immune system. Although there are over 100 different Ovar-DRB1 DNA sequences reported in GenBank, only two Ovar-DRB1 mRNA sequences have been reported. As a first step in understanding MHC Class II function as it relates to disease progression in sheep, Ovar-DRB1 transcripts encoding the peptide-binding site or the first domain (beta1) of Ovar-DRB1 in a 32-ewe-lamb flock were identified and characterized by using reverse transcriptase-polymerase chain reaction, cloning, sequencing, and phylogenetic analysis. Fourteen new Ovar-DRB1 beta1 cDNA sequences out of a total of 15 Ovar-DRB1 beta1 cDNA sequences in a ewe-lamb flock of 32 sheep were identified. One Ovar-DRB1 beta1 cDNA sequence was 100% identical to M93432, one of the two Ovar-DRB1 mRNA sequences reported in GenBank. Twelve out of 15 Ovar-DRB1 beta1 cDNA sequences were 100% identical to the corresponding previously reported Ovar-DRB1 genomic DNA sequences, indicating that these Ovar DRB1 genomic DNA sequences are also transcribed. One of three of the remaining Ovar-DRB1 beta1 cDNA sequences, DRB1*07012, had a synonymous substitution resulting in an identical deduced amino acid sequence to DRB1*0701. Two of the remaining three Ovar-DRB1 beta1 cDNA sequences had nucleotide differences and subsequent deduced amino acid sequence differences when compared to known Ovar DRB1 beta1 genomic DNA sequences, and therefore, DRB1*0206 and DRB1*0353 represent new Ovar-DRB1 beta1 expressed alleles. Phylogenetic analysis of the 15 Ovar-DRB1 beta1 cDNA sequences revealed that DRB1*0206 had a strong phylogenetic relationship to DRB1*0203, and DRB1*0353 had a strong phylogenetic relationship to DRB1*0303. PMID- 16244884 TI - Genetic transformation of cowpea (Vigna unguiculata L.) and stable transmission of the transgenes to progeny. AB - Cowpeas are nutritious grains that provide the main source of protein, highly digestible energy and vitamins to some of the world's poorest people. The demand for cowpeas is high but yields remain critically low, largely because of insect pests. Cowpea germplasm contains little or no resistance to major insect pests and a gene technology approach to adding insect protection traits is now a high priority. We have adapted features of several legume and other transformation systems and reproducibly obtained transgenic cowpeas that obey Mendelian rules in transmitting the transgene to their progeny. Critical parameters in this transformation system include the choice of cotyledonary nodes from developing or mature seeds as explants and a tissue culture medium devoid of auxins in the early stages, but including the cytokinin BAP at low levels during shoot initiation and elongation. Addition of thiol-compounds during infection and co culture with Agrobacterium and the choice of the bar gene for selection with phosphinothricin were also important. Transgenic cowpeas that transmit the transgenes to their progeny can be recovered at a rate of one fertile plant per thousand explants. These results pave the way for the introduction of new traits into cowpea and the first genes to be trialled will include those with potential to protect against insect pests. PMID- 16244885 TI - Fly pupae and puparia as potential contaminants of forensic entomology samples from sites of body discovery. AB - Fly pupae and puparia may contaminate forensic entomology samples at death scenes if they have originated not from human remains but from animal carcasses or other decomposing organic material. These contaminants may erroneously lengthen post mortem interval estimates if no pupae or puparia are genuinely associated with the body. Three forensic entomology case studies are presented, in which contamination either occurred or was suspected. In the first case, blow fly puparia collected near the body were detected as contaminants because the species was inactive both when the body was found and when the deceased was last sighted reliably. The second case illustrates that contamination may be suspected at particularly squalid death scenes because of the likely presence of carcasses or organic material. The third case involves the presence at the body discovery site of numerous potentially contaminating animal carcasses. Soil samples were taken along transects to show that pupae and puparia were clustered around their probable sources. PMID- 16244886 TI - Expression of hunchback during trunk segmentation in the branchiopod crustacean Artemia franciscana. AB - Comparative studies have shown that some aspects of segmentation are widely conserved among arthropods. Yet, it is still unclear whether the molecular prepatterns that are required for segmentation in Drosophila are likely to be similarly conserved in other arthropod groups. Homologues of the Drosophila gap genes, like hunchback, show regionally restricted expression patterns during the early phases of segmentation in diverse insects, but their expression patterns in other arthropod groups are not yet known. Here, we report the cloning of a hunchback orthologue from the crustacean Artemia franciscana and its expression during the formation of trunk segments. Artemia hunchback is expressed in a series of segmental stripes that correspond to individual thoracic/trunk, genital, and postgenital segments. However, this expression is not associated with the segmenting ectoderm but is restricted to mesodermal cells that associate with the ectoderm in a regular metameric pattern. All cells in the early segmental mesoderm appear to express hunchback. Later, mesodermal expression fades, and a complex expression pattern appears in the central nervous system (CNS), which is comparable to hunchback expression in the CNS of insects. No regionally restricted expression, reminiscent of gap gene expression, is observed during trunk segmentation. These patterns suggest that the expression patterns of hunchback in the mesoderm and in the CNS are likely to be ancient and conserved among crustaceans and insects. In contrast, we find no evidence for a conserved role of hunchback in axial patterning in the trunk ectoderm. PMID- 16244887 TI - PCR-based identification and delineation of members within the Pseudorhabdosynochus lantauensis complex (Monogenea: Diplectanidae). AB - Molecular methods using genetic markers in the nuclear ribosomal DNA (rDNA) were established to identify and distinguish between two members within the Pseudorhabdosynochus lantauensis complex and two morphologically distinct congeners, Pseudorhabdosynochus epinepheli and Pseudorhabdosynochus coioidesis, from different marine fish species and various geographical origins. Supported by selective DNA sequencing, it was demonstrated that the polymerase chain reaction (PCR)-coupled single-strand conformation polymorphism analysis of the first internal transcribed spacer and restriction fragment length polymorphism analysis of a variable region (representing the D1-D3 domains) in the large subunit of rDNA achieved the identification and delineation of all four taxa examined. These PCR-based approaches provide useful complementary tools to traditional methods for the accurate identification of species within the genus Pseudorhabdosynochus (irrespective of developmental stage) and have major implications for studying the ecology, transmission, and population genetic structures of these and other related parasites and for the prevention and control of the diseases they cause. PMID- 16244888 TI - Distribution of adherens junction mediated by VE-cadherin complex in rat spleen sinus endothelial cells. AB - The splenic sinus endothelium regulates the passage of blood cells through the splenic cord. The goal of the present study was to assess the localization of vascular endothelial (VE)-cadherin, beta-catenin, and p120-catenin in the sinus endothelial cells of rat spleen and to characterize the presence and distribution of adherens junction formation mediated by the cadherin-catenin complex. Immunofluorescent microscopy of tissue cryosections demonstrated that VE cadherin, beta-catenin, and p120-catenin were localized in the junctional regions of adjacent endothelial cells. Double-staining immunofluorescent microscopy for VE-cadherin and beta-catenin revealed colocalization at junctional regions. Transmission electron microscopy of thin sections of sinus endothelial cells treated with Triton X-100 clearly showed adherens junctions within the plasma membrane. Adherens junctions were located at various levels in the lateral membranes of adjacent endothelial cells regardless of the presence or absence of underlying ring fibers. Immunogold electron microscopy revealed VE-cadherin, beta catenin, and p120-catenin in the juxtaposed junctional membranes of adjacent sinus endothelial cells. Double-staining immunogold microscopy for VE-cadherin and beta-catenin and for VE-cadherin and p120-catenin demonstrated colocalization to the junctional membranes of adjacent endothelial cells. Immunolabeling was evident at various levels in the lateral junctional membranes and was intermittently observed in the sinus endothelium. These data suggest that adherens junctions, whose formation appears to be mediated by VE-cadherin-catenin complexes, probably regulate the passage of blood cells through the spleen. PMID- 16244889 TI - [How does recruit successfully recruit staff? An investigation of recruitment in the medical service]. AB - In Germany, an increasing shortage of medical doctors has developed in recent years. Today, many clinics must compete to attract qualified MDs. In this study, 60 medical students, 60 doctors working in psychiatry, and 60 doctors working in somatic fields of clinical medicine were interviewed in order to analyse the importance that potential applicants place on the information and offers contained in advertisements seeking medical doctors. In this regard, good working atmosphere in the ward, excellent opportunities for further education, and assistance from nonmedical personnel in documentation and administration got the highest ratings. There were significant differences between psychiatrists and other doctors rating the items "own office" and "permission to take additional nonhospital jobs" such as providing medical opinions. The study shows how advertisements can be improved. PMID- 16244890 TI - [Forensic psychiatry. Its relations to clinical psychiatry and criminology]. AB - A basic task of psychiatry is to identify and treat mentally disordered persons at risk of committing crimes. Psychiatry has an important function in preserving social peace, law, and order. How the psychiatric world handles this duty has changed with time. There have been very important changes from asylums to mental hospitals and from voluntary or involuntary inpatient treatment to outpatient care; but clinical psychiatry cannot give up forensic psychiatry. As a result of developments, inpatient care in mental hospitals often concentrates on crisis management, risk assessment, and risk management. On the other hand, forensic psychiatry has made great efforts in recent decades with special therapies for mentally disturbed criminals and collaborated closely with criminologists in developing instruments for risk assessment and prognosis of repeat offenses. PMID- 16244891 TI - Hypoxic-ischemic brain injury in infants with congenital heart disease dying after cardiac surgery. AB - Cardiac surgery for congenital heart disease is performed increasingly earlier in infancy, including in the neonatal period. With increased survival of infants, there is growing concern about the long-term neurological sequelae of hypoxic ischemic injury due to congenital heart disease itself prior to surgery, corrective surgery with the use of low-flow cardiopulmonary bypass (CPB) and/or deep hypothermic circulatory arrest (DHCA), and/or unstable hemodynamic factors postoperatively. In analyzing the neuropathology of 38 infants dying after cardiac surgery, we tested a set of questions related to the severity and patterns of brain injury, CPB, DHCA, and age of the infants at the time of surgery. In all infants dying after cardiac surgery, irrespective of the modality, cerebral white matter damage [periventricular leukomalacia (PVL) or diffuse white matter gliosis] was the most significant lesion in terms of severity and incidence, followed by a spectrum of gray matter lesions. There was no significant association between the duration of deep hypothermic circulatory arrest and the degree of severity of overall brain injury, and the pattern of brain injury was similar irrespective of the modality of cardiac surgery. There was no significant association between the age at the time of surgery (neonatal versus postneonatal) and the severity of overall brain injury. The patterns of brain injury were not age-related in the limited time-frame analyzed, except that infants who developed acute PVL after both closed and DHCA/CPB surgery (14/38 infants, 34%) were significantly younger at death (median age 13.0 days) compared to unaffected infants (median age at death 42.5 days) (P=0.031). This observation suggests that neonatal (<30 postnatal days), but not postneonatal (>30 postnatal days), brains are at risk for acute PVL, and likely reflects the vulnerability of immature (pre-myelinating) white matter to hypoxia-ischemia. PMID- 16244892 TI - Nasal septal perforation in a patient with subclinical celiac disease: a possible new association. AB - A female patient presented with nasal septal perforation that did not respond to conventional therapeutic management. Later, because of a malabsorption problem in one of her children, she underwent analytic tests and distal duodenal biopsy, which revealed that she was suffering from subclinical/silent celiac disease. The treatment, a gluten-free diet, unexpectedly resulted in the cessation of the destructive nasal process. Four years later, the patient remains asymptomatic. Nasal septal perforation might constitute a new entity associated with celiac disease hitherto not described in the literature. PMID- 16244893 TI - Characterization of the ALP1 gene locus of Trichophyton tonsurans. AB - Trichophyton tonsurans is the primary etiologic agent of fungal infections in the pediatric population. Establishing techniques that facilitate strain discrimination offer the opportunity to investigate the relationship between fungal genotype, biochemical phenotype and disease presentation in the host. In the process of expanding efforts to elucidate intra-specific genetic variability in T. tonsurans, we have identified 2 genetic polymorphisms in the ALP1 gene: a fragment length polymorphism in the 5'UTR and a single SNP (G-->A) within the 3'UTR. Full sequence data revealed that the length polymorphism was constituted by a 16 bp repeat element, present in tandem from 3 to 6 times depending on the strain. Quantitative RT-PCR analysis demonstrated a clear association between the length polymorphism and ALP1 mRNA transcript levels. Not only do the sequence variations identified in this study increase our ability to discriminate T. tonsurans strains, but they also reveal the presence of a genetic variation with functional consequences at the transcript level that may play a role in regulating disease severity. PMID- 16244894 TI - Farmer's lung disease and microbiological composition of hay: a case-control study. AB - Previous studies performed in France have suggested that handling hay contaminated with high amounts of moulds, and especially Absidia corymbifera and Eurotium amstelodami, may favour farmer's lung disease. The circumstances favouring farmer's lung disease and the distinctive microbiological composition of hay samples that provoke attacks need to be specified. We present a case control study which investigates the agricultural practices and the microbiological composition of hay handled in patients with farmer's lung disease as compared to those of a representative control population. Ten cases identified the hay they were handling at the onset of symptoms. The location, type of farm and working conditions were similar to those of the control farms. Conversely, the microbiological composition of hay differed, with significantly higher amounts of E. amstelodami (P < 0.01), A. corymbifera (P = 0.003), mesophilic Streptomyces (P < 0.01), thermophilic Streptomyces (P < 0.01) and Saccharomonospora viridis (P < 0.01) than in the control population. Our results demonstrate that hay identified by patients as having a harmful effect is characterized by a higher total amount of microorganisms, notably five microorganisms that seem discriminative. Mean concentrations are 2- to 115-fold higher in hay suspected to cause symptoms than in hay from a representative panel of farms. Handling hay with high amounts of these five microorganisms constitutes a risk factor for farmer's lung disease that should be considered for the development of prophylactic measures. PMID- 16244895 TI - Analysis of dermatomycoses in Lanzhou district of northwestern China. AB - The skin mycoses, perticularly dermatophytoses, in Lanzhou district, Northwestern China, was investigated during July 2002-June 2003. The specimens from patients suspected of having dermatomycoses were examined microscopically in KOH preparations and cultured on Sabouraud dextrose agar (SDA). Among 1443 suspected cases, 594 were KOH positive and 221 cultures of fungi were isolated. The most frequently isolated fungi were Trichophyton rubrum (43.9%) Trichophyton mentagrophytes (29.4%) and Candida species (14.0%). The frequency of tinea pedis, onychomycosis and tinea manuum were 38.7, 27.8 and 13.5%, respectively. In Lanzhou district, tinea pedis is the most commonly seen dermatophytoses, and T. rubrum is the most frequent etiologic agent. PMID- 16244896 TI - Mating-type distribution and fertility status in Magnaporthe grisea populations from Argentina. AB - Isolates of Magnaporthe grisea causing gray leaf spot on rice were collected in Argentina and analyzed for mating distribution and fertility. One hundred and twenty-five isolates of M. grisea were collected from rice plants between 2000 and 2003. Each isolate was tested for mating type through a polymerase chain reaction based assay. All M. grisea isolates from Argentina belonged to a single mating type, MAT1.1. The fertility status of isolates was determined using controlled crosses in vitro, pairing each isolate with GUY11 and KA9 (MAT1.2 standard hermaphroditic testers). Production of perithecia was scarce among isolates of the blast pathogen since a low percentage of them (7.2%) developed perithecia with only one of the fertile tester (KA9); all crosses failed with the other tester strain. Asci and ascospores were not observed. The presence of only one mating type and the absence of female fertile isolates indicate that sexual reproduction is rare or absent in M. grisea populations associated with rice in Argentina. PMID- 16244897 TI - Evaluation of Fusarium solani hyphae and conidia susceptibility to amphotericin B and itraconazole: study of a clinical case. AB - Fusarium species are hyaline moulds belonging to the hyalohyphomycosis group that are usually found in the soil and plants. This organism has emerged as a cause of disseminated invasive disease. The correlation between in vitro value and clinical efficacy is low and many patients remain unresponsive to treatment despite in vitro susceptibility. We determined growth control for Fusarium solani using the BioCell-Tracer system that measures the growth rate of a single fungal hypha, and the effect of different concentrations of amphotericin B and itraconazole. The MIC for these two drugs was also determined by a broth microdilution technique, using RPMI 1640. Different MICs for amphotericin B were obtained by the two different methods. This paper describes a case of infection due to Fusarium solani in an allogeneic bone marrow transplanted patient, the microbiological diagnostic, antifungal susceptibility tests for conidia and hypha and clinical correlation. PMID- 16244898 TI - Reduction of deoxynivalenol in barley by treatment with aqueous sodium carbonate and heat. AB - Naturally contaminated lots of Canadian barley containing either 18.4 or 4.3 microg/g deoxynivalenol (DON) were heated at 80 degrees C, with small amounts of water or 1 M sodium carbonate solution to study the rate of DON reduction. Samples were heated in sealed polypropylene containers for periods of up to 8 days. In the 18.4 microg/g DON barley, rapid reductions were observed: with no solutions added, DON declined to 14.7 microg/g after 1 day, and to 4.9 microg/g after 8 days solely due to heat; with water at 10 mL/100 g barley, DON levels reached 3.7 microg/g after 8 days; with 1 M sodium carbonate solution added at 10 mL/100 g barley, DON declined to 4.7 microg/g after 1 day, and to 0.4 microg/g after 8 days; with 20 mL/100 g barley, DON declined to 1.4 microg/g after 1 day and to near-zero levels after 8 days. In the 4.3 microg/g DON barley, more gradual reductions were evident: with no solutions added, DON declined to 2.9 microg/g after 8 days solely due to heat; with water at 10 mL/100 g barley, DON levels reached 2.3 microg/g after 8 days; with 1 M sodium carbonate solution added at 10 mL/100 g barley, DON declined to 2.7 microg/g after 1 day, and to near-zero levels after 8 days; with 20 mL/100 g barley, DON declined to 1.4 microg/g after 1 day and to near-zero levels after 3, 5 and 8 days. PMID- 16244899 TI - Differential expression of chitin synthase (CHS) and glucan synthase (FKS) genes correlates with the formation of a modified, thinner cell wall in in vivo produced Beauveria bassiana cells. AB - During infection (in vivo), the entomopathogenic fungus Beauveria bassiana produces yeast-like cells that are surrounded by modified cell walls. These modifications have been related to the fungus ability to limit recognition by the host defense system. The composition of the in vivo cell wall was analyzed using a combination of cytochemical and molecular techniques. The in vivo cell walls still contained both chitin and 1,3-beta-glucan, but they were significantly thinner than in vitro cell walls (50-60 nm versus 100-160 nm, respectively). The difference in cell wall thickness was correlated with transcriptional regulation of cell wall-related genes: quantitative RT-PCR reactions demonstrated that B. bassiana chitin synthase (CHS) and glucan synthase (FKS) genes are down regulated in vivo. These analyses indicate that in vivo-triggered phenotypic modifications, including cell wall adjustments, are controlled by molecular mechanisms that include regulation of gene expression at the transcriptional level. PMID- 16244900 TI - Pathogenicity of the entomopathogenic fungus, Lecanicillium muscarium, against the sweetpotato whitefly Bemisia tabaci under laboratory and glasshouse conditions. AB - The potential for using the entomopathogenic fungus Lecanicillium muscarium to control the sweetpotato whitefly, Bemisia tabaci has been established in the laboratory by other studies. Laboratory studies however frequently overestimate the level of control achieved by biological control agents in the glasshouse. Before full-scale commercial or field development is considered, glasshouse trials are required to confirm laboratory results. Under both controlled laboratory and glasshouse conditions high mortality of second instar B. tabaci was recorded after application of L. muscarium. The potential of incorporating L. muscarium into integrated pest management strategies for the control of B. tabaci is discussed. PMID- 16244901 TI - Cultivation of entomopathogenic fungi for the search of antibacterial compounds. AB - Entomopathogenic fungi are a rich source of natural bioactive compounds. To establish cultivation conditions which facilitate the production of bioactive compounds and to select good genera among entomopathogenic fungi as the producer, 47 typical entomopathogenic fungi were tested for their ability to produce antibiotic activity. Thirty-eight strains (81%) and 30 strains (64%) of these fungi produced either anti-Bacillus compounds or anti-Staphylococcus compounds, respectively, indicating that the majority of the entomopathogenic fungi tested possessed the ability to produce antibacterial compounds. Using 9 representative strains (Aschersonia sp. HF724, Beauveria bassiana HF338, Cordyceps ramosopulvinata HF746, Metarhizium anisopliae HF293, Metarhizium flavoviride HF698, Nomuraea rileyi HF588, Paecilomyces fumosoroseus HF254, Paecilomyces tenuipes HF419, and Verticillium lecanii HF238), the cultivation conditions in liquid medium were surveyed with respect to the cultivation procedure and medium composition, particularly in terms of the presence or absence of insect-derived materials. At 26 degrees C, M. anisopliae HF293, N. rileyi HF588, and V. lecanii HF238 strains produced clear antibiotic activity against Bacillus and Saccharomyces, but only in the presence of insect-derived materials, suggesting that the production of antibacterial/antifungal compounds by entomopathogenic fungi is triggered by the presence of insect-derived materials. PMID- 16244905 TI - Structural, functional, and phylogenetic characterization of a large CBF gene family in barley. AB - CBFs are key regulators in the Arabidopsis cold signaling pathway. We used Hordeum vulgare (barley), an important crop and a diploid Triticeae model, to characterize the CBF family from a low temperature tolerant cereal. We report that barley contains a large CBF family consisting of at least 20 genes (HvCBFs) comprising three multigene phylogenetic groupings designated the HvCBF1-, HvCBF3 , and HvCBF4-subgroups. For the HvCBF1- and HvCBF3-subgroups, there are comparable levels of phylogenetic diversity among rice, a cold-sensitive cereal, and the cold-hardy Triticeae. For the HvCBF4-subgroup, while similar diversity levels are observed in the Triticeae, only a single ancestral rice member was identified. The barley CBFs share many functional characteristics with dicot CBFs, including a general primary domain structure, transcript accumulation in response to cold, specific binding to the CRT motif, and the capacity to induce cor gene expression when ectopically expressed in Arabidopsis. Individual HvCBF genes differed in response to abiotic stress types and in the response time frame, suggesting different sets of HvCBF genes are employed relative to particular stresses. HvCBFs specifically bound monocot and dicot cor gene CRT elements in vitro under both warm and cold conditions; however, binding of HvCBF4 subgroup members was cold dependent. The temperature-independent HvCBFs activated cor gene expression at warm temperatures in transgenic Arabidopsis, while the cold-dependent HvCBF4-subgroup members of three Triticeae species did not. These results suggest that in the Triticeae - as in Arabidopsis - members of the CBF gene family function as fundamental components of the winter hardiness regulon. PMID- 16244906 TI - NADK2, an Arabidopsis chloroplastic NAD kinase, plays a vital role in both chlorophyll synthesis and chloroplast protection. AB - As one of terminal electron acceptors in photosynthetic electron transport chain, NADP receives electron and H(+) to synthesize NADPH, an important reducing energy in chlorophyll synthesis and Calvin cycle. NAD kinase (NADK), the catalyzing enzyme for the de novo synthesis of NADP from substrates NAD and ATP, may play an important role in the synthesis of NADPH. NADK activity has been observed in different sub-cellular fractions of mitochondria, chloroplast, and cytoplasm. Recently, two distinct NADK isoforms (NADK1 and NADK2) have been identified in Arabidopsis. However, the physiological roles of NADKs remain unclear. In present study, we investigated the physiological role of Arabidiposis NADK2. Sub-cellular localization of the NADK2-GFP fusion protein indicated that the NADK2 protein was localized in the chloroplast. The NADK2 knock out mutant (nadk2) showed obvious growth inhibition and smaller rosette leaves with a pale yellow color. Parallel to the reduced chlorophyll content, the expression levels of two POR genes, encoding key enzymes in chlorophyll synthesis, were down regulated in the nadk2 plants. The nadk2 plants also displayed hypersensitivity to environmental stresses provoking oxidative stress, such as UVB, drought, heat shock and salinity. These results suggest that NADK2 may be a chloroplast NAD kinase and play a vital role in chlorophyll synthesis and chloroplast protection against oxidative damage. PMID- 16244907 TI - Identification of membrane-associated proteins regulated by the arbuscular mycorrhizal symbiosis. AB - A sub-cellular proteomic approach was carried out to monitor membrane-associated protein modifications in response to the arbuscular mycorrhizal (AM) symbiosis. Membrane proteins were extracted from Medicago truncatula roots either inoculated or not with the AM fungus Glomus intraradices. Comparative two-dimensional electrophoresis revealed that 36 spots were differentially displayed in response to the fungal colonization including 15 proteins induced, 3 up-regulated and 18 down-regulated. Among them, seven proteins were found to be commonly down regulated in AM-colonized and phosphate-fertilized roots. Twenty-five spots out of the 36 of interest could be identified by matrix assisted laser desorption/ionisation-time of flight and/or tandem mass spectrometry analyses. Excepting an acid phosphatase and a lectin, none of them was previously reported as being regulated during AM symbiosis. In addition, this proteomic approach allowed us for the first time to identify AM fungal proteins in planta. PMID- 16244908 TI - Expression patterns of purple acid phosphatase genes in Arabidopsis organs and functional analysis of AtPAP23 predominantly transcribed in flower. AB - Purple acid phosphatases (PAPs) are metallo-phosphoesterases. Their expression and function have not been systematically investigated in higher plants. In this work, we compared the transcript levels of 28 Arabidopsis PAP (AtPAP) genes in five Arabidopsis organs. The 28 members, although differed in their expression patterns in vegetative organs, were all transcribed in flower. Furthermore, the transcription of seven members (AtPAPs 6, 11, 14, 19, 23, 24 and 25) occurred predominantly in the flower. To begin dissecting the role of AtPAP genes in flower development, further expression and functional analyses were conducted using AtPAP23. Histochemical staining of transgenic plants expressing AtPAP23 promoter-beta-glucuronidase (GUS) gene construct revealed that AtPAP23 transcription was strong in flower apical meristems, but became restricted to petals and anther filaments in fully developed flower. A GST (glutathione S transferase) fusion protein of AtPAP23 (GST:AtPAP23) was expressed in bacterial cells, and was found to contain significant amounts of Fe and Mn (whereas the control GST protein contained none). In biochemical tests, GST:AtPAP23 showed typical acid phosphatase activities. The fusion protein was also highly active on phosphoserine, but not phosphotyrosine. Despite its highly specific expression pattern and the demonstrated biochemical function of its protein product, the RNAi (RNA interference), T-DNA knock-out and overexpression lines of AtPAP23 were indistinguishable from wild type plants in the development of flower (or other organs). Interestingly, the Fe and Mn contents were found significantly increased in AtPAP23 overexpression lines, which may offer a new direction for further functional studies of AtPAPs in Arabidopsis. PMID- 16244909 TI - Active RNA silencing at low temperature indicates distinct pathways for antisense mediated gene-silencing in potato. AB - Previously, it was shown that low temperature (/=0.9, and the number of proteins detected increased to 345 when the minimum probability value was reduced to 0.5. Membrane-integral proteins of the reaction center, cytochrome b/c (1), light-harvesting and ATPase complexes were used as controls to assess how well this approach performs with hydrophobic proteins. New genes were identified, and tentatively designated as encoding photosynthesis-related proteins. We conclude that this approach is a powerful method to evaluate the possible existence of new photosynthesis-related proteins (and genes), although alternative methods are needed to evaluate the exact functions of newly discovered genes. PMID- 16245052 TI - Proteomics of chloroplast envelope membranes. AB - Proteomics is a very powerful approach to link the information contained in sequenced genomes, like Arabidopsis, to the functional knowledge provided by studies of plant cell compartments, such as chloroplast envelope membranes. This review summarizes the present state of proteomic analyses of highly purified spinach and Arabidopsis envelope membranes. Methods targeted towards the hydrophobic core of the envelope allow identifying new proteins, and especially new transport systems. Common features were identified among the known and newly identified putative envelope inner membrane transporters and were used to mine the complete Arabidopsis genome to establish a virtual plastid envelope integral protein database. Arabidopsis envelope membrane proteins were extracted using different methods, that is, chloroform/methanol extraction, alkaline or saline treatments, in order to retrieve as many proteins as possible, from the most to the less hydrophobic ones. Mass spectrometry analyses lead to the identification of more than 100 proteins. More than 50% of the identified proteins have functions known or very likely to be associated with the chloroplast envelope. These proteins are (a) involved in ion and metabolite transport, (b) components of the protein import machinery and (c) involved in chloroplast lipid metabolism. Some soluble proteins, like proteases, proteins involved in carbon metabolism or in responses to oxidative stress, were associated with envelope membranes. Almost one third of the newly identified proteins have no known function. The present stage of the work demonstrates that a combination of different proteomics approaches together with bioinformatics and the use of different biological models indeed provide a better understanding of chloroplast envelope biochemical machinery at the molecular level. PMID- 16245053 TI - Insights into the stress response and sulfur metabolism revealed by proteome analysis of a Chlorobium tepidum mutant lacking the Rubisco-like protein. AB - A significant fraction of the proteome of Chlorobium tepidum is altered in a mutant strain of the green sulfur bacterium C. tepidum (Omega::RLP) lacking the Rubisco-like protein (RLP). Additionally, a number of stress proteins display altered abundance or migration in strain Omega::RLP, including a thioredoxin, a putative Hsp20 family chaperonin, and GroEL. Changes in protein abundance are closely correlated to mRNA abundance in the case of two other stress proteins, a thiol-specific antioxidant protein homolog (Tsa/AhpC) and an iron only superoxide dismutase (Fe-SOD). Strain Omega::RLP is more resistant to hydrogen peroxide exposure than strain WT2321, providing evidence that the stress proteins are functional. Strain Omega::RLP is also defective in thiosulfate oxidation, but is able to oxidize sulfide as well as the wild-type strain. Based on studies with periplasm-enriched extracts of strain Omega::RLP, the loss of thiosulfate oxidation capability correlates with undetectable levels of the Sox Y protein, a component of the predicted thiosulfate oxidation complex. These results provide further indications that sulfur oxidation capacity and the response to stress are linked in C. tepidum, with the RLP playing a major role. PMID- 16245054 TI - The proteome of the chloroplast lumen of higher plants. AB - Recent research in proteomics of the higher plant chloroplast has achieved considerable progress and added to our knowledge of lumenal chloroplast proteins. This work shows that chloroplast lumen has its own specific proteome and may comprise as many as 80 proteins. Although the new map of the lumenal proteome provides a great deal of information, it also raises numerous questions because the physiological functions of most of the novel lumenal proteins are unknown. In this Minireview, we summarize the latest discoveries regarding lumenal proteins and present the currently known facts about the lumenal chloroplast proteome of higher plants. PMID- 16245055 TI - Thylakoid membrane proteomics. AB - Proteomics seeks to monitor the global complement of proteins within a cell or organism and accompanying plasticity with respect to development and environment. The proteome is dynamic, the product of current and past gene expression, countless protein-protein interactions and selective proteolytic systems. Consequently the snapshot that a proteomic measurement yields must be integrated into proteome flux; the flow of nutrients and energy through the protein pathways that catalyze and drive life. The thylakoid membrane proteome poses many technical challenges for proteomics. Integral membrane proteins present awkward physico-chemical properties and the abundant photosynthetic machinery conceals much less abundant and no less important proteins such as channels and transporters that control the interaction of stroma and lumen. Discussed here are contrasting approaches to thylakoid proteomics; 'shotgun' techniques that provide throughput benefits by cleaving proteins into smaller more-manageable peptide chunks versus intact protein techniques that provide more detailed and accurate pictures. A two-dimensional chromatography system directly interfaced to electrospray-ionization mass spectrometry has allowed the direct visualization of large reaction-center proteins (up to 83 kDa) from both Photosystems 1 and 2 providing an attractive avenue for characterization of thylakoid membrane proteomes under different conditions because of the ability to resolve molecular heterogeneity resulting from post-translational modifications such as phosphorylation and oxidation. A high-resolution spectrum of Bacteriorhodopsin recorded to an accuracy of 8 ppm using Fourier-transform mass spectrometry demonstrates the first application of this technique to intact polytopic integral membrane proteins. PMID- 16245058 TI - Contents volume 78 2003. PMID- 16245056 TI - Proteomic analysis of the cyanobacterium Anabaena sp. strain PCC7120 with two dimensional gel electrophoresis and amino-terminal sequencing. AB - A protein-gene linkage map of the cyanobacterium Anabaena sp. strain PCC7120 was successfully constructed for 123 relatively abundant proteins. The total proteins extracted from the cell were resolved by two-dimensional electrophoresis, and the amino-terminal sequences of the protein spots were determined. By comparing the determined amino-terminal sequences with the entire genome sequence, the putative translation initiation sites of 87 genes were successfully assigned on the genome. The elucidated sequence features surrounding the translation initiation sites were as follows: (1) GTG and TTG in addition to the ATG were used as rare initiation codons; (2) the core sequences (GAGG, GGAG and AGGA) of the Shine Dalgarno sequence were identified in the appropriate position preceding the 51 initiation sites (58.6%); (3) the nucleotides at the two regions, from -35 to 33, and from -19 to -17 (relative to the first nucleotide in the initiation codon) were preferentially adenines or thymines; (4) the nucleotides at the region from -14 to -8 were preferentially purines; (5) the nucleotide at position -1 was biased towards non-guanine (96.6%); (6) the nucleotide at the position +5 was preferentially cytosine (63.2%). It was evident that removal of the translation initiator methionine was dependent on the side-chain bulkiness of the penultimate amino acid residue. The predicted putative signal peptide sequences were also indicated. Besides confirming the existence of many predicted proteins, the data will serve as a starting point for the study of signals important in post-translational processing and nucleotide sequences important in the initiation of translation. PMID- 16245060 TI - Deciphering the 820 nm signal: redox state of donor side and quantum yield of Photosystem I in leaves. AB - By recording leaf transmittance at 820 nm and quantifying the photon flux density of far red light (FRL) absorbed by long-wavelength chlorophylls of Photosystem I (PS I), the oxidation kinetics of electron carriers on the PS I donor side was mathematically analyzed in sunflower (Helianthus annuus L.), tobacco (Nicotiana tabacum L.) and birch (Betula pendula Roth.) leaves. PS I donor side carriers were first oxidized under FRL, electrons were then allowed to accumulate on the PS I donor side during dark intervals of increasing length. After each dark interval the electrons were removed (titrated) by FRL. The kinetics of the 820 nm signal during the oxidation of the PS I donor side was modeled assuming redox equilibrium among the PS I donor pigment (P700), plastocyanin (PC), and cytochrome f plus Rieske FeS (Cyt f + FeS) pools, considering that the 820 nm signal originates from P700(+) and PC(+). The analysis yielded the pool sizes of P700, PC and (Cyt f + FeS) and associated redox equilibrium constants. PS I density varied between 0.6 and 1.4 mumol m(-2). PS II density (measured as O(2) evolution from a saturating single-turnover flash) ranged from 0.64 to 2.14 mumol m(-2). The average electron storage capacity was 1.96 (range 1.25 to 2.4) and 1.16 (range 0.6 to 1.7) for PC and (Cyt f + FeS), respectively, per P700. The best-fit electrochemical midpoint potential differences were 80 mV for the P700/PC and 25 mV for the PC/Cyt f equilibria at 22 degrees C. An algorithm relating the measured 820 nm signal to the redox states of individual PS I donor side electron carriers in leaves is presented. Applying this algorithm to the analysis of steady-state light response curves of net CO(2) fixation rate and 820 nm signal shows that the quantum yield of PS I decreases by about half due to acceptor side reduction at limiting light intensities before the donor side becomes oxidized at saturating intensities. Footnote: PMID- 16245061 TI - New fluorescence parameters for monitoring photosynthesis in plants. AB - Chlorophyll fluorescence measurements have a wide range of applications from basic understanding of photosynthesis functioning to plant environmental stress responses and direct assessments of plant health. The measured signal is the fluorescence intensity (expressed in relative units) and the most meaningful data are derived from the time dependent increase in fluorescence intensity achieved upon application of continuous bright light to a previously dark adapted sample. The fluorescence response changes over time and is termed the Kautsky curve or chlorophyll fluorescence transient. Recently, Strasser and Strasser (1995) formulated a group of fluorescence parameters, called the JIP-test, that quantify the stepwise flow of energy through Photosystem II, using input data from the fluorescence transient. The purpose of this study was to establish relationships between the biochemical reactions occurring in PS II and specific JIP-test parameters. This was approached using isolated systems that facilitated the addition of modifying agents, a PS II electron transport inhibitor, an electron acceptor and an uncoupler, whose effects on PS II activity are well documented in the literature. The alteration to PS II activity caused by each of these compounds could then be monitored through the JIP-test parameters and compared and contrasted with the literature. The known alteration in PS II activity of Chenopodium album atrazine resistant and sensitive biotypes was also used to gauge the effectiveness and sensitivity of the JIP-test. The information gained from the in vitro study was successfully applied to an in situ study. This is the first in a series of four papers. It shows that the trapping parameters of the JIP-test were most affected by illumination and that the reduction in trapping had a run-on effect to inhibit electron transport. When irradiance exposure proceeded to photoinhibition, the electron transport probability parameter was greatly reduced and dissipation significantly increased. These results illustrate the advantage of monitoring a number of fluorescence parameters over the use of just one, which is often the case when the F(V)/F(M) ratio is used. PMID- 16245062 TI - Characterization of the alterations of the chlorophyll a fluorescence induction curve after addition of Photosystem II inhibiting herbicides. AB - The effects of Photosystem II inhibiting herbicides, 3-(3,4-dichlorophenyl)-1,1 dimethylurea (diuron), atrazine and two novel 2-benzylamino-1,3,5-triazine compounds, on photosynthetic oxygen evolution and chlorophyll a fluorescence induction were measured in thylakoids isolated from Chenopodium album (wild type and atrazine-resistant plants) and cyanobacterial intact cells. The resistant plants have a mutation of serine for glycine at position 264 of the D1 protein. Diuron and two members of a novel class of 2-benzylamino-1,3,5-triazine compounds were almost as active in wild-type as in atrazine-resistant thylakoids, indicating that the benzylamino substitution in the novel triazines may be important for the lack of resistance in these atrazine-resistant plants. The inhibition by the herbicides of oxygen evolution in the cyanobacteria was somewhat lower than in the thylakoids of Chenopodium album wild type, probably caused by a slower uptake in the intact cells. The so-called OJIP fluorescence induction curve was measured during a one second light pulse in the absence and in the presence of high concentrations of the four herbicides. In the presence of a herbicide we observed an increase of the initial fluorescence at the origin (Fo'), a higher J level, and a decreased steady state at its P level (Fp). The increase to Fo' and the decreased leveling Fp are discussed. After dark adaptation about 25% of the reaction centers are in the S(0) state of the oxygen evolving complex with an electron on the secondary electron accepting quinone, Q(B). The addition of a herbicide causes a transfer of the electron on Q(B) to the primary quinone acceptor, Q(A), and displacement of Q(B) by the herbicide; the reduced Q(A) leads to a higher Fo'. The decrease of Fp in the presence of the herbicides is suggested to be caused by inhibition of the photo-electrochemical stimulation of the fluorescence yield. PMID- 16245063 TI - Phytol as one of the determinants of chlorophyll interactions in solution. AB - Optical absorption and fluorescence parameters of chlorophyll a and the phytol free chlorophyllide a, as well as of their Mg-depleted derivatives, were compared in a series of organic solvents. In contrast to prevailing opinion, the spectral properties of chlorophyll are not indifferent to the removal of phytol. The electronic absorption spectra of chlorophyll a and chlorophyllide a differ and display a different dependence on the nature of the solvent, which cannot be explained solely by the location of a charged carboxylic group in the proximity of the pi- electron system. In fact, measurements in media of varying basicity show that deprotonation of the free carboxylic group in chlorophyllide, i.e., the presence of a negative point charge near the macrocycle, has no effect on pigment absorption spectra. Analysis of the solvent effect on the Q(Y) energies in terms of solvent polarity reveals that the phytyl moiety perturbs the spectral features of chlorophyll, mainly due to its interactions with the pigment solvation shell. The phytyl residue might also be thus partly involved in controlling the central metal ligation in chlorophylls. This influence of phytol on the spectral features of chlorophyll should be taken into account when comparing the spectra in solution with various spectral forms of chlorophyll in vivo. PMID- 16245064 TI - Regulation of the expression of ribulose-1,5-bisphosphate carboxylase/oxygenase (EC 4.1.1.39) in a cyanobacterium, Synechococcus PCC7942. AB - When cyanobacterium cells are grown under extremely low CO(2) concentration, the number of carboxysomes, structures containing ribulose-bisphosphate carboxylase (Rubisco; EC 4.1.1.39), is known to increase. This suggests that Rubisco helps to regulate photosynthesis in cyanobacteria. However, no studies have been done on the changes of Rubisco content and activity in response to the extracellular CO(2) concentration, and no information is available on its effect on photosynthesis. To elucidate the relationship between the expression responses of Rubisco and extracellular CO(2), wild-type cells (Synechococcus PCC7942) and carboxysome-lacking cells were grown under various CO(2) concentrations, and Rubisco activity was determined. In both strains, Rubisco activity increased when the cells were grown under a CO(2) concentration around, or less than, K (1/2)(CO(2)) of photosynthesis. In carboxysome-lacking cells, Rubisco activity increased five to six times at most, and a simultaneous increase in the rate of photosynthesis was observed. These results suggest that stimulation of expression of Rubisco occurs to compensate for the decrease in the rate of photosynthesis under CO(2)-limited conditions. PMID- 16245065 TI - Photosynthetic and respiratory electron transport in the alkaliphilic cyanobacterium Arthrospira (Spirulina) platensis. AB - Photosynthetic and respiratory electron transport and their interplay with ion transport have been studied in Arthrospira platensis, a filamentous alkaliphilic cyanobacterium living in hypersaline lakes. As typical for alkaliphiles, A. platensis apparently does not maintain an outward positive pH gradient at its plasma membrane. Accordingly, sodium extrusion occurs via an ATP-dependent primary sodium pump, in contrast to the Na(+)/H(+) antiport in most cyanobacteria. A. platensis is strongly dependent on sodium/bicarbonate symport for the uptake of inorganic carbon. Sodium extrusion in the presence of the Photosystem II inhibitor diuron indicates that a significant amount of ATP is supplied by cyclic electron transport around Photosystem I, the content of which in A. platensis is exceptionally high. Plastoquinol is oxidized by two parallel pathways, via the cytochrome b (6) f complex and a putative cytochrome bd complex, both of which are active in the light and in the dark. PMID- 16245066 TI - Retardation of photo-induced changes in Photosystem I submembrane particles by glycinebetaine and sucrose. AB - The protective role of co-solutes (glycinebetaine and sucrose) against photodamage in isolated Photosystem (PS) I submembrane particles illuminated (2000 muE m(-2) s(-1)) for various time periods at 4 degrees C was studied. The photochemical activity of PS I in terms of electron transport measured as oxygen uptake and P700 photooxidation was significantly protected. A photoinduced enhancement of oxygen uptake observed during the first hours of strong light illumination attributed to denaturation or dissociation of membrane-bound superoxide dismutase [Rajagopal et al. (2003) Photochem. Photobiol 77: 284-291] was also retarded by glycinebetaine and sucrose. Chlorophyll photobleaching resulting in a decrease of absorbance and a blue-shift of the absorbance maximum in the red was greatly delayed in the presence of co-solutes. This phenomenon was also observed in the chlorophyll-protein (CP) complexes of PS I particles exposed to strong illumination separated on non-denaturing poly-acrylamide gels. In this case, a decrease in the absorbance of the CP1b band coinciding with an increase of CP1a during the course of illumination and ascribed to oxidative cross-linking (Rajagopal et al. 2003) was also retarded. Our results, thus, clearly demonstrated for the first time that co-solutes could minimize the alteration of photochemical activity and chlorophyll-protein complexes against photodamage of PS I submembranes particles. PMID- 16245067 TI - Is dimethylsulfoxide a reliable solvent for extracting chlorophyll under field conditions? AB - Dimethylsulfoxide (DMSO) appears to be a reliable solvent for extracting chlorophyll (Chl), however, modification of standard methods may be necessary for some species under field conditions. We found that Chl extraction of whole leaf tissue with DMSO incubated at between 25 and 40 degrees C was generally similar to the 80% acetone method, except for one graminoid species that required maceration. There was little effect of incubation temperature or duration of incubation beyond 7 h on extraction efficiency, but DMSO extracts were less stable than acetone extracts during one week of cold storage, especially if they thawed during this period. Since Chl extraction methods may provide variable results, particularly in the field, studies using different solvents should be compared cautiously unless specific methods have been calibrated. PMID- 16245068 TI - Estradiol induces PKA activation through the putative membrane receptor in the living hippocampal neuron. AB - For the past decade, estrogen actions rapid in onset, short in duration through the putative membrane estrogen receptor have attracted considerable attention. Nevertheless, there is so far limited evidence for estrogenic nongenomic regulation of the adenyl cyclase-cAMP-PKA system, especially in the brain. The present study reconfirms that 17beta-estradiol induces membrane-mediated PKA activation with a short latency in a living hippocampal neural cell with use of BSA-conjugated beta-estradiol which does not reach the nuclear estrogen receptor following my previous publication. PMID- 16245069 TI - Effects of nicotine on neurogenesis and plasticity of hippocampal neurons. AB - To study nicotine's effects on neurogenesis in the dentate gyrus of the hippocampus, nicotine was injected intraperitoneally into adult rats. After sacrificing, the hippocampal formation was processed for immunohistochemical staining of PSA-NCAM, NeuN and GFAP. Nicotine decreased numbers of PSA-NCAM(+) and NeuN(+) cells dose-dependently. PMID- 16245070 TI - Amygdala responsiveness is modulated by tryptophan hydroxylase-2 gene variation. AB - The tryptophan hydroxylase-2 gene (TPH2) codes for the enzyme of serotonin (5-HT) synthesis in the brain and variation of TPH2 has been implicated in disorders of emotion regulation. Here, we used functional magnetic resonance imaging (fMRI) to demonstrate that a potentially functional variant of TPH2 modulates amygdala responsiveness to emotional stimuli of both negative and positive valence. PMID- 16245071 TI - High activity of acid sphingomyelinase in major depression. AB - Acid sphingomyelinase (A-SMase) and its reaction product ceramide may play a role in the pathophysiology of depressive disorders and in the therapeutic action of antidepressive drugs. In a prospective case-control study, A-SMase activity was measured in peripheral blood mononuclear cells of 17 patients with a major depressive episode who were free of antidepressant drug therapy for at least 10 days and 8 healthy volunteers. In the patient group, A-SMase activity was correlated to the score (n=17, r=0.64, P=0.005). The patient group exhibited higher A-SMase activity compared to healthy volunteers (T=2.09, df=21.33, P<0.05). In addition, we demonstrate that the antidepressants imipramine and amitriptyline induce a long-term reduction of the activity of A-SMase in cultured cells. PMID- 16245072 TI - Cognitive impairment and its association with homocysteine plasma levels in females with eating disorders - findings from the HEaD-study. AB - Higher plasma homocysteine levels have been found in females with anorexia nervosa. Furthermore, elevated homocysteine levels are associated with cognitive decline in dementia and healthy elderly people. Aim of this prospective study was to investigate a possible association between homocysteine serum levels and Clinically well known cognitive deficits in females with eating disorders. We found that moderately elevated plasma homocysteine levels were associated with normal short- and long-term verbal memory while normal plasma homocysteine levels were associated with poorer memory performance in 14 females with anorexia nervosa and 12 females with bulimia nervosa (logistic forward regression Wald chi(2)=8.566, OR=24.75, CI 2.89 - 212.23, P=0.003). These results indicate that under the special circumstances of eating disorders elevated homocysteine levels improve memory signaling possibly by facilitating long-term potentiation. PMID- 16245074 TI - Interleukin 6 and 12, alanine aminotransferase activity, and HCV viral load in children with chronic hepatitis C treated with interferon and ribavirin. AB - The response to viral infections is mediated through the co-operation of cellular and humoral mechanisms. The aim of this study was to seek the correlation between IL-6 and IL-12 level, HCV viral load, ALT activity during the 48-week treatment with interferon-alpha-2b (IFN-alpha-2b) combined with ribavirin in children with diagnosed CHC and to search their influence on positive response to treatment. The group of 27 children with CHC was enrolled into this study. The children were treated with interferon-alpha and ribavirin in the course of 48-week therapy. The results show that both ALT activity and the viral load at the time of implementation of treatment with IFN-alpha and ribavirin is an important prognostic tool when treating children. It has been shown that the levels of IL-6 do not bear any significant prognostic importance to the implemented therapy, yet the increase of IL-12 levels in the 24th week of the treatment may be of prognostic value and may point out the possible elimination of HCV-RNA. PMID- 16245073 TI - Hypothesis: the humoral immune response to oral bacteria provides a stimulus for the development of rheumatoid arthritis. AB - Rheumatoid arthritis (RA) and adult periodontitis share common pathogenetic mechanisms and immunologic and pathological findings. One oral pathogen strongly implicated in the pathogenesis of periodontal disease, Porphyromonas gingivalis, possesses a unique microbial enzyme, peptidylarginine deiminase (PAD), the human equivalent of which has been identified as a susceptibility factor for RA. We suggest that individuals predisposed to periodontal infection are exposed to antigens generated by PAD, with de-iminated fibrin as a likely candidate, which become systemic immunogens and lead to intraarticular inflammation. PAD engendered antigens lead to production of rheumatoid factor-containing immune complexes and provoke local inflammation, both in gingiva and synovium via Fc and C5a receptors. PMID- 16245075 TI - Effects of granulocyte colony-stimulating factor (G-CSF) and neutrophil elastase inhibitor (ONO-5046) on acid-induced lung injury in rats. AB - It has been suggested that neutrophils play an important role in acid-aspirated lung injury. We examined the effects of the high dose of granulocyte-colony stimulating factor (G-CSF), which is capable of increasing peripheral neutrophils, and a specific neutrophil elastase inhibitor (ONO-5046) on acid lung injury in rats. Animals were anesthetized and normal saline (NS, 2 mL kg(-1)) or hydrochloric acid (HCl, 0.1 N 2 mL kg(-1)) was then instilled into trachea. Thirty minutes before HCl instillation, G-CSF (150 microg kg(-1)) was injected subcutaneously or ONO-5046 (10 mg kg(-1) h(-1)) was infused continuously into the right jugular vein. Animals were ventilated during the experiments. Five hours after HCl or NS instillation, bronchoalveolar lavage fluid (BALF) and lung tissue samples were obtained. Total nuclear cell count, absorbance, albumin, tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, cytokine-induced neutrophil chemoattractant (CINC), neutrophil elastase in BALF, wet-to-dry (W/D) ratio were measured. HCl aspiration markedly increased these values in BALF and W/D ratio. Both ONO-5046 and G-CSF attenuated the parameters increased by acid-induced lung injury in rats. The data suggests that neutrophils play an important role in acid induced lung injury. However, high-dose G-CSF does not exacerbate acid-aspirated lung injury in rats, although this agent causes an increase in peripheral neutrophils. PMID- 16245076 TI - A bactericidal cecropin-A peptide with a stabilized alpha-helical structure possess an increased killing capacity but no proinflammatory activity. AB - Antibacterial peptides are part of the innate immune system in a variety of different species including humans. Some of these peptides have also been shown to have effects on immune competent cells such as professional phagocytes. We have recently shown that a cecropin-like peptide from Helicobacter pylori, Hp(2 20), in addition to being bactericidal possesses proinflammatory effects and can recruit and activate neutrophils as well as monocytes. It is well established that cecropins have the ability to adopt amphipathic alpha-helices, which is thought to be required for their bactericidal activity. In this study we show the same structural requirements for Hp(2-20). Breaking the helical structure of Hp(2 20) reduced the antibacterial effect and abolished its proinflammatory activity. A C-terminal truncated cecropin A peptide that highly resembles Hp(2-20) failed to activate neutrophils and computer-based structural simulations revealed a difference between the two peptides in the stability of their helical structures. A hybrid peptide with amino acid substitutions stabilizing the alpha-helical structure of the truncated cecropin A peptide did not introduce any proinflammatory activity; the bactericidal activity was, however, increased. We thus conclude that the proinflammatory effect of Hp(2-20) is a unique sequence specific feature of the peptide and the ability to adopt a stable amphipathic helix is a necessary but not sufficient criterion for the functional dualism of the peptide. PMID- 16245077 TI - Enteropathogenic Escherichia coli outer membrane proteins induce iNOS by activation of NF-kappaB and MAP kinases. AB - Enteropathogenic Escherichia coli (EPEC) infects the human intestinal epithelium and is a major cause of infantile diarrhea in developing countries. Nitric oxide (NO) is an important modulator of intestinal inflammatory response. The aim of the present study was to investigate whether EPEC outer membrane proteins (OMPs) up regulate epithelial cell expression of inducible nitric oxide synthase (iNOS) and to examine the role of NF-kappaB and MAP kinases (MAPK) on nitrite production. iNOS mRNA expression was assessed by RT-PCR. Nitrite levels were measured by Griess reaction. NF-kappaB activation by OMPs was evaluated by EMSA and immunoblotting was done to detect MAPK activation. EPEC OMP up regulated iNOS, induced nitrite production and NF-kappaB and MAPK were activated in caco-2 cells. The nitrite levels decreased when NF-kappaB and MAPK inhibitors were used. Thus, EPEC OMPs induce iNOS expression and NO production through activation of NF kappaB and MAPK. PMID- 16245078 TI - Inflammatory granulocytes decrease subcutaneous growth of melanoma in mice. AB - Growing melanomas invade the subcutaneous tissues. We have compared the size of tumors implanted in the subcutaneous cavities of C57BL/6 mice where inflammatory reactions were induced before the injection of 5 x 10(5) melanoma cells (B16F10 cell line). Granulocytic inflammation of the subcutaneous cavities resulted in a significant decrease in the growth of the implanted melanomas, whereas monocytic inflammation had no effect on tumor growth. We conclude that granulocytes, but not monocytes/macrophages, have anti-tumor action on melanoma that invade the subcutaneous tissues. PMID- 16245079 TI - ICAM-1, ICAM-2 and ICAM-3 in the sera of patients with idiopathic pulmonary fibrosis. AB - In order to test the serum levels of ICAM-1, ICAM-2 and ICAM-3 in patients with idiopathic pulmonary fibrosis (IPF), twenty patients with IPF and eleven with secondary interstitial fibrosis (SIF), as well as forty healthy volunteers (HV) were studied. Serum intracellular adhesion molecules (ICAM) 1, 2 and 3 were assessed by ELISA. Functional respiratory tests, which included spirometry and lung diffusing capacity were simultaneously performed. Median values of serum ICAM-1 and ICAM-2 were higher in the patients' than in the healthy volunteers' (HV) group: IPF group: 946.60 ng/ml and 400.14 ng/ml; SIF group: 901.58 ng/ml and 378.27 ng/ml; HV group: 308.40 ng/ml and 217.55 ng/ml, respectively (p<0.05). ICAM-3 serum levels were equal between the three groups. ICAM-2 negatively correlated to DLCO values. (p<0.005). It can be concluded that ICAM 1 and 2 are elevated in the sera of patients with pulmonary fibrosis. ICAM-2 might be associated with a more impaired clinical status. PMID- 16245080 TI - Effect of organic cosolvent on kinetic resolution of tert-leucine by penicillin G acylase from Kluyvera citrophila. AB - Penicillin G acylase (PGA) from Kluyvera citrophila immobilized on Amberzyml was used for enantioselective hydrolysis of N-phenylacetylated-DL-tert-leucine (N Phac-DL-Tle) to produce L-tert-leucine (L-Tle). The effects of various organic cosolvents on hydrolysis of N-Phac-DL-Tle have been investigated in aqueous cosolvent medium. It was founded that the rate of PGA-catalyzed reaction was significantly affected by the presence of 2% (v/v) organic cosolvent concentration. The initial rate fell with increasing logP of the cosolvent, but for logP values less than -0.24 the rate was faster than in purely aqueous medium. Additionally, the relative rate increases with the increase of dielectric constant (epsilon) of organic cosolvents. The yields of L-Tle in all aqueous cosolvent systems were above 95% with the enantiomeric excess (ee) of >99%. PMID- 16245081 TI - A comparative structural and functional analysis of cyanobacterial plastocyanin and cytochrome c (6) as alternative electron donors to Photosystem I. AB - Plastocyanin and cytochrome c (6) are two soluble metalloproteins that act as alternative electron carriers between the membrane-embedded complexes cytochromes b (6) f and Photosystem I. Despite plastocyanin and cytochrome c (6) differing in the nature of their redox center (one is a copper protein, the other is a heme protein) and folding pattern (one is a beta-barrel, the other consists of alpha helices), they are exchangeable in green algae and cyanobacteria. In fact, the two proteins share a number of structural similarities that allow them to interact with the same membrane complexes in a similar way. The kinetic and thermodynamic analysis of Photosystem I reduction by plastocyanin and cytochrome c (6) reveals that the same factors govern the reaction mechanism within the same organism, but differ from one another. In cyanobacteria, in particular, the electrostatic and hydrophobic interactions between Photosystem I and its electron donors have been analyzed using the wild-type protein species and site-directed mutants. A number of residues similarly conserved in the two proteins have been shown to be critical for the electron transfer reaction. Cytochrome c (6) does contain two functional areas that are equivalent to those previously described in plastocyanin: one is a hydrophobic patch for electron transfer (site 1), and the other is an electrically charged area for complex formation (site 2). Each cyanobacterial protein contains just one arginyl residue, similarly located between sites 1 and 2, that is essential for the redox interaction with Photosystem I. PMID- 16245082 TI - Analysis of light and CO(2) regulation in Chlamydomonas reinhardtii using genome wide approaches. AB - Over the past decade new technologies have been developed to elucidate ways in which cells acclimate to environmental change. Many of these techniques have allowed the identification of specific transcripts that change in abundance in response to particular environmental stimuli; such transcripts represent genes that are potentially differentially regulated. Two techniques that foster identification of differentially regulated genes are differential display and expression profiling using high density DNA microarrays. The former technology amplifies cDNA fragments from mRNAs that differentially accumulate under specific environmental conditions, while the latter provides a more global view of changes in gene expression in response to environmental stimuli. Coupling these technologies with the analysis of mutants aberrant for regulatory molecules that participate in acclimation processes will allow the identification of groups of genes controlled by specific regulatory elements. In this article we describe the use of differential display and DNA microarray profiling to examine environmentally-regulated gene expression. We also show specific experiments using the unicellular green alga Chlamydomonas reinhardtii, in which mRNA abundance is evaluated in response to both changing light and CO(2) conditions. PMID- 16245083 TI - The interaction of visible and UV-B light during photodamage and repair of Photosystem II. AB - In order to understand the mechanism of photodamage induced by solar radiation under natural conditions, we studied the interaction of visible and ultraviolet-B light in the inactivation and repair of the Photosystem II complex by using oxygen evolution and flash-induced chlorophyll fluorescence measurements. In isolated spinach thylakoids and Synechocystis 6803 cells, in which de novo protein synthesis is blocked by lincomycin, photodamage of Photosystem II by visible and UV-B light is characterized by linear semilogarithmic inactivation curves for both separate and combined illumination protocols. The extent of PS II inactivation obtained after combined illumination can be well simulated by assuming independent damaging events induced by visible and UV-B photons. In intact Synechocystis cells capable of protein repair, simultaneous illumination by visible and UV-B light impairs Photosystem II activity to a smaller extent than expected from the independent damaging events. This protective effect is pronounced at low visible light (130 muE m(-2) s(-1)), but becomes negligible at high intensities (1300 muE m(-2) s(-1)). Exposure of intact Synechocystis 6803 cells to direct sunlight leads to a rapid inactivation of PS II, accompanied by the accumulation of donor side inhibited centers. This phenomenon, which shows the impairment of the manganese cluster of water oxidation was not observed when the ultraviolet components of sunlight were filtered out. We conclude that visible and UV-B photons inactivate PS II via non-interacting mechanisms, which affect different target sites. In intact cells, the two spectral regions do interact, and results in synergistically enhanced protein repair capacity when UV B radiation is accompanied by low intensity visible light, which provides protection against photodamage. However, this ameliorating effect becomes insignificant at high light intensities characteristic of direct sunlight. PMID- 16245084 TI - The action of a range of supplementary ultraviolet (UV) wavelengths on photosynthesis in Brassica napus L. in the natural environment: effects on PS II, CO(2) assimilation and level of chloroplast proteins. AB - The effects of different wavebands of UV radiation on photosynthesis and the expression and abundance of photosynthetic proteins in oilseed rape (Brassica napus L. cv. Rebel) were investigated. Plants were grown outdoors under natural radiation (52 degrees N, 0 degrees E) supplemented with six wavebands of UV radiation (0.4 Wm(-2)) between 313 nm and 356 nm. A control treatment was centred at 343 nm. Exposure to supplementary UV-A radiation (320-400 nm) had no significant effects, however UV-B radiation, centred at 313 nm, caused a marked reduction in photosynthesis. This decrease was related to a reduction in the initial carboxylation velocity of Rubisco which was further correlated with a large reduction in the expression and abundance of both large and small subunits of Rubisco. These results indicate a molecular mechanism behind UV-B induced reductions in photosynthesis per unit area in plants grown under field conditions. PMID- 16245085 TI - Low concentrations of NaHSO(3) increase cyclic photophosphorylation and photosynthesis in cyanobacterium Synechocystis PCC6803. AB - Application of NaHSO(3) solution at low concentrations (20-200 muM) to the culture medium enhanced photosynthetic oxygen evolution in cyanobacterium Synechocystis PCC6803 by more than 10%. The slow phase of ms-DLE was strengthened, showing that the transmembrane proton motive force related to photophosphorylation was enhanced. It was also observed that dry weight as well as ATP content under illuminated conditions were both increased after the treatment, indicating that low concentrations of NaHSO(3) could enhance the supply of ATP and thus increase biomass accumulation. In accord with the promotion in the photosynthetic oxygen evolution and ATP content, the transient increase in chlorophyll fluorescence after the termination of actinic light was increased; and meanwhile, the half-time of re-reduction of P700(+) in the presence of DCMU after a pulse light under background far-red light was shortened by approximately 30%, indicating that cyclic electron flow around PS I was accelerated by the treatment. Based on these results it is suggested that the increase in photosynthesis in Synechocystis PCC6803 caused by low concentrations of NaHSO(3) solution might be due to the stimulation of the cyclic electron flow around PS I and thus the increase in photophosphorylation. PMID- 16245086 TI - Time-resolved step-scan FTIR investigation on the primary donor of the reaction center from the green sulfur bacterium Chlorobium tepidum. AB - The vibrational properties of the primary donor P(840) in the reaction center (RC) of the green sulfur bacterium Chlorobium tepidum and its interactions with the surrounding protein environment have been investigated by Fourier transform infrared (FTIR) difference spectroscopy at cryogenic temperatures. By using the step-scan technique with a time resolution of 5 mus on RCs that had been depleted of the iron-sulfur electron acceptors, the formation and decay of the triplet state (3)P(840) have been followed in infrared for the first time. The (3)P(840)/P(840) FTIR difference spectrum is compared to the P(840) (+)/P(840) FTIR difference spectrum measured under identical conditions on untreated RCs and recorded with the same step-scan set-up. The latter P(840) (+)/P(840) difference spectrum is essentially the same as those measured under steady-state conditions using the more conventional continuous illumination method. Comparison of the (3)P(840)/P(840) and P(840) (+)/P(840) spectra provides unambiguous assignment of the vibration of the 9-keto C=O group(s) of P(840) at 1684 cm(-1) as the only common negative band in the two spectra. This frequency corresponds to carbonyl group(s) free from hydrogen bonding interactions. The obtained results are discussed in the framework of the structure and photochemistry of the primary donor P(840). PMID- 16245087 TI - A Synechococcus sp. PCC 7942 mutant with a higher tolerance towards bentazone. AB - In this article we describe the partial characterization of a Synechococcus sp. PCC 7942 mutant Mu1 with an enhanced resistance towards the herbicide bentazone (3-isopropyl-1H-2,1,3-benzothiadiazine-4(3H)-one 2,2-dioxide). The mutant was derived from a random mutagenesis with N-methyl-N'-nitro-N-nitrosoguanidine (NSG) and exhibited superior growth rates, pigment content and overall photosynthetic activities under regular growth conditions compared to wild type. Whereas Synechococcus PCC 7942 wild type showed significant photoinhibition, especially in the presence of lincomycin, Mu1 was much more robust. A comparative analysis of the content of several photosynthesis-associated proteins revealed that Mu1 had an increased expression of PsbO on mRNA and protein level and that PsbO is tightly bound to Photosystem II, relative to wild type. This result was substantiated by mass spectrometer measurements of photosynthetic water oxidation revealing a higher stability and integrity of the water oxidizing complex in Mu1 cells grown under regular or calcium deficient conditions. Therefore, our results give rise to the possibility that the overexpression of PsbO in mutant Mu1 confers resistance to reactive oxygen species (ROS) formed as a consequence of bentazone binding to the acceptor side of PS II. In addition, we observed a significantly higher tolerance towards bentazone in iron depleted wild type cells, conditions under which the IdiA protein becomes expressed in highly elevated amounts. As we have previously shown, IdiA preferentially protects the acceptor site of PS II against oxidative stress, especially under iron limitation. Thus, it is likely that IdiA due to its topology interferes with bentazone binding or protects PS II against ROS generated in the presence of bentazone. PMID- 16245088 TI - Quantification of photosynthetic gene expression in maize C(3) and C(4) tissues by real-time PCR. AB - Carbon assimilation in maize follows the C(4) mechanism. This requires the tissue specific and light-induced expression of a set of different genes involved in CO(2) fixation as well as adaptations in the leaf anatomy including a reduced distance between vascular bundles compared to C(3) plants. However, several maize tissues exist with larger bundle distances and there is significant evidence that CO(2) fixation follows the C(3) mechanism in these tissues. We isolated maize C(3) and C(4) tissues and quantified the accumulation of mRNAs encoding PEPC, ME, the small subunit of Rubisco, and PPDK. For this, primer systems for the specific and sensitive detection by real-time PCR were established. The observed patterns show the expected distribution for foliar leaf tissues. Also in total husk leaves, all transcripts under investigation were detected, albeit at a lower level. When mesophyll cells which are located distant from bundles were isolated from husk leaves, only accumulation of RbcS was observed. Comparing the expression of two genes encoding for isoenzymes of the small subunit of RbcS in the different tissues differential patterns of relative transcript abundance were observed. Transcripts for the DOF1 transcription factor involved in the activation of photosynthetic genes in maize were found in leaf tissues performing both C(4) and C(3) photosynthesis with highest accumulation levels in C(4) mesophyll cells, whereas the homologous DOF2 gene was not expressed in any of the investigated samples. The results provide novel insights into the regulation of C(3) and C(4) carbon fixation pathways in maize. PMID- 16245089 TI - The Calvin cycle revisited. AB - The sequence of reactions in the Calvin cycle, and the biochemical characteristics of the enzymes involved, have been known for some time. However, the extent to which any individual enzyme controls the rate of carbon fixation has been a long standing question. Over the last 10 years, antisense transgenic plants have been used as tools to address this and have revealed some unexpected findings about the Calvin cycle. It was shown that under a range of environmental conditions, the level of Rubisco protein had little impact on the control of carbon fixation. In addition, three of the four thioredoxin regulated enzymes, FBPase, PRKase and GAPDH, had negligible control of the cycle. Unexpectedly, non regulated enzymes catalysing reversible reactions, aldolase and transketolase, both exerted significant control over carbon flux. Furthermore, under a range of growth conditions SBPase was shown to have a significant level of control over the Calvin cycle. These data led to the hypothesis that increasing the amounts of these enzymes may lead to an increase in photosynthetic carbon assimilation. Remarkably, photosynthetic capacity and growth were increased in tobacco plants expressing a bifunctional SBPase/FBPase enzyme. Future work is discussed which will further our understanding of this complex and important pathway, particularly in relation to the mechanisms that regulate and co-ordinate enzyme activity. PMID- 16245090 TI - Rubisco activase - Rubisco's catalytic chaperone. AB - The current status of research on the structure, regulation, mechanism and importance of Rubisco activase is reviewed. The activase is now recognized to be a member of the AAA(+) family, whose members participate in macromolecular complexes that perform diverse chaperone-like functions. The conserved nucleotide binding domain of AAA(+) family members appears to have a common fold that when applied to the activase is generally consistent with previous site-directed mutagenesis studies of the activase. Regulation of the activase in species containing both isoforms can occur via redox changes in the carboxy-terminus of the larger isoform, mediated by thioredoxin-f, which alters the response of activase to the ratio of ADP to ATP in the stroma. Studies of Rubisco activation in transgenic Arabidopsis plants demonstrated that light modulation is dependent on redox regulation of the larger isoform, providing a model for the regulation in other species. Further insights into the mechanism of the activase have emerged from an analysis of the crystal structures of Rubisco conformational variants and the identification of Rubisco residues that confer specificity in its interaction with the activase. The physiological importance of the activase is reinforced by recent studies indicating that it plays a vital role in the response of photosynthesis to temperature. Rubisco activase is one of a new type of chaperone, which in this case functions to promote and maintain the catalytic activity of Rubisco. PMID- 16245091 TI - Action of UV and visible radiation on chlorophyll fluorescence from dark-adapted grape leaves (Vitis vinifera L.). AB - Grapevine plants (Vitis vinifera L. cv. Silvaner) were cultivated under shaded conditions in the absence of UV radiation in a greenhouse, and subsequently placed outdoors under filters transmitting natural radiation, or screening out the UV-B (280 to 315 nm), or screening out the UV-A (315 to 400 nm) and the UV-B spectral range. All conditions decreased maximum chlorophyll fluorescence (F(M)) and increased minimum chlorophyll fluorescence (F(0)) from dark-adapted leaves; however, with increasing UV, F(M) quenching was stimulated but increases in F(0) were reduced. The F(V)/F(M) ratio (where F(V)=F(M)-F(0)) was clearly reduced by visible radiation (VIS): UV-B caused a moderate extra-reduction in F(V)/F(M). Exposure of leaves (V. vinifera L. cv. Bacchus) to UV or VIS lamps quenched the F(M) to similar extents; further, UV-B doses comparable to the field, quenched F(0). A model was developed to describe how natural radiation intensities affect PS II and thereby change leaf fluorescence. Fitting theory to experiment was successful when the same F(M) yield for UV- and VIS-inactivated PS II was assumed, and for lower F(0) yields of UV- than for VIS-inactivated PS II. It is deduced, that natural UV can produce inactivated PS II exhibiting relatively high F(V)/F(M). The presence of UV-inactivated PS II is difficult to detect by measuring F(V)/F(M) in leaves. Hence, relative concentrations of intact PS II during outdoor exposure were derived from F(M). These concentrations, but not F(V)/F(M), correlated reasonably well with CO(2) gas exchange measurements. Consequently, PS II inhibition by natural UV could be a main factor for UV inhibition of photosynthesis. PMID- 16245092 TI - Study on the photo-generation of superoxide radicals in Photosystem II with EPR spin trapping techniques. AB - Direct EPR evidence of the photo-generation of superoxide radicals (O(2) (-.)) was obtained by using a novel spin trapping probe in spinach Photosystem II (PS II) membrane fragments. The production of O(2) (-.) was detected by following the formation of 5-diethoxyphosphoryl-5-methyl-1-pyrroline-N-oxide (DEPMPO) superoxide adducts (DEPMPO-OOH). The inhibition of O(2) (-.) formation by 3-(3,4 dichlorophenyl) -1,1-dimethylurea (DCMU) and the 77 K fluorescence spectrum indicated that O(2) (-.) were generated from PS II, not from PS I. The inhibition of O(2) (-.) formation by DCMU also suggested that O(2) (-.) were generated from the Q(B)binding site, not at a site prior to DCMU blockage. The extrinsic proteins and Mn are very important to eliminate O(2) (-.), showing that the oxygen-evolving system is involved in O(2) (-.) removal rather than production. PMID- 16245093 TI - The structure of the FMO protein from Chlorobium tepidum at 2.2 A resolution. AB - The bacteriochlorophyll protein, or FMO protein, from Chlorobium tepidum, which serves as a light-harvesting complex and directs light energy from the chlorosomes attached to the cell membrane to the reaction center has been crystallized in a new space group. The crystals belong to the cubic space group P4(3)32 and the structure has been refined to a resolution 2.2 A with a R factor of 19.7%. The electron density maps show that the structure is composed of two beta sheets that surround seven bacteriochlorophylls as previously reported (Li et al. (1997) J Mol Biol 271: 456-471). The availability of the new data allows a more accurate refinement of the pigment-protein complex including identification of bound solvent molecules. Several structural differences probably contribute to the observed spectroscopic differences between the FMO proteins from Cb. tepidum and Prosthecochloris aestuarii, including differences in the planarity of corresponding tetrapyrroles. A citrate molecule is found on the surface of each protein subunit of the trimer from Cb. tepidum. However, the citrate molecule is over 15 A from any bacteriochlorophyll. The presence of the citrate probably does not contribute to the function of the protein although it does contribute to the crystallization as it interacts with a crystallographically related trimer. Among the 236 water molecules found in the protein are four that appear to play a special role in the properties of bacteriochlorophyll 2, as this tetrapyrrole is coordinated by one of these water molecules and the waters form a hydrogen-bonded network that leads to the surface of the protein. PMID- 16245094 TI - Contrasting modes of regulation of PS II light utilization with changing irradiance in normal and psbS mutant leaves of Arabidopsis thaliana. AB - Complementary techniques of chlorophyll a fluorescence, steady state CO(2) exchange, and O(2) release during a multiple turnover flash were applied to compare responses to irradiance for leaves of wild type and psbS mutants. The latter included variants in which the psbS gene was deleted (npq4-1) or possessed a single point mutation (npq4-9). Nonphotochemical quenching (NPQ) was reduced by up to 80 and 50%, respectively, in these lines at high irradiance. Analysis of changes in steady-state fluorescence yields and quantum yield of linear electron transport in the context of the reversible radical pair model of Photosystem II (PS II) indicated that NPQ occurs by nonradiative deactivation of chlorophyll singlet states in normal leaves. In contrast, application of the same criteria together with the observed irreversibility of NPQ and decline in density of functional PS II reaction centers following excessive illumination indicated a change in reaction center properties for the psbS deletion phenotype (Npq4-1(-)). Specifically, PS II reaction centers in Npq4-1(-) convert to a photochemically inactive, yet strongly quenching, form in intense light. The possibility of formation of a carotenoid or chlorophyll cation quencher in the reaction center is discussed. Results for the point mutant phenotype (Npq4-9(-)) were intermediate to those of wild-type and Npq4-1(-). Furthermore, wild-type leaves exhibited a significant reversible increase in the PS II in vivo rate constant for photochemistry (k(P0)) in saturating compared to limiting light. Changes in k(P0) could not be accounted for in terms of a classic phosphorylation-dependent (state transition) mechanism. Changes in k(P0) may arise from alternate pigment protein conformations that alter the way excitons equilibrate among PS II chromophores. The lack of similar irradiance-dependent changes in k(P0) for the psbS mutants suggests a role for the PS II-S protein in the regulation of exciton distribution. PMID- 16245095 TI - Adaptation to iron deficiency: a comparison between the cyanobacterium Synechococcus elongatus PCC 7942 wild-type and a DpsA-free mutant. AB - To learn more about the adaptive response of Synechococcus elongatus PCC 7942 to iron starvation and the role of DpsA, presumably a protein protecting chromosomal DNA against oxidative damage, we performed a comparative analysis of S. elongatus PCC 7942 wild-type and a DpsA-free mutant, called K11. Relative to wild-type, the DpsA-free mutant had significantly higher amounts of phycocyanin and allophycocyanin, even upon iron limitation. While the Photosystem I activity in mutant K11 remained high under iron deficiency, the Photosystem II activity dropped severely with respect to wild-type. The DpsA content in wild-type was already fairly high under regular growth conditions and did not significantly increase under iron deficiency nor in the presence of 0.3 mM 2'2'-dipyridyl in iron-sufficient BG11 medium. Nevertheless, the absence of DpsA in K11 resulted in a significantly altered transcriptional/translational activity of genes known to be involved in adaptation to iron starvation. The amount of isiA/B transcript was about two-fold lower than in wild-type, resulting in a lower 77 K chlorophyll a fluorescence at 685 nm, implying a lower concentration of Photosystem I-IsiA supercomplexes. While in wild-type idiA, idiB, and irpA transcripts were highly up-regulated, hardly any were detectable in mutant K11 under iron limitation. The concentration of mapA transcript, however, was greatly increased in K11 compared to wild-type. Measurements of acridine yellow fluorescence with intact wild-type and K11 cells revealed that iron deficiency caused an increased contribution of cyclic electron transport to membrane energisation and ATP synthesis being in agreement with the formation of the Photosystem I-IsiA supercomplex. In addition, mutant K11 had a much higher respiratory activity compared to wild-type under iron limitation. PMID- 16245096 TI - The low-light reduction in the quantum yield of photosynthesis: potential errors and biases when calculating the maximum quantum yield. AB - Photosynthesis-irradiance (P-E) curves are widely used to describe photosynthetic efficiency and potential. Contemporary models assume maximal photosynthetic quantum yield (phi) at low irradiances. But P-E observations made with both oxygen evolution and carbon uptake techniques show that this is not always the case. Using new and published data in conjunction with modeling exercises, we demonstrate that regardless of the mechanism there can be reductions in phi at low irradiances that are not readily observable using conventional P-E analyses. We also show that analytical errors, such as inaccurate estimation of dark oxygen consumption or carbon uptake, can markedly affect the structure of phi-E curves with negligible effect on P-E curve structure. Whether from respiration ;corrections' or other mechanisms, these deviations in phi at low light levels from the maximum quantum yield of photosynthesis (phi(max)) can lead to significant errors (> 50%) in the estimation of the linear portion of the P-E curve and ultimately phi(max). Non-linear models of P-E, such as the rectangular hyperbola, quadratic, exponential and hyperbolic tangent that are commonly used to estimate the initial slope (alpha) of the P-E curve assume that phi is maximal at low light levels and therefore can err in the estimation of phi(max) when phi is reduced at low light levels. Using a diverse data set of 622 P-E curves with a total of 7623 points, we show that although model skills are high (r (2) = 0.96 +/- 0.05, 0.97 +/- 0.04, 0.97 +/- 0.04 and 0.97 +/- 0.04, respectively), a large fraction of the model-predicted phi(max) differ by greater than 10% from true phi(max) values (91%, 50%, 82% and 46%, respectively). Data from these observations and modeling exercises lead us to suggest that phi(max) be determined by directly estimating the true maximum of a phi-E curve rather than using the more conventional methodology employing the initial slope of the P-E curve. PMID- 16245097 TI - Celebrating the millennium - historical highlights of photosynthesis research. PMID- 16245098 TI - History of the word photosynthesis and evolution of its definition. AB - In 1893, Charles Barnes (1858-1910) proposed that the biological process for 'synthesis of complex carbon compounds out of carbonic acid, in the presence of chlorophyll, under the influence of light' should be designated as either 'photosyntax' or 'photosynthesis.' He preferred the word 'photosyntax,' but 'photosynthesis' came into common usage as the term of choice. Later discovery of anoxygenic photosynthetic bacteria and photophosphorylation necessitated redefinition of the term. This essay examines the history of changes in the meaning of photosynthesis. PMID- 16245099 TI - A list of personal perspectives with selected quotations, along with lists of tributes, historical notes, Nobel and Kettering awards related to photosynthesis. AB - The history of photosynthesis research can be found in original papers and books. However, a special history is available from the prefatory chapters and the personal perspectives of various researchers who published them in several journals over the last 40 years. We have compiled a list of such perspectives published since 1964. Selection is not easy, especially of authors who were not directly engaged in photosynthesis research; some are included for their special insights related to central issues in the study of photosynthesis. Our journal, Photosynthesis Research, contains other valuable historic data in the occasional tributes, obituaries and historical notes, that have been published. Lists of these items are included. This article ends by listing the Nobel prizes related to photosynthesis and the Kettering Awards for Excellence in Photosynthesis Research. Wherever possible, a web page address is provided. The web page addresses have been taken from the article 'Photosynthesis and the Web: 2001' by Larry Orr and Govindjee, available at http://www.life.uiuc.edu/govindjee/photoweb and at http://photoscience.la.asu.edu/photosyn/ photoweb/default.html.When I find a bit of leisureI trifle with my papers.This is one of the lesserfrailities.'- Horace, Satires I, IV. PMID- 16245100 TI - In one era and out the other. AB - A guided tour through much of photosynthesis research as I saw it, 1936-2001, is presented here. For earlier perspectives, see Myers 1974 (Plant Physiol 54: 420 426) and 1996 (Photosynth Res 50: 195-208). PMID- 16245101 TI - Following the path of carbon in photosynthesis: a personal story. AB - Chronological recognition of the intermediates and mechanisms involved in photosynthetic carbon dioxide fixation is delineated. Sam Ruben and Martin Kamen's development of application of radioactive carbon for the study of carbon dioxide fixation provided impetus and techniques for following the path of carbon in photosynthesis. Discovery The identity of the primary carboxylation enzyme and its identity with the major protein of photosynthetic tissues ('Fraction 1' protein of Sam Wildman) is reviewed. Memories are dimmed by sixty years of exciting discoveries exploration in newer fields [see Benson 2002 (Annu Rev Plant Biol 53: 1-25), for research and perspectives beyond the early Berkeley days]. PMID- 16245102 TI - ;And whose bright presence' - an appreciation of Robert Hill and his reaction. AB - The Hill reaction, its elucidation, and significance is briefly described. Hill oxidants, the role of the methemoglobin reducing factor and its relation to ferredoxin, and the part played by chloroplast envelopes are discussed. PMID- 16245103 TI - The early history of the genetics of photosynthetic bacteria: a personal account. AB - The development of genetics as a tool for the study of photosynthesis is recounted, beginning in the period when no genetic exchange mechanism was known for any photosynthetic microorganism, and ending with the sequencing of the key genes for photosynthesis. PMID- 16245104 TI - Prochlorophyta - a matter of class distinctions. AB - Prochloron (a marine symbiont) and Prochlorothrix (from freshwater plankton) contain chlorophylls a and b; Prochlorococcus (common in marine picoplankton) contains divinyl-chlorophylls a and b. Like cyanophytes they are all clearly photosynthetic prokaryotes, but since they contain no blue or red bilin pigment they were assigned to a new algal sub-class, the Prochlorophyta. However, since their possible phylogenetic relationships to ancestral green-plant chloroplasts have not received support from molecular biology, it now seems expedient to consider them as aberrant cyanophytes. PMID- 16245105 TI - Research on photosynthetic reaction centers from 1932 to 1987. AB - The history of research on photosynthetic reaction centers is outlined, starting with the implication of their existence through the discovery of the photosynthetic unit, as reported by R. Emerson and W. Arnold in 1932, and culminating in the crystallization and X-ray analysis of the anoxygenic bacterial reaction centers, reported by J. Deisenhofer, H. Michel, and coworkers, over the period 1982-1987. Reaction centers of purple photosynthetic bacteria have received the most attention because they have been well purified and characterized. Structures of cyanobacterial reaction centers of Photosystems I and II are now available from the laboratories of H. Witt and W. Saenger. PMID- 16245106 TI - Excitation energy trapping in anoxygenic photosynthetic bacteria. AB - Various aspects of excitation energy conversion in anoxygenic photosynthetic bacteria are surveyed. This minireview discusses different models that have been proposed during the past 60 years to describe excitation energy transfer from an antenna molecule to the reaction center. First, a simple one-dimensional model was suggested, but over time the models became more detailed when structural and dynamic information was included. This review focuses mainly on the picture of purple bacteria and green sulfur bacteria developed during the past decades. PMID- 16245107 TI - The two-electron gate in photosynthetic bacteria. AB - This paper gives a historical and personal account of the author's work in Rod Clayton's laboratory, when he observed the first evidence of the two-electron gate in bacterial reaction center. Colin Wraight had independently discovered this phenomenon at the same time. The high similarity between the acceptor side of Photosystem II (PS II) and of bacterial reaction centers was one of the first proofs for a profound homology between these two photosystems. PMID- 16245108 TI - Membrane biogenesis in anoxygenic photosynthetic prokaryotes. AB - Following the discovery of photosynthetic bacteria in the nineteenth century, technical developments of the 1950s led to their use in membrane biogenesis studies. These investigations had their origins in the isolation of subcellular particles designated as 'chromatophores' by Roger Stanier and colleagues, which were shown to be photosynthetically competent by Albert Frenkel, and to originate from the intracytoplasmic membrane (ICM) continuum observed in electron micrographs. These ultrastrucutral studies by the G. Drews group, Germaine Cohen Bazire and others also suggested that the ICM originates by invagination of the cytoplasmic membrane, as later established in the biochemical and biophysical work of the R. Niederman and Drews groups. Through a combination of genetic approaches, first introduced in the early 1980s by Barry Marrs, and the atomic resolution structures determined for light-harvesting antennae and reaction centers, a detailed understanding is emerging of mechanisms regulating their levels in the membrane and the roles played by specific protein domains and additional factors in their assembly and supramolecular organization. Prospects for additional progress during the twenty-first century include further elucidation of molecular aspects of the assembly process and the application of newer spectroscopic probes to photosynthetic unit formation. PMID- 16245110 TI - On the natural selection and evolution of the aerobic phototrophic bacteria. AB - This contribution gives a brief survey of the short history since the discovery of the aerobic phototrophic bacteria to focus on a general evolutionary scenario. Most of the citations are of reviews that have covered the earlier literature and to which the reader is directed at appropriate places in the following text. The data summarized in these reviews are supplemented with information from recent or otherwise key primary publications in order to support a synthesis that addresses vexing questions about bacteria containing photosynthetic pigment-protein complexes, but which are incapable of growth with light as the sole, or even the major source of energy. PMID- 16245111 TI - Nitrogen fixation by photosynthetic bacteria. AB - In 1949, Howard Gest and Martin Kamen published two brief papers in Science that changed our perceptions about the metabolic capabilities of photosynthetic bacteria. Their discovery of photoproduction of hydrogen and the ability of Rhodospirillum rubrum to fix nitrogen led to a greater understanding of both processes. PMID- 16245109 TI - Photosynthesis genes and their expression in Rhodobacter sphaeroides 2.4.1: a tribute to my students and associates. AB - This minireview traces the photosynthesis genes, their structure, function and expression in Rhodobacter sphaeroides 2.4.1, as applied to our understanding of the inducible photosynthetic intracytoplasmic membrane system or ICM. This focus has represented the research interests of this laboratory from the late 1960s to the present. This opportunity has been used to highlight the contributions of students and postdoctorals to this research effort. The work described here took place in a much greater and much broader context than what can be conveyed here. The 'timeline' begins with a clear acknowledgment of the work of June Lascelles and William Sistrom, whose foresight intuitively recognized the necessity of a 'genetic' approach to the study of photosynthesis in R. sphaeroides. The 'timeline' concludes with the completed genome sequence of R. sphaeroides 2.4.1. However, it is hoped the reader will recognize this event as not just a new beginning, but also as another hallmark describing this continuum. PMID- 16245112 TI - Photosynthetic exciton theory in the 1960s. AB - Theoretical developments in the 1960s concerning the migration of chlorophyll electronic excitation energy through a photosynthetic core antenna to a reaction center are reviewed in three parts. These include the first theory paper whose calculated results were consistent with experiment, the first analytic determination of the mean number of steps in the two-dimensional random walk of a dephased exciton to reach the reaction center, and the first theoretical description of the possible effects of true collective excited states (Frenkel excitons) on the rate of trap-limited migration and trapping. The possible relevance of these developments, particularly the last, to current photosynthesis research is briefly discussed. PMID- 16245113 TI - Fluorescence lifetime, yield, energy transfer and spectrum in photosynthesis, 1950-1960. AB - The fluorescence lifetime of chlorophyll agives information about the primary photo-physical events in photosynthesis. Most of the light energy absorbed by chlorophylls is utilized for photochemistry. There are two main additional pathways competing for the absorbed light energy: fluorescence and radiationless internal conversion (heat). Only a few percent of the absorbed energy proceeds along these two pathways. This historical minireview focuses on the first direct measurements of the lifetime of chlorophyll fluorescence, the time it takes to transfer energy from phycoerythrin to chlorophyll a, and the discovery of the fluorescence band at 720 nm (F720; then attributed to a dimer of chlorophyll). These works were carried out during the the late 1950s to the early 1960s in the laboratory of Professor Eugene Rabinowitch at the University of Illinois, Urbana Champaign [Brody (1995) Photosynth Res 43: 67-74]. PMID- 16245114 TI - Visualization of excitation energy transfer processes in plants and algae. AB - Development of the time-resolved fluorescence spectroscopy in the pico-second time range and its application to the energy transfer processes in many photosynthetic organisms is reviewed here. This method enabled visualization of energy transfer processes by three-dimensional expression of fluorescence spectra and discrimination of kinetic components and spectral components. The second generation of the ultrafast fluorescence spectroscopy is the femto-second (fs) fluorescence up-conversion, and this has enabled analyses of the transfer processes from carotenoids to chlorophylls with a resolution of less than 100 fs. For future progress, a further development of the spectroscopy is indispensable as well as structural data at atomic resolution. PMID- 16245115 TI - Plastoquinone redox control of chloroplast thylakoid protein phosphorylation and distribution of excitation energy between photosystems: discovery, background, implications. AB - Chloroplast thylakoid protein phosphorylation was discovered, and the most conspicuous phosphoproteins identified, by John Bennett at Warwick University. His initial findings were published in 1977. The phosphoproteins included apoproteins of chloroplast light harvesting complex II. Thylakoid protein phosphorylation was shown to influence distribution of excitation energy between Photosystems I and II in 1979, during a visit by Bennett to the laboratory of Charles J. Arntzen at the University of Illinois at Urbana-Champaign. That work was published by Bennett, Katherine E. Steinback and Arntzen in 1980. Control of both protein phosphorylation and excitation energy distribution by the redox state of the plastoquinone pool was first established in 1980 during the author's visit to Arntzen's laboratory. The experiments were prompted by the realization that coupling between redox state of an inter-photosystem electron carrier and excitation energy distribution provides a concrete mechanism for adaptations known as state transitions. This work was published by Allen, Bennett, Steinback, and Arntzen in 1981. This discovery and its background are discussed, together with some implications for photosynthesis and for research generally. This minireview is a personal account of the Urbana-Warwick and related collaborations in 1979-83: it includes impressions, conjectures, and acknowledgements for which the author is solely responsible. PMID- 16245116 TI - The chequered history of the development and use of simultaneous equations for the accurate determination of chlorophylls a and b. AB - Over the last half century, the most frequently used assay for chlorophylls in higher plants and green algae, the Arnon assay [Arnon DI (1949) Plant Physiol 24: 1-15], employed simultaneous equations for determining the concentrations of chlorophylls a and b in aqueous 80% acetone extracts of chlorophyllous plant and algal materials. These equations, however, were developed using extinction coefficients for chlorophylls a and b derived from early inaccurate spectrophotometric data. Thus, Arnon's equations give inaccurate chlorophyll a and b determinations and, therefore, inaccurate chlorophyll a/b ratios, which are always low. This paper describes how the ratios are increasingly and alarmingly low as the proportion of chlorophyll a increases. Accurate extinction coefficients for chlorophylls a and b, and the more reliable simultaneous equations derived from them, have been published subsequently by many research groups; these new post-Arnon equations, however, have been ignored by many researchers. This Minireview records the history of the development of accurate simultaneous equations and some difficulties and anomalies arising from the retention of Arnon's seriously flawed equations. PMID- 16245117 TI - Changing concepts about the distribution of Photosystems I and II between grana appressed and stroma-exposed thylakoid membranes. AB - Thylakoid membranes of higher plants and some green algae, which house the light harvesting and energy transducing functions of the chloroplast, are structurally unique. The concept of the photosynthetic unit of the 1930s (Robert Emerson, William Arnold and Hans Gaffron), needing one reaction center per hundreds of antenna molecules, was modified by the discovery of the Enhancement effect in oxygen evolution in two different wavelengths of light (Robert Emerson and his coworkers) in the late 1950s, followed by the 1960 Z scheme of Robin Hill and Fay Bendall. It was realized that two light reactions and two pigment systems were needed for oxygenic photosynthesis. Changing ideas about the distribution of Photosystem II (PS II) and PS I between the green-appressed and stroma-exposed thylakoid membrane domains, which led to the concept of lateral heterogeneity, are discussed. PMID- 16245118 TI - Period four oscillations in chlorophyll a fluorescence. AB - The discovery of period four oscillations of the fluorescence yield under flashing light demonstrated that not only the redox state of the Photosystem II (PS II) electron acceptor Q(A), but also the oxygen evolving cycle (described by the S states) modulates the fluorescence yield of chlorophyll (Chl). The positive charges accumulated on the donor side of PS II act on the fluorescence yield (measured in the Q(A) (-) state during a strong flash) through the concentration of the quencher P(680) (+), the oxidized form of PS II reaction center Chl a. However, the period four oscillations of the fluorescence yield detected 1 s after a strong flash (in the P(680)Q(A) state) have not yet been fully explained. PMID- 16245119 TI - Chloride and calcium in Photosystem II: from effects to enigma(*). AB - This minireview focuses on the early evidence for roles of chloride and calcium ions in reactions of photosynthetic electron transport and on the reluctance with which an essential function of these inorganic ions in the process of water oxidation was accepted. For example, Daniel Arnon's group initially refuted the conclusion of Otto Warburg, the discoverer of a 'chloride effect,' that chloride was a 'coenzyme.' Their reasoning was that chloride had not been shown to be an essential mineral nutrient of plants. In the case of calcium, the problem was that the first 'calcium effects' had been seen with preparations from cyanobacteria rather than from green plants. While today the status of calcium and chloride as essential participants in the process of water oxidation is rarely disputed, the nature of their involvement still eludes all experimental inquiries. Substantial progress in this respect may come from recent refinements of the application of Fourier-transform infrared spectroscopy to the study of photosynthetic water oxidation. PMID- 16245120 TI - The bicarbonate effect, oxygen evolution, and the shadow of Otto Warburg. AB - A short list of the twentieth century's dominant figures in photosynthesis would unquestionably include Otto Warburg. One of his many discoveries, the 'bicarbonate effect' remains a lasting puzzle to his heirs in the field. Recent developments in this area of research have renewed interest and call for a re examination of the ideas surrounding this controversial topic. Focus here will be on hypotheses developed by a small number of researchers who proposed that bicarbonate may be involved in oxygen evolution. The effect of bicarbonate on the acceptor side of Photosystem II (PS II) is discussed by Jack van Rensen (in this issue). PMID- 16245121 TI - Role of bicarbonate at the acceptor side of Photosystem II. AB - Besides being the substrate for the carboxylation reaction of photosynthesis, CO(2) (bicarbonate) is required for the activity of Photosystem II (water plastoquinone oxido-reductase). It plays a role on the electron donor side as well as the electron acceptor side. In this contribution, attention will mostly be focused on the history of research into the effects of bicarbonate on electron flow reactions on the acceptor side. Donor side reactions are discussed in this issue by Alan Stemler. PMID- 16245123 TI - P430: a retrospective, 1971-2001. AB - The spectral species P430 and its spectral and kinetic properties are briefly reviewed. Currently available evidence shows P430 to be the optic-spectral representation of FeS-A/B, the electron acceptor(s) of Photosystem I (PS I). PMID- 16245122 TI - Photosystem I reaction center: past and future. AB - Science has always been drawn to uncover fundamental life processes. Photosynthesis is one, if not the most fascinating, of them. Within it, the protein complexes that catalyze light-induced electron transport and photophosphorylation are enchanting creations of evolution. Plant Photosystem I (PS I) is not the largest protein complex in nature but it is the most elaborate in the number of prosthetic groups involved in its fabric. Thirty years ago, one of us (NN) developed a fascination for this complex and, despite the apparent neglect (lack of publications in the last few years), never let it go. Only a crystal structure at 2 A resolution will satiate our curiosity. In this minireview, we trace the past, and end the article with a comment on future prospects. For the present situation, see Parag Chitnis (2001). PMID- 16245124 TI - The ferredoxin/thioredoxin system: from discovery to molecular structures and beyond. AB - Experiments initiated in the early 1960s on fermentative bacteria led to the discovery of ferredoxin-dependent alpha-ketocarboxylation reactions that were later found to be key to a new cycle for the assimilation of carbon dioxide in photosynthetic bacteria (the reductive carboxylic acid or reverse citric cycle). The latter finding set the stage for the discovery of a regulatory system, the ferredoxin/thioredoxin system, functional in photosynthesis in chloroplasts and oxygen-evolving photosynthetic prokaryotes. The chloroplast research led to a description of the extraplastidic NADP/thioredoxin system that is now known to function in heterotrophic plant processes such as seed germination and self incompatibility. Extensions of the fundamental research have begun to open doors to the broad application of thioredoxin in technology and medicine. PMID- 16245125 TI - Irrungen, Wirrungen? The Mehler reaction in relation to cyclic electron transport in C3 plants. AB - Plants not only evolve but also reduce oxygen in photosynthesis. Considerable oxygen uptake occurs during photorespiration of C3 plants. Controversies exist on whether direct oxygen reduction in the Mehler reaction together with associated electron transport is also a major sink of electrons when leaves are exposed to sunlight. Here, preference is given to the view that it is not. Whereas photorespiration consumes ATP, the Mehler reaction does not. In isolated chloroplasts photosynthesizing in the presence of saturating bicarbonate, the Mehler reaction is suppressed. In the water - water cycle of leaves, which includes the Mehler reaction, water is oxidized and electrons flow through Photosystems II and I to oxygen producing water. The known properties of coupled electron transport suggest that the water - water cycle cannot act as an efficient electron sink. Rather, by contributing to thylakoid acidification it plays a role in the control of Photosystem II activity. Cyclic electron transport competes with the Mehler reaction for electrons. Both pathways can help to defray possible ATP deficiencies in the chloroplast stroma, but play a more important role by making intrathylakoid protein protonation possible. This is a necessary step for the dissipation of excess excitation energy as heat. Linear electron flow to oxygen relieves the inhibition of cyclic electron transport, which is observed under excessive reduction of intersystem electron carriers. In turn, cyclic electron transport replaces functions of the linear pathway in the control of Photosystem II when oxygen reduction is decreased at low temperatures or, experimentally, when the oxygen concentration of the gas phase is low. Thus, cyclic electron flow acts in flexible relationship with the water-water cycle to control Photosystem II activity. PMID- 16245127 TI - Along the trail from Fraction I protein to Rubisco (ribulose bisphosphate carboxylase-oxygenase). AB - This historical minireview deals with events leading to the eventual discovery of Rubisco (ribulose bisphosphate carboxylase-oxygenase). This abundant leaf protein is not only responsible for the net fixation of CO(2) in all plants, but also causes the loss of carbon through photorespiration. The latter is a special 'problem' of the so-called 'C(3)' plants. The protein was first called 'Fraction 1 protein' before it was recognized to be the same as Rubisco. Instead of reinventing words, text as needed has been freely used from three earlier publications (Wildman and Kwanyuen 1978; Wildman 1992, 1998). PMID- 16245126 TI - Photophosphorylation and the chemiosmotic perspective. AB - Photophosphorylation was discovered in chloroplasts by D. Arnon and coworkers, and in bacterial 'chromatophores' (intercytoplasmic membranes) by A. Frenkel. Initial low rates were amplified by adding electron-carrying compounds such as FMN, later shown to support the 'pseudocyclic' electron flow. ATP synthesis, and coupling to electron flow, was detected accompanying linear electron flow from H(2)O to either NADP(+) or ferricyanide. Another pattern of electron flow supporting photophosphorylation was that of a cycle around Photosystem I (PS I). Isolation and analysis of the ATP synthase showed, as with mitochondrial and bacterial analogues, an intrinsic membrane complex (CF(0)) and an extrinsic complex (CF(1)). CF(1) is a latent ATPase, activated additively by the high energy state of the thylakoids, and by reduction of a disulfide bond on the gamma subunit. Once reduced, ATP synthesis occurs at lower energy levels. The search for an 'intermediate' linking electron flow and ATP synthesis led to the discovery of post-illumination ATP synthesis by thylakoids, where turnover occurs in the dark. Once interpreted by P.Mitchell's chemiosmotic hypothesis, this led to the discovery of light-driven proton uptake into the thylakoid lumen, with accompanying Cl(-) intake and Mg(2+) and K(+) output. Chemiosmosis was confirmed in several ways, including ATP synthesis in the dark due to an acid-to-base transition of thylakoids, and photophosphorylation accomplished in artificial lipid vesicles containing both the proton-pumping bacterial rhodopsin and a mitochondrial ATPase complex. The now generally accepted chemiosmotic interpretation is able to clarify some other aspects of photosynthesis as well. PMID- 16245128 TI - C(4) photosynthesis: discovery and resolution. AB - This Minireview provides a brief account of the scene and interesting turn of events surrounding the discovery and resolution of the mechanism of C(4) photosynthesis, as well as the recognition of the process by the wider plant science community. PMID- 16245129 TI - The discovery of Rubisco activase - yet another story of serendipity. AB - A brief history of Rubisco (ribulose bisphosphate carboxylase oxygenase) research and the events leading to the discovery and initial characterization of Rubisco activase are described. Key to the discovery was the chance isolation of a novel Arabidopsis photosynthesis mutant. The characteristics of the mutant suggested that activation of Rubisco was not a spontaneous process in vivo, but involved a heritable factor. The search for the putative factor by 2D electrophoresis identified two polypeptides, genetically linked to Rubisco activation, that were missing in chloroplasts from the mutant. An assay for the activity of these polypeptides, which were given the name Rubisco activase, was developed after realizing the importance of including ribulose bisphosphate (RuBP) in the assay. The requirement for ATP and the subsequent identification of activase as an ATPase came about fortuitously, the result of a RuBP preparation that was contaminated with adenine nucleotides. Finally, the ability of activase to relieve inhibition of the endogenous Rubisco inhibitor, 2-carboxyarabinitol 1 phosphate, provided an early indication of the mechanism by which activase regulates Rubisco. PMID- 16245130 TI - Three decades in transport business: studies of metabolite transport in chloroplasts - a personal perspective. AB - This article gives an historical overview of our group's research on various metabolite translocators of chloroplasts, such as the translocators for phosphorylated intermediates of the Calvin-Benson cycle and of glycolysis, of ADP and ATP, of dicarboxylates, of pyruvate and of hexoses; how it began and where it led to. Wherever appropriate, references will be made to research in other laboratories. PMID- 16245131 TI - The present model for chlororespiration. AB - The present model of chlororespiration deals with the dark reduction and oxidation of plastoquinone. Both stages are reviewed here for algae and higher plants. Recent data confirm the presence of a plastoquinone:oxygen oxidoreductase with features different from those of the mitochondrial oxidases. The possible involvement of the chloroplast oxidase in the pathway of carotenoid biosynthesis is discussed in view of various experimental data and on energetics considerations. PMID- 16245132 TI - Gene-targeted and site-directed mutagenesis of photosynthesis genes in cyanobacteria. AB - This historical minireview traces the development and application of methods for gene-targeted and site-directed mutagenesis of photosynthesis genes in cyanobacteria (mainly Synechocystis sp. PCC 6803). This approach allowed important data to be obtained on the structure and function of Photosystem I and Photosystem II complexes. I describe some of the major contributions of molecular genetics and subsequent mutant analysis in the 1980s and early 1990s that led to substantial advances in our knowledge of basic principles regarding the organization of the photosynthetic apparatus. This molecular-genetic research on cyanobacteria has initiated a fresh wave of photosynthesis research and created a solid foundation for rapid progress at the threshold of the twenty-first century. PMID- 16245133 TI - The three genomes of Chlamydomonas. AB - During the past 50 years, the green unicellular alga Chlamydomonas reinhardtii has played a key role as model system for the study of photosynthesis and chloroplast biogenesis. This is due to its well-established nuclear and chloroplast genetics, its dispensable photosynthetic function in the presence of acetate, and its highly efficient nuclear and chloroplast transformation systems. Considerable progress has been achieved in our understanding of the structure, function, inheritance, and expression of nuclear, chloroplast, and mitochondrial genes and of the molecular cross-talk between the nuclear, chloroplast, and mitochondrial genetic systems. PMID- 16245134 TI - The Laboratory of Photosynthesis and its successors at Gif-sur-Yvette, France. AB - This article is not a survey of all the research made during the last half century at the 'Laboratoire de Photosynthese' of the 'Centre National de la Recherche Scientifique' (CNRS) in Gif-sur-Yvette, but rather some personal recollections, as faithful as possible. Not all people could be mentioned and the scientists named here are mainly those who, at different stages of the laboratory's evolution, created their research teams, within or outside the laboratory. The laboratory, closed now as an administrative entity, was founded in 1953 by the CNRS in Gif-sur-Yvette, near Paris. Besides the emerging research groups in Paris and at Saclay, it was then the only one in France to be entirely dedicated to photosynthesis. Initially, the focus was on metabolic biochemistry of photosynthesis in whole plants and unicellular algae. In 1959, biophysics of primary and associated processes was added and in 1966, the laboratory was enlarged to include molecular genetics and, somewhat later, structural biology. Most of the early members of the laboratory have now gone offstage, but the research goes on, in Gif and elsewhere, thanks to the numerous high-level scientists that have been trained there. Most of the basic questions have now been answered, and interest has shifted in two directions, atomic and integrated, while many other facets of research are no longer specific to photosynthesis but part of more general biological problems, a normal situation for an area that has reached its maturity. PMID- 16245136 TI - Contributions of the free oxidized and Q(B)-bound plastoquinone molecules to the thermal phase of chlorophyll-a fluorescence. AB - Variable chlorophyll a (Chl a) fluorescence is composed of a photochemical and a thermal phases of similar amplitudes. The photochemical phase can be induced by a saturating single turnover flash (STF) and reflects the reduction of the Photosystem II (PS II) Q(A) primary electron acceptor. The thermal phase requires multiple turnover flash (MTF) and is somehow related to the reduction of the plastoquinone (PQ) molecules. This article aimed to determine the relative contributions of the Q(B)-bound and the free oxidized PQ molecules to the thermal phase of Chl a fluorescence. We thus measured the interactive effects of exogenous PQ (PQex), of an inhibitor (DCMU) acting at the Q(B) site of PS II and of an artificial quencher, 2-methyl-1,4-naphtoquinone, on Chl a fluorescence levels induced by STF (F(F)) and MTF (F(M)) in spinach thylakoids. We observed that: (1) the incorporation of PQex in thylakoids stimulated photosynthetic electron transport but barely affected F(F) and F(M) in the absence of DCMU; (2) DCMU significantly increased the amplitude of F(F) but slightly quenched F(M); (3) 2-methyl-1,4-naphtoquinone quenched F(M) to a larger-extent than F(F); (4) DCMU increased the quenching effects of PQex on F(F) and F(M) and also, of methyl 1,4-naphtoquinone on F(F). These results indicate that: (1) the Q(B)-bound and the free PQ molecules contribute to about 56% and 25%, respectively, to the thermal phase Chl a fluorescence in dark-adapted thylakoids; and (2) the thermal phase of Chl a fluorescence is more susceptible than the photochemical phase to the non-photochemical quenching effect of oxidized quinones. PMID- 16245135 TI - Chloroplast fructose-1,6-bisphosphatase: structure and function. AB - Redox regulation of photosynthetic enzymes has been a preferred research topic in recent years. In this area chloroplast fructose-1,6-bisphosphatase is probably the most extensively studied target enzyme of the CO(2) assimilation pathway. This review analyzes the structure, biosynthesis, phylogeny, action mechanism, regulation and kinetics of fructose-1,6-bisphosphatase in the light of recent findings on structure-function relationship, and from a molecular biology viewpoint. PMID- 16245137 TI - Plant peroxiredoxins: alternative hydroperoxide scavenging enzymes. AB - The role of plant peroxiredoxins in the detoxification systems is discussed in relation with the existence of many isoforms of this protein in distinct plant compartments. Phylogenetic analyses indicate that plant peroxiredoxins can be divided into four classes. Two of these classes correspond to chloroplastic enzymes. All isoforms contain at least one conserved catalytic cysteine. The enzymes belonging to the 1-Cys Prx class seem to be seed restricted and to play a role of detoxification during the germination process. At least one putative cytosolic isoform can use both thioredoxin and glutaredoxin as an electron donor, but the chloroplastic isoforms characterized depend on reduced thioredoxin. Mutagenesis and plant transformation studies support the proposal that the chloroplastic peroxiredoxins play an important role in combating the ROS species generated at the level of the chloroplastic electron transfer chain. PMID- 16245138 TI - Transcription of three sets of genes coding for the core light-harvesting proteins in the purple sulfur bacterium, Allochromatium vinosum. AB - The nucleotide sequence of the puf operon coding for the subunits of the photosynthetic reaction center and the core light-harvesting complex (LH1) of the purple sulfur bacterium, Allochromatium (A.) vinosum (formally Chromatium vinosum), was completely determined. Unlike other known puf operons, which contain only one set of genes coding for the LH1 apoproteins, pufB and pufA, the A. vinosum puf operon included three sets of pufB and pufA genes with a gene order of pufB (1) A (1) LMCB (2) A (2) B (3) A (3). Northern hybridization analysis suggested that all of the nine puf genes are co-transcribed as a 4.43 kb mRNA. Three small mRNAs corresponding to pufB (2) A (2) B (3) A (3), pufB (2) A (2) B (3), and pufB (2) A (2) were detected, as well as two small mRNAs covering pufB (1) A (1). Analysis of the nucleotide sequence of the puf operon, including the flanking regions and 5'-ends of the six mRNAs, suggested that the transcription of the A. vinosum puf operon is initiated at 74 bp downstream from the bchZstop codon (295 bp upstream from the pufB (1) start codon), and regulated by a promoter located at its direct upstream. The possible promoter is overlapped with a binding motif of a repressor protein for pigment-biosynthesis genes, PpsR or CrtJ, known in other purple bacteria. No other possible promoters were found within the puf genes. These findings indicate that three sets of pufA and pufB genes of A. vinosum are co-transcribed as a long mRNA containing all the puf genes, and, from this long mRNA, the five short mRNAs are possibly derived by post-transcriptional modifications. PMID- 16245139 TI - An in situ study of photosynthetic oxygen exchange and electron transport rate in the marine macroalga Ulva lactuca (Chlorophyta). AB - Direct comparisons between photosynthetic O(2) evolution rate and electron transport rate (ETR) were made in situ over 24 h using the benthic macroalga Ulva lactuca (Chlorophyta), growing and measured at a depth of 1.8 m, where the midday irradiance rose to 400-600 mumol photons m(-2) s(-1). O(2) exchange was measured with a 5-chamber data-logging apparatus and ETR with a submersible pulse amplitude modulated (PAM) fluorometer (Diving-PAM). Steady-state quantum yield ((F(m)'-F(t))/F(m)') decreased from 0.7 during the morning to 0.45 at midday, followed by some recovery in the late afternoon. At low to medium irradiances (0 300 mumol photons m(-2) s(-1)), there was a significant correlation between O(2) evolution and ETR, but at higher irradiances, ETR continued to increase steadily, while O(2) evolution tended towards an asymptote. However at high irradiance levels (600-1200 mumol photons m(-2) s(-1)) ETR was significantly lowered. Two methods of measuring ETR, based on either diel ambient light levels and fluorescence yields or rapid light curves, gave similar results at low to moderate irradiance levels. Nutrient enrichment (increases in [NO(3) (-)], [NH(4) (+)] and [HPO(4) (2-)] of 5- to 15-fold over ambient concentrations) resulted in an increase, within hours, in photosynthetic rates measured by both ETR and O(2) evolution techniques. At low irradiances, approximately 6.5 to 8.2 electrons passed through PS II during the evolution of one molecule of O(2), i.e., up to twice the theoretical minimum number of four. However, in nutrient-enriched treatments this ratio dropped to 5.1. The results indicate that PAM fluorescence can be used as a good indication of the photosynthetic rate only at low to medium irradiances. PMID- 16245140 TI - Characterization of P700 as a photochemical quencher in isolated Photosystem I particles using simultaneous measurements of absorbance changes at 830 nm and photoacoustic signal. AB - The relationship between the redox state of P700, the primary donor of PS I, monitored using absorbance changes at 830 nm and photochemical energy storage in PS I reaction centers assayed with the photoacoustic method (PA) was studied in isolated PS I submembrane particles aspirated onto nitrocellulose filters. Several donors have been used to support the electron transport through PS I. NADPH and NADH demonstrated low rates of electron donation to PS I, while ascorbate and ascorbate plus 2,6-dichlorophenolindophenol (DCIP) couple have been found more effective in both P700(+) reduction and stimulation of the variable component of the PA signal. A linear relationship was found in isolated PS I particles between the (A(830,max) - A(830,steady))/A(830,max) and (PA(max) - PA(steady))/PA(max) ratios, which characterized the relative amount of P700 in the reduced state and the relative magnitude of the variable PA component, respectively. That linear relationship was obtained independently from the nature of electron donor used for the reduction of P700(+). Such linear relationship was also obtained at various wavelengths of modulated light in the range of 660 to 720 nm, only the slope of the linear fits varied with wavelength. It is concluded that reduced P700 act as a photochemical quencher of absorbed energy. Variable thermal dissipation in PS I reaction centers of isolated submembrane particles linearly depends on the amount of reduced P700 and thus constitutes an appropriate indicator of the redox pressure applied to PS I. PMID- 16245141 TI - Non-photochemical quenching of chlorophyll fluorescence in Chlorella fusca acclimated to constant and dynamic light conditions. AB - Non-photochemical quenching of chlorophyll fluorescence (NPQ) involves dissipation of light energy in the photosynthetic apparatus via a number of physiologically distinct processes. The relationships among NPQ, the (de)epoxidation state of the xanthophyll cycle pigments and state transitions was studied in the green alga Chlorella fusca, acquired from six differently light acclimated continuous cultures. A 10 h light and 14 h darkness, periodicity was obeyed in all cultures. Three cultures received a high total daily irradiance, three others a low one. High and low irradiances were each dosed in three different modes at constant supply, with sine shape intensity modulation, or as a sine with superimposed oscillations. In the constant supply mode, but not for the sine and oscillating modes, high-light rendered a three-fold higher xantophyll cycle pigment content than low-light. Dynamic interconversion of xantophyll cycle pigments was restricted to high-light cultures. NPQ followed the kinetics of the light supply mode and was highest in high light cultures. In low-light cultures, NPQ correlated mainly to state transitions. These observations were supported by experiments with dithiothreithol-treated samples. The relative impact of xantophyll cycle operation and state transitions on NPQ in green algae from different light climates will be discussed with reference to higher plants. PMID- 16245142 TI - New type of dual-channel PAM chlorophyll fluorometer for highly sensitive water toxicity biotests. AB - A new type of dual-channel PAM chlorophyll fluorometer has been developed, which is specialised in the detection of extremely small differences in photosynthetic activity in algae or thylakoids suspensions. In conjunction with standardised algae cultures or isolated thylakoids, the new device provides an ultrasensitive biotest system for detection of toxic substances in water samples. In this report, major features of the new device are outlined and examples of its performance are presented using suspensions of Phaeodactylum tricornutum (diatoms) and of freeze-dried thylakoids of Lactuca sativa (salad). Investigated and reference samples are exposed to the same actinic intensity of pulse modulated measuring light. The quantum yields are assessed by the saturation pulse method. Clock-triggered repetitive measurements of quantum yield typically display a standard deviation of 0.1%, corresponding to the inhibition induced by 0.02 mug diuron l(-1). Hence, for diuron or compounds with similar toxicity, the detection limit is well below the 0.1 mug l(-1) defined as the limit for the presence of a single toxic substance in water by the European Commission drinking water regulation. The amounts of water and biotest material required for analysis are very small, as a single assay involves two 1 ml samples, each containing ca. 0.5 mug chlorophyll. Both with Phaeodactylum and thylakoids the relationship between inhibition and diuron concentration is strictly linear up to 10% inhibition, with very similar slopes. Apparent inhibition depends on the actinic effect of the measuring light, showing optima at 6 and 4 mumol quanta m(-2) s(-1) with Phaeodactylum and thylakoids, respectively. PMID- 16245143 TI - Photosynthesis and phage: early studies on phosphorus metabolism in photosynthetic microorganisms with (32)P, and how they led to the serendipic discovery of (32)P-decay suicide of bacteriophage. AB - During my PhD thesis research (1946-1949), I explored the effects of light on the uptake of (32)P-labeled inorganic phosphate (P(i)) by cells of photosynthetic bacteria and microalgae, and the dynamics of P turnover between low and high molecular weight cell constituents. The results were interpreted as evidence for the conversion of light energy to the chemical energy of phosphorylated compounds. The experimental results also suggested to me that the precursors of the P in DNA bacteriophages of Escherichia coli must be low molecular weight phosphorylated compounds present within the host cells and led to the design of an experiment to determine the conservation of (32)P of an infecting phage particle in its numerous progeny. The experiment envisaged was never conducted because phage labeled with (32)P of high specific activity showed unexpected loss of viability. Thus, by serendipity, 'suicide' of phage due to (32)P-beta decay was discovered. (32)P-decay 'suicide' provided a technique that was useful for analysis of phage genetic structure and replication. This memoir describes the unusual circumstances leading to the decisive role of serendipity in revealing an extraordinary phenomenon. PMID- 16245145 TI - Meiotic transverse filament proteins: essential for crossing over. AB - Meiosis is a specialized set of two nuclear divisions, meiosis I and II, by which a diploid cell produces four haploid daughters. After premeiotic DNA replication, homologous chromosomes pair and recombine, and then disjoin at meiosis I. Subsequently, at meiosis II, the sister chromatids of each chromosome segregate. In nearly all eukaryotes, meiotic chromosome pairing culminates in the formation of a ladder-like supramolecular protein structure, the synaptonemal complex (SC) (Page and Hawley, 2004). The rungs of the ladder are known as transverse filaments (TFs). Genes encoding TF proteins have been identified in a limited number of organisms, and their function has been studied by mutational analysis. Although TF proteins show little amino acid sequence conservation, their structure and function are largely conserved. In all analyzed species, TF proteins are required for meiotic reciprocal recombination (crossing over). PMID- 16245146 TI - Arabidopsis thaliana full genome longmer microarrays: a powerful gene discovery tool for agriculture and forestry. AB - Sequenced plant genomes provide a large reservoir of known genes with potential for use in crop and tree improvement, but assignment of specific functions to annotated genes in sequenced plant genomes remains a challenge. Furthermore, most plant genes belong to families encoding proteins with related but distinct functions. In this commentary, we discuss our development of Arabidopsis spotted whole genome longmer oligonucleotide microarrays, and their use in global transcription profiling. We show that longmer array based transcriptome analysis in Arabidopsis can be used as an efficient and effective gene discovery and functional genomics tool, particularly for functional analyses of members of large gene families. We discuss experiments that focus on gene families involved in phenylpropanoid natural product biosynthesis and fiber differentiation. These analyses have helped to elucidate functions of individual gene family members, and have identified new candidate genes involved in fiber development and differentiation. Results obtained by these studies in Arabidopsis can be used as the basis for gene discovery in commercially important plants, and we have focused our attention on Populus trichocarpa (poplar), a species important in forestry and agroforestry for which complete genome sequence information is available. PMID- 16245147 TI - Engineering disease resistant cattle. AB - Mastitis is a disease of the mammary gland caused by pathogens that find their way into the lumen of the gland through the teat canal. Mammary gland infections cost the US dairy industry approximately $2 billion dollars annually and have a similar impact in Europe. In the absence of effective treatments or breeding strategies to enhance mastitis resistance, we have created transgenic dairy cows that express lysostaphin in their mammary epithelium and secrete the antimicrobial peptide into milk. Staphylococcus aureus, a major mastitis pathogen, is exquisitely sensitive to lysostaphin. The transgenic cattle resist S. aureus mammary gland challenges, and their milk kills the bacteria, in a dose dependent manner. This first step in protecting cattle against mastitis will be followed by introduction of other genes to deal with potential resistance issues and other mastitis causing organisms. Care will be taken to avoid altering milk's nutritional and manufacturing properties. Multi-cistronic constructs may be required to achieve our goals as will other strategies possibly involving RNAi and gene targeting technology. This work demonstrates the possibility of using transgenic technology to address disease problems in agriculturally important species. PMID- 16245144 TI - Transgenic modifications of the rat genome. AB - The laboratory rat (R. norvegicus) is a very important experimental animal in several fields of biomedical research. This review describes the various techniques that have been used to generate transgenic rats: classical DNA microinjection and more recently described techniques such as lentiviral vector mediated DNA transfer into early embryos, sperm-mediated transgenesis, embryo cloning by nuclear transfer and germline mutagenesis. It will also cover techniques associated to transgenesis such as sperm cryopreservation, embryo freezing and determination of zygosity. The availability of several technologies allowing genetic manipulation in the rat coupled to genomic data will allow biomedical research to fully benefit from the rat as an experimental animal. PMID- 16245148 TI - Production of human serum albumin by sugar starvation induced promoter and rice cell culture. AB - Human serum albumin (HSA) is the most widely used clinical serum protein. Currently, commercial HSA can only be obtained from human plasma, due to lack of commercially feasible recombinant protein expression systems. In this study, inducible expression and secretion of HSA by transformed rice suspension cell culture was established. Mature form of HSA was expressed under the control of the sucrose starvation-inducible rice alpha Amy3 promoter, and secretion of HSA into the culture medium was achieved by using the alpha Amy3 signal sequence. High concentrations of HSA were secreted into culture medium in a short time (2-4 days) by sucrose depletion after cell concentrations had reached a peak density in culture medium containing sucrose. The recombinant HSA had the same electrophoretic mobility as commercial HSA and was stable and free from apparent proteolysis in the culture medium. In a flask scale culture with repeated sucrose provision-depletion cycles, HSA was stably produced with yields up to 11.5% of total medium proteins or 15 mg/L per cycle after each sucrose provision-depletion cycle. A bubble column type bioreactor was designed for production of HSA. In the bioreactor scale culture, HSA was produced with yields up to 76.4 mg/L 4 days after sucrose depletion. HSA was purified from the culture medium to high purity by a simple purification scheme. Enrichment of HSA in culture medium simplifies downstream purification, minimizes protease degradation, and may reduce production cost. The combination of a DNA construct containing the alpha Amy3 promoter and signal sequence, and the use of a rice suspension cell culture can provide an effective system for the production of recombinant pharmaceutical proteins. PMID- 16245149 TI - Improvement of human lysozyme expression in transgenic rice grain by combining wheat (Triticum aestivum) puroindoline b and rice (Oryza sativa) Gt1 promoters and signal peptides. AB - Heterologous protein expression levels in transgenic plants are of critical importance in the production of plant-made pharmaceuticals (PMPs). We studied a puroindoline b promoter and signal peptide (Tapur) driving human lysozyme expression in rice endosperm. The results demonstrated that human lysozyme expressed under the control of the Tapur cassette is seed-specific, readily extractable, active, and properly processed. Immuno-electron microscopy indicated that lysozyme expressed from this cassette is localized in protein bodies I and II in rice endosperm cells, demonstrating that this non-storage promoter and signal peptide can be used for targeting human lysozyme to rice protein bodies. We successfully employed a strategy to improve the expression of human lysozyme in transgenic rice grain by combining the Tapur cassette with our well established Gt1 expression system. The results demonstrated that when the two expression cassettes were combined, the expression level of human lysozyme increased from 5.24 +/- 0.34 mg(-1) g flour for the best single cassette line to 9.24 +/- 0.06 mg(-1) g flour in the best double cassette line, indicating an additive effect on expression of human lysozyme in rice grain. PMID- 16245150 TI - Partial correction of abnormal cardiac development in caspase-8-deficient mice by cardiomyocyte expression of p 35. AB - Baculovirus p 35 protein protects cells from apoptotic cell death by inhibiting caspase activation. We have established transgenic mouse lines specifically expressing p 35 in cardiomyocytes, and primary cardiomyocytes isolated from these mice exhibit resistance to staurosporine-induced apoptosis. In a previous study, we observed defects in heart formation associated with abdominal hemorrhage and cardiomyocyte cell death in caspase-8-deficient animals. In order to better understand the etiology of the cardiac defects and embryonic lethality in caspase 8-deficient mice, we crossed these mice with the p 35 transgenic animals. Although the newly generated mice still died in utero and exhibited some cardiac defects, cardiomyocyte apoptosis was suppressed and ventricular trabeculation was restored. Thus, cardiomyocyte expression of p 35 prevented cell death induced by staurosporine or caspase-8 deficiency. Additionally, our data suggest that caspase-8 plays multiple roles in cardiac development. PMID- 16245152 TI - Modification of cytokinin levels in potato via expression of the Petunia hybrida Sho gene. AB - The Sho gene from Petunia hybrida encodes an enzyme for cytokinin synthesis. Here we report on the effects of Shogene expression on potato development. In contrast to transgenic potato expressing the Agrobacterium ipt gene, moderate Sho expression resulted in sufficient root development that allowed the cultivation of the Sho transformants in soil. The most pronounced effects detectable in these lines were an enhanced shoot production, delayed tuber formation, significant reduction in tuber size, and inhibition of tuber dormancy. Sho expression predominantly associated with a strong increase in 2iP glucosides, accompanied by an increase in zeatin glucosides in lines with very high Sho expression levels. The data demonstrate that it is possible to produce viable plants with enhanced cytokinin levels via constitutive Sho expression, which allows an assessment of cytokinin effects in all organs. PMID- 16245151 TI - Inducible excision of selectable marker gene from transgenic plants by the cre/lox site-specific recombination system. AB - In a plant transformation process, it is necessary to use marker genes that allow the selection of regenerated transgenic plants. However, selectable marker genes are generally superfluous once an intact transgenic plant has been established. Furthermore, they may cause regulatory difficulties for approving transgenic crop release and commercialization. We constructed a binary expression vector with the Cre/lox system with a view to eliminating a marker gene from transgenic plants conveniently. In the vector, recombinase gene cre under the control of heat shock promoter and selectable marker gene nptII under the control of CaMV35S promoter were placed between two lox P sites in direct orientation, while the gene of interest was inserted outside of the lox P sites. By using this vector, both cre and nptII genes were eliminated from most of the regenerated plants of primary transformed tobacco through heat shock treatment, while the gene of interest was retained and stably inherited. This auto-excision strategy, mediated by the Cre/lox system and subjected to heat shock treatment to eliminate a selectable marker gene, is easy to adopt and provides a promising approach to generate marker-free transgenic plants. PMID- 16245153 TI - Over-expression of Chinese cabbage calreticulin 1, BrCRT1, enhances shoot and root regeneration, but retards plant growth in transgenic tobacco. AB - Calreticulin (CRT) is a ubiquitously expressed, high capacity Ca(2+)-binding protein that is involved in intracellular Ca(2+) homeostasis and molecular chaperoning in the endoplasmic reticulum (ER). A cDNA encoding a calreticulin, BrCRT1 (Brassica rapa Calreticulin 1), has been isolated from Chinese cabbage (B. rapa subsp. pekinensis) flower bud. Constitutive over-expression of the BrCRT1 gene promotes robust shoot production and root formation at sub-optimal concentrations of BA/NAA, which are important factors controlling plant regeneration in tissue culture. In contrast, the suppressed BrCRT1 line exhibited a slight reduction of shoot and root regeneration. In spite of enhanced regeneration in tissue culture, the seedling and plant growth rate was inhibited in soil. The steady state level of BrCRT1 transcripts was sensitive to exogenous auxins and cytokinins, and rapidly accumulated within 30 min, and this induction required de novo protein synthesis. Together with the results of transgenic tobacco plants and mRNA analysis in Chinese cabbage, our data suggest that BrCRT1 genes may up-regulate the competency of vegetative tissue to respond to hormonal signals involved in shoot and root regeneration processes. PMID- 16245154 TI - Optimization of Acidothermus cellulolyticus endoglucanase (E1) production in transgenic tobacco plants by transcriptional, post-transcription and post translational modification. AB - An attempt was made to obtain a high-level production of intact Acidothermus cellulolyticus endoglucanase (E1) in transgenic tobacco plants. The E1 expression was examined under the control of the constitutive and strong Mac promoter or light-inducible tomato Rubisco small sub-unit (RbcS-3C) promoter with its original or Alfalfa Mosaic Virus (AMV) RNA4 5'-untranslated leader (UTL) and targeted to different sub-cellular compartments via transit peptides. The transit peptides included native E1, endoplasmic reticulum, vacuole, apoplast, and chloroplast. E1 expression and its stability in transgenic plants were determined via E1 activity, protein immunoblotting, and RNA gel-blotting analyses. Effects of sub-cellular compartments on E1 production and its stability were determined in transgenic tobacco plants carrying one of six transgene expression vectors, where the E1 was under the control of Mac promoter, mannopine synthase transcription terminator, and one of the five transit peptides. Transgenic tobacco plants with an apoplastic transit peptide had the highest average E1 activity and protein accumulation, which was about 0.25% of total leaf soluble proteins estimated via E1 specific activity and protein gel blots. Intercellular fluid analyses confirmed that E1 signal peptide functioned properly in tobacco cells to secret E1 protein into the apoplast. By replacing RbcS-3C UTL with AMV RNA4 UTL E1 production was enhanced more than twofold, while it was less effective than the mannopine synthase UTL. It was observed that RbcS-3C promoter was more favorable for E1 expression in transgenic plants than the Mac promoter. E1 activity in dried tobacco seeds stored one year at room temperature was 45% higher than that observed immediately after harvesting, suggesting that E1 protein can be stored at room temperature for a long period. E1 stability in different sub-cellular compartments and the optimal combination of promoter, 5' UTL, and sub-cellular compartmentation for heterologous protein production in transgenic plants are discussed. PMID- 16245155 TI - In vivo Cre/loxP mediated recombination in mouse Clara cells. AB - In small airways, Clara cells are the main epithelial cell type and play an important physiological role in surfactant production, protection against environmental agents, regulation of inflammatory and immune responses in the respiratory system. Thus, Clara cells are involved in lung homeostasis and pathologies like asthma, Chronic Obstructive Pulmonary Diseases (COPD) or cancers. To date, Clara cells implication in these pathological processes remains largely enigmatic. The engineering of a transgenic strain mouse allowing specific gene invalidation in Clara cells may be of interest to improve our knowledge about the genes involved in these diseases. By using the Cre/loxP strategy we report the engineering of a transgenic mouse strain with expression of Cre recombinase under the control of the Clara Cell Secretory Protein (CCSP) promoter. Specific staining and immuno-histochemistry performed after breeding with reporter mice revealed that CCSP drives a functional Cre expression specifically in Clara cells. This mouse strain is a powerful tool for Cre-loxP mediated conditional recombination in the lung and represents a new tool to study Clara cell physiology. PMID- 16245156 TI - Gastric pit cell hyperplasia and glandular atrophy in carbonic anhydrase IX knockout mice: studies on two strains C57/BL6 and BALB/C. AB - Carbonic anhydrase (CA) isoenzyme IX is a hypoxia-inducible enzyme, which is expressed in the human and rodent gastrointestinal tract and overexpressed in several different tumors. Functionally, it has probably an effect on proliferation and differentiation of gastrointestinal epithelial cells. It may also participate in gastric morphogenesis, since a recent study has shown gastric pit cell hyperplasia and glandular atrophy in CA IX-knockout mice. However, it is not known whether CA IX produces morphological changes in the gastric mucosa, which can turn into a dysplasia or malignancy in the presence of some carcinogenic factors. High-salt diet is considered such a factor which has been shown to modulate Helicobacter pylori-associated carcinogenesis. We produced two strains of CA IX-knockout mice, C57/BL6 and BALB/c, and the mice ate either standard or high-salt feed for 20 weeks. Stomach samples were collected from 40 Car 9(-/-) knockout mice and 37 wildtype littermates, and the tissue sections were examined for histology. CA IX-deficiency caused gastric pit cell hyperplasia and glandular atrophy in both BALB/c and C57/BL6 strains. Excess dietary salt had no significant effect on the severity of pit cell hyperplasia. No dysplasia was found in any of the groups. In C57/BL6 mice, CA IX-deficiency was associated with gastric submucosal inflammation. The results indicate that CA IX-deficiency provides a useful model to study the mechanisms of gastric morphogenesis and epithelial integrity. Further studies are needed to see whether CA IX has a role in the regulation of immune response. The findings suggest that although CA IX deficiency is not a tumor-promoting factor per se, it induces glandular atrophy in the body mucosa, a lesion which is considered to be a preneoplastic alteration in the stomach. PMID- 16245157 TI - Assessment of nematode resistance in wheat transgenic plants expressing potato proteinase inhibitor (PIN2) gene. AB - Serine proteinase inhibitors (IP's) are proteins found naturally in a wide range of plants with a significant role in the natural defense system of plants against herbivores. The question addressed in the present study involves assessing the ability of the serine proteinase inhibitor in combating nematode infestation. The present study involves engineering a plant serine proteinase inhibitor (pin2) gene into T. durum PDW215 by Agrobacterium-mediated transformation to combat cereal cyst nematode (Heterodera avenae) infestation. Putative T(0) transformants were screened and positive segregating lines analysed further for the study of the stable integration, expression and segregation of the genes. PCR, Southern analysis along with bar gene expression studies corroborate the stable integration pattern of the respective genes. The transformation efficiency is 3%, while the frequency of escapes was 35.71%. chi(2) analysis reveals the stable integration and segregation of the genes in both the T(1) and T(2) progeny lines. The PIN2 systemic expression confers satisfactory nematode resistance. The correlation analysis suggests that at p < 0.05 level of significance the relative proteinase inhibitor (PI) values show a direct positive correlation vis-a-vis plant height, plant seed weight and also the seed number. PMID- 16245158 TI - Expression of the small peptide GLP-1 in transgenic plants. AB - Glucagon-like peptide 1 (GLP-1) has great potential in diabetes therapy. In order to accumulate GLP-1 in endosperm tissue of rice, a codon-optimized GLP-1 (mGLP-1) synthetic gene was directly expressed under the control of rice storage protein glutelin GluB-1 promoter in transgenic rice plants. Unexpectedly, neither the transcripts nor the transgene products were detected in the seeds of regenerated plants. Furthermore, transcripts of GluB-1 gene in these transgenic plants were not detected, and small interfering RNAs (siRNAs) corresponding to the transgene were detected. These results indicated that the expression of mGLP-1 was silenced by co-suppression in rice transgenic seeds. To avoid silencing, mGLP-1 was fused to GFP with or without self-processing 2A sequence, and introduced into rice plants. Both chimeric genes were highly expressed in these transgenic rice seeds, indicating that gene silencing could be avoided by changing the transgene components. Furthermore, the fusion protein containing the 2A sequence were processed into GFP-2A and mGLP-1 peptides with the efficiency of more than 80%, but the processed mGLP-1 peptides were not detected. Lack of accumulation of mGLP 1 may be explained by proteolytic digestion in the cytoplasm. PMID- 16245159 TI - Cryopreserved morulae can be used to efficiently generate germline-transmitting chimeras by blastocyst injection. AB - The production of chimeric mice is a complex process, requiring the careful coordination of tissue culture cell growth, production of a large number (30-75) of competent blastocysts and the availability of appropriately timed pseudo pregnant female mice. Failure at any of these steps can impinge upon the rapid production of chimeras. One potential improvement for the efficient generation of chimeric mice would be the utilization of cryopreserved embryos suitable for injection. C57Bl/6 morulae were frozen using a standard 2-step protocol with ethylene glycol as the cryopreservation agent. We determined that cryopreserved morulae could thaw, culture to blastocyst stage in KSOM media and survive injection at rates equivalent to control embryos. Cryopreserved morulae were also equivalent to controls at all later stages in the process of production of chimeric mice, including birth rate, percentage chimerism of resulting animals and ability to produce germline progeny. Hence, cryopreservation of morulae for blastocyst injection is a suitable option to enhance the efficiency of chimeric mouse generation. PMID- 16245160 TI - Gonadal effects of a mouse Denys-Drash syndrome mutation. AB - Gonadal effects of the Denys-Drash syndrome (DDS) mutation Wt1(tmT396 )were examined in chimaeric and heterozygous mice. Since the only heterozygote was 41,XXY, Sertoli cell function was assessed by comparison with age-matched control XXY testes. Control XXY Sertoli cells showed immuno-expression of WT1 and androgen receptor (AR) indistinguishable from wild-type (40,XY), but expressed anti-Mullerian hormone (AMH). In contrast, DDS Sertoli cells showed only faint immuno-expression of WT1 and did not express AR or AMH. While XY<-->XY DDS chimaeras were male, XX<-->XY chimaeras were predominantly female. In the rare XX<-->XY DDS males the Sertoli cell lineage was largely derived from Wt1 mutant XY cells. We conclude that DDS mutant cells can form Sertoli cells, that the dominant mutation does not cause male sex reversal in mice but distorts the sex ratio of XX<-->XY chimaeras, and that there may be a link between WT1, AMH and AR expression by Sertoli cells in vivo. PMID- 16245161 TI - Ectopic expression of an FT homolog from citrus confers an early flowering phenotype on trifoliate orange (Poncirus trifoliata L. Raf.). AB - Citrus FT (CiFT) cDNA, which promoted the transition from the vegetative to the reproductive phase in Arabidopsis thaliana, when constitutively expressed was introduced into trifoliate orange (Poncirus trifoliata L. Raf.). The transgenic plants in which CiFT was expressed constitutively showed early flowering, fruiting, and characteristic morphological changes. They started to flower as early as 12 weeks after transfer to a greenhouse, whereas wild-type plants usually have a long juvenile period of several years. Most of the transgenic flowers developed on leafy inflorescences, apparently in place of thorns; however, wild-type adult trifoliate orange usually develops solitary flowers in the axils of leaves. All of the transgenic lines accumulated CiFT mRNA in their shoots, but there were variations in the accumulation level. The transgenic lines showed variation in phenotypes, such as time to first flowering and tree shape. In F(1) progeny obtained by crossing 'Kiyomi' tangor (C. unshiu x sinensis) with the pollen of one transgenic line, extremely early flowering immediately after germination was observed. The transgene segregated in F(1) progeny in a Mendelian fashion, with complete co-segregation of the transgene and the early flowering phenotype. These results showed that constitutive expression of CiFT can reduce the generation time in trifoliate orange. PMID- 16245162 TI - A model transgenic cereal plant with detoxification activity for the estrogenic mycotoxin zearalenone. AB - Zearalenone (ZEN) is an estrogenic mycotoxin produced by the necrotrophic cereal pathogen Fusarium graminearum. This mycotoxin is detoxified by ZHD101, a lactonohydrolase from Clonostachys rosea, or EGFP:ZHD101, its fusion to the C terminus of an enhanced green fluorescence protein. We previously showed that egfp:zhd101 is efficiently expressed in T(0) leaves of rice. In this study, we assessed the feasibility of in planta detoxification of the mycotoxin using progeny. When protein extract from T(1) leaves was incubated with ZEN, the amount of the toxin decreased significantly as measured by HPLC. ZEN degradation activity was also detected in vivo in transgenic T(2) seeds. These results suggest that zhd101 can be exploited as an efficient and cost-effective system for protection of important cereals that are more susceptible to the pathogen (e.g., wheat and maize) from contamination with the estrogenic mycotoxin. PMID- 16245163 TI - Development of transgenic mice expressing calcitonin as a beta-lactoglobulin fusion protein in mammary gland. AB - Expression of foreign proteins in mammalian milk is becoming a widespread strategy for high-level production of recombinant pharmaceuticals, especially those with the most complex post-translational modifications. A milk-specific ovine beta-lactoglobulin (oBLG) promoter was used to drive expression of recombinant calcitonin in mouse milk. A gene construct was generated, consisting of 10.7 kbp of the oBLG gene including its promoter and 3' flanking region with the calcitonin coding sequences inserted in-frame into the oBLG fifth exon. After microinjection, six founder mice transmitted the transgene to their progeny. RT PCR confirmed mammary-gland specific expression of recombinant mRNA in most transgenic mice and Western blot analysis confirmed expression of chimeric protein. Calcitonin can thus be expressed under the oBLG promoter and regulatory elements in a mammary-gland specific manner. PMID- 16245164 TI - Strain differences in superovulatory response, embryo development and efficiency of transgenic rat production. AB - The differences between rat strains in superovulation response, in vitro and in vivo development of preimplantation embryos and overall transgenic efficiency was studied. The protocols for induction of superovulation using single injections of pregnant mare's serum gonadotropin (PMSG) or minipumps with follicle stimulating hormone (FSH) were compared in Lewis (LEW), Wistar-Kyoto (WKY), and stroke-prone spontaneously hypertensive rats (SHRSP) or Sprague-Dawley (SD) and Wistar rats as representative inbred or outbred strains, respectively. The percentage of mated animals with positive superovulatory response was similar in all strains (60.0 100%). The mean number of ova per donor was not dependent on the kind of hormonal treatment used within each rat strain. In general, females from outbred SD and Wistar rats were more responsive to hormonal treatments than animals from inbred rat strains. In addition, SD female rats produced a significantly higher number of embryos per female in response to PMSG-treatment compared to all other strains. Between the inbred strains, SHRSP was the most effective for superovulation. In vitro development of intact zygotes to the blastocyst stage was not different between SD, Wistar and SHRSP rats. In contrast, in vitro development of WKY zygotes was significantly less efficient than in other strains. However, 2-cell stage embryos in vivo produced from SD, SD x Wistar and WKY animals showed no difference in competence to develop to blastocyst stage in vitro. The proportion of offspring developing after oviduct transfer of intact zygotes was similar in all strains (44.0-56.4%) with the exception of WKY rats (35.9%). We also compared the survival rate after injection, ability of manipulated zygotes to develop to term and overall transgenic efficiency in various rat strains. SD and SHRSP zygotes survived after microinjection better than the WKY and Lewis zygotes. No differences were found in the efficiency of transgene integration per newborn in different strains ranging from 5.7 to 16.7%. The results of this study demonstrate that different rat strains have varying responses to superovulation, sensitivity to microinjection, capability to develop in vitro until blastocyst stage or in vivo to term after transfer to foster mothers. Despite these differences all studied strains can be used for efficient transgenic rat production. PMID- 16245165 TI - Nicotianamine over-accumulation confers resistance to nickel in Arabidopsis thaliana. AB - Nicotianamine is a methionine derivative involved in iron homeostasis, able to bind various other metals in vitro. To investigate its role in vivo, we expressed a nicotianamine synthase cDNA (TcNAS1) isolated from the polymetallic hyperaccumulator Thlaspi caerulescens in Arabidopsis thaliana. Transgenic plants expressing TcNAS1 over-accumulated NA, up to 100-fold more than wild type plants. Furthermore, increased NA levels in different transgenic lines were quantitatively correlated with increased nickel tolerance. The tolerance to nickel is expressed at the cellular level in protoplast experiments and is associated with an increased NA content. We have also shown that the most NA-over accumulating line showed a high tolerance to nickel and a significant Ni accumulation in the leaves when grown on nickel-contaminated soil. Our results highlight a new potential role for nicotianamine in heavy metal tolerance at the cellular but also at the whole plant level, easily transposable to a non-tolerant non-hyperaccumulator species. These results open new perspectives for the modulation of nicotianamine content in plants for phytoremediation. PMID- 16245166 TI - Crop-to-crop gene flow using farm scale sites of oilseed rape (Brassica napus) in the UK. AB - From 2000-2003 a range of Farm Scale Evaluation (FSE) trials were established in the UK to assess the effect of the release and management of herbicide tolerant (HT) crops on the abundance and diversity of farmland wildlife compared with their conventionally managed non-GM-equivalents. The objective of this research project was to investigate gene flow within the winter (WOSR) and spring oilseed rape (SOSR) FSE trials and to develop a statistical model for the prediction of cross-pollination frequency that can be used to evaluate current separation distance guidelines. Seed samples were collected from the non-GM half of the trial sites and were tested for evidence of cross-pollination from the GM HT halves using a quantitative PCR assay specific to the HT (bar) gene. Rates of cross-pollination were found to decrease rapidly with increasing distance from the GM source. The quantitative data were subjected to statistical analysis and a two-step model was found to provide the best fit for the data. Significant differences were found between the results for WOSR, SOSR and varietal association (VA) crops. The model predicted that the %GM content (including upper 95% confidence limits) of a sample taken at a distance of 50 m away from the GM source would be 0.04% (0.84%) for WOSR, 0.02% (0.39%) for SOSR, 0.77% (21.72%) for WOSR VA and 0.37% (5.18%) for SOSR VA. The data and models presented here are discussed in the context of necessary separation distances to meet various possible thresholds for adventitious presence of GM in OSR. PMID- 16245168 TI - Assessing the transfer of genetically modified DNA from feed to animal tissues. AB - In Europe, public and scientific concerns about the environmental and food safety of GM (Genetically Modified) crops overshadow the potential benefits offered by crop biotechnology to improve food quality. One of the concerns regarding the use of GM food in human and animal nutrition is the effect that newly introduced sequences may have on the organism. In this paper, we assess the potential transfer of diet-derived DNA to animal tissues after consumption of GM plants. Blood, spleen, liver, kidney and muscle tissues from piglets fed for 35 days with diets containing either GM (MON810) or a conventional maize were investigated for the presence of plant DNA. Only fragments of specific maize genes (Zein, Sh-2) could be detected with different frequencies in all the examined tissues except muscle. A small fragment of the Cry1A(b) transgene was detected in blood, liver, spleen and kidney of the animals raised with the transgenic feed. The intact Cry1A(b) gene or its minimal functional unit were never detected. Statistical analysis of the results showed no difference in recovery of positives for the presence of plant DNA between animals raised with the transgenic feed and animals raised with the conventional feed, indicating that DNA transfer may occur independently from the source and the type of the gene. From the data obtained, we consider it unlikely that the occurrence of genetic transfer associated with GM plants is higher than that from conventional plants. PMID- 16245167 TI - Mammary specific transgenic over-expression of insulin-like growth factor-I (IGF I) increases pig milk IGF-I and IGF binding proteins, with no effect on milk composition or yield. AB - IGF-I regulates lactation by stimulating mammary mitogenesis, inhibiting apoptosis, and partially mediating the effects of growth hormone on lactogenesis. Herein, lactation performance during first and second parity was assessed in transgenic swine (TG) that over-expressed human IGF-I in milk under the control of the bovine alpha-lactalbumin promoter, regulatory regions and signal peptide coding sequence. Milk samples were collected throughout lactation (farrowing to d24) from TG sows and non-transgenic littermates (CON) and IGF-I, IGF-II, and IGFBP determined. Colostral (<24 h postpartum) IGF-I content was 26-fold greater (p<0.001) in TG sows (949+/- 107 microg/L; range 228-1,600 microg/L) than CON (36+/-17.8 microg/L) and was 50- to 90-fold greater (p< 0.001) in mature milk (d2 24 postpartum). There was no effect of parity on milk IGF-I content. Milk IGF-II concentration was unaffected by IGF-I over-expression. Low molecular weight IGFBP (IGFBP-2 and -5) in the milk of TG sows were higher (p=0.02) than CON in the early postpartum period, but did not differ in mature milk. Milk yield, determined by weigh-suckle-weigh, was similar in TG and CON as was litter weight gain. Milk nutrient composition was not significantly affected by IGF over expression. Thus, mammary specific transgenic over-expression of IGF-I significantly increased milk IGF-I and IGFBP content, but did not impact lactation performance in swine. PMID- 16245170 TI - Agroinfiltration as a tool for transient expression of cre recombinase in vivo. AB - Agroinfiltration was used to express transiently cre recombinase from bacteriophage P1 in planta. Activation of gfp expression after cre-mediated excision of a bar intervening sequence served as a marker to monitor site specific recombination events in lox-target N. benthamiana plants. Gfp expressing regenerants from A. tumefaciens infiltrated leaves were obtained with an efficiency of about 34%. In 20% of the regenerants bar gene excision was due to the expression of stably integrated cre gene, whereas in 14% of plants site specific recombination was a consequence of transient cre expression. Phenotypic and molecular data indicated that the recombined state has been transferred to the T(1 )generation. These results demonstrate the suitability of agroinfiltration for the expression of cre recombinase in vivo. PMID- 16245171 TI - Response surface analysis of solid state growth of Pleurotus ostreatus mycelia utilizing whey permeate. AB - A novel approach to utilizing whey permeate, the cultivation of mycelia of the edible mushroom Pleurotus ostreatus, is introduced. Response surface analysis (RSA) was successfully applied to determine the combination of substrate concentration, temperature and pH that would result in a maximal mycelial extension rate under solid state cultivation. The conditions to maximize the mycelial extension rate were predicted to be 44 g lactose l(-1), pH 6.0 and 24.2 degrees C. Subsequent verification of these levels agreed with model predictions. PMID- 16245169 TI - Transgenic tobacco plants expressing a dimeric single-chain variable fragment (scfv) antibody against Salmonella enterica serotype Paratyphi B. AB - Transgenic tobacco plants were produced that express an anti-Salmonella enterica single-chain variable fragment (scFv) antibody that binds to the lipopolysaccharide (LPS) of S. enterica Paratyphi B. The coding sequence of this scFv was optimized for expression in tobacco, synthesized and subsequently placed behind three different promoters: an enhanced tobacco constitutive ubiquitous promoter (EntCUP4), and single- and double-enhancer versions of the Cauliflower Mosaic Virus 35S promoter (CaMV 35S). These chimeric genes were introduced into Nicotiana tabacum cv. 81V9 by Agrobacterium-mediated transformation and 50 primary transgenic (T(0)) plants per construct were produced. Among these plants, 23 were selected for the ability to express active scFv as determined by enzyme linked immunosorbent assay (ELISA) using S. enterica LPS as antigen. Expanded bed adsorption-immobilized metal affinity chromatography (EBA-IMAC) was used to purify 41.7 mug of scFv/g from leaf tissue. Gel filtration and surface plasmon resonance (SPR) analyses demonstrated that the purified scFv was active as a dimer or higher-order multimer. In order to identify T(1) plants suitable for development of homozygous lines with heritable scFv expression, kanamycin resistance segregation analyses were performed to determine the number of T-DNA loci in each T(0) plant, and quantitative ELISA and immunoblot analyses were used to compare expression of active and total anti-Salmonella scFv, respectively, in the T(1) generation. As S. enterica causes millions of enteric fevers and hundreds of thousands of deaths worldwide each year, large-scale production and purification of this scFv will have potential for uses in diagnosis and detection, as a therapeutic agent, and in applications such as water system purification. PMID- 16245172 TI - Expression of the cercosporin transporter, CFP, in tobacco reduces frog-eye lesion size. AB - The cercosporin Major Facilitator Superfamily (MFS) transporter, CFP, under the control of the CaMV 35S promoter, was introduced into the Xanthi cultivar of tobacco by Agrobacterium-mediated transformation. CFP(+) transgenic plants were physically indistinguishable from non-transgenic Xanthi and progressed normally through growth to seed set. Accumulation of CFP in the leaf membrane fraction of CFP(+ )transgenic plants was associated with decreased cercosporin phytotoxicity. Frog-eye leaf lesions on CFP(+ )transgenic plants infected with Cercospora nicotianae conidia were smaller but were similar in number to those on non transgenic plants. We conclude that transgenic expression of CFP may have relevance for a disease control strategy in Cercospora-plant pathosystems where cercosporin is implicated in pathogen virulence. PMID- 16245173 TI - Lactic acid bacteria biomass monitoring in highly conductive media by permittivity measurements. AB - Recent technological improvements have extended the application range of permittivity biomass measurements to bacterial fermentations in highly conductive media. With Lactobacillus casei, the effective biomass detection sensitivity of the FOGALE Biomass System is around 0.2 g/l (0.01 pF/cm). Fermentations growth kinetics of Lactobacillus casei can be recorded with good reproducibility and accuracy despite the high medium conductivity varying between 15 and 75 mS/cm, and the low cell concentration (<6 g/l). PMID- 16245174 TI - Expression and purification of biologically active rat bone morphogenetic protein 4 produced as inclusion bodies in recombinant Escherichia coli. AB - Rat bone morphogenetic protein-4 (rBMP-4) cDNA was cloned from rat osteoblasts by RT-PCR and expressed in E. coli. Monomeric, dimeric and polymeric forms of recombinant rat BMP-4 (rrBMP-4) were obtained from inclusion bodies after solubilization with urea. The dimer was separated from the remaining polymer and host cell contaminants using size exclusion chromatography. Furthermore, purified rrBMP-4 was stabilized at low urea concentration (40 mM) and at pH 8.5 through the addition of bovine serum albumin. Both, rrBMP-4 dimer and polymer were biologically active as tested by the induction of alkaline phosphatase activity in MC3T3-E1 cells. PMID- 16245175 TI - Retention of browning compounds by yeasts involved in the winemaking of sherry type wines. AB - Wine model solutions were used to study the ability of dehydrated yeasts to retain the brown products formed in the reaction between (+)-catechin and acetaldehyde. Saccharomyces cerevisiae races capensis and bayanus, two typical flor yeasts involved in the biological aging of sherry wines, had a higher capacity to retain coloured compounds than S. cerevisiae fermentative yeast. Of the flor yeasts, capensis exhibited a higher colour reduction capacity than bayanus. Such differences may account for the different rate at which browning compounds are removed at different times of year during the biological aging of wines. PMID- 16245176 TI - Protein-coated microcrystals for use in organic solvents: application to oxidoreductases. AB - Protein-coated microcrystals (PCMC), a biocatalyst preparation previously demonstrated to display substantially increased transesterification activity of proteases and lipases in organic solvents when compared to their as received counterparts [Kreiner M, Moore BD, Parker MC (2001) Chem. Commun. 12:1096--1097], was applied to oxidoreductases. Horse liver alcohol dehydrogenase (HLADH), catalase (CAT), soybean peroxidase and horseradish peroxidase were immobilised onto K(2)SO(4) crystals and dehydrated in a 1-step process, resulting in PCMC. These PCMC preparations showed enhanced activity in different organic solvents in most types of reactions tested. The highest activation was observed with HLADH (50-fold as active as enzyme as received) and CAT (25-fold). PMID- 16245177 TI - The use of phase inversion temperature (PIT) microemulsion technology to enhance oil utilisation during Streptomyces rimosus fed-batch fermentations to produce oxytetracycline. AB - The use of a rapeseed oil emulsion feed, produced by a phase inversion temperature (PIT) process, produced more biomass, gave a 3-fold increase in oil utilisation and a higher oxytetracycline titre but a higher residual oil concentration when compared to a conventional fed-batch Streptomyces rimosus process fed with crude rapeseed oil. Importantly, microbial utilisation of the surfactant was confirmed for the first time. PMID- 16245178 TI - Characterisation of hydrogel gel swelling by molecular exclusion. AB - A facile method for the characterization of hydrogel swelling is described which is based on the determination of changes in the liquid phase concentration of an excluded tracer as gel swells in a constant volume system. The utility of this approach is demonstrated with two responsive hydrogel preparations, one where swelling is influenced by system pH, the other by changes in specific solute concentration. PMID- 16245179 TI - Controllable regioselective acylation of rutin catalyzed by enzymes in non aqueous solvents. AB - An efficient route to synthesize 3''- and 4'''-vinyl rutin esters has been developed by enzyme-catalyzed regioselective acylation of rutin with divinyl dicarboxylates in organic media. Alkaline protease from Bacillus subtilis provided 3''-O-substituted vinyl rutin esters in pyridine, and Novozym 435 gave 4'''-O-substituted vinyl rutin esters in tert-butanol. PMID- 16245180 TI - Synthesis, expression and purification of a type of chlorotoxin-like peptide from the scorpion, Buthus martensii Karsch, and its acute toxicity analysis. AB - A gene, rBmK Cta, encoding a chlorotoxin-like peptide from the scorpion, Buthus martensii Karsch, was synthesized according to the sequence optimized for codon usage in Escherichia coli and was expressed in E. coli BL21 (DE3) using a pExSecI expression system in which the IgG-binding domain-ZZ of protein A is fused to the N-terminal of rBmK CTa. The fusion protein, ZZ-rBmK CTa, was expressed in soluble form (7.8 mg l(-1)) and was purified to give a single band on SDS-PAGE. The domain-ZZ of fusion protein ZZ-rBmK CTa was removed by cleavage of an Asn-Gly peptide bond with hydroxylamine. The rBmK CTa was separated from the IgG-binding moiety by a second passage through the IgG affinity column. Western blot analysis demonstrated that this protein was rBmK CTa. Acute toxicity assay in mice demonstrated that the rBmK CTa had an LD(50) value of 4.3 mg kg(-1). PMID- 16245181 TI - Surfactin C inhibits the lipopolysaccharide-induced transcription of interleukin 1beta and inducible nitric oxide synthase and nitric oxide production in murine RAW 264.7 cells. AB - The anti-inflammatory activity of the surfactin C derived from Bacillus subtilis isolate was investigated in lipopolysaccharide (LPS, 1 microg ml(-1))-treated mouse RAW 264.7 cells. LPS increased mRNA transcription of cyclooxygenase (COX) 2, interleukin (IL)-1beta and inducible nitric oxide synthase (iNOS). However, surfactin C at 50 microg ml(-1 )inhibited the LPS-induced increase in the transcription of IL-1beta and iNOS and nitric oxide (NO) production in a dose dependent manner. PMID- 16245182 TI - Antagonism of transforming growth factor-Beta signaling inhibits fibrosis-related genes. AB - In the fibrotic process, the transforming growth factor-beta1 (TGF-beta1)/Smad3 (Sma- and Mad-related protein 3) signaling plays a central role. To screen for antagonists of TGF-beta1/Smad3 signaling and to investigate their effects on the genes related to fibrosis, we construct a molecular model with a luciferase reporter gene. Results showed that both SB-431542 [4-(5-benzo[1,3]dioxol-5-yl-4 pyridin-2-yl-1H-imidazol-2-yl)-benzamide] and small interference RNA (siRNA) against Smad3 could dose-dependently suppress the reporter gene. More importantly, they both significantly inhibited the expression of plasminogen activator inhibitor-type 1 (PAI-1) and type I collagenalpha1 (Col Ialpha1) genes in rat hepatic stellate cells. Thus, SB-431542 and Smad3/siRNA may be potential therapeutics for fibrosis. PMID- 16245183 TI - Decalactone production by Yarrowia lipolytica under increased O2 transfer rates. AB - Yarrowia lipolytica converts methyl ricinoleate to gamma-decalactone, a high value fruity aroma compound. The highest amount of 3-hydroxy-gamma-decalactone produced by the yeast (263 mg l(-1)) occurred by increasing the k(L)a up to 120 h(-1) at atmospheric pressure; above it, its concentration decreased, suggesting a predominance of the activity of 3-hydroxyacyl-CoA dehydrogenase. Cultures were grown under high-pressure, i.e., under increased O(2) solubility, but, although growth was accelerated, gamma-decalactone production decreased. However, by applying 0.5 MPa during growth and biotransformation gave increased concentrations of dec-2-en-4-olide and dec-3-en-4-olide (70 mg l(-1)). PMID- 16245184 TI - Yeast extract from Express Five serum-free medium contains factors at about 35 kDa, essential for growth of Trichoplusia ni insect cells. AB - The yeast extract (of unknown origin) present in the commercially available serum free medium 'Express Five' contains factors ('yeast extract factors') up to 35 kDa which are essential for growth of Trichoplusia ni insect cells. A yeast extract brand lacking these components could not support growth of T. ni cells. However, cell proliferation was restored by adding chromatographic fractions containing the yeast extract factors. The yeast extract factors were not solely responsible for the growth enhancing effect of yeast extract but some other components, which seem to be generally present in yeast extracts, are also required for T. ni proliferation. PMID- 16245185 TI - Direct FTIR assay of streptomycin in agar. AB - Streptomycin titres in samples of agar media on which various species of streptomycetes were cultured were obtained by Fourier Transform Infra Red (FTIR) spectroscopy. Titres were directly comparable to those obtained by bioassay based on Bacillus subtilis inhibition. Analysis by this method could be used to facilitate the isolation of high level antibiotic-producing mutants. PMID- 16245186 TI - Construction of a novel extracellular matrix using a new genetically engineered epidermal growth factor fused to IgG-Fc. AB - The design of artificial extracellular matrices has attracted much attention in tissue engineering as well as in cell biology research. An immobilized recombinant epidermal growth factor (EGF), fused to an immunoglobulin G (IgG) Fc region (abbreviated as EGF-Fc) has been constructed. Mouse fibroblast Swiss 3T3 cells adhered both to EGF-Fc-coated and collagen-coated surfaces. Phosphorylation of EGF receptor in A431 cells was induced by immobilized EGF-Fc as well as soluble EGF. Immobilized EGF-Fc continuously activated mitogen-activated protein kinase (MAPK) in A431 cells whereas MAPK activation induced by soluble EGF decreased rapidly with time. The cytoskeleton of A431 cells adhering onto immobilized EGF-Fc was filopodia whereas that of the cells adhering onto collagen in the presence of soluble EGF was lammellipodia. PMID- 16245189 TI - Factors associated with ideation and acts of deliberate self-harm among 18-year old boys. A prospective 10-year follow-up study. AB - BACKGROUND: The aims of the study were: (1) to identify the prevalence of ideation and acts of deliberate self-harm in 18-year-old boys; (2) to report the use of mental health services among these boys; (3) to identify risk factors assessed at age 8 and (4) to identify associated cross-sectional factors at age 18 of self-reported ideation and acts of deliberate self-harm. METHODS: The participants in this community-based 10-year follow-up study consisted of 2,348 boys born during 1981. At baseline, three informant sources were used: parents, teachers and the children themselves. The questionnaires used were the Rutter Parents Scale, the Rutter Teacher Scale and the Children's Depressive Inventory. At follow-up, boys' psychopathology, adaptive functioning and substance use were studied using the Young Adult Self-Report questionnaire. Further questions about demographic factors and life events were added at both stages. RESULTS: The prevalence of ideation of deliberate self-harm was 4.0%, and of acts 2.2%, during the preceding 6 months. Of the subjects who reported ideation, 9.7%, and of those who reported acts, 16.0%, had used mental health services during the previous year. Self-reported depressive symptoms at age 8 predicted ideation and acts of deliberate self-harm 10 years later. Suicidal boys' problems in many life areas were linked with suicidal tendency through psychiatric symptoms, especially anxious/depressed symptoms and aggressive behaviour. CONCLUSIONS: The connection between self-reported depressive symptoms at age 8 and ideation and acts of deliberate self-harm at age 18 is a good reason to already pay special attention to children's own intimations of distress in elementary school. It is likely that an effective way to prevent suicidality among adolescents is to search for and treat psychiatric problems among young people. Self-reported screening questionnaires used, e.g. in school healthcare, may provide an opportunity to recognize these problems. PMID- 16245191 TI - [Meditation in El Masnou. The resident has come. Nobody knows how it was]. PMID- 16245190 TI - Burden of care and general health in families of patients with schizophrenia. AB - BACKGROUND: De-institutionalization of psychiatric patients has led to a greater emphasis on family management in the community, and family members are often overwhelmed by the demands that caring for a patient with schizophrenia involves. Most studies of family burden in schizophrenia have taken place in developed countries. The current study examined family burden and its correlates in a regional area of a medium income country in South America. METHOD: Sixty-five relatives of patients with schizophrenia who were attending a public mental health out-patient service in the province of Arica, Chile, were assessed on Spanish versions of the Zarit Caregiver Burden Scale and SF-36 Health Survey (SF 36). RESULTS: Average levels of burden were very high, particularly for mothers, carers with less education, carers of younger patients and carers of patients with more hospitalisations in the previous 3 years. Kinship and number of recent hospitalisations retained unique predictive variance in a multiple regression. Burden was the strongest predictor of SF-36 subscales, and the prediction from burden remained significant after entry of other potential predictors. CONCLUSIONS: In common with families in developed countries, family members of schizophrenia patients in regional Chile reported high levels of burden and related functional and health impact. The study highlighted the support needs of carers in contexts with high rates of poverty and limited health and community resources. PMID- 16245192 TI - [Light exposure, lipofuscin and age-related macular degeneration]. PMID- 16245193 TI - [Oral clarithromycin for saccular endophthalmitis treatment]. PMID- 16245194 TI - [Comparison of three methods of inducing chronic elevation of intraocular pressure in the pig (experimental glaucoma)]. AB - PURPOSE: To compare three methods of inducing experimental glaucoma in the adult pig, based on achieving chronic elevation of the intraocular pressure (IOP). METHODS: A total of 16 adult pigs were used in the present study. In all instances, the right eye was used as control and the left eye as the experimental eye. The animals were divided into three groups: 1) 3 pigs in which 3 episcleral veins were cauterized (experimental period = 21 weeks); 2) 6 animals in which the anterior chamber of the eye was injected with a solution containing latex fluorospheres (experimental period = 11 weeks); and 3) 4 pigs in which the anterior chamber of the eye was injected with a solution containing latex fluorospheres plus methylcellulose (experimental period = 11 weeks). RESULTS: The episcleral vein cauterization was the only method which produced a sustained elevation of the IOP throughout most of the experimental period. Moreover, the elevation of the IOP achieved by this method resulted in selective retinal ganglion cell (RGC) loss that affected mainly the mid-peripheral and peripheral retina and caused an increase in the mean soma area of the remaining RGCs. CONCLUSIONS: Cauterization of the episcleral veins resulted in a significant and sustained elevation of the IOP and RGC loss when compared with the other two approaches tested in the present study, which barely modified the pig's RGC distribution. Thus, we conclude that the episcleral vein cauterization is the best of the methods tested to induce experimental glaucoma in the pig. PMID- 16245196 TI - [Intraocular lens power calculation in complicated cases: the "Silguero"method]. AB - PURPOSE: To report a new method for intraocular lens (IOL) calculation in patients treated by corneal refractive surgery, in whom the application of other methods is not possible because of the absence of necessary data. This method is also applicable in other situations where the IOL calculation is difficult and in cases of IOL exchange because of unexpected refractive difficulties. METHODS: We performed phaco-emulsifications, refracting the patient's eye before IOL implantation, using the refraction data in aphakia as the only variable for the application of the formula, which itself is based on the existing correlation between the graduation of glasses (12 mm) and the IOL power at the lens level. The formula has been applied in 3 patients in whom no other calculation method was applicable. RESULTS: After the appropriate intervention, the 3 patients presented a spherical equivalent below 1 diopter. CONCLUSION: We believe this formula is valid in some situations where other formulas cannot be applied because of the absence of the necessary variables. PMID- 16245195 TI - [Two different intra-corneal inlay surgical technique in rabbit eyes]. AB - OBJECTIVE: To compare two different surgical techniques of intracorneal lens implantation. To determine the incidence of corneal and lens complications in two groups of rabbits subjected to such surgery. METHOD: The first group (7 rabbits) initially had a keratectomy performed with the microkeratome following which the corneal flap was sutured to the stromal layer. In the second group (7 rabbits), a pocket-shaped stromal dissection was performed manually. Both groups of rabbits had a secondary lens implantation in the corneal inlay. The follow up period was of 2 months duration. RESULTS: The incidence of corneal edema in the first month was higher in group 1 (71.4%) than in group 2 (14.3%) (p = 0.0155); during the second month, the incidence was 42.9% in group 1 and 28.6% in group 2 (p= 0.2927). Flap dislocation and retraction of the flap was seen in 4 of the 7 rabbits in group 1, whereas no complication of this type was observed in the manual pocket group (group 2). The incidence of lens subluxation was 28.6% in group 1 and 0% in group 2. Finally, there was a 71.4% (5/7) loss of the intracorneal lens in the rabbits treated with the microkeratome (group 1) while only 1 of 7 rabbits (14.3%) lost the lens in group 2 (P = 0.0155). DISCUSSION: Eyes in which the corneal pocket shape was made by manual stromal dissection had a higher stability of the lens and less post-operative corneal complications. PMID- 16245197 TI - [Laser photocoagulation in idiopathic polypoidal choroidal vasculopathy. Over one year follow-up]. AB - PURPOSE: Long term assessment of the evolution of Idiopathic Polypoidal Choroidal Vasculopathy (IPCV) in symptomatic patients treated by laser photocoagulation. METHODS: We carried out a retrospective study involving seven eyes in six patients with visual impairment, affected by IPCV, confirmed by indocyanine green angiography. All were treated by laser photocoagulation and monitored during a period of at least one year. The controls were evaluated during the first, third, sixth and twelfth month, in order to assess whether it was necessary to repeat the treatment. RESULTS: Of the six patients with IPCV lesions, two were men and four were women (1:2) aged between 58 and 81 years (mean = 73 years), and monitored for a time interval of 18 and 75 months (mean = 39 months). The average initial visual acuity was 0.25, and the final was 0.15. All the patients had the other eye also affected: three had a macular disciform scar, secondary to the previous haemorrhagic episodes, two had asymptomatic polyps and in one patient we defined active bilateral IPCV which was treated by laser photocoagulation. In two eyes it appeared as a macular haemorrhage, in four eyes as a serosanguinous retinal pigment epithelial (RPE) detachment and in one eye as a serous RPE detachment. Three eyes suffered a relapse which required further laser photocoagulation treatment. CONCLUSIONS: Laser photocoagulation is a palliative treatment for IPCV. Final vision achieved is poor and relapse is frequent. Other procedures should be used to treat this disease. PMID- 16245198 TI - [Acute zonal outer occult retinopathy]. AB - CASE REPORT: In May 2001 a 10-year-old boy, examined 3 months after the development of a large temporal scotoma in the right eye, was diagnosed with photopsia. The fundus appeared normal, visual field examination revealed a large temporal defect and, in July 2001, electroretinography (ERG) was nonrecordable. The left eye was normal. The field defects and ERG abnormality remained 4 years later, and the fundus developed a sharply demarcated zone of atrophy of the RPE. DISCUSSION: We report a typical case of acute zonal occult outer retinopathy (AZOOR). Although rare, its symptoms are variable, and its shape is not characteristic, making it difficult to make the correct diagnosis. PMID- 16245199 TI - [Tortuous retinal vessels in neurofibromatosis type 1]. AB - CASE REPORT: We report a 4-year-old patient with neurofibromatosis type 1 (NF-1) who had tortuous retinal vessels of first, second and third order which ended in a corkscrew appearance. DISCUSSION: Vascular retinal abnormalities can be present in patients with NF-1. PMID- 16245200 TI - [Acute concomitant esotropia and hysterical amblyopia]. AB - CASE REPORT: The authors present the case of a 26-year-old woman, in her 29th week of pregnancy, who developed intermittent esotropia and diplopia. Six months later, the process was associated with visual loss and an accommodative spasm which did not improve with cycloplegia and refraction. The symptoms remained unchanged two years later. DISCUSSION: Acute concomitant esotropia may have no obvious underlying cause, but it has been associated with disruption of fusion, myopia, neurological problems and decompensation of a pre-existing phoria or monofixation syndrome. A physical or emotional debilitating illness may precede the onset of the problem. In our case, a neurologic origin was excluded and a diagnosis of hysterical neurosis and acute concomitant esotropia with hysterical amblyopia or functional visual loss was established. PMID- 16245201 TI - [A case of triamcinolone-induced pseudo-endophthalmitis after vitrectomy]. AB - CASE REPORT: Vitrectomy with vitreous injection of triamcinolone in a patient suffering cystoid macular edema, secondary to a lens fragment luxation after subluxated cataract phacoemulsification, was performed. Four days after the surgery she presented because of discomfort and visual loss in the operated eye, and was found to have a 3.4 mm hypopyon. DISCUSSION: We describe the differences between infectious endophthalmitis, toxic endophthalmitis and pseudo endophthalmitis. PMID- 16245202 TI - [Glaucoma secondary to silicone oil in the anterior chamber]. AB - CASE REPORT: A 53-year-old woman, with a history of a right retinal detachment treated by vitrectomy and silicone oil injection, who subsequently had the oil removed. She presented to us with a hypertensive anterior uveitis, which could not be controlled with medical treatment. On examination, micro-bubbles of emulsified silicone oil were evident in the anterior chamber simulating tyndall. A secondary glaucoma to emulsified silicone oil in the anterior chamber was diagnosed, and surgical treatment was proposed. DISCUSSION: Although secondary glaucoma is a frequent complication after oil silicone injection in vitreo retinopathy surgery, the presence of emulsified silicone oil in the anterior chamber simulating tyndall is uncommon. PMID- 16245204 TI - [Saint Otilia (660--Hohenburg, 720)]. PMID- 16245203 TI - [Blindness and other eye diseases in Galdos novels (II)]. PMID- 16245205 TI - Epidermal growth factor receptor in glioblastoma. AB - We compiled the current state of knowledge about the epidermal growth factor receptor (EGFR) in glioblastoma. Glioblastoma is one of the most common primary brain tumours and has an unfavourable prognosis despite aggressive treatment. These factors stimulate new research trials and a recent area of interest of neuro-oncologists is EGFR. This molecule is frequently altered in glioblastoma and constitutes the potential target for therapy. We decided to review the literature on biological structure of that molecule, its biological activity and the role in GBL with potential targeting it in the future neuro-oncological practice. PMID- 16245206 TI - Is there a spatial association between senile plaques and neurofibrillary tangles in Alzheimer's disease? AB - OBJECTIVE: To test the hypothesis that the clusters of senile plaques (SP) and neurofibrillary tangles (NFT) in patients with Alzheimer's disease (AD) are spatially associated as predicted by the 'Amyloid Cascade Hypothesis'. METHODS: The spatial association between the SP and NFT was studied in the cerebral cortex and hippocampus in six cases of sporadic Alzheimer's disease (AD) using contingency tables. The coefficient C7 was used as an index of spatial association while chi-square with correction for continuity was used as a test of significance. RESULTS: In the brain regions analysed, values of C7 were in the range -0.31 to +0.32 but a statistically significant spatial association between SP and NFT was present in only 8/39 (21%) regions. The degree of spatial association between the SP and NFT was similar in different brain regions and did not vary with apolipoprotein e genotype of the patient. However, the magnitude of C7 in a region was positively correlated with the density of the NFT and with the total density of SP and NFT but not with the density of SP alone. CONCLUSION: There was little evidence that SP and NFT were spatially associated except in brain areas with high densities of lesions. The data support the hypothesis that SP and NFT are distributed relatively independently in the cerebral cortex and hippocampus and therefore, could be distinct phenomena in AD. PMID- 16245207 TI - Alzheimer's disease related peptides affected cholinergic receptor mediated poly(ADP-ribose) polymerase activity in the hippocampus. AB - Our previous studies indicated that Alzheimer's disease (AD) related amyloid beta peptide (Abeta) significantly altered muscarinic cholinergic receptor (mChR) signaling on the level of G protein regulated phospholipase C (PLC) leading to the lower formation of inositol-1,4,5-triphosphate (IP3) and diacylglycerol (DAG). Recent studies indicated that poly (ADP-ribose) polymerase-1 (PARP-1) is a new nuclear target in signal transduction pathway in the brain. In this study the effect of Abeta 25-35 (25 microM) and non-Abeta component of Alzheimer's disease amyloid (NAC, 10 microM) on mChR-dependent signaling to PARP-1 was determined. PARP-1 activity was estimated radiochemically using egzogenous substrate adenine[14C]NAD. The results showed that the non hydrolysable agonist of mChR, carbachol (1 mM) together with GTP(g)S (100 microM) stimulated PARP-1 activity in the hippocampus by about 100%. TMB-8, inhibitor of IP3 receptor decreased PARP-1 activation evoked by carbachol/GTP(g)S. Stimulation of mChR did not lead to free radicals generation but activate PARP-1 through IP3/Ca2+ regulated processes. This cholinergic receptor dependent PARP-1 activation was abolished by Abeta and NAC peptide. These toxic peptides themselves significantly stimulated PARP-1 activity by free radicals mediated DNA damage. These data indicated that Abeta and NAC peptide affected mChR-dependent signal transduction to PARP-1 probably through free radicals evoked inhibition of IP3 formation by phospholipase C. PMID- 16245208 TI - The analysis of the intra-surgical view of the obstetric brachial plexus palsy. AB - The surgical intervention is necessary in about 20% of all cases of the perinatal brachial plexus palsy. In this study the intra-surgical view and the applied microsurgical techniques were analysed. The clinical material consisted of 49 children with the obstetric brachial plexus palsy treated operatively, on which the following changes were found intra-surgically: compression of the brachial plexus in 21 cases and injuries with discontinuity of the elements of the brachial plexus in 28 cases. The following surgical procedures were performed: neurolysis - 31 cases, direct neurorrhaphy - 7 cases, 2 - 4 sural nerve grafting from 2 to 3 cm - 4 cases, extra-anatomical extra-plexual reconstruction - 6 cases, extra-anatomical intra-plexual reconstruction - 1 case. In all cases the intra-surgical view motivated the operative intervention. The most important problem in treatment of the obstetric brachial plexus palsy is a separate group of children who need a surgical intervention. PMID- 16245209 TI - The lower extremity nerve injuries - own experience in surgical treatment. AB - The frequency of the lower extremity nerve injuries is assessed to about 20% of the overall lesions to the peripheral nerve system. Peroneal neuropathy is the most common lower extremity nerve palsy. In this study, results of the surgical treatment of the lower extremity nerve injuries have been presented. The clinical material consisted of 270 patients (192 males, 78 females aged from 3 months to 74 years) with injuries of the common peroneal nerve - 125, sciatic nerve - 93, common peroneal and tibial nerve - 21, tibial nerve - 17, femoral nerve - 10, others - 4. The following surgical procedures were performed: external neurolysis - 164, internal neurolysis - 27, reconstruction with sural nerve grafting - 63, direct neurorrhaphy - 12, neurotisation - 3, supplementary tenomioplasty - 23. Evaluation of the results with the use of BMRC scale (M0-M5) and Highet scale (S0 S4) included the group of 120 patients. After the surgical treatment a significant improvement was found in 63.3%. The efficacy of the treatment is strictly dependent on an early surgical intervention, mechanism and degree of the nerve injury as well as appropriate method of surgical therapy. PMID- 16245210 TI - CCL2 (MCP-1) and CCL5 (RANTES) levels in the peripheral blood of multiple sclerosis patients treated with Glatiramer Acetate (Copaxone). AB - The MCP-1 and RANTES levels were measured in 20 multiple sclerosis patients before and after 1 year daily treatment with 20 mg of subcutaneously applied glatiramer acetate. The level of MCP-1 in serum from multiple sclerosis patients was lower than in control subjects. After one year of therapy with glatiramer acetate, the level of MCP-1 was almost identical with that at the starting point. The concentration of RANTES in MS, both before and after therapy, did not differ from the control subjects. The results emphasise the marked difference between the influence of glatiramer acetate and IFNbeta-1a on the expression of the studied cytokine. Glatiramer acetate therapy in multiple sclerosis is not so much an effective as a protective factor of antiinflammatory cytokines, it should be regarded as a down-regulator of proinflammatory agents. PMID- 16245211 TI - Inhibition of poly(ADP-ribose) polymerase activity protects hippocampal cells against morphological and ultrastructural alteration evoked by ischemia reperfusion injury. AB - Poly(ADP-ribose) polymerase 1 (PARP-1 EC 2.4.2.30) is a nuclear enzyme that plays an important role in cell survival and death. PARP is involved in DNA repair machinery, however, massive DNA damage leads to over-activation of PARP-1 and to depletion of its substrate bNAD+ which causes cell death. Our previous study indicated that the PARP activity was significantly activated during ischemia reperfusion injury. In this study we investigated the effect of PARP inhibitor, 3 aminobenzamide (3-AB) on intracellular organelles alteration. Gerbils were submitted to 3 and 10 min transient global ischemia followed by recirculation and survival for 1 till 7 days. The histological and electron microscopic examination indicated a pronounced protective effect of 3-AB on the swelling of astrocytes and neurons 1 day after 3 and 10 min ischemic insult. It decreased also the swelling of pericytes. 3-AB decreases evoked by ischemia swelling of mitochondria and Golgi apparatus. The significant ameliorating effect of 3-AB was also observed on the 7th day of reperfusion after 3 min ischemia and was also visible on the 1st day after 10 min ischemia. However, 7 days after prolonged 10 min ischemia almost all neurons in the CA1 hippocampal layer died and 3-AB was not able to protect these cells. In spite of that, 3-AB markedly decreased immunostaining of glial fibrillary acidic protein (GFAP), which was enhanced in the stratum: oriens, radiatum and lacunosum-moleculare at the 7th day after 10 min ischemia. These data indicated that inhibition of PARP may have a protective effect on neuronal cells affected by ischemia-reperfusion injury. PMID- 16245212 TI - Ultrastructural study of cerebellar dentate nucleus astrocytes in chronic experimental model with valproate. AB - The current study focuses on the morphogenesis of changes in the cerebellum dentate nucleus in the course of experimental valproate encephalopathy. Valproate - a broad spectrum antiepileptic and antipsychotic drug - chronically used in rats, intragastrically, once daily at a dose of 200 mg/kg b. w. for 1, 3, 6, 9 and 12 months, induced pronounced ultrastructural changes in the population of glial cells and nerve cells of the dentate nucleus of the cerebellum in the last two phases of the experiment. Astrocytic and neuronal lesions coexisted with a considerable damage to the elements of the blood-brain barrier of the cerebellar structure examined. The changes affected mainly the population of protoplasmic astrocytes lying loosely in a neuropile as well as astrocytes adhering to damaged large multipolar neurons. Focal proliferation of astrocytes was observed. Abnormal astrocytes showed marked swelling expressed by significantly decreased electron density of the cytoplasm that contained almost empty vacuolar structures and by a considerably reduced number of intracellular organelles. It was accompanied by dilation of endoplasmic reticular channels, loss of fibrillopoietic capacity of the cell and features of autophagocytosis. It should be assumed that the essential cause of protoplasmic astroglial damage of the cerebellar dentate nucleus could be associated, apart from the direct effect of valproate and/or its metabolites on these cells, with changes in structural elements of the blood-brain barrier of this CNS region. PMID- 16245213 TI - Hemosiderin pigmentation of tumour cells in cerebellar pilocytic astrocytoma associated with post-traumatic hemorrhage in adults. AB - The pilocytic astrocytoma is only rarely associated with gross intratumoral hemorrhage despite rich vasculature and blood vessel changes, accompanied often by perivascular depots of hemosiderin. We report an unusual case of pigmented cerebellar pilocytic astrocytoma presenting with posttraumatic hemorrhage in a 38 year-old man with no history related to the tumor. CT and MRI examination after head injury demonstrated unexpectedly the cystic lesion of 2 cm in diameter in the region of the right cerebellar hemisphere and vermis. The lesion was associated with hematoma and it was surgically removed 3 weeks after trauma. Histopathological examination revealed pilocytic astrocytoma tissue with broad hemorrhagic changes and with an unusual pattern of massive pigmentation of the cytoplasm of pilocytic astrocytes, consistent with hemosiderosis. Positive stains for iron and ferritin and ultrastructural study confirmed deposition of hemosiderin granules in the tumour cells. There was no evidence of melanin or melanosomes. This finding of hemosiderin accumulation in the cytoplasm of neoplastic astroglia seems to be analogous to post-hemorrhagic pigmentation of the normal Bergmann glia and subpial astrocytes. In the literature, the examples of neuroepithelial tumors with hemosiderin pigmentation of tumor cells have been rarely documented. To our knowledge, this is the first reported case of pigmented pilocytic astrocytoma exhibiting extensive intracellular hemosiderin deposition. PMID- 16245214 TI - Meningeal solitary fibrous tumor: report of a case and literature review. AB - Solitary fibrous tumor is a rare neoplasm that most often involves the pleura. The increasing numbers of this neoplasm have also been reported to date in extrapleural sites. We report a case of a twenty-four-year-old female with right frontal mass. Histologically, the tumor composed of spindle cell proliferation. Tumor cells were found to be positive for CD34 and CD117 with immunohistochemical studies. Ten months follow-up was uneventful. Seventy seven cases of meningeal solitary fibrous tumor from the literature are analysed and pathological, immunohistochemical and clinical features are discussed. Solitary fibrous tumor has a slight female predominance, with a male to female ratio of 1:1.5. Age distribution is similar to meningioma ranging from 7-81 years. Approximately 23% of cases originate in the spine which is the most common meningeal location. Histopathologic examination shows uniform spindle cell proliferation with various amount of collagen. CD34-positivity usually allows discrimination from schwannomas, meningiomas and hemangiopericytomas. A differential diagnosis is important because most of the solitary fibrous tumors usually behave in a benign fashion. In this study, we also showed CD117 (Kit) expression in a case of meningeal SFT. CD117-positivity can be a good strategy for treatment in malignant and recurrent cases. Further investigations are necessary for therapeutic implication of CD117-positivity in SFT. PMID- 16245216 TI - Neuropathological diagnosis of tumour and bridge card game: some personal remarks and considerations (Letter to the Editor). PMID- 16245215 TI - Infantile mitochondrial leucodystrophy - a case report. AB - We retrospectively analyzed a case of a 7-month-old infant with a delay of psychomotor development, slow pupillary light reflexes, horizontal nystagmus, spasticity and bilateral optic nerve atrophy. At the end of life there were problems with swallowing. Ventriculography showed widening of the lateral ventricles and atrophy in the frontal lobes. EEG revealed generalized changes. Clinically, leucodystrophy was diagnosed. General autopsy revealed cardiac hypertrophy. Neuropathological picture showed orthochromatic leucodystrophy with some features characteristic of neuropathology of mitochondrial disease: capillary hyperplasia and hypertrophy, spongiosis and symmetrical, bilateral damage of brain stem structures. The last one is characteristic of Leigh syndrome. Electron microscopic evaluation showed abnormal mitochondria, myelin and neurofibrils destruction. Hypertrophy of the heart may be also connected with mitochondrial disease. PMID- 16245217 TI - The diagnosis and therapy of brain tumours. AB - Neoplasms arising from glial cells make up the most common group of primary brain tumors. The clinical outcome, especially the survival rates of the patients with brain tumours depend on tumour grade expressing its malignancy. A prognosis for glioblastomas (WHO IV) is very poor, but for astrocytomas (WHO I and II) it is relatively favourable. For oligodendrogliomas a longer survival time than for glioblastomas is observed. There is evidence that oxidative stress and reactive oxygen species (ROS) are crucial in the etiology and progression of a number of human diseases, including neoplasms. An oxidative damage of DNA, lipids and proteins is caused mainly with hydroxyl radical (*OH), the most reactive ROS species and may be seriously deleterious. In addition to all four basic nucleotides: adenosine (A), guanosine (G), tymidine (T) and cytosine (C), 5 methylcytosine (m5C) is a rare but normal component of cellular DNA and occurs mainly within a sequence of a structural gene or in regulatory regions. In the reaction with hydroxyl radical all DNA components can be modified, but m5C is relatively easily deaminated to thymine, which, in turn, pairs with adenine and after a round of replication, CG to TA transition occurs. Because thymine is a normal DNA base, therefore the product of spontaneous deamination of m5C is not so easily detected by cell's DNA repair system. Thus, 5-methylcytosine residue constitutes a mutational hotspot and DNA methylation pattern in patients might be useful as a primary diagnostic tool or as a marker for early detection of relapse of the disease. In recent years a new mechanism of posttranscriptional gene silencing has been discovered and named RNA interference (RNAi). This phenomenon is based on mRNA degradation mediated by small double-stranded RNA molecules, approximately 19-28 nucleotides in length, called short interfering RNAs (siRNAs). These molecules are produced from long dsRNAs by a dsRNA-specific endonuclease (DICER) and form 300 kD multi-enzyme complex (RISC) which by Watson Crick base-pairing of noncoding strand with their mRNA-targets induce specific degradation. The high sequence-specificity of RNAi makes it a new, promising tool in a gene-function analysis as well as in potential therapeutics development. PMID- 16245219 TI - A new marker for lipid peroxidation: serum paraoxonase activity in non-alcoholic steatohepatitis. AB - BACKGROUND/AIMS: Relationship between hepatic antioxidant paraoxonase 1 (PON1) activity, lipid peroxidation and liver injury was investigated in patients with non-alcoholic steatohepatitis. METHODS: A total of 23 patients with non-alcoholic steatohepatitis (15 males, 8 females; mean age: 40.30+/-7.67 yrs) and 23 healthy controls (14 males, 9 females; mean age: 39.70+/- 8.78 yrs) were enrolled in the study. Serum paraoxonase 1 activity and levels of a well-known lipid peroxidation marker, serum malondialdehyde, were determined. RESULTS: Serum paraoxonase 1 activity decreased significantly in non-alcoholic steatohepatitis compared to the control group (p<0.01). Serum malondialdehyde levels were significantly higher in patients with non-alcoholic steatohepatitis as compared with the control group (p<0.05). No statistically significant correlations were found between serum paraoxonase 1 activities and the grade-stage of non-alcoholic steatohepatitis, serum lipid levels or serum malondialdehyde levels (p>0.05). CONCLUSIONS: Increased lipid peroxidation may be either a cause or a result of liver injury in patients with non-alcoholic steatohepatitis. Although serum paraoxonase 1 activity does not reflect the degree of liver damage in non-alcoholic steatohepatitis, reduced paraoxonase 1 activity, especially in the presence of mild disease, could be interpreted as a biochemical marker of the lipid peroxidation. PMID- 16245220 TI - Management of fatty liver disease with vitamin E and C compared to ursodeoxycholic acid treatment. AB - BACKGROUND/AIMS: Despite a proposed role of oxidative stress in the pathogenesis of nonalcoholic steatohepatitis, antioxidant approaches have not been investigated sufficiently in the therapy of nonalcoholic steatohepatitis. Our aim was to determine whether vitamin E plus C therapy is effective in normalization of liver enzymes compared to ursodeoxycholic acid treatment in patients with fatty liver disease. METHODS: This was an open-labeled, prospective, randomized study enrolling patients with histologically proven fatty liver disease who had chronically elevated alanine aminotransferase, despite a three-month reducing diet. Patients consuming alcohol (more than 20 g/day) were excluded. The patients were randomly prescribed either oral vitamin E (600 IU/day) plus vitamin C (500 mg/day) or ursodeoxycholic acid (10 mg/kg/day). Patients were randomized as two groups to receive vitamin E plus vitamin C combination (28 patients, 10 F) or ursodeoxycholic acid treatment (29 patients, 13 F). RESULTS: There was no significant change in body mass index before and after the treatment in both groups. At the end of six months of therapy, serum aspartate aminotransferase and aminotransferase levels significantly decreased in both treatment options. Vitamin E and C combination was more efficacious on serum aminotransferase levels than ursodeoxycholic acid, but the difference was not significant. Alanine aminotransferase decreased to normal levels in 17 of 27 (63%) and in 16 of 29 patients (55%), respectively, in the two groups. Gamma-glutamyl transpeptidase decreased in patients receiving ursodeoxycholic acid, but no change was obtained in the vitamin-treated patients. CONCLUSIONS: Vitamin E plus C combination treatment is a safe, inexpensive and effective treatment option in patients with fatty liver disease, with results comparable to those obtained with ursodeoxycholic acid. Since more effective new therapeutic options are lacking, patients with fatty liver disease should be encouraged to take vitamin E and C supplements, which are safe and affordable. PMID- 16245221 TI - Importance of anticoagulant proteins in chronic liver diseases. AB - BACKGROUND/AIMS: This study was conducted to elucidate the importance of anticoagulant proteins in chronic liver disease and their possible role as markers in determining the severity of the liver disease. METHODS: This study was conducted on 35 patients with cirrhosis, 15 patients with chronic active hepatitis and 10 healthy controls. Coagulation inhibitor proteins such as protein C, protein S and antithrombin, as well as D-dimer level and thrombin time, which reflect fibrin degradation products, were measured. Cirrhotic patients were categorized as Child A, B and C and chronic active hepatitis patients as mild or moderate activity according to the modified Knodell histopathologic classification. The parameters were compared between patient groups and healthy controls. RESULTS: In comparison with controls, the cirrhotics had significantly decreased protein C, protein S, antithrombin levels and increased D-dimer levels. The Child B and Child C patients differed significantly with respect to protein C and antithrombin levels only. In the chronic active hepatitis patients, protein S, protein C and fibrinogen were within normal limits, whereas antithrombin was low. CONCLUSIONS: In chronic active hepatitis, the antithrombin level may be used as an early marker of hepatocellular damage. In cirrhotics, protein C and antithrombin may be useful for assessment of hepatocellular damage, whereas D dimer may be important for the transition to decompensation. PMID- 16245222 TI - Ultrasound-guided percutaneous cholecystostomy for acute cholecystitis in critically ill patients: one center's experience. AB - BACKGROUND/AIMS: The standard treatment for acute cholecystitis is cholecystectomy; however, cholecystectomy is not an option in some patients who are too high-risk for emergency surgery. Ultrasound-guided percutaneous cholecystostomy is an alternative for such patients. This study presents one center's five-year clinical experience with ultrasound-guided percutaneous cholecystostomy for treatment of acute cholecystitis. METHODS: In this study the records of all patients (18 total; mean age, 68.2+/-15.4 years; range, 42-91 years) who underwent ultrasound-guided percutaneous cholecystostomy for acute cholecystitis between June 1998 and October 2003 were reviewed. Duration of hospitalization, duration of tube placement, mortality and morbidity after tube placement, complication rates, culture results for aspirated bile, and clinical outcomes were analyzed. RESULTS: Fourteen patients were diagnosed with acute calculous cholecystitis and four were diagnosed with acalculous cholecystitis. The average hospital stay was 19+/-12.6 days (range, 5-52 days), and the average duration of catheter drainage was 20.5+/-19.1 days (range, 1-75 days). Six patients underwent open cholecystectomy between days 16 and 26 of catheter drainage, and none had postoperative complications. CONCLUSIONS: Ultrasound guided percutaneous cholecystostomy is a relatively safe and easy method for treating acute cholecystitis in critically ill patients. The risk of complications is low and the likelihood of success is high. PMID- 16245223 TI - The use and safety profile of non-steroidal antiinflammatory drugs among Turkish patients with osteoarthritis. AB - BACKGROUND/AIMS: To determine the use and safety profile of non-steroidal anti inflammatory drugs (NSAIDs) among Turkish osteoarthritis patients. METHODS: Osteoarthritis patients were interviewed by 138 doctors from clinics in nine different cities. Doctors completed a questionnaire regarding non-steroidal anti inflammatory drugs use and safety profile while interviewing the patients. RESULTS: Totally 3,755 patients (female/male: 3/1, mean age 59.0 +/- 12.2 years), 3,442 under non-steroidal anti-inflammatory drugs treatment, were included in the study. The use of meloxicam (5.5% vs. 14.4%) and specific cyclooxygenase-2 (COX 2) inhibitors (for celecoxib 3.3% vs. 12.2%; for rofecoxib 3.0% vs. 11.2%) increased more than that of other non-selective non-steroidal anti-inflammatory drugs. The most common side effects were epigastric burning (37%), other dyspeptic symptoms (25.3%), abdominal pain (17.0%), constipation (12.7%), nausea (10.6%) and diarrhea (3.0%). COX-2 selective and specific inhibitors had significantly lower incidence of dyspeptic complaints compared to non-selective non-steroidal anti-inflammatory drugs. No difference was found between the different non-steroidal anti-inflammatory drugs regarding the ratio of discontinuation of therapy due to inefficacy. The ratios of discontinuation due to side effects were lower in patients using COX-2 specific inhibitors compared to non-selective and selective non-steroidal anti-inflammatory drugs: celecoxib (7.7%), rofecoxib (10.3%), etodolac (12.4%), meloxicam (12.6%), tenoxicam (16.5%), diclofenac (16.8%), ibuprofen (19.4%), and naproxen (27.4%). Discontinuation of the non-steroidal anti-inflammatory drugs due to dyspeptic complaint was significantly less for specific COX-2 inhibitors than for non selective and selective non-steroidal anti-inflammatory drugs: celecoxib (2.5%), rofecoxib (8.4%), meloxicam (9.5%), etodolac (13.4%), tenoxicam (14.0%), diclofenac (14.1%), ibuprofen (17.2%), and naproxen (24.4%). CONCLUSIONS: The use of meloxicam and specific COX-2 inhibitors seems to have increased more than that of other non-selective non-steroidal anti-inflammatory drugs, if previously used non-steroidal anti-inflammatory drugs are considered. Fewer dyspeptic complaints have been reported with specific COX-2 inhibitors. PMID- 16245224 TI - Relationship between HLA-DR, HLA-DQ alleles and MEFV gene mutations in familial Mediterranean fever (FMF) patients. AB - BACKGROUND/AIMS: Three missense mutations clustered on the carboxyl-terminal portion of the MEFV gene (M680I, M694V, and V726A) have been observed in over 80% of affected alleles in several ethnic groups of familial Mediterranean fever patients. Several immunologic abnormalities were found both in cellular and humoral components in Mediterranean fever patients. Those observations have pointed the way for analysis of the HLA region in Mediterranean fever. We intended to compare HLA DR/DQ alleles with those major mutations in the MEFV gene in Mediterranean fever patients. METHODS: The distribution of MEFV gene mutations and HLA-DR, HLA-DQ alleles were analyzed in 40 index Turkish Mediterranean fever patients, 28 family members and 42 healthy controls. M680I, M694V, and V726A mutations were studied by amplification refractory mutation system-polymerase chain reaction (ARMS-PCR) analysis. HLA-DR and DQ allele subgroups were studied using SSP-PCR technique. RESULTS: A total of 37 (92.5%) patients in 40 Mediterranean fever index patients were found to carry one of the three missense mutations. The HLA-DR4 allele frequency was significantly higher in the Mediterranean fever patient group. When comparisons were made between Mediterranean fever mutations and HLA allele frequencies, M694V mutation with HLA DR3, DR11/5 and DR 13/6 and M680I mutation with DR7 allele subgroups were statistically significant. DQ6/1, DQ7/3, and DQ8/3 allele with M694V, DQ2 allele with M680I, and DQ6/1 with V726A mutations were also statistically significant. CONCLUSIONS: Our results indicate a relationship between some HLA-DR/DQ alleles and MEFV mutations in Mediterranean fever patients. We suggest HLA-DR/DQ alleles and their role in the pathogenesis of Mediterranean fever need further analysis and comparative studies. PMID- 16245225 TI - Can we rely on public data as a source of information for cancer registry in developing countries? AB - BACKGROUND/AIMS: Although a "hospital-based cancer registry" is important in improving patient care, a "population-based cancer registry" with emphasis on epidemiology is important in allocating health care resources and prioritizing public health programs. Because of its reliance on retrieved clinical and para clinical documents, there is some limitation in registering all cancer incidents in this system, especially in developing countries. In this study we examined the possibility of using public data as a complementary source of information for recording cancers in a population-based cancer registry. METHODS: Along with the annual census in rural areas, a survey was performed in Golestan province in March 2004 to identify public awareness about cancer incidents in the community. Individuals were questioned about history of cancer in their close relatives during the last two years. Those who reported cancer in their relatives were also asked to name the main organ of involvement. A similar list was retrieved from the cancer registry at the Ministry of Health in Gorgan, and cases with upper GI (esophagus and gastric) cancer diagnosis from 21 March 2002 through 20 March 2004 were selected for this study. Finally, these two lists were compared for examining accuracy of the collected data. RESULTS: We included 137 cases in our study with rural residence and known addresses. Only 35 (25.5%) cases were reported by the relatives and among them only 20 (57.1%) relatives correctly reported the tumor location. Although we found a difference in accurate reporting of cancer incidents by year of diagnosis (more correct cases reported during the second versus the first year), the difference was not statistically significant between the two years. CONCLUSION: In this study, we examined the possibility of using public awareness about cancer incidents as a complementary source of information for a population-based cancer registry. We found that this approach is not ideal for reducing limitations. Therefore, we recommend a nationwide cancer registry to record all cancer-related information at the time of diagnosis. This strategy will reduce the need for performing retrospective surveys in collecting cancer-related information. PMID- 16245226 TI - Gamna-Gandy bodies: a sign of portal hypertension. AB - Gamna-Gandy bodies (siderotic nodules) represent organized foci of hemorrhage in the spleen that is caused by portal hypertension. Gamna-Gandy bodies contain hemosiderin, fibrous tissue, and calcium. Magnetic resonance imaging has been approved as the most sensitive imaging modality for the detection of these nodules due to their iron content. Computerized tomography and ultrasonography also help in the detection and characterization of these lesions. We report here a case of portal hypertension due to cryptogenic liver cirrhosis with Gamna-Gandy bodies, and characteristic features of ultrasonography, computerized tomography and magnetic resonance. PMID- 16245228 TI - Pancreatic mass lesion mimicking carcinoma: initial presentation of retroperitoneal fibrosis. AB - A 63-year-old woman with upper abdominal pain radiating to her back was admitted to the hospital, and a mass lesion was identified involving pancreatic body and tail, with peripancreatic lymph node enlargement. The patient had normal tumor marker levels, a very high erythrocyte sedimentation rate (104) and negative malignant cells in fine needle aspiration cytology. The patient underwent laparotomy and histological findings of mass were consistent with retroperitoneal fibrosis. She has been in remission after long-term steroid treatment. In the differential diagnosis of cases with pancreatic mass lesion, retroperitoneal fibrosis should also be considered. PMID- 16245227 TI - Hepatocavopathy and isolated splenic vein thrombosis due to hypercoagulability state. AB - We report a patient with protein C and protein S deficiency and factor V Leiden mutation presenting with splenic vein thrombosis and with a web between the hepatic venous confluence and vena cava inferior. These two findings were thought to be due to the hypercoagulable state of the patient. Interestingly, there was no need for invasive procedures as the inferior accessory hepatic vein was patent. Hepatic venous flow was being maintained by the inferior accessory hepatic vein or a dominant collateral vein. PMID- 16245229 TI - Gastric heterotopia together with intestinal metaplasia in the gallbladder: case report and review of literature. AB - Heterotopic gastric mucosa in the gallbladder is extremely unusual. In this study, we aimed to report a case of gastric heterotopia together with intestinal metaplasia in the gallbladder of a 16-year-old male patient who experienced a sudden onset of epigastric pain with nausea. He was admitted to the hospital with a prediagnosis of mild degree obstructive jaundice. Cholecystectomy and hepaticoduodenostomy were carried out. In the microscopical examination of the gallbladder, an antral and pyloric type gastric mucosa together with intestinal metaplasia were clearly evident in the gallbladder submucosa, and the adjacent gallbladder mucosa showed typical features of chronic cholecystitis. PMID- 16245230 TI - Mitochondrial neurogastrointestinal encephalomyopathy. AB - Mitochondrial neurogastrointestinal encephalomyopathy is an autosomal recessive disease characterized by progressive ophthalmoplegia, peripheral neuropathy, mitochondrial abnormalities and gastrointestinal involvement. We describe a 19 year-old male having chronic intestinal pseudoobstruction associated with ophthalmoplegia and proximal muscle weakness. The clinical and radiologic features suggested the diagnosis of mitochondrial neurogastrointestinal encephalomyopathy. Mitochondrial genetic defects should be considered in the differential diagnosis of unexplained chronic gastrointestinal symptoms accompanied by neurological findings, especially in families where there is more than one individual with the same kind of symptoms. PMID- 16245231 TI - An uncommon lesion: gastric xanthelasma. AB - Although the clinical significance of gastric xanthelasmas is unclear, they are important lesions because they may be confused with malignant lesions. The etiopathogenesis is also unclear, but chronic gastritis, Helicobacter pylori (H. pylori) infection, diabetes mellitus and hyperlipidemia have been implicated. Xanthelasma is more frequent in women and its incidence increases with age. The lesions are frequently located in the stomach, and less frequently in the esophagus, duodenum and the colon. The lesions have a yellowish-white appearance, are between 0.5 and 10 mm in size and can be single or multiple. Xanthelasmas were found to be associated with chronic gastritis, gastrointestinal anastomoses, intestinal metaplasia, and H. pylori infection. These lesions are predisposing conditions for gastric cancer. Therefore, endoscopic biopsy is mandatory and careful follow-up is required. In this paper, four patients who attended hospital with abdominal pain and dyspepsia and by chance were found to have xanthelasmas on endoscopic examination are presented, and gastric xanthelasmas are discussed. PMID- 16245232 TI - Severe exudative ascites as an initial presentation of Crohn's disease. AB - Ascites in the course of Crohn's diseases is rarely seen and usually is a result of a concomitant malignancy, infection or thrombosis of portal or hepatic veins. In this article, a 55-year-old female patient suffering from severe ascites and finally diagnosed as Crohn's disease is presented. Further investigations revealed no malignancy, peritoneal infection or vein thrombosis. Treatment of Crohn's disease with steroids and 5-aminosalicylic acid also resulted in the disappearance of the ascites. PMID- 16245233 TI - Fulminant pseudomembranous colitis of the left colon successfully treated by surgical resection. AB - Pseudomembranous colitis is the classical and most dramatic manifestation of Clostridium difficile infection. Surgery is required for patients with perforation and for those who fail to respond to medical treatment. This is a report of a patient with severe pseudomembranous inflammation confined to the left colon, which was successfully treated by surgical resection. PMID- 16245234 TI - Upper gastrointestinal bleeding caused by simultaneous Dieulafoy's lesion and pre pyloric peptic ulceration. PMID- 16245235 TI - A patient with gastric carcinoid tumor: treatment and surveillance options. PMID- 16245236 TI - Histopathological atlas of renal diseases: anti-glomerular basement membrane antibody disease. AB - The main diagnostic feature of anti-glomerular basement membrane (anti-GBM) antibody disease is represented by the immunofluorescence pattern of intense and diffuse linear IgG deposition along the glomerular basement membrane. By light microscopy several histological patterns can be observed. PMID- 16245237 TI - Present and future therapy options in IgA-nephropathy. AB - Over the last 5 years the evidence base for therapeutic approaches in patients at risk for progressive IgA-nephropathy (IgAN) has markedly improved. In addition to several studies with low power, two randomized controlled trials suggest that in patients with preserved renal function, i.e. a GFR above 70 ml/min, corticosteroid-therapy can effectively prevent the development of progressive renal failure, whereas in patients with already impaired renal function, combination therapy consisting of cyclophosphamide, azathioprine and corticosteroids was effective. However, so far no study has firmly established that such immunosuppressive therapy is indeed superior to aggressive supportive therapy as it is available nowadays, i.e. high dose ACE-inhibitors and/or angiotensin-II receptor blockers, smoking cessation etc. It therefore appears prudent to first institute such supportive therapy and to limit immunosuppressive therapy to those patients, who maintain a proteinuria above 1 g/day or exhibit a progressive decline of GFR despite optimal supportive measures. Based on pathogenetic insights derived from experimental studies, antagonism of platelet derived growth factor-B or -D provides an attractive and possibly more specific therapeutical alternative to immunosuppression. While clinical studies testing this new approach are about to start, the development of other novel therapeutic approaches is greatly hampered by the lack of sufficient animal models of IgAN. In this respect a recently described primate model of IgAN offers some hope for the future. PMID- 16245238 TI - Polyunsaturated fatty acids and renal fibrosis: pathophysiologic link and potential clinical implications. AB - The role of polyunsaturated fatty acids in renal fibrosis. Several studies suggest a close relationship between polyunsaturated fatty acids (PUFA) and renal inflammation and fibrosis, which are crucial stages in chronic kidney disease (CKD). Beneficial effects of n-3 PUFA on the course of experimental and human nephropathies have been reported. PUFA can ameliorate chronic, progressive renal injury beyond the simple reduction of serum lipid levels. These pleiotropic effects of PUFA are due to their properties of interfering with the synthesis of a variety of inflammatory factors and events, through effects related both to the modulation of the balance of n-6 and n-3-derived eicosanoids and to direct action on the cellular production of the major cytokine mediators of inflammation and on endothelium function. The mechanisms by which PUFA can favorably interfere with some stages in renal fibrosis processes, such as mesangial cell activation and proliferation and extracellular matrix protein synthesis, include the regulation of some pro-inflammatory cytokine production, renin and nitric oxide (NO) systems and peroxisome proliferator-activated receptor gene expression. An optimal n-6/n 3 PUFA ratio dietary intake could offer new therapeutic strategies aimed at interrupting the irreversible process of renal fibrosis and ameliorating chronic renal injury. However, further experimental, epidemiological and clinical investigations are needed to confirm the role of PUFA in the renal fibrosis pathway and the natural history of chronic nephropathies. PMID- 16245239 TI - Chronic renal failure in male and female rats. AB - Gender influences the progression of chronic renal failure (CRF). We studied male (M) and female (F) Wistar rats for 90 days: castrated (CMc,n=7;CFc,n=6) and non castrated controls (CM,n=9;CF,n=6); castrated (CRFMc,n=8; CRFFc,n=6) and non castrated animals submitted to 5/6 nephrectomy (CRFM,n=13;CRFF,n=6). Data are expressed as mean +/-SEM. Proteinuria (PTN) was higher in CRFM (554+/-69 mg/24h) compared to CRFMc (277+/-85 mg/24h), but not in females (CRFF=193+/-20mg/24h, CRFFc= 164+/-71 mg/24h). Mesangial fractional volume increased in all CRF animals. CRF animals showed an increase of glomerular sclerosis index (GSI) and tubulointerstitial damage (TID) but in a smaller proportion in male castrated animals; the opposite occurred with females: castration induced an increase of these parameters. CRF animals showed increased cortical and glomerular fibronectin (FN) rates. Castration decreased glomerular and cortical FN rates in CRFM but not in females. In conclusion, proteinuria was higher in CRFM and probably led to glomerular and interstitial damage, as well as to FN accumulation, castration seems to protect against development of PTN, TID and FN accumulation in males. Castrated female rats presented mesangial expansion, with no changes in PTN, TID and FN rates. It seems that female sex hormones do not protect against renal disease progression, instead, we suggest that male sex hormones lead to acceleration of CRF. PMID- 16245240 TI - Endotoxin-induced acute renal failure in rats: effects of L-arginine and nitric oxide synthase inhibition on renal function. AB - BACKGROUND: The regulation of renal hemodynamics is closely related to the L arginine (L-Arg)/nitric oxide (NO) pathway. NO - metabolized from L-Arg - is capable of improving renal function in ischemic and toxic acute renal failure (ARF), while NO synthase (NOS) inhibition induces deterioration in renal function. The mortality rate in patients with septic shock is increased when treated with a non-selective NOS inhibitor, while the incidence of ARF requiring renal replacement therapy is unaffected. To date, there are no studies on the impact of NOS substrate (L-Arg) and inhibitor (L-NMMA) on renal function in early lipopolysaccharide (LPS)-induced ARF. METHODS: ARF was induced by intravenous (i.v.) LPS. Animals were treated with L-Arg, L-NMMA (NOS substrate and inhibitor), a combination of both or saline. Glomerular filtration rate (GFR), urine flow, fractional sodium excretion, excretion of NO metabolism stable end products and blood pressure (BP) were recorded at baseline, after ARF induction, during drug infusion and thereafter. RESULTS: L-Arg induced better GFR during infusion. Excretion of the NO metabolism end products was highest in the L-Arg group and lowest in the NOS inhibitor group. L-Arg administration had no influence on BP, while L-NMMA induced a slight elevation. CONCLUSIONS: We conclude that exogenous L-Arg exerts beneficial effects in early LPS-induced ARF in rats during drug infusion, while NOS inhibition has no influence on GFR. Subcellular compartmentalization of the L-Arg pool in cytoplasma and the rapid utilization of exogenous L-Arg in such a micro-environment could explain this effect, which has been observed in other ARF models and was called the "L-Arg paradox". In further studies the effects of early and prolonged administration of L-Arg in endotoxinemia should be investigated. PMID- 16245241 TI - Antibody-mediated pure red-cell aplasia (PRCA): the Spanish experience. AB - BACKGROUND: The incidence of antibody (Ab)-mediated pure red-cell aplasia (PRCA) in patients with chronic kidney disease (CKD) has increased between 1998 and 2002. After initially responding to treatment with recombinant human erythropoietic agents for CKD-associated anemia, patients became treatment refractory and severely anemic. Although most PRCA cases have occurred in Europe, the varying epidemiologies among individual countries have not been well characterized. METHODS: We investigated Ab-mediated PRCA in 12 Spanish patients treated with epoetin alfa alone or prior to treatment with epoetin beta (n=1) or darbepoetin alfa (n=1). Serum Abs against epoetin alfa were detected by radioimmunoprecipitation (RIP) assay or bioassay. Following diagnosis of PRCA, erythropoietic treatment was stopped and patients received immunosuppressive therapy alone (n=11) or in combination with renal transplant (n=1). RESULTS: Treatments were administered for 16 months (average) before diagnosis of PRCA in bone marrow aspirates (n=8) or biopsies (n=4). At diagnosis, patients had an average of 0.68% blood reticulocytes and blood hemoglobin (Hb) level of 7.13 g/dL. Eight patients had anti-epoetin Abs detected by RIP, and 5 had neutralizing Abs measured in the bioassay. As of December 2003, 4 patients had died, 3 had no recovery, and 5 had recovered from anemia (blood Hb level, 9.9 g/dL). All 5 recovering patients received corticosteroid therapy alone, and 1 received a renal transplant as well as corticosteroids. CONCLUSIONS: Sudden onset of treatment refractory anemia in CKD patients suggests a course of treatment cessation followed by diagnostic procedures for Ab-mediated PRCA, and immunosuppressive therapy. This study may serve as a model for a centralized global PRCA registry. PMID- 16245242 TI - Aortic pulse wave velocity and arterial wave reflections predict the extent and severity of coronary artery disease in chronic kidney disease patients. AB - BACKGROUND: Increased aortic stiffness markers - aortic pulse wave velocity (PWV) and augmentation index (AIx) - are powerful predictors of survival in ESRD patients - well-recognized for the high prevalence of coronary artery disease (CAD) and unusually high PWV and AIx. Recently, decreased aortic compliance has been shown to be predictive of primary coronary events in hypertensive patients with normal renal function. We aimed to explore relationships between arterial stiffness and CAD in cohorts of patients with chronic kidney disease (CKD). METHODS AND RESULTS: 46 patients with chronic kidney disease (33 males, aged 55.7+/- 13.2 years, 20 on dialysis, 18 post renal transplantation, and 8 with glomerular filtration rate (GFR) between 10 and 25 ml/min) underwent coronary angiography for the assessment of CAD. PWV and aortic AIx were determined from pulse waveform analysis of arterial waveforms recorded by applanation tonometry using a SphygmoCortm device. The atherosclerosis burden score was calculated by adding the percentage luminal reduction of the most severe lesion in each artery. Patients with normal angiograms had significantly less arterial stiffness (as reflected by both a lower PWV=8.42+/-1.53 m/s and a lower AIx=17.9+/-5.55 %) compared with the 35 subjects with evidence of obstructive coronary disease at angiography (PWV=9.21+/-1.15 m/s and AIx=23.4+/-5.4 %, P<0.05 for both). Moreover, as more coronary vessels were affected, PWV and AIx increased proportionally. Based on receiver operating characteristics (ROC) curve analysis mean PWV levels showed an optimal cut-off point at 8.35 m/s (sensitivity=0.77; specificity=0.60), while mean AIx levels showed an optimal cut-off point at 17% (sensitivity=0.87; specificity=0.70). There was a statistically significant linear relationship between the atherosclerosis burden and both measures of arterial stiffness: PWV (r=0.31, p=0.007) and AIx (r=0.46, p=0.003). Independent predictors for the arterial stiffness parameters in this CKD population (multiple stepwise regression analysis) were age (r=0.69 for PWV and r=0.62 for AIx), and mean arterial pressure (MAP) (for AIx, p<0.0001). CONCLUSION: This study provides the first direct evidence in a cross-sectional investigation that PWV and AIx are related to the extent of coronary obstruction in CKD patients. PMID- 16245243 TI - Management of hypertension in chronic kidney disease: the Italian multicentric study. AB - BACKGROUND: Guidelines have indicated the achievement of blood pressure target (BP <130/80 mmHg) as a priority in the conservative treatment of chronic kidney disease (CKD), but the current implementation of these recommendations in clinical practice is unknown. METHODS: We assessed control rates, treatment and clinical correlates of hypertension in 1201 adult non-dialyzed CKD patients followed up by a nephrologist for at least 6 months. RESULTS: Estimated glomerular filtration rate (GFR) was 32 (SD 15) mL/min/1.73 m2. BP target was not achieved in 88% of patients (95% confidence interval (95% CI): 86-90%). In 84% of patients, BP levels were also above the target at the first visit to the nephrology unit 4.5 yrs previously. The risk of not achieving BP target during the nephro-logy follow-up was associated with older age (odds ratio (OR): 1.24, 95% CI 1.06-1.45, p=0.008), diabetes (OR: 2.25, 95% CI 1.20-4.20, p=0.011), and the duration of hypertension (OR: 1.13, 95% CI 1.02-1.24, p=0.016). Among patients with uncontrolled BP, about 70% received multidrug antihypertensive therapy including renin-angiotensin system (RAS) inhibitors; conversely, diuretic treatment was prescribed in a minority of patients (37%), and at insufficient doses in half the cases, despite the insufficient implementation of a low salt diet (18%). CONCLUSIONS: BP target was not reached in most CKD patients routinely seen in the renal clinics. The main barrier to guideline implementation is possibly the inadequate treatment of extracellular volume expansion despite the large prevalence of factors, such as older age and diabetes, which further enhance the intrinsic BP salt sensitivity of CKD. PMID- 16245244 TI - Urinary GSTP1-1 excretion is markedly increased in normotensive pregnancy as well as in preeclampsia. AB - BACKGROUND: Glutathione S-transferases (GSTs) are present in large amounts in the human kidney, where they demonstrate a specific distribution. The assessment of urinary excretion of GST alpha (proximal tubules) and pi (distal and collecting tubules) could be helpful in determining if, and to what degree renal tubular damage is present in preeclampsia and whether this damage is in the proximal or distal region. METHODS: Urine samples were collected from 22 women with severe preeclampsia and/or HELLP syndrome (PE), from 30 non-pregnant women with a history of severe preeclampsia (HPE), from 18 women with uncomplicated pregnancies (PC) and from 30 non-pregnant women with a history of uncomplicated pregnancies (HPC). GSTA1-1 and GSTP1-1 were assayed by ELISA and were expressed as nanograms per 10 mmol creatinine (Cr). RESULTS: Median urinary GSTP1-1 concentrations were significantly (p<0.001) higher in women with preeclampsia [62.2 (4.3-291.2) ng/10 mmol Cr] compared to non-pregnant women with a history of preeclampsia [22.3 (0-142.6) ng/10 mmol Cr]). In addition, in normotensive pregnant women, urinary GSTP1-1 concentrations were significantly (p<0.01) higher [82.6 (8.3-206.7) ng/10 mmol Cr]) compared to non-pregnant controls [5.1 (0-66.7) ng/10 mmol Cr]. No difference in GSTP1-1 concentrations was found between women with preeclampsia and normotensive pregnant women. GSTA1-1 concentrations were not significantly different between the four groups of women investigated. There were no correlations between the degree of proteinuria and urinary GSTP1-1 or GSTA1-1 concentrations. CONCLUSION: GSTP1-1 metabolism in the distal tubule changes during normotensive as well as preeclamptic pregnancy. Whether this is due to tubular cell damage, disturbed resorption or an increase in cellular levels cannot be determined as yet. PMID- 16245245 TI - Predicting intradialytic hypotension from experience, statistical models and artificial neural networks. AB - BACKGROUND: Symptomatic intradialytic hypotension (IDH) associated with increased mortality in hemodialysis patients is difficult to predict and hence prevent. Artificial Neural Networks (ANNs) are promising tools to solve multidimensional non-linear problems. The aim of the study was to verify in which way mathematical models, statistics or knowledge of patients influence the ability of the nephrologists to predict IDH. METHODS: The performance of ANNs was compared with that of independent nephrologists supported by a logistic regression giving odds ratio for each studied variable (NEPHiS) or of nephrologists in charge of the patients without (NEPHc) or with statistical support as for NEPHiS (NEPHcS). Data from 98 hemodialysis patients were analysed in order to select patients with frequent IDH (>10% of the dialysis sessions). Complete data on 1979 dialysis sessions from 7 patients were retrieved. The ability to predict the occurrence of hypotension episodes was compared (ROC curves) between ANNs, NEPHc/S (N=7) in Switzerland and NEPHiS from independent dialysis centers in Western Australia (N=10). RESULTS: ANN gave the most accurate correlation between estimated and observed IHD episodes compared to NEPHc (p<0.001), but a similar performance was attained by NEPHcS (p<0.001). NEPHiS were superior to NEPHc (P<0.05), but inferior to ANN (P<0.01). For a sensitivity of 80%, specificity was 44% for ANNs, 33% for NEPHcS and 20% for NEPHc. CONCLUSIONS: ANNs are superior to nephrologists in predicting IDH episodes; however when supported by a statistical analysis, nephrologists reach ANNs in their prediction ability. IDH still remains difficult to predict even with mathematical models. PMID- 16245246 TI - Lack of significant circadian and post-prandial variation in phosphate levels in subjects receiving chronic hemodialysis therapy. AB - BACKGROUND: The present study was designed to determine the reliability of the current practice of random pre-dialysis phosphate testing in subjects receiving hemodialysis therapy, since phosphate levels exhibit a significant variation in a 24-hour period under usual physiologic conditions. METHODS: Subjects receiving chronic hemodialysis (HD) were invited to participate during an incidental hospitalization. In Study A subjects (n=31) had serum phosphate tested three times on a single non-dialysis day, between 6 to 7 am (A1), between 11 am to 12:30 pm (A2), and 3:45 pm to 4:45 pm (A3). In study B subjects (n=25) had serum phosphate tested just before (B1) and 2 hours after lunch (B2), on a non-dialysis day. For Study A the collection times coincided with the start times of the out patient dialysis shifts at our institution. All patients continued their usual phosphate binder therapy, if any. For study A the results were analyzed using one way repeated measures analysis of variance or Friedman repeated measures analysis of variance on ranks, as opposite. Paired t-test was used for Study B. Results are expressed as mean+/- standard deviation. RESULTS: Twenty-three men and eight women (mean age 62.2+/-15.2 years) were enrolled in Study A (24 Caucasian, 7 African-American). Nineteen men and six women (mean age 65.5+/-12.8 years) participated in study B (19 Caucasian, 6 African-American). In Study A, there was no significant difference in the mean serum phosphate levels in the samples collected through the day A1 (4.5+/-1.3 mg/dL), A2 (4.5+/-1.3 mg/dL) and A3 (4.7+/-1.5 mg/dL) (p=0.19 for comparison of the three). The mean amplitude of circadian variation (peak minus trough) was 0.64+/- 0.37 mg/dL. Similarly, there was no significant difference in the mean serum phosphate before (4.4+/-1.4 mg/dL) and two hours after lunch (4.4+/-1.5 mg/dL) (p =0.6). The mean of the difference in serum phosphate (post-prandial minus pre-prandial) was 0.2+/-0.4 mg/dL. CONCLUSION: Our results in hospitalized HD subjects indicate there is no significant difference in phosphate levels at different times of a single day or in relation to meals. PMID- 16245248 TI - Mycophenolate mofetil in the treatment of steroid-resistant primary focal segmental glomerulosclerosis. AB - We report our experience in using mycophenolate mofetil (MMF) for the treatment of steroid-resistant focal segmental glomerulosclerosis (FSGS) in two patients. Patient 1, who was treated on disease presentation, responded well with sustained complete remission. Patient 2, who had unsuccessful steroid treatment 4 yrs before and was re-treated with MMF, showed transient retardation in renal disease progression but eventually progressed to end-stage renal failure. Our observation illustrates that MMF could be useful in treating steroid-resistant FSGS if administered at an early phase of the disease, well before histologic damage becomes irreversible. Its efficacy requires validation in randomized, controlled trials. The current armamentaria for the treatment of this condition are also discussed. PMID- 16245247 TI - Atherosclerotic renovascular disease: medical therapy versus medical therapy plus renal artery stenting in preventing renal failure progression: the rationale and study design of a prospective, multicenter and randomized trial (NITER). AB - BACKGROUND: Many studies suggest a major prevalence of atherosclerotic renovascular disease (ARVD), caused by mono or bilateral renal artery stenosis (RAS). Unfortunately, there is no definite therapy to cure this disease to date; therefore, ARVD is burdened by important clinical complications with high social and economic costs. The last few years have seen important advancements in both medical therapy and in interventional radiology (for example, percutaneous transluminal renal artery stenting (PTRS)). All of them could affect, in some way, the natural history of ARVD, but to date the optimal strategy has not been established. METHODS: The protocol of a prospective, multicenter, randomized trial "Nephropathy Ischemic Therapy (NITER)" is presented. It enrolls patients with stable renal failure (glomerular filtration rate (GFR) >or=30 ml/min) and hypertension, and hemodynamically significant atherosclerotic ostial RAS (>or=70%) diagnosed by duplex Doppler (DD) ultrasonography and confirmed by magnetic resonance angiography (MRA). This study aims to evaluate whether medical therapy plus interventional PTRS is superior to medical therapy alone according to the following combined primary endpoint: death or dialysis initiation or reduction by >20% in estimated GFR after 0.5, 1, and 2 yrs of follow-up and an extended follow-up until the 4th year. Medical therapy means drugs to control hypertension, improve dyslipidemia and optimize platelet anti-aggregant therapy. The sample size is estimated in 50 patients per group to achieve a statistical significance of 0.05 in case of a reduction by 50% in the combined endpoints. PMID- 16245249 TI - Hepatitis B complicated focal segmental glomerulosclerosis. AB - After the initial report of membranous glomerulopathy due to hepatitis B virus infection by Combes et al, other glomerular diseases - but rarely focal segmental glomerulosclerosis (FSGS) association with HBV infection - have been reported. Herein we present an 8-year-old boy with chronic HBV infection complicated FSGS. The patient was initially regarded as idiopathic FSGS and started on an immunosuppressive schedule. The elevation of liver transaminases in the course of the therapy revealed the immunotolerated perinatal HBV infection. It was considered that immunosuppressive agents have induced viral replication. The treatment was changed to lamivudine alone. The nephrotic syndrome has already been improved with the seroconversion of anti-HBeAg and reduced liver functional tests by the tenth month of the treatment. This case is peculiar for the seldom association of FSGS with chronic HBV infection and treatment modality particularly for the countries where this viral infection is endemic. PMID- 16245250 TI - Crescentic glomerulonephritis associated with renal cell carcinoma after cancer immunotherapy. AB - A 59 year-old woman showed rapidly progressive glomerulonephritis during immunotherapy for metastatic renal cell carcinoma. She received unilateral nephrectomy and cytotoxic T lymphocyte (CTL) therapy for the treatment of retroperitoneal lymph node metastasis of renal cell carcinoma. With CTL therapy, her retroperitoneal lymph node mass decreased in size. One year after the third round of CTL therapy, her serum creatinine was increased and massive proteinuria occurred. Her renal biopsy specimen revealed necrotizing and crescentic glomerulonephritis with immune complex deposition. Her retroperitoneal lymph node mass continued to decrease in size. Consequently, for the purpose of avoiding interfering with the CTL therapy, we performed double filtration plasmapheresis (DFPP) monotherapy for removal of immune complexes without using immunosuppressive drugs or prednisolone. After 24 sessions of DFPP, her serum IgG was reduced from 3,942 mg/dL to 2,400 mg/dL, and proteinuria (from 9.0 g/day to 0.9 g/day) and renal function (serum creatinine; from 5.6 mg/dL to 2.2 mg/dL) also improved. However, 3 months after the final DFPP, she expired due to perforation of the colon. The autopsy sample of the kidney showed that most of the glomeruli were obsolescent, but immunoglobulin depositions were reduced and necrotizing lesions were diminished. In the patients with RPGN associated with renal cell carcinoma, renal functional recovery has not been observed upon immunosuppressive treatment. Consequently, plasmapheresis is considered to be one of the effective and safe methods for patients with this association. We also discuss previous reports of RPGN associated with renal cell carcinoma, or RPGN after cancer immunotherapy. PMID- 16245251 TI - Pseudo-renal failure following total abdominal hysterectomy. AB - Intraperitoneal urinary bladder perforation should be in the differential diagnosis of acute oliguric renal failure soon after gynecological surgery. We present a case of reversible acute pseudo-renal failure after total abdominal hysterectomy for uterine fibroid. Biochemical features of uremia occur as a result of intraperitoneal extravasation of urine, which is in turn reabsorbed through the peritoneum. Early recognition and surgical repair, as opposed to dialysis therapy, are warranted in such clinical setting. Nephrologists, who are often the first to encounter those patients with presumably acute renal failure, should be aware of this condition. Prompt recovery of the serum biochemistry is to be expected, in contradistinction to genuine renal failure or kidney insults. PMID- 16245252 TI - HPRT deficiency as the cause of ESRD in a 24-year-old patient: a very rare presentation of the disorder. AB - A 24-year-old male with end-stage renal disease (ESRD) and disproportionately high uric acid plasma concentration was admitted to our unit. After studying the patient's medical history, as well as that of the entire family, hyperuricemia was discovered in his brother, while microscopic examination of his brother's and mother's urine revealed abundant uric acid crystals. After performing purine metabolic studies, it was determined that the two siblings suffered from partial hypoxanthine-guanine phosphoribosyltransferase (HPRT) deficiency (Kelley Seegmiller syndrome). This report highlights the importance of clinical awareness and a thorough examination of the patient's medical history for establishing an early diagnosis and commencing treatment for such rare inherited metabolic disorders to prevent renal failure. PMID- 16245254 TI - Oxidative and nitrosative stress in kidney disease: a case for cyclosporine A. AB - The immunosuppresor cyclosporine A (CsA) has been associated to human endothelial dysfunction and accelerated atherosclerosis. Sympathetic overactivity, relative deficiency of nitric oxide, TGFb-1, endothelin-1, reactive oxygen (ROS) and nitrogen species (RNS) and vasoconstrictor eicosanoids are mediators of vascular dysfunction associated to cyclosporine A. In CsA-treated cells (BAEC) an increase in reactive oxygen and nitrogen intermediates may lead to the intracellular formation of peroxynitrite. This agent could be one important mediator by which CsA produces an antioxidant-sensitive nitration of tyrosine, a marker for endothelial damage by nitrosative stress. Superoxide anion is the limiting factor in the formation of peroxynitrite in CsA-treated endothelial cells. Treatment with CsA may lead to the nitration of specific proteins such as manganese superoxide dismutase (MnSOD). We propose that peroxynitrite and tyrosine nitration may represent mechanisms of damage in pathophysiological situations where superoxide anion generation is increased. PMID- 16245255 TI - Bone metabolism, vascular calcifications and mortality: associations beyond mere coincidence. AB - Bone and cardiovascular disorders are common age-related disorders in the general population and also in patients suffering from chronic kidney disease (CKD). Recent studies have shown an association between these two disorders and the rate of mortality. This article addresses some limitations of the concept of osteoporosis in CKD and compares bone and vascular disorders and mortality between non-selected general population and dialysis patients from the same geographic area. In the general population, all metabolic disorders increase with age, as well as vascular calcifications. The progression of vascular calcification was associated with a higher prevalence and incidence of bone fractures. In addition, both vascular calcifications and vertebral fractures were associated with higher mortality. A similar pattern was observed in dialysis patients with no increments in vertebral fractures, although with higher prevalence of vascular calcifications also both associated with higher mortality. Age was the strongest variable associated with all the analysed parameters, but some of the associations remained significant after age adjustment indicating the likely role of other common factors in the pathogenesis of bone and vascular disorders. PMID- 16245256 TI - Leptin in end stage renal disease (ESRD): a link between fat mass, bone and the cardiovascular system. AB - Adipose tissue is now considered an important system operating strictly in concert with other systems. The adipocyte is the main producer of two pleiotropic compounds, leptin and adiponectin, modulating inflammation and having multiple effects in disparate organs including the cardiovascular and the central nervous system. Leptin has disparate influences on various physiologic and organ systems including glucose homeostasis, hematopoiesis and the reproductive and cardiovascular systems and is a crucial hormone for the regulation of food intake and body weight. Peripherally, leptin modulates insulin sensitivity and high leptin triggers insulin resistance and vice versa. Obesity, a situation where circulating leptin attains very high levels is accompanied by increased bone mass, a phenomenon which may depend on direct stimulation of osteoblasts by leptin. However in animal models the stimulating effect of leptin on the osteoblast is counterbalanced by a strong inhibitor effect on bone formation in the central nervous system. Two recent studies reported an inverse link between leptin, bone mass and PTH in dialysis patients suggesting that leptin may be implicated in low bone turnover in these patients, likely by a mechanism involving the central nervous system. Leptin induces vascular calcifications in vitro. In uremic man leptin is unrelated to valvular calcifications but predicts incident cardiovascular events in overweight and obese dialysis patients. Leptin seems to be a relevant player in the emerging connection between bone and cardiovascular alterations in patients with end stage renal disease. PMID- 16245257 TI - Genetics and molecular disorders in severe secondary hyperparathyroidism: lessons from rna and microarray studies. AB - Secondary hyperparathyroidism-related disorders begin in the pre-dialysis period and progressively worsen during dialysis. In a high proportion of cases, therapeutic failure in the control of PTH secretion is related to a late start in medical treatment. This may happen because recovery of the functional control of the parathyroid gland, once some irreversible molecular and genetic changes have occurred, can be only partial. These irreversible changes include promotion of cell proliferation and failure of several pathways affecting the metabolism of DNA, RNA and proteins. PMID- 16245258 TI - Acute pyelonephritis: a new approach to an old clinical entity. PMID- 16245259 TI - Sequence diversity of the peptaibol antibiotic suzukacillin-A from the mold Trichoderma viride. AB - From the culture broth of the mold Trichoderma viride, strain 63 C-I, the polypeptide antibiotic suzukacillin (SZ) was isolated. A peptide mixture named SZ A was obtained by crystallization from crude SZ. Individual peptides from SZ-A were isolated by semipreparative HPLC and sequences were determined by HPLC-ESI MS. The data confirm a general sequence of SZ-A published previously and in addition establish the individual sequences of 15 acetylated eicosa peptides with C-terminal alcohols. The major peptide SZ-A4 (21% of all peptides) shows the sequence:Ac-Aib-Ala-Aib-Ala-Aib-Ala(6)-Gln-Aib-Lx(9)-Aib-Gly-Aib(12)-Aib-Pro Vx(15)-Aib-Vx(17)-Gln-Gln-Fol. Amino acid exchanges of the peptaibol are located in position 6 (Ala/Aib), 9 (Vx/Lx), 12 (Aib/Lx), 17 (Aib/Vx) and possibly at position15 (Val/Iva) (uncommon abbreviations: Aib (alpha-aminoisobutyric acid); Iva (D-isovaline); Lx (L-leucine or L-isoleucine); Vx (L-valine or D-isovaline); Fol (L-phenylalaninol)). PMID- 16245260 TI - Solid-supported and pegylated H-Pro-Pro-Asp-NHR as catalysts for asymmetric aldol reactions. AB - H-Pro-Pro-Asp-NH2 is a highly active and selective catalyst for asymmetric aldol reactions. Here, the versatility of H-Pro-Pro-Asp-NH2 has been further improved by immobilization on a solid support and functionalization with a short polyethylene glycol linker at the C-terminus. The development, synthesis, and the catalytic properties in aldol reactions of H-Pro-Pro-Asp-resin and H-Pro-Pro-Asp Ahx-NH(CH2CH2O)3CH3 are described. For the solid-supported catalyst, TentaGel with a loading of 0.1-0.2 mmol g(-1) proved to be the optimal support. The solid supported catalyst can be recycled at least three times without a significant drop in the catalytic activity or selectivity. Using the pegylated catalyst H-Pro Pro-Asp-Ahx-NH(CH2CH2O)3CH3, only 0.5 mol % are necessary to obtain aldol products in up to 96% yield and 91% enantiomeric excess. In all cases, enantioselectivities are comparable to those obtained with the parent catalyst H Pro-Pro-Asp-NH2. Thus, immobilization of H-Pro-Pro-Asp-NH2 on Tentagel as well as pegylation led to catalysts with selectivities comparable to the nonmodified catalyst, exhibiting additional distinct advantages such as facile reusability, ease of handling, higher solubility, and thereby greater versatility. handling, higher solubility, and thereby greater versatility. PMID- 16245261 TI - Structure optimization in a three-dimensional off-lattice protein model. AB - We studied a three-dimensional off-lattice AB model with two species of monomers, hydrophobic (A) and hydrophilic (B), and present two optimization algorithms: face-centered-cubic (FCC)-lattice pruned-enriched-Rosenbluth method (PERM) and subsequent conjugate gradient (PERM++) minimization and heuristic conjugate gradient (HCG) simulation based on "off-trap" strategy. In PERM++, we apply the PERM to the FCC-lattice to produce the initial conformation, and conjugate gradient minimization is then used to reach the minimum energy state. Both algorithms have been tested in the three-dimensional AB model for all sequences with lengths 13 < or = n < or = 55. The numerical results show that the proposed methods are very promising for finding the ground states of proteins. In several cases, we renew the putative ground states energy values. PMID- 16245262 TI - Simple and versatile restraints for the accurate modeling of alpha-helical coiled coil structures of multiple strandedness, orientation and composition. AB - We present a minimalist approach for the modeling of the three-dimensional structure of multistranded alpha-helical coiled coils. The approach is based on empirical principles introduced by F. H. C. Crick (F. H. C. Crick, Acta Crystallogr, 1953, Vol. 6, pp. 689-697). Crick hypothesized that keeping the distance between the residues at the interacting interface of alpha-helices constant would lead to supercoiling or the formation of a coiled coil through the knobs-into-holes mode of packing. We have implemented the latter hypothesis in a simulating annealing protocol in the simple form of interhelical distance restraints (two per heptad) between Calpha at the interfacial positions and. To demonstrate the authenticity of Crick's hypothesis and the precision and accuracy of our approach, we have modeled the crystal structures of six synthetic coiled coils in dimeric, trimeric, and tetrameric states. The mean root mean square deviations (RMSDs) between the backbone atoms of the ensemble of structures calculated and those of the corresponding geometric averages is always below 0.76 A, indicating that our protocol has an excellent degree of convergence and precision. The RMSDs between the backbone atoms of each of the six geometric average structures and the backbone of the corresponding crystal structures all range between 0.43 and 0.95 A, indicative of excellent accuracy and proving the authenticity of Crick's hypothesis. Moreover, without specifying any dihedral angles, we found that in 81% of the occurrences, the most populated conformer of the side chains at positions and in the ensembles calculated were identical to those observed in the crystal structures. This shows that our simple approach, which is the simplest reported so far, can generate accurate results for the backbone and side chains. Finally, as a test case for a wider application of our approach in the field of structural proteomics, we describe the successful modeling of the overall structure of SNARE and the organization of its interfacial ionic layer known to play an important functional role. PMID- 16245263 TI - Clinical application of an enzyme immunoassay for cholecystokinin-like immunoreactive substance for determination of the human plasma levels: the effect of metoclopramide on gastrointestinal peptides and stress-related hormones. AB - Metoclopramide, a prokinetic drug, is widely used to treat vomiting and nausea. Delayed gastric emptying and continual stress are considered important factors, among others, that induce nausea and vomiting. One gastrointestinal motility regulatory factor has been assumed to be the induction of changes in the levels of peptides such as gastrin, somatostatin, motilin, and cholecystokinin (CCK) in plasma. In contrast, adrenocorticotropic hormone (ACTH) and cortisol are used as indicators of stress. Here, we studied the effects of metoclopramide on human plasma gastrin-, somatostatin-, motilin-, and CCK-like immunoreactive substances (ISs) and ACTH-IS and cortisol under stress conditions using repetitive blood sampling in healthy subjects. Metoclopramide hydrochloride at a dose of 30 mg or placebo was orally administered to five healthy male volunteers. Blood samples were taken before and 20, 40, 60, 90, 120, 180, and 240 min after administration, subject to extracting procedures, and submitted to a highly sensitive enzyme immunoassay system. A single administration of metoclopramide caused significant increases in plasma somatostatin-IS levels compared with the placebo. Metoclopramide significantly decreased plasma gastrin- and suppressed ACTH-IS and cortisol levels compared with the placebo. We hypothesize that metoclopramide might have an accelerating gastric-emptying effect and a modulatory effect on the hypothalamo-pituitary-adrenal (HPA) axis and the autonomic nervous function. These effects might be beneficial in stress-related diseases, which suggest that this medicine has clinicopharmacological activities. PMID- 16245265 TI - A powerful Bronsted acid catalyst for the organocatalytic asymmetric transfer hydrogenation of imines. PMID- 16245264 TI - Synthesis of oligotuftsin-based branched oligopeptide conjugates for chemotactic drug targeting. AB - The synthesis and chemotactic properties of a new class of branched oligopeptide based conjugates are described. Tetratuftsin derivatives containing chemotactic formyl tripeptides (For-MLF, For-NleLF or For-MMM) in branches were prepared by stepwise solid-phase peptide synthesis. The influence of the composition and ionic charge of the carrier-branched oligopeptide on the chemotactic behaviour of the conjugate was studied in Tetrahymena pyriformis. Conjugates with methotrexate (Mtx) as a drug component was also prepared. For this, a GFLGC spacer, cleavable by cathepsin B, was used. The spacer with N-terminal methotrexate was coupled to the chloroacetylated chemotactic carrier molecule by thioether bond formation. The chemotactic activity and cytotoxity of Mtx conjugates were also studied. PMID- 16245266 TI - Biocomposites of plasticized starch reinforced with cellulose crystallites from cottonseed linter. AB - Environmentally friendly starch biocomposites were successfully developed using a colloidal suspension of cottonseed linter cellulose crystallite as a filler to reinforce glycerol plasticized starch (PS). The cellulose crystallites, having lengths of 350 +/- 70 nm and diameters of 40 +/- 8 nm on average, were prepared from cottonseed linters by acid hydrolysis. The dependence of morphology and properties of the PS-based biocomposites on cellulose crystallites content in the range from 0 to 30 wt.-% was investigated by scanning electron microscopy, differential scanning thermal analysis, dynamic mechanical thermal analysis, and measurements of mechanical properties and water absorption. The results indicate that the strong interactions between fillers and between the filler and PS matrix play a key role in reinforcing the resulting composites. The PS/cellulose crystallite composites, conditioned at 50% relative humidity, undergo an increase in both tensile strength and Young's modulus from 2.5 MPa for PS film to 7.8 MPa and from 36 MPa for PS film to 301 MPa. Further, incorporating cottonseed linter cellulose crystallites into PS matrix leads to an improvement in water resistance for the resulting biocomposites. The mechanical behaviors of the starch-based biocomposites as a function of cellulose crystallites content. PMID- 16245267 TI - Intrinsic adhesion properties of poly(vinyl pyrrolidone) to pharmaceutical materials: humidity effect. AB - Poly(vinyl pyrrolidone) (PVP) is widely used for bioengineering and pharmaceutical applications, and its adhesion characteristics are critical. When used as a binder in pharmaceutical granulation, it covers the resultant granules and governs their surface properties. The intrinsic adhesion forces of PVP toward common hydrophobic (magnesium stearate) and hydrophilic (lactose) pharmaceutical materials have been studied as a function of relative humidity (RH). The effect of RH on adhesion force was more significant for the PVP/hydrophilic material than the PVP/hydrophobic material. Adhesion was lowest between 20 and 40% RH, and it increased at RH above 40% and below 20%. This is likely to be due to the development of capillary and triboelectrification forces, respectively. In a nano indentation experiment using a silicon tip at room temperature, the PVP surface underwent a glass transition at 70% RH. This result suggests that surface softening contributes to the increased PVP adhesion at RH above 70%. To adjust the adhesion properties of PVP, humidity control should be an essential part of research and development. Effect of humidity on the adhesion forces between PVP and lactose (LT) or magnesium stearate (MS). PMID- 16245268 TI - A novel injectable poly(epsilon-caprolactone)/calcium sulfate system for bone regeneration: synthesis and characterization. AB - A novel poly(epsilon-caprolactone)/calcium sulfate system was prepared and characterized in order to enhance calcium sulfate (gypsum) performance as bone graft substitute overcoming its brittleness and fast resorption rate. A poly(epsilon-caprolactone) (PCL) photo-crosslinkable derivative (PCLf) was synthesized by reaction of a low molecular weight PCL diol with methacryloyl chloride and confirmed by FT-IR and 1H NMR analyses. An injectable and easy mouldable mixture of PCLf and calcium sulfate hemi-hydrate (PCLf/CHS) was obtained. Thermal analyses and solvent extraction proved the occurrence of PCLf photo-crosslinking, even in the presence of CHS, in a time suitable for clinical applications. Swelling studies demonstrated that the encapsulation of the inorganic filler increases network hydrophilicity making it more permeable to water. Scanning electron microscopy, performed on crosslinked PCLf/CHS and on the same material after incubation in a PBS solution, showed the feasibility to obtain, in situ, gypsum entrapped into a degradable polymeric network. In vitro cytotoxicity tests, performed according to ISO 10993-5, proved that the developed system was not cytotoxic supporting its potential use in tissue engineering as a new, injectable, photocurable bone graft material. SEM micrograph of calcium sulfate di-hydrate (gypsum) entrapped in the PCL network. PMID- 16245269 TI - pH-induced micelle formation of poly(histidine-co-phenylalanine)-block poly(ethylene glycol) in aqueous media. AB - Poly[(L-histidine)-co-(L-phenylalanine)]-block-poly(ethylene glycol) (HF-b-PEG) diblock copolymers were synthesized to be used for preparation of pH-sensitive polymeric micelles. First, HF block was synthesized by ring opening copolymerization of L-histidine and L-phenylalanine N-carboxyanhydride, and then the resulting copolymer was coupled with PEG. The pKa value of diblock copolymer can be controlled by adjusting the histidine/phenylalanine ratio in HF block. It is observed that the block copolymers form micelles in aqueous media and that the micelles are spherical in shape with a unimodal distribution. The micelle is formed at pH higher than pKa of block copolymer while it is not formed at lower pH. This is because the protonation of histidine residue in the HF block converts the hydrophobic core into hydrophilic one at lower pH. Acid-Base titration profile of HF41(5600)-b-PEG, HF56(5500)-b-PEG, H(5100)-b-PEG and 0.1 N NaCl. PMID- 16245270 TI - Synthesis and antioxidant efficiency of a new copolymer containing phosphorylated myo-inositol. AB - New data are constantly gathered to show the role of oxidative stress and the involvement of reactive oxygen species in the pathogenesis of degenerative diseases. InsP6 is able to coordinate iron metal in order to prevent iron catalyzed free radical formation. The aim of the present paper is to describe a new synthetic strategy in order to prepare a polymeric structure containing chemical functions able to coordinate iron ions. Here, we report the synthesis of a copolymer containing phosphorylated myo-inositol groups and we evaluate its antioxidant efficiency. Such a system was synthesized by binding chemical groups susceptible of radical polymerization to myo-inositol. The synthesized monomer was copolymerized with N,N-dimethylacrylamide (DMAA) (molar ratio 1:3) and submitted to exhaustive phosphorylation. The reaction was proved by an assay specific for phosphate groups. Finally, we evaluated the copolymer's ability in inhibiting lipid peroxidation in rat liver microsomal membranes. This study showed that the designed macromolecular system is particularly effective as antioxidant. PMID- 16245271 TI - Ternary complex formation in aqueous solution between a beta-cyclodextrin polymer, a cationic surfactant and DNA. AB - Polyelectrolyte complexes have been elaborated by mixing in water neutral poly(beta-CD), a cationic surfactant (DTAC) and herring sperm DNA fragments. The driving forces for the poly(beta-CD)/DTAC/DNA association in aqueous solution are, on the one hand, reversible inclusion interactions between the CD cavities of poly(beta-CD) and the alkyl group of DTAC, leading to the formation of a polycation and, on the other hand, electrostatic interactions between the opposite charges of the cationic surfactant and anionic DNA. Viscometry and SANS have been used to prove the occurrence of such ternary complexes in dilute aqueous solutions. PMID- 16245272 TI - Preparation and interfacial properties of a novel biodegradable polymer surfactant: poly(ethylene oxide monooleate-block-DL-lactide). AB - In this paper, we report a novel synthesis of poly(ethylene oxide monooleate block-DL-lactide) (MOPEO-PLA) in the presence of stannous 2-ethylhexanoate catalyst. By utilizing the surfactant property and the reactive double bond of the amphiphilic MOPEO-PLA, various characteristics of PLA microspheres, such as surface and internal structure, surface morphology, release property, and so on, may potentially be controlled. MOPEO-PLA was found to be hydrophobic enough to prevent loss by dissolution into aqueous solution, which is often a problem for MOPEO. Furthermore, the interfacial tension measurements of a MOPEO PLA/toluene/water system revealed that MOPEO-PLA had a good surface activity almost equal to that of MOPEO. The MOPEO-PLA/PLA blend films were prepared by solvent casting on a water layer. Contact-angle measurements of MOPEO-PLA/PLA blend films confirmed that the hydrophilic PEO segments were selectivity accumulated at the oil/water interface. Moreover, the surface free energy on the 'water side' of the MOPEO-PLA/PLA blend films was increased because of the increase in polar components as a result of the ether bonds of the PEO segments. Schematic illustration of the adsorption property of a) MOPEO-PLA with a high molecular-weight PLA segment and b) MOPEO-PLA with a low-molecular-weight PLA segment at an ethyl acetate/water interface. PMID- 16245273 TI - UV-photocrosslinking of inulin derivatives to produce hydrogels for drug delivery application. AB - In this work, INU, a natural polysaccharide, has been chemically modified in order to obtain new photocrosslinkable derivatives. To reach this goal, INU has been derivatized with MA thus obtaining four samples (INU-MA derivatives) as a function of the temperature and time of reaction. An aqueous solution of the derivative INU-MA1 was irradiated by using a UV lamp with an emission range from 250 to 364 nm and without using photoinitiators. The obtained hydrogel showed a remarkable water affinity but it underwent a partial degradation in simulated gastric fluid. To overcome this drawback, INU-MA1 was derivatized with SA thus obtaining the INU-MA1-SA derivative designed to produce a hydrogel showing a low swelling and an increased chemical stability in acidic medium. Ibuprofen, as a model drug, was loaded by soaking into INU-MA1 and INU-MA1-SA hydrogels and its release from these matrices was evaluated in simulated gastrointestinal fluids. INU-MA1 hydrogel showed the ability to quickly release the entrapped drug thus indicating its potential as a matrix for an oral formulation. INU-MA1-SA hydrogel showed a pH-responsive drug delivery. Therefore it is a promising candidate for controlled drug release in the intestinal tract. PMID- 16245274 TI - Correlation between solid-state structures and enzymatic degradability of cocrystallized blends. AB - Solid-state structures and enzymatic degradability have been investigated for cocrystallized blends between poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [PHBV] and poly(3-hydroxybutyrate-co-3-hydroxypropionate) [PHBP]. From wide-angle X-ray diffraction patterns, small-angle X-ray scattering data, and the comparison of the enzymatic degradability of these blends, the solid-state structures of PHBV/PHBP blend samples, in which the PHBV component has higher isothermal crystal growth rate (G) value than the PHBP one, might be similar to those of the component PHBVs; while those of the PHBP/PHBV blend samples, in which PHBP component has higher G value, were similar to the component PHBPs. Normalized one dimensional correlation functions gamma(x) of PHBV/PHBP binary blends crystallized at 90 degrees C. PMID- 16245275 TI - Synthesis and rheological properties of polylactide/poly(ethylene glycol) multiblock copolymers. AB - Ring-opening polymerization of D,L-lactide was carried out in the presence of poly(ethylene glycol), using Zn powder as catalyst. The hydroxyl-capped PLA-PEG PLA triblock copolymers were coupled with adipoyl chloride at different molar ratios under mild conditions. N-Dimethylaminopyridine (DMAP) was used as catalyst of the coupling reaction. The resulting PLA/PEG multiblock copolymers were characterized by various analytical techniques such as IR, 1H NMR, SEC, and DSC. Sol-gel transition properties of the multiblock copolymers were investigated by mechanical rheology. The data showed that the sol-gel transition temperature and the transition modulus increased with increasing molecular weight and the solution concentration of the multiblock copolymers. [Graph: see text] Variation of storage modulus (G') and loss modulus (G'') as a function of temperature for a 20% sample of MB3. PMID- 16245276 TI - What to do? PMID- 16245277 TI - Bystin in perineural invasion of prostate cancer. AB - BACKGROUND: Bystin, Trophinin, and Tastin are component proteins of an adhesion molecule complex that plays a crucial role in the initial attachment of the embryo to the uterus. METHODS: Profiling of genes differentially expressed in the perineural invasion (PNI) in vitro model by gene microarray analysis showed overexpression of bystin in prostate cancer cells co-cultured with nerves. Validation was performed at the RNA levels using quantitative PCR. RESULTS: Bystin is overexpressed in cells co-cultured with nerves. Bystin is also present in human prostatic carcinoma (PCa) cells in PNI location in increasing gradient. Bystin is present in the supernatant of the PNI co-culture. CONCLUSIONS: Their adhesive and invasive functions in the trophoblast suggest that they might also play a role in perineural adhesion. Bystin is, therefore, an important therapeutic target for neurotropic cancers. PMID- 16245278 TI - 90K/Mac-2 binding protein is expressed in prostate cancer and induces promatrilysin expression. AB - BACKGROUND: 90K/Mac-2 binding protein is a cell adhesive protein whose level of expression has been correlated with metastatic potential in many different tumor types. The purpose of this investigation was to examine 90K expression in prostate cancer and to determine a possible role for 90K in cancer progression. METHODS: 90K expression in prostate cell lines and tissue samples was evaluated by immunohistochemistry. Expression in cell lines was also evaluated by Western blot analysis and real-time RT-PCR. Induction of promatrilysin by 90K was evaluated by ELISA. RESULTS: Some of the human prostate cell lines studied expressed 90K. 90K was over-expressed in 38.8% of prostate cancer tumor samples, 7.14% of PIN lesions, and 18.6% of normal tissue. 90K was also shown to induce promatrilysin expression in the prostate cell line, LNCaP. CONCLUSIONS: These data demonstrate that 90K is over-expressed in a large fraction of malignant tumors. The fact that 90K can induce expression of promatrilysin indicates a possible role for 90K in cancer progression and metastasis. This suggests that 90K over-expression may be a useful marker for examining prostate cancer progression. PMID- 16245279 TI - Prostate cancer and genetic susceptibility: a genome scan incorporating disease aggressiveness. AB - BACKGROUND: Prostate cancer is a heterogeneous disease, both genetically and phenotypically. Linkage studies attempting to map genes for hereditary prostate cancer (HPC) have proved challenging, and one potential problem contributing to this challenge is the variability in disease phenotypes. METHODS: We collected clinical data on 784 affected men with prostate cancer from 248 HPC families for whom a genomic screen was performed. Disease characteristics (i.e., Gleason score, stage, prostate-specific antigen (PSA)) were used to classify affected men into categories of clinically insignificant, moderate, or aggressive prostate cancer. To potentially enrich for a genetic etiology, we restricted linkage analyses to only men with aggressive disease, although we used genotype information from all family members; linkage analyses used both dominant and recessive models. In addition, subset analyses considered age at diagnosis, number of affected men per family and other stratifications to try to increase genetic homogeneity. RESULTS: Several regions of interest (heterogeneity LOD score, HLOD>1.0) were identified in families (n=123) with >or=2 affecteds with aggressive prostate cancer. "Suggestive" linkage was observed at chromosome 22q11.1 (Dominant model HLOD=2.18) and the result was stronger (Dominant HLOD=2.75) in families with evidence of male-to-male transmission. A second region at 22q12.3-q13.1 was also highlighted (Recessive model HLOD=1.90) among men with aggressive disease, as was a region on chromosome 18. CONCLUSIONS: These analyses suggest that using clinically defined phenotypes may be a useful approach for simplifying the locus heterogeneity problems that confound the search for prostate cancer susceptibility genes. PMID- 16245280 TI - Targeting of pericytes diminishes neovascularization and lymphangiogenesis in prostate cancer. AB - BACKGROUND: The walls of capillaries in prostate cancer are composed of endothelial cells, and pericytes. NG2 is a transmembrane proteoglycan on nascent pericytes with a functional role in neovascularization. METHODS: The anti angiogenic effect of hydron pellets containing NG2 neutralizing antibody was quantified in intracorneal PC-3 and LNCaP xenografts. TRAMP and TRAMP-C1 tumors grafted in NG2 knockout mice represented intrinsic pericyte targeting. TRAMP and TRAMP-C1 grafts were analyzed with confocal microscope for microvascular density (MVD) and lymphatic vascular density (LVD). RESULTS: NG2 neutralizing antibody decreased corneal neovascularization in PC3 (P<0.0001), and LNCaP (P=0.0079) xenografts. Mean MVD in TRAMP and TRAMP-C1 tumors in NG2 knockout mice were 71% (P=0.0006) and 63% (P=0.0011) lower than wild type controls, respectively. Mean LVD in TRAMP and TRAMP-C1 tumors in NG2 knockout mice were 73% (P=0.0003) and 84% (P<0.0001) lower than wild type controls, respectively. CONCLUSIONS: Targeting of pericyte-NG2 decreases neovascularization and lymphangiogenesis in prostate cancer significantly. PMID- 16245281 TI - Thrombin receptor expression is upregulated in prostate cancer. AB - BACKGROUND: Aberrant expression of protease-activated receptors (PARs) has been associated with increased angiogenesis, tumor growth, and metastasis of various cancers. We assessed the status of PAR1 expression in prostate cancer. METHODS: The study compared the abundance levels of PAR1 RNA and protein using real-time reverse-transcriptase polymerase chain reaction and immunoblotting in freshly resected prostate tissues from early localized-stage disease (n=9) to those from patients with advanced metastatic disease (n=7). PAR1 expression and localization was evaluated using immunohistochemical staining of prostate specimens with benign prostatic hyperplasia (n=27), early- (n=32) and advanced-stage (n=22) prostate cancer. Association analyses of PAR1 expression with expression of VEGF family of growth factors, their receptors, and clinicopathological characteristics of the patients were also performed. RESULTS: PAR1 RNA expression in advanced-stage prostate was 2.39-fold higher (P=0.024) and its protein expression was 2.75-fold higher (P=5.89x10(-5)) when compared with early-stage prostate cancer. PAR1 expression was localized to endothelial cells in vascular network of prostate tumor areas. The expression of PAR1 correlated statistically significantly with advanced disease stage (P=0.0006) and pre-operative PSA levels (P=0.005) in these samples. CONCLUSIONS: These findings demonstrate that PAR1 expression is increased in prostate cancer. Its predominant expression in vascular network suggests that it may play a direct and crucial role in angiogenesis and could be a relevant target for therapeutic interventions to control or to prevent disease progression. PMID- 16245282 TI - A selective retinoid X receptor agonist bexarotene (LGD1069, Targretin) prevents and overcomes multidrug resistance in advanced prostate cancer. AB - BACKGROUND: We previously reported that a retinoid X receptor agonist bexarotene prevented and overcame acquired drug resistance in advanced breast cancer and non small cell lung cancer. The present study was to evaluate the effect of bexarotene on the development of multidrug resistance in advanced prostate cancer. METHODS: Human prostate cancer cells PC3 were repeatedly treated in culture with paclitaxel, doxorubicin, or cisplatin with or without bexarotene for 3 months. Thereafter, cells were isolated and characterized for their drug sensitivity. RESULTS: Compared to parental cells, cells treated with a single therapeutic agent was resistant to the therapeutic agent, whereas cells treated with the combination remained chemosensitive. Cells with acquired drug resistance showed increased sensitivity to the cytotoxic agent when treated with the combination. Fluctuation analysis demonstrated that treatment with bexarotene decreased the rate of spontaneous development of drug resistance. These in vitro findings were further confirmed in the PC3 xenograft model. CONCLUSION: Our results suggest a role of bexarotene in combination with chemotherapeutic agents in prevention and overcoming acquired drug resistance in advanced prostate cancer. PMID- 16245283 TI - Antifungal and antibacterial properties of a silver-loaded cellulosic fiber. AB - The skin is the interface between the body and the environment. Each skin type has a specific skin physiology and is more or less adapted for protection against multiple stress factors. Textiles on the other hand are the tissues with the longest contact to the human skin. They play a critical role especially in skin conditions with an increased rate of bacterial and fungal infections like atopic dermatitis and hyperhidrosis, and in diabetic patients and aged skin. The present study demonstrates the antifungal and antibacterial effects of SeaCell Active in an in vitro test system against Candida albicans (DSM 11225), Candida tropicalis (ATCC 1169), and Candida krusei (ATCC 6258). Furthermore, the antibacterial activity of fibers with different amounts of SeaCell Active fibers in a dose dependent manner against Staphylococcus aureus (ATCC 22923) and Escherichia coli (ATCC 35218) could be demonstrated. If this fiber seems to be suited for bioactive textiles in specific anatomical regions and skin conditions with a susceptibility for fungal and bacterial infections due to Candida species, namely Staphylococcus aureus and Escherichia coli, must be examined by means of further investigations, especially in vivo tests in human, considering allergic and toxic effects of the fiber. PMID- 16245284 TI - Characterization of pegylated copolymeric micelles and in vivo pharmacokinetics and biodistribution studies. AB - The aim of this study was to evaluate the influence of pegylated copolymeric micelle carrier on the biodistribution of drug in rats. The copolymers were synthesized via a modified ring-opening copolymerization of lactone monomers (epsilon-caprolactone, delta-valerolactone, L-lactide) and poly(ethylene glycol) (PEG(10,000) and PEG(4000)). The molecular weights and the polydispersities of synthesized copolymers were in the range of 15,000-31,000 g/mol and 1.7-2.7, respectively. All of the pegylated amphiphilic copolymers were micelles formed with low CMC values in the range of 10(-7)-10(-8)M. The drug-loaded micelles were prepared via a dialysis method. The average particle size of micelles was around 150-200 nm. The cytotoxicity in terms of cell viability after treated with PCL PEG, PVL-PEG, and PLA-PEG micelles was insignificant. PCL-PEG and PVL-PEG micelles without branch side chain in structures had higher drug loading than PLA PEG micelles. In vitro release profiles indicated the release of indomethacin from these micelles exhibited a sustained release behavior. The similar phenomenon was also observed in vivo in rats. The pegylated copolymeric micelles not only decreased drug uptake by the liver and kidney, but also prolonged drug retention in the blood. PMID- 16245285 TI - Effects of adhesive systems and luting agents on bonding of fiber posts to root canal dentin. AB - The study evaluated the influence of different luting materials on the microtensile bond strength of glass fiber posts to root canal dentin. Thirty extracted maxillary premolars were endodontically treated, and the roots were prepared for post cementation using the FRC Postec system (Vivadent). Two luting materials (Multilink, Vivadent and Clearfil Photo Core, Kuraray) were used in combination with three adhesive: Multilink Primer (Vivadent), Clearfil Photo Bond, and Clearfil New Bond (Kuraray). A composite build-up was performed around the root to provide adequate gripping during testing. Specimens were cut to obtain beams with the post in the center and with the radicular dentin overlaid by the composite build-up on each side. Microtensile testing was performed with a universal testing machine at a cross-head speed of 0.5 mm/min. The failure mode was classified under a stereomicroscope and four representative beams of each group were selected for SEM analysis. Bond strength data that were analyzed with two-way ANOVA and Student-Newman-Keuls multiple comparisons tests revealed that adhesive systems, luting materials, and the interaction between these two factors significantly influenced the bond strength results (p < 0.01). Multilink applied with its own adhesive system obtained the best results, while the lowest bond strength was achieved with clearfil photo core in combination with multilink primer. PMID- 16245286 TI - Augmentation of implant purchase with bone cements: an in vitro study of injectability and dough distribution. AB - Vertebroplasty is widely used to treat (augment) osteoporotic fractures of the spine. This technique--with or without metallic implants--might have more widespread indications, if the mechanics of the injection and distribution of the cement dough through cannulated instruments and implants were better understood. This study was performed to investigate injectability of calcium phosphate and acrylic bone cements through implant prototypes, which featured different perforated sleeve designs. Using a custom-made capillary rheometer, the forces needed to inject 10 mL of the cement dough through standard cannulas were measured in the first series of experiments. In the second series, plastic sleeves were attached to the rheometer, simulating the implant. In both series, the dough was injected into ambient laboratory atmosphere, and in the second series, cement distribution was analyzed by means of an optical system. Injection of cement dough through the cannulas required forces between 50 and 400 N in the case of acrylic cements and between 40 and 500 N in case of the calcium phosphate cements. Using different sleeves did not have a significant influence on the distribution of the cement dough around the sleeve. The amount of cement dough injected was reduced when a perforated implant was attached to the cannula. More material was delivered through the proximal holes of the implant, leading to a V shaped distribution of the cement dough. Among topics to be investigated in future studies is determination of the injectability of cement dough into trabecular bone or bone-like structures. PMID- 16245287 TI - Chemical characteristics and cytocompatibility of collagen-based scaffold reinforced by chitin fibers for bone tissue engineering. AB - Chitin is a kind of seemly material to match PLLA for a scaffold, which may create an appropriate environment for the regeneration of tissues. In this study, we prepared and evaluated a new nano-hydroxyapatite/collagen/PLLA (nHACP) scaffold reinforced by chitin fibers for bone-tissue engineering. The chitin fibers were crosslinked with PLLA by dicyclohexylcarbodimide (DCC). The chemical characteristics were evaluated by Fourier transformed infrared (FTIR) spectroscopy and X-ray photoelectron spectroscopy (XPS). The mechanical strength was measured by compressive tests. The fibers, crosslinked with PLLA, could enhance the compressive strength of the scaffold about four times. Human marrow mesenchymal stem cells (MSCs) culture showed that the reinforced nHACP scaffolds were more cytocompatible than that without reinforcement. The crosslinks hardly affected the cytocompatibility of the reinforced scaffolds. The results suggested that the reinforced scaffolds (DCC crosslinked) might be a promising candidate for bone-tissue engineering. PMID- 16245288 TI - A clinical case report: interface analysis of a successful well-functioning transmandibular implant from a cadaver mandible. AB - Transmandibular implants (TMI) are indicated both for functional reconstruction of the severely atrophic mandible and when routine augmentation is unpredictable. This study investigates the interface of bone around a TMI, retrieved from the cadaver. The TMI had successfully functioned for 7 years. The mandible was immersed in 10% formaldehyde and sectioned into nine appropriate pieces. Samples were embedded in polymethylmethacrylate, and cut around the transosseous posts and cortical screws in both vertical and horizontal sections. Samples were analyzed at 400 MHz (nominal lateral resolution, 2.5 microm) using a UH3 Scanning Acoustic Microscope (Olympus, Tokyo, Japan). The middle of implant specimens 1-4 were cut to 50 microm, and stained by toluidine blue for light microscopy. Dental X-rays showed no bone resorption around any implant. On a 2-mm lateral scan, almost uniform interface space was seen between bone and implant surface in cortical screws. There are wider spaces around the transmandibular posts in the superior area. Histology revealed the small area of direct contact. There is bone marrow space in the interface, with no significant fibrous tissue. We interpret these results at the interface to be because of adaptation for stress distribution. PMID- 16245289 TI - In vitro studies on the influence of precultural conditioning method on osteoblast reactions of a new type of injectable calcium cement material. AB - A new injectable dicalcium phosphate dehydrate (DCPD)-based cement material "PD" VitalOs Cement was studied to elucidate the process of equilibrium occurring in the early stage of implantation. The present study investigated the pH variations of the cement sample-immersing culture medium at determined intervals, time dependent calcium/phosphate release, cell proliferation, and vitality in the cells-cement coculture milieu, after different preculture conditionings of the samples. Measurement of pH variation showed that without renewing the medium, pH value of sample lixiviate medium first dropped and, after 70 h, gradually balanced. When medium was renewed each day, pH value of lixiviate medium first descended and, after 24 h, gradually returned to pH 7.2. The cell viability revealed an excellent cytocompatibility of the cement. Both cell proliferation and vitality test showed that the preculture conditioning treatment is important at least for good performance of osteoblasts growing on the surface of calcium phosphate hydraulic cement (CPHC) samples in vitro. The results of calcium and phosphate assays clearly showed that this cement material can continuously dissolve to release calcium and phosphate in the liquid cell culture environment. The decrease of proliferation in some experimental groups with short conditioning is due to an excess of acid, which still can have some influence on cell growth after 24 h, since the biological milieu is not continuously renewed as in in vivo conditions. PMID- 16245290 TI - New oxidation treatment of NiTi shape memory alloys to obtain Ni-free surfaces and to improve biocompatibility. AB - Various oxidation treatments were applied to nearly equiatomic NiTi alloys so as to form a Ni-free protective oxide on the surface. Sample surfaces were analyzed by X-ray Photoelectron Spectroscopy, and NiTi transformation temperatures were determined by differential scanning calorimetry (DSC) before and after the surface treatment. An ion release experiment was carried out up to one month of immersion in SBF for both oxidized and untreated surfaces. The results show that oxidation treatment in a low-oxygen pressure atmosphere leads to a high surface Ti/Ni ratio, a very low Ni surface concentration and a thick oxide layer. This oxidation treatment does not significantly affect the shape memory properties of the alloy. Moreover, the oxide formed significantly decreases Ni release into exterior medium comparing with untreated surfaces. As a consequence, this new oxidation treatment could be of great interest for biomedical applications, as it could minimize sensitization and allergies and improve biocompatibility and corrosion resistance of NiTi shape memory alloys. PMID- 16245291 TI - Enhanced cell-seeding into 3D porous scaffolds by use of magnetite nanoparticles. AB - To engineer functional tissues, a large number of cells must be successfully seeded into scaffolds. We previously proposed a methodology for tissue engineering using magnetite nanoparticles and magnetic force, which we termed "Mag-TE." In the present study, we applied the Mag-TE technique to a cell seeding process and have termed the technique "Mag-seeding." The cell-seeding efficiency of NIH/3T3 fibroblasts (FBs) by Mag-seeding was investigated using six types of commercially available scaffolds (5 collagen sponges and 1 D,D-L,L polylactic acid sponge) having various pore sizes. FBs were magnetically labeled with our original magnetite cationic liposomes (MCLs), which have a positive surface charge, to improve adsorption onto the cell surface. FBs labeled with MCLs were seeded onto a scaffold, and a magnet (4 kG) was placed under the scaffold. Mag seeding facilitated successful cell seeding into the deep internal space of the scaffolds. Cell-seeding efficiency increased significantly in all scaffolds when compared to those without magnetic force. Moreover, when a high-intensity magnet (10 kG) was used, cell-seeding efficiency was significantly enhanced. These results suggest that Mag-seeding is a promising approach for tissue engineering. PMID- 16245292 TI - In vitro and in vivo evaluation of ultrananocrystalline diamond for coating of implantable retinal microchips. AB - In this work, ultrananocrystalline diamond (UNCD) thin films were evaluated for use as hermetic and bioinert coatings for a retinal microchip. These films were deposited on highly conductive Si substrates at different temperatures (from 400 to 800 degrees C), using microwave plasma enhanced chemical vapor deposition with argon-rich Ar/CH4 gas mixtures and different relative amounts of hydrogen (0 20%). Scanning electron microscopy studies showed that all the films are dense and continuous. Results of cyclic voltammetry test revealed that when there was <2% of hydrogen in the plasma, the film obtained renders the surface electrochemically inactive, with very low leakage currents ( approximately 4 x 10(-7) A/cm2 at +/-5 V). In addition, in vivo tests of the UNCD-coated Si samples were performed by implanting them in the eyes of rabbits for 4-6 months within the eye physiological environment. According to all these results, it was concluded that UNCD is a promising candidate for use as the encapsulating coatings for implantable retinal microelectronic devices. PMID- 16245293 TI - Our approach to pediatric dermatologic laser surgery. AB - BACKGROUND AND OBJECTIVES: Many pediatric dermatological conditions may be successfully treated with laser surgery. STUDY DESIGN/MATERIALS AND METHODS: The clinical approach to various pediatric dermatological conditions utilizing laser treatment options is discussed. RESULTS: Clinical uses of various modalities such as pulsed dye laser, KTP laser, Nd-YAG laser, Q-switched ruby laser, erbium-YAG laser, diode laser, non-coherent blue light sources, and fractional resurfacing are presented with successful parameters, developed over 22 years, utilized on a daily basis in a laser surgery clinic. CONCLUSION: Laser surgery can make a significant improvement in many pediatric skin lesions, thanks to the unique properties of pediatric skin and a vast array of laser technologies. PMID- 16245294 TI - Retraction: Effect of low-power He-Ne laser irradiation on rabbit articular chondrocytes in vitro. AB - Lasers in Surgery and Medicine regrets to inform readers that the article, "Effects of low-power He-NE laser irradiation on rabbit articular chondrocytes in vitro," by YL Jia and ZY Guo(Vol.34, No.4, pp. 323-328, 2004, DOI.10.1002/lsm.20017 is being retracted on account of concerns over the validity of the data reported therein based on further consideration by the journal's Editor. PMID- 16245295 TI - Intraoperative photodynamic therapy of the chest cavity in malignant pleural mesothelioma bearing rats. AB - BACKGROUND AND OBJECTIVE: Experimental assessment of anticancer effect, normal tissue damage, and toxicity of intrathoracic mTHPC-mediated photodynamic therapy (PDT) combined to surgery in malignant pleural mesothelioma (MPM) bearing rats. STUDY DESIGN/MATERIALS AND METHODS: Six days after implantation of syngenic malignant mesothelioma cells in the left chest cavity of Fischer rats (n = 21) and 4 days after sensitization (0.1 mg/kg mTHPC), a left-sided pneumonectomy was performed, followed by intraoperative light delivery (652 nm, fluence 20 J/cm(2)), either by spherical illumination of the chest cavity (fluence rate 15 mW/cm(2)) or by focal illumination of a tumor area (fluence rate 150 mW/cm(2)). Controls comprised tumor-bearing untreated animals, tumor-bearing animals undergoing pneumonectomy, and tumor-bearing animals undergoing pneumonectomy and light delivery without sensitization or sensitization without light delivery. No thoracocentesis was performed during follow-up. RESULTS: An invasively growing sarcomatous type of mesothelioma was found in all animals at day 10, without tumor necrosis in control animals. PDT resulted in 0.5-1 mm deep inhomogeneous tumor necrosis after spherical, and in a 1-2 mm deep tumor necrosis after focal illumination. No injury to mediastinal organs was observed, neither after PDT with spherical nor with focal light delivery except focal interstitial lung fibrosis at the mediastinal area of the opposite lung. All animals with pneumonectomy followed by spherical PDT of the entire tumor-bearing chest cavity died within 72 hours whereas all other animals survived. All animals that died presented massive pleural effusion. CONCLUSIONS: PDT following pneumonectomy in mesothelioma bearing rats was technically feasible and allowed to study its effect on tumor and normal tissues. PDT-related tumor necrosis was observed after spherical and focal light delivery, however, pneumonectomy followed by PDT with spherical light delivery to the tumor-bearing chest cavity resulted in fatal complications. PMID- 16245298 TI - Glycogen phosphorylase inhibitors: a free energy perturbation analysis of glucopyranose spirohydantoin analogues. AB - GP catalyzes the phosphorylation of glycogen to Glc-1-P. Because of its fundamental role in the metabolism of glycogen, GP has been the target for a systematic structure-assisted design of inhibitory compounds, which could be of value in the therapeutic treatment of type 2 diabetes mellitus. The most potent catalytic-site inhibitor of GP identified to date is spirohydantoin of glucopyranose (hydan). In this work, we employ MD free energy simulations to calculate the relative binding affinities for GP of hydan and two spirohydantoin analogues, methyl-hydan and n-hydan, in which a hydrogen atom is replaced by a methyl- or amino group, respectively. The results are compared with the experimental relative affinities of these ligands, estimated by kinetic measurements of the ligand inhibition constants. The calculated binding affinity for methyl-hydan (relative to hydan) is 3.75 +/- 1.4 kcal/mol, in excellent agreement with the experimental value (3.6 +/- 0.2 kcal/mol). For n-hydan, the calculated value is 1.0 +/- 1.1 kcal/mol, somewhat smaller than the experimental result (2.3 +/- 0.1 kcal/mol). A free energy decomposition analysis shows that hydan makes optimum interactions with protein residues and specific water molecules in the catalytic site. In the other two ligands, structural perturbations of the active site by the additional methyl- or amino group reduce the corresponding binding affinities. The computed binding free energies are sensitive to the preference of a specific water molecule for two well-defined positions in the catalytic site. The behavior of this water is analyzed in detail, and the free energy profile for the translocation of the water between the two positions is evaluated. The results provide insights into the role of water molecules in modulating ligand binding affinities. A comparison of the interactions between a set of ligands and their surrounding groups in X-ray structures is often used in the interpretation of binding free energy differences and in guiding the design of new ligands. For the systems in this work, such an approach fails to estimate the order of relative binding strengths, in contrast to the rigorous free energy treatment. PMID- 16245299 TI - Involvement of TRAIL/TRAIL-receptors in human intestinal cell differentiation. AB - Despite the fact that tumor necrosis factor (TNF)-related apoptosis inducing ligand (TRAIL) and its receptors (TRAIL-Rs) are expressed in intestinal mucosa, little is known about the biological role of this system in intestinal cell physiology. The expression of surface TRAIL and TRAIL-R1, -R2, -R3, -R4 were examined by flow cytometry in the immortalized human cell line tsFHI under culture conditions promoting growth or growth arrest and expression of differentiated traits. A progressive increase of surface TRAIL expression paralleled tsFHI differentiation, consistently with immunohistochemistry analysis showing an increase of TRAIL immunostaining along the crypt-villus axis in normal jejuneal mucosa. In spite of the presence of TRAIL-R1 and TRAIL-R2 "death receptors," recombinant TRAIL was not cytotoxic for tsFHI cells. Exposure of tsFHI to recombinant TRAIL rather increased/anticipated the expression levels of the cyclin-dependent kinase inhibitors p21 and p27, which mediate the induction of growth arrest and the stabilization of differentiated traits, respectively, as well as of the canonical differentiation marker DPPIV. The differentiation inducing activity of TRAIL was abolished by pre-incubation with a Fc-TRAIL-R2 chimera. On the other hand, TRAIL did not significantly modulate the levels of osteoprotegerin (OPG), CXCL8/IL-8, CXCL9/MIG, and CXCL10/IP10 spontaneously released or induced by inflammatory cytokines. Taken together, these data suggest that TRAIL might act as a paracrine trophic cytokine on intestinal epithelium, promoting intestinal cell differentiation. PMID- 16245300 TI - Involvement of protein phosphatase 2A in the maintenance of E-cadherin-mediated cell-cell adhesion through recruitment of IQGAP1. AB - Serine/threonine protein phosphatase (PP) 2A regulates many biological processes, however it remains unclear whether PP2A participates in cadherin-mediated cell cell adhesion. We show here that the core enzyme of PP2A (PP2A-AC) is localized in the cell-cell adhesion sites between adjacent cells and associated with the E cadherin-catenins complex in non-malignant human mammary epithelial (HME) cells at confluence. Treatment of the cells with either okadaic acid (OA), an inhibitor of PP2A, or siRNA for the regulatory subunit A of PP2A (PP2A-A) caused disruption of cell-cell adhesion and F-actin assembly, without affecting the complex formation of E-cadherin with beta- and alpha-catenins. While a small GTPase Rac and its effector IQGAP1 were associated with the E-cadherin-catenins complex, either OA or PP2A-A siRNA concomitantly induced the dissociation of IQGAP1, but not Rac, from the complex and the internalization of E-cadherin from the cell surface. We therefore propose that PP2A plays a crucial role in the maintenance of cell-cell adhesion through recruitment of IQGAP1 to the Rac-bound E-cadherin catenins complex. PMID- 16245301 TI - Implication of the hypoxia response element of the Vegf promoter in mouse models of retinal and choroidal neovascularization, but not retinal vascular development. AB - Retinal neovascularization (NV) and macular edema, resulting from blood-retinal barrier (BRB) breakdown, are major causes of visual loss in ischemic retinopathies. Choroidal NV (CNV) occurs in diseases of the retinal pigmented epithelium/Bruch's membrane complex and is another extremely prevalent cause of visual loss. We used mice in which the hypoxia response element (HRE) is deleted from the vascular endothelial growth factor (vegf) promoter (Vegf(delta/delta) mice) to explore the role of induction of VEGF through the HRE in these disease processes. Compared to wild type (Vegf+/+) mice with oxygen-induced ischemic retinopathy (OIR) in which vegf mRNA levels were increased and prominent retinal NV and BRB breakdown occurred, Vegf(delta/delta) littermates with OIR failed to increase vegf mRNA levels in the retina and had significantly less retinal NV and BRB breakdown, but showed prominent dilation of some superficial retinal vessels. Vegf(+/delta) littermates with ischemic retinopathy developed comparable retinal NV to Vegf+/+ mice, exhibited intermediate levels of BRB breakdown, and did not show vasodilation. In a mouse model of CNV, due to laser-induced rupture of Bruch's membrane, the area of CNV at Bruch's membrane rupture sites was more than tenfold greater in Vegf+/+ mice than in Vegf(delta/delta) littermates. In contrast to these dramatic differences in pathologic ocular NV, Vegf(delta/delta) mice showed subtle differences in retinal vascular development compared to Vegf+/+ mice; it was slightly delayed, but otherwise normal. These data suggest that induction of VEGF through the HRE in its promoter is critical for retinal and CNV, but not for retinal vascular development. PMID- 16245302 TI - Transcriptomic analysis of an in vitro murine model of ovarian carcinoma: functional similarity to the human disease and identification of prospective tumoral markers and targets. AB - Ovarian cancer is an aggressive disease of poor prognostic when detected at advanced stage. It is widely accepted that the ovarian surface epithelium plays a central role in disease etiology, but little is known about disease progression at the molecular level. To identify genes involved in ovarian tumorigenesis, we carried out a genome-wide transcriptomic analysis of six spontaneously transformed mouse ovarian surface epithelial (MOSE) cell lines, an in vitro model for human ovarian carcinoma. Loess normalization followed by statistical analysis with control of multiple testing resulted in 509 differentially expressed genes using an adjusted P-value < or = 0.05 as cut-off. The top 20 differentially expressed genes included 10 genes (Spp1, Cyp1b1, Btg1, Cfh, Mt1, Mt2, Igfbp5, Gstm1, Gstm2, and Esr1) implicated in various aspects of ovarian carcinomas, and other 3 genes (Gsto1, Lcn7, and Alcam) associated to breast cancer. Upon functional analysis, the majority of alterations affected genes involved in glutathione metabolism and MAPK signaling pathways. Interestingly, over 20% of the aberrantly expressed genes were related to extracellular components, suggestive of potential markers of disease progression. In addition, we identified the genes Pura, Cnn3, Arpc1b, Map4k4, Tgfb1i4, and Crsp2 correlated to in vivo tumorigenic parameters previously reported for these cells. Taken together, our findings support the utility of MOSE cells in studying ovarian cancer biology and as a source of novel diagnostic and therapeutic targets. PMID- 16245303 TI - CRH functions as a growth factor/cytokine in the skin. AB - We tested the effect of CRH and related peptides in a large panel of human skin cells for growth factor/cytokine activities. In skin cells CRH action is mediated by CRH-R1, a subject to posttranslational modification with expression of alternatively spliced isoforms. Activation of CRH-R1 induced generation of both cAMP and IP3 in the majority of epidermal and dermal cells (except for normal keratinocytes and one melanoma line), indicating cell type-dependent coupling to signal transduction pathways. Phenotypic effects on cell proliferation were however dependent on both cell type and nutrition conditions. Specifically, CRH stimulated dermal fibroblasts proliferation, by increasing transition from G1/0 to the S phase, while in keratinocytes CRH inhibited cell proliferation. In normal and immortalized melanocytes CRH effect showed dichotomy and thus, it inhibited melanocyte proliferation in serum-containing medium CRH through G2 arrest, while serum free media led instead to CRH enhanced DNA synthesis (through increased transition from G1/G0 to S phase and decreased subG1 signal, indicating DNA degradation). CRH also induced inhibition of early and late apoptosis in the same cells, demonstrated by analysis with the annexin V stains. Thus, CRH acts on epidermal melanocytes as a survival factor under the stress of starvation (anti apoptotic) as well as inhibitor of growth factors induced cell proliferation. In conclusion, CRH and related peptides can couple CRH-R1 to any of diverse signal transduction pathways; they also regulate cell viability and proliferation in cell type and growth condition-dependent manners. PMID- 16245304 TI - Panic disorder and suicidal ideation in primary care. AB - The purpose of this study was to ascertain whether panic disorder (PD) and suicidal ideation are associated in an inner-city primary care clinic and whether this association remains significant after controlling for commonly co-occurring psychiatric disorders. We surveyed 2,043 patients attending a primary care clinic using the Primary Care Evaluation of Mental Disorders (PRIME-MD) Patient Health Questionnaire, a screening instrument that yields provisional diagnoses of selected psychiatric disorders. We estimated the prevalence of current suicidal ideation and of common psychiatric disorders including panic disorder and major depression. A provisional diagnosis of current PD was received by 127 patients (6.2%). After adjusting for potential confounders (age, gender, major depressive disorder [MDD], generalized anxiety disorder, and substance use disorders), patients with PD were about twice as likely to present with current suicidal ideation, as compared to those without PD (adjusted odds ratio [AOR] = 1.84; 95% confidence interval [CI]: 1.06-3.18; P = .03). After adjusting for PD and the above-mentioned potential confounders, patients with MDD had a sevenfold increase in the odds of suicidal ideation, as compared to those without MDD (AOR = 7.00; 95% CI: 4.42-11.08; P < .0001). Primary care patients with PD are at high risk for suicidal ideation, and patients with PD and co-occurring MDD are at especially high risk. PD patients in primary care thus should be assessed routinely for suicidal ideation and depression. PMID- 16245305 TI - Effect of laminin-1 on intestinal cell differentiation involves inhibition of nuclear nucleolin. AB - Intestinal epithelial cells are characterized by continuous renewal and differentiation events, which may be influenced by the basement membrane, and in particular laminins, which are major components of this specialized extracellular matrix. The function and signaling pathways of laminins in these processes are still poorly documented. In this study, we investigated the possible role and the subcellular localization of nucleolin, a nuclear shuttling protein, in relation to differentiation of human intestinal epithelial Caco2/TC7 cells triggered by exogenous laminin-1. Immunofluorescence and Western blot analysis indicated that laminin-1 induced early differentiation of the cells concomitantly to a decrease in nuclear nucleolin and its a cell surface location. We also showed that both effects of laminin-1 on Caco2/TC7 cells--induction of the differentiation marker sucrase-isomaltase and redistribution of nucleolin--could be mediated by a beta1 integrin dependent cascade that implicated activation of the p38 MAPK pathway. In addition, knock-down of nucleolin expression by the small interfering RNA strategy mimicked the effect of laminin-1 as it resulted in the induction of cell polarization and differentiation. Thus, our study suggests that changes in the subcellular distribution and expression level of nucleolin play an important role in intestinal cell differentiation and relay the signaling pathway induced by laminin-1. PMID- 16245306 TI - Stable knockdown of CFTR establishes a role for the channel in P2Y receptor stimulated anion secretion. AB - P2Y receptor regulation of anion secretion was investigated in porcine endometrial gland (PEG) epithelial cells. P2Y2, P2Y4, and P2Y6 receptors were detected in monolayers of PEG cells and immunocytochemistry indicated that P2Y4 receptors were located in the apical membrane. Apical membrane current measurements showed that Ca2+-dependent and PKC-dependent Cl- channels were activated following treatment with uridine triphosphate (UTP) (5 microM). Current voltage relationships comparing calcium-dependent and PKC-dependent UTP responses under biionic conditions showed significant differences in selectivity between Cl )and I- for the PKC-dependent conductance (P(I)/P(Cl) = 0.76), but not for Ca2+ dependent conductance (PI/P(Cl) = 1.02). The I-/Cl- permeability ratio for the PKC-dependent conductance was identical to that measured for 8-cpt cAMP. Furthermore, PKC stimulation using phorbol 12-myristate 13-acetate (PMA) activated an apical membrane Cl- conductance that was blocked by the CFTR selective inhibitor, CFTRinh-172. CFTR silencing, accomplished by stable expression of small hairpin RNAs (shRNA), blocked the PKC-activated conductance associated with UTP stimulation and provided definitive evidence of a role for CFTR in anion secretion. CFTR activation increased the initial magnitude of Cl- secretion, and provided a more sustained secretory response compared to conditions where only Ca2+-activated Cl- channels were activated by UTP. Measurements of [cAMP]i following UTP and PMA stimulation were not significantly different than untreated controls. Thus, these results demonstrate that UTP and PMA activation of CFTR occurs independently of increases in intracellular cAMP and extend the findings of earlier studies of CFTR regulation by PKC in Xenopus oocytes to a mammalian anion secreting epithelium. PMID- 16245308 TI - Characterisation of serum-induced intracellular Ca2+ oscillations in primary bone marrow stromal cells. AB - Intracellular Ca2+ signalling is pivotal to cell function and [Ca2+]i oscillations permit precise and prolonged modulation of an array of Ca2+ sensitive processes without the need for extended, global elevations in [Ca2+]i. We have studied [Ca2+]i signalling in primary rat marrow stromal cells exposed to foetal calf serum (FCS) constituents at concentrations up to those required to promote growth and differentiation in culture. Spontaneous [Ca2+]i signalling was not observed, but exposure to 1% FCS induced regular, sustained Ca2+ oscillations in 41 +/- 3% of cells. Incidence of FCS-induced oscillations was dose-dependent, saturating at 0.5%. These oscillations were arrested by disruption of Ca2+ stores with 100 nM-1 microM thapsigargin or discharge of mitochondrial membrane potential and were sensitive to blockade of IP3-receptors by 50 microM 2-amino ethoxydiphenyl borate (2-APB) and inhibition of phospholipase C with 5 microM U73122. The oscillations decreased in frequency and amplitude following inhibition of Ca2+ influx with EGTA or La3+ but were poorly sensitive to nifedipine (1-10 microM) and Bay K 8644 (300 nM). The factor(s) responsible for inducing [Ca2+]i oscillations are heat stable, insensitive to disulphide bond reduction with 20 mM dithioerythritol and retained by a 30 kDa molecular weight filter. Serum is routinely present in culture medium at 10%-15% [v/v] and marrow stromal cells maintained under culture conditions exhibited sustained oscillations. This is the first demonstration of agonist-induced complex Ca2+ signals in marrow stromal cells. We conclude that Ca2+ oscillations occur constantly in these cells in culture and are potentially important regulators of cell proliferation and differentiation. PMID- 16245307 TI - TNF-alpha-related apoptosis-inducing ligand decoy receptor DcR2 is targeted by androgen action in the rat ventral prostate. AB - The apoptotic cell death process in the prostate is known to be under the control of androgens. Tumor necrosis factor-alpha (TNF-alpha)-related apoptosis-inducing ligand (TRAIL) is a member of the TNF-alpha family of cytokines, known to induce apoptosis upon binding to its death domain-containing receptors, DR4/TRAIL-R1 and DR5/TRAIL-R2. Two additional TRAIL receptors, DcR1/TRAIL-R3 and DcR2/TRAIL-R4, lack functional death domains and act as decoy receptors for TRAIL. In this study, we examined whether TRAIL and cellular receptors expression was targeted by androgens during the apoptotic cell death process in the hormone sensitive ventral prostate. The role of androgens was investigated using two sets of experiment. (1) Androgen deprivation associated with an apoptotic process resulted in a decrease in DcR2 mRNA and protein expression in the ventral prostate 3 days after castration. Testosterone administration to castrated adult rats prevented the decrease in DcR2 mRNA and protein levels in the ventral prostate. In contrast, DcR2 expression was modified, neither in the dorsolateral nor in the anterior prostate following castration. No changes were observed in DR4, DR5, DcR1, and TRAIL mRNA and protein levels in prostate after castration. (2) A specific decrease in DcR2 expression was observed in the ventral prostate after treatment of rats with the anti-androgen flutamide. Together, the present results suggest that testosterone specifically controls DcR2 expression in the adult rat ventral prostate. Androgen withdrawal, by reducing DcR2 expression, might leave the cells vulnerable to cell death signals generated by TRAIL via its functional receptors. PMID- 16245309 TI - MyoD recruits the cdk9/cyclin T2 complex on myogenic-genes regulatory regions. AB - During skeletal myogenesis, muscle-regulatory factors bHLH and MEF2 promote the expression of muscle-specific genes by recruiting several chromatin-modifying complexes on specific DNA regulatory sequences. A number of MyoD-interacting proteins have been reported, but whether they are recruited to the chromatin of myogenic loci, and the relationship with other chromatin bound proteins is unknown. We show that MyoD recruits cdk9/cyclin T2, together with the histone acetyltransferases p300 and PCAF, and the chromatin remodeling complex SWI/SNF, on promoters and enhancers of muscle-specific genes, and that this event correlates with the acetylation of histone tails, remodeling of chromatin, and phosphorylation of the C-terminal domain (CTD) of the RNA polymerase II at these elements. PMID- 16245310 TI - Characterization of pathologic complete response after preoperative chemoradiotherapy in carcinoma of the esophagus and outcome after pathologic complete response. AB - BACKGROUND: The purpose of the current study was to test the hypothesis that a lower clinical TNM stage is associated with a higher rate of pathologic complete response (pathCR) in patients with esophageal carcinoma receiving preoperative chemoradiotherapy and to determine whether outcome after pathCR is related to clinical stage or treatment. METHODS: Clinical parameters and surgical specimens of patients with esophageal carcinoma undergoing preoperative chemoradiotherapy were analyzed to identify predictors of pathCR. In patients with pathCR, predictors of overall survival (OS), disease-free survival (DFS), and distant recurrence were studied. RESULTS: Sixty-nine (29%) of 235 patients achieved pathCR. In patients with American Joint Committee on Cancer (AJCC) Stage II carcinoma, the proportion achieving pathCR was significantly larger than that achieving tolmetin > fenofibrate. The examined compounds inhibited the HO(.)-dependent deoxyribose degradation and scavenged the ROO(.) concentration dependently with an order of potencies similar to that of the superoxide radical system. Hence, these results indicate that the studied drugs show broad ROS scavenging property and, as a consequence, might decrease tissue damage due to the ROS and thus to contribute to anti-inflammatory therapy. PMID- 16245332 TI - DPATJ plays a role in retinal morphogenesis and protects against light-dependent degeneration of photoreceptor cells in the Drosophila eye. AB - The establishment of apicobasal polarity in epithelial cells is a prerequisite for their function. Drosophila photoreceptor cells derive from epithelial cells, and their apical membranes undergo elaborate differentiation during pupal development, forming photosensitive rhabdomeres and associated stalk membranes. Crumbs (Crb), a transmembrane protein involved in the maintenance of epithelial polarity in the embryo, defines the stalk as a subdomain of the apical membrane. Crb organizes a complex composed of several PDZ domain-containing proteins, including DPATJ (formerly known as Discs lost). Taking advantage of a DPATJ mutant line in which only a truncated form of the protein is synthesized, we demonstrate that DPATJ is necessary for the stability of the Crb complex at the stalk membrane and is crucial for stalk membrane development and rhabdomere maintenance during late pupal stages. Moreover, DPATJ protects against light induced photoreceptor degeneration. PMID- 16245329 TI - Transforming growth factor-beta stimulates epithelial-mesenchymal transformation in the proepicardium. AB - The proepicardium (PE) migrates over the heart and forms the epicardium. A subset of these PE-derived cells undergoes epithelial-mesenchymal transformation (EMT) and gives rise to cardiac fibroblasts and components of the coronary vasculature. We report that transforming growth factor-beta (TGFbeta) 1 and TGFbeta2 increase EMT in PE explants as measured by invasion into a collagen gel, loss of cytokeratin expression, and redistribution of ZO1. The type I TGFbeta receptors ALK2 and ALK5 are both expressed in the PE. However, only constitutively active (ca) ALK2 stimulates PE-derived epithelial cell activation, the first step in transformation, whereas caALK5 stimulates neither activation nor transformation in PE explants. Overexpression of Smad6, an inhibitor of ALK2 signaling, inhibits epithelial cell activation, whereas BMP7, a known ligand for ALK2, has no effect. These data demonstrate that TGFbeta stimulates transformation in the PE and suggest that ALK2 partially mediates this effect. PMID- 16245333 TI - Characteristics of initiation and early events for muscle development in the Xenopus limb bud. AB - In Xenopus laevis, limb buds start to develop at a later point of the larval stage, prior to metamorphosis. This onset of limb development in Xenopus is totally different from that in amniotes such as birds and mammals, in which limb buds emerge at an early stage of embryogenesis, in parallel with other organogenesis. We investigated limb myogenesis in Xenopus, focusing on myogenic gene expression, myogenic ability of limb bud cells in the early stage, and the origin of myogenic precursor cells in the limb bud. The Xenopus early limb bud contains myoD/cardiac actin-positive and pax3/pax7-negative cells. Interestingly, results of transplantation experiments have revealed that this early limb bud contains myogenic precursor cells. In order to know the contribution of myogenic cells in somites to myogenic precursor cells in the early limb bud, we used a Cre LoxP system for tracing over a long period. The results of fate tracing for myogenic cells in somites of the Xenopus embryo suggested that early-specified myogenic cells in somites do not contribute to limb muscle in Xenopus. Taken together, the results suggest that limb muscle development in Xenopus has characteristics of initiation and early events distinct from those of other vertebrate clades. PMID- 16245334 TI - Distinct regulatory elements mediate the dynamic expression pattern of Nkx3.1. AB - Loss of Nkx3.1 function in mice results in defects in prostate development and epithelial hyperplasia, indicating that this gene plays important roles in both the initiation and maintenance of prostate differentiation. In humans, decreased NKX3.1 expression is associated with the progression of prostate cancer. Despite these roles in prostate development and disease, the transcriptional regulation of Nkx3.1 has not been systematically addressed. A reporter gene approach in transgenic mice was used to identify regulatory regions that dictate the expression pattern of Nkx3.1. A 32-kb DNA fragment from the Nkx3.1 locus that specifies the expected expression pattern during embryogenesis and postnatal life has been identified. Deletion analyses demonstrated that cis-regulatory elements that mediate expression in distinct sites are separable. A 5-kb fragment downstream of the Nkx3.1 coding region contains elements that support expression in the prostate and bulbourethral glands, whereas an upstream fragment contains elements that direct expression in somites and testes. Reporter gene expression analyses also revealed several previously unknown sites of Nkx3.1 expression in males, including urethral glands, glandular cells in the urethral diverticulum and basal epithelial cells in the prostate. In addition, these analyses revealed Nkx3.1 expression in female urethral glands. The identification of Nkx3.1 cis regulatory elements provides a unique starting point to dissect signaling pathways involved in prostate organogenesis and pathogenesis and provides a system to perturb gene expression throughout prostate development. PMID- 16245335 TI - Differentiation of cardiac Purkinje fibers requires precise spatiotemporal regulation of Nkx2-5 expression. AB - Nkx2-5 gene mutations cause cardiac abnormalities, including deficits of function in the atrioventricular conduction system (AVCS). In the chick, Nkx2-5 is elevated in Purkinje fiber AVCS cells relative to working cardiomyocytes. Here, we show that Nkx2-5 expression rises to a peak as Purkinje fibers progressively differentiate. To disrupt this pattern, we overexpressed Nkx2-5 from embryonic day 10, as Purkinje fibers are recruited within developing chick hearts. Overexpression of Nkx2-5 caused inhibition of slow tonic myosin heavy chain protein (sMHC), a late Purkinje fiber marker but did not affect Cx40 levels. Working cardiomyocytes overexpressing Nkx2-5 in these hearts ectopically up regulated Cx40 but not sMHC. Isolated embryonic cardiomyocytes overexpressing Nkx2-5 also displayed increased Cx40 and suppressed sMHC. By contrast, overexpression of a human NKX2-5 mutant did not effect these markers in vivo or in vitro, suggesting one possible mechanism for clinical phenotypes. We conclude that a prerequisite for normal Purkinje fiber maturation is precise regulation of Nkx2-5 levels. PMID- 16245336 TI - Developmental expression analysis of the mouse and chick orthologues of IRF6: the gene mutated in Van der Woude syndrome. AB - Development of the lip and palate involves a complex series of events that are frequently disturbed resulting in the congenital anomalies cleft lip and cleft palate. Van der Woude syndrome (VWS) is an autosomal dominant disorder that is characterised by cleft lip, cleft palate, lower lip pits, and hypodontia. VWS arises as the result of mutations in the gene encoding interferon regulatory factor 6 (IRF6). To provide insights into the role of IRF6 during embryogenesis, we have analysed the expression of this molecule during mouse and chick facial development. Irf6 was expressed in the ectoderm covering the facial processes during their fusion to form the upper lip and primary palate in both mouse and chick. However, while Irf6 was expressed in the medial edge epithelia of the developing secondary palate of the mouse, which fuses as in man, Irf6 was not expressed in the medial edge epithelia of the naturally cleft chick secondary palate. Similarly, Irf6 was found to be down-regulated in the medial edge epithelia of transforming growth factor beta3-null mice, which also exhibit cleft palate. Together, these results support a role for IRF6 during the fusion events that occur during development of the lip and palate. PMID- 16245338 TI - Notch1 signals through Jagged2 to regulate apoptosis in the apical ectodermal ridge of the developing limb bud. AB - The Notch family of receptors is involved in a wide variety of developmental processes, including cell fate specification, cell proliferation, and cell survival decisions during cell differentiation and tissue morphogenesis. Notch1 and Notch ligands are expressed in the developing limbs, and Notch signalling has been implicated in the formation of a variety of tissues that comprise the limb, such as the skeleton, musculature, and vasculature. Notch signalling has also been implicated in regulating overall limb size. We have used a conditional allele of Notch1 in combination with two different Cre transgenic lines to delete Notch1 function either in the limb mesenchyme or in the apical ectodermal ridge (AER) and limb ectoderm. We demonstrate that Notch signalling, involving Notch1 and Jagged2, is required to regulate the number of Fgf8-expressing cells that comprise the AER and that regulation of the levels of fibroblast growth factor signalling is important for the freeing of the digits during normal limb formation. Regulation of the extent of the AER is achieved by Notch signalling positively regulating apoptosis in the AER. We also demonstrate that Notch1 is not required for proper formation of all the derivatives of the limb mesenchyme. PMID- 16245339 TI - Analysis of the regulation of lin-41 during chick and mouse limb development. AB - We have cloned the chicken and mouse orthologues of the Caenorhabditis elegans heterochronic gene lin-41. During limb development, lin-41 is expressed in three phases over developmental time and most notably is associated with the developing autopod. Using chicken and mouse mutants and bead implantations, we report that lin-41 is genetically and biochemically downstream of both the Shh and Fgf signaling pathways. In C. elegans, it is proposed that lin-41 activity is temporally regulated by miRNAs (let-7 and lin-4) that bind to complementary sites in the lin-41 3'-untranslated region (UTR). Taking a bioinformatics approach, we also report the presence of potential miRNA binding sites in the 3'-UTR of chicken lin-41, including sites for the chicken orthologues of both C. elegans let-7 and lin-4. Finally, we show that these miRNAs and others are expressed in the chick limb consistent with the hypothesis that they regulate chicken Lin-41 activity in vivo. PMID- 16245337 TI - An in vitro model for characterizing the post-migratory cranial neural crest cells of the first branchial arch. AB - The cranial neural crest (CNC) is a transient cell population that originates at the crest of the neural fold and gives rise to multiple cell types during craniofacial development. Traditionally, researchers have used tissue explants, such as the neural tube, to obtain primary neural crest cells for their studies. However, this approach has inevitably resulted in simultaneous isolation of neural and non-neural crest cells as both of these cells migrate away from tissue explants. Using the Wnt1-Cre/R26R mouse model, we have obtained a pure population of neural crest cells and established a primary CNC cell culture system in which the cell culture medium best supports the proliferation of E10.5 first branchial arch CNC cells and maintains these cells in their undifferentiated state. Differentiation of CNC cells can be initiated by switching to a differentiation medium. In this model, cultured CNC cells can give rise to neurons, glial cells, osteoblasts, and other cell types, faithfully mimicking the differentiation process of the post-migratory CNC cells in vivo. Taken together, our study shows that the Wnt1-Cre/R26R mouse first branchial arch provides an excellent model for obtaining post-migratory neural crest cells free of any mesodermal contaminants. The cultured neural crest cells are under sustained proliferative, undifferentiated, or lineage-enhanced conditions, hence, serving as a tool for the investigation of the regulatory mechanism of CNC cell fate determination in normal and abnormal craniofacial development. PMID- 16245340 TI - The Protein Circular Dichroism Data Bank (PCDDB): a bioinformatics and spectroscopic resource. AB - This article describes the development and creation of the Protein Circular Dichroism Data Bank (PCDDB), a deposition and searchable data bank for validated circular dichroism spectra located at http://pcddb.cryst.bbk.ac.uk/. PMID- 16245341 TI - Extrapulmonary tuberculosis active infection misdiagnosed as cancer: Mycobacterium tuberculosis disease in patients at a Comprehensive Cancer Center (2001-2005). AB - BACKGROUND: Extrapulmonary tuberculosis is an uncommon disease in the U.S., even in immunosuppressed cancer patients. This study evaluated characteristics and frequency of extrapulmonary tuberculosis in patients at a tertiary care referral cancer center. METHODS: The records of all consecutive patients with Mycobacterium tuberculosis diagnosed during January 2001 through April 2005 at the M. D. Anderson Cancer Center were reviewed after obtaining institutional review board approval. RESULTS: There were 26 patients with active tuberculosis during the period studied; 18 of them were cancer patients and the others had been referred for a presumed cancer but did not have cancer. The overall rate of active tuberculosis during this period was 0.2 in 1000 new cancer diagnoses. There were 18 men (69%), the median age was 54 years (range, 3-84 yrs), and 16 patients (62%) were born in the U.S. Thirteen (72%) of the 18 cancer patients had solid-organ tumors; 3 of the 5 patients with a hematologic malignancy had non Hodgkin lymphoma. Three patients (12%) had diabetes mellitus, and 2 patients (8%) had received high-dose (>1 mg/kg of prednisone daily) corticosteroids in the previous week. No patient had a recent history (within the past 4 wks) of chemotherapy; 4 patients had neutropenia. Cough was a prominent symptom (31%), followed by bone pain (19%), dyspnea (15%), and fever (12%). Fifteen patients (58%) had extrapulmonary infection, including 5 patients with concurrent pulmonary involvement; 7 noncancer patients (88%) and 8 cancer patients (44%, P = 0.22) had extrapulmonary disease. In 11 patients (42%), the lungs were the only site of active tuberculosis. Cavitary pneumonia was seen radiographically in 3 of 16 patients (19%) with pulmonary tuberculosis. All M. tuberculosis isolates were susceptible to isoniazid, rifampin, ethambutol, and pyrazinamide; streptomycin resistance was noted in 1 of 22 (5%) isolates tested. Twenty-two patients (85%) received appropriate antituberculosis treatment; all had a clinical and radiographic response. In 3 patients (12%) the cause of death was attributed to M. tuberculosis disease; 2 of 18 cancer patients (11%) died of progressive M. tuberculosis, and they had advanced solid-organ cancer, whereas 1 of 8 patients (13%) without cancer died and the tuberculosis diagnosis was made only on postmortem examination. Univariate analysis showed no significant differences in patients or disease characteristics between non-U.S.-born and U.S.-born patients, whereas noncancer patients (age 52 yrs) and those with extrapulmonary tuberculosis (age 53 yrs) were younger compared with cancer patients (63 yrs; P < 0.007) and those with pulmonary disease (age 60 yrs; P = 0.09). CONCLUSIONS: Extrapulmonary tuberculosis was relatively common in younger patients with active M. tuberculosis infection, and was often initially misdiagnosed as cancer. PMID- 16245342 TI - Mutation in the delta-subunit of the nAChR suppresses the muscle defects caused by lack of Dystrophin. AB - Normal motility of the zebrafish embryo requires a large number of gene loci, many of which have human orthologues implicated in myasthenias and other myopathies. We have identified a mutation in the zebrafish that abolishes body motility. Embryos have narrower myofibrils and lack clusters of nicotinic acetylcholine receptors (nAChRs) on the surface of the somitic muscle. We mapped the mutation to the delta-subunit of the nAChR, showing this mutant to be a new allele of the previously named sofa potato (sop). The mutant allele carries a missense mutation in the extracellular domain altering the cysteine at position 150 to an arginine. The delta-subunit is expressed in all striated muscles in embryonic and early larval stages together with the alpha1, beta1, epsilon, and gamma-subunits of nAChR. In contrast to mammals that show switching from the gamma embryonic to the adult epsilon-subunit, the two subunits are coexpressed in zebrafish embryos. We, furthermore, demonstrated that the sop/delta-nAChR mutation is a suppressor of the myopathy caused by lack of Dystrophin. The myofiber detachment phenotype of Dystroglycan-deficient embryos was not suppressed, suggesting that Dystrophin and Dystroglycan play distinct roles in muscle formation and maintenance of muscle integrity. PMID- 16245343 TI - Olfactory sensitivity for aliphatic alcohols and aldehydes in spider monkeys (Ateles geoffroyi). AB - Using a conditioning paradigm, the olfactory sensitivity of five spider monkeys for homologous series of aliphatic 1-alcohols (1-propanol to 1-octanol) and n aldehydes (n-butanal to n-nonanal) was investigated. With the exception of 1 propanol, the animals significantly discriminated concentrations below 1 ppm from the odorless solvent, and in several cases, individual monkeys even demonstrated detection thresholds below 10 ppb. The results showed 1) spider monkeys to have a well-developed olfactory sensitivity for both substance classes, which for the majority of alcohols tested matches or even is better than that of the rat, and 2) a significant negative correlation between perceptibility in terms of olfactory detection thresholds and carbon chain length of the alcohols, but not of the aldehydes tested. These findings lend further support to the growing body of evidence suggesting that between-species comparisons of the number of functional olfactory receptor genes or of neuroanatomical features are poor predictors of olfactory performance, and that general labels such as "microsmat" or "macrosmat" (which are usually based on allometric comparisons of olfactory brain structures) are inadequate to describe a species' olfactory capabilities. PMID- 16245344 TI - Investigation into the usability of geometric morphometric analysis in assessment of sexual dimorphism. AB - Understanding sexual dimorphism is very important in studies of human evolution and skeletal biology. Sexual dimorphic characteristics can be studied morphologically and metrically, although morphologic studies pose several problems such as difficulties with quantification and interobserver error. Geometric morphometrics is a relatively new method that allows better assessment of morphologic characteristics. This paper aims to investigate the usability of this method by assessing three different morphologic characteristics in a sample of South African blacks: shape of the greater sciatic notch, mandibular ramus flexure, and shape of the orbits. Relative warps, thin-plate splines, and canonical variates analysis (CVA) analyses were performed. As expected, the shape of the greater sciatic notch provided the best separation between the sexes. Surprisingly, however, the shape of the orbits performed better that ramus flexure. Several possible explanations for this result are possible, which include the possibility that orbit shape is more sexually dimorphic than previously expected, or that biological reality is not reflected by this technique. More research is, however, needed. PMID- 16245345 TI - Bioleaching of sulfidic tailing samples with a novel, vacuum-positive pressure driven bioreactor. AB - This study presents a design for a novel bioreactor that uses alternating vacuum and positive pressure cycles to transfer acidic leach solution in and out of contact with finely ground sulfidic mine tailings. These tailings constitute an environmental problem that needs experimental data to support the development of management and control strategies. A conventional stirred tank bioreactor was used as a reference system. Both bioreactors were inoculated with mixed cultures of acidophilic iron and sulfur oxidizers. The rate of the bioleaching of tailings was 0.50 +/- 0.14 g Fe/L . day in the stirred tank bioreactor and 0.17 +/- 0.05 g Fe/L . day in the novel bioreactor. Microbial populations were identified in the two-bioreactor systems by analysis of 16S rRNA genes involving amplification, denaturing gradient gel electrophoresis (DGGE), cloning, and sequencing. The inoculum contained sulfur-oxidizing Acidithiobacillus caldus and Acidithiobacillus thiooxidans, iron oxidizers from the genera Leptospirillum and Ferroplasma, and a chemoorganotrophic Alicyclobacillus sp. During bioleaching of the tailings, the microbial populations in both bioreactors were similar to the inoculum culture, except that At. thiooxidans outgrew At. caldus. Sequences consistent with a Sulfobacillus sp. were amplified from both bioreactor samples although this bacterium was initially below the level of detection in the inoculum. After prolonged operation, Ferroplasma acidiphilum and an uncultured bacterium related to the CFB group were also detected in the novel bioreactor, whereas Sulfobacillus sp. was no longer detected. The novel bioreactor has potential uses in other areas of environmental biotechnology that involves periodic contact of liquids with solid substrates. PMID- 16245346 TI - Engineering of osteoinductive grafts by isolation and expansion of ovine bone marrow stromal cells directly on 3D ceramic scaffolds. AB - In this work, we investigated whether osteoinductive constructs can be generated by isolation and expansion of sheep bone marrow stromal cells (BMSC) directly within three-dimensional (3D) ceramic scaffolds, bypassing the typical phase of monolayer (2D) expansion prior to scaffold loading. Nucleated cells from sheep bone marrow aspirate were seeded into 3D ceramic scaffolds either by static loading or under perfusion flow and maintained in culture for up to 14 days. The resulting constructs were exposed to enzymatic treatment to assess the number and lineage of extracted cells, or implanted subcutaneously in nude mice to test their capacity to induce bone formation. As a control, BMSC expanded in monolayer for 14 days were also seeded into the scaffolds and implanted. BMSC could be isolated and expanded directly in the 3D ceramic scaffolds, although they proliferated slower than in 2D. Upon ectopic implantation, the resulting constructs formed a higher amount of bone tissue than constructs loaded with the same number of 2D-expanded cells. Constructs cultivated for 14 days generated significantly more bone tissue than those cultured for 3 days. No differences in bone formation were found between samples seeded by static loading or under perfusion. In conclusion, the culture of bone marrow nucleated cells directly on 3D ceramic scaffolds represents a promising approach to expand BMSC and streamline the engineering of osteoinductive grafts. PMID- 16245347 TI - Control of continuous polyhydroxybutyrate synthesis using calorimetry and flow cytometry. AB - The substrate-carbon flow can be controlled in continuous bioreactor cultures by the medium composition, for example, by the C/N ratio. The carbon distribution is optimal when a maximum fraction flows into the desired product and the residual is just sufficient to compensate for the dilution of the microbial catalyst. Undershooting of the latter condition is reflected immediately by changes in the Gibbs energy dissipation and cellular states. Two calorimetric measurement principles were applied to optimize the continuous synthesis of polyhydroxybutyrate (PHB) by Variovorax paradoxus DSM4065 during growth with constantly increasing supply rates of fructose or toxic phenol. Firstly, the changed slope of the heat production rate in a complete heat balanced bioreactor (CHB) indicated optimum carbon channeling into PHB. The extent of the alteration depended directly on the toxic properties of the substrate. Secondly, a flow through calorimeter was connected with the bioreactor as a "measurement loop." The optimum substrate carbon distribution was indicated by a sudden change in the heat production rate independent of substrate toxicity. The sudden change was explained mathematically and exploited for the long-term control of phenol conversion into PHB. LASER flow cytometry measurements distinguished between subpopulations with completely different PHB-content. Populations grown on fructose preserved a constant ratio of two subpopulations with double and quadruple sets of DNA. Cells grown on phenol comprised a third subpopulation with a single DNA set. Rising phenol concentrations caused this subpopulation to increase. It may thus be considered as an indicator of chemostress. PMID- 16245348 TI - Microbial synthesis of triacetic acid lactone. AB - Native g2ps1-encoded 2-pyrone synthase (2-PS) from Gerbera hybrida, a mutant Brevibacterium ammoniagenes fatty acid synthase B (FAS-B) and two different mutants of Penicillium patulum 6-methylsalycilic acid synthase (6-MSAS) are examined to identify the best enzyme to recruit for the microbial synthesis of triacetic acid lactone (TAL). To identify the best microbial host for these evaluations, the native TAL-synthesizing activity of g2ps1-encoded 2-PS is expressed in recombinant Escherichia coli and Saccharomyces cerevisiae constructs. Five-fold higher expression levels of 2-PS are observed in S. cerevisiae. Consequently, microbial synthesis of TAL focuses on S. cerevisiae constructs. Comparison of different promoters for the expression of g2ps1 in S. cerevisiae indicates that the alcohol dehydrogenase II promoter (P(ADH2)) affords the highest expression levels of 2-PS. As a result, the genes encoding the various TAL-synthesizing enzyme activities are expressed in S. cerevisiae from a P(ADH2) promoter. To extend TAL-synthesizing activity beyond g2ps1-encoded 2-PS, the ketoreductase domains of fasB-encoded FAS-B and 6-MSAS-encoded 6-MSAS are modified using a single mutation. Modification of the nicotinamide cofactor binding site of 6-MSAS with a triple mutation is also examined. Separate S. cerevisiae constructs expressing native g2ps1, mutant Y2226F fasB, mutant Y1572F 6-MSAS, and mutant G1419A-G1421P-G1424A 6-MSAS are cultured under the same fermentor-controlled conditions. The highest concentration (1.8 g/L) and yield (6%) of TAL are synthesized from glucose by S. cerevisiae expressing the Y1572F mutant of 6-MSAS. PMID- 16245349 TI - Thermostabilization of porcine kidney D-amino acid oxidase by a single amino acid substitution. AB - D-amino acid oxidase (DAO) is of considerable practical importance, such as bioconversion and enzymatic assay. In this study, we succeeded in obtaining a thermostable mutant DAO from porcine kidney by a single amino acid substitution. This mutant enzyme, F42C, was stable at 55 degrees C, while the wild-type enzyme was stable only up to 45 degrees C. The Km values of F42C for D-amino acids was about half of those of the wild-type enzyme. This mutant DAO with improved stability and affinity for its substrates is advantageous for the determination of D-amino acids. PMID- 16245350 TI - Compact reduced thioredoxin structure from the thermophilic bacteria Thermus thermophilus. AB - The X-ray crystallographic structure of a thioredoxin from Thermus thermophilus was solved to 1.8 A resolution by molecular replacement. The crystals' space group was C2 with cell dimensions of a = 40.91, b = 95.44, c = 56.68 A, beta =91.41 degrees, with two molecules in the asymmetric unit. Unlike the reported thioredoxin structures, the biological unit of T. thermophilus thioredoxin is a dimer both in solution and in the crystal. The fold conforms to the "thioredoxin fold" that is common over a class of nine protein families including thioredoxin; however, the folded portion of this protein is much more compact than other thioredoxins previously solved by X-ray crystallography being reduced by one alpha-helix and one beta-strand. As with other thioredoxins, the active site is highly conserved even though the variation in sequence can be quite large. The T. thermophilus thioredoxin has some variability at the active site, especially compared with previously solved structures from bacterial sources. PMID- 16245351 TI - Exploitation of the alcohol dehydrogenase-acetone NADP-regeneration system for the enzymatic preparative-scale production of 12-ketochenodeoxycholic acid. AB - The performance of a new NADP-regeneration system, based on the use of alcohol dehydrogenase (ADH)-acetone, has been investigated for the regioselective oxidation of cholic acid (1) to 12-ketochenodeoxycholic acid (2). Enzymes stabilities and substrate and/or product inhibitory effects under defined synthetic reaction conditions have been evaluated. The optimized system, based on a 4% w/v solution of 1 in a reaction mixture containing 25% v/v acetone, allowed the preparative scale transformation of 1 into 2 with a 92% conversion. PMID- 16245352 TI - Reducing adverse drug events in the outpatient chemotherapy setting: attention must be paid. PMID- 16245353 TI - Medication safety in the ambulatory chemotherapy setting. AB - BACKGROUND: Little is known concerning the safety of the outpatient chemotherapy process. In the current study, the authors sought to identify medication error and potential adverse drug event (ADE) rates in the outpatient chemotherapy setting. METHODS: A prospective cohort study of two adult and one pediatric outpatient chemotherapy infusion units at one cancer institute was performed, involving the review of orders for patients receiving medication and/or chemotherapy and chart reviews. The adult infusion units used a computerized order entry writing system, whereas the pediatric infusion unit used handwritten orders. Data were collected between March and December 2000. RESULTS: The authors reviewed 10,112 medication orders (8008 adult unit orders and 2104 pediatric unit orders) from 1606 patients (1380 adults and 226 pediatric patients). The medication error rate was 3% (306 of 10,112 orders). Of these errors, 82% occurring in adults (203 of 249 orders) had the potential for harm and were potential ADEs, compared with 60% of orders occurring in pediatric patients (34 of 57 orders). Among these, approximately one-third were potentially serious. Pharmacists and nurses intercepted 45% of potential ADEs before they reached the patient. Several changes were implemented in the adult and pediatric settings as a result of these findings. CONCLUSIONS: In the current study, the authors found an ambulatory medication error rate of 3%, including 2% of orders with the potential to cause harm. Although these rates are relatively low, there is clearly the potential for serious patient harm. The current study identified strategies for prevention. PMID- 16245354 TI - Twelve-year mortality results of a randomized trial of 2 versus 5 years of adjuvant tamoxifen for postmenopausal early-stage breast carcinoma patients (SITAM 01). AB - BACKGROUND: This study evaluated the impact on overall survival (OS) of 2 versus 5 years adjuvant tamoxifen in early breast carcinoma patients after 12 years of follow-up. METHODS: Women with breast carcinoma T1-3, N0-3, M0, aged 50-70 years, were eligible for this multicenter randomized Phase III trial. Patients event free after 2 years of tamoxifen therapy (TAM) were randomly assigned to stop or continue TAM (20 mg/day) for an additional 3 years. The primary endpoint was disease-free survival. Secondary endpoints included OS and toxicity. RESULTS: From 1989 through 1996, 1901 patients were randomly assigned either to stop treatment (n = 958) or to continue TAM (n = 943). Overall, 98% of patients alive at the previous report (n = 1611) had updated information about OS, of whom 549 had died. The median duration of postrandomization follow-up was 115 months (interquartile range, 86-137). No statistically significant differences between the two arms were detected in the whole population (hazard ratio [HR], 1.02; 95% confidence interval [CI], 0.86-1.22) and in estrogen receptor (ER)-positive patients (HR, 0.90; 95% CI, 0.72-1.13). In the latter group, survival curves started to diverge after 90 months, showing a trend in favor of the 5-year arm. In younger (age < or =55 yrs) ER-positive patients longer TAM was associated with a 44% decrease in the risk of death (HR, 0.56; 95% CI, 0.31-1.00), while no clear benefit was documented in women older than 55 years of age (HR, 0.98; 95% CI, 0.77-1.25). CONCLUSIONS: The benefits of longer TAM on OS start to emerge only after 9 years from diagnosis and seem to be more relevant in younger ER-positive women. PMID- 16245356 TI - Expression of stathmin family genes in the murine uterus during early pregnancy. AB - Stathmin, a cytosolic phosphoprotein that regulates microtubule dynamics during cell-cycle progression, is abundantly expressed at embryo implantation sites in rats. Here, we characterized the expression of stathmin and its family genes in the murine uterus during the peri-implantation period. Stathmin protein was expressed in the glandular and luminal epithelium, blood vessels, and stromal cells on day 3 of pregnancy. On the day of implantation (day 5), stathmin was mainly localized in blood vessels in the endometrium. On day 7, intense stathmin expression was limited to capillary vessels and secondary decidual cells. Stathmin expression was higher at implantation sites than at uterine segments between implantation sites and increased during oil-induced decidualization. Although the artificially-induced deciduoma weights and number of implantation sites were similar between stathmin-knockout (KO) and wild-type (WT) mice, the stathmin-KO mice had fewer newborn pups (reduced by 30%). The expression of alkaline phosphatase, desmin, and cyclin D3 was attenuated in decidual zones of stathmin-KO mice. Messenger RNA level of the stathmin family gene, SCG10, was high at the time of decidualization in WT and stathmin-KO mice. In contrast, the others of stathmin family members, SCLIP and RB3 were highly expressed in stathmin-KO mice compared to WT mice. These results suggest that stathmin and stathmin family genes are expressed in the murine endometrium with enhanced expression in the implantation or the decidualization process. PMID- 16245355 TI - Gradual DNA demethylation of the Oct4 promoter in cloned mouse embryos. AB - During differentiation, somatic cell nuclei acquire unique patterns of epigenetic modifications, such as DNA methylation, which affect the transcriptional activity of specific genes. Upon transfer into oocytes, however, the somatic nucleus undergoes reprogramming of these epigenetic modifications to achieve pluripotency. Oct4 is one of the critical pluripotency regulators, and is expressed in the germ line, including the pluripotent early embryonic cells. Previous studies showed that the upstream regulatory sequences of the Oct4 gene are distinctly methylated in somatic cells, and the DNA methylation of the regulatory sequences suppresses the transcriptional activity. Thus, successful reprogramming of the somatic cell nucleus to gain pluripotency must be accompanied by the demethylation of the Oct4 regulatory sequences. Here, we investigated the methylation pattern of the Oct4 promoter during early development of cloned mouse embryos. We found that the Oct4 promoter was only gradually demethylated during the early cleavage stages and that the ineffective demethylation of the promoter was associated with developmental retardation. We also found that the upstream sequences of the other pluripotency regulators, namely Nanog, Sox2, and Foxd3, were considerably under-methylated in cumulus cells. These results suggest that the Oct4 gene, as compared to the other pluripotency regulators, needs to undergo extensive demethylation during nuclear reprogramming, and that the failure of such demethylation is associated with inefficient development of cloned somatic cell embryos. PMID- 16245357 TI - Transmission of mitochondrial DNA in pigs and progeny derived from nuclear transfer of Meishan pig fibroblast cells. AB - In embryos derived by nuclear transfer (NT), fusion, or injection of donor cells with recipient oocytes caused mitochondrial heteroplasmy. Previous studies have reported varying patterns of mitochondrial DNA (mtDNA) transmission in cloned calves. Here, we examined the transmission of mtDNA from NT pigs to their progeny. NT pigs were created by microinjection of Meishan pig fetal fibroblast nuclei into enucleated oocytes (maternal Landrace background). Transmission of donor cell (Meishan) mtDNA was analyzed using 4 NT pigs and 25 of their progeny by PCR-mediated single-strand conformation polymorphism (PCR-SSCP) analysis, PCR RFLP, and a specific PCR to detect Meishan mtDNA single nucleotide polymorphisms (SNP-PCR). In the blood and hair root of NT pigs, donor mtDNAs were not detected by PCR-SSCP and PCR-RFLP, but detected by SNP-PCR. These results indicated that donor mtDNAs comprised between 0.1% and 1% of total mtDNA. Only one of the progeny exhibited heteroplasmy with donor cell mtDNA populations, ranging from 0% to 44% in selected tissues. Additionally, other progeny of the same heteroplasmic founder pig were analyzed, and 89% (16/18) harbored donor cell mtDNA populations. The proportion of donor mtDNA was significantly higher in liver (12.9 +/- 8.3%) than in spleen (5.0 +/- 3.9%), ear (6.7 +/- 5.3%), and blood (5.8 +/- 3.7%) (P < 0.01). These results demonstrated that donor mtDNAs in NT pigs could be transmitted to progeny. Moreover, once heteroplasmy was transmitted to progeny of NT-derived pigs, it appears that the introduced mitochondrial populations become fixed and maternally-derived heteroplasmy was more readily maintained in subsequent generations. PMID- 16245358 TI - Comparison of melatonin, vitamin E and L-carnitine in the treatment of neuro- and hepatotoxicity induced by thioacetamide. AB - This study was designed to evaluate and compare the effect of melatonin, vitamin E and L-carnitine on brain and liver oxidative stress and liver damage. Oxidative stress and hepatic failure were produced by a single dose of thioacetamide (TAA) (150 mg kg(-1)) in Wistar rats. A dose of either melatonin (3 mg kg(-1)) vitamin E (20 mg kg(-1) ) or L-carnitine (100 mg kg(-1)) was used. Blood samples were taken from the neck vasculature in order to determine ammonium, blood urea nitrogen (BUN) and liver enzymes. Lipid peroxidation products, glutathione (GSH) content and antioxidative enzymes were determined in cerebral and hepatic homogenates. The results showed a decrease in BUN and in the antioxidant enzymes activities and GSH in the brain and liver. Likewise, TAA induced significant enhancement of lipid peroxidation products levels in both liver and brain, as well as in ammonia values. Melatonin, vitamin E and L-carnitine, although melatonin more significantly, decreased the intensity of the changes produced by the administration of TAA alone. Furthermore melatonin combined with TAA, decreased the ammonia levels and increased the BUN values compared with TAA animals. Also it was more effective than vitamin E or L-carnitine in these actions. These data show the protective effect of these agents, especially melatonin, against oxidative stress and hepatic damage present in fulminant hepatic failure. PMID- 16245359 TI - Snake venom hyaluronidase: a therapeutic target. AB - The diffusion of toxins from the site of a bite into the circulation is essential for successful envenomation. Degradation of hyaluronic acid in the extracellular matrix (ECM) by venom hyaluronidase is a key factor in this diffusion. Hyaluronidase not only increases the potency of other toxins but also damages the local tissue. In spite of its important role, little attention has been paid to this enzyme. Hyaluronidase exists in various isoforms and generates a wide range of hyaluronic acid degradation products. This suggests that beyond its role as a spreading factor venom hyaluronidase deserves to be explored as a possible therapeutic target for inhibiting the systemic distribution of venom and also for minimizing local tissue destruction at the site of the bite. PMID- 16245360 TI - Development of a novel Ag-specific immunotherapy using CpG oligodeoxynucleotides in a new, unique mouse cutaneous eosinophilic inflammation model. AB - The number of patients with severe atopic dermatitis (AD) has been on the rise recently. We are therefore urgently in need of a treatment that can suppress Th2 cell-mediated responses in an Ag-specific fashion. Oligodeoxynucleotides (ODN)containing CpG motifs (CpG ODN) have been highlighted as immunomodulators that reduce Th2-mediated responses. To determine the effect of CpG ODN on Th2 mediated skin inflammation, we first developed a reproducible murine model of protein Ag-induced eosinophilic inflammation that is accompanied by epidermal acanthosis and increased serum IgE levels as seen in AD. In this model we found that treatment with CpG ODN during epicutaneous sensitization in previously i.p. primed mice prevented the development of Th2-mediated responses. Furthermore, to evaluate the therapeutic effect of CpG ODN on established eosinophilic inflammation, mice were treated with a course of the immunotherapy at a skin site remote from the area of Ag application prior to the second 1-wk epicutaneous exposure to Ag. Therapeutic treatment with CpG ODN plus Ag, but not that with CpG ODN alone, could reverse the established eosinophilic inflammation. The presented results provide strong evidence for the feasibility of a novel Ag-specific immunomodulator to treat cutaneous eosinophilic inflammation such as that characteristically found in patients with severe AD. PMID- 16245361 TI - Altered primary CD8+ T cell response to a modified virus Ankara(MVA)-vectored vaccine in the absence of CD4+ T cell help. AB - T cell receptor-transgenic F5 mice were used to assess primary CD8+ T cell responses to a modified virus Ankara (MVA)-vectored vaccine in the absence of CD4+ T cell help. Naive, CD8-enriched, CFSE-labelled F5 cells were transferred into normal or CD4+ cell-depleted mice and the mice were vaccinated with MVA.HIVA NP. At different time points during the primary response, F5 cells were re isolated and analysed on divisional basis for a number of parameters. We demonstrated that the primary CD8+ T cell response in the absence of CD4+ T cell help differed from that in normal CD4+ cell-undepleted mice. While in the absence of CD4+ T cell help, the initial migratory progress from the local response to a systemic one was not grossly affected, the proportion of dying F5 cells during the expansion phase was markedly increased and resulted in an overall smaller expansion and significantly decreased frequency of CD8+ T cell memory after contraction. T cells primed without help displayed accelerated proliferation and activation, while expression of interferon-gamma remained similar. These phenomena were observed in the lymph nodes draining the MVA.HIVA-NP immunization site and were similar, but delayed by 2-3 days in spleen and non-draining lymph nodes. PMID- 16245362 TI - The heat shock protein Hsp70 enhances antigen-specific proliferation of human CD4+ memory T cells. AB - Heat shock proteins (HSP) can interact with a wide variety of peptides and the resulting HSP:peptide complexes are known to be highly immunogenic. The ability of HSP:peptide complexes to elicit CD8+ T cell responses by cross-presentation of exogenous antigen via MHC class I is well known. In contrast, their role in the activation of CD4+ T cells is less clearly defined, although several recent studies in mice and T cell lines suggest an involvement of HSP in the presentation of antigenic peptides via MHC class II. In this study we have investigated the potential of antigenic peptides from tetanus toxin and influenza hemagglutinin complexed to the human stress-inducible Hsp70 to enhance activation and proliferation of human memory CD4+ T cells. Hsp70:peptide complexes were found to amplify the proliferation of antigen-specific CD4+ T cells as confirmed by HLA-DR tetramer staining. Complex formation of the antigenic peptide with Hsp70 was absolutely required to elicit an antigen-specific amplification. This effect was most pronounced at low doses of antigen and decreasing APC/CD4+ T cell ratios. Taken together, we show the potential of Hsp70 to enhance antigen specific CD4+ T cell proliferation and to increase the immunogenicity of presented peptides in human CD4+ T cells. PMID- 16245363 TI - Facile synthesis of phosphonamidate- and phosphonate-linked phosphonopeptides. AB - A direct method for the preparation of phosphonamidate- and phosphonate-linked phosphonopeptides has been developed. Using this method, both phosphonopeptides were prepared in acceptable yields directly from simple and commercially available chemicals in one-pot reactions of benzyl carbamate, aldehydes, and methyl dichlorophosphite, followed by aminolysis with amino acid esters or alcoholysis with hydroxy esters. PMID- 16245365 TI - Optimized lipopolyplex formulations for gene transfer to human colon carcinoma cells under in vitro conditions. AB - BACKGROUND: Polycation (PC, polyplex), cationic lipid (CL, lipoplex), and a combination of PC/CL (lipopolyplex) formulations were investigated for gene transfer to slow-proliferating human colon carcinoma cell lines (COGA). METHODS: The luciferase reporter gene was complexed with either PC, CL, or PC/CL. PCs included linear (PEI22lin, 22 kDa) and branched polyethylenimine (PEI2k, 2 kDa; PEI25br, 25 kDa) and poly-L-lysine (PLL18 with 18 lysine monomers). CLs included DOCSPER, DOSPER and DOTAP. Lipopolyplexes were formed by either sequentially first mixing DNA with PC or CL, followed by addition of CL or PC, respectively, or simultaneously with both PC and CL. Particle size and zeta-potential were determined and gene transfer and cytotoxicity were quantified on COGA-3, -5, -12, HeLa and Sw480 cells. RESULTS: The highest gene transfer was achieved when DNA was first complexed with PC followed by CL. At low ionic strength, particles were small (50-130 nm) with a zeta-potential of +20-40 mV. At physiological ionic strength, only lipoplexes of DOCSPER or DOSPER and their respective lipopolyplexes with PEI25br were stable to aggregation (140-220 nm). Lipopolyplexes of PEI25br were between 5- to 400-fold more efficient compared to the corresponding lipoplexes or polyplexes in all cases. Chloroquine did not significantly affect lipopolyplex-mediated gene transfer. CONCLUSIONS: Lipopolyplex formulations of PEI25br in combination with multivalent CLs (DOCSPER, DOSPER) are promising tools for in vitro and potentially also in vivo gene transfer to colorectal cancer cells. PMID- 16245367 TI - Hepatitis B vaccination and multiple sclerosis: the jury is still out. PMID- 16245368 TI - Molecular basis for regulation of Src by the docking protein p130Cas. AB - The docking protein p130Cas (Cas) becomes tyrosine-phosphorylated in its central substrate domain in response to extracellular stimuli such as integrin-mediated cell adhesion, and transmits signals through interactions with various intracellular signaling molecules such as the adaptor protein Crk. Src-family kinases (SFKs) bind a specific site in the carboxyl-terminal region of Cas and subsequently SFKs phosphorylate progressively the substrate domain in Cas. In this study crystallography, mutagenesis and binding assays were used to understand the molecular basis for Cas interactions with SFKs. Tyrosine phosphorylation regulates binding of Cas to SFKs, and the primary site for this phosphorylation, Y762, has been proposed. A phosphorylated peptide corresponding to Cas residues 759MEDpYDYVHL767 containing the key phosphotyrosine was crystallized in complex with the SH3-SH2 domain of the SFK Lck. The results provide the first structural data for this protein-protein interaction. The motif in Cas 762pYDYV binds to the SH2 domain in a mode that mimics high-affinity ligands, involving dual contacts of Y762 and V765 with conserved residues in SFK SH2 domains. In addition, Y764 is in position to make an electrostatic contact after phosphorylation with a conserved SFK arginine that mediates interactions with other high-affinity SH2 binders. These new molecular data suggest that Cas may regulate activity of Src as a competing ligand to displace intramolecular interactions that occur in SFKs (between the C-terminal tail and the SH2 domain) and restrain and down-regulate the kinase in an inactive form. PMID- 16245369 TI - About histone H1 phosphorylation during mitosis. PMID- 16245370 TI - Ischemic preconditioning preserves proton leakage from mitochondrial membranes but not oxidative phosphorylation during heart reperfusion. AB - The aim of this study was to evaluate the role of mitochondria in the recovery of cardiac energetics induced by ischaemic preconditioning at reperfusion. Isolated rat hearts were aerobically perfused (control), subjected to global ischaemia and reperfusion (reperfusion), or subjected to 3 brief cycles of ischaemia/reperfusion and then to the protocol of reperfusion (preconditioning). At the end of the perfusion, antimycin A was delivered to the heart for 25 min, to inhibit mitochondrial respiration and stimulate glycolysis. The increased amount of lactate released in the coronary effluent was correlated with the number of viable cells producing this end-product of glycolysis. Preconditioned hearts released 18% more lactate than reperfused hearts (p < 0.05). This result indicates that preconditioning partially preserved cell viability, as was also evidenced by the MTT assay performed on cardiac biopsies. The difference between antimycin A-stimulated and basal lactate concentration, representing the contribution of mitochondria to the overall energetics of cardiac tissue, was also 18% more elevated in the preconditioned hearts than in the reperfused hearts (p < 0.01). The study of the respiratory function of mitochondria isolated at the end of perfusion, showed that preconditioning did not improve the oxygen dependent production of ATP (state 3 respiration, ADP/O). On the contrary, state 4 respiration, which is related to proton leakage, was 35.0% lower in the preconditioned group than reperfusion group (p < 0.05). Thus, preconditioning ameliorates cardiac energetics by preserving cell death, but without affecting mitochondrial oxidative phosphorylation. Mitochondria can contribute to cell survival by the attenuation of proton leak from inner membrane. PMID- 16245371 TI - Maternal serum biochemistry at 11-13(+6) weeks in relation to the presence or absence of the fetal nasal bone on ultrasonography in chromosomally abnormal fetuses: an updated analysis of integrated ultrasound and biochemical screening. AB - BACKGROUND: Screening for trisomy 21 by a combination of maternal age, fetal nuchal translucency (NT) thickness and maternal serum free beta-hCG and pregnancy associated plasma protein-A (PAPP-A) at 11-13(+6) weeks of gestation is associated with a detection rate of 90%, for a false-positive rate of 5%. Recent evidence suggests that in about 70% of fetuses with trisomy 21 the nasal bone is not visible at the 11-13(+6) week scan and that the frequency of absence of nasal bone differs in different ethnic groups. In addition, there is a relationship between absent nasal bone and nuchal translucency thickness. In a preliminary study we showed that while PAPP-A levels were lower and free beta-hCG levels were higher in trisomy 21 fetuses with an absent nasal bone, this difference was not statistically different. In fetuses with trisomy 13 and trisomy 18, there is also a high (57 and 67%) incidence of an absent nasal bone. The aim of this present study was to extend our examination of whether the level of maternal serum biochemical markers is independent of the presence or absence of the nasal bone in cases with trisomy 21 and to ascertain if any differences exist in cases with trisomies 13 and 18. METHODS: This study data comprised 100 trisomy 21 singleton pregnancies at 11-13(+6) weeks of gestation from our previous study and an additional 42 cases analysed as part of routine OSCAR screening. A total of 34 cases with trisomy 18 and 12 cases with trisomy 13 were also available. Ultrasound examination was carried out for measurement of fetal NT and assessment of the presence or absence of the fetal nasal bone. Maternal serum free beta-hCG and PAPP-A were measured using the Kryptor rapid random access immunoassay analyser (Brahms Diagnostica AG, Berlin). The distribution of maternal serum free beta-hCG and PAPP-A in chromosomally abnormal fetuses with absent and present nasal bone was examined. RESULTS: The nasal bone was absent in 29 and present in 13 of the new trisomy 21 cases and in 98 (69%) and 44 respectively in the combined series. For the trisomy 18 cases, the nasal bone was absent in 19 (55.9%) cases and in 3 (25%) of cases of trisomy 13. There were no significant differences in median maternal age, median gestational age, NT delta, free beta hCG MoM and PAPP-A MoM in trisomy 21 fetuses with and without a visible nasal bone, and similarly for those with trisomies 13 or 18. For a false-positive rate of 5%, it was estimated that screening with the four markers in combination with maternal age would be associated with a detection rate of 96% of cases with trisomy 21. For a false-positive rate of 0.5%, the detection rate was 88%. CONCLUSIONS: There is no relationship between an absent fetal nasal bone and the levels of maternal serum PAPP-A or free beta-hCG in cases with trisomies 13, 18 or 21. An integrated sonographic and biochemical test at 11-13(+6) weeks can potentially identify about 88% of trisomy 21 fetuses for a false-positive rate of 0.5%. PMID- 16245372 TI - Effects of intraperitoneally administered ubiquinone on the level of total lipid and fatty acids in rat liver. AB - The purpose of the present study was to evaluate the effect of ubiquinone (coenzyme Q-10) on total lipid and fatty acid composition of liver tissues in rats. Twenty male wistar rats were randomly divided into two groups. The first group was used as a control. The second group received ubiquinone (8 mg/every other day) intraperitoneally. This administration was done for a period of 38 days. Body weight increases in animals fed diets for 38 days were on average 35 g in control group (C), and only 11 g in the ubiquinone group. Total lipid content of liver tissues in the ubiquinone group (UB) decreased significantly (p < 0.0001) compared to the control group (C). The ratio of 22:6 and total omega3 fatty acid in the UB increased (p < 0.01) compared to C. While the level of oleic acid (18:1), palmitoleic acid (16:1) and total monounsaturated fatty acid (MUFA) in UB significantly decreased (p < 0.01, p < 0.001, p < 0.001, respectively), the level of stearic acid (18:0) in liver tissue increased (p < 0.05) in the same group when compared to C. Stearoyl-CoA desaturase (SCD) is the rate-limiting enzyme catalyzing the synthesis of monounsaturated fatty acid mainly oleate (18:1). We speculate that ubiquinone inhibits SCD activity. SCD is an important metabolic control point in body weight regulation. Our results indicate that ubiquinone supplementation may have an inhibitory effect on obesity and it seems that the level of 22:6 in liver increased due to ubiquinone. PMID- 16245373 TI - Binding of helix-threading peptides to E. coli 16S ribosomal RNA and inhibition of the S15-16S complex. AB - Helix-threading peptides (HTPs) constitute a new class of small molecules that bind selectively to duplex RNA structures adjacent to helix defects and project peptide functionality into the dissimilar duplex grooves. To further explore and develop the capabilities of the HTP design for binding RNA selectively, we identified helix 22 of the prokaryotic ribosomal RNA 16S as a target. This helix is a component of the binding site for the ribosomal protein S15. In addition, the S15-16S RNA interaction is important for the ordered assembly of the bacterial ribosome. Here we present the synthesis and characterization of helix threading peptides that bind selectively to helix 22 of E. coli 16S RNA. These compounds bind helix 22 by threading intercalation placing the N termini in the minor groove and the C termini in the major groove. Binding is dependent on the presence of a highly conserved purine-rich internal loop in the RNA, whereas removal of the loop minimally affects binding of the classical intercalators ethidium bromide and methidiumpropyl-EDTAFe (MPEFe). Moreover, binding selectivity translates into selective inhibition of formation of the S15-16S complex. PMID- 16245374 TI - Unidirectional threading synthesis of isomer-free [2]rotaxanes. AB - The threading of an alpha-cyclodextrin (alpha-CyD) by an unsymmetrical dumbbell generally results in two isomeric [2]rotaxanes differing in the orientation of the alpha-CyD. In this work, two methods have been developed for the unidirectionally threading an alpha-CyD to obtain isomer-free [2]rotaxanes. These methods use the Suzuki coupling of a boronic acid derivative and a halide in aqueous alkaline solution. The conformations of the two unidirectional [2]rotaxanes-R3 and R4 were determined by 2D 1H ROESY NMR spectra. The optical spectral studies revealed that each of the two [2]rotaxanes can proceed with E/Z photoisomerization and shuttling motions of the alpha-CyD ring on the thread under alternating irradiation at 330 and 275 nm, accompanied by fluorescence intensity changes at 530 nm. The induced circular dichroism (ICD) spectra of another two analogous [2]rotaxanes R1 and R2 were also studied. Distinctive ICD signal changes resulting from the photoisomerization with respect to the movements of alpha-CyD were detected. This demonstrates that, besides the fluorescence, ICD signal is another way to identify the shuttling motions of alpha-CyD in these [2]rotaxanes. PMID- 16245375 TI - Oligoethylene chains terminated by ferrocenyl end groups: synthesis, structural properties, and two-dimensional self-assembly on surfaces. AB - A series of unsaturated long-chain-bridged diferrocenes Fc-(CH2)n-CH=CH-(CH2)n-Fc (4 a-e) was synthesized by means of olefin metathesis. Subsequent catalytic hydrogenation furnished the saturated alpha,omega-bis-ferrrocenyl oligoethylene products Fc-(CH2)m-Fc (5). Members of both series formed highly ordered laminar structures at the highly oriented pyrolytic graphite (HOPG) solid/liquid interface or on the Ag(110) surface, which were characterized by STM. Details of the structural features of these ordered physisorbed surface assemblies of 4 and 5 were analyzed by comparison with DFT calculations on model systems and with the characteristic packing modes of these systems in the crystal. PMID- 16245376 TI - Synthesis of heterobimetallic Ru-Mn complexes and the coupling reactions of epoxides with carbon dioxide catalyzed by these complexes. AB - The heterobimetallic complexes [(eta5-C5H5)Ru(CO)(mu-dppm)Mn(CO)4] and [(eta5 C5Me5)Ru(mu-dppm)(mu-CO)2Mn(CO)3] (dppm = bis-diphenylphosphinomethane) have been prepared by reacting the hydridic complexes [(eta5-C5H5)Ru(dppm)H] and [(eta5 C5Me5)Ru(dppm)H], respectively, with the protonic [HMn(CO)5] complex. The bimetallic complexes can also be synthesized through metathetical reactions between [(eta5-C5R5)Ru(dppm)Cl] (R = H or Me) and Li+[Mn(CO)5]-. Although the complexes fail to catalyze the hydrogenation of CO2 to formic acid, they catalyze the coupling reactions of epoxides with carbon dioxide to yield cyclic carbonates. Two possible reaction pathways for the coupling reactions have been proposed. Both routes begin with heterolytic cleavage of the RuMn bond and coordination of an epoxide molecule to the Lewis acidic ruthenium center. In Route I, the Lewis basic manganese center activates the CO2 by forming the metallocarboxylate anion which then ring-opens the epoxide; subsequent ring closure gives the cyclic carbonate. In Route II, the nucleophilic manganese center ring-opens the ruthenium-attached epoxide to afford an alkoxide intermediate; CO2 insertion into the RuO bond followed by ring-closure yields the product. Density functional calculations at the B3LYP level of theory were carried out to understand the structural and energetic aspects of the two possible reaction pathways. The results of the calculations indicate that Route II is favored over Route I. PMID- 16245377 TI - Thermodynamic stability and ultrasmall-size effect of nanodiamonds. PMID- 16245378 TI - A high-throughput investigation of the role of pH, temperature, concentration, and time on the synthesis of hybrid inorganic-organic materials. PMID- 16245379 TI - Kinetic resolution of oxazinones: an organocatalytic approach to enantiomerically pure beta-amino acids. PMID- 16245380 TI - Anion-induced synthesis and combinatorial selection of polypyrrolic macrocycles. PMID- 16245381 TI - Direct readout of protein-protein interactions by mass spectrometry from protein DNA microarrays. PMID- 16245382 TI - Fibrous red phosphorus. PMID- 16245383 TI - [Ecological risk of antibiotic use: consequences for new molecule evaluation]. PMID- 16245384 TI - [How should potential selection for bacterial antibiotic resistance be defined?]. AB - Evolution of bacterial resistance to antibiotics could be better controlled if we knew how to define the potential of evolution of bacterial resistance towards a given antibiotic, prospectively. No unique method allows to do it. Pieces of answers can be obtained by using diverse methods using bacteriological, ecological pharmaco-epidemiological techniques and experimental models. Association of the results obtained with these various techniques is mandatory to understand the evolution potential of resistance towards a given antibiotic when it is used in the clinical setting. PMID- 16245385 TI - [Impact of population exposure on the risk of bacterial resistance]. AB - Exposure of populations to antibiotics is a condition for the emergence and the diffusion of bacteria resistant to antibiotics. Analysis of epidemiological causality between antibiotic exposure and expansion of antibiotic resistance in human pathogens requires the study of the basis for the emergence and the subsequent epidemic spread of the strains. Emergence of antibiotic resistance under the selective pressure of antibiotics is unavoidable. However, the rapidity of the emergence depends on several parameters including the biochemical mechanism and the genetic support of the resistance and on the couple bacteria/antibiotic considered. Several of these parameters may be studied and measured in vitro. In vivo, relationship between antibiotic exposure and risk for infection or colonisation by pneumococci resistant to beta-lactams or macrolides and methicillin-resistant staphylococci are among the best studied models. In particular for pneumococci, quantitative relationship between antibiotic consumption and resistance, including geographical correlations have been shown but do not allow to establish any causality relationship. Overall, there is no doubt on the question of the impact of antibiotic exposure and the risk of antibiotic resistance. However, the quantification of the risks remains to be accurately studied. PMID- 16245386 TI - [Link between resistance and failure in community-acquired respiratory infections]. AB - Resistance is one of failure's reasons. We tried, through clinical experience, to approach the magnitude and nature of the links, between phenotypically defined acquired resistance and clinical failure, in community acquired respiratory infections. An efficient resistance mecanism, able to suppress antibiotic action, is clearely associated to a risk of clinical failure (e.g. betalactamase secretion, target modification using methilation for macrolides, target mutation for fluoroquinoles). Resistance mecanism due to reduction of target affinity (pneumococcus and betalactams) progressively decreasing beta lactam activity depending on its expression, is at present time, not clearely associeted with clinical failure. Critical concentration, defining phenotypical resistance, is predictive of failure if it identifies a bacterial population owning an efficient resistance mecanism. It will not be predictive of failure if that concentration do not detect the resistance mecanism (e.g. parC mutation and levofloxacin) or if the link between antibiotic and resistant bacteria is not binary but depends also on pharmacokinetic parameters (pneumococcus and betalactam). Using resistance as a parametre for antibiotic choice, must integrate several elements: presence or not of a resistance mecanism, type and efficiency of the mecanism, links with clinical failure and antibiotic concentration, type and site of infection. Critical concentration is not allways the magic number that predict failure or success. PMID- 16245388 TI - Reply to letter regarding interpretation of results of ACTIVE Study. PMID- 16245387 TI - [Need for alternative approaches in the evaluation of new antibacterial antibiotics]. AB - The golden age of antibacterial antibiotics extend from year 1941 to the 1990s decade. At that time, something like an earth quake occurred: from the thirty molecules or so whose development was being achieved or was already marketed, only three were put on the French market, and faced the greatest difficulties to be prescribed by practicians, because: the knights of good practice want a strict limitation of their use to precise indications; the pharmaceutical companies find that the return on investment is almost impossible; the prescribers are stunned by the inconsistency between the MAs, the advances in science and the health economic authorities advices which claim that these products are not very interesting; the research for new antibiotics is stalling; thus, for the first time in 60 years, an iconoclastic question arises: do we need new antibiotics? However, while the debate is raging, many of us think "yes we do", as it is a duty to anticipate today the consequences of tomorrow's bacterial resistances. This paper presents three types of propositions to optimise the development of future molecules: sharpening of the data concerning preclinical security for a better predicting of both the activity and the toxicity; improvement in performances and organization of clinical trials, which implicates to reconsider some of the present methodological rules; inclusion in the evaluation data of some relevant and new features measuring the anti-bacterial activity while taking into account the present and future bacterial resistances. The development of new concepts to develop new drugs which would be active against tomorrow's bacteria compels us to manage in a new fashion today's systems, which have reached their own limits. PMID- 16245389 TI - Team-based occupational stress reduction: a European overview from the perspective of the OSCAR Project. AB - Work-related stress is a significant impediment to job satisfaction and healthy psycho-social functioning. It can alter the behaviour of the person involved and impair the quality of their life. In the European Union (EU), over the last decade, work-related stress has been consistently identified as one of the major workplace concerns--a challenge not only to the health of working people but also to the healthiness of their organizations. The study reported below attempted to address the issue of work-related stress through whole team training programmes, on a background of largely ineffective stress reduction training programmes offered to individuals within the workplace. This EU 'framework 5 Quality of Life' project focused instead on tackling the organizational level through training of mental health teams in five countries. The findings have significant implications to the conceptual, methodological and everyday organizational practice levels of tackling this central issue to the health of the workplace. PMID- 16245391 TI - Managing and promoting change: implementing the Leg Club model. AB - Patient knowledge of their disease may increase concordance to compression therapy. The Lindsay Leg Club model (Lindsay, 2004) empowers the community nursing team to truly get to know their communities and deliver local, high quality, evidence-based care for patients with leg ulcers and leg related problems. The author's experience of opening a Leg Club has lead to changes in clinical practice: re-evaluation of the use of long stretch bandages in favour of short stretch; replacing conventional compression bandage systems with two piece graduated compression hosiery; increased patient concordance post healing with increased use of compression hosiery; the introduction of toe bandaging and appropriate long-term management for those patients with lymphoedema; and developing the role of the nursing auxiliary in leg ulcer care. PMID- 16245390 TI - Community nurse use of Doppler ultrasound in leg ulcer assessment. AB - Doppler ultrasound is used by community nurses to measure the ankle brachial pressure index (ABPI). This is required before applying compression therapy for patients with chronic venous insufficiency and venous leg ulcers.However, emphasis on the ABPI result has resulted in inappropriate referrals to the vascular department which led the author to survey current practice within one primary care trust. Results illustrated variations in how nurses obtain training and maintain their competency in using Doppler ultrasound. This has an impact on the accuracy of interpretation of the ABPImeasurement and subsequent management of the patient. Practical issues also explained the difficulties nurses encountered in using the correct procedure within the community, which may result in unreliable measurements. There is a need to standardize training for all community nurses, and to review the structure of current clinical guidelines to enable a wider analysis of arterial assessment, in order to reduce the emphasis purely on the ABPI measurement. PMID- 16245392 TI - From costly treatment to cost-effective prevention: using Waterlow. AB - The Waterlow pressure ulcer risk assessment system has been in use for 20 years. In this article, Judy Waterlow describes how it can be used in the community, discusses the recent update to the system and explains how pressure ulcer risk assessments should be conducted. PMID- 16245394 TI - Worries over lab animals decision. PMID- 16245393 TI - Catrix: an easy-to-use collagen treatment for wound healing. AB - Collagen plays a major role in wound healing. Its presence is important in all stages of the healing process. Catrix is a new collagen wound-healing powder that has been shown to be effective in the treatment of wounds healing by secondary intent such as pressure ulcers, venous stasis ulcers and diabetic ulcers as well as second-degree burns and post-radiation dermatitis. Catrix has also been shown to be effective in the treatment of wounds unresponsive to conventional treatments. It promotes the growth of fibroblasts and keratinocytes in the wound, prevents loss of fluid from the wound and protects the wound from bacterial infections and other agents. Catrix is biodegradable and therefore does not require removal from the wound bed before re-application. PMID- 16245395 TI - Bird flu fears heading west. PMID- 16245396 TI - Anti-DNA idiotypes: from induction of disease to novel therapeutical approaches. PMID- 16245397 TI - Evaluating evidence for aging. PMID- 16245399 TI - [Therapeutic dilemmas in patients with a centrally-located pulmonary embolism confirmed by spiral CT-scan but with no cardiogenic shock]. PMID- 16245400 TI - The art of aging. Mental activity and creative pursuits may help stave off dementia and keep the mind clear. PMID- 16245401 TI - Hiking: not just a walk in the park. PMID- 16245402 TI - By the way, doctor. My systolic blood pressure is 40 points higher in winter than in the summer (160-180 versus 120-140 mm Hg). Do the seasons affect blood pressure. PMID- 16245403 TI - [Anencephaly findings in early abortion]. PMID- 16245404 TI - [Pseudobabinsky]. PMID- 16245406 TI - [Rei Medicae Praelectiones Theoricae Proemialium Quastionum. To expert mentor Pauli Manfredi 1695 Paulus Valle SBT: a collection of theoretical medicine of lessons of 1695 to the Rome Studium]. AB - In the Early Modern Age anatomy, physiology and clinical medicine become complimentary, subject to the same rational and systematic methodology. This contribution deals with a manuscript by Paulus De Valle, pupil of Paolo Manfredi, Lector of Medicina Theorica in the University of Rome. Manfredi's teaching follows modern authors: Harvey, on blood circulation and embryology, Bartholin on lymphatic vessels, Willis on anatomy and physiology of the nervous system. The manuscript sheds new light on the teaching of Medicine at Rome Studium in the 17th century. PMID- 16245405 TI - [Meningococcal meningitis without inflammatory reaction in the cerebrospinal fluid]. PMID- 16245408 TI - Tipranavir gets approved. PMID- 16245407 TI - Regulation of connexin expression after balloon injury: possible mechanisms for antiproliferative effect of statins. AB - Statins, the 3-hydroxy-3-methylglutaryl-cooenzyme A (HMG-CoA) reductase inhibitors, have been shown to inhibit the migration of macrophages and smooth muscle cell proliferation leading to an antiproliferative effect. Although this beneficial effect of statins has been suggested to be independent of lipid lowering properties, the possible mechanisms responsible for this action is largely unknown. Gap junctions, which serve as channels for direct intercellular exchange of ions, secondary messengers, and small signaling molecules, play an important role in tissue homeostasis and regulation of growth, differentiation, and development. This study was designed to test the hypothesis that expression of the component proteins of gap junctions, connexins 40 and 43 (Cx40 and Cx43), is upregulated in arteries subjected to balloon injury and that this upregulation can be suppressed by statin therapy. Male New Zealand white rabbits were subjected to injury in which an angioplasty catheter was introduced into the right iliac artery from the femoral artery under fluoroscopic guidance. Five groups of rabbits (n = 6 to 7) were treated for 2 weeks with one of the following: balloon injury (BL); BL+lovastatin (BL+L, 10 mg/kg/day); BL+fluvastatin (BL+F, 10 mg/kg/day); sham operation (Sham); and control (Con). Immunohistochemistry studies showed that Cx40 and Cx43 were expressed in normal smooth muscle cells (SMC) throughout the media. Reverse transcription-polymerase chain reaction and Western blot analysis showed that Cx40 and Cx43 mRNA and protein expression was elevated after injury (P < .001 for both proteins and both assays), and these elevations were suppressed by lovastatin and fluvastatin to a similar degree (P < .05 for both drugs and both assays). Immunostaining of Cx40 and Cx43 was consistently enhanced in the neointimal area after injury and lovastatin and fluvastatin reduced staining of these proteins in the lessened neointimal layer. Transmission electron microscopy revealed that there were abundant gap junctions between neointimal SMC as well as fewer and smaller gap junctions after statin treatment. Therefore, balloon injury causes upregulation of Cx40 and Cx43 in neointimal SMC. Lovastatin and fluvastatin suppress upregulated Cx40 and Cx43 expression and reduce neointimal proliferation, suggesting that Cx40 and Cx43 may play a role in statin-induced antiproliferative effect. PMID- 16245409 TI - Carotid plaque caused by usual risk factors. PMID- 16245410 TI - Same-sex marriage as HIV prevention tool. PMID- 16245411 TI - Carotid tonometry versus synthesized aorta pressure waves for the estimation of central systolic blood pressure and augmentation index. AB - OBJECTIVE: To assess the interchangeability of carotid tonometry and synthesized aorta pressure waveforms for estimating central systolic blood pressure (SBP) and augmentation index (AIx). METHODS: Tonometry waveforms were acquired with a custom built hardware and software platform in 276 subjects (179 men/97 women; aged 45.5 +/- 5.7 years; mean +/- standard deviation) at the radial (P(wf,ra)), brachial (P(wf,ba)), and carotid artery (P(wf,ca)). The P(wf,ba) was calibrated using systolic (SBP(ba)) and diastolic (DBP(ba)) sphygmomanometer pressure. The DBP(ba) and calculated mean (MAP(ba)) brachial pressure were subsequently used for calibration of P(wf,ra) and P(wf,ca). A central pressure waveform (P(wf,sao)) was synthesized from P(wf,ra) using a generalized pressure transfer function (TFF). The AIx and SBP were measured on P(wf,ra), P(wf,ca), and P(wf,sao). RESULTS: The SBP(ra), SBP(ca), and SBP(sao) were 138.5 +/- 16.8, 130.0 +/- 16.2, and 131.1 +/- 16.6 mm Hg, respectively. The SBP(ra) correlated well with the SBP(ca) (r = 0.93) and the SBP(sao) (r = 0.94), as did the SBP(ca) and the SBP(sao) (r = 0.97) with a mean bias of 1.35 +/- 3.90 mm Hg. The AIx derived from P(wf,ra), P(wf,ca), and P(wf,sao) were -20.8% +/- 14.5%, 12.4% +/- 13.9%, and 20.0% +/- 11.7%, respectively. The correlation between radial and carotid, and radial and central AIx was 0.72 and 0.94, respectively. The correlation between AIx derived from P(wf,ca) and P(wf,sao) was 0.75 with a bias of 11.0% +/- 14% (all correlations P < .001). CONCLUSIONS: The use of a generalized TFF in combination with well-calibrated radial pressure curves yields estimates of SBP in good agreement with carotid tonometry. Although AIx derived from a measured radial pressure curve correlates surprisingly closely with AIx measured on a synthesized aortic pressure curve, the correlation with a directly measured AIx on carotid signals is relatively poor. PMID- 16245412 TI - ADAP data available on web. PMID- 16245413 TI - Evidence for a combined approach to the management of hypertension and dyslipidemia. AB - The recognition that hypertension and dyslipidemia coexist more often than would be expected by chance and that their combination increases the risk of coronary heart disease (CHD) has important implications for patient management. Patients with cardiovascular disease (CVD) and healthy individuals with multiple CVD risk factors--including concomitant dyslipidemia and hypertension-are the primary focus of updated treatment guidelines for CVD prevention. There is a need for a treatment paradigm shift from the diagnosis and treatment of individual CVD risk factors to the assessment and management of total CVD risk. Goals for each CVD risk factor need to be intensified when multiple sources of risk are present. Patients who have no overt disease but who have multiple CVD risk factors are now understood to be at high risk and should be the target of aggressive primary preventive strategies to avert unnecessary morbidity and mortality. It is also becoming clear that CVD risk factors interact at the level of the endothelium to promote atherosclerosis. Furthermore, recent clinical trials, including those assessing atherosclerotic progression and CVD endpoints, have demonstrated the effectiveness of interventions targeted at multiple risk factors for CVD. PMID- 16245414 TI - [The Edward Smith papyrus]. PMID- 16245415 TI - The Elders' Oral Health Summit Proceedings. Boston, Massachusetts, 13-14 September 2004. PMID- 16245416 TI - [Place of ticlopidine in antiplatelet treatment]. AB - Ticlopidine is an inhibitor of platelets function in vivo. It inhibits platelets aggregation induced by ADP. Its mode of action has not been defined, but it appears that metabolites of ticlopidine are antagonists of the platelet ADP receptor. Inhibition of platelet aggregation is delayed until 24 to 48 hours after drug administration, achieving maximum after 3 to 5 days of treatment. Recovery of platelet function occurs from 4 to 14 days after discontinuation of ticlopidine. It is potent antiplatelet factor, possible to use (among other in view of low price) in secondary prevention of cardiovascular diseases as alternative drug, especially in prevention of thrombosis during coronary stent placement.. Use of ticlopidine is limited by numerous side effects forcing quite often to discontinuation of therapy. It seems, that better safety profile, and it probably larger efficiency shows different derivative of thienopiridyn- clopidogrel. PMID- 16245417 TI - [Renin-angiotensin-aldosterone system activity during head-up tilt testing in patients with vasovagal syncope]. AB - The stimulation of renin-angiotensin-aldosterone (RAA) system during tilt table test is caused by sympathetic nervous system activation by orthostatic stress and a serotonin release as well. In healthy individuals increase of plasma renin activity during test with maximal values on the peak of the test was described. OBJECTIVE: The aim of the study was to assess the activation of RAAS in patients with neurally mediated syncope during the tilt table test by means of plasma renin activity and serum aldosterone levels. MATERIAL AND METHODS: The study was carried out in 31 patients aged 39.4 +/- 15.0 years (18 women and 13 men) with neurally mediated syncope during tilt test. Plasma renin activity was assessed in the baseline conditions, immediately after the test and 10 minutes after the test using radioenzymatic assay. Aldosterone concentrations were measured radioimmunologically, twice: after 30 minutes supine rest and after the syncope. RESULTS: Plasma renin activity during supine rest was 2.2 +/- 2.4 ng/ml/h, rose after the syncope 2.5-fold to 5.2 +/- 4.5 ng/ml/h (p < 0.001 comparing to baseline) stayed on similar level approximately for the next 10 minutes--4.9 +/- 5.5 ng/ml/h (p = n.s.). In 11 patients (35%) 10 minutes after the test even further increase of PRA was observed. Serum aldosterone level increased significantly immediately after tilt test (90.0 +/- 72.9 vs 178.8 +/- 150.1 pg/ml, p < 0.01). CONCLUSION: Authors showed, that in patients with NMS plasma renin activity increases and this increase lasts for 10 minutes after the syncope and the concentration of aldosterone increases immediately after tilt test. PMID- 16245419 TI - [Doppler ultrasound assessment of blood flow parameters in hemodialysis fistulas]. AB - The number of patients requiring kidney replacement therapy is systematically increasing world wide. At the end of 2000 there were 8424 treated with dialysis in Poland. Ultrasound examinations allow assessment of dialysis fistula morphology and function. OBJECTIVE: Assessment of blood flow parameters in hemodialysis fistulas. Assessment of correlation between the analysed parameters. Optimization of number of parameters needed to characterise blood flow in dialysis fistulas. MATERIAL AND METHODS: Doppler ultrasound examinations were performed in 86 patients with hemodialysis fistulas. Various blood flow parameters were analysed. RESULTS: In analysed group a following values of analysed parameters were recorded: brachial artery cross sectional area was within the range 0.28-0.78 cm2; radial artery 0.05 - 0.39 cm2. The resistance and pulsatility index (RI and PI) of brachial artery were 0.32-1.00 and 0.39-3.56 respectively. Mean blood flow through brachial artery was 1082.3 ml/min and 946 ml/min through dialysis fistulas. CONCLUSIONS: Almost full correlation was found between volume of blood flowing through dialysis fistula and brachial artery, brachial and radial arteries and between PI and RI. The volume of blood flowing through brachial artery depends mostly on brachial artery cross section area and mean blood flow velocity. In patients with dialysis fistulas located on the arm or forearm blood flow measurements may be performed within the brachial artery. PMID- 16245418 TI - [The effect of fenspiride on the number of exacerbations and the time of first exacerbation in patients with chronic bronchitis]. AB - The aim of the work was evaluation of efficacy of fenspiride b.i.d. on the number of exacerbations and the time to the first exacerbation in patients with chronic bronchitis. MATERIAL AND METHODS: Randomized, multicentre study controlled versus placebo was carried out in 12 centers in Poland. All patients, 89 females and 68 males aged between 20 and 74, were treated with fenspiride at the dose of 160 mg/day for a period of 6 months. The following symptoms were recorded every month in order to evaluate the therapeutic efficacy: sputum quality and quantity, cough intensity, dyspnea and bronchospasm. Based on these symptoms diagnosis of exacerbation was performed according to American Thoracic Society criteria. RESULTS: Quality and quantity of sputum and cough significantly improved in the fenspiride group (comparing to the placebo group p= 0.027 and p = 0.049 adequately for sputum and cough). A significant difference between groups was observed in the number of exacerbation episodes and their duration. In the fenspiride group there were 0.53 episodes of exacerbation compared with 1.12 episodes in the placebo group (p = 0.038). Mean duration of exacerbation was 3.3 days in the fenspiride group and 7.3 days in the placebo treated patients (p = 0.034). Time to the first exacerbation differed between groups, but this difference was not statistically significant. Number of side effects observed did not differ between groups. CONCLUSION: Fenspiride treatment was assessed as relatively effective in terms of influence on exacerbations, and well tolerated during six month therapy. PMID- 16245420 TI - [The effect of low magnetic field on select parameters of blood coagulation]. AB - Low frequency magnetic field causes the biological effects in organisms--in individual systems and organs. OBJECTIVE: The purpose of our study is to analyse the influence of the low magnetic field used in magnetotherapy on prothrombin time, factor Xa activity and level of platelets in experimental animals. MATERIAL AND METHODS: The examination was carried out on rats which were subjected to the activity of low magnetic field. The examination of prothrombin time in Quick's method (the international normalized ratio--INR was counted), activity of factor Xa by spectrophotometric method (lambda = 405 nm) were performed. The level of platelets before and after 14, 28 days of the exposition of magnetic field and after 21 days of exposure to magnetic field was assessed. RESULTS: The extension of prothrombin time (a decrease of INR), decrease activity of factor Xa and decrease platelets level was observed in experimental animals after the exposure to low magnetic field (p < 0.05 for all examined parameters). CONCLUSION: Low magnetic field used in magnetotherapy causes the changes in blood coagulation of experimental animals. PMID- 16245421 TI - [Serum and cerebrospinal concentrations of sICAM-1 sICAM-2, sICAM-3 in neuroborrellosis and tick borne encephalitis--preliminary report]. AB - OBJECTIVE: The aim was to evaluate the concentrations of the soluble forms of ICAM-1, ICAM-2, ICAM-3 in the serum and cerebrospinal fluid (CSF) of patients with neuroborreliosis and thick borne encephalitis before and after therapy in comparison with the control group. MATERIAL AND METHODS: We examined 30 patients, 10 in each group: neuroborreliosis-group I (NB), tick borne encephalitis--group II (TBE) and in the control group (group K). The diagnosis of neuroborreliosis and TBE based on the clinical features was confirmed by ELISA assays: FSME Virus/TBE Virus test (VIRION, Germany) for TBE and Borrelia IgM and IgG Recombinant (Biomedica Austria) for NB. The assays of sICAM-1, sICAM-2, sICAM-3 (ELISA, Bender Med System, USA) were performed twice in group I and II: before and after 3-4 weeks long treatment and once in control group. RESULTS AND CONCLUSIONS: Increased concentration of soluble forms of ICAM-1, ICAM-2, ICAM-3 in CSF suggest their important role in inflammatory process of viral and bacterial origin. In NB group, the serum concentrations of sICAM-1, sICAM-2 were significantly increased before and after treatment in comparison with control as well as with the analogous test results in TBE group. It may suggest NB as the part of systemic inflammation. The CSF concentration of sICAM-2 decreases after treatment in NB group in comparison with the analogous test results of TBE group. The increased CSF concentration of sICAM-2 in TBE group when compared to the CSF concentration in NB group suggest slow recovery and still persisting immunological activation in this group, even when the neurological symptoms disappeared. Increased concentrations in CSF in both diseases indicate intrameningeal activity of lymphocytes and may be a useful marker of inflammation. PMID- 16245422 TI - [Prognostic role of troponin I level in ischemic stroke--preliminary report]. AB - THE OBJECTIVE: The purpose of this research was to establish if determined at the reception level of troponin I could be such kind of factor. Presented data represent preliminary report of this study. MATERIAL AND METHODS: The analyzed data base on 196 patients with IS: 166 discharged from hospital and 30 with clinical course terminated by death. The IS was confirmed by computed tomography (CT). The conducted recording consists of data like e.g.: age, sex and risk stroke factors. Also the following factors were evaluated on admission: state of consciousness, level of neurological deficiency according to Scandinavian Stroke Scale (SSS). After half of year again SSS level was evaluated. RESULTS: The studied groups differed in level of neurological deficiency determined on admission. The direct mortality within IS group and increased level of troponin I represents 20.90% and with correct level of troponin I represents 12.40% (p=NS). The half year mortality group recording has not been considered yet because research is still continuing and database is continually updated. CONCLUSION: The increased level of troponin I on admission is not a prognostic factor of direct mortality within IS patients. We noticed that occurrence of disturbances in patient's EKG places those patients in the group of increased risk of death. PMID- 16245423 TI - [Correlations between clinical course and investigation results of central conduction time of somatosensory potentials evoked from tibial nerves in patients with cervical myelopathy treated by surgery]. AB - OBJECTIVE: The aim of study was to determine if abnormal somatosensory evoked potentials (SSEPs), specifically central conduction time (CCT) correlated with the degree of cervical myelopathy surgically treated or with clinical improvement following surgery. MATERIAL AND METHODS: Thirty patients (women, n=12 and men, n=18) with cervical myelopathy with neurological signs of cervical pyramidal tracts lesion and gait difficulties treated in 7-th Navy Hospital were studied. In our group in MR investigation we observed 26% discopathy, 56% cervical spondylosis and 43% cervical discopathy and spondylosis. Patients were treated surgically in cervical region. SSEP from tibial nerves data especially N22 to P40 interlatency (CCT) and basic everyday motional ability (by Dudek and Michno) were collected pre-operatively and 6-month after surgical treatment. RESULTS: Preoperatively, 43% (n=13) patients had normal values of CCT No patients had worsened CCT value after surgery. Six months after surgical treatment we observed abnormal in 13% (n=4) and normal values of CCT in 86% (n=26). In the second group we observed summary 60% (n=18) decrease of basic everyday motional ability. CONCLUSION: In 56% patients with cervical myelopathy abnormal SEP's from tibial nerve's abnormal CCT we observed. There is significant reduction of abnormalities SEP's from tibial nerve's after surgery. Abnormal SSEPs before the surgery correlates with the severity of myelopathy and improvement in SSEPs following surgery correlates strongly with clinical improvement. PMID- 16245424 TI - [Depression and anxiety in patients with adrenal tumor incidentally found on abdominal imaging performed for other indications]. AB - OBJECTIVE: The aim of this study was to assess the level of anxiety and depression in patients with non-secreting adrenal incidentaloma. MATERIAL AND METHODS: The study group comprised 26 patients (16 women, 10 men, mean age 45, SD 11 yrs). The Polish version of the State-Trait Anxiety Inventory was used to assess anxiety level, and the Beck Depression Inventory was used to measure depression symptoms. RESULTS: The anxiety level was increased (median: 45.6 for state, and 46.5 for trait), and mild depression was present, with 13.5 scores on Beck's scale. CONCLUSIONS: Adrenal incidentaloma is associated with increased anxiety and depression. PMID- 16245425 TI - [Evaluation of resistance of Pseudomonas aeruginosa isolates to select antibiotics]. AB - P. aeruginosa are opportunistic microorganisms which caused serious hospital infections. The resistance of 763rd strains of Pseudomonas aeruginosa to selected antibiotics (beta-lactams, aminoglycosides, quinolones) were tested by disk diffusion method. The least of Pseudomonas aeruginosa strains were resistant to imipenem and ceftazidime. Multiresistant strains of Pseudomonas aeruginosa are often isolated from clinical specimens. PMID- 16245426 TI - [The secretory activity of blood platelets--beta-thromboglobulin and platelet factor 4 in echinococcosis]. AB - Parasitic infection can cause local or systemic inflammatory process. Antibody dependent cellular cytotoxicity, with eosinophils as the effector cells, is the major mechanism of antiparasitic defence. The cytotoxic activity against antibody coated parasites is also demonstrated by blood platelets. Platelets initiate and maintain inflammatory processes, become activated due to contact with immunocompetent cells or due to a direct contact with a parasite. THE OBJECTIVE: The aim of the examination was to evaluate the cytotoxic activity of platelets in patients infected with Echinococcus granulosus. MATERIAL AND METHODS: The study was conducted on a group of 24 patients (aged 26-69) infected with Echinococcus granulosus, 19 women and 5 men. The patients were treated with albendazole (Zentel) and did not give consent to surgical procedure. Blood for analysis was collected prior to treatment. Patients infected with E. granulosus were subjected to the examination of platelet activation through the assessment of beta thromboglobulin and platelet factor 4 concentrations. The concentration of beta thromboglobulin and platelet factor 4 were assayed using a set of ASSERACHROM (Boehringer Mannheim) according to the immunoenzymatic method with labelled antibodies. Platelet count and their morphological parameters were determined with haematologic analyser Technicon H-3 System. Although platelets do not get into a direct contact with the parasite, an increase is observed in the concentrations of beta-thromboglobulin (16.09 +/- 2.68 IU/ml) and platelet factor 4 (8.47 +/- 3.32 IU/ml), which may indicate the involvement of platelets in the parasitic disease. PMID- 16245427 TI - [Limulus amebocyte lysate test use in clinical diagnostic in children with infectious diseases--preliminary report]. AB - OBJECTIVE: The aim of this preliminary study was (1) to evaluate the usefulness of the Limulus amebocyte lysate (LAL) test as a diagnostic tool in pediatric clinic, and (2) to examine the serum ET levels in hospitalized patients. MATERIAL AND METHODS: Bacterial blood endotoxin (ET) was measured in 50 children with infections of different etiology and localization. A Limulus amebocyte lysate (LAL) test was used. CONCLUSIONS: The usefulness of LAL test in routine diagnostic procedures in hospitalized children with infections was not proven so far. However, the LAL test appeared useful in the urinary tract infection (UTI) cases in which significant elevation of serum ET levels were demonstrated. Moreover, it was confirmed that LAL test can be performed as well in serum: the mean values of serum ET levels in our patients were comparable with published ET plasma values in adults. PMID- 16245428 TI - [Acceptance of screening colonoscopy in the prevention of colorectal cancer in relation to some demographic factors]. AB - Colorectal cancer is a quite common neoplasm. Screening colonoscopies and polypectomies can decrease the incidence of that neoplasm by 75%. Discovering the most frequent reasons and most convincing sources of information about the examinations is important for success of screening programme. OBJECTIVES: The aim of this study was to find the connection between some demographic factors, the source of information about the examination and the consent for colonoscopy. MATERIAL AND METHODS: Two groups of subjects, aged 40-65, were included: 1000 subjects, who gave their consent for colonoscopy and 200 subjects, who refused the examination. The sources of information were two kinds of questionnaires: fulfilled before colonoscopy and in case of lack of consent for the examination, which included demographic data, the reason of undergoing or refusal of colonoscopy and the source of information about the examination. RESULTS: The higher educated were the subjects, the more often they agreed for the examination; among subjects after university 87%, and among subjects with elementary education 71% gave their consent for colonoscopy. The most convincing sources of information were the medial sources; 100% of subjects, who found out about the examination from television and radio, gave their consent for colonoscopy. The most frequent reason of undergoing the examination was prophylaxis (54.6%), and the most frequent reasons of refusal was fear of pain (33.5%). CONCLUSIONS: Educational level and the source of information about the examination are the factors that have an important influence on undergoing screening colonoscopy. Identifying the influence of some demographic factors on the consent for colonoscopy and the efficacy of each source of information seems to be very important for the success of screening colonoscopy programme and therefore for discovering adenomatous polyps and colorectal cancer. PMID- 16245429 TI - [The efficacy and safety of trazodone in the treatment of adjustment disorders- naturalistic research]. AB - There are a great number of traumatic and stressful events that people are exposed to in the contemporary world. The response to them is the subjectively felt stress and numerous emotional reactions with anxiety and lowering of the mood. These symptoms substantially limit the ability of social and vocational performance. Their appearance allow to diagnose adjustment disorders. AIM OF THE STUDY: The evaluation of therapeutic efficiency and safety of using trazodone in treatment of adjustment disorders with the particular focus on dominant symptoms in this illness that is anxiety, lowering of the mood, low self-esteem, sleep disorders and also aggression level. MATERIAL AND METHODS: 52 people of both sexes participated in this study. They were diagnosed with adjustment disorders and underwent some treatment in psychiatric surgery. The completed Hamilton's Depressive Scale and Anxiety Symptoms Scale COVI. Clinical condition of the patients was examined with Clinical General Impression Scale. The level of self Acceptance was measured with ACL test. The examination lasted 56 days. RESULTS: Post hoc statistical tests allowed to form the idea that in the case of depression the intensity of the symptoms decreased significantly on the 14th day of the therapy. However, the anxiety symptoms decreased dramatically on the 7th day. CONCLUSION: (1). Trazodone turned out to be an efficacious medicine in the therapy of adjustment disorders. On the 56th day of the study the illness of 65.4% of patients was in remission, 26.9% of them responded to treatment, and 7.7% of them didn't. (2). Trazodone on the 7th day of therapy diminishes considerably the anxiety intensity and sleep disorder, and on the 14th day of the therapy it has an antidepressive effect. (3). As result of treating patients with trazodone the level of self-acceptance measured by ACL test has increased. PMID- 16245430 TI - [Off-pump coronary artery bypass grafting and left ventricular aneurysm resection -a case report]. AB - We present the case of a 55-year-old patient with coronary heart disease of class III of the Canadian Cardiovascular Society (CCS), after cardiac antero-lateral wall infarction at unknown time. Coronarography revealed 90% occlusion of three branches of left coronary artery. Ventriculography showed dyskinetic aneurysm of apex and anterior wall without presence of thrombus. On the basis of the performed examination the patient was qualified to off-pump coronary artery bypass (OPCAB) and left ventricular aneurysm resection (LVAn). On the beating heart with the use of Octopus II stabilizer distal (side to side) anastomosis was performed to marginal branch and (end to side) to circumflex as a jump graft. Subsequently left internal thoracic artery to left anterior descending branch anastomosis was performed. And then, on the beating heart on the two Teflon pads entry of aneurysm was closed by the mattress sutures. Next the aneurysm was resected and closed with additional continuous suture. There were non operative and post-operative complications. PMID- 16245431 TI - [Abdominal migraine as a cause of chronic recurrent abdominal pain in a 9-years old girl--case report]. AB - Abdominal migraine is a rarely recognized functional intestinal disorder, manifesting as recurrent paroxysmal abdominal pain of neurogenic origin. The authors describe the 9-years old girl referred to the hospital because of chronic paroxysmal abdominal pain. She did not improve after medication used commonly in functional abdominal disorders (drotaverine, mebeverine, trimebutine). On the ground of various investigations organic causes of abdominal pain were excluded. Carefully completed anamnesis, as well as precise description of the clinical picture of abdominal pain attacks, has lead to the diagnosis of abdominal migraine. According to advice of neurologist the treatment with amitriptyline was introduced. Thereafter a significant improvement was observed. Abdominal migraine has to be taken in to account when diagnosing chronic abdominal pain in children. PMID- 16245432 TI - [Adrenal tuberculosis as a cause of primary adrenal insufficiency--case report]. AB - Adrenal tuberculosis is relatively infrequent cause of primary adrenocortical insufficiency in developed countries. Adrenal involvement is most often the result of hematogenous spread of the pulmonary tuberculosis. Isolated adrenal tuberculosis, especially with enlargement of adrenal glands can cause diagnostic problems and requires differentiation from primary or secondary neoplastic disease. In this paper we present a case of 61-year-old man with several months history of adrenocortical insufficiency without signs of pulmonary tuberculosis. Computed tomography scan revealed asymmetrical mass-like enlargement in adrenal glands. Despite of consecutive investigations, the diagnosis remained uncertain. Because of the possibility of neoplastic process of unknown origin, the patient was qualified for surgical exploration during which both enlarged glands were removed. The diagnosis of tuberculosis was made on microscopic examination. PMID- 16245433 TI - [Successful pharmacological treatment of hyperinsulinemic hypoglycemia with verapamil and amlodipine--case report]. AB - We present a case of an 80-years-old female patient admitted to hospital because of recurrent losses of consciousness due to hypoglycemia, and because of hypertension. In view of the clinical picture (Whipple's triad) a suspicion of hypoglycemia due to an insulinoma of the pancreas aroused. The suspicion was confirmed by a positive fast test. The test was terminated after 22 hours with a glucose concentration of 2.0 mmol/l (36 mg/dl) and an insulin level of 16.19 microU/ml. No visual diagnostic was performed due to the lack of the patient's consent, thus no diagnosis of a tumor of the pancreas could be established. Hypertension was controlled with two calcium channel inhibitors: verapamil and amlodipine were administered orally, in doses of 120 mg and 10 mg per day, respectively. Both medications were chosen because of intolerance to other anti hypertensive agents. During two-weeks treatment an increase of glucose concentrations and no hypoglycemic symptoms were observed. The patient did not agree to surgical intervention and therefore she was discharged in good general condition. CONCLUSION: we observe the efficacy of verapamil and amlodipine administered simultaneously in the medical treatment of endogenous hyperinsulinemic hypoglycemia. Their efficacy in the medical treatment of insulinoma requires further research. PMID- 16245434 TI - [Surgical restoration of left ventricle after myocardial infarction]. AB - Heart failure is currently one of the biggest problems of all heath care systems in the world and will become even more important. In about 50% of the patients, heart failure is a consequence of myocardial infarction. The surgical reconstruction of the left ventricle after anterior myocardial infarction became the first effective method of treatment which can be applied in many patients. After anterior myocardial infarction the apical part of the ventricle dilates, becomes more spherical with thinned wall. Dilated, spherical apex results in rise of wall tension of the transition zone between the apex and the proximal part of the left ventricle. This increases oxygen demand and reduces coronary blood flow. This phenomenon is responsible for severe angina pectoris and explains why myocardial infarction is the most common cause of death of patients with LV aneurysm. Increased wall tension of the transition zone leads to fibrosis, further dilatation and decrease of contractility of the left ventricle. Only surgical reconstruction of the left ventricle can stop this progressive process. The aim of surgery is to decrease the wall tension by restoring the normal elliptical shape, geometry and volume of the left ventricle. Results of this operation are very promising. Perioperative mortality is about 5% and 5-year survival 80%. PMID- 16245435 TI - [The Churg-Strauss syndrome and antileukotriene agents]. AB - The Churg-Strauss syndrome (CSS), characterized by asthma, eosinophilia and systemic vasculitis, was initially described in 1951. Criteria of recognition were defined by: Churg and Strauss (pathologic criteria), Lanham and coworkers (clinical criteria) and American College of Rheumatology (differentiation from others forms of vasculitis). CSS is often a triphasic illness and main symptoms derive from an affection of respiratory, cardiovascular, nervous and digestive systems. The pathogenesis of CSS is not clear, but it is suspected that antileukotriene agents, used in the treatment of asthma, are probably involved in. However, this association has not been unequivocally proved until now. PMID- 16245436 TI - [Predictors of successful renal artery revascularization in atherosclerotic renovascular disease]. AB - Atherosclerotic renovascular disease (ARVD) is defined as a reduction in glomerular filtration rate in patient with significant renovascular bilateral occlusive disease or unilateral in a solitary kidney. ARVD is a frequent and potentially avoidable cause of end stage renal failure and the need for replacement therapy among person above 50 years old. Use of balloon angioplasty with stenting or surgical repair has been shown to improve renal potency, but there is no clear evidence that it prevents further progressive decline of renal function or blood pressure, compared to medical therapy alone. It is now recognized that severity of histopathologic damage is an important determinant and predictor of renal functional outcome. Proteinuria increases with declining renal failure and reflects the severity of parenchymal damage. Proteinuria and high renal resistance index are associated with glomerular damage and altered intrarenal perfusion. They aren't linked to renal artery stenosis (RAS) grade. Recent study indicate that abrupt decline in renal function reflects reversibility after revascularization. This should probably be undertaken in RAS patients with rapidly deteriorating renal function, refractory hypertension and in whom plasma creatinine concentration has increased by >20% during one month long administration of angiotensin-converting enzyme inhibitor. Renal biopsy may be useful to evaluate patient who will have advantage from revascularization. PMID- 16245437 TI - [Glomerulonephritis in neoplastic disease]. AB - Glomerulonephritis are known to be associated with cancer disease. Although these association is well documented but the number of conditions must be fulfilled to establish a causal relationship between the malignancy and glomerulopathy. Secondary membranous glomerulonephritis is relatively often noted type of glomerulopathy, but other types have been mentioned, especially IgA nephropathy, minimal-change disease, rapidly progressive glomerulonephritis and anti-GBM disease in the case of neoplastic obstruction of urinary tract. The most frequent types of malignancies are pulmonary, renal and gastrointestinal solid tumours. The pathogenesis is immunological due to the involvement of tumor associated antigens, re-expressed fetal antigens, and/or viral antigens. Immune complexes (circulating or formed in situ) containing tumor-associated antigens are suspected of causing glomerular injury. The diagnosis of malignancy may be established either before, concomitant or after the development of nephrotic syndrome. Slow remission of nephrotic syndrome is sometimes observed after resection or successful treatment of the tumor, and relapse has been reported during the relapse of cancer. The non-specific treatment of nephrotic syndrome is recommended and the immunosuppression generally should be avoided because this may exacerbate the malignancy. Sometimes also antiviral therapy can be considered. Patients with nephrotic syndrome due to membranous glomerulonephritis, aged over 50, should be carefully screened for malignancy. The tumor prognosis determines patient outcome. PMID- 16245438 TI - [Heat shock proteins and their characteristics]. AB - The main adaptable response to increased temperature is heat shock response resulting in induction of proteins called heat shock proteins (HSP). They are present in all cells under proper growth conditions and they create 5-10% of the whole protein contents. HSP were divided into five basic groups according to their approximate molecular mass, expressed in kDa and called respectively: HSP 100, HSP 90, HSP 70, HSP 60 and small HSP. Heat shock proteins can act like antigens in many infectious diseases. Immunological response against proteins from HSP 60, HSP 70 and HSP 90 families was observed in diseases caused by bacterial and protozoan pathogens. It is known that ischemia and reperfusion activate HSP genes transcription in heart cells of various experimental animals. Human and Chlamydia pneumoniae HSP 60 were found in patients with stable coronary disease. Hence many researchers connect the increase of ischaemia with the passed infection caused by Chlamydia pneumoniae, which can influence the origin or development of atheromatous plaque in the vascular wall. HSPs play an important role in hyperthermic therapy commonly used together with irradiation. Moreover, works on the possibility of HSP application to delay of disease process in neurodegenerative diseases, such as Parkinson or Alzheimer diseases are conducted. The paper presents characteristics of heat shock proteins, role in the state of health and disease and possibilities of their usage in monitoring and/or treatment of diseases, e.g. cancers. PMID- 16245440 TI - [The healthcare worker as a source of hepatitis C virus infection]. AB - The paper discusses risks associated with the possibility of transmission of hepatitis C virus (HCV) infection from an infected healthcare worker to a patient. Reports describing infections where a healthcare worker was the source of HCV infection to patients are summarized and guidelines concerning approach to infected healthcare workers in other countries are presented. Surgeons performing exposure-prone procedures are almost the only source of infection to patients, provided that universal precautions are respected. Exposure prone procedures occur most commonly during gynaecologic, orthopedic, thoracic operations. At present routine testing of all healthcare workers for HCV infection is not justified. Some experts recommend screening for HCV infection surgeons performing exposure-prone-procedures. Although the present risk of HCV transmission from a HCV infected (HCV-RNA positive) healthcare worker to a patient is small, estimated to be about 1/7000 surgical procedures, some countries recommend that healthcare workers infected with HCV should be restricted from undertaking exposure prone procedures. PMID- 16245439 TI - [The course of hepatitis C infection after liver transplantation]. AB - HCV infection is an important health problem all over the world. Hepatitis C is the main reason for liver cirrhosis and liver failure which is one of most common indication for liver transplantation (OLT). HCV-associated liver cirrhosis appears at 20-25% of infected patients within 20 years. Recurrence of HVC infection nearly universally follows transplantation and may lead to graft loss. Acute hepatitis appeared in 75% of patients. The rate of HCV cirrhosis on the graft is variable from 8 to 25% at 5 years. Antiviral treatment with combination therapy interferon-ribavirin gave promising results. Attempts to introduce prophylactic post-transplant antiviral treatment are still under evaluation. This paper presents clinical and therapeutical issues of recurrent hepatitis C after liver transplantation. PMID- 16245441 TI - [Serum lipids concentration and colorectal cancer]. AB - The authors present current concepts concerning the relationships between plasma lipid concentration and incidence of colorectal cancer. Pharmacological reduction of cholesterolaemia may decrease mortality of cardiovascular causes' simultaneously increasing mortality of non-vascular causes. Discussion concerns the idea of preclinical action of the neoplasm on lowering cholesterol level as well as hypothesis of increased biliary secretion of cholesterol metabolites, that may have cancerogenic effect. Cholesterol plasma concentration and its impact on aetiopathogenesis of colorectal cancer still need to be investigated. PMID- 16245443 TI - [Diagnostic value of total and lipid-bound sialic acid in malignancies]. AB - Malignancies are the second cause of death among people after cardiovascular diseases. The early detection of tumors could increase the possibility of a favorable treatment outcome. Many biochemical tumor markers have become known during the last forty years, but each of them has some clinical limitations. Recently, the main attention of researchers is focused on tumor-derived compounds as possible markers of neoplasia. In this paper, the diagnostic value of serum total sialic acid (TSA) and lipid-bound sialic acid (LSA) has been summarized in malignant disease including the diagnosis, staging, prognosis and follow up of cancers. The clinical studies showed that TSA and LSA concentrations in serum were significantly elevated in different types of cancers compared to values of healthy people and with nonmalignant diseases. In the initial phase of malignant diseases, TSA and LSA as single tests for detection of cancers are not useful according to low diagnostic sensitivity. TSA and LSA concentrations in serum allow with some probability to assay the staging of cancer. LSA is more sensitive and specific test than TSA. Besides, serum TSA and LSA levels good reflect effectiveness of therapy. The decrease of concentration indicates on tumor regression (effective therapy) and the increase means the progression of disease (failing therapy). TSA and LSA can be useful biochemical indicators for staging, prognosis, monitoring of effectiveness treatment of cancers and early detection of recurrence or metastases, especially in combination with other tumor markers. PMID- 16245442 TI - [Butcher's Broom, in the treatment of venous insufficiency]. AB - Numerous extracts of the plants, natural compounds and their derivatives, acting on the venous system, including horse-chestnut seed extracts aescin, rutin, troxerutin, diosmin and hesperidine. They have a long tradition in herbal medicine for their venotonic and anti-oedematous properties. This review is concerning Rusci aculeati rhizoma, which recently taken in Poland on the symptoms of chronic venous insufficiency and hemorrhoids. PMID- 16245444 TI - [Hypoxia in malignant tumors the target for clostridial therapeutic gene strategies]. AB - In the fast proliferating malignant tumors which are characterized by abnormal vascularisation develop non oxidized regions of a low oxygen partial pressure (pO2). Because these tumors are unsusceptible to a conventional therapy they were chosen for clostridial therapeutic gene strategies. There were presented different strategies using clostridia as selective vectors of destructive genes, e.g. cytosine deaminase and nitroreductase activating in reduced microenvironment of tumors--nonactive prodrugs towards strong cytostatics, destroying hypoxic tumor cells. In general, they form a basis for CDEPT strategy (clostridial directed enzyme prodrug therapy) and moreover in connection with antivessel cytostatic they form COBALT strategy (combination bacteriolytic therapy). In another strategy genetically modified clostridia are transferred to the gene of TNF alpha cytokine which production regulates fractionated ionizing radiation activating a promoter include in the clostridial vector. Sat last, there were discussed new problems connected with clostridial therapeutic gene strategies. PMID- 16245445 TI - [The role of sentinel node biopsy in breast cancer surgery]. AB - Mastectomy with axillary lymph node dissection remains the routine surgical treatment of breast cancer in Poland. Lymph edema of the upper extremity is one of the major long-term complications of axillary dissection. Axillary lymph node status is the most valuable prognostic indicator and decision factor on adjuvant chemotherapy or radiotherapy for breast cancer patients. Level I and II axillary lymph node dissection provides prognostic information, maintains local control in the axilla and determines the need for adjuvant systemic treatment, but it is also associated with 30% rate of lymph edema. Multiple studies confirm that sentinel lymphadenectomy accurately stages cancer advancement and is associated with less morbidity than axillary dissection. Over 40% of breast cancer patients in Japan are submitted to breast conserving therapy with sentinel node biopsy. In our opinion sentinel node biopsy may be accepted as an alternative staging procedure for the axilla in breast cancer. Sentinel node biopsy is especially valuable tool for breast cancer patients undergoing breast sparing surgery (IIA), due to excellent cosmetic outcome, minimal morbidity and high degree of histological accuracy associated with the procedure. This article reviews current literature in breast conserving therapy and sentinel node biopsy. Author would like to thank to Professor Kenji Ogawa, Chairman of Surgical Department of Tokyo Women's Medical University Daini Hospital, Professor Fujio Kasumi Chief of Breast Surgical Department of Cancer Institute Hospital and the Japan Society for the Promotion of Science for the scientific support during research visit in Tokyo. PMID- 16245446 TI - [The biomedical aspects of the non-antioxidative action of vitamin E and its resulting role in therapy]. AB - The review of current informations connected with the non-antioxidative applications of vitamin E in therapy was performed. After explanation indispensable structural and nomenclatural questions, were have talked over was for above mentioned uses: biological activity of this vitamin, food sources and the metabolism. It was presented the systemic non-antioxidative functions and the resulting actions from here with range of prevention. The attention on day requirement, the states of deficiency and hypovitaminosis, and also on normal concentration in blood plasma was emphasized. PMID- 16245447 TI - [Do you have the "grip" in your grip?]. PMID- 16245448 TI - [Venereal syphilis. "Return" of an old infectious disease]. PMID- 16245449 TI - [Tips and tricks, what is not in the package insert. Hints for customers for proper handling of various dosage forms]. PMID- 16245450 TI - [Type 2 diabetes with alcohol abuse]. PMID- 16245452 TI - [Intradermal administration of a strong vaccine]. PMID- 16245451 TI - [Should children be given aromatic oils?]. PMID- 16245453 TI - [Neuraminidase inhibitors for prevention of influenza?]. PMID- 16245454 TI - [Oseltamivir resistant influenza in children]. PMID- 16245455 TI - [Are high doses of vitamin E harmful?]. PMID- 16245456 TI - [The effects of thiazolidinedione from the cardiologic viewpoint]. PMID- 16245457 TI - [Better metabolic control with patient-focused insulin administration]. PMID- 16245458 TI - [Medicinal coal shortens the duration of diarrhea]. PMID- 16245459 TI - [Possibilities for treatment of scars]. PMID- 16245460 TI - [Silver coated urinary catheters. Clinical effectiveness and cost-benefit analysis]. PMID- 16245461 TI - [Burns. Cold water immersion]. PMID- 16245462 TI - [Dementia of trisomy 21. Improvement with donepezil]. PMID- 16245463 TI - [Multiple sclerosis. A new oral immunomodulator]. PMID- 16245464 TI - Calcium hydroxide: a review. AB - Calcium hydroxide is a multipurpose agent, and there have been an increasing number of indications for its use. Some of its indications include direct and indirect pulp capping, apexogenesis, apexification, treatment of; root resorption, iatrogenic root perforations, root fractures, replanted teeth and interappointment intracanal dressing. The purpose of this paper is to review the properties and various indications for the use of calcium hydroxide. PMID- 16245465 TI - The use of Traditional Chinese Medicine in managing oral health--Hong Kong: one country, two systems. AB - OBJECTIVES: To determine knowledge, use, and interest in traditional methods to manage oral health among Hong Kong adults. DESIGN: A cross sectional epidemiological survey. PARTICIPANTS: A random sample of 500 Hong Kong adults. METHODS: Telephone interviews about Traditional Chinese Medicine (TCM) approaches to mange oral health, employing a battery of 50 questions over a one month period in 2002. RESULTS: Most completed the interview (96%, 481/500). Over half (52%, 250/481) had some knowledge about using TCM to manage oral health. Approximately a third (32%, 155/481) reported using TCM to manage their oral health in the past, most frequently for oral mucosal lesions (65%, 101/155) or for periodontal reasons (62%, 96/155). TCM was used predominately in combination with western dentistry or as a last resort (55%, 86/155). Most (89%, 428/481) expressed an interest in TCM approaches to managing oral health, particularly if administered by their dentist (54%, 259/481), and expressed many reasons for such interest. Socio-demographic variations in knowledge, use and interest in the use of TCM to manage oral health were apparent (p<0.05). CONCLUSION: Knowledge, use and interest in traditional based approaches to managing oral health is widespread in Hong Kong, and is associated with age and educational attainment. PMID- 16245466 TI - The shortened dental arch--an assessment of patients' dental health state utility values. AB - OBJECTIVES: The aim of this study was to determine how patients value the potential outcomes from different treatments for the shortened dental arch (SDA). METHODS: 110 partially dentate patients were interviewed and presented with illustrative models as well as a description of the process and likely outcome of six possible treatments for the shortened dental arch. The patients were asked to indicate on a standardised visual analogue scale how they would value the health of their mouth if they had received each of the treatments described. RESULTS: With a utility value of 0.0 representing the worst possible health state for a mouth and 1.0 representing the best, the mean utility value for 'no treatment' for a SDA was 0.28 (sd=0.29). For a cobalt-chromium based removable partial denture, an acrylic-resin based removable partial denture, implant treatment, a resin-bonded cantilevered bridge and a conventional cantilevered fixed bridge the utility values were 0.42 (sd=0.3), 0.49 (sd=0.31), 0.53 (sd=0.35), 0.63 (sd=0.26) and 0.64 (sd=0.28) respectively. CONCLUSIONS: The treatment outcome from a cantilevered bridge attracted the highest mean utility value. Participants did not value implant treatment as highly as fixed bridgework, but they did rate the utility of implants more highly than the utility value of removable dentures. However, the patients placed a very low value on the outcome from 'no' treatment. Thus, the appropriateness of the shortened dental arch as an oral health goal can be questioned. PMID- 16245467 TI - A pilot study of treatment of periodontal-endodontic lesions. AB - OBJECTIVE: A pilot study to evaluate a standardised treatment protocol for combined periodontal-endodontic lesions (involving staged endodontic and periodontal treatment). PARTICIPANTS: Nine patients with a diagnosis of a combined periodontal-endodontic lesion. METHODS: At baseline, root canal treatment (RCT) was performed by a standard protocol, and simple scaling/oral hygiene instruction provided. After one month, a standardised episode of non surgical periodontal treatment was undertaken to address residual pocketing. Clinical measurements, including probing depths, attachment levels and bleeding on probing were recorded at baseline, month one and month three. Long-cone periapical radiographs taken using standardised projection geometry at baseline and month three were analysed for bone changes using digital subtraction radiography (DSR). RESULTS: During the study, one patient's affected tooth was extracted. From baseline to month three, there were statistically significant mean probing depth reductions (delta= 0.95mm, 95% CI= 0.20mm, 1.70mm; p= 0.02), mean attachment gains (delta= 1.13mm, 95% CI= 0.29mm, 1.96mm; p= 0.02) and reduction in mean bleeding on probing (delta= 29%, 95% CI= 10%, 49%; p= 0.01). DSR analysis revealed that between month 0 and month three, four teeth demonstrated bone gain, two teeth exhibited bone loss and two teeth showed no change. The mean bone change was in favour of bone gain but failed to achieve statistical significance (p>0.05). CONCLUSIONS: Within the limitations of this pilot study, treatment of periodontal-endodontic lesions by performing RCT prior to periodontal treatment was effective, resulting in improvements in clinical parameters together with alveolar bone gains in a majority of teeth. PMID- 16245468 TI - A longitudinal study of the oral health condition of elderly stroke survivors on hospital discharge into the community. AB - AIM: To investigate the oral health of elderly stroke survivors on discharge from hospital into the community after rehabilitation and six months later compared with community dwelling elderly people without stroke. METHOD: The study took place in the rehabilitation unit of a general medical hospital in Hong Kong. It was a longitudinal study involving 43 elderly survivors of mild to moderate stroke about to be discharged from hospital after rehabilitation and a comparison group of 43 community-dwelling elderly people. Verified clinical examination techniques and measures were used to assess dental caries, periodontal disease, oral hygiene, oral candidiasis, prosthetic status/need and functional disability (Barthel Index [BI]). Socio-demographic information was also collected. RESULTS: Stroke survivors had significantly higher plaque and bleeding scores on hospital discharge and after six months compared with the control group (p<0.01). On hospital discharge, the stroke group also had a poorer periodontal condition with more 4-5mm pockets. The mean BI on hospital admission was 71 and 91 on discharge. Stroke survivors with a higher BI on discharge had lower plaque scores after six months. The tooth condition, number of functional tooth pairs, prosthetic status, presence of oral candidiasis and oral yeast carriage were similar between stroke and comparison groups. CONCLUSIONS: Despite significant objective improvement in functional ability after stroke, elderly survivors returning to the community had significantly poorer periodontal health compared with community dwelling elderly without stroke, and the situation did not improve dramatically over six months. The protracted recovery of hand, arm and oral sensori-motor function is the most likely cause. PMID- 16245469 TI - Tobacco control and the dental profession. A survey of the Organisation for Economic Co-operation and Development (OECD) national dental associations. AB - AIM: To explore the current role played by national dental associations (NDAs) in tobacco control and to assess relevant policy initiatives across OECD countries. DESIGN: A self-complete questionnaire survey. SAMPLE: Chief Executive Officers of NDAs of OECD countries. RESULTS: A response rate of 74% was achieved. An overview of tobacco control policies from OECD member states was obtained. Half of the NDAs reported that they had a policy on tobacco control. Furthermore, a minority of OECD countries appear to have tobacco control guidelines that specifically include reference to the role of dental professionals. Dentists are able to prescribe nicotine replacement therapy (NRT) in less than a third of OECD member states. In a small number of OECD countries there was funding available to encourage dentists to become more actively involved in tobacco control activities. Undergraduate training focusing on tobacco control is not universally provided across all OECD countries. CONCLUSIONS: There is an urgent need to put tobacco control initiatives on the oral health policy agenda of NDAs across the OECD. A range of policy opportunities exist to facilitate greater involvement of the dental profession in tobacco control activities. PMID- 16245470 TI - Epidemiology of traumatised primary teeth in the west-Mediterranean region of Turkey. AB - AIM: To investigate the incidence, aetiological factors and results of dental trauma on primary dentition of the children referred to the Suleyman Demirel University, Department of Paedodontics in a two year period. DESIGN: A retrospective study. PARTICIPANTS: Eighty four children (50 boys and 34 girls) with 159 affected deciduous teeth. RESULTS: The incidence of trauma was slightly higher in boys (59.5%). The age of the children ranged from 9 months to 6 years and the peak ages for trauma were 3 years and 5 years of age in respective genders. Falls were the most frequent cause of traumatic dental injuries (29.7%) and luxations were the most common type of trauma with 135 cases (84.9%). The most frequently performed treatment options were examination and follow up only (37.7%). Half of the cases (51.1%) were referred to our clinic in the first week after the trauma occurred. CONCLUSION: The results of this study suggest that a preventive educational programme should be instituted, directed at parents and teachers at nursery schools, in order to inform them about what to do in dental trauma cases and the importance of immediate attendance for dental care. PMID- 16245471 TI - 21st century endodontics. Part 4. AB - The prime objective of this series of papers has been to focus the attention of the reader on the biological principles of root canal treatment. Accurate diagnosis, knowledge of the root canal morphology and the principles of modern preparation techniques are all essential so that appropriate chemical cleaning and disinfection of the entire root canal system may be accomplished. This paper will consider two further aspects of root canal treatment. First, whether the root canal system should be prepared and obturated in a single visit, and, if not, which intervisit medication is appropriate. Secondly, the modern concepts and techniques for obturation of the root canal system. PMID- 16245472 TI - Dentist-patient relationship and quality care 4. Professional information and informed consent. AB - As patients become relatively more knowledgeable on health issues and more aware of their rights, they become more concerned about receiving professional information and making informed choices. A healthy decision-making process can only be achieved through clear information and fair discussions. Thus, rather than signing a form, quality information for patients and the process of informed consent need to be considered as integral elements of a treatment alliance, a fair dialogue, quality oral care and a sign of respect for patients' rights for self-determination. PMID- 16245474 TI - A conceptual framework for teaching research in nursing. AB - Though research is often referred to the lifeblood, hallmark or cornerstone in the development of a profession (Brink, 1996:2), teaching research in nursing is a challenge. The challenge does not just lie in teaching the subject, but in resistance and unwillingness of students to engage in the subject. In the experience of the researcher, registered nurses identify themselves with being a nurse and a caregiver; the role of researcher has never been internalised. The challenge is to achieve the outcome envisaged, namely, nurses who are knowledgeable consumers of research as well as continuous productive scholars in their application of nursing. Research generates knowledge and knowledge is the basis of caring with excellence. Nursing is an art and a science and the science must produce the knowledge upon which the art is based. The purpose of this article is to propose a conceptual framework of how to teach research in order to achieve such a successful outcome. The conceptual framework proposed in this article is based on four pillars, theoretical knowledge of research, scientific writing, psychological support and experiential learning. The importance of the research facilitator, not just as a teacher but also as a positive role model, is also described. PMID- 16245475 TI - Critical thinking of student nurses during clinical accompaniment. AB - The purpose of this study was to investigate the methods of clinical accompaniment used by clinical facilitators in practice. The findings of the study also reflected facilitators' perceptions regarding critical thinking and the facilitation thereof. A quantitative research design was used. A literature study was conducted to identify the methods of accompaniment that facilitate critical thinking. Data was collected by means of a questionnaire developed for that purpose. Making a content-related validity judgment, and involving seven clinical facilitators in an academic institution, ensured the validity of the questionnaire. The results of the study indicated that various clinical methods of accompaniment were used. To a large extent, these methods correlated with those discussed in the literature review. The researcher further concluded that the concepts 'critical thinking' and 'facilitation' were not interpreted correctly by the respondents, and would therefore not be implemented in a proper manner in nursing practice. Furthermore, it seemed evident that tutor-driven learning realised more often than student-driven learning. In this regard, the requirement of outcomes-based education was not satisfied. The researcher is therefore of the opinion that a practical programme for the development of critical thinking skills during clinical accompaniment must be developed within the framework of outcomes-based education. PMID- 16245476 TI - Reproductive health needs and the reproductive health behaviour of the youth in Mangaung in the Free State province: a feasibility study. AB - From the community profiles conducted by nursing students in Mangaung, the following observation was prevalent: an increase in the prevalence of teenage pregnancy together with a high prevalence of HIV infection was demonstrated. The relationship between young people's reproductive behaviour and the prevalence of HIV is well documented. To address this problem, a community -based education programme is required. Developing a successful community-based educational programme that addresses the needs of youth requires that such a programme be based on the needs of such a group. The research question is applicable: How do youth in Mangaung in the Free State province in South Africa perceive their reproductive health needs and their reproductive health behaviour? The aim of this article is to explore and describe the reproductive health needs and the reproductive health behaviour of the youth in Mangaung. A quantitative research design that is descriptive and explorative in nature was conducted. The reproductive health needs and the reproductive health behaviour of the youth were collected by means of a structured questionnaire with open-ended and closed questions. A purposive non - probability sampling method was utilized and (n=250) drawn from the youth. The justification of the sample was further enhanced by collecting data from youth aged 13-25 at three high schools in Mangaung and at the health centre of MUCPP. Qualitative data obtained from open-ended questionnaires was coded and analyzed by using Tech's (1990) content analysis approach. A descriptive statistical analysis was performed on the quantitative data from closed questions. A descriptive analysis of the participant's ages and their perceived reproductive health needs and reproductive health behaviour was done. The mean age of the participants was 18.6, which could be because all the respondents were of school- going age. The results indicated that the youth received insufficient reproductive health information to be able to prevent pregnancy and HIV/AIDS. The special needs of youth were indicated by 92% who requested that the clinic should accommodate both boys and girls and 96% of the respondents requested sessions for discussions facilitated by a professional person. A total of 95% of the youth requested that a special programme of activities be conducted during school holidays. The risk behaviours practised by the youth were: having sex with a stranger, many partners, drinking alcohol, and using drugs and dagga. The researcher recommends that such a programme be supported by different university departments and rolled out to different PMID- 16245477 TI - HIV/AIDS issues in the workplace of nurses. AB - HIV/AIDS is a global problem with an estimated 40 million infected people. In less than two years, this figure will leap to 100 million according to the World Health Organisation (WHO). By 2005, 65 million people will be infected. Half of the number of people in this group will be under 25 years old, and will die before they reach the age of 35. In a South African study done by the Human Science Research Council and published in 2003, regarding the impact of HIV/AIDS on the health sector, the findings were that 15% of health workers in public and private hospitals tested positive for HIV antibodies. Together with these facts above it was found that 46.2 percent of patients served in medical and paediatric wards tested positive for HIV. These factors have major implication for staffing in the future and the role of the nurse manager in South Africa. THE AIM OF THE STUDY: To explore the management of HIV/AIDS in the workplace of nurses in selected health services in KwaZulu-Natal. This research was part of a greater study on the exploration of the presence of caring as part of nursing management. THE METHODOLOGY: The qualitative research approach was used with a phenomenological design, which ensured that the richness and the complexities are reflected in the study. The data was collected by means of an open-ended question to nurse managers during an interview. The first question posed was; How do you or your services care for nurses in this hospital? Secondly nurse managers were asked, To explain their role in caring for HIV/AIDS positive nurses on their staff establishment. THE RESULTS: A qualitative analysis of the interviews with nurse managers indicated that they rate HIV/AIDS issues as an important part of their management task. Four main themes were identified, namely HIV/AIDS, counselling, dying of AIDS and funerals. Rich descriptions of these themes are given in this paper. Nurse managers in the health services are managing HIV/AIDS affected nurses, but are doing so without any formal policy on HIV/AIDS in the workplace. It is recommended that nurse managers deal with HIV/AIDS issues in the workplace by raising HIV/AIDS awareness regarding HIV/AIDS issues and HIV/AIDS in the workplace. CONCLUSIONS: Nurse managers saw their role regarding nurses with HIV/AIDS mainly as supportive--being involved in family problems and counselling and guiding nurses that live with HIV/AIDS. The health services in KwaZulu-Natal, and especially nurses, are currently affected by HIV/AIDS in a significant way. PMID- 16245478 TI - Verbal abuse of nurses by physicians in a private sector setting. AB - The aim of the study was to determine whether physicians in a private sector setting verbally abuse nurses, and to what extent. A review of the literature showed that verbal abuse by physicians accounts for the highest incidence of aggression towards nurses in health care, and that it is strongly related to turnover rates. It has also been reported in recent studies that within the context of verbally abusive episodes, patient care, work productivity, morale and job satisfaction have been negatively affected. Of the 120 questionnaires that were distributed among registered and enrolled nurses, 83 were returned in time to be used, which was a response rate of 69%. This response rate is consistent with previous studies and is exactly the same as for a study that was done in Turkey (Uzun, 2003:81). The questionnaire used was one that has been adapted from the Verbal Abuse Scale developed by Manderino and Berkey (1997:50) and the Revised Conflict Tactics Scale (CTS 2) as described by Little (1999:24), and was in the form of a 4-point Likert-scale with one open ended question. The results showed that 79% of the nurses admitted that verbal abuse was taking place. Forms of direct verbal abuse included 81% that felt they were criticised unjustly, 76% that were screamed at in front of others, and 81% that felt physicians vented their frustration on them. The results obtained in the study were consistent with previous studies done elsewhere and it indicated that nurses still experience high levels of verbal abuse in the workplace. PMID- 16245479 TI - The career goals of nurses in some health care settings in Gauteng. AB - In nursing, purposeful career planning is essential if nurse practitioners want to make the right decisions about their work in order to strive towards and accomplish a meaningful quality of working life. Nurses should identify their career goals to be able to investigate their different career opportunities in their field of interest and direct their work according to a work strategy for years ahead. The purpose of this study was to explore and describe the career goals of post-basic nursing students with the aim of describing management strategies to guide the future career of post-basic nursing students in climbing the career ladder effectively and obtaining their set career goals. An explorative, descriptive, qualitative design was selected where the researcher worked inductively to explore and describe the needs (goals) and future planned actions of the participants regarding their career management as viewed for a period of five years. The researcher purposively and conveniently identified the sample as all the post-basic nursing students, namely 250 students, who were registered for the first, second and third year of nursing management courses in that period at a South African residential university. Two structured, open questions were developed. Each participant received the questions in writing and was asked to answer them. The QSR NUD*IST program was used for the qualitative management (categorization) of data. The results of the research questions related to five categories, namely becoming empowered, being promoted, being educated and professionally developed, partaking in research and taking up new projects. PMID- 16245480 TI - Early intervention care programme for parents of neonates. AB - Parents with neonates in the neonatal intensive care unit (NICU) experience different needs at different stages of their neonates' stay in the NICU. The needs of parents with neonates in NICU's play an important role in aspects such as the ability to cope with changing parental roles and emotions, the relationship between parent and infant and the managing of the parents' own needs. The aim of this study was to develop an intervention care programme for parents with neonates in the NICU. This intervention care programme will empower parents to manage their own needs and the needs of their neonates while the neonate is admitted to the NICU and after discharge from the NICU / hospital. Literature is available on care programmes for neonates, but not on programmes for the parents of neonates in NICU. The study was a multi-phased study, using qualitative methodologies to determine the needs of South African parents with neonates in level III NICU's. In phase I, the needs of parents with neonates in NICU were elicited qualitatively. The needs were identified from the data and the results led to the implementation of phase II. In phase II the question was adjusted and new data was collected. Phase III was implemented to validate the data derived from phases I and II. The data was categorised in different need categories and these categories were used to plan an intervention care programme for parents with neonates in NICU's. The programme provides information to address needs as identified by parents in the research study and as derived from the literature. Need categories identified from the study and literature were as follows: information, communication, emotional, learning, discharge and individual needs. This programme is available in electronic format to enable parents to obtain information according to their changing needs and to provide unlimited access to updated information. The "Early intervention care programme for parents of neonates" will empower parents to manage their own needs and the needs of their neonates while the neonate is admitted to the NICU and after discharge from the NICU / hospital. PMID- 16245481 TI - Virtue ethics: an approach to moral dilemmas in nursing. AB - Nurses are increasingly confronted with situations of moral difficulty, such as not to feed terminally ill patients, whistle blowing, or participation in termination of pregnancy. Most of these moral dilemmas are often analyzed using the principle-based approach which applies the four moral principles of justice, autonomy, beneficence, and non-malificence. In some instances, consequentialism is considered, but these frameworks have their limitations. Their limitations has to do with a consideration for the interpersonal nature of clinical nursing practice on the one hand, and is not always clear on how to judge which consequences are best on the other hand. When principles are in conflict it is not always easy to decide which principle should dominate. Furthermore, these frameworks do not take into account the importance of the interpersonal and emotional element of human experience. On the contrary, decision-making about moral issues in healthcare demands that nurses exercise rational control over emotions. This clearly focuses the attention on the nurse as moral agent and in particular their character. In this article I argue that virtue ethics as an approach, which focus of the character of a person, might provide a more holistic analysis of moral dilemmas in nursing and might facilitate more flexible and creative solutions when combined with other theories of moral decision-making. Advancing this argument, firstly, I provide the central features of virtue ethics. Secondly I describe a story in which a moral dilemma is evident. Lastly I apply virtue ethics as an approach to this moral dilemma and in particular focusing on the virtues inherent in the nurse as moral agent in the story. PMID- 16245483 TI - Musing on Blakemore's paradox. PMID- 16245482 TI - Lesotho midwives' utilization of non-pharmacological pain management methods during the first of stage labour. AB - The purpose of this study was to determine the use of non-pharmacologic methods of pain management used by midwives in Lesotho. The research design was non experimental and of a descriptive nature. The data was obtained by means of a structured questionnaire which was compiled after a thorough literature analysis was done. Midwives, working in the Maternity wards of the Christian Hospital Association of Lesotho as well as the government Hospitals completed the questionnaires. All data was analysed on a nominal descriptive level. According to the results, the midwives indicated that they were taught non-pharmacologic methods of pain management, however they expressed that they inadequately use these methods during the first stage of labour due to shortage of staff, lack of privacy and space, a high midwife-mother ratio, culture and hospital policies. In the light of these findings, recommendations were made of maximizing the use of non-pharmacologic methods during the first stage of labour. PMID- 16245484 TI - Visually perceived eye level and horizontal midline of the body trunk influenced by optic flow. AB - The eye level and the horizontal midline of the body trunk can serve, respectively, as references for judging the vertical and horizontal egocentric directions. We investigated whether the optic-flow pattern, which is the dynamic motion information generated when one moves in the visual world, can be used by the visual system to determine and calibrate these two references. Using a virtual-reality setup to generate the optic-flow pattern, we showed that judged elevation of the eye level and the azimuth of the horizontal midline of the body trunk are biased toward the positional placement of the focus of expansion (FOE) of the optic-flow pattern. Furthermore, for the vertical reference, prolonged viewing of an optic-flow pattern with lowered FOE not only causes a lowered judged eye level after removal of the optic-flow pattern, but also an overestimation of distance in the dark. This is equivalent to a reduction in the judged angular declination of the object after adaptation, indicating that the optic-flow information also plays a role in calibrating the extraretinal signals used to establish the vertical reference. PMID- 16245485 TI - Visual perception of writing and pointing movements. AB - Studies of movement production have shown that the relationship between the amplitude of a movement and its duration varies according to the type of gesture. In the case of pointing movements the duration increases as a function of distance and width of the target (Fitts' law), whereas for writing movements the duration tends to remain constant across changes in trajectory length (isochrony principle). We compared the visual perception of these two categories of movement. The participants judged the speed of a light spot that portrayed the motion of the end-point of a hand-held pen (pointing or writing). For the two types of gesture we used 8 stimulus sizes (from 2.5 cm to 20 cm) and 32 durations (from 0.2 s to 1.75 s). Viewing each combination of size and duration, participants had to indicate whether the movement speed seemed "fast", "slow", or "correct". Results showed that the participants' perceptual preferences were in agreement with the rules of movement production. The stimulus size was more influential in the pointing condition than in the writing condition. We consider that this finding reflects the influence of common representational resources for perceptual judgment and movement production. PMID- 16245487 TI - How configurations of binocular disparity determine whether stereoscopic slant or stereoscopic occlusion is seen. AB - A partially occluded contour and a slanted contour may generate identical binocular horizontal disparities. We investigated conditions promoting an occlusion resolution indicated by an illusory contour in depth along the aligned ends of horizontally disparate line sets. For a set of identical oblique lines with a constant width added to one eye's view, strength, depth, and stability of the illusory contour were poor, whereas for oblique lines of alternating orientations the illusory contours were strong, indicating a reliance on vertical size disparities rather than vertical positional disparities in generating perceived occlusion. For horizontal lines, occlusion was seen when the lines were of different lengths and absolute width disparity was invariant across the set. In all line configurations, when the additional length was on the wrong eye to be attributed to differential occlusion, lines appeared slanted consistent with their individual horizontal disparities. This rules out monocular illusory contours as the determining factor. PMID- 16245486 TI - Confirming statements about pictures of natural scenes: evidence of the processing of gist from eye movements. AB - Combined displays of graphics and text, such as figure captions in newspapers and books, lead to distinctive inspection patterns, or scanpaths. Readers characteristically look very briefly at the picture, and then read the caption, and then look again at the picture. The initial inspection of the picture is the focus of interest in the present experiment, in which we attempted to modify the inspection by giving participants advance knowledge of the subject of a sentence (the cued object) that was to be verified or denied on the basis of whether it correctly described some aspect of the scene shown in the picture. Eye fixations were recorded while the viewers looked at the picture and the sentence in whatever sequence they chose. By allowing viewers to know the subject of the sentence in advance, we asked whether patterns of fixations on the sentence and on the second inspection of the picture would reflect prior knowledge of the focus of the sentence. Providing advance information did not influence eye movements while reading the sentence. It did, however, increase the number of fixations in the initial inspection of the picture, and it also reduced the number and duration of the fixations on the pictures overall. The results suggest that cueing participants to the object allowed increased coding in the initial inspection of the picture, though the benefit of such coding only becomes apparent when the picture is inspected for the second time. PMID- 16245488 TI - Capacity limitation in short-term visual memory for contour curvature. AB - I measured the difference threshold for contour curvature in short-term visual memory (STVM) using a two-interval forced-choice partial discrimination task. In experiments 1 and 2, the study stimulus consisting of 1 to 4 curved contours was briefly presented. It was followed by a single contour stimulus after a retention interval. The subjects judged if the test stimulus had a higher or lower curvature than the corresponding study contour. The results of experiment 1 showed that the Weber fraction increased monotonically with increasing set size. The results of experiment 2 clarified that the set-size effect was not due to a temporal limitation in encoding resulting from the short exposure time. In experiment 3, the study stimuli always consisted of 4 items, but the numbers of memorised items were different in each condition. Nevertheless, the results showed the set-size effect, which indicated that its occurrence depended largely on the capacity limitation in short-term visual memory (STVM) storage. Otherwise, the Weber fraction was not hugely higher for set size 4 compared with set size 1. It was concluded that only 1 object could be retained in STVM with high fidelity, but that at least 4 objects could be retained in STVM with some degree of fidelity. PMID- 16245489 TI - When feature information comes first! Early processing of inverted faces. AB - We investigated the early stages of face recognition and the role of featural and holistic face information. We exploited the fact that, on inversion, the alienating disorientation of the eyes and mouth in thatcherised faces is hardly detectable. This effect allows featural and holistic information to be dissociated and was used to test specific face-processing hypotheses. In inverted thatcherised faces, the cardinal features are already correctly oriented, whereas in undistorted faces, the whole Gestalt is coherent but all information is disoriented. Experiment 1 and experiment 3 revealed that, for inverted faces, featural information processing precedes holistic information. Moreover, the processing of contextual information is necessary to process local featural information within a short presentation time (26 ms). Furthermore, for upright faces, holistic information seems to be available faster than for inverted faces (experiment 2). These differences in processing inverted and upright faces presumably cause the differential importance of featural and holistic information for inverted and upright faces. PMID- 16245490 TI - Central visual persistences: I. Visual and kinesthetic interactions. AB - Phenomena associated with 'central visual persistences' (CPs) are new to both medical and psychological literature. Five subjects have reported similar CPs: positive afterimages following brief fixation of high-contrast objects or drawings and eye closure. CPs duplicate shapes and colors of single objects, lasting for about 15 s. Unlike retinal afterimages, CPs do not move with the eyes but are stable in extrapersonal space during head or body rotations. CPs may reflect sustained neural activity in neurons of association cortex, which mediate object perception. A remarkable finding is that CPs can be moved in any direction by the (unseen) hand holding the original seen object. Moreover, a CP once formed will 'jump' into an extended hand and 'stick' in that hand as it moves about. The apparent size of a CP of a single object is determined by the size of the gap between finger and thumb, even when no object is touched. These CPs can be either magnified or minified via the grip of the extended hand. The felt orientation of the hand-held object will also determine the orientation of the CP seen in that hand. Thus, kinesthetic signals from hand and arm movements can determine perceived location, size, and orientation of CPs. A neural model based on physiological studies of premotor, temporal, parietal, and prefrontal cortices is proposed to account for these novel phenomena. PMID- 16245491 TI - Interaction of vision and movement via a mirror. AB - I have noticed a striking effect that vision can have on movement: when a person makes circular motions with both hands, clockwise with the left hand, counterclockwise with the right hand, while watching the reflection of one hand in a parasagitally placed mirror, if one arm makes a vertical excursion, the other arm tends to make the same vertical excursion, but not typically if the excursing arm is viewed in plain vision. This observation may help in understanding how visual feedback via a mirror may be beneficial for rehabilitation of some patients with movement deficits secondary to certain neurologic conditions, and illustrates that the traditional division of neural processes into sensory input and motor output is somewhat arbitrary. PMID- 16245492 TI - Magic and fixation: now you don't see it, now you do. PMID- 16245493 TI - Recommendations for the management of prosthetic valve thrombosis. AB - Prosthetic valve thrombosis (PVT) is a life-threatening disease, for which treatment strategies have been controversial. Herein, existing data on management options are reviewed, and conclusions drawn as to the choice and use of treatment strategies for PVT. The use of transesophageal echocardiography (TEE) allows distinction to be made between obstructive and non-obstructive PVT by the presence or absence of occluder motion limitation. The differentiation of PVT from pannus and vegetation is, however, still limited by TEE. The incidence of PVT has been underestimated by not taking into account a large percentage of non obstructive PVT. Although the standard treatment for PVT has been surgery, thrombolysis has lower mortality rates, particularly in patients in NYHA functional classes III-IV. The lowest complication rates with thrombolysis have been achieved in patients with non-obstructive PVT. Pregnancy, left atrial appendage thrombi and large PVT are not contraindications to thrombolysis. The third therapeutic option is anticoagulant therapy. The detrimental effect of anticoagulant treatment in obstructive PVT was shown in a prospective study. Non obstructive thrombi of > 5 mm length have been treated with higher success rates and lower complication rates by thrombolysis than by anticoagulant treatment. In conclusion, all patients with suspected PVT should undergo multiplane TEE. Thrombolysis is the first-line treatment for obstructive PVT, independent of NYHA class and thrombus size if there are no contraindications. Serial TEE studies must be conducted during thrombolysis. Surgery should be reserved for those patients in whom thrombolysis is contraindicated, or has failed. Initial anticoagulant therapy is recommended only for small, non-obstructive PVT if anticoagulation had been subtherapeutic; otherwise, thrombolysis is the treatment of choice if there are no contraindications. PMID- 16245494 TI - An audit of anticoagulation practice among UK cardiothoracic consultant surgeons following valve replacement/repair. AB - BACKGROUND AND AIM OF THE STUDY: In 1998, the American College of Cardiology and The American Heart Association (ACC/AHA) published guidelines for the postoperative anticoagulation of patients who have undergone heart valve replacement. The American College of Chest Physicians made similar recommendations in 2001. The present survey was conducted to review anticoagulation practice among UK consultant cardiac surgeons, and to assess compliance with these guidelines. METHODS: An anonymous postal questionnaire was distributed to 185 adult cardiac surgeons identified from the Society of Cardiothoracic Surgeons of Great Britain and Ireland (SCTS). RESULTS: The analysis was based upon 97 replies. All consultants use lifelong warfarin after mechanical valve replacement. In general, target INR ranges were lower for aortic valves compared with mitral valves. Some 53% (51/97) of consultants never use warfarin after bioprosthetic aortic valve replacement (AVR), compared with 33% (28/86) after bioprosthetic mitral valve replacement (MVR). Temporary (< or = 3 months) warfarin is used by 47% (46/97) of consultants after bioprosthetic AVR and by 63% (54/86) after bioprosthetic MVR. Some 64% (52/81) of consultants use warfarin after mitral valve repair, when an annuloplasty ring is inserted. This was always temporary (< or = 6 months). Aspirin is used long term by 54% (44/82) of consultants after mitral valve repair. CONCLUSION: All consultant cardiac surgeons adequately anticoagulate their patients after mechanical valve replacement. Only 16% (16/97) of cardiac surgeons follow current guidelines for the postoperative anticoagulation of bioprosthetic AVR. Only 28% (24/86) of consultant cardiac surgeons comply with guidelines for bioprosthetic MVR. No guidelines exist for the anticoagulation of patients after mitral valve repair. Guidelines need to be reviewed for the anticoagulation of patients undergoing bioprosthetic valve replacement and formulated for patients undergoing mitral valve repair. PMID- 16245495 TI - Mitral valve re-repair. AB - BACKGROUND AND AIM OF THE STUDY: Between January 1998 and March 2004, a total of 1,596 mitral valve repairs was performed at the authors' institution. Among these operations, 23 constituted re-repair of previously repaired mitral valves. The cases were analyzed retrospectively to ascertain the etiology of the failed repair, the technical considerations of re-repair, and outcome. METHODS AND RESULTS: Previous repairs were performed for myxomatous valve disease in 14 of 23 patients. In seven patients recurrence occurred within three months of the original procedure (two as a result of infective endocarditis and five after technical failure). Incorrect annuloplasty ring size or failure to use a ring, resulting in progressive annular dilatation with or without new valvular dysfunction, were the main causes of delayed recurrence of mitral regurgitation (i.e. more than one year after the original repair procedure). Most patients (n = 12) required a new annuloplasty ring and additional repair procedures. Two patients died within the immediate post-operative period, and five died subsequently from non-cardiac causes during a mean follow up period of 24 months. Among the remaining 16 patients, 13 were free from mitral regurgitation at follow up and two required subsequent mitral valve replacement. CONCLUSION: Re-repair of the mitral valve is a technically feasible operation that can be safely performed with good functional results. It is, however, associated with some increase in postoperative morbidity and mortality due to the reoperative setting. PMID- 16245496 TI - Mechanism of hemolysis after mitral valve repair and new surgical management: prosthetic annuloplasty ring covered with autologous pericardium. AB - BACKGROUND AND AIMS OF THE STUDY: Hemolysis after mitral valve repair is a rare occurrence, but is one of the complications leading to reoperation. Since 1999, mitral valve repair at the authors' institution has been performed using a prosthetic annuloplasty ring covered with autologous pericardium to prevent this complication. The study aims were to investigate the mechanism of hemolysis after mitral valve repair and to describe the surgical management of this complication. METHODS: This retrospective study comprised 204 consecutive patients who underwent mitral valve repair using an annuloplasty ring between October 1991 and April 2000 at the authors' institution. Patients were allocated to the non covered ring group (n = 174) and the covered ring group (n = 30), and compared for the degree of mitral regurgitation (MR), serum levels of lactate dehydrogenase (LDH), and occurrence of hemolysis. The degree and flow pattern of MR, and patient prognoses were described for hemolytic patients. RESULTS: Postoperative MR and serum LDH were not significantly high in either group. A total of seven patients presented with hemolysis; postoperative echocardiography revealed MR to be mild in two patients, moderate in three and severe in two. Collision of the regurgitant jet into the artificial ring was evident in all seven patients. A beta-blocker proved effective in treating hemolysis in three patients, mitral re-repair was performed in three, and a prosthetic mitral valve was inserted in one patient. None of the patients in the covered ring group presented with hemolysis. CONCLUSION: The major cause of hemolysis after mitral repair was collision of the regurgitant jet into the artificial ring. The simple technique used herein prevented contact of the regurgitant jet with the rough surface of the ring, and may in turn have prevented hemolysis. In selected patients, hemolysis was improved by beta-blocker administration. PMID- 16245497 TI - Effect of cutting second-order chordae on in-vivo anterior mitral leaflet compound curvature. AB - BACKGROUND AND AIM OF THE STUDY: Leaflet curvature determines leaflet stress. In order to assess the influence of second-order chordae (2 degrees CT) on anterior mitral valve leaflet (AMVL) geometry, AMVL curvature was measured before (Baseline) and after (CUT) cutting the 2 degrees CT. METHODS: Miniature radiopaque markers were sutured onto the AMVL in eight sheep: four along the central-meridian from mid-septal annulus to the free-margin; and one each at the 2 degrees CT insertion. Biplane videofluoroscopic data were acquired (open-chest) before and after CUT. Marker-triplet 3-D coordinates were used to calculate radii of-curvature at LVPmax along the central-meridian (ROCm) and across the AMVL belly (commissure-commissure axis, ROCc-c). RESULTS: CUT did not change LVPmax (111 +/- 12 versus 106 +/- 11 mmHg; p = 0.19). At baseline, the AMVL central meridian had compound curvature: Convex to the left ventricle near the annulus ( ROCm) and concave near the free-margin (+ROCm). After CUT, the AMVL flattened: ROCm increased near the annulus (from -1.37 +/- 0.52 to -12.58 +/- 29.04 cm; p = 0.02), but did not change near the edge. In the commissure-commissure axis, ROCc c was concave to the left ventricle at baseline and increased after CUT in all eight animals. In five sheep, ROCc-c was increased (from 1.93 +/- 1.01 to 2.80 +/ 1.36 cm; p = 0.03), but in three sheep ROCc-c was increased and inverted (from 3.65 +/- 2.17 to -1.72 +/- 0.53 cm; p = 0.03), becoming convex to the left ventricle. CONCLUSION: Compound curvature along the AMVL central-meridian appears to be an intrinsic leaflet property that persists even without support from second-order chordae, whereas concave curvature in the commissure-commissure axis is more dependent on intact second-order chordae. Leaflet compound curvature must be incorporated into future finite element models to characterize leaflet stresses accurately. The importance of second-order chordae in maintaining leaflet shape must be considered during mitral repair. A larger ROC increases leaflet stresses, while reversal of ROC changes tensile stress to compressive stress; this might trigger deleterious leaflet remodeling after chordal cutting. PMID- 16245498 TI - The role of Chordae tendineae in mitral valve competence. AB - BACKGROUND AND AIM OF THE STUDY: The study aim was to understand the role of different mitral valve chordae tendineae, and how damage to them affects valve competence. METHODS: A test apparatus was used to apply pressure to porcine mitral heart valves that were intact and have had selected chords severed. Anterior leaflet strut and marginal chords were selectively severed, as were posterior leaflet basal and marginal chords. Commissural chords were also severed. RESULTS: Severing anterior leaflet marginal chords (p = 0.018) and commissural chords (p = 0.018) significantly reduced mitral valve competence. Severing posterior leaflet marginal and basal chords, and anterior leaflet strut chords, had no significant effect in reducing the pressures that the valves could withstand. Severing a mixture of posterior leaflet basal and marginal chords significantly reduced the pressure withstood by the valves (p = 0.004). CONCLUSION: The study results confirmed that anterior leaflet marginal chords, but not strut chords, are vital for valve competence. Commissural chords were also shown to be vital for mitral valve competence. Several posterior leaflet chords had to be severed to affect mitral valve competence. PMID- 16245499 TI - Are there surgical implications to aortic root motion? AB - BACKGROUND AND AIM OF THE STUDY: By increasing the longitudinal stress in the ascending aorta, downward movement of the aortic root might promote the proximal transverse tears seen in aortic dissections. The study aim was to evaluate the influence of five common cardiac conditions on the magnitude of aortic root displacement in cardiac patients. METHODS: Aortic root contrast injections were analyzed in 90 patients (mean age 68 years) to measure downward motion of the root perpendicular to the plane of the sinotubular junction (STJ). RESULTS: Displacement of the aortic root ranged from 0 to 14 mm (mean 4.8 mm). Patients with aortic insufficiency (AI) showed increased aortic root movement (7.3 versus 4.3 mm, p = 0.003), whereas those with left ventricular hypokinesis (3.7 versus 5.5 mm, p = 0.014) or with myocardial hypertrophy (3.8 versus 5.1 mm, p = 0.073) exhibited reduced downward movement. These variables were independent, and correlated with the magnitude of aortic root motion. A stress analysis of the aortic root, arch and branches of the arch determined that the longitudinal stress approximately 2 cm above the STJ, in the outer curve of the aorta, was increased by 32% in patients with AI compared to patients without AI. CONCLUSION: Patients with cardiac conditions associated with increased aortic root motion such as AI may be at greater risk of aortic dissection because of increased longitudinal stress in the ascending aorta. Therefore, AI should be used as an indicator and aortic root displacement monitored to prevent the risk of aortic dissection. PMID- 16245500 TI - Current status of the Contegra conduit for pediatric right ventricular outflow tract reconstruction. AB - Reconstruction of the right ventricular outflow tract (RVOT) is needed in a wide variety of congenital heart diseases at the time of primary repair, or later for replacement of existing valves or conduits. Ideally, the conduit or valve needed for such reconstruction should be formed of autologous tissue that grows, resists infection, lasts for the life span of the patient, and is readily available in all sizes. Such a conduit is not available, though several alternatives have been used, none of which is without potential drawbacks. One alternative--the Contegra bovine jugular vein conduit (Medtronic, Inc., Minneapolis, MN, USA)--was introduced in 1999 and has gained widespread application, with increasing enthusiasm for its use. The Contegra conduit consists of a bovine jugular vein with an incorporated trileaflet valve. The conduit tissue is extremely pliable and offers optimal conditions for surgical handling. Moreover, the proximal tubular segment allows construction of the proximal anastomosis to the right ventricle, without the use of additional material. Increasingly, experience with the Contegra conduit is being published; hence, a literature search was conducted to evaluate available evidence on current use of the device in pediatric RVOT reconstruction. PMID- 16245501 TI - Aortic valve replacement with new-generation stentless pericardial valves: short term clinical and hemodynamic results. AB - BACKGROUND AND AIM OF THE STUDY: Aortic valve replacement (AVR) with stentless bioprostheses offers superior hemodynamics. In order to overcome the disadvantages of older, stentless valves, a new generation of pericardial stentless prostheses has been developed. Herein, the hemodynamic and clinical results of these substitutes have been evaluated. METHODS: Between March 2002 and May 2004, 85 patients (59 females, 26 males; mean age 73.6 +/- 6.1 years) who underwent AVR received either a bovine (Sorin Pericarbon Freedom; SPF; n = 50) or an equine (3F Aortic Bioprosthesis; 3F; n = 35) pericardial stentless valve. Patients were followed up prospectively at six months after surgery by clinical and echocardiographic examination. The mean follow up period was 5.6 +/- 0.8 months, and was 96.4% complete. RESULTS: Mortality was 2.4% at 30 days (two SPF patients; one died at reoperation for suspected valve thrombosis and one was a non-valve-related death) and 2.5% at follow up (two SPF patients; both nonvalve related). Neither structural valve failure nor endocarditis were observed. Preoperatively, there were no differences in baseline data, functional status and hemodynamics between SPF and 3F patients. The aortic cross-clamp time was similar in both groups (51.7 +/- 11.2 min for SPF; 51.6 +/- 8.2 min for 3F). NYHA functional status improvement was similar in each group (1.8 +/- 0.5 for SPF; 1.7 +/- 0.6 for 3F). The mean transaortic pressure gradient (deltapmean) was reduced in all patients during follow up. With SPF, a lower deltapmean was found for smaller aortic roots (indexed annular diameter (IAD) < 14 mm/m2) as well as in larger (IAD > 14 mm/m2) aortic roots: 8.0 +/- 4.5 mmHg versus 13.2 +/- 7.2 mmHg (p < 0.05) and 6.8 +/- 3.0 mmHg versus 12.8 +/- 4.8 mmHg (p < 0.05), respectively. CONCLUSION: New-generation pericardial stentless aortic valves are very pliable, which facilitates their implantation. Clinical and hemodynamic results with these prostheses are promising. The SPF prosthesis demonstrates excellent performance, and may be superior when implanted in small aortic roots. PMID- 16245502 TI - Mid-term results of freestyle aortic stentless bioprosthetic valve: clinical impact of quantitative analysis of in-vivo three-dimensional flow velocity profile by magnetic resonance imaging. AB - BACKGROUND AND AIM OF THE STUDY: The study aim was to investigate the in-vivo flow profiles of a stentless aortic bioprosthetic valve by MRI flow quantification, and to identify the clinical implication of prosthesis size and implantation method. METHODS: Twenty-six patients with a Freestyle stentless aortic bioprosthetic valve were studied using three-dimensional flow velocity profile by MRI, and compared with four patients with a stented aortic bioprosthetic valve and four healthy volunteers. Flow velocity profiles were analyzed quantitatively by the hydromechanics parameter, mean to peak velocity ratio at peak systole and compared with parameters monitored echocardiographically. RESULTS: In larger-sized valves or full root implantation, flow profiles showed an optimal pattern with low gradients, a high mean to peak velocity ratio, and minimum disturbance which approximated that of a normal valve. By contrast, a subset of patients, notably with 21 mm valves and subcoronary implantation, showed suboptimal flow pattern with high gradient and low mean to peak velocity ratio which approximated that of stented valves. The mean to peak velocity ratio was more strongly related to peak velocity than to the indexed effective orifice area. CONCLUSION: Although stentless aortic bioprostheses have excellent hemodynamic performance, some patients show suboptimal results. This seems to occur more often when the subcoronary technique is used, and especially with 21-mm valves. Care must be taken when using the subcoronary technique with a 21-mm valve in patients with a small body surface area. PMID- 16245503 TI - Three years' experience with the on-x conform-X bileaflet prosthesis for 'atrialized' mitral valve replacement: a preliminary report. AB - BACKGROUND AND AIM OF THE STUDY: Positioning of a mechanical prosthesis outside the native annulus facilitates mitral valve replacement, especially when the annulus is small and calcified, and preservation of the posterior leaflet (PML) is desired. Herein is described the authors' initial experience with a new mechanical bileaflet prosthesis comprising a sheltered leaflet housing and a novel, modified asymmetrical sewing ring for an 'atrialized' implantation technique. METHODS: Forty-seven patients (24 males, 23 females; mean age 65.8 +/- 10.9 years) were operated on for isolated mitral regurgitation or combined stenosis and insufficiency. Ten of the patients underwent valve replacement due to active endocarditis. Preoperative cardiac insufficiency was reflected by a mean NYHA class of 2.6 +/- 0.7. The On-X Conform-X bileaflet mitral valve prosthesis (25/33 mm) was implanted in either an epiannular (43 patients) or intra-annular (four patients) fashion, and the PML and its chordae tendineae were preservable in 33 patients (70.2%). All patients were monitored intraoperatively by transesophageal echocardiography (TEE) and post-operatively by transthoracic echocardiography (TTE). RESULTS: TEE and TTE exhibited excellent function with low mean transvalvular gradients early after surgery (4.9 +/- 2.7 mmHg) and after three months (4.8 +/- 1.4 mmHg). Paravalvular leakage was not detected. Initial postoperative left ventricular (LV) function was almost identical to preoperative findings (ejection fraction: preop. 55.1 +/- 13.7%, early postop. 53.7 +/- 13.6%) and improved slightly after three months (60.1 +/- 6.4%). Clinically significant hemolysis was not apparent (LDH at postoperative day 7: 5.3 +/- 0.8 micromol/l x s). Two patients died from multiorgan failure (4.2%) and four are currently in a reduced condition requiring long-term hospitalization (morbidity 8.5%). After three months all other patients were in an excellent clinical state (mean NYHA class 1.5 +/- 0.6) and being followed up as outpatients. CONCLUSION: Current designs of mechanical bileaflet valves require redundant space in the LV cavity for undisturbed leaflet movement. A small annulus, marked fibrosis and calcification of the valvular apparatus can lead to an impedance of prosthetic leaflet motion. Consequently, the native leaflets are often removed. The cylindrical housing of the On-X valve shelters almost the entire motion of the leaflets, allowing an undisturbed function and improved transvalvular flow pattern. The newly developed asymmetrical sewing ring facilitates anchoring of the ring in an 'atrialized' fashion, while the flexibility of the cuff adapts to all native ring diameters larger than 25 mm. PMID- 16245504 TI - In-vivo turbulent stresses of bileaflet prosthesis leakage jets. AB - BACKGROUND AND AIM OF THE STUDY: Previous studies of leakage jet turbulence have been carried out in vitro, using a Newtonian fluid to simulate blood and large, rigid approximations to the chambers of the heart. The study aim was to apply an in-vivo method of quantifying leakage jet turbulence to a variety of bileaflet mechanical heart valves, and thereafter to determine the effects of exercise and valve design on turbulent shear stresses within leakage flow. METHODS: Bileaflet prostheses sewn to a manual traversing device were implanted in the mitral position of 29 pigs of body weight ca. 90 kg. Pulsed Doppler ultrasound was used to acquire velocity measurements within the leakage jets detected 1 mm upstream of the housing. Analytical techniques were used to estimate peak velocities and maximum turbulent shear stresses from these velocity measurements. RESULTS: Maximum turbulent shear stress was found to rise with increasing ventricular pressure. No leakage turbulence was found from a valve with relatively small leakage gap widths. The Medtronic Parallel valve was found to have considerable significant leakage flow disturbance, even under low ventricular pressure conditions. Similar maximum turbulent shear stress magnitudes were estimated in the leakage jets of the St. Jude Medical, CarboMedics and Sorin Bicarbon valves at medium ventricular pressure conditions. The maximum turbulent shear stresses estimated in these experiments were lower than those found in previous in-vitro measurements. CONCLUSION: Exercise raises the turbulent shear stresses of leakage flow substantially. Hinge design and leakage gap width also affect the magnitudes of these stresses. Leakage flow turbulence may be less damaging to the blood than was previously thought, and is considerably less damaging than forward-flow turbulence. PMID- 16245505 TI - Digital frequency analysis of valve sound phenomena in patients after prosthetic valve surgery: its capability as a true home monitoring of valve function. AB - BACKGROUND AND AIM OF THE STUDY: Depending on the individual risk profile of a patient, disturbances of the functional integrity of mechanical heart valve prostheses occur in up to 2.5% of patients each year. The early phase of prosthetic dysfunction (due to thrombus formation, tissue ingrowth or endocarditis) usually remains undiagnosed, as patients do not present with symptoms in this situation, and imaging techniques (echocardiography, fluoroscopy) demonstrate normal occluder motion. The delay between the onset of prosthetic valve dysfunction and its clinical manifestation may result in complications (e.g. thromboembolism) or extended therapeutic options (e.g. reoperation rather than more intensive anticoagulation). METHODS: A total of 291 patients with mechanical heart valves was allocated to four different sub-groups, and each measured their valve sounds regularly with the 'ThromboCheck' device. Depending on the subgroup, the signals were compared with different reference signals. Patients in whom a suspicious signal was detected were immediately contacted and examined meticulously. RESULTS: Fourteen patients were found to have suspicious signals. In 13 patients, valve dysfunction was confirmed by fluoroscopy, but in four cases neither transthoracic nor transesophageal echocardiography detected abnormal occluder motion or 'musses' adjacent to the prosthesis. Normal valve sounds returned in four patients who underwent thrombolytic therapy. All patients regularly recorded and passed on their signals. Surveys revealed high acceptance and easy handling of the Thrombocheck device. CONCLUSION: Home monitoring of sound pressure measurements of prosthetic valves by digital frequency analysis via a Fast Fourier transformation may detect even very mild alterations of prosthetic valve function. The next evolution of control systems, allowing for registration of flow, frequency spectrum and electrocardiography, opens potential applications for Internet-based, remote monitoring of cardiac patients. PMID- 16245506 TI - Cinefluoroscopic assessment of mechanical disc prostheses: its value as a complementary method to echocardiography. AB - BACKGROUND AND AIM OF THE STUDY: Many types of mechanical prostheses are used for heart valve replacement, but it is difficult to distinguish between them using transthoracic echocardiography. Hence, cinefluoroscopy complements the echocardiographic evaluation of cardiac prostheses. The aims of the present study were to: (i) describe the contribution of cinefluoroscopy in identifying different prostheses; (ii) compare gradients obtained by Doppler echocardiography with the opening angle of the discs assessed by cinefluoroscopy; and (iii) assess the ability of cinefluoroscopy to distinguish normal from dysfunctional prostheses. METHODS: A total of 229 mechanical disc prostheses was prospectively evaluated with cinefluoroscopy. Eight prosthetic valves (six aortic, two mitral) were excluded due to the coexistence of severe left ventricular dysfunction. Thus, the final analysis comprised 221 prosthetic valves (146 aortic, 75 mitral). RESULTS: Based on the characteristics of the ring and the discs, cinefluoroscopy identified 87 single-leaflet and 134 bileaflet prostheses. Disc motion allowed distinction to be made between normal and dysfunctional prostheses (opening angle: 74 +/- 13 degrees versus 49 +/- 18 degrees). Fluoroscopy could not define disc profile or the ring in 6% of aortic valves and in 26% of mitral prostheses. The technique could be used to identify the TriTechnologies and HP-Biplus valves, but could not provide data on prosthetic function due to radiolucency of the discs. Among the 146 aortic prostheses, Doppler echocardiography helped to identify 109 normal valves and 37 dysfunctional valves. Among 75 mitral prostheses, 54 normal and 21 dysfunctional prosthetic valves were identified. When both methods were correlated, the sensitivity, specificity and positive and negative predictive values of fluoroscopy to distinguish normal from malfunctioning prostheses were 83%, 80%, 89%, and 71%, respectively. CONCLUSION: Each prosthesis type has radioscopic characteristics that allow its identification. Fluoroscopy permitted rapid and easy evaluation of mechanical prosthetic valve function, and in most cases allowed a distinction to be made between normal and dysfunctional prostheses. The presence of high gradients by Doppler echocardiography, with normal opening angles by fluoroscopy, and without pannus on transesophageal echocardiography, is indicative of patient-prosthesis mismatch. Fluoroscopy was superior to echocardiography in identifying disc motion, whilst Doppler study allowed the measurement of gradients and areas, and semiquantification of regurgitation. Thus, cinefluooscopy rapidly provides valuable information which is complementary to that obtained by echocardiography. PMID- 16245507 TI - Bone marrow-derived myofibroblasts are present in adult human heart valves. AB - BACKGROUND AND AIM OF THE STUDY: Endothelial, smooth muscle and cardiomyocyte chimerism has been shown to occur in the human heart. It is currently unknown whether the bone marrow contributes to cellular components of adult human heart valves. Here, it was determined whether bone marrow-derived smooth muscle-like cells (SMLC) are present in the heart valves of adult subjects. METHODS: By combining immunofluorescence staining and fluorescence in-situ hybridization (FISH) for X and Y chromosomes, the heart valves of gender-mismatched bone marrow transplant patients were examined for the presence of chimeric cells expressing calponin, a smooth muscle-specific protein. Concomitant staining for CD68 antigen was carried out to exclude cells of a monocytic lineage. RESULTS: The mean percentage of bone marrow-derived SMLC in valves was 0.28 +/- 0.03%, with the total proportion of chimeric cells estimated at 0.71 +/- 0.05%. The mean proportion of CD68+ cells was 0.33 +/- 0.05%. Not a single cell stained doubly for calponin and CD68 antigen. CONCLUSION: These data establish, for the first time, human bone marrow as a source of progenitor cells contributing to SMLC in adult human heart valves. PMID- 16245508 TI - Mesostructures of the aortic valve. AB - BACKGROUND AND AIM OF THE STUDY: The aortic valve cusp is commonly described as a three-layered structure containing circumferentially aligned fiber bundles. Little is known, however, regarding fiber bundle sizes, branching patterns, or how they are connected. This is because previous morphological studies relied primarily on histological sectioning and staining techniques, which tend to affect all of the collagen, regardless of structure or orientation. METHODS: To address this problem, a novel system was developed for the visualization and analysis of the intermediate-scale 'mesostructures' of aortic valve cusps. Mesostructures are defined as the branching fiber bundle and membrane structures that make up the valve. This system uses elliptically polarized light to provide contrast between collagen mesostructures without the need for embedding, staining, or other contrast-enhancing techniques. Using this system, high resolution images of 42 whole porcine aortic valve cusps were acquired in an unloaded (i.e. resting) condition and during application of controlled manipulation. Image-processing algorithms were developed to quantify fiber bundle morphological features and produce detailed maps of the fiber bundle patterns. RESULTS: Fiber bundle sizes and patterns were found to be significantly different for each of the three cusps. The non-coronary cusp had a significantly smaller bundle diameter (0.9 +/- 0.07 mm) than the left and right coronary cusps (1.1 +/- 0.08 mm). The left and non-coronary cusps appeared to be mirror images of each other, whereas the right coronary cusp was self-symmetric. When applying controlled loads to the cusp specimens, thin, overlapping, collagenous membranes were often found which connected the fiber bundles. Interesting pinnate fiber branching patterns were also found. CONCLUSION: These morphological results were strikingly different than the currently accepted three-layer description, and may provide valuable insight into aortic valve structure-function relationships. PMID- 16245509 TI - Successful thrombolytic therapy for prosthetic mitral valve thrombosis with hemodynamic obstruction: case report and review of the literature. AB - Prosthetic valve thrombosis (PVT) is a serious complication of mechanical valves. The use of thrombolytic therapy to relieve thrombotic prosthetic valve obstruction is accepted for right-sided valves, but its role in left-sided valves and hemodynamically stable patients awaits clarification. The case is presented of a patient in whom prosthetic heart valve thrombosis was treated successfully with streptokinase under transesophageal echocardiographic guidance. PMID- 16245510 TI - A novel Helcococcus-like organism causing endocarditis in an injecting drug user. AB - A case is presented of infective endocarditis caused by a previously undescribed Helcococcus-like organism, in a patient with human immunodeficiency virus infection. The organism, which could not be identified phenotypically in the routine laboratory, was identified using partial 16S rRNA gene sequencing. PMID- 16245511 TI - Valve-sparing root replacement after prior Starr-Edwards aortic valve replacement. AB - Historically, patients with prior aortic valve replacements who subsequently present with an ascending aortic aneurysm require placement of a valve conduit. However, if the patient has a functional mechanical valve with proven long-term durability, an attempt can be made to preserve the intact valve and to graft the aneurysmal aortic root. The case is described of a patient with a previously placed Starr-Edwards aortic valve who subsequently developed a 6-cm ascending aortic aneurysm. By removing the valve ball and using the existing sewing ring, a proximal graft anastomosis was created with ease, eliminating valve excision. PMID- 16245512 TI - Iatrogenic left atrioventricular fistula after aortic valve re-replacement. AB - Left ventriculoatrial fistula is a very rare cardiac disorder. The case is reported of a male patient who was admitted to the authors' clinic with dyspnea and fatigue. The patient had an atrioventricular fistula between the left atrium and left ventricle, and had undergone aortic valve re-replacement for prosthesis malfunction about one year previously. The fistula was considered to be the result of iatrogenic injury related to valve re-replacement surgery. PMID- 16245513 TI - Libman-sacks endocarditis and primary antiphospholipid syndrome. AB - Cardiac involvement is a not uncommon complication in patients with antiphospholipid syndrome (APS). Herein, the case is reported of cardiac failure in a female patient with Libman-Sacks endocarditis and with primary APS diagnosed eight years previously. Aggressive anticoagulation therapy and medical treatment for the cardiac failure over a 12-month period resulted in a partial regression of the severe mitral regurgitation. Close clinical and echocardiographic surveillance during the follow up of patients with APS and heart valve disease is mandatory. Optimal treatment, including adequate aggressive anticoagulation therapy and specific treatment for heart failure, may play a pivotal role in reducing the severity of valve dysfunction in these patients. PMID- 16245514 TI - A quadricuspid aortic valve in a patient with neurofibromatosis type 1. AB - Quadricuspid aortic valve is a rare cardiovascular abnormality. Herein is described the case of a male patient with neurofibromatosis type 1 (NF1) who was found to have a quadricuspid aortic valve causing severe aortic regurgitation. Although congenital cardiac malformations have been described in patients with NF1, to the best of the present authors' knowledge this is the first time that a quadricuspid valve in a patient with neurofibromatosis has been described. PMID- 16245515 TI - Decellularization of bovine pericardium for tissue-engineering by targeted removal of xenoantigens. PMID- 16245516 TI - Zinc oxide nanostructures: synthesis and properties. AB - This article provides a comprehensive review of the current research activities that focus on the ZnO nanostructure materials and their physical property characterizations. It begins with the synthetic methods that have been exploited to grow ZnO nanostructures. A range of remarkable characteristics are then presented, organized into sections describing the mechanical, electrical, optical, magnetic, and chemical sensing properties. These studies constitute the basis for developing versatile applications of ZnO nanostructures. PMID- 16245518 TI - Mechanical properties of carbon nanotubes and their polymer nanocomposites. AB - More than 10 years have passed since carbon nanotubes (CNT) have been found during observations by transmission electron microscopy (TEM). Since then, one of the major applications of the CNT is the reinforcements of plastics in processing composite materials, because it was found by experiments that CNT possessed splendid mechanical properties. Various experimental methods are conducted in order to understand the mechanical properties of varieties of CNT and CNT-based composite materials. The systematized data of the past research results of CNT and their nanocomposites are extremely useful to improve processing and design criteria for new nanocomposites in further studies. Before the CNT observations, vapor grown carbon fibers (VGCF) were already utilized for composite applications, although there have been only few experimental data about the mechanical properties of VGCF. The structure of VGCF is similar to that of multi wall carbon nanotubes (MWCNT), and the major benefit of VGCF is less commercial price. Therefore, this review article overviews the experimental results regarding the various mechanical properties of CNT, VGCF, and their polymer nanocomposites. The experimental methods and results to measure the elastic modulus and strength of CNT and VGCF are first discussed in this article. Secondly, the different surface chemical modifications for CNT and VGCF are reviewed, because the surface chemical modifications play an important role for polymer nanocomposite processing and properties. Thirdly, fracture and fatigue properties of CNT/polymer nanocomposites are reviewed, since these properties are important, especially when these new nanocomposite materials are applied for structural applications. PMID- 16245517 TI - Clay-based polymer nanocomposites: research and commercial development. AB - This paper reviews the recent research and development of clay-based polymer nanocomposites. Clay minerals, due to their unique layered structure, rich intercalation chemistry and availability at low cost, are promising nanoparticle reinforcements for polymers to manufacture low-cost, lightweight and high performance nanocomposites. We introduce briefly the structure, properties and surface modification of clay minerals, followed by the processing and characterization techniques of polymer nanocomposites. The enhanced and novel properties of such nanocomposites are then discussed, including mechanical, thermal, barrier, electrical conductivity, biodegradability among others. In addition, their available commercial and potential applications in automotive, packaging, coating and pigment, electrical materials, and in particular biomedical fields are highlighted. Finally, the challenges for the future are discussed in terms of processing, characterization and the mechanisms governing the behaviour of these advanced materials. PMID- 16245519 TI - Synthesis of carbon nanotubes. AB - Carbon nanotubes play a fundamental role in the rapidly developing field of nanoscience and nanotechnology because of their unique properties and high potential for applications. In this article, the different synthesis methods of carbon nanotubes (both multi-walled and single-walled) are reviewed. From the industrial point of view, the chemical vapor deposition method has shown advantages over laser vaporization and electric arc discharge methods. This article also presents recent work in the controlled synthesis of carbon nanotubes with ordered architectures. Special carbon nanotube configurations, such as nanocoils, nanohorns, bamboo-shaped and carbon cylinder made up from carbon nanotubes are also discussed. PMID- 16245520 TI - Development of a resin curing model for UV nanoimprint. AB - UV nanoimprint lithography uses UV light as an energy source. It is performed at room temperature and low pressure, and has its own merits as compared to thermal nanoimprint. In this paper, a measurement system was developed to measure the degree of resin curing in UV nanoimprint to improve our understanding of the resin solidification phenomenon. A curing model was then established based on the measurement results. The measurement system measured the degree of cure in real time and was composed of a Fourier transform infrared spectroscopy system, a UV light source, and an optical guide. Also, new UV-curable resins that had low viscosity values were developed for the UV nanoimprint process, and imprint tests using these resins were performed successfully. The curing model considered the UV irradiation time, power, and curing temperature, which are important parameters in the UV nanoimprint process. The degree of cure had an exponential relation to UV irradiation time, power, and temperature; thus, the curing model was expressed as an exponential function of the UV irradiation time, power, and temperature. The developed model was verified for various UV-curable resins. PMID- 16245521 TI - Synthesis and thermal behaviour of nanostructured ZrO2 powders obtained under hydrothermal conditions. AB - Nanocrystalline zircone (ZrO2) powders were synthesized from zirconium oxyacetate aqueous solution treated up to 240 degrees C under autogeneous pressure. Depending on experimental conditions, colloidal particles of 26-150 nm are formed by aggregation of zirconia crystallites. The effect of some experimental conditions over the colloidal particle size or zirconia phase composition was studied. The powder sintering behaviour is also presented. PMID- 16245522 TI - Micro orientation and anisotropy of conductivity in liquid crystalline polymer films filled with carbon nanotubes. AB - In this communication we report the preferential orientation of single wall carbon nanotubes (SWNT) in a nematic liquid crystalline (LC) polymer matrix. The alignment of the nanotubes was characterized through anisotropy of electrical conductivity of the composite measured in directions parallel and perpendicular to the nematic director. The anisotropy of the nanocomposite films strongly depends on the nanotube concentration in the range from 1 to 10% and vanished at higher loads. The electrical conductivity of nanocomposites is related to their structural features revealed by atomic force microscopy and Raman spectroscopy experiments and is explained by a strong coupling between the nanotubes and the polymer matrix. PMID- 16245523 TI - Nanodissection of single- and double-stranded DNA by atomic force microscopy. AB - Nanodissection of single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA) has been investigated by atomic force microscopy (AFM). It is found that both ss- and dsDNA can be repeatedly dissected by an AFM tip. However, a comparison study indicates that ssDNA is a little bit more easily broken by the AFM tip than dsDNA. This is supported by the fact that the time requested to break ssDNA is shorter than that of dsDNA in the same dissection procedure under the same load. Our experiment also shows that dissection of the DNA strand is very sensitive to the load applied, and a small change of the load lead to different results. PMID- 16245524 TI - Nanomagnetic behavior of fullerene thin films in Earth magnetic field in dark and under polarization light influences. AB - In this paper magnetic fields intensity of C60 thin films of 60 nm and 100 nm thickness under the influence of polarization lights are presented. Two proton magnetometers were used for measurements. Significant change of magnetic field intensity in range from 2.5 nT to 12.3 nT is identified as a difference of dark and polarization lights of 60 nm and 100 nm thin films thickness, respectively. Specific power density of polarization light was 40 mW/cm2. Based on 200 measurement data average value of difference between magnetic intensity of C60 thin films, with 60 nm and 100 nm thickness, after influence of polarization light, were 3.9 nT and 9.9 nT respectively. PMID- 16245525 TI - Biosynthesis of gold and silver nanoparticles using Emblica Officinalis fruit extract, their phase transfer and transmetallation in an organic solution. AB - The design, synthesis and characterization of biologically synthesized nanomaterials have become an area of significant interest. In this paper, we report the extracellular synthesis of gold and silver nanoparticles using Emblica Officinalis (amla, Indian Gooseberry) fruit extract as the reducing agent to synthesize Ag and Au nanoparticles, their subsequent phase transfer to an organic solution and the transmetallation reaction of hydrophobized silver nanoparticles with hydrophobized chloroaurate ions. On treating aqueous silver sulfate and chloroauric acid solutions with Emblica Officinalis fruit extract, rapid reduction of the silver and chloroaurate ions is observed leading to the formation of highly stable silver and gold nanoparticles in solution. Transmission Electron Microscopy analysis of the silver and gold nanoparticles indicated that they ranged in size from 10 to 20 nm and 15 to 25 nm respectively. Ag and Au nanoparticles thus synthesized were then phase transferred into an organic solution using a cationic surfactant octadecylamine. Transmetallation reaction between hydrophobized silver nanoparticles and hydrophobized chloroaurate ions in chloroform resulted in the formation of gold nanoparticles. PMID- 16245526 TI - Electrical conductance in a single carbon nanofiber. AB - A microwave-plasma enhanced chemical-vapor-deposition (MPECVD) method was used to grow a solo multi-wall carbon nanofiber, which plays as a bridge across nickel electrodes that were separated by the photolithographic process. The length and diameter of carbon nanofiber are 3 microm and 100 nm, respectively. The single wire across the electrodes reveals a step current-voltage characteristic measured at high currents and low temperatures while shows a continuous behavior for multiple nanofibers. This stepwise conductance can be successfully dwelled by the quasi one-dimensional transport theory of conductors without considering the electron-phonon interaction at low temperatures and is expected to play a crucial role to determine the electrical behavior of these nanodevices. PMID- 16245527 TI - Nanoscale characterization of carbazole-indole copolymers modified carbon fiber surfaces. AB - Polycarbazole, carbazole and indole containing copolymers were electrochemically coated onto carbon fiber. The resulting polymers and copolymers were characterized by scanning electron microscopy, atomic force microscopy, X-ray photoelectron spectroscopy, and Raman spectroscopy. Characterization of the thin polymer films were performed on the polymer-coated surface of the carbon fiber. Therefore, the results obtained could elucidate the relationship between the initial feed monomer ratio, the resulting polymer/copolymer film morphology and the surface structure formed. The thickness increase (in diameter) was 0.3 and 0.9 microm, for two different composition of carbazole/indole on the carbon fiber. The carbon fibers coated with copolymer thin films were from 6.5 to 8.2 microm in diameter (from AFM measurement). PMID- 16245528 TI - Optical and field emission properties of Zinc Oxide nanostructures. AB - Zinc Oxide (ZnO) nano-pikes were produced by oxidative evaporation and condensation of Zn powders. The crystalline structure and optical properties of the ZnO nanostructures (ZnONs) greatly depend on the deposition position of the ZnONs. TEM and XRD indicated that the ZnONs close to the reactor center, ZnON-A, has better crystalline structure than the ZnONs away from the center, ZnON-B. ZnON-A showed the PL and Raman spectra characteristic of perfect ZnO crystals, whereas ZnON-B produced very strong green emission band at 500 nm in the photoluminescence (PL) spectrum and very strong Raman scattering peak at 560 cm( 1), both related to the oxygen deficiency due to insufficient oxidation of zinc vapor. ZnON-B exhibited better field emission properties with higher emission current density and lower turn-on field than ZnON-A. PMID- 16245529 TI - Uptake of silica-coated nanoparticles by HeLa cells. AB - Nanoparticles have seen wide applications in cellular research and development. One major issue that is unclear is the uptake of nanoparticles by cells. In this study, we have investigated the uptake of silica-coated nanoparticles by HeLa cells, employing rhoadime 6G isothiocyanate (RITC)-doped nanoparticles as a synchronous fluorescent signal indicator. These nanoparticles were synthesized with reverse microemulsion. A few factors, such as nanoparticle concentration, incubation time and temperature, and serum and inhibitors in culture medium were assessed on the nanoparticle's cellular uptake. The experimental results demonstrated that uptake was maximum after a 6 h incubation and was higher at 37 degrees C than that at 4 degrees C. Nanoparticle uptake depended on the nanoparticle concentration and was inhibited by hyperosmolarity, K+ depletion. In addition, serum in culture medium decreased the cellular uptake of nanoparticles. The results indicated that the uptake of silica-coated nanoparticles by HeLa cells was a concentration-, time-, and energy-dependent endocytic process. Silica coated nanoparticles could be transported into HeLa cells in part through adsorptive endocytosis and in part through fluid-phase endocytosis. PMID- 16245530 TI - Glucose biosensor based on carbon nanotube epoxy composites. AB - A novel glucose biosensor based on a rigid and renewable carbon nanotube (CNT) based biocomposite is reported. The biosensor was based on the immobilization of glucose oxidase (GOx) within the CNT epoxy-composite matrix prepared by dispersion of multi-wall CNT inside the epoxy resin. The use of CNT, as the conductive part of the composite, ensures better incorporation of enzyme into the epoxy matrix and faster electron transfer rates between the enzyme and the transducer. Experimental results show that the CNT epoxy composite biosensor (GOx CNTEC) offers an excellent sensitivity, reliable calibration profile, and stable electrochemical properties together with significantly lower detection potential (+0.55 V) than GOx-graphite epoxy composites (+0.90 V; difference deltaE = 0.35 V). The results obtained favorably compare to those of a glucose biosensor based on a graphite epoxy composite (GOx-GEC). PMID- 16245531 TI - Microstructure and magnetic properties of Fe(x)Ni(1-x) alloy nanoplatelets. AB - Plate-like nanoparticles (or nanoplatelets) of Fe(x)Ni(1-x) (x = 0.1, 0.2, 0.3, 0.4, 0.5, and 0.6) alloy were successfully synthesized through a simple sonochemical method. The shapes of the alloy nanoplatelets with different Fe atom contents are almost same. Their average diameters are about 50 nm, and their average thicknesses are several nanometers. The obtained Fe(x)Ni(1-x) alloy nanoplatelets are single-phased and have a face-centered cubic (FCC) crystal structure. The lattice constants of the alloy nanoplatelets are larger than the corresponding bulk value and increase with increasing Fe content. The surface oxidation of the alloy nanoplatelets leads to the lattice expansion. The alloy nanoplatelet powders are all ferromagnetic, and their saturation magnetizations are slightly lower than the corresponding bulk value. The saturation magnetic field and the coercivity increase with increasing Fe content. Magnetic hysteresis loops along the directions deviating different angles from the nanoplatelets plane are obviously different, indicating that the easy-axis is in the in-plane direction and the magnetization reversal is incoherent mode. The micromagnetic simulation results for the array composed of thirty-six Fe0.6Ni0.4 alloy nanoplatelets fit well with the measured data. PMID- 16245533 TI - Surface-initiated ATRP of HEA from nanocrystal alpha-Fe2O3 under ultrasonic irradiation. AB - Core/shell poly(beta-hydroethyl acrylate) (PHEA) encapsulated nanocrystal alpha Fe2O3 nanospheres (Fe2O3@PHEA) were successfully prepared by the surface initiated atom transfer radical polymerization (SI-ATRP) of beta-hydroethyl acrylate (HEA), a functional monomer, from the surfaces of the nanocrystal Fe2O3 modified with bromo-acetamide groups with the catalysts of 1,10-phenanthroline and Cu(I)Br under ultrasonic irradiation at room temperature in water. The products, Fe2O3@PHEA, were characterized by elemental analysis (EA), FT-IR, XRD, XPS and TEM. The percentage of grafting (PG%) of 38.95% and the conversion of HEA (C%) of 14.29% at room temperature for 6 h with ultrasonic irradiation were higher than the 22.41% and 8.22%, respectively, found for samples prepared by electromagnetic stirring. This demonstrated that the ultrasonic irradiation improves the SI-ATRP of HEA. PMID- 16245532 TI - Chemically functionalized water soluble single-walled carbon nanotubes modulate neurite outgrowth. AB - We report the use of chemically-functionalized water soluble single-walled carbon nanotube (SWNT) graft copolymers for modulation of outgrowth of neuronal processes. The graft copolymers were prepared by the functionalization of SWNTs with poly-m-aminobenzene sulphonic acid and polyethylene glycol. When added to the culturing medium, these functionalized water soluble SWNTs were able to increase the length of various neuronal processes. PMID- 16245534 TI - Y-junction multibranched carbon nanofibers. AB - Multibranched carbon nanofiber (CNF) is produced by a thermal chemical vapor deposition method using camphor as precursor. Nickel and cobalt catalyst was deposited on silicon substrate by e-beam evaporation and used as substrate for the growth of carbon nanomaterials. Branched carbon nanofibers were grown on the nickel thin film at 900 degrees C, whereas spherical carbon beads formed on the cobalt thin film. These fibers followed base growth mechanism devoid of any catalyst particle at the tip of fibers. PMID- 16245535 TI - Synthesis of gold nanospheres and nanotriangles by the Turkevich approach. AB - Gold nanoparticles of triangular morphology possess interesting optical properties with potential application in medicine and infrared absorbing coatings, however, little is known about conditions that favor their growth. In this paper, we have reinvestigated a time-tested recipe for the formation of gold nanospheres by citrate reduction of aqueous gold ions under boiling conditions (Turkevich recipe). Our principle findings are that gold nanotriangle formation is kinetically controlled and is highly favored at low temperatures. Furthermore, the presence of chloride ions from the precursor chloroaurate ions plays a major role in promoting the growth of <111> oriented triangular/truncated triangular particles. The presence of bromide and iodide ions that possess the ability to replace surface-bound chloride ions inhibits triangle formation to varying degrees. PMID- 16245536 TI - Effect of nanoparticle nature on hydrogen concentration profiles and improved switching characteristics in Gd switchable mirrors. AB - A detailed elastic recoil detection analysis using 40 MeV 28Si5+ ions has been carried out to study the changes in the H concentration and concentration profiles during the hydrogenation/dehydrogenation process in polycrystalline and nanoparticle Gd layers formed using vacuum evaporation and inert gas evaporation techniques, respectively. Nanoparticle sample exhibits a larger difference in the [H]/[Gd] values (2.9 and 1.7) in comparison to polycrystalline sample (2.4 and 2.0) in the loaded and deloaded states. Hydrogenation/dehydrogenation activity is restricted to the top portion in case of polycrystalline sample. In contrast to this, size induced structural transformation; enhanced surface area and the presence of large number of inter particle boundaries due to nanoparticle character result in the complete Gd layer becoming active during switching. PMID- 16245537 TI - Characterization of La2Mo2O9 aerogels synthesized by the sol-gel chemistry and high-temperature supercritical drying. AB - A lanthanum molybdate aerogel, La2Mo2O9, with a mean particle size in the range from 100 to 150 nm, was synthesized by the sol-gel method and high-temperature supercritical drying. In this communication it is shown that control over the crystallinity of product aerogels can be exercised by changing the amount of water used for hydrolysis and the temperature for subsequent heat treatment. Methoxy species are formed on the surface during synthesis. The new aerogel may prove useful as a catalyst for the oxidation of hydrocarbons to oxygenated organic compounds. PMID- 16245538 TI - Sol-gel synthesis of luminescent InP nanocrystals embedded in silica glasses. AB - III-V semiconductor nanocrystals rarely exist as spherical inclusions inside glasses, due to difficulties during their preparation, such as high toxic reagents or fast oxidation under usual glass technology temperatures. In this letter a sol-gel method for synthesis of InP nanocrystals embedded in silica glasses was described. Gels were synthesized by hydrolysis of a complex solution of Si(OC2H5)4, InCl3.4H2O, and PO(OC2H5)3. Then, the gels were heated at 600 degrees C in the presence of H2 gas to form fine cubic InP crystallites. Raman spectrum showed an InP longitudinal-optic mode (342 cm(-1)) and a transverse optic mode (303 cm(-1)). The size of InP nanocrystals was found to be from 2 to 8 nm in diameter by transmission electron microscopy. A strong photoluminescence with a peak at 856 nm was observed from InP nanocrystals embedded in silica glasses. The results suggest that it might be possible to synthesize other III-V semiconductor nanocrystals embedded in silica glasses through the sol-gel process. PMID- 16245539 TI - Supercritical fluid route for synthesizing crystalline Barium Strontium Titanate nanoparticles. AB - Pure and well-crystallized Barium Strontium Titanate (BST) nanoparticles with controlled Ba/Sr ratio have been successfully synthesized under supercritical conditions using a continuous-flow reactor in the temperature range of 150-380 degrees C at 26 MPa. To synthesize the Ba0.6Sr0.4TiO3 composition, alkoxides, ethanol and water were used. The resulting nanopowder consists of fine particles with an average particle size of 23 nm. The results show that the Ba/Sr ratio of this powder can be accurately controlled from the composition of precursor. The characterization of the as-synthesized Ba0.6Sr0.4TiO3 solid-solution and the dielectric properties of the sintered ceramics are here reported. PMID- 16245540 TI - Fabrication of a one-dimensional array of nanopores horizontally aligned on a Si substrate. AB - A one-dimensional array of nanopores horizontally aligned on a silicon substrate was successfully fabricated by anodic aluminum oxidation (AAO) using a modified two-step procedure. SEM pictures show clear nanostructures of well-aligned one dimensional nanopore arrays without cracks at the interfaces of the sandwiched structures. The processes are compatible with the planar silicon integrated circuit processing technology, promising for applications in nanoelectronics. The formation mechanism of a single nanopore array on Si substrates was also discussed. PMID- 16245541 TI - Nanoparticle delivery by controlled bacteria. AB - We present a new approach for nano-object directional delivery by bacteria based upon a taxis-controlled mechanism. In this method, a stimulus is used to direct the bacteria's motion. When carrying nano-objects, the bacteria demonstrated the ability to deliver the "loads" to targets where the stimulus is positioned. The scheme of using taxis for targeted delivery may hold a promising future for a new route to bridge nanotechnology and biotechnology. PMID- 16245542 TI - Synthesis of nanosize mesoporous MCM-48 material. AB - Nanosize mesoporous material MCM-48 was synthesized under the hydro-thermal system. The results of TEM and SEM show that the obtained MCM-48 is well organized on a 100 A length scale. Both the Nanosize and the superior organization provide this material with good thermal stability. PMID- 16245543 TI - Switches from pi- to sigma-bonding complexes controlled by gate voltages. AB - A conjugated polymer/metal ion/liquid-crystal molecular system was set between source and drain electrodes with a 100 nm gap. When gate voltage (Vg) increases, the current between source and drain electrodes increases. Infrared spectra show this system to be composed of pi and sigma complexes. At Vg = 0, the pi complex dominates the sigma complex, whereas the sigma complex becomes dominant when Vg is switched on. Calculations found that the pi complex has lower conductivity than the sigma complex. PMID- 16245544 TI - [Patent rights and the free research. The boundary between basic research and private sector is not longer razor-sharp]. PMID- 16245545 TI - [Time to break the silence! Physicians of the rich world must take responsibility for the global HIV epidemic]. PMID- 16245547 TI - [Triage--a method for the best possible care in the emergency department]. PMID- 16245546 TI - [Serotonin reuptake inhibitors may increase the risk of osteoporosis]. AB - Remodelling of the skeleton is initiated by multinucleated osteoclasts resorbing either trabecular or cortical bone. Termination of the resorptive phase is followed by new bone formation by osteoblasts. Under normal conditions, remodeling is balanced and bone mass is preserved, with the amount of new bone formed equaling the amount that was initially resorbed. Skeletal remodeling is thought to be controlled by several hormones and many different cytokines. Recent studies have indicated that signaling molecules found in the nervous system may also play an important role in bone remodeling. One of these signaling molecules is thought to be serotonin (5-hydroxytryptamin; 5-HT). Osteoblasts and osteoclasts have receptors for serotonin and both cells express the membrane protein that is responsible for serotonin reuptake (5-HTT). Gene knockout studies in mice and use of selective serotonin reuptake inhibitors (SSRls) in mice and humans have suggested that inhibition of 5-HTT leads to reduced bone mass. These observations raise the possibility that patients treated with SSRIs may have an increased risk for secondary osteoporosis and skeletal fractures. PMID- 16245548 TI - [Twenty-five years with parental groups in child health services--what is the current status?]. PMID- 16245549 TI - [New Nordic nutrition recommendations 2004. Physical activity as important as good nourishing food]. AB - The 4th edition of the Nordic Nutrition Recommendations (NNR 2004) now includes detailed recommendations on daily physical activity. For adults a minimum of 30 minutes of daily physical activity of moderate intensity and/or vigorous intensity is recommended. More physical activity (about 60 minutes daily) may be needed for prevention of weight gain. For children a minimum of 60 minutes of physical activity every day is recommended. As in the 3rd edition, a limitation of the intake of saturated plus trans fatty acids to about 10 % of the total energy intake (E%) and of the total fat intake to 30 E% is recommended. The intake of carbohydrates and dietary fibre should increase, while the intake of refined sugars should not exceed 10 E%. The 4th edition includes changes in the recommended intakes for e.g. vitamin A, D and C and for folate. PMID- 16245550 TI - [Incident analysis instead of punishment has resulted in improved patient safety in the USA]. PMID- 16245551 TI - [The Medical Biobank--a unique resource for medical research]. PMID- 16245552 TI - [A law for the management of patient data in health services. A task for the Patient Data Survey]. PMID- 16245553 TI - [We can't stay silent when people's lives and health care are jeopardized!]. PMID- 16245554 TI - [Scaremongering Cancer Fund raising propaganda is unethical]. PMID- 16245555 TI - [Integrative medicine at the Karolinska Institute--research on scientific basis]. PMID- 16245556 TI - [Fatal cases caused by fentanyl patches--remaining lethal doses in the used patches!]. PMID- 16245557 TI - [Increased PSA level in one person--myself!]. PMID- 16245558 TI - [Why was whooping cough vaccination in Sweden interrupted between 1979-1996?]. PMID- 16245559 TI - [Prostatic cancer diagnosis: waiting to start waiting]. PMID- 16245560 TI - [Being in a limbo]. PMID- 16245561 TI - [Why there is no recommendation for ultrasonography in maxillary sinusitis?]. PMID- 16245562 TI - [Brain death diagnostics and organ donation--comments to the new regulations]. PMID- 16245563 TI - [Experimental visualization of chromoneme as one of the higher levels of chromatin compactization in the mitotic chromosome]. AB - We succeeded to visualize the chromoneme or a filamentous chromatin structure, with the mean thickness 0.1-0.2 microm, as a higher level of chromatin compactization in animal and plant cells at different stages of chromosome condensation at mitotic prophase and during chromatid decondensation at telophase. Under the natural conditions, chromoneme elements are not detected in the most condensed chromatin of metaphase chromosomes on ultrathin sections. We studied the ultrastructure and behavior of the chromatin of mitotic chromosomes in situ in cultured mouse L-197 cells under the conditions selectively demonstrating the chromoeneme structure of the mitotic chromosomes in the presence of Ca2+. Loosely packaged dense chromatin bands, ca. 100 nm in diameter, chromonemes, were detected in chromosome arms in a solution containing 3 mM CaCl2. When transferred in a hypotonic solution containing 10 mM tris-HCl, these chromosome swelled, lost the chromoneme level of structure, and rapidly transformed in loose aggregates of elementary DNP fibrils, 30 nm in diameter. After this decondensation in the low ionic strength solution, the chromoneme structure of mitotic chromosomes was restored when they were transferred in a Ca2+ containing solution. The morphological characteristics of the chromoneme and pattern of its packaging in the chromosome were preserved. However, when the mitotic cells with chromosomes, in which the chromoneme structure was visualized with the help of 3 mM CaCl2, were treated with a photosensbilizer, ethidium bromide, and illuminate with a light with the wavelength 460 nm, chromatic decondensation under the hypotonic solution was not observed. The chromoneme elements in a stabilized chromatin of the mitotic chromosome preserved specific interconnection and their general pattern of packaging in in the chromatic was also preserved. The chromoneme elements in the chromosomes stabilized by light preserved their density and diameter even in a 0.6 M NaCl solution, which normally leads to chromoneme destruction. An even more rigid treatment of the stabilized chromosomes with a 2 M NaCl solution, which normally fully decondenses the chromosomes, made it possible to detect a 3D reticular skeleton devoid of any axial structures. PMID- 16245564 TI - [Determination and differentiation in the light of molecular genetics]. AB - A review of the data obtained by the author and his collaborators in studying tissue specific esterase of Drosophila males. Patterns were established for molecular-genetic regulation of synthesis of this isozyme. PMID- 16245565 TI - [Regulation of multimeric structure of NO-synthase as a new factor of organogenesis]. AB - The effect of NO on organogenesis in Drosophila is discussed. A new model of regulation of the activity of NO-producing enzyme, NO synthase is described, which takes into account endogenous synthesis of its reduced isoform. The reduced isoform of NO synthase is capable of suppressing the enzymatic activity of full sized NO synthase during formation of a heterodimer in vivo and in vitro. The reduced form of this enzyme inhibits the antiproliferative effect of the full sized NO synthase isoform during formation of eye structure in Drosophila by affecting the pathways of cell cycle regulation. The reduced form of NO synthase is an endogenous dominant-negative factor of regulation of the NO synthase activity in development of Drosophila. PMID- 16245567 TI - [Development of morphological polarity in embryogenesis of Cnidaria]. AB - A brief review dedicated to the relationship between the egg animal-vegetal axis and polarity of future larva in Cnidaria. Possible variants of changes in polarity of the embryo during development are discussed. PMID- 16245566 TI - [Problems of studying the genetic background of speciation as examplified by Drosophila virilis group]. AB - Possible ways of studying the genetic bases of speciation have been shown on the example of published data and authors' results. The data were used, which were obtained on different Drosophila species. Possible application of mapping by localization of the gene of quantitative traits and genetic transformation for solution of the problems of speciation is discussed. PMID- 16245568 TI - [Report and abstracts of the XIV Workshop on Current Problems of Developmental Biology and Biotechnology]. PMID- 16245569 TI - [Molecular basis of symbiogenic evolution: from free-living bacteria towards organelles]. AB - Molecular mechanisms of the bacteria evolution are addressed in the context of the theory of symbiogenic origin of eukaryotic cells. In the evolution of symbiotic bacteria two strategies are implemented: (a) combinative, resulted in the formation of symbiotic (sym) gene systems from the genes previously involved in the autonomous life (facultative and ecologically obligatory symbioses); (b) reductive, related to the loss of genes for the autonomous life (genetically obligatory symbioses). Both strategies are based on the increase in genomic plasticity leading: (a) during combinative evolution--to the segregation of sym gene systems into the special plasmids or "islands" (the genome size and complexity increase); (b) during the reductive evolution--to the losses of many metabolic pathways (genome size decreases). These processes are continued during the evolution of mitochondria, hydrogenosomes and plastids in which many genes for the transcription and translation were lost, while the genomes of organelles and nuclei recombined. These reorganizations are related to the peculiarities of the bacteria population dynamics within the symbiotic systems. The bacteria which combine the abilities for symbiotic and autonomous lifestyles are characterized by ecotypic polymorphism (stable coexistence of symbiotic and asymbiotic genotypes). A key role in their evolution is implemented by horizontal transfer of sym genes that in combination with different forms of natural selection (individual, frequency-dependent and group selection) are responsible for the combinative evolution of bacteria genome. In the populations of obligatory symbionts, the genetic drift and group selection dominate that ensure bacteria genome reduction, loss of their biological identity and transformation into organelles. PMID- 16245570 TI - [Animals (Animalia) in system of organisms. 2. Phylogenetic understanding of animals]. AB - The development of systematics in last decade has shown that typological classifications of five-six Kingdoms is not adequate for describing the diversity of organisms. Information from the sequences of small subunit rRNA is not sufficient to reconstruct the position of eukaryotes on the phylogenetic tree due to the effect of long branches. Totally new reconstruction of eukaryotic phylogeny was built upon the analysis of many new molecular markers. Evolution of eukaryotes had two mainstreams. One has been connected with diversification of ancestral biciliate forms (Bikonta). Sister-group of Bikonta (Unikonta) includes some originally uniciliate amoebae and moulds (Amoebozoa), and uniciliate eukaryotes with posterior cilium (Opisthokonta). The taxon Opisthokonta unites Fungi, Nuclearimorpha, Mesomycetozoa, Choanozoa and Metazoa. The latter three groups or only Metazoa are attributes to animals. The following differentiation of the groups used in systematic for the description of diversity of organisms is proposed. (1) Taxon is a group which is defined on the basis of ancestry: taxon includes all species descended from one ancestor. Taxon differs from logic classes of typology at an ontologic level. Taxon arises and exists, and its composition and occupied niches can constantly change; taxon can flourish or, on the contrary, fade up to full disappearance. Thus, the predicative characteristic of taxon, including characters which are considered significant, are not absolute. It is significant only at the moment of consideration. But characters (synapomorphies) are important as the practical tool for discerning taxa at given time period. Taxa unite species into unique classification. This understanding of taxon corresponds to monophyletic group sensu Willi Hennig. (2) Class of organisms is a group which is defined on the basis of characters: class includes all species having the given character. The class is only a logic object. Unlike taxa grouping species into classes may be through different and crossed classifications. Inside the given category of groups it is possible to distinguish: (2.1) Level of the organization (grade) described by the differences on the levels of organization: for example prokaryotic and eukaryotic levels of the organization. Eukaryotes can be divided into unicellular (Protoctista, Protista) and multicelluar (tissue-specific-Histonia) forms. (2.2) Types of the organization distinguishing groups of one level: for example, amoedoid type (Sarcodina), naked (Gymnamoebia), and testate (Testacea) amoebas. (2.3) Taxonomic groups as set-theoretical approximations of taxa. (2.4) Groups of the mixed nature. For example, Haeckel has recognized Protophyta and Protozoa describing the unicellular level of the organization inside plants and animals accordingly. Protozoa in Cavalier-Smith's system (2002, 2004) is also an example of groups of the mixed nature. PMID- 16245571 TI - Evolutionary genetics of aging in Daphnia. AB - Evolutionary theory of aging stipulates that aging is inevitable consequence of low effectiveness of natural selection acting on traits expressed late in the life span of the organisms. Two main hypotheses exist: the neutralist mutation accumulation theory and selectionist antagonistic pleiotropy theory. Both theories predict the increase of genetic variance with age; the antagonistic pleiotropy theory also predicts negative genetic correlation between fitness related traits in the beginning and in the end of the life span. In order to test these predictions we measured life expectancy and age specific mortality in cohorts of 26 c lones of Daphnia magna extracted from a single cyclic parthenogen population. Simultaneously, fecundity and age to maturity were measured in representatives of the same clones. Log mortality increased linearly with age, with little evidence for leveling off, although some replicate cohorts did show a significant leveling off of mortality. Genetic variance of log mortality was significantly higher in the last quarter of the life span than in earlier time intervals. There was a significant positive genetic correlation between early fecundity and early mortality, but not between early fecundity and late mortality. This indicates that, although there is a trade-off between fecundity and survival, this trade-off is not based on pleiotropy across ages and therefore the data does not support the prediction of antagonistic pleiotropy theory. PMID- 16245572 TI - [Calculating the intrinsic growth rate: comparison of definition and model]. AB - It was shown that well known equation r = ln[N(t2)/N(t1)]/(t2 - t1) is the definition of the average value of intrinsic growth rate of population r within any given interval of time t2-t1 and changing arbitrarity its numbers N(t). The common opinion considering the equation as suitable only for exponentially growing population was found to be incorrect. The fundamentally different approach is based on the calculation of r within the framework of demographic model, realized as Euler - Lotka equation or population projection matrices. However this model requires simultaneous realization of several assumptions improbable for natural populations: exponential change in population size, stable age structure and maintaining constant age-dependent birth and death rates. The calculation of r by definition requires the data on the dynamics of population numbers, whereas calculation on the basis of the model requires the demographic tables of birth and death rate, but not the population numbers. With the example of American ginseng it was shown that evalution of r by definition and model approaches could produce opposite results. PMID- 16245573 TI - [Levels of homoiothermy and homoioosmy and probable reasons determining them]. AB - Temperature of independently evolved homoiothermic animals (birds and animals) is maintained on the same level of 37-40 degrees C. Flying or preparing to fly insects are heated to the same temperature. This temperature is close to the lethal temperatures resulting in denaturation of proteins (Dolnik, 2003). Freshwater homoioosmy animals (annelids, mollusks, various groups of crustaceous, Cyclostomata and fishes) which independently occupied continental waters maintain internal salinity around 5-8% per hundred. The same level is retained in terrestrial and secondary marine animals (Khlebovich, 1974). In the case of decline in internal salinity below 5-8% per hundred the optical density of soluble tissue extracts increased, that could be explained by coagulation of tissue proteins. Therefore in the cases of homoiothermy and homoioosmy the values of regulated parameter are close to the parameters of protein complexes stability. It may be suggested that there are definitive advantages of governing regulated parameters exactly near boundaries of stability, out of which the process is terminated. Analogy could be found in control of reactions of nuclear synthesis and disintegration which are conducted under conditions which are close to the critical point of explosion. This statement can be regarded as a particular case of the Mopertui principle of the least operation. PMID- 16245574 TI - [On the significance of presumptions in phylogenetics (regarding the paper of Pesenko, 2005. Zhurnal Obshchei Biologii. V. 66. No 2)]. AB - Under brief consideration is the uses of presumptions, as a kind of a priori judgments, in phylogenetics in light of their critics by Pesenko (2005). It is shown that the concept of presumption is fully compatible with the hypothetic deductive argumentation scheme allowing to realize the parsimony principle in its epistemic interpretation. System of phylogenetic presumptions deserves future development in the framework of construing phylogenetics as a kind of the informal axiomatics. PMID- 16245575 TI - Building trust into the NHIN: Key legislation can ensure the privacy of personal health information. PMID- 16245576 TI - HIE takes shape in the states. PMID- 16245577 TI - Finding RHIOs. HIM professionals seek, fill roles in emerging health data networks. AB - HIM issues abound in health information exchange, but often it's up to HIM pofessionals to find the network and get in on the ground floor. PMID- 16245578 TI - IT office visits: QIOs join community health IT efforts with the 8th scope of work. AB - CMS's new scope of work sends QIOs to physician offices, hospitals, and home health agencies to provide free assistance implementing health information technology. PMID- 16245579 TI - Radical technology: your grandfather's telephone. AB - More than 100 years ago, a cutting-edge technology offered the potential to improve and better manage care. But like each innovation that was to follow, the telephone wasn't implemented in a day. PMID- 16245581 TI - The privacy and security advantages of EHRs. PMID- 16245580 TI - Deterministic, probabilistic, or fuzzy? A primer on the search algorithms that drive MPI quality. AB - MPI quality relies on the sophisticated algorithms that drive patient lookup. As health information exchange gains momentum, accurate and complete record matching will be more critical than ever. PMID- 16245582 TI - Occupational health. PMID- 16245583 TI - Public health and E-HIM. PMID- 16245584 TI - Update: guidelines for defining the legal health record for-disclosure purposes. PMID- 16245585 TI - The forms management process: keeping pace with EHR development. PMID- 16245586 TI - ICD-9-CM changes for fiscal year 2006. PMID- 16245587 TI - Reporting codes accurately. PMID- 16245588 TI - Across the continuum: multiple roles provide a wealth of information. PMID- 16245589 TI - Frequency shifts in a piezoelectric body due to a surface mass layer with consideration of the layer stiffness. AB - Shifts of resonant frequencies of a three-dimensional piezoelectric body of an arbitrary shape due to the addition of a thin layer of mass to its surface are studied. A first-order perturbation integral is obtained for the frequency shifts. The result generalizes that of a previous paper by considering the effect of the stiffness of the mass layer, in addition to its inertial effect. PMID- 16245590 TI - Theoretical, numerical, and experimental investigation on resonant vibrations of piezoceramic annular disks. AB - In this study, vibration characteristics of thin piezoceramic annular disks with stress-free boundary conditions are investigated by theoretical analysis, numerical simulation, and experimental measurement. The nonaxisymmetric, out-of plane (transverse), and axisymmetric in-plane (tangential and radial extensional) vibration modes are discussed in detail in terms of resonant frequencies, mode shapes, and electrical currents. Two optical techniques, amplitude-fluctuation electronic speckle pattern interferometry (AF-ESPI) and laser Doppler vibrometer (LDV), as well as the electrical impedance measurement are used to validate the analytical results. Both theoretical and experimental results indicate that the transverse and tangential vibration modes cannot be determined by the impedance analysis; hence, only resonant frequencies of extensional vibration modes are presented from the impedance analyzer. The LDV system is used to measure the resonant frequencies of transverse vibrations. However, both the transverse and extensional vibration modes and resonant frequencies of piezoceramic annular disks are obtained by the AF-ESPI method, and the interferometric fringes are produced instantly by a video recording system. Numerical results obtained by finite-element calculations are compared with those from theoretical analysis and experimental measurements. It is shown that the theoretical predictions of resonant frequencies and the corresponding mode shapes agree well with the experimental results. Good agreement between the predicted and measured electrical impedance also is found. The dependence of resonant frequencies and dynamic electromechanical coupling coefficients on the inner-to-outer radius ratio also is analyzed and discussed in this study. PMID- 16245591 TI - Resonant vibration investigations for piezoceramic disks and annuli by using the equivalent constant method. AB - This paper presents the equivalent constant method to investigate the transverse vibration of piezoceramic disks and annuli. By comparing the characteristic equations of resonant frequencies between isotropic and piezoceramic disks, the named equivalent Poisson's ratio v is derived, then the transverse vibration characteristic equation can be expressed as a single formulation for these two materials. To verify this method, characteristic equations of transverse vibration for piezoceramic disks and annuli with many different boundary conditions are discussed and calculated for resonant frequencies. Numerical calculations based on the finite-element method (FEM) also are performed, and the results agree rather well with the theoretical predictions. With the aid of the relations between frequency parameter and equivalent Poisson's ratio in explicit form, the other application of equivalent Poisson's ratio is the inverse evaluation of material constants. The laser Doppler vibrometer (LDV) and impedance analyzer are used to experimentally obtain the resonant frequencies of transverse and radial extensional vibrations, respectively. By the experimental results for the traction-free piezoceramic disk, the planar Poisson's ratio v(p) and planar electromechanical coupling coefficient k(p) are determined. PMID- 16245592 TI - Electrical measurement of a high-frequency, high-capacitance piezoceramic resonator with resistive electrodes. AB - In a thin and large area PZT-ceramics piezoresonator (PR) with relatively low resonance impedance, caused by high-frequency resonance and high PR capacitance, the effect of electrode resistivity and parasitic resistive and inductive elements in the measurement fixture results in significant distortion of the measured thickness-mode (longitudinal TL, shear TS) resonance response-resonance frequency shifts and characteristics deformation. This distortion may not allow the precise measurement of the PR characteristic frequencies, quality factor, and electromechanical coupling coefficient so essential to a complete PR and material characterization. A theoretical description of the "energy-trap" phenomena in a thickness-vibrating PR with resistive electrodes is presented. To interpret electrical measurements, the electromechanical model, including for completeness both the PR with resistive electrodes (as a system with distributed parameters) and the measurement fixture, is developed. The method of two contact points on the electrode provides deep sharpening and exact determination of the PR resonance. For the optimal disposition of the contact fingers, the resonance bandwidth of a real PR with resistive electrodes is even more pointed than that for the ideal PR. PMID- 16245593 TI - Micromachined acoustic wave resonator isolated from substrate. AB - This paper describes high Q, free-standing, narrow beam supported film bulk acoustic-wave resonators (FBARs) fabricated with silicon micromachining. The resonators are composed of metal/ZnO/metal/Si(x)Ny (or metal/ZnO/metal) composite layers, which are suspended by narrow Si(x)Ny/metal (or metal) beams to minimize energy leakage to the substrate. A layer of 0.5-microm thick parylene deposited and patterned over the Si(x)Ny/metal (or metal) beams is proven to enhance the sturdiness of the free-standing structure greatly. The highest Q (quality) factors we have obtained with this new structure are 1,587 and 769 at 2.7 and 5.1 GHz, respectively. This paper also describes the effect of removing the silicon nitride support layer (to form air-backed FBARs that do not use any supporting layer below or above piezoelectric the ZnO layer sandwiched by two metal layers). The electromechanical coupling constant (Kt2) is improved from 3.2% to 6.8% when a 0.9-microm thick silicon-nitride support layer is removed. PMID- 16245594 TI - Low voltage surface transverse wave oscillators for the next generation CMOS technology. AB - The design and performance of voltage controlled surface transverse wave oscillators (VCSTWO) in the lower gigahertz frequency range, operating on supply and tuning voltages in the 1.2 to 3.3 V range, and suitable for direct interfacing with the next generation CMOS circuits are presented. By applying the "boost" principle, as used in direct current (DC)-DC converters, to the design of the sustaining amplifier, the VCSTWO outputs are switched between 0 V and a positive peak value, exceeding the supply voltage Us, to provide safe CMOS circuit switching while keeping the radio frequency (RF)/DC efficiency to a maximum for low DC power consumption. The investigated 1.0 and 2.5 GHz VCSTWO are varactor tuned feedback-loop oscillators stabilized with two-port surface transverse wave (STW) resonators. Each VCSTWO has a DC-coupled, high-impedance switched output to drive the CMOS circuit directly, and an additional sinusoidal 50 ohmz high-power reference output available for other low-noise system applications. Phase noise levels in the -103 to -115 dBc/Hz range at 1 kHz carrier offset are achieved with 1.0 GHz VCSTWO at a RF/DC efficiency in the 21 to 29% range. The 2.5 GHz prototypes demonstrate phase noise levels in the -97 to -102 dBc/Hz range at 1 kHz carrier offset, and efficiencies range between 8 and 15%. PMID- 16245595 TI - Enhancements to GPS operations and clock evaluations using a "total" Hadamard deviation. AB - We describe a method based on the total deviation approach whereby we improve the confidence of the estimation of the Hadamard deviation that is used primarily in global positioning system (GPS) operations. The Hadamard-total deviation described in this paper provides a significant improvement in confidence indicated by an increase of 1.3 to 3.4 times the one degree of freedom of the plain Hadamard deviation at the longest averaging time. The new Hadamard-total deviation is slightly negatively biased with respect to the usual Hadamard deviation, and tau values are restricted to less than or equal to T/3, to be consistent with the usual Hadamard's definition. We give a method of automatically removing bias by a power-law detection scheme. We review the relationship between Kalman filter parameters and the Hadamard and Allan variances, illustrate the operational problems associated with estimating these parameters, and discuss how the Hadamard-total variance can improve management of present and future GPS satellite clocks. PMID- 16245596 TI - A digital beamformer for high-frequency annular arrays. AB - Digital transmit and receive beamformers for a 45-MHz, 7-element annular array are described. The transmit beamformer produces 0- to 80-Vpp monocycle pulses with a timing error of less than +/-125 ps. Up to four adjustable transmit focal zones can be selected. The dynamic receive beamformer uses a variable frequency sampling technique in which the frequency of analog-to-digital conversion on each channel is adjusted as the signals are received. The variable frequency clock signals required to trigger analog-to-digital conversion are obtained using a pair of high-frequency field-programmable gate arrays and a precision quartz oscillator. The gate arrays are also used to sum the digitized signals. A maximum receive beamformer timing error of less than +/-900 ps was obtained on each channel. The performance of the combined transmit and receive beamformer was tested by imaging wire phantoms. Images of CD-1 mice were also generated. The system produced images with a dynamic range of 60 dB. PMID- 16245597 TI - Focused beam control for ultrasound surgery with spherical-section phased array: sound field calculation and genetic optimization algorithm. AB - This study aims at a sound field calculation for the spherical-section phased array and an optimization algorithm for the focus patterns of phased array ultrasound surgery. An efficient field calculation formula represented as an explicit expression is derived by the strategies of projection and binomial expansion. An optimization algorithm based on genetic algorithm is constructed by the suitable fitness function and the selection strategies. The simulation results of 256-element spherical-section phased array show the capability of controlling focus accurately and effectively with the combined method made up of the explicit expression method and the genetic optimization algorithm. The simulation results of single focus, multiple foci, on-axial focus, and off-axial focus further convince the feasibility of three-dimensional (3-D) focus steering with excellent acoustic performances. A single focus with the focus dimension of 1.25 mm x 1.25 mm x 7 mm and with the intensity of 6080 W/cm2 is formed. The multiple-focus pattern can enlarge the treatment volume 22 times larger than that of single focus with a sonication. In addition, a comparison between the explicit expression approach and the point source approach testifies to the applicability of the explicit expression approach. The experiment and simulation results of 16 element array actually confirm the feasibility of the combined method. PMID- 16245598 TI - Microembolic signal characterization using adaptive chirplet expansion. AB - The adaptive chirplet expansion (ACE) is proposed to characterize high-intensity, transient signals from circulating microemboli. The nonnegative adaptive spectrogram based on the ACE gives a compact representation of the microembolic signal (MES) in joint-time, frequency domain. The mean instantaneous power (MIP) and mean instantaneous frequency (MIF) of MES are estimated from the adaptive spectrogram. Then, several important characteristics of MES, such as embolus-to blood ratio (EBR), half width maximum (HWM), and embolic signal onset (ESO), are computed from the MIP, and the frequency modulation is examined in the MIF. To validate the new method, we improved the simulation model of the audio Doppler ultrasound signal. Some MESs together with a Doppler ultrasound signal from carotid blood flow are simulated in the simulation study. As a comparison, the adaptive Gabor expansion (AGE) also is implemented on these simulated signals. The experimental results of the simulation study show that the new method, based on the ACE, outperforms the AGE-based method in MES characterization. The consistent conclusion has been confirmed by the clinical study on some clinical MESs. PMID- 16245599 TI - Evaluation of an ultrasonic echo-tracking method for measurements of arterial wall movements in two dimensions. AB - The longitudinal movement of blood vessel walls has so far gained little or no attention, as it has been presumed that these movements are of a negligible magnitude. However, modern high-resolution ultrasound scanners can demonstrate that the inner layers of the arterial wall exhibit considerable movements in the longitudinal direction. This paper evaluates a new, noninvasive, echo-tracking technique, which simultaneously can track both the radial and the longitudinal movements of the arterial wall with high resolution in vivo. Initially, the method is evaluated in vitro using a specially designed ultrasound phantom, which is attached to and moved by an X-Y system, the movement of which was compared with two high-resolution triangulation lasers. The results show an inaccuracy of 2.5% full scale deflection (fsd), reproducibility of 12 microm and a resolution of 5 microm, which should be more than sufficient for in vivo studies. The ability of the method is also demonstrated in a limited in vivo study in which a preselected part of the inner vessel wall of the right common carotid artery of a healthy volunteer is tracked in two dimensions over many cardiac cycles. The results show well reproducible x-y movement loops in which the recorded radial and longitudinal movements both are of the magnitude millimetre. PMID- 16245600 TI - Motion artifact reduction for IVUS-based thermal strain imaging. AB - Thermal strain imaging (TSI) using intravascular ultrasound (IVUS) has the potential to identify lipid pools within rupture-prone arterial plaques and serve as a valuable supplement to current IVUS systems in diagnosing acute coronary syndromes. The major challenge for in vivo application of TSI will be cardiac motion, including bulk motion and tissue deformation. Simulations based on an artery model, including a lipid-filled plaque, demonstrate that effective bulk motion compensation can be achieved within a certain motion range using spatial interpolation. We also propose a practical imaging scheme to minimize mechanical strains caused by tissue deformation based on a linear least squares fitting strategy. This scheme was tested on clinical data by artificially superimposing thermal displacements corresponding to different temperature rises. Results suggest a 1-2 degrees C temperature rise is required to detect lipids in an atherosclerotic plaque in vivo. PMID- 16245601 TI - A new imaging strategy using wideband transient response of ultrasound contrast agents. AB - High-resolution clinical systems operating near 15 MHz are becoming more available; however, they lack sensitive harmonic imaging modes for ultrasound contrast agent (UCA) detection, primarily due to limited bandwidth. When an UCA is driven to nonlinear oscillation, a very wideband acoustic transient response is produced that extends beyond 15 MHz. We propose a novel strategy using two separate transducers at widely separated frequencies and arranged confocally to simultaneously excite and receive acoustic transients from UCAs. Experiments were performed to demonstrate that this new mode shows similar resolution, higher echo amplitudes, and greatly reduced attenuation compared to transmission at a higher frequency, and superior resolution compared to transmission and reception at a lower frequency. The proposed method is shown to resolve two 200 microm tubes with centers separated by 400 microm. Strong acoustic transients were detected for rarefaction-first 1-cycle pulses with peak-negative pressures above 300 kPa. The results of this work may lead to uses in flow and/or targeted imaging in applications requiring very high sensitivity to contrast agents. PMID- 16245602 TI - A system for simultaneously measuring contact force, ultrasound, and position information for use in force-based correction of freehand scanning. AB - During freehand ultrasound imaging, the sonographer places the ultrasound probe on the patient's skin. This paper describes a system that simultaneously records the position of the probe, the contact force between the probe and skin, and the ultrasound image. The system consists of an ultrasound machine, a probe, a force sensor, an optical localizer, and a host computer. Two new calibration methods are demonstrated: a temporal calibration to determine the time delay between force and position measurements, and a gravitational calibration to remove the effect of gravity on the recorded force. Measurements made with the system showed good agreement with those obtained from a standard materials testing machine. The system's uses include three-dimensional (3-D) ultrasound imaging, force-based deformation correction of ultrasound images, and indentation testing. PMID- 16245603 TI - A traveling-wave, modified ring linear piezoelectric microactuator with enclosed piezoelectric elements--the "scream" actuator. AB - A 1.8 cc silent bidirectional traveling-wave, self-moving linear microactuator is shown to be capable of generating a sliding velocity of 0.22 m/s and sliding force of 1.1 N. Through the use of computational analysis in the actuator's design, the vibration characteristics were improved in order to obtain a better actuator. The generation of a radial traveling wave about the circumference of the actuator, akin to a ring, is shown to exist despite the unusual shape, and the presence of traveling wave motion along the output face also is shown to exist. By using short-time sinusoidal signals, slider displacements as small as 82 nm were obtained from the actuator, and by using direct current (DC) input, displacements of up to +/-107 nm were obtained, suggesting a way to obtain subnanometer positioning accuracy over arbitrary sliding distances. By reversing the phase between the paired driving signals, the direction of motion was reversed at up to 300 Hz; the slider displacement and velocity was found to be inversely proportional to the phase-reversal rate, and the slider's peak velocity and maximum thrust force were directly proportional to the phase between the driving signals. The output force and velocity of the actuator was fairly insensitive to the input frequency, giving measurable motion between 132.5 and 141.5 kHz, but was sensitive to the input voltage, requiring at least 38 V input for operation, and was approximately quadratically dependent on the applied preload centered about 2.25 N. PMID- 16245604 TI - Interferometric detection in picosecond ultrasonics for nondestructive testing of submicrometric opaque multilayered samples: TiN/AlCu/TiN/Ti/Si. AB - An experimental investigation of nanometric thin films by a picosecond ultrasonic technique is presented. A photoelastic model is used with an interferometric device, combined with ultrafast optical pump and probe setup, to measure the thicknesses of submicrometric layers made of TiN, Ti, and AlCu deposited on silicon (Si) wafers. The results are in good agreement with ellipsometry measurements showing that the picosecond ultrasonic technique can give accurate results even when the reflectance signal is very low. Additional important results are first, that the adhesion of the TiN surface film is probed by processing both the frequency and the damping of the oscillation of a resonance acoustic mode; and second, the presence of a thin buried TiN layer under an opaque AlCu film is highlighted by the interferometric setup. PMID- 16245605 TI - Design of ultrasonic transmitters with defined frequency characteristics for wireless pressure sensing in injection molding. AB - This paper describes a new mechanical wireless data transmission technique using ultrasonic waves as the information carrier for on-line injection mold cavity pressure measurement. Ultrasonic transmitters with specific frequency characteristics were designed, modeled, simulated, and prototyped for pressure data retrieval from an enclosed machine environment, as well as for sensor identification in a sensor matrix configuration. The effects of the front layer and bonding layer of the transmitter on the overall sensor frequency characteristics were investigated, using an equivalent circuit model. The optimal layer thickness was determined for the design of transmitters with specific dominant resonant frequency and narrow bandwidth. Experimental results were in good agreement with the analysis, thus confirming the design approach. PMID- 16245606 TI - A diffraction-based optical method for the detection of in-plane motion of lamb waves. AB - This paper describes a laser optical technique that allows the detection of in plane motion of Lamb waves. This interference-based laser optical technique includes a tiny square indentation with a width of about 30 micron on the sample surface and a relatively simple optical arrangement. The current technique is applied for the detection of in-plane motions of Lamb waves propagating in a 70 micron thick brass plate. Measurement of So mode dominated by in-plane motion in the low fd (frequency times thickness) regime is successfully demonstrated with the current technique. With the indentation replaced by a microreflector in a microelectromechanical (MEMS) structure, this technique is applicable for the detection of in-plane motion in MEMS structures. PMID- 16245607 TI - Prediction of the thermal sensitivity of surface acoustic waves excited under a periodic grating of electrodes. AB - The prediction of the temperature sensitivity of surface acoustic wave (SAW) devices still requires improvement because the nature of the implemented surface modes and the devices' complexity strongly change from the early basic Rayleigh wave-based devices. To address this problem, a theoretical analysis and a numerical tool have been developed to predict the thermal dispersion of general electro-acoustic devices. The proposed model accounts for the electrode contribution to the frequency-temperature law. The computed thermal sensitivities are compared to experimental results for different kinds of substrates and waves. PMID- 16245608 TI - Analysis and design of focused interdigital transducers. AB - Focused interdigital transducers (FIDTs) can generate surface acoustic wave (SAW) with high intensity and high beamwidth compression ratio. Owing to these features, they are very suitable to be used as the sources of microacoustic channels or waveguides in the near future. The focusing properties of FIDTs are dominated solely by their geometric shapes. Therefore, to obtain optimal performance, it is essential to analyze the FIDTs with a variety of geometric shapes. However, among the existing studies concerning the diffraction of FIDTs, a detailed analysis and design of FIDTs is still in paucity. In this paper, we adopted the exact angular spectrum of plane wave theory (ASoW) to calculate the amplitude fields of FIDTs on Y-Z lithium niobate (LiNbO3) with the shape as a concentric circular arc and the concentric wave surface. Based on the calculation results, we discussed the variations of the amplitude fields induced by changing number of pairs, degree of arc, and geometric focal length. In addition, the focusing properties of FIDTs on the (100)-oriented GaAs substrate were also analyzed and discussed. We also summarized the guiderules for designing a FIDT via four important factors. It is worth noting that the results of this study provide an important basis for designing various FIDTs to fit the desired applications. PMID- 16245609 TI - Free vibration analysis of the piezoceramic bimorph with theoretical and experimental investigation. AB - This paper investigates the vibration characteristics of an asymmetric, three layered piezoceramic circular bimorph under traction-free boundary conditions by applying the electroelasticity and Kirchhoff plate theory. The asymmetric, three layered bimorph consists of an isotropic shim layer and two piezoceramic layers of equal thickness and same polarization. Two optical techniques, amplitude fluctuation electronic speckle pattern interferometry (AF-ESPI) and laser Doppler vibrometer (LDV), are used to validate the theoretical analysis. The resonant frequencies of the piezoceramic bimorph also are measured by an impedance analyzer. Both theoretical and experimental results indicate that the transverse vibration modes cannot be measured by impedance analysis, and only resonant frequencies of extensional vibration modes are present. However, transverse vibration modes of the piezoceramic bimorph can be obtained by the AF-ESPI and LDV measurements. The numerical calculations also are obtained using the finite element method (FEM), and the results agree comparatively well with the theoretical analysis and experimental measurements. According to the theoretical calculation, the variations in resonant frequencies and effective coupling factors versus the various layer-thickness ratios also are investigated in this work. PMID- 16245610 TI - Optical piezoelectric transducer for nano-ultrasonics. AB - Piezoelectric semiconductor strained layers can be treated as piezoelectric transducers to generate nanometer-wavelength and THz-frequency acoustic waves. The mechanism of nano-acoustic wave (NAW) generation in strained piezoelectric layers, induced by femtosecond optical pulses, can be modeled by a macroscopic elastic continuum theory. The optical absorption change of the strained layers modulated by NAW through quantum-confined Franz-Keldysh (QCFK) effects allows optical detection of the propagating NAW. Based on these piezoelectric-based optical principles, we have designed an optical piezoelectric transducer (OPT) to generate NAW. The optically generated NAW is then applied to one-dimensional (1 D) ultrasonic scan for thickness measurement, which is the first step toward multidimensional nano-ultrasonic imaging. By launching a NAW pulse and resolving the returned acoustic echo signal with femtosecond optical pulses, the thickness of the studied layer can be measured with <1 nm resolution. This nano-structured OPT technique will provide the key toward the realization of nano-ultrasonics, which is analogous to the typical ultrasonic techniques but in a nanometer scale. PMID- 16245611 TI - Acoustic transmission losses and field alterations due to human scalp hair. AB - Hair is a potential transmission barrier for diagnostic and therapeutic ultrasound. We tested hair phantoms for insertion losses and field effects at 0.27-2.8 MHz. The negligible losses observed below 0.5 MHz suggest that, at such frequencies, hair removal is unnecessary for low-power imaging or therapeutic applications. PMID- 16245612 TI - Simplified unipolar, quasisquare wave energy recovery drive circuits for piezoelectric actuators. AB - A simple two-switch circuit for driving piezoelectric actuators with unipolar quasisquare waves is presented. The circuit provides for recovery of the energy stored on the actuator capacitance back to the primary power supply when the actuator is de-energized. PMID- 16245613 TI - An iterative approach to design a SAW filter having desired response. AB - Present work proposes an iterative technique to estimate reflection weighting function and transduction weighting function simultaneously to achieve a desired single-phase unidirectional transducer (SPUDT) response. The technique uses the p matrix formulation in order to describe characteristics of a SPUDT. Simulation results are presented to show the validity of the approach. PMID- 16245614 TI - What it takes: health care responds to Hurricane Katrina. Interview by Denise Dungey Reed. PMID- 16245615 TI - Consumer-directed health plans a new force in health insurance? PMID- 16245616 TI - Transaction services: beyond basics to bolt-ons. PMID- 16245617 TI - Possibilities and pitfalls of outsourcing. AB - Outsourcing can save healthcare organizations costs related to staffing and training. Organizations should ensure that a vendor's staff is credentialed, knowledgeable, and properly trained. Outsourcing firms should ensure the confidentiality and security of the information they will handle. Outsourcing carries risks for providers, including potentially negative impact on tax-exempt status and loss of control over business processes. PMID- 16245618 TI - Time for patient payment strategies. AB - A patient payment strategy should have four objectives: Establish equitable and consistent policies and communicate them clearly to patients. Facilitate appropriate payment, regardless of the payer. Increase patients' awareness of their payment options and responsibilities. Enhance patient relations. PMID- 16245619 TI - Time for a price check? AB - General acute care hospitals often are losing out to niche providers. By focusing commercial payment on proprietary and commodity services and away from noncore services, these hospitals can help guard against the competitive threat. Transitioning to the price shifting model requires distinct and careful actions. PMID- 16245620 TI - Cost-based pricing and the underperforming physician group. AB - Cost-based pricing may help healthcare organizations improve the revenue performance of their employed physician groups. Organizations considering a change to cost-based pricing strategies should be prepared to supply their major payers with the following supporting information: Physician compensation rates. Compensation methodology. New practice development. Operating benchmarks. Overhead benchmarks. PMID- 16245622 TI - Asset management: the big picture. AB - To develop an comprehensive asset management plan, you need, first of all, to understand the asset management continuum. A key preliminary step is to thoroughly assess the existing equipment base. A critical objective is to ensure that there are open lines of communication among the teams charged with managing the plan's various phases. PMID- 16245621 TI - Assessing the feasibility of developing centers of excellence: six initial steps. AB - Using these six steps, hospital administrators can begin to assess the feasibility of creating a center of excellence in their facility: Forecasting demographic changes in the service area. Identifying service lines. Calculating use rates for key service lines. Evaluating market trends and market share. Conducting a financial review. Formulating a plan. PMID- 16245623 TI - Benchmarking supply expenses: the devil's in the definition. AB - For benchmarking hospital supply expense to be meaningful, the healthcare industry needs an accurate and generally agreed-upon definition of "supply expense." PMID- 16245624 TI - Making hospital-physician collaboration work. AB - Hospital-physician collaboration can reduce costs, alleviate unproductive competition, increase revenue, and foster productive working relationships. Using structured dialogue can improve communications. A hospital medical advisory panel of physicians can champion collaboration. A three-stage framework of proactivity, collaborative conflict, and containment can help resolve physician-hospital competition. PMID- 16245626 TI - Reducing leader variance. PMID- 16245625 TI - Improving cash flow with better charge capture and denial management. PMID- 16245627 TI - How do we engage the medical staff in IT? PMID- 16245628 TI - When should you consider acquisition? PMID- 16245629 TI - Distribution of keratins and involucrin in human fetal oral epithelia. AB - The presence of keratins and involucrin was studied in 9 human fetuses of an age range between 9 and 24 weeks of gestation. The use of the monoclonal antibodies AE1 and AE3 as well as an anti-involucrin serum enabled developmental changes in the oral epithelia to be assessed. Our study detected some evidence of fetal differentiation patterns different from the adult, i.e. presence of AE3-Positive basal cells and AE1-Positive suprabasal cells in early stages of fetal development. In addition, involucrin was detected as early as 9 weeks of gestation in the superficial cells of the oral cavity epithelium. After the 15th week of gestation the differentiation markers studied gradually adopted adult type distribution patterns. PMID- 16245630 TI - Verrucous carcinoma, hyperplasia and leukoplakia of the oral mucosa: a clinico histopathological and histometric study. AB - Thirteen cases of verrucous leucoplakia (VL), 3 of verrucous hyperplasia (VH) and 19 of verrucous carcinoma (VC) were evaluated by means of 4 clinical features, 12 microscopic parameters and 6 epithelial histometric measurements. No significant clinical differences were detected, but histologic data showed that orthokeratinization was more frequent in VL and VH, while parakeratinization proved more common in VC. Sharp epithelial projections predominated in all three lesion types, though lymphoplasmatic infiltration and Russell bodies were more frequent in VH. Histometrically, there were statistical differences between VL or VH vs VC in three parameters, namely connective tissue-epithelial interface (Ice), epithelial height (He) and connective tissue-epithelial interface plus verrucous epithelial surface (Ice + Sve). To conclude, in this series, VH failed to exhibit significant clinical or histologic differences vs VL or VC, but histometric analysis was able to detect epithelial differences between both premalignant lesions and VC. PMID- 16245631 TI - Changes in the vascular bed of submandibular glands after chronic autonomic denervation. AB - The effect of chronic sympathetic denervation (SCG) or parasympathetic decentralization (PSD) on the vascularization of the submaxillary gland (SM) of rat was studied in connection with morphological and functional changes. The resection of the superior cervical ganglion or the section of the chorda tympani was performed in adult male Wistar rats prior to the injection of Microfil MV Orange 3.25 cm3/kg. Once dissected, the glands were made transparent in increasing concentrations of methyl salicylate--alcohol. Three longitudinal sections were performed and the central one was projected in an automatic image analyzer (Kontron Zeiss), where vascular length and vascular volume were measured. Another group of animals was injected with Microfil after having received an injection of noradrenaline (10 g/kg) in SCG glands and methacholine in PSD glands. In SCG glands, vascular length underwent a 60% increase and the vascular volume increased from 0.26 +/- 0.15 cu mm/cu mm in controls to 0.54 +/- 0.15 cu mm/cu mm in SCG. The intravenous administration of noradrenaline prior to Microfil injection reverted the phenomenon. PSD caused a reduction of the vascular bed which resulted mainly in increased vascular volume. The administration of methacholine (10 g/kg) reverted the-response. It was concluded that SCG induced a loss of the existing vasoconstrictor tone, thus causing increase of vascular length and volume (passive vascular dilatation). Increased vascular volume after chronic PSD is compatible with vascular stasis and the absence of variations in vascular volume after the injection of methacholine would indicate that in the SM glands of rats there is no vascular dilatation tone. PMID- 16245632 TI - Determination of heparin in clinically normal dog gingiva. AB - The purpose of this study was to develop a technique to allow the extraction, identification and quantification of heparin in small samples (0.5 g) of normal gingival tissue of the Beagle dog. Heparin was extracted following homogenization, defattening, proteolytic digestion and precipitation with organic solvents and identified by a microelectrophoretic method carried out on an agarose slide. The concentration of heparin was determined by measuring the optical density of the metachromatic toluidine blue stained spots and comparing them with standard reference heparin spots. Gingiva contained 25 +/- 1.9 units of heparin per gram of wet tissue with a relative migration rate of 0.92 +/- 0.005 vs the unit value for reference heparin. Heparin was further characterized by enzyme degradation with adapted F. heparinum (heparinase). Its anticoagulant activity was demonstrated by extending Plasma Thrombin Time, which was normalized with the anti-heparin substance protamine sulfate. PMID- 16245633 TI - Microphotometric quantitation of succinic dehydrogenase in whole isolated osteoclasts. AB - The microphotometric quantitation of succinic dehydrogenase (SDH) in whole isolated osteoclasts was performed. Osteoclasts were obtained from imprints of the external metaphyseal area of tibia, a zone of physiological bone resorption. SDH was demonstrated histochemically and the reaction product was measured in a Zeiss Cytoscan microphotometer in arbitrary units (AU) of activity from 0 to 500. Total enzyme activity (TEA) was 10 to 50 AU in 78% of the cells. The enzyme concentration (MEC), taken as the activity per unit volume was 0.10 to 0.40 AU in 94% of the cells. Values of reactive cytoplasm area (RCA) were high for most of the cells, but lower for a few, probably mononuclear, osteoclasts. The correlation index for TEA vs RCA was significant. The study of enzyme activity in whole osteoclasts by microphotometric quantitation has been proven possible. This technique could be employed in further studies of enzyme activity of osteoclasts in other physiological or pathological states of bone resorption. PMID- 16245634 TI - [CME-ECG 8. Palpitations after fall from bicycle]. PMID- 16245635 TI - [What is your diagnosis? "Tooth" in the left ovary, teratoma]. PMID- 16245636 TI - [Nanotechnology in medicine]. AB - In the past, micro and nano technology were expected to have a considerable meaning for industry and especially for life sciences. The media were prodigal with visions of a tremendous rise of the life span and an enormous amelioration of life quality due to micro and nano submarines in the body and due to novel implants. In spite of these, developments in these disciplines proved to be by far more difficult and therefore lengthy in practice than expected. It is therefore necessary to verify realistically how such high technologies may support medicine. In this article, realised works are treated and new research works presented. Developments in endoscopy, novel materials und the lab on chip reached a sophisticated state of development and are therefore to be included in clinical practice for the future. PMID- 16245637 TI - [Crohn's disease--standards of treatment 2004]. AB - In Crohn's disease therapeutic concepts are according to distinct conditions. Course of the disease, the individual disease pattern and the aim of treatment are of particular significance. Care of patients with Crohn's disease requires interdisciplinary cooperation between gastroenterologists and surgeons. Primary therapy in mild to moderate disease comprises aminosalicylates and budesonide. Treatment of refractory or severe cases are corticosteroids. Immunosuppressive therapy is indicated in all kinds of complicated disease. First line immunosuppressants are Azathioprine and 6-Mercaptopurine while Methotrexate, Infliximab, Mycophenolatmofetil and other compounds represent alternative or rescue medications. Maintenance of remission should not be done on a regular basis but rather regarding the individual patients' situation. Risks have to be carefully balanced with possible benefits. The most important aim of treatment is quality of life. PMID- 16245638 TI - [Ulcerative colitis? Guidelines 2004]. AB - Ulcerative colitis was first described in 1859 from Samuel Wilks, a physician at Guy's hospital in London. The prevalence in the high incidence areas ranges from 80 to 120/100.000/year. Ulcerative colitis is a chronic relapsing or chronic active disease which starts at the rectum and presents with a continuous inflammation. Primarily young adults are affected (20 to 40 years of age) but the disease may present at all ages, from younger than 1 year of life to the 80s. Many series show a secondary peak in incidence in the elderly. In the present review we will focus on the basic principles of the therapy with regard to the variety of disease manifestations. The therapeutic algorithms will be described separately for the induction of remission and the maintenance of remission. The localization of inflammation and disease activity represent crucial factors which have to be considered. With regard to these factors, the therapeutic regimens range from simple local therapy with aminosalicylates to systemic immunosuppressive therapy, which will in extreme cases require the administration of ciclosporin. Since ulcerative colitis is associated with an increased risk in developing colon carcinoma, medical therapy as well as endoscopic surveillance are fundamental in the prevention of carcinoma. In the end an outlook to future therapeutic targets and strategies will be provided. PMID- 16245639 TI - [CME-ultrasonography 7/solution. Chlamydia peritonitis]. PMID- 16245640 TI - Three-dimensional finite element analysis of stress in the periodontium. AB - The aim of this study is to calculate the stress produced in the periodontium at different bone levels under occlusal load. Four finite element models of maxillary incisors were designed consisting of the tooth, pulp, periodontal ligament and alveolar bone at various levels of bone height (25%, 50% and 75%). An occlusal load of 24 kg at an angle of 50 degrees to the long axis of the tooth was applied on the palatal surface, at the level of the middle third of the crown. All the models were assumed to be isotropic, linear and elastic and the analysis was performed on a Pentium IV computer using the NISA II Display III software. The results showed that maximum stress in the tooth was seen at the cervical region and to a greater extent at the apex for all the models. In periodontal ligament, maximum stress was seen at the alveolar crest in models with normal alveolar bone height and with reducing alveolar bone height maximum stress was at the apex. There was no significant stress distribution in alveolar bone, while no stress was seen on the pulp. The results of the study demonstrated a significant increase in stress concentration at the apex with loss of alveolar bone height. PMID- 16245641 TI - Peripheral giant cell granuloma combined with facial hemangioma. A case report. AB - Peripheral giant cell granuloma (PGCG) is a relatively uncommon benign tumor of the oral cavity affecting people of all ages and with no racial predilection. The etiology is still unclear but local trauma or chronic irritants may cause the development of this lesion. Hemangioma is a benign process in which there are an increased number of normal or abnormal-appearing blood vessels. In this article the clinical, radiographic and histopathologic characteristics of a patient who had a rare combination of PGCG and facial hemangioma is discussed. Surgery was the effective treatment modality for the lesion in the mandible. The characteristics of facial hemangioma and PGCG have similarities, such as the possibility of bleeding problems during surgery. As the lesion in this case was not small in size, it was not a conservative procedure. Bleeding control was established by electrocauterization. Healing was uneventful and there has been no recurrence since the surgery. A new prosthesis was fabricated for the patient. In these kinds of cases the control of bleeding must be very carefully determined and attention should be paid to the possibility of serious bleeding problems. Based on the data presented we were not able to find any correlation between the PGCG and the facial hemangioma, but new reports might have different views on the subject in the future. PMID- 16245642 TI - Treatment of human class II furcation defects using connective tissue grafts, bioabsorbable membrane, and resorbable hydroxylapatite: a comparative study. AB - OBJECTIVE: To evaluate clinical outcomes and assess digital subtraction radiographic changes after using bioabsorbable membrane (polyglycolic acid/polylactic acid--PGA/PLA) or a connective tissue graft (CTG) as a barrier, with or without resorbable hydroxylapatite (HA Resorb) in the treatment of mandibular class II furcations. METHODS: Fifty furcations in twenty patients with chronic periodontitis were divided into five treatment groups: (I) PGA/PLA; (II) PGA/PLA and resorbable hydroxylapatite; (III) CTG; IV) CTG and resorbable hydroxylapatite; and (V) flap debridement alone (control). Vertical and horizontal probing depths, vertical probing attachment level, gingival recession and standardized periapical radiographs were obtained at baseline and at 3, 6 and 12 months post-operatively. At 12 months, the treated teeth were exposed by re entry to verify clinical results. RESULTS: All experimental groups (I, II, III and IV) showed statistically significant improvement in the clinical parameters and bone density as compared to the control group. However, no statistically significant differences were observed among any of the experimental groups. Percentages of complete furcation closure and sites still defined as class II furcations were, respectively, as follows: 40% and 20-30% for groups II and IV; 20% and 40% for groups I and III; 0% and 80% for control. Groups II and IV showed significant radiographic changes in bone gain at both 6 and 12 months, while groups I and III showed significant changes only at 12 months. CONCLUSIONS: (1) Guided tissue regeneration (GTR) therapy with or without resorbable hydroxylapatite showed significantly favorable results clinically and radiographically compared to flap debridement alone. (2) GTR plus resorbable hydroxylapatite (groups II and IV) showed higher percentages of complete furcation closure and more bone gain than GTR alone (groups I and III). (3) CTG may be used as an alternative to PGA/PLA bioabsorbable membrane with comparable potential. (4) Digital subtraction radiography may give higher accuracy in assessing results of periodontal therapy. PMID- 16245643 TI - Immunohistochemical localization of cytokeratin 19, involucrin and proliferating cell nuclear antigen (PCNA) in cultured human gingival epithelial sheets. AB - It has been suggested that human cultured gingival epithelial sheets may serve as a possible grafting material. The purpose of this study was to examine the biological characteristics of human cultured gingival epithelial sheets by epithelial differentiation and proliferation markers. Immunohistochemical localization of cytokeratin 19, involucrin and proliferating cell nuclear antigen (PCNA) were examined in human cultured gingival epithelial sheets samples from twenty patients. Cytokeratin 19-immunopositive cells were scattered mainly in the suprabasal layer. Immunoreactivity for involucrin was observed in all layers except for the basal layer. The majority of proliferating cell nuclear antigen immunopositive cells was found in the basal layer. These results suggested that the cultured human gingival epithelial sheets were biologically active and in proliferative condition, which implies that this biological product may be a potential grafting material. PMID- 16245644 TI - A survey of antiviral drugs for bioweapons. AB - Smallpox (variola major), and the haemorrhagic fever viruses (filoviruses and arenaviruses) are classified as Category A biowarfare agents by the Centers for Disease Control. Category A agents pose a significant risk to public health and national security because they can be easily disseminated by aerosol, although with the exception of variola, they are not easily transmitted from person to person. An attack with these viruses would result in high morbidity and mortality and cause widespread panic. With the exception of smallpox and Argentine haemorrhagic fever virus, there are no vaccines or approved treatments to protect against these diseases. In this review we focus on promising prophylactic, therapeutic and disease modulating drugs (see Figure 1 for select chemical structures). PMID- 16245645 TI - In vitro activity of SPD754, a new deoxycytidine nucleoside reverse transcriptase inhibitor (NRTI), against 215 HIV-1 isolates resistant to other NRTIs. AB - SPD754 (also known as AVX-754) is a deoxycytidine analogue nucleoside reverse transcriptase inhibitor (NRTI) with antiretroviral activity against HIV-1 and HIV 2 in vitro and against recombinant viruses containing thymidine analogue mutations (TAMs). In order to better establish the activity of SPD754 against HIV 1 containing TAMs, twelve panels of up to twenty clinical isolates with defined TAM combinations were selected from the ViroLogic database. Phenotypic viral susceptibility to SPD754 and five other NRTIs was tested using the PhenoSense HIV assay and expressed as median fold-change compared with a reference strain. In total, 215 isolates were selected, representing four TAM patterns in both pathways by which TAMs accumulate clinically. The presence of five TAMs in the 41, 215 pathway, at codons 41, 67, 210, 215, and 219 of reverse transcriptase (RT), produced a median 1.8-fold reduction in SPD754 susceptibility, compared with fold reductions to zidovudine, lamivudine, abacavir, didanosine and tenofovir of 438, 4.8, 4.5, 1.4 and 3.6, respectively. Five TAMs in the 67, 70, 219 pathway (at codons 41, 67, 70, 215 and 219) reduced SPD754 susceptibility by a median 1.3-fold, compared with fold reductions for the aforementioned NRTIs of 108, 3.2, 3.0, 1.3 and 2.5, respectively. M184V addition reduced SPD754 susceptibility by 1.8-fold in the presence or absence of TAMs. SPD754 retains a substantial proportion of its antiviral activity against HIV-1 containing multiple TAMs, with or without the M184V mutation. These data suggest that SPD754 is a promising new NRTI for the treatment of NRTI-experienced HIV-infected patients. PMID- 16245646 TI - Novel phorbol esters exert dichotomous effects on inhibition of HIV-1 infection and activation of latent HIV-1 expression. AB - Two new phorbol esters, NPB-11 (12-O-methoxymethylphorbol-13-decanoate) and NPB 15 (12-O-benzyloxymethylphorbol-13-decanoate) were synthesized. The compounds exhibited potent anti-HIV-1 activity and low cytotoxicity in MT-4 cells by MTT assay even at a high concentration [50% cytotoxic concentrations (CC50) were 8.32 and 4.39 microg/ml, respectively]. Two inhibitors strongly suppressed HIV-1 (IIIB strain) replication in MT-4 cells with a 50% effective concentration (EC50) of 1.3 and 0.27 ng/ml, respectively. NPB-11 efficiently blocked replication of both X4 and R5 HIV-1 in PHA-activated peripheral blood mononuclear cells and MT-4 cells as revealed by p24 assay. The antiviral activity appeared to be mediated, at least partially, by the down-regulation of the expression of CD4 and the HIV-1 co-receptors, CXCR4 and CCR5. The compounds were also capable of selectively up regulating HIV-1 expression in a variety of latently infected cell lines and inducing cell death in HIV-1 infected cells. The effect of NPBs on the induction of HIV-1 was specifically blocked by nontoxic doses of a protein kinase C blocker, staurosporine. NPB-11 blocked the spread of HIV-1 released from latently infected ACH-2 cells to MT-4 cells in a co-culture system. When combined with AZT, NPB-11 synergistically inhibited HIV-1 replication in MTT assay using MT-4 cells. These data suggest that these agents might be useful in reducing persistent viral reservoirs in patients and as adjuvant therapy in patients treated with HAART. PMID- 16245647 TI - Synthesis and biological activity of 1H-benzotriazole and 1H-benzimidazole analogues--inhibitors of the NTpase/helicase of HCV and of some related Flaviviridae. AB - To improve anti-helical activity of analogues of 1H-benzotriazole and 1H benzimidazole their N-alkyl derivatives were synthesized and tested for antihelicase activity against enzymes of selected Flaviviridae including hepatitis C virus (HCV), West Nile virus (WNV), Dengue virus (DENV) and Japanese encephalitis virus (JEV). 1- and 2-alkyl derivatives of 4,5,6,7-tetrabromo-1H benzotriazole were obtained by direct alkylation of 4,5,6,7-tetrabromo-1H benzotriazole with the use of respective alkyl halides in the presence of KOH in methanol, to give a mixture of 1- and 2- isomers, which was separated by flash column chromatography in good yield. The proportion of isomers strongly depended on the reaction time and temperature. 1- and 2-hydroxyethyl and 1- and 2 chloroethyl derivatives of the tetrabromobenzo-triazole were synthesized with the use of 2-bromoethanol and 1-bromo-2-chloroethane respectively as alkylating agents. N-alkylation of this benzotriazole compound enhanced inhibitory activity and selectivity towards the helicase activity of HCV NTPase/helicase. The most active were the 2-methyl, 2-ethyl and 2-propyl derivatives (IC50 approximately 6.5 microM in the presence of DNA as a substrate). Derivatives of the benzotriazole in which hydroxyethyl or chloroethyl replaced the alkyl substituents lost their inhibitory activity. Brominated or methylated benzotriazole N(1) ribosides also did not exert helicase inhibitory activity. Although a number of N(1) and N(2) alkyl derivatives exerted good HCV and WNV helicase inhibitory activity when DNA was used as substrate, the activity was strongly decreased or even disappeared when RNA was used as substrate. The cytotoxicity tests in Vero and HeLa Tat cells showed a substantial decrease of cytotoxicity of N-alkyl derivatives as compared to the parent benzotriazole. PMID- 16245649 TI - Basic biology of tendon injury and healing. AB - Tendon disorders are commonly seen in clinical practice. Their successful treatment is difficult and patients often experience symptoms for prolonged periods of time. At present the aetiology of tendon disorders remains unclear, with several factors having been implicated. An improved understanding of tendon injury and healing is essential to enable focused treatment strategies to be devised. PMID- 16245648 TI - Antiviral and cytostatic evaluation of the novel 6-acyclic chain substituted thymine derivatives. AB - A series of the novel 5-methyl pyrimidine derivatives with an acyclic side chain at the C-6 position were synthesized using lithiation of a 2,4-dimethoxy-5,6 dimethyl pyrimidine and subsequent nucleophilic addition or substitution reactions of the organolithium intermediate thus obtained with acetaldehyde, epichlorhydrine, fluorinated ketones and fluorinated ester. The novel compounds were evaluated for their cytostatic and antiviral activities. Among all the compounds evaluated, two fluorinated acyclic pyrimidine derivatives showed the highest cytostatic activities. The compound containing a 2-hydroxy-3,3,3 trifluoro-1-propenyl side chain exhibited a pronounced effect against breast carcinoma (MCF-7, IC50=8.38 micorg/ml), while the compound with a 2-fluoromethyl 2-acetoxypropyl chain exhibited moderate effect against cervical carcinoma (HeLa, IC50=19.73 microg/ml). PMID- 16245650 TI - Intraductal papillary mucinous neoplasms of the pancreas. AB - BACKGROUND: Intraductal papillary mucinous neoplasm (IPMN) of the pancreas is a new pathological entity. It is diagnosed with increasing frequency. However, its natural history and management are still not well defined. METHODS: A Medline search was undertaken to identify articles using the keywords "intraductal papillary mucinous neoplasms of pancreas", "pancreatic neoplasms", and "pancreatic cyst". Additional papers were identified by a manual search of the references from the key articles. RESULTS: Surgical resection is the only treatment which can produce a cure. The reported overall 5-year survival for IPMN after surgical resection varies from 36% to 77%; for non-invasive IPMN, 77% to 100% and for invasive IPMN, 27% to 60%. The overall recurrence rate was 7% to 43%. IPMN can recur either as disseminated disease or as isolated pancreatic remnant recurrence even after surgical resection with negative margins. CONCLUSIONS: Based on the available evidence, patients with IPMN should undergo complete surgical resection. The extent of pancreatic resection and the intra operative management of resection margins remain controversial. Balancing the risk of recurrence and the morbidity of total pancreatectomy, routine total pancreatectomy for IPMN is not recommended. Total pancreatectomy should only be reserved for patients with resectable but extensive IPMN which involves the whole pancreas. Regular monitoring for disease recurrence is important after surgery as there is a risk of recurrence in both non-invasive and invasive IPMN, and repeat resection for an isolated recurrence in the pancreatic remnant gives good results. PMID- 16245651 TI - Langer's axillary arch: anatomy, embryological features and surgical implications. AB - Langer's arch is identified in up to 7% of axillary explorations; in a three month period we identified three individuals among forty-six patients undergoing axillary surgery with this abnormality. Langer's arch is a muscular-tendinous structure that usually extends from latissimus dorsi to pectoralis major muscle. The purpose of this article is to describe the embryological derivation of this muscular variant from the panniculus carnosus and to define its anatomical features. The clinical implications both symptomatically and as an unexpected finding during axillary dissection, are also discussed. PMID- 16245652 TI - A review of the current management of severe traumatic brain injury. AB - Traumatic brain injury accounts for up to half of trauma related fatalities. This review describes current management practices including pre-hospital care, surgical interventions and various treatment modalities for intracranial hypertension. The lack of class I evidence for the majority of interventions is highlighted. PMID- 16245653 TI - The relationship between severity of obstructive sleep apnoea/hypopnoea syndrome (OSAHS) and lateral cephalometric radiograph values: a clinical diagnostic tool. AB - BACKGROUND: This prospective cross-sectional cohort study examined the relationship between radiographic anatomy and the severity of OSAHS. The severity of OSAHS can be measured subjectively in terms of the Epworth scale and objectively in terms of the apnoea/hypopnoea Index (AHI). METHODS: 121 lateral cephalometric radiographs were traced under uniform conditions and a series of 56 landmarks identified, from which 48 angular and linear measurements were made. Significant changes occurred when comparison of these measurements with the severity of OSAHS were made. RESULTS: Body mass index (BMI), the maxillary mandibular planes angle (MMPA) and the linear measurement between points 7 and 12, (the pharyngeal dimension measured from the tip of the soft palate to the corresponding horizontal point on the posterior pharynx), increased significantly with increasing severity of OSAHS as measured by the Epworth score. Overjet, lower lip length, and the distance from the hyoid bone to a point B on the mandible all increased significantly with increasing severity of OSAHS, as measured by the AHI. The hyoid bone was found to rotate counter clockwise as the severity of OSAHS increased, as a result the distance between the most anterior superior point on the hyoid bone and the maxillary plane was seen to decrease as severity of OSAHS increased in terms of AHI. CONCLUSIONS: Some radiographic anatomical features show significant change as the severity of OSAHS increases and these features could be used in the identification of patients who have severe OSAHS. The Logit equation derived from the findings of this study may also be a useful clinical tool in predicting the likelihood of a subject suffering from severe OSAHS. PMID- 16245654 TI - Long-term morbidity following short-course, pre-operative radiotherapy and total mesorectal excision for rectal cancer. AB - AIM: Short-course pre-operative radiotherapy (SCPRT) and total mesorectal excision (TME) have been shown to reduce the rate of relapse and improve survival in patients with rectal cancer. Concerns about morbidity have limited its application in some centres. The aim of this study was to assess long-term toxicity of pre-operative RT in patients with rectal cancer. METHODS: Permanent toxicity was assessed in 20 patients who were alive >12 months after pre operative RT (25Gy/ 5#), using a validated LENT SOMA scoring system. Ten patients with rectal cancer who had been operated on >12 months ago, but had not received pre-operative RT were assessed similarly. RESULTS: Patients who had received SCPRT appeared to have a significantly higher rectal toxicity, urinary and male sexual dysfunction, compared with the controls. No significant difference was noted in female sexual dysfunction. CONCLUSION: The combination of pre-operative RT and TME appears to be associated with significant long-term toxicity. Better methods of staging the tumours may improve selection of patients for SCPRT. PMID- 16245655 TI - Low incidence of retained common bile duct stones using a selective policy of biliary imaging. AB - AIM: Biliary imaging has decreased since the advent of laparoscopic cholecystectomy. This study aimed to examine the incidence of retained common bile duct (CBD) stones, using a selective policy of biliary imaging. METHODS: A prospective computerised database was used to study patients who underwent laparoscopic cholecystectomy by one surgeon over 36 months. Two hundred and thirty-five patients (191 female, 44 male) were operated on. Ages ranged from 15 to 82 years (mean 47 years). Follow-up periods ranged from six to 39 months (mean 23.3 months). Selection for pre-operative endoscopic retrograde cholangiopacreatogram (ERCP), intraoperative cholangiography (IOC) or neither depended on liver function tests (LFTs) and CBD diameter on ultrasound prior to surgery. If LFTs were persistently raised and/or the CBD was dilated on ultrasound, patients underwent pre-operative ERCP. If LFTs were raised, and returned to normal promptly, IOC was performed. RESULTS: Of 235 patients, 26 had pre-operative ERCP, 62 had IOC and 157 had neither. Ten patients had both ERCP and IOC. To date, one patient (0.43%) has radiologically-proven unsuspected retained CBD stones; these were successfully treated with ERCP. CONCLUSION: Selective biliary imaging in patients undergoing laparoscopic cholecystectomy is safe. This process identifies the majority of patients with ductal calculi and minimises the need for unnecessary ERCP and peroperative cholangiography. PMID- 16245656 TI - Primary malignant melanoma of the colon in an oculocutaneous albino. AB - We describe the case of an elderly albino man who presented with the typical features of a caecal malignancy and underwent a right hemi-colectomy. Histological assessment subsequently revealed the tumour to be a primary malignant melanoma of the colon. This unexpected diagnosis raises interesting questions regarding the embryology and aetiology of this rare tumour and the anticipated prognosis. To the best of our knowledge, this is the first reported case of a primary colonic melanoma associated with oculocutaneous albinism. PMID- 16245657 TI - Re: Same admission colostomy closure: a prospective, randomized study in selected patient groups. Surgeon 2004; 3(1):11-14. PMID- 16245658 TI - Re: Meshikhes A-W. N. Daflon for haemorrhoids: a prospective, multi-centre observational study. Surg JR Coll Surg Edinb Irel 2004;2(6): 355-38. PMID- 16245659 TI - Controversies in canine disc disease. PMID- 16245660 TI - Canine glomerulonephritis: new thoughts on proteinuria and treatment. AB - Glomerular disease in the dog is not only a common form of renal disease but also an important cause of chronic renal failure. The presence of immune complexes in glomerular capillary walls is a major cause of canine glomerular disease and is commonly referred to as glomerulonephritis. Leakage of plasma proteins, principally albumin, across the damaged glomerular capillary walls results in persistent proteinuria--the clinicopathological hallmark of glomerulonephritis. Recent evidence suggests that, in addition to being a marker of disease, persistent proteinuria is associated with progressive glomerular and tubulointerstitial lesions and loss of additional nephrons. Perhaps the best treatment for glomerulonephritis is the identification and correction of any underlying inflammatory, immune-mediated or neoplastic disease that results in the deposition or formation of glomerular immune complexes. In cases of idiopathic glomerulonephritis, angiotensin-converting enzyme inhibitors have been shown to decrease proteinuria and potentially slow disease progression. PMID- 16245661 TI - Partial percutaneous discectomy for treatment of thoracolumbar disc protrusion: retrospective study of 331 dogs. AB - OBJECTIVES: To determine retrospectively the prognosis and outcome for dogs diagnosed with thoracolumbar intervertebral disc disease treated with partial percutaneous discectomy (PPD). METHODS: Three hundred and thirty-one dogs presenting with symptoms of thoracolumbar intervertebral disc disease from 1998 to 2003 were treated with PPD. Diagnosis and location of intervertebral disc disease was confirmed by clinical examination, radiography, myelography and magnetic resonance imaging. PPD was performed via fluoroscopy-guided removal of a 5 mm bore cylinder out of the central intervertebral space. RESULTS: Clinical success after surgery was achieved in 159 (88.8 per cent) grade II to IV patients and 58 (38.2 per cent) grade V patients. The mean (sd) time from percutaneous discectomy to first improvement was 8.3 (13.2) days. CLINICAL SIGNIFICANCE: The PPD approach to the thoracolumbar spine involves minor trauma (yielding rapid recovery) and less pain, and produces results comparable with open fenestration. Consequently, this simple minimal invasive technique can be recommended as an alternative to the technique of fenestration and can be easily performed in addition to open surgical decompression techniques or prophylactically. However, it is not a replacement for surgical treatment in dogs with thoracolumbar disc disease that require removal of disc fragments causing spinal cord or nerve root compression. PMID- 16245662 TI - Thoracolumbar disc extrusion associated with extensive epidural haemorrhage: a retrospective study of 23 dogs. AB - OBJECTIVES: To describe the clinical features and outcome in dogs suffering from thoracolumbar disc extrusion associated with extensive epidural haemorrhage (DEEH) and treated with extensive hemilaminectomy (from three to seven vertebrae). METHODS: The records of 23 dogs with surgically confirmed DEEH were reviewed retrospectively. RESULTS: All cases were characterised by rapid progression to severe neurological dysfunction (grade III, V and VI). Myelography was performed in 21 cases and showed an absence (16 cases) or attenuation (five cases) of contrast medium column along three to seven vertebrae. In two dogs, magnetic resonance imaging was accurate in confirming extradural compression due to disc material and haemorrhage, determining the extent of compression and side of the lesion. All cases were treated surgically with extensive hemilaminectomy involving all the compressed spinal segments. Twenty-one dogs (91 per cent) recovered and regained ambulatory function. Two dogs, without deep pain perception before surgery, did not improve. A two-year follow-up history was available for 15 dogs. Disc extrusion recurred in two dogs (9 per cent), two and 20 months after surgery. CLINICAL SIGNIFICANCE: Extensive hemilaminectomy can adequately decompress the spinal cord after DEEH and may produce a recovery and recurrence rate similar to thoracolumbar disc extrusion not complicated by extensive epidural haemorrhage. PMID- 16245663 TI - Identifying risk factors associated with MRSA infection in companion animals. PMID- 16245664 TI - Effect of various thoracic radiographic projections on the appearance of selected thoracic viscera. AB - OBJECTIVES: To assess the effect of different radiographic projections on thoracic width, as well as position and visibility of the trachea, principal bronchi, cardiac silhouette, aorta, caudal vena cava (CVC) and oesophagus. METHODS: Right lateral recumbency (RLR), left lateral recumbency (LLR), dorsoventral (DV) and ventrodorsal (VD) thoracic radiographs of 42 dogs were reviewed retrospectively. RESULTS: In 78 per cent of cases the thoracic width was significantly larger on the VD projection than on the DV projection. The angle of divergence formed by the principal bronchi was significantly larger on the VD projection than on the DV projection in 80 per cent of dogs. A cardiac silhouette bulge at 1 to 2 o'clock was apparent on the VD projection in 22 per cent of dogs but was never seen on DV projections. The descending aorta was more visible at the 4 to 5 o'clock cardiac silhouette level on the DV projection and laterally at the T8 level on LLR projections. The CVC was better seen on VD and LLR projections. The oesophagus was visible as a soft tissue opacity in LLR in large dogs with normal thoracic conformation in 35 per cent of cases. CLINICAL SIGNIFICANCE: The DV projection appears to be more reliable for assessing the cardiac silhouette, the descending aorta and the angle of divergence of the principal bronchi. The VD projection should be considered for evaluating the CVC. LLR should be used for assessing the descending aorta and CVC. PMID- 16245665 TI - Chronic axillary wound repair in a cat with omentalisation and omocervical skin flap. AB - A mature, neutered male domestic cat was presented with a chronic axillary wound of unknown origin and at least three years' duration. The diagnostic investigation included screening tests for feline leukaemia virus and feline immunodeficiency virus, tissue culture and histological examination. No underlying aetiology or perpetuating cause could be identified. An omental flap was created, passed via a subcutaneous tunnel, and packed into the wound site after excision of all chronic granulation tissue. The skin defect was closed using an omocervical axial pattern skin flap. A small area of the distal edge of the flap became necrotic but the defect healed by second intention. To the author's knowledge, this is the first clinical report of the use of an omocervical skin flap for repair of a chronic axillary wound in a cat. This flap offers a useful alternative where the use of a thoracodorsal axial pattern flap is not possible due to the extent of the lesion. PMID- 16245666 TI - Urethral transitional cell carcinoma in a cat. AB - A 15-year-old, male neutered cat was referred for investigation of dysuria. A retrograde urethrography was performed which showed two space-occupying masses within the lumen of the mid-to-proximal urethra. Exploratory coeliotomy revealed two urethral masses. Segmental urethrectomy was performed to resect the mass, and the lower urinary tract was reconstructed by vesico-urethral anastomosis. Histopathology showed the mass to be a transitional cell carcinoma with incomplete surgical margins. Tumour regrowth was suspected when dysuria was found approximately 318 days after surgery. Clinical signs were palliated by radiation using weekly fractions of 6 Gy for three weeks. The cat died of unknown causes 386 days postoperatively. PMID- 16245668 TI - Talking sense. PMID- 16245667 TI - What is your diagnosis? Gastrocnemius enthesiopathy. PMID- 16245669 TI - Caring to learn? Getting young mothers back into education. PMID- 16245671 TI - World Mental Health Day 2005. PMID- 16245670 TI - The economics of sexual health. PMID- 16245672 TI - Partners in Health. PMID- 16245673 TI - How can I quit passive smoking? AB - Passive smoking is a topical subject and there is a concerted movement to increase public understanding of the dangers of passive smoking. Although it looks likely that smoking could be banned in public places in the UK by the year 2008, it will still be difficult to enforce smoking bans in the last bastion for the smokers--their homes. Many smokers are aware of the risk their smoking causes their families through passive smoking but do not realise that the only true method for them to reduce exposure for their family is to smoke outside the home. This is partly because of a lack of understanding of the behaviour of environmental tobacco smoke and how smoking in restricted areas alone will not eliminate passive smoking for other family members in their homes. PMID- 16245674 TI - At home with clients: a study of health visiting expertise. AB - This paper describes the methods and some findings of a qualitative study of domiciliary health visiting practice undertaken in central Scotland between 2000 and 2002. The study aimed to describe the health visiting expertise used in assessing and responding to client needs during home visits. An unusual combination of methods was used, including simulated visits to two actress clients, post-simulation interviews and subsequent observation of participants' actual practice. This combination of methods produced rich and complex data about this area of practice. The study findings which are described in this paper relate to the nature and range of client needs addressed during visits, to the two distinct practitioner approaches which were identified and to how these two approaches were related to the main focus of the visits. Other findings are outlined and the study's limitations and recommendations are briefly addressed. PMID- 16245675 TI - Pilot trial of an age-paced parenting newsletter. AB - Supporting parents in the first three years of a child's life has the potential to produce successful outcomes. Present government initiatives such as Sure Start focus on this age group. An American educational intervention, in the style of a monthly newsletter, was adapted for use in the UK for parents of young children. Topics were presented in an easy-to-read format and focused on infant emotional development, parent interaction and play. Newsletters, called Baby Express were posted at monthly intervals to the family home providing age-paced information which could meet the specific needs of parents at that stage of their child's life. The aim of the study was to determine the applicability of the newsletter to UK parents and evaluate their satisfaction. Sixty home-based interviews were conducted and 95 per cent of mothers reported reading all or part of the newsletter. Changes in parenting style were spontaneously reported by 28 per cent of mothers. This study found that an aged-paced parenting newsletter was an acceptable and useful method of supporting parents in the early months of a child's life and promotes positive changes in parenting behaviour. PMID- 16245676 TI - Vitamins, minerals and supplements: part two. AB - Vitamins and minerals are organic food substances found only in plants and animals and are essential to the normal functioning of the body. Although only required in small amounts, as previously discussed in the past decade there has been an increased use of vitamin, mineral, herbal and nutritional supplements in the general population. While deficiencies in such nutrients can be harmful to health, conflicting claims have been made about the health benefits of such supplementation. In the second of an occasional series on vitamins, minerals, and supplements, JUNE THOMPSON gives an overview of the role that water-soluble vitamins play in the health of the individual, including their functions, and the potential impact of any deficiency of these. PMID- 16245677 TI - Mistakes--a stick to beat health workers? PMID- 16245678 TI - [Adult stem cells: who are they, what do they do?]. AB - Fascinating and provocative findings have shaken the stem cell research field in recent years. One unexpected discovery is the identification of stem/progenitor like cells in many tissues with slow cellular turnover, such as heart, kidney, muscle and brain. Cells with high proliferative capacity and multilineage differentiation potential have also been described in bone marrow, although their existence needs to be confirmed. Both cell types may prove to have therapeutic potential, but research on their use for tissue repair has been rather disappointing. In addition, serious doubts have been raised concerning the transdifferentiation potential of hematopoietic stem cells, underlining the need for care when interpreting findings that question long-established concepts. PMID- 16245679 TI - [Stem cells and neural repair]. AB - The presence of stem cells in the central nervous system of adult rodents has been suspected some forty years ago. However, it is only since two decades that the ability of those cells to give rise to neurons has been demonstrated in two regions of the CNS, the dentate gyrus of the hippocampus and the olfactory bulb. It is only recently that stem cells have been identified in the hippocampus of adult Humans. Stem cells have been transplanted in animal models of Nervous pathologies, to compensate for a deficit in neurotransmitters (Parkinson's disease) or of trophic factors (Spinal cord injury) with encouraging but not decisive results. Future progresses are expected from the us of intrinsic stem cells whose fate could be controlled thanks to the modern tools of gene therapy. PMID- 16245680 TI - [Role of stem cells in cardiac repair]. AB - The use of stem cells for cardiac repair is based on their potential to become cardiomyocytes and thereby to restore the functional capacity of the failing heart. The concept of transdifferentiation, by which adult stem cells adopt the fate of the cells they are intended to replace, generated enormous enthusiasm, but it is increasingly recognized that the plasticity of these cells is rather limited. This is particularly the case of skeletal myoblasts, which remain committed to their myogenic lineage. Likewise, conversion of bone marrow-derived cells into cardiomyocytes is, at most, an exceptional and quantitatively limited event. These limitations do not preclude the possibility that adult stem cells could have protective effects on left ventricular function, possibly through a paracrine action. Tissue-resident cardiac stem cells were recently identified but significant hurdles will have to be overcome if they are to be used for therapeutic purposes. Consequently, much hope is currently being placed in embryonic stem cells which, provided they are appropriately precommitted during culture, can differentiate into cardiomyocytes. Their subsequent electromechanical integration into the recipient myocardium can contribute to the repair of the damaged heart. PMID- 16245681 TI - [Liver repopulation strategies]. AB - Hepatocytes have the unique capacity to self-renew and repair the liver ad integrum when stimulated to proliferate by liver injury. However, transplantation of isolated hepatocytes is usually not sufficiently efficient for therapeutic purposes. We conferred a survival advantage on transplanted hepatocytes and showed that they were able to repopulate almost the entire mouse liver after repeated injury. In contrast, we found that bone marrow stem cell transdifferentiation was inadequate for therapeutic liver regeneration. Current data on liver stem cells will be discussed. PMID- 16245682 TI - [Contribution of stem cells to renal repair after ischemia/reperfusion]. AB - Repair of inflammatory and/or ischemic renal injury involves endothelial, mesangial and epithelial regeneration. These structures may be rebuilt by resident progenitor cells and bone marrow-derived stem cells. Resident progenitor cells in adult kidney have not yet been conclusively identified. They are likely to be slowly cycling cells located mainly in the outer medulla and renal papilla. In glomerulonephritis with mesangiolysis, mesangial regenera- tion involves progenitor cells migrating from the juxtaglomerular apparatus and also bone marrow-derived cells. In acute ischemic renal failure, epithelial regeneration of proximal tubules results from the migration, proliferation and differentiation of resident progenitor cells; bone marrow-derived cells may play an accessory role. Molecular mechanisms underlying these repair processes could be targets for new therapeutic approaches. PMID- 16245683 TI - [Embryonic stem cells and epidermal commitment]. AB - Embryonic stem (ES) cells can be cultured indefinitely, differentiated into many cell types in vitro, thus providing a potentially unlimited supply of cells for cell-based therapy. We recently reported the efficient derivation of ectodermal and epidermal cells from murine ES cells. These differentiated ES cells are able to form, in culture, a multilayered epidermis coupled with an underlying dermal compartment, similar to native skin. This model demons- trates that ES cells have the potential to recapitulate the reciprocal instructive ectodermal-mesodermal commitments, characteristic of embryonic skin formation, clarifies the role of the morphogen BMP-4 in the binary neuroectodermal choice and provides a powerful tool for the study of molecular mechanisms controlling skin development and multipotent epidermal stem cell properties. Its potential for cutaneous cell therapy and dermatocosmetological applications is discussed. PMID- 16245684 TI - [Non-invasive coronary artery imaging using multislice computed tomography]. AB - Coronary atherosclerotic disease is the leading cause of mortality in industrialized countries. The disease can remain silent for years, being diagnosed only after acute myocardial infarction in about 50% of cases. Coronary angiography is the reference standard for coronary imaging. However, it is invasive and costly, requiring intra-arterial catheterization. In addition, coronary angiography does not visualize the arterial wall, but only the lumen. A reliable non-invasive imaging technique showing changes in both the arterial wall and the lumen would be highly useful, especially for early diagnosis. Recently, multislice computed tomography (CT) was shown to be capable of visualizing not only the coronary arteries (lumen and wall), but also the cardiac muscle, with high spatial resolution. Multislice CT provides high-quality 3D images of coronary arteries. Initial results with 4, 16 and now 64-slice CT, by comparison with conventional angiography, are very promising, and multislice CT technology is advancing rapidly. Non-invasive visualization of the coronary arteries with accurate detection of stenosis is now possible in practice using ECG-gated 16 slice CT Multislice CT can also detect non stenotic coronary plaque. Finally, visualization of cardiac muscle by multislice CT makes it possible to detect myocardial ischemia. PMID- 16245685 TI - [Evolution of musculoskeletal ultrasonography]. AB - Sonography is a safe, inexpensive and readily accessible technique which has acquired a major place in musculoskeletal imaging over the past two decades. Thanks to recent technical innovations such as high-definition multifrequency probes, compound effect, use of harmonic frequencies, power Doppler, and extended field of view, today's sonographic images are extremely precise. At the same time, based on magnetic resonance imaging (MRI) and computed tomography (CT) acquisitions, the sonographic anatomy has become broader and more detailed, and the sonographic semiology is more precise and reliable. In this article the authors evaluate current uses of diagnostic and interventional sonography in musculoskeletal disorders. PMID- 16245686 TI - [The possible place of autologus cell therapy in facioscapulohumeral muscular dystrophy]. AB - Facioscapulohumeral dystrophy (FSHD), one of the most common forms of muscular dystrophy, derives its name from the patients' selective, often asymmetric clinical distribution of muscle weakness. Interestingly, affected and non affected areas can coexist in the same patient for many years. The molecular hallmark is total deletion of the subtelomeric D4Z4 repeat on chromosome 4q. There is no specific treatment. Gene therapy is unlikely to be feasible, as no alterations have been found in the genes located in this subtelomeric region. Muscular dystrophies are characterized by the coexistence of genetically induced muscle degeneration and compensatory muscle regeneration by myoblast proliferation from satellite cells; muscle weakness and atrophy appears when this mechanism is overwhelmed. Cell therapy with autologous myoblasts would, in theory, be a simple way of boosting the regenerative process and of preventing or delaying muscle degeneration. This approach might also avoid the use of toxic immunotherapies. By using a recent very-high-yield cell culture method, we analyzed the proliferation and differentiation of myoblasts obtained from FSHD patients, both ex vivo and in vivo (by intramuscular injection to immunodeficient mice). Myoblasts were obtained by muscle biopsy from five FSHD patients harboring the D4Z4 deletion. We selected the vastus lateralis muscle, which exhibited no clinical, radiological or pathological signs of dystrophy. The growth characteristics of these cells were compared with those of cells from normal control muscles, based on the culture yield, phenotypic characterization with anti-CD56 and anti-desmin antibodies, and the capacity for differentiation (myotube production in vitro and human dystrophin expression one month after injection to Rag2 immunodeficient mice). Patients' cells recovered from 1 g of muscle biopsy specimen resembled control cells in terms of their growth kinetics, culture yield, and capacity to differentiate and produce mature muscle cells. These results indicate that myoblasts taken from unaffected muscle of patients with FSHD warrant testing in a human cell therapy trial. PMID- 16245687 TI - [Gamete and embryo protection against oxidative stress during medically assisted reproduction]. AB - So-called oxidative stress is due to the generation of reactive oxygen species (ROS), the most important of which are the superoxide anion (O2-*), hydrogen peroxide, and the hydroxyl radical OH*. ROS can damage gametes and embryos, through DNA (and RNA) fragmentation and membrane lipid peroxidation, potentially resulting in miscarriage. ROS are produced both physiologically (by oxidative metabolism) and in response to environmental factors such as oxygen, light, and traces of divalent cations. The body possesses several defense mechanisms against ROS, including antioxidant enzymes such as superoxide dismutase (SOD), catalase, glutathione peroxidase, and also small molecules like vitamins (A, C and E), glutathione and pyruvate. These different systems generally act in concert. Embryonic cells also possess DNA repair machinery. As oxidative stress is one of the main sources of embryo loss in vitro, it is important to protect both gametes and embryos against these insults during medically assisted reproduction. PMID- 16245688 TI - [Ester derivatives of nucleoside inhibitors of reverse transcriptase: II. Molecular systems for combined therapy with 3'-azido-3'-deoxythymidine and 2',3' didehydro-3'-deoxythymidine]. AB - The methods of synthesis of conjugates of anti-HIV nucleosides with various compounds, such as protease inhibitors, peptides, polysaccharides, and bicyclames, are considered; they are designated for the combined therapy of HIV. PMID- 16245689 TI - [Theoretical conformational analysis in the determination of productive conformations of substrates for acetylcholinesterase and butyrylcholinesterase]. AB - All the equilibrium conformations of 34 analogues of acetylcholine (ACh) with the general formula R-C(O)O-Alk-N+(CH3)3 are calculated by the method of molecular mechanics. In the series R-C(O)O-(CH2)2-N+(CH3)3, a reliable correlation is found between the molecular volume of the substrate and the rate of its hydrolysis by acetylcholinesterase (AChE); the absence of such a correlation is demonstrated for butyrylcholinesterase (BChE). Theoretical conformational analysis confirms that the completely extended tt conformation of ACh is productive for the hydrolysis by AChE, which agrees with the results of X-ray analysis of AChE. AChE is shown to hydrolyze only those substrates that form equilibrium conformers compatible in the mutual arrangement of trimethylammonium group, carbonyl carbon, and carbonyl oxygen with the tt conformation of ACh; in this case, the rate of substrate hydrolysis depends on the total population of these conformers. A reliable correlation was found between the population of the semifolded (tg-) conformation of the choline moiety of substrate molecules and the rate of their BChE hydrolysis. In a series of CH3-C(O)O-Alk-N+(CH3)3, the rate of BChE hydrolysis is demonstrated to depend on the total population of conformations compatible in the mutual arrangement of functionally important atoms with the tg- conformation of ACh. The tg- conformation of ACh is concluded to be productive for BChE hydrolysis. Similar orientations of the substrate molecules relative to the catalytic triads of both AChE and BChE are proven to coincide upon the substrate productive sorption in their active sites. It is hypothesized that the sorption stage is rate-limiting in cholinesterase hydrolysis and the enzyme hydrolyzes the ACh molecule in its energetically favorable conformation. PMID- 16245690 TI - [A new Escherichia coli strain producing human tumor necrosis factor]. AB - An Escherichia coli strain producing human tumor necrosis factor (TNF-alpha) was obtained using a semisynthetic gene partially optimized in respect of codon composition and a phage T7 promoter. The expression product was accumulated in cells as inclusion bodies in a yield of 50-70 mg/l of culture medium. The recombinant TNF-alpha in the form of inclusion bodies was used for immunization of rats to give a polyclonal antiserum. The resulting antibodies were specific under the immunoblotting conditions to the antigen used for the immunization. A dilution-based refolding procedure was developed; it provided a yield of soluble protein exceeding 85%. PMID- 16245691 TI - [Epitope mapping of the recombinant movement protein of the tobacco mosaic virus using monoclonal antibodies]. AB - The movement protein (MP) of the tobacco mosaic virus (TMV) provides the intercellular transport of the viral RNA through plasmodesmata. The MP fulfills its function while interacting with host cell factors over the whole path of its intracellular movement from the subcellular site of its synthesis to the plasmodesmata of cellular walls. The MP conformation during its intracellular movement and fulfillment of the transport function still remains unknown. In this study, we describe the preparation of murine monoclonal antibodies (MAs) to TMV MP and mapping of the MP epitopes. Stable hybridoma lines that produce MAs to the partially denatured recombinant MP (MPr) were obtained. MAs were tested by immunoblotting and ELISA with the use of deletion variants of MPr. The epitopes of TMV MPr that recognize specific MAs were determined. PMID- 16245692 TI - [Theoretical study of the structure of adenosine deaminase complexes with adenosine analogues: I. Aza-, deaza- and isomeric azadeazaanalogues of adenosine]. AB - The conformational models of the active site of adenosine deaminase (ADA) and its complexes in the basic state with adenosine and 13 isosteric analogues of the aza, deaza, and azadeaza series were constructed. The optimization of the conformational energy of the active site and the nucleoside bound with it in the complex was achieved in the force field of the whole enzyme (the 1ADD structure was used) within the molecular mechanics model using the AMBER 99 potentials. The stable conformational states of each of the complexes, as well as the optimal conformation of the ADA in the absence of ligand, were determined. It was proved that the conformational state that is close to the structure of the ADA complex with 1-deazaadenosine (1ADD) known from the X-ray study corresponds to one of the local minima of the potential surface. Another, a significantly deeper minimum was determined; it differs from the first minimum by the mutual orientation of side chains of amino acid residues. A similar conformational state is optimal for the ADA active site in the absence of the bound ligand. A qualitative correlation exists between the values of potential energies of the complexes in this conformation and the enzymatic activity of ADA toward the corresponding nucleosides. The dynamics of conformational conversions of the active site after the binding of substrate or its analogues, as well as the possibility of the estimation of the inhibitory properties of nucleosides on the basis of calculations, are discussed. PMID- 16245693 TI - [A structural and thermodynamic analysis of novatrone and flavin mononucleotide heteroassociation in aqueous solution by 1H NMR spectroscopy]. AB - A heteroassociation of antitumor antibiotic novatrone (NOV) and flavin mononucleotide (FMN) in aqueous solution was studied by one- and two-dimentional 1H NMR spectroscopy (500 MHz) to elucidate the molecular mechanism of the possible combined action of the antibiotic and vitamin. The equilibrium reaction constants, induced proton chemical shifts, and the thermodynamic parameters (deltaH and deltaS) of the NOV and FMN heteroassociation were determined from the concentration and temperature dependences of proton chemical shifts of the aromatic molecules. The most favorable structure of the 1 : 1 NOV-FMN complex was determined by both the method of molecular mechanics (X-PLOR software) and the induced proton chemical shifts of the molecules. An analysis of the results suggests that the NOV-FMN intermolecular complexes are mainly stabilized by stacking interactions of their aromatic chromophores. An additional stabilization is possible due to intermolecular hydrogen bonds. It was concluded that the aromatic molecules of vitamins, in particular, FMN, can form energetically favorable heterocomplexes with aromatic antitumor antibiotics in aqueous solutions, which could result in a modulation of their medical and biological action. PMID- 16245694 TI - [A synthesis of heteroaromatic S- and N-beta-glycosides of N-acetylglucosamine under phase transfer conditions]. AB - The use of crown ethers for a phase transfer-catalyzed synthesis of heteroaromatic glycosides of N-acetylglucosamine was studied. The solid-liquid system and the catalysis by 15-crown-5 were found to provide for both a 100% conversion of the alpha-D-glucosaminyl chloride peracetate and a high reaction rate. The interaction of alpha-D-glucosaminyl chloride peracetate and oxadiazole and triazole mercapto derivatives capable of thiol-thione tautomerism carried out at room temperature in acetonitrile in the presence of anhydrous potassium carbonate and crown ethers was shown to lead to both S- and N-glycosides. The structures of the compounds synthesized were confirmed by X-ray analysis and 13C and 1H NMR spectroscopy. PMID- 16245696 TI - [22,23-epoxides of sitosterol and related 7-oxygenated delta5-sterols]. AB - (22S,23S)-22,23-Epoxysitosterol, (22R,23R)-22,23-epoxysitosterol, (22S,23S)-22,23 epoxy-7-ketositosterol, (22R,23R)-22,23-epoxy-7-ketositosterol, (22S,23S)-22,23 epoxy-7alpha-hydroxysitosterol, (22R,23R)-22,23-epoxy-7alpha-hydroxysitosterol, (22S,23S)-22,23-epoxy-7beta-hydroxysitosterol, and (22R,23R)-22,23-epoxy-7beta hydroxysitosterol were synthesized. Their 1H and 13C NMR and the mass spectra of their trimethylsilyl derivatives were studied. PMID- 16245695 TI - [Steroid compounds from far Eastern starfishes Henricia aspera and H. tumida]. AB - Six new natural compounds were isolated from two Far Eastern starfish species, Henricia aspera and H. tumida, collected in the Sea of Okhotsk. Two new glycosylated steroid polyols were obtained from H. aspera: asperoside A and asperoside B, which were shown to be (20R,24R,25S)-3-O-(2,3-di-O-methyl-beta-D xylopyranosyl)-24-methyl-5alpha-cholest-4-ene-3beta,6beta,8,15a,16beta,26-hexaol and (20R,24R,25S,22E)-3-O-(2,4-di-O-methyl-beta-D-xylopyranosyl)-24-methyl-5alpha cholest-22-ene-3beta,4beta,6beta,8,15alpha,26-hexaol, respectively. Two other glycosylated polyols, tumidoside A, with the structure elucidated as (20R,22E)-3 O-(2,4-di-O-methyl-beta-D-xylopyranosyl)-26,27-di-nor-24-methyl-5alpha-cholest-22 ene-3beta,4beta,6beta,8,15alpha,25-hexaol, and tumidoside B, whose structure was elucidated as (20R,24S)-3-O-(2,3-di-O-methyl-beta-D-xylopyranosyl)-5alpha cholestan-3beta,4beta,6beta,8,15alpha,24-hexaol, were isolated from the two starfish species. (20R,24S)-Salpha-Cholestan-3beta,6beta,15alpha,24-tetraol and (20R,24S)-5alpha-cholestan-3beta,6beta,8,15alpha,24-pentaol were identified only in H. tumida. The known monoglycosides henricioside H1 and laeviuscolosides H and G were also identified in both species. PMID- 16245697 TI - [Effect of substituents on photochemical and biological properties of 13,15-N cycloimide derivatives of chlorin p6]. AB - The effect of electron-accepting substituents in position 3 of the chlorine p6 macrocycle in neutral and carboxyl-containing negatively charged cycloimide derivatives of chlorin p6 (CIC) on the photochemical and biological properties of these photosensitizers was studied. A relationship between the structure and properties of CICs was analyzed on the basis of information on their photoinduced cytotoxicity, efficiency of the generation of reactive oxygen species, photostability, intracellular localization, quantitative parameters of accumulation in cells, and cellular pharmacokinetics. It was shown that these compounds can be used for the development of photosensitizers with intense light absorption at 740 nm, controlled intracellular localization, and a high photodynamic activity toward tumor cells. PMID- 16245698 TI - [Synthesis of lupinine derivatives of flavonoids]. AB - The isomerization and oxidative cyclization of 2-hydroxy-4-lupinylchalcones leads to the corresponding 7-lupinylflavanones and 7-lupinylflavones. PMID- 16245699 TI - [Synthesis of two new acetanilide derivatives and their effect on the serum antioxidant vitamins (A, E, and C) and the MDA level in rats]. AB - Acetanilide derivatives, 2,2'-thiobis[N-(4-nitrophenyl)acetamide] and 2,2' thiobis[N-(4-chlorophenyl)acetamide], were synthesized and characterized. They were shown to cause a considerable oxidative stress in rats. PMID- 16245700 TI - [An Escherichia coli strain producing a leaderless mRNA from the chromosomal lac promoter]. AB - A special Escherichia coli strain capable of producing a leaderless lacZ mRNA from the chromosomal lac promoter was constructed to study the mechanism of leaderless mRNA translation. The translation efficiency of this noncanonical mRNA is very low in comparison with the canonical cellular templates, but it increases by one order of magnitude in the presence of chromosomal mutations in the genes encoding the ribosomal S1 and S2 proteins. The new strain possesses obvious advantages over the commonly used plasmid constructs (first of all, due to the constant dosage of lacZ gene in the cell) and opens possibilities for investigation of the specific conditions for leaderless mRNA translation in vivo using molecular genetic approaches. PMID- 16245701 TI - Distribution of salmonella strains in farrow-to-finish pig herds: a longitudinal study. AB - The aims of this study were to investigate patterns of Salmonella shedding in finishing pigs and to study the role of the sow in the transmission of Salmonella to her offspring. In each of the three herds (A, B, and C), one cohort of sows (n = 34, n = 40, n = 32, respectively) together with three piglets of their offspring (n = 102, n = 120, n = 96, respectively) were selected. Individual fecal and blood samples were taken from the sows at different times during one production cycle and from the piglets from weaning until slaughter. At slaughter, contents from the jejunum, colon, and mesenteric lymph nodes were collected. Fecal samples, as well as the jejunum, colon, and mesenteric lymph node samples collected at slaughter, were submitted to a qualitative Salmonella analysis. Isolates were characterized by random amplified polymorphic DNA, and if necessary, further characterization was done by pulsed-field gel electrophoresis. In herds A and B, Salmonella shedding began in the nursery. A significant increase in the number of Salmonella shedders was seen after transferring pigs to the growing unit in herd B (P = 0.003) and to the finishing unit in herds A (P < 0.001) and B (P = 0.013). None of the fattening pigs in herd C were shedding Salmonella. This study reveals that transferring pigs is an important trigger to induce Salmonella shedding, leading to horizontal spread. Direct transmission of Salmonella from the sows to their piglets could not be demonstrated, but the similarities between the isolates found in the sows and those found during the nursery and finishing periods and at slaughter suggested indirect transmission. PMID- 16245702 TI - Multidrug-resistant Klebsiella pneumoniae isolated from farm environments and retail products in Oklahoma. AB - Multidrug-resistant enteric bacteria were isolated from turkey, cattle, and chicken farms and retail meat products in Oklahoma. Among the isolated species, multidrug-resistant Klebsiella pneumoniae was prevalently isolated from most of the collected samples. Therefore, a total of 132 isolates of K. pneumoniae were characterized to understand their potential roles in the dissemination of antibiotic-resistance genes in the food chains. Multidrug-resistant K. pneumoniae was most frequently recovered from a turkey farm and ground turkey products among the tested samples. All isolates were resistant to ampicillin, tetracycline, streptomycin, gentamycin, and kanamycin. Class 1 integrons located in plasmids were identified as a common carrier of the aadA1 gene, encoding resistance to streptomycin and spectinomycin. Production of beta-lactamase in the K. pneumoniae isolates played a major role in the resistance to beta-lactam agents. Most isolates (96%) possessed bla(SHV1). Five strains were able to express both SHV-11 (pI 6.2) and TEM-1 (pI 5.2) beta-lactamase. Transfer of these antibiotic resistance genes to Escherichia coli was demonstrated by transconjugation. The bacterial genomic DNA restriction patterns by pulsed-field gel electrophoresis showed that the same clones of multidrug-resistant K. pneumoniae remained in feathers, feed, feces, and drinking water in turkey environments, indicating the possible dissemination of antibiotic-resistance genes in the ecosystem and cross contamination of antibiotic-resistant bacteria during processing and distribution of products. PMID- 16245704 TI - Correlation of eggshell strength and Salmonella enteritidis contamination of commercial shell eggs. AB - Shell quality has been identified as a heritable trait that can be manipulated by genetic selection. Previous research has concluded that many methods of determining shell quality produce variable results. With the development of newer, more precise measuring technologies, shell strength can now be assessed in a consistent, objective fashion. A research project was conducted to determine what role shell strength might play in affecting external Salmonella Enteritidis contamination of egg contents. Visibly clean eggs were collected from an in-line shell egg-processing facility at the accumulator. Eggs were inoculated by dipping in a concentrated suspension of nalidixic acid-resistant Salmonella Enteritidis. After storage, eggs were assessed for shell strength and both external and internal Salmonella Enteritidis contamination. In the first study, there was a significant difference (P < 0.05) in shell strength among the three replicates. No differences between treatments were found for shell strength or Salmonella Enteritidis contamination of contents. In the second study, there were no replicate differences for any of the monitored factors. When rinsate and content samples were enriched, 100% of the rinsates were positive for Salmonella Enteritidis. No content samples were shown to be contaminated with Salmonella Enteritidis during direct plating, but 3 to 5% of the samples from each replicate were positive after enrichment. Correlation analysis of the results from each study found only weak correlations between shell strength and Salmonella Enteritidis contamination on eggshell surface or contents. Within the range of shell strengths recorded in this study, the correlation analysis suggests that shell strength does not play a major role in Salmonella Enteritidis contamination. Further work with eggs that represent a greater range of shell strengths could provide a clearer indication of the interaction of shell strength and Salmonella Enteritidis contamination. PMID- 16245703 TI - Antimicrobial susceptibilities of Salmonella isolates obtained from layer chicken houses on a commercial egg-producing farm in Japan, 1997 to 2002. AB - Susceptibility to antimicrobial agents was examined for 325 isolates of Salmonella enterica serotypes Cerro, Infantis, Livingstone, and Montevideo isolated from layer houses on a commercial egg-production farm in the western region of Japan between 1997 and 2002. No antimicrobials were used for therapeutic purposes on the farm during this period. From 1.8 to 3.1% of the isolates were resistant to ampicillin, chloramphenicol, and tetracycline. Resistance to streptomycin and sulfisoxazole was found in 52.9 and 65.5%, respectively, of Salmonella Montevideo isolates and in 0 to 13.2% of the isolates of the other serotypes. All the streptomycin-resistant isolates of Salmonella Montevideo also exhibited resistance to sulfisoxazole. Salmonella Montevideo isolates were first isolated in 1998, and 80.0% of the isolates obtained in this year were resistant to streptomycin and sulfisoxazole. The results suggest that Salmonella Montevideo isolates that had already acquired resistance may have been introduced into the layer houses, although the route and vehicle of transmission were uncertain. The proportion of Salmonella Montevideo isolates resistant to streptomycin and sulfisoxazole significantly decreased (P < 0.01) from 79.5% for 1998 through 1999 to 37.3% for 2000 through 2002. This decrease probably was due to the fact that no antimicrobials were used on the farm. Among 10 isolates from different serotypes obtained from 1997 to 2002 that were resistant to ampicillin, two and five isolates harbored 42- and 63-kb R plasmids and identical DraI restriction enzyme digest patterns, respectively, and carried the blaTEM gene. The results suggest that the ampicillin resistance determinants were transferred among different serotypes of Salmonella in the layer houses. PMID- 16245705 TI - Survival of Salmonella and Escherichia coli O157:H7 in chorizos. AB - Mexican-style raw meat sausages (chorizos) are not regulated in California when they are produced in small ethnic food markets. These sausages are sold uncooked, but their formulation imparts a color that may lead the consumer to assume that they are already cooked, and thus the chorizos may sometimes be eaten without proper cooking. If pathogens are present in such cases, illness may result. Survival of Salmonella and Escherichia coli O157:H7 in chorizos was evaluated under different storage conditions selected based on an initial survey of uninspected chorizos in California. Chorizos were formulated with five different initial water activity (aw) values (0.85, 0.90, 0.93, 0.95, and 0.97), stored under four conditions (refrigeration at 6 to 8 degrees C, room temperature at 24 to 26 degrees C, under a hood at 24 to 26 degrees C with forced air circulation, and incubation at 30 to 31 degrees C with convective air circulation), and sampled after 1, 2, 4, and 7 days. The initial pH was 4.8 and remained near 5.0 from day 1 of the sampling period. Two separate studies of packs inoculated with five-strain cocktails of Salmonella and of E. coli O157:H7 were performed twice for each initial aw. The three lowest aw values (0.85, 0.90, and 0.93) and the incubation and hood storage conditions were more effective (P < or = 0.05) at reducing the target pathogen levels in chorizos than were the two highest aw values (0.95 and 0.97) and the refrigeration storage condition, regardless of storage time. These results provide a scientific basis for guidelines given to producers of uninspected chorizo and should reduce the probability of foodborne illness associated with these products. PMID- 16245706 TI - Effect of acid shock with hydrochloric, citric, and lactic acids on the survival and growth of Salmonella typhi and Salmonella typhimurium in acidified media. AB - The effect of acid shock with hydrochloric, citric, or lactic acid on the survival and growth of Salmonella Typhi and Salmonella Typhimurium in acidified broth was evaluated. Salmonella serovars were acid shocked (1 h at 35 degrees C) in Trypticase soy broth acidified with hydrochloric, citric, or lactic acid at pH 5.5. Unshocked cells were exposed to the same media that had been neutralized before use to pH 7.0. Shocked and unshocked cells were inoculated into broth acidified with hydrochloric acid (pH 3.0), citric acid (pH 3.0), or lactic acid (pH 3.8), and growth and survival ability were evaluated. The acid shock conferred protection to Salmonella against the lethal effects of low pH and organic acids. The adaptive response was not specific to the anion used for adaptation. The biggest difference in reduction of survival between shocked and unshocked strains (approximately 2 log CFU/ml) was observed when the microorganisms were shocked with lactic acid and then challenged with citric acid. Salmonella Typhi was more tolerant of citric acid than was Salmonella Typhimurium, but Salmonella Typhimurium had higher acid tolerance in response to acid shock than did Salmonella Typhi. The acid shock decreased the extension of the lag phase and enhanced the physiological state values of Salmonella Typhi and Salmonella Typhimurium when the pH of growth was 4.5. This increased ability to tolerate acidity may have an important impact on food safety, especially in the case of Salmonella Typhi, given the very low infectious dose of this pathogen. PMID- 16245707 TI - Antimicrobial effect of Thai spices against Listeria monocytogenes and Salmonella typhimurium DT104. AB - The objective of this study was to determine the potential antimicrobial activity of extracts and essential oils of spices from Thailand against foodborne pathogenic bacteria. The antimicrobial efficacy of ginger (Zingiber officinale), fingerroot (Boesenbergia pandurata), and turmeric (Curcuma longa) was evaluated against five strains of Listeria monocytogenes and four strains of Salmonella enterica ssp. enterica serovar Typhimurium DT104. Antimicrobial activity was investigated in microbiological media by using an agar dilution assay and enumeration over time and a model food system, apple juice, by monitoring growth over time. In the agar dilution assay, water extracts of the three spices had no effect on L. monocytogenes. Similarly, 50% ethanol extracts of ginger or turmeric had no effect. In contrast, ethanolic fingerroot extracts at 5 to 10% (vol/ vol) inhibited most L. monocytogenes strains for 24 h in the agar dilution assay. Commercial essential oils (EO) of ginger or turmeric inhibited all L. monocytogenes at < or = 0.6 or < or = 10%, respectively. Fingerroot EO inhibited all strains at < or = 0.4%. In the enumeration-over-time assay, a 5% fingerroot ethanol extract reduced ca. 4 log CFU/ml Listeria by around 2 log in 24 h while 10% inactivated the microorganism in 9 h. Fingerroot EO at 0.2% inactivated 4 log CFU/ml L. monocytogenes in 6 to 9 h. Neither extracts nor commercial EO had any effect on Salmonella Typhimurium DT 104 with the exception of fingerroot EO, which inhibited all strains at < or = 0.7%. Addition of 0.2% fingerroot EO to apple juice reduced 4 log of L. monocytogenes Scott A and both strains of Salmonella Typhimurium to an undetectable level within 1 to 2 days. It was concluded that fingerroot EO and extract have potential for inhibiting pathogens in food systems. PMID- 16245708 TI - Evaluation of small-scale hot-water postpackaging pasteurization treatments for destruction of Listeria monocytogenes on ready-to-eat beef snack sticks and natural-casing wieners. AB - This study was conducted to evaluate small-scale hot-water postpackaging pasteurization (PPP) as a postlethality (post-cooking) treatment for Listeria monocytogenes on ready-to-eat beef snack sticks and natural-casing wieners. Using a commercially available plastic packaging film specifically designed for PPP applications and 2.8 liters of boiling water (100 degrees C) in a sauce pan on a hot plate, an average reduction in L. monocytogenes numbers of > or = 2 log units was obtained using heating times of 1.0 min for individually packaged beef snack sticks (three brands) and 4.0 min for packages of four sticks (two brands) and seven sticks (three brands). Average product surface temperatures, measured as soon as possible after PPP and opening the package, were 47 to 51.5, 58 to 61.5, and 58.5 to 61 degrees C for the beef snack sticks packages of one, four, and seven sticks per package, respectively. A treatment of 7.0 min for packages of four natural-casing wieners (three brands) achieved L. monocytogenes reductions of > or = 1.0 log unit and average product surface temperature of 60.5 to 63.5 degrees C. Cooked-out fat and moisture resulting from tested treatments ranged from 0.2 to 1.1% by weight for beef snack sticks and from 0.4 to 1.2% by weight for natural-casing wieners. For natural-casing wieners, PPP had no detrimental effect on overall product desirability to consumers; results suggested that PPP may significantly enhance appearance of this product. However, for beef snack sticks the cooking out of fat and moisture during PPP had a significant negative effect on consumer opinions of product appearance. PMID- 16245709 TI - Antilisterial activity of a Carnobacterium piscicola isolated from Brazilian smoked fish (surubim [Pseudoplatystoma sp.]) and its activity against a persistent strain of Listeria monocytogenes isolated from surubim. AB - Data on the prevalence and growth of Listeria monocytogenes in lightly preserved fish products from subtropical and tropical regions are very scarce. Our research describes L. monocytogenes that was detected in 5% of the packages of cold-smoked surubim, a native Brazilian freshwater fish that we analyzed, and shows that the strains isolated were of the same random amplified polymorphic DNA subtype as the strains that were isolated from the same factory 4 years earlier. A bacteriocinogenic strain of Carnobacterium piscicola (strain C2), isolated from vacuum-packed cold-smoked surubim, and two C. piscicola strains, isolated from vacuum-packed, cold-smoked salmon, were capable of limiting or completely inhibiting the growth of an L. monocytogenes (strain V2) isolated from surubim in fish peptone model systems incubated at 10 degrees C. Monocultures of L. monocytogenes reached 108 CFU/ml (g), whereas the growth of L. monocytogenes was completely inhibited by C. piscicola C2. The bacteriocinogenic C. piscicola A9b+ and its nonbacteriocinogenic mutant A9b- reduced maximum Listeria levels by 2 to 3 log units. Both bacteriocinogenic C. piscicola strains prevented listerial growth in cold-smoked fish juices (surubim and salmon). Although the carnobacteria grew poorly on cold-smoked surubim at 10 degrees C, the strains were able to reduce maximum Listeria counts by 1 to 3 log units in an artificially inoculated product (surubim). We conclude that Brazilian smoked fish products harbor L. monocytogenes and should be stabilized against the growth of the organism. C. piscicola C2 has the potential for use as a bioprotective culture in surubim and other lightly preserved fish, but further studies are required to optimize its effect. PMID- 16245710 TI - Interactions of meat-associated bacteriocin-producing Lactobacilli with Listeria innocua under stringent sausage fermentation conditions. AB - The kinetics of the antilisterial effect of meat-associated lactobacilli on Listeria innocua LMG 13568 were investigated during laboratory batch fermentations. During these fermentations, which were performed in a liquid meat simulation medium, a combination of process factors typical for European-style sausage fermentations was applied, such as a temperature of 20 degrees C and a representative pH and salting profile. Two bacteriocin-producing sausage isolates (Lactobacillus sakei CTC 494 and Lactobacillus curvatus LTH 1174), which have already proven efficacy in sausage trials, and one nonbacteriocinogenic, industrial strain (Lactobacillus sakei I), were evaluated. Staphylococcus carnosus 833 was included in the experiment because of its role in flavor and color development. When grown as a monoculture or upon cocultivation with L. sakei I and S. carnosus 833, L. innocua LMG 13568 developed slightly, despite the stress of low temperature, pH, lactic acid, salt, and nitrite. In contrast, when either of the bacteriocin producers was used, the L. innocua LMG 13568 population was rapidly inactivated with more than 3 log CFU ml(-1) after 2 days of fermentation. A bacteriocin-tolerant L. innocua LMG 13568 subpopulation (4 X 10( 4)) remained after bacteriocin inactivation. Thus, when the initial level of L. innocua LMG 13568 equaled 3 log CFU ml(-1), all cells were inactivated and no bacteriocin-tolerant cells were detected, even after 7 days of incubation. S. carnosus was not inactivated by the Lactobacillus bacteriocins and displayed slight growth. PMID- 16245711 TI - Stability of enterocin AS-48 in fruit and vegetable juices. AB - Enterocin AS-48 is a candidate bacteriocin for food biopreservation. Before addressing application of AS-48 to vegetable-based foods, the interaction between AS-48 and vegetable food components and the stability of AS-48 were studied. Enterocin AS-48 had variable interactions with fruit and vegetable juices, with complete, partial, or negligible loss of activity. For some juices, loss of activity was ameliorated by increasing the bacteriocin concentration, diluting the juice, or applying a heat pretreatment. In juices obtained from cabbage, cauliflower, lettuce, green beans, celery, and avocado, AS-48 was very stable for the first 24 to 48 h of storage under refrigeration, and decay of activity was markedly influenced by storage temperature. In fresh-made fruit juices (orange, apple, grapefruit, pear, pineapple, and kiwi) and juice mixtures, AS-48 was very stable for at least 15 days at 4 degrees C, and bacteriocin activity was still detectable after 30 days of storage. Gradual and variable loss of activity occurred in juices stored at 15 and 28 degrees C; inactivation was faster at higher temperatures. In commercial fruit juices (orange, apple, peach, and pineapple) stored at 4 degrees C, the bacteriocin was completely stable for up to 120 days, and over 60% of initial activity was still present in juices stored at 15 degrees C for the same period. Commercial fruit juices stored at 28 degrees C for 120 days retained between 31.5% (apple) and 67.71% (peach) of their initial bacteriocin activity. Solutions of AS-48 in sterile distilled water were stable (120 days at 4 to 28 degrees C). Limited loss of activity was observed after mixing AS-48 with some food-grade dyes and thickening agents. Enterocin AS-48 added to lettuce juice incubated at 15 degrees C reduced viable counts of Listeria monocytogenes CECT 4032 and Bacillus cereus LWL1 to below detection limits and markedly reduced viable counts of Staphylococcus aureus CECT 976. PMID- 16245712 TI - Reduction of fumonisin B1 and zearalenone by lactic acid bacteria in fermented maize meal. AB - Fusarium species are fungi that infect maize products worldwide and elaborate mycotoxins, which have been associated with cancer. This study was carried out to investigate the potential of lactic acid bacteria fermentation in reducing mycotoxin concentration and toxicity in maize meal products. Maize meal was spiked separately with fumonisin B1 and zearalenone and then allowed to ferment for 4 days. The potential cytotoxicity of the mycotoxin-spiked fermented extracts was also investigated using the SNO human esophageal carcinoma cell line (the SNO cell line was explanted from a cancer patient, S.N., a 62-year-old Zulu man, in July 1972). A significant decrease (P < 0.05) in the concentration of the two mycotoxins was observed, with a 56 to 67% and a 68 to 75% reduction in the third and fourth days, respectively. The two mycotoxins were not detectable in commercially fermented maize meal (amahewu) samples. After fermentation, mycotoxin-spiked maize meal samples containing lactic acid bacteria culture were comparatively less toxic to SNO cells than were samples without lactic acid bacteria. However, this difference in toxicity was not significant (P > 0.05). These results indicate that lactic acid bacteria fermentation can significantly reduce the concentration of mycotoxins in maize. However, such a reduction may not significantly alter the possible toxic effects of such toxins. The exact mechanism of toxin reduction warrants further investigation. PMID- 16245713 TI - Integration of biocontrol agents and food-grade additives for enhancing protection of stored apples from Penicillium expansum. AB - Forty-nine compounds currently used as additives in foods were tested in combination with three biocontrol agents, the yeasts Rhodotorula glutinis, Cryptococcus laurentii, and the yeastlike fungus Aureobasidium pullulans, to increase their antagonistic activity against Penicillium expansum, the causal agent of blue mold on apples. Twelve additives dramatically improved the antagonistic activity of one or more of the tested biocontrol agents. In a two way factorial experiment with these selected additives the percentage of P. expansum rots on apples was significantly influenced by the antagonist and the additive as well as by their interaction. The combination of the biocontrol agents and some additives resulted in a significantly higher activity with respect to the single treatments applied separately, producing additive or synergistic effects. Some of the selected additives combined with a low yeast concentration (106 cells per ml) had comparable or higher efficacy than the biocontrol agents applied alone at a 100-fold higher concentration (10(8) cells per ml). Some organic and inorganic calcium salts, natural gums, and some antioxidants displayed the best results. In general, the effect of each additive was specific to the biocontrol isolate used in the experiments. Possible mechanisms involved in the activity of these beneficial additives and their potential application in effective formulations of postharvest biofungicides are discussed. PMID- 16245714 TI - Fate of ochratoxin a during cooking of naturally contaminated polished rice. AB - Ochratoxin A (OTA), a mycotoxin widespread in cereals, occurs in polished rice that is consumed as cooked rice after washing and steaming. Cooking decreases OTA levels in food to varying extents, but little is known about how cooking changes the biological activity of this mycotoxin. We therefore evaluated the fate of OTA during rice cooking to determine the OTA residues and cytotoxic potential in vitro. Water-washed rice, ordinary cooked rice, and pressure-cooked rice were prepared from three polished rice lots naturally contaminated with OTA. Residual OTA in each sample was analyzed by high-performance liquid chromatography (HPLC), whereas in vitro cytotoxicity of OTA to C6 glioma cells, susceptible to low levels (nanograms per milliliter) of OTA, was used to confirm the chemical analysis. OTA concentration, as determined by HPLC analysis, in the cooked rice by both types of cookers was significantly lower than (59 to 75%) in the raw polished rice and water-washed rice. The cytotoxicity of the OTA that remained in the pressure-cooked rice from three lots was markedly decreased (approximately 20%, P < 0.05) when compared with other samples in respective lots. This confirms that cooking lowers OTA residues. Although washing polished rice with water had little effect on OTA levels, pressure steaming appeared to be the critical cooking step not only to reduce OTA residues in polished rice before reaching the consumer as the dietary staple of cooked rice, but also to diminish cytotoxicity of OTA. PMID- 16245715 TI - Variability of virus attachment patterns to butterhead lettuce. AB - Enteric viruses account for most foodborne illness in the United States. The objective of this study was to determine whether the isoelectric point (pI) of viruses such as feline calicivirus (FCV), echovirus 11, and bacteriophages phiX174 and MS2 had any effect on their attachment to butterhead lettuce. The adsorption of virus particles to the lettuce was variable. Bacteriophage MS2 was the only virus that fit the current Derjaguin-Landau-Verway-Overbeek model of virus attachment. Echovirus 11 had the highest affinity to lettuce surface. Echovirus 11 appeared to exhibit reversible attachment above its pI, whereas below its pI strong adsorption was observed. Adsorption of FCV was at its maximum above its pI. Bacteriophage phiX174 exhibited the most complex adsorption pattern, with attachment occurring only at the pH extremes (pH 3.0 and 8.0). These results suggest the current model for virus adsorption to sediment does not adequately explain the attachment of virus to lettuce. Importantly, the results indirectly suggest that current sample processing methods to recover viruses from lettuce may differentially select for the recovery of only certain virus types. PMID- 16245716 TI - Effects of temperature and moisture on coliphage PRD-1 survival in soil. AB - The goal of this study was to quantitatively assess the effects of temperature and soil moisture on the survival of coliphage PRD-1 in soil. PRD-1 was added to sandy loam soil at five different soil moisture levels. The soil seeded with PRD 1 was packed into sterile polyethylene jars and exposed to eight different temperatures in an oven. Samples were collected over 14 to 25 days depending on the temperature. The inactivation rate of PRD-1 increased linearly with increased temperature. The inactivation rate gradually decreased when the soil moisture level decreased from 20.9 to 8.9%. However, the inactivation rate increased when the soil moisture content reached 5.1%, suggesting the existence of an optimal soil moisture condition for PRD-1 survival. It is also possible that there is a threshold soil moisture level below which the inactivation of PRD-1 suddenly increases. Marked reductions in recoveries were observed as the soil moisture approached or fell below 5.0% as a result of evaporation. The increased inactivation of PRD-1 due to strong association with soil particles may have caused rapid reductions in recoveries. The evaporation process appeared to affect PRD-1 survival substantially at higher temperatures whereas little effect was observed at lower temperatures. A model developed from this study predicted PRD-1 survival in subsurface soil in field conditions with an average error of 11.0%. PMID- 16245717 TI - Establishment of a novel multiplex PCR assay and detection of toxigenic strains of the species in the Bacillus cereus group. AB - Five different enterotoxins and one emetic toxin of Bacillus cereus have been characterized. To amplify all of the enterotoxin and emetic-specific sequences of the species in the B. cereus group, a multiplex PCR with 12 primer pairs was established. In developing the assay method, a common terminal sequence at the 3' ends of all primers was chosen and a hot start Taq polymerase was used to overcome primer dimer formation. The assay was successfully applied to analyze the toxigenic potential of 162 food-poisoning and food-related strains. Results showed that there were 10 toxigenic patterns for all the test strains. All of the B. cereus strains carried at least one toxin gene. More than 70% of Bacillus mycoides strains carried no known toxin genes. The toxin profiles and toxin genes of B. mycoides strains were significantly different from B. cereus strains (P < 0.05), although the two species were closely related. The results suggest that many B. mycoides strains might be less prone to cause food poisoning. They also indicate the importance of detecting the toxin genes together with the detection of the species in the B. cereus group. PMID- 16245718 TI - A robotic DNA purification protocol and real-time PCR for the detection of Campylobacter jejuni in foods. AB - Primers designed to amplify a Campylobacter jejuni cadF gene sequence were used in an SYBR Green I real-time PCR assay as an alternative to conventional bacteriological methods for the rapid detection of C. jejuni in foods. Twenty five grams of chicken skin (breast and thigh) was contaminated by adding approximately 1, 10, or 50 CFU of C. jejuni ATCC 35560. Twenty-five grams of pork and 25-ml aliquots of milk were also inoculated with 1 and 10 CFU of the pathogen. The samples were incubated in Bolton broth for different periods at 37 and 42 degrees C under microaerophilic conditions. Using a commercial robotic DNA purification system, DNA was extracted and purified from 1-ml aliquots of the enrichment cultures before and after centrifugation of the 250-ml enrichment broth at 15,900 x g for 10 min at 4 degrees C. The DNA was used as the template in a real-time PCR assay. C. jejuni was detected after 12 h of enrichment from samples inoculated with about 50 CFU/25-g sample. After centrifugation, an enrichment step of 8 h was sufficient to allow detection of pathogen in samples inoculated with 10 CFU/25 g. However, 24 h of enrichment was necessary to detect pathogen in samples inoculated with approximately 1 CFU/25 g. The real-time PCR protocol developed in this study significantly reduced the detection time of C. jejuni in foods. PMID- 16245719 TI - Multiplex-touchdown PCR assay for the detection and genotyping of Helicobacter pylori from artificially contaminated sheep milk. AB - Helicobacter pylori (Hp) is an organism commonly present worldwide in the human population, sometimes causing serious illnesses such as duodenal and gastric ulcers, adenocarcinoma of the stomach, and low-grade B-cell mucosa-associated lymphoid tissue lymphoma of the stomach. This article describes a multiplex touchdown PCR method for the identification and genotyping (vacA-s1/m1, sl/m2, and s2/m2-and cagA genes) of Hp directly from sheep milk artificially contaminated with Hp strains from human gastric biopsies and with Hp ATCC 43504. The strains from humans carried sl/m2 cagA+ and s2/m2 cagA allelic combinations, while the ATCC strains carried an sl/ml cagA+ allelic combination. The technique showed a sensitivity of 15 CFU/ml for species identification and of 1,500 CFU/ml for the detection of genes encoding for VacA and CagA. It has proven to be specific and rapid, and the authors suggest that it be used as a rapid screening method to ensure that sheep milk is uncontaminated with this organism. PMID- 16245720 TI - Development of a rapid immunochromatographic strip for detection of Escherichia coli O157. AB - We developed an immunochromatographic (IC) strip for the rapid detection of Escherichia coli O157 in enriched samples. Murine monoclonal antibody to E. coli O157:H7 lipopolysaccharide was conjugated with 40 nm of colloidal gold particles by the citrate method. The specificity of the IC strip was determined using 48 pure-cultured bacteria, including 32 E. coli strains and 16 non-E. coli strains. Regardless of H serotype, E. coli O157 strains produced a positive signal, whereas the others, representing 29 E. coli serotypes, did not. Among 16 non-E. coli strains, only Citrobacter amalonaticus yielded a positive signal. The sensitivity of the IC strip was determined using 10-fold diluted E. coli O157:H7, with a range of 1.8 X 10(7) to 1.8 CFU/ml in enriched raw beef. E. coli O157 could be detected at a minimum of 1.8 x 10(5) CFU/ml without enrichment and 1.8 CFU/ml after enrichment. Various samples were enriched to detect E. coli O157 using the IC strip and to isolate E. coli O157:H7 using traditional culture procedures. The IC strip test results exhibited 100% agreement with traditional methods after selective enrichment, since E. coli O157:H7 was also isolated from all the samples with positive strip test results. However, the specificity of the strip was somewhat higher with pork (98.8%) than with bovine feces (87.9%) and swine feces (93.4%). These results indicated that the IC strip exhibits high specificity and sensitivity in the detection of E. coli O157, and this assay is rapid, economical, and simple, without requirement of complicated equipment. PMID- 16245721 TI - Shell rinse and shell crush methods for the recovery of aerobic microorganisms and enterobacteriaceae from shell eggs. AB - Recovery of bacteria from shell eggs is important for evaluating the efficacy of processing and the quality and safety of the final product. Shell rinse (SR) techniques are easy to perform and widely used. An alternative sampling method involves crushing and rubbing the shell (CR). To determine the most appropriate method for recovering microorganisms from shell eggs, 358 shell eggs were collected from a commercial egg processor and sampled by SR and CR techniques. Total aerobic mesophiles and Enterobacteriaceae were enumerated on plate count and violet red bile glucose agar plates, respectively. Unwashed, in process, and postprocess eggs were evaluated in the study. Aerobic microorganism prevalence for eggshells sampled was similar for both methods (approximately 100%), but the log CFU per milliliter values were higher in the SR than the CR samples (3.2 and 2.2, respectively). Average Enterobacteriaceae recovery was similar for both methods (45 versus 40% for the SR and CR methods, respectively) when all eggs were considered together. This population was detected more often by SR when unwashed eggs were sampled (90 versus 56% for the SR and CR methods, respectively), equally by SR and CR for in-process eggs (30 versus 29.3% for the SR and CR methods, respectively), but more often by CR for postprocess eggs (10 versus 36% for the SR and CR methods, respectively). The SR technique was easier to perform and recovered larger numbers of aerobic organisms, particularly for unwashed eggs. However, the CR technique was more efficient for recovery of Enterobacteriaceae from postprocess eggs. Stage of shell egg processing may be an important consideration when choosing egg sampling methods. PMID- 16245722 TI - Effect of temperature, pH, and water activity on biofilm formation by Salmonella enterica enteritidis PT4 on stainless steel surfaces as indicated by the bead vortexing method and conductance measurements. AB - An assay was developed in an effort to elucidate the effect of important environmental parameters (temperature, pH, and water activity [aw]) on Salmonella Enteritidis biofilm formation on stainless steel surfaces. To achieve this, a modified microbiological technique used for biofilm studying (the bead vortexing method) and a rapid method based on conductivity measurements were used. The ability of the microorganism to generate biofilm on the stainless surfaces was studied at three temperatures (5, 20, and 37 degrees C), four pH values (4.5, 5.5, 6.5, and 7.4), and four aw values (0.5, 1.5, 5.5, and 10.5% NaCl). Results obtained by the bead vortexing method show that maximum numbers of adherent bacteria per square centimeter (106 CFU/cm2) were attained in 6 days at 20 degrees C. Biofilm formation after 7 days of incubation at 20 degrees C was found to be independent of the pH value. In addition, the high concentration of sodium chloride (10.5% NaCl, aw = 0.94) clearly inhibited the adherence of cells to the coupons. Conductance measurements were used as a supplementary tool to measure indirectly the attachment and biofilm formation of bacterial cells on stainless steel surfaces via their metabolic activity (i.e., changes in the conductance of the growth medium due to microbial growth or metabolism). Results obtained by conductance measurements corresponded well to those of the bead vortexing method. Furthermore, we were able to detect cells that remained attached on the metal surfaces even after vortexing via their metabolic activity. The results, except for demonstrating environmental-dependent Salmonella Enteritidis biofilm formation, indicated that traditional vortexing with beads did not remove completely biofilm cells from stainless steel; hence, conductance measurements seem to provide a more sensitive test capable to detect down to one single viable organism. PMID- 16245723 TI - A comparison of wet-dry swabbing and excision sampling methods for microbiological testing of bovine, porcine, and ovine carcasses at red meat slaughterhouses. AB - A comparison of wet-dry swabbing and surface tissue excision of carcasses by coring was undertaken. Samples from 1,352 bovine, 188 ovine, and 176 porcine carcasses were collected from 70 separate visits to commercial slaughterhouses operating under normal conditions. The mean total aerobic viable bacterial counts (TVCs) for all species sampled by excision was 5.36 log units, which was significantly greater than the 4.35 log units measured for swabbing. Poorly correlated linear relationships between swab- and excision-derived bacterial numbers from near-adjacent carcasses were observed for all three animal species. R2 values for least squares regressions for bovine, ovine, and porcine carcasses were 0.09, 0.27, and 0.21, respectively. The reasons why it was not possible to calculate a factor that allowed the interconversion of bacterial numbers between samples collected by each sampling method were investigated. Uncertainty associated with laboratory analyses was a contributing factor because the geometric relative standard deviations measured for TVCs were 0.174 and 0.414 for excision and swabbing, respectively. Uneven distribution of bacteria at identical sampling sites on near-adjacent carcasses on processing lines was also a contributory factor. The implications of these findings for process control verification were investigated by intensive sampling for 13 weeks in three commercial slaughterhouses. As many as 4 log units of difference in TVCs were observed in duplicate samples collected within a narrow timeframe from near adjacent carcasses on the processing line. We conclude that it might not be appropriate to institute corrective actions in slaughterhouses on the basis of a single week's test results. PMID- 16245724 TI - Experimental comparison of excision and swabbing microbiological sampling methods for carcasses. AB - Bovine sides, ovine carcasses, and porcine carcasses were individually inoculated by dipping in various suspensions of a marker organism (Escherichia coli K-12 or Pseudomonas fluorescens), alone or in combination with two meat-derived bacterial strains, and were sampled by two standard methods: cotton wet-dry swabbing and excision. The samples were examined for bacterial counts on plate count agar (PCA plate counts) and on violet red brilliant green agar (VRBGA plate counts) by standard International Organization for Standardization methods. Average bacterial recoveries by swabbing, expressed as a percentage of the appropriate recoveries achieved by excision, varied widely (2 to 100%). Several factors that potentially contributed to relatively low and highly variable bacterial recoveries obtained by swabbing were investigated in separate experiments. Neither the difference in size of the swabbed area (10, 50, or 100 cm2 on beef carcasses) nor the difference in time of swabbing (20 or 60 min after inoculation of pig carcasses) had a significant effect on the swabbing recoveries of the marker organism used. In an experiment with swabs preinoculated with the marker organism and then used for carcass swabbing, on average, 12% of total bacterial load was transferred inversely (i.e., from the swab to the carcass during the standard swabbing procedure). In another experiment, on average, 14% of total bacterial load was not released from the swab into the diluent during standard swab homogenization. Use of custom-made swabs with abrasive butts, around which metal pieces of pan scourers were wound, markedly increased PCA plate count recoveries from noninoculated lamb carcasses at commercial abattoirs compared with cotton swabs. In spite of the observed inferiority of the cotton wet-dry swabbing method compared with the excision method for bacterial recovery, the former is clearly preferred by the meat industry because it does not damage the carcass. Therefore, further large-scale evaluation of the two carcass sampling methods has been undertaken under commercial conditions and reported separately. PMID- 16245725 TI - In situ sample cleanup during immunoassay: a simple method for rapid detection of aflatoxin B, in food samples. AB - A strategy for rapid in situ elimination of interfering substances that are present in extracts of food samples during assay is described in this article. The novel feature of this method is that the sample purification is carried out as a part of the assay, and a separate sample cleanup step is not required. The assay procedure involves the sequential addition of standard or sample, cleaning solutions, and aflatoxin B1-horseradish peroxidase conjugate (AFB1-HRP) over antibody-spotted zones of a membrane, and 3,3'-diaminobenzidine was used as the substrate for visualization. We have determined that trifluoroacetic acid and propionic acids at concentrations of 100 mM are highly effective for cleaning groundnut, wheat, corn, and poultry feed samples and that NaHCO3 (100 mM) is successful in cleaning processed soybean. In all cases, subsequent washing was performed with phosphate-buffered saline solution to facilitate the removal of traces of adhering interfering substances. A batch of 12 samples can be analyzed within 8 min either by visual comparison of the color intensity (inversely related to the analyte concentration) of a sample spot with those of reference standards or, more precisely, by densitometry. The method was tested for the analysis of AFB1 in groundnut, wheat, corn, processed soybean, chili, and poultry feed. The detection limit obtained was 5 microg/kg, except for chili, where it was 10 microg/kg. The average recoveries from different noninfected food samples spiked with AFB1 at concentrations of 5 to 100 microg/kg were between 99 and 105%. The values obtained for infected corn and groundnut samples correlated well with the estimates obtained by high-pressure liquid chromatography. The absence of a sample extraction step reduces the cost and labor involved in the assay. The method may be potentially applicable to the assay of other mycotoxins and environmental pollutants. PMID- 16245726 TI - Detection of bovine central nervous system tissue in liver sausages using a reverse transcriptase PCR technique and a commercial enzyme-linked immunosorbent assay. AB - The suitability of a reverse transcriptase (RT) PCR assay was evaluated for the detection of bovine central nervous system (CNS) tissue specifically in liver sausages. Because of its emulsifying effect, CNS tissue was frequently added to this kind of meat product in the past. On standard samples, the RT-PCR technique reliably detected a concentration of 0.25% bovine CNS tissue in liver sausages stored for up to 28 days. Following the successful application of RT-PCR for the detection of bovine CNS tissue in these specially prepared samples, a field study was performed with a total of 258 liver sausages purchased in retail markets. All sausages were tested with both an RT-PCR assay and a commercial enzyme-linked immunosorbent assay (ELISA) kit. Nine (3.5%) of the retail liver sausage samples were positive for CNS tissue in the ELISA, but none were positive for this tissue in the RT-PCR assay. All positive ELISA results indicated the presence of 0.23 to 0.30% CNS tissue. Recent studies have indicated that the RT-PCR assay is not as sensitive for porcine CNS tissue as for bovine CNS tissue, which this assay can detect at 0.25%. Although the ELISA is not species specific, the CNS tissue detected by the ELISA is assumed to stem from a nonbovine species. The RT-PCR technique is a sensitive tool for the detection of bovine CNS tissues in a problematic matrix such as liver sausage. ELISA screening followed by a species specific RT-PCR assay for bovine CNS tissue is a practical approach for monitoring meat products for compliance with European food regulations. PMID- 16245727 TI - Beliefs about meals eaten outside the home as sources of gastrointestinal illness. AB - In a 2002 telephone survey of 16,435 randomly selected U.S. residents, respondents answered several questions about their beliefs concerning sources of gastrointestinal illness. Of those who had experienced vomiting or diarrhea in the month before their telephone interview, 22% believed the source of their gastrointestinal illness was a meal eaten outside the home. I11 respondents who had diarrhea but not vomiting and who did not miss work because of their illness were more likely to believe the illness resulted from a specific outside meal. I11 respondents attributed their illness to a specific outside meal for several reasons, including symptom timing (43%) and illness of their meal companions (6%). Eight percent of ill respondents reported their illness to a health department or the restaurant suspected of causing the illness. Those with vomiting and those who missed work or activities because of their illness were more likely to report their illness. Most respondents (54%) who attributed their illness to a specific outside meal said their illness symptoms began within a short time (5 h) of eating that meal. The foodborne illnesses for which this is a likely time frame typically are associated with vomiting, but respondents with vomiting did not report a shorter symptom onset than respondents without vomiting. These findings suggest that ill respondents may have the misconception that foodborne illness symptoms typically occur shortly after ingestion of contaminated food. Results suggest that education efforts should focus on the nature and timing of foodborne illness symptoms and the importance of reporting suspected foodborne illnesses. PMID- 16245728 TI - Dietary intake of polycyclic aromatic hydrocarbons in a Spanish population. AB - The objective of the present study was to estimate the dietary intake of polycyclic aromatic hydrocarbons (PAHs), particularly benzo[a]pyrene (B[a]P), as well as to identify the principal dietary sources of such compounds in the Spanish adult population. The study included 40,690 subjects aged 35 to 64 years from five regions of Spain that were included in the European Prospective Investigation into Cancer and Nutrition (EPIC)-Spain cohort. Usual food intake was estimated by personal interview through a computerized version of a dietary history questionnaire. The estimations of B[a]P and total PAHs were made, taking into account the country where the determinations of content of these compounds in the foods came from and the year of publication. The mean intake of B[a]P in the population was 0.14 microg/day, and the mean intake of total PAHs was 8.57 microg/day. Both for B[a]P and total PAHs, women had a significantly lower mean intake than men, and older people consumed lesser amounts than younger people. Furthermore, the intake was higher in the northern regions. There were no significant differences by smoking status. The food groups of meat and meat products, cereals, and oils and fats contribute 55.5% to the total B[a]P intake, while cereals and meat and meat products contribute 61% to the total PAH consumption. Our estimations of B[a]P intake were lower than in the United Kingdom and The Netherlands, were similar to those found in other studies from Spain and Italy, and were higher than those in the United States and Norway. PMID- 16245729 TI - Recovery of presumptive Alicyclobacillus strains from orange fruit surfacest. AB - Objectives of this research were to investigate the detection frequency of presumptive Alicyclobacillus strains, also known as thermoacidophilic or acidothermophilic bacteria, on oranges entering juice-processing facilities and to compare results from three common isolation agars (acidified potato dextrose agar, Ali agar, and K agar). A total of 1,575 fruits were sampled from three points (ungraded fruits, graded sound fruits, and graded defective fruits) at two juice-processing facilities during two harvest seasons. Buffer used to rinse individual fruits was assayed for the presence of thermoacidophilic bacteria using an enrichment procedure. Isolates were considered presumptive Alicyclobacillus if they were gram-positive, sporogenous, rod-shaped bacteria, with growth at 45 degrees C and no or slight growth at 25 degrees C on a low pH medium (pH 3.7) coupled with lack of growth on a neutral pH medium (pH 7.0) at both temperatures and a lack of growth in Sulfobacillus broth medium (pH 2.0). More than one third of all fruits sampled at the two facilities were contaminated with presumptive Alicyclobacillus strains. Therefore, incoming fruits are a substantial means by which these organisms gain entrance to the processing facility. Significantly (P < or = 0.05) more detection was observed with a mineral-containing medium (Ali agar) than with nonmineral containing media (K agar and acidified potato dextrose agar). PMID- 16245730 TI - Shelf-life extension of refrigerated Mediterranean mullet (Mullus surmuletus) using modified atmosphere packaging. AB - The present work evaluated the quality and freshness characteristics and the effect of modified atmosphere packaging (MAP) on the shelf-life extension of refrigerated Mediterranean mullet using microbiological, biochemical, and sensory analyses. Fresh open sea red mullet (Mullus surmuletus) were packaged in four different atmospheres: M1, 10%/20%/70% (O2/ CO2/N2); M2, 10%/40%/50% (O2/CO2/N2); M3, 10%/60%/30% (O2/CO2/N2); identical fish samples were packaged in air. All fish were kept under refrigeration (4 +/- 0.5 degrees C) for 14 days. Of the three gas atmospheres, the 10%/40%/50% (M2) and 10%/ 60%/30% (M3) gas mixtures were the most effective for inhibiting growth of aerobic microflora in mullet samples until day 10 of refrigerated storage. H2S-producing bacteria and pseudomonads were part of the mullet microflora and their growth was partly inhibited under MAP conditions. Between these two bacterial groups, H2S-producing bacteria (including Shewanella putrefaciens) were dominant toward the end of the storage period, regardless of the packaging conditions. Brochothrix thermosphacta and lactic acid bacteria were found to be members of the final microbial flora of MAP and air-packaged mullet, whereas the Enterobacteriaceae population was lower than other bacterial groups. Of the chemical freshness indices determined, thiobarbituric acid values were variable in mullet samples irrespective of packaging conditions indicative of no specific oxidative rancidity trend. Based on sensorial data and aerobic plate count, trimethylamine nitrogen and total volatile basic nitrogen limit values in the range of ca. 15 to 23 and 52 to 60 mg N/100 g of fish muscle were obtained, respectively, for mullet packaged under modified atmosphere and air. Sensory analyses (odor and taste attributes) showed that the limit of sensorial acceptability was reached after ca. 6 days for the samples packaged in air, 8 days for the M1 and M3 samples, and after 10 days for the M2 samples. Respective shelf-life extension for fresh whole mullet was ca. 2 days (M1 and M3 gas mixtures), and 4 days (M2 gas mixture). PMID- 16245731 TI - Prevalence and characterization of typical and atypical Escherichia coli from fish sold at retail in Cochin, India. AB - Escherichia coli is a common contaminant of seafood in the tropics and is often encountered in high numbers. The count of E. coli as well as verotoxigenic E. coli O157:H7 was estimated in 414 finfish samples composed of 23 species of fresh fish from retail markets and frozen fish from cold storage outlets in and around Cochin, India. A total of 484 presumptive E. coli were isolated, and their indole methyl red-Voges-Proskauer-citrate (IMViC) pattern was determined. These strains were also tested for labile toxin production by a reverse passive latex agglutination method and checked for E. coli serotype O157 by latex agglutination with O157-specific antisera. Certain biochemical marker tests, such as methylumbelliferyl-beta-glucuronide (MUG), sorbitol fermentation, decarboxylase reactions, and hemolysis, which are useful for screening pathogenic E. coli, were also carried out. Results showed that 81.4% of the E. coli isolates were sorbitol positive. Among this group, 82% were MUG positive, and 14.46% of the total E. coli isolates showed human blood hemolysis. None of the isolates were positive for agglutination with E. coli O157 antisera nor did any produce heat-labile enterotoxin. This study indicates that typical E. coli O157 or labile toxin producing E. coli is absent in the fish and fishery environments of Cochin (India). However, the presence of MUG and sorbitol-negative strains that are also hemolytic indicates the existence of aberrant strains, which require further investigation. PMID- 16245732 TI - Efficacy of UV light for the reduction of Listeria monocytogenes in goat's milk. AB - Certain types of goat's cheeses are produced using unpasteurized milk, which increases the food safety concerns for these types of products. Popularity and consumption of goat's milk products have increased, and the niche market includes gourmet goat's cheeses. The U.S. Code of Federal Regulations and the Pasteurized Milk Ordinance both address the possibility for processing alternatives to heat treatment, and the use of UV light treatment may be a viable alternative that still ensures the safety of the product. Fresh goat's milk was inoculated with Listeria monocytogenes (L-2289) at 10(7) CFU/ml and exposed to UV light using the CiderSure 3500 apparatus (FPE Inc., Macedon, NY). Inoculated milk was exposed to a UV dose range between 0 and 20 mJ/cm2 to determine the optimal UV dose. A greater than 5-log reduction was achieved (P < 0.0001) when the milk received a cumulative UV dose of 15.8 +/- 1.6 mJ/cm2. The results of this study indicate that UV irradiation could be used for the reduction of L. monocytogenes in goat's milk. PMID- 16245733 TI - Concentrations of antibiotic residues vary between different edible muscle tissues in poultry. AB - Antibiotics are used by veterinarians and producers to treat disease and improve animal production. The federal government, to ensure the safety of the food supply, establishes antibiotic residue tolerances in edible animal tissues and determines the target tissues (e.g., muscle) for residue monitoring. However, when muscle is selected as the target tissue, the federal government does not specify which type of muscle tissue is used for monitoring (e.g., breast versus thigh). If specific muscle tissues incorporate residues at higher concentrations, these tissues should be selected for residue monitoring. To evaluate this possibility in poultry, chickens were divided into four groups and at 33 days of age were dosed with enrofloxacin (Baytril), as per label directions, at either 25 ppm for 3 days, 25 ppm for 7 days, 50 ppm for 3 days, or 50 ppm for 7 days. Breast and thigh muscle tissues were collected from each bird (n = 5 birds per day per group) during the dosing and withdrawal period, and fluoroquinolone concentrations were determined. The results indicate higher overall enrofloxacin concentrations in breast versus thigh muscle for each treatment group (P < 0.05). These data indicate, at least for enrofloxacin, that not all muscle tissues incorporate antibiotics at the same concentrations. These results may be helpful to regulatory agencies as they determine what tissues are to be monitored to ensure that the established residue safety tolerance levels are not exceeded. PMID- 16245734 TI - Reduction in flock prevalence of Campylobacter spp. in broilers in Norway after implementation of an action plan. AB - An action plan against thermophilic Campylobacter spp. in Norwegian broilers was implemented in May 2001. The action plan consists of three parts: a surveillance program including all Norwegian broiler flocks slaughtered before 50 days of age, a follow-up advisory service on farms delivering flocks positive for Campylobacter spp., and surveys of broiler meat products at the retail level. This article presents results covering the inclusive 3-year period between 2002 and 2004. During this period, a total of 10,803 flocks from 562 broiler farms were tested; altogether, 521 (4.8%) of the flocks were identified as positive for Campylobacter spp., primarily Campylobacter jejuni. The positive flocks originated from 257 (45.7%) of the farms. During the period 2002 to 2004, there was a large and steady reduction in flock prevalence, from 6.3% in 2002 to 3.3% in 2004. Also, the proportion of farms producing flocks positive for Campylobacter spp. each year reduced substantially, from 28.4% in 2002 to 17.8% in 2004. The proportion of flocks positive for Campylobacter spp. varied considerably with season and region. The action plan is a successful collaboration between academia, regulatory agencies, and the poultry industry that has resulted in a significant reduction in the number of broiler carcasses positive for Campylobacter spp. on the market. The temporal associations between implementation of the control program and the drop in the number of infected chickens and contaminated carcasses indicate that this collaborative action plan has been instrumental in achieving the goals of enhancing food safety. PMID- 16245735 TI - Prevalence of Shiga toxin-producing Escherichia coli in beef cattle. AB - A large number of Shiga toxin-producing Escherichia coli (STEC) strains have caused major outbreaks and sporadic cases of human illnesses, including mild diarrhea, bloody diarrhea, hemorrhagic colitis, and the life-threatening hemolytic uremic syndrome. These illnesses have been traced to both O157 and non O157 STEC. In a large number of STEC-associated outbreaks, the infections were attributed to consumption of ground beef or other beef products contaminated with cattle feces. Thus, beef cattle are considered reservoirs of STEC and can pose significant health risks to humans. The global nature of the human food supply suggests that safety concerns with beef will continue and the challenges facing the beef industry will increase at the production and processing levels. To be prepared to address these concerns and challenges, it is critical to assess the role of beef cattle in human STEC infections. In this review, published reports on STEC in beef cattle were evaluated to achieve the following specific objectives: (i) assess the prevalence of STEC in beef cattle, and (ii) determine the potential health risks of STEC strains from beef cattle. The latter objective is critically important because many beef STEC isolates are highly virulent. Global testing of beef cattle feces revealed wide ranges of prevalence rates for O157 STEC (i.e., 0.2 to 27.8%) and non-O157 STEC (i.e., 2.1 to 70.1%). Of the 261 STEC serotypes found in beef cattle, 44 cause hemolytic uremic syndrome and 37 cause other illnesses. PMID- 16245736 TI - [Intravitreal administration of drugs]. AB - The direct intravitreal administration represents the easiest and quickest method for obtaining efficient substance concentrations closer to the lesions and is reserved to the pathology which is resistant to the common treatments. PMID- 16245737 TI - [Duane's syndrome--etiopathogenesis, clinical features and diagnosis]. AB - Duane syndrome is a congenital disease also known as congenital retraction syndrome or Stilling-Turk-Duane retraction syndrome. There are three types of Duane syndrome; their variability is given by the aberrant innervation of the lateral rectus muscle. The symptoms of the disease were described and they are all known, but the etiology and the pathology remain unclear. Although Duane syndrome was considered as a pure local myogenic phenomenon (the lateral rectus muscle fibrosis), the modern theory is the absence of the abducens nuclei. Various theories have been put forward on the basis of data collected from surgical, autopsy and electromyography studies. The most frequent and terrifying differential diagnosis is the VI nerve palsy and the symptoms of Duane syndrome suggest an intracranial disease, so we understand the parents and patient fear. That's why is very important to explain them the causes and the consequences of the disease and to educate the patients to obtain the optimal functional and mental comfort. Finally, we have in mind one, at least subjective, question: why is the abducens nerve nucleus affected and how? PMID- 16245738 TI - [Immunoprophylaxis versus immunotherapy in herpetic eye infections]. AB - The preventive and curative treatment of the eye herpetic infections involves, besides the antiviral therapy, also different approaches which can stimulate the natural protective capacity of the human body (the antiherpetic vaccine and the immunomodulation agents). PMID- 16245739 TI - [The role of tumor necrosis factor in inflammation]. AB - The paper presents up-to-date information about the action of the tumor necrosis factor in inflammatory pathology. It underlines the fact that a new category of medicines appeared: anti-TNF monoclonal antibodies which can offer new therapeutic perspective in the treatment of intraocular inflammation. PMID- 16245740 TI - [Principles of treatment in ocular burns regarding the ocular surface and limbal stem cells]. AB - The term ocular surface emphasizes the functional interdependence of the nonkeratinizing epithelium of cornea and conjunctiva. The limbal stem cells are responsible for replacement of corneal epithelium following ocular surface injuries. Over the past decades important advances in the management of chemical injury have occurred based on the application of theories on ocular surface and limbal stem cells. PMID- 16245741 TI - [Recurrent chronic uveitis in oligoarticular juvenile chronic arthritis--case report]. PMID- 16245742 TI - [Exophthalmos--case report]. AB - Exophthalmos is one of the most common clinical manifestations of an orbital abnormality. It usually results from a mass, a vascular abnormality, or an inflammatory process. We discuss the difficulty to obtain a correct diagnosis and the applied treatment in a clinical case, and at the same time we remark the importance of the close collaboration between the ophthalmologist and other specialists. PMID- 16245743 TI - [Keratotic basal cell carcinoma with orbital extension--case report]. AB - This is a description of a case of a 72-year-old female who developed a basal cell carcinoma of the external angle of the eyelid, with intraorbital extension. This type of cancer of the eyelid developed on keratotic skin lesions of the face; the invasion of the orbit occurred only for two years. The tumour of the eyelid and orbit was completely removed with good postoperative course and a good outcome without recurrence in the past 3 years. PMID- 16245744 TI - [Pathological cataracts in systemic diseases]. AB - PURPOSE: To show the physiopathological mechanisms which determines the appearance of the pathological cataract and types of cataract and clinical manifestations of the systemic disease and operative complications. METHOD: Retrospective study that includes 22 patients hospitalized in clinic with pathological cataract diagnosis, appeared in following systemic disease: diabetes mellitus, skin disease, endocrine disease, SNC disease. RESULTS: Generally the patients had posterior subcapsular cataract or nuclear and posterior subcapsular cataract with fast evolution to intumescent cataract. Phacoemulsification with IOLCP performed at all patients. Good evolution and not surgery complications. CONCLUSIONS: The difficult of the surgery was connected with the risk of the posterior capsular break ( the majority cases were soft cataract) and also connected with the capsulorhexis making in intumescent cataracts. The post surgery evolution was not different compared with a usually cataract. PMID- 16245745 TI - [Congenital retinal folds in different clinical cases]. AB - We present 12 clinical cases of congenital retinal folds with different etiologies: posterior primitive vitreous persistency and hyperplasia (7 cases),retinocytoma (1 case). retinopathy of prematurity (1 case), astrocytoma of the retina (1 case), retinal vasculitis (1 case), Goldmann-Favre syndrome (1 case). Etiopathogenic and nosological aspects are discussed; the congenital retinal folds are interpreted as a symptom in a context of a congenital or acquired vitreo-retinal pathology. PMID- 16245746 TI - [Research about surgical treatment in primary congenital glaucoma]. AB - The aim of this investigation was the research of different types of surgical treatments applied to the early primary congenital glaucoma, and the identification of the best and most efficient therapies for each individual. It is a retrospective research on a number of 74 eyes with early primary congenital glaucoma, who were diagnosed and had an operation during the first four years from birth, since 1990 to March 2004, in 6 hospital centres from Romania. There were 101 surgical operation, the most frequent surgical methods used were trabeculotomy and trabeculectomy, which produce similar effects. PMID- 16245747 TI - [Pressure stress in glaucoma--assessment through classical techniques and computational modelling]. AB - Even if glaucoma remains a major eye illness with unknown etiology, the importance of vascular factors associated with mechanical factors contributing to the pathogenesis and development of glaucomatous optic neuropathy has been emphasized. the objective of the study consists in elaboration of a biomechanical model of the human eye correlated with clinical and ultrasonography evaluations aiming to early identification of changes induced by pressure stress in open angle glaucoma (OAG) at retrobulbar arteries and optic nerve head level. METHOD AND MATERIAL: The present study included a lot formed by 97 patients without glaucoma, aged 20-75 years (lot 1) and a lot formed by 71 patients with open angle glaucoma, aged 45-80 years (lot 2). We used usual methods in ophthalmological examinations and diagnosis (bottom ocular fundus examinations eye examinations, visual - field examinations, measurements of PIO), ultrasonography methods (CDI, pulsed Doppler, spectral Doppler and power-angio), computational modelling methods for ocular structures. RESULTS AND CONCLUSIONS: In open angle glaucoma color Doppler imaging shows changes in retrobulbar hemodynamic for most of the studied cases, ultrasonography techniques being considered as an important tool for the diagnosis and progression control in glaucomatous damage. Limitations of ultrasonography techniques consist of early identification of these alterations and can be improved by the elaboration of a biomechanical model of the human eye that will permit an abstraction and generalisation of the hemodynamic aspects with the aim of simulation of changes induced by high intraocular pressure on retrobulbar arteries level, the possibility of viewing these changes in their dynamics (on lower pressure values than 30 mmHg). PMID- 16245748 TI - [Diagnosis of rheumatoid uveitis in children and teenagers]. AB - We have done a retrospective study of juvenile rheumatoid uveitis of hospitalized cases between 1993-2002. 96 juvenile uveitis were available and 18 of this cases were diagnosed with juvenile rheumatoid uveitis. Juvenile rheumatoid arthritis associated uveitis is one of the most difficult of the uveitic entities to manage. Juvenile rheumatoid arthritis is the most common rheumatic disease of childhood. The classic presentation is an asymptomatic, bilateral, non granulomatous iridocyclitis and the latency between onset of arthritis and detection of uveitis is around two years. Arthritis precedes the onset of uveitis, so the ophthalmologist will usually see the patients after the diagnosis has been established by the pediatrician or rheumatologist. PMID- 16245749 TI - [Advantages and disadvantages of contact lenses in the correction of ametropia]. AB - PURPOSE: The purpose is to appreciate the benefits and the drawbacks in the correction of the refractive errors with contact lenses comparative to the glasses, to see their indications, general contraindications, their specific features. MATERIAL AND METHOD: This is a retrospective study on a group of 100 patients that were consulted during a period between years 2003 and 2004 in the Optimed Clinic of Ophthalmology. RESULTS: The patient's age varied between seven to sixty years, most of them between twenty and thirty years. Women formed the largest group most of the patients lived in the cities. The refractive errors in which we used contact lenses were: myopia, astigmatism, hyperopia, keratoconus. Soft lenses were used more then hard lenses. Only 25 of our patients developed some complications, especially corneal complications. CONCLUSIONS: 1. The contact lenses represent an optimal solution in the correction of refractive errors, especially in young patients. 2. The quality of vision obtained with contact lenses is superior comparative to glasses. 3. Most of their complications can be treated without marks. PMID- 16245750 TI - [Treatment of choroidal neovascular membranes]. AB - PURPOSE: The purpose is to present our experience in diagnosis and treatment of choroidal neovascular membranes of various etiology. MATERIAL AND METHODS: This is a retrospective study consisting of 165 patients that were diagnosed with choroidal neovascular membranes (CNM). From this patients, 62 were treated between 2001-2005 for CMN of different etiology: age-related macular degeneration (AMD), myopia, presumed ocular histoplasmosis syndrome, angioid streaks, viteliform dystrophy. The diagnosis was based on the clinical examination (direct and indirect ophthalmoscopy, direct and indirect biomicroscopy) and angiofluorography. The treatment was either PDT for sub-foveolar membranes, or laser coagulation with YAG532 laser for juxta and para-foveolar well defined membranes, or combined treatment for large minimal classic membranes with a well defined juxta para-foveolar component or a component on the feeder vessels. PDT was done with verteporfin (Visudyne, Novartis Pharma AG) activated with a diode laser of 689 nm (Activis, Quantel medical) and the laser coagulation was done with a Visulas YAG 532 Carl Zeiss laser. RESULTS: Results can be evaluated from two points of view: changes in the neovascular membrane (enlargement and activity signs) and the changes in VA. Last researches are based not only on AV but also on the quality life. The membranes are closed with PDT in 90% of the cases in the first week but there are rebounds in almost 100% of the cases at 3 months. This calls for multiple treatments. After two or three treatments the signs of activity (hemorrhage and exudation) drop and 60% of the membranes will close. After laser coagulation the CMN is closed completely in 70% of the patients and in 20% of the cases there is a rebound in the first month and in 30% of the cases the rebound appears in the next 2 years. The visual acuity has improved at 33% of cases, remained the same in 33% of cases and worsened in the rest of the treated eyes. The results are compatible with TAP and VIP results where after 24 months the average VA of those treated with verteporfirin was lower than at the beginning but twice as good compared with placebo. CONCLUSIONS: For four year we had at our disposal methods with which we can improve (33%) the VA in patients with CMN, at some of them we could maintain (33%) the visual acuity and at the rest of the patients, even though the visual acuity is lower, it will be still higher then the one during the natural course of the disease. PMID- 16245751 TI - [Morphopathological study in age related macular degeneration]. AB - The aim of this paper is to study the morphopathological changes manifested in age-related macular degeneration. MATERIAL AND METHOD: The researched histopathological material consisted in two eye balls enucleated because of irreversible eye diseases from aged persons suffering from age-related macular degeneration. The method applied for processing the material was inclusion in paraffin, followed by usual coloration with hematoxylin-eosin. RESULTS AND CONCLUSIONS: The established changes (hard drusen, soft drusen, calcified drusen, changes of Bruch's membrane, subretinal neovascularization, serous retinal pigment epithelial detachment, disciform degeneration) prove the importance of Bruch's membrane in the pathogenesis of age-related macular degeneration. PMID- 16245752 TI - Towards the origin of the term physiotherapy: Dr Edward Playter's contribution of 1894. PMID- 16245753 TI - The effect of angle and oscillation on mucous simulant speed in flexible tubes. AB - BACKGROUND AND PURPOSE: The aim of this study was to investigate, in a tube model, how the speed of a mucous simulant was influenced by angle and different types of oscillations. METHOD: Using a repeated-measures study design, the primary outcome measure was the mucous simulant speed calculated from the time taken for the mucous simulant to travel a distance of 10 cm. Ultrasonic gel diluted to a viscosity (113 Poise), approximating human sputum, was introduced into a flexible tube similar in diameter to the human adult trachea. The tube was subjected to discrete angles of 0 degrees and 30 degrees, 60 degrees and 90 degrees downward. Symmetrical oscillation was applied in both the transverse and longitudinal directions with frequencies of 5, 15 and 25 Hz at amplitudes of 1 mm and 2 mm peak-to-peak using a commercially available oscillator. Asymmetrical oscillation was applied using repeated cycles of slow acceleration and fast deceleration in the longitudinal direction at 0 degrees and 30 degrees downward and up a 5 degrees incline using a custom-built apparatus. RESULTS: Each 30 degrees angle increment of the tube from 0 degrees to 90 degrees significantly increased mucous simulant speed (p<0.001). Symmetrical oscillation did not provide an advantage over angle in terms of mucous simulant speed; however, asymmetrical oscillation increased mucous simulant speed beyond that caused by angle for all angles tested (p<0.001) and was able to drive mucous simulant up a small incline (5 degrees) in this tube model. It was found that certain types of longitudinal oscillation elongated the mucous simulant. CONCLUSIONS: The present study supports the use of gravity to assist in secretion clearance. Asymmetrical oscillation is a novel technique which warrants further investigation. PMID- 16245754 TI - Use of devices to prevent subluxation of the shoulder after stroke. AB - BACKGROUND AND PURPOSE: Supportive devices, such as slings, wheelchair or chair attachments and orthoses, have been used for many years by physiotherapists to support the affected shoulder after stroke. The purpose of the present paper was to examine the alignment between current practice with evidence for the use of supportive devices in the prevention of subluxation of the shoulder after stroke and to provide guidance for clinical practice and future research. METHOD: In order to determine the evidence regarding the most appropriate supportive devices to prevent subluxation after stroke, a systematic review was undertaken following the guidelines set out by the Cochrane Collaboration. In order to determine current practice with regard to the most commonly used supportive devices to prevent subluxation after stroke, a questionnaire was designed and administered for use in Australia. RESULTS: The collar-and-cuff sling is the most commonly used sling. However, a small amount of lower-level evidence shows that the collar and-cuff sling only reduces subluxation by half, suggesting that it may not be the most effective to use for prevention. In contrast, this evidence supports the use of wheelchair or chair attachments. CONCLUSIONS: We found that there was a lack of high-level evidence to guide clinical practice. In order to determine evidence-based practice for the prevention of shoulder subluxation, there is a need to test the efficacy of the most promising supportive devices based on available evidence. PMID- 16245755 TI - Postural instability during reaching tasks in Parkinson's disease. AB - BACKGROUND AND PURPOSE: Reaching, like turning and rising from sitting, is an activity commonly associated with falls by people with Parkinson's disease. We set out to: (a) identify how people with moderate and severe Parkinson's disease performed high and low reaches and (b) to explore in detail the standard functional reach (functional reach) test and the fall histories of those who appeared unstable when reaching. METHOD: In this cross-sectional study, people with moderate or severe Parkinson's disease (Hoehn and Yahr Grade III or IV) were video-recorded at home, reaching above shoulder height and below knee level. Blinded observers rated stability, alignment, foot position, floor contact, use of support and reach type. Functional reach was also measured and participants were interviewed about falls. RESULTS: Of 51 participants, 33 (65%) had moderate Parkinson's disease and 18 (35%) severe. A greater proportion of the latter used support when reaching high (p = 0.029) and aligned forward when reaching low (p = 0.015); otherwise, strategies were similar across groups. Six people (all with severe Parkinson's disease) appeared unstable when reaching: they had a shorter functional reach than the others (median 10 cm versus 18 cm; p = 0.042) and had fallen frequently (median five falls in a year), although rarely when reaching. CONCLUSIONS: Reaching tasks challenge postural stability in severe Parkinson's disease. People who appear unstable when reaching are likely to be repeat-fallers and at risk of further falls during more demanding activities. Research should address whether discouraging potentially destabilizing manoeuvres (such as squatting and toe-standing) and promoting safety-enhancing strategies (such as using support and facing forward), with or without balance retraining, reduces the risk of falling among people with severe Parkinson's disease. PMID- 16245756 TI - Re-admission to intensive care: identification of risk factors. AB - BACKGROUND AND PURPOSE: The re-admission of patients to intensive care is associated with increased morbidity, mortality, loss of morale for patients and family, and increased health costs. The aim of the present study was to identify factors which place patients at a higher risk of re-admission to intensive care. METHOD: A prospective study of patients who were re-admitted to a 22-bed tertiary level intensive care facility within a 12 month period. Data were kept on every patient re-admitted to intensive care, including standard demographic data, initial admission diagnosis, co-morbidities, re-admission diagnosis, mobility on discharge, secretions, airway, chest X-ray, PaCO2, PaO2, PaO2/FiO2 and time of discharge. Subjects included 74 patients who had been re-admitted to intensive care in 12-month period and a comparison group of patients who were not re admitted to intensive care. A cross-tabs procedure was initially used to estimate maximum likelihood. Significant factors with an alpha value of <0.10 were entered into a backward conditional binary logistic regression to determine independent factors. RESULTS: The overall percentage of patients who were re-admitted to intensive care compared with total admissions was 7.7%. Significant independent factors for re-admission were found to be age >65 years (p<0.05), colonization (p<0.001), weakness (p<0.001), co-morbidities of cardiac and/or respiratory disease (p<0.001) and depression (p<0.001). CONCLUSIONS: A certain profile of patients has been established who are at increased risk of re-admission to intensive care. These patients could be provided with increased intervention and surveillance on discharge from intensive care. PMID- 16245757 TI - Case report: physiotherapy strategies for a patient with conversion disorder presenting abnormal gait. PMID- 16245758 TI - Case report: the effect of physiotherapy and orthotic intervention, 40 years after stroke. PMID- 16245759 TI - The pusher syndrome in a patient with cerebellar infarction. PMID- 16245760 TI - [Prevalence of endodontic yeasts in periapical infections]. AB - In literature, microbiology results of periapical lesions are limited. Only several types are frequently isolated in rebellious cases. These ones include Enterococcus Faecalis and Enterococcus gram. Recently, a gain of interest is focused on discussion concerning endodontic yeasts that is why we developed our research in this way. From the 81 cases of endodontic culture studied, 20% were positive isolating Candida in which 14% weren't C. albicans, for the rest we identify C. glabrata, C.sp., C. kefyr, C. kruseii. The predominance of Candida albicans was justified by their frequency as commensal oral flora, explaining someway the failure of some intracanal endodontic medication. PMID- 16245761 TI - [The treatment of choice for recurrent ameloblastoma]. AB - 92 patients with recurring ameloblastoma of jawbones were followed. The result after submitting these patients under preserving surgical therapy were evaluated on the basis of compared biostatistical and bioclinical criterion. Among these 92 patients, 66 were Negro African series with a success rate of 82% and 26 others were white "Caucasian" series with a success rate of 85%. What implies that the preserving surgery on the recurring ameloblastoma is more beneficial than negative with the two races with more than 80%. The authors thus concluded that this surgical method still keeps its place of choice in the treatment of the repetitions of the ameloblastoma and that this factor has a prognostic value. PMID- 16245762 TI - [Cephalometric study of cranio-facial growth of 902 Black African children and adolescents aged 8 to 20]. AB - Cephalometric studies of the growth are numerous and represent a contribution to the field of the anthropological knowledge of the stages of the cranio-facial growth and for a better adaptation of the orthodontic treatments. However, in Black Africa people there are very few studies of this kind. The purpose of this investigation is to analyze the growth of the cranial base, the maxillary skeletal region and mandibular skeletal region at the young African subjects. The sample of study consists of 386 girls and 516 boys in period of growth, old from 8 to 20 years, presumedly orthomorphic and having lateral cephalometric radiograph in occlusion. The statistical analysis of the data recorded on the layouts cephalometric (test U of Mann Whitney) announces the existence of a sexual dimorphism which is strongly significant around 9 years, 11 years and 17 years. Maxilla and mandible present different rates of growth while remaining active up to 20 years but the growth of the base of cranium seems to be stabilized earlier, after 11 years. The taking into account of these morphological variations are determining for the conduit of treatments more stable and adapted to our populations. PMID- 16245763 TI - [Prevalence of rhino-pharyngeal disease in the presence of malocclusion in school children in the village of Abidjan]. AB - An epidemiological survey of randomly selected school children led to Abidjan in three public schools made it possible to determine the prevalence of the rhinopharyngeal diseases in presence of malocclusions. It is an exploratory study jointly undertaken by two teams of specialists in ORL and Orthodontics within a sample of African schoolchildren old of 5 to 21 years. The results showed the presence of malocclusions in 73.30% of the cases (N = 220). The rhinopharyngeal diseases account for 38.3% (N = 115). At the subjects carrying a malocclusion the rhinopharyngeal diseases are 48.30% and they are dominated by the allergic chronic rhinitis (40%) followed obstructive hypertrophic tonsillitis (16.5%). However, these states which cause certainly a nasal obstruction involving an oral breathing do not cause inevitably malocclusion. We cannot thus affirm unambiguous a bond between malocclusions and rhinopharyngeal diseases (p > 0.05). They are nevertheless as many indications to question the patient on other symptoms and to refer to an ORL specialist. Our investigation being limited to the occlusal study, it seems essential to us to continue these analyses to detect the possible predisposition of certain patients to develop dento-skeletal anomalies in the presence of rhinopharyngic diseases. PMID- 16245764 TI - Expectations of orthodontic treatment and satisfaction with dental appearance among Nigerian parents [corrected]. AB - The aim of this study was to investigate correlations between satisfaction with dental appearance and expectation of orthodontic treatment. The effects of sex and age on these variables were also explored. A sample of 466 parents comprising 238 (51.1%) males and 228 females (48.9%) were randomly surveyed in Ibadan Nigeria. Their ages ranged from 18 to 73 with mean age of 36.89 +/- 12.27 (SD). They were asked whether they were satisfied or otherwise with their dental appearance and to indicate their expectation of orthodontic treatment. A multiple regression analysis was used to estimate the effect of the dental satisfaction on expectation of orthodontic treatment as well as the influence of age and sex on orthodontic expectations. Significant correlation was not found between satisfaction between dental appearance and parents' expectations. These correlations were invariant over gender, but not over age. It was concluded that satisfaction with dental appearance is not a significant predictor of parents' expectations of orthodontic treatment among the study sample but age was. PMID- 16245765 TI - [Comparative study of 2 antibiotic combinations in the treatment of rapidly progressing periodontitis]. AB - To get rid efficiently of anaerobics bacteria, which constituted the flora associated to periodontitis, the periodontal therapy is now moving towards 2 or 3 antibiotics association. The aim of this study is to make a comparison between the efficacy of the association of amoxicillin plus clavulanic acid and that of amoxicillin plus metronidazole into 20 patients suffering from rapidly progressive periodontitis. The results have shown that both forms of antibiotics association are efficacious in rapidly progressive periodontitis treatment, with the same action on clinical parameters. PMID- 16245766 TI - [Cardiac patient. When may he have sex?]. PMID- 16245767 TI - [Safe start, adequate training. How to mobilize your heart patients]. PMID- 16245768 TI - [How much exercise must the heart patient have? "2,000 kilocalories per week is the minimum"]. PMID- 16245769 TI - [Beginning therapy of heart failure with beta blockers. Do these patients survive longer?]. PMID- 16245770 TI - [Finely adjusted differential therapy succeeds this way! References for dose adjustments in heart failure (interview by Dr. Dirk Einecke)]. PMID- 16245771 TI - [ASCOT-BPLA Study raises new questions regarding: which is the best initial antihypertensive drug?]. PMID- 16245772 TI - [Every 10th person has actinic keratosis. Cold, light and cream prevent skin cancer]. PMID- 16245773 TI - [Etiology of persistent joint instability. Unstable knee despite cruciate ligament surgery--a medical error? (interview by Dr. Thomas Meissner)]. PMID- 16245774 TI - [Visits to the home for the aged and new EBM. Now we must correct the numbers for weeks!]. PMID- 16245775 TI - [Social law contradicts economic regulation. Home care patient dead--tablets in the trash]. PMID- 16245776 TI - [Acute HIV infection is seldom recognized]. PMID- 16245777 TI - [Chronic inflammation of the upper airways. Operation instead of antibiotic]. PMID- 16245778 TI - [New information on chronic rhinosinusitis and polyposis nasi]. AB - New Information on Chronic Rhinosinusitis and Polyposis Nasi Chronic inflammation of the paranasal sinuses is more common than inflammatory or degenerative diseases of the joints or arterial hypertension. The pathogenesis of chronic rhinosinusitis has still not been completely worked out. It is known that in particular the administration of antibiotics is incapable of healing the condition. Once the diagnosis has been established by an ENT specialist, topical or systemic steroids may be helpful. In most cases, however, it is necessary to combine these substances with surgical treatment. For this purpose, endoscopic sinus surgery is an approach that is capable of providing good long-term results. PMID- 16245779 TI - [Chronic tonsillitis--pathogenesis, symptomatology and therapy]. AB - Chronic tonsillitis is a common disease entity which, on account of the possibility of the tonsils becoming a focus of infection, must not be made light of. The patient's complaints are highly uncharacteristic, and it is not always possible to establish the diagnosis on the local findings alone. Rather, the patient's history and general state of health must also be considered when considering the diagnosis. By reason of the pathogenesis, the treatment of choice can only be tonsillectomy. PMID- 16245780 TI - [General use of vitamin supplementation]. AB - Current scientific wisdom has it that the general supplementation with vitamins is unnecessary. For certain risk groups, such as infants, pregnant women and the aged,the prophylactic administration of vitamins may be useful. Otherwise,vitamin supplementation of the diet may have a positive impact on health at most in persons with a suboptimal vitamin supply via their usual nutrients. A supplementation should be preceded by a history-taking (e.g. via a dietary protocol). Over the long-term, pharmacological doses of vitamins can give rise to adverse side effects. PMID- 16245781 TI - [Local administration of essential oils in the common cold. No risk for the skin]. PMID- 16245782 TI - [Fine dust alarm. More deaths due to air pollutants than traffic accidents]. PMID- 16245783 TI - [Still tougher bandages for control of drug costs. Defending yourself against regress fees]. PMID- 16245784 TI - [Hypertensive patient consults for lumbago. Treatment complex for chronic patients is still reimbursable]. PMID- 16245785 TI - [First antibody against IgE in view. Hope for severely ill asthmatic patients ]. PMID- 16245786 TI - [Deviated tongue]. PMID- 16245787 TI - [Dysphagia--differential diagnosis and therapy. What is behind a lump in the throat?]. AB - Swallowing disorders are the symptom of numerous conditions. The diagnostics and therapy require the close interdisciplinary collaboration of different medical specialists. PMID- 16245788 TI - [Totally healthy despite muscle atrophy?]. PMID- 16245789 TI - [Improvement on chest X-ray findings after two months of antituberculous treatment]. AB - PURPOSE: To detect factors that affected the improvement on chest X-ray findings after two months of antituberculous treatment. METHODS: We assessed the chest X ray findings of 72 patients with sputum culture positive to determine whether findings showed improvement after two months treatment with antituberculous drugs, and we compared the differences in disease status between the improved group (I group) and the non-improved group (N group). RESULTS: The I group consisted of 43 pts. and the N group consisted of 29 pts. The mean age in the N group was higher than that in the I group. There were no differences in sex or body weight. Though the mortality rate in the N group was 20.7% and that in the I group was 9.3%, it was not statistically significant. The erythema size of tuberculin skin test in the I group was larger than that in the N group. There were no differences in the type or extent of lesion on Gakkai classification between the two groups, and there were no remarkable blood biochemistry findings in either group. Though there were no differences in the rates of side effects or discontinuation of treatment, the incidence rate of complications in the N group was higher than that ih the I group. CONCLUSION: Older patients with complications and smaller erythema size of tuberculin skin test reaction showed a delay in chest X-ray findings improvement after two months of antituberculous drug treatment. PMID- 16245790 TI - [The improvement of the enforcement rate of the standard regimen (A) of tuberculosis chemotherapy by the introduction of common database system for tuberculosis]. AB - OBJECTIVE: The purpose of this study was to improve the enforcement rate of the standard regimen (A) of tuberculosis chemotherapy. SUBJECTIVE AND METHODS: We introduced the common database system for tuberculosis in three national hospitals in Hokkaido. From January 2002 to December 2003, we collected the anonymous informations of the patients with tuberculosis at the start of treatment, at the discharge and at the end of treatment. Then, we reported the enforcement rate of the standard regimen (A) as a clinical indicator periodically to three hospitals. RESULTS: Four hundred and twenty-nine patients were registered. In patients below 80 years old, the enforcement rate of the standard regimen (A) was 48.5% in 2002. The enforcement rate rose significantly to 62.7% (p = 0.0126) in 2003. In elder smear-positive patients (> or =75) and in elder smear-negative patients (> or =70), the enforcement rate was low (29.1% and 25.0%, respectively). Furthermore in young smear-negative patients (< or =29), the enforcement rate was low (28.0%). As the extent of their disease was minimal, they were treated with other regimens. In patients treated with the standard regimen (A), there were no significant differences in the frequency of adverse effects between elder patients ( 70) and other patients (< or =69). There were also no significant differences in the frequency of changing the regimen between them. Median admission period of 2002 was 114 days. In 2003, it was shortened significantly to 110 days (p = 0.0487). CONCLUSION: By the introduction of the common database system for tuberculosis, the enforcement rate of the standard regimen (A) was improved. Low enforcement rate in young smear-negative patients in an important problem to be improved in the future. The clinical indicator based on the common database system between hospitals, is useful to clarify the problems, and then to improve the quality of medical performance. PMID- 16245791 TI - [A case of Mycobacterium intracellulare lung disease that was detected by health examination and was successfully cured by chemotherapy--including discussions on the diagnostic criteria of the disease]. AB - A case was 56 years old woman, and she did not have any subjective symptom. She received multiphasic health screening, and abnormal shadow was detected on her chest radiograph. Chest radiography revealed infiltrations in the middle lobe. Computed tomography (CT) of the thorax showed clusters of small nodules in the middle lobe. The bronchial washing specimen showed acid-fast bacilli identified as Mycobacterium intracellulare by DNA-DNA hybridization (DDH) method. This case was diagnosed as Mycobacterium intracellulare lung disease. The patient received combination therapy with rifampicin, ethambutol, and clarithromycin for one year with radiological improvement. CT findings were characteristic and useful for the early diagnosis of MAC infection, which led to cure of the disease by chemotherapy. PMID- 16245792 TI - [Anti-tuberculosis drug-induced hepatitis]. AB - In 1996 a six-month short-course chemotherapy was adopted as a standard chemotherapy for pulmonary tuberculosis in Japan. The frequency of implementation of short course chemotherapy for smear positive new case was only sixty percent in 2003, although a short course chemotherapy was recommended all over the world for avoidance of acquiring a new resistance among new drug resistant tuberculosis. The reasons for the low adaptation of short course chemotherapy were speculated that high rate of elderly patients, and high frequency of drug induced hepatitis. From our 14-year experience, the frequency of drug-induced hepatitis was 7.8% for all patients with standard 6-month or 9-month regimen, 7.2% in the patients with normal liver function tests at the start of chemotherapy, and 11.8% in the patients with any kinds of abnormality. The death rate was 0.04% among treated patients, and fatality was 0.49% among the patients with drug-induced hepatitis. Positive HCV antibody and less than 1,000 cells/microL of peripheral lymphocyte count at the beginning of treatment were independent risk factors for drug-induced hepatitis. The management of hepatitis during antituberculosis treatment was also referred. PMID- 16245793 TI - [Frontier of mycobacterium research--host vs. mycobacterium]. AB - During the past decade, we have observed advance in tuberculosis research including novel vaccines, innate immunity (TLR), SNIP analysis and molecular mechanism of drug resistance. Worldwide genome project enabled the whole genome sequence of host resistant against tuberculosis as well as the whole genome sequence of M. tuberculosis H37Rv. DNA technology has also provided a great impact on the development of novel vaccine against TB. In this symposium, we have invited leading researchers in the field of the frontier study of Mycobacterium research in order to provide general overview of the cutting edge of frontier research. Molecular mechanism of drug resistance of M. tuberculosis has been clarified. On the other hand, molecular mechanism of host-defence (insusceptibility of host) against M. tuberculosis has not yet elucidated. Dr. Taro Shirakawa (Kyoto University) reviewed the susceptibility genes of host in TB infection and presented candidate genes associated with multi-drug resistant tuberculosis. Dr. Naoto Keicho (International Medical Center of Japan) tried to identify host genetic factors involved in susceptibility to pulmonary Mycobacterium avium complex (MAC) infection by candidate gene approach and genome wide approach. In Japan, Dr. Masaji Okada (National Hospital Organization Kinki Chuo Chest Medical Center) has been engaged actively in the development of new tuberculosis vaccines (HVJ-liposome/Hsp65 DNA + IL-12 DNA vaccine and recombinant 72f BCG vaccine). He showed basic strategy for construction of new candidate vaccines and also showed significant efficacy on the protection of tuberculosis infection using cynomolgus monkeys, which are very similar to human tuberculosis. Dr. Hatsumi Taniguchi (University of Occupational and Environmental Health) presented that M. tuberculosis mIHF and the neighbor genes went into a dormacy like state of M. smegmatis in J774 macrophage cells. This study might provide a weapon for elucidating the mechanism of dormacy of M. tuberculosis and the development of novel therapy. Dr. Chiyoji Abe (Nippon Becton Dickinson Co.) reviewed the molecular basis of the resistance to anti-tuberculosis drugs. Most cases of resistance are related to simple nucleotide substitutions rather than to acquisition of new elements. Dr. Kiyoshi Takeda (Kyushu University) showed interesting finding. He analyzed whether Toll-like receptor (TLR)-mediated activation of innate immunity in host defense against mycobacterial infection. MyD88/TRIF double defi-indicating that innate immunity is involved in anti mycobacterial infection. (1) SNP (single nucleotide polymorphism) analysis in association with Mycobacterium tuberculosis: Taro SHIRAKAWA (Department of Health Promotion & Human Behavior, Kyoto University Medical School, and RIKEN SRC Center) Candidate gene approach was made on 18 SNPs in 11 genes in association with M. tuberculosis. Patients with multi-drug resistance against M. tuberculosis are also subjected. SNPs in NRAMP1 gene were associated with the disease, and drug resistance, its mechanisms remain unknown. (2) Search for genes susceptible to pulmonary Mycobacterium avium complex infection: Naoto KEICHO (Department of Respiratory Diseases, Research Institute, International Medical Center of Japan) Interaction among pathogens and host factors is important for development of infectious diseases. We are trying to identify host genetic factors involved in susceptibility to nonimmunocompromized pulmonary Mycobacterium avium complex (MAC) infection by candidate gene approach and genome-wide approach. Elucidation of functional significance of susceptibility gene polymorphisms will lead to a new strategy for control and prevention of the disease. (3) T cell immunity against Tuberculosis in host and the establishment of novel vaccine: Masaji OKADA (Clinical Research Center, National Hospital Organization Kinki-Chuo Chest Medical Center) T cell (CTL, Th1) immunity including granulysin play an important role in host defense against tuberculosis (TB) in human. Patients with TB or Multi-drug resistant TB showed suppression of all these immunities. HVJ liposome/Hsp65 DNA + IL-12 DNA vaccination was 100 fold more efficient than BCG on the elimination of Mycobacterium tuberculosis (M.TB) in the BALB/c mice. Cytotoxic T cells activity against M. TB was augmented. By using these new vaccines (Hsp 65 DNA + IL-12 DNA, recombinant 72f BCG) and the cynomolgus monkey models which are very similar to human tuberculosis, the prophylactic effect of vaccines was observed. Thus, these novel vaccines should provide a useful tool for the prevention of human TB infection. (4) Mycobacterium tuberculosis mIHF and the neighbor genes go into a dormancy-like state of M. smegmatis J15CS in J774 cells: Hatsumi TANIGUCHI (Department of Microbiology, School of Medicine, University of Occupational and Environmental Health) Mycobacterium smegmatis J15CS transformants harboring the mIHF gene or the mIHF-gmk-Rv1390 genes showed no difference in in vitro growth and acid-fastness. However, transformants harboring mIHF-gmk-Rv1390 formed short-rod cell morphology and decreased acid fastness in the mouse macrophage-like cell line J774 compared to those of the other transformants, and the nuclei of the infected J774 cells also changed. Nevertheless, the colony forming units were similar. These indicate that mIHF and the neighbor genes of M. tuberculosis might regulate a growth of mycobacteria in macrophages. (5) Molecular basis of the resistance to anti-tuberculosis drugs: Chiyoji ABE (Nippon Becton Dickinson Company, Ltd.) Considerable progress has been made toward understanding the molecular basis of the resistance to anti tuberculosis drugs. Most cases of resistance are related usually to simple nucleotide substitutions rather than to acquisition of new elements. Multi-drug resistant isolates of Mycobacterium tuberculosis arise a consequence of sequential accumulation of mutation conferring resistance to single therapeutic agents. The basis of resistance is not able to be explained yet in a substantial percentage of strains for other anti-tuberculosis drugs than rifampin and pyrazinamide. Further studies are required to fully understand the molecular mechanisms of resistance. (6) Toll-like receptors in anti-mycobacterial immune responses: Kiyoshi TAKEDA (Department of Molecular Genetics, Medical Institute of Bioregulation, Kyushu University) Toll-like receptors (TLRs) play an essential role in the recognition of specific patterns of microbial components. TLRs mediate activation of innate immunity and further development of antigen-specific adaptive immunity. In TLR signaling pathways, Toll/IL-1 receptor (TIR) domain containing adaptors, such as MyD88, TIRAP, TRIF, and TRAM, have been shown to play pivotal roles. Thus, the molecular mechanisms for TLR-mediated activation of innate immunity have been largely understood. We analyzed whether TLR-mediated activation of innate immunity is involved in host defense against mycobacterial infection. MyD88/TRIF double deficient mice, in which TLR-dependent activation of innate immunity is abolished, showed high sensitivity to mycobacterial infection, indicating that innate immunity is critically involved in anti-mycobacterial responses. PMID- 16245794 TI - Pharmaceutical data do not elude researchers. PMID- 16245795 TI - The environment isn't flat. PMID- 16245796 TI - Metals disturb seals' immune systems. PMID- 16245797 TI - Techniques help monitor microbial remediation. PMID- 16245798 TI - Dissolved organic matter may leave criminal fingerprint. PMID- 16245799 TI - No human risks with pharmaceuticals in water. PMID- 16245800 TI - Are membrane bioreactors ready for widespread application? PMID- 16245801 TI - Review of factors affecting microbial survival in groundwater. AB - This review quantitatively examines a number of published studies that evaluated survival and inactivation of public-health-related microorganisms in groundwater. Information from reviewed literature is used to express microbial inactivation in terms of log10 decline per day for comparison to other studies and organisms. The geometric mean value for inactivation rates for coliphage, poliovirus, echovirus, coliform bacteria, enterococci, and Salmonella spp. were similar at approximately 0.07-0.1 log10 day(-1), while geometric mean inactivation rates for hepatitis A virus, coxsackievirus, and phage PRD-1 were somewhat less at 0.02-0.04 log10 day( 1). Viruses show a temperature dependency with greater inactivation at greater temperatures; however this occurs largely at temperatures greater than 20 degrees C. Coliform bacteria die off in groundwater does not show the temperature dependency that viruses show, likely indicating a complex interplay of inactivation and reproduction subject to influences from native groundwater organisms, temperature, and water chemistry. The presence of native microorganisms seems to negatively impact E. coli survival more so than viruses, but in most cases, nonsterile conditions led to a greater inactivation for viruses also. The effect of attachment to solid surfaces appears to be virus-type dependent, with PRD-1 more rapidly inactivated as a result of attachment and hepatitis A and poliovirus survival prolonged when attached. PMID- 16245802 TI - Chemically activated luciferase gene expression (CALUX) cell bioassay analysis for the estimation of dioxin-like activity: critical parameters of the CALUX procedure that impact assay results. AB - The chemically activated luciferase gene expression (CALUX) in vitro cell bioassay is a bioanalytical tool that is increasingly being used by research and commercial laboratories for the screening and relative quantification of dioxins and dioxin-like compounds in sample extracts. Since CALUX analyses provide a biological response to all aryl hydrocarbon receptor active compounds present in a given sample extract containing a complex mixture of chemicals, interpretation of results is significantly more complexthan of chemical analyses. Operators in the laboratory can adjust many parameters when performing CALUX analyses, and the applied procedure strongly affects the result and, hence, the interpretation of the results. This paper examines critical methodological parameters and aspects of the CALUX bioassay that can affect the quality and accuracy of the analyses. Moreover, the study aims to identify the ways that alteration of these parameters influences CALUX measurements. A greater understanding of these characteristics will lead to increased accuracy, precision, and reproducibility of the widely used CALUX bioassay within and between research laboratories. PMID- 16245803 TI - Landfill-gas-to-Energy projects: analysis of net private and social benefits. AB - Methane emissions from municipal landfills represent 3% of the total United States greenhouse gas emissions that contribute to climate change. These methane emissions can be released to the air or collected and flared. This landfill gas also has the potential to be used to generate electricity. In 1994,the Environmental Protection Agency(EPA) created the Landfill Methane Outreach Program, with the objective of promoting the development of landfill-gas-to energy projects around the country. There are currently 2,300 active landfills in the United States. Although there are already 382 operational projects, there are many more landfills with the potential to use the gas. EPA has identified at least 630 candidate landfills for energy projects, and many more have still not been identified. The objective of this paper is to evaluate total private and social benefits of landfill-gas-to-energy projects, taking into consideration not only the costs of installing and maintaining the necessary equipment and the revenues obtained from selling the electricity but also a valuation of the greenhouse gas emissions that would be prevented and the emissions of criteria pollutants created by the electricity generating equipment. It also evaluates the breakeven government subsidies that would be required to make such projects economically viable from private and social perspectives in comparison to current subsidies. It was found that the private breakeven price of electricity for these projects is lower than dollar 0.04/kWh. Moreover, the optimum social subsidywas found to be less than dollar 0.0085/ kWh, which is about 40% lower than the currently available federal tax break of dollar 0.015/kWh. The method developed for this paper can be applied to other renewable energy technologies, to show their relative social costs and benefits. PMID- 16245804 TI - Influence of local human population on atmospheric polycyclic aromatic hydrocarbon concentrations. AB - Literature values of atmospheric polycyclic aromatic hydrocarbon (PAH) concentrations from sampling sites around the world were found, and using a high resolution human population grid, the population within a 25-km radius of each sampling site was calculated. A regression of concentration vs population revealed much about PAH concentration differences among regions as well as site locations within a continent. The best fit for the regression was for sampling locations in North America. A small amount of scatter was present for the regression of all developed countries indicating slight differences in emission regulations or energy usage. The regression from this plot was used as a benchmark for the expected relationship between PAHs and human population. Sites located within 25 km of a coasttended to have concentrations lower than expected, due to dilution with clean ocean air, while sites near industrial outputs or other point sources had higher than expected concentrations. Sites from developing countries typically had PAH concentrations that were far higher than those of the rest of the world. PMID- 16245805 TI - Congener-specific survey for polychlorinated biphenlys in sediments of industrialized bays in Korea: regional characteristics and pollution sources. AB - Areas of the Korean coastline with heavy industry and major harbors were investigated for polychlorinated biphenyl (PCB) pollution. This investigation paid attention to variations in the PCB congener patterns for a possible source of contamination. Surface sediments from 49 sites were sampled. Although the occurrence of PCBs in coastal marine environment correlates well with shipping and industrial activities, the contribution from shipping activities is considerable because of its enormous economical importance in Korea. The highest concentrations were found in harbors with heavy ship traffic and ship construction. Principal component analysis (PCA) of congener-specific composition of PCBs revealed distinct regional patterns, especially in a harbor and steel manufacturing area. PCB signatures with enhanced higher chlorinated congeners were typical for harbors with shipping activities and correlated well with commercial formulations that were formerly used in ship painting. Lower chlorinated congeners with up to five chlorines were significantly abundant in steel works zones which differed from harbor zones. This distinction was consistent with the congener patterns in the ambient air and the effluent of the steel works as well as in the nearby surface sediments. This study identified steel manufacturing as a recent and ongoing emission source of PCBs in Korea's coastal zone. PMID- 16245806 TI - Polychlorinated biphenyls in salmon and salmon feed: global differences and bioaccumulation. AB - Concentrations of 160 polychlorinated biphenyl (PCB) congeners or congener groups were determined in approximately 600 farmed Atlantic salmon from around the world and wild (ocean-caught) Pacific salmon from the Northeast Pacific. Concentrations and PCB congener profiles were analyzed to provide insight into the sources and uptake of PCBs in salmon as well as regional differences. Although total PCB concentrations in wild salmon appeared to be correlated to total lipid content, the increased proportion of total lipids in the farmed salmon could not account for the much greater PCB concentrations. We investigated the PCB congener patterns of hundreds of salmon samples using principal component analysis to further illuminate regional and species differences. Three major PCB patterns were observed, in most wild fish (except British Columbia and Oregon chinook), in farmed fish from the Atlantic, and in most farmed fish from the Pacific. The PCB congener profiles of farmed salmon often closely corresponded to a sample of commercial feed purchased in the same region, indicating that the feed is likely to be the major source of PCBs for farmed salmon. In such cases where PCB profiles in fish and feed were similar, a comparison of congener concentrations in fish and the feed showed that the majority of congeners, with some notable exceptions, were bioaccumulative to the same extent, irrespective of physical properties. PMID- 16245807 TI - An infrared and X-ray spectroscopic study of the reactions of 2-chlorophenol, 1,2 dichlorobenzene, and chlorobenzene with model cuO/silica fly ash surfaces. AB - The surface-mediated reactions of 2-chlorophenol, 1,2-dichlorobenzene, and chlorobenzene were studied using CuO/ SiO2 as a fly ash surrogate. These compounds served as model precursors that have been implicated in the formation of polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCDD/Fs). With FTIR, we determined that reactions of the model precursors with a substrate composed of CuO dispersed on silica result in the formation of a mixture of surface-bound phenolate and carboxylate partial oxidation products from 200 to 500 degrees C. Chemisorption of 2-chlorophenol and 1,2-dichlorobenzene resulted in the formation of identical surface-bound species. Using X-ray absorption near edge structure spectroscopy, we measured the time- and temperature-dependent reduction of Cu(II) in a fly ash surrogate during reaction with each precursor. It was demonstrated that CuI2O is the major reduction product in each case. The rate of Cu(II) reduction could be described using pseudo-first-order reaction kinetics with Arrhenius activation energies for reduction of Cu(II) of 112, 101, and 88 kJ mol(-1) for 2-chlorophenol, 1,2-dichlorobenzene, and chlorobenzene, respectively. We demonstrate that chlorinated phenol and chlorinated benzene both chemisorb to form chlorophenolate. Although chlorinated phenols chemisorb at a faster rate, chlorinated benzenes are found at much higher concentrations in incinerator effluents. The implication is that chlorinated benzenes may form 10 times more chlorophenolate precursors to PCDD/Fs than chlorinated phenols in combustion systems. PMID- 16245808 TI - Characterization of polycyclic aromatic hydrocarbons (PAHs) in different size fractions in deposited road particles (DRPs) from Lake Biwa area, Japan. AB - Deposited road particles (DRPs) collected from 13 heavily traveled roadways in an urban area of Southern Lake Biwa, Japan, were analyzed for polycyclic aromatic hydrocarbons (PAHs) in seven different particle size fractions (< or = 20 to 1000 2000 microm) and evaluated for the aryl hydrocarbon receptor (AhR) ligand activity by using a yeast bioassay. The mean compositions of individual PAHs to total PAH concentrations in different particle size fractions were 19-21% for pyrene, which was the most dominant component, 14-16% for fluoranthene, and 7-13% for benzo[g,h,i]perylene, which were the next dominant components. The total PAH distribution pattern in different particle size fractions of DRPs was different from the organic matter distribution pattern which increased with decreasing particle size of DRPs, and could be explained by the differences in their sources. Moreover, AhR ligand activities were observed in the DRP extracts of all size fractions. The activity of the DRP extracts from the smallest size fraction was approximately 5 times more potent than that of the largest size fraction in a yeast AhR ligand activity assay. The mean contribution (%) of benzo[k] fluoranthene, chrysene, benzo[b]fluoranthene, and benzo[a]pyrene to the AhR activity of DRP extracts in all size fractions was 1.43-4.11%, 1.63-3.53%, 0.63 1.69%, and 0.31-1.42%, respectively. Although the contribution of PAHs to AhR ligand activity presented in the DRP extracts was relatively low (below 10%), it may be best to remove these DRPs before discharge to receiving water environments. PMID- 16245809 TI - The role of resuspended soil in lead flows in the California South Coast Air Basin. AB - The inputs and outputs of airborne lead in the South Coast Air Basin of California (SOCAB) are quantified according to standard mass balance calculations. Results for 2001 show that approximately 49,000 kg of lead exitthe Basin each year, but traditional sources contribute only about 6500 kg of lead each year. We resolve this discrepancy through a simple computer model that quantifies the resuspension of lead-containing particles. Our results suggest that these lead particles were deposited during the years of leaded gasoline use and that resuspension is responsible for generating an additional 54,000 kg of airborne lead each year. This agrees roughly with estimated outputs. Thus, we conclude that resuspension, although an insignificant source of airborne lead during the era of leaded fuel, became a principal source in the SOCAB as lead emissions from vehicles declined. The results of the resuspension model further suggest that soil lead levels will remain elevated for many decades, in which case resuspension will remain a major source well into the future. PMID- 16245810 TI - Temporal and spatial trends of perfluorinated compounds in ringed seal (Phoca hispida) from Greenland. AB - Perfluorinated compounds (PFCs), such as perfluorooctane sulfonate (PFOS) and related compounds, have been identified as global pollutants and have shown their bioaccumulation into higher trophic levels in the food chain. PFCs have been found in remote areas far from sources, such as the Arctic. In this study spatial and temporal trends in the concentrations of selected PFCs were measured using archived liver samples of ringed seal (Phoca hispida) from East and West Greenland. The samples were collected in four different years at each location, between 1986 and 2003 in East Greenland and between 1982 and 2003 in West Greenland. PFOS was the major contributor to the burden of PFCs in samples, followed by perfluoroundecanoic acid (PFUnA). Perfluorononanoic acid (PFNA) and perfluorodecanoic acid (PFDA) were also detected in most samples. Perfluorohexane sulfonate (PFHxS) and perfluorooctane sulfonamide (PFOSA) were only found sporadically. Perfluorooctanoic acid was not found in detectable concentrations in any sample. Regression analysis of logarithmic transformed PFOS, PFDA, and PFUnA median concentrations indicated a significant temporal trend with increasing concentrations at both locations. A spatial trend in PFOS concentrations (ANOVA, p < 0.0001) was observed between the two sampling locations, with significantly higher concentrations in seals from East Greenland. PMID- 16245811 TI - Organophosphorus flame retardants and plasticizers in Swedish sewage treatment plants. AB - The levels, distribution, and possible sources of 12 organophosphorus flame retardants and plasticizers, some of which are reported to be toxic to aquatic organisms, were investigated in samples of influents, effluents, and sludge from 11 Swedish sewage treatment plants (STPs). The organophosphorus compounds (OPs) studied were poorly removed from the wastewater; especially the chlorinated OPs tended to pass through the STPs without being removed or degraded, while alkyl OPs, such as tributyl phosphate (TBP), were more successfully removed. In both influents and effluents, tris(2-butoxyethyl) phosphate and TBP were the most prominent substances followed by tris(2-chloroisopropyl) phosphate (TCPP). The highest concentration of any individual OP detected in the influents was 52 microg L(-1) (TBP). Ethylhexyl diphenyl phosphate and TCPP dominated in all sludge samples. A budget calculation comparing the annual amount of OPs in the influent received by Swedish STPs with the known amount of OPs imported indicated that approximately 15% is emitted to STPs. Of the total amount of OPs reaching the STPs annually, 49% is degraded, 50% (27 tons) is emitted to the recipients, and only 1% ends up in the sludge. The concentrations of most OPs were quite similar among the sampled STPs, indicating that the data may be applicable in other STPs. PMID- 16245812 TI - Molecular characteristics of urban organic aerosols from Nanjing: a case study of A mega-city in China. AB - Over 90 organic species have been determined in fine aerosols (PM2.5) collected during the summer and winter in Nanjing, a typical mega-city in China, using gas chromatography-mass spectrometry. The organic compounds detected were apportioned to four emission sources (i.e., plant emission, fossil fuel combustion, biomass burning, and soil resuspension) and secondary oxidation products. The most abundant classes of compounds are fatty acids, followed by sugars, dicarboxylic acids excluding oxalic and malonic acids, and n-alkanes, while alcohols, polyols/polyacids and lignin/sterols are less abundant. Total amounts of the seven classes of compounds were on average 938 ng m(-3) in the summer and 1301 ng m(-3) in the winter, respectively, contributing 0.26-1.96% of particle mass (PM2.5). In the summer, n-alkanes were heavily enhanced by vegetation emissions with a maximum carbon number (Cmax) at C29, whereas they were dominated by emissions from fossil fuels combustion with a Cmax at C22/ C23 in the winter. Concentrations of unsaturated fatty acids were lower in the summer than in the winter, being consistent with enhanced photooxidation of unsaturated fatty acids in the summer. Concentrations of dicarboxylic acids for the summer aerosols were much higher in the daytime than in the nighttime, indicating increased photochemical production in the daytime. In the summer, plant emissions were the most significant source of organic aerosols, contributing more than 33% of total compound mass (TCM), followed by fossil fuel combustion or secondary oxidation. In contrast, fossil fuel combustion was the dominant source of winter organic aerosols, contributing more than 51% of TCM, followed by plant emissions and secondary oxidation products. The quantitative results on sugars and lignin pyrolysis products further suggested that biomass burning and soil resuspension are also significant sources of urban organic aerosols. PMID- 16245813 TI - Perfluorinated alkyl substances in plasma, liver, brain, and eggs of glaucous gulls (Larus hyperboreus) from the Norwegian arctic. AB - Recent environmental surveys have ascertained the widespread occurrence of perfluorinated alkyl substances (PFAS) in tissues of wildlife from the Arctic. In the present study, we investigated the distribution of a suite of PFAS in plasma, liver, brain, and egg samples from adult glaucous gulls (Larus hyperboreus), an apex scavenger-predator seabird breeding in the Norwegian Arctic. Perfluorooctane sulfonate (PFOS) was the predominant PFAS in all samples and was present at concentrations that are the highest reported thus far in any arctic seabird species and populations. Among the body compartment/ tissue samples analyzed, PFOS was highest in plasma (48.1-349 ng/g wet weight (ww)), followed by liver approximately equal to egg > brain. Perfluorocarboxylic acids (PFCAs) with 8-15 carbon (C) atoms were found, with the highest concentrations determined in plasma (sum PFCA: 41.8-262 ng/g ww), whereas 5C- and 6C-PFCAs were below the limits of detection. Perfluorobutane sulfonate, perfluorooctane sulfonamide, and four saturated (8:2 FTCA and 10:2 FTCA) and unsaturated (8:2 FTUCA and 10:2 FTUCA) fluorotelomer carboxylic acids were not detected in any samples. Perfluorohexane sulfonate was measured at concentrations up to 2.71 ng/g ww. The accumulation profiles of PFCAs were characterized by high proportions of the long and odd numbered carbon-chain-length compounds, namely perfluoroundecanoic (11C) and perfluorotridecanoic acid (13C), although their individual contribution differed between the matrixes analyzed. Current PFAS concentrations suggest a bioaccumulation potential in Norwegian arctic glaucous gulls that needs to be assessed as part of a broad organohalogen contaminant cocktail with potential for mediating biological processes in this vulnerable top-predator marine species. PMID- 16245814 TI - Influence of dissolved organic matter from waste material on the phytotoxicity and environmental fate of triflusulfuron methyl. AB - Bioassays and chemical analyses were performed to study the effect of hydrophobic dissolved organic matter (HoDOM) extracted from a municipal waste compost (MWC) on the behavior of triflusulfuron methyl in soil and water. Bioassays with oilseed rape showed that HoDOM in culture solution lowered the effective dose 50 of triflusulfuron methyl by up to 4.8 times. Equilibrium dialysis experiments showed that in aqueous solution triflusulfuron methyl was adsorbed to HoDOM (K(OC) of 446.5 mL g(-1)). The half-life in water (pH 7.0) was increased from 52 to 76 days in the presence of HoDOM, but this cannot completely explain its phytotoxicity, as bioassays lasted for 21 days only. On the other hand, the addition of HoDOM to soils did not change the degradative behavior of triflusulfuron methyl. Fluorescein diacetate hydrolysis showed that HoDOM in soil did not significantly influence soil microbial activity, which may explain the above result on degradation in soil. Furthermore, in batch equilibrium experiments on soil, triflusulfuron methyl was only weakly adsorbed and the presence of HoDOM significantly modified the isotherm form. Results suggest that although the addition of exogenous HoDOM from MWC to soil did not influence the herbicide's persistence, its enhanced mobility could be of environmental concern and may deserve further research. PMID- 16245815 TI - Sorption of three tetracyclines by several soils: assessing the role of pH and cation exchange. AB - Tetracyclines (TCs) are widely used in veterinary medicine for treatment and prevention of disease and are present in animal waste products. Detection of TCs in soil, sediments, and water, and the growing concern of their potentially adverse effect on natural ecosystems have resulted in a need to understand their behavior in aqueous soil systems. TCs have multiple ionizable functional groups such that at environmentally relevant pH values, they may exist as a cation (+ 0 0), zwitterion (+ - 0), or a net negatively charged ion (+ - -), which complicates predicting their sorption, availability, and transport. We investigated the sorption of oxytetracycline (OTC), tetracycline (TC), and chlortetracycline (CTC) by several soils varying in pH, clay amount and type, cation exchange capacity (CEC), anion exchange capacity (AEC), and soil organic carbon in 0.01 N CaCl2, 0.001 N CaCl2, and 0.01 N KCI. All three TCs are highly sorbed, especially in acidic and high clay soils. When normalized to CEC, sorption tends to decrease with increasing pH. A sorption model in which species specific sorption coefficients normalized to pH-dependent CEC (Kd+00, kd+-0, and kd+--) and weighted by the pH-dependent fraction of each species fit the data well across all soils except for a soil rich in gibbsite and high in AEC. Resulting kd+00 values were more than an order of magnitude larger than values for either kd+0 and kd+--values such that kd+00 alone described most of the sorption observed as a function of pH for eight soils that varied in their mineralogy and pH (pH ranged from 4 to 8). PMID- 16245816 TI - Transformation of carbon tetrachloride by thiol reductants in the presence of quinone compounds. AB - Quinones are present in trace amounts in natural organic matter. The addition of thiol compounds to quinones produces reactive electron-transfer species that may be important for the transformation of chlorinated hydrocarbons under sulfate reducing conditions. This study systematically investigated the transformation of carbon tetrachloride (CCl4) in homogeneous aqueous solutions containing quinones as electron-transfer mediators and thiol compounds as bulk reductants. The thiol compounds, including sodium hydrosulfide (NaHS) and cysteine, were found to effectively transform CCl4. The transformation of CCl4 followed pseudo-first order kinetics, and the pseudo-first-order rate constants (kobs) were (3.24 +/- 0.46) x 10(-7) and 1.04 x 10(-7) s(-1), respectively, when solutions contained NaHS and cysteine alone. Addition of quinone compounds, including anthraquinone 2,6-disulfonate (AQDS), benzoquinone (BQ), juglone (JQ), naphthoquinone (NQ), lawsone (LQ), and menadione (MQ), increased the transformation rate and efficiency of CCl4. The kobs values for CCl4 transformation in the presence of quinones were 2.6-71 times higher than those for the thiol compounds alone. The enhancement efficiency followed the order JQ > NQ > BQ >> AQDS > LQ > MQ. Spectroscopic studies indicated that the quinone compounds generated various active electron-transfer mediators to transfer electrons from the bulk reductants to CCl4. BQ and NQ produced mercaptoquinones as active redox mediators that significantly enhanced the transformation rate of CCl4 in the presence of NaHS. The addition of thiol reductants produced large amounts of AQDS semiquinone radical as the electron shuttle. In addition, MQ and LQ were reduced by NaHS to give hydroquinone, which slightly enhanced the transformation efficiency of CCl4. These results clearly indicate that the enhanced efficiency of quinones for the transformation of chlorinated hydrocarbons is specifically related to the produced reactive species. Mercaptoquinone is a more active mediator than either semiquinone or hydroquinone for transferring electrons in a reducing environment containing thiol reductants. PMID- 16245817 TI - Carbamazepine and its metabolites in wastewater and in biosolids in a municipal wastewater treatment plant. AB - Pharmaceutically active compounds (PhACs) are discharged into the environment from domestic wastewater treatment plants (WWTPs). In this study, we determined the distribution of the anti-epileptic drug, carbamazepine (CBZ), and its major metabolites and caffeine in both aqueous and solid phases through different treatment processes of a WWTP. A method was developed to extract samples of biosolids using pressurized liquid extraction (PLE), coupled with cleanup of extracts using solid-phase extraction. Samples of biosolids and wastewater were analyzed for caffeine and CBZ and five of its metabolites, 10,11-dihydro-10,11 epoxycarbamazepine (CBZ-EP), 11-dihydro-10,11-epoxycarbamazepine (CBZ-DiOH), 2 hydroxycarbamazepine (CBZ-20H), 3-hydroxycarbamazepine (CBZ-30H), and 10,11 dihydro-10-hydroxycarbamazepine (CBZ-100H). The analytes were quantified using liquid chromatography-electrospray ionization tandem mass spectrometry. The recoveries of the analytes were 82.1-91.3% from raw biosolids and 80.1-92.4% from treated biosolids, and the limits of detection were 0.06-0.50 and 0.06-0.40 microg/kg on a wet weight basis for raw and treated biosolids, respectively. The behavior of carbamazepine and its metabolites, together with caffeine as a marker of domestic inputs, was investigated in the WWTP for the City of Peterborough, ON, Canada, which utilizes secondary sewage treatment technologies. CBZ, CBZ-2OH, CBZ-30H, and CBZ-DiOH were detected at concentrations of 69.6, 1.9, 1.6, and 7.5 microg/kg (dry weight), respectively, in untreated biosolids and at concentrations of 258.1, 3.4, 4.3, and 15.4 microg/kg (dry weight), respectively, in treated biosolids. However, CBZ-EP and CBZ-100H were not detected in any of the biosolid samples. CBZ and its five metabolites were detected in all wastewater samples collected from four different stages of treatment. The results showed that 29% of the carbamazepine was removed from the aqueous phase during treatment in the WWTP, while the metabolites were not effectively removed. Concentrations of caffeine were reduced by 99.9% in the aqueous phase, which appeared to be due primarily to degradation. Caffeine was also detected at concentrations of 165.8 and 7.6 microg/kg (dry weight) in raw and treated biosolids, respectively. Because of differences in hydrophobicity, CBZ is the primary analyte in biosolids, while CBZ-DiOH is the primary analyte in the aqueous phase of the wastewater. A mass balance calculation showed that the majority of CBZ and its metabolites exist in the aqueous phase (i.e., wastewater), ratherthan in the biosolids, 78 g of CBZ and its metabolites enters the Peterborough WWTP daily, and 91 g is discharged from the WWTP daily in the combined suspended solids and aqueous phases of the wastewater. The calculated daily inputs into the WWTP are somewhat less than the inputs of 192 g estimated from Canadian annual sales data for CBZ. PMID- 16245818 TI - An isotope exchange technique to assess mechanisms of sorption hysteresis applied to naphthalene in kerogenous organic matter. AB - The sorption of organic compounds to natural sorbents is often found to show hysteresis. The objective of this study was to develop an experimental technique based on the use of 14C isotopes to distinguish hysteresis due to experimental artifacts from true hysteresis due to thermodynamically irreversible processes. The study was also designed to investigate causation of true hysteresis (irreversible sorption). The technique determines the rates and the degree of isotope exchange (IE) on equilibrated sorption and desorption points at different constant bulk chemical concentrations. The technique was applied to the sorption of naphthalene (NAPH) on Beulah-Zap lignite, a low rank reference coal composed mainly of kerogen. Sorption of bulk was found to be reversible below 10(-5) g L( 1), but irreversible above 10(-4) g L(-1). Complete isotope exchange on sorption and desorption points that defined an irreversible cycle demonstrated that hysteresis was true. A comparison of normalized uptake and release kinetics of labeled and bulk NAPH at different concentrations revealed slow structural deformation processes of the sorbent during bulk sorption and desorption. This is taken as corroborating evidence for the pore deformation hypothesis of hysteresis in which incoming sorbate molecules induce quasi-reversible changes in the organic matter that lead to different pathways for sorption and desorption. Although unable to rule it out completely, the data demonstrate that physical entrapment of sorbate molecules plays a minor, if any, role to the observed hysteresis in this system. PMID- 16245819 TI - Phosphorus speciation in manure and manure-amended soils using XANES spectroscopy. AB - Previous studies suggested an increase in the proportion of calcium phosphates (CaP) of the total phosphorus (P) pool in soils with a long-term poultry manure application history versus those with no or limited application histories. To understand and predict long-term P accumulation and release dynamics in these highly amended soils, it is important to understand what specific P species are being formed. We assessed forms of CaP formed in poultry manure and originally acidic soil in response to different lengths of mostly poultry manure applications using P K-edge X-ray absorption near-edge structure (XANES) spectroscopy. Phosphorus K-edge XANES spectra of poultry manure showed no evidences of crystalline P minerals but dominance of soluble CaP species and free and weakly bound phosphates (aqueous phosphate and phosphate adsorbed on soil minerals). Phosphate in an unamended neighboring forest soil (pH 4.3) was mainly associated with iron (Fe) compounds such as strengite and Fe-oxides. Soils with a short-term manure history contained both Fe-associated phosphates and soluble CaP species such as dibasic calcium phosphate (DCP) and amorphous calcium phosphate (ACP). Long-term manure application resulted in a dominance of CaP forms confirming our earlier results obtained with sequential extractions, and a transformation from soluble to more stable CaP species such as beta-tricalcium calcium phosphate (TCP). Even after long-term manure application (> 25 yr and total P in soil up to 13,307 mg kg(-1)), however, none of the manure-amended soils showed the presence of crystalline CaP. With a reduction or elimination of poultry manure application to naturally acidic soils, the pH of the soil is likely to decrease, thereby increasing the solubility of Ca-bonded inorganic P minerals. Maintaining a high pH is therefore an important strategy to minimize P leaching in these soils. PMID- 16245820 TI - Voltammetry of copper sulfide particles and nanoparticles: investigation of the cluster hypothesis. AB - An association of Cu with sulfide in aerobic natural waters has been attributed to these components' coexistence in clusters of sizes intermediate between mononuclear complexes and colloidal particles. This hypothesis is investigated here. Copper sulfide solid phases display size-related voltammetric behavior at Hg electrodes. Suspensions of copper sulfide powders held at accumulation potentials of 0 to -0.2 V (vs Ag/AgCl) produce voltammetric peaks near -0.15, 0.65, and -0.95 V during subsequent cathodic scans. The first two peaks arise from electrochemically generated Cu-oxyhydroxides and HgS; the -0.95 V peak arises from reduction of sorbed copper sulfide particles. Nanoparticles of radius approximately 10(-8) m produce the third peak even without stirring or accumulation. Still smaller analytes give only the first two peaks. Published evidence alleging production of subnanometer copper sulfide clusters during titrations of Cu2+ and HS- was not reproduced when sulfide oxidation was avoided. Instead, such titrations apparently generate nanoparticles. The titration stoichiometry is 1/1, consistent with previous descriptions of this process: Cu2+ + HS- --> 1/2Cu2S x S0 (brown sol) --> CuS (green sol). Titrating Cu2+ into organic-rich (muscilaginous) Adriatic Sea water, which contains 10(-7) M natural thiols and sulfide, produces solid products. In the future, voltammetry might prove useful for studying semiconductor sulfide nanoparticles in nature. PMID- 16245821 TI - Study on the structure and mutagenicity of a new disinfection byproduct in chlorinated drinking water. AB - The structure of a new chlorinated disinfection byproduct (DBP) in drinking water is studied. The new DBP was prepared through chlorinating its best precursor, and it was extracted and isolated from the chlorination products. Its structure is identified as 2,2,4-trichloro-5-methoxycyclopent-4-ene-1,3-dione (TCMCD) through FTIR, MS, 1H NMR, and 13C NMR spectroscopic analysis and single-crystal X-ray diffraction method. Results of the Ames test with tester strain TA100 indicated that the newly identified DBP TCMCD is a mutagen. PMID- 16245822 TI - Primary consumer stable nitrogen isotopes as indicators of nutrient source. AB - Non-point source loading of nitrogen and phosphorus is a primary cause of eutrophication of inland waters, although the diffuse and variable nature of nutrient inputs makes it difficult to trace and identify nutrient pathways. Stable nitrogen isotope values (delta15N) in aquatic biota are thought to reflect anthropogenic nutrient inputs, and they may be a promising tool fortracing nutrient sources in watersheds. We measured delta15N of aquatic consumers from a suite of 27 Danish lakes spanning a range of trophic states (oligotrophic to eutrophic) and land uses (forest, urban, agriculture). Primary consumer delta15N values (PCdelta15N) varied more than 14% among lakes. Models of PCdelta15N were developed from limnological, nitrogen loading, and nitrogen source variables using an information-theoretic approach (Akaike's Information Criteria, AIC). Models based on land use/ land cover performed best, indicating that elevated delta15N is not only associated with high nitrogen loading, but is also reflective of nitrogen source. Urban and agricultural land use in the watershed, and particularly within the riparian buffer areas, was quantitatively linked to elevated biotic delta15N. PMID- 16245823 TI - Fluorotelomer alcohol biodegradation-direct evidence that perfluorinated carbon chains breakdown. AB - There is increasing scientific interest to understand the environmental fate of fluorotelomer alcohols (FTOHs) and fluorotelomer-based products which may break down to FTOHs. Both are expected to enter aqueous waste streams, which would be processed in a wastewater treatment plant and therein subject to microbial biodegradation. We investigated the biodegradation of 3-14C, 1H,1H,2H,2H perfluorodecanol [CF3(CF2)6(14)CF2CH2CH2OH, 14C-8-2 FTOH] in mixed bacterial culture and activated sludge. 14CO2 and 14C-organic volatiles in the headspace of the sealed bottles and bottles with continuous air flow were analyzed up to 4 months. After sample extraction with acetonitrile, 14C-labeled biotransformation products (metabolites) were quantified by LC/ARC (on-line liquid chromatography/ accurate radioisotope counting) and identified by quadrupole time-of-flight (Q TOF) mass spectrometry and GC/MSD (mass selective detector). Three metabolites not yet reported in the literature have been identified as CF3(CF2)6(14)CHOHCH3 (7-2 sFTOH), CF3(CF2)6(14)CH=CHCOOH (7-3 unsaturated acid or 7-3 u acid), and CF3(CF2)6(14)CH=CHCONH2 (7-3 u amide) along with five previously reported metabolites [CF3(CF2)6(14)CF2CH2CHO (8-2 FTAL), CF3(CF2)6 (14)CF2CH2COOH (8-2 acid), CF3(CF2)6(14)CF=CHCOOH (8-2 u acid), CF3(CF2)6(14)CH2CH2COOH (7-3 acid), and CF3(CF2)6(14)COOH (PFOA)]. No CF3(CF2)6(14)CF2COOH (14C-PFNA) was observed, indicating that alpha-oxidation does not take place. It was found that strong adsorption to the activated sludge and subsequent transformation, even under continuous air flow, greatly reduced partitioning of 8-2 FTOH or any transformation products to air. CF3(CF2)4COOH (PFHA; perfluorohexanoic acid) was observed and increased in mixed bacterial culture over 28 days and accounted for about 1% of the initial 14C-8-2 FTOH concentration from day 28 to day 90. 14CO2 accounted for 1% of initial 14C in activated sludge with continuous air flow at day 1 and increased over time. In closed bottles, 14CO2 in the headspace of activated sludge medium increased to 12% of the available 14C over 135 days with periodic addition of ethanol, as compared to 3% when no additional ethanol was added. These results show that replenishment of organic carbon enhanced microbial mineralization of multiple--CF2--groups in the fluorocarbon chain of 14C-8-2 FTOH. At day 90 the net increase of fluoride ion in the mixed bacterial culture was 93 microg L(-1), equivalent to 12% of total mineralization (destruction) of the 14C-8-2 FTOH. These results demonstrate that perfluorinated carbon bonds of 14C-8-2 FTOH are defluorinated and mineralized by microorganisms under conditions which may occur in a wastewater treatment plant, forming shorter fluorinated carbon metabolites. PMID- 16245824 TI - Comparative sorption and desorption of benzo[a]pyrene and 3,4,3',4' tetrachlorobiphenyl in natural lake water containing dissolved organic matter. AB - The sorption and desorption of two model compounds, benzo[a]pyrene (BaP) and 3,4,3',4'-tetrachlorobiphenyl (TCBP), were studied in natural lake water with high dissolved organic matter (DOM) content using the equilibrium dialysis and Tenax extraction methods. The sorption of TCBP was lower and reached steady value more slowly than did BaP. Tenax extraction revealed at least two differently desorbing fractions for both model compounds, which also supported the conclusion that DOM-HOC associations may involve several mechanisms. The rapidly desorbing fraction may be attributed to freely dissolved and loosely sorbed compound, whereas the more strongly sorbed fraction may indicate the presence of specific binding sites. The data indicated that the association between hydrophobic organic contaminants (HOC) and DOM is not simply absorption that is solely driven by the lipophilicity of the sorbates. Although contact time had a rather negligible effect on the sorption of BaP, the proportion of desorption resistant fraction increased with time, whereas the desorption of TCBP was less affected by contact time. Steric factors may be the cause of the lower sorption and smaller desorption resistant fraction of TCBP. The results indicate potential differences in the behavior of PAHs and PCBs in the aquatic environment. PMID- 16245825 TI - Solubility and sorption by soils of 8:2 fluorotelomer alcohol in water and cosolvent systems. AB - The solubility and sorption by five soils of 8:2 fluorotelomer alcohol (FTOH) were measured from water and cosolvent/ water solutions. Aqueous solubility and soil-water distribution coefficients (Kd,w, L kg(-1)) were extrapolated from cosolvent data using a log-linear cosolvency model and compared to direct aqueous measurements. Liquid chromatography tandem mass spectrometry with electrospray ionization was employed to analyze the 8:2 FTOH in solutions and soil extracts. The cosolvent-extrapolated water solubility is 0.224 mg L(-1), in good agreement with the measured value of 0.194 mg L(-1). All sorption isotherms were generally linear regardless of cosolvent composition or soil organic carbon (OC) content. Kd,w values extrapolated from cosolvent data were similar but consistently higher than those measured in aqueous solutions. The latter was hypothesized to be due to dissolved OC (DOC) in the aqueous slurries. An average log KDOC of 5.30 was estimated and supported by DOC and Kd,w measurements at two soil-water ratios. Sorption appeared to be driven by hydrophobic partitioning with a log KOC value of 4.13 +/- 0.16. Irreversible sorption was also observed and appeared to be related to OC content, with the extraction efficiency reduced from 85% to 45% with increasing contact time from 3 to 72 h for the highest OC soil. PMID- 16245826 TI - Biofilm formation characteristics of bacterial isolates retrieved from a reverse osmosis membrane. AB - High-quality water purification systems using reverse osmosis (RO) membrane separation have faced a major challenge related to biofilm formation on the membrane surface, or biofouling. To understand this issue, the biofilm formation characteristics of four bacterial isolates previously retrieved from an RO membrane treating potable water were investigated. Biofilm formation of all four isolates occurred to different extents in microtiter plates and could be related to one or more cell properties (hydrophobicity, surface charge, and motility). For Dermacoccus sp. strain RO12 and Microbacterium sp. strain RO18, bacterial adhesion was facilitated by cell surface hydrophobicity, and for Rhodopseudomonas sp. strain RO3, adhesion was assisted by its low surface charge. Sphingomonas sp. strain RO2 possessed both twitching and swarming motilities, which could be important in mediating surface colonization. Further, strains RO2, RO3, and RO12 did not exhibit swimming motility, suggesting that they could be transported to RO membrane surfaces by other mechanisms such as convective permeate flow. The biofilm formation of RO2 was further tested on different RO membranes made of cellulose acetate, polyamide, and thin film composite in continuous flow cell systems. The resultant RO2 biofilms were independent of membrane surface properties and this was probably related to the ex-opolysaccharides secreted bythe biofilm cells. These results suggested that RO2 could colonize RO membranes effectively and could be a potential fouling organism in RO membranes for freshwater purification. PMID- 16245827 TI - A multivariate statistical approach to spatial representation of groundwater contamination using hydrochemistry and microbial community profiles. AB - Managers of landfill sites are faced with enormous challenges when attempting to detect and delineate leachate plumes with a limited number of monitoring wells, assess spatial and temporal trends for hundreds of contaminants, and design long term monitoring (LTM) strategies. Subsurface microbial ecology is a unique source of data that has been historically underutilized in LTM groundwater designs. This paper provides a methodology for utilizing qualitative and quantitative information (specifically, multiple water quality measurements and genome-based data) from a landfill leachate contaminated aquifer in Banisveld, The Netherlands, to improve the estimation of parameters of concern. We used a principal component analysis (PCA) to reduce nonindependent hydrochemistry data, Bacteria and Archaea community profiles from 16S rDNA denaturing gradient gel electrophoresis (DGGE), into six statistically independent variables, representing the majority of the original dataset variances. The PCA scores grouped samples based on the degree or class of contamination and were similar over considerable horizontal distances. Incorporation of the principal component scores with traditional subsurface information using cokriging improved the understanding of the contaminated area by reducing error variances and increasing detection efficiency. Combining these multiple types of data (e.g., genome-based information, hydrochemistry, borings) may be extremely useful at landfill or other LTM sites for designing cost-effective strategies to detect and monitor contaminants. PMID- 16245828 TI - Estuarial fingerprinting through multidimensional fluorescence and multivariate analysis. AB - As part of a strategy for preventing the introduction of aquatic nuisance species (ANS) to U.S. estuaries, ballast water exchange (BWE) regulations have been imposed. Enforcing these regulations requires a reliable method for determining the port of origin of water in the ballast tanks of ships entering U.S. waters. This study shows that a three-dimensional fluorescence fingerprinting technique, excitation emission matrix (EEM) spectroscopy, holds great promise as a ballast water analysis tool. In our technique, EEMs are analyzed by multivariate classification and curve resolution methods, such as N-way partial least squares Regression-discriminant analysis (NPLS-DA) and parallel factor analysis (PARAFAC). We demonstrate that classification techniques can be used to discriminate among sampling sites less than 10 miles apart, encompassing Boston Harbor and two tributaries in the Mystic River Watershed. To our knowledge, this work is the first to use multivariate analysis to classify water as to location of origin. Furthermore, it is shown that curve resolution can show seasonal features within the multidimensional fluorescence data sets, which correlate with difficulty in classification. PMID- 16245829 TI - Immunological impact of metals in harbor seals (Phoca vitulina) of the North Sea. AB - Environmental pollutants may affect the immune system of marine mammals in many areas of the industrialized world. This study provides the first evidence for metal-induced hypersensitivity in harbor seals and demonstrates a relationship between this immunopathy and the level of metals in blood. The concentrations of 20 essential and nonessential elements were analyzed in the blood of 13 harbor seals from the North Sea. In addition, their T-lymphocyte response to metals in terms of hypersensitivity was investigated using a lymphocyte transformation test (LTT) according to the MELISA (memory lymphocyte immuno-stimulation assay) modification. The results showed metal hypersensitivities in 7 of 11 seals investigated in MELISA (data from two seals could not be assessed), reflecting a positive or possible positive reaction in 13 of 154 total single tests. Four animals responded to one metal and three animals to multiple metals. The sensitizing metals were molybdenum (Mo), titanium (Ti), nickel (Ni), chromium (Cr), aluminum (Al), lead (Pb), and tin (Sn). Furthermore, the seals with a Ni-, Al-,.and Cr-sensibilization showed the highest concentrations of these metals in blood. In 8 of the 13 positive cases, elevated blood metal concentrations correlated with the hypersensitivity reaction. Summarizing, we demonstrate in this first pilot study the potential immunological impact of metals in seals, a topic rarely investigated previously. Our results show the value of a combined biological and effect-monitoring tool to investigate pollution-induced immunopathies in live animals. PMID- 16245830 TI - Mechanistic aspects of pyrite oxidation in an oxidizing gaseous environment: an in situ HATR-IR isotope study. AB - The reaction of FeS2 (pyrite) with gaseous H2O, O2, and H2O/O2 was investigated using horizontal attenuated total reflection Fourier transform infrared spectroscopy (HATR-FTIR). Spectra were interpreted with the aid of hybrid molecular orbital/density functional theory calculations of sulfate-iron hydroxide clusters. Reaction of pyrite in gaseous H2O led primarily to the formation of iron hydroxide on pyrite. Exposure of the pyrite to gaseous O2 after exposure to H2O vapor led to the formation of sulfur oxyanions that included SO42 . Isotopic labeling experiments showed that after this exposure sequence the oxygen in the sulfate product was primarily derived from the H2O reactant. If, however, pyrite was exposed to gaseous O2 prior to pure H2O vapor, both SO42- and iron oxyhydroxide became significant products. Isotopic rabeling experiments using the O2-then-H2O sequence showed that the oxygen in the SO42- product was derived from both H2O and O2. The results indicate that H2O and O2 exhibit a competitive adsorption on pyrite, with H2O blocking surface sites for O2 adsorption. The extent of oxygen incorporation from either the H2O or the O2 component into the surface-bound sulfur oxyanion product appears to be a strong function of the relative concentration ratio of the reactant H2O and O2. PMID- 16245831 TI - Assessing sequestration of selected polycyclic aromatic hydrocarbons by use of adsorption modeling and temperature-programmed desorption. AB - Sequestration of phenanthrene and pyrene was investigated in two soils--a sandy soil designated SBS and a silt-loam designated LHS--by combining long-term batch sorption studies with thermal desorption and pyrolysis of amended soil samples. The Polanyi-based adsorption volume and the adsorbed solute mass increased with aging for both soils, thus demonstrating the mechanism for observed sequestration. Despite rigorous thermal analysis, 30-62% (SBS sand) and 8-30% (LHS silt-loam) of phenanthrene could not be recovered after 30-270 days of sorption, with the increase in desorption resistance showing greater significance in SBS sand. For both soils, these values were 20-65% of adsorbed phenanthrene mass. Activation energies estimated from the temperature-programmed desorption (TPD) of sorbed phenanthrene at < or = 375 degrees C were 51-53 kJ/mol, consistent with values derived for desorption of organic compounds from humic materials. The activated first-order model fitting of observed TPD data supports the conclusion that the desorption-resistant fraction of phenanthrene has become sequestered onto condensed organic domains and requires temperatures exceeding 600 degrees C to be released. The work demonstrates the use of thermal analysis in complementing the Polanyi-based adsorption modeling approach for assessing the mechanistic basis for sequestration of organic contaminants in soils. PMID- 16245832 TI - Geophysical imaging of stimulated microbial biomineralization. AB - Understanding how microorganisms influence the physical and chemical properties of the subsurface is hindered by our inability to observe microbial dynamics in real time and with high spatial resolution. Here, we investigate the use of noninvasive geophysical methods to monitor biomineralization at the laboratory scale during stimulated sulfate reduction under dynamic flow conditions. Alterations in sediment characteristics resulting from microbe-mediated sulfide mineral precipitation were concomitant with changes in complex resistivity and acoustic wave propagation signatures. The sequestration of zinc and iron in insoluble sulfides led to alterations in the ability of the pore fluid to conduct electrical charge and of the saturated sediments to dissipate acoustic energy. These changes resulted directly from the nucleation, growth, and development of nanoparticulate precipitates along grain surfaces and within the pore space. Scanning and transmission electron microscopy (SEM and TEM) confirmed the sulfides to be associated with cell surfaces, with precipitates ranging from aggregates of individual 3-5 nm nanocrystals to larger assemblages of up to 10-20 microm in diameter. Anomalies in the geophysical data reflected the distribution of mineral precipitates and biomass over space and time, with temporal variations in the signals corresponding to changes in the aggregation state of the nanocrystalline sulfides. These results suggest the potential for using geophysical techniques to image certain subsurface biogeochemical processes, such as those accompanying the bioremediation of metal-contaminated aquifers. PMID- 16245834 TI - A novel method for determination of size resolved, submicrometer particle traffic emission factors. AB - A novel approach to determine size-segregated particle number emission factors for traffic is presented. It was proven that using limited data sets (800-2000 samples) statistically significant emission factors from road traffic can be extracted. In this study data from four sites were used for calculating emission factors (rural and urban roadside, urban rooftop, and urban background). The measurements were performed using SMPS/DMPS (scanning or differential particle sizers) from TSI and commercial gas analyzers. Describing the particle concentration as a ratio to an exhaust trace gas, e.g. NOx, the dilution effect will be minimized. This ratio is easily compared among different studies. By knowledge of the emission factor of the chosen trace gas the emission ratio can be converted to an emission factor for particle numbers of defined particle sizes. For the presented method only one measurement site is needed, where the difference between high and low (background) traffic exposure is used. To define high and low traffic exposure, the best result was obtained using high ratio of [NO] to [NO2] and low [NOx], respectively. Emission ratios for 10-100-nm particles at two road sites, one high-speed 90-kmph rural case and one urban, slower, and more congested situation, were determined to (35 +/- 15) x 10(14) and (24 +/- 8) x 10(14) particles per mole NOx, respectively. PMID- 16245833 TI - Assessing organic contaminants in fish: comparison of a nonlethal tissue sampling technique to mobile and stationary passive sampling devices. AB - As concerns mount over the human health risks associated with consumption of fish contaminated with persistent organic pollutants, there exists a need to better evaluate fish body burdens without lethally sampling many of the important commercial and sport species of interest. The aim of this study was to investigate two novel methods for estimating organic contaminants in fish that are a concern for both fish and human health. The removal of fish adipose fins, commonly done in mark-recapture studies with salmonid species, was evaluated as a nonlethal sampling technique to estimate concentrations of polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs) in flathead catfish (Pylodictis olivaris), relative to those found in muscle fillets of the same fish. We also assessed the efficacy of using poly(dimethylsiloxane) (PDMS) as a mobile passive sampling device (PSD) attached directly to wild flathead catfish for assessing location-specific exposure of the fish to waterborne contaminants. The results of this study have demonstrated for the first time that organic contaminant concentrations in adipose fin were highly correlated (R2 = 0.87) with muscle fillet concentrations, indicating that the adipose fin of certain fishes may be used to accurately estimate tissue concentrations without the need for lethal sampling. Moreover, mobile PSDs attached directly to fish and used here for the first time accurately estimated ultratrace concentrations of waterborne PCBs and OCPs without any apparent harm to the fish, indicating that there are no practical or physical barriers to the use of mobile passive samplers attached to aquatic organisms. Among the many practical implications of this research, two potential priority items include the analysis of organic contaminants in farm raised and sport fish intended for human consumption, without the economic and population losses associated with lethally sampling fish to obtain tissues, and identifying specific areas where fish may be accumulating large portions of their contaminant burden. PMID- 16245835 TI - Simultaneous determination of mono- and dicarboxylic acids, omega-Oxo-carboxylic acids, midchain ketocarboxylic acids, and aldehydes in atmospheric aerosol samples. AB - This paper describes a method for the simultaneous determination of monocarboxylic acids (C6-C34), dicarboxylic acids (C2-C24), omega-oxo-carboxylic acids (C2-C9), ketocarboxylic acids (pyruvic and pinonic acid), and select aldehydes (glyoxal, methylglyoxal, and nonanal) in atmospheric particles. Quantification of these compounds gives information on important chemical characteristics of aerosols for source apportioning of aerosol organics and for studying atmospheric processes leading to secondary organic aerosol formation. These target analytes were determined as their butyl ester or butyl acetal derivatives using gas-chromatography mass spectrometry. The method is modified from a method described by Kawamura. Kawamura's original method involved a water extraction step, which practically restricted the method to the determination of only those compounds that are water-soluble. Our method eliminates the water extraction step and combines extraction and derivatization of the target compounds in one step. A mixture of hexane/butanol/BF3 simultaneously derivatizes the polar function groups (i.e., -COOH, -C=O) and extracts the target analytes from the aerosol filter substrates. A prominent advantage of our method is improved recoveries for the more volatile analytes in the target compound classes as a result of eliminating the water evaporation step. Recoveries better than 66% were obtained for the target analytes, including the relatively volatile ones. This improvement for the light species has allowed detection of a new midchain ketocarboxylic acid, 4-oxopentanoic acid, which would have escaped detection by the Kawamura method because of its high susceptibility to evaporative loss. Examples are presented to demonstrate the use of this method in analysis of ambient aerosol samples. PMID- 16245836 TI - Bacterial bioassay for rapid and accurate analysis of arsenic in highly variable groundwater samples. AB - In this study, we report the first ever large-scale environmental validation of a microbial reporter-based test to measure arsenic concentrations in natural water resources. A bioluminescence-producing arsenic-inducible bacterium based on Escherichia coli was used as the reporter organism. Specific protocols were developed with the goal to avoid the negative influence of iron in groundwater on arsenic availability to the bioreporter cells. A total of 194 groundwater samples were collected in the Red River and Mekong River Delta regions of Vietnam and were analyzed both by atomic absorption spectroscopy (AAS) and by the arsenic bioreporter protocol. The bacterial cells performed well at and above arsenic concentrations in groundwater of 7 microg/L, with an almost linearly proportional increase of the bioluminescence signal between 10 and 100 microg As/L (r2 = 0.997). Comparisons between AAS and arsenic bioreporter determinations gave an overall average of 8.0% false negative and 2.4% false positive identifications for the bioreporter prediction at the WHO recommended acceptable arsenic concentration of 10 microg/L, which is far betterthan the performance of chemical field test kits. Because of the ease of the measurement protocol and the low application cost, the microbiological arsenic test has a great potential in large screening campaigns in Asia and in other areas suffering from arsenic pollution in groundwater resources. PMID- 16245837 TI - Atmospheric size distribution of PAHs: evidence of a high-volume sampling artifact. AB - The purpose of this study was to characterize atmospheric levels of four- to six ring polycyclic aromatic hydrocarbons (PAHs) in the vicinity of a horizontal stud Soderberg aluminum smelter in terms of the size distribution of particulate matter (September to December 2002). It was found that the vast majority of the PAHs was associated with particle diameters less than 1 and 3 microm. A profile comparison of the PAH mixture--using benzo[a]pyrene (B[a]P) relative abundance ratios (PAH/B[a]P)--for the cascade impactor filters indicated the formation of a sampling artifact. Overall, the PAH stability scale generated in this study agrees with those produced experimentally for ozone and nitrogen dioxides or developed using other in situ measurement techniques. Correlations of the four- to six-ring PAHs with other atmospheric variables suggested that smelter plume conditions and particle characteristics may play a potentially important role in the overall PAH reactivity. To our knowledge, this is the first study to report a sampling artifact for the four- to six-ring PAHs during in situ high volume sampling under real world conditions. PMID- 16245838 TI - Emissions of toxic pollutants from compressed natural gas and low sulfur diesel fueled heavy-duty transit buses tested over multiple driving cycles. AB - The number of heavy-duty vehicles using alternative fuels such as compressed natural gas (CNG) and new low-sulfur diesel fuel formulations and equipped with after-treatment devices are projected to increase. However, few peer-reviewed studies have characterized the emissions of particulate matter (PM) and other toxic compounds from these vehicles. In this study, chemical and biological analyses were used to characterize the identifiable toxic air pollutants emitted from both CNG and low-sulfur-diesel-fueled heavy-duty transit buses tested on a chassis dynamometer over three transient driving cycles and a steady-state cruise condition. The CNG bus had no after-treatment, and the diesel bus was tested first equipped with an oxidation catalyst (OC) and then with a catalyzed diesel particulate filter (DPF). Emissions were analyzed for PM, volatile organic compounds (VOCs; determined on-site), polycyclic aromatic hydrocarbons (PAHs), and mutagenic activity. The 2000 model year CNG-fueled vehicle had the highest emissions of 1,3-butadiene, benzene, and carbonyls (e.g., formaldehyde) of the three vehicle configurations tested in this study. The 1998 model year diesel bus equipped with an OC and fueled with low-sulfur diesel had the highest emission rates of PM and PAHs. The highest specific mutagenic activities (revertants/microg PM, or potency) and the highest mutagen emission rates (revertants/mi) were from the CNG bus in strain TA98 tested over the New York Bus (NYB) driving cycle. The 1998 model year diesel bus with DPF had the lowest VOCs, PAH, and mutagenic activity emission. In general, the NYB driving cycle had the highest emission rates (g/mi), and the Urban Dynamometer Driving Schedule (UDDS) had the lowest emission rates for all toxics tested over the three transient test cycles investigated. Also, transient emissions were, in general, higher than steady-state emissions. The emissions of toxic compounds from an in-use CNG transit bus (without an oxidation catalyst) and from a vehicle fueled with low sulfur diesel fuel (equipped with DPF) were lower than from the low-sulfur diesel fueled vehicle equipped with OC. All vehicle configurations had generally lower emissions of toxics than an uncontrolled diesel engine. Tunnel backgrounds (measurements without the vehicle running) were measured throughout this study and were helpful in determining the incremental increase in pollutant emissions. Also, the on-site determination of VOCs, especially 1,3-butadiene, helped minimize measurement losses due to sample degradation after collection. PMID- 16245839 TI - Compost-based permeable reactive barriers for the source treatment of arsenic contaminations in aquifers: column studies and solid-phase investigations. AB - The bulk of arsenic (As) at contaminated sites is frequently associated with iron (hydr)oxides. Various studies ascribe increasing dissolved As concentrations to the transformation of iron (hydr)oxides into iron sulfides, which is initiated by dissolved sulfide. We investigated whetherthis processes can be utilized as a source treatment approach using compost-based permeable reactive barriers (PRB), which promote microbial sulfate reduction. Arsenic-bearing aquifer sedimentfrom a contaminated industrial site showed a decrease in As content of <10% after 420 days of percolation with sulfide-free artificial groundwater. In contrast, water that had previously passed through organic matter and exhibited sulfide concentrations of 10-30 mg/L decreased As content in the sediment by 87% within 360 days. X-ray diffraction showed no arsenic sulfides, but XANES spectra (X-ray absorption near edge structure) and associated linear combinations revealed that adsorbed arsenate of the original sediment was in part reduced to arsenite and indicated the formation of minor amounts of a substance that contains As and sulfur. The speciation of dissolved As changed from initially As(V)-dominated to As(III)-dominated after sulfide flushing was started, which increases the mobility of As. Because sulfide can be supplied not only by compost-based PRBs but also by direct injection, sulfide flushing has a wide range of application for the source treatment of arsenic. PMID- 16245840 TI - Fabrication of catalytic membranes for the treatment of drinking water using combined ozonation and ultrafiltration. AB - The removal of disinfection byproducts and their precursors was investigated using a combined ozonation-ultrafiltration system. A commercial membrane was coated 20 or 40 times with iron oxide nanoparticles (4-6 nm in diameter). With this membrane, the concentration of dissolved organic carbon was reduced by >85% and the concentrations of simulated distribution system total trihalomethanes and simulated distribution system halo acetic acids decreased by up to 90% and 85%, respectively. When the coated membrane was used, the concentrations of aldehydes, ketones, and ketoacids in the permeate were reduced by >50% as compared to that obtained with the uncoated membranes. Hydroxyl or other radicals produced at the iron oxide coated membrane surface as a result of ozone decomposition are believed to have enhanced the degradation of the natural organic matter, thereby reducing the concentration of disinfection byproducts. While increasing the number of times the membrane was coated from 20 to 40 did not significantly reduce the concentrations of most of the parameters measured, it did result in a significant decrease in the concentrations of ozonation byproducts. Increasing the sintering temperature from 500 to 900 degrees C also resulted in an improvement in the removal of the ozonation byproducts. PMID- 16245841 TI - Removal of methylated arsenic in groundwater with iron filings. AB - Field and laboratory experiments were conducted to study the potential use of Fe filings to remove monomethyl arsenate (MMA) and dimethyl arsenate (DMA) from contaminated waters. Batch experiments show that the affinity of MMAfor Fe filings is comparable to that of inorganic arsenate, but lower than that for arsenite. In contrast, very little DMA was retained by Fe filings or their corrosion products. The wide range of affinities of different As species for Fe filings was confirmed by a series of retention and selective-leaching experiments using small columns and As-spiked groundwater. The effectiveness of Fe filings was also demonstrated with a field deployment at a U.S. Superfund site where groundwater is highly contaminated with both organic and inorganic As species. Over the course of 4 months, a 3 L cartridge of Fe filings removed > 85% of As contained in 16000 L of groundwater containing 1-1.5 mg/L total dissolved As, approximately 30% of which was organic As. The results indicate that Fe filings could be used to treat groundwater contaminated with MMA, which is the main organic form of As at most contaminated sites. Indications of partial demethylation in the column of Fe filings suggest conditions might be optimized further to enhance the removal of DMA. PMID- 16245842 TI - Mercury ions removal from aqueous solution using an activated composite membrane. AB - This work presents the results concerning the first use of activated composite membranes (ACMs) for the removal of Hg(II) ions from aqueous solution, using as the ligand di-(2-ethylhexyl)dithiophosphoric acid (DTPA). The effects on the removal percentage of Hg(ll) of variables such as pH, the nature of the acid, the concentration of mercury (in the feed solutions), and the ligand content (in the membrane) as well as the total surface membrane area exposed to the Hg(II) aqueous solution were studied. During the course of the removal experiments, the membrane was immersed in the Hg(II) aqueous solution in acid media and samples of the solution were taken at different times to enable monitoring of the mercury concentration changes. Itwasfound that when the ACM was prepared with a 1.0 M DTPA casting solution and the feed solution contained 2.49 x 10(-4) M Hg(II) in HCI 0.1 M the amount of mercury extracted was higher than 93%. Straight forwardly, additional experiments were carried out with the free-DTPA composite membranes to make up a set of control reference points to verify that removal of the investigated heavy metal was a consequence of the presence of the organic ligand; otherwise there was no Hg(II) concentration variation at all. PMID- 16245843 TI - Secretion of bacterial xenobiotic-degrading enzymes from transgenic plants by an apoplastic expressional system: an applicability for phytoremediation. AB - In search of an effective method for phytoremediation of wastewater contaminated with organic compounds, we investigated the application of an apoplastic expressional system that secretes useful bacterial enzymes from transgenic plants into hydroponic media through the addition of a targeting signal. We constructed transgenic Arabidopsis expressing the aromatic-cleaving extradiol dioxygenase (DbfB), which degrades 2,3-dihydroxybiphenyl (2,3-DHB), and transgenic tobacco expressing haloalkane dehalogenase (DhaA), which catalyzes hydrolytic dechlorination of 1-chlorobutane (1-CB). Although crude leaf extracts of transgenic plants expressing cytoplasm-targeted degradative enzymes showed higher activity than did those from transgenic plants expressing apoplast-targeted enzymes, the hydroponic media of the latter showed 23.2 times (DbfB) and 76.4 times (DhaA) higher activity than plants containing the cytoplasm-targeted enzymes. Addition of crystalline 2,3-DHB to 100 mL of the hydroponic medium of transgenic or wild-type seedlings revealed that only medium from the transgenic Arabidopsis expressing apoplast-targeted DbfB showed rapid ring cleavage of 2,3 DHB. Transgenic tobacco expressing apoplast-targeted DhaA also resulted in the accumulation of the dehalogenation product 1-butanol in the hydroponic medium and showed a higher tolerance to 1-CB than wild-type or transgenic plants expressing cytoplasm-targeted DhaA. These results demonstrate the usefulness of the apoplastic expression of bacterial recombinant proteins in phytoremediation. PMID- 16245844 TI - Advances in seeded ambient temperature ferrite formation for treatment of acid mine drainage. AB - Advances in the seeded ambient temperature ferrite process for treatment of acid mine drainage (AMD) are described. Magnetite formation requires that the oxidation rate of ferrous to ferric does not exceed the rate at which ferrous iron is incorporated into the crystal structure (dehydroxylation crystallization). If the oxidation rate is too high, then ferric-only oxides form, an effect exacerbated by the presence of calcium. NaOH was used to raise the pH of simulated AMD, which also contained calcium so as to simulate the use of lime (i.e., the dissolved Ca/ Fe2+ concentration was maintained at 1:1 bythe coaddition of CaCl2 because this is the Ca/Fe ratio that occurs when pH is elevated by the dissolution of lime). Raising the pH to 10.5 causes Fe2+ to precipitate as "ferrous intermediate" (FI), which is then partially oxidized to magnetite (Fe2+Fe3+2O4). The inhibitory effect of calcium is overcome by a combination of magnetite seed particles, high FI concentrations, and aging. High FI concentrations are easily obtained, even from AMD low in Fe2+, by a contact stabilization reactor-settler sequence. Results for simultaneous removal of cobalt, a metal found in significant concentrations in South African AMD, are also presented. PMID- 16245845 TI - High flux filtration medium based on nanofibrous substrate with hydrophilic nanocomposite coating. AB - A novel high flux filtration medium, consisting of a three-tier composite structure, i.e., a nonporous hydrophilic nanocomposite coating top layer, an electrospun nanofibrous substrate midlayer, and a conventional nonwoven microfibrous support, was demonstrated for oil/water emulsion separations for the first time. The nanofibrous substrate was prepared by electrospinning of poly(vinyl alcohol) (PVA) followed by chemical cross-linking with glutaraldehyde (GA) in acetone. The resulting cross-linked PVA substrates showed excellent water resistance and good mechanical properties. The top coating was based on a nanocomposite layer containing hydrophilic polyether-b-polyamide copolymer or a cross-linked PVA hydrogel incorporated with surface-oxidized multiwalled carbon nanotubes (MWNTs). Scanning electron microscopy (SEM) examinations indicated that the nanocomposite layer was nonporous within the instrumental resolution and MWNTs were well dispersed in the polymer matrix. Oil/ water emulsion tests showed that this unique type of filtration media exhibited a high flux rate (up to 330 L/m2-h at the feed pressure of 100 psi) and an excellent total organic solute rejection rate (99.8%) without appreciable fouling. The increase in the concentration of surface-oxidized MWNT in the coating layer generally improves the flux rate, which can be attributed to the generation of more effective hydrophilic nanochannels for water passage in the composite membranes. PMID- 16245846 TI - Efficient photochemical decomposition of long-chain perfluorocarboxylic acids by means of an aqueous/liquid CO2 biphasic system. AB - Photochemical decomposition of persistent and bioaccumulative long-chain (C9-C11) perfluorocarboxylic acids (PFCAs) with persulfate ion (S2O8(2-)) in an aqueous/liquid CO2 biphasic system was examined to develop a technique to neutralize stationary sources of the long-chain PFCAs. The long-chain PFCAs, namely, perfluorononanoic acid (PFNA), perfluorodecanoic acid (PFDA), and perfluoroundecanoic acid (PFUA), which are used as emulsifying agents and as surface treatment agents in industry, are relatively insoluble in water but are soluble in liquid CO2; therefore, introduction of liquid CO2 to the aqueous photoreaction system reduces the interference of colloidal PFCA particles. When the biphasic system was used to decompose these PFCAs, the extent of reaction was 6.4-51 times as high as that achieved in the absence of CO2. In the biphasic system, PFNA, PFDA, and PFUA (33.5-33.6 micromol) in 25.0 mL of water were 100%, 100%, and 77.1% decomposed, respectively, after 12 h of irradiation with a 200-W xenon-mercury lamp; F- ions were produced as a major product, and short-chain PFCAs, which are less bioaccumulative than the original PFCAs, were minor products. All of the initial S2O8(2-) was transformed to SO42-. The system also efficiently decomposed PFCAs at lower concentrations (e.g., 4.28-16.7 micromol of PFDA in 25.0 mL) and was successfully applied to decompose PFNA in floor wax. PMID- 16245847 TI - Pharmaceutical retention mechanisms by nanofiltration membranes. AB - This study investigates the retention mechanisms of three pharmaceuticals sulfamethoxazole, carbamazepine, and ibuprofen-by nanofiltration (NF) membranes. Laboratory-scale experiments were carried out with two well-characterized NF membranes, with the goal of relating pharmaceutical retention behavior to membrane characteristics, physicochemical properties of the pharmaceutical molecules, and solution chemistry. Results show that retention of pharmaceuticals by a tight NF membrane is dominated by steric (size) exclusion, whereas both electrostatic repulsion and steric exclusion govern the retention of ionizable pharmaceuticals by a loose NF membrane. In the latter case, speciation of pharmaceuticals may lead to a dramatic change in retention as a function of pH, with much greater retention observed for ionized, negatively charged pharmaceuticals. For uncharged pharmaceutical species, intrinsic physicochemical properties of the pharmaceutical molecules can substantially affect their retention. In its neutral form, ibuprofen adsorbs considerably to the membrane because of its relatively high hydrophobicity. Similarly, polarity (represented by the dipole moment) can influence the separation of molecules that are cylindrical in shape because they can be directed to approach the membrane pores head-on due to attractive interaction between the molecule polar centers and fixed charged groups on the membrane surface. This phenomenon is probably inherent for high dipole moment organic compounds, and the governing retention mechanism remains steric in nature. PMID- 16245848 TI - Quantitative structure--property relationships for enhancing predictions of synthetic organic chemical removal from drinking water by granular activated carbon. AB - Granular activated carbon is a frequently explored technology for removing synthetic organic contaminants from drinking water sources. The success of this technology relies on a number of factors based not only on the adsorptive properties of the contaminant but also on properties of the water itself, notably the presence of substances in the water which compete for adsorption sites. Because it is impractical to perform field-scale evaluations for all possible contaminants, the pore surface diffusion model (PSDM) has been developed and used to predict activated carbon column performance using single-solute isotherm data as inputs. Many assumptions are built into this model to account for kinetics of adsorption and competition for adsorption sites. This work further evaluates and expands this model, through the use of quantitative structure-property relationships (QSPRs) to predict the effect of natural organic matter fouling on activated carbon adsorption of specific contaminants. The QSPRs developed are based on a combination of calculated topographical indices and quantum chemical parameters. The QSPRs were evaluated in terms of their statistical predictive ability,the physical significance of the descriptors, and by comparison with field data. The QSPR-enhanced PSDM was judged to give results better than what could previously be obtained. PMID- 16245849 TI - Mechanisms of advanced oxidation processing on bentonite consumption reduction in foundry. AB - Prior full-scale foundry data have shown that when an advanced oxidation (AO) process is employed in a green sand system, the foundry needs 20-35% less makeup bentonite clay than when AO is not employed. We herein sought to explore the mechanism of this enhancement and found that AO water displaced the carbon coating of pyrolyzed carbonaceous condensates that otherwise accumulated on the bentonite surface. This was discerned by surface elemental analysis. This AO treatment restored the clay's capacity to adsorb methylene blue (as a measure of its surface charge) and water vapor (as a reflection of its hydrophilic character). In full-scale foundries, these parameters have been tied to improved green compressive strength and mold performance. When baghouse dust from a full scale foundry received ultrasonic treatment in the lab, 25-30% of the dust classified into the clay-size fraction, whereas only 7% classified this way without ultrasonics. Also, the ultrasonication caused a size reduction of the bentonite due to the delamination of bentonite particles. The average bentonite particle diameter decreased from 4.6 to 3 microm, while the light-scattering surface area increased over 50% after 20 min ultrasonication. This would greatly improve the bonding efficiency of the bentonite according to the classical clay bonding mechanism. As a combined result of these mechanisms, the reduced bentonite consumption in full-scale foundries could be accounted for. PMID- 16245850 TI - Fate of rising CO2 droplets in seawater. AB - The sequestration of fossil fuel CO2 in the deep ocean has been discussed by a number of workers, and direct ocean experiments have been carried out to investigate the fate of rising CO2 droplets in seawater. However, no applicable theoretical models have been developed to calculate the dissolution rate of rising CO2 droplets with or without hydrate shells. Such models are important for the evaluation of the fate of CO2 injected into oceans. Here, I adapt a convective dissolution model to investigate the dynamics and kinetics of a single rising CO2 droplet (or noninteracting CO2 droplets) in seawater. The model has no free parameters; all of the required parameters are independently available from literature. The input parameters include: the initial depth, the initial size of the droplet, the temperature as a function of depth, density of CO2 liquid, the solubility of CO2 liquid or hydrate, the diffusivity of CO2, and viscosity of seawater. The effect of convection in enhancing mass transfer is treated using relations among dimensionless numbers. The calculated dissolution rate for CO2 droplets with a hydrate shell agrees with data in the literature. The theory can be used to explore the fate of CO2 injected into oceans under various temperature and pressure conditions. PMID- 16245851 TI - Change of mechanical properties during short-term natural weathering of MSWI bottom ash. AB - The present work describes the change of mechanical properties during the natural weathering of freshly quenched processed bottom ash. An unconfined uniaxial compression to failure test of the unbound material was used to determine compressive strength and modulus of elasticity. Three main stages of mechanical behavior were determined. In the first stage, during a period lower than 30 days, mechanical properties suddenly increase, with a compressive strength and elastic modulus 7 times greater than the initial parameters. During the second stage, compressive strength and modulus of elasticity lightly increase until approximately 90 days of curing time. Starting from this period both mechanical properties remain steady and independent of the curing time. The neoformed phases, the elevated water content, and the improvement of particle contact after compaction act as a binder layer among particles, increasing the mechanical parameters during the short-term natural weathering process. Because of this, the freshly compacted bottom ash progresses from behaving as an unbound material into a bound pavement material. These mechanical properties obtained from freshly quenched bottom ash are 6-7 times greater than those obtained from previously weathered bottom ash. The bottom ash expansion and leaching of metals were also evaluated. PMID- 16245852 TI - Metal compartmentation and speciation in a soil sentinel: the earthworm, Dendrodrilus rubidus. AB - Earthworms are well-studied organisms in ecotoxicology because of their keystone ecological status and metal-accumulating capacity. However, the direct estimation of the bioreactive fractions of accumulated metal burdens remains technically elusive. In this study we exploited two physical techniques, electron probe X-ray microanalysis (EPXMA) and X-ray absorption spectroscopy (XAS), to improve understanding of the subcellular spatial distributions, ligand affinities, and coordination chemistries of Cd, Pb and Zn in a field population of the epigeic earthworm, Dendrodrilus rubidus. EPXMA and XAS analyses were performed on cryopreparations to maintain compositional fidelity; EPXMA data were analyzed by multivariate statistics. XAS provided whole-worm insights; EPXMA provided in situ, subcellular data from the major metal-sequestering tissue, the chloragog. Both techniques showed that Cd is coordinated with S; the measured Cd-S bond distance in XAS suggests a metallothionein-type ligand. The mean Cd:S molar ratio (EPXMA) of 0.36 is higher than the ratio of 0.29 estimated from published biochemical data. EPXMA and XAS data also found that Ca, Pb, and Zn are predominantly bound to one or more O-donating, probably phosphate-rich, ligands. X-ray distribution maps (EPXMA) of the hepatocyte-resembling chloragocytes revealed that the O-seeking (Ca, Pb, Zn) metals and S-seeking Cd bioaccumulate in distinct organelles. Extended X-ray absorption fine structure showed that the Pb complex is not biogenic pyromorphite, although X-ray absorption near edge structure did not eliminate the possibility. XAS provided no evidence of Pb spillage from the "sequestration compartment" within D. rubidus. However, the correspondence of Pb with Ca and P in EPXMA is not as strong as that of Zn. This is indicative either of spillover or of a second, hitherto unidentified, sequestered-Pb pool. By exploiting the complimentary techniques of EPXMA and XAS,we are closer to describing the mechanistic link between equilibrated body burdens and biomarker responses in earthworms. PMID- 16245853 TI - Confronting workplace exposure to chemicals with LCA: examples of trichloroethylene and perchloroethylene in metal degreasing and dry cleaning. AB - Life-Cycle Assessment (LCA) aims to assess all environmental impacts "from cradle to grave". Nevertheless, existing methods for Life-Cycle Impact Assessment (LCIA) generally do not consider impacts from chemical exposure at the workplace. This is a severe drawback, because neglecting occupational health effects may result in product or process optimizations at the expense of workers' health. We adapt an existing LCIA method to consider occupational health effects from the use of perchloroethylene (PCE) and trichloroethylene (TCE) in dry cleaning and metal degreasing. The results show that, in applications such as metal degreasing and dry cleaning, long-term (steady-state) concentrations at the workplace are up to 6 orders of magnitude higher than ambient air levels. Legal threshold values may be exceeded, depending on machine technology, size, and surrounding working conditions. The impact from workplace exposure to the total human-toxicity potential of the complete life cycle of PCE and TCE (including use, production, and disposal) is accordingly high. We therefore conclude that occupational health effects need to be considered in LCA to prevent overlooking key environmental health impacts in LCA. PMID- 16245854 TI - Risk ranking of bioaccessible metals from fly ash dissolved in simulated lung and gut fluids. AB - Power plant fly ash from two fuels, coal and a mixture of coal and shredded tires, were evaluated for trace metal solubility in simulated human lung and gut fluids (SLF and SGF, respectively) to estimate bioaccessibility. The proportion of bioaccessible to total metal ranged from zero (V) to 80% (Zn) for coal-derived ash in SLF and from 2 (Th) to 100% (Cu) for tire-derived fly ash in SGF. The tire derived ash contained much more Zn. However, Zn ranked only 5th of the various toxic metals in SGF compared with international regulations for ingestion. On the basis of total concentrations, the metals closestto exceeding limits based on international regulations for inhalation were Cr, Pb, and Al. On dissolution in SLF, the most limiting metals were Pb, Cu, and Zn. For metals exposed to SGF there was no relative change in the top metal, Al, before and after dissolution but the second-ranked metal shifted from Pb to Ni. In most cases only a proportion of the total metal concentrations in either fly ash was soluble, and hence bioaccessible, in either biofluid. When considering the regulatory limits for inhalation of particulates, none of the metal concentrations measured were as hazardous as the fly ash particulates themselves. However, on the basis of the international ingestion regulations for Al, the maximum mass of fly ash that could be ingested is only 1 mg per day (10 mg based on bioaccessibility). It is possible that such a small mass could be consumed by exposed individuals or groups. PMID- 16245855 TI - Comment on "Gonadal development of larval male xenopus laevis exposed to atrazine in outdoor microcosms". PMID- 16245856 TI - [Phylogenetic analysis of the surface glycoprotein genes of an aquatic bird origin influenza virus isolate A/Duck/Yangzhou/233/2002 (H6N2)]. AB - Several H6 subtype avian A influenza viruses were isolated from aquatic birds in some live bird markets when we surveyed the ecology of the influenza in East China for more than two years and identified by specific RT-PCR. In this paper, the hemagglutinin (HA) and neuraminidase (NA) gene of one representative virus named A/Duck/Yangzhou/233/ 2002 (H6N2) (Dk/YZ/233/02) had been sequenced. Phylogenetic analysis of the H6 gene sequences available in the Influenza Sequence Database showed that the Dk/YZ/233/02 viruse cluster together with Dk/HK/3461/99 (H6N1) and Ck/Taiwan/na3/98 (H6N1). Phylogenetic analysis of the N2 NA genes of Dk/YZ/233/02 revealed that the NA gene of Dk/YZ/233/02 had genetically close relationships with that of H9N2 viruses isolated from duck in Japan and from chickens in South Korea, which were distinct from those of Ck/Beijing/94 (H9N2). The sequence of cleavage site between HA1 and HA2 of Dk/YZ/233/02 is P-Q-I-E-T-R-D, which was the typical characterization of the LPAIV. PMID- 16245857 TI - [Cloning and characterization of an harpin-encoding gene from Xanthomonas axonopodis pv. glycines required for hypersensitive response on nonhost plant tobacco]. AB - An hpa1 gene was cloned into an expression vector, pET30a(+), from the genomic DNA of Xanthomonas axonopodis pv. glycines (Xag), the causal agent of soybean bacterial pustule, with degenerated primers by polymerase amplification reaction (PCR). The gene product was extracted from the conjugate (BHR-3) of BL21 (DES) with the recombined vector pHR3 after the engineering strain was induced by IPTG in LB medium. The SDS-PAGE gel showed that the gene product was 15.1kD. The product was heat-stable (10 min at 100 degrees C), protease K sensitive, and able to trigger hypersensitive response (HR) in common tobacco, but was unable to elicit HR in NahG transgenic tobacco in which salicylic acid accumulation was abolished. Moreover, the HR elicitation of the protein in tobacco was dispelled by eukayotic metabolic inhibitors, actinomycin D, cycloheximide and LaCl3. The 402 bp hpa1 gene in this study putatively encoded a 133 ammonia acid protein of which glycine (G) was rich with 21.1%. Sequence comparison indicated that the hpa1 gene and its protein was 51.4% - 93.8% identity with those of Xanthomonas oryzae pv. oryzae and other Xanthomonas species and pathovars. Alignments of harpin proteins of Xanthomonas genus displayed that the glycine-rich region with GGG-GG motif was variable. The comparison also showed that the harpin-encoding gene of Xag (nominated here as hpa1(Xag)) did not possess any similarity with that of Erwinia amylovora, Pseudomonas syringae and Ralstonia solanacearum at nucleotide and protein levels. It is concluded that hpa1(Xag) gene encodes an harpin protein which elicits a typical HR in nonhost tobacco. PMID- 16245858 TI - [Effect of lat disruption on clavulanic acid production]. AB - A 1.8kb fragment of lat was obtained from Streptomyces clavuligerus 27064, and replacement plasmid of pXAL1 and pXAL2 were constructed. PXAL1 and pXAL2 were used to disrupt the lat gene by bi-parental conjugation from E. coli to Streptomyces clavuligerus. A Am(r)Thio(S) transformant, named as XAL863, was obtained. The genome of Streptomyces clavuligerus 27064 and XAL863 was analyzed by southern blot technique, and the activity of lysine epsilon-aminotransferase in the two strains was also tested. Both results proved that the lat was disrupted in the XAL863. Streptomyces clavuligerus and XAL863 were cultured in the shaken flask respectively, and the production of clavulanic acid was analyzed by HPLC with the different incubation time interval, and the yield was approximately 1.8 times higher in the XAL863 at their highest production point. PMID- 16245859 TI - [Cloning and expression of P450 genes CYPA14 and CYPA16 from Candida tropicalis 1230]. AB - Cytochrome P450 (CYP) as a monooxygenase catalyzes the limiting-rate reaction in the oxidation pathway of Candida tropicalis (C. tropicalis). Two members of P450 gene CYPA16 and CYPA14 were isolated from C. tropicalis 1230 by cassette PCR method, based on cloning of the P450 partial gene. They are tandemly arranged on the chromosome, encoding the gene products of 522 and 540 amino acid residues respectively. The deduced amino acid sequences of CYPA14 and CYPA16 are identical to those of CYP52A14 and CYP52A16 respectively,and highly similar to those of CYP52A2 and CYP52A1 from C. tropicalis ATCC 750 respectively. Simultaneously, CYPA14 and CYPA16 from several dicarboxylic-acid-producing strains were also cloned, and some site-mutations were found in their sequence. CYPA14 and CYPA16 were expressed in the Saccharomyces cerevisiae strain. It was shown that the recombinant P450 content of CYPA14 was lower than that of CYPA16, and part of them were denatured. Comparing to C. tropicalis 1230, no significant changes were observed in recombinant P450 contents of CYPA14 and CYPA16 from every dicarboxylic-acid-producing strain. PMID- 16245860 TI - [Influence of expression of transglutaminase on the growth of Lactococcus lactis]. AB - To improve the aerobic growth performance of Lactococcus lactis subsp. cremoris NZ9000, the gene mtg encoding the mature microbial transglutaminase was amplified from the chromosomal DNA of Streptoverticillium mobaraense and then cloned into the nisin-inducible expression vector pNZ8148. The resulting plasmid pFL001 was transformed into strain NZ9000 by electroporation. Compared with strain NZ9000 harboring pNZ8148 (the control strain), strain NZ9000 harboring pFL001 (the recombinant strain) had a remarkably improved aerobic growth performance. When grown aerobically under non-pH-controlled conditions, the maximal biomass of the recombinant strain reached 4.13 g/L, which was 11-fold higher than the growth of the control strain (0.34 g/L). When grown aerobically with the pH controlled at 6.5 +/- 0.1, the maximal biomass of the recombinant strain reached 4.73 g/L, which was an 80% increase compared with the growth of the control strain. In addition, the efficiency of biomass synthesis relative to glucose consumption (Y(x/s)) of the recombinant strain, 71.7 g of biomass per mol of glucose, was 1.6 fold higher than that of the control strain. The significantly improved growth performance of the recombinant strain might be attributed to the expression of mtg in the recombinant strain, which might increase intracellular pH and save part of the energy(ATP) that was originally used for pumping the cytoplasmic H+, and as a consequence, the energy used for growth increased accordingly. PMID- 16245861 TI - [Antibacterial spectrum of antibacterial peptides from Musca domestica larvae and synergic interaction between the peptides and antibiotics]. AB - In vitro antibacterial activities of three antibacterial peptides, alone and in combination with the antibiotics were measured by MIC and antibacterial spectrum. Synergy was observed when the peptides were combined with penicillin, streptomycin by FIC. The results show that antibacterial peptides have stronger inhibition of pathogenic bacterium, and antibacterial spectrum are very different. The interaction in antibacterial peptides, antibiotics and bacterium is extraordinary complex, except synergic action, there are antagonism and indifference. PMID- 16245862 TI - [Effect of high hydrostatic pressure on microbial physiological characteristics]. AB - Physiological characterizations of Listeria monocytogenes NCTC 11994 and Escherichia coli ATCC 80739 have deeply changed by high hydrostatic pressure. The results showed that counts of both microbial strains decreased 7 log cfu at 400MPa, 10 min. Pressure treatments also resulted in change of Intracellular pH value, lowed membrane potential, have internal potassium filtered out, and decreased ATP concentration. PMID- 16245863 TI - [Mechanism study on difference of biotransformation between Mycobacterium fortuitum MF2 and MF96]. AB - Biotransformation difference between parent strain (MF2) and mutant strain (MF96) of Mycobacterium fortuitum was observed. Biotransformation with resting cells showed that the major products of biotransformation by both parent and mutant strains are delta4-androstenedione(4AD) and testosterone(TS). Experiments with cell-free extract system showed that the proportion of 4AD/TS obtained from parent and mutant strains was almost same when enough NAD+ and NADH were supplied in this system. It was suggested that the difference of the ratio of products transformed by both strains in resting cell system may result from their different ratio of NAD+/NADH. This speculation was verified to be true by determination of the amount of NAD+ and NADH presented in both strains. PMID- 16245864 TI - [Expression of feedback-resistant aspartate kinase gene in Corynebacterium crenatum]. AB - The AEC-resistant aspartate kinase gene from C. crenatum CD945 was cloned into vector pJC1. Its expression was investigated both in the wild type C. crenatum AS1.542 and its mutant C. crenatum CD945. The result showed that C. crenatum AS1.542 harboring AK(fbr) gene could grow on the defined medium with the co existence of 12 mg/mL both of AEC and L-threonine respectively. Overexpression of AK(fbr) gene in C. crenatum CD945 results in a 4-fold increase of specific enzyme activity than the parental strain. The amplification of the activity of aspartate kinase yields 22% increase of L-lysine production and 23% increase of L-lysine productivity without affecting the growth rate. PMID- 16245865 TI - [Study on breeding of probiotic Aspergillus nigre by N+ implantation and fermentation condition of its mutant]. AB - Using Aspergillus niger AN01 as original strain, mutated strain Aspergillus niger AN03 was obtained by N+ implantation. The results showed that activities of acidic protease, cellulase and pectinase of Aspergillus niger were raised from 71.6U/g, 141.7U/g and 264.8U/g to 996.5U/g, 940.4U/g and 906.5U/g respectively. Characteristics of enzymes production in the mutant Aspergillus niger AN03 kept stabile by 5 times subculture. In addition, the optimum conditions for enzymes production of Aspergillus niger AN03 were investigated. The optimum components of medium consisted of bran 105.0g, corn straw 105.0g, bean cake 105.0g, NH4Cl 6.0g, H2O 1000mL. The optimum fermentation condition was incubated for 4d in the condition of pH 5.0 and temperature 30 degrees C. PMID- 16245866 TI - [Cloning and expression of an Aspergillus fumigatus chitosanase gene]. AB - According to published DNA sequence of Aspergillus fumigatus chitosanase(Csn) gene, 8 long single DNA strands each about 100bp and 4 DNA primers were designed and synthesised. By PCR, 8 DNA strands were connected into a complete chitosanase gene of 624bp. This chitosanase gene was not identical with its wild type, some point mutations were introduced into its DNA sequence by special design of those 8 DNA strands. These mutaions did not change amino acid composition of the chitosanase, however, the codens were changed into E. coli favorites. The Csn gene was cloned into plasmid pGEM-Teasy and verified by DNA sequence analysis. Thereafter, Csn gene was subcloned into a fusion-protein expressing vector pGEX 3X. Recombinant plasmid pGEX-Csn was transformed into E. coli DH5alpha and the transformant was induced expressing with 0.5 mmol/L IPTG. Expressing product was analyzed by SDS-PAGE, fusion protein GST-Csn was purified by affinity chromatography. By factor X a digestion GST-Csn was cleaved and GST was taken out by another chromatography. The biological activity of recombinant chitosanase(rCsn) was also detected, as a result the recombinant Csn had a strong ability of degrading chitosan, which was much higher than lysozyme. Its chitosan degradation activity could be influenced by pH and temperature. PMID- 16245867 TI - [Cloning and expression in Pichia pastoris of an alkaline mannanase gene]. AB - A strain containing alkaline mannanase gene was isolated from soil by functional plates and the genome library was constructed. From it a mannanase gene TM1 was acquired and was sequenced. The BLAST analysis showed a lower-than-60% similarity of the amino acid sequence to those in GenBank and proved TM1 to be a new mannanase gene (GenBank accession number AY623903). The new gene without signal peptide was cloned into the Pichia pastoris expression vector pHBM905C. The recombinant plasmid pHBM1201 was digested by Sal I and transformed into Pichia pastoris KM71, GS115, SMD1168, respectively. All of the recombinant Pichia pastroris strains containing pHBM1201 secreted functional beta-mannanase. Because of its high mass of expression, the recombinant Pichia pastoris SMD1168-3 containing pHBM1201 was induced at shake flasks. The optimal temperature and pH of the beta-mannanase produced by the recombinant strains were 55 degrees C and 7.5, respectively. The enzymatic activity for konjak powder reached 41.8 with a half life of one hour. After keeping at 80 degrees C for 5 min, the enzymatic activity declined from 77% to 11% and the enzymatic activity could recover up to more than 60% when the temperature descended to 55 degrees C. PMID- 16245868 TI - [Purification and properties of recombinant extremely thermostable and acid stable amylase]. AB - Extremely thermostable and acid-stable a-amylase produced by Pichia pastoris GS115/pPIC9K-Amy-228 was purified to electrophoretic homogeneity by the steps of ultrafiltration and PAGE. Purification of about 11.7 fold was achieved with an overall yield of 29.8%. Its molecular weight was estimated to be about 55kD by SDS-PAGE. The isoelectric point was 5.0 (room temperature). Michaelis constant of the enzyme for soluble starch was 1.12g/L. The carbohydrate content was 15.4% by the phenol-sulfuric acid method. The optimum temperature and pH of the enzyme activity were 95 degrees C and 4.5 respectively. The enzyme activity was stable under room temperature in the pH rang of 4.0 - 7.0 for 48 hours. About 60% of the initial enzyme activity was measured after 1h of incubation at 110 degrees C. The activity was strongly inhibited by Fe2+, Cr2+ and Cu2+, While Ca2+ had no effect on it. DTT and EDTA had no effect on the activity. PMID- 16245869 TI - [Induction of nicotinic acid hydroxylase activity of Pseudomonas putida NA-1 and optimization of transformation conditions]. AB - Some features of Pseudomonas putida NA-1 strain in cultivation and enzyme production were different from S. marcescens IFO 12648 and P. fluorescens TN5 stains which could transform nicotinic acid to 6-hydroxynicotinic acid reported by other scientists, such as optimal carbon and its optimal concentration, optimal inducer concentration, and optimal cultivation temperature. The ideal transformation condition was nicotinic acid 3%, temperature 35 degrees C and pH 7.0. Under an appropriate condition, in a 4 liter fermenter, production yield of 6-hydroxynicotinic acid by Pseudomonas putida NA-1 was 108.39 g/L. PMID- 16245870 TI - [Cloning and analysis of genes encoding 2-naphthoate monooxygenase and NADH:flavin oxidoreductase]. AB - In Burkholderia sp. JT1500, a key step of 2-naphthoate biodegradation pathway is carried out by 2-naphthoate monooxygenase (Nmo) in which 2-naphthoate is oxidized to 1-hydroxy-2-naphthoate. A gene cluster of 4.8kb from Burkholderia sp. JT1500 was cloned and sequenced, four open reading frames named orfB, orfC, orfD and orfA were identified in this region. Sequence alignment showed that orfA had a high homology of nucleotide acid composition to monooxygenase genes from both Japonicum USDA 110 and Ralstonia eutropha HF 39, orfB had some homology to the component of flavin reductase genes from Bordetlla pertussis Tohama I, Ralstonia solanacearum GMI1000 and Bordetella bronchiseptica RB50. Enzyme activity analysis showed that the cell extracts of recombinant E. coli S(A) (only harboring orfA) showed very low oxygenase oxidation activity as detected by NADH decreasing, while the cell extracts of recombinant S(B) (only harboring orfB) did not show any oxidation activity at all. But when the cell extracts of S(B) and S(A) were mixed, which showed very strong oxidation activity when flavin (FMN or FAD) provided; the recombinant S(B + A) cells harboring both orfB and orfA genes also showed strong oxidation activity when flavin provided; weak flavin deoxidization activity could be detected from the cell extracts of E. coli S(B) under anaerobic conditions. Based on above message, a conclusion was drawn that Nmo is consisted of two components: a flavin oxidoreductase (NmoB) and a monooxygenase (NmoA). First NmoB uses NADH to reduce flavin and supplies reduced flavin to NmoA to catalyze O2 oxidizing 2-NAT. NmoB is NmoA' s coupling protein. PMID- 16245871 TI - [Analysis of signal peptides of the secreted proteins in Agrobacterium tumefaciens C58]. AB - The 4554 ORFs of Agrobacterium tumefaciens C58 Cereon were used for the prediction of signal peptides by the network tools, such as SignalP3.0, LipoP1.0, TMHMM2.0 and TargetP1.01. Total 203 signal peptides with conserved amino residues are found, among them, 158 are secretary types, 9 are RR-motif types, 28 are SignalPase II types and 8 are bacteriocin-pheromone types. However, only two signal peptides from the secreted proteins, AGR-C-1878p and AGR-C-1880p have the same amino sequences, showing the signal peptides of the strain are highly variable. PMID- 16245872 TI - [Analysis of the probiotic Bifidobacterium and Lactobacillus community in child intestinal flora]. AB - To investigate the distribution of child intestinal flora and the composition of its key probiotics community, study on intestinal flora of 21 Chinese children (age 2 - 5) was conducted, which included bacteria isolation and counting, 16S rDNA sequencing and homology analysis. For identification of the key probiotics such as Bifidobacterium and Lactobacillus in children feces at the species level, the specific primers Im26/Im3 and L159/L677 for PCR amplification of partial 16S rDNA were used. The results show that the composition of child intestinal flora is was relatively stable and almost same to the intestinal flora of the youth (age 20 - 25). Culture-based approaches show that the key probiotic community in feces at the species level was highly different in composition and numbers from individual to individual. B. longum and B. pseudocatenulatum, which are detected at levels of 10(7) CFU/g (wet) in samples and the detection rates are 90.48% and 85.71% respectively, are believed to be major bifidobacterial species in child intestinal microbiota. In addition, B. adolescentis, B. bifidum, B. infantis and B. thermacidophium have also been found. L. mucosae, L. fermentum, L. salivarius, L. ruminis, L. gasseri and L. plantarum are isolated from the stools. L. mucosae (3.68 log10 CFU/g (wet), detection rate 71.43%) and L. fermentum (3.97 log10 CFU/g (wet), detection rate 52.38%) are two dominant species of Lactobacillus. Study on Chinese child intestinal flora, especially on the compositions and numbers of key probiotics in the feces will be very helpful to the development of effective probiotics in future. PMID- 16245873 TI - [Study of bacteria-lysis abilities and growth conditions of 4 Bdellovibrio]. AB - Bdellovibrio can lyse pathogenic bacteria and clean up waters. 4 strains of Bdellovibrio sp., designated Bh04-4, Bh04-41a, Bh04-A + and Bh04-1f, were isolated from seawaters used Bh04 as host bacterium. After confirmation to be Bdellovibrio sp. by electron microscopy and specific PCR method, their growth conditions and lytic ability on 61 bacteria from various sources were performed. Results showed that all four Bdellovibrio grew in salinity in the range of between 1% and 3%, with 3% salinity being the most suitable one. They grew in the range of temperature from 15 to 30 degrees C, with 20 - 25 degrees C as their best growth temperature. These four Bdellovibrio only grew on live host bacteria rather than the dead ones. When 61 strains of bacteria were used as hosts, Bh04-4 lysed 21 strains, corresponding to 34.4% of lysis ability (21/61), Bh04-41a lysed 24 strains (39.3% lysis ability), Bh04-A + lysed 40 strains (65.6% lysis ability) and Bh04-1f 43 strains (70.5% lysis ability). Taken all four Bdellovibrio together, they lysed 55 out of 61 strains, amounting to 90.2% lysis ability. Results fully demonstrate the potential application of Bdellovibrio in lysing pathogenic bacteria from the marine environments. PMID- 16245874 TI - [The innate ability of Rhodococcus sp. SDUZAWQ to tolerate sulfur in petroleum]. AB - Removal of sulfur from petroleum can be accomplished by various means. One method is to use microorganisms, such as bacteria. In the present study, strain Rhodococcus sp. SDUZAWQ was employed to test the effects of various concentrations of dibenzothiophene (DBT) and sulfate, had on this removal process. Desulfurization was accomplished, using Basal Salts Medium (BSM), supplemented with 0.2mmol/L DBT and different concentrations of Na2SO4. Growth of SDUZAWQ was pronounced, even when the concentration of DBT was increased to 6mmol/L. Furthermore, it should be noted that the end product of DBT desulfurization, 2 hydroxybiphenyl (2-HBP), was detected as well. This finding was significant because it demonstrated the bacteria' s ability to withstand high concentrations of organosulfur. Dibenzothiophene was utilized when both DBT and Na2SO4 were present in the culture medium. Additionally, 2-HBP was produced. These data are in contrast to previous studies that indicated that DBT could not be metabolized by Rhodococcus sp. in the presence of sulfate. Finally, cloning and sequencing of the gene cluster dszABC, its upstream regulatory sequence and dszD, demonstrated that they share 99%, 100% and 100% with those of R. erythropolis IGTS8, respectively. PMID- 16245875 TI - [Production of monoclonal antibodies to Lily symptomless virus and application in lily virus detection]. AB - Four hybridoma cell lines, 2A2, 5H9, 5H2 and 5E12, secreting monoclonal antibodies (MAbs) against Lily symptomless virus (LSV) were produced by fusing mouse myeloma cells (SP2/0) with spleen cells from BALB/C immunized by the LSV particles. The four MAbs could specifically react with LSV. The titres of ascitic fluids of the four MAbs are up to 10(-6) in ELISA. Isotypes and subclasses of 5H9 and 5E12 belong to IgG1 while those of 2A2 and 5H2 belong to IgG3. Isotypes of light strains of the four MAbs all belong to kappa. The four MAbs were used in antigen-coated plate (ACP)-ELISA for LSV detection, and ACP-ELISA could successfully detect 1.8 ng of purified LSV or virus in plant sap diluted 1:300. The presence of LSV in field lily tissues was investigated with ACP-ELISA. PMID- 16245876 TI - [Emerging of avian leukosis virus subgroup J in a flock of Chinese local breed]. AB - Myeloid leukosis (ML) cases were first diagnosed in a chicken flock of Chinese local breed in Shan dong province. The main symptom included wasting, weight loss, anemia. It caused about 10% mortality of about 15000 birds at the age of 120-day. In the necropsy, gray-white nodules and protrusions in various sizes were commonly observed on the surface of the sternum, intestine and trachea. Almost all viscera tissues showed moderate to severe enlargement with diffuse gray-white nodules. Histological examination indicated that the tumor cells proliferated in tissues were myelocytes with eosinophilic granules in cytoplasm. In PCR with a pair of ALV-J-specific primers, 15 of 17 liver samples were positive. PCR product of one positive sample was sequenced and demonstrated 98.05% and 97.4% identity with ALV-J HPRS-103 strain at nuclei acid and amino acids level, respectively. By immunohistochemistry (IHC) technique with ALV-J monoclonal antibody, the most intense staining was in the tumor tissue, liver, spleen, kidney, bone marrow, and proventriculus. The results indicate that ALV-J already caused chickens infection and dead in Chinese local breed. PMID- 16245877 TI - [Construction of recombinant canine adenovirus type 2 expressing Canine coronavirus spike glycoprotein and its immunogenicity]. AB - In order to construct a recombinant Canine adenovirus type 2 (CAV-2) expressing the spike glycoprotein of Canine coronavirus (CCV), the S1 gene fragment of CCV strain DXMV, encoding major antigenic region A, B, C and D of S protein, was amplified by RT-PCR and cloned into pVAX1 vector. The complete S1 expression cassette was subcloned into the shuttle vector pVAXE3, then further cloned into the backbone vector pPoly2-CAV2 containing complete genome of CAV-2. To gain the recombinant Canine adenovirus, the recombinant plasmid pCAV-2-CCV-S1 was linearized by Cla I/Asc I to release recombinant genome, and then transfected into MDCK cell. The recombinant virus CAV-2-S1 was gained through 4 passages in MDCK, which showed classical CPE of CAV-2. The expressed S1 protein of CCV, which was identified by RT-PCR and Western blot, can be specifically recognized by polyclonal antibody against CCV. The immunization in dogs indicated that the recombinant CAV-2 could effectively induce the specific antibodies against CCV and CAV. PMID- 16245879 TI - [Study on dengue virus infection of human dendritic cells]. AB - To study Dengue virus (DV) infection of human dendritic cells (DC). Monocyte isolated from healthy human peripheral blood were incubated in medium with GM-CSF and IL-4 for more than 7 days. DCs were then collected and identified by transmission electron microscope, immunohistochemistry and lymphocytes stimulatory ability. Dengue virus type II (DV-2) were infected with human dendritic cells(DC) in vitro, culture supernatants and cells were collected by different time postinfection (6h, 12h, 24h, 48h, 72h). Viral titers were evaluated by microplaque forming assay on C6/36 monolayer cells; DV antigen in human dendritic cells were demonstrated by an indirect immunofluorescent assay (IFA). Localization of DV in DC was observed under a transmission electron microscope. The viruses were detected in the culture supernatants as early as 6h after infection; the highest viral titers were obtained at 48h, and then declined to very low titers at 96h. DV-2 antigen was detected in infected DC by IFA. After infection for 48h, DV particles were obvious in cystic vesicle, vacuoles . Human dendritic cells are targets of dengue virus infection. DV could efficiently infect DC and produce virus particles, DC possibly plays a role in the pathogenesis of DV infection. PMID- 16245878 TI - [Immunogenicity of envelope glycoprotein gene of reticuloendotheliosis virus expressed in insect cell]. AB - A recombinant baculovirus expressing reticuloendotheliosis virus env gene was constructed with Bac-to-Bac Baculovirus Expression Systems. After transfecting the recombinant virus into Sf9 cells for 3 days, REV env can be detected by indirect immunofluorescence antibody assay (IFA) and Western blot with specific monoclonal antibodies of REV. The oil-water emulsion vaccine was then produced using this infected Sf9 cells lycates and inoculated SPF chickens to validate the immunogenicity for REV. The results show that special anti-REV antibody can maintain more than 45 days and resist the infection of REV viruses. This is the first success to induce anti-REV antibody in chickens by none-live viruses. PMID- 16245880 TI - [Analysis of secreted proteases of Trichophyton rubrum]. AB - Secreted proteases are thought as potential virulent factors of Tricophyton rubrum. Based on cDNA libraries of 6 physiological phases of Tricophyton rubrum, 9683 unique ESTs were obtained by DNA sequencing. By bioinformatic analysis, 18 ESTs of putative secreted proteases were obtained from the established ESTs library, including 4 secreted peptidases, 1 secreted metalloprotease, 2 extracellular serine proteases, 1 secreted aspartic proteinase, 9 secreted subtilisin-like proteases and 1 vacuolar serine protease. These secreted proteases are related to nutrient uptake, lesion extension and host immune responses elicitation in the process of T. rubrum infection. These results provide a clue to further research on the infection and pathogencity of T. rubrum. PMID- 16245881 TI - [16S rDNA RFLP analysis of endophytic bacteria from Brassica napus]. AB - Endophytic bacteria reside in plant hosts without causing disease symptom. In this study, 122 endophytic bacteria strains isolated from healthy oilseed rape plant were categorized preliminarily to 35 groups based on colony morphology; 16S rDNAs were amplified with universal primers of bacteria, all the strains could produce a single band about 1500bp, the PCR products were digested with restriction endoenzyme Hae III and Msp I respectively, different patterns were obtained for each enzyme, by combining all the restriction patterns for the two enzymes, the 122 strains could be grouped into 39 16S rDNA genotypes. The results indicated that endophytic bacteria from Brassica napus were genetically diversity. By comparing two methods, colony morphology only showed limited and preliminary information, and PCR-RFLP could be as a rapid and accurate tool for the diversity analysis of endophytic bacteria from Brassica napus. PMID- 16245882 TI - [Studies on construction of artificial mutants of Cucumber mosaic virus satellite RNA and their biological activity]. AB - Based on the full length cDNA clone of a Cucumber mosaic virus satellite RNA, which was 369nt in size, artificial mutants were developed by the method of error prone PCR and DNA shuffling. The new satellite cDNAs were transcribed in vitro into ssRNA and pseudo-recombined with a helper Cucumber mosaic virus, which contains no satellite RNA. Sequence analysis showed that A to T/G or G to A replacement all the four mutants, named MS1, MS5, MS6 and MS11 respectively, and there is no C to G or G to C replacement, but amongst, only the mutants MS11 could replicated when recombined with the helper virus strain. No satellite RNA could be detected by RT-PCR amplification and double-stranded RNA analysis for those pseudo-recombination constitution of Cucumber mosaic virus strain with mutants MS1, MS5 and MS6.Sequence homological comparison showed that the single replacement of mutants MS1, MS5 and MS6 occurred in the highly conservative regions and the T to A replacement of mutant MS11 was located in the normal variation region. This is the first artificial mutation of satellite RNA of plant RNA viruses. The results indicated that single base in the region of satellite RNA maybe important to maintaining the biological activity of satellite RNA for its replication and stability. The variation and evolution of satellite RNA could be hopefully studied through combination directed evolution by DNA shuffling with pseudo-recombination in vitro. PMID- 16245883 TI - [PCR site-directed mutagenesis of avian influenza virus hemagglutinin gene in vitro and expression in 293T cell]. AB - The site-directed mutagenesis of HA gene was made by using PCR, and mismatches were introduced into primers. Mutagenesis was performed in a three-step PCR. The amplified fragments from the second PCR which contained the mutation site were cloned into the pcDNA3 vector, named pHAm. The sequencing analysis showed that the mutation site was correct. The amino acid sequence at the cleavage site of the HA protein was from RKKR decrease GLF to RSSR decrease GLF. The recombinant plasmid pHAm was transiently transfected into 293T cells by the calcium phosphate precipitation method. Indirect immunofluorescent assay (IFA), confirmed expression of the HA protein on the cell membrane, the mutant HA gene was a promising candidate for further studies. PMID- 16245884 TI - [Torulopsis glabrata neomycin-resistant mutant abolishes pyruvate production with enhancement of glucose consumption rate]. AB - To further increase the rate of glucose consumption by multi-vitamin auxotrophic yeast Torulopsis glabrata. A neomycin-resistant mutant N07, with the activity of F1-ATPase decreased roughly 35% but glucose consumed per cell was increased 38% than that of parent strain, was breed based on analysis of energy metabolic pathway. The typical inhibitors of F1F0-ATPase, DCCD, NaN3 and neomycin, depressed the F1-ATPase activity of parental strain but no effect on that of mutant strain. Strain N07 was cultured in a pyruvate fermentation medium containing 100g/L of glucose using flask. It was found that the rate of glucose consumption and pyruvate production were higher by 34% and 42.9% in the mutant than in the parent, respectively. However, the rate and yield of growth (about 24%) of the mutant was lower than that of the parent. The content of intracellular ATP of the mutant also decreased 23.7% than that of the parent. The activities of key enzymes in glycolytic pathway and electron transfer chain of the mutant and the parent were determined. Enzymatic analysis revealed that, compared with the parent strain CCTCC M202019. The activities of key enzymes, phosphofructokinase, pyruvate kinase, glyceraldyde-3-phosphate dehydrogenase of the mutant N07 increased 63.7%, 28.8% and 14.4%, respectively, all the key enzymes of electron transfer chain in the mutant N07 also increased roughly 10%. PMID- 16245885 TI - [The distribution and viability changes of the Agrobacteria in the vacuum infiltrated plants of Chinese cabbage]. AB - To understand further the mechanism of agrobacteria transformation by vacuum infiltration, the distribution and viability changes of the agrobacteria in infiltrated Chinese cabbage( B. rapa ssp. chinesis) plants were investigated by histochemical staining and bacteria plating techniques. Results revealed that there were different quantities of Agrobacteria in different organs, among which the most quantity occurred in flowers, especially in ovules. The Agrobacteria lied in the space between cells and concentrated in the vascular bundle no matter the type of the organs examined. The CFUs (colony forming unit) of the Agrobacteria derived from the flowers were always the most in every experiment. The viabilities of the agrobacteria in the plants declined dramatically along with the elongation of the time after plant infiltration in all organs, but there were still 10(3) CFUs per gram fresh tissue in the flowers after 15 days of infiltration. These results may explain the fact that the ovules were the targets for the Agrobacteria mediated vacuum infiltration transformation. PMID- 16245886 TI - [Expression of a laccase gene from Pleurotus ostreatus in Pichia pastoris and characterization of the recombinant enzyme]. AB - A Pleuotus ostreatus laccase gene was cloned by RT-PCR and designated as lccPol. Its sequence was submitted to GenBank with the accession number AY450404 obtained. The open reading frame was transformed into three Pichia pastoris strains GS115, KM71 and SMD1168, respectively, under control of the AOX1 promoter by using the vector pHBM906. LCCPo1 can be expressed by all three P. pastoris recombinant strains. Three different strategies for shake-flask cultures were compared: (1) (25 degrees C, 1.0% methanol), (2) (20 degrees C, 1.0% methanol), (3) (20 degrees C, 0.5% methanol). The laccase activity could be improved by increasing the methanol concentration befittingly. The results showed that the cultivation temperature had a marked effect on the production of active heterologous laccase. 2 - 6 folds higher laccase activities were obtained when the cultivation temperature was kept at 20 degrees C instead of 25 degrees C. The highest activities, 3.19U/mL [GS115 (pHBM565)], 2.56U/mL [KM71 (pHBM565)], and 2.49U/mL [SMD1168 (pHBM565)], were gotten when the induction were performed at 20 degrees C with 1.0% (V/V) methanol supplied. The temperature and pH optimum for the recombinant laccase produced by three strains were 60 degrees C and pH4.2, respectively. PMID- 16245888 TI - [Studies on extracellular virulent factors produced by potential pathogenic bacteria isolated from abalone postlarvae and biofilms]. AB - To find out the potential pathogen(s) that caused massive death of abalone postlarvae( Haliotis diversicolor supertesta ) in Southern China, 105 bacterial strains were isolated from the water, whitened postlarvae and their biofilms of an abalone farm in Guangdong Province. Extra-cellular protease, gelatinase, lipase as well as haemolysis tests were performed on them. Lysophospholipase (Tlh)-targeted PCR was also carried out in order to reveal if the haemolysis caused by bacterial strains were related to Tlh. Results showed that 35 out of 105 strains possessed extra-cellular enzymatic abilities and strains 1, 2, 3, 5, 9 and 16 were the most powerful ones of all. Among these 35 strains, 85.6% strains (30/35) possessed haemolytic activities on goat blood agar and specific PCR revealed that 16 strains were PCR positive, indicating that they contained Tlh genes. In order to identify these strains, API 20 E strips were employed and results indicated that 50% were vibrios, among them Vibrio alginolyticus composed 70% of the vibrio population. In conclusion, compared to other strains, 6 Vibrio alginolyticus (strain 1, 2, 3, 5, 13, and 16 ) and 2 Vibrio parahaemolyticus (strain 9 and 21) which all were isolated from whitened abalone postlarvae, warrant further studies due to their better abilities in extra-cellular enzyme(s) production and/or haemolytic activities. PMID- 16245887 TI - [Immobilization of laccase on magnetic chitosan microspheres and study on its enzymic properties]. AB - Laccase was immobilized on magnetic chitosan microspheres by using glutaraldehyde as cross-linking reagent. The immobilization conditions and characterization of the immobilized enzyme were investigated. immobilization conditions for laccase were: 10mL of 0.8mg/mL of laccase in phosphate buffer(0.1mol/L, pH 7.0) reacted with 50mg of magnetic chitosan microspheres at 25 degrees C for 1h and subsequently was kept at 4 degrees C for 2h. The immobilized enzyme exhibited the maximal activity at pH 3.0. The optimal temperatures for immobilized enzyme were 10 degrees C and 55 degrees C. The Km value of immobilized laccase for ABTS was 171.1 micromol/L in pH 3.0 phosphate buffer at 37 degrees C. Compared with free enzyme, the thermal, operational, and storage stabilities of the enzyme were increased after the immobilization. PMID- 16245889 TI - [Quantitative detection of Vibrio parahaemolyticus by real-time TaqMan PCR]. AB - Vibrio parahaemolyticus is one of important human food pathogens. Traditional diagnostic tests for V. parahaemolyticus are laborious and always present false negative results. Therefore, it is important to develop a nucleic acid-based test for quantitative detection of V. parahaemolyticus. A TaqMan PCR assay was presented for quantitative detection of V. parahaemolyticus in pure cultures and oysters. The primers and probe were designed according to the gyrase B gene (gyrB) sequence of V. parahaemolyticus strains. Amplification of DNAs from 12 bacterial strains comprising 9 genera showed that all of the strains of V. parahaemolyticus tested (n = 4) were positive and all other species of strains tested (n = 8) were negative. The results of the TaqMan PCR with raw oysters inoculated with V. parahaemolyticus were comparable to those of pure cultures. The sensitivity of the assay was 1 CFU PCR Mixture(-1) and 10 CFU PCR Mixture(-1) in pure culture and inoculated raw oyster, respectively. The correlation rate was 0.99 (gamma2 = 0.99). The assay could be completed within 1h. The Real-time PCR can be used as a rapid screening tool for the presence of V. parahaemolyticus in seafood without prior isolation and characterization of the bacteria by traditional microbiological methods. PMID- 16245890 TI - [In vitro inhibitory efects on HBsAg and HBeAg secretion of 3 new components produced by Ganoderma lucidum in the medium contained Radix sophorae flavescentis extract]. AB - In order to enhance the medical effects of Ganoderma lucidum submergedly cultured broth, the aqueous extract of Radix sophorae flavescentis, a traditional Chinese medicine, was added into the cultivation medium of G. lucidum. The organic acids were extracted with ethanol, chloroform, 5% NaHCO3 and chloroform in turn from the cultured broth of Ganoderma lucidum which cultivation medium contained Radix sophorae flavescentis extract. Six new components were separated from the organic acids with preparative HPLC. Their inhibitory effects on HBsAg and HBeAg secretion of HBV DNA transferred HepG2 cell (2.2.15 cell) were investigated. The results indicate that 3 components of the six have significant inhibitory effects on the antigen secretion. PMID- 16245891 TI - [Burkholderia cepacia: our enemy or friend?]. AB - It was briefly described the beneficial and harmful effects of Burkholderia cepacia in agriculture, industry, medical science and environment protection since the bacterium was identified as causal organism of a crop in 1949. The hot points and problems in research and regulation of the bacterium were discussed and analyzed as well as the research suggestions in China. PMID- 16245892 TI - [Bacterial Fe(III) reduction]. AB - Bacterial Fe(III) reduction is an important pathway of bioenergy metabolism in the process of life evolution. Many kinds of archaebacteria and eubacteria are capable of reducing Fe(III) to conserve energy. Anaerobic Fe(III) respiration pathway involves many membranous proteins and regulating factors, especially the muti-haem c-type cytochromes are very important in the course of electron transportation. In addition, bacterial Fe(III) reduction play important roles in the biological geochemistry circulation and environmental remediation, therefore has vital environmental significance. PMID- 16245893 TI - [Advance in researches of drugs derived from marine bacteria]. AB - Marine bacteria are capable of producing a lot of unique bioactive substances, and therefore provide a luxuriant resource for screening new drugs. Bioactive substances derived from marine bacteria have bright prospect in marine drugs development and research. PMID- 16245894 TI - [Survey scheme of the main Chinese medicines resources]. AB - The survey of Chinese medicines resources concerns the development of the traditional Chinese medicine industry. It also directly influences the modernization process of traditional Chinese medicines. Owing to lacking of the scientific data on the Chinese medicines resources, it is a extremely urgent work to carry out survey of main Chinese medicine resources in nationwide. This paper explains the meaning and necessity. And it also investigated the survey object, contents, new technologies and methods. PMID- 16245895 TI - [Study on N, P and K absorption and accumulation in Ophiopogon japonicus]. AB - OBJECTIVE: To study the absorption and accumulation of N, P and K in Ophiopgon japonicus. METHOD: The contents of N, P, K and trace element were determined respectively by using the kjeldathl method digested with H2SO4 + H2O2, ammonium vanadstemolybdate chromoscopy, flame spectrometry and atomic absorption spectrophotometry. RESULT: The contents of N and K were the highest, and that of P was relatively lower O. japonicus. The changing pattern of N was low-high-low in the growth and development period. The accumulative speed of N had two relatively quicker and two relatively slower phases. The content of N is higher in nutritive organs; The changing pattern of P was similar to that of N, while the content of P is higher in leaves and nutritive roots at early stage of growth and development. It was high in root tuber during harvesting stage, and the accumulative quantity in the plant was increased gradually. The level of K was high in early stage and low in later stage of growth. The content of K was relatively higher in leaves and nutritive roots at early stage, and that in root tuber was slightly lower than the leaves but higher than that the nutritive roots from February to March. The accumulating quantity slightly increased in the plant. CONCLUSION: The application quantity of N should be increased gradually after transplant, stopped in autumn, and used again in early spring. Phosphate fertilizer is needed in autumn and winter, while large amount of potassium fertilizer shoule be used in winter tine. PMID- 16245896 TI - [Effects of plant growth substances on induction and culture of callus from Rhodiola quadrifida]. AB - OBJECTIVE: To investigate the effect of plant growth substances on induction and culture of callus from Rhodiola quadrifida and also to analyze salidroside contents in the callus. METHOD: The optimum combination of plant growth substances in MS solid medium for induction and culture of callus was established using orthogonal design. The contents of salidroside was analyzed by HPLC. RESULT: MS medium containing 2,4-D 1 mg x L(-1), NAA 2 mg x L(-1), 6-BA 0.5 mg x L(-1) and KT 0.1 mg x L(-1) could induce the callus from R. quadrifida most effectively;the induction rate was 83.3%. The optimized combination of plant growth substances for callus subculture was 2,4-D 1 mg x L(-1), 6-BA 0.1 mg x L( 1) and KT 0.5 mg x L(-1). The dry weight could reach 11.77 g x L(-1) when the callus was cultured in the optimum medium for 30 d and salidroside content was 0.28%. CONCLUSION: The quantities of plant growth substances required for induction and culture of callus are different in R. quadrifida. The callus could produce salidroside. PMID- 16245897 TI - [Study of comprehensive cultivation techniques of Pinellia ternate]. AB - OBJECTIVE: To study the effect of N, P, K and sowing norm on yield of Pinellia ternate. METHOD: The effects of N, P, K and sowing norm were studied by means of 4 factors and 3 levels orthogonal design experiments. RESULT AND CONCLUSION: The effects of the 4 factors decreased in following orders is sowing norm, N, K and P. Rational amounts of fertilizer and sowing norm are 48.0-96.0 kg x hm(-2) N, 90.0 kg x hm(-2) P, 90.0 kg x hm(-2) K, 3 000.0-3 750.0 kg x hm(-2). PMID- 16245898 TI - [Methodological studies on selectively removing toxins in Aristolochiae manshuriensis by chinese processing techniques]. AB - OBJECTIVE: To investigate the possibility of selectively detoxifying aristolochic acids in Aristolochiae manshuriensis (Guanmutong) by the chemical properties of aristolochic acids and traditional Chinese processing experience. METHOD: The technical parameters in processing technique of A. manshuriensis were optimized by orthogonal designed methods with aristolochic acid A. RESULT: The processing technique was soaked in 0.1 mol x L(-1) baking soda for several times and then processing with vinegar. Temperature was important factor to detoxify aristolochic acids. aristolochic acid A were removed over 90% from A. manshuriensis in laboratory and over 80% in medium-scale production by 10 batches from two origins of botanical drugs with different contents, and decreased to 0.35-0.60 mg x g(-1) in processed products. aristolochic acid A existed mostly salt-forms in the botanical drug. CONCLUSION: Most toxic components in Guanmutong could be removed by the new processing method. PMID- 16245899 TI - [Studies on hyaluronic acid as dendifier in Shuanghuanglian eye-drops]. AB - OBJECTIVE: To study the possibility of hyaluronic acid as densifier of Shuanguangliao eye-drops. METHOD: The factors related with hyaluronic acid s viscosity, such as pH-value and storing temperature, are tested in this experiment. At the same time, we checked the stimulation, stability of the densifier. RESULT: There was not effect on viscosity of pH-value and storing temperature. No stimulation on the eye was found after densified with hyaluronic acid. The viscosity properties of hyaluronic acid are stablile. CONCLUSION: The hyaliuronic acid added to Shuanghuanglian eye-drops are stabiliable and it can be applied in eye-drops. The increased viscosity is benefit to extend the residence time of drug in eye. PMID- 16245900 TI - [The preparation of arabinogalactan and the mechanism of release]. AB - OBJECTIVE: The study is the research on the preparation of arabinogalactan (AG) dropping pills and the releasing mechanism. METHOD: Use the orthogonal test to find out the best way to produce and advance the preparation of AG dropping pills, analysis according to the chart and DSC to find the releasing mechanism. RESULT: The best preparation conditions are: the liquid of AG is at 75 degrees C, the temperature above the polydimethls iloxane is 30 degrees C, the distance to the frizzed liquid is 6 cm, the speed of the liquid is 30 drop x min(-1). The chart and DSC suggest: The solid disoperation of AG-PREG 4000 the complex is in a certain form which made the melting point decreased obviously, so as to increase the solution of the medicine in carrier to increase the releasing speed. CONCLUSION: The best preparation is reasonable, AG and carrier become a form, the melting point is low, it can release fast. PMID- 16245901 TI - [The simultaneous determination of laetrile, paeoniflorin and paeonol in Jingzhi Guizhi Fuling capsule by HPLC]. AB - OBJECTIVE: To establish an HPLC method for the simultaneous determination of three major bioactive components in Jingzhi Guizhi Fuling capsules namely laetrile, paeoniflorin and paeonol. METHOD: A LiChrospher C18 column (4.6 mm x 250 mm, 5 microm) was used. The chromatography was carried out with a stepwise gradient programming. The mobile phase was acetonitrile-water (containing 0.1% phosphorous acid) and the flow rate was 1.0 mL x min. RESULT: The linear range of laetrile was 12.87-102.94 micron x mL(-1), r = 0.999 9, paeoniflorin 24.84 - 198.7 microg x mL(-1), r = 0.9999 and paeonol 12.57-100.56 microg x mL(-1), r = 0.999 9. The method is accurate with variation less than 1.5 % and recovery more than 95 %. CONCLUSION: The method was successfully applied to analyze three major bioactive components in Jingzhi Guizhi Fuling capsules. PMID- 16245902 TI - [Studies on chemical constituents in Flos Sophorae Carbonisatus]. AB - OBJECTIVE: To investigate the chemical constituents in Flos Sophorae Carbonisatus. METHOD: Silica gel column chromatography was used to separate and purify the chemical constituents. The structures were elucidated by spectral analysis. RESULT: Six compounds were isolated from Flos Sophorae Carbionisatus, and their structures were elucidated as maltol (1), 3-hydroxypyridine (2), malto 3-O-[6'-O-(4"-hydroxy-tans-cinnamoyl)-beta-D-glucopyranoside (3), 3-O-[beta-D galactopyranosyl-(1-->2)-beta-D-glucuronopyranosyl] sophoradiol ethyl ester (4), 3-O-[beta-D-galactopyranosyl-(1-->2)-beta-D-glucuronopyranosyl] sophoradiol methyl ester (5), rutin (6). CONCLUSION: 4 is a new compound, and 1,2,3,5 were first reported from Flos Sophorae Carbonisatus. PMID- 16245903 TI - [Chemical investingation of Chinese mangrove Sonneratia apetala II]. AB - OBJECTIVE: Isolation and structural elucidation of the constituents of China mangrove Sonneratia apetala. METHOD: chromatography methods were used for isolation of compounds, spectroscopic methods were used for structural identifyication. RESULT: seven known compounds named (+/-) symgaresinol, betulinic acid, lupeol, lupeone, stigmast-5-ene-3beta, 7alpha-diol, beta-alpha myrin hexadecaneate, physcoion were isolated. CONCLUSION: these known compouns were unreported previously from this plant. PMID- 16245904 TI - [Studies on chemical constituents in seeds of Taxus mairei II]. AB - OBJECTIVE: To study the chemical constituents in seeds of Taxus mairei. METHOD: Preparative HPLC, TLC and spectroscopic analyses were used to isolate and elucidate the chemical constituets in the plant. RESULT: Seven taxane diterpenoids were isolated from the seeds of T. mairei and identified as taxinine A(1), 9-deacetyltaxinine(2), 9-deacetyltaxinine E(3), 2-deacetyltaxinine(4), taxezopidine G(5), 2-deacetoxytaxinine J(6), 2-deacetoxytaxuspine C(7). CONCLUSION: Except compounds 5,6, all the compounds were obtained from seeds of this plant for the first time. PMID- 16245905 TI - [Studies on triterpenes chemical constituents in rhizome of Alisma gramineum]. AB - OBJECTIVE: To investigate the triterpenoids chemical constituents in rhizome of Alisma gramineum. METHOD: Silicon gel chromatography and HPLC techniques were employed for isolation and purification of the constituents, and the structures were elucidated by various spectral analyses such as NMR, MS and IR. RESULT: 5 protostane-type tetracyclic triterpenoids were isolated and identified as 16,23 oxidoalisol B(1), 11-deoxyalisol C(2), alisol F(3), 16beta-methoxyalisol B acetate (4), 16beta-hydroxyalisol B acetate (5). CONCLUSION: All of these compounds were found from A. gramineum for the first time. PMID- 16245906 TI - [Determination of two main components in bark of Paeonia suffruticosa by HPLC]. AB - OBJECTIVE: To establish a HPLC method for determination two constituents in bark of Paeonia Suffuticosa. METHOD: The reversed phase HPLC system consisting of an Alltima ODS column (4.6 mm x 150 mm, 5 microm) and a mixture of water-THF methanol-HAc (60:20:20:0.05) as the mobile phase was used. The flow rate was 0.8 mL x min(-1) and UV detection was set at 274 nm. RESULT: The assay displayed good linearity over the concentration ranges of 0.06-1.0 microg (r = 0.999 9, gallic acid) and 0.16-2.58 microg (r = 0.999 9, paeonol) respectively. The average recoveries (n = 9) of gallic acid and paeonol were 98.6% (RSD = 3.0%), 98.2% (RSD = 2.5%), respectively. The samples were extracted with methanol for 24 h bu maceration. CONCLUSION: The method is simple, accurate and can be used for the quality study of bark of P. suffruticosa. PMID- 16245907 TI - [Study on the chemical constituents of the volatile oil from aerial parts of Isodon eriocalyx var. laxiflora]. AB - OBJECTIVE: To study the chemical constituents of the volatile oil from the aerial parts of Isodon eriocalyx var. laxiflora. METHOD: The oil was obtained by hydrodistillation. The chemical compositions were separated and identified by GC MS. The relative contents in the oil were determined by area normalization. RESULT: 163 peaks were separated and 105 compounds were identified, constituting 85.68% of the total peak area. CONCLUSION: 105 compounds characterized by GC-MS analysis were found from I. eriocalyx var. laxiflora for the first time. PMID- 16245908 TI - [Experimental study of saponins from Tribulus terrestris on renal carcinoma cell line]. AB - OBJECTIVE: To investigate the effect of saponins from Tribulus terrestris (STT) on the renal carcinoma cell (786-0) in vitro, and inhibitory mechanisms. METHOD: Effects of SIT on the cytotoxicity, morphological changes of apoptosis, cell cycle and expression of Bcl-2 protein in the 786-0 were tested respectively by MTT method, Wright and acridine orange stain assay, as well as flow cytometry (FCM). RESULT: After the 786-0 was treated by STY, it was shown that: 1) A significant cytotoxic effect was observed by MTT assay; 2) Apoptosis-induced was viewed by Wright and acridine orange stain assay; 3) The distribution of 786-0 on S phase was increased; 4.) The expression of Bcl-2 protein and cyclin D1 was decreased. CONCLUSION: STT can significantly inhibit the growth of 786-0 in vitro, partially, by apoptosis. PMID- 16245909 TI - [Effect of Shuanghuang Shengbai granule on radiotherapy or chemotherapy induced leukopenia in mice]. AB - OBJECTIVE: To observe the effect of Shuanghuang Shengbai granule on mice leukopenia induced by ip cyclophosphamide (CTX) or radiation. METHOD: Mice leukopenia models were induced by ip CTX or radiation, and then treated with Shuanghuang Shengbai granule per oral. The peripheral hemogram, thymus index, spleen index, bone marrow nucleated cell (BMNC) and colony forming unit-spleen (CFU-S) were detected. The bone marrow cell differentiation was examined. The pathological slices of bone marrow were observed. RESULT: Shuanghuang Shengbai granule could increase the WBC, BMNC, CFU-S of model mice significantly; Shuanghuang Shengbai granule could make the granulocyte and erythrocyte index recovered to normal level and it could also protect the bone marrow hemotopoietic microenvironment from the harm of radiation. CONCLUSION: Shuanghuang Shengbai granule has apparent leukogenic function. PMID- 16245910 TI - [The effect of tetrandrine on the expression of the P170, LRP and TOPO II in S180's tumor cell induced by chemotherapy in the mice with acquired multi-drug resistance]. AB - OBJECTIVE: To observe the effect of tetrandrine on the P170 production expressed by multi-drug resistance gene, lung resistant protein (LRP), and topoisomeras II and elucidate the underlying molecular mechanism. METHOD: Cellular model of multi drug resistance was established in S180 tumor cell by means of the scheme of PFC chemotherapy at the dosage lower than that with curative effect. P170, LRP and TOPO II were measured by flow cytometry after the mouse model was treated with tetrandrine for 4 weeks. RESULT: tetrandrine obviously reduced the enhancement of express of P170, LRP and the activity of TOPO II in the tumor cells with multi drug resistance induced by chemotherapy. CONCLUSION: Tetrandrine significantly inhibits the multi-drug resistance of tumor cells induced by chemotherapy via diminishing both the expression of multi-drug resistance gene and the activity of topoisomeras II. PMID- 16245911 TI - [Effects of resveratrol on isolated thoracic aorta rings of rats]. AB - OBJECTIVE: To investigate the relaxative characteristics of resveratrol on thoracic aortic artery in the rat and its mechanism. METHOD: We perfused the isolated rings and observed the response of NA-induced artery contraction to resveratrol under the Ca2+-contained and Ca2+-free bath solutions. In the same way were the effect of reveratrol on the vascular smooth muscle observed by adding two different concentration of KCl (30 and 80 mmol x L(-1)), and the effect on the contraction of the vascular smooth muscle depending on the intracellular calcium and extracellular calcium were also observed by adding NA. We also observed the effect of resveratrol on the contraction of rings induced by NA in the presence of L-NNA and Glibenclamide. RESULT: Resveratrol relaxed rat aorta rings precontracted by NA in a dose-dependent manner. The relaxant effect of resveratrol on the rat rings of endothelium-denuded group was reduced compared with that of endothelium-intact group; the relaxant effect of resveratrol on rat rings was higher under the condition of Ca2+-free bath solution than that under the condition of Ca2+-contained bath solution. Resveratrol had a repressive effect on the aorta's contraction induced by intracellular calcium, but had no effect induced by extracellular calcium. Resveratrol relaxed the contractions induced by KCl 30 mmol x L(-1) as well as KCl 80 mmol x L(-1), but the contraction curve of KCl 80 mmol x L(-1) was shifted upward significantly. In the L-NNA group, the relaxant effect was attenuated by (26.0 +/- 4.6) %; but there was no change in the group of Glibenclamide ( P > 0.05). CONCLUSION: The results indicate that resveratrol relaxes vascular smooth muscle in an endothelium dependent manner. The mechanisms for this phenomenon seem to be related with promoting synthesis and release of NO, opening Ca2+ activated K+ channel (KCa channel) as well as the inhibition of Ca2+ influx and release of Ca2+ from intracellular stores. PMID- 16245913 TI - [Research progress in the reversion of traditional Chinese medicine on multidrug resistance of tumor]. AB - Researches on MDR (multidrug resistance) of tumor presently focus on seeking chemosensitizers with more targets, high efficiency and low toxicity from traditional Chinese medicine. This paper reviews the research progress in the reversion of MDR of leukemia, hepatocarcinoma, breast carcinoma and oral epithelioid neoplasia by TDM compound, its extracts, its groups of active ingredients or its active ingredients. PMID- 16245912 TI - [Dyuamical studies on metabolic chemistry of lignans from seeds of Arctium lappa]. AB - OBJECTIVE: To study the metabolic chemistry and pharmaco-dynamics characters of ligan from seeds of Arctium lappa. METHOD: HPLC method was used in the study. The analysis was carried out on C18 column. The mobile phase was CH3CN-0.05% H3PO4 (36:64) with flow-rate at 0.6 mL x min(-1) and wave-length of 210 nm. The column temperature was kept at 25 degrees C. RESULT: The results indicated that the ligan was detected in plasma and the main organs 5 min after po. The main metabolic production in plasma was arctigenin. In addition, arctigenin and an unknown product were found in metabolic production in the organs. CONCLUSION: The method was stable,simple and reproducible. It can be used to determine the metabolic product of the ligan. The metabolic chemistry of ligan in plasma was obviously different from that in the main organs. PMID- 16245914 TI - [Influence of Shenmai injection on blood serum tumor necrosis factor and bone marrow CD34+ cell's apoptosis of chronic aplastic anemia patient]. AB - OBJECTIVE: To investigate the effect of Shenmai injection on chronic aplastic anemia patients and its mechanism. METHOD: Sixty-five chronic aplastic anemia patients were randomized into treatment group and control group. The patients of the treatment group were treated by injecting Shenmai injection and taking western medicine orally, those of the control group taking western medicine orally only, then the effect was evaluated. The concentration of tumor necrosis factor-alpha (TNF-alpha) in blood serum was detected and the apoptosis of bone marrow CD34+ cell was analysed by DNA ISEL technic before and after treatment. RESULT: The effective rate of the treatment group and the control group was 63.6 % and 40.6 % respectively, the effect of the Shenmai injection on the treatment group was obviously better than that of the control group (P < 0.01). Before treatment, the concentration of TNF-alpha in blood serum and the apoptosis rate of bone marrow CD34+ cell of the chronic aplastic anemia patient were higher than normal (P < 0.01). After treatment, the concentration of TNF-alpha in blood serum of the treatment group decreased obviously (P < 0.01), and the apoptosis rate of bone marrow CD34+ cell of the treatment group also decreased (P < 0.05), which had significant difference compared with those of the control group (P < 0.05). CONCLUSION: Shenmai injection is efficient to chronic aplastic anemia. The mechanism is decreasing the concentration of TNF-alpha in blood serum and the apoptosis rate of bone marrow CD34+ cell. PMID- 16245915 TI - [Infectious endocarditis]. PMID- 16245916 TI - [What is your diagnosis? Lung aplasia on the left]. PMID- 16245917 TI - [Outpatient alcohol withdrawal]. AB - Indications and contraindications for an alcohol withdrawal treatment of outpatients are presented, with particular reference to the symptoms and the diagnosis of the withdrawal syndrome. The general conditions and the practical process of outpatient detoxification are outlined. At the same time the question of medication of the withdrawal syndrome is addressed. PMID- 16245918 TI - [Molecular signatures in ovarian carcinoma]. PMID- 16245919 TI - [Swollen leg with blisters]. AB - We report the case of a 84-year-old woman suffering from strong pain in her right leg initially resembling thrombosis of deep veins. Eight hours after admission a superficial blister developed at the calf with following hemorrhagic aspect and spontanous eruption of clear yellowish fluid. Later on a new blister appeared at the thigh. The patient died 33 hours after admission of streptococcal toxic shock syndrome. The latter was based on a necrotizing fasciitis. Streptoccus pyogenes (group A) could be cultivated from the blood and fluid of the blister. We discuss the clinical presentation of necrotizing fasciitis with "pain out of proportion" as characteristic complaint and the appropriate management. PMID- 16245920 TI - Directed evolution of highly homologous proteins with different folds by phage display: implications for the protein folding code. AB - To better understand how amino acid sequences specify unique tertiary folds, we have used random mutagenesis and phage display selection to evolve proteins with a high degree of sequence identity but different tertiary structures (homologous heteromorphs). The starting proteins in this evolutionary process were the IgG binding domains of streptococcal protein G (G(B)) and staphylococcal protein A (A(B)). These nonhomologous domains are similar in size and function but have different folds. G(B) has an alpha/beta fold, and A(B) is a three-helix bundle (3 alpha). IgG binding function is used to select for mutant proteins which retain the correct tertiary structure as the level of sequence identity is increased. A detailed thermodynamic analysis of the folding reactions and binding reactions for a pair of homologous heteromorphs (59% identical) is presented. High resolution NMR structures of the pair are presented by He et al. [(2005) Biochemistry 44, 14055-14061]. Because the homologous but heteromorphic proteins are identical at most positions in their sequence, their essential folding signals must reside in the positions of nonidentity. Further, the thermodynamic linkage between folding and binding is used to assess the propensity of one sequence to adopt two unique folds. PMID- 16245921 TI - Solution NMR structures of IgG binding domains with artificially evolved high levels of sequence identity but different folds. AB - We describe here the solution NMR structures of two IgG binding domains with highly homologous sequences but different three-dimensional structures. The proteins, G311 and A219, are derived from the IgG binding domains of their wild type counterparts, protein G and protein A, respectively. Through a series of site-directed mutations and phage display selections, the sequences of G311 and A219 were designed to converge to a point of high-level sequence identity while keeping their respective wild-type tertiary folds. Structures of both artificially evolved sequences were determined by NMR spectroscopy. The main chain fold of G311 can be superimposed on the wild-type alpha/beta protein G structure with a backbone rmsd of 1.4 A, and the A219 structure can be overlaid on the wild-type three-alpha-helix protein A fold also with a backbone rmsd of 1.4 A. The structure of G311, in particular, accommodates a large number of mutational changes without undergoing a change in the overall fold of the main chain. The structural differences are maintained despite a high level (59%) of sequence identity. These proteins serve as starting points for further experiments that will probe basic concepts of protein folding and conformational switching. PMID- 16245922 TI - Role of electrostatics and salt bridges in stabilizing the compound I radical in ascorbate peroxidase. AB - Cytochrome c (CcP) and ascorbate peroxidase (APX) are heme peroxidases which have very similar active site structures yet differ substantially in the properties of compound I, the intermediate formed upon reaction with peroxides. Although both peroxidases have a tryptophan in the proximal heme pocket, Trp191 in CcP and Trp179 in APX, only Trp191 in CcP forms a stable cation radical while APX forms the more traditional porphyrin pi-cation radical. Previous work [Barrows, T. P., et al. (2004)Biochemistry 43, 8826-8834] has shown that converting three methionine residues in the cytochrome c peroxidase (CcP) proximal heme pocket to the corresponding residues in APX dramatically decreased the stability of the Trp191 radical in CcP compound I. On the basis of these results, we reasoned that replacing the analogous residues at positions 160, 203, and 204 in APX with methionine should stabilize a Trp179 radical in APX compound I. Steady- and transient-state kinetics of this mutant (designated APX3M) show a significant destabilization of the native porphyrin pi-radical, while electron paramagnetic resonance (EPR) studies show an increase in the intensity of the signal at g = 2.006 with characteristics consistent with formation of a Trp radical. This hypothesis was tested by replacing Trp179 with Phe in the APX3M background. The EPR spectrum of this mutant was very similar to that of the CcP W191G mutant which is known to form a tyrosine radical. Previously published theoretical studies [Guallar, V., et al. (2003) Proc. Natl. Acad. Sci. U.S.A. 100, 6998-7002] suggest that electrostatic shielding of the heme propionates also plays a role in the stability of the porphyrin radical. Arg172 in APX hydrogen bonds with one of the heme propionates. Replacing Arg172 with an asparagine residue in the APX3M background generates a mutant which no longer forms the full complement of the compound I porphyrin pi-radical. These results suggest that the electrostatics of the proximal pocket and the shielding of propionate groups by salt bridges are critical factors controlling the location of a stable compound I radical in heme peroxidases. PMID- 16245923 TI - The structure-activity relationship of ferric pyoverdine bound to its outer membrane transporter: implications for the mechanism of iron uptake. AB - Under iron limitation, Pseudomonas aeruginosa ATCC 15692 secretes a major siderophore, pyoverdine I (PvdI). This molecule chelates iron in the extracellular medium and shuttles it into the cells via a specific outer membrane transporter, FpvAI. PvdI consists of a fluorescent chromophore derived from 2,3 diamino-6,7-dihydroxyquinoline and containing one of the bidentate groups involved in iron chelation, linked to a peptide moiety containing the two other bidentate groups required for binding to Fe(3+). Kinetic studies, based on the fluorescence properties of this siderophore, showed that pH 8.0 was optimal for the binding of PvdI and PvdI-Fe to FpvAI. We investigated the mechanism of interaction of PvdI and PvdI-Fe with FpvAI, by synthesizing various analogues of this siderophore, determining their affinity for FpvAI in vitro and in vivo and their ability to transport iron, and interpreting the results obtained in light of the structure of FpvAI-PvdI. Our findings demonstrate that the succinyl moiety linked to the chromophore of PvdI and the first amino acid of the peptide moiety can be sterically hindered with no effect on binding or the iron uptake properties of PvdI-Fe. Moreover, the sequence and the structure of the peptide moiety of PvdI seems to be more important for the iron uptake step than for the binding of the siderophore to FpvAI. Finally, the efficiency of iron uptake and of recycling of the various PvdI analogues after iron release suggests that iron dissociates from PvdI on FpvAI or in the periplasm. All these data have serious implications for the specificity and mechanism of PvdI-mediated iron transport in P. aeruginosa. PMID- 16245924 TI - Exploring the mechanism of binding of UDP-galactopyranose to UDP-galactopyranose mutase by STD-NMR spectroscopy and molecular modeling. AB - UDP-galactopyranose mutase (UGM) is the key enzyme involved in the biosynthesis of Galf. In this study, reliable structural binding modes of the natural substrate, UDP-Galp, and inhibitor, UDP, in the UGM active site were provided with the combined use of STD-NMR spectroscopy, molecular modeling, and CORCEMA-ST calculations. UDP-Galp and UDP exhibited similar binding epitopes recognized by UGM. However, the relative binding affinities of the ligands changed dramatically upon reduction of UGM, as explored by competitive STD-NMR experiments. UDP-Galp competes with UDP for binding to UGM, especially when UGM is in its reduced state. Docking studies for predicting the binding mode within the active site of the two monomers in UGM explored the possibility that the mobile loop might act as a gateway for substrate binding, and the structure of the binding cleft in monomer A might be a closer approximation of the substrate-bound active site than monomer B. Important information regarding the critical interactions of UGM with UDP-Galp has been obtained. PMID- 16245925 TI - JNK-interacting leucine zipper protein is a novel scaffolding protein in the Galpha13 signaling pathway. AB - Scaffolding proteins play a critical role in conferring specificity and fidelity to signaling pathways. The JNK-interacting leucine zipper protein (JLP) has been identified as a scaffolding protein involved in linking components of the JNK signaling module. Galpha(12) and Galpha(13), the alpha-subunits of heterotrimeric G proteins G12 and G13, respectively, stimulate the JNK module in diverse cell types. Here, we report that Galpha(13) physically interacts with JLP, and this interaction enhances Galpha(13)-mediated JNK activation. We also demonstrate endogenous interaction between JLP and Galpha(13) in MCF-7 cells. JLP interaction is specific to the G12 family of alpha-subunits via its C-terminal domain (termed GID-JLP), spanning amino acids 1165-1307, and this interaction is more pronounced with the mutationally or functionally activated form of Galpha(13) compared to that of wild-type Galpha(13). The presence of a ternary complex consisting of Galpha(13), JLP, and JNK suggests a role for JLP in tethering Galpha(13) to the signaling components involved in JNK activation. Coexpression of GID-JLP disrupts ternary complex formation in addition to attenuating Galpha(13)-stimulated JNK activity. These findings identify JLP as a novel scaffolding protein in the Galpha(13)-mediated JNK signaling pathway. PMID- 16245926 TI - Role of Ca2+ in the electrostatic stability and the functional activity of the globular domain of human C1q. AB - C1q is the recognition subunit of the classical pathway of the complement system and a major connecting link between classical pathway-driven innate immunity and IgG- or IgM-mediated acquired immunity. The basic structural subunit of C1q is composed of an N-terminal triple-helical collagen-like region and a C-terminal heterotrimeric globular head domain (gC1q) that is made up of individual A, B, and C chains. Recent crystallographic studies have revealed that the gC1q domain, which is the main target-binding region of C1q, has a compact and spherical heterotrimeric assembly, held together by both electrostatic and nonpolar interactions, with quasi-3-fold symmetry. A characteristic feature of the gC1q domain is the presence of a exposed Ca(2+) located near the apex. We have investigated, using theoretical and experimental approaches, the role of Ca(2+) in the electrostatic stability and target-binding properties of the native C1q as well as recombinant monomeric forms of the C-terminal regions of the A, B, and C chains. Here, we report that Ca(2+) primarily influences the target recognition properties of C1q toward IgG, IgM, C-reactive protein, and pentraxin 3. At pH 7.4, the loss of Ca(2+) leads to changes in the direction of electric moment from coaxial (where the putative C-reactive protein-binding site is located) to perpendicular to the molecular axis (toward the most likely IgG-binding site), which appears important for target recognition by C1q and subsequent complement activation. PMID- 16245927 TI - Regulation of the wild-type and Y1235D mutant Met kinase activation. AB - Met receptor tyrosine kinase plays a crucial role in the regulation of a large number of cellular processes and, when deregulated by overexpression or mutations, leads to tumor growth and invasion. The Y1235D mutation identified in metastases was shown to induce constitutive activation and a motile-invasive phenotype on transduced carcinoma cells. Wild-type Met activation requires phosphorylation of both Y1234 and Y1235 in the activation loop. We mapped the major phosphorylation sites in the kinase domain of a recombinant Met protein and identified the known residues Y1234 and Y1235 as well as a new phosphorylation site at Y1194 in the hinge region. Combining activating and silencing mutations at these sites, we characterized in depth the mechanism of activation of wild type and mutant Met proteins. We found that the phosphotyrosine mimetic mutation Y1235D is sufficient to confer constitutive kinase activity, which is not influenced by phosphorylation at Y1234. However, the specific activity of this mutant was lower than that observed for fully activated wild-type Met and induced less phosphorylation of Y1349 in the signaling site, indicating that this mutation cannot entirely compensate for a phosphorylated tyrosine at this position. The Y1194F silencing mutation yielded an enzyme that could be activated to a similar extent as the wild type but with significantly slower activation kinetics, underlying the importance of this residue, which is conserved among different tyrosine kinase receptors. Finally, we observed different interactions of wild-type and mutant Met with the inhibitor K252a that may have therapeutic implications for the selective inhibition of this kinase. PMID- 16245928 TI - A Ca2+/calmodulin-binding peroxidase from Euphorbia latex: novel aspects of calcium-hydrogen peroxide cross-talk in the regulation of plant defenses. AB - Calmodulin (CaM) is a ubiquitous Ca(2+) sensor found in all eukaryotes, where it participates in the regulation of diverse calcium-dependent physiological processes. In response to fluctuations of the intracellular concentration of Ca(2+), CaM binds to a set of unrelated target proteins and modulates their activity. In plants, a growing number of CaM-binding proteins have been identified that apparently do not have a counterpart in animals. Some of these plant-specific Ca(2+)/CaM-activated proteins are known to tune the interaction between calcium and H(2)O(2) in orchestrating plant defenses against biotic and abiotic stresses. We previously characterized a calcium-dependent peroxidase isolated from the latex of the Mediterranean shrub Euphorbia characias (ELP) [Medda et al. (2003) Biochemistry 42, 8909-8918]. Here we report the cDNA nucleotide sequence of Euphorbia latex peroxidase, showing that the derived protein has two distinct amino acid sequences recognized as CaM-binding sites. The cDNA encoding for an E. characias CaM was also found and sequenced, and its protein product was detected in the latex. Results obtained from different CaM binding assays and the determination of steady-state parameters showed unequivocally that ELP is a CaM-binding protein activated by the Ca(2+)/CaM system. To the best of our knowledge, this is the first example of a peroxidase regulated by this classic signal transduction mechanism. These findings suggest that peroxidase might be another node in the Ca(2+)/H(2)O(2)-mediated plant defense system, having both positive and negative effects in regulating H(2)O(2) homeostasis. PMID- 16245929 TI - Low-temperature interquinone electron transfer in photosynthetic reaction centers from Rhodobacter sphaeroides and Blastochloris viridis: characterization of Q(B)- states by high-frequency electron paramagnetic resonance (EPR) and electron nuclear double resonance (ENDOR). AB - High-frequency electron paramagnetic resonance (HF EPR) techniques have been employed to look for localized light-induced conformational changes in the protein environments around the reduced secondary quinone acceptor (Q(B)(-)) in Rhodobacter sphaeroides and Blastochloris viridis RCs. The Q(A)(-) and Q(B)(-) radical species in Fe-removed/Zn-replaced protonated RCs substituted with deuterated quinones are distinguishable with pulsed D-band (130 GHz) EPR and provide native probes of both the low-temperature Q(A)(-)Q(B) --> Q(A)Q(B)(-) electron-transfer event and the structure of trapped conformational substates. We report here the first spectroscopic evidence that cryogenically trapped, light induced changes enable low-temperature Q(A)(-)Q(B) --> Q(A)Q(B)(-) electron transfer in the B. viridis RC and the first observation of an inactive, trapped P(+)Q(B)(-) state in both R. sphaeroides and B. viridis RCs that does not recombine at 20 K. The high resolution and orientational selectivity of HF electron-nuclear double resonance (ENDOR) allows us to directly probe protein environments around Q(B)(-) for distinct P(+)Q(B)(-) kinetic RC states by spectrally selecting specific nuclei in isotopically labeled samples. No structural differences in the protein structure near Q(B)(-) or reorientation (within 5 degrees ) of Q(B)(-) was observed with HF ENDOR spectra of two states of P(+)Q(B)(-): "active" and "inactive" states with regards to low-temperature electron transfer. These results reveal a remarkably enforced local protein environment for Q(B) in its reduced semiquinone state and suggest that the conformational change that controls reactivity resides beyond the Q(B) local environment. PMID- 16245930 TI - Cooperative binding of midazolam with testosterone and alpha-naphthoflavone within the CYP3A4 active site: a NMR T1 paramagnetic relaxation study. AB - Recent studies have indicated that CYP3A4 exhibits non-Michaelis-Menten kinetics for numerous substrates. Both homo- and heterotropic activation have been reported, and kinetic models have suggested multiple substrates within the active site. We provide some of the first physicochemical data supporting the hypothesis of allosteric substrate binding within the CYP3A4 active site. Midazolam (MDZ) is metabolized by CYP3A4 to two hydroxylated metabolites, 1'- and 4 hydroxymidazolam. Incubations using purified CYP3A4 and MDZ showed that both alpha-naphthoflavone (alpha-NF) and testosterone affect the ratio of formation rates of 1'- and 4-hydroxymidazolam. Similar to previous reports, alpha-NF was found to promote formation of 1'-hydroxymidazolam, while testosterone stimulated formation of 4-hydroxymidazolam. NMR was used to measure the closest approach of individual MDZ protons to the paramagnetic heme iron of CYP3A4 using paramagnetic T(1) relaxation measurements. Solutions of 0.2 microM CYP3A4 with 500 microM MDZ resulted in calculated distances between 7.4 and 8.3 A for all monitored MDZ protons. The distances were statistically equivalent for all protons except C3-H and were consistent with the rotation within the active site or sliding parallel to the heme plane. When 50 microM alpha-NF was added, proton-heme iron distances ranged from 7.3 to 10.0 A. Consistent with kinetics of activation, the 1' position was situated closest to the heme, while the fluorophenyl 5-H proton was the furthest. Proton-heme iron distances for MDZ with CYP3A4 and 50 microM testosterone ranged from 7.7 to 9.0 A, with the flourophenyl 5-H proton furthest from the heme iron and the C4-H closest to the heme, also consistent with kinetic observations. When titrated with CYP3A4 in the presence of MDZ, testosterone and alpha-NF resonances themselves exhibited significant broadening and enhanced relaxation rates, indicating that these effector molecules were also bound within the CYP3A4 active site near the paramagnetic heme iron. These results suggest that the effector exerts its cooperative effects on MDZ metabolism through simultaneous binding of MDZ and effector near the CYP3A4 heme. PMID- 16245931 TI - Eosinophil-granule major basic protein, a C-type lectin, binds heparin. AB - The eosinophil major basic protein (EMBP), a constituent of the eosinophil secondary granule, is implicated in cytotoxicity and mediation of allergic disorders such as asthma. It is a member of the C-type lectin family, but lacks a Ca(2+)- and carbohydrate-binding site as seen in other members of this family. Here, we report the crystal structure of EMBP in complex with a heparin disaccharide and in the absence of Ca(2+), the first such report of any C-lectin with this sugar. We also provide direct evidence of binding of EMBP to heparin and heparin disaccharide by surface plasmon resonance. We propose that the sugars recognized by EMBP are likely to be proteoglycans such as heparin, leading to new interpretations for EMBP function. PMID- 16245932 TI - Chemoenzymatic approaches for streamlined detection of active site modifications on thiotemplate assembly lines using mass spectrometry. AB - For the direct interrogation of peptides harboring covalently modified serines in nonribosomal peptide synthetases, streamlined methodologies described here employ proteolysis and reporter-coenzyme A analogues of four types. The chromophoric and fluorescent coenzyme A analogues pyrene-maleimidyl-S-CoA and BODIPY-FL-N-(2 aminoethyl)maleimidyl-S-CoA were enzymatically loaded onto the active site serines harbored in the ArCP, PCP1, and PCP2 thiolation domains of PchE and PchF, the nonribosomal peptide synthetases responsible for the biosynthesis of the siderophore pyochelin. During the chromatographic separation of cyanogen bromide digests, observation of the absorbance (at 338 and 504 nm) or fluorescence (after irradiation at 365 nm) enabled the selective detection of peptides containing each active site serine. This resulted in quick detection of each active site peptide by Fourier transform mass spectrometry in the fully reconstituted pyochelin system. The loading of short acyl chain reporters in equimolar quantities permitted further insights into digestion heterogeneity and side reactions by virtue of a mass shift signature on each active site peptide. The chromatographic shift of the reporter-loaded peptides relative to peptides carrying on pathway intermediates was 2 min at 7 kDa, providing a general strategy for efficient localization of "carrier" peptides in complex digests of thiotemplate enzymes. Also, the use of the affinity reporter, biotin-maleimidyl-S coenzyme A, permitted the isolation of intact synthetases at high purity via removal of contaminating Escherichia coli proteins. PMID- 16245933 TI - The preferred conformation of the tripeptide Ala-Phe-Ala in water is an inverse gamma-turn: implications for protein folding and drug design. AB - Recent studies have provided evidence that peptides as short as tripeptides do adopt preferred conformations. Here we report that the tripeptide Ala-Phe-Ala (AFA) in aqueous solution preferentially forms an inverse gamma-turn. Circular dichroism (CD) indicated the presence of a predominant turn structure, and Fourier transform infrared (FTIR) bands suggested the presence of a gamma-turn forming a bifurcated H-bond with the solvent molecules. The high-resolution structure was obtained by a combined use of NMR spectroscopy and calculations. On the basis of 30 unambiguous ROESY-derived distance restraints (including the Halpha-NH NOE between Ala(1) and Ala(3) and a hydrogen bond between the CO group of Ala(1) and the NH group of Ala(3)), calculations clearly demonstrated the presence of an inverse gamma-turn centered on Phe(2). From NOE data, we estimated a mole fraction for the gamma-turn of 0.65. Since for AFA an extended beta-strand was also reported [Eker, F., Griebenow, K., Cao, X., Nafie, L. A., and Schweitzer Stenner, R. (2004) Proc. Natl. Acad. Sci. U.S.A. 101, 10054-10059], we investigated the possibility that gamma-turn and beta-strand may represent two major conformations. By using a best-fit procedure that calculated experimental NOEs as weighted averages of the effects originating from both structures, we were able to calculate with good accuracy the backbone NOEs at 280 K in terms of the two limiting conformers, yielding a mole fraction for the gamma-turn and beta strand conformations of 0.60 and 0.40, respectively, in good agreement with those found by NOE data. The implication of the existence of a preferred conformation by a small structural element is discussed in the context of the nucleation of protein folding events and the design of small peptide and peptidomimetic drugs. PMID- 16245934 TI - Insight into the roles of tyrosine 82 and glycine 253 in the Escherichia coli adenine glycosylase MutY. AB - The oxidation product of 2'-deoxyguanosine, 7,8-dihydro-8-oxo-2'-deoxyguanosine (OG), produces G:C to T:A transversion mutations. The Escherichia coli base excision repair glycosylase MutY plays an important role in preventing OG associated mutations by removing adenines misincorporated opposite OG lesions during DNA replication. Recently, biallelic mutations in the human MutY homologue (hMYH) have been correlated with the development of colorectal cancer. The two most common mutations correspond to two single amino acid substitutions in the hMYH protein: Y165C and G382D [Al-Tassan, N., et al. (2002) Nat. Genet. 30, 227 232]. Previously, our laboratory analyzed the adenine glycosylase activity of the homologous variant E. coli MutY enzymes, Y82C and G253D [Chmiel, N. H., et al. (2003) J. Mol. Biol. 327, 431-443]. This work demonstrated that both variants have a reduced adenine glycosylase activity and affinity for substrate analogues compared to wild-type MutY. Recent structural work on Bacillus stearothermophilus MutY bound to an OG:A mismatch-containing duplex indicates that both residues aid in recognition of OG [Fromme, J. C., et al. (2004) Nature 427, 652-656]. To determine the extent with which Tyr 82 and Gly 253 contribute to catalysis of adenine removal by E. coli MutY, we made a series of additional modifications in these residues, namely, Y82F, Y82L, and G253A. When the substrate analogue 2' deoxy-2'-fluoroadenosine (FA) in duplex paired with G or OG is used, both Y82F and G253A showed reduced binding affinity, and G253A was unable to discriminate between OG and G when paired with FA. Additionally, compromised glycosylase activity of Y82F, Y82C, and G253A MutY was observed using the nonoptimal G:A substrate, or at low reaction temperatures. Interestingly, adenine removal from an OG:A-containing DNA substrate by Y82C MutY was also shown to be extremely sensitive to the NaCl concentration. The most surprising result was the remarkably similar activity of Y82L MutY to the WT enzyme under all conditions examined, indicating that a leucine residue may effectively replace tyrosine for intercalation at the OG:A mismatch. The results contained herein provide further insight regarding the intricate roles of Tyr 82 and Gly 253 in the OG recognition and adenine excision functions of MutY. PMID- 16245935 TI - Telomerase as a DNA-dependent DNA polymerase. AB - Telomerase is a specialized reverse transcriptase, which catalyzes the addition of telomeric repeats to the 3' ends of linear chromosomes using its integral RNA subunit as the template. An active Tetrahymena thermophila telomerase complex can be reconstituted in vitro from two essential components, tTERT, the catalytic protein subunit, and tTR, the RNA subunit. While the sequence specificity of telomerase has been investigated using template sequence mutants, there is no information regarding its backbone specificity. To address this question, we engineered two mutant forms of the telomerase RNA subunit that contain DNA only in the templating region and used rabbit reticulocyte lysates to reconstitute telomerase activity with the chimeric tTRs. The resultant telomerase mutants were able to extend telomeric DNA primers, albeit with reduced efficiency compared to the wild type. The reduced activity is presumed to be a function of the nascent DNA-template duplex structure. Additionally, the DNA-dependent telomerase mutants were RNase-sensitive, confirming that nontemplate portions of tTR are critical for maintaining activity of the telomerase ribonucleoprotein complex even after it is assembled. The splint ligation approach that we outline will allow the generation of tTR mutants containing a variety of nucleotide analogues, facilitating more elaborate studies of the interactions between the telomerase template and active site. PMID- 16245936 TI - Stability and DNA binding ability of the DNA binding domains of interferon regulatory factors 1 and 3. AB - The thermodynamic properties and DNA binding ability of the N-terminal DNA binding domains of interferon regulatory factors IRF-1 (DBD1) and IRF-3 (DBD3) were studied using microcalorimetric and optical methods. DBD3 is significantly more stable than DBD1: at 20 degrees C the Gibbs energy of unfolding of DBD3 is 28.6 kJ/mol, which is 2 times larger than that of DBD1, -14.9 kJ/mol. Fluorescence anisotropy titration experiments showed that at this temperature the association constants with the PRDI binding site are 1.1 x 10(6) M(-)(1) for DBD1 and 3.6 x 10(6) M(-)(1) for DBD3, corresponding to Gibbs energies of association of -34 and -37 kJ/mol, respectively. However, the larger binding energy of DBD3 is due to its larger electrostatic component, while its nonelectrostatic component is smaller than that of DBD1. Therefore, DBD1 appears to have more sequence specificity than DBD3. Binding of DBD1 to target DNA is characterized by a substantially larger negative enthalpy than binding of DBD3, implying that the more flexible structure of DBD1 forms tighter contacts with DNA than the more rigid structure of DBD3. Thus, the strength of the DBDs' specific association with DNA is inversely related to the stability of the free DBDs. PMID- 16245937 TI - Mapping peptides correlated with transmission of intrasteric inhibition and allosteric activation in human cystathionine beta-synthase. AB - Cystathionine beta-synthase plays a key role in the intracellular disposal of homocysteine and is the single most common locus of mutations associated with homocystinuria. Elevated levels of homocysteine are correlated with heart disease, Alzheimer's and Parkinson's diseases, and neural tube defects. Cystathionine beta-synthase is modular and subjected to complex regulation, but insights into the structural basis of this regulation are lacking. We have employed hydrogen exchange mass spectrometry to map peptides whose motions are correlated with transmission of intrasteric inhibition and allosteric activation. The mass spectrometric data provide an excellent correlation between kinetically and conformationally distinguishable states of the enzyme. We also demonstrate that a pathogenic regulatory domain mutant, D444N, is conformationally locked in one of two states sampled by the wild type enzyme. Our hydrogen exchange data identify surfaces that are potentially involved in the juxtaposition of the regulatory and catalytic domains and form the basis of a docked structural model for the full-length enzyme. PMID- 16245938 TI - Solution structure, stability, and nucleic acid binding of the hyperthermophile protein Sso10b2. AB - The Sso10b (or Alba) family of proteins is a conserved group of archaeal and eukaryotic proteins which are thought to play a role in both chromatin organization and RNA metabolism. We describe here the solution structure and properties of Sso10b2 from Sulfolobus solfataricus. NMR data including residual dipolar couplings and (15)N relaxation data demonstrated that the protein adopts a beta(1)alpha(1)beta(2)alpha(2)beta(3)beta(4) topology with an IF-3-like fold. The protein dimerizes in solution at 30 degrees C via a hydrophobic surface defined by the C-terminal alpha(2)beta(3)beta(4) elements with a structure similar to one of the putative dimers indicated by previous crystal structures. DSC and circular dichroism data demonstrated an unusual two-state structural transition near the growth temperature which led to an increase in beta-sheet content without dissociation of the dimer. The cooperativity of the transition exceeded that of a dimer at pH 7, demonstrating the presence of higher order oligomers near the growth temperature at pH 7. Reverse titrations of Sso10b2 with nucleic acid showed that the protein binds single-stranded DNA (K(d) of 3 x 10( )(7) M) with higher affinity than RNA (1.3 x 10(-)(6) M) or double-stranded DNA (1.5 x 10(-)(5) M) in 10 mM KH(2)PO(4) (pH 7.0, 20 degrees C). NMR chemical shift perturbation data indicated that single-stranded DNA and RNA binding occurred across the same dimer interface and encompassed a surface defined by the C terminal ends of the beta(1), beta(2), and beta(3) strands of each monomer. PMID- 16245939 TI - Effect of anions on the photocycle of halorhodopsin. Substitution of chloride with formate anion. AB - Halorhodopsin from Natronomonas pharaonis is a light-driven chloride pump which transports a chloride anion across the plasma membrane following light absorption by a retinal chromophore which initiates a photocycle. It was shown that the chloride anion bound in the vicinity of retinal PSB can be replaced by several inorganic anions, including azide which converts the chloride pump into a proton pump and induces formation of an M-like intermediate detected in the bR photocycle but not in native halorhodopsin. Here we have studied the possibility of replacing the chloride anion with organic anions and have followed the photocycle under several conditions. It is revealed that the chloride can be replaced with a formate anion but not with larger organic anions such as acetate. Flash photolysis experiments detected in the formate pigment an M-like intermediate characterized by a lifetime much longer than that of the O intermediate. The lifetime of the M-like intermediate depends on the pH, and its decay is significantly accelerated at low pH. The decay rate exhibited a titration-like curve, suggesting that the protonation of a protein residue controls the rate of M decay. Similar behavior was detected in N. pharaonis pigments in which the chloride anion was replaced with NO(2)(-) and OCN(-) anions. It is suggested that the formation of the M-like intermediate indicates branching pathways from the L intermediate or basic heterogeneity in the original pigment. PMID- 16245940 TI - The two Caenorhabditis elegans actin-depolymerizing factor/cofilin proteins differently enhance actin filament severing and depolymerization. AB - Actin-depolymerizing factor (ADF)/cofilin enhances the turnover of actin filaments by two separable activities: filament severing and pointed-end depolymerization. Multicellular organisms express multiple ADF/cofilin isoforms in a tissue-specific manner, and the vertebrate proteins are grouped into ADFs and cofilins on the basis of their biochemical activity. A recent comparative study has shown that ADF has greater severing and depolymerizing activities than cofilin [Chen, H., Bernstein, B. W., Sneider, J. M., Boyle, J. A., Minamide, L. S., and Bamburg, J. R. (2004) Biochemistry 43, 7127-7142]. Here, we show that the two Caenorhabditis elegans ADF/cofilin isoforms exhibit different activities for severing and depolymerizing actin filaments. The ADF-like non-muscle isoform UNC 60A had greater activities to cause net depolymerization and inhibit polymerization than the cofilin-like muscle isoform UNC-60B. Surprisingly, UNC 60B exhibited much stronger severing activity than UNC-60A, which was the opposite of what was observed for vertebrate counterparts. Moreover, UNC-60B induced much faster pointed-end depolymerization of rabbit muscle actin than UNC 60A, while UNC-60A caused slightly faster depolymerization of C. elegans actin than UNC-60B. These results suggest that cofilin-like UNC-60B is kinetically more efficient in enhancing actin turnover than ADF-like UNC-60A, while ADF-like UNC 60A is suitable for maintaining higher concentrations of monomeric actin. These functional differences might be specifically adapted for different actin dynamics in muscle and non-muscle cells. PMID- 16245941 TI - Ionization of His 55 at the dimer interface of dynein light-chain LC8 is coupled to dimer dissociation. AB - LC8 is a highly conserved light-chain subunit of cytoplasmic dynein that interacts with a wide variety of cellular proteins and is presumed to play a fundamental role in dynein assembly and cargo recruitment and in the assembly of protein complexes unrelated to dynein. LC8 is a dimer at physiological pH but dissociates to a folded monomer at pH < 4.8. We have suggested that acid-induced dimer dissociation is due to protonation of His 55, which is stacked against His 55' and completely buried in the dimer interface. In this work, we show that the pH-induced dissociation is reversible and indeed governed by the ionization state of His 55. Mutagenesis of His 55 to Lys results in a monomer in the pH range of 3 8, while the mutation to Ala results in a dimer in the same pH range. Mutations that disrupt intermolecular hydrogen bonds between Tyr 65 and Lys 44' and His 55 and Thr 67' do not change the association state of the dimer. Titration curves for His 55 and the two other histidines, His 72 and 68, were determined by (13)C (1)H NMR for H55K and for WT-LC8 in the monomeric and dimeric states. The pK(a) values of His 72 and His 68 are 6 in the WT dimer and 6.2-6.5 in monomeric H55K, while the pK(a) of His 55 is about 4.5 in the WT dimer. These results indicate that deprotonation of His 55 is linked to dimer formation and that mutation of His 55 to a small neutral residue or to a positively charged residue uncouples the protonation and dissociation processes. PMID- 16245942 TI - X-ray crystal structures of HMG-CoA synthase from Enterococcus faecalis and a complex with its second substrate/inhibitor acetoacetyl-CoA. AB - Biosynthesis of the isoprenoid precursor, isopentenyl diphosphate, is a critical function in all independently living organisms. There are two major pathways for this synthesis, the non-mevalonate pathway found in most eubacteria and the mevalonate pathway found in animal cells and a number of pathogenic bacteria. An early step in this pathway is the condensation of acetyl-CoA and acetoacetyl-CoA into HMG-CoA, catalyzed by the enzyme HMG-CoA synthase. To explore the possibility of a small molecule inhibitor of the enzyme functioning as a non-cell wall antibiotic, the structure of HMG-CoA synthase from Enterococcus faecalis (MVAS) was determined by selenomethionine MAD phasing to 2.4 A and the enzyme complexed with its second substrate, acetoacetyl-CoA, to 1.9 A. These structures show that HMG-CoA synthase from Enterococcus is a member of the family of thiolase fold enzymes and, while similar to the recently published HMG-CoA synthase structures from Staphylococcus aureus, exhibit significant differences in the structure of the C-terminal domain. The acetoacetyl-CoA binary structure demonstrates reduced coenzyme A and acetoacetate covalently bound to the active site cysteine through a thioester bond. This is consistent with the kinetics of the reaction that have shown acetoacetyl-CoA to be a potent inhibitor of the overall reaction, and provides a starting point in the search for a small molecule inhibitor. PMID- 16245943 TI - Kinetic analysis of metal binding to the amino-terminal domain of ZntA by monitoring metal-thiolate charge-transfer complexes. AB - ZntA, a P(1B)-ATPase transporter from Escherichia coli, mediates resistance specifically to Pb(2+), Zn(2+), and Cd(2+) by active efflux. ZntA has a hydrophilic N-terminal domain that binds one metal ion. This domain, approximately 120 residues long, contains the GXXCXXC motif that has been shown to be the binding site for metal ions such as Cu(+) and Zn(2+) in P(1B)-type ATPases, and an additional cysteine-rich motif, CCCDGAC. We report here that binding of Pb(2+) and Cd(2+) to this domain produces changes in the absorbance spectrum in the 250-400 nm range indicative of metal-thiolate charge-transfer complexes. The spectral changes indicate that only two cysteines are ligands to Cd(2+), but three or more cysteines are involved in binding Pb(2+); this confirms earlier results that the GXXCXXC sequence is not sufficient to bind Pb(2+), which likely involves residues from the CCCDGAC motif. The absorbance changes were used to measure metal binding kinetics of the N-terminal domain using stopped-flow techniques. Binding was described by simple second-order kinetics with a rate constant, k(on), of approximately 10(6)-10(7) M(-)(1) s(-)(1), at 4 degrees C. The activation energy of binding is similar for both Pb(2+) and Cd(2+); however, the entropy change is greater for Pb(2+). The surprisingly large rate constant for metal binding to the N-terminal domain of ZntA, compared to its low turnover rate, indicates that this step is not rate limiting in the overall transport mechanism. These results, in conjunction with earlier studies, suggest that metal binding to the transmembrane site in ZntA or metal release from the transporter is the slow step in the reaction cycle. PMID- 16245944 TI - Interaction of a peptide derived from the N-heptad repeat region of gp41 Env ectodomain with model membranes. Modulation of phospholipid phase behavior. AB - The HIV-1 gp41 envelope protein mediates the entry of the virus into the target cell by promoting membrane fusion. With a view toward possible new insights into the protein membrane alteration leading to the viral fusion mechanism, we have studied by infrared and fluorescence spectroscopies a fragment of 21 amino acids corresponding to the N-heptad repeat region of the gp41 ectodomain. Information on the structure of the peptide both in solution and in the presence of model membranes, its incorporation and location in the phospholipid bilayer, and the modulation of the phase behavior of the membrane has been gathered. Here we demonstrate that the peptide binds to and interacts with phospholipid model membranes, changing its conformation and inducing leakage of vesicle contents. These characteristics suggest that different specific regions of gp41 are capable of modifying the biophysical properties of phospholipid membranes and, therefore, might be essential for the assistance and enhancement of the viral and cell fusion process. PMID- 16245945 TI - Differential effects of temperature and hydrostatic pressure on the formation of quinonoid intermediates from L-Trp and L-Met by H463F mutant Escherichia coli tryptophan indole-lyase. AB - Escherichia coli tryptophan indole-lyase (Trpase) is a bacterial pyridoxal 5' phosphate (PLP)-dependent enzyme which catalyzes the reversible beta-elimination of l-Trp to give indole and ammonium pyruvate. H463F mutant E. coli Trpase (H463F Trpase) has very low activity with l-Trp, but it has near wild-type activity with other in vitro substrates, such as S-ethyl-l-cysteine and S-(o-nitrophenyl)-l cysteine [Phillips, R. S., Johnson, N., and Kamath, A. V. (2002) Formation in vitro of Hybrid Dimers of H463F and Y74F Mutant Escherichia coli Tryptophan Indole-lyase Rescues Activity with l-Tryptophan, Biochemistry 41, 4012-4019]. The interaction of H463F Trpase with l-Trp and l-Met, a competitive inhibitor, has been investigated by rapid-scanning stopped-flow, high-pressure, and pressure jump spectrophotometry. Both l-Trp and l-Met bind to H463F Trpase to form equilibrating mixtures of external aldimine and quinonoid intermediates, absorbing at approximately 420 and approximately 505 nm, respectively. The apparent rate constant for quinonoid intermediate formation exhibits a hyperbolic dependence on l-Trp and l-Met concentration. The rate constant for quinonoid intermediate formation from l-Trp is approximately 10-fold lower for H463F Trpase than for wild-type Trpase, but the rate constant for reaction of l-Met is similar for H463F Trpase and wild-type Trpase. The temperature dependence of the rate constants for quinonoid intermediate formation reveals that both l-Trp and l-Met have similar values of DeltaH(++), but l-Met has a more negative value of DeltaS(++). Hydrostatic pressure perturbs the spectra of the H463F l-Trp and l Met complexes, by shifting the position of the equilibria between different quinonoid and external aldimine complexes. Pressure-jump experiments show relaxations at 500 nm after rapid pressure changes of 100-400 bar with both l-Trp and l-Met. The apparent rate constants for relaxation of l-Trp, but not l-Met, show a significant increase with pressure. From these data, the value of DeltaV(++) for quinonoid intermediate formation from the external aldimine of l Trp can be estimated to be -26.5 mL/mol, a larger than expected negative value for a proton transfer. These results suggest that there may be a contribution to the deprotonation reaction either from quantum mechanical tunneling or from a mechanical coupling of protein motion and proton transfer associated with the reaction of l-Trp, but not with l-Met. PMID- 16245946 TI - Cooperative signaling among bacterial chemoreceptors. AB - Four chemoreceptors in Escherichia coli mediate responses to chemicals in the environment. The receptors self-associate and localize to the cell poles. This aggregation implies that interactions among receptors are important parameters of signal processing during chemotaxis. We examined this phenomenon using a receptor coupled in vitro assay of CheA kinase activity. The ability of homogeneous populations of the serine receptor Tsr and the aspartate receptor Tar to stimulate CheA was directly proportional to the ratio of the receptor to total protein in cell membranes up to a fraction of 50%. Membranes containing mixed populations of Tar and Tsr supported an up to 4-fold greater stimulation of CheA than expected on the basis of the contributions of the individual receptors. Peak activity was seen at a Tar:Tsr ratio of 1:4. This synergy was observed only when the two proteins were expressed simultaneously, suggesting that, under our conditions, the fundamental "cooperative receptor unit" is relatively static, even in the absence of CheA and CheW. Finally, we observed that inhibition of receptor-stimulated CheA activity by serine or aspartate required significantly higher concentrations of ligand for membranes containing mixed Tsr and Tar populations than for membranes containing only Tsr (up to 10(2)-fold more serine) or Tar (up to 10(4)-fold more aspartate). Together with recent analyses of the interactions of Tsr and Tar in vivo, our results reveal the emergent properties of mixed receptor populations and emphasize their importance in the integrated signal processing that underlies bacterial chemotaxis. PMID- 16245947 TI - 2,4,5-Trihydroxyphenylalanine quinone biogenesis in the copper amine oxidase from Hansenula polymorpha with the alternate metal nickel. AB - Copper amine oxidase (CAO) is a dual-functioning enzyme that catalyzes the biosynthesis of a self-derived coenzyme and subsequent oxidative deamination of primary amines. The organic cofactor, 2,4,5-trihydroxyphenylalanine quinone (TPQ), is generated from the post-translational modification of an active site tyrosine (Y405) in a reaction shown to be dependent on both molecular oxygen and a mononuclear copper center. Previous investigations of Cu(II)-dependent cofactor formation in the Hansenula polymorpha amine oxidase (HPAO) provided evidence for the coordination of the precursor tyrosine in forming a ligand-to-metal charge transfer complex as a means of activating the tyrosyl ring for direct attack by triplet-state dioxygen. To further delineate the role of the metal in facilitating this complex series of reactions, apo-HPAO was reconstituted with alternate metals of varying reduction potentials and Lewis acidities [Ni(II), Co(II), Mn(II), Fe(II), and Fe(III)] and the consequence of each substitution on TPQ biogenesis examined. Ni(II) was found to support the transformation of the precursor tyrosine to the quinone cofactor to yield a mature enzyme competent for methylamine oxidation. Detailed kinetic analysis of the mechanism of TPQ biogenesis for the Ni(II)-substituted enzyme has led to the proposal of a direct electron transfer from the metal-coordinated tyrosinate to dioxygen as the dominant rate-limiting step. PMID- 16245948 TI - Redox and spectroscopic properties of human indoleamine 2,3-dioxygenase and a His303Ala variant: implications for catalysis. AB - Indoleamine 2,3-dioxygenase is an important mammalian target that catalyses the oxidative cleavage of l-tryptophan to N-formylkynurenine. In this work, the redox properties of recombinant human indoleamine 2,3-dioxygenase (rhIDO) and its H303A variant have been examined for the first time and the spectroscopic and substrate binding properties of rhIDO and H303A in the presence and absence of substrate are reported. The Fe(3+)/Fe(2+) reduction potential of H303A was found to be -30 +/- 4 mV; in the presence of l-Trp, this value increases to +16 +/- 3 mV. A variety of spectroscopies indicate that ferric rhIDO at pH 6.6 exists as a mixture of six-coordinate, high-spin, water-bound heme and a low-spin species that contains a second nitrogenous ligand; parallel experiments on H303A are consistent either with His303 as the sixth ligand or with His303 linked to a conformational change that affects this transition. There is an increase in the low-spin component at alkaline pH for rhIDO, but this is not due to hydroxide bound heme. Substrate binding induces a conformational rearrangement and formation of low-spin, hydroxide-bound heme; analysis of the H303A variant indicates that His303 is not required for this conversion and is not essential for substrate binding. The Fe(3+)/Fe(2+) reduction potential of H303A variant is approximately 70 mV lower than that of rhIDO, leading to a destabilization of the ferrous-oxy complex, which is an obligate intermediate in the catalytic process. In comparison with the properties of other heme enzymes, the data can be used to build a more detailed picture of substrate binding and catalysis in indoleamine 2,3-dioxygenase. The wider implications of these results are discussed in the context of our current understanding of the catalytic mechanism of the enzyme. PMID- 16245949 TI - Relative affinities of DNA sequences for the histone octamer depend strongly upon both the temperature and octamer concentration. AB - Using a novel competition assay to determine the relative strength of different histone octamer-binding sites, we have compared three natural and two synthetic sites. We show that the relative affinities of these sites for the histone octamer depend upon both the temperature and octamer concentration. In particular, under certain conditions, a natural octamer-binding site from a yeast promoter outcompetes a synthetic sequence of comparable affinity to the strongest previously described positioning sequence. Under other conditions, this synthetic sequence is the preferred octamer ligand. We infer that sequence selection by the histone octamer depends strongly upon both the sequence-dependent anisotropy of DNA bending and on DNA deformability and that these parameters may contribute differently to nucleosome formation. These findings indicate that previous studies designed to identify strong octamer-binding sites may fail to select some natural strong binding sites. PMID- 16245950 TI - DNA damage levels and biochemical repair capacities associated with XRCC1 deficiency. AB - Base excision repair (BER) is the major corrective pathway for most spontaneous, oxidative, and alkylation DNA base and sugar damage. X-ray cross-complementing 1 (XRCC1) has been suggested to function at nearly every step of this repair process, primarily through direct protein-protein interactions. Using whole cell extract (WCE) repair assays and DNA damage measurement techniques, we examined systematically the quantitative contribution of XRCC1 to specific biochemical steps of BER and single-strand break repair (SSBR). Our studies reveal that XRCC1 deficient Chinese hamster ovary WCEs exhibit normal base excision activity for 8 oxoguanine (8-OH-dG), 5-hydroxycytosine, ethenoadenine, and uracil lesions. Moreover, XRCC1 mutant EM9 cells possess steady-state levels of endogenous 8-OH dG base damage similar to those of their wild-type counterparts. Abasic site incision activity was found to be normal in XRCC1-deficient cell extracts, as were the levels of abasic sites in isolated chromosomal DNA from mutant cells. While one- and five-nucleotide gap filling was not affected by XRCC1 status, a significant approximately 2-4-fold reduction in nick ligation activity was observed in EM9 WCEs. Our results herein suggest that the primary biochemical defect associated with XRCC1 deficiency is in the ligation step of BER/SSBR, and that XRCC1 plays no significant role in endogenous base damage and abasic site repair, or in promoting the polymerase gap-filling step. PMID- 16245951 TI - Unfolding and refolding in vitro of a tetrameric, alpha-helical membrane protein: the prokaryotic potassium channel KcsA. AB - 2,2,2-Trifluoroethanol (TFE) effectively destabilizes the otherwise highly stable tetrameric structure of the potassium channel KcsA, a predominantly alpha-helical membrane protein [Valiyaveetil, F. I., Zhou, Y., and MacKinnon, R. (2002) Biochemistry 41, 10771-10777]. Here, we report that the effects on the protein structure of increasing concentrations of TFE in detergent solution include two successive protein concentration-dependent, cooperative transitions. In the first of such transitions, occurring at lower TFE concentrations, the tetrameric KcsA simultaneously increases the exposure of tryptophan residues to the solvent, partly loses its secondary structure, and dissociates into its constituent subunits. Under these conditions, simple dilution of the TFE permits a highly efficient refolding and tetramerization of the protein in the detergent solution. Moreover, following reconstitution into asolectin giant liposomes, the refolded protein exhibits nativelike potassium channel activity, as assessed by patch clamp methods. Conversely, the second cooperative transition occurring at higher TFE concentrations results in the irreversible denaturation of the protein. These results are interpreted in terms of a protein and TFE concentration-dependent reversible equilibrium between the folded tetrameric protein and partly unfolded monomeric subunits, in which folding and oligomerization (or unfolding and dissociation in the other direction of the equilibrium process) are seemingly coupled processes. At higher TFE concentrations this is followed by the irreversible conversion of the unfolded monomers into a denatured protein form. PMID- 16245952 TI - Point mutations in apolipoprotein A-I mimic the phenotype observed in patients with classical lecithin:cholesterol acyltransferase deficiency. AB - We have analyzed the effect of charged to neutral amino acid substitutions around the kinks flanking helices 4 and 6 of apoA-I and of the deletion of helix 6 on the in vivo activity of LCAT and the biogenesis of HDL. The LCAT activation capacity of apoA-I in vitro was nearly abolished by the helix 6 point (helix 6P apoA-I[R160V/H162A]) and deletion {helix 6Delta-apoA-I[Delta(144-165)]} mutants, but was reduced to 50% in the helix 4 point mutant (helix 4P-apoA I[D102A/D103A]). Following adenovirus-mediated gene transfer in apoA-I deficient mice, the level of plasma HDL cholesterol was greatly reduced in helix 6P and helix 6Delta mutants. Electron microscopy and two-dimensional gel electrophoresis showed that the helix 6P mutant formed predominantly high levels of apoA-I containing discoidal particles and had an increased prebeta1-HDL/alpha-HDL ratio. The helix 6Delta mutant formed few spherical particles and had an increased prebeta1-HDL/alpha-HDL ratio. Mice infected with adenovirus expressing the helix 4P mutant or wild-type apoA-I had normal HDL cholesterol and formed spherical alpha-HDL particles. Coinfection of mice with adenoviruses expressing human LCAT and the helix 6P mutant dramatically increased plasma HDL and apoA-I levels and converted the discoidal into spherical HDL, indicating that the LCAT activity was rate-limiting for the biogenesis of HDL. The LCAT treatment caused only a small increase in HDL cholesterol and apoA-I levels and in alpha-HDL particle numbers in the helix 6Delta mutant. The findings indicate a critical contribution of residue 160 of apoA-I to the in vivo activity of LCAT and the subsequent maturation of HDL and explain the low HDL levels in heterozygous subjects carrying this mutation. PMID- 16245953 TI - Structural determinants of cross-linking and hydrophobic domains for self assembly of elastin-like polypeptides. AB - Elastin is a major structural protein found in large blood vessels, lung, ligaments, and skin, imparting the physical properties of extensibility and elastic recoil to these tissues. To achieve the required structural durability of the elastic matrix, the elastin monomer, tropoelastin, undergoes ordered assembly into a covalently cross-linked, fibrillar polymeric structure. Human tropoelastin consists of 34 exons coding for alternating hydrophobic and cross-linking domains. Using a series of well-defined recombinant polypeptides based on human elastin sequences mimicking native elastin, we have previously investigated the role of sequence and context of hydrophobic domains in elastin self-assembly. Here, we demonstrate that the structure of both cross-linking and hydrophobic domains have significant effects on the assembly of these polypeptides. Removing a putative flexible hinge region in the center of a cross-linking domain substantially increased the alpha-helical content and strongly promoted their self-aggregation. However, while trifluoroethanol (TFE) promoted and urea inhibited self-assembly of these polypeptides, these effects were not predominantly due to altered alpha-helicity of the polypeptides. Our results suggest that, while increased alpha helicity also favors this process, the major effect of TFE to promote organized self-assembly of elastin-like polypeptides is likely related to direct effects of this cosolvent on hydrophobic domains. Such simple elastin polypeptide models can provide an important tool for understanding the relationships between sequence, structure, and polymeric assembly of elastin. PMID- 16245954 TI - Role of oxysterol structure on the microdomain-induced microsolubilization of phospholipid membranes by apolipoprotein A-I. AB - Oxygenated derivatives of cholesterol, oxysterols, have different physicochemical properties and three-dimensional shapes. The kinetics of microsolubilization of dimyristoylphosphatidylcholine (DMPC) multilamellar vesicles by apolipoprotein A I (apoA-I) to form discoidal high-density lipoproteins (rHDL) was dramatically affected by oxysterol chemical structure. Under the experimental conditions of varying oxysterol chemical structure, sterol concentration, and the lipid phase state of DMPC, the kinetics varied over 3 orders of magnitude. Some oxysterols behaved similarly to cholesterol and increased the rate of microsolubilization; however, they were not as effective as cholesterol. Other oxysterols greatly inhibited this process. In general, there was no correlation of the rates with membrane fluidity as measured by fluorescence polarization. The rate of DMPC microsolubilization by apoA-I is highly dependent upon the presence of lattice defects in the membrane surface that occur due to imperfect packing of coexisting lipid phases. The differential ability of various oxysterols to induce the formation of an ordered lipid phase is the probable basis for their effects on the rates of DMPC microsolubilization. There was no effect of oxysterol chemical structure on the structure of the equilibrium rHDL products; however, there was a dramatic effect of sterol concentration on rHDL particle size. Different oxysterols regulate the kinetics of apoA-I membrane association by altering structural microheterogeneity at the membrane surface. However, once the kinetic barrier is overcome, the particle sizes of rHDL products formed are determined solely by the amount of sterol presence. PMID- 16245955 TI - Asymptomatic bacteriuria in elderly patients: significance and implications for treatment. AB - Asymptomatic bacteriuria (ASB) is frequent in elderly patients and even more prevalent in residents of long-term care facilities. Furthermore, because more and more people are reaching advanced age and the need for care increases with age, ASB is becoming increasingly important. There are several definitions for ASB, all of which require positive urine cultures and place little or no importance on accompanying pyuria. Most ASB is associated with complicating factors, as might be found in complicated urinary tract infections (UTIs). Thus, the bacterial spectrum associated with ASB is comparable to that seen in complicated UTIs. A variety of complicating factors are more frequently found in elderly patients with ASB, such as hormonal factors (e.g. estrogen decrease), certain anatomical factors (e.g. prostate obstruction), metabolic factors (e.g. diabetes mellitus), functional alteration of the urinary bladder, immunological changes and a high rate of indwelling-catheter use. Screening for ASB in elderly people is limited to those undergoing invasive urological procedures and surgical procedures with implant material. In other situations, examination of the urine is not recommended if signs or symptoms in the urinary tract are absent. Treatment of ASB is recommended only before urological procedures. Pyuria accompanying ASB is not an indication for antimicrobial treatment. If antimicrobial treatment is considered, concomitant factors that occur frequently in elderly people, such as renal insufficiency, must be taken into account. Although ASB is apparently a benign condition, prevention in elderly people is important. The degree of pathogenicity of bacteria causing ASB has not yet been satisfactorily elucidated. Therefore, until the implications of the bacteria involved in ASB are fully understood, implementing the same hygienic precautions as are used in individuals with symptomatic UTIs should at least be undertaken. PMID- 16245956 TI - Management issues with elderly-onset rheumatoid arthritis: an update. AB - Elderly-onset rheumatoid arthritis (EORA) is defined as rheumatoid arthritis (RA) starting at >60 years of age. EORA is characterised by a lower female/male ratio compared with RA in younger patients and it more frequently has an acute onset accompanied by constitutional symptoms. Two incompletely overlapping subsets of RA have been recognised: one exhibits the classical RA clinical picture while the other has a polymyalgia rheumatica-like appearance, characterised by shoulder involvement, absence of rheumatoid factor and, usually, by a nonerosive course. Identification of anti-cyclic citrullinated peptide antibodies is useful for distinguishing the latter subset from true polymyalgia rheumatica. Elderly-onset spondyloarthritis, crystal-related arthritis, remitting seronegative symmetrical synovitis with pitting oedema syndrome and hepatitis C virus-related arthritis must also be considered in the differential diagnosis. EORA treatment requires prudence because of the increase in age-related risks pertaining principally to the renal, cardiovascular and gastrointestinal systems. No groups of molecules usually employed for RA therapy in younger subjects (analgesics, NSAIDs, corticosteroids, disease-modifying antirheumatic drugs, anticytokine drugs) can be excluded a priori from the treatment of EORA patients. Nevertheless, the risk/benefit ratio relating to their use must be accurately evaluated for every single patient. Recently marketed compounds such as leflunomide and tumour necrosis factor-alpha antagonists have also increased the therapeutic opportunities for aged RA patients. PMID- 16245958 TI - Antipsychotics for behavioural and psychological problems in elderly people with dementia: a systematic review of adverse events. AB - OBJECTIVE: Although antipsychotics are important in the treatment of behavioural and psychological symptoms of dementia (BPSD), they have moderate efficacy and often cause adverse events. Recent safety warnings about increased frequency of cerebrovascular adverse events in elderly patients who use atypical antipsychotics mean that physicians now face a dilemma when weighing the benefits and risks of use of antipsychotics in this patient group. This study systematically reviews the reporting of adverse events of antipsychotics used to treat BPSD in randomised, controlled trials (RCTs). METHODS: We searched the MEDLINE, EMBASE, PsychInfo and CINAHL databases (search period 1980 or 1986-April 2005) and the Cochrane controlled trials register (2005) for RCTs that used intention-to-treat analysis to evaluate the efficacy and harms of antipsychotics used to treat BPSD. Two independent reviewers assessed the reporting of adverse events. RESULTS: Screening of 930 abstracts identified 12 eligible RCTs (2809 patients). Most participants were elderly people (mean age 80 years) with Alzheimer's, vascular or mixed dementia. Studies lasted from 3 to 16 weeks. Adverse events, though common, were described heterogeneously and incompletely. No RCT fulfilled all Consolidated Standards of Reporting Trials requirements for the reporting of harms. Atypical antipsychotics caused fewer extrapyramidal symptoms and less somnolence than typical antipsychotics, but these differences disappeared when dosages were increased. Only one trial reported cerebrovascular adverse events, with a number needed to harm of 14 (95% CI 8, 41). CONCLUSIONS: At lower doses atypical antipsychotics may cause fewer adverse events in the treatment of BPSD, but there is uncertainty about their cerebrovascular safety profile. The RCTs included in this systematic review described adverse events too incompletely and heterogeneously to allow generation of clear treatment recommendations, and they do not provide sufficient evidence to support recent safety warnings. Better reporting on harms in RCTs is needed to enable rational treatment decisions with respect to use of antipsychotics for BPSD. PMID- 16245957 TI - Aetiology and management of male erectile dysfunction and female sexual dysfunction in patients with cardiovascular disease. AB - The historical basis for understanding erectile function as a neurovascular phenomenon and the advance from fanciful to effective treatment of erectile dysfunction (ED) are reviewed, with emphasis on patients with cardiovascular disease (CVD). ED occurs in 60% of CVD patients by 40 years of age. Male ED and female sexual dysfunction (FSD) diminish quality of life and often warn of occult CVD. ED is often unrecognised but is readily diagnosed during a 5-minute interview using a truncated International Index of Erectile Function questionnaire. Erection of the penis and clitoral engorgement result from local, arousal-induced release of neuronal and endothelial-derived nitric oxide (NO). Arterial vasodilatation and relaxation of cavernosal smooth muscle cells cause arterial blood to flood trabecular spaces, compressing venous drainage, resulting in tumescence. Cyclic guanosine monophosphate (cGMP)-induced activation of protein kinase G mediates the effects of NO by enhancing calcium sequestration and activating large-conductance, calcium-sensitive K+ channels. Future treatment strategies will likely enhance these pathways. Phosphodiesterase-5 inhibitors (sildenafil, tadalafil and vardenafil) increase cGMP levels in erectile tissue. These agents are effective in 80% of CVD patients with ED and can be used safely, even in the presence of stable coronary disease or congestive heart failure, provided nitrates are avoided and patients do not have hypotension, severe aortic stenosis or evocable myocardial ischaemia. Second-line therapies (vacuum constrictor device and transurethral or intracavernosal prostaglandin E1) can also be used in CVD patients. Treatment of FSD and its relationship to CVD are less well established, but similarities to ED exist. ED can be prevented by reduction of CVD risk factors, exercise, weight loss and abstinence from smoking. PMID- 16245960 TI - Outpatient medications and hip fractures in the US: a national veterans study. AB - BACKGROUND: Certain medications have been linked to falls. One of the most severe fall-related injuries in the elderly is a hip fracture. OBJECTIVE: The objective of this study was to examine the use of medications known to increase fall risk that were prescribed on an outpatient basis to veterans prior to hospital admission for hip fracture. METHODS: We identified and analysed the use of outpatient fall-related medications in 2212 unique patients with hip fractures admitted in fiscal year 2003 to Veterans Health Administration hospitals compared with that for matched controls (exact match for age and sex) admitted for acute myocardial infarction (MI) or pneumonia. We analysed the medications selected from the three drug categories most often linked with an increased risk for falls. These categories included medications that affected the cardiovascular (CVS), CNS or the musculoskeletal system (MSS). The unit of analysis was the hip fracture linked with outpatient medications in the study group compared with matched control groups of patients with hospitalisations for an acute MI or pneumonia. RESULTS: Of the 2212 hip-fracture patients, 70% had fall-related medications prior to hospitalisation for hip fracture. The most notable differences in usage were seen in the drug classes antiepileptics/barbiturates, antidepressants (2-fold difference in use of selective serotonin reuptake inhibitors [SSRIs] and tricyclic antidepressants [TCAs]), and antiparkinson's drugs (nearly 4-fold difference between cases and controls). There were also notable differences in usage of antipsychotics (3-fold difference for hip fracture cases compared with acute MI) and cholinesterase inhibitors (nearly 2 fold difference for hip-fracture cases compared with pneumonia or acute MI). The most notable differences in polypharmacy combinations were CVS and CNS categories with differences of 9.44% (absolute) and 43% (relative) for hip-fracture patients over acute MI, and 4.83% (absolute) and 18% (relative) for hip-fracture patients over pneumonia patients. CONCLUSIONS: This is the first national Veterans Health Administration hip fracture hospitalisation study that temporally linked outpatient fall-related medications in hip-fracture patients with matched controls. We found that of veterans with hip-fracture hospitalisations, 70% were prescribed outpatient medications from selected major drug categories that may potentially increase fall risk. Moreover, over one-third of hip-fracture patients received concomitant prescriptions of drugs from multiple selected drug categories. Hip-fracture patients, compared with matched controls of acute MI and pneumonia, had the largest pronounced differences in prescribed medications in the antiepileptics, antidepressants, antipsychotics and antiparkinson's drug classes. Although a randomised clinical trial is the 'gold standard' for determining causation issues, exposing patients, particularly the elderly, to the potential risk of injurious falls would raise serious patient safety research approval issues. If the relationship between selected drugs and falls is indeed to some extent causative, future retrospective multivariate analyses could quantify the magnitude of these effects. PMID- 16245962 TI - An ex-vivo, whole-globe porcine model of corneoepithelial wound healing tested using immunomodulatory drugs. AB - BACKGROUND: An efficient epithelial wound healing is essential for the preservation of vision. Hence, the effects of novel topical drugs on the ocular surface must be ascertained before clinical use. We have tested the utility of an ex vivo, whole-globe porcine screening model to serve as a partial substitute for resource- and time-consuming animal experiments. METHODS: Standardized corneoepithelial lesions, 5.0 mm in diameter and 40 microm in depth, were created with an Excimer laser in freshly enucleated porcine eyes. These were then exposed to control solutions (physiological saline (baseline), tissue-culture medium (positive control) and NH4 + (toxicity control)) and to three test agents (cyclosporin A, dexamethasone, and mitomycin C). The wound-healing response and toxic effects were monitored after 20-26 h by comparing lesion sizes. RESULTS: According to baseline data obtained using physiological saline, tissue-culture medium improved wound healing. The highest doses of NH4 + (1 M) and mitomycin C (1.0 mg/mL) elicited toxic effects (confidence interval according to Scheffe's post hoc test: -0.65 to -0.07 and -0.99 to -0.60, respectively). Under the same test conditions, cyclosporin A (0.1 to 10 mg/mL) and dexamethasone (0.1 to 10 mg/mL) had no influence on corneoepithelial wound healing. CONCLUSIONS: Drug screening with this ex vivo porcine model permits a reproducibly quantitative and time- and dose-dependent assessment of corneoepithelial wound healing. This model corresponds more closely to the clinical situation than cell culturing and may, therefore, be useful in evaluating novel pharmaceutical agents, thereby helping to cut down on the number of animal experiments performed prior to the instigation of clinical trials. PMID- 16245961 TI - Effects of SV40 T antigen transduction on human corneal endothelial cell wound healing in vitro. AB - OBJECTIVE: The aim of this study was to investigate the endothelial wound healing effects of SV40 large T and small t antigen transduction on cultured human corneal endothelial cells (HCEC). METHODS: Human corneal endothelial cells were infected with either mock solution, Ad green fluorescent protein (GFP), or Ad SV40 T/t antigen/GFP, then mechanically wounded 48 h later. The endothelial wound healing rate was quantified by an analysis of the photographs taken every 12 h after wounding. The characteristics of Ad SV40 T/t Ag/GFP-infected human corneal endothelial cells were evaluated with cell morphology, cell density, contact inhibition, and cytoskeletal features using rhodamine phalloidin to stain F actin. DNA synthesis was assessed using 5-Bromo-2'-deoxy-uridine (BrdU) labeling. RESULTS: Wound healing rates in the first 12 and 24 h after wounding were significantly faster in the Ad SV40 T/t antigen/GFP-infected group than the other two groups. In all three groups, the morphology, cell density, and cytoskeletal features of cells at confluency was similar and contact inhibition retained. There were no differences in the pattern of F-actin and endothelial cell density 4 d after wound closure. However, during the process of wound healing, prominent stress fibers in migrating cells near the wound edge were noted in normal cells at 36 h after wounding, whereas the Ad SV40 T/t Ag/GFP-infected cells showed similar changes as early as 12 h after wounding. BrdU staining results revealed that the Ad SV40 T/t antigen/GFP-infected group had labeled cells showing DNA synthesis in the wound area at 12 h after wounding, while no labeled cells were found in the other two groups. CONCLUSIONS: In an in vitro model, transduction of human corneal endothelial cells using a recombinant adenoviral vector expressing SV40 T/t antigen enhanced both the wound healing rate and proliferative capacity, especially in the first 12 h after wounding, and the characteristic morphologic features of the infected cells were maintained. PMID- 16245963 TI - Effects of various lubricants on corneal surface regularity in rabbits. AB - The aim of this study was to establish a method for analysis of tear film stability and thereby to investigate the effects of lubricants on corneal surface regularity and corneal epithelial damage in anesthetized rabbits with the eye held open. Saline-instilled and noninstilled eyes manifested a time-dependent increase in the corneal surface regularity index (SRI). In contrast, 0.1% hyaluronic acid (HA) prevented the increase in the SRI for up to 20 min after instillation. Methylene blue uptake into the damaged cornea was significantly greater in eyes that received saline than in those treated with HA. Whereas eyes instilled with 0.5% methylcellulose (MC) or 0.35% sodium carboxymethylcellulose (CMC) manifested an increase in the SRI similar to that apparent in those that received saline, 1.0% chondroitin sulfate (CS) prevented the increase in the SRI for up to 10 min after instillation. In conclusion, we have established a method for analysis of tear film stability in rabbits. Our results suggest that tear film stability is closely related to corneal surface damage and that topical instillation of 0.1% HA or 1.0% chondroitin sulfate warrants further investigation as a potential treatment for patients with dry eye. PMID- 16245959 TI - Prevention of cardiovascular events in elderly people. AB - BACKGROUND AND OBJECTIVE: Cardiovascular disease has been identified as the leading cause of morbidity and mortality in developed countries. Given the increase in life expectancy and the development of cardiovascular preventive measures, it has become increasingly important to detect and prevent cardiovascular diseases in the elderly. We reviewed the scientific literature concerning cardiovascular prevention to assess the importance of cardiovascular preventive measures in old (> or =65 years of age) individuals. METHODS: We undertook a systematic search for references relating to prevention of cardiovascular disease in the elderly, mainly ischaemic stroke, coronary artery disease and heart failure, on the MEDLINE database 1962-2005. For cardiovascular prevention by drugs or surgery, emphasis was placed on randomised controlled trials, review articles and meta-analyses. For cardiovascular prevention by lifestyle modification, major cohort studies were also considered. RESULTS: Stroke, coronary heart disease and heart failure were found to be the main targets for cardiovascular prevention in published studies. Antihypertensive treatment has proven its efficacy in primary prevention of fatal or nonfatal stroke in hypertensive and high-risk patients >60 years of age, particularly through treatment of systolic hypertension. Systolic blood pressure reduction is equally important in the secondary prevention of stroke. Similarly, in nonvalvular atrial fibrillation, an adjusted dose of warfarin with a target International Normalized Ratio (INR) of between 2 to 3 prevents ischaemic stroke in elderly patients with an acceptable haemorrhagic risk but is still under prescribed. Antiplatelet agents are indicated in elderly patients with nonembolic strokes. Few large-scale studies have investigated the effect of HMG-CoA reductase inhibitors (statins) on stroke prevention in old individuals. To date, the largest trials suggest a beneficial effect for stroke prevention with use of statins in high-risk elderly subjects < or =82 years of age. Carotid endarterectomy is indicated in carotid artery stenosis >70% and outcomes are even better in elderly than in younger patients. However, medical treatment is still the first-line treatment in asymptomatic elderly patients with <70% stenosis. In ischaemic heart disease, different trials in elderly individuals have shown that use of statins, antithrombotic agents, beta-adrenoceptor antagonists and ACE inhibitors plays an important role either in primary or in secondary cardiovascular prevention. Hormone replacement therapy has been used to treat climacteric symptoms and postmenopausal osteoporosis and was thought to confer a cardiovascular protection. However, controlled trials in elderly individuals changed this false belief when it was found that there was no benefit and even a harmful cardiovascular effect during the first year of treatment. Smoking cessation, regular physical activity and healthy diet are, as in younger individuals, appropriate and effective measures for preventing cardiovascular events in the elderly. Finally, antihypertensive treatment and influenza vaccination are useful for heart failure prevention in elderly individuals. CONCLUSIONS: Cardiovascular prevention should be more widely implemented in the elderly, including individuals aged > or =75 years, and this might contribute to improved healthy status and quality of life in this growing population. PMID- 16245964 TI - Leptin in corneas from keratoconus and infectious keratitis patients. AB - PURPOSE: Leptin is produced primarily by adipose tissue. More recent studies have shown extra sites of leptin production in physiologic and ill human tissues. However, whether leptin originates from human corneas in infectious keratitis and keratoconus is not known. The aim of this study was to demonstrate and quantitate leptin expression in corneas with infectious keratitis and keratoconus and make comparisons to control corneas. METHODS: We examined the immunohistochemical staining of leptin in nine corneas surgically excised from patients with infectious keratitis (3 patients), keratoconus (3 patients), and donor corneas (3 patients). RESULTS: The results were analyzed using a semiquantitative scoring system of mild, moderate, and strong. Cells of the infectious keratitis group had the strongest leptin staining intensity, the control group had moderate, and the keratoconus group had mild staining intensity. The more vascular corneas in the infectious keratitis group were also associated with the greatest leptin staining. CONCLUSIONS: Our findings indicate that leptin expression was present in all three sources of corneas (infectious keratitis, keratoconus, and normal control). Quantitative scoring would imply it may play a role in infectious keratitis, although further experiments are necessary to establish any causal relationship. PMID- 16245965 TI - The efficacy of timolol in gel-forming solution after morning or evening dosing in Asian glaucomatous patients. AB - AIMS: The aim of this study was to determine the effectiveness of timolol gel forming solution as a monotherapy and to compare the efficacy of timolol gel forming solution morning instillation versus at night application in Asians with newly diagnosed open-angle glaucoma. METHODS: A retrospective, comparative, parallel group study was conducted on 78 eligible patients diagnosed with open angle glaucoma who received topical timolol gel-forming solution either on morning (AM dosing group; 39 patients) or evening (PM dosing group; 39 patients) as monotherapy treatment. Intraocular pressure was taken at baseline, 1, 3, 6, and 12 months post-treatment. RESULTS: Mean baseline intraocular pressure (IOP) for timolol gel-forming solution in general was 22.7 +/- 6.6 mmHg with the mean IOP reduction of 5.7 +/- 5.3 mmHg (23.1% reduction). The mean IOP reduction of the AM dosing group was 7.5 +/- 5.2 mmHg (31.0% reduction) and 3.9 +/- 4.9 mmHg (15.1% reduction) in the PM dosing group (P = 0.0002). There was a statistically significant difference of IOP reductions between AM and PM dosing group at 1 month (P = 0.041), 3 months (P = 0.037) and 12 months (P = 0.015) post-treatment. Seventy-five percent (24) of good responders was in the AM group and 67.4% (31) of poor responders were patients in the PM group. CONCLUSION: Timolol gel-forming solution is effective in reducing IOP in Asian eyes. Morning dosing was more effective in lowering the IOP compared to evening dosing. PMID- 16245966 TI - The efficacy and ocular discomfort of substituting brinzolamide for dorzolamide in combination therapy with latanoprost, timolol, and dorzolamide. AB - PURPOSE: The aim of this study was evaluate the efficacy and ocular discomfort of substituting brinzolamide for dorzolamide in patients with glaucoma treated by latanoprost, timolol, and dorzolamide. METHODS: An 8-week, prospective, randomized, open-label, comparative study was performed in 58 patients with primary open-angle glaucoma treated by latanoprost, timolol, and dorzolamide. These patients were randomly enrolled into two groups: (1) dorzolamide three times daily was substituted with brinzolamide twice-daily (substituting group); and (2) dorzolamide three times daily was continued (control group). Intraocular pressure (IOP) was measured at baseline, 4, and 8 weeks after the enrollment. Subjective ocular discomfort (irritation and blurred vision) at the time of the instillation of the patient was noted with interview. RESULTS: The IOPs at baseline, 4 and 8 weeks after the enrollment were 17.7 +/- 2.7 mmHg, 17.5 +/- 2.6 mmHg, and 17.4 +/- 2.9 mmHg in the substituting group, and 18.0 +/- 2.5 mmHg, 17.8 +/- 2.5 mmHg, and 17.9 +/- 2.6 mmHg in the control group, respectively. There were no significant differences in IOP changes between the two groups (P = 0.74). In the substituting group, ocular irritation was decreased significantly (P = 0.0014) from 63% to 20%. The slight increase of blurred vision from 27% to 37% that occurred in the substituting group was not significant (P = 0.58). In the control group, neither ocular irritation (P = 0.58, from 68% to 57%) nor blurred vision (P = 0.99, from 25% to 21%) was changed. CONCLUSIONS: Substituting brinzolamide for dorzolamide maintained stable IOP with improvement in ocular comfort in patients with glaucoma. PMID- 16245967 TI - Efficiency of olopatadine hydrochloride 0.1% in the treatment of vernal keratoconjunctivitis and goblet cell density. AB - PURPOSE: The aim of this study was to investigate the effect of topical 0.1% olopatadine hydrochloride on goblet cell density, clinical signs, and symptoms of patients with vernal keratoconjunctivitis. METHODS: Between December 2002 and April 2003, 40 eyes of 20 patients with vernal keratoconjunctivitis and 10 healthy eyes of 5 control patients were evaluated prospectively and treated with 0.1% olopatadine hydrochloride. Both groups were observed clinically, subjective complaints were recorded, and changes in goblet cell density were obtained with brush cytology. RESULTS: After the 2-month therapy, subjective complaints and clinical signs improved with therapy. Also, the clinical signs were improved with the therapy. As the severity of the signs and symptoms were reduced, goblet cell numbers in the brush cytologic specimens were reduced. CONCLUSION: Olopatadine hydrochloride 0.1% is an effective agent for relieving the signs and symptoms of vernal keratoconjunctivitis. Also, it reduces the number of goblet cells, which, in turn, decreases the amount of mucus discharge in vernal keratoconjunctivitis during treatment. PMID- 16245968 TI - Evaluation of mitomycin C in reducing postoperative adhesions in strabismus surgery. AB - AIMS: The aim of this study was to evaluate the efficacy of mitomycin C in reducing postoperative adhesions following strabismus surgery. METHODS: This study was a prospective, single-center, interventional case series. We randomized 14 patients with restrictive strabismus to the mitomycin C group (7 patients) or to the control group (7 patients). We used mitomycin C intraoperatively for 5 min at a concentration of 0.2 mg/mL in the mitomycin group after adhesion release. We compared postoperative results using the forced duction test. RESULTS: Compared to the control group, the mitomycin C group exhibited a better range of passive duction at all postoperative follow-up times; however, mitomycin C did not completely eliminate the problem of postoperative adhesions. Patients receiving mitomycin C did not report any complications during this study. CONCLUSIONS: The intraoperative application of mitomycin C is a safe and effective adjunct to surgery in the treatment of restrictive strabismus. PMID- 16245970 TI - Concerns, satisfaction, and retention of Canadian community health nurses. AB - This study of Canadian community health nurses (N = 1,044) compared the work related concerns, job satisfaction, and factors influencing the retention of public health, home care, and community care access center (CCAC) nurses. Community health nurses identified similar work-related issues as being of greatest concern to them, but there were significant differences among the 3 groups of nurses in the magnitude of these concerns. There were also significant differences among the 3 groups for satisfaction with their jobs and their immediate supervisors, with CCAC nurses being the least satisfied except for the greater dissatisfaction of home care nurses with their pay and benefits. For the retention factors, the differences were primarily in the areas of job features and supportive work relationships. There are both similarities and differences among public health, home care, and CCAC nurses. Initiatives to address community health nurses' concerns, improve their job satisfaction, and increase their retention will require interventions tailored to the specific community health care setting. PMID- 16245971 TI - Smoking initiation prevention among youths: implications for community health nursing practice. AB - Cigarette smoking among youths has long been documented as a national problem affecting health and economic status in the United States. A number of studies have documented that cigarette-smoking initiation occurs primarily between late childhood and young adolescence. This evidence has brought about the need for awareness among community health nurses to find and deliver effective antismoking programs to reduce the prevalence of youth smoking initiation. Generally, community health nurses are in an excellent position to help the nation achieve its goals in terms of reducing the incidence of youth smoking initiation. However, current knowledge about community health nursing practice and smoking initiation interventions is limited. This article raises awareness about smoking initiation prevention in youth and the need to implement effective smoking prevention programs in practice settings and encourages community health nurses to increase their involvement in antismoking initiation research and interventions. PMID- 16245972 TI - School nurses' opinions about the prevention of tobacco use. AB - To further understand school nurses' tobacco policy beliefs and attitudes toward tobacco companies, a convenience sample of 53 school nurses completed questionnaires about anti-tobacco policies and attitudes toward tobacco companies. Overall, these nurses strongly agreed with tobacco policies such as banning youths from wearing clothing with a tobacco logo to school and fining restaurant owners who permit smoking. In addition, these nurses on average were negative toward tobacco companies. For example, they thought that tobacco companies and advertising leads to youth tobacco use. However, this group of school nurses thought that youths were basically neutral toward tobacco companies. Considering their professional experience with tobacco prevention at school, school nurses would be logical designers of tobacco prevention school interventions. An implication of these results is that school nurse education should include policy and activism components. PMID- 16245974 TI - A study of the level of knowledge about diabetes management of low-income persons with diabetes. AB - Diabetes education has proven to be an effective intervention for the management of diabetes. Persons with lower education levels, the majority of whom have low incomes, receive less diabetes self-management education (DSME) than their peers. This study compared the diabetes knowledge level of low-income adults who did not have a high school diploma or general equivalency diploma with those who did. We conducted a survey among residents of 4 low-income neighborhoods in Cincinnati, Ohio, using a cross-sectional descriptive design. Surveys included information related to diabetes self-management and level of education. The only questions for which there was a significant difference between the 2 groups were those pertaining to the causes and treatment of hypoglycemia. Survey results were used to validate the need for a DSME program for low-income adults in Cincinnati, Ohio. A DSME program was developed specifically for persons with low literacy. PMID- 16245973 TI - Mental health for older adults and benzodiazpine use. AB - Benzodiazepine (BZD) drug use among community-dwelling seniors is a significant health issue. Although long-term use of BZDs by seniors is a recognized problem, little is known about the mental health of the consumers. Better knowledge of their mental health would help nurses in identifying the psychological needs of this population. The goals of this longitudinal study1 (n = 138) were to describe the mental health status of long-term users of BZDs and to compare it with the mental health of seniors who have either begun or stopped consuming BZDs over a 1 year period (from Phase 1 to Phase 2). Results showed that one third of long-term users of BZDs do not present any mental health problem. Furthermore, no differences were observed between the mental health statuses of new users of BZDs, individuals who stopped using BZDs, and long-term users of BZDs. In conclusion, at least one third of long-term users of BZDs should stop using these drugs, and nurses should play a leading role in helping these seniors withdraw from BZD consumption. PMID- 16245975 TI - Community leaders' perceptions of single, low-income mothers' needs and concerns for social support. AB - Purposes of this study were to (a) assess and describe needs and concerns of single, low-income mothers during their transition to parenthood from the perspective of 16 Midwestern community leaders working closely with families and (b) evaluate social support mechanisms that are available for families. Focus group questions were organized around social support theory to gather information. The following themes evolved from focus group discussions: (a) social support issues (emotional, tangible, informational, and appraisal support; positive and negative support); (b) personal barriers to success (stress, low self-esteem, isolation, and inadequate parenting competence); and (c) system barriers (fear of the system and insensitive and ineffective services). Community health nursing strategies were identified that include assessment and interventions for this vulnerable population throughout their infants' 1st year of life and beyond. PMID- 16245986 TI - Attention deficit and hyperactivity disorder among school children in Kinshasa, Democratic Republic of Congo. AB - OBJECTIVES: To estimate the prevalence and determinants of attention deficit and hyperactivity disorder (ADHD) symptoms among school children in Kinshasa, an African urban setting. METHODS: The 18-items of the Disruptive Behaviour Disorder rating scale (DBD), which is based on the Diagnostic and Statistical Manual for mental disorders 4(th) edition (DSM-IV), were used to investigate the presence of ADHD symptoms. Parents interviews, using a questionnaire specially designed for the study, were performed to identify socio-demographic characteristics. All children were subject to a clinical examination. RESULTS: The estimated prevalence of DSM-IV ADHD symptoms was 6%. Those with family health problems, younger age at start of primary school, good nutritional status and poor school performance more often had DSM-IV ADHD symptoms. CONCLUSION: ADHD symptoms are as common among school children in Kinshasa as elsewhere. The socio-demographic factors described as risk factors for ADHD in high-income countries were not identified in this study. PMID- 16245987 TI - Gender differences among oral health care workers in caring for HIV/AIDS patients in Osun State, Nigeria. AB - The study investigated the relationship between gender and knowledge, attitude and practice of infection control among oral health care workers in the management of patients with HIV/AIDS in Osun State of Nigeria. It was a cross sectional survey using 85 oral Health care workers (OHCWs) enlisted in the public dental health clinics. A self-administered questionnaire was designed and used for data collection. A total of 85 questionnaires were distributed. The response rate was 93%; 42 (53%) were males and 37 (47%) females. The majority of the respondents were in the 25-40 year old age group and the mean age was 37.3 years. This study found significant differences in gender and ability to identify HIV/AIDS oral manifestations (p<0.001) and recognition of HIV/AIDS risk factors (p<0.001). There was statistically significant gender difference and infection control practices (p=0.02) among the OHCWs. Males were more compliant to the universal cross-infection control principle than the female respondents. A significant association (p< 0.001) was found between OHCW gender and their attitude to the management of HIV/AIDS patients with males showing a better attitude towards the care of HIV/AIDS patients. This study shows that there are significant gender difference in attitudes, behaviour and practices of OHCW with males faring better than the females. National AIDS Control Programme, Health Control bodies, Health educators and other organizations should make efforts to improve the attitude and practice of oral health care workers regarding the management of patients with HIV/AIDS. PMID- 16245988 TI - Gender power imbalance on women's capacity to negotiate self-protection against HIV/AIDS in Botswana and South Africa. AB - BACKGROUND: Gender power imbalance, which translates into a power imbalance in sexual interactions, is increasingly being recognized as a factor in fueling the spread of HIV/AIDS by increasing the number of unsafe sexual encounters. OBJECTIVES: To examine the influence of gender power imbalance and other factors on women's capacity to negotiate self protection against HIV infection; as well as men's response to the suggested condom use. METHODS: Drawing on data gathered from 2658 women aged 18-49 years in a cross-sectional survey in Kwa Zulu Natal Province of South Africa and Botswana, the study used descriptive statistics and logistic regression to reveal a number of gender related factors that significantly affect women's ability to protect themselves against HIV infection. RESULTS: Gender power imbalance significantly affects women's ability to suggest condom use to their partners. The study showed that it is women with partners 10 or more years older than them, abused women, and those economically dependent on their partners who are less likely to suggest condom use to their partners. Gender power imbalance also influences men's inclination towards refusing to use the suggested condom. The study showed that men are more likely to refuse to use the condom when the age difference between them and their female partners is wide, if they are in a married relationship, and where there is no communication about HIV/AIDS between them and their partners. What is more disturbing is the finding that it is men with multiple partners who are significantly more likely to refuse to use the condom. CONCLUSION: Across all levels of society, there is a need to see a social paradigm shift that transforms relationships between women and men, from the one of inequality and dominance as is the case in patriarchal societies, to equality, respect and consideration for one another. PMID- 16245990 TI - Incidence of HIV infection at the time of incident reporting, in victims of sexual assault, between 2000 and 2004, in Transkei, Eastern Cape, South Africa. AB - BACKGROUND: HIV/AIDS epidemic and sexual assault have emerged as the most serious public health problems in South Africa. The country has about 5-million HIV infected individuals. About a million women are raped every year. OBJECTIVES: To study the incidence of HIV infection in victims of sexual assaults. DESIGN: This is a retrospective descriptive study. There were 831 victims of sexual assault, who attended the Sinawe Centre during the 5-year (2000-2004) study period. Sinawe Centre is the only unit in this area which deals with cases of sexual assaults. Data were analyzed using PRISM R computer program. METHODS: This is a retrospective descriptive study. There were 831 victims of sexual assault, who attended the Sinawe Centre during the 5-year (2000-2004) study period. Sinawe Centre is the only unit in this area which deals with cases of sexual assaults. Data were analyzed using PRISM R computer program. RESULTS: Eight hundred and thirty one cases of sexual assaults (rape) were reported in last 5-years (2000 2004). Increase in sexual assaults by three-times, and one-and-half-fold increase in HIV seropositivity has recorded over the period of 5 years. In 2000, 76 (9.7%) sexual assault cases were registered, and in 2004, 237 (28.5%) cases. Of them, 443 (53.3%) were children under 15-years of age. A total of 114 (13.7%) tested positive for HIV, and 23 (2.7%) were children less than 15 years. In 2000, only 8 (10.5%) tested positive for HIV, and this had climbed to 39 (16.5%) in 2004. There is significant difference in HIV seropositivity among children compared to adults, p<0.05, and Paired t=3.45 with 3df. CONCLUSION: There is an increasing incidence of HIV infection among victims of sexual assaults in this study. PMID- 16245991 TI - Health system constraints to optimal coverage of the prevention of mother-to child HIV transmission programme in South Africa: lessons from the implementation of the national pilot programme. AB - BACKGROUND: It is three years since the government of South Africa began implementing a PMTCT programme. Over this period of time attempts have been made to scale up this programme across all provinces under routine health service conditions. OBJECTIVES: To report on the uptake and performance of South Africa's national pilot programme for preventing mother to child HIV transmission (PMTCT) and to identify health system constraints to optimal coverage. METHODS: Routine programme data were collected from antenatal records and delivery registers at the pilot sites and interviews were conducted with health workers on site and with provincial programme managers. RESULTS: Routine PMTCT programme data were collected from all 18 pilot sites for the period January to December 2002. During this period, of 84406 women attending the sites for first antenatal visits, 47267 (56%) agreed to an HIV test. 14340 (30%) of the women tested were HIV positive and of these 7853 (55%) were dispensed nevirapine. 7950 (99%) of infants born to women identified as being HIV positive received nevirapine syrup. 58% (4196/7237) of HIV positive women expressed an intention to exclusively formula feed, and 42% (3041/7237) intended to exclusively breastfeed. 1907 infants were due for 12 month HIV testing between January and December 2002, of these 949 (50%) infants were tested. CONCLUSION: Programme effectiveness was limited by the low rate of HIV test acceptance, poor delivery of nevirapine to mothers and inability to track mother-infant pairs postnatally for 12-month HIV testing of infants. Infant feeding intentions of mothers suggest inadequate counselling and possible negative effects of the provision of free formula milk. The poor performance of the main components of this programme will seriously reduce its operational effectiveness. There is a need for greater integration of VCT within antenatal care, a review of the current policy of providing free formula milk and an alternative model for mother-infant follow up. PMID- 16245989 TI - Socio-behaviour challenges to phase III HIV vaccine trials in Sub-Saharan Africa. AB - BACKGROUND: A number of countries in sub-Saharan Africa are preparing for HIV vaccine efficacy trials. Social and behavioural factors related to HIV transmission require examination in each setting where these trials are considered. As part of this, several countries have also recently begun preparatory research investigating relevant social and behavioural issues. There is a need for a review of the literature to help focus such research efforts in Sub-Saharan Africa. OBJECTIVES: To examine key social and behavioural issues that may impact on the conduct of HIV vaccine efficacy trials in sub-Saharan Africa. DESIGN: Literature review METHODS: Major databases (PubMed, PsychInfo, EBSCOhost, and AIDSline) were searched for literature that discussed social and behavioural issues related to HIV vaccine trials. Three areas are highlighted as being particularly significant for HIV vaccine research: (1) willingness to participate in future HIV vaccine efficacy trials, (2) retention of participants in studies, and (3) sexual risk reporting during trials. For each of these topics, major findings from both developed and developing countries are described and avenues for further research are discussed. RESULTS: There are few data from Sub-Saharan Africa regarding willingness to participate in HIV vaccine trials. Data on participant retention rates varies widely, and maintaining large cohorts of individuals within Phase III trials presents an important challenge. In addition, the possible impact of trial participation on sexual disinhibition, and response bias on sexual risk-reporting remain as issues for HIV vaccine trials in African contexts. CONCLUSION: Social and behavioural research forms an important part of preparations for HIV vaccine efficacy trials, and there is a clear need for more research of this type in Sub-Saharan Africa. Innovative approaches are required to address issues such as willingness to participate in vaccine research, participant retention during efficacy trials, and the accurate reporting by participants of sexual risk behaviours. PMID- 16245992 TI - Heart disease among children with HIV/AIDS attending the paediatric infectious disease clinic at Mulago Hospital. AB - BACKGROUND: There are very few published studies of heart disease in HIV infected children living in sub-Saharan Africa, a region with more than 50% of the world's population of HIV infected patients. OBJECTIVES: To determine the prevalence, and describe the type and clinical presentation of heart disease among children with HIV attending an ambulatory clinic. METHODOLOGY: Two hundred and thirty (230) HIV infected children attending the Paediatric Infectious Disease Clinic at Mulago hospital were recruited by simple random sampling in a cross-sectional study. The children were evaluated clinically, and investigated by electrocardiography and echocardiography. RESULTS: Thirty-two children (13.9%) had asymptomatic HIV disease, 156 (67.8%) had AIDS related complex while 42 (18.3%) had AIDS. Heart abnormalities were detected in 51% of the children (40.0% by echocardiography alone and 26.5% by electrocardiography alone). Heart abnormalities were most prevalent in children with AIDS (76.2%) and least prevalent in children with asymptomatic HIV disease (25.0%). The abnormalities included; Sinus tachycardia (21%), left ventricular systolic dysfunction (17%), right ventricular dilatation (14%), congenital heart disease (4.8%), dilated cardiomyopathy (3.0%), pericarditis (2.2%) and cor pulmonale (1.3%). Children with left ventricular systolic dysfunction significantly had easy fatigability, dyspnoea on exertion and tachypnoea. Other heart abnormalities presented with non-specific clinical features. CONCLUSION: Heart abnormalities were common especially in children with symptomatic HIV disease and included sinus tachycardia, left ventricular systolic dysfunction and right ventricular dilatation. The detected heart abnormalities, except left ventricular systolic dysfunction, had non-specific clinical features. PMID- 16245993 TI - Prevalence of oral diseases/conditions in Uganda. AB - OBJECTIVES: The aim was to report the prevalence of oral diseases/conditions among a Ugandan population. METHODS: Subjects aged 12 (n=696) and 35-44 years (n=396) were chosen from randomly selected urban and peri-urban areas of Arua, Mbale, Kampala and Mbarara districts. They were clinically examined by 4 trained and calibrated dentists for oral diseases/conditions using criteria described by World Health Organisation. RESULTS: Dental caries (DMFT >or=1) was recorded in 40% and 62.5% of the children and adults, respectively. The overall mean DMFT score was 0.9 for children and 3.4 for adults. Caries was significantly more severe in females as compared to males in children (p<0.05), whereas in adults, there was no significant gender difference. Kampala had a significantly higher mean DMFT score compared to other districts in all age groups (p>0.05). Culculus deposits were generally, more prevalent in adults as compare to children except in Mbarara district. Gum bleeding was also significantly more prevalent among children as compared to adults (p<0.05). Significantly higher prevalence of gum bleeding in both children and adults was recorded in Arua district as compared to other areas (p<0.05). Each of the age groups had a prevalence of malocclusion of 61%. However, the severity of malocclusion varied between age groups and districts. The prevalence of dental fluorosis was 3% and 4% for children and adults, respectively. All subjects in Arua district were fluorosis-free. Tetracycline enamel staining was less than 1% in both age groups. Enamel attrition was more prevalent in adults as compared to children: 19% versus 1%. CONCLUSION: The prevalence of oral diseases/conditions was generally low among the study population. Caries experience was significantly higher in the Kampala (urban) district as compared to rural districts in all age groups; the D - component being the major contributor. PMID- 16245994 TI - Helicobacter pylori and histopathological changes of gastric mucosa in Uganda population with varying prevalence of stomach cancer. AB - OBJECTIVES: Study the pathological changes in gastric mucosa of Nyarwanda, Nkole (both with high prevalence of stomach cancer) and Ganda (with low prevalence of this cancer) ethnic groups in the presence of Helicobacter pylori (H. pylori) infection. RESEARCH QUESTION: Do pathological changes accompanying H. pylori infection explain the varying prevalence of stomach cancer in these populations? DESIGN: Retrospective cross sectional study. SUBJECTS: A total of 114 patients of the above ethnic groups with upper gastrointestinal symptoms who underwent endoscopic biopsy examination between January 1996 and June 2002 formed the basis of this study. RESULTS: The severity of gastritis correlated with the presence of H. Pylori in Ganda and Nyarwanda but not in Nkole. Intestinal metaplasia (IM) was observed in Nyarwanda and Nkole and in some of these cases there was H. pylori. Gastric atrophy (GA) was also commonly observed in Nkole and Nyarwanda and H. pylori was detected more in the severe form of GA. Lymphoid follicle formation was not associated with H. pylori infection in all study groups. CONCLUSION: The major histological features relating stomach cancer to H. pylori in this study were presence of the infection in IM and GA that was observed mainly in Nyarwanda and Nkole. The lack of association between presence of lymphoid follicle and H. pylori infection probably explains the rarity of MALT lymphoma in Africa as these tumours are said to arise from H. pylori associated lymphoid follicles. PMID- 16245995 TI - Relationship between malaria infection intensity and rainfall pattern in Entebbe peninsula, Uganda. AB - BACKGROUND: This study was to assess malaria prevalence in relation to rainfall pattern in different localities of Entebbe Municipality, Uganda. A cross sectional study was conducted amongst the local community residing in the area from January 2003 to January 2004 to investigate the incidence and intensity of malaria infection. METHODS: Thick and thin blood smears were made from each patient with fever (body temperature >or= 37.5 degrees C). The slides were examined microscopically for malaria parasites. A total of 616 residents aged 2 to 50 years were registered in the study by name, age, sex and residential location. Spearman correlation coefficient (r) was used to evaluate relationship between parasite density, age and body temperature on one hand and rainfall plus parasite density on the other hand. RESULTS: A direct relationship was observed between malaria transmission and monthly rainfall in Entebbe Municipality. About 69.8% of the patients had fever. Parasite density fluctuated according to monthly rainfall pattern. Two peaks of high parasite density was observed, each peak coinciding with a peak rainfall pattern of the bimodal annual rain seasons. There was a negative but significant correlation (r = -0.09271; p < 0.0214) between parasite density and age, suggesting that mature individuals clear parasites more effectively than children. Furthermore, we observed a bimodal peak of mean parasite density in children and adults; peak in children is higher than of adults, each peak coinciding with rainfall pattern. There is also a significant positive correlation between parasite density and body temperature (r = 0.1927; p = 0.0001). However, there is no significant variation in mean parasite densities in the different locations of Entebbe Municipality. CONCLUSION: Our study confirms rainfall pattern and age influence parasite density and are important determinants of malaria infection and transmission in Entebbe Municipality. Increased parasite density in children is a useful indicator for monitoring intensity of infection. This information is valuable in policy formulation for control of malaria during periods of intense transmission. PMID- 16245996 TI - Crossing the digital divide: the contribution of information technology to the professional performance of malaria researchers in Africa. AB - BACKGROUND: The US National Library of Medicine supports the Multilateral Initiative on Malaria (MIM) through the design, implementation, and operation of the Multilateral Initiative on Malaria Communications Network (MIMCom.) MIMCom makes possible enhanced access to the Internet and to medical literature. OBJECTIVES: The main objectives of the present study were to examine the use of MIMCom supported information technology (IT) by scientists, students, and administrative personnel to facilitate communication, retrieve information, obtain documents, write proposals, and prepare papers for publication; and to determine the contribution of this intervention to their professional performance. METHODS: The authors analyzed the contribution of enhanced Internet connectivity and access to electronic information resources to the performance of malaria research staff and their institutes through a cross-sectional questionnaire survey of 181 respondents at 14 health research centers in Africa. Separate reviews of bandwidth usage, requests for document delivery, and publications in peer reviewed journals support the data of the survey. RESULTS: The MIMCom network makes a positive contribution to the performance of malaria researchers and support staff at the sites reviewed by improving e-mail exchange, access to published literature, and research proposal development and submission. Implications of these findings are discussed. CONCLUSION: By providing full access to the Internet and the resources of the WorldWide Web, MIMCom has been shown to be invaluable to malaria researchers and their institutes in Africa. This access has increased visibility of scientists in their respective institutions and provided opportunities for stronger engagement with the international scientific community. PMID- 16245997 TI - Natural history and epidemiology of post transplantation diabetes mellitus. AB - OBJECTIVE: High patient mortality continues to be the major threat to the success of solid organ transplantation despite improvements in the control of immunologic phenomena post-transplantation with improvements in grafts survival. Post transplantation diabetes mellitus (PTD) occurring commonly in patients receiving immunosuppressives has been identified as a major culprit. AIM: This paper reviews the current concepts of PTD. METHODS: A Medline literature search limited to research /review articles in English language, spanning 1984 January - 2004 June on post transplantation diabetes mellitus or hyperglycemia was done. RESULTS: Post-transplantation diabetes mellitus is a heterogeneous condition of abnormal glucose tolerance with a variable onset, duration and severity. It is defined as a fasting plasma glucose = 7.0 mmol / L (126 mg / dL) and / or a 2 hour plasma glucose during an OGTT = 11.1 mmol / L (200 mg / dL), or a casual plasma glucose = 11.1 mmol / L (200 mg / dL), on 3 or more occasions over a period of at least 3 months. The risk factors predisposing to PTD are age, recipient's ethnicity, immunosuppressive regimen and family history of diabetes. The condition not only increases the risk of graft-related complications such as graft rejection, graft loss and infection, but also is a major determinant of the increased cardiovascular morbidity and mortality. CONCLUSION: The most important risk factor predisposing to the development of PTD is the immunosuppressive drugs. The selection of immunosuppressive regimen should take into account individuals diabetes risk profile and the relative diabetogenicity and risk for diabetes of each immunosuppressant, balancing minimal risk of diabetes with effective immunosuppression. PMID- 16245998 TI - Extremity haemangiopericytoma, a case report from Nigeria. AB - Haemangiopericytoma is an uncommon soft tissue sarcoma of vascular origin. It occurs more frequently in the extremities than elsewhere in the body although it can arise in any organ. Wide surgical excision is the mainstay of treatment. However, adjuvant radiotherapy and chemotherapy are desirable because the malignant nature of this tumour is frequently unpredictable. Adjuvant therapy is recommended for metastases, recurrence and incomplete resection. Long term follow up is essential in all cases as recurrence can occur several years after treatment. Where little or no experience with managing this tumor exists, it is important to be aware of its clinical behaviour and the treatment options, hence this case reports. PMID- 16245999 TI - Ruptured false iliac artery aneurysm--a case report from Nigeria. AB - A 23 year old male Nigerian presented to the Nephrology unit of the Olabisi Onabanjo University Teaching Hospital, Sagamu, Nigeria with features of chronic renal failure secondary to chronic glomerulonephritis. He was referred to the surgical department of the hospital on account of severe torrential uncontrollable bleeding from an infected cannulation site. He was maintained on regular hemodialysis but developed femoral triangle sepsis at the cannulation site and aneurysm of the external iliac artery. The aneurysm later ruptured with severe jet exsanguinations bleeding. Exploration revealed a-4 cm rent in the external iliac artery that was sutured with 6/0 prolene, interrupted, using clamp and suture technique. Perfusion was satisfactory in the immediate post-operative period. The patient was transfused with 11 pints of blood within 12 hours and had an uneventful post-operative period. PMID- 16246000 TI - Subtotal amelia in a child with autosomal recessive hypohidrotic ectodermal dysplasia. AB - We report an inbred Tunisian family, in which the proband manifested signs of hypohidrotic ectodermal dysplasia, subtotal amelia, scoliosis and left renal agenesis. Two other family members had the full clinical criteria of hypohidrotic ectodermal dysplasia, characterized by deficient sweat glands, hypodontia, hypoplasia of the mucous glands, and fine hair. Nine family subjects had variable clinical expression of the disorder. PMID- 16246001 TI - Paradoxical helminthiasis and giardiasis in Cape Town, South Africa: epidemiology and control. AB - BACKGROUND: South Africa has endorsed a World Health Assembly (WHA) resolution calling for control of soil-transmitted helminths (STHs). In Cape Town, services and housing that exist in old-established suburbs should minimise the prevalence of intestinal parasitic infections, even when residents are poor. Where families live in shacks in densely-populated areas without effective sanitation, more than 90% of children can be infected by STHs. The humoral immune response to worms theoretically favours infection by Mycobacterium tuberculosis and HIV. OBJECTIVES: Obtain estimates of gender-, age-, school-related and overall prevalence of helminthiasis and giardiasis in a low-income but well-serviced community. Assess possible sources of infection. Alert health services to the need for control measures and the threat from protozoal pathogens. Warn that the immune response to intestinal parasites may favour tuberculosis (TB) and HIV/AIDS. METHODS: A cross-sectional study of the prevalence of helminthiasis and giardiasis was carried out in a large, non-selective sample of children attending nine schools. Gender, school and age effects were related to non-medical preventive services, sewage disposal practices and possible sources of infection. RESULTS: The overall STH infestation rate was 55.8%. Prevalence was inluenced by school and age but not by gender. Eggs and cysts were seen at the following prevalence: Ascaris 24.8%; Trichuris 50.6%; Hymenolepis nana 2.2%; Enterobius 0.6%; Giardia 17.3%; hookworm 0.08%; and Trichostrongylus 0.1%. Approximately 60% of sewage sludge is used in a form that will contain viable eggs and cysts. CONCLUSION: Prevalence trends in this old community in Cape Town could indicate infection by swallowing eggs or cysts on food or in water, more than by exposure to polluted soil. Sewage sludge and effluent might be sources of infection. In adjacent, under serviced, newer communities, promiscuous defecation occurs. Probable vectors are discussed. The immune response to intestinal parasites might be a risk factor for HIV/AIDS and TB. PMID- 16246005 TI - Rho GTPases and the control of cell behaviour. AB - Rho, Rac and Cdc42, three members of the Rho family of small GTPases, each control a signal transduction pathway linking membrane receptors to the assembly and disassembly of the actin cytoskeleton and of associated integrin adhesion complexes. Rho regulates stress fibre and focal adhesion assembly, Rac regulates the formation of lamellipodia protrusions and membrane ruffles, and Cdc42 triggers filopodial extensions at the cell periphery. These observations have led to the suggestion that wherever filamentous actin is used to drive a cellular process, Rho GTPases are likely to play an important regulatory role. Rho GTPases have also been reported to control other cellular activities, such as the JNK (c Jun N-terminal kinase) and p38 MAPK (mitogen-activated protein kinase) cascades, an NADPH oxidase enzyme complex, the transcription factors NF-kappaB (nuclear factor kappaB) and SRF (serum-response factor), and progression through G1 of the cell cycle. Thus Rho, Rac and Cdc42 can regulate the actin cytoskeleton and gene transcription to promote co-ordinated changes in cell behaviour. We have been analysing the biochemical contributions of Rho GTPases in cell movement and have found that Rac controls cell protrusion, while Cdc42 controls cell polarity. PMID- 16246004 TI - Toxoplasma gondii acyl-lipid metabolism: de novo synthesis from apicoplast generated fatty acids versus scavenging of host cell precursors. AB - Toxoplasma gondii is an obligate intracellular parasite that contains a relic plastid, called the apicoplast, deriving from a secondary endosymbiosis with an ancestral alga. Metabolic labelling experiments using [14C]acetate led to a substantial production of numerous glycero- and sphingo-lipid classes in extracellular tachyzoites. Syntheses of all these lipids were affected by the herbicide haloxyfop, demonstrating that their de novo syntheses necessarily required a functional apicoplast fatty acid synthase II. The complex metabolic profiles obtained and a census of glycerolipid metabolism gene candidates indicate that synthesis is probably scattered in the apicoplast membranes [possibly for PA (phosphatidic acid), DGDG (digalactosyldiacylglycerol) and PG (phosphatidylglycerol)], the endoplasmic reticulum (for major phospholipid classes and ceramides) and mitochondria (for PA, PG and cardiolipid). Based on a bioinformatic analysis, it is proposed that apicoplast produced acyl-ACP (where ACP is acyl-carrier protein) is transferred to glycerol-3-phosphate for apicoplast glycerolipid synthesis. Acyl-ACP is also probably transported outside the apicoplast stroma and irreversibly converted into acyl-CoA. In the endoplasmic reticulum, acyl-CoA may not be transferred to a three-carbon backbone by an enzyme similar to the cytosolic plant glycerol-3-phosphate acyltransferase, but rather by a dual glycerol-3-phosphate/dihydroxyacetone-3-phosphate acyltransferase like in animal and yeast cells. We further showed that intracellular parasites could also synthesize most of their lipids from scavenged host cell precursors. The observed appearance of glycerolipids specific to either the de novo pathway in extracellular parasites (unknown glycerolipid 1 and the plant like DGDG), or the intracellular stages (unknown glycerolipid 8), may explain the necessary coexistence of both de novo parasitic acyl-lipid synthesis and recycling of host cell compounds. PMID- 16246006 TI - The efficiency and plasticity of mitochondrial energy transduction. AB - Since it was first realized that biological energy transduction involves oxygen and ATP, opinions about the amount of ATP made per oxygen consumed have continually evolved. The coupling efficiency is crucial because it constrains mechanistic models of the electron-transport chain and ATP synthase, and underpins the physiology and ecology of how organisms prosper in a thermodynamically hostile environment. Mechanistically, we have a good model of proton pumping by complex III of the electron-transport chain and a reasonable understanding of complex IV and the ATP synthase, but remain ignorant about complex I. Energy transduction is plastic: coupling efficiency can vary. Whether this occurs physiologically by molecular slipping in the proton pumps remains controversial. However, the membrane clearly leaks protons, decreasing the energy funnelled into ATP synthesis. Up to 20% of the basal metabolic rate may be used to drive this basal leak. In addition, UCP1 (uncoupling protein 1) is used in specialized tissues to uncouple oxidative phosphorylation, causing adaptive thermogenesis. Other UCPs can also uncouple, but are tightly regulated; they may function to decrease coupling efficiency and so attenuate mitochondrial radical production. UCPs may also integrate inputs from different fuels in pancreatic beta-cells and modulate insulin secretion. They are exciting potential targets for treatment of obesity, cachexia, aging and diabetes. PMID- 16246007 TI - Lipid interactions with bacterial channels: fluorescence studies. AB - Interactions between a membrane protein and the lipid molecules that surround it in the membrane are important in determining the structure and function of the protein. These interactions can be pictured at the molecular level using fluorescence spectroscopy, making use of the ability to introduce tryptophan residues into regions of interest in bacterial membrane proteins. Fluorescence quenching methods have been developed to study lipid binding separately on the two sides of the membrane. Lipid binding to the surface of the mechanosensitive channel MscL is heterogeneous, with a hot-spot for binding anionic lipid on the cytoplasmic side, associated with a cluster of three positively charged residues. The environmental sensitivity of tryptophan fluorescence emission has been used to identify the residues at the ends of the hydrophobic core of the second transmembrane alpha-helix in MscL. The efficiency of hydrophobic matching between MscL and the surrounding lipid bilayer is high. Fluorescence quenching methods can also be used to study binding of lipids to non-annular sites such as those between monomers in the homotetrameric potassium channel KcsA. PMID- 16246008 TI - Simulation studies of the interactions between membrane proteins and detergents. AB - Interactions between membrane proteins and detergents are important in biophysical and structural studies and are also biologically relevant in the context of folding and transport. Despite a paucity of high-resolution data on protein-detergent interactions, novel methods and increased computational power enable simulations to provide a means of understanding such interactions in detail. Simulations have been used to compare the effect of lipid or detergent on the structure and dynamics of membrane proteins. Moreover, some of the longest and most complex simulations to date have been used to observe the spontaneous formation of membrane protein-detergent micelles. Common mechanistic steps in the micelle self-assembly process were identified for both alpha-helical and beta barrel membrane proteins, and a simple kinetic mechanism was proposed. Recently, simplified (i.e. coarse-grained) models have been utilized to follow long timescale transitions in membrane protein-detergent assemblies. PMID- 16246009 TI - Spectroscopic studies of phospholamban variants in phospholipid bilayers. AB - Phospholamban (PLB) is a 52 amino acid transmembrane protein found in the sarcoplasmic reticulum of cardiac myocytes, where it regulates the transport of calcium ions by SERCA (sarcoplasmic/endoplasmic reticulum Ca2+-ATPase). This work has shown that the cytoplasmic domain of PLB associates with phospholipid vesicles, possibly with the lipid polar head groups, and, in doing so, undergoes a transition from a random coil to a more helical conformation. These findings support a recent hypothesis that the cytoplasmic domain of PLB acts as a conformational switch, alternating between an orientation that lies across the membrane surface and an upright orientation that associates with the regulatory site of SERCA. PMID- 16246010 TI - Molecular simulations and lipid-protein interactions: potassium channels and other membrane proteins. AB - Molecular dynamics simulations may be used to probe the interactions of membrane proteins with lipids and with detergents at atomic resolution. Examples of such simulations for ion channels and for bacterial outer membrane proteins are described. Comparison of simulations of KcsA (an alpha-helical bundle) and OmpA (a beta-barrel) reveals the importance of two classes of side chains in stabilizing interactions with the head groups of lipid molecules: (i) tryptophan and tyrosine; and (ii) arginine and lysine. Arginine residues interacting with lipid phosphate groups play an important role in stabilizing the voltage-sensor domain of the KvAP channel within a bilayer. Simulations of the bacterial potassium channel KcsA reveal specific interactions of phosphatidylglycerol with an acidic lipid-binding site at the interface between adjacent protein monomers. A combination of molecular modelling and simulation reveals a potential phosphatidylinositol 4,5-bisphosphate-binding site on the surface of Kir6.2. PMID- 16246011 TI - Binding sites of lipophilic quinone and quinone analogue inhibitors in the cytochrome b6f complex of oxygenic photosynthesis. AB - The main structural features of the cytochrome b6f complex, solved to 3.0-3.1 A (1 A = 10(-10) m) in the cyanobacterium Mastigocladus laminosus and the green alga Chlamydomonas reinhardtii are discussed. The discussion is focused on the binding sites of plastoquinone and quinone analogue inhibitors discerned in the structure. These sites mark the pathway(s) of quinone translocation across the complex. PMID- 16246012 TI - Lipids in and around photosynthetic reaction centres. AB - Reaction centres are membrane-embedded pigment-protein complexes that transduce the energy of sunlight into a biologically useful form. The most heavily studied reaction centres are the PS-I (Photosystem I) and PS-II complexes from oxygenic phototrophs, and the reaction centre from purple photosynthetic bacteria. A great deal is known about the compositions and structures of these reaction centres, and the mechanism of light-activated transmembrane electron transfer, but less is known about how they interact with other components of the photosynthetic membrane, including the membrane lipids. X-ray crystallography has provided high resolution structures for PS-I and the purple bacterial reaction centre, and revealed binding sites for a number of lipids, either embedded in the protein interior or attached to the protein surface. These lipids play a variety of roles, including the binding of cofactors and the provision of structural support. The challenges of modelling surface-associated electron density features such as lipids, detergents, small amphiphiles and ions are discussed. PMID- 16246013 TI - Quantification of the effects of melittin on liposome structure. AB - An optical technique, dual-polarization interferometry, has been used to examine lipid structures at the solid/liquid interface. Changes in the lipid structures, in real time, were examined as a consequence of challenging them with a peptide (melittin) that is known to induce liposome rupture. This work suggests that it should be possible to obtain a better understanding of the detail of the melittin rupture process. PMID- 16246014 TI - Reaction mechanism and phospholipid structures of bovine heart cytochrome c oxidase. AB - Bovine heart cytochrome c oxidase is a large multi-component membrane protein containing several phospholipids. X-ray structures of this enzyme at high resolution, determined recently, show a trigonal planar structure of CuB site in the O2 reduction site, which could contribute critically to the four-electron reduction of O2 bound at haem a3, and a hydrogen bond network, through which the proton pump is driven by haem a. The possible roles of phospholipids in the enzyme functions are discussed. PMID- 16246015 TI - Specific protein-lipid interactions in membrane proteins. AB - Many membrane proteins selectively bind defined lipid species. This specificity has an impact on correct insertion, folding, structural integrity and full functionality of the protein. How are these different tasks achieved? Recent advances in structural research of membrane proteins provide new information about specific protein-lipid interactions. Tightly bound lipids in membrane protein structures are described and general principles of the binding interactions are deduced. Lipid binding is stabilized by multiple non-covalent interactions from protein residues to lipid head groups and hydrophobic tails. Distinct lipid-binding motifs have been identified for lipids with defined head groups in membrane protein structures. The stabilizing interactions differ between the electropositive and electronegative membrane sides. The importance of lipid binding for vertical positioning and tight integration of proteins in the membrane, for assembly and stabilization of oligomeric and multisubunit complexes, for supercomplexes, as well as for functional roles are pointed out. PMID- 16246016 TI - Organ specificity in the circadian control of plant gene expression. AB - Of the many plant genes whose expressions are controlled by the circadian clock, one of the phosphoenolpyruvate carboxylase kinase genes in soya bean (Glycine max) exhibits the unusual property that its control is organ-specific--it is under circadian control in leaves but not in roots. Preliminary experiments suggest that the same is true for at least one gene in Arabidopsis thaliana. It will be important to define the extent and function of this phenomenon and the underlying mechanism. PMID- 16246017 TI - The co-ordination of central plant metabolism by the circadian clock. AB - A circadian clock optimizes many aspects of plant biology relative to the light/dark cycle. One example is the circadian control of primary metabolism and CO2 fixation in plants that carry out a metabolic adaptation of photosynthesis called CAM (crassulacean acid metabolism). These plants perform primary CO2 fixation at night using the enzyme phosphoenolpyruvate carboxylase and exhibit a robust rhythm of CO2 fixation under constant conditions. Transcriptomic analysis has revealed that many genes encoding enzymes in primary metabolic pathways such as glycolysis and starch metabolism are under the control of the circadian clock in CAM plants. These transcript changes are accompanied by changes in metabolite levels associated with flux through these pathways. The molecular basis for the circadian control of CAM remains to be elucidated. Current research is focusing on the identity of the CAM central oscillator and the output pathway that links the central oscillator to the control of plant metabolism. PMID- 16246018 TI - Synchronizing the Neurospora crassa circadian clock with the rhythmic environment. AB - The metronomic predictability of the environment has elicited strong selection pressures for the evolution of endogenous circadian clocks. Circadian clocks drive molecular and behavioural rhythms that approximate the 24 h periodicity of our environment. Found almost ubiquitously among phyla, circadian clocks allow preadaptation to rhythms concomitant with the natural cycles of the Earth. Cycles in light intensity and temperature for example act as important cues that couple circadian clocks to the environment via a process called entrainment. This review summarizes our current understanding of the general and molecular principles of entrainment in the model organism Neurospora crassa, a simple eukaryote that has one of the best-studied circadian systems and light-signalling pathways. PMID- 16246019 TI - Degradation of the Neurospora circadian clock protein FREQUENCY through the ubiquitin-proteasome pathway. AB - Phosphorylation of the Neurospora circadian clock protein FREQUENCY (FRQ) promotes its degradation through the ubiquitin-proteasome pathway. Ubiquitination of FRQ requires FWD-1 (F-box/WD-40 repeat-containing protein-1), which is the substrate-recruiting subunit of an SCF (SKP/Cullin/F-box)-type ubiquitin ligase. In the fwd-1 mutant strains, FRQ degradation is defective, resulting in the accumulation of hyperphosphorylated FRQ and the loss of the circadian rhythmicities. The CSN (COP9 signalosome) promotes the function of SCF complexes in vivo. But in vitro, deneddylation of cullins by CSN inhibits SCF activity. In Neurospora, the disruption of the csn-2 subunit impairs FRQ degradation and compromises the normal circadian functions. These defects are due to the dramatically reduced levels of FWD-1 in the csn-2 mutant, a result of its rapid degradation. Other components of the SCF(FWD-1) complex, SKP-1 and CUL-1 are also unstable in the mutant. These results establish important roles for SCF(FWD-1) and CSN in the circadian clock of Neurospora and suggest that they are conserved components of the eukaryotic circadian clocks. In addition, these findings resolve the CSN paradox and suggest that the major function of CSN is to maintain the stability of SCF ubiquitin ligases in vivo. PMID- 16246020 TI - Organization of endogenous clocks in insects. AB - Insect and mammalian circadian clocks show striking similarities. They utilize homologous clock genes, generating self-sustained circadian oscillations in distinct master clocks of the brain, which then control rhythmic behaviour. The molecular mechanisms of rhythm generation were first uncovered in the fruit fly Drosophila melanogaster, whereas cockroaches were among the first animals where the brain master clock was localized. Despite many similarities, there exist obvious differences in the organization and functioning of insect master clocks. These similarities and differences are reviewed on a molecular and anatomical level. PMID- 16246021 TI - Zebrafish circadian clocks: cells that see light. AB - In the classical view of circadian clock organization, the daily rhythms of most organisms were thought to be regulated by a central, 'master' pacemaker, usually located within neural structures of the animal. However, with the results of experiments performed in zebrafish, mammalian cell lines and, more recently, mammalian tissues, this view has changed to one where clock organization is now seen as being highly decentralized. It is clear that clocks exist in the peripheral tissues of animals as diverse as Drosophila, zebrafish and mammals. In the case of Drosophila and zebrafish, these tissues are also directly light responsive. This light sensitivity and direct clock entrainability is also true for zebrafish cell lines and early-stage embryos. Using luminescent reporter cell lines containing clock gene promoters driving the expression of luciferase and single-cell imaging techniques, we have been able to show how each cell responds rapidly to a single light pulse by being shifted to a common phase, equivalent to the early day. This direct light sensitivity might be related to the requirement for light in these cells to activate the transcription of genes involved in DNA repair. It is also clear that the circadian clock in zebrafish regulates the timing of the cell cycle, demonstrating the wide impact that this light sensitivity and daily rhythmicity has on the biology of zebrafish. PMID- 16246023 TI - Implications of a genomic search for autophagy-related genes in trypanosomatids. AB - Autophagy is the process by which cellular components are directed to and degraded in the vacuole or lysosome and has been studied largely in yeasts. We present here an in silico genomic analysis of trypanosomatid autophagy aimed at highlighting similarities and differences with autophagy in other organisms. Less than half of the yeast autophagy-related proteins examined have certain putative orthologues in trypanosomatids. A cytosol-to-vacuole transport system is clearly lacking in these organisms. Other absences are even more unexpected and have implications for our understanding of the molecular mechanisms of autophagy. The results are consistent with taxon-specific addition of components to a core autophagy machinery during evolution. PMID- 16246022 TI - The extraordinary mitochondrion and unusual citric acid cycle in Trypanosoma brucei. AB - African trypanosomes are parasitic protozoa that cause sleeping sickness and nagana. Trypanosomes are not only of scientific interest because of their clinical importance, but also because these protozoa contain several very unusual biological features, such as their specially adapted mitochondrion and the compartmentalization of glycolytic enzymes in glycosomes. The energy metabolism of Trypanosoma brucei differs significantly from that of their hosts and changes drastically during the life cycle. Despite the presence of all citric acid cycle enzymes in procyclic insect-stage T. brucei, citric acid cycle activity is not used for energy generation. Recent investigations on the influence of substrate availability on the type of energy metabolism showed that absence of glycolytic substrates did not induce a shift from a fermentative metabolism to complete oxidation of substrates. Apparently, insect-stage T. brucei use parts of the citric acid cycle for other purposes than for complete degradation of mitochondrial substrates. Parts of the cycle are suggested to be used for (i) transport of acetyl-CoA units from the mitochondrion to the cytosol for the biosynthesis of fatty acids, (ii) degradation of proline and glutamate to succinate, (iii) generation of malate, which can then be used for gluconeogenesis. Therefore the citric acid cycle in trypanosomes does not function as a cycle. PMID- 16246024 TI - Post-genomic views of a 'unique' metabolism in the eukaryotic flagellum. AB - This short review summarizes recent advances in our understanding of energy metabolism within the eukaryotic flagellum. Using the example of adenylate kinase, we discuss how a requirement to target metabolic enzymes into the flagellum results in the presence of genes encoding novel isoforms of ubiquitous enzymes within flagellate eukaryotes. PMID- 16246025 TI - Plasmodium falciparum possesses organelle-specific alpha-keto acid dehydrogenase complexes and lipoylation pathways. AB - The human malaria parasite Plasmodium falciparum possesses a single mitochondrion and a plastid-like organelle called the apicoplast. Both organelles contain members of the KADH (alpha-keto acid dehydrogenase) complexes--multienzyme complexes that are involved in intermediate metabolism. In the asexual blood stage forms of the parasites, the alpha-ketoglutarate dehydrogenase and branched chain KADH complexes are both located in the mitochondrion, whereas the pyruvate dehydrogenase is exclusively found in the apicoplast. In agreement with this distribution, Plasmodium parasites have two separate and organelle-specific pathways that guarantee lipoylation of the KADH complexes in both organelles. A biosynthetic pathway comprised of lipoic acid synthase and lipoyl (octanoyl) ACP:protein Nepsilon-lipoyltransferase B is present in the apicoplast, whereas the mitochondrion is supplied with exogenous lipoic acid, and ligation of the metabolite to the KADH complexes is accomplished by a lipoate protein ligase A similar to that of bacteria and plants. Both pathways are excellent potential targets for the design of new antimalarial drugs. PMID- 16246026 TI - Maintaining the protective variant surface glycoprotein coat of African trypanosomes. AB - The African trypanosome Trypanosoma brucei has a precarious existence as an extracellular parasite of the mammalian bloodstream, where it is faced with continuous immune attack. Key to survival is a dense VSG (variant surface glycoprotein) coat, which is repeatedly switched during the course of a chronic infection. New data demonstrate a link between VSG synthesis and cell cycle progression, indicating that VSG is monitored during the trypanosome cell cycle. PMID- 16246027 TI - The myo-inositol-1-phosphate synthase gene is essential in Trypanosoma brucei. AB - The de novo synthesis of myo-inositol occurs via a two-step process: first, glucose 6-phosphate is converted into inositol 1-phosphate by an INO1 (myo inositol-1-phosphate synthase; EC 5.5.1.4); then, it is dephosphorylated by an inositol monophosphatase. The myo-inositol can then be incorporated into PI (phosphatidylinositol), which is utilized in a variety of cellular functions, including the biosynthesis of GPI (glycosylphosphatidylinositol) anchors. A putative INO1 was identified in the Trypanosoma brucei genome database and, by recombinant expression in Escherichia coli, was shown to be a catalytically active INO1. To investigate the importance of INO1, we created a conditional knockout, which, under non-permissive conditions, showed that INO1 is an essential gene in bloodstream form T. brucei and that the de novo synthesized myo inositol is used for the formation of PI and GPI anchors. PMID- 16246028 TI - What the genome sequence is revealing about trypanosome antigenic variation. AB - African trypanosomes evade humoral immunity through antigenic variation, whereby they switch expression of the gene encoding their VSG (variant surface glycoprotein) coat. Switching proceeds by duplication of silent VSG genes into a transcriptionally active locus. The genome project has revealed that most of the silent archive consists of hundreds of subtelomeric VSG tandem arrays, and that most of these are not functional genes. Precedent suggests that they can contribute combinatorially to the formation of expressed, functional genes through segmental gene conversion. These findings from the genome project have major implications for evolution of the VSG archive and for transmission of the parasite in the field. PMID- 16246030 TI - Crystallographic and single-particle analyses of native- and nucleotide-bound forms of the cystic fibrosis transmembrane conductance regulator (CFTR) protein. AB - Cystic fibrosis, one of the major human inherited diseases, is caused by defects in the CFTR (cystic fibrosis transmembrane conductance regulator), a cell membrane protein. CFTR acts as a chloride channel which can be opened by ATP. Low resolution structural studies of purified recombinant human CFTR are described in the present paper. Localization of the C-terminal decahistidine tag in CFTR was achieved by Ni2+-nitriloacetate nanogold labelling, followed by electron microscopy and single-particle analysis. The presence of the gold label appears to improve the single-particle-alignment procedure. Projection structures of CFTR from two-dimensional crystals analysed by electron crystallography displayed two alternative conformational states in the presence of nucleotide and nanogold, but only one form of the protein was observed in the quiescent (nucleotide-free) state. PMID- 16246029 TI - A molecular understanding of the catalytic cycle of the nucleotide-binding domain of the ABC transporter HlyB. AB - The ABC transporter (ATP-binding-cassette transporter) HlyB (haemolysin B) is the central element of a type I secretion machinery, dedicated to the secretion of the toxin HlyA in Escherichia coli. In addition to the ABC transporter, two other indispensable elements are necessary for the secretion of the toxin across two membranes in a single step: the transenvelope protein HlyD and the outer membrane protein TolC. Despite the fact that the hydrolysis of ATP by HlyB fuels secretion of HlyA, the essential features of the underlying transport mechanism remain an enigma. Similar to all other ABC transporters, ranging from bacteria to man, HlyB is composed of two NBDs (nucleotide-binding domains) and two transmembrane domains. Here we summarize our detailed biochemical, biophysical and structural studies aimed at an understanding of the molecular principles of how ATP hydrolysis is coupled to energy transduction, including the conformational changes occurring during the catalytic cycle, leading to substrate transport. We have obtained individual crystal structures for each single ground state of the catalytic cycle. From these and other biochemical and mutational studies, we shall provide a detailed molecular picture of the steps governing intramolecular communication and the utilization of chemical energy, due to ATP hydrolysis, in relation to resulting structural changes within the NBD. These data will be summarized in a general model to explain how these molecular machines achieve translocation of molecules across biological membranes. PMID- 16246031 TI - ABC proteins and antibiotic drug resistance: is it all about transport? AB - The precise mechanism of antibiotic-resistance-conferring ABC (ATP-binding cassette) proteins (termed NBD2) remains open to debate. Currently, two hypotheses are recognized. In one, the NBD2 proteins are envisaged to act at the ribosome to impair antibiotic access to the target site on the 23 S rRNA. In the other, NBD2 proteins are believed to act as the components of ATP driven efflux pumps by associating with membrane spanning proteins capable of binding and transporting antibiotics. Pertinent data in support of these two hypotheses are discussed in this paper. PMID- 16246032 TI - Control of the CFTR channel's gates. AB - Unique among ABC (ATP-binding cassette) protein family members, CFTR (cystic fibrosis transmembrane conductance regulator), also termed ABCC7, encoded by the gene mutated in cystic fibrosis patients, functions as an ion channel. Opening and closing of its anion-selective pore are linked to ATP binding and hydrolysis at CFTR's two NBDs (nucleotide-binding domains), NBD1 and NBD2. Isolated NBDs of prokaryotic ABC proteins form homodimers upon binding ATP, but separate after hydrolysis of the ATP. By combining mutagenesis with single-channel recording and nucleotide photolabelling on intact CFTR molecules, we relate opening and closing of the channel gates to ATP-mediated events in the NBDs. In particular, we demonstrate that two CFTR residues, predicted to lie on opposite sides of its anticipated NBD1-NBD2 heterodimer interface, are energetically coupled when the channels open but are independent of each other in closed channels. This directly links ATP-driven tight dimerization of CFTR's cytoplasmic NBDs to opening of the ion channel in the transmembrane domains. Evolutionary conservation of the energetically coupled residues in a manner that preserves their ability to form a hydrogen bond argues that this molecular mechanism, involving dynamic restructuring of the NBD dimer interface, is shared by all members of the ABC protein superfamily. PMID- 16246033 TI - Similarities between ATP-dependent and ion-coupled multidrug transporters. AB - The movement of drugs across biological membranes is mediated by two major classes of membrane transporters. Primary-active, ABC (ATP-binding cassette) multidrug transporters are dependent on ATP-binding/hydrolysis, whereas secondary active multidrug transporters are coupled to the proton (or sodium)-motive force that exists across the plasma membrane. Recent work on LmrA, an ABC multidrug transporter in Lactococcus lactis, suggests that primary- and secondary-active multidrug transporters share functional and structural features. Some of these similarities and their implications for the mechanism of transport by ABC multidrug transporters will be discussed. PMID- 16246034 TI - Do protein-lipid interactions determine the recognition of transmembrane helices at the ER translocon? AB - Membrane-protein integration, folding and assembly processes in vivo depend on complex targeting, translocation, chaperoning, and sorting machineries that somehow read the 'molecular code' built into the nascent polypeptide, ultimately producing a properly folded protein integrated into the correct target membrane. Although the main molecular constituents and the basic mechanistic principles of many of these machines are known in outline, the codes remain poorly defined and there is little quantitative information on how protein sequence affects the final structure of membrane proteins. By carefully designing model protein constructs, we have derived the first true biological hydrophobicity scale and have been able to get a first impression of how the position of a given type of residue within a transmembrane segment affects its ability to promote membrane insertion. PMID- 16246035 TI - Pathways for protein transport to seed storage vacuoles. AB - Plant vacuoles have multiple functions: they can act both as digestive organelles and as receptacles for storage proteins. Different types of vacuoles can coexist in the same cell, which adds complexity to the process of targeting to these compartments. A fuller understanding of this process is of evident value when endeavouring to exploit the plant secretory pathway for heterologous protein production. Positive sorting signals are required in order to sort proteins to vacuoles, and these have been split into three groups: ctVSS [C-terminal VSS (vacuolar sorting signals)], ssVSS (sequence-specific VSS) and physical structure VSS. The current working model posits that soluble proteins are delivered from the Golgi apparatus to the lytic vacuoles in clathrin-coated vesicles by virtue of their ssVSS, or to the storage vacuole [PSV (protein-storage vacuole)] in dense vesicles in a manner dependent on ctVSS or physical structure VSS. Although targeting to LV appears to be receptor-mediated, no such receptor has been identified for the recruitment of proteins to the PSV. We have studied the vacuolar targeting of two castor bean (Ricinus communis L.) storage proteins, proricin and pro 2 S albumin, in their native endosperm and in the heterologous system of tobacco protoplasts. We have found that both these proteins contain bona fide ssVSS and bind to sorting receptors in vitro in a similarly sequence specific manner. The apparent similarities to lytic VSS and possible implications with respect to the working model for transport to storage vacuoles are discussed. PMID- 16246036 TI - Protein import into mitochondria. AB - Mitochondria comprise approx. 1000-3000 different proteins, almost all of which must be imported from the cytosol into the organelle. So far, six complex molecular machines, protein translocases, were identified that mediate this process. The TIM23 complex is a major translocase in the inner mitochondrial membrane. It uses two energy sources, namely membrane potential and ATP, to facilitate preprotein translocation across the inner membrane and insertion into the inner membrane. Recent research has led to the discovery of a number of new constituents of the TIM23 complex and to the unravelling of the mechanisms of preprotein translocation. PMID- 16246037 TI - The complexity of pathways for protein import into thylakoids: it's not easy being green. AB - Numerous proteins are transported into or across the chloroplast thylakoid membrane. To date, two major pathways have been identified for the transport of luminal proteins (the Sec- and Tat-dependent pathways) and it is now clear that these protein translocases use fundamentally different transport mechanisms. Integral membrane proteins are inserted by means of at least two further pathways. One involves the input of numerous targeting factors, including SRP (signal recognition particle), FtsY and Albino3. Surprisingly, the other pathway does not involve any of the known chloroplastic targeting factors, and insertion is energy-independent, raising the possibility of an unusual 'spontaneous' insertion mechanism. PMID- 16246038 TI - Leptin: a potential cognitive enhancer? AB - It is well documented that the hormone leptin signals information regarding the status of fat stores to hypothalamic nuclei, which in turn control feeding behaviour and body weight. However, leptin and its receptor are widely expressed in many extra-hypothalamic brain regions, including hippocampus, brain stem and cerebellum. Moreover, evidence is accumulating that leptin has other neuronal functions that are unrelated to its effects on energy homeostasis. Indeed a role for leptin in neuronal development has been suggested as leptin-deficient rodents display abnormal brain development and leptin actively participates in the development of the hypothalamus. In the hippocampus, leptin is a potential cognitive enhancer as genetically obese rodents with dysfunctional leptin receptors display impairments in hippocampal synaptic plasticity. Moreover, direct administration of leptin into the hippocampus can facilitate hippocampal LTP (long-term potentiation) in vivo and improve memory processing in mice. At the cellular level, we have also shown that leptin has the capacity to convert short-term potentiation into LTP. Here, we review the data that leptin influences hippocampal synaptic plasticity via enhancing NMDA (N-methyl-D-aspartate) receptor function. We also provide evidence that rapid trafficking of NMDA receptors to the plasma membrane may underlie the effects of leptin on excitatory synaptic strength. PMID- 16246039 TI - Insulin and cholesterol pathways in neuronal function, memory and neurodegeneration. AB - Insulin and cholesterol play important roles in basic metabolic processes in peripheral tissues. Both insulin and cholesterol can also act as signalling molecules in the central nervous system that participate in neuronal function, memory and neurodegenerative diseases. A high-cholesterol diet improves spatial memory in experimental animals. beta-Amyloid, the toxic peptide in neurons of AD (Alzheimer's disease) patients, binds cholesterol and catalyses its oxidation to 7beta-hydroxycholesterol, a highly toxic oxysterol that is a potent inhibitor of alpha-PKC (alpha-protein kinase C), an enzyme critical in memory consolidation and synaptic plasticity and implicated in AD. Oxidized cholesterol also can act as a second messenger for insulin. Oxidized low-density lipoprotein inhibits insulin-dependent phosphorylation of the signalling kinases ERK (extracellular signal-regulated kinase) and PKB/Akt. In sporadic AD patients, insulin levels are decreased, suggesting links between AD and diabetes. Insulin signalling is also important in synaptic plasticity. Insulin receptors are up-regulated and undergo translocation after spatial learning. Insulin modulates the activity of excitatory and inhibitory receptors including the glutamate and gamma aminobutyric acid receptors and activates two biochemical pathways: the shc-ras mitogen-activated protein kinase pathway and the PI3K (phosphoinositide 3 kinase)/PKC pathway, both of which are involved in memory processing. These findings point to a convergence at the biochemical level between pathways involved in AD and those important for normal memory. PMID- 16246040 TI - Insulin and cognitive function in humans: experimental data and therapeutic considerations. AB - Data from experimental studies in animals and from epidemiological studies in humans suggest a link between insulin and cognitive performance. Do these results translate into clinical and therapeutic benefit for people with cognitive impairment? Insulin injected peripherally can readily cross the blood-brain barrier. Intravenous insulin can improve aspects of cognitive function in healthy adults and in individuals with Alzheimer's dementia. Moreover, intravenous insulin increases concentrations of a long form of beta-amyloid protein, Abeta42. One potential confounding factor with these data, however, is the need for co administration of glucose with the insulin to maintain euglycaemia as glucose itself can facilitate memory function. Administration of insulin via the intranasal route is scientifically (and therapeutically) more attractive because the insulin goes directly to the cerebrospinal fluid, with minimal systemic absorption; this obviates the need for a glucose infusion. Intranasal insulin may improve some aspects of memory in healthy individuals, but has yet to be studied in people with cognitive impairment. TZDs (thiazolidinediones) reduce peripheral insulin concentrations by enhancing insulin sensitivity. In adults with Type II (non-insulin-dependent) diabetes, TZD therapy improves memory function, but so does sulphonylurea therapy (which elevates peripheral insulin concentrations). Improved memory is linked to lower blood glucose concentrations, rather than altered insulin levels. However, major trials are currently under way examining the impact of TZDs in people with dementia. PMID- 16246041 TI - Increased risk of Alzheimer's disease in Type II diabetes: insulin resistance of the brain or insulin-induced amyloid pathology? AB - Type II diabetes mellitus (DM2) is associated with an increased risk of cognitive dysfunction and dementia. The increased risk of dementia concerns both Alzheimer's disease and vascular dementia. Although some uncertainty remains into the exact pathogenesis, several mechanisms through which DM2 may affect the brain have now been identified. First, factors related to the 'metabolic syndrome', a cluster of metabolic and vascular risk factors (e.g. dyslipidaemia and hypertension) that is closely linked to DM2, may be involved. A number of these risk factors are predictors of cerebrovascular disease, accelerated cognitive decline and dementia. Secondly, hyperglycaemia may be involved, through adverse effects of potentially 'toxic' glucose metabolites on the brain and its vasculature. Thirdly, insulin itself may be involved. Insulin can directly modulate synaptic plasticity and learning and memory, and disturbances in insulin signalling pathways in the periphery and in the brain have recently been implicated in Alzheimer's disease and brain aging. Insulin also regulates the metabolism of beta-amyloid and tau, the building blocks of amyloid plaques and neurofibrillary tangles, the neuropathological hallmarks of Alzheimer's disease. In this paper, the evidence for the association between DM2 and dementia and for each of these underlying mechanisms will be reviewed, with emphasis on the role of insulin itself. PMID- 16246042 TI - Adipose tissue function in the insulin-resistance syndrome. AB - Insulin resistance is often seen as a consequence of obesity and there are several possible links between adipose tissue function and insulin resistance determined in other organs such as skeletal muscle or liver. One such link is the regulation of NEFA (non-esterified fatty acid) delivery to the rest of the body. Simplistically, an expanded adipose tissue mass delivers more NEFA to the systemic circulation and these fatty acids compete for substrate utilization in skeletal muscle, which in turn reduces glucose utilization. This increases blood glucose concentration and provides the stimulus for increased insulin secretion and hyperinsulinaemia is a key feature of the insulin-resistance syndrome. However, there is abundant evidence that adipose tissue is exquisitely insulin sensitive and hyperinsulinaemia may therefore lead to a constant lipolytic inhibition in adipose tissue. Consequently, the main function of adipose tissue, to rapidly switch between fat uptake and fat release, will be hampered. Adipose tissue blood flow is the conveyor of signals and substrates to and from the adipose tissue. In healthy people adipose tissue blood flow is much enhanced by food intake, whereas in insulin-resistant subjects this response is blunted. This is another facet of unresponsiveness of adipose tissue in the insulin-resistance syndrome. PMID- 16246043 TI - Adipose tissue changes in obesity. AB - This review gives a broad description of some of the changes in adipose tissue seen in obesity. There are multiple changes in adipose tissue in obesity: histological, neural and vascular, relating to lipid and carbohydrate metabolism and to adipose tissue's endocrine functions. Some may originate from a simple physical expansion of cell size and number. It is unclear which are the most important either in terms of intermediary metabolism or of contributing to the co morbidities of obesity. Important questions for the future include the reversibility of obesity-related changes and indeed whether the changes differ between depots and species. Recent studies examining physiological regulation within adipose tissue demonstrate it to be relatively unresponsive to changes in everyday life. PMID- 16246045 TI - Regulation of lipid metabolism via angiopoietin-like proteins. AB - Regulation of mammalian energy metabolism is an intricate process involving numerous hormones, transcription factors and signal transduction cascades. Much of the regulation occurs via secreted factors that relay information from one organ to another. One group of secreted factors that recently emerged as having a major impact on lipid and possibly glucose metabolism are the ANGPTLs (angiopoietin-like proteins). This includes ANGPTL3, ANGPTL4/FIAF (fasting induced adipose factor), and ANGPTL6/AGF (angiopoietin-related growth factor). Although the receptors for these proteins have yet to be identified, it is nevertheless increasingly clear that these proteins have important effects on plasma triacylglycerol clearance, adipose tissue lipolysis, and adiposity. This review summarizes contemporary data on ANGPTLs with emphasis on the connection with energy metabolism. PMID- 16246044 TI - Mouse models of PPAR-gamma deficiency: dissecting PPAR-gamma's role in metabolic homoeostasis. AB - The identification of humans with mutations in PPAR-gamma (peroxisome proliferator-activated receptor-gamma) has underlined its importance in the pathogenesis of the metabolic syndrome. Genetically modified mice provide powerful tools to dissect the mechanisms by which PPAR-gamma regulates metabolic processes. Ablation of PPAR-gamma in vivo is lethal and thus dissection of PPAR gamma function using mouse models has relied on the development of tissue and isoform-specific ablation and mouse models of human mutations. These models exhibit phenotypes of partial PPAR-gamma impairment and are useful to elucidate how PPAR-gamma regulates specific metabolic processes. These murine models have confirmed the involvement of PPAR-gamma in adipose tissue development, maintenance and distribution. The mechanism involved in PPAR-gamma regulation of glucose homoeostasis is obscure as both agonism and partial impairment of PPAR gamma increase insulin sensitivity. While adipose tissue is likely to be the primary target for the insulin-sensitizing effects of PPAR-gamma, some murine models suggest PPAR-gamma expressed outside adipose tissue may also contribute actively to maintain glucose homoeostasis. Interestingly, mutations in PPAR-gamma that cause severe insulin resistance in humans when expressed in mice do not result in insulin insensitivity. However, these murine models can recapitulate the effects in fuel partitioning, post-prandial lipid handling and vasculature dysfunction observed in humans. In summary, these murine models of PPAR-gamma have provided useful in vivo systems to dissect the function of PPAR-gamma, but additionally have revealed a picture of complexity. These models have confirmed a key role for PPAR-gamma in the metabolic syndrome; however, they challenge the concept that insulin resistance is the main factor linking the clinical manifestations of the metabolic syndrome. PMID- 16246046 TI - Role of malonyl-CoA in the hypothalamic control of food intake and energy expenditure. AB - The brain plays an important role in the regulation of energy balance in higher animals. Global energy balance is monitored by sets of neurons in the hypothalamus that respond to peripheral hormonal and afferent neural signals that sense the energy status. Malonyl-CoA, an intermediate in the biosynthesis of fatty acids, appears to function in this hypothalamic energy-sensing system. The steady-state level of malonyl-CoA is determined by its rate of synthesis catalysed by ACC (acetyl-CoA carboxylase) relative to its rate of turnover catalysed by FAS (fatty acid synthase). Changes in the level of malonyl-CoA in the hypothalamus alter the expression/secretion of key hypothalamic orexigenic and anorexigenic neuropeptides that regulate the feeding behaviour and energy expenditure. Inhibitors of FAS, administered i.c.v. (intracerebroventricularly) to lean or obese mice, cause a rapid rise in hypothalamic malonyl-CoA level, suppression of food intake, increased fatty acid oxidation in skeletal muscle and profound weight loss. Stereotactic delivery of a viral MCD (malonyl-CoA decarboxylase) expression vector into the ventral hypothalamus lowers malonyl-CoA levels and reverses the anorectic effect of the FAS inhibitors. Fasting decreases, whereas refeeding increases, hypothalamic malonyl-CoA and alters subsequent feeding behaviour accordingly. The level of malonyl-CoA in the hypothalamus appears to be under the control of 5'-AMP kinase, which phosphorylates and thereby inactivates ACC under conditions of energy surplus. Thus malonyl-CoA appears to link the energy-responsive fatty acid synthesis in the hypothalamus to feeding behaviour and peripheral energy expenditure. PMID- 16246047 TI - Diet-induced obesity in the Sprague-Dawley rat: dietary manipulations and their effect on hypothalamic neuropeptide energy balance systems. AB - The SD (Sprague-Dawley) rat model of DIO (diet-induced obesity) is reported to exhibit a clear segregation into susceptible and resistant subpopulations shortly after transfer to a HE (high energy) diet. This does not appear to be the case for rats sourced in the U.K., where body weight gain on obesogenic HE diet is normally distributed, as might be anticipated for a polygenic trait in an outbred population. Many of the energy balance effects of dietary manipulation in this model (e.g. supplementation of HE diet with the liquid diet, Ensure; energy intake and defence of body weight following withdrawal of obesogenic diet) appear to be characteristics of the diets being manipulated rather than subject traits. The activities of energy balance-related hypothalamic signals are affected by diet and the development of DIO, but may not be able to differentiate between different diets and the relative levels of obesity that develop. PMID- 16246048 TI - Diseases of adipose tissue: genetic and acquired lipodystrophies. AB - Human lipodystrophies represent a group of diseases characterized by altered body fat amount and/or repartition and major metabolic alterations with insulin resistance leading to diabetic complications and increased cardiovascular and hepatic risk. Genetic forms of lipodystrophies are rare. Congenital generalized lipodystrophy or Berardinelli-Seip syndrome, autosomal recessive, is characterized by a complete early lipoatrophy and severe insulin resistance and results, in most cases, from mutations either in the seipin gene of unknown function or AGPAT2 encoding an enzyme involved in triacylglycerol synthesis. The Dunnigan syndrome [FPLD2 (familial partial lipodystrophy of the Dunnigan type)] is due to mutations in LMNA encoding the lamin A/C, belonging to the complex group of laminopathies that could comprise muscular and cardiac dystrophies, neuropathies and syndromes of premature aging. Some FPLDs are linked to loss-of function mutations in the PPAR-gamma gene (peroxisome-proliferator-activated receptor gamma; FPLD3) with severe metabolic alterations but a less severe lipodystrophy compared with FPLD2. The metabolic syndrome, acquired, represents the most common form of lipodystrophy. HIV-infected patients often present lipodystrophies, mainly related to side effects of antiretroviral drugs together with insulin resistance and metabolic alterations. Such syndromes help to understand the mechanisms involved in insulin resistance resulting from altered fat repartition and could benefit from insulin-sensitizing effects of lifestyle modifications or of specific medications. PMID- 16246049 TI - Signalling role of adipose tissue: adipokines and inflammation in obesity. AB - White adipose tissue (WAT) is a major endocrine and secretory organ, which releases a wide range of protein signals and factors termed adipokines. A number of adipokines, including leptin, adiponectin, tumour necrosis factor alpha, IL 1beta (interleukin 1beta), IL-6, monocyte chemotactic protein-1, macrophage migration inhibitory factor, nerve growth factor, vascular endothelial growth factor, plasminogen activator inhibitor 1 and haptoglobin, are linked to inflammation and the inflammatory response. Obesity is characterized by a state of chronic mild inflammation, with raised circulating levels of inflammatory markers and the expression and release of inflammation-related adipokines generally rises as adipose tissue expands (adiponectin, which has anti inflammatory action is an exception). The elevated production of inflammation related adipokines is increasingly considered to be important in the development of diseases linked to obesity, particularly Type II diabetes and the metabolic syndrome. WAT is involved in extensive cross-talk with other organs and multiple metabolic systems through the various adipokines. PMID- 16246050 TI - Protein aggregation, metals and oxidative stress in neurodegenerative diseases. AB - There is clear evidence implicating oxidative stress in the pathology of many different neurodegenerative diseases. ROS (reactive oxygen species) are the primary mediators of oxidative stress and many of the aggregating proteins and peptides associated with neurodegenerative disease can generate hydrogen peroxide, a key ROS, apparently through interactions with redox-active metal ions. Our recent results suggest that ROS are generated during the very early stages of protein aggregation, when protofibrils or soluble oligomers are present, but in the absence of mature amyloid fibrils. The generation of ROS during early-stage protein aggregation may be a common, fundamental molecular mechanism underlying the pathogenesis of oxidative damage, neurodegeneration and cell death in several different neurodegenerative diseases. Drugs that specifically target this process could be useful in the future therapy of these diseases. PMID- 16246051 TI - The role of cell-derived oligomers of Abeta in Alzheimer's disease and avenues for therapeutic intervention. AB - Burgeoning evidence suggests that soluble oligomers of Abeta (amyloid beta protein) are the earliest effectors of synaptic compromise in Alzheimer's disease. Whereas most other investigators have employed synthetic Abeta peptides, we have taken advantage of a beta-amyloid precursor protein-overexpressing cell line (referred to as 7PA2) that secretes sub-nanomolar levels of low-n oligomers of Abeta. These are composed of heterogeneous Abeta peptides that migrate on SDS/PAGE as dimers, trimers and tetramers. When injected into the lateral ventricle of rats in vivo, these soluble oligomers inhibit hippocampal long-term potentiation and alter the memory of a complex learned behaviour. Biochemical manipulation of 7PA2 medium including immunodepletion with Abeta-specific antibodies and fractionation by size-exclusion chromatography allowed us to unambiguously attribute these effects to low-n oligomers. Using this paradigm we have tested compounds directed at three prominent amyloid-based therapeutic targets: inhibition of the secretases responsible for Abeta production, inhibition of Abeta aggregation and immunization against Abeta. In each case, compounds capable of reducing oligomer production or antibodies that avidly bind Abeta oligomers also ameliorate the synaptotoxic effects of these natural, cell derived oligomers. PMID- 16246052 TI - Protein lipidation of BACE. AB - Our research has concentrated upon the protein lipid modification of BACE [beta site amyloid precursor protein cleaving enzyme (beta-secretase)], of which very little is currently known. Lipidation influences the production of Abeta (amyloid beta-protein) by promoting the dimerization of BACE. PMID- 16246053 TI - Glycosylation and misfolding of PrP. AB - The TSEs (transmissible spongiform encephalopathies) are not only devastating neurological diseases but also provide a biochemical conundrum; how can a disease agent replicate in the apparent absence of genetic material? The prion hypothesis proposes that the TSE agent is a misfolded form of the host glycoprotein PrP (prion protein). However, a number of questions regarding the hypothesis remain to be addressed. We are using gene-targeted PrP transgenics models to investigate these issues. Here we discuss our recent results that examine the importance of PrP's N-glycans to the misfolding of the protein. PMID- 16246054 TI - Expression and activity of beta-site amyloid precursor protein cleaving enzyme in Alzheimer's disease. AB - Several lines of evidence indicate that the Abeta peptide is involved at some level in the pathological process that results in the clinical symptoms of AD (Alzheimer's disease). The N-terminus of Abeta is generated by cleavage of the Met-Asp bond at position 671-672 of APP (amyloid precursor protein), catalysed by a proteolytic activity called beta-secretase. Two 'beta-secretase' proteases have been identified: BACE (beta-site APP-cleaving enzyme) and BACE2. The cause of sporadic AD is currently unknown, but some studies have reported elevated BACE/beta-secretase activity in brain regions affected by the disease. We have demonstrated that robust beta-secretase activity is also detectable in platelets that contain APP and release Abeta. This review considers the current evidence for alterations in beta-secretase activity, and/or alterations in BACE expression, in post-mortem brain tissue and platelets from individuals with AD. PMID- 16246056 TI - Alpha-synuclein aggregation in neurodegenerative diseases and its inhibition as a potential therapeutic strategy. AB - There is strong evidence for the involvement of alpha-synuclein in the pathologies of several neurodegenerative disorders, including PD (Parkinson's disease). Development of disease appears to be linked to processes that increase the rate at which alpha-synuclein forms aggregates. These processes include increased protein concentration (via either increased rate of synthesis or decreased rate of degradation), and altered forms of alpha-synuclein (such as truncations, missense mutations, or chemical modifications by oxidative reactions). Aggregated forms of the protein are toxic to cells and one therapeutic strategy would be to reduce the rate at which aggregation occurs. To this end we have designed several peptides that reduce alpha-synuclein aggregation. A cell-permeable version of one such peptide was able to inhibit the DNA damage induced by Fe(II) in neuronal cells transfected with alpha-synuclein (A53T), a familial PD-associated mutation. PMID- 16246055 TI - Abeta-degrading enzymes: modulators of Alzheimer's disease pathogenesis and targets for therapeutic intervention. AB - The accumulation of Abeta (amyloid beta-protein) peptides in the brain is a pathological hallmark of all forms of AD (Alzheimer's disease) and reducing Abeta levels can prevent or reverse cognitive deficits in mouse models of the disease. Abeta is produced continuously and its concentration is determined in part by the activities of several degradative enzymes, including NEP (neprilysin), IDE (insulin-degrading enzyme), ECE-1 (endothelin-converting enzyme 1) and ECE-2, and probably plasmin. Decreased activity of any of these enzymes due to genetic mutation, or age- or disease-related alterations in gene expression or proteolytic activity, may increase the risk for AD. Conversely, increased expression of these enzymes may confer a protective effect. Increasing Abeta degradation through gene therapy, transcriptional activation or even pharmacological activation of the Abeta-degrading enzymes represents a novel therapeutic strategy for the treatment of AD that is currently being evaluated in cell-culture and animal models. In this paper, we will review the roles of NEP, IDE, ECE and plasmin in determining endogenous Abeta concentration, highlighting recent results concerning the regulation of these enzymes and their potential as therapeutic targets. PMID- 16246057 TI - Structure and neurotoxicity of novel amyloids derived from the BRI gene. AB - A number of human neurodegenerative diseases involve aggregated amyloid proteins in the brain, e.g. Alzheimer's disease (beta-amyloid) and Parkinson's disease (alpha-synuclein). Other examples are rare familial dementias which involve the BRI gene. In a British family, mutation of the termination codon extends the reading frame of BRI to yield a furin-processed 34-residue peptide (Abri; British dementia peptide), 11 residues longer than the wild-type (WT). In a Danish family, a ten-base insertion also yields a 34-residue peptide (Adan; Danish dementia peptide). To explore the roles of Abri and Adan in neurodegeneration, we synthesized Abri and Adan in oxidized and reduced forms and generated transgenic mice colonies expressing the WT and mutated forms of BRI. We have generated transgenic mice colonies bearing the genes coding for WT-BRI, Adan and Abri under the control of the Thy1 promoter. Whereas WT-BRI transgenic mice express full length WT-BRI protein in their brains, Adan protein is fully processed to small peptides. PMID- 16246058 TI - Studies of the aggregation of an amyloidogenic alpha-synuclein peptide fragment. AB - The deposition of alpha-syn (alpha-synuclein) fibrils in Lewy bodies is a characteristic feature of individuals with neurodegenerative disorders. A peptide comprising the central residues 71-82 of alpha-syn [alpha-syn(71-82)] is capable of forming beta-sheet-rich, amyloid-like fibrils with similar morphologies to fibrils of the full-length protein, providing a useful model of pathogenic alpha syn fibrils that is suitable for detailed structural analysis. We have studied the morphology and gross structural features of alpha-syn(71-82) fibrils formed under different conditions in order to obtain reliable conditions for producing fibrils for further structural investigations. The results indicate that the rate of aggregation and the morphology of the fibrils formed are sensitive to pH and temperature. PMID- 16246059 TI - Novel heparan sulphate analogues: inhibition of beta-secretase cleavage of amyloid precursor protein. AB - The role of HS (heparan sulphate) in the pathology of AD (Alzheimer's disease) is multifaceted. HS and other glycosaminoglycans have been widely reported to be associated with neuritic plaques. HS has also been shown to promote the aggregation of Abeta (amyloid beta-peptide), the proteinaceous component of neuritic plaques. Recently, we described a novel and contrasting role for HS in the pathology of AD: HS can inhibit the formation of Abeta, by directly interacting with the protease BACE1 (beta-site amyloid precursor protein cleaving enzyme 1; beta-secretase 1), that cleaves the amyloid precursor protein and is the rate limiting step in the generation of Abeta. Here, we review the current roles of HS and the potential for HS-derivatives in the treatment of AD. PMID- 16246060 TI - Negative feedback in NO/cGMP signalling. AB - Most of the effects of the signalling molecule nitric oxide (NO) are mediated by the stimulation of the NO-sensitive GC (guanylate cyclase) and the subsequent increase in cGMP formation. The enzyme contains a prosthetic haem group, which mediates NO stimulation. In addition to the physiological activator NO, NO sensitizers like the substance YC-1 sensitize the enzyme towards NO and may therefore have important pharmacological implications. Two isoforms of NO sensitive GC have been identified to date that share regulatory properties, but differ in the subcellular localization. The more ubiquitously expressed alpha1beta1 heterodimer and the alpha2beta1 isoform are mainly expressed in brain. In intact cells, NO-induced cGMP signalling not only depends on cGMP formation, but is also critically determined by the activity of the enzymes responsible for cGMP degradation, e.g. PDE5 (phosphodiesterase 5). Recently, direct activation of PDE5 by cGMP was demonstrated, limiting the cGMP increase and thus functioning as a negative feedback. As the cGMP-induced PDE5 activation turned out to be sustained, in the range of hours, it is probably responsible for the NO-induced desensitization observed within NO/cGMP signalling. PMID- 16246061 TI - Reactive oxygen species (ROS)-mediated beta-cleavage of the prion protein in the mechanism of the cellular response to oxidative stress. AB - The PrP(C) [cellular isoform of PrP (prion protein)] can undergo a conformational conversion to produce a proteinase-resistant form PrP(Sc) (scrapie isoform of PrP), a step critical for the development of prion disease. Although essential for disease progression, the normal cellular function of PrP(C) remains unknown. Suggestions to date have centred on a protective role against oxidative stress. We have demonstrated that ROS (reactive oxygen species)-mediated beta-cleavage of PrP(C) occurs at the cell surface, can be inhibited following hydroxyl radical quenching and has a prerequisite for the octarepeat region in the N-terminus of the protein. Significantly, two disease-associated mutants of PrP, namely PG14 and A116V (Ala(116)-->Val), were unable to undergo beta-cleavage and this lack of proteolysis was accompanied by functional consequences in cells expressing these mutant proteins. The cells were found to be less viable following exposure to copper and H2O2, had reduced levels of glutathione peroxidase and increased amounts of intracellular oxygen radicals. These results suggest that beta cleavage of PrP(C) is an initial consequence following exposure to ROS in the extracellular environment contributing to a pathway involved in antioxidant protection of neuronal cells. PMID- 16246062 TI - Inhibition of pro-inflammatory cytokine receptor signalling by cAMP in vascular endothelial cells. AB - The anti-inflammatory effects of the prototypical second messenger cAMP have been extensively documented in multiple cell types. However, in many instances, the molecular mechanisms by which cAMP elevation disrupts specific pro-inflammatory signalling cascades are unknown. In this review, we will describe the importance of the JAK-STAT (where JAK stands for Janus kinase and STAT for signal transducer and activator of transcription) signalling pathway in vascular endothelial cell function, outline key inhibitory processes that serve to reduce cytokine stimulated tyrosine phosphorylation and activation of STAT proteins, and discuss possible mechanisms by which intracellular cAMP sensors could interact with these inhibitory processes to diminish cytokine receptor-mediated pro-inflammatory signalling. PMID- 16246063 TI - Novel tissue remodelling roles for human recombinant erythropoietin. AB - rHuEPO (recombinant human erythropoietin) is a haemopoietic growth factor and a primary regulator of erythropoiesis that is used for the treatment of chronic anaemia associated with RA (rheumatoid arthritis). Erythropoietin also appears to modulate a broad array of cellular processes, including progenitor stem-cell development, cellular integrity, angiogenesis and oxidative damage. These diverse activities suggest the exciting possibility of multiple roles for rHuEPO therapy in a variety of disorders other than RA, including cerebral ischaemia, myocardial infarction, chronic congestive heart failure and cancer. Thus it appears that rHuEPO may be a pleiotropic agent, capable of influencing tissue remodelling independently of its established erythropoietic role. Whereas these effects may be largely beneficial, dose-related side effects could have implications for the safe therapeutic use of rHuEPO and its illegal use as a performance-enhancing agent in endurance sports. PMID- 16246064 TI - Compartmentation of G-protein-coupled receptors and their signalling components in lipid rafts and caveolae. AB - G-protein-coupled receptors (GPCRs) and post-GPCR signalling components are expressed at low overall abundance in plasma membranes, yet they evoke rapid, high-fidelity responses. Considerable evidence suggests that GPCR signalling components are organized together in membrane microdomains, in particular lipid rafts, enriched in cholesterol and sphingolipids, and caveolae, a subset of lipid rafts that also possess the protein caveolin, whose scaffolding domain may serve as an anchor for signalling components. Caveolae were originally identified based on their morphological appearance but their role in compartmentation of GPCR signalling has been primarily studied by biochemical techniques, such as subcellular fractionation and immunoprecipitation. Our recent studies obtained using both microscopic and biochemical methods with adult cardiac myocytes show expression of caveolin not only in surface sarcolemmal domains but also at, or close to, internal regions located at transverse tubules/sarcoplasmic reticulum. Other results show co-localization in lipid rafts/caveolae of AC (adenylyl cyclase), in particular AC6, certain GPCRs, G-proteins and eNOS (endothelial nitric oxide synthase; NOS3), which generates NO, a modulator of AC6. Existence of multiple caveolin-rich microdomains and their expression of multiple modulators of signalling strengthen the evidence that caveolins and lipid rafts/caveolae organize and regulate GPCR signal transduction in eukaryotic cells. PMID- 16246065 TI - Expression cloning of novel regulators of 92 kDa type IV collagenase expression. AB - Overexpression of the 92 kDa type IV collagenase (MMP-9) contributes to cancer progression. However, to date, there are few known regulators of expression of this metalloproteinase. We employed an expression library comprising 500,000 cDNA clones to screen for novel regulators of MMP-9 expression. HT1080 cells were transiently co-transfected with an MMP-9 promoter-luciferase reporter and pools of the cDNA expression library. Positive-scoring pools were subdivided in secondary and tertiary screens, after which the regulatory cDNAs were identified by DNA sequencing. This brief review illustrates the utility of expression cloning in identifying specific regulators of MMP-9 expression. PMID- 16246066 TI - Atypical L-type channels are down-regulated in hypoxia. AB - One type of cellular response to hypoxia is an increase in cytosolic Ca2+. VDCCs (voltage-dependent calcium channels) open upon membrane depolarization allowing inward current of Ca2+ ions. Two of the so-called L-type VDCC alpha1 subunits, Ca(v)1.2 and Ca(v)1.3, are found in the brain. We sought to investigate the effect of chronic hypoxia or treatment with a hypoxia-mimicking agent DFX (desferrioxamine mesylate) on expression of L-type VDCC in the SH-SY5Y neuroblastoma cell line. Western blotting identified two atypical forms of the L type channel with apparent molecular masses of approx. 100 and 150 kDa, compared with typical forms of approx. 200 kDa. Immunofluorescence microscopy shows the approx. 100 kDa protein located within the cell and on the cell surface, while the approx. 150 kDa protein is intracellular with punctate staining. Further analysis revealed that this approx. 150 kDa protein co-localizes with nuclear proteins but not with markers for other intracellular compartments. In addition, these proteins are both down-regulated in DFX-treated and hypoxic cells, suggesting that the mechanism of down-regulation is along the HIF (hypoxia inducible factor) pathway. This atypical localization of the 150 kDa protein suggests that it might play a role in nuclear calcium signalling in health and disease. PMID- 16246067 TI - Protein and lipid motifs regulate phosphatidylserine traffic in yeast. AB - Phosphatidylserine (PtdSer) is synthesized in the endoplasmic reticulum and its subdomains associated with the mitochondria [MAM (mitochondria-associated membrane)] and subsequently transported to the loci of the PtdSer decarboxylases, Pds1p (phosphatidylserine decarboxylase 1 encoded by the PSD1 gene that complements psd1 mutations) in the mitochondria, and Psd2p (PtdSer decarboxylase 2 encoded by the PSD2 gene that complements psd2 mutations) in the Golgi. Decarboxylation of PtdSer to PtdEtn (phosphatidylethanolamine) can be used as a biochemical indicator of transport to these organelles, which is regulated by specific lipid and protein motifs. PtdSer transport to mitochondria is controlled by ubiquitination via the action of the ubiquitin ligase subunit Met30p (a ubiquitin ligase subunit encoded by the MET30 gene that complements the met30 mutation affecting methionine biosynthesis). Mutant strains with lesions in the MET30 gene are defective in PtdSer transport and show altered ubiquitination of specific target proteins, such as the transcription factor Met4p (a transcription factor encoded by the MET4 gene that complements the met4 mutation affecting methionine biosynthesis). Mutations to MET30 cause defects in both the MAM as a donor of PtdSer, and the mitochondria as an acceptor of PtdSer in the transport reaction. PtdSer transport to the locus of Psd2p is controlled by specific protein and lipid motifs. The C2 (Ca2+ and phospholipid-binding sequence) domain of Psd2p, and the lipid-binding protein PstB2p (PtdSer transport B pathway protein encoded by the PSTB2 gene that complements the pstB2 mutation affecting PtdSer transport), must be present on acceptor membranes for PtdSer transport to occur. In addition, the action of the PtdIns 4-kinase, Stt4p (PtdIns 4-kinase encoded by the STT4 gene that complements the stt4 mutation causing staurosporine and temperature-sensitive growth) is also required for PtdSer transport to the locus of Psd2p. Reconstitution of PtdSer transport to Psd2p using liposomes demonstrates that PtdSer-rich domains present in vesicles are preferred substrates for transport. In addition, the incorporation of phosphatidic acid into donor membranes enhances the rate of PtdSer transport. Collectively, these data support a model for PtdSer transport in which specific proteins and lipids are required on donor and acceptor membranes. PMID- 16246068 TI - The contributions of biosynthesis and acyl chain remodelling to the molecular species profile of phosphatidylcholine in yeast. AB - Phosphatidylcholine (PC) is a very abundant membrane lipid in most eukaryotes, including yeast. The molecular species profile of PC, i.e. the ensemble of PC molecules with acyl chains differing in number of carbon atoms and double bonds, is important for membrane function. Pathways of PC synthesis and turnover maintain PC homoeostasis and determine the molecular species profile of PC. Studies addressing the processes involved in establishing the molecular species composition of PC in yeast using stable isotope labelling combined with detection by MS are reviewed. PMID- 16246069 TI - Regulation of phospholipid synthesis in yeast by zinc. AB - The yeast Saccharomyces cerevisiae has the ability to cope with a variety of stress conditions (e.g. zinc deficiency) by regulating the expression of enzyme activities including those involved with phospholipid synthesis. Zinc is an essential mineral required for the growth and metabolism of S. cerevisiae. Depletion of zinc from the growth medium of wild-type cells results in alterations in phospholipid composition including an increase in PI (phosphatidylinositol) and a decrease in phosphatidylethanolamine. These changes can be attributed to an increase in PIS1-encoded PI synthase activity and a decrease in the activities of several CDP-diacylglycerol pathway enzymes including the CHO1-encoded PS (phosphatidylserine) synthase. The reduction in PS synthase in response to zinc depletion is due to a repression mechanism that involves the UAS(INO) (inositol upstream activating sequence) element in the CHO1 promoter and the negative transcription factor Opi1p. These factors are also responsible for the inositol-mediated repression of CHO1. This regulation may play an important role in allowing cells to adapt to zinc deficiency given the essential roles that phospholipids play in the structure and function of cellular membranes. PMID- 16246070 TI - In Saccharomyces cerevisiae, impaired PRPP synthesis is accompanied by valproate and Li+ sensitivity. AB - The biosynthetic intermediate PRPP (phosphoribosylpyrophosphate) has a central role in cellular biochemistry since it links carbon and nitrogen metabolism. Its importance may be reflected in the fact that, in the Saccharomyces cerevisiae (yeast) genome, there are five unlinked genes, PRS1-PRS5, each of which is theoretically capable of encoding the enzyme synthesizing PRPP. Interference with the complement of PRS genes in S. cerevisiae has far-reaching consequences for yeast physiology and has uncovered unexpected metabolic links including cell wall integrity and phospholipid metabolism. PMID- 16246071 TI - Deficiency in mitochondrial anionic phospholipid synthesis impairs cell wall biogenesis. AB - Cardiolipin (CL) is the signature lipid of the mitochondrial membrane and plays a key role in mitochondrial physiology and cell viability. The importance of CL is underscored by the finding that the severe genetic disorder Barth syndrome results from defective CL composition and acylation. Disruption of PGS1, which encodes the enzyme that catalyses the committed step of CL synthesis, results in loss of the mitochondrial anionic phospholipids phosphatidylglycerol and CL. The pgs1Delta mutant exhibits severe growth defects at 37 degrees C. To understand the essential functions of mitochondrial anionic lipids at elevated temperatures, we isolated suppressors of pgs1Delta that grew at 37 degrees C. The present review summarizes our analysis of suppression of pgs1Delta growth defects by a mutant that has a loss-of-function mutation in KRE5, a gene involved in cell wall biogenesis. PMID- 16246072 TI - Mitochondrial fatty acid synthesis and maintenance of respiratory competent mitochondria in yeast. AB - Mitochondrial FAS (fatty acid synthesis) of type II is a widely conserved process in eukaryotic organisms, with particular importance for respiratory competence and mitochondrial morphology maintenance in Saccharomyces cerevisiae. The recent characterization of three missing enzymes completes the pathway. Etr1p (enoyl thioester reductase) was identified via purification of the protein followed by molecular cloning. To study the link between FAS and cell respiration further, we also created a yeast strain that has FabI enoyl-ACP (acyl-carrier protein) reductase gene from Escherichia coli engineered to carry a mitochondrial targeting sequence in the genome, replacing the endogenous ETR1 gene. This strain is respiratory competent, but unlike the ETR1 wild-type strain, it is sensitive to triclosan on media containing only non-fermentable carbon source. A colony colour-sectoring screen was applied for cloning of YHR067w/RMD12, the gene encoding mitochondrial 3-hydroxyacyl-ACP dehydratase (Htd2/Yhr067p), the last missing component of the mitochondrial FAS. Finally, Hfa1p was shown to be the mitochondrial acetyl-CoA carboxylase. PMID- 16246073 TI - Using genomic and lipidomic strategies to investigate sphingolipid function in the yeast heat-stress response. AB - In addition to their crucial role in membrane structure in Saccharomyces cerevisiae, sphingolipids serve vital roles in various aspects of yeast biology including endocytosis, intracellular protein transport and stress responses. Although previous studies have unequivocally demonstrated the sphingolipid requirements for these processes, few studies have contributed mechanistic information. We have used a systems approach including microarray, lipidomics and metabolic modelling to better understand (i) biochemical relationships between various branches of sphingolipid metabolism and pathways and contributing pathways such as fatty acid metabolism and phospholipid synthesis, (ii) the changes in cellular sphingolipid composition under various conditions and (iii) the effects of these changes on the transcriptional profiles and subsequently, cell phenotypes. Thus far, these approaches have indicated roles for sphingolipids in major transcriptional changes in response to heat stress, carbon source utilization, sporulation, cell wall integrity and other basic cellular functions. Although the yeast genome is fully sequenced, nearly 50% of all transcribed open reading frames remain uncharacterized with regard to cellular function; therefore, a major advantage of this approach is the ability to identify both biochemical and biological roles for enzymes and their products within broad cellular contexts. PMID- 16246074 TI - Signalling functions for sphingolipid long-chain bases in Saccharomyces cerevisiae. AB - Over the past several years, studies of sphingolipid functions in the baker's yeast Saccharomyces cerevisiae have revealed that the sphingoid LCBs (long-chain bases), dihydrosphingosine and PHS (phytosphingosine), are important signalling molecules or second messengers under heat stress and during non-stressed conditions. LCBs are now recognized as regulators of AGC-type protein kinase (where AGC stands for protein kinases A, G and C) Pkh1 and Pkh2, which are homologues of mammalian phosphoinositide-dependent protein kinase 1. LCBs were previously shown to activate Pkh1 and Pkh2, which then activate the downstream protein kinase Pkc1. We have recently demonstrated that PHS stimulates Pkh1 to activate additional downstream kinases including Ypk1, Ypk2 and Sch9. We have also found that PHS acts downstream of Pkh1 and partially activates Ypk1, Ypk2 and Sch9. These kinases control a wide range of cellular processes including growth, cell wall integrity, stress resistance, endocytosis and aging. As we learn more about the cellular processes controlled by Ypk1, Ypk2 and Sch9, we will have a far greater appreciation of LCBs as second messengers. PMID- 16246075 TI - Formation and mobilization of neutral lipids in the yeast Saccharomyces cerevisiae. AB - Since energy storage is a basic metabolic process, the synthesis of neutral lipids occurs in all kingdoms of life. The yeast Saccharomyces cerevisiae, widely accepted as a model eukaryotic cell, contains two classes of neutral lipids, namely STEs (steryl esters) and TAGs (triacylglycerols). TAGs are synthesized through two pathways governed by the acyl-CoA diacylglycerol acyltransferase Dga1p and the phospholipid diacylglycerol acyltransferase Lro1p. STEs are formed by two STE synthases Are1p and Are2p, two enzymes with overlapping function, which also catalyse TAG formation, although to a minor extent. Neutral lipids are stored in the so-called lipid particles and can be utilized for membrane formation under conditions of lipid depletion. For this purpose, storage lipids have to be mobilized by TAG lipases and STE hydrolases. A TAG lipase named Tgl3p was identified as a major yeast TAG hydrolytic enzyme in lipid particles. Recently, a new family of hydrolases was detected which is required for STE mobilization in S. cerevisiae. These enzymes, named Yeh1p, Yeh2p and Tgl1p, are paralogues of the mammalian acid lipase family. The role of these proteins in biosynthesis and mobilization of TAG and STE, and the regulation of these processes will be discussed in this minireview. PMID- 16246076 TI - Interactions of the ergosterol biosynthetic pathway with other lipid pathways. AB - Micro-organisms have recently received broad attention as sources of novel lipids. An increased understanding of the effects of fats and oils and their composition on the metabolism and on health has shifted the focus towards the use of lipids for disease treatment and prevention and for the promotion of good health. A large range of lipidic products produced by yeast is known today. Ergosterol and its metabolic precursors are major lipidic components of industrial and commercial interest. Having in mind the aim to increase the productivity of ergosterol and its precursor metabolites, both the knowledge of regulatory mechanisms of the biosynthetic pathway and its interactions with other lipid pathways like those of sphingolipids, phospholipids and fatty acids are crucial. PMID- 16246077 TI - Homoeostatic systems for sterols and other lipids. AB - Fatty acids and sterols are vital components of all eukaryotic cells. Both are used as building blocks for numerous cellular processes such as membrane biosynthesis or hormone production (sterols). Furthermore, these compounds elicit a variety of effects intracellularly as they can act as signalling molecules and regulate gene expression. The metabolism of fatty acids and sterols represents a very intricate network of pathways that are regulated in a precise manner in order to maintain lipid homoeostasis within a cell. Using the budding yeast Saccharomyces cerevisiae as a model system, we touch upon some of the aspects of achieving and maintaining this lipid homoeostasis. PMID- 16246078 TI - Saccharomyces cerevisiae, a model to study sterol uptake and transport in eukaryotes. AB - The molecular mechanisms that govern intracellular transport of sterols in eukaryotic cells are only poorly understood. Saccharomyces cerevisiae is a facultative anaerobic organism that requires supplementation with unsaturated fatty acids and sterols to grow in the absence of oxygen, as the synthesis of these lipids requires molecular oxygen. The fact that yeast grows well under anaerobic conditions indicates that lipid uptake is rapid and efficient. To identify components in this lipid uptake and transport pathway, we screened the yeast mutant collection for genes that are essential under anaerobic conditions. Out of the approx. 4800 non-essential genes represented in the mutant collection, 37 were required for growth under anaerobic conditions. Uptake assays using radiolabelled cholesterol revealed that 16 of these genes are required for cholesterol uptake/transport and esterification. Further characterization of the precise role of these genes is likely to advance our understanding of this elusive pathway in yeast and may prove to be relevant to understand sterol homoeostasis in higher eukaryotic cells. PMID- 16246079 TI - Enzyme redesign and interactions of substrate analogues with sterol methyltransferase to understand phytosterol diversity, reaction mechanism and the nature of the active site. AB - Several STM (sterol methyltransferase) genes have been cloned, sequenced and expressed in bacteria recently, making it possible to address questions of the relationship between sterol structure and function. The active site and mechanism of action of a set of phylogenetically diverse SMTs have been probed by site directed mutagenesis as well as by using substrate and related analogues of the SMT-catalysed reaction. An active-site model has been developed that is in accord with the results presented, which is consistent with the hypothesis that SMTs are bifunctional enzymes kinetically responsible to bind Delta24-acceptor sterols of specific steric and electronic character and rigid orientation imposed by multiple hydrophobic active site contacts exacted from a common waxy core. Functional divergence influenced by the architectural role of sterols in membranes is considered to govern the evolution of product distribution and specificity of individual SMTs as discussed. PMID- 16246080 TI - Single amino acid exchanges in FAD-binding domains of squalene epoxidase of Saccharomyces cerevisiae lead to either loss of functionality or terbinafine sensitivity. AB - Squalene epoxidase (Erg1p) is an essential enzyme in the ergosterol biosynthesis pathway in yeast. For its enzymatic activity, Erg1p requires molecular oxygen, NAD(P)H and FAD. Amino acid analysis and sequence alignment with other squalene epoxidases revealed two highly conserved FAD-binding domains, FAD I and FAD II. By random PCR mutagenesis of the ERG1 gene, one erg1 allele was isolated that carries a mutation leading to a single amino acid exchange in the FAD I domain close to the N-terminus of Erg1p. This erg1 allele codes for functional squalene epoxidase and renders yeast cells hypersensitive to terbinafine. Amino acid exchanges of other conserved residues in the FAD I and FAD II regions either led to non-functional squalene epoxidase or to the formation of squalene epoxidase with wild-type properties. These results describe the importance of specific amino acids for enzymatic activity in the yeast squalene epoxidase Erg1p. PMID- 16246081 TI - Access of the substrate to the active site of squalene and oxidosqualene cyclases: comparative inhibition, site-directed mutagenesis and homology modelling studies. AB - Substrate access to the active-site cavity of squalene-hopene cyclase from Alicyclobacillus acidocaldarious and lanosterol synthase [OSC (oxidosqualene cyclase)] from Saccharomyces cerevisiae was studied by an inhibition, mutagenesis and homology-modelling approach. Crystal structure and homology modelling indicate that both enzymes possess a narrow constriction that separates an entrance lipophilic channel from the active-site cavity. The role of the constriction as a mobile gate that permits substrate passage was investigated by experiments in which critically located Cys residues, either present in native protein or inserted by site-directed mutagenesis, were labelled with specifically designed thiol-reacting molecules. Some amino acid residues of the yeast enzyme, selected on the basis of sequence alignment and a homology model, were individually replaced by residues bearing side chains of different lengths, charges or hydrophobicities. In some of these mutants, substitution severely reduced enzymatic activity and thermal stability. Homology modelling revealed that in these mutants some critical stabilizing interactions could no longer occur. The possible critical role of entrance channel and constriction in specific substrate recognition by eukaryotic OSC is discussed. PMID- 16246082 TI - Two mutants selectively resistant to polyenes reveal distinct mechanisms of antifungal activity by nystatin and amphotericin B. AB - Polyene macrolides nystatin and amphotericin B are widely used in the treatment of fungal infections. In order to characterize factors affecting polyene activity, we have isolated Saccharomyces cerevisiae mutants showing selective resistance to nystatin and amphotericin B. Characterization of two of these mutants (nystatin-resistant mutant X1/16 and amphotericin B-resistant mutant X3/33) is presented. Genetic analysis revealed that resistance in each of these mutants is caused by a mutation in one gene with a different mode of inheritance. Nystatin resistance in mutant X1/16 is caused by changes in sterol spectrum while amphotericin B resistance in mutant X3/33 is probably related to modification of the cell wall. Our results suggest that, in spite of their structural similarity, nystatin and amphotericin B differ significantly in mechanisms of their antifungal activity. PMID- 16246083 TI - Azole susceptibility and resistance in Candida dubliniensis. AB - Candida dubliniensis is a recently described species of pathogenic yeast that shares many phenotypic features with Candida albicans. It is primarily associated with oral colonization and infection in HIV-infected individuals. Isolates of C. dubliniensis are generally susceptible to commonly used azole antifungal agents; however, resistance has been observed in clinical isolates and can be induced by in vitro exposure. Molecular mechanisms of azole resistance in C. dubliniensis include increased drug efflux, modifications of the target enzyme and alterations in the ergosterol biosynthetic pathway. PMID- 16246084 TI - Regulation of sterol metabolism in Candida albicans by the UPC2 gene. AB - Candida albicans is an important pathogenic fungus of humans, causing a range of infections. These infections are usually treated with antifungal drugs that target sterol metabolism. Resistance to these antifungals can result from overexpression of sterol biosynthetic genes. Therefore it is of interest to understand transcriptional regulation of sterol biosynthesis in C. albicans. Recently two reports [Silver, Oliver and White (2004) Eukaryot. Cell 3, 1391 1397; MacPherson, Akache, Weber, De Deken, Raymond and Turcotte (2005) Antimicrob. Agents Chemother. 49, 1745-1752] have identified and characterized a single C. albicans transcription factor gene UPC2 that regulates sterol metabolism. The details of both characterizations are compared and contrasted. These reports extend our understanding of sterol regulation in this important human pathogen. PMID- 16246085 TI - Membrane raft lipid constituents affect drug susceptibilities of Candida albicans. AB - By exploiting the biosynthetic pathways of raft lipid constituents, in this study we demonstrate that fluctuations in either sphingolipid or ergosterol levels result in increased drug sensitivity and morphological defects in Candida albicans cells. We show that any change in either ergosterol composition by conditionally disrupting ERG1 or in sphingolipid composition by homozygously disrupting its biosynthetic gene IPT1 leads to improper surface localization of a major ABC (ATP-binding cassette) drug efflux protein, Cdr1p. Results suggest that sterol/sphingolipid-rich membrane microdomains play an important role in positioning and functional maintenance of the integral efflux protein. The impaired ability of erg1/ipt1 mutant cells to efflux drugs mediated through Cdr1p appears to be the main cause of increased drug sensitivity of Candida cells. PMID- 16246086 TI - The structure of the bacterial protein translocation complex SecYEG. AB - Proteins destined for secretion, membrane insertion or organellar import contain signal sequences that direct them to the membrane. Once there, transport machines receive and translocate them appropriately across or into the membrane. The related SecY and Sec61 protein translocation complexes are ubiquitous components of machines that are essential for protein transport. They co-operate with various partners such that the substrate polypeptide is pulled or pushed through the membrane by post- or co-translational mechanisms. In bacteria and archaea, the SecY complex (SecYEG/SecYEbeta) is a heterotrimer, which associates with ribosomes so that the polypeptide is threaded through the channel during its synthesis. Bacteria possess an additional pathway, whereby the newly synthesized substrate protein is maintained in an unfolded conformation and is engaged by the ATPase SecA and delivered to the translocon. Recent medium- (cryo-electron microscopy) and high-resolution (X-ray) structures of the Sec complex have dramatically increased our understanding about how proteins pass through membranes, but have posed a number of new questions. The Sec complex is active as an oligomer, but the structure indicates that the protein-conducting channel is formed by a monomer of SecYEG. Structures of the membrane-bound dimer of Escherichia coli SecYEG and the detergent-solubilized monomer of Methanococcus jannaschii SecYEbeta will be described and discussed in the context of the mechanism that underlies protein secretion and membrane insertion. PMID- 16246087 TI - Alternative mechanisms of initiating translation of mammalian mRNAs. AB - Of all the steps in mRNA translation, initiation is the one that differs most radically between prokaryotes and eukaryotes. Not only is there no equivalent of the prokaryotic Shine-Dalgarno rRNA-mRNA interaction, but also what requires only three initiation factor proteins (aggregate size approximately 125 kDa) in eubacteria needs at least 28 different polypeptides (aggregate >1600 kDa) in mammalian cells, which is actually larger than the size of the 40 S ribosomal subunit. Translation of the overwhelming majority of mammalian mRNAs occurs by a scanning mechanism, in which the 40 S ribosomal subunit, primed for initiation by the binding of several initiation factors including the eIF2 (eukaryotic initiation factor 2)-GTP-MettRNA(i) complex, is loaded on the mRNA immediately downstream of the 5'-cap, and then scans the RNA in the 5'-->3' direction. On recognition of (usually) the first AUG triplet via base-pairing with the Met tRNA(i) anticodon, scanning ceases, triggering GTP hydrolysis and release of eIF2 GDP. Finally, ribosomal subunit joining and the release of the other initiation factors completes the initiation process. This sketchy outline conceals the fact that the exact mechanism of scanning and the precise roles of the initiation factors remain enigmatic. However, the factor requirements for initiation site selection on some viral IRESs (internal ribosome entry sites/segments) are simpler, and investigations into these IRES-dependent mechanisms (particularly picornavirus, hepatitis C virus and insect dicistrovirus IRESs) have significantly enhanced our understanding of the standard scanning mechanism. This article surveys the various alternative mechanisms of initiation site selection on mammalian (and other eukaryotic) cellular and viral mRNAs, starting from the simplest (in terms of initiation factor requirements) and working towards the most complex, which paradoxically happens to be the reverse order of their discovery. PMID- 16246088 TI - Regulation of actin assembly by SCAR/WAVE proteins. AB - Actin reorganization is a tightly regulated process that co-ordinates complex cellular events, such as cell migration, chemotaxis, phagocytosis and adhesion, but the molecular mechanisms that underlie these processes are not well understood. SCAR (suppressor of cAMP receptor)/WAVE [WASP (Wiskott-Aldrich syndrome protein)-family verprolin homology protein] proteins are members of the conserved WASP family of cytoskeletal regulators, which play a critical role in actin dynamics by triggering Arp2/3 (actin-related protein 2/3)-dependent actin nucleation. SCAR/WAVEs are thought to be regulated by a pentameric complex which also contains Abi (Abl-interactor), Nap (Nck-associated protein), PIR121 (p53 inducible mRNA 121) and HSPC300 (haematopoietic stem progenitor cell 300), but the structural organization of the complex and the contribution of its individual components to the regulation of SCAR/WAVE function remain unclear. Additional features of SCAR/WAVE regulation are highlighted by the discovery of other interactors and distinct complexes. It is likely that the combinatorial assembly of different components of SCAR/WAVE complexes will prove to be vital for their roles at the centre of dynamic actin reorganization. PMID- 16246089 TI - SH3 domain-containing proteins and the actin cytoskeleton in yeast. AB - SH3 (Src homology-3) domains are involved in protein-protein interactions through proline-rich domains. Many SH3-containing proteins are implicated in actin cytoskeleton organization. The aim of our ongoing work is to study the functions of the SH3-containing proteins in actin cytoskeleton regulation. The yeast Saccharomyces cerevisiae proteome includes 29 SH3 domains distributed in 25 proteins. We have examined the direct involvement of these SH3 domains in actin polymerization using an in vitro polymerization assay on GST (glutathione S transferase)-SH3-coated beads. As expected, not all SH3 domains show polymerization activity, and many recruit distinct partners as assessed by microscopy and pull-down experiments. One such partner, Las17p, the yeast homologue of WASP (Wiskott-Aldrich syndrome protein), was assayed because it stimulates actin nucleation via the Arp2/3 (actin-related protein 2/3) complex. Ultimately, proteins involved in specific biological processes, such as membrane trafficking, may also be recruited by some of these SH3 domains, shedding light on the SH3-containing proteins and actin cytoskeleton functions in these processes. PMID- 16246090 TI - Coupling the dynamics of two actin networks--new views on the mechanics of cell protrusion. AB - We study how mechanical forces integrate spatially and temporally with regulatory signals at the leading edge of migrating cells. To probe the dynamics of this system, we developed quantitative fluorescent speckle microscopy, which maps out actin cytoskeleton transport, assembly and disassembly with high spatial resolution. Statistical processing of single speckle properties revealed two kinetically, kinematically and molecularly distinct, yet spatially overlapping, actin arrays at the leading edge of migrating epithelial cells. The first network, referred to as the lamellipodium, polymerizes and depolymerizes 1-2 microm from the edge in an Arp2/3 (actin-related protein 2/3)- and cofilin dependent fashion. The second network, referred to as the lamella, exhibits Arp2/3-independent polymerization. To elucidate the dynamic relationship between the two networks, we have begun to examine how assembly and flow are temporally modulated with respect to a protrusion event. In control cells we found bursts of protrusion preceding bursts of F-actin assembly. The time lag disappears in cells where Arp2/3-function is impaired. This and other results allowed us to propose a model in which tropomyosin protects lamella filaments from branching and severing, and to conjecture that Arp2/3-mediated lamellipodium assembly is a natural consequence of lamella expansion, but not the initiator of cell protrusion. PMID- 16246091 TI - Dystroglycan: a multifunctional adaptor protein. AB - Dystroglycan, a ubiquitous membrane-spanning cell adhesion molecule, is a crucial link between the actin cytoskeleton and the extracellular matrix. With a wide expression pattern and multiple interacting proteins, not only is dystroglycan now thought to be important as a structural molecule but also new research has suggested that it has a role in cell signalling, cytoskeleton reorganization and as a potential tumour suppressor. PMID- 16246092 TI - Formins and VASPs may co-operate in the formation of filopodia. AB - Filopodia are finger-like cell protrusions composed of parallel arrays of actin filaments, which elongate through actin polymerization at their tips. These highly dynamic structures seem to be used by many cell types as sensing organs to explore environmental cues and have been implicated in cell motility as well as in cell-substrate adhesion. Formins are highly conserved multidomain proteins that play important roles in the nucleation of actin and the formation of linear actin filaments, yet their role in filopodia formation has remained poorly defined. The Dictyostelium diaphanous-related formin dDia2 is strongly enriched in filopodia tips. Genetic and biochemical analysis revealed that this protein is important for cell migration and cell adhesion, but most importantly for the formation of filopodia. Recently, we have identified the Dictyostelium VASP (vasodilator-stimulated phosphoprotein) orthologue as a binding partner of dDia2 and provide evidence for a co-operative role of both proteins in filopodia formation. PMID- 16246093 TI - A role for actin in aging and apoptosis. AB - The actin cytoskeleton is central to many cell processes including membrane trafficking and generation of cell polarity. We have identified a role for actin in cell death and in promoting longevity of the budding yeast, Saccharomyces cerevisiae. Aging in yeast appears to occur via an apoptotic-like pathway with changes including DNA fragmentation, loss of mitochondrial membrane permeability, increase in levels of ROS (reactive oxygen species) and exposure of phosphatidylserine in the outer leaflet of the plasma membrane. This pathway can be induced by alterations in actin dynamics, such that reduced dynamics correlates with increased levels of ROS and decreased viability. Conversely, increased actin dynamics correlates with low ROS levels and increased survival. Our current studies have focused on identifying pathways which couple changes in actin dynamics to cell death. PMID- 16246094 TI - Arf, Sec7 and Brefeldin A: a model towards the therapeutic inhibition of guanine nucleotide-exchange factors. AB - GEFs (guanine nucleotide-exchange factors), which stimulate GDP dissociation from small G-proteins, are pivotal regulators of signalling pathways activated by small G-proteins. In the case of Arf proteins, which are major regulators of membrane traffic in the cell and have recently been found to be involved in an increasing number of human diseases, GDP/GTP exchange is stimulated by GEFs that carry a catalytic Sec7 domain. Recent structural results captured snapshots of the exchange reaction, revealing that Sec7 domains secure Arf-GDP to membranes before nucleotide exchange takes place, taking advantage of a built-in structural device in Arf proteins that couples their affinity for membranes to the nature of the bound nucleotide. One of the Arf-Sec7 intermediates was trapped by BFA (Brefeldin A), an uncompetitive inhibitor of Arf activation that has been instrumental in deciphering the molecular principles of membrane traffic at the Golgi. BFA targets a low-affinity Arf-Sec7 intermediate of the exchange reaction. It binds at the Arf-GDP/Sec7 interface, thus freezing the complex in an abortive conformation that cannot proceed to nucleotide dissociation. In the cell, this results in the specific inhibition of Arf1 by a subset of its GEFs, and the efficient and reversible block of membrane traffic at the Golgi. The mechanism of BFA leads to the concept of 'interfacial inhibition', in which a protein-protein interaction of therapeutic interest is stabilized, rather than impaired, by a drug. Up-regulated activity of small G-proteins is involved in various human diseases, making their GEFs attractive candidates to interrupt specifically the corresponding signalling pathway. Interfacial inhibitors are proposed as an alternative to competitive inhibitors that may be explored for their inhibition. PMID- 16246095 TI - Arf family GTPases: roles in membrane traffic and microtubule dynamics. AB - Database mining and phylogenetic analysis of the Arf (ADP-ribosylation factor) superfamily revealed the presence in mammals of at least 22 members, including the six Arfs, two Sars and 14 Arl (Arf-like) proteins. At least six Arf family members were found in very early eukaryotes, including orthologues of Arf, Sar, Arl2, Arl3, Arl6 and Arl8. While roles for Arfs in membrane traffic are well known, those for most of the Arls remain unknown. Depletion in cells of the most closely related human Arf proteins, Arf1-Arf5, reveals specificities among their cellular roles and suggests that they may function in pairs at different steps in endocytic and secretory membrane traffic. In addition, recent results from a number of laboratories suggest that several of the Arl proteins may be involved in different aspects of microtubule-dependent functions. Thus, a second major role for Arf family GTPases, that of regulating microtubules, is emerging. Because membrane traffic is often dependent upon movement of vesicles along microtubules this raises the possibility that these two fundamental functions of Arf family members, regulation of vesicle traffic and microtubule dynamics, diverged from one function of Arfs in the earliest cells that has continued to branch and allow additional levels of regulation. PMID- 16246096 TI - Interactions between Sla1p, Lsb5p and Arf3p in yeast endocytosis. AB - Endocytosis is critical for controlling the protein-lipid composition of the plasma membrane, uptake of nutrients as well as pathogens, and also plays an important role in regulation of cell signalling. While a number of pathways for endocytosis have been characterized in different organisms, all of these require remodelling of the cell cortex. The importance of a dynamic actin cytoskeleton for facilitating endocytosis has been recognized for many years in budding yeast, and is increasingly supported by studies in mammalian cells. Our studies have focused on proteins that we have shown to act at the interface between the actin cytoskeleton and the endocytic machinery. In particular, we have studied interactions of Sla1p, which binds to both activators of actin dynamics, i.e. Abp1p, Las17p and Pan1p, and to cargo proteins such as the pheromone receptor Ste2p. More recently we have mapped the interaction of Sla1p with Lsb5p, a protein that has a similar structure to the GGA [Golgi-localizing, gamma-adaptin ear homology domain, Arf (ADP-ribosylation factor)-binding] family of proteins with an N-terminal VHS (Vps27p/Hrs/STAM)-domain and a GAT (GGAs and TOM1) domain. We show that Lsb5p can interact with yeast Arf3p (orthologous with mammalian Arf6) and we demonstrate a requirement for Arf3p expression in order to localize Lsb5p to the cell cortex. PMID- 16246097 TI - Arfs, phosphoinositides and membrane traffic. AB - Arf (ADP-ribosylation factor) GTP-binding proteins function in cells to regulate membrane traffic and structure. Arfs accomplish this task through modification of membrane lipids and the recruitment of proteins, including coat proteins and actin, to membrane surfaces. Arf1 and Arf6 are the most divergent and most studied human Arf proteins that localize predominantly to the Golgi complex and plasma membrane respectively. We have been studying the targeting of Arf1 and Arf6 to these specific compartments and the common and divergent activities that they exert on these membranes. We have found that Arf6 acts through activation of type I phosphatidylinositol 4-phosphate 5-kinases to generate phosphatidylinositol 4,5-bisphosphate and that this activity is instrumental in facilitating the actin cytoskeletal rearrangements and alterations in endosomal membrane trafficking observed with increased Arf6 activation. Arf1 can also stimulate the activity of phosphatidylinositol kinases and recruit coat proteins and actin cytoskeletal elements to the Golgi complex. PMID- 16246098 TI - Centaurin-alpha1 and KIF13B kinesin motor protein interaction in ARF6 signalling. AB - The ARF (ADP-ribosylation factor) family of small GTPases regulate intracellular membrane trafficking by cycling between an inactive GDP- and an active GTP-bound form. Among the six known mammalian ARFs (ARF1-ARF6), ARF6 is the least conserved and plays critical roles in membrane trafficking and cytoskeletal dynamics near the cell surface. Since ARFs have undetectable levels of intrinsic GTP binding and hydrolysis, they are totally dependent on extrinsic GEFs (guanine nucleotide exchange factors) for GTP binding and GAPs (GTPase-activating proteins) for GTP hydrolysis. We have recently isolated a novel KIF (kinesin) motor protein (KIF13B) that binds to centaurin-alpha1, an ARF6GAP that binds to the second messenger PIP3 [PtdIns(3,4,5)P3]. KIFs transport intracellular vesicles and recognize their cargo by binding to proteins (receptors) localized on the surface of the cargo vesicles. Identification of centaurin-alpha1 as a KIF13B interactor suggests that KIF13B may transport ARF6 and/or PIP3 using centaurin-alpha1 as its receptor. This paper reviews the studies carried out to assess the interaction and regulation of centaurin-alpha1 by KIF13B. PMID- 16246099 TI - Centaurin beta4 in cancer. AB - Centaurin beta4 proteins are products of the DDEF1 (development and differentiation-enhancing factor 1) locus on human chromosome 8q24.1-24.2. Recent reports have indicated that this region and its products are amplified during development of several human cancers. Centaurins are GAPs (GTPase-activating proteins) that, together with GEFs (guanine nucleotide-exchange factors), regulate cyclic activation of Arfs (ADP-ribosylation factors), members of the Ras GTPase superfamily. Centaurin beta4 proteins associate with a variety of cellular signalling components implicated in control of growth, survival and movement and may act to direct assembly and/or disassembly of molecular complexes in concert with Arf, lipid and protein phosphorylation signalling pathways. PMID- 16246100 TI - Phosphoinositide regulation of clathrin-mediated endocytosis. AB - Endocytosis of transmembrane receptors largely occurs via clathrin-coated vesicles that bud from the plasma membrane and deliver their cargo to the endosomal system for recycling or degradation. PIs (phosphoinositides) control the timing and localization of endocytic membrane trafficking by recruiting adaptors and other components of the transport machinery, thereby being part of a coincidence detection system in adaptor-mediated vesicle transport. Activation of organelle- and substrate-specific PI kinases by small GTPases such as Arf (ADP ribosylation factor) and other factors may result in local changes of PI content, thereby regulating activity-dependent endocytic events including the recycling of synaptic vesicle membranes at nerve terminals. One such example is the PtdIns(4)P 5-kinase-mediated formation of PI(4,5)P2 [PtdIns(4,5)P2], which is required for the exo- and endo-cytic cycling of presynaptic vesicles and secretory granules. Over the last few years, protein X-ray crystallography in combination with biochemical and cell biological assays has been used to investigate the structure and function of many PI-binding proteins, including protein components of the endocytic machinery. These studies have provided molecular insights into the mechanisms by which PI(4,5)P2 recruits and activates adaptor proteins and their binding partners. In this mini-review, I will discuss the pathways of PI(4,5)P2 formation and its interactions with endocytic trafficking adaptors. PMID- 16246101 TI - Membrane traffic in cytokinesis. AB - A crucial facet of mammalian cell division is the separation of two daughter cells by a process known as cytokinesis. An early event in cytokinesis is the formation of an actomyosis contractile ring, which functions like a purse string in the constriction of the forming furrow between the cells. Far less well characterized are the membrane-trafficking steps which deliver new membrane to the cell surface during the plasma membrane expansion known to accompany furrow formation. It is now clearly established that the plasma membrane at the cleavage furrow of mammalian cells has a distinct lipid and protein composition from the rest of the plasma membrane. This may reflect a requirement for both increased surface area during furrowing and for the co-ordinated delivery of intracellular signalling or membrane re-modelling activities to the correct spatial coordinates during cleavage. In this review, we discuss recent work within the area of membrane traffic and cytokinesis. PMID- 16246102 TI - Retrograde signalling at the synapse: a role for Wnt proteins. AB - The formation of functional synapses requires a proper dialogue between incoming axons and their future synaptic targets. As axons approach their target, they are instructed to slow down and remodel to form proper presynaptic terminals. Although significant progress has been made in the identification of the mechanisms that control axon guidance, little is known about the mechanisms that regulate the conversion of actively growing axon into a presynaptic terminal. We found that Wnt secreted proteins are retrograde signals that regulate the terminal arborization of axons and synaptic differentiation. Wnts released from postsynaptic neurons induce extensive remodelling on incoming axons. This remodelling is manifested by a decrease in axon extension with a concomitant increase in growth-cone size. This morphological change is correlated with changes in the dynamics and organization of microtubules. Studies of a vertebrate synapse and the Drosophila neuromuscular junction suggest that a conserved Wnt signalling pathway modulates presynaptic microtubules as axons remodel during synapse formation. In this paper I discuss the role of the Wnt-Dvl (Dishevelled protein)-GSK-3beta (glycogen synthase kinase-3beta) signalling pathway in axon remodelling during synapse formation in the central nervous system. PMID- 16246103 TI - Physiological roles of spine motility: development, plasticity and disorders. AB - The vast majority of excitatory connections in the hippocampus are made on dendritic spines. Both dendritic spines and molecules within the membrane are able to move, but the physiological role of these movements is unclear. In the developing brain, spines show highly dynamic behaviour thought to facilitate new synaptic connections. Dynamic movements also occur in adults but the role of this movement is unclear. We have studied the effects of the most important excitatory neurotransmitter, glutamate, and found receptor activation to enhance movement of molecules within the spine membrane. This action of glutamate may be important in regulating the trafficking of neurotransmitter receptors that mediate change in synaptic function. In addition, we have studied the dynamic interactions between pre- and postsynaptic structures labelled with FM 4-64 and a membrane-targeted GFP (green fluorescent protein), respectively, in hippocampal slice cultures under conditions of increased activity, such as epilepsy. Our findings suggest a novel form of activity-dependent synaptic plasticity where spontaneous glutamate release is sufficient to trigger changes in the hippocampal microcircuitry by attracting neighbouring spines responsive to an enhanced level of extracellular glutamate. PMID- 16246104 TI - The regulation of membrane to cytosol partitioning of signalling proteins by phosphoinositides and their soluble headgroups. AB - Inositol phospholipids [PIs (phosphoinositides)] represent a group of membrane tethered signalling molecules which differ with respect to the number and distribution of monoester phosphate groups around the inositol ring. They function by binding to proteins which possess one of several domains that bind a particular PI species, often with high affinity and specificity. PH (pleckstrin homology) domains for example possess ligand-binding pockets that are often lined with positively charged residues and which bind PIs with varying degrees of specificity. Several PH domains bind not only PIs, but also their cognate headgroups, many of which occur naturally in cells as relatively abundant cytosolic inositol phosphates. The subcellular distributions of proteins possessing such PH domains are therefore determined by the relative levels of competing membrane-bound and soluble ligands. A classic example of the latter is the PH domain of phospholipase Cdelta1, which binds both phosphatidylinositol 4,5 bisphosphate and inositol 1,4,5-trisphosphate. We have shown that the N-terminal PH domain of the Rho family guanine nucleotide-exchange factor, Tiam 1, binds PI ligands promiscuously allowing multiple modes of regulation. We also recently analysed the ligand-binding specificity of the PH domain of PI-dependent kinase 1 and found that it could bind abundant inositol polyphosphates such as inositol hexakisphosphate. This could explain the dual distribution of this key signalling component, which needs to access substrates at both the plasma membrane and in the cytosol. PMID- 16246105 TI - Genetic, biochemical and structural approaches to talin function. AB - The cytoskeletal protein talin plays a key role in coupling the integrin family of cell adhesion molecules to the actin cytoskeleton. In this paper I present a brief review on talin and summarize our recent studies, in which we have taken both genetic and structural approaches to further elucidate the function of the protein. PMID- 16246106 TI - Two functionally distinct pools of Src kinases for PDGF receptor signalling. AB - The cytoplasmic tyrosine kinases of the Src family (SFK) play important roles in cell responses induced by growth factors, including cell growth, survival and migration. Here, we review how SFK participate in PDGF (platelet-derived growth factor) receptor signalling leading to DNA synthesis and actin assembly. Furthermore, evidence for a spatial compartmentalization of SFK signalling is also discussed. PMID- 16246107 TI - Compartmentalized cAMP signalling regulates vasopressin-mediated water reabsorption by controlling aquaporin-2. AB - The cAMP/PKA (protein kinase A) signalling pathway is activated by a plethora of stimuli. To facilitate the specificity of a cellular response, signal transduction complexes are formed and segregated to discrete sites (compartmentalization). cAMP/PKA signalling compartments are maintained by AKAPs (A-kinase anchoring proteins) which bind PKA and other signalling proteins, and by PDEs (phosphodiesterases). The latter hydrolyse cAMP and thus limit its diffusion and terminate PKA activity. An example of a cAMP-dependent process requiring compartmentalization of cAMP/PKA signals is arginine-vasopressin regulated water reabsorption in renal principal cells. A detailed understanding of the protein interactions within a signal transduction complex offers the possibility to design agents influencing PKA binding to a specific AKAP, the targeting of an AKAP or the interactions of AKAPs with other signalling molecules. The ability to specifically modulate selected branches of a signal transduction pathway would greatly advance basic research, and may lead to new drugs suitable for the treatment of diseases caused by dysregulation of anchored PKA signalling (e.g. renal and cardiovascular diseases). PMID- 16246108 TI - Compartmentalization of adenylate cyclase and cAMP signalling. AB - Concepts of cAMP signalling have changed dramatically from the linear cascades of just a few years ago, with the realization that numerous cellular processes affect this motif. These influences include other signalling pathways--most significantly Ca2+, scaffolding proteins (which are themselves variously regulated) to organize the elements of the pathway, and subcellular targeting of components. An obvious implication of this organization is that global measurements of cAMP may trivialize the complexity of the cAMP signals and obscure the regulation of targets. In this presentation, current developments on the targeting and assembly of ACs (adenylate cyclases) and their delivery to selected raft or non-raft domains of the plasma membrane will be discussed, along with the susceptibility of raft-targeted ACs to very discrete modes of increases in the intracellular Ca2+ concentration. Single-cell explorations of cAMP dynamics, as measured with cyclic nucleotide-gated channels, are also described in this paper, particularly as applied to cells in which the composition of AKAP (A-kinase anchoring protein)-PKA (protein kinase A)-PDE (phosphodiesterase) assemblies is probed by RNA interference ablation of defined AKAPs. PMID- 16246110 TI - Microtubule-associated proteins (MAPs) regulate cAMP signalling through exchange protein directly activated by cAMP (EPAC). AB - cAMP is an essential signalling molecule whose concentration in cells is regulated by a wide range of hormones. A large number of diseases, including cancer and asthma, are linked to improper regulation of the cAMP signalling system, and manipulation of cAMP levels by pharmaceutical agents has proven therapeutic benefit. The action of cAMP in cells is mediated through the signalling enzymes PKA (protein kinase A) and EPAC (exchange protein directly activated by cAMP). The study of the function of these proteins is essential to understand the role of cAMP in controlling disease. We have found that EPAC interacts with an ancillary protein, called LC2 (light chain 2), and this interaction enhances EPAC's ability to activate its substrate protein, Rap1 GTPase. This is an important finding because Rap1 is involved in the control of cell migration and cell shape, functions that are disrupted in diseases like cancer. LC2 appears to enhance EPAC activity towards Rap1 by increasing the ability of EPAC to interact with cAMP, so that EPAC activation occurs at lower concentrations of cAMP. The design of inhibitors that disrupt or enhance EPAC1 LC2 interaction may therefore form the basis of future therapeutics for diseases where cAMP signalling through Rap1 is improperly regulated. PMID- 16246109 TI - Imaging the cAMP-dependent signal transduction pathway. AB - In recent years, the development of new technologies based on the green fluorescent protein and FRET (fluorescence resonance energy transfer) has introduced a new perspective in the study of cAMP signalling. Real-time imaging of fluorescent biosensors is making it possible to visualize cAMP dynamics directly as they happen in intact, living cells, providing important and original insights for our understanding of the spatiotemporal organization of the cAMP/PKA (protein kinase A) signalling pathway. PMID- 16246111 TI - Cyclic nucleotide signalling: a molecular approach to drug discovery for Alzheimer's disease. AB - The EPAC (exchange protein directly activated by cAMP) proteins are GEFs (guanine nucleotide-exchange factors) that activate Rap GTPases upon binding to cAMP. The involvement of these proteins in a number of diseases, neurodegenerative, inflammatory and metabolic, has started to show how they may prove to be important targets for therapeutic intervention. We first became interested in EPAC when we discovered that the expression levels of both EPAC1 and EPAC2 were altered in those regions of the brain associated with Alzheimer's disease [McPhee, Breslin, Kewney, MacKenzie, Cooreman, Gibson and Hammond (2004) International Patent number WO 2004/096199 A2]. It was known that compounds could be designed to be selective for EPAC over PKA (protein kinase A); however, these compounds were all based around the core structure of cAMP. We decided to screen a small compound library (10000 compounds) to investigate the possibility of developing a compound series outside of the cAMP structure. We subsequently developed a novel, high-throughput screen based on the displacement of [3H]cAMP from the EPAC cAMP-binding site and identified small molecule hits from the Scottish Biomedical Lead Generation Library. These compounds selectively bind to the cAMP-binding sites of EPAC1 and EPAC2 and are structurally dissimilar to cAMP. They have similar affinities for both EPAC1 and EPAC2 and have a high degree of specificity for EPAC over PKA. We believe that these compounds provide a valuable starting point for a drug optimization programme. PMID- 16246112 TI - Beta-arrestin-recruited phosphodiesterase-4 desensitizes the AKAP79/PKA-mediated switching of beta2-adrenoceptor signalling to activation of ERK. AB - Using combined dominant-negative and siRNA (small interfering RNA)-mediated knockdown strategies, the functional importance of specific PDE4 (phosphodiesterase-4) isoforms in modifying signalling through the beta2-AR (beta2-adrenoceptor) has been uncovered. The PDE4D5 isoform preferentially interacts with the signalling scaffold protein beta-arrestin and is thereby recruited to the beta2-AR upon agonist challenge. Delivery of an active PDE to the site of cAMP synthesis at the plasma membrane specifically attenuates the activity of a pool of PKA (protein kinase A) that is tethered to the beta2-AR via AKAP79 (A-kinase anchoring protein 79). The specific functional role of this anchored PKA is to phosphorylate the beta2-AR and allow it to switch its coupling with G(i) and thereby activation of ERK (extracellular-signal-regulated kinase). Our studies uncover a novel facet of the regulation of beta2-AR signalling by showing that beta-arrestin-recruited PDE4 provides the means of desensitizing the agonist-dependent coupling of beta2-AR with G(i) and its consequential activation of ERK. PMID- 16246113 TI - Genetic manipulation and functional analysis of cAMP signalling in cardiac muscle: implications for a new target of pharmacotherapy. AB - Adenylate cyclase is a membrane-bound enzyme that catalyses the conversion of ATP into cAMP upon activation of cell-surface G-protein-coupled receptors, such as beta-adrenergic receptors, and initiates a cascade of phosphorylation reactions within the cell. Type 5 adenylate cyclase is a major isoform in the heart as well as in the striatum of the brain. Mice with a disrupted type 5 adenylate cyclase gene exhibited normal cardiac function under basal conditions, but a decreased response to isoprenaline stimulation. When mice were subjected to pressure overload stress with aortic banding, they developed cardiac hypertrophy, but with a significant reduction in the number of apoptotic cardiac myocytes as well as preserved cardiac function. When type 5 adenylate cyclase activity was inhibited pharmacologically, by the use of a novel P-site inhibitor with enhanced selectivity for this isoform, there were no changes in cardiac myocyte contractility, but the development of cardiac myocyte apoptosis induced by isoprenaline stimulation was effectively prevented. These results indicate that type 5 adenylate cyclase may serve as a better target of pharmacotherapy to prevent the development of cardiac myocyte apoptosis and thus failure in response to various cardiac stresses. PMID- 16246114 TI - Regulation of exocytosis by protein kinase C. AB - PKC (protein kinase C) has been known for many years to modulate regulated exocytosis in a wide variety of cell types. In neurons and neuroendocrine cells, PKC regulates several different stages of the exocytotic process, suggesting that these multiple actions of PKC are mediated by phosphorylation of distinct protein targets. In recent years, a variety of exocytotic proteins have been identified as PKC substrates, the best characterized of which are SNAP-25 (25 kDa synaptosome-associated protein) and Munc18. In the present study, we review recent evidence suggesting that site-specific phosphorylation of SNAP-25 and Munc18 by PKC regulates distinct stages of exocytosis. PMID- 16246115 TI - RIM function in short- and long-term synaptic plasticity. AB - RIM1alpha (Rab3-interacting molecule 1alpha) is a large multidomain protein that is localized to presynaptic active zones [Wang, Okamoto, Schmitz, Hofmann and Sudhof (1997) Nature (London) 388, 593-598] and is the founding member of the RIM protein family that also includes RIM2alpha, 2beta, 2gamma, 3gamma and 4gamma [Wang and Sudhof (2003) Genomics 81, 126-137]. In presynaptic nerve termini, RIM1alpha interacts with a series of presynaptic proteins, including the synaptic vesicle GTPase Rab3 and the active zone proteins Munc13, liprins and ELKS (a protein rich in glutamate, leucine, lysine and serine). Mouse KOs (knockouts) revealed that, in different types of synapses, RIM1alpha is essential for different forms of synaptic plasticity. In CA1-region Schaffer-collateral excitatory synapses and in GABAergic synapses (where GABA is gamma-aminobutyric acid), RIM1alpha is required for maintaining normal neurotransmitter release and short-term synaptic plasticity. In contrast, in excitatory CA3-region mossy fibre synapses and cerebellar parallel fibre synapses, RIM1alpha is necessary for presynaptic long-term, but not short-term, synaptic plasticity. In these synapses, the function of RIM1alpha in presynaptic long-term plasticity depends, at least in part, on phosphorylation of RIM1alpha at a single site, suggesting that RIM1alpha constitutes a 'phosphoswitch' that determines synaptic strength. However, in spite of the progress in understanding RIM1alpha function, the mechanisms by which RIM1alpha acts remain unknown. For example, how does phosphorylation regulate RIM1alpha, what is the relationship of the function of RIM1alpha in basic release to synaptic plasticity and what is the physiological significance of different forms of RIM-dependent plasticity? Moreover, the roles of other RIM isoforms are unclear. Addressing these important questions will contribute to our view of how neurotransmitter release is regulated at the presynaptic active zone. PMID- 16246117 TI - AMPA receptor phosphorylation during synaptic plasticity. AB - A widely studied example of vertebrate plasticity is LTP (long-term potentiation), the persistent synaptic enhancement that follows a brief period of coinciding pre- and post-synaptic activity. During LTP, different kinases, including CaMKII (calcium/calmodulin-dependent protein kinase II) and protein kinase A, become activated and play critical roles in induction and maintenance of enhanced transmission. Biochemical analyses have revealed several regulated phosphorylation sites in the AMPA (alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid) receptor subunits, GluR1 and GluR4. The regulated insertion of these receptors is a key event in the induction of LTP. Here, we discuss the phosphorylation of GluR1 and GluR4 and its role in receptor delivery and neuronal plasticity. PMID- 16246116 TI - Tyrosine phosphorylation of synaptophysin in synaptic vesicle recycling. AB - The integral SV (synaptic vesicle) protein synaptophysin was one of the first nerve terminal proteins identified. However its role, if any, in the SV life cycle remains undetermined. One of the most prominent features of synaptophysin is that its cytoplasmic C-terminus largely consists of pentapeptide repeats initiated by a tyrosine residue. Synaptophysin is heavily phosphorylated by tyrosine kinases in the nerve terminal, suggesting that this phosphorylation is central to its function. This review will cover the evidence for tyrosine phosphorylation of synaptophysin and how this phosphorylation may control its function in the SV life cycle. PMID- 16246118 TI - New metabolically stabilized analogues of lysophosphatidic acid: agonists, antagonists and enzyme inhibitors. AB - Lysophosphatidic acid (LPA) is a metabolically labile natural phospholipid with a bewildering array of physiological effects. We describe herein a variety of long lived receptor-specific agonists and antagonists for LPA receptors. Several LPA and PA (phosphatidic acid) analogues also inhibit LPP (lipid phosphate phosphatase). The sn-1 or sn-2 hydroxy groups have been replaced by fluorine, difluoromethyl, difluoroethyl, O-methyl or O-hydroxyethoxy groups to give non migrating LPA analogues that resist acyltransferases. Alkyl ether replacement of acyl esters produced lipase and acyltransferase-resistant analogues. Replacement of the bridging oxygen in the monophosphate by an alpha-monofluoromethylene-, alpha-bromomethylene- or alpha,alpha-difluoromethylenephosphonate gave phosphatase-resistant analogues. Phosphorothioate analogues with O-acyl and O alkyl chains are potent, long-lived agonists for LPA1 and LPA3 receptors. Most recently, we have (i) prepared stabilized O-alkyl analogues of lysobisphosphatidic acid, (ii) explored the structure-activity relationship of stabilized cyclic LPA analogues and (iii) synthesized neutral head group trifluoromethylsulphonamide analogues of LPA. Through collaborative studies, we have collected data for these stabilized analogues as selective LPA receptor (ant)agonists, LPP inhibitors, TREK (transmembrane calcium channel) K+ channel agonists, activators of the nuclear transcription factor PPAR-gamma (peroxisome proliferator-activated receptor-gamma), promoters of cell motility and survival, and radioprotectants for human B-cells. PMID- 16246119 TI - Critical role of acylglycerol kinase in epidermal growth factor-induced mitogenesis of prostate cancer cells. AB - The bioactive phospholipids, LPA (lysophosphatidic acid) and PA (phosphatidic acid), regulate pivotal processes related to the pathogenesis of cancer. Recently, we cloned a novel type of lipid kinase that phosphorylates monoacylglycerols (such as 2-arachidonoylglycerol, an endogenous cannabinoid receptor ligand) and diacylglycerols, to form LPA and PA, respectively. This AGK (acylglycerol kinase) is highly expressed in prostate cancer cell lines and the results reviewed here suggest that AGK might be a critical player in the initiation and progression of prostate cancer. Intriguingly, down-regulation of endogenous AGK inhibited EGF (epidermal growth factor), but not LPA-induced ERK1/2 (extracellular-signal-regulated kinase 1/2) activation and progression through the S-phase of the cell cycle. In this review, we will summarize the evidence demonstrating that AGK amplifies EGF growth signalling pathways that play an important role in the pathophysiology of prostate cancer. Because LPA has long been implicated as an autocrine and paracrine growth stimulatory factor for prostate cancer cells, the identification of this novel lipid kinase that regulates its production could provide new and useful targets for preventive or therapeutic measures. PMID- 16246120 TI - Structural characteristics of lysophosphatidic acid biological targets. AB - Lysophosphatidic acid (LPA; 1-acyl-3-phosphoglycerol) exerts its biological activity through both extracellular and intracellular targets. Receptor targets include the cell-surface G-protein-coupled receptors LPA(1-4) and the nuclear PPAR-gamma (peroxisome-proliferator-activated receptor gamma). Enzyme targets include the secreted cancer cell motility factor, autotaxin, and the transmembrane phosphatases, LPP1-3 (where LPP stands for lipid phosphate phosphatase). Ion channel targets include the two pore domain ion channels in the TREK family, TREK-1, TREK-2 and TRAAK. Structural features of these targets and their interactions with LPA are reviewed. PMID- 16246121 TI - Lipid phosphate phosphatases and lipid phosphate signalling. AB - Mammalian LPPs (lipid phosphate phosphatases) are integral membrane proteins that belong to a superfamily of lipid phosphatases/phosphotransferases. They have broad substrate specificity in vitro, dephosphorylating PA (phosphatidic acid), S1P (sphingosine 1-phosphate), LPA (lysophosphatidic acid) etc. Their physiological role may include the attenuation of S1P- and LPA-stimulated signalling by virtue of an ecto-activity (i.e. dephosphorylation of extracellular S1P and LPA), thereby limiting the activation of LPA- and S1P-specific G-protein coupled receptors at the cell surface. However, our recent work suggests that an intracellular action of LPP2 and LPP3 may account for the reduced agonist stimulated p42/p44 mitogen-activated protein kinase activation of HEK-293 (human embryonic kidney 293) cells. This may involve a reduction in the basal levels of PA and S1P respectively and the presence of an early apoptotic phenotype under conditions of stress (serum deprivation). Additionally, we describe a model whereby LPP2, but not LPP3, may be functionally linked to the phospholipase D1 derived PA-dependent recruitment of sphingosine kinase 1 to the perinuclear compartment. We also consider the potential regulatory mechanisms for LPPs, which may involve oligomerization. Lastly, we highlight many aspects of the LPP biology that remain to be fully defined. PMID- 16246122 TI - Thiol redox control via thioredoxin and glutaredoxin systems. AB - The Trx (thioredoxin) and Grx (glutaredoxin) systems control cellular redox potential, keeping a reducing thiol-rich intracellular state, which on generation of reactive oxygen species signals through thiol redox control mechanisms. Here, we give a brief overview of the human Trx and Grx systems. The main part focuses on our current knowledge about mitochondrial Grx2, which facilitates mitochondrial redox homoeostasis during oxidative stress-induced apoptosis. PMID- 16246123 TI - Oxidoreduction of protein thiols in redox regulation. AB - Protein cysteines can undergo various forms of oxidation, some of them reversible (disulphide formation, glutathionylation and S-nitrosylation). While in the past these were viewed as protein damage in the context of oxidative stress, there is growing interest in oxidoreduction of protein thiols/disulphides as a regulatory mechanism. This review discusses the evolution of the concept of redox regulation from that of oxidative stress and the redox state of protein cysteines in different cellular compartments. PMID- 16246124 TI - Defining the protein-protein interactions of the mammalian endoplasmic reticulum oxidoreductases (EROs). AB - The ER (endoplasmic reticulum) is the site of protein folding for all eukaryotic secreted and plasma membrane proteins. Disulphide bonds are formed in many of these proteins through a dithiol-disulphide exchange chain comprising two types of protein catalysts: PDI (protein disulphide-isomerase) and ERO (ER oxidoreductase) proteins. This review will examine what we know about ERO function, and will then consider ERO interactions and their implications for mammalian oxidative protein folding. PMID- 16246126 TI - Disulphide formation on mitochondrial protein thiols. AB - A large number of proteins contain free thiols that can be modified by the formation of internal disulphides or by mixed disulphides with low-molecular-mass thiols. The majority of these latter modifications result from the interaction of protein thiols with the endogenous glutathione pool. Protein glutathionylation and disulphide formation are of significance both for defence against oxidative damage and in redox signalling. As mitochondria are central to both oxidative damage and redox signalling within the cell, these modifications of mitochondrial proteins are of particular importance. In the present study, we review the mechanisms and physiological significance of these processes. PMID- 16246125 TI - Cell signalling by oxidized lipids and the role of reactive oxygen species in the endothelium. AB - The controlled formation of ROS (reactive oxygen species) and RNS (reactive nitrogen species) is now known to be critical in cellular redox signalling. As with the more familiar phosphorylation-dependent signal transduction pathways, control of protein function is mediated by the post-translational modification at specific amino acid residues, notably thiols. Two important classes of oxidant derived signalling molecules are the lipid oxidation products, including those with electrophilic reactive centres, and decomposition products such as lysoPC (lysophosphatidylcholine). The mechanisms can be direct in the case of electrophiles, as they can modify signalling proteins by post-translational modification of thiols. In the case of lysoPC, it appears that secondary generation of ROS/RNS, dependent on intracellular calcium fluxes, can cause the secondary induction of H2O2 in the cell. In either case, the intracellular source of ROS/RNS has not been defined. In this respect, the mitochondrion is particularly interesting since it is now becoming apparent that the formation of superoxide from the respiratory chain can play an important role in cell signalling, and oxidized lipids can stimulate ROS formation from an undefined source. In this short overview, we describe recent experiments that suggest that the cell signalling mediated by lipid oxidation products involves their interaction with mitochondria. The implications of these results for our understanding of adaptation and the response to stress in cardiovascular disease are discussed. PMID- 16246127 TI - What else has to happen for nitric oxide to induce cell death? AB - Nitric oxide (NO) can induce cell death; however, NO-induced cell death may be dependent/conditional on factors other than NO itself. Whether NO kills a particular cell depends on the amount of NO, source of NO, time of exposure to NO, cell type and the levels of other factors including, particularly oxygen, superoxide, H2O2, antioxidants, thiols and glycolysis. PMID- 16246128 TI - Role of GSH in the modulation of NOS-2 expression in the weaned mammary gland. AB - GSH delivery to the lactating mammary gland is essential for the maintenance of lactation as its decrease leads to apoptosis and involution of the mammary gland. In fact, it has already been demonstrated that some of the changes in gene expression found in the lactating mammary gland after forced weaning are reproduced in rats treated with buthionine sulphoximine to deplete GSH levels. An oligonucleotide microarray experiment would give us a better knowledge of the mRNA expression patterns during lactation and after weaning and the possible functions of GSH in the modulation of these events. PMID- 16246129 TI - Mitochondria and regulated tyrosine nitration. AB - The conditions of the cellular microenvironment in complex multicellular organisms fluctuate, enforcing permanent adaptation of cells at multiple regulatory levels. Covalent post-translational modifications of proteins provide the short-term response tools for cellular adjustment and growing evidence supports the possibility that protein tyrosine nitration is part of this cellular toolkit and not just a marker for oxidative damage. We have demonstrated that protein tyrosine nitration fulfils the major criteria for signalling and suggest that the normally highly regulated process may lead to disease upon excessive or inappropriate nitration. PMID- 16246131 TI - High-throughput subcellular protein localization using cell arrays. AB - Accomplishment of the human and mouse genome projects resulted in accumulation of extensive gene sequence information. However, the information about the biological functions of the identified genes remains a bottleneck of the post genomic era. Hence, assays providing simple functional information, such as localization of the protein within the cell, can be very helpful in the elucidation of its function. Transfected cell arrays offer a robust platform for protein localization studies. Open reading frames of unknown genes can be linked to a His6-tag or GFP (green fluorescent protein) reporter in expression vectors and subsequently transfected using the cell array. Cellular localization of the transfected proteins is detected either by specific anti-His-tag antibodies or directly by fluorescence of the GFP fusion protein and by counterstaining with organelle-specific dyes. The high throughput of the method in terms of information provided for every single experiment makes this approach superior to classical immunohistological methods for protein localization. PMID- 16246130 TI - Functional mapping of Toll/interleukin-1 signalling networks by expression cloning. AB - Multiple cellular proteins have been identified as participating in Toll/interleukin-1 receptor-mediated inflammatory gene expression. The continuing isolation of novel components, based on sequence similarities, protein-protein interactions and protein purification, suggests that many elements of this signalling network remain to be identified. We report here the development of a high-throughput functional screening platform and its application for the identification of components of inflammatory signalling networks. Our results enable us to estimate that 100-150 gene products are involved in controlling the transcription of the human interleukin 8 gene. The approach, which is simple and robust, constitutes a general method for mapping signal transduction systems and for rapid isolation of a large number of signalling components based on the control of pathways leading to regulation of gene expression. PMID- 16246132 TI - RNAi libraries and kinetoplast DNA. AB - African trypanosomes have a remarkable mitochondrial DNA termed kDNA (kinetoplast DNA) that contains several thousands of topologically interlocked DNA rings. Because of its highly unusual structure, kDNA has a complex replication mechanism. Our approach to understanding this mechanism is to identify the proteins involved and to characterize their function. So far approx. 30 candidate proteins have been discovered and we predict that there are over 100. To identify genes for more kDNA replication proteins, we are using an RNA interference library, which is the first forward genetic approach used for these parasites. PMID- 16246133 TI - Myocardial phenotyping using isotopomer analysis of metabolic fluxes. AB - Over the past 20 years, stable isotopes combined with isotopomer analysis have proven to be a powerful approach to probe the dynamics of metabolism in various biological systems, including the heart. The aim of this paper is to demonstrate how isotopomer analysis of metabolic fluxes can provide novel insights into the myocardial phenotype. Specifically, building on our past experience using NMR spectroscopy and GC-MS as applied to investigations of cardiac energy metabolism, we highlight specific complex metabolic networks that would not be predicted by classical biochemistry or by static measurements of metabolite, protein and mRNA levels. PMID- 16246134 TI - Ramifications of impaired PRPP synthesis in Saccharomyces cerevisiae. AB - The model eukaryote Saccharomyces cerevisiae is well suited to investigate the causes of metabolic disturbance. PRPP [5-phospho-D-ribosyl-1(alpha) pyrophosphate] may be regarded as a junction of carbon and nitrogen metabolism. As a result of this central position, perturbations in its synthesis can give rise to many unexpected cellular events, such as impaired cell integrity. We have taken advantage of S. cerevisiae's genetic tractability to investigate the metabolic links responsible for connecting the biochemical intermediate PRPP to apparently unrelated cellular functions. This approach provides insight into the co-ordination of different biological processes. PMID- 16246135 TI - Metabolic flux distribution analysis by 13C-tracer experiments using the Markov chain-Monte Carlo method. AB - Metabolic flux analysis using 13C-tracer experiments is an important tool in metabolic engineering since intracellular fluxes are non-measurable quantities in vivo. Current metabolic flux analysis approaches are fully based on stoichiometric constraints and carbon atom balances, where the over-determined system is iteratively solved by a parameter estimation approach. However, the unavoidable measurement noises involved in the fractional enrichment data obtained by 13C-enrichment experiment and the possible existence of unknown pathways prevent a simple parameter estimation method for intracellular flux quantification. The MCMC (Markov chain-Monte Carlo) method, which obtains intracellular flux distributions through delicately constructed Markov chains, is shown to be an effective approach for deep understanding of the intracellular metabolic network. Its application is illustrated through the simulation of an example metabolic network. PMID- 16246136 TI - Transcriptional regulation and metabolism. AB - Understanding organisms from a systems perspective is essential for predicting cellular behaviour as well as designing gene-metabolic circuits for novel functions. The structure, dynamics and interactions of cellular networks are all vital components of systems biology. To facilitate investigation of these aspects, we have developed an integrative technique called network component analysis, which utilizes mRNA expression and transcriptional network connectivity to determine network component dynamics, functions and interactions. This approach has been applied to elucidate transcription factor dynamics in Saccharomyces cerevisiae cell-cycle regulation, detect cross-talks in Escherichia coli two-component signalling pathways, and characterize E. coli carbon source transition. An ultimate test of system-wide understanding is the ability to design and construct novel gene-metabolic circuits. To this end, artificial feedback regulation, cell-cell communication and oscillatory circuits have been constructed, which demonstrate the design principles of gene-metabolic regulation in the cell. PMID- 16246137 TI - Modelling and simulation for metabolomics data analysis. AB - The advent of large data sets, such as those produced in metabolomics, presents a considerable challenge in terms of their interpretation. Several mathematical and statistical methods have been proposed to analyse these data, and new ones continue to appear. However, these methods often disagree in their analyses, and their results are hard to interpret. A major contributing factor for the difficulties in interpreting these data lies in the data analysis methods themselves, which have not been thoroughly studied under controlled conditions. We have been producing synthetic data sets by simulation of realistic biochemical network models with the purpose of comparing data analysis methods. Because we have full knowledge of the underlying 'biochemistry' of these models, we are better able to judge how well the analyses reflect true knowledge about the system. Another advantage is that the level of noise in these data is under our control and this allows for studying how the inferences are degraded by noise. Using such a framework, we have studied the extent to which correlation analysis of metabolomics data sets is capable of recovering features of the biochemical system. We were able to identify four major metabolic regulatory configurations that result in strong metabolite correlations. This example demonstrates the utility of biochemical simulation in the analysis of metabolomics data. PMID- 16246138 TI - Enhancing crop yield in Solanaceous species through the genetic manipulation of energy metabolism. AB - The improvement of crop yield has been endeavoured for centuries; whereas traditional breeding strategies have achieved this, until recently transgenic approaches to yield improvement have generally been less successful. In this mini review, we discuss metabolic engineering strategies specifically targeting energy metabolism as a strategy for yield enhancement. PMID- 16246139 TI - An in silico analysis of trypanosomatid RNA polymerases: insights into their unusual transcription. AB - African trypanosomes employ both Pol I (RNA polymerase I) and Pol II to transcribe protein-coding genes in large polycistronic units of up to 50 genes. Subsequent processing produces mature capped mRNAs. Evidence suggests that regulation of gene expression is primarily exerted post-transcriptionally. Here, we use the recently completed genome sequences of three trypanosomatids, Trypanosoma brucei, Trypanosoma cruzi and Leishmania major, in an in silico analysis of their fundamental RNA polymerase complexes. The core complement of Pol II subunits, including those that are shared with Pol I and Pol III are present. However, both Pol I and Pol III complexes are missing members of the rpoE-rpoF subunit groups. Out of the five shared subunits, both RPB5 and RPB6 have two isoforms in the three trypanosomes. One represents the canonical polymerase subunit and the other differs by insertion or deletion of stretches of charged residues. We propose that these alternative isoforms function in distinct polymerase complexes, and may influence recruitment of the trypanosome RPB4-RPB7 heterodimer. PMID- 16246140 TI - Chromosome positioning is largely unaffected in lymphoblastoid cell lines containing emerin or A-type lamin mutations. AB - Gene-poor human chromosomes are reproducibly found at the nuclear periphery in proliferating cells. There are a number of inner nuclear envelope proteins that may have roles in chromosome location and anchorage, e.g. emerin and A-type lamins. In the last decade, a number of diseases associated with tissue degeneration and premature aging have been linked with mutations in lamin A or emerin. These are termed laminopathies, with mutations in emerin causing Emery Dreifuss muscular dystrophy. Despite highly aberrant nuclear distributions of A type lamins and emerin in lymphoblastoid cell lines derived from patients with emerin or lamin A mutations, little or no change in chromosome location was detected. PMID- 16246141 TI - Nucleolar biogenesis: the first small steps. AB - The nucleolus is the site of rRNA transcription, pre-rRNA processing and ribosome subunit assembly. The nucleolus assembles around clusters of ribosomal gene repeats during late telophase, persists throughout interphase and then disassembles as cells enter mitosis. The initial step in nucleolar formation is ribosomal gene transcription, which is mediated by Pol I (RNA polymerase I) and its associated transcription factors: UBF (upstream-binding factor), SL1 (selectivity factor) and TIF-IA (transcription initiation factor IA)/Rrn3. Ribosomal gene clusters, termed NORs (nucleolar organizer regions), are found on each of the five human acrocentric chromosomes. Though transcription is repressed during metaphase, NORs that were active in the previous interphase form prominent cytogenetic features, namely secondary constrictions. The main defining characteristic of these constrictions is under-condensation in comparison with the rest of the chromosome. Extensive binding of UBF over the ribosomal gene repeat is responsible for the formation of this chromosomal feature. During interphase, the majority of the Pol I transcription machinery, though present in nucleoli, is not actively engaged in transcription. Interaction with UBF bound across the gene repeat provides an explanation for how this non-engaged Pol I machinery is sequestered by nucleoli. PMID- 16246142 TI - Transcriptional control by chromosome-associated protein phosphatase-1. AB - Tight regulation of gene expression is critical for cells to respond normally to physiological and environmental cues and to allow cell specialization. Reversible phosphorylation of key structural and regulatory proteins, from histones to the transcriptional machinery, is acknowledged to be an important mechanism of regulating spatial and temporal patterns of gene expression. PP1 (protein phosphatase-1), a major class of serine/threonine protein phosphatase, is found at many sites on Drosophila polytene chromosomes where it is involved in controlling gene expression and chromatin structure. PP1 is targeted to different chromosomal loci through interaction with a variety of different regulatory subunits, which modify PP1's activity towards specific substrates. This mini review gives an overview of known chromosome-associated PP1 complexes, their role in transcriptional control and the prospects for future analysis. PMID- 16246143 TI - Multifaceted role of the Saccharomyces cerevisiae Srs2 helicase in homologous recombination regulation. AB - Homologous recombination (HR) is a major pathway for the elimination of DNA DSBs (double-strand breaks) induced by high-energy radiation and chemicals, or that arise due to endogenous damage and stalled DNA replication forks. If not processed properly, DSBs can lead to cell death, chromosome aberrations and tumorigenesis. Even though HR is important for genome maintenance, it can also interfere with other DNA repair mechanisms and cause gross chromosome rearrangements. In addition, HR can generate DNA or nucleoprotein intermediates that elicit prolonged cell-cycle arrest and sometimes cell death. Genetic analyses in the yeast Saccharomyces cerevisiae have revealed a central role of the Srs2 helicase in preventing untimely HR events and in inhibiting the formation of potentially deleterious DNA structures or nucleoprotein complexes upon DNA replication stress. Paradoxically, efficient repair of DNA DSBs by HR is dependent on Srs2. In this paper, we review recent molecular studies aimed at deciphering the multifaceted role of Srs2 in HR and other cellular processes. These studies have provided critical insights into how HR is regulated in order to preserve genomic integrity and promote cell survival. PMID- 16246144 TI - Making crossovers during meiosis. AB - Homologous recombination (HR) is required to promote both correct chromosome segregation and genetic variation during meiosis. For this to be successful recombination intermediates must be resolved to generate reciprocal exchanges or 'crossovers' between the homologous chromosomes (homologues) during the first meiotic division. Crossover recombination promotes faithful chromosome segregation by establishing connections (chiasmata) between the homologues, which help guide their proper bipolar alignment on the meiotic spindle. Recent studies of meiotic recombination in both the budding and fission yeasts have established that there are at least two pathways for generating crossovers. One pathway involves the resolution of fully ligated four-way DNA junctions [HJs (Holliday junctions)] by an as yet unidentified endonuclease. The second pathway appears to involve the cleavage of the precursors of ligated HJs, namely displacement (D) loops and unligated/nicked HJs, by the Mus81-Eme1/Mms4 endonuclease. PMID- 16246145 TI - Roles of the Bloom's syndrome helicase in the maintenance of genome stability. AB - The RecQ family of DNA helicases is highly conserved in evolution from bacteria to humans. Of the five known human RecQ family members, three (BLM, WRN and RECQ4, which cause Bloom's syndrome, Werner's syndrome and Rothmund-Thomson syndrome respectively) are mutated in distinct clinical disorders associated with cancer predisposition and/or premature aging. BLM forms part of a multienzyme complex including topoisomerase IIIalpha, replication protein A and a newly identified factor called BLAP75. Together, these proteins play a role in the resolution of DNA structures that arise during the process of homologous recombination repair. In the absence of BLM, cells show genomic instability and a high incidence of sister-chromatid exchanges. In addition to a DNA structure specific helicase activity, BLM also catalyses Holliday-junction branch migration and the annealing of complementary single-stranded DNA molecules. PMID- 16246146 TI - Coupling ATP hydrolysis to DNA strand passage in type IIA DNA topoisomerases. AB - Type IIA topos (topoisomerases) catalyse topological conversions of DNA through the passage of one double strand through a transient break in another. In the case of the archetypal enzyme, DNA gyrase, it has always been apparent that the enzyme couples the free energy of ATP hydrolysis to the introduction of negative supercoiling, and the structural details of this process are now becoming clearer. The homologous type IIA enzymes such as topo IV and eukaryotic topo II also require ATP and it has more recently been shown that the energy of hydrolysis is coupled to a reduction of supercoiling or catenation (linking) beyond equilibrium. The mechanism behind this effect is less clear. We review the energy coupling process in both classes of enzyme and describe recent mechanistic and structural work on gyrase that addresses the mechanism of energy coupling. PMID- 16246148 TI - Helicases that interact with replication forks: new candidates from archaea. AB - Overcoming DNA replication fork blocks is essential for completing genome duplication and cell division. Archaea and eukaryotes drive replication using essentially the same protein machinery. Archaea may be a valuable resource for identifying new helicase components at advancing forks and/or in replication restart pathways. As described here, these may be relevant to understanding genome instability in metazoans. PMID- 16246147 TI - Recent advances in understanding structure-function relationships in the type II topoisomerase mechanism. AB - DNA topos (topoisomerases) are complex, multisubunit enzymes that remodel DNA topology. Members of the type II topo family function by passing one segment of duplex DNA through a transient break in another, a process that consumes two molecules of ATP and requires the co-ordinated action of multiple domains. Recent structural data on type II topo ATPase regions, which activate and enforce the directionality of DNA strand passage, have highlighted how ATP physically controls the catalytic cycle of the enzyme. Structural and biochemical studies of specialized DNA-binding domains in two paralogous bacterial type IIA topos (DNA gyrase and topo IV) show how these enzymes selectively negatively supercoil or decatenate DNA. Taken together, these findings expand our understanding of how disparate functional elements work together to co-ordinate the type II topo mechanism. PMID- 16246149 TI - Helicase-catalysed translocation and strand separation. AB - Helicases are molecular-motor enzymes that manipulate DNA or RNA during replication, repair, recombination, transcription, translation and processing of nucleic acids. The mechanisms for helicase activity have been studied intensely over the past decade. Recent advances in our understanding of the helicase mode of action have led to a general convergence of models that describe this diverse class of enzymes. One mechanism has been proposed that appears to have withstood the test of time, namely the inchworm mechanism. As the name implies, this mechanism involves a process whereby a helicase maintains at least two sites of contact with the nucleic acid. These binding sites can move relative to one another in a sequential fashion, resulting in net movement of the enzyme along the nucleic acid. The inchworm mechanism appears to be applicable to oligomeric states beyond the simple monomeric molecular motor. Although there are certainly many pertinent questions that remain unanswered, striking similarities in both form and function of seemingly disparate enzymes are becoming evident. PMID- 16246150 TI - Takeover of host ribosomes by divergent IRES elements. AB - The ribosome is the macromolecular machinery in the host cell for which all viruses have to compete. Early in infection, the viral mRNAs have to compete with the host for both the ribosomes and for the limited pool of eukaryotic initiation factors that are needed to facilitate the recruitment of ribosomes to both viral and cellular mRNAs. To circumvent this competition, certain viruses have evolved to recruit ribosomes to IRESs (internal ribosome entry sites), highly specialized RNA elements that are located at the 5'-end of the viral genomes. Here, we discuss how divergent IRES elements can recruit ribosomes and start protein synthesis with only a minimal set of eukaryotic translation initiation factors, and how this mode of translation initiation aids viral gene amplification during early onset of innate immune responses. PMID- 16246151 TI - Investigation of interactions of polypyrimidine tract-binding protein with artificial internal ribosome entry segments. AB - Most eukaryotic translation initiation is thought to be dependent on the 5'-cap structure of the mRNA. It is becoming apparent, however, that the mRNAs of many genes contain IRESs (internal ribosome entry segments) within the 5'-UTR (5' untranslated region) that allow ribosomes to initiate translation independently of the 5'-cap. IRESs can enable the expression of these genes under conditions (such as viral infection, cellular stress and apoptosis) when cap-dependent translation initiation is compromised, and also provide a target for regulation of gene expression. Recent results from our laboratory and others suggest that 10% of mRNAs (approximately 4000 genes) use this mechanism to initiate translation. One of the central goals of those working in the field of translation is to identify the sequence motif(s) and proteins that are required for internal ribosome entry. We have identified recently a unique PTB (polypyrimidine tract-binding protein) motif (CCU)n that is present in a large subset of cellular IRESs, and the results suggest that PTB itself is involved either directly or indirectly in ribosome recruitment. Here, we describe further investigations of PTB with artificial sequences that harbour this motif. PMID- 16246152 TI - eIF2B, a mediator of general and gene-specific translational control. AB - eIF2B (eukaryotic initiation factor 2B) is a multisubunit protein that is required for protein synthesis initiation and its regulation in all eukaryotic cells. Mutations in eIF2B have also recently been found to cause a fatal human disease called CACH (childhood ataxia with central nervous system hypomyelination) or VWM (vanishing white matter disease). This review provides a general background to translation initiation and mechanisms known to control eIF2B function, before describing molecular genetic and biochemical analysis of eIF2B structure and function, integrating work from studies of the yeast and mammalian eIF2B proteins. PMID- 16246153 TI - Unique features of selenocysteine incorporation function within the context of general eukaryotic translational processes. AB - Unlike other essential dietary trace elements, selenium exerts its biological actions through its direct incorporation into selenoproteins, as a part of the 21st amino acid, selenocysteine. Fundamental studies have elucidated the unique structures and putative functions of multiple co-translational factors required for the incorporation of selenocysteine into selenoproteins. The current challenge is to understand how these selenocysteine incorporation factors function within the framework of translation. In eukaryotes, co-ordinating nuclear transcription with cytoplasmic translation of genes is a challenge involving complex spatial and temporal regulation. Selenoproteins utilize the common cellular machinery required for synthesis of non-selenoproteins. This machinery includes the elements involved in transcription, mRNA splicing and transport, and translational processes. Many investigators have emphasized the differences between the expression of selenoproteins and other eukaryotic proteins, whereas this review will attempt to highlight common themes and point out where additional interactions may be discovered. PMID- 16246154 TI - Shoot development--genetic interactions in the meristem. AB - Development of the plant shoot is dependent on the shoot apical meristem. Interactions between KNOX homeodomain transcription factors and the myb domain transcription factor AS1 (ASYMMETRIC LEAVES1) regulate both meristem function as well as leaf patterning. This review summarizes these interactions. PMID- 16246155 TI - Amazing grass: developmental genetics of maize domestication. AB - Crop plants were domesticated by prehistoric farmers through artificial selection to provide a means of feeding the human population. This article discusses the developmental genetics of crop domestication and improvement, including the historical framework and recent approaches in maize and other grasses. In many cases, selecting for a plant form that correlates with productivity involves controlling meristem activity. In the domestication of modern maize from its progenitor Zea mays ssp. parviglumis, QTL (quantitative trait loci) mapping, genetics and population genomics approaches have identified several genes that contain signatures of selection. Only a few genes involved in the derivation of the highly productive maize ear have been identified, including teosinte glume architecture1 and ramosa1. Future prospects hinge on forward and reverse genetics, as well as on other approaches from the developing discipline of evo devo (evolutionary developmental biology). PMID- 16246156 TI - The regulation of cell migration by PTEN. AB - In vertebrates, the tumour suppressor PTEN (phosphatase and tensin homologue deleted on chromosome 10) regulates many cellular processes through its PtdIns(3,4,5)P3 lipid phosphatase activity, antagonizing PI3K (phosphoinositide 3 kinase) signalling. Given the important role of PI3Ks in the regulation of directed cell migration and the role of PTEN as an inhibitor of migration, it is somewhat surprising that data now indicate that PTEN is able to regulate cell migration independent of its lipid phosphatase activity. Here, we discuss the role of PTEN in the regulation of cell migration. PMID- 16246157 TI - The mechanism of hedgehog signal transduction. AB - Hedgehog (Hh) proteins are one of a small number of families of secreted signalling proteins that are responsible for cell interactions during development in many animals. As such, Hh signals produce many different responses at different times and in different cells. As for other multifunctional ligands, this requires regulated patterns of expression, special mechanisms for ligand movement between cells and ligand destruction, and mechanisms for integrating a generic signalling state (on or off) with the status of responding cells in order to produce an appropriate cell-specific response. Here I discuss what is known about the biochemical mechanisms by which an Hh signal is transduced in order to change the patterns of gene transcription. PMID- 16246158 TI - Myostatin: a modulator of skeletal-muscle stem cells. AB - Myostatin, or GDF-8 (growth and differentiation factor-8), was first identified through sequence identity with members of the BMP (bone morphogenetic protein)/TGF-beta (transforming growth factor-beta) superfamily. The skeletal muscle-specific expression pattern of myostatin suggested a role in muscle development. Mice with a targeted deletion of the myostatin gene exhibit a hypermuscular phenotype. In addition, inactivating mutations in the myostatin gene have been identified in 'double muscled' cattle breeds, such as the Belgian Blue and Piedmontese, as well as in a hypermuscular child. These findings define myostatin as a negative regulator of skeletal-muscle development. Myostatin binds with high affinity to the receptor serine threonine kinase ActRIIB (activin type IIB receptor), which initiates signalling through a smad2/3-dependent pathway. In an effort to validate myostatin as a therapeutic target in a post-embryonic setting, a neutralizing antibody was developed by screening for inhibition of myostatin binding to ActRIIB. Administration of this antimyostatin antibody to adult mice resulted in a significant increase in both muscle mass and functional strength. Importantly, similar results were obtained in a murine model of muscular dystrophy, the mdx mouse. Unlike the myostatin-deficient animals, which exhibit both muscle hypertrophy and hyperplasia, the antibody-treated mice demonstrate increased musculature through a hypertrophic mechanism. These results validate myostatin inhibition as a therapeutic approach to muscle wasting diseases such as muscular dystrophy, sarcopenic frailty of the elderly and amylotrophic lateral sclerosis. PMID- 16246159 TI - The homeodomain protein Nanog and pluripotency in mouse embryonic stem cells. AB - Intrinsic regulators of the pluripotency of mouse ES (embryonic stem) cells include the homeodomain proteins Oct4 and the recently identified Nanog. When overexpressed, Nanog displays the unique attribute of robustly sustaining ES cell self-renewal in the absence of the otherwise requisite extracellular stimulation by LIF (leukaemia inhibitory factor) and BMP (bone morphogenetic protein). Here, we review our current understanding of the function of Nanog in pluripotent stem cells both in vitro and in vivo. PMID- 16246160 TI - Role of the phosphoinositide 3-kinase pathway in mouse embryonic stem (ES) cells. AB - Mouse ES (embryonic stem) cells maintain pluripotency with robust proliferation in vitro. ES cells share some similarities with cancer cells, such as anchorage independent growth, loss of contact inhibition and tumour formation. After differentiation, ES cells lose pluripotency and tumorigenicity. Recent studies showed that the PI3K (phosphoinositide 3-kinase) pathway is important for proliferation, survival and maintenance of pluripotency in ES cells. The PI3K pathway is activated by growth factors and cytokines including insulin and leukaemia inhibitory factor. In addition to these exogenous factors, the PI3K pathway is endogenously activated by the constitutively active Ras family protein ERas (ES cell-expressed Ras). The PI3K pathway utilizes multiple downstream effectors including mTOR (mammalian target of rapamycin), which we have shown to be essential for proliferation in mouse ES cells and early embryos. PMID- 16246161 TI - Embryonic stem (ES) cells and embryonal carcinoma (EC) cells: opposite sides of the same coin. AB - Embryonal carcinoma (EC) cells are the stem cells of teratocarcinomas, and the malignant counterparts of embryonic stem (ES) cells derived from the inner cell mass of blastocyst-stage embryos, whether human or mouse. On prolonged culture in vitro, human ES cells acquire karyotypic changes that are also seen in human EC cells. They also 'adapt', proliferating faster and becoming easier to maintain with time in culture. Furthermore, when cells from such an 'adapted' culture were inoculated into a SCID (severe combined immunodeficient) mouse, we obtained a teratocarcinoma containing histologically recognizable stem cells, which grew out when the tumour was explanted into culture and exhibited properties of the starting ES cells. In these features, the 'adapted' ES cells resembled malignant EC cells. The results suggest that ES cells may develop in culture in ways that mimic changes occurring in EC cells during tumour progression. PMID- 16246162 TI - Cancer stem cells: AMLs show the way. AB - The blood-related cancer leukaemia was the first disease where human CSCs (cancer stem cells), or LSCs (leukaemic stem cells), were isolated. The haematopoietic system is one of the best tissues for investigating CSCs, since the developmental hierarchy of normal blood formation is well defined. Leukaemia can now be viewed as aberrant haematopoietic processes initiated by rare LSCs that have maintained or reacquired the capacity for indefinite proliferation through accumulated mutations and/or epigenetic changes. Yet, despite their critical importance, much remains to be learned about the developmental origin of LSCs and the mechanisms responsible for their emergence in the course of the disease. This report will review our current knowledge on LSC development and finally demonstrate how these discoveries provide a paradigm for identification of CSCs from solid tumours. PMID- 16246163 TI - Generation of embryos directly from embryonic stem cells by tetraploid embryo complementation reveals a role for GATA factors in organogenesis. AB - Gene targeting in ES (embryonic stem) cells has been used extensively to study the role of proteins during embryonic development. In the traditional procedure, this requires the generation of chimaeric mice by introducing ES cells into blastocysts and allowing them to develop to term. Once chimaeric mice are produced, they are bred into a recipient mouse strain to establish germline transmission of the allele of interest. Although this approach has been used very successfully, the breeding cycles involved are time consuming. In addition, genes that are essential for organogenesis often have roles in the formation of extra embryonic tissues that are essential for early stages of post-implantation development. For example, mice lacking the GATA transcription factors, GATA4 or GATA6, arrest during gastrulation due to an essential role for these factors in differentiation of extra-embryonic endoderm. This lethality has frustrated the study of these factors during the development of organs such as the liver and heart. Extraembryonic defects can, however, be circumvented by generating clonal mouse embryos directly from ES cells by tetraploid complementation. Here, we describe the usefulness and efficacy of this approach using GATA factors as an example. PMID- 16246164 TI - Role of MBD2 in gene regulation and tumorigenesis. AB - DNA methylation is an epigenetic mechanism involved in transcriptional silencing of imprinted genes, genes located on the inactive X chromosome, and a number of tumour suppressor genes in cancer. MBD (methyl-CpG-binding domain) proteins selectively bind to methylated DNA and recruit chromatin remodelling and transcriptional repressor complexes, thereby establishing a repressive chromatin state. MBD2, a member of the MBD protein family, binds to methylated promoter CpG islands (clusters of high-density CpG dinucleotides) and acts as a methylation dependent transcriptional repressor. Previous work has demonstrated that decreased CpG island methylation in mice lacking the DNA methyltransferase DNMT1 is associated with impaired tumorigenesis when crossed on the tumour-susceptible Apc(Min/+) background. Mbd2 deficiency also dramatically reduces adenoma burden and extends life span in a gene dosage-dependent manner in this mouse model. Mbd2 is therefore essential for tumorigenesis in the murine intestine, although it is dispensable for the viability of the host animals. These findings validate MBD2 as a potential target for therapeutic intervention in colorectal cancer. PMID- 16246165 TI - Post-transcriptional regulation in Xenopus embryos: role and targets of EDEN-BP. AB - EDEN (embryo deadenylation element)-dependent deadenylation is a regulatory process that was initially identified in Xenopus laevis early embryos and was subsequently shown to exist in Drosophila oocytes. Recent data showed that this regulatory process is required for somitic segmentation in Xenopus. Inactivation of EDEN-BP (EDEN-binding protein) causes severe segmentation defects, and the expression of segmentation markers in the Notch signalling pathway is disrupted. We showed that the mRNA encoding XSu(H) (Xenopus suppressor of hairless), a protein central to the Notch pathway, is regulated by EDEN-BP. Our data also indicate that other segmentation RNAs are targets for EDEN-BP. To identify new EDEN-BP targets, a microarray analysis has been undertaken. PMID- 16246166 TI - Contrasting mechanisms of regulating translation of specific Drosophila germline mRNAs at the level of 5'-cap structure binding. AB - Translational control is a key genetic regulatory mechanism underlying the initial establishment of the major spatial axes of the Drosophila embryo. Many translational control mechanisms target eIF4E (eukaryotic initiation factor 4E), an initiation factor that recognizes the 5'-cap structure of the mRNA. Cap recognition by eIF4E, in complex with eIF4G, is essential for recruitment of the mRNA to the small ribosomal subunit. One established mechanism for repressing translation involves eIF4E-binding proteins, which competitively inhibit the eIF4E-eIF4G interaction. Our group has uncovered a novel mechanism for repression in which an eIF4E cognate protein called d4EHP, which cannot bind eIF4G, binds to the 5'-cap structure of cad mRNA thus rendering it translationally inactive. These two related, but distinct, mechanisms are discussed and contrasted in this review. PMID- 16246167 TI - The life and death of translation elongation factor 2. AB - eEF2 (eukaryotic elongation factor 2) occupies an essential role in protein synthesis where it catalyses the translocation of the two tRNAs and the mRNA after peptidyl transfer on the 80 S ribosome. Recent crystal structures of eEF2 and the cryo-electron microscopy reconstruction of its 80 S complex now provide a substantial structural framework for dissecting the functional properties of this factor. The factor can be modified by either phosphorylation or ADP-ribosylation, which results in cessation of translation. We review the structural and functional properties of eEF2 with particular emphasis on the unique diphthamide residue, which is ADP-ribosylated by diphtheria toxin from Corynebacterium diphtheriae and exotoxin A from Pseudomonas aeruginosa. PMID- 16246168 TI - Coping with stress: eIF2 kinases and translational control. AB - In response to environmental stresses, a family of protein kinases phosphorylate eIF2 (eukaryotic initiation factor 2) to alleviate cellular injury or alternatively induce apoptosis. Phosphorylation of eIF2 reduces global translation, allowing cells to conserve resources and to initiate a reconfiguration of gene expression to effectively manage stress conditions. Accompanying this general protein synthesis control, eIF2 phosphorylation induces translation of specific mRNAs, such as that encoding the bZIP (basic leucine zipper) transcriptional regulator ATF4 (activating transcription factor 4). ATF4 also enhances the expression of additional transcription factors, ATF3 and CHOP (CCAAT/enhancer-binding protein homologous protein)/GADD153 (growth arrest and DNA-damage-inducible protein), that assist in the regulation of genes involved in metabolism, the redox status of the cells and apoptosis. Reduced translation by eIF2 phosphorylation can also lead to activation of stress-related transcription factors, such as NF-kappaB (nuclear factor kappaB), by lowering the steady-state levels of short-lived regulatory proteins such as IkappaB (inhibitor of NF kappaB). While many of the genes induced by eIF2 phosphorylation are shared between different environmental stresses, eIF2 kinases function in conjunction with other stress-response pathways, such as those regulated by mitogen-activated protein kinases, to elicit gene expression programmes that are tailored for the specific stress condition. Loss of eIF2 kinase pathways can have important health consequences. Mice devoid of the eIF2 kinase GCN2 [general control non derepressible-2 or EIF2AK4 (eIF2alpha kinase 4)] show sensitivity to nutritional deficiencies and aberrant eating behaviours, and deletion of PEK [pancreatic eIF2alpha kinase or PERK (RNA-dependent protein kinase-like endoplasmic reticulum kinase) or EIF2AK3] leads to neonatal insulin-dependent diabetes, epiphyseal dysplasia and hepatic and renal complications. PMID- 16246169 TI - TOPs and their regulation. AB - Upon cell-cycle arrest or nutrient deprivation, the cellular rate of ribosome production is reduced significantly. In mammalian cells, this effect is achieved in part through a co-ordinated inhibition of RP (ribosomal protein) synthesis. More specifically, translation initiation on RP mRNAs is inhibited. Translational regulation of RP synthesis is dependent on cis-elements within the 5'-UTRs (5' untranslated regions) of the RP mRNAs. In particular, a highly conserved 5'-TOP (5'-terminal oligopyrimidine tract) appears to play a key role in the regulation of RP mRNA translation. This article explores recent developments in our understanding of the mechanism of TOP mRNA regulation, focusing on upstream signalling pathways and trans-acting factors, and highlighting some interesting observations which have come to light following the recent development of cDNA microarray technology coupled with polysome analysis. PMID- 16246170 TI - IRES-dependent regulation of FGF-2 mRNA translation in pathophysiological conditions in the mouse. AB - The mRNA coding for FGF-2 (fibroblast growth factor 2), a major angiogenic factor, is translated by an IRES (internal ribosome entry site)-dependent mechanism. We have studied the role of the IRES in the regulation of FGF-2 expression in vivo, under pathophysiological conditions, by creating transgenic mice lines expressing bioluminescent bicistronic transgenes. Analysis of FGF-2 IRES activity indicates strong tissue specificity in adult brain and testis, suggesting a role of the IRES in the activation of FGF-2 expression in testis maturation and brain function. We have explored translational control of FGF-2 mRNA under diabetic hyperglycaemic conditions, as FGF-2 is implied in diabetes related vascular complications. FGF-2 IRES is specifically activated in the aorta wall in streptozotocin-induced diabetic mice, in correlation with increased expression of endogenous FGF-2. Thus, under hyperglycaemic conditions, where cap dependent translation is blocked, IRES activation participates in FGF-2 overexpression, which is one of the keys of diabetes-linked atherosclerosis aggravation. IRES activation under such pathophysiological conditions may involve ITAFs (IRES trans-acting factors), such as p53 or hnRNP AI (heterogeneous nuclear ribonucleoprotein AI), recently identified as inhibitory or activatory ITAFs respectively for FGF-2 IRES. PMID- 16246171 TI - The large spectrum of eIF2B-related diseases. AB - eIF2B (eukaryotic initiation factor 2B) is a GEF (guanine nucleotide-exchange factor) that plays, with its substrate eIF2, a key regulatory role in the translation initiation phase of protein synthesis. The importance of correct control of eIF2 and eIF2B for normal physiology is underlined by the recent involvement of the five genes that encode the five eIF2B subunits in a severe autosomal recessive neurodegenerative disease, described in young children as CACH (childhood ataxia with central nervous system hypomyelination)/VWM (leukoencephalopathy with vanishing white matter) syndrome. The syndrome is characterized by episodes of rapid deterioration during febrile illnesses or following head trauma and symmetrical demyelination of the brain white matter with cavitation aspects, leading to a progressive vanishing of the white matter replaced by CSF (cerebrospinal fluid). However, a wide clinical spectrum has been observed in the 148 patients presently reported, from congenital forms with rapid death to adult-onset forms with slow mental decline and progressive motor dysfunction, sometimes associated with congenital eye abnormalities or ovariodysgenesis. So far, 77 different mutations in each of the five EIF2B genes (EIF2B1-5), encoding subunits eIF2Balpha-epsilon, have been found, with two thirds affecting the eIF2Bepsilon subunit. The correlation found between the level of GEF activity of eIF2B in the mutated white blood cells and the age at disease onset suggests a direct role of the abnormal translation control in the pathophysiology of the disease. PMID- 16246172 TI - Control of mRNA stability in eukaryotes. AB - mRNA turnover plays a key role in the control of gene expression. Recent work has shown that proteins involved in mRNA turnover are located in multicomponent complexes which are tightly regulated. The control of mRNA stability is also intimately linked with translational processes. This article reviews the pathways and enzymes that control mRNA turnover in eukaryotic cells and discusses their mechanisms of control. PMID- 16246173 TI - Decapping the message: a beginning or an end. AB - Removal of the mRNA 5' cap is an important step in the regulation of mRNA stability. mRNAs are degraded by at least two distinct exonucleolytic decay pathways, one from the 5' end, and the second from the 3' end. Two major cellular decapping enzymes have been identified, and each primarily functions in one of the two decay pathways. The Dcp2 decapping enzyme utilizes capped mRNA as substrate and hydrolyses the cap to release m(7)GDP (N7-methyl GDP), while a scavenger decapping enzyme, DcpS, utilizes cap dinucleotides or capped oligonucleotides as substrate and releases m(7)GMP (N7-methyl GMP). In this review, we will highlight the function of different decapping enzymes and their role in mRNA turnover. PMID- 16246174 TI - Aberrant termination triggers nonsense-mediated mRNA decay. AB - NMD (nonsense-mediated mRNA decay) is a cellular quality-control mechanism in which an otherwise stable mRNA is destabilized by the presence of a premature termination codon. We have defined the set of endogenous NMD substrates, demonstrated that they are available for NMD at every round of translation, and showed that premature termination and normal termination are not equivalent biochemical events. Premature termination is aberrant, and its NMD-stimulating defects can be reversed by the presence of tethered poly(A)-binding protein (Pab1p) or tethered eRF3 (eukaryotic release factor 3) (Sup35p). Thus NMD appears to be triggered by a ribosome's failure to terminate adjacent to a properly configured 3'-UTR (untranslated region), an event that may promote binding of the UPF/NMD factors to stimulate mRNA decapping. PMID- 16246176 TI - Effect of lobe pumping on human albumin: investigating the underlying mechanisms of aggregate formation. AB - A common problem in the manufacture of liquid protein therapeutics is the tendency for aggregation and particle formation on extended storage. One aspect of processing that might contribute to particle formation is pumping. In the present study, we demonstrate that lobe pumps can promote aggregation in albumin preparations. This is accentuated where the clearance between the pump housing and lobes is increased. Under these conditions, the pump efficiency decreases, resulting in increased exposure of the protein to the pump environment. Depending on the inherent physicochemical stability of the protein, this can lead to aggregate formation, which can influence the long-term stability characteristics of the product. PMID- 16246177 TI - Effect of lobe pumping on human albumin: development of a lobe pump simulator using smoothed particle hydrodynamics. AB - Using SPH (smoothed particle hydrodynamics), the motion of a lobe pump under load was simulated in order to predict the level of shear stress experienced by a protein solution. By varying the gap size between the lobes and pump housing, variations in pump efficiency and shear stress were determined. The simulations indicated that pump shear was dependent on gap size, with shear stress levels (0 40 Pa) correlating with those estimated in previous albumin-pumping studies. As gap size increased, the number of fluid particles experiencing low shear (<10 Pa) increased, whereas those experiencing high shear (>20 Pa) showed a decreasing trend. The pump efficiency, however, decreased with gap size, requiring more lobe revolutions to pass a unit volume. Taken together, these observations indicate that lobe pumps operated with increased gaps between the rotors and the housing result in larger number of particles within the fluid experiencing shear stresses. Moreover, the simulations indicate that it is best to use larger lobe pumps operated at high efficiency to transfer protein-containing solutions. PMID- 16246175 TI - Biochemical characterization of RGS14: RGS14 activity towards G-protein alpha subunits is independent of its binding to Rap2A. AB - RGS (regulators of G-protein signalling) modulate signalling by acting as GAPs (GTPase-activating proteins) for alpha subunits of heterotrimeric G-proteins. RGS14 accelerates GTP hydrolysis by G(ialpha) family members through its RGS domain and suppresses guanine nucleotide dissociation from G(ialpha1) and G(ialpha3) subunits through its C-terminal GoLoco domain. Additionally, RGS14 binds the activated forms of the small GTPases Rap1 and Rap2 by virtue of tandem RBDs (Raf-like Ras/Rap binding domains). RGS14 was identified in a screen for Rap2 effectors [Traver, Splingard, Gaudriault and De Gunzburg (2004) Biochem. J. 379, 627-632]. In the present study, we tested whether Rap binding regulates RGS14's biochemical activities. We found that RGS14 activity towards heterotrimeric G-proteins, as either a GAP or a GDI (guanine nucleotide dissociation inhibitor), was unaffected by Rap binding. Extending our biochemical characterization of RGS14, we also examined whether RGS14 can suppress guanine nucleotide exchange on G(ialpha1) in the context of the heterotrimer. We found that a heterotrimer composed of N-myristoylated G(ialpha1) and prenylated G(betagamma) is resistant to the GDI activity of the GoLoco domain of RGS14. This is consistent with models of GoLoco domain action on free G(alpha) and suggests that RGS14 alone cannot induce subunit dissociation to promote receptor independent activation of G(betagamma)-mediated signalling pathways. PMID- 16246178 TI - Treatment of bleeding gastric varices. PMID- 16246179 TI - A review of juvenile polyposis syndrome. AB - Juvenile Polyposis Syndrome is an uncommon hamartomatous disorder with significant gastrointestinal malignant potential. Mutations in SMAD4 and BMPR1A, implicated in the Transforming Growth Factor beta pathway, have recently been characterized, and hold significance in the management of patients and at risk family members. This article reviews our knowledge to date of the genetics and clinicopathological features of the Juvenile Polyposis Syndrome, and discusses the current expert recommendations for genetic testing, disease screening and management. PMID- 16246180 TI - Hepatitis B virus infection in dialysis patients. AB - Hepatitis B virus (HBV) infection remains a major issue among dialysis patients. It is associated with a high risk of hepatic complication. The liver disease runs a unique clinical course in dialysis patients, as it can progress with modest hepatic inflammation and prominent fibrosis. The conventional cut-off level of serum alanine aminotransferase (ALT) for commencing antiviral therapy may prove too high and inappropriate for dialysis patients, and liver biopsy appears to be the only definitive means to establish the activity of liver disease in dialysis patients. Liver biopsy should be considered in patients with a serum ALT level that is persistently greater than 30 IU/L, or 0.75-fold the upper limit of the normal level, and/or other clinical and laboratory findings that suggest active liver disease. For antiviral treatment, preliminary reports have shown that lamivudine is effective and well tolerated in dialysis patients. However, the long-term efficacy of lamivudine and its optimal effective dose in dialysis patients remain unknown. The prevention of nosocomial transmission among dialysis patients is also important. Universal precaution measures should be strictly observed and the segregation of hepatitis B surface antigen-positive hemodialysis patients should be considered. For HBV non-immune patients, the importance of HBV vaccination should not be overemphasized. Until a new generation of highly immunogenic vaccines that are proven to be safe and effective in patients with end-stage renal disease becomes available, early vaccination before the development of end-stage renal failure remains the best way to secure immunological protection against HBV infection in dialysis patients. PMID- 16246181 TI - Improvement of gastrointestinal quality of life scores in cases of Helicobacter pylori-positive functional dyspepsia after successful eradication therapy. AB - Functional dyspepsia (FD) refers to a broad range of chronic upper abdominal symptoms associated with food intake. A definitive treatment for FD has not yet been established, and the effect of Helicobacter pylori (H. pylori) eradication still remains under debate. The Gastrointestinal Symptom Rating Scale (GSRS) is a specific questionnaire for patients with gastrointestinal symptoms. The present study examined the quality of life (QOL) of patients with H. pylori-positive FD following H. pylori eradication. METHODS: Sixty-eight patients with FD who gave informed consent were recruited for the study. H. pylori infection was diagnosed by the culture and histological methods, and the H. pylori eradication consisted of a 7-day course of lansoprazole, amoxicillin and clarithromycin. The overall success of the treatment was confirmed by a 13C urea breath test (UBT) conducted 3 months after the eradication. The GSRS questionnaire was administered to the patient just before the start of the eradication therapy and at 3 months after the therapy, just before the UBT was performed. RESULTS: In successfully eradicated patients, the total GSRS and the abdominal pain score significantly decreased. In particular, the abdominal pain score and indigestion score were significantly decreased after successful eradication in patients with ulcer-like FD or dysmotility-like FD. Conversely, in patients in whom the eradication was unsuccessful, neither the total GSRS nor any of the individual symptom scores showed any significant change. CONCLUSION: Successful H. pylori eradication improved the QOL of patients with FD, in particular H. pylori-positive patients with ulcer-like FD or dysmotility-like FD. PMID- 16246182 TI - Recovery of gastric function after Helicobacter pylori eradication in subjects with body atrophic gastritis: prospective 4-year study. AB - BACKGROUND: The relationship between Helicobacter pylori (H. pylori) eradication and atrophic changes in the gastric mucosa has not yet been fully defined. Although studies report a partial restoration of serum pepsinogen I (sPGI) levels after eradication, it is not clear if this finding reflects gastric mucosal healing on a morphological level. AIM: To assess alterations in gastric function after H. pylori eradication on moderate/severe body atrophic gastritis by determination of sPGI levels. METHODS: Twenty-three dyspeptic patients, selected from 284 consecutive H. pylori positive patients, with histological features of moderate/severe body atrophic gastritis and sPGI < 25 microg/L (11 men, mean age: 51.8 years, range: 29-79 years), underwent an upper gastrointestinal endoscopy with gastric biopsies and sPGI determination at baseline. All patients underwent eradication therapy. Serum pepsinogen I was measured again after 6 months, and at 1, 2, 3 and 4 years after eradication therapy. RESULTS: Mean sPGI levels prior to eradication were 11.9 microg/L (range: 4-23 microg/L). Six months after eradication therapy, mean sPGI levels significantly increased to 17.4 microg/L (P = 0.04). At the completion of the study, 4 years after eradication, sPGI levels increased from 17.4 to 32.7 microg/L (P = 0.01). A significant progressive increase in sPGI levels was observed from 6 months to 1 year (17.4 to 23.9 microg/L) and from 1 to 2 years (23.9 to 26.0 microg/L, P = 0.01). Serum pepsinogen I levels higher than the cut-off value of 25 microg/L were observed at various time-points: 6.3% of patients at 6 months (1/16), 33.3% (5/15) at 1 year, 50% (7/14) at 24 months, 66.7% (6/9) at 36 months and 87.5% (7/8) at 4 years. CONCLUSION: After H. pylori eradication, subjects with body atrophic gastritis showed long-term improvement of physiological gastric function, reflected by significantly and continually increasing sPGI levels over a 4-year period. PMID- 16246183 TI - Biochemical analysis of pancreatic fluid collections predicts bacterial infection. AB - BACKGROUND AND AIMS: Despite our understanding of the pathophysiology of different types of pancreatic fluid collections (PFC), few studies have attempted to correlate the biochemical analysis of PFC contents with clinical and radiological characteristics. The aim of this study was to assess the predictive value of fluid analysis for discerning collection type (pseudocyst vs acute fluid collection with necrosis), presence of infection or communication with the pancreatic duct in the setting of acute and chronic pancreatitis. METHODS: Pancreatic fluid from 34 consecutive patients undergoing endotherapy of PFC was prospectively analyzed for seven variables: lactate dehydrogenase (LDH), total protein, albumin, glucose, amylase, lipase and specific gravity. RESULTS: In multivariate analysis, adjusting for age and gender, high intracystic levels of protein (OR 6.2; 95% CI 1.3-37.0), LDH (OR 6.8 [2.3-38.3]), and albumin (OR 7.8 [1.3-67.4]), and low levels of glucose (OR 0.2 [0.03-0.9]) predicted the presence of PFC infection. The optimal threshold value for protein was 1000 g/dL, which achieved a sensitivity of 73% and specificity of 75% for detecting infection; the optimal cut-off for LDH was 1000 U/L (sensitivity 64%, specificity 85%), and the cut-off for albumin was 500 g/dL (sensitivity 75%, specificity 85%). There were no statistically significant differences in biochemical fluid analysis with respect to fluid collection type (pseudocysts vs acute fluid collection with necrosis) and the presence of pancreatic duct communication. CONCLUSIONS: Biochemical analysis of PFC fluid is clinically helpful in detecting fluid infection in patients with bacteria on Gram stain or positive fluid cultures. Our findings fail to support the utility of fluid analysis in characterizing cyst type, and we caution against its use in distinguishing pseudocysts from acute fluid collection with necrosis. PMID- 16246185 TI - Somatic mutations of mitochondrial DNA in digestive tract cancers. AB - BACKGROUND: Somatic mutations of mitochondrial DNA (mtDNA) have been reported to play an important role in the carcinogenesis of several human cancers. However, there are few reports on mtDNA mutations in digestive tract cancers, including esophageal, gastric and colorectal cancers. The present study examined somatic mtDNA mutations in these cancers. METHODS: Samples of 82 esophageal cancers, 96 gastric cancers and 138 colorectal cancers were collected. Mutations in the D310 mononucleotide repeat of mtDNA were examined by microsatellite assay. RESULTS: Frequencies of mtDNA mutations were similar in each digestive tract cancer: 14% (7/51) in esophageal cancers, 15% (14/94) in gastric cancers and 8% (11/133) in colorectal cancers. There were no significant relationships between mtDNA mutations and clinicopathological features, such as patient age or sex, tumor location, depth of tumor invasion and lymph node metastasis in each digestive tract cancer. CONCLUSIONS: The results suggest that mtDNA mutations play a role in the development but not progression in each digestive tract cancer, and that the role of mtDNA mutations might be similar among the digestive tract cancers. PMID- 16246184 TI - Bax and Bcl-2 protein expression in gastric precancerous lesions: immunohistochemical study. AB - BACKGROUND AND AIMS: Bcl-2 protein prolongs cell survival in the face of classical apoptotic stimuli, and is considered to be a suppressor of apoptosis. Bax plays a key role in apoptosis by accelerating cell death after an apoptotic stimulus. The aim of our study was to determine the roles of the Bax proapoptotic gene and the Bcl-2 antiapoptotic gene in the carcinogenesis of gastric cancer. METHODS: One hundred and forty-five gastric biopsy specimens of chronic gastritis, atrophic gastritis, intestinal metaplasia and gastric dysplasia were studied. Using immunohistochemical methods, Bax and Bcl-2 protein expression was observed. RESULTS: Bax was expressed in epithelial cells in all cases of chronic gastritis. Bax was not detected in 26% of specimens of atrophic gastritis. As intestinal metaplasia develops, Bax is further suppressed. In biopsy samples with dysplasia, Bax expression was demonstrated only in 12% of biopsy samples. Although Bcl-2 protein was not detected in chronic gastritis, aberrant expression was found in gastric epithelial intestinal metaplasia and dysplasia. CONCLUSIONS: The suppression of Bax and overexpression of Bcl-2 protein is an early event in gastric tumorigenesis, before gastric dysplastic changes occur. PMID- 16246186 TI - Pathogens causing community gastroenteritis in Australia. AB - BACKGROUND AND AIM: Many individuals with gastrointestinal symptoms do not seek medical attention and so there is little known about the pathogens involved in most cases of community gastroenteritis. We aimed to identify the pathogens responsible for community gastroenteritis and to examine the associated symptoms. METHODS: In a prospective study of 2811 subjects over 15 months, fecal pathogens were examined following highly credible gastroenteritis (HCG) events. The population consisted of family units of at least two children (< or =15 years old) and two adults each. Fecal samples were tested for a range of bacterial, viral and protozoal pathogens. Gastroenteric episode duration and symptoms such as vomiting, nausea and diarrhea were measured. RESULTS: One or more pathogens were identified in 198 of a total 791 specimens collected. Pathogens detected most often were Norovirus virus (10.7%), pathogenic E. coli (6.7%), Campylobacter spp. (3.0%) and Giardia sp. (2.5%). Children were more prone than adults to all the pathogens tested, except E. coli. Children infected with Campylobacter were 8.3 times more likely (95% CI: 2.7-25.4) to have a longer duration of diarrhea than children with Norovirus (P < 0.001). Similarly, children infected with E. coli had increased persistence of diarrhea compared to Norovirus (OR = 3.5; 95% CI: 1.3-9.5; P = 0.02). Infection with Norovirus in children meant greater persistence of vomiting symptoms than infection with Campylobacter (P = 0.005), E. coli (P = 0.03), or if no pathogen was identified (P = 0.004). Adults usually vomited for fewer days than children while duration of diarrhea was similar to children. CONCLUSIONS: Many of the pathogens responsible for cases of gastroenteritis in the Australian community are likely to go undetected by current surveillance systems and routine clinical practice. PMID- 16246187 TI - Inflammatory bowel disease in Iran: a review of 457 cases. AB - BACKGROUND AND AIMS: Inflammatory bowel disease (IBD) was believed to be infrequent in Iran; however, unofficial reports have confessed the continuing rise in IBD in our country. METHODS: Demographic and clinical features, extraintestinal manifestations, extension of disease and complications of 401 patients with ulcerative colitis (UC), 47 with Crohn's disease (CD), and nine with indeterminatn colitis (IC) were assessed retrospectively. The exact course of physicians' visits of 250 IBD patient was asked through face-to-face interview. RESULTS: Mean age at diagnosis was 31.9 years in UC and 30.5 years in CD patients. The male to female ratio was 0.8 for UC and 1.3 for CD. The percentage of CD and UC patients who were non-smokers was 82.9 and 84.5%, respectively. Patients with UC presented with rectal bleeding (41.9%), whereas those with CD complained of abdominal pain (46.9%). Among UC patients, proctosigmoid was affected in 51.9%. Colorectal cancer was diagnosed in two patients. The mean lag time between the onset of symptoms and definite diagnosis was 13.9 and 17.7 months for UC and CD patients, respectively. A total of 32.4% of patients with IBD had at least one of the five major extra-intestinal diseases. CONCLUSION: The demographic and clinical picture of IBD is more or less the same as that of other developing countries; however, the rarity of CD in Iran is noted. Although the true epidemiologic profile of IBD in Iran is still unknown, it is not as rare as previously thought, and it seems as if gradual adoption of a Western lifestyle may be associated with the continuing rise in IBD. PMID- 16246188 TI - Role of esophagogastroduodenoscopy in the initial assessment of children with inflammatory bowel disease. AB - INTRODUCTION: Diagnosis of inflammatory bowel disease (IBD) and differentiation between Crohn's disease (CD) and ulcerative colitis (UC) can be difficult in children. Several previous studies suggest that esophagogastroduodenoscopy (EGD) and biopsies are important in the initial investigation of children with suspected IBD. The aim of the present paper was to assess the importance of EGD in the initial diagnostic appraisal of children with suspected IBD. METHODS: Children diagnosed with IBD over a 4-year period were identified from a dedicated IBD database. Retrospective chart review documented presenting signs and symptoms, endoscopic features in the upper and lower gastrointestinal tract and histological findings on mucosal biopsies. RESULTS: Eighty-six children were diagnosed with IBD of whom 61 (70.9%) had CD, 13 (21.3%) UC, and the remainder, indeterminate colitis. Esophagogastroduodenoscopy was performed in 76 (88.4%). Nine children were diagnosed with IBD solely on the basis of information obtained following EGD. None of these children had colitis and all had abnormal histological findings on review of mucosal biopsies from the upper gastrointestinal tract. Thirteen (23.6%) of 55 children with CD had granulomas noted within biopsies obtained during EGD and another 20 had significant inflammatory changes on histological examination of upper gastrointestinal tract biopsies. Crohn's disease was diagnosed in 25 of 38 children with pan-colitis. Thirteen children were correctly classified as having CD only following assessment of their upper gastrointestinal tract. This included the presence of upper gut granulomata in eight children. CONCLUSION: The performance of EGD in these children with IBD provided additional diagnostic yield and guided the differentiation of disease type in many patients. Esophagogastroduodenoscopy is an essential component in the initial diagnostic assessment of children with possible CD or UC. PMID- 16246189 TI - Sclerotherapy for gastric fundal variceal bleeding: is complete obliteration possible without cyanoacrylate? AB - BACKGROUND: Many studies have suggested that endoscopic obliteration using cyanoacrylate for bleeding gastric fundal varices is effective. However, serious complications by injection of cyanoacrylate into varices have also been reported. METHODS: Thirty patients with bleeding gastric fundal varices underwent endoscopic injection sclerotherapy using 5% ethanolamine oleate under fluoroscopic guidance plus infusion of vasopressin and a transdermal nitroglycerin patch. The injection of 5% ethanolamine oleate was continued until it filled the varices and their feeder veins under fluoroscopic guidance. The injection needle was removed while thrombin glue was sprayed at the puncture site through the side hole of the injector needle to prevent bleeding from the puncture site. RESULTS: Complete hemostasis was achieved in 28/30 patients (93.3%). The cumulative rebleeding rate after 1, 3 and 5 years was 13%, 19% and 19%, respectively. The 1-, 3-, and 5-year cumulative mortality rates were 31%, 54% and 59%, respectively. There was no complication related to infusion of vasopressin and sclerotherapy procedure. CONCLUSION: The sclerotherapy method carried out using 5% ethanolamine oleate combined with infusion of vasopressin under fluoroscopic guidance might be a feasible method for obliteration of gastric fundal varices as an alternative to cyanoacrylate. PMID- 16246190 TI - Comparative study of diagnostic value of cytologic sampling by endoscopic ultrasonography-guided fine-needle aspiration and that by endoscopic retrograde pancreatography for the management of pancreatic mass without biliary stricture. AB - BACKGROUND: Endoscopic ultrasonography-guided fine-needle aspiration (EUS-FNA) can now provide a cytopathological diagnosis of underlying pancreatic malignancy with higher success rates than endoscopic retrograde pancreatography (ERP). To determine the significance of EUS-FNA for the diagnosis of pancreatic mass without biliary stricture, the value of cytopathological diagnosis obtained by EUS-FNA was retrospectively compared with that by ERP, and the complications associated with these procedures evaluated. METHODS: Eighty-three patients who were suspected to have a pancreatic mass (excluding a cystic mass), without biliary stricture on conventional ultrasound and/or computed tomography were enrolled. The EUS-FNA biopsy was performed in 53 patients and cytology utilizing ERP was performed in 30 patients. RESULTS: The sampling rate of adequate specimen was 100% in both groups. In the EUS-FNA group, the overall results for the available samples were sensitivity 92.9% and accuracy 94.3%. In contrast, in the ERCP group, the overall results were sensitivity 33.3% and accuracy 46.7%. There was a significant difference between the two groups (P < 0.01). With regard to complications, there was a significant difference (P < 0.01) in the frequency of post-procedure pancreatitis between the EUS-FNA group and ERP group (0%, 0/53 vs 33.3%, 10/30, respectively). CONCLUSION: Endoscopic ultrasonography-guided fine needle aspiration is safer and more accurate for the cytopathological diagnosis of suspected pancreatic masses without a biliary stricture as compared with cytology during ERP. Endoscopic ultrasonography with FNA should be considered a preferred test (prior to attempting endoscopic retrograde cholangiopancreatography) when a cytological diagnosis of a pancreatic mass is required, especially when there is no biliary obstruction, or when emergent decompression of an obstructed biliary tree is not considered clinically necessary due to lack of signs and symptoms of cholangitis. PMID- 16246191 TI - Community-based epidemiology of hepatitis B virus infection in West Bengal, India: prevalence of hepatitis B e antigen-negative infection and associated viral variants. AB - BACKGROUND AND AIMS: There is a paucity of population-based epidemiological information regarding hepatitis B virus (HBV) infection in India. The present study was planned to outline the magnitude and pattern of HBV infection, hepatitis B e antigen (HBeAg)-negative infection and the associated viral mutants in India. METHODS: A community-based epidemiological study of HBV infection was carried out in West Bengal, India. Serological markers of infection and potential risk factors for HBV transmission were determined. Among the infected individuals, HBV-DNA, genotypes and mutations in the precore (PC) stop codon and basal core promoter (BCP) regions were determined by DNA sequencing and polymerase chain reaction (PCR) restriction fragment length polymorphism methods. RESULTS: Of the 7653 people included in the study, 227 (2.97%) tested positive for hepatitis B surface antigen (HBsAg), of whom 204 (90%) were HBeAg-negative and hepatitis B e antibody (anti-HBe)-positive, and 78% had normal alanine aminotransferase (ALT) levels. HBV-DNA could be detected by PCR in only 32% of people. G1896A PC stop codon mutants were present in 12% of people, BCP mutants in 18% and the remainder (70%) of the HBeAg-negative infections were associated with wild type sequences in these regions. CONCLUSIONS: This first general population-based epidemiological study of HBV infection from India suggests that HBV acquisition starts in early childhood and peaks in adulthood. Most infections in the community are e-negative and inactive. The point prevalence of PC stop codon and BCP mutants is low in this primarily inactive and asymptomatic HBV infected population sample in eastern India. PMID- 16246192 TI - Interferon/long-term lamivudine combination therapy in anti-HBe positive chronic hepatitis B patients. AB - BACKGROUND: Monotherapy with a single antiviral agent is insufficient in controlling hepatitis B virus infection in the majority of patients with anti-HBe positive chronic hepatitis B. Interferon/long-term lamivudine combination therapy was evaluated to determine if this strategy would improve treatment efficacy and reduce the emergence of lamivudine resistance. METHODS: In total, 36 consecutive anti-HBe positive patients were treated with interferon (3 MU subcutaneously three times weekly) and lamivudine (100 mg orally once a day) for 12 months. After completion of the combined treatment, all patients continued to receive lamivudine monotherapy indefinitely. RESULTS: Overall, 35 patients (97%) showed virological response at 12 months. Four patients (11%) cleared HBsAg and developed anti-HBs. During the follow-up time, after the discontinuation of interferon, of 30 +/- 12 months (range: 7-57 months), 13 patients (36%) exhibited breakthrough infection. The cumulative rates of breakthrough infection at the end of 1, 2, 3 and 4 years of treatment were 0%, 14%, 32%, and 59%, respectively. CONCLUSIONS: Combination therapy appears to be effective and may also delay the selection of lamivudine-resistant variants. However, controlled trials are definitely warranted to clarify the potential benefits of combination antiviral treatment over monotherapy. PMID- 16246193 TI - Hepatitis B virus core and core-related antigen quantitation in Chinese patients with chronic genotype B and C hepatitis B virus infection. AB - BACKGROUND AND AIMS: Hepatitis B virus (HBV) core-related antigen (HBcrAg) and HBV core antigen (HBcAg) assays were developed for the measurement of serum HBV load. The aim of this study was to assess the clinical utility of these assays in Chinese patients with chronic genotype B and C HBV infection. METHODS: One hundred and ninety-three chronic hepatitis B patients were enrolled. Serum HBcrAg and HBcAg were measured by chemiluminescence enzyme immunoassay, and HBV-DNA was measured by using a sensitive polymerase chain reaction assay. The data were analyzed in patients with HBV genotype B (HBV/B) and genotype C (HBV/C). The HBcrAg/HBcAg ratio was calculated and compared between patients with and without hepatitis B e antigen (HBeAg). RESULTS: The concentrations of HBcrAg and HBcAg showed significant positive correlation with the HBV-DNA concentration in both HBV/B (r = 0.79, P < 0.001, and r = 0.77, P < 0.001, respectively) and HBV/C (r = 0.87, P < 0.001, and r = 0.90, P < 0.001, respectively). The cut-off for a positive HBcAg corresponded to approximately 4.5 log copies/mL, and that for a positive HBcrAg result corresponded to 3-4 log copies/mL. The HBcrAg/HBcAg ratio was higher in patients with HBeAg than in those without HBeAg. CONCLUSIONS: The HBcrAg assay and HBcAg assay are clinically useful in viral quantitation of HBV/B and HBV/C. A combination of these assays would be a valuable tool for analyzing the clinical status of HBV infection. PMID- 16246194 TI - Hepatitis C virus-NS3P in relation to p53, p21waf, mdm2, p21-ras and c-erbB2 in hepatocarcinogenesis. AB - BACKGROUND: The non-structural protein 3 (NS3P) of hepatitis C virus (HCV) genome was linked to the neoplastic transformation of normal hepatocytes in chronically infected patients. However, the exact mechanisms involved in this process are unidentified yet, especially in the Egyptian population where the commonest type is genotype 4. METHODS: We investigated 32 HCV reverse transcriptase-polymerase chain reaction (RT-PCR) positive hepatocellular carcinoma (HCC) cases and 18 morphologically normal hepatic tissues distant to tumors (MNT) for the correlation between HCV-NS3P, p53, p21(waf), mdm2, p21ras and c-erbB2 and DNA content by immunohistochemistry and image analysis. RESULTS: The NS3P expression was lower in HCC (65.6%) than in MNT (94.4%) patients. The expression level of studied genes in HCC was: p53 (56.25%), p21(waf) (43.7%), mdm2 (59.4%), p21-ras (73.3%) and c-erbB2 (75%). Whereas in MNT, it was 22.2, 61.1, 44.4, 41.2 and 77.8%, respectively. The NS3P expression showed a significant correlation with the presence of cirrhosis, chronic active hepatitis (CAH) and tumor grade (P < 0.05). c-erbB2 overexpression and p21(waf) loss were higher in MNT than in HCC patients, however, this did not reach a statistically significant level. There was a statistically significant correlation between NS3P, c-erbB2 and p21(waf) (P < 0.01). There was also a significant correlation between p21(waf) loss and CAH (P = 0.01) as well as between mdm2, c-erbB2 and cirrhosis (P = 0.025 and 0.001) in HCC cases. There was a statistically significant difference in the ploidy status between HCC and MNT, but there was no significant relationship between the ploidy status and other clinicopathological features. CONCLUSION: The carcinogenic effect of NS3P is probably exerted at an early stage of HCC possibly through a pathway involving c-erbB2 and p21(waf) alterations. In contrast, p53, p21ras and mdm2 alterations are late events in hepatocarcinogenesis and are usually associated with an aggressive phenotype. PMID- 16246195 TI - Psychological implications of hepatitis C virus diagnosis. AB - BACKGROUND AND AIMS: Hepatitis C virus (HCV) diagnosis causes significant psychological stress and anxiety. We thought it would be important to illustrate the anxiety caused by HCV diagnosis in patients from the developing world. METHODS: This study was conducted at the Shifa International Hospital, Islamabad, between February 2004 and April 2004. All patients who were recently diagnosed with HCV (those who tested positive to anti-HCV and HCV polymerase chain reaction) were given a questionnaire that compared stress due to HCV infection with four other variables, including death of a close family member, divorce, loss of source of income and move to another city. We also analyzed the anxiety level among these patients using the Beck Anxiety Inventory (BAI). RESULTS: We studied 98 patients and 100 healthy controls. Thirty-one (31.63%) patients who were diagnosed with HCV presented only with fatigue. Sixty-three (64.28%) patients admitted that diagnosis of HCV interfered with their daily life in some way. Among the newly diagnosed cases, 48 (48.97%) patients had moderate to severe anxiety (BAI Class B and C). Multi-logistic regression analysis showed that anxiety was related to HCV-related stress (P < 0.002) and self-perceived severity of disease (P < 0.001). HCV diagnosis was significantly more stressful than divorce (59.87 vs 70.95; P < 0.013), loss of source of income (50.52 vs 70.80; P < 0.001), and a move to another city (28.32 vs 70.80; P < 0.001). CONCLUSIONS: Diagnosis with HCV is reported to be more stressful than divorce, loss of source of income and a move to another city. PMID- 16246196 TI - Monitoring and management of antituberculosis drug induced hepatotoxicity. AB - BACKGROUND: Hepatotoxicity to antituberculosis therapy (ATT) poses a major challenge. This often results in inadequate therapy. The risk of fulminant hepatic failure and mortality is high once icteric hepatitis develops. There is no consensus on monitoring protocols and for the reintroduction of ATT. METHODS: All patients (from the Department of Internal Medicine and Gastroenterology, Jagjivanram Hospital and the Department of Gastroenterology, Bombay Hospital, Mumbai, India) with a diagnosis of tuberculosis, who were to receive ATT during the study period, were included in the present study for prospective periodic laboratory monitoring for the development of hepatotoxicity. Those patients who developed hepatotoxicity formed Group A (n = 21), whereas those who did not develop hepatotoxicity were included in Group C (n = 179). For the purpose of comparison with Group A, all the patients who presented directly with ATT induced hepatotoxicity during the study period were categorized as Group B (n = 24). Group A and B were further studied after normalization of liver functions for sequential reintroduction with therapeutic doses at a weekly interval. RESULTS: In Group A, 66.6% (14 patients) of the patients were diagnosed in the asymptomatic period. Seven patients had symptomatic hepatitis, but none had icteric illness. There were no mortalities in Group A. In contrast, all the patients in Group B had symptomatic hepatitis (75% icteric hepatitis). There was a mortality rate of 16.6% (four patients). Of the 41 patients from Groups A and B who survived, reintroduction was successful in 38/39 (97.4%). In the remaining two patients who were in Group B, reintroduction was not attempted because of decompensated liver disease. CONCLUSIONS: Periodic laboratory monitoring is important in detecting hepatotoxicity at an early stage, thereby preventing mortality. Sequential reintroduction is often successful. PMID- 16246197 TI - Hepatocyte growth factor accelerates the proliferation of hepatic oval cells and possibly promotes the differentiation in a 2-acetylaminofluorene/partial hepatectomy model in rats. AB - BACKGROUND: Hepatocyte growth factor (HGF) is the primary agent promoting the proliferation of mature hepatocytes. The purpose of the present paper was to clarify the effects of HGF on the proliferation and differentiation of hepatic oval cells using a 2-acetylaminofluorene/partial hepatectomy (2-AAF/PH) model in rats. METHODS: Recombinant human HGF (0.2 mg/day) was administered to 2-AAF/PH rats for 7 days using osmotic pumps intraperitoneally implanted in conjunction with hepatectomy (day zero). RESULTS: Periportal basophilic areas consisting of oval cells were significantly enlarged by treatment with HGF on day 8. In control animals, expression of alpha-fetoprotein (AFP) in the liver was gradually upregulated, leading a marked increase on day 12. In HGF-treated rats, AFP expression was stimulated at an earlier date and decreased to an undetectable level on day 12. Conversely, expression of albumin transcripts, which was stimulated by HGF-treatment at a later date, continued to increase even after HGF administration ceased, leading to an extremely high level on day 12. Moreover, treatment with HGF also stimulated the expression of hepatocyte nuclear factor 1alpha and -4alpha at an early date. CONCLUSIONS: These results indicate that, besides the proliferation of hepatic oval cells, HGF possibly promotes the differentiation to hepatocytes in vivo, suggesting that recombinant human HGF accelerates the regeneration of severely damaged livers, a situation in which the proliferation of mature hepatocytes is impaired. PMID- 16246198 TI - Lymphotoxin-beta production following bile duct ligation: possible role for Kupffer cells. AB - BACKGROUND AND AIMS: Lymphotoxin-beta (LT-beta) may play a role in the pathogenesis of chronic liver injury. The aim of this study was to determine in an animal model of bile duct ligation liver injury whether LT-beta expression is induced and whether Kupffer cells are an intrahepatic source of LT-beta. METHODS: Sprague-Dawley rats were divided into two groups: one group received a single dose of GdCl (a Kupffer cell-blocking agent, 10 mg/kg i.v.), whereas the other group received saline. One day later, the groups underwent bile duct ligation or a sham operation. Liver tissue was obtained on days 1, 3, 5, and 8 for assessment of Kupffer cell numbers, early fibrogenic events and LT-beta gene expression. Kupffer cells were isolated using pronase/collagenase perfusion and centrifugal elutriation. RESULTS: Hepatic LT-beta mRNA expression increased early following bile duct ligation. Pretreatment of bile duct-ligated animals with GdCl significantly reduced the number of Kupffer cells, delayed the rise in LT-beta expression, but had no effect on fibrogenesis. Recovery of the Kupffer cell population in these animals was accompanied by increased hepatic LT-beta expression. The LT-beta ligand and receptor were expressed by isolated normal Kupffer cells. CONCLUSIONS: Hepatic LT-beta expression is induced early following bile duct ligation. Kupffer cells may be an intrahepatic source of LT-beta. PMID- 16246200 TI - Clinical features of hepatocellular carcinoma with extrahepatic metastases. AB - BACKGROUND: There are few detailed clinical reports about extrahepatic metastases of hepatocellular carcinoma (HCC). The purpose of the present study was to elucidate the clinical features of extrahepatic metastases of HCC. METHODS: The clinical records of 482 patients who had been diagnosed as having HCC during the period from January 1995 to March 2001 were retrospectively reviewed. Extrahepatic metastases had been detected in 65 patients. Clinical features of those 65 patients were analyzed. RESULTS: Patients with extrahepatic metastases had more advanced intrahepatic tumors at the first diagnosis of HCC: 73.8% of the patients with extrahepatic metastases had tumors of intrahepatic tumor stage T3 or T4 according to the TNM classification, while only 28.5% of the patients without extrahepatic metastases had tumors of T3 or T4 (P < 0.001). Vessel invasion was also detected at the first diagnosis of HCC more frequently in the patients with extrahepatic metastasis (P < 0.001). The frequent metastatic sites were lung (53.8%), bone (38.5%), and lymph node (33.8%). Other metastatic sites were the adrenal gland, peritoneum, skin, brain and muscle. The median survival time and 1-year survival rate were 7 months (range: 1-59 months) and 24.9%, respectively. Patients with Child-Pugh grade B and C (P = 0.0018) and patients with positive serum alpha-fetoprotein (P = 0.011) had significantly poor prognosis. CONCLUSIONS: Extrahepatic metastases of HCC are not rare. The possibility of extrahepatic metastases and the clinical features of extrahepatic metastases should be considered when examining patients with HCC, particularly those with advanced intrahepatic tumors, to enable precise evaluation of the spread of HCC and determination of the appropriate treatment method. PMID- 16246201 TI - Images of interest. Gastrointestinal: Boerhaave's syndrome. PMID- 16246199 TI - Distribution and isoforms of epimorphin in carbon tetrachloride-induced acute liver injury in mice. AB - BACKGROUND AND AIM: Epimorphin, a morphoregulatory factor essential to organ development, is believed to direct normal morphogenesis in tissue repair. We examined the dynamics and the roles of epimorphin, a cell surface-associated molecule detected on mesenchymal cells, in hepatic tissue repair from acute liver injury. METHODS: After acute liver injury was induced by carbon tetrachloride in Balb/c mice, the distribution of epimorphin-expressing cells was studied immunohistochemically. To clarify interactions between epimorphin expression and hepatocyte behavior, epimorphin-expressing cells and proliferating hepatocytes were counted. Then, epimorphin quantity and isoforms were assessed by western blotting. To better understand effects of epimorphin, we cultured rat hepatocytes in its presence. RESULTS: Epimorphin was distributed in relation to sinusoids, portal veins, central veins and granulomas, expressed in stellate cells and myofibroblasts. In the periportal zone, the expression in sinusoids was decreased at 24 h but increased on day 7 after carbon tetrachloride administration. Numbers of epimorphin-expressing cells and proliferating hepatocytes changed in an inverse manner as time progressed. In the pericentral zone, reactivity for epimorphin was markedly enhanced concurrently with appearance of granulomas. Quantities of 34-kDa isoform paralleled epimorphin-staining intensity. In vitro, epimorphin induced spherical hepatocyte aggregates and maintained differentiated hepatocyte function. CONCLUSIONS: Epimorphin is involved in tissue repair following a single injection of carbon tetrachloride, in which distribution and the quantity of epimorphin expression are important, particularly in maintaining hepatocyte function. PMID- 16246202 TI - Images of interest. Gastrointestinal: Campylobacter enterocolitis. PMID- 16246203 TI - Images of interest. Gastrointestinal: Kirklin's sign. PMID- 16246204 TI - Images of interest. Gastrointestinal: small bowel inflammation and ulcerative colitis. PMID- 16246205 TI - Susceptibility genes and functional gastrointestinal disorders. PMID- 16246206 TI - Large regenerative nodule perfused by the portal vein. AB - In a 42-year-old Japanese woman with esophageal varices, abdominal ultrasound (US) demonstrated a hyperechoic lesion 3 cm in diameter in segment 4 (S4). This nodular lesion had high intensity on T1-weighted magnetic resonance imaging (MRI), low intensity on T2, and very high intensity on superparamagnetic iron oxide (SPIO) enhanced MRI. Angiography showed sparse distribution of arterial branches and dense distribution of portal branches in S4. Meandering, thin arteries were seen in the peripheral area of the right lobe. The second branches of the portal vein were hardly visualized anywhere in the liver. Computed tomography arterioportography (CTAP) revealed portal blood flow dominance in this nodular lesion. There was no evidence of ischemic liver damage, such as thromboembolic episodes, laboratory data of liver damage, coagulation abnormalities etc. Therefore this abnormality was more likely to be caused by anomalous changes than thrombotic changes. Needle biopsy revealed no atypical cells. Two years later, the nodule size was reduced to 1.9 cm, showing its benign nature. Based on these findings, this lesion was classified as a new type of large regenerative nodule (LRN) associated with anomalies in the portal veins and arteries. This is the first report of an LRN of this size in which portal vein perfusion was dominant. Moreover, this lesion was difficult to differentiate from hepatocellular carcinoma (HCC) by imaging. Analysis of the images and pathological features of this case would contribute to a better understanding of the pathogenesis of nodular lesions of the liver. PMID- 16246207 TI - Xanthoma striatum palmare in a patient with primary biliary cirrhosis and hypercholesterolemia. PMID- 16246208 TI - Primary ileal villous atrophy associated with microscopic colitis. PMID- 16246209 TI - Obstructive jaundice and pancreatitis secondary to percutaneous endoscopic gastrostomy tube migration. PMID- 16246210 TI - Long-term anticoagulation therapy for a cirrhotic patient with recurrent deep venous thrombosis. PMID- 16246212 TI - Prevalence of obesity in Taiwan. AB - In Taiwan, the prevalence and problem of obesity has increased significantly in recent decades and has became an important public health issue. In children (12 15 years), the prevalence of obesity (defined as body weight > 120% of mean body weight with age- and gender-specification) was 12.4%, 14.8% and 15.6% among boys and 10.1%, 11.1% and 12.9% among girls in 1980, 1986 and 1996 respectively. A survey of 1500 12-15-year-old children during 1995-1996 in Taipei city found that about 16.6% of boys and 11.1% of girls were obese, while an additional 11.6% of boys and 10.2% of girls were overweight. In adults, using the criteria defined by the Department of Health in Taiwan [overweight as (body mass index) BMI > or = 24 and obese as BMI > or = 27], the age-adjusted prevalence of obesity was 10.5% and 15.9% for men and 13.2% and 10.7% for women from 1993-1996 to 2000-2001. From these data, we found that the prevalence of obesity in Taiwan has increased steadily from 1980 to 2000 especially in children and in men. PMID- 16246213 TI - Obesity and the electrocardiogram. AB - Obesity is associated with a wide variety of electrocardiographic (ECG) abnormalities. Most of these reflect alterations in cardiac morphology. Some serve as markers of risk for sudden death. Key ECG abnormalities or alterations occurring with disproportionately high frequency in obese subjects include: leftward shifts of the P wave QRS and T wave axes, various changes in P wave morphology, low QRS voltage, various markers of left ventricular hypertrophy (particularly the Cornell voltage and product), T wave flattening in the inferior and lateral leads, lengthening of the corrected QT interval and prolonged QT interval duration. Alterations in the signal-averaged ECG and in heart rate variability may be arrhythmogenic. Cardiac arrhythmias have been described in obese subjects, but are often accompanied by left ventricular hypertrophy or the sleep apnea syndrome. Many of these ECG abnormalities are reversible with substantial weight loss. Thus, obesity is associated with a wide variety of ECG abnormalities, many of which are corrected by weight loss. PMID- 16246214 TI - Diagnosis and management of the metabolic syndrome in obesity. AB - The metabolic syndrome is a constellation of interrelated abnormalities that increase the risk for cardiovascular disease and progression to type 2 diabetes. The prevalence of this syndrome is increasing because of the 'obesity epidemic'. The National Cholesterol Education Program Adult Treatment Panel III defined practical criteria for the diagnosis of the metabolic syndrome and established the basic principles for its management. Also, the International Diabetes Federation recently proposed another definition. The metabolic syndrome is a secondary target for cardiovascular risk reduction. Clinicians should identify individuals with this condition, assess their cardiovascular risk and treat them by an aggressive and multifaceted approach. The most effective therapeutic intervention in patients with the metabolic syndrome should focus on modest weight reduction and regular physical activity. Adoption of a healthier diet and smoking cessation are necessary. Drug therapy may be needed to achieve recommended goals if therapeutic lifestyle changes are not sufficient. Low density lipoprotein cholesterol is the primary target of therapy (new aggressive goals should be achieved). Statins are probably the drugs of choice. Fibrates and nicotinic acid are also useful options. Hypertension should be managed aggressively probably starting with an inhibitor of the renin-angiotensin system or a calcium channel blocker and adding a low dose of a thiazide diuretic if necessary. Aspirin should be administered if the cardiovascular risk is high. In the future acarbose, metformin, meglitinides and thiazolidinediones may be used in patients with the metabolic syndrome to delay the onset of type 2 diabetes and reduce cardiovascular risk. Such an intense and multifactorial approach is likely to reverse the bad prognosis associated with the metabolic syndrome. PMID- 16246215 TI - Role of cholecystokinin in appetite control and body weight regulation. AB - Summary Cholecystokinin (CCK), a peptide that is distributed widely throughout the gastrointestinal tract and the central nervous system, has a number of physiological effects including the stimulation of gallbladder contraction and pancreatic and gastric acid secretion, slowing of gastric emptying and suppression of energy intake. This review focuses on current knowledge relating to (i) the effects of CCK on energy intake; (ii) the role for CCK in the pathophysiology of obesity; and (iii) the therapeutic potential for strategies which modulate the action or secretion of CCK in the management of obesity. While CCK plays a role in the acute regulation of appetite and energy intake, there is little evidence to suggest that specific CCK receptor agonists, or modulation of the actions of endogenous CCK by dietary manipulation, have sustainable inhibitory effects on energy intake. Hence, it appears unlikely that manipulating the pathways by which CCK modulates energy intake will prove to be an effective strategy in the long term management of obesity. PMID- 16246217 TI - Orlistat for the treatment of obesity: cost utility model. AB - This study aimed to assess the cost utility of orlistat treatment based on (i) criteria from recent guidance from the National Institute for Clinical Excellence (NICE) for England and Wales (treatment discontinued if weight loss < 5% at 3 months; and < 10% at 6 months); and (ii) alternative criteria from the European Agency for the Evaluation of Medicinal Products (EMEA) licence for orlistat prescription in the European Community (treatment discontinued if weight loss < 5% at 3 months). Subjects were 1398 obese individuals who participated in three large European Phase III trials of orlistat treatment for adults (BMI: 28-47 kg m(-2)). Measures were: response to treatment in orlistat and placebo treatment groups; health benefit expressed as quality adjusted life years (QALYs) gained associated with weight loss; costs associated with orlistat treatment. In the cost utility model with multiway sensitivity analysis, the cost/QALY gained using the NICE criteria was estimated to be 24,431 pounds (sensitivity analysis range: 10,856 to 77,197 pounds). The cost/QALY gained using the alternative EMEA criteria was estimated to be 19,005 pounds (range: 8,840 to 57,798 pounds). In conclusion, NICE guidance for the continued use of orlistat was supported in this updated cost utility model, comparing favourably with a previously published estimate of 45,881 pounds per QALY gained. Moreover, the value for money of orlistat treatment is improved further if EMEA treatment criteria for continued orlistat treatment are applied. The EMEA criteria should be considered in any future changes to the NICE guidance or in guidance issued by similar agencies. PMID- 16246216 TI - PYY3-36 as an anti-obesity drug target. AB - The neuropeptide Y (NPY)/peptide YY (PYY) system has been implicated in the physiology of obesity for several decades. More recently ignited enormous interest in PYY3-36, an endogenous Y2-receptor agonist, as a promising anti obesity compound. Despite this interest, there have been remarkably few subsequent reports reproducing or extending the initial findings, while at the same time studies finding no anti-obesity effects have surfaced. Out of 41 different rodent studies conducted (in 16 independent labs worldwide), 33 (83%) were unable to reproduce the reported effects and obtained no change or sometimes increased food intake, despite use of the same experimental conditions (i.e. adaptation protocols, routes of drug administration and doses, rodent strains, diets, drug vendors, light cycles, room temperatures). Among studies by authors in the original study, procedural caveats are reported under which positive effects may be obtained. Currently, data speak against a sustained decrease in food intake, body fat, or body weight gain following PYY3-36 administration and make the previously suggested role of the hypothalamic melanocortin system unlikely as is the existence of PYY deficiency in human obesity. We review the studies that are in the public domain which support or challenge PYY3-36 as a potential anti-obesity target. PMID- 16246218 TI - Hippocrates: 'To talk, move during conversation, think and listen to others increases the combustion'. PMID- 16246221 TI - Etiology and prevention of gastric cancer: a population study in a high risk area of China. AB - A series of studies has been carried out in Linqu County, Shandong Province, China, a high-risk area for gastric cancer, to investigate the risk factors associated with gastric cancer, precancerous lesions and the prevention of gastric cancer. Our studies showed that sour pancakes (a popular local food), salted foods, cigarette smoking, and family history of gastric cancer were risk factors, whereas fresh vegetables, and intake of vitamin C and calcium were inversely associated with the risk of gastric cancer. The prevalence of chronic atrophic gastritis was approximately 20% in an adult population in Linqu County, intestinal metaplasia was approximately 50%, and dysplasia was approximately 20%. A follow-up study showed that the relative risk of developing gastric cancer increased with the severity of gastric lesions, and was associated with dietary factors, cigarette smoking and H. pylori infection in this population. The findings strongly support the idea that gastric cancer is primarily determined by environmental factors and develops in a multistep progression of precancerous lesions. PMID- 16246222 TI - Common single nucleotide polymorphism of hypoxia-inducible factor-1alpha and its impact on the clinicopathological features of esophageal squamous cell carcinoma. AB - OBJECTIVE: Angiogenesis is one of the most important molecular events in solid tumor development and growth, in which hypoxia-inducible factor (HIF)-1alpha is a key regulator and plays an important role. Studies have shown that a single nucleotide polymorphism (C1772T) in the HIF-1alpha gene exerts a large effect on the phenotype of human head and neck squamous cell carcinoma and renal cell carcinoma. But the impact of the C1772T polymorphism on the clinicopathological features of human esophageal squamous cell carcinoma (ESCC) remains unknown, and thus it is the main focus of our study. METHODS: The C1772T genotype of 95 ESCC patients and 104 healthy controls were studied by using the polymerase chain reaction and restriction fragment length polymorphism. Mutations were confirmed by direct DNA sequencing. The impact of C1772T on tumor size, invasive depth, lymph node metastasis, distant metastasis, histological grade and TNM stage was also studied. RESULTS: The genotype frequency observed in the patients and controls was 11.58% versus 10.58%, respectively, for genotype C/T (P > 0.05). Genotype T/T was not found in our study. Larger tumors and a higher rate of lymph node metastasis was found for the C/T group. CONCLUSIONS: Although there is no significant difference of genotype distribution between ESCC patients and healthy controls, genotype C/T is associated with larger tumor and higher rate of lymph node metastasis. PMID- 16246223 TI - Preoperative diagnosis of gastric cancer using 2-D magnetic resonance imaging with 3-D reconstruction techniques. AB - OBJECTIVE: To investigate the clinical value of 2-D magnetic resonance imaging (MRI) with 3-D reconstruction techniques for the preoperative diagnosis and TNM staging of gastric cancer. METHODS: Using a Philips Gyroscan NT 1.0T superconductive unit, MRI using the water-filling method was performed in 15 patients with suspected gastric cancers. The 2-D MRI sequences included TSE-T1WI, TSE-T2WI and fat suppression (SPIR). The source images of magnetic resonance hydrography (heavily TSE-T2WI sequence) were reconstructed using the Philips EasyVision viewing workstation. Four 3-D postprocessing algorithms, including maximum intensity projection, surface shaded viewing, volume rendering and virtual endoscopy, were performed and compared with the results of a barium study and endoscopy. All 15 patients with 16 gastric cancers had their diagnosis confirmed by postoperative pathological findings. RESULTS: 2-D MRI and 3-D reconstruction images were successfully obtained for all 15 patients. The maximum intensity projection, surface shaded viewing, and volume rendering images corresponded to the upper gastrointestinal series findings, and the virtual endoscopy images corresponded to the gastroscopic views. In 16 gastric lesions, MRI correctly diagnosed 14 (87.5%) advanced gastric cancers, and the tumor location, size and classification were also accurately identified. The accuracy of MRI for determining the preoperative TNM stage was 64.3% (9/14), and there was significant correlation between these results and those from the histopathological studies (P < 0.01). Based on T, N and M factors, the staging accuracy of MRI was 71.4% (10/14), 57.1% (8/14) and 85.7% (12/14), respectively. CONCLUSIONS: 2-D MRI with 3-D reconstruction is an effective method for the preoperative diagnosis and TNM staging of gastric cancer. However, the detection of early cancers or benign lesions and N-staging should be further studied. PMID- 16246224 TI - Laparoscopic versus open appendectomy in West Bengal, India. AB - OBJECTIVE: Laparoscopic appendectomy, although widely practiced, has not gained universal approval. Laparoscopic appendectomy in India is relatively new, and reports regarding its use are few. The aim of this study was to compare laparoscopic with open appendectomy and ascertain its therapeutic benefit, if any, in the overall management of acute appendicitis. METHODS: The study group consisted of 279 patients suffering from acute appendicitis. One hundred patients underwent laparoscopic appendectomy (LA) and 179 patients underwent open appendectomy (OA). Comparisons were based on median values for length of hospital stay, operating time, postoperative morbidity, duration of convalescence and operative cost. Mann-Whitney statistics (T) were calculated and, because of the large sample size, the normal deviate test (Z) was used. RESULTS: Of the 100 patients who were to receive LA, six patients (6%) had the procedure converted to open surgery. The rate of overall complications (LA 15%vs OA 31.8%; P < 0.001) was significantly lower in patients undergoing LA. The median length of stay was significantly shorter after LA (3 days after LA vs 5 days after OA; P < 0.0001) than after OA. The operating time was shorter (median values: OA 25 min vs LA 28 min; 0.01 < P < 0.05) in patients undergoing OA compared with LA. CONCLUSION: For LA, hospital stay was significantly shorter, and the one-time operative cost appeared to be marginally the same as open surgery. Furthermore, LA was associated with increased clinical comfort in terms of fewer wound infections, faster recovery, earlier return to work and improved cosmesis. PMID- 16246225 TI - Correlation between a gene polymorphism of tumor necrosis factor and inflammatory bowel disease. AB - OBJECTIVES: To analyze polymorphism of the tumor necrosis factor (TNF) gene in inflammatory bowel disease (IBD) patients from the Han Chinese ethnic group, and to investigate the role of polymorphism in the pathogenesis of IBD. METHODS: Polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) techniques were used to analyze gene polymorphisms in the TNF-alpha and TNF-beta genes in 131 patients with IBD. RESULTS: The genotype frequency and allelic frequency of TNF-alpha-308 in patients with ulcerative colitis (UC) were 15.5% and 8.7%, respectively, significantly higher than the control population (4.1% and 2.0%, respectively; P < 0.001). There was no significant difference between patients with Crohn's disease (CD) and the normal population with regard to the genotype frequency and allelic frequency of TNF-alpha-308, and neither were there any differences with regard to TNF-beta+252 in patients with IBD (UC and CD) and the normal population. The TNF-alpha-308 polymorphism and the TNF beta+252 loci did not correlate with age, gender, disease activity or lesion site for IBD patients. CONCLUSIONS: The TNF-alpha-308 allele may be related to susceptibility to UC. The TNF-alpha-308 gene polymorphism is not involved in pathogenesis of CD. No correlation was found between the TNF-beta+252 polymorphism and IBD. Polymorphisms of the TNF-alpha-308 and TNF-beta+252 loci do not correlate with age, gender, disease activity or lesion site. PMID- 16246226 TI - Crohn's disease in mainland China: a systematic analysis of 50 years of research. AB - BACKGROUND: Crohn's disease appears to be increasing in frequency in many areas of the world. However, little information with regard to disease incidence, prevalence and temporal trends has been published in China. OBJECTIVE: The aim of this review is to better understand the occurrence of Crohn's disease in mainland China, and to give an overview of the current status of the disease. METHODS: We used a computer-based literature search using 50-years of records from the Chinese Database of Biology and Medicine (CBM, 1979-2002), combined with a manual year-by-year search of the literature from 1950 to 1979. Each article was double reviewed, and all descriptive epidemiological data were recorded, pooled and statistically analyzed. RESULTS: In total, 1526 cases of Crohn's disease since 1950 have been reported by more than 50 hospitals from 22 provinces and cities in mainland China, comprising 884 male and 642 female patients, with a 1.38:1 male predominance. More than 70% of patients were aged 20 to 50 years old, with a mean age of 37.2 +/- 2.68 (ranging from 1 to 83) years. The extrapolated disease incidence and prevalence rates are 0.28/100,000 person years and 1.38/100,000 persons, respectively. CONCLUSIONS: The incidence and prevalence rates of Crohn's disease are still lower than those in Western and other Asian countries, but these rates have been increasing rapidly, and the disease is no longer uncommon. An underestimation may occur because the patients who were misdiagnosed or did not seek medical advice could not be included in the study. A future population based survey is warranted. PMID- 16246227 TI - Epidemiology and genotypes of HEV in Wuhan. AB - BACKGROUND: Understanding the genotype and clinical features of the hepatitis E virus (HEV) are important for understanding its characteristics, for evaluating region-specific diagnostic assays, and producing vaccines. OBJECTIVES: To investigate the epidemiology and the genotypes of HEV among outpatients and inpatients in the Department of Infectious Diseases of Tongji Hospital in Wuhan, China. METHODS: Clinical data were elicited from the hospital records of patients who were clinically diagnosed with acute hepatitis between January 2000 and August 2004 (4920 patients). Of these cases, 120 patients with anti-HEV-IgM, IgG positive were selected to analysis. Conserved genomic sequences of open reading frame 2 (345 bp) in the HEV gene were detected using polymerase chain reaction, 25 of which were cloned and sequenced. Clustal X and Mega software were used for phylogenetic analysis of genotypes strains. RESULTS: The HEV infection rate is gradually increasing in Wuhan. The number of male patients was 3.3-fold greater than the number of female patients found in clinical investigations. People aged 30-59 years are more susceptible to infection, and people are more susceptible in March-June. Twenty-five isolates shared the same genotype, genotype IV, with 82.61-98.55% nucleotide identity. This genotype had 76.52-81.74%, 70.43-73.04%, 76.52-81.16%, and 84.35-88.70% homology with the nucleotide sequence of HEV genotypes I-IV, respectively. Phylogenetic analysis suggested that these 25 isolates represented at least three different subtypes, but there were no significant differences found in the epidemiological features or liver function of patients with the three subtypes. CONCLUSIONS: HEV sequences isolated from patients in Wuhan belong to different subtypes of HEV genotype IV. PMID- 16246228 TI - Effects of sodium phenylbutyrate on differentiation and induction of the P21WAF1/CIP1 anti-oncogene in human liver carcinoma cell lines. AB - OBJECTIVES: To explore the effects of sodium phenylbutyrate on the proliferation, differentiation, cell cycle arrest and induction of the P(21WAF1/CIP1) anti oncogene in human liver carcinoma cell lines Bel-7402 and HepG2. METHODS: Bel 7402 and HepG2 human liver carcinoma cells were treated with sodium phenylbutyrate at different concentrations. Light microscopy was used to observe morphological changes in the carcinoma cells. Effects on the cell cycle were detected by using flow cytometry. P(21WAF1/CIP1) expression was determined by both reverse transcription-polymerase chain reaction and western blotting. Statistical analysis was performed by using one-way anova and Student's t-test. RESULTS: Sodium phenylbutyrate treatment caused time- and dose-dependent growth inhibition of Bel-7402 and HepG2 cells. This treatment also caused a decline in the proportion of S-phase cells and an increase in the proportion of G(0)/G(1) cells. Sodium phenylbutyrate increased the expression of P(21WAF1/CIP1). CONCLUSIONS: Sodium phenylbutyrate inhibits the proliferation of human liver carcinoma cells Bel-7402 and HepG2, induces partial differentiation, and increases the expression of P(21WAF1/CIP1). PMID- 16246230 TI - Guidelines for the diagnosis and treatment of chronic pancreatitis (Nanjing, 2005). PMID- 16246229 TI - Experimental acute alcohol pancreatitis-related liver damage and endotoxemia: synbiotics but not metronidazole have a protective effect. AB - OBJECTIVE: The aim of this study was to test the effect of gut manipulation by either novel synbiotics or by metronidazole on either endotoxemia or the severity of liver damage in the course of acute pancreatitis from alcohol ingestion. METHODS: Sprague-Dawley rats were fed for 1 week through an intragastric tube a liquid diet with either: (i) 1 mL t.i.d. of a mixture of synbiotics (Lactobacillus acidophilus, Lactobacillus helveticus and Bifidobacterium in an enriched medium); (ii) 20 mg/kg t.i.d. metronidazole; or (iii) standard diet. Then, acute pancreatitis was induced by caerulein and when the disease was full blown, rats were fed an alcohol-rich diet. Synbiotic and metronidazole treatment was given for a further 2 weeks. Transaminase and endotoxemia levels were measured before treatment, after 6 h, after 24 h and 2 weeks later, at the time the rats were killed. Liver samples were obtained for histological analysis. RESULTS: Synbiotics but not metronidazole improved the acute pancreatitis-induced increase in endotoxemia and transaminase levels. The addition of alcohol worsened these variables to a limited extent in the synbiotic-treated group, while metronidazole had a negative effect on liver damage. CONCLUSIONS: Gut flora pretreatment with synbiotics was able to effectively protect against endotoxin/bacterial translocation, as well as liver damage in the course of acute pancreatitis and concomitant heavy alcohol consumption. The beneficial effect of synbiotics on liver histology seems to be correlated with endotoxemia. Metronidazole did not produce such a beneficial effect; in fact, it further worsened liver damage when alcohol was added to the background of ongoing acute pancreatic inflammation. PMID- 16246231 TI - Comparison of the efficacy of 1-day high-dose quadruple therapy versus 7-day triple therapy for treatment of Helicobacter pylori infection. AB - BACKGROUND: The proton pump inhibitor (PPI)-based 7-day triple therapy is the regimen with the highest cure rates for eradication of Helicobacter pylori infection and has been recommended as the first-line regimen in the world. It had been reported that a 1-day quadruple therapy could also successfully cure 95% of the H. pylori infected patients. OBJECTIVES: To observe the efficacy of 1-day high-dose quadruple therapy versus 7-day triple therapy for treatment of H. pylori infection, and to observe side-effects of the two different regimens. METHODS: This randomized, open, parallel-controlled study was conducted at Renji Hospital between November 2004 to March 2005. A total of 80 consecutive patients with non-ulcer dyspepsia, who were H. pylori positive proven by both rapid urease test and histology were included and randomly assigned to 1-day quadruple therapy or 7-day triple therapy. Thirty-nine patients were administered with 1-day high dose quadruple therapy including esomeprazole 40 mg b.i.d., colloidal bismuth subcitrate 440 mg q.i.d., amoxicillin 2 g q.i.d. and metronidazole (400 mg q.i.d.) for 1 day. Forty-one patients received a standard 7-day triple therapy consisting of esomeprazole 20 mg b.i.d., clarithromycin 500 mg b.i.d. and amoxicillin 1 g b.i.d. for 7 days. The eradication rates were evaluated by the (13)C-urea breath test at least 4 weeks after completion of a course treatment. RESULTS: Seventy-seven patients completed the trial and three patients dropped out. The eradication rates in the 1-day therapeutic group and the 7-day therapeutic group were 39.5% (15/38) and 84.6% (33/39), respectively. There was a statistically significant difference between the two groups (P < 0.0001). Short lasting and self-limiting side effects including thirst, a metallic taste, diarrhea and abdominal pain were reported in three patients (7.9%) in the 1-day group and seven patients (18%) in the 7-day group (P = 0.31). CONCLUSIONS: A 1 day high-dose quadruple therapy with amoxicillin, metronidazole, bismuth salt, and esomeprazole is not effective for eradication of H. pylori compared with the standard 7-day triple therapy. PMID- 16246232 TI - Hypereosinophilic syndrome presenting with eosinophilic colitis, enteritis and cystitis. PMID- 16246234 TI - The learning brain: lessons for education: a precis. PMID- 16246233 TI - Three-month-olds, but not newborns, prefer own-race faces. AB - Adults are sensitive to the physical differences that define ethnic groups. However, the age at which we become sensitive to ethnic differences is currently unclear. Our study aimed to clarify this by testing newborns and young infants for sensitivity to ethnicity using a visual preference (VP) paradigm. While newborn infants demonstrated no spontaneous preference for faces from either their own- or other-ethnic groups, 3-month-old infants demonstrated a significant preference for faces from their own-ethnic group. These results suggest that preferential selectivity based on ethnic differences is not present in the first days of life, but is learned within the first 3 months of life. The findings imply that adults' perceptions of ethnic differences are learned and derived from differences in exposure to own- versus other-race faces during early development. PMID- 16246235 TI - Paving the way towards meaningful interactions between neuroscience and education. PMID- 16246236 TI - The brain in the classroom? The state of the art. PMID- 16246237 TI - An invaluable foundation for better bridges. PMID- 16246238 TI - Young children's rapid learning about artifacts. AB - Tool use is central to interdisciplinary debates about the evolution and distinctiveness of human intelligence, yet little is actually known about how human conceptions of artifacts develop. Results across these two studies show that even 2-year-olds approach artifacts in ways distinct from captive tool-using monkeys. Contrary to adult intuition, children do not treat all objects with appropriate properties as equally good means to an end. Instead, they use social information to rapidly form enduring artifact categories. After only one exposure to an artifact's functional use, children will construe the tool as 'for' that particular purpose and, furthermore, avoid using it for another feasible purpose. This teleo-functional tendency to categorize tools by intentional use represents a precursor to the design stance - the adult-like tendency to understand objects in terms of intended function - and provides an early foundation for apparently distinctive human abilities in efficient long-term tool use and design. PMID- 16246239 TI - Foundations for self and other: a study in autism. AB - There is controversy over the basis for young children's experience of themselves and other people as separate yet related individuals, each with a mental perspective on the world - and over the nature of corresponding deficits in autism. Here we tested a form of self-other connectedness (identification) in children with and without autism, who were group-matched according to CA (approximately 6 to 16 years) and verbal MA (approximately 2 1/2 to 14 years), and therefore IQ. We gave two forms of a novel 'sticker test' in which children needed to communicate to another person where on her body she should place her sticker-badge. Across the trials of Study 1, all of the non-autistic children pointed to their own bodies at least once, but over half the children with autism failed to point to themselves at all, even though they communicated successfully in other ways. In Study 2, where a screen was introduced to hide the tester's body, group differences in the children's communicative self-orientated gestures were most marked after the tester had 'modelled' a point-to-herself gesture in communicating to the child. Our interpretation is that autism involves a relative failure to adopt the bodily-anchored psychological and communicative stance of another person. We suggest that this process of identification is essential to self-other relations and grounds young children's developing understanding of minds. PMID- 16246240 TI - One-year-olds comprehend the communicative intentions behind gestures in a hiding game. AB - This study explored infants' ability to infer communicative intent as expressed in non-linguistic gestures. Sixty children aged 14, 18 and 24 months participated. In the context of a hiding game, an adult indicated for the child the location of a hidden toy by giving a communicative cue: either pointing or ostensive gazing toward the container containing the toy. To succeed in this task children had to do more than just follow the point or gaze to the target container. They also had to infer that the adult's behaviour was relevant to the situation at hand - she wanted to inform them that the toy was inside the container toward which she gestured. Children at all three ages successfully used both types of cues. We conclude that infants as young as 14 months of age can, in some situations, interpret an adult behaviour as a relevant communicative act done for them. PMID- 16246241 TI - Fast mapping between a phrasal form and meaning. AB - This is the first study to investigate experimentally how children come to learn mappings between novel phrasal forms and novel meanings: a central task in learning a language. Two experiments are reported. In both studies 5- to 7-year old children watched a short set of video clips depicting objects appearing in various ways. Each scene was described using a novel verb embedded in a novel construction. Children who watched the videos and heard the accompanying description were able to match new descriptions that used the novel construction with new scenes of appearance. Moreover, our results suggest a facilitative effect for the disproportionately high frequency of occurrence of a single verb in a particular construction (such as has been found to exist in naturalistic input to children). While the fast mapping might be taken as an indication of innate knowledge that is specific to language, analogous effects in non linguistic categorization tasks suggest that children are acquiring the new phrasal form with general cognitive skills. PMID- 16246242 TI - Sensitivity to social contingencies between 1 and 3 months of age. AB - Infants' sensitivity to social contingencies was assessed. In Study 1, 1-month old infants and their mothers interacted face-to-face in three types of imperfect contingent interactions: Normal, Non-Contingent and Imitation. One-month-old infants did not discriminate these conditions. In Study 2, 3-month-old infants were tested as in Study 1. At 3 months of age, infants gazed reliably longer in the Imitation condition and smiled reliably more in the Normal than in the Non Contingent and Imitation interactions. These findings suggest a developmental transition in the sensitivity to social contingencies between 1 and 3 months of age. The relationship between the developing sensitivity to social contingencies and social cognition is discussed. PMID- 16246243 TI - How do parents respond to children's questions about the identity of artifacts? AB - We investigated how parents respond to young children's questions about the identity of artifacts. Children's questions were predominantly ambiguous about whether they were inquiring about name or function, but when their questions were more specific, they were almost always about function. For unfamiliar objects, parents responded with functional information the majority of the time, alone or in addition to names. For atypical members of familiar categories, adults usually responded only with the category name. The results indicate that adults adjust their responses in a way that often provides the information about object kind, specifically functional information in the case of artifacts, that they believe their children are lacking. Such input may contribute to the development of children's concepts and word meanings. PMID- 16246244 TI - Maturation of action monitoring from adolescence to adulthood: an ERP study. AB - This study investigated the development of the frontal lobe action-monitoring system from late childhood and adolescence to early adulthood using ERP markers of error processing. Error negativity (ERN) and correct response negativity (CRN) potentials were recorded while adolescents and adults (aged 12-22 years, n = 23) performed two forced-choice visual reaction time tasks of differing complexity. Significant age differences were seen for behavioural and ERP responses to complex (infrequent, incompatible) trials: adolescents elicited an error negativity of reduced magnitude compared with adults. Furthermore, in contrast to adults, adolescents showed a non-significant differentiation between response locked ERP components elicited by correct (CRN) and error responses (ERN). Behaviourally, adolescents corrected fewer errors in incompatible trials, and with increasing age there was greater post-error slowing. In conclusion, the neural systems underlying action-monitoring continue to mature throughout the second decade of life, and are associated with increased efficiency for fast error detection and correction during complex tasks. PMID- 16246246 TI - Beyond the distributional input? A developmental investigation of asymmetry in infants' categorization of cats and dogs. AB - Two experiments are reported using a visual familiarization categorization procedure. In both experiments, infants were familiarized with sets of stimuli previously shown to contain asymmetric feature distributions that support an asymmetry in young infants' categorization of cats and dogs (i.e. infants' cat category excludes dogs but their dog category includes cats). In Experiment 1, the asymmetry was replicated in 4-month-old infants. In contrast, 10-month-old infants demonstrated exclusive category representations for both cats and dogs. In Experiment 2, an additional group of 10-month-olds demonstrated exclusive representations for both cats and dogs under conditions of very limited within task category familiarization. Potential mechanisms underlying the shift from an asymmetric to a symmetric pattern of categorization in the first year are discussed. PMID- 16246245 TI - The development of gaze following and its relation to language. AB - We examined the ontogeny of gaze following by testing infants at 9, 10 and 11 months of age. Infants (N = 96) watched as an adult turned her head toward a target with either open or closed eyes. The 10- and 11-month-olds followed adult turns significantly more often in the open-eyes than the closed-eyes condition, but the 9-month-olds did not respond differentially. Although 9-month-olds may view others as 'body orienters', older infants begin to register whether others are 'visually connected' to the external world and, hence, understand adult looking in a new way. Results also showed a strong positive correlation between gaze-following behavior at 10-11 months and subsequent language scores at 18 months. Implications for social cognition are discussed in light of the developmental shift in gaze following between 9 and 11 months of age. PMID- 16246247 TI - Children's implicit knowledge of harmony in Western music. AB - Three experiments examined children's knowledge of harmony in Western music. The children heard a series of chords followed by a final, target chord. In Experiment 1, French 6- and 11-year-olds judged whether the target was sung with the vowel /i/ or /u/. In Experiment 2, Australian 8- and 11-year-olds judged whether the target was played on a piano or a trumpet. In Experiment 3, Canadian 8- and 11-year-olds judged whether the target sounded good (i.e. consonant) or bad (dissonant). The target was either the most stable chord in the established musical key (i.e. the tonic, based on do, the first note of the scale) or a less stable chord. Performance was faster (Experiments 1, 2 and 3) and more accurate (Experiment 3) when the target was the tonic chord. The findings confirm that children have implicit knowledge of syntactic functions that typify Western harmony. PMID- 16246248 TI - Evidence against a maximum response model of exogenous visual orienting during early infancy and support for a dimensional switching model. AB - Very young infants orient overtly with eye and head movements to salient events in their visual environments, but those events rarely occur in the absence of competing visual stimuli. Two different models of how this kind of orienting is related to number and distribution of elements in the stimulus field were tested with infants across the age range from 2 to 5 months in four experiments. A set size manipulation in Experiments 1-3 produced data that were mostly inconsistent with the Maximum Response model proposed by Dannemiller (1998), especially at ages over 3 months. Experiment 4 produced data from 3.5-month-olds that were consistent with an alternative Dimensional Switching model that assumed that there was switching across trials in the stimulus dimension that drove orienting. This Dimensional Switching model can explain the small to nonexistent set size effects observed in the first three experiments as well as data from previous experiments using this paradigm. Factors that could produce this kind of dimensional switching over time were considered and other implications of this model for understanding the development of overt visual orienting were discussed. PMID- 16246249 TI - What young children do and do not know about the spelling of inflections and derivations. AB - Morphemes have a powerful impact on the spellings of words in English. We report on two experiments examining young children's knowledge of the effect of suffix morphemes on spelling. In Experiment 1, 5- to 8-year-olds demonstrated awareness of the role of inflections, but not derivations in spelling. Experiment 2 examined whether children might show an understanding of the impact of derivations on spelling when they are provided with the specific spelling pattern. There was support for this idea; 7- to 10-year-old children took advantage of the derivations that were given to spell other derivations, but not the endings of control words. The results of these experiments suggest that young children are aware of the role that suffixes play in determining spelling, but that there are limits, such as those imposed by spelling difficulty, that constrain the execution of this knowledge. These findings provide new insights into current theoretical debates on spelling development. PMID- 16246250 TI - Ambiguous benefits: the effect of bilingualism on reversing ambiguous figures. AB - Two studies are reported in which monolingual and bilingual children, approximately 6 years old, attempted to identify the alternative image in a reversible figure. In both studies, bilingual children were more successful than monolinguals in seeing the other meaning in the images. In the first study, there was no relation between the ability to reverse the interpretation and performance on the children's embedded figures task, a task that superficially appeared to involve similar processes. The second study replicated this finding but showed that performance was strongly related to success in the post-switch phase of the dimensional change card sort task. In both cases, the meaning of an image must be reassigned, and bilinguals were better in both these tasks. PMID- 16246251 TI - A longitudinal investigation of visual event-related potentials in the first year of life. AB - The goal of the current study was to assess general maturational changes in the ERP in the same sample of infants from 4 to 12 months of age. All participants were tested in two experimental manipulations at each age: a test of facial recognition and one of object recognition. Two sets of analyses were undertaken. First, growth curve modeling with mixed models was used to examine trajectories of development and possible differences in trajectories based on recognition memory (novel versus familiar) and/or stimulus-specific memory (face versus object recognition). Our results suggest that the Pb, Nc and Slow Wave components change significantly in terms of amplitude and latency over the first year of life. Pb amplitude showed a significant non-linear increase over time, whereas Pb latency showed a significant linear decrease over time with a plateau beginning at 10 months. Nc amplitude showed a significant linear decrease over time (i.e. a stronger negative value), whereas Nc latency showed a significant linear decrease over time, with a plateau beginning at 8 months. Second, to relate our findings to those reported in the literature, we examined the effects of memory and stimulus and their combination. Differences between recognition memory and stimulus specific memory were found in the responses to familiar and novel faces and objects for all three components, although the pattern differed across the five ages. These results have implications for future studies that involve the recording of the visual ERP, and point to the advantages of growth curve modeling in examining longitudinal data to account for non-linear development. PMID- 16246252 TI - Expression studies on a novel type 2B variant of the von Willebrand factor gene (R1308L) characterized by defective collagen binding. AB - A novel mutation, R1308L (3923G > T) was present in the heterozygous state in five members of a family with type 2B von Willebrand disease (VWD) characterized by a full set of von Willebrand factor (VWF) multimers in plasma and by the absence of thrombocytopenia before and after desmopressin (DDAVP). The defect (R1308L) was located at the same amino acid position of one of the most common mutations associated with type 2B VWD (R1308C), which is characterized by the loss of high molecular weight VWF multimers (HMWM) in plasma and the occurrence of thrombocytopenia. To understand the mechanisms of this defect, the novel (R1308L) and 'common' (R1308C) mutations were expressed in COS-7 cells, either alone or, to mimic the patients' heterozygous state, together with wild-type VWF. R1308L recombinant VWF (rVWF) had a higher affinity for the platelet glycoprotein Ibalpha (GPIbalpha) receptor than wild-type rVWF, R1308C rVWF showing an even higher affinity. A novel finding was that both mutant rVWFs showed a similarly reduced binding to collagen type I and type III in comparison with wild-type rVWF. The latter finding suggests a more important role than recognized so far for the VWF A1 domain in VWF binding to collagen, which may contribute to the in vivo hemostatic defect associated with type 2B VWD. PMID- 16246253 TI - Chemically modified thrombin and anhydrothrombin that differentiate macromolecular substrates of thrombin. AB - BACKGROUND: Thrombin is a primary inducer of thrombus formation by activations of coagulation cascade and platelet aggregation. Hitherto, several types of thrombin inhibitors have been developed for therapeutic purpose. OBJECTIVES: We prepared modified thrombin (M-thrombin) and modified anhydrothrombin (M-anhydrothrombin) by chemical modification of carboxyl groups of thrombin and anhydrothrombin, respectively, to present a new strategy for a potent antiplatelet-anticoagulant agent and new tools for investigation of thrombin functions. RESULTS: M anhydrothrombin retained high affinity for factor VIII (FVIII), but demonstrated lower affinity than anhydrothrombin for fibrinogen and factor V (FV). Both M anhydrothrombin and anhydrothrombin prolonged activated partial thromboplastin time (APTT) without affecting prothrombin time, and M-anhydrothrombin prolonged APTT much more than anhydrothrombin. M-anhydrothrombin also retained affinity for the recombinant extracellular domain peptide of protease-activated receptor 1 (PAR1). M-thrombin exhibited marginal clotting activity (4% of thrombin), but induced platelet aggregation in platelet-rich plasma without forming a fibrin clot, which was completely suppressed by anti-PAR1 antibody (ATAP2) and by M anhydrothrombin, but not by anhydrothrombin. These results indicate that M thrombin induced platelet aggregation through the activation of PAR1, and M anhydrothrombin inhibited this process completely. In contrast, neither M anhydrothrombin nor anhydrothrombin apparently inhibited thrombin-induced platelet aggregation. Only in the presence of the Gly-Pro-Arg-Pro (GPRP) peptide that inhibits polymerization of fibrin, M-anhydrothrombin completely inhibited thrombin-induced platelet aggregation. CONCLUSION: M-thrombin is PAR1-specific and M-anhydrothrombin is FVIII- and PAR1-specific derivatives, and thereby, are new tools as specific agonist and antagonist, respectively, of PAR1. Furthermore, M-anhydrothrombin may be an attractive model for development of a potent anticoagulant-antiplatelet agent. PMID- 16246254 TI - Comparison of cell-surface TFPIalpha and beta. AB - BACKGROUND: Tissue factor pathway inhibitor (TFPI) is mainly produced by endothelial cells and alternative mRNA splicing generates two forms, TFPIalpha and TFPIbeta. A portion of expressed TFPI remains associated with the cell surface through both direct (TFPIbeta) and indirect (TFPIalpha) glycosylphosphatidyl-inositol (GPT)-mediated anchorage. OBJECTIVE: Compare the structure and properties of TFPIalpha and TFPIbeta. METHODS: TFPIalpha and TFPIbeta, with protein molecular masses of 36 and 28 kDa, respectively, migrate similarly (46 kDa) on SDS-PAGE. Experiments using specific glycosidases were carried out to determine the different glycosylation pattern of the two forms. ECV304 cells, a cell line with some endothelial properties, were stimulated with IL-lbeta, LPS, and TNFalpha for up to 24 hrs and mRNA levels and protein synthesis were determined. Stable clones of ECV304 cells that express reduced levels of TFPIalpha, TFPIbeta or both were produced using a plasmid-based small interfering RNA technique. Surface TFPI activity was determined by a two-stage chromogenic assay based on the ability of each form to inhibit FXa activation by FVIIa on cells with comparable amount of tissue factor (TF). RESULTS AND CONCLUSIONS: The deglycosylation studies show that the difference in molecular masses is due to a greater degree of sialylation in O-linked carbohydrate in TFPIbeta. The mRNA and protein levels of neither form of TFPI were affected by stimulation of cells with inflammatory stimuli. Although TFPIalpha comprises 80% of the surface-TFPI, TFPIbeta was responsible for the bulk of the cellular FVIIa/TF inhibitory activity, suggesting a potential alternative role for cell surface TFPIalpha. PMID- 16246256 TI - Inherited defects of coagulation Factor V: the thrombotic side. AB - DNA variations in the Factor V gene have played a major role in thrombosis research ever since the discovery of Factor V Leiden. Here, all relatively common DNA variations in the coding regions of the Factor V gene are discussed. Many of them have been associated with venous thrombosis or related diseases. However, most variations have been studied separately, without taking the presence of other variations in the same gene into account. This means that their association with disease should be interpreted with caution, as it may reflect linkage with another variation. An approach in which a haplotype-based analysis of the Factor V gene is combined with in vitro assays of recombinant proteins is advocated. Finally, a possible reason for the relatively polymorphic nature of the Factor V protein is discussed. PMID- 16246255 TI - Anti-inflammatory effect of activated protein C in gastric epithelial cells. AB - It has been previously demonstrated that activated protein C (APC) plays an important role in the inhibition of inflammation in the gastric mucosa from patients with Helicobacter pylori infection. However, the role of gastric epithelial cells in the anti-inflammatory activity of APC remains unknown. In the present study, we evaluated the anti-inflammatory activity of APC and the expression of thrombomodulin (TM) and endothelial protein C receptor (EPCR) in gastric epithelial cells. The gastric epithelial cell lines, MKN-1 and AGS, and gastric biopsy samples from patients with and without H. pylori infection were used in the experiments. Polymerase chain reaction showed that gastric epithelial cell lines express EPCR and TM. Flow cytometry analysis also showed EPCR expression in both cells. H. pylori infection significantly increased EPCR expression compared with non-infected cells. Similar results were observed in vivo when samples from patients with and without H. pylori infection were analyzed for EPCR protein expression. Significant TM activity was found on AGS and MKN-1 cells stimulated with LPS from Escherichia coli and VacA antigen. APC was able to significantly inhibit the secretion of MCP-1 and IL-1beta induced by H. pylori homogenate in AGS cells. APC also remarkably suppressed the mRNA expression and secretion of MCP-1 from AGS cells infected with H. pylori. These results demonstrated the expression of components of the protein C pathway on gastric epithelial cells and that APC may play a critical role in the protection against gastric mucosal inflammation. PMID- 16246257 TI - Expression of POU-domain transcription factor, Oct-6, in schizophrenia, bipolar disorder and major depression. AB - BACKGROUND: The POU-domain transcription factor Oct-6 has been reported to be differentially expressed between schizophrenic and control post-mortem brains. In this study, we attempted to replicate this finding and to discover whether Oct-6 was also dysregulated in bipolar disorder and major depression. METHODS: Oct-6 mRNA and protein expression were determined by in-situ hybridization and immunohistochemistry respectively in sections of post-mortem brain. RESULTS: We did not observe any differences in Oct-6 expression between any of the groups under study. Oct-6 mRNA and protein was identically expressed in the hippocampal and cortical regions of most specimens in all groups, including controls. CONCLUSION: Oct-6 is, therefore, unlikely to be a specific marker for any psychological disorder; rather its expression in controls suggests that it is normally expressed in most adult brains. PMID- 16246258 TI - Heat shock protein and heat shock factor 1 expression and localization in vaccinia virus infected human monocyte derived macrophages. AB - BACKGROUND: Viruses remain one of the inducers of the stress response in the infected cells. Heat shock response induced by vaccinia virus (VV) infection was studied in vitro in human blood monocyte derived macrophages (MDMs) as blood cells usually constitute the primary site of the infection. METHODS: Human blood monocytes were cultured for 12-14 days. The transcripts of heat shock factor 1 (HSF1), heat shock protein 70 (HSP70), heat shock protein 90 (HSP90) and two viral genes (E3L and F17R) were assayed by reverse transcriptase-polymerase chain reaction (RT-PCR), and the corresponding proteins measured by Western blot. Heat shock factor 1 DNA binding activities were estimated by electrophoretic mobility shift assay (EMSA) and its subcellular localization analyzed by immunocytofluorescence. RESULTS: It appeared that infection with vaccinia virus leads to activation of the heat shock factor 1. Activation of HSF1 causes increased synthesis of an inducible form of the HSP70 both at the mRNA and the protein level. Although HSP90 mRNA was enhanced in vaccinia virus infected cells, the HSP90 protein content remained unchanged. At the time of maximum vaccinia virus gene expression, an inhibitory effect of the infection on the heat shock protein and the heat shock factor 1 was most pronounced. Moreover, at the early phase of the infection translocation of HSP70 and HSP90 from the cytoplasm to the nucleus of the infected cells was observed. CONCLUSION: Preferential nuclear accumulation of HSP70, the major stress-inducible chaperone protein, suggests that VV employs this particular mechanism of cytoprotection to protect the infected cell rather than to help viral replication. The results taken together with our previous data on monocytes or MDMs infected with VV or S. aureus strongly argue that VV employs multiple cellular antiapoptotic/cytoprotective mechanisms to prolong viability and proinflammatory activity of the cells of monocytic-macrophage lineage. PMID- 16246259 TI - Alcohol reversibly disrupts TNF-alpha/TACE interactions in the cell membrane. AB - BACKGROUND: Alcohol abuse has long been known to adversely affect innate and adaptive immune responses and pre-dispose to infections. One cellular mechanism responsible for this effect is alcohol-induced suppression of TNF-alpha (TNF) by mononuclear phagocytes. We have previously shown that alcohol in part inhibits TNF-alpha processing by TNF converting enzyme (TACE) in human monocytes. We hypothesized that the chain length of the alcohol is critical for post transcriptional suppression of TNF secretion. METHODS: Due to the complex transcriptional and post-transcriptional regulation of TNF in macrophages, to specifically study TNF processing at the cell membrane we performed transient transfections of A549 cells with the TNF cDNA driven by the heterologous CMV promoter. TNF/TACE interactions at the cell surface were assessed using fluorescent resonance energy transfer (FRET) microscopy. RESULTS: The single carbon alcohol, methanol suppressed neither TNF secretion nor FRET efficiency between TNF and TACE. However, 2, 3, and 4 carbon alcohols were potent suppressors of TNF processing and FRET efficiency. The effect of ethanol, a 2 carbon alcohol was reversible. CONCLUSION: These data show that inhibition of TNF alpha processing by acute ethanol is a direct affect of ethanol on the cell membrane and is reversible upon cessation or metabolism. PMID- 16246261 TI - The Self Test: a screening tool for dementia requiring minimal supervision. AB - BACKGROUND: The main purpose of this study was to correlate a newly developed, simple, but comprehensive screening test for dementia called the Self Test (ST) with the Mini-mental State Examination (MMSE), and establish the reliability and validity of the ST. METHODS: The ST was administered to 42 consecutive new referrals to the memory clinic at the Cole Neuroscience Center, University of Tennessee Medical Center (UTMC), Knoxville, TN, U.S.A. and 41 age-matched control subjects at regular intervals over a 12-month period. RESULTS: The total ST scores correlated well with cognitive severity as measured by the MMSE (r = 0.71, p < 0.01). The area under the receiver-operating characteristic curve (AUROCC) in the group of patients with Alzheimer's disease (AD) with less than a high school education was 0.80, and 0.89 in those with at least a high school education. Reliability analysis showed a mean inter-item correlation of 0.31 for patients with AD, and 0.47 for normal participants. Cronbach's alpha was calculated to be 0.70 for the AD group. Test-retest reliability was determined using intraclass correlation coefficients (ICC = 0.93, p < 0.01). CONCLUSIONS: The ST is an internally consistent, reliable and valid screening test for cognitive impairment in persons exhibiting early symptoms of dementia. In addition, the ST requires minimal nonprofessional supervision and may be administered by an untrained person. PMID- 16246260 TI - Wnt5 signaling in vertebrate pancreas development. AB - BACKGROUND: Signaling by the Wnt family of secreted glycoproteins through their receptors, the frizzled (Fz) family of seven-pass transmembrane proteins, is critical for numerous cell fate and tissue polarity decisions during development. RESULTS: We report a novel role of Wnt signaling in organogenesis using the formation of the islet during pancreatic development as a model tissue. We used the advantages of the zebrafish to visualize and document this process in living embryos and demonstrated that insulin-positive cells actively migrate to form an islet. We used morpholinos (MOs), sequence-specific translational inhibitors, and time-lapse imaging analysis to show that the Wnt-5 ligand and the Fz-2 receptor are required for proper insulin-cell migration in zebrafish. Histological analyses of islets in Wnt5a(-/-) mouse embryos showed that Wnt5a signaling is also critical for murine pancreatic insulin-cell migration. CONCLUSION: Our results implicate a conserved role of a Wnt5/Fz2 signaling pathway in islet formation during pancreatic development. This study opens the door for further investigation into a role of Wnt signaling in vertebrate organ development and disease. PMID- 16246263 TI - The prognosis of depression in late life versus mid-life: implications for the treatment of older adults. PMID- 16246262 TI - Neuroleptic and benzodiazepine use in long-term care in urban and rural Alberta: characteristics and results of an education intervention to ensure appropriate use. AB - OBJECTIVES: To examine the use of psychotropic drugs in 24 rural and urban long term care (LTC) facilities, and compare the effect of an education intervention for LTC staff and family members on the use of psychotropic drugs in intervention versus control facilities. METHODS: Interrupted time series with a non-equivalent no-treatment control group time series. Data on drug use were collected in 24 Western Canadian LTC facilities (10 urban, 14 rural) for three 2-month time periods before and after the intervention. Pharmacy records were used to collect data on drug, class of drug, dose, administration, and start/stop dates. Chart reviews provided demographics, pro re nata (prn) use, and indications for drug use. Subjects comprised 2443 residents living in the 24 LTC facilities during the 1-year study. An average of 796.33 residents (32.7%) received a psychotropic drug. An education intervention on psychotropic drug use in LTC was offered to intervention physicians, nursing staff, pharmacists and family members. RESULTS: Approximately one-third of residents received a psychotropic drug during the study, often for considerable lengths of time. A minority of psychotropic drug prescriptions had a documented reason for their use, and 69.5% of the reasons would be inappropriate under Omnibus Budget Reconciliation Act (OBRA) legislation. Few psychotropic drug prescriptions were discontinued or reduced during the study. More urban LTC residents received neuroleptics and benzodiazepines than their rural counterparts (26.1% vs. 15.7%, and 18.0% vs. 7.6%, respectively). The education intervention did not result in any significant decline in the use of these drugs in intervention facilities. CONCLUSION: The results suggest substantial use of psychotropic drugs in LTC, although rural LTC residents received approximately half the number of psychotropic drugs compared with urban residents. A resource-intensive intervention did not significantly decrease the use of psychotropics. There is a need for better monitoring of psychotropic drugs in LTC, particularly given that voluntary educational efforts alone may be ineffective agents of change. PMID- 16246264 TI - Effects of high NaCl diet on arterial pressure in Sprague-Dawley rats with hepatic and sinoaortic denervation. AB - The Na(+) receptor that exists in the hepatoportal region plays an important role in postprandial natriuresis and the regulation of Na(+) balance during NaCl load. Thus it would be considered that a dysfunction of the hepatic Na(+) receptor might result in the elevation of arterial pressure under a condition of high NaCl diet. To elucidate this hypothesis, arterial pressure was continuously measured during three weeks of high NaCl diet (8% NaCl) in four groups of rats: (i) intact rats, (ii) rats with hepatic denervation (HD), (iii) rats with sinoaortic denervation (SAD), and (iv) rats with SAD+HD. During a 1-week normal NaCl diet period, there was no difference in arterial pressure among the four groups. A high NaCl diet had no influence on arterial pressure in intact or HD rats; however, it significantly increased by 11 +/- 3 mmHg in SAD rats. The addition of HD to SAD had no synergistic effect on arterial pressure; i.e., in SAD+HD rats, mean arterial pressure increased by 13 +/- 1 mmHg. In conclusion, sinoaortic baroreceptor, but not hepatic Na(+) receptor, has a significant role in the long term regulation of arterial pressure on a high NaCl diet. PMID- 16246265 TI - The role of apoptosis in the development and function of T lymphocytes. AB - Apoptosis plays an essential role in T cell biology. Thymocytes expressing nonfunctional or autoreactive TCRs are eliminated by apoptosis during development. Apoptosis also leads to the deletion of expanded effector T cells during immune responses. The dysregulation of apoptosis in the immune system results in autoimmunity, tumorogenesis and immunodeficiency. Two major pathways lead to apoptosis: the intrinsic cell death pathway controlled by Bcl-2 family members and the extrinsic cell death pathway controlled by death receptor signaling. These two pathways work together to regulate T lymphocyte development and function. PMID- 16246266 TI - Systemic delivery of full-length C/EBP beta/liposome complex suppresses growth of human colon cancer in nude mice. AB - C/EBP beta (CCAAT/enhancer-binding protein beta) is an important transcription factor involved in cellular proliferation and differentiation. Overexpression of the full-length C/EBP beta protein results in cellular growth arrest and apoptosis. Using a nonviral liposome as carrier, we delivered the full-length C/EBP beta expression plasmid, pCN, into nude mice bearing CW-2 human colon cancer tumors via tail vein. Southern blots revealed that the major organs and tumors were transfected. Experimental gene therapy showed that a strong suppression of tumor growth was observed in the pCN-treated mice, and such suppression was due to the overexpression of C/EBP beta, leading to the increased apoptosis in tumors of pCN-treated mice. No apparent toxic effects of pCN/liposome complex were observed in the animals. Thus, C/EBP beta has tumor suppression effect in vivo and may be used in gene therapy for cancers. PMID- 16246267 TI - Regulation of survivin and CDK4 by Epstein-Barr virus encoded latent membrane protein 1 in nasopharyngeal carcinoma cell lines. AB - Latent membrane protein 1 (LMP1), an important protein encoded by Epstein Barr virus (EBV), has been implied to link with the pathogenesis of nasopharyngeal carcinoma (NPC). Its dual effects of increasing cell proliferation and inhibiting cell apoptosis have been confirmed. In this study, we showed that the expression of Survivin and CDK4 protein in CNE-LMP1, a LMP1 positive NPC epithelial cell line, is higher than in LMP1 negative NPC epithelial cell line-CNE1, and the expression is LMP1 dosage-dependent. Although it was reported that Survivin specifically expressed in cell cycle G2/M phase, our studies suggested that LMP1 could promote the expression of Survivin in G0/G1, S and G2/M phase. It also showed that Survivin and CDK4 could be accumulated more in the nuclei triggered by LMP1. More interestingly, Survivin and CDK4 could form a protein complex in the nuclei of CNE-LMP1 rather than in that of CNE1, which demonstrated that the interaction between these two proteins could be promoted by LMP1. These results strongly suggested that the role of LMP1 in the regulation of Survivin and CDK4 may also shed some light on the mechanism research of LMP1 in NPC. PMID- 16246268 TI - Purification of full-length human pregnane and xenobiotic receptor: polyclonal antibody preparation for immunological characterization. AB - Pregnane and Xenobiotic Receptor (PXR; or Steroid and Xenobiotic Receptor, SXR), a new member of the nuclear receptor superfamily, is thought to modulate a network of genes that are involved in xenobiotic metabolism and elimination. To further explore the role of PXR in body's homeostatic mechanisms, we for the first time, report successful prokaryotic expression and purification of full length PXR and preparation of polyclonal antibody against the whole protein. The full-length cDNA encoding a 434 amino acids protein was sub-cloned into prokaryotic expression vector, pET-30b and transformed into E. coli BL21(DE3) cells for efficient over expression. The inclusion body fraction, containing the expressed recombinant protein, was purified first by solubilizing in sarcosine extraction buffer and then by affinity column chromatography using Ni-NTA His Bind matrix. The efficacy of anti-PXR antibody was confirmed by immunocytology, Western blot analysis, EMSA and immunohistochemistry. The antibody obtained was capable of detecting human and mouse PXR with high specificity and sensitivity. Immunofluorescence staining of COS-1 cells transfected with human or mouse PXR showed a clear nuclear localization. Results from immunohistochemistry showed that level of PXR in liver sections is immunologically detectable in the nuclei. Similar to exogenously transfected PXR, Western blot analysis of cell extract from HepG2 and COLO320DM cells revealed a major protein band for endogenous PXR having the expected molecular weight of 50 kDa. Relevance of other immunodetectable bands with reference to PXR isoforms and current testimony are evaluated. Advantages of antibody raised against full-length PXR protein for functional characterization of receptor is discussed and its application for clinical purposes is envisaged. PMID- 16246269 TI - Salt-responsive genes in rice revealed by cDNA microarray analysis. AB - We used cDNA microarrays containing approximately 9,000 unigenes to identify 486 salt responsive expressed sequence tags (ESTs) (representing approximately 450 unigenes) in shoots of the highly salt-tolerant rice variety, Nona Bokra (Oryza sativa L. ssp. Indica pv. Nona). Some of the genes identified in this study had previously been associated with salt stress. However the majority were novel, indicating that there is a great number of genes that are induced by salt exposure. Analysis of the salt stress expression profile data of Nona provided clues regarding some putative cellular and molecular processes that are undertaken by this tolerant rice variety in response to salt stress. Namely, we found that multiple transcription factors were induced during the initial salt response of shoots. Many genes whose encoded proteins are implicated in detoxification, protectant and transport were rapidly induced. Genes supporting photosynthesis were repressed and those supporting carbohydrate metabolism were altered. Commonality among the genes induced by salt exposure with those induced during senescence and biotic stress responses suggests that there are shared signaling pathways among these processes. We further compared the transcriptome changes of the salt-sensitive cultivar, IR28, with that of Nona rice. Many genes that are salt responsive in Nona were found to be differentially regulated in IR28. This study identified a large number of candidate functional genes that appear to be involved in salt tolerance and further examination of these genes may enable the molecular basis of salt tolerance to be elucidated. PMID- 16246270 TI - The presence of ANP in rat peritoneal mast cells. AB - Atrial natriuretic peptide (ANP) is an important component of the natriuretic peptide system. A great role in many regulatory systems is played by mast cells. Meanwhile involvement of these cells in ANP activity is poorly studied. In this work, we have shown the presence of ANP in rat peritoneal mast cells. Pure fraction of mast cells was obtained by separation of rat peritoneal cells on a Percoll density gradient. By Western blotting, two ANP-immunoreactive proteins of molecular masses of 2.5 kDa and 16.9 kDa were detected in lysates from these mast cells. Electron microscope immunogold labeling has revealed the presence of ANP immunoreactive material in storage, secreting and released granules of mast cells. Our findings indicate the rat peritoneal mast cells to contain both ANP prohormone and ANP. These both peptides are located in mast cell secretory granules and released by mechanism of degranulation. It is discussed that many mast cell functions might be due to production of natriuretic peptides by these cells. PMID- 16246273 TI - The weight-loss-at-any-cost environment: how to thrive with a health-centered focus. AB - Although scientists are supposed to be objective and unbiased, and science itself is held as the epitome of objectivity, scientists often force data to fit their preexisting paradigm or deny the existence of data that fall outside their paradigm. The Health At Every Size (HAES) paradigm lies outside the traditional obesity treatment paradigm. However, there are ample data to support the notion that people can be healthy at every size and that weight loss is not necessary for improved health. This article explains how each of us who remains surrounded by the weight-loss-at-any-cost environment can implement HAES practices into our work without additional investment of resources. Techniques are discussed as to how we can incorporate measures other than weight and size into our treatment outcomes, how we can be open-minded to research and clinical practices that are outside our preconceived notions of how things should be, how we can rid ourselves of the assumption that everyone responds to treatment in the same manner, how we can explore clients' feelings to discover the root of their behaviors, and how we can go beyond teaching clients how to behave and teach them how to change behavior. PMID- 16246274 TI - Promoting healthy weight: lessons learned from WIN the Rockies and other key studies. AB - In contrast to the traditional weight-centered approach, the Health At Every Size (HAES) or nondieting approach is health centered, with no focus on losing a predetermined amount of weight or fat. A key HAES principle of advocating healthy changes in food selection rather than adherence to prescriptive diets that involve calorie counting was adopted by Wellness in the Rockies (WIN the Rockies), a community-based research, intervention, and outreach project that promoted healthy lifestyles related to food, physical activity, and body image at the individual and community levels in Wyoming, Montana, and Idaho. The results from the project's cross-sectional surveys indicated that increased frequency of eating food while doing another activity, of drinking sweetened beverages such as soft drinks, and of consuming foods from fast-food restaurants were significant predictors of a high body mass index (BMI). In terms of energy expenditure, other predictors of high BMI from the WIN the Rockies cross-sectional surveys were lower frequency of participation in physical activity and the perception of not getting as much exercise as needed. The overall data provide support for the view that small diet- and physical activity-related lifestyle changes can cumulatively make a significant contribution to maintenance of healthy body weights. Although the community intervention emphasis of WIN the Rockies did not allow a specific assessment of the efficacy of HAES for individual participants in the project, this approach appears to hold great potential for promoting healthful lifestyle changes that improve quality of life. PMID- 16246275 TI - Rethinking weight/health strategies: impact of a convincing guest lecturer. PMID- 16246276 TI - A new you: health for every body: helping adults adopt a health-centered approach to well-being. PMID- 16246277 TI - The meaning of food in our lives: a cross-cultural perspective on eating and well being. AB - Humans are biologically adapted to their ancestral food environment in which foods were dispersed and energy expenditure was required to obtain them. The modern developed world has a surplus of very accessible, inexpensive food. Amid the enormous variety of different foods are "super" foods, such as chocolate, which are particularly appealing and calorie dense. Energy output can be minimal to obtain large amounts of food. In terms of education (eg, in nutrition and risk benefit thinking) and environment design, modern cultures have not kept pace with changes in the food world. Overweight and worrying about food result from this mismatch between human biological predispositions and the current food environment. The French have coped with this mismatch better than Americans. Although at least as healthy as Americans, they focus more on the experience of eating and less on the health effects of eating. They spend more time eating, but they eat less, partly because of smaller portion sizes. French traditions of moderation (versus American abundance), focus on quality (versus quantity), and emphasis on the joys of the moment (rather than making life comfortable and easy) support a healthier lifestyle. The French physical environment encourages slow, moderate social eating, minimal snacking, and more physical activity in daily life. PMID- 16246278 TI - "What's in the Package?" Rethinking grab-and-go lunches. PMID- 16246279 TI - Battling America's epidemic of physical inactivity: building more walkable, livable communities. AB - The US surgeon general recommends a minimum of 30 minutes of physical activity a day for adults to reduce the risk of chronic disease and an early death. Yet only about 1 in 4 American adults meets that recommendation through leisure-time physical activity and/or conscious exercise. One result is the so-called obesity epidemic, but a sedentary lifestyle also increases the risk of chronic diseases, such as diabetes, cardiovascular disease, osteoporosis, and cancer. We must create environments in which physical activity becomes a routine part of the day for more Americans. Encouraging routine walking and bicycling appears to be especially promising because of a growing understanding of how to create bicycle- and pedestrian-friendly settings. Everyone, not just those working in public health, must ally themselves with community leaders to pursue programs and policies to create settings with 4 key attributes: more compact neighborhoods with a mix of land uses; a comprehensive network of pathways, trails, bike lanes, and mass transit to allow "active" transportation; site designs that welcome cyclists and pedestrians; and an umbrella of safety that encourages people to get out of their cars. Many specific resources and programs are recommended to advance this agenda. Finally, we all must become role models by walking and cycling whenever possible and inviting others to do so with us. PMID- 16246280 TI - Walking the talk: incorporating physical activity into conferences and meetings. AB - Have you ever attended a conference that promotes the importance of active living, only to discover that you have spent most of the conference sitting for long periods of time? As was done in the first 2 conferences in 2000 and 2003, Shaping a Healthy Future III: A Rocky Mountain Conference on Weight Realities (2005) incorporated physical activities throughout the meeting to promote active living and to demonstrate how to build enjoyable body movement into the daily lives of busy people. The ideas shared here not only work for conferences but also can be used in staff and committee meetings and even for short breaks at work. PMID- 16246281 TI - An artist's perspective on body image, the media, and contemporary society. AB - The purpose of The Body Image Project is to help in the fight to change the way we look at ourselves and at others. By using the human body as subject matter, I am trying to focus on aesthetic appreciation of the human form and to deal with self-worth by exploring the concept of beauty. Unfortunately, our culture has become obsessed with the image and keeping up appearances. Furthermore, we are letting the so-called beauty industry and corporate America define reality for us, and encouraging body hatred is an extremely lucrative business. But there are more valid and health-promoting ways to define beauty and reality. We must educate ourselves to recognize that advertising is not a slice of reality. We need to quit giving other people permission to define who we are and what we are worth. We need to begin to see healthy bodies as the ideal, and we need to make clear that good health is not defined by size; it is a state of physical, mental, and social well-being. Change can happen, but it will take a concerted effort by many people. This project needs your help. PMID- 16246282 TI - Preventing the broad spectrum of weight-related problems: working with parents to help teens achieve a healthy weight and a positive body image. AB - A spectrum of eating-, activity-, and weight-related concerns is presented that includes 5 dimensions (weight control practices, level of physical activity, body image, eating behaviors, and weight status) and different levels of severity within each of these dimensions. Multiple interacting factors contribute to the etiology of problems within each of these dimensions in adolescents at the individual, familial, peer, school, community, and societal levels. Families have an important role to play in reinforcing the positive influences at each of these levels and in filtering out the negative influences. Parents can help their children engage in more healthful eating and physical activity behavior and feel better about themselves through (1) role modeling healthful behaviors, (2) providing an environment that makes it easy for their children to make healthful choices, (3) focusing less on weight and more on behaviors and overall health, and (4) providing a supportive environment for their children to enhance communication. Families need to be proactive within our society, which works against the development of a healthy weight and a positive body image in children and adolescents. However, families cannot do it on their own and need support from the more distal environments within which they function. PMID- 16246283 TI - Full of ourselves: a wellness program to advance girl power, health and leadership: an eating disorders prevention program that works. PMID- 16246284 TI - Wonders within WIN Wyoming. PMID- 16246285 TI - Shaping a Healthy Future III: A Rocky Mountain Conference on Weight Realities Jackson, Wyoming, USA, April 27-29, 2005. Proceedings and abstracts. PMID- 16246286 TI - A method for identifying rear-end collision risks using inductive loop detectors. AB - An innovative feature of this study is to firstly attempt to capture rear-end collision potentials from the analysis of inductive loop detector data. Signals collected from loops are applied for monitoring individual vehicle information on freeways to estimate safe stopping distances in car-following situations. An index to quantify the potential of rear-end collisions is derived, and further employed for developing criteria to evaluate levels of rear-end collision risks. The proposed methodology based on loop detector data enables to identify collision potentials in real time. It is believed that the index would be a valuable tool for operating agencies in developing various strategies and policies toward enhancements of traffic safety. PMID- 16246287 TI - A critical examination of arguments against bicycle helmet use and legislation. AB - In a recent paper published in Accident Analysis & Prevention, Curnow puts forward a number of arguments against legislating bike helmet use [Curnow, W.J., 2005 The Cochrane Collaboration and bicycle helmets. Accid. Anal. Prevent. 37(3), 569-573]. He begins by criticizing the scientific evidence that helmets protect against head and brain injuries. The crux of his argument is that in theory helmets should not protect all mechanisms of brain injury and, therefore, all epidemiological research showing they are beneficial in a variety of circumstances is invalid. This short communication identifies some of the questionable elements in Curnow's assertions. PMID- 16246288 TI - Isolation, purification and characterization of GPI-anchored membrane proteins from Trypanosoma rangeli and Trypanosoma cruzi. AB - GPI-anchored proteins from plasma membrane of Trypanosoma rangeli and Trypanosoma cruzi epimastigotes were isolated and characterized using the partition Triton X 114 method. The detection by Western blot of specific proteins of 90, 85 and 56 kDa molecular mass in T. rangeli compared to those of 30, 70 and 100 kDa detected in T. cruzi demonstrates specific discrimination between these two species of Trypanosoma. The potential diagnostic value of the here reported proteins to differentiate mixed infections by T. cruzi and T. rangeli is evaluated and its potential for epidemiological studies of Chagas disease in endemic areas is also discussed. PMID- 16246289 TI - Stemless self-quenching reporter molecules identify target sequences in mRNA. AB - The design of oligonucleotides for gene silencing requires a rational method for identifying hybridization-accessible sequences within the target RNA. To this end, we have developed stem-loop self-quenching reporter molecules (SQRMs) as probes for such sequence. SQRMs have a 5' fluorophore, a quenching moiety on the 3' end, an intervening sequence that forms an approximately 5-basepaired stem, and a loop sequence of approximately 20-30 bases. We have previously described a mapping strategy employing SQRMs to locate stem-loop structures in the target mRNA molecule. We now show that the original design constraint of a basepaired stem is not needed, either in vitro or in vivo. We propose that stemless probes possess sufficient signal-to-noise for use in vivo and that this ratio is an indication of hybridization of the probe to its target. Data showing that these SQRMs can specifically target and reduce c-Myb protein synthesis and can be used for real-time in vivo assays are presented. PMID- 16246290 TI - Synthesis and study of the electrophoretic behavior of mannan conjugates with cyclic peptide analogue of myelin basic protein using lysine-glycine linker. AB - New approaches for the treatment of multiple sclerosis involve the design and synthesis of peptide or nonpeptide analogues of myelin sheath, which could alter the immune response of patients. For this purpose, the cyclo(75-82) myelin basic protein (MBP)(74-85) analogue was conjugated to mannan (a polymannose) via (Lys Gly)(5) linker. Monitoring of synthesis of the (Lys-Gly)(5)-containing cyclic analogue of MBP, mannan oxidation, and the conjugation reaction of this analogue to oxidized mannan was performed with capillary electrophoresis (CE) in operating buffers of different pH values. The (Lys-Gly)(5)cyclo(75-82)MBP(74-85) was efficiently synthesized by solid-phase synthesis and purified to a high degree, as confirmed by CE analysis in a low-pH (3.0) phosphate buffer and normal polarity. Oxidation of mannan was monitored using a high-pH (9.3) borate buffer, and the generation of heterogeneous products and even UV-absorbing peaks was shown by CE. CE analysis in a pH 5.1 phosphate buffer offers high resolution of oxidized mannan and the conjugation product and can be used for screening of the reaction products. Mannan-peptide conjugates of varying degrees of substitution and unreacted mannan were observed. The developed CE analysis presents distinct advantages over sodium dodecyl sulfate-polyacrylamide gel electrophoresis such as high versatility, high separation efficiency, short analysis time, low cost, and low solvent consumption. PMID- 16246291 TI - An ultrasensitive fluorescent assay for the in vivo quantification of superoxide radical in organisms. AB - Superoxide radical is a very important parameter of oxidative stress involved in a variety of biological phenomena; therefore, its in vivo study is of utmost significance. However, its accurate detection is a challenge due to its short lifetime and its very low physiological concentration. All current assays are qualitative and nonspecific, and at best they are performed in vitro. The current dihydroethidine-based assay overcomes all these problems and introduces the following novelties. First, it measures the in vivo superoxide production in animals, plants, and microorganisms. Second, it is ultrasensitive and very simple in that it can measure superoxide radical as low as 1.5 pmol in biological samples as low as 5 mg. Third, the very high sensitivity of the assay renders possible, for the first time, the measurement of the actual rate of formation of superoxide radical under physiological and simulated nonphysiological conditions. PMID- 16246292 TI - A live cell hormone-binding assay on transgenic bacteria expressing a eukaryotic receptor protein. AB - The investigation of hormone-receptor interaction normally needs isolation and extensive purification of the receptor protein or a particular receptor containing fraction. To bypass these time- and resource-consuming procedures, we have established a live cell-based assay using transgenic bacteria expressing single eukaryotic receptors. Here we describe some biochemical features of the Arabidopsis cytokinin receptor CRE1/AHK4 expressed in Escherichia coli. The data show that the main characteristics of the ligand-receptor interaction, including binding affinity and ligand specificity, can be determined using intact bacteria expressing a functional receptor. PMID- 16246293 TI - [Preoperative location of microcalcifications after macrobiopsy: failure of lipiodol]. AB - Accurate mark of macrobiopsy site, carried out borderline or malignant lesions, is very important for surgeons. We report a woman case, 68 year's old, who presents intraductal carcinoma diagnoses on macrobiopsy. On postbiopsy X-ray, we can note lipiodol used instead of clip to reaper macrobiopsy site. Our observation shows how lipiodol use is not adapted into this indication. PMID- 16246294 TI - [Hepatic haemangioendothelioma: case report and review of literature]. AB - A 37 year old-woman complained about persistant pain of right abdominal quadrant. We discovered many hepatic lesions. Histology revealed benign processus. Evolution was marked by peritoneal carcinomatosis. After reexamination of histology we discovered hepatic haemangioendothelioma. The patient died 18 month after diagnosis although chemotherapy. PMID- 16246295 TI - [Primary hepatic lymphoma of MALT-type: a tumor that can simulate a liver metastasis]. AB - Primary hepatic lymphomas are rare tumors. We report a case of a 72 year-old woman with a past history of colonic adenocarcinoma who presented primary hepatic lymphoma of MALT-type. The patient had been operated on 3 years before for colonic adenocarcinoma, pT3N0, revealed by a bowel obstructive syndrome. She had been treated by chemotherapy for 6 months. During the follow-up, the computed tomography-scan (CT-scan) revealed the presence of a not well-demarcated mass in segment III of the liver, measuring 4 cm in diameter. The tumor was hypodense and was not enhanced on dynamic study. The mass was already present on the initial CT scan. Left lobectomy was performed with the diagnosis of liver metastasis of the colonic adenocarcinoma. Surgical specimen showed a tumor composed of a dense infiltrate of small lymphocytes positive for B-cell markers on immunohistochemistry. The tumor contained reactive lymphoid follicles and there were numerous lympho-epithelial biliary lesions. The patient is alive and free of disease 2 years after the diagnosis. Primary hepatic lymphoma of MALT-type is a low-grade B cell lymphoma. Twenty-five cases had been reported in the literature so far. The patients were 16 females and 9 males, mean age 63.5 years. The pathogenesis is still unclear but half of the patients had a past history of chronic inflammatory liver disease (hepatitis B or C virus infection, ascaris infection, primary biliary cirrhosis) or malignant neoplasm. This tumor has a good prognosis; it is usually limited to the liver and surgical resection cures the patient in most cases. PMID- 16246296 TI - Simulated solar irradiation with enhanced UV-B adjust plastid- and thylakoid associated polyamine changes for UV-B protection. AB - Polyamines have been described to protect against numerous oxidative stresses in plants. Increasing UV-B radiation (280-315 nm) in the biosphere may also induce an increase in radical formation in tissues. This study employed the tobacco cultivars Bel B and Bel W3 to describe possible protective functions of polyamines against UV-B radiation in sun light simulators (GSF/Munich) with natural diurnal fluctuations of simulated UV-B. Polyamine measurements on a whole leaf basis in isolated chloroplasts and thylakoids were paralleled to photosynthetic and respiration rates, photosynthetic efficiency, leaf thickness and photosynthetic pigment compositions. The study revealed that an increase of polyamines, and especially of putrescine level in thylakoid membranes upon elevated UV-B exposure comprises one of the primary protective mechanisms in the photosynthetic apparatus of the tobacco variety Bel B against UV-B radiation. The tobacco cultivar Bel W3, sensitive to ozone, was also proved to be sensitive to UV-B. This sensitivity is attributed to its incapability to enhance putrescine level in thylakoid membranes. After prolongation of UV-B exposure, when endogenous plant balances are being gradually restored, due to secondary responses, (e.g., biosynthesis of carotenoids and of additional flavonoids) and the plant is adapting to the altered environmental conditions, then the polyamine level is being reduced. Thus, we can discriminate the UV-B induced stress period from a UV-B acclimation period. PMID- 16246297 TI - Type 1 ryanodine receptor in cardiac mitochondria: transducer of excitation metabolism coupling. AB - Mitochondria in a variety of cell types respond to physiological Ca(2+) oscillations in the cytosol dynamically with Ca(2+) uptakes. In heart cells, mitochondrial Ca(2+) uptakes occur by a ruthenium red-sensitive Ca(2+) uniporter (CaUP), a rapid mode of Ca(2+) uptake (RaM) and a ryanodine receptor (RyR) localized in the inner mitochondrial membrane (IMM). Three subtypes of RyRs have been described and cloned, however, the subtype identity of the mitochondrial ryanodine receptor (mRyR) is unknown. Using subtype specific antibodies, we characterized the mRyR in the IMM from rat heart as RyR1. These results are substantiated by the absence of RyR protein in heart mitochondria from RyR1 knockout mice. The bell-shape Ca(2+)-dependent [(3)H]ryanodine binding curve and its modulation by caffeine and adenylylmethylenediphosphonate (AMPPCP) give further evidence that mRyR functions pharmacologically like RyR1. Ryanodine prevents mitochondrial Ca(2+) uptake induced by raising extramitochondrial Ca(2+) to 10 microM. Similarly, ryanodine inhibits oxidative phosphorylation stimulated by 10 microM extramitochondrial Ca(2+). In summary, our results show that the mRyR in cardiac muscle has similar biochemical and pharmacological properties to the RyR1 in the sarcoplasmic reticulum (SR) of skeletal muscle. These results could also suggest an efficient mechanism by which mitochondria sequesters Ca(2+) via mRyR during excitation-contraction coupling to stimulate oxidative phosphorylation for ATP production to meet metabolic demands. Thus, the mRyR functions as a transducer for excitation-metabolism coupling. PMID- 16246298 TI - Inhibition of crotoxin binding to synaptosomes by a receptor-like protein from Crotalus durissus terrificus (the South American rattlesnake). AB - Crotoxin (Ctx) is a potent neurotoxin of the venom of Crotalus durissus terrificus (the South American rattlesnake). Ctx is a heterodimer composed of CB, a toxic PLA(2) subunit, and CA, a non-toxic and non-enzymatic subunit, that potentiates the neurotoxicity of CB in vivo. The deleterious action of Ctx upon C. d. terrificus snakes themselves is known to be prevented by a PLA(2) inhibitor (CNF) present in their blood serum. CNF acts by replacing CA in Ctx, thus forming a new stable complex CNF-CB. This complex no longer interacts with the target receptor (TR) to deliver CB to cause its lethal effect. Furthermore, CNF-CB seems to be reminiscent of the interaction Ctx-TR at the pre-synaptic site. In the present work, the binding competition between rat brain synaptosomes (TR) and CNF for Ctx was investigated. Radiolabeled Ctx, made of CA and one isoform of CB (CA (125)ICB(2)), was used as ligand. The competition by unlabeled Ctx was taken as a reference. The potency of CNF as a competitor was evaluated under different incubation conditions with varying time scale addition of reagents (CA (125)ICB(2), synaptosomes and CA-CB(2) or CNF). CNF was able to inhibit the binding of the toxin to synaptosomes as well as to partially displace the toxin already bound to its membrane target. The mechanisms of competition involved were discussed and a previous schematic model of interactions between Ctx, TR and CNF was updated. PMID- 16246299 TI - Biotinylation of heat shock protein 70 induces RANTES production in HEK293 cells in a CD40-independent pathway. AB - Biotinylated proteins and peptides have been used as popular ligands for characterization of cell surface receptors by a variety of methods including flow cytometry. The number and the location of biotin moieties incorporated could alter the structural and physicochemical properties of ligands, although biotin is thought to be such a small molecule (244Da) that it is capable of being conjugated to most proteins without affecting their activity. Here, we demonstrate that the biotinylated HSP70 molecule via primary amines bound to epithelium-like HEK 293 cells in a saturable manner whereas the unlabeled counterparts of HSP70 other than mouse Hsp72 do not. This binding was not competed by either HSP70 or the biotin entity itself. Interestingly, the biotinylated HSP70 also elicited the production of CC-chemokine RANTES independent of CD40 signaling. This response occurred regardless of sequence diversity of HSP70 derived from different species, and neither the biotinylated ovalbumin nor the unlabeled HSP70 cross-linked with a biotinylated protein stimulated a significant level of RANTES production which was induced by biotinylated HSP70 itself. Our findings suggest that modification of HSP70 such as biotinylation may function as a biological alarm signal in the innate immune system. PMID- 16246300 TI - The reversibility of the glutathionyl-quercetin adduct spreads oxidized quercetin induced toxicity. AB - Quercetin is one of the most prominent dietary antioxidants. During its antioxidant activity, quercetin becomes oxidized into its o-quinone/quinone methide QQ. QQ is toxic since it instantaneously reacts with thiols of, e.g., proteins. In cells, QQ will initially form an adduct with glutathione (GSH), giving GSQ. We have found that GSQ is not stable; it dissociates continuously into GSH and QQ with a half life of 2min. Surprisingly, GSQ incubated with 2 mercapto-ethanol (MSH), a far less reactive thiol, results in the conversion of GSQ into the MSH-adduct MSQ. A similar conversion of GSQ into relatively stable protein thiol-quercetin adducts is expected. With the dithiol dihydrolipoic acid (L(SH)(2)), quercetin is formed out of GSQ. These results indicate that GSQ acts as transport and storage of QQ. In that way, the initially highly focussed toxicity of QQ is dispersed by the formation of GSQ that finally spreads QQ induced toxicity, probably even over cells. PMID- 16246301 TI - Effect of W62G mutation of hen lysozyme on the folding in vivo. AB - In previous paper, we showed that W62G mutation caused ill effects at the early stages of folding of the reduced hen lysozyme in vitro. Here, we investigated whether the single mutation brings about drastic turn to in vivo folding of lysozyme. W62G lysozyme was secreted from yeast cells and then purified with ion exchange chromatography. From the results of gel chromatography and peptide analysis, the species with two cysteines, Cys80 and Cys94, and non-native cystine, Cys64-Cys76, was partially present in secreted product of yeast containing gene for W62G lysozyme. Thus, it was suggested that W62G mutation also affected the in vivo folding of lysozyme. PMID- 16246302 TI - Selective upregulation of immune regulatory and effector cytokine synthesis by intestinal intraepithelial lymphocytes following CD43 costimulation. AB - The involvement of the CD43 molecule in the activation of mouse small intestinal intraepithelial lymphocytes (IELs) has been studied using a panel of twenty-two regulatory and effector immune response analytes. In the absence of stimulation in vitro, IELs produced low levels of CCL5 only. Upon CD3 stimulation, the activity of seven of twenty-two analytes was elevated relative to unstimulated cultures, including several proinflammatory cytokines and chemokines. Notably, CD3 stimulation in the presence of CD43 costimulation resulted in elevated levels of five analytes (interleukin-2, interferon-gamma, CCL5, granulocyte colony stimulating factor, and granulocyte-monocyte colony-stimulating factor) above that produced by CD3 stimulation alone. That CD43 costimulation was responsible for elevated cytokine/chemokine activity was confirmed at the transcriptional level by real-time PCR for IFN-gamma and CCL5, and by ELISA for IFN-gamma. These findings open the way to a better understanding of the process by which T cells are activated in the intestinal epithelium. PMID- 16246303 TI - Bilirubin derived from heme degradation suppresses MHC class II expression in endothelial cells. AB - The enzymatic action of heme oxygenase (HO) is mediated by the cleavage of heme into carbon monoxide, ferrous iron, and biliverdin/bilirubin. Here, we show that induction of HO-1 expression, an inducible form of HO, down-regulates IFN-gamma induced MHC class II expression in endothelial cells. Among three catalytic products of HO, bilirubin, but not carbon monoxide or ferrous iron, mediated the suppressive effects of HO through the reduction of mRNA levels of Stat-1 dependent class II transactivator. Expression of HO-1 could suppress the levels of IFN-gamma-induced Stat-1 phosphorylation. This effect could be mimicked by exposing the cells to one of its catalytic products, bilirubin. In addition, HO-1 or bilirubin could modulate the transcript activities of Stat-1-driven gene expression in luciferase reporter assays. These findings suggest an important role of HO-1 in the modulation of immune responses through suppression of MHC-II expression in antigen presenting cells. Our data provide a new line of evidence supporting HO-1-targeted therapy for immune modulation. PMID- 16246304 TI - Effect of induced expression of an antimicrobial peptide melittin on Chlamydia trachomatis and Mycoplasma hominis infections in vivo. AB - A plasmid construct was designed in which the gene of antimicrobial peptide melittin is controlled by the tetracycline-responsive promoter of human cytomegalovirus, aided by a constitutively expressed trans-activator protein gene. Its vaginal administration and induction of melittin gene transcription with doxycycline markedly suppressed subsequent genital tract infection of mice by Mycoplasma hominis and Chlamydia trachomatis. At least half of the melittin protected animals proved free of either pathogen within 3-4 weeks. Recombinant plasmids expressing genes of antimicrobial peptides hold much promise as agents for prevention and control of urogenital latent infections. PMID- 16246305 TI - Blockade of gamma-secretase activity within the hippocampus enhances long-term memory. AB - The gamma-secretase complex, a membrane-bound aspartyl protease, hydrolyzes the transmembrane domains of several integral membrane proteins including the key signaling molecules amyloid precursor protein (APP), Notch, deleted in colorectal cancer (DCC), and N- and E-cadherins. The proteolysis processing of these proteins is critical for generation of signaling molecules that may participate in neuronal communication and plasticity. Using a potent gamma-secretase inhibitor, L-685,458, we examined if blockade of its activity in the hippocampus can influence contextual and spatial memory in rats. Surprisingly, we observed that post-training blockade of gamma-secretase activity leads to enhanced long term memory in two hippocampus-dependent tasks. This suggests that a signaling molecule(s) generated by gamma-secretase activity may have a negative influence on long-term memory formation. PMID- 16246306 TI - cAMP-response element-binding protein mediates prostaglandin F2alpha-induced hypertrophy of vascular smooth muscle cells. AB - Prostaglandin F(2alpha) (PGF(2alpha)) is a vasoactive factor that causes constriction and hypertrophy of vascular smooth muscle cells (VSMCs). However, the mechanism of PGF(2alpha)-induced hypertrophy is largely unknown. Cyclic AMP response element (CRE)-binding protein (CREB), the best characterized stimulus induced transcription factor, activates transcription of target genes with CRE and promotes cell growth. We examined the role of CREB in PGF(2alpha)-induced hypertrophy of VSMCs. PGF(2alpha) induced phosphorylation of CREB at serine 133, which is a critical marker of activation, after 5-10min of stimulation in a dose dependent manner. Pharmacological inhibition of extracellular signal-regulated protein kinase and p38 mitogen-activated protein kinase (p38-MAPK) suppressed PGF(2alpha)-induced CREB phosphorylation. Inhibition of epidermal growth factor receptor (EGFR) and mitogen- and stress-activated protein kinase-1 also suppressed PGF(2alpha)-induced CREB phosphorylation. Overexpression of dominant negative form of CREB (AdCREB M1), of which serine 133 was replaced with alanine, inhibited PGF(2alpha)-induced c-fos mRNA expression as well as hypertrophy of VSMCs [hypertrophy index (microg/10(4)cell); control 8.13, PGF(2alpha) 9.85, AdCREB M1 7.91, and AdCREB M1+PGF(2alpha) 8.43]. These results suggest that PGF(2alpha) activated CRE-dependent gene transcription through EGFR transactivation, and the CREB pathway plays a critical role in PGF(2alpha) induced hypertrophy of VSMCs. PMID- 16246307 TI - Mechanism of osteogenic induction by FK506 via BMP/Smad pathways. AB - FK506 is an immunosuppressant that exerts effects by binding to FK506-binding protein 12 (FKBP12). Recently, FK506 has also been reported to promote osteogenic differentiation when administered locally or in vitro in combination with bone morphogenetic proteins (BMPs), although the underlying mechanism remains unclarified. The present study initially showed that FK506 alone at a higher concentration (1muM) induced osteogenic differentiation of mesenchymal cell lines, which was suppressed by adenoviral introduction of Smad6. FK506 rapidly activates the BMP-dependent Smads in the absence of BMPs, and the activation was blocked by Smad6. Overexpression of FKBP12, which was reported to block the ligand-independent activation of BMP type I receptor A (BMPRIA), suppressed Smad signaling induced by FK506, but not that induced by BMP2. BMPRIA and FKBP12 bound to each other, and this binding was suppressed by FK506. These data suggest that FK506 promotes osteogenic differentiation by activating BMP receptors through interacting with FKBP12. PMID- 16246308 TI - In vivo interaction between RGS4 and calmodulin visualized with FRET techniques: possible involvement of lipid raft. AB - Regulators of G-protein signaling (RGS) are a family of proteins which accelerate intrinsic GTP-hydrolysis on heterotrimeric G-protein-alpha-subunits. Although it has been suggested that the function of RGS4 is reciprocally regulated by competitive binding of the membrane phospholipid, phosphatidylinositol-3,4,5, trisphosphate(PtdIns(3,4,5)P(3)), and Ca(2+)/calmodulin (CaM), it remains to be shown that these interactions occur in vivo. Here, using fluorescence resonance energy transfer (FRET) techniques, we show that an elevation of intracellular Ca(2+) concentration by ionomycin increased the FRET efficiency from ECFP (a variant of cyan fluorescent protein)-labeled calmodulin to Venus (a variant of yellow fluorescent protein)-labeled RGS4. The increase in FRET efficiency was greatly attenuated by pre-treating the cells with methyl-beta-cyclodextrin, which depletes membrane cholesterol and thus disrupts lipid rafts. These results provide the first demonstration of a Ca(2+)-dependent interaction between RGS4 and CaM in vivo and show that association in lipid rafts of the plasma membrane might be involved in this physiological regulation of RGS proteins. PMID- 16246310 TI - FAD oxidizes the ERO1-PDI electron transfer chain: the role of membrane integrity. AB - The molecular steps of the electron transfer in the endoplasmic reticulum from the secreted proteins during their oxidation are relatively unknown. We present here that flavine adenine dinucleotide (FAD) is a powerful oxidizer of the oxidoreductase system, Ero1 and PDI, besides the proteins of rat liver microsomes and HepG2 hepatoma cells. Inhibition of FAD transport hindered the action of FAD. Microsomal membrane integrity was mandatory for all FAD-related oxidation steps downstream of Ero1. The PDI inhibitor bacitracin could inhibit FAD-mediated oxidation of microsomal proteins and PDI, but did not hinder the FAD-driven oxidation of Ero1. Our data demonstrated that Ero1 can utilize FAD as an electron acceptor and that FAD-driven protein oxidation goes through the Ero1-PDI pathway and requires the integrity of the endoplasmic reticulum membrane. Our findings prompt further studies to elucidate the membrane-dependent steps of PDI oxidation and the role of FAD in redox folding. PMID- 16246309 TI - CPT1alpha over-expression increases long-chain fatty acid oxidation and reduces cell viability with incremental palmitic acid concentration in 293T cells. AB - To test the cellular response to an increased fatty acid oxidation, we generated a vector for an inducible expression of the rate-limiting enzyme carnitine palmitoyl-transferase 1alpha (CPT1alpha). Human embryonic 293T kidney cells were transiently transfected and expression of the CPT1alpha transgene in the tet-on vector was activated with doxycycline. Fatty acid oxidation was measured by determining the conversion of supplemented, synthetic cis-10-heptadecenoic acid (C17:1n-7) to C15:ln-7. CPT1alpha over-expression increased mitochondrial long chain fatty acid oxidation about 6-fold. Addition of palmitic acid (PA) decreased viability of CPT1alpha over-expressing cells in a concentration-dependent manner. Both, PA and CPT1alpha over-expression increased cell death. Interestingly, PA reduced total cell number only in cells over-expressing CPT1alpha, suggesting an effect on cell proliferation that requires PA translocation across the mitochondrial inner membrane. This inducible expression system should be well suited to study the roles of CPT1 and fatty acid oxidation in lipotoxicity and metabolism in vivo. PMID- 16246311 TI - The journey from NADPH-cytochrome P450 oxidoreductase to nitric oxide synthases. AB - This mini-review will reflect the perspective of its author on two fields of research, which have merged as the result of the insights of investigators whose work has influenced both areas immeasurably. It cannot be overlooked, however, that the research activities of many during a period of over five decades have produced the chemical and biological bases for the exciting discoveries now encompassing the cytochromes P450 and their redox partners, and the three isoforms of nitric oxide synthase as they function in their respective biological milieux. Following the remarkable discovery that, indeed, molecular oxygen can be adducted to organic molecules by enzymatic systems and that such processes require a supply of reducing equivalents, it is the purpose of this review to provide a chart, with some of its detours, of the road that followed in the pursuit of interesting biological phenomena involving these two major oxygenation systems. It is not intended to be a balanced review and apologies must be offered in advance to those whose contributions may be overlooked or simply were not directly germane to the development of the author's journey. PMID- 16246312 TI - Epitope shared by functional variant of organic cation/carnitine transporter, OCTN1, Campylobacter jejuni and Mycobacterium paratuberculosis may underlie susceptibility to Crohn's disease at 5q31. AB - Campylobacter jejuni and Mycobacterium paratuberculosis have been implicated in the pathogenesis of Crohn's disease. The presence of bacterial metabolites in the colonic lumen causing a specific breakdown of fatty acid oxidation in colonic epithelial cells has been suggested as an initiating event in inflammatory bowel disease (IBD). l-Carnitine is a small highly polar zwitterion that plays an essential role in fatty acid oxidation and ATP generation in intestinal bioenergetic metabolism. The organic cation/carnitine transporters, OCTN1 and OCTN2, function primarily in the transport of l-carnitine and elimination of cationic drugs in the intestine. High-resolution linkage disequilibrium mapping has identified a region of about 250kb in size at 5q31 (IBD5) encompassing the OCTN1 and -2 genes, to confer susceptibility to Crohn's disease. Recently, two variants in the OCTN1 and OCTN2 genes have been shown to form a haplotype which is associated with susceptibility to Crohn's. We show that OCTN1 and OCTN2 are strongly expressed in target areas for IBD such as ileum and colon. Further, we have now identified a nine amino acid epitope shared by this functional variant of OCTN1 (Leu503Phe) (which decreases the efficiency of carnitine transport), and by C. jejuni (9 aa) and M. paratuberculosis (6 aa). The prevalence of this variant of OCTN1 (Phe503:Leu503) is 3-fold lower in unaffected individuals of Jewish origin (1:3.44) compared to unaffected individuals of non-Jewish origin (1:1). We hypothesize that a specific antibody raised to this epitope during C. jejuni or M. paratuberculosis enterocolitis would cross-react with the intestinal epithelial cell functional variant of OCTN1, an already less efficient carnitine transporter, leading to an impairment of mitochondrial beta-oxidation which may then serve as an initiating event in IBD. This impairment of l-carnitine transport by OCTN1 may respond to high-dose l-carnitine therapy. PMID- 16246313 TI - Ethanol modulates GABA(B) receptor expression in cortex and hippocampus of the adult rat brain. AB - Using in situ hybridization, RNase protection assay and Western blot, we studied the effects of ethanol on the expression levels of GABA(B) receptor mRNA and protein in the cortex and hippocampus from adult rat brain. The results showed that ethanol significantly increased GABA(B1) and GABA(B2) receptor protein expression in the cortex, whereas only GABA(B2) was increased in the hippocampus. GABA(B) receptor agonist baclofen could partially reverse the effect of ethanol. Further studies of the mRNA levels defined that GABA(B1) mRNA levels were significantly increased in the hippocampus, with no significant changes of GABA(B2) mRNA levels. Moreover, GABA(B1) and GABA(B2) receptor mRNA levels were increased on 3-week ethanol treatment. Finally, GABA(B) agonist baclofen and antagonist phaclofen showed significant decreasing effects on GABA(B1) receptor mRNA levels in the cortex, but not in the hippocampus. These results were further confirmed by in situ hybridization. Thus, the present results showed the effects of ethanol on GABA(B) receptors in the cortex and hippocampus, implying the possible role of GABA(B) receptor in ethanol effects. The effects of GABA(B) receptor agonist and antagonist suggested that the possible mechanisms underlying that GABA(B) receptor modulated the behavioral effect induced by ethanol. PMID- 16246314 TI - Adenosine A1 receptors mediate inhibition of cAMP formation in vitro in the pontine, REM sleep induction zone. AB - Microinjection of adenosine A1 receptor agonist or an inhibitor of adenylyl cyclase into the caudal, oral pontine reticular formation (PnOc) of the rat induces a long-lasting increase in REM sleep. Here, we report significant inhibition of forskolin-stimulated cAMP in dissected pontine tissue slices containing the PnOc incubated with the A1 receptor agonist, cyclohexaladenosine (10(-8) M). These data are consistent with adenosine A1 receptor agonist actions on REM sleep mediated through inhibition of cAMP. PMID- 16246316 TI - Correlation between bioprotective effectiveness and dynamic properties of trehalose-water, maltose-water and sucrose-water mixtures. AB - The aim of the present work is to link the bioprotective effectiveness to the dynamic properties of a class of homologous disaccharides, that is, trehalose, maltose and sucrose, and their mixtures in water. The findings obtained by elastic neutron scattering point out a harmonic-anharmonic transition for all the three disaccharide mixtures. Using a new operative definition of 'fragility', the different degrees of 'strength' of the investigated systems are determined. The links existing between the degree of fragility and the cryptoprotective action are also discussed. PMID- 16246317 TI - Mapping enzymatic functionalities of mannuronan C-5 epimerases and their modular units by dynamic force spectroscopy. AB - Alginates are (1-->4)-linked structural copolyuronans consisting of beta-D mannuronic acid (M) and its C-5 epimer alpha-L-guluronic acid (G). The residue sequence variation is introduced in a unique postpolymerisation step catalysed by a family of C-5 epimerases named AlgE enzymes. The seven known AlgE's are composed of two modules, designated A and R, present in different number. The molecular details of the structure-function relationship of these seven epimerases, introducing specific residue sequences, are not understood. In this study, single-molecular pair interactions between alginate and AlgE enzymes were investigated using dynamic force spectroscopy. The AlgE enzymes AlgE4 and AlgE6, the recombinant construct PKA1 composed of A- and R-modules from various AlgE's, as well as separate R- and A-modules were studied. The strength of the protein mannuronan interaction, when applying a loading rate of 0.6 nN/s, varied from 73 pN (AlgE4) to 144 pN (A-module). The determined potential width, that is, the distance from the activation barrier to the bound substrate molecule, was 0.23 nm for AlgE4, 0.19 nm for AlgE6 and 0.1 nm for the A-module. No attraction was observed between the R-module and the substrate. The observations indicate that the A-module contains the substrate binding site and that the R-module modulates the enzyme-substrate binding strength. The observed AlgE4-polymer residence times, two orders of magnitude longer than expected from kcat reported for AlgE4, not observed for PKA1, led us to propose a processive mode of action of AlgE4. PMID- 16246318 TI - On the multifunctionality of cholinesterases. AB - Here we present novel information on non-classical functions of cholinesterases and on a cross-talk linking the two enzymes AChE and BChE. The first part of the article is focussed on the regulation of ChEs and the effects acquired when one of the proteins is knocked down (siRNA for BChE, AChE knock-out mouse). In the second part evidence is presented showing that AChE may exert adhesive properties through its binding to laminin, thus being involved in cell-matrix or cell-cell communication. PMID- 16246319 TI - Infants' categorization of novel objects with more or less obvious features. AB - In five experiments, 14- to 15-month-old infants' categorization of objects on the basis of more or less obvious features was investigated. Using an object examining paradigm, a total of 200 infants were familiarized with novel objects that shared either more obvious features (i.e., easily visible) or less obvious features (i.e., accessible by lifting a flap), followed by an in-category object and an out-of-category object. When only perceptual information was available, infants formed a category on the basis of the more obvious features but not on the basis of the less obvious features (Experiments 1 and 3). When infants were provided with animacy cues and/or object names, they formed categories on the basis of either more or less obvious features (Experiments 2, 4, and 5). The results of these studies delineate the role of animacy cues and object names in establishing categories on the basis of less obvious features. PMID- 16246320 TI - Unsupervised clustering in mRNA expression profiles. AB - The development of microarray technologies gives scientists the ability to examine, discover and monitor the mRNA transcript levels of thousands of genes in a single experiment. Nonetheless, the tremendous amount of data that can be obtained from microarray studies presents a challenge for data analysis. The most commonly used computational approach for analyzing microarray data is cluster analysis, since the number of genes is usually very high compared to the number of samples. In this paper, we investigate the application of the recently proposed k-windows clustering algorithm on gene expression microarray data. This algorithm apart from identifying the clusters present in a data set also calculates their number and thus requires no special knowledge about the data. To improve the quality of the clustering, we employ various dimension reduction techniques and propose a hybrid one. The results obtained by the application of the algorithm exhibit high classification success. PMID- 16246321 TI - Of chicken wings and frog legs: a smorgasbord of evolutionary variation in mechanisms of tetrapod limb development. AB - The tetrapod limb, which has served as a paradigm for the study of development and morphological evolution, is becoming a paradigm for developmental evolution as well. In its origin and diversification, the tetrapod limb has undergone a great deal of remodeling. These morphological changes and other evolutionary phenomena have produced variation in mechanisms of tetrapod limb development. Here, we review that variation in the four major clades of limbed tetrapods. Comparisons in a phylogenetic context reveal details of development and evolution that otherwise may have been unclear. Such details include apparent differences in the mechanisms of dorsal-ventral patterning and limb identity specification between mouse and chick and mechanistic novelties in amniotes, anurans, and urodeles. As we gain a better understanding of the details of limb development, further differences among taxa will be revealed. The use of appropriate comparative techniques in a phylogenetic context thus sheds light on evolutionary transitions in limb morphology and the generality of developmental models across species and is therefore important to both evolutionary and developmental biologists. PMID- 16246322 TI - Developmental potential and behavior of tetraploid cells in the mouse embryo. AB - Tetraploid (4n) mouse embryos die at variable developmental stages. By examining 4n embryos from F2 hybrid and outbred mice, we show that 4n developmental potential is influenced by genetic background. The imprinted inactivation of an X chromosome-linked eGFP transgene in extraembryonic tissues occurred correctly in 4n embryos. A decrease of the cleavage rate in 4n preimplantation embryos compared to diploid (2n) embryos was revealed by real-time imaging, using a histone H2b:eGFP reporter. It has previously been known that mouse chimeras produced by the combination of diploid (2n) embryos with embryonic stem (ES) cells result in mixtures of the two components in epiblast-derived tissues. In contrast, the use of 4n host embryos with ES cells restricts 4n cells from the embryonic regions of chimeras, resulting in mice that are believed to be completely ES-derived. Using H2b:eGFP transgenic mice and ES cells, the behavior of 4n cells was determined at single cell resolution in 4n:2n injection and aggregation chimeras. We found a significant contribution of 4n cells to the embryonic ectoderm at gastrulation in every chimera analyzed. We show that the transition of the embryonic regions from a chimeric tissue to a predominantly 2n tissue occurs after gastrulation and that tetraploid cells may persist to midgestation. These findings suggest that the results of previously published tetraploid complementation assays may be influenced by the presence of tetraploid cells in the otherwise diploid embryonic regions. PMID- 16246323 TI - Differential susceptibility of midbrain and spinal cord patterning to floor plate defects in the talpid2 mutant. AB - The chick talpid2 mutant displays polydactylous digits attributed to defects of the Hedgehog (HH) signaling pathway. We examined the talpid2 neural tube and show that patterning defects in the spinal cord and the midbrain are distinct from each other and from the limb. Unlike the Sonic Hedgehog (SHH) source in the limb, the SHH-rich floor plate (FP) is reduced in the talpid2 midbrain. This is accompanied by a severe depletion of medial cell populations that encounter high concentrations of SHH, an expansion of lateral cell populations that experience low concentrations of SHH and a broad deregulation of HH's principal effectors (PTC1, GLI1, GLI2, GLI3). Together with the failure of SHH misexpression to rescue the talpid2 phenotype, these results suggest that talpid2 is likely to have a tissue-autonomous, bidirectional (positive and negative) role in HH signaling that cannot be attributed to the altered expression of several newly cloned HH pathway genes (SUFU, DZIP1, DISP1, BTRC). Strikingly, FP defects in the spinal cord are accompanied by relatively normal patterning in the talpid2 mutant. We propose that this differential FP dependence may be due to the prolonged apposition of the notochord to the spinal cord, but not the midbrain during development. PMID- 16246324 TI - The role of NF-kappaB signaling in impaired liver tissue repair in thioacetamide treated type 1 diabetic rats. AB - Previously we reported that an ordinarily nonlethal dose of thioacetamide (300 mg/kg) causes liver failure and 90% mortality in type 1 diabetic rats, primarily because of inhibited tissue repair. On the other hand, the diabetic rats receiving 30 mg thioacetamide/kg exhibited equal initial liver injury and delayed tissue repair compared to nondiabetic rats receiving 300 mg thioacetamide/kg, resulting in a delay in recovery from that liver injury and survival. These data indicate that impaired tissue repair in diabetes is a dose-dependent function of diabetes. The objective of the present study was to test the hypothesis that disrupted nuclear factor-kappaB (NF-kappaB)-regulated cyclin D1 signaling may explain dose-dependent impaired tissue repair in the thioacetamide-treated diabetic rats. Administration of 300 mg thioacetamide/kg to nondiabetic rats led to sustained NF-kappaB-regulated cyclin D1 signaling, explaining prompt compensatory tissue repair and survival. For the first time, we report that NF kappaB-DNA binding is dependent on the dose of thioacetamide in the liver tissue of the diabetic rats. Administration of 300 mg thioacetamide/kg to diabetic rats inhibited NF-kappaB-regulated cyclin D1 signaling, explaining inhibited tissue repair, liver failure and death, whereas remarkably higher NF-kappaB-DNA binding but transient down regulation of cyclin D1 expression explains delayed tissue repair in the diabetic rats receiving 30 mg thioacetamide/kg. These data suggest that dose-dependent NF-kappaB-regulated cyclin D1 signaling explains inhibited versus delayed tissue repair observed in the diabetic rats receiving 300 and 30 mg thioacetamide/kg, respectively. PMID- 16246325 TI - Comparison of antibody array substrates and the use of glycerol to normalize spot morphology. AB - Antibody microarrays are a high-throughput proteomic technology used to examine the expression of multiple proteins in complex solutions. Antibody microarrays can be manufactured on a variety of commercially available activated glass or coated slides. The goal of this study was to compare Hydrogeltrade mark, nitrocellulose, aldehyde-silane and epoxy-silane slides to determine the amount of antibody bound. The optimal substrate was defined as one that bound the greatest amount of antibody with minimal background. Our studies found that epoxy silane enhanced surface (ES) slides gave the greatest degree of binding along with a minimal background. However, larger antibody microarrays showed variability in spot size, high intra-spot coefficient of variation and drying artifacts. Increasing the amount of glycerol in the spotting buffer caused a dose dependent improvement in overall spot morphology. Glycerol was tested on 128 different antibodies and showed decreased: mean spot diameter, intra-spot coefficient of variation and drying artifacts. These studies revealed that the optimal slide substrate was epoxy-silane ES microarray slides. Furthermore, glycerol could normalize spot size, decrease intra-spot coefficient of variability, decrease drying artifacts and increase antibody-spotting density. PMID- 16246326 TI - Endothelial nitric oxide synthase expression in pulmonary capillary hemangiomatosis. AB - Pulmonary capillary hemangiomatosis (PCH) is an unusual disorder characterized by the proliferation of capillaries in the alveolar septa and pulmonary interstitium. Originally conceived as a primary idiopathic disorder of the pulmonary microcirculation, recent studies have demonstrated that PCH may be associated with other pathologies. Nitric oxide (NO) is a gaseous free radical with protean biological effects that is released during the intracellular conversion of arginine to citrulline. Nitric oxide synthases (NOS) mediate the production of NO and the release of NO in the microvasculature is specifically catalyzed by endothelial NOS (NOS-III). As NOS contributes to angiogenesis and is reduced in the hypertensive pulmonary microcirculation, we examined the expression of NOS-III protein in situ in the lungs of patients with PCH. Reduced microvascular expression of NOS-III protein by endothelial cells was observed in 4/6 (67%) cases of PCH, and all of these showed concomitant pulmonary vascular hypertensive remodeling. In 2/6 (33%) cases of PCH with no morphologic evidence of pulmonary hypertensive arteriopathy, endothelial expression of NOS-III protein was judged to be either minimally reduced or normal. These findings suggest that NOS-III is specifically reduced in PCH when pulmonary arterial hypertensive remodeling is concomitantly present. PMID- 16246327 TI - The Cbl RING finger C-terminal flank controls epidermal growth factor receptor fate downstream of receptor ubiquitination. AB - Evolutionarily conserved sequences of the E3/protein-ubiquitin ligase Cbl regulate epidermal growth factor receptor (EGF-R) signaling and degradation. These sequences encompass Cbl's tyrosine kinase-binding domain, linker region, RING finger (RF), and an uncharacterized flank C-terminal to the RF (residues 420 436). The latter domain, designated the RF tail, extends beyond Cbl's ubiquitin conjugating enzyme (Ubc)-binding domain and has no known function. We report structure-function studies evaluating the impact of Cbl RF tail truncations on EGF-R fate in HEK 293 cells. All of the truncation mutants exhibit greatly reduced binding to activated EGF-R and lack proline-rich sequences that mediate direct Cbl association with SH3 proteins such as Grb2, yet a subset of mutants collectively enhances EGF-R ubiquitination, downregulation, and degradation. Significantly, EGF-R degradation correlates better with RF tail-dependent degradation of the Cbl substrate Sprouty2 than with EGF-R ubiquitination: expression of the RF tail truncation mutant Cbl 1-433 enhanced EGF-R ubiquitination while impeding Sprouty2 degradation, and Cbl 1-433 failed to enhance EGF-R downregulation or degradation. Our results suggest that EGF-R fate is controlled by a checkpoint downstream of receptor ubiquitination whose regulation by the Cbl RF tail may require Sprouty2 degradation. PMID- 16246328 TI - Long-term survival and integration of porcine expanded neural precursor cell grafts in a rat model of Parkinson's disease. AB - Porcine fetal neural tissue has been considered as an alternative source to human allografts for transplantation in neurodegenerative disorders by virtue of the fact that it can overcome the ethical and practical difficulties using human fetal neural tissue. However, primary porcine neural xenografts are rejected while porcine expanded neural precursor neural cells (PNPCs) seem to be less immunogenic and thus survive better [Armstrong, R.J., Harrower, T.P., Hurelbrink, C.B., McLaughin, M., Ratcliffe, E.L., Tyers, P., Richards, A., Dunnett, S.B., Rosser, A.E., Barker, R.A., 2001a. Porcine neural xenografts in the immunocompetent rat: immune response following grafting of expanded neural precursor cells. Neuroscience 106, 201-216]. In this study, we extended these observations to investigate the long-term survival of such transplants in immunosuppressed rats. Unilateral 6 OHDA lesioned rats received grafts into the dopamine denervated striatum of either primary porcine fetal neural tissue dissected from the E26 cortex or cortically derived neural stem cells which had been derived from the same source but expanded in vitro for 21 days. All cortically derived neural stem cell grafts survived up to 5 months in contrast to the poor survival of primary porcine xenografts. Histological analysis demonstrated good graft integration with fibers extending into the surrounding host tissue including white matter with synapse formation, and in addition there was evidence of host vascularization and myelinated fibers within the graft area. This study has therefore shown for the first time the reliable long-term survival of grafts derived from porcine expanded neural precursors in a rat model of PD, with maturation and integration into the host brain. This demonstrates that such xenografted cells may be able to recreate the damaged circuitry in PD although strategies for dopaminergic differentiation of the porcine neural precursor cell remain to be refined. PMID- 16246329 TI - Antisense oligos to neuronal nitric oxide synthase aggravate motoneuron death induced by spinal root avulsion in adult rat. AB - The present study used nitric oxide synthase (nNOS) antisense oligos (nNOS AS ODN) to assess the role of nNOS in motoneuron death induced by spinal root avulsion. A right seventh cervical (C7) spinal root avulsion was performed on adult male Sprague-Dawley rats. Two weeks later, FITC-labeled random oligos (FITC R-ODN), nNOS AS-ODN, R-ODN or TE buffer was applied to the lesioned side of the C7 spinal segment and refreshed every 3 days. FITC-R-ODN was first detected inside the injured motoneurons at 10 h, accumulated to a maximum by 24 h and faded out from 72 h. Following avulsion, nNOS AS-ODN decreased the number of nNOS positive motoneurons in the lesioned segment compared either with buffer (P < 0.001 at 15 days, 3 and 4 weeks post-injury) or with R-ODN control (P = 0.002 at 15 days, P < 0.001 at 3 and 4 weeks post-injury). Interestingly, nNOS AS-ODN also decreased the number of surviving motoneurons compared either with buffer (P = 0.005 at 15 days, P < 0.001 at 3 or 4 weeks) or with R-ODN control (P < 0.001 at 3 or 4 weeks). Meanwhile, there were no significant differences between R-ODN and buffer control either in the number of nNOS-positive motoneurons (P = 0.245 at 15 days, P = 0.089 at 3 weeks and P = 0.162 at 4 weeks) or in the number of surviving motoneurons (P = 0.426 at 15 days, P = 0.321 at 3 weeks or P = 0.344 at 4 weeks). These findings indicate that nNOS AS-ODN, applied from 2 weeks after avulsion, aggravates the motoneuron death due to root avulsion by specifically down-regulating nNOS gene expression and that the expression of nNOS in adult spinal motoneurons in response to root avulsion may play a beneficial role in the survival of injured neurons. PMID- 16246330 TI - Striatal deafferentation increases dopaminergic neurogenesis in the adult olfactory bulb. AB - Dopaminergic loss is known to be one of the major hallmarks of Parkinson disease (PD). In addition to its function as a neurotransmitter, dopamine plays significant roles in developmental and adult neurogenesis. Both dopaminergic deafferentation and stimulation modulate proliferation in the subventricular zone (SVZ)/olfactory bulb system as well as in the hippocampus. Here, we study the impact of 6-hydroxydopamine (6-OHDA) lesions to the medial forebrain bundle on proliferation and neuronal differentiation of newly generated cells in the SVZ/olfactory bulb axis in adult rats. Proliferation in the SVZ decreased significantly after dopaminergic deafferentation. However, the number of neural progenitor cells expressing the proneuronal cell fate determinant Pax-6 increased in the SVZ. Survival and quantitative cell fate analysis of newly generated cells revealed that 6-OHDA lesions induced opposite effects in the two different regions of neurogenesis in the olfactory bulb: a transient decrease in the granule cell layer contrasts to a sustained increase of newly generated neurons in the glomerular layer. These data point towards a shift in the ratio of newly generated interneurons in the olfactory bulb layers. Dopaminergic neurogenesis in the glomerular layer tripled after lesioning and consistent with this finding, the total number of tyrosine hydroxylase (TH)-positive cells increased. Thus, loss of dopaminergic input to the SVZ led to a distinct cell fate decision towards stimulation of dopaminergic neurogenesis in the olfactory bulb glomerular layer. This study supports the accumulating evidence that neurotransmitters play a crucial role in determining survival and differentiation of newly generated neurons. PMID- 16246331 TI - Apo2L/TRAIL is an indirect mediator of apoptosis induced by interferon-alpha in human myeloma cells. AB - Interferon-alpha (IFN-alpha) is currently used for the therapy of multiple myeloma (MM) though it is only effective in some patients. IFN-alpha induces apoptosis in some MM cell lines and it has been proposed to occur through an autocrine loop involving Apo2L/TRAIL. We have analysed the sensitivity to IFN alpha and Apo2L/TRAIL of five MM cell lines and found no correlation between the apoptosis inducing ability of both cytokines. IFN-alpha-induced apoptosis in MM cells was not prevented by a caspase-8 selective inhibitor (Z-IETD-fmk) or blocking Apo2L/TRAIL. However, human monocytes treated with IFN-alpha release bioactive Apo2L/TRAIL to culture media which was cytotoxic for MM cells resistant to IFN-alpha. We propose that Apo2L/TRAIL released from IFN-alpha-stimulated blood monocytes would be a major mediator of the anti-myeloma effect of IFN-alpha in vivo. PMID- 16246332 TI - Interactions of DC-SIGN with Mac-1 and CEACAM1 regulate contact between dendritic cells and neutrophils. AB - Early during infection neutrophils are the most important immune cells that are involved in killing of pathogenic bacteria and regulation of innate immune responses at the site of infection. It has become clear that neutrophils also modulate adaptive immunity through interactions with dendritic cells (DCs) that are pivotal in the induction of T cell responses. Upon activation, neutrophils release TNF-alpha and induce maturation of DCs that enables these antigen presenting cells to stimulate T cell proliferation and to induce T helper 1 polarization. DC maturation by neutrophils also requires cellular interactions that are mediated by binding of the DC-specific receptor DC-SIGN to Mac-1 on the neutrophil. Here, we demonstrate that also CEACAM1 is an important ligand for DC SIGN on neutrophils. Binding of DC-SIGN to both CEACAM1 and Mac-1 is required to establish cellular interactions with neutrophils. DC-SIGN is a C-type lectin that has specificity for Lewis(x), and we show that DC-SIGN mediates binding to CEACAM1 through Lewis(x) moieties that are specifically expressed on CEACAM1 derived from neutrophils. This indicates that glycosylation-driven binding of both Mac-1 and CEACAM1 to DC-SIGN is essential for interactions of neutrophils with DCs and enables neutrophils to modulate T cell responses through interactions with DCs. PMID- 16246333 TI - Malaria parasite-infected erythrocytes inhibit glucose utilization in uninfected red cells. AB - The erythrocytic stages of the malaria parasite depend on anaerobic glycolysis for energy. Using [2-13C]glucose and nuclear magnetic resonance, the glucose utilization rate and 2,3-diphosphoglycerate (2,3-DPG) level produced in normal RBCs and Plasmodium falciparum infected red blood cell populations (IRBCs, with <4% parasite infected red cells), were measured. The glucose flux in IRBCs was several-folds greater, was proportional to parasitemia, and maximal at trophozoite stage. The 2,3-DPG levels were disproportionately lower in IRBCs, indicating a downregulation of 2,3-DPG flux in non-parasitized RBCs. This may be due to lowered pH leading to selective differential inhibition of the regulatory glycolytic enzyme phosphofructokinase. This downregulation of the glucose utilization rate in the majority (>96%) of uninfected RBCs in an IRBC population may have physiological implications in malaria patients. PMID- 16246335 TI - Interaction of fraternal birth order and handedness in the development of male homosexuality. AB - The present study investigated evidence for an interaction between two of the best established etiologic factors, or markers of etiologic factors, in the literature on male homosexuality: fraternal birth order and hand preference. By combining five samples, the authors produced study groups of 1774 right-handed heterosexuals, 287 non-right-handed heterosexuals, 928 right-handed homosexuals, and 157 non-right-handed homosexuals. The results showed a significant (P = 0.004) handedness by older brothers interaction, such that (a) the typical positive correlation between homosexuality and greater numbers of older brothers holds only for right-handed males, (b) among men with no older brothers, homosexuals are more likely to be non-right-handed than heterosexuals; among men with one or more older brothers, homosexuals are less likely to be non-right handed than heterosexuals, and (c) the odds of homosexuality are higher for men who have a non-right hand preference or who have older brothers, relative to men with neither of these features, but the odds for men with both features are similar to the odds for men with neither. These findings have at least two possible explanations: (a) the etiologic factors associated with non-right handedness and older brothers-hypothesized to be hyperandrogenization and anti male antibodies, respectively-counteract each other, yielding the functional equivalent of typical masculinization, and (b) the number of non-right-handed homosexuals with older brothers is smaller than expected because the combination of the older brothers factor with the non-right-handedness factor is toxic enough to lower the probability that the affected fetus will survive. PMID- 16246334 TI - Effects of ACTH and cAMP on steroidogenic acute regulatory protein and P450 11beta-hydroxylase messenger RNAs in rainbow trout interrenal cells: relationship with in vitro cortisol production. AB - Steroidogenic acute regulatory protein (StAR) transfers cholesterol over the inner mitochondrial membrane, thereby making the molecule available for cholesterol side-chain cleavage enzyme, which carries out the first conversion in the steroidogenic pathway. In mammals, StAR controls this rate limiting step in steroidogenesis, and both StAR protein and StAR mRNA levels become rapidly elevated in response to tropic hormone stimulation. The relationship between StAR gene expression and steroid production in fish has not yet been well explored. We investigated the relationship between adrenocorticotropic hormone (ACTH)- and cAMP-stimulated cortisol production in vitro and levels of StAR transcripts in interrenal cells of rainbow trout. To assess the effect of ACTH on mRNA levels of a downstream steroidogenic enzyme, we also investigated the effects of ACTH on transcripts encoding 11beta hydroxylase (P450 11beta). In a series of experiments, juvenile rainbow trout head kidney tissue containing interrenal cells was incubated with either ACTH or dibutyryl cyclic AMP (dbcAMP). Cortisol in incubation media were measured by radioimmunoassay and total RNA was isolated from the tissue for Northern analysis or for quantitative real-time PCR. Incubation of tissue with 150 ng/mL ACTH for 1-18 h induced a progressive increase in cortisol accumulation in media, but StAR mRNA levels increased modestly and mostly insignificantly over 18 h, irrespective of treatment. Exposure of tissue for 18 h to 5, 150, 500 or 1,500 ng ACTH/mL resulted in a strong increase in cortisol production, with a peak response (15-fold increase over controls) achieved with 150 ng/mL ACTH. Although there was a trend towards a dose-response effect, mean StAR mRNA levels were only significantly affected by the highest concentration of ACTH used (1,500 ng/mL), which induced a less than 2 fold increase in StAR transcripts. However, there was a significant linear relationship between StAR mRNA levels and ACTH-induced cortisol accumulation in media (p<0.001, r(2)=0.55). Incubation of tissue with 5mM dbcAMP for 6 or 18 h induced large increases in cortisol accumulation in media over controls, but had no significant effect on StAR mRNA levels. By contrast, ACTH induced a clear dose dependent increase in P450 11beta transcripts, with 150 ng/mL ACTH inducing an 8 fold increase in levels compared to control; nonetheless, only a weak correlation existed between transcript levels and ACTH-induced cortisol secretion (p<0.003, r(2)=0.26). Thus, despite the relatively high degree of conservation of StAR proteins in vertebrates, we have been unable to demonstrate that a rapid, acute increase in transcription of the StAR gene is the dominant mechanism supporting flow of cholesterol to the mitochondria during acute increases in cortisol production in rainbow trout. The strong stimulation of P450 11beta gene transcription by ACTH probably enhances biosynthetic capacity during longer term chronic ACTH stimulation. PMID- 16246336 TI - Fatal cardiac air embolism. PMID- 16246337 TI - Reduced time on the spinal board-effects of guidelines and education for emergency department staff. AB - AIM: Prehospital spinal immobilisation is usually accomplished with a spinal board. Prolonged immobilisation on spinal boards in the emergency department (ED) can be detrimental. This study aimed to reduce the time spent by patients on spinal boards using a staff education program. METHODS: Observational study in a trauma centre ED seeing 180,000ED attendances per year. The length of time immobilised on spinal board was recorded by the trauma nurse coordinator. Guidelines on removal of spinal boards were issued after recording period 1 (January-June 2001) and reinforced several times. The post-training period (period 2) extended from May to October 2003. Medians were compared using Mann Whitney U-test (non-parametric data); chi-square test was used for categorical data. RESULTS: There were 122 eligible patients in period 1 and 104 eligible patients in period 2. Median time to removal from the spinal board was reduced by 18.5 min from 50 to 31.5 min (Mann-Whitney U-test, p<0.0001, 95% CI for difference in medians 13-29 min). In period 1, 44 of 122 patients (36%) were removed from the spinal board before leaving the ED, compared to 78 of 104 patients (75%) in period 2 (p<0.0001, chi-square test). CONCLUSION: The introduction of guidelines, reinforced by ED staff education, can significantly reduce the time patients spend on spinal boards after trauma and can increase the proportion of patients who can be removed from the board before leaving the ED. PMID- 16246338 TI - Liver injuries in children: the role of selective non-operative management. AB - INTRODUCTION: This review article on the management of blunt liver injury in children is based on the authors' experience of 311 patients over a 22-year period. MATERIAL AND METHODS: All children presenting to our institution with confirmed blunt liver trauma were studied retrospectively. Hospital folders of 311 patients were analysed. Information was gathered about the clinical presentation, associated injuries, grade of injury, transfusion requirements and haemodynamic stability to examine factors influencing outcome. RESULTS: The age of patients ranged between 3 weeks and 12 years (mean of 7 years). Injuries as a result of motor vehicle accidents (MVAs) were the most common (268; 232 pedestrian and 36 passenger), other causes were falls (26) assaults or child abuse(15), bicycle handle bar injury (2). One hundred and thirty-six patients sustained an isolated hepatic injury and 175 had multiple injuries. Associated injuries included 147 head injuries, 131 fractures, 66 thoracic and 143 intra abdominal (74 spleen, 45 renal, 4 pancreatic and 4 hollow viscus). Two patients died soon after arrival, 21 underwent laparotomy, 13 of which were liver related, while 288 were treated non-operatively. One hundred and six patients required blood transfusion (mean of 21.3 ml/kg); 30% of the nonoperative group and 100% of the operative group. There were three fatalities from the operative group (1% total mortality), one secondary to a severe, head injury, one liver haemorrhage and one from multi-organ failure DISCUSSION: The vast majority (93%) was successfully treated non-operatively with only 4% coming to liver related laparotomy, complications were lower, transfusions less and the in-hospital occupancy was shorter. Complication rate was 8% and mortality was 1%. CONCLUSION: We confirm the success selective non-operative management of blunt liver trauma as adopted by this institution 20 years ago. It is now proven treatment in an appropriate centre. However, the challenge is to identify the severely injured child early and institute aggressive resuscitation and expedite laparotomy when indicated. PMID- 16246339 TI - Non-reducible, open tibial plafond fractures treated with a circular external fixator (is the current classification sufficient for identifying fractures in this area?). AB - While some researchers advocate primary arthrodesis for the treatment of open, severely comminuted tibial plafond fractures, others argue that an external fixator is an alternative. In this study, we obtained intermediate clinical and radiological outcomes on treating such fractures with a circular external fixator. Twelve patients with non-reducible, open tibial plafond fractures were treated with a circular external fixator and minimal osteo-synthesis. The fractures were grouped using a modification of the Ruedi and Allgower classification: eight, two, and two of the patients had Type III, IVA, and IVB fractures, respectively. The bone transport technique was applied in the patients with a Type IVB fracture. Four parameters were tracked in the patients: the reduction score of the joint surface, early complications, and the radiological and clinical findings of the ankle. The average follow-up period of the patients was 54.5 months. In the postoperative radiological examinations, the reduction score of the joint surface exceeded 15 in four patients and was 12-15 in eight patients. Type III and IVA fractures united with an average healing time of 4.25 months. Surface wound infection was observed in three patients. One patient each developed fibular osteomyelitis, claw toe, and 2cm shortness. Among the patients with Type IVB fractures, nonunion and malunion in the newly forming callus was observed in one patient, and nonunion alone was observed in another patient. On the final check, both the clinical and radiological findings were poor for all of the patients, with a reduction score exceeding 15. High-energy and poor joint surface reduction scores are two important factors affecting both the clinical and radiological results. For the fractures with reduction scores below 15, it is particularly difficult to predict the clinical results. Therefore, we recommend that such fractures be treated with a circular external fixator and believe that arthrodesis in accordance with the patient's choice is a desirable treatment method. PMID- 16246340 TI - Plaster cast immobilisation during air travel-analysis of current practice and aircraft-simulated experimental study. AB - BACKGROUND: There is an increase in aircraft transportation of patients with lower limb fractures. Current practice is variable. Our aim was to study current practice and to analyse the situation in an experimental simulated aircraft flight. METHODS: Current advice supplied by commercial airline offices in the UK was noted. Postal questionnaires were sent to Orthopaedic Consultants in the UK to obtain their current practice. Experimental aircraft travel was simulated in a decompression chamber with five medically fit volunteers with no fracture, immobilised in an above knee plaster cast. Compartment pressure and venous return was documented and the results analysed in two different positions with the leg elevated and dependant. RESULTS: Airlines do not have any formal guidelines. Orthopaedic consultants in the UK note variable advise in this situation. Experimental study in the aircraft simulation showed that two volunteers developed significant increase in compartmental pressure with the leg elevated to 90 degrees , which settled after the plaster cast was split. There was no increase in compartment pressure noted with leg dependant on the floor with 45 degrees of flexion at hip. CONCLUSION: The literature on this issue is limited. With our analysis we feel that patients can be transported with the plaster cast split with limb dependant on the floor i.e. hip flexion less than 45 degrees . Our volunteers had no fractures so direct comparison with pathological changes in acute fracture is problematical. Further studies into this problem are recommended. PMID- 16246341 TI - Avulsion fractures of the lateral tibial condyle in children. AB - Avulsion fractures of the lateral tibial condyle are very rare in children and adolescents. Seven patients were treated between 1986 and 2000. They were all involved in sporting activities and an acute contraction of the fibres of the iliotibial band was considered to be their common pathogenetic factor. A subdivision in two types of avulsion fractures according to the anatomic location of the lesion was used. The Segond fracture, a small vertical avulsion fracture involving the midportion of the lateral tibial condyle distal to the plateau but proximal to the physis, was diagnosed in five patients. In two of them it was associated with a fracture of the anterior intercondylar eminence. The avulsion fracture of the Gerdy's tubercle was diagnosed in two other patients. The detached fragment was bigger, more anterior and involved a part of the articular surface of the lateral tibial plateau as well as a part of the physeal plate. All patients were treated conservatively and were free of symptoms after a mean follow-up of 9 years. PMID- 16246342 TI - Cervical ripening and induction of labor with intravaginal misoprostol and Foley catheter cervical traction. PMID- 16246343 TI - Type-specific distribution of human papillomavirus in relation to grades of cervical neoplasia. PMID- 16246344 TI - Delivery-related complications and early postpartum morbidity in Dhaka, Bangladesh. AB - OBJECTIVE: To describe delivery-related complications and postpartum morbidity of women living in slum areas of Dhaka, Bangladesh. METHOD: From November 1993 to May 1995, 1506 women were interviewed regarding delivery-related complications and postpartum morbidities. Operational definitions were applied to maternal reports to categorize serious delivery-related complications and postpartum morbidity. Corroborating information was identified from medical records for facility-based deliveries and physical examinations by female physicians 14 to 22 days postpartum. RESULT: Thirty-six percent of women described serious delivery related complications and 75% of women reported postpartum morbidity. There were two maternal deaths among 1471 live births. When maternal reports were related to corroborating information, the proportion of women's reports of serious complications and morbidity appears reasonably accurate for some conditions. CONCLUSION: A large proportion of urban slum women in Dhaka experience serious delivery-related complications and/or postpartum morbidity. Information on delivery practices that contribute to morbidity and factors that influence appropriate care seeking is needed. PMID- 16246345 TI - Cocaine intoxication mimicking preeclampsia postpartum. PMID- 16246346 TI - 3-Hydroxyanthranilic acid, one of L-tryptophan metabolites, inhibits monocyte chemoattractant protein-1 secretion and vascular cell adhesion molecule-1 expression via heme oxygenase-1 induction in human umbilical vein endothelial cells. AB - Heme oxygenase (HO)-1 is important in the vascular system, and its genetic or pharmacological induction in endothelium would be effective for the prevention and treatment of atherosclerosis. The naturally occurring antioxidant 3 hydroxyanthranilic acid (HA), one of l-tryptophan metabolites formed in vivo along the metabolic route known as the kynurenine pathway during inflammation or infection, was found to induce HO-1 expression and to stimulate nuclear translocation of NF-E2 related factor 2 (Nrf2) in human umbilical vein endothelial cells (HUVECs). Pre-treatment with HA inhibited the secretion of monocyte chemoattractant protein (MCP)-1, the expression of vascular cell adhesion molecule (VCAM)-1 and the activation of transcriptional nuclear factor (NF)-kappaB in HUVECs stimulated with tumor necrosis factor-alpha, the major pro inflammatory cytokine causing endothelial inflammation. Interestingly, the observed anti-inflammatory effects of HA were mimicked by a HO-1 inducer, cobalt protoporphyrin, and bilirubin, one of HO-1 enzymatic products, but abolished in the presence of a HO-1 inhibitor, tin protoporphyrin. Based on our findings, we suggest that Nrf2-dependent HO-1 expression induced by HA inhibits MCP-1 secretion, VCAM-1 expression and NF-kappaB activation associated with vascular injury and inflammation in atherosclerosis. PMID- 16246348 TI - Quantitative determination of chlortetracycline content in animal plasma at controlled keto-enol tautomerism by liquid chromatography-electrospray ionization tandem mass spectrometry. AB - Chlortetracycline (CTC) is a member of the group of the tetracycline antibiotics used in human and veterinary medicine. Its reliable measurement in plasma - necessary to study its pharmacokinetics - is hampered by the fact that CTC is subjected to keto-enol tautomerism resulting in its keto- and enol-form. Also, it undergoes epimerization to form its 4-epimer. In this paper, we present an LC-ESI MS/MS method to analyze the CTC content in animal plasma, with the originality that it apports a solution to overcome the possible interference of the keto-enol tautomerism equilibrium, which is influenced by several factors. Therefore, the plasma samples were incubated at 56 degrees C, forcing the equilibrium completely in favour of the CTC keto-form. The CTC keto-tautomer was stable after deproteinization of the samples with concentrated trifluoroacetic acid, and was quantified by LC-ESI-MS/MS, as a measure of the total CTC content. The method could be validated for pig and chicken plasma by a series of parameters - linearity, trueness, precision, limit of quantification, specificity, stability - in accordance with the European guidelines, and was used for pharmacokinetic studies on pigs and chickens medicated with CTC via the feed. PMID- 16246347 TI - Ethnic differences of the presence and severity of coronary atherosclerosis. AB - BACKGROUND: Although cardiovascular risk factor levels are substantially different in Caucasians, African-American, Hispanics, and Asians, the relative rates of coronary heart disease in these groups are not consistent with these differences. The objective of the study is to assess the differences in the prevalence and severity of coronary artery calcification, as a measure of atherosclerosis, in these different ethnic groups. METHODS: Electron-beam tomography was performed in 16,560 asymptomatic men and women (Asians=1336, African-Americans=610, Hispanics=1256) aged >or=35 years referred by their physician for cardiovascular risk evaluation. The study population encompassed 70% males, aged 52+/-8 years. RESULTS: Caucasians were more likely to present with dyslipidemia (p<0.0001), while African-Americans and Hispanics had a higher prevalence of smoking, diabetes, and hypertension (all p<0.001). After adjustment for age, gender, risk factors, and treatment for hypercholesterolemia, compared with Caucasians, the relative risks for men having coronary calcification were 0.64 (95% CI: 0.48-0.86) in African-Americans, 0.88 (95% CI: 0.67-1.15) in Hispanics, and 0.66 (95% CI: 0.55-0.80) in Asians. After similar adjustments, the relative risks for women having coronary calcification, were 1.58 (95% CI: 1.13 2.19) for African-Americans, 0.84 (95% CI: 0.66-1.06) in Hispanics, and 0.71 (95% CI: 0.56-0.89) in Asian women. After adjusting for age and risk factors using multivariable analysis, African-American men were least likely to have any coronary calcium while African-American women had significantly higher OR of any calcification. Asian men and women had significantly lower OR of any calcification. There was no significant difference in prevalence or severity of atherosclerosis between Hispanics and Caucasians, in men or women. CONCLUSIONS: Our study results demonstrate significant difference in the presence as well as severity of calcification according to ethnicity, independent of atherosclerotic risk factors. Results from this study (physician referred) closely parallel the results from MESA (population based, measured risk factors). Ethnic specific data on the predictive value of differing coronary calcium scores are needed. PMID- 16246349 TI - Analytical and semipreparative high performance liquid chromatography enantioseparation of new substituted 1-thiocarbamoyl-3,5-diaryl-4,5-dihydro-(1H) pyrazoles on polysaccharide-based chiral stationary phases in normal-phase, polar organic and reversed-phase conditions. AB - The direct HPLC enantioseparation of five pairs of new chiral pyrazole derivatives on coated cellulose- and amylose-based chiral stationary phases (Chiralpak AD, Chiralcel OJ and Chiralcel OJ-RH) and new immobilised amylose based Chiralpak IA CSP was performed. Very high enantioselectivity factor (alpha) values were achieved in polar organic and reversed-phase conditions by using OJ RH as CSP. Chiralpak IA exhibited an excellent chiral resolving ability in normal phase mode and it allowed the enantioseparation of analytes investigated with resolution factors (Rs) >20. Due to its bonded nature, it was successfully employed at analytical and semipreparative scale in combination with normal-phase eluents containing "non-standards" solvents such as acetone. PMID- 16246350 TI - Immobilized versus coated amylose tris(3,5-dimethylphenylcarbamate) chiral stationary phases for the enantioselective separation of cyclopropane derivatives by liquid chromatography. AB - The solvent versatility of Chiralpak IA, a new chiral stationary phase (CSP) containing amylose tris(3,5-dimethylphenylcarabamate) immobilized onto silica gel, is investigated for the enantioselective separation of a set of cyclopropane derivatives using ethyl acetate or dichloromethane (DCM) as non-standard mobile phase eluent and diluent, respectively in high-performance liquid chromatography (HPLC). A comparison of the separation of cyclopropanes on both immobilized and coated amylose tris(3,5-dimethylphenylcarbamate) chiral stationary phases (Chiralpak IA and Chiralpak AD, respectively) in HPLC using a mixture of n hexane/2-propanol (90/10 and 99/1, v/v) as mobile phase with a flow rate of 0.5 ml/min and UV detection at 254 nm, is demonstrated. The optimized method of separation is used for an online HPLC monitoring for the Rh(II)-catalyzed asymmetric intermolecular cyclopropanations in dichloromethane. Direct analysis techniques without further purification, workup or removal of dichloromethane were summarized. The method provides an easy and direct determination of the enantiomeric excess of the cyclopropanes and selectivity of the catalyst used without any further work up. PMID- 16246351 TI - Characterization of waxes used in pictorial artworks according to their relative amount of fatty acids and hydrocarbons by gas chromatography. AB - A study attempted to characterize natural waxes used in pictorial works of art was carried out by means of gas chromatography. The analytical treatment requires prior hydrolysis of the waxes to release the fatty acids (FA) (myristic (myr), palmitic (pal), oleic (ole), stearic (ste), araquidic (ara), behenic (beh), lignoceric (lig), cerotic (cer)) from the main esters of the waxes. The formation of volatile derivatives of the fatty acids was carried out by derivatization with ethyl choroformate (ECF). This derivatization reagent was chosen due to the speed, safety and quantitativity of the reaction. The analyzed hydrocarbons were n-eicosane, n-heneicosane, n-docosane, n-tricosane, n-tetracosane, n-pentacosane, n-hexacosane, n-heptacosane, n-octacosane, n-nonacosane, n-tricontane n hentriacontane, n-dotriacontane, n-tritriacontane, n-tetratriacontane, n pentatriacontane, main constituents of the waxes. No derivatization is needed to analyze the hydrocarbons. Ethyl ester derivatives and hydrocarbons are adequately separated by gas chromatography, identified by flame ionization detection and confirmed by mass spectrometry. To characterize natural waxes, peak area ratios of each fatty acids with respect to the palmitic acid and peak area ratios of each hydrocarbons with respect to n-heptacosane were calculated. The proposed method provides a good characterization of different waxes most frequently used in artworks, such as beeswax, carnauba wax and ceresin, and has been successfully applied to real samples. This is the first report on the application of ECF to the analysis of fatty acids in wax. PMID- 16246352 TI - Identification of isomers and subsidiary colors in commercial Fast Green FCF (FD&C Green No. 3, Food Green No. 3) by liquid chromatography-mass spectrometry and comparison between amounts of the subsidiary colors by high-performance liquid chromatography and thin-layer chromatography-spectrophotometry. AB - The existence of five of the six expected isomers in commercial Fast Green FCF (G3: Food Green No. 3, FD&C Green No. 3, CAS No. 2353-45-9, C.I. No. 42053), the main product of which is m,m-G3 and the sub-products of which are presumed to be m,p-G3, o,m-G3, p,p-G3, o,p-G3 and o,o-G3, was confirmed using LC/MS, and the levels of the isomers, m,m-G3, m,p-G3, p,p-G3, o,m-G3 and o,p-G3, were determined by analytical HPLC. The existence of seven subsidiary colors that were decomposed from G3 was also confirmed using LC/MS. The levels of the subsidiary colors in ethanol extracts from TLC were determined by HPLC and spectrophotometry, and these results were compared. It was clear that the values determined by TLC spectrophotometry were higher than those by HPLC. It was recommended that the levels of subsidiary colors in G3 should be determined by HPLC. PMID- 16246353 TI - Headspace solid-phase microextraction gas chromatography-mass spectrometry method for the identification of cosmetic ingredients causing delamination of packagings. AB - A headspace solid-phase microextraction gas chromatography-mass spectrometry (HS SPME-GC-MS) method using a 75 microm Carboxen polydimethylsiloxane fibre was used to identify volatile compounds of cosmetic formulations responsible for causing loss of adhesion between layers of multilayer packagings. To obtain the sample, the sachet with the product was kept in an oven at 40 degrees C in order to favour the migration of the aggressive compounds to the inner layers. Then the sachet was manually delaminated and the aluminium/polyester and polyethylene layers were analysed. The cosmetic product was also analysed by HS-SPME-GC-MS. Several compounds used in the cosmetic industry such as perfumes or fixing agents were detected in the inner layers of the laminated material, showing the migration of them through the layer in contact with the product (polyethylene). Phenoxy ethanol, beta-linalool, menthol and p-propenylanisole are suspected to be responsible for the loss of adhesion. In order to provide a complete overview of the cause of the aforementioned phenomenon, the packaging material was exposed to the cosmetic products in order to measure the decrease of the adhesion strength with time. It was observed that the product with a higher phenoxy ethanol concentration caused a higher loss of adhesion strength. The results obtained showed that this method is suitable for identifying aggressive compounds in cosmetic products, as well as for giving prior information about which products may be problematic for packaging in sachets. PMID- 16246354 TI - Evaluating the bioavailability of explosive metabolites, hexahydro-1-nitroso-3,5 dinitro-1,3,5-triazine (MNX) and hexahydro-1,3,5-trinitroso-1,3,5-triazine (TNX), in soils using passive sampling devices. AB - The uptake kinetics of two major RDX (hexahydro-1,3,5-trinitro-1,3,5 triazacyclohexane) metabolites, hexahydro-1-nitroso-3,5-dinitro-1,3,5-triazine (MNX) and hexahydro-1,3,5-trinitroso-1,3,5-triazine (TNX), into passive sampling devices (PSDs), and the ability of PSDs to serve as surrogates for evaluating bioavailability of MNX and TNX were investigated in laboratory sand and two soil types. The results indicate that MNX and TNX absorption into PSDs was best fitted with a polynomial curve model: y = ax2 + bx + c (y: amount of MNX or TNX absorbed into PSD; x: incubation time of PSDs in soil), with an excellent correlation coefficient (>0.95) for each type of soil amended with 10 mg/kg MNX or TNX. TNX was more readily absorbed by PSDs than MNX. Soil conditions, especially organic matter content, affected MNX and TNX uptake into PSDs. A relatively good correlation between MNX and TNX uptake into PSDs and uptake into earthworms was obtained in two types of natural soils (a silt loam soil from Nebraska and a sandy loam soil from Texas) and laboratory sand. A linear relationship between PSD uptake and earthworm uptake was observed. The correlation coefficients (r2) were > or = 0.82 for all test soils spiked with MNX or TNX. Organic matter content is one soil factor that affected the ratio of MNX or TNX uptake into earthworms versus uptake into PSDs. These data indicate that C18 PSDs may be used as a surrogate for soil organisms such as earthworms and provide a simple and easy chemical test for assessing the bioavailability of contaminants in soils. PMID- 16246355 TI - Analysis of triacylglycerols in fat body of bumblebees by chromatographic methods. AB - Triacylglycerols (TAGs) from the fat body of several bumblebee species (Bombus lucorum, B. terrestris, B. lapidarius, B. hypnorum, B. hortorum, and B. confusus) were studied using chromatographic techniques. Semi-preparative thin-layer chromatography was used to isolate the TAGs from the tissue extract. Gas chromatography (GC) enabled us to identify the fatty acids (FAs) that form bumblebee TAGs and to quantify their relative proportions. The TAGs were subsequently analysed by high-performance liquid chromatography-atmospheric pressure chemical ionisation mass spectrometry. Two chromatographic systems, including non-aqueous reversed-phase chromatography and silver ion chromatography on cation exchange resin in silver (I) ionic form, were optimised and their performance compared. The most abundant fatty acids in bumblebees TAGs contained 18 or 16 carbon atoms; oleic acid predominated in most samples. TAGs were found to be a complex mixture of isomers; some of them, e.g. OLnO, PLnO, PoPoO, PoPoP, POO, or OOO (where Po is palmitoleic, P is palmitic, Ln is linolenic, and O is oleic acid) were abundant in particular species. The composition of both FAs and TAGs was found to be species-specific. Only minor differences were found among specimens of the same species. PMID- 16246356 TI - Effect of embedded metal compound on porosity of silica colloids prepared by spray reaction of silicon tetrachloride. AB - Attempts to prepare macroporous silica particles and metal-compound-nanoparticle embedded silica microspheres were carried out using reactions between silicon tetrachloride and ultrasonic generating microdroplets including metal (Na, K, Al, Ni, Ti, Pt) compounds. Samples were collected by dry and wet processes. In the case of using nickel and aluminum compounds, acid-treated samples were also prepared. The obtained samples were characterized by scanning electron microscopy, X-ray fluorescence spectroscopy, powder X-ray diffractometry, mercury porosimetry, and the nitrogen adsorption method. The macroporous silica particles were prepared by removing the salt crystals, such as NaCl and KCl, formed in the silica frame. For acid-resistant metals, platinum- and titanium-compound nanoparticles are easily embedded in silica microspheres using these metal compound solutions. For acid-soluble metals, aluminum- and nickel-compound nanoparticle-embedded silicas were prepared by applying neutralization of the collection water. Micropores and mesopores were produced in wet-process samples. Acid treatment induced the increase of micropore volumes. PMID- 16246357 TI - Rheology and UV protection properties of suspensions of fine titanium dioxides in a silicone oil. AB - Ultrafine particles of titanium dioxide (TiO2) are very attractive as a UV protection ingredient in cosmetic products. The UV-scattering behavior of TiO2 suspensions in a silicone oil are studied in relation to rheological properties. To control the dispersion stability of suspensions, two types of polyether modified silicones are used as dispersants. When the suspensions are prepared with branch-type dispersants in which the polyether groups are incorporated as side chains along the backbone, the flow is shear-thinning even at low shear rates. The appearance of plateaus in the frequency-dependence curves of storage modulus implies the solidlike responses. On the other hand, the suspensions prepared with linear conformation dispersants, in which the silicone group and polyether group are alternately repeated in one long chain, are Newtonian at low shear rates. The suspensions are regarded as liquids, because the storage modulus decreases rapidly in the low-frequency region. The suspension rheology is strongly associated with flocculated structures that are primarily controlled by the interparticle attractions. The differences in rheological behavior can be explained by the differences in the adsorbed conformation of dispersant silicones. From optical measurements, it is confirmed that UV scattering increases with decreasing flocculation degree. Therefore, good agreement is established between rheological properties and UV protection ability. PMID- 16246358 TI - The role of phonological recoding in orthographic learning. AB - The objective of this research was to explore whether orthographic learning occurs as a result of phonological recoding, as expected from the self-teaching hypothesis. The participants were 32 fourth- and fifth-graders (mean age = 10 years 0 months, SD = 7 months) who performed lexical decisions for monosyllabic real words and pseudowords under two matched experimental conditions: a read aloud condition, wherein items were named prior to lexical decision to promote phonological recoding, and a concurrent articulation condition, presumed to attenuate phonological recoding. Later, orthographic learning of the pseudowords was evaluated using orthographic choice, spelling, and naming tasks. Consistent with the self-teaching hypothesis, targets learned with phonological recoding in the read aloud condition yielded greater orthographic learning than those learned with concurrent articulation. The research confirms the critical nature of phonological recoding in the development of visual word recognition skills and an orthographic lexicon. PMID- 16246359 TI - A differential assay of NK-cell-mediated cytotoxicity in K562 cells revealing three sequential membrane impairment steps using three-color flow-cytometry. AB - Annexin V and propidium iodide (PI) staining is a general technique for detecting apoptosis by flow-cytometry (FCM). The release of 2',7'-bis-(2-carboxyethyl)-5- (and-6)-carboxyfluorescein (BCECF), a non-lipophilic membrane-impermeable labeling dye, from the cytoplasm of target cells is an indicator of increased membrane permeability. This study aimed to devise a three-color FCM technique involving the BCECF-release parameter in addition to conventional Annexin V and PI staining for the analysis of target K562 cells undergoing cytotoxic/apoptotic processes mediated by natural killer (NK) cells. The results demonstrated the following step-wise process of membrane impairment: (1) initiation of Annexin V staining accompanied by increasing forward scatter (FSC) before BCECF-release, indicating membrane impairment without permeabilization by necrosis; (2) BCECF release with decreasing FSC before PI influx; and (3) PI staining with the lowest FSC state. Therefore, the early stage of cytotoxicity/apoptosis conventionally defined by the flow-cytometric criteria of Annexin V staining before PI staining could be sub-divided into two stages before and after BCECF-release. Annexin-V staining in tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) induced apoptosis was also initiated without BCECF-release. Although the underlying mechanism of the transition process from stage 1 to stage 2 is still unknown, this FCM technique should be a useful tool for differential assays of target cells regarding the sequential processes of NK-induced cytotoxicity. PMID- 16246360 TI - Environmental physiology of three species of Collembola at Cape Hallett, North Victoria Land, Antarctica. AB - The environmental physiology of three speciesof Collembola: Cryptopygus cisantarcticus, Isotoma klovstadi (Isotomidae) and Friesea grisea (Neanuridae) was investigated from November 2002 to February 2003 at Cape Hallett, North Victoria Land, Antarctica. All three species were freeze avoiding, and while supercooling points were variable on seasonal and daily scales in I. klovstadi and C. cisantarcticus, they remained largely static in F. grisea. LT50 (temperature where 50% of animals are killed by cold) was -13.6, -19.1 and -19.8 degrees C for C. cisantarcticus, I. klovstadi and F. grisea, respectively. Upper lethal temperature was 34, 34 and 38 degrees C for C. cisantarcticus, I. klovstadi and F. grisea. Critical thermal minimum onset (the temperature where individuals entered chill coma) was ca. -7, -12 and -8 degrees C for C. cisantarcticus, I. klovstadi and F. grisea, and 25% of I. klovstadi individuals froze without entering chill coma. Critical thermal maximum (the onset of spasms at high temperature) was 30, 33 and 34 degrees C for C. cisantarcticus, I. klovstadi and F. grisea. Haemolymph osmolality was approximately 720 mOsm for C. cisantarcticus and 680 mOsm for I. klovstadi, and both species showed a moderate degree of thermal hysteresis, which persisted through the season. Desiccation resistance was measured as survival above silica gel, and the species survived in the rank order of C. cisantarcticus<< I. klovstadi = F. grisea. Desiccation resulted in an increase in haemolymph osmolality in I. klovstadi, and water was quickly regained by desiccation-stressed individuals that had access to liquid water, but not by individuals placed in high humidity, indicating that this species is unable to absorb atmospheric water vapour. SDS-PAGE did not suggest any strong patterns in protein synthesis either seasonally or in response to temperature or desiccation stress. Microclimate temperatures were measured at sites representative of collection sites for the three species. Microclimate temperatures were highly variable on a diurnal and weekly scale (the latter relating to weather patterns), but showed little overall variation across the summer season. Potentially lethal high and low temperatures were recorded at several sites, and it is suggested that these temperature extremes account for the observed restriction of the less-tolerant C. cisantarcticus at Cape Hallett. Together, these data significantly increase the current knowledge of the environmental physiology of Antarctic Collembola. PMID- 16246362 TI - Temperature softening of a protein in single-molecule experiments. AB - Mechanical flexibility is crucial for the function of proteins. However, such material properties are not easily accessible experimentally. We used single molecule force spectroscopy to study the stiffness of a single domain of Dictyostelium discoideum filamin (ddFLN4) in a temperature range from 5 degrees C to 37 degrees C. Analyzing the distributions of unfolding forces allowed us to extract transition barrier heights and positions of the underlying energy landscape. We found a marked narrowing of unfolding force distributions with increasing temperature. This narrowing reflects an increase in transition state position from 2.7 A to 7.8 A and thus a reduction of the molecular spring constant of the protein by a factor of 7. We suggest this temperature softening reflects a shift in the nature of the interactions responsible for mechanical stability from hydrogen bonds to hydrophobic interactions. This result has important consequences for all interpretations of protein mechanical studies if experimental results obtained at room temperature are to be transferred to physiological temperatures. PMID- 16246361 TI - Platelets activated by collagen through the immunoreceptor tyrosine-based activation motif in the Fc receptor gamma-chain play a pivotal role in the development of myocardial ischemia-reperfusion injury. AB - Platelet activation and the formation of platelet microaggregates in coronary vessels play pivotal roles in myocardial ischemia and reperfusion injury. The Fc receptor gamma-chain (FcR gamma) is coexpressed with glycoprotein (GP) VI, forming a platelet collagen receptor, and the activation of platelets by collagen is closely coupled with tyrosine phosphorylation of the FcRgamma. To examine the functional significance of platelet FcR gamma/GPVI complex in the early phase of myocardial ischemia and reperfusion injury in mice, we performed coronary occlusion and reperfusion experiments using wild type mice and FcRgamma-deficient (FcRgamma(-/-)) mice that lack GPVI. The infarct size was significantly smaller in FcRgamma(-/-) mice subjected to occlusion and reperfusion of the coronary artery than in control FcR gamma(+/+) mice. Twenty-four hours after the reperfusion, electron microscopy of the injured tissue showed substantially more platelet aggregation and occlusive platelet microthrombi in the capillaries of the damaged areas of the wild type mice than in those of the FcR gamma(-/-) mice. Platelet Syk was scarcely activated in the FcR gamma(-/-) mice after myocardial ischemia and reperfusion, but significantly activated in the FcR gamma(+/+) mice. CD11b expression on neutrophils was elevated after myocardial ischemia and reperfusion in both mouse groups, whereas myeloperoxidase activity in the injured areas was significantly lower in the FcRgamma(-/-) mice than in the FcRgamma(+/+) mice. These results suggest that the collagen-induced activation of platelets through the FcR gamma plays a pivotal role in the extension of myocardial ischemia-reperfusion injury. FcRgamma and GPVI may be important therapeutic targets for myocardial ischemia-reperfusion injury. PMID- 16246363 TI - A small protein unique to bacteria organizes rRNA tertiary structure over an extensive region of the 50 S ribosomal subunit. AB - A number of small, basic proteins penetrate into the structure of the large subunit of the ribosome. While these proteins presumably aid in the folding of the rRNA, the extent of their contribution to the stability or function of the ribosome is unknown. One of these small, basic proteins is L36, which is highly conserved in Bacteria, but is not present in Archaea or Eucarya. Comparison of ribosome crystal structures shows that the space occupied by L36 in a bacterial ribosome is empty in an archaeal ribosome. To ask what L36 contributes to ribosome stability and function, we have constructed an Escherichia coli strain lacking ribosomal protein L36; cell growth is slowed by 40-50% between 30 degrees C and 42 degrees C. Ribosomes from this deletion strain sediment normally and have a full complement of proteins, other than L36. Chemical protection experiments comparing rRNA from wild-type and L36-deficient ribosomes show the expected increase in reagent accessibility in the immediate vicinity of the L36 binding site, but suggest that a cooperative network of rRNA tertiary interactions has been disrupted along a path extending 60 A deep into the ribosome. These data argue that L36 plays a significant role in organizing 23 S rRNA structure. Perhaps the Archaea and Eucarya have compensated for their lack of L36 by maintaining more stable rRNA tertiary contacts or by adopting alternative protein-RNA interactions elsewhere in the ribosome. PMID- 16246364 TI - Analysis of conformational changes in 16 S rRNA during the course of 30 S subunit assembly. AB - Ribosome biogenesis involves an integrated series of binding events coupled with conformational changes that ultimately result in the formation of a functional macromolecular complex. In vitro, Escherichia coli 30 S subunit assembly occurs in a cooperative manner with the ordered addition of 20 ribosomal proteins (r proteins) with 16 S rRNA. The assembly pathway for 30 S subunits has been dissected in vitro into three steps, where specific r-proteins associate with 16 S rRNA early in 30 S subunit assembly, followed by a mid-assembly conformational rearrangement of the complex that then enables the remaining r-proteins to associate in the final step. Although the three steps of 30 S subunit assembly have been known for some time, few details have been elucidated about changes that occur as a result of these three specific stages. Here, we present a detailed analysis of the concerted early and late stages of small ribosomal subunit assembly. Conformational changes, roles for base-pairing and r-proteins at specific stages of assembly, and a polar nature to the assembly process have been revealed. This work has allowed a more comprehensive and global view of E.coli 30 S ribosomal subunit assembly to be obtained. PMID- 16246365 TI - Structure, dynamics, and membrane topology of stannin: a mediator of neuronal cell apoptosis induced by trimethyltin chloride. AB - Organotin compounds or alkyltins are ubiquitous environmental toxins that have been implicated in cellular death. Unlike other xenobiotic compounds, such as organomercurials and organoleads, alkyltins activate apoptotic cascades at low concentrations. Trimethyltin (TMT) chloride is amongst the most toxic organotin compounds, and is known to selectively inflict injury to specific regions of the brain. Stannin (SNN), an 88-residue mitochondrial membrane protein, has been identified as the specific marker for neuronal cell apoptosis induced by TMT intoxication. This high specificity of TMT makes SNN an ideal model system for understanding the mechanism of organotin neurotoxicity at a molecular level. Here, we report the three-dimensional structure and dynamics of SNN in detergent micelles, and its topological orientation in lipid bilayers as determined by solution and solid-state NMR spectroscopy. We found that SNN is a monotopic membrane protein composed of three domains: a single transmembrane helix (residues 10-33) that transverses the lipid bilayer at approximately a 20 degrees angle with respect to the membrane normal; a 28 residue unstructured linker, which includes a conserved CXC metal-binding motif and a putative 14-3-3zeta binding domain; and a distorted cytoplasmic helix (residues 61-79) that is partially absorbed into the plane of the lipid bilayer with a tilt angle of approximately 80 degrees from the membrane normal. The structure and architecture of SNN within the lipid environment provides insight about how this protein transmits toxic insults caused by TMT across the membrane. PMID- 16246366 TI - Versatile peroxidase oxidation of high redox potential aromatic compounds: site directed mutagenesis, spectroscopic and crystallographic investigation of three long-range electron transfer pathways. AB - Versatile peroxidases (VP), a recently described family of ligninolytic peroxidases, show a hybrid molecular architecture combining different oxidation sites connected to the heme cofactor. High-resolution crystal structures as well as homology models of VP isoenzymes from the fungus Pleurotus eryngii revealed three possibilities for long-range electron transfer for the oxidation of high redox potential aromatic compounds. The possible pathways would start either at Trp164 or His232 of isoenzyme VPL, and at His82 or Trp170 of isoenzyme VPS1. These residues are exposed, and less than 11 A apart from the heme. With the purpose of investigating their functionality, two single mutations (W164S and H232F) and one double mutation (W164S/P76H) were introduced in VPL that: (i) removed the two pathways in this isoenzyme; and (ii) incorporated the absent putative pathway. Analysis of the variants showed that Trp164 is required for oxidation of two high redox potential model substrates (veratryl alcohol and Reactive Black 5), whereas the two other pathways (starting at His232 and His82) are not involved in long-range electron transfer (LRET). None of the mutations affected Mn2+ oxidation, which would take place at the opposite side of the enzyme. Substitution of Trp164 by His also resulted in an inactive variant, indicating that an indole side-chain is required for activity. It is proposed that substrate oxidation occurs via a protein-based radical. For the first time in a ligninolytic peroxidase such an intermediate species could be detected by low-temperature electron paramagnetic resonance of H2O2-activated VP, and was found to exist at Trp164 as a neutral radical. The H2O2-activated VP was self reduced in the absence of reducing substrates. Trp164 is also involved in this reaction, which in the W164S variant was blocked at the level of compound II. When analyzing VP crystal structures close to atomic resolution, no hydroxylation of the Trp164 Cbeta atom was observed (even after addition of several equivalents of H2O2). This is in contrast to lignin peroxidase Trp171. Analysis of the crystal structures of both peroxidases showed differences in the environment of the protein radical-forming residue that could affect its reactivity. These variations would also explain differences found for the oxidation of some high redox potential aromatic substrates. PMID- 16246367 TI - Conformational changes of Escherichia coli sigma54-RNA-polymerase upon closed promoter complex formation. AB - RNA polymerase from the mesophile Escherichia coli exists in two forms, the core enzyme and the holoenzyme. Using cryo-electron microscopy and single-particle analysis, we have obtained the structure of the complete RNA polymerase from E.coli containing the sigma54 factor within the closed-promoter complex. Comparisons with earlier reconstructions of the core enzyme and the sigma54 holoenzyme reveal the behaviour of this major variant RNA polymerase in defined functional states. The binding of DNA leads to significant conformational changes in the enzyme's catalytic subunits, apparently a necessity for the initiation of enhancer-dependent promoter-specific transcription. PMID- 16246368 TI - Structures of Mycobacterium tuberculosis DosR and DosR-DNA complex involved in gene activation during adaptation to hypoxic latency. AB - On encountering low oxygen conditions, DosR activates the transcription of 47 genes, promoting long-term survival of Mycobacterium tuberculosis in a non replicating state. Here, we report the crystal structures of the DosR C-terminal domain and its complex with a consensus DNA sequence of the hypoxia-induced gene promoter. The DosR C-terminal domain contains four alpha-helices and forms tetramers consisting of two dimers with non-intersecting dyads. In the DNA-bound structure, each DosR C-terminal domain in a dimer places its DNA-binding helix deep into the major groove, causing two bends in the DNA. DosR makes numerous protein-DNA base contacts using only three amino acid residues per subunit: Lys179, Lys182, and Asn183. The DosR tetramer is unique among response regulators with known structures. PMID- 16246369 TI - Fine structure analysis of a protein folding transition state; distinguishing between hydrophobic stabilization and specific packing. AB - Developing a detailed understanding of the structure and energetics of protein folding transition states is a key step in describing the folding process. The phi-value analysis approach allows the energetic contribution of side-chains to be mapped out by comparing wild-type with individual mutants where conservative changes are introduced. Studies where multiple substitutions are made at individual sites are much rarer but are potentially very useful for understanding the contribution of each element of a side-chain to transition state formation, and for distinguishing the relative importance of specific packing versus hydrophobic interactions. We have made a series of conservative mutations at multiple buried sites in the N-terminal domain of L9 in order to assess the relative importance of specific side-chain packing versus less specific hydrophobic stabilization of the transition state. A total of 28 variants were prepared using both naturally occurring and non-naturally occurring amino acids at six sites. Analysis of the mutants by NMR and CD showed no perturbation of the structure. There is no correlation between changes in hydrophobicity and changes in stability. In contrast, there is excellent linear correlation between the hydrophobicity of a side-chain and the log of the folding rate, ln(k(f)). The correlation between ln(k(f)) and the change in hydrophobicity holds even for substitutions that change the shape and/or size of a side-chain significantly. For most sites, the correlation with the logarithm of the unfolding rate, ln(k(u)), is much worse. Mutants with more hydrophobic amino acid substitutions fold faster, and those with less hydrophobic amino acid substitutions fold slower. The results show that hydrophobic interactions amongst core residues are an important driving force for forming the transition state, and are more important than specific tight packing interactions. Finally, a number of substitutions lead to negative phi-values and the origin of these effects are described. PMID- 16246370 TI - Study of the active site residues of a glycoside hydrolase family 8 xylanase. AB - Site-directed mutagenesis and a comparative characterisation of the kinetic parameters, pH dependency of activity and thermal stability of mutant and wild type enzymes have been used in association with crystallographic analysis to delineate the functions of several active site residues in a novel glycoside hydrolase family 8 xylanase. Each of the residues investigated plays an essential role in this enzyme: E78 as the general acid, D281 as the general base and in orientating the nucleophilic water molecule, Y203 in maintaining the position of the nucleophilic water molecule and in structural integrity and D144 in sugar ring distortion and transition state stabilization. Interestingly, although crystal structure analyses and the pH-activity profiles clearly identify the functions of E78 and D281, substitution of these residues with their amide derivatives results in only a 250-fold and 700-fold reduction in their apparent k(cat) values, respectively. This, in addition to the observation that the proposed general base is not conserved in all glycoside hydrolase family 8 enzymes, indicates that the mechanistic architecture in this family of inverting enzymes is more complex than is conventionally believed and points to a diversity in the identity of the mechanistically important residues as well as in the arrangement of the intricate microenvironment of the active site among members of this family. PMID- 16246371 TI - Crystal structure of truncated Fibrobacter succinogenes 1,3-1,4-beta-D-glucanase in complex with beta-1,3-1,4-cellotriose. AB - Fibrobacter succinogenes 1,3-1,4-beta-D-glucanase (Fsbeta-glucanase) catalyzes the specific hydrolysis of beta-1,4 glycosidic bonds adjacent to beta-1,3 linkages in beta-D-glucans or lichenan. This is the first report to elucidate the crystal structure of a truncated Fsbeta-glucanase (TFsbeta-glucanase) in complex with beta-1,3-1,4-cellotriose, a major product of the enzyme reaction. The crystal structures, at a resolution of 2.3 angstroms, reveal that the overall fold of TFsbeta-glucanase remains virtually unchanged upon sugar binding. The enzyme accommodates five glucose residues, forming a concave active cleft. The beta-1,3-1,4-cellotriose with subsites -3 to -1 bound to the active cleft of TFsbeta-glucanase with its reducing end subsite -1 close to the key catalytic residues Glu56 and Glu60. All three subsites of the beta-1,3-1,4-cellotriose adopted a relaxed C(1)4 conformation, with a beta-1,3 glycosidic linkage between subsites -2 and -1, and a beta-1,4 glycosidic linkage between subsites -3 and -2. On the basis of the enzyme-product complex structure observed in this study, a catalytic mechanism and substrate binding conformation of the active site of TFsbeta-glucanase is proposed. PMID- 16246372 TI - Stability of the replicator equation for a single species with a multi dimensional continuous trait space. AB - The replicator equation model for the evolution of individual behaviors in a single species with a multi-dimensional continuous trait space is developed as a dynamics on the set of probability measures. Stability of monomorphisms in this model using the weak topology is compared to more traditional methods of adaptive dynamics. For quadratic fitness functions and initial normal trait distributions, it is shown that the multi-dimensional continuously stable strategy (CSS) of adaptive dynamics is often relevant for predicting stability of the measure theoretic model but may be too strong in general. For general fitness functions and trait distributions, the CSS is related to dominance solvability which can be used to characterize local stability for a large class of trait distributions that have no gaps in their supports whereas the stronger neighborhood invader strategy (NIS) concept is needed if the supports are arbitrary. PMID- 16246373 TI - Violent men have more sons: further evidence for the generalized Trivers-Willard hypothesis (gTWH). AB - The generalized Trivers-Willard hypothesis (gTWH) [Kanazawa, S., 2005a. Big and tall parents have more sons; further generalizations of the Trivers-Willard hypothesis. J. Theor. Biol. 235, 583-590] proposes that parents who possess any heritable trait which increases the male reproductive success at a greater rate than female reproductive success in a given environment have a higher-than expected offspring sex ratio, and parents who possess any heritable trait which increases the female reproductive success at a greater rate than male reproductive success in a given environment have a lower-than-expected offspring sex ratio. One heritable trait which increases the reproductive success of sons significantly more than that of daughters in the ancestral environment is the tendency toward violence and aggression. I therefore predict that violent parents have a higher-than-expected offspring sex ratio (more sons). The analysis of both American samples and a British sample demonstrates that battered women, who are mated to violent men, have significantly more sons than daughters. PMID- 16246374 TI - Renal ischemia/reperfusion injury inhibits differentiation of dendritic cells derived from bone marrow monocytes in rats. AB - Dendritic cells (DCs) are impacted by surgical injury, exercise, and other physiological stressors. This study aims to determine whether renal I/R injury affects 1) the differentiation of myeloid DCs from bone marrow monocytes (BMMos) and the maturation and activation state of these DCs and 2) DC infiltration of kidney. Sprague-Dawley rats were subjected to I/R injury or sham-operated. Creatinine clearance was monitored daily during the 14 d of reperfusion that followed the ischemic insult. At 2 and 14 d of reperfusion, the following were assessed 1) properties of BMMo-derived DCs (i.e., the amount of generated DCs, differentiation state markers [CD11c, CD80, CD86, and Ia], and functional state [MLR and amount of IL-12 produced]), and 2) the presence of DCs in the kidney. Numbers of BMMo-derived DCs were significantly decreased in the I/R injured group (compared with the sham-operated group) at 2 d but not 14 d. A comparison of the their functionality found mixed lymphocyte response [MLR] and IL-12 production were similar in the two groups at both time points. Also, immunohistochemistry showed infiltrating DCs in the outer medulla of the I/R injured kidney at 2 d but not 14 d of reperfusion. Thus, I/R stress reduces the number of DCs differentiated from BMMos but not the functional activity of these DCs. This decrease may reflect a stress-induced downshift in the capacity of BMMos to differentiate into DCs and a parallel upshift in the capacity of DCs to infiltrate the kidney. PMID- 16246375 TI - Effect of ecdysterone on glucose metabolism in vitro. AB - The aims of this study was to investigate whether ecdysterone is able to exert glucose-lowering effect on hepatocytes or stimulate the secretion of insulin. HepG2 cell line was used for glucose consumption (GC) studies. At moderate high glucose concentration (11.1 mmol/L), GC of HepG2 cells was increased by 44% to 77% with ecdysterone 1 x 10(-6) to 1 x 10(-4) mol/L, which was comparable to that with 1 x 10(-3) mol/L metformin. The glucose-lowering effect of ecdysterone decreased as the glucose concentration of medium increased. The maximal potency was reached in the presence of 5.5 mmol/L glucose, and the effect was disappeared as the glucose consumption was increased to 22.2 mmol/L. This effect was independent on insulin concentration, which was similar to that of metformin and was different from that of troglitazone, whose glucose-lowering effect was insulin-dependent. Troglitazone had a better antihyperglycemic potency than metformin when insulin was added. Simultaneously, a significant toxicity of troglitazone to HepG2 cells was observed. betaTC3 cells were not stimulated by ecdysterone, that is, no secretogogue effect of ecdysterone was observed. The results indicate that ecdysterone is able to exert the glucose-lowering effect in hepatocytes which is insulin-independent, but has no effect on insulin release. PMID- 16246376 TI - Relationship between oxidative stress and cognitive impairment in the elderly of rural vs. urban communities. AB - Psychological stress and environmental pollution are frequently associated to urban environment and oxidative stress (OxS). Likewise, OxS is a risk factor for cognitive impairment (CI) in the elderly. Therefore, we hypothesized that the prevalence of CI in subjects of the urban area could be higher than in those of the rural area, and linked to higher OxS. The aim of the study was to determine the relationship between OxS and CI in elderly individuals from rural and urban settings in Mexico. Plasmatic TBARS, plasma total antioxidant status, and the antioxidant enzymes superoxide dismutase (SOD) and glutathione peroxidase (GPx) were measured in 104 urban and 85 rural elderly individuals. Cognitive functions were evaluated through the Mini Mental State Examination. We found a greater proportion of subjects with OxS and CI in urban than in rural areas (25% vs. 9%), with an odds ratio of 5.67 (CI95% 1.14-38.02, p < 0.05). Our findings suggest that the elderly in urban areas have more OxS and a higher risk of developing CI compared with elderly individuals in a rural environment. PMID- 16246378 TI - Modelling salmon lice dispersal in Loch Torridon, Scotland. AB - A particle transport model is described that is being used to simulate the dispersal of salmon lice (Lepeophtheirus salmonis) larvae in the waters of Loch Torridon. A hydrodynamic model, forced by tides and winds, drives the transport model. Particle movements are strongly influenced by winds, which can lead to formation of lice concentrations in coastal areas several kilometres from the source. Idealised constant wind simulations have been used to locate areas that larval lice may potentially reach from given source locations. Detailed analysis of simulations forced with real wind data is required to assess areas that larval lice from these sources are likely to reach. Further field and experimental work on the viability of lice is required to assess infection risk. PMID- 16246377 TI - PCDD and PCDF intake through consumption of locally produced seafood by Venice lagoon residents: elements for risk management. PMID- 16246379 TI - The impact of sika deer grazing on the vegetation and infauna of Arne saltmarsh. AB - Arne saltmarsh, an RSPB reserve, is situated in Poole Harbour on the English south coast. In recent years, there has been concern about possible changes in the suitability of the site for Redshank (Trigna totanus) due to sika deer (Cervus nippon) grazing. In order to assess these changes, 50 plots were established in three different locations: 20 in grazed areas, 20 in ungrazed areas and 10 fenced enclosures. Deer grazing was found to significantly affect structural and species diversity of the saltmarsh vegetation. Spartina anglica dominated in ungrazed areas whilst Salicornia ramosissima and, to a lesser extent, Puccinellia maritima dominated in grazed sites. In grazed areas the vegetation cover was significantly lower, as was vegetation height and volume. In addition, significant changes were observed in the root biomass, which was lower in grazed areas. Infaunal diversity was generally low throughout the survey area. However, significant variations were observed. Invertebrates abundance was more abundant in grazed plots than in ungrazed plots, and least abundant in fenced plots. The study indicated that in its current condition, localised areas of Arne saltmarsh do not provide adequate habitat requirements for Tringa totanus. PMID- 16246380 TI - The impact of a macroalgal mat on benthic biodiversity in Poole Harbour. AB - Blooms of macroalgal matting are increasingly common within temperate zones and are often comprised of opportunistic species such as Ulva lactuca. Where this algae forms a dense mat, a stressful environment is created in the sediment below, influencing the invertebrate infaunal assemblage. This study was conducted over a six month period during which a dense mat of U. lactuca developed and subsequently dispersed. The algal mat was found to have a significant negative impact on species richness, abundance and biomass of the macroinfauna. However, a faunal community developed within the algal mat which contained several species not previously observed. This community increased the abundance and diversity of the overall invertebrate assemblage. The results are discussed in relation to impacts on the ecosystem as a whole. PMID- 16246381 TI - Environmental influences on food choice, physical activity and energy balance. AB - In this paper, the environment is defined as the macro- and community-level factors, including physical, legal and policy factors, that influence household and individual decisions. Thus, environment is conceived as the external context in which household and individual decisions are made. This paper reviews the literature on the ways the environment affects diet, physical activity, and obesity. Other key environmental factors discussed include economic, legal, and policy factors. Behind the major changes in diet and physical activity in the US and globally lie large shifts in food production, processing, and distribution systems as well as food shopping and eating options, resulting in the increase in availability of energy-dense foods. Similarly, the ways we move at home, work, leisure, and travel have shifted markedly, resulting in substantial reductions in energy expenditure. Many small area studies have linked environmental shifts with diet and activity changes. This paper begins with a review of environmental influences on diet and physical activity, and includes the discussion of two case studies on environmental influences on physical activity in a nationally representative sample of US adolescents. The case studies illustrate the important role of physical activity resources and the inequitable distribution of such activity-related facilities and resources, with high minority, low educated populations at strong disadvantage. Further, the research shows a significant association of such facilities with individual-level health behavior. The inequity in environmental supports for physical activity may underlie health disparities in the US population. PMID- 16246382 TI - Birch PR-10c interacts with several biologically important ligands. AB - PR-10c is a unique member of PR-10 proteins in birch, since it is the only one known to be post-translationally modified by glutathione and is not constitutively expressed in pollen. Both reduced and S-glutathiolated forms of PR 10c show low ribonuclease activity. However, the major function of the protein is apparently not yet resolved. Our protein-ligand interaction studies with saturation transfer difference (STD) NMR revealed that PR-10c interacts with several biologically important molecules, including cytokinin, flavonoid glycosides, sterols and emodin. Competition study with deoxycholate and kinetin revealed no statistically significant binding interference, indicating that these ligands have different binding sites in PR-10c. Ligand docking studies with a molecular model of PR-10c support the STD NMR results of ligand binding and binding epitopes, suggesting that there are three potential binding sites in PR 10c: two in the hydrophobic cavity and one in the glycine-rich loop. Our docking calculations suggested that only kinetin interacts with the glycine-rich loop, the binding occurring through its adenine moiety. Clear ligand specificity could be observed in the binding of nucleotide derivatives. S-glutathiolation of PR-10c did not affect kinetin binding. The present results suggest that birch PR-10c is a multifunctional protein, which has diverse roles in plant stress responses. PMID- 16246383 TI - Fluoroquinolone-resistant Vibrio cholerae isolated during a cholera outbreak in India. AB - During the cholera epidemic of 2002 in and around Hubli, south India, Vibrio cholerae strains resistant to fluoroquinolones were isolated. Among the isolates of V. cholerae non-O1, non-O139 serogroups, 55.9% and 47.1% were resistant to norfloxacin and ciprofloxacin, respectively. However, only 12.5% of the O1 serogroup strains were resistant to both norfloxacin and ciprofloxacin. Though the O139 serogroup strains were susceptible to these antibiotics, they exhibited multidrug resistance. Emergence of fluoroquinolone-resistant V. cholerae that also exhibited multidrug resistance is of great significance in the epidemiology and control of cholera. PMID- 16246384 TI - Using remote sensing and geographic information systems to identify villages at high risk for rhodesiense sleeping sickness in Uganda. AB - Geographic information systems (GIS) and remote sensing were used to identify villages at high risk for sleeping sickness, as defined by reported incidence. Landsat Enhanced Thematic Mapper (ETM) satellite data were classified to obtain a map of land cover, and the Normalised Difference Vegetation Index (NDVI) and Landsat band 5 were derived as unclassified measures of vegetation density and soil moisture, respectively. GIS functions were used to determine the areas of land cover types and mean NDVI and band 5 values within 1.5 km radii of 389 villages where sleeping sickness incidence had been estimated. Analysis using backward binary logistic regression found proximity to swampland and low population density to be predictive of reported sleeping sickness presence, with distance to the sleeping sickness hospital as an important confounding variable. These findings demonstrate the potential of remote sensing and GIS to characterize village-level risk of sleeping sickness in endemic regions. PMID- 16246385 TI - Synthesis of 3alpha-hydroxy-21-(1'-imidazolyl)-3beta-methoxyl-methyl-5alpha pregnan-20-one via lithium imidazole with 17alpha-acetylbromopregnanone. AB - The synthesis of biologically active 3alpha-hydroxyl-21-(1'-imidazolyl)-3beta methoxymethyl-5alpha-pregnan-20-one was accomplished in six steps. The key steps were the improvement of stereoselectivity for acetyl isomers in C-17 and the introduction of imidazole into the core structure by use of lithium imidazole. This latter key step provided the desired product in 82% yield without the formation of 1,3-disubstituted imidazolium salt as impurity, which is generally observed in traditional method. PMID- 16246386 TI - Asymptotically exact analysis of stochastic metapopulation dynamics with explicit spatial structure. AB - We describe a mathematically exact method for the analysis of spatially structured Markov processes. The method is based on a systematic perturbation expansion around the deterministic, non-spatial mean-field theory, using the theory of distributions to account for space and the underlying stochastic differential equations to account for stochasticity. As an example, we consider a spatial version of the Levins metapopulation model, in which the habitat patches are distributed in the d-dimensional landscape Rd in a random (but possibly correlated) manner. Assuming that the dispersal kernel is characterized by a length scale L, we examine how the behavior of the metapopulation deviates from the mean-field model for a finite but large L. For example, we show that the equilibrium fraction of occupied patches is given by p(0)+c/L(d)+O(L(-3d/2)), where p(0) is the equilibrium state of the Levins model and the constant c depends on p(0), the dispersal kernel, and the structure of the landscape. We show that patch occupancy can be increased or decreased by spatial structure, but is always decreased by stochasticity. Comparison with simulations show that the analytical results are not only asymptotically exact (as L-->infinity), but a good approximation also when L is relatively small. PMID- 16246387 TI - Conversion of Trypanosoma cruzi Tc52 released factor to a protein inducing apoptosis. AB - In this study Tc52, a Trypanosoma cruzi released protein, which exerts an immunoregulatory activity, was converted to a molecular form with altered biological function. Indeed, the genetic fusion of Tc52 to a carrier protein, the Shistosoma japonicum glutathione S-transferase (Tc52-Sj26), was shown to induce apoptosis in spleen cells from BALB/c or CBA mice and the human T-cell leukemic cell line (CEM). Cell death by apoptosis was evidenced by the following criteria: (1) increased binding of Annexin V to rTc52-treated spleen cells; (2) the presence of an ordered cleavage of the DNA backbone; (3) double labeling showed increased number of T cells undergoing apoptosis upon incubation with rTc52; (4) the use of a CEM cell line and TUNEL assay allowed to show in situ DNA fragmentation. Surprisingly, intraperitoneal injections of rTc52 to BALB/c mice, which were then infected with T. cruzi, resulted in increased parasiteamia levels and is congruent to 2.5 times increase of macrophages number. Since native Tc52 could not trigger, apoptosis of T cells we could hypothesized that the fusion of Tc52 with Sj26 led to conformational changes resulting in apoptosis inducing properties of rTc52. The possible in vivo physiopathological implications of these finding were discussed. PMID- 16246388 TI - Changes in the carbon dioxide expirogram in response to ozone exposure. AB - The objectives of this study were to quantify pulmonary responses to ozone (O3) exposure by parameters computed from the carbon dioxide expirogram and to compare these responses to decrements in forced expired spirometry. Anatomical dead space (VD) was determined from the pure dead space and transition regions of the expirogram. Four alternative parameters were computed from the alveolar plateau: slope (S), normalized slope (NS), peripheral cross-sectional area (AP) and well mixed peripheral volume (VMP). Forty-seven healthy nonsmokers (25 men and 22 women) participated in two research sessions in which they exercised on a cycle ergometer for 1 h while orally inhaling either room air at a minute ventilation of 30.6 +/- 3.6 L or room air mixed with 0.252 +/- 0.029 ppm O3 at a minute ventilation of 29.9 +/- 3.7 L. Carbon dioxide expirograms were measured before exposure, 10 min after exposure and 70 min after exposure. Percent changes (mean +/- SD) in expirogram parameters were significant (P < or = 0.002) at both 10 and 70 min after O3 exposure: VD(-4.2 +/- 5.1, -3.3 +/- 6.9), S(16.4 +/- 17.9, +15.1 +/- 20.2), NS(17.5 +/- 15.4, +15.9 +/- 19.2), AP(-8.1 +/- 7.6, -7.7 +/- 9.8) and VMP(-15.4 +/- 13.0, -13.0 +/- 15.2). Percent decrements of forced expired volume in one second (FEV1) were also significant at both 10 min (-13.3 +/- 13.4) and 70 min (-11.1 +/- 9.2) following O3 exposure. Changes in the expirogram as well as decrements in FEV1 were not significant at either time point after air exposure. Thus, the CO2 expirogram is useful for characterizing the effect of O3 exposure on gas transport, and for supplementing forced expired spirometry that is frequently used to quantify lung mechanics. PMID- 16246389 TI - Depth gauging of defects using low frequency wideband Rayleigh waves. AB - In non-destructive testing for cracks it is not sufficient to simply detect the presence of a defect, but it is essential to have an accurate measure of the depth. Accurate calibration of the techniques used to gauge defect size is therefore necessary. Recent progress in the field of non-contact ultrasonic testing has led to the development of a practically viable system for generating and detecting wideband Rayleigh waves on electrically conducting or magnetic samples using electro-magnetic acoustic transducers (EMATs). This system has been used to gauge the depth and position of surface breaking defects, and has many applications including metal billet testing and detecting and sizing gauge corner cracking in rails. In this paper we report experiments calibrating the response of EMATs when a defect is present between the generator and receiver, using a calibration sample with slots machined perpendicular to the surface to simulate surface breaking cracks. The depth of the defect can be gauged in the time domain and frequency domain, with an accurate 'fingerprint' of the position given by an enhancement of the signal when the receiver is close to the defect. The best choice of EMAT design for different applications is discussed, as is the best position for the receive EMAT to avoid areas of interference between the Rayleigh wave and bulk waves diffracted from the crack tip. PMID- 16246390 TI - Human parvovirus B19 infection of monocytic cell line U937 and antibody-dependent enhancement. AB - Human parvovirus B19 (B19) infects human erythroid lineage cells. Accumulating evidence also shows that B19 is detectable in nonerythroid lineage cells in vivo, but the mechanism of infection is still not clear. In this study, we explored the mode of B19 infection of human monocytic cell line U937. An in vitro infection study demonstrated B19 binding of U937 and slow replication of B19-DNA with B19 NS1 mRNA transcription. B19-DNA replication in U937 was accompanied by undetectable level of B19-VP1 mRNA transcription, indicating that B19 infection of U937 cells may be abortive. Levels of B19-DNA and B19-NS1 mRNA transcription increased in the presence of anti-B19 IgG antibodies, but this effect decreased in the presence of anti-Fc receptor antibodies, showing antibody-dependent enhancement by B19 infection. Antibody-dependent enhancement also caused the increased production of TNFalpha in U937. This study is the first to suggest B19 infection of nonerythroid lineage cells with antibody-dependent enhancement. PMID- 16246391 TI - CTL escape and increased viremia irrespective of HIV-specific CD4+ T-helper responses in two HIV-infected individuals. AB - We investigated whether development of mutations leads to loss of CD8 T-cell recognition in HIV-1 infection and is possibly linked to alterations in HIV-1 specific CD4(+) T-cell responses in 2 HIV-infected individuals. In patient, H434 full genome sequencing of HIV-1 biological clones at early and late time points during disease progression showed development of fixed mutations in 16 predicted HIV-specific CTL epitopes. Loss of T-cell recognition and reactivity against wild type and mutant epitopes was observed primarily for the HLA-B27-restricted KK10 epitope and HLA-A2-restricted SL9 epitope. Similarly, in patient H671, decreasing numbers of HLA-A3-restricted CD8(+) T cells specific for the wild-type RK9 epitope was observed after CTL escape. Only in patient H434 loss of CTL responses was paralleled by a decrease in HIV-specific IL-2(+) CD4(+) T-helper responses. This suggests that loss of T-cell reactivity may not be directly linked to HIV specific CD4(+) T-cell responses but that increased viremia after CTL escape may influence CD4(+) T-helper responses. PMID- 16246392 TI - Matching complementing functions of transformed cells with stable expression of selected viral genes for production of E1-deleted adenovirus vectors. AB - Production of E1-deleted adenovirus (rAd) vectors requires complementation by E1A and E1B functions provided by the production cell line. The two cell lines most commonly used for production of rAd vectors, 293 and Per.C6, were derived from human primary cells and contain contiguous E1A and E1B sequences from the Ad genome. As an alternative system, we tested complementation of rAd vectors using sequential transfection of individual E1A and E1B expression cassettes into A549 human lung tumor cells, which support highly efficient replication of wild type adenovirus. We found that E1A function could be complemented in A549 cells by the mutant E1Adl01/07, and that E1B function could be provided in such cells using only the 55K E1B gene. Production yields in the resulting producer cell line, designated SL0003, were similar to those obtained from 293 cells without generation of detectable recombinant replication competent adenovirus. PMID- 16246393 TI - The effect of binocular and monocular distractors on saccades in participants with normal binocular vision. AB - We tested the effect of visual distractors presented monocularly and binocularly on saccade latency and accuracy to determine whether differences occur in saccadic planning with binocular or monocular visual input. For five participants with normal binocular single vision (BSV), saccade latency and accuracy were compared with distractors presented to the dominant eye, non-dominant eye or to both eyes. Eye movements of the dominant eye were recorded using a Skalar infra red recorder. In the presence of normal BSV, the effect of distractors is significantly larger for saccade latency and accuracy with binocular distractor presentation than for monocular presentations, with no difference between distrators presented to the dominant or non-dominant eye. The implications of these results are discussed with regard to saccade programming. PMID- 16246394 TI - Removing low ppb level perchlorate, RDX, and HMX from groundwater with cetyltrimethylammonium chloride (CTAC) pre-loaded activated carbon. AB - Perchlorate contaminates vast amounts of groundwater throughout the United States which could potentially be used as potable water. Activated carbon pre-loaded with cetyltrimethylammonium chloride has been shown in this research to be an effective adsorbent for removing perchlorate from three low conductivity (50-66 microS/cm) groundwaters containing perchlorate (ClO(4)(-)) concentrations of 0.85, 1.0, and 5.6 parts per billion (ppb), respectively. In rapid small-scale column tests (RSSCTs), the virgin granular activated carbon (GAC) (used as a control) treated between 20,000 and 40,000 bed volumes (BV) of water. In contrast, the activated carbon that was pre-loaded with CTAC processed 170,000 270,000 BV before perchlorate was detected above 0.25 ppb in the effluent. Though this pre-loading significantly increased the capacity for perchlorate, it also diminished the GAC's capacity to remove organics. The groundwater containing 1 ppb ClO(4)(-) also contained the nitro-organics HMX (0.6 ppb) and RDX (5.5-6.6 ppb). RDX was detected in the effluent from the CTAC-pre-loaded bed after only 8000 BV had been processed whereas 308,000 BV could be processed through the virgin bed before RDX was detected. Likewise, HMX breakthrough was observed after 116,000 BV in the CTAC-pre-loaded bed while the virgin RSSCT exhibited no breakthrough of HMX during a test that was operated for 309,000 BV. However, by combining a CTAC-pre-loaded bed followed by a virgin GAC bed in series, both perchlorate and RDX could be removed for the same length of time. PMID- 16246395 TI - Sulphate reduction and the removal of carbon and ammonia in a laboratory-scale constructed wetland. AB - Sulphate is a normal constituent of domestic wastewater and reduced sulphur compounds are known to be potent inhibitors of plant growth and certain microbial activities. However, the knowledge about sulphate reduction and the effect on the removal of C and N in constructed wetlands is still limited. Investigations in laboratory-scale constructed wetland reactors were performed to evaluate the interrelation of carbon and nitrogen removal with the sulphate reduction by use of artificial domestic wastewater. Carbon removal was found to be only slightly affected and remained at high levels of efficiency (75-90%). Only at sulphate reduction intensities above 75 mgl(-1) (50% removal), a decrease of carbon removal of up to 20% was observed. A highly contrary behaviour of ammonia removal was found in general, which decreased exponentially from 75% to 35% related to a linear increase of sulphate reduction up to 75 mgl(-1) (50% removal). Since sulphate removal is considered to be dependant on the load of electron donors, the carbon load of the system was varied. Variation of the load changed the intensities of sulphate reduction immediately, but did not influence the carbon removal effectiveness. Doubling of the carbon concentration of 200 mgl(-1) BOD(5) for domestic wastewater usually led to sulphate reduction of up to 150 mgl(-1) (100% removal). The findings show that, particularly in constructed wetland systems, the sulphur cycle in the rhizosphere is of high importance for performance of the waste water treatment and may initiate a reconsideration of the amount of sulphate present in the tap water systems. PMID- 16246396 TI - Removal of cosmetic ingredients and pharmaceuticals in sewage primary treatment. AB - Two physico-chemical processes, coagulation-flocculation and flotation, have been assessed for enhancing the removal of some selected pharmaceutical and personal care products (PPCPs) present in sewage. Eight compounds, representative of three main groups of PPCPs according to their physico-chemical properties, have been selected: lipophilic compounds (the synthetic musks Galaxolide and Tonalide), neutral compounds (the tranquillizer Diazepam and the antiepileptic Carbamazepine) and acidic compounds (the anti-inflammatories Ibuprofen, Naproxen and Diclofenac). During the coagulation-flocculation assays, the main parameters considered were the selection of the additives, their doses and the temperature of operation (12 or 25 degrees C). Musks-which are highly lipophilic and Diclofenac-with significant sorption affinity-were removed around 50-70% at both temperatures independently of the dose and type of coagulant used. However, the rest of the compounds, which are more hydrophilic, were affected to a lesser degree (with maximum reductions below 25%). The exceptions to this behavior were Carbamazepine and Ibuprofen, which were not removed under any condition tested. During the flotation assays, the parameters studied were the initial content of fat in wastewaters and temperature. Again, musks were removed to a greater degree (35-60%), followed by Diazepam (40-50%) and Diclofenac (20-45%) and, to a lesser extent, Carbamazepine (20-35%), Ibuprofen (10-25%) and Naproxen (10-30%). The best results were always obtained at 25 degrees C, although in some cases the operation at 12 degrees C gave similar results. The removal of musks and neutral compounds was higher in wastewaters with a high fat content (around 150 mgl(-1)). PMID- 16246397 TI - The occurrence and removal of phthalates in a trickle filter STW. AB - This study investigated the fate of phthalates in a trickle filter sewage treatment works. A wide variety of phthalates were researched of which only two were present in significant amounts. Mean concentrations of di-(2-ethylhexyl) phthalate (DEHP) and diethyl phthalate (DEP) measured throughout the system were 23.6 and 25.0 microg/l in raw sewage, 22.0 and 24.8 microg/l in primary, 14.6 and 0.60 microg/l in trickle filter, 18.6 and 0.10 microg/l in humus tank and 18.5 and 0.40 microg/l in reedbed effluents, respectively. Removal by the trickle filter was constantly high for DEP (94-99%) whereas DEHP was variable (<1-44%). Mean concentrations of DEHP and DEP in raw sludge were 30.2 and 1.60 microg/g dry wt, respectively. A mass balance for DEHP was calculated using data from field studies and estimates of sludge production at the works. The mass balance approach helped to provide information that could be used to improve the design and operation of sewage treatment works. PMID- 16246398 TI - Quantitative structure-toxicity relationships of organophosphorous pesticides to fish (Cyprinus carpio). AB - This study was conducted to determine the relationships between 1381 chemical and structural parameters of 43 organophosphorus pesticides (OPs) and their toxicity to fish, Cyprinus carpio, using ChemOffice 8.03 and Dragon 2.1. By multivariate linear regression and intervariable regression analyses, various equations have been derived to calculate the lethal toxicity value, LC(50), for 43 OPs found in fish with different levels of toxicity. Results show that for all selected OPs, especially those of low toxic OPs (LC(50)< 2.5 mM), the equation, LC(50) = 56.259 - 13.071 lg K(ow)+17.510 MATS8P-17.455 Mor24u - 0.085 MW + 1.706 (lg K(ow))(2) + 2.306 (Mor14e)(2) + 6.849 Mor20 m (n = 43, F = 36.815, r = 0.942, r(adj)(2) = 0.862, SE = 2.899, p < 10(-6)), could account for 86.2% of the variability of the toxic effect. The steric and electronic characteristics and the hydrophobicity of OPs, in particular, are among the most important parameters determining the toxicity of OPs to fish. For the OPs with high toxicity, different structural parameters were introduced into the following two equations: LC(50)=3.795-1.195 (H1p)(2)-0.037 U-2.225 MATS3v-19.593 Tcon (n = 16, F = 56.820, r = 0.977, r(adj)(2) = 0.937, SE = 0.143, p < 10(-6)), where LC(50) is less than 2.5 mM, and LC(50) = 0.341-0.561 (HOMA)(2) + 0.231 HOMA (n = 3,r(adj)(2) = 1), where LC(50) is less than 0.3 mM. These results suggest that chemical and structural parameters could be useful in modeling chemical reactivity within homologous series of OP compounds and elucidating possible mechanisms associated with different levels of toxicity to fish. PMID- 16246399 TI - Inguinal sentinel node dissection versus standard inguinal node dissection in patients with vulvar cancer: A comparison of the size of metastasis detected in inguinal lymph nodes. AB - OBJECTIVE: The emergence of sentinel lymph node (SLN) technology has provided the ability for an in depth pathologic evaluation for the detection of metastasis to lymph nodes through the use of ultra-staging. The SLN has been shown to be predictive of the metastatic status of its nodal basin. More recently, SLN dissections have been employed in the evaluation of the inguinal lymphatic basins in patients with vulvar malignancies. We hypothesize that the average size of metastasis detected in non-palpable inguinal lymph nodes is smaller when detected through the use of SLN dissection and ultra-staging versus complete inguinal node dissection (CND). METHODS: This was an IRB approved retrospective study. The tumor registry database was searched to identify all patients diagnosed with a vulvar malignancy from 1990 to 2004. The records were reviewed to identify patients with inguinal lymph node metastasis. Only patients with non-palpable inguinal lymph nodes (metastasis 1 cm or less) were included in the analysis. All pathology slides were reviewed. The smallest metastatic foci of cells were measured from lymph nodes obtained through the traditional complete inguinal lymph node dissection (CND) and compared with the largest metastatic foci of cells detected in sentinel lymph node dissections. The mean size and standard deviation for each group was calculated and analyzed with a Mann-Whitney test. RESULTS: There were 336 inguinal node dissections performed in patients identified with a vulvar malignancy. SLN dissections were performed in 52 groins and CND in 284 groins. Fifty-eight patients were found to have metastatic disease to the inguinal lymph nodes. Thirty of these patients had no evidence of lymph node metastasis on clinical exam or at the time of their EUA. There were 7 groins with metastasis detected through an SLN and 23 groins through a CND. The mean size of the metastatic foci detected in the SLN group was 2.52 mm (SD 1.55) and in the CND group was 4.35 mm (SD 2.63). This was not statistically significant (P = 0.109). However, when comparing the detection of micrometastasis in each set, there was a significant difference (P = 0.02) in the detection of the size of metastasis detected with smaller cluster of cells detected in the SLN group. CONCLUSION: SLN dissection with ultra-staging allows for a more extensive pathologic examination of lymph nodes and may allow for the detection of smaller tumor foci than the traditional pathological examination of lymph nodes obtained from a CND. The clinical implication of the detection of these micrometastasis and smaller metastasis remains to be determined. PMID- 16246400 TI - Placental site trophoblastic tumor: A study of 55 cases and review of the literature emphasizing factors of prognostic significance. AB - OBJECTIVE: The placental site trophoblastic tumor is a rare form of gestational trophoblastic disease. Fifteen percent of reported cases have been fatal, but predicting behavior in individual patients has been challenging. METHODS: The clinical, gross and histopathological features of 55 cases and 180 cases in the literature were analyzed for their effect on survival and in relation to tumor stage. RESULTS: The 55 patients in our series were 20 to 62 (average 32) years of age. The tumors occurred on an average of 34 months after the last known gestation. 84% were stage I, 2% stage II, 5% stage III, and 9% stage IV. Serum levels of human chorionic gonadotropin (hCG) were elevated (average 691 mIU/ml) in 77% of the cases. The tumors were on average 5 cm in greatest dimension and were composed microscopically of infiltrative sheets of intermediate (extravillous) trophoblastic cells. The mitotic rate ranged from 0 to 20 (average 5.0) per 10 high power fields. The follow-up interval averaged 4.6 years. Eight patients (15%) died from metastatic tumor, and nine additional patients had metastases or a recurrence but were alive at last contact. The most common metastatic sites were the lungs, liver, and vagina. CONCLUSIONS: Significant factors associated with adverse survival in the present series were age over 35 years (P = 0.025), interval since the last pregnancy of over 2 years (P = 0.014), deep myometrial invasion (P = 0.006), stage III or IV (P < 0.0005), maximum hCG level > 1000 mIU/ml (P = 0.034), extensive coagulative necrosis (P = 0.024), high mitotic rate (P = 0.005), and the presence of cells with clear cytoplasm (P < 0.0005). Only stage and clear cytoplasm were independent predictors of overall survival, while stage and age were the only independent predictors of time to recurrence or disease-free survival. In the literature, factors associated with survival were stage (P < 0.005), interval from preceding pregnancy of over 2 years (P = 0.029), previous term pregnancy (P = 0.046), high mitotic rate (P < 0.0005), and high hCG level (P = 0.037). PMID- 16246401 TI - Ovarian cancer, diagnosed with PET, with bilateral inguinal lymphadenopathy as primary presenting sign. AB - BACKGROUND: Ovarian cancer groin lymph nodes metastases are rare. Only one case of disease spread isolated to the groins has been reported in the English literature. We report the case of a patient with bilateral inguinal lymphadenopathy due to tumor metastases and in which tumor origin was diagnosed using [(18)F]fluorodeoxyglucose positron emission tomography (PET). CASE REPORT: This is the first case of bilateral groin lymph nodes as sole site of metastases from ovarian cancer. Primary origin was identified using PET. CONCLUSIONS: In rare cases, the ovaries can directly metastasis to the groin even in a relatively initial tumor growth phase. PET may play a role in the diagnosis of occult ovarian tumors. PMID- 16246402 TI - Does size matter? Tumor size and morphology as predictors of nodal status and recurrence in endometrial cancer. AB - OBJECTIVE: To determine whether tumor size or morphology is predictive of extrauterine disease and/or recurrence risk in endometrial cancer and therefore guide decisions about the necessity of complete surgical staging and adjuvant therapy. METHODS: All women with surgically treated endometrial carcinoma between 1 January 1990 and 1 January 2000 were eligible. 345 patients were eligible for retrospective chart review. Univariate and multivariate logistic regression models were used to determine the predictors of nodal metastasis and recurrence. RESULTS: As tumor size increased, the risk of nodal metastasis increased. However, a risk of nodal metastasis remained even with small lesions less than or equal to 2 cm (6.3% risk). Patients with tumor size greater than 2 cm had a 26.3% incidence of nodal metastasis. In univariate analysis, the odds ratio (OR) for tumor size as a predictor of extrauterine disease was 1.4 (95% CI 1.2-1.6). In multivariate analysis, tumor size was not statistically significant. Only the lesions greater than or equal to 8 cm confer a risk that approaches previously identified well-known predictors. Tumor size was not found to be a statistically significant predictor of recurrence OR 1.3 (1.0-1.8). CONCLUSIONS: Tumor size correlates with extrauterine disease, but it is not an independent prognostic variable. Although the risk of extrauterine disease increases with tumor size, the risk of nodal metastases remains even for those patients with very small tumors, underscoring the need for routine complete surgical staging. Tumor size does not appear to be an independent predictor of recurrence. PMID- 16246403 TI - Aggressive angiomyxoma of the vulva: Dramatic response to gonadotropin-releasing hormone agonist therapy. AB - BACKGROUND: Aggressive angiomyxoma (AA) is a vulvovaginal mesenchymal neoplasm with a marked tendency to local recurrence but which usually does not metastasise. Most cases exhibit positive immunohistochemical staining with oestrogen receptor (ER) and, or, progesterone receptor (PR). CASE: We report a case of AA which exhibited positive immunohistochemical staining with ER and in which radiological examination following resection showed extensive residual tumour. The patient was commenced on gonadotropin-releasing hormone (GnRH) agonist therapy which resulted in complete radiological remission with replacement by scar tissue. The patient is currently maintained on a GnRH agonist. CONCLUSION: The present case, together with several others reported in the literature, suggests that GnRH agonists may be of value in managing cases of AA, either primary or recurrent, which are not amenable to surgical excision. These agents may also be used to effect a reduction in size, so that more conservative surgery can be undertaken. PMID- 16246404 TI - HPV16 E2 gene disruption and polymorphisms of E2 and LCR: some significant associations with cervical cancer in Indian women. AB - OBJECTIVES: We evaluated the status of the HPV16 E2 gene (disrupted or intact), nucleotide sequence alterations within intact E2 genes and LCR of HPV16 isolates in a group of CaCx cases (invasive squamous cell carcinomas, n = 81) and population controls (normal cervical scrapes, n = 27) from Indian women. METHODS: E2 disruption was detected by amplifying the entire E2 gene with single set of primers, while overlapping primers were used to determine if any particular region got selectively disrupted. Nucleotide variations in E2 and LCR were analyzed by PCR amplification followed by bi-directional sequencing. The associations between the viral factors and CaCx were analyzed using Fisher's Exact or Chi-squared test and interpreted as OR (95% CI) and P values. RESULTS: E2 disruption was significantly higher among the cases [3.38 (1.07-10.72); P = 0.02], which was maximum in the region between nucleotides 3650 and 3872 (DNA binding region). The European (E) variant was found to be the prevalent subgroup (87.76% among cases and 96.30% among the controls), and the remaining samples were Asian-American variants. Among the E subgroup, variation at position 7450 (T > C) within the E2-binding site-IV was found to be significantly higher among the E2 undisrupted cases (21/37; 56.76%), compared to controls (5/18; 27.78%) [3.41 (1.01-11.55); P = 0.03]. CONCLUSIONS: Besides HPV16 E2 disruption, LCR 7450T > C variation within undisrupted E2 of E subgroup appears to be a major factor contributing to the risk of CaCx development in Indian women. Furthermore, polymorphisms in the E2 gene of HPV16 may not be significant for disease risk. PMID- 16246405 TI - Prevalence and prognostic significance of COX-2 expression in stage IB cervical cancer. AB - OBJECTIVES: To evaluate the prevalence of cyclooxygenase-2 (COX-2), correlation with various clinicopathologic factors and prognostic significance of COX-2 in stage IB cervical cancer patients. METHODS: 89 paraffin-embedded specimens of patients with stage IB cervical cancer underwent radical hysterectomy and pelvic lymphadenectomy at King Chulalongkorn Memorial Hospital during 1 January 1997-31 December 2002 and were stained with polyclonal goat antiserum against COX-2 using immunohistochemical method. Medical records were reviewed; clinicopathological variables were retrieved and used for analysis. RESULTS: The prevalence of positive COX-2 expression in stage IB cervical cancer in this study was 49.4%. Positive COX-2 expression in cervical adenocarcinoma was higher than squamous cell carcinoma (86.7% versus 40.6%, P < 0.05) and significantly expressed when lymph node metastasis was presented (100% versus 46.4%, P < 0.05). However, COX-2 expression was possibly associated with parametrial involvement (80% versus 47.6%, P > 0.05). There was no correlation between COX-2 expression and patient's age, tumor size, depth of stromal invasion and lymphovascular space invasion. Five-year disease free survival and 5-year overall survival in patients with positive COX-2 expression were 81% and 98% which were not differed from patients with negative COX-2 expression (92% and 95%, P > 0.05). CONCLUSIONS: Strong correlation was found in cervical adenocarcinoma and lymph node metastasis. However, COX-2 expression failed to demonstrate as a significant prognostic factor in stage IB cervical cancer. PMID- 16246406 TI - Relation of smoking and drinking to sleep disturbance among Japanese pregnant women. AB - BACKGROUND: Pregnant women suffer from sleep disturbance, which may be aggravated by smoking and/or drinking. We investigate here the joint effect of smoking and drinking with respect to sleep disturbance during pregnancy. METHODS: Survey of about 16,000 pregnant women in Japan, conducted in 2002 using a self-administered questionnaire. RESULTS: Both smoking and drinking increased the odds of sleep disturbances, such as subjective insufficient sleep, difficulty in initiating or maintaining sleep, early-morning awakening, short sleep duration, excessive daytime sleepiness and restless legs syndrome. The joint odds ratios for smoking and drinking corresponded more or less to the products of the odds ratio for smoking or drinking. CONCLUSION: Smoking and drinking are independently associated with increased sleep disturbance during pregnancy, in addition to their other well-known side-effects. PMID- 16246407 TI - Importance of sperm genotype (indicus versus taurus) for fertility and embryonic development at elevated temperatures. AB - Heat stress has negative effects on bovine reproduction, particularly for European breeds (Bos taurus taurus) that are less thermotolerant than zebu cattle (Bos taurus indicus). Here, the evidence that spermatozoa and oocyte both contribute to early embryonic resistance to heat shock is demonstrated. In addition, the use of reproductive biotechnologies to improve bovine thermotolerance, are outlined by comparing data from taurus, indicus and crossbred genotypes. PMID- 16246408 TI - Influence of repeated treadmill exercise on quality and freezability of stallion semen. AB - The objective of this study was to investigate changes of quality and freezability of stallion semen in response to repeated acute treadmill exercise. Ejaculates from 11 stallions were collected, evaluated and frozen weekly during four periods of 4 weeks each defined as before (period 1), during (period 2) and after (periods 3 and 4) intense exercise. In fresh semen the gel-free volume, sperm concentration, motility, normal sperm and sperm with major defects (acrosome defects, nuclear vacuoles, abnormal heads, midpiece defects and proximal droplets) were evaluated. In frozen-thawed semen, motility as well as viability (SYBR-14/PI) were examined. In period 2, all stallions were exercised on an indoor high speed treadmill twice a week (total of eight sessions) using an incremental workload test. Heart rate was monitored telemetrically during exercise and blood samples were taken for determination of cortisol, testosterone and lactate. Results of our investigation demonstrate that heart rate and the plasma concentrations of cortisol, testosterone and lactate significantly (P < 0.05) increased during each exercise session. Furthermore, significantly more major sperm defects were present in periods 3 (69.5+/-2.1%) and 4 (66.8+/-2.1%) than in periods 1 (62.2+/-2.4%) and 2 (62.5+/-2.2%). Acrosome defects increased towards the end of exercise but improved 3 weeks later to values observed before exercise. In frozen-thawed semen, motility was significantly lower in period 2 (45.4+/-2.3%) compared to period 4 (51.6+/-1.7%) and viability was significantly lower in period 2 (49.2+/-2.0%) than in periods 1 (53.8+/-2.1%) and 4 (53.7+/ 1.6%). Our results clearly demonstrate that in the stallion repeated strenuous treadmill exercise can negatively influence semen quality and freezability. PMID- 16246409 TI - Treatment with medications affecting dopaminergic and serotonergic mechanisms: effects on fluency and anxiety in persons who stutter. AB - Medications with dopamine antagonist properties, such as haloperidol, and those with serotonin reuptake inhibitor properties, such as clomipramine, have been shown to improve fluency. To examine the degree to which each of these two pharmacological mechanisms might independently affect fluency, a selective serotonin reuptake inhibitor, paroxetine, and a selective dopamine (D-2) antagonist, pimozide, were evaluated. Both types of medications also affect mood and anxiety, factors that could influence fluency levels. Therefore, we also evaluated the medications' effects on generalized and speech-related anxiety and the relationships between changes in anxiety and changes in fluency in 11 subjects with a history of developmental stuttering. The randomized, double blind, placebo-controlled crossover study that was designed had to be terminated prior to completion due to severe side effects following withdrawal from paroxetine. Even with a reduced sample size (n=6), significant improvement in percent fluent speaking time (p=0.02) was found using a telephone task between baseline and pimozide (n=6), with average duration of dysfluencies significantly shorter (p=0.04) but no significant difference in the estimated number of dysfluencies per minute. This significant improvement was associated with non significant increases in generalized anxiety, but non-significant decreases in speech-related anxiety. No significant differences were found in fluency between baseline and paroxetine (n=5). These preliminary results suggest that fluency improvement is more likely to be mediated by dopaminergic rather than serotonergic mechanisms. Due to its side effects, however, pimozide may be considered a risk for treatment of stuttering. EDUCATIONAL OBJECTIVES: As a result of reading this paper the reader will describe and explain: (1) how medications may affect fluency and the rationale for selecting medications for treatment trials; (2) the interrelationship between fluency and anxiety; and (3) factors important in developing clinical trials using medications. PMID- 16246410 TI - A comparative investigation of the speech-associated attitude of preschool and kindergarten children who do and do not stutter. AB - The data of recent research studies have shown that by 3 years of age children show an awareness of dysfluency and that by at least the age of six, youngsters who stutter have a speech-associated attitude that is more negative than that of their peers. These findings led to the present study in which the KiddyCAT, a self-report measure, was used to compare the attitude toward speech of 45 children, between the age of three and six, who stuttered with that of 63 who did not. The data of this investigation showed that, as a group, the preschool and kindergarten children who stuttered had significantly more in the way of a negative attitude toward their speech than was found among their non-stuttering peers of the same age and gender. This finding is not consistent with the classically held position that the reactive aspects of stuttering do not generally develop until well after its onset. It suggests the need to measure, by standardized means, the speech-associated attitude of incipient stutterers and, when appropriate, to make the assessment and treatment of negative attitude toward speech a meaningful aspect of therapy. EDUCATIONAL OBJECTIVES: The reader will be able to: (1) describe the difference in the speech-associated attitude of preschoolers and kindergartners who do and do not stutter; (2) summarize what we currently know about self-report tests used to assess speech-related attitude among children as young as 3; and (3) evaluate the usefulness of assessing the belief system of children whose fluency is considered problematic. PMID- 16246411 TI - A descriptive epidemiology of screen-based media use in youth: a review and critique. AB - The purpose of this systematic review was to (i) estimate the prevalence and dose of television (TV) viewing, video game playing and computer use, and (ii) assess age-related and (iii) secular trends in TV viewing among youth (< or = 18 yr). Ninety studies published in English language journals between 1949 and 2004 were included, presenting data from 539 independent samples (the unit of analysis). Results suggest contemporary youth watch on average 1.8-2.8 h of TV per day, depending on age and gender. Most (66%) are "low users" (< 2 h day(-1)) of TV but 28% watch more than 4 h day(-1). Boys and girls with access to video games spend approximately 60 and 23 min day(-1), respectively, using this technology. Computer use accounts for an additional 30 min day(-1). Age-specific data suggest TV viewing decreases during adolescence, but those considered "high users" at young ages are likely to remain high users when older. For children with access to a television set, the number of hours spent viewing does not appear to have increased over the past 50 years. PMID- 16246412 TI - Sabella spallanzanii filter-feeding on bacterial community: ecological implications and applications. AB - The filtration process of Sabella spallanzanii Gmelin on bacterial community was studied in a coastal area of the Northern Ionian Sea (Mediterranean Sea) at three sites, S. Vito, Lido Gandoli and Lido Silvana, where some specimens of S. spallanzanii were transplanted. Analyses were performed both on water and worm samples. A total of six microbial groups were examined: culturable heterotrophic bacteria, total culturable bacteria at 37 degrees C, culturable vibrios, total and fecal coliforms and fecal streptococci. The bacterial densities were usually orders of magnitude higher in the worm homogenates than in the corresponding seawater and the highest values were observed in August. The ability of S. spallanzanii to accumulate the microbial pollution indicators suggests this species can be employed as a bioindicator for monitoring water quality. Moreover, the accumulation capability of S. spallanzanii for specific micro-organisms provides a potential role in sewage bioremediation. PMID- 16246413 TI - Dual growth factor-induced angiogenesis in vivo using hyaluronan hydrogel implants. AB - Crosslinked hyaluronan (HA) hydrogels preloaded with two cytokine growth factors, vascular endothelial growth factor (VEGF) and keratinocyte growth factor (KGF), were employed to elicit new microvessel growth in vivo. As a major glycosaminoglycan (GAG) component of extracellular matrix (ECM), HA is an excellent biopolymeric building block for new biomimetic, biocompatible therapeutic materials. HA hydrogel film samples were surgically implanted in the ear pinnae of mice, and the ears were harvested at 7 or 14 days post implantation. Histologic analysis showed that each of the groups receiving an implant demonstrated significantly more microvessel density than control ears undergoing surgery but receiving no implant (p<0.001). Treatment groups receiving either co-delivery of both KGF and VEGF, an HA hydrogel lacking a growth factor or HA hydrogels containing a single cytokine were statistically unchanged with time, whereas treatment with KGF alone produced continuing increases in vascularization from day 7 to day 14. Strikingly, presentation of both VEGF and KGF in crosslinked HA generated intact microvessel beds with well-defined borders. In addition, an additive response to co-delivery of both cytokines in the HA hydrogel was observed. The HA hydrogels containing KGF+VEGF produced the greatest angiogenic response of any treatment group tested (NI=5.4 at day 14, where NI is a neovascularization index). This was 33% greater vessel density than in the next largest treatment group, that received HA+KGF (NI=4.0, p<0.002). New therapeutic approaches for numerous pathologies could be notably enhanced by the localized, sustained angiogenic response produced by release of both VEGF and KGF from crosslinked HA films. PMID- 16246414 TI - Magnesium and its alloys as orthopedic biomaterials: a review. AB - As a lightweight metal with mechanical properties similar to natural bone, a natural ionic presence with significant functional roles in biological systems, and in vivo degradation via corrosion in the electrolytic environment of the body, magnesium-based implants have the potential to serve as biocompatible, osteoconductive, degradable implants for load-bearing applications. This review explores the properties, biological performance, challenges and future directions of magnesium-based biomaterials. PMID- 16246415 TI - Inflammatory mediator release in normal bronchial smooth muscle cells is altered by pregnant maternal and fetal plasma independent of asthma. AB - Studies have shown that pregnancy can alter the pathophysiology of a pre-existing maternal disease such as asthma. However, the mechanisms that alter maternal asthma during pregnancy are presently unknown. Previous work has demonstrated that human bronchial smooth muscle (BSM) cells produce inflammatory factors in response to nonpregnant, atopic plasma. The aim of this study was to determine whether circulating pregnancy-derived factors in maternal and fetal plasma can stimulate inflammatory mediator release in BSM cells in the presence and absence of maternal asthma. Cultured human BSM cells were exposed to maternal and fetal plasma from normal pregnancies and pregnancies complicated by asthma. Inflammatory mediator release was determined by enzyme-linked immunosorbent assay (ELISA). Both maternal and fetal plasma from asthmatic and nonasthmatic individuals significantly increased production of interleukin (IL)-6 (ANOVA, P<0.001), regulated upon activation, normal T-cell expressed and secreted (RANTES) (ANOVA, P<0.01), and soluble intercellular cell-adhesion molecule-1 (sICAM-1) (ANOVA, P<0.01). There was no difference in inflammatory mediator release in response to asthma and nonasthmatic plasma. Eotaxin release was increased by pregnant asthmatic plasma (ANOVA, P<0.05). The results of this study suggest that circulating pregnancy-related factors can activate asthma-associated mediators in BSM cells. This change in BSM function may be one mechanism that contributes to increased asthma severity during pregnancy. PMID- 16246416 TI - Novel placental expression of 2,3-bisphosphoglycerate mutase. AB - 2,3-Bisphosphoglycerate mutase (2,3-BPGM), an erythroid-expressed enzyme, synthesises 2,3-bisphosphoglycerate (2,3-BPG), the allosteric modulator of haemoglobin. This ligand has a higher affinity for adult haemoglobin than for fetal haemoglobin and differential binding of it facilitates transfer of oxygen between adult and fetal blood by lowering the affinity of adult haemoglobin for oxygen. This paper reports the discovery that 2,3-BPGM is synthesised in non erythroid cells of the human placenta. Western blot analysis of placental extracts revealed high levels of 2,3-BPGM in the human placenta. Immunohistochemical staining and in situ hybridisation experiments indicated that abundant 2,3-BPGM is present in the syncytiotrophoblast layer of the placental villi at the feto-maternal interface. A cytochemical staining technique showed that the placental 2,3-BPGM is active, indicating that 2,3-BPG is synthesised in the outermost cells of the placenta. These observations demonstrate an unexpected and abundant presence of an enzyme key to oxygen release from adult haemoglobin, at the interface between maternal and fetal circulations. PMID- 16246417 TI - Arginine vasopressin does not contribute to seizures induced by intracerebroventricularly-injected pilocarpine. AB - Arginine vasopressin (AVP) has been shown to contribute to the production of seizures. Here, we aimed to investigate the effects of AVP on seizures induced by intracerebroventricular (i.c.v.) injection of pilocarpine. Rats were treated with 0.2-2.4 mg/5 microl pilocarpine intracerebroventricularly, to obtain the dose response relationship for behavioural seizures. 2.4 mg/5 microl pilocarpine induced status epilepticus in all rats and 0. 2 mg/5 microl pilocarpine did not produce any sign of seizure in any of the rats. In the second step, AVP (0.01 1000 ng/2 microl; i.c.v.) was injected 5 min before i.c.v. injection of a low dose pilocarpine (0.4 mg/5 microl) and rats were observed for percentage of status epilepticus, status epilepticus latency and behavioural seizure scores. None of the applied doses of AVP had any significant effect on seizures induced by 0.4 mg/5 microl i.c.v. pilocarpine. Subcutaneous injection of 1000 ng AVP 1h before 0.4 mg i.c.v. pilocarpine also did not produce significant difference with respect to the 0.4 mg pilocarpine group. Finally, pretreatment with neither an AVP V(1) receptor antagonist (25, 125, 250 ng/5 microl; i.c.v.) nor an AVP V(2) receptor antagonist (25, 125, 250 ng/5 microl; i.c.v.) prevented status epilepticus, induced by 2.4 mg/5 microl i.c.v. pilocarpine. We conclude that AVP does not act as a convulsant agent in centrally-induced pilocarpine seizures. PMID- 16246419 TI - Is there an association between ocular adnexal lymphoma and infection with Chlamydia psittaci? The University of Rochester experience. AB - Various subsets of extranodal marginal zone lymphomas of mucosa-associated lymphoid tissues (MALT lymphomas) have been associated with infectious organisms. Most notable of these is the association of gastric MALT lymphomas with Helicobacter pylori infection. In a recent publication Ferreri et al. [Ferreri AJ, Guidoboni M, Ponzoni M, De Conciliis C, Dell'Oro S, Fleischhauer K, et al. Evidence for an association between Chlamydia psittaci and ocular adnexal lymphomas. J Natl Cancer Inst 2004;96:586-94] reported the presence of C. psittaci DNA in 80% of 40 ocular adnexal lymphomas. Similar to the gastric MALT lymphoma data, a subset of these patients responded well to antibiotic treatment. We analyzed a set of ocular adnexal lymphomas and benign (non-neoplastic) lesions for evidence of C. psittaci DNA in patients from New York State. No evidence of C. psittaci DNA was seen in seven MALT-type ocular adnexal lymphomas, four non MALT ocular lymphomas, one Langerhans histiocytosis, and five reactive lymphoproliferations. We eliminated several possible reasons that would cause our study to fail to find C. psittaci DNA, including the presence of PCR inhibitors, inadequate template DNA, and sequence diversity in the target region in C. psittaci. The positive data were based primarily on patients from Italy, while our study involved only patients living in the Northeastern United States. This would suggest possible geographic differences in the etiology of ocular adnexal lymphomas. PMID- 16246418 TI - Granulocytic maturation in cultures of acute myeloid leukemia is not always accompanied by increased apoptosis. AB - In this issue of the Journal, Soucek et al. challenge the assumption that increased functional granulocytic maturation of HL-60, an ATRA-responsive acute myeloid leukemia cell line devoid of the APL-specific PML-RARalpha fusion protein, results in more rapid or more sustained cell death. In this model cell line, the authors demonstrate that TGFbeta1, a well-known haemopoietic growth factor, enhances retinoid-dependent cyto-differentiation and growth arrest while inhibiting apoptosis. Concomitantly, treatment of HL-60 cells with the combination of TGFbeta1 and the retinoid partially suppresses ATRA-dependent induction of TRAIL. This is a death receptor ligand of the TNF family implicated in the paracrine mechanism underlying the apoptotic action of ATRA in APL blasts The protein activates the death-receptor-dependent or extrinsic apoptotic pathway, which is associated with caspase-8 activation. Down-regulation of TRAIL is correlated to an increase in the levels of the anti-apoptotic c-FLIP(L) and Mcl-1 proteins that are likely to be involved in the suppression of caspase-8 activation and apoptosis. PMID- 16246420 TI - Histological changes in immune and endocrine organs of quail embryos: exposure to estrogen and nonylphenol. AB - Effects of 17beta-estradiol and p-nonylphenol were examined in the immune and endocrine organs of Japanese quail embryos. The test compounds were injected into the yolk of embryonated eggs. Injection of estrogen resulted in (1) disappearance of lymphoid cells and flattened development of plicae in the bursa of Fabricius, (2) decreased area of thyroid follicles and height of simple cuboidal epithelial cells in the thyroid, (3) increased follicular appearance of the thymus, and (4) development of an ovotestis in male embryos. Injection of nonylphenol did not induce flattened plicae in the bursa of Fabricius or development of an ovotestis in male embryos, but it increased the disappearance of lymphoid cells from the lymphoid follicles in the bursa, decreased the height of simple cuboidal epithelial cells surrounding the thyroid follicle, and increased the follicle like structure in the thymus in male embryos. These results suggest that nonylphenol has estrogenic effects, but these are low compared to those of estrogen itself. PMID- 16246421 TI - Parkinson's disease, chronic hydrocarbon exposure and striatal neuronal damage: a 1-H MRS study. AB - Several patients with Parkinson' s disease (PD) reveal an history of chronic exposure to hydrocarbon-solvents. Chronic exposure to hydrocarbon-solvents has been proposed as a risk factor for more severe forms of PD with earlier onset of symptoms and reduced response to dopaminergic therapy. A direct correlation between disease severity and exposure degree has been previously shown. Seven exposed PD patients (two with low degree exposure and five with high degree exposure), 10 unexposed PD patients matched for sex, age and Hoehn and Yahr scale (=3 in the "on" phase), and 10 unexposed PD patients matched for sex, age and l dopa daily intake instead of disease severity (Hoehn and Yahr scale=3.5 in the "on" phase) were studied. Twenty normal subjects without previous exposure to hydrocarbon-solvents and matched for age and sex with HPD patients were studied for comparison. The purpose of the study was to assess neuronal degeneration in the striatum of exposed vs unexposed PD patients. The authors investigated whether neuronal damage/loss was detectable in the lentiform nucleus measuring N acetylaspartate (NAA) levels by 1-H MRS. Multiple single voxel MRS water suppressed spectra were obtained also from the white matter and the occipital lobe. NAA was normal in the lentiform nucleus of patients with low exposure as well as in patients with no exposure whereas it was decreased in PD patients with high degree exposure. White matter and occipital lobe NAA content was normal both in exposed and unexposed PD patients. Clinical expression is more severe in PD patients with previous high degree solvent exposure because of the associated post-synaptic damage of the nigro-striatal pathway. PMID- 16246422 TI - Epstein-Barr virus infected natural killer cell lymphoma in a patient with hypersensitivity to mosquito bite. AB - Hypersensitivity to mosquito bite (HMB) can occur in association with chronic Epstein-Barr virus (EBV) infection and natural killer (NK) cell leukaemia/lymphoma, which was named 'Tokura-Ishihara disease'. This disease is very rare and most previous reports have been documented in Japan. We present a patient who suffered from of pustules on skin, high fever, myalgia and multiple lymph node enlargements after mosquito bite from childhood. Recently, multiple lymph nodes were palpable on his both inguinal area. Peripheral blood smear (PBS) revealed many large granular lymphocytes and the skin lesion showed a dense dermal and subcutaneous infiltrate of lymphocytes. The lymph nodes and perinodal adipose tissue were infiltrated by atypical lymphoid cells in which EBER-positive signals were identified by in situ hybridization using EBV encoded RNA-1 probe. He was diagnosed as having Tokura-Ishihara disease and receives chemotherapy now. Here, we report a case of this disease with a precise pathological description on the lymph node biopsy. PMID- 16246423 TI - Legionella pneumophila pneumonia during pregnancy: a case report. AB - Legionella pneumophila pneumonia during pregnancy can have serious consequences for the mother and lead to fetal distress. We report a case of L. pneumophila pneumonia in a pregnant woman at 31 weeks gestation. With early diagnosis and appropriate treatment, the outcome was favorable with delivery of a healthy infant at 40 weeks gestation. PMID- 16246424 TI - Respiratory cycle as time basis: an improved method for averaging olfactory neural events. AB - In the mammalian olfactory system, neural activity appears largely modulated by respiration. Accurate analysis of respiratory synchronized activity is precluded by the variability of the respiratory frequency from trial to trial. Thus, the use of respiratory cycle as the time basis for measuring cell responses was developed about 20 years ago. Nevertheless, averaging oscillatory component of the activity remains a challenge due to their rhythmic features. In this paper, we present a new respiratory monitoring setup based on the measurement of micropressure changes induced by nasal airflow in front of the nostril. Improvements provided by this new monitoring setup allows automatic processing of respiratory signals in order to extract each respiratory period (expiration and inspiration). The time component of these periods, which can differ from trial to trial, is converted into a phase component defined as [-pi, 0] and [0, pi] for inspiration and expiration, respectively. As opposed to time representation, the phase representation is common to all trials. Thus, this phase representation of the respiratory cycle is used as a normalized time basis permitting to collect results in a standardized data format across different animals and providing new tools to average oscillatory components of the activity. PMID- 16246425 TI - Improved detection of fluorogold-labeled neurons in long-term studies. AB - Fluorogold (FG) is a widely used neuroanatomical tracer. However, because FG labeled neurons become undetectable over time, its use has been limited in long term studies. We investigated whether the detection of FG in retrogradely labeled neurons in long-term studies can be improved by immunohistochemistry (IHC) using an antibody to FG. We performed intraperitoneal injections of a FG solution to retrogradely label all parasympathetic preganglionic neurons (PPNs) and motoneurons (MNs) in the S1 spinal cord segment in adult rats. At 1, 6, and 12 weeks after the tracer injection, sections were immunohistochemically processed for FG and choline acetyltransferase (ChAT), an endogenous marker for all PPNs and MNs. Stereological counts demonstrated no cell loss of FG-labeled PPNs and MNs at 6 and 12 weeks. Cell size measurements showed that FG-immunolabeled neurons were smaller at 12 weeks, but not at 6 weeks. However, it is likely that there was no neuronal atrophy, but loss/degradation of the dye at a timepoint between 6 and 12 weeks, as ChAT-immunolabeled neurons showed no cell size reduction at 12 weeks. Our results suggest that the use of an antibody against FG improves the detection of FG for reliable neuronal counts and that the dye is not toxic to the retrogradely labeled neurons. We conclude that FG-labeling is a useful tool to determine neuronal counts in long-term studies, but should be used cautiously for neuronal size measurements. PMID- 16246426 TI - Fluorescent labeling of rat auditory brainstem circuits for synaptic and electrophysiological studies. AB - We visualized neurons with fluorescent agents, both retrogradely and anterogradely, to identify the input and output neuronal pathways in the rat auditory system. Output neurons of dorsal cochlear nucleus (DCN) were labeled retrogradely by injecting fluorescent microspheres into the inferior colliculus. Electrical recordings were made from the labeled fusiform cells of DCN with the whole-cell patch-clamp recording technique in slice preparations. DiI and Sindbis virus expressing membrane-targeted green fluorescent protein (GFP) were adopted for anterograde labeling. Auditory nerve fibers (ANFs) were labeled by injecting DiI into the cochlea, and the contralateral projection to the medial nucleus of the trapezoid body (MNTB) by injecting DiI and GFP into the ventral cochlear nucleus. A single ANF projecting to a DCN fusiform cell was electrically stimulated by a glass electrode and EPSCs were recorded using whole-cell patch clamp recording methods. EPSCs were sensitive to the positioning of the electrode, and the size of EPSCs was constant irrespective of stimulus intensity, indicating that a single fiber was stimulated. Large EPSCs were generated from MNTB principal cells by stimulating the labeled fiber with a Calyx of Held terminal. The membrane excitability and EPSCs recorded after fluorescence labeling were similar to those previously reported. We confirm that the fluorescence labeling is effective to visualize neural networks and is useful to investigate the electrophysiological properties of neurons and synapses. PMID- 16246427 TI - Cryopreservation of brain mitochondria: a novel methodology for functional studies. AB - Often, comparative studies involving large number of animals or human post-mortem tissue samples are precluded, especially those requiring structurally and functionally intact cells and/or organelles. The ability to 'bank' such samples for storage and restore or 'reanimate' them at a later time without causing damage to the structure and/or function becomes imperative. However, to date, such attempts have produced conflicting results. We here demonstrate for the first time that isolated rat brain mitochondria can be successfully cryopreserved and restored for later use. We added a well characterized cryoprotectant 10% (v/v) dimethyl sulfoxide (DMSO) to purified rat cortical mitochondria and allowed them to cool at a uniform rate of approximately 1 degree C/min and stored them at -80 degrees C. Freshly isolated as well as reanimated brain mitochondria were analyzed for respiration. Structural integrity of cryopreserved mitochondria was also verified by electron microscopy. Mitochondrial membrane marker levels were assessed along with cytochrome c levels. Intact structure and function of the cryopreserved brain mitochondria observed allows us the opportunity to store mitochondria for longer periods of time as well as perform metabolic studies as needed. This will considerably expand the time-frame required for carrying out functional analysis in large comparative studies. PMID- 16246428 TI - A comparison of cell transplantation and retroviral gene transfection as tools to study lineage and differentiation in the rat spinal cord. AB - Establishing the cell lineage relationships of cells during development allows insight into when and where developmental decisions are made. In the developing spinal cord, the origin and fate of radial glial has yet to be determined. One way in which to address this question is to transplant enriched populations of radial glia into the ventricular zone (VZ) region of host embryos to examine the lineage and differentiation pattern of these cells. An indirect selection procedure using immunomagnetic beads (Dynabeads; Dynal Biotech) was used here to isolate spinal cord radial glia. This negative immunoselection procedure resulted in a high yield of radial glia. A fluorescent cytoplasmic dye (Cell Tracker Green CMFDA) was used to label radial glia before transplantation. The role of radial glia as progenitor cells can also be examined using a green fluorescent protein (GFP)-expressing retroviral vector. The retroviral vector allows dividing cells in the VZ region of the spinal cord to be tracked by labelling them with GFP. Both techniques were utilised here to successfully label and examine embryonic spinal cord radial glia in vivo after a microinjection of either fluorescently labelled radial glia or replication-incompetent GFP-expressing retrovirus in utero. PMID- 16246429 TI - Phenotyping of lymphocytes expressing regulatory and effector markers in infiltrating ductal carcinoma of the breast. AB - Dysfunction of the host immune system in cancer patients can be due to a number of reasons including suppression of tumour associated antigen reactive lymphocytes by regulatory T (Treg) cells. In this study, we used flow cytometry to determine the phenotype and relative abundance of the tumour infiltrating lymphocytes (TILs) from 47 enzymatically dissociated tumour specimens from patients with infiltrating ductal carcinoma (IDC) of the breast. The expression of both effector and regulatory markers on the TILs were determined by using a panel of monoclonal antibodies. Analysis revealed CD8(+) T cells (23.4+/-2.1%) were predominant in TILs, followed by CD4(+) T cells (12.6+/-1.7%) and CD56(+) natural killer cells (6.4+/-0.7%). The CD4(+)/CD8(+) ratio was 0.8+/-0.9%. Of the CD8(+) cells, there was a higher number (68.4+/-3.5%) that expressed the effector phenotype, namely, CD8(+)CD28(+) and about 46% of this subset expressed the activation marker, CD25. Thus, a lower number of infiltrating CD8(+) T cells (31.6+/-2.8%) expressed the marker for the suppressor phenotype, CD8(+)CD28(-). Of the CD4(+) T cells, 59.6+/-3.9% expressed the marker for the regulatory phenotype, CD4(+)CD25(+). About 43.6+/-3.8% CD4(+)CD25(+) subset co-expressed both the CD152 and FOXP3, the Treg-associated molecules. A positive correlation was found between the presence of CD4(+)CD25(+) subset and age (> or =50 years old) (r=0.51; p=0.045). However, no significant correlation between tumour stage and CD4(+)CD25(+) T cells was found. In addition, we also found that the CD4(+)CD25(-) subset correlated with the expression of the nuclear oestrogen receptor (ER)-alpha in the tumour cells (r=0.45; p=0.040). In conclusion, we detected the presence of cells expressing the markers for Tregs (CD4(+)CD25(+)) and suppressor (CD8(+)CD28(-)) in the tumour microenvironment. This is the first report of the relative abundance of Treg co-expressing CD152 and FOXP3 in breast carcinoma. PMID- 16246430 TI - A pilot project on hospital-based universal newborn hearing screening: lessons learned. AB - OBJECTIVE: This paper reports the performance of a newly implemented hospital based universal newborn hearing screening programme and the challenges to the effective implementation. METHODS: Data of 4437 babies screened from April 2003 to February 2004 at Hospital Universiti Kebangsaan Malaysia was analyzed to measure the performance of the newborn hearing screening programme. Quality indicators, which include the coverage rate, initial refer rate, return for follow-up rate, ages of diagnosis and intervention were calculated. Factors contributing to poor compliance for follow-up were examined through questionnaire survey of 341 parents who did not bring their babies for the initial screening follow-up. RESULTS: The findings of this study revealed unsatisfactory performance of the hearing screening programme compared to the Joint committee of Infant Hearing recommendation [Joint Committee on Infant Hearing Year 2000 Position Statement: Principles and Guidelines for Early Hearing Detection and Intervention Programs, Pediatrics 106 (4) (2000) 798-817]. The coverage rate, initial refer rate, and return for follow-up rate were 84.64, 11.97 and 56.97%, respectively. The average age of diagnosis was 3.56 months (S.D. 1.33). Only 1 of the 16 babies identified as having hearing loss through the screening programme has been fitted with hearing aids. Delay in coming to audiological certainty was one of the reasons hampering early intervention of these children. The commonly cited reasons for not returning for screening follow-up reflect the need to create public awareness of the importance of early diagnosis and intervention of hearing loss. CONCLUSIONS: The implementation of the present screening programme needs to be reviewed. Factors contributing to its unsatisfactory performance must be identified and steps must be taken to resolve them so that early identification and intervention of permanent congenital hearing loss can become a reality. PMID- 16246431 TI - Central giant cell reparative granuloma of the mandible caused by a molar tooth extraction: special reference to the maneuver of drilling the surgical field. AB - Central giant cell reparative granuloma (CGCRG) is an uncommon benign, reactive osseous lesion usually located in the mandible and maxilla. Although it is histologically benign, it may be locally destructive. There is still controversy on its development and growth pattern. Surgery is the treatment traditionally recommended. In this article, we presented a 12 year-old girl CGCRG of the mandible caused by a molar tooth extraction and discussed its histopathological, clinical, radiological and therapeutic features in the light of the current literature. Also we described our additional surgical maneuver 'drilling the surgical field' after the removal of the lesion. PMID- 16246432 TI - Universal newborn hearing screening follow-up in two Georgia populations: newborn, mother and system correlates. AB - BACKGROUND: Nearly half of babies who "fail" their newborn hearing screening do not receive appropriate follow-up. Various explanations have been suggested. OBJECTIVE: To investigate, in two contrasting populations of newborns in Georgia, contemporaneous medical, socio-demographic and screening correlates of follow-up after newborn hearing screening. Three hypotheses were addressed: (1) follow-up correlated with particular medical, social and demographic features; (2) screening performance indicators correlated with follow-up; and (3) screening policies and procedures correlated with follow-up. METHODS: The studied babies, born July 2001 through June 2003 at Atlanta's Piedmont Hospital and Waycross Health District, had failed screening in both ears. Each site had about 4000 births per year, and a plan for outpatient follow-up of in-patient "fail" babies. RESULTS: At Piedmont, where Medicaid funded less than 7% of the births, 57% (130/230) had known follow-up to outpatient screening. In contrast, at Waycross, where Medicaid funded 89.6% of the births, 100% (38/38) had follow-up. The first study hypothesis was only partially supported by the data-from Piedmont, but not Waycross. White race and maternal age 30 years or older correlated with following up: odds ratios 2.07 (95% CI 1.17, 3.68) and 1.83 (1.05, 3.17), respectively. Private health insurance and marriage trended with following-up. Follow-up rates did not correlate with the rates of screening refusal, missed screening and "fail" rates. Follow-up was unrelated to mothers witnessing the screening, receiving in-person explanations and having appointments arranged for them. The Piedmont and Waycross programs were quite different. Piedmont's began in 2001, mothers typically learned about screening when it was done and "fail" babies were reported to many public health entities. In contrast, the Waycross program was more than 20 years old, prenatal care included teaching about screening and reporting was to one public health entity. CONCLUSIONS: Though maternal socio demographic features (poor, non-white, young) and access problems have been considered factors for non-follow-up, this study found two program characteristics most important: the lack of prenatal education about newborn hearing screening, and the lack of functionally integrated hospital hearing information with Public Health. PMID- 16246433 TI - Adaptation of nasometry to Hungarian language and experiences with its clinical application. AB - OBJECTIVES: (1) To adopt the nasometry for the Hungarian language and to obtain normative nasalance scores. (2) To compare our results with the data of other languages and to evaluate the correlation between nasalance scores and perceptual ratings of nasality. (3) To use the nasometry in various fields of the otolaryngological, phoniatric, and logopedic diagnostics, therapy and documentation. METHODS: (1) To determine the normative nasalance scores regarding the Hungarian language, we included 30 children aged 5-7 years and 45 adults in the 20-25 years age group. In the latter group 15 subjects were speech therapists and 30 phonetically untrained people-15 males and 15 females. STUDY DESIGN: phonation of isolated vowels, articulation of spirants, cyclical repetition of affricates, pronunciation of various (oral, nasal, mixed type) sentences and evaluation of the nasalance score in continuous speech. (2) Thirty-six persons (12 speech pathologists, 12 logopedic students, 12 phonetically uneducated individuals) evaluated the children's physiological and nasal speech recordings with a 3-point scale. (3) Two hundred and forty-eight children of kindergarten age were examined, 20 infants and 6 adult singers in the following fields: evaluation of hypernasality due to cleft palate or velopharyngeal insufficiency (VPI), and of the success of the therapy; examination of hyponasality in cases of enlarged adenoid and allergic rhinitis; evaluation of the speech of hard-of hearing people; differentiation between nasal sigmatism and hyperrhinophony; testing of the resonance in professional singers; examination of infant cry; application of nasometry in the therapy. RESULTS: The mean value of the nasalance score using the oral sentence: "Zsuzsi kutyaja ugat" is 11-13%, in the nasal sentence ("A majom banant enne") 56%, while that of the mixed sentence representing the Hungarian language ("Jo napot kivanok!") falls in the 30-40% range. The resonance grows with aging and there is no significant difference between genders. The nasalance score is greater with phonetically trained people. Our data correlate with the values of other languages. The correlation is significant between the nasalance scores and perceived nasality (r=0.901). Practical results: Values above 40% in cases of VPI using mixed sentences may support the indication of velopharyngoplasty, together with the subjective evaluation of nasality and other tests. In cases with rhinitis and adenoid vegetation the nasalance score remains below 20%. The nasality value is increased in sensorineural hearing loss, and is decreased in cases with conducting hearing impairment. In nasal sigmatism not the vowels' but the nasality of consonants grow. The difference between the nasalance score of the cry in clefted and non cleft infants is significant (26% versus 36%): this observation could give possibility in the future to screen babies with congenital hearing problems or hidden VPI. Alterations in nasalance can be documented with nasometry in professional singers when they increase the nasal resonance to grow the power capacity of their voice. The nasometry procedure is a significant help also in speech therapy through the real time visual and auditive control. CONCLUSIONS: The otolaryngological, phoniatric and logopedic diagnostics and therapy is significantly widened with nasometry which is a quick, non-invasive and objective procedure, measuring the nasal resonance of the speech. PMID- 16246434 TI - Antidepressant action: to the nucleus and beyond. AB - After decades of effort, the field of depression research is far from understanding how antidepressant drugs mediate their clinical effects. The time lag of 2-6 weeks of therapy that is necessary to obtain antidepressant efficacy indicates a requirement for long-term regulation of molecules activated by drug treatment. The focus of antidepressant research has thus expanded from examining acute monoamine-mediated mechanisms to include long-term transcriptional regulators such as cAMP response element-binding protein (CREB) and trophic factors such as brain-derived nerve growth factor and insulin-like growth factor. In addition, the recent discovery of antidepressant-induced neurogenesis provides another avenue by which antidepressants might exert their effects. Current efforts are aimed at understanding how CREB and trophic factor signaling pathways are coupled to neurogenic effects and how alterations in behavioral, molecular and cellular endpoints are related to the alleviation of the symptoms of depression. PMID- 16246435 TI - Glia: novel counter-regulators of opioid analgesia. AB - Development of analgesic tolerance and withdrawal-induced pain enhancement present serious difficulties for the use of opioids for pain control. Although neuronal mechanisms to account for these phenomena have been sought for many decades, their bases remain unresolved. Within the past four years, a novel non neuronal candidate has been uncovered that opposes acute opioid analgesia and contributes to development of opioid tolerance and tolerance-associated pain enhancement. This novel candidate is spinal cord glia. Glia are important contributors to the creation of enhanced pain states via the release of neuroexcitatory substances. New data suggest that glia also release neuroexcitatory substances in response to morphine, thereby opposing its effects. Controlling glial activation could therefore increase the clinical utility of analgesic drugs. PMID- 16246436 TI - Differential behavioral and neurochemical effects of cocaine after early exposure to methylphenidate in an animal model of attention deficit hyperactivity disorder. AB - We used a putative animal model of attention deficit hyperactivity disorder (ADHD), the SHR rat, to examine the effects of repeated exposure to methylphenidate (MPH; Ritalin) during the pubertal period on cocaine-induced conditioned place preference and dopamine (DA) levels in the nucleus accumbens (NAc) in adulthood. Our results indicate that early exposure to methylphenidate diminishes sensitivity to the incentive properties of cocaine in adulthood, but it does so without altering the response of the mesolimbic dopamine system. PMID- 16246437 TI - Reduced locomotion in the serum and glucocorticoid inducible kinase 3 knock out mouse. AB - The serum and glucocorticoid inducible kinase isoform SGK3 is expressed in the brain including hippocampal neurons. It is activated by phosphoinositide-3 (PI3) kinase and thus a putative target of neurotrophic factors. In vitro experiments pointed to the ability of SGK3 to regulate several transporters and ion channels including the AMPA receptor GluR1. In order to explore the in vivo functional significance of SGK3 in the regulation of spatial learning and exploratory behavior, we assessed the performance of SGK3 knockout mice (SGK3-/-) and their wild type littermates (SGK3+/+) in a place navigation task in the water-maze, radial maze in a battery of forced and free exploration tests, acoustic startle and a test for motoric coordination. According to water-maze and radial maze testing reference and working memory was intact in SGK3-/- mice. However, detailed analysis of swimming patterns of SGK3-/- mice in the water-maze revealed a deficit in precision and goal-directed navigation in space. SGK3-/- mice showed reduced exploratory activity, which was observed in several environments and increased centre field avoidance in the open-field. SGK3-/- mice further showed reduced darting behavior on open surfaces, indicating that the knock out may modify basic patterns of locomotion. In conclusion, lack of SGK3 leads to subtle behavioral defects which may result from deranged neuronal regulation of transporters and ion channels. PMID- 16246438 TI - Cloning and characterization of a high mobility group box 1 (HMGB1) homologue protein from Schistosoma mansoni. AB - Mammalian homologue of high mobility group box chromatin protein (HMGB) 1 was identified and cloned from human parasites, Schistosoma mansoni and S. haematobium. Sequence analyses showed that the parasite HMGB1s has 35-40% identity to human and rodent HMGB1s, and 33% identity to Caenorhabditis elegans HMGB1. Parasite HMGB1s also contains an A box and B box domain similar to mammalian HMGB1, however, it lacks the C-terminal tail that is present in mammalian HMGB1s. Analysis of the expression of HMGB1 in various life cycle stages of S. mansoni reveal S. mansoni HMGB1 (SmHMGB1) as a stage-specific protein, expressed abundantly in egg and adult female stages and at moderate levels in skin-stage schistosomula. Significant levels of SmHMGB1 were also present in excretory secretions of egg stages. Subsequent characterization studies showed that SmHMGB1 is a potent inducer of pro-inflammatory cytokines such as TNFalpha, IL-1Ralpha, IL-2Ralpha, IL-6, IL-13, IL-13Ralpha1, IL-15 and MIP-1alpha from mouse peritoneal macrophages. Pro-inflammatory activity, especially production of TNFalpha-inducing activity, appears to be a function of the B box domain protein. This was confirmed by both real-time reverse transcription PCR and by cytokine ELISA. Thus, results presented in this study suggest that SmHMGB1 may be a key molecule in the development of host inflammatory immune responses associated with schistosomiasis. PMID- 16246440 TI - Intravenous mesenchymal stem cell therapy early after reperfused acute myocardial infarction improves left ventricular function and alters electrophysiologic properties. AB - Direct intramyocardial injection of mesenchymal stem cells (MSCs) improves left ventricular ejection fraction (LVEF) and may increase ventricular arrhythmia in hearts with myocardial infarction (MI). We hypothesized that intravenous MSCs given early after acute MI would engraft in injured myocardium, improve LV function, and result in pro-arrhythmic electrical remodeling. We created an apical infarction in swine by balloon occlusion/reperfusion, administered diI labeled allogeneic bone marrow derived MSCs intravenously 30 min post-reperfusion and measured LVEF and wall thickness at baseline, 1 month, and 3 months. Epicardial effective refractory periods (ERPs) were determined before sacrifice. At 3 months, treated pigs [n=7] had significantly higher LVEF than controls [n=8] (49+/-2% vs. 44+/-3%, P=0.015) and significantly less wall thickening of non infarcted myocardium. ERPs were significantly shorter than controls at all pacing cycle lengths (P20 microg/min). No significant change in GFR from baseline after 16 weeks was observed for either treatment group. These data provide reassurance that type 2 diabetic patients can be treated with higher efficacy statins without clinically meaningful effects on urinary albumin excretion. PMID- 16246449 TI - Resistance to adefovir: a new challenge in the treatment of chronic hepatitis B. PMID- 16246450 TI - NAFLD treatment: cognitive-behavioral therapy has entered the arena. PMID- 16246451 TI - Pros and cons of TIPS for refractory ascites. PMID- 16246452 TI - Keeping cool in acute liver failure: rationale for the use of mild hypothermia. AB - Encephalopathy, brain edema and intracranial hypertension are neurological complications responsible for substantial morbidity/mortality in patients with acute liver failure (ALF), where, aside from liver transplantation, there is currently a paucity of effective therapies. Mirroring its cerebro-protective effects in other clinical conditions, the induction of mild hypothermia may provide a potential therapeutic approach to the management of ALF. A solid mechanistic rationale for the use of mild hypothermia is provided by clinical and experimental studies showing its beneficial effects in relation to many of the key factors that determine the development of brain edema and intracranial hypertension in ALF, namely the delivery of ammonia to the brain, the disturbances of brain organic osmolytes and brain extracellular amino acids, cerebro-vascular haemodynamics, brain glucose metabolism, inflammation, subclinical seizure activity and alterations of gene expression. Initial uncontrolled clinical studies of mild hypothermia in patients with ALF suggest that it is an effective, feasible and safe approach. Randomized controlled clinical trials are now needed to adequately assess its efficacy, safety, clinical impact on global outcomes and to provide the guidelines for its use in ALF. PMID- 16246453 TI - Regulation of two rat mas-related genes in a model of neuropathic pain. AB - The mas-related gene (Mrg) family is a large family of G-protein-coupled receptors which are variable in number depending on species. The so-called sensory-neuron-specific receptors (SNSRs) make up a subset of the Mrg family, and several of these have been implicated in nociceptive processes. To verify their specific localization in sensory ganglia, we have determined the expression patterns of two of them, rMrgA and rMrgC, in a panel of rat tissues. The quantitative PCR results in the rat tissue panel indicate that, while several non neuronal tissues contain significant levels of mRNA for both receptors, these two receptors are most highly expressed in dorsal root ganglia and trigeminal ganglia. Given this, we have examined the effects of spinal nerve ligation (SNL) on the expression of these genes. Peripheral neuropathy induced by ligation of spinal nerves at L5 and L6 resulted in a pronounced mechanical allodynia. These behavioral changes in tactile sensitivity were accompanied by significant decreases (10- to 100-fold) in the mRNA expression of both rMrgA and rMrgC exclusively in the L5 and L6 dorsal root ganglia ipsilateral to the SNL. In situ hybridization studies demonstrated that this decrease did not result from neuronal loss but rather from a reduction in the hybridization signals for rMrgC over small-to-medium diameter L5 and L6 dorsal root ganglia neurons. While the functional implications of the altered regulation of rMrgA and rMrgC in neuropathic pain models remain unclear, the results suggest that therapeutics targeting these receptors may have limited utility. PMID- 16246454 TI - Effect of water deprivation on aquaporin 4 (AQP4) mRNA expression in chickens (Gallus domesticus). AB - Aquaporin (AQP) 4 is a member of the AQP gene family of water-selective transport proteins. We studied the effect of water deprivation on AQP4 gene expression in chickens. The nucleotide sequence of a chicken aquaporin 4 (AQP4) cDNA that encodes a protein of 335 amino acids showed high homology to mammalian AQP4. Using Northern blotting analysis, AQP4 mRNA in chickens was observed as a band of approximately 5.5 kb in several tissues in addition to the hypothalamus, proventriculus, kidney, and breast muscle. Quantitative analysis by real-time RT PCR analysis showed that the mRNA expression of AQP4 in the hypothalamus significantly increased after dehydration. On the other hand, the mRNA expression of AQP4 in the kidney significantly decreased after dehydration. This suggests that AQP4 may play a pivotal role in osmoregulation in the chicken brain. PMID- 16246455 TI - A calcium binding protein, calbindin-D9k, is mainly regulated by estrogen in the pituitary gland of rats during estrous cycle. AB - As a member of family of cytosolic calcium binding proteins, Calbindin-D9k (CaBP 9k) is expressed in female reproductive system and regulated by steroid hormones, estrogen (E2) and progesterone (P4), but its expression and role in pituitary gland have not been elucidated yet. Thus, in this study, we elucidated the expression of CaBP-9k mRNA and protein in pituitary gland of rats. During estrous cycle of rats, pituitary CaBP-9k level fluctuated, and its mRNA was highly elevated during an E2-dominant stage (proestrus and estrus), whereas its level disappeared at a P4-dominant stage (metestrus and diestrus). In parallel with CaBP-9k mRNA, an increased level of CaBP-9k protein was observed during proestrus and estrus, suggesting that pituitary CaBP-9k may be up-regulated by E2. In addition, spatial CaBP-9k expression was attested by immunohistochemistry. Pituitary CaBP-9k protein was localized in the cytoplasm of a specific cell type in the anterior lobe, and the positive cells were abundant at proestrus and estrus. The CaBP-9k-positive cells were mainly localized in the acidophils producing growth hormones and prolactin. To verify hormonal regulation of pituitary CaBP-9k in this tissue, immature rats were treated with a physiological dose of E2 in the absence or presence P4 for 3 days. In a time-dependent experiment, pituitary CaBP-9k protein was induced at 48 h after the final E2 injection. A significant increase in CaBP-9k protein was caused by E2, whereas P4 antagonized E2-stimulated CaBP-9k expression as similarly observed in the uterus. Taken together, these results indicated for the first time that pituitary CaBP-9k expression is regulated during estrous cycle, and its expression is mainly controlled by E2 and antagonized by P4, suggesting that pituitary CaBP-9k in female rats may be involved in the central function of the reproduction system. PMID- 16246456 TI - The high and middle molecular weight neurofilament subunits regulate the association of neurofilaments with kinesin: inhibition by phosphorylation of the high molecular weight subunit. AB - Kinesin participates in axonal transport of neurofilaments (NFs), but the mode by which they attach to kinesin is unclear. We compared the association of NFs with kinesin in mice expressing or lacking NF-H or NF-M. In normal and M-/- mice, the leading edge of metabolically labeled NF subunits was selectively co-precipitated with kinesin. By contrast, the entire wave of radiolabeled subunits co precipitated with kinesin in H-/- mice. Similar bulk levels of NFs co precipitated with kinesin from normal and H-/- mice, but reduced levels co precipitated from M-/- mice. These data suggest that both NF-H and NF-M regulate the association of NFs with kinesin. They further indicate that phosphorylation of NF-H dissociates NFs from kinesin and provides a mechanism by which NF-H phosphorylation can contribute to the slowing of NF axonal transport. PMID- 16246457 TI - Caytaxin deficiency causes generalized dystonia in rats. AB - The genetically dystonic rat (SD-dt:JFL) is an autosomal recessive model of generalized dystonia. Without cerebellectomy, the dt rat dies prior to Postnatal Day 40. The dt locus was mapped to a 4.2 Mb region on Chr 7q11 and candidate genes were screened with semi-quantitative RT-PCR. Then, Southern blotting and genomic DNA sequencing identified the 3'-long terminal repeat portion of an intracisternal A particle element inserted into Intron 1 of Atcay, the gene which encodes caytaxin. Northern and Western blotting and quantitative real-time RT-PCR defined the Atcay allele in dt rats (Atcay(dt)) as hypomorphic. To establish a framework for functional studies of caytaxin, the developmental expression of rat Atcay transcript was analyzed with Northern blotting, relative quantitative multiplex real-time RT-PCR (QRT-PCR) and in situ hybridization. With a multiple tissue Northern blot, three Atcay transcripts were identified in brain but none were present in heart, spleen, lung, liver, muscle, kidney or testis. With a multiple time-point Northern blot, the same three transcripts were present in cerebellum at Embryonic Day (E15), Postnatal Day 1 (P1), P7, P14, P36 and 8 months. During early development (E15 to P14), the relative proportion of the smallest transcript was increased. QRT-PCR was performed with total RNA from cerebral cortex, striatum, thalamus, hippocampus and cerebellum. Transcript levels peaked at P7 in hippocampus, increased linearly from P1 to P36 in cerebellum, and showed minimal developmental regulation in cerebral cortex. Radioactive in situ hybridization localized Atcay transcript to seemingly all neuronal populations in brain. In cerebellum, Atcay transcript was present in the molecular, Purkinje and granular layers; transcript density in the molecular layer peaked at P14. In the background of previous biochemical, behavioral and electrophysiological studies in the dt rat, our data are compatible with a vital role for caytaxin in the development and neurophysiology of cerebellar cortex. PMID- 16246458 TI - Analysis of anaerobic BTX biodegradation in a subarctic aquifer using isotopes and benzylsuccinates. AB - In situ biodegradation of benzene, toluene, and xylenes in a petroleum hydrocarbon contaminated aquifer near Fairbanks, Alaska was assessed using carbon and hydrogen compound specific isotope analysis (CSIA) of benzene and toluene and analysis of signature metabolites for toluene (benzylsuccinate) and xylenes (methylbenzylsuccinates). Carbon and hydrogen isotope ratios of benzene were between -25.9 per thousand and -26.8 per thousand for delta13C and -119 per thousand and -136 per thousand for delta2H, suggesting that biodegradation of benzene is unlikely at this site. However, biodegradation of both xylenes and toluene were documented in this subarctic aquifer. Biodegradation of xylenes was indicated by the presence of methylbenzylsuccinates with concentrations of 17-50 microg/L in three wells. Anaerobic toluene biodegradation was also indicated by benzylsuccinate concentrations of 10-49 microg/L in the three wells with the highest toluene concentrations (1500-5000 microg/L toluene). Since benzylsuccinate typically accounts for a very small fraction of the toluene present in groundwater (generally <1 mol%), the signature metabolite approach works best at higher toluene concentrations when it is not constrained by detection limits. In wells with lower toluene concentrations (410-640 microg/L), carbon and hydrogen isotopic values were enriched by up to approximately 2 per thousand for delta13C and approximately 70 per thousand for delta2H. This evidence of isotopic fractionation verifies the effects of biodegradation in these low concentration wells where metabolites may already be below detection limits. The combined use of signature metabolite and CSIA data is particularly valuable given the challenge of verifying biodegradation in subarctic environments where degradation rates are typically much slower than in temperate environments. PMID- 16246459 TI - Combined use of field and laboratory testing to predict preferred flow paths in an heterogeneous aquifer. AB - Elevated nitrate concentrations within a municipal water supply aquifer led to pilot testing of a field-scale, in situ denitrification technology based on carbon substrate injections. In advance of the pilot test, detailed characterization of the site was undertaken. The aquifer consisted of complex, discontinuous and interstratified silt, sand and gravel units, similar to other well studied aquifers of glaciofluvial origin, 15-40 m deep. Laboratory and field tests, including a conservative tracer test, a pumping test, a borehole flowmeter test, grain-size analysis of drill cuttings and core material, and permeameter testing performed on core samples, were performed on the most productive depth range (27-40 m), and the results were compared. The velocity profiles derived from the tracer tests served as the basis for comparison with other methods. The spatial variation in K, based on grain-size analysis, using the Hazen method, were poorly correlated with the breakthrough data. Trends in relative hydraulic conductivity (K/K(avg)) from permeameter testing compared somewhat better. However, the trends in transient drawdown with depth, measured in multilevel sampling points, corresponded particularly well with those of solute mass flux. Estimates of absolute K, based on standard pumping test analysis of the multilevel drawdown data, were inversely correlated with the tracer test data. The inverse nature of the correlation was attributed to assumptions in the transient drawdown packages that were inconsistent with the variable diffusivities encountered at the scale of the measurements. Collectively, the data showed that despite a relatively low variability in K within the aquifer under study (within a factor of 3), water and solute mass fluxes were concentrated in discrete intervals that could be targeted for later bioremediation. PMID- 16246460 TI - Airflow dispersion in unsaturated soil. AB - Dispersion data is abundant for water flow in the saturated zone but is lacking for airflow in unsaturated soil. However, for remediation processes such as soil vapour extraction, characterization of airflow dispersion is necessary for improved modelling and prediction capabilities. Accordingly, gas-phase tracer experiments were conducted in five soils ranging from uniform sand to clay at air dried and wetted conditions. The disturbed soils were placed in one-dimensional stainless steel columns, with sulfur hexafluoride used as the inert tracer. The tested interstitial velocities were typical of those present in the vicinity of a soil vapour extraction well, while wetting varied according to the water-holding capacity of the soils. Results gave dispersivities that varied between 0.42 and 2.6 cm, which are typical of values in the literature. In air-dried soils, dispersion was found to increase with the pore size variability of the soil. For wetted soils, particle shape was an important factor at low water contents, while at high water contents, the proportion of macroporous space filled with water was important. The relative importance of diffusion decreased with increasing interstitial velocity and water content and was, in general, found to be minor compared to mechanical mixing across all conditions studied. PMID- 16246461 TI - Follow-up study on contamination rate of expressed breast milk samples and necrotizing enterocolitis in Chinese mothers. PMID- 16246462 TI - Challenges in the epidemiological investigation of the relationships between physical activity, obesity, diabetes, dementia and depression. AB - There are many challenges facing epidemiologists wishing to investigate relationships between physical activity, obesity, diabetes, dementia and depression, all of which are complex fields in their own right. There is a large literature investigating the relationship between diabetes and dementia but less, as yet, on the other exposures and outcomes. In this literature there is a diversity of definitions making rigorous systematic review problematic. There is a need to define hypotheses in this area very clearly and to identify studies that have addressed the specific question. Such exercises have not been carried out to date but would enlighten the research area and point more clearly to questions which remain to be answered. Our own research group has examined the specific question of risk of development of dementia in relation to levels of HbA(1)c, as a marker of glycaemic control and showed that although not related to dementia, it is related to incidence of severe cognitive impairment. PMID- 16246463 TI - Links between cognitive impairment in insulin resistance: an explanatory model. AB - Cognitive function and peripheral glucose regulation both decrease with age. There is a consistent and growing literature reporting memory and other cognitive problems among individuals with diabetes mellitus as well as those with pre diabetes. There are two papers in the literature documenting, relative to matched controls, hippocampal volume reductions among both diabetics and insulin resistant individuals. The mechanism(s) for the cognitive and brain problems associated with impairments in peripheral glucose regulation remain unknown. In this paper, I present a selective review of the literature that builds a case for a theoretical model that could be used to investigate how abnormalities in peripheral glucose regulation may give rise to brain impairments in general, and affect hippocampal integrity in particular. PMID- 16246464 TI - Long-term antioxidant supplementation attenuates oxidative stress markers and cognitive deficits in senescent-accelerated OXYS rats. AB - Oxidative damage of biomolecules increases with age and is postulated to be a major causal factor of various neurodegenerative disorders. Consequently, the concept of neuroprotection by antioxidants has been developed. Recently we have shown that the behaviour of young senescent-accelerated OXYS rats is similar to the behaviour of old Wistar animals. To determine the role of oxidative stress in this phenomenon we investigated age-related changes in protein carbonyls (PrC), lipid peroxides (LP), reduced glutathione (GSH), alpha-tocopherol (TP) and SOD activity in the brain of OXYS and Wistar rats. We also studied the effect of long term supplementation with bilberry extract (2g/kg of diet) and Vitamin E (140 mg/kg of diet) on oxidative stress markers and on learning in passive avoidance test. In both rat strains LP, PrC and TP increased with age and at 24 months PrC was significantly higher (p<0.0001) in OXYS rats. At 3 months GSH was higher and SOD activity was lower in OXYS rats than in Wistar rats. SOD activity decreased with age in OXYS whereas increased in Wistar rats. Cognitive impairments in OXYS rats were manifested earlier than significant differences in the level of brain oxidative stress markers between two strains. By contrast, differences in antioxidant systems of Wistar and OXYS rats were registered at 3 months. Antioxidants attenuated cognitive deficits in OXYS rats, providing evidence for therapeutic role of antioxidants. Nevertheless, the exact mechanisms of neuroprotective effects of antioxidants in vivo and the real impact of oxidative stress on the development of cognitive impairments in OXYS rats still needs to be further investigated. PMID- 16246465 TI - Motor-response generation as a source of aging-related behavioural slowing in choice-reaction tasks. AB - OBJECTIVE: To analyze the effects of stimulus-response (SR) processing modes on different central stages of sensorimotor processing in order to evaluate their contribution to aging-related behavioural slowing. METHODS: Components of stimulus- and response-related potentials (ERPs/RRPs) and lateralized readiness potentials (LRPs) were analyzed in two groups of young (mean 22.5 years) and older adults (mean 58.3 years) during an auditory and a visual four-choice reaction task. RESULTS: (1) Reaction time (RT) depended on the SR type, indicating SR-specific differences in processing, which did not vary with age. (2) For each SR type, the RT increased with age. RT slowing was not accompanied by significant delays in early stimulus processing (as reflected by P1 and N1 latencies) nor in response selection (as reflected by the onset of stimulus locked LRP), but resulted from a prolongation of contralateral motor activity during motor response execution indexed by earlier, longer durated and larger motor-related potentials in older adults. These aging effects were observed for each SR type. CONCLUSION: In a four-choice-reaction task, (1) task complexity rather than differences in cognitive strategy or activation patterns subserving SR-specific processing leads to response slowing with aging and (2) the most plausible contributor to this slowing is the cortical response generation system. PMID- 16246466 TI - Effect of delivery parameters on immunization to ovalbumin following intracutaneous administration by a coated microneedle array patch system. AB - Immunization to the model antigen ovalbumin was investigated using a novel intracutaneous delivery system consisting of antigen-coated microneedle arrays. The influence of the following parameters on the resulting immune responses was investigated: depth of vaccine delivery, dose of vaccine delivered, density of microneedles on the array, and area of application. The immune response was found to be dose dependent, and mostly independent of depth of delivery, density of microneedles, or area of application. Our studies show that the shortest, most tolerable microneedle arrays can be used for achieving consistent and high antibody titers. Overall, the microneedle array proves to be a very versatile delivery technology, allowing easy and reproducible antigen delivery to skin for efficient vaccination without the use of a needle and syringe. PMID- 16246467 TI - Photopolymerized hydrogel carriers for live vaccine ballistic delivery. AB - Photopolymerized poly(ethylene glycol) (PEG)-crosslinked hydrogels were assessed for their ability to serve as a payload vehicle to deliver a viable bacterial vaccine (Brucella abortus strain RB51 (RB51) to bison in Yellowstone National Park) ballistically using thermoplastic degradable Biobullets. PEG modified with degradable glycolide or lactide oligomers capped with photopolymerizable methacrylate groups served to crosslink the hydrogel vaccine carrier inside commercial hydroxypropylcellulose Biobullets. Release of 1 microm diameter model fluorescent particles from hydrogels followed known degradation trends for glycolide- and lactide-modified PEG hydrogels. All particles were released from PEG-co-glycolide hydrogels after approximately 10 days and PEG-co-lactide hydrogels after approximately 45 days following gel degradation. Minimal particle release was observed from pure PEG dimethacrylate hydrogels over 40 days. P. aeruginosa (strain PAO1) and RB51 live vaccines exhibit excellent viability following exposure to photopolymerization encapsulation within these gel matrices. Hydrogels photopolymerized into the payload chamber of Biobullets exhibit similar ballistic properties to commercially available Biobullets and penetrate and remain intact when fired intramuscularly into live elk for release of their gel payload in the host. PMID- 16246468 TI - Relationship of adsorption mechanism of antigens by aluminum-containing adjuvants to in vitro elution in interstitial fluid. AB - The objective of this research was to determine how the mechanism by which antigens adsorb to aluminum-containing adjuvants affects the elution upon exposure to interstitial fluid. Antigens (alpha lactalbumin, bovine serum albumin, lysozyme and myoglobin) that adsorb to aluminum-containing adjuvants principally by electrostatic attraction were found to elute readily in vitro when exposed to interstitial fluid. Phosphorylated antigens (alpha casein, hepatitis B surface antigen and phosphorylated bovine serum albumin) that adsorb to aluminum containing adjuvants principally by ligand exchange exhibit little if any elution during 12-24 h in vitro exposure to interstitial fluid. Dephosphorylated alpha casein, which contains less than two phosphate groups, was less strongly adsorbed by ligand exchange in comparison to alpha casein, which contains eight phosphate groups. Dephosphorylated alpha casein was completely eluted when exposed to interstitial fluid. The results of this study lead to the generalization that antigens that adsorb to aluminum-containing adjuvants by electrostatic attraction are more likely to elute upon intramuscular or subcutaneous administration than antigens that adsorb by ligand exchange. PMID- 16246469 TI - Hexa-acylation and KDO(2)-glycosylation determine the specific immunostimulatory activity of Neisseria meningitidis lipid A for human monocyte derived dendritic cells. AB - To better understand immune modulation by endotoxin and facilitate the development of novel vaccine adjuvants, the structural requirements of Neisseria meningitidis lipopoly(oligo)saccharide (LOS) for activation of human monocyte derived dendritic cell (MDDC) was determined. Highly purified LOS from wild type and genetically-defined mutants of N. meningitidis serogroup B were used. Unglycosylated or penta-acylated meningococcal KDO(2)-lipid A failed to induce human MDDC maturation and activation. However, both wild type meningococcal LOS and KDO(2)-lipid A, significantly up-regulated CD80, CD83 and CD86 and released significantly higher amounts of IL-12p70, IL-6, IL-10, TNFalpha, MCP-1, IP-10 and RANTES. Further, DCs stimulated with wild type or KDO(2)-lipid A but not meningococcal lipid A or penta-acylated KDO(2)-lipid A stimulated naive allogeneic CD4+ T cells to secrete enhanced levels of IFN-gamma, relative to T cells primed with immature DCs. In contrast to Escherichia coli LPS, IL-5 production was enhanced or maintained in CD4+ T-cells stimulated with MDDC exposed to wild-type meningococcal LOS and KDO(2)-lipid A. These data suggest that KDO linked to a fully acylated meningococcal lipid A is required for meningococcal endotoxin's immunostimulatory activity of human MDDC via TLR4/MD-2 and that different endotoxin structures influence Th responses mediated by MDDC. PMID- 16246470 TI - An evaluation of the effectiveness of an educational programme promoting the introduction of routine antenatal enquiry for domestic violence. AB - OBJECTIVE: a feasibility study to evaluate the effect of an educational programme on midwives' knowledge, skills, attitudes and implementation of routine antenatal enquiry for domestic violence. DESIGN: pre-, post- and follow-up survey. SETTING: an acute Trust within the South West of England. PARTICIPANTS: 79 of the 82 community midwives (96%) working in the Trust participated in the training programme, with 70 (85%) participating at all three stages of the research. MEASUREMENTS: participating community midwives completed a 38-item questionnaire at three points during the study: before the educational programme to provide base-line data, post-test immediately after the programme, and at 6 months follow up. The questionnaire was divided into the following categories: views of professional education, knowledge of domestic violence, attitudes to domestic violence, efficacy beliefs and issues of practice development. The aim of the study was to identify any differences between pre- and post-implementation test data in relation to all the areas identified. Repeated multivariate analysis of variance was used to examine changes between pre-, post- and follow-up measures of knowledge, attitudes and efficacy. Hierarchical regression was used to identify potential influences on post-training disclosure rates using pre-, post- and follow-up measures as predictors. FINDINGS: the programme was positively received by participants, particularly in relation to an increased awareness and confidence in dealing with domestic violence. It was also associated with improvements in knowledge, attitudes and efficacy at post-test. These changes declined but remained above pre-test levels at 6 months follow-up. Levels of current and previous experiences of abuse obtained by midwives were predicted by past experience of dealing with the issue and efficacy scores immediately after and at 6 months after programme delivery. Rates of enquiry after programme introduction were lower than anticipated, with midwives routinely asking only 50% of the time. However, the key barrier identified was the presence of a male partner. IMPLICATIONS FOR PRACTICE: the effect of routine enquiry for domestic violence on midwifery role development needs further exploration before universal introduction. Seeing women alone at least once during a pregnancy would clearly increase opportunities for directly asking about violence and allowing safe disclosure. Where enquiry is introduced, midwives should be given access to validated educational programmes and structured ongoing support if enquiry is to be sustained over time. Although further evaluations are necessary, it may be advisable to focus on skills-based programmes that increase midwives' confidence and prioritise support and safety aspects for midwives and women during enquiry about domestic violence. PMID- 16246471 TI - Fatigue resistance analysis of tibial baseplate in total knee prosthesis--an in vitro biomechanical study. AB - BACKGROUND: Tibial baseplates were occasionally reported with clinical fatigue failures. This study postulated that tibial baseplate of a specific mobile bearing design with a uniform thickness across the baseplate offers more fatigue resistance than the fixed-bearing design. Tibial baseplates of a fixed bearing and a mobile bearing design were fatigue-tested in vitro to study their fatigue resistance. METHODS: Five samples of each design were tested under a sinusoidal loading between 90 N and 900 N at 30 Hz till failure or 10 million cycles. Experimental setup followed a standard published test method. Scanning electron microscope was used for inspecting the fracture surface of the failed baseplate. FINDINGS: Two baseplates of fixed bearing design failed before 10 million cycles. Fatigue crack advancement marks were visible on the fractured surface of the failed samples. The fractured cross-section showed that the failure started near the end of the fin, it was likely due to the stress concentration as stress singularity existed at a point of sudden geometrical change. Five mobile bearing baseplates passed the test. Design of the tibial baseplate without fin structure and with a uniform thickness across the whole baseplate could help reducing the incidence of fatigue failure. INTERPRETATION: The prosthesis survival rate was influenced by the long-term integrity of the metallic part of the prostheses such as the tibial baseplate. This study revealed that the tibial baseplate of a mobile bearing design with a uniform thickness provided better fatigue resistance than fixed bearing one. Standardized fatigue screening of the tibial baseplate was considered important in designing knee prostheses. PMID- 16246472 TI - Effects of nitrogen with and without acidified sulphur on an ectomycorrhizal community in a Sitka spruce (Picea sitchensis Bong. Carr) forest. AB - This preliminary study investigated the effects of enhanced nitrogen (NH4NO3 at 48 kg ha(-1) y(-1)), sulphur (Na2SO4 at 50 kg ha(-1) y(-1)), acidified nitrogen and sulphur (H2SO4 + NH4NO3) at pre-stated doses (pH 2.5), and acidified nitrogen and sulphur deposition at double these doses on the ectomycorrhizal community associated with a 13-year-old Sitka spruce (Picea sitchensis) forest. Sulphur deposition had little impact on below ground ectomycorrhizal diversity, but stimulated sporocarp production. Nitrogen inputs increased below ground colonisation compared to acidified nitrogen and sulphur, largely due to an increase in Tylospora fibrillosa colonisation. Sporocarp production and ectomycorrhizal root colonisation by Lactarius rufus were reduced in the nitrogen treated plots. These observations suggest that nitrogen deposition to a young plantation may suppress ectomycorrhizal fungi producing large sporocarps. It is proposed that enhanced nitrogen deposition increases ectomycorrhizal nitrogen assimilation, consuming more carbon and leaving less for extrametrical mycelium and sporocarp development. PMID- 16246473 TI - Degradation of polycyclic aromatic hydrocarbons (PAHs) in an aged coal tar contaminated soil under in-vessel composting conditions. AB - In-vessel composting of polycyclic aromatic hydrocarbons (PAHs) present in contaminated soil from a manufactured gas plant site was investigated over 98 days using laboratory-scale in-vessel composting reactors. The composting reactors were operated at 18 different operational conditions using a 3-factor factorial design with three temperatures (T, 38 degrees C, 55 degrees C and 70 degrees C), four soil to green waste ratios (S:GW, 0.6:1, 0.7:1, 0.8:1 and 0.9:1 on a dry weight basis) and three moisture contents (MC, 40%, 60% and 80%). PAH losses followed first order kinetics reaching 0.015 day(-1) at optimal operational conditions. A factor analysis of the 18 different operational conditions under investigation indicated that the optimal operational conditions for degradation of PAHs occurred at MC 60%, S:GW 0.8:1 and T 38 degrees C. Thus, it is recommended to maintain operational conditions during in-vessel composting of PAH-solid waste close to these values. PMID- 16246474 TI - Sequential extraction combined with isotope analysis as a tool for the investigation of lead mobilisation in soils: application to organic-rich soils in an upland catchment in Scotland. AB - Sequential extraction (modified BCR procedure) combined with isotope analysis has been investigated as a tool for assessing mobilisation of lead into streams at an upland catchment in NE Scotland. The maximum lead concentrations (up to 110 mg kg(-1) in air-dried soil) occurred not at the surface but at about 10 cm depth. The lowest (206)Pb/(207)Pb ratios in any profile occurred, with one exception, at 2.5-5 cm depth. In the one exception, closest to the only road in the area, significantly lower (206)Pb/(207)Pb ratios in the surface soil together with much increased chloride concentrations (in comparison to other surface waters) indicated the possible mobilisation of roadside lead and transfer to the stream. The (206)Pb/(207)Pb ratios in extractable fractions tended at depth towards the ratio measured in the residual phase but the ratios in the oxidizable fraction increased to a value higher than that of the residual phase. PMID- 16246475 TI - Remobilization of metals from slag and polluted sediments (case study: the canal of the Deule River, northern France). AB - The anthropogenic impact on the environment in the last century has proven to be very negative due to the fast development of industry. A typical example is the Deule River in northern France, one of the most polluted sites in this region. The concentrations of Pb, Cd, Zn and Cu in river sediments are 300, 800, 50 and 15 times higher, respectively, than the background values. The present study was undertaken to evaluate the capacity of already polluted sediments to capture metals released from industrial wastes (slag). As it were, in spite of the high metal pollution level, sediments have still shown the ability to adsorb metals released from slag under the conditions provided. Their efficiency in "cleaning up" some of the metals (e.g. Pb) seems to be additionally enhanced in anoxic conditions. This study provided some additional information on the importance of sediments as a pollutant sink. PMID- 16246476 TI - Effects of polycyclic aromatic hydrocarbons on germination and subsequent growth of grasses and legumes in freshly contaminated soil and soil with aged PAHs residues. AB - The relevance of germination trials for screening plants that may have potential for use in the phytoremediation of PAH contaminated land was evaluated. The germination and subsequent growth of 7 grass and legume species were evaluated in soil spiked with a pure PAH mixture or coal tar and soil from a former coking plant heavily contaminated with aged PAHs. None of these treatments adversely affected germination of the plants. However, apart from Lolium perenne all species exhibited reduced growth in the coking plant soil after 12 weeks growth when compared to the untreated soil. In the coal tar spiked soil 4 out of the 7 species showed reduced growth, as did 3 out of the 7 in the soil spiked with a mixture of 7 PAHs. Therefore, germination studies alone would not predict the success of subsequent growth of the species tested in the ranges of soil PAH levels studied. PMID- 16246477 TI - Antimony mobility in Japanese agricultural soils and the factors affecting antimony sorption behavior. AB - The mobility of antimony (Sb) in Japanese agricultural soils was studied by radiotracer experiments using 124Sb tracer. The soil-solution distribution coefficients (Kd) of Sb were measured for 110 soil samples. These Kds ranged from 1 to 2065 L kg(-1); the geometric mean was 62 L kg(-1) excluding one extremely high value, 2065 L kg(-1). Experimental measurement of Kd showed a decrease with both increasing pH and increasing phosphate concentration. The latter suggested that one aspect of the Sb sorption phenomena in Japanese soil was influenced by specific adsorption of anions such as phosphate. However, other aspects could not be explained by this specific adsorption mechanism, because only 20-40% of soil sorbed Sb could be extracted by phosphate solution. PMID- 16246478 TI - Effects of triadimefon on extracellular dopamine, DOPAC, HVA and 5-HIAA in adult rat striatum. AB - Triadimefon has been shown to inhibit monoamine uptake, bind to the dopamine (DA) transporter, and stimulate dopamine efflux in rat brain tissue, in vitro. To determine whether these changes also occur in the intact animal and to study the reversibility of the effects observed, we used in vivo microdialysis to determine changes in the concentrations of DA, dihydroxyphenylacetic acid (DOPAC), homovanillic acid (HVA) and 5-hydroxyindolacetic acid (5-HIAA) in the striatal dialysates from free moving adult rats after exposure to triadimefon 50, 100 and 200mg/kg, i.p. Triadimefon induced a gradual dose- and time-dependent accumulation of extracellular DA accompanied by a small increase in the HVA and 5 HIAA concentrations. These changes were still present 24h after treatment in the group treated with 200mg/kg and had vanished 48 h after treatment. In contrast to the DA efflux induced by S(+)-amphetamine (2mg/kg, i.p.), that induced by triadimefon was totally inhibited by the infusion of 10(-5)M tetrodotoxin (TTX), a voltage-gated Na(+) channel blocker, thus showing that the increase in extracellular DA induced by triadimefon was an action potential-dependent mechanism. GBR 12909 (10mg/kg, i.p.), a dopamine uptake inhibitor, induced a gradual increase in striatal dopamine similar to that induced by triadimefon, whereas the effects on the acid metabolites were not exactly the same. The present results indicate that triadimefon acts in vivo as a DA transporter inhibitor and could also act on the serotoninergic system. PMID- 16246479 TI - Tetrandrine-induced apoptosis in rat primary hepatocytes is initiated from mitochondria: caspases and endonuclease G (Endo G) pathway. AB - Tetrandrine, a bisbenylisoquinoline alkaloid isolated from the dried root of Stephenia tetrandra (S Moore), possesses a remarkable pharmacological profile. However, the mechanisms of tetrandrine hepatotoxicity remain to be elucidated. In this study, we first proved apoptosis and mitochondrial dysfunction induced by tetrandrine in Sprague-Dawley rat liver in vivo. By further assuming apoptosis as an important mechanism in tetrandrine-induced hepatotoxicity, we focused on mitochondria-initiated apoptosis in primary hepatocytes isolated from Sprague Dawley male rats. Tetrandrine treatment led to significant release of cytochrome c and downregulation of Bcl-X(L) accompanied by caspase 3 activation, and ultimately, DNA fragmentation. Caspase 3 activation was markedly inhibited by cyclosporin A (CsA) and Ac-DEVD-CHO. Furthermore, Endo G, a caspase-independent apoptotic protein, was detected for its expression and DNase activity. CsA blocked the release both of Endo G and cytochrome c significantly. Additionally, the generation of reactive oxygen species (ROS) increased in a time-dependent manner corresponding with a fall in intracellular GSH content after 10 microM tetrandrine treatment in 4h. Tetrandrine also induced mitochondrial dysfunction indicated by transition of mitochondrial transmembrane potential and decrease of intracellular ATP level. The findings indicated that the caspase-dependent mitochondrial apoptosis pathway was primarily involved in tetrandrine-induced apoptosis in rat primary hepatocytes. In addition, a caspase-independent pathway indicated by Endo G also contributed to apoptosis caused by tetrandrine. Meanwhile, ROS was proved an important inducer in this apoptosis process. PMID- 16246480 TI - Outcome of direct restorations placed within the general dental services in England and Wales (Part 3): variation by dentist factors. AB - AIM: It is the aim of this paper to investigate the survival of direct-placement restorations provided within the General Dental Services in England and Wales, in relation to the dentist factors which may affect this. METHODS: For this work, survival of a restoration was considered to be the time between the date of completion of the course of treatment in which it was placed and the date of acceptance of the course of treatment when the next tooth-specific treatment was carried out on the same tooth. A modified version of Kaplan-Meier statistical methodology was used to plot survival curves for restorations placed by different subgroups of dentists. RESULTS: The age and experience of the dentist who placed the original restoration were found to be related to restoration survival, with older dentists having shorter intervals from placement of restorations to re intervention (P<0.001). Country of qualification seems not to be relevant within Europe in terms of restoration survival, but dentists who qualified outside Europe achieve different restoration survival times for the restorations that they place. Dentist's gender has no relationship with time from restoration to re intervention. CONCLUSIONS: Dentist factors such as age, country of qualification, and employment status, but not the gender of the dentist, have been found to influence the survival of directly placed restorations. PMID- 16246481 TI - The management of endometrial hyperplasia: an evaluation of current practice. AB - OBJECTIVE: To identify current management practices and evaluate subsequent outcomes of treatment for women diagnosed with endometrial hyperplasia. STUDY DESIGN: All women with a histological diagnosis of endometrial hyperplasia at the Birmingham Women's Hospital were identified between October 1998 and September 2000. A retrospective case note review was performed for each woman using a standardised data abstraction sheet. Baseline characteristics including clinical presentation and treatment strategy were obtained. Results of subsequent endometrial tissue examinations were used to assess histological response to treatment and the need and indication for hysterectomy was used to assess clinical response. RESULTS: There were 351 women diagnosed with endometrial hyperplasia during the study period of which 84% presented with symptoms of abnormal uterine bleeding and 54% were postmenopausal. Complex endometrial hyperplasia was the most common diagnosis accounting for 60% of all cases. Eighty percent of women with atypical endometrial hyperplasia were treated by hysterectomy compared with 30% without evidence of cytological atypia (relative hysterectomy rate of 2.6, 95% CI 2.0-3.3). Hysterectomy was avoided in 138/172 (80%, 95% CI 74-86%) women managed conservatively during the study period. Overall 35/108 (36%, 95% CI 27-46%) of women managed conservatively had persistent or progressive disease identified (mean follow up 36 months). 20/143 (14%) women initially diagnosed with endometrial hyperplasia who subsequently underwent hysterectomy were found to have endometrial cancer, the majority of whom had been diagnosed with atypical disease (14/20, 70%). CONCLUSION(S): The majority of women with atypical endometrial hyperplasia were managed by hysterectomy and the substantial risk of diagnostic under-call supports this approach to treatment. In contrast, there is no consensus regarding the initial management of women with endometrial hyperplasia without cytological atypia. PMID- 16246482 TI - A new predictive scoring system including shock index for unruptured tubal pregnancy patients. AB - OBJECTIVE: Shock index (SI) and predictive score grading system including it for predicting medical treatment failure of tubal pregnancies have been studied. STUDY DESIGN: Eighty-eight patients were diagnosed as nonruptured tubal pregnancies. Shock index was calculated as the ratio of heart rate to systolic arterial pressure. A predictive score was used based on four parameters including initial level of beta-human chorionic gonadotropin (betahCG), aspect of the image on ultrasound, size of the ectopic mass and shock index value at admission. RESULTS: Forty patients have undergone to surgery because of tubal gestational sac size > or =4 cm and/or presence of fetal heart activity. Nineteen patients were managed expectantly. Twenty-four patients received single dose methotrexate (MTX) and five patients received second dose MTX. Success rate for single dose MTX therapy was 72% (21/29). The cut-off shock index value for tubal rupture was 0.77 with 89% sensitivity and 61% specifity. CONCLUSION: In this study, we demonstrated that tubal pregnancy patients who were managed with nonsurgical measures at admission and who had SI values lower than 0.77 and predictive score grades greater and equal to 6.5 did not experience tubal rupture and did not need surgical intervention during nonsurgical management. PMID- 16246483 TI - Fibrosarcoma: a rare ovarian tumor. PMID- 16246484 TI - Multithermal titration calorimetry: a rapid method to determine binding heat capacities. AB - Herein a new method that allows binding DeltaCp to be determined with a single experiment is presented. Multithermal titration calorimetry (MTC) is a simple extension of isothermal titration calorimetry (ITC) that explicitly takes into account the thermal dependences of DeltaH and the binding constant. Experimentally, this is accomplished by performing a single stepwise titration with ITC equipment, allowing temperature re-adjustments of the system at intermediate states of the titration process. Thus, from the resulting multitherm, DeltaCp can also be determined. The experimental feasibility of MTC was tested by using the well-characterized lysozyme-chitotriose complex as a model system. PMID- 16246485 TI - Alpha-benzene hexachloride exerts hormesis in preneoplastic lesion formation of rat hepatocarcinogenesis with the possible role for hepatic detoxifying enzymes. AB - Recently there has been a shift in the prevailing paradigm regarding the dose dependence of carcinogen action with increasing acceptance of hormesis phenomenon, although underlying mechanisms remain to be established. To ascertain whether alpha-benzene hexachloride (alpha-BHC) might act by hormesis, rats were initiated with diethylnitrosamine and then alpha-BHC ranging from 0.01 to 500 ppm was administered in the diet for 10 weeks. The highest concentration of alpha-BHC significantly increased the number and area of glutathione S-transferase placental form (GST-P) positive foci, preneoplastic lesions in the liver, but its low dose, 0.05 ppm, caused significant reduction, showing a J-shape dose-response curve. The proliferating cell nuclear antigen positive index for GST-P positive foci in the low dose-treated group was significantly reduced. The dose response curves of CYP450 content, NADPH-P450 reductase activity and 8 hydroxydeoxyguanosine formation revealed the same pattern as GST-P positive foci data. The response curves of CYP2B1 and 3A2 in their activities, protein and mRNA expression showed a threshold but CYP2C11 activity exhibited an inverted J-shape. These results might suggest the possibility of hormesis of alpha-BHC at early stages of rat hepatocarcinogenesis. The possible mechanism involves induction of detoxifying enzymes at low dose, influencing free radical production and oxidative stress, and consequently pathological change in the liver. PMID- 16246486 TI - DAPK promotor methylation is an early event in colorectal carcinogenesis. AB - Death-associated protein kinase (DAPK) is frequently inactivated by promotor hypermethylation in various human cancers. At present, little is known about the significance of DAPK inactivation in colorectal carcinogenesis. We therefore, investigated macrodissected samples of 22 formalin-fixed and paraffin-embedded T1 carcinomas showing normal colon mucosa, intraepithelial neoplasia and carcinoma tissue on the same slice. Dissected carcinoma areas showed a higher frequency of DAPK promotor methylation (81.2%) compared to intraepithelial neoplasia (68.2%). Colon mucosa adjacent to intraepithelial neoplasia or carcinoma showed DAPK promotor methylation in only two of eight cases (25%). We suggest that DAPK promotor hypermethylation may play an important role in the early steps of tumor progression in colorectal carcinoma. PMID- 16246487 TI - Disruption of the Fanconi anemia/BRCA pathway in sporadic cancer. AB - An increasing number of studies provide evidences linking disruption of Fanconi anemia/BRCA cascade with sporadic cancers. Given that this pathway plays essential roles in response to the DNA interstrand cross-links, these cancers are expected to be chemosensitive to cross-link based therapy. In the present mini review we expand the spectrum of possibilities for FA/BRCA disruption and review some works describing functional upstream and downstream events linking disruption of the FA/BRCA pathway to sporadic cancer. This may involve but not limited to epigenetic silencing of the FA-core complex or BRCA1/2, mutations of one or several FA-BRCA genes or modification of encoded products. All this may serve as a platform for occurrence, development and treatment of sporadic cancers and therefore deserves to be in the focus of new research directions. PMID- 16246488 TI - Genotoxic damage in female residents exposed to environmental air pollution in Shenyang city, China. AB - Air pollution has been suggested to cause genetic damage from investigations of many biological markers that measure cytogenetic damage in humans. Here, we evaluated the genotoxic effects of ambient air pollution by investigating the extent of cytogenetic damage in human blood lymphocytes from rural and industrial female residents of Shenyang city, China, using micronuclei assays and polymorphic analyses of metabolic enzyme and DNA repair genes. After adjustment for potential confounding factors including DNA polymorphisms, industrial female residents were found to have a higher micronuclei frequency. These results provide evidence that micronuclei assays are a sensitive indicator to air pollution-induced genotoxic effects in humans. PMID- 16246489 TI - Combination vaccine of dendritic cells (DCs) and T cells effectively suppressed preestablished malignant melanoma in mice. AB - The study aims at establishing a novel vaccine procedure, using bone marrow derived DCs that have ingested apoptotic B16 melanoma (DCs(+)), alone or in combination with splenic T lymphocytes from a syngenic donor. Co-immunization with DCs(+) and T cells showed the highest antitumor potential against preestablished B16 tumor in mice, in which CTL and NK cytotoxicities were drastically elevated, while either DCs(+) alone, naive DCs (DCs(-)) alone, or a mixture of DCs(-) and T cells induced less significant therapeutic outcomes. Use of extracellular matrix proteins elevated antitumor activity of DC(-)/T cell vaccine. Compared with the CD8(+) cells, the CD4(+)T cells more remarkably improved the efficacy of DC-based immunotherapy. The present system may be a feasible therapeutic modality to eradicate malignancies including melanoma. PMID- 16246490 TI - Treatment of organophosphate-contaminated wastewater by acidic hydrolysis and precipitation. AB - Wastewater from commercial manufacture of organophosphate (OP) pesticide of O methyl-O-(2-isopropyl salicylate) thiophosphorusyl amide contains large amounts of organophosphate, organic sulfides, COD and NH3-N. A treatment process including acidic hydrolysis and precipitation was explored in this paper. The effects of initial pH values, hydrolysis temperature and hydrolysis time on the removal of pollutants were investigated. The experimental results showed that, in the hydrolysis, with the decreasing initial pH value, the removal of OP, sulfide and COD rose whereas the NH3-N removal declined. Increased hydrolysis temperature and time favored the removal of all the pollutants. 15.0 wt% Ca(OH)2 was sufficient for the removal of inorganic phosphorus and residual NH3-N in the precipitation step. With this chemical process, the removal of total phosphorus (TP), organophosphate, sulfides and NH3-N exceeded 90%, and the removal of COD was about 60%. The BOD5/COD value was greatly improved from 0.05 to 0.3, and the bio-degradability of the wastewater was greatly improved. The NH3 generated in the process was absorbed with water and might be reused in the manufacture of the pesticide. The results showed that this comprehensive process is effective for the treatment of this typical unbio-degradable pesticide wastewater. PMID- 16246491 TI - IEC 61511 and the capital project process--a protective management system approach. AB - This year, the process industry has reached an important milestone in process safety-the acceptance of an internationally recognized standard for safety instrumented systems (SIS). This standard, IEC 61511, documents good engineering practice for the assessment, design, operation, maintenance, and management of SISs. The foundation of the standard is established by several requirements in Part 1, Clauses 5-7, which cover the development of a management system aimed at ensuring that functional safety is achieved. The management system includes a quality assurance process for the entire SIS lifecycle, requiring the development of procedures, identification of resources and acquisition of tools. For maximum benefit, the deliverables and quality control checks required by the standard should be integrated into the capital project process, addressing safety, environmental, plant productivity, and asset protection. Industry has become inundated with a multitude of programs focusing on safety, quality, and cost performance. This paper introduces a protective management system, which builds upon the work process identified in IEC 61511. Typical capital project phases are integrated with the management system to yield one comprehensive program to efficiently manage process risk. Finally, the paper highlights areas where internal practices or guidelines should be developed to improve program performance and cost effectiveness. PMID- 16246492 TI - P50 sensory gating deficit in children with centrotemporal spikes and sharp waves in the EEG. AB - Sensory gating refers to the ability of the brain to inhibit irrelevant sensory input. In several studies, a pathogenic role of the CHRNA7 gene and the CHRNA7 like gene, respectively, is suggested. In linkage analysis concerning familial centrotemporal spikes and sharp waves (CTS) and benign rolandic epilepsy, evidence for linkage was found to a region on chromosome 15q14, close to the alpha-7 subunit gene of the neuronal nicotinic acetylcholine receptor (CHRNA7). According to these findings, P50 evoked potentials to paired click stimuli were studied in 13 children with CTS in the EEG to determine whether they had normal sensory gating. The control group consisted of 13 healthy probands matched for gender and age. Children with CTS showed a significant sensory gating deficit (p=0.001). These results (1) suggest an inhibitory deficit in early pre-attentive auditory sensory processing in children with CTS and (2) confirm the assumption of a cholinergic pathology in CTS. PMID- 16246493 TI - A content analysis of personal strivings: associations with substance use. AB - This study examined relations between personal strivings and frequency of alcohol, marijuana, and any drug use among college students. Personal strivings are ongoing goals that an individual is characteristically trying to achieve through their behavior. Participants generated lists of personal strivings following standard instructions and then completed an assessment of alcohol and marijuana use and days used any drug (aside from tobacco or medication as prescribed). Personal strivings were coded into content categories by trained raters using a coding manual. Four content categories were examined for this study: achievement, affect regulation, self-presentation, and interpersonal. A series of t-tests revealed that participants' achievement strivings were unrelated to substance use. In contrast, participants endorsing more affect regulation goals used all drugs more frequently. Self-presentation goals were positively associated with alcohol use but unrelated to marijuana or other drug use. Finally, interpersonal goals were associated with higher rates of alcohol use, lower rates of marijuana use, and not related to total use days. PMID- 16246494 TI - Eleven-year follow-up results in the delay of breast irradiation after conservative breast surgery in node-negative breast cancer patients. AB - PURPOSE: This retrospective review was conducted to determine if delay in the start of radiotherapy after conservative breast surgery had any detrimental effect on local recurrence or disease-free survival in node-negative breast cancer patients. METHODS AND MATERIALS: A total of 568 patients with T1 and T2, N0 breast cancer were treated with breast-conserving surgery and breast irradiation, without adjuvant systemic therapy, between January 1, 1985 and December 31, 1992 at the London Regional Cancer Centre. The time intervals from definitive breast surgery to breast irradiation used for analysis were 0 to 8 weeks (201 patients), greater than 8 to 12 weeks (235 patients), greater than 12 to 16 weeks (91 patients), and greater than 16 weeks (41 patients). Kaplan-Meier estimates of time to local-recurrence and disease-free survival rates were calculated. RESULTS: Median follow-up was 11.2 years. Patients in all 4 time intervals were similar in terms of age and pathologic features. No statistically significant difference was seen between the 4 groups in local recurrence or disease-free survival with surgery radiotherapy interval (p = 0.521 and p = 0.222, respectively). The overall local-recurrence rate at 5 and 10 years was 4.6% and 11.3%, respectively. The overall disease-free survival at 5 and 10 years was 79.6% and 67.0%, respectively. CONCLUSION: This retrospective study suggests that delay in the start of breast irradiation of up to 16 weeks from definitive surgery does not increase the risk of recurrence in node-negative breast cancer patients. The certainty of these results is limited by the retrospective nature of this analysis. PMID- 16246495 TI - Accelerated partial breast irradiation: an analysis of variables associated with late toxicity and long-term cosmetic outcome after high-dose-rate interstitial brachytherapy. AB - PURPOSE: To perform a detailed analysis of variables associated with late tissue effects of high-dose-rate (HDR) interstitial brachytherapy accelerated partial breast irradiation (APBI) in a large cohort of patients with prolonged follow-up. METHODS AND MATERIALS: Beginning in 1995, 75 women with Stage I/II breast cancer were enrolled in identical institutional trials evaluating APBI as monotherapy after lumpectomy. Patients eligible included those with T1-2, N0-1 (or=2 were scheduled to receive the tetracaine gel. A questionnaire evaluating the effect of the gel was given to all subjects. RESULTS: In 38 patients (79.2%), a reduction in oral cavity pain was reported. Thirty-four patients (82.9%) reported no side effect. Seventy-one percent of patients had no difficulties in gel application. Unpleasant taste of the gel and interference with food taste were noticed in 5 (12%) and 16 patients (39%), respectively. Planned RT course was interrupted less frequently in patients who reported benefit from gel application than in patients who did not (p=0.014). None of the patients who experienced pain relief needed a nasogastric tube, opposite to the patients who did not report any benefit from gel application (p=0.001). CONCLUSION: Tetracaine oral gel administration seemed feasible and safe while reducing RT-induced mucositis-related oral pain in a sizeable proportion of treated head-and-neck cancer patients. A trial designed to compare efficacy of this gel vs. standard treatment is warranted. PMID- 16246499 TI - Antifeedant activity of Momordica dioica fruit pulp extracts on Spodoptera litura. AB - The hexane extract and ethyl acetate soluble fraction of methanolic extract of the fruit pulp of Momordica dioica exhibited moderate and concentration dependent antifeedant activity against Spodoptera litura. PMID- 16246500 TI - Effects of Erigeron breviscapus ethanol extract on neuronal oxidative injury induced by superoxide radical. AB - Antioxidant activity of the ethanol extract of Erigeron breviscapus (EEEB) were studied by using neuron oxidative injury model induced by superoxide radical. EEEB at the dose of 10-140 mug/ml reduced significantly the lipid peroxidation levels and lactic dehydrogenase (LDH) release from neuron exposed to superoxide radical (generated by xanthine and xanthine oxidase). These results suggest that neuroprotective actions of EEEB may be due to its antioxidant or radical scavenging activity. PMID- 16246501 TI - Antioxidative activity of 6a-hydroxypterocarpan from the root of Sophora chrysophylla. AB - The antioxidative activity of 6a-hydroxypterocarpan isolated from the root of Sophora chrysophylla has been investigated in vitro on lipid peroxidation by interaction of heamoglobin and hydrogen peroxide. PMID- 16246502 TI - Myogenic stem cells: regeneration and cell therapy in human skeletal muscle. AB - Human skeletal muscle has been considered as an ideal target for cell-mediated therapy. However, the positive results obtained in dystrophic animal models using the resident precursor satellite cell population have been followed by discouraging evidences obtained in the clinical trials involving Duchenne muscular dystrophy patients. This text reviews the recent advances that many groups have achieved to identify from the stem cell compartment putative candidates for cell therapy. We focused our attention on stem cells with myogenic potential which might be able to improve transplantation efficiency and therefore could be used as a therapeutic tool for neuromuscular diseases. PMID- 16246503 TI - Lateralized visual behavior in bottlenose dolphins (Tursiops truncatus) performing audio-visual tasks: the right visual field advantage. AB - Analyzing cerebral asymmetries in various species helps in understanding brain organization. The left and right sides of the brain (lateralization) are involved in different cognitive and sensory functions. This study focuses on dolphin visual lateralization as expressed by spontaneous eye preference when performing a complex cognitive task; we examine lateralization when processing different visual stimuli displayed on an underwater touch-screen (two-dimensional figures, three-dimensional figures and dolphin/human video sequences). Three female bottlenose dolphins (Tursiops truncatus) were submitted to a 2-, 3- or 4-, choice visual/auditory discrimination problem, without any food reward: the subjects had to correctly match visual and acoustic stimuli together. In order to visualize and to touch the underwater target, the dolphins had to come close to the touch screen and to position themselves using monocular vision (left or right eye) and/or binocular naso-ventral vision. The results showed an ability to associate simple visual forms and auditory information using an underwater touch-screen. Moreover, the subjects showed a spontaneous tendency to use monocular vision. Contrary to previous findings, our results did not clearly demonstrate right eye preference in spontaneous choice. However, the individuals' scores of correct answers were correlated with right eye vision, demonstrating the advantage of this visual field in visual information processing and suggesting a left hemispheric dominance. We also demonstrated that the nature of the presented visual stimulus does not seem to have any influence on the animals' monocular vision choice. PMID- 16246505 TI - Rapid habituation of scan behavior in captive marmosets following brief predator encounters. AB - Scan behavior in 10 captive predator-naive adult black tufted-ear marmosets (Callithrix penicillata) was investigated prior, during and following brief predator encounters (taxidermized oncilla cat -- Leopardus tigrinus) versus neutral stimulus exposures (stuffed toy). For each stimulus, three 9 min home cage trials were conducted > or = 72 h apart. Each trial was divided into three consecutive 3 min intervals: pre-exposure baseline observation, stimulus exposure, and post-exposure observation period. Post-exposure scan duration increased during the first two predator confrontations, while scan frequency increased significantly only after the first. Scan behavior remained constant within the last predator encounter, as it also did within and between the three neutral stimulus exposures. Although marmosets scanned more often and significantly longer after encountering the predator than the neutral stimulus, this response rapidly habituated by the second trial. Therefore, black tufted-ear marmosets in a familiar environment rapidly habituate to brief repeated predator encounters, possibly minimizing anti-predation costs once the degree of a potential threat has been adequately assessed. PMID- 16246504 TI - Dopamine D1 activation shortens the duration of phases in stereotyped grooming sequences. AB - Rats frequently emit grooming actions in a highly stereotyped, syntactic chain in which three distinct phases of facially directed forearm movements are sequentially emitted in a rule-governed pattern and followed by body-directed licking. The present study evaluated the effects of the full dopamine D1 agonist, SKF 81297, and the partial dopamine D1 agonist, SKF 38393, on the duration of individual phases of stereotyped grooming chains. We found that systemic administration of SKF 81297 significantly shortened grooming chain duration. An examination of the fine temporal structure of syntactic grooming chain actions showed that duration changes were correlated with decreased numbers of actions in phases I and IV of the chain. Phases II and III were not changed in duration, although there were some structural distortions introduced. The partial D1 agonist, SKF 38393, had no effect on duration or number of component actions in the grooming chain. Based on these results, we hypothesize that the timing of syntactic grooming phase transitions may involve a D1-mediated internal clock process that is altered by full D1 agonist activation. By this model, SKF 81297 increases the speed of the clock used for the temporal control of grooming actions, and thus shortens phase durations. PMID- 16246506 TI - Sortase A contributes to pneumococcal nasopharyngeal colonization in the chinchilla model. AB - Sortase A (SrtA) is required to anchor neuraminidase, beta-galactosidase, and possibly other LPXTG motif proteins to the pneumococcal cell surface. We examined the role of SrtA in Streptococcus pneumoniae nasopharyngeal (NP) colonization in the chinchilla model. The srtA mutant colonized the nasopharynx at a significantly lower level than the D39 parent strain during the second and third week of the carriage, and was eliminated from nasopharynx one week earlier than the D39 pneumococci. Our data indicate that SrtA contributes to pneumococcal NP colonization in this animal model. PMID- 16246507 TI - Validation protocol and analytical quality in biological monitoring of occupational exposure to antineoplastic drugs. AB - Occupational exposure to antineoplastic cytostatic drugs has been recognized as a potential health hazard since the seventies. Safety guidelines and recommendations have been published in several countries in order to improve operating procedures and keep exposure levels as low as possible. Nevertheless, contamination still occurs. With a view to preventing exposure, standardized authorized methods and a mandatory monitoring system appear to be an essential prerequisite. First of all, reliable analytical methods are necessary, and a wide number of drugs should be monitored, on the basis of the continual change in the treatment protocols. In fact, most of the methods available in the literature were not properly validated, and a lack of details concerning quality assurance is clearly observed. When assessing analytical methods applied for routine testing, validation studies are of utmost importance. Not only performance parameters, such as sensitivity, specificity, LOD and LLQ, should therefore be determined, but also the uncertainty of measurement, which gives a measure of the confidence that can be placed on the results. PMID- 16246508 TI - Biological monitoring of exposure to polycyclic aromatic hydrocarbons by determination of unmetabolized compounds in urine. AB - In this paper we evaluated the possibility to assess occupational exposure to polycyclic aromatic hydrocarbons (PAHs) measuring unmetabolized PAHs in urine. With this aim, 24 road paving (RP) workers, exposed to bitumen fumes, and 6 road construction workers (CW), exposed to diesel exhausts, were investigated. Median personal exposure to low boiling PAHs (from naphthalene to pyrene) during the work shift ranged from 0.5 to 369 ng/m(3), with naphthalene as the most abundant compound. Three urine samples were collected for each worker: baseline (after 2 days of vacation), before- and end-shift samples (in the second part of the work week). The following urinary compounds were measured by headspace-solid phase microextraction GC/MS: naphthalene (U-NAP), acenaphthylene (U-ACY), acenaphthene (U-ACE), fluorene (U-FLE), phenanthrene (U-PHE), anthracene (U-ANT), fluoranthene (U-FLU), pyrene (U-PYR). Urinary PAHs were detected in almost all samples. Median levels for U-NAP, U-PHE, U-PYR and U-FLE in end-shift samples were 82, 48, 54 and 21 ng/L in RP and 69, 14, 24 and 15 ng/L in CW, respectively. Significant differences in the levels of U-PHE, U-FLU and U-PYR were found between RP and CW (p<0.05). Moreover in RP samples the urinary excretion of most analytes increased during the work shift (p<0.05). These results suggest that urinary PAHs may be useful biomarkers of occupational exposure. PMID- 16246509 TI - Mesenteritis precedes vasculitis in the rat mesentery after subacute administration of a phosphodiesterase type 4 inhibitor. AB - Inhibitors of phosphodiesterase type 4 (PDE4) are currently exploited as potent drugs for pulmonary diseases. Some PDE4 inhibitors induce necrotizing panarteritis in the mesentery of rats, comparable to spontaneous polyarteritis nodosa in rats and vascular alterations that are induced by various vasoactive compounds, such as fenoldopam and inhibitors of PDE3. The mechanism of toxicity is unknown. In order to investigate the development of arteritis in the splanchnic vasculature of rats, a time-course study was performed with high doses of a compound (BYK169171), specifically inhibiting PDE4. Rats were treated orally for 1-28 days, and alterations in the mesentery were evaluated by histology, morphometry, and immunohistology. As early as 3 days after the onset of treatment, a mesenteritis was found, characterized by macrophage infiltration, fibroblast proliferation, neovascularization, and loss of adipocytes. Incidence and severity of the mesenteritis were low during the first 2 weeks of treatment, but increased with duration of treatment, finally affecting 2/3 of all animals. A segmental necrotizing panarteritis was detected in some rats treated for 21 or 28 days, but always followed a mesenteritis, whereas many animals with mesenteric inflammation did not have vascular lesions. We postulate that PDE4 inhibitors do not cause a primary vasculitis/arteritis in rats, but induce a non-purulent inflammation as the predominant initial toxic effect in the mesentery. This renders their toxic effect distinct from that of PDE3 inhibitors. PMID- 16246510 TI - Physiologically based pharmacokinetic modelling of human exposure to 2 butoxyethanol. AB - A physiologically based pharmacokinetic (PBPK) model describing the disposition of 2-butoxyethanol (2-BE) was developed in order to predict the urinary concentration of its major metabolite, butoxyacetic acid (BAA) under a range of exposure scenarios. Based on Corley et al. [Corley, R.A., Bormett, G.A., Ghanayem, B.I., 1994. Physiologically based pharmacokinetics of 2-butoxyethanol and its major metabolite, 2-butoxyacetic acid, in rats and humans. Toxicol. Appl. Pharmacol. 129, 61-79], the model included such features as multiple entry routes into the body, varying workload conditions, metabolism in the liver and elimination of free BAA in urine by glomerular filtration and acid transport. A bladder compartment simulating the fluctuations in metabolite concentration in urine caused by micturition formed a novel aspect of the model. Good agreement between model predictions and existing experimental data of total BAA levels in the blood and urine over various exposure conditions were observed. The mechanistically based PBPK model allowed comparison of disparate studies and also enabled the prediction of urinary concentrations of BAA post-shift. By calculating the total amount of BAA, any inter-individual variability in conjugation is taken into account. This led us to conclude that a biological monitoring guidance value should be proposed for total rather than free BAA with a value of 250 mmol/mol of creatinine (post-shift), based on an 8h exposure to 25 ppm 2-BE at resting working conditions. PMID- 16246511 TI - Strategies for improving the functionality of an affinity bioreactor. AB - Heparin employed in extracorporeal blood circulation (ECBC) procedures (e.g. open heart operations) often leads to a high incidence of bleeding complications. Protamine employed in heparin neutralization, on the other hand, can cause severe adverse reactions. We previously developed an approach that could prevent both heparin- and protamine-induced toxic side effects concomitantly. This approach consisted of placing a hollow fiber-based bioreactor device containing immobilized protamine (termed a "protamine bioreactor") at the distal end of the ECBC procedure. This protamine bioreactor would remove heparin after heparin served its anticoagulant purpose in the ECBC device, thereby eliminating heparin induced bleeding risks. In addition, this protamine bioreactor would prevent protamine from entering the patients, thereby aborting any protamine-induced toxic effects. Both in vitro and in vivo studies have successfully demonstrated the feasibility of this approach. Despite promises, early findings also revealed two shortcomings that must be overcome for the protamine bioreactor to be applied clinically. The first drawback was that the cyanate ester linkages, involved in conjugating protamine to the bioreactor device, were unstable and prone to hydrolysis, resulting in the leakage of a significant amount of protamine into circulation during application of the protamine bioreactor. The second deficiency was that the capacity of the protamine bioreactor in heparin removal was rather low, owing to the limited surface area of the hollow fibers for protamine immobilization and subsequently heparin adsorption. In this paper, we present novel strategies to overcome these two limitations. A new conjugation method based on the use of 4-(oxyacetyl)phenoxyacetic acid (OAPA) as the activating reagent was employed to yield stable linkages, via the abundant arginine residues of protamine, onto the hollow fibers. Results showed that while the amount of protamine immobilized on each gram of fibers was relatively comparable between the OAPA and the previous CNBr activation methods (7.45 mg/g versus 7.69 mg/g fibers), there was virtually no detectable leaching of immobilized protamine from the bioreactor by the OAPA method, comparing to 35% leaching of protamine by the previous CNBr method following 72 h of storage of the bioreactor in PBS buffer at 37 degrees C. To improve the capacity and functionality of the protamine bioreactor, two novel approaches were adopted. Long chain and high molecular weight poly-lysine was linked to the hollow fibers, prior to protamine coupling, to create multiple layers of immobilized protamine for subsequent heparin adsorption. In addition, a poly(ethylene glycol) (PEG) chain was inserted between protamine and the hollow fibers to yield a three-dimensional, free dynamic motion for immobilized protamine. Preliminary observations indicated that a four- to five-fold enhancement in heparin adsorption was attained by utilizing each of these new approaches. Aside from their current use, these new strategies can also be employed generically to improve the functionality of any affinity-type bioreactor. Indeed, efforts have been made recently in utilizing these approaches to develop a clinically usable GPIIb/IIIa bioreactor for the treatment of immune thrombocytopenic purpura (ITP)-an autoimmune disease. PMID- 16246513 TI - Skin permeation: the years of enlightenment. AB - Considerable advances in our understanding of the mechanisms by which drugs permeate the skin barrier have been made over the past 60 years. The key publications, which have influenced the field of skin permeation research are highlighted in the present review. The methodologies commonly employed for estimation of skin permeability are discussed as are the mechanisms proposed for skin permeation. The principal findings from the commonly employed in vitro and in vivo models are considered as well as the applications of enhancers and surfactants for optimization of skin delivery. As these studies presaged the emergence of transdermal drug delivery research in the 1970s, early approaches to model and predict dermal and transdermal absorption are also outlined. The published work on skin permeability in this period embodies the fundamental literature sources for consultation by scientists new to and currently engaged in transdermal drug delivery. PMID- 16246512 TI - Optimisation and characterisation of bioadhesive controlled release tetracycline microspheres. AB - A Box-Behnken experimental design was employed to statistically optimise the formulation parameters of a tetracycline microsphere preparation for maximum bioadhesivity and controlled drug release. The quantitative effect of the formulation parameters at different levels on bioadhesion and drug release could be predicted using polynomial equations. A formulation comprising of 3% (w/w) chitosan, 10% (w/w) tetracycline HCl and 9% (w/v) tripolyphosphate was identified for maximising bioadhesivity and obtaining controlled drug release. The optimal microsphere preparation was subsequently characterised in terms of hydration dynamics, release kinetics, antimicrobial activity, thermal properties, morphology and surface pH. Kinetic models revealed that drug release followed Fickian diffusion while textural analysis showed minimal hydration over the test period. Antimicrobial studies showed that the drug concentrations in the in vitro release samples were above the minimum concentration of drug required for inhibition of Staphylococcus aureus growth. Thermal analyses showed a possible interaction between the drug and polymer. Scanning electron microscopy confirmed the integrity of the microspheres and identified the morphological changes following drug release. Surface pH of the microspheres was similar to salivary pH and did not show extremes in changes over the test period. PMID- 16246514 TI - Proteasome dysfunction in mammalian aging: steps and factors involved. AB - Mammalian aging is a natural biological process, determined by both genetic and environmental/stochastic factors, that results in the gradual decline of physiological function and the eventual failure of organism homeostasis. The proteasome is one of the major proteolytic systems of mammalian cells. It is responsible for the degradation of normal proteins as well as of abnormal proteins (like misfolded and oxidized proteins) that tend to accumulate during aging. Impaired proteasome function has been tightly correlated with aging both in vivo and in vitro. Given the fundamental function of proteasome for retaining cellular homeostasis, this review article examines the steps and the factors involved in proteasome dysfunction during mammalian aging. We discuss the proteasome structural organization, its activities and biosynthesis during aging and senescence as well as the genetic and environmental causes of its age dependent alterations. Finally, we provide insights on the possibilities of proteasome activation that may retard the appearance of the senescent phenotype. PMID- 16246515 TI - HIV treatment-related knowledge and self-efficacy among caregivers of HIV infected children. AB - OBJECTIVE: Improvements in treatment-related knowledge and self-efficacy may improve clinical outcomes in HIV-infected populations. We examined whether caregivers' knowledge and self-efficacy was associated with better clinical outcomes and anti-retroviral therapy (ART) adherence among HIV-infected children. METHODS: Caregivers of 77 perinatally HIV-infected children were administered a semi-structured interview which included scales of HIV treatment-related knowledge, adherence self-efficacy and caregiver reports of child medication adherence. RESULTS: While caregivers correctly answered 74% of the knowledge questions, specific misconceptions were noted. Caregivers rated themselves as having high adherence self-efficacy, but were least confident in their ability to adhere to ART if it caused side effects or might result in social disclosure. Higher caregiver treatment-related knowledge and self-efficacy were associated with better clinical outcomes, as measured by viral load and CD4+ count, but not with caregiver reports of ART adherence. CONCLUSION/DISCUSSION: While additional factors may be important determinants of adherence to ART in this population, improving the HIV treatment-related knowledge and self-efficacy of caregivers may help to improve the clinical outcomes of HIV-infected children. PRACTICE IMPLICATIONS: Care providers should address both misconceptions that might serve to undermine adherence as well as factors that limit patients' self-efficacy in adhering to treatment, prior to the initiation of ART and throughout the course of treatment. PMID- 16246516 TI - Results of sentinel node biopsy not affected by previous excisional biopsy. AB - AIMS: Sentinel lymph node (SN) biopsy has been validated in the treatment of breast carcinoma. Patients with previous excisional biopsy are regarded as ineligible for SN biopsy. We evaluated the results of SN biopsy for this group of patients based on confirmatory axillary lymph node dissection. PATIENTS AND METHODS: From April 1997 all 88 patients with stage T(1-3) breast cancer who had previously undergone diagnostic excisional biopsy followed by complete axillary lymph node dissection, were enrolled into a prospective study to determine the validity of the sentinel node procedure. RESULTS: Lymphoscintigraphy visualized one or more axillary hot spots in 84/88 patients. A successful SN biopsy was performed in 87 patients. Complete axillary lymph-node dissection showed no false negative SN biopsy among the 87 SN procedures. CONCLUSION: SN biopsy is a reliable and safe method following excisional biopsy as is confirmed by completion axillary lymph node dissection. Therefore, patients with previous excisional biopsy are eligible for sentinel node procedure and can be spared unnecessary axillary lymph node dissection. PMID- 16246517 TI - Prediction of the risk of hepatic failure in patients with portal vein invasion hepatoma after hepatic resection. AB - AIM: Hepatic failure can develop after curative hepatectomy in patients with a hepatocellular carcinoma (HCC) invading the portal vein, because of cirrhosis and excessive tissue loss. This study aimed to identify the risk factors for hepatic failure in such patients. METHOD: Forty patients with an HCC invading the portal vein underwent curative hepatectomy from January 1995 to June 2003. Eight patients developed hepatic failure and died within 3 months. Possible risk factors for this were analysed using univariate and multivariate regression. These included the liver function index, surgical blood loss, tumour pattern, portal hypertension, estimated residual liver volume measured by computed tomography (ERLV(CT)) and estimated residual liver volume using the indocyanine green (ICG) retention rate at 15 min (ERLV(ICG15)). RESULTS: The ERLV(CT) smaller than the ERLV(ICG15) and presence of portal hypertension were independent risk factors for post-hepatectomy hepatic failure. CONCLUSION: Having portal vein invasion HCC with portal hypertension or an ERLV(CT) less than an ERLV(ICG15) are significant predictors of post-hepatectomy hepatic failure. These factors are important considerations for patients with portal vein invasion HCC who could undergo curative hepatic resection. PMID- 16246518 TI - A case of bilateral breast cancer with Peutz-Jeghers syndrome. PMID- 16246519 TI - The effect of jaundice on the generation of anti-gastrin antibodies in G17DT immunized patients with advanced pancreatic cancer. AB - AIM: The aim of this study was to determine the ability of G17DT to generate anti gastrin antibodies in jaundiced patients with biliary obstruction due to advanced pancreatic cancer. METHODS: G17DT was administered to 41 patients with advanced pancreatic adenocarcinoma by intramuscular (i.m.) injection at a dose of 250mcg at weeks 0, 1 and 3 of the study. RESULTS: Thirty-five of 41 patients participating in the study were categorized as responders in terms of their gastrin-17 antibody response. There was no correlation between the maximum G17 antibody response and the bilirubin level at either week 0 or week 12. The median survival of patients from the time of the first injection of G17DT was 204 days with 25% of patients surviving for or=305 days. CONCLUSION: This study shows that G17DT administered to jaundiced patients with advanced pancreatic cancer is immunogenic and well tolerated. PMID- 16246520 TI - Expression of 3alpha- and 3beta-hydroxy steroid dehydrogenase mRNA in COCs and granulosa cells determines Zearalenone biotransformation. AB - Zearalenone (ZEA) is a mycoestrogen found in diverse food and feed materials, particularly in corn and small grains. Following ingestion, the parent zearalenone is converted predominantly into alpha-zearalenol (alpha-ZOL) and beta zearalenol (beta-ZOL) by hepatic hydroxy steroid dehydrogenases (HSD). The present study demonstrated by standard RT-PCR the expression of 3alpha- and 3beta HSD also in porcine cumulus oocyte complexes (COCs) and granulosa cells isolated form cumulus oocyte complexes. Analysis of the rate of bioconversion of zearalenone (ZEA) by the cultured granulose cells showed the extra-hepatic production of both hydroxy metabolites of ZEA with alpha-ZOL being the dominating metabolites as previously observed in incubations with liver microsomes. The endogenous steroids 5alpha-dihydrotestosterone (5alpha-DHT), and progesterone (PGTN), both known substrates for 3alpha-HSD inhibited the conversion of ZEA into alpha-ZOL. In the presence of pregnelonone (PGN), a major substrate for 3beta-HSD only a slight inhibitory effect on the apparent beta-ZOL formation could be observed. In conclusion, these data indicate that both 3alpha- and 3beta-HSDs are expressed in porcine COCs and GCs, whereas the biotransformation experiments confirm the involvement of these enzymes in the extra-hepatic biotransformation of ZEA. PMID- 16246521 TI - Piggy-BACing the human genome II. A high-resolution, physically anchored, comparative map of the porcine autosomes. AB - Using the INRA-Minnesota porcine radiation hybrid panel, we have constructed a human-pig comparative map composed of 2274 loci, including 206 ESTs and 2068 BAC end sequences, assigned to 34 linkage groups. The average spacing between comparative anchor loci is 1.15 Mb based on human genome sequence coordinates. A total of 51 conserved synteny groups that include 173 conserved segments were identified. This radiation hybrid map has the highest resolution of any porcine map to date and its integration with the porcine linkage map (reported here) will greatly facilitate the positional cloning of genes influencing complex traits of both agricultural and biomedical interest. Additionally, this map will provide a framework for anchoring contigs generated through BAC fingerprinting efforts and assist in the selection of a BAC minimal tiling path and assembly of the first sequence-ready map of the porcine genome. PMID- 16246522 TI - Enhanced expression of stem cell antigen-1 (Ly-6A/E) in lymphocytes from lupus prone mice correlates with disease severity. AB - B6.Sle1 mice, congenic for the NZM2410-derived lupus susceptibility locus, Sle1 on chromosome 1 exhibit many of the features seen in human lupus including activated lymphocytes and high titers of antinuclear autoantibodies. Among the different surface molecules that were aberrantly expressed on the B6.Sle1 lymphocytes was Ly-6A/E. Splenic B- and T-lymphocytes but not myeloid cells from B6.Sle1 mice exhibited enhanced levels of Ly-6A/E compared to B6 controls. In particular, MZ B cells, GC B cells and B-cell blasts expressed the highest levels of Ly-6A/E in both strains, with the levels being even higher on B6.Sle1 derived cells. Following stimulation with LPS or anti-IgM, there was a profound up regulation in Ly-6A/E, particularly on MZ B cells and B-cell blasts. CD4 and CD8 T cells also up-regulated Ly-6A/E after stimulation with anti-CD3 and anti-CD28. These studies were extended to additional autoimmune strains including B6.Sle3, B6.Sle1.lpr and BXSB. Importantly, Ly-6A/E levels on lymphocytes were commensurate with the degree of disease exhibited by these lupus strains. Finally, it appears that increased interferon levels, in addition to antigen receptor stimulation, may also be a factor accounting for elevated Ly-6A/E in lupus. Given these observations it is important to elucidate the functional role of Ly-6A/E in lupus in future studies. PMID- 16246523 TI - Anti-p80 coilin autoantibodies react with a conserved epitope and are associated with anti-DFS70/LEDGF autoantibodies. AB - p80 coilin is a nuclear autoantigen that strongly accumulates in Cajal bodies (CB) and is considered a marker for CBs. No clear clinical features have been associated with anti-p80 autoantibodies, and the epitopes recognized by these antibodies are entirely unknown. We determined the epitope specificity for 34 anti-p80 coilin-positive sera from Japanese patients to better understand its clinical significance. Western blotting and immunoprecipitation analysis revealed that the autoepitope of p80 coilin was restricted to a single region encompassing amino acid residues 421-576 from the C-terminus of p80 coilin. Additionally, sera from 30 of 34 (88%) patients with anti-p80 coilin antibody titers were also positive for anti-DFS70/LEDGF antibody. This autoantibody is sometimes found in patients with some allergic diseases but rarely in patients with connective tissue disease (CTD). Other similar characteristics between these two autoantibodies were also noted. In contrast, few anti-p80 coilin positive patients were positive for CTD-marker antibodies, such as anti-Sm, anti-dsDNA, and anti-SS-A. We conclude that the presence of anti-p80 coilin autoantibodies is not associated with any particular clinical syndromes nor is it diagnostic of any conditions. PMID- 16246524 TI - Insulin alleles and autoimmune regulator (AIRE) gene expression both influence insulin expression in the thymus. AB - It is well established that the polymorphisms at the 5' of the insulin gene (IDDM2) confers susceptibility to type 1 diabetes, probably by modifying the level of insulin expression in the thymus that in turn influences immunological tolerance to insulin as self-antigen. AIRE is a transcription regulator which controls the expression of many peripheral antigens within the thymus, among them insulin. Results presented here confirm that insulin gene copies from both parental chromosomes are expressed in human thymus and that IDDM2 class III protective alleles are indeed associated with a higher level of insulin message expression. However, differences in insulin mRNA expression among different thymi were far wider than those determined by the class I and class III insulin gene alleles and maintained a clear correlation with AIRE expression. These results confirm the effect of IDDM2 alleles on insulin expression in the thymus, but suggest that the levels of AIRE may exert a stronger influence than IDDM2 alleles themselves. PMID- 16246525 TI - DR.lyp/lyp bone marrow maintains lymphopenia and promotes diabetes in lyp/lyp but not in +/+ recipient DR.lyp BB rats. AB - Lymphopenia is due to a frameshift mutation in Gimap5 on rat chromosome 4 and is linked to type 1 diabetes in the diabetes prone (DP) BB rat. The hypothesis that bone marrow derived cells confer the lymphopenia phenotype was tested by reciprocal bone marrow transplantation in 40-day-old lethally irradiated diabetes resistant (DR) congenic DR.lyp/lyp (lymphopenia and diabetes) and DR.+/+ (no lymphopenia and no diabetes) rats. In two independent series of transplants, all DR.lyp/lyp rats (n=5 and 4) receiving DR.lyp/lyp bone marrow retained lymphopenia and developed insulitis (5/5 and 4/4) as well as diabetes in some (2/5 and 3/4). Both DR.+/+ and DR.lyp/lyp rats receiving DR.+/+ bone marrow cells as well as DR.+/+ rats receiving DR.lyp/lyp bone marrow cells showed no lymphopenia or diabetes. In accordance with earlier studies in non-congenic BB rats, the DR.+/+ rats receiving DR.lyp/lyp bone marrow cells recapitulated an intermediary phenotype rather than the +/+ or lyp/lyp phenotypes. Our data demonstrate that BBDP rat lymphopenia and diabetes are transferred by bone marrow transplantation to syngeneic DR.lyp/lyp but not DR.+/+ recipients. The intermediary recapitulation of DR.lyp/lyp T cells in recipient DR.+/-/+/- rats suggests that radiation resistant +/-/+/- T cells, the Gimap5 mutation in bone marrow cells, or both may not support the development of lymphopenia. PMID- 16246527 TI - Pemphigus vulgaris-IgG reduces the desmoglein 3/desmocollin 3 ratio on the cell surface in cultured keratinocytes as revealed by double-staining immunoelectron microscopy. PMID- 16246526 TI - Memory tests in first-degree adult relatives of schizophrenic patients: a meta analysis. AB - BACKGROUND: Memory deficits have been clearly demonstrated in schizophrenic patients. However, studies of memory performances in their relatives compared to normal controls provide conflicting results. A meta-analysis was carried out to synthesize all the published data. Unlike previous meta-analyses, which were based on composite scores, we analyzed each memory test separately. This prevents theoretically questionable choices in grouping variables, leads to results with clearer implications for applied research (e.g. the best choice of a test according to its sensitivity) and is more productive in suggesting explanatory hypotheses. METHOD: We initially selected 77 potentially relevant articles, but only 19 met our inclusion criteria. These articles provided data on eight different tasks, from five different memory tests: four tests from the Wechsler Memory Scale (WMS) and the California Verbal Learning Test (CVLT). For each task, we assessed data homogeneity, identified the outliers if any and then estimated effect sizes and tested publication bias using funnel plots. RESULTS: Adult relatives of schizophrenic patients were significantly impaired on most, but not all, tasks. The largest deficits were observed for the verbal paired associates test, the logical stories the digit span forward test and the digit span backward test. We found no significant differences in tasks of delayed recall, when deficits in immediate conditions (reflecting encoding) were taken into account. CONCLUSIONS: Adult relatives of schizophrenic patients have wide but not severe memory impairments. The size of estimated effects suggests that encoding processes are impaired, whereas storage and retrieval processes are relatively unaffected. PMID- 16246529 TI - Allopregnanolone produces hyperphagia by reducing neophobia without altering food palatability. AB - The neurosteroid allopregnanolone may increase feeding by altering food palatability; however, it may also increase feeding by reducing anxiety (neophobia). Moreover, it is unclear whether this induced hyperphagia is selective to safe, palatable foods only. Male rats were injected with allopregnanolone 20 min prior to behavioral testing. The taste reactivity test was used to examine possible shifts in the palatability of a 0.3 M sucrose solution. A lickometer was used to monitor intake and licking of either a sucrose or sucrose-quinine solution. Sucrose palatability was not enhanced; however, allopregnanolone significantly increased sucrose intake and licking on Test Day 1 when the solution was novel, but not on Test Day 2 when the solution was familiar. Sucrose-quinine intake was not enhanced. Allopregnanolone-induced hyperphagia is not a result of altered sucrose palatability, but rather reflects a reduction in the neophobia elicited by a novel solution; an effect that further seems to be selective to safe, palatable foods. PMID- 16246530 TI - The effect of tricyclic antidepressants, selective serotonin reuptake inhibitors (SSRIs) and newer antidepressant drugs on the activity and level of rat CYP3A. AB - The aim of the present study was to investigate the influence of tricyclic antidepressants (TADs: imipramine, amitriptyline, clomipramine, and desipramine), selective serotonin reuptake inhibitors (SSRIs: fluoxetine and sertraline) and novel antidepressant drugs (mirtazapine and nefazodone) on the activity of CYP3A measured as a rate of testosterone 2beta- and 6beta-hydroxylation. The reaction was studied in control liver microsomes in the presence of the antidepressants, as well as in microsomes of rats treated intraperitoneally (i.p.) for 1 day or 2 weeks with pharmacological doses of the drugs (imipramine, amitriptyline, clomipramine, nefazodone 10 mg kg(-1) i.p.; desipramine, fluoxetine, sertraline 5 mg kg(-1) i.p.; mirtazapine 3 mg kg(-1) i.p.), in the absence of the antidepressants in vitro. The investigated antidepressants added to control liver microsomes produced some inhibitory effects on CYP3A activity, which were very weak (most of TADs, K(i)=145-212 microM), modest (clomipramine and sertraline, K(i)=67.5 and 62 microM, respectively) or moderate (nefazodone and fluoxetine, K(i)=42 and 43 microM, respectively). Mirtazapine did not display this kind of properties. One-day exposure of rats to TADs substantially decreased the activity of CYP3A in liver microsomes, which was maintained during chronic treatment. The observed decreases in the enzyme activity were in contrast to the increased CYP3A protein level found after chronic treatment with TADs. On the other hand, sertraline increased the activity of the enzyme after its prolonged administration and its effect correlated positively with the observed elevation in CYP3A protein level. Fluoxetine, mirtazapine and nefazodone did not change the activity of CYP3A in liver microsomes after their administration to rats. Three different mechanisms of the antidepressants-CYP3A interaction are postulated: 1) a direct inhibition of CYP3A by nefazodone, SSRIs and clomipramine, shown in vitro, with the inhibitory effect of nefazodone being the strongest, but weaker than the effects of this drug on human CYP3A4; 2) in vivo inhibition of CYP3A produced by 1 day and maintained during chronic treatment with TADs, which suggests inactivation of the enzyme by reactive metabolites; 3) in vivo induction by sertraline of CYP3A produced only by chronic treatment with the antidepressant, which suggests its influence on the enzyme regulation. PMID- 16246531 TI - Multiple-sample probe for solid-state NMR studies of pharmaceuticals. AB - Solid-state NMR spectroscopy (SSNMR) is an extremely powerful technique for the analysis of pharmaceutical dosage forms. A major limitation of SSNMR is the number of samples that can be analyzed in a given period of time. A solid-state magic-angle spinning (MAS) probe that can simultaneously acquire up to seven SSNMR spectra is being developed to increase throughput/signal-to-noise ratios. A prototype probe incorporating two MAS modules has been developed and spectra of ibuprofen and aspirin have been acquired simultaneously. This version is limited to being a two-module probe due to large amounts of space required for the tuning elements located next to the MAS modules. A new probe design incorporating coaxial transmission lines and smaller MAS modules has been constructed. This probe allows for close proximity of the MAS modules (within 3 cm), adequate proton decoupling power (>50 kHz), and the capability of remote tuning and sample changing. Spectra of hexamethylbenzene (HMB) have been acquired and show signal to-noise ratios comparable to existing SSNMR probes. Adamantane line widths are also comparable to conventional probe technology. Decoupling powers of 70 kHz have been achieved using a MAS module suitable for 3 cm spacing between modules. Remote tuning has also been achieved with this new coaxial transmission line design. This probe design can be easily scaled to incorporate multiple MAS modules, which is a limitation of the previous design. The number of modules that can be incorporated is only limited by the number of transmission lines that will fit in a cross-sectional diameter of the bore and the axial field length of the magnet. PMID- 16246532 TI - Attentional shifts towards an expected visual target alter the level of alpha band oscillatory activity in the human calcarine cortex. AB - Neuronal operations associated with the top-down control process of shifting attention from one locus to another involve a network of cortical regions, and their influence is deemed fundamental to visual perception. However, the extent and nature of these operations within primary visual areas are unknown. In this paper, we used magnetoencephalography (MEG) in combination with magnetic resonance imaging (MRI) to determine whether, prior to the onset of a visual stimulus, neuronal activity within early visual cortex is affected by covert attentional shifts. Time/frequency analyses were used to identify the nature of this activity. Our results show that shifting attention towards an expected visual target results in a late-onset (600 ms postcue onset) depression of alpha activity which persists until the appearance of the target. Independent component analysis (ICA) and dipolar source modeling confirmed that the neuronal changes we observed originated from within the calcarine cortex. Our results further show that the amplitude changes in alpha activity were induced not evoked (i.e., not phase-locked to the cued attentional task). We argue that the decrease in alpha prior to the onset of the target may serve to prime the early visual cortex for incoming sensory information. We conclude that attentional shifts affect activity within the human calcarine cortex by altering the amplitude of spontaneous alpha rhythms and that subsequent modulation of visual input with attentional engagement follows as a consequence of these localized changes in oscillatory activity. PMID- 16246533 TI - Construction and deconstruction of PLL/DNA multilayered films for DNA delivery: effect of ionic strength. AB - Through the layer-by-layer (LbL) self-assembly technique, DNA was incorporated into the multilayered films with poly-l-lysine (PLL). The effect of ionic strength on the construction and deconstruction of the PLL/DNA films was investigated. It was found that the salt concentration of the deposition solution had a significant effect on the construction of the films, which might attribute to the effect of salt ions on the conformation of polyelectrolytes and interaction between PLL and DNA molecules. A salt-induced deconstruction of the PLL/DNA films was observed. The extent of the deconstruction increased with the salt concentration in the incubation solution. The mechanism of the deconstruction was discussed. Taking the advantages of the LbL technique, the erasable PLL/DNA films could deposit onto a variety of surfaces, such as vascular stent, intervention catheter and tissue engineering scaffold, to serve as a novel DNA delivery system. PMID- 16246534 TI - The effect of solvent polarity on the accumulation of leachables from pharmaceutical product containers. AB - Material/water equilibrium interaction constants (E(b)) were determined for 12 organic model solutes and a plastic material used in pharmaceutical product containers (non-PVC polyolefin). An excellent correlation was obtained between the measured interaction constants and the organic solute's octanol/water partition coefficient. The effect of solvent polarity on E(b) was assessed by examining the interaction between the plastic and selected model solutes in binary ethanol/water mixtures. In general, logE(b) could be linearily related to the polarity of the ethanol/water mixture. This information, coupled with the interaction model, was used to estimate the levels to which container leachables could accumulate in contacted solutions. Such estimates were made for six known leachables of the polyolefin material and compared to the leachable's measured accumulation levels in binary ethanol/water systems. In general, the accumulation level of the leachables increased with increasing solution polarity. For most of the leachables, the measured accumulation level was less than the calculated levels, suggesting that equilibrium was not achieved in the leaching portion of this study. This lack of equilibrium is attributable to the layered structure of the material studied, as such layering retards the migration of the leachables that are derived from the material's non-solution contact layers. PMID- 16246535 TI - The effect of crystal morphology and mill type on milling induced crystal disorder. AB - Milling is a key process in the preparation of many solid dosage forms. One possible milling induced change is the production of small levels of disorder or amorphous material found predominantly at the surface of a powder, which could lead to significant chemical and physical instability. The influence of crystal habit on this change was investigated using beta-succinic acid, in plate like and needle like morphologies. beta-Succinic acid crystals with these habits were processed in a ball mill and a jet mill. SEM images indicated jet milled material was finer than the ball milled product. Powder X-ray diffraction of the milled powders revealed an amorphous halo at lower angles and peak broadening suggesting disorder though this could not be quantified accurately. In addition, a partial conversion during milling to the alpha form was noted. Quantitation of the alpha form in the milled powders indicated it was present at <2% (w/w). Plate and needle shaped particles had similar heats of solution pre-milling, however, all milled powders had lower heats of solution compared to the unmilled powers. The contribution of the alpha polymorph to the lower heats of solution was calculated to be insignificant. Therefore, the reduced heat of solution is attributed to a loss in crystallinity. The largest decreases were seen in the plate like morphology. These findings suggest that beta-succinic acid crystals with plate like morphology are more prone to crystallinity loss on milling compared to the needle like morphology. The mill type has also been shown to influence the final crystallinity. PMID- 16246537 TI - Evaluation of the potential of air jet milling of solid protein-poly(acrylate) complexes for microparticle preparation. AB - It was the aim of this study to evaluate the potential of air jet milling for the preparation of protein-loaded microparticles in industrial quantities. The model protein horseradish peroxidase was incorporated via co-precipitation in carbomer (NaC934P) (1:100) and a poly(methacrylate) (Eudragit L100-55) (1:100) used as carrier matrix. Co-precipitation of the model protein and each polymer in aqueous solution was achieved either by a pH-shift or by the addition of various non solvents. Dried protein/polymer complexes (desiccator under vacuumization at 4 degrees C with silica blue gel) were ground with an air jet mill and resulting microparticles were investigated regarding protein load, remaining protein activity, size distribution and shape. Results of this study showed that the polymer used and the method of co-precipitation has a great impact on protein load. Using carbomer a maximum protein load of 60 +/- 1% was achieved, whereas in case of Eudragit L100-55 the maximum was 78 +/- 5% (means +/- SD; n = 3-4). Using petroleum ether, isopropanol or tetrahydrofurane as non-solvents led to significantly higher protein loads than a pH-shift from 7 to 5, 4 and 3.5, respectively. Determination of the remaining protein activity after milling showed, that the grinding air pressure (GAP) has a major impact on protein stability. In case of Eudragit L100-55 at a GAP of 4.5 bar peroxidase activity was almost completely lost, whereas 42 +/- 1% loss in activity was determined at a GAP of 2.5 bar. The mean particle size of protein/carbomer and protein/poly(methacrylate) particles was determined to be 3.6-5.2 and 4.5-8.7 microm at a GAP of 2.5 bar and 2.7-3.1 and 2.4-3.1 microm at a GAP of 4.5 bar, respectively. Generally, 90% of all particles were in the range of 3-16 microm. All particles were of spherical shape exhibiting a non-porous surface. According to these results, air jet milling seems to represent a novel method for the large scale production of protein drug loaded microparticles. PMID- 16246536 TI - Drug permeability across a phospholipid vesicle based barrier: a novel approach for studying passive diffusion. AB - The aim of this study was to develop a novel predictive medium-throughput screening method for drug permeability, with use of a tight barrier of liposomes on a filter support. To our knowledge no one has succeeded in depositing membrane barriers without the use of an inert solvent such as hexadecane. The first part of the study involved development of a protocol for preparation of these barriers, which were made of liposomes from egg phosphatidylcholin in phosphate buffer pH 7.4 with 10 % (v/v) ethanol. The liposomes were deposited into the pores and onto the surface of a filter support (mixed cellulose ester) by use of centrifugation. Solvent evaporation and freeze-thaw cycling were then used to promote fusion of liposomes. A tight barrier could thus be obtained as shown with calcein permeability and electrical resistance. In the second part of the study the model was validated using 21 drug compounds, which cover a wide range of physicochemical properties and absorption (F(a)) in humans (13-100%). The drug permeation studies were carried out at room temperature with phosphate buffer (pH 7.4) in both acceptor and donor chambers. The apparent permeability coefficients obtained from the phospholipid vesicle based model correlated well with literature data on human absorption in vivo, which suggests that its performance is adequate and that the method is suitable for rapid screening of passive transport of new chemical entities. The results obtained from our model were compared with polar surface area (PSA) and experimental logD and with results obtained by established permeability screening methods such as immobilized liposome chromatography (ILC), the PAMPA models and the Caco-2 model. Our approach seems to model the in vivo absorption better than PSA, experimental logD, the ILC and PAMPA models, when similar conditions are used as in our assay, and equally well as the Caco-2 model and the Double Sink PAMPA (DS-PAMPA) model. PMID- 16246538 TI - Development of an in vitro/in vivo correlation for lipid formulations of EMD 50733, a poorly soluble, lipophilic drug substance. AB - PURPOSE: To develop lipid semisolid formulations of EMD 50733, a poorly soluble, neutral drug candidate and to develop an in vitro-in vivo correlation for these formulations using the dog as the in vivo model. METHODS: The model drug, EMD 50733, (with BCS Class II properties) was dissolved in molten lipid/surfactant mixtures and the melt was filled into hard capsules and allowed to re-solidify at room temperature. The dissolution profiles in bio-relevant dissolution media and the bioavailability in dogs were measured and compared to that of a standard formulation consisting of a lactose/drug mixture. RESULTS: The best results with respect to dissolution, stability upon storage and bioavailability were obtained with a formulation that contained a commercially available lipid mixture (Gelucire 44/14) and a solubilizing agent (2-vinylpyrrolidone). With this formulation it was possible to dissolve a typical drug dose in a fill volume suitable for a #0 capsule. Additionally, surface tension measurements showed that the formulation formed micelles during dissolution in aqueous media: the molecular dispersion of the drug in this self-micelle forming system is postulated to protect the drug from precipitation in vivo as well as in vitro. For other formulations tested, neither the in vitro nor the in vivo performance indicated sufficient drug solubilizing properties. CONCLUSION: To achieve adequate and reliable dissolution of poorly soluble drugs in vivo, lipid excipients should not only have appropriate solubilizing properties for the drug in the formulation, but should also assist in maintaining drug in solution during release in the GI tract. PMID- 16246539 TI - Systemic CD4+ T-cell activation is correlated with FEV1 in smokers. AB - The inflammation of the lungs in chronic obstructive pulmonary disease (COPD) is characterised by increased numbers of macrophages, neutrophils and T-cells. Decline in lung function in these patients has been correlated to the number of CD8+ T-cells present in the lung as well as to a decline in the ratio of CD4+/CD8+ T-cells. Although systemic components are likely to be present, circulating lymphocyte populations in COPD patients have not been well characterised. This study aimed at correlating lung function to expression of five different T-cell activation markers on peripheral blood CD4+ and CD8+ T cells in COPD patients and matched smokers. Furthermore, proportions of lymphocyte populations and degree of systemic T-cell activation in COPD patients were compared to that in smokers and never-smokers. Peripheral blood lymphocytes from six never-smokers, eight smokers and 17 smokers with COPD were analysed using flowcytometry. The number of lymphocytes per millilitre was higher in smokers than in never-smokers. No differences were found between the three groups in regard to proportions of lymphocyte populations, but the number of CD4+ T cells in smokers was higher than in both never-smokers and COPD patients. The degree of T-cell activation was similar in all patient groups; however, a clear correlation between CD69 expression on CD4+ T-cells and lung function (FEV(1)% of predicted) was found when examining current smokers, with or without COPD. Elevated numbers of CD69+ CD4+ T-cells in blood thus seem to be protective against airway obstruction in smokers while still exposed to cigarette smoke, the main inducer of COPD. PMID- 16246540 TI - Obstructive sleep-disordered breathing and plasma levels of vascular endothelial growth factor in children. AB - Vascular endothelial growth factor (VEGF) may be one of the pathophysiologic links in the association between obstructive sleep apnea-hypopnea and cardiovascular disease. Morning serum VEGF levels are increased in children with obstructive sleep apnea. However, release of VEGF by platelets and leukocytes during blood clotting may affect its concentration in serum. In the present study, VEGF levels were measured in children with and without habitual snoring using plasma specimens. Evening and morning plasma VEGF concentrations were determined in: (i) 20 children with habitual snoring and apnea-hypopnea index (AHI)5 episodes/h (median age 5; range 1.9-13 years); (ii) 55 children with snoring and AHI<5 episodes/h (median age 6; 2-13 years); and (iii) 25 controls without snoring (median age 6.5; 3-13 years). No differences were identified between the three study groups regarding evening [median 2.5 (range 2.5-174.5) versus 22.5 (2.5-159.4) versus 26.8 (2.5-108) pg/mL; P>0.05] and morning VEGF levels [median 7.7 (range 2.5-120.5) versus 25.1 (2.5-198.4) versus 48.4 (2.5 147.7) pg/mL; P>0.05]. AHI and % sleep time with oxygen saturation of hemoglobin less than 90% were not significant predictors of log-transformed morning VEGF concentrations (P>0.05). In summary, both evening and morning plasma VEGF levels were similar in children with obstructive sleep-disordered breathing of variable severity and in controls without snoring. VEGF may not play an important pathophysiologic role in all cases of obstructive sleep-disordered breathing in childhood. PMID- 16246541 TI - An overview on olive mill wastes and their valorisation methods. AB - Olive mill wastes represent an important environmental problem in Mediterranean areas where they are generated in huge quantities in short periods of time. Their high phenol, lipid and organic acid concentrations turn them into phytotoxic materials, but these wastes also contain valuable resources such as a large proportion of organic matter and a wide range of nutrients that could be recycled. In this article, recent research studies for the valorisation of olive mill wastes performed by several authors were reviewed: second oil extraction, combustion, gasification, anaerobic digestion, composting and solid fermentation are some of the methods proposed. Special attention was paid to the new solid waste generated during the extraction of olive oil by the two-phase system. The peculiar physicochemical properties of the new solid waste, called two-phase olive mill waste, caused specific management problems in the olive mills that have led to the adaptation and transformation of the traditional valorisation strategies. The selection of the most suitable or appropriate valorisation strategy will depend on the social, agricultural or industrial environment of the olive mill. Although some methods are strongly consolidated in this sector, other options, more respectful with the environment, should also be considered. PMID- 16246542 TI - SPR-based immunosensor for the CRP detection--a new method to detect a well known protein. AB - The c-reactive protein (CRP) is one of the significant human blood serum markers for inflammatory processes. The serum presence of this hepatic approximately 115 kDa protein of five identical subunits accompanies several diseases (e.g. CVD, inflammatory bowel diseases) and is nowadays detected by high-sensitivity ELISA assays in blood serum. To enable CRP detection in other matrices, an SPR-based (surface plasmon resonance) immunosensor for the CRP detection has been established. A linear detection range of 2-5 microg CRP per ml was found, using two different antiCRP antibodies (monoclonal, IgG) for CRP trapment and detection. Furthermore, the kinetic antibody association and dissociation constants of one antibody (antiCRP, clone C2) could be determined. PMID- 16246543 TI - A low-density oligonucleotide array study for parallel detection of harmful algal species using hybridization of consensus PCR products of LSU rDNA D2 domain. AB - A low-density oligonucleotide array approach based on the hybridization of consensus PCR products of LSU rDNA was developed in order to simultaneously detect various harmful algae. A set of oligonucleotide probes for the hybridization of specific LSU rDNA D2 regions was developed for the identification of 10 representative harmful microalgae. Each probe was spotted onto a streptoavidin-coated glass slide by pipetting. Universal primers were designed within the conserved regions adjacent to the D2 regions of all harmful algae and used to PCR amplify the complete D2 regions. The PCR products were hybridized to the oligonucleotides arrayed on the slide. The array produced unique hybridization patterns for each species of harmful algae and allowed us to differentiate the closely related species. Furthermore, we were able to simultaneously detect several predominant HAB species from a mixture of culture strains and from a natural sample. These results show that DNA microarray can be a new technical platform for parallel discrimination of harmful algae and has great potential to alter the manner in which researchers monitor these microorganisms. PMID- 16246544 TI - Genomes and evolution: the power of comparative genomics. PMID- 16246545 TI - Randomised phase II study of standard versus chronomodulated CPT-11 plus chronomodulated 5-fluorouracil and folinic acid in advanced colorectal cancer patients. AB - In this study, a randomised phase II trial explored the effects of 6-h chronomodulated CPT-11 infusion in advanced colorectal cancer patients. Sixty eight pre-treated patients were randomly assigned to CPT-11 administered at 180 mg/m2 on day 1, by 1-h infusion (Arm A) or 6-h sinusoidal infusion with peak timing at 5:00 a.m. (Arm B). All patients also received chronomodulated folinic acid/5-fluorouracil (FA/5-FU). Patients in Arm B obtained a 25.7% response rate for 7.0 months duration, a progression-free survival for 8.0 months and a median survival of 28 months. The same data in Arm A were 18.2%, 4.5, 6.0 and 18 months, respectively. No differences in drugs dose-intensity or increased toxicity with prolonged chronomodulated infusion were detected. Major grade 3-4 toxicity was diarrhoea: 10 patients in Arm A and 13 in Arm B. In conclusion, this study has shown that chronomodulated infusion of CPT-11 and FA/5-FU is safe, active and can be integrated with oxaliplatin (EORTC 05011) for the treatment of advanced colorectal cancer. PMID- 16246547 TI - Synthesis and antibacterial activity of substituted 1,2,3,4-tetrahydropyrazino [1,2-a] indoles. AB - A series of substituted 1,2,3,4-tetrahydropyrazino [1,2-a] indole derivatives have been synthesized and tested against the Gram positive and Gram negative strains of bacteria namely Staphylococcus aureus (MTCCB 737), Salmonella typhi (MTCCB 733), Pseudomonas aeruginosa (MTCCB 741), Streptomyces thermonitrificans (MTCCB 1824) and Escherichia coli (MTCCB 1652). All synthesized compounds showed mild to moderate activity. However, compounds 4d-f were found to have potent activity against pathogenic bacteria used in the study. Their MIC ranged from 3.75 to 60 microg/disc. In vitro toxicity tests demonstrated that toxicity of 4d f was not significantly different than that of gentamycin. However, at higher concentration (1000-4000 microg/ml) difference was highly significant. PMID- 16246546 TI - Coordinate expression of the PI3-kinase downstream effectors serum and glucocorticoid-induced kinase (SGK-1) and Akt-1 in human breast cancer. AB - The phosphatidylinositol 3-kinase (PI3-K) signalling pathway has been implicated in breast cancer development and resistance to therapy. Akt-1 and serum and glucocorticoid-induced kinase-1 (SGK-1) are homologous kinases which are important downstream effectors of PI3-K signalling. We sought to determine the individual expression patterns of these two kinases in order to better understand their respective roles in PI3-K signalling in breast cancer. To this end, we examined the expression of both p-Akt-1 and SGK-1 in 40 breast cancers. p-Akt-1 expression was seen in 58% of tumour samples, while SGK-1 overexpression was detected in 48%. Interestingly, a highly significant association was found between the expression of p-Akt-1 and SGK-1 (P=0.002), suggesting complementary physiological functions in PI3-K signalling. This finding is consistent with recent genetic data from Caenorhabditis elegans suggesting that both SGK-1 and Akt-1 are required for signalling downstream of insulin receptor activation. PMID- 16246548 TI - Design, synthesis, and biological evaluation of the N-diarylalkenyl piperidinecarboxylic acid derivatives as GABA uptake inhibitors (I). AB - Twenty novel N-diarylalkenyl-piperidinecarboxylic acid derivatives were synthesized and evaluated as gamma-aminobutyric acid uptake inhibitors. The biological assay showed that (R)-1-[4,4-bis(3-phenoxymethyl-2-thienyl)-3-butenyl] 3-piperidinecarboxylic hydrochloride possessed almost as strong GAT1 inhibitory activity as tiagabine. The synthesis and structure-activity relationships are discussed. PMID- 16246549 TI - Specific volume and compressibility of human serum albumin-polyanion complexes. AB - The ultrasound velocimetry, densitometry, and differential scanning calorimetry have been used to study the formation of the complexes between human serum albumin (HSA) and polyanions heparin (HEP) and/or dextran sulfate (DS). The values of the ultrasound velocity and specific volume allowed us to determine the specific adiabatic compressibility, phi(K)/beta(0), which reflects the degree of volume compressibility of the complexes. We showed that in the presence of HEP and DS the adiabatic compressibility of HSA decreases with increasing concentration of polyanions. HEP more strongly interacts with HSA than DS. pH of electrolyte in the range 4.7-8.5 weakly affects the adiabatic compressibility. Changes of compressibility of HSA can be caused by increase of the hydration due to the formation of the HSA-polyanion complexes and due to partial unfolding of HSA. The HSA-polyanion interaction resulted in decrease of phase transition temperature of the protein. This evidences about protein destabilization in the presence of polyanions. PMID- 16246550 TI - PET imaging and optical imaging with D-luciferin [11C]methyl ester and D luciferin [11C]methyl ether of luciferase gene expression in tumor xenografts of living mice. AB - New carbon-11 labeled D-luciferin analogs D-luciferin [(11)C]methyl ester ([(11)C]LMEster, [(11)C]1) and D-luciferin [(11)C]methyl ether ([(11)C]LMEther, [(11)C]2) were synthesized in 25-55% radiochemical yield. PET studies with [(11)C]LMEster and [(11)C]LMEther demonstrate a lower retention of the C-11 label at 45 min post-injection in luciferase expression tumor. Optical imaging with unlabeled substrate D-luciferin and radiotracers [(11)C]LMEster and [(11)C]LMEther gave tumor luciferase images within a few minutes of photon counting. PMID- 16246551 TI - N-(4-{[4-(1H-Benzoimidazol-2-yl)-arylamino]-methyl}-phenyl)-benzamide derivatives as small molecule heparanase inhibitors. AB - A novel class of N-(4-{[4-(1H-benzoimidazol-2-yl)-arylamino]-methyl}-phenyl) benzamides are described as inhibitors of the endo-beta-glucuronidase heparanase. Among them are N-(4-{[4-(1H-benzoimidazol-2-yl)-phenylamino]-methyl}-phenyl)-3 bromo-4-methoxy-benzamide (15h), and N-(4-{[5-(1H-benzoimidazol-2-yl)-pyridin-2 ylamino]-methyl}- phenyl)-3-bromo-4-methoxy-benzamide (23) which displayed good heparanase inhibitory activity (IC(50) 0.23-0.29 microM), with the latter showing oral exposure in mice. PMID- 16246552 TI - Novel histamine H3 receptor antagonists based on the 4-[(1H-imidazol-4 yl)methyl]piperidine scaffold. AB - We report the discovery of novel histamine H(3) receptor antagonists based on 4 [(1H-imidazol-4-yl)methyl]piperidine. The most potent compounds in the series (e.g., 7) result from the attachment of a substituted aniline amide to the main pharmacophore piperidine via a two-methylene linker. PMID- 16246553 TI - Peptide inhibitors of Dengue virus NS3 protease. Part 1: Warhead. AB - Substrate-based tetrapeptide inhibitors with various warheads were designed, synthesized, and evaluated against the Dengue virus NS3 protease. Effective inhibition was achieved by peptide inhibitors with electrophilic warheads such as aldehyde, trifluoromethyl ketone, and boronic acid. A boronic acid has the highest affinity, exhibiting a K(i) of 43 nM. PMID- 16246554 TI - Hexylitaconic acid: a new inhibitor of p53-HDM2 interaction isolated from a marine-derived fungus, Arthrinium sp. AB - A new inhibitor of p53-HDM2 interaction was isolated from a culture of marine derived fungus, Arthrinium sp. The structure was identified to be (-) hexylitaconic acid (1) by spectroscopic analysis. The inhibition of p53-HDM2 binding was tested by the ELISA method, and 1 inhibited the binding with an IC(50) value of 50 microg/mL. Although a number of synthetic inhibitors of p53 HDM2 interaction have been reported so far, 1 is the second inhibitor isolated from natural resources. PMID- 16246555 TI - Pentacyclic triterpenes. Part 2: Synthesis and biological evaluation of maslinic acid derivatives as glycogen phosphorylase inhibitors. AB - The synthesis of a series of maslinic acid derivatives is described and their effect on rabbit muscle glycogen phosphorylase a evaluated. Within this series of compounds, 15 (IC(50)=7 microM) is the most potent GPa inhibitor. SAR of the maslinic acid derivatives are discussed. PMID- 16246557 TI - Discovery of N-(2-aryl-cyclohexyl) substituted spiropiperidines as a novel class of GlyT1 inhibitors. AB - Screening of the Roche compound library led to the identification of cis-N-(2 phenyl-cyclohexyl)-spiropiperidine 1 as structurally novel GlyT1 inhibitor. The SAR, which was developed in this series, resulted in the discovery of highly potent compounds displaying excellent selectivity against the GlyT2 isoform. PMID- 16246556 TI - 4-hydroxyisoleucine an unusual amino acid as antidyslipidemic and antihyperglycemic agent. AB - Trigonella foenum-graecum, commonly known as fenugreek, is an annual herbaceous plant. From the seeds of T. foenum-graecum an unusual amino acid, 4 hydroxyisoleucine 5, has been isolated, which significantly decreased the plasma triglyceride levels by 33% (P<0.002), total cholesterol (TC) by 22% (P<0.02), and free fatty acids by 14%, accompanied by an increase in HDL-C/TC ratio by 39% in the dyslipidemic hamster model. PMID- 16246558 TI - Biphenyl-4-ylcarbamoyl thiophene carboxylic acids as potent DHODH inhibitors. AB - A previously discovered DHODH inhibitor series was further improved by replacing the cyclopentene ring by aromatic heterocycles. Different isomers of these compounds were prepared by the directed ortho-metallation procedure. The compounds are more active than the corresponding cyclopentene analogs and show potent effects on PBMC's proliferation. PMID- 16246559 TI - Synthesis and antibacterial activity of 9-substituted minocycline derivatives. AB - A number of 9-acylamino and 9-sulfonylamino derivatives of minocycline have been synthesized for structure-activity relationship studies. These compounds showed activity against both tetracycline-susceptible and tetracycline-resistant strains. Many of the 9-sulfonylamino derivatives exhibited improved antibacterial activity against a number of tetracycline- and minocycline-resistant Gram positive pathogens. PMID- 16246560 TI - 1-[4-(1H-Benzoimidazol-2-yl)-phenyl]-3-[4-(1H-benzoimidazol-2-yl)-phenyl]-urea derivatives as small molecule heparanase inhibitors. AB - A novel class of 1-[4-(1H-benzoimidazol-2-yl)-phenyl]-3-[4-(1H-benzoimidazol-2 yl)-phenyl]-ureas are described as potent inhibitors of heparanase. Among them are 1,3-bis-[4-(1H-benzoimidazol-2-yl)-phenyl]-urea (7a) and 1,3-bis-[4-(5,6 dimethyl-1H-benzoimidazol-2-yl)-phenyl]-urea (7d), which displayed good heparanase inhibitory activity (IC(50) 0.075-0.27 microM). Compound 7a showed good efficacy in a B16 metastasis model. PMID- 16246561 TI - Discovery of N-(2-hydroxy-2-aryl-cyclohexyl) substituted spiropiperidines as GlyT1 antagonists with improved pharmacological profile. AB - During SAR exploration of N-(2-aryl-cyclohexyl) substituted spiropiperidine as GlyT1 inhibitors, it was found that introduction of an hydroxy group in position 2 of the cyclohexyl residue considerably improves the pharmacological profile. In particular, reduction of the binding affinity at the nociceptin/orphanin FQ peptide and the mu opioid receptors was achieved. PMID- 16246562 TI - Development and evaluation of a pharmacophore model for inhibitors of aldosterone synthase (CYP11B2). AB - Recently, we proposed inhibition of aldosterone synthase (CYP11B2) as a novel strategy for the treatment of congestive heart failure and myocardial fibrosis and synthesized a large number of inhibitors. In this work, a pharmacophore model for CYP11B2 inhibitors was developed by superimposition of active and non-active compounds. This model was confirmed by the synthesis of two pyridyl substituted acenaphthene derivatives (A,B). This new class of compounds as well as the pharmacophore could be helpful for the discovery of novel inhibitors. PMID- 16246564 TI - Biological variables and prognosis of DCIS. AB - Based on current knowledge, biological factors that have been investigated in ductal carcinoma in situ (DCIS) include histology of these lesions, the impact of margin status on local recurrence, and several genetic alterations. Optimal integration of these factors in guiding optimal therapy is of great importance, since the incidence of DCIS is rising as a result of population-based mammographic screening. Mastectomy will almost always cure patients with DCIS but represents overtreatment for many. Less extensive treatment options should combine an optimal cosmetic result with the same safety for outcome of disease as mastectomy. To guide such optimal treatment, histological classification is not sufficient and additional biological factors are being investigated for their ability to predict outcome for individual patients with DCIS. In this review, the histological classification of DCIS is described and in addition the emerging knowledge on genetic alterations is summarised. For clinical management of DCIS patients, genetic or other biological factors should be identified that can predict the risk of progression of DCIS to invasive breast cancer and distant metastases. At present, insufficient knowledge on prognostic and predictive factors in DCIS is available. Research in this area is hampered by the difficulties in obtaining DCIS tumour tissue, as the tumour cells grow in the lumen of pre-existing ducts and lobules. As the recurrence rates are relatively low and the most relevant clinical endpoint, distant metastases, is indeed very rare, large numbers of patients (hundreds to a few thousand) need to be studied. Integration of translational studies into clinical trials aimed at optimising the treatment of DCIS are required to achieve this goal. PMID- 16246563 TI - Peptide inhibitors of dengue virus NS3 protease. Part 2: SAR study of tetrapeptide aldehyde inhibitors. AB - With the aim of discovering potent and selective dengue NS3 protease inhibitors, we systematically synthesized and evaluated a series of tetrapeptide aldehydes based on lead aldehyde 1 (Bz-Nle-Lys-Arg-Arg-H, K(i)=5.8 microM). In general, we observe that interactions of P(2) side chain are more important than P(1) followed by P(3) and P(4). Tripeptide and dipeptide aldehyde inhibitors also show low micromolar activity. Additionally, an effective non-basic, uncharged replacement of P(1) Arg is identified. PMID- 16246565 TI - The presence of polyglucosan bodies in temporal lobe epilepsy: its role and significance. AB - RATIONALE: Mesial temporal sclerosis (MTS) is an important pathology in temporal lobe epilepsy (TLE) and often associated with good surgical outcome, however prognostic factors for surgical outcome are conflicting. The authors examine seizure outcome after surgery for TLE due to MTS, with focus on the presence of polyglucosan bodies (PGBs), and its relation to the epileptogenic process. METHODS: Records of 44 consecutive patients with intractable TLE who underwent anterior temporal lobectomy (ATL) at JFK medical center between 1994-2001 were reviewed. Details of clinical, electrophysiological evaluation, and outcome were collected. All slides were reviewed for the presence of MTS. In twenty-one patients with MTS, detailed histology of surgical specimens was reviewed and relative concentration of PGBs was assessed. The postoperative follow-up duration ranges between 28-74 months. Surgical outcome was evaluated according to Engel's classification. RESULTS: Data of fourteen females (mean age 32.8 years) and 7 males (mean age 34.2) were analyzed. PBGs were identified in 10 patients (8 females and 2 males). All 10 patients had good surgical outcome, with 6 patients (60%) being seizure free (class 1) and 4 patients as class 2. While 11 patients (6 males and 5 females) with absent PGB, 72.8% had good surgical outcome. Of these, 7 (63.8%) had class 1, 1 patient had class 2, and 3 patients had class 3&4. Assessing several risk factors, the only significant difference between the two groups was epilepsy duration, which was significantly longer in patients with PGBs (P = 0.011). CONCLUSION: The histological presence of PGB is associated with long duration of epilepsy and could be the consequence of epilepsy. Despite prolonged duration of epilepsy, the surgical outcome in PGBs +ve is similar to PGBs -ve patients. PMID- 16246566 TI - Herpes encephalitis preceded by ipsilateral vestibular neuronitis. AB - A 74-year-old woman developed vertigo and jerk nystagmus to the left with normal cerebral imaging. Three days later she developed fever, altered mental state and left medial temporal lobe hypodensity, confirmed on lumbar puncture to be due to herpes simplex type 1 encephalitis. We propose that the patient had vestibular neuronitis caused by HSV-1 that progressed to ipsilateral temporal lobe encephalitis. PMID- 16246567 TI - Galanin attenuates beta-amyloid (Abeta) toxicity in rat cholinergic basal forebrain neurons. AB - In brains of Alzheimer's disease (AD) patients, expression of the neuropeptide galanin is significantly upregulated and galanin-immunoreactive fibers hypertrophy and hyperinnervate cholinergic neurons of the basal forebrain. However, the role of galanin in AD, whether it is detrimental or neuroprotective, remains controversial. In this study, using primary cultured neurons from the rat basal forebrain, we show that pretreatment with galanin protects cholinergic neurons against beta-amyloid-induced apoptotic cell death as judged by visual observation, MTT assay, Live/dead cell assay, TUNEL and cleaved caspase-3 staining. These effects are mimicked by the galanin receptor 2 (GALR2) agonist, AR-M1896. Western blot analysis revealed Abeta-induced decrease in phospho-PKC and phospho-Akt levels was reversed by galanin. Galanin also attenuated cleavage of caspases-3 and -9 following exposure to Abeta. These findings support a neuroprotective role for galanin and may have implications for development of compounds based on this peptide to treat AD. PMID- 16246568 TI - Treatment of older patients with AML. AB - Undertreatment of the older patients with AML can explain, in part, their inferior outcome when compared with that in younger patients. In analogy to the benefit of patients under the age of 60 years from high-dose AraC there are dosage related therapeutic effects in the patients over 60 years in particular for daunorubicin in the induction treatment, and for maintenance versus no maintenance in the post-remission treatment. Utilizing these effects can partly overcome the mostly unfavorable disease biology in older age AML, whereas the role of risk factors involved is not completely understood and the concept of dose-response needs to be requestioned. We recommend an adequate dosage of 60 mg/(m2day) daunorubicin for 3 days in a combination with standard dose AraC and 6 thioguanine given for induction and consolidation and followed by a prolonged monthly maintenance chemotherapy. Further improvements in supportive care may help delivering additional anti-leukemic cytotoxicity. As a novel approach, reduced toxicity preparative regimens may open up allogeneic transplantation for older patients with AML. Other new options like MDR modulators, antibody targeted therapies and tyrosine kinase inhibitors are under clinical investigation. A questionnaire study in patients with AML showed that according to patients' self assessment intensive and prolonged treatment did not result in decreasing quality of life. This finding did not vary by age under or above 60 years. Given the actual median age in this disease being more than 60 years the adequate management of older age AML remains as the major challenge. PMID- 16246569 TI - Intracellular interleukin-1 receptor antagonist type 1 antagonizes the stimulatory effect of interleukin-1 alpha precursor on cell motility. AB - Interleukin (IL)-1alpha, a proinflammatory cytokine, is produced as a 33 kDa protein precursor (preIL-1alpha) which is cleaved to generate the 17 kDa C terminal mature IL-1alpha (mIL-1alpha) and the 16kDa N-terminal IL-1alpha propiece (NIL-1alpha). The biological effect of IL-1alpha is regulated by the IL 1 receptor antagonist (IL-1Ra), its naturally occurring inhibitor. Four different isoforms of the IL-1Ra have been described, one secreted (sIL-1Ra) and three intracellular (icIL-1Ra1, 2, 3). Whether the icIL-1Ra1 isoform can antagonize some of the biological effects of intracellular IL-1alpha is still unknown. The aim of this study is to investigate effects of preIL-1alpha and icIL-1Ra1 on cell motility in stably transfected ECV304 cells. We show that expression of preIL 1alpha in ECV304 cells significantly increases cell motility. Furthermore, transfection with NIL-1alpha propiece also increases cell motility whereas this stimulatory effect was not observed by addition of exogenous mIL-1alpha, suggesting an intracellular effect of preIL-1alpha mediated by NIL-1alpha propiece. Co-transfection of ECV304 cells with icIL-1Ra1 completely antagonizes the stimulatory effect of preIL-1alpha and NIL-1alpha propiece on cell motility. In conclusion, NIL-1alpha propiece increases ECV304 cell motility and icIL-1Ra1 exerts intracellular functions regulating this stimulatory effect. PMID- 16246570 TI - Association between peripheral IFN-gamma producing CD8+ T-cells and disability score in relapsing-remitting multiple sclerosis. AB - A large body of evidence supports the involvement of the immune system in the pathogenesis of multiple sclerosis (MS). Nevertheless, how the peripheral T-cells phenotypes are associated with factors such as the disability score, the effects of immunomodulatory treatments, or the activation period is poorly understood. In this study, we have centered our attention on the presence of IFN-gamma and IL-4 producing CD4+ and CD8+ T-cells in the peripheral blood of 58 relapsing-remitting MS (RRMS) patients, 48 that were stable and 10 who were in relapse period, and 30 healthy controls (HC). Our results support the existence of an independent association between the percentage of IFN-gamma producing CD8+ lymphocytes and the increased levels of disability score. Furthermore, the number of IFN-gamma producing CD8+ lymphocytes and the disability score were not correlated in patients treated with interferon-beta, evidence of its possible benefits in combating a pro-inflammatory profile. Finally, we compared the T-cell populations in RRMS patients in the stable or active period, and we found a significant decrease of IFN-gamma producing CD4+ lymphocytes in active patients. In conclusion our study supports the hypothesis that different peripheral blood T cell phenotypes are associated with disability score or active period of the disease. PMID- 16246571 TI - Reactivity of zinc finger cysteines: chemical modifications within labile zinc fingers in estrogen receptor. AB - Estrogen receptor (ER, alpha isoform) is a 67 kDa zinc finger transcription factor that plays a fundamental role in both normal reproductive gland development and breast carcinogenesis, and also represents a critical molecular target for breast cancer therapy. We are investigating the structural consequences of chemical exposures thought to modify essential zinc finger cysteine residues in human ER. The current study employs mass spectrometry to probe ER zinc finger structural changes induced by a redox-reactive vitamin K3 analog, menadione; a commonly used cysteine alkylator, iodoacetic acid; and a thiol alkylating fluorophore, monobromobimane. Although they are slower to react, the sterically bulkier reagents, monobromobimane and menadione, effectively alkylate the most susceptible ER zinc finger cysteine sulfhydryl groups. Menadione arylation results first in Michael addition of the hydroquinone followed by rapid oxidation to the corresponding quinone, evidenced by a 2 Da mass loss per cysteine residue. Mass spectrometric analysis performed under MALDI conditions reveals both hydroquinone and quinone forms of arylated menadione, whereas only the quinone product is detectable under ESI conditions. Tandem mass spectrometry of a synthetic peptide encompassing the C-terminal half of the structurally more labile second zinc finger of ER (ZnF2B) demonstrates that the two nucleophilic thiols in ZnF2B (Cys-237, Cys-240) are not chemically equivalent in their reactivity to bromobimane or menadione, consistent with their unequal positioning near basic amino acids that affect thiol pKa, thereby rendering Cys 240 more reactive than Cys-237. These findings demonstrate important differential susceptibility of ER zinc finger cysteine residues to thiol reactions. PMID- 16246572 TI - Rapid detection and identification of N-acetyl-L-cysteine thioethers using constant neutral loss and theoretical multiple reaction monitoring combined with enhanced product-ion scans on a linear ion trap mass spectrometer. AB - A sensitive and specific liquid chromatography-mass spectrometry (LC-MS) method based on the combination of constant neutral loss scans (CNL) with product ion scans was developed on a linear ion trap. The method is applicable for the detection and identification of analytes with identical chemical substructures (such as conjugates of xenobiotics formed in biological systems) which give common CNLs. A specific CNL was observed for thioethers of N-acetyl-L-cysteine (mercapturic acids, MA) by LC-MS/MS. MS and HPLC parameters were optimized with 16 MAs available as reference compounds. All of these provided a CNL of 129 Da in the negative-ion mode. To assess sensitivity, a multiple reaction monitoring (MRM) mode with 251 theoretical transitions using the CNL of 129 Da combined with a product ion scan (IDA thMRM) was compared with CNL combined with a product ion scan (IDA CNL). An information-dependent acquisition (IDA) uses a survey scan such as MRM (multiple reaction monitoring) to generate "informations" and starting a second acquisition experiment such as a product ion scan using these "informations." Th-MRM means calculated transitions and not transitions generated from an available standard in the tuning mode. The product ion spectra provide additional information on the chemical structure of the unknown analytes. All MA standards were spiked in low concentrations to rat urines and were detected with both methods with LODs ranging from 60 pmol/mL to 1.63 nmol/mL with IDA thMRM. The expected product ion spectra were observed in urine. Application of this screening method to biological samples indicated the presence of a number of MAs in urine of unexposed rats, and resulted in the identification of 1,4 dihydroxynonene mercapturic acid as one of these MAs by negative and positive product ion spectra. These results show that the developed methods have a high potential to serve as both a prescreen to detect unknown MAs and to identify these analytes in complex matrix. PMID- 16246573 TI - Hydrogen/Chlorine exchange reactions of gaseous carbanions. AB - Gas-phase reactions of three typical carbanions CH(2)NO(2)(-), CH(2)CN(-), and CH(2)S(O)CH(3)(-) with the chloromethanes CH(2)Cl(2), CHCl(3), and CCl(4), examined by tandem mass spectrometry, show a novel hydrogen/chlorine exchange reaction. For example, reaction between the nitromethyl anion CH(2)NO(2)(-) and carbon tetrachloride CCl(4) forms the ion CHClNO(2)(-). The suggested reaction mechanism involves nucleophilic attack by CH(2)NO(2)(-) at the chlorine of CCl(4) followed by proton transfer within the resulting complex [CH(2)ClNO(2) + CCl(3)( )] to form CHClNO(2)(-) and CHCl(3). Two other carbanions CH(2)CN(-) and CH(2)S(O)CH(3)(-) also undergo the novel hydrogen/chlorine exchange reactions with CCl(4) but to a much smaller extent, their higher nucleophilicities favoring competitive nucleophilic attack reactions. Proton abstraction is the exclusive pathway in the reactions of these carbanions with CHCl(3). While CH(2)CN(-) and CH(2)S(O)CH(3)(-) promote mainly proton abstraction and nucleophilic displacement in reactions with CH(2)Cl(2), CH(2)NO(2)(-) does not react. PMID- 16246575 TI - Ionic liquid matrix-induced metastable decay of peptides and oligonucleotides and stabilization of phospholipids in MALDI FTMS analyses. AB - Room-temperature ionic liquid matrices (ILMs) have recently been investigated for use in matrix-assisted laser desorption/ionization (MALDI) mass spectrometry (MS) and proven to be advantageous. Literature accounts of ILM performance for biological samples document increased sensitivity and ionization efficiency. These claims have been investigated here, and are supported for MALDI TOF applications to peptides, oligonucleotides, and phospholipids. Peptides and oligonucleotides however, do not behave in the same way when ILMs are used for MALDI FTMS. As reported here, with 3 tesla MALDI FTMS peptides and oligonucleotides fragment readily. These observations contrast with those found for MALDI time-of-flight mass spectrometry. Fragmentation is apparently slower than the time required to accelerate ions in a MALDI TOF mass spectrometer, but is readily observed by MALDI FTMS. Therefore, fragmentation of these molecules must occur on a relatively slow time scale. As trapping time is extended, increased fragmentation of peptides and oligonucleotides is seen. However, phospholipids do not fragment extensively. Furthermore, use of traditional solid matrices causes significant fragmentation for this category of compound but is suppressed by use of ILMs. PMID- 16246577 TI - The design and characteristic features of a new time-of-flight mass spectrometer with a spiral ion trajectory. AB - A new time-of-flight (TOF) mass spectrometer with a corkscrew ion trajectory was designed and constructed. The spiral trajectory was realized by using four toroidal electrostatic sectors. Each had fifteen-stories made of sixteen Matsuda plates piled up inside a cylindrical electrostatic sector. The ions passed the four toroidal electrostatic sectors sequentially and revolved along a figure eight-shaped orbit on a certain projection plane. During the multiple revolutions, each ion trajectory was shifted by 50 mm per cycle on a direction perpendicular to the projection plane, thus generating a spiral trajectory. The flight path length of one cycle was 1.308 m so that the maximum flight path length became approximately 20 m. The mass resolution, mass accuracy, and ion transmission were tested by utilizing an orthogonally coupled electron ionization source. A mass resolution of 35,000 (FWHM) for m/z greater than 300 was achieved. Even in a lower mass region, mass resolutions of more than 20,000 (FWHM) were confirmed with a doublet of (12)C(5)(1)H(5)(14)N(+) and (13)C(12)C(5)(1)H(6)(+). The mass accuracy was also improved such that it was better than 1 ppm with only one internal standard peak. An ion transmission of approximately of 100% was observed for 15 cycles. PMID- 16246576 TI - Hydrocarbon polymer analysis by external MALDI fourier transform and reflectron time of flight mass spectrometry. AB - The traditional solvent-based matrix-assisted laser desorption ionization (MALDI) preparation method has been used to analyze nonpolar polymers of various molecular weights. High resolution silver cationized oligomers with masses of up to 12 KDa were measured using 9.4 tesla Fourier transform mass spectrometry (FTMS) with an external ionization source. It was observed that when time-of flight mass spectrometry was used, the spectra of polyethylene polymers showed abundant low mass fragment ions. However, these fragments were absent from the FTMS spectra. PMID- 16246578 TI - Electrospray ionization (ESI) tandem mass spectrometric analysis of meso tetrakis(Heptafluoropropyl)porphyrin. AB - The results of an electrospray ionization (ESI(+) and ESI(-)) collision-induced fragmentation investigation of meso-tetrakis(heptafluoropropyl)porphyrin (T(F)HP) are reported. The fragmentation patterns of T(F)HP show, in both ionization modes, the formation of fragments with direct covalent meso-alkyl-to-beta linkages on elimination of HF. In addition, in the ESI(+) mode, the losses of perfluoroalkyl radicals are observed. A detailed analysis of the fragmentation patterns allows conclusions to be drawn on the identification of the fragment species. Comparable cyclized products were previously produced using radical initiated solution-phase syntheses. The results presented will help to analyze meso-perfluoroalkyl-derivatized porphyrinic macrocycles that are becoming increasingly popular. PMID- 16246579 TI - Mapping protein interfaces by a trifunctional cross-linker combined with MALDI TOF and ESI-FTICR mass spectrometry. AB - Chemical cross-linking of protein complexes has gained renewed interest in combination with mass spectrometric analysis of the reaction products as it allows a rapid mapping of protein interfaces, which is crucial for understanding protein/protein interactions. The identification of cross-linking products from the complex mixtures created after the cross-linking reaction, however, remains a daunting task. To facilitate the identification of cross-linking products, we explore the use of the commercially available biotinylated cross-linking reagent sulfo-SBED (sulfosuccinimidyl-2-[6-(biotinamido)-2-(p-azidobenzamido) hexanoamido]ethyl-1,3'-dithiopropionate). This trifunctional cross-linker possesses one amine-reactive and one photo-reactive site and, additionally, allows an affinity-based enrichment of cross-linker containing species. As a model system, we chose the Ca(2+)-dependent complex between calmodulin and its target peptide M13, which represents a part of the C-terminal sequence of the skeletal muscle myosin light chain kinase. After the cross-linking reaction, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS) and one-dimensional gel electrophoresis were employed to check for the extent of cross-linking product formation. The cross-linking reaction mixtures were subjected to tryptic in-solution digestion. Biotinylated peptides, e.g., peptides that had been modified by the cross-linker as well as cross-linked peptides, were enriched on monomeric avidin beads after several washing steps had been performed. Peptide mixtures were analyzed by MALDI-TOFMS, nano-high performance liquid chromatography (HPLC)/nano-electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (ESI-FTICRMS), and tandem MS. We demonstrate that an enrichment of cross-linker containing species allows a more efficient identification of interacting amino acid sequences in protein complexes. This strategy is expected to be especially beneficial for investigating large protein assemblies. PMID- 16246580 TI - Socioeconomic/occupational risk factors for lymphoproliferative diseases in Sweden. AB - PURPOSE: The aim of the study is to investigate associations between socioeconomic and occupational factors and lymphoproliferative (LP) diseases. METHODS: We performed a follow-up study on the economically active Swedish population, based on the Swedish Family-Cancer Database. Standardized incidence ratios and 95% confidence intervals were calculated in different social classes and occupations. RESULTS: An increased risk for LP diseases was observed for physicians, plumbers, chemical process workers, and food manufacture workers among men and shoe and leather workers, mechanics, iron and metalware workers, and launderers and dry cleaners among women. CONCLUSIONS: Results suggest that socioeconomic and occupational factors have a minor effect on risk for LP diseases. Exposure to organic solvents may increase the risk for some LP diseases, although inconsistencies of findings between subcohorts do not rule out spurious associations. PMID- 16246581 TI - Self-reported hepatitis and non-Hodgkin lymphoma: an analysis according to the time since hepatitis diagnosis. AB - PURPOSE: Using data from the Selected Cancers Study, the authors tested whether the association between hepatitis infection and non-Hodgkin lymphoma (NHL) varied according to the time since the diagnosis of hepatitis. METHODS: Cases were 1177 men ages 32-60 diagnosed with NHL between 1984 and 1988. Controls were men with no history of NHL who were frequency-matched to lymphoma cases by age and cancer registry (n = 1852). Interviews were conducted to collect data on risk factors including history of hepatitis. No information on types of hepatitis was obtained. RESULTS: Logistic regression analyses indicated that history of hepatitis diagnosed 3 or more years prior to the reference date was not a significant risk factor for NHL (OR [odds ratio] = 0.92, 95% CI: 0.63-1.35). In contrast, men with a history of hepatitis diagnosed within 3 years prior to the diagnosis date had more than a five-fold increased risk of NHL (OR = 5.77, 95% CI: 1.99-16.74). CONCLUSIONS: Although the study was limited by lack of information on different types of hepatitis, the increased OR for hepatitis diagnosed more closely to the NHL diagnosis suggests that the reported association between hepatitis and NHL might partly result from increased detection of NHL in patients with recently reported hepatitis virus infection. PMID- 16246582 TI - Effects of a new Helicobacter pylori infection on height and weight in Colombian children. AB - PURPOSE: The aim of the study is to investigate whether a new infection caused by Helicobacter pylori in preschool children transiently or permanently affects height and weight. METHODS: A cohort of 347 children from three day care centers was followed up for a median of 494 days. Breath tests and anthropometric measurements were performed every 2 to 4 months. The lag effect of a new infection on linear growth during a period of 8 months was analyzed by using mixed-effects models. RESULTS: One hundred five children (30.3%) became infected during the follow-up period and accumulated 92 person-years of follow-up. A significant decrease in growth velocity was observed during the first 4 months after infection. There was no height catch-up in infected children, and after 8 months, an infected child had a cumulative difference of 0.24 cm (growth velocity; 95% confidence interval, 0.22-0.26) compared with an uninfected child. Newly infected children experienced a small decrease in weight at the first visit compared with uninfected children, which became nonsignificant after the second visit without catch-up. CONCLUSIONS: This study shows a significant and nontransient effect of infection caused by H. pylori on height and weight. Potential interventions that target infected preschool children are likely to prevent growth retardation. PMID- 16246583 TI - Is social capital a protective factor against obesity and diabetes? Findings from an exploratory study. AB - PURPOSE: The aim of this exploratory study is to examine the relationship between social capital and the important public health variables of obesity and diabetes in the adult population in the United States. METHODS: State-level correlational analyses were conducted. Predictor variables included social capital (Putnam's measure), income inequality, and poverty. Outcome variables were state-level rates of obesity and diabetes as reported by the Centers for Disease Control and Prevention Behavioral Risk Factor Surveillance System. RESULTS: Social capital and poverty had significant bivariate relationships with obesity and diabetes; multivariate linear regression showed social capital to be the strongest predictor of both outcome variables (explaining 10% of the variance in obesity and 44% of the variance in diabetes). Social capital was related inversely to both variables, indicating a protective effect. Sensitivity analyses showed that an alternate measure of income inequality had a bivariate relationship with diabetes, but was not as strong a predictor as social capital in multivariate analyses. CONCLUSIONS: These exploratory study results suggest that greater levels of social capital are protective against obesity and diabetes. This initial finding warrants subsequent empirical investigations designed to identify strategies that can be used to foster the creation of social capital in areas of the United States with an especially high prevalence of obesity and diabetes. In addition, these investigations should operate at multiple levels, examining individual, social-network, and community-level measures of social capital. PMID- 16246584 TI - Platelet counts differ by sex, ethnicity, and age in the United States. AB - PURPOSE: The aim of the study is to show that differences in platelet counts by ethnicity, sex, and age are not explained by environmental factors. METHODS: This is a cross-sectional population-based study of participants in the Third National Health and Nutrition Examination Survey. Our analytic sample included 12,142 participants, of whom 65% were women, 27% were non-Hispanic blacks, and 27% were Mexican Americans. We report weighted geometric mean platelet counts stratified by ethnicity, sex, and age and controlled for indicators of nutritional deficiencies and inflammation. RESULTS: The lowest mean platelet counts were in whites (260x10(3)/microL; 95% confidence interval [CI], 256-264), and the highest were in non-Hispanic blacks (281x10(3)/microL [95% CI, 276-286]). Older men and women of each ethnicity consistently had lower platelet counts than young adults; 60- to 69-year-olds had mean counts 7x10(3)/microL lower (p<0.001) and 70- to 90 year-olds had counts 18x10(3)/microL lower (p<0.001). Even controlling for iron deficiency, women had higher platelet counts than men (275x10(3)/microL; 95% CI, 271-279) versus 256x10(3)/microL (95% CI, 251-260; p<0.001). CONCLUSIONS: Platelet count differences by sex, ethnicity, and age are not explained by environmental covariates known to influence platelet count. PMID- 16246585 TI - Cloning and expression of TNF-alpha, IL-1beta and COX-2 in an anadromous and landlocked strain of Atlantic salmon (Salmo salar L.) during the smolting period. AB - The parr-smolt transformation involves complex modulation of immune parameters, affecting both cell populations and humoral factors. The expression of cytokines was studied in salmon cells and tissues during this period using an anadromous and a landlocked freshwater resident dwarf strain of Atlantic salmon (Salmo salar L.). The constitutive activity of three immunoregulatory genes encoding the cytokines tumour necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL 1beta) and the cyclo-oxygenase (COX) isoform COX-2 was investigated in head kidney, spleen and gill tissue from healthy, unvaccinated fish by real-time PCR. The TNF-alpha gene was generally lower expressed than COX-2 and IL-1beta1, which were approximately expressed at equal levels and constitutive expression was seen for COX-2 and IL-1beta1 in all tissues examined and at all sampling dates. The expression of all three genes in head kidney and spleen tissue seemed to be highest at the sampling in May for both strains around the time of seawater transfer suggesting an influence of smolting related hormones on cytokine expression. The gill tissue experienced the highest expression of IL-1beta1 and COX-2 at all sampling dates indicating that this organ is immunologically important. PMID- 16246586 TI - Three-dimensional whole-head optical tomography of passive motor evoked responses in the neonate. AB - Optical tomography has been used to reconstruct three-dimensional images of the entire neonatal head during motor evoked responses. Data were successfully acquired during passive movement of each arm on four out of six infants examined, from which eight sets of bilateral images of hemodynamic parameters were reconstructed. Six out of the eight images showed the largest change in total hemoglobin in the region of the contralateral motor cortex. The mean distance between the peak response in the image and the estimated position of the contralateral motor cortex was 10.8 mm. These results suggest that optical tomography may provide an appropriate technique for non-invasive cot-side imaging of brain function. PMID- 16246587 TI - Neuronal chronometry of target detection: fusion of hemodynamic and event-related potential data. AB - Event-related potential (ERP) studies of the brain's response to infrequent, target (oddball) stimuli elicit a sequence of physiological events, the most prominent and well studied being a complex, the P300 (or P3) peaking approximately 300 ms post-stimulus for simple stimuli and slightly later for more complex stimuli. Localization of the neural generators of the human oddball response remains challenging due to the lack of a single imaging technique with good spatial and temporal resolution. Here, we use independent component analyses to fuse ERP and fMRI modalities in order to examine the dynamics of the auditory oddball response with high spatiotemporal resolution across the entire brain. Initial activations in auditory and motor planning regions are followed by auditory association cortex and motor execution regions. The P3 response is associated with brainstem, temporal lobe, and medial frontal activity and finally a late temporal lobe "evaluative" response. We show that fusing imaging modalities with different advantages can provide new information about the brain. PMID- 16246588 TI - Time-frequency analysis of target detection reveals an early interface between bottom-up and top-down processes in the gamma-band. AB - The early visual gamma-band response is an oscillatory signal evoked approximately 100 ms after stimulation. While some studies have found effects of various cognitive processes on this signal, such effects could not be replicated in other studies. Accordingly, some authors have claimed that evoked gamma-band activity reflects merely sensory functions. To resolve these conflicting positions, we conducted a target detection experiment in which the feature that defined the target could be distributed over a large or a small part of the entire stimulus. Only targets covering a larger area of the entire stimulus evoked stronger gamma-band activity than standards although the over-all stimulus size was identical for all stimuli. This increase in evoked activity resulted from stronger oscillatory power and not exclusively from stronger phase-locking. In contrast, N1 and P3 amplitudes were larger for target stimuli irrespective of the distribution of the relevant stimulus feature. These results are consistent with the notion that early gamma-band activity is generated by feature-selective neural assemblies the activity of which can in fact be modulated by top-down processes. This interaction, however, may be only detectable in scalp-recorded EEG if it affects a sufficient number of neural assemblies. PMID- 16246589 TI - A distributed parameter model of cerebral blood-tissue exchange with account of capillary transit time distribution. AB - Quantitative estimates of physiological parameters associated with cerebral blood flow can be derived from the analysis of dynamic contrast-enhanced (DCE) images, using an appropriate model of the underlying tissue impulse residue function. The theoretical formulation of a distributed parameter model of tissue microcirculation, which accounts for the effects of capillary permeability and transit time distribution, is presented here. This model considers a statistical distribution of capillary-tissue units, each described by a distributed parameter model that accounts for convective transport within the capillary and transcapillary axial diffusion. Monte Carlo simulations were performed to study the confidence of the parameter estimates, and the model was used to analyze DCE CT images of patient study cases with metastatic cerebral tumors. The tumors were found to yield significantly higher estimates than normal tissues for the parameters associated with the extravasation of tracer and for the standard deviation of capillary transit times. The proposed model can be used with DCE imaging to study the microcirculatory characteristics of cerebral tumors. PMID- 16246590 TI - Identification of large-scale networks in the brain using fMRI. AB - Cognition is thought to result from interactions within large-scale networks of brain regions. Here, we propose a method to identify these large-scale networks using functional magnetic resonance imaging (fMRI). Regions belonging to such networks are defined as sets of strongly interacting regions, each of which showing a homogeneous temporal activity. Our method of large-scale network identification (LSNI) proceeds by first detecting functionally homogeneous regions. The networks of functional interconnections are then found by comparing the correlations among these regions against a model of the correlations in the noise. To test the LSNI method, we first evaluated its specificity and sensitivity on synthetic data sets. Then, the method was applied to four real data sets with a block-designed motor task. The LSNI method correctly recovered the regions whose temporal activity was locked to the stimulus. In addition, it detected two other main networks highly reproducible across subjects, whose activity was dominated by slow fluctuations (0-0.1 Hz). One was located in medial and dorsal regions, and mostly overlapped the "default" network of the brain at rest [Greicius, M.D., Krasnow, B., Reiss, A.L., Menon, V., 2003. Functional connectivity in the resting brain: a network analysis of the default mode hypothesis. Proceedings of the National Academy of Sciences of the U.S.A. 100, 253-258]; the other was composed of lateral frontal and posterior parietal regions. The LSNI method we propose allows to detect in an exploratory and systematic way all the regions and large-scale networks activated in the working brain. PMID- 16246592 TI - The selection of intended actions and the observation of others' actions: a time resolved fMRI study. AB - Whenever we plan, imagine, or observe an action, the motor systems that would be involved in preparing and executing that action are similarly engaged. The way in which such common motor activation is formed, however, is likely to differ depending on whether it arises from our own intentional selection of action or from the observation of another's action. In this study, we use time-resolved event-related functional MRI to tease apart neural processes specifically related to the processing of observed actions, the selection of our own intended actions, the preparation for movement, and motor response execution. Participants observed a finger gesture movement or a cue indicating they should select their own finger gesture to perform, followed by a 5-s delay period; participants then performed the observed or self-selected action. During the preparation and readiness for action, prior to initiation, we found activation in a common network of higher motor areas, including dorsal and ventral premotor areas and the pre supplementary motor area (pre-SMA); the more caudal SMA showed greater activation during movement execution. Importantly, the route to this common motor activation differed depending on whether participants freely selected the actions to perform or whether they observed the actions performed by another person. Observation of action specifically involved activation of inferior and superior parietal regions, reflecting involvement of the dorsal visual pathway in visuomotor processing required for planning the action. In contrast, the selection of action specifically involved the dorsal lateral prefrontal and anterior cingulate cortex, reflecting the role of these prefrontal areas in attentional selection and guiding the selection of responses. PMID- 16246591 TI - Dissociation between melodic and rhythmic processing during piano performance from musical scores. AB - When performing or perceiving music, we experience the melodic (spatial) and rhythmic aspects as a unified whole. Moreover, the motor program theory stipulates that the relative timing and the serial order of the movement are invariant features of a motor program. Still, clinical and psychophysical observations suggest independent processing of these two aspects, in both production and perception. Here, we used functional magnetic resonance imaging to dissociate between brain areas processing the melodic and the rhythmic aspects during piano playing from musical scores. This behavior requires that the pianist decodes two types of information from the score in order to produce the desired piece of music. The spatial location of a note head determines which piano key to strike, and the various features of the note, such as the stem and flags determine the timing of each key stroke. We found that the medial occipital lobe, the superior temporal lobe, the rostral cingulate cortex, the putamen and the cerebellum process the melodic information, whereas the lateral occipital and the inferior temporal cortex, the left supramarginal gyrus, the left inferior and ventral frontal gyri, the caudate nucleus, and the cerebellum process the rhythmic information. Thus, we suggest a dissociate involvement of the dorsal visual stream in the spatial pitch processing and the ventral visual stream in temporal movement preparation. We propose that this dissociate organization may be important for fast learning and flexibility in motor control. PMID- 16246593 TI - Manganese-enhanced magnetic resonance imaging of mossy fiber plasticity in vivo. AB - Mn(2+)-enhanced magnetic resonance imaging (MEMRI) was used to characterize activity-dependent plasticity in the mossy fiber pathway after intraperitoneal kainic acid (KA) injection. Enhancement of the MEMRI signal in the dentate gyrus and the CA3 subregion of the hippocampus was evident 3 to 5 days after injection of MnCl(2) into the entorhinal cortex both in control and KA-injected rats. In volume-rendered three-dimensional reconstructions, Mn(2+)-induced signal enhancement revealed the extent of the mossy fiber pathway throughout the septotemporal axis of the dentate gyrus. An increase in the number of Mn(2+) enhanced pixels in the dentate gyrus and CA3 subfield of rats with KA injection correlated (P < 0.05) with histologically verified mossy fiber sprouting. These data demonstrate that MEMRI can be used to detect specific changes at the cellular level during activity-dependent plasticity in vivo. The present findings also suggest that MEMRI signal changes can serve as an imaging marker of epileptogenesis. PMID- 16246594 TI - Category specificity in the processing of color-related and form-related words: an ERP study. AB - In this study, we investigated the spatio-temporal patterns of category-specific cortical activation elicited by the visual presentation of words whose meaning relates to a color or to a visual form or shape. We recorded the event-related potentials (ERPs) of ten healthy, right-handed volunteers while they passively read words presented tachistoscopically. As early as 150 ms after stimulus onset, the ERPs revealed significant neurophysiological differences between words and strings of hash marks. Around 200 ms after stimulus onset, we found significant differences in the ERPs elicited by color- and form-related words. We used minimum norm current estimates to investigate the spatial location of these differences. This revealed that, at the 150 ms peak, the activation advantage of words over sequences of hash marks was located in a left posterior area, proximal to what has previously been called the visual word form area. At the 200 ms peak, the advantage of words related to colors over words related to forms seemed to reside in temporal cortical areas, whereas the form-related words elicited greater activation in frontal areas than color-related words. These results provide evidence for early access to detailed category-specific representations of word meaning, with subtle differences in meaning being reflected in the activation of different cortical areas, as early as 200 ms after stimulus presentation. In line with previous studies, these differences can be related to the areas involved in the conceptual processing of sensory (visual) and action related information. PMID- 16246595 TI - Contingency awareness in human aversive conditioning involves the middle frontal gyrus. AB - In contrast to the wealth of data describing the neural mechanisms underlying classical conditioning, we know remarkably little about the mechanisms involved in acquisition of explicit contingency awareness. Subjects variably acquire contingency awareness in classical conditioning paradigms, in which they are able to describe the temporal relationship between a conditioned cue and its outcome. Previous studies have implicated the hippocampus and prefrontal cortex in the acquisition of explicit knowledge, although their specific roles remain unclear. We used functional magnetic resonance imaging to track the trial-by-trial acquisition of explicit knowledge in a concurrent trace and delay conditioning paradigm. We show that activity in bilateral middle frontal gyrus and parahippocampal gyrus correlates with the accuracy of explicit contingency awareness on each trial. In contrast, amygdala activation correlates with conditioned responses indexed by skin conductance responses (SCRs). These results demonstrate that brain regions known to be involved in other aspects of learning and memory also play a specific role, reflecting on each trial the acquisition and representation of contingency awareness. PMID- 16246596 TI - A novel, quantitative bio-assay for cholecystokinin type-1 receptor activity in the anaesthetised rat. AB - INTRODUCTION: Cholecystokinin type-1 (CCK(1)) receptors mediate many of the physiological functions of CCK including delay of gastric emptying, pancreatic enzyme secretion, intestinal motility and gallbladder contractility. Existing in vivo assays for the quantitative measurement of CCK(1) receptor mediated function are generally variable, limited in precision and require a relatively large number of animals to obtain statistically meaningful data. We found that they did not provide robust pharmacokinetic-pharmacodynamic data for profiling compounds acting at these receptors. Accordingly, here we describe a novel rat duodenal contractility assay that addresses these problems. METHODS: Rats were anaesthetised and a saline-filled balloon was inserted through the body of the stomach and secured in the duodenum approximately 1 cm from the pyloric sphincter for measurement of intra-lumenal pressure. Studies were performed to determine a dose, rate and frequency of administration of CCK8S that produced a readily quantifiable response. RESULTS: Initial experiments revealed that sustained exposure to CCK8S resulted in the rapid development of tachyphylaxis. After investigating different dosing paradigms, it was found that pulsatile delivery of CCK8S (intravenous infusion for 1 min every 10 min) produced a readily quantifiable contractile response that did not exhibit tachyphylaxis. The assay response output was defined as the number of contractions >5 mm Hg over baseline. The contractions were blocked in a dose-dependent manner by intravenous bolus injections of the CCK(1) receptor antagonists, dexloxiglumide (2 and 20 micromol/kg), and devazepide (3-100 nmol/kg) but not by the CCK(2) receptor antagonist gastrazole (10 micromol/kg). CONCLUSION: A novel, simple, high quality assay for the quantification of the in-vivo activity of CCK(1) receptor ligands is described. CCK8S delivered by pulsatile intravenous infusion to anesthetized rats produced a burst of contractile activity of the duodenum mediated by CCK(1) receptors. This activity was highly reproducible and sustained for more than 3 h providing an assay that circumvents problems associated with agonist-induced tachyphylaxis. PMID- 16246597 TI - Awareness and knowledge of epilepsy among students in a Malaysian university. AB - PURPOSE: University students represent a better-educated group of society and it is important that they possess the correct knowledge and attitude towards healthcare issues. The aim of this study was to determine the current level of awareness and knowledge of epilepsy among students at a public university in Malaysia. METHODS: A survey was conducted within the main campus of the Universiti Sains Malaysia (USM), Penang, Malaysia. Students were required to answer a series of questions on awareness and knowledge of epilepsy. The questionnaire was written in the Malay language. RESULTS: Over a one-week period, 289 students completed the self-administered questionnaire. It was found that 86.5% of students had heard or read about epilepsy, while 55.6% had observed an epileptic seizure. Only 30.7% said that they knew the cause of epilepsy and 5.3% thought epilepsy was caused by evil spirits. Epilepsy was considered hereditary by 66.9% of respondents, while 4.9% thought it was contagious. Furthermore, 60% of students did not think that epilepsy was a shameful illness. CONCLUSIONS: The findings indicate a generally favorable level of awareness and knowledge of epilepsy among students at the university. Nevertheless, there is still a need to improve certain aspects of knowledge and understanding of epilepsy among university students. PMID- 16246598 TI - Impairment of single-trial memory formation by oral methylphenidate in the rat. AB - High synaptic concentrations of dopamine and/or norepinephrine can impair the working memory function of the prefrontal cortex and impede attention and learning. Methylphenidate, a dopamine and norepinephrine transporter blocker known to facilitate these cognitive processes at low doses, was hypothesized to interfere with memory storage at doses that may raise concentrations of these neurotransmitters to systemically disruptive levels. In the present experiments, a dose of 10.0mg/kg of this drug was administered to female and male Long-Evans rats using a novel oral administration procedure designed to model the normal mode of delivery to humans. It was found to interfere with single-trial memory acquisition in a delayed object recognition test, a spontaneous learning task that involves no appetitive or aversive motivator. The time that the rats spent in overt exploration of the to-be-remembered objects during the acquisition trial was not affected, suggesting that the drug may have impaired processes of memory formation independently of interference with attention. PMID- 16246599 TI - Increased incidence of cancer in adult Gaucher disease in Western Europe. AB - The adult form of Gaucher disease (type I GD) is associated with a high prevalence of hypergammaglobulinemia and monoclonal gammopathy of undetermined significance (MGUS). A significantly increased risk of cancer, especially of hematological types, has been found in Ashkenazi-Jewish GD type 1 patients. In this study, incidence and mortality of cancer were assessed in a total of 131 GD patients of mixed ancestry in a population from Western Europe, i.e. 2 Gaucher referral centers in Germany (Dusseldorf) and the Netherlands (Amsterdam). Standardized rate ratios were determined by indirect standardization, using age- and sex-specific incidence and mortality rates of the Dutch population. A total of 14 GD patients of non-Ashkenazi-Jewish descent were identified of whom 5 had a hematologic malignancy. These numbers correspond to an increased risk of cancer of 2.5 (95% CI 1.1-4.7) and an increased risk of hematologic cancer of 12.7 (95% CI 2.6-37.0) among GD patients compared to the general population. In particular, the incidences of multiple myeloma and hepatocellular carcinoma in absence of preexisting cirrhosis were highly elevated, with standardized rate ratios of 51.1 (95% CI 6.2-184) and 141.3 (95% CI 17.1-510.5), respectively. These strongly increased risks on developing cancer suggest that measures for early detection and prevention of hematological and hepatic malignancies in patients with Gaucher type I disease are mandatory. PMID- 16246600 TI - Regional differences in the in vitro penetration of methylsalicylate through equine skin. AB - Commercial formulations of non-steroidal anti-inflammatory drugs (NSAIDs) are developed for human use but the extent to which they will pass through equine skin is unknown. Skin was harvested from five Thoroughbred geldings from the thorax, groin and leg (dorsal metacarpal) regions and frozen (-20 degrees C) until required. Two grams of methylsalicylate (MeSa) gel was applied to defrosted full-thickness samples in diffusion cells and the penetration of MeSa and its active metabolite, salicylate (Sa), through skin samples were measured over 24 h. Significantly higher (P < or = 0.02) total salicylate (AUC; MeSa + Sa) penetrated through skin from the leg region (5491.3 h mg/L), compared to thorax (3710.7 h mg/L) and groin (3571.5 h mg/L). In addition, there was a significantly higher (P0.01) rate of penetration of total Sa through leg skin in the first 6h after application. It was concluded that the commercial formulation of MeSa would achieve therapeutic levels of total salicylate beneath sites of topical application, with a faster and more pronounced response through the leg region, compared to the upper body. PMID- 16246601 TI - Animal sexual abuse: a veterinary taboo? PMID- 16246602 TI - Fighting tooth and CLAW for more monoclonal antibodies against canine leucocyte antigens. PMID- 16246603 TI - The cranial cervical ganglion and its branches in the yak (Bos grunniens). AB - The heads and necks of 10 yaks were dissected to study the shape, location, arrangement, and branches of the cranial cervical ganglion. The ganglion was a greyish fusiform structure, mean length 19.72 mm, width 7.65 mm and depth 4.55 mm, located on the rostrolateral surface of the m. longus capitis. Approximately 25% of the ganglion was covered by the tympanic bulla, the rest by the m. stylohyoideus. The branches of the cranial cervical ganglion included the internal and external carotid nerves, sympathetic trunk and the branches connecting with the glossopharyngeal, vagus and hypoglossal nerves. In one animal the right cranial cervical ganglia was a greyish pyramidal structure 10 mm long, 8 mm wide and 5 mm thick but the left ganglion was similar to those found in the other specimens examined. PMID- 16246604 TI - Interaction of Tritrichomonas foetus with the reproductive tract of experimentally infected female BALB/c mice: ultrastructural evaluation. AB - The interaction of Tritrichomonas foetus with its host is a complex process that involves colonisation, attachment and persistence. The goal of the present study was to describe the interaction of T. foetus with the genital tract using a model of non-oestrogenised female BALB/c mice which had been intravaginally infected with a suspension of T. foetus during oestrus. Animals were sacrificed after 10 weeks and the uteri fixed and processed for light and electron microscopy. Ultrastructural analysis showed that the attached protozoa interacted with the mucosa through a somal projection. With an amorphous secretion at the protozoa host cell interface. There was no direct contact between the protozoal plasma membrane and the epithelial cell membrane. Our results demonstrated the participation of an active phagocytosis and the destruction of T. foetus by eosinophils. PMID- 16246605 TI - Colic--Some bright views on the horizon. PMID- 16246606 TI - Improving claw trimming in cattle: onwards from the Dutch Technique. PMID- 16246607 TI - Spirocercosis-associated pyothorax in dogs. AB - The records of five dogs diagnosed with spirocercosis-associated pyothorax were retrospectively reviewed. On admission, the most common clinical findings were weakness, anorexia, depression, dyspnoea and fever. All dogs had typical oesophageal Spirocerca lupi granulomas. Contrast thoracic radiography revealed contrast material leakage from the oesophagus into the pleural cavity in two dogs only. All dogs were medically treated with broad-spectrum antibiotics, repeated chest tube drainage and thoracic lavage, and four of the dogs were treated with doramectin. One dog was euthanased due to a concurrent non-related disease at the owners request while the remaining four survived. S. lupi infection should be included in the differential diagnoses of canine pyothorax in endemic areas. Conservative medical management of pyothorax with antihelmintic medication was shown to be effective in the treatment of S. lupi-associated pyothorax. PMID- 16246608 TI - Indicators of brain mass-induced herniation. PMID- 16246609 TI - Endoscopic examination of the upper respiratory tract and oesophagus in small ruminants: technique and normal appearance. AB - The aim of this study was to describe the technique of rhinolaryngotracheobronchoscopy and oesophagoscopy in small ruminants and to report the normal endoscopic appearance of the structures of the upper respiratory tract and oesophagus. Thirty sheep and 30 goats, all clinically normal, underwent rhinolaryngotracheobronchoscopy and oesophagoscopy using a flexible endoscope (length 100 cm; diameter 4 mm). The anatomical features of the structures and the appearance of the mucosa were investigated. The degree of mucosal pigmentation varied between breeds. The appearance of the pharyngeal tonsil and the shape of the trachea in cross-section showed most individual variation. The technique was easily performed and sedation was necessary in only a few cases. No major complications were encountered. It was concluded that the procedure facilitated more accurate diagnosis and prognosis of conditions of the upper respiratory tract and oesophagus. PMID- 16246610 TI - Moving ahead in language: observations on a report of precocious language development in 3-4 year old children with spinal muscular atrophy type II. AB - Benony and Benony in a recent issue of this Journal have presented new data on the precocity of language acquisition in children with type II spinal muscular atrophy (SMA), at age 36-47 months. They refer to our interim report of advanced early language development in these motor-impaired children, which covers the age period 18-35 months. Here, we provide more details of our findings and discuss them in relation to their report and our theory of the role of the procedural system in language learning. PMID- 16246612 TI - Molecular cloning and expression pattern of 14 kDa apolipoprotein in orange spotted grouper, Epinephelus coioides. AB - A novel fish-specific apolipoprotein (apo-14 kDa) has been recently cloned from eel and pufferfish. However, its expression pattern has not been elucidated. In this study, EcApo-14 has been screened from hypothalamic cDNA library of male orange-spotted grouper, which shows 62.9%, 51%, 46.9%, 43.2%, and 31.9% identities to Apo-14 of European flounder, pufferfish, Japanese eel, gibel carp, and grass carp, respectively. RT-PCR analysis reveals that this gene is first transcribed in neurula embryos and maintains a relatively stable expression level during the following embryogenesis. EcApo-14 transcripts are at a very high level during embryonic and early larval development in the yolk syncytial layer (YSL), and decrease in YSL and form intense staining in liver at 3 days after hatching. In adult tissues, EcApo-14 is predominantly expressed in liver and brain. The data suggested that EcApo-14 might play an important role in liver and brain morphogenesis and growth. PMID- 16246611 TI - Performance of bean bruchids Callosobruchus maculatus and Zabrotes subfasciatus (Coleoptera: Bruchidae) reared on resistant (IT81D-1045) and susceptible (Epace 10) Vigna unguiculata seeds: relationship with trypsin inhibitor and vicilin excretion. AB - Callosobruchus maculatus (Cm) and Zabrotes subfasciatus (Zs) were reared on resistant (IT81D-1045) and on susceptible (Epace 10) cowpea seeds. The emergence of adult insects, total developmental period (TDP) and excretion of trypsin inhibitor and vicilin were determined for both bruchid populations. Parameter evaluation showed that the Zs populations emerged from both seeds had no significant differences in emergence and TDP. The Cm population raised from resistant seeds had lower emergence (5.6+/-1.3%) and delayed TDP (46+/-1.25 days) than those emerged from susceptible seeds. The excretion of defense proteins showed that Zs reared in resistant seeds excreted 1.7 times more trypsin inhibitor, but this did not affect emergence or TDP. Furthermore, Cm population emerged from resistant seeds excreted 7 times higher vicilin and 0.4 times less trypsin inhibitor than that emerged from susceptible seeds. These results indicate that vicilins from resistant seeds are involved to significantly longer TDP (46 days) and also drastic reduction of insect emergence ( approximately 5%) of C. maculatus. PMID- 16246614 TI - [For ultrasound-guided embryo transfer]. PMID- 16246613 TI - [The use of GnRH analogues in early and advanced breast carcinomas]. AB - Breast cancer is often an estrogen-dependent disease. The primary goals of the treatment of breast carcinomas are multiple, depending on the situation in which the patients are treated. In adjuvant setting, the aims are to delete the time of relapse, to increase the overall survival, and to offer to the patients the best quality of life they may expect. Tamoxifen is the standard hormonal agent for premenopausal women with receptor-positive breast cancer. Recent data show an increasing role for aromatase inhibitors in postmenopausal women. In metastatic setting, the primary goals are improved quality of life and prolonged survival; effective therapies with minimal toxicity, such as endocrine therapy, are highly desirable and should be considered a primary option over chemotherapy for selected estrogen-receptor positive patients. Ovarian ablation has been worldwide used. Methods of irreversible ovarian ablation include surgical oophorectomy and ovarian irradiation. Potentially reversible castration can be medically accomplished using luteinizing hormone releasing hormone analogues (LHRH agonists). In the metastatic setting, ovarian ablation (induced by the use of LHRH agonists or by surgical ovarian ablation) and tamoxifen monotherapies produce comparable outcomes, and may be more effective when used together (combined estrogen blockade). In advanced breast cancer, the combination prolongs the progression-free survival and increases response rates and duration of response rate relative to the use of LHRH agonist alone. In the adjuvant setting, data suggest that ovarian ablation followed by tamoxifen produces similar results to those obtained with adjuvant chemotherapy in hormone-receptor positive breast cancer women. The value of combining these modalities remains unclear, but the addition of the LHRH analogue goserelin to standard treatment results in a significant benefit in terms of relapse-free and overall survival, especially for estrogen-receptor positive patients. Finally, considering the efficacy of the new aromatase inhibitors, the interest of combining these drugs with the LHRH analogues has yet to be defined, both for pre- and post-menopausal patients. PMID- 16246615 TI - Effect of ultrasonically induced cavitation on inhibition behavior of polyethylene glycol on carbon steel corrosion. AB - In this investigation the influence of 24 kHz ultrasound wave upon the corrosion of carbon steel in 3N sulphuric acid at 25 degrees C in the presence of inhibitors was studied. The inhibitors were polyethylene glycols (PEG) in different molecular weights (from 400 up to 10,000 gmol(-1)). The polarization and impedance spectroscopy results show the effectiveness of polyethylene glycols on the cavitation-corrosion inhibition of carbon steel in sulphuric acid. The inhibition efficiency is increased with increasing mean molecular weight of polymer and its concentration. The weight loss method has confirmed these results. The analysis of SEM images indicates that these inhibitors prevent propagation of pits on the eroded specimen. The inhibition effect of PEGs can be attributed to cushioning effect of adsorbed polymers on cavitation phenomenon produced by bubble collapse. PMID- 16246616 TI - Immunohistochemical analysis of P57(kip2), p53 and hsp60 expressions in premalignant and malignant oral tissues. AB - To investigate the expressions and significances of P57(kip2), p53 and hsp60 proteins in the dysplasia-carcinoma sequence of oral mucosa. A retrospective study was performed in 10 cases of normal oral mucosa, 79 cases of leukoplakia and 67 cases of squamous cell carcinoma (SCC). The expressions of P57(kip2), p53 and hsp60 proteins were detected in the tissue samples of these populations using immunohistochemical method. P57(kip2) expression was decreased in oral leukoplakia with moderate or severe dysplasia, and further decreased in oral SCC. Negative expression of P57(kip2) was significantly associated with advanced tumor size, the occurrence of lymph node metastasis and the advanced clinical stage in oral SCC. The overall 5-year survival rate in the P57(kip2) positive group was significantly higher than that in the P57(kip2) negative group. P57(kip2) expression was decreased in oral leukoplakia with moderate or severe dysplasia, and further decreased in oral SCC. It was a remarkable progressive and prognostic biomarker in oral SCC. PMID- 16246617 TI - Free osseous and soft tissue surgical margins as prognostic factors in mandibular osteosarcoma. AB - Osteosarcoma is an infrequent, locally aggressive neoplasm in the head and neck region. To date, surgery is the mainstay of treatment. However, patients with mandibular osteosarcomas usually have a locally advanced disease at diagnosis and therefore represent a therapeutic challenge because surgical margins are difficult to obtain due to aesthetic and functional concerns. To evaluate possible prognostic factors implicated in recurrence, persistence or relapse in osteosarcoma of the mandible, with special reference to the soft tissue and bone surgical margins. A series of 20 patients with mandibular osteosarcomas treated at the Instituto Nacional de Cancerologia (Mexico) from 1985 to 1999 are reviewed. There were 14 female and 6 male patients. Twelve cases were treated with surgery alone, 3 patients with surgery and adjuvant radiotherapy, 1 had neoadjuvant chemotherapy followed by surgery, 1 had neoadjuvant chemotherapy, surgery and postoperative radiotherapy, 1 with surgery and adjuvant chemotherapy, 1 with surgery followed by adjuvant chemotherapy and radiotherapy and one patient rejected treatment. Between 1985 and 1992 these neoplasms were treated by means of total mandibulectomy, independently of tumor size, but between 1993 and 1999 the policy was to practice smaller resections but long enough to obtain macroscopic surgical free margins. In the first period the relationship between mandibular size resection and tumor size was 1.9, meanwhile in the second period the relation was 1.5. There was not significant difference between both periods in terms of tumor size (6.0 cm vs. 6.02 cm at the time of surgery) nor in local control and survival. Soft tissue involvement as reported by histological study was strongly associated with recurrence (p = 0.0024). Overall 5-year survival was 20%. A policy of total mandibulectomy is not associated with a better local control or survival. Extent of resection must be tailored with tumor size. Extent of margins in soft tissue is the limiting factor for local control. PMID- 16246620 TI - Preface. PMID- 16246618 TI - Biology, biochemistry and the molecular machinery of Archaea. PMID- 16246621 TI - A randomized controlled trial of an educational intervention on Hellenic nursing staff's knowledge and attitudes on cancer pain management. AB - The purpose of this randomized controlled study was to explore the effectiveness of an educational intervention on nurses' attitudes and knowledge regarding pain management and to explore associations with nurses' characteristics. A four Solomon group experimental design was employed to assess the effect of the intervention and potential effects of pre-intervention testing. One hundred and twelve nurses were randomized to two intervention and two control groups. The intervention was based on viewing a series of educational videotapes and case scenarios. The Validated Hellenic version of the Nurses Knowledge and Attitudes Survey Regarding Pain (GV-NKASRP) was used. Pre-intervention scores revealed various limitations in regard to pain assessment and management. At the pre-test, the average number of correct answers was 17.58+/-7.58 (45.1%+/-19.3% of total questions). Pre-intervention scores differed significantly among participants with different educational backgrounds (P < 0.0001). A significant effect of pain education on total knowledge scores as well as regarding specific questions was detected. Intervention group participants provided 6.11+/-5.55 additional correct answers (15.66%+/-14.23% improvement, P < 0.0001), and they exhibited significantly improved post-test scores compared to controls (26.49+/-5.24 vs. 18.75+/-4.48; P < 0.0001). A potential negative effect of pre-test on knowledge gain for specific items and for total scores was detected. These findings suggest low pre-test knowledge scores among Hellenic oncology nurses and a significant effect of the intervention. PMID- 16246622 TI - Recent findings in immunology give tuberculosis vaccines a new boost. AB - Tuberculosis remains a major health threat, solved by neither chemotherapy nor the current vaccine, BCG. Although a new generation of vaccine candidates is ready for field trials, further improvements will be required. A successful vaccination regime must stimulate memory T cells and, at the same time, avoid exhaustion of memory and suppression by regulatory mechanisms. The most probable scenario is priming with one vaccine candidate followed by boosting with a another vaccine candidate. For clinical trials, biomarkers need to be defined with T cells alternating between lung and periphery as prime indicator cells. PMID- 16246623 TI - DNA microarray genotyping of N-acetyltransferase 2 polymorphism using carbodiimide as the linker for assessment of isoniazid hepatotoxicity. AB - The antituberculous drug isoniazid (INH) is acetylated by N-acetyltransferase 2 (NAT2), and the frequency of INH-induced hepatotoxicity is determined by the NAT2 genotype. NAT2 genotyping is not done routinely in hospital laboratories because of its difficulty. Use of microarrays for research is becoming common and its expectations of clinical application are increasing. In this study, we attempted to develop an easier method of NAT2 genotyping for clinical use. We devised a novel oligonucleotide-based DNA microarray for NAT2 genotyping using a recently developed technique for attaching oligonucleotide probes to poly carbodiimide coated glass slides, which achieves a stronger hybridization signal and better specificity than the more widely used aminosilane-coated slides. To assess the validity of this microarray, four clones with NAT2 mutations and DNA from 42 tuberculosis patients were investigated by the microarray method and by restriction fragment length polymorphism analysis. The results of genotyping by these two methods were in agreement. Analysis of the relationship between the NAT2 phenotype determined by the DNA microarray and the risk of INH-induced hepatotoxicity revealed that slow acetylators had a significantly higher risk. These findings suggest that our microarray may be clinically useful for predicting drug reactions to INH. PMID- 16246624 TI - Regulation of immune responses to Mycobacterium tuberculosis secretory antigens by dendritic cells. AB - Towards elucidating the immune responses induced by antigens from the Mycobacterium tuberculosis (M. tb) RD-1 region, we have been characterizing their interactions with dendritic cells (DCs) and their precursors. We have shown that incubation of bone marrow DC precursors with M. tb antigens induces the differentiation of DC precursors and also the maturation of various DC subsets. While MTSA differentiated DCs were immature, MTSA matured DCs were terminally mature. However, regardless of their maturation status M. tb secretory antigen activated DCs down-regulated pro-inflammatory T helper cell responses to a subsequent challenge with M. tb cell extract (CE) while increasing regulatory responses. Investigations into the underlying mechanisms showed that stimulation with M. tb CE changed the polarization of antigen-activated DCs from DC1 to DC2. This resulted in secretion of high levels of IL-10 and TGF-beta together with increased surface expression of CD86. Blocking either IL-10 or TGF-beta or CD86 restored Th1 responses to CE antigens. Conversely, treatment of antigen-activated DCs with IL-12 and/or IFN-gamma fully restored Th1 responses of CE antigens. These results indicate that M. tb strategically secretes antigens from infected macrophages to down-regulate pro-inflammatory immune responses at sites of infection. PMID- 16246625 TI - Exploring drug action on Mycobacterium tuberculosis using affymetrix oligonucleotide genechips. AB - DNA microarrays have rapidly emerged as an important tool for Mycobacterium tuberculosis research. While the microarray approach has generated valuable information, a recent survey has found a lack of correlation among the microarray data produced by different laboratories on related issues, raising a concern about the credibility of research findings. The Affymetrix oligonucleotide array has been shown to be more reliable for interrogating changes in gene expression than other platforms. However, this type of array system has not been applied to the pharmacogenomic study of M. tuberculosis. The goal here was to explore the strength of the Affymetrix array system for monitoring drug-induced gene expression in M. tuberculosis, compare with other related studies, and conduct cross-platform analysis. The genome-wide gene expression profiles of M. tuberculosis in response to drug treatments including INH (isoniazid) and ethionamide were obtained using the Affymetrix array system. Up-regulated or down regulated genes were identified through bioinformatic analysis of the microarray data derived from the hybridization of RNA samples and gene probes. Based on the Affymetrix system, our method identified all drug-induced genes reported in the original reference work as well as some other genes that have not been recognized previously under the same drug treatment. For instance, the Affymetrix system revealed that Rv2524c (fas) was induced by both INH and ethionamide under the given levels of concentration, as suggested by most of the probe sets implementing this gene sequence. This finding is contradictory to previous observations that the expression of fas is not changed by INH treatment. This example illustrates that the determination of expression change for certain genes is probe-dependent, and the appropriate use of multiple probe-set representation is an advantage with the Affymetrix system. Our data also suggest that whereas the up-regulated gene expression pattern reflects the drug's mode of action, the down-regulated pattern is largely non-specific. According to our analysis, the Affymetrix array system is a reliable tool for studying the pharmacogenomics of M. tuberculosis and lends itself well in the research and development of anti-TB drugs. PMID- 16246626 TI - Autoimmune hepatic inflammation by vaccination of mice with dendritic cells loaded with well-differentiated hepatocellular carcinoma cells and administration of interleukin-12. AB - Vaccination of mice with dendritic cells loaded with Hepa1-6, well-differentiated hepatocellular carcinoma cell line (DC/Hepa1-6), induced cytotoxic T lymphocytes against Hepa1-6. Liver-specific inflammation was generated by vaccination of mice with DC/Hepa1-6 and subsequent administration of interleukin (IL)-12. Vaccination with DCs loaded with MC38 or B16 and administration of IL-12 did not generate significant liver-specific inflammation. Splenic T cells from DC/Hepa1-6 vaccinated mice showed proliferative response by stimulation with S-100 protein of the liver and showed cytotoxic activity to hepatocytes. Hepatic mononuclear cells from DC/Hepa1-6 + IL-12-treated mice also showed cytotoxic activity to hepatocytes. Adoptive transfer of splenocytes from DC/Hepa1-6-vaccinated mice produced hepatic inflammation in recipient mice that had been pretreated with IL 12. IL-12 upregulated the expression of adhesion molecules and chemokines in the liver. In conclusion, CTLs responsive to hepatocytes induced by DC/Hepa1-6 and enhanced expression of adhesion molecules and chemokines in the liver by IL-12 would produce autoimmune hepatic inflammation. PMID- 16246627 TI - Infective endocarditis complicating hypertrophic obstructive cardiomyopathy. AB - Infective endocarditis is a rare complication of hypertrophic cardiomyopathy. It's estimated incidence is 1.4 per 1000 person/year in all patients and it increases to 3.8 per 1000 person/year in patients with left ventricular outflow obstruction. The most common site of vegetation is the ventricular aspect of anterior mitral valve leaflet. We report a case of a 43-year-old man who was admitted for mitral infective endocarditis resulting in severe mitral regurgitation complicating a hypertrophic obstructive cardiomyopathy. The patient underwent mitral valve replacement. Post-operative outcome was good with relieve of symptom and resolution of left ventricular outflow obstruction. Literature data are reviewed. PMID- 16246628 TI - Modulation of dendritic spines in epilepsy: cellular mechanisms and functional implications. AB - Epilepsy patients often suffer from significant neurological deficits, including memory impairment, behavioral problems, and psychiatric disorders. While the causes of neuropsychological dysfunction in epilepsy are multifactorial, accumulating evidence indicates that seizures themselves may directly cause brain injury. Although seizures sometimes result in neuronal death, they may also cause more subtle pathological changes in neuronal structure and function, including abnormalities in synaptic transmission. Dendritic spines receive a majority of the excitatory synaptic inputs to cortical neurons and are critically involved in synaptic plasticity and learning. Studies of human epilepsy and experimental animal models demonstrate that seizures may directly affect the morphological and functional properties of dendritic spines, suggesting that seizure-related changes in spines may represent a mechanistic basis for cognitive deficits in epilepsy. Novel therapeutic strategies directed at modulation of spine motility may prevent the detrimental effects of seizures on cognitive function in epilepsy. PMID- 16246629 TI - Seizure clustering. AB - Seizure clusters, also known as repetitive or serial seizures, occur commonly in epilepsy. Clustering implies that the occurrence of one seizure may influence the probability of a subsequent seizure; thus, the investigation of the clustering phenomenon yields insights into both specific mechanisms of seizure clustering and more general concepts of seizure occurrence. Seizure clustering has been defined clinically as a number of seizures per unit time and, statistically, as a deviation from a random distribution, or interseizure interval dependence. This review explores the pathophysiology, epidemiology, and clinical implications of clustering, as well as other periodic patterns of seizure occurrence. Risk factors for experiencing clusters and potential precipitants of clustering are also addressed. PMID- 16246630 TI - Neuroethology application for the study of human temporal lobe epilepsy: from basic to applied sciences. AB - The aim of this investigation was to apply neuroethology to the study of human temporal lobe epilepsy (TLE). For this purpose, 42 seizures in 7 patients recorded during video/EEG monitoring (1997-1998) were analyzed by means of a behavioral glossary containing all behaviors. Video recordings were reobserved, and all patients' behaviors were annotated second-by-second. Data were analyzed using Ethomatic software and displayed as flowcharts including frequency, mean duration, and sequential statistic interaction of behavioral items (chi2 > or = 10.827, P<0.001). Flowcharts of (1) a group of seizures from a single patient, (2) the sum of four seizures per patient of two patients with right and five patients with left TLE, and (3) the comparison of left versus right TLE are shown. Well-established data in the literature were confirmed, such as aura (especially epigastric), contralateral lateralization value of dystonia and version, consciousness and language alterations in ictal and postictal periods, mostly with respect to dominant hemisphere involvement, among others. Less well established data such as awakening seizures in TLE patients, lateralization value of facial wiping (ipsilateral to the focus), statistically significant associations between behavioral pairs (dyads), and new behavioral sequences in TLE were also observed. We suggest that neuroethology also has great potential in the study of human epilepsy semiology. This work had an important role in method standardization for human epilepsy, setting the basis for the development of future clinical studies including correlation with other diagnostic methods (EEG, magnetic resonance, and SPECT). The next step will be the comparative study of seizures of patients with left and right TLE, with a greater number of patients, and the development of a digital video library. PMID- 16246631 TI - Major depressive comorbidity in epilepsy as a reactive process to a chronic condition. PMID- 16246632 TI - Reversal of vision metamorphopsia: a manifestation of focal seizure due to cortical dysplasia. AB - Upside-down reversal of vision has rarely been reported in the literature. The reported causes are diverse, including posterior circulation stroke, tumors, trauma, and multiple sclerosis. The term seizure has been used in only two cases in the literature, the cause of which was stroke. To our knowledge, this is the first reported case of cortical dysplasia in the posterior parietal cortex as the cause of complex partial seizures beginning with upside-down reversal of vision. As the pathophysiological characteristics of this phenomenon remain unclear, this case implies that the posterior parietal cortex is a possible anatomical localization of the central integrator of visual extra-personal orientation. PMID- 16246634 TI - Eye movement desensitization and reprocessing in the treatment of posttraumatic stress disorder in a patient with comorbid epilepsy. AB - Whether eye movement desensitization and reprocessing (EMDR) treatment of posttraumatic stress disorder (PTSD) causes reactivation of epilepsy is as yet unclear. A 34-year-old woman was treated in an inpatient multimodal psychotherapeutic setting with EMDR for PTSD resulting from sexual harassment and for a moderate depressive episode. She had been diagnosed with idiopathic generalized absence epilepsy in childhood, but had experienced no seizures under lamotrigine medication since 1999. After the second EMDR session, clinical seizures in the form of absences occurred, and were validated by electroencephalography. The seizures ceased after medication with benzodiazepines and an increase in the lamotrigine level. She underwent four more sessions of EMDR treatment successfully without further seizures. Possible triggers are discussed, especially as to whether EMDR treatment played a role in reactivating epilepsy. Further research and publications on the application of EMDR in epilepsy patients are needed. PMID- 16246633 TI - Spatial learning deficits and emotional impairments in pentylenetetrazole-kindled rats. AB - Pentylenetetrazole (PTZ) is a chemical kindling agent used to examine the efficacy of potential anticonvulsants in rats. However, the extent to which PTZ mimics postseizure symptoms of epilepsy has not been thoroughly examined. This study assessed whether PTZ-induced seizures produce cognitive and emotional deficits that mimic those observed in many epileptic patients. Rats were given 30mg/kg PTZ or vehicle (intraperitoneally) every other day for 28 days. Those rats exhibiting consistent seizure activity were tested for learning ability and emotional reactivity, beginning 1 week following a single challenge dose of PTZ. Rats given PTZ made more reference memory errors in a radial arm water maze task, and exhibited emotional abnormalities in the forced swim test, the systematic handling test, and the open-field exploratory maze. Histological analysis revealed neuronal loss in the CA1 area and increased mossy fiber sprouting in the dentate gyrus, similar to what is observed in human epilepsy. These results indicate that PTZ kindling provides a useful model of postseizure dysfunction, which can serve as a screen for potential treatments for those cognitive, emotional, and neuropathological deficits that resemble those symptoms observed in human epilepsy. PMID- 16246635 TI - Autism and epilepsy: cause, consequence, comorbidity, or coincidence? AB - Autism is associated with epilepsy in early childhood, with evidence suggesting that individuals with both autism and more severe cognitive impairment are at higher risk. However, the incidence of an abnormal electroencephalogram and/or epilepsy in the full range of pervasive developmental disorders (PDDs) is not well defined. This naturalistic study addresses the incidence of epilepsy and electroencephalographic abnormalities in children with PDDs. The clinical history and electroencephalograms of 56 children diagnosed with PDD-not otherwise specified, autism, or Asperger syndrome were retrospectively reviewed. Forty percent of children with autism were diagnosed with epilepsy. Abnormal electroencephalograms and epilepsy occurred at significantly higher rates in children in the more impaired range of the autism spectrum (P<0.05). These findings suggest that the use of neurological investigative techniques such as electroencephalography should be a consequence of careful clinical evaluation and should be considered routinely during evaluation of more impaired individuals. PMID- 16246636 TI - Psychosocial intervention in pediatric epilepsy: a critique of the literature. AB - It is well documented that youth with epilepsy are at increased risk for psychopathology. The current literature supports a biopsychosocial model of adjustment to pediatric epilepsy, and implies that interventions focused on changing youths' cognitions and illness appraisals, as well as enhancing their coping skills, may be an effective treatment for psychosocial maladjustment associated with pediatric epilepsy. The purpose of this article is to review and critique the extant literature covering psychological interventions that target psychosocial adjustment in youth with seizures followed by those aimed at reducing seizure frequency. For health care professionals treating epilepsy, establishing evidence-based interventions that target psychiatric difficulties in youth with epilepsy should be paramount in the promotion of optimal epilepsy outcomes. Thus, future recommendations for clinical endeavors and research proposals are also presented. PMID- 16246638 TI - Genetic diversity and genetic burden in humans. AB - We discuss categories of genetic diversity in humans. Neutral diversity, population differences in frequencies of genetic markers that we think are invisible to natural selection, provides a passive record of population history but is otherwise of little interest in human biology. Genetic variation related to disease can be separated into mutational noise and variation due to selection, either ongoing selection else effects of a past environment. We distinguish consequences of genetic diversity for fitness, relevant to evolution, and consequences for well-being, relevant to medicine and public health. We call genetic variation that causes impairment of health or well-being of individual humans "apparent genetic burden" and variation that has effects on fitness but not well-being "unapparent genetic burden". We use "burden" to distinguish these notions from the classical concept of "genetic load" that refers to effects on population fitness, a concept formulated by Morton et al. [Morton, N.E., Crow, J.F., Muller, H.J., 1956. An estimate of the mutational damage in man from data on consanguineous marriages. Proc. Natl. Acad. Sci. U.S.A. 42, 855-863]. We distinguish adapted genes and adapted genotypes: an adapted gene is a gene that increases fitness of its bearer either in heterozygous or homozygous state or both, while an adapted genotype is a genotype that increases fitness of its bearer but is not transmitted intact to future generations. Balanced polymorphisms in which the heterozygote is superior in fitness may generate most adapted genotypes. In the face of major rapid environmental change adapted genotypes appear first but over time they are replaced by adapted genes. The presence of adapted genotypes is a good indication of recent environmental change: for example, there are apparently many polymorphisms in domestic animals of this nature, responses to domestication, and many fewer in wild animals (and in humans). PMID- 16246639 TI - WITHDRAWN: Impact of overexpressed secretory-pathway genes on the secretion of IFNgamma in Hansenula polymorpha applying an rDNA-cointegration approach for assessment. AB - gamma-Interferon (IFNgamma) is poorly secreted from recombinant Hansenula polymorpha cells in the form of hyperglycosylated molecules. A selection of four secretory-pathway genes was assessed for possible impact on secretion improvements. For assessment, rDNA integration vectors harbouring the H. polymorpha-derived KAR2, PDI, SSO2 or CNE1 gene were co-integrated along with the IFNgamma-integration/expression vector. Overexpression of CNE1 resulted in an increased secretion of the cytokine, predominantly consisting of molecules of distinct size. Deglycosylation with PGNaseF resulted in an M(r) reduction of the secreted IFNgamma corresponding to the removal of two N-linked glycoside chains. Coexpression of KAR2, PDI or SSO2 exhibited no effect. PMID- 16246641 TI - Gene expression and DNA repair in progeroid syndromes and human aging. AB - Human progeroid syndromes are caused by mutations in single genes accelerating some but not all features of normal aging. Most progeroid disorders are linked to defects in genome maintenance, and while it remains unknown if similar processes underlie normal and premature aging, they provide useful models for the study of aging. Altered transcription is speculated to play a causative role in aging, and is involved in the pathology of most if not all progeroid syndromes. Previous studies demonstrate that there is a similar pattern of gene expression changes in primary cells from old and Werner syndrome compared to young suggesting a presence of common cellular aging mechanisms in old and progeria. Here we review the role of transcription in progeroid syndromes and discuss the implications of similar transcription aberrations in normal and premature aging. PMID- 16246642 TI - Validation of a liquid chromatography-mass spectrometry method to assess the metabolism of bupropion in rat everted gut sacs. AB - We have developed a rapid, sensitive and selective LC-MS method for the simultaneous assay of bupropion and its metabolite hydroxybupropion during its intestinal absorption, studied with the rat everted gut sac model. The method was validated in the concentration range of 1-15 microM (0.024-3.58 microg/mL) for bupropion and 0.005-1 microM (0.00127-0.25 microg/mL) for hydroxybupropion with 10 microL injected. Bupropion is used as a probe for the activity of the CYP2B6 isoenzyme of the P450 family of enzymes in man. Its major metabolite hydroxybupropion was found in the serosal media of the gut sac showing that the isoenzyme of the 2B group was active in the intestinal mucosa and metabolized bupropion during its passage across the mucosa. The metabolite was also quantified in the mucosal media indicating its ability to cross the apical membrane of the epithelial cells. PMID- 16246643 TI - Micellar electrokinetic capillary chromatography reveals differences in intracellular metabolism between liposomal and free doxorubicin treatment of human leukemia cells. AB - Doxil is a pegylated liposome formulation of the anthracycline doxorubicin. To better explain observed differences in the toxicity of Doxil and free doxorubicin in solution, the intracellular metabolism of the formulations after treatment in CCRF-CEM and CEM/C2 human leukemia cell lines was investigated. Using micellar electrokinetic capillary chromatography with laser-induced fluorescence detection, with a 63 zepto (10(-21)) mole doxorubicin limit of detection, five common metabolites and doxorubicin were detected upon treatment with both of these drug delivery systems. Two unique metabolites appeared with the Doxil and two unique metabolites appeared with the free doxorubicin delivery systems. For common metabolites, the relative amount of metabolite generated from Doxil was approximately 10 times higher than for free doxorubicin. PMID- 16246644 TI - Oligosaccharide analysis by capillary-scale high-pH anion-exchange chromatography with on-line ion-trap mass spectrometry. AB - A capillary-scale high-pH anion-exchange chromatography (HPAEC) system for the analysis of carbohydrates was developed, in combination with two parallel on-line detection methods of sub-picomolar sensitivity: (1) pulsed amperometric detection (PAD); (2) capillary-scale desalting followed by electrospray ion-trap (IT) mass spectrometry (MS). The capillary chromatographic system combined the superb selectivity of HPAEC that allows routine separation of isomeric oligosaccharides with the information on monosaccharide sequence and linkage positions obtained by MS/MS fragmentation using the IT-MS. The applicability of the system in biomedical research was demonstrated by its use for the analysis of a urine sample of a GM1-gangliosidosis patient. Isomeric glycans in the sample could be resolved by HPAEC and assigned on the basis of the monosaccharide linkage information revealed by on-line IT-MS/MS. PMID- 16246645 TI - A homotetrameric agglutinin with antiproliferative and mitogenic activities from haricot beans. AB - A homotetrameric agglutinin with a molecular mass of 130 kDa was isolated from seeds of the haricot bean. The agglutinin was isolated using a procedure that involved ion exchange chromatography on DEAE-cellulose, affinity chromatography on Affi-gel blue gel and gel filtration by fast protein liquid chromatography on Superdex 200. Haricot bean agglutinin was adsorbed on DEAE-cellulose and Affi-gel blue gel. The hemagglutinating activity of the agglutinin was stable up to 40 degrees C. It underwent a 40% decline when the temperature was raised to 50 degrees C and a complete loss when the temperature was further increased to 80 degrees C. The hemagglutinating activity exhibited a time-dependent loss in activity when the agglutinin was incubated at 100 degrees C for different durations. No activity was discernible when the agglutinin was left at 100 degrees C for 1 min. The activity also underwent a decline in the presence of 500 mM FeCl(3) and CaCl(2). Haricot bean agglutinin manifested a weaker mitogenic activity than concanavalin A toward mouse splenocytes. It exhibited antiproliferative activity toward the tumor cell lines M1 [leukemia], HepG2 [hepatoma] and L1210 [leukemia] cells. PMID- 16246646 TI - Some thoughts on what's ahead for GI in 2006 and beyond. PMID- 16246648 TI - Assessment of patients at intermediate cardiac risk. AB - The optimal approach to patients who are at intermediate risk for coronary artery disease is not clear. Various risk assessment scoring systems and imaging modalities can add diagnostic and prognostic value to cardiovascular evaluation. A better understanding of these available assessment tools can help physicians to optimize management and outcomes in patients who are initially categorized as at intermediate risk. Detailed case studies are presented that illustrate the use of these assessment tools in patients at intermediate risk. PMID- 16246649 TI - Functional capacity and cardiovascular assessment: submaximal exercise testing and hidden candidates for pharmacologic stress. AB - Submaximal exercise testing is often used to estimate functional capacity in non athletes, to assess cardiovascular disease in elderly or frail patients, to demonstrate exercise equipment, or to risk-stratify patients after myocardial infarction. However, submaximal exercise testing is not sufficiently sensitive, specific, or predictive to have widespread clinical utility, except in post myocardial infarction protocols. Many patients for whom submaximal exercise testing is not useful are unable to exercise sufficiently for maximal testing and are referred for imaging with pharmacologic stress. Although some patients who are unable to exercise adequately are easily recognized, many are not. The identification of such patients before they fail a maximal exercise test attempt is beneficial to both the patient and the imaging laboratory. PMID- 16246650 TI - Role of electron-beam computed tomography and nuclear stress testing in cardiovascular risk assessment. AB - Although it is well recognized that stenotic coronary artery lesions carry a significant risk for cardiac events, the importance of nonstenotic lesions is generally underappreciated. However, many acute myocardial infarctions are caused by lesions that cause <50% stenosis. Coronary artery calcification is being increasingly studied as a marker of risk for cardiac events. Measurement of coronary artery calcium using electron-beam computed tomography is emerging as a useful tool to further risk-stratify patients who are otherwise at intermediate risk for events. Coronary calcium scores have been shown to add independent predictive value to traditional risk assessment. PMID- 16246652 TI - Summary and conclusions. PMID- 16246651 TI - Sequential single-photon emission computed tomography myocardial perfusion imaging. AB - The utility of stress myocardial perfusion imaging (MPI) for the diagnosis and prognosis of coronary artery disease (CAD) has been firmly established in numerous clinical studies and has become an essential component of clinical practice. Stress MPI is now used regularly to guide initial risk stratification and management of patients with CAD. Because stress MPI provides an assessment of the physiologic significance of CAD, it is a particularly attractive procedure for assessing follow-up risk. Today, sequential stress MPI is being used increasingly to track disease progression, assess follow-up risk, detect restenosis following revascularization, and evaluate the efficacy of aggressive medical therapy and risk-factor modification. By providing serial snapshots of the disease and its impact on perfusion, sequential stress MPI may alter treatment decisions and ultimately improve long-term patient management and outcomes. Use of sequential stress MPI to detect changes in perfusion following surgical or medical therapies is being tested currently in the Clinical Outcomes Using Revascularization and Aggressive Drug Evaluation (COURAGE) and Adenosine Sestamibi Single-Photon Emission Computed Tomography Postinfarction Evaluation (INSPIRE) trials. PMID- 16246653 TI - [Evidence based surgery has some limitations]. AB - Evidence based medicine can be defined as the application of the best evidence in the care of a given patient. When applied to surgical practice, it appears that this concept has some limitations. To discuss these limitations, the authors made the choice to discuss the terms the original definition. Some factors are related to the paucity and the poor quality of randomized controlled trials and meta analyses in surgery, to the difficulties to appraise the surgical publications and apply the results of randomized trials to a given patient, and to bring the surgeons more willing to endorse the principles of evidence-based medicine. But all these limitations could be overcome making evidence-based surgery not to be a simple passing fad but a formal paradigm. PMID- 16246654 TI - Improving contraceptive technology: is it a zero-sum game? PMID- 16246655 TI - Measured and reported weight change for women using a vaginal contraceptive ring vs. a low-dose oral contraceptive. AB - BACKGROUND: Women often stop hormonal contraception because of perceived weight change. We conducted a randomized trial comparing the contraceptive vaginal ring to a low-dose oral contraceptive (OC). We examined the difference between women's reported and measured baseline weights and looked at factors affecting perceived weight change. METHODS: We randomized 201 participants to either the vaginal ring or an OC for three cycles. We weighed participants upon enrollment (n=194) and at exit (n=167), using the same instrument for all measurements. Participants also provided self-reported height and their reactions to perceived weight changes. RESULTS: Baseline weight and body mass index were similar for both groups (mean weight=145.9 lb). Measured weight was, on average, 4.4 lb more than reported weight; this difference was greater in overweight and obese participants. Participants gained an average of 2.8 lb over 3 months; this gain did not differ between groups or by baseline weight. Subjects who reported a "bad change" in weight at exit (n=34) gained an average of 4.4 lb, whereas those who reported "no change" (n=112) gained 2.2 lb and those who reported a "good change" (n=14) gained 3.3 lb. CONCLUSION: Participants underreported their weight, and this difference was greater for heavier women. There was little weight change for the women in our study. Participants' opinions about weight change were not correlated with measured weight changes. PMID- 16246656 TI - Randomized trial of mifepristone and buccal or vaginal misoprostol for abortion through 56 days of last menstrual period. AB - OBJECTIVE: Mifepristone-misoprostol medical abortion has been approved in the United States since 2000. U.S. providers have preferred to use vaginal misoprostol because of evidence that such a regimen is more effective in later gestations. Buccal administration of misoprostol may be equally effective and more acceptable to some women. METHODS: This open-label, randomized trial was conducted at two sites in Rochester, NY, and involved healthy women with pregnancies through 56 days since the last menstrual period (LMP) as indicated by sonogram. Women received mifepristone 200 mg orally and were randomized to use 800 mug of misoprostol either buccally or vaginally 1 to 2 days later. They returned within 15 days for repeat sonogram. If the woman's pregnancy had not been completely aborted by day 36, a suction abortion was performed. The primary outcome was a complete abortion without surgical intervention. RESULTS: Four hundred forty-two women were enrolled in the study, and complete data were available on 429. The efficacy rate was 95% (205/216) in the buccal group and 93% (199/213) in the vaginal group (chi(2)=0.43, p=.51). Nausea was the most commonly reported side effect, affecting 70% in the buccal group and 62% in the vaginal group. There were no differences in the satisfaction with the overall procedure between the buccal (92%) and the vaginal groups (95%) (chi(2)=1.87, p=.17). CONCLUSION: Buccal administration of misoprostol after low-dose mifepristone for medical abortion appears to be a highly effective and acceptable alternative compared with vaginal administration for medical abortion in pregnancies through 56 days LMP. PMID- 16246657 TI - Same-day initiation of the transdermal hormonal delivery system (contraceptive patch) versus traditional initiation methods. AB - INTRODUCTION: Published comparisons of oral contraceptive pill (OCP) initiation methods demonstrate that OCP initiation at the office visit ("Quick Start") resulted in higher continuation rates into the second cycle. This trial was performed to investigate whether Quick Start with the contraceptive patch would provide similar results. MATERIALS AND METHODS: Sixty women were randomized to initiate use of the contraceptive patch using Quick Start (Group 1, n=30) or on the first day of their next menses (Group 2, n=30). Telephone contact at 6 weeks occurred to ensure that the second cycle had been initiated. A single follow-up visit was scheduled after completion of the third patch cycle. RESULTS: Continuation rates for Groups 1 and 2 were 97% and 93% (p=1.0), respectively, into the second cycle, and 93% and 90%, respectively, into the third cycle (p=1.0). Only approximately half of the subjects planned to continue using the patch after the study. CONCLUSION: Quick Start for the contraceptive patch did not improve continuation rates into the second or third cycle. PMID- 16246658 TI - TCu 380A IUD: a reversible permanent contraceptive method in women over 35 years of age. AB - OBJECTIVE: The objective of the study was to evaluate the performance of the TCu 380A IUD in women who had been using the device for more than 10 years and who were 35 years of age or more on completion of the 10th year of IUD use. METHODS: A total of 228 women who had an IUD inserted between 1987 and 1992 were included in the study. The cutoff date for analysis was January 31, 2004. Clinical performance was evaluated by life-table analysis. The mean age of women at 10 years of use was 38.8+/-0.4 years and mean parity was 2.2+/-0.08 (mean+/-SD). The duration of follow-up beyond 10 years ranged from 1 to 72 months. No pregnancy was observed in 366 woman-years of observation beyond 10 years of use. The main reason for discontinuation was removal of the device because the clients had previously been informed that the IUD was not approved for use beyond 10 years. This reason accounted for a gross cumulative 6 years discontinuation rate of 42.5 per 100 women beyond 10 years. The other main reasons for discontinuation beyond 10 years of use were surgical sterilization, menopause and expulsion with gross cumulative 6-year termination rates of 19.2, 11.0 and 21.2 per 100 women, respectively. The cumulative continuation rate beyond 10 years was 67.0 at the end of the first year of follow-up and 21.2 at the end of the sixth year. CONCLUSION: We found no evidence that the TCu 380A IUD loses its effectiveness after 10 years of use. RESULTS: The concept that women who have insertion of a TCu 380A IUD at the age of 25 years or older could use this IUD as a reversible but permanent method of contraception up to the menopause continues to be supported by the accumulation of evidence, although definitive evidence remains to be obtained. PMID- 16246659 TI - Five years' experience with a small intracervical/intrauterine levonorgestrel releasing device. AB - OBJECTIVE: A randomized study was performed to compare the efficacy, safety and acceptability of a new model of an intracervical/intrauterine contraceptive device (ICD) releasing 20 microg of levonorgestrel (LNG) per day. METHODS: The LNG-ICD was inserted in Group I into the cervical canal and in Group II into the uterine cavity. Group I included 151 women (age, 18-43 years) whereas Group II included 147 (age, 19-43 years). The number of nulliparous women was 145. RESULTS: The 5-year results are presented here. The results showed a total continuation rate of 50%; the continuation rate in the cervical group and that in the uterine group were 53.6% and 46.3%, respectively--the difference being statistically insignificant (p=.3593). The main reason for termination was a wish for pregnancy, which is explained by the relatively young age and degree of nulliparity of the study population. During the first year, two pregnancies occurred in both groups. Two of these were ectopic, one in each group. The other two occurred after unnoticed expulsions. Thereafter, no pregnancies occurred. The cumulative gross rate for pregnancy was 1.3 and the Pearl index at 5 years was 0.425. The total expulsion rate was relatively high (11.1%). Expulsions occurring during the first few months of the first year were related to insertion. Removals because of bleeding and because of amenorrhea were low, the combined gross rate being 5.7 and the Pearl rate 1.8 at 5 years. Also, the gross rate of infection was low (0.7). The continuation was high in spite of a high rate of removals for planning pregnancy (15.4). CONCLUSIONS: The method is safe and effective. There were only minor differences between the groups. There were no perforations and the incidence of infection was low. The device can also be used by young nulliparous women. PMID- 16246660 TI - A comparative study of the levonorgestrel-releasing intrauterine system Mirena versus the Copper T380A intrauterine device during lactation: breast-feeding performance, infant growth and infant development. AB - BACKGROUND: Mirena is a levonorgestrel-releasing intrauterine system (LNG-IUS) that provides highly effective and long-acting progestogen-only contraception. OBJECTIVE: The objective of this study was to analyze the possible effects of using LNG-20 microg IUS on breast-feeding performance, infant growth and infant development during the first postpartum year as compared with the Copper T380A intrauterine device (Cu T380A IUD). DESIGN: This study is a prospective, controlled and randomized trial. SETTING: The study was conducted at the Department of Obstetrics and Gynecology, Assiut University Hospital, Egypt. METHODS: Three hundred twenty lactating women asking for initiation of contraception during the early postpartum stage were assigned randomly into two groups, the LNG-20 microg IUS group (n=163) and the Cu T380A group (n=157). The insertions were done 6-8 weeks postpartum. Each participant was followed up at three monthly intervals after insertion and until the first birthday of her baby. During these visits, the breast-feeding pattern was assessed, certain infant physical growth parameters were measured and a set of infant development tests was performed. RESULTS: No pregnancy occurred in both groups. There were no significant differences in the net continuation rates between the two groups (89.3 for LNG-IUS vs. 90.9 for Cu T380A). The LNG-20 microg IUS group had comparable rates of breast-feeding continuation, complete weaning, full breast feeding and partial breast-feeding, with the Cu-IUD group. No statistically significant differences were found between groups with regard to all infant physical growth parameters and various infant development tests. CONCLUSION: The findings of the current study confirm that the use of LNG-20 microg IUS during the first postpartum year in lactating women provides highly effective and acceptable contraception and does not negatively influence breast-feeding or the growth and development of breast-fed infants. PMID- 16246661 TI - Extended use of a progesterone-releasing vaginal ring in nursing women: a phase II clinical trial. AB - AIM: This study evaluates the performance of extended use of a progesterone (P) releasing vaginal ring (PVR) in nursing women. METHOD: An open-label, noncomparative study on the safety and contraceptive efficacy of PVR replaced every 4 months of use (instead of 3 months) in 192 PVR acceptors. PVR use was initiated at day 59+/-2 (mean+/-SD) postpartum and continued until weaning or completing the use of three PVRs. RESULTS: Plasma P levels attained with the ring decreased from 17+/-1 to 14+/-1 nmol/L (mean+/-SE) from the third to the fourth month of use. These levels are still over the critical level of 10 nmol/L required for contraceptive protection. One pregnancy occurred in the third month of use of the second ring in 1998 woman-months of exposure. Extended use of the ring did not appear to affect breast-feeding performance or the rate of infant growth, and lactational amenorrhea was prolonged. No differences in the characteristics of bleeding between the third and fourth month of ring use were observed. CONCLUSION: The results indicate that use of the PVR for 4 months represents a safe and effective contraceptive for nursing women. PMID- 16246662 TI - A randomized clinical trial of the addition of laminaria to misoprostol and hypertonic saline for second-trimester induction abortion. AB - OBJECTIVE: To compare the outcomes of second-trimester induction abortion with misoprostol and hypertonic saline, with and without use of laminaria. METHOD: Fifty-eight women, between 17.5 and 22.5 weeks' gestation, were randomly assigned to receive or omit laminaria in conjunction with other procedures for induction abortion. All women received a fetocidal dose of 60 cc intra-amniotic hypertonic saline. If the woman was to receive laminaria, they were inserted next. This was followed by vaginal misoprostol 200 mug, which was repeated every 6 h. RESULT: Women with laminaria inserted before misoprostol administration had longer intervals from start of misoprostol to delivery of the fetus (induction times) than women without laminaria. Induction time was 14.4 vs. 11.4 h, respectively (p=.04, Wilcoxon rank sum test). Total misoprostol use was higher in the laminaria group, 628 mug (95% CI, 516-738) vs. 496 mug (95% CI, 419-573) (p=.05). Total analgesic use was also higher in the laminaria group, 41 mg of morphine (95% CI, 32-50) vs. 26 mg of morphine (95% CI, 18-32) (p=.02). CONCLUSION: Laminaria use, in conjunction with misoprostol and hypertonic saline, significantly prolongs induction time and increases narcotic analgesia usage. PMID- 16246663 TI - The combined contraceptive vaginal ring (NuvaRing) and endometrial histology. AB - We investigated the effects of NuvaRing on endometrial histology in a 2-year open label, multicenter trial in 103 premenopausal women aged 18-35 years. Subjects received 26 cycles of treatment, each comprising 3 weeks of ring use followed by a 1-week ring-free period. Endometrial biopsies were taken at baseline, and at cycles 13 (month 12) and 26 (month 24). Normal biopsy results were obtained in all subjects after 24 months of NuvaRing treatment. Atrophic or inactive endometrium and secretory changes were present in the majority of the biopsies after 1 year and 2 years of treatment with the vaginal ring. It was concluded that long-term use of NuvaRing has no adverse effects on endometrial histology. PMID- 16246664 TI - Comparative acceptability study of the Reality female condom and the version 4 of modified Reddy female condom in India. AB - OBJECTIVE: Affordable, acceptable and effective female controlled options are required worldwide for prevention of human immunodeficiency virus (HIV) infection and other sexually transmitted diseases. We carried out a comparative acceptability study of Reality and Reddy (version 4) female condoms. METHODS: Sixty eligible couples were enrolled and randomly assigned to use either Reality or Reddy condom first. They used three Reality condoms and three Reddy condoms each with at least one condom use per week. RESULTS: Reddy female condom had a significantly better acceptability than Reality condom among women who were less educated and who had not used male condom before. In spite of higher acceptability score, participants were less confident about the Reddy condom for protecting them from HIV disease or pregnancy as compared to a male condom. CONCLUSIONS: Female condoms are being introduced in India. This study has generated data that is suggestive of optimism for this female controlled option. PMID- 16246665 TI - Consultations concerning contraception and induced abortions among university students--trends in Finland 1986-2003. AB - The Finnish Student Health Service (FSHS) provides primary health care services to university students in Finland. The material for this study was drawn from the statistics of the FSHS. From 1986 to 2003, the number of students increased by 50%, while physician consultations did not, indeed to the contrary. The numbers of family planning consultations fell from 358 to 217 per 1000 female students. The induced abortion rates among university students are very low compared with the population in general and decreased from 4 to 3 per 1000 female students. Students are responsible, and because they want to postpone pregnancy after studies, they use contraceptives effectively. The FSHS has succeeded in guaranteeing contraception services and in preventing unwanted pregnancies and minimizing the need for induced abortions among university students while at the same time decreasing physician consultations. This was possible by changing the distribution of tasks among physicians and nurses, and by adding telephone contacts instead of face-to-face consultations with physicians. PMID- 16246666 TI - Scaling up postabortion contraceptive service--results from a study conducted among women having unwanted pregnancies in urban and rural Tanzania. AB - INTRODUCTION: It is well recognized that unwanted pregnancies and unsafe abortion are significant public health problems in sub-Saharan Africa. At the International Conference on Population and Development held in Cairo in 1994, postabortion care was prioritized as a means to reduce maternal morbidity and mortality associated with unsafe abortion. However, only a few postabortion care programs have been implemented and most of them have been confined to urban settings. The present study describes the magnitude of the problem of unwanted pregnancies among women with incomplete abortion in urban and rural Tanzania and evaluates the outcome of a postabortion care intervention. METHODS: Data were collected among 781 women admitted with incomplete abortion in Dar es Salaam region (urban Tanzania) and 575 women in Kagera region (rural Tanzania). RESULTS: Sixty-seven percent of the women in urban Tanzania and 42% in rural Tanzania stated that their pregnancy was unwanted. Contraceptive acceptance among women with unwanted pregnancies was high; 93% in urban Tanzania and 71% in rural Tanzania left with a contraceptive method. CONCLUSION: The high proportion of women with unwanted pregnancies in urban and rural Tanzania underlines the need of scaling up postabortion contraceptive service. PMID- 16246667 TI - Steroid receptor expression in vaginal epithelium of healthy fertile women and influences of hormonal contraceptive usage. AB - BACKGROUND: The objective of this study was to evaluate whether long-term usage of hormonal contraceptives modifies the steroid receptor expression in the human vaginal epithelium of healthy young women. METHODS: In a cross-sectional study, three groups of hormonal contraceptive users [combined oral contraceptives (COCs), levonorgestrel implants (LNG) and depot medroxyprogesterone acetate injections (DMPAs)] were compared to controls. Fifteen subjects (20-34 years) were enrolled to each group. Vaginal biopsies were collected at two occasions from each subject, and serum concentrations of E(2) and progesterone were measured. Monoclonal antibodies directed against progesterone receptors (PRs) and estrogen receptors (ERs) were used in immunohistochemistry on formalin-fixed tissue sections of vaginal mucosa. A program for immunohistomorphometric quantification was devised to estimate frequency of epithelial steroid receptor expressing cells. RESULTS: Progesterone receptor expression was markedly down regulated and significantly reduced in DMPA users compared to controls, COC and LNG users. In DMPA users, the ER expression was significantly elevated in the first compared to the second sample, and significantly elevated compared to LNG users. Estradiol concentration in serum was significantly reduced in hormonal contraceptive users compared to controls. CONCLUSIONS: Steroid receptor expression in human vaginal epithelium is altered by long-term use of DMPA compared to controls. PMID- 16246668 TI - Risks of mifepristone abortion in context. PMID- 16246670 TI - Description of emergency contraception in the media. PMID- 16246671 TI - Emergency contraception--a different interpretation. PMID- 16246674 TI - ASGE Guideline: the role of endoscopy in the patient with lower-GI bleeding. PMID- 16246673 TI - ASGE Guideline: the role of endoscopy in the management of variceal hemorrhage, updated July 2005. PMID- 16246675 TI - Transnasal endoscopy vs. fluoroscopy for the placement of nasoenteric feeding tubes in critically ill patients. AB - BACKGROUND: Placement of a nasoenteric feeding tube (NET) beyond the pylorus in critically ill patients is desirable. Bedside methods are unreliable, and fluoroscopic methods require transport and/or radiation exposure. Traditional endoscopic methods require sedation and oronasal transfer techniques. Transnasal techniques of NET placement by using recently developed ultrathin endoscopes have been described. The object of this prospective study was to compare the efficacy of NET placement by using ultrathin transnasal endoscopy vs. fluoroscopic placement. METHODS: This is a prospective randomized study of endoscopic vs. fluoroscopic NET placement. The settings of the study were intensive care units at academic medical center. The study included 100 consecutive patients who required NET placement. They received endoscopic NET placement at the bedside with a 5.1-mm, ultrathin endoscope by using a transnasal over-the-wire technique vs. fluoroscopic NET placement by using standard techniques. The procedure success was defined as postpyloric (beyond the duodenal bulb) NET placement, jejunal placement success, and procedure time. RESULTS: Tube placement success was not significantly different between endoscopic and fluoroscopic methods (90% with both methods; p = 1.00). The endoscopic procedure duration (12.8 +/- 6.4 minutes) was significantly shorter than fluoroscopic procedure duration (19.3 +/- 12.0 minutes) (p < 0.001). Procedure duration decreased significantly (from 17.3 +/- 6.2 minutes to 8.0 minutes +/- 4.2 minutes, p = 0.04), and jejunal placement increased significantly (from 60% to 100%, p = 0.04) from the first to the last 10 endoscopic procedures. CONCLUSIONS: NET placement success with an ultrathin transnasal endoscope is equivalent to fluoroscopic placement with faster procedure times. More distal placement and procedure times improve with increasing experience with the endoscopic technique. Endoscopic NET placement can be performed at the bedside without the need for oronasal transfer, additional sedation, or fluoroscopy. PMID- 16246676 TI - Another indication for transnasal, unsedated upper-GI endoscopy. PMID- 16246677 TI - Early institution of pre-cutting for difficult biliary cannulation: a prospective study comparing conventional vs. a modified technique. AB - BACKGROUND: Pre-cutting techniques have been used to gain biliary access at the expense of an increased complication rate. This may be because of the multiple attempts to achieve cannulation by using standard methods before pre-cutting and causing excess edema and papillary trauma. There are limited data on the early use of pre-cutting techniques. METHODS: We performed a prospective study of the early introduction of needle-knife techniques in patients with difficult biliary cannulation. Standard biliary cannulation was attempted with a sphincterotome and a guidewire. If this failed within 10 minutes or if there were more than 5 pancreatic cannulations, the needle-knife technique was used. Either a standard method of pre-cutting (below-upward) from the papillary orifice or the modified technique of pre-cutting (above-downward), stopping short of the papillary orifice, was adopted, as per the discretion of the endoscopist. If pre-cutting failed, the cannulation was reattempted 24 to 48 hours later. RESULTS: A total of 346 therapeutic biliary ERCP procedures were performed between April and August 2003. Of these, 70 patients (20%) (mean age, 54 years; 38 men) underwent needle knife pre-cut sphincterotomy (16 with the standard technique). In 58 patients (83%), the procedure was successful with the initial pre-cutting, making the total success at initial ERCP 334/346 (96.5%). Nine patients in whom pre-cut failed, returned for a second-attempt ERCP, with 7 completed successfully. The total success rate of pre-cutting was 65/70 (93%). The overall success rate of biliary cannulation, after two ERCP attempts, was 341/346 (98.5%). Six patients had mild bleeding, and one had mild pancreatitis. There was no difference in these complications between the two types of pre-cut techniques. CONCLUSIONS: The early use of needle knife for difficult biliary cannulation is safe and effective, irrespective of the technique used. PMID- 16246678 TI - Clinical predictors of Barrett's esophagus length. AB - BACKGROUND: Assessment of clinical factors associated with Barrett's esophagus (BE) length remained within the realm of anecdotal reports or one center's experience. The aim of this multicenter study was to determine which clinical factors are highly correlated with the length of BE. METHODS: Patients diagnosed with BE were recruited into the study from 5 academic centers in the United States. All patients had an upper endoscopy that documented BE by the presence of intestinal metaplasia in biopsy specimens. All patients were evaluated by a validated demographic questionnaire and the GERD Symptom Checklist. RESULTS: A total of 263 patients with BE were recruited into the study. Mean BE length length for the entire sample was 4 +/- 3.3 cm. A longer hiatal hernia (r = 0.22, p < 0.01), any dysplasia (t = -2.3, p < 0.05), H2 receptor antagonist (H2-RA) consumption (t = 1.98, p < 0.05), and nonsmoking (t = -2.5, p < 0.05) were correlated with a longer segment of BE. Proton pump inhibitors (PPI) (t = 1.96, p < 0.05) were correlated with a shorter segment of BE. CONCLUSIONS: PPIs were correlated with shorter lengths of BE. In contrast, a longer hiatal hernia, any dysplasia, nonsmoking, or use of H2-RAs were correlated with a longer BE segment. PMID- 16246679 TI - Should we worry about the length of Barrett's esophagus? PMID- 16246680 TI - A fluorescence confocal endomicroscope for in vivo microscopy of the upper- and the lower-GI tract. AB - BACKGROUND: This report describes the development and the clinical evaluation of a novel confocal endomicroscope for obtaining fluorescence images of cellular morphology of the mucosae of the upper- and the lower-GI tract in vivo. The work assessed the feasibility of performing in vivo microscopy at endoscopic examination and evaluated fluorescence imaging protocols. METHODS: Images were collected in real time by using two prototype endoscope configurations, featuring slightly different miniaturized fiber-optic confocal microscopes, fitted integrally into the tips of conventional endoscopes. Confocal scanning was performed at 488 nm illumination for excitation of exogenously applied fluorophores (topical acriflavine and intravenous fluorescein). The images were compared with conventional histology of biopsy specimens and the findings of white-light endoscopy. RESULTS: Confocal endomicroscopy enabled imaging of cellular and subcellular structures (i.e., nuclei) of the GI tract. The crypts of the colonic mucosa, the villi of the terminal ileum and duodenum, the gastric pits of the stomach, and the squamous epithelium of the distal esophagus could be clearly visualized. Acriflavine strongly contrasted the cell nuclei of the surface epithelium, including the absorptive epithelial cells and the mucous secreting goblet cells. Fluorescein stained the extracellular matrix of the surface epithelium and also the subepithelial layers of the lamina propria. Images at increasing depth beneath the epithelium showed the mucosal capillary network. The findings correlated with the histology of biopsy specimens. CONCLUSIONS: The development of a fluorescence confocal endomicroscope makes it practical to examine the upper- and the lower-GI mucosa in cellular detail during otherwise routine endoscopic examination. The results represent a major technical advance in the development of this new optical imaging modality for the in vivo examination of GI tissue. PMID- 16246681 TI - Confocal microscopy from the bench to the bedside. PMID- 16246682 TI - Lugol chromoendoscopy as a diagnostic tool in so-called endoscopy-negative GERD. AB - BACKGROUND: Esophageal mucosal breaks are found in less than half of patients with typical reflux symptom. Thus, endoscopy appears to be an insensitive test for GERD. Lugol chromoendoscopy has been used to detect early esophageal cancer, which is difficult to recognize by routine observation without dye staining. The aim of this study was to determine the efficacy of Lugol chromoendoscopy in the diagnosis of so-called endoscopy-negative GERD (ENRD). METHODS: The study was conducted with 61 patients (21 women; mean age of 59.8 years) with reflux symptoms and 42 controls (15 women; mean age, 65.0 years). In the absence of any esophageal mucosal abnormalities at conventional endoscopy, Lugol's iodine solution was sprayed onto the esophageal surface, followed by evaluation of the staining pattern. When Lugol-unstained streaks were observed at chromoendoscopy, biopsy specimens were obtained from unstained streaks and from adjacent stained mucosa. Histologic evaluation included basal cell hyperplasia, papillary length, and cellular infiltration. RESULTS: Twenty-two (36%) of 61 patients with reflux, and 4 (10%) of 42 controls had visible esophagitis by conventional endoscopy. Lugol chromoendoscopy was performed in the remaining 39 patients and 38 controls. The entire esophageal mucosa was uniformly stained dark brown in 20 patients with reflux and 37 controls. In the remaining 19 patients with reflux and in one control, several unstained streaks were observed in the distal esophagus (p < 0.0001). Histologically, Lugol-unstained mucosa showed a significantly thicker basal cell layer (30.9% +/- 7.6% vs. 12.3% +/- 4.5% of total epithelial thickness, mean +/- standard deviation [SD], p < 0.01) and longer papillae (57.9% +/- 12.6% vs. 38.1% +/- 12.6% of total epithelial thickness, mean [SD], p < 0.01) compared with stained mucosa. In addition, infiltration of lymphocytes in the epithelium was significantly increased in unstained mucosa than in stained mucosa (p < 0.01). CONCLUSIONS: Visible unstained streaks by Lugol chromoendoscopy seem to be indicative of mucosal injury, which was not detectable by conventional endoscopy. Lugol chromoendoscopy is simple and could be useful for the diagnosis of ENRD. This method could be appealing for the endoscopist as it is easy, safe, and can be performed at the same endoscopic session. PMID- 16246683 TI - The color of reflux: confessions of a skeptic. PMID- 16246684 TI - Incidence of abdominal wall metastasis complicating PEG tube placement in untreated head and neck cancer. AB - BACKGROUND: Abdominal wall metastasis after PEG tube placement has been reported in patients with head and neck cancer. The incidence of this occurrence is unknown. OBJECTIVE: Evaluation of the incidence of abdominal wall metastasis as a complication of PEG tube placement in patients with head and neck cancer. DESIGN: Retrospective chart review. SETTING: H. Lee Moffitt Cancer Center and Research Institute, Nutritional Support Services. SUBJECTS: Head and neck cancer patients requiring nutritional support with PEG tube placement. RESULTS: Of the 304 patients with head and neck cancer, 218 had active disease with a viable tumor in the oropharynx or hypopharynx at the time of PEG placement. Two of these patients, both with active disease (0.92%), developed a PEG site metastasis. CONCLUSION: There is a small but definite risk for tumor implantation in the gastrostomy site when using the pull technique in patients with active head and neck cancer. Careful assessment of the oropharynx and hypopharynx before PEG tube placement and the use of alternative techniques for enteral access in patients with untreated or residual malignancy are recommended to minimize this risk. Use of other percutaneous techniques that do not involve traversing the hypopharynx with the catheter may help to prevent tumor translocation. When head and neck cancers metastasize to the gastrostomy site, patient survival appears limited even with extensive resection. PMID- 16246685 TI - Complications, limitations, and failures of capsule endoscopy: a review of 733 cases. AB - BACKGROUND: Although a variety of technical and clinical problems of capsule endoscopy have been reported, their incidence and clinical importance are unknown. The objective was to evaluate the incidence and the type of such events. METHODS: This was a retrospective analysis of 733 consecutive examinations at 4 large referral centers. The main outcome measurements were that the problems were classified as "technical," i.e., related to the functioning of the equipment, and "clinical," i.e., related to patient characteristics. For each type of event, the causes and the impact on the ability to reach a diagnosis were examined. RESULTS: A total of 183 problems were recorded in 174 patients (23.7%): one problem occurred in 165 patients, two in 9 patients. In 8.46% of patients, technical limitations or failures, or clinical complications prevented or hampered diagnosis. Technical problems (e.g., gaps in the recordings, short duration of capsule batteries, failure of downloading) occurred in the early phase of capsule use in 8.6% of examinations and prevented or hampered diagnosis in 2.9%. Clinical problems (difficulty/inability to swallow the capsule, incomplete small-bowel examination) occurred in 16.4% of examinations and hampered or prevented diagnosis in 6.4%. Capsule retention that required surgical or endoscopic retrieval occurred in 1.9% of cases. CONCLUSIONS: Technical problems were rare and hampered or prevented the diagnosis in a very small number of cases. The majority of clinical failures were related to an incomplete small-bowel examination. PMID- 16246686 TI - Impact of capsule endoscopy in obscure small-bowel bleeding: defining strict diagnostic criteria for a favorable outcome. AB - BACKGROUND: The most frequent indication for capsule endoscopy is to diagnose the cause of obscure GI bleeding. The objective of the study was to determine the impact of capsule endoscopy on the outcome of patients with GI bleeding of obscure origin. METHODS: Ninety-six patients (53 men, 43 women; mean [standard deviation] age, 60.84 years [16.55 years]) were enrolled in the study. All patients have been subjected to gastroscopy, colonoscopy, small-bowel barium follow-through or enteroclysis, and push enteroscopy; no bleeding site had been identified. Capsule endoscopy was performed with the Given M2A video capsule system. By using strict criteria, studies were classified as having positive findings, findings of uncertain significance, and no findings. Outcome was defined as continued or complete resolution of bleeding. RESULTS: Positive findings, findings of uncertain significance, and no findings were identified in 41.7%, 20.8%, and 37.5% of our study population, respectively. The most common lesions seen were angiodysplasias of the small intestine. Therapeutic intervention was possible in 82.5% of patients with positive findings and in 35.0% of patients with findings of uncertain significance. Complete resolution of bleeding, after a median (interquartile range) follow-up period of 14 months (9 17 months), occurred significantly more often in patients with positive findings (68.4%) compared with patients with findings of uncertain significance and no findings (40.8%, p = 0.009). CONCLUSIONS: Capsule endoscopy increases the diagnostic yield in the workup of patients with obscure small-bowel bleeding. Strictly defined positive findings are associated with a favorable outcome. PMID- 16246688 TI - Yield of EUS-guided FNA of pancreatic masses in the presence or the absence of chronic pancreatitis. AB - BACKGROUND: Evaluation of a focal pancreatic mass in the setting of chronic pancreatitis (CP) is a diagnostic challenge. The objectives of the study were to compare the diagnostic yield and accuracy of EUS-guided FNA (EUS-FNA) in the evaluation of pancreatic-mass lesions in the presence or the absence of CP and to identify predictors of CP before EUS-FNA of pancreatic-mass lesions. METHODS: The study design was analysis of data collected prospectively on all patients with solid pancreatic-mass lesions who underwent EUS-FNA at a tertiary referral center. A total of 282 consecutive patients underwent 300 EUS-FNA procedures of pancreatic-mass lesions over a 3-year period. The diagnostic yield and the accuracy of EUS-FNA was compared between patients with and without CP. CP was defined by the presence of more than 4 EUS criteria. RESULTS: Final diagnosis was adenocarcinoma in 210 (70%), benign disease in 64 (21%), other pathology in 19 (6%), and indeterminate in 4 (2%); 3 patients (1%) were lost to follow-up. CP was noted in 75/300 (25%) patients. A lower sensitivity for EUS-FNA was observed in patients with CP than in those without CP (73.9% vs. 91.3%; p = 0.02). While patients with CP had a higher negative predictive value (88.9% vs. 45.5%; p < 0.001), no significant differences were observed for specificity (100% vs. 93.8%), positive predictive value (100% vs. 99.5%), and accuracy (91.5% vs. 91.4%) between those with and without CP. False-negative cytology was encountered in 24 cases: 6/71 (8%) with CP vs. 18/222 (8%) without CP. Patients with CP required more EUS-FNA passes to establish a diagnosis vs. those without CP (median, 5 vs. 2; p < 0.001). On multivariable analysis, age < 50 years (p < 0.001), male gender (p < 0.001), black race (p = 0.001), and the absence of jaundice (p = 0.005) were significantly associated with CP. The impact of EUS-FNA on long-term clinical management was not analyzed. The impact of individual EUS features of CP on sensitivity of EUS-FNA was not evaluated. By protocol, mass lesions that were benign required more passes to definitively exclude malignancy. CONCLUSIONS: EUS-FNA has a low sensitivity for pancreatic-mass lesions in the setting of CP. This decreased sensitivity can be overcome by performing more numbers of passes at FNA, which improves diagnostic accuracy. Demographic features and clinical presentation are predictive of underlying CP in patients with pancreatic-mass lesions. PMID- 16246687 TI - EUS-guided ethanol injection of normal porcine pancreas: a pilot study. AB - BACKGROUND: Ethanol injection is a potential means of EUS-guided pancreatic tissue ablation. The effects of injected ethanol on normal pancreas are unknown. METHODS: Transgastric ethanol injection of the body and the tail of normal porcine pancreas was performed under EUS guidance in 8 anesthetized Yorkshire pigs: 98% ethanol was injected in 4 animals, and 50% ethanol was injected in 4 animals. Serum amylase levels were monitored, and EUS was repeated on day 7. Gross and histologic examination of the pancreas was performed on day 7. RESULTS: All 8 animals tolerated ethanol injection without signs of distress. An increase in amylase levels was seen in all animals. Focal areas, 2 to 6 mm, of inflammation, necrosis, and fibrosis were observed after 50% ethanol injection. Larger (8-30 mm) areas of inflammation, necrosis, and fibrosis were seen after 98% ethanol injection, as well as one fluid collection and one colonic stricture. CONCLUSIONS: Injection of 50% ethanol into normal porcine pancreas results in focal inflammation, necrosis, and fibrosis at the injection site. Ethanol injection (98%) produces larger areas of inflammation, fibrosis, and necrosis, with local complications of pancreatitis. PMID- 16246689 TI - Diagnosing pancreatic malignancy in the setting of chronic pancreatitis: is there room for improvement? PMID- 16246690 TI - Efficacy and safety of the covered Wallstent in patients with distal malignant biliary obstruction. AB - BACKGROUND: The aim of this study was to evaluate the efficacy and the safety of the covered Wallstent, a commercially available silicone-covered, self-expandable metallic stent (covered Wallstent), for the treatment of distal malignant biliary obstruction. METHODS: Sixty-nine consecutive patients with unresectable distal malignant biliary obstruction who underwent covered Wallstent placement between October 2001 and October 2003 were studied at 4 affiliated hospitals. RESULTS: Mean stent patency and mean survival were 139.1 days and 200.9 days, respectively. Stent occlusion occurred in 7 cases (10.1%), but tumor ingrowth was not observed. Seventeen complications, including 5 cholangitis (7.2%), 4 stent migration (5.8%), 4 cholecystitis (5.8%), and 4 pancreatitis (5.8%) were observed in 16 patients (23.2%). Stent misplacement was observed in 7 patients (10.1%). CONCLUSIONS: Covered Wallstent is effective in preventing tumor ingrowth for the treatment of distal malignant biliary obstruction. However, the overall complication rate may be higher than uncovered self-expanding metal stents and covered self-expanding metal stents of different design. PMID- 16246691 TI - Medical malpractice. PMID- 16246692 TI - Solitary rectal ulcer syndrome: endoscopic spectrum and review of the literature. PMID- 16246693 TI - Improved preoperative tumor staging by 5-aminolevulinic acid induced fluorescence laparoscopy. AB - BACKGROUND: We report our data in 35 patients who underwent preoperative conventional and fluorescence-based staging laparoscopy. We use the data to address the questions of whether fluorescence examination increases the yield of metastatic lesions and alters treatment intervention. METHODS: Fluorescence laparoscopy was successfully performed in 30 patients with GI malignancies. After sensitization with 5-aminolevulinic acid, conventional white-light mode and fluorescence-light laparoscopies were sequentially performed. A suspected malignancy was biopsied. OBSERVATIONS: In 5 patients, examinations were incomplete because of adhesions. In 9 of 10 patients, hepatic or peritoneal metastases were detected by white-light examination. In 4 of these 9, blue-light examination yielded more metastatic lesions. In one patient with no lesions by white- or blue-light examination, surgery revealed hepatic metastasis in a location not accessible to laparoscopic examination. In 18 patients, surgery confirmed the absence of metastatic lesions. CONCLUSIONS: A fluorescence, blue light examination yielded more lesions than the conventional white-light examination but did not alter treatment intervention and did not enhance yield when metastatic lesion is in an inaccessible location. Continued research should focus on whether treatment intervention will be altered by the fluorescence examination. PMID- 16246694 TI - EUS-guided fine needle tissue acquisition by using high negative pressure suction for the evaluation of solid masses: a pilot study. AB - BACKGROUND: The capability of obtaining tissue samples for histologic examination during EUS has theoretical advantages over cytology alone. The objective was to evaluate the feasibility and the yield of EUS-guided FNA tissue acquisition (EUS FNTA) by using high negative pressure suction. METHODS: The study design is a prospective, observational pilot study set at a tertiary referral center. Twenty seven patients with a solid mass amenable to sampling with EUS were included in the study. FNA with a 22-gauge needle was used for a total of 5 passes. An additional pass with the same needle was performed by applying continuous high negative pressure suction using the Alliance II inflation system. The main outcome measurements were the rate of tissue acquisition and the diagnostic accuracy of EUS-FNTA. OBSERVATIONS: Tissue samples were obtained in 26 of the 27 patients (96%). Malignancy was detected in 20 of the 26 biopsy specimens obtained by FNTA and in 20 of the 27 FNA specimens. In 3 patients, EUS-FNTA failed to disclose malignancy, which in two of the patients was diagnosed by FNA. Conversely, EUS-FNTA diagnosed a recurrent malignant thymoma and a schwannoma in two FNA-negative patients. In 3 patients with both FNTA and FNA negative for malignancy, a definitive diagnosis could not be established. Overall, diagnostic accuracy was 76.9% for both EUS-FNTA and EUS-FNA. When combined, a correct diagnosis was achieved in 84.6% of the patients. Immunostaining of the retrieved tissue allowed characterization of the primary tumor in 5 cases and the diagnosis of a schwannoma and two neuroendocrine tumors. Limitations of the study were small sample size and a pilot study. CONCLUSIONS: EUS-FNTA has a high yield for the retrieval of core tissue samples. Further studies in which EUS-FNTA is performed before FNA and with variable number of passes are needed to better define its diagnostic role and performance characteristics. PMID- 16246695 TI - EUS-guided interstitial brachytherapy of the pancreas: a feasibility study. AB - BACKGROUND: Intraoperative interstitial brachytherapy has been effective when used at laparotomy to improve local control in locally advanced pancreatic cancer. Our aim in this study was to investigate the feasibility and the safety of EUS-guided brachytherapy of the pancreas in a porcine model. METHODS: A modified 18-gauge needle with radioactive seeds was inserted, under EUS guidance, into the pancreas. The radioactive seeds were implanted into the tissue by the needle. After 14 days of clinical observation, the animals were euthanized, and the tissue response to brachytherapy was examined. OBSERVATIONS: All the seeds were successfully implanted, and no migration occurred. Localized tissue necrosis and fibrosis was achieved in the pancreas, without significant complication. One pig had mild hyperlipasemia. Biochemical parameters were normal in the remaining pigs. CONCLUSIONS: EUS-guided implantation of radioactive seeds is a safe, simple, and minimally invasive technique for interstitial brachytherapy. PMID- 16246696 TI - Primary jejunal cancer. PMID- 16246697 TI - Duodenal perforation after endoscopic hemoclip application for bleeding from Dieulafoy's lesion in a duodenal diverticulum. PMID- 16246698 TI - Hookworm detected by capsule endoscopy. PMID- 16246699 TI - Intramural colonic hematoma: complication of anticoagulation with heparin. PMID- 16246700 TI - Factors that affect gastric passage of video capsule. AB - BACKGROUND: Gastric retention and delayed gastric emptying of the video capsule are major limitations. We retrospectively studied gastric transit time, gastric retention, and completeness of capsule endoscopy (CE) in relation to the conditions in which it was performed. METHODS: From May 2002 to September 2004, CE was performed in 190 patients (92 men; mean age, 58.4 years, range 16-91 years). Patients were stratified according to the way CE was performed (hospital day setting, outpatient, or ongoing hospitalization), and the recordings were analyzed to measure gastric retention, gastric transit time, and the completeness of the examinations. RESULTS: CE examination was performed in a hospital day setting in 100 patients, in an outpatient setting in 61, and during hospitalization in 29. Gastric retention of the capsule occurred in 8 of 190 patients (4.2%) (5 women, 3 men; mean age, 37.9 years). Gastric retention occurred during hospitalization in 7/29 (24.1%) and in hospital day setting in one of 100. Ongoing hospitalization was statistically associated with an increased risk of gastric capsule retention (p < 0.0001). The cecum was reached in 165/190 (87%), and ongoing hospitalization was associated with a lower rate of complete examinations (p < 0.001). Small-bowel transit time was similar, regardless of the circumstances of CE performance. CONCLUSIONS: Ongoing hospitalization is a major cause of gastric retention and incomplete examination by CE. Therefore, CE performance during ongoing hospitalization may require the use of a prokinetic agent. PMID- 16246701 TI - A pilot study of endoscopic closure of colonic perforations with endoclips in a swine model. AB - BACKGROUND: Surgical closure of a colon perforation is accompanied by the risks of general anesthesia and prolonged recovery from surgery because of ileus and other sequelae. Very little is known about the effectiveness of endoluminal repair of colon perforations with clips, which eliminates incisions of the abdominal wall and provides a less invasive alternative to surgical closure. The aim of this study is to evaluate the feasibility and the safety of endoscopic closure of colonic perforations with endoclips in a porcine model. METHODS: Approximately 1.5- to 2-cm colon perforations created with a needle knife in 4 50 kg, female pigs that were under general anesthesia were closed with endoclips. After 24 hours of recovery, the animals were allowed to eat. All the animals received intravenous antibiotics and were carefully monitored for signs of sepsis. After a follow-up of 1 week, the pigs were euthanized for postmortem examination. The fifth pig was euthanized immediately after closure of a 5-cm colon perforation with clips to evaluate the extent of transmural closure with endoclips. RESULTS: The animals recovered well, without any clinical features of sepsis or peritonitis. Postmortem examination did not reveal fecal peritonitis, and there was no evidence of pericolonic abscess formation at the site of perforation. The perforation site showed signs of healing without any evidence of transmural dehiscence. Histopathology demonstrated granulation tissue bridging the site of perforation. In the fifth pig, euthanized immediately after closure of the perforation, nice mucosal apposition was seen, while the muscular and serosal coats remained dehisced. CONCLUSIONS: Endoscopic closure of small iatrogenic colon perforations with clips results in mucosal and submucosal healing and prevents fecal soiling of peritoneal cavity. PMID- 16246702 TI - Video capsule endoscopy and celiac disease. PMID- 16246703 TI - Capsule endoscopy in celiac disease. PMID- 16246704 TI - Tuberculous mesenteric lymphadenitis involving the gastric wall: case report. PMID- 16246705 TI - Complications after traction removal of direct percutaneous endoscopic jejunostomy: three case reports. PMID- 16246707 TI - "Hot stuff": EUS-guided brachytherapy. PMID- 16246706 TI - EUS-guided brachytherapy. PMID- 16246708 TI - Double endoscope method to access intradiverticular papilla. PMID- 16246709 TI - Heterotopic pancreas in the esophagus diagnosed by EUS-guided FNA. PMID- 16246710 TI - Metastatic low-grade endometrial stromal sarcoma presented as a subepithelial mass in the stomach was diagnosed by EUS-guided FNA. PMID- 16246711 TI - Endoscopic diverticulotomy with an isolated-tip papillotome (Iso-Tome) in a patient with intraluminal duodenal diverticulum. PMID- 16246712 TI - Photodynamic therapy is nonthermal injury. PMID- 16246714 TI - An overtube is not required for reducing post-PEG peristomal infection. PMID- 16246716 TI - Valved esophageal stents: benefits for patients outweigh perceived risks. PMID- 16246718 TI - From transcription to mRNA: PAF provides a new path. AB - Two new studies reveal a novel connection between transcription and pre-mRNA processing: the Paf1 elongation complex is involved in the 3' end formation of polyadenylated and nonpolyadenylated RNA polymerase II transcripts. PMID- 16246719 TI - E Pluribus Unum: 3' end formation of polyadenylated mRNAs, histone mRNAs, and U snRNAs. AB - The 3' ends of almost all eukaryotic RNAs are generated by nucleolytic cleavage. Remarkably, three groups now demonstrate that similar or identical endonucleases of the metallo-beta-lactamase family generate the 3' ends of polyadenylated mRNAs, nonpolyadenylated histone mRNAs, and U snRNAs. PMID- 16246720 TI - Molecular links between centrosome and midbody. AB - The terminal step in cytokinesis that severs a cell in two-abscission-is poorly understood. In Developmental Cell, Fabbro et al (2005) identify a centrosome protein whose multiple phosphorylations regulate its movement from centrosome to midbody and completion of abscission. PMID- 16246721 TI - The nucleation and maintenance of heterochromatin by a histone deacetylase in fission yeast. AB - Posttranslational modifications of histones play an essential role in heterochromatin assembly. Whereas the role of Clr4/Suv39h-mediated methylation of histone H3 at lysine 9 (H3K9) in heterochromatin assembly is well studied, the exact function of histone deacetylases (HDACs) in this process is unclear. We show that Clr3, a fission yeast homolog of mammalian class II HDACs, acts in a distinct pathway parallel to RNAi-directed heterochromatin nucleation to recruit Clr4 and mediate H3K9 methylation at the silent mating-type region and centromeres. At the mat locus, Clr3 is recruited at a specific site through a mechanism involving ATF/CREB family proteins. Once recruited, Clr3 spreads across the 20 kb silenced domain that requires its own HDAC activity and heterochromatin proteins including Swi6/HP1. We also demonstrate that Clr3 contributes to heterochromatin maintenance by stabilizing H3K9 trimethylation and by preventing histone modifications associated with active transcription, and that it limits RNA polymerase II accessibility to naturally silenced repeats at heterochromatin domains. PMID- 16246722 TI - Recognition of RNA polymerase II and transcription bubbles by XPG, CSB, and TFIIH: insights for transcription-coupled repair and Cockayne Syndrome. AB - Loss of a nonenzymatic function of XPG results in defective transcription-coupled repair (TCR), Cockayne syndrome (CS), and early death, but the molecular basis for these phenotypes is unknown. Mutation of CSB, CSA, or the TFIIH helicases XPB and XPD can also cause defective TCR and CS. We show that XPG interacts with elongating RNA polymerase II (RNAPII) in the cell and binds stalled RNAPII ternary complexes in vitro both independently and cooperatively with CSB. XPG binds transcription-sized DNA bubbles through two domains not required for incision and functionally interacts with CSB on these bubbles to stimulate its ATPase activity. Bound RNAPII blocks bubble incision by XPG, but an ATP hydrolysis-dependent process involving TFIIH creates access to the junction, allowing incision. Together, these results implicate coordinated recognition of stalled transcription by XPG and CSB in TCR initiation and suggest that TFIIH dependent remodeling of stalled RNAPII without release may be sufficient to allow repair. PMID- 16246723 TI - Molecular basis for the recognition of phosphorylated and phosphoacetylated histone h3 by 14-3-3. AB - Phosphorylation of histone H3 is implicated in transcriptional activation and chromosome condensation, but its immediate molecular function has remained obscure. By affinity chromatography of nuclear extracts against modified H3 tail peptides, we identified 14-3-3 isoforms as proteins that bind these tails in a strictly phosphorylation-dependent manner. Acetylation of lysines 9 and 14 does not impede 14-3-3 binding to serine 10-phosphorylated H3 tails. In vivo, 14-3-3 is inducibly recruited to c-fos and c-jun nucleosomes upon gene activation, concomitant with H3 phosphoacetylation. We have determined the structures of 14-3 3zeta complexed with serine 10-phosphorylated or phosphoacetylated H3 peptides. These reveal a distinct mode of 14-3-3/phosphopeptide binding and provide a structural understanding for the lack of effect of acetylation at lysines 9 and 14 on this interaction. 14-3-3 isoforms thus represent a class of proteins that mediate the effect of histone phosphorylation at inducible genes. PMID- 16246724 TI - A posttranscriptional role for the yeast Paf1-RNA polymerase II complex is revealed by identification of primary targets. AB - The yeast Paf1 complex (Paf1C: Paf1, Cdc73, Ctr9, Rtf1, and Leo1) is associated with RNA Polymerase II (Pol II) at promoters and coding regions of transcriptionally active genes, but transcript abundance for only a small subset of genes is altered by loss of Paf1. By using conditional and null alleles of PAF1 and microarrays, we determined the identity of both primary and secondary targets of the Paf1C. Neither primary nor secondary Paf1C target promoters were responsive to loss of Paf1. Instead, Paf1 loss altered poly(A) site utilization of primary target genes SDA1 and MAK21, resulting in increased abundance of 3' extended mRNAs. The 3'-extended MAK21 RNA is sensitive to nonsense-mediated decay (NMD), as revealed by its increased abundance in the absence of Upf1. Therefore, although the Paf1C is associated with Pol II at initiation and during elongation, these critical Paf1-dependent changes in transcript abundance are due to alterations in posttranscriptional processing. PMID- 16246725 TI - A Requirement for the Saccharomyces cerevisiae Paf1 complex in snoRNA 3' end formation. AB - RNA synthesis and processing are coordinated by proteins that associate with RNA polymerase II (pol II) during transcription elongation. The yeast Paf1 complex interacts with RNA pol II and mediates histone modifications during elongation. To elucidate the functions of this complex, we isolated missense mutations in the gene encoding the Rtf1 subunit and used them to identify functionally interacting proteins. We identified NAB3 as a dosage suppressor of rtf1. Nab3, together with Nrd1, directs 3' end formation of nonpolyadenylated RNA pol II transcripts, such as snoRNAs. Deletion of Paf1, but not the Set1, Set2, or Dot1 histone methyltransferases, causes accumulation of snoRNA transcripts that are extended at their 3' ends. The Paf1 complex associates with and facilitates Nrd1 recruitment to the SNR47 gene, suggesting a direct involvement in 3' end formation. Our results reveal a posttranscriptional function for the Paf1 complex, which appears unrelated to its role in histone methylation. PMID- 16246726 TI - The GIT-associated kinase PAK targets to the centrosome and regulates Aurora-A. AB - Previously, we showed PAK-PIX-GIT targets and regulates focal adhesions; here, we uncover a different function for the complex at the centrosome. Active PAK1 is particularly evident in mitosis and phosphorylates the centrosomal adaptor GIT1 on serine 517. Interestingly, direct centrosome targeting activates the kinase via a process not requiring Rho GTPases; excision of the centrosome prevents this activation. Once activated, PAK1 dissociates from PIX/GIT but can bind to and phosphorylate the important centrosomal kinase Aurora-A. PAK1 promotes phosphorylation of Aurora-A on Thr288 and Ser342, which are key sites for kinase activation in mitosis. In vivo PAK activation causes an accumulation of activated Aurora-A; conversely, when betaPIX is depleted or PAK is inhibited, there is a delay in centrosome maturation. These observations may underlie reported effects of active PAK on cells, including histone H3 phosphorylation, alterations in centrosome number, and progression through mitosis. PMID- 16246727 TI - Pi release from eIF2, not GTP hydrolysis, is the step controlled by start-site selection during eukaryotic translation initiation. AB - Irreversible GTP hydrolysis by eIF2 is a critical step in translation initiation in eukaryotes because it is thought to commit the translational machinery to assembling the ribosomal complex at the selected point in the mRNA. Our quantitative analysis of the steps and interactions involved in activating GTP hydrolysis by eIF2 during translation initiation in vitro indicates that a structural rearrangement in the 43S preinitiation complex activates it to become fully competent to hydrolyze GTP. Contrary to the prevailing model, release of inorganic phosphate after GTP hydrolysis by eIF2, not hydrolysis itself, is controlled by recognition of the AUG codon. Release of P(i), which makes GTP hydrolysis irreversible, appears to be controlled by the AUG-dependent dissociation of eIF1 from the preinitiation complex. PMID- 16246728 TI - Roles of eukaryotic ribosomal proteins in maturation and transport of pre-18S rRNA and ribosome function. AB - Despite the rising knowledge about ribosome function and structure and how ribosomal subunits assemble in vitro in bacteria, the in vivo role of many ribosomal proteins remains obscure both in pro- and eukaryotes. Our systematic analysis of yeast ribosomal proteins (r-proteins) of the small subunit revealed that most eukaryotic r-proteins fulfill different roles in ribosome biogenesis, making them indispensable for growth. Different r-proteins control distinct steps of nuclear and cytoplasmic pre-18S rRNA processing and, thus, ensure that only properly assembled ribosomes become engaged in translation. Comparative analysis of dynamic and steady-state maturation assays revealed that several r-proteins are required for efficient nuclear export of pre-18S rRNA, suggesting that they form an interaction platform with the export machinery. In contrast, the presence of other r-proteins is mainly required before nuclear export is initiated. Our studies draw a correlation between the in vitro assembly, structural localization, and in vivo function of r-proteins. PMID- 16246729 TI - Folding of CFTR is predominantly cotranslational. AB - The folding process for newly synthesized, multispanning membrane proteins in the endoplasmic reticulum (ER) is largely unknown. Here, we describe early folding events of the cystic fibrosis transmembrane conductance regulator (CFTR), a member of the ABC-transporter family. In vitro translation of CFTR in the presence of semipermeabilized cells allowed us to investigate this protein during nascent chain elongation. We found that CFTR folds mostly during synthesis as determined by protease susceptibility. C-terminally truncated constructs showed that individual CFTR domains formed well-defined structures independent of C terminal parts. We conclude that the multidomain protein CFTR folds mostly cotranslationally, domain by domain. PMID- 16246730 TI - ERp29 triggers a conformational change in polyomavirus to stimulate membrane binding. AB - Membrane penetration of nonenveloped viruses is a poorly understood process. We have investigated early stages of this process by studying the conformational change experienced by polyomavirus (Py) in the lumen of the endoplasmic reticulum (ER), a step that precedes its transport into the cytosol. We show that a PDI like protein, ERp29, exposes the C-terminal arm of Py's VP1 protein, leading to formation of a hydrophobic particle that binds to a lipid bilayer; this reaction likely mimics initiation of Py penetration across the ER membrane. Expression of a dominant-negative ERp29 decreases Py infection, indicating ERp29 facilitates viral infection. Interestingly, cholera toxin, another toxic agent that crosses the ER membrane into the cytosol, is unfolded by PDI in the ER. Our data thus identify an ER factor that mediates membrane penetration of a nonenveloped virus and suggest that PDI family members are generally involved in ER remodeling reactions. PMID- 16246731 TI - Lysine 63 polyubiquitination of the nerve growth factor receptor TrkA directs internalization and signaling. AB - Nerve growth factor (NGF) binding to p75(NTR) influences TrkA signaling, yet the molecular mechanism is unknown. We observe that NGF stimulates TrkA polyubiquitination, which was attenuated in p75(-/-) mouse brain. TrkA is a substrate of tumor necrosis factor receptor-associated factor 6 (TRAF6), and expression of K63R mutant ubiquitin or an absence of TRAF6 abrogated TrkA polyubiquitination and internalization. NGF stimulated formation of a TrkA/p75(NTR) complex through the p62 scaffold, recruiting the E3/TRAF6 and E2/UbcH7. Peptide targeted to the TRAF6 binding site present in p62 blocked interaction with TRAF6 and inhibited ubiquitination of TrkA, signaling, internalization, and NGF-dependent neurite outgrowth. Mutation of K485 to R blocked TRAF6 and NGF-dependent polyubiquitination of TrkA, resulting in retention of the receptor on the membrane and an absence in activation of specific signaling pathways. These findings reveal that polyubiquitination serves as a common platform for the control of receptor internalization and signaling. PMID- 16246732 TI - An electrostatic steering mechanism of Cdc42 recognition by Wiskott-Aldrich syndrome proteins. AB - The specific and rapid formation of protein complexes is essential for diverse cellular processes such as remodeling of actin filaments in response to the interaction between Rho GTPases and the Wiskott-Aldrich syndrome proteins (WASp and N-WASp). Although Cdc42, TC10, and other members of the Rho family have been implicated in binding to and activating the WAS proteins, the exact nature of such a protein-protein recognition process has remained obscure. Here, we describe a mechanism that ensures rapid and selective long-range Cdc42-WASp recognition. The crystal structure of TC10, together with mutational and bioinformatic analyses, proved that the basic region of WASp and two unique glutamates in Cdc42 generate favorable electrostatic steering forces that control the accelerated WASp-Cdc42 association reaction. This process is a prerequisite for WASp activation and a critical step in temporal regulation and integration of WASp-mediated cellular responses. PMID- 16246733 TI - Structural basis for inhibition of the insulin receptor by the adaptor protein Grb14. AB - Grb14, a member of the Grb7 adaptor protein family, possesses a pleckstrin homology (PH) domain, a C-terminal Src homology-2 (SH2) domain, and an intervening stretch of approximately 45 residues known as the BPS region, which is unique to this adaptor family. Previous studies have demonstrated that Grb14 is a tissue-specific negative regulator of insulin receptor signaling and that inhibition is mediated by the BPS region. We have determined the crystal structure of the Grb14 BPS region in complex with the tyrosine kinase domain of the insulin receptor. The structure reveals that the N-terminal portion of the BPS region binds as a pseudosubstrate inhibitor in the substrate peptide binding groove of the kinase. Together with the crystal structure of the SH2 domain, we present a model for the interaction of Grb14 with the insulin receptor, which indicates how Grb14 functions as a selective protein inhibitor of insulin signaling. PMID- 16246734 TI - Neural substrates for verbal working memory in deaf signers: fMRI study and lesion case report. AB - The nature of the representations maintained in verbal working memory is a topic of debate. Some authors argue for a modality-dependent code, tied to particular sensory or motor systems. Others argue for a modality-neutral code. Sign language affords a unique perspective because it factors out the effects of modality. In an fMRI experiment, deaf participants viewed and covertly rehearsed strings of non-sense signs; analyses focused on regions responsive in both sensory and rehearsal phases. Compared with previous findings in hearing subjects, deaf subjects showed a significantly increased involvement of parietal regions. A lesion case study indicates that this network is left-dominant. The findings support the hypothesis that linguistic working memory is supported by modality specific neural systems, but some modality-neutral systems may also be involved. PMID- 16246735 TI - Impaired verb fluency: a sign of mild cognitive impairment. AB - We assessed verb fluency vs. noun and letter-based fluency in 199 subjects referred for cognitive complaints including Subjective Cognitive Impairment, Mild Cognitive Impairment, and Alzheimer's disease. ANCOVAs and factor analyses identified verb, noun, and letter-based fluency as distinct tasks. Verb fluency performance in Mild Cognitive Impairment differed significantly from Subjective Cognitive Impairment and Alzheimer's disease. Reduced verb fluency thus appears to be a linguistic marker for incipient dementia. One possibility is that the verb fluency deficit in Mild Cognitive Impairment results from degenerative processes known to occur in the parahippocampal region. PMID- 16246736 TI - Syllable structure and sonority in language inventory and aphasic neologisms. AB - Phonological theories have raised the notion of a universally preferred syllable type which is defined in terms of its sonority structure (e.g., ). Empirical evidence for this notion has been provided by distributional analyses of natural languages and of language acquisition data, and by aphasic speech error analyses. The present study investigates frequency distributions of syllable types in German, which allows for a rather complex syllable structure, and in neologistic utterances of a German speaking jargon aphasic. The findings suggest that the sonority structure of the patient's neologisms is generally in accordance with the notion of theoretically preferred syllables. Moreover, comparative analyses suggest that the predominance of the preferred syllable type is especially pronounced in the aphasic data. On the basis of these findings, the influence of sonority in impaired phonological lexical processing is discussed. PMID- 16246737 TI - Grammatical category ambiguity in aphasia. AB - This study asked whether aphasic adults show different noun/verb retrieval patterns based upon their clinical categorization as fluent or nonfluent. Participants selected either the noun or the verb meaning of target words, as presented in three contexts. The framework was that nouns (associated with temporal lobe function) are processed, stored, and retrieved separately from verbs (associated with frontal lobe function), implying separate status in the mental lexicon. Stimuli were homophonic homographs, words that are spelled and pronounced the same but which have different meanings (in this case, noun and verb meanings). Another contrast was the putative difference between systematic pairs (e.g., "kiss" and "farm"), in which noun and verb meanings are transparently related, and may be stored as a unit, and unsystematic pairs (e.g., "squash" and "sink"), in which noun and verb meanings are apparently unrelated, implying discrete storage. Results demonstrated significant interactions between fluent and nonfluent participants, suggesting that, as expected, fluent aphasic adults have more difficulty with nouns, nonfluent aphasic adults have more difficulty with verbs. There was no effect of systematicity. Contrary to expectations, verbs proved less vulnerable, rather than more vulnerable, to aphasic impairment. PMID- 16246739 TI - The processing of the right-sided accent mark in left neglect dyslexia. AB - Italian polysyllabic words with stress falling on the last syllable are written with a diacritic sign on the last vowel. It allows discrimination between two words with the same orthographic segments (e.g., papa [pope], papa [dad]). The effect of the accent mark in left neglect dyslexia has never been investigated. In the current study, six patients with neglect dyslexia were asked to read aloud homographic words and non-words, with or without the accent mark. The presence of the accent improved reading performance of three patients but only with real words. In contrast, two patients ignored the accent mark, even if it relied on the right ipsilesional space. We conclude that the accent mark is computed separately from letter identity and that it can act as an important cue for lexical access. PMID- 16246740 TI - Contextual priming in semantic anomia: a case study. AB - The present case continues the series of anomia treatment studies with contextual priming (CP), being the second in-depth treatment study conducted for an individual suffering from semantically based anomia. Our aim was to acquire further evidence of the facilitation and interference effects of the CP treatment on semantic anomia. Based on the results of the study of , our hypothesis before the treatment was that our participant would show short-term interference and at most modest and short-term benefit from treatment. To acquire such evidence would not only be important for the choice of anomia treatment methods in individual patients, but would also prompt further development of the CP method. The CP technique used for our participant included cycles of repeating and naming items in three contextual conditions (semantic, phonological, and unrelated). As predicted, the overall improvement of naming was modest and short-term. Interestingly, the contextual condition that corresponded with the nature of our patient's underlying naming deficit (semantic) elicited immediate interference in the form of contextual naming errors, as well as short-term improvement of naming. Based on this and a recent study by , it appears that despite short-term positive effects, in its current form the CP treatment is not sufficient for those aphasics who have a semantic deficit underlying their anomia. The possible mechanism and directions for future research are discussed. PMID- 16246738 TI - Reduced phonological similarity effects in patients with damage to the cerebellum. AB - Ten cerebellar patients were compared to 10 control subjects on a verbal working memory task in which the phonological similarity of the words to be remembered and their modality of presentation were manipulated. Cerebellar patients demonstrated a reduction of the phonological similarity effect relative to controls. Further, this reduction did not depend systematically upon the presentation modality. These results first document that qualitative differences in verbal working memory may be observed following cerebellar damage, indicating altered cognitive processing, even though behavioral output as measured by the digit span may be within normal limits. However, the results also present problems for the hypothesis that the cerebellar role is specifically associated with articulatory rehearsal as conceptualized in the Baddeley-Hitch model of working memory. PMID- 16246741 TI - Cerebral asymmetries in early orthographic and phonological reading processes: evidence from backward masking. AB - A lateralized backward masking paradigm was used to examine hemisphere differences in orthographic and phonological processes at an early time course of word recognition. Targets (e.g., bowl) were presented and backward masked by either pseudohomophones of the target word (orthographically and phonologically similar, e.g., BOAL), orthographically similar (little phonological similarity, e.g., BOOL), or unrelated (e.g., MANT) non-words. Stimuli were presented to the left visual field (LVF) or right visual field (RVF) using target/mask durations of 30 or 50 ms. Responses to RVF presentations were facilitated in the pseudohomophone condition relative to the orthographically similar condition. This provides evidence that the LH can access phonology in the earliest moments of word recognition. In contrast, responses to LVF stimuli showed a greater degree of facilitation for the orthographically similar condition relative to the unrelated condition as compared to RVF presentations. These results suggest the Strong Phonological Theory of Frost (1998) is potentially an accurate description of reading processes supported by the LH, but may not be applicable to the RH. The lexical route offered by the Dual Route Cascaded model may better describe reading processes at early moments in the RH (Coltheart, Rastle, Perry, Langdon, & Ziegler, 2001). PMID- 16246742 TI - Reaction time and accuracy in individuals with aphasia during auditory vigilance tasks. AB - Research indicates that attentional deficits exist in aphasic individuals. However, relatively little is known about auditory vigilance performance in individuals with aphasia. The current study explores reaction time (RT) and accuracy in 10 aphasic participants and 10 nonbrain-damaged controls during linguistic and nonlinguistic auditory vigilance tasks. Findings indicate that the aphasic group was less accurate during both tasks than the control group, but was not slower in their accurate responses. Further examination of the data revealed variability in the aphasic participants' RT contributing to the lower accuracy scores. PMID- 16246743 TI - Intensity-modulated radiotherapy as the boost or salvage treatment of nasopharyngeal carcinoma: the appropriate parameters in the inverse planning and the effect of patient's anatomic factors on the planning results. AB - The current study demonstrates that the large increase in normal tissue penalty often degrades target dose uniformity without a concomitant large improvement in normal tissue dose, especially in anatomically unfavorable patients. The excessively large normal tissue penalties do not improve treatment plans for patients having unfavorable geometry. PMID- 16246744 TI - Comparison of two dose calculation methods applied to extracranial stereotactic radiotherapy treatment planning. AB - BACKGROUND AND PURPOSE: Lung tissue is a special challenge for a dose calculation algorithm, especially in the case of extracranial stereotactic radiotherapy (ESRT) due to small field sizes in combination with large variations in tissue density. The present study investigates the choice of dose calculation algorithm for 18 patients with a single lung tumor and 8 patients with a single liver tumor. The dose calculation is performed with both the pencil beam convolution algorithm and the collapsed cone convolution algorithm with the same number of monitor units in both cases. In addition, the dose calculation with the collapsed cone convolution algorithm is also performed with modified field sizes in order to match the Planning Target Volume (PTV) peripheral dose of the pencil beam based treatment. RESULTS: For liver tumors, the mean Clinical Target Volume (CTV) dose calculated by the collapsed cone convolution algorithm and the pencil beam convolution algorithm is almost identical. For lung tumors, the mean CTV dose determined by the collapsed cone convolution algorithm differs up to 20%. Plans obtained by the two algorithms have field sizes which differ up to 8mm for the same number of monitor units and minimum dose to the lung PTV. CONCLUSIONS: The choice of dose calculation algorithm can have a large influence on a treatment plan for ESRT of the lungs. PMID- 16246745 TI - Truth or consequences. PMID- 16246746 TI - Late calcific deposition on the hydrophilic acrylic MemoryLens. PMID- 16246747 TI - Is it time to remove the MemoryLens IOL from the market? PMID- 16246748 TI - Accommodation measurements in prepresbyopic and presbyopic populations. PMID- 16246749 TI - Prevention of endophthalmitis. PMID- 16246752 TI - Insertion of the Sensar IOL via the Monarch II delivery system. PMID- 16246755 TI - Chopstick technique for nucleus removal in an impending dropped nucleus. PMID- 16246756 TI - Effect of brimonidine on pupil diameter. PMID- 16246758 TI - Chopstick technique for nucleus removal in an impending dropped nucleus. PMID- 16246759 TI - Late ptosis after laser in situ keratomileusis. PMID- 16246760 TI - Consultation section. Refractive surgical problem. PMID- 16246768 TI - Intraoperative management of iris prolapse using iris hooks. AB - Iris prolapse has become an unusual and generally benign event in routine cataract surgery using endocapsular phacoemulsification techniques. We operated on the second eye of a patient with hyperopia whose first eye surgery in our care had been uneventful. In the second eye, as soon as the phaco incision was made and hydrodissection begun through the primary incision, the iris repeatedly prolapsed through the incision. This was managed definitively by applying iris hooks, 1 at each side of the primary incision, to the pupillary border so the prolapsing iris was held peripherally in the anterior chamber and under modest tension. PMID- 16246769 TI - Illuminated endochopper in the management of posteriorly dislocated lens nucleus. AB - We describe the use of an illuminated endochopper (a prototype instrument produced by DORC International) in the management of a posteriorly dislocated lens nucleus or lens particles. This instrument helps to divide the lens nucleus or its fragments into small pieces and thus reduces time and ultrasound energy. PMID- 16246770 TI - Microkeratome-assisted phacoemulsification. AB - This study reports a technique in which phacoemulsification and intraocular lens (IOL) insertion are performed in conjunction with a microkeratome flap in a patient with ocular surface disease. Microkeratome-assisted phacoemulsification was performed in a 72-year-old woman with a history of trachoma during childhood. A 130 microm corneal flap was made using a microkeratome prior to phacoemulsification and IOL insertion. Lifting the flap during surgery allowed a clear view of the anterior chamber through the smooth lamellar interface created by the microkeratome blade. The flap was washed and repositioned without sutures at the end of surgery. There were no complications associated with the microkeratome flap following surgery. Visual acuity improved from hand motion to 20/200. Microkeratome-assisted phacoemulsification is a safe technique for cataract patients with diseases of the ocular surface. PMID- 16246771 TI - Spectrum of clear corneal incision cataract wound infection. AB - PURPOSE: To describe the presentation, management, and outcome of 5 patients who presented with main-port or side-port wound infection after uneventful clear corneal cataract surgery. SETTING: Ophthalmic Surgery Centre, Chatswood, and Department of Ophthalmology, Prince of Wales Hospital, Randwick, Australia, and Moorfields Eye Hospital NHS Foundation Trust, London, United Kingdom. METHODS: This retrospective case series comprised 5 patients who had uneventful clear corneal phacoemulsification surgery and developed either a main-port or side-port wound infection. The clinical features, microbiologic studies, management, and results are reported. RESULTS: The median age of the patients was 79 years. Infection of the main-port incision occurred in 3 cases and of the side port in 2 cases. The patients presented from a few days to several weeks after uneventful phacoemulsification. In 2 cases, the bacteria Pseudomonas aeruginosa and Staphylococcus aureus were isolated. In 1 case, the fungus Aspergillus was isolated and required extensive medical and surgical treatment. In the other 2 cases; empiric antimicrobial therapy was given because no organism was isolated in 1 case, and in the other milder case, microbiological investigations were not performed. Final visual acuity was 6/4 in 2 cases and 6/5, 6/12, and 6/18 in 1 case each. CONCLUSIONS: Bacterial or fungal wound infection can present within days or even several weeks following clear corneal cataract surgery. Patients with ocular discomfort or blurred vision after such surgery should be advised to report promptly. Rapid identification and appropriate management of patients with clear corneal wound infection can result in good visual outcomes. PMID- 16246772 TI - Brimonidine 0.15% versus apraclonidine 0.5% for prevention of intraocular pressure elevation after anterior segment laser surgery. AB - PURPOSE: To compare the efficacy and safety of brimonidine 0.15% with those of apraclonidine 0.5% in preventing intraocular pressure (IOP) elevations after anterior segment laser surgery. SETTING: Massachusetts Eye and Ear Infirmary, Glaucoma Service, Boston, Massachusetts, USA. METHODS: This double-masked randomized trial 80 eyes of 80 patients who had laser peripheral iridotomy, argon laser trabeculoplasty, or neodymium:YAG laser capsulotomy. Eyes received 1 drop of brimonidine 0.15% or apraclonidine 0.5% before laser surgery. Intraocular pressure, heart rate, and blood pressure were measured before laser surgery and at 1 hour, 3 hours, 24 hours, and 1 week after laser surgery. RESULTS: Before laser treatment, 41 patients received brimonidine 0.15% and 39 received apraclonidine 0.5%. Thirteen (31.7%) patients in the brimonidine group and 11 (28.2%) in the apraclonidine group had postoperative IOP elevations of 5 mm Hg or more (P = .5). Four patients (9.8%) in the brimonidine group and 3 (7.7%) in the apraclonidine group had IOP increases of 10 mm Hg or more (P = .5). There were no statistically significant changes in mean heart rate or blood pressure in either group except a slight reduction in diastolic blood pressure at 1 hour in the brimonidine group (-4.7 +/- 9.2 mm Hg) compared with that in the apraclonidine group (-0.1 +/- 9.1 mm Hg) (P = .01). No clinically significant side effects were noted in either group. CONCLUSION: A single preoperative drop of brimonidine 0.15% had similar efficacy and safety as apraclonidine 0.5% in preventing IOP elevations immediately after anterior segment laser surgery. PMID- 16246773 TI - Magnetic resonance imaging and Orbscan assessment of the anterior chamber. AB - PURPOSE: To evaluate the correlation between white-to-white (WTW) distance as assessed by Orbscan II (Bausch & Lomb) and high-resolution magnetic resonance imaging (MRI) sulcus-sulcus (S-S) measures and to analyze the correlation between age and anterior chamber parameters. SETTING: Istituto per la Ricerca e la Cura del Cancro, Candiolo, Torino, Italy. METHODS: Eighty-eight patients had MRI with a 1.5 Tesla imager (GE Medical Systems) using a 3-inch circular coil. T1 weighted fast spin-echo scans were performed on the axial plane. Orbscan II was also used to measure WTW distance, K, anterior chamber depth (ACD), and lens curvature. One eye was randomly selected for analysis. Measures were compared using Spearman correlation, paired Student t test, and analysis of variance (ANOVA; post hoc: Scheffe). Intersession variability was performed in 10 patients. Ten MRI images were randomly selected and measured by 2 operators in a masked fashion (intrasession variability). RESULTS: Intersession and intrasession correlation was good for MRI (r = 0.89 and r = 0.92, respectively), and intersession was good for Orbscan (r = 0.91). When comparing Orbscan II and MRI results, the ACD was well correlated and not significantly different, whereas the S-S and the WTW measures were not correlated and significantly different. Lens thickness and lens diameter were directly correlated with age; S-S, WTW, lens curvature, and ACD were inversely correlated with age. CONCLUSIONS: Current methods of measuring the WTW were poorly correlated with anatomical measures. The S-S diameter and other anterior chamber structures significantly change with age, which could give rise to potential problems with posterior chamber phakic intraocular lens sizing position. PMID- 16246774 TI - Measurement of the spatial shift of the pupil center. AB - PURPOSE: To evaluate the effectiveness of the pupil center as an anatomic landmark for excimer laser treatments. SETTING: Sekal-Microchirurgia-Rovigo Centre, Rovigo, Italy. METHODS: Pupillometry with the Costruzione Strumenti Oftalmici S.R.L. (CSO) pupil-measuring module (incorporated in Eye Top videokeratoscope) was performed in 52 patients with a diagnosis of myopia and in 25 patients with a diagnosis of hyperopia. Measurements both in mesopic and photopic conditions consisted of pupil diameters, spatial shift of the pupil center, and the distance between the pupil center and keratoscopic axis. RESULTS: The mean pupil diameter in photopic conditions of illumination in myopic eyes was 3.52 mm +/- 0.56 (SD), while in mesopic conditions it was 5.37 +/- 0.78 mm; in hyperopic eyes the mean photopic pupil diameter was 3.01 +/- 0.46 mm, while the mean mesopic diameter was 5.12 +/- 0.48 mm. The mean spatial shift of the pupil center in myopic eyes was 0.086 mm (maximum 0.269 mm), while in the hyperopic eyes it was 0.095 mm (maximum 0.283 mm). The mean distance between the pupil center and keratoscopic axis in myopic eyes was 0.226 +/- 0.13 mm (maximum 0.75 mm), while in hyperopic eyes it was 0.45 +/- 0.19 mm (maximum 0.8 mm). CONCLUSIONS: The mean of the measured pupil sizes was greater in myopic eyes than in hyperopic eyes. The spatial shift of the pupil center, as the pupil dilates, was relatively small in all groups; therefore, the pupil center is a good anatomic landmark for both traditional refractive surgery and wavefront-guided treatments. The mean distance between the keratoscopic axis and pupil center was greater in the hyperopic group than in the myopic group. Therefore, centration of any laser treatment on the basis of the keratoscopic analysis should be done carefully, especially in hyperopic eyes and in cases in which the pupil center is meaningfully shifted from keratoscopic axis, even in photopic conditions of illumination. PMID- 16246775 TI - Measurement of the internal diameter and depth of the anterior chamber: IOLMaster versus anterior chamber optical coherence tomographer. AB - PURPOSE: To study the internal diameter of the anterior chamber (AC), which remains a relatively subjective procedure dominated by the white-to-white technique, evaluated with a compass or using anterior segment photographs. In vivo dimensions of the AC were studied with an optical coherence tomographer (AC OCT) prototype. SETTING: Clinique Monticelli, Marseilles, France. METHODS: The dimensions in 107 normal eyes that had never had surgery were studied with the IOLMaster and AC OCT. RESULTS: The mean anterior chamber depth was 3.53 mm +/- 0.35 (SD) with the IOLMaster (Carl Zeiss, Medictec) and 3.64 +/- 0.33 mm with the AC OCT (Carl Zeiss, Medictec). The mean horizontal diameter of the AC was 12.12 +/- 0.44 mm with the IOLMaster and 12.20 +/- 0.44 mm with the AC OCT. With the AC OCT, the anterior chamber's horizontal and vertical diameter could be evaluated; in 74% of the observed eyes, the vertical diameter was larger by at least 100 mum than the horizontal diameter. DISCUSSION: The AC OCT has better reproducibility than the IOLMaster for evaluating the depth or internal diameter of the anterior chamber. This study showed that in 74% of cases, the AC was an oval with a larger vertical axis, which provides evidence of the value of the OCT preoperative examinations for phakic intraocular lens implantation. PMID- 16246776 TI - Central corneal thickness measurement with the Pentacam Scheimpflug system, optical low-coherence reflectometry pachymeter, and ultrasound pachymetry. AB - PURPOSE: To assess the intraoperator repeatability and interoperator reproducibility of central corneal thickness measurements by the Pentacam Scheimpflug imaging system (Oculus) and the optical low-coherence reflectometer (OLCR) pachymeter (Haag-Streit) and to compare them with those of ultrasound (US) pachymetry. SETTING: Assaf Harofe Medical Center Ophthalmology Outpatient Clinic, Zerifin, Israel. METHODS: Repeatability was determined from 10 successive measurements in each of 4 healthy patients. Reproducibility for the Pentacam Scheimpflug system was determined from measurements by 2 operators in each of 24 patients; in these 24 patients, central corneal thickness measurements were compared between the Pentacam and US pachymetry. For the OLCR pachymeter, reproducibility was determined from measurements by 2 operators in each of 16 patients, in whom central corneal thickness was also measured with the Pentacam. RESULTS: Mean coefficient of repeatability was 0.84% for the Pentacam Scheimpflug system and 0.33% for the OLCR pachymeter. For the Pentacam, the coefficient of interoperator reproducibility was 1.10% and the 95% limits of agreement were 10.2 microm to +11.9 microm. Mean difference between Pentacam and US was 6.09 microm. For the OLCR pachymeter, the coefficient of interoperator reproducibility was 0.59% and the 95% limits of agreement were -5.4 microm to +7.0 microm. Mean difference between central corneal thickness values obtained with the OLCR pachymeter and Pentacam Scheimpflug system was 1.7 microm. CONCLUSIONS: Objective, noncontact measurement of central corneal thickness with the Pentacam Scheimpflug system and OLCR pachymeter was convenient and yielded excellent intraoperator repeatability and interoperator reproducibility. Central corneal thickness values obtained with the Pentacam were similar to those obtained with both the OLCR pachymeter and an US pachymeter. Further research is needed to corroborate whether central corneal thickness measurements by the Pentacam and OLCR devices can be used interchangeably and are more clinically useful than US pachymetry. PMID- 16246777 TI - Comparison of contrast sensitivity and color discrimination after clear and yellow intraocular lens implantation. AB - PURPOSE: To compare contrast sensitivity and color vision in patients in whom blue-light filtering and non-yellow-tinted intraocular lenses (IOLs) were implanted. SETTING: Refractive Surgery Unit, Hospital NISA Valencia al Mar, Valencia, Spain. METHODS: Forty eyes of 20 patients were enrolled in a blue-light filtering fellow-eye control study; patients were implanted with a yellow-tinted IOL (AcrySof Natural, Alcon) in 1 eye and a non-yellow-tinted IOL (AcrySof SA60AT, Alcon) in the fellow eye after cataract surgery. Three months postoperatively, monocular contrast sensitivity function was measured with the CSV 1000-E contrast sensitivity chart at distance and the color discrimination with the Farnsworth-Munsell 100 Hue test. RESULTS: Eyes implanted with blue-light filtering IOLs showed similar contrast sensitivity to that in fellow eyes implanted with non-yellow-tinted IOLs (P>.1). Both types of IOLs showed normal contrast sensitivity values (normalized log-contrast sensitivity about 1.0). There were no statistically significant differences in chromatic discrimination between the 2 types of IOLs (P = .56). CONCLUSION: The use of blue-light filtering IOLs is more advisable because they are capable of protecting the retina against ultraviolet light without disturbance of contrast sensitivity and chromatic vision, which produces subjective impairment in visual function. PMID- 16246778 TI - Lipid tear deficiency in persistent dry eye after laser in situ keratomileusis and treatment results of new eye-warming device. AB - PURPOSE: To determine whether lipid tear deficiency is a significant pathogenic factor in persistent dry eyes after laser in situ keratomileusis (LASIK). SETTING: Ocular Surface Center, Miami, Florida, USA. METHODS: Thirty-four eyes of 17 patients (mean 46.35 years +/- 11 [SD]) complaining of persistent dryness more than 12 months after LASIK were prospectively studied by symptom scoring and kinetic analysis of tear interference image, tear breakup time, and fluorescein clearance test. Once patients had been clear of inflammation and treated for aqueous tear deficiency, lipid tear deficiency was further confirmed and treated with Eyefeel (Kao, Inc.), an eye-warming device, 4 times daily for 4 weeks. RESULTS: Sixteen patients were asymptomatic before LASIK but dryness persisted for 41 +/- 19.3 months. Delayed tear clearance was observed in 15 patients (88.2%) and floppy lids in 12 patients (70.5%). Aqueous tear deficiency was reconfirmed in 16 eyes (53.3%). After Eyefeel treatment, there was a subjective improvement of ocular surface diseases index from 60.6 +/- 10.6 to 25.8 +/- 18.5 (P = .0007). Tear breakup time was improved from 2.4 +/- 3.9 seconds to 7.9 +/- 3.6 seconds (P = .004). There was a tear interference pattern change from a vertical lipid tear deficiency to a horizontal normal in 7 eyes. There was a mean lipid spread time improvement from 1.3 +/- 0.4 sec to 0.8 +/- 0.4 sec (P = .001), and there was a mean lipid thickness improvement from 63.5 +/- 23 nm to 79.5 +/- 27 nm (P = .04). CONCLUSION: Persistent nature of dry eye after LASIK is attributed to in part to delayed tear clearance, undercorrected aqueous tear deficiency, and nonrecognized lipid tear deficiency. PMID- 16246779 TI - Artificial iris-lens diaphragm in reconstructive surgery for aniridia and aphakia. AB - PURPOSE: To determine the efficacy and safety of surgical implantation of artificial iris-lens diaphragm in patients with anatomic or functional iris deficiencies, aphakia or cataract. SETTING: Svyatoslav Fyodorov MNTK Eye Microsurgery, Cheboksary, Russia. METHODS: Twenty eyes of 19 consecutive patients with combined iris and lens pathology of traumatic or congenital etiology were identified for an interventional noncomparative case series. The newly proposed model of an elastic iris-lens diaphragm with a colored haptic and additional support elements was implanted using various fixation approaches. RESULTS: Fifteen eyes (75%) experienced improvement in corrected visual acuities. The best spectacle-corrected visual acuity (BSCVA) in 2 eyes did not change, while the uncorrected visual acuity (UCVA) in these eyes increased. There were 3 eyes in which BSCVA deteriorated with no change or even slight improvement in UCVA. All patients were satisfied with the cosmetic results of the surgery and reported a decrease in glare and photophobia. There was 1 intraoperative complication of vitreous hemorrhage. Postoperatively, 2 cases of hyphema, 1 case of ciliochoroidal detachment, 4 eyes with exaggerated immediate postoperative reaction, and 1 eye with persistent low-grade cyclitis were observed. In 1 eye, there was persistent intraocular pressure rise. One eye showed signs of cystoid macular edema. No iris-lens diaphragm decentrations and no new or extensions of old retinal detachments were seen during the follow-up period. CONCLUSIONS: Artificial iris-lens diaphragm implantation effectively improved postoperative outcomes by correcting aphakia, reducing glare disability, and addressing cosmetic issues faced by iris-deficient, and aphakic or cataract patients. Although the iris-lens diaphragm appears to be safe, long-term results must be clarified in studies with longer follow-up and a larger patient population. PMID- 16246780 TI - Patient pain during stretching of small pupils in phacoemulsification performed using topical anesthesia. AB - PURPOSE: To assess the pain experienced by patients with small pupils during pupil stretching in phacoemulsification performed using topical anesthesia. SETTING: Royal Victoria Eye & Ear Hospital and Mater Misericordiae University Hospital, Dublin, Ireland. METHODS: This was a prospective study that included 26 eyes with small pupils requiring mechanical pupil stretching during phacoemulsification cataract surgery performed under topical anesthesia without sedation. RESULTS: The mean pain score for the instillation of anesthetic drops (2.02) was higher than the mean pain score for the pupil stretch (1.63), but this difference was not significant (signed rank test = -32; P = .2738). There was no significant correlation between the duration of surgery and the overall pain score (r = 0.345; P = .08). There was no significant correlation between change in pupil size and either the pupil stretch score (r = -0.069; P = .74) or the overall pain score (r = -0.032; P = .8739). CONCLUSIONS: Pupil stretching during phacoemulsification in patients with small pupils was performed with minimal patient-reported pain using topical anesthesia. Stretching small pupils with a mechanical device during phacoemulsification performed under topical anesthesia was a safe procedure and did not result in significant patient discomfort. PMID- 16246781 TI - Pharmacological management of night vision disturbances after refractive surgery Results of a randomized clinical trial. AB - PURPOSE: To evaluate the efficacy and safety of diluted aceclidine eyedrops in reducing night vision disturbances after refractive surgery. SETTING: Department of Ophthalmology, Istituto Clinico Humanitas, Rozzano-Milano, Italy. METHODS: This double-masked randomized clinical trial included 30 patients (60 eyes) with chronic night vision disturbance after refractive surgery. Patients were randomly allocated to receive (1) placebo, (2) aceclidine 0.016%, or (3) aceclidine 0.032%. Drugs were administered once or twice daily. Anterior segment, haze, uncorrected visual acuity, best corrected visual acuity, intraocular pressure, corneal maps, and scotopic pupil size were determined at baseline and at follow up examinations (15 and 30 days after inclusion). Halos and double vision 4-step scales were built to determine subjective grading of night vision disturbance, and the root mean square (RMS) was calculated to determine objective changes in night vision disturbance. RESULTS: The effect of diluted aceclidine started about 15 minutes after instillation and lasted for about 5 hours. No difference between the 2 dilutions could be found. Thirty-nine of 40 treated eyes showed a reduction in night vision disturbance. The mean reduction in halos and double vision grading was 1.42 +/- 0.5 (SD) and 1.14 +/- 0.4, respectively. A mean decrease in pupil size of 2.5 mm was measured. Thirty minutes after the instillation of diluted aceclidine, the topography-derived wavefront error showed a statistically significant reduction in RMS values (total, spherical, astigmatic, coma, and higher order), which was maintained for 5 hours. A transitory conjunctival hyperemia was the only side effect reported. CONCLUSION: Diluted aceclidine seemed to be an effective and safe treatment for night vision disturbance following refractive surgery. PMID- 16246782 TI - In-use study of potential bacterial contamination of ophthalmic moxifloxacin. AB - PURPOSE: To determine the contamination rate of topical moxifloxacin 0.5% (Vigamox) after clinical use for preoperative and postoperative prophylaxis for cataract surgery. SETTING: Bascom Palmer Eye Institute, Miami, Florida, USA. METHODS: A total of 61 bottles of moxifloxacin were collected in a 2-month time period after use preoperatively by patients (n = 21, mean 2.2 days), postoperatively by patients (n = 16, mean 7.2 days), or by nurses in the operating room (n = 24). For each bottle, cultures of the cap, tip, external thread, and solution were inoculated onto chocolate agar plates and incubated at 35 degrees for 7 days. A total of 13 bottles of benzalkonium chloride (BAC) preserved gatifloxacin ophthalmic solution (Zymar) were also cultured in a similar manner for comparison. RESULTS: No organisms were found by cultures of the cap, tip, or solution from any bottle in the study. One colony of coagulase negative Staphylococcus was recovered from the thread of 1 Vigamox bottle. This bottle had negative cultures from its cap, tip, and solution. CONCLUSIONS: The only contaminant found in this study was cultured from the bottle thread. This is a location with a high rate of skin contact and minimal solution contact. The remaining cultures from this bottle were negative, showing the solution's ability to self-preserve. These data demonstrate that BAC-free Vigamox can be treated no differently than BAC-preserved solutions by patients and medical staff, without concern for solution contamination. PMID- 16246783 TI - Endoscope-assisted transscleral suture fixation to reduce the incidence of intraocular lens dislocation. AB - PURPOSE: To examine the surgical complications of transscleral sulcus suture of posterior chamber intraocular lenses (IOLs) and to compare the rates of surgical complications between patients in the nonendoscope-assisted and endoscope assisted groups. SETTING: Kyoto University Hospital, Kyoto, Japan. METHODS: This retrospective nonrandomized study comprised 121 eyes of 115 patients who had transscleral sulcus suture fixation of an IOL by the ab externo method. Typical 3 port vitrectomy was performed in 26 eyes in which the needle entry site and the haptic location were controlled using an endoscope. RESULTS: During follow-up of at least 3 months, surgical complications in the nonendoscope-assisted group (95 eyes) included IOL dislocation in 22 eyes (23%), high astigmatism in 12 eyes (13%), transient ocular hypertension in 10 eyes (11%), vitreous hemorrhage in 5 eyes (5.3%), retinal detachment in 4 eyes (4.2%), and cystoid macular edema in 2 eyes (2.0%). In the endoscope-assisted group (26 eyes), the same complications were markedly decreased: there were no cases of IOL dislocation, high astigmatism, vitreous hemorrhage, retinal detachment, or cystoid macular edema; transient ocular hypertension occurred in 1 eye (3.8%). There was a statistically significant difference in the incidence of IOL dislocation between the 2 groups (P<.01). CONCLUSION: Using an endoscope for transscleral sulcus suturing of an IOL can be an effective technique to reduce surgical complications, especially postoperative IOL dislocation. PMID- 16246784 TI - Topical anesthesia for transpupillary silicone oil removal combined with cataract surgery. AB - PURPOSE: To assess safety of topical anesthesia for transpupillary silicone oil removal in combination with cataract surgery. SETTING: Department of Ophthalmology Mannheim, University of Heidelberg, Mannheim, Germany. METHODS: The clinical interventional study included 37 consecutive patients having transpupillary silicone oil removal combined with cataract surgery. Without exception, surgery was carried out in topical anesthesia for all patients. During the study period, there were no patients having transpupillary silicone oil removal in another type of local anesthesia than topical anesthesia. Topical anesthesia was achieved with oxybuprocaine 0.4% eyedrops installed 4 to 5 times prior to surgery. Cataract surgery was performed using the clear cornea technique with implantation of a foldable intraocular posterior chamber lens. Silicone oil was released through a planned posterior capsulotomy during cataract surgery prior to implantation of the intraocular lens (IOL). RESULTS: For all patients, surgery could be carried out in topical anesthesia without switching to peribulbar or any other type of anesthesia. None of the patients complained about severe pain intraoperatively or postoperatively. No severe complications such as expulsive hemorrhage, luxation of the IOL, or iris incarceration were encountered in any of the surgeries. CONCLUSION: Transpupillary silicone oil through a planned posterior capsulotomy during cataract surgery may be performed in topical surgery. PMID- 16246785 TI - Combined clear corneal phacoemulsification and ab interno trabeculectomy: three year case series. AB - PURPOSE: To evaluate the efficacy and safety of a novel surgical approach, ab interno trabeculectomy, in a combined procedure (clear corneal phacoemulsification + ab interno trabeculectomy) for the management of concurrent cataract and glaucoma with prognostic factors for filtration failure. SETTING: Department of Ophthalmology, Palmanova Hospital, Udine, Italy. METHODS: A prospective noncomparative case series of 11 eyes of 11 consecutive cataract patients with medically uncontrolled primary open-angle glaucoma or pseudoexfoliation glaucoma was performed. Each patient had phacoemulsification + intraocular lens implantation + ab interno trabeculectomy, which consisted of a gonioscopically controlled ab interno removal of a quadrant (3 clock hours) of the trabecular meshwork. The main outcome measures were intraocular pressure (IOP), the number of antiglaucomatous medications used, and complications. RESULTS: Ten patients completed a 3-year follow-up. One patient had further surgery because of poor IOP control; in this case, the last valid observation was carried forward for IOP calculation. Mean preoperative IOP and IOPs measured 1, 3, 6, 12, 24, and 36 months after surgery were 25.0, 15.8, 15.4, 15.4, 15.2, 15.0, and 15.3 (-38.56%) mm Hg respectively. The number of medications averaged 2.4 before surgery and dropped to 0.8 at the end of follow-up. No major complications occurred during the follow-up period. CONCLUSIONS: The new surgical procedure combining phacoemulsification with ab interno trabeculectomy can induce a clinically relevant decrease in IOP in eyes with cataract and glaucoma with poor prognosis for filtering surgery. However, a randomized controlled clinical trial with a more extended follow-up and a larger series of patients is needed to ascertain the actual effectiveness and safety of this procedure. PMID- 16246786 TI - In vitro human corneal model to investigate stromal epithelial interactions following refractive surgery. AB - PURPOSE: To develop an in vitro human corneal model to evaluate stromal epithelial interactions following corneal refractive surgical procedures. SETTING: Department of Academic Ophthalmology, Rayne Institute, St. Thomas' Hospital, London, United Kingdom. METHODS: Fifty-six human donor corneas procured from the eye bank were placed in a specially designed acrylic corneal holder and were cultured using the air-interface organ culture technique for up to 4 weeks. Corneal refractive surgical procedures such as a simple epithelial defect, 4 diopter (D) and 9 D photorefractive keratectomy (PRK), 4 D and 9 D laser-assisted subepithelial keratectomy (LASEK), and 9 D laser in situ keratomileusis (LASIK) were performed on the model. Temporal events in epithelial and keratocyte cell kinetics were evaluated using digital imaging, confocal microscopy, and light microscopy. Two-way analysis of variance and Student t tests were used to assess statistical significance. RESULTS: Epithelial healing following PRK was completed by 92 hours +/- 10 (SD) at a rate of 0.58 +/- 0.45 mm2/hour. In LASEK, the epithelial flap was replaced by regenerating peripheral epithelium that showed significant delay in epithelial closure (120 +/- 5 hours) with prolonged latency (24 +/- 4 hours, P<.0001) in comparison with PRK. The magnitude of keratocyte loss corresponded to ablation depth, and keratocyte regeneration was dependent on epithelial closure. In comparison, LASIK corneas showed a lesser percentage of keratocyte loss with poor recovery of keratocyte density in the stromal flap. Epithelial viability and keratocyte density were well preserved in the in vitro human model as observed in control corneas for up to 4 weeks. CONCLUSIONS: The temporal events in stromal epithelial interactions in the in vitro human model closely mimicked in vivo observations. The human model further avoided species specific variations and provided a suitable test bed for evaluating newer algorithms and therapeutic regimens following refractive surgery. PMID- 16246787 TI - Feasibility and development of a high-power real accommodating intraocular lens. AB - PURPOSE: To develop an accommodating intraocular lens (IOL) that changes power with ciliary body action and generates a minimum of 8.0 diopters (D) by manipulation of a flexible material between a sulcus-fixated rigid plate and a ciliary muscle-operated capsular diaphragm. SETTING: Vissum-Instituto Oftalmologico de Alicante and Miguel Hernandez University, Alicante, Spain. METHODS: The feasibility of the concept was tested by a laboratory lens model. An implantable measuring device was constructed to simulate the lens action. The device was implanted in monkey eyes to measure the various parameters involved with such IOLs. Based on these measurements, an accommodating IOL prototype was built and implanted in monkeys' eyes. Pharmacologic agents were used to achieve ciliary relaxation and spasm. Ultrasound biomicroscopy (UBM) imaging was used to document the active changes of the IOL flexible lens curvature as related to the ciliary muscles status. RESULTS: The laboratory model produced more than 50.0 D of accommodation. The UBM demonstrated changes in lens curvature between cyclospasm to cycloplegia of calculated 9.0 to 53.0 D for flexible material with a refractive index of 1.41. CONCLUSIONS: Flexible material is capable of being manipulated when placed between a scleral-fixated rigid plane and the ciliary muscles-operated capsular diaphragm. Such manipulation as used by the IOL concept presented here has generated an active change of more than 40.0 D in the monkey eye. PMID- 16246788 TI - Optimal timing of capsular tension ring implantation: Miyake-Apple video analysis. AB - PURPOSE: To evaluate the appropriate timing for capsular tension ring implantation in cases of zonular weakness either prior to or after lens extraction using Miyake-Apple video analysis. SETTING: John Moran Eye Center, Salt Lake City, Utah, USA. METHODS: Four cadaver eyes were prepared using a standard Miyake-Apple protocol with image capture using digital video recording. After continuous curvilinear capsulorhexis and hydrodissection/viscodissection were performed, 2 eyes had early capsular tension ring implantation (CTR) and 2 eyes had CTR implantation after lens extraction. The 12.3 mm CTR was implanted in all eyes. Capsular bag torque and displacement, zonular elongation and stress, and ease of CTR placement were evaluated in each eye. RESULTS: Early CTR implantation resulted in significantly increased capsular torque and displacement of up to 4.0 mm compared to insertion in an empty capsular bag. There was significant zonular elongation and tension during early placement. CONCLUSION: In terms of minimizing further zonular stress and damage and capsular destabilization, the ideal timing for CTR placement is after lens extraction and decompression of the capsular bag. PMID- 16246789 TI - Effect of vancomycin, teicoplanin, and cefuroxime on Staphylococcus epidermidis adherence to intraocular lenses. AB - PURPOSE: To compare the adherence of Staphylococcus epidermidis on AcrySof (Alcon) and poly(methyl methacrylate) (PMMA) intraocular lenses (IOLs) treated with vancomycin, teicoplanin, and cefuroxime. SETTING: Kocaeli University, School of Medicine, Kocaeli, Turkey. METHODS: The lenses were contaminated with S epidermidis solutions containing 10(8) colony-forming units. Intraocular lenses were placed in sterile triptic soy broth after being held in antibiotic solutions for 15 minutes. After that, sonication and vortex procedures were performed to remove all the attached bacteria that could not be removed by antibiotics. Ten microliters from each broth were taken and inoculated into sheep blood agar. The colonies were counted overnight. The statistical analyses were made using Mann Whitney U and Kruskal-Wallis tests, and a P value less than 0.05 was considered statistically significant. RESULTS: Regarding the inhibitory effect of antibiotics on bacterial adhesion, there was no statistically significant difference between AcrySof and PMMA lenses. In the cefuroxime group, the mean numbers of colony-forming units on AcrySof and PMMA lenses were 35.2 +/- 6.94 and 30.8 +/- 18.69, respectively (P>.05). In the teicoplanin group, the mean number of colony-forming units on AcrySof lenses was 100.1 +/- 20.97 and 70.6 +/- 43.75 on the PMMA lenses. The adherence in the vancomycin group was 245 +/- 273.74 colony-forming units for AcrySof lenses and 159.8 +/- 101.94 for PMMA lenses (P>.05). Regarding the inhibitory effect of antibiotics, cefuroxime was the most effective, followed by teicoplanin and vancomycin, respectively. Overall, the mean numbers of colony-forming units on the lenses that were held in cefuroxime, teicoplanin, and vancomycin solutions were 33.0 +/- 13.90, 85.35 +/- 36.66, and 202.4 +/- 205.74, respectively (P = .000). CONCLUSIONS: The results suggest that cefuroxime, teicoplanin, and vancomycin significantly inhibit bacterial adherence to IOLs. The effect of cefuroxime on adherence inhibition was significantly higher than that of teicoplanin and vancomycin. Bacterial adherence is an important factor in bacterial virulence. Antibiotics, especially cefuroxime, can successfully inhibit bacterial adherence. PMID- 16246791 TI - Pillow case. AB - An 80-year-old man with severe kyphosis and visually significant cataracts in both eyes came to us for care. He had been scheduled for cataract surgery elsewhere, but his surgery was cancelled in the operating room when he could not be positioned adequately beneath the operating microscope. As he sat in a chair in our examining room, he was only able to elevate his head to a position 30 degrees below the horizontal. His cataract surgery was performed safely and effectively by padding the operating table with 14 pillows. Eleven pillows were placed under his buttocks and legs and 3 pillows beneath his head and neck. The operating table was also positioned in maximum reverse Trendelenburg to obtain a good red reflex. PMID- 16246790 TI - Elevated intraocular pressure in secondary piggyback intraocular lens implantation. AB - We report 2 cases of postoperative intraocular pressure (IOP) elevation in secondary piggyback intraocular lens (IOL) implantation without history of glaucoma or ocular hypertension. A 74-year-old woman with myopic pseudophakia and a 68-year-old man with hyperopic pseudophakia received secondary piggyback AcrySof IOL implantation in their left eyes. In both patients, the left IOP gradually increased and sustained around 30 mm Hg for about 1 year. In the first, IOP continued elevating despite topical and systemic medications. There was an episode of pupillary block in the second. Gonioscopically, heavier trabecular meshwork pigmentation in their left eyes was observed. Because of this, the 2 IOLs implanted were removed and replaced by an adequate IOL and trabeculotomy was performed in the former. The AcrySof IOL has a truncated optic edge, which increases the risk for chafing the iris, resulting in pigment dispersion syndrome; thus, it would be a poor choice for a sulcus-placed piggyback implantation. PMID- 16246792 TI - Cocooning of an iris-fixated intraocular lens in a 3-year-old child after perforating injury: clinicopathologic correlation. AB - We present a 3-year-old patient who developed extensive noninflammatory fibrous membrane formation in the anterior chamber involving the present iris-fixated intraocular lens (IOL), with subsequent need for explantation. The child had had repair of a corneal injury, crystalline lens extraction and Artisan iris-claw IOL (Ophthec) implantation after a penetrating injury 2 months before. Histopathologic analyses were consistent with a retrocorneal membrane that originated from the corneal stroma and formed a "cocoon" membrane on the iris claw IOL. PMID- 16246793 TI - Spontaneous recovery in phacolytic glaucoma. AB - We report a case of phacolytic glaucoma in which spontaneous absorption of the hypermature lens allowed a patient who refused surgery to recover a normal pressure and satisfactory visual acuity. PMID- 16246794 TI - Pupillary capture of implantable contact lens after blunt trauma. AB - A 33-year-old woman visited our clinic with blurry vision and periorbital swelling after experiencing blunt trauma to left eye. Ten months earlier, she had implantable contact lens (ICL) implantation in the left eye. Biomicroscopic examination showed that that 1 footplate of the ICL was entrapped in the pupillary aperture at the 7 o'clock position and the ICL was placed vertically. The patient had limited ocular movement in lateral gaze, and the computed tomography showed a medial orbital wall fracture. Pupillary capture of the ICL was surgically corrected with an iris manipulator under topical anesthesia. After the ICL was repositioned, the patient's uncorrected visual acuity was restored to 20/32, as before the injury. Pupillary capture of the ICL may occur after blunt ocular trauma. PMID- 16246795 TI - Endophthalmitis after microincision cataract surgery. AB - We report the first case of streptococcal endophthalmitis after uneventful right bimanual phacoemulsification. Microincision cataract surgery is perceived to be minimally invasive as smaller wounds are equated to shortened healing time, increased safety, and reduced risk for postoperative endophthalmitis. Recommendations for modifications in wound construction are discussed. PMID- 16246796 TI - Postoperative localized opacification of the new MemoryLens design: analyses of an explant. AB - We describe a different form of opacification of the new MemoryLens intraocular lens model CV232, leading to a decrease in visual acuity and requiring explantation. A well-circumscribed, centrally/paracentrally located opacification of the optic was observed, with the aspect of a small "lens inside the lens." Histochemical and surface analyses revealed the presence of calcified deposits within a void in the optic substance. This void was seen as a linear optic breach in analysis of an optic cylindrical block cut through the area of opacification, from a sagittal view. The origins of the optic void are unclear. PMID- 16246797 TI - Schisis/cavitation of the MemoryLens optic. PMID- 16246798 TI - Additional uses of trypan blue staining during cataract surgery. PMID- 16246799 TI - Subretinal dislocation of the crystalline lens: unusual complication of phacoemulsification. PMID- 16246800 TI - Late capsular block syndrome after can-opener capsulotomy. PMID- 16246801 TI - Early refractive stabilization after temporal phacoemulsification: what is the optimum time for spectacle prescription? PMID- 16246802 TI - Comparison of dexmedetomidine and midazolam sedation for cataract surgery under topical anesthesia. PMID- 16246804 TI - Hurricane Katrina. PMID- 16246803 TI - Phacoemulsification in patients with thrombocytopenia. PMID- 16246805 TI - Thoughts on Katrina. PMID- 16246807 TI - Cute? ... or "Cutesy"? PMID- 16246808 TI - Nursing activities and use of time in the postanesthesia care unit. AB - Specialty areas, such as the PACU, are currently characterized by a shortage of qualified nursing personnel along with the pressure for a most cost-conscious function of services. These, in combination with both the increasing acuity of patients and the advances of science and technology, have rendered necessary the investigation of how existing personnel can be better used. The work sampling approach has been widely used in different clinical settings to offer data about the amount of time nurses devote to specific activities. The aim of the current study was to categorize and quantify the activities of nurses employed in the PACU of the General University Hospital of Patras, Greece, and to identify differences with regard to shifts, varying nurse-patient ratios, and nurses' experiences. In the first phase, a classification form, which included all nursing activities, was constructed. This was based on literature review, researchers' experience, and semistructured personnel interviews. In the second phase, the researchers observed all PACU nurses, and activities were recorded on an hourly check sheet. A total of 4,320 observations were collected from 9/1/2004 to 9/30/2004. Nurses spent 35.2% of their time on direct clinical care, 11.6% on patient assessment, 7.0% on communication with patients, 7.2% on communication with other persons, 8.6% on clerical nursing duties, 9.3% on documentation, 2.3% on non-nursing duties, and 18.8% on personal activities. Variations in the use of time were found between shifts, different nurse-patient ratios, and PACU experience. Methods of decreasing time when the nurse-patient ratio is inappropriate and increasing time allocated to direct care activities are proposed. PMID- 16246809 TI - Sleep/sedation in children undergoing EEG testing: a comparison of chloral hydrate and music therapy. AB - This study included a total of 60 pediatric patients ranging from 1 month through 5 years of age. The effects of chloral hydrate and music therapy were evaluated and compared as means of safe and effective ways to achieve sleep/sedation in infants and toddlers undergoing EEG testing. The results of the study indicate that music therapy may be a cost-effective, risk-free alternative to pharmacological sedation. PMID- 16246810 TI - Effects of preoperative teaching of the use of a pain scale with patients in the PACU. AB - Clinical observations have shown that patients arriving in the PACU who had no experience using a pain scale had more difficulty rating their pain in the immediate postoperative period. The purpose of this pilot study was to determine if preoperative instruction on the use of a pain scale would improve the patient's ability to self-report pain in the Phase I PACU. The sample consisted of 50 English speaking, orthopedic patients between the ages of 19 and 75 years. A visual numerical rating scale (NRS) for pain was used to teach patients in the holding area. Twenty-six of these patients had previous experience with the NRS and 24 had no experience. Of those with experience, 21 (80%) could use the pain scale in the PACU. Of the 24 patients who were taught in the holding area, 20 (85%) could use the pain scale to rate their pain in the PACU. The results of this study suggest that if patients have previous experience with a pain scale, or if they are taught preoperatively, they can more effectively self-report pain postoperatively. Future studies should be conducted to involve a larger sample, a variety of surgeries, and non-English-speaking patients. PMID- 16246811 TI - Implementing the 2005 National Patient Safety Goals in the ambulatory surgery setting. PMID- 16246812 TI - Extended-release epidural morphine (DepoDur). PMID- 16246813 TI - Outcomes research: a paradigm shift for nursing research. PMID- 16246814 TI - The stressed patient with diabetes mellitus. PMID- 16246815 TI - Who is vulnerable? PMID- 16246816 TI - Pain-related fear at the start of a new low back pain episode. AB - Previous research supports the fear-avoidance model in explaining chronic low back pain (LBP) disability. The aims of the present study were to determine: (1) whether fear-avoidance model variables are associated already during acute stages of LBP and (2) whether (increases in) pain-related fear are associated with other patient characteristics routinely assessed by the General Practitioner (GP). General practice patients consulting because of a new episode of LBP completed questionnaires on pain-related fear, avoidance, pain and disability. A sample of 247 acute LBP patients (median duration of current episode was 5 days) was collected. Significant associations were found between pain intensity, pain related fear, avoidance behaviour and disability, but correlations were generally modest. A strong association was found between pain and disability. Pain-related fear was slightly higher in patients reporting low job satisfaction and in those taking bedrest. These results suggest that the fear-avoidance model as it was developed and tested in chronic LBP, might not entirely apply to acute LBP patients. Future research should focus on the transition from acute to chronic LBP and the shifts that take place between fear-avoidance model associations. PMID- 16246817 TI - Sexual dysfunction and chronic pain: the role of psychological variables and impact on quality of life. AB - We report two studies examining the prevalence of sexual dysfunction, and the role of psychological variables, including quality of life, on sexual activity in patients at the commencement of an outpatient cognitive-behavioural pain management programme. In Study 1, 151 patients with non-cancer pain, predominantly of musculoskeletal origin, completed a range of standardised measures, including the Pain Disability Index, Beck Depression Inventory and Coping Strategies Questionnaire. Sexual dysfunction was common, and using stepwise multiple regression analysis was found to be more frequently reported by those with greater disability and depression, shorter pain duration, and infrequent use of coping self-statements. Study 2 was a pilot investigation of the impact of sexual dysfunction on quality of life (as measured by the WHOQOL 100) in a similar sample (n=41). Although sexual dysfunction was again commonly reported, subjects perceived it had less importance in quality of life than did other factors. The combined results support the previously proposed notion of adaptation to the impact of chronic illness on sexual function. In conclusion, sexual dysfunction is common in this population and is predicted by psychological factors and pain duration. However, other issues impact more significantly on quality of life. Therapeutic approaches to sexual dysfunction in these patients might best be focused on improving psychological factors, particularly depression and coping skills. PMID- 16246818 TI - Delayed onset muscle soreness in neck/shoulder muscles. AB - The aim of the present study is to: (1) induce delayed onset muscle soreness (DOMS) in the neck and shoulder muscles; (2) compare the pressure pain sensitivity of muscle belly with that of musculotendinous tissue after DOMS; (3) examine the gender differences in the development of DOMS. An eccentric shoulder exercise was developed to induce DOMS on neck/shoulder muscles using a specially designed dynamometer. Eccentric shoulder contraction consisted of 5 bouts, each bout lasted 3min, with 3min rest period between each bout. The right shoulder was elevating against a downward pressure force of 110% maximal voluntary contraction force exerted by the dynamometer. Pressure pain thresholds (PPT) of 11 sites (seven sites measured were muscle belly and four sites were myotendinous area) on neck/shoulder region were measured before, immediately after, 24 and 48h after exercise. Pain intensity, pain area and index of McGill pain questionnaire were assessed and all were increased after exercise. DOMS was induced in the shoulder muscles. PPT was significantly decreased and reached lowest values at 24h. The muscle belly sites are more sensitive to pain than the musculotendinous sites. No gender differences were found in any of the parameters used to assess the development of DOMS. DOMS did not distribute evenly in the neck/shoulder region. Soreness after exercise in the neck and shoulder seems not to be among the conditions that produce predominant musculoskeletal pain in females. PMID- 16246820 TI - Effect of long-term neck muscle training on pressure pain threshold: a randomized controlled trial. AB - Muscle tenderness has been measured in several studies to evaluate effectiveness of treatment methods, but only short-term results have been reported so far. The aim of the present study was to evaluate the long-term effects of two different muscle training methods on the pressure pain threshold of neck muscles in women with neck pain. Altogether 180 woman with chronic, non-specific neck pain were randomized into three groups: neck muscle endurance training, neck muscle strength training and control groups. The main outcome measures included pressure pain threshold measurement at six muscle sites and on the sternum. Neck pain was assessed by a visual analogue scale (VAS). At the 12-month follow-up statistically significantly higher pressure pain threshold values were obtained in both training groups at all muscle sites compared to the baseline, while no significant change occurred in the controls. Significantly higher changes in pressure pain threshold were detected at all six sites in the strength training group and at four out of six sites in the endurance training group compared to the control group. This is the first study to show an increase in pressure pain thresholds as a result of long-term muscle training. A decrease in neck pain was associated with reduced pressure pain sensitivity in neck muscles, showing that the pressure pain threshold may be a useful outcome measure of the effectiveness of neck muscle rehabilitation. PMID- 16246819 TI - Are migraineurs hypersensitive? A test of the stimulus processing disorder hypothesis. AB - The concept of hypersensitivity in migraineurs was advanced mainly on the basis of studies on information processing in which increased amplitudes and reduced habituation in cortical evoked and event related potentials were found in migraine sufferers. The present investigation examined whether migraineurs exhibit hypersensitivity within three different experimental paradigms and various non electrocortical response parameters. Samples of 24 migraine, 19 tension-type headache sufferers, and 24 normal controls were compared regarding their subjective estimation of intensity and discomfort due to visual and acoustical stimuli. Subjects also participated in an experiment using the eyeblink startle response paradigm. In a last experimental task the Stroop test was applied. The trait variables emotionality, arousal, and extraversion were also measured. None of the experimental tasks revealed the predicted hypersensitivity of migraineurs in relation to the control samples. The series of experiments was conducted a second time with half of the participants in order to replicate the findings. The conclusions remained the same. The results of earlier studies on cortical processing can not be interpreted as demonstrating general hypersensitivity in the sense of a dispositional trait in migraine afflicted individuals irrespective of the involved response system. PMID- 16246821 TI - Psychosocial functioning of mothers with chronic pain: a comparison to pain-free controls. AB - This study compared the functioning of mothers experiencing chronic pain and control mothers on a range of psychosocial variables. Participants included 39 mothers with chronic pain conditions ranging from migraine and arthritis to chronic neck and back pain and 35 control mothers with out chronic pain. Analyses indicated that mothers with chronic pain experienced more physical, psychological and social difficulties when compared to controls. More difficulties were reported in completing day-to-day parenting tasks in mothers with chronic pain. Consistent with the biopsychosocial model of chronic pain, psychosocial variables accounted for approximately half of the variance in chronic pain mothers' physical functioning scores. The importance of psychological variables in the experience of chronic pain, the potential reduction in parenting efficacy and the risk that these influences hold for children are discussed. PMID- 16246822 TI - Methods of translating NMR proton distances into their corresponding heavy atom distances for protein structure prediction with limited experimental data. AB - This paper proposes a strategy to translate experimental 1H NMR proton distance restraints into their corresponding heavy atom distance restraints for the purpose of protein structure prediction. The relationships between interproton distances and the corresponding heavy atom distances are determined by studying well-resolved X-ray protein structures. The data from the interproton distances of amide protons, alpha-protons, beta-protons and side chain methyl protons are plotted against the corresponding heavy atoms in scatter plots and then fitted with linear equations for lower bounds, upper bounds and optimal fits. We also transform the scatter plots into two-dimensional heat maps and three-dimensional histograms, which identify the regions where data points concentrate. The common interproton distances between amide protons, alpha-protons, beta-protons in alpha helices, anti-parallel beta-sheets and parallel beta-sheets are also tabulated. We have found several patterns emerging from the distance relationships between heavy atom pairs and their corresponding proton pairs. All our upper bound, lower bound and optimal fit results for translating the interproton distance into their corresponding heavy atom distances are tabulated. PMID- 16246823 TI - Enhancing the stability of microsomal cytochrome b5: a rational approach informed by comparative studies with the outer mitochondrial membrane isoform. AB - The outer mitochondrial membrane isoform of mammalian cytochrome b5 (OM b5) is much less prone to lose heme than the microsomal isoform (Mc b5), with a conserved difference at position 71 (leucine versus serine) playing a major role. We replaced Ser71 in Mc b5 with Leu, with the prediction that it would retard heme loss by diminishing polypeptide expansion accompanying rupture of the histidine to iron bonds. The strategy was partially successful in that it slowed dissociation of heme from its less stable orientation in bMc b5 (B). Heme dissociation from orientation A was accelerated to a similar extent, however, apparently owing to increased binding pocket dynamic mobility related to steric strain. A second mutation (L32I) guided by results of previous comparative studies of Mc and OM b5s diminished the steric strain, but much greater relief was achieved by replacing heme with iron deuteroporphyrin IX (FeDPIX). Indeed, the stability of the Mc(S71L) b5 FeDPIX complex is similar to that of the FeDPIX complex of OM b5. The results suggest that maximizing heme binding pocket compactness in the apo state is a useful general strategy for increasing the stability of engineered or designed proteins. PMID- 16246824 TI - An evolution-based analysis scheme to identify CO2/O2 specificity-determining factors for ribulose 1,5-bisphosphate carboxylase/oxygenase. AB - Ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBisCo) catalyzes a rate limiting step in photosynthetic carbon assimilation (reacting with CO2) and its competitive photo-respiratory carbon oxidation (reacting with O2). RuBisCo enzyme with an enhanced CO2/O2 specificity would boost the ability to make great progress in agricultural production and environmental management. RuBisCos in marine non-green algae, resulting from an earlier endo-symbiotic event, diverge greatly from those in green plants and cyanobacteria and, further, have the highest CO2/O2 specificity whereas RuBisCos in cyanobacteria have the lowest. We assumed that there exist different levels of CO2/O2 specificity-determining factors, corresponding to different evolutionary events and specificity levels. Based on this assumption, we devised a scheme to identify these substrate determining factors. From this analysis, we are able to discover different categories of the CO2/O2 specificity-determining factors that show which residue substitutions account for (relatively) small specificity changes, as happened in green plants, or a tremendous enhancement, as observed in marine non-green algae. Therefore, the analysis can improve our understanding of molecular mechanisms in the substrate specificity development and prioritize candidate specificity determining surface residues for site-directed mutagenesis. PMID- 16246825 TI - Left ventricular hypertrophy, apoptosis, and progression to heart failure in severe aortic stenosis. PMID- 16246826 TI - Social inhibition modulates the effect of negative emotions on cardiac prognosis following percutaneous coronary intervention in the drug-eluting stent era. AB - AIMS: Negative emotions have an adverse effect on cardiac prognosis. We investigated whether social inhibition (inhibited self-expression in social interaction) modulates the effect of negative emotions on clinical outcome following percutaneous coronary intervention (PCI). METHODS AND RESULTS: Eight hundred and seventy-five consecutive patients from the RESEARCH registry (Erasmus Medical Centre, Rotterdam) completed depression, anxiety, negativity (negative emotions in general), and social inhibition scales 6 months following PCI. The endpoint was major adverse cardiac event (MACE-death, myocardial infarction, coronary artery bypass graft (CABG), or PCI) at 9 months following assessment. There were 100 MACE; patients who were high in both negativity and inhibition were at increased risk of MACE (38/254=15%) when compared with high negativity/low inhibition patients (13/136=10%; P=0.018). Depression (P=0.23) or anxiety (P=0.63) did not explain away this moderating effect of inhibition. High negativity/high inhibition (HR=1.92, 95%CI 1.22-3.01, P=0.005) and previous CABG (HR=1.90, 95%CI 1.04-3.47, P=0.038) were independent predictors of MACE. Patients with high negativity but low inhibition were not at increased risk (P=0.76). High negativity/high inhibition also independently predicted death/MI (n=20) as a more specific endpoint (HR=5.85, P=0.001). CONCLUSION: The interaction effect of social inhibition and negative emotions, rather than negative emotions per se, predicted poor clinical outcome following PCI. Social inhibition should not be overlooked as a modulating factor. PMID- 16246827 TI - Coronary artery disease, heart failure, and cardiac natriuretic peptides in the middle. PMID- 16246828 TI - Cardiac lymphoma. PMID- 16246829 TI - Ventricular tachycardia and cardiac sarcoidosis: correspondence between MRI and electrophysiology. PMID- 16246830 TI - Sirolimus-eluting stent fracture with thrombus, visualization by optical coherence tomography. PMID- 16246831 TI - Repair of all the components of the syndrome of aortic regurgitation and VSD. PMID- 16246832 TI - Transthoracic echocardiography of Hodgkin lymphoma in the upper anterior mediastinum causing compression of the great vessels. PMID- 16246833 TI - Hotline sessions of the 27th European congress of cardiology. PMID- 16246834 TI - Magnetic resonance imaging after percutaneous closure of a ventricular septal defect complicating myocardial infarction. PMID- 16246835 TI - Congenital left atrial appendage aneurysm in a 2-year-old boy. PMID- 16246836 TI - Left ventricular pseudoaneurysm. PMID- 16246838 TI - NF-kappaB protects macrophages from lipopolysaccharide-induced cell death: the role of caspase 8 and receptor-interacting protein. AB - Macrophages play a pivotal role in the pathogenesis of a variety of diseases. These studies were performed to characterize the mechanisms by which Toll-like receptor 4 (TLR4)-mediated NF-kappaB activation promotes resistance to cell death in macrophages. When NF-kappaB activation was inhibited by a super-repressor, IkappaBalpha, the TLR4 ligand lipopolysaccharide induced the activation of caspase 8, the loss of mitochondrial transmembrane potential (DeltaPsim), and apoptotic cell death in macrophages. The inhibition of caspase 8 activation suppressed DNA fragmentation but failed to protect macrophages against the loss of DeltaPsim and resulted in necrotic cell death. In contrast, the reduction of receptor-interacting protein 1 suppressed the loss of DeltaPsim and inhibited apoptotic cell death. Further, when caspase 8 activation was suppressed, the knock down of receptor-interacting protein inhibited the loss of DeltaPsim and necrotic cell death. These observations demonstrate that following TLR4 ligation by lipopolysaccharide, NF-kappaB is a critical determinant of macrophage life or death, whereas caspase 8 determines the pathway employed. PMID- 16246837 TI - Biophysical characterization of the signature domains of thrombospondin-4 and thrombospondin-2. AB - The signature domain of thrombospondins consists of tandem epidermal growth factor-like modules, 13 calcium-binding repeats, and a lectin-like module. Although very similar, the signature domains of thrombospondin-1 and -2 differ in several potentially important ways from the domains of thrombospondin-3, -4, and 5. We have compared matching recombinant segments representing the signature domains of thrombospondin-2 and -4. In the presence of 2 mM CaCl2, the far UV circular dichroism spectra of thrombospondin-2 and -4 constructs contain a strong negative band at 202 nm, but only the thrombospondin-2 construct has a band at 216 nm. Chelation of calcium shifted the negative bands to lower magnitudes. Titrations of the spectra demonstrated lower cooperativity and affinity for binding of calcium to thrombospondin-4 compared with thrombospondin-2. Atomic absorption spectroscopy demonstrated that the thrombospondin-4 constructs bind seven less calcium than the thrombospondin-2 construct at 0.6 mM CaCl2. In 2 mM CaCl2, the near UV circular dichroism spectra of thrombospondin-2, but not thrombospondin-4, contain a positive band at 292 nm that disappears upon calcium chelation. Intrinsic fluorescence spectra for both proteins were also sensitive to calcium, but the changes were simpler and more marked for thrombospondin-2 than for thrombospondin-4. In differential scanning calorimetry, the thrombospondin-2 construct melted in two distinct transitions at 53.5 and 81.8 degrees C, whereas the first transition for thrombospondin-4 constructs was observed at 63.5 degrees C. Thus, the studies revealed significant differences between the signature domains of thrombospondin-2 and thrombospondin-4 in calcium binding, fine structure, and inter-modular interactions. PMID- 16246840 TI - Nitric oxide negatively regulates Fas CD95-induced apoptosis through inhibition of ubiquitin-proteasome-mediated degradation of FLICE inhibitory protein. AB - Stimulation of cell surface Fas (CD95) results in recruitment of cytoplasmic proteins and activation of caspase-8, which in turn activates downstream effector caspases leading to programmed cell death. Nitric oxide (NO) plays a key role in the regulation of apoptosis, but its role in Fas-induced cell death and the underlying mechanism are largely unknown. Here we show that stimulation of the Fas receptor by its ligand (FasL) results in rapid generation of NO and concomitant decrease in cellular FLICE inhibitory protein (FLIP) expression without significant effect on Fas and Fas-associated death domain (FADD) adapter protein levels. FLIP down-regulation as well as caspase-8 activation and apoptosis induced by FasL were all inhibited by the NO-liberating agent sodium nitroprusside and dipropylenetriamine NONOate, whereas the NO synthase inhibitor aminoguanidine and NO scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethyl imidazoline-1-oxyl-3-oxide (PTIO) had opposite effects, indicating an anti apoptotic role of NO in the Fas signaling process. FasL-induced down-regulation of FLIP is mediated by a ubiquitin-proteasome pathway that is negatively regulated by NO. S-nitrosylation of FLIP is an important mechanism rendering FLIP resistant to ubiquitination and proteasomal degradation by FasL. Deletion analysis shows that the caspase-like domain of FLIP is a key target for S nitrosylation by NO, and mutations of its cysteine 254 and cysteine 259 residues completely inhibit S-nitrosylation, leading to increased ubiquitination and proteasomal degradation of FLIP. These findings indicate a novel pathway for NO regulation of FLIP that provides a key mechanism for apoptosis regulation and a potential new target for intervention in death receptor-associated diseases. PMID- 16246839 TI - Characterization of an ERK-binding domain in microphthalmia-associated transcription factor and differential inhibition of ERK2-mediated substrate phosphorylation. AB - Efficient and specific signaling by mitogen-activated protein kinases (MAPKs) is enhanced by docking sites found on many MAPK substrates and regulators. Here we show that the MAPKs ERK1 and ERK2 form a stable complex (Kd approximately 6 microm) with their substrate the microphthalmia-associated transcription factor (MITF). Complex formation requires a domain of MITF of approximately 100 residues that is nearby, but C-terminal to, the MAPK phosphorylation site at Ser73. MITF derivatives lacking this ERK-binding domain do not bind ERK2 and are phosphorylated less efficiently by ERK2. The ERK-binding domain of MITF bears no obvious resemblance to previously characterized MAPK docking motifs; in particular, it does not contain a consensus D-site. Consistent with this, ERK2 MITF binding does not require the integrity of the CD/sevenmaker region of ERK2. Furthermore, D-site peptides, which are able to potently inhibit ERK2-mediated phosphorylation of the Elk-1 transcription factor (IC50= 3 microm), are relatively poor inhibitors of ERK2-mediated phosphorylation of MITF, exhibiting >15-fold selectivity for inhibition of Elk-1 versus MITF. These observations demonstrate substrate-selective kinase inhibition: the possibility that small molecules that target docking interactions may be used to selectively inhibit the phosphorylation of a subset of the substrates of a kinase. PMID- 16246841 TI - Shigella Spa33 is an essential C-ring component of type III secretion machinery. AB - Type III secretion machinery (TTSM), composed of a needle, a basal body, and a C ring compartment, delivers a subset of effectors into host cells. Here, we show that Shigella Spa33 is an essential component of the C-ring compartment involved in mediating the transit of various TTSM-associated translocated proteins. Electron microscopic analysis and pull-down assay revealed Spa33 to be localized beneath the TTSM via interaction with MxiG and MxiJ (basal body components). Spa33 is also capable of interacting with Spa47 (TTSM ATPase), MxiK, MxiN (required for the transit of MxiH, the needle component), Spa32 (required for determining needle length), and several effectors. Genetic and functional analyses of the Spa33 C-terminal region, which is highly conserved in the SpaO YscQ-HrcQ(B)-FliN family, indicate that some of the conserved residues are crucial for needle formation via interactions with MxiN. Thus, Spa33 plays a central role as the C-ring component in recruiting/exporting TTSM-associated proteins. PMID- 16246842 TI - FlhB regulates ordered export of flagellar components via autocleavage mechanism. AB - The bacterial flagellum is a predominantly cell-external super-macromolecular construction whose structural components are exported by a flagellum-specific export apparatus. One of the export apparatus proteins, FlhB, regulates the substrate specificity of the entire apparatus; i.e. it has a role in the ordered export of the two main groups of flagellar structural proteins such that the cell proximal components (rod-/hook-type proteins) are exported before the cell-distal components (filament-type proteins). The controlled switch between these two export states is believed to be mediated by conformational changes in the structure of the C-terminal cytoplasmic domain of FlhB (FlhB(C)), which is consistently and specifically cleaved into two subdomains (FlhB(CN) and FlhB(CC)) that remain tightly associated with each other. The cleavage event has been shown to be physiologically significant for the switch. In this study, the mechanism of FlhB cleavage has been more directly analyzed. We demonstrate that cleavage occurs in a heterologous host, Saccharomyces cerevisiae, deficient in vacuolar proteinases A and B. In addition, we find that cleavage of a slow-cleaving variant, FlhB(C)(P270A), is stimulated in vitro at alkaline pH. We also show by analytical gel-filtration chromatography and analytical ultracentrifugation experiments that both FlhB(C) and FlhB(C)(P270A) are monomeric in solution, and therefore self-proteolysis is unlikely. Finally, we provide evidence via peptide analysis and FlhB cleavage variants that the tertiary structure of FlhB plays a significant role in cleavage. Based on these results, we propose that FlhB cleavage is an autocatalytic process. PMID- 16246843 TI - Ferrous ions and reactive oxygen species increase antigen-binding and anti inflammatory activities of immunoglobulin G. AB - Polyspecific antibodies represent a first line of defense against infection and regulate inflammation, properties hypothesized to rely on their ability to interact with multiple antigens. We demonstrated that IgG exposure to pro oxidative ferrous ions or to reactive oxygen species enhances paratope flexibility and hydrophobicity, leading to expansion of the spectrum of recognized antigens, regulation of cell proliferation, and protection in experimental sepsis. We propose that ferrous ions, released from transferrin and ferritin at sites of inflammation, synergize with reactive oxygen species to modify the immunoglobulins present in the surrounding microenvironment, thus quenching pro-inflammatory signals, while facilitating neutralization of pathogens. PMID- 16246844 TI - Inducible expression of Tau repeat domain in cell models of tauopathy: aggregation is toxic to cells but can be reversed by inhibitor drugs. AB - We generated several cell models of tauopathy in order to study the mechanisms of neurodegeneration in diseases involving abnormal changes of tau protein. N2a neuroblastoma cell lines were created that inducibly express different variants of the repeat domain of tau (tau(RD)) when exposed to doxycycline (Tet-On system). The following three constructs were chosen: (i) the repeat domain of tau that coincides with the core of Alzheimer paired helical filaments; (ii) the repeat domain with the deletion mutation DeltaK280 known from frontotemporal dementia and highly prone to spontaneous aggregation; and (iii) the repeat domain with DeltaK280 and two proline point mutations that inhibit aggregation. The comparison of wild-type, pro-aggregation, and anti-aggregation mutants shows the following. (a) Aggregation of tau(RD) is toxic to cells. (b) The degree of aggregation and toxicity depends on the propensity for beta-structure. (c) Soluble mutants of tau(RD) that cannot aggregate are not toxic. (d) Aggregation is preceded by fragmentation. (e) Fragmentation of tau(RD) in cells is initially due to a thrombin-like protease activity. (f) Phosphorylation of tau(RD) (at KXGS motifs) precedes aggregation but is not correlated with the degree of aggregation. (g) Aggregates of tau(RD) disappear when the expression is silenced, showing that aggregation is reversible. (h) Aggregation can be prevented by drugs and even pre-formed aggregates can be dissolved again by drugs. Thus, the cell models open up new insights into the relationship between the structure, expression, phosphorylation, aggregation, and toxicity of tau(RD) that can be used to test current hypotheses on tauopathy and to develop drugs that prevent the aggregation and degeneration of cells. PMID- 16246845 TI - Early events in the fibrillation of monomeric insulin. AB - Insulin has a largely alpha-helical structure and exists as a mixture of hexameric, dimeric, and monomeric states in solution, depending on the conditions: the protein is monomeric in 20% acetic acid. Insulin forms amyloid like fibrils under a variety of conditions, especially at low pH. In this study we investigated the fibrillation of monomeric human insulin by monitoring changes in CD, attenuated total reflectance-Fourier transform infrared spectroscopy, 8 anilinonaphthalenesulfonic acid fluorescence, thioflavin T fluorescence, dynamic light scattering, and H/D exchange during the initial stages of the fibrillation process to provide insight into early events involving the monomer. The results demonstrate the existence of structural changes occurring before the onset of fibril formation, which are detectable by multiple probes. The data indicate at least two major populations of oligomeric intermediates between the native monomer and fibrils. Both have significantly non-native conformations, and indicate that fibrillation occurs from a beta-rich structure significantly distinct from the native fold. PMID- 16246846 TI - Characterization of a defensin from the oyster Crassostrea gigas. Recombinant production, folding, solution structure, antimicrobial activities, and gene expression. AB - In invertebrates, defensins were found in arthropods and in the mussels. Here, we report for the first time the identification and characterization of a defensin (Cg-Def) from an oyster. Cg-def mRNA was isolated from Crassostrea gigas mantle using an expressed sequence tag approach. To gain insight into potential roles of Cg-Def in oyster immunity, we produced the recombinant peptide in Escherichia coli, characterized its antimicrobial activities, determined its solution structure by NMR spectroscopy, and quantified its gene expression in vivo following bacterial challenge of oysters. Recombinant Cg-Def was active in vitro against Gram-positive bacteria but showed no or limited activities against Gram negative bacteria and fungi. The activity of Cg-Def was retained in vitro at a salt concentration similar to that of seawater. The Cg-Def structure shares the so-called cystine-stabilized alpha-beta motif (CS-alphabeta) with arthropod defensins but is characterized by the presence of an additional disulfide bond, as previously observed in the mussel defensin (MGD-1). Nevertheless, despite a similar global fold, the Cg-Def and MGD-1 structures mainly differ by the size of their loops and by the presence of two aspartic residues in Cg-Def. Distribution of Cg-def mRNA in various oyster tissues revealed that Cg-def is mainly expressed in mantle edge where it was detected by mass spectrometry analyses. Furthermore, we observed that the Cg-def messenger concentration was unchanged after bacterial challenge. Our results suggest that Cg-def gene is continuously expressed in the mantle and would play a key role in oyster by providing a first line of defense against pathogen colonization. PMID- 16246847 TI - Why is the Arg5-Glu13 salt bridge conserved in mammalian alpha-defensins? AB - Mammalian alpha-defensins, expressed primarily in leukocytes and epithelia, kill a broad range of microbes, constituting one of the first lines of innate immune defense against infection. Nine amino acid residues, including six cysteines, one glycine, and a pair of oppositely charged residues Arg/Glu, are conserved in the otherwise diverse sequences of all known mammalian alpha-defensins. Structural analysis indicates that the two charged residues form a salt bridge, likely stabilizing a protruding loop in the molecule. To investigate the structural and functional roles of the conserved Arg5-Glu13 salt bridge in alpha-defensins, we chemically prepared human neutrophil alpha-defensin 2 (HNP2) and five HNP2 analogs, R5E/E13R, E13Q, E13R, R5T/E13Y, and R14A. In contrast to HNP2 and R14A HNP2, none of the four salt bridge analogs was capable of folding into a native conformation in the context of isolated defensin domains. However, when covalently attached to the 45-residue pro-HNP2 propeptide, the salt bridge analogs of HNP2 in their pro-forms all folded productively, suggesting that the Arg5-Glu13 salt bridge is not required for correct pro-alpha-defensin folding. When assayed against both Escherichia coli and Staphylococcus aureus, the six alpha-defensins showed bactericidal activity that correlated with the number of net positive charges carried by individual molecules in the panel, irrespective of whether or not the Arg5-Glu13 salt bridge was decimated, suggesting that Arg5 and Glu13 are not functionally conserved. Proteolytic resistance analysis with human neutrophil elastase, one major protease contained in azurophils with HNPs, revealed that destabilization of the salt bridge dramatically accelerated defensin degradation by the enzyme. Thus, we propose that the Arg5-Glu13 salt bridge found in most mammalian alpha-defensins is conserved for defensin in vivo stability. PMID- 16246848 TI - Proliferation of pulmonary interstitial fibroblasts is mediated by transforming growth factor-beta1-induced release of extracellular fibroblast growth factor-2 and phosphorylation of p38 MAPK and JNK. AB - Idiopathic pulmonary fibrosis (IPF; a progressive lung disease) is characterized by parenchymal remodeling with enlarged air spaces called honeycomb cysts and palisades of fibroblasts called fibroblast foci. In IPF, lung epithelial cells covering honeycomb cysts and fibroblast foci aberrantly express the active conformation of the potent fibrogenic cytokine transforming growth factor-beta1 (TGF-beta1). Using explanted rat lung slices, we transfected alveolar epithelial cells with the retrovirus pMX containing a site-directed mutation in which Cys223 and Cys225 were substituted with serines, resulting in release of biologically active TGF-beta1 and fibroblast proliferation and remodeling that resembled IPF. Fibroblasts obtained from transfected explants and in culture for 6 weeks incorporated 6.59 +/- 1.55-fold more [3H]thymidine compared with control fibroblasts without transfection or fibroblasts obtained from transfected explants cultured with antibody to fibroblast growth factor-2 (FGF-2). Primary lung fibroblasts obtained from normal rat lungs cultured with TGF-beta1 expressed increased levels of phosphorylated p38 MAPK and JNK, but not ERK1/2. The presence of TGF-beta1 caused an immediate release of extracellular FGF-2 from primary pulmonary fibroblasts; and in the presence of anti-FGF-2 antibody, phosphorylated p38 MAPK and JNK were abrogated. TGF-beta inhibits cell proliferation by suppression of c-Myc and induction of p15INK46, p21CIP1, or p27KIP. Fibroblasts cultured with TGF-beta1 showed no regulation of c-Myc or induction of p15INK46, p21CIP1,or p27KIP. These findings suggest that pulmonary fibroblasts may not respond to the anti-proliferative effects of TGF-beta1, but proliferate in response to TGF-beta1 indirectly by the release of FGF-2, which induces phosphorylation of p38 MAPK and JNK. PMID- 16246849 TI - Genecology of Douglas fir in western Oregon and Washington. AB - BACKGROUND AND AIMS: Genecological knowledge is important for understanding evolutionary processes and for managing genetic resources. Previous studies of coastal Douglas fir (Pseudotsuga menziesii var. menziesii) have been inconclusive with respect to geographical patterns of variation, due in part to limited sample intensity and geographical and climatic representation. This study describes and maps patterns of genetic variation in adaptive traits in coastal Douglas fir in western Oregon and Washington, USA. METHODS: Traits of growth, phenology and partitioning were measured in seedlings of 1338 parents from 1048 locations grown in common gardens. Relations between traits and environments of seed sources were explored using regressions and canonical correlation analysis. Maps of genetic variation as related to the environment were developed using a geographical information system (GIS). KEY RESULTS: Populations differed considerably for adaptive traits, in particular for bud phenology and emergence. Variation in bud set, emergence and growth was strongly related to elevation and cool-season temperatures. Variation in bud-burst and partitioning to stem diameter versus height was related to latitude and summer drought. Seedlings from the east side of the Washington Cascades were considerably smaller, set bud later and burst bud earlier than populations from the west side. CONCLUSIONS: Winter temperatures and frost dates are of overriding importance to the adaptation of Douglas fir to Pacific Northwest environments. Summer drought is of less importance. Maps generated using canonical correlation analysis and GIS allow easy visualization of a complex array of traits as related to a complex array of environments. The composite traits derived from canonical correlation analysis show two different patterns of variation associated with different gradients of cool-season temperatures and summer drought. The difference in growth and phenology between the westside and eastside Washington Cascades is hypothesized to be a consequence of the presence of interior variety (P. menziessii var. glauca) on the eastside. PMID- 16246850 TI - The rear or the front in bag and mask oxygenation of the neonates. PMID- 16246851 TI - Meeting and treating cultural difference in primary care: a qualitative interview study. AB - BACKGROUND: Primary care doctors see patients from diverse cultural backgrounds and communication plays an important role in diagnosis and treatment. Communication problems can arise when patient and doctor do not share the same cultural background. OBJECTIVE: The aim of this study was to examine how consultations with immigrant patients are understood by GPs and how GPs manage these consultations. METHODS: Semi-structured interviews with GPs about their experiences with immigrant patients were recorded on audio-tape, transcribed and analysed using a qualitative thematic analysis methodology. A constructivist approach was taken to analysis and interpretation. RESULTS: Culture is not in focus when GPs meet immigrant patients. The consultation is seen as a meeting between individuals, where cultural difference is just one of many individual factors that influence how well doctor and patient understand each other. However, when mutual understanding is poor and the consultation not successful, cultural differences are central. The GPs try to conduct their consultations with immigrant patients in the same way that they conduct all their consultations. There is no specific focus on culture, instead, GPs tend to avoid addressing even pronounced cultural differences. CONCLUSION: This study indicates that cultural difference is not treated in GPs consultation with immigrant patients. Learning about cultural difference's effect on mutual understanding between doctor and patient could improve GPs cross-cultural communication. Increased awareness of the culture the doctor brings to the consultation could facilitate management of cross-cultural consultations. PMID- 16246852 TI - Characterisation of morbidity in a UK, hospital based, obesity clinic. AB - AIM: To identify clinical features which predict those most at risk of co morbidities within an obesity clinic. METHODS: Children attending an obesity clinic had fasting glucose, insulin, and lipids measured prior to a standard oral glucose tolerance test (OGTT). History and examination established birth weight, family history of type 2 diabetes/obesity, pubertal status, and presence of acanthosis nigricans. Central and total fat mass was estimated by bio-impedance. RESULTS: Of the 126 children evaluated, 10.3% (n = 13) had impaired glucose tolerance (IGT); the majority (n = 11) of these would not have been identified on fasting glucose alone. Those with IGT were more likely to have a parental history of type 2 diabetes (relative risk 3.5). IGT was not associated with acanthosis nigricans. Twenty five per cent (n = 19) of those evaluated (n = 75) had evidence of the "metabolic syndrome" (MS). HDL cholesterol and triglyceride levels were related to insulin sensitivity (HOMA-R); HDL cholesterol was also related to birth weight SDS. We observed a trend for those with MS to have a lower birth weight SDS. The severity of obesity did not influence the likelihood of IGT or MS. CONCLUSIONS: Significant numbers of obese children have associated co morbidities. Analysis of fasting blood glucose samples alone is not satisfactory to adequately evaluate glucose homoeostasis. The overall level of obesity does not predict co-morbidities. Special attention should be given to those with parental diabetes and a history of low birth weight who are more likely to have IGT and abnormal lipid profiles respectively. PMID- 16246854 TI - The relationship between parental perceptions of diabetes and glycaemic control. AB - AIM: To measure the relationship between perceived child competence, parental self-efficacy, and children's glycaemic control. METHODS: Cross-sectional outpatient based questionnaire survey of 78 parents of children aged 6-12 years with insulin dependent diabetes mellitus, diagnosed for at least one year. Parental perceptions of their child's competence were assessed, together with parental perceptions of their own self-efficacy in managing their child's diabetes. Glycaemic control was assessed by the average annual HbA1C level. RESULTS: The response rate was 64.5% (51 parents); 82% were mothers and the socioeconomic class and ethnicity spread was representative of the general population. The mean age of the children was 10 years and duration of diabetes 4.4 years. Poorer glycaemic control was associated with higher perceived child competence, together with lower perceived age of responsibility, lower perceived seriousness, and less frequent blood tests. Higher parental self-efficacy and higher perceived child competence predicted a higher level of normalisation, as did lower perceived seriousness, a lower perceived parental responsibility for management, and a less protective style of parenting. CONCLUSION: Parents' perceptions of their children's diabetes are significantly related to glycaemic control; however, those who appear more competent at managing diabetes may overestimate their child's capabilities, leading to poorer glycaemic control. PMID- 16246853 TI - Parental consent in paediatric clinical research. AB - AIMS: To assess parental understanding and memorisation of the information given when seeking for consent to their child's participation to clinical research, and to identify the factors of significant influence on parents' decision making process. METHODS: Sixty eight parents who had been approached for enrolling their child in a clinical oncology or HIV study were asked to complete an interview. Their understanding was measured by a score which included items required to obtain a valid consent according to French legislation. RESULTS: Items that were best understood by parents were the aims of the study (75%), the risks (70%), the potential benefits to their child (83%), the potential benefits to other children (70%), the right to withdraw (73%), and voluntariness (84%). Items that were least understood were the procedures (44%), the possibility of alternative treatments (53%), and the duration of participation (39%). Less than 10% of the parents had understood all these points. Ten parents (15%) did not remember that they had signed up for a research protocol. Thirty three parents (48%) reported no difficulty in making their decision. Twenty four parents (38%) declared that they made their decision together with the investigator; 26 (41%) let the physician decide. Fifty four parents (78%) felt that the level of information given was satisfactory. CONCLUSION: There was an apparent discrepancy between parents' evaluation of the adequacy of the information delivered and evaluation of their understanding and memorisation. The majority of parents preferred that the physician take as much responsibility as possible in the decision making process. PMID- 16246855 TI - Harpin modulates the accumulation of salicylic acid by Arabidopsis cells via apoplastic alkalization. AB - It is reported here that salicylic acid (SA) is rapidly taken up by Arabidopsis cells, and its uptake is accompanied by media alkalization and cytosolic acidification, and it is inhibited by the ionophore nigericin, suggesting that its import is linked with that of H+ and driven by a proton gradient. Such import and accumulation declined sharply within a narrow physiological pH range (pH 5.7 6.1), corresponding to a reduction in the [H+] of the media from 1.99 micromol l( 1) to 0.79 micromol l(-1). Following the initial uptake, SA was exported back into the media as free SA against a continued [H+]-dependent import. Since the uptake and accumulation of SA declines sharply within a narrow pH range and cell wall alkalization is an early response during incompatible plant/pathogen interactions, the bacterial elicitor harpin(Pss) was used to investigate how SA transport may be modulated during defence responses. Harpin induced a rapid and sustained alkalization of the cell suspension media, reaching the critical pH (pH 5.9-6.1) at which SA import is inhibited at c. 60 min. Such media alkalization corresponded with a reduction in the SA associated with cells co-treated with harpin, and an inhibition of SA uptake in cells pretreated with harpin. Scavengers of ROS, or compounds which generate H2O2 or NO had little effect on the import or net export of SA, suggesting that media alkalization induced by harpin is sufficient to modulate the kinetics of SA transport. PMID- 16246856 TI - Cucurbit phloem serpins are graft-transmissible and appear to be resistant to turnover in the sieve element-companion cell complex. AB - Serpins are unique inhibitors of serine proteases that are located in various plant tissues and organs. An orthologue of the pumpkin (Cucurbita maxima) phloem serpin CmPS-1 was amplified from cucumber (Cucumis sativus) RNA by RT-PCR, cloned, and designated as CsPS-1 (GenBank accession no. AJ866989). Alternative amino acid sequences in the reactive centre loop suggest distinct inhibitory specificity between CmPS-1 and CsPS-1. A difference in the electrophoretic mobility of these serpins was used in heterografts to establish that serpins are phloem-mobile. Immuno light microscopy revealed that the phloem serpins are localized exclusively to sieve elements (SE), while the phloem filament protein CmPP1, used as a reference, is localized to both SEs and companion cells (CCs). Similar to CmPS-1, CsPS-1 accumulates over time in phloem exudates, indicating that serpins differ from other phloem-mobile proteins whose concentrations appear to be stable in phloem exudates. These differences could reflect alternative mechanisms regulating protein turnover and/or inaccessibility of protein degradation. The functionality of the pore/plasmodesma units connecting SEs and CCs was tested with graft-transmitted CmPP1 as a transport marker. The occurrence of CmPP1 in the CCs of the Cucumis graft partner shows that translocated 88 kDa phloem filament protein monomers can symplasmically exit the SE and accumulate in the CC. By contrast, serial sections probed with the serpin antibody demonstrate that the 43 kDa serpin does not enter CCs. Collectively, these data indicate that CCs play a decisive role in homeostasis of exudate proteins; proteins not accessing the CCs accumulate in SEs and display a time-dependent increase in concentration. PMID- 16246857 TI - Water stress-induced modifications of leaf hydraulic architecture in sunflower: co-ordination with gas exchange. AB - The hydraulic architecture, water relationships, and gas exchange of leaves of sunflower plants, grown under different levels of water stress, were measured. Plants were either irrigated with tap water (controls) or with PEG600 solutions with osmotic potential of -0.4 and -0.8 MPa (PEG04 and PEG08 plants, respectively). Mature leaves were measured for hydraulic resistance (R(leaf)) before and after making several cuts across minor veins, thus getting the hydraulic resistance of the venation system (R(venation)). R(leaf) was nearly the same in controls and PEG04 plants but it was reduced by about 30% in PEG08 plants. On the contrary, R(venation) was lowest in controls and increased in PEG04 and PEG08 plants as a likely result of reduction in the diameter of the veins' conduits. As a consequence, the contribution of R(venation) to the overall R(leaf) markedly increased from controls to PEG08 plants. Leaf conductance to water vapour (g(L)) was highest in controls and significantly lower in PEG04 and PEG08 plants. Moreover, g(L) was correlated to R(venation) and to leaf water potential (psi(leaf)) with highly significant linear relationships. It is concluded that water stress has an important effect on the hydraulic construction of leaves. This, in turn, might prove to be a crucial factor in plant-water relationships and gas exchange under water stress conditions. PMID- 16246858 TI - Root-ABA1, a major constitutive QTL, affects maize root architecture and leaf ABA concentration at different water regimes. AB - Near-isogenic hybrids (NIHs), developed from crossing maize (Zea mays L.) backcross-derived lines (BDLs) differing for the parental alleles at a major QTL for leaf ABA concentration (L-ABA), were field-tested for 2 years under well watered and water-stressed conditions. Differences among NIHs for L-ABA and other morpho-physiological traits were not affected by water regimes. On average, the QTL allele for high L-ABA markedly reduced stomatal conductance and root lodging. To elucidate the effects of the QTL on root architecture and L-ABA, root traits of two pairs of BDLs were measured in plants grown in soil columns at three water regimes. Differences among BDLs were not affected by water regimes. Across water regimes, the QTL confirmed its effect on L-ABA and showed a concurrent effect on root angle, branching, number, diameter, and dry weight. Based on these results, it is concluded that the QTL affects root lodging through a constitutive effect on root architecture. In addition, there is speculation that the QTL effects on root traits and L-ABA are probably due to pleiotropy rather than linkage and a model is proposed in which the QTL has a direct effect on root architecture, while indirectly affecting L-ABA. PMID- 16246859 TI - Bird flu and pandemic flu. PMID- 16246860 TI - Excisional surgery versus ablative surgery for ovarian endometriomata: a Cochrane Review. AB - BACKGROUND: The objective of this review was to determine which is the most effective technique for treating an ovarian endometrioma; excision or ablation. METHODS: A systematic review employing the principles of the Cochrane Menstrual Disorders and Subfertility Group was undertaken. No randomized studies of the management of endometriomata by laparotomy were found. Two randomized studies of the laparoscopic management of ovarian endometriomata of >3 cm in size were included. RESULTS: Laparoscopic excision of the cyst wall of the endometrioma was associated with a reduced rate of recurrence of the endometrioma [odds ratio (OR) 0.41, confidence interval (CI) 0.18-0.93], reduced requirement for further surgery (OR 0.21, CI 0.05-0.79), reduced recurrence rate of the symptoms of dysmenorrhoea (OR 0.15, CI 0.06-0.38), dyspareunia (OR 0.08, CI 0.01-0.51) and non-menstrual pelvic pain (OR 0.10, CI 0.02-0.56). It was also associated with a subsequently increased rate of spontaneous pregnancy in women who had documented prior subfertility (OR 5.21, CI 2.04-13.29). CONCLUSIONS: There is some evidence that excisional surgery for endometriomata provides for a more favourable outcome than drainage and ablation, with regard to the recurrence of the endometrioma, recurrence of symptoms and subsequent spontaneous pregnancy in women who were previously subfertile. Consequently this should be the favoured surgical approach. However, we found no data to indicate the best surgical approach in women planning to undergo assisted reproductive techniques. PMID- 16246861 TI - Luteal support post GnRH agonist trigger: do not stop too soon. PMID- 16246862 TI - GnRH agonist (buserelin) or HCG for ovulation induction in GnRH antagonist IVF/ICSI cycles: a prospective randomized study. PMID- 16246863 TI - Egg-sharing in return for subsidized fertility treatment: a possible solution for therapeutic cloning? PMID- 16246864 TI - Protein composition of the intranuclear inclusions of FXTAS. AB - Fragile X-associated tremor/ataxia syndrome (FXTAS) is a late-onset neurodegenerative disorder caused by premutation expansions (55-200 CGG repeats) in the fragile X mental retardation 1 (FMR1) gene. The pathologic hallmark of FXTAS is the ubiquitin-positive intranuclear inclusion found in neurons and astrocytes in broad distribution throughout the brain. The pathogenesis of FXTAS is likely to involve an RNA toxic gain-of-function mechanism, and the FMR1 mRNA has recently been identified within the inclusions. However, little is known about the proteins that mediate the abnormal cellular response to the expanded CGG repeat allele. As one approach to identify the protein mediators, we have endeavoured to define the protein complement of the inclusion itself. Fluorescence-activated flow-based methods have been developed for the efficient purification of inclusions from the post-mortem brain tissue of FXTAS patients. Mass spectrometric analysis of the entire protein complement of the isolated inclusions, combined with immunohistochemical analysis of both isolated nuclei and tissue sections, has been used to identify inclusion-associated proteins. More than 20 inclusion-associated proteins have been identified on the basis of combined immunohistochemical and mass spectrometric analysis, including a number of neurofilaments and lamin A/C. There is no dominant protein species in the inclusions, and ubiquitinated proteins represent only a minor component; thus, inclusion formation is not likely to reflect a breakdown in proteasomal degradation of nuclear proteins. The list of proteins includes at least two RNA binding proteins, heterogeneous nuclear ribonucleoprotein A2 and muscle blind like protein 1, which are possible mediators of the RNA gain-of-function in FXTAS. PMID- 16246866 TI - Brain motor system function after chronic, complete spinal cord injury. AB - Most therapies under development to restore motor function after spinal cord injury (SCI) assume intact brain motor functions. To examine this assumption, 12 patients with chronic, complete SCI and 12 controls underwent functional MRI during attempted, and during imagined, right foot movement, each at two force levels. In patients with SCI, many features of normal motor system function were preserved, however, several departures from normal were apparent: (i) volume of activation was generally much reduced, e.g. 4-8% of normal in primary sensorimotor cortex, in the setting of twice normal variance in signal change; (ii) abnormal activation patterns were present, e.g. increased pallido thalamocortical loop activity during attempted movement and abnormal processing in primary sensorimotor cortex during imagined movement; and (iii) modulation of function with change in task or in force level did not conform to patterns seen in controls, e.g. in controls, attempted movement activated more than imagined movement did within left primary sensorimotor cortex and right dorsal cerebellum, while imagined movement activated more than attempted movement did in dorsolateral prefrontal cortex and right precentral gyrus. These modulations were absent in patients with SCI. Many features of brain motor system function during foot movement persist after chronic complete SCI. However, substantial derangements of brain activation, poor modulation of function with change in task demands and emergence of pathological brain events were present in patients. Because brain function is central to voluntary movement, interventions that aim to improve motor function after chronic SCI likely also need to attend to these abnormalities of brain function. PMID- 16246865 TI - Factors affecting the clinical outcome after neural transplantation in Parkinson's disease. AB - Intrastriatal grafts of embryonic mesencephalic tissue can survive in the brains of patients with Parkinson's disease, but the degree of symptomatic relief is highly variable and some cases develop troublesome dyskinesias. Here we explored, using clinical assessment and 18F-dopa and 11C-raclopride PET, factors which may influence the functional outcome after transplantation. We observed increased 18F dopa uptake in the grafted putamen, signifying continued survival of the transplanted dopaminergic neurons, in parallel with a progressive reduction of 18F-dopa uptake in non-grafted regions for the whole patient group. The patients with the best functional outcome after transplantation exhibited no dopaminergic denervation in areas outside the grafted areas either preoperatively or at 1 or 2 years post-operatively. In contrast, patients with no or modest clinical benefit showed reduction of 18F-dopa in ventral striatum prior to or following transplantation, which may have limited graft-induced improvement. We obtained no evidence that dyskinesias were caused by abnormal dopamine (DA) release from the grafts. As has been observed for intrinsic dopaminergic neurons, there was a significant correlation between 18F-dopa uptake and methamphetamine-induced change of 11C-raclopride binding (as a measure of DA release) in the putamen containing the graft. Furthermore, we observed no correlation between 11C raclopride binding in anterior, posterior or entire putamen under basal conditions or after methamphetamine, and dyskinesia severity scores in the contralateral side of the body. Withdrawal of immunosuppression at 29 months after transplantation caused no reduction of 18F-dopa uptake or worsening of UPDRS motor score, indicating continued survival and function of the graft. However, patients showed increased dyskinesia scores, which might have been caused either by growth of the graft or worsening of a low-grade inflammation around the graft. These findings indicate that poor outcome after transplantation is associated with progressive dopaminergic denervation in areas outside the grafts, a process which may have started already before surgery. Also, that the development of dyskinesias after transplantation is not associated with excessive DA release from the grafts. Finally, our data provide evidence that long-term immunosuppression can be withdrawn without interfering with graft survival or the motor recovery induced by transplantation. PMID- 16246868 TI - Neuronal nitric oxide synthase controls enzyme activity pattern of mitochondria and lipid metabolism. AB - Mitochondria are affected by endogenous nitric oxide (NO). Besides effects of NO on mitochondrial enzymes and the stimulation of mitochondrial H2O2 production, a NO-dependent increase in mitochondrial biogenesis in several tissues has been reported. It is still obscure whether NO generated by one specific or different NO synthase (NOS) isoenzymes determine such effects. Therefore, we analyzed the amount of mitochondria, respiratory chain enzyme complexes, and citrate synthase in the brain, muscle, heart, kidney, and liver by comparing wild-type (WT) mice and mice lacking the neuronal nitric oxide synthase isoform (nNOS-KO). Our results show that the activities of NADH:cytochrome c oxidoreductase and succinate cytochrome c oxidoreductase differ between WT and nNOS-KO mice. However, similar quantities of mitochondria were found in the homogenates of tissues in WT and nNOS-KO animals. Most impressive, higher activities and protein of citrate synthase were found in the brain, muscle, heart, kidney, and liver of nNOS-KO mice. Additionally, higher contents of fatty acid synthase and lipids were determined in the livers of nNOS-KO mice but not in the heart and brain. Furthermore, liver mitochondria from nNOS-KO mice consumed pyruvate at a higher rate and released more citric acid. Our data document a previously unrecognized role of endogenous NO in the regulation of lipid metabolism. PMID- 16246867 TI - Effects of epigenetic modulation on reporter gene expression: implications for stem cell imaging. AB - Tracking stem cell localization, survival, differentiation, and proliferation after transplantation in living subjects is essential for understanding stem cell biology and physiology. In this study, we investigated the long-term stability of reporter gene expression in an embryonic rat cardiomyoblast cell line and the role of epigenetic modulation on reversing reporter gene silencing. Cells were stably transfected with plasmids carrying cytomegalovirus promoter driving firefly luciferase reporter gene (CMV-Fluc) and passaged repeatedly for 3-8 months. Within the highest expressor clone, the firefly luciferase activity decreased progressively from passage 1 (843+/-28) to passage 20 (250+/-10) to passage 40 (44+/-3) to passage 60 (3+/-1 RLU/microg; P<0.05 vs. passage 1). Firefly luciferase activity was maximally rescued by treatment with 5-azacytidine (DNA methyltransferase inhibitor) compared with trichostatin A (histone deacetylase inhibitor) and retinoic acid (transcriptional activator; P<0.05). Increasing dosages of 5-azacytidine treatment led to higher levels of firefly luciferase mRNA (RT-PCR) and protein (Western blots) and inversely lower levels of methylation in the CMV promoter (DNA nucleotide sequence). These in vitro results were extended to in vivo bioluminescence imaging (BLI) of cell transplant in living animals. Cells treated with 5-azacytidine were monitored for 2 wk compared with 1 wk for untreated cells (P<0.05). These findings should have important implications for reporter gene-based imaging of stem cell transplantation. PMID- 16246869 TI - Risk assessment. PMID- 16246870 TI - With apologies to Albert: everything is not relative. PMID- 16246872 TI - Lifetime risk of suicide in people with schizophrenia lower than commonly reported. PMID- 16246871 TI - The incidence of delirium in older people with a mood disorder is similar with lithium and valproate. PMID- 16246873 TI - Attendance at accident and emergency for deliberate self harm predicts increased risk of suicide, especially in women. PMID- 16246874 TI - Early onset of drinking increases alcohol use in adulthood. PMID- 16246875 TI - The sentence completion test for depression can distinguish between people with and without major depressive disorder. PMID- 16246876 TI - Long acting injectable naltrexone is effective and safe for treating alcohol dependence. PMID- 16246877 TI - Bipolar disorder in young people: divalproex sodium no more effective than lithium for maintenance. PMID- 16246878 TI - Adding cognitive therapy to minimal psychiatric care prevents short term, but not long term, relapse in people with bipolar disorder. PMID- 16246880 TI - Some psychosocial therapies may reduce depression, aggression, or apathy in people with dementia. PMID- 16246879 TI - Cognitive behavioural therapy reduces psychological distress in carers of people with Parkinson's disease. PMID- 16246881 TI - Collaborative care depression management for older adults: level of comorbidity does not affect outcome. PMID- 16246882 TI - Collaborative care management improves physical functioning in older people with depression. PMID- 16246883 TI - St John's wort is at least as effective as paroxetine in reducing severity of depression and is better tolerated. PMID- 16246884 TI - Postnatal home visits from healthcare professionals show promise for preventing postnatal depression. PMID- 16246885 TI - Adding hypnosis to cognitive behavioural therapy may reduce some acute stress disorder symptoms. PMID- 16246887 TI - Pharmacotherapy is an effective treatment option for generalised anxiety disorder. PMID- 16246886 TI - Coordinated care consisting of cognitive behavioural therapy plus medication improves panic disorder. PMID- 16246888 TI - Methadone plus contingency management or performance feedback reduces cocaine and opiate use in people with drug addiction. PMID- 16246889 TI - Forensic database study suggests selective serotonin reuptake inhibitors do not increase the risk of suicide in people taking antidepressants. PMID- 16246890 TI - Suicide risk peaks in first week of psychiatric hospitalisation and post discharge. PMID- 16246891 TI - Atypical antipsychotic use during the first trimester of pregnancy may not increase major malformations. PMID- 16246892 TI - Comorbid pain related somatisation is common in Latin Americans with major depressive disorder. PMID- 16246893 TI - Prescription of pharmacotherapy for depression in elderly people varies with age, race, gender, and length of care. PMID- 16246894 TI - Fifteen per cent of people treated for mental health disorders are homeless. PMID- 16246895 TI - Biology of a novel organic solute and steroid transporter, OSTalpha-OSTbeta. AB - Using a comparative approach, recent studies have identified and functionally characterized a new type of organic solute and steroid transporter (OST) from skate, mouse, rat, and human genomes. In contrast to all other organic anion transporters identified to date, transport activity requires the coexpression of two distinct gene products, a predicted 340-amino acid, seven-transmembrane (TM) domain protein (OSTalpha) and a putative 128-amino acid, single-TM domain ancillary polypeptide (OSTbeta). When OSTalpha and OSTbeta are coexpressed in Xenopus oocytes, they are able to mediate transport of estrone 3-sulfate, dehydroepiandrosterone 3- sulfate, taurocholate, digoxin, and prostaglandin E2, indicating a role in the disposition of key cellular metabolites or signaling molecules. OSTalpha and OSTbeta are expressed at relatively high levels in intestine, kidney, and liver, but they are also expressed at lower levels in many human tissues. Indirect immunofluorescence microscopy revealed that intestinal OSTalpha and OSTbeta proteins are localized to the baso-lateral membrane of mouse enterocytes. In MDCK cells, mouse Ostalpha-Ostbeta mediated the vectorial movement of taurocholate from the apical to the basolateral membrane, but not in the opposite direction, indicating basolateral efflux of bile acids. Overall, these findings indicate that OSTalpha-OSTbeta is a heteromeric transporter that is localized to the basolateral membrane of specific epithelial tissues and serves to regulate the export and disposition of bile acids and structurally related compounds from the cell. If confirmed, this model would have important implications for the body's handling of various steroid-derived molecules and may provide a new pharmacologic target for altering sterol homeostasis. PMID- 16246896 TI - Copper alters the conformation and transcriptional activity of the tumor suppressor protein p53 in human Hep G2 cells. AB - The tumor suppressor protein p53 plays a role in the molecular response to DNA damage by acting as a DNA-binding transcription factor that regulates specific target genes to arrest the cell cycle, induce repair mechanisms, and initiate apoptotic cell death. To test the effect of copper on the transcriptional activity of p53, Hep G2 cells were transiently transfected with a luciferase reporter gene downstream from multiple p53 response elements. Co-transfection with the p53 gene resulted in a 6-fold increase in luciferase activity, showing that p53 acts as a transcription factor in this system. However, in the presence of copper, luciferase activity was significantly reduced. Oligonucleotide arrays representing 145 known p53-associated genes were hybridized with biotinylated cDNAs from mRNA extracted from control and copper-treated Hep G2 cells. Among the genes that were differentially regulated were fos, RB1, glutathione peroxidase, TGF-beta, and 15-lipoxygenase, a gene known to be activated by mutant p53. Although control Hep G2 cells synthesize wild-type p53, immunocytochemistry identified not only wild type, but also mutant p53 in the presence of copper and other agents that induce oxidative damage. Thus, this report not only identifies genes that may play a role in copper-mediated apoptosis, but also suggests that copper-induced oxidative processes result in the synthesis of mutant p53 with altered transcriptional properties. PMID- 16246897 TI - Effects of gpx4 haploid insufficiency on GPx4 activity, selenium concentration, and paraquat-induced protein oxidation in murine tissues. AB - Selenium-dependent glutathione peroxidase-4 (GPx4) catalyzes the reduction of phospholipid hydroperoxides. Because a full gpx4 knockout is embryonic lethal, we examined the effect of deletion of one copy of gpx4 on the activities of three selenoperoxidases (GPx1, GPx3, and GPx4), selenium concentrations, and pro oxidant-induced protein oxidation in various tissues of mice. A total of 32 gpx4 hemizygous (GPx4+/-) and wild-type (WT) mice (8- to 10-weeks old; 16 males and 16 females) were fed a selenium-adequate diet and given an intraperitoneal injection of paraquat (PQ; 24 mg/kg body wt) or phosphate-buffered saline (PBS). All mice were euthanized 4 hrs after injection to collect tissues for analyses. In PBS treated mice, GPx4 activities in lung, liver, kidney, and testes of GPx4+/- mice were 24-39% lower (P < 0.05) than in WT mice. Among PQ-treated mice, only testis GPx4 activity in GPx4+/- mice was significantly lower (54% P < 0.05) than WT mice. Selenium concentration in testes, but not in other tissues, was reduced (34% P < 0.05) in GPx4+/- mice compared with WT mice, irrespective of treatment. Tissue GPx1 activities and plasma GPx3 and alanine aminotransferase (ALT) activities were unaffected by PQ treatment or gpx4 hemizygosity. Total protein carbonyl was elevated (73% P < 0.05) by PQ only in lung, and this effect of PQ was independent of genotypes. In conclusion, gpx4 haploid insufficiency reduced GPx4 activities and/or selenium concentrations, but had no effect on pro-oxidant induced protein oxidation in various tissues of mice. PMID- 16246898 TI - Influence of estrogen administration on the growth response to growth hormone (GH) in GH-deficient mice. AB - In women who are growth hormone (GH) deficient, exogenous estrogens increase the dosage of GH that is needed to normalize circulating levels of insulin-like growth factor (IGF-1). Serum IGF-1 derives mostly from the liver, and it is unknown whether the peripheral effects of GH are also impaired by estrogens. Because the ultimate effect of GH is longitudinal growth, we have investigated the influence of estrogen administration on the growth response to recombinant mouse GH therapy in prepubertal GH-deficient (GHD) GHRH knockout (GHRHKO) female mice. Twenty-four GHRHKO female mice (4 animals/group) were treated for 4 weeks (from the second to sixth week of age) with the following schedules: Group I, GH only (25 microg/day); Group II, subcutaneous (sc) ethynil estradiol (EE) (0.035 ES01247g/day); Group III, GH + scEE; Group IV, oral (po) EE (0.035 microg/day); Group V, GH + poEE; Group VI, placebo. At the end of the treatment period, we measured uterine weight, total body weight (TBW), body length (nose-anus, N-A), and femur length. In addition, serum IGF-1 levels were measured. Uteri of mice treated with oral or scEE showed similar increases in weight. There was no difference in the increase in longitudinal growth parameters between mice treated with GH alone or with GH in association with oral or scEE. Serum IGF-1 decreased in animals treated with GH + scEE, compared with GH group, but no group was significantly different from placebo. These results show that subcutaneous or oral EE does not reduce the growth response to GH in female GHD mice. PMID- 16246899 TI - Localization of fertility factor SP22 to specific cell types within the anterior pituitary gland. AB - Sperm protein 22 (SP22) was recently identified in the anterior pituitary gland (AP) of male Golden Syrian hamsters using ion trap mass spectrometry. SP22 has been implicated in apoptosis, androgen receptor function, fertility, and ontogeny of early-onset Parkinson's disease. However, the role of SP22 in the pituitary has not been investigated. We cloned the cDNA for full-length SP22 from AP and posterior lobe (posterior pituitary and intermediate lobe) of the pituitary gland in adult male rats and Golden Syrian hamsters, confirming the presence of SP22 mRNA in the AP and posterior lobe. Because gonadal steroids are important regulators of AP function, and SP22 is associated with androgen receptor function, we used Western blots to compare SP22 in the AP of intact and orchidectomized male rats given placebo or a low or high dose of testosterone. SP22 did not differ with treatment, indicating that AP SP22 concentration was not regulated by testosterone. To localize SP22 to specific cells of the AP, mirror image paraffin sections were labeled against SP22 and either luteinizing hormone (LH)beta, thyroid-stimulating hormone (TSH)beta, prolactin, adrenocorticotropic hormone (ACTH), or growth hormone (GH) using peroxidase-conjugated secondary antibody. Additional sections were colabeled with SP22 and one of the AP hormones using fluorescent secondary antibodies. SP22 colocalized in somatotropes and thyrotropes in rat and hamster. We identified SP22 in a small percentage of corticotropes, gonadotropes, and lactotropes. This is the first report that SP22 mRNA is present specifically in the AP, and SP22 is localized primarily in somatotropes and thyrotropes. SP22 may help regulate AP function and be particularly important for the control of GH and TSH secretion. PMID- 16246900 TI - The cross talk between protein kinase A- and RhoA-mediated signaling in cancer cells. AB - The cross talk between cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) and RhoA-mediated signal transductions and the effect of this cross talk on biologic features of human prostate and gastric cancer cells were investigated. In the human gastric cancer cell line, SGC-7901, lysophosphatidic acid (LPA) increased RhoA activity in a dose-dependent manner. The cellular permeable cAMP analog, 8-chlorophenylthio-cAMP (CPT-cAMP), inhibited the LPA-induced RhoA activation and caused phosphorylation of RhoA at serine(188). Immunofluorescence microscopy, Western blotting, and green fluorescent protein (GFP)-tagged RhoA location assay in live cells revealed that RhoA was distributed in both the cytoplasm and nucleus of SGC-7901 cells. Treatment with LPA and/or CPT-cAMP did not induce obvious translocation of RhoA in the cells. The LPA treatment caused formation of F-actin in SGC-7901 cells, and CPT-cAMP inhibited the formation. In a modified Boyden chamber assay, LPA stimulated the migration of SGC-7901 cells, and CPT-cAMP dose-dependently inhibited the stimulating effect of LPA. In soft agar assay, LPA stimulated early proliferation of SGC-7901 cells, and CPT-cAMP significantly inhibited the growth of LPA-stimulated cells. In the prostate cancer cell line, PC-3, LPA caused morphologic changes from polygonal to round, and transfection with plasmid DNA encoding constitutively active RhoA(63L) caused a similar change. Treatment with CPT-cAMP inhibited the changes in both cases. However, in PC-3 cells transfected with a plasmid encoding mutant RhoA188A, LPA induced rounding, but CPT-cAMP could not prevent the change. Results of this experiment indicated that cAMP/PKA inhibited RhoA activation, and serine188 phosphorylation on RhoA was necessary for PKA to exert its inhibitory effect on RhoA activation. The cross talk between cAMP/PKA and RhoA-mediated signal transductions had significant affect on biologic features of gastric and prostate cancer cells, such as morphologic and cytoskeletal change, migration, and anchorage-independent growth. The results may be helpful in implementing novel therapeutic strategies for invasive and metastatic prostate and gastric cancers. PMID- 16246901 TI - Revascularization determines volume retention and gene expression by fat grafts in mice. AB - Autologous fat transplantation is a popular and useful technique in plastic and reconstructive surgery. The efficiency and survival of such grafts is predictable in many cases, but there are still issues to be resolved, such as how to improve graft volume retention. To address the issue of volume retention, we studied the effect of revascularization from the recipient on the size and function of adipocytes in fat grafts. Treatment of mice with TNP-470, an angiogenesis inhibitor, reduced blood flow from the recipient into the graft after subcutaneous transplantation of epididymal fat. The weight of transplanted tissues and the size of adipocytes in the grafts were significantly lower in mice treated with TNP-470 (TNP mice) than in control mice. Expression of genes for enzymes related to lipid accumulation was decreased in the grafts of TNP mice compared with control mice. Moreover, the expression of adipocyte-derived angiogenic peptides, VEGF and leptin, was significantly lower in the grafts of TNP mice than in grafts from control animals. The expression of VEGF and leptin by cultured human adipocytes was increased in the presence of conditioned medium from cultured vascular endothelial cells. These results show that the inhibition of the revascularization of fat grafts after transplantation reduces graft volume retention and cellular function. Early and adequate revascularization may be important for both the supply of nutrients and vasoactive interactions between vascular endothelial cells and adipocytes in graft. PMID- 16246902 TI - Bifidobacterium breve and Streptococcus thermophilus secretion products enhance T helper 1 immune response and intestinal barrier in mice. AB - Lactic acid bacteria or their secretion products can modulate immune responses differently in normal and inflammatory conditions. This comparative study analyzes the effect of oral administration of living lactic acid bacteria, or their conditioned media, on the epithelial and immune functions of colitis-prone C57BL/6 IL-10-deficient mice. Mice were untreated (control) or infected with Helicobacter hepaticus with or without oral treatment with living bacteria, Bifidobacterium breve C50 and Streptococcus thermophilus 065 (LB), or their culture-conditioned media (CM). Histology, cytokine mRNA, electrical resistance, and barrier capacity of colonic samples as well as cytokine secretion by mesenteric lymph node (MLN) cells were studied. Helicobacter hepaticus mice developed only mild colitis, which was not modified in LB or CM groups. In the CM (but not the LB) group, the colonic barrier was reinforced as compared to the other groups, as evidenced by decreased horseradish peroxidase (HRP) transcytosis and mannitol fluxes and increased electrical resistance. In MLN, the percentage of CD4+ and CD8+ T cells secreting IFNgamma was significantly higher in CM (2.06% and 1.98%, respectively) mice than in H. hepaticus (1.1% and 0.47%, P < 0.05) or control mice. In addition, the nonspecific stimulation of IFNgamma, TNFalpha, and IL-12 secretion by MLN cells was significantly higher in the CM group as compared to the other groups. In the absence of severe colitis, Bifidobacterium breve C50- and Streptococcus thermophilus 065-conditioned media can reinforce intestinal barrier capacity and stimulate Th1 immune response, highlighting the involvement of lactic acid bacteria-derived components in host defense. PMID- 16246903 TI - Infectivity of hepatic strain Klebsiella pneumoniae in diabetic mice. AB - Besides urinary tract infection (UTI) and pneumonia, increased severe liver abscesses caused by Klebsiella pneumoniae (KP), especially in diabetic patients, have been observed in infections acquired in hospitals. This indicates that different KP strains with higher virulence have emerged in recent years. Our goal was to investigate the infectivity of KP isolates in mice from liver abscess or UTI patients. Mice were injected with streptozotocin to induce diabetes. Male ICR mice were infected with KpU1 (UTI strain CG3 for survival experiment only) and KpL1 (liver abscess strain CG5) by tail-vein injection of 5 x 10(4) colony forming units (CFU) bacterial suspension. The mice survival rates, cytokine level by enzyme-linked immunosorbent assay (ELISA), and bacterial presence in liver tissue by Giemsa stain were examined. The survival rates for the KpL1-infected animals were 28% and 0% in normal and diabetic groups, respectively, whereas, for the KpU1-infected mice, the rates were 100% and 75% during a 30-day observation. Nonsurviving KpL1-infected mice showed > 10(5) bacteria/ml blood and the bacteria appeared in the liver sinus area and inside liver cells. The KpL1-infected mice showed a tendency to increase the blood interleukin 1beta (IL-1beta) level in both nondiabetic and diabetic groups, whereas the tumor necrosis factor-alpha (TNF-alpha) level was significantly decreased in the KpL1-infected diabetic mice (P = 0.002). In conclusion, the KP strain from liver abscess showed a greater virulence in mice than the KP from UTI and was more virulent in diabetic than in nondiabetic mice. The infection with KP from liver abscess significantly decreased the blood TNF-alpha level in diabetes mellitus (DM) mice and the blood IL-1beta level tended to increase in both infected nondiabetic and diabetic groups. High blood bacterial count and appearance of bacteria in liver sinus and cells usually contribute to death of the animals. PMID- 16246904 TI - Production of modified C-reactive protein in U937-derived macrophages. AB - Plasma C-reactive protein (CRP) has been proposed to be a strong independent predictor for cardiovascular disease. This circulating form of CRP (native CRP or nCRP) is well described. Recently, the existence of a conformationally distinct isoform of CRP (modified CRP or mCRP) has been reported. The relevance of each CRP isoform to atherosclerotic disease is unknown. The purpose of this study was to examine the natural expression of CRP in undifferentiated, differentiated, and stimulated macrophages, cells known to contribute to atherogenesis in vivo, and to determine whether transcribed CRP was expressed as nCRP or mCRP. Macrophages were generated from U937 monocytes using phorbol 12-myristate 13-acetate. Differentiated macrophages were further stimulated with lipopolysaccharides (LPS). In undifferentiated, differentiated, and stimulated cells, CRP expression was assessed by reverse transcription-polymerase chain reaction, and CRP protein production was measured by fluorescence microscopy and flow cytometry (cellular CRP) or high-sensitivity enzyme-linked immunosorbent assay (secreted CRP). CRP transcript was minimally expressed in undifferentiated cells. Expression increased markedly in macrophages during differentiation and was not affected by LPS at 24 hrs. Cellular CRP protein increased in a time-dependent manner after LPS stimulation, and this induction was mediated via interleukin (IL)-6 and IL 1beta. A small amount of secreted CRP was detected in the media of differentiated cells, but it was not significantly increased after LPS stimulation. Using specific monoclonal antibodies, our data indicate that cellular CRP is directly translated as the mCRP rather than the nCRP isomer. These results indicate that U937-derived macrophages are a good cell model to further study the production of mCRP under conditions relevant for the atherogenic process. PMID- 16246906 TI - Acute hyperglycemia induced by ketamine/xylazine anesthesia in rats: mechanisms and implications for preclinical models. AB - The effects of anesthetic agents, commonly used in animal models, on blood glucose levels in fed and fasted rats were investigated. In fed Sprague-Dawley rats, ketamine (100 mg/kg)/xylazine (10 mg/kg) (KX) produced acute hyperglycemia (blood glucose 178.4 +/- 8.0 mg/dl) within 20 min. The baseline blood glucose levels (104.8 +/- 5.7 mg/dl) reached maximum levels (291.7 +/- 23.8 mg/dl) at 120 min. Ketamine alone did not elevate glucose levels in fed rats. Isoflurane also produced acute hyperglycemia similar to KX. Administration of pentobarbital sodium did not produce hyperglycemia in fed rats. In contrast, none of these anesthetic agents produced hyperglycemia in fasted rats. The acute hyperglycemic effect of KX in fed rats was associated with decreased plasma levels of insulin, adrenocorticotropic hormone (ACTH), and corticosterone and increased levels of glucagon and growth hormone (GH). The acute hyperglycemic response to KX was dose dependently inhibited by the specific alpha2-adrenergic receptor antagonist yohimbine (1-4 mg/kg). KX-induced changes of glucoregulatory hormone levels such as insulin, GH, ACTH, and corticosterone were significantly altered by yohimbine, whereas the glucagon levels remained unaffected. In conclusion, the present study indicates that both KX and isoflurane produce acute hyperglycemia in fed rats. The effect of KX is mediated by modulation of the glucoregulatory hormones through stimulation of alpha2-adrenergic receptors. Pentobarbital sodium did not produce hyperglycemia in either fed or fasted rats. Based on these findings, it is suggested that caution needs to be taken when selecting anesthetic agents, and fed or fasted state of animals in studies of diabetic disease or other models where glucose and/or glucoregulatory hormone levels may influence outcome and thus interpretation. However, fed animals are of value when exploring the hyperglycemic response to anesthetic agents. PMID- 16246905 TI - The choroid plexus removes beta-amyloid from brain cerebrospinal fluid. AB - beta-Amyloid (Abeta) concentration in the cerebrospinal fluid (CSF) of the brain may be regulated by the choroid plexus, which forms a barrier between blood and brain CSF. Abeta uptake from CSF was determined as its volume of distribution (V(D)) into isolated rat choroid plexus tissue. The V(D) of [125I]Abeta1-40 was corrected by subtraction of the V(D) of [14C]sucrose, a marker for extracellular space and diffusion. Abeta uptake into choroid plexus was time and temperature dependent. Uptake of [125I]Abeta was saturable. Abeta uptake was not affected by addition of transthyretin or apolipoprotein E3. In studies with primary culture monolayers of choroidal epithelial cells in Transwells, Abeta permeability across cells, corrected by [(14)C]sucrose, was greater from the CSF-facing membrane than from the blood-facing membrane. Similarly, cellular accumulation of [125I]Abeta was concentrative from both directions and was greater from the CSF-facing membrane, suggesting a bias for efflux. Overall, these results suggest the choroid plexus selectively cleanses Abeta from the CSF by an undetermined mechanism(s), potentially reducing Abeta from normal brains and the brains of Alzheimer's disease patients. PMID- 16246907 TI - The alpha(1D)-adrenergic receptor: cinderella or ugly stepsister. AB - This Perspective focuses on the alpha(1D)-adrenergic receptor (AR), the often neglected sibling of the alpha(1)-AR family. This neglect is due in part to its poor cell-surface expression. However, it has recently been shown that dimerization of the alpha(1D)-AR with either the alpha(1B)-AR or the beta(2)-AR increases alpha(1D)-AR cell-surface expression, and in this issue of Molecular Pharmacology, Hague et al. (p. 45) demonstrate that dimerization of the alpha(1D) AR with the alpha(1B)-AR not only leads to increased cell-surface expression but also results in the formation of a novel functional entity. PMID- 16246908 TI - Role of p90 ribosomal S6-kinase-1 in oltipraz-induced specific phosphorylation of CCAAT/enhancer binding protein-beta for GSTA2 gene transactivation. AB - Oltipraz, which has been extensively studied as a cancer chemopreventive agent, promotes phosphatidylinositol 3-kinase-mediated activation of CCAAT/enhancer binding protein-beta (C/EBPbeta). Activated p90 ribosomal S6-kinase-1 (RSK1) phosphorylates major transcription factors, including C/EBPbeta. This study examined whether oltipraz induces phosphorylation of C/EBPbeta at specific residues, and if so, whether RSK1 regulates C/EBPbeta phosphorylation by oltipraz for the GSTA2 gene transactivation. Subcellular fractionation and immunoblot analyses revealed that oltipraz treatment increased the level of C/EBPbeta phosphorylated at Ser(105) in the cytoplasm, which translocated to the nucleus for DNA binding in rat H4IIE cells. Immunoprecipitation-immunoblot, chromatin immunoprecipitation, and specific mutation analyses revealed that Ser(105) phosphorylated C/EBPbeta recruited the cAMP response element-binding protein binding protein for histone acetylation and transactivation of the GSTA2 gene. The role of RSK1 in Ser(105)-phosphorylation of C/EBPbeta by oltipraz and its gene transactivation was evidenced by transfection experiments with dominant negative mutants of RSK1. In mouse Hepa1c1c, human HepG2 cells, and rat primary hepatocytes, oltipraz induced phosphorylation of C/EBPbeta at Thr(217), Thr(266), and Ser(105), respectively, via RSK1. The experiment using small-interference RNA of RSK1 confirmed the essential role of RSK1 in the gene expression. Inhibition of PI3-kinase activity prevented oltipraz-inducible Ser(105)-phosphorylation of rat C/EBPbeta. Oltipraz treatment led to increases in the catalytic activity and nuclear translocation of RSK1, which was abrogated by PI3-kinase inhibition. In summary, oltipraz induces the phosphorylation of rat C/EBPbeta at Ser(105) (functionally analogous Thr(217/266) in mouse and human forms) in hepatocytes, which results in cAMP response element-binding protein-binding protein (CBP) recruitment for the GSTA2 gene transactivation, and the specific C/EBPbeta phosphorylation is mediated by RSK1 downstream of PI3-kinase. PMID- 16246909 TI - Expansion of the BioCyc collection of pathway/genome databases to 160 genomes. AB - The BioCyc database collection is a set of 160 pathway/genome databases (PGDBs) for most eukaryotic and prokaryotic species whose genomes have been completely sequenced to date. Each PGDB in the BioCyc collection describes the genome and predicted metabolic network of a single organism, inferred from the MetaCyc database, which is a reference source on metabolic pathways from multiple organisms. In addition, each bacterial PGDB includes predicted operons for the corresponding species. The BioCyc collection provides a unique resource for computational systems biology, namely global and comparative analyses of genomes and metabolic networks, and a supplement to the BioCyc resource of curated PGDBs. The Omics viewer available through the BioCyc website allows scientists to visualize combinations of gene expression, proteomics and metabolomics data on the metabolic maps of these organisms. This paper discusses the computational methodology by which the BioCyc collection has been expanded, and presents an aggregate analysis of the collection that includes the range of number of pathways present in these organisms, and the most frequently observed pathways. We seek scientists to adopt and curate individual PGDBs within the BioCyc collection. Only by harnessing the expertise of many scientists we can hope to produce biological databases, which accurately reflect the depth and breadth of knowledge that the biomedical research community is producing. PMID- 16246910 TI - Binding of herpes simplex virus-1 US11 to specific RNA sequences. AB - Herpes simplex virus-1 US11 is a RNA-binding protein with a novel RNA-binding domain. US11 has been reported to exhibit sequence- and conformation-specific RNA binding, but the sequences and conformations important for binding are not known. US11 has also been described as a double-stranded RNA (dsRNA)-binding protein. To investigate the US11-RNA interaction, we performed in vitro selection of RNA aptamers that bind US11 from a RNA library consisting of >10(14) 80 base sequences which differ in a 30 base randomized region. US11 bound specifically to selected aptamers with an affinity of 70 nM. Analysis of 23 selected sequences revealed a strong consensus sequence. The US11 RNA-binding domain and < or =46 bases of selected RNA containing the consensus sequence were each sufficient for binding. US11 binding protected the consensus motif from hydroxyl radical cleavage. RNase digestions of a selected aptamer revealed regions of both single stranded RNA and dsRNA. We observed that US11 bound two different dsRNAs in a sequence non-specific manner, but with lower affinity than it bound selected aptamers. The results define a relatively short specific sequence that binds US11 with high affinity and indicate that dsRNA alone does not confer high-affinity binding. PMID- 16246911 TI - Rearranging the centromere of the human Y chromosome with phiC31 integrase. AB - We have investigated the ability of the integrase from the Streptomyces phiC31 'phage to either delete or invert 1 Mb of DNA around the centromere of the human Y chromosome in chicken DT40 hybrid somatic cells. Reciprocal and conservative site-specific recombination was observed in 54% of cells expressing the integrase. The sites failed to recombine in the remaining cells because the sites had been damaged. The sequences of the damaged sites indicated that the damage arose as a result of repair of recombination intermediates by host cell pathways. The liability of recombination intermediates to damage is consistent with what is known about the mechanism of serine recombinase reactions. The structures of the products of the chromosome rearrangements were consistent with the published sequence of the Y chromosome indicating that the assembly of the highly repeated region between the sites is accurate to a resolution of about 50 kb. Mini chromosomes lacking a centromere were not recovered which also suggested that neo centromere formation occurs infrequently in vertebrate somatic cells. No ectopic recombination was observed between a phiC31 integrase attB site and the chicken genome. PMID- 16246912 TI - Selection of optimal oligonucleotide probes for microarrays using multiple criteria, global alignment and parameter estimation. AB - The oligonucleotide specificity for microarray hybridization can be predicted by its sequence identity to non-targets, continuous stretch to non-targets, and/or binding free energy to non-targets. Most currently available programs only use one or two of these criteria, which may choose 'false' specific oligonucleotides or miss 'true' optimal probes in a considerable proportion. We have developed a software tool, called CommOligo using new algorithms and all three criteria for selection of optimal oligonucleotide probes. A series of filters, including sequence identity, free energy, continuous stretch, GC content, self-annealing, distance to the 3'-untranslated region (3'-UTR) and melting temperature (T(m)), are used to check each possible oligonucleotide. A sequence identity is calculated based on gapped global alignments. A traversal algorithm is used to generate alignments for free energy calculation. The optimal T(m) interval is determined based on probe candidates that have passed all other filters. Final probes are picked using a combination of user-configurable piece-wise linear functions and an iterative process. The thresholds for identity, stretch and free energy filters are automatically determined from experimental data by an accessory software tool, CommOligo_PE (CommOligo Parameter Estimator). The program was used to design probes for both whole-genome and highly homologous sequence data. CommOligo and CommOligo_PE are freely available to academic users upon request. PMID- 16246915 TI - Longitudinal investigation of the relationship among maternal victimization, depressive symptoms, social support, and children's behavior and development. AB - This article is a longitudinal investigation of the relationships between maternal victimization, maternal functioning, and children's behavior and development. Participants include 203 mother-child dyads from a low-income population recruited from pediatric primary care clinics. Data are collected when children are 4 and 8 years of age. Child outcomes are evaluated using maternal, teacher, and child self-report and objective measures of cognitive and academic functioning. Maternal victimization history is associated with maternal depressive symptoms, low levels of maternal social support, child internalizing and externalizing behavior problems, and low levels of child socialization. Child behavior problems identified at age 4 are enduring and persist to age 8. The effects of maternal victimization history on both internalizing behavior problems and socialization are mediated by maternal depressive symptoms. The effects of maternal depressive symptoms on externalizing behavior problems, socialization, and anger are mediated by maternal social support. Clinical implications of findings are discussed. PMID- 16246916 TI - Differences among sexually abused and nonabused youth living with HIV. AB - Risk behaviors were compared between sexually abused and nonabused youth living with HIV (YLH). Abused YLH were significantly more likely to have attempted suicide, to have been admitted into an alcohol and/or drug treatment program, and to have engaged in crack cocaine use than were nonabused YLH and had a greater number of sexual partners. A significantly higher proportion of abused YLH had been incarcerated in contrast to nonabused youth. There were also significantly greater conduct problems among abused YLH. Finally, abused YLH had significantly higher scores on positive action and social-support coping styles than nonabused youth. Consistent with previous research, abused youth are at higher risk for a variety of negative outcomes and are also similar in many respects to sexually abused youth who are not HIV-positive. The high frequencies of two positive styles of coping among abused YLH were also observed. PMID- 16246913 TI - Transgene-induced CCWGG methylation does not alter CG methylation patterning in human kidney cells. AB - Several reports suggest that C(m)CWGG methylation tends not to co-exist with (m)CG methylation in human cells. We have asked whether or not methylation at CCWGG sites can influence CG methylation. DNA from cells expressing an M.EcoRII GFP fusion was actively methylated at CCWGG sites. CG methylation as measured by R.HpaII/R.MspI ratios was unchanged in cells expressing the transgene. Cloned representatives of C(m)CWGG methylated DNA often contained, or were adjacent to an ALU repeat, suggesting that M.EcoRII-GFP actively methylated gene-rich R-band DNA. The transgenic methyltransferase applied C(m)CWGG methylation to a representative human promoter that was heavily methylated at CG dinucleotides (the SERPINB5 promoter) and to a representative promoter that was essentially unmethylated at CG dinucleotides (the APC promoter). In each case, the CG methylation pattern remained in its original state, unchanged by the presence of neighboring C(m)CWGG sites. Q-PCR measurements showed that RNA expression from the APC gene was not significantly altered by the presence of C(m)CWGG in its promoter. Kinetic studies suggested that an adjacent C(m)CWGG methylation site influences neither the maintenance nor the de novo methylation activities of purified human Dnmt1. We conclude that C(m)CWGG methylation does not exert a significant effect on CG methylation in human kidney cells. PMID- 16246914 TI - Protein-DNA binding specificity predictions with structural models. AB - Protein-DNA interactions play a central role in transcriptional regulation and other biological processes. Investigating the mechanism of binding affinity and specificity in protein-DNA complexes is thus an important goal. Here we develop a simple physical energy function, which uses electrostatics, solvation, hydrogen bonds and atom-packing terms to model direct readout and sequence-specific DNA conformational energy to model indirect readout of DNA sequence by the bound protein. The predictive capability of the model is tested against another model based only on the knowledge of the consensus sequence and the number of contacts between amino acids and DNA bases. Both models are used to carry out predictions of protein-DNA binding affinities which are then compared with experimental measurements. The nearly additive nature of protein-DNA interaction energies in our model allows us to construct position-specific weight matrices by computing base pair probabilities independently for each position in the binding site. Our approach is less data intensive than knowledge-based models of protein-DNA interactions, and is not limited to any specific family of transcription factors. However, native structures of protein-DNA complexes or their close homologs are required as input to the model. Use of homology modeling can significantly increase the extent of our approach, making it a useful tool for studying regulatory pathways in many organisms and cell types. PMID- 16246917 TI - A psychometric examination of English and Spanish versions of the Revised Conflict Tactics Scales. AB - The psychometric properties of the Revised Conflict Tactics Scales (CTS2) are examined for English-speaking (n = 211) and Spanish-speaking (n = 194) Latino women. Internal consistency of total scale scores is satisfactory (Cronbach's alpha of .70 to .84). However, subscale alphas range from .46 to .80. Confirmatory factor analyses support five factors of negotiation, minor and severe psychological aggression, and minor and severe physical assault. In unconstrained two-group models, loadings are of similar magnitude across language of administration, with the exception of the Physical Assault scales. Unconstrained and constrained model comparisons show scale structure varied by language group for physical assault. Although results of this study show some comparability for English-speaking and Spanish-speaking Latinas, simply combining results across language groups may obscure important differences in rates of endorsement and patterns of responses reflecting cultural, educational, and economic differences. PMID- 16246918 TI - The impact of intimate partner violence on decisions to leave dating relationships: a test of the investment model. AB - This study explored the impact of violence exposure on investment-model constructs within a sample of college women involved in heterosexual dating relationships. Results generally supported the "common sense" hypothesis, suggesting that violence negatively impacts satisfaction for and commitment to one's relationship and is positively associated with intentions to leave. Exposure to psychological abuse uniquely impacted intentions to leave relationships above and beyond other model factors, suggesting that this may be a particularly important factor in determining college women's decisions. In a series of analyses examining the investment model within each of two groups (e.g., those exposed or not exposed to physical violence), results showed that the model predicted victimized women's decisions to leave as well as it predicted nonvictimized women's decisions. Taken together, results of this study suggest that victimized women base their relationship termination decisions on the same information as nonvictimized women do. PMID- 16246919 TI - Interpersonal violence cases reported to the police: a Nigerian study. AB - The study involves 204 cases of interpersonal assault reported to the police during a period of 1 year. The patterns of domestic violence and community violence involving friends, neighbors, and strangers are described. The most common type of violence reported to the police is community interpersonal violence in which victims are mostly females and perpetrators are mostly males and members of gangs. Cases of rape are reported, whereas spousal violence tends to be underreported. On the whole, female victims are more likely to be younger than their male counterparts, whereas the male perpetrators are more likely to be unemployed. Repeat assault is found in 43 instances. Possible psychological and sociological explanations are offered for violent gang activities. The need to sensitize the police to become more involved is emphasized. The author proposes suggestions that may help reduce the incidence of community violence. PMID- 16246921 TI - Criminal history and assaults on intimate partners by Mexican American and non Mexican White college students. AB - This study analyzed a sample of 348 college students to examine the role that criminal history and Mexican ethnicity play in predicting intimate partner violence. Respondents who committed crimes in the past (before the age of 15) had a higher probability of severely physically assaulting a partner than those respondents who had committed crime later in life (after the age of 15). A history of property crime was found to be a better predictor of severe partner assault than a history of violent crime. The results support a generalist perspective on crime, which states that most individuals typically do not commit one type of crime solely but commit a variety of different crimes (property and violent). This study also found differences between Mexican American and non Mexican White students in reference to minor assaults on a partner. PMID- 16246920 TI - Cognitive-behavioral rehabilitation for high-risk violent offenders: an outcome evaluation of the violence prevention unit. AB - Rehabilitation programs for adult violent offending are still novel, and few published studies examine the recidivism outcomes of those who complete such programs. This study describes a New Zealand prison program for high-risk violent men. The program is intensive and cognitive behavioral. Preliminary outcome data are presented for three indices during 2 or more years of follow-up: nonviolent reconviction, violent reconviction, and subsequent imprisonment. In comparison with untreated offenders, treated men were less likely to be reconvicted of a violent offense, and those who were took longer to fail. There was also a 12% difference in favor of the treated men on the two other indices, nonviolent reconviction and reimprisonment. The authors conclude that the program shows early promise and that further evaluation with a larger sample of treated men will be important in clarifying whether the program is having a differential impact on violent versus nonviolent offending. PMID- 16246922 TI - The role of shame, anger, and affect regulation in men's perpetration of psychological abuse in dating relationships. AB - This study determines the relationships between shame, anger, and men's perpetration of psychological abuse in dating relationships. The authors hypothesize the connection between shame proneness and men's use of psychological abuse with a dating partner, with anger's mediating in this relationship. In addition, the authors hypothesize that affect regulation would moderate the relationship between anger and men's use of psychological abuse. Results indicate that shame proneness and use of psychological abuse are significantly related and that anger mediates the relationship. However, affect regulation does not moderate the effects of anger on men's use of psychological abuse. These findings are consistent with theoretical conceptualizations of shame and have implications for intervention and treatment programs for perpetrators of psychological abuse in dating relationships. PMID- 16246923 TI - Child physical abuse and self-perceived social isolation among adolescents. AB - This research examines the connection between physical abuse and social isolation. Using data from the National Youth Survey, a measure of self-perceived social isolation was constructed indicating the extent to which respondents feel detached from their friends and from school. Those who had experienced violence were predicted to be more isolated than those who had not. Results strongly supported the hypothesis, controlling for theoretically relevant variables. Explanation is provided in terms of damage to attachment skills, social competence, and self-esteem concomitant to being a victim of abuse. Males were more socially isolated than females, and Hispanics more than Whites. Children with involved parents were less socially isolated; those whose parents experienced normlessness were more isolated. Children who recently experienced a stressful event or were from riskier neighborhoods were more isolated. The number of children in the family was positively related to isolation. Social isolation decreases between seventh and eighth grades. PMID- 16246925 TI - The host cytokine responses and protective immunity in oropharyngeal candidiasis. AB - Over the last three decades, the prevalence of oropharyngeal fungal infections has increased enormously, mainly due to an increasing population of immunocompromised patients, including individuals with HIV infection, transplant recipients, and patients receiving cancer therapy. The vast majority of these infections are caused by Candida species. The presence of cytokines in infected tissues ultimately dictates the host defense processes that are specific to each pathogenic organism. During oral infection with Candida, a large number of pro inflammatory and immunoregulatory cytokines are generated in the oral mucosa. The main sources of these cytokines are oral epithelial cells, which maintain a central role in the protection against fungal organisms. These cytokines may drive the chemotaxis and effector functions of innate and/or adaptive effector cells, such as infiltrating neutrophils and T-cells in immunocompetent hosts, and CD8(+) T-cells in HIV(+) hosts. Epithelial cells also have direct anti-Candida activity. Several studies have provided a potential link between lower levels of certain pro-inflammatory cytokines and susceptibility to oral C. albicans infection, suggesting that such cytokines may be involved in immune protection. The exact role of these cytokines in immune protection against oropharyngeal candidiasis is still incompletely understood and requires further investigation. Identification of such cytokines with the ability to enhance anti-fungal activities of immune effector cells may have therapeutic implications in the treatment of this oral infection in the severely immunocompromised host. PMID- 16246926 TI - Hereditary blood coagulation disorders: management and dental treatment. AB - Patients with hereditary hemostatic disorders, characterized by a tendency to bleeding or thrombosis, constitute a serious challenge in the dental practice. Advances in the medical diagnosis of hemostatic disorders have exposed dental professionals to new patients not amenable to the application of the management protocols associated with other, more well-known, disorders. It is the aim of this paper to review the evidence, to highlight the areas of major concern, and to suggest management regimens for patients with hereditary hemostatic disorders. An extensive review has been made (PubMed, Science Direct, Web of Knowledge, etc.) of literature pertaining to hereditary disorders affecting blood coagulation factors and how they affect the practice of dentistry. Several aspects relating to the care of such patients must be recognized and taken into consideration when dental treatment is planned. Replacement of deficient coagulation factors ensures that safe dental treatment will be carried out. However, the half-life of such coagulation factors requires that dental treatment be specifically planned and adapted to the type of pathology involved. PMID- 16246927 TI - Making the cut in dental enamel--the discovery of enamelysin (MMP-20). PMID- 16246928 TI - Bioavailability of fluoride in drinking water: a human experimental study. AB - It has been suggested that systemic fluoride absorption from drinking water may be influenced by the type of fluoride compound in the water and by water hardness. Using a human double-blind cross-over trial, we conducted this study to measure c(max), T(max), and Area Under the Curve (AUC) for plasma F concentration against time, following the ingestion of naturally fluoridated hard and soft waters, artificially fluoridated hard and soft waters, and a reference water. Mean AUC over 0 to 8 hours was 1330, 1440, 1679, 1566, and 1328 ng F.min.mL(-1) for naturally fluoridated soft, naturally fluoridated hard, artificially fluoridated soft, artificially fluoridated hard, and reference waters, respectively, with no statistically significant differences among waters for AUC, c(max), or T(max). Any differences in fluoride bioavailability between drinking waters in which fluoride is present naturally or added artificially, or the waters are hard or soft, were small compared with large within- and between subject variations in F absorption. Abbreviations used: F, fluoride; AUC, Area under the Curve for plasma F concentration against time; AUC(0-3), Area under the Curve for plasma F concentration against time for 0 to 3 hours following water ingestion; AUC(0-8), Area under the Curve for plasma F concentration against time for 0 to 8 hours following water ingestion; c(max), maximum plasma F concentration corrected for baseline plasma F and dose (i.e., F concentration of individual waters); T(max), time of c(max). PMID- 16246930 TI - VIP Inhibits Porphyromonas gingivalis LPS-induced immune responses in human monocytes. AB - Lipopolysaccharide (LPS) from the Gram-negative pathogen Porphyromonas gingivalis (Pg) stimulates cytokine secretion in immune cells, and thereby initiates the inflammation associated with periodontitis. Modulation of pro-inflammatory cytokine activity is a plausible therapeutic target in periodontal disease. Vasoactive intestinal peptide (VIP) has a role in immunoregulation, and has been identified as a molecule with therapeutically beneficial immunosuppressive effects in inflammatory and autoimmune conditions. We aimed to investigate the effect of VIP on immune responses induced by Pg LPS in vitro. VIP (10(-8) M) significantly (P < 0.05) inhibits TNF-alpha production by human monocytic THP1 cells stimulated with Pg LPS. In parallel, we showed that VIP inhibits nuclear translocation of NFkappaB and c-Jun in a time-dependent manner, but does not decrease the expression of CD14 receptors. This is the first report to show the potential of VIP as an immunomodulator of Pg-stimulated inflammatory pathways in human monocytes. PMID- 16246929 TI - NF-kappaB activation in human dental pulp stem cells by TNF and LPS. AB - Post-natal human dental pulp stem cells (DPSCs) represent a unique precursor population in the dental pulp, which has multipotential and can regenerate a dentin/pulp-like structure. Because the dental pulp is frequently infected by oral bacteria due to dental decay, in this study, we examined whether lipopolysaccharide (LPS) and tumor necrosis factor (TNF) activated the immunologic transcription factor nuclear factor kappa B (NF-kappaB) in DPSCs. We found that both TNF and LPS activated the I-kappa B kinase complex (IKK) in DPSCs to induce the phosphorylation and degradation of IkappaBalpha, resulting in the nuclear translocation of NF-kappaB. Consistently, both TNF and LPS rapidly induced the expression of the NF-kappaB-dependent gene interleukin-8 (IL-8). However, unlike in monocytes, we found that LPS could not induce the phosphorylation of the NF-kappaB active subunit p65 in DPSCs. In summary, our studies suggest that DPSCs may be involved in immune responses during pulpal infection through activating NF-kappaB. PMID- 16246931 TI - Induction of MMP-3 by hyaluronan oligosaccharides in temporomandibular joint chondrocytes. AB - Low-molecular-weight hyaluronan (LMW-HA) is often increased in osteoarthritic joints; however, its biological function in cartilage has not been clarified. We hypothesize that LMW-HA causes the catabolic activation of chondrocytes through its interaction with CD44. Cartilage explants and chondrocytes, derived from bovine temporomandibular joints (TMJ), were examined for matrix loss and the expression of matrix metalloproteinase-3 (MMP-3) following treatment with hyaluronan oligosaccharides (HA(oligos)). Hyaluronan and CD44 were uniformly distributed throughout the fibrous and cartilaginous zones of the TMJ condyle. Treatment of cartilage explants with HA(oligos) resulted in cartilage matrix loss with increased secreted caseinolytic activity. HA(oligos) treatment of TMJ chondrocytes resulted in enhanced MMP-3 expression, whereas wash-out of the HA(oligos) in the middle of the experimental period reduced this induction. These results suggest that HA(oligos) activate chondrocytes, resulting in a substantial enhancement of proteinase expression, and the removal of HA(oligos) by wash-out reverses this catabolic activation. PMID- 16246932 TI - Osteoclastogenic activity during mandibular distraction osteogenesis. AB - Mandibular distraction osteogenesis is a well-developed clinical modality for the treatment of craniofacial deformities and dental arch discrepancies, in combination with orthodontic treatment. However, in our previous study, orthodontic tooth movement into the distraction gap caused severe root resorption. The present study aimed to clarify the osteoclastogenic activity of cells in the distraction gap. We hypothesized that the gene expression of osteoclastogenic- and osteoclast-supporting molecules in osteoblasts and stromal cells would increase at distraction sites during the consolidation period. An animal model experiment involving rabbits was designed for mandibular distraction osteogenesis and subjected to in situ hybridization analysis. The number of osteoclasts was larger in the distraction gap during the early consolidation period than in normal controls, due to an increase of gene expression for osteoclastogenic cytokines in osteoblasts. It was concluded that osteoclastogenic and osteoclastic activities are stimulated at distraction sites during the early consolidation period. PMID- 16246933 TI - Apigenin and tt-farnesol with fluoride effects on S. mutans biofilms and dental caries. AB - Apigenin (Api) and tt-farnesol (Far) are two naturally occurring agents that affect the development of cariogenic biofilms. Fluoride (F) interferes physicochemically with caries development and also exhibits antibacterial activity. We examined whether the association of Api and Far enhance the anti caries properties of F by acting cooperatively on the expression of virulence of Streptococcus mutans. The biological effects of each of the agents were greatly enhanced when used in combination with F. In general, biofilms treated with Api and/or Far in combination with F displayed less biomass and fewer insoluble glucans and iodophilic polysaccharides than did those treated with the test agents alone (P < 0.05). The combination of the test agents with F was highly effective in preventing caries development in rats, especially Api+Far+F, and results were comparable with those observed with chlorhexidine + F (positive control). Results from these studies showed that apigenin and tt-farnesol may enhance the cariostatic effectiveness of fluoride. PMID- 16246934 TI - Cementum and dentin in hypophosphatasia. AB - Hypophosphatasia (HPP) often leads to premature loss of deciduous teeth, due to disturbed cementum formation. We addressed the question to what extent cementum and dentin are similarly affected. To this end, we compared teeth from children with HPP with those from matched controls and analyzed them microscopically and chemically. It was observed that both acellular and cellular cementum formation was affected. For dentin, however, no differences in mineral content were recorded. To explain the dissimilar effects on cementum and dentin in HPP, we assessed pyrophosphate (an inhibitor of mineralization) and the expression/activity of enzymes related to pyrophosphate metabolism in both the periodontal ligament and the pulp of normal teeth. Expression of nucleotide pyrophosphatase phosphodiesterase 1 (NPP1) in pulp proved to be significantly lower than in the periodontal ligament. Also, the activity of NPP1 was less in pulp, as was the concentration of pyrophosphate. Our findings suggest that mineralization of dentin is less likely to be under the influence of the inhibitory action of pyrophosphate than mineralization of cementum. PMID- 16246935 TI - X-ray microanalysis of elements in the masticatory muscle after paresis of the right masseter. AB - Muscle activity and function appear to be related to ionic concentrations in the muscle. We investigated whether muscle paresis induced by injection of Botulinum toxin A (Botox) in 16-week-old pigs over a 56-day period is associated with ionic changes in the affected muscles. Tissue samples were taken from the masseter, temporalis, medial pterygoid, and geniohyoid muscles by a standardized method and used for energy-dispersive x-ray microanalysis in an environmental scanning electron microscope. The largest increase in Na(+) was measured in the right and left sides of the masseter muscle in treated animals. Additionally, a significant elevation of Na(+) was measured in the anterior part of the temporalis muscle and in the pterygoid muscle (P < 0.05). In temporalis and pterygoid muscles, an increase in sulfur in both sides of treated pigs' heads was observed. Botox((R)) has an indirect impact on ion concentrations, resulting in changes in muscle functional capacity and adaptive compensation of paretic muscle function by other muscles. PMID- 16246936 TI - MMP20 active-site mutation in hypomaturation amelogenesis imperfecta. AB - The Amelogenesis Imperfecta (AI) are a group of clinically and genetically heterogeneous disorders that affect enamel formation. To date, mutations in 4 genes have been reported in various types of AI. Mutations in the genes encoding the 2 enamel proteases, matrix metalloproteinase 20 (MMP20) and kallikrein 4 (KLK4), have each been reported in a single family segregating autosomal recessive hypomaturation AI. To determine the frequency of mutations in these genes, we analyzed 15 Turkish probands with autosomal-recessive hypomaturation AI for MMP20 and KLK4 gene mutations. No KLK4 mutations were found. A novel MMP20 mutation (g.16250T>A) was found in one family. This missense mutation changed the conserved active-site His226 residue of the zinc catalytic domain to Gln (p.H226Q). Zymogram analysis demonstrated that this missense mutation abolished MMP20 proteolytic activity. No MMP20 mutations were found in the remaining 14 probands, underscoring the genetic heterogeneity of hypomaturation AI. PMID- 16246937 TI - Phenotype of ENAM mutations is dosage-dependent. AB - Five mutations in the ENAM gene have been found to cause hypoplastic amelogenesis imperfecta (AI), with phenotypes ranging from localized enamel pitting in carriers to severe hypoplastic AI. To determine the generality of ENAM mutations in hypoplastic AI, we sequenced the ENAM gene in ten Turkish families segregating autosomal hypoplastic AI. In two families, ENAM mutations were found. A novel nonsense mutation (g.12663C>A; p.S246X) was identified in one family segregating local hypoplastic AI as a dominant trait. Affected individuals in a second family segregating autosomal-recessive AI were compound heterozygotes for a novel insertion mutation (g.12946_12947insAGTCAGTACCAGTACTGTGTC) and a previously described insertion (g.13185_13186insAG) mutation. Heterozygous carriers of either insertion had a localized enamel-pitting phenotype. These findings substantiate that enamel phenotypes of ENAM mutations may be dose-dependent, with generalized hypoplastic AI segregating as a recessive trait and localized enamel pitting segregating as a dominant trait. PMID- 16246938 TI - CD14 and TLR4 gene polymorphisms in adult periodontitis. AB - Bacterial deposits, smoking, and host genetic factors play a major role in an individual's predisposition to periodontitis. Bacterial components are recognized by CD14 and toll-like receptor 4 (TLR4), resulting in a NF-kappaB-based inflammatory response. We hypothesized that functional CD14 and TLR4 polymorphisms contribute to periodontitis susceptibility. We aimed to investigate the occurrence of CD14-260C>T, TLR4 299Asp>Gly, and 399Thr>Ile gene polymorphisms in adult periodontititis. DNA was collected from 100 patients with severe periodontitis and from 99 periodontally healthy controls. The gene polymorphisms were determined by the PCR technique. The presence of the periodontal pathogens Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans, and whether the subjects smoked, was included in the analyses. The CD14-260T/T genotype was found in 34.0% of periodontitis patients and in 20.2% of controls. Logistic regression analysis adjusted for gender, age, smoking, and prevalence of P. gingivalis and A. actinomycetemcomitans showed an association between the CD14 260T/T genotype and periodontitis (P = 0.004, OR 3.0, 95% CI 1.4-6.9). We conclude that the CD14-260T/T genotype contributes to the susceptibility to severe periodontitis in Dutch Caucasians. PMID- 16246939 TI - Longitudinal analysis of heritability for dental caries traits. AB - The role of genetic and environmental factors on dental caries progression in young children was determined. A detailed caries assessment was performed in 2 examinations on 314 pairs of twins initially 1.5 to 8 years old. Surface-based caries prevalence rates (SBCPR) and lesion severity (LSI) were computed. Heritability estimates were calculated by SOLAR software. Analyses were performed on all ages combined and by age group (1.5-< 4; 4-6; > 6). Overall heritability estimates (H) of net increments SBCPRs were H = 30.0 (p < 0.0001), and were greatest for the youngest (H = 30.0) and oldest groups (H = 46.3). Overall LSI heritability estimates [H = 36.1 (p < 0.0001)] were also greatest for the youngest (H = 51.2) and oldest groups (H = 50.6). Similar findings were found for net increments of occlusal surfaces and deep dentinal lesions SBCPRs (H = 46.4 56.2). These findings are consistent with a significant genetic contribution to dental caries progression and severity in both emerging primary and permanent dentitions. PMID- 16246940 TI - The growth hormone receptor gene is associated with mandibular height in a Chinese population. AB - Genetic influences are important in the determination of mandibular morphology, and growth hormone receptor (GHR) is believed to have an important influence on the growth of craniofacial bone. In this study, we used quantitative trait locus methods to evaluate the relationship between craniofacial morphology and single nucleotide polymorphisms (SNPs) in GHR in an unselected healthy Chinese population. We systematically screened the 10 exons and nearby introns of GHR and identified 6 SNPs. Using 4 SNPs as markers, we studied the relationships between genotypes and craniofacial linear measurements. Individuals with the genotype CC of polymorphism I526L had a significantly greater mandibular ramus length (condylion-gonion/ articulare-gonion) than those with genotype AC or AA. Haplotype analysis showed that there were also significant differences between the long and short mandibular height groups in an extreme population. Our results indicate that the GHR gene polymorphism I526L is associated with mandibular height in the Chinese population. PMID- 16246941 TI - Chewing indicators between adults with Down syndrome and controls. AB - Down syndrome induces a neuromotor deficiency that affects the orofacial musculature, and thus could be implicated in the feeding difficulties affecting people with this disease. This study aimed to investigate the differences in chewing indicators between a group of 11 adults with Down syndrome and a group of 12 healthy subjects without Down syndrome. Chewing ability was evaluated by means of video recordings taken during a standardized meal that included 10 natural foods. The variables collected were masticatory time, number of masticatory cycles, chewing frequency, number of open masticatory cycles, and number of food refusals. There were several differences in both directions for masticatory time and number of masticatory cycles between the two groups. In addition, with the exception of puree, the group with Down syndrome had significantly lower mean chewing frequency than the reference group, and was unable to eat all the foods presented. PMID- 16246942 TI - The effect of chewing sugar-free gum on gastro-esophageal reflux. AB - Regurgitated acid entering the mouth in gastro-esophageal reflux disease can cause dental erosion. Chewing gum could induce increased swallowing frequency, thus improving the clearance rate of reflux within the esophagus. The null hypothesis of this study was that chewing gum does not have any effect on the clearance of reflux from the distal esophagus. Thirty-one subjects presenting with symptoms of reflux were given a refluxogenic meal twice and were randomly selected to chew gum for half an hour after eating the meal. Esophageal pH was measured, and pH data were analyzed and compared during the postprandial periods for 2 hrs on the 2 occasions. The median (IQ range) values for the % time pH < 4 during the postprandial period without chewing gum were 5.7 (1.7-13.5) and, with chewing gum, 3.6 (0.3-7.3), respectively (p = 0.001). Chewing sugar-free gum for half an hour after a meal can reduce acidic postprandial esophageal reflux. PMID- 16246943 TI - Double-blind study on materials testing with applied kinesiology. AB - Applied Kinesiology (AK) is a scientifically unproven method used in complementary medicine to recognize the (in)tolerance of dental materials. Test retest reliability of AK was examined. The working hypothesis was the assumption that the reliability of AK would not exceed random chance. Two dentists qualified in AK examined 112 volunteers to determine individual (in)tolerance toward two dental composite materials. After the first examination, 31 subjects were excluded from further testing. At the end of the open test phase, 34 of 81 participants had been classified as "tolerant", and seven as "intolerant" to both materials. The remaining 40 individuals showed a combination of either tolerant (to material I)/intolerant (to material II), or the reverse (n = 20 each). Retrieval rate was tested under blind conditions. In 14 cases, the results of the open and blinded tests matched, whereas in 26 cases they did not (95% confidence interval, 21%-52%; p = 0.98). This outcome confirmed our working hypothesis. PMID- 16246944 TI - Apatite/amelogenin coating on titanium promotes osteogenic gene expression. AB - Osteoblast differentiation and extracellular matrix production are pivotal processes for implant osseointegration or bone tissue engineering. We hypothesized that a biomimetic coating on titanium surfaces, consisting of apatite and amelogenin, would promote such processes. Human Embryonic Palatal Mesenchymal pre-osteoblasts were used as a model for the evaluation of cell adhesion and spreading patterns, as well as mRNA expression of certain osteoblastic gene products. Real-time PCR showed significant (p < 0.05) increase in expression of type I collagen, alkaline phosphatase, and osteocalcin from cells grown on titanium with an apatite/amelogenin composite, as compared with that from cells grown on a pure titanium or apatite coating only. Osteocalcin expression was specifically stimulated by amelogenin added to the culture media. Enhanced attachment and cell spreading were also observed. The biomimetic coating promoting cell adhesion and osteoblast differentiation may have great potential for future dental and biomedical applications. PMID- 16246945 TI - Tensile strength of mineralized/demineralized human normal and carious dentin. AB - The bond strengths of resins to caries-affected dentin are low. This could be due to weakened organic matrix. The purpose of this work was to determine if the ultimate tensile strength (UTS) of excavated carious dentin is weaker than that of normal dentin. Soft caries was excavated from extracted human molars, and the tooth was vertically sectioned into slabs. Each slab was trimmed to an hourglass shape, parallel or perpendicular to the tubule direction. Half of the specimens were mineralized, while the other half were completely demineralized in EDTA. ANOVA on ranks showed that the three-factor interactions (mineralization, caries, tubule direction) were all significant (p < 0.0001), indicating that mineralization and tubule direction gave different UTS results in normal and caries-affected dentin. No significant differences were seen between the UTS of normal and and that of caries-affected demineralized dentin in the parallel or perpendicular group. The matrix of demineralized caries-affected dentin was as strong as that of normal demineralized dentin when tested in the same direction. PMID- 16246946 TI - Functional recovery of damaged skeletal muscle through synchronized vasculogenesis, myogenesis, and neurogenesis by muscle-derived stem cells. AB - BACKGROUND: Recent studies have shown that skeletal muscle-derived stem cells (MDSCs) can give rise to several cell lineages after transplantation. However, the potential therapeutic uses of MDSCs, the functional significance of the transplanted tissue, and vasculogenesis, myogenesis, and reconstitution of other tissues have yet to be investigated in detail. In addition, the relationship between MDSCs and mesenchymal bone marrow cells is of interest. METHODS AND RESULTS: We developed a severe-damage model of mouse tibialis anterior muscle with a large deficit of nerve fibers, muscle fibers, and blood vessels. We investigated the potential therapeutic use of freshly isolated CD34+/45- (Sk-34) cells. Results showed that, after transplantation, implanted cells give rise to myogenic, vascular (pericytes, vascular smooth muscle cells, and endothelial cells), and neural (Schwann) cells, as well as contributing to the synchronized reconstitution of blood vessels, muscle fibers, and peripheral nerves, with significant recovery of both mass and contractile function after transplantation. Investigation of Sk-34 cell transplantation to the renal capsule (nonmuscle tissue) and fluorescence in situ hybridization analysis for the transplanted muscle detecting the Y chromosome revealed the intrinsic plasticity of the Sk-34 cell population. In addition, there were no donor-derived Sk-34 cells in the muscle of lethally irradiated bone marrow-transplanted animals, indicating that the Sk-34 cells were not derived from bone marrow. CONCLUSIONS: These findings indicate that freshly isolated skeletal muscle-derived Sk-34 cells are potentially useful for reconstitution therapy of the vascular, muscular, and peripheral nervous systems. These results provide new insights into somatic stem and/or progenitor cells with regard to vasculogenesis, myogenesis, and neurogenesis. PMID- 16246947 TI - Pioglitazone reduces neointima volume after coronary stent implantation: a randomized, placebo-controlled, double-blind trial in nondiabetic patients. AB - BACKGROUND: Restenosis requiring reintervention limits the long-term success after coronary stent implantation. Thiazolidinediones, like pioglitazone or rosiglitazone, are oral antidiabetic drugs with additional antirestenotic properties. In a randomized, placebo-controlled, double-blind trial, we examined the effect of 6-month pioglitazone therapy on neointima volume after coronary stenting in nondiabetic coronary artery disease patients. METHODS AND RESULTS: Fifty nondiabetic patients after coronary stent implantation were randomly assigned to pioglitazone (30 mg daily; pio) or placebo (control) treatment in addition to standard therapy, and neointima volume was assessed by intravascular ultrasound at the 6-month follow-up. Both groups were comparable with regard to baseline characteristics, angiographic lesion morphology, target vessel, and length of the stented segment. In addition, there were no statistical differences in minimal lumen diameter before and after intervention, as well as reference diameter after stent implantation. In this study population of nondiabetic patients, pio treatment did not significantly change fasting blood glucose, fasting insulin, or glycosylated hemoglobin levels, as well as lipid parameters. In contrast, pio treatment significantly reduced neointima volume within the stented segment, with 2.3+/-1.1 mm3/mm in the pio group versus 3.1+/-1.6 mm3/mm in controls (P=0.04). Total plaque volume (adventitia-lumen area) was significantly lower at follow-up in the pio group (11.2+/-3.2 mm3/mm) compared with controls (13.2+/-4.2 mm3/mm; P=0.04). Moreover, the binary restenosis rate was 3.4% in the pio group versus 32.3% in controls (P<0.01). CONCLUSIONS: Thus, 6 month treatment with pio significantly reduced neointima volume after coronary stent implantation in nondiabetic patients. These data bolster the hypothesis that antidiabetic thiazolidinediones, in addition to their metabolic effects, exhibit direct antirestenotic effects in the vasculature. PMID- 16246948 TI - Coronary stent restenosis in patients treated with cilostazol. AB - BACKGROUND: Restenosis after implantation of coronary artery stents remains a significant clinical problem. We undertook a randomized, double-blind, placebo controlled trial to determine whether cilostazol, a drug that suppresses intimal proliferation, would reduce renarrowing in patients after stent implantation in native coronary arteries. METHODS AND RESULTS: We assigned 705 patients who had successful coronary stent implantation to receive, in addition to aspirin, cilostazol 100 mg BID or placebo for 6 months; clopidogrel 75 mg daily was administered to all patients for 30 days. Restenosis was determined by quantitative coronary angiography at 6 months. The minimal luminal diameter at 6 months for cilostazol-treated patients was 1.77 mm for the analysis segment (stent plus 5-mm borders) compared with 1.62 mm in the placebo group (P=0.01). Restenosis, defined as > or =50% narrowing, occurred in 22.0% of patients in the cilostazol group and in 34.5% of the placebo group (P=0.002), a 36% relative risk reduction. Restenosis was significantly lower in cilostazol-treated diabetics (17.7% versus 37.7%, P=0.01) and in those with small vessels (23.6% versus 35.2%, P=0.02), long lesions (29.9% versus 46.6%, P=0.04), and left anterior descending coronary artery site (19.3% versus 39.8%, P=0.001). There was no difference in bleeding, rehospitalization, target-vessel revascularization, myocardial infarction, or death. CONCLUSIONS: Treatment with the drug cilostazol resulted in a significantly larger minimal luminal diameter and a significantly lower binary restenosis rate compared with placebo-treated patients. These favorable effects were apparent in patients at high risk for restenosis. PMID- 16246949 TI - Genetic predictors and remodeling of dilated cardiomyopathy in muscular dystrophy. AB - BACKGROUND: Dystrophin gene mutations cause 2 common muscular dystrophies, Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD). Both are frequently associated with dilated cardiomyopathy (DCM) and premature death. We hypothesized that early diagnosis and treatment of DCM in DMD/BMD patients would lead to ventricular remodeling and that specific dystrophin gene mutations would predict cardiac involvement. METHODS AND RESULTS: Sixty-nine boys with DMD (n=62) and BMD (n=7) (mean age, 12.9 and 13.7 years, respectively) were referred to our Cardiovascular Genetics Clinic for evaluation, including echocardiography and DNA analysis. Follow-up evaluations were scheduled yearly until the first abnormal echocardiogram indicative of DCM and quarterly thereafter. After the first abnormal echocardiogram, angiotensin-converting enzyme inhibitor or beta-blocker therapy was started. beta-Blockers were added if echocardiography showed no ventricular remodeling in angiotensin-converting enzyme inhibitor-treated patients after 3 months. DCM was diagnosed in 31 subjects (DMD, 27/62, 44%; BMD, 4/7, 57%) (mean age at onset, 15.4+/-2.8 years; range, 10.4 to 21.2 years). All 31 subjects were begun on pharmacological therapy after diagnosis. On follow-up (n=29), 2 subjects (both DMD) showed stable DCM, 8 subjects (all DMD) showed improvement, and 19 subjects (16 DMD; 3 BMD) showed normalization of left ventricular size and function (total improvement, 27/29 [93%]). DNA analysis in 47 cases (68%) revealed a significant association between DCM and exon 12 and 14 to 17 mutations, possible protection against DCM by exon 51 to 52 mutations, and a trend toward significant association between onset of DCM and exon 31 to 42 mutations. Statistical significance was based on nominal probability values. CONCLUSIONS: Early diagnosis and treatment of DCM may lead to ventricular remodeling in DMD/BMD patients. Specific dystrophin gene mutations appear to be predictive of cardiac involvement, while other mutations may protect against or inhibit development of DCM. Further studies evaluating the impact of early intervention strategies on left ventricular geometry and function in muscular dystrophy patients seem warranted. PMID- 16246950 TI - Cardiology patient page. Heart attack and stroke prevention in women. PMID- 16246951 TI - Images in cardiovascular medicine. Carotid plaque reduction after medical treatment. PMID- 16246952 TI - Images in cardiovascular medicine. Renal fibromuscular dysplasia. PMID- 16246953 TI - Images in cardiovascular medicine. Functional subclavian artery compression caused by thoracic outlet syndrome. PMID- 16246954 TI - Letter regarding article by Patti et al, "Randomized trial of high loading dose of clopidogrel for reduction of periprocedural myocardial infarction in patients undergoing coronary intervention: results from the ARMYDA-2 (Antiplatelet therapy for Reduction of MYocardial Damage during Angioplasty) study". PMID- 16246955 TI - Letter regarding article by Baicu et al, "Left ventricular systolic performance, function, and contractility in patients with diastolic heart failure". PMID- 16246956 TI - Separate but not equal: the consequences of segregated health care. PMID- 16246957 TI - Oxidative stress and the vascular wall: NADPH oxidases take center stage. PMID- 16246958 TI - Type 5 phosphodiesterase inhibition: the focus shifts to the heart. PMID- 16246959 TI - Global dynamic coupling of activation and repolarization in the human ventricle. AB - BACKGROUND: The ability to determine spatial and dynamic changes in ventricular repolarization may help to understand arrhythmogenic mechanisms in humans. We hypothesized that noncontact mapping could be used to investigate global activation-repolarization coupling in the human ventricle during steady state and premature extrastimulation. METHODS AND RESULTS: Activation-recovery intervals (ARIs) determined from reconstructed unipolar electrograms by the Ensite system were analyzed during sinus rhythm, constant pacing, spontaneous ventricular ectopic beats, and premature stimulation at intermediate and short coupling intervals in the left or right ventricle of 13 patients (6 female; mean age, 48 years) without structural myocardial disease. ARIs were measured from 32 sites in each ventricle with the use of a method validated with monophasic action potential recordings and unipolar contact electrograms. Global T-wave distribution was displayed on a 3-dimensional geometry of the ventricle, with polarities opposite to the direction of activation during steady state and premature stimulation. There was a significant inverse correlation between activation times and ARIs during sinus rhythm, ventricular ectopy, and premature stimulation (r=0.72, slope=-0.76, P<0.001). Premature stimuli at short coupling intervals flattened the regression slope compared with sinus rhythm (-0.61 versus -0.81; P=0.05), but the global pattern of repolarization was preserved. In comparison to our method, the Wyatt method of ARI measurement failed to demonstrate significant coupling between activation and repolarization (r=0.34, slope=0.19). CONCLUSIONS: Global, dynamic repolarization mapping of the human ventricle is feasible. An inverse coupling of activation and repolarization during steady state and premature stimulation may preserve electric stability in the normal ventricle. PMID- 16246960 TI - Phenotypic variability and unusual clinical severity of congenital long-QT syndrome in a founder population. AB - BACKGROUND: In the congenital long-QT syndrome (LQTS), there can be a marked phenotypic heterogeneity. Founder effects, by which many individuals share a mutation identical by descent, represent a powerful tool to further understand the underlying mechanisms and to predict the natural history of mutation associated effects. We are investigating one such founder effect, originating in South Africa in approximately ad 1700 and segregating the same KCNQ1 mutation (A341V). METHODS AND RESULTS: The study population involved 320 subjects, 166 mutation carriers (MCs) and 154 noncarriers. When not taking beta-blocker therapy, MCs had a wide range of QTc values (406 to 676 ms), and 12% of individuals had a normal QTc (< or =440 ms). A QTc >500 ms was associated with increased risk for cardiac events (OR=4.22; 95% CI, 1.12 to 15.80; P=0.033). We also found that MCs with a heart rate <73 bpm were at significantly lower risk (OR=0.23; 95% CI, 0.06 to 0.86; P=0.035). This study also unexpectedly determined that KCNQ1-A341V is associated with greater risk than that reported for large databases of LQT1 patients: A341V MCs are more symptomatic by age 40 years (79% versus 30%) and become symptomatic earlier (7+/-4 versus 13+/-9 years, both P<0.001). Accordingly, functional studies of KCNQ1-A341V in CHO cells stably expressing IKs were conducted and identified a dominant negative effect of the mutation on wild-type channels. CONCLUSIONS: KCNQ1-A341V is a mutation associated with an unusually severe phenotype, most likely caused by the dominant negative effect of the mutation. The availability of an extended kindred with a common mutation allowed us to identify heart rate, an autonomic marker, as a novel risk factor. PMID- 16246961 TI - Hybrid approaches to thoracic aortic aneurysms: the role of endovascular elephant trunk completion. AB - BACKGROUND: Thoracic aortic aneurysm affecting the arch and proximal descending thoracic aorta requires 2-stage repairs that include proximal elephant trunk graft placement and completion of thoracic or thoracoabdominal repair. The application of endovascular grafting to complete the proximal procedure avoids a thoracotomy and may improve the morbidity and mortality of the patient population at risk. METHODS AND RESULTS: A retrospective review of 399 thoracic endovascular grafts at our institution between 2000 and 2004 identified 22 patients who required elephant trunk and endovascular completion. Three patients underwent mesenteric bypass in addition to their proximal repairs. Mean follow-up was 10 months (range 1 to 42 months); there were no ruptures, and all patients returned for follow-up. Technical success was achieved in all patients. The 1-, 12-, and 24-month mortality rates (by Kaplan-Meier analysis) were 4.5%, 15.8%, and 15.8%, respectively. Caudal migration of the endograft occurred in 1 patient, and all but 2 aneurysms decreased or remained stable in size. The 2 patients with growth included a type III endoleak (which resolved after treatment) and pressurization through an expanded PTFE stentgraft. Three cases of transient paraparesis occurred (all in patients requiring mesenteric bypass or abdominal aortic aneurysm repair), and there were no paraplegias or strokes. CONCLUSIONS: Endovascular completion of elephant trunks is feasible and can be accomplished with minimal mortality. Meticulous imaging follow-up is required to detect persistent aneurysm pressurization and to verify the integrity of the repair. Improvements in implant design and delivery systems will further simplify the second-stage portion of these complex aneurysm repairs. PMID- 16246962 TI - Relation between serum phosphate level and cardiovascular event rate in people with coronary disease. AB - BACKGROUND: Higher levels of serum phosphate are associated with adverse cardiovascular outcomes, especially in the setting of overt hyperphosphatemia. Given the biological importance of phosphorus, it is plausible that higher levels of serum phosphate within the normal range may also be associated with adverse outcomes. METHODS AND RESULTS: We performed a post hoc analysis of data from the Cholesterol And Recurrent Events (CARE) study. Baseline serum phosphate levels were measured in 4127 fasting participants who were randomized to receive pravastatin 40 mg daily or placebo and followed up for a median of 59.7 months. We used Cox proportional-hazards models to examine the association between serum phosphate and adverse clinical outcomes after adjustment for potential confounders. During nearly 60 months of follow-up, 375 participants died. A significant association was noted between baseline serum phosphate level and the age-, race-, and sex-adjusted risk of all-cause death (hazard ratio per 1 mg/dL, 1.27; 95% confidence interval, 1.02 to 1.58). After categorization based on baseline phosphate level (<2.5, 2.5 to 3.4, 3.5 to 3.9, and > or =4 mg/dL) and further adjustment, a graded independent relation between phosphate and death was observed (P for trend=0.03). For instance, participants with serum phosphate > or =3.5 mg/dL had an adjusted hazard ratio for death of 1.27 (95% confidence interval, 1.02 to 1.59) compared with those with serum phosphate of <3.5 mg/dL. Higher levels of serum phosphate were also associated with increased risk of new heart failure, myocardial infarction, and the composite of coronary death or nonfatal myocardial infarction, but not the risk of stroke. CONCLUSIONS: We found a graded independent relation between higher levels of serum phosphate and the risk of death and cardiovascular events in people with prior myocardial infarction, most of whom had serum phosphate levels within the normal range. Given the ready availability and low cost of serum phosphate assays, this finding may prove clinically useful. PMID- 16246963 TI - Mortality after acute myocardial infarction in hospitals that disproportionately treat black patients. AB - BACKGROUND: African Americans are more likely to be seen by physicians with less clinical training or to be treated at hospitals with longer average times to acute reperfusion therapies. Less is known about differences in health outcomes. This report compares risk-adjusted mortality after acute myocardial infarction (AMI) between US hospitals with high and low fractions of elderly black AMI patients. METHODS AND RESULTS: A prospective cohort study was performed for fee for-service Medicare patients hospitalized for AMI during 1997 to 2001 (n=1,136,736). Hospitals (n=4289) were classified into approximate deciles depending on the extent to which the hospital served the black population. Decile 1 (12.5% of AMI patients) included hospitals without any black AMI admissions during 1997 to 2001. Decile 10 (10% of AMI patients) included hospitals with the highest fraction of black AMI patients (33.6%). The main outcome measures were 90 day and 30-day mortality after AMI. Patients admitted to hospitals disproportionately serving blacks experienced no greater level of morbidities or severity of the infarction, yet hospitals in decile 10 experienced a risk adjusted 90-day mortality rate of 23.7% (95% CI 23.2% to 24.2%) compared with 20.1% (95% CI 19.7% to 20.4%) in decile 1 hospitals. Differences in outcomes between hospitals were not explained by income, hospital ownership status, hospital volume, census region, urban status, or hospital surgical treatment intensity. CONCLUSIONS: Risk-adjusted mortality after AMI is significantly higher in US hospitals that disproportionately serve blacks. A reduction in overall mortality at these hospitals could dramatically reduce black-white disparities in healthcare outcomes. PMID- 16246964 TI - Sildenafil inhibits beta-adrenergic-stimulated cardiac contractility in humans. AB - BACKGROUND: Sildenafil inhibits phosphodiesterase 5 (PDE5A) to elevate intracellular cGMP and to induce vasodilation. This effect has led to its use for treating erectile dysfunction. Although its influence on rest heart function has appeared minimal, recent animal studies suggest that sildenafil can have potent effects on hearts stimulated by beta-adrenergic or pressure overloads. We therefore tested whether sildenafil blunts dobutamine-stimulated cardiac function in humans. METHODS AND RESULTS: Thirty-five healthy volunteers underwent a randomized, double-blind, placebo-controlled study in which cardiac function was assessed in response to dobutamine before and after oral sildenafil (100 mg, n=19) or placebo (n=16). Echo Doppler and noninvasive blood pressure data yielded load-independent contractility indexes (maximal power index and end-systolic elastance), ejection fraction, and measures of diastolic function. In the initial dobutamine test, systolic and diastolic function improved similarly in both treatment groups (eg, peak power index rose 80+/-28% in the placebo group and 82+/-31% in the sildenafil group; P=NS). However, in subjects who then received sildenafil, their second dobutamine response was significantly blunted, with peak power, ejection fraction, and end-systolic elastance changes reduced by 32+/-34%, 66+/-64%, and 56+/-63%, respectively (each P<0.001 versus the initial response). This contrasted to the placebo group, which displayed similar functional responses with both dobutamine tests. Sildenafil treatment did not significantly alter diastolic changes induced by dobutamine compared with results with placebo. CONCLUSIONS: PDE5A inhibition by sildenafil blunts systolic responses to beta adrenergic stimulation. This finding supports activity of PDE5A in the human heart and its role in modifying stimulated cardiac function. PMID- 16246965 TI - Selective activation of inflammatory pathways by intermittent hypoxia in obstructive sleep apnea syndrome. AB - BACKGROUND: Obstructive sleep apnea syndrome (OSAS), characterized by intermittent hypoxia/reoxygenation (IHR), is an independent risk factor for cardiovascular disease. We investigated the underlying molecular mechanisms of this association in a translational study. METHODS AND RESULTS: In a novel in vitro model of IHR, we used HeLa cells transfected with reporter constructs and DNA binding assays for the master transcriptional regulators of the inflammatory and adaptive pathways (NFkappaB and HIF-1, respectively) to investigate underlying transcriptional events initiated by repeated cell exposure to IHR. Furthermore, we prospectively studied 19 male OSAS patients (median apnea hypopnea frequency, 48.5 episodes per hour; interquartile range [IQR], 28.5 to 72.9) and 17 matched normal control subjects. Circulating levels of the proinflammatory cytokine tumor necrosis factor-alpha and the adaptive factor erythropoietin were assayed in all subjects at baseline and again after 6 weeks of continuous positive airway pressure therapy in patients. Full blood count was measured as part of a detailed baseline evaluation. HeLa cells exposed to IHR demonstrated selective activation of the proinflammatory transcription factor NFkappaB (P<0.001 by ANOVA), whereas the adaptive regulator HIF-1 was not activated, as demonstrated by luciferase reporter assays and DNA binding studies. Circulating tumor necrosis factor-alpha levels were higher in OSAS patients (2.56 pg/mL; IQR, 2.01 to 3.42 pg/mL) than in control subjects (1.25 pg/mL; IQR, 0.94 to 1.87; P<0.001) but normalized with continuous positive airway pressure therapy (1.24 pg/mL; IQR, 0.78 to 2.35 pg/mL; P=0.002). In contrast, erythropoietin levels were similar throughout. Furthermore, circulating neutrophil levels were higher in OSAS patients than in control subjects, whereas the hematocrit was unaltered. CONCLUSIONS: These data demonstrate selective activation of inflammatory over adaptive pathways in IHR and OSAS, which may be an important molecular mechanism of cardiovascular disease. PMID- 16246966 TI - Nox1 is involved in angiotensin II-mediated hypertension: a study in Nox1 deficient mice. AB - BACKGROUND: Increased production of reactive oxygen species (ROSs) by angiotensin II (Ang II) is involved in the initiation and progression of cardiovascular diseases. NADPH oxidase is a major source of superoxide generated in vascular tissues. Although Nox1 has been identified in vascular smooth muscle cells as a new homolog of gp91phox (Nox2), a catalytic subunit of NADPH oxidase, the pathophysiological function of Nox1-derived ROSs has not been fully elucidated. To clarify the role of Nox1 in Ang II-mediated hypertension, we generated Nox1 deficient (-/Y) mice. METHODS AND RESULTS: No difference in the baseline blood pressure was observed between Nox1(+/Y) and Nox1(-/Y). Infusion of Ang II induced a significant increase in mean blood pressure, accompanied by augmented expression of Nox1 mRNA and superoxide production in the aorta of Nox1(+/Y), whereas the elevation in blood pressure and production of superoxide were significantly blunted in Nox1(-/Y). Conversely, the infusion of pressor as well as subpressor doses of Ang II did elicit marked hypertrophy in the thoracic aorta of Nox1(-/Y) similar to Nox1(+/Y). Administration of a nitric oxide synthase inhibitor (L-NAME) to Nox1(+/Y) did not affect the Ang II-mediated increase in blood pressure, but it abolished the suppressed pressor response to Ang II in Nox1(-/Y). Finally, endothelium-dependent relaxation and the level of cGMP in the isolated aorta were preserved in Nox1(-/Y) infused with Ang II. CONCLUSIONS: A pivotal role for ROSs derived from Nox1/NADPH oxidase was suggested in the pressor response to Ang II by reducing the bioavailability of nitric oxide. PMID- 16246967 TI - Reduced mitochondrial oxidative capacity and increased mitochondrial uncoupling impair myocardial energetics in obesity. AB - BACKGROUND: Obesity is a risk factor for cardiovascular disease and is strongly associated with insulin resistance and type 2 diabetes. Recent studies in obese humans and animals demonstrated increased myocardial oxygen consumption (MVO2) and reduced cardiac efficiency (CE); however, the underlying mechanisms remain unclear. The present study was performed to determine whether mitochondrial dysfunction and uncoupling are responsible for reduced cardiac performance and efficiency in ob/ob mice. METHODS AND RESULTS: Cardiac function, MVO2, mitochondrial respiration, and ATP synthesis were measured in 9-week-old ob/ob and control mouse hearts. Contractile function and MVO2 in glucose-perfused ob/ob hearts were similar to controls under basal conditions but were reduced under high workload. Perfusion of ob/ob hearts with glucose and palmitate increased MVO2 and reduced CE by 23% under basal conditions, and CE remained impaired at high workload. In glucose-perfused ob/ob hearts, mitochondrial state 3 respirations were reduced but ATP/O ratios were unchanged. In contrast, state 3 respiration rates were similar in ob/ob and control mitochondria from hearts perfused with palmitate and glucose, but ATP synthesis rates and ATP/O ratios were significantly reduced in ob/ob, which suggests increased mitochondrial uncoupling. Pyruvate dehydrogenase activity and protein levels of complexes I, III, and V were reduced in obese mice. CONCLUSIONS: These data indicate that reduced mitochondrial oxidative capacity may contribute to cardiac dysfunction in ob/ob mice. Moreover, fatty acid but not glucose-induced mitochondrial uncoupling reduces CE in obese mice by limiting ATP production and increasing MVO2. PMID- 16246968 TI - Ethnicity and peripheral arterial disease: the San Diego Population Study. AB - BACKGROUND: Previous studies have indicated higher rates of peripheral arterial disease (PAD) in blacks than in non-Hispanic whites (NHWs), with limited information available for Hispanics and Asians. The reason for the PAD excess in blacks is unclear. METHODS AND RESULTS: Ethnic-specific PAD prevalence rates were determined in a randomly selected defined population that included 4 ethnic groups; NHWs, blacks, Hispanics, and Asians. A total of 2343 participants aged 29 to 91 years were evaluated. There were 104 cases of PAD (4.4%). In weighted logistic models with NHWs as the reference group and containing demographic factors only, blacks had a higher PAD prevalence than NHWs (OR=2.30, P<0.024), whereas PAD rates in Hispanics and Asians, although somewhat lower, were not significantly different from NHWs. Blacks had significantly more diabetes and hypertension than NHWs and a significantly higher body mass index. Inclusion of these variables and other PAD risk factors in the model did not change the effect size for black ethnicity (OR=2.34, P=0.048). A model containing interaction terms for black ethnicity and each of the other risk factors revealed no significant interaction terms, which indicates no evidence that blacks were more "susceptible" than NHWs to cardiovascular disease risk factors. CONCLUSIONS: Black ethnicity was a strong and independent risk factor for PAD, which was not explained by higher levels of diabetes, hypertension, and body mass index. There was no evidence of a greater susceptibility of blacks to cardiovascular disease risk factors as a reason for their higher PAD prevalence. Thus, the excess risk of PAD in blacks remains unexplained and requires further study. PMID- 16246969 TI - Molecular regulation of platelet-dependent thrombosis. AB - Hemostasis is a normal process preventing the sequelae of uncontrolled hemorrhage. In certain settings, these same processes cause adverse clinical events due to thrombotic occlusion of a vessel. The majority of unstable coronary syndromes result from disruption of an atherosclerotic plaque, leading to the exposure of subintimal contents, which triggers coagulation and the formation of a platelet-rich thrombus. The central role of platelet activation in the events that lead to vessel occlusion is well known. However, this process is complex and influenced by a myriad of cellular and plasma-derived mediators that regulate the balance between occlusive and nonocclusive thrombosis. PMID- 16246970 TI - Effects of salt sensitivity on neural cardiovascular regulation in essential hypertension. AB - Salt-sensitive hypertensive subjects, as defined by conventional categorical classification, exhibit alterations of autonomic cardiovascular control. The aim of our study was to explore whether, in hypertensive subjects, the degree of autonomic dysfunction and the level of salt sensitivity are correlated even when the latter is only mildly elevated and displays under-threshold values. Salt sensitivity of 34 essential hypertensive subjects was assessed on a continuous basis by the salt sensitivity index after low- and high-sodium diet. Beat-by-beat finger blood pressure was recorded after each diet period. Autonomic cardiovascular control was evaluated by spectral analysis of blood pressure and pulse interval and by assessment of spontaneous baroreflex sensitivity (sequence technique). Salt sensitivity and baroreflex sensitivity showed a negative relationship during low and high sodium intake, starting from low values of the salt sensitivity index. All spectral indexes of pulse interval, except the ratio between low- and high-frequency powers, were inversely related to salt sensitivity index after high sodium intake. In subjects with lower salt sensitivity, baroreflex sensitivity and pulse interval power in the high frequency band were higher after high sodium intake than after low sodium intake. In contrast, subjects with a higher salt sensitivity index showed lower values of baroreflex sensitivity and pulse interval power in the high-frequency band, uninfluenced by salt intake. Our results provide the first demonstration of an impairment of parasympathetic cardiac control in parallel with the increase in the degree of salt sensitivity, also in subjects who were not ranked as salt sensitive by the conventional categorical classification. PMID- 16246971 TI - Thrombophilia is significantly associated with severe preeclampsia: results of a large-scale, case-controlled study. AB - The role of thrombophilia in the pathogenesis of preeclampsia is controversial. The aim of this case-controlled study was to determine whether thrombophilia increases the risk of preeclampsia or interferes with its clinical course. A total of 808 white patients who developed preeclampsia (cases) and 808 women with previous uneventful pregnancies (controls) matched for age and parity were evaluated for inherited and acquired thrombophilia (factor V Leiden; factor II G20210A; methylenetetrahydrofolate reductase C677T; protein S, protein C, and antithrombin III deficiency; anticardiolipin antibodies; lupus anticoagulant; and hyperhomocysteinemia). Odds ratios (ORs) with 95% confidence intervals (CIs) for risk of being carriers of thrombophilia in cases compared with controls and for risk of maternal life-threatening complications and adverse perinatal outcomes in preeclamptic patients with or without thrombophilia were calculated. Women with severe preeclampsia (406 cases) had a higher risk (OR, 4.9; 95% CI, 3.5 to 6.9) of being carriers of either an inherited or acquired thrombophilic factor, except for protein S, protein C, and antithrombin deficiency. In women with mild preeclampsia (402 cases), only prothrombin and homozygous methylenetetrahydrofolate reductase gene mutations were significantly more prevalent than in the controls. Thrombophilic patients with severe preeclampsia are at increased risk of acute renal failure (OR, 1.8; 95% CI, 1.5 to 2.2), disseminated intravascular coagulation (OR, 2.7; 95% CI, 1.1 to 6.4), abruptio placentae (OR, 2.6; 95% CI, 1.2 to 6.0) and perinatal mortality (OR, 1.7; 95% CI, 1.5 to 2.2) compared with nonthrombophilic preeclamptic patients. Our study demonstrates a significant association between maternal thrombophilia and severe preeclampsia in white women. Thrombophilia also augments the risk of life threatening maternal complications and adverse perinatal outcomes in preeclamptic patients. PMID- 16246973 TI - Uric acid is as important as proteinuria in identifying fetal risk in women with gestational hypertension. AB - Gestational hypertension is differentiated into higher and lower risk by the presence or absence of proteinuria. We asked if hyperuricemia, a common finding in pregnancy hypertension, might also be an indicator of increased risk. We examined fetal outcome data from 972 pregnancies collected from 1997 to 2002 in a nested case-control study. Participants were nulliparous with no known medical complications. The frequency of preterm birth, the duration of pregnancy, frequency of small-for-gestational-age infants, and birth weight centile were determined for pregnancies assigned to 8 categories by the presence or absence of combinations of hypertension, hyperuricemia, and proteinuria. In women with gestational hypertension, hyperuricemia was associated with shorter gestations and smaller birth weight centiles and increased risk of preterm birth and small for-gestational-age infants. Hyperuricemia increased the risk of these outcomes in the presence or absence of proteinuria. Risk was also increased in a small group of women with hyperuricemia and proteinuria without hypertension. Women with only hypertension and hyperuricemia have similar or greater risk as women with only hypertension and proteinuria. Those with hypertension, proteinuria, and hyperuricemia have greater risk than those with hypertension and proteinuria alone. The risk of these outcomes increased with increasing uric acid. Hyperuricemia is at least as effective as proteinuria at identifying gestational hypertensive pregnancies at increased risk. Uric acid should be reexamined for clinical and research utility. PMID- 16246972 TI - Hydrolysis of angiotensin peptides by human angiotensin I-converting enzyme and the resensitization of B2 kinin receptors. AB - We measured the cleavage of angiotensin I (Ang I) metabolites by angiotensin I converting enzyme (ACE) in cultured cells and examined how they augment actions of bradykinin B2 receptor agonists. Monolayers of Chinese hamster ovary cells transfected to stably express human ACE and bradykinin B2 receptors coupled to green fluorescent protein (B2GFP) or to express only coupled B2GFP receptors. We used 2 ACE-resistant bradykinin analogues to activate the B2 receptors. We used high-performance liquid chromatography to analyze the peptides cleaved by ACE on cell monolayers and found that Ang 1-9 was hydrolyzed 18x slower than Ang I and &30% slower than Ang 1-7. Ang 1-7 was cleaved to Ang 1-5. Although micromol/L concentrations of slowly cleaved substrates Ang 1-7 and Ang 1-9 inhibit ACE, they resensitize the desensitized B2GFP receptors in nmol/L concentration, independent of ACE inhibition. This is reflected by release of arachidonic acid through a mechanism involving cross-talk between ACE and B2 receptors. When ACE was not expressed, the Ang 1-9, Ang 1-7 peptides were inactive. Inhibitors of protein kinase C-alpha, phosphatases and Tyr-kinase blocked this resensitization activity, but not basal B2 activation by bradykinin. Ang 1-9 and Ang 1-7 enhance bradykinin activity, probably by acting as endogenous allosteric modifiers of the ACE and B2 receptor complex. Consequently, when ACE inhibitors block conversion of Ang I, other enzymes can still release Ang I metabolites to enhance the efficacy of ACE inhibitors. PMID- 16246974 TI - Hypertension during pregnancy: a disorder begging for pathophysiological support. PMID- 16246975 TI - Multicenter phase I/II study of cetuximab with paclitaxel and carboplatin in untreated patients with stage IV non-small-cell lung cancer. AB - PURPOSE: Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors have demonstrated antitumor activity in patients with non-small-cell lung cancer (NSCLC). This study examined the safety profile of the monoclonal antibody EGFR inhibitor, cetuximab, when added to paclitaxel and carboplatin in untreated patients with stage IV NSCLC. Secondary objectives included efficacy and paclitaxel and carboplatin pharmacokinetics during cetuximab treatment. PATIENTS AND METHODS: Patients with tumor evidence of EGFR by immunohistochemistry, performance status of 0 to 2, and measurable disease received paclitaxel 225 mg/m2 with carboplatin area under the curve = 6 on day 1 every 3 weeks. Cetuximab was administered at 400 mg/m2, 1 week before paclitaxel and carboplatin, then weekly at 250 mg/m2. The regimen continued until disease progression or intolerable toxicity. RESULTS: Thirty-one of 32 enrolled patients were treated. The most common cetuximab toxicity was rash in 84% of patients (grade 3 in 13%). Pharmacokinetic sampling did not reveal an interaction between carboplatin, paclitaxel, and cetuximab. An objective response was observed in eight patients (26%). With a median follow-up of 19 months, the median time to progression was 5 months, median survival was 11 months, and the 1- and 2-year survival rates were 40% and 16%, respectively. CONCLUSION: The combination of cetuximab, paclitaxel, and carboplatin was safe and well tolerated in this population of stage IV patients. The response rate, time to progression, and median survival were slightly superior to historical controls treated with paclitaxel and carboplatin alone. A randomized phase II trial has completed accrual. PMID- 16246976 TI - Prognostic significance of tumor regression after preoperative chemoradiotherapy for rectal cancer. AB - PURPOSE: We assessed the impact of tumor regression grading (TRG) and its value in correlation to established prognostic factors in a cohort of rectal carcinoma patients treated by preoperative chemoradiotherapy (CRT). PATIENTS AND METHODS: TRG was evaluated on surgical specimens of 385 patients treated within the preoperative CRT arm of the CAO/ARO/AIO-94 trial: 50.4 Gy was delivered, fluorouracil was given in the first and fifth week, and surgery was performed 6 weeks thereafter. TRG was determined by the amount of viable tumor versus fibrosis, ranging from TRG 4 when no viable tumor cells were detected, to TRG 0 when fibrosis was completely absent. TRG 3 was defined as regression more than 50% with fibrosis outgrowing the tumor mass, TRG 2 was defined as regression less than 50%, and TRG 1 was defined basically as a morphologically unaltered tumor mass. We performed an initially unplanned, hypothesis-generating analysis with respect to the prognostic value of this TRG system. RESULTS: TRG 4, 3, 2, 1, 0 was found in 10.4%, 52.2%, 13.8%, 15.3%, and 8.3% of the resected specimens, respectively. Five-year disease-free survival (DFS) after CRT and curative resection was 86% for TRG 4, 75% for grouped TRG 2 + 3, and 63% for grouped TRG 0 + 1 (P = .006). On multivariate analysis, the pathologic T category and the nodal status after CRT were the most important independent prognostic factors for DFS. CONCLUSION: In this exploratory analysis, complete (TRG 4) and intermediate pathologic response (TRG 2 + 3) suggested improved DFS after preoperative CRT. TRG assessment should be implemented in pathologic evaluation and prospectively validated in further studies. PMID- 16246977 TI - Neoadjuvant chemotherapy with high-dose Ifosfamide, high-dose methotrexate, cisplatin, and doxorubicin for patients with localized osteosarcoma of the extremity: a joint study by the Italian and Scandinavian Sarcoma Groups. AB - PURPOSE: To explore the effect of high-dose ifosfamide in first-line treatment for patients < or = 40 years of age with nonmetastatic osteosarcoma of the extremity. PATIENTS AND METHODS: From March 1997 to September 2000, 182 patients were evaluated. Primary treatment consisted of two blocks of high-dose ifosfamide (15 g/m2), methotrexate (12 g/m2), cisplatin (120 mg/m2), and doxorubicin (75 mg/m2). Postoperatively, patients received two cycles of doxorubicin (90 mg/m2), and three cycles each of high-dose ifosfamide, methotrexate, and cisplatin (120 to 150 mg/m2). Granulocyte colony-stimulating factor support was mandatory after the high-dose ifosfamide/cisplatin/doxorubicin combination. RESULTS: No disease progression was recorded during primary chemotherapy, 164 patients (92%) underwent limb-salvage surgery, four patients (2%) underwent rotation plasty, and 11 patients (6%) had limbs amputated. Three (1.6%) patients died as a result of treatment-related toxicity, and one died as a result of pulmonary embolism after pathologic fracture. Grade 4 neutropenia and thrombocytopenia followed 52% and 31% of all courses, respectively, and mild to severe nephrotoxicity was recorded in 19 patients (10%). The median received dose-intensity compared with protocol was 0.82. With a median follow-up of 55 months, the 5-year probability of event free survival was 64% (95% CI, 57% to 71%) and overall survival was 77% (95% CI, 67% to 81%), whereas seven patients (4%) experienced local recurrence. CONCLUSION: The addition of high-dose ifosfamide to methotrexate, cisplatin, and doxorubicin in the preoperative phase is feasible, but with major renal and hematologic toxicities, and survival rates similar to those obtained with four drug regimens using standard-dose ifosfamide. Italian Sarcoma Group/Scandinavian Sarcoma Group study I showed that in a multicenter setting, more than 90% of patients with osteosarcoma of the extremity can undergo conservative surgery. PMID- 16246978 TI - Anemia in the elderly: time for new blood in old vessels? PMID- 16246979 TI - In search of evidence: is there the will and a way? PMID- 16246980 TI - Thrombolytic therapy for patients with pulmonary embolism who are hemodynamically stable but have right ventricular dysfunction: pro. PMID- 16246981 TI - Thrombolysis for pulmonary embolism in patients with right ventricular dysfunction: con. PMID- 16246984 TI - Effects of adding ribavirin to interferon to treat chronic hepatitis C infection: a systematic review and meta-analysis of randomized trials. AB - Evidence shows that a combination therapy of ribavirin plus interferon clears hepatitis C virus from the blood in about 40% of patients with chronic hepatitis C infection, but the effects on clinical outcomes are unclear. We evaluated the beneficial and harmful effects of ribavirin plus interferon vs interferon alone for treatment of patients with chronic hepatitis C infection. Randomized trials were included irrespective of blinding, language, or publication status. Trials were identified through the Cochrane Hepato-Biliary Group Controlled Trials Register, the Cochrane Library, MEDLINE, EMBASE, manual searches of bibliographies and journals, and correspondence with experts (in May 2004). Data were extracted independently by 2 reviewers. The primary outcomes were morbidity plus mortality and viral clearance. Secondary outcomes included histologic response, quality of life, and adverse events. Previous antiviral therapy (treatment-naive patients, relapsers, or nonresponders), patient characteristics, treatment regimen, methodological quality, and duration of follow-up were extracted. We included 72 trials with a total of 9991 enrolled patients. Treatment with ribavirin plus interferon significantly reduced morbidity plus mortality (Peto odds ratio, 0.46; 95% confidence interval [CI], 0.22-0.96) and significantly improved sustained viral clearance in treatment-naive patients (risk ratio, 0.72; 95% CI, 0.68-0.76), relapsers (risk ratio, 0.63; 95% CI, 0.54 0.73), and nonresponders (risk ratio, 0.89; 95% CI, 0.84-0.94). Combination therapy also significantly improved liver histologic response. The effects on quality of life are unclear. However, combination therapy significantly increased the risk of hematological, dermatological, gastrointestinal, and several other types of adverse events. In conclusion, the effect of ribavirin plus interferon on viral clearance may lead to reduced mortality and morbidity in patients with chronic hepatitis C infection. However, combination therapy is associated with increased risk for adverse events. PMID- 16246985 TI - A prospective study of anemia status, hemoglobin concentration, and mortality in an elderly cohort: the Cardiovascular Health Study. AB - BACKGROUND: Anemia is viewed as a negative prognostic factor in the elderly population; its independent impact on survival is unclear. METHODS: Baseline hemoglobin quintiles and anemia, as defined by the World Health Organization criteria, were assessed in relation to mortality in the Cardiovascular Health Study, a prospective cohort study with 11.2 years of follow-up of 5888 community dwelling men and women 65 years or older, enrolled in 1989-1990 or 1992-1993 in 4 US communities. RESULTS: A total of 1205 participants were in the lowest hemoglobin quintile (<13.7 g/dL for men; <12.6 g/dL for women), and 498 (8.5%) were anemic (<13 g/dL for men; <12 g/dL for women). A reverse J-shaped relationship with mortality was observed; age-, sex-, and race-adjusted hazard ratios (95% confidence interval [CI]) in the first and fifth quintiles, compared with the fourth quintile, were 1.42 (95% CI, 1.25-1.62) and 1.24 (95% CI, 1.09 1.42). After multivariate adjustment, these hazard ratios were 1.33 (95% CI, 1.15 1.54) and 1.17 (95% CI, 1.01-1.36). The demographic- and fully-adjusted hazard ratios of anemia for mortality were 1.57 (95% CI, 1.38-1.78) and 1.38 (95% CI, 1.19-1.54). Adjustment for causes and consequences of anemia (renal function, inflammation, or frailty) did not reduce associations. CONCLUSIONS: Lower and higher hemoglobin concentrations and anemia by World Health Organization criteria were independently associated with increased mortality. The World Health Organization criteria did not identify risk as well as a lower hemoglobin value. Additional study is needed on the clinically valid definition for and causes of anemia in the elderly and on the increased mortality at the extremes of hemoglobin concentrations. PMID- 16246987 TI - Renal function, erythropoietin, and anemia of older persons: the InCHIANTI study. AB - BACKGROUND: In the older population, anemia has been associated with poor outcomes including disability and mortality. Understanding the mechanisms leading to anemia is essential to plan better treatment and prevention strategies. We tested the hypothesis that the age-related decline in kidney function is associated with an increased prevalence of anemia and that such an increase is accompanied by a concomitant decrement in erythropoietin levels. METHODS: Data were from the InCHIANTI study, a population-based study performed in a sample of community-dwelling older (> or = 65 years) persons living in Italy. This analysis included 1005 participants with complete data on hemoglobin and erythropoietin levels and markers of renal function. RESULTS: The prevalence of anemia according to the World Health Organization criteria (hemoglobin level < 12 g/dL for women and < 13 g/dL for men) was 12.0% and increased with age in both sexes. After adjusting for age, diseases, and other confounders, only participants with a creatinine clearance (CrCl) of 30 mL/min or lower (< or = 0.50 mL/s) had a higher prevalence of anemia compared with those with a CrCl higher than 90 mL/min (> 1.50 mL/s) (P<.01). Consistently, participants with a CrCl of 30 mL/min or lower (< or = 0.50 mL/s) had significantly lower age- and hemoglobin-adjusted erythropoietin endogenous levels. After excluding men and women with CrCl of 30 mL/min or lower (< or = 0.50 mL/s) and adjusting for confounders, we found a trend toward an increase in prevalence of anemia with decreasing renal function; however, it was not statistically significant. CONCLUSIONS: Severe age-related decline in renal function is associated with a reduced erythropoietin secretion and anemia. Whether moderate kidney impairment in older persons is associated with a progressively increasing risk of anemia remains to be determined. PMID- 16246988 TI - The impact of anemia on energy and physical functioning in individuals with AIDS. AB - BACKGROUND: To our knowledge, the relationship between changes in hemoglobin level and energy and physical functioning in the anemic and "normal" ranges of hemoglobin among individuals with AIDS has not been well characterized. METHODS: In a multicenter, prospective, cohort study involving 19 clinics in the United States, 1406 individuals, 13 years and older, with AIDS were administered the Medical Outcomes Study HIV [human immunodeficiency virus] Health Survey (MOS-HIV) at baseline and at 3- to 6-month follow-up visits. Energy and physical functioning scores were the main outcomes. RESULTS: At baseline, a higher hemoglobin level was associated with a higher energy score and a higher physical functioning score (P < .001 for both), after adjusting for CD4 lymphocyte count, sex, age, education, and HIV risk factor. In longitudinal analyses, increases in hemoglobin were associated with increases in energy and physical functioning scores (P < .001 for both), after adjusting for CD4 lymphocyte count, sex, age, education, and HIV risk factor. Changes in the energy scales were, on average, 1.5 and 2.3 scale points per 1-g/dL change in hemoglobin level in the normal and anemic ranges, respectively. For the physical functioning scale, average changes were 2.7 and 2.6 scale points per 1-g/dL change in hemoglobin level in the normal and anemic ranges, respectively. CONCLUSIONS: Higher levels of hemoglobin are associated with better quality of life among individuals with AIDS. Changes in hemoglobin level within the conventional normal range of hemoglobin are also significantly associated with changes in quality of life. PMID- 16246989 TI - Anemia and outcomes in patients with heart failure: a study from the National Heart Care Project. AB - BACKGROUND: Recent reports have suggested that anemia is associated with adverse outcomes in patients with heart failure (HF), but were unable to adjust for a broad range of comorbid conditions. As a result, whether anemia is a truly independent predictor of risk or a marker of comorbid illness in these patients is unknown. METHODS: We analyzed medical records from the Centers for Medicare & Medicaid Services' National Heart Care Project, a national sample of 50,405 patients 65 years and older admitted to acute care hospitals with a principal discharge diagnosis of HF between April 1, 1998, and March 31, 1999, or between July 1, 2000, and June 30, 2001. Multivariable logistic regression analyses were conducted to test whether hematocrit level was an independent predictor of all cause mortality and HF-related readmission at 1 year. RESULTS: In unadjusted analysis, lower hematocrit levels were associated with increased 1-year mortality and readmission for HF. Compared with patients with a hematocrit greater than 40% to 44%, those with a hematocrit of 24% or less had a 51% higher risk of death (relative risk [RR], 1.51; 95% confidence interval [CI], 1.35-1.68; P<.001) and a 17% higher risk of HF-related readmission (RR, 1.17; 95% CI, 1.01-1.34; P = .04). However, after adjustment for multiple comorbidities and other clinical factors, the association between lower hematocrit levels and increased 1-year mortality was markedly attenuated, even in those patients with the most severe anemia (hematocrit, < or = 24% vs > 40%-44%: RR, 1.02; 95% CI, 0.86-1.19; P = .85). The association between lower hematocrit values and HF-related readmission persisted after multivariable adjustment (hematocrit, < or = 24% vs > 40%-44%: RR, 1.21; 95% CI, 1.04-1.38; P = .01). CONCLUSIONS: Although anemia is an independent predictor of hospital readmission, its relationship with increased mortality in HF patients is largely explained by the severity of comorbid illness. These findings suggest that anemia may be predominantly a marker rather than a mediator of increased mortality risk in older patients with HF. PMID- 16246990 TI - Justice at work and reduced risk of coronary heart disease among employees: the Whitehall II Study. AB - BACKGROUND: Justice is a fundamental value in human societies, but its effect on health is poorly described. We examined justice at work as a predictor of coronary heart disease (CHD). METHODS: Prospective occupational cohort study of 6442 male British civil servants aged 35 to 55 years without prevalent CHD at baseline in phase 1 (1985-1988). Baseline screening included measurements of conventional risk factors. Perceived justice at work and other work-related psychosocial factors were determined by means of questionnaire at phases 1 and 2 (1989-1990). Follow-up for CHD death, first nonfatal myocardial infarction, or definite angina occurring from phase 2 through 1999 was based on medical records (mean follow-up, 8.7 years). RESULTS: Cox proportional hazard models adjusted for age and employment grade showed that employees who experienced a high level of justice at work had a lower risk of incident CHD than employees with a low or an intermediate level of justice (hazard ratio, 0.65; 95% confidence interval, 0.47 0.89). The hazard ratio did not materially change after additional adjustment for baseline cholesterol concentration, body mass index, hypertension, smoking, alcohol consumption, and physical activity. Although other psychosocial models such as job strain and effort-reward imbalance predicted CHD in these data, the level of justice remained an independent predictor of incident CHD after adjustment for these factors. CONCLUSION: Justice at work may have benefits for heart health among employees. PMID- 16246991 TI - Variation in the tendency of primary care physicians to intervene. AB - BACKGROUND: Research has documented dramatic variation in health care spending across the United States that has little relationship to health outcomes. Although high-spending areas have more physicians per capita, it is not known whether this disparity fully explains the differences in spending or whether individual physicians in high-spending regions have a greater tendency to intervene for their patients. We sought to measure the tendency of primary care physicians to intervene across regions that differ in their levels of local health care spending. METHODS: We used data from the Community Tracking Study Physician Survey, a telephone survey of a nationally representative sample of 5490 primary care physicians who provided care to adults in 1998-1999 (response rate 59%). Local health care spending in physicians' communities was determined by assigning each participating physician to 1 of 306 US hospital referral regions. The tendency of physicians to intervene was measured by evaluating their responses to 6 clinical vignettes in which they were asked how often they would order a test, referral, or treatment for the patient described. RESULTS: In 5 of the 6 vignettes, physicians in high-spending regions were more likely to recommend interventions than those practicing in low-spending regions. For example, for a 35-year-old man with back pain and foot drop, physicians in high spending regions would recommend magnetic resonance imaging 82% of the time, compared with 69% for physicians in low-spending regions (P<.001). For a 60-year old man somewhat bothered by symptoms of benign prostatic hypertrophy, physicians in high-spending regions would make a urology referral 32% of the time, while those in low-spending regions would do so only 23% of the time (P<.001). Our findings that physicians in high-spending regions have a greater tendency to intervene persisted in analyses stratified by physician specialty (family/general practice vs internal medicine). CONCLUSION: Varying rates of health care spending across the United States reflect the underlying tendency of local physicians to recommend interventions for their patients. PMID- 16246992 TI - Initial patterns of clinical care and recovery from whiplash injuries: a population-based cohort study. AB - BACKGROUND: Little is known about the most effective pattern of clinical care for acute whiplash. We designed a cohort study to determine whether patterns of early clinical care (involving visits to general practitioners, chiropractors, or specialists) were associated with different rates of recovery. METHODS: We studied 2486 Saskatchewan adults with whiplash injuries. We defined 8 initial patterns of care that integrated type of provider and number of visits. We used multivariable Cox models to estimate the association between patterns of care and time to recovery while controlling for injury severity and other confounders. RESULTS: There was an independent association between the type and intensity of initial clinical care and time to recovery. We found that patients in the low utilization general practitioner group had the fastest recovery, even after controlling for injury severity and other confounders. Compared with this group, the high-utilization general practitioner group experienced a 1-year rate of recovery that was 27% slower (adjusted hazard rate ratio [HRR], 0.73; 95% confidence interval [CI], 0.61-0.87); for the high-utilization chiropractic group it was 39% slower (HRR, 0.61; 95% CI, 0.46-0.81); for the high-utilization general practitioner plus chiropractic combined group it was 28% slower (HRR, 0.72; 95% CI, 0.57-0.91); and for those who consulted general practitioners and specialists, it was 31% slower (HRR, 0.69; 95% CI, 0.55-0.87). CONCLUSIONS: The type and intensity of clinical care initiated within the first month after the injury is associated with the rate of recovery from whiplash injuries. Our study does not support the hypothesis that early aggressive care promotes faster recovery. PMID- 16246993 TI - Serum lipids, lipid-lowering drugs, and the risk of breast cancer. AB - BACKGROUND: Experimental evidence suggests that statins protect against breast carcinogenesis by interrupting cell cycle progression and promoting apoptosis. Evidence in humans is limited and inconsistent. The relation between serum cholesterol levels and breast cancer risk is itself unclear; because cholesterol is the precursor to sex steroid hormones, higher levels could plausibly increase risk. METHODS: The associations of statins, general lipid-lowering drugs, and reported cholesterol levels with breast cancer risk were assessed in the Nurses' Health Study, with 6 to 12 years of follow-up. A total of 79,994 women aged 42 to 69 years and free of cancer were followed prospectively for up to 12 years. Current statin use, including duration, was assessed retrospectively in 2000 in 75,828 women. Self-reported serum cholesterol level was assessed prospectively between 1990 and 2000 in 71,921 women. RESULTS: Overall, we documented 3177 incident cases of invasive breast cancer. Compared with nonusers, current lipid lowering drug users experienced similar breast cancer risk (multivariate relative risk [RR], 0.99; 95% confidence interval [CI], 0.86-1.13). Current use of statins also was not significantly associated with breast cancer risk (RR, 0.91; 95% CI, 0.76-1.08). Associations by duration of current use were similarly null. Self reported serum cholesterol levels were not associated with breast cancer risk in postmenopausal women with levels of 240 mg/dL or higher (> or = 6.22 mmol/L) compared with less than 180 mg/dL (< 4.66 mmol/L) (RR, 1.04; 95% CI, 0.91-1.17). CONCLUSION: Overall, these data suggest that serum cholesterol levels and the use of lipid-lowering drugs in general and of statins in particular are not substantially associated with breast cancer risk. PMID- 16246994 TI - Effect of a clinical trial alert system on physician participation in trial recruitment. AB - BACKGROUND: Failure to recruit a sufficient number of eligible subjects in a timely manner represents a major impediment to the success of clinical trials. Physician participation is vital to trial recruitment but is often limited. METHODS: After 12 months of traditional recruitment to a clinical trial, we activated our electronic health record (EHR)-based clinical trial alert (CTA) system in selected outpatient clinics of a large, US academic health care system. When a patient's EHR data met selected trial criteria during the subsequent 4 month intervention period, the CTA prompted physician consideration of the patient's eligibility and facilitated secure messaging to the trial's coordinator. Subjects were the 114 physicians practicing at selected EHR-equipped clinics throughout our study. We compared differences in the number of physicians participating in recruitment and their recruitment rates before and after CTA activation. RESULTS: The CTA intervention was associated with significant increases in the number of physicians generating referrals (5 before and 42 after; P < .001) and enrollments (5 before and 11 after; P = .03), a 10-fold increase in those physicians' referral rate (5.7/mo before and 59.5/mo after; rate ratio, 10.44; 95% confidence interval, 7.98-13.68; P<.001), and a doubling of their enrollment rate (2.9/mo before and 6.0/mo after; rate ratio, 2.06; 95% confidence interval, 1.22-3.46; P = .007). CONCLUSIONS: Use of an EHR-based CTA led to significant increases in physicians' participation in and recruitment rates to an ongoing clinical trial. Given the trend toward the EHR implementation in health care centers engaged in clinical research, this approach may represent a much-needed solution to the common problem of inadequate trial recruitment. PMID- 16246995 TI - Epidemiologic study of the autoimmune health effects of a cargo aircraft disaster. AB - BACKGROUND: In the aftermath of a cargo aircraft crash in Amsterdam in 1992, indications of autoimmune disorders appeared in some of the affected population. METHODS: This epidemiologic study sought to determine the possible long-term autoimmune health effects of the aircraft disaster on professional assistance workers. Exposed professional firefighters (n = 334) and police officers (n = 834) who performed at least 1 disaster-related task and hangar workers who sorted and investigated the wreckage (n = 241) were compared with reference groups of nonexposed colleagues who did not perform any disaster-related tasks (n = 194, n = 634, and n = 104, respectively). Data were collected a mean of 8.5 years after the disaster. Questionnaires were used to assess disaster-related tasks and 11 autoimmune-like symptoms. All serum samples were tested for the presence of antinuclear antibodies, rheumatoid factor, and antineutrophil cytoplasmic and anticardiolipin antibodies. RESULTS: Compared with nonexposed colleagues, exposed workers reported significantly more autoimmune-like symptoms. They reported the following symptoms significantly more often: tingling sensations, myalgia, loss of strength, easily fatigued, and a feeling of sand in the eyes (all groups); infection proneness (firefighters); skin abnormalities and nocturnal transpiration (police officers and hangar workers); and vasculitis-like symptoms and Raynaud discoloring (police officers). In contrast, we found no significant difference between exposed and nonexposed workers in autoantibody prevalence. CONCLUSION: Occupational exposure to the aircraft disaster resulted in an excess of long-term self-reported autoimmune-like symptoms in exposed professional assistance workers, but there was no difference between exposed and nonexposed workers in the prevalence of autoantibodies. PMID- 16246996 TI - Efficacy of bupropion and nortriptyline for smoking cessation among people at risk for or with chronic obstructive pulmonary disease. AB - BACKGROUND: The observations that smokers with chronic obstructive pulmonary disease (COPD) are at increased risk of depression and that nicotine may have antidepressant effects and regulate mood provide a rationale for the use of antidepressant drugs for smoking cessation in patients with COPD. No clinical trial has studied the efficacy of bupropion hydrochloride and nortriptyline hydrochloride for smoking cessation in this patient population, to our knowledge. METHODS: In a placebo-controlled double-dummy randomized trial, 255 adults at risk for COPD or with COPD were prescribed sustained-release bupropion (bupropion SR) (150 mg twice daily) or nortriptyline (75 mg once daily) for 12 weeks. All patients received smoking cessation counseling. The main outcome measure was prolonged abstinence from smoking from week 4 to week 26 after the target quit date. RESULTS: The use of bupropion SR and nortriptyline resulted in higher prolonged abstinence rates compared with placebo, although only the difference between bupropion SR and placebo was statistically significant (differences with placebo, 13.1% [95% confidence interval, 1.2%-25.1%] for bupropion SR and 10.2% [95% confidence interval, -1.7% to 22.2%] for nortriptyline). In patients with COPD, bupropion SR and nortriptyline seem efficacious in achieving prolonged abstinence (differences with placebo, 18.9% [95% confidence interval, 3.6%-34.2%] for bupropion SR and 12.9% [95% confidence interval, -0.8% to 26.4%] for nortriptyline). In participants at risk for COPD, no statistically significant differences with placebo in prolonged abstinence rates were found. CONCLUSIONS: Bupropion SR treatment is an efficacious aid to smoking cessation in patients with COPD. Nortriptyline treatment seems to be a useful alternative. PMID- 16246997 TI - Prevention of fall-related injuries in long-term care: a randomized controlled trial of staff education. AB - BACKGROUND: Fall-related injuries, a major public health problem in long-term care, may be reduced by interventions that improve safety practices. Previous studies have shown that safety practice interventions can reduce falls; however, in long-term care these have relied heavily on external funding and staff. The aim of this study was to test whether a training program in safety practices for staff could reduce fall-related injuries in long-term care facilities. METHODS: A cluster randomization clinical trial with 112 qualifying facilities and 10,558 study residents 65 years or older and not bedridden. The intervention was an intensive 2-day safety training program with 12-month follow-up. The training program targeted living space and personal safety; wheelchairs, canes, and walkers; psychotropic medication use; and transferring and ambulation. The main outcome measure was serious fall-related injuries during the follow-up period. RESULTS: There was no difference in injury occurrence between the intervention and control facilities (adjusted rate ratio, 0.98; 95% confidence interval, 0.83 1.16). For residents with a prior fall in facilities with the best program compliance, there was a nonsignificant trend toward fewer injuries in the intervention group (adjusted rate ratio, 0.79; 95% confidence interval, 0.57 1.10). CONCLUSION: More intensive interventions are required to prevent fall related injuries in long-term care facilities. PMID- 16246998 TI - Direct, progressive association of cardiovascular risk factors with incident proteinuria: results from the Korea Medical Insurance Corporation (KMIC) study. AB - BACKGROUND: Proteinuria is a major risk factor for the progression of kidney disease and the development of cardiovascular disease. Little is known, however, about risk factors for incident proteinuria. METHODS: We conducted a 10-year prospective cohort study of 104,523 Korean men and 52,854 women, aged 35 to 59 years, who attended Korea Medical Insurance Corporation health examinations and who did not have proteinuria at baseline. Incident proteinuria was assessed at biennial examinations during the next 10 years. We performed Cox proportional hazards analyses. RESULTS: During 10 years of follow-up, proteinuria developed in 3951 men (3.8%) and 1527 women (2.9%). The adjusted relative risk (RR) of proteinuria associated with diabetes was 3.27 (95% confidence interval [CI], 2.98 3.58) in men and 2.60 (95% CI, 1.98-3.43) in women; with body mass index (calculated as weight in kilograms divided by the square of height in meters), it was 1.43 (95% CI, 1.35-1.50) in men and 1.45 (95% CI, 1.35-1.55) in women per 5-U increment. Compared with subjects with serum cholesterol levels of less than 200 mg/dL (< 5.18 mmol/L), the adjusted RRs associated with serum cholesterol levels of 200 to 239 mg/dL (5.18-6.19 mmol/L) and 240 mg/dL or more (> or = 6.22 mmol/L) were 1.13 (95% CI, 1.05-1.21) and 1.40 (95% CI, 1.27-1.54), respectively, in men and 1.14 (95% CI, 1.01-1.28) and 1.22 (95% CI, 1.00-1.37), respectively, in women. Persons with stages 1 and 2 hypertension had a greater adjusted RR of incident proteinuria compared with those with normal blood pressure (1.62 [95% CI, 1.47-1.79] and 2.06 [95% CI, 1.81-2.34], respectively, in men and 1.37 [95% CI, 1.14-1.65] and 2.10 [95% CI, 1.59-2.76], respectively, in women). CONCLUSIONS: Fasting glucose and cholesterol levels, body mass index, and blood pressure were direct and independent predictors of incident proteinuria in Korean adults. These associations were present even at low levels of exposure, emphasizing the importance of early detection and management of these modifiable risk factors. PMID- 16246999 TI - 3D structure of eukaryotic flagella in a quiescent state revealed by cryo electron tomography. AB - We have used cryo-electron tomography to investigate the 3D structure and macromolecular organization of intact, frozen-hydrated sea urchin sperm flagella in a quiescent state. The tomographic reconstructions provide information at a resolution better than 6 nm about the in situ arrangements of macromolecules that are key for flagellar motility. We have visualized the heptameric rings of the motor domains in the outer dynein arm complex and determined that they lie parallel to the plane that contains the axes of neighboring flagellar microtubules. Both the material associated with the central pair of microtubules and the radial spokes display a plane of symmetry that helps to explain the planar beat pattern of these flagella. Cryo-electron tomography has proven to be a powerful technique for helping us understand the relationships between flagellar structure and function and the design of macromolecular machines in situ. PMID- 16247000 TI - Evidence for maternally transmitted small interfering RNA in the repression of transposition in Drosophila virilis. AB - Hybrid dysgenesis in Drosophila is a syndrome of gonadal atrophy, sterility, and male recombination, and it occurs in the progeny of crosses between males that harbor certain transposable elements (TEs) and females that lack them. Known examples of hybrid dysgenesis in Drosophila melanogaster result from mobilization of individual families of TEs, such as the P element, the I element, or hobo. An example of hybrid dysgenesis in Drosophila virilis is unique in that multiple, unrelated families of TEs become mobilized, but a TE designated Penelope appears to play a major role. In all known examples of hybrid dysgenesis, the paternal germ line transmits the TEs in an active state, whereas the female germ line maintains repression of the TEs. The mechanism of maternal maintenance of repression is not known. Recent evidence suggests that the molecular machinery of RNA interference may function as an important host defense against TEs. This protection is mediated by the action of endogenous small interfering RNAs (siRNAs) composed of dsRNA molecules of 21-25 nt that can target complementary transcripts for destruction. In this paper, we demonstrate that endogenous siRNA derived from the Penelope element is maternally loaded in embryos through the female germ line in D. virilis. We also present evidence that the maternal inheritance of these endogenous siRNAs may contribute to maternal repression of Penelope. PMID- 16247001 TI - The role of TNF-alpha in the pathogenesis of type 1 diabetes in the nonobese diabetic mouse: analysis of dendritic cell maturation. AB - TNF-alpha has been linked to the development of type 1 diabetes (T1D). We previously reported that neonatal treatment of nonobese diabetic (NOD) mice with TNF-alpha accelerated the onset of T1D, whereas TNF-alpha blockade in the same time period resulted in a complete absence of diabetes. The mechanisms by which TNF-alpha modulates development of T1D in NOD mice remain unclear. Here we tested the effects of TNF-alpha on the maturation of dendritic cells (DCs) in the NOD mouse. We found that neonatal treatment with TNF-alpha caused an increase in expression of maturation markers on CD11c(+)CD11b(+) DC subpopulations, whereas treatment with anti-TNF-alpha resulted in a decrease in expression of maturation markers in the CD11c(+)CD11b(+) subset. Moreover, neonatal treatment with TNF alpha resulted in skewed development of a CD8alpha(+)CD11b(-)CD11c(+) DC subset such that TNF-alpha decreases the CD8alpha(+)CD11c(+) DC subset, increases the CD11c(+)CD11b(+) subset, and causes an increase in the expression of CD40 and CD54 on mature DCs capable of inducing immunity. Anti-TNF-alpha-treated mice had an increase in the CD8alpha(+)CD11c(+) DCs. Notably, adoptively transferred naive CD4(+) T cells from BDC2.5 T cell receptor transgenic mice proliferated in the pancreatic lymph nodes in TNF-alpha-treated NOD mice but not in anti-TNF-alpha treated mice. Finally, we show that anti-TNF-alpha-treated mice showed immunological tolerance to islet cell proteins. We conclude that TNF-alpha plays an important role in the initiation of T1D in the NOD mouse by regulating the maturation of DCs and, thus, the activation of islet-specific pancreatic lymph node T cells. PMID- 16247002 TI - The sodium pump and hypertension: a physiological role for the cardiac glycoside binding site of the Na,K-ATPase. PMID- 16247003 TI - Diapause in the mosquito Culex pipiens evokes a metabolic switch from blood feeding to sugar gluttony. AB - A key characteristic of overwintering dormancy (diapause) in the mosquito Culex pipiens is the switch in females from blood feeding to sugar gluttony. We present evidence demonstrating that genes encoding enzymes needed to digest a blood meal (trypsin and a chymotrypsin-like protease) are down-regulated in diapause destined females, and that concurrently, a gene associated with the accumulation of lipid reserves (fatty acid synthase) is highly up-regulated. As the females then enter diapause, fatty acid synthase is only sporadically expressed, and expression of trypsin and chymotrypsin-like remains undetectable. Late in diapause (2-3 months at 18 degrees C), the genes encoding the digestive enzymes begin to be expressed as the female prepares to take a blood meal upon the termination of diapause. Our results thus underscore a molecular switch that either capacitates the mosquito for blood feeding (nondiapause) or channels the adult mosquito exclusively toward sugar feeding and lipid sequestration (diapause). PMID- 16247004 TI - Structure of the metal-independent restriction enzyme BfiI reveals fusion of a specific DNA-binding domain with a nonspecific nuclease. AB - Among all restriction endonucleases known to date, BfiI is unique in cleaving DNA in the absence of metal ions. BfiI represents a different evolutionary lineage of restriction enzymes, as shown by its crystal structure at 1.9-A resolution. The protein consists of two structural domains. The N-terminal catalytic domain is similar to Nuc, an EDTA-resistant nuclease from the phospholipase D superfamily. The C-terminal DNA-binding domain of BfiI exhibits a beta-barrel-like structure very similar to the effector DNA-binding domain of the Mg(2+)-dependent restriction enzyme EcoRII and to the B3-like DNA-binding domain of plant transcription factors. BfiI presumably evolved through domain fusion of a DNA recognition element to a nonspecific nuclease akin to Nuc and elaborated a mechanism to limit DNA cleavage to a single double-strand break near the specific recognition sequence. The crystal structure suggests that the interdomain linker may act as an autoinhibitor controlling BfiI catalytic activity in the absence of a specific DNA sequence. A psi-blast search identified a BfiI homologue in a Mesorhizobium sp. BNC1 bacteria strain, a plant symbiont isolated from an EDTA rich environment. PMID- 16247005 TI - Involvement of winged eye encoding a chromatin-associated bromo-adjacent homology domain protein in disc specification. AB - How organ identity is determined is a fundamental question in developmental biology. In Drosophila, field-specific selector genes, such as eyeless (ey) for eyes and vestigial (vg) for wings, participate in the determination of imaginal disc-specific identity. We performed gain-of-function screening and identified a gene named winged eye (wge), which encodes a bromo-adjacent homology domain protein that localizes at specific sites on chromosomes in a bromo-adjacent homology domain-dependent manner. Overexpression of wge-induced ectopic wings with antero-posterior and dorso-ventral axes in the eye field in a region specific Hox gene-(Antennapedia) independent manner. Overexpression of wge was sufficient for ectopic expression of vg in eye discs. A context-dependent requirement of wge was demonstrated for vg expression in wing discs and for expression of eyes absent (eya), a control gene for eye development downstream of ey, in eye discs. In contrast to vg, however, overexpression of wge inhibited EY mediated expression of eya. Consistent with colocalization on polytene chromosomes of WGE and Posterior sex combs (PSC), a Polycomb group gene product, we demonstrated an antagonistic genetic interaction between wge and Psc. These findings suggest that wge functions in the determination of disc-specific identity, downstream of Hox genes. PMID- 16247006 TI - Pore conformations and gating mechanism of a Cys-loop receptor. AB - Neurons regulate the propagation of chemoelectric signals throughout the nervous system by opening and closing ion channels, a process known as gating. Here, histidine-based metal-binding sites were engineered along the intrinsic pore of a chimeric Cys-loop receptor to probe state-dependent Zn(2+)-channel interactions. Patterns of Zn(2+) ion binding within the pore reveal that, in the closed state, the five pore-lining segments adopt an oblique orientation relative to the axis of ion conduction and constrict into a physical gate at their intracellular end. The interactions of Zn(2+) with the open state indicate that the five pore-lining segments should rigidly tilt to enable the movement of their intracellular ends away from the axis of ion conduction, so as to open the constriction (i.e., the gate). Alignment of the functional results with the 3D structure of an acetylcholine receptor allowed us to generate structural models accounting for the closed and open pore conformations and for a gating mechanism of a Cys-loop receptor. PMID- 16247007 TI - Breath sulfides and pulmonary function in cystic fibrosis. AB - We have determined the concentrations of carbonyl sulfide (OCS), dimethylsulfide, and carbon disulfide (CS(2)) in the breath of a group of cystic fibrosis (CF) patients and one of healthy controls. At the detection sensitivity in these experiments, room air always contained measurable quantities of these three gases. For each subject the inhaled room concentrations were subtracted from the time-coincident concentrations in exhaled breath air. The most significant differences between the CF and control cohorts in these breath-minus-room values were found for OCS. The control group demonstrated a net uptake of 250 +/- 20 parts-per-trillion-by-volume (pptv), whereas the CF cohort had a net uptake of 110 +/- 60 pptv (P = 0.00003). Three CF patients exhaled more OCS than they inhaled from the room. The OCS concentrations in the CF cohort were strongly correlated with pulmonary function. The dimethylsulfide concentrations in breath were greatly enhanced over ambient, but no significant difference was observed between the CF and healthy control groups. The net (breath minus room) CS(2) concentrations for individuals ranged between +180 and -100 pptv. They were slightly greater in the CF cohort (+26 +/- 38 pptv) vs. the control group (-17 +/ 15 pptv; P = 0.04). Lung disease in CF is accompanied by the subsistence of chronic bacterial infections. Sulfides are known to be produced by bacteria in various systems and were therefore the special target for this investigation. Our results suggest that breath sulfide content deserves attention as a noninvasive marker of respiratory colonization. PMID- 16247008 TI - Molecular-level secondary structure, polymorphism, and dynamics of full-length alpha-synuclein fibrils studied by solid-state NMR. AB - The 140-residue protein alpha-synuclein (AS) is able to form amyloid fibrils and as such is the main component of protein inclusions involved in Parkinson's disease. We have investigated the structure and dynamics of full-length AS fibrils by high-resolution solid-state NMR spectroscopy. Homonuclear and heteronuclear 2D and 3D spectra of fibrils grown from uniformly (13)C/(15)N labeled AS and AS reverse-labeled for two of the most abundant amino acids, K and V, were analyzed. (13)C and (15)N signals exhibited linewidths of <0.7 ppm. Sequential assignments were obtained for 48 residues in the hydrophobic core region. We identified two different types of fibrils displaying chemical-shift differences of up to 13 ppm in the (15)N dimension and up to 5 ppm for backbone and side-chain (13)C chemical shifts. EM studies suggested that molecular structure is correlated with fibril morphology. Investigation of the secondary structure revealed that most amino acids of the core region belong to beta strands with similar torsion angles in both conformations. Selection of regions with different mobility indicated the existence of monomers in the sample and allowed the identification of mobile segments of the protein within the fibril in the presence of monomeric protein. At least 35 C-terminal residues were mobile and lacked a defined secondary structure, whereas the N terminus was rigid starting from residue 22. Our findings agree well with the overall picture obtained with other methods and provide insight into the amyloid fibril structure and dynamics with residue-specific resolution. PMID- 16247009 TI - Cooperative effects of Rho and mechanical stretch on stress fiber organization. AB - The small GTPase Rho regulates the formation of actin stress fibers in adherent cells through activation of its effector proteins Rho-kinase and mDia. We found in bovine aortic endothelial cells that inhibitions of Rho, Rho-kinase, and mDia (with C3, Y27632, and F1F2Delta1, respectively) suppressed stress fiber formation, but fibers appeared after 10% cyclic uniaxial stretch (1-Hz frequency). In contrast to the predominately perpendicular alignment of stress fibers to the stretch direction in normal cells, the stress fibers in cells with Rho pathway inhibition became oriented parallel to the stretch direction. In cells with normal Rho activity, the extent of perpendicular orientation of stress fibers depended on the magnitude of stretch. Expressing active RhoV14 plasmid in these cells enhanced the stretch-induced stress fiber orientation by an extent equivalent to an additional approximately 3% stretch. This augmentation of the stretch-induced perpendicular orientation by RhoV14 was blocked by Y27632 and by F1F2Delta1. Thus, the activity of the Rho pathway plays a critical role in determining both the direction and extent of stretch-induced stress fiber orientation in bovine aortic endothelial cells. Our results demonstrate that the stretch-induced stress fiber orientation is a function of the interplay between Rho pathway activity and the magnitude of stretching. PMID- 16247010 TI - Haploinsufficiency of telomerase reverse transcriptase leads to anticipation in autosomal dominant dyskeratosis congenita. AB - Dyskeratosis congenita is a rare inherited disorder characterized by abnormal skin manifestations. Morbidity and mortality from this disease is usually due to bone marrow failure, but idiopathic pulmonary fibrosis and an increased cancer predisposition also occur. Families with autosomal dominant dyskeratosis congenita display anticipation and have mutations in the telomerase RNA gene. We identified a three-generation pedigree with autosomal dominant dyskeratosis congenita, anticipation, and telomere shortening. We show that a null mutation in motif D of the reverse transcriptase domain of the protein component of telomerase, hTERT, is associated with this phenotype. This mutation leads to haploinsufficiency of telomerase, and telomere shortening occurs despite the presence of telomerase. This finding emphasizes the importance of telomere maintenance and telomerase dosage for maintaining tissue proliferative capacity and has relevance for understanding mechanisms of age-related changes. PMID- 16247011 TI - Herpesviral latency-associated transcript gene promotes assembly of heterochromatin on viral lytic-gene promoters in latent infection. AB - Herpes simplex virus (HSV) persists in its human host and evades the immune response by undergoing a latent infection in sensory neurons, from which it can reactivate periodically. HSV expresses >80 gene products during productive ("lytic") infection, but only the latency-associated transcript (LAT) gene is expressed at abundant levels during latent infection. The LAT gene has been shown to repress lytic-gene expression in sensory neurons. In this study, we use chromatin immunoprecipitation to show that HSV lytic-gene promoters become complexed with modified histones associated with heterochromatin during the course of establishment of latent infection. Experiments comparing LAT-negative and LAT-positive viruses show that a function encoded by the LAT gene increases the amount of dimethyl lysine 9 form of histone H3 or heterochromatin and reduces the amount of dimethyl lysine 4 form of histone H3, a part of active chromatin, on viral lytic-gene promoters. Thus, HSV, and in particular the HSV LAT gene, may manipulate the cellular histone modification machinery to repress its lytic-gene expression and contribute to the persistence of its genome in a quiescent form in sensory neurons. PMID- 16247012 TI - A structural model for maturation of the hepatitis B virus core. AB - Hepatitis B virus, a widespread and serious human pathogen, replicates by reverse transcription of an RNA intermediate. The virus consists of an inner nucleocapsid or core, surrounded by a lipid envelope containing virally encoded surface proteins. Using electron cryomicroscopy, we compare the structures of the bacterially expressed RNA-containing core particle and the mature DNA-containing core particle extracted from virions. We show that the mature core contains 240 subunits in a T = 4 arrangement similar to that in expressed core (T is the triangulation number and the icosahedral shell contains 60 T subunits). During the infective cycle, the core assembles in an immature state around a complex of viral pregenomic RNA and polymerase. After reverse transcription with concomitant degradation of the RNA, the now mature core buds through a cellular membrane containing the surface proteins to become enveloped. Envelopment must not happen before reverse transcription is completed, so it has been hypothesized that a change in capsid structure may signal maturation. Our results show significant differences in structure between the RNA- and DNA-containing cores. One such difference is in a hydrophobic pocket, formed largely from residues that, on mutation, lead to abnormal secretion. We suggest that the changes we see are related to maturation and control of envelopment, and we propose a mechanism based on DNA synthesis for their triggering. PMID- 16247013 TI - Orbitofrontal cortex activity related to emotional processing changes across the menstrual cycle. AB - The orbitofrontal cortex (OFC) has been implicated in the representation of emotional stimuli, assignment of emotional valence/salience to stimuli, stimulus reinforcement association learning, motivation, and socio-emotional control. Using functional magnetic resonance imaging in female subjects without premenstrual mood symptoms, we found that OFC activity to emotional linguistic stimuli varies depending on the menstrual cycle phase. Specifically, anterior medial OFC activity for negative vs. neutral stimuli was increased premenstrually and decreased postmenstrually. The inverse pattern was seen in the lateral OFC. These findings suggest that specific subregional OFC activity to emotional stimuli is modulated across the menstrual cycle. The data also demonstrate that menstrual cycle phase is an important consideration in further studies attempting to elucidate the neural substrates of affective representation. PMID- 16247014 TI - The response of autologous T cells to a human melanoma is dominated by mutated neoantigens. AB - Our understanding of pathways leading to antitumor immunity may depend on an undistorted knowledge of the primary antigenic targets of patients' autologous T cell responses. In the melanoma model derived from patient DT, we applied cryopreserved short-term autologous mixed lymphocyte-tumor cell cultures (MLTCs) in combination with an IFN-gamma enzyme-linked immunospot (ELISPOT) assay to cDNA expression screening. We identified three previously unknown peptides processed from melanosomal proteins tyrosinase (presented by HLA-A(*)2601 and -B(*)3801) and gp100 (presented by HLA-B(*)07021) and five neoantigens generated by somatic point mutations in the patient's melanoma. The mutations were found in the genes SIRT2, GPNMB, SNRP116, SNRPD1, and RBAF600. Peptides containing the mutated residues were presented by HLA-A(*)03011, -B(*)07021, and -B(*)3801. Mutation induced functional impairment was so far demonstrated for SIRT2. Within MLTC responder populations that were independently expanded from the patient's peripheral blood lymphocytes of different years, T cells against mutated epitopes clearly predominated. These results document a high degree of individuality for the cellular antitumor response and support the need for individualizing the monitoring and therapeutic approaches to the primary targets of the autologous T cell response, which may finally lead to a more effective cancer immunotherapy. PMID- 16247015 TI - Metastatic properties and genomic amplification of the tyrosine kinase gene ACK1. AB - Metastasis of primary tumors leads to a very poor prognosis for patients suffering from cancer. Although it is well established that not every tumor will eventually metastasize, it is less clear whether primary tumors acquire genetic alterations in a stochastic process at a late stage, which make them invasive, or whether genetic alterations acquired early in the process of tumor development drive primary tumor growth and determine whether this tumor is going to be metastatic. To address this issue, we tested genes identified in a large-scale comparative genomic hybridization analysis of primary tumor for their ability to confer metastatic properties on a cancer cell. We identified amplification of the ACK1 gene in primary tumors, which correlates with poor prognosis. We further show that overexpression of Ack1 in cancer cell lines can increase the invasive phenotype of these cells both in vitro and in vivo and leads to increased mortality in a mouse model of metastasis. Biochemical studies show that Ack1 is involved in extracellular matrix-induced integrin signaling, ultimately activating signaling processes like the activation of the small GTPase Rac. Taken together, this study supports a theory from Bernards and Weinberg [Bernards, R. & Weinberg, R. A. (2002) Nature 418, 823], which postulates that the tendency to metastasize is largely predetermined. PMID- 16247016 TI - Prostaglandin E receptor EP1 controls impulsive behavior under stress. AB - Animals under stress take adaptive actions that may lead to various types of behavioral disinhibition. Such behavioral disinhibition, when expressed excessively and impulsively, can result in harm in individuals and cause a problem in our society. We now show that, under social or environmental stress, mice deficient in prostaglandin E receptor subtype EP1 (Ptger1(-/-)) manifest behavioral disinhibition, including impulsive aggression with defective social interaction, impaired cliff avoidance, and an exaggerated acoustic startle response. This phenotype was reproduced in wild-type mice by administration of an EP1-selective antagonist, whereas administration of an EP1-selective agonist suppressed electric-shock-induced impulsive aggression. Dopamine turnover in the frontal cortex and striatum was increased in Ptger1(-/-) mice, and administration of dopaminergic antagonists corrected their behavioral phenotype. These results suggest that prostaglandin E(2) acts through EP1 to control impulsive behavior under stress, a finding potentially exploitable for development of drugs that attenuate impulsive behavior in humans. PMID- 16247017 TI - The error-free component of the RAD6/RAD18 DNA damage tolerance pathway of budding yeast employs sister-strand recombination. AB - Evidence for an error-free DNA damage tolerance process in eukaryotes (also called postreplication repair) has existed for more than two decades, but its underlying mechanism, although known to be different from that in prokaryotes, has remained elusive. We have investigated this mechanism in Saccharomyces cerevisiae, in which it is the major component of the RAD6/RAD18 pathway, by transforming an isogenic set of rad1Delta excision-defective strains with plasmids that carry a single thymine-thymine pyrimidine (6-4) pyrimidinone photoadduct in each strand at staggered positions 28 base pairs apart. C-C mismatches placed opposite each of the T-T photoproducts permit unambiguous detection of the events that can lead to the completion of replication: sister strand recombination or translesion replication on one or the other strand. Despite the severe block to replication that these lesions impose, we find that more than half of the plasmids were fully replicated in a rad1Delta strain and that >90% of them achieved this end by recombination between partially replicated sister strands within the interlesion region. Approximately 60-70% of these events depended on the error-free component of the RAD6/RAD18 pathway, with the remaining events depended on RAD52; these two processes account for almost all of the recombination, which depended neither on DNA polymerase zeta nor on mismatch repair. We conclude that the error-free component of the RAD6/RAD18 pathway completes replication by a mechanism employing recombination between partially replicated sister strands, possibly by means of transient template strand switching or copy choice. PMID- 16247018 TI - Spectral and redox characterization of the heme ci of the cytochrome b6f complex. AB - Absorption spectra of the purified cytochrome b(6)f complex from Chlamydomonas reinhardtii were monitored as a function of the redox potential. Four spectral and redox components were identified: in addition to heme f and the two b hemes, the fourth component must be the new heme c(i) (also denoted x) recently discovered in the crystallographic structures. This heme is covalently attached to the protein, but has no amino acid axial ligand. It is located in the plastoquinone-reducing site Q(i) in the immediate vicinity of a b heme. Each heme titrated as a one-electron Nernst curve, with midpoint potentials at pH 7.0 of 130 mV and -35 mV (hemes b), +100 mV (heme c(i)), and +355 mV (heme f). The reduced minus oxidized spectrum of heme c(i) consists of a broad absorption increase centered approximately 425 nm. Its potential has a dependence of -60 mV/pH unit, implying that the reduced form binds one proton in the pH 6-9 range. The Q(i) site inhibitor 2-n-nonyl-4-hydroxyquinoline N-oxide, a semiquinone analogue, induces a shift of this potential by about -225 mV. The spectrum of c(i) matches the absorption changes previously observed in vivo for an unknown redox center denoted "G." The data are discussed with respect to the effect of the membrane potential on the electron transfer equilibrium between G and heme b(H) found in earlier experiments. PMID- 16247020 TI - Cofilin expression induces cofilin-actin rod formation and disrupts synaptic structure and function in Aplysia synapses. AB - Cofilin-actin rods are inclusion-like structures that are induced by certain chemical or physical stresses in cultured cells, and the rods formed in neurons are thought to be associated with neurodegeneration. Here, we cloned an Aplysia cofilin homolog and overexpressed it in cultured neurons. Overexpressed cofilin formed rod-like structures that included actin. The overall neuronal morphology was unaffected by cofilin overexpression; however, a decrease in number of synaptic varicosities was observed. Consistent with this structural change by cofilin overexpression, the synaptic strength was reduced, and furthermore, the long-term facilitation elicited by repeated pulses of 5-hydroxytryptamine was impaired in sensory-to-motor synapses. However, cofilin overexpression did not induce programmed cell death. These findings suggest that the formation of cofilin-actin rod-like structures can lead to neurodegeneration, and this might be a mechanism of rundown of neuronal and synaptic function without cell death in neurodegenerative diseases. PMID- 16247021 TI - TIP maker and TIP marker; EB1 as a master controller of microtubule plus ends. AB - The EB1 protein is a member of the exciting and enigmatic family of microtubule (MT) tip-tracking proteins. EB1 acts as an exquisite marker of dynamic MT plus ends in some cases, whereas in others EB1 is thought to directly dictate the behavior of the plus ends. How EB1 differentiates between these two roles remains unclear; however, a growing list of interactions between EB1 and other MT binding proteins suggests there may be a single mechanism. Adding another layer of complexity to these interactions, two studies published in this issue implicate EB1 in cross-talk between mitotic MTs and between MTs and actin filaments (Goshima et al., p. 229; Wu et al., p. 201). These results raise the possibility that EB1 is a central player in MT-based transport, and that the activity of MT binding proteins depends on their ability or inability to interact with EB1. PMID- 16247022 TI - Melanophilin and myosin Va track the microtubule plus end on EB1. AB - In mouse melanocytes, myosin Va is recruited onto the surface of melanosomes by a receptor complex containing Rab27a that is present in the melanosome membrane and melanophilin (Mlp), which links myosin Va to Rab27a. In this study, we show that Mlp is also a microtubule plus end-tracking protein or +TIP. Moreover, myosin Va tracks the plus end in a Mlp-dependent manner. Data showing that overexpression and short inhibitory RNA knockdown of the +TIP EB1 have opposite effects on Mlp microtubule interaction, that Mlp interacts directly with EB1, and that deletion from Mlp of a region similar to one in the adenomatous polyposis coli protein involved in EB1 binding blocks Mlp's ability to plus end track argue that Mlp tracks the plus end indirectly [corrected] by hitchhiking on EB1. These results identify a novel +TIP and indicate that vertebrate cells possess a +TIP complex that is similar to the Myo2p-Kar9p-Bim1p complex in yeast. We suggest that the +TIP complex identified in this study may serve to focus the transfer of melanosomes from microtubules to actin at the microtubule plus end. PMID- 16247023 TI - Mechanical force mobilizes zyxin from focal adhesions to actin filaments and regulates cytoskeletal reinforcement. AB - Organs and tissues adapt to acute or chronic mechanical stress by remodeling their actin cytoskeletons. Cells that are stimulated by cyclic stretch or shear stress in vitro undergo bimodal cytoskeletal responses that include rapid reinforcement and gradual reorientation of actin stress fibers; however, the mechanism by which cells respond to mechanical cues has been obscure. We report that the application of either unidirectional cyclic stretch or shear stress to cells results in robust mobilization of zyxin from focal adhesions to actin filaments, whereas many other focal adhesion proteins and zyxin family members remain at focal adhesions. Mechanical stress also induces the rapid zyxin dependent mobilization of vasodilator-stimulated phosphoprotein from focal adhesions to actin filaments. Thickening of actin stress fibers reflects a cellular adaptation to mechanical stress; this cytoskeletal reinforcement coincides with zyxin mobilization and is abrogated in zyxin-null cells. Our findings identify zyxin as a mechanosensitive protein and provide mechanistic insight into how cells respond to mechanical cues. PMID- 16247024 TI - Analysis of the Xenopus Werner syndrome protein in DNA double-strand break repair. AB - Werner syndrome is associated with premature aging and increased risk of cancer. Werner syndrome protein (WRN) is a RecQ-type DNA helicase, which seems to participate in DNA replication, double-strand break (DSB) repair, and telomere maintenance; however, its exact function remains elusive. Using Xenopus egg extracts as the model system, we found that Xenopus WRN (xWRN) is recruited to discrete foci upon induction of DSBs. Depletion of xWRN has no significant effect on nonhomologous end-joining of DSB ends, but it causes a significant reduction in the homology-dependent single-strand annealing DSB repair pathway. These results provide the first direct biochemical evidence that links WRN to a specific DSB repair pathway. The assay for single-strand annealing that was developed in this study also provides a powerful biochemical system for mechanistic analysis of homology-dependent DSB repair. PMID- 16247019 TI - Centromere-proximal differentiation and speciation in Anopheles gambiae. AB - The M and S molecular forms of Anopheles gambiae are undergoing speciation as they adapt to heterogeneities in the environment, spreading malaria in the process. We hypothesized that their divergence despite gene flow is facilitated by reduced recombination at the centromeric (proximal) end of the X chromosome. We sequenced introns from 22 X chromosome genes in M and S from two locations of West Africa where the forms are sympatric. Generally, in both forms nucleotide diversity was high distally, lower proximally, and very low nearest the centromere. Conversely, differentiation between the forms was virtually zero distally and very high proximally. Pairwise comparisons to a close relative, the sibling species Anopheles arabiensis, demonstrated uniformly high divergence regardless of position along the X chromosome, suggesting that this pattern is not purely mechanical. Instead, the pattern observed for M and S suggests the action of divergent natural selection countering gene flow only at the proximal end of the X chromosome, where recombination is reduced. Comparison of sites with fixed differences between M and S to the corresponding sites in A. arabiensis revealed that derived substitutions had been fixed in both forms, further supporting the hypothesis that both have been under selection. These derived substitutions are fixed in the two West African samples and in samples of S from western and coastal Kenya, suggesting that selection occurred before the forms expanded to their current ranges. Our findings are consistent with a role for suppressed genetic recombination in speciation of A. gambiae. PMID- 16247025 TI - Mechanisms for focusing mitotic spindle poles by minus end-directed motor proteins. AB - During the formation of the metaphase spindle in animal somatic cells, kinetochore microtubule bundles (K fibers) are often disconnected from centrosomes, because they are released from centrosomes or directly generated from chromosomes. To create the tightly focused, diamond-shaped appearance of the bipolar spindle, K fibers need to be interconnected with centrosomal microtubules (C-MTs) by minus end-directed motor proteins. Here, we have characterized the roles of two minus end-directed motors, dynein and Ncd, in such processes in Drosophila S2 cells using RNA interference and high resolution microscopy. Even though these two motors have overlapping functions, we show that Ncd is primarily responsible for focusing K fibers, whereas dynein has a dominant function in transporting K fibers to the centrosomes. We also report a novel localization of Ncd to the growing tips of C-MTs, which we show is mediated by the plus end tracking protein, EB1. Computer modeling of the K fiber focusing process suggests that the plus end localization of Ncd could facilitate the capture and transport of K fibers along C-MTs. From these results and simulations, we propose a model on how two minus end-directed motors cooperate to ensure spindle pole coalescence during mitosis. PMID- 16247026 TI - Regulation of meiotic prophase arrest in mouse oocytes by GPR3, a constitutive activator of the Gs G protein. AB - The arrest of meiotic prophase in mouse oocytes within antral follicles requires the G protein G(s) and an orphan member of the G protein-coupled receptor family, GPR3. To determine whether GPR3 activates G(s), the localization of Galpha(s) in follicle-enclosed oocytes from Gpr3(+/+) and Gpr3(-/-) mice was compared by using immunofluorescence and Galpha(s)GFP. GPR3 decreased the ratio of Galpha(s) in the oocyte plasma membrane versus the cytoplasm and also decreased the amount of Galpha(s) in the oocyte. Both of these properties indicate that GPR3 activates G(s). The follicle cells around the oocyte are also necessary to keep the oocyte in prophase, suggesting that they might activate GPR3. However, GPR3-dependent G(s) activity was similar in follicle-enclosed and follicle-free oocytes. Thus, the maintenance of prophase arrest depends on the constitutive activity of GPR3 in the oocyte, and the follicle cell signal acts by a means other than increasing GPR3 activity. PMID- 16247027 TI - A complex containing the Sm protein CAR-1 and the RNA helicase CGH-1 is required for embryonic cytokinesis in Caenorhabditis elegans. AB - Cytokinesis completes cell division and partitions the contents of one cell to the two daughter cells. Here we characterize CAR-1, a predicted RNA binding protein that is implicated in cytokinesis. CAR-1 localizes to germline-specific RNA-containing particles and copurifies with the essential RNA helicase, CGH-1, in an RNA-dependent fashion. The atypical Sm domain of CAR-1, which directly binds RNA, is dispensable for CAR-1 localization, but is critical for its function. Inhibition of CAR-1 by RNA-mediated depletion or mutation results in a specific defect in embryonic cytokinesis. This cytokinesis failure likely results from an anaphase spindle defect in which interzonal microtubule bundles that recruit Aurora B kinase and the kinesin, ZEN-4, fail to form between the separating chromosomes. Depletion of CGH-1 results in sterility, but partially depleted worms produce embryos that exhibit the CAR-1-depletion phenotype. Cumulatively, our results suggest that CAR-1 functions with CGH-1 to regulate a specific set of maternally loaded RNAs that is required for anaphase spindle structure and cytokinesis. PMID- 16247028 TI - The role of Fis1p-Mdv1p interactions in mitochondrial fission complex assembly. AB - Mitochondrial division requires coordinated interactions among Fis1p, Mdv1p, and the Dnm1p GTPase, which assemble into fission complexes on the outer mitochondrial membrane. The integral outer membrane protein Fis1p contains a cytoplasmic domain consisting of a tetratricopeptide repeat (TPR)-like fold and a short NH(2)-terminal helix. Although it is known that the cytoplasmic domain is necessary for assembly of Mdv1p and Dnm1p into fission complexes, the molecular details of this assembly are not clear. In this study, we provide new evidence that the Fis1p-Mdv1p interaction is direct. Furthermore, we show that conditional mutations in the Fis1p TPR-like domain cause fission complex assembly defects that are suppressed by mutations in the Mdv1p-predicted coiled coil. We also define separable functions for the Fis1p NH(2)-terminal arm and TPR-like fold. These studies suggest that the concave binding surface of the Fis1p TPR-like fold interacts with Mdv1p during mitochondrial fission and that Mdv1p facilitates Dnm1p recruitment into functional fission complexes. PMID- 16247029 TI - Termination of cAMP signals by Ca2+ and G(alpha)i via extracellular Ca2+ sensors: a link to intracellular Ca2+ oscillations. AB - Termination of cyclic adenosine monophosphate (cAMP) signaling via the extracellular Ca(2+)-sensing receptor (CaR) was visualized in single CaR expressing human embryonic kidney (HEK) 293 cells using ratiometric fluorescence resonance energy transfer-dependent cAMP sensors based on protein kinase A and Epac. Stimulation of CaR rapidly reversed or prevented agonist-stimulated elevation of cAMP through a dual mechanism involving pertussis toxin-sensitive Galpha(i) and the CaR-stimulated increase in intracellular [Ca2+]. In parallel measurements with fura-2, CaR activation elicited robust Ca2+ oscillations that increased in frequency in the presence of cAMP, eventually fusing into a sustained plateau. Considering the Ca2+ sensitivity of cAMP accumulation in these cells, lack of oscillations in [cAMP] during the initial phases of CaR stimulation was puzzling. Additional experiments showed that low-frequency, long duration Ca2+ oscillations generated a dynamic staircase pattern in [cAMP], whereas higher frequency spiking had no effect. Our data suggest that the cAMP machinery in HEK cells acts as a low-pass filter disregarding the relatively rapid Ca2+ spiking stimulated by Ca(2+)-mobilizing agonists under physiological conditions. PMID- 16247030 TI - Retinal ganglion cell degeneration is topological but not cell type specific in DBA/2J mice. AB - Using a variety of double and triple labeling techniques, we have reevaluated the death of retinal neurons in a mouse model of hereditary glaucoma. Cell-specific markers and total neuron counts revealed no cell loss in any retinal neurons other than the ganglion cells. Within the limits of our ability to define cell types, no group of ganglion cells was especially vulnerable or resistant to degeneration. Retrograde labeling and neurofilament staining showed that axonal atrophy, dendritic remodeling, and somal shrinkage (at least of the largest cell types) precedes ganglion cell death in this glaucoma model. Regions of cell death or survival radiated from the optic nerve head in fan-shaped sectors. Collectively, the data suggest axon damage at the optic nerve head as an early lesion, and damage to axon bundles would cause this pattern of degeneration. However, the architecture of the mouse eye seems to preclude a commonly postulated source of mechanical damage within the nerve head. PMID- 16247031 TI - Mitogen-inducible gene 6 is an endogenous inhibitor of HGF/Met-induced cell migration and neurite growth. AB - Hepatocyte growth factor (HGF)/Met signaling controls cell migration, growth and differentiation in several embryonic organs and is implicated in human cancer. The physiologic mechanisms that attenuate Met signaling are not well understood. Here we report a mechanism by which mitogen-inducible gene 6 (Mig6; also called Gene 33 and receptor-associated late transducer) negatively regulates HGF/Met induced cell migration. The effect is observed by Mig6 overexpression and is reversed by Mig6 small interfering RNA knock-down experiments; this indicates that endogenous Mig6 is part of a mechanism that inhibits Met signaling. Mig6 functions in cells of hepatic origin and in neurons, which suggests a role for Mig6 in different cell lineages. Mechanistically, Mig6 requires an intact Cdc42/Rac interactive binding site to exert its inhibitory action, which suggests that Mig6 acts, at least in part, distally from Met, possibly by inhibiting Rho like GTPases. Because Mig6 also is induced by HGF stimulation, our results suggest that Mig6 is part of a negative feedback loop that attenuates Met functions in different contexts and cell types. PMID- 16247032 TI - Merlin/NF-2 mediates contact inhibition of growth by suppressing recruitment of Rac to the plasma membrane. AB - Introduction of activated p21-activated kinase (PAK) is sufficient to release primary endothelial cells from contact inhibition of growth. Confluent cells display deficient activation of PAK and translocation of Rac to the plasma membrane at matrix adhesions. Targeting Rac to the plasma membrane rescues these cells from contact inhibition. PAK's ability to release human umbilical vein endothelial cells from contact inhibition is blocked by an unphosphorylatable form of its target Merlin, suggesting that PAK promotes mitogenesis by phosphorylating, and thus inactivating, Merlin. Merlin mutants, which are presumed to exert a dominant-negative effect, enable recruitment of Rac to matrix adhesions and promote mitogenesis in confluent cells. Small interference RNA mediated knockdown of Merlin exerts the same effects. Dominant-negative Rac blocks PAK-mediated release from contact inhibition, implying that PAK functions upstream of Rac in this signaling pathway. These results provide a framework for understanding the tumor suppressor function of Merlin and indicate that Merlin mediates contact inhibition of growth by suppressing recruitment of Rac to matrix adhesions. PMID- 16247033 TI - Cell surface counter receptors are essential components of the unconventional export machinery of galectin-1. AB - Galectin-1 is a component of the extracellular matrix as well as a ligand of cell surface counter receptors such as beta-galactoside-containing glycolipids, however, the molecular mechanism of galectin-1 secretion has remained elusive. Based on a nonbiased screen for galectin-1 export mutants we have identified 26 single amino acid changes that cause a defect of both export and binding to counter receptors. When wild-type galectin-1 was analyzed in CHO clone 13 cells, a mutant cell line incapable of expressing functional galectin-1 counter receptors, secretion was blocked. Intriguingly, we also find that a distant relative of galectin-1, the fungal lectin CGL-2, is a substrate for nonclassical export from Chinese hamster ovary (CHO) cells. Alike mammalian galectin-1, a CGL 2 mutant defective in beta-galactoside binding, does not get exported from CHO cells. We conclude that the beta-galactoside binding site represents the primary targeting motif of galectins defining a galectin export machinery that makes use of beta-galactoside-containing surface molecules as export receptors for intracellular galectin-1. PMID- 16247035 TI - Patient page. Fruit and the brain: the more the better! PMID- 16247034 TI - The mechanisms and dynamics of (alpha)v(beta)3 integrin clustering in living cells. AB - During cell migration, the physical link between the extracellular substrate and the actin cytoskeleton mediated by receptors of the integrin family is constantly modified. We analyzed the mechanisms that regulate the clustering and incorporation of activated alphavbeta3 integrins into focal adhesions. Manganese (Mn2+) or mutational activation of integrins induced the formation of de novo F actin-independent integrin clusters. These clusters recruited talin, but not other focal adhesion adapters, and overexpression of the integrin-binding head domain of talin increased clustering. Integrin clustering required immobilized ligand and was prevented by the sequestration of phosphoinositole-4,5 bisphosphate (PI(4,5)P2). Fluorescence recovery after photobleaching analysis of Mn(2+)-induced integrin clusters revealed increased integrin turnover compared with mature focal contacts, whereas stabilization of the open conformation of the integrin ectodomain by mutagenesis reduced integrin turnover in focal contacts. Thus, integrin clustering requires the formation of the ternary complex consisting of activated integrins, immobilized ligands, talin, and PI(4,5)P2. The dynamic remodeling of this ternary complex controls cell motility. PMID- 16247036 TI - Plantar reflex amusement: misuse, ruse, disuse, and abuse. PMID- 16247037 TI - Narcolepsy: selective hypocretin (orexin) neuronal loss and multiple signaling deficiencies. PMID- 16247038 TI - Improving patient safety: what can detailed case analysis tell us? PMID- 16247039 TI - Rotational vertebral artery occlusion: a clinical entity or various syndromes? PMID- 16247040 TI - Should the Babinski sign be part of the routine neurologic examination? AB - BACKGROUND: The Babinski sign is a well-known sign of upper motor neuron dysfunction that is widely considered an essential element of a complete neurologic examination. Little is known about reliability and validity of this sign. A less well-known sign of upper motor neuron dysfunction, decreased speed of foot tapping, also has not been carefully evaluated. Scientific evaluation of findings of the physical examination is crucial in directing busy clinicians. METHODS: Ten physicians (five neurologists and five non-specialists) examined each foot of 10 subjects, 8 of whom had known unilateral upper motor neuron weakness, 1 had bilateral leg weakness secondary to ALS, and 1 had no known neurologic deficits. Our main outcome measures were inter-rater reliability (kappa values) and accuracy (agreement with known upper motor neuron weakness). RESULTS: The reliability of the Babinski sign was fair (kappa 0.30) and was substantial for foot tapping (kappa 0.73). Agreement with known weakness was 56% for Babinski sign and 85% for foot tapping. Reliability and accuracy for both tests were similar for neurologists and non-specialists. CONCLUSIONS: The interobserver reliability and validity of the Babinski sign for identifying upper motor neuron weakness are limited. Slowness of foot tapping may be a more useful sign. PMID- 16247041 TI - Predictors of major neurologic improvement after thrombolysis in acute stroke. AB - BACKGROUND: Major neurologic improvement at 24 hours after administration of recombinant tissue plasminogen activator (rt-PA) in acute stroke may predict good outcome at 3 months. OBJECTIVE: To identify predictors of major neurologic improvement at 24 hours after IV rt-PA administration and its relationship with outcome at 3 months. METHODS: The authors analyzed patients with acute stroke treated with IV rt-PA from two academic centers in London, Ontario, and 33 affiliated hospitals between 1999 and 2003. Major neurologic improvement was defined by a > or = 8-point improvement in NIH Stroke Scale (NIHSS) score or an NIHSS score of 0 or 1 at 24 hours. Good outcome was defined as a 3-month modified Rankin Scale of 0 to 1. RESULTS: Of 219 patients with acute stroke treated with rt-PA, 61 (28%) had major neurologic improvement at 24 hours. Glucose levels < 8 mmol/L (OR 4.98, 95% CI 1.6 to 15.2), lack of cortical involvement on 24 hour CT scan (OR 3.97, 95% CI 1.87 to 8.43), and female sex (OR 2.4, 95% CI 1.12 to 5.13) were associated with major neurologic improvement after adjusting for covariates. Patients with major neurologic improvement had a shorter hospital stay (6.7 vs 14.3 days; p = 0.001). Major neurologic improvement was an independent predictor of good outcome at 3 months (OR 12.8, 95% CI 4.72 to 34.6). CONCLUSIONS: Major neurologic improvement after rt-PA was observed in 28% of patients and independently predicted good outcome at 3 months. Female sex, glucose levels < 8 mmol/L, and absence of cortical involvement at 24 hours CT scan were associated with major neurologic improvement. PMID- 16247042 TI - Clinical importance of microbleeds in patients receiving IV thrombolysis. AB - BACKGROUND: Cerebral microbleeds (MBs) detected on gradient echo (GRE) imaging may be a risk factor for hemorrhagic complications in patients with stroke treated with IV tissue plasminogen activator (tPA). METHODS: The authors prospectively evaluated patients with acute ischemic stroke treated with IV tPA between 3 and 6 hours of symptom onset. MRI scans, including GRE imaging, were performed prior to tPA treatment, 3 to 6 hours after treatment and at day 30. The authors compared the frequency of hemorrhagic complications after thrombolysis in patients with and without MBs on their baseline GRE imaging. RESULTS: Seventy consecutive patients (mean age, 71 +/- 29 years; 31 men, 39 women) were included. MBs were identified in 11 patients (15.7%) on baseline GRE imaging. There was no significant difference in the frequency of either symptomatic or asymptomatic hemorrhagic complications after thrombolysis between patients with and without MBs at baseline. None of the 11 patients with MBs (0%) at baseline had a symptomatic intracerebral hemorrhage compared with 7 of 59 patients who did not have baseline MBs (11.9%). In addition, no patients with baseline MBs had asymptomatic hemorrhagic transformation observed at the site of any pre-treatment MB. CONCLUSIONS: The presence of cerebral microbleeds on gradient echo imaging does not appear to substantially increase the risk of either symptomatic or asymptomatic brain hemorrhage following IV tissue plasminogen activator administered between 3 and 6 hours after stroke onset. PMID- 16247043 TI - Initial emergency department blood pressure as predictor of survival after acute ischemic stroke. AB - OBJECTIVE: To evaluate the association of diastolic blood pressure (dBP), systolic blood pressure (sBP), and mean arterial pressure (MAP) in the emergency department (ED) with mortality within 90 days in patients with acute ischemic stroke (AIS). METHODS: This observational study was conducted at an ED with an annual census of 70,000 visits, with approximately 500 for AIS. The cohort consisted of 357 patients who presented to the ED within 24 hours of stroke symptom onset. sBP and dBP were measured at triage by a nurse blinded to the study. The duration of follow-up was limited to the first 90 days following ED presentation. BP levels were categorized as low, normotensive, and high after examining scatter plots of a patient's risk of death adjusted for duration of follow-up vs dBP, sBP, and MAP. These BP categories were evaluated using indicator variables in Cox proportional hazards models, after adjusting for age, sex, and the National Institute of Health Stroke Scale (NIHSS) score. RESULTS: Patients with low BP (dBP <70, sBP <155, or MAP <100 mm Hg) were significantly more likely to die within 90 days than those with BP in the normotensive range (dBP 70 to 105, sBP 155 to 220, MAP 100 to 140 mm Hg). These associations were significant even after adjusting for age, gender, and NIHSS score. CONCLUSIONS: There appears to be an optimal acute BP range below which early mortality is greater following AIS, suggesting avoidance of hypotension in the first 24 hours. PMID- 16247045 TI - Fruit and vegetable consumption and risk of stroke: a meta-analysis of cohort studies. AB - BACKGROUND: Fruit and vegetable consumption is associated with lower rates of coronary heart disease. Results from observational studies suggest a similar association with stroke. OBJECTIVE: To assess the evidence from prospective observational studies on fruit and vegetable intake and risk of stroke. METHODS: A meta-analysis of prospective studies was conducted to examine the association between fruit and vegetable intake and stroke. Studies were selected if they reported relative risk (RR) and 95% CI for any type of stroke and used a validated questionnaire for food intake assessment. Pooled RR were calculated and linearity of the associations was examined. RESULTS: Seven studies were eligible for the meta-analysis, including 90,513 men, 141,536 women, and 2,955 strokes. The risk of stroke was decreased by 11% (RR 95% CI: 0.89 [0.85 to 0.93]) for each additional portion per day of fruit, by 5% (RR: 0.95 [0.92 to 0.97]) for fruit and vegetables, and by 3% (RR: 0.97 [0.92 to 1.02]; NS) for vegetables. The association between fruit or fruit and vegetables and stroke was linear, suggesting a dose-response relationship. CONCLUSIONS: This meta-analysis of cohort studies suggests that fruit and fruit and vegetable consumption decreases the risk of stroke. PMID- 16247044 TI - Concomitant loss of dynorphin, NARP, and orexin in narcolepsy. AB - BACKGROUND: Narcolepsy with cataplexy is associated with a loss of orexin/hypocretin. It is speculated that an autoimmune process kills the orexin producing neurons, but these cells may survive yet fail to produce orexin. OBJECTIVE: To examine whether other markers of the orexin neurons are lost in narcolepsy with cataplexy. METHODS: We used immunohistochemistry and in situ hybridization to examine the expression of orexin, neuronal activity-regulated pentraxin (NARP), and prodynorphin in hypothalami from five control and two narcoleptic individuals. RESULTS: In the control hypothalami, at least 80% of the orexin-producing neurons also contained prodynorphin mRNA and NARP. In the patients with narcolepsy, the number of cells producing these markers was reduced to about 5 to 10% of normal. CONCLUSIONS: Narcolepsy with cataplexy is likely caused by a loss of the orexin-producing neurons. In addition, loss of dynorphin and neuronal activity-regulated pentraxin may contribute to the symptoms of narcolepsy. PMID- 16247046 TI - Serum lipid levels and in-hospital mortality in patients with intracerebral hemorrhage. AB - OBJECTIVE: To test the hypothesis that low serum cholesterol and low serum triglyceride levels at admission are related to an increase of in-hospital mortality in patients with first-ever supratentorial spontaneous intracerebral hemorrhage (ICH). METHODS: The authors obtained the serum cholesterol and triglyceride levels during the first 48 hours after first-ever ICH in 184 patients. They analyzed the impact of serum cholesterol and triglyceride concentrations on the in-hospital mortality after adjustment for possible confounding variables according to the results of the univariate analysis (age, hemorrhage volume, intraventricular extension, glycemia, serum albumin, and Glasgow Coma Scale score at admission) using the Cox proportional hazards model. They also analyzed the survival curves according to the cholesterol and triglyceride quartiles. RESULTS: Low serum cholesterol (p = 0.002; hazard ratio [HR] 0.988 [95% CI 0.979 to 0.997] mg/dL) and low serum triglyceride (p = 0.011; HR 0.986 [95% CI 0.976 to 0.997] mg/dL) concentrations were independently associated with increased in-hospital mortality after ICH. Analyzed by quartiles, the HR of in-hospital mortality was 3.136 (95% CI 0.833 to 11.087) for patients in the lowest cholesterol quartile (< 166 mg/dL) and 3.484 (95% CI 1.088 to 11.155) for patients in the lowest triglyceride quartile (< 74 mg/dL). CONCLUSIONS: Low serum cholesterol and triglyceride levels obtained during the first hours after intracerebral hemorrhage (ICH) are strong independent predictors of in-hospital mortality in patients with spontaneous supratentorial ICH. PMID- 16247047 TI - Alcohol consumption and cognitive function in late life: a longitudinal community study. AB - OBJECTIVE: To examine the association between alcohol use and cognitive decline in a longitudinal study of a representative elderly community sample free of dementia at baseline. METHODS: Cognitive functions and self-reported drinking habits were assessed at 2-year intervals over an average of 7 years of follow-up. Cognitive measures, grouped into composites, were examined in association with alcohol consumption. Trajectory analyses identified latent homogeneous groups with respect to alcohol use frequency over time, and their association with average decline over the same period in each cognitive domain. Models controlled for age, sex, education, depression, smoking, general mental status (Mini-Mental State Examination [MMSE]), performance on the given test at baseline, and subsequent new-onset dementia during follow-up. RESULTS: The authors found three homogeneous trajectories that they characterized as no drinking, minimal drinking, and moderate drinking. Few heavy drinkers were identified in this elderly cohort. Compared to no drinking, both minimal and moderate drinking were associated with lesser decline on the MMSE and Trailmaking tests. Minimal drinking was also associated with lesser decline on tests of learning and naming. These associations were more pronounced when comparing current drinkers to former drinkers (quitters) than to lifelong abstainers. CONCLUSION: In a representative elderly cohort over an average of 7 years, a pattern of mild-to-moderate drinking, compared to not drinking, was associated with lesser average decline in cognitive domains over the same period. PMID- 16247048 TI - Decline in cognitive and functional skills increases mortality risk in nondemented elderly. AB - OBJECTIVE: To investigate the relation between rate of decline in cognitive and functional/physical abilities and risk of death in nondemented elderly. METHODS: Data were included from individuals participating in a prospective study of aging and dementia in Medicare recipients, 65 years and older, residing in northern Manhattan. The authors included 878 members of the cohort who had measures of memory, cognitive, language, or functional scores over three study intervals, excluding all participants who were demented or had more than one problem in activity of daily living (ADL) skills at baseline. Participants were classified as showing no decline, slow, medium, or rapid rate of decline, based on the slope of change in cognitive and functional/physical factors. The authors used survival methods to examine the relation of rate of decline in cognitive and functional performance to subsequent mortality in younger and older nondemented elderly and across three ethnic groups, adjusting for potential confounders. RESULTS: Nondemented elderly with preserved ADL skills who showed rapid rates of decline on measures of visuospatial reasoning/cognitive, language, ADL, and instrumental ADL functions were approximately twice as likely to die as nondemented elderly who showed no decline or slower rates of decline, while rate of decline in memory or in measures of extremity mobility was not related to risk of death. The association of the rate of decline to risk of death was stronger in relatively young (< or =75 years) than in older participants. CONCLUSIONS: Rate of decline in cognitive and functional skills predicts mortality in nondemented elderly. PMID- 16247049 TI - Brain atrophy rates predict subsequent clinical conversion in normal elderly and amnestic MCI. AB - OBJECTIVE: To test the hypothesis that the atrophy rate measured from serial MRI studies is associated with time to subsequent clinical conversion to a more impaired state in both cognitively healthy elderly subjects and in subjects with amnestic mild cognitive impairment (MCI). METHODS: Ninety-one healthy elderly patients and 72 patients with amnestic MCI who met inclusion criteria were identified from the Mayo Alzheimer's Disease Research Center and Alzheimer's Disease Patient Registry. Atrophy rates of four different brain structures- hippocampus, entorhinal cortex, whole brain, and ventricle--were measured from a pair of MRI studies separated by 1 to 2 years. The time of the second scan marked the beginning of the clinical observation period. RESULTS: During follow-up, 13 healthy patients converted to MCI or Alzheimer disease (AD), whereas 39 MCI subjects converted to AD. Among those healthy at baseline, only larger ventricular annual percent volume change (APC) was associated with a higher risk of conversion (hazard ratio for a 1-SD increase 1.9, p = 0.03). Among MCI subjects, both greater ventricular volume APC (hazard ratio for a 1-SD increase 1.7, p < 0.001) and greater whole brain APC (hazard ratio for a 1-SD increase 1.4, p = 0.007) increased the risk of conversion to AD. Both ventricular APC (hazard ratio for a 1-SD increase 1.59, p = 0.001) and whole brain APC (hazard ratio for a 1-SD increase 1.32, p = 0.009) provided additional predictive information to covariate-adjusted cross-sectional hippocampal volume at baseline about the risk of converting from MCI to AD. DISCUSSION: Higher whole brain and ventricle atrophy rates 1 to 2 years before baseline are associated with an increased hazard of conversion to a more impaired state. Combining a measure of hippocampal volume at baseline with a measure of either whole brain or ventricle atrophy rates from serial MRI scans provides complimentary predictive information about the hazard of subsequent conversion from mild cognitive impairment to Alzheimer disease. However, overlap among those who did vs those who did not convert indicate that these measures are unlikely to provide absolute prognostic information for individual patients. PMID- 16247050 TI - Verbal and visuospatial deficits in dementia with Lewy bodies. AB - OBJECTIVE: To investigate the cognitive decline in dementia with Lewy bodies (DLBs) and characterize the contribution of Lewy bodies (LBs) to cognitive impairment in the presence of concurrent Alzheimer disease (AD). METHODS: Cognitive deficits and rates of progression attributable to DLB and AD neuropathology were investigated in three groups of participants from the longitudinal cohort of the Alzheimer Disease Research Center at Washington University with autopsy-confirmed diagnoses of pure DLB (n = 9), mixed DLB/AD (n = 57), and pure AD (n = 66). Factor analysis was used to recover latent constructs in a comprehensive psychometric test battery, analysis of variance was used to test group differences on the observed dimensions, and random effects models were used to test longitudinal rates of cognitive decline. RESULTS: Patients with AD pathology performed worse on the verbal memory dimension. Patients with LB pathology performed worse on the visuospatial dimension. Combined pathology affected visuospatial performance but not verbal memory. The rate of cognitive decline in the DLB, DLB/AD combined, and the pure AD groups was equivalent. CONCLUSIONS: The comorbid presence of DLB and AD alters the cognitive presentation of visuospatial deficits in dementia but does not alter dementia progression. Both visuospatial and verbal abilities declined at similar rates across the three patient groups. DLB diagnosis may be improved, particularly when there is comorbid AD, by using domain-specific testing. PMID- 16247051 TI - Cognitive profile of patients with newly diagnosed Parkinson disease. AB - OBJECTIVE: To determine the frequency and pattern of cognitive dysfunction in patients with newly diagnosed Parkinson disease (PD) and to identify its demographic and clinical correlates. METHODS: A cohort of 115 consecutive patients with newly diagnosed PD and 70 healthy controls underwent a comprehensive neuropsychological assessment including tests of psychomotor speed, attention, language, memory, executive and visuospatial functions, as well as measures of affective status. Patients also received quantitative ratings of motor symptom severity and functional status. Neuropsychological performance of PD patients was compared with that of healthy controls and with available normative data. Independent demographic and clinical predictors of cognitive impairment were identified with multiple logistic regression analysis. RESULTS: Relative to controls, PD patients performed significantly worse on most cognitive measures. However, further analysis revealed that group differences in cognitive performance could mainly be explained by measures of immediate memory and executive function. Comparison with normative data showed that impairments were most frequent on measures of executive function, memory and psychomotor speed. In all, 24% of PD patients (4% of controls) displayed defective performance on at least three neuropsychological tests and were classified as cognitively impaired. Late onset of disease was an independent predictor of cognitive dysfunction in PD. CONCLUSION: Cognitive impairments are common even in newly diagnosed Parkinson disease patients, with deficits being most prominent in the domains of memory and executive functions. Older age at disease onset is likely to be an important determinant of cognitive dysfunction in Parkinson disease. PMID- 16247052 TI - Abnormal speech sound representation in persistent developmental stuttering. AB - OBJECTIVES: To determine whether adults with persistent developmental stuttering (PDS) have auditory perceptual deficits. METHODS: The authors compared the mismatch negativity (MMN) event-related brain potential elicited to simple tone (frequency and duration) and phonetic contrasts in a sample of PDS subjects with that recorded in a sample of paired fluent control subjects. RESULTS: Subjects with developmental stuttering had normal MMN to simple tone contrasts but a significant supratemporal left-lateralized enhancement of this electrophysiologic response to phonetic contrasts. In addition, the enhanced MMN correlated positively with speech disfluency as self-rated by the subjects. CONCLUSIONS: Individuals with persistent developmental stuttering have abnormal permanent traces for speech sounds, and their abnormal speech sound representation may underlie their speech disorder. The link between abnormal speech neural traces of the auditory cortex and speech disfluency supports the relevance of speech perception mechanisms to speech production. PMID- 16247054 TI - Possible increasing risk of multiple sclerosis in Catania, Sicily. AB - OBJECTIVE: To update prevalence and incidence rates of multiple sclerosis (MS) in Catania, Italy during 1990 to 1999 and evaluate their temporal profiles to assess a possible increase in the MS risk in our study population. METHODS: We studied the frequency of MS in Catania, Italy (population of 313,110 as reported in the 2001 census). The primary sources for the case ascertainment were the neurologic and motor-rehabilitation departments, the MS centers, the Italian MS Association, private neurologists, and family doctors. We considered as prevalent and incident cases all patients who satisfied Poser's criteria for clinically definite MS (CDMS), laboratory-supported definite MS (LSDMS), clinically probable MS (CPMS), and laboratory-supported probable MS (LSPMS). RESULTS: We found 288 subjects with MS who had onset of disease before December 31, 1999 (prevalence day) in a population of 313,110 inhabitants. The prevalence rate was 92.0/100,000 (95% CI 81.8 to 103.2) and was higher in women (102.4/100,000) than in men (80.4/100,000). The age-specific prevalence showed a peak in the group aged 35 to 44 years (208.2/100,000). From 1990 to 1999, 155 patients with MS had the clinical onset of the disease. The mean annual incidence was 4.7/100,000 (95% CI 4.0 to 5.5). Age-specific incidence showed a peak in the group aged 25 to 34 years (11.7/100,000). Mean annual incidence for 5-year intervals increased from 3.9/100,000 during 1990 to 1994 to 5.5/100,000 during 1995 to 1999. CONCLUSIONS: Prevalence and incidence rates of multiple sclerosis have further increased during the last decade. PMID- 16247053 TI - Caudate volumes in childhood predict symptom severity in adults with Tourette syndrome. AB - BACKGROUND: Most children with Tourette syndrome (TS) experience a marked decline in the severity of tic symptoms during adolescence. Currently no clinical measures can predict whose tic symptoms will persist into adulthood. Previous cross-sectional imaging studies have identified reduced caudate nucleus volumes in subjects with TS. OBJECTIVE: To evaluate whether caudate nucleus volumes in childhood can predict the severity of tic or obsessive-compulsive symptoms at follow-up in early adulthood. METHODS: In a prospective longitudinal study, clinical status and basal ganglia volumes of 43 children with TS were measured on high-resolution magnetic resonance images before age 14 years. Follow-up clinical assessments were conducted after age 16 years, an average of 7.5 years later. Linear regression and Tobit regression analyses were used to assess the association of basal ganglia volumes measured in childhood with the severity of tic and obsessive-compulsive disorder (OCD) symptoms at the time of childhood MRI and at follow-up in early adulthood. RESULTS: Volumes of the caudate nucleus correlated significantly and inversely with the severity of tic and OCD symptoms in early adulthood. Caudate volumes did not correlate with the severity of symptoms at the time of the MRI scan. CONCLUSIONS: Caudate volumes in children with Tourette syndrome predict the severity of tic and obsessive-compulsive symptoms in early adulthood. This study provides compelling evidence that morphologic disturbances of the caudate nucleus within cortico-striatal-thalamo cortical circuits are central to the persistence of both tics and obsessive compulsive symptoms into adulthood. PMID- 16247055 TI - Multidisciplinary ALS care improves quality of life in patients with ALS. AB - OBJECTIVE: To examine the effect of multidisciplinary ALS care on the quality-of life (QoL) in patients with ALS and their caregivers. METHODS: In a cross sectional study, 208 patients with ALS and their caregivers were interviewed. QoL was assessed using the 36-item Short Form Health Survey (SF-36) and two visual analogue scales (VAS). Criteria for multidisciplinary ALS care were: an ALS team headed by a consultant in rehabilitation medicine and consisting of at least a physical therapist, occupational therapist, speech pathologist, dietician and a social worker; use of the Dutch ALS consensus guidelines for ALS care; and at least six incident ALS patients per year. RESULTS: Clinical characteristics and functional loss of the 133 patients receiving multidisciplinary ALS care and the 75 patients receiving general ALS care were similar. The percentage of patients with adequate aids and appliances was higher in those with multidisciplinary ALS care (93.1 vs 81.3%, p = 0.008), whereas the number of visits to professional caregivers was similar in both groups. Patients in the multidisciplinary ALS care group had a better mental QoL on the SF-36 Mental Summary Score than those in the general care group (p = 0.01). The difference in QoL was most pronounced in the domains of Social Functioning and Mental Health, and was independent of the presence of aids and appliances. No significant differences were found in the SF 36 Physical Summary Score, VAS, or in QoL of caregivers of patients with ALS. CONCLUSION: High standard of care improves mental quality-of-life in patients with ALS. PMID- 16247056 TI - Abnormal forebrain activity in functional bowel disorder patients with chronic pain. AB - BACKGROUND: Abnormal cortical pain responses in patients with fibromyalgia and conversion disorder raise the possibility of a neurobiologic basis underlying so called "functional" chronic pain. OBJECTIVE: To use percept-related fMRI to test the hypothesis that patients with a painful functional bowel disorder do not process visceral input or sensations normally or effectively at the cortical level. METHODS: Eleven healthy subjects and nine patients with irritable bowel syndrome (IBS) underwent fMRI during rectal distensions that elicited either a moderate level of urge to defecate or pain. Subjects continuously rated their rectal stimulus-evoked urge or pain sensations during fMRI acquisition. fMRI data were interrogated for activity related to stimulus presence and to specific sensations. RESULTS: In IBS, abnormal responses associated with rectal-evoked sensations were identified in five brain regions. In primary sensory cortex, there were urge-related responses in the IBS but not control group. In the medial thalamus and hippocampus, there were pain-related responses in the IBS but not control group. However, pronounced urge- and pain-related activations were present in the right anterior insula and the right anterior cingulate cortex in the control group but not the IBS group. CONCLUSIONS: Percept-related fMRI revealed abnormal urge- and pain-related forebrain activity during rectal distension in patients with irritable bowel syndrome (IBS). As visceral stimulation evokes pain and triggers unconscious processes related to homeostasis and reflexes, abnormal brain responses in IBS may reflect the sensory symptoms of rectal pain and hypersensitivity, visceromotor dysfunction, and abnormal interoceptive processing. PMID- 16247057 TI - Left caloric vestibular stimulation ameliorates right hemianesthesia. AB - BACKGROUND: Left caloric vestibular stimulation (CVS) transiently reduces impairments of right-brain-damaged patients with left unilateral neglect, including left hemianesthesia, contralateral to the side of the lesion (contralesional). Conversely, no effect on right contralesional hemianesthesia in left-brain-damaged patients is seen with right CVS. This discrepancy is unexplained. METHODS: The authors explored the effect of CVS on right- and left brain-damaged patients with hemianesthesia. One left-brain-damaged patient had an fMRI study during tactile stimulation before and after left CVS. The same fMRI touch study, without CVS, was performed in neurologically unimpaired subjects. RESULTS: A transient remission of right hemianesthesia associated with left brain damage was observed, provided that cold CVS was administered to the left ear. In the left-brain-damaged patient studied with fMRI, left CVS modulated the neural response to right hand tactile stimuli of a portion of the secondary somatosensory area (SII) of the right hemisphere. In neurologically unimpaired subjects, fMRI scans showed that the same part of area SII in the right hemisphere was activated by ipsilateral right-sided touches and to a larger extent than area SII in the left hemisphere by left-sided touches. CONCLUSIONS: Left caloric vestibular stimulation is effective on both left and right hemianesthesia because it modulates the hemisphere that has a more complete representation of, or is capable to attend to, the whole somatosensory surface of the body. These results suggest a hardwired hemispheric asymmetry in hand representation, starting from a somatotopically organized brain region such as area SII. PMID- 16247058 TI - Neurologic patient safety: an in-depth study of malpractice claims. AB - This in-depth study of neurologic malpractice claims indicated authentic, preventable patient harm in 24 of 42 cases, enabling comparison with larger but administratively abstracted summary reports. Principal findings included the common occurrence of outpatient events, lapses in communication with patients and other providers, the need for follow-through by the consultant neurologist even when not primarily responsible, the frequency of diagnostic errors, and pitfalls associated with imaging. PMID- 16247059 TI - Rotational vertebral artery syndrome: oculographic analysis of nystagmus. AB - In four patients with rotational vertebral artery syndrome (RVAS), the initial nystagmus was mostly downbeat, with the horizontal and torsional components beating toward the compressed vertebral artery side (n = 3) or directed away (n = 1). Three patients showed spontaneous reversal of the nystagmus and two exhibited no or markedly diminished responses on immediate retrial of head rotation (habituation). The patterns of nystagmus suggest that RVAS may result from differing mechanisms. PMID- 16247060 TI - A synovial cyst in the cervical spine causing acute spinal cord compression. PMID- 16247061 TI - Medial medullary infarction: abnormal ocular motor findings. AB - In 20 consecutive patients with isolated medial medullary infarction, abnormal ocular motor findings included nystagmus (n = 8), ocular contrapulsion (n = 5), and contralesional ocular tilt reaction (n = 2). The nystagmus was ipsilesional (n = 4), gaze-evoked (n = 5), upbeating (n = 4), and hemiseesaw (n = 1). The ocular motor abnormalities may be explained by involvements of the nucleus prepositus hypoglossi, medial longitudinal fasciculus or efferent fibers from the vestibular nuclei, climbing fibers, and cells of the paramedian tracts. PMID- 16247062 TI - Self-treatment for benign paroxysmal positional vertigo of the posterior semicircular canal. AB - The authors compared the efficacy of the Epley procedure with and without self treatment in 80 patients with benign paroxysmal positional vertigo of the posterior semicircular canal. The self-treatment group (88%) had better results than the Epley-alone group (69%) based on both symptoms and nystagmus (p = 0.048). Complications occurred in 2.6% of the Epley-alone group and in 7.5% of the self-treatment group (p = 0.62). PMID- 16247063 TI - Early-onset toe walking in rippling muscle disease due to a new caveolin-3 gene mutation. AB - The authors describe a family with autosomal dominant rippling muscle disease (RMD) and prominent early-onset toe walking. Molecular analysis revealed a novel heterozygous G > A transition at nucleotide position 136 in exon 2 of the caveolin-3 gene (CAV3). The role of Achilles tendon lengthening in more severe forms of RMD is discussed. PMID- 16247064 TI - Inclusion body myopathy and Paget disease is linked to a novel mutation in the VCP gene. AB - Mutations in the valosin-containing protein (VCP) on chromosome 9p13-p12 were recently found to be associated with hereditary inclusion body myopathy, Paget disease of the bone, and frontotemporal dementia (IBMPFD). We identified a novel missense mutation in the VCP gene (R159H; 688G>A) segregating with this disease in an Austrian family of four affected siblings, who exhibited progressive proximal myopathy and Paget disease of the bone but without clinical signs of dementia. PMID- 16247066 TI - Hypokalemic weakness in hyperaldosteronism: activity-dependent conduction block. AB - The authors describe a 48-year-old man who presented with acute weakness. Serum K+ was 1.7 mmol/L, and investigations established hyperaldosteronism. Nerve excitability studies during hypokalemia demonstrated that axons were of high threshold with a fanning out of threshold electrotonus, consistent with hyperpolarization. Activity-dependent conduction block was induced by voluntary contraction. Excitability abnormalities resolved with K+ replacement. Activity dependent conduction block induced by normal activity may contribute to weakness and paralysis developing with hypokalemia. PMID- 16247065 TI - An open study of botulinum-A toxin treatment of trigeminal neuralgia. AB - Thirteen subjects with trigeminal neuralgia were treated with botulinum-A neurotoxin (BoNT/A) in an open-label pilot study. After BoNT/A, visual analog scale score, surface area of pain, and therapeutic coefficient were reduced in all patients and for all branch trigeminal nerves studied. Therefore, BoNT/A is an efficient treatment. There were no major side effects. A placebo-controlled clinical trial is needed to confirm these findings. PMID- 16247067 TI - Posterior circulation infarct after bronchial artery embolization and coiling. PMID- 16247068 TI - Identifying seizure clusters in patients with epilepsy. AB - Clinicians often encounter patients whose neurologic attacks appear to cluster. In a daily diary study, the authors explored whether clustering is a true phenomenon in epilepsy and can be identified in the clinical setting. Nearly half the subjects experienced at least one episode of three or more seizures in 24 hours; 20% also met a statistical clustering criterion. Utilizing the clinical definition of clustering should identify all seizure clusterers, and false positives can be determined with diary data. PMID- 16247069 TI - Treatment delay and the risk of prolonged status epilepticus. AB - Factors contributing to the duration of a single convulsive seizure > 5 minutes were analyzed in 157 children. The medically treated episodes were compared with seizure episodes resolving without treatment (n = 27). Major differences were in age (p = 0.016) and etiology (p = 0.003), and the association between treatment delay and response became significant after 30 minutes when this was analyzed as a single variable (p = 0.003) in Cox regression analysis. PMID- 16247071 TI - Migrating intraventricular cysticercus during MRI. PMID- 16247070 TI - Analysis of LRRK2 functional domains in nondominant Parkinson disease. AB - A comprehensive sequence analysis of 29 exons that code for the functional domains of LRRK2 in 160 nondominant Parkinson disease (PD) patients was performed. Novel variant screening in a further 470 sporadic PD patients and 630 controls revealed two novel variants (R1067Q and IVS33 + 6 T>A), which are likely to be pathogenic in five patients. One patient presented initially with a typical essential tremor phenotype, expanding the phenotypic spectrum of LRRK2 mutations. PMID- 16247072 TI - Large cerebral artery involvement in CADASIL. AB - The authors evaluated the involvement of large cerebral artery in 13 patients with cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) with angiography (12 MR and one conventional). Five patients (38%) showed stenosis: at the middle cerebral artery in three, vertebral artery in one, and the internal carotid artery in one. The stenosis persisted on follow-up angiogram in two patients. There were no differences in risk factors between patients with angiographic abnormality and those without, suggesting occasional involvement of large vessels in CADASIL. PMID- 16247073 TI - Results of intervention for lupus patients with self-perceived cognitive difficulties. AB - The authors developed an 8-week psychoeducational group intervention for patients with systemic lupus erythematosus (SLE) who reported cognitive dysfunction but were not globally impaired on neuropsychological testing. Results of a nonrandomized, uncontrolled pilot study of this program in 17 women with SLE suggest that metamemory and memory self-efficacy improve after participation. One hundred percent retention throughout the study further suggests that patients with SLE are willing and capable of successfully completing the program. PMID- 16247074 TI - Scanning speech and word history. PMID- 16247075 TI - Seronegative Epstein-Barr virus myeloradiculitis in an immunocompetent 72-year old woman. PMID- 16247076 TI - Sensory symptoms in acquired neuromyotonia. PMID- 16247077 TI - Episodic hyperhidrosis as the only clinical manifestation of neuromyotonia. PMID- 16247078 TI - ALS syndrome in HTLV-I infection. PMID- 16247079 TI - Reversible anorgasmia with topiramate therapy for migraine. PMID- 16247080 TI - Migraine and somnambulism. PMID- 16247081 TI - RAS pathway activation and an oncogenic RAS mutation in sporadic pilocytic astrocytoma. PMID- 16247082 TI - Personal history: love, death, and the brainstem. PMID- 16247083 TI - A trial of metoclopramide vs sumatriptan for the emergency department treatment of migraines. PMID- 16247084 TI - Long-term outcome of endovascular stenting for symptomatic basilar artery stenosis. PMID- 16247085 TI - Chronic inflammatory demyelinating polyradiculoneuropathy: MRI study of brain and spinal cord. PMID- 16247090 TI - India's war on polio. PMID- 16247086 TI - NINDS AIREN neuroimaging criteria do not distinguish stroke patients with and without dementia. PMID- 16247091 TI - Dean of medicine becomes U of T president. PMID- 16247092 TI - Advisers to the nation. PMID- 16247094 TI - What is the role of dipyridamole in long-term secondary prevention after an ischemic stroke or transient ischemic attack? PMID- 16247095 TI - Spontaneous aortic thrombosis and embolization: antithrombin deficiency and the work-up of hypercoagulable states. PMID- 16247096 TI - Recurrent left-leg venous thrombosis in a woman despite a therapeutic international normalized ratio. PMID- 16247097 TI - Lymphocytic choriomeningitis virus. PMID- 16247098 TI - Can coronary calcification measured by CT predict future coronary events? PMID- 16247099 TI - Efficacy of an extract of North American ginseng containing poly-furanosyl pyranosyl-saccharides for preventing upper respiratory tract infections: a randomized controlled trial. AB - BACKGROUND: Upper respiratory tract infections are a major source of morbidity throughout the world. Extracts of the root of North American ginseng (Panax quinquefolium) have been found to have the potential to modulate both natural and acquired immune responses. We sought to examine the efficacy of an extract of North American ginseng root in preventing colds. METHODS: We conducted a randomized, double-blind, placebo-controlled study at the onset of the influenza season. A total of 323 subjects 18-65 years of age with a history of at least 2 colds in the previous year were recruited from the general population in Edmonton, Alberta. The participants were instructed to take 2 capsules per day of either the North American ginseng extract or a placebo for a period of 4 months. The primary outcome measure was the number of Jackson-verified colds. Secondary variables measured included symptom severity, total number of days of symptoms and duration of all colds. Cold symptoms were scored by subjects using a 4-point scale. RESULTS: Subjects who did not start treatment were excluded from the analysis (23 in the ginseng group and 21 in the placebo group), leaving 130 in the ginseng group and 149 in the placebo group. The mean number of colds per person was lower in the ginseng group than in the placebo group (0.68 [standard deviation (SD) 0.82] v. 0.93 [SD 0.91], difference 0.25%, 95% confidence interval [CI] 0.04-0.45). The proportion of subjects with 2 or more Jackson-verified colds during the 4-month period (10.0% v. 22.8%, 12.8% difference, 95% CI 4.3-21.3) was significantly lower in the ginseng group than in the placebo group, as were the total symptom score (77.5 [SD 84.6] v. 112.3 [SD 102.5], difference 1.5%, 95% CI 1.2-2.0) and the total number of days cold symptoms were reported (10.8 [SD 9.7] v. 16.5 [SD 13.8] days, difference 1.6%, 95% CI 1.3-2.0) for all colds. INTERPRETATION: Ingestion of a poly-furanosyl-pyranosyl-saccharide-rich extract of the roots of North American ginseng in a moderate dose over 4 months reduced the mean number of colds per person, the proportion of subjects who experienced 2 or more colds, the severity of symptoms and the number of days cold symptoms were reported. PMID- 16247100 TI - How should we respond to the highly toxogenic NAP1/ribotype 027 strain of Clostridium difficile? PMID- 16247101 TI - Studies of "natural" remedies for the common cold: pitfalls and pratfalls. PMID- 16247102 TI - The Declaration of Helsinki: an update on paragraph 30. PMID- 16247105 TI - Pharmaceutical advertising. PMID- 16247106 TI - Other advantages to aerobic exercise. PMID- 16247107 TI - HIV/AIDS and leishmaniasis coinfection in Ethiopia. PMID- 16247103 TI - Early and innovative interventions for severe sepsis and septic shock: taking advantage of a window of opportunity. AB - The pathogenic, diagnostic and therapeutic landscape of sepsis is no longer confined to the intensive care unit: many patients from other portals of entry to care, both outside and within the hospital, progress to severe disease. Approaches that have led to improved outcomes with other diseases (e.g., acute myocardial infarction, stroke and trauma) can now be similarly applied to sepsis. Improved understanding of the pathogenesis of severe sepsis and septic shock has led to the development of new therapies that place importance on early identification and aggressive management. This review emphasizes approaches to the early recognition, diagnosis and therapeutic management of sepsis, giving the clinician the most contemporary and practical approaches with which to treat these patients. PMID- 16247108 TI - Well-water maintenance. PMID- 16247110 TI - Are FASD guidelines practical and sustainable? PMID- 16247112 TI - No resting place: African American women at the crossroads of violence. AB - Seeking safe places after leaving abusive relationships is often an intricate process for African American women. Survivor-victims of gender violence frequently experience ongoing trauma because of race, ethnicity, class, sexual orientation, and other stigmatizing social identities. All too often, women of color must handle leaving the gender violence simultaneously with the ongoing threat of cultural violence. The intersection of gender and cultural violence (e.g., racism, discrimination) complicate African American women's ability to obtain and sustain safe environments. These intersections are critical crossroads in African American women's lives. The results of this womanist and Black feminist study are presented in an alternative way to highlight the primacy of respondents' voices. In addition, suggestions to inform the practice of research with African American women are included. PMID- 16247113 TI - Resilience among urban adolescent mothers living with violence: listening to their stories. AB - Using a risk and resilience approach grounded in a multicultural feminist perspective, this qualitative study explores 10 urban adolescent mothers' experiences with multiple forms of violence, the relationships between violence and school, and their resilience, all within the context of welfare reforms. The findings suggest lives rife with simultaneous, often severe violence, the impacts of witnessing parental violence, linkages between family and partner violence, and cumulative violence exposure as a barrier to positive school outcomes. Several young women emerge as resilient, with five distinct factors conceptualized as contributing to resilience. In conclusion, the implications for welfare policies and programs are discussed. PMID- 16247117 TI - A portrait of interventional radiologists in the United States. AB - OBJECTIVE: In recognition of the emergence of interventional radiology as an important "new component of...radiology," the objective of our study was to provide an extensive and detailed portrait of interventional radiologists, their professional activities, and the practices in which they work. MATERIALS AND METHODS: We tabulated data from the American College of Radiology's 2003 Survey of Radiologists, a stratified random-sample survey that oversampled interventionalists and achieved a 63% response rate with a total of 1,924 responses. Responses were weighted to make them representative of all radiologists in the United States. We compared information about interventionalists with that for other radiologists. RESULTS: Depending on the definition of who is an interventionalist, 8.5-11.5% of radiologists are interventionalists. By most definitions, only slightly under half of interventionalists spend 70% or more of their clinical work time performing interventional procedures. Interventionalists work, on average, 56-58 hr weekly, a few hours longer than other radiologists. The average interventionalist performs procedures in five of the seven categories of procedures into which we divided interventional radiology, compared with one or two categories for other radiologists. The average interventionalist performs procedures in five of the seven broad categories (such as MRI, CT, and nuclear medicine) into which we divided all of radiology, much the same breadth of practice as other subspecialists and also as nonsubspecialists. CONCLUSION: Interventionalists have become a sizable group within radiology. They are in some ways like other radiologists and in other ways different, but they do not spend as much of their time in their subspecialty as some assume and, overall, are not as different. PMID- 16247114 TI - Peritraumatic responses and their relationship to perceptions of threat in female crime victims. AB - Peritraumatic responses, aside from dissociation, have been understudied in acute trauma populations. Participants were 172 female rape, 68 assault, and 80 robbery victims recruited through formal reporting agencies and assessed 1 month after the crime. Despite substantial overlap across crimes, rape victims reported more emotional responses reflecting fear, detachment, shame, and more nonactive behavioral responses. Regression analysis examining the prediction of perceived threat by peritraumatic responses and crime variables indicated that increased duration of crimes; decreased calmness; increased fear; numbing; use of begging, pleading, and crying; and attempts to reason with the perpetrator(s) were all significantly associated with increased appraisal of threat. PMID- 16247118 TI - The predictive value of diagnostic sonography for the effectiveness of conservative treatment of tennis elbow. AB - OBJECTIVE: Tennis elbow is a common complaint. Several treatment strategies have been described, but an optimal strategy has not been identified. Sonographic imaging as a predictive factor has never been studied. The aim of our study was to determine the value of sonographic findings in predicting response to conservative therapy for tennis elbow. This was done in a randomized controlled trial in which the effectiveness of a brace only, physical therapy only, and a combination of both were compared. SUBJECTS AND METHODS: Patients with tennis elbow complaints were randomized. Sonography was performed before randomization in 57 patients. Outcome measures at 6 weeks' follow-up were success rate and decrease in pain (scale, 0-100). Data were analyzed using an intention-to-treat analysis. RESULTS: In only 75% of the imaged patients, sonographic abnormalities were identified and the clinical diagnosis could thus be confirmed. The following entities were identified: hypo- and hyperechogenicity, swelling, calcification, bursitis, enthesopathy, and tendinosis. The positive predictive value of sonography for the different entities varied between 0.78 and 0.82, and the negative predictive value ranged between 0.23 and 0.71. Predictive value was studied by subgroups of sonographic findings: hypoechoic, swelling present, enthesopathy, any entity present, and no entity present. We found no significant differences among the subgroups for either success rate (range, 40-54%) or mean decrease in pain (range, 16-28 percentage points). CONCLUSION: No predictive value of sonography for the detection of abnormalities was identified in this study. Its diagnostic capability showed limited value. However, limitations in this study necessitate drawing definitive conclusions with care. PMID- 16247119 TI - Peroneal nerve palsy associated with knee luxation: evaluation by sonography- initial experiences. AB - OBJECTIVE: Traumatic knee dislocation needs immediate surgical repair to restore joint function. A concomitant traction injury of the peroneal nerve is reported to exist in up to 25% of cases and is often overlooked initially. In patients with major nerve lesions, immediate surgical nerve repair might be necessary to avoid irreversible loss of neural function. In the present study, we tried to evaluate whether sonography is a valuable tool for identification of nerve pathology that warrants surgical intervention. SUBJECTS AND METHODS: In this prospective study, both peroneal nerves in nine patients with one-sided peroneal nerve palsy after closed knee luxation and the peroneal nerves of 11 healthy volunteers were investigated with sonography. Using statistical analysis, we tried to define the comparability and significance of the findings. RESULTS: The mean cross-sectional area of healthy peroneal nerves in the genicular region was 0.18 cm2 (SD, 0.07 cm2). Impaired nerves were significantly discerned because of their increased cross-sectional area at the level of the injury (mean cross sectional area, 0.7 cm2; SD, 0.46 cm2; p < 0.05). Identification of caliber changes and depiction of at least one nerve stump were found to be the most specific criteria for the definition of a major neural lesion. The ability of sonography to provide additional information about surrounding soft-tissue impairment (scar tissue and hematoma formation) proved helpful. CONCLUSION: Sonography allows radiologists to visualize neural and extraneural pathology and to define the exact level and extent of lesions. Thus, it may be a valuable adjunct in the decision of whether surgical intervention is necessary. PMID- 16247120 TI - Visualization of intraarticular structures of the acromioclavicular joint in an ex vivo model using a dedicated MRI protocol. AB - OBJECTIVE: The purpose of this study was to develop an MRI protocol that could visualize the intraarticular structures of the acromioclavicular (AC) joint. MATERIALS AND METHODS: Using six fresh specimens from cadaveric shoulders, several MRI sequences were performed on 1.0-T scanners with a superficial coil (the temporomandibular joint coil). After the radiologic examination, the specimens were prepared for histology and 300-microm-thick, toluidine blue stained sections were prepared that corresponded to the MR images. In each series of sections, immunohistochemistry using a type II collagen antibody was performed to further characterize the intraarticular structures. RESULTS: The coronal 3D T1 weighted fast-field echo water-selective sequence allowed the identification of the intraarticular disk in all cases. Determination on MRI of other intraarticular structures--adipose tissue, synovial fluid, and the borders between neighboring tissues of different types--that corresponded to the histologic sections was possible. The use of a second plane in the 1.0-T sequences did not reveal additional information. CONCLUSION: The described MRI protocol allows the visualization of the intraarticular fibrocartilaginous disk and the border between articular cartilage and the disk. Future clinical studies will indicate the diagnostic value of this protocol. We assume that this MRI protocol could help us to better understand AC joint disorders, in particular those located intraarticularly, and dislocations. PMID- 16247121 TI - Dynamic MRI for distinguishing high-flow from low-flow peripheral vascular malformations. AB - OBJECTIVE: The purpose of our study was to assess the usefulness of dynamic MRI in distinguishing high-flow vascular malformations from low-flow vascular malformations, which do not need angiography for treatment. SUBJECTS AND METHODS: Between September 2001 and January 2003, 16 patients who underwent conventional and dynamic MRI had peripheral vascular malformations (six high- and 10 low flow). The temporal resolution of dynamic MRI was 5 sec. Time intervals between beginning of enhancement of an arterial branch in the vicinity of a lesion in the same slice and the onset of enhancement in the lesion were calculated. We defined these time intervals as "artery-lesion enhancement time." Time intervals between the onset of enhancement in the lesion and the time of the maximal percentage of enhancement above baseline of the lesion within 120 sec were measured. We defined these time intervals as "contrast rise time" of the lesion. Diagnosis of the peripheral vascular malformations was based on angiographic or venographic findings. RESULTS: The mean artery-lesion enhancement time of the high-flow vascular malformations (3.3 sec [range, 0-5 sec]) was significantly shorter than that of the low-flow vascular malformations (8.8 sec [range, 0-20 sec]) (Mann Whitney test, p < 0.05). The mean maximal lesion enhancement time of the high flow vascular malformations (5.8 sec [range, 5-10 sec]) was significantly shorter than that of the low-flow vascular malformations (88.4 sec [range, 50-100 sec]) (Mann-Whitney test, p < 0.01). CONCLUSION: Dynamic MRI is useful for distinguishing high-flow from low-flow vascular malformations, especially when the contrast rise time of the lesion is measured. PMID- 16247122 TI - Is vascularity more evident after exercise? Implications for tendon imaging. AB - OBJECTIVE: The objective of our study was to investigate the effect of activity on tendon vascularity in 17 abnormal patellar tendons. CONCLUSION: Tendon vascularity is significantly increased by activity (p < 0.001). From this finding, we infer that imaging abnormal tendons with color Doppler sonography to detect neovascularization may be most useful after the patient exercises. Investigations to determine how much activity is necessary to ensure maximal vascularity is detected by Doppler sonography are required. PMID- 16247123 TI - Painful stress fractures of the fabella in patients with total knee arthroplasty. AB - OBJECTIVE: The purpose of this study was to document a distinct pattern of stress fractures in the fabella of patients with total knee replacement and to call attention to this unusual complication that causes knee pain in postoperative patients. CONCLUSION: Stress fractures involving the fabella can be a clinically important entity in the patient with persistent or new knee pain after total knee arthroplasty. Scrutiny of radiographs, when coupled with clinical information, allows correct diagnosis of this potentially debilitating injury in symptomatic postoperative patients. PMID- 16247124 TI - Latissimus dorsi tendinosis and tear: imaging features of a pseudotumor of the upper limb in five patients. AB - OBJECTIVE: The objective of our study was to determine the imaging appearances of a pseudotumor of the upper limb, latissimus dorsi tendinosis and tear, in five patients and to correlate those imaging findings with clinical history and histopathology. CONCLUSION: Tears or reactive tendinosis of the latissimus dorsi tendon at its insertion on the proximal humerus may present as a pseudotumor. Awareness of the imaging findings may allow accurate diagnosis and conservative management. PMID- 16247125 TI - Diagnosis of gastric cancer with MDCT using the water-filling method and multiplanar reconstruction: CT-histologic correlation. AB - OBJECTIVE: The purpose of this study was to assess the utility of MDCT with a thin-sliced multiplanar reconstruction (MPR) technique and water-filling method for the diagnosis of gastric cancers. MATERIALS AND METHODS: Sixty-five patients with gastric cancers were preoperatively examined with MDCT using the water filling method. The abdomen was dynamically scanned at 30 and 80 sec after the start of contrast medium administration. MPR images were reconstructed with a slice width of 1.25 mm and a slice interval of 1 mm. The detection rate and accuracy of T staging for gastric cancer were evaluated on MPR images and compared with 5-mm-slice axial images. In addition, MDCT images were correlated with pathologic findings. RESULTS: The detection rate of all gastric cancers using the MPR technique was 65%. The detection rate of advanced gastric cancers was 96.2% (25 of 26), whereas that of early gastric cancers was 41.2% (14 of 34). There was a statistically significant difference (p < 0.05) in the detection rate of early gastric cancers between MPR and 5-mm-slice axial images. The MDCT appearances of gastric cancers were well correlated with pathologic findings such as mucinous component or differences in infiltration of cancer cells. The overall accuracy of CT staging was 85%. MPR images were superior to axial images for the evaluation of the z-axis extent of tumor. CONCLUSION: MDCT with the water-filling method has advantages in acceptable evaluation of depth invasion of gastric carcinomas and in visualization of histologic changes in the tumors. MPR images may be a useful guide for the evaluation of the z-axis extent of tumor. PMID- 16247126 TI - Role of helical CT in diagnosis of gallstone ileus and related conditions. AB - OBJECTIVE: Small-bowel obstruction from gallstone impaction is a pathological entity frequently observed in elderly patients with a history of cholelithiasis or cholecystitis. Diagnostic imaging plays a great role in the management of patients with suspected gallstone ileus and overall in the correct predictive diagnosis: in the last years, some experiences in radiologic diagnosis of this entity by sonography, abdominal plain film and CT, and occasionally MRI have been reported. Some questions related to gallstone ileus are to be considered: one of them is the possibility of recurrence, which increases the operatory risk in these patients. Recurrence may be due either to the presence of overlooked stones that were already in the bowel at the time of surgery but not identified and not removed or to the migration of other stones in patients not previously cholcystectomized. In cases of acute abdomen, establishing an effective conservative therapy may be a critical point. The aim of this retrospective study was to evaluate the capabilities of helical single-detector and MDCT scanners to allow a correct diagnosis of this disease. CONCLUSION: Helical single-detector and MDCT may improve the diagnosis of gallstone ileus, providing important information regarding the exact number, size, and location of ectopic stones and the site of intestinal obstruction or direct visualization of a biliary-enteric fistula, to help clinicians in the therapeutic management of patients. PMID- 16247127 TI - Prospective assessment of interobserver agreement for defecography in fecal incontinence. AB - OBJECTIVE: The primary aim of our study was to determine the interobserver agreement of defecography in diagnosing enterocele, anterior rectocele, intussusception, and anismus in fecal-incontinent patients. The subsidiary aim was to evaluate the influence of level of experience on interpreting defecography. SUBJECTS AND METHODS: Defecography was performed in 105 consecutive fecal-incontinent patients. Observers were classified by level of experience and their findings were compared with the findings of an expert radiologist. The quality of the expert radiologist's findings was evaluated by an intraobserver agreement procedure. RESULTS: Intraobserver agreement was good to very good except for anismus: incomplete evacuation after 30 sec (kappa, 0.55) and puborectalis impression (kappa, 0.54). Interobserver agreement for enterocele and rectocele was good (kappa, 0.66 for both) and for intussusception, fair (kappa, 0.29). Interobserver agreement for anismus: incomplete evacuation after 30 sec was moderate (kappa, 0.47), and for anismus: puborectalis impression was fair (kappa, 0.24). Agreement in grading of enterocele and rectocele was good (kappa, 0.64 and 0.72, respectively) and for intussusception, fair (kappa, 0.39). Agreement separated by experience level was very good for rectocele (kappa, 0.83) and grading of rectoceles (kappa, 0.83) and moderate for intussusception (kappa, 0.44) at the most experienced level. For enterocele and grading, experience level did not influence the reproducibility. CONCLUSION: Reproducibility for enterocele, anterior rectocele, and severity grading is good, but for intussusception is fair to moderate. For anismus, the diagnosis of incomplete evacuation after 30 sec is more reproducible than puborectalis impression. The level of experience seems to play a role in diagnosing anterior rectocele and its grading and in diagnosing intussusception. PMID- 16247129 TI - Effect of adjusted positioning on gastric distention and fluid distribution during CT gastrography. AB - OBJECTIVE: We hypothesized that performing CT gastrography with the patient in the left posterior oblique (LPO) position would provide better distention and less residual fluid for the lower part of the stomach than CT gastrography with the patient in the standard supine position. CONCLUSION: The LPO position for CT guaranteed the distensibility of and minimal residual fluid in the lower part of the stomach, thus resulting in excellent 3D CT gastrographic images. PMID- 16247128 TI - Peroral CT enterography with lactulose solution: preliminary observations. AB - OBJECTIVE: The objective of our study was to evaluate lactulose solution as a new oral contrast agent with the use of peroral CT enterography to determine the adequacy of luminal distention and conspicuity of the bowel wall. CONCLUSION: Peroral CT enterography performed with lactulose solution is a simple and noninvasive method of evaluating the small bowel by obtaining good distention. It can also be used at routine abdominal examinations as a negative contrast agent instead of iodinated oral contrast medium, especially for CT angiography. PMID- 16247130 TI - Incarcerated hernia through the posterior rectus sheath. PMID- 16247131 TI - Focal amyloidoma of the small bowel mimicking adenocarcinoma on CT. PMID- 16247132 TI - Glomus tumor of the stomach: MRI findings. PMID- 16247133 TI - Comparison of IV contrast-enhanced sonography and histopathology of pancreatic cancer. AB - OBJECTIVE: We compared contrast-enhanced sonography findings with pathologic findings in pancreatic cancer to evaluate the ability of contrast-enhanced sonography to depict the pathologic changes associated with pancreatic cancer. SUBJECTS AND METHODS: Thirty-four patients with pancreatic cancer who underwent surgery were investigated. Sonography was performed with contrast material (Levovist) for all patients before surgery. Pathologic findings were evaluated on the basis of the resected cancer specimens. We compared contrast-enhanced sonography findings with pathologic findings. RESULTS: All tumors that were hyperechoic on contrast-enhanced sonography were papillary adenocarcinoma, and all tumors that were hypoechoic on contrast-enhanced sonography were ductal adenocarcinoma. Among ductal adenocarcinomas, five (71.4%) of seven tumors for which the size of the hypoechoic area was unchanged on contrast-enhanced sonography had clear tumor margins with no infiltration or inflammation in the margin. In contrast, all tumors for which the size of the hypoechoic area was reduced on contrast-enhanced sonography had unclear tumor margins with infiltration of cancerous cells and inflammation. Nine (90%) of 10 tumors that showed partial contrast enhancement or a vascular shadow in a hypoechoic area had large or medium-sized vessels within a tumor at pathology. In contrast, only one (4.8%) of 21 tumors that did not show the vascular shadow in a hypoechoic area had no large or medium-sized vessels in a tumor. CONCLUSION: Contrast-enhanced sonography well reflects the pathologic changes of pancreatic cancer and will provide useful information in a pretreatment evaluation. Further studies with a large number of patients will be required to confirm this finding. PMID- 16247134 TI - MRI features of hidradenitis suppurativa and review of the literature. PMID- 16247135 TI - Abdominal imaging features of HELLP syndrome: a 10-year retrospective review. AB - OBJECTIVE: The purpose of this study was to describe the abdominal imaging features associated with HELLP (hemolysis, elevated liver enzymes, low platelets) syndrome, a complication of pregnancy. CONCLUSION: The abdominal imaging features of HELLP syndrome include intraparenchymal and perihepatic hematomas and hemoperitoneum as noted on sonography, CT, and angiography. PMID- 16247136 TI - Fistulous communication between a hepatic hydatid cyst and the gallbladder: diagnosis with MR cholangiopancreatography. PMID- 16247137 TI - MRI of prostate cancer at 1.5 and 3.0 T: comparison of image quality in tumor detection and staging. AB - OBJECTIVE: This prospective study was performed to compare the image quality, tumor delineation, and depiction of staging criteria on MRI of prostate cancer at 1.5 and 3.0 T. SUBJECTS AND METHODS: Twenty-four patients with prostate cancer underwent MRI at 1.5 T using the combined endorectal-body phased-array coil and at 3.0 T using the torso phased-array coil, among them 22 before undergoing radical prostatectomy. The prostate was imaged with T2-weighted sequences in axial and coronal orientations at both field strengths and, in addition, with an axial T1-weighted sequence at 1.5 T. Preoperative analysis of all MR images taken together was compared with the histologic findings to determine the accuracy of MRI for the local staging of prostate cancer. In a retroanalysis, the image quality, tumor delineation, and conspicuity of staging criteria were determined separately for both field strengths and compared. Statistical analysis was performed using Wilcoxon's and the McNemar tests. RESULTS: In the preoperative analysis, MRI (at both 1.5 and 3.0 T) had an accuracy of 73% for the local staging of prostate cancer. The retroanalysis yielded significantly better results for 1.5-T MRI with the endorectal-body phased-array coil in terms of image quality (p < 0.001) and tumor delineation (p = 0.012) than for 3.0-T MRI with the torso phased-array coil. Analysis of the individual staging criteria for extracapsular disease did not reveal a superiority of either of the two field strengths in the depiction of any of the criteria. CONCLUSION: Intraindividual comparison shows that image quality and delineation of prostate cancer at 1.5 T with the use of an endorectal coil in a pelvic phased-array is superior to the higher field strength of 3.0 T with a torso phased-array coil alone. As long as no endorectal coil is available for 3-T imaging, imaging at 1.5 T using the combined endorectal-body phased-array coil will continue to be the gold standard for prostate imaging. PMID- 16247138 TI - MRI of the female pelvis using vaginal gel. AB - OBJECTIVE: Vaginal distention with aqueous gel optimizes MRI evaluation of the vaginal walls and outer contours of the cervix. The objective of this pictorial essay is to illustrate the use of vaginal gel for MRI of the female pelvis. CONCLUSION: Distention of the vagina with gel is an inexpensive, well-tolerated procedure that may improve MRI evaluation of a variety of gynecologic conditions. PMID- 16247139 TI - Female breast radiation exposure during CT pulmonary angiography. AB - OBJECTIVE: The objective of our study was to estimate the effective radiation dose to the female breast during CT pulmonary angiography compared with other routine diagnostic imaging techniques. MATERIALS AND METHODS: We retrospectively reviewed the demographic data of patients who underwent CT pulmonary angiography between May 2000 and December 2002, the diagnostic yield of those studies, and the estimated effective radiation dose to the breast incurred during CT. The estimated effective radiation dose was calculated using the ImPACT CT (Impact Performance Assessment of CT) dosimetry calculator and the CT dose index (CTDI) and was compared with the average glandular dose for two-view screening mammography. RESULTS: During the study period, 1,325 CT pulmonary angiograms were obtained. Sixty percent (797) of the scans were obtained on female patients. The mean age of scanned females was 52.5 years (range, 15-93 years). Of the studies performed in females, 401 (50.31%) were negative, 151 (18.95%) were nondiagnostic, and 245 (30.74%) were positive for pulmonary thromboembolism. The calculated effective minimum dose to the breast of an average 60-kg woman during CT was 2.0 rad (20 mGy) per breast compared with an average glandular dose of 0.300 rad (3 mGy) for standard two-view screening mammography. CONCLUSION: CT pulmonary angiography delivers a minimum radiation dose of 2.0 rad (20 mGy) to the breasts of an average-sized woman. This greatly exceeds the American College of Radiology recommendation of < or = 0.300 rad (3 mGy) or less for standard two view mammography. The potential latent carcinogenic effects of such radiation exposure at this time remain unknown. We encourage the judicious use of CT pulmonary angiography and lower doses and nonionizing radiation alternatives when appropriate. PMID- 16247140 TI - Epipericardial fat necrosis: radiologic diagnosis and follow-up. PMID- 16247141 TI - Birt-Hogg-Dube syndrome: a rare cause of pulmonary cysts. PMID- 16247142 TI - Observer performance in assessing anemia on thoracic CT. AB - OBJECTIVE: The purpose of this study was to evaluate the ability of expert reviewers to differentiate an anemic from a nonanemic state on the basis of visual assessment of the relative attenuation of blood in the left ventricle on noncontrast thoracic CT images and to compare reviewer performance with quantitative measurement of CT density in Hounsfield units. MATERIALS AND METHODS: One hundred two noncontrast thoracic CT examinations were qualitatively reviewed by three independent reviewers. Hounsfield unit measurements of the blood in the left ventricle were recorded by a fourth individual. Anemia was defined as a hemoglobin level of less than 10 g/dL. Receiver operating characteristic (ROC) analyses of expert reviewers were compared with measured Hounsfield units. RESULTS: Hounsfield unit measurements performed significantly better than subjective reviewer analyses for differentiation of an anemic from a nonanemic state (area under ROC curve = 0.85 vs 0.72, 0.70, and 0.69; 95% confidence interval, 0.78-0.92 vs 0.63-0.81, 0.61-0.79, and 0.60-0.78, respectively; p < 0.05). With use of a CT density threshold of 35 H, the sensitivity for anemia was 76% and specificity was 81%, whereas the sensitivity of three reviewers was 40-72% with a specificity of 60-83%. Interobserver agreement was found to be poor by kappa statistic (0.0906-0.2128). The correlation coefficient for the analysis of Hounsfield unit versus hemoglobin level was 0.72. Separating data by patient sex revealed a correlation coefficient of 0.81 for men versus 0.52 for women, although the actual regression lines were not statistically different (p > 0.05). CONCLUSION: Despite expert reviewer analyses, subjective evaluations of blood attenuation characteristics are prone to inaccuracy and show poor interobserver agreement. Quantitative measurements of CT density in Hounsfield units should be performed to accurately differentiate an anemic from a nonanemic state when serum hemoglobin levels are not readily available. PMID- 16247143 TI - Cardiac perforation caused by acrylic cement: a rare complication of percutaneous vertebroplasty. PMID- 16247144 TI - Angiography and dynamic airway evaluation with MDCT in the diagnosis of double aortic arch associated with tracheomalacia. PMID- 16247145 TI - Imaging of the cardiac venous system: comparison of MDCT and conventional angiography. AB - OBJECTIVE: Diagnostic and therapeutic strategies in electrophysiology and interventional cardiology include the coronary venous system. The purpose of this study was to compare MDCT angiography with conventional coronary sinus angiography in terms of detailed anatomic display of the coronary veins. CONCLUSION: MDCT angiography is a reliable alternative to conventional coronary sinus angiography for detailed anatomic display of the coronary veins. PMID- 16247146 TI - MDCT of left anterior descending coronary artery to main pulmonary artery fistula. PMID- 16247147 TI - Interobserver agreement for the interpretation of contrast-enhanced 3D MR angiography and MDCT angiography in peripheral arterial disease. AB - OBJECTIVE: The objective of our study was to compare interobserver agreement for interpretations of contrast-enhanced 3D MR angiography and MDCT angiography in patients with peripheral arterial disease. SUBJECTS AND METHODS: Of 226 eligible patients, 69 were excluded. The remaining 157 consecutive patients were prospectively randomized to either MR angiography (n = 78) or MDCT angiography (n = 79). Two observers independently evaluated for arterial stenosis or occlusion on MR angiography (2,157 segments) and MDCT angiography (2,419 segments) using a 5-point ordinal scale. Vessel wall calcifications were noted. Interobserver agreement for each technique was evaluated with a weighted kappa (kappa(w)) statistic. RESULTS: Although interobserver agreement for both was excellent, the interobserver agreement for MR angiography (kappa(w) = 0.90; 95% confidence interval [CI], 0.89-0.92) was higher than that for MDCT angiography (kappa(w) = 0.85; 95% CI, 0.83-0.86) for reporting the degree of arterial stenosis or occlusion in all segments. For the different anatomic locations, the interobserver agreement for MR angiography versus MDCT angiography was as follows: aortoiliac (kappa(w) =0.91 vs 0.84, respectively), femoropopliteal (kappa(w) = 0.91 vs 0.87), and crural (kappa(w) = 0.90 vs 0.83) segments. The interobserver agreement of MDCT angiography significantly decreased in the presence of calcifications but was still good for all anatomic locations. The lowest agreement was found for crural segments in the presence of calcifications (kappa(w) = 0.67). With MR angiography, there were 12 times more nondiagnostic segments than with MDCT angiography (81 vs 7, respectively). CONCLUSION: Interpretations of MR angiography and MDCT angiography for peripheral arterial disease have an excellent interobserver agreement. MR angiography has a higher interobserver agreement than MDCT angiography, and the presence of calcified segments significantly decreases interobserver agreement for MDCT angiography. PMID- 16247148 TI - MDCT angiography for evaluation of the complete vascular tree of hemodialysis fistulas. AB - OBJECTIVE: The purpose of our study was to assess the clinical feasibility of MDCT angiography for evaluating hemodialysis arteriovenous fistulas (AVFs). MATERIALS AND METHODS: MDCT angiography of the complete vascular trees of 36 failing AVFs or AVF-related complications (20 native and 16 polytetrafluoroethylene graft AVFs) was reviewed. The numbers and degrees of stenoses at the anastomoses, graft loops, and draining and central veins and the presence of aneurysms or thrombosis were recorded. Wilcoxon's signed rank test was used to compare the findings of MDCT angiography with those of digital subtraction angiography (DSA) (n = 10), surgery (n = 22), or both (n = 4) performed within 2-6 days. Kappa statistics were used to correlate the clinical feasibility of MDCT angiography assessed by two reviewers. RESULTS: Among the 14 AVFs examined with both MDCT angiography and DSA, no significant difference was seen in the detection and grading (p = 0.317 to > 0.999) of stenoses at various segments of the entire vascular tree. Among the 36 AVFs examined, MDCT angiography also showed no significant difference from DSA or surgery in revealing vascular stenoses, aneurysms, and thromboses from the supplying artery to central veins (p = 0.317 to > 0.999). Overall, the sensitivity, specificity, positive and negative predictive values, and accuracy of MDCT angiography in lesion detection were 98.7%, 97.5%, 98.8%, 97.2%, and 98.3%, respectively. High image quality with superb interobserver correlation (kappa = 0.809 to > 0.999) validated the clinical feasibility of MDCT angiography for assessing AVFs. CONCLUSION: MDCT angiography is clinically feasible for evaluating the complete vascular tree of failing AVFs and in showing uncommon complications, including brachial aneurysms and central vein lesions. PMID- 16247149 TI - MRI versus helical CT for endoleak detection after endovascular aneurysm repair. AB - OBJECTIVE: The objective of our study was to investigate the diagnostic accuracy of MRI and helical CT for endoleak detection. SUBJECTS AND METHODS: Fifty-two patients underwent endovascular aneurysm repair with nitinol stent-grafts. Follow up data sets included contrast-enhanced biphasic CT and MRI within 48 hr after the intervention; at 3, 6, and 12 months; and yearly thereafter. The endoleak size was categorized as < or = 3%, > 3% < or = 10%, > 10% < or = 30%, or > 30% of the maximum cross-sectional aneurysm area. A consensus interpretation of CT and MRI was defined as the standard of reference. RESULTS: Of 252 data sets, 141 showed evidence for endoleaks. The incidence of types I, II, and III endoleaks and complex endoleaks was 3.2%, 40.1%, 8.7%, and 4.0%, respectively. The sensitivity for endoleak detection was 92.9%, 44.0%, 34.8%, and 38.3% for MRI, biphasic CT, uniphasic arterial CT, and uniphasic late CT, respectively. The corresponding negative predictive values were 91.7%, 58.4%, 54.7%, and 56.1%, respectively. The overall accuracy of endoleak detection and correct sizing was 95.2%, 58.3%, 55.6%, and 57.1% for MRI, biphasic CT, uniphasic arterial CT, and uniphasic late CT, respectively. CONCLUSION: MRI is significantly superior to biphasic CT for endoleak detection and rating of endoleak size, followed by uniphasic late and uniphasic arterial CT scans. MRI shows a significant number of endoleaks in cases with negative CT findings and may help illuminate the phenomenon of endotension. Endoleak rates reported after endovascular aneurysm repair substantially depend on the imaging techniques used. PMID- 16247151 TI - High-resolution 16-MDCT evaluation of radial artery for potential use as coronary artery bypass graft: a feasibility study. AB - OBJECTIVE: The objective of our study was to assess the feasibility of using 16 MDCT angiography for the preoperative assessment of the radial and ulnar arteries and the palmar arches in patients scheduled for radial artery harvesting for coronary artery bypass graft (CABG) surgery. CONCLUSION: Sixteen-MDCT angiography shows promise for the noninvasive preoperative assessment of the radial artery as a CABG donor site. PMID- 16247150 TI - In vivo evaluation of patency and in-stent stenoses after implantation of nitinol stents in iliac arteries using MR angiography. AB - OBJECTIVE: Our study was a prospective in vivo study to evaluate whether MR angiography is suitable for assessing stent patency and grading in-stent stenoses and to examine whether the accuracy of MR angiography changes with time after stent implantation. SUBJECTS AND METHODS: In a prospective study, 34 iliac stenoses in 27 patients were treated by implantation of 35 nitinol stents. MR angiography was performed immediately after stent placement for 32 stents, and both digital subtraction angiography (DSA) and MR angiography were repeated at the 6-month follow-up for 23 stents. Three blinded observers assessed stent patency and the degree of in-stent stenoses on MR angiography and DSA (the standard of reference) images. The difference between the observers' grading of stenoses on DSA and on MR angiography was determined. Statistical analysis was performed using the Student's t test for paired samples. RESULTS: Stent patency was assessed correctly for all stents and both sets of MR angiography images. Evaluation of DSA 1 images (obtained at end of implantation procedure) revealed that 96.9% of in-stent stenoses were less than 50%. On DSA 2 images (obtained at follow-up), 95.7% of in-stent stenoses were graded as less than 50%. The difference between grading of stenoses on DSA and MR angiography images was 15.0% +/- 16.0% (minimum, 0.0%; maximum, 63.3%) for DSA 1 versus MR angiography 1 (statistically significant, p = 0.037) and 9.8% +/- 13.5% (minimum, 0.0%; maximum, 63.3%) for MR angiography 2 versus DSA 2 (not statistically significant, p = 0.355). CONCLUSION: Patency was correctly assessed for all stents on MR angiography. The quality of MR angiography regarding characterization of in-stent stenoses improved with time after stent placement. However, discrepancies of more than 60% between grading of lumen narrowing on DSA and MR angiography images occurred even at the 6-month follow-up. Thus, MR angiography is not yet a reliable technique for characterization of in-stent stenoses. PMID- 16247152 TI - Percutaneous CT-guided multisampling core needle biopsy of thoracic lesions. AB - OBJECTIVE: The purpose of our study was to evaluate the diagnostic yield and the complication rate of percutaneous CT-guided coaxial 18-gauge (1.25-mm diameter) multisampling (five samples) core needle biopsy (CNB) of suspected thoracic lesions. MATERIALS AND METHODS: The records of 75 consecutive patients (29 women, 46 men; age range, 33-92 years) who underwent percutaneous CT-guided adjustable coaxial 18-gauge multisampling (five samples) CNB of a suspected thoracic lesion (eight mediastinal lesions, two chest wall lesions, two pleural lesions, and 63 intrapulmonary lesions) were reviewed. RESULTS: Ninety-seven percent (73/75) of CNB specimens were considered adequate for a specific diagnosis by the histopathology staff. Diagnostic yield was 97% (95% confidence interval, 91-99%) (72/74) (number of correct diagnoses obtained at CNB / number of definitive diagnoses). There were 61 malignant lesions and 11 benign lesions. There was no false-negative result when CNB was considered adequate for a specific diagnosis by the histopathology staff. Pneumothorax occurred in 19% (12/63 intrapulmonary lesions). One patient required placement of a chest tube. Minor postbiopsy hemoptysis occurred and resolved spontaneously in 11% (7/63) of patients. CONCLUSION: Percutaneous CT-guided coaxial multisampling large CNB of suspected thoracic lesions, in a mainly cancer-based population, is an accurate procedure for a specific histologic diagnosis and has a low rate of complications. PMID- 16247153 TI - Radiofrequency ablation in a porcine lung model: correlation between CT and histopathologic findings. AB - OBJECTIVE: The objective of our study was to investigate the time course changes of the ablated lesion after radiofrequency ablation in the porcine lung and the correlation between CT and histopathologic findings. CONCLUSION: Ground-glass attenuation on CT led to overestimation of the size of necrotic lesions. The layered structural findings on CT were consistent with the histopathologic findings. Although CT findings reflect the histopathologic findings, attention should be paid to the dissociation of ablated lesions and high-density areas in clinical interpretation of CT images. PMID- 16247154 TI - Tumor lysis syndrome after radiofrequency ablation of hepatocellular carcinoma. PMID- 16247155 TI - Benign and malignant mucocele-like tumors of the breast: mammographic and sonographic appearances. AB - OBJECTIVE: The purpose of this study was to analyze and compare the mammographic and sonographic appearances of benign and malignant mucocele-like tumors. CONCLUSION: The mammographic appearance of mucocele-like tumor of the breast is characterized as pleomorphic calcifications, often increasing in number. Microcalcifications in malignant mucocele-like tumors extended over a wider area than those in benign mucocele-like tumors. Sonography often shows cysts with calcified or noncalcified mural nodules. PMID- 16247156 TI - Differentiation of phyllodes breast tumors from fibroadenomas on MRI. AB - OBJECTIVE: The purpose of our study was to evaluate the MRI appearance of phyllodes breast tumors and to differentiate them from fibroadenomas. MATERIALS AND METHODS: MR images were obtained on a 1.5-T imager. T1- and T2-weighted sequences and dynamic 2D fast-field echo T1-weighted sequences were performed. MR images of 23 patients with 24 phyllodes breast tumors (one malignant, 23 benign) were analyzed with respect to morphology and contrast enhancement. The tumors were compared with the MRI appearance of 81 fibroadenomas of 75 patients. RESULTS: Well-defined margins were seen in 87.5% of the phyllodes tumors and 70.4% of the fibroadenomas, and a round or lobulated shape in 100% and 90.1%, respectively. A heterogeneous internal structure was observed in 70.8% of phyllodes tumors and in 49.4% of fibroadenomas. Nonenhancing internal septations were found in 45.8% of phyllodes tumors and 27.2% of fibroadenomas. A significantly greater increase in signal was seen on T2-weighted images in the tissue surrounding phyllodes tumors (21%) compared with fibroadenomas (1.2%). Most of both lesions appeared with low signal intensity on T1- and T2-weighted images. After the administration of contrast material, 33.3% of phyllodes tumors and 22.2% of fibroadenomas showed a suspicious signal intensity-time course. CONCLUSION: Phyllodes breast tumors and other fibroadenomas cannot be precisely differentiated on breast MRI. Phyllodes tumors have benign morphologic features and contrast enhancement characteristics suggestive of malignancy in 33% of cases. PMID- 16247157 TI - Assessment of fetal swallowing with gray-scale and color Doppler sonography. AB - OBJECTIVE: Our study was focused on the evaluation of fluid dynamics to assess the value of gray-scale and color Doppler sonography for evaluating the development of fetal swallow-related movements from early gestation until birth. MATERIALS AND METHODS: We examined 56 fetuses from weeks 15-39 of gestation. Each fetus was examined throughout four distinct periods of gestation: weeks 15-18, 22 25, 30-34, and 37-39. During the examination, seven gray-scale sonography or color Doppler sonography patterns and their prevalence were considered. RESULTS: Mandibular and/or labial movements (chi2 = 56.4, p < 0.0001) and their rhythmic activity (chi2 = 41.4, p < 0.0001) were seen on gray-scale sonography in an increasing percentage of fetuses as gestational age increased. Doppler findings showed an increase for nose-mouth flow signals (chi2 = 57.6, p < 0.0001), larynx esophagus flow signals (chi2 = 13.2, p = 0.0003), and effective swallowing (chi2 = 36.0, p < 0.0001) as gestational age increased. CONCLUSION: There is a trend in the fetus toward development of increased coordinated movement and more functional nose-mouth flow with increasing gestational age: 32.1% of the 56 fetuses in our series achieved effective swallowing at 37-39 weeks, on the basis of gray-scale and Doppler evaluations. Knowledge of the physiologic mechanism involving swallowing development may allow identification of altered swallow related movements in fetuses with malformations of the digestive tract or with neurologic disorders. PMID- 16247158 TI - MRI calculation of lung volumes to predict outcome in fetuses with genitourinary abnormalities. AB - OBJECTIVE: The purpose of our study was to evaluate MRI total lung volumes (TLV) for predicting outcome in fetuses with genitourinary abnormalities and to compare lung volumes with the presence or absence of oligohydramnios. MATERIALS AND METHODS: Fetuses with genitourinary abnormalities underwent blinded retrospective calculation of TLV. Distribution of the TLV-gestational age ratios for survivors and nonsurvivors were compared using the Wilcoxon's rank sum test. Lung volume calculation was compared with the presence or absence of oligohydramnios. RESULTS: There were 21 survivor and 24 nonsurvivor outcomes based on neonatal discharge. TLV-gestational age ratios were significantly different between the survivor and nonsurvivor groups (p = 0.0001). No apparent difference was seen until after 26 weeks of gestation. TLV-gestational age ratios were equal to the presence or absence of oligohydramnios in predicting outcome after 26 weeks of gestation. CONCLUSION: After 26 weeks' gestation, the prediction of outcome in fetal genitourinary abnormalities using the MRI TLV-gestational age ratio is comparable to the presence or absence of oligohydramnios. PMID- 16247159 TI - Posttransplantation lymphoproliferative disorder in pediatric recipients of solid organ transplants: timing and location of disease. AB - OBJECTIVE: The objective of our study was to correlate the location of radiologic presentation and time to onset of posttransplantation lymphoproliferative disorder (PTLD) with the allograft type received in a population of pediatric heart, lung, liver, kidney, and bone marrow transplant recipients. CONCLUSION: Symptomatic PTLD in children manifests earliest in lung recipients and can involve any organ system. However, PTLD in the thorax is most common after lung transplantation, and PTLD in the abdomen most commonly follows kidney transplantation. PMID- 16247160 TI - MDCT diagnosis of the child with posterior plagiocephaly. AB - OBJECTIVE: In this article, we review the normal anatomy and development of the posterior skull base and describe distinguishing imaging features of the two most common causes of posterior plagiocephaly: posterior deformational plagiocephaly and unilateral lambdoid synostosis. We also describe three unusual cases of posterior plagiocephaly, including asymmetric premature fusion of the anterior and posterior intraoccipital synchondroses, with diagnoses enabled by volume reformatted MDCT. CONCLUSION: Three-dimensional reformatted MDCT enables accurate diagnosis of common and rare causes of posterior plagiocephaly in children. PMID- 16247161 TI - Desmoplastic small round cell tumor of the kidney in a pediatric patient: sonographic and multiphase CT findings. PMID- 16247162 TI - Chronic granulomatous lesions after thyroidectomy: imaging findings. AB - OBJECTIVE: The purpose of this report is to describe the imaging appearance of granulomatous inflammation in the neck presenting as a late complication in patients who have undergone thyroidectomy for differentiated thyroid carcinoma. CONCLUSION: Granulomatous inflammation can occur as a palpable mass in the operative bed of asymptomatic patients who have undergone thyroidectomy for thyroid carcinoma. The diagnosis may be suggested when the lesion shows the sonographic appearance of a poorly defined hypoechoic lesion or lesions with a central echogenic nonshadowing focus, often within the sternocleidomastoid muscle. These lesions may appear as complex cystic masses on CT and MRI and may have increased activity on PET. Percutaneous biopsy can establish the diagnosis of an inflammatory lesion and can exclude underlying active infection and malignancy. PMID- 16247163 TI - Unusual anterior neck mass visible only during Valsalva's maneuver in a child. PMID- 16247164 TI - Dimpled appearance of a posterior communicating artery saccule: an angiographic indicator of arterial infundibula. PMID- 16247165 TI - The application of the Six Sigma program for the quality management of the PACS. AB - OBJECTIVE: We implemented a Six Sigma-based quality management program for the PACS to improve the quality of and lessen the necessary resources for its management. CONCLUSION: With the Six Sigma-based PACS quality management program, we were able to reduce resource requirements while maintaining quality. PMID- 16247167 TI - CT evaluation of gastrointestinal stromal tumors treated with imatinib mesylate. PMID- 16247168 TI - Lymphoplasmacytic sclerosing pancreatitis. PMID- 16247169 TI - Moving field of oncologic diagnosis: another reason for going beyond the picture. PMID- 16247172 TI - CT-guided jejunostomy tube insertion. PMID- 16247171 TI - Diagnosis of tailgut cysts. PMID- 16247173 TI - Using movie clips in PowerPoint presentations: part 2, movie editing. AB - OBJECTIVE: The purpose of this article is to describe a simple method for editing movie clips for PowerPoint presentations. CONCLUSION: We describe a simple method for editing movie clips using Adobe Photoshop, a common software utility used by many radiologists for preparing digital images. PMID- 16247174 TI - On the AJR viewbox. Segmental septation of the inferior vena cava. PMID- 16247175 TI - On the AJR viewbox. MR findings of calcaneal intraosseous lipoma with hemorrhage. PMID- 16247176 TI - On the AJR viewbox. Lymph node-like lesion of the neck after pharyngolaryngectomy. PMID- 16247177 TI - TFAR19 gene changes the biophysical properties of murine erythroleukemia cells. AB - TFAR19 is a novel apoptosis-related gene and can accelerate cell apoptosis in the presence of apoptosis inducements. Here, we studied the effects of TFAR19 on some biophysical properties of mouse erythroleukemia (MEL) cells and their molecular and structural basis. After transfected with TFAR19 and apoptosis inducement, MEL revealed a high cell membrane fluidity, a decrease in resynthesis of phospholipids, an increase in the proteins/nucleic acids ratio, a relatively orderly cytoskeleton network, an impaired deformability, a low integrin aM expression, and a decrease in adhesion to endothelial cells. These findings suggest the potential of TFAR19 for antitumor cell migration, and thus for antitumor gene therapy. PMID- 16247180 TI - Controlled autoimmunity in CNS maintenance and repair: naturally occurring CD4+CD25+ regulatory T-Cells at the crossroads of health and disease. AB - T-cells directed to self-antigens ("autoimmune" T-cells) have traditionally been perceived as tending to attack the body's own tissues, and likely to exert their destructive effects unless they undergo deletion in the thymus during ontogeny. Naturally occurring CD4+CD25+ regulatory T-cells were viewed as thymus-derived cells that constitutively suppress any autoimmune T-cells that escaped thymic deletion. Studies in recent years suggest, however, that some autoimmune T-cells are necessary, at least in the central nervous system for neural maintenance and repair, possibly in part by rendering the resident microglia capable of fighting off adverse conditions, as well as for neural maintenance and repair. In line with this notion, the regulatory T-cells are thought to allow autoimmunity to exist in healthy individuals without causing an autoimmune disease. This proposed immune scenario and its implications for therapy are discussed. PMID- 16247179 TI - T-cell cytokines in injury-induced neural damage and repair. AB - T-cell cytokines are involved in beneficial immune responses, pathological autoimmunity, and tissue inflammation. In this review, we focus on the role of interferon-gamma, interleukin (IL)-2, IL-4, IL-6, and IL-10 in autoimmune diseases such as multiple sclerosis, and primarily "nonimmune" injury of the central nervous system (CNS), in particular focal ischemia and trauma. Resident CNS cells such as microglia and astroglia are additional, and on some occasions major, cellular sources of T-cell cytokines in CNS disease. Collectively, they mediate harmful as well as beneficial functions that depend on the dynamics, cellular source, and compartmental site of their release, the pathophysiological context, and the presence of coexpressed factors. Furthermore, direct neurotoxic and neuroprotective effects of cytokines are evident that are independent from their immunoregulating properties. Whereas these complex interactions are only beginning to be understood, T-cell cytokines nevertheless hold promise as therapeutic targets in a variety of neurological disease conditions. PMID- 16247181 TI - T-cells in neuronal injury and repair: semaphorins and related T-cell signals. AB - There are many parallels between the hematopoietic and the nervous systems in terms of mechanisms regulating their development and functions. In neuroinflammatory diseases, interaction between the immune and nervous systems through shared molecules is suspected to trigger an inappropriate crosstalk and lead to demyelination and axonal loss. Here we focus on semaphorins and their functions in the nervous and immune systems and point out the deleterious effect of an immune semaphorin, semaphorin 4D (Sema4D)/CD100, on oligodendrocyte integrity and survival. We propose immune semaphorins as new candidates involved in the pathogenic mechanisms of neuroinflammatory diseases, promoting demyelination, and impairing neuroregeneration. PMID- 16247184 TI - T-cells in human encephalitis. AB - Encephalitis literally means inflammation of the brain. In general, this inflammation can result from a viral or bacterial infection in the brain itself or alternatively from a secondary autoimmune reaction against an infection or a tumor in the rest of the body. Besides this, encephalitis is present in (believed autoimmune) diseases with unknown etiology, such as multiple sclerosis or Rasmussen encephalitis (RE). This article summarizes the existing data on the role of T-cells in the pathogenesis of three types of human encephalitis: RE, paraneoplastic encephalomyelitis, and virus encephalitis. In all of them, T-cells play a major role in disease pathogenesis, mainly mediated by major histocompatiblity complex class I-restricted CD8+ T-lymphocytes. PMID- 16247182 TI - Failed central nervous system regeneration: a downside of immune privilege? AB - Immunity is required to eliminate dangerous or degenerated material and to support regeneration, but also causes significant parenchymal damage. In the eye and the brain, in which cornea and lens poorly regenerate and neurons are hardly replaceable, early transplantation experiments demonstrated remarkable tolerance to various grafts. This "immunologically privileged status" (Billingham and Boswell, 1953) may reflect evolutionary pressure to downmodulate certain actions of immune cells within particularly vulnerable tissues. As an example, tolerating certain "neurotrophic" viruses may often be a more successful strategy for survival than the elimination of all infected neurons. While several constitutive and inducible signals maintaining or re-establishing immune tolerance within the brain have been identified, it has also become evident that the resulting anti inflammatory environment limits certain beneficial effects of neuroinflammation such as neurotrophin secretion or glutamate buffering by T-cells and the clearance of growth-inhibiting myelin or amyloid. Following spinal cord injury, the costs and benefits of neuroinflammation seem to come close because enhancing as well as suppressing innate or adaptive immunity caused amelioration and aggravation of functional regeneration in similar experiments. Evaluating such balances has also begun in (animal models of) Alzheimer's disease, central nervous system trauma, and stroke, and the appreciation of the beneficial side of neuroinflammation has caused a rethinking of the ill-defined use of immune suppressants. As dual roles for individual molecules have been recognized (Merrill and Benveniste, 1996), we are uncovering an already fine-tuned system, but the challenge remains to further support beneficial immune cascades without causing additional damage, and vice versa. PMID- 16247185 TI - T-cells in Alzheimer's disease. AB - Alzheimer's disease (AD) is the most common dementing illness and is pathologically characterized by deposition of the 40-42 amino acid peptide, amyloid-beta (Abeta), as senile plaques. It is well documented that brain inflammatory mechanisms mediated by reactive glia are activated in response to Abeta plaques. A number of reports further suggest that T-cells are activated in AD patients, and that these cells exist both in the periphery and as infiltrates in the brain. We explore the potential role of T-cells in the AD process, a controversial area, by reviewing reports that show disturbed activation profiles and/or altered numbers of various subsets of T-cells in the circulation as well as in the AD brain parenchyma and in cerebral amyloid angiopathy. We also discuss the recent Abeta immunotherapy approach vis-a-vis the activated, autoaggressive T cell infiltrates that contributed to aseptic meningoencephalitis in a small percentage of patients, and present possible alternative approaches that may be both efficacious and safe. Finally, we explore the use of mouse models of AD as a system within which to definitively test the possible contribution of T-cells to AD pathogenesis. PMID- 16247183 TI - Stroke and T-cells. AB - The microvasculature of the brain region affected by a stroke assumes an inflammatory phenotype that is characterized by endothelial cell activation and barrier dysfunction and the recruitment of adherent leukocytes. Although most attention has been devoted to the possible role of neutrophils in the tissue responses to ischemic stroke there is evidence that T-lymphocytes also accumulate in the postischemic brain. Although comparable detailed analyses of lymphocyte involvement in ischemic brain injury have not been performed, emerging findings suggest a role for T-cells in the pathogenesis of ischemic stroke. The recruitment of T-cells to the site of brain injury is critically dependent on the coordinated expression of adhesion molecules on the activated capillary endothelium. Whether the recruited lymphocytes are acting directly on brain tissue or indirectly through activation of other circulating blood cells and/or extravascular cells remain unclear. Cytotoxic CD8+ T-cells may induce brain injury through molecules released from their cytotoxic granules. CD4+ T-helper 1 (TH1) cells, which secrete proinflammatory cytokines, including interleukin-2 (IL 2), IL-12, interferon-gamma, and tumor necrosis factor-alpha, may play a key role in the pathogenesis of stroke, whereas CD4+TH2 cells may play a protective role through anti-inflammatory cytokines such as IL-4, IL-5, IL-10, and IL-13. T-cells should be considered as therapeutic targets for ischemic stroke. However, because infection is a leading cause of mortality in the postacute phase of ischemic stroke, and considering anti-inflammatory role of CD4+TH2, treatment targeting T cells should be carefully designed to reduce deleterious and enhance protective actions of T-cells. PMID- 16247187 TI - Prostaglandins and ulcer healing. AB - Exogenous prostaglandins (PG) applied in small gastroprotective doses fail to affect healing of gastro-duodenal ulcers but accelerate the healing when used in larger gastric inhibitory doses that appear to enhance COX-2 expression and PGE(2) generation in the ulcer area. COX-1 and COX-inhibitors delay ulcer healing, particularly when both COX isoforms are suppressed such e.g. by indomethacin. Dexamethasone, that decreases the expression of COX-2 and mucosal generation of PGE(2), delays ulcer healing that can be reversed by the addition of small dose of exogenous PGE(2). Proton pump inhibitors (PPI) such as omeprazole and PGE analogs, accelerate ulcer healing mainly due to potent inhibition of gastric acid secretion, but they also augment the COX-2 expression and enzyme activity in the ulcerated mucosa. Endogenous PG generated at ulcer margin appear to be involved in ulcer healing promoted by growth factors and gut hormones such as gastrin or CCK and melatonin acting, at least in part, through increase of induction of COX-2 and local release of PGE(2) in the ulcer area. The ulcer healing activity of growth factors (e.g. EGF, TGF alpha, HGF) and certain gut hormones (gastrin, CCK) as well as melatonin, can be attenuated by treatment with COX-1 or COX-2 inhibitors which suppress the release of PGE(2) but enhance the expression of COX-2. It is concluded that endogenous PG originating mainly from upregulated COX-2 at the ulcer margin play crucial role in ulcer healing by exogenous PG, PPI, growth factors, gut hormones and melatonin, while COX-1 and COX-2 inhibitors delay ulcer healing by suppressing PG generation, and increasing COX-2 expression in the ulcer area. PMID- 16247186 TI - T-Cells and excitotoxicity: HIV-1 and other neurodegenerative disorders. AB - Until recently the central nervous system (CNS) was considered an immune privileged site, however, technological and immunological advances have resulted in the CNS being reclassified as an "immune-specialized site." The immune cells, particularly T-cells, continuously patrol the brain and are involved in neuroimmune responses. As such, any changes in the brain microenvironment could affect the physiological functioning of T-cells. Particularly, neurotransmission- associated abnormalities, such as excitotoxicity associated with hypersecretion of glutamate, could severely affect the neuroimmune function of T-cells. Excitotoxicity is involved in the pathogenesis of a number of neurodegenerative disorders. The specific excitotoxicity triggered by the excitatory amino acid neurotransmitter, glutamate, is considered a key mechanism involved in neuronal death. The inability of brain immune cells to overcome these aberrant changes is an active area of investigation. In the systemic circulation, glutamate is inversely related to the number of CD4+ T-cells; however, the effects of elevated glutamate and glutamate-induced exicitotoxicity on cells homing in the brain are critical for understanding neuropathogenesis of neurodegenerative disorders. PMID- 16247188 TI - Role of prostaglandins in gastroprotection and gastric adaptation. AB - Since Robert discovery that pretreatment with prostaglandin (PG) applied in non antisecretory dose can prevent the injury of gastric mucosa induced by necrotizing agents, much attention was paid to the role of these cyclooxygenase (COX) products in the mechanism of gastric mucosal integrity and ulcer healing. The ability of exogenous PG to attenuate or even completely prevent mucosal damage caused by corrosive substances such as absolute ethanol, hyperosmolar solutions or concentrated bile has been termed "cytoprotection". Increased generation of endogenous PG in the gastric mucosa exposed to the topical contact with "mild irritant" such as 20% ethanol, 1 mM NaCl or 5 mM taurocholate also prevented gastric injury caused by strong irritants via phenomenon of adaptive cytoprotection. Other mediators such as growth factors, nitric oxide (NO) or calcitonin gene related peptide (CGRP) as well as some gut hormones including gastrin and cholecystokinin (CCK), leptin, ghrelin and gastrin-releasing peptide (GRP) have been also found to protect gastric mucosa against the damage induced by corrosive substances. This protective action of gut hormones has been attributed to the release of PG or activation of sensory nerves because it could be abolished by the pretreatment with indomethacin or large neurotoxic dose of capsaicin and restored by the addition of exogenous PGE(2) or CGRP, respectively. Short (5 min) ischemia of the stomach applied before prolonged ischemia reperfusion (I/R) attenuated markedly the gastric lesions produced by this I/R and also prevented the mucosal damage provoked by necrotizing substances. This protection could be abolished by the pretreatment with non-steroidal anti inflammatory drugs (NSAID) and was accompanied by an enhancement of gastric mucosal COX-2 expression and activity. Exposure of gastric mucosa to single insult of acidified aspirin (ASA) causes severe mucosal damage with occurrence of multiple haemorrhagic lesions but with repeated application of ASA, the attenuation of mucosal lesions is observed, despite the profound inhibition of PGE(2) generation. This phenomenon called "gastric adaptation" does not appear to depend upon endogenous biosynthesis of PG but possibly involves enhanced production of growth factors increasing cell proliferation and mucosal regeneration. Unlike short lived gastroprotection by PG, NO, CGRP, mild irritants or short ischemia, gastric adaptation appears to be long-lasting phenomenon accompanied by increased resistance of the adapted mucosa to subsequent damage induced by corrosive agents. PMID- 16247189 TI - Prostaglandins as mediators of COX-2 derived carcinogenesis in gastrointestinal tract. AB - This review was designed to show the role of expression of cyclooxygenase (COX)-1 and COX-2 in the cancerogenesis of esophagus, stomach and colon. Unlike COX-1, which is expressed in the normal esophago-gastro-colonic mucosa, COX-2 was found to be expressed mainly in the pre-cancer changes in the mucosa including Barrett's esophagus, Helicobacter pylori (H. pylori)-induced gastritis and inflammatory changes in colonic mucosa. In Barrett's esophagus, prostaglandins (PGs) derived from upregulated COX-2 contribute to the progression of low-grade to high-grade dysplasia and finally to cancer. In chronic gastritis induced by chronic H. pylori infection, overexpression of COX-2 is probably induced by inflammatory cytokines, growth factors, especially gastrin and reactive oxygen species leading to mutagenesis and subsequent metaplasia, dysplasia and cancer formation. The imbalance between cell proliferation and apoptosis caused mainly by products of COX-2 leads to cancerogenesis. Similarly, in colorectal cancer the overexpression of COX-2, possibly induced by the action of growth promoting factors including progastrin and gastrin and overexpression of survivin contribute to the colorectal cancerogenesis that could be, at least in part, amended by the treatment with specific COX-2 inhibitors. We conclude that: 1) COX 2-derived PGs play a key role in the tumorigenesis in the gastrointestinal tract; 2) The tumor-promoting effect of PGs may be attributed to their ability to stimulate cell proliferation and migration, to inhibit the apoptosis and to increase angiogenesis and invasiveness; 3) In accordance to the proposed major role of COX-2 in cancerogenesis, selective COX-2 inhibitors have been shown in numerous studies to exhibit strong chemopreventive effect on the development of gastrointestinal cancers. PMID- 16247190 TI - Comparison of prostaglandin and cimetidine in protection of isolated gastric glands against indomethacin injury. AB - Prostaglandins can protect the in vivo gastric mucosa against necrosis produced by a variety noxious agents. Cimetidine has also been shown to have protective properties in humans and in some models of experimental injury. Whether prostaglandins or cimetidine may protect gastric mucosal cells directly in the absence of systemic factors remains controversial. In the present study, the potential protective actions of prostaglandin and cimetidine against indomethacin injury were assessed in isolated rat gastric glands. Gastric glands were pre incubated in oxygenated medium with either placebo, 16,16 dimethyl prostaglandin E(2) (dm PGE(2)) or cimetidine and incubated at 37 degrees C in medium containing 0.5 mg/ml of indomethacin for 2, 4 and 6 hrs. Cell injury and protection was assessed by the Fast Green exclusion test (viability test), leakage of lactate dehydrogenase (LDH) into the medium, and by scanning and transmission electron microscopy. In addition, the generation of PGE(2) by the gland cells was determined using RIA assay. Indomethacin by itself significantly reduced the viability of gastric glands, increased LDH release into the medium and produced prominent ultrastructural damage. In contrast to cimetidine, co-incubation of gastric glands with dm PGE(2) added to indomethacin, significantly reduced indomethacin-induced injury, increased the number of viable cells, reduced LDH leakage and diminished the extent of ultrastructural damage. The dose of indomethacin (5 microg/ml) which significantly inhibited the generation of PGE(2) (up to 90% inhibition) had no effect on cell viability nor LDH release. We conclude that 1) exogenous PGE2 exerts a potent protective activity in vitro which is independent on neural, vascular and hormonal factors; 2) inhibition of endogenous PGs may not the primary mechanism in the deleterious action of indomethacin against damage to gastric glandular cells and 3) indomethacin can exert a direct cytotoxic effect on the mucosal cells in gastric glands. PMID- 16247191 TI - Significance of salicylate intolerance in diseases of the lower gastrointestinal tract. AB - Salicylate intolerance is defined as a nonspecific antigen-induced pseudo allergic hypersensitivity reaction which can occur upon contact of an organism with salicylic acid, its derivatives or other related organic or inorganic acids of similar chemical structure. Since the effects of nonsteroidal anti inflammatory drugs (NSAID) intolerance are by no means always severe or life endangering but may just as well present as oligosymptomatic or local disorders (e.g. abdominal pain, diarrhea, we decided to evaluate the characteristics of patients with salicylate intolerance on the basis of gastroenterological case material of Medical Department I of Erlangen University. On the basis of the findings from the Erlangen interdisciplinary data register of chronic inflammatory gastrointestinal disease, the signs and symptoms of NSAID intolerance were found to constitute a diagnosis of great practical import to clinical medicine (allergology, dermatology, immunology, other disorders etc.) including gastroenterology. For approx. 2-7% of all patients with inflammatory bowel syndrome and food allergies this poses a new diagnostic and therapeutic challenge which may concern physicians from any of the disciplines involved. When presented with patients with chronic active disease who are suffering from these symptoms one should, therefore, in future give greater thought to the possibility of salicylate intolerance, all the more as there are meaningful dietetic, diagnostic and therapeutic options available for these persons. PMID- 16247192 TI - Functional eicosanoid test and typing (FET) of peripheral blood cells in eicosanoids related diseases. AB - Monitoring of eicosanoid synthesis in peripheral blood cells has significant potential for improving the diagnosis and therapy of many human diseases. The quantitative relation between concentrations of prostaglandins and leukotrienes is central to the physiologic function of the eicosanoid network. Here we show that this regulation, which we call the functional eicosanoid typing (FET), fluctuates dynamically in individual living blood cells from patients, thereby limiting the accuracy with which concentration circuits of eicosanoids can transfer metabolic information. Using living cells in functional cell testing, we characterised the eicosanoid pattern score (EPS). A novel technique based on binomial errors on lipid mediator partitioning enabled calibration of in vivo biochemical parameters in molecular units. We found that eicosanoid production rates fluctuate over a time scale of about twenty minutes, while intrinsic noise decays rapidly. Thus, biochemical eicosanoid parameters, noise, and slowly varying cellular states together determine the effective FET. These results can form a basis for quantitative modelling of natural eicosanoid circuits in diagnosis of eicosanoid related diseases and design of synthetic ones for the prediction other diseases. PMID- 16247193 TI - Abnormal eicosanoid-pattern of peripheral white-blood cells in gastro-intestinal cancer. AB - COX-inhibitors promote nasal polyps or bronchial asthma in individuals susceptible to an alteration of the pattern of the eicosanoids, especially leukotrienes and prostaglandins. This is associated with an abnormal release of eicosanoids from white blood cells. Since COX-inhibitors also protect from colorectal cancer an analogous association may be suggested. The study was performed to detect abnormal patterns of eicosanoids in white blood cells of patients with intestinal cancer compared to healthy controls. Seventy patients with intestinal cancer (stomach = 5; colon = 25; sigma = 18; rectum = 22) were compared to 62 healthy controls. Blood leukocytes from patients in complete long lasting remission were incubated with diluent, arachidonic acid or acetylsalicylic acid. The synthesis of prostaglandin E(2) and peptido leukotrienes was quantified using competitive enzyme-immuno-assays and calculated for individual eicosanoid patterns. The mean basal and arachidonic- or acetylsalicylic acid-modulated PGE(2) synthesis in patients was significantly higher than in controls (4.8-fold, 9.4-fold, 3.7-fold, respectively) whereas pLT was generally less elevated. We conclude that the eicosanoid-pattern of white blood-cells from patients with intestinal cancer differs significantly from that in healthy individuals. This abnormal cellular metabolism may contribute to the manifestation of cancer and help to detect individuals at risk. PMID- 16247196 TI - A new low-friction ligation system. PMID- 16247194 TI - Shear bond strengths using an indirect technique with different light sources. PMID- 16247197 TI - Bonding impacted teeth without moisture contamination. PMID- 16247198 TI - An alternative method of space closure: the Hycon Device. PMID- 16247199 TI - Correction of incisor position after breakage of a fixed lingual retainer. PMID- 16247200 TI - Modified intrusive mechanics in lingual segmented-arch treatment. PMID- 16247201 TI - Pulmonary thromboembolism associated with air travel in Japan. AB - BACKGROUND: A link between venous thromboembolism (VTE) and air travel has been recently discussed, so the present study aimed to clarify the characteristics of acute pulmonary thromboembolism (PTE) associated with air travel in Japan. METHODS AND RESULTS: The study group comprised 36 patients with acute PTE associated with air travel. Patients' characteristics, clinical risk factors for VTE and coagulation abnormalities were investigated, and compared with a group of PTE not associated with travel. Of the 36 cases, 30 cases were female, 34 were over 40 years of age and all had flown more than 6 h. Two-thirds of all cases did not leave their seats during the flight. Of the patients, 44% had no clinical risk factors for VTE or coagulation abnormalities. Idiopathic PTE (ie, PTE without clinical risk factors or coagulation abnormalities) was significantly more common in the air travel group. CONCLUSION: Acute PTE associated with air travel in Japan is common in elderly women and may occur without any clinical risk factors for VTE or coagulation abnormalities. Remaining seated throughout the flight increases the risk. PMID- 16247203 TI - Calcium antagonists for secondary prevention of myocardial infarction: is there a need to shift from short-acting to long-acting types? AB - BACKGROUND: Although short-acting nifedipine does not prevent myocardial infarction (MI), calcium antagonists with a long half-life may be effective. METHODS AND RESULTS: The present study was a retrospective analysis of the incidences of cardiac events among patients with a healed MI treated with 3 times a-day type nifedipine (half-life 1.8 h; n=617), twice-a-day type nifedipine (half life 4.0 h; n=527) and those not taking calcium antagonists (n=1,593) from 1986 to 1993, and the incidences of those on once-a-day type calcium antagonists (half life 11.0 h; n=903) and those not taking calcium antagonists (n=2,788) from 1994 to 2001. Cardiac events included cardiac death and nonfatal recurrent MI. Single and multivariate analyses using the Cox-Hazard model were performed. From 1986 to 1993 cardiac events occurred in 38 patients with 3-times-a-day nifedipine (6.2%, hazard ratio and 95% confidence interval: 1.45 and 0.93-2.27), in 18 patients with twice-a-day nifedipine (3.4%: 0.68 and 0.39-1.20), 57 patients without calcium antagonists (3.6%). Cardiac events also occurred in 11 patients with once a-day type nifedipine (1.2%: 0.72 and 0.37-1.42) and 48 patients without calcium antagonists (1.7%). Of the once-a-day type calcium antagonists, cardiac events were the lowest (2 patients, 0.6%: 0.32 and 0.08-1.31) in patients with amlodipine (half-life 39.0 h; n=334), which has the longest half-life. None of these drugs were a significant factor in the increase or decrease in the incidence of cardiac events. However, there was a good correlation between the half-life of the calcium antagonist and the hazard ratio for reducing cardiac events. CONCLUSION: Although there was a tendency toward a reduction in cardiac events using calcium antagonists with long half-life, none of these drugs could significantly reduce the incidence of cardiac events in patients with a healed MI. PMID- 16247202 TI - Early defibrillation and circulatory support can provide better long-term outcomes through favorable neurological recovery in patients with out-of-hospital cardiac arrest of cardiac origin. AB - BACKGROUND: Early defibrillation and cardiopulmonary bypass have been postulated to be a promising intervention against out-of-hospital cardiac arrest (OHCA); however, little is known about the long-term prognosis. The effects of early recovery of circulation (ROC) on neurological recovery and the long-term outcome in patients with OHCA were examined. METHODS AND RESULTS: Functional recovery and long-term (22.0+/-15.3 months) outcome were examined in 100 patients with definite diagnosis of OHCA. Spontaneous circulation recovered in 79% of the patients (using on-site counter shock in 20% of the patients). Cardiopulmonary bypass was performed in 38 of the OHCA patients. The total survival and favorable neurological recovery rates were 40% and 25%, respectively. The patients with favorable recovery obtained early ROC (28.2+/-16.0 min). Receiver-operating characteristic analysis showed that a period of less than 35 min for ROC was the optimal period for achieving a favorable recovery, with sensitivity of 68% and specificity of 73%. The patients with a prior history of heart failure or reduced left ventricular ejection fraction exhibited more frequent, exacerbated heart failure and ventricular arrhythmias. CONCLUSIONS: Early ROC using on-site counter shock or cardiopulmonary bypass might result in better long-term outcome in patients with OHCA of cardiac origin. PMID- 16247204 TI - Angiotensin-converting enzyme genotype is not associated with exercise capacity or the training effect of cardiac rehabilitation in patients after acute myocardial infarction. AB - BACKGROUND: The relationship of the genotype for the angiotensin-converting enzyme (ACE) with exercise capacity or training effects has been studied in athletes or healthy persons, but recently the ACE DD genotype was reported to be associated with decreased exercise capacity in patients with congestive heart failure. Therefore, in the present study the association between the ACE genotype and exercise capacity was investigated in patients with acute myocardial infarction (AMI) participating in cardiac rehabilitation (CR) for 3 months. METHODS AND RESULTS: The study population comprised 168 patients stratified as II (n=75), ID (n=67), and DD (n=26) according to ACE genotype. Baseline left ventricular ejection fraction (LVEF) was similar among the genotype groups. In all patients, exercise capacity (peak work rate (PWR) and peak oxygen uptake (PVO 2)) significantly increased after CR. However, no differences were observed in PWR and PVO2 among the genotype groups at baseline or after CR. The results were similar even when analyzed in 60 patients with left ventricular (LV) dysfunction (LVEF <45%). CONCLUSION: The present study suggests that there is no association between ACE I/D polymorphism and exercise capacity in patients after AMI, even with LV dysfunction. Furthermore, ACE genotype may have no influence on the effects of CR after AMI. PMID- 16247205 TI - Nocturnal oxygen desaturation as a predictive risk factor for coronary restenosis after coronary intervention. AB - BACKGROUND: Sleep-disordered breathing may affect coronary artery disease, so patients treated by percutaneous transluminal coronary angioplasty were studied to investigate the relationship between nocturnal oxygen desaturation (NOD) and the loss index. METHODS AND RESULTS: The nocturnal oxygen desaturation index (ODI) was determined by means of a pulse oximeter in 35 patients with coronary artery disease treated by stent placement. The patients were divided into 3 groups according to ODI: <5 events/h (group N, n=6), between 5 and 14 events/h (group A, n=19), and >or=15 events/h (group B, n=10). The relationship between the ODI and loss index was examined in each group. The loss index, 0.27+/-0.23 in group N, 0.30+/-0.27 in group A, and 0.70+/-0.32 in group B, differed significantly between groups N and B (p=0.007). In all patients, the loss index showed a significant positive correlation with the ODI (R=0.585, p=0.002). CONCLUSIONS: These findings suggest that NOD may be an important contributor to coronary restenosis in patients treated with stent placement. PMID- 16247206 TI - Association of case volume with mortality of chinese patients after coronary artery bypass grafting: Taiwan experience. AB - BACKGROUND: An inverse relationship between surgeon volume or hospital volume and mortality has been reported in Western countries, but seldom in Asia. METHODS AND RESULTS: The data of 4,724 patients with coronary artery disease who underwent coronary artery bypass graft (CABG) surgery in Taiwan between 1(st) January 2000 and 31(st) December 2001 were analyzed in this prospective cohort study. Overall, 3.45% of patients died in-hospital (IH), and 6.48% patients died within 30 days after discharge (AD30); 85.0% of patients in the AD30 group died at home within 1 day of discharge because of a "cultural preference for dying at home". After adjustment by stepwise logistic regression for age, sex, cardiac function, co morbidity and in-hospital complications, higher provider volume was still associated with lower mortality rates for CABG, especially higher surgeon volume. Because IH mortality can be affected by different culture, the IH plus AD30 mortality rate is more appropriate as a proxy to reflect the mortality of a specific procedure. CONCLUSIONS: The relationship of higher-volume hospitals or surgeons with lower mortality rate for patients undergoing CABG is a general phenomenon in Western and Chinese societies. However, the combination of the AD30 and IH mortality rates has to be considered when investigating procedural mortality rates in Chinese society. PMID- 16247207 TI - Need for documentation of guidelines for coronary artery spasm: an investigation by questionnaire in Japan. AB - BACKGROUND: Because there are no guidelines concerning coronary spasm in Japan, the present study examined the current status of the spasm provocation test. METHODS AND RESULTS: Questionnaires concerning the number of cases of coronary angiography, percutaneous coronary intervention, and invasive/non-invasive spasm provocation tests over 3 years (2001-2003) and the status of spasm provocation tests were sent to members of the Japanese Circulation Society in 120 cardiology hospitals in the Chugoku and Shikoku areas. Completed surveys were returned from 45 hospitals, giving a collection rate of 38%. As a spasm provocation agent, acetylcholine tests were performed in 29 hospitals, and ergonovine tests in 18 hospitals. Non-invasive spasm provocation tests were performed in only 9 hospitals (20%). In total, 5,267 patients underwent acetylcholine spasm provocation test (2,387 patients) or ergonovine spasm provocation test (2,880 patients) and vasospastic angina was diagnosed in 1,663 (2.4%) patients. Invasive spasm provocation tests were performed in 7.8% of patients with diagnostic catheterization and the spasm-positive rate was 31.6%. The difference among hospitals concerning the number of invasive spasm provocation tests was remarkable, and the angiographic spasm-positive standard and acetylcholine/ergonovine dose varied among the hospitals. CONCLUSIONS: Guidelines on coronary spasm in Japan are essential to overcome the current differences between institutions. PMID- 16247209 TI - Evaluation by multislice computed tomography of atherosclerotic coronary artery plaques in non-culprit, remote coronary arteries of patients with acute coronary syndrome. AB - BACKGROUND: Patients with acute coronary syndrome (ACS) frequently have vulnerable plaques in the remote coronary arteries, suggesting that ACS is part of the pan-coronary process. In the present study the computed tomography (CT) plaque density in non-culprit atherosclerotic coronary artery lesions was evaluated by multislice computed tomography (MSCT) in patients with ACS and non ACS. METHODS AND RESULTS: MSCT was performed in 21 patients with ACS and 53 patients with non-ACS: 16 of the 21 ACS patients (76%) and 30 of the non-ACS 53 patients (57 %) had non-calcified plaques in the non-culprit coronary arteries (p=0.18). CT-low-density plaques (CT density <68 Hounsfield units (HU)) were more frequent in the ACS group (13/16 patients, 81%) than in the non-ACS group (13/30 patients, 43%, p=0.03). In addition, the CT density of the non-culprit lesion was significantly lower in patients with ACS than in those with non-ACS (44.1+/-22.9 and 77.3+/-33.7 HU, respectively). CONCLUSION: Patients with ACS more frequently had CT-low-density plaques in the non-culprit, remote arteries than those with non-ACS, which suggests that ACS treatment should focus not only on stabilizing the culprit lesion but also on systemic stabilization of non-culprit lesions. PMID- 16247208 TI - Comparison of post-exercise and post-vasodilator stress myocardial stunning as assessed by electrocardiogram-gated single-photon emission computed tomography. AB - BACKGROUND: Exercise gated single-photon emission computed tomography (SPECT) using technetium-99m (99mTc)-sestamibi evaluates both myocardial perfusion during stress and wall motion >30 min after the stress, which has the potential to assess not only exercise-induced myocardial ischemia but also the development of myocardial stunning. METHODS AND RESULTS: To evaluate the incidence of post stress myocardial stunning, as well as comparing the effects of different stress methods on the development of stunning, 179 consecutive patients with known or suspected coronary artery disease (CAD) underwent (99m)Tc-sestamibi SPECT with either exercise (n=135) or adenosine triphosphate disodium (ATP) (n=44). Electrocardiogram-gated SPECT images were acquired >30 min after the stress and again 4 h later, and perfusion and wall motion were evaluated. Post-stress myocardial stunning occurred in 24 patients (13%): 22 after exercise and 2 after ATP stress. The magnitude of the transient wall motion abnormality after exercise was greater in patients with severe ischemia, compared with those with mild-to moderate ischemia (p<0.0001). By contrast, with ATP stress, the magnitude of the transient wall motion abnormality was similar, regardless of the severity of perfusion abnormality. Furthermore, a significant correlation between summed difference score and transient wall motion abnormality was found after exercise (r=0.68, p<0.0001). With ATP, however, no such correlation was observed (r=0.28, p=NS). CONCLUSIONS: Using 99mTc-sestamibi gated SPECT, myocardial stunning is frequently observed after exercise and correlates with the severity of myocardial ischemia, but this does not occur with ATP, which is regarded as a specific marker for severe CAD. PMID- 16247210 TI - Evaluation of cardiac tumors in children by electron-beam computed tomography: rhabdomyoma and fibroma. AB - BACKGROUND: The most common cardiac tumors in young children are rhabdomyoma and fibroma, and prognosis depends on the tumor's characteristics. In the present study electron-beam computed tomography (EBT) was used to evaluate tumor characteristics and myocardial perfusion in 6 children with cardiac tumors. METHODS AND RESULTS: Five children had rhabdomyomas and 1 had a fibroma. In the precontrast study, the computed tomography numbers of rhabdomyomas were higher and those of the fibroma were slightly lower than those of the normal myocardium. In the early postcontrast study, the size, location, shape and number of the tumor were clearly detected, and in the late postcontrast study, the fibrous characteristics of the tumor were seen. The fibroma contained calcification and 2 rhabdomyomas contained a fat density spot. In the cine-mode study, both the tumor and ventricular wall motion could be simultaneously evaluated. There was a filling defect of the left ventricular myocardium on the early postcontrast scan of 1 child with a rhabdomyoma and dyskinetic wall motion at the same region in the cine-mode study. CONCLUSION: EBT was useful for evaluating tumor characteristics in children. PMID- 16247211 TI - Multicenter survey on the validity of the CD-ROM guideline for antiarrhythmic drug therapy produced by the Japanese Circulation Society and the Japanese Society on Electrocardiology: preliminary report of the survey of the Japanese guideline for Arrhythmia Management By Individual Therapy (J-GAMBIT). AB - BACKGROUND: A multicenter investigational survey (Japanese Guideline for Arrhythmia Management By Individual Therapy) was conducted to evaluate the validity of using CD-ROM guidelines vs physician choice for the selection of antiarrhythmic drugs. METHODS AND RESULTS: Patients with paroxysmal atrial fibrillation (PAF, n=274) or premature ventricular contractions (PVC, n=216) were enrolled. The rate of concordance for drug selection between the treating physician and the CD-ROM was 216 of 274 patients (78.8%) with PAF. Of these, 168 (61.3%) were concordant for first-line agents and the remaining 48 (17.5%) were concordant for second-line agents. The concordance for the treatment of PVC was 154/216 cases (71.3%). Of these, 106 (49.1%) were concordant for first-line agents and the remaining 48 (22.2%) were concordant for second-line agents. Nonconcordance for PAF therapy was more likely to occur for patients with underlying heart disease (p<0.05), depressed cardiac function (p<0.001), and with more frequent ECG abnormalities and renal dysfunction. These differences were not seen in patients with PVC. CONCLUSION: The CD-ROM guidelines appear to be valid in the selection of antiarrhythmic drugs for both PAF and PVC, but their usefulness is influenced by the patient's clinical characteristics. PMID- 16247212 TI - Monophasic action potential duration at the crista terminalis in patients with sinus node disease. AB - BACKGROUND: The repolarization properties of the crista terminalis (CT) cells have not been elucidated in patients with sinus node disease (SND). In the present study a new technique of recording the monophasic action potential (MAP) at the CT was used to examine the repolarization of the right atrium (RA) in SND patients. METHODS AND RESULTS: Symptomatic SND (n=13) patients and age-, sex matched control patients (n=13) were tested. The MAP duration (MAPD) at a basic cycle length of 600 ms was recorded at the CT in the superior vena cava - RA junction and at the middle - anterior RA with the effective refractory period (ERP) at the high RA. In 6 controls and 4 SND patients, the effect of adenosine triphosphate on the MAPD was examined. The MAPD at the CT exceeded that at the middle - anterior RA in both groups. The MAPD at the CT in the SND group was significantly prolonged compared with the control group (CT: 358+/-39 ms vs 289+/ 43 ms). Between the SND and control groups, the MAPD at the middle - anterior RA (278+/-36 ms vs 265+/-39 ms) and ERP (294+/-42 ms vs 266+/-41 ms) did not differ. Both the corrected-sinus node recovery time and sinoatrial conduction time were better correlated with the MAPD at the CT than the MAPD at the middle - anterior RA and ERP. Adenosine triphosphate shortened the MAPD, which was augmented at the CT in the SND patients. CONCLUSION: A novel method of estimating the MAP at the CT revealed the characteristics of atrial repolarization in SND patients. PMID- 16247213 TI - Discordant repolarization alternans-induced atrial fibrillation is suppressed by verapamil. AB - BACKGROUND: Ventricular alternans of repolarization produces serious ventricular arrhythmias in experimental models. The present study investigated the role of alternans of atrial repolarization in patients with atrial fibrillation (AF). METHODS AND RESULTS: Electrophysiological studies were performed in 19 patients without structural heart disease. Monophasic action potentials (MAP) were recorded with 2 Franz catheters during steady state pacing, starting at a cycle length (CL) of 400 ms with subsequent decrements of 10 ms. Duration from the onset of upstroke to 90% repolarization of the MAP were measured. If discordant alternans (DA) was present during pacing, verapamil was administrated, and MAP measurements were repeated. Rapid pacing resulted in concordant alternans to DA in 13 of 19 (68%) patients. AF was initiated after the induction of DA in 8 of 13 patients (p=0.012). Verapamil treatment resulted in a significant decrease in the longest pacing CL at which DA was induced (207+/-19 vs 178+/-17 ms, p<0.0001). CONCLUSIONS: Rapid atrial pacing induced DA and was associated with initiation of AF. Furthermore, induction of DA was suppressed by verapamil. Reducing the spatiotemporal repolarization heterogeneity may be how the calcium-channel blockade prevents initiation of AF. PMID- 16247214 TI - Prospective study of the changes in left ventricular mass and geometry patterns in hypertensive patients During 5 years of follow-up. AB - BACKGROUND: Left ventricular hypertrophy (LVH), as well as the geometry pattern of the left ventricle, is believed to be an independent risk factor for hypertension. The present study investigated the changes in left ventricular mass, diastolic function and geometry in hypertensive patients in a prospective 5 year follow-up in conjunction with an evaluation of the regularity and effectiveness of treatment. METHODS AND RESULTS: One hundred hypertensive patients older than 18 years were examined according to the study protocol, which included registration of weight, height, vital signs, and echocardiography. After 5 years a repeat examination was performed. Patients were divided into 3 groups according to blood pressure (BP) control: group 1 (n=32), no regular medication; group 2 (n=44), regular treatment but no target BP levels; group 3 (n=14), regular effective treatment. In group 1 an increase in LVH and worsening of diastolic function were observed; in group 2 LVH and isovolumetric relaxation time remained unchanged, while the early peak velocity/atrial peak velocity ratio decreased; in group 3 there was a significant decrease in LVH. The geometry pattern only changed in 21 (23%) patients. CONCLUSIONS: LVH can be successfully reversed in only hypertensive with adequate BP control. The remodeling pattern appears to be a stable characteristic of the patient and transformation of one pattern into another is infrequent. PMID- 16247215 TI - Low-renin (volume dependent) mild-hypertensive patients have impaired flow mediated and glyceryl-trinitrate stimulated vascular reactivity. AB - BACKGROUND: Low-renin (volume-dependent) hypertension represents 25-30% of all cases of primary hypertension. Endothelial dysfunction and vascular remodeling are associated with hypertension but their relevance to volume-dependent hypertension (VDH) is not yet known. To evaluate this, flow-mediated dilation (FMD) of the brachial artery and the carotid intima-media thickness in the distal common carotid artery were measured and compared between renin-dependent mild hypertensive patients (RDH) and controls. METHOD AND RESULTS: The study group comprised 40 mild-hypertensive patients and 25 controls. Plasma renin activity (PRA), plasma aldosterone concentration, angiotensin II and nitrite/nitrate plasma levels were measured. According to PRA, subjects were classified as VDH (<0.6 ng . ml (-1) . h(-1)), or RDH (>0.6 ng . ml(-1) . h (-1)). Vascular function was evaluated by FMD before and after reactive hyperemia (RH) and glyceryl-trinitrate (GTN) administration. FMD in response to RH and GTN in the VDH group when compared with RDH group was 10.2+/-2.8% vs 13.3+/-3.6% (p=0.01); and 16.0+/-3.5% vs 19.9+/-4.5% (p=0.01), respectively. CONCLUSION: This study showed impaired FMD and reduced GTN response in mildly hypertensive patients with low-renin plasma levels. PMID- 16247216 TI - Portopulmonary hypertension: hemodynamics, pulmonary angiography, and configuration of the heart. AB - BACKGROUND: The goal of the present study was to examine the cardiac configuration and pulmonary vascular changes in patients with portopulmonary hypertension (PPHTN) and compare them with those of idiopathic pulmonary arterial hypertension (IPAH). METHODS AND RESULTS: The subjects were 10 patients with PPHTN and 18 with IPAH. In PPHTN, the increases in the right ventricular end diastolic volume index (89+/-19 vs 128+/-50 ml/m2; p=0.04), right end-systolic volume index (50+/-19 vs 95+/-47 ml/m 2; p=0.02) and right ventricular mass index (47+/-18 g/m2 vs 79+/-31; p=0.04) were low compared with IPAH. The decrease in the right ventricular ejection fraction was also low in PPHTN (45+/-10 vs 28+/ 13%; p=0.01). The degree of sparse arborization and abrupt narrowing on wedged pulmonary angiography was moderate in PPHTN compared with IPAH. In PPHTN, the proximal pulmonary arteries were dilated near the segmental arteries, which were narrow in IPAH. CONCLUSION: Changes in the configuration of the heart were moderate in PPHTN compared with those in IPAH. The degree of sparse arborization and abrupt narrowing were also moderate in PPHTN. PMID- 16247217 TI - Long-term effect of spironolactone on cardiac structure as assessed by analysis of ultrasonic radio-frequency signals in patients with ventricular hypertrophy. AB - BACKGROUND: An effect of aldosterone on ventricular fibrosis has been demonstrated in animals, but remains unclear in human patients. This study aimed to investigate (1) the relationship between left ventricular (LV) fibrosis and myocardial ultrasonic texture as assessed with myocardial radio-frequency (RF) signals analyzed from the viewpoint of their waveform with chaos theory in animals and (2) serial changes in myocardial ultrasonic texture following long term aldosterone blockade in patients with LV hypertrophy. METHODS AND RESULTS: In an animal study, Sprague-Dawley rats were divided into 2 groups with and without adriamycin administration, and the relationship between the RF signals and LV fibrosis was assessed. In a clinical study, effects of 12-month administration of spironolactone were assessed in patients with LV hypertrophy. The animal study revealed that the correlation dimension (CD) calculated from the RF signals inversely correlated with the area of fibrosis. The clinical study demonstrated an increase in CD following 6-month administration of spironolactone. The changes in CD positively correlated with those in the serum carboxy-terminal telopeptide of collagen type I. CONCLUSION: Myocardial RF signals analyzed with chaos theory reflect the severity of LV fibrosis. Aldosterone blockade may alter myocardial ultrasonic texture with regression of LV fibrosis, at least partly through enhanced collagen degradation. PMID- 16247218 TI - Regulation of arginine vasopressin in the human heart. AB - BACKGROUND: The role of arginine vasopressin (AVP) in the heart has yet to be determined. The present study was designed to examine whether AVP is regulated in the human heart. METHODS AND RESULTS: The subjects were 93 patients who underwent coronary angiography and left ventriculography. Blood samples were collected at the aortic root (AO) and the coronary sinus (CS) to measure the plasma levels of AVP. The patients who showed increases in AVP levels at the CS and AO were assigned to the increased AVP group and those who showed no change or a decrease were assigned to the non-increased AVP group. Cardiac function was compared between these 2 groups. There was a significant difference (p<0.0234) in left ventricular end-diastolic volume index between the increased AVP group (125.5 +/ 53.4 ml/m2) and the non-increased AVP group (102.2+/-30.6 ml/m2). There was also a significant difference (p<0.0137) in left ventricular stroke volume index between the increased AVP group (66.6+/-23.2 ml/m2) and the non-increased AVP group (54.4+/-18.6 ml/m2). CONCLUSION: These results suggest that both the production of AVP and synthesis with its receptors may be enhanced at regional sites of the human heart in the volume load. PMID- 16247219 TI - Subtype switching of L-Type Ca 2+ channel from Cav1.3 to Cav1.2 in embryonic murine ventricle. AB - BACKGROUND: Embryonic hearts exhibit spontaneous electrical activity, which depends on Ca2+ influx through L-type Ca2+ channels. In this study the expression of the L-type Ca2+ channel alpha1 subunit gene in the developing mouse heart was investigated. METHODS AND RESULTS: Mouse cardiac ventricles 9.5 days post coitum (dpc), 18 dpc and adult were used. At 9.5 dpc the level of Cav1.3 mRNA was higher than that of Cav1.2 mRNA. With development, Cav1.2 mRNA increased and Cav1.3 mRNA decreased. Analysis of Cav1.3 splicing variants showed that Cav1.3(1b) mRNA was expressed at a higher density than Cav1.3(1a) mRNA. Cav1.3 protein was detected only at 9.5 dpc, whereas Cav1.2 protein was expressed from 9.5 dpc and its expression increased with development. L-type Ca2+ currents were prominent at 9.5 dpc. The Ca2+ current amplitude at 9.5 dpc was comparable to that at 18 dpc, and was larger in adults than at the embryonic stage. L-type Ca2+ current at 9.5 dpc was activated and/or inactivated at more negative membrane potentials than at 18 dpc or adult. L-type Ca2+ channels at 9.5 dpc were less sensitive to inhibition by nisoldipine than at adult. CONCLUSIONS: The Cav1.3 channel is functionally expressed in early embryonic mouse ventricular myocytes and potentially underlies ventricular automaticity. PMID- 16247220 TI - SOCS1/JAB likely mediates the protective effect of cardiotrophin-1 against lipopolysaccharide-induced left ventricular dysfunction in vivo. AB - BACKGROUND: Suppressor of cytokine signaling 1 (SOCS1) is a negative regulator of cytokine signaling whose expression is induced in the rat heart by cardiotrophin 1 (CT-1). Sepsis-induced myocardial depression results from the expression of inducible nitric oxide synthase (iNOS) evoked by inflammatory cytokines. METHODS AND RESULTS: The effect of CT-1 on lipopolysaccharide (LPS)-induced cardiac dysfunction was examined in a rat model of sepsis. In the absence of CT-1, LPS (1 mg/kg ip) elicited a reduction of systolic function and dilation of the ventricular cavity within 3-6 h after administration. These physiological effects were accompanied by increased ventricular phosphorylation of signal transducers and activators of transcription (STAT) 1 and STAT3, activation of nuclear factor kappaB and expression of iNOS mRNA. Notably, administration of CT-1 (20 microg/kg iv) immediately prior to LPS significantly inhibited all of these LPS-induced changes. To determine whether SOCS1 expression in cardiomyocytes is sufficient to inhibit LPS- and cytokine-induced expression of iNOS mRNA, the effects of forced expression of SOCS1 in cultured neonatal cardiomyocytes were investigated using an adenovirus-mediated transfection system. Forced expression of SOCS1 significantly inhibited iNOS transcription induced by LPS, tumor necrosis factor alpha or interferon-gamma. CONCLUSIONS: CT-1-mediated expression of SOCS1 in cardiomyocytes may be a useful target for preventing sepsis-induced myocardial depression. PMID- 16247221 TI - Down-regulation of endogenous hydrogen sulfide pathway in pulmonary hypertension and pulmonary vascular structural remodeling induced by high pulmonary blood flow in rats. AB - BACKGROUND: The mechanisms responsible for the development of pulmonary hypertension (PH) and pulmonary vascular structural remodeling induced by high pulmonary blood flow are not fully understood. The present study was designed to explore the possible changes in endogenous hydrogen sulfide (H2S), a novel gasotransmitter, on the pathogenesis of PH and pulmonary vascular structural remodeling induced by high pulmonary blood flow. METHODS AND RESULTS: Twenty-two male Sprague-Dawley rats were randomly divided into a shunting group (n=11) and control group (n=11). Rats in the shunting group underwent an abdominal aorta inferior cava vein shunting operation. After 11 weeks of shunting, the plasma level of H2S and lung tissue H2S producing rate were much lower than those of the control group (p<0.01). In situ hybridization analysis showed that the expression of cystathionine gamma-lyase (CSE) mRNA was down-regulated in the pulmonary arteries of the shunting rats compared with the control group (p<0.01), and competitive quantitative reverse transcription-polymerase chain reaction showed that the relative amount of CSEmRNA in lung tissue was decreased significantly (p<0.01). CONCLUSIONS: The endogenous H2S pathway is down-regulated in PH and pulmonary vascular structural remodeling is induced by high pulmonary blood flow. PMID- 16247223 TI - Brain white matter changes during treatment of a child for acute lymphoblastic leukemia. AB - A 13-year old boy with acute lymphoblastic leukemia had bilateral paresis of the upper extremities and aphasia 1 week after high dose methotrexate and triple intrathecal therapy (methotrexate, cytarabin, hydrocortisone). The stroke-like neurological symptoms disappeared on the third day. T2-weighted magnetic resonance imaging showed hyperintensities of white matter on the second day. Despite resolution of the neurological symptoms, magnetic resonance images were still abnormal 3 years after the attack. Methotrexate has been considered to be responsible for ischemic damage to oligodendroglial cells, resulting in demyelination. The changes are occasionally prolonged without persistent neurologic symptoms. PMID- 16247222 TI - Primary aldosteronism combined with preclinical Cushing's syndrome in an elderly patient. AB - Pre-clinical Cushing's syndrome (pre-CS) is sometimes seen with adrenal cortical tumors. An 80-year-old woman had severe hypertension and hypokalemia, the typical clinical features of primary aldosteronism, but detailed hormonal examinations revealed the accompanying pre-CS. After adrenalectomy by laparoscopy, her blood pressure was remarkably reduced and the hypokalemia also recovered. The tumor consisted of mostly light clear cells and scattered dark compact cells resembling islands. Abundant expression of cytochrome P450 aldosterone synthase in the clear cells and cytochrome P450 11beta-hydroxylase in the dark cells was detected by immunohistochemical studies, which confirmed that clear cells can produce aldosterone and compact cells can produce cortisol. PMID- 16247224 TI - Cardiovascular manifestations of Fabry disease and the novel therapeutic strategies. AB - Fabry disease is an inherited lysosomal storage disorder characterized by a pathological intracellular glycosphingolipid deposition. The disease is caused by a deficit in the lysosomal enzyme alpha-galatosidase A, the gene for which is located in the X chrosomal region Xq 22. Globotriaosylceramide (Gb3) accumulate progressively in multi-organ vulnerable cells throughout the body, including cardiovascular, renal, and cerebrovascular systems. The present manuscript is to review cardiovascular and renal manifestations of Fabry disease and the new diagnostic procedures for earlier detection and the therapeutic assessments of this disease. We are applying noninvasive cardiovascular and microcirculation analysis methods and novel cardiac biomarkers. Novel therapeutic strategies for this disease have been developing in recent years, which include the clinically introduced enzyme infusion replacement therapy and experimentally developing gene transfer therapy. We have reported that AAV-mediated muscule-directed gene transfer is very effective for long-term systemic delivery of alpha-gal A (25% of normal mice enzyme activity), resulting in complete clearance of multi-organs Gb3 accumulation. Echocardiographic and immunohistochemical examination demonstrated structural improvement of cardiac hypertrophy. When and to whom the novel therapeutic strategies should be applied to obtain the maximum efficacy and safety remain to be established. PMID- 16247225 TI - Random systematic sextant biopsy versus power doppler ultrasound-guided target biopsy in the diagnosis of prostate cancer: positive rate and clinicopathological features. AB - PURPOSE: To determine the efficacy of power Doppler ultrasound (PDU)in the diagnosis of prostate cancer, the rate of detection of cancer with PDU-guided target biopsy and sextant biopsy, the clinicopathological features of cancer positive specimens, and the relation between these two findings were studied. METHODS: From January 1998 through March 2000, 302 men suspected to have prostate cancer underwent sextant biopsy in association with additional PDU-guided target biopsy. Cases with positive biopsy results were divided into 9 groups as follows: T0: sextant biopsy was positive, but target biopsy was negative; S0: all sextant biopsies were negative, but target biopsy was positive; S1 approximately S6: both sextant biopsy and target biopsy were positive (number indicates number of positive sextant biopsy); Tx: sextant biopsy was positive, but no target biopsy was performed owing to a lack of echogenic abnormalities. The Gleason score (GS) and percent organ confined disease (%OCD) were compared between these 9 groups. RESULTS: Cancer was pathologically detected in 143 of 302 patients (47.4%). PDU detected 39 of 49 digital rectal examination-negative cancers (79.6%) and 5 of 13 transrectal ultrasound-negative isoechoic cancers (38.5%). Of 143 biopsy-positive cases, 6 were in the T0 group (4.2%), 10 in S0 (7.0%), 119 in S1 approximately S6 (83.2%), and 8 in Tx (5.6%). Target biopsy missed 14 (sum of T0 and Tx) cancers, and sextant biopsy missed 10 (S0). The average GS in the Tx group was significantly lower than that in the other groups; consequently, the %OCD was significantly higher. Retrospective analysis revealed that the failure to obtain cancer tissue in 4 of the 6 cases in the T0 group is most likely due to technical failure in obtaining specimens. The overall sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of PDU were 90.2%, 77.4%, 78.2%, 89.8% and 83.4%, respectively. CONCLUSION: PDU in association with sextant biopsy is a useful tool for increasing the rate of detection of prostate cancer. Further advances in ultrasound technology may enable the detection of prostate cancer by target biopsy alone and consequently may reduce the number of unnecessary biopsies. However, PDU-guided target biopsy alone is insufficient for cancer detection at the present time because of possible technical failure in obtaining specimens and the existence of PDU-negative cancer. Although more evidence is required, PDU-negative cancer is suggested to be less aggressive clinically, possibly justifying a watch and wait policy. PMID- 16247226 TI - Hypermethylation of the putative tumor-suppressor genes DCC, p51/63 and O6 methylguanine-DNA methyltransferase (MGMT) and loss of their expressions in cell lines of hematological malignancies. AB - Alterations and defective expression of three putative tumor-suppressor genes, the deleted in colorectal cancer (DCC), p51, and O(6)-methylguanine-DNA methyltransferase (MGMT), have been demonstrated in many cancers. However, it is not known whether the defective expression of each of these genes is independent or whether it reflects a specific methylation abnormality. Here, we investigated the expression of the DCC, p51 and MGMT genes and the methylation status of the 5' flanking CpG region in 17 cell lines established from hematological malignancies. The reverse transcriptase-polymerase chain reaction method showed DCC expression to be absent in 13 of the 17 cell lines and showed expression of both p51 and MGMT to be absent in 5 of the 17 cell lines. The methylation patterns were analyzed with methylation-specific polymerase chain reaction (MSP) of the 5'flanking region of the DCC and p51 genes and the promoter region of the MGMT gene. Although unmethylated patterns of the CpG region in the DCC, p51, and MGMT genes were observed in all 11 normal controls, abnormal methylation patterns of these genes were found even in many cell lines expressing these genes. A hypermethylation pattern was detected for the CpG region of MGMT and p51 in cells that did not express these genes. In contrast, a hypermethylation pattern was not always detected for the CpG region of DCC in cells with reduced DCC expression. The results of this study indicate that in many hematological cell lines, the DCC, p51, and MGMT genes have been abnormally methylated in the CpG region. Hypermethylation of these three genes may be independent events in each cell line. PMID- 16247227 TI - The role of transanal endoscopic microsurgery for rectal tumors. AB - PURPOSE: The management of rectal tumors is complex, because of the balance between preserving rectoanal function and curing the patient. Transanal endoscopic microsurgery (TEM) is both an effective treatment for benign rectal tumors and early cancers, and a diagnostic tool for determining tumor depth, or for residual tumors of post endoscopic mucosal resection. In the present study, we evaluated the role of TEM in the management of rectal tumors. METHODS: Twenty six patients with rectal tumors underwent TEM from December 2000 through March 2005 in our department. The operations were performed by a single surgeon, and the indications were mainly limited to a) benign tumors for which endoscopic resection was difficult, b) early cancers that had invaded the submucosa within 500 microm of the muscularis mucosae, c) submucosal tumors, i.e., gastrointestinal stromal tumor, carcinoid tumors, d) local excision for diagnosis, and e) palliative resection for high-risk cases. Anesthesia, operation time, sizes of the tumor and of resected specimens, postoperative complications, length of hospitalization, pathological results, and postoperative recurrence rate were reviewed. RESULTS: The mean age of patients was 61.9 years, and the cases included 14 rectal cancers, 7 adenomas, 1 gastrointestinal stromal tumor, and 3 rectal carcinoid tumors. The mean operation time was 96 min (range, 40 approximately 235 min.). The average postoperative hospital stay was 4.8 days. All tumors were resected with horizontal and vertical safety margin. The mean size of the resected specimens was 9.0 cm(2). In one case, the tumor had infiltrated the proper muscle layer, as shown by intraoperative frozen sectioning, which necessitated abdominoperineal resection. In 3 cases, pathological examination revealed massive infiltration into the submucosal layer. 2 patients underwent low anterior resection, and the remaining patient refused additional surgery despite our recommendation. No deaths occurred. No major postoperative complications were noted. The mean follow-up period was 27.2 months. Only one case of lymph node metastasis was observed, in the left iliac lymph node 3 years after TEM. CONCLUSIONS: TEM is a minimally invasive surgical procedure for rectal tumors, which allows the whole depth of the rectal wall to be resected with a safety surgical margin. Although TEM requires technical skill and accurate preoperative diagnosis, the procedure is safe, facilitates accurate diagnosis of tumor depth, and limits the need for additional surgery. PMID- 16247228 TI - Sudden death in a case of cardiac amyloidosis immediately after pacemaker implantation for complete atrioventricular block. AB - We report a patient with cardiac involvement associated with primary amyloidosis presenting marked left ventricular (LV) wall thickening, severely decreased systolic and diastolic function, and complete atrioventricular block (CAVB), who died suddenly of cardiac arrest caused by electro-mechanical uncoupling occurring immediately after permanent pacemaker implantation. Post mortem examination showed no procedural complications such as cardiac or venous perforation. The heart was densely infiltrated with amyloid fibrils, especially in the extracellular tissues surrounding the papillary vessels. PMID- 16247229 TI - A case report of rapidly progressing cauda equina symptoms due to rheumatoid arthritis. AB - Although rheumatoid involvement of the lumbar spine is relatively rare, we report a patient with rapidly progressing cauda equina symptoms due to rheumatoid diskitis. A 72-year-old woman was admitted to our hospital because of motor weakness below the iliopsoas muscle and sensory disturbance beneath the level of L2. Plain X-ray films, computed tomography, and magnetic resonance imaging demonstrated destruction of the L2/3 intervertebral disc and endplates with subluxation of the facet joints. The dural sac was compressed. Based on a diagnosis of spinal canal stenosis due to rheumatoid diskitis, we performed partial laminectomy and posterolateral fusion with pedicle screws. The neurological deficits improved immediately. The mechanism of intervertebral disc destruction in this case is thought to be rheumatoid nodes or enthesitis. Destruction of the facet joints and the intervertebral disc might have led to severe instability and spinal canal stenosis. PMID- 16247230 TI - A case of cervical spondylitis during stellate ganglion block. AB - We report a case of cervical spondylitis that developed during treatment with a series of stellate ganglion blocks. A 65-year-old man was scheduled for 10 sessions of stellate ganglion block for treatment of right-sided deafness of sudden onset due to Ramsay Hunt syndrome. Administration of betamethasone was started 5 days before the first block and was continued for 6 weeks. After disinfection of the skin by povidone iodine, each stellate ganglion block was performed via the paratracheal approach. The first four block sessions were uneventful. However, during the fifth session, the patient complained of neck pain. After 10 sessions, the deafness improved and the patient was discharged from the hospital. Three weeks after discharge, he was readmitted for sustained neck and bilateral shoulder pain and numbness of the right hand. Cervical roentgenography and magnetic resonance imaging revealed spondylitis of C5 and C6. Antibiotics were administered for 2 weeks. The inflammatory variables on blood examination improved, but cervical roentgenography performed 8 weeks after the last block showed that the vertebral body of C6 was nearly completely destroyed. Four months after the last block, the vertebral bodies of C5 and C6 had fused. This case indicates that when stellate ganglion block is performed in patients who are taking a corticosteroid, the disinfection procedure must be strictly followed and that if the patient complains of neck or shoulder pain, cervical roentgenography or magnetic resonance imaging or both should be immediately performed to assess the presence of spondylitis. PMID- 16247231 TI - Blood concentration of propofol in a patient with delayed emergence from propofol nitrous oxide anesthesia. AB - We measured the blood concentration of propofol in a patient with delayed emergence from propofol-nitrous oxide anesthesia. A 78-year-old man underwent subtotal gastrectomy under both epidural and propofol-nitrous oxide anesthesia and did not regain consciousness soon after termination of propofol infusion. Preoperative laboratory examination revealed anemia and a low blood total protein concentration, but there was no evidence of impaired liver function. While the anesthesiologists were waiting for the patient to regain consciousness, a surgeon mentioned that the common hepatic artery might have been occluded during surgical manipulation. Arterial blood samples were obtained 50 and 80 minutes after termination of propofol infusion, and the blood concentration of propofol was measured. We considered that clearance of propofol through the hepatic route may have been impaired; however, the actual blood concentrations of propofol were not significantly increased compared with the respective blood concentrations obtained in the simulation. Therefore, the acute liver damage did not significantly impair elimination of propofol. Because most propofol molecules in the blood bind to proteins and erythrocytes, it is suspected that the anemia and low blood total protein concentration led to an increase in the free fraction of propofol in the blood, thereby delaying emergence from anesthesia. PMID- 16247232 TI - Spurt bleeding from a calcificated gastrointestinal stromal tumor in the stomach. AB - Calcifications within primary gastrointestinal tumors are rare. Gastrointestinal stromal tumor (GIST) is an unusual nonepithelial tumor that develops in the gastrointestinal tract. In this paper we describe a case of spurt bleeding from a calcificated GIST in the stomach successfully treated by partial gastric resection. A 77-year-old man was admitted for chest discomfort and loss of consciousness. Endoscopic examination revealed spurt bleeding from the top of the submucosal tumor. No other lesions or points of bleeding were found in the stomach. Emergency partial gastrectomy was performed, and the stomach was closed. The cut surface of the tumor had a firm, solid, whitish-gray parenchyma with patchy calcification. Microscopic observation revealed a profusion of spindle shaped tumor cells with calcification growing from the gastric muscular propria to the submucosa. The cells exhibited low mitotic activity and no prominent signs of nuclear atypia. Immunohistochemical staining of the tumor demonstrated positive reactivity for CD34, KIT, and vimentin, but negative reactivity for alpha-smooth muscle actin, desmin, and S-100 protein. Tumor cells positive for Mib-1 were rare. The diagnosis of the tumor was established as GIST. PMID- 16247233 TI - A rapidly enlarging nocardial brain abscess mimicking malignant glioma. AB - Nocardial brain abscesses are uncommon and are not preceded by clear infectious symptoms in most cases. Delayed identification of the bacteria is responsible for a high mortality rate. A 58-year-old afebrile woman was admitted to our hospital because of progressive right hemiparesis and aphasia. Magnetic resonance imaging (MRI) showed a single ring-enhanced lesion in the left frontal lobe. It was extremely difficult to establish the diagnosis of brain abscess, because the laboratory data provided little evidence of bacterial infection, (201)TlCl scintigraphy revealed definite accumulation of thallium in the lesion, and follow up MRI demonstrated rapid enlargement of the lesion. Total resection was performed because of the possibility of a malignant brain tumor, but brain abscess was finally diagnosed with histological examination. A nocardial species was detected through microscopic examination of the pus obtained at surgery, and this precise diagnosis of nocardial brain abscess in the early stage enabled the administration of appropriate antibiotics and the patient's quick recovery. Nocardial brain abscesses are often misdiagnosed as malignant brain tumors, and a definitive diagnosis may not be possible without detecting bacteria from the lesion. Total excision of the abscess can produce good results when the abscess is large and located superficially, but incomplete aspiration and drainage of a lesion is associated with a high chance of relapse. PMID- 16247234 TI - Prevalence of asymptomatic microbleeds in patients with moyamoya disease. AB - Basal moyamoya vessels are a potential source of hemorrhage in patients with moyamoya disease, but the etiology remains unclear. Symptomatic hemorrhage resulting from long-standing hemodynamic effects on pathologically dilated, fragile moyamoya vessels may be preceded by asymptomatic microbleeding in adult moyamoya disease patients, regardless of hemorrhagic or ischemic onset. T2* weighted magnetic resonance (MR) imaging was used to investigate the presence of microbleeds in 27 adult patients with angiographically confirmed moyamoya disease, 21 females and six males aged 18-70 years (mean 40.8 +/- 15.7 years). Clinical diagnosis was intracranial bleeding in six patients, transient ischemic attack or cerebral infarction in 18, and asymptomatic in three. Asymptomatic microbleeds were detected in four of the 27 patients, two of six who initially presented with hemorrhagic events and two of 18 with ischemic onset. These microbleeds were located in the paraventricular white matter, temporal subcortex, and basal ganglia. The presence of microbleeds had no correlation with either patient age or duration from disease onset or diagnosis of disease. A large cohort study is needed to explore the significance of asymptomatic microbleeds in moyamoya disease. PMID- 16247236 TI - Craniovertebral junction realignment for the treatment of basilar invagination with syringomyelia: preliminary report of 12 cases. AB - Twelve selected patients, eight males and four females aged 14 to 50 years, with syringomyelia associated with congenital craniovertebral bony anomalies including basilar invagination and fixed atlantoaxial dislocation, and associated Chiari I malformation in eight, were treated by atlantoaxial joint manipulation and restoration of the craniovertebral region alignment between October 2002 and March 2004. Three patients had a history of trauma prior to the onset of symptoms. Spastic quadriparesis and ataxia were the most prominent symptoms. The mean duration of symptoms was 11 months. The atlantoaxial dislocation and basilar invagination were reduced by manual distraction of the facets of the atlas and axis, stabilization by placement of bone graft and metal spacers within the joint, and direct atlantoaxial fixation using an inter-articular plate and screw method technique. Following surgery all patients showed symptomatic improvement and restoration of craniovertebral alignment during follow up from 3 to 20 months (mean 7 months). Radiological improvement of the syrinx could not be evaluated as stainless steel metal plates, screws, and spacers were used for fixation. Manipulation of the atlantoaxial joints and restoring the anatomical craniovertebral alignments in selected cases of syringomyelia leads to remarkable and sustained clinical recovery, and is probably the optimum surgical treatment. PMID- 16247235 TI - Awake surgery for glioma resection in eloquent areas--Zurich's experience and review--. AB - Awake surgery was performed in a series of 21 patients with gliomas in eloquent areas with the use of intraoperative electrical mapping. Gross total removal was performed in 18 patients. There was no operative mortality. Postoperative findings included no change in symptoms and signs in 10 patients, improvement of the preoperative deficit in 11 patients. Four patients had improved Karnofsky performance status (KPS) scores after surgery, 17 patients were stable, and no patient had lower KPS score. Extensive radical resection of gliomas prolongs the overall survival and improves the patient's quality of life. However, surgical resection of gliomas located within the sensorimotor or language areas remains a neurosurgical challenge in reducing eloquent neurological sequelae. Awake surgery with intraoperative functional mapping is a safe approach to maximize the extent of tumor removal and to minimize the resultant neurological deficits in the treatment of glioma involving the eloquent cortex. PMID- 16247237 TI - Subdural hematoma associated with dural metastasis--case report--. AB - A 56-year-old female presented with acute subdural hematoma associated with dural metastasis. The patient had been treated for breast cancer with disseminated bone and lung metastases. Evacuation of the hematoma with local management of the tumor and bleeding successfully improved her neurological condition and she underwent postoperative radiotherapy. This condition is especially associated with dural metastasis from adenocarcinoma (most frequently stomach cancer) and the clinical outcome depends on the general condition of the patient and the status of the coagulation disorders. If the tumors are multiple, as in this case, extreme caution should be paid to recurrent bleeding in the ipsilateral or contralateral side. PMID- 16247238 TI - Multiple aneurysms arising from the posterior inferior cerebellar artery with unknown etiology--case report--. AB - A 67-year-old man presented with multiple aneurysms arising from the caudal loop of the posterior inferior cerebellar artery (PICA), possibly as a result of blunt trauma. Computed tomography of the head revealed subarachnoid hemorrhage in the posterior fossa and sylvian fissure. Repeated angiography demonstrated an aneurysmal dilatation and an irregular wall on the caudal loop of the PICA. Under the operating microscope, two lesions were observed 10 mm distal to the apex of the caudal loop, both consisting of a tiny hole on the vessel wall with a fragile fringe of connective tissue and covered with a firm clot. The height of the lesions corresponded to the C-l lamina, so the lesions were probably traumatic rather than saccular. PMID- 16247239 TI - Pseudocapsule formation after gamma knife radiosurgery for trigeminal neurinoma- case report--. AB - A 38-year-old female presented with a trigeminal neurinoma manifesting as left facial paresthesia. The diagnosis was based on magnetic resonance (MR) imaging findings. Gamma knife radiosurgery (GKR) was performed at another hospital at her request. Fifteen months after the GKR, follow-up MR imaging revealed tumor regrowth causing extensive compression of the brainstem, and cyst formation in the tumor. Her clinical symptoms including facial pain and diplopia had worsened, so she was referred to our affiliated hospital for microsurgery. The tumor was totally resected, but the left trigeminal nerve had to be sacrificed because of pseudocapsule formation which covered both the tumor and the trigeminal nerve fibers. The diplopia disappeared, but her facial pain deteriorated after the operation. GKR can induce fibrosis or degenerative change in nearby structures, which may complicate subsequent surgery. PMID- 16247240 TI - Supratentorial primitive neuroectodermal tumor in an aged patient--case report--. AB - An 88-year-old woman presented with a supratentorial primitive neuroectodermal tumor (PNET) manifesting as disturbance of consciousness and left hemiplegia. Magnetic resonance imaging showed a large mass lesion in the right frontotemporal region. She underwent biopsy of the lesion that confirmed the diagnosis of PNET. Her poor condition only allowed chemotherapy with methyl 6-[3-(2-chloroethyl)-3 nitrosoureido]-6-deoxy-alpha-D-glucopyranoside (MCNU), vincristine, and prednisolone to be performed. The patient died approximately 6 months after diagnosis due to enlargement of the tumor. Supratentorial PNET is a rare tumor, especially in adults. Multimodal therapy consisting of gross total or subtotal resection, radiation therapy, and chemotherapy is generally considered necessary for patients with supratentorial PNET. However, the condition of each patient should be considered in determining the therapeutic plan, especially in the case of extremely aged patients, since supratentorial PNET is malignant and long-term survival is rare despite aggressive treatment. PMID- 16247241 TI - Bilateral vocal cord paralysis in newborns with neuraxial malformations--two case reports--. AB - Two neonates presented with inspiratory stridor due to bilateral vocal cord paralysis associated with occipital encephalocele, Chiari malformation, and hydrocephalus in one patient, and cervical meningomyelocele and Chiari malformation in the other patient. The clinical symptoms dramatically regressed after repair of the encephalocele or meningomyelocele with no requirement for craniovertebral decompressive procedures or shunts in the acute phase. Careful evaluation of neonatal stridor and recognition of vocal cord paralysis are important, as treatment of associated congenital central nervous system anomalies is likely to achieve satisfactory surgical results. PMID- 16247242 TI - Spontaneous cervical paraspinal and epidural giant abscess in a child--case report--. AB - A 10-year-old girl presented with a very rare paraspinal and spinal epidural abscess manifesting as fever, and neck pain and stiffness. Initially, she was treated under a diagnosis of meningitis for 3 weeks. However, she developed monoparesis of the right upper extremity, and was referred for neurosurgery. Magnetic resonance imaging revealed an epidural and paraspinal lesion intensely enhanced by gadolinium. The patient underwent urgent surgery for C2-3 laminectomy and abscess drainage, followed by broad spectrum antibiotic therapy. She was discharged and followed up in the outpatient clinic. Two months later, the paraspinal abscess recurred with great increase in size. A second operation was performed and 150 ml pus was drained. Streptococcus anginosus was grown in the culture. The patient fully recovered after long-term targeted antibiotic therapy. Such abscesses are very rare in children, especially in the cervical region. The correct diagnosis can be difficult to establish but early treatment is essential for a good prognosis. PMID- 16247243 TI - Peritoneal shunt tube migration into the stomach--case report--. AB - A 47-year-old man presented with repeated headache and feverishness 3.5 years after undergoing ventriculoperitoneal shunt surgery for normal pressure hydrocephalus secondary to subarachnoid hemorrhage. Abdominal computed tomography revealed that the peritoneal catheter was encased by fibrous tissue and the distal end of the catheter had migrated into the stomach. The diagnosis was spontaneous gastric perforation by the ventriculoperitoneal shunt. The fibrous tissue was expected to seal the very small gastric perforation, so the catheter was successfully extracted through a scalp incision without abdominal surgical intervention. PMID- 16247244 TI - Novel drug delivery systems: potential in improving topical delivery of antiacne agents. AB - Acne is the most common cutaneous disorder of multifactorial origin with a prevalence of 70-85% in adolescents. The majority of the acne sufferers exhibit mild to moderate acne initially, which progresses to the severe form in certain cases. Topical therapy is employed as first-line treatment in mild acne, whereas for moderate and severe acne, systemic therapy is required in addition to topical therapy. Currently, several topical agents are available that affect at least one of the main pathogenetic factors responsible for the development of acne. Although topical therapy has an important position in acne treatment, side effects associated with various topical antiacne agents and the undesirable physicochemical characteristics of certain important agents like tretinoin and benzoyl peroxide affect their utility and patient compliance. Novel drug delivery strategies can play a pivotal role in improving the topical delivery of antiacne agents by enhancing their dermal localization with a concomitant reduction in their side effects. The current review emphasizes the potential of various novel drug delivery strategies like liposomes, niosomes, aspasomes, microsponges, microemulsions, hydrogels and solid lipid nanoparticles in optimizing and enhancing the topical delivery of antiacne agents. PMID- 16247245 TI - The role of menthol in skin penetration from topical formulations of ibuprofen 5% in vivo. AB - In vivo plasma profiles from formulations containing 5% ibuprofen were compared after a single topical application in a randomised, double-blind, cross-over trial. Ibuleve gel (Dermal Laboratories, UK) contained only ibuprofen whilst Deep Relief gel (Mentholatum, UK) also contained 3% menthol. In contrast to results obtained when these products were compared under in vitro conditions, there was no statistically significant difference in vivo between delivery of ibuprofen. Estimated relative bioavailability fraction (Deep Relief gel/Ibuleve gel) from log-transformed AUC((0-24h)) was 0.99 (95% CI: 0.94-1.04), estimated C(max )ratio was 0.96 (95% CI: 0.91-1.00) and estimated t(max) ratio was 1.01 (95% CI: 0.81 1.20). Menthol produces local vasodilation, which reduces skin barrier function, and these data demonstrate that it is inappropriate to extrapolate from in vitro data where formulation components produce biologically-mediated enhancement of permeation which cannot be modelled ex vivo. In clinical use, these products deliver comparable amounts of ibuprofen, but only Deep Reliefgel provides the secondary immediate benefit of the direct analgesic action of menthol. PMID- 16247246 TI - Evaluation of skin viscoelasticity in type 1 neurofibromatosis patients. AB - Neurofibromatosis type 1 (NF1) is a frequent autosomal dominant disease characterized by cutaneous benign tumors called neurofibromas. Surgery takes an important place in managing these skin disorders. However, skin distensibility and softness of NF1 patients quickly offset the surgical benefit. The aim of this study was to determine the rheological behavior of neurofibromas and compare it with healthy skin in an attempt to comprehend what leads to this phenomenon. Thirty patients were admitted to this study. A group of 24 healthy control subjects was also included. The skin elasticity was assessed by a noninvasive in vivo suction device (Cutometer) including 5 consecutive suctions. The assessments were performed on neurofibroma skin, the supposedly healthy skin around neurofibromas and the healthy skin of control subjects. The extensibility at the first and the fifth traction in NF1 patients (neurofibromas and the supposedly healthy skin around it) was significantly different compared to the healthy skin of control subjects. The viscoelastic parameters obtained from the neurofibromas were significantly different in comparison to those obtained from the supposedly healthy skin of NF1 patients and the healthy skin of control subjects. The rheological profiles of the neurofibromas and the apparent healthy skin of NF1 patients demonstrated a hyperextensibility behavior, but in neurofibromas, the skin was unable to return to its initial position at the end of the stretch. PMID- 16247247 TI - Transdermal delivery of tea catechins by electrically assisted methods. AB - Tea polyphenols, including (+)-catechin, (-)-epicatechin, and (-) epigallocatechin-3-gallate (EGCG), have been shown to possess potent antioxidant and chemopreventive activities. The aim of this study was to assess the effects of electroporation, iontophoresis, and their combination on the transdermal delivery of tea catechins across porcine skin. The permeation characteristics were investigated using various analogues of catechins, pH values, and modes of electroporation and iontophoresis. The mechanisms by which these catechins were transported via the skin were elucidated by examining the electric conductivity, transepidermal water loss (TEWL), and fusion of stratum corneum lipid liposomes (SCLL). The isomers, (+)-catechin and (-)-epicatechin, showed different behaviors of skin permeation and local skin deposition with the electrically assisted methods. The results suggest evidence of selective skin absorption of (-) epicatechin over (+)-catechin. A synergistic effect was detected for (+)-catechin but not for (-)-epicatechin after application of electroporation followed by iontophoresis. The presence of a gallic acid ester in the structure of EGCG significantly increased the skin uptake of catechins. However, a negligible amount of or no EGCG molecules permeated across the skin. The mechanisms involved in the enhancement of electroporation may be the skin reservoir effect and an increase in skin permeability. The TEWL profiles suggest that in addition to the force of electrorepulsion, the skin hydration effect and structural alterations may also have contributed to the enhancement by iontophoresis. Electroporation did not influence the skin barrier function, although the skin permeability increased according to the SCLL fusion study. PMID- 16247248 TI - Are sweat glands an alternate penetration pathway? Understanding the morphological complexity of the axillary sweat gland apparatus. AB - To build an effective barrier against the penetration of extrinsic agents is one of the skin's main functions. The barrier properties of the stratum corneum and the epidermis have been subject to extensive studies in the past while the role of skin appendages as possible pathways of penetration are only rarely described. In order to study the possible penetration barriers in these complex appendages, a careful investigation of their morphology and ultrastructure has to be done. Studying the morphology of axillary skin appendages requires clear-cut criteria for the differentiation between eccrine, apocrine and apoeccrine glands. Therefore we studied the distribution of proteins described to be specific for either eccrine or apocrine glands (CD15, CD44, S-100 and milk fat globulin) on axillary skin samples from healthy young adults by immunofluorescence. Additionally, we examined the distribution of cytoskeletal proteins such as cytokeratins (1/10/11, 14, 18) and F-actin. For a more detailed understanding of the possible versatile barrier elements of the axillary sweat glands, we studied the distribution of tight-junction-associated proteins (occludin, claudin 1, claudin 4). The coils and the dermal duct may provide an active barrier built of tight junctions as occludin and claudin 4 are co-localized. However, the intra epidermal duct did not show any co-localization of the investigated proteins. By combining morphological features as revealed by F-actin staining and the distribution of the above-mentioned proteins, immunocytochemical typing of eccrine and apocrine glands becomes possible. With this tool, we could also confirm the existence of apoeccrine glands and locate them in their 'natural environment'. PMID- 16247249 TI - Activity of the triazole antifungal r126638 as assessed by corneofungimetry. AB - BACKGROUND: R126638 is a novel triazole exhibiting potent in vitro and in vivo antifungal activity against fungal pathogens including dermatophytes and yeasts. OBJECTIVE: To determine the antifungal activity in time in the stratum corneum of healthy volunteers after oral intake of R126638 at a daily dose of 100 or 200 mg for 1 week. METHOD: Sixteen male volunteers were randomly allocated to oral treatment with either 100 or 200 mg of R126638 once daily for 1 week. Five cyanoacrylate skin surface strippings (CSSS) were obtained from the forearm of each subject before drug intake at day 1. CSSS were also collected during treatment at day 2 (24 h after the first drug intake, before the second drug intake), at day 4 (before the fourth drug intake) and at day 7 (10 h after the last drug intake). The post-treatment lingering effect was assessed at day 10 (3 days after treatment) and at day 14 (7 days after treatment). The corneofungimetry bioassay was performed on these CSSS to assess the antifungal profile of R126638. Cells of different fungal species (Trichophyton rubrum, Trichophyton mentagrophytes, Microsporum canis, Candida albicans and Malassezia globosa) were deposited and cultured for 10 days on CSSS in a sterile and controlled environment. The extent of fungal growth on the stratum corneum was determined using computerized image analysis. RESULTS: R126638 clearly reduced the growth of all tested fungal species. The onset of effects of R126638 was evidenced at day 4 when it reached statistical significance for 3 of 5 species. At day 7, significance was reached for 4 of 5 species. During the posttreatment period, R126638 remained effective for 4 of 5 species at day 10, and this activity persisted until day 14 for 2 of 5 species. CONCLUSION: A broad spectrum antifungal activity was rapidly expressed in the stratum corneum after oral intake of R126638. The drug likely reached the upper layers of the stratum corneum by diffusion and persisted in this location for at least 7 days after treatment. PMID- 16247250 TI - Epidemiology of children's individual sports injuries. An important area of medicine and sport science research. PMID- 16247251 TI - Equestrian injuries. AB - OBJECTIVE: This chapter reviews the current evidence for the epidemiology of pediatric equestrian injuries. DATA SOURCES: The relevant literature was searched through the use of MEDLINE (1966-2004) and SPORT DISCUS (1975-2004) searches, hand searches of journals and reference lists and discussions with experts and sporting organizations worldwide. Keywords and Mesh headings used in all searches included horse racing, children, pediatric injuries, sports injuries, equestrian injuries and sports trauma. MAIN RESULTS: Limited data exist on the epidemiology of pediatric equestrian injuries. Most studies note the high preponderance of females with a peak incidence at approximately 14 years of age. This is likely to reflect the higher rate of female riders. The two most common horse riding related injuries are long bone fractures and head injury. Although most injuries occur during recreational riding, approximately 15% of injuries occur in nonriding activities such as feeding, handling, shoeing and saddling. CONCLUSIONS: While there is little knowledge of injury demographics or the efficacy of prevention countermeasures in this field, it is likely that injuries will continue to occur. The major challenge in reducing pediatric equestrian injuries is the formal scientific demonstration that the various proposed injury prevention measures are effective. With the majority of equestrian injuries happening during unsupervised leisure riding, the prospect of injury prevention is limited. PMID- 16247252 TI - Gymnastics injuries. AB - OBJECTIVE: The purpose of this chapter is to review the distribution and determinants of injury rates as reported in the pediatric gymnastics injury literature, and to suggest measures for the prevention of injury and directions for further research. DATA SOURCES: An extensive search of Pubmed was conducted using the Text and MeSH words "gymnastics" and "injury" and limited to the pediatric population (0-18 years). The review focused on studies using denominator-based designs and on those published in the English language. Additional references were obtained from hand searches of the reference lists. Unpublished injury data from the USA Gymnastics National Women's Artistic Gymnastics Championships during 2002-04 were also analyzed. MAIN RESULTS: Comparison of study results was compromised due to the diversity of study populations, variability of injury definition across studies, and changes in rules and equipment across years. Notwithstanding, this review of the literature reveals a reasonably consistent picture of pediatric gymnastics injuries. The incidence and severity of injuries is relatively high, particularly among advanced level female gymnasts. Body parts particularly affected by injury vary by gender and include the ankle, knee, wrist, elbow, lower back, and shoulder. Ankle sprains are a particular concern. Overuse and nonspecific pain conditions, particularly the wrist and low back, occur frequently among advanced-level female gymnasts. Factors associated with an increased injury risk among female gymnasts include greater body size and body fat, periods of rapid growth, and increased life stress. CONCLUSIONS: Above all, this overview of the gymnastics injury literature underscores the need to establish large-scale injury surveillance systems designed to provide current and reliable data on injury trends in both boys and girls gymnastics, and to be used as a basis for analyzing injury risk factors and identifying dependable injury preventive measures. PMID- 16247253 TI - Martial arts injuries. AB - OBJECTIVE: To review the current evidence for the epidemiology of pediatric injuries in martial arts. DATA SOURCES: The relevant literature was searched using SPORT DISCUS (keywords: martial arts injuries, judo injuries, karate injuries, and taekwondo injuries and ProQuest (keywords: martial arts, taekwondo, karate, and judo), as well as hand searches of the reference lists. MAIN RESULTS: In general, the absolute number of injuries in girls is lower than in boys. However, when expressed relative to exposure, the injury rates of girls are higher. Injuries by body region reflect the specific techniques and rules of the martial art. The upper extremities tend to get injured more often in judo, the head and face in karate and the lower extremities in taekwondo. Activities engaged in at the time of injury included performing a kick or being thrown in judo, while punching in karate, and performing a roundhouse kick in taekwondo. Injury type tends to be martial art specific with sprains reported in judo and taekwondo and epistaxis in karate. Injury risk factors in martial arts include age, body weight and exposure. CONCLUSIONS: Preventive measures should focus on education of coaches, referees, athletes, and tournament directors. Although descriptive research should continue, analytical studies are urgently needed. PMID- 16247254 TI - Skiing and snowboarding injuries. AB - OBJECTIVE: To critically examine the literature on skiing and snowboarding injuries in children and adolescents. DATA SOURCES: Searched English language articles from: Medline, SPORTDiscus, Cumulative Index to Nursing and Allied Health Literature, Current Contents, and HealthSTAR. The table of contents for Ski Trauma and Skiing Safety Series published by the American Society for Testing and Materials were also examined. MeSH headings included: Sports, Athletic Injuries, and Accidents. Keywords used within these headings were Skiing and/or Snowboarding with focus on children, adolescents, youth, students, or age group related comparisons. MAIN RESULTS: The patterns and rates of injury differed markedly by activity and study design. Most studies were case-series investigations providing little useful information on risk factors. Intrinsic risk factors included: lower ability, younger age, past injury, and female sex. Extrinsic risk factors were improper binding adjustment, no helmet, certain slope characteristics, and no wrist guards. The literature on the effect of activity, equipment ownership and lessons on injury risk was equivocal. CONCLUSIONS: Suggestions for injury prevention include the use of helmets and wrist guards, participation on appropriate runs for ability level, proper fit and adjustment of bindings and other equipment, and taking lessons with the goal of increasing ability and learning hill etiquette. Many areas requiring further research are identified and discussed. New methodological approaches hold promise in advancing the field of ski and snowboard injury research. PMID- 16247255 TI - Tennis injuries. AB - OBJECTIVE: The purpose of this chapter is to critically review the existing studies on the epidemiology of tennis injuries in pediatric athletes, present suggestions for the prevention of injury based on these studies, and present suggestions for future research. DATA SOURCES: Data sources included published articles on pediatric tennis injuries, a previously published review by the authors, and unpublished data from one of the authors (MS). MAIN RESULTS: Most studies of tennis injuries show that they are of microtrauma origin, develop over time, and result in short times of absence from play. They involve all joints of the body, but have a higher incidence in the shoulder, back, and knee. Intrinsic and extrinsic risk factors may be related to the incidence of injury. These factors may be evaluated by a comprehensive pre-participation exam, and preventive strategies may be implemented. CONCLUSIONS: Most injury studies in pediatric tennis players vary in the population studied, methods of injury evaluation, and risk factors studied. Consequently, few specific conclusions can be derived about the causative factors. Further longitudinal prospective studies need to be done to completely discover all the factors involved in producing tennis injuries. PMID- 16247256 TI - Track and field injuries. AB - OBJECTIVE: A review of the existing literature on injuries to youth (< or =18 years old) in track and field or athletics. DATA SOURCES: Searches of the Medline and SPORT Discus databases for English language articles through the end of 2003, using the search terms (adolescent or youth) and (track or field or running) and injuries. MAIN RESULTS: Only nine prospective or retrospective studies were found dealing with track and field injuries in children and that stated injury rates or provided enough information to allow the estimation of injury rates. Differences in study design and inconsistencies in the definition of a reportable injury provided major hindrances to making comparisons or combining data across studies. Among the few conclusions that can be drawn are that the lower extremities account for the majority of injuries, and muscle strains and ligament sprains are the predominant types of injury. While a majority of injuries may occur during training, since there is much more exposure during training than during competitions, the risk of injury is about four times higher during competitions. CONCLUSIONS: Informed decisions with regard to preventing injuries in youth track and field are dependent upon the quality of the basic epidemiological data available, and at this time such data are, for the most part, nonexistent. Because of the large numbers of participants and the large number and variety of activities involved in track and field, adequately designed epidemiological research is difficult, but opportunities for research in this sport are available for anyone willing to take on the challenge. PMID- 16247257 TI - Wrestling injuries. AB - OBJECTIVE: The purpose of this chapter is to review critically the existing studies on the epidemiology of pediatric wrestling injuries and to discuss suggestions for injury prevention and further research. DATA SOURCES: Data were obtained from the sports medicine and science literature since 1951. Literature searches were performed using the National Library of Medicine, Pubmed, Medline, Grateful Med, Sports Sciences, SportsDiscus. Keywords used included "Wrestling, Wrestle, Wrestling Injuries, Fractures, and Dermatologic". MAIN RESULTS: Only eight prospective or retrospective studies were found dealing with pediatric wrestling injuries and that provided sufficient information to allow the estimation of injury rates. Exposure-based injury rates were between 6.0 and 7.6 injuries per 1,000 athletic-exposures. Injury rates increased with age, experience, and level of participation. The head/spine/trunk was the body region that incurred the greatest frequency of injuries, followed by the upper and lower extremities. CONCLUSIONS: There are several potential areas for decreasing injury risk in wrestlers, including equipment, coaching, officiating and training. However, informed decisions with regard to preventing injuries are dependent upon the quality of the basic epidemiological data available, and at this time, analyses of risk factors and potential preventive measures are lacking. PMID- 16247258 TI - Injury prevention and future research. AB - OBJECTIVES: To critically examine and summarize the literature identifying risk factors and prevention strategies for injury in child and adolescent sport. DATA SOURCES: Seven electronic databases were searched including: Medline, Cumulative Index to Nursing and Allied Health Literature (CINAHL), Psychinfo, Cochrane Database for Systematic and Complete Reviews, Cochrane Controlled Trials Registry, HealthSTAR and SPORTDiscus. Medical subject headings and text words included: athletic injury, sport injury, risk factors, adolescent and child. Additional articles were reviewed based on sport-specific contributions in the previous chapters of this book. MAIN RESULTS: Despite the diversity of injuries occurring in various pediatric sporting populations, the uniformity with respect to many of the risk factors identified in the literature is noteworthy (i.e. previous injury, age, sport specificity, psychosocial factors, decreased strength and endurance). The literature is significantly limited with respect to the prospective evaluation of risk factors and prevention strategies for injury in pediatric sport. The consistencies, however, between the adult and pediatric literature are encouraging with respect to prevention strategies involving neuromuscular training programs (i.e. balance training programs) to reduce lower extremity injuries in some sports and the use of sport-specific protective equipment (i.e. helmets). CONCLUSIONS: Notwithstanding the limitations in the literature, the successful evaluation of some sport-specific prevention strategies to reduce injury in pediatric sport is encouraging. There is significant opportunity to methodologically improve upon the current pediatric sport injury literature in descriptive surveillance research, risk factor evaluation research, and prevention research. There is a need for prospective studies, ideally randomized controlled trials, in the evaluation of prevention strategies in pediatric sport. The integration of basic science, laboratory and epidemiological research is critical in evaluating the mechanisms associated with injury and injury prevention in pediatric sport. Finally, long-term studies are needed to identify the public health impact of pediatric sport injury. PMID- 16247259 TI - The Epidemiology of Children's Team Sports Injuries. PMID- 16247260 TI - Baseball injuries. AB - OBJECTIVE: To identify the frequency of injury in youth baseball, risk factors for these injuries, and possible prevention measures to reduce the frequency or severity of these injuries. DATA SOURCES: Information was collected from all known epidemiologic and intervention studies published in the peer-reviewed medical and scientific literature as it applies to youth baseball injuries. MAIN RESULTS: The frequency and severity of youth baseball injuries have remained relatively consistent over time. Risk factors for many injuries have been understudied and the study designs used for much of this research have been less than optimal. Several effective prevention measures have been identified, such as batting helmets, face shields, softer baseballs, and breakaway bases. CONCLUSIONS: Baseball is a relatively safe sport compared to many contact sports, but injuries do still occur. Future research should focus on determining the optimum pitching motion for both arm safety and performance, as well as systematically studying other potential safety improvements such as restrictions against breaking pitches. PMID- 16247261 TI - Basketball injuries. AB - OBJECTIVE: To identify and quantify, to the best extent possible from the existing literature, injury characteristics and factors (risk; protective) associated with injury in young basketball players. DATA SOURCES: Database searches principally involving Medline and SportDiscus. In addition, web-based searching and filtering of the reference lists of papers found in the preliminary searches were utilized. MAIN RESULTS: Few well-controlled studies of this population have been conducted. However, from the information available: basketball is the most frequent cause of sports-related emergency department visits for youth and adolescents; the risk of being injured in a game is greater than for practice; girls are more likely to be injured than boys, especially with knee and ankle injuries and the knee injuries are more likely to be severe; acute injuries are more common than chronic; strains/sprains are the most common types of injuries but overall time loss is minimal, indicating that the majority of pediatric basketball injuries are minor (less than 7 days away from activity). Intervention studies show that: mouthguards reduce orofacial/dental injuries; mouthguard use can be increased in young players; neuromuscular training can reduce the incidence of knee injuries in female participants; postural sway is related to risk of ankle injury. CONCLUSIONS: The current state of epidemiological research involving youth and adolescent basketball injuries is poor. With an increasing number of young participants, in situations ranging from informal play and physical education classes to organized community and school teams, the need for comprehensive and authoritative information on risk and protective factors is significant. PMID- 16247262 TI - Gridiron football injuries. AB - OBJECTIVE: To review the available football epidemiology literature to identify risk factors, facilitate injury prevention and uncover deficiencies that may be addressed by future research. DATA SOURCES: A literature search of Sports Discus (1940-2003), Eric (1967-2003), EMBASE (1988-2003), MEDLINE (1966-2003), CINAHL (1984-2003), and Web of Science (1993-2003) identified the published articles on American football in athletes of high school age and younger. MAIN RESULTS: Injury rate increases with the level of play (grade in school), player age, and player experience. The lower extremity (knee and ankle joints) is most frequently injured. Football injuries are much more common in games than in practice, and occur to players who are being tackled, tackling or blocking. Most injuries are mild, including contusion, strain and sprain. Rule changes with the prohibition of initial contact with the helmet or face-mask reduced catastrophic head and neck injuries. CONCLUSION: Although no sport or recreational activity is completely risk-free, football epidemiology research is critical to injury prevention. The existing medical literature provides some valuable insights, but an increased emphasis on prospective research is required to test the efficacy of preventative measures. Quality research may contribute to a reduction in football injury risk by defining the role of player conditioning and strength training, coaching of safety fundamentals, avoidance of dangerous activities, as well as proper medical supervision and care. Sports medicine personnel, coaches, and officials must strive to minimize injuries through progressive education, improved coaching techniques, effective officiating, and equipment modifications. PMID- 16247263 TI - Ice hockey injuries. AB - OBJECTIVE: This article reviews the distribution and determinants of injuries reported in the pediatric ice hockey literature, and suggests potential injury prevention strategies and directions for further research. DATA SOURCES: Thirteen electronic databases, the ISI Web of Science, and 'grey literature' databases were searched using a combination of Medical Subject Headings and text words to identify potentially relevant articles. The bibliographies of selected studies were searched to identify additional articles. Studies were selected for review based on predetermined inclusion and exclusion criteria. MAIN RESULTS: A comparison between studies on this topic area was difficult due to the variability in research designs, definition of injury, study populations, and measurements used to assess injury. The majority of injuries were sustained during games compared with practices. The two most commonly reported injuries were sprains/strains and contusions. Players competing at the Minor hockey, High School, and Junior levels of competition sustained most of their injuries to the upper extremity, head, and lower extremity, respectively. The primary mechanism of injury was body checking, followed by stick and puck contact. The frequency of catastrophic eye injuries has been significantly reduced with the world-wide mandation of full facial protection for all Minor hockey players. CONCLUSIONS: Specific hockey-related injury risk factors are poorly delineated and rarely studied among pediatric ice hockey players leaving large gaps in the knowledge of appropriate prevention strategies. Risk management strategies should be focused at avoiding unnecessary foreseeable risk, and controlling the risks inherent to the sport. Suggestions for injury prevention and future research are discussed. PMID- 16247265 TI - Soccer injuries. AB - OBJECTIVE: This chapter reviews the existing epidemiological studies on pediatric soccer injuries and discusses possibilities for future research. DATA SOURCES: A comprehensive, web-based search of existing soccer injury literature was performed with an emphasis on the pediatric population. The search encompassed all available studies, including European journals and texts, and initial investigations from the 1970s which serve as a basis of comparison to more recent work. MAIN RESULTS: Youth soccer is a relatively safe sport with an injury incidence ranging from 2.3 per 1,000 practice hours to 14.8 per 1,000 game hours. Similar to adults, youth soccer injuries occur mostly in the lower extremities, specifically the knee and ankle. Contusions are the most common injury, and minor/moderate injuries predominate. Extrinsic risk factors for youth soccer include: dangerous play, play on small fields, and inclusion of youth players on adult teams. The most important intrinsic risk factor is the relation of knee injury and female gender. CONCLUSIONS: Adolescent females suffer a disproportionate number of knee and anterior cruciate ligament injuries compared to adolescent males, but recent injury prevention studies yielded encouraging results. Head injuries in youth soccer are low, and rarely, if ever, occur from head to ball contact. Adherence to the rules of the game, proper coaching, and adequate refereeing are important factors in youth soccer injury prevention. PMID- 16247264 TI - Rugby injuries. AB - OBJECTIVES: The purpose of this chapter is to review critically the existing studies on the epidemiology of pediatric rugby injuries and discuss suggestions for injury prevention and further research. DATA SOURCES: Data were sourced from the sports medicine and science literature mainly since 1990, and from a prospective injury surveillance project in rugby undertaken by the University of New South Wales (UNSW) in Sydney during 2002. Literature searches were performed using Medline and SportsDiscus. MAIN RESULTS: Reported injury rates were between 7 and 18 injuries per 1,000 hours played, with the rate of injuries resulting in loss of playing or training time measured at 6.5-10.6 per 1,000 hours played. Injury rates increased with age and level of qualification. Head injury and concussion accounted for 10-40% of all injuries. In the UNSW study, concussion accounted for 25% of injuries resulting in loss of playing or training time in the under 13 year age group. Upper and lower extremity injuries were equally apportioned, with musculoskeletal injuries being the main type of injury. Fractures were observed in the upper extremity and ankle, and joint/ligament injuries affected the shoulder, knee and ankle. The tackle was associated with around 50% of all injuries. The scrum produced fewer injuries, but is historically associated with spinal cord injury. CONCLUSIONS: Rugby is a contact sport with injury risks related to physical contact, primarily in the tackle. Most injuries affect the musculoskeletal system, with the exception of concussion. Spinal cord injury is rare, but catastrophic. Research is required to understand better injury risks and to reduce the incidence of shoulder, knee and ankle joint injuries, concussion and spinal injury. PMID- 16247266 TI - Injury prevention and future research. AB - OBJECTIVES: To critically examine and summarize the literature identifying risk factors and prevention strategies for injury in child and adolescent sport. DATA SOURCES: Seven electronic databases were searched including: Medline, Cumulative Index to Nursing and Allied Health Literature (CINAHL), Psychinfo, Cochrane Database for Systematic and Complete Reviews, Cochrane Controlled Trials Registry, HealthSTAR and SPORTDiscus. Medical subject headings and text words included: athletic injury, sport injury, risk factors, adolescent and child. Additional articles were reviewed based on sport-specific contributions in the previous chapters of this book. MAIN RESULTS: Despite the diversity of injuries occurring in various pediatric sporting populations, the uniformity with respect to many of the risk factors identified in the literature is noteworthy (i.e. previous injury, age, sport specificity, psychosocial factors, decreased strength and endurance). The literature is significantly limited with respect to the prospective evaluation of risk factors and prevention strategies for injury in pediatric sport. The consistencies, however, between the adult and pediatric literature are encouraging with respect to prevention strategies involving neuromuscular training programs (i.e. balance training programs) to reduce lower extremity injuries in some sports and the use of sport-specific protective equipment (i.e. helmets). CONCLUSIONS: Notwithstanding the limitations in the literature, the successful evaluation of some sport-specific prevention strategies to reduce injury in pediatric sport is encouraging. There is significant opportunity to methodologically improve upon the current pediatric sport injury literature in descriptive surveillance research, risk factor evaluation research, and prevention research. There is a need for prospective studies, ideally randomized controlled trials, in the evaluation of prevention strategies in pediatric sport. The integration of basic science, laboratory and epidemiological research is critical in evaluating the mechanisms associated with injury and injury prevention in pediatric sport. Finally, long-term studies are needed to identify the public health impact of pediatric sport injury. PMID- 16247267 TI - [A case of abdominal mesenteric aneurysm in a young woman]. PMID- 16247268 TI - [The relationship between ineffective esophageal motility and gastro-esophageal reflux disease]. AB - BACKGROUND/AIMS: Ineffective esophageal motility (IEM) is a distinct manometric entity characterized by a hypocontractile esophagus. Recently, IEM replaced the nonspecific esophageal motility disorder (NEMD), and its associations with gastro esophageal reflux disease (GERD) and respiratory symptoms are well known. We evaluated the relationship of IEM with GERD, and the diagnostic value of IEM for GERD. METHODS: We retrospectively analyzed recent 3-year (Jan. 1998-Sep. 2002) datas of esophageal manometry, acid perfusion test and simultaneous 24 hr ambulatory pH-metry with manometry studies in 270 consecutive patients with esophageal and/or GERD symptoms. The prevalence of IEM in GERD group and non-GERD group, and the variables of pH-metry and manometry among esophageal motility disorders were compared. In addition, the sensitivity, specificity, positive predictive value, negative predictive value of IEM, esophageal symptom, and acid perfusion test for GERD were calculated. RESULTS: There was no significant difference in IEM prevalence rate between GERD group and non-GERD group. In addition, there was no significant difference in GERD prevalence rate and esophageal acid clearance in variety of motility disorder groups. Total percent time of pH <4 in IEM group did not show any difference when compared with other groups except in the achalasia group. In regard of diagnostic value to detect GERD, all positive results showed high specificity (97%) in IEM with esophageal symptom and positive acid perfusion test. CONCLUSIONS: The diagnosis of IEM using esophageal manometry in patients with various esophageal symptoms does not strongly suggest on association with GERD. However, IEM with concomitant esophageal symptoms and positive acid perfusion test has diagnostic values for GERD. PMID- 16247269 TI - [Clinical effectiveness of transcatheter arterial embolization for acute upper and lower non-variceal gastrointestinal bleeding]. AB - BACKGROUND/AIMS: Although the initial rate of hemostasis achieved by endoscopic treatment for acute non- variceal gastrointestinal bleeding (NVGIB) is high, recurrent or persistent bleeding occurs in 10% to 25% of the patients. The aim of this study was to assess the efficacy and safety of transcatheter arterial embolization (TAE) in patients with acute upper and lower NVGIB who could not be managed by endoscopic treatment. METHODS: A retrospective analysis of the clinical data was done in 43 patients (M/F: 26/17, mean age: 60 years) whom underwent angiography or TAE for acute upper and lower NVGIB between January 1998 and December 2003. Among 43 patients, 18 had upper NVGIB, 19 had lower NVGIB, and 6 had obscure gastrointestinal bleeding. Demographic characteristics and outcome parameters including the rates of hemostasis, in-hospital death, and complications were analyzed. RESULTS: Thirty-four patients underwent TAE while 9 patients underwent angiography. TAE was used as the first line treatment in 17 patients and as the second line treatment in others. Hemostasis was achieved in 29 of 34 patients (85.3%) by TAE. According to the site of bleeding, hemostasis was achieved in 14 of 17 patients (82.4%) with upper NVGIB and in 15 of 17 patients (88.2%) with lower NVGIB. There was no significant angiography or TAE related complications such as bowel ischemia or infarction except a hematoma on the angiography site in one patient. CONCLUSIONS: TAE is effective and safe in patients with acute upper or lower NVGIB who cannot be managed by endoscopic treatment. PMID- 16247270 TI - [Grading of histology, expression of apoptosis and cell proliferation in gastric mucosa adjacent to gastric adenoma or adenocarcinoma]. AB - BACKGROUND/AIMS: Helicobacter pylori (H. pylori) infection can lead to gastric adenoma and carcinoma through atrophic gastritis and intestinal metaplasia. Imbalance between apoptosis and proliferation may play a role in gastric carcinogenesis. We tried to investigate H. pylori infection rate, grade of gastritis, environmental risk factors, expression rate of apoptosis and cell proliferation in mucosa adjacent to tumor, and we also tried to find significant factors associated with gastric carcinogenesis. METHODS: Endoscopically diagnosed twenty cases of intestinal type gastric carcinoma, 20 cases of gastric adenoma, and 40 cases of control (normal or gastritis) were enrolled. H. pylori infection rate, histologic grading, apoptosis and immunohistochemical stain (Ki-67 and p53) to check mucosal proliferation were done in endoscopically biopsied tissues at antrum and body at least 2 cm apart from adenoma or carcinoma. RESULTS: In three groups, H. pylori infection rates were not significantly different. In the multivariate analysis, only atrophy of gland was a significant risk factor for adenoma compared to control group (OR 3.7). Intestinal metaplasia in antrum and alcohol drinking were significant risk factors for carcinoma compared to control group (OR 4.4 and 4.9 respectively). Expressions of apoptosis, Ki-67 and p53 were not significantly different in three groups. CONCLUSIONS: Intestinal metaplasia in antrum and alcohol drinking are significant risk factors for gastric carcinoma. Degree of mucosal proliferation and apoptosis in gastric mucosa adjacent to tumor are not significantly different in three groups. PMID- 16247271 TI - [The current status of ulcerative colitis-associated colorectal cancer in Korea: a KASID study]. AB - BACKGROUND/AIMS: The number of patients with ulcerative colitis has steadily increased since the mid-1980s. As the risk of colon cancer increases with duration and extents, colitic cancers are expected to increase in number in Korea. We surveyed the current status of colitic cancers and provided the perspectives in Korea. METHODS: Korean Association for the Study of Intestinal Diseases (KASID) collected and descriptively analyzed a total of eleven cases of colitic cancers occurred between 1993 and 2003. RESULTS: The mean age at the diagnosis of colitic cancer was 49.3 years, and the mean duration of ulcerative colitis was 12.5 years. Of the diagnosed cases, 91% was associated with pancolitis, 28% had right colon cancers and 18.2% had multiple tumors. Colitic cancers diagnosed at Dukes' stage D were 36.3%. Stages at diagnosis were lower in patients with good compliance. Up to 2003, cumulative incidence ratio of colitic cancer to ulcerative colitis was estimated to be 0.5%, which was much less than the overall prevalence of 3.7% in the Western countries. This incidence was also less than the expected cumulative incidence ratio of 0.9%, which was estimated by adopting the average results of meta-analysis in the Western world. CONCLUSIONS: As the incidence of ulcerative colitis has not reached a plateau, and as the diagnosis of colitic caners are delayed in Korea, the cumulative incidence of colitic cancers might have appeared to be low. Geographic and racial factors, and the effect of preventive measures might have contributed to the lower incidence which remains to be proven. Since colitic cancers are expected to increase steeply in the near future in Korea, the importance of surveillance and prevention cannot be over-emphasized. PMID- 16247272 TI - [Prognostic factors and survival according to the Okuda stage in patients with hepatocellular carcinoma]. AB - BACKGROUND/AIMS: There are several staging systems to decide the stage of hepatocellular carcinoma (HCC), but yet incomplete. Okuda stage which includes both tumor characteristics and liver function is widely used. The aims of this study were to assess the usefulness of known prognostic factors and Okuda staging system in 237 cases of HCC. METHODS: A retrospective analysis of 237 cases of HCC diagnosed from 2000 to 2002 was performed. We analyzed prognostic factors such as age, sex, liver cirrhosis, Child-Pugh classification, tumor size, albumin, bilirubin, alpha-FP, ascites, encephalopathy and Okuda stage. Prognostic analysis was performed for single variables and estimating survival distributions were analyzed by the Kaplan-Meier method, statistically compared by the log-rank test. RESULTS: Patients had a mean age of 57.5 years and were predominantly men (79.7%). Liver cirrhosis were noticed in 214 cases (90.3%). The overall median survival period was 25.7 months. The median survival period was correlated to bilirubin, ascites, alpha-FP, tumor size, and Child-Pugh classification, but not to age, sex, and pattern of viral infection. The median survival period of the Okuda stage I, II and III cases was 35.8, 11.9 and 8.5 months (p<0.001). CONCLUSIONS: The median survival period of patients with HCC is significantly correlated to Okuda staging system, and survival period has improved than the initial data when the Okuda staging system was published in 1985. However, in order to discriminate early staged HCC more accurately, other prognostic factors such as alpha-FP and tumor morphology should be included in future staging system for HCC. PMID- 16247273 TI - [Expression of hedgehog proteins in periampullary cancer]. AB - BACKGROUND/AIMS: Hedgehog protein is an essential molecule for gastrointestinal tract development, and disruption of hedgehog signaling pathway is linked to some gastrointestinal tumorigenesis. Here, we performed hedgehog immunostaining in periampullary cancer to evaluate the differences according to the location type of cancer and the differentiation of adenocarcinoma. METHODS: We retrieved surgical specimens from 43 periampullary cancer patients (15 ampulla of Vater cancer, 12 distal common bile duct cancer, 13 pancreatic head cancer, and 3 combined ampulla of Vater/bile duct cancer). Immunohistochemical stain was performed in both normal and cancerous tissue portions of each case using Sonic hedgehog (H-160) rabbit polyclonal antibody. Immunohistochemical stain results were grouped into three groups according to the percentage of positive cytoplasmic stain in tumor volume (unstained: <5%, weakly stained: 5-50%, and strongly stained: >50%). RESULTS: All of the normal tissue revealed negative immunohistochemical stain while cancerous tissue revealed positivity in 95.3% (41/43 cases). Strongly stained cases were more frequently seen in ampulla of Vater cancers (13/15) and in combined ampulla of Vater/bile duct cancers (3/3) than in distal common bile duct cancers (4/12) and in pancreatic head cancers (3/13) (p=0.002). In addition, strongly stained cases were more frequently seen in well-differentiated adenocarcinoma than the others (p<0.001). CONCLUSIONS: Most of the periampullary cancers show hedgehog protein expression. In addition, hedgehog protein immunostainings shows stronger expression in ampulla of Vater cancers and in well-differentiated adenocarcinoma. PMID- 16247274 TI - [A case of refractory pediatric Crohn's disease with a successful treatment by infliximab therapy]. AB - Crohn's disease is a chronic inflammatory bowel disease which affects mainly children and young adults, and its cause remains unknown so far. Infliximab, a monoclonal antibody to the pivotal cytokine tumor necrosis factor-alpha, has been approved as a drug for both induction and maintenance therapy for moderately to severely active, or fistula-complicated Crohn's disease. The authors report a 12 year-old male patient diagnosed as Crohn's disease complicated with a perianal fistula, which was refractory to the conventional therapy. After the 0, 2, and 6 week scheduled intravenous infusion of infliximab, the patient reached to clinical remission in both subjective symptoms and objective manifestations. For children or young adults who develop Crohn's disease in a refractory course, infliximab may serve as a drug which leads to a clinical improvement or even to an extent of remission. PMID- 16247275 TI - [A case of retroperitoneal schwannoma of the vagus nerve]. AB - Schwannomas are benign nerve sheath tumors that originate from any anatomical site. Most schwannomas occur in the head, neck or limbs, but rarely occur in the retroperitoneal space. Furthermore, the schwannoma originating from the vagus nerve of retroperitoneal space is much rare. We experienced a case of retroperitoneal schwannoma of the vagus nerve. A 34-year-old male was referred to our hospital for the evaluation of abdominal mass on ultrasonography. Endoscopic examination revealed submucosal tumor-like lesion on high body of the stomach. Computed tomography (CT) revealed that the stomach was compressed by a solid tumor in the retroperitoneum. On exploratory laparotomy, this mass turned out to be a baseball sized mass in the retroperitoneal space. The mass was excised in an encapsulated state. Histological examination with immunohistochemical stains revealed a schwannoma of the vagus nerve. PMID- 16247276 TI - [A common bile duct web in association with common bile duct stone]. AB - Webs are diagnosed by their characteristic appearance on imaging studies, typically appearing as thin, radiolucent rings with or without dilatation of the organ proximal to it. Like in other organs, the etiology of webs in the common bile duct is controversial. Some webs are thought to be congenital, whereas others occur in the presence of chronic inflammation, suggesting a pathogenic relationship. We report a case of a common bile duct septum in association with numerous large black pigment stones in a 62-year-old woman. The patient was treated by cholecystectomy with T-tube insertion. PMID- 16247277 TI - [Prognosis and staging system of hepatocellular carcinoma]. PMID- 16247279 TI - A focus on heart failure and other age-related cardiovascular conditions. PMID- 16247280 TI - Does increased arterial stiffness increase the risk for postural hypotension? AB - This study tests the hypothesis that increased arterial stiffness is associated with postural hypotension in older adults. Aortic pulse wave velocity and postural blood pressure (BP) response were assessed in 49 nondiabetic community dwelling normotensive (n=27) and hypertensive (n=22) older adults (mean age+/-SD, 71+/-6.7 years) who were not receiving vasoactive medications. During the 5 minute period of upright posture, 13 subjects had no change or a postural increase in systolic BP (SBP)(+10.6+/-14.6 mm Hg), 27 had a postural decrease of <20 mm Hg (-9.3+/-4.2 mm Hg), and nine had a postural decrease of >20 mm Hg ( 29.1+/-8.1 mm Hg). Contrary to the proposed hypothesis, pulse wave velocity was significantly greater in subjects with a postural increase in SBP than in those with a postural decrease in SBP<20 mm Hg (10.2+/-0.68 m/sec vs. 8.3+/-0.37 m/sec; p=0.03) and tended to be greater than in those with a postural decrease in SBP>20 mm Hg (10.2+/-0.68 m/s vs. 8.5+/-0.73 m/sec; p=0.11). Higher pulse wave velocity was associated with a more positive postural SBP response at 1 minute (r=0.42; p=0.024), 3 minutes (r=0.38; p=0.007), and 5 minutes (r=0.45; p=0.001). This study does not support a relationship between arterial stiffness and a postural decrease in BP among healthy older adults; other age-related factors regulating BP homeostasis likely play a greater role. PMID- 16247281 TI - Titration of carvedilol in elderly heart failure patients. AB - Physicians may withhold beta blockers from elderly heart failure patients based on perceptions of poor tolerability in this group. This study analyzed the success and tolerability of carvedilol titration in elderly (70 years and older) and younger (younger than 70) heart failure patients (n=360). The mean duration of titration was similar in both groups. The target carvedilol dosage (50 mg/d) was achieved by 55.3% and 62.0%, and a partial dose was achieved by 44.7% and 35.5% (p=0.028) of elderly and younger patients, respectively. The mean achieved dosage was lower in elderly patients than in younger patients (42.4+/-27.2 mg/d vs. 55.1+/-30.1 mg/d; p<0.0001), possibly due to significant between-group differences in baseline characteristics or rates of target dose achieved. When adjusted for body weight, the mean achieved dose was similar in elderly (0.58+/ 0.32 mg/kg/d) and younger (0.64+/-0.30 mg/kg/d) patients (p=nonsignificant). Adverse events were more frequent in the elderly but did not appear to significantly impact titration duration or carvedilol dose achieved when corrected for body weight. In conclusion, carvedilol should not be withheld from elderly heart failure patients based on perceptions of poor potential tolerability. PMID- 16247282 TI - Ambulatory hemodynamics in patients with chronic heart failure: implications for volume management in elderly patients. AB - Invasive hemodynamic assessment of patients with chronic heart failure is helpful in managing severe congestive symptoms associated with acute volume exacerbations that lead to hospitalizations. Information useful in guiding medication changes targeting control of left ventricular filling pressures can be obtained from measurements in the right ventricle. With the advent of implantable devices designed to treat heart failure, the logical next development is an implanted device that incorporates sophisticated monitoring systems capable of continuous acquisition of hemodynamic information over a long-term period. An implantable hemodynamic monitoring system (Chronicle; Medtronic, Inc., Minneapolis, MN) is safe to implant and has a pressure sensor with proven long-term reliability. Using transtelephonic data transmission and Internet-based information systems, ambulatory hemodynamic monitoring may reduce the need for hospitalizations and overall health care utilization in patients with symptomatic heart failure. This approach may be especially helpful for elderly patients with chronic heart failure in whom overdiuresis or volume contraction may lead to gait instability and falls. Additionally, elderly patients with heart failure from primary diastolic dysfunction may have a very narrow therapeutic window of optimal volume that is difficult to assess by daily weights and physical examination alone. This hypothesis is currently being tested in a prospective randomized clinical trial. PMID- 16247283 TI - Atrial fibrillation: management for the geriatric patient. AB - Atrial fibrillation is one of the most common rhythm disorders encountered in the geriatric population. The diagnosis is well established, but the treatment has undergone many changes. Many long-held notions have been challenged and new treatment options, both medical and procedural, have offered new hope in the management of this disease in the elderly. This review will describe atrial fibrillation from its diagnosis to the common and more cutting-edge treatment modalities with special focus on the elderly population. PMID- 16247284 TI - Third pivotal research in cardiology in the elderly (PRICE-III) symposium: heart failure in the elderly: mechanisms and management. PMID- 16247285 TI - Is postural adaptation impaired by hardened arteries? PMID- 16247286 TI - Ethical issues in the management of geriatric cardiac patients. Special report: Terri's Tale: dispelling the myths in the Terri Schiavo Ordeal. PMID- 16247288 TI - Choice of pacemaker mode in high-grade heart block. PMID- 16247287 TI - Papillary fibroelastoma: an uncommon cause for a transient ischemic attack. PMID- 16247289 TI - Hypocalcemia, hyperkalemia, and junctional or sinoventricular rhythm. PMID- 16247290 TI - Economic analyses of human genetics services: a systematic review. AB - PURPOSE: The study's purpose was to conduct a structured review of economic analyses of genetic services. These will be increasingly valuable tools for assessing the clinical and economic outcomes of new medical technologies. METHODS: We searched for economic studies published between January 1990 and August 2004 from a variety of publicly available databases. Articles were first reviewed to determine whether they were original studies, and second to determine whether they were formal cost-effectiveness analyses by established criteria. Articles meeting these criteria were graded using a validated rating scale. RESULTS: Of 149 articles, 63 met established criteria for cost-effectiveness analyses. The majority (87%) were published since 1996. The majority of studies considered adult (31) or prenatal (25) conditions with the remainder considering preconception or pediatric conditions. More than half used life years gained or an ad hoc measure of outcome (e.g., cases detected). Twenty-five percent measured outcome as quality-adjusted life years. The disease areas most considered were cancer (21%) and aneuploidies (18%). The average quality ranking was 87 of 100 possible (range 48-100). Common shortcomings included lack of statement of perspective, lack of discussion of potential bias, and lack of disclosure of funding sources. CONCLUSIONS: Relatively few economic evaluations are available for genetic services, and most are clustered in specific disease areas. Overall quality was high, but varied widely. Most shortcomings that would improve study quality are easy to address. To improve the relevance of these studies, researchers need to incorporate measures of outcome that are familiar to decision makers, including quality-adjusted life years. PMID- 16247291 TI - NSD1 analysis for Sotos syndrome: insights and perspectives from the clinical laboratory. AB - PURPOSE: Sotos syndrome is a genetic disorder characterized primarily by overgrowth, developmental delay, and a characteristic facial gestalt. Defects in the NSD1 gene are present in approximately 80% of patients with Sotos syndrome. The goal of this study was to determine the incidence of NSD1 abnormalities in patients referred to a clinical laboratory for testing and to identify clinical criteria that distinguish between patients with and without NSD1 abnormalities. METHODS: Deletion or mutation analysis of the NSD1 gene was performed on 435 patients referred to our clinical genetics laboratory. Detailed clinical information was obtained on 86 patients with and without NSD1 abnormalities, and a clinical checklist was developed to help distinguish between these two groups of patients. RESULTS: Abnormalities of the NSD1 gene were identified in 55 patients, including 9 deletions and 46 mutations. Thus, in the clinical laboratory setting, deletions were found in 2% and mutations in 21% of samples analyzed, because not all patients had both tests. Thirty-three previously unreported mutations in the NSD1 gene were identified. Clinical features typically associated with Sotos syndrome were not found to be significantly different between individuals with and without NSD1 abnormalities. The clinical checklist developed included poor feeding, increased body mass index, and enlarged cerebral ventricles, in addition to the typical clinical features of Sotos syndrome, and was able to distinguish between the two groups with 80% sensitivity and 70% specificity. CONCLUSIONS: The dramatic decrease in the frequency of finding NSD1 abnormalities in the clinical laboratory is likely because of the heterogeneity of the patient population. Our experience from a diagnostic laboratory can help guide clinicians in deciding for whom NSD1 genetic analysis is indicated. PMID- 16247293 TI - Comparison of genotypic and phenotypic strategies for population screening in hemochromatosis: assessment of anxiety, depression, and perception of health. AB - PURPOSE: Hemochromatosis is a treatable disorder with a major genetic predisposition. It provides an example in which genotypic and phenotypic strategies for screening may be compared. We previously showed noninferiority of uptake of a genotypic population screening strategy for hemochromatosis compared with a phenotypic strategy. In this article we present the psychologic effects of each strategy. METHODS: A sample of 3000 individuals from primary care were randomly allocated to a phenotypic or genotypic screening strategy for hemochromatosis, and the 939 individuals who accepted screening provide the sample for this article. Standardized assessments of anxiety, general health, and depression were made at invitation, testing, result-giving, and 6 months. RESULTS: Screening did not lead to significant changes in the self-rated assessments of anxiety, depression, and general health over time, and there were no significant differences between the two screening strategies. The unemployed or permanently disabled had lower ratings of health and higher anxiety and depression. CONCLUSION: The two screening strategies appeared to cause little adverse psychologic disturbance in the short term, and there was no difference between the two strategies This study provides some empiric data to support arguments against "genetic exceptionalism" and suggests that genetic testing when used for population screening for a treatable disease has few adverse effects. PMID- 16247292 TI - Developing a sustainable process to provide quality control materials for genetic testing. AB - PURPOSE: To provide a summary of the outcomes of two working conferences organized by the Centers for Disease Control and Prevention (CDC), to develop recommendations for practical, sustainable mechanisms to make quality control (QC) materials available to the genetic testing community. METHODS: Participants were selected to include experts in genetic testing and molecular diagnostics from professional organizations, government agencies, industry, laboratories, academic institutions, cell repositories, and proficiency testing (PT)/external Quality Assessment (EQA) programs. Current efforts to develop QC materials for genetic tests were reviewed; key issues and areas of need were identified; and workgroups were formed to address each area of need and to formulate recommendations and next steps. RESULTS: Recommendations were developed toward establishing a sustainable process to improve the availability of appropriate QC materials for genetic testing, with an emphasis on molecular genetic testing as an initial step. CONCLUSIONS: Improving the availability of appropriate QC materials is of critical importance for assuring the quality of genetic testing, enhancing performance evaluation and PT/EQA programs, and facilitating new test development. To meet the needs of the rapidly expanding capacity of genetic testing in clinical and public health settings, a comprehensive, coordinated program should be developed. A Genetic Testing Quality Control Materials Program has therefore been established by CDC in March 2005 to serve these needs. PMID- 16247295 TI - Newborn blood spot screening and genetic services: a survey of Minnesota primary care physicians. AB - PURPOSE: To (1) obtain guidance on the preferred content and format of quick reference newborn blood spot screening information from the Minnesota Department of Health; (2) determine primary care physicians' perceptions of the benefits of genetic services; and (3) determine primary care physicians' satisfaction with genetic counseling services. METHODS: A written survey was mailed to family physicians and pediatricians in Minnesota (n = 300). RESULTS: Eighty physicians responded (28% response rate). Whereas 70% of respondents felt previous information received from the newborn screening program was adequate, 83% were interested in quick reference information. The majority of physicians preferred this information as a laminated sheet (63%). Physician procedure for an abnormal screen, newborn screening program protocol for an abnormal screen, and disease treatment and follow-up information were recommended for inclusion on quick reference. Over half of physicians agreed with the following benefits of genetic services: provide testing options (88%); evaluate family members (88%); reduce parental anxiety (87%); provide resources (83%); provide diagnostic information (76%); determine medical needs (67%); and determine emotional needs (51%). Ninety nine percent of physicians were satisfied with genetic counseling services. CONCLUSIONS: Physicians indicated that reference material for primary care physicians should include a quick reference card with specific categories of information. Newborn screening programs should attempt to increase physician awareness of genetic services, including the subsequent medical and psychosocial benefits for their patients. PMID- 16247294 TI - Patient acceptability of genotypic testing for hemochromatosis in primary care. AB - PURPOSE: Genetic screening can enable timely detection and treatment of hereditary hemochromatosis (HH). Little is known about patient acceptability of DNA testing as compared to conventional phenotypic testing. METHODS: Within the HEIRS Study, a large primary-care screening study of HH and iron overload, we randomly assigned participants to receive brief information on either HH genotypic or phenotypic testing, and assessed the willingness to accept this test. The study was designed to recruit an equal number of African Americans and Caucasians. RESULTS: A total of 2500 participants were recruited from waiting rooms of primary care practices; 2165 participants who self-identified as African Americans and Caucasians were included in the analyses. Overall, 56% had accepted a genotypic test versus 58% for a phenotypic test. Adjusting for Field Center (FC), age, gender, race, educational attainment, global health rating, and knowledge of the test, the odds ratio of accepting a genotypic versus phenotypic test was 0.85 (95% CI: 0.71, 1.02; P = 0.078). Characteristics associated with test acceptance were age 45-64 years, female gender, Caucasian race, self-rated health less than ''very good'', and knowledge of the test. Test acceptance was associated with interest in knowing more about health (81%) and in helping family members (71%). Refusal reasons included a need to talk with a doctor (44%), concern about privacy (32%), and dislike of blood drawing (29%). CONCLUSION: In this diverse sample of primary care patients, stated acceptance of genotypic testing for HH mutations was similar to phenotypic testing for blood iron. Patient education regarding the nature of test, importance of disease detection, and privacy protection appear to be essential for achieving high rates of screening participation. PMID- 16247296 TI - Technical standards and guidelines: molecular genetic testing for ultra-rare disorders. AB - These standards and guidelines are designed primarily as an educational resource for clinical laboratory geneticists to help them provide quality clinical laboratory genetic services. Adherence to these standards and guidelines does not necessarily ensure a successful medical outcome. These standards and guidelines should not be considered inclusive of all proper procedures and tests or exclusive of other procedures and tests that are reasonably directed to obtaining the same results. In determining the propriety of any specific procedure or test, the clinical molecular geneticist should apply his or her own professional judgment to the specific clinical circumstances presented by the individual patient or specimen. It may be prudent, however, to document in the laboratory record the rationale for any significant deviation from these standards and guidelines. PMID- 16247297 TI - Fragile X syndrome: diagnostic and carrier testing. AB - This guideline is designed primarily as an educational resource for medical geneticists and other health care providers to help them provide quality medical genetic services. Adherence to this guideline does not necessarily assure a successful medical outcome. This guideline should not be considered inclusive of all proper procedures and tests or exclusive of other procedures and tests that are reasonably directed to obtaining the same results. In determining the propriety of any specific procedure or test, the geneticist should apply his or her own professional judgment to the specific clinical circumstances presented by the individual patient or specimen. It may be prudent, however, to document in the patient's record the rationale for any significant deviation from this guideline. PMID- 16247298 TI - Peas, priorities and primary care: cultural competency starts at home. PMID- 16247299 TI - In defense of commercial laboratories. PMID- 16247301 TI - Musculoskeletal trauma care means orthopaedic trauma care. PMID- 16247302 TI - Contaminant seeding in bone by different irrigation methods: an experimental study. AB - OBJECTIVE: This study was designed to investigate the effectiveness of using various devices and manual procedures for cleansing bacterially contaminated bone tissue and to assess the risk of iatrogenic bacterial seeding in deep bone layers. METHODS: In an in vitro model, human femoral heads were contaminated with Escherichia coli and then cleansed with pulsatile high-pressure lavage, pulsatile low-pressure lavage, manual rinsing with bulb syringe lavage, or manual rinsing with combined brush cleaning. The numbers of bacteria that remained or those that were introduced by the rinsing procedures were quantitatively determined at depths of 0 to 1 cm, 1 to 2 cm, and 2 to 3 cm. RESULTS: Both pulsatile high pressure lavage and brush cleaning were more effective than pulsatile low pressure lavage and bulb syringe lavage for the purpose of surface cleansing. The differences were highly significant (P < 0.001). There was no significant difference in the decontaminating effect between pulsatile high-pressure lavage and brush cleaning (P = 0.24). The bacterial contamination attributable to the cleansing procedure, as measured at tissue depths of 1 to 2 cm and 2 to 3 cm, was significantly higher after pulsatile high-pressure lavage and after pulsatile low pressure lavage than it was after bulb syringe lavage or brush cleaning (P < 0.001). CONCLUSION: In this in vitro investigation of cancellous bone, the brush cleansing was just as effective for getting rid of bacterial contamination as pulsatile high-pressure lavage, and carries a significantly lesser risk of iatrogenic bacterial seeding into deeper tissue layers. In the light of these promising results obtained by the cleansing of cancellous bone contaminated with bacteria, it would be desirable to perform supplementary in vitro and in vivo investigations into brush cleansing. PMID- 16247303 TI - The mechanical behavior of locking compression plates compared with dynamic compression plates in a cadaver radius model. AB - OBJECTIVE: The purpose of this cadaveric study was to compare the mechanical behavior of a locked compression plate, which uses threaded screw heads to create a fixed angle construct, with a dynamic compression plate construct in a cadaver radius model. DESIGN: Mechanical study with cyclic testing and high-speed optical motion analysis. SETTING: Biomechanics laboratory at an academic institution. PATIENTS/PARTICIPANTS: Eighteen pairs of fresh-frozen human cadaver radii were divided into 3 groups of 6 to be tested as a group in each of the following force applications: anteroposterior (AP) bending, mediolateral bending, or torsion. INTERVENTION: Each bone was osteotomized leaving a 5-mm fracture gap and then fixed with a plate. For each pair, 1 radius received a standard plate (limited contact dynamic compression plates; LC-DCP), the contralateral radius was fixed with a locking compression plate (LCP), and specimens underwent cyclic loading. Normalized stiffness, average energy absorbed, and Newton-cycles to failure were calculated. In addition, a 3-dimensional, high-speed, infrared motion analysis system was used to evaluate motion at the fracture site. MAIN OUTCOME MEASUREMENTS: Construct stiffness, fracture site motion, cycles to failure, and energy absorption. Repeated measures ANOVA were used to detect differences between groups with time. RESULTS: In the torsion group, LCP specimens failed at 60% greater Newton-cycles than the LC-DCP (1473 vs. 918; P < 0.05). In the AP group, the LC-DCP absorbed significantly greater energy during 10,000 cycles compared with the LCP group (P < 0.05). The 2 constructs demonstrated different biomechanical behavior with time. As cycling progressed in the LC-DCP specimens under torsion testing, stiffness (measured at the actuator at the bone ends) did not change significantly; however, fracture motion (measured at the fracture surfaces) decreased significantly (P = 0.04). The LCP specimens did not display similar behavior. CONCLUSIONS: Our findings indicated that LCP constructs may demonstrate subtle mechanical superiority compared with the LC-DCP. The LCP specimens had less energy absorption in the AP group and survived longer in the torsion group. Discordance of motion between measurement regions was observed only in the LC-DCP torsion group, and may have been caused by plate-bone slippage or bone-screw subcatastrophic failure. However, many other compared parameters were found to be similar, and the clinical significance of the few differences found between constructs mandates further investigation. PMID- 16247304 TI - Intraoperative diagnosis of syndesmosis injuries in external rotation ankle fractures. AB - OBJECTIVE: This study was designed to compare intraoperative fluoroscopic stress testing, static radiographs, and biomechanical criteria for the diagnosis of distal tibiofibular syndesmotic instability associated with external rotation type ankle fractures. DESIGN: Prospective, consecutive series. SETTING: Academic level 1 trauma center. PATIENTS/PARTICIPANTS: Thirty-eight skeletally mature patients with unstable unilateral external rotation ankle fractures were prospectively recruited. INTERVENTION: Before surgery, the treating surgeon detailed the operative treatment plan, including need for syndesmotic fixation. In pronation-external rotation injuries, biomechanical criteria were applied to predict syndesmotic instability. Ankles were examined using intraoperative fluoroscopic external rotation stress tests. The contralateral uninjured limb was used as a control. A 7.2-Nm force was applied for the external rotation stress examination. Stress testing was performed after lateral malleolar fixation and repeated after medial and syndesmotic fixation. MAIN OUTCOME MEASURES: The incidence of syndesmotic instability was determined based on radiographic clear space measurements and compared with previously published criteria. RESULTS: Intraoperative fluoroscopy detected unpredicted syndesmotic instability in 37% of ankles. In supination-external rotation (OTA 44B) injuries, unpredicted syndesmosis instability was found in 10 of 30 patients (33%). In pronation external rotation injuries (OTA 44C), 4 of 7 patients (57%) were associated with syndesmosis disruption not predicted by biomechanical criteria. In bimalleolar fractures, syndesmosis fixation improved stability compared with rigid bimalleolar fixation alone (P < 0.01). CONCLUSIONS: Preoperative radiographs and biomechanical criteria are unable to routinely predict the presence or absence of syndesmosis instability. Rigid bimalleolar fixation was frequently not sufficient to stabilize syndesmotic disruption. Intraoperative stress fluoroscopy is a valuable tool for detection of unstable syndesmotic injuries. PMID- 16247305 TI - Computer-assisted guidewire insertion for hip fracture fixation. AB - OBJECTIVE: : This study was designed to test in a laboratory setting a novel computer-assisted fluoroscopic technique and a conventional fluoroscopic technique for open reduction and internal fixation (ORIF) of hip fractures. Our hypothesis is that a novel computer-assisted fluoroscopic technique will achieve acceptable guidewire placement in one pass, with decreased fluoroscopic time and with accuracy and precision better than conventional technique. DESIGN: Prospective, randomized trials. SETTING: Laboratory. PARTICIPANTS: Thirty, Sawbone, femur phantoms. INTERVENTION: Dynamic hip screw guidewires were inserted into 15 femur phantoms under fluoroscopic guidance by using computer-assisted fluoroscopic ORIF technique, and 15 femurs were inserted by using a conventional fluoroscopic-assisted ORIF technique. MAIN OUTCOME MEASUREMENTS: Ideal guidewire placement was defined as the center of the femoral head, 5 mm from the apical bone edge on anteroposterior and lateral views. Accuracy was measured as distance to ideal placement, and the number of passes and fluoroscopic time were noted for each trial. RESULTS: The computer-assisted technique achieved an average guidewire placement that was as accurate as the conventional technique in fewer passes, 1.1 +/- 0.2 (mean +/- standard deviation) compared with 2.4 +/- 1.1 (P < 0.0001), respectively, and with fewer fluoroscopic images, 2 +/- 0 compared with 13.5 +/- 3 (P < 0.0002), respectively. Guidewire placement in both groups was within the tip-apex distance defined by Baumgaertner et al. CONCLUSIONS: The computer-assisted technique was significantly more accurate and precise than conventional technique. It also required fewer drill tracks through the femur and exposed the patient and the surgical team to significantly less ionizing radiation. PMID- 16247306 TI - Interobserver reliability of a CT-based fracture classification system. AB - OBJECTIVES: This study was designed to determine whether the interobserver reliability of a fracture classification scheme applied based on a single, carefully defined, computed tomography (CT) cut is greater than those previously reported for systems designed for use with plain radiographs. DESIGN: Observer review of selected cases. SETTING: Four, level one, trauma centers. PATIENTS: Pretreatment CT scans of patients with calcaneus fractures were screened by the authors. Thirty cases were selected that had an appropriate semicoronal CT image. Ten orthopaedic traumatologists who were members of the Orthopaedic Trauma Association and had a minimum of 5 years postresidency experience were selected as reviewers. INTERVENTION: The reviewers were provided with a digital CT image for each case as well as written and diagrammatic representations of the Sanders classification system. The observers then classified each fracture according to the Sanders classification. RESULTS: : The mean kappa value for interobserver reliability for fracture types I-IV was 0.41 +/- 0.02 (mean +/- standard error of the mean; range, 0.07-0.64). Observers disagreed by more than 1 fracture type (ie, I vs. III or II vs. IV) in 10% of the cases. Observers agreed on the location of the fracture lines (A, B, C) in 90% of type II fractures and 52% of type III fractures. CONCLUSIONS: The results indicate that in a carefully controlled paradigm, the interobserver reliability with a classification system based on interpretation of a single, carefully defined CT image was no better than the results reported for the same classification system used with full CT data or for other classification systems used for various fractures in the skeleton. Agreement in identifying the location of the fracture lines was very good for simple fractures but much worse for complex injuries. Additional study may determine whether the use of a full complement of CT images can improve reliability in classification of complex injuries. PMID- 16247307 TI - Posttraumatic stress symptoms in children recovering from minor orthopaedic injury and treatment. AB - BACKGROUND: This study was designed to determine the prevalence of high levels of posttraumatic stress disorder (PTSD) symptoms among pediatric orthopaedic trauma patients recovering from injury and to see whether injury or demographic variables are associated with the presence of the symptoms. METHODS: Four hundred pediatric orthopaedic trauma patients completed the Child PTSD Symptom Scale questionnaire. Demographic and injury variables were tested to see if any were associated with the presence of high levels of posttraumatic stress symptoms. RESULTS: The average age of respondents was 11 years. The average time since injury was 36 days. The mean Injury Severity Score and summed Extremity Abbreviated Injury Score were 4 and 2, respectively. A total of 130 (33%) met criteria for high levels of PTSD symptoms. None of the variables tested were associated with high levels of PTSD symptoms, except one. Patients admitted to the hospital after injury were significantly more likely to develop high levels of PTSD symptoms. CONCLUSIONS: High levels of posttraumatic stress disorder symptoms are common in the recovery period after pediatric orthopaedic trauma, even among patients with relatively minor injury. Children admitted to the hospital after injury are at higher risk for such symptoms. PMID- 16247308 TI - BMP-14 deficiency inhibits long bone fracture healing: a biochemical, histologic, and radiographic assessment. AB - OBJECTIVES: Bone morphogenetic proteins (BMPs) represent a distinct subset of the transforming growth factor-beta family best known for their role in joint formation and bone growth, and several recent clinical trials have begun to look at their efficacy in the augmentation of fracture healing. The goal of this research is to examine the effect of BMP-14, also known as growth differentiation factor-5 and cartilage- derived morphogenetic protein-1 (GDF-5, CDMP-1) on fracture healing by studying the long bone repair process in mice with a deficiency in this signaling peptide. METHODS: The animal model used for these studies was the BMP-14 (-/-) brachypodism (bp) mouse. Phenotypically normal heterozygous (+/-) littermates were used as controls. Closed mid shaft femur fractures were created and stabilized with intramedullary fixation in 8-week-old female mice. Forty-eight mice per genotype group were examined. On postoperative days 4, 7, 10, 14, 21, 28, 35, and 42, the mice were killed and the femurs and repair tissue were harvested for analysis. At each time point, the fracture sites were analyzed radiographically, histologically, and biochemically. For all quantitative analyses, the data were normalized and analyzed statistically using a 2-factor ANOVA test. RESULTS: Biochemically, peak values of normalized proteoglycan content were approximately 3 times less in the mutant fractures early in the time course of healing compared with the controls (P < 0.05). Histologically, BMP-14-deficient fractures exhibited a delay in peak area cell density, callus organization, and bone formation compared with controls. Radiographic analysis demonstrated that the peak callus was 2 weeks delayed and approximately 2 times less in the mutants compared with controls (P < 0.05). Radiographic grading of callus also demonstrated a significant difference after day 14. CONCLUSIONS: Based on histologic, radiographic, and biochemical analysis, BMP-14-deficient mice display a short-term delay in healing of approximately 1 to 2 weeks. The observed abnormalities seem to be the result of a delay in cellular recruitment and chondrocyte differentiation in the early stages fracture repair in the absence of BMP-14. These results support the hypothesis that BMP-14 deficiency leads to a delay in fracture healing. Further studies are warranted to more closely examine the role of BMP-14 in normal fracture healing and the mechanism by which it works. PMID- 16247309 TI - Ankle fractures in the elderly: what you get depends on where you live and who you see. AB - OBJECTIVES: This study was performed to determine 1) the rate of ankle fractures in the elderly in the United States stratified by hospital referral region, and 2) whether the percentage of ankle fractures treated surgically is affected by factors, such as fracture location, hospital referral region, concentration of orthopaedists, presence of a teaching hospital in that region, patient age, race, gender, or the number and type of specific medical comorbidities. DESIGN: A 20% sample of Medicare Part B claims from the years 1998 to 2000 was analyzed. PATIENTS/INTERVENTION: The CPT codes for operative and nonoperative treatment of isolated medial malleolar, isolated lateral malleolar, bimalleolar, and trimalleolar fractures were identified. These codes were used to determine the overall rate of ankle fractures and individual fracture types. MAIN OUTCOME MEASUREMENT: : The rate of ankle fractures was evaluated by hospital referral region, patient age (groups of 5 years, aged 65 years or older), gender, and race. The percentage of surgical treatment was determined for each fracture type as the number of surgically treated fractures over the total number of ankle fractures within each subtype and analyzed by fracture type, hospital referral region, and concentration of orthopaedists in that region, presence of a teaching hospital within the hospital service area, patient age, gender, race, and number and type of specific medical comorbidities. Regression was performed by using the above variables. RESULTS: We identified 33,704 ankle fractures: 7.6% were isolated medial malleolar, 50.8% were isolated lateral malleolar, 27.4% were bimalleolar, and 14.2% were trimalleolar fractures. The overall United States average was 4.2 ankle fractures per 1000 Medicare enrollees. The rate of ankle fractures varied by a factor of 8, from 1 per 1000 Medicare enrollees in San Francisco, CA, to 8.3 in Hickory, NC. The rate of ankle fractures was highest in white women at 5.8 and lowest in nonwhite men at 1.5 per 1000 Medicare enrollees. The overall rate of ankle fractures that underwent surgical stabilization was 33%, ranging from 14% in Binghampton, NY, to 72% in Napa, CA. The rate of surgical intervention was 22% for isolated medial malleolar fractures, 11% for isolated lateral malleolar fractures, 58% for bimalleolar fractures, and 74% for trimalleolar fractures. In regression analysis, the factors associated with nonoperative care after ankle fracture were: older age, female gender, increasing number of comorbidities as measured by the Charlson index, presence of diabetes or peripheral vascular disease, and living in a hospital service area that had a designated teaching hospital. Beneficiaries living in areas in which a hospital was a member of the Council of Teaching Hospitals were less likely to receive surgical treatment of their ankle fracture. Increasingly older age was strongly associated with decreased likelihood of having surgical intervention, with each 5 year age grouping progressively less likely to have surgical treatment. The concentration of orthopaedists in the region was not associated with the likelihood of having surgical treatment. CONCLUSIONS: The term ankle fracture involves a wide spectrum of injuries. We found a large variation through the United States in both the rate of ankle fractures and the percentage of those that undergo surgical intervention. PMID- 16247310 TI - Initial radiographic evaluation of the spine after trauma: when, what, where, and how to image the acutely traumatized spine. AB - BACKGROUND: Radiographic evaluation of the spine after an acute traumatic event is an integral part of the initial assessment of the patient. Despite this, an imaging protocol that includes detailed recommendations of the views, modalities, and techniques of radiographic work-up is lacking in the peer-reviewed literature. STUDY PURPOSE: The goal of the current review was to suggest a uniform protocol for initial imaging in the traumatized patient and provide a rationale and guidelines to assist in deciding which studies are indicated and at what time they are best obtained. METHODS: The authors' reviewed the available modern, published, English literature, including both peer-reviewed articles and commonly used textbooks, for recommendations concerning which imaging studies are most effective and cost-efficient in detecting spinal injuries in the acute trauma setting. A list of radiologic studies was compiled. A panel of spine surgeon members of the Spine Trauma Study Group, all of whom were highly experienced in trauma management, evaluated the utility and necessity of these imaging studies. CONCLUSIONS: Surgeons agreed that the mainstay of initial radiographic evaluation of the spine after acute trauma remains plain radiographs. CT scanning remains the best mode of delineating the bony details of a spinal injury and should be used to characterize all bony injuries identified on plain radiographs. As CT technology continues to improve, it is likely to play an even greater role as an initial screening tool. The role for MRI continues to expand, particularly in detecting the soft tissue components of injuries. PMID- 16247311 TI - The syringe technique for minimally invasive bone grafting. AB - Standard techniques of bone grafting for nonunion involve extensive dissection of the soft tissues surrounding the nonunion site to gain exposure and facilitate placement of the graft. Successful minimally invasive grafting has been described; however, it can involve difficulties in graft placement or require the use of advanced, expensive technical equipment. We describe a simple, inexpensive technique in which the distal end of a standard syringe is trimmed to provide a conduit for minimally invasive delivery of bone graft material. Three illustrative case reports are discussed in which this technique achieved successful union without operative morbidity. PMID- 16247312 TI - Penetration of the distal femoral anterior cortex during intramedullary nailing for subtrochanteric fractures: a report of three cases. AB - Three cases of anterior distal femoral cortex penetration during intramedullary nailing for subtrochanteric fractures are documented. Case 1 involved a Zimmer (Warsaw, IN) M/DN antegrade femoral nail, case 2 a Howmedica (Allendale, NJ) long stem Gamma nail, and case 3 a Synthes (Paoli, PA) titanium femoral nail with spiral blade locking. The anterior Zimmer nail penetration resulted in a displaced supracondylar fracture, which subsequently required revision. The Gamma nail as well as the Synthes nail were left impaled through the distal femoral cortex, and the subtrochanteric fractures went on to union. The anteroposterior radius of curvature for the Zimmer, the long Gamma, and the Synthes nail are 257 cm, 300 cm, and 150 cm, respectively. It has been estimated that the radius of curvature of the femoral diaphyseal canal is 114 to 120 cm. It appears that the difference in femoral anteroposterior bow between the bone and the implant is a contributing factor to distal femoral anterior cortex penetration in intramedullary nailing of subtrochanteric fractures. PMID- 16247313 TI - Skin breakdown following circumferential pelvic antishock sheeting: a case report. AB - Hemodynamic instability in the setting of high-energy blunt trauma is a challenging clinical scenario. Rapid provisional stabilization of the unstable pelvis is advisable because it reduces ongoing blood loss and pain by imparting bone and soft tissue stability as well as reducing pelvic volume. The use of a standard bed sheet has become a popular choice for achieving temporary stability of the pelvis through a technique that has been called circumferential pelvic antishock sheeting. Although we have found circumferential pelvic antishock sheeting to be a valuable tool in our institution, we describe a case of skin compromise that precluded complete internal fixation of a complex pelvic ring/acetabular fracture. PMID- 16247314 TI - Permanent articulated antibiotic-impregnated cement spacer in septic shoulder arthroplasty: a case report. AB - We report a case of a patient with an infected shoulder hemiarthroplasty in whom a permanent antibiotic-impregnated cement spacer was employed with satisfactory results. This method of treatment has limited applications and would not be appropriate in all cases of septic shoulder joint arthroplasties. However, its use may represent a valid alternative in low physical demand patients who are unwilling to undergo major surgery or when inadequate bone stock is present. PMID- 16247315 TI - Progressive cubitus varus due to a bony physeal bar in a 4-year-old girl following a supracondylar fracture: a case report. AB - We report a case of progressive cubitus varus deformity caused by a physeal bar following a supracondylar humeral fracture in a 4-year-old girl. Malreduction is thought to be the commonest cause of this deformity, which is nonprogressive. A corrective osteotomy in cases like ours should be deferred until skeletal maturity. PMID- 16247316 TI - Position: operative treatment. PMID- 16247317 TI - Position: nonoperative management. PMID- 16247324 TI - Hepatosplanchnic circulatory dysfunction in acute hepatic infection: the case of dengue hemorrhagic fever. AB - The mechanism of shock in patients with dengue hemorrhagic fever (DHF) has not yet been fully understood. In this study, we investigated the possibility of splanchnic venous pooling as a contributor for circulatory dysfunction in these patients. Ultrasonographic studies of portal vein and inferior vena cava were done in 45 patients with serologically or PCR-confirmed diagnosis of dengue virus infection. The size of portal vein and inferior vena cava, mean blood flow velocity in the right portal vein, and modified portal vein congestion index were compared between patients with dengue fever (DF, n = 20), DHF without shock (n = 14), and dengue shock syndrome (DSS, n = 11) during the toxic stage, convalescent stage, and at follow-up. The portal vein was significantly more dilated in patients with shock (DSS) than DHF without shock and than DF during the toxic and convalescent stages (P < 0.05), but not at follow-up. The change in the size of inferior vena cava followed the opposite trend (not statistically significant). Portal vein blood flow velocity was lower and congestion index was higher in shock cases (DSS) than DHF without shock and than DF at toxic and convalescent stages (P < 0.01). The differences disappeared at follow-up. Hepatosplanchnic venous pooling and/or dysfunction occur and correlate with the severity of circulatory derangement and shock in patients with DHF. The cause(s) and significance of hepatosplanchnic circulatory dysfunction in DHF and possibly other viral hepatic diseases deserve further study. PMID- 16247325 TI - The severity of cholestatic injury is modulated by the genetic background. AB - Common bile duct ligation (CBDL) compromises the hepatic reticuloendothelial system by impairing the clearing of endotoxin and triggering an overwhelming inflammatory response. The response to endotoxin at the level of cytokine release and subsequent mortality depends on the genetic background in experimental mouse models. We hypothesized that the genetic make-up modulates the inflammatory responses after CBDL. The CBD was ligated in male A/J and B6 mice (8 weeks old). At 7 days post-CBDL, the presence of ascites was observed in 80% of B6 mice but in none of the A/J mice (P < 0.001). B6 mice showed higher mortality than A/J mice (P < 0.05). Both strains had marked cholestatic injury documented histologically. Liver chemistries were markedly elevated in both strains after injury. Plasma levels of the anti-inflammatory cytokine IL-10 were significantly higher in A/J than B6 mice at the 4- and 12-h time points (P < 0.05), whereas proinflammatory cytokine TNF-alpha levels were significantly higher in B6 than A/J mice at 2 h (P < 0.05). Both strains displayed activation of NF-kappaB after CBDL. In conclusion, the contrasting response observed after CBDL between A/J and B6 mice is largely attributable to genetic differences. Survival after CBDL was correlated with an increase in anti-inflammatory cytokines. PMID- 16247326 TI - Zaprinast attenuates hypoxic pulmonary artery injury and causes less aortic relaxation than milrinone. AB - Hypoxic pulmonary vasoconstriction is a challenging clinical problem with limited therapeutic options. Milrinone, a phosphodiesterase (PDE)-3 inhibitor, is frequently used to treat perioperative pulmonary hypertension. However, recent evidence suggests that the PDE-5 isoform may be more specific for lung tissue. We hypothesized that the PDE-5 inhibitor zaprinast has greater efficacy for pulmonary vasorelaxation, attenuation of hypoxic pulmonary vasoconstriction, and inhibition of hypoxia-induced pulmonary artery cytokine expression when compared with milrinone. To study this, isolated rat pulmonary artery and thoracic aorta rings suspended in physiologic organ baths for measurement of isometric force transduction were treated with vehicle (dimethyl sulfoxide), milrinone, or zaprinast to assess pulmonary artery relaxation, thoracic aorta relaxation, inhibition of hypoxic (pO2 = 30-35 mmHg) pulmonary vasoconstriction, and hypoxia induced pulmonary artery TNF-alpha and IL-1beta expression (reverse transcriptase PCR). Milrinone and zaprinast resulted in dose-dependent pulmonary artery and aortic relaxation, but zaprinast caused significantly less aortic relaxation compared with milrinone (50.12% +/- 3.36% versus 91.03% +/- 2.97%, P < 0.001). Zaprinast, but not milrinone, significantly inhibited hypoxic pulmonary vasoconstriction (zaprinast, 58.42% +/- 5.37%; milrinone, 77.65% +/- 4.42% versus vehicle: 74.42% +/- 7.54%). Hypoxia-induced upregulation of TNF-alpha and IL 1beta mRNA in pulmonary artery was decreased by zaprinast, but not milrinone, pretreatment. These results suggest that zaprinast, but not milrinone, preferentially vasodilates pulmonary artery over aorta, attenuates hypoxic pulmonary vasoconstriction, and inhibits hypoxia-induced pulmonary artery TNF alpha and IL-1beta expression. Therefore, PDE-5 inhibition may be advantageous in the treatment of pulmonary hypertension. PMID- 16247327 TI - Age-dependent responses to hepatic ischemia/reperfusion injury. AB - The current study explored the concept that adult and pediatric populations differ in their response to major injury. Male C57BL/6 mice of a "young adult" (8 12 weeks) or "mature adult" (12-13 months) age were subjected to partial hepatic ischemia and reperfusion. Mature adult mice displayed significantly more liver injury than young adult mice as assessed histologically and by serum levels of alanine aminotransferase. Interestingly, there was far less neutrophil accumulation in the livers of mature adult mice. However, liver-recruited neutrophils from mature adult mice had a higher activation state than those from young adult mice. Activation of the inflammatory transcription factor, NF-kappaB, was suppressed in whole livers from mature adult mice. In isolated liver cells, Kupffer cells showed no difference in NF-kappaB activation, but hepatocytes from mature adult mice had delayed NF-kappaB activation in response to TNF. Furthermore, isolated hepatocytes from young adult mice produced abundant amounts of the chemokine, macrophage inflammatory protein-2, whereas hepatocytes from mature adult mice produced little, if any macrophage inflammatory protein-2. Mature adult mice had much lower hepatic expression of the cytoprotective protein, heat shock protein 70, than did young adult mice. In contrast, serum heat shock protein 70 levels, which has been linked to subsequent tissue injury, were higher in mature adult mice than in young adult mice. These data suggest that there are multiple alterations at the cellular and molecular levels that contribute to enhanced postischemic liver injury in mature adult mice. PMID- 16247328 TI - Blood-brain barrier changes during compensated and decompensated hemorrhagic shock. AB - Dysfunction of the blood-brain barrier (BBB) can be associated with a large number of central nervous system and systemic disorders. The aim of the present study was to determine BBB changes during different phases of hemorrhagic shock. The experiments were carried out on male Wistar rats anaesthetized with urethane. To produce compensated or decompensated hemorrhagic shock, mean arterial pressure was decreased from the normotensive control values to 40 mmHg by a standardized method of blood withdrawal from the femoral artery. Cerebral blood flow changes were followed by laser-Doppler flowmetry, and arterial blood gas values were monitored over the whole procedure. Cortical blood flow was significantly reduced in compensated and in decompensated hemorrhagic shock compared with the normotensive rats. As the shock shifted to the decompensated phase, the blood flow reduction was more pronounced. BBB permeability studies using sodium fluorescein (molecular weight of 376) and Evan's Blue albumin (molecular weight of 67,000) have revealed a significant increase of the BBB permeability for sodium fluorescein in the decompensated stage of hemorrhagic shock. Western blot analysis of brain capillaries showed that the expression of the transmembrane tight junction protein occludin was reduced in response to hemorrhagic shock, and the decrease of occludin was more pronounced in the decompensated phase. A similar expression pattern was shown by the transmembrane adherens junction protein cadherin as well. Our results suggest that the decompensated phase of hemorrhagic shock is associated with disturbances of the BBB, which may be explained by the dysfunction of interendothelial junctions caused by decreased occludin and cadherin levels. PMID- 16247329 TI - Hydroxyethyl starch inhibits neutrophil adhesion and transendothelial migration. AB - A resuscitation strategy that significantly alters the state of neutrophil (PMN) activation may impact organ function and survivability after shock. Various resuscitative fluids have been shown to elicit a severe immune activation and an upregulation of cellular injury markers, whereas other fluids have been shown to be protective. Recent studies have demonstrated that hydroxyethyl starch (HES), an artificial colloid, may exert significant anti-inflammatory effects, whereas conflicting studies with the same substance have shown an increase in PMN activation. Successful manipulation of the early immune events associated with hemorrhage and resuscitation will require a better understanding of the possible pro- or anti-inflammatory effects of resuscitation fluids. Our study investigated the effect of HES directly on PMN and cultured vascular endothelial cells in vitro. The effect of HES on PMN surface expression of CD11b and L-selectin was measured by flow cytometry. PMN activation response to HES was measured using a shape-change assay in response to formyl-methionyl-leucyl-phenylalanine (f-MLP). The effect of HES on endothelial cell surface expression of E-selectin, P selectin, vascular cell adhesion molecule-1(VCAM-1), and intracellular adhesion molecule-1 (ICAM-1) was evaluated by enzyme-linked immunoabsorbant assay. PMN rolling, adhesion, and migration events were measured using direct microscopy under conditions simulating microvascular flow. PMN surface expression of CD11b and L-selectin in whole blood samples and isolated PMNs were unaffected by exposure to HES. HES had no effect on the normal f-MLP dose-dependent increase in PMN activation. In the absence of IL-1 stimulation, there was a small but statistically significant (P < 0.05) increase in ICAM-1 after exposure to HES. After stimulation with IL-1 (10 U/mL), HES had no effect on the expression of P selectin, E-selectin, ICAM-1, or VCAM-1. Under simulated microvascular flow conditions in vitro, HES significantly diminished the PMN tethering rate (P < 0.05) and the transendothelial migration rate (P < 0.05) in a dose-dependent manner. HES significantly alters the function of the PMN at the interface of the PMN responding to activated endothelium. The effect occurs, surprisingly, without a coincident effect on the state of PMN activation or a significant change in the surface expression of the adhesion molecules responsible for PMN-endothelial interaction. PMID- 16247330 TI - ATP-induced calcium increase as a potential first signal in mechanical tissue trauma. A laser scanning microscopic study on cultured mouse skeletal myocytes. AB - Although it is known that after major tissue trauma, local incidents in the mechanically destroyed muscle tissue form the basis of subsequently occurring severe inflammatory reactions, the very first events taking place immediately after myocyte destruction have not been studied on the single cell level thus far. Therefore, in this study, the reaction of cultured C2C12 mouse skeletal myocytes to lethal injury was examined using laser scanning microscopy. Mechanical rupture of one single myocyte induced an immediate accumulation of calcium in its cytosol and nuclei, as detected by an increase in the fluorescence intensity of the intracellular calcium-sensitive dye Fluo-3. The intracellular calcium elevation propagated further to the adjacent, noninjured myocytes in a wave-like fashion within seconds. The calcium increase detected in these neighboring cells was higher and up to 1000 times more extended than the physiological calcium spike that induces C2C12 myocyte contraction. Wave propagation did not depend on gap junctional communication but occurred via liberation of nucleotides, mainly ATP, but presumably also UTP and others, from the destroyed cell and subsequent calcium release from the sarcoplasmic reticulum via a purinoceptor-mediated mechanism in the adjacent cells. These findings suggest a decisive role of ATP and related nucleotides in the pathogenesis of tissue trauma because they appear to initiate the signaling mechanism from injured myocytes to the surrounding tissue and potentially to the whole body. PMID- 16247331 TI - Chronic ethanol sensitizes the liver to endotoxin via effects on endothelial nitric oxide synthase regulation. AB - In vivo studies have shown that chronic alcohol consumption sensitizes the liver to endotoxemic shock, leading to liver microcirculation disruption. In the present study, we investigated the molecular mechanisms involved, focusing on endothelial nitric oxide synthase (eNOS) activity and regulation, which represents one of the major vasodilatory pathways. Male Sprague-Dawley rats were fed an alcohol liquid diet or a control isocaloric diet for 5 weeks. Priming effects of ethanol were studied in a model with or without a 24-h LPS treatment (1 mg/kg body weight). At the end of the diet, liver tissue was harvested for western blot, reverse transcriptase-PCR, histological analysis, and immunostaining and blood for serum alanine aminotransferase analysis. Chronic ethanol and LPS alone induced a mild hepatitis and infiltration, respectively. Combined, LPS and chronic ethanol feeding showed a synergistic effect on the liver, leading to extensive steatohepatitis with extensive focal necrosis associated with significantly higher levels of serum ALT. Chronic ethanol and LPS significantly inhibited eNOS activity, but exerted their effects through different mechanisms. Caveolin-1, an eNOS inhibitory protein, was upregulated after LPS and chronic alcohol consumption. Additionally, chronic alcohol consumption down-regulated endothelin B receptor, eNOS protein levels, and eNOS phosphorylation. In conclusion, chronic ethanol consumption and LPS share a similar pathophysiology and both lead to the impairment of eNOS activity, but through distinct molecular mechanisms. The presence of focal necrosis in a mild stress model could provide a good animal study to investigate the advanced stages of alcoholic liver diseases. PMID- 16247332 TI - Overexpression of alphaB crystallin in the gastrointestinal tract of the newborn piglet after hypoxia. AB - Newborn animals are particularly sensitive to hypoxic stress. Oxygen is spared for sensitive tissues, including brain and heart. Scarce information is available concerning the molecular effects of hypoxia in the gastrointestinal tract (GIT). Moreover, stress protein expressions, including heat shock proteins (HSP), are still poorly documented in the GIT. Our objective was to determine the possible effect of hypoxia on HSP expression at birth. After western blotting, alphaB crystallin, HSP 27, and HSP 70 expressions were determined in newborn controls and piglets exposed to 1 or 4 h hypoxia (5% O2, 95% N2) allowed to recover from 1 to 68 h. Cytosol and nuclei were also separated and the extracts were tested for HSF1 and alphaB crystallin expressions. Surprisingly, alphaB crystallin was overexpressed in the stomach and colon in animals submitted to hypoxia, whereas HSP 27 and HSP 70 expression remained stable. Increases and return to basal levels in HSF1 and alphaB crystallin were simultaneously observed in the unique nuclear compartment. To our knowledge, the present study is the first to demonstrate the oxygen dependency of an HSP in the GIT, particularly in the colon in newborn piglets. The kinetics of alphaB crystallin overexpression after hypoxia parallels the activation of HSF1. This observation possibly indicates a correlation between this factor and alphaB crystallin after hypoxia. Taken together, the present results open the field of wide investigation about the specific response of this low-molecular-weight HSP and its possible involvement in pathological states in the GIT such as stomach and colon. PMID- 16247333 TI - Peroxisome proliferator-activated receptor gamma mediates protection against cyclooxygenase-2-induced gut dysfunction in a rodent model of mesenteric ischemia/reperfusion. AB - Cyclooxygenase (COX)-2 has been identified as an important mediator elaborated during ischemia/reperfusion, with pro- and anti-inflammatory properties having been reported. As the role of COX-2 in the small intestine remains unclear, we hypothesized that COX-2 expression would mediate mesenteric ischemia/reperfusion induced gut injury, inflammation, and impaired transit and that these deleterious effects could be reversed by the selective COX-2 inhibitor, N-[2-(cyclohexyloxy) 4-nitrophenyl] methanesulphanamide (NS-398). Additionally, we sought to determine the role of peroxisome proliferator-activated receptor gamma (PPARgamma) in mediating protection by NS-398 in this model. Rats underwent sham surgery or were pretreated with NS-398 (3, 10, or 30 mg/kg) intraperitoneally 1 h before 60 min of superior mesenteric artery occlusion and 30 min to 6 h of reperfusion. In some experiments, NS-398 (30 mg/kg) was administered postischemia. Ileum was harvested for COX-2 mRNA and protein, PGE2, myeloperoxidase (inflammation), histology (injury), intestinal transit and PPARgamma protein expression, and DNA-binding activity. COX-2 expression and PGE2 production increased after mesenteric ischemia/reperfusion and were associated with gut inflammation, injury, and impaired transit. Inhibition of COX-2 by NS-398 (30 mg/kg, but not 3 or 10 mg/kg) not only reversed the deleterious effects of COX-2, but additionally induced expression and nuclear translocation of PPARgamma. NS-398 given postischemia was equally protective. In conclusion, COX-2 may function as a proinflammatory mediator in a rodent model of mesenteric ischemia/reperfusion. Reversal of gut inflammation, injury, and impaired transit by high-dose NS-398 is associated with PPAR activation, suggesting a potential role for PPAR-gamma in shock-induced gut protection. PMID- 16247334 TI - Subclass opioid receptors associated with the cardiovascular depression after traumatic shock and the antishock effects of its specific receptor antagonists. AB - The present available opioid receptor antagonists such as naloxone and naltrexone are not highly receptor selective. They may antagonize mu opioid receptors to affect the pain threshold of the patients with traumatic shock while they exert antishock effects. Therefore, they are not suitable for traumatic shock. It is very important to elucidate the subclass of opioid receptors that are closely associated with cardiovascular depression of traumatic shock and then choose their specific receptor antagonists to treat it. Traumatic shock was used in pentobarbital-anesthetized Wistar rats by right femur fracture plus hemorrhage (fixed hemorrhage at a rate of 20 mL/kg in experiment 1 or hemorrhage to 40 mmHg mean arterial blood pressure for 60 min in experiments 2 and 3), and the changes of myocardial and brain opioid receptors after traumatic shock, the antagonizing effects of mu, delta, and kappa opioid receptor antagonists on the cardiovascular depression of traumatic shock and the antishock effects of delta and kappa opioid receptor antagonists ICI174,864 and Nor-binaltorphimine (Nor-BNI) were observed. The results indicate that after traumatic shock, the number of myocardial and brain delta and kappa opioid receptors were significantly increased that were significantly associated with the decreased cardiovascular functions. mu Opioid receptors in the heart and brain did not change significantly. Intracerebral ventricular administration of ICI174,864 and Nor-BNI significantly antagonized the decreased cardiovascular function after traumatic shock and increased the survival rate of traumatic shock rats, but mu opioid receptor antagonist beta funaltrexamine did not. Meanwhile, intravenous administration of delta and kappa opioid receptor antagonists ICI174,864 and Nor-BNI also significantly increased the mean arterial blood pressure, improved the hemodynamic parameters, and prolonged the survival rate of traumatic shock rats. These findings suggest that opioid receptors are involved in the cardiovascular depression of traumatic shock, and the subclass receptors are mainly delta and kappa opioid receptors. delta and kappa opioid receptor antagonists have good beneficial effects on traumatic shock. PMID- 16247335 TI - Effects of intrarenal administration of the cox-2 inhibitor parecoxib during porcine suprarenal aortic cross-clamping. AB - The aim of this study was to investigate the effects of intrarenal administration of the cyclooxygenase-2 inhibitor parecoxib during suprarenal aortic cross clamping. In a prospective, controlled, blinded, randomized manner, 16 anesthetized and mechanically ventilated pigs were instrumented to measure systemic and right kidney hemodynamics, oxygen exchange, and metabolism. During 45 min of suprarenal aortic cross-clamping, animals received 40 mg of parecoxib (n = 8) or vehicle (n = 8) infused continuously into the right renal artery. Hemodynamic and metabolic data, right kidney venous blood, as well as urine samples were obtained before clamping, as well as before and 75 and 195 min after declamping. Clamping transiently increased mean arterial pressure in both groups. Systemic and renal blood flow did not differ between the pre- and postclamping measurements or between groups. Parecoxib attenuated the otherwise significant fall in right kidney creatinine clearance (controls: from 45 [7;111] to 17 [9;22] mL/min; parecoxib: from 39 [3;59] to 27 [11;45] mL/min, P = 0.039 and P = 0.297, respectively versus before clamping, P = 0.021 versus controls at 195 min) and prevented the impairment of renal lactate balance observed in the control group (controls: from 0.5 [-0.8;3.5] to 0.2 [-0.2;0.6] mumol/kg/min; parecoxib: from 0.6 [-1.0;2.0] to 0.4 [-1.2;0.6] mumol/kg/min, P = 0.038 and P = 0.285, respectively, versus before clamping). In conclusion, intrarenal parecoxib infusion beneficially influenced kidney function in this clinically relevant model of suprarenal aortic cross-clamping. PMID- 16247336 TI - Oral administration of geranylgeranylacetone improves survival rate in a rat endotoxin shock model: administration timing and heat shock protein 70 induction. AB - The present study was performed to determine whether oral pretreatment with geranylgeranylacetone (GGA) inhibits proinflammatory cytokine liberation and nitric oxide (NO) production in lipopolysaccharide (LPS)-treated rats and protects rats against death from LPS-induced endotoxin shock, and whether such protection by GGA is related to heat shock protein (HSP) 70 induction in multiple organs of rats. GGA (200 mg/kg) was given orally to rats. LPS (20 mg/kg) was administered intraperitoneally 4, 8, 16, or 24 h after GGA administration. The survival of rats was monitored over 24 h after LPS administration. GGA treatment at 8 or 16 h before LPS dramatically improved the survival rate of LPS-treated rats. Plasma levels of proinflammatory cytokines (tumor necrosis factor-alpha and interleukin-6) and NO 6 h after LPS administration in these GGA-pretreated rats were less than one-half of those in rats treated with LPS alone. A GGA challenge 8 or 16 h before LPS administration enhanced HSP70 expression in rat organs after LPS. Treatment with GGA 8 h before LPS minimized hepatic and renal damage. Furthermore, the protective effect of GGA on mortality in LPS-treated rats was inhibited with quercetin, known as an HSP70 inhibitor. These results suggest that oral administration of GGA at an optimal time before LPS injection induces and enhances HSP70 expression in several organs, inhibits proinflammatory cytokine and NO production, and prevents organ damage, resulting in an improved survival rate. PMID- 16247338 TI - Murine in vivo myocardial contractile dysfunction after burn injury is exacerbated by pneumonia sepsis. AB - We evaluated hemodynamic and cardiac contractile dysfunction in a murine model of 40% contact burn complicated by Streptococcus pneumoniae (1 x 10(5) CFU) sepsis. Male, 9- to 10-week-old C57/BL6 mice were divided into the following groups: sham burn, sham sepsis; 24 h after burn alone; 24 h after sepsis alone; 7 days after burn alone; and 7 days after burn followed by pneumonia sepsis. Hemodynamic and cardiac contractile function was assessed with carotid artery cannulation and left ventricular pressure-volume analysis. At 24 h after burn, there were significant decreases in all load-insensitive contractility variables including the end-systolic pressure volume relationship, preload-recruitable stroke work, and maximum elastance, but there were no changes in global hemodynamics. Twenty four hours after sepsis, there was similar cardiac contractile dysfunction, along with a decrease in cardiac output, but mean arterial pressure was maintained with an increase in systemic vascular resistance. Late burn (7 days) was associated with a recovery of all contractility variables except the end-systolic pressure volume relationship. However, sepsis induced during the late burn period was associated with a significant decrease in heart rate and cardiac output, but mean arterial pressure was still maintained with increased systemic vascular resistance. With burn complicated by sepsis, all cardiac contractility variables showed profound contractile dysfunction. Our data indicate that burn complicated by sepsis is associated with more pronounced cardiac contractile dysfunction than burn alone or sepsis alone. PMID- 16247337 TI - Role of p38 mitogen-activated protein kinase on renal dysfunction after hemorrhagic shock in rats. AB - Hemorrhagic shock has been reported to induce renal dysfunction as a consequence of different kinds of local inflammatory response. p38 mitogen-activated protein kinase (MAPK) is a key mediator in organ dysfunction relating to the inflammatory states, and acts as an important mediator in the intracellular signal pathway for proliferation, differentiation, and production of proinflammatory cytokines such as TNF-alpha and IL-1beta. The effect of p38 MAPK on the hemorrhagic damage has not been clearly estimated as yet. In this study, our aim was to evaluate the role of p38 MAPK on the renal damage during the first 5 h after a hemorrhage using a specific inhibitor of p38 MAPK activation, FR167653. p38 MAPK activation increased immediately after a hemorrhage and decreased with time. renal mRNA expression of TNF-alpha and IL-1beta increased, renal dysfunction continued to progress, and histological inflammatory injuries were confirmed after hemorrhagic shock. With the pretreatment of FR167653, all of these hemorrhagic changes were attenuated, although the induction of the primary hypotensive state was confirmed. This study demonstrated that renal p38 MAPK is activated in hemorrhagic shock, promotes the expression of proinflammatory cytokines in the kidney, and consequently develops renal dysfunction. We concluded that p38 MAPK activation is essential in causing renal damage and that the inhibition of p38 MAPK activation blocks the development of the renal dysfunction after hemorrhagic shock. PMID- 16247339 TI - Second generation antipsychotics--aripiprazole revisited. AB - Six second-generation antipsychotics are marketed in the US. Is aripiprazole as effective as the others and better tolerated? PMID- 16247340 TI - InstaRead Lithium System. AB - A finger-stick office test for measuring serum lithium concentrations. PMID- 16247341 TI - Tipranavir (Aptivus) for HIV. AB - A new protease inhibitor, used with ritonavir, for resistant HIV. PMID- 16247342 TI - Pure surgical treatment of 109 aneurysms. AB - AIM: Target of this study was to investigate outcomes after pure surgical treatment of intracranial aneurysms. METHODS: Patients with intracranial supratentorial circle aneurysms were retrospectively reviewed between July 1994 and October 1998. Studied cases were admitted at the Department of Neurosurgery of S. Maria-Hospital, Terni, a Government supported General Hospital. One hundred and nine Hunt and Hess Grade 0 to III patients with supratentorial circle aneurysms was studied in order to determine whether advances in the surgical management of intracranial aneurysms have improved surgical outcomes and which factors may predict outcome. All patients were managed only with standard neurosurgical aneurysms clipping procedures. Outcomes evaluation was made at patients' discharge and classified on the base of the Glasgow Outcome Scale (GOS). Surgical timing, SAH grading, pre and post surgical symptomatic vasospasm, temporary clipping, and intraoperative aneurysm rupture were correlated with outcomes. RESULTS: Surgical results showed a 75% excellent outcome. Mortality rate was 3%. Hunt and Hess grade 0 highly influenced outcome. Differences in outcomes among grades I to III were not significant. No differences in outcomes related to temporary clipping were noted. A low rate of intraoperative aneurysm rupture is reported: 5 out of 109 cases. In all these cases outcome was good, with neither mortality or morbidity. CONCLUSIONS: Results indicate a progressive improvement in surgical outcomes, suggesting that there still exist margins for improvements in pure surgical management of intracranial aneurysms. PMID- 16247343 TI - Chronic subdural hematoma: comparison of two surgical techniques. Preliminary results of a prospective randomized study. AB - AIM: The optimal treatment for chronic subdural hematoma (CSDH) is not yet well defined and research for efficient surgical solutions continues. Burr hole craniotomy (BHC) is a common treatment and twist drill craniostomy (TDC) is a less invasive alternative. A closed-system drainage with subdural expansion catheter and suction reservoir can be used to enhance the TDC procedure. METHODS: We report preliminary results of a prospective study comparing BHC and TDC with suction drainage, in a series of 47 patients randomized into two treatment groups. One group of 22 patients underwent TDC with closed-system drainage and suction reservoir (14 men, 8 women, mean age 78.7 years). Another group of 24 patients underwent BHC with subdural irrigation and closed drainage (16 men, 8 women, mean age 76.3 years). Neurological status was assessed by Markwalder's Grading Scale on admission and at follow-up. All patients underwent computed tomography (CT) before surgery, within 4 days after it, and 1 and 2 months later. RESULTS: Preoperative clinical and radiological data were similar in the two groups. Operating times were shorter in the TDC group, while drainage time was shorter in the BHC group (P<0.0001). Length of hospitalization was similar. Recurrence rate, mortality, and neurological recovery were similar, with non significant differences in favor of the TDC group. After 2 months, CT showed complete regression of subdural effusion in 66.6% of cases in the TDC group and in 31.8% in the BHC group (P<0.05). CONCLUSIONS: Preliminary results indicate TDC and BHC as at least equally effective, however TDC favors faster regression of residual subdural effusion, is a faster procedure, and seems to be associated with fewer recurrences. If confirmed at the end of the study, these results could indicate TDC with suction as the elective surgical treatment for CSDH. PMID- 16247344 TI - Surgical approaches to the cervico-thoracic junction. AB - The cervico-thoracic junction (CTJ) extends between the 7th cervical and the 4th thoracic vertebrae and comprehends the inferior portion of the brachial plexus and the parenchymatous, vascular and nervous structures of the upper mediastinum. The posterior surgical approaches, as the laminectomy or the arthro-pediclectomy, fail to expose the anterior spinal elements. Thus, further surgical approaches have been proposed: postero-lateral, antero-lateral (thoracotomies) and purely anterior. The aim of this study was to discuss indications, key anatomical landmarks and risks of the main surgical approaches to the CTJ. Ten fresh cadavers from the Anatomical Laboratory of the University of Nantes (France) were used for the surgical dissection of the CTJ. The postero-lateral and the antero lateral approaches were performed in 4 cadavers each and the anterior approaches were studied in 2. The postero-lateral extrapleural approach (PLEA) permits an excellent antero-lateral exposure of the T2-T4 segment, preserving the parascapular musculature integrity. The thoracotomies allow the exposure of the antero-lateral portion of the junctional vertebrae, with the limits of the intrapleural approaches. The anterior approaches, including the presternocleidomastoid cervicotomy eventually associated to the sterno claviculotomy, expose the anterior portion of the cervical and the upper thoracic vertebrae up to T4. We believe that the PLEA performs the greater surgical exposure with minimal risk of vasculo-nervous damage. Among the anterior approaches, the simple cervicotomy is the most indicated procedure in case of patients with certain anatomical conditions. PMID- 16247345 TI - "Shave" biopsy of the optic nerve in isolated neurosarcoidosis. AB - We report a case of isolated central nervous system sarcoidosis, limited to the optic nerve, characterized by negative systemic work up and neuroradiological features consistent with both optic sheath meningioma and optic nerve glioma. A magnetic resonance imaging examination revealed a dural tail sign associated with a diffuse enhancement and enlargement of the optic nerve. Both positive laboratory findings and systemic manifestation of sarcoidosis or autoimmune diseases were absent. Because of progressive visual loss, tissue confirmation was sought and the optic nerve itself biopsied after non-diagnostic dural samplings. The approach to this was to perform a "shave" biopsy of the optic nerve within its intracranial compartment. This procedure spared the central macular fibers and was not associated with loss of vision. The use of the fiber-sparing "shave" biopsy provided diagnostic samples containing non-caseating granulomas, without compromising the patient's preoperative central visual field. The authors advocate the use of this simple and safe technique when a definitive diagnosis among optic nerve meningioma, optic nerve glioma and isolated neurosarcoidosis cannot be reached by means of less invasive procedures. PMID- 16247346 TI - [Overactive bladder: modulating the effects of antimuscarinic therapy]. AB - Overactive bladder (OAB) is a prevalent pathologic condition affecting millions of young, adult and aging people in the world. Although it is a underestimated disease, it causes a significant negative socio-economic impact and determines a severe deterioration of quality of life (QoL) of sufferers. The epidemiologic data today available regarding OAB incidence, prevalence and burden, have prompted a great interest about it. The effect of this research resulted in the availability of new clinical tools useful to investigate and easier diagnose OAB, such as voiding diary and specific symptomatic questionnaires. The most used drugs in the OAB treatment are the antimuscarinics, whose use is still limited due to their some side effects, mostly dry mouth and constipation. For this reason the pharmacologic research is involved in the development of drugs with a better receptorial selectivity, organ specificity, new extended release formulations or new ways of administration. The new extended release formulation of tolterodine, one of the most used drugs in OAB treatment, present a better tolerability maintaining its efficacy, with a consequent better therapeutic efficacy. Furthermore, the recent proposed night-time administration of the drug resulted in a even more lowering of side-effects due to their occurrence during the nocturnal resting. PMID- 16247347 TI - PPAR-gamma-agonists' renal effects. AB - PPAR-gamma ligands, including thiazolidinediones, have recently become clinically available for treating insulin-resistant diabetes mellitus. Accumulating evidence suggests that these drugs not only significantly improve insulin sensitivity but also may have antiproteinuric effects in genetically obese diabetic rodents and patients with type II diabetes and diabetic nephropathy. Moreover, troglitazone reduced expression of ECM proteins and transforming growth factor-beta in glomeruli from streptozotocin-induced diabetic rats. Many other properties including antiproteinuric, hemodynamic, and antihypertensive effects in insulin dependent diabetes mellitus suggest that PPAR-gamma ligands might have a direct, beneficial renal effect, independent of their capacity to improve glucose tolerance. Besides their antidiabetic effects, thiazolidinediones have been shown to lower blood pressure in diabetic patients with hypertension and patients with diabetic nephropathy through multiple mechanisms. Several studies showed the efficacy of PPAR-gamma agonists to ameliorate the progression of glomerulosclerosis. The effect is independent of insulin effects and could only be partially due to lipid effects. These renal protective effects of PPAR-gamma agonists suggest that they may provide a novel intervention strategy to prevent vascular and glomerular sclerosis. PMID- 16247348 TI - Renal tumor radiofrequency ablation. AB - Nephron sparing surgery has become the gold standard for the treatment of small (< 4 cm) renal tumors. More recently, renal tumor ablation, destroying renal tumors with energy based modalities and leaving the tissue in situ, has gained popularity. Radiofrequency ablation (RFA) utilizes heat energy to cause cellular destruction. RFA can be applied laparoscopically, percutaneously, and as an adjunct with partial nephrectomy. A comprehensive MEDLINE search of the English literature from 1990 to present was performed to identify articles on renal RFA. RFA is an effective minimally invasive treatment alternative for small renal masses in patients with significant comorbidities that preclude extirpative surgery. Limited follow up from a number of series demonstrate a success rate of 92%. RFA has proven to be an effective ablative therapy in a number of organ systems. Over the past decade it has been applied to the treatment of small renal tumors. The short-term efficacy and minimum morbidity of RF thermal therapy for renal tumors has been encouraging. RFA provides a versatile tool to add to the urologist's armamentarium in the minimally invasive treatment of renal cancer. PMID- 16247350 TI - Prostate cancer genetics. AB - Prostate cancer (PC) is the most frequent malignant tumor among men over 50 years old. Its incidence varies according to countries and ethnic groups. Known risk factors are race and positive family history of the disease. Familial aggregation (at least 2 cases in the family) is observed in about 20% of cases and a hereditary form of PC, compatible with a Mendelian inheritance, in 5%. This proportion increases with younger age at diagnosis. Eight putative loci are already identified for hereditary PC but undoubtedly, others will be found in forthcoming studies. The genetic heterogeneity observed in hereditary PC reflects variety of origins of the studied families. Agreggation of PC and other cancers in some families suggests the involvement of common susceptibility genes. In other familial form and in sporadic cases, the genetic component should be polygenic: PC wouldn't result from segregation of a major gene mutation transmitted according to a monogenic inheritance, but rather to sharing of alleles at many loci, each contributing to a small increase in cancer risk. Indeed, several genetic polymorphisms were associated with an increased risk of developing PC and could explain the variations of PC incidence observed between populations. PMID- 16247349 TI - Strategies for improved shock wave lithotripsy. AB - Research in lithotripsy that started with the effort to characterize acute shock wave damage to the kidney has led to advances on several fronts, including discovery of strategies that have improved clinical treatment. It is appreciated now that shock wave trauma is primarily a vascular lesion, that injury is dose dependent, and that hemorrhage can be severe and can lead to a permanent loss of functional renal mass. Studies of the renal functional response to lithotripsy have shown that shock wave treatment triggers vasoconstriction in the kidney. This finding has been turned to advantage, and it is now known that when treatment is begun using low amplitude pulses, subsequent high amplitude shock waves are far less damaging. Thus, when shock waves are delivered judiciously, treatment can have a protective effect. The finding that cavitation is a key mechanism in vessel rupture has led to the development of novel experimental methods of shock wave delivery that can suppress bubble expansion and minimize tissue damage. Progress has also been made in understanding the physical mechanisms involved in stone comminution, and it is seen that the forces generated by cavitation, shear stress and circumferential squeezing act synergistically to fragment stones. Recent work suggests that a broad focal zone may be an advantage, allowing stones to be broken with lower amplitude pulses. Cavitation has been shown to play a critical role in reducing stone fragments to a size that can be voided. Cavitation is also the factor that limits the rate at which treatment can be performed, as stones break significantly better at slow rate than at fast ratean observation from basic research that is now appreciated in clinical practice. The current environment in lithotripsy research is encouraging. There is great interest in developing new technology, and in finding ways to improve how lithotripsy is performed. PMID- 16247351 TI - Immune response in prostate cancer. AB - Surprisingly perhaps for a tumor where there is such clear proof of a primary hormonal basis to its development, there is emerging evidence that all elements playing a key role in prostate cancer somehow affect the host immune system. This review turns the spotlight on some previously unsuspectable aspects, able to interact with the immune system in prostate cancer patients PSA, sex hormones, inflammatory infiltrates, cytokines, growth factors, neoformed blood vessels, neurotransmitters and neurotrophins, cigarette smoking, diet, therapeutic approaches. The concept of exploiting the immune system to combat cancer is not new, but only nowadays immunotherapy is a reality. Prostate cancer is an excellent target, involved in an increasing number of clinical immunotherapeutic trials. The main current purpose is to overcome the host immune tolerance of tumor cells. The most recent progresses in gene, monoclonal antibody and vaccine therapies are reported. PMID- 16247352 TI - [Zoledronic acid in the treatment of prostate cancer]. AB - Zoledronic acid is the most biologically active of all bisphosphonates. Clinical evidence suggests that it acts not only on osteoclasts and bone resorption but that it also displays antitumoral activity, proaptotic activity in synergy with antineoplastic drugs and antiangiogenic activity. Moreover, studies on zoledronic acid have reported that the agent is the only biphosphonate of demonstrated efficacy (vs. placebo) in the treatment of prostate cancer, both at a late stage of disease following bone metastasis (over 5-month delay of first skeletal event and 36% reduction in global risk of skeletal complications) and at bone demineralization in subjects receiving androgenic depletion (overall improvement of 8% in bone mineral density). PMID- 16247353 TI - Neuroendocrine differentiation after neoadjuvant hormonal treatment in prostate cancer. AB - AIM: Neuroendocrine (NE) differentiation occurs in various degree in the majority of prostatic adenocarcinomas and it has been correlated with tumor progression and poor prognosis. There is little knowledge about the impact of NE differentiation on tumor response to neoadjuvant hormonal treatment (NHT). The role of NE differentiation as a marker of recurrence after radical prostatectomy (RP) is also unclear. We evaluated whether there is an increase in NE differentiation during the course of NHT and whether the tumor relapse after radical surgery correlates with the extent of NE differentiation. METHODS: RP specimens from 44 patients submitted to 3 months of NHT and RP specimens from 40 nonpretreated patients were histologically assessed. Staining for NE differentiated prostate tumor cells was carried out using a specific monoclonal antibody against chromogranin A (CgA). RESULTS: CgA positive cells were found in 4 of 40 patients (10%) in the RP group and in 4 of 44 patients (9%) of the NHT+RP group. At follow-up, we had 21 biochemically relapsed patients. Among them, 6 were CgA positive (75% of 8 patients), whereas is were CgA negative (20% of 76 patients). CONCLUSIONS: The NE differentiation doesn't increase after NHT. Although not statistically significant a trend to higher risk of relapse among the chromogranin positive samples was observed. The significance of NE differentiation in the progression of the disease and its relation to other known prognostic factors remains unclear. PMID- 16247354 TI - Role of pharmacologic rehabilitation in the recovery of sexual function following radical prostatectomy. AB - AIM: The aim of the present study is to evaluate the role of pharmacological postoperative sexual rehabilitation in the recovery of erectile function following radical retropubic prostatectomy. METHODS: Following radical retropubic prostatectomy 113 patients were divided into 2 groups: 77 (group 1) underwent pharmacologic rehabilitation with intracavernous PGE1 starting with 5 mg once or twice weekly beginning 1 month after surgery, followed by oral sildenafil 50-100 mg twice a week after recovery of spontaneous erections, while 36 (group 2) acted as controls. The 2 groups were homogeneous for age and preoperative IIEF5 score, while nerve-sparing techniques were used more frequently in group 1. RESULTS: With 1 year follow-up 42% of group 1 patients have recovered sexual function, versus 8% of group 2 (P < 0.001). The mean recovery time was 7 months, and 56% of rehabilitated patients with bilateral preservation of the neurovascular bundles recovered sexual function, compared to 37.5% of those with unilateral preservation and 28.5% of those operated with non nerve-sparing technique. Intracavernous PGE1 caused pain in 27% of cases. CONCLUSIONS: Pharmacologic rehabilitation significantly impacts on the recovery of sexual function following radical prostatectomy, and the optimal treatment schedule is still to be defined. PMID- 16247355 TI - [Evaluation of sexual activity after prostatic surgery for benign prostatic hyperplasia assessed by pre-operative and post-operative questionnaire]. AB - AIM: Different authors showed a 5-40% incidence of erectile dysfunction after prostatic surgery for benign prostatic hyperplasia. The aim of our study was to evaluate the effect of prostatic surgery due to benign prostatic hyperplasia on sexual activity. METHODS: We evaluated 56 patients (mean age 63 years, range 49 70) who underwent transurethral resection of the prostate (30 patients, mean age 64 years, range 56-70) and open prostatectomy (26 patients, mean age 59 years, range 49-70) with a questionnaire regarding the presence of sexual dysfunction before and 3, 6, 12 months after surgery. All patients were not affected by previous or present metabolic, neurological, cardiovascular diseases. RESULTS: Twenty percent of the patients with a normal sexual activity before treatment presented erectile dysfunction after surgery. However, 35% of patients with erectile dysfunction before treatment presented an improvement of their erectile function after surgery. CONCLUSIONS: A high incidence of erectile dysfunction was observed in patients undergoing prostatic surgery. Patients with erectile dysfunction should also recover a normal behaviour after prostatic surgery as well as an improvement of the erectile dysfunction. PMID- 16247357 TI - [Primary hyperoxaluria and pregnancy]. AB - A rare case of pregnancy in a patient with primary hyperoxaluria type 1 is reported offering a clinical contribution for the prognostic study of a natural event such as the pregnancy on these patients. It is underlined how epicriticity of the hepatorenal system is remarkably altered during the last weeks of pregnancy with a great increase of calciuria and proteinuria with a risk of life for the patient and the newborn. In fact, during the 36rd week a caesarean section was performed due to gestosis. Moreover, the deterioration of kidney functionality doesn't decrease during post partum. Therefore, it would be advisable inform these patients about the risk of pregnancy. PMID- 16247356 TI - [Fluorescence endoscopy in the diagnosis and treatment of superficial bladder neoplasms]. AB - AIM: The results of a clinical investigation on neoplasm and bladder dysplasia detection by 5-aminolaevulinic acid (5-ALA)-induced fluorescence are reported. In this paper the authors report their experience with 5-ALA in the diagnosis, treatment and follow-up of bladder neoplasms after chemotherapy and endocavitary immunotherapy. METHODS: The 5-ALA was instilled in the bladder 2 h before bladder transurethral resection. This method has been used since December 2000 on 163 patients and a total of 266 biopsies were histologically examined. RESULTS: One hundred and four benign and 92 malignant/dysplastic areas were biopsied; 46 malignant/dysplastic lesions were not detected during routine white-light cystoscopy but were identified with fluorescence cystoscopy. Sensitivity was 99% but specificity was low (20%). CONCLUSIONS: ALA-based fluorescence cystoscopy is a safe and simple technique that enhances the detection of flat and papillary urothelial neoplasms. PMID- 16247358 TI - Brucella epididymoorchitis. PMID- 16247359 TI - [Mass examination in otorhinolaryngology]. PMID- 16247360 TI - [The role of rheological changes of blood and hemostasis in the development of acute neurosensory hypoacusis]. AB - We studied blood rheology and hemostasis in patients with acute neurosensory hypoacusis (ANH) of different genesis to clarify correlations of these disorders with an ANH form. After examination 72 ANH patients (60 male and 12 female, age 19-68) were divided into three groups by ANH etiology: ANH of vascular genesis (group 1, 47 patients), ANH of viral genesis (group 2, 19 patients), ANH of traumatic genesis (group 3, 6 patients). Blood rheology was studied on viscosimeter Low-Shear 30 (Switzerland); hematocrit--on Autocrit (USA) centrifuge, platelet aggregation--on aggregometer Elvi 840 (Italy), coagulation- on coagulometer Tetra (Spain). Blood rheology and hemostasis in groups 2 and 3 were changed insignificantly, while in group 1 we registered disturbed microcirculation. Treatment with vasoactive drugs normalized both clinical picture of the disease and blood rheology, hemostasis. The highest activity was seen in use of the drugs tanakan and gincor fort. PMID- 16247361 TI - [Vestibulo-ocular reflexes in patients with chronic impairment of cerebral circulation]. AB - Vestibular dysfunction was studied in 95 patients with initial cerebral circulatory insufficiency (CCI) stage I and II. Sinusoidal rotation proved highly effective in staging the disease, as initial CCI responses with vestibular hyperreflexia to sinusoidal rotation with closed eyes. With CCI progression, some patients demonstrated inhibition of vestibular excitability while other patients had enhanced excitability. This shows clinical heterogeneity of CCI stage II. P waves correlating with marked vestibulocerebellar symptoms and high vestibular excitability evidence for CCI stage II. It is established that severity of impairment of nystagmus suppression with glance fixation on a moving object depends on the disease stage. PMID- 16247362 TI - [Specific features of professional hypoacusis in airline pilots]. AB - The article deals with present-day problems of occupational hypoacusis in airline pilots. Such hypoacusis demonstrates a tendency to higher incidence. Current criteria of occupational hypoacusis for industrial workers do not satisfy the requirements of medical flight expertise as they do not consider specificities of flight conditions. This leads to erroneous expert conclusions of ENT occupational medical specialists. The problem should be solved by design of scientifically grounded criteria of assessing the state of the acoustic analyzer and flight conditions. PMID- 16247363 TI - [Papillomatosis of the lower respiratory tracts in children]. AB - Papillomatosis of the trachea, bronchi and lungs was investigated in 40 children with papillomatosis of the lower airways in comparison with 408 children with laryngeal papullomatosis. All the patients were operated in 1988-2003 with histological verification of the diagnosis. Papillomas involved the lower airways because of tracheotomy (92.5%) or tracheal intubation (7.5%). Papillomatosis of the lower airways runs a more aggressive course than laryngeal papillomatosis, its treatment efficacy is worse. All the patients suffering from laryngeal papillomatosis with a history of tracheotomy need a regular endoscopic and x-ray control as papillomatosis of the trachea and/or lungs may develop in them several years after decanulation. Pulmonary papillomatosis has an invariably poor prognosis. PMID- 16247364 TI - [The use of voicing test in phoniatrics]. AB - A study was made of the voicing-test efficacy in clinical practice. The study group consisted of 11 patients with different laryngeal diseases, the control group comprised 40 healthy subjects. Two parameters were estimated with a special computer soft: fraction of locally unvoiced pitch frames (FLUPF) and degree of voice breaks (DVB). Jitter, shimmer and signal to noise ratio were estimated as control. It is shown that the presence of locally unvoiced pitched frames is abnormal. The FLUPF test proved more precise than DVB test and, therefore, is preferential. Thus, the FLUPF test is clinically significant and can be used in clinical practice in line with parameters of acoustic analysis. PMID- 16247365 TI - [Rhinosinusogenic subdural empyema in children]. AB - We have analysed case histories of children with rhinosinusogenic intracranial complications admitted to the ENT department of the Krasnodar regional pediatric hospital for a decade (1994-2003). Of 44 patients, 10 had subdural empyema manifesting primarily with such early symptoms as severe headache (n=5), mental disorders (n=10), fever (9), focal neurological symptoms (n=10). Systemic inflammatory response syndrome occurred in all the children. Preoperative treatment included a complex of measures aimed at correction of vital functions followed by urgent surgical cleansing of the primary and secondary infection foci. Postoperative therapy in a critical unit consisted of combined antibacterial treatment (amoxicilline/clavulanate, oxacilline or vancomycin in combination with cefalosporines of the third generation and metronidasol), infusion and symptomatic therapy. Early diagnosis and treatment including eradication of the primary focus in the paranasal sinuses and drainage of the subdural abscess, antibacterial and infusion therapy provide a good outcome. PMID- 16247366 TI - [Postadenotomy soft palate paresis in children]. AB - One of frequent complications after adenotomy and tonsillectomy is paresis of the soft palate. Literature data, four groups of causes of soft palate paresis are analysed. The treatment was given to 28 patients with soft palate paresis of different etiology. Active treatment recovered function of the palate in most of the patients. PMID- 16247368 TI - [The method of conservative correction of labyrinthine disorders in operations on the ear]. PMID- 16247367 TI - [A comparative analysis of antibiotics efficacy in acute ENT infections]. AB - The comparison of the efficacy of a number of antibacterial drugs in acute pyoinflammatory ENT diseases allowed the authors to recommend moxifloxacin as most effective in acute purulent ENT pathology. Moxifloxacin demonstrated its superiority in seeding (the highest sensitivity of the bacteria to this antibiotic), in the speed of development of the bactericidal action and in treatment results. The drug had no significant side effects. PMID- 16247369 TI - [Bioparox in therapy of acute rhinosinusitis]. PMID- 16247370 TI - [Prophylaxis of iatrogenic tracheostenosis after laryngeal extirpation]. AB - Causes of such postoperative complication as cicatricial tracheostenosis after tracheotomy and laryngeal extirpation are considered. The author came to the conclusion that the leading cause of tracheostenosis is an invalid choice of tracheotomic tubes by diameter, length and angle. The experience with more than 3000 surgical interventions on the larynx and trachea allowed the author to recommend most effective treatments at each stage of cicatricial tracheostenosis. PMID- 16247371 TI - [The method of voice recovery in functional dysphonias in voice professionals]. AB - The method of the treatment of psychogenic functional dysphonia in voice professional is described. This method (audiovocalotherapy) consists in listening professional singing or speech. The mechanism of a therapeutic action of audiovocalotherapy is analysed. It is emphasized that such therapy is not expensive and can be used by ENT doctors in outpatient clinics. PMID- 16247372 TI - [Atypical course of otitis media purulenta chronica]. PMID- 16247373 TI - [Cholesteatoma of the acoustic duct]. PMID- 16247374 TI - [False aneurysm of intracavernous segment of the internal carotid artery invading the sphenoidal sinus and simulating its tumor]. PMID- 16247375 TI - [A rare case of an extensive rhinosinusogenic abscess of the brain]. PMID- 16247376 TI - [A tumor of the cochleovestibular nerve located in the depth of the auditory duct]. PMID- 16247377 TI - [Results of left atrium surgery performed during corrective surgery of mitral valve disease complicated by atriomegaly]. AB - Short- and long-term results of left atrium's plasty in 144 patients performed during correction of mitral valvular disease complicated with atriomegalia were analyzed. Age of patients ranged 7 to 66 years (33+/-9.3 ys). Size of left atrium ranged 6.0 to 9.5 cm (7.24+/-0.95 cm on average). Plastic surgery of left atrium permits to remedy symptoms of compression of left ventricle and left primary bronchus. Timely surgical treatment permits to improve central hemodynamics, to recover sinus rhythm in 25% patients, to improve quality of life in long-term period of follow-up. PMID- 16247378 TI - [Experimental validation of the method of autografts creation for arteries]. AB - Experimental studies on rats have demonstrated possibility of connective-tissue grafts formation on polyvinylchloride base introduced subcutaneously. Dynamics of autograft formation is characterized by gradual thinning of wall, regulation of collagen fibers, decreased number of cells with fibrocytes predominance. Autografts in carotid position retain its patency during 3 months, and endotheliocytes appear on their internal surface. PMID- 16247379 TI - [Sclerotherapy in patients with nodular colloid goiter]. AB - Results of percutaneous sclerotherapy with ethanol (PSTE) in 89 patients with nodular colloid goiter were analyzed. Indications and method of PSTE in different types of nodular formations were determined. All the patients were divided into 3 groups: 1st group -- 8 (9%) patients with free-functioning nodes, 2nd -- 16 (18%) patients with symptoms of cervical organs compression, 3rd -- 65 (73%) patients with cosmetic defect on neck. It is concluded that PSTE may be regarded as alternative for surgical treatment in the patients with high surgical risk. It is not pathogenic method of treatment and it should be accompanied with medicament therapy of iodine-deficient goiter. Liquidation of clinical symptoms is the criterion of treatment efficiency. PMID- 16247380 TI - [Moot points of diagnosis and treatment of hormone-producing neuroendocrine tumors of pancreas]. AB - Based on literature data and own experience of treatment of 225 patients with neuroendocrine tumors (NET) authors discuss moot points of diagnosis and treatment of this severe category of patients. It is demonstrated that the most appropriate algorithm of diagnosis before surgery is the combination of US with endoscopic US and angiography. Intraoperative diagnosis must be performed with intraoperative US and endoscopic transillumination. Authors demonstrate positive results of staged surgical treatment of MEN-1 syndrome. Malignant NET with distant metastases is not contraindication for surgical treatment. This situation is indication either for radical surgery with excision of all metastases or for cytoreductive surgery with subsequent chemoembolization and chemotherapy. PMID- 16247381 TI - [Foreign bodies of appendix and caecum complicated with acute appendicitis]. AB - Twelve patients with exogenous foreign bodies (small chicken and fish bones, portion of wire, nail, cherry stone, piece of glass) of appendix (8) or adjacent zone of caecum (4) were treated. Based on experience of treatment of these patients authors analyze pathogenesis of appendicitis associated with foreign bodies, and other aspects of this problem. PMID- 16247382 TI - [Evaluation of status severity in patients with isolated and combined injury of abdomen associated with spleen disruption]. AB - Results of treatment of 378 patients with severe combined and isolated abdominal trauma with spleen injury are analyzed. Severity of patients state and severity of trauma are assessed. General lethality was 11.9% including 7.41% during first day. Spleen-saving surgery was performed in 150 patients, 96% of them had severe injuries of organ. Spleen was removed in 228 patients. Spleen-saving surgeries (especially laparoscopic procedures) permitted to reduce significantly lethality, number of postoperative complications and hospital stay. PMID- 16247383 TI - [Influence of laser -- infrared irradiation on local complications of early postoperative period in hernias of anterior abdominal wall]. AB - Results of laser-infrared irradiation (LIRI) of postoperative wound after surgical treatment of recurrent hernias of anterior abdominal wall were analyzed. LIRI decreased number of local complications in early postoperative period in patients both with recurrent and primary hernias. The method was more effective for the treatment of local complication in patients with recurrent hernia that is confirmed by reduced of frequency of infiltrates, seromas and suppurations. PMID- 16247384 TI - [Treatment of hydatid echinococcosis of liver]. AB - Experience of surgical treatment of 373 patients with echinococcosis of liver is analyzed. Traditional surgeries were performed in 342 patients. Postoperative lethality was 1.46%, number of specific postoperative complications -- 12.5%. In long-term period recurrence was revealed in 18 (6.8%) of 264 examined patients. In 9 years after surgery residual cavities were diagnosed in 33 (9.64%) patients. Recovery was achieved in 93% patients. Experience of treatment of 31 patients with echinococcosis of liver with puncture methods (PAIR, PEVAC) was also analyzed. There were no lethal outcomes, severe complications and recurrences of disease during 2 years of follow-up. It is concluded that puncture method may be regarded as alternative for traditional surgical treatment, but it require further study. PMID- 16247385 TI - [Acute inflammation of omental processes of the colon]. AB - Results of treatment of 17 patients aged from 27 to 82 years with acute inflammation of omental processes (OP) are analyzed. The majority of them were hospitalized on day 1-3 from the beginning of the disease. In 9 cases inflammation was in the right half of the colon, in 7 -- in sigmoid colon (which in 3 patients was located in the right iliac area). In right-sided location of inflammation clinical picture looks like acute appendicitis. In some patients mainly in left-sided location of the inflammation the disease looks like renal colic, perforation of colon or peritonitis. Acute pathology of OP was suspected only in one case. All the patients underwent surgery. In 8 of them necrosis was revealed, in 7 -- phlegmonous inflammation of one OP, in 2 -- inflammatory infiltration without destruction. Surgery comes to removing of affected OP, which was supplemented with appendectomy in 9 cases. There were no complications and lethal outcomes. Problems of diagnosis and surgical treatment of acute inflammation of OP are discussed. PMID- 16247386 TI - [Mistakes in surgical treatment of generalized purulent peritonitis]. AB - Classification of mistakes in technique and tactics in the treatment of acute generalized purulent peritonitis is developed. All the mistakes were divided into two groups -- technical and tactical -- before, during and after surgery. This classification permits to understand their concrete role in pathogenesis of complications and causes of lethal outcomes. PMID- 16247387 TI - [Dental enamel structure and its relation to dentine]. AB - New information on microscopic and ultrastructural architectonics of human dental enamel and its relation to dentin is presented. Enamel consolidation with dentin is mediated by borderline layer of calcificated substance having fibrous structure. PMID- 16247388 TI - [Effects of the method of mechanical root canal treatment on emergence of pain after endodontic management]. AB - For assessment of tooth canal mechanical preparation method influence upon frequency of pain emergence after endodontic treatment, laboratory and clinical evaluation of sound tools of the Sonic system, ultrasound tools of the Satelec Suprasson system, full-wind tools of Pro Taper and System GT as well as handy K files was performed. It was found that the least risk of pain emergence after endodontic treatment occurs with tooth canal widening by CrownDown technique because in this case the minimal amount of canal preparation products are educted into periodontal tissues. PMID- 16247389 TI - [Gingival tissue blood circulation in health and parodontal diseases as assessed by ultrasonic Dopplerography]. AB - Hemodynamic indices of blood circulation in gingival tissues in health and parodontal diseases were determined basing on ultrasonic Dopplerography data on linear and volumetric blood circulation flow rate. It is shown that changes in hemodynamics of tissual blood flow in initial inflammatory diseases are moderate (linear and volumetric blood circulation flow rate came down by 22-40%). In more severe inflammation in the parodontium and in cases of parodontosis changes in hemodynamics of tissual blood flow were more pronounced (linear and volumetric blood circulation flow rate came down by 56-80%). PMID- 16247390 TI - [Modern approach to pathogenetic therapy of the oral mucosa lichen]. AB - Study of humoral immunity, morphofunctional status of cell factors of immunologic reactivity (T-lymphocytes, neutrophils) and intestinal microecology allowed design of comprehensive pathogenetic therapy of oral lichen. PMID- 16247391 TI - [Characteristics of caries and parodontal diseases in young men with constitutionally determined nutrition insufficiency]. AB - 151 young men with constitutionally determined nutrition insufficiency were examined. For absolute majority of them multiple caries with primary lesions development in incisors and on 3 molar surfaces was typical. In patients with constitutionally determined nutrition insufficiency caries complications with predominance of acute apical or chronic periodontitis were 3 times more often diagnosed. Low level of oral hygiene was noted in such young men that correlated with high prevalence of severe gingivitis in them. PMID- 16247392 TI - [Mandibular contracture as a complication of trigeminal third branch anesthesia]. AB - Wide use of local trigeminal third branch anesthesia not rarely leads to mouth opening restriction. There is no common opinion on etiology and pathogenesis of this complication, that explains also the absence of treatment algorithm for such patients depending upon length of the process development. PMID- 16247393 TI - [Experimental and clinical substantiation of allogenic cortical bone matrix use for treatment of patients with habitual mandibular dislocation]. AB - Cortical bone matrix (CBM) was fixed under the zygomatic arch of 20 experimental rabbits by the titanium wire. CBM transformed into the organotypic bone tissue during 90-180 days, and the transformation processes were more significant if the cortical layer of the inferior border of the zygomatic arch had been removed and CBM had been miotisized. Taking into account good results of the experiment, CBM was applied in clinic for treatment of habitual mandibular dislocation (HMD). The new operative approach to the TMJ and method for HMD using CBM was developed. It is easy to perform, less traumatic and provides reliable long-term results. Of 36 operated joint only one recurrence of the dislocation was observed. PMID- 16247394 TI - [Variants of surgical interventions in cases of a craniofacial form of fibrous dysplasia]. AB - From 1980 till now we observed 6 cases of fibrous dysplasia primarily in patients of an older age group with a long history of the disease. Indications to operative interventions were such increasing clinical manifestations as functional derangements, progressing pain syndrome, suspected malignant transformation. In 1 patient with fibrous dysplasia after carried out earlier radiotherapy we observed malignant transformation into sarcoma with process progressing and lethal outcome. Variants of extended operative interventions in cases of craniofacial form of fibrous dysplasia are discussed as well as modern techniques of reconstruction of such defects of craniofacial zone. PMID- 16247395 TI - [About surgical treatment of mucous membrane of mouth floor and tongue cancer]. AB - In the department of head and neck tumors of Rostov Research Oncological Institute the operation on advanced mouth floor and tongue cancer is proposed comprising removement of the regional lymph nodes and resection of the tongue and mouth floor. 30 patients were operated by the suggested technique with restoration of the functions of swallowing, nutrition and speech in postoperative period in all cases. 3-year recurrence-free survival made up 28.4+/-8.2%. PMID- 16247396 TI - [Experimental substantiation of parameters and strength characteristics of new endodontic-endoosseous implant construction]. AB - Postoperative tooth stump is not able to endure usual functional loads adequately that leads to enhanced resorption of bone tissues surrounding root and loss of the tooth. Based on math computer simulation of mode of deformation of biomechanical model "metal-ceramic construction--endodontic-endoosseous implant (EEI)--stump of operated tooth--bone tissue of the jaw" new domestic EEI parameters were elaborated which improved greatly biomechanical properties of tooth with operated roots. Strength characteristics of new EEI were studied. A set of instruments for EEI was developed. PMID- 16247397 TI - [Orthodontic service as one of the factors of improving quality of life]. AB - Opinion of the population of 4 regions of Russia on orthodontic treatment was studied basing upon received questionnaire data analysis. Among the questions- reasons, age at the moment of orthodontist addressing, estimate of treatment potentials, attitude to chargeable treatment. PMID- 16247398 TI - [Angle class I malocclusion in primary dentition and findings in permanent dentition--a follow-up study]. AB - The aim of the study was to examine stability and changes in Angle Class I malocclusion from deciduous to permanent dentition in 168 subjects. All the subjects had Class I malocclusion in deciduous dentition, and were examined by the same orthodontist on two occasions during deciduous and permanent dentition. None of the subjects had received orthodontic therapy in the meantime. The results showed considerable changes from primary to permanent dentition. Crowding in primary dentition was retained in permanent dentition in 45.2% cases. In 16.2% cases it changed into normocclusion and 38.6% subjects developed other types of malocclusion. Open bite was retained in permanent dentition in 17.8% cases and in 17.8% subjects transformed into normocclusion. 64.4% subjects developed other types of malocclusion. Cross bite was retained in permanent dentition in 21.4% cases and in 28.6% subjects changed to normocclusion. Other types of malocclusion in permanent dentition developed in 50% subjects. In 30.8% of cases finding of premature loss of deciduous teeth was accompanied by extraction of some permanent teeth. Normocclusion was retained in 19.2% cases while 50% of children developed some type of malocclusion. Crowding, which was retained in permanent dentition in 45.2% cases, showed the highest degree of stability. Children with this type of anomaly in primary dentition displayed the highest frequency of total malocclusions (83.3% subjects). Out of all anomalies in primary dentition, cross bite most frequently switched to normal occlusion in permanent dentition (in 28.6% cases). PMID- 16247399 TI - [Modification of Svenson's biopsy of rectal wall in diagnosis of malformations of intramural nervous system in adults]. AB - Thirty-five autopsied specimens of rectum's distal part and anal canal part were studied. It was revealed that length of physiological hypogangliosis zone ranged from 7.5 to 50.0 mm (mean 24.4+/-10.9 mm). With regard to this wide range the modification of transanal Svenson's biopsy of rectal wall was proposed for diagnosis of malformations of intramural nervous system of the colon. The method was used in 21 patients with megacolon. The results demonstrate accuracy and safety of this diagnostic method. PMID- 16247400 TI - [Urgent resection of the colon with compressive anastomosis and afferent colostoma]. AB - Thirty patients with colon disease in need of urgent surgery have undergone resection of the colon with formation of terminal afferent colostomy and compressive colon-colon anastomosis using oval and linear nikelid-titanium implants. Complications were seen in 6 (20%) patients, lethality was 6,7% due to multiorgan failure in the presence of generalized peritonitis. There were no complications due to colon-colon anastomosis. The above method permits one to reduce surgical trauma of future reconstructive surgeries. PMID- 16247401 TI - [Prosthesis of the aortal valve in adult patients with "narrow" aortic root]. AB - Results of treatment of 54 patients with aortal valvular disease and "narrow" aortic root are analyzed. Only adult patients were included in the study, their age ranged from 16 to 64 years (mean age 41.5+/-14.7 yrs). Thirty-four (63%) patients of them have undergone aortic valve replacement with disk prosthesis, in 20 (37%) patients this surgery was supplemented with dillative plasty of fibrous ring and aortic root by Nicks's method. The proposed algorithm of choosing the most effective operation in patients with the aortal defect and narrow aortal root permits one to avoid pathologic pressure gradient between the left ventricle and the aorta after surgery and to achieve the most favorable clinical effect. PMID- 16247402 TI - [Flap transverse sternotomy during operations on the vessels and organs of the upper mediastinum and the first cervical zone]. AB - A surgical approach to vessels and organs of the upper mediastinum was grounded, developed and tested. An experimental study of flap transverse sternotomy was carried out in 20 cadavers. A wide approach with throwing of the presternum was the best for manipulations in proximal parts of the aortic branches. This approach was also less traumatic. PMID- 16247403 TI - [Surgical treatment of patients with chronic lower limb venous insufficiency and trophic disorders]. AB - The experience in surgical treatment of 326 patients with IV-VI degree chronic lower limb venous insufficiency (CEAP) is analyzed. Correction of disorders of venous hemodynamics was performed with surgical methods and sclerotherapy. The best clinical and functional results in long-term period were achieved in patients undergone surgery in an early stage of the disease without deep veins valves insufficiency and external trophic disorders. Pathology of deep veins required correction of valves function. It is concluded that these patients require complex approach to the treatment. PMID- 16247405 TI - [Local plastic surgeries in the treatment of total burn cicatrical contractures of the neck]. AB - Results of treatment of 44 patients with total burn cicatrical contractures of the neck were analyzed. In all the cases deformation of the neck was corrected by plasty with contradirectional myofascial flaps from cicatrical tissues. Clinical characteristics of burn cicatrices helped determine criteria for their using in plastic surgery. The method may be used both as a final variant of neck plastic repair and as a preparatory stage before other methods of surgical reconstructions. PMID- 16247404 TI - [Prophylaxis and treatment of disorders of the vocal cords mobility after surgeries on the thyroid gland]. AB - Own and foreign experience in surgical treatment of thyroid gland (TG) diseases was analyzed. Surgical anatomy of TG, effective methods of surgical techniques, classification of disorders of vocal cords mobility, and treatment policy are described. It is demonstrated that no less than 70-80% patients with respiratory disorders after surgeries on TG can avoid tracheostomy and subsequent complex and sometimes staged plastic surgeries on the larynx. Proposed recommendations permit one to reduce risk of postoperative complications, to improve long-term results of surgical treatment and life quality of operated patients. PMID- 16247408 TI - [Surgeries on gastrointestinal tract without components of donor's blood]. AB - Results of surgical treatment without transfusion of donor's blood in 58 patients were compared with ones in 40 patients treated with allogenous transfusions. Blood-saving program included preoperative storage of autoplasma, acute normovolemic hemodilution, recombinant human erythropoietin and perftoran. Both groups of patients were similar by types of surgeries performed on gastrointestinal tract. Results of treatment in the study group (58 patients) were better: there were less postoperative complications; period of postoperative rehabilitation was shorter. It is demonstrated that it is possible to refuse allogenous hemotransfusion in 96.6% cases of traumatic surgeries on gastrointestinal tract. PMID- 16247407 TI - [Focal cirrhosis of the liver]. AB - The experience in treatment of 14 patients with focal cirrhosis of the liver is summarized. Clinical and morphological data is presented. It is noted that clinical picture is poor and non-specific. Ultrasonic examination is the main diagnostic tool. It is demonstrated that histological examination of removed specimen is often necessary for final diagnosis. Method of choice in the treatment of patients is surgical one. The scope of surgery depends on intraoperative features, extended hepatic resection is not usually indicated. PMID- 16247406 TI - [Comparative analysis of different methods in the treatment of patients with acute calculous cholecystitis and choledocholithiasis]. AB - In the patients with different forms of cholelithiasis 1988 operations were performed, 597 of them - on account of acute calculous cholecystitis. In 118 (19%) patients with acute calculous cholecystitis choledocholithiasis was diagnosed. All the patients were divided into two groups. Group 1 consisted of 52 (44%) patients who have undergone conventional "open" operation with choledocholithotomy. Group 2 consisted of 66 (56%) patients treated with endoscopic techniques. In group 1 complications were seen in 4 (7.6%) patients, lethal outcome -- in 3 (5.7%), residual choledocholithiasis -- in 7 (13.4%). Mean hospital stay was 24.7 days. In group 2 complications were seen in 1 (2%) patient, residual choledocholithiasis -- in 1 (2%), there were no lethal outcomes. Mean hospital stay was 15.1 days. It is concluded that minimally invasive technologies in the treatment of acute calculous cholecystitis and choledocholithiasis improve results of treatment and shorten hospital stay. PMID- 16247409 TI - [Organ-saving method of treatment of spleen disruption]. AB - Based on 102 X-ray and 73 anatomic examinations, the scheme of arterial circulation of the pancreas and spleen, features of their collateral circulation are analyzed. Optimal level of spleen artery ligation at proximal part as the main stage of organ-saving surgical treatment in traumatic disruption of the spleen is substantiated. PMID- 16247410 TI - [Adequate gas exchange in two-sided and combined surgeries on thoracic and abdominal organs]. AB - Method of combined artificial lung ventilation during simultaneous two-sided operations on the lungs was studied in 60 patients. This method is based on traditional ALV with parameters calculated according to functional state of the patient's respiratory system, and additional high-frequency jet pulmonary ventilation. The developed method was not accompanied by any severe disorders of gas exchange and mechanics of ventilation. PMID- 16247411 TI - [Diagnostic and therapeutic treatment policy in ulcerous gastroduodenal bleedings]. AB - Results of treatment of 796 patients with ulcerous gastroduodenal bleedings (UGDB) are analyzed. The study group consisted of 676 patients who have undergone endoscopic hemostasis. The control group consisted of 120 patients treated with traditional therapy. Endoscopic hemostasis in the study group was effective in 628 (92.8%) patients, recurrence of bleeding was seen in 21 (3.3%) patients. Effective hemostasis in the control group was achieved in 92 (76.6%) patients, recurrence of bleeding occurred in 13 (14.1%) patients. Positive result of Helicobacter pylori detection was in 600 (88.7%) patients. It is concluded that local treatment of ulcer with glue applications reduces 3 times number of bleeding recurrences. PMID- 16247413 TI - Unrelated donor bone marrow transplantation with treosulfan-based myeloablative conditioning for paroxysmal nocturnal hemoglobinuria--successful treatment despite multiple transplant-related risk factors for hemolysis including major Kidd group incompatibility. PMID- 16247412 TI - Caffeic acid phenethyl ester modulates Helicobacter pylori-induced nuclear factor kappa B and activator protein-1 expression in gastric epithelial cells. AB - Caffeic acid phenethyl ester (CAPE), an active component of propolis from honeybee hives (honeybee resin), has anti-inflammatory, anti-carcinogenic and anti-bacterial properties. This study was designed to investigate the anti inflammatory effects of CAPE on Helicobacter pylori-induced NF-kappaB and AP-1 in the gastric epithelial cell line AGS. Electrophoretic mobility shift assay was used to measure NF-kappaB- and AP-1-DNA binding activity. Western blotting was used to detect IkappaB-alpha and COX-2 expression in AGS cells cocultured with H. pylori. The antiproliferative effect of CAPE was measured by MTT assay. Our results showed that caffeic phenethyl ester inhibits H. pylori-induced NF-kappaB and AP-1 DNA-binding activity in a dose (0.1-25 microg ml(-1) approximately 0.35 88 microM) and time- (15-240 min) dependent manner in AGS cells. Maximum inhibition by CAPE was observed at concentrations of 25 microg ml(-1) ( approximately 88 microM) CAPE prevented H. pylori- and cytokine-induced degradation of IkappaB-alpha protein. Pretreatment of AGS cells with CAPE also blocked cytokine- and mitogen-induced NF-kappaB and AP-1 expression. Furthermore, CAPE suppressed H. pylori-induced cell proliferation and production of the cytokines TNF-alpha and IL-8. In addition, CAPE blocked H. pylori-induced COX-2 expression. The inhibition of such transcription by CAPE could result in suppression of many genes during H. pylori-induced inflammation, and also provide new insights into the anti-cancer and anti-inflammatory properties of CAPE. PMID- 16247414 TI - Another case of PORN (bilateral progressive outer retinal necrosis) after allogeneic stem cell transplantation. PMID- 16247415 TI - Sequential, cycling maintenance therapy for post transplant multiple myeloma. AB - High-dose chemotherapy with autologous stem cell transplantation in patients with newly diagnosed multiple myeloma can prolong survival but is not curative. Maintenance therapy post transplant may prolong the disease-free interval and impact overall survival. We have conducted a phase II pilot study of 28 post transplant myeloma patients treated with a sequential, cycling maintenance regimen. The regimen was designed to include a variety of active myeloma agents chosen for ease of administration to enhance patient compliance and scheduled sequentially to minimize toxicity. The 12-month cycling schedule included dexamethasone (months 1-3); melphalan and prednisone (months 4, 5); cyclophosphamide and prednisone (months 6, 7); alpha-interferon (months 8-10); followed by a drug holiday (months 11, 12). The regimen was generally well tolerated with five patients developing reversible grade III-IV toxicity (diabetes-induced hyperglycemia in four, neutropenia in one). There was one toxic death on study due to non-neutropenic pneumonia and sepsis. Median event-free survival from transplant was 36.9 months (95% CI 23.6 - upper limit not yet reached) with median overall survival not yet reached at a median follow-up of 44 months. This concept of cycling, sequential maintenance with various agents, perhaps including newer biological, targeted agents, warrants further investigation in multiple myeloma. PMID- 16247416 TI - Allogeneic stem cell transplantation in children with acute lymphoblastic leukemia after isolated central nervous system relapse: our experiences and review of the literature. AB - The prognosis of patients with acute lymphoblastic leukemia (ALL) and central nervous system (CNS) relapse has historically been very poor. Although chemo radiotherapy has improved outcomes, some patients still have a poor prognosis after CNS relapse. Therefore, allogeneic hematopoietic stem cell transplantation (allo-SCT) has recently become an option for treatment of CNS leukemia; however, information, particularly on the long-term outcome of transplant recipients, is limited. We performed allo-SCT in eight pediatric patients with ALL (n=7) or T cell type non-Hodgkin's lymphoma (n=1), who had isolated CNS relapse. All patients survived for a median of 70.5 (range, 13-153) months after SCT. Sequelae developed late in some patients: mental retardation (IQ=47) in one patient, severe alopecia in two patients, limited chronic graft-versus-host-disease in three patients, and amenorrhea and/or hypothyroidism in three patients. Except for a pre-school child with post transplant CNS relapse, six out of seven patients show normal school/social performance. Our results clearly indicate a high cure rate of isolated CNS relapse by allo-SCT in pediatric lymphoid malignancies; however, there needs to be further studies to determine which are the appropriate candidates for transplantation and what is the best transplant regimen to achieve high cure rate and maintain good quality of life. PMID- 16247417 TI - Daclizumab for children with corticosteroid refractory graft-versus-host disease. AB - Daclizumab, a humanized IL-2 receptor antagonist, has been found to be safe and effective in adults with refractory graft-versus-host disease (GVHD); however, data describing its efficacy for refractory GVHD in children are limited. We report a series of 14 children who were treated with daclizumab for severe acute and/or chronic corticosteroid refractory GVHD. Patients were treated with 2 mg/kg weekly for 8 weeks followed by 1 mg/kg weekly for 4 weeks. Nine of 14 patients responded to daclizumab as measured by improvement of GVHD symptoms, and the ability to substantially wean corticosteroid dose. Five of 11 patients with acute GVHD had complete symptom resolution, and 2/11 had a partial response. Two of four patients with chronic GVHD had complete symptom resolution. In these patients, daclizumab was only effective in treating skin GVHD. Seven of the nine patients who had a complete or partial response eventually had recurrence of GVHD; however, the GVHD was less severe and no longer corticosteroid refractory. There was no infusional toxicity, and no infections that could be attributable to the drug. Daclizumab is a relatively safe and effective medication for corticosteroid refractory GVHD in children and larger studies are needed to evaluate its role in treatment. PMID- 16247418 TI - Low CXCR4 membrane expression on CD34(+) cells characterizes cells mobilized to blood. AB - SUMMARY: In this work, association between the presence and membrane density of CXCR4 and the effectiveness of mobilization was studied. Ninety G-CSF mobilized PBPC and 28 native BM (nBM) preparations obtained from healthy individuals for transplantation and BM obtained after G-CSF mobilized PBPC collection in 10 donors were investigated. Positivity for CD34, HLA-DR and CXCR4 were analysed in the three colour fluorescence. The cellular profile of PBPC differed from nBM preparations with respect to lower: (i) proportion of CD34(+) cells (0.64%+/-0.04 vs 0.92+/-0.07) and CD34(+)CXCR4(+) cells (0.30%+/-0.02 vs 0.61+/-0.07); (ii) contribution of CXCR4(+) cells to CD34(+) cells (52.2%+/-2.5 vs 62.2%+/-4.2); (iii) CXCR4 epitope density in CD34(+) cells (48.9+/-5.5 vs 94.7+/-10.4). PBPC yield for CD34(+) cells was correlated with the content of CD34(+) cells lacking CXCR4 in the leukapheresis product (R=0.38). In contrast, nBM harvested for transplantation was poor in CD34(+) cells if these cells were frequently CXCR4- (R=-0.49). The present study shows that CD34(+) cells mobilized to blood were characterized with a low proportion of CXCR4 and this associated with CD34(+) cell content in PBPC. PMID- 16247419 TI - Bone marrow transplantation from unrelated donors for patients with adult T-cell leukaemia/lymphoma. AB - Adult T-cell leukaemia/lymphoma (ATLL) is a highly aggressive haematological malignancy. More than 40 cases of ATLL treated by allogeneic bone marrow transplantation (BMT) from sibling donors have been reported, while there have been only a few cases of unrelated BMT for treatment of this disease. We began performing allogeneic BMT from unrelated donors in 1999 to improve the outcome of ATLL patients with no suitable sibling donors. Eight ATLL patients underwent unrelated BMT; five received the conventional conditioning regimen consisting of cyclophosphamide and total body irradiation, while three received a reduced intensity preparative regimen. Two patients died due to encephalopathy of unknown aetiology on days 10 and 35, and one patient died due to progression of ATLL 25 months after BMT. Five patients are currently alive and disease-free at a median of 20 months after BMT. Proviral human T-lymphotropic virus type-I (HTLV-I) DNA load in peripheral blood mononuclear cells (PBMCs) was assessed in four cases before and after BMT. HTLV-I proviral DNA load was reduced significantly after transplantation. Unrelated BMT is feasible for treatment of ATLL. Further studies in a larger number of cases are required to determine the optimal conditioning regimen and stem cell source. PMID- 16247420 TI - Effects of various antireabsorptive treatments on bone mineral density in hypogonadal young women after allogeneic stem cell transplantation. AB - Ovarian failure after allogeneic stem cell transplant (allo-SCT) is an important risk factor for development of osteoporosis. We investigated the effects of various antiresorptive treatments in long-term surviving females with ovarian failure after allo-SCT. A total of 60 women with osteoporosis or osteopenia were divided randomly into four groups of 15 women each. Group 1 was treated with calcium and vitamin D alone, group 2 received the same treatment in combination with hormone replacement therapy (HRT), group 3 received risedronate (35 mg weekly, orally for 1 year) and group 4 zoledronic acid (3 monthly doses of 4 mg (intravenous)). All groups were similar for age, body mass index, underlying disease and time elapsed from transplant. Lumbar and femoral bone mineral density (BMD) were measured at baseline and after 12 months, together with serum osteocalcin and urinary hydroxyproline. At 12 months, a significant decrease in lumbar and femoral BMD was observed in group 1 and a milder decrease in group 2. Risedronate treatment increased significantly lumbar BMD and prevented bone loss at the femoral neck. Zoledronic acid increased significantly both lumbar and femoral BMD. In groups 3 and 4 the hydroxyproline excretion was significantly reduced, while osteocalcin mildly increased only in group 4. In conclusion, bisphosphonate administration is useful to prevent and treat bone demineralization in young adult women after allo-SCT. PMID- 16247421 TI - Donor age and degree of HLA matching have a major impact on the outcome of unrelated donor haematopoietic cell transplantation for chronic myeloid leukaemia. AB - We analysed the outcome of 92 consecutive unrelated donor haematopoietic cell transplantations (UD-HCTs) performed in Spain to treat adult patients with CML in the first chronic phase (1CP). Patients' and donors' median age was 32 (15-49) and 36 (22-56) years, respectively. In all, 73 pairs (79%) matched for A, B+/-C and DRB1+/-DQB1 loci and 19 had > or =1 mismatch. Their probability of survival and disease-free survival at 4 years were 50 and 46%, respectively. Pretransplant factors associated with a better survival were patient age <25 years (P=0.035), donor age < or =36 years (P=0.012), use of cyclosporine since day -7 (P=0.001), and matching 8/8, 9/10 or 10/10 loci at allele level (P=0.003). In multivariate analysis only donor age (P=0.003; RR=3.1 (95% CI: 1.3-7.1)) and degree of HLA matching (P=0.009; RR: 7.7 (95% CI: 1.8-33)) maintained their significance. The addition of these two variables to the EBMT prognostic score allowed an adequate risk assessment for patients receiving a UD-HCT during 1CP. Our analysis shows that in patients with a young and fully allele-matched donor, UD-HCT should be considered in the initial therapeutic algorithm due to its excellent outcome (92% survival at 2 years). PMID- 16247422 TI - Autologous stem cell transplantation followed by consolidation chemotherapy for patients with multiple myeloma. AB - Although high-dose therapy and autologous stem cell transplant (ASCT) is superior to conventional chemotherapy for treatment of myeloma, most patients relapse and the time to relapse depends upon the initial prognostic factors. The administration of non-cross-resistant chemotherapies during the post-transplant period may delay or prevent relapse. We prospectively studied the role of consolidation chemotherapy (CC) after single autologous peripheral blood stem cell transplant (auto-PBSCT) in 103 mostly newly diagnosed myeloma patients (67 patients were < or =6 months from the initial treatment). Patients received conditioning with BCNU, melphalan+/-gemcitabine and auto-PBSCT followed by two cycles of the DCEP+/-G regimen (dexamethasone, cyclophosphamide, etoposide, cisplatin+/-gemcitabine) at 3 and 9 months post-transplant and alternating with two cycles of DPP regimen (dexamethasone, cisplatin, paclitaxel) at 6 and 12 months post-transplant. With a median follow-up of 61.2 months, the median event free survival (EFS) and overall survival (OS) are 26 and 54.1 months, respectively. The 5-year EFS and OS are 23.1 and 42.5%, respectively. Overall, 51 (49.5%) patients finished all CC, suggesting that a major limitation of this approach is an inability to deliver all planned treatments. In order to improve results following autotransplantation, novel agents or immunologic approaches should be studied in the post-transplant setting. PMID- 16247423 TI - Herpes zoster infection in the post-hematopoietic stem cell transplant pediatric population may be preceded by transaminitis: an institutional experience. AB - Herpes zoster (HZ), a varicella-zoster virus reactivation, frequently complicates hematopoietic stem cell transplantation (HSCT). Its incidence, complications, and associated risk factors in 310 children undergoing HSCT were reviewed. In all, 61 of 201(32%) patients who had undergone allogeneic and 10 of 109 (9%) patients who had undergone autologous HSCT developed HZ. Of 90 VZV seropositive allogeneic patients, 50 (53%) developed HZ. Seven (17%) of 41 VZV seropositive autologous patients developed HZ. Although a substantial number of patients develop HZ in the early post-HSCT period, risk for HZ persists and HZ can occur up to 5 years post-HSCT. Risk factors for HZ included age >10 years (P<0.0001), allogeneic HSCT (P<0.001), and total body irradiation (TBI) (P<0.059) in allogeneic recipients. Of 37, 22 (59%) patients experienced an elevated alanine aminotransferase (ALT), unassociated with GVHD, in the month preceding HZ. Of the 48/64 patients (75%) hospitalized for treatment (median stay, 6 days; range, 2-39), length of stay was unaffected by donor type but increased by cutaneous dissemination and visceral involvement (P=0.023 and 0.034, respectively) in allogeneic patients. Consideration of HZ infection particularly in patients >10 years of age with elevated ALT after TBI-conditioned allogeneic HSCT may permit earlier diagnosis and therapeutic intervention. PMID- 16247424 TI - The early referral for reduced-intensity stem cell transplantation in patients with Ph1 (+) chronic myelogenous leukemia in chronic phase in the imatinib era: results of the Latin American Cooperative Oncohematology Group (LACOHG) prospective, multicenter study. AB - Using a reduced-intensity stem cell transplantation (RIST) schedule, 24 patients with Philadelphia (Ph1) (+) chronic myelogenous leukemia (CML) in first chronic phase (CP) were prospectively allografted in four Latin American countries: Mexico, Brazil, Colombia and Venezuela, using HLA-identical siblings as donors. The median age of the patients was 41 years (range 10-71 years); there were eight females. Patients received a median of 4.4 x 10(6)/kg CD34 cells. The median time to achieve above 0.5 x 10(9)/l granulocytes was 12 days, range 0-41 days, and the median time to achieve above 20 x 10(9)/l platelets was also 12 days, range 0-45 days. In all, 22 patients are alive 81-830 (median 497) days after RIST. The 830 day probability of survival is 92%, and the median survival has not been reached, being beyond 830 days. A total of 11 patients (46%) developed acute graft-versus host disease (GVHD), and seven of 23 (30%) developed chronic GVHD. Two patients died 43 and 210 days after RIST, one as a result of sepsis and the other of chronic GVHD. The 100-day mortality was 4.4%, and transplant-related mortality was 8%. RIST for patients with CML in CP appears to be an adequate therapeutic option. PMID- 16247425 TI - High-resolution HLA typing by sequencing for HLA-A, -B, -C, -DR, -DQ in 122 unrelated cord blood/patient pair transplants hardly improves long-term clinical outcome. AB - To determine the impact of high-resolution (HR) HLA typing with outcomes after UCBT, DNAs of 122 pairs were analysed for HLA class I and class II mismatches (MM) based on HR typing. For HLA-A, -B on low-resolution typing and -DRB1 on HR typing, the following MM situation resulted: no MM (13%), one MM (40%), two MM (36%), three MM (8%), four MM (3%). For A, B, C, DR and DQ based on HR typing the following MM occurred: No MM (4%), one MM (10%), two MM (15%), three MM (22%), four MM (25%), five MM (12%), six MM (6%), seven MM (3%), eight MM (2%). There was no significant association between number of MM (HR) for both HLA-A, -B and DRB1 and HLA-A, -B, -C, -DRB1 and DQB1 and aGvHD grade II-IV. There was a trend that MM in class I HR were associated with neutrophil recovery; HLA-A locus typing analysed in HvG direction was associated with reduced cumulative incidence of engraftment (P=0.04), the same for C-KIR in HvG direction (P=0.01). No significant correlation was found between numbers of HLA-MM on the HR level with 2-year survival. The analysis shows that the degree of mismatching in UCBT is even higher than expected. PMID- 16247426 TI - Successful management of sudden, massive immune hemolysis following ABO incompatible allogeneic PBSCT despite methotrexate GvHD prophylaxis. PMID- 16247427 TI - Acute, life-threatening hypoglycemia associated with haloperidol in a hematopoietic stem cell transplant recipient. PMID- 16247428 TI - A prospective randomized study on the mobilization of CD34+ cells comparing continuous intravenous vs subcutaneous administration of rhG-CSF in normal donors. AB - The efficacy of mobilizing peripheral blood progenitor cells (PBPC) with continuous intravenous (c.i.v.) administration of rhG-CSF was randomly compared to subcutaneous (s.c.) administration, in 15 normal donors in each arm of the study for 6 days. The percentage and absolute numbers of CD34+ cells in the c.i.v. and s.c. groups increased maximally at day 3 and 5, respectively, when compared with the steady-state (day 0) level. Peak CD34+ cell levels were achieved on day 3 in the c.i.v. group, with more rapid results than in the s.c. group (49.3/microl vs 35.9/microl, P=0.043). Plasma rhG-CSF levels declined progressively during mobilization in each group as the WBC increased. The serum level of rhG-CSF did not correlate with CD34+ cell counts in the peripheral blood. Toxicity profiles in the c.i.v. and s.c. groups were similar. Each regimen was effective in successfully mobilizing the target CD34 cell number. PMID- 16247430 TI - Human herpesvirus 6 variant A infection with fever, skin rash, and liver dysfunction in a patient after unrelated cord blood transplantation. PMID- 16247429 TI - Rapid response to alefacept given to patients with steroid resistant or steroid dependent acute graft-versus-host disease: a preliminary report. AB - We evaluated the effect of alefacept (Amevive), a novel dimeric fusion protein, in steroid resistant/dependent acute graft-versus-host-disease (aGVHD). Seven patients were treated in eight aGVHD episodes. GVHD grade at treatment initiation and at peak ranged 2-4 (median 2.5) and 2-4 (median 4), respectively. System involvement at GVHD peak included skin (n=7), gastrointestinal tract (n=5) and liver (n=3). All patients responded. However, one patient with skin GVHD and two with gastrointestinal GVHD featuring an early initial response (IR) exacerbated and CR was not achieved. Skin GVHD responded rapidly with a median of 1 day to IR and 7 days to CR. Intestinal response was slower with median 7.5 days to IR. Of the four patients that achieved IR, CR was achieved in only one (40 days to CR). None of the patients had significant hepatic GVHD before treatment so no hepatic effect of alefacept could be determined. No immediate alefacept-related side effects were observed. Late side effects included infections (aspergillus sinusitis, pneumonia, bacteremia, pharyngeal thrush), pancytopenia and hemorrhagic cystitis. Three patients had CMV reactivation while on alefacept. We conclude that alefacept may have a beneficial effect in controlling aGVHD. Further investigations in larger cohorts of patients and controlled studies are warranted. PMID- 16247431 TI - Dendritic cells in autologous hematopoietic stem cell transplantation for diffuse large B-cell lymphoma: graft content and post transplant recovery predict survival. AB - Allograft dendritic cell (DC) content has been identified as a predictor of relapse and event-free survival after allogeneic bone marrow transplantation. However, the prognostic importance of DCs has not been evaluated in the setting of autologous hematopoietic stem cell transplantation (HSCT). We prospectively determined pre-transplant and post transplant DC levels, including DC1 and DC2 subset levels, in 53 patients with diffuse large B-cell non-Hodgkin's lymphoma (DLBC NHL) undergoing autologous HSCT. Pre-transplant DCs were measured in the collected stem cell products and were therefore indicative of cell numbers infused directly into patients; post transplant analysis of DCs was performed on the peripheral blood of patients 6 weeks after the infusion of autologous stem cells. Higher pre-transplant levels of DC1 cells and total DCs were significantly associated with improved survival. Similarly, greater post transplant levels of total DCs and both subsets were significantly associated with survival. These findings suggest a relationship between DC reconstitution and survival following autologous HSCT for DLBC NHL. Strategies to increase autograft DC content or accelerate DC recovery after autologous HSCT might improve outcomes in this setting. PMID- 16247432 TI - Radioimmunoconjugates in acute leukemia treatment: the future is radiant. AB - Targeted radiotherapy of the bone marrow using radiolabeled monoclonal antibodies is a therapeutic approach of considerable potential for the treatment of acute leukemia in addition to or as a substitute for total body irradiation. The data currently available, of about 300 patients, suggest that radioimmunotherapy (RIT) with beta-emitters in acute leukemia is feasible and safe using a variety of antibodies (anti-CD33, anti-CD45, anti-CD66) and radionuclides (131I, 90Y, 188Re). It appears to reduce the risk of relapse in high-risk acute myelogenous leukemia (AML) patients transplanted early in the course of their disease (<15% blasts) to 20-30%. Furthermore, it has shown the potential to safely intensify reduced-intensity conditioning regimens (nonrelapse mortality of 25% compared to relapse rate of 55% within 2 years). Significant improvements in the results of refractory patients will probably depend on the successful further development of RIT with alpha-emitters or the use of a cocktail of antibodies labeled with alpha and beta-emitters, in a first dose escalation study of 213Bi-labeled anti-CD33 in refractory AML (partial) remission could be achieved in 5/18 patients. Randomized trials to evaluate the therapeutic efficacy of RIT in the context of stem cell transplantation have been initiated and the results are keenly anticipated. PMID- 16247434 TI - Management of chronic myeloid leukaemia in relapse following donor lymphocyte infusion induced remission: a retrospective study of the Clinical Trials Committee of the British Society of Blood & Marrow Transplantation (BSBMT). AB - Donor lymphocyte infusion (DLI) can restore remission in a high percentage of patients with chronic myeloid leukaemia (CML) who relapse after allogeneic stem cell transplant (SCT). Subsequent relapses after a DLI-induced remission do occur and the optimal management of these patients is not defined. A retrospective study of the practice of UK transplant centres was conducted. In all, 13 patients from seven centres were identified: all were treated for relapse post allogeneic SCT with DLI and achieved either a complete cytogenetic (n=5) or molecular (n=8) remission. All patients subsequently had a second relapse, at molecular (n=7), cytogenetic (n=4) and haematological (n=2) levels. Further DLI was used in the treatment of 11 patients, imatinib mesylate in three and chemotherapy in two. The two patients with haematological relapse died of blastic disease. The remaining 11 patients achieved either a complete cytogenetic (n=2) or molecular (n=9) remission. Nine patients remain in molecular remission at a median follow-up of 29 months, seven of whom had received DLI alone as treatment for second relapse, one DLI plus imatinib and one imatinib alone. Toxicity following DLI for second relapse was low. Longer follow-up will be required to see if these second DLI induced remissions will be durable. PMID- 16247433 TI - Protective role of interleukin-10 promoter gene polymorphism in the pathogenesis of invasive pulmonary aspergillosis after allogeneic stem cell transplantation. AB - The current study attempted to evaluate the association between the IL-10 promoter gene single nucleotide polymorphism (SNP) and invasive pulmonary aspergillosis (IPA) after allogeneic stem cell transplantation (SCT) in 105 patients. Three single-nucleotide polymorphisms were investigated in the proximal region of the IL-10 promoter gene (-1082/-819/-592). Two haplotypes (1082*A/819*T/592*A [ATA] and 1082*A/819*C/592*C [ACC]) were found in the current study. The overall incidence of IPA was estimated as 14.1+/-4.5% with a median onset at 186 days post-transplant (62 approximately 405 days). An increased occurrence of IPA was noted dependent on the IL-10 haplotype (0% vs 11.5+/-6.4% vs 19.7+/-7.7% for ACC/ACC vs ATA/ACC vs ATA/ATA haplotype, P=0.0307 when comparing ACC with non-ACC haplotype). In a multivariate survival analysis using Cox's proportional hazard model, the IL-10 promoter gene SNPs were identified as an independent predictive factor for the development of IPA (P=0.012, hazard ratio (HR) 9.3), along with an histocompatibility leukocyte antigen (HLA) identical donor (P=0.005, HR 16.3), the CD34+ cell dose transplanted (P=0.004, HR 26.5), and time-dependent chronic graft-versus-host disease (GVHD; P=0.049, HR 16.0). The IL-10 ACC haplotype was found to have an apparent protective role in the development of IPA after allogeneic transplantation, regardless of HLA disparity or chronic GVHD. PMID- 16247435 TI - Graft clonogenicity and intensity of pre-treatment: factors affecting outcome of autologous peripheral hematopoietic cell transplantation in patients with acute myeloid leukemia in first remission. AB - A total of 22 patients with acute myeloid leukemia (AML) in first complete remission receiving autologous blood stem cell transplantation (ABSCT) were investigated in order to determine factors affecting outcome. All but two patients had a normal karyotype and received the same high-dose chemotherapy followed by G-CSF-mobilized peripheral blood stem cells after the second (n=5) or third (n=17) course of induction and post-remission chemotherapy, respectively. With a median follow-up of 30 months, the median disease-free survival is 24.1 months. Univariate analysis showed that three chemotherapy cycles before ABSCT were associated with a significant better disease-free survival (P=0.0018) and overall survival (P=0.0033), whereas the presence of an FLT3-mutation (n=6) showed no impact. The number of megakaryocytic progenitors (CFU-MK) infused tended to correlate with primary platelet engraftment (P=0.07) and were predictive for neutrophil (P=0.011) and platelet counts (P=0.009) 180 days after transplantation. Patients receiving a higher amount of CFU-MK had a better event free survival (P=0.02). Our data suggest that the content of CFU-MK within the graft predicts the quality of hematological recovery and long-term disease control. Additionally, a minimum of three chemotherapy cycles before ABSCT seems to be associated with an improved outcome. PMID- 16247436 TI - Very acute cardiac toxicity during BEAC chemotherapy in non-Hodgkin's lymphoma patients undergoing autologous stem cell transplantation. AB - Cardiotoxicity is potentially the most threatening nonhaematological side effect of high-dose CY. We prospectively evaluated the very acute cardiac effects of high-dose CY in 17 adult non-Hodgkin's lymphoma (NHL) patients receiving CY 1500 mg/m2/day as a part of BEAC high-dose therapy (HDT). Magnetic resonance imaging (MRI) and plasma natriuretic peptide (NT-proBNP, NT-proANP) measurements were performed prior to HDT (d-7) and just after completing HDT (d-2). After the high dose CY left atrial end-systolic area increased from 15.2+/-1.2 to 18.5+/-1.4 cm2 (P=0.001), left ventricular end-diastolic volume from 136.1+/-12.3 to 156.6+/ 11.1 cm3 (P=0.04) and left ventricular end-systolic volume from 67.4+/-7.8 to 75.3+/-7.1 cm3 (P=0.018). However, no significant change in left ventricular ejection fraction (LVEF) was observed. At the same time, plasma levels of NT proBNP increased from 134.9+/-53.3 to 547.1+/-168.4 pmol/l (P=0.003) and NT proANP from 481.1+/-105.5 to 1056.6+/-193.1 pmol/l (P=0.001), respectively. To conclude, high-dose CY results in very acute cardiac toxicity characterised by enlargement of the heart chambers in NHL patients previously treated with anthracyclines. This toxicity can be detected with increased concentrations of circulating natriuretic peptides but not with LVEF measurement. PMID- 16247438 TI - Acute myeloid leukemia with near-triploid karyotype and extramedullary involvement of mediastinum. PMID- 16247437 TI - Demonstration of reversed flow in segmental branches of the portal vein with hand held color Doppler ultrasonography after hematopoietic stem cell transplantation. AB - Hepatic veno-occlusive disease (VOD) is a severe complication of hematopoietic stem cell transplantation (SCT). When monitored with hand-held color Doppler ultrasonography during day -7 to +35 around SCT, reversed blood flow in the segmental branches of the portal vein was detected in nine of 56 patients who had undergone SCT. Three of nine patients had clinical evidence of VOD, but six patients did not fulfill the criteria for diagnosis of VOD initially. Two patients progressed to clinical VOD at a later date and the reversed portal flow disappeared with or without treatment for VOD in the other four patients. Monitoring for reversed portal flow with color Doppler ultrasonography may be a useful tool for the early diagnosis of VOD, and may improve prognosis by allowing early initiation of treatment. PMID- 16247439 TI - Outlook: finding improved medicines: the role of academic-industrial collaboration. AB - This paper reviews models of academic-pharmaceutical industry collaboration and debates the value of such partnerships so that those contemplating an alliance can reflect a priori on the purpose, nature and process that will provide a constructive outcome. The scope is confined to the biomedical discipline, because collaborations in other fields, such as physics and engineering, have not suffered as a result of the concerns associated with biomedicine. PMID- 16247440 TI - Outlook: the profit problem in antibiotic R&D. AB - Economists, a biomedical researcher and a business executive have formulated three contrasting proposals to address the shortfall in antibiotic R&D. Their proposals, which emphasize advance purchase contracts, not-for-profit research, and tax incentives, respectively, share some features with provisions in bills pending before the US Congress that could potentially reshape the R&D landscape for this essential class of drugs. PMID- 16247441 TI - Phosphatidylinositol 4-phosphatase type II is an erythropoietin-responsive gene. AB - The erythroleukemia developed by spi-1/PU.1 transgenic mice is a multistep process. At disease onset, preleukemic cells are arrested in differentiation at the proerythroblast stage (HS1 stage) and their survival and growth are under the tight control of erythropoietin (Epo). During disease progression, malignant proerythroblasts characterized by Epo autonomous growth and in vivo tumorigenicity can be isolated (HS2 stage). During analysis of transcriptional profiling representive of discrete stages of leukemic progression, we found that the phosphatidylinositol 4-phosphatase type II gene was turned off in malignant cells. PI-4-phosphatase II is an enzyme that hydrolyses the 4-phosphate position of phosphatidylinositol-3-4-bisphosphate (PtdIns(3,4)P(2)) to form PtdIns(3)P. Using malignant cells engineered to stably express PI-4-phosphatase II, we showed that PI-4-phosphatase II reduced Akt activation level. Moreover, stimulation of malignant cells with Epo-induced PI-4-phosphatase II transcription pointing this gene as an Epo-responsive gene. This study provides first insight for a physiological role of PI-4-phosphatase II in the proerythroblast by controlling Epo responsiveness through a negative regulation of the PI3K/Akt pathway. PMID- 16247442 TI - p53 promotes adenoviral replication and increases late viral gene expression. AB - The tumor suppressor protein, p53, plays a critical role in viro-oncology. However, the role of p53 in adenoviral replication is still poorly understood. In this paper, we have explored further the effect of p53 on adenoviral replicative lysis. Using well-characterized cells expressing a functional p53 (A549, K1neo, RKO) and isogenic derivatives that do not (K1scx, RKOp53.13), we show that virus replication, late virus protein expression and both wtAd5 and ONYX-015 virus induced cell death are impaired in cells deficient in functional p53. Conversely, by transfecting p53 into these and other cells (IIICF/c, HeLa), we increase late virus protein expression and virus yield. We also show, using reporter assays in IIICF/c, HeLa and K1scx cells, that p53 can cooperate with E1a to enhance transcription from the major late promoter of the virus. Late viral protein production is enhanced by exogenous p53. Taken together, our data suggest that functional p53 can promote the adenovirus (Ad) lytic cycle. These results have implications for the use of Ad mutants that are defective in p53 degradation, such as ONYX-015, as agents for the treatment of cancers. PMID- 16247443 TI - Cleavage of epidermal growth factor receptor by caspase during apoptosis is independent of its internalization. AB - Epidermal growth factor receptor (EGFR) plays a critical role in cell proliferation, differentiation, and transformation. EGFR downregulation attenuates its signaling intensity and duration to maintain cellular homeostasis. Here, we report that during apoptosis EGFR is cleaved by activated caspase-3 or related proteases at its C-terminus domain. EGFR downregulation by activation of caspases is neither stimulus- nor cell type-specific. EGFR internalization during apoptosis required dynamin and cholesterol since dominant-negative dynamin (K44A) or cholesterol depletion by methyl-beta-cyclodextrin prevented EGFR internalization. However, EGFR downregulation did not require its internalization. The EGFR cleavage fragment was detected in the membrane blebs in addition to the cell pellets. Mutations at the consensus sequence (DXXD) at the C terminus domain revealed that DVVD1012 and to a lesser extent DNPD1172 may be target sites for active recombinant caspase-3 in vitro and activated caspase-3 or related proteases in vivo. We have detected the N-terminus and C-terminus fragments in vitro and in vivo. A cleavage-deficient EGFR mutant delayed apoptosis process. We conclude that the evolutionarily conserved C-terminus domain of EGFR is the target of caspases and subjected to degradation during apoptosis to shut down its signaling. PMID- 16247444 TI - A mouse model for tumor progression of lung cancer in ras and p53 transgenic mice. AB - Although ras and p53 are the most commonly found oncogene and tumor suppressor gene, respectively, in human cancers, their collective roles in tumor progression have yet to be defined in animal models. Here, we demonstrated the synergistic effect between ras and p53 in promoting tumor progression during lung tumorigenesis using bitransgenic mice. Mice with a heterozygous knockout of K-ras (K-ras(wt/ko)) were mated to p53 transgenic mice (p53(val135/wt)) in lung tumorigenesis (K-ras(wt/ko) x p53(val135/wt)). F(1) mice exhibited a significant increase in lung tumor load (tumor multiplicity x tumor volume) when compared to those seen in either K-ras(wt/ko) mice or p53(val135/wt) mice alone. Furthermore, over 50% of the lung tumors were lung adenocarcinomas in bitransgenic mice compared to only 3% in wild-type mice. Alterations of ras and p53 appear to promote the development of lung adenocarcinomas. These results provide the in vivo experimental evidence of synergistic interactions of ras and p53 in lung tumor progression. PMID- 16247445 TI - ELA2 is regulated by hematopoietic transcription factors, but not repressed by AML1-ETO. AB - A 117 bp fragment of the human ELA2 promoter has been characterized that can act as a minimal promoter for the expression of neutrophil elastase. Chromatin immunoprecipitation and siRNAs revealed that expression of ELA2 is regulated by the acute myeloid human leukemia 1 protein (AML1), C/EBPalpha, PU.1 and c-Myb transcription factors. ELA2 has also been investigated as a possible target of the leukemic fusion protein AML1-ETO resulting from the t(8;21) chromosomal translocation. AML1-ETO, like AML1, binds the ELA2 promoter in the myeloid cell lines Kasumi-1 and U937, but unexpectedly fails to significantly alter expression of ELA2. Although AML1-ETO downregulates the expression of C/EBPalpha, changes in C/EBPalpha expression do not correlate with changes in the expression of ELA2. Our observations indicate that AML1-ETO may not be a constitutive repressor of gene expression in every case in which it can associate with DNA, either on its own or in conjunction with C/EBPalpha. Since neither ETO nor AML1-ETO are typically expressed in hematopoietic progenitors, we hypothesize that it is the interactions between AML1-ETO and regulatory cofactors in disease-state cells that alter gene expression programs during hematopoiesis. These protein-protein interactions may not require simultaneous DNA binding by AML1-ETO for the deleterious effects of the fusion protein to be realized. PMID- 16247446 TI - Prevention of lung cancer progression by bexarotene in mouse models. AB - Bexarotene (Targretin), is a synthetic high-affinity RXR receptor agonist with limited affinity for RAR receptors. Bexarotene has shown efficacy in a phase I/II trial of non-small-cell lung cancers. However, the chemopreventive efficacy of bexarotene has not been determined in mouse lung cancer models. In this study, we have investigated the ability of bexarotene to inhibit lung tumor progression in the mutant A/J mouse models with genetic alterations in p53 or K-ras, two of the most commonly altered genes in human lung tumorigenesis. Mice were administered vinyl carbamate (VC), a carcinogen, by a single intraperitoneal injection (i.p.) at 6 weeks of age. Bexarotene was given by gavage starting at 16 weeks after VC and was continued for 12 weeks. Although all mice developed lung tumors, only 7% of lung tumors were adenocarcinomas in wild-type mice, whereas 22 and 26% of lung tumors were adenocarcinomas in p53 transgenic or K-ras heterozygous deficient mice. Bexarotene inhibited both tumor multiplicity and tumor volume in mice of all three genotypes. Furthermore, bexarotene reduced the progression of adenoma to adenocarcinoma by approximately 50% in both p53(wt/wt)K-ras(ko/wt) and p53(wt/wt)K-ras(wt/wt) mice. Thus, bexarotene appears to be an effective preventive agent against lung tumor growth and progression. PMID- 16247448 TI - The fate of irradiated tumor cells. AB - Radiotherapy remains a key component of modern multimodal anticancer treatment approaches. Although most clinically relevant improvements in radiation oncology are based on optimized radiation technology, it now becomes obvious that biology based approaches will further boost the efficacy of modern radiation oncology. Of relevance to this is the question of the ultimate fate of an irradiated cell and the mechanistic basis of the induced cell death or inactivation observed. Research bearing on this question is reported in this issue of Oncogene by Wendt et al. (2005), who show that the levels of p21CIP/WAF1 play a key role in determining whether an irradiated tumour cell is arrested in the G2 phase of the cell cycle, rather than apoptosing. The demonstration that caffeine treatment can abrogate this G2 arrest, and that the cells go on to die, has implications for overcoming treatment resistance imposed by radiation-induced upregulation of p21CIP/WAF1. PMID- 16247447 TI - Identification of novel genomic markers related to progression to glioblastoma through genomic profiling of 25 primary glioma cell lines. AB - Identification of genetic copy number changes in glial tumors is of importance in the context of improved/refined diagnostic, prognostic procedures and therapeutic decision-making. In order to detect recurrent genomic copy number changes that might play a role in glioma pathogenesis and/or progression, we characterized 25 primary glioma cell lines including 15 non glioblastoma (non GBM) (I-III WHO grade) and 10 GBM (IV WHO grade), by array comparative genomic hybridization, using a DNA microarray comprising approx. 3500 BACs covering the entire genome with a 1 Mb resolution and additional 800 BACs covering chromosome 19 at tiling path resolution. Combined evaluation by single clone and whole chromosome analysis plus 'moving average (MA) approach' enabled us to confirm most of the genetic abnormalities previously identified to be associated with glioma progression, including +1q32, +7, -10, -22q, PTEN and p16 loss, and to disclose new small genomic regions, some correlating with grade malignancy. Grade I-III gliomas exclusively showed losses at 3p26 (53%), 4q13-21 (33%) and 7p15-p21 (26%), whereas only GBMs exhibited 4p16.1 losses (40%). Other recurrent imbalances, such as losses at 4p15, 5q22-q23, 6p23-25, 12p13 and gains at 11p11 q13, were shared by different glioma grades. Three intervals with peak of loss could be further refined for chromosome 10 by our MA approach. Data analysis of full-coverage chromosome 19 highlighted two main regions of copy number gain, never described before in gliomas, at 19p13.11 and 19q13.13-13.2. The well-known 19q13.3 loss of heterozygosity area in gliomas was not frequently affected in our cell lines. Genomic hotspot detection facilitated the identification of small intervals resulting in positional candidate genes such as PRDM2 (1p36.21), LRP1B (2q22.3), ADARB2 (10p15.3), BCCIP (10q26.2) and ING1 (13q34) for losses and ECT2 (3q26.3), MDK, DDB2, IG20 (11p11.2) for gains. These data increase our current knowledge about cryptic genetic changes in gliomas and may facilitate the further identification of novel genetic elements, which may provide us with molecular tools for the improved diagnostics and therapeutic decision-making in these tumors. PMID- 16247449 TI - RhoB facilitates c-Myc turnover by supporting efficient nuclear accumulation of GSK-3. AB - The small GTPase RhoB suppresses cancer in part by limiting cell proliferation. However, the mechanisms it uses to achieve this are poorly understood. Recent studies link RhoB to trafficking of Akt, which through its regulation of glycogen synthase kinase-3 (GSK-3) has an important role in controlling the stability of the c-Myc oncoprotein. c-Myc stabilization may be a root feature of human tumorigenesis as it phenocopies an essential contribution of SV40 small T antigen in human cell transformation. In this study we show that RhoB directs efficient turnover of c-Myc in established or transformed mouse fibroblasts and that the attenuation of RhoB which occurs commonly in human cancer is a sufficient cause to elevate c-Myc levels. Increased levels of c-Myc elicited by RhoB deletion increased the proliferation of nullizygous cells, whereas restoring RhoB in null cells decreased the stability of c-Myc and restrained cell proliferation. Mechanistic analyses indicated that RhoB facilitated nuclear accumulation of GSK 3 and GSK-3-mediated phosphorylation of c-Myc T58, the critical site for ubiquitination and degradation of c-Myc. RhoB deletion restricted nuclear localization of GSK-3, reduced T58 phosphorylation, and stabilized c-Myc. These effects were not associated with changes in phosphorylation or localization of Akt, however, differences were observed in phosphorylation and localization of the GSK-3 regulatory Akt-related kinase, serum- and glucocorticoid-inducible protein kinase (SGK). The ability of RhoB to support GSK-3-dependent turnover of c-Myc offers a mechanism by which RhoB acts to limit the proliferation of neoplastically transformed cells. PMID- 16247451 TI - RNAi-based screening of the human kinome identifies Akt-cooperating kinases: a new approach to designing efficacious multitargeted kinase inhibitors. AB - Tumors comprise genetically heterogeneous cell populations, whose growth and survival depend on multiple signaling pathways. This has spurred the development of multitargeted therapies, including small molecules that can inhibit multiple kinases. A major challenge in designing such molecules is to determine which kinases to inhibit in each cancer to maximize efficacy and therapeutic index. We describe an approach to this problem implementing RNA interference technology. In order to identify Akt-cooperating kinases, we screened a library of kinase directed small interfering RNAs (siRNAs) for enhanced cancer cell killing in the presence of Akt inhibitor A-443654. siRNAs targeting casein kinase I gamma 3 (CSNK1G3) or the inositol polyphosphate multikinase (IPMK) significantly enhanced A-443654-mediated cell killing, and caused decreases in Akt Ser-473 and ribosomal protein S6 phosphorylation. Small molecules targeting CSNK1G3 and/or IPMK in addition to Akt may thus exhibit increased efficacy and have the potential for improved therapeutic index. PMID- 16247450 TI - Cell context reveals a dual role for Maf in oncogenesis. AB - Maf b-Zip transcription factors are involved in both terminal differentiation and oncogenesis. To investigate this apparent contradiction, we used two different primary cell types and performed an extensive analysis of transformation parameters induced by Maf proteins. We show that MafA and c-Maf are potent oncogenes in chicken embryo fibroblasts, while MafB appears weaker. We also provide the first evidence that MafA can confer growth factor independence and promote cell division at low density. Moreover, using MafA as a model, we identified several parameters that are critical for Maf transforming activities. Indeed, MafA ability to induce anchorage-independent cell growth was sensitive to culture conditions. In addition, the transforming activity of MafA was dependent on its phosphorylation state, since mutation on Ser65 impaired its ability to induce growth at low density and anchorage-independent growth. We next examined transforming activity of large Maf proteins in embryonic neuroretina cells, where they are known to induce differentiation. Unlike v-Jun, MafA, MafB and c-Maf did not show oncogenic activity in these cells. Moreover, they counteracted transformation induced by constitutive activation of the Ras/Raf/MEK pathway. Taken together, our results show that Maf proteins could display antagonistic functions in oncogenesis depending on the cellular context, and support a dual role for Maf as both oncogenes and tumor suppressor-like proteins. PMID- 16247452 TI - Syndecan-1 expression by stromal fibroblasts promotes breast carcinoma growth in vivo and stimulates tumor angiogenesis. AB - The induction of the cell surface heparan sulfate proteoglycan syndecan-1 (Sdc1) in stromal fibroblasts is observed in more than 70% of human breast carcinomas. Using a coculture model, we have recently shown that stromal cell-derived Sdc1 stimulates carcinoma cell proliferation in vitro, and that this activity requires Sdc1 glycanation. In the present study, we investigated the effect of stromal cell Sdc1 on breast carcinoma growth in vivo. MDA-MB-231 human breast carcinoma cells were inoculated into the flanks of athymic nude mice either alone, or as mixed suspensions with Sdc1-transfected or mock-transfected 3T3 mouse fibroblasts. The mixed tumors showed an intimate association between carcinoma cells and stromal fibroblasts and histologically closely resembled poorly differentiated human breast carcinomas. The presence of fibroblasts led to significantly accelerated tumor growth, which was further augmented (88% increase) by forced expression of stromal Sdc1. The hyperemic macroscopic appearance of tumors containing Sdc1-positive stromal cells contrasted with pale tumors developing in the presence of mock-transfected fibroblasts, which prompted us to examine tumor microvessels. Stromal Sdc1 expression was associated with a significantly elevated microvessel density (36% increase) and a larger vessel area (153% increase). To evaluate the relevance of this finding in human breast cancer, the relationship between stromal Sdc1 and tumor vascularity was also examined in a tissue array containing 207 human breast carcinoma samples. Similar to the xenografts, stromal Sdc1 expression correlated with both vessel density (P=0.013) and total vessel area (P=0.0026). In conclusion, stromal fibroblast derived Sdc1 stimulates breast carcinoma growth and angiogenesis in vivo. PMID- 16247453 TI - DNA copy number gains in head and neck squamous cell carcinoma. AB - Gene amplification, a common mechanism for oncogene activation in cancer, has been used as a tag for the identification of novel oncogenes. DNA amplification is frequently observed in head and neck squamous cell carcinoma (HNSCC) and potential oncogenes have already been reported. We applied restriction landmark genome scanning (RLGS) to study gene amplifications and low-level copy number changes in HNSCC in order to locate previously uncharacterized regions with copy number gains in primary tumor samples. A total of 63 enhanced RLGS fragments, indicative of DNA copy number changes, including gains of single alleles, were scored. Enhanced sequences were identified from 33 different chromosomal regions including those previously reported (e.g. 3q26.3 and 11q13.3) as well as novel regions (e.g. 3q29, 8q13.1, 8q22.3, 9q32, 10q24.32, 14q32.32, 17q25.1 and 20q13.33). Furthermore, our data suggest that amplicons 11q13.3 and 3q26.3-q29 may be divided into possibly two and three independent amplicons, respectively, an observation supported by published microarray expression data. PMID- 16247454 TI - beta1-integrin-mediated signaling essentially contributes to cell survival after radiation-induced genotoxic injury. AB - Integrin-mediated adhesion to extracellular matrix proteins confers resistance to radiation- or drug-induced genotoxic injury. To analyse the underlying mechanisms specific for beta1-integrins, wild-type beta1A-integrin-expressing GD25beta1A cells were compared to GD25beta1B cells, which express signaling-incompetent beta1B variants. Cells grown on fibronectin, collagen-III, beta1-integrin-IgG or poly-l-lysine were exposed to 0-6 Gy X-rays in presence or depletion of growth factors and phosphatidylinositol-3 kinase (PI3K) inhibitors (LY294002, wortmannin). In order to test the relevance of these findings in tumor cells, human A-172 glioma cells were examined under the same conditions after siRNA mediated silencing of beta1-integrins. We found that beta1A-integrin-mediated adhesion to fibronectin, collagen-III or beta1-IgG was essential for cell survival after radiation-induced genotoxic injury. Mediated by PI3K, pro-survival beta1A-integrin/Akt signaling was critically involved in this process. Additionally, the beta1-integrin downstream targets p130Cas and paxillin-impaired survival-regulating PI3K-dependent JNK. In A-172 glioma cells, beta1-integrin knockdown and PI3K inhibition confirmed the central role of beta1-integrins in Akt- and p130Cas/paxillin-mediated prosurvival signaling. These findings suggest beta1-integrins as critical regulators of cell survival after radiation-induced genotoxic injury. Elucidation of the molecular circuitry of prosurvival beta1 integrin-mediated signaling in tumor cells may promote the development of innovative molecular-targeted therapeutic antitumor strategies. PMID- 16247455 TI - The JAK2 V617F mutation in de novo acute myelogenous leukemias. AB - A missense somatic mutation in JAK2 gene (JAK2 V617F) has recently been reported in chronic myeloproliferative disorders, including polycythemia vera, essential thrombocythemia and myelofibrosis with myeloid metaplasia, strongly suggesting its role in the pathogenesis of myeloid disorders. As activation of JAK2 signaling is occurred in other malignancies as well, we have analysed 558 tissues from common human cancers, including colon, breast and lung carcinomas, and 143 acute adulthood leukemias by polymerase chain reaction -- single strand conformation polymorphism analysis. We found three JAK2 mutations in the 113 acute myelogenous leukemias (AMLs) (2.7%), but none in other cancers. The mutations consisted of two V617F mutations and one K607N mutation. None of the AML patients with the JAK2 V617F mutation had a history of previous hematologic disorders. This is the first report on the JAK2 gene mutation in AML, and the data indicated that the JAK2 gene mutation may not only contribute to the development of chronic myeloid disorders, but also to some AMLs. PMID- 16247456 TI - Defect in serine 46 phosphorylation of p53 contributes to acquisition of p53 resistance in oral squamous cell carcinoma cells. AB - To investigate whether dysregulation of p53 phosphorylation confers tumor resistance to p53, we analysed the effects of wild-type p53 on oral squamous cell carcinoma (SCC) cell lines carrying various mutations of p53. Introduction of exogenous p53 neither induced apoptosis nor suppressed colony formation in HSC-3 cells lacking any detectable p53 and HSC-4 cells expressing mutant p53R248Q protein. Consistently, exogenous p53 did not induce proapoptotic p53-target genes in these p53-resistant cells. We found that phosphorylation of exogenous p53 on serine 46 (Ser46) was severely impaired in HSC-3 but not HSC-4 cells. A mutant mimicking Ser46-phosphorylation (p53S46D) enhanced proapoptotic Noxa promoter activity, and overcame the resistance to p53-mediated apoptosis and growth suppression in HSC-3 cells. Conversely, a mutant defective for Ser46 phosphorylation (p53S46A) failed to suppress the growth of p53-sensitive HSC-2 cells. In contrast to HSC-3 cells, p53S46D had no effect on HSC-4 cells, and inhibition of endogenous p53R248Q by siRNA restored p53-mediated apoptosis in HSC 4 cells, indicating a dominant-negative effect of p53R248Q protein on wild-type p53 function. These results demonstrate that the defect in Ser46 phosphorylation accounts for the p53 resistance of HSC-3 cells, and provide evidence for a mechanism underlying the acquisition of p53 resistance in oral SCC. PMID- 16247457 TI - Molecular determinants of Akt-induced keratinocyte transformation. AB - The PI3K/PTEN/Akt signaling pathway has emerged in recent years as a main player in human cancers, increasing proliferation and decreasing apoptosis of transformed cells, and thus becoming a potential target for therapeutic intervention. Our previous data have demonstrated that Akt-mediated signaling is of a key relevance in the mouse skin carcinogenesis system, one of the best-known models of experimental carcinogenesis. Here, we investigated the involvement of several pathways as mediators of Akt-induced increased proliferation and tumorigenesis in keratinocytes. Tumors produced by subcutaneous injection of Akt transformed keratinocytes showed increased Foxo3a phosphorylation, but no major alterations in p21(Cip1/WAF1), p27(Kip1) or mdm2 expression and/or localization. In contrast, we found increased expression and nuclear localization of DeltaNp63, beta-catenin and Lef1. Concomitantly, we also found increased expression of c-myc and CycD1, targets of the beta-catenin/Tcf pathway. Such increase is associated with increased phosphorylation and stabilization of c-myc protein as well as increased translation of c-myc and CycD1 due to mTOR activation. Using immunohistochemistry approaches in samples of oral dysplasias and human head and neck squamous cell carcinomas, we confirmed that increased Akt activation significantly correlates with increased DeltaNp63 and CycD expression, c-myc phosphorylation and nuclear accumulation of beta-catenin. Collectively, these results demonstrate that Akt is able to transform keratinocytes by specific mechanisms involving transcriptional and post-transcriptional processes. PMID- 16247458 TI - Functional epigenetics identifies a protocadherin PCDH10 as a candidate tumor suppressor for nasopharyngeal, esophageal and multiple other carcinomas with frequent methylation. AB - Protocadherins constitute the largest subgroup in the cadherin superfamily of cell adhesion molecules. Their major functions are poorly understood, although some are implicated in nervous system development. As tumor-specific promoter methylation is a marker for tumor suppressor genes (TSG), we searched for epigenetically inactivated TSGs using methylation-subtraction combined with pharmacologic demethylation, and identified the PCDH10 CpG island as a methylated sequence in nasopharyngeal carcinoma (NPC). PCDH10 is broadly expressed in all normal adult and fetal tissues including the epithelia, though at different levels. It resides at 4q28.3--a region with hemizygous deletion detected by array CGH in NPC cell lines; however, PCDH10 itself is not located within the deletion. In contrast, its transcriptional silencing and promoter methylation were frequently detected in multiple carcinoma cell lines in a biallelic way, including 12/12 nasopharyngeal, 13/16 esophageal, 3/4 breast, 5/5 colorectal, 3/4 cervical, 2/5 lung and 2/8 hepatocellular carcinoma cell lines, but not in any immortalized normal epithelial cell line. Aberrant methylation was further frequently detected in multiple primary carcinomas (82% in NPC, 42-51% for other carcinomas), but not normal tissues. The transcriptional silencing of PCDH10 could be reversed by pharmacologic demethylation with 5-aza-2'-deoxycytidine or genetic demethylation with double knockout of DNMT1 and DNMT3B, indicating a direct epigenetic mechanism. Ectopic expression of PCDH10 strongly suppressed tumor cell growth, migration, invasion and colony formation. Although the epigenetic and genetic disruptions of several classical cadherins as TSGs have been well documented in tumors, this is the first report that a widely expressed protocadherin can also function as a TSG that is frequently inactivated epigenetically in multiple carcinomas. PMID- 16247459 TI - Merlin facilitates ubiquitination and degradation of transactivation-responsive RNA-binding protein. AB - The Nf2 tumor suppressor codes for merlin, a protein whose function is largely unknown. We have previously demonstrated a novel interaction between merlin and TRBP, which inhibits the oncogenic activity of TRBP. In spite of the significance of their functional interaction, its molecular mechanism still remains to be elucidated. In this report, we investigated how merlin inhibits the oncogenic activity of TRBP in association with cell growth conditions. In the human embryonic kidney 293 cell line, the level of endogenous merlin increased, whereas that of endogenous TRBP significantly decreased along with the increase in cell confluence. We demonstrated that the carboxyl-terminal region of TRBP was responsible for this phenomenon using stable cell lines expressing deletion mutants of TRBP. The overexpression of merlin decreased the protein level of TRBP, and the ubiquitin-like subdomain of merlin's FERM domain was important for this activity. We also demonstrated that TRBP is ubiquitinylated and the ubiquitinylated forms of TRBP are accumulated by ectopically expressed merlin or cell confluence in the presence of MG132, a proteasome inhibitor. Furthermore, we showed that the regulation of TRBP in response to cell confluence was abolished upon knockdown of merlin expression by specific small interfering RNA. Finally, we showed that ectopically expressed merlin restored cell-cell contact inhibition in cells stably expressing TRBP but not in TRBPDeltac. These results suggest that merlin is involved in the regulation of TRBP protein level by facilitating its ubiquitination in response to such cues as cell-cell contacts. PMID- 16247461 TI - IGF2 is critical for tumorigenesis by synovial sarcoma oncoprotein SYT-SSX1. AB - Synovial sarcoma is an aggressive soft tissue tumor characterized by a specific chromosomal translocation between chromosome 18 and X. This translocation can generate a fusion transcript encoding SYT-SSX1, a transforming oncoprotein. We present evidence that SYT-SSX1 induces insulin-like growth factor II expression in fibroblast cells. SYT-SSX2, a fusion also frequently found in synovial sarcoma, is necessary for maintaining Igf2 expression in the synovial sarcoma cell line, and the increased IGF2 synthesis protects cells from anoikis and is required for tumor formation in vivo. We also found a loss of imprinting (LOI) for Igf2 in a limited number of primary synovial sarcomas despite demethylation of CpG dinucleotides critical for maintaining imprinting. These findings suggest that inhibition of the IGF2/IGF1-R signaling pathway may represent a significant therapeutic modality for treating synovial sarcoma. PMID- 16247460 TI - Expression of constitutively nuclear cyclin D1 in murine lymphocytes induces B cell lymphoma. AB - Mantle cell lymphoma (MCL) is a B-cell lymphoma characterized by overexpression of cyclin D1 due to the t(11;14) chromosomal translocation. While expression of cyclin D1 correlates with MCL development, expression of wild-type (WT) cyclin D1 transgene in murine lymphocytes is unable to drive B-cell lymphoma. As cyclin D1 mutants that are refractory to nuclear export display heighten oncogenicity in vitro compared with WT D1, we generated mice expressing FLAG-D1/T286A, a constitutively nuclear mutant, under the control of the immunoglobulin enhancer, Emu. D1/T286A transgenic mice universally develop a mature B-cell lymphoma. Expression of D1/T286A in B lymphocytes results in S phase entry in resting lymphocytes and increased apoptosis in spleens of young premalignant mice. Lymphoma onset correlates with perturbations in p53/MDM2/p19Arf expression and with BcL-2 overexpression suggesting that alterations in one or both of these pathways may contribute to lymphoma development. Our results describe a cyclin D1 driven model of B-cell lymphomagenesis and provide evidence that nuclear retention of cyclin D1 is oncogenic in vivo. PMID- 16247463 TI - Clusterin plays an important role in hepatocellular carcinoma metastasis. AB - To identify genes associated with tumor metastasis in hepatocellular carcinoma (HCC), gene expression profiles between a pair of primary HCC (H2-P) and their matched metastatic HCC (H2-M) were compared. Overexpression of clusterin (CLU) was found in H2-M cells. To determine the roles CLU played in HCC metastasis, CLU was transfected into H2-P cells. Overexpression of CLU in H2-P cells increased cell migration by twofold in vitro and formation of metastatic tumor nodules in liver by eightfold in vivo. To evaluate the correlation of CLU expression with HCC metastasis, the expression levels of CLU in HCCs were investigated using a tissue microarray (TMA) containing 104 pairs of primary HCCs and their matched metastases. The frequency of CLU overexpression increased significantly in metastatic HCCs (59.1%) compared with that in primary tumors (32.6%, P<0.001). To gain additional insight into the function of CLU, the expression profile of H2P CLU was compared with vector-transfected H2-P cells by cDNA microarray. A total of 35 upregulated and 14 downregulated genes were detected in H2P-CLU. One of the upregulated genes known as YKL-40, which is implicated in matrix-remodeling and metastasis, was further studied using TMA. A significant correlation (P<0.001) between the expression levels of YKL-40 and CLU was observed, implying that the CLU-YKL-40 pathway may play an important role in HCC metastasis. PMID- 16247462 TI - Adenoviral modification of mouse brain derived endothelial cells, bEnd3, to induce apoptosis by vascular endothelial growth factor. AB - A second generation genetically-engineered cell-based drug delivery system, referred to as apoptotic-induced drug delivery (AIDD), was developed using endothelial cells (ECs) that undergo apoptosis upon binding of vascular endothelial growth factor (VEGF) to a Flk-1:Fas fusion protein (FF). This new AIDD was redesigned using mouse brain derived ECs, bEnd3 cells, and an adenovirus vector in order to enhance and control the expression of FF. The FF was tagged with a HA epitope (FFHA) and designed to be coexpressed with green fluorescence protein (GFP) by the regulation of cytomegalovirus promoters in the adenovirus vector. bEnd3 cells showed favorable coexpression of FFHA and GFP consistent with the multiplicity of infection of the adenovirus. Immunofluorescence analysis demonstrated that FFHA was localized at the plasma membrane, whereas GFP was predominantly located in the cytoplasm of ECs. Cell death was induced by VEGF, but not by platelet derived growth factor or fibroblast growth factor in a dose dependent manner (range 2-20 ng/ml), and revealed caspase-dependent apoptotic profiles. The FFHA expressing bEnd3 cells underwent apoptosis when cocultured with a glioma cell (SF188V+) line able to overexpress VEGF. The combined data indicated that the FFHA adenovirus system can induce apoptotic signaling in ECs in response to VEGF, and thus, is an instrumental modification to the development of AIDD. PMID- 16247464 TI - Downregulation of E-cadherin by hepatitis B virus X antigen in hepatocellullar carcinoma. AB - Hepatitis B virus (HBV)-encoded X antigen (HBxAg) contributes to the development of hepatocellular carcinoma (HCC). A frequent characteristic of HCC is reduced or absent expression of the cell adhesion protein, E-cadherin, although it is not known whether HBxAg plays a role. To address this, the levels of E-cadherin were determined in HBxAg-positive and -negative HepG2 cells in culture, and in tumor and surrounding nontumor liver from a panel of HBV carriers. The results showed an inverse relationship between HBxAg and E-cadherin expression both in tissue culture and in vivo. In HBxAg-positive cells, E-cadherin was suppressed at both the mRNA and protein levels. This was associated with hypermethylation of the E cadherin promoter. Depressed E-cadherin correlated with HBxAg trans-activation function, as did the migration of HepG2 cells in vitro. Decreased expression of E cadherin was also associated with the accumulation of beta-catenin in the cytoplasm and/or nuclei in tissues and cell lines, which is characteristic of activated beta-catenin. Additional work showed that HBxAg-activated beta-catenin. Together, these results suggest that the HBxAg is associated with decreased expression of E-cadherin, accumulation of beta-catenin in the cytoplasm and nucleus, and increased cell migration, which may contribute importantly to hepatocarcinogenesis. PMID- 16247465 TI - AML1 deletion in adult mice causes splenomegaly and lymphomas. AB - AML1 (RUNX1) encodes a DNA-binding subunit of the CBF transcription factor family and is required for the establishment of definitive hematopoiesis. AML1 is one of the most frequently mutated genes associated with human acute leukemia, suggesting that genetic alterations of the gene contribute to leukemogenesis. Here, we report the analysis of mice carrying conditional AML1 knockout alleles that were inactivated using the Cre/loxP system. AML1 was deleted in adult mice by inducing Cre activity to replicate AML1 deletions found in human MDS, familial platelet disorder and rare de novo human AML. At a latency of 2 months after induction, the thymus was reduced in size and frequently populated by immature double negative thymocytes, indicating defective T-lymphocyte maturation, resulting in lymphatic diseases with 50% penetrance, including atypical hyperplasia and thymic lymphoma. Metastatic lymphomas to the liver and the meninges were observed. Mice also developed splenomegaly with an expansion of the myeloid compartment. Increased Howell-Jolly body counts indicated splenic hypofunction. Thrombocytopenia occurred due to immaturity of mini-megakaryocytes in the bone marrow. Together with mild lymphocytopenia in the peripheral blood and increased fractions of immature cells in the bone marrow, AML1 deficient mice display features of a myelodysplastic syndrome, suggesting a preleukemic state. PMID- 16247466 TI - MCM7 amplification and overexpression are associated with prostate cancer progression. AB - The genomic DNA profiles of prostate cancers with aggressive features were compared to the profiles of matched normal DNA to identify genes that are selectively amplified in the cancer cells. One of the identified genes, MCM7, which is a component of the DNA replication licensing complex, has been studied extensively both at the DNA and protein levels in human prostate tissues. Approximately half of the prostate cancer specimens studied showed MCM7 gene amplification, and 60% of the aggressive prostate cancer specimens had increased MCM7 protein expression. Amplification or overexpression of MCM7 was significantly associated with relapse, local invasion and a worse tumor grade. Constitutive expression of MCM7 in a human prostate cancer cell line, DU145, resulted in markedly increased DNA synthesis and cell proliferation compared to vector-only controls, and an increased cell invasion in vitro. Indeed, MCM7 overexpression produced primary tumors 12 times larger than vector-only controls and resulted in a rapid demise of mice bearing those tumors. These studies implicate MCM7, and the DNA replication licensing gene family, in prostate cancer progression, growth and invasion. PMID- 16247467 TI - Neoplastic transformation by the gep oncogene, Galpha12, involves signaling by STAT3. AB - Galpha(12), the alpha-subunit of G12, which has been referred to as the gep oncogene, stimulates mitogenic pathways in different cell types and readily induces neoplastic transformation of fibroblast cell lines. Recently, we have shown that the oncogenic pathway activated by Galpha(12) involves the receptor tyrosine kinase platelet derived growth factor receptor-alpha (PDGFRalpha) and JAK3. In the present study, we demonstrate that the GTPase-deficient activated mutant of Galpha(12) activates signal transducer and activator of transcription 3 (STAT3) via PDGFRalpha as well as JAK3. Here we show that Galpha(12) stimulates the phosphorylation of STAT3 at both Tyrosine-705 and Serine-727 residues. Studies to delineate the mechanism by which Galpha(12) stimulates STAT3 have indicated that the Tyrosine-705-phosphorylation of STAT3 involves the tyrosine kinases, Janus Kinase-3 as well as Src kinase, whereas the Serine-727 phosphorylation of STAT3 occurs via the receptor tyrosine kinase, PDGFRalpha and phosphatidylinositol 3-OH kinase pathway. Our results also indicate that the coexpression of the dominant negative, DNA binding mutant of STAT3 (STAT3DB) inhibits the foci formation as well as anchorage-independent growth of Galpha(12)QL-transfectants, thereby establishing the critical role of STAT3 in Galpha(12)QL-mediated neoplastic cell growth. The results presented here demonstrate, for the first time, the ability of Galpha(12) to recruit multiple receptor-, nonreceptor-, and Ser/Thr kinases to stimulate STAT3-signaling to promote neoplastic transformation. PMID- 16247469 TI - Hypoxic regulation of stability of connective tissue growth factor/CCN2 mRNA by 3'-untranslated region interacting with a cellular protein in human chondrosarcoma cells. AB - Connective tissue growth factor (CTGF/CCN2) can be induced by various forms of stress such as exposure to high glucose, mechanical load, or hypoxia. Here, we investigated the molecular mechanism involved in the induction of ctgf/ccn2 by hypoxia in a human chondrosarcoma cell line, HCS-2/8. Hypoxia increased the ctgf/ccn2 mRNA level by altering the 3'-untranslated region (UTR)-mediated mRNA stability without requiring de novo protein synthesis. After a series of extensive analyses, we eventually found that the cis-repressive element of 84 bases within the 3'-UTR specifically bound to a cytoplasmic/nuclear protein. By conducting a UV crosslinking assay, we found the cytoplasmic/nuclear protein to be a 35 kDa molecule that bound to the cis-element in a hypoxia-inducible manner. These results suggest that a cis-element in the 3'-UTR of ctgf/ccn2 mRNA and trans-factor counterpart(s) play an important role in the post-transcriptional regulation by determining the stability of ctgf/ccn2 mRNA. PMID- 16247470 TI - Hypoxia selects for high-metastatic Lewis lung carcinoma cells overexpressing Mcl 1 and exhibiting reduced apoptotic potential in solid tumors. AB - Low oxygen tension (hypoxia) is a common feature of solid tumors and stimulates the expressions of a variety of genes including those related to angiogenesis, apoptosis and endoplasmic reticulum (ER) stress response. Here we show a close correlation between metastatic potential and the resistance to hypoxia- and ER stress-induced apoptosis among the cell lines with differing metastatic potential derived from Lewis lung carcinoma. An apoptosis-specific expression profiling and immunoblot analyses revealed that the expression of antiapoptotic Mcl-1 increased as the resistance to apoptosis increased. Downregulation of the Mcl-1 expression in the high-metastatic cells by Mcl-1 small interfering RNA increased the sensitivity to hypoxia-induced apoptosis and decreased the metastatic ability. The hypoxia-induced apoptosis was not associated with p53 accumulation, although at present it is not possible to conclude that apoptosis-induced apoptosis is p53 independent. There was no correlation between the expression levels of ER stress response proteins GADD153, GRP78 and ORP150 and the resistance to hypoxia or ER stresses. In vitro, small numbers of the high-metastatic cells overtook the low metastatic cells after exposure to several rounds of hypoxia and reoxygenation. In solid tumors initially established from equal mixtures, the proportion of the high-metastatic cells to low-metastatic cells was significantly higher in hypoxic areas. Moreover, the high-metastatic cells were overtaking the low-metastatic cells in some of the tumors. Thus, tumor hypoxia and ER stress may provide a physiological selective pressure for the expansion of the high-metastatic cells overexpressing Mcl-1 and exhibiting reduced apoptotic potential in solid tumors. PMID- 16247468 TI - Hair cycle and wound healing in mice with a keratinocyte-restricted deletion of FAK. AB - Focal adhesion kinase (FAK) is a critical component in transducing signals downstream of both integrins and growth factor receptors. To determine how the loss of FAK affects the epidermis in vivo, we have generated a mouse model with a keratinocyte-restricted deletion of fak (FAKK5 KO mice). FAK(K5 KO) mice displayed three major phenotypes--irregularities of hair cycle, sebaceous glands hypoplasia, and a thinner epidermis--pointing to defects in the proliferative capacity of multipotent stem cells found in the bulge. FAK-null keratinocytes in conventional primary culture undergo massive apoptosis hindering further analyses, whereas the defects observed in vivo do not shorten the mouse lifespan. These results suggest that the structure and the signaling environment of the native tissue may overcome the lack of signaling through FAK. Our findings point to the importance of in vivo and three-dimensional in vitro models in analyses of cell migration, proliferation, and survival. Surprisingly, the difference between FAKloxP/+ and FAKK5 KO mice in wound closure was not statistically significant, suggesting that in vivo loss of FAK does not affect migration/proliferation of basal keratinocytes in the same way as it affects multipotent stem cells of the skin. PMID- 16247472 TI - Phosphorylation of the cytokinesis regulator ECT2 at G2/M phase stimulates association of the mitotic kinase Plk1 and accumulation of GTP-bound RhoA. AB - The epithelial cell transforming gene 2 (ECT2) protooncogene encodes a Rho exchange factor, and regulates cytokinesis. ECT2 is phosphorylated in G2/M phases, but its role in the biological function is not known. Here we show that two mitotic kinases, Cdk1 and polo-like kinase 1 (Plk1), phosphorylate ECT2 in vitro. We identified an in vitro Cdk1 phosphorylation site (T412) in ECT2, which comprises a consensus phosphospecific-binding module for the Plk1 polo-box domain (PBD). Endogenous ECT2 in mitotic cells strongly associated with Plk1 PBD, and this binding was inhibited by phosphatase treatment. A phosphorylation-deficient mutant form of ECT2, T412A, did not exhibit strong association with Plk1 PBD compared with wild-type (WT) ECT2. Moreover, ECT2 T412A, but not phosphomimic T412D, displayed a diminished accumulation of GTP-bound RhoA compared with WT ECT2, suggesting that phosphorylation of Thr-412 is critical for the catalytic activity of ECT2. Moreover, while overexpression of WT ECT2 or the T412D mutant caused cortical hyperactivity in U2OS cells during cell division, this activity was not observed in cells expressing ECT2 T412A. These results suggest that ECT2 is regulated by Cdk1 and Plk1 in concert. PMID- 16247471 TI - Dissection of transcriptional and non-transcriptional p53 activities in the response to genotoxic stress. AB - Following genotoxic stress, p53 either rescues a damaged cell or promotes its elimination. The parameters determining a specific outcome of the p53 response are largely unknown. In mouse fibroblasts treated with different irradiation schemes, we monitored transcriptional and non-transcriptional p53 activities and identified determinants that initiate an anti- or a pro-apoptotic p53 response within the context of p53-independent stress signaling. The primary, transcription-mediated p53 response in these cells is anti-apoptotic, while induction of p53-dependent apoptosis requires an additional, transcription independent p53 activity, provided by high intracellular levels of activated p53. High intracellular levels of p53 were selectively generated after apoptosis inducing high-dose UV-irradiation, and correlated with a strongly delayed upregulation of Mdm2. Following high-dose UV-irradiation, p53 accumulated in the cytoplasm and led to activation of the pro-apoptotic protein Bax. As p53 dependent Bax-activation is transcription-independent, we postulated that certain transcription-deficient mutant p53 proteins might also exert this activity. Indeed we found an endogenous, transcription-inactive mutant p53 that upon genotoxic stress induced Bax-activation in vivo. Our results demonstrate the impact and in vivo relevance of non-transcriptional mechanisms for wild-type and mutant p53-mediated apoptosis. PMID- 16247473 TI - RING finger-dependent ubiquitination by PRAJA is dependent on TGF-beta and potentially defines the functional status of the tumor suppressor ELF. AB - In gastrointestinal cells, biological signals for transforming growth factor-beta (TGF-beta) are transduced through transmembrane serine/threonine kinase receptors that signal to Smad proteins. Smad4, a tumor suppressor, is often mutated in human gastrointestinal cancers. The mechanism of Smad4 inactivation, however, remains uncertain and could be through E3-mediated ubiquitination of Smad4/adaptor protein complexes. Disruption of ELF (embryonic liver fodrin), a Smad4 adaptor protein, modulates TGF-beta signaling. We have found that PRAJA, a RING-H2 protein, interacts with ELF in a TGF-beta-dependent manner, with a fivefold increase of PRAJA expression and a subsequent decrease in ELF and Smad4 expression, in gastrointestinal cancer cell lines (P < 0.05). Strikingly, PRAJA manifests substantial E3-dependent ubiquitination of ELF and Smad3, but not Smad4. Delta-PRAJA, which has a deleted RING finger domain at the C terminus, abolishes ubiquitination of ELF. A stable cell line that overexpresses PRAJA exhibits low levels of ELF in comparison to a Delta-PRAJA stable cell line, where ELF expression is high compared to normal controls. The alteration of ELF and/or Smad4 expression and/or function in the TGF-beta signaling pathway may be induced by enhancement of ELF degradation, which is mediated by a high-level expression of PRAJA in gastrointestinal cancers. In hepatocytes, half-life (t(1/2)) and rate constant for degradation (k(D)) of ELF is 1.91 h and 21.72 min(-1) when coupled with ectopic expression of PRAJA in cells stimulated by TGF-beta, compared to PRAJA-transfected unstimulated cells (t(1/2) = 4.33 h and k(D) = 9.6 min(-1)). These studies reveal a mechanism for tumorigenesis whereby defects in adaptor proteins for Smads, such as ELF, can undergo degradation by PRAJA, through the ubiquitin-mediated pathway. PMID- 16247474 TI - c-FLIP inhibits chemotherapy-induced colorectal cancer cell death. AB - c-FLIP inhibits caspase 8 activation and apoptosis mediated by death receptors such as Fas and DR5. We studied the effect of c-FLIP on the apoptotic response to chemotherapies used in colorectal cancer (CRC) (5-fluorouracil, oxaliplatin and irinotecan). Simultaneous downregulation of both c-FLIP splice forms c-FLIP(L) and c-FLIP(S) with siRNA synergistically enhanced chemotherapy-induced apoptosis in p53 wild-type (HCT116p53(+/+), RKO), null (HCT116p53(-/-)) and mutant (H630) CRC cell lines. Furthermore, overexpression of c-FLIP(L), but not c-FLIP(S), potently inhibited apoptosis induced by chemotherapy in HCT116p53(+/+) cells, suggesting that c-FLIP(L) was the more important splice form in mediating chemoresistance. In support of this, siRNA specifically targeted against c FLIP(L) synergistically enhanced chemotherapy-induced apoptosis in a manner similar to the siRNA targeted against both splice forms. Inhibition of caspase 8 blocked the enhanced apoptosis induced by c-FLIP-targeted (FT) siRNA and chemotherapy. Furthermore, we found that downregulating cell surface DR5, but not Fas, also inhibited apoptosis induced by FT siRNA and chemotherapy. Interestingly, these effects were not dependent on activation of DR5 by its ligand TRAIL. These results indicate that c-FLIP inhibits TRAIL-independent, DR5- and caspase 8-dependent apoptosis in response to chemotherapy in CRC cells. Moreover, targeting c-FLIP in combination with existing chemotherapies may have therapeutic potential for the treatment of CRC. PMID- 16247475 TI - The cytoskeletal network controls c-Jun translation in a UTR-dependent manner. AB - The cytoskeleton is a dynamic network that undergoes restructuring during various cellular events, influencing cell proliferation, differentiation, and apoptosis. Here, we report that accumulation of c-Jun, a member of the AP1 family of transcription factors that play a key role in normal and aberrant cell growth, dramatically increases upon depolymerization of the cytoskeleton, and that, unexpectedly, this increase is controlled translationally. Depolymerization of the actin or microtubule network induces an increase in c-Jun accumulation with no corresponding increase in c-Jun mRNA or in the half-life of the c-Jun protein, but rather in the translatability of its transcript. This increase is mediated by the untranslated regions (UTRs) of c-Jun mRNA, and is not dependent on activated mitogen-activated protein kinase pathways. This novel mechanism of c-Jun regulation might be relevant to physiological conditions in which c-Jun plays a pivotal role. PMID- 16247477 TI - The hepatitis B virus X protein enhances AP-1 activation through interaction with Jab1. AB - Hepatitis B virus X protein (HBx) has many cellular functions and is a major factor in hepatitis and hepatocellular carcinoma caused by HBV infection. A proteomic approach was used to search for HBx-interacting proteins in order to elucidate the molecular mechanism of hepatocarcinogenesis. HBx was attached to myc and flag tags (MEF tags) and expressed in 293T cells; the protein complex formed within the cells was purified and characterized by mass spectrometry. COP9 signalosome (CSN) subunits 3 and 4 were subsequently identified as HBx interacting proteins. In addition, CSN subunit 5, Jun activation domain-binding protein 1 (Jab1), was shown to be a novel cellular target of HBx. In vivo and in vitro interactions between HBx and Jab1 were confirmed by standard immunoprecipitation and GST pull-down assays. An analysis of HBx deletion constructs showed that amino acids 30-125 of HBx were responsible for binding to Jab1. Confocal laser microscopy demonstrated that HBx was mainly localized in the cytoplasm, while Jab1 was found mainly in the nucleus and partially in the cytoplasm, and that the two proteins colocalized in the cytoplasm. The cotransfection of HBx and Jab1 resulted in substantial activator protein 1 (AP-1) activation and knockdown of endogenous Jab1 attenuated AP-1 activation caused by HBx. In addition, the coexpression of HBx and Jab1 potentiated phosphorylation of JNK, leading to the subsequent phosphorylation of c-Jun, whereas the level of c Jun and JNK phosphorylation induced by HBx was decreased in Jab1 knockdown cells. These results suggest that the interaction between HBx and Jab1 enhances HBx mediated AP-1 activation. PMID- 16247476 TI - Bone morphogenetic protein 2 stimulation of tumor growth involves the activation of Smad-1/5. AB - Morphogenetic protein 2 (BMP-2) is normally expressed in the embryo promoting the development of several organs. Aberrant expression of BMP-2 occurs in approximately 98% of lung carcinomas, however, its role in regulating tumor growth is poorly understood. We show that BMP-2 induces Id-1 expression in lung cancer cell lines through its activation of Smad-1/5, which is dependent on cell culture conditions. A549 cells in DMEM/5% FCS BMP-2 activated Smad-1/5 and caused a transient increase in proliferation. In serum-free medium, BMP-2 induced significantly less Smad-1/5 activation and Id-1 expression, and produced significant growth inhibition. The affect of BMP-2 on tumor growth in vivo was substantially more significant. Recombinant BMP-2 coinjected with A549 cells, into nude mice increased proliferation and produced an increase in Id-1 expression. Forced expression of BMP-2 in A549 cells significantly enhanced tumor growth in the lungs following intravenous injection but not of subcutaneous tumors. Tumors in the lung were found to have an activated Smad-1/5 and expressed Id-1. Subcutaneous tumors expressed less activated Smad-1/5 and Id-1 than that of controls. Human lung carcinomas were also found to express an activated Smad-1/5 and Id-1. We provide evidence that BMP-2 promotes tumor growth. This paper highlights that cell culture experiments may not reveal the full biological affects of BMP-2, and its activity varies depending of the local environment. PMID- 16247478 TI - Transcriptional responses to ionizing radiation reveal that p53R2 protects against radiation-induced mutagenesis in human lymphoblastoid cells. AB - The p53 protein has been implicated in multiple cellular responses related to DNA damage. Alterations in any of these cellular responses could be related to increased genomic instability. Our previous study has shown that mutations in p53 lead to hypermutability to ionizing radiation. To investigate further how p53 is involved in regulating mutational processes, we used 8K cDNA microarrays to compare the patterns of gene expression among three closely related human cell lines with different p53 status including TK6 (wild-type p53), NH32 (p53-null), and WTK1 (mutant p53). Total RNA samples were collected at 1, 3, 6, 9, and 24 h after 10 Gy gamma-irradiation. Template-based clustering analysis of the gene expression over the time course showed that 464 genes are either up or downregulated by at least twofold following radiation treatment. In addition, cluster analyses of gene expression profiles among these three cell lines revealed distinct patterns. In TK6, 165 genes were upregulated, while 36 genes were downregulated. In contrast, in WTK1 75 genes were upregulated and 12 genes were downregulated. In NH32, only 54 genes were upregulated. Furthermore, we found several genes associated with DNA repair namely p53R2, DDB2, XPC, PCNA, BTG2, and MSH2 that were highly induced in TK6 compared to WTK1 and NH32. p53R2, which is regulated by the tumor suppressor p53, is a small subunit of ribonucleotide reductase. To determine whether it is involved in radiation induced mutagenesis, p53R2 protein was inhibited by siRNA in TK6 cells and followed by 2 Gy radiation. The background mutation frequencies at the TK locus of siRNA-transfected TK6 cells were about three times higher than those seen in TK6 cells. The mutation frequencies of siRNA-transfected TK6 cells after 2 Gy radiation were significantly higher than the irradiated TK6 cells without p53R2 knock down. These results indicate that p53R2 was induced by p53 protein and is involved in protecting against radiation-induced mutagenesis. PMID- 16247479 TI - Overexpression of CEBPbeta correlates with decreased TFF1 in gastric cancer. AB - CCAAT element binding protein beta (C/EBPbeta) is an important regulator of cell growth, differentiation and in promoting tumor invasiveness. C/EBPbeta is located on chromosome 20q, which is amplified in many solid tumors including gastric cancers (GC). We sought to characterize the status of C/EBPbeta expression in GCs, which was recently found to repres TFF1 gene. Microarray analysis revealed overexpression of C/EBPbeta in 25 of 27 (93%) GC when compared to 12 normal gastric tissue samples. RT-PCR analysis confirmed the overexpression of C/EBPbeta transcripts in 54 of 59 (91%) GC. In total, 15 of 18 gastric tumors exhibited at least fivefold higher C/EBPbeta transcript levels compared to their corresponding adjacent normal gastric tissue samples. Moreover, immunohistochemistry analysis demonstrated increased nuclear staining of C/EBPbeta in 10 of 13 GC and at least fourfold overexpression of C/EBPbeta in three primary GC compared to adjacent normal gastric tissue. Furthermore, a striking correlation of decreased TFF1 expression with increased C/EBPbeta was observed in the gastric tumors studied. Microarray analysis demonstrated a loss of TFF1 expression in all 27 GC cases examined, of which 25 exhibited high C/EBPbeta expression compared to normal gastric tissue. RT-PCR analysis revealed loss of TFF1 expression in 56 of 59 gastric tumors in which 54 of these tumors exhibited overexpression of C/EBPbeta. Immunohistochemical analysis revealed overexpression of C/EBPbeta in 10 of 13 gastric tumors that exhibited low expression of TFF1 at the protein level. Thus, overexpression of the transcription factor C/EBPbeta in the majority of GCs is a novel finding. PMID- 16247480 TI - Type I collagen is a molecular target for inhibition of angiogenesis by endogenous thrombospondin-1. AB - Three-dimensional explant cultures of muscle tissue were used to characterize secreted proteins regulated by endogenous levels of the angiogenesis modulator thrombospondin (TSP)-1. Explants from TSP1 null mice exhibit enhanced neovascularization associated with increased endothelial outgrowth but decreased outgrowth of perivascular smooth muscle cells . The absence of endogenous TSP1 did not diminish activation of latent transforming growth factor-beta and moderately decreased matrix metalloproteinase levels. However, significant changes in other secreted proteins were observed. Endogenous TSP1 decreased mRNA levels for collagens Ialpha1, Ialpha2, and IIIalpha1 and laminin alpha4 and increased collagen IValpha1 mRNA expression. Endogenous TSP1 also decreased the level of type I collagen protein produced by the vascular outgrowths. Collagens Ialpha1, Ialpha2, and IIIalpha1 are known tumor endothelial markers, suggesting that TSP1 coordinately regulates a set of extracellular matrix genes that reverse the angiogenic switch. Suppression of collagen Ialpha1 or Ialpha2 mRNAs using antisense morpholinos inhibited outgrowth in TSP1 null explants and proliferation of TSP1 null endothelial cells, indicating that type I collagen synthesis is limiting for this neovascularization response. PMID- 16247481 TI - SWI/SNF complex is essential for NRSF-mediated suppression of neuronal genes in human nonsmall cell lung carcinoma cell lines. AB - Mammalian chromatin remodeling factor, SWI/SNF complex contains a single molecule of either Brm or BRG1 as the ATPase catalytic subunit. Here, we show that the SWI/SNF complex forms a larger complex with neuron-restrictive silencer factor (NRSF) and its corepressors, mSin3A and CoREST, in human nonsmall cell lung carcinoma cell lines. We also demonstrate that the strong transcriptional suppression of such neuron-specific genes as synaptophysin and SCG10 by NRSF in these non-neural cells requires the functional SWI/SNF complex; these neuronal genes were elevated in cell lines deficient in both Brm and BRG1, whereas retrovirus vectors expressing siRNAs targeting integral components of SWI/SNF complex (Brm/BRG1 or Ini1) induced expression of these neuronal genes in SWI/SNF competent cell lines. In cell lines deficient in both Brm and BRG1, exogenous Brm or BRG1 suppressed expression of these neuronal genes in an ATP-dependent manner and induced efficient and specific deacetylation of histone H4 around the NRSF binding site present in the synaptophysin gene by a large complex containing the recruited functional SWI/SNF complex. Patients with Brm/BRG1-deficient lung carcinoma have been reported to carry poor prognosis; derepression of NRSF regulated genes including these neuron-specific genes could contribute to enhance tumorigenicity and also would provide selective markers for Brm/BRG1-deficient tumors. PMID- 16247483 TI - Identification of novel tumour-associated genes differentially expressed in the process of squamous cell cancer development. AB - Chemically induced mouse skin carcinogenesis represents the most extensively utilized animal model to unravel the multistage nature of tumour development and to design novel therapeutic concepts of human epithelial neoplasia. We combined this tumour model with comprehensive gene expression analysis and could identify a large set of novel tumour-associated genes that have not been associated with epithelial skin cancer development yet. Expression data of selected genes were confirmed by semiquantitative and quantitative RT-PCR as well as in situ hybridization and immunofluorescence analysis on mouse tumour sections. Enhanced expression of genes identified in our screen was also demonstrated in mouse keratinocyte cell lines that form tumours in vivo. Self-organizing map clustering was performed to identify different kinetics of gene expression and coregulation during skin cancer progression. Detailed analysis of differential expressed genes according to their functional annotation confirmed the involvement of several biological processes, such as regulation of cell cycle, apoptosis, extracellular proteolysis and cell adhesion, during skin malignancy. Finally, we detected high transcript levels of ANXA1, LCN2 and S100A8 as well as reduced levels for NDR2 protein in human skin tumour specimens demonstrating that tumour-associated genes identified in the chemically induced tumour model might be of great relevance for the understanding of human epithelial malignancies as well. PMID- 16247482 TI - LMP1 signaling and activation of NF-kappaB in LMP1 transgenic mice. AB - Transgenic mice expressing Epstein-Barr virus (EBV) latent membrane protein 1 (LMP1) under the control of an immunoglobulin heavy-chain promoter and enhancer develop lymphoma at a threefold higher incidence than LMP1-negative mice. In vitro, LMP1 activates numerous signaling pathways including p38, c-Jun N terminal kinase (JNK), phosphatidylinositol 3 kinase (PI3K)/Akt, and NF-kappaB through interactions with tumor necrosis receptor-associated factors (TRAFs). These pathways are frequently activated in EBV-associated malignancies, although their activation cannot be definitively linked to LMP1 expression in vivo. In this study, interactions between LMP1 and TRAFs and the activation of PI3K/Akt, JNK, p38, and NF-kappaB were examined in LMP1 transgenic mice. LMP1 co immunoprecipitated with TRAFs 1, 2, and 3. Akt, JNK, and p38 were activated in LMP1-positive and -negative splenocytes as well as LMP1-positive and -negative lymphomas. Multiple forms of NF-kappaB were activated in healthy splenocytes from LMP1 transgenic mice, in contrast to healthy splenocytes from LMP1-negative mice. However, in both LMP1-positive and -negative lymphomas, only the oncogenic NF kappaB c-Rel, was specifically activated. Similarly to EBV-associated malignancies, p53 protein was detected at high levels in the transgenic lymphomas, although mutations were not detected in the p53 gene. These data indicate that NF-kappaB is activated in LMP1-positive healthy splenocytes; however, NF-kappaB c-Rel is specifically activated in both the transgenic lymphomas and in the rare lymphomas that develop in negative mice. The LMP1 mediated activation of NF-kappaB may contribute to the specific activation of c Rel and lead to the increased development of lymphoma in the LMP1 transgenic mice. PMID- 16247484 TI - Epigenetic silencing of AXIN2 in colorectal carcinoma with microsatellite instability. AB - Mutation or epigenetic silencing of mismatch repair genes, such as MLH1 and MSH2, results in microsatellite instability (MSI) in the genome of a subset of colorectal carcinomas (CRCs). However, little is yet known of genes that directly contribute to tumor formation in such cancers. To characterize MSI-dependent changes in gene expression, we have now compared transcriptomes between fresh CRC specimens positive or negative for MSI (n=10 for each) with the use of high density oligonucleotide microarrays harboring >44,000 probe sets. Correspondence analysis of the expression patterns of isolated MSI-associated genes revealed that the transcriptome of MSI+ CRCs is clearly distinct from that of MSI- CRCs. Such MSI-associated genes included that for AXIN2, an important component of the WNT signaling pathway. AXIN2 was silenced, apparently as a result of extensive methylation of its promoter region, specifically in MSI+ CRC specimens. Forced expression of AXIN2, either by treatment with 5'-azacytidine or by transfection with AXIN2 cDNA, resulted in rapid cell death in an MSI+ CRC cell line. These data indicate that epigenetic silencing of AXIN2 is specifically associated with carcinogenesis in MSI+ CRCs. PMID- 16247485 TI - Differential regulation of different human papilloma virus variants by the POU family transcription factor Brn-3a. AB - The Brn-3a POU family transcription factor is overexpressed in human cervical carcinoma biopsies and is able to activate expression of the human papilloma virus type 16 (HPV-16) upstream regulatory region (URR), which drives the expression of the E6 and E7 oncoproteins. Inhibition of Brn-3a expression in human cervical cancer cells inhibits HPV gene expression and reduces cellular growth and anchorage independence in vitro as well as the ability to form tumours in vivo. Here, we show that Brn-3a differentially regulates different HPV-16 variants that have previously been shown to be associated with different risks of progression to cervical carcinoma. In human cervical material, Brn-3a levels correlate directly with HPV E6 levels in individuals infected with a high risk variant of HPV-16, whereas this is not the case for a low-risk variant. Moreover, the URRs of high- and intermediate-risk variants are activated by Brn-3a in transfection assays, whereas the URR of a low-risk variant is not. The change of one or two bases in a low-risk variant URR to their equivalent in a higher-risk URR can render the URR responsive to Brn-3a and vice versa. These results help explain why the specific interplay between viral and cellular factors necessary for the progression to cervical carcinoma only occurs in a minority of those infected with HPV-16. PMID- 16247486 TI - LIGHT induces differentiation of mouse embryonic stem cells associated with activation of ERK5. AB - LT-related inducible ligand that competes for glycoprotein D binding to herpesvirus entry mediator on T cells (LIGHT) is a recently cloned type II transmembrane protein belonging to the TNF family that was originally identified as a weak inducer of apoptosis. This cytokine has been extensively defined in its role on T-cell regulation and dendritic cell maturation. However, whether this cytokine regulates stem cell proliferation and/or differentiation remains unknown. In this study, we transduced exogenous LIGHT into embryonic stem cells (ES cells) and found it induced their differentiation. The expression of phospho STAT3, Nanog and Oct-4 was reduced in LIGHT-transduced ES cells compared with wild-type ES cells. LIGHT-transduced ES cells exhibit a low level of SSEA-1 surface antigen and alkaline phosphatase staining compared with wild-type cells. Introduction of LIGHT into ES cells results in the dephosphorylation of MKP-3 and activation of extracellular signal-regulated kinase (ERK)5. When ERK5 was inhibited by the specific inhibitor PD184352 or knocked down by ERK5 siRNA, reduction of Oct-4 and SSEA-1 expression was rescued. We conclude that LIGHT overrides Leukemia inhibitory factor to induce ES cell differentiation associated with activation of ERK5. PMID- 16247487 TI - Increased susceptibility to apoptosis in CD45(+) myeloma cells accompanied by the increased expression of VDAC1. AB - Expression of CD45 is quite variable in human myeloma cells and cell lines, such as U266, and CD45(+) U266 proliferates in response to a growth factor, interleukin-6. Here, we show that CD45(+) myeloma cell lines were more sensitive to various apoptotic stimuli, such as oxidative stress and endoplasmic reticulum (ER)-stress, than CD45(-) cells. Reactive oxygen species and calcium ion seemed to be involved in the susceptibility to apoptosis of CD45(+) U266. The activation of the src family kinases associated with CD45 phosphatase played an important role in the augmented apoptosis in CD45(+) U266 by oxidative stress. These results indicate that the CD45-expression renders myeloma cells competent for not only mitogenic but also apoptotic stimuli, resulting in either proliferation or apoptosis of CD45(+) myeloma cells dependently upon the circumstantial stimuli. Furthermore, voltage-dependent anion channel (VDAC) 1 was identified as a gene highly expressed in CD45(+) U266 by cDNA subtraction. The increased expression of VDAC1 seemed to augment the sensitivity to the ER-stress because the VDAC1 transfected U266 was more susceptible to the thapsigargin-induced apoptosis. Thus, CD45 expression accompanied by the increased VDAC1 expression sensitizes myeloma cells to the various extracellular stimuli that trigger apoptosis via the mitochondrial pathways. PMID- 16247488 TI - A novel protein isoform of catechol O-methyltransferase (COMT): brain expression analysis in schizophrenia and bipolar disorder and effect of Val158Met genotype. PMID- 16247489 TI - Polymorphisms in the prostate-specific antigen gene promoter do not predict serum prostate-specific antigen levels in African-American men. AB - A major problem with the use of serum prostate-specific antigen (PSA) in predicting prostate cancer risk is the considerable variability of such measurements. Cramer et al. identified a set of single-nucleotide polymorphisms (SNPs) in the upstream regulatory region of the PSA gene that were each associated with increased promoter activity and serum PSA, further suggesting that genotyping these SNPs could be useful in improving the predictive value of PSA screening. In order to replicate this finding, DNA samples from 475 African American men were genotyped for the same SNPs and no association was observed with either serum PSA level or prostate cancer diagnosis. PMID- 16247490 TI - How many ways to die? How many different models of cell death? PMID- 16247491 TI - Classification of cell death: recommendations of the Nomenclature Committee on Cell Death. PMID- 16247492 TI - Doctor Jekyll and Mister Hyde: autophagy can promote both cell survival and cell death. PMID- 16247493 TI - Anoikis. PMID- 16247494 TI - Mitochondrial permeability transition in apoptosis and necrosis. PMID- 16247495 TI - Pathogens and autophagy: subverting to survive. PMID- 16247496 TI - Macroautophagy versus mitochondrial autophagy: a question of fate? PMID- 16247498 TI - Autophagy and signaling: their role in cell survival and cell death. AB - Macroautophagy is a vacuolar, self-digesting mechanism responsible for the removal of long-lived proteins and damaged organelles by the lysosome. The discovery of the ATG genes has provided key information about the formation of the autophagosome, and about the role of macroautophagy in allowing cells to survive during nutrient depletion and/or in the absence of growth factors. Two connected signaling pathways encompassing class-I phosphatidylinositol 3-kinase and (mammalian) target of rapamycin play a central role in controlling macroautophagy in response to starvation. However, a considerable body of literature reports that macroautophagy is also a cell death mechanism that can occur either in the absence of detectable signs of apoptosis (via autophagic cell death) or concomitantly with apoptosis. Macroautophagy is activated by signaling pathways that also control apoptosis. The aim of this review is to discuss the signaling pathways that control macroautophagy during cell survival and cell death. PMID- 16247497 TI - Death penalty for keratinocytes: apoptosis versus cornification. AB - Homeostasis implies a balance between cell growth and cell death. This balance is essential for the development and maintenance of multicellular organisms. Homeostasis is controlled by several mechanisms including apoptosis, a process by which cells condemned to death are completely eliminated. However, in some cases, total destruction and removal of dead cells is not desirable, as when they fulfil a specific function such as formation of the skin barrier provided by corneocytes, also known as terminally differentiated keratinocytes. In this case, programmed cell death results in accumulation of functional cell corpses. Previously, this process has been associated with apoptotic cell death. In this overview, we discuss differences and similarities in the molecular regulation of epidermal programmed cell death and apoptosis. We conclude that despite earlier confusion, apoptosis and cornification occur through distinct molecular pathways, and that possibly antiapoptotic mechanisms are implicated in the terminal differentiation of keratinocytes. PMID- 16247499 TI - Immune surveillance of intracellular pathogens via autophagy. AB - MHC class II molecules are thought to present peptides derived from extracellular proteins to CD4+ T cells, which are important mediators of adaptive immunity to infections. In contrast, autophagy delivers constitutively cytosolic material for lysosomal degradation and has so far been recognized as an efficient mechanism of innate immunity against bacteria and viruses. Recent studies, however, link these two pathways and suggest that intracellular cytosolic and nuclear antigens are processed for MHC class II presentation after autophagy. PMID- 16247500 TI - Another way to die: autophagic programmed cell death. AB - Programmed cell death (PCD) is one of the important terminal paths for the cells of metazoans, and is involved in a variety of biological events that include morphogenesis, maintenance of tissue homeostasis, and elimination of harmful cells. Dysfunction of PCD leads to various diseases in humans, including cancer and several degenerative diseases. Apoptosis is not the only form of PCD. Recent studies have provided evidence that there is another mechanism of PCD, which is associated with the appearance of autophagosomes and depends on autophagy proteins. This form of cell death most likely corresponds to a process that has been morphologically defined as autophagic PCD. The present review summarizes recent experimental evidence about autophagic PCD and discusses some aspects of this form of cell death, including the mechanisms that may distinguish autophagic death from the process of autophagy involved in cell survival. PMID- 16247501 TI - The pleiotropic role of autophagy: from protein metabolism to bactericide. AB - Autophagy is in principle a nonselective, bulk degradation system within cells, with a contribution to intracellular protein degradation estimated to be as large as that of the ubiquitin--proteasome system. The primary roles of autophagy are baseline turnover of intracellular proteins and organelles, production of amino acids in nutrient emergency, and regression of retired tissues. These functions guarantee rejuvenation and adaptation to adverse conditions, and even underlie dynamic processes such as development/metamorphosis. In addition, several other roles for autophagy have recently been discovered, such as presentation of endogenous antigens and degradation of invasive bacteria. This review will discuss the biological significance of autophagy from yeast to higher eukaryotes. PMID- 16247504 TI - Subcutaneous administration of ghrelin stimulates energy intake in healthy lean human volunteers. AB - BACKGROUND: The gastric hormone ghrelin appears a useful agent to stimulate food intake in people with anorexia of illness. The loss of ghrelin's acyl group renders it inactive, thus it has been thought that subcutaneous administration may be problematic. OBJECTIVE: To investigate whether human subjects are sensitive to the effects of ghrelin administered by single subcutaneous injection. STUDY DESIGN: Randomized, double-blind, placebo-controlled trial. SUBJECTS: Sixteen healthy lean volunteers (eight men and eight women). PROTOCOL: Fasted subjects received subcutaneous injections of ghrelin (3.6 nmol/kg) or saline. After 30 min, a buffet breakfast was served. RESULTS: Ghrelin injection increased energy intake by 27% (ghrelin 5076 +/- 691 kJ versus saline 4230+/-607 kJ, P = 0.04). Ghrelin appeared to enhance the perceived palatability of the food offered (palatability score: ghrelin 81.1 +/- 3.6 versus saline 70.0 +/- 4.4; P = 0.03). CONCLUSIONS: These results suggest that subcutaneous ghrelin is effective at stimulating energy intake and improving palatability and may be of direct use in the treatment of appetite loss. PMID- 16247505 TI - Relationship of obesity to job stress and eating behavior in male Japanese workers. AB - OBJECTIVE: To examine a possible relationship between obesity, job stress, and eating behavior in male Japanese workers. METHOD: A questionnaire on life style, job stress, and eating behavior was conducted with 208 male workers aged 19-60 years (33.7+/-12.3 years) in a manufacturing industry in Japan. Height and weight were measured in an annual health examination. The relation between obesity, job stress, and eating behavior were analyzed between 141 nonobese subjects (BMI or=25.0 kg/m(2)). RESULTS: Obesity was associated with psychological stress responses of tension/anxiety, especially tension. Tension/anxiety was also related to job demands positively and job latitudes negatively. The eating behaviors of subjects with tension/anxiety resembled those of the obese subjects. CONCLUSIONS: The present study suggests that obese male Japanese workers tend to be in a stressful state from high job demands and low job latitudes in the workplace. Such stressful conditions may affect eating behaviors to eat much and contribute to obesity. Stress management might be necessary in the workplace for the prevention of obesity among male Japanese workers. PMID- 16247502 TI - Autophagy: molecular machinery for self-eating. AB - Autophagy is a highly conserved process in eukaryotes in which the cytoplasm, including excess or aberrant organelles, is sequestered into double-membrane vesicles and delivered to the degradative organelle, the lysosome/vacuole, for breakdown and eventual recycling of the resulting macromolecules. This process has an important role in various biological events such as adaptation to changing environmental conditions, cellular remodeling during development and differentiation, and determination of lifespan. Auto-phagy is also involved in preventing certain types of disease, although it may contribute to some pathologies. Recent studies have identified many components that are required to drive this complicated cellular process. Auto-phagy-related genes were first identified in yeast, but homologs are found in all eukaryotes. Analyses in a range of model systems have provided huge advances toward understanding the molecular basis of autophagy. Here we review our current knowledge on the machinery and molecular mechanism of autophagy. PMID- 16247507 TI - PPARgamma activity in subcutaneous abdominal fat tissue and fat mass gain during short-term overfeeding. AB - OBJECTIVE: As the peroxisome proliferator-activated receptor gamma (PPARgamma) plays a central role in fat mass regulation, we investigated whether initial subcutaneous PPARgamma activity is related to fat mass generation during overfeeding. SUBJECTS: Fourteen healthy female subjects (age 25 +/- 4 years, BMI 22.1 +/- 2.3 kg/m2). DESIGN AND MEASUREMENTS: Subjects were overfed with a diet supplying 50% more energy than baseline energy requirements for 14 days. Fasting blood samples were analyzed for leptin, insulin and glucose. Fasting subcutaneous abdominal fat biopsies were obtained for analysis of PPARgamma1, PPARgamma2, aP2 and UCP2 mRNAs. RESULTS: Initial PPARgamma1 and 2, aP2 and UCP2 mRNAs were not related to fat gain (P > 0.12). However, PPARgamma1, PPARgamma2 and aP2 mRNA changes were positively related to changes in plasma leptin (P < 0.05) and, except aP2 (P = 0.06), to fat gain (P < 0.05). PPARgamma and aP2 mRNA changes were positively related (P<0.01), indicating that PPARgamma mRNA levels reflected PPARgamma activity. CONCLUSION: These data suggest that the ability to increase PPARgamma activity might be involved in the susceptibility to gain weight during a positive energy balance. PMID- 16247506 TI - Promoter polymorphism in the macrophage migration inhibitory factor gene is associated with obesity. AB - OBJECTIVE: To explore the association of promoter polymorphisms of macrophage migration inhibitory factor (MIF) gene with obesity. SUBJECTS: In total, 213 nondiabetic Japanese subjects. They were divided into three groups according to World Health Organization definitions: lean (body mass index (BMI) <25 kg/m2), overweight (25 < or = BMI < 30 kg/m2) and obese (BMI> or = 30 kg/m2). METHODS: We examined two polymorphic loci in the MIF gene in the subjects: a single nucleotide polymorphism at position -173 (G/C) and a CATT-tetranucleotide repeat polymorphism at position -794, which both can affect promoter activity in different cells. RESULTS: We detected four alleles: 5-, 6-, 7- and 8-CATT at position -794. Genotypes without the 5-CATT allele (X/X, X refers to 6-, 7- or 8 CATT alleles) were more common in obese subjects than in lean or overweight groups (P = 0.013). The X-CATT allele was more frequent in obese subjects than in lean or overweight subjects (P = 0.030). In contrast, -173G/C was not associated with obesity. Among the haplotypes of the two promoter polymorphisms, G/5-CATT (( 173G/C)/(-794[CATT](5-8))) was associated with a decreased risk of obesity (P = 0.025) and G/6-CATT with an increased risk of overweight (P=0.028). CONCLUSION: Promoter polymorphism in the MIF gene is linked with obesity. PMID- 16247508 TI - Alpha2B-adrenergic receptor deletion polymorphism and cardiac autonomic nervous system responses to exercise in obese women. AB - OBJECTIVE: To investigate the association of short form (Glu9/Glu9) of the 12Glu9 deletion polymorphism of the alpha2B-adrenergic receptor (alpha2B-AR) gene polymorphism with the cardiac autonomic responsiveness during sustained isometric handgrip exercise. DESIGN: Cross-sectional clinical study. SUBJECTS: In all, 97 normotensive obese women (body mass index (BMI) = 33.2 kg/m2). Of these, 78 (80.41%) were genotyped as Glu12/Glu12, 13 (13.40%) as Glu12/Glu9 and six (6.19%) as Glu9/Glu9 form. MEASUREMENTS: The sympathovagal balance was assessed by means of power spectral analysis of heart rate variability at rest and during sustained isometric handgrip exercise at 30% of maximal voluntary handgrip contraction for 3 min. Two spectral components were analysed: low-frequency component reflecting sympathetic efferent activity and high-frequency power (HFnu) reflecting parasympathetic modulation. In addition, a normalized low-frequency power (LFnu) and HFnu were analysed. Genotypes were determined by polymerase chain reaction followed by agarose gel electrophoresis. RESULTS: There were no differences in baseline measurements among groups. The absolute level of LFnu throughout handgrip exercise was significantly lower in Glu9/Glu9 subjects compared with other genotypes, while the decline of absolute HFnu was significantly smaller compared with Glu12/Glu12 genotype. CONCLUSION: These findings suggest that 12Glu9 deletion polymorphism of the alpha2B-AR gene (Glu9/Glu9 genotype) might result in reduced autonomic responsiveness by altering cardiac sympathetic and vagal function during sustained handgrip exercise in normotensive obese women. PMID- 16247509 TI - Trends in resting pulse rate among students attending Glasgow University between 1948 and 1968: analyses of cross sectional surveys. AB - Evidence on long-term trends in physical activity is limited. We report that resting pulse rates--a proxy indicator of physical activity and fitness- increased among young adults attending Glasgow University between 1948 and 1968. PMID- 16247510 TI - Reduction of diet-induced obesity by a combination of tea-catechin intake and regular swimming. AB - OBJECTIVE: Obesity is a metabolic disorder resulting from imbalance between metabolizable energy intake and energy expenditure. It is known to be a strong risk factor for lifestyle-related diseases. Here, we investigated the effects of long-term intake of tea catechins (Cat) in combination with regular exercise (Ex) on the development of obesity in C57BL/6 mice. DESIGN: We compared body weight, adipose tissue mass, plasma parameters and beta-oxidation activity in mice fed a low-fat diet (5% triglyceride (TG); LF), a high-fat diet (30% TG; HF), a HF diet supplemented with 0.5% (w/w) tea Cat, a HF diet in addition to swimming Ex or a HF diet plus 0.5% tea Cat in addition to swimming Ex (Cat+Ex) for 15 weeks. Oxygen consumption and respiratory quotients were measured using indirect calorimetry. RESULTS: Tea-Cat intake in combination with swimming Ex suppressed HF diet-induced body-weight gain by 18 and 22%, respectively, compared to Ex and tea-Cat intake on their own. Visceral fat accumulation and the development of hyperinsulinemia and hyperleptinemia were also reduced in the HF+Cat+Ex group. Muscular beta-oxidation activity in this group was 69 and 52% higher, respectively, than that in the HF and HF+Cat groups. Lipid oxidation, determined using indirect calorimetry, was higher in the HF+Cat+Ex group, suggesting increased lipid utilization at the individual level. CONCLUSION: These results indicate that intake of tea Cat, together with regular Ex helps to reduce diet induced obesity. This effect might be attributed, at least in part, to the activation of whole-body energy metabolism. PMID- 16247511 TI - Lipids, lipoproteins, lifestyle, adiposity and fat-free mass during middle age: the Fels Longitudinal Study. AB - INTRODUCTION: Although lipid profiles tend to worsen with age, it is not fully known if such age-related changes are influenced primarily by body composition and lifestyle or by other aspects of aging. OBJECTIVE: We investigated the extent to which the fat and fat-free components of body mass index (BMI) and lifestyle factors influence patterns of change in lipids independent of age. DESIGN: Serial data were analyzed using sex-specific longitudinal models. These models use serial data from individuals to assume a general pattern of change over time, while allowing baseline age and the rate of change to vary among individuals. SUBJECTS: Serial data were obtained from 940 examinations of 269 healthy white participants (126 men, 143 women), aged 40-60 years, in the Fels Longitudinal Study. MEASUREMENTS: Measurements included age, the fat (FMI) and fat-free mass (FFMI) components of BMI, high-density lipoprotein (HDL-C), low-density lipoprotein (LDL-C), triglycerides (TG), total cholesterol (TC), fasting glucose and insulin, physical activity, alcohol use and smoking, and women's menopausal status and estrogen use. RESULTS: In both sexes, increased FMI was significantly associated with increased LDL-C, TG and TC, and decreased HDL-C. Increased FFMI was significantly related to decreased HDL-C and increased TG. Independent age effects remained significant only for LDL-C and TC in men and TC in women. Increased insulin was significantly related to increased TG in women. Moderate alcohol consumption was associated with higher HDL-C in men. Physical activity lowered male LDL-C and TC levels, and increased female HDL-C levels. Menopause was associated with increases in LDL-C. Premenopausal women not using estrogen had significantly lower HDL-C, TG, and TC than postmenopausal women taking estrogen. CONCLUSIONS: (1) Age is an important independent predictor for LDL-C and TC in men, and TC in women, but it is not as influential as body composition and lifestyle on HDL-C and TG in men and women, and LDL-C in women. (2) Increasing FMI is the major contributor to elevated TC, LDL-C and TG levels, and decreased HDL-C levels in men and women. (3) FFMI significantly influences HDL and TG levels in both sexes. (4) Maintaining a lower BMI via a reduced fat component may be more beneficial in lowering CVD risks than other factors. PMID- 16247513 TI - The role of metabotropic glutamate receptor 5 in learning and memory processes. AB - Metabotropic glutamate receptor 5 (mGluR5), a subtype in the group I mGluRs, couples to phospholipase C through Gq protein. Stimulation of mGluR5 leads to the release of calcium from intracellular stores and protein kinase C activation. In addition, links to different ion channels and other signaling mechanisms have also been revealed. MGluR5s are mainly localized postsynaptically on the periphery of synap-ses. MGluR5s have been implicated in synaptic plasticity and learning and memory. The development of the highly potent and selective mGluR5 antagonist 2-methyl-6-(phenylethynyl)-pyridine (MPEP) has facilitated the understanding of the roles of mGluR5s in the central nervous system. Both in vitro and in vivo studies have demonstrated that the activation of mGluR5s is necessary for some forms of long-term potentiation and long-term depression in different brain regions. Investigations of the effects of MPEP in various behavioral paradigms have concluded that mGluR5s play a critical role in aversive learning tasks and in hippocampal-dependent spatial learning. However, MPEP has proved ineffective in certain other learning tasks. MGluR5 knockout mice have shown impairments in water maze and radial arm maze performance as well as in contextual fear conditioning, but not in cue conditioning. This review summarizes recent advances reported on mGluR5 function in synaptic plasticity, learning and memory. The current development of positive and negative allosteric modulators of mGluR5 will provide new pharmacological tools to enhance our knowledge of these receptors in physiological and pathophysiological processes and will further facilitate new investigations on mGluR5 as a therapeutic target for a range of neurological and psychological disorders. PMID- 16247514 TI - Intraoral delivery of antimicrobials. AB - Antimicrobial therapies for oral diseases have been in use for centuries, but have undergone rapid changes in the last decade. In the coming years, antimicrobial strategies will become more sophisticated and efficacy will be greatly improved as new therapeutic technologies emerge. New delivery systems for common antimicrobials and novel agents to modulate the immune system, as well as biofilm formation and maturation, may be on the horizon. This review describes the development and the application of intraoral antimicrobial drug delivery in the oral environment. Current clinical uses of antimicrobials as well as future approaches to the treatment and prevention of oral infectious diseases are discussed. PMID- 16247515 TI - Targeting fatty acid synthase: potential for therapeutic intervention in her 2/neu-overexpressing breast cancer. AB - Fatty acid synthase (FAS)-catalyzed de novo fatty acid biosynthesis, an anabolic energy-storage pathway largely considered of minor importance in humans, actively contributes to the cancer phenotype by virtue of its ability to specifically regulate the expression and activity of Her-2/neu (erbB-2) oncogene. First, a positive correlation between high levels of FAS expression and/or activity and the amplification and/or overexpression of Her-2/neu oncogene exists in human breast cancer cell lines. Second, Her-2/neu overexpression stimulates the activity of FAS gene promoter and ultimately mediates increased endogenous fatty acid biosynthesis, while this Her-2/neu-induced upregulation of breast cancer associated FAS is inhibitable by anti-Her-2/neu antibodies such as trastuzumab (Herceptin(TM)). Third, pharmacological inhibition of FAS activity negatively regulates the expression and tyrosine-kinase activity of Her-2/neu-coded p185(Her 2/neu) oncoprotein. PMID- 16247516 TI - Cardioprotective role of the VIP signaling system. AB - Vasoactive intestinal peptide (VIP) is a 28-amino acid peptide that belongs to a family of structurally related peptide hormones including pituitary adenylate cyclase-activating peptide (PACAP). These hormones are widely distributed in the nervous system, where they act as neurotransmitters. Their biological effects are mediated by specific receptors, VPAC1 and VPAC2, which have comparable affinity for VIP and PACAP, and PAC1, which binds VIP with 1,000-fold lower affinity than PACAP. Both peptides are involved in autonomic regulation of the cardiovascular system, where they exert positive inotropic and chronotropic effects, and cause coronary vasodilatation. Additionally, PACAP inhibits proliferation of cardiac fibroblasts. Several cardiovascular diseases, such as myocardial fibrosis, heart failure, cardiomyopathy and pulmonary hypertension, have been found to be associated with changes in myocardial VIP concentration or with alteration of affinity, density and physiological responsiveness of VIP/PACAP receptors. Application of the peptides or their agonists has beneficial effect in hypertension, heart failure and myocardial fibrosis. Taken together, VIP and PACAP have beneficial effects in various pathological conditions. PMID- 16247518 TI - Molecule of the month: asoprisnil. PMID- 16247517 TI - Filling the gaps: osteogenesis imperfecta. PMID- 16247520 TI - A ruling supreme? Interview by Paul Adams. PMID- 16247519 TI - Latest advances and research in lung cancer. AB - Lung cancer is the biggest killer worldwide among all cancers. In the United States, one third of male and one quarter of female cancer deaths are from lung cancer. In spite of continued research efforts, 5-year survival remains low, at around 15% in the United States and about 10% in Europe. The majority (85%) of cases of lung cancer in men are related to cigarette smoking, and while the proportion of tobacco related cases in women is lower (47%), female deaths from lung cancer are nevertheless catching up with male rates around the world. The 11th World Conference of the International Society for Lung Cancer was held July 3-6, 2005, in Barcelona, Spain, and brought together more than 5,000 delegates to discuss the latest developments and research in the field. PMID- 16247521 TI - Management of ingested foreign bodies. PMID- 16247522 TI - Erythema nodosum and pyoderma gangrenosum in 50 patients with Crohn's disease. AB - Erythema nodosum and pyoderma gangrenosum may occur in Crohn's disease. In the present evaluation of consecutive patients with Crohn's disease spanning more than two decades, erythema nodosum was seen in 45 patients and pyoderma gangrenosum was seen in seven patients. Forty-one of 566 women (7.2%) and nine of 449 men (2.0%) were affected. Of these, 45 (4.4%) had erythema nodosum and seven (0.7%) had pyoderma gangrenosum, including two (0.2%) with both dermatological disorders at different times during their clinical courses. Recurrent erythema nodosum was also detected in nine patients (20%) including eight women, while recurrent pyoderma gangrenosum was seen in two patients (28.6%). There was an age dependent effect on the appearance of erythema nodosum in women, with the highest percentages seen in those younger than 20 years of age. Detection rates for erythema nodosum in women only approached the low men's rates in Crohn's disease at older than 40 years of age. Most patients with these dermatological disorders had colonic disease with or without ileal involvement as well as complex disease, usually with penetrating complications. The present study documents a sex-based and age-dependent effect on the clinical expression of erythema nodosum in Crohn's disease. This suggests that some components of the inflammatory process in Crohn's disease may be modulated by estrogen-mediated events, particularly in adolescents and young adults. PMID- 16247523 TI - Antimicrobial therapy in patients with acute variceal hemorrhage. AB - BACKGROUND: Acute variceal hemorrhage is a serious complication of liver disease and hospital outcome is closely related to infection. Patients with cirrhosis are at greater risk for developing bacterial infection, which is associated with failure to control bleeding and higher rates of hospital mortality. Many clinical practice guidelines endorse antimicrobial prophylaxis as standard of care for cirrhotic patients. OBJECTIVE: The present study was performed to characterize the use of antimicrobial therapy for patients hospitalized with acute variceal hemorrhage. METHODS: Medical records of 98 patients hospitalized with suspected variceal hemorrhage were retrospectively reviewed. RESULTS: One-half of the patients received antimicrobials at any time during their hospital admission, and in very few (24%) could prescribed therapy be considered prophylactic. Seventy seven per cent of patients undergoing endoscopy did not receive an antimicrobial within 24 h of the procedure. Those who received antimicrobial therapy had more severe liver disease (model for end-stage liver disease scores of 19.5 +/- 10 versus 12.9 +/- 8, P< 0.05; Child-Pugh class C 78% versus 65%, not significant) and worse in-hospital outcome (length of stay 17 versus 6.5 days, P<0.05; mortality 15 versus two, P< 0.05). Cephalosporins were the most widely prescribed agents (45%), followed by fluoroquinolone (40%). Regimens ranged in length from single-dose administration to two weeks. CONCLUSIONS: Patients with liver disease admitted with variceal hemorrhage were often not prescribed antimicrobial therapy to reduce the risk of bacterial infection. These results imply that published practice guidelines are not being consistently observed. A large, well-designed study with mortality outcome may be required for clinical guidelines to be successfully implemented in practice. PMID- 16247524 TI - Gastrointestinal complications after cardiopulmonary bypass: sixteen years of experience. AB - BACKGROUND: Gastrointestinal (GI) complications are one of the serious complications of cardiac surgery. Although rarely seen, they cause major morbidity and mortality. The aim of the present study was to retrospectively analyze the risk factors acting on the GI complications seen after cardiac operations performed under cardiopulmonary bypass. METHOD: The present study was designed to retrospectively evaluate 13,544 patients who underwent cardiac surgery under cardiopulmonary bypass, between 1988 and 2004 in the authors' clinic. RESULTS: The overall mortality was 346 (2.55%) of 13,544 patients. GI complications developed in 128 patients (0.94%). Among those, 18 (14.1%) died because of GI complications, the most common of which was bleeding. Mesenteric ischemia had the highest case-fatality rate at 71.4%. Valve surgery, concomitant valve and coronary artery bypass grafting surgery, preoperative chronic renal dysfunction, postoperative acute renal failure, deep sternal infection, prolonged ventilation, need for intra-aortic balloon pump and ejection fraction less than 30% were found to be risk factors acting on GI complications. CONCLUSION: GI complications remain a significant concern after cardiac surgery under cardiopulmonary bypass. Higher-risk patients can be identified and treated prophylactically and in the postoperative period. PMID- 16247525 TI - Has the introduction of laparoscopic Heller myotomy altered the treatment paradigm of achalasia? AB - Although surgical myotomy is well established as the most effective and durable treatment for achalasia, wide acceptance of this procedure as a first-line treatment has been hampered by perceived invasiveness and morbidity. Laparoscopic myotomy has significantly reduced surgical trauma and morbidity while maintaining effectiveness. The effect of laparoscopic myotomy on the treatment pattern for achalasia is not currently known. All patients undergoing surgical myotomy in Quebec from 1997 to 2002 were identified from the Regie de l'assurance maladie du Quebec billing database; previous endoscopic treatment was documented from 1990 to the time of surgery. Patients were divided into two groups (prelaparoscopy and postlaparoscopy) defined by the approximate date when laparoscopic myotomy became generally available in Quebec. A questionnaire examining treatment preference for achalasia was sent to all Quebec gastroenterologists. The number of myotomies performed in Quebec remained stable (prelaparoscopy = 28.7/year; postlaparoscopy = 33/year), but were performed on an older population. The rate of preoperative endoscopic treatment did not differ from prelaparoscopy (29.2%) to postlaparoscopy (23.3%). However, the time interval between the last endoscopy and myotomy diminished significantly. Questionnaire response rate was 41% (60 of 147). Although myotomy was recognized as the most effective treatment (54 of 60), only 22 of 60 gastroenterologists would refer a healthy patient for myotomy as initial treatment. Other choices included dilation (33 of 60), Botulinum toxin (two of 60) or calcium channel blockade (three of 60). Despite a decrease in time interval between endoscopic treatment and surgery, no decrease in the rate of existing endoscopic therapies occurred after laparoscopic myotomy became widely available. The benefits and minimal risks associated with laparoscopic myotomy need to be more effectively communicated by referring physicians. PMID- 16247526 TI - Lamivudine for the treatment of membranous glomerulopathy secondary to chronic Hepatitis B infection. AB - Membranous glomerulopathy is a well-recognized extrahepatic manifestation of chronic hepatitis B virus (HBV) infection. The authors report two cases of HBV related nephrotic syndrome treated with lamivudine. A 46-year-old Chinese man had a hepatitis B e antigen seroconversion along with improvement in his nephrotic syndrome after lamivudine therapy. Two years after treatment was discontinued, a reactivation of HBV was successfully treated again with lamivudine. A 44-year-old Chinese woman, who was intolerant of interferon, was treated with lamivudine for 15 months without a virological response. However, two years after completing lamivudine, her nephrotic syndrome resolved. Implications for the treatment of HBV-related glomerulopathy and a review of the literature are presented. PMID- 16247527 TI - Primary biliary cirrhosis in a patient with Turner syndrome. AB - An increased prevalence of X chromosome monosomy has recently been demonstrated in patients with primary biliary cirrhosis (PBC). Chronic cholestasis of unknown etiology is a common clinical feature in patients with Turner syndrome who reach the fourth and fifth decades of life. A 37-year-old patient with Turner syndrome who presented with clinical and biochemical features of chronic cholestasis is described. Subsequent investigations confirmed the diagnosis of PBC. The patient did not respond to the medical treatment and was referred for liver transplant assessment. The present case may support the importance of X chromosome genes in the development of genetic predisposition to PBC, and emphasizes the necessity for a systematic study of the prevalence of PBC in patients with Turner syndrome. PMID- 16247528 TI - Canadian Cardiovascular Society and Canadian Thoracic Society position statement on pulmonary arterial hypertension. AB - The Canadian Cardiovascular Society and the Canadian Thoracic Society requested a position statement on pulmonary arterial hypertension from leading Canadian experts. The present document is intended to act as an update for the clinician, to provide a template for the initial evaluation of patients, to enable the understanding of current therapeutic paradigms based on approved indications for Canada, to highlight new therapies on the horizon, and to state the positions of the Canadian Cardiovascular Society and the Canadian Thoracic Society on resource management for pulmonary arterial hypertension in Canada. PMID- 16247529 TI - Pediatric interstitial lung disease masquerading as difficult asthma: management dilemmas for rare lung disease in children. AB - Idiopathic nontransplant-related childhood bronchiolitis obliterans is an uncommon disease. Most patients present with chronic recurrent dyspnea, cough and wheezing, which are also features of asthma, by far a much more common condition. The present case study reports on a six-year-old girl who presented to a tertiary care centre with recurrent episodes of respiratory distress on a background of baseline tachypnea, chronic hypoxemia and exertional dyspnea. Her past medical history revealed significant lung disease in infancy, including respiratory syncytial virus bronchiolitis and repaired gastroesophageal reflux. She was treated for 'asthma exacerbations' throughout her early childhood years. Bronchiolitis obliterans was subsequently diagnosed with an open lung biopsy. She did not have sustained improvement with systemic corticosteroids, hydroxychloroquine or clarithromycin. Cardiac catheterization confirmed the presence of secondary pulmonary hypertension. Treatment options remain a dilemma for this patient because there is no known effective treatment for this condition, and the natural history is not well understood. The present case demonstrates the need for careful workup in 'atypical asthma', and the urgent need for further research into the rare lung diseases of childhood. PMID- 16247530 TI - Regional variations in risk factors for asthma in school children. AB - BACKGROUND: The authors have previously reported an increased prevalence of asthma in Estevan, Saskatchewan (21.4%) compared with Swift Current, Saskatchewan (16.2%). OBJECTIVE: To determine the association between asthma and personal and indoor environmental risk factors in these communities. METHODS: A population based cross-sectional study was conducted in January 2000. A questionnaire was distributed to school children in grades 1 to 6 for completion by a parent. Multivariate logistic regression was used to examine associations between various risk factors and physician-diagnosed asthma. RESULTS: Asthma was associated with respiratory allergy (adjusted OR [adjOR]=8.85, 95% CI 6.79 to 11.54), early respiratory illness (adjOR=2.81, 95% CI 1.96 to 4.03) and family history of asthma (adjOR=2.37, 95% CI 1.67 to 3.36). Several environmental factors varied with asthma by town. In Estevan, asthma was associated with home mould or dampness (adjOR=1.82, 95% CI 1.23 to 2.69) and was inversely associated with air conditioning (adjOR=0.56, 95% CI 0.37 to 0.85). The risk of asthma was increased if the child had previous exposure to environmental tobacco smoke from the mother in both communities (Swift Current: OR=1.87, 95% CI 1.06 to 3.30; Estevan: OR=2.00, 95% CI 1.17 to 3.43), and there was an inverse association with current exposure to environmental tobacco smoke from the mother in Estevan (OR=0.64, 95% CI 0.40 to 1.00). When multivariate analyses were stratified by sex, the relationship between home mould or dampness and asthma was most prominent in girls in Estevan. CONCLUSIONS: Despite a similar regional location, different risk factors for asthma were identified in each community. Local environmental factors are important to consider when interpreting findings and planning asthma care. PMID- 16247531 TI - Effects of inhaled fenoterol and positive end-expiratory pressure on the respiratory mechanics of patients with chronic obstructive pulmonary disease. AB - BACKGROUND: During acute ventilatory failure in patients with chronic obstructive pulmonary disease (COPD), applying external positive end-expiratory pressure (PEEPe) will reopen small airways and, thus, may enhance peripheral deposition as well as the physiological effects of inhaled beta-2 agonists. OBJECTIVE: To investigate the efficacy of inhaled fenoterol applied by zero end-expiratory pressure (ZEEPe) or PEEPe. METHODS: Ten patients with COPD who were intubated and mechanically ventilated received fenoterol (10 mg/4 mL) via the ventilator using a jet nebulizer for 30 min on ZEEPe and PEEPe set at 80% of the total PEEP in a random order. The total resistance of the respiratory system (rapid airway occlusion technique), change in end-expiratory lung volume and expiratory flow limitation were assessed before and 5 min, 15 min, 30 min, 60 min and 240 min after fenoterol inhalation. RESULTS: Before inhalation and 60 min after inhalation, the total PEEP, the change in end-expiratory lung volume and the total resistance of the respiratory system were 8+/-3 cmH2O and 6+/-3 cmH2O, 0.61+/-0.34 L and 0.43+/-0.32 L, and 26+/-7 cmH2O/L/s and 23+/-6 cmH2O/L/s, respectively, with ZEEPe, and 9+/-3 cmH2O and 8+/-3 cmH2O (P<0.05 versus ZEEPe), 0.62+/-0.34 L and 0.62+/-0.37 L (P<0.05 versus ZEEPe), and 26+/-9 H2O/L/s and 25+/-9 H2O/L/s, respectively, with PEEPe. Three patients became not flow-limited under the combination of PEEPe and fenoterol. CONCLUSIONS: In patients with COPD, fenoterol combined with PEEPe has opposing effects on respiratory mechanics. First, it does not significantly reduce lung hyperinflation or inspiratory resistances. Second, it allows expiratory flow limitation reversal in some patients. These findings result from the net effect on end-expiratory lung volume of each intervention. This implies that if fenoterol is used in combination with PEEPe, the level of PEEPe should be reassessed during the time course of the drug to prevent any further lung hyperinflation. PMID- 16247532 TI - Hepatic actinomycosis with infiltration of the diaphragm and right lung: a case report. AB - Actinomycosis is an indolent, slowly progressive infection caused by anaerobic or microaerophilic bacteria of the genus Actinomyces. Actinomycosis has a myriad of clinical presentations, inducing both a suppurative and granulomatous inflammatory response. The infection spreads contiguously through anatomical barriers and frequently forms external sinuses. The most common clinical presentations are cervicofacial, thoracic, abdominal and, in females, genital. Classic features include purulent foci surrounded by dense fibrosis that, over time, cross natural boundaries into contiguous structures, with the formation of fistulas and sinus tracts in some cases. Hepatic actinomycosis presents as single or multiple abscesses or masses. Reported here is the unusual occurrence of actinomycosis of the liver involving the diaphragm and right lung. The present case illustrates the difficulties in diagnosing this rare and unrecognized disease. PMID- 16247533 TI - [The impact of L-carnitine on nutritional status in juvenile idiopathic dilated cardiomyopathy]. PMID- 16247534 TI - [Celiac disease: a disease with many faces]. PMID- 16247535 TI - [Birth weight: a cause for concern over both the short and long terms]. PMID- 16247536 TI - [Cow's milk consumption and iron deficiency anemia in children]. AB - OBJECTIVE: To thoroughly investigate the association between the consumption of cow's milk and anemia in childhood. SOURCES OF DATA: The information was gathered from papers catalogued in Lilacs and MEDLINE and published during the last two decades, and also from textbooks and publications by international organizations. SUMMARY OF THE FINDINGS: Iron deficiency anemia is a severe public health problem in developing countries. Breast milk contains adequate iron for full term babies in the first 6 months. Thereafter, an additional iron-rich diet becomes essential. In recognition of the importance of the diet in triggering anemia, this paper discusses the relationship in children between a high intake of cow's milk and iron deficiency anemia. Gastrointestinal and allergic problems may be caused by early introduction of cow's milk or by its substitution for breast milk. Furthermore, cow's milk has decreased iron density and bioavailability, excess protein and minerals, notably calcium, and thus interferes in the absorption of iron from other foods, and is also linked to small intestinal hemorrhage in young children. CONCLUSIONS: The use of cow's milk in lieu of other foods rich in bioavailable iron was shown to be a risk factor for anemia. Exclusive breastfeeding during the first 6 months of life, discretionary weaning only after the 24th month, and a complementary diet rich in iron are highly important to avoid anemia and its consequences. PMID- 16247537 TI - [The role of L-carnitine in nutritional status and echocardiographic parameters in idiopathic dilated cardiomyopathy in children]. AB - OBJECTIVES: Malnutrition is an independent predictor of death in idiopathic dilated cardiomyopathy. An analysis was performed of the impact of L-carnitine supplementation on the nutritional status and echocardiogram parameters of children with idiopathic dilated cardiomyopathy. METHODS: This was an open label cohort of 11 patients who received L-carnitine (100 mg/kg/day) plus the conventional medical treatment, compared with 40 controls, matched for gender and age. The L-carnitine group was weighed 118 times and the controls 264 times. Additionally, the L-carnitine group underwent 65 two-dimensional echocardiograms and the controls 144. Chi-square, Student's t test, Person correlation and ANOVA were calculated with alpha = 0.05. RESULTS: For the L-carnitine group: age at presentation = 3.82 years old, 72.7% (p = 0.033) were females younger than 2 years and 90.9% (p = 0.0001) were in functional classes III and IV. There were no deaths during this period. At presentation, no differences were observed in weight percentile (31.2+/-8.74 vs. 19.6+/-21.2) (p = 0.29) or z score (-0.68+/ 1.05 vs. -1.16+/-0.89) (p = 0.24). Increases were observed in both the percentile (p = 0.026) and z score (p = 0.033) after the introduction of L-carnitine. At presentation, there were no differences in ejection fraction (54.9%+/-3.8 vs. 49.3%+/-6.6) (p = 0.19), but LV mass/BSA were greater in the L-carnitine group (169.12 g/m2+/-26.24 vs. 110.67 g/m2+/-15.62) (p = 0.0005). After the introduction of L-carnitine an increase in ejection fraction (48.3+/-7 to 67.2+/ 7) (p = 0.044) was observed. LV mass/BSA decreased (164.29 g/m2+/-28.14 to 110.88 g/m2+/-28.88), but without significance (p = 0.089) CONCLUSION: In children with idiopathic dilated cardiomyopathy, supplementation of L-carnitine may be helpful for nutritional and echocardiographic improvement. PMID- 16247539 TI - [Does birth weight affect nutritional status at the end of first year of life?]. AB - OBJECTIVE: To evaluate the association between low birth weight and nutritional status at the end of the first year of life. METHODS: This was a nested case control study within a cohort. The study was carried out at maternity hospitals in four cities in the Zona da Mata Meridional in Pernambuco state, Brazil. Newborn infants were recruited during the first 24 hours of life. Their weights were measured at birth and at the end of the first year of life. Household visits were made twice weekly during the first year of life to collect data on breastfeeding and occurrence of diarrhea. In the case-control study, each case (child at nutritional risk) was a child with weight-for-age index < the 10th percentile (n = 117) and each control was a child with weight-for-age index > or = the 10th percentile (n = 411). Hierarchical logistic regression analysis was used to investigate risk factors for nutritional status at 12 months. RESULTS: Low birth weight and living in a household with no latrine were significantly associated with nutritional risk at the end of the first year of life. Children born weighing 1,500 g to 2,499 g had 29 times (95% CI = 9.77-87.49) the chance of being at nutritional risk at 12 months of life than those whose birth weights had been > 3,500 g (p < 0.001). Children living in households without a flush toilet had three times (95% CI = 1.54-6.22) the chance of nutritional risk at 12 months of life in relation to those that had a latrine with a septic tank at home (p = 0.01). CONCLUSION: Low birth weight is an important risk factor of nutritional risk at the end of the first year of life. It is important to adopt strategies for its reduction and prevention. PMID- 16247538 TI - Celiac disease in children and adolescents with Down syndrome. AB - OBJECTIVES: High prevalence rates of celiac disease in patients with Down syndrome have been reported in several countries. However, in Brazil there is no data regarding this association. In this study we report the prevalence of celiac disease in Down syndrome children and adolescents from southern Brazil. METHODS: Seventy-one patients (32 female and 39 male, 2-18 years) from Curitiba, Brazil, were studied. Eighty young people (42 male and 38 female, 2-19 years) were used as controls. All subjects were screened for the IgA-antiendomysium antibody (EmA) and IgA anti-tecidual transglutaminase (anti-tTG). EmA was measured by an immunofluorescence assay using umbilical cord as the substrate and anti-tTG by ELISA with tecidual transglutaminase as the antigen. The total IgA serum level was determined by turbidimetry. RESULTS: Five DS patients (7%) were positive for EmA-IgA, with titers from 1/5 to 1/80 and 14 (17.5%) for anti-tTG (21-340 units). All EmA positive patients also presented anti-tTG antibodies simultaneously. Clinical and histological findings of the intestinal mucosa confirmed celiac disease diagnoses in four patients. The other EmA positive patient was asymptomatic and was not submitted to duodenal biopsy. Patients only positive for anti-tTG presented borderline values (< 25 units) and were asymptomatic. None of the controls were positive for EmA or anti-tTG. No Down syndrome patients or controls presented IgA deficiency. CONCLUSIONS: These data indicates a high prevalence (5.6%) of confirmed celiac disease in Down syndrome patients from southern Brazil. PMID- 16247540 TI - [Nutritional status of Terena indian children from Mato Grosso do Sul, Brazil: follow up of weight and height and current prevalence of anemia]. AB - OBJECTIVE: This study of Terena Indian children less than 10 years old was undertaken in order to compare weight and height in 1995 and 2002 and to evaluate anemia prevalence in 2002. METHODS: This was a cross-sectional study of the Limao Verde and Corrego Seco tribes, located in Aquidauana, Mato Gosso do Sul, Brazil. The study enrolled all 253 children who were less than 10 years old in 1995 and 244 who were under 10 in 2002. Their nutritional status was evaluated comparing weight and height with the NCHS (National Center for Health Statistics) reference values (Epi-Info version 6.2). Hemoglobin was assayed using a portable photometer. RESULTS: The percentages of children with weight for age deficit (z score < -2 standard deviations) were 4 and 2.9% in 1995 and 2002 respectively. Percentages of weight for height deficits were 3.6 and 0.8% respectively. Percentages of height for age deficits were 15 and 11.1% respectively. The median z scores for weight, age and weight, and height showed a statistically significant increase during the period between 1995 and 2002 in infants. Between 24 and 60 months, there were increments in median z scores for weight for age and height for age. There was no statistically significant decrease in any of the anthropometric indicators, with reference to z scores. The prevalence rates of anemia were 86.1% for infants 6 to 24 months old (n = 43), 50.8% for those 24 to 60 months (n = 65) and 40.7% for children aged 60 to 120 months (n = 59). CONCLUSION: Weight and height both increased during period from 1995 to 2002. Anemia was highly prevalent, calling for immediate corrective measures. PMID- 16247541 TI - [Sildenafil in the management of idiopathic pulmonary arterial hypertension in children and adolescents]. AB - OBJECTIVE: This study aims to provide data on the use of oral sildenafil in patients in New York Heart Association functional class III or IV with severe idiopathic pulmonary arterial hypertension unresponsive to conventional therapy. METHOD: In this series, six patients with idiopathic pulmonary arterial hypertension were prospectively treated with 2 to 8 mg of oral sildenafil in four to six doses a day. All patients were submitted to physical examination, electrocardiogram and echocardiogram, chest computed tomography, ventilation and pulmonary perfusion scintigraphy, coagulation studies, and tests for collagen vascular disease, acquired immune deficiency syndrome and schistosomiasis in order to rule out secondary causes of pulmonary arterial hypertension. All patients underwent cardiac catheterization for vasoreactivity tests using nitric oxide, O2 at 100% and oral nifedipine, and a 6 minute walking test was performed in those patients who were considered able to exercise. RESULTS: All patients achieved a good therapeutic response, with improvement by at least one functional class, and presented an increase in systemic arterial oxygen saturation. Five patients showed a decrease in the pulmonary systolic pressure to systemic systolic pressure ratio and improvement in the six-minute walking test. No major side effects were observed at 4 to 36 months of follow-up. One patient had sudden death after sildenafil had been withdrawn by mistake. CONCLUSIONS: These data suggest that sildenafil may be useful in the management of idiopathic pulmonary arterial hypertension. Patients should be advised against the withdrawal of sildenafil without medical supervision. PMID- 16247542 TI - [Endothelial function of normotensive adolescents with no risk factors for arterial hypertension]. AB - OBJECTIVE: To define standards for normal endothelial function in adolescents by high-resolution ultrasound measurement of endothelium-dependent vascular dilatation. METHODS: This was a cross-sectional, descriptive, observational study and part of the thematic project "Clinical Study of Growth, Behavior, Arterial Hypertension, Obesity and Oral Health" (ECCCHOS) that was developed by the Discipline of Nutrition at the Universidade Federal de Sao Paulo. Thirty-one adolescents, eight male and twenty-three female, with no risk factors for systemic arterial hypertension were selected from 1,420 secondary school students. The students were daytime pupils at a school in the southeastern district of the city of Sao Paulo, the capital of Sao Paulo state, located in the Southeast region of Brazil. All results are presented in the form of means with standard deviations and percentiles. RESULTS: For male students, endothelium dependant dilation 90 seconds after the cuff was released was 20.9+/-6.7% [mean +/-1 standard deviation] with a 10th percentile of 12.5 and for females these figures were 18.8+/-12.9% with a 10th percentile of 6.6%. Values for the whole group of subjects were 19.3+/-11.7% and 6.7%, respectively. CONCLUSION: Endothelium-dependant vascular dilation of 6.7%, after 90 seconds, which corresponds to the 10th percentile, can be considered the lower limit of normality for this age group. Knowledge of this limit is important for the diagnosis of endothelium dysfunction that appears before cardiovascular disease. PMID- 16247543 TI - [Conservative management of multicystic dysplastic kidney: clinical course and ultrasound outcome]. AB - OBJECTIVE: The aim of this study was to describe the clinical course and ultrasound outcome of prenatally detected multicystic dysplastic kidney. METHODS: Fifty-three children with unilateral multicystic dysplastic kidney detected by prenatal ultrasound between 1989 and 2004 were included in the analysis. All children were submitted to conservative management with follow-up visits every six months. Follow-up ultrasound examinations were performed at six-month intervals during the first two years of life and yearly thereafter. The following clinical parameters were evaluated: blood pressure, urinary tract infection, renal function, and growth. The following ultrasound parameters were evaluated: involution of multicystic dysplastic kidney and contralateral renal growth. RESULTS: The mean follow-up time was 68 months. Two children presented hypertension during follow-up and five had urinary tract infection (only one with recurrent episodes). There was no malignant degeneration of multicystic dysplastic kidney. A total of 334 ultrasound scans were analyzed. US scan demonstrated involution of the multicystic dysplastic kidney in 48 (90%) cases, including complete involution in nine (17%). The involution rate was faster in the first 30 months of life. There was progressive compensatory renal hypertrophy of the contralateral renal unit; the rate of growth was greater in the first 24 months of life. CONCLUSION: The results of prolonged follow-up of children with conservatively managed multicystic dysplastic kidney suggest that clinical approach is safe, the incidence of complications is small, and that there is a clear tendency for multicystic dysplastic kidney to decrease in size. Our data also suggest that the involution rate of multicystic dysplastic kidney as well as the growth of the contralateral kidney is greater in the first 24 months of life. PMID- 16247544 TI - [The frequency of pharmacological pain relief in university neonatal intensive care units]. AB - OBJECTIVE: To evaluate the use of drugs to relieve procedural pain of newborn infants hospitalized in Neonatal Intensive Care Units (NICU) of university hospitals. METHODS: A prospective cohort study of all newborn infants hospitalized in four NICU during October 2001. The following data were collected: demographic data of patients; clinical morbidity; number of potentially painful procedures and frequency of analgesic administration. Multiple linear regression analysis was performed to study the factors associated with the use of analgesia in this cohort of patients with SPSS 8.0. RESULTS: Ninety-one newborn infants were admitted to the NICU during the study period (1,025 patient-days). Only 25% of the 1,025 patient-days received any systemic analgesia. No specific drug was administered to relieve acute pain during any of the following painful procedures: arterial, venous, capillary and lumbar punctures and tracheal intubation. For chest tube insertion, 100% of newborn infants received specific analgesia. For the insertion of central catheters, 8% of the newborn infants received analgesia. Only nine of the 17 newborn infants that underwent surgical procedures received any dose of analgesics during the postoperative period. Regarding patients who received analgesia, the drug of choice was fentanyl in 93%. The presence of mechanical ventilation increased 6.9 times the chance of the newborn receiving analgesia and the presence of a chest tube increased this chance by 5.0 times. CONCLUSION: It is necessary to train health professionals in order to shorten the lag between scientific knowledge regarding newborn pain and clinical practice. PMID- 16247545 TI - [Prevalence and risk factors for asthma in schoolchildren in southern Brazil]. AB - OBJECTIVE: To study the prevalence and risk factors for asthma in a cohort of 494 children born in 1993 and followed up to the age of six years in Pelotas, state of Rio Grande do Sul, Brazil. METHODS: A standardized and validated asthma questionnaire, based on the International Study of Asthma and Allergies in Childhood (ISAAC), was applied. Other information was also collected about socioeconomic background, genetic, nutritional, gestational and allergic factors, and previous infectious episodes. RESULTS: The prevalence of asthma found in this study was 12.8% (95%CI: 10-15.9%). In the multivariate analysis, risk factors such as non-white skin color (RR = 1.9 95% CI: 1.1-3.3%), family history of asthma (RR = 2.8 95% CI: 1.5-5.1), allergic rhinitis in children (RR = 2.6 95% CI: 1.5-4.4) and maternal smoking during pregnancy (RR = 1.7 95%CI: 1-2.9) were associated with asthma. CONCLUSION: Childhood asthma is highly prevalent in Pelotas, and it is a serious public health problem. Therefore, specific programs should be developed for its control. PMID- 16247546 TI - [Rhinovirus and acute bronchiolitis in young infants]. AB - OBJECTIVE: To determine the prevalence of rhinovirus infection in hospitalized young infants with acute bronchiolitis. METHODS: Hospitalized children with acute bronchiolitis admitted to the Hospital Sao Lucas/PUCRS between May and September 2002 were selected prospectively. Nasopharyngeal samples were assayed for respiratory syncytial virus, parainfluenza, influenza and adenovirus by immunofluorescence. For rhinovirus test a reverse transcription-polymerase chain reaction for picornavirus was used, followed by hybridization with rhinovirus specific probes. RESULTS: Forty-five patients were selected for the study. The median age of the subjects studied was 2 months. Positive samples for respiratory viruses were found in 35/45 (77.8%) subjects and 7/35 (20%) patients had dual infection. Respiratory syncytial virus was detected in 33/35 (94%) cases. Rhinovirus was detected in 6/35 patients (17%). CONCLUSIONS: Rhinovirus was the second most common agent detected in nasal secretions from young infants hospitalized with acute bronchiolitis. PMID- 16247547 TI - [Persistent neonatal hyperbilirubinemia resulting from Gilbert's syndrome in association with RhD hemolytic disease]. AB - OBJECTIVE: To report on an infrequent association of pathologies causing considerable increase in bilirubin production and a significant decrease in its excretion. DESCRIPTION: The third pregnancy of an RhD negative woman. Her first child was normal and delivered to term and did not receive Rhogam. The second pregnancy was problematic due to Rh isoimmunization. She delivered a child to term, who required three exchange transfusions, but died on the 8th day of life. The third child was delivered at term, ORh positive, direct Coombs positive and had stump bilirubin of 6.5 mg/dl and hematocrit at 44%. Five hours after birth the child was jaundiced. Phenobarbital and intensive phototherapy were introduced. Hyperbilirubinemia was soon controlled, but relapsed whenever phototherapy was discontinued. On the 10th day of life the child received a transfusion for significant anemia. As jaundice persisted to the 13th day, associated Gilbert syndrome was considered and DNA sequence analysis was requested. The test demonstrated a mutant homozygote genotype UDPT1A1[TA]7TAA. Phototherapy remained necessary until the 17th day of life and she was discharged from hospital the following day, after bilirubinemia had been controlled. She returned for follow-up and exhibited normal growth and neurological development. COMMENTS: This case demonstrates the significance of increased bilirubin production/decreased bilirubin excretion causing intense hyperbilirubinemias and, in the absence of vigorous treatment, kernicterus. The effectiveness of intense phototherapy has also been demonstrated, reducing the risks of more aggressive treatments such as exchange transfusion. It also provides evidence of the importance of bilirubinemia follow-up until complete resolution. PMID- 16247548 TI - Health-related quality of life in rheumatoid arthritis: therapeutic education plus pharmacological treatment versus pharmacological treatment only. AB - OBJECTIVE: To determine whether therapeutic education added to conventional drug therapy reduced disability and pain in patients with early rheumatoid arthritis (RA). METHODS: Fourty-three patients with RA, 29F/14 M, were included in a randomized, controlled trial and assigned to a control group receiving conventional pharmacological treatment only (n=21), or an intervention group receiving therapeutic education added to conventional pharmacological treatment (n=22). The main outcome variable was self-reported disability on the Stanford health assessment questionnaire (HAQ). RESULTS: At 18 months, patients in the intervention group had less disability (HAQ), pain intensity, number of tender and swollen joints, and patient's and physician's global assessments (p=0.003, 0.031, 0.003, 0.001, 0.014, and 0.004, respectively) compared with baseline, and improvements in disability and number of tender and swollen joints (p=0.024, 0.040, and 0.003, respectively), compared with controls. CONCLUSIONS: Patients receiving pharmacological treatment and therapeutic education had a better evolution than those receiving only pharmacological treatment. PMID- 16247549 TI - PAX9 and TGFB3 are linked to susceptibility to nonsyndromic cleft lip with or without cleft palate in the Japanese: population-based and family-based candidate gene analyses. AB - The prevalence of nonsyndromic cleft lip with or without cleft palate (CL/P) and cleft palate only (CPO) are believed to be higher in the Japanese than in Americans, Europeans or Africans. The purpose of this study was to investigate, in a Japanese population, relationships between CL/P or CPO and seven candidate genes (TGFB3, DLX3, PAX9, CLPTM1, TBX10, PVRL1, TBX22) that showed positive associations in other populations and are expressed in the oral/lip region in developing mice. We first searched for mutations in these genes among 112 CL/P and 16 CPO patients, and found a heterozygous missense mutation (640A > G, S214G) in exon 3 of PAX9 in two sibs with CL/P and their phenotypically normal mother from a Japanese family. A population-based case-control analysis and a family based transmission disequilibrium test (TDT), using single nucleotide polymorphisms (SNPs), and two-SNP haplotypes of the genes, between the 112 CL/P cases with their parents and 192 controls indicated a significant association at one SNP site, IVS1 + 5321, in TGFB3 with a P-value of 0.0016. Population-based haplotyping revealed that the association was most significant for haplotype "A/A" consisting of IVS1 + 5321 and IVS1 - 1572; TDT also gave a P-value of 0.0252 in this haplotype. PMID- 16247550 TI - Dent's disease and prevalence of renal stones in dialysis patients in Northeastern Italy. AB - Dent's disease (DD) involves nephrocalcinosis, urolithiasis, hypercalciuria, LMW proteinuria, and renal failure in various combinations. Males are affected. It is caused by mutations in the chloride channel CLCN5 gene. It has been suggested that DD is underdiagnosed, occurring in less overt forms, apparently without family history. A possible approach to this problem is to search for CLCN5 mutations in patients who may have a high prevalence of mutations: end-stage renal disease (ESRD) patients with previous calcium, struvite, or radio-opaque (CSR) stones. We looked for CLCN5 mutations in 25 males with ESRD-CSR stones selected from all of the patients (1,901 individuals, of which 1,179 were males) of 15 dialysis units in the Veneto region. One DD patient had a new DD mutation (1070 G > T) in exon 7. The new polymorphism IVS11-67 C > T was detected in intron 11 in one patient and one control. We also found 28 females with ESRD and stone history, and seven more males with ESRD and non-CSR stones. The prevalence of stone formers among dialysis patients in our region was 3.2%, much lower than the prevalence observed in older studies. Struvite stones continue to play a major role in causing stone-associated ESRD . PMID- 16247551 TI - Block copolymer adsorption from a homopolymer melt to an amine-terminated surface. AB - Using neutron reflectometry, the adsorption of diblock copolymers from a neutral polystyrene (PS) matrix is studied as a function of substrate type and non adsorbing block degree of polymerization. The block copolymer is poly(deutero styrene)-block-poly(methyl methacrylate) and the substrates are silicon oxide, SiO(x), and SiO(x) functionalized with (3-aminopropyl)triethoxysilane (APTES). We have determined the equilibrium volume fraction-depth profiles for such films, and compared them with volume fraction profiles generated by self-consistent mean field (SCMF) theory and find good agreement between the experimental and theoretical data. SCMF calculations show that the segmental interaction energy between PS matrix chains and APTES is two orders of magnitude stronger than that between PS and SiO(x). PMID- 16247552 TI - Evaluation of perfusion and viability in hypothermic non-beating isolated porcine hearts using cardiac MRI. AB - The donor heart undergoes degradation during hypothermic storage. An assessment of donor heart preservation is typically done with histological or biochemical methods that are not feasible in the clinical setting. We describe a method to study the donor heart using cardiac perfusion MRI that is potentially feasible for clinical use. Standard cardiectomy was performed in the pig model and the hearts were stored in normal saline at 5 degrees C. Imaging was performed by using a rapid gradient-echo sequence (FLASH) with saturation-recovery preparation for T1-weighting in the short axis and horizontal long axis views. Approximately 80 serial images were acquired at a rate of 1/s during administration of 0.006 mmol/ml Gd-DTPA (500 ml, 1 l/min). Signal intensity vs. time curves were generated for each heart and slice imaged and compared to a 0.006 mmol/ml Gd-DTPA reference. H&E stained biopsies of the LV, RV, and septum were also obtained. The mean duration of heart storage (N=10) was 8.8 h (range 4.2-19.2 h). Histologically, no differences were seen in H&E stained biopsies among hearts at different storage times. However cardiac MRI revealed a decrease in perfusion units in each subsequent heart tested after 4.2 h. (R=0.49). Average peak up slope was used as a surrogate measure for flow capacity through the microvasculature and peak contrast enhancement was used as a measurement of viable microvasculature. The 4 h heart had 83% peak contrast enhancement of the reference standard, as compared to 44% for the 19.2 h heart. The decrease in peak enhancement is directly related to the duration of storage time. No correlation of peak up-slope of the intensity curve to storage time was found. This new application of cardiac MRI in the donor heart is applicable to: (1) assessing marginal hearts, (2) evaluating donor heart preservation techniques, and (3) correlating pre- to post-transplant viability. PMID- 16247553 TI - Caffeine production in tobacco plants by simultaneous expression of three coffee N-methyltrasferases and its potential as a pest repellant. AB - Caffeine (1,3,7-trimethylxanthine) is derived from xanthosine through three successive transfers of methyl groups and a single ribose removal in coffee plants. The methyl group transfer is catalyzed by N-zmethyltransferases, xanthosine methyltransferase (XMT), 7-methylxanthine methyltransferase (MXMT) and 3,7-dimethylxanthine methyltransferase (DXMT). We previously cloned three genes encoding each of these N-methyltransferases from coffee plants, and reconstituted the final sequence of the caffeine synthetic pathway in vitro. In the present study, we simultaneously expressed these coffee genes in tobacco plants (Nicotiana tabacum), using a multiple-gene transfer method, and confirmed successful caffeine production up to 5 microg g(-1) fresh weight in leaves of the resulting transgenic plants. Their effects on feeding behavior of tobacco cutworms (Spodoptera litura), which damage a wide range of crops, were then examined. Leaf disc choice test showed that caterpillars selectively fed on the wild-type control materials, or positively avoided the transgenic materials. The results suggest a novel approach to confer self-defense by producing caffeine in planta. A second generation of transgenic crops containing caffeine may save labor and agricultural costs and also mitigate the environmental load of pesticides in future. PMID- 16247554 TI - Male gametic cell-specific histone gH2A gene of Lilium longiflorum: genomic structure and promoter activity in the generative cell. AB - A genomic clone containing the gH2A gene, a histone variant specifically expressed in male gametic cells within the pollen of Lilium longiflorum, was isolated. Sequence analysis revealed that the coding region of the gene is interrupted by one intron, as is the case with the somatic type of plant histone H2A genes, suggesting derivation from the same ancestral gene containing one intron. In addition, a 2.8-kbp fragment of the 5' upstream region of gH2A contained TATA and CAAT boxes, but neither a plant histone-specific regulatory DNA element nor vegetative cell-specific cis-elements were found. A histochemical study of stable transformants demonstrated that the 5' upstream region of the gene can drive gene expression specifically in the generative cell of pollen; no activity was detectable in the vegetative cell or in other reproductive and vegetative tissues of transgenic Nicotiana tabacum. These results strongly suggest that the generative cell can direct specific gene expression, that this expression may be regulated by a putative male gametic factor, and that the gH2A promoter may therefore serve as a useful male gametic cell fate marker in angiosperms. PMID- 16247555 TI - Plant mitochondria contain at least two i-AAA-like complexes. AB - The FtsH proteases, also called AAA proteases, are membrane-bound ATP-dependent metalloproteases. The Arabidopsis genome contains a total of 12 FtsH-like genes. Two of them, AtFtsH4 and AtFtsH11, encode proteins with a high similarity to Yme1p, a subunit of the i-AAA complex in yeast mitochondria. Phylogenetic analysis groups the AtFtsH4, AtFtsH11 and Yme1 proteins together, with AtFtsH4 being the most similar to Yme1. Using immunological method we demonstrate here that AtFtsH4 is an exclusively mitochondrial protein while AtFtsH11 is found in both chloroplasts and mitochondria. AtFtsH4 and AtFtsH11 proteases are integral parts of the inner mitochondrial membrane and expose their catalytic sites towards the intermembrane space, same as yeast i-AAA. Database searches revealed that orthologs of AtFtsH4 and AtFtsH11 are present in both monocotyledonous and dicotyledonous plants. The two plant i-AAA proteases differ significantly in their termini: the FtsH4 proteins have a characteristic alanine stretch at the C terminal end while FtsH11s have long N-terminal extensions. Blue-native gel electrophoresis revealed that AtFtsH4 and AtFtsH11 form at least two complexes with apparent molecular masses of about 1500 kDa. This finding implies that plants, in contrast to fungi and metazoa, have more than one complex with a topology similar to that of yeast i-AAA. PMID- 16247556 TI - Redundant and distinct functions of the ABA response loci ABA-INSENSITIVE(ABI)5 and ABRE-BINDING FACTOR (ABF)3. AB - Abscisic acid-responsive gene expression is regulated by numerous transcription factors, including a subgroup of basic leucine zipper factors that bind to the conserved cis-acting sequences known as ABA-responsive elements. Although one of these factors, ABA-insensitive 5 (ABI5), was identified genetically, the paucity of genetic data for the other family members has left it unclear whether they perform unique functions or act redundantly to ABI5 or each other. To test for potential redundancy with ABI5, we identified the family members with most similar effects and interactions in transient expression systems (ABF3 and ABF1), then characterized loss-of-function lines for those loci. The abf1 and abf3 monogenic mutant lines had at most minimal effects on germination or seed specific gene expression, but the enhanced ABA- and stress-resistance of abf3 abi5 double mutants revealed redundant action of these genes in multiple stress responses of seeds and seedlings. Although ABI5, ABF3, and ABF1 have some overlapping effects, they appear to antagonistically regulate each other's expression at specific stages. Consequently, loss of any one factor may be partially compensated by increased expression of other family members. PMID- 16247557 TI - Photoregulation of the greening process of wheat seedlings grown in red light*. AB - Wheat seedling grown with their shoot bottom exposed to red light (400 micromol m(-2) s(-1)) either with constant illumination or light-dark cycles did not accumulate chlorophyll. This near-etiolation response was manifested by a critical threshold intensity of red light and did not need continuous illumination. The inhibition of the greening process resulted from reduced synthesis of glutamate-1-semialdehyde and consequent reduction in tetrapyrrole precursor 5-aminolevulinic acid. Red light perceived by the shoot bottom down regulated the protein and/or gene expression of enzymes involved in the biosynthesis of tetrapyrroles. The contents of endogenous cytokinins, i.e., isopentenyl-adenosine and dihydrozeatinriboside, were reduced in seedlings grown in red light having their shoot bottom exposed. Application of exogenous cytokinin and its analogue to roots of seedlings grown in red light reversed the down regulation of the greening process. The reversal of red-light-induced near etiolation morphogenesis by far-red (200 micromol m(-2) s(-1)) or blue (25 mumol m(-2) s(-1)) light suggests that it could be a very high red-irradiance response of phytochrome, in the meristematic layers of the shoot bottom, that works in concert with blue light receptor(s). PMID- 16247559 TI - Complete chloroplast genome sequence of Gycine max and comparative analyses with other legume genomes. AB - Lack of complete chloroplast genome sequences is still one of the major limitations to extending chloroplast genetic engineering technology to useful crops. Therefore, we sequenced the soybean chloroplast genome and compared it to the other completely sequenced legumes, Lotus and Medicago. The chloroplast genome of Glycine is 152,218 basepairs (bp) in length, including a pair of inverted repeats of 25,574 bp of identical sequence separated by a small single copy region of 17,895 bp and a large single copy region of 83,175 bp. The genome contains 111 unique genes, and 19 of these are duplicated in the inverted repeat (IR). Comparisons of Glycine, Lotus and Medicago confirm the organization of legume chloroplast genomes based on previous studies. Gene content of the three legumes is nearly identical. The rpl22 gene is missing from all three legumes, and Medicago is missing rps16 and one copy of the IR. Gene order in Glycine, Lotus, and Medicago differs from the usual gene order for angiosperm chloroplast genomes by the presence of a single, large inversion of 51 kilobases (kb). Detailed analyses of repeated sequences indicate that many of the Glycine repeats that are located in the intergenic spacer regions and introns occur in the same location in the other legumes and in Arabidopsis, suggesting that they may play some functional role. The presence of small repeats of psbA and rbcL in legumes that have lost one copy of the IR indicate that this loss has only occurred once during the evolutionary history of legumes. PMID- 16247558 TI - The methylation cycle and its possible functions in barley endosperm development. AB - Barley endosperm development can be subdivided into the pre-storage, intermediate, storage and desiccation phase. Nothing is known about DNA methylation events involved in different endosperm-specific developmental programmes. A complete set of methylation cycle enzyme genes was identified and investigated by mRNA expression analysis. During the pre-storage phase, methionine synthase and S-adenosylmethionine (AdoMet) synthase genes are expressed at high levels, mainly to produce AdoMet, which might be used for methylation processes as indicated by high expression of methyltransferases HvMET1, HvCMT1 and HvDnmt3-1 as well as AdoHcy hydrolase genes. The methyltransferases, core histones and DNA-unwinding ATPases are co-expressed at the mRNA level. On the contrary, storage protein (prolamin) gene expression is repressed due to CpG methylation. Expression of genes responsible for starch biosynthesis is also developmentally regulated but not methylation-dependent. Thus, during pre-storage phase, activity of HvMET1 and HvCMT1 possibly maintains DNA replication and suppresses specific pathways of maturation. Besides, HvDnmt3 1 might be responsible for differentiation-specific de novo methylation. Expression of methyltransferases HvDnmt3-2 and HvCMT2 peaks during the onset of massive starch accumulation. The enzymes are likely responsible for DNA methylation involved in determining plastid division and amyloplast differentiation as concluded from the patterns of co-expressed genes. Levels of AdoMet decarboxylase mRNA, but not methyltransferase- and AdoHcy mRNA, increase at the beginning of desiccation together with methionine synthase and AdoMet synthase levels. This increase may be indicative for utilization of AdoMet in polyamine production protecting aleuron and embryo cell membranes during desiccation. PMID- 16247560 TI - Genomic analysis of the 12-oxo-phytodienoic acid reductase gene family of Zea mays. AB - The 12-oxo-phytodienoic acid reductases (OPRs) are enzymes that catalyze the reduction of double bonds adjacent to an oxo group in alpha,beta-unsaturated aldehydes or ketones. Some of them have very high substrate specificity and are part of the octadecanoid pathway which convert linolenic acid to the phytohormone jasmonic acid (JA). Sequencing and analysis of ESTs and genomic sequences from available private and public databases revealed that the maize genome encodes eight OPR genes. Southern blot analysis and mapping of individual OPR genes to maize chromosomes using oat maize chromosome addition lines provides independent confirmation of this number of OPR genes in maize. A survey of massively parallel signature sequencing (MPSS) assays revealed that transcripts of each OPR gene accumulate differentially in diverse organs of maize plants suggesting distinct biological functions. Similarly, RNA blot analysis revealed that distinct OPR genes are differentially regulated in response to stress hormones, wounding or pathogen infection. ZmOPR1 and/or ZmOPR2 appear to function in defense responses to pathogens because they are transiently induced by salicylic acid (SA), chitooligosaccharides, and by infection with Cochliobolus carbonum, Cochliobolus heterostrophus and Fusarium verticillioides, but not by wounding. In contrast to these two genes, transcript levels of ZmOPR6 and ZmOPR7 and/or ZmOPR8 are highly induced by wounding or treatments with the wound-associated signaling molecules JA, ethylene and abscisic acid. However, accumulation of ZmOPR6 and ZmOPR7/8 mRNAs was not upregulated by SA treatments or by pathogen infection suggesting specific involvement in the wound-induced defense responses. None of the treatments induced transcripts of ZmOPR3, 4, or 5. PMID- 16247561 TI - Functional characterization of a melon alcohol acyl-transferase gene family involved in the biosynthesis of ester volatiles. Identification of the crucial role of a threonine residue for enzyme activity*. AB - Volatile esters, a major class of compounds contributing to the aroma of many fruit, are synthesized by alcohol acyl-transferases (AAT). We demonstrate here that, in Charentais melon (Cucumis melo var. cantalupensis), AAT are encoded by a gene family of at least four members with amino acid identity ranging from 84% (Cm-AAT1/Cm-AAT2) and 58% (Cm-AAT1/Cm-AAT3) to only 22% (Cm-AAT1/Cm-AAT4). All encoded proteins, except Cm-AAT2, were enzymatically active upon expression in yeast and show differential substrate preferences. Cm-AAT1 protein produces a wide range of short and long-chain acyl esters but has strong preference for the formation of E-2-hexenyl acetate and hexyl hexanoate. Cm-AAT3 also accepts a wide range of substrates but with very strong preference for producing benzyl acetate. Cm-AAT4 is almost exclusively devoted to the formation of acetates, with strong preference for cinnamoyl acetate. Site directed mutagenesis demonstrated that the failure of Cm-AAT2 to produce volatile esters is related to the presence of a 268 alanine residue instead of threonine as in all active AAT proteins. Mutating 268 A into 268-T of Cm-AAT2 restored enzyme activity, while mutating 268-T into 268-A abolished activity of Cm-AAT1. Activities of all three proteins measured with the prefered substrates sharply increase during fruit ripening. The expression of all Cm-AAT genes is up-regulated during ripening and inhibited in antisense ACC oxidase melons and in fruit treated with the ethylene antagonist 1 methylcyclopropene (1-MCP), indicating a positive regulation by ethylene. The data presented in this work suggest that the multiplicity of AAT genes accounts for the great diversity of esters formed in melon. PMID- 16247563 TI - The associations between substance use disorders, schizophrenia-spectrum disorders, and Axis IV psychosocial problems. AB - BACKGROUND: Substance abuse among individuals with schizophrenia-spectrum disorders (SSDs) is associated with a range of adverse psychosocial outcomes in the areas of occupational functioning, housing stability, economic independence, access to health care, and involvement with the legal system. The aim of this study was to estimate the effects of substance use disorders (SUDs), SSDS, and dual diagnosis with both disorders on the risk for six important Axis IV psychosocial problems. This was accomplished using a large dataset of patients who are representative of individuals in routine US psychiatric practice. METHOD: Weighted data from the 1999 Study of Psychiatric Patients and Treatments from a practice-based research network of the American Psychiatric Institute for Research and Education were analyzed. Some 615 US psychiatrists provided detailed clinical, psychosocial, and health services information on 1,843 patients, including 285 patients with one or more SUDs without an SSD, 180 patients with a diagnosis of an SSD without substance abuse comorbidity, and 68 dually diagnosed patients. Logistic regression models were used to determine effect estimates (adjusted odds ratios), and corresponding 95% confidence intervals were calculated. RESULTS: After adjusting for sociodemographic variables and for SSD diagnosis, SUD diagnosis was independently associated with increased risk for five of the Axis IV psychosocial problems of interest (occupational problems, housing problems, economic problems, problems with access to health care services, and problems related to interaction with the legal system/crime) when compared to all other psychiatric patients (n=1,310). After adjusting for the sociodemographic variables and for SUD diagnosis, SSD diagnosis (compared to all other psychiatric diagnoses) was associated with Axis IV economic problems, but not with the other five psychosocial problems of interest. The presence of both an SUD and an SSD diagnosis (dual diagnosis) was associated with a greater risk for four of the six Axis IV psychosocial problems studied, compared to the risks associated with either diagnosis alone. Limiting the substance of abuse to alcohol resulted in similar findings. CONCLUSIONS: Although SUDs are associated with increased risk for poor social adjustment, the comorbidity of SUDs and SSDs is associated with greatly compounded psychosocial burdens. These findings, from a large sample of representative US psychiatric patients, demonstrate the ongoing need for improved services and policies for those specially burdened patients with the dual diagnosis of both an SSD and substance abuse or dependence. PMID- 16247562 TI - Endosperm-preferred expression of maize genes as revealed by transcriptome-wide analysis of expressed sequence tags. AB - The transcriptome-wide endosperm-preferred expression of maize genes was addressed by analyzing a large database of expressed sequence tags (ESTs). We generated 30,531 high quality sequence-reads from the 5'-ends of cDNA libraries from maize endosperm harvested at 10, 15, and 20 days after pollination. A further 196,900 maize sequence-reads retrieved from public databases were added to this endosperm collection to generate MAIZEST, a database with tools for data storage and analysis. MAIZEST contains 227,431 ESTs, one third of which represents developing endosperm and the remaining two-thirds represent transcripts from 49 cDNA libraries constructed from different organs and tissues. Assembling the MAIZEST ESTs generated 29,206 putative transcripts, of which a set of 4032 assembled sequences was composed exclusively of sequences derived from endosperm cDNA libraries. After sequence analysis using overlapping parameters, a sub-set of 2403 assembled sequences was functionally annotated and revealed a wide variety of putative new genes involved in endosperm development and metabolism. PMID- 16247564 TI - How does all this effect hernia surgery and surgery in general? PMID- 16247565 TI - Leachability of protein and metals incorporated into aquatic invertebrates: are species and metals-exposure history important? AB - To partially simulate conditions in fish intestinal tracts, we leached six groups of metals-contaminated invertebrates at pH 2 and pH 7, and analyzed the concentrations of four metals (Cd, Cu, Pb, and Zn) and total protein in the leachates. Four of the groups of invertebrates were benthic macroinvertebrates collected from metals-contaminated rivers (the Clark Fork River in Montana and the Coeur d'Alene River in Idaho, USA); the other two groups of invertebrates (one of which was exposed to metals in the laboratory) were laboratory-reared brine shrimp (Artemia sp.). Additionally, we fractionated the pH 2 leachates using size-exclusion chromatography (SEC). Protein content was 1.3 to 1.4x higher in Artemia than in the benthic macroinvertebrates, and leachability of metals and protein differed considerably among several of the groups of invertebrates. In SEC fractions of the pH 2 leachates from both groups of Artemia, Cu and protein co-eluted; however, Cu and protein did not co-elute in SEC fractions of the leachates from any of the benthic macroinvertebrate groups. Although none of the other three metals co-eluted with protein in any of the pH 2 leachates, one or more of the metals co-eluted with lower-molecular-weight molecules in the leachates from all of the groups of invertebrates. These results suggest fundamental differences in metal-binding properties and protein leachability among some invertebrates. Thus, different invertebrates and different histories of metals exposure might lead to different availability of metals and protein to predators. PMID- 16247566 TI - Cadmium accumulation and antioxidative responses in the Sesbania drummondii callus. AB - The effect of cadmium (Cd) on growth, accumulation, and antioxidative response was studied in Sesbania drummondii callus, cultivated on different concentrations of Cd (0-250 microM) for four weeks. Callus growth was comparable to that of the control for concentrations up to 50 microM Cd; however, concentrations higher than 50 microM affected growth. A concentration of 100 microM Cd inhibited growth by 16%, with respect to control. Cd concentration in callus increased with increasing Cd concentrations in the growth medium. Callus accumulated 530 mg Cd kg(-1) of their dry weight at 100 microM Cd concentration. Sesbania callus responded to Cd-induced oxidative stress by modulating antioxidants (glutathione and other non-protein thiols) level and antioxidative enzymes: superoxide dismutase (SOD), ascorbate peroxidase (APX), and glutathione reductase (GR). The content of the glutathione (GSH) and GSH/GSSG ratio first increased up to a concentration of 50 microM Cd and then decreased. The content of other non protein thiols significantly increased with increasing Cd concentrations in the growth medium. The activities of antioxidative enzymes, SOD, APX, and GR, followed the same trends as antioxidants first increasing up to a concentration of 50 microM Cd and then decreasing. These results suggest that antioxidative defense mechanisms play a significant role in Cd detoxification and accumulation in Sesbania drummondii. PMID- 16247567 TI - Mode of action and growth toxicity of arsenic to tilapia Oreochromis mossambicus can be determined bioenergetically. AB - We present a bioenergetics-based approach to analyze the chronic effects and growth toxicity mode of action in tilapia Oreochromis mossambicus exposed to waterborne As and to predict fish growth under different exposure scenarios. 7 day exposure bioassays showed that tilapia accumulate As when exposed to waterborne As. We conducted growth bioassays to assess chronic As toxicity to tilapia. We incorporated a universal ontogenetic growth model with the DEB(tox )theory to explore the mode of action of As toxicity. Our results show that the specific growth rates of exposed tilapia are inversely proportional to As concentrations and are calculated as 0.76% d(-1) in 0 microg mL(-1), 0.57% d(-1) in 1 microg mL(-1), 0.2 % d(-1) in 2 microg mL(-1), and 0.04% d(-1) in 4 microg mL(-1) As, respectively. We showed that the internal threshold concentration did not change significantly with time, demonstrating that the critical body residue approach is applicable for As toxicity assessment. We distinguished between three modes of action of As, including direct effects on growth and indirect effects by way of maintenance and food consumption. Our results support that decreased feeding accounts for the growth decrease in the case of feeding ad libitum. The feeding decrease model also illustrates the growth trajectories of tilapia during the entire whole life span, suggesting that the maximum biomass of tilapia are 1038.75 g in uncontaminated water and 872.97 g in 1 microg mL(-1), 403.06 g in 2 microg mL(-1), and 336.65 g in 4 microg mL(-1) As, respectively. We suggest that considering modes of action in ecotoxicology not only improves our understanding of the toxicities of chemicals, it is also useful in setting up models and avoiding pitfalls in species- and site-specific environmental risk assessment. This proposed framework for tilapia gives preliminary information relevant to aquacultural and ecologic management. PMID- 16247568 TI - Time-course variations of DNA damage and biomarkers of oxidative stress in tilapia (Oreochromis niloticus) exposed to effluents from a swine industry. AB - DNA damage (Comet assay), lipoperoxidation levels (TBARS), and several biomarkers of oxidative stress such as catalase (CAT), glutathione reductase (GR), glutathione peroxidase (GPx), glutathione S-transferase (GST), and contents of reduced (GSH) and total (TG) glutathione were measured in liver and blood (Comet assay) of tilapia (Oreochromis niloticus) exposed for 7, 15, 30 (subchronic exposure), 60, and 90 days (chronic exposure) to two treatment lagoons of a swine processing plant, the first an anaerobic lagoon and the second a final treatment lagoon. After the 15th day, TBARS increased in fish exposed to both lagoons, decreased on the 30th day, and on the 90th day remained similar to controls. Fish exposed subchronically and chronically to both effluents showed consistently greater DNA damage. The CAT and GPx activities showed similar profiles and were induced only during the first week and during the first and second months. GST activity was induced throughout the experimental period. On the other hand, GR activities showed inverted profiles, with progressively decreased activities in the liver of fish exposed to the anaerobic lagoon, and progressively increased activities in fish exposed to the final lagoon. GSH showed higher contents in liver after 60 and 90 days of exposure to the final lagoon. GSSG contents were higher in fish exposed to the final lagoon throughout the experimental period. After 15 days, tilapia exposed to both lagoons showed enhanced total glutathione contents. The hepatic antioxidant system and biomarkers of oxidative stress such as DNA fragmentation and TBARS contents of tilapia exposed to both lagoons presented biphasic profiles. These changes in the antioxidant status also indicate that the industrial treatment is not adequate to avoid damaging environmental effects. PMID- 16247569 TI - Cine magnetic resonance imaging vs high-resolution ultrasonography for detection of adhesions after laparoscopic and open incisional hernia repair: a matched pair pilot analysis. AB - BACKGROUND: Intraabdominal adhesions represent nonspecific complications before or after laparoscopic or open incisional hernia repair. The objective of this matched control pilot study was to display long-term adhesions noninvasively by applying functional cine magnetic resonance (MR) imaging, as compared with applying high-resolution ultrasonography (US). METHODS: The study group, composed of 17 consecutive patients (12 men and 5 women; mean age, 52 years), underwent laparoscopic intraperitoneal onlay mesh repair using expanded polytetrafluoroethylene (ePTFE) mesh. Their mean body mass index was 30 kg/m(2), and the size of the hernia was 95 cm(2). Another group, matched for age, gender, and type of hernia, was subjected to open abdominal wall repair using the preperitoneal sublay technique with a large-pore, low-weight polypropylene mesh. For cine MR imaging (1.5 T), section-by-section dynamic depiction of induced visceral slide throughout the entire abdomen was achieved by applying transverse or sagittal true fast imaging with steady-state precession sequences. The location and type of adhesions were compared with high-resolution ultrasonography using nine segments of the abdominal map. RESULTS: The patients subjected to laparoscopic and open incisional hernia repair were examined 16 and 28 months after surgery. The findings showed functional cine MR imaging as superior to high resolution ultrasonography for assessing the amount of intraabdominal adhesions (n = 53 vs n = 3; p < 0.01). Most frequently, adhesions were seen between small bowel loops and the abdominal wall (n = 22), followed by bowel-to-bowel adhesions (n = 19; p < 0.05). However, adhesions between small bowel loops and the abdominal wall occurred more frequently after open mesh repair (p < 0.05). Furthermore, a strong correlation was observed between patient complaints and findings with cine MR imaging (p < 0.05). Maximum pain correlated significantly with the region of the most distinctive adhesions (p < 0.05). CONCLUSIONS: Functional cine MR imaging represents a reliable noninvasive technique for detecting long-term adhesions after open and laparoscopic incisional hernia repair. The study results suggest that this approach has distinct advantages over high-resolution ultrasonography. PMID- 16247570 TI - Susceptibility of prosthetic biomaterials to infection. PMID- 16247572 TI - Time for evidence-based minimal access surgery training--simulate or sink. PMID- 16247574 TI - Laparoscopic radiofrequency thermal ablation for unusual hepatic tumors: operative indications and outcomes. AB - BACKGROUND: There is increasing experience with laparoscopic radiofrequency ablation for the treatment of patients with hepatic metastasis from colorectal and neuroendocrine cancer and those with hepatocellular cancer. Little is known about the outcomes for patients with other tumor types. METHODS: Between January 1996 and March 2005, 517 patients with 1,500 primary and metastatic liver tumors underwent laparoscopic radiofrequency ablation. Among these, 53 patients (10%) had cancers other than the colorectal, neuroendocrine, or hepatocellular types including sarcoma (n = 18), breast cancer (n = 10), esophagus cancer (n = 4), melanoma (n = 4), lung cancer (n = 3), ovarian cancer (n = 2), pancreas cancer (n = 2), unknown primary cancer (n = 2), cholangiocarcinoma (n = 2), rectal squamous cancer (n = 2), renal cancer (n = 2), papillary thyroid cancer (n = 1), and hemangioendothelioma (n = 1). Unlike the criteria for treatment of the more usual tumor types, these patients had a diagnosis of liver-exclusive disease, as diagnosed by preoperative imaging. They also had failed chemotherapy. RESULTS: The 53 patients underwent ablation of 192 lesions, with 8 patients undergoing repeat treatment. The hospital stay averaged 1 day, and there was no 30-day mortality. Complications included one postoperative hemorrhage, one liver abscess, and one wound infection. Tumors recurred locally for 17% of the lesions over a mean follow-up period of 24 months. The overall median survival was 33 months for the whole series, more than 51 months for breast cancer, and 25 months for sarcoma. CONCLUSION: Laparoscopic radiofrequency ablation can safely and effectively treat hepatic metastasis of these unusual tumor types. The authors believe that this heterogeneous group of patients, selected for their unusual presentation of liver-exclusive disease, may benefit from cytoreduction of their tumor by laparoscopic radiofrequency ablation when other treatment methods have failed. PMID- 16247573 TI - Immediate peritoneal response to bacterial contamination during laparoscopic surgery. AB - BACKGROUND: Several studies have shown that laparoscopic surgery (LS) minimizes surgical trauma and the immune function is better preserved. Another major advantage of LS is the lower incidence of septic complications. However, several in vitro studies have shown that CO(2) severely impairs macrophage physiology. In theory, this would reduce the ability to respond to peritoneal contamination. However, there is some controversy in view of the evidence of a better preserved peritoneal response to sepsis. This study analyzed the early response of the peritoneum to contamination in a CO(2) ambience. METHODS: A total of 192 CD-1 mice were distributed in three groups: group 1, laparotomy (LAP, n = 64); group 2, CO(2) laparoscopy (CO(2)-LC, n = 64); and group 3, wall lift laparoscopy (WL LC, n = 64). Mice in each group were randomized to receive 1 ml of Escherichia coli suspension (1 x 10(4) colony-forming units/ml) or saline. Peritoneal fluid was obtained at 1.5, 3, 6, and 12 h after surgery. Monocyte chemoattractant protein-1 (MCP-1), interleukin-6 (IL-6), and prostaglandin E(2) (PGE(2)) were measured. RESULTS: MCP-1 levels were significantly greater and higher earlier in group 2 (CO(2)-LC) than in group 1 (LAP) (p < 0.007). Simultaneously, the increment in the traction group (WL-LC, group 3) was significantly higher (p < 0.002) than after laparotomy, with no differences in group 2 (CO(2)-LC). When a contamination was added to the laparotomy subgroup, there was a significant increase compared to the group without contamination (p < 0.5). MCP-1 modifications after contamination in the LAP group were statistically significant and appeared later than in the WL-LC (p < 0.002) and CO(2)-LC groups (p < 0.02). For IL-6, the three models presented a significant increase in the noncontaminated groups. This occurred significantly later in the LAP group. Simultaneously, the increase in IL-6 occurred earlier and was significantly higher in the WL-LC group compared to the LAP group (p < 0.003), without differences between CO(2)-LC and wall lift groups. Significant differences between contaminated and noncontaminated subgroups were only observed in the LC CO(2) groups. When contaminated, the traction model sustained a higher and earlier rise in IL-6 levels compared to the LAP and LC-CO(2) groups (p < 0.001). For PGE(2), The three models showed a significant increase in PGE(2) levels in the noncontaminated groups. However, there were no significant differences between them. In the contaminated groups, there was no statistical difference between the groups. CONCLUSION: Despite a transient impairment of the immediate peritoneal response to a septic challenge, the degree of injury with LS is lower than that with open surgery, and abdominal infection can therefore be better controlled. PMID- 16247571 TI - Laparoscopy for abdominal emergencies: evidence-based guidelines of the European Association for Endoscopic Surgery. AB - BACKGROUND: Emergency laparoscopic exploration can be used to identify the causative pathology of acute abdominal pain. Laparoscopic surgery also allows treatment of many intraabdominal disorders. This report was prepared to describe the effectiveness of laparoscopic surgery compared to laparotomy or nonoperative treatment. METHODS: A panel of European experts in abdominal and gynecological surgery was assembled and participated in a consensus conference using Delphi methods. The aim was to develop evidence-based recommendations for the most common diseases that may cause acute abdominal pain. RECOMMENDATIONS: Laparoscopic surgery was found to be clearly superior for patients with a presumable diagnosis of perforated peptic ulcer, acute cholecystitis, appendicitis, or pelvic inflammatory disease. In the emergency setting, laparoscopy is of unclear or limited value if adhesive bowel obstruction, acute diverticulitis, nonbiliary pancreatitis, hernia incarceration, or mesenteric ischemia are suspected. In stable patients with acute abdominal pain, noninvasive diagnostics should be fully exhausted before considering explorative surgery. However, diagnostic laparoscopy may be useful if no diagnosis can be found by conventional diagnostics. More clinical data are needed on the use of laparoscopy after blunt or penetrating trauma of the abdomen. CONCLUSIONS: Due to diagnostic and therapeutic advantages, laparoscopic surgery is useful for the majority of conditions underlying acute abdominal pain, but noninvasive diagnostic aids should be exhausted first. Depending on symptom severity, laparoscopy should be advocated if routine diagnostic procedures have failed to yield results. PMID- 16247575 TI - Day-case endoscopic totally extraperitoneal inguinal hernioplasty versus open Lichtenstein hernioplasty for unilateral primary inguinal hernia in males: a randomized trial. AB - BACKGROUND: Endoscopic totally extraperitoneal inguinal hernioplasty (TEP) is an accepted technique for the repair of recurrent and bilateral inguinal hernia, but its role in the management of unilateral primary inguinal hernia remains controversial. The current randomized trial was undertaken to compare the postoperative and 1-year outcomes of day-case TEP and open Lichtenstein hernioplasty for unilateral primary inguinal hernia in males. METHODS: From January 2002 to January 2004, a total of 200 male patients were randomized to undergo either day-case unilateral TEP or open Lichtenstein hernioplasty under general anesthesia. The primary outcome measures included postoperative pain score, time until return to work, incidence of chronic groin pain, and recurrence rate 1 year after the operation. RESULTS: All TEP procedures were successfully performed without conversion. The mean operation time for TEP (50 +/- 13.2 min) was significantly shorter than for open Lichtenstein hernioplasty (58 +/- 17.6 min) (p < 0.001). The pain score at rest was significantly lower in the TEP group than in the open group on postoperative days 0, 1, 4, 5, and 6. On the average, the patients returned to work 8.6 days after TEP and 14 days after Lichtenstein hernioplasty (p = 0.006). Postoperative recovery and morbidity rates were otherwise comparable between the two groups. The incidence of chronic groin pain 1 year after TEP (9.9%) was significantly lower than after open surgery (21.7%) (p = 0.032). None of the patients in either group showed recurrence at the last follow-up assessment. CONCLUSIONS: Day-case TEP was superior to open Lichtenstein hernioplasty for the repair of unilateral primary inguinal hernia in males. The benefits of day-case TEP included less postoperative pain, a faster return to work, and a lower incidence of chronic groin pain. PMID- 16247576 TI - Laparoscopic repair of perforated duodenal ulcers: the"three-stitch" Graham patch technique. AB - BACKGROUND: "Three-stitch" laparoscopic Graham patch repair (LGPR) for perforated duodenal ulcer enjoyed the same advantage as open Graham patch repair. However, it was not a popular approach because it had problems of suture entanglement and difficult laparoscopic suturing and knotting. The authors describe their technique and results. METHODS: A prospective series from January 2000 to September 2004 was examined. In this study, 35 LGPRs were performed for 32 males and 3 females with a median age of 47 years (range, 18-76 years). RESULTS: No conversion occurred for any of the 35 LGPRs attempted. The median perforation size was 5 mm (3-10 mm), and the median operative time was 86 min (range, 55-163 min). The median time for ambulation was day 2, and the median time for discharge was day 4. Morbidity was 11%, involving one chest infection, one retention of urine, one pelvic collection, and one pyloric stenosis. There was no reoperation, leakage, or mortality. CONCLUSION: The authors' LGPR technique was safe and efficient, and might be the choice for laparoscopic repair of relatively large perforations. PMID- 16247577 TI - Depth of endoscopically placed sutures: an experimental study in a human cadaver model. AB - BACKGROUND: Endoscopic suturing devices offer interesting access for interventional procedures used on the gastrointestinal tract. For the time being, the main indication is endoluminal suturing at the gastroesophageal junction for the management of gastroesophageal reflux disease. There is some evidence that endoluminal endoscopic suturing offers an alternative to the closure of esophageal fistulas and to the fixation of feeding tubes and stents in the near future. A review of the literature found no anatomic data on wall layers stitched by sutures. The aim of this study was to determine the depth of sutures placed endoscopically in the esophagus of a human cadaver model. METHODS: Altogether, 62 sutures were placed in the esophagi of 10 cadavers (complete exenterative cadaver model) at three different suction levels (0.4, 0.6, and 0.8 bar) using the EndoCinch suturing machine. After preparation of the esophagus from its mediastinal bed, all sutures were fixed in formalin and stained with hematoxylin and eosin for histologic examination. RESULTS: No sutures were placed in the mucosa alone. As observed, 1.6% were placed in the submucosa, 4.8% in the circular muscularis propria, and 56.5% in the longitudinal muscularis propria, with 37% placed transmurally. At a suction level of 0.4 bar (0.6, 0.8 bar), 0% (0%, 1.6%) were placed in the submucosa, 3.2% (0%, 1.6%) in the circular muscularis propria, and 11% (25.8%, 12.9%) in the longitudinal muscularis propria, with 12.9% (6.5%, 17.7%) placed transmurally. CONCLUSIONS: This study reports, for the first time, a systematic examination of the depth of sutures placed endoscopically in the esophagus. Most of the sutures were found in the muscular wall of the esophagus at a suction level of 0.6 bar. Also, transmural placements were seen. Reduction of suction pressure may lead to a decrease in transmural sutures. PMID- 16247578 TI - Laparoscopically assisted distal gastrectomy with standard radical lymph node dissection for gastric cancer. AB - BACKGROUND: Laparoscopically assisted distal gastrectomy (LADG) with limited lymph node dissection (D1+alpha) has been used to treat a subset of patients with early gastric cancer. Technical advances have expanded indications for LADG to more advanced gastric cancers. However, little data are available on the feasibility or advantages of LADG with standard radical D2 lymph node dissection for patients with gastric cancer. METHODS: This study reviewed the clinical features of 37 patients who underwent LADG with D2 lymph node dissection for preoperatively diagnosed gastric carcinoma, then compared the results with the features of 31 patients who underwent conventional open distal gastrectomy (ODG) with D2 lymph node dissection. RESULTS: The laparoscopic procedure was not converted to laparotomy in any patient. There was no operative mortality and no serious morbidity among the patients who underwent LADG with D2 lymph node dissection. As compared with the ODG group, the LADG group had less operative blood loss (p < 0.001), earlier recovery of bowel activity (p = 0.012), and a shorter duration of fever after surgery (p = 0.015), despite the longer operation time (p = 0.007). CONCLUSIONS: According to this study, LADG with D2 lymph node dissection is feasible and provides several advantages similar to those of limited lymph node dissection (D1+alpha). Depending on surgeons' technical proficiency, LADG can be used with standard radical lymph node dissection for patients with gastric cancers. PMID- 16247579 TI - Telementoring versus on-site mentoring in virtual reality-based surgical training. AB - BACKGROUND: Telementoring can be an adjunct to surgical training using virtual reality surgical simulation. Telementoring is hypothesized to be as effective as a local mentor for surgical skills training. METHODS: In this study, 20 Romanian medical students trained using a virtual reality surgical simulator (LapSim) with a telementor or local mentor. All the students watched an instructional module at the beginning of the exercise. The telementor, in the United States, interacted by videoconferencing. Before and after training sessions, tool path length and time for task completion were measured. RESULTS: Instructional media and training with mentoring resulted in similar levels of performance between locally mentored and telementored groups. Right- and left-hand path length and time decreased significantly within each group from the initial to the final evaluation (p < 0.05) for most tasks (grasping, cutting, suturing). No significant difference was achieved for clip-applying. CONCLUSIONS: Integration of instructional media with telementoring can be as effective for the development of surgical skills as local mentoring. PMID- 16247580 TI - Early versus delayed-interval laparoscopic cholecystectomy for acute cholecystitis: a metaanalysis. AB - BACKGROUND: Early laparoscopic cholecystectomy has been advocated for the management of acute cholecystitis, but little evidence exists to support the superiority of this approach over delayed-interval operation. The current systematic review was undertaken to compare the outcomes and efficacy between early and delayed-interval laparoscopic cholecystectomy for acute cholecystitis in an evidence-based approach using metaanalytical techniques. METHODS: A search of electronic databases, including MEDLINE and EMBASE, was conducted to identify relevant articles published between January 1988 and June 2004. Only randomized or quasi-randomized prospective clinical trials in the English language comparing the outcomes of early and delayed-interval laparoscopic cholecystectomy for acute cholecystitis were recruited. Both qualitative and quantitative statistical analyses were performed. The effect size of outcome parameters was estimated by odds ratio or weighted mean difference where feasible and appropriate. RESULTS: A total of four clinical trials comprising 504 patients met the inclusion criteria. Failure of conservative treatment requiring emergency cholecystectomy occurred for 43 patients (23%) in the delayed group. Metaanalyses demonstrated a significantly shortened total length of hospital stay in the early group (weighted mean difference, -1.12; 95% confidence interval [CI], -1.42 to -0.99; p < 0.001). Pooled estimates did not show any significant differences between the two approaches in terms of operation time, conversion rate, overall complication rate, incidence of bile leakage, and intraabdominal collection. CONCLUSIONS: The safety and efficacy of early and delayed-interval laparoscopic cholecystectomy for acute cholecystitis were comparable. Because evidence suggested that early laparoscopic cholecystectomy reduced the total length of hospital stay and the risk of readmissions attributable to recurrent acute cholecystitis, it is therefore a more cost-effective approach for the management of acute cholecystitis. PMID- 16247581 TI - Association of systemic and thyroid autoimmune diseases. AB - OBJECTIVE: There are few large cohort studies available on the association of systemic and thyroid autoimmune diseases. In this study, we wished to determine the association of Hashimoto's thyroiditis (HT) and Graves' disease (GD) with systemic autoimmune diseases. METHODS: One thousand five hundred and seventeen patients with systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), systemic sclerosis (SSc), mixed connective tissue disease (MCTD), Sjogren's syndrome (SS) and polymyositis/dermatomyositis (PM/DM) were included in the study. The HT and GD were diagnosed based on thorough clinical evaluation, imaging and fine-needle aspiration cytology (FNAC). The frequency of HT and GD in these diseases was assessed. In addition, 426 patients with HT or GD were assessed and the incidence of SLE, RA, SSc, MCTD, SS and PM/DM among these patients was determined. Prevalence ratios indicating the prevalences of GD or HT among our autoimmune patients in comparison to prevalences of GD or HT in the general population were calculated. RESULTS: Altogether 8.2% of systemic autoimmune patients had either HT or GD. MCTD and SS most frequently overlapped with autoimmune thyroid diseases (24 and 10%, respectively). HT was more common among MCTD, SS and RA patients (21, 7 and 6%, respectively) than GD (2.5, 3 and 1.6%, respectively). The prevalences of HT in SLE, RA, SSc, MCTD, SS and PM/DM were 90-, 160-, 220-, 556-, 176- and 69-fold higher than in the general population, respectively. The prevalences of GD in the same systemic diseases were 68-, 50-, 102-, 76-, 74- and 37-fold higher than in the general population, respectively. Among all thyroid patients, 30% had associated systemic disease. In particular, 51% of HT and only 16% of GD subjects had any of the systemic disorders. MCTD, SS, SLE, RA, SSc and PM/DM were all more common among HT patients (20, 17, 7, 4, 2 and 2%, respectively) than in GD individuals (2, 5, 5, 1, 2 and 1%, respectively). CONCLUSION: Systemic and thyroid autoimmune diseases often overlap with each other. HT and GD may be most common among MCTD, SSc and SS patients. On the other hand, these systemic diseases are often present in HT subjects. Therefore it is clinically important to screen patients with systemic autoimmune diseases for the co-existence of thyroid disorders. PMID- 16247582 TI - First report of leprosy presenting as acute polyarthritis in the setting of type I downgrading lepra reaction. AB - Leprosy is a rare cause of acute polyarthritis. We describe the occurrence of oedema of the hands and feet and acute polyarthritis in the setting of type I (downgrading) lepra reaction in an untreated patient with borderline leprosy. This case report further expands the range of articular manifestations that can occur in leprosy. PMID- 16247583 TI - Ankylosing spondylitis and heart abnormalities: do cardiac conduction disorders, valve regurgitation and diastolic dysfunction occur more often in male patients with diagnosed ankylosing spondylitis for over 15 years than in the normal population? AB - The objective of this study was to determine the rate of selected cardiac pathologies (conduction disorders, valve regurgitation and diastolic dysfunction) in patients with long-standing ankylosing spondylitis (AS) and compare the results with the prevalence in the normal population. A rheumatologic (structured questionnaire interview) and cardiac evaluation (resting electrocardiography and echocardiography) was performed in 100 male subjects with AS and a disease duration of more than 15 years. The rates for conduction disorders, aortic and mitral valve regurgitation and diastolic dysfunction were compared with the corresponding results in the literature among the normal population. In patients with long-standing AS there was no increased rate for valve regurgitation (mitral and aortic valve) and for arrhythmia. Diastolic dysfunction occurred more often in patients with long-standing AS. However, this might be caused by the presence of other cardiovascular risk factors such as age and hypertension. According to these results, a cardiologic evaluation with echocardiography should not be recommended routinely in patients with long-standing AS. To confirm these results, a large prospective study with patients with long-standing AS and with a matched control group should be performed in the future. PMID- 16247584 TI - Evaluation of dynamic postural balance using the Biodex Stability System in rheumatoid arthritis patients. AB - The aim of this study was to investigate dynamic postural balance in patients with rheumatoid arthritis (RA) in relation to the disease characteristics. Seventy-four patients with RA and 42 controls of the same age group were tested using the Biodex Stability System (Biodex Medical Systems, Shirley, NY, USA). Anterior/posterior (AP), medial/lateral (ML), and overall (OA) indices were obtained with bilateral stance at platform stabilities of 2 and 8. Subjects were tested with "eyes open" at all times. At the same time, Disease Activity Score, functional disability [Health Assessment Questionnaire, (HAQ)], and Steinbrocker Functional Class (SFC) were assessed. Both the AP and OA indices in the RA group were significantly higher than in the control group for level 8. For OA index, the results were 2.7+/-0.9 in RA and 2.2+/-0.7 in the control group (p=0.006), and for AP index, the results were 2.1+/-0.7 in RA and 1.7+/-0.6 in the control group (p=0.002). Eleven patients (15.9%) and three controls (7.1%) could not complete the test at level 2. When the patients and controls who completed the test were compared, a significant difference was found only in the ML index. The results were 4.6+/-2.4 in RA and 3.8+/-1.6 in the control group (p=0.047). A positive correlation between HAQ and postural balance for all three stability indices at level 8 was detected. A positive correlation between SFC and postural balance for OA and ML at level 8 was also found. Multiple linear regression analyses revealed age and body mass index (BMI) to be the most important factors influencing postural dynamic balance at both levels in the RA group and in healthy controls. RA has a negative effect on dynamic postural stability. The functional status affects dynamic balance more than disease activity. Age and BMI were the most important factors influencing postural dynamic balance in the RA group and in healthy controls. Level 2 does not appear to be an appropriate level for evaluating postural stability in RA. PMID- 16247586 TI - Dramatical improvement of chemoresistant bone lymphoma with rituximab. AB - Non-Hodgkin's lymphoma of the bone is a very rare disease that accounts for approximately 5% of all extranodal non-Hodgkin's lymphomas and for 7-10% of primary bone tumours. We report the case of a 28-year-old man who, in June 2001, presented with a right humerus showing painful destructive lesions with pathological fractures. Biopsy revealed diffuse, large B-cell non-Hodgkin's lymphoma expressing CD20. The patient received six cycles of conventional chemotherapeutic regimen, including cyclophosphamide, doxorubicin, vincristine and prednisone, and VP-16 (etoposide), ifosfamide and mitoxantrone. His arm pain worsened, and x-rays demonstrated progressive disease. He began a trial of rituximab, 750 mg/week, for 4 weeks. There was improvement in pain after the first infusion. Radiographic studies conducted 3 months after rituximab therapy showed marked improvement in his humerus disease. MRI showed a decrease of tumour volume with residual minor signal abnormalities of the bone marrow. He had no evidence of recurrent lymphoma 24 months later. PMID- 16247585 TI - A study on vascular endothelial growth factor and endothelin-1 in patients with extra-articular involvement of rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is a chronic inflammatory autoimmune disease associated with a wide range of extra-articular manifestations. Recent studies emphasise a key inflammatory role of the endothelial cells, either by overexpression of inflammatory mediators or by the proliferation of new blood vessels, in the disease process leading to the systemic organ involvement. To evaluate the relationship between internal organ manifestations and immunological markers of endothelial activation, serum levels of vascular endothelial growth factor (VEGF) and endothelin-1 (ET-1) were determined by an enzyme-linked immunosorbent assay in 64 RA patients and in 32 healthy controls. In comparison with a control group, higher serum concentrations of VEGF and ET-1 (p<0.001) in RA patients were demonstrated. A comparison between both RA groups with (20 patients) and without systemic involvement (44 patients) showed significantly higher concentrations of VEGF (p<0.05) and ET-1 (p<0.01) in the sera of patients with systemic manifestation. Moreover, a significant positive correlation between VEGF and ET-1 (r=0.475, p<0.001) in RA patients was found. A positive correlation between VEGF and Disease Activity Score (DAS) 28 index (r=0.39, p<0.005) as well as erythrocyte sedimentation rate (ESR) (r=0.564, p<0.0001) and C-reactive protein was found. ET-1 serum level correlated significantly with ESR (r=0.326, p<0.05) and DAS 28 index (r=0.307, p<0.05). These results suggest that the elevated serum levels of VEGF and ET-1 are associated with systemic organ involvement in RA patients and may play a key role in the pathogenesis of extra-articular manifestation of the disease. PMID- 16247587 TI - Dynamic gadolinium-enhanced fat-suppressed T1-weighted MRI (chemical shift selective images) for remitting seronegative symmetrical synovitis with pitting edema. AB - A 75-year-old man suddenly suffered from polyarthralgia and pitting edema in his distal extremities. Laboratory tests revealed inflammation, negative rheumatoid factor, and positive B7 human leukocyte antigen typing. Severe synovitis was observed in dynamic gadolinium-enhanced fat-suppressed (DGEFS) T1-weighted images of his right hand. Our diagnosis was remitting seronegative symmetrical synovitis with pitting edema (RS(3)PE syndrome). He responded dramatically to low-dose corticosteroids, after which his synovitis remarkably improved as observed with DGEFS imaging. DGEFS imaging is useful for distribution of synovitis in patients with RS(3)PE syndrome as well as assessing the efficacy of treatment. PMID- 16247588 TI - Synovial haemangioma of the knee: a case report. AB - Synovial haemangioma is a rare but important cause of knee symptoms, which, when undiagnosed, can lead to significant morbidity. Diagnosis is frequently difficult and delayed. We report on a case of synovial haemangioma, which demonstrates the difficulties inherent in diagnosis and the morbidity associated with diagnostic delay, in a young woman. Magnetic resonance imaging (MRI) is a useful tool for diagnosis, but detection on MRI can also be problematic, as shown by this case, demonstrating the need for greater awareness of this condition by both clinicians and radiologists. Arthroscopy is important in both the diagnosis and treatment of these lesions. PMID- 16247589 TI - Rasburicase treatment in severe tophaceous gout: a novel therapeutic option. AB - We recently encountered a destructive case of tophaceous gout in a 57-year-old patient. Despite perfect therapy compliance, the patient failed in the conventional urate-lowering treatment, accounting for the ongoing urate retention and accumulation with progressive tophaceous bulky disease. Application of an experimental scheme of uricolytical therapy on this patient was able to reduce bulky disease significantly. In modern medicine, potent urate-debulking medication with urate oxidase (uricase) derivatives is at our disposal, and it is a challenge for rheumatologists to install the right strategy including innovative approaches with potent uricolytic therapy on the right patient at the right time. PMID- 16247591 TI - Radiologic features of lumbar spine in ochronosis in late stages. AB - Ochronosis is a rare hereditary disorder of tyrosine metabolism. Severe degenerative arthritis and spondylosis occur in the later stages of this disease. Radiologic examinations may reveal changes considered almost pathognomonic for ochronosis. We present the radiologic features of the lumbar spine in two ochronotic patients who were diagnosed after radiologic examinations in the late stages of the disease. PMID- 16247590 TI - Treatment of therapy-resistant sarcoidosis with adalimumab. AB - A possible role of tumor necrosis factor alpha (TNFalpha) in the pathomechanism of sarcoidosis must be considered in the analysis of this disorder since elevated concentrations of this cytokine have been found. In addition, TNFalpha expression could be demonstrated in sarcoid granulomata [1]. It is well known that TNFalpha plays a crucial role in granulomatous inflammation, e.g., in mycobacterial diseases [2]. Therefore, TNFalpha blockade is a potential approach in the therapy for sarcoidosis. Up to now, various cases of therapy-resistant sarcoidosis treated with anti-TNFalpha (infliximab and etanercept) have been reported [3-8]. Here, we describe successful treatment using adalimumab, a human recombinant immunoglobulin (Ig) G1 anti-TNF monoclonal antibody [9]. PMID- 16247593 TI - Fulvestrant ('Faslodex'): extending the reach of endocrine therapy? AB - An effective option for estrogen receptor-positive breast cancer, endocrine therapy has had a significant role in improving the survival of women with postmenopausal breast cancer for more than 20 years. However, acquired resistance to disease by cancer cells is the repeated cause of relapse for patients. This supplement explores current thinking on resistance to endocrine therapy and, in particular, the potential role for fulvestrant ('Faslodex'), a novel endocrine therapy that possesses a unique mechanism of action. PMID- 16247592 TI - Anger as a moderator of safer sex motivation among low-income urban women. AB - Theoretical models suggest that both HIV knowledge and HIV risk perception inform rational decision making and, thus, predict safer sex motivation and behavior. However, the amount of variance explained by knowledge and risk perception is typically small. In this cross-sectional study, we investigated whether the predictive power of HIV knowledge and HIV risk perception on safer sex motivation is affected by trait anger. We hypothesized that anger may disrupt rational decision making, distorting the effects of both HIV knowledge and risk perception on safer sex intentions. Data from 232 low-income, urban women at risk for HIV infection were used to test a path model with past sexual risk behavior, HIV knowledge, and HIV risk perception as predictors of safer sex intentions. Moderator effects of anger on safer sex intentions were tested by simultaneous group comparisons between high-anger and low-anger women (median split). The theoretically expected "rational pattern" was found among low-anger women only, including (a) a positive effect of knowledge on safer sex intentions, and (b) buffer (inhibitor) effects of HIV knowledge and HIV risk perception on the negative path leading from past risk behavior to safer sex intentions. Among high anger women, an "irrational pattern" emerged, with no effects of HIV knowledge and negative effects of both past risk behavior and HIV risk perception on safer sex intentions. In sum, the results suggest that rational knowledge- and risk based decisions regarding safer sex may be limited to low-anger women. PMID- 16247594 TI - Endocrine therapy--current benefits and limitations. AB - Endocrine therapy is a valuable option for the treatment of postmenopausal women with estrogen receptor (ER)-positive breast cancer due to its demonstrated efficacy and favorable safety profile. Although tamoxifen has been the established treatment for more than 20 years its long-term use is associated with several tolerability concerns and may lead to increased risk of endometrial cancer and thromboembolic complications. In addition, many patients who initially respond to treatment with endocrine agents such as tamoxifen eventually relapse with resistant disease. Sequential use of endocrine therapies is often used in patients as resistance to individual agents develops. Several endocrine approaches have been developed that deprive the tumor of estrogen stimulation, either by directly modulating the ER-signaling pathway or by lowering serum or tumor concentrations of estrogen. In the classic pathway of estrogen signal transduction, the steroid hormone binds to its intracellular ER, triggering a cascade of events that ultimately leads to altered gene transcription. More recently, it has become apparent that ER activation can also occur via estrogen independent receptor activation or by non-nuclear action through cell surface receptors. Consequently, molecular cross-talk exists between the ER and growth factor signaling cascades, which is a key factor in de novo and acquired resistance to endocrine therapy. Inappropriate activation of growth factor signaling can readily promote endocrine therapy failure in breast cancer cells, either by overriding the growth-inhibitory properties of antiestrogenic drugs or by establishment of a new self-propagating autocrine loop that efficiently drives resistant cell growth. Fulvestrant is a new type of ER antagonist with no agonist effects that binds, blocks and causes degradation of the ER. As multiple signaling pathways are involved in the activation of ER, the use of agents such as fulvestrant that directly target the ER and lead to both degradation of the receptor and abrogation of ER signaling may prevent or delay the development of absolute endocrine resistance. In addition, combining antiestrogenic drugs with inhibitors of cell signaling molecules to target both the ER and growth factor signaling pathways is likely to provide a means of delaying endocrine therapy resistance, leading the way to more effective breast cancer treatment. PMID- 16247595 TI - Biological characteristics of the pure antiestrogen fulvestrant: overcoming endocrine resistance. AB - Understanding the underlying mechanisms responsible for endocrine resistance remains a challenge in improving the treatment of breast cancer. The discovery that growth factor and estrogen receptor (ER) signaling pathways interact in endocrine resistant breast cancer has provided a rationale for disrupting these signaling cascades in ER-positive, endocrine-resistant tumors. In postmenopausal women, the ER signaling pathway may be targeted using fulvestrant ('Faslodex'), a new type of ER antagonist with no agonist effects. Fulvestrant binds, blocks and causes degradation of the ER, culminating in complete abrogation of estrogen sensitive gene transcription. This unique mechanism of action may result in a lack of cross-resistance with other endocrine agents. Preclinical studies have confirmed the potential of fulvestrant to inhibit the growth of tamoxifen resistant, as well as tamoxifen-sensitive, human breast cancer cell lines. Clinical studies have demonstrated that fulvestrant is an effective treatment option in postmenopausal women with advanced breast cancer who have progressed on prior endocrine therapy. Furthermore, preclinical studies indicate that combining fulvestrant with growth factor targeted agents, such as the epidermal growth factor receptor (EGFR/HER1) tyrosine kinase inhibitor gefitinib (IRESSA) or the anti-human HER2 monoclonal antibody trastuzumab ('Herceptin'), may result in greater anti-tumor activity than either agent alone. A range of clinical trials are now ongoing to determine whether the combination of growth factor-targeting agents with fulvestrant will delay the onset of endocrine resistance and so provide new strategy for women with hormone receptor-positive advanced breast cancer. PMID- 16247596 TI - Life following aromatase inhibitors--where now for endocrine sequencing? AB - The third-generation non-steroidal aromatase inhibitors (AIs) are challenging tamoxifen as treatments of choice for early and advanced breast cancer in postmenopausal women with estrogen receptor (ER)-positive disease. However, patients who initially respond to AIs eventually develop resistance to treatment and experience disease progression. To establish the optimal endocrine therapy following AI resistance, it is essential to understand the mechanisms that contribute to the loss of response. Data from in vitro models have suggested that acquired AI resistance is due to enhanced sensitization to low estrogen levels during long-term estrogen deprivation (LTED). Cross-talk between the ER and various growth-factor-receptor signaling pathways, including human epidermal growth factor receptor 2, and the insulin-like growth factor pathway, may also be implicated. Therefore, endocrine therapies that abolish estrogen signaling via removal of the ER could be effective in patients with AI-resistant disease. Fulvestrant ('Faslodex') is a new ER antagonist with no agonist effects that binds, blocks and degrades the ER. Due to its unique mode of action and lack of cross-resistance with existing treatments, fulvestrant is an effective therapeutic agent for use in sequential endocrine regimens. Fulvestrant has established efficacy in tamoxifen-resistant disease and there is a growing body of evidence demonstrating its efficacy in patients with AI-resistant disease. In preclinical models, MCF-7 cells undergoing LTED are refractory to tamoxifen but sensitive to fulvestrant, suggesting fulvestrant is a more appropriate choice following AI resistance. The steroidal AI, exemestane is also an option in non steroidal AI-resistant disease. Clinical trials are underway to compare fulvestrant with exemestane as an appropriate therapy following the onset of AI resistance. PMID- 16247599 TI - Inhibition of endothelial nitric oxide synthase activity and suppression of endothelium-dependent vasorelaxation by 1,2-naphthoquinone, a component of diesel exhaust particles. AB - Diesel exhaust particles contain redox-active quinones, such as 9,10 phenanthraquinone (9,10-PQ) and 1,2-naphthoquinone (1,2-NQ), which act as potent electron acceptors, thereby altering electron transfer tgljoint @/Set Line Joint Styleon proteins. We have previously found that 9,10-PQ inhibits constitutive nitric oxide synthase (NOS) activity, by shunting electrons away from NADPH on the cytochrome P450 reductase domain of NOS, and thus suppresses acetylcholine (Ach)-induced vasorelaxation in the aortic ring. However, the effect of 1,2-NQ on endothelial NOS (eNOS) activity is still poorly understood. With the membrane fraction of cultured bovine aortic endothelial cells, we found that 1,2-NQ was a potent inhibitor of eNOS with an IC50 value of 1.4 microM, whereas trans-1,2 dihydroxy-1,2-dihydronaphthalene (1,2-DDN), a redox-negative naphthalene analog of 1,2-NQ, did not show such an inhibitory action. Although 1,2-DDN (5 microM) did not affect Ach-mediated vasorelaxation, 1,2-NQ caused a significant suppression of Ach-induced endothelium-dependent vasorelaxation in the aortic ring. However, 1,2-NQ did not affect sodium nitroprusside-induced endothelium independent vasorelaxation. These results suggest that 1,2-NQ is an environmental quinone that inhibits eNOS activity, thereby disrupting NO-dependent vascular tone. PMID- 16247597 TI - Myocardial infarction in diabetic and non-diabetic persons with and without prior myocardial infarction: the FINAMI Study. AB - AIMS/HYPOTHESIS: We compared the risk of acute coronary events in diabetic and non-diabetic persons with and without prior myocardial infarction (MI), stratified by age and sex. METHODS: A Finnish MI-register study known as FINAMI recorded incident MIs and coronary deaths (n=6988) among people aged 45 to 74 years in four areas of Finland between 1993 and 2002. The population-based FINRISK surveys were used to estimate the numbers of persons with prior diabetes and prior MI in the population. RESULTS: Persons with diabetes but no prior MI and persons with prior MI but no diabetes had a markedly greater risk of a coronary event than persons without diabetes and without prior MI. The rate of recurrent MI among non-diabetic men with prior MI was higher than the incidence of first MI among diabetic men aged 45 to 54 years. The rate ratio was 2.14 (95% CI 1.40-3.27) among men aged 50. Among elderly men, diabetes conferred a higher risk than prior MI. Diabetic women had a similar risk of suffering a first MI as non-diabetic women with a prior MI had for suffering a recurrent MI. CONCLUSIONS/INTERPRETATION: Both persons with diabetes but no prior MI, and persons with a prior MI but no diabetes are high-risk individuals. Among men, a prior MI conferred a higher risk of a coronary event than diabetes in the 45-54 year age group, but the situation was reversed in the elderly. Among diabetic women, the risk of suffering a first MI was similar to the risk that non-diabetic women with prior MI had of suffering a recurrent MI. PMID- 16247600 TI - Identification and in silico analysis of a new group of double-histone fold containing proteins. AB - The double-histone fold is a rare protein fold in which two consecutive regions characterized by the typical structure of histones assemble together, thus giving a histone pseudodimer. Previously, this fold was found in a few prokaryotic histones and in the regulatory region of guanine-nucleotide exchange factors of the Sos family. Standard methods of sequence comparison did not allow us to find new proteins containing a histone pseudodimer, as previously reported (Sondermann et al. 2003). However, a deeper investigation of protein sequences showed that the two histone folds included in Sos proteins share significant sequence similarity with nucleosomal histones. On the basis of this observation, we applied a specific strategy of sequence-homology search, which led to the identification of a new group of histone pseudodimers in Cca3 and proteins similar to Cca3 (Cca3S). A homology model of the histone pseudodimer included in rat Cca3 was constructed. A subsequent structure-function relationship study revealed that the histone pseudodimers included in Cca3 and Cca3S proteins, but not those present in Sos proteins, could retain the ability of mediating protein DNA interactions, and could consequently act as DNA-binding modules. PMID- 16247602 TI - Ab initio construction of all-atom loop conformations. AB - In this study, a new ab initio method named CLOOP has been developed to build all atom loop conformations. In this method, a loop main-chain conformation is generated by sampling main-chain dihedral angles from a restrained varphi/psi set, and the side-chain conformations are built randomly. The CHARMM all-atom force field was used to evaluate the loop conformations. Soft core potentials were used to treat the non-bond interactions, and a designed energy-minimization technique was used to close and optimize the loop conformations. It is shown that the two strategies improve the computational efficiency and the loop-closure rate substantially compared to normal minimization methods. CLOOP was used to construct the conformations of 4-, 8-, and 12-residue loops in Fiser's test set. The average main-chain root-mean-square deviations obtained in 1,000 trials for the 10 different loops of each size are 0.33, 1.27, and 2.77 A, respectively. CLOOP can build all-atom loop conformations with a sampling accuracy comparable with previous loop main-chain construction algorithms. [Figure: see text]. PMID- 16247601 TI - Molecular dynamics simulations of bovine rhodopsin: influence of protonation states and different membrane-mimicking environments. AB - G-protein coupled receptors (GPCRs) are a protein family of outstanding pharmaceutical interest. GPCR homology models, based on the crystal structure of bovine rhodopsin, have been shown to be valuable tools in the drug-design process. The initial model is often refined by molecular dynamics (MD) simulations, a procedure that has been recently discussed controversially. We therefore analyzed MD simulations of bovine rhodopsin in order to identify contacts that could serve as constraints in the simulation of homology models. Additionally, the effect of an N-terminal truncation, the nature of the membrane mimic, the influence of varying protonation states of buried residues and the importance of internal water molecules was analyzed. All simulations were carried out using the program-package GROMACS. While N-terminal truncation negatively influenced the overall protein stability, a stable simulation was possible in both solvent environments. As regards the protonation state of titratable sites, the experimental data could be reproduced by the program UHBD (University of Houston Brownian Dynamics), suggesting its application for studying homology models of GPCRs. A high flexibility was observed for internal water molecules at some sites. Finally, interhelical hydrogen-bonding interactions could be derived, which can now serve as constraints in the simulations of GPCR homology models. PMID- 16247603 TI - Effect of diazepam on the efficacy of dual-phase FDG PET imaging. PMID- 16247604 TI - (18)F-FDG PET imaging of rheumatoid knee synovitis correlates with dynamic magnetic resonance and sonographic assessments as well as with the serum level of metalloproteinase-3. AB - PURPOSE: The aim of this study was to assess rheumatoid arthritis (RA) synovitis with positron emission tomography (PET) and( 18)F-fluorodeoxyglucose ((18)F-FDG) in comparison with dynamic magnetic resonance imaging (MRI) and ultrasonography (US). METHODS: Sixteen knees in 16 patients with active RA were assessed with PET, MRI and US at baseline and 4 weeks after initiation of anti-TNF-alpha treatment. All studies were performed within 4 days. Visual and semi-quantitative (standardised uptake value, SUV) analyses of the synovial uptake of FDG were performed. The dynamic enhancement rate and the static enhancement were measured after i.v. gadolinium injection and the synovial thickness was measured in the medial, lateral patellar and suprapatellar recesses by US. Serum levels of C reactive protein (CRP) and metalloproteinase-3 (MMP-3) were also measured. RESULTS: PET was positive in 69% of knees while MRI and US were positive in 69% and 75%. Positivity on one imaging technique was strongly associated with positivity on the other two. PET-positive knees exhibited significantly higher SUVs, higher MRI parameters and greater synovial thickness compared with PET negative knees, whereas serum CRP and MMP-3 levels were not significantly different. SUVs were significantly correlated with all MRI parameters, with synovial thickness and with serum CRP and MMP-3 levels at baseline. Changes in SUVs after 4 weeks were also correlated with changes in MRI parameters and in serum CRP and MMP-3 levels, but not with changes in synovial thickness. CONCLUSION: (18)F-FDG PET is a unique imaging technique for assessing the metabolic activity of synovitis. The PET findings are correlated with MRI and US assessments of the pannus in RA, as well as with the classical serum parameter of inflammation, CRP, and the synovium-derived parameter, serum MMP-3. Further studies are warranted to establish the place of metabolic imaging of synovitis in RA. PMID- 16247606 TI - The evolution of the immunoglobulin heavy chain variable region (IgVH) in Leporids: an unusual case of transspecies polymorphism. AB - In domestic rabbit (Oryctolagus cuniculus), three serological types have been distinguished at the variable domain of the antibody H chain, the so-called V(H) a allotypes a1, a2, and a3. They correspond to highly divergent allelic lineages of the V(H) 1 gene, which is the gene rabbit utilizes in more than 80% of VDJ rearrangements. The sharing of serological V(H) a markers between rabbit and snowshoe hare (Lepus americanus) has suggested that the large genetic distances between rabbit V(H) 1 alleles (9-14% nucleotide differences) can be explained by unusually long lineage persistence times (transspecies polymorphism). Because this interpretation of the serological data is uncertain, we have determined the nucleotide sequences of V(H) genes expressed in specimens of Lepus species. Two sequence groups were distinguished, one of which occurred only in hare specimen displaying serological motifs of the rabbit V(H) a-a2 allotype. Sequences of this group are part of a monophyletic cluster containing the V(H) 1 sequences of the rabbit a2 allotype. The fact that this "transspecies a2 cluster" did not include genes of other rabbit V(H) a allotypes (a1, a3, and a4) is incompatible with the existence of a common V(H) a ancestor gene within the species, and suggests that the divergence of the V(H) a lineages preceded the Lepus vs Oryctolagus split. The sequence data are furthermore compatible with the hypothesis that the V(H)a polymorphism can be two times older than the divergence time between the Lepus and Oryctolagus lineages, which was estimated at 16-24 million years. PMID- 16247607 TI - Inhibition of erythrocyte "apoptosis" by catecholamines. AB - Osmotic shock, oxidative stress and Cl- removal activate a non-selective Ca2+ permeable cation conductance in human erythrocytes. The entry of Ca2+ leads to activation of a scramblase with subsequent exposure of phosphatidylserine at the cell surface. Phosphatidylserine mediates binding to phosphatidylserine receptors on macrophages which engulf and degrade phosphatidylserine exposing cells. Moreover, phosphatidylserine exposure may lead to adherence of erythrocytes to the vascular wall. In the present study, we explored whether activation of the non-selective cation conductance and subsequent phosphatidylserine exposure might be influenced by catecholamines. Phosphatidylserine exposure has been determined by FITC-annexin V binding while cell volume was estimated from forward scatter in FACS analysis. Removal of Cl- enhanced annexin binding and decreased forward scatter, an effect significantly blunted by the beta agonist isoproterenol (IC50 approx. 1 microM). Fluo-3 fluorescence measurements revealed an increase of cytosolic Ca2+ activity following Cl- removal, an effect again significantly blunted by isoproterenol exposure (10 microM). Whole-cell patch-clamp experiments performed in Cl- free bath solution indeed disclosed a time-dependent inactivation of a non-selective cation conductance following isoproterenol exposure (10 microM). Phenylephrine (IC50<10 microM), dobutamine (IC50 approx. 1 microM) and dopamine (IC50 approx. 3 microM) similarly inhibited the effect of Cl removal on annexin binding and forward scatter. In conclusion, several catecholamines inhibit the Cl- removal-activated Ca2+ entry into erythrocytes, thus preventing increase of cytosolic Ca2+ activity, subsequent cell shrinkage and activation of erythrocyte scramblase. The catecholamines thus counteract erythrocyte phosphatidylserine exposure and subsequent clearance of erythrocytes from circulating blood. PMID- 16247609 TI - Analysis of sequence variations in the promoter region of the human tissue factor pathway inhibitor 2 gene in apoplectic patients and blood donors. AB - Tissue factor pathway inhibitor 2 (TFPI-2) is a Kunitz-type serine protease inhibitor with homology to TFPI-1, an important regulator of the extrinsic pathway of blood coagulation. Recent studies have focused on TFPI-2 and its implications for atherosclerosis. The promoter region and the exons of the human TFPI-2 gene were screened for sequence variations in 41 apoplectic patients and 140 blood donors with no history of ischemic stroke. The sequence variations 567T>C, -546T>C, -353A>G, -161G>C, -167G>A, -47C>A, and -18C>A, which are located in the TFPI-2 promoter, were discovered in both cohorts with allelic frequencies ranging from 0.3 to 2.4%. The influence of these sequence variations on the transcriptional activity of the TFPI-2 gene was investigated in HEK-293 cells using a promoter test system. A wild-type TFPI-2 promoter fragment 716 bp upstream of the translation start site was cloned into a secreted alkaline phosphatase expression vector, and the sequence variations were introduced by site-directed mutagenesis. Interestingly, the promoter activity of the tested mutants was reduced by 1.3- to 2.8-fold compared to that of wild-type control. The variation -18C>A, where a putative binding site of the transcription factor Sp-1 is located, had the strongest effect on transcriptional activity. In conclusion, our present study shows that the transcription of TFPI-2 is changed by single nucleotide polymorphisms and that the sequence variations in transcription factor binding sites of the TFPI-2 promoter may influence the regulation of this gene. PMID- 16247608 TI - Occurrence of bicuculline-, NMDA- and kainic acid-induced seizures in prenatally methamphetamine-exposed adult male rats. AB - Stimulant drugs are often associated with increased seizure susceptibility. Inhibitory gamma-aminobutyric acid (GABA) and excitatory N-methyl-D-aspartate (NMDA) systems play an important role in the effect of stimulants on epileptic seizures. No studies investigating the effect of prenatal methamphetamine (MA) exposure on seizures are available. In this study, bicuculline (GABAA receptor antagonist), NMDA (NMDA receptor agonist) and kainic acid (non-NMDA receptor agonist) were used to induce seizures in adult male rats. Three groups of animals were tested in each seizure test: prenatally MA- (5 mg/kg) exposed, prenatally saline-exposed, and absolute controls without any prenatal exposure. In bicuculline-induced seizures, the latency to onset of tonic-clonic seizures was shorter in MA-exposed rats than in controls, but it did not differ from saline exposed rats. There were no differences in clonic seizure onset between groups. In NMDA-induced seizures, the latency to onset of clonic-tonic seizures was shorter in prenatally MA-exposed rats than in controls; however, the latency to onset of saline-exposed animals did not differ from either MA-exposed or from control rats. There were no differences in seizure susceptibility in kainic acid induced clonic seizures. There were no differences in seizure incidences or stereotypical behavior in any seizure model. The question remains as to how much the present data demonstrate the effect of prenatal drug exposure on seizure susceptibility per se, and how much they may be explained by the effect of prenatal stress or by other mechanism(s). PMID- 16247610 TI - Comparative efficacy of two regimens in syndromic management of lower genital infections. AB - AIM: The aim of this study was to compare the efficacy and safety of two combination regimens in the syndromic management of lower genital infection. Seventy-two non-pregnant women presenting with symptoms of lower genital infection diagnosed as vaginitis on clinical examination and lacking obvious upper genital infection were enrolled to one of the two treatment regimens as a syndromic treatment. No investigations were performed to cut the cost and to avoid the loss of patients on follow-up. Thirty-seven women (group I) were prescribed a course of clotrimazole (Imidil, Lyka) 100 mg vaginal pessaries for 6 days. Along with their partners they were prescribed 2 gm secnidazole (Secnil forte) and 150 mg fluconazole (Syscan) as a single therapy. Thirty-five women (group II) were prescribed vaginal clotrimazole as mentioned above. A combination kit containing 150 mg fluconazole, 2 gm secnidazole and 1 gm azithromycin (FAS-3 kit, Lyka) was also prescribed to both partners with the advice to take azithromycin on empty stomach, and the other three tablets after food. RESULTS: All women in both groups were seen after 1 week for relief of symptoms and after 1 month for any recurrence. Mean parity was 2.7 and 3.0, respectively. The total symptomatic relief was observed in 67.6 and 94.3% cases, partial relief in 27 and 5.7% cases and no relief was observed in 5.4% and nil cases, respectively, in the two groups. Recurrence was seen in two and nil cases, respectively, in the two groups. Most women tolerated both the treatments well with no major side effect in any case. Treatment cost was higher in group II (Rupees 120) than in group I (Rupees 65). CONCLUSIONS: Both combination kits with local clotrimazole were reasonably effective and safe in the syndromic approach for lower genital infections. The combination kit with azithromycin, secnidazole and fluconazole was more effective with better symptomatic relief and less recurrence rate and may be routinely recommended in all cases of lower genital infection as a cost effective, safe and effective strategy. PMID- 16247611 TI - Effects of agitation on the microalgae Phaeodactylum tricornutum and Porphyridium cruentum. AB - The effect of mechanical agitation on the microalgae Phaeodactylum tricornutum and Porphyridium cruentum was investigated in aerated continuous cultures with and without the added shear protectant Pluronic F68. Damage to cells was quantified through a decrease in the steady state concentration of the biomass in the photobioreactor. For a given aeration rate, the steady state biomass concentration rose with increasing rate of mechanical agitation until an upper limit on agitation speed was reached. This maximum tolerable agitation speed depended on the microalgal species. Further increase in agitation speed caused a decline in the steady state concentration of the biomass. An impeller tip speed of >1.56 m s(-1) damaged P. tricornutum in aerated culture. In contrast, the damage threshold tip speed for P. cruentum was between 2.45 and 2.89 m s(-1). Mechanical agitation was not the direct cause of cell damage. Damage occurred because of the rupture of small gas bubbles at the surface of the culture, but mechanical agitation was instrumental in generating the bubbles that ultimately damaged the cells. Pluronic F68 protected the cells against damage and increased the steady state concentration of the biomass relative to operation without the additive. The protective effect of Pluronic was concentration-dependent over the concentration range of 0.01-0.10% w/v. PMID- 16247612 TI - Green fluorescent protein as an efficient selection marker for Agrobacterium rhizogenes mediated carrot transformation. AB - Agrobacterium rhizogenes mediated transformation combined with a visual selection for green fluorescent protein (GFP) has been applied effectively in carrot (Daucus carota L.) transformation. Carrot root discs were inoculated with A4, A4T, LBA1334 and LBA9402 strains, all bearing gfp gene in pBIN-m-gfp5-ER. The results indicate that transformed adventitious roots can be visually selected solely based on GFP fluorescence with a very high accuracy. The method requires no selection agents like antibiotics or herbicides and enables a reduction of labour and time necessary for tissue culture. Moreover, individual transformants can be easily excised from the host tissue and cultured separately. All of the 12 used carrot cultivars produced transformed adventitious roots and the frequency of discs producing GFP expressing adventitious roots varied from 13 to 85%. The highest transformation rate was found for A4T and LBA1334 strains possessing chromosomal background of A. tumefaciens C58. The results encourage that visual selection of transformed, fluorescing adventitious roots can be highly effective and applied routinely for the production of carrot transgenic plants. PMID- 16247613 TI - Low light and low ammonium are key factors for guayule leaf tissue shoot organogenesis and transformation. AB - A new method has been developed for guayule tissue culture and transformation. Guayule leaf explants have a poor survival rate when placed on normal MS medium and under normal culture room light conditions. Low light and low ammonium treatment greatly improved shoot organogenesis and transformation from leaf tissues. Using this method, a 35S promoter driven BAR gene and an ubiquitin-3 promoter driven GUS gene (with intron) have been successfully introduced into guayule. These transgenic guayule plants were resistant to the herbicide ammonium glufosinate and were positive to GUS staining. Molecular analysis showed the expected band and signal in all GUS positive transformants. The transformation efficiency with glufosinate selection ranged from 3 to 6%. Transformation with a pBIN19-based plasmid containing a NPTII gene and then selection with kanamycin also works well using this method. The ratio of kanamycin-resistant calli to total starting explants reached 50% in some experiments. PMID- 16247615 TI - Efficacy of micafungin for the treatment of candidemia. PMID- 16247617 TI - Pediatric angiography: what is the future? PMID- 16247616 TI - Krabbe disease: unusual MRI findings. AB - We present the MRI findings in a case of infantile-onset Krabbe disease. Enlargement of the intracranial optic nerves and cervical cord were detected in addition to more typical changes in the cerebral white matter and thalami. We also review the proton MR spectroscopic findings in Krabbe disease. PMID- 16247614 TI - The microarchitecture of DNA replication domains. AB - Most DNA synthesis in HeLa cell nucleus is concentrated in discrete foci. These synthetic sites can be identified by electron microscopy after allowing permeabilized cells to elongate nascent DNA in the presence of biotin-dUTP. Biotin incorporated into nascent DNA can be then immunolabeled with gold particles. Two types of DNA synthetic sites/replication factories can be distinguished at ultrastructural level: (1) electron-dense structures- replication bodies (RB), and (2) focal replication sites with no distinct underlying structure--replication foci (RF). The protein composition of these synthetic sites was studied using double immunogold labeling. We have found that both structures contain (a) proteins involved in DNA replication (DNA polymerase alpha, PCNA), (b) regulators of the cell cycle (cyclin A, cdk2), and (c) RNA processing components like Sm and SS-B/La auto antigens, p80-coilin, hnRNPs A1 and C1/C2. However, at least four regulatory and structural proteins (Cdk1, cyclin B1, PML and lamin B1) differ in their presence in RB and RF. Moreover, in contrast to RF, RB have structural organization. For example, while DNA polymerase alpha, PCNA and hnRNP A1 were diffusely spread throughout RB, hnRNP C1/C2 was found only at the very outside. Surprisingly, RB contained only small amounts of DNA. In conclusion, synthetic sites of both types contain similar but not the same sets of proteins. RB, however, have more developed microarchitecture, apparently with specific functional zones. This data suggest possible differences in genome regions replicated by these two types of replication factories. PMID- 16247618 TI - [On the relationship between psychiatry and criminal law from a legal standpoint]. AB - Psychiatry and penology are responsible together for adequate therapy of mentally ill and disordered criminals. The increased security demands of current criminal policy on the penal and forensic hospital treatment have to be moderately counteracted. The considerable increase in deculpation according to Section 21 of the German Penal Code (StGB) during the past 30 years should be balanced by more limited application, especially regarding antisocial personality disorder, borderline personality disorder, and sexual identity disorder (paraphilia). PMID- 16247619 TI - Difficulties in differentiation of Parry-Romberg syndrome, unilateral facial sclerodermia, and Rasmussen syndrome. AB - INTRODUCTION: Parry-Romberg syndrome (progressive facial hemiatrophy) is a unilateral, slowly progressive atrophy affecting the skin, subcutaneous tissues, muscles, and bones. The relationship between Parry-Romberg syndrome and connective tissue disorders, especially scleroderma en coup de sabre, is still unclear. The neurological symptoms, including epilepsy, migraine, and brain lesion, on neuroimaging may be similar. Rasmussen encephalitis (RE) is connected with chronic inflammation and damage of one hemisphere. Clinically, it is manifested by epileptic partial seizures and unilateral neurological symptoms. CASE REPORT: The authors present the case of a 10-year-old girl with features suggestive of RE, with refractory partial motor dextrolateral seizures followed by development of hemiparesis and with progressive intellectual deterioration. At the age of 2 years, some changes on the left part of the face typical of Parry Romberg syndrome or a linear form of scleroderma were noticed. DISCUSSION: The authors discussed the difficulties in differential diagnosis in that patient. The presented girl constitute the case from the borderline zone of the aforementioned disorders. PMID- 16247620 TI - Species-specific allometric scaling under self-thinning: evidence from long-term plots in forest stands. AB - Experimental plots covering a 120 years' observation period in unthinned, even aged pure stands of common beech (Fagus sylvatica), Norway spruce (Picea abies), Scots pine (Pinus sylvestris), and common oak (Quercus Petraea) are used to scrutinize Reineke's (1933) empirically derived stand density rule [see text], N=tree number per unit area, [see text]=mean stem diameter), Yoda's (1963) self thinning law based on Euclidian geometry ([see text] [see text]=mean biomass per tree), and basic assumptions of West, Brown and Enquist's (1997, 1999) fractal scaling rules ([see text] [see text] w=biomass per tree, d=stem diameter). RMA and OLS regression provides observed allometric exponents, which are tested against the exponents, expected by the considered rules. Hope for a consistent scaling law fades away, as observed exponents significantly correspond with the considered rules only in a minority of cases: (1) exponent r of [see text] varies around Reineke's constant -1.605, but is significantly different from r=-2, supposed by Euclidian or fractal scaling, (2) Exponent c of the self-thinning line [see text] roams roughly about the Euclidian scaling constant -3/2, (3) Exponent a of [see text] tends to follow fractal scaling 8/3. The unique dataset's evaluation displays that (4) scaling exponents and their oscillation are species-specific, (5) Euclidian scaling of one relation and fractal scaling of another are coupled, depending on species. Ecological implications of the results in respect to self-tolerance (common oak>Norway spruce>Scots pine>common beech) and efficiency of space occupation (common beech>Scots pine>Norway spruce>common oak) are stressed and severe consequences for assessing, regulating and scheduling stand density are discussed. PMID- 16247621 TI - Preeclampsia: increased expression of soluble ADAM 12. AB - Preeclampsia is a multisystemic pregnancy-associated disease affecting about 3-7% of pregnancies worldwide and is still a principal cause of fetal and maternal morbidity and mortality. To identify potential markers, we have compared gene expression profiles from control and preeclamptic placental tissues taken at various age-matched gestational stages using complementary DNA microarray analysis. Besides previously identified preeclampsia-associated genes, novel differentially expressed transcripts were found. The soluble form of the disintegrin metalloprotease ADAM 12 (a disintegrin and metalloproteinase 12; meltrin-alpha) represented the most upregulated transcript. This was confirmed by in situ hybridization of sections of preeclamptic placentas and by serum protein analysis of preeclamptic pregnant women. Thus, ADAM 12 could serve as an early biomarker for preeclampsia that may be of predictive and/or functional significance. PMID- 16247622 TI - Electric interactions through chirping behavior in the weakly electric fish, Apteronotus leptorhynchus. AB - The weakly electric fish Apteronotus leptorhynchus produces wave-like electric organ discharges distinguished by a high degree of regularity. Transient amplitude and frequency modulations ("chirps") can be evoked in males by stimulation with the electric field of a conspecific. During these interactions, the males examined in this study produced six types of chirps, including two novel ones. Stimulation of a test fish with a conspecific at various distances showed that two electrically interacting fish must be within 10 cm of each other to evoke chirping behavior in the neighboring fish. The chirp rate of all but one chirp type elicited by the neighboring fish was found to be negatively correlated with the absolute value of the frequency difference between the two interacting fish, but independent of the sign of this difference. Correlation analysis of the instantaneous rates of chirp occurrence revealed two modes of interactions characterized by reciprocal stimulation and reciprocal inhibition. Further analysis of the temporal relationship between the chirps generated by the two fish during electric interactions showed that the chirps generated by one individual follow the chirps of the other with a short latency of approximately 500-1,000 ms. We hypothesize that this "echo response" serves a communicatory function. PMID- 16247623 TI - Clinical evaluation of hormonal stress state in medical ICU patients: a prospective blinded observational study. AB - OBJECTIVE: To evaluate whether classification of patients as having low, moderate, or high stress based on clinical parameters is associated with plasma levels of stress hormone. DESIGN AND SETTING: Prospective, blinded, observational study in an 18-bed medical ICU. PATIENTS: Eighty-eight consecutive patients. INTERVENTIONS: Patients were classified as low (n=28), moderate (n=33) or high stress (n=27) on days 0 and 3 of ICU stay, based on 1 point for each abnormal parameter: body temperature, heart rate, systemic arterial pressure, respiratory rate, physical agitation, presence of infection and catecholamine administration. The stress categories were: high: 4 points or more, moderate 2-3 points, low 1 point. Plasma growth hormone (GH), insulin-like growth factor 1 (IGF-1), insulin, glucagon, cortisol were measured on days 0 and 3. MEASUREMENTS AND RESULTS: Plasma cortisol and glucagon were significantly higher and IGF-1 lower in high vs. low stress patients on days 0 and 3. High stress patients were more likely to have high cortisol levels (odds ratio 5.8, confidence interval 1.8-18.9), high glucagon (8.7, 2.1-36.1), and low IGF-1 levels (5.9, 1.8-19.0) than low stress patients on day 0. Moderate stress patients were also more likely to have high cortisol and glucagon levels than low stress patients. Insulin and GH did not differ significantly. Results were similar for day 3. CONCLUSIONS: Moderate and severe stress was significantly associated with high catabolic (cortisol, glucagon) and low anabolic (IGF-1) hormone levels. The hormonal stress level in ICU patients can be estimated from simple clinical parameters during routine clinical evaluation. PMID- 16247624 TI - Differential cardiovascular responses during weaning failure: effects on tissue oxygenation and lactate. AB - OBJECTIVE: To test the following two hypotheses during weaning failure: (a) mixed venous oxygen saturation (SvO2) does not decrease in patients whose oxygen consumption does not increase, and (b) blood lactate may increase in patients who demonstrate substantial decreases in SvO2. DESIGN AND SETTING: A prospective observational and physiological study in a 30-bed university intensive care unit. PATIENTS AND PARTICIPANTS: 18 patients who failed weaning and 12 patients who succeeded weaning (controls). MEASUREMENTS AND RESULTS: Hemodynamics, global tissue oxygenation, cardiovascular response (cardiac index/oxygen extraction diagram), and blood lactate were measured in ventilator-supported patients undergoing a spontaneous breathing trial. In patients who failed without having increased their oxygen consumption (n=9) the increase in oxygen delivery was accompanied by a decrease in oxygen extraction (by 15+/-4%). In patients who failed (n=9) having increased their oxygen consumption (by>10%) this increase was met mainly by an increase in oxygen extraction (by 30+/-7%). SvO2 increased by 2+/-1% in the former patients, whereas it decreased by 20+/-5% in the latter. Arterial lactate increased (range 2.3-3.1 mM/l) in only three patients who failed to have increased oxygen consumption and exhibited heart failure and the highest decreases in SvO2 (by 12-39%). CONCLUSIONS: Patients whose SvO2 does not decrease during weaning failure do not have increased oxygen consumption probably due to respiratory center depression in some of them. Patients whose SvO2 decreases have increased oxygen consumption. PMID- 16247625 TI - Weaning failure from cardiovascular origin. PMID- 16247626 TI - A new approach to DNA bending by polyamines and its implication in DNA condensation. AB - Polyamines are known to induce dynamical bending of DNA molecules. This mechanism is very important since many DNA binding proteins (DNAse, transcription factor, etc.) exert their action by their ability to bend DNA. We propose an analytical model which describes the dynamical bending of DNA by polyamine ions in highly diluted DNA solutions. The bending probability depends on the entropy loss of polyamines due to their localization. This localization is facilitated by the electrostatic repulsion between multivalent counterions condensed on DNA, which reduces the entropy loss in counterion localization. Therefore DNA bending by polyamines depends on the competition between monovalent counterions and polyamines. We find that the bending probability is weak for a low binding ratio of polyamines (i.e. number of bound polyamines per base pair), whereas a high bending probability can be reached at large polyamine binding ratio. In addition, we describe a new mechanism of DNA bending. It occurs with the help of thermal agitation, which initiates the bending and favours the polyamine localization. This model provides further insights into DNA bending by polyamines and its implication in DNA condensation. A qualitative estimation of the DNA bending probability is obtained by measuring the cleavage efficiency of DNA by bleomycin versus spermidine concentration. Indeed, a local helix distortion by polyamines results in an amplification of the double-strand cleavage by bleomycin. The measurement of the bleomycin amplification is performed by analysing images of DNA molecules with atomic force microscope. Some features of the dynamical bending indicate that condensation and bending are interrelated. PMID- 16247629 TI - SSE Spine Tango--content, workflow, set-up. www.eurospine.org-Spine Tango. AB - The Spine Tango registry is now accessible via the SSE webpage under www.eurospine.org-Spine Tango. Links to the Swiss/International, German and Austrian modules are provided as well as information about the philosophy, methodology and content. Following the links, the users are taken to the respective national modules for registration or log-in and data entry. The Swiss/International module, also accessible under www.spinetango.com, is used by all Swiss and international users, who do not have a separate national module. The physician administered forms for surgery, staged surgery and follow-up can be downloaded as PDFs.The officially recommended Spine Tango patient forms are also available. All forms were implemented in an online version and as scannable optical mark reader forms which can be ordered from the corresponding author. PMID- 16247630 TI - An unusual natural history of a L5-S1 spondylolisthesis presenting with a sacral insufficiency fracture. AB - Sacral insufficiency fractures have been described in association with conditions leading to osteoporosis. No association with spondylolisthesis has been described to date. A 60-year-old patient with known lumbosacral isthmic spondylolisthesis presented with exacerbation of symptoms initially thought to be linked to her known spinal pathology. Plain radiography, computer tomography, MRI and bone scan confirmed the presence of a recent sacral insufficiency fracture with anterior angulation. Conservative treatment resulted in improvement of symptoms after 6 months. Care should be taken when considering older patients for more aggressive treatment if they present with exacerbation of back pain and sciatica in the presence of a pre-existing spondylolisthesis. A suspicion of insufficiency fracture should be raised if risk factors exist and further investigations ordered in particular if plain radiography is normal. Lumbosacral fusion might be inappropriate in this setting. PMID- 16247631 TI - Controversies in failed back surgery syndrome. PMID- 16247632 TI - [DNA-chips in the diagnosis of hematological malignancies]. AB - In hematological malignancies, gene expression profiling using DNA-microarrays led to the discovery of novel lymphoma and leukemia subgroups. The heterogeneous entity of diffuse large B-cell lymphoma could be subdivided into the germinal center B-cell-like and the activated B-cell-like subtype which differ in pathogenesis and clinical behavior. In leukemia, existing entities defined by morphological, cytogenetic, molecular and immunophenotypic criteria were confirmed on the global gene expression level; in addition, new important molecular subgroups could be identified. In retrospective clinical lymphoma and leukemia studies, robust gene expression signatures were discovered that predict the clinical course at the time of diagnosis. Given the huge potential of the DNA microarray technology, application in the routine diagnostic setting appears possible. PMID- 16247633 TI - [Therapeutic options for nonalcoholic fatty liver disease and steatohepatitis]. AB - Nonalcoholic fatty liver disease (NAFLD) is increasingly recognized as one of the most common causes of chronic liver damage in the western world. It is strongly associated with insulin resistance, obesity and other features of the metabolic syndrome. The entity NAFLD embraces a clinical spectrum from benign steatosis over steatohepatitis to hepatic cirrhosis with its complications liver failure and hepatocellular carcinoma. Treatment is currently based on prescriptive diet and physical exercise. A well-defined pharmacotherapy of NAFLD still remains to be established due to the lack of randomized, controlled trials. Yet, for several drugs such as Metformin and Thiazolidinediones, smaller trials report promising results. PMID- 16247634 TI - Post-mortem redistribution of three beta-blockers in the rabbit. AB - To consider the role of the physico-chemical properties of drugs in their post mortem redistribution, we designed the present study to investigate the influence of lipophilicity using an experimental rabbit model. Three beta-blockers (BB), atenolol, metoprolol and propranolol, with a similar dissociation constant (pK (a)) and increasing partition coefficient (K (p)) were administered intravenously to 18 rabbits. One hour after the last administration, the animals were killed by thiopental injection and placed in a supine position at room temperature. Autopsies were performed at 0, 2, 6, 12, 24 and 48 h post-mortem. Concentrations of the three BB were determined in fluids (right and left cardiac blood, peripheral blood, urine, bile, stomach content, vitreous humour) and tissues (cardiac muscle, lungs, liver, brain, diaphragm, iliopsoas muscle) using a previously published, validated liquid chromatography-electrospray-mass spectrometry method. Our results show that lipophilicity influences post-mortem redistribution of the molecules in a certain number of anatomical sites such as the stomach, lungs, cardiac muscle, cardiac blood or liver, but does not appear to intervene in other sites such as the brain or the vitreous humour. PMID- 16247635 TI - [Innovative concepts in early cancer detection and staging of localized prostate cancer]. AB - Prostate cancer is the most common malignancy in males. Men aged 50 years and older are recommended to undergo an annual digital rectal examination (DRE) and determination of prostate-specific antigen (PSA) in serum for early detection. Fortunately, disease-specific mortality continues to decline as a result of advances in screening, staging, and patient awareness. However, about 30% of men with a clinically organ-confined disease show evidence of extracapsular extension or seminal vesicle invasion on pathological analysis. Consequently, there is a need for more accurate diagnostic tools for planning tailored treatment. A variety of modern imaging techniques has been implemented in an attempt to obtain more precise staging, thereby allowing for more detailed counseling, and instituting optimum therapy. This review highlights developments in prostate cancer imaging that may improve staging and treatment planning for prostate cancer patients. PMID- 16247637 TI - [On informed patient consent]. AB - One third of all lawsuits against doctors include statements of insufficient or lacking informed consent. The objectives of this prospective study in 104 patients were to elucidate the actual clinical routine of obtaining informed consent (process quality), collect information on active and passive recall 3 to 7 days p.op., and to investigate whether patient age, sex, education, profession, and cognitive function using the Mini Mental State Test, the time from obtaining consent to interview, acuity (emergency vs elective cases), and quantity of patient/doctor interaction would influence the patient's recall capabilities. In clinical routine, obtaining informed consent is a very variable procedure, and between two and 18 items were documented by the physician. Of the patients, 12.6% recalled actively and 43.5% passively. They named between 1.1 and 3.7 items on average, with "infection" as the leading complication, followed by "pain" and "lesion of nerves". Of all parameters investigated, only the number of initially documented items exhibit a significant effect on the patients' recall. The quantity of patient/physician interaction not only guarantees an increased effect on recall but also means improves patient interaction, thereby reducing the probability of imminent accusations. PMID- 16247636 TI - [Preoperative evaluation of prognostic factors in esophageal squamous cell cancer]. AB - Despite substantial improvements in the surgical therapy of esophageal squamous cell cancer, the prognosis still remains poor. This is mainly due to locally advanced tumors (T3-4, N+) or systemic metastases (M1) in the majority of patients at initial presentation. It is of the utmost importance to reliably detect relevant pretherapeutic prognostic indicators for optimal individual therapeutic strategies. Pretherapeutic prognostic indicators should therefore discriminate precisely between incurable and potentially curative disease. Preoperative or definitive multimodal treatment is increasingly being offered to patients with locally advanced tumors and opens a broad field for innovative techniques such as pretherapeutic molecular response prediction or early response detection by PET scan to further individualize and optimize treatment strategies. PMID- 16247638 TI - [Acute aortic dissection. Differential diagnosis of a thoracic emergency]. AB - Acute aortic dissection is an infrequent but important differential diagnosis of acute chest pain. The variability of presenting symptoms makes it difficult to diagnose correctly. Important clinical indicators - besides chest pain - are symptoms related to acute aortic insufficiency and/or pericardial tamponade, variable acute neurologic alterations, or signs of peripheral or visceral malperfusion. The spontaneous prognosis depends on the location and extent of the dissection, and left untreated dissection carries a high mortality. The key goal of preclinical treatment is stabilization with analgesia, mild sedation (opioids, benzodiazepines) and treatment of hypertension (beta-blockers) or hypotension (fluid administration). If the patient presents with a high probability of dissection, early transfer to a specialized center appears advisable. Initial clinical diagnostic studies include transthoracic echocardiogram and computed tomography. If the ascending aorta is involved (Stanford type A) immediate replacement of the proximal aorta is necessary. Isolated dissections of the descending aorta (type B) require aggressive blood pressure control, but can be managed conservatively in most cases. A high level of vigilance is necessary in all patients to detect and treat visceral ischemia. PMID- 16247639 TI - [Expert evidence. General prerequisites and procedural characteristics for expert opinions]. AB - In view of the boom in medical liability suits, which centre around medical malpractice, the decision of the medical expert is the pivot in the conflict between doctor and patient. Therefore, from a legal point of view it is extremely important that the expert is well prepared for a courtroom appearance. Beginning from a systematic and logical structure of the expert opinion, the most important demands on quality are dealt with: professional competence, objectivity and impartiality, intellectual integrity and intelligibility of speech, duty to give a strictly personal delivery of the opinion, limitation to the medical specialty, no legal comments, no insinuation and strict adherence to the theme. As an aid to the Judge, the expert must have a certain degree of background legal knowledge. This includes the peculiarities of civil and criminal procedures with respect to the burden of proof, the causality of medical or organisational errors in treatment leading to injury or even death of the patient, the significance of the term "a probability bordering on certainty", the meaning of "gross" errors in treatment, the guidelines and the medical standards. The obligation of the medical expert in questions of medical malpractice is not only an extremely responsible and difficult one, but also not without risk for the expert himself. Since August 2002 gross negligent errors by the expert are also liable for compensation in law under [section sign] 839a of the German Civil Code Book. PMID- 16247640 TI - POEMS syndrome: radiographic appearance with MRI correlation. AB - POEMS syndrome is a rare disorder in which patients present with the hallmark signs of polyneuropathy, organomegaly, endocrinopathy, M protein and skin changes. Many other clinical findings are also often present, most notably osseous lesions. The MRI appearance of the bony lesions in POEMS syndrome has been described in five cases, four of which are in the non-English literature. We report the MRI appearance of the osseous lesions in a patient with POEMS syndrome who presented with sciatic neuropathy. PMID- 16247641 TI - Accuracy of radiography in grading and tissue-specific diagnosis--a study of 200 consecutive bone tumors of the hand. AB - OBJECTIVE: To determine the usefulness of radiography and magnetic resonance imaging in differentiating benign from malignant bony tumors of the hand and in making a tissue-specific diagnosis. DESIGN: Two hundred consecutive bony tumors of the hand, the details of which originated from a national databank, were studied in a prospective way by radiography (100%) and by MRI (25%). All tumors were graded on a five-point scale, from certainly benign to certainly malignant, using location and morphology as diagnostic parameters. For all tumors a tissue specific diagnosis was made, by the proposal of three possibilities in decreasing order of probability. Histological diagnosis was made by peer review, according to the WHO classification. RESULTS: By the combining of "certainly" and "probably" benign (grades I and II) and "certainly" and "probably" malignant (grades IV and V), a correct grading was obtained in 165 (82.5%) of the cases (154 of the 173 benign and 11 of the 27 malignant tumors). A correct tissue specific diagnosis was included in the three proposed differentials in 87.5%. MRI confirmed a correct diagnosis made on radiography in 72% and improved the grading capability by correctly upgrading malignant tumors and downgrading benign tumors in, respectively, 8% and 12%. The capability to obtain a tissue-specific diagnosis improved with change of an incorrect diagnosis on radiography to a correct one on MRI in 12 cases (24%). CONCLUSION: Subjective (semiquantitative) grading on radiography by an expert group proved to be excellent when compared with the results of a quantitative analysis of individual grading parameters. Multiple logistic regression analysis of these parameters resulted in a grading formula containing only six variables. The additional value of MRI in grading was amply demonstrated. Already high accuracy of radiography, in making a tissue specific diagnosis, improved substantially after the performance of MRI. PMID- 16247642 TI - Bone mineral measurements of subchondral and trabecular bone in healthy and osteoporotic rabbits. AB - INTRODUCTION: Experimental models of osteoporosis in rabbits are useful to investigate anabolic agents because this animal has a fast bone turnover with predominant remodelling over the modelling processes. For that purpose, it is necessary to characterize the densitometric values of each type of bony tissue. OBJECTIVE: To determine areal bone mass measurement in the spine and in trabecular, cortical and subchondral bone of the knee in healthy and osteoporotic rabbits. DESIGN: Bone mineral content and bone mineral density were measured in lumbar spine, global knee, and subchondral and cortical bone of the knee with dual energy X-ray absorptiometry using a Hologic QDR-1000/W densitometer in 29 skeletally mature female healthy New Zealand rabbits. Ten rabbits underwent triplicate scans for evaluation of the effect of repositioning. Osteoporosis was experimentally induced in 15 rabbits by bilateral ovariectomy and postoperative corticosteroid treatment for 4 weeks. Identical dual energy X-ray absorptiometry (DXA) studies were performed thereafter. RESULTS: Mean values of bone mineral content at the lumbar spine, global knee, subchondral bone and cortical tibial metaphysis were: 1934+/-217 mg, 878+/-83 mg, 149+/-14 mg and 29+/-7.0 mg, respectively. The mean values of bone mineral density at the same regions were: 298+/-24 mg/cm(2), 455+/-32 mg/cm(2), 617+/-60 mg/cm(2) and 678+/-163 mg/cm(2), respectively. Bone mineral content and bone density of healthy rabbits followed a normal distribution at the four skeletal regions studied. Precision after triplicate repositioning yielded a coefficient of variation ranging from 2.6% to 3.8%. The least significant change ranged between 7.3% and 10.7%. Bone mineral density measured at the four different skeletal regions correlated significantly. Bone mineral density in osteoporotic rabbits was significantly lower in the four regions studied than that in controls, rendering a T-score of, respectively, 2.0+/-1.1 in the lumbar spine, -2.2+/-2.1 in the global knee, -1.9+/-0.6 in the subchondral bone, and -5.7+/-3.1 in the cortical tibia (P<0.05). CONCLUSIONS: DXA is a reliable and precise method to evaluate the bone mass in rabbits. Our results also suggest that subchondral bone is a bone of mixed densitometric characteristics with marked cortical bone predominance. PMID- 16247643 TI - Phosphodiesterase inhibitors in female sexual dysfunction. AB - Based on the increasing knowledge on both the physiology of penile erection and the pathophysiology of erectile dysfunction, selective phosphodiesterase (PDE) inhibitors have been successfully introduced in the oral treatment of male erectile dysfunction. Because of their central role in smooth muscle tone regulation, PDEs remain an attractive target for drug development in urology. Since the distribution and functional significance of PDE isoenzymes vary in different tissues, selective inhibitors of the isoenzymes have the potential to exert at least partially specific effects on the target tissue. Currently, PDE inhibitors are under investigation with potential uses in urinary stone disease, overactive bladder and the so-called benign prostatic syndrome. The convincing clinical data on the use of the orally active PDE5 inhibitors sildenafil (VIAGRA), vardenafil (LEVITRA) and tadalafil (CIALIS) in the treatment of erectile dysfunction are accompanied by boosting research activities on intracellular signal transduction and PDE characterisation in female genital tissues with the aid of immunohistochemistry and immunocytochemistry and molecular biology. The expression of various PDE isoforms in the human clitoris, vagina and labia minora was shown by means of immunohistochemistry and RT-PCR analyses and it was concluded from functional studies that an increase in cGMP or cAMP might be involved in the regulation of female genital blood flow and the control of genital non-vascular smooth muscle. As a consequence, the efficacy and safety of the PDE5 inhibitor sildenafil in the treatment of symptoms of female sexual dysfunction (FSD), including female sexual arousal disorders (FSAD), have been evaluated. Although the experiences from these early clinical studies have so far not been conclusive, they suggest that, after appropriate evaluation of patients, inhibition of PDE5 might be of benefit for selected individuals with FSAD. Such research efforts will possibly allow the identification of efficacious and diagnostic tools for erectile dysfunction and of even more selective drugs in its therapy. PMID- 16247645 TI - Long-term follow-up of atypical membranoproliferative glomerulonephritis: are steroids indicated? AB - Atypical membranoproliferative glomerulonephritis (MPGN) has been reported to have a good prognosis when treated with corticosteroids. However, this recommendation is based on uncontrolled trials and is associated with many complications. The purpose of our study is to determine whether steroid therapy is indicated for atypical MPGN. The cases of seven patients with atypical MPGN are reported in this study. Urinary abnormalities of five of them were detected by urine screening at school, of two because of macrohematuria. Hypocomplementemia was noted in six patients. All but one patient were treated without corticosteroids, and five with angiotensin-converting enzyme inhibitors (ACEI) and/or the Chinese herbal medicine Sairei-to (TJ-114). One patient recovered spontaneously from proteinuria and was therefore not treated, and one who developed severe proteinuria during observation was treated with corticosteroids. After an average follow-up period of 10.0 years, five patients showed normal urinary findings, one had hematuria and one proteinuria. At the most recent follow-up, the renal function of all patients remained within the normal range, and serum C3 had returned to normal levels in five out of six. These findings suggest that the indication of steroid therapy for atypical MPGN should be re-examined, since most of the patients with atypical MPGN seem to have an excellent prognosis without treatment with corticosteroids. PMID- 16247644 TI - Prevention and treatment of renal osteodystrophy in children on chronic renal failure: European guidelines. AB - Childhood renal osteodystrophy (ROD) is the consequence of disturbances of the calcium-regulating hormones vitamin D and parathyroid hormone (PTH) as well as of the somatotroph hormone axis associated with local modulation of bone and growth cartilage function. The resulting growth retardation and the potentially rapid onset of ROD in children are different from ROD in adults. The biochemical changes of ROD as well as its prevention and treatment affect calcium and phosphorus homeostasis and are directly associated with the development of cardiovascular disease in pediatric renal patients. The aims of the clinical and biochemical surveillance of pediatric patients with CRF or on dialysis are prevention of hyperphosphatemia, avoidance of hypercalcemia and keeping the calcium phosphorus product below 5 mmol(2)/l(2). The PTH levels should be within the normal range in chronic renal failure (CRF) and up to 2-3 times the upper limit of normal levels in dialysed children. Prevention of ROD is expected to result in improved growth and less vascular calcification. PMID- 16247647 TI - Intussusception of the small bowel associated with nephrotic syndrome. AB - We report the case of a 2-year-old boy who developed a small bowel intussusception during treatment failure of his first episode of nephrotic syndrome. Despite the absence of typical symptoms other than abdominal pain, the intussusception was diagnosed by ultrasonography and computed tomography and successfully reduced by air enema. No pathological lead point was discovered, and no symptoms of Henoch-Schonlein purpura developed later. Intussusception should be considered in the differential diagnosis of abdominal pain in patients with nephrotic syndrome, especially in patients exhibiting prolonged edema. Ultrasonography or computed tomography should be performed, even in the absence of other typical symptoms suggestive of intussusception. We should also bear in mind that the intussusception associated with nephrotic syndrome might occur at regions other than the typical ileocolic region, such as within the small intestine. PMID- 16247646 TI - Renal transplantation in a 14-year-old girl with vitamin B12-responsive cblA-type methylmalonic acidaemia. AB - Renal tubular dysfunction and chronic renal failure are well recognised complications of methylmalonic acidaemia (MMA) and can occur even in the context of optimal medical metabolic management. Organ transplantation, such as renal and combined liver and renal transplants, have been utilised in the past for children whose disease cannot be managed by conservative medical practices and those with end stage renal disease. Our patient was diagnosed with B(12)-responsive MMA (subsequently proven to be cblA-type MMA) in the postoperative period following renal transplantation for idiopathic chronic renal failure. She remains well, with excellent graft function and metabolic control 4 years after transplantation. This patient highlights the importance of testing for the inborn errors of metabolism in patients presenting with recurrent acidosis and progressive renal impairment. PMID- 16247648 TI - C1Q nephropathy in children. AB - C1q nephropathy (C1qNP) is a peculiar form of glomerulonephritis characterized by mesangial immunoglobulin and complement deposits, predominantly C1q, with no evidence of systemic lupus erythematosus. We describe the incidence, manifestation, histopathologic findings, follow-up, treatment and outcome of C1qNP. Twelve C1qNP patients were identified among 131 children who had undergone renal biopsy, accounting for a 9.16% incidence of C1qNP. Light microscopy examination showed focal segmental glomerulosclerosis (FSGS) with or without diffuse mesangial proliferation (n=6), minimal change disease (MCD) (n=4) or focal glomerulonephritis (n=2). C1q deposits were found in all, while electron microscopy revealed visible deposits in nine cases. Eight children presented with nephrotic syndrome, while one had nephrotic proteinuria and renal insufficiency that progressed to end-stage renal failure. The remaining three patients presented with nonnephrotic proteinuria associated with microhematuria, hypertension or renal insufficiency. Only one nephrotic syndrome patient responded excellently to corticosteroids, while four became corticosteroid dependent, and three were corticosteroid resistant, showing a very poor response to other immunosuppressive therapy as well. Patients with non-nephrotic proteinuria demonstrated fixed laboratory findings. Most C1qNP patients had FSGS or MCD, the majority of them presenting with corticosteroid-dependent or corticosteroid-resistant nephrotic syndrome. The latter showed a very poor response to any immunosuppressive therapy and high risk for progressive renal insufficiency. PMID- 16247650 TI - Review of the literature regarding the relationship of rebleeding and external ventricular drainage in patients with subarachnoid hemorrhage of aneurysmal origin. AB - Acute hydrocephalus is a well-documented complication of subarachnoid hemorrhage. The insertion of external ventricular drainage (EVD) has been the standard of care in the management of this complication, aiming primarily at immediate improvement of the clinical condition of these patients, making them more suitable candidates for surgical or endovascular intervention. In our current communication, we review the pertinent literature regarding the relationship of rebleeding and EVD. Several studies have implicated a significantly increased risk of rebleeding in patients with EVD, compared with patients without it. Abrupt lowering of the intracranial pressure could lead to rebleeding due to decreased transmural pressure or removal of the clot sealing the previously ruptured aneurysm. However, a variety of parameters that could affect the rebleeding rate, such as the timing of surgery, the timing and duration of drainage, the size of the aneurysm, as well as the severity of the initial hemorrhage, do not seem to have been adequately explored in the majority of these studies. In addition, a number of clinical trials have failed to provide evidence for the negative role of EVD in the development of rebleeding. Conclusively, further long-term multi-center studies are required in order to establish the exact nature of the relationship between EVD and rebleeding after aneurysmal subarachnoid hemorrhage. PMID- 16247652 TI - Amaranthin in feather cockscombs is synthesized via glucuronylation at the cyclo DOPA glucoside step in the betacyanin biosynthetic pathway. AB - Uridine 5'-diphosphate (UDP)-glucuronic acid: cyclo-DOPA 5-glucoside glucuronosyltransferase activity was detected in a crude extract prepared from the purple flowers of feather cockscombs. This suggests that the glucuronic acid moiety of amaranthin and its derivatives may be introduced at the cyclo-DOPA glucoside step, but not at the betanidin glucoside step. PMID- 16247653 TI - Low genetic diversity and significant population structuring in the relict Amentotaxus argotaenia complex (Taxaceae) based on ISSR fingerprinting. AB - Amentotaxus, a genus of the Taxaceae, represents an ancient lineage that has long existed in Eurasia. All Amentotaxus species experienced frequent population expansion and contraction over periodical glaciations in Tertiary and Quaternary. Among them, Amentotaxus argotaenia complex consists of three morphologically alike species, A. argotaenia, Amentotaxus yunnanensis, and Amentotaxus formosana. This complex is distributed in the subtropical region of mainland China and Taiwan where many Pleistocene refugia have been documented. In this study, genetic diversity and population structuring within and between species were investigated based on the inter-simple sequence repeats (ISSR) fingerprinting. Mean genetic diversity within populations was estimated in three ways: (1) the percentage of polymorphic loci out of all loci (P) (2) Nei's unbiased expected heterozygosity (He), and (3) Shannon's index of phenotypic diversity. For a total of 310 individuals of 15 populations sampled from the three species, low levels of ISSR genetic variation within populations were detected, with P=4.66-16.58%, He=0.0176-0.0645 and Hpop=0.0263-0.0939, agreeing with their seriously threatened status. AMOVA analyses revealed that the differences between species only accounted for 27.38% of the total variation, whereas differences among populations and within populations were 57.70 and 14.92%, respectively, indicating substantial isolation between the patch-like populations. A neighbor joining tree identified a close affinity between A. yunnanensis and A. formosana. Genetic drift due to small population size, plus limited current gene flow, resulted in significant genetic structuring. Low levels of intra-populational genetic variation and considerable inter-populational divergence were also attributable to demographic bottlenecks during and/or after the Pleistocene glaciations. PMID- 16247651 TI - Neuropsychopharmacological properties of neuroactive steroids in depression and anxiety disorders. AB - Neuroactive steroids modulate neurotransmission through modulation of specific neurotransmitter receptors such as gamma-aminobutyric acid type A (GABAA) receptors. Preclinical studies suggested that neuroactive steroids may modulate anxiety- and depression-related behaviour and may contribute to the therapeutical effects of antidepressant drugs. Attenuations of 3alpha-reduced neuroactive steroids have been observed during major depression. This disequilibrium can be corrected by successful treatment with antidepressant drugs. However, non pharmacological antidepressant treatment strategies did not affect neuroactive steroid composition independently from the clinical response. Further research is needed to clarify whether enhancement of neuroactive steroid levels might represent a new therapeutical approach in the treatment of affective disorders. Nevertheless, the first studies investigating the therapeutical effects of exogenously administered dehydroepiandosterone revealed promising results in the treatment of major depression. In addition, in various anxiety disorders alterations of neuroactive steroid levels have been observed. In panic disorder, in the absence of panic attacks, neuroactive steroid composition is opposite to that seen in depression, which may represent counter-regulatory mechanisms against the occurrence of spontaneous panic attacks. However, during experimentally induced panic attacks, there was a pronounced decline in GABAergic neuroactive steroids, which might contribute to the pathophysiology of panic attacks. In conclusion, neuroactive steroids contribute to the pathophysiology of affective disorders and the mechanisms of action of antidepressants. They are important endogenous modulators of depression and anxiety and may provide a basis for the development of novel therapeutic agents in the treatment of affective disorders. PMID- 16247654 TI - Choice of flaps for breast reconstruction. AB - The optimal method for breast reconstruction should be safe, reliable, and accessible for every patient, and it should display little or no donor-site morbidity. After comparing mammary implants it has been found that autogenous breast reconstruction can create a ptotic, soft, symmetrical breast mound. The transverse rectus abdominis musculocutaneous flap (TRAM) remains the most popular method for autogenous reconstruction. Modern trends in breast reconstruction using the TRAM flap have promoted adequate blood supply to the flap while minimizing donor-site defects in the anterior abdominal wall. The pedicled TRAM flap remains one of the most frequently used flaps, but the indirect blood supply in this flap has required many modifications and refinements. Such modifications have included the bipedicled TRAM flap, the free TRAM flap, and the supercharged TRAM flap. To avoid donor-site morbidities, the muscle-sparing free TRAM, deep inferior epigastric perforator flap (DIEP), and superficial inferior epigastric artery (SIEA) flap were introduced. The DIEP perforator flap requires meticulous technique but offers proven reliability and a low rate of complications. As surgeons become more comfortable with harvesting DIEP flaps, the frequency of usage seems likely to increase. The latissimus dorsi musculocutaneous flap, gluteus maximus musculocutaneous flap, and others may be selected when these modifications of free TRAM flap are unavailable or unusable. PMID- 16247655 TI - Cosmetic reconstruction after resection of breast cancer: use of the ELD-MC flap and EVRAM flap. AB - Breast reconstruction following the resection of breast cancer with inadequate residual chest-wall tissue may be performed with an implant or a myocutaneous flap, such as the latissimus dorsi or a rectus abdominis. Among a variety of operative procedures, each method has advantages and disadvantages. The insertion of a silicone-bag prosthesis is the easiest method, but the prosthetic implant sometimes has complications, such as unfavorable capsular contracture formation around the implant, rupture, infection, or exposure. We therefore use an extended latissimus dorsi myocutaneous (ELD-MC) flap with some amount of surrounding subcutaneous fat from the lumbar area, and avoid the use of any implant with an MC flap. Also, for the reconstruction and correction of infraclavicular and axillary depression, we use the extended vertical rectus abdominis myocutaneous (EVRAM) flap. This method uses the skin and fat on both sides of the umbilicus as a lenticular flap vascularized by only one of the rectus abdominis muscles. The patients are satisfied with the outcome because symmetry and good breast volume can be obtained. There have been no functional or anatomical defects of the donor area. No abdominal hernia after an EVRAM flap has resulted to date. Both the ELD MC and EVRAM flaps can be successfully used for cosmetic breast reconstruction after the resection of breast cancer. PMID- 16247656 TI - Treatment of lymphedema with lymphaticovenular anastomoses. AB - Although lymphedema in the extremities is a troublesome adverse effect following radical resection of various cancers, conventional therapies for lymphedema are not always satisfactory, and new breakthroughs are anticipated. With the introduction of supermicrosurgical techniques for the anastomosis of blood or lymphatic vessels less than 0.8 mm in diameter, we have developed a novel method of lymphaticovenular anastomosis for the treatment of primary as well as secondary lymphedema in the extremities. Here, we review the pathophysiological aspects of lymphedema, emphasizing the importance of smooth-muscle cell function in the affected lymphatic walls. We then describe the theoretical basis and detailed operative techniques of our lymphaticovenular anastomoses. Although technically demanding, especially for beginners, we believe that this method will become a new clinical standard for the treatment of lymphedema in the near future. PMID- 16247657 TI - Treatment strategy for cutaneous malignant melanoma. AB - The incidence of cutaneous malignant melanoma has been rising in Japan. With education, recent advances in accurate diagnosis and establishment of the concept, more lesions are being diagnosed as early melanomas, for which there is a high cure rate. However, many patients will still present with thicker lesions or nodal involvement, which carries a significantly worse prognosis. Recently there have been advances in the management and treatment of cutaneous melanoma. This article reviews the clinical evidence behind the current treatment recommendations for primary and recurrent cutaneous melanoma in Japan. PMID- 16247658 TI - Role of VEGF-C and VEGF-D in lymphangiogenesis in gastric cancer. AB - BACKGROUND: The molecular mechanisms of lymphangiogenesis induced by vascular endothelial growth factor (VEGF)-C and VEGF-D in gastric cancer were studied. METHODS: VEGF-C and VEGF-D gene expression vectors were transfected into the gastric cancer cell line KKLS, which did not originally express VEGF-C and VEGF D, and stable transfectants (KKLS/VEGF-C and KKLS/VEGF-D) were established. The cell lines were inoculated into the subserosal layer of the stomach and subcutaneous tissue of nude mice. RESULTS: VEGF-C and VEGF-D expression in KKLS/VEGF-C and KKLS/VEGF-D cells was found by reverse transcription polymerase chain reaction (RT-PCR) and Western blot analysis. Expression of mouse VEGF receptor (VEGFR)-2 and mouse VEGFR-3 mRNA was detected in the KKLS/VEGF-C and KKLS/VEGF-D gastric tumors. Newly formed lymphatic vessels were detected not only in the periphery but also in the center of the tumors. The intratumor lymphatic vessels connected with the preexisting lymphatic vessels in the muscularis mucosa. The average numbers of lymphatic vessels in KKLS/VEGF-C (52.0 +/- 9.5) and KKLS/VEGF-D (16.4 +/- 0.6) gastric tumors were significantly higher than that in the KKLS/control vector tumors (4.0 +/- 1.4). CONCLUSION: VEGF-C and VEGF-D may induce neoformation of lymphatic vessels in experimental gastric tumors by the induction of VEGFR-3 expression. PMID- 16247659 TI - In vitro detection of cross-resistant and non-cross-resistant agents with fluorouracil for patients with colorectal cancer. AB - BACKGROUND: Fluorouracil-based chemotherapy, such as that with 5-fluorouracil (5 FU)/leucovorin, is standard as first-line chemotherapy for advanced colorectal cancer (CRC) in Japan. However, the best agent for second-line chemotherapy after fluorouracil failure is yet to be determined. This study was undertaken to find an appropriate agent for second-line chemotherapy. METHODS: Seventy-five tumor specimens from CRC patients with no prior chemotherapy were obtained operatively and their chemosensitivity to five anticancer agents; i.e., 5-FU, mitomycin C (MMC), cisplatin, docetaxel, and an active metabolite of irinotecan (SN-38), was analyzed in an in vitro chemosensitivity test. In this method, the degree of chemosensitivity was expressed as the percent T/C ratio, where T was the total volume of the tumor colonies in the treated group and C was that of the control group. Pearson's correlation coefficients were used to assess the relationship between two agents. RESULTS: Fifty-eight specimens (colon, 28; rectum, 30) were successfully analyzed. Positive correlations with 5-FU chemosensitivity were verified for the chemosensitivity of MMC, cisplatin, and docetaxel. No correlation with 5-FU chemosensitivity was verified for SN-38 chemosensitivity. Although the functional mechanism of each of the agents differs from that of 5 FU, with the exception of irinotecan, they all had a spectrum closely similar to the 5-FU spectrum. CONCLUSION: Only irinotecan exhibited a spectrum independent of that of 5-FU, thus indicating that it could be an appropriate agent for second line chemotherapy after fluorouracil failure. PMID- 16247660 TI - Monthly paclitaxel and carboplatin with oral estramustine phosphate in patients with hormone-refractory prostate cancer. AB - BACKGROUND: We aimed to determine the safety and efficacy of monthly paclitaxel and carboplatin with oral estramustine phosphate in patients with hormone refractory prostate cancer (HRPC). METHODS: Patients with prostate cancer that was progressing despite androgen ablation therapy were treated with i.v. paclitaxel, 175 mg/m2, over 3 h, followed by carboplatin (area under the curve, 5) on day 1, with oral estramustine phosphate, 280 mg twice daily, for a 28-day treatment cycle. Estramustine phosphate was precluded in those patients who had experienced adverse effects during prior chemotherapies. Patients were evaluated for response every cycle, and the treatment was continued until the cancer progressed. RESULTS: Twenty-one patients with progressive hormone-refractory disease were treated for a median of 4 cycles (range, 1 to 11 cycles). Estramustine phosphate was precluded in seven patients. Post-therapy decreases in serum prostate-specific antigen levels of 50% and 75%, respectively, were seen in 43% and 19% of the patients (95% confidence intervals, 22% to 64% and 2% to 36%). Of the nine patients with measurable disease, 1 (11%) had a complete response and 2 (22%) had a partial response. The overall median time to progression was 4 months, and the median survival time for all patients was 11 months. Major grade 3 or 4 adverse effects were anemia (29%), neutropenia (48%), and thrombocytopenia (24%). Mild peripheral neuropathy and myalgia/arthralgia were observed in 11 (52%) and 9 (43%) patients, respectively. CONCLUSION: Monthly paclitaxel and carboplatin with oral estramustine phosphate has significant antitumor activity and is well tolerated in patients with progressive HRPC. PMID- 16247661 TI - Clinical outcome of combined immunotherapy with low-dose interleukin-2 and interferon-alpha for Japanese patients with metastatic renal cell carcinoma who had undergone radical nephrectomy: a preliminary report. AB - BACKGROUND: The objective of this study was to evaluate the clinical outcome of combined immunotherapy with interferon-alpha (IFN-alpha) and low-dose interleukin 2 (IL-2) for Japanese patients with metastatic renal cell carcinoma (RCC) who had undergone radical nephrectomy. METHODS: This study included 13 patients who were diagnosed as having metastatic RCC following radical nephrectomy. These patients received a subcutaneous injection of IFN-alpha (6 x 10(6) IU per day) three times per week and an intravenous injection of IL-2 (1.4 x 10(6) IU per day) twice per week. Tumor response was evaluated every 16 weeks, and as a rule, this weekly regimen was repeated 50 times in patients with evidence of objective response or stable disease. RESULTS: One of the 13 patients dropped out because of severe toxicity; hence, 12 patients were evaluable, with a median follow-up period of 18 months after the start of this combined therapy. Six patients (50.0%) achieved objective responses, with 1 complete response (CR), while only 2 (16.7%) demonstrated progressive disease. The median duration of response in the 6 responders was 13.5 months. Toxicity associated with this combined immunotherapy was limited to WHO grade 1 or 2 in these 12 patients. All patients were alive at last follow-up, and 2 remain disease-free after 1 additional patient showed a CR following surgical resection of the remaining metastatic disease. CONCLUSION: Our preliminary experience suggests that long-term, repeated treatment with IFN-alpha and low-dose IL-2 is feasible in Japanese patients with metastatic RCC who have undergone radical nephrectomy. Although it will be necessary to accumulate data from a larger number of patients with a longer follow-up period, the combined immunotherapy tested in this study may become the preferred therapy for Japanese patients with metastatic RCC. PMID- 16247662 TI - Pancreatic and gastric metastases of leiomyosarcoma arising in the left leg. AB - Pancreatic or gastric metastases from other primary malignancies are rare, especially from leiomyosarcoma. We report a case of leiomyosarcoma in the left lower leg with metastases to the pancreas and stomach. A 61-year-old man had liver cirrhosis caused by hepatitis C virus infection and was followed up by his primary physician. Two years before presentation at our hospital, he had undergone surgical resection of leiomyosarcoma in the left lower leg and systemic chemotherapy for multiple metastatic tumors in the lung. On admission, endoscopic examination and computed tomography were performed for a routine checkup to exclude esophageal varices and liver tumor. Although the patient had no specific symptoms, multiple gastric and pancreatic metastases were identified by endoscopy and computed tomography, respectively. In general, metastases to the pancreas and stomach are rare. We discuss the clinical and diagnostic findings of pancreatic and gastric metastases by reviewing previously reported cases. PMID- 16247664 TI - Recurrent retroperitoneal malignant nerve sheath tumor associated with neurofibromatosis type 1 responding to carboplatin and etoposide combined chemotherapy. AB - A 25-year-old man was referred to our hospital with left flank pain, and computed tomography (CT) and magnetic resonance imaging (MRI) revealed large retroperitoneal masses. Physical examination revealed many cafe-au-lait spots and superficial neurofibromas, and a diagnosis of neurofibromatosis type 1 (von Recklinghausen's disease) was made. The tumor was resected, and the pathological diagnosis was malignant peripheral nerve sheath tumor (MPNST). Six months after the operation, lung metastases were detected. Surgical resection was incomplete, as there were too many lesions. He received four courses of chemotherapy with carboplatin and etoposide, and the metastatic lung lesions were markedly decreased. After chemotherapy, complete resection of the remaining lung lesions was performed, and there has been no recurrence to date. PMID- 16247663 TI - Gastric small cell carcinoma with marked response to neoadjuvant chemotherapy. AB - We report a patient with gastric small cell carcinoma (SCC) who showed a marked response to neoadjuvant chemotherapy. The patient was a 72-year-old Japanese man who was admitted because of epigastralgia. Subsequent examination revealed the presence of advanced gastric carcinoma in the lesser curvature of the lower body of the stomach, with multiple abdominal lymph node metastases. Endoscopic biopsy specimens from the tumor revealed SCC with moderately differentiated adenocarcinoma. The patient received neoadjuvant chemotherapy consisting of carboplatin (400 mg/m2, for 1 day), epirubicin (27 mg/m2, for 1 day), etoposide (70 mg/m2, for 3 days), and 5-fluorouracil (330 mg/m2, for 11 days). Clinically, the primary tumor and lymph node metastases were markedly reduced. Subsequently the patient underwent curative surgery and was alive without recurrence for more than 3 years after the surgery. PMID- 16247665 TI - Excellent response of chemotherapy-resistant B-cell-type chronic lymphocytic leukemia with meningeal involvement to rituximab. AB - A 70-year-old woman was diagnosed with B-cell-type chronic lymphocytic leukemia (B-CLL) in May 2001. Initial white blood cell (WBC) count was 37 x 10(9)/l and most of the cells were mature small lymphocytes. Surface antigen analysis of these lymphocytes revealed positive reactions for CD19, 20, 25, 5, and lambda light chain. Despite her Rai stage-0 status, various treatments were ineffective, including cyclophosphamide; fludarabine; 6-mercaptopurine; a combination of vincristine, cyclophosphamide, prednisolone, and adriamycin; and etoposide. Her WBC count increased, ranging from 150 to 450 x 10(9)/l, with marked splenomegaly, and symptoms of meningitis, such as headache, ophthalmalgia, hearing disturbance, and abnormal behavior, being manifested. The WBC count in the cerebrospinal fluid was elevated to 134/microl. The surface phenotype of these cells was identical to that of circulating lymphocytes, indicating meningeal involvement of leukemia, a rare complication in B-CLL. At the time of this WBC elevation, 24% of circulating lymphocytes had prominent nucleoli, indicating progression of the disease to CLL/prolymphocytic leukemia. Her symptoms disappeared after repeated intrathecal injections of methotrexate and dexamethazone. After four courses of treatment of the refractory B-CLL with rituximab, an anti-CD20 monoclonal antibody, the WBC count returned to normal levels and the splenomegaly disappeared. She is currently well, with sustained remission, as of April 2004. PMID- 16247666 TI - Advanced clear-cell adenocarcinoma of the bladder successfully treated by radical surgery with adjuvant chemoradiotherapy. AB - A 52-year-old woman was referred to our institute for the evaluation of a tumor in her pelvic cavity. The tumor seemed to have arisen from the bladder or urethra, and bilateral iliac lymphadenopathy was seen. Her urethral mucosa looked intact according to the results of cystourethroscopy. Histopathological examination of the biopsy specimens showed clear-cell adenocarcinoma. She underwent radical cystourethrectomy with complete pelvic lymph node dissection and the construction of a bilateral ureterocutaneostomy. Macroscopically, the tumor had arisen from the trigone of the bladder, and histopathological examination of the tumor revealed adenocarcinoma exhibiting solid clear cells with glandular and papillary patterns. The tumor had infiltrated perivesical structure (pT3a), and metastases in multiple pelvic lymph nodes were recognized (pN3). Postoperatively, three courses of systemic combination chemotherapy with 5 fluouracil (FU) and cisplatin, along with a total of 45 Gy of irradiation during the second course of chemotherapy, were conducted. No evidence of the disease has been seen 28 months after the surgery. PMID- 16247667 TI - Coexistence of metastatic lung cancer and pulmonary tuberculosis diagnosed in the same cavity. AB - A 79-year-old woman who had a past history of chronic renal failure 10 years earlier, tongue cancer (T2N2M0) 3 years earlier, and tuberculosis of the cervical lymph nodes 6 months earlier was suddenly admitted with the complaint of right chest pain on April 6, 2004. Right pneumothorax and mild pleural effusion were observed on a chest radiograph. There was no improvement in the patients collapsed lung despite the insertion of a chest drainage tube into the pleural cavity. Three thin-walled cavitary lesions were noted in the right lobe of segment 1 on computed tomography, and the cause of her pneumothorax was thought to be air leakage from the largest cavitary lesion adjacent to the visceral pleura. Partial resection of the right lung by video-assisted thoracoscopic surgery (VATS) was performed at the Department of Thoracic Surgery. Subsequently, it was determined that metastatic squamous cell carcinoma of the lung, corresponding to her tongue cancer, had invaded the visceral pleura adjacent to the largest cavitary lesion. Simultaneously, an epitheloid granuloma with caseating necrosis was observed adjacent to a partially thickened portion of this cavitary lesion. The epitheloid granuloma was found to be acid-fast bacilli positive and a diagnosis of Mycobacterium tuberculosis pulmonary tuberculosis was made. We report a rare case of the coexistence of metastatic lung cancer originating from tongue cancer and active pulmonary tuberculosis diagnosed in the same large cavitary lesion. PMID- 16247668 TI - Nisin biosynthesis and its properties. AB - The antimicrobial peptide, nisin, produced by several strains of Lactococcus lactis, which belongs to the Class I bacteriocins called lantibiotics, is a small (3.4 kDa), 34-amino acid, cationic, hydrophobic peptide and has the five characteristic (beta-methyl)lanthionine rings formed by significant post translational modification. A cluster of 11 genes has been involved in the biosynthesis of nisin and are proposed to be transcriptionally arranged as nisA(Z)BTCIP, nisRK, and nisFEG. The biosynthesis of nisin is regulated in a growth-phase-dependent manner including nisin-mediated induction which occurs via NisRK two-component regulatory system. This review outlines some of the more recent developments in the properties, regulation and applications of nisin biosynthesis. PMID- 16247669 TI - A new method for mercury removal. AB - A method is described for the removal of mercury from solution by using the off gas produced from aerobic cultures of Klebsiella pneumoniae M426. Cells growing in Hg-supplemented medium produced a black precipitate containing mercury and sulphur. The ratio of Hg:S was determined as approximately 1:1 by analysis using proton-induced X-ray emission, suggesting precipitation of HgS within the culture. The outlet gases produced by a mercury-unsupplemented aerated culture were bubbled into an external chamber supplemented with up to 10 mg HgCl(2)/ml. A yellowish-white precipitate formed, corresponding to 99% removal of the mercury from solution within 120 min. Energy dispersive X-ray microanalysis showed that this metal precipitate consisted of mercury, carbon and sulphur. Formation of mercury carbonate was discounted since similar precipitation occurred at pH 2 and no oxygen was detected in the solid, which gave an X-ray powder pattern suggesting an amorphous material, with no evidence of HgS. Precipitation of mercury with a volatile organosulphur compound is suggested. Bio-precipitation of heavy metals by using culture off-gas is a useful approach because it can be used with concentrated or physiologically incompatible solutions. Since the metal precipitate is kept separate from the bacterial biomass, it can be managed independently. PMID- 16247670 TI - Mercury tolerance of thermophilic Bacillus sp. and Ureibacillus sp. AB - Although resistance of microorganisms to Hg(II) salts has been widely investigated and resistant strains have been reported from many eubacterial genera, there are few reports of mercuric ion resistance in extremophilic microorganisms. Moderately thermophilic mercury resistant bacteria were selected by growth at 62 degrees C on Luria agar containing HgCl(2). Sequence analysis of 16S rRNA genes of two isolates showed the closest matches to be with Bacillus pallidus and Ureibacillus thermosphaericus. Minimum inhibitory concentration (MIC) values for HgCl(2) were 80 microg/ml and 30 microg/ml for these isolates, respectively, compared to 10 microg/ml for B. pallidus H12 DSM3670, a mercury sensitive control. The best-characterised mercury-resistant Bacillus strain, B. cereus RC607, had an MIC of 60 microg/ml. The new isolates had negligible mercuric reductase activity but removed Hg from the medium by the formation of a black precipitate, identified as HgS by X-ray powder diffraction analysis. No volatile H(2)S was detected in the headspace of cultures in the absence or presence of Hg(2+), and it is suggested that a new mechanism of Hg tolerance, based on the production of non-volatile thiol species, may have potential for decontamination of solutions containing Hg(2+) without production of toxic volatile H(2)S. PMID- 16247671 TI - Predicting aggregation kinetics of DU 145 prostate cancer cells in liquid-overlay culture. AB - The predictive capacity of a novel population-balance model to simulate aggregation kinetics of attachment-dependent cells at the resolution of one-cell increments has been evaluated. Using spheroid assembly of DU 145 human prostate cancer cells as a representative system, the mathematical model proved to be robust in simulating aggregation over a 5-fold range of surface densities from 5 x 10(3) to 2.5 x 10(4) cells/cm(2) with a single matrix of rate constants. For cultures at 1 x 10(5) cells/cm(2), more than 75% of simulated aggregate concentrations are within the standard deviation of measured concentrations. For the two extreme densities, at least two-thirds of model predictions are within 35% of the mean for experimental data. Error in model predictions is attributed to uncertainty in measurements and intrinsic changes in aggregation. The model has application to the rational design of spheroids in tissue engineering and bioseparation processes in pharmaceutical manufacturing. PMID- 16247672 TI - A novel mucosal vaccine against foot-and-mouth disease virus induces protection in mice and swine. AB - Epitopes of a foot-and-mouth disease virus (FMDV) capsid protein VP1 complex and a chimera of 6xHis-tagged cholera toxin B subunit (hCTB) were expressed in Hansenula polymorpha and used together as a mucosal vaccine. Antibody and cytokine responses to VP1-hCTB vaccine and protection against FMDV were evaluated by ELISA and a virus challenge test in mice, respectively. VP1-hCTB directly enhanced the expression of interleukin-5 (IL-5) both in serum and supernatants of cultured spleen cells. After challenging suckling mice with 10(5) FMDV (=50% lethal dosage per mouse) a greater protection was seen after intraperitoneal and intranasal vaccinations than after oral vaccination. In swine immunized with VP1 hCTB, immune responses were achieved after three administrations, and the vaccine protected swine (80%) when challenged with 10(6.5) FMDV (=50% infectious dosage per swine). These results demonstrated the possibility of using CTB as a mucosal adjuvant to elicit protective immune responses against FMDV. PMID- 16247673 TI - Use of xylanase and arabinofuranosidase for arabinose removal from unbleached kraft pulp. AB - Preparations of arabinofuranosidase and xylanase, respectively from Aureobasidium pullulans and Trichoderma longibrachiatum, were used to remove selectively xylose and arabinose from kraft pulp. The equilibrium moisture content of pulps treated with both enzymes, at varying relative humidities, revealed a consistently lower percent moisture content at all humidity set points. Shorter fiber lengths indicated some deterioration when pulp was exposed to high concentrations of both enzymes. PMID- 16247674 TI - Expression of rice glutamate decarboxylase in Bifidobacterium longum enhances gamma-aminobutyric acid production. AB - Bifidobacteria are important for the production of fermented dairy products and probiotic formulas but have a low capacity for gamma-aminobutyric acid (GABA) production. To develop a Bifidobacterium strain with an enhanced GABA production, we transformed Bifidobacterium longum with a rice glutamate decarboxylase (OsGADC(-)) gene by electroporation. When the transformed strain was cultured in medium containing monosodium glutamate, the amount of GABA increased significantly compared with those of untransformed Bifidobacterium. Thus, by introducing a plant derived GAD gene, a Bifidobacterium strain has been genetically engineered to produce high levels of GABA from glutamate. PMID- 16247675 TI - L(+)-lactic acid production using Lactobacillus casei in solid-state fermentation. AB - Lactobacillus casei was grown at 37 degrees C on sugarcane bagasse (5 g) soaked with cassava starch hydrolysate (final moistening volume 34 ml) containing 3 g reducing sugar in a solid-state condition. The maximum yield of L-lactic acid after various process optimisations was 2.9 g/5 g initial substrate corresponding to 97% conversion of sugar to lactic acid with initial substrate moisture of 72%. PMID- 16247676 TI - Development of a biosensor for measuring plasma fibrinogen based on an enzyme kinetic model. AB - A biosensor has been designed for measuring plasma fibrinogen concentrations based on an enzyme kinetic model. The values obtained correspond linearly to those of a clinical method but have a higher sensitivity within the normal range of fibrinogen. By using the disposable biosensor technology, this method is easy to follow, quick (within 2 min) and precise (with coefficients of variation to be as low as below 5%) and requires only a small amount of plasma sample (<50 microl). This biosensor technology also holds potential for use in point-of-care diagnostics. PMID- 16247677 TI - Production of recombinant human placental variant growth hormone in Pichia pastoris. AB - cDNA encoding mature human placental variant growth hormone (HGH-V) was synthesized by retro-transcription polymerase chain reaction (RT-PCR) from total RNA recovered from human term-placenta and cloned in pBluescript plasmid (pBS) in Escherichia coli. cDNA was subcloned into pPIC9, fusing it to the flanking regulatory sequences of the Pichia pastoris alcohol oxidase 1 gene (AOX1) and finally introduced into the genome of this yeast by homologous recombination. The resulting new recombinant strain produced and secreted, towards the culture medium, mature HGH-V, whose activity was demonstrated in cell culture by the Nb2 proliferation assay. PMID- 16247678 TI - Assembly of plasmid DNA into liposomes after condensation by cationic lipid in anionic detergent solution. AB - A novel strategy to prepare negatively charged and small DNA-containing liposomes after condensation of plasmid DNA by a cationic lipid in deoxycholate micelle environment is described. The average diameter of resulting complexes was 62 +/- 8 nm. DNA-containing liposomes were then prepared by dialysis. The shape of the resulting liposomes was spherical. The average diameter and the surface charge of the liposomes were 86 +/- 6 nm and -24 +/- 3 mV, respectively. The plasmid DNA inside liposomes remained in a supercoiled form after incubation with DNase. PMID- 16247679 TI - Development of a plasmid display system using GAL4 DNA binding domain for the in vitro screening of functional proteins. AB - A plasmid display system using GAL4 DNA binding domain (GAL4 DBD) was constructed to enrich the molecular diversity and in vitro selection of functional proteins. Model proteins used were enhanced green fluorescent protein (EGFP) and glutathione S-transferase (GST). The feasibility of this display system was examined using enrichment experiments of target protein from a model protein mixture and identifying the encoding genes by PCR, in which the model protein mixture includes GAL4 DBD/GST fusion protein, GAL4 DBD/EGFP fusion protein, and xylanase. Target proteins of GAL4 DBD/GST and GAL4 DBD/EGFP from the model protein mixture were efficiently isolated by the plasmid display, respectively. The results show that the display system is sufficiently sensitive to select a target protein from a protein mixture, and that it is possible to discover the functional proteins from large libraries using relatively simple approaches. PMID- 16247680 TI - Expression of chimeric antibody in mammalian cells using dicistronic expression vector. AB - A dicistronic expression vector was constructed for Chinese hamster ovary (CHO) cells that produce both selectable marker-DHFR (dihydrofolate reductase) gene and recombinant antibody cDNA from a single primary transcript via differential splicing. The vector was derived from a pDHL vector and contained the human constant region cDNA so that any human-mouse chimeric antibodies could be expressed. The expression vector produced stable CHO cell clones that secreted nearly double the amount of chimeric antibodies than produced by conventional expression approaches, where the DHFR gene and relevant cDNA are controlled by separate transcription cassettes. Clones with increased expression of interested genes can be efficiently generated by selection in medium containing a gradually increasing amount of methotrexate. The dicistronic expression system using incomplete splicing DHFR gene strategy thus provides a convenient, high-level, and rapid expression of chimeric antibodies. PMID- 16247682 TI - Culture of isolated single cells from Taxus suspensions for the propagation of superior cell populations. AB - Single cells isolated from aggregated Taxus cuspidata cultures via enzymatic digestion were grown in suspension culture. High seeding density (4 x 10(5 )cells/ml) and the addition of cell-free conditioned medium were essential for growth. Doubling the concentration of the nutrients [ascorbic acid (150 g/l), glutamine (6.25 mM: ), and citric acid (150 g/l)] had no effect on single cell growth or viability. A specific growth rate of 0.11 days(-1) was achieved, which is similar to the observed growth rate of aggregated Taxus suspensions. The biocide, Plant Preservative Mixture, added at 0.2% (v/v) to all single cell cultures to prevent microbial contamination, had no significant effect on growth or viability. Following cell sorting, single cell cultures can be used to establish new cell lines for biotechnology applications or provide cells for further study. PMID- 16247681 TI - Production of deuterated zeaxanthin by Flavobacterium multivorum and its detection by resonance Raman and mass spectrometric methods. AB - Flavobacterium multivorum, a zeaxanthin-producing organism, was grown aerobically in a medium prepared with deuterated water. Atmospheric pressure chemical ionization mass spectrometry (APCI-MS) and resonance Raman spectroscopy (RRS) analysis revealed approximately 75% replacement of hydrogen by deuterium atoms as indicated by the molecular mass cluster at around m/z 600. Deuterated zeaxanthin upon excitation with a 488 nm laser exhibited characteristic resonance Raman vibrational modes at 1,161 and 1,504 cm(-1) as compared to 1,007, 1,159 and 1,525 cm(-1 )for undeuterated zeaxanthin. HPLC/APCI-MS and HPLC/RRS were specific and sensitive with limits of detection of 2.5 pg and 50 ng, respectively. PMID- 16247685 TI - Genetic structure of wild rice Oryza glumaepatula populations in three Brazilian biomes using microsatellite markers. AB - The existence of Oryza glumaepatula is threatened by devastation and, thus, the implementation of conservation strategies is extremely relevant. This study aimed to characterize the genetic variability and estimate population parameters of 30 O. glumaepatula populations from three Brazilian biomes using 10 microsatellite markers. The levels of allelic variability for the SSR loci presented a mean of 10.3 alleles per locus and a value of 0.10 for the average allelic frequency value. The expected total heterozygosity (H(e)) ranged from 0.63 to 0.86. For the 30 populations tested, the mean observed (H(o)) and expected heterozygosities (H(e)) were 0.03 and 0.11 within population, respectively, indicating an excess of homozygotes resulting from the preferentially self-pollinating reproduction habit. The estimated fixation index ( (IS) ) was 0.79 that differed significantly from zero, indicating high inbreeding within each O. glumaepatula population. The total inbreeding of the species ((IT) ) was 0.98 and the genetic diversity indexes among populations, (ST) and (ST), were 0.85 and 0.90, respectively, indicating high genetic variability among them. Thus, especially for populations located in regions threatened with devastation, it is urgent that in situ preservation conditions should be created or that collections be made for ex situ preservation to prevent loss of the species genetic variability. PMID- 16247683 TI - Expression and secretion of an acid-stable alpha-amylase gene in Bacillus Subtilis by SacB promoter and signal peptide. AB - Alpha amylase gene from Bacillus licheniformis was mutated by site-directed mutagenesis to improve its acid stability. The mutant gene was expression in Bacillus subtilis under the control of the promoter of sacB gene which was followed by either the alpha-amylase leader peptide of Bacillus licheniformis or the signal peptide sequence of sacB gene of Bacillus subtilis. Both peptides efficiently directed the secretion of alpha-amylase from the recombinant B. subtilis cells. The extracellular alpha-amylase activities in two recombinants were 1001 and 2012 U ml(-1), respectively. The purity of the recombinant product was confirmed by SDS-PAGE. PMID- 16247686 TI - Chromosome banding and 18S rDNA in situ hybridization analysis of seven species of the family Achiridae (Teleostei: Pleuronectiformes). AB - Achiridae is an important family of the order Pleuronectiformes widely distributed in North, Central, and South America with freshwater and marine species. In the present study cytogenetic analyses comprising conventional and molecular techniques were carried out in seven species of this family. The following diploid numbers (2n) and fundamental numbers (FN) were obtained: Achirus declivis 2n=34, FN=52; Achirus lineatus 2n=40, FN=66; Catathyridium jenynsi 2n=40 and FN=50; Gymnachirus nudus 2n=36 and FN=50; Hypoclinemus mentalis 2n=38 and FN=54; Trinectes paulistanus 2n=42 and FN=52; and Trinectes sp. 2n=38 and FN=54. All species presented a single nucleolar organizer region (NOR) bearing chromosome pair and C-band positive segments mainly distributed at the pericentromeric position. The wide variation observed in chromosome number and FN suggests the occurrence of larger chromosome rearrangements in the family Achiridae if compared with other families of the same order. PMID- 16247687 TI - Genetic diversity among Paspalum L. species (Poaceae) belonging to the Notata and Linearia groups based on restriction fragment length polymorphism analyses. AB - A taxonomic study of Paspalum L. was carried out using a genetic diversity approach. Thirty accessions representing twenty one different species from the Notata and Linearia groups of Paspalum were studied using restriction fragment length polymorphism analysis of the amplified ITS ribosomal DNA (rDNA) and from the psbA-trnH of the chloroplast genome (cpDNA). The combined analysis of the internal transcribed spacer (ITS) and the chloroplast spacer region between the psbA and trnH genes identified genetic polymorphisms. A distance analysis of the molecular data generated a dendrogram which showed the relationships of the two informal groups of Paspalum studied here. Although the distribution of species in the dendrogram was found to be roughly in agreement with previous works based on morphological and cytological data, the results obtained reveal the current artificiality in Paspalum taxonomy. Based on molecular data, a new circumscription of the Notata and Linearia groups is proposed here in order to provide a more accurate delimitation of these groups and contribute to the taxonomy of Paspalum. This study, although preliminary, reveals the potential utility of such a molecular approach for clarifying the taxonomy of closely related taxa. PMID- 16247688 TI - Intraspecific phylogeography of red squirrels (Tamiasciurus hudsonicus) in the central Rocky Mountain region of North America. AB - We used variation in a portion of the mitochondrial DNA control region to examine phylogeography of Tamiasciurus hudsonicus, a boreal-adapted small mammal in the central Rocky Mountain region. AMOVA revealed that 65.66% of genetic diversity was attributable to variation within populations, 16.93% to variation among populations on different mountain ranges, and 17.41% to variation among populations within mountain ranges. Nested clade analysis revealed two major clades that likely diverged in allopatry during the Pleistocene: a southern clade from southern Colorado and a northern clade comprising northern Colorado, Wyoming, eastern Utah, and eastern Idaho. Historically restricted gene flow as a result of geographic barriers was indicated between populations on opposite sides of the Green River and Wyoming Basin and among populations in eastern Wyoming. In some instances genetic structure indicated isolation by distance. PMID- 16247689 TI - Population genetics and identity of an introduced terrestrial slug: Arion subfuscus s.l. in the North-East USA (Gastropoda, Pulmonata, Arionidae). AB - Several European species of the terrestrial slug genus Arion have been introduced into North America. A case in point is the species complex A. subfuscus s.l. which has become one of the most abundant slug taxa in North America. In Europe this complex consists of at least two cryptic species, viz. A. fuscus and A. subfuscus s.s., the latter of which is further subdivided in five strongly divergent mtDNA lineages (A. subfuscus S1-S5). In order to determine which of these A. subfsucus s.l. taxa are present in the NE USA and in order to assess their population genetic structure, we compared mtDNA, nDNA and allozyme variation between populations from the NE USA and Europe. Our results show that (1) at least A. subfuscus S1 has become successfully established in the NE USA, (2) founder effects are the most likely explanation for the loss of a large amount of molecular genetic variation in populations from the NE USA (i.e. a loss of 96% of the 16S rDNA haplotypes, 67% of the ITS1 alleles and 46% of the alleles at polymorphic allozyme loci), and (3) part of the remaining genetic variation in NE USA populations was probably due to multiple introductions from the British Isles and the European mainland, and the hybrid structure of most of these source populations. Apparently, the extreme loss of molecular genetic variation in this introduced species has not prevented it from successfully establishing and spreading in novel environments. PMID- 16247691 TI - Rate of chromosome changes and speciation in reptiles. AB - The chromosome changing rate (i.e. the number of chromosome rearrangements per million years) was studied in 1,329 reptile species in order to evaluate the karyological evolutionary trend and the existence of possible correlations between chromosome mutations and some aspects of the evolution of this class. The results obtained highlight the existence of a general direct correlation between chromosome changing rate and number of living species, although different trends can be observed in the different orders and suborders. In turtles, the separation of pleurodires from cryptodires was accompanied by a considerable karyological diversification. Among pleurodires, the evolution of the Chelidae and Pelomedusidae was also characterised by chromosome variation, while in cryptodires a marked karyological homogeneity is observed between and within infra-orders. Similarly there is no correlation between changing rate and species number in crocodiles, where the evolution of the families and genera has entailed few chromosome mutations. Chromosome variability was greater in lizards and snakes. In the formers variations in chromosome changing rate accompanied the separation of the infra-orders and the evolution of most of the families and of some genera. The origin of snakes has also been accompanied by a marked karyological diversification, while the subsequent evolution of the infra-orders and families has entailed a high level of chromosome variability only in colubroids. The karyological evolution in reptiles generally entailed a progressive reduction in chromosome changing rate, albeit with differences in the diverse orders and suborders. This trend seems to be consistent with the "canalization model" as originally proposed by Bickham and Baker in [Bickham, J.W. & R J. Baker, 1979. Bull. Carnegie Mus. Nat. Hist. 13: 70-84.] However, several inconsistencies have been found excluding that in this class the ultimate goal of chromosome variations was the achievement of a so-called "optimum karyotype'' as suggested by the above-mentioned theory. Other mechanisms could underpin chromosome variability in Reptiles. Among them a genomic composition more or less favourable to promoting chromosome rearrangements and factors favouring the fixation of a mutant karyotype in condition of homozygosis. Turtles and crocodiles would have a genome characterised by large chromosomes and a low level of chromosome compartmentalisation limiting the recombination and the frequency of rearrangements. A low rate of chromosome variability modifying little if at all the gene linkage groups would have favoured a conservative evolutionary strategy. In the course of evolution, lizards and snakes could have achieved a genome characterised by smaller chromosomes and a higher level of compartmentalisation. This would have raised the frequency of recombination and consequently an evolutionary strategy promoting a higher degree of variability and a greater level of speciation. PMID- 16247690 TI - Genetic variability and genetic structure of wild and semi-domestic populations of tasar silkworm (Antheraea mylitta ) ecorace Daba as revealed through ISSR markers. AB - The genetic diversity in the wild and semi-domestic populations of Daba ecorace of Antheraea mylitta was studied to ascertain the distribution of variability within and among populations of semi-domestic bivoltine (DB), trivoltine (DT) and nature grown wild populations (DN) with inter-simple sequence repeat (ISSR) markers. A total of 138 markers were produced among 56 individuals of the three populations, of which 98% were polymorphic. For the individual populations, the percentage polymorphism was 58.69, 52.9 and 77.54 for DB, DT and DN, respectively. Average number of observed (1.791+/- 0.408) and effective alleles (1.389+/-0.348) was also high in the wild populations in comparison to the bivoltine and trivoltine semi-domestic populations. Genetic diversity (H(t)) in DB, DT and DN was 0.180+/- 0.033, 0.153+/- 0.032 and 0.235+/- 0.033, respectively and within-population genetic diversity (H(s)) ranged from 0.166 to 0.259 with a mean of 0.189. Mean gene differentiation (G(ST)) was found to be 0.25. Shanon's diversity index was 0.278, 0.237 and 0.361 for DB, DT and DN and overall it was 0.391. Gene flow (N(m)) among the populations was 1.509. The dendrogram produced by UPGMA with Dice's genetic distance matrices resulted in the formation of three major clusters separating the three populations. Considerable intra- and inter population variability is found in all three populations. The population structure analysis further suggests that the semi-domestic populations of Daba ecorace are at the threshold of differentiating themselves. The high genetic variability present within wild Daba population of A. mylitta is of much importance for conservation as well as utilization in systematic breeding program. PMID- 16247692 TI - Isolation and characterization of a satellite DNA family in Achirus lineatus (Teleostei: Pleuronectiformes: Achiridae). AB - Agarose gels stained with Ethidium bromide and Southern blot experiments of HindIII-digested genomic DNA of Achirus lineatus evidenced the presence of monomers and multimers of a DNA segment of about 200 bp, named here Al-HindIII sequence. No signals were observed in Southern blot experiments with genomic DNA of other flatfish species. The DNA sequencing of four recombinant clones showed that Al-HindIII sequences had 204 bp and were 63.72% AT-rich. FISH experiments using a Al-HindIII sequence as probe showed bright signals in the centromeric position of all chromosomes of A. lineatus. PMID- 16247694 TI - Identification of one intron loss and phylogenetic evolution of Dfak gene in the Drosophila melanogaster species group. AB - Intron loss and its evolutionary significance have been noted in Drosophila. The current study provides another example of intron loss within a single-copy Dfak gene in Drosophila. By using polymerase chain reaction (PCR), we amplified about 1.3 kb fragment spanning intron 5-10, located in the position of Tyr kinase (TyK) domain of Dfak gene from Drosophila melanogaster species group, and observed size difference among the amplified DNA fragments from different species. Further sequencing analysis revealed that D. melanogaster and D. simulans deleted an about 60 bp of DNA fragment relative to other 7 Drosophila species, such as D. elegans, D. ficusphila, D. biarmipes, D. takahashii, D. jambulina, D. prostipennis and D. pseudoobscura, and the deleted fragment located precisely in the position of one intron. The data suggested that intron loss might have occurred in the Dfak gene evolutionary process of D. melanogaster and D. simulans of Drosophila melanogaster species group. In addition, the constructed phylogenetic tree based on the Dfak TyK domains clearly revealed the evolutionary relationships between subgroups of Drosophila melanogaster species group, and the intron loss identified from D. melanogaster and D. simulans provides a unique diagnostic tool for taxonomic classification of the melanogaster subgroup from other group of genus Drosophila. PMID- 16247693 TI - Genetic architecture of two fitness-related traits in Drosophila melanogaster: ovariole number and thorax length. AB - In Drosophila melanogaster, ovariole number and thorax length are morphological characters thought to be associated with fitness. Maximum daily egg production in females is positively correlated with ovariole number, while thorax length is correlated with male reproductive success and female fecundity. Though both traits are related to fitness, ovariole number is likely to be under stabilizing selection, while thorax length appears to be under directional selection. Current research has focused on examining the sources of variation for ovariole number in relation to fitness, with a view towards elucidating how segregating variation is maintained in natural populations. Here, we utilize a diallel design to explore the genetic architecture of ovariole number and thorax length in nine isogenic lines derived from a natural population. The full diallel design allows the estimation of general combining ability (GCA), specific combining ability (SCA), and also describes variation due to reciprocal effects (RGCA and RSCA). Ovariole number and thorax length differed with respect to their genetic architecture, reflective of the independent selective forces acting on the traits. For ovariole number, GCA accounted for the majority (67.3%) of variation segregating between the lines, with no evidence of reciprocal effects or inbreeding depression; SCA accounted for a small percentage (3.9%) of the variance, suggesting dominance variation; no reciprocal effects were observed. In contrast, for thorax length, the majority of the non-error variance was accounted for by SCA (17.9%), with only one third as much variance (6.2%) due to GCA. Interestingly, RSCA (nuclear extranuclear interactions) accounted for slightly more variation (7.5%) than GCA in these data. Thus, genetic variation for thorax length is largely in accord with predictions for a fitness trait under directional selection: little additive genetic variation and substantial dominance variation (including a suggestion of inbreeding depression); while the mechanisms underlying the maintenance of variation for ovariole number are more complex. PMID- 16247695 TI - Microsatellite analysis of olive fly populations in the Mediterranean indicates a westward expansion of the species. AB - Bactrocera oleae is the major insect pest of the olive fruit. Twelve microsatellite loci isolated from the genome of this insect were used in a Mediterranean-wide population analysis. These loci were highly polymorphic with a mean number of alleles per locus of 10.42 and a mean effective number of alleles of 2.76. The analysis was performed on a sample of 671 flies collected from nineteen locations around the European part of the Mediterranean basin. Despite the high level of gene flow across the Mediterranean, results support the notion of a differentiation of three subpopulations: one of the Iberian Peninsula, one of Greece and Italy and one of Cyprus. In addition, the gradual decrease of heterozygosity from the Eastern to the Western part of the Mediterranean indicates a westward expansion of the species. PMID- 16247696 TI - Identification of novel non-autonomous CemaT transposable elements and evidence of their mobility within the C. elegans genome. AB - We describe here two new transposable elements, CemaT4 and CemaT5, that were identified within the sequenced genome of Caenorhabditis elegans using homology based searches. Five variants of CemaT4 were found, all non-autonomous and sharing 26 bp inverted terminal repeats (ITRs) and segments (152-367 bp) of sequence with similarity to the CemaT1 transposon of C. elegans. Sixteen copies of a short, 30 bp repetitive sequence, comprised entirely of an inverted repeat of the first 15 bp of CemaT4's ITR, were also found, each flanked by TA dinucleotide duplications, which are hallmarks of target site duplications of mariner-Tc transposon transpositions. The CemaT5 transposable element had no similarity to maT elements, except for sharing identical ITR sequences with CemaT3. We provide evidence that CemaT5 and CemaT3 are capable of excising from the C. elegans genome, despite neither transposon being capable of encoding a functional transposase enzyme. Presumably, these two transposons are cross mobilised by an autonomous transposon that recognises their shared ITRs. The excisions of these and other non-autonomous elements may provide opportunities for abortive gap repair to create internal deletions and/or insert novel sequence within these transposons. The influence of non-autonomous element mobility and structural diversity on genome variation is discussed. PMID- 16247697 TI - A comparative cytogenetic analysis between the grasshopper species Chromacris nuptialis and C. speciosa (Romaleidae): constitutive heterochromatin variability and rDNA sites. AB - The chromosomes of Chromacris nuptialis and C. speciosa were comparatively analyzed using different cytogenetic techniques, in order to determine the level of karyotypic similarities and differences between the species. The results show similarities in chromosome number (2n=23,X0) and acrocentric morphology. In some C. nuptialis individuals meiotic irregularities were detected involving the L(2) bivalent. This bivalent was delayed and presented anaphasic bridges and other aberrations. Differences in constitutive heterochromatin (CH) patterns and composition were observed through C-banding and fluorochromes staining. Silver nitrate staining revealed a single medium nucleolar organizer regions (NORs) pair, per species. Differences were also observed in NORs location, which was pericentromeric in C. nuptialis and proximal in C. speciosa. FISH using an rDNA probe confirmed the existence of ribosomal sites coinciding with active regions visualized by silver nitrate. The possible implications of the karyotype differences observed between both species are discussed. PMID- 16247698 TI - Cytogenetic analysis of experimental hybrids in species of Triatominae (Hemiptera Reduviidae). AB - A cytogenetic analysis was performed in experimental hybrids between species of Chagas disease transmitting bugs with remarkable differences in the amount and distribution of heterochromatin. Using C-banding technique, we identified the parental species chromosomes and analysed the meiotic behaviour in the male hybrids between Triatoma platensis and T. infestans, T. platensis and T. delpontei, and T. infestans and T. rubrovaria. The two former hybrids have an entirely normal meiotic behaviour despite the extensive differences in C-banded karyotypes observed in the parental species, indicating that heterochromatin differences between homeologous chromosomes are not a barrier that influences meiotic synapsis and recombination. On the contrary, the experimental hybrids between T. infestans and T. rubrovaria show failures in pairing of homeologous chromosomes that lead to the production of abnormal spermatids and hybrid sterility. Our data suggest that karyotypic repatterning within triatomines has involved at least two different pathways. Among closely related species, chromosomal changes have largely involved addition or deletion of heterochromatic regions. In more distant species, chromosomal rearrangements (i.e. inversions and translocations) have also arisen. Hybridisation data also allow to hypothesize about the origin and divergence of this taxonomic group, as well as the mechanisms that maintain species isolation. PMID- 16247699 TI - Variability of wing size and shape in three populations of a recent Brazilian invader, Zaprionus indianus (Diptera: Drosophilidae), from different habitats. AB - Zaprionus indianus (Diptera: Drosophilidae) is an African species that was introduced in Brazil near the end of the 1990's decade. To evaluate the adaptive potential of morphological traits in natural populations of this recently introduced species, we have investigated wing size and shape variation at Rio de Janeiro populations only two years after the first record of Z. indianus in Brazil. Significant genetic differences among populations from three distinct ecological habitats were detected. The heritability and evolvability estimates show that, even with the population bottleneck that should have occurred during the invasion event, an appreciable amount of additive genetic variation for wing size and shape was retained. Our results also indicated a greater influence of environmental variation on wing size than on wing shape. The importance of quantitative genetic variability and plasticity in the successful establishment and dispersal of Z. indianus in the Brazilian territory is then discussed. PMID- 16247700 TI - Four tropical, closely related fern species belonging to the genus Adiantum L. are genetically distinct as revealed by ISSR fingerprinting. AB - The level and pattern of genetic variation was analyzed in four species of the fern genus Adiantum L., A. hispidulum Sw., A. incisum Forrsk., A. raddianum C.Presl, and A. zollingeri Mett. ex Kuhn, originating from South India, using the ISSR fingerprinting method. The populations of Adiantum possessed a considerable level of genetic variation, the diversity indices ranging from 0.284 to 0.464. Only 12% of the ISSR markers found were restricted to one species only, and 54% were detected in all four species. The analysis of molecular variance revealed that 71.1% of variation was present within populations. The proportion of variation detected among species was only 18.5% while the proportion of variation among populations within species equalled 10.4%. Despite the low level of intrageneric differentiation, the discriminant analysis and clustering of genetic distances indicated that the four Adiantum species are genetically distinct. The F(ST) values calculated for the species were low, varying from 0.089 to 0.179. No linkage disequilibrium was detected between the loci. Such low level of differentiation among populations and the presence of linkage equilibrium reflect that the life history of Adiantum ferns apparently involves common or relatively common sexuality, effective wind-dispersal of spores and outcrossing. PMID- 16247701 TI - A second look at mitochondrial DNA variability in European anchovy (Engraulis encrasicolus): assessing models of population structure and the Black Sea isolation hypothesis. AB - Genetic architectures of marine fishes are generally shallow because of the large potential for gene flow in the sea. European anchovy, however, are unusual among small pelagic fishes in showing large differences among sub-basins and in harbouring two mtDNA phylogroups ('A' & 'B'), representing 1.1-1.85 million years of separation. Here the mtDNA RFLP dataset of Magoulas et al. [1996, Mol. Biol. Evol. 13: 178-190] is re-examined to assess population models accounting for this subdivided population structure and to evaluate the zoogeographical origins of the two major phylogroups. Haplotype and nucleotide diversities are highest in the Ionian Sea and lowest in the Aegean and Black seas. However, this gradient is absent when 'A' and 'B' haplotypes are examined separately. Neither the self sustaining nor the basin population models adequately describe anchovy population behaviour. Tests for neutrality, mismatch and nested clade analyses are concordant in depicting recent expansions of both phylogroups. Unimodel mismatch distributions and haplotype coalescences dating to the last (Eemian) interglacial ('B') and the Weichselian pleniglacial period ('A') indicate separate colonizations of the Mediterranean Basin. Phylogroup 'A' is unlikely to have arisen through continuous long-term isolation in the Black Sea because of climate extremes from displaced subpolar weather systems during the ice ages. Ancestors of both groups appear to have colonized the Mediterranean from the Atlantic in the late Pleistocene. Hence, zoogeographic models of anchovy in the Mediterranean must also include the eastern (and possibly southern) Atlantic. PMID- 16247702 TI - Population genetic structure of two columnar cacti with a patchy distribution in eastern Brazil. AB - The genetic variability and population genetic structure of six populations of Praecereus euchlorus and Pilosocereus machrisii were investigated. The genetic variability in single populations of Pilosocereus vilaboensis, Pilosocereus aureispinus, and Facheiroa squamosa was also examined. All of these cacti species have a patchy geographic distribution in which they are restricted to small areas of xeric habitats in eastern Brazil. An analysis of genetic structure was used to gain insights into the historical mechanisms responsible for the patchy distribution of P. euchlorus and P. machrisii. High genetic variability was found at the populational level in all species (P=58.9-92.8%, A(p)=2.34-3.33, H(e)=0.266-0.401), and did not support our expectations of low variability based on the small population size. Substantial inbreeding was detected within populations (F(IS)=0.370-0.623). In agreement with their insular distribution patterns, P. euchlorus and P. machrisii had a high genetic differentiation (F(ST)=0.484 and F(ST)=0.281, respectively), with no evidence of isolation by distance. Accordingly, estimates of gene flow (N(m)) calculated from F(ST) and private alleles were below the level of N(m)=1 in P. machrisii and P. euchlorus. These results favored historical fragmentation as the mechanism responsible for the patchy distribution of these two species. The genetic distance between P. machrisii and P. vilaboensis was not compatible with their taxonomic distinction, indicating a possible local speciation event in this genus, or the occurrence of introgression events. PMID- 16247704 TI - The fundamental theorem of neutral evolution: rates of substitution and mutation should factor in premeiotic clusters. AB - Mutations do not always arise as single events. Many new mutations actually occur in the cell lineage before germ cell formation or meiosis and are therefore replicated pre-meiotically. The increased likelihood of substitutions caused by these clusters of new mutant alleles can change the fundamental theorem of neutral evolution. PMID- 16247703 TI - Advent of a new retrotransposon structure: the long form of the Veju elements. AB - Transposable elements are the main component of plant genomes, especially in grass species. In a previous analysis, we have identified two unusual types of Class I elements, two homologous Veju TRIM elements, but with an unusual long structure. They are formed by the junction of a yet unidentified segment labelled unknown DNA, flanked by the borders of the classical Veju element. Here, we show that the long (Veju_L) and the short forms (Veju_S) coexist within wheat genomes. The associated unknown DNA had always the same origin, and the Veju_L came probably from either illegitimate recombinations or 'template switching' between the Veju_S and another unique unknown DNA sequence. This junction then evolved differently within wheat genomes. PMID- 16247707 TI - The prostacyclin analogue beraprost sodium prevents occlusion of bypass grafts in patients with lower extremity arterial occlusive disease: a 20-year retrospective study. AB - Although conventional bypass grafting is commonly used to treat ischemia in lower extremities, graft failure often occurs. This study retrospectively analyzed the factors that affect graft patency to help establish more effective treatment of obstructive arterial disease of the lower limbs. Kaplan-Meier analysis was used to estimate graft patency in 90 legs of 80 patients who underwent femoropopliteal bypass (28 vein grafts and 62 expanded polytetrafluoroethylene grafts) between 1984 and 2003. Patients were randomly selected for graft materials in sequential surgical treatment order. After initial analysis, several risk factors and postoperative medication regimens were analyzed to ascertain any association with graft failure. The overall mean patency period for femoropopliteal bypass was 10.5 +/- 0.7 years. Graft occlusion occurred in 20 limbs. Neither the materials composing the grafts nor the position of distal anastmosis had any influence on patency maintenance. Graft occlusion rates were significantly greater in patients with either diabetes (p = 0.0049) or rest pain before surgery (p = 0.0011). Postoperative administration of beraprost sodium significantly increased the patency period (p = 0.0082). Diabetes and rest pain before surgery are important factors for late graft failure after femoropopliteal bypass. Our data also suggest that administration of beraprost sodium increases the graft patency period. PMID- 16247708 TI - Microcirculation and venous ulcers: a review. AB - Recent histological and immunocytochemical analyses of venous leg ulcers suggest that lesions observed in the different stages of chronic venous insufficiency (CVI) may be related to an inflammatory process. This inflammatory process leads to fibrosclerotic remodeling of the skin and then to ulceration. The vascular network of the most superficial layers of the skin appears to be the target of the inflammatory reaction. Hemodynamic forces such as venous hypertension, circulatory stasis, and modified conditions of shear stress appear to play an important role in an inflammatory reaction accompanied by leukocyte activation which clinically leads to CVI: venous dermatitis and venous ulceration. The leukocyte activation is accompanied by the expression of integrins and by synthesis and release of many inflammatory molecules, including proteolytic enzymes, leukotrienes, prostaglandin, bradykinin, free oxygen radicals, cytokines, and possibly other classes of inflammatory mediators. The inflammatory reaction perpetuates itself, leading to liposclerotic skin and subcutaneous tissue remodeling. In light of the mechanisms of venous ulcer formation cited above, therapy in the future might be directed against leukocyte activation in order to diminish the magnitude of the inflammatory response. With this in mind, the attention of many investigators has been drawn to two different drugs with an anti-inflammatory effect: pentoxifylline and flavonoids. PMID- 16247709 TI - BCRP transports dipyridamole and is inhibited by calcium channel blockers. AB - PURPOSE: We investigated whether dipyridamole and various calcium channel blockers are inhibitors and/or substrates of breast cancer resistance protein (BCRP). METHODS: The effect of dipyridamole and the calcium channel blockers on mitoxantrone efflux by BCRP-overexpressing human embryonic kidney (HEK) cells was determined by flow cytometry. The ability of some of these compounds to reverse BCRP-mediated mitoxantrone resistance was measured by cytotoxicity assays. Transport studies were performed using radiolabeled compounds. RESULTS: Dipyridamole, nicardipine, nitrendipine, and nimodipine effectively inhibited BCRP-mediated mitoxantrone efflux; however, bepridil, diltiazem, and verapamil had no significant effect. Nifedipine is a much weaker BCRP inhibitor compared with other dihydropyridines tested. Nicardipine and dipyridamole were the most potent BCRP inhibitors among the compounds tested with IC50 values of 4.8 +/- 1.3 and 6.4 +/- 0.9 microM, respectively. Nicardipine and dipyridamole also effectively reversed BCRP-mediated mitoxantrone resistance in HEK cells. [3H]Nitrendipine was found not to be transported by BCRP. However, the transport of [3H]dipyridamole by BCRP was observed in both HEK and Madin-Darby canine kidney cells stably expressing the transporter, and this transport was completely abolished by fumitremorgin C, a known BCRP inhibitor. CONCLUSIONS: Dipyridamole and several dihydropyridines are effective BCRP inhibitors, but bepridil, diltiazem, and verapamil are not. We also identified a new BCRP substrate, dipyridamole. PMID- 16247710 TI - Pharmacokinetic significance of renal OAT3 (SLC22A8) for anionic drug elimination in patients with mesangial proliferative glomerulonephritis. AB - PURPOSE: Our previous studies showed that the mRNA level of human organic anion transporter (hOAT) 3 in the kidney was correlated with the rate of elimination of an anionic antibiotic cefazolin. However, the correlation coefficient was not so high. In the present study, therefore, we enrolled more patients to examine whether additional factors were responsible for the correlation. METHODS: hOAT mRNA levels in renal biopsy specimens were quantified using the real-time polymerase chain reaction method. The elimination rates for the free fraction of cefazolin were determined in patients with various renal diseases. RESULTS: In the present study, the coefficient of correlation between the hOAT3 mRNA level and the elimination rates for the free fraction of cefazolin was not so high in the patients overall as in our previous study (r = 0.536). However, following the classification of renal diseases, a better correlation was obtained in patients with mesangial proliferative glomerulonephritis (r = 0.723). In contrast, multiple regression analyses including gender, age, and liver function did not result in any improvements in the correlation coefficients. CONCLUSIONS: These results suggest that the hOAT3 mRNA level is a significant marker of pharmacokinetics with which to predict the rate of elimination of cefazolin in patients with mesangial proliferative glomerulonephritis. PMID- 16247711 TI - The effects of food on the dissolution of poorly soluble drugs in human and in model small intestinal fluids. AB - PURPOSE: This study was conducted to determine the effect of food on drug solubility and dissolution rate in simulated and real human intestinal fluids (HIF). METHODS: Dissolution rate obtained via the rotating disk method and saturation solubility studies were carried out in fed and fasted state HIF, fed dog (DIF), and simulated (FeSSIF) intestinal fluid for six aprotic low solubility drugs. The intestinal fluids were characterized with respect to physical-chemical characteristics and contents. RESULTS: Fed HIF provided a 3.5- to 30-times higher solubility compared to fasted HIF and FeSSIF, whereas fed DIF corresponded well (difference of less than 30%) to fed HIF. The increased solubility of food could mainly be attributed to dietary lipids and bile acids. The dissolution rate was also 2 to 7 times higher in fed HIF than fasted HIF. This was well predicted by both DIF and FeSSIF (difference of less than 30%). CONCLUSIONS: Intestinal solubility is higher in fed state compared to fasted state. However, the dissolution rate does not increase to the same extent. Dog seems to be a good model for man with respect to dissolution in the small intestine after intake of a meal, whereas FeSSIF is a poorer means of determining intestinal saturation solubility in the fed state. PMID- 16247712 TI - Predicting human cloning acceptability: a national Greek survey on the beliefs of the public. AB - BACKGROUND: In the era of human genome research, there is a large theoretical debate among scientists and authorities on the ethical dimension based on the moral liberty of the individuals and the scientific and economic dimension based on the freedom and the independence of the scientific and technological activities. Meanwhile, the understanding of beliefs on human cloning (HC) and its acceptability are important for the development of evidence-based policy making. However, previous research in the field of public beliefs towards human genetics is limited. Therefore, in the present study, we investigated the role of public beliefs as predictors of the acceptability of the respondents to use HC. METHODS: Personal interviews were conducted with 1020 men and women of urban areas in Greece. Stratified random sampling was performed to select participants. Several scientists, experts in HC, evaluated the content of the instrument initially developed. The final questionnaire was the result of a pilot study. RESULTS: The acceptability of HC for the cure of incurable diseases and transplantation need is very high (70.7 and 58.6%, respectively). Public's intention to have recourse to HC because of "bringing" back to life a loved person or because of reproductive disorders was reported by 35 and 32.5%, respectively. With respect to the role of beliefs: increasing scores of reasons of social benefits, moral/religious reasons and legislative reasons increased the public's intention to have recourse to HC; inversely, decreasing scores of reasons of human commodification/exploitation increased public's intention to have recourse to HC. Additionally, low rates of church attendance appeared to be correlated with high reported acceptability of HC. CONCLUSION: There is great public concern regarding the application of HC, which probably reflects the existing ambivalence over the relationship between technology and society. Scientists and policymakers should take into account these indicators of public disquiet and should manage the public involvement in policy decisions, from which they have so far been excluded. PMID- 16247714 TI - Endometrioma undergoing laparoscopic ovarian cystectomy: its influence on the outcome of in vitro fertilization and embryo transfer (IVF-ET). AB - PURPOSE: To evaluate the effect of laparoscopic ovarian cystectomy for endometrioma on the clinical outcome of IVF treatment. METHODS: Patients who received IVF treatment were retrospectively classified into two groups. Group 1 included 95 patients who received IVF due to tubal occlusion. Group 2 included 127 patients who had received laparoscopic ovarian cystectomy for endometrioma(s) followed by IVF treatment. Clinical outcomes of IVF treatment were compared between two groups. RESULTS: More oocytes were harvested per retrieval in Group 1 than Group 2 (p < 0.05). The fertilization rate was higher in Group 1 than Group 2 (p < 0.05). Although the implantation rate was higher in Group 2 (p < 0.05), the clinical pregnancy rate revealed no statistically significant difference between the two groups. CONCLUSIONS: Women who received ovarian cystectomy for endometriomas have fewer oocytes harvested during IVF treatment. However, their chance of pregnancy was comparable to patients with tubal problems who underwent IVF treatment. PMID- 16247713 TI - The significance of antral follicle count in controlled ovarian stimulation and intrauterine insemination. AB - PURPOSE: This retrospective study evaluated the role of antral follicle count (AFC) in predicting ovarian response and successful outcome of stimulated insemination cycles. METHODS: One-hundred and fifty infertile women with bilateral patent tubes receiving a standard regime of human menopausal gonadotrophin (HMG) in their first cycle were evaluated. Multiple regression analysis was used to evaluate the effects of different parameters on ovarian responses and multiple logistic regression analysis was applied to determine which parameters gave the maximum discrimination to predict clinical pregnancy. RESULTS: Body mass index was the only significant parameter to predict the number of follicles > or =14 mm whereas AFC was the only significant parameter to predict the HMG duration. Only the number of follicles > or =14 mm significantly improved the chance of clinical pregnancy with an odds ratio of 1.8. CONCLUSION: AFC was related to the HMG duration but was not predictive of number of follicles and clinical pregnancy of stimulated insemination cycles. PMID- 16247715 TI - Factors affecting assisted reproductive technology (ART) pregnancy rates: a multivariate analysis. AB - PURPOSE: To determine factors instrumental in achieving a clinical pregnancy in assisted reproductive technology (ART) patients. METHODS: This study included 205 women undergoing their first ART cycle. Univariate and multivariate analyses were performed for patient demographics, in vitro production data, and factors associated with embryo transfer. Odds ratios (OR) were performed where appropriate. RESULTS: Our analyses indicated that age (OR: 0.879), specific year in which the cycle was performed (OR: 2.959), and use of intracytoplasmic sperm injection (OR: 2.867) altered potential pregnancy rate. In addition, percent fertilization (OR: 1.028), number of embryos transferred (OR: 1.842), type of catheter used to transfer the embryos (OR: 0.377), presence of blood on the catheter (OR .414), and embryologist (OR: 2.338) also altered pregnancy rate. CONCLUSIONS: Our data indicate patients' age, use of the Cook catheter, and presence of blood on the catheter reduce pregnancy rates. Performing ART in 1999, using ICSI, increasing fertilization rate, increasing number of embryos transferred (albeit less desirable when the chance of multiple gestation may occur), and transferring embryos via a particular embryologist, improve pregnancy rates. PMID- 16247716 TI - Comparison of maturation, meiotic competence, and chromosome aneuploidy of oocytes derived from two protocols for in vitro culture of mouse secondary follicles. AB - PURPOSE: To compare maturation rates of mouse preantral follicles cultured using two previously reported follicle in vitro follicle culture protocols, and to compare the aneuploidy of oocytes derived from the two protocols with in vivo matured control oocytes. METHODS: Mouse preantral follicles were either mechanically isolated then cultured in individual microdroplets, or enzymatically isolated and cultured in groups in a modified culture medium. Maturation of the follicles/oocytes and resulting oocyte aneuploidy rates were evaluated and compared. RESULTS: The mechanical isolation and individual culture protocol (M/I) resulted in higher follicle survival than the enzymatic isolation and group culture protocol (E/G) (89.1% versus 79.1%, p < 0.01), and better maturation to the metaphase 2 stage (61.5% versus 39.5%, p = 0.01) The rate of aneuploidy of oocytes not significantly higher in oocytes from the E/G group than the M/I group (15.4% versus 9.9%), but hypoploidy was significantly increased (4.7% versus 0.9%, p < 0.05). Both groups had a higher rate of aneuploidy than the control oocytes (2.9%, p < 0.02). CONCLUSIONS: These results indicate an increased survival and competency of oocytes derived from the M/I protocol, compared to the E/G protocol. The data highlight an increased susceptibility to meiotic errors in early stage follicles undergoing in vitro culture, compared to in vivo-matured oocytes. PMID- 16247717 TI - Effects of isolating methods (mechanical or enzymatical) on structure of pre antral follicles in mouse. AB - PURPOSE: This study was conducted to evaluate the stable and effective isolating method of pre-antral follicles from ovary by means of mechanical or enzymatical method in mice. METHODS: Mouse (ICR, 3-4 weeks old) ovarian pre-antral follicles of 70-130 microm (A: < or = 90; B: 91-110; C: 111-130 microm) in diameter were isolated by mechanical method (group I) using syringe needles or by enzymatical method (group II) using 600 IU collagenase and DNase and cultured individually in 20 microL droplet of media under mineral oil on culture dish for 8 days. RESULTS: On 8th day after culture, disruption rate (84.4 +/- 18.8% vs. 9.4 +/- 18.8%) was significantly higher (p < 0.05) in group II than in group I. Survival rate (88.0 +/- 28.9% vs. 0%) was significantly higher (p < 0.0000) in group I than in group II. There were no differences in the survival rate among different initial sized pre-antral follicles in group I (A: 86.7 +/- 12.5%, B: 100%, C: 69.2 +/- 30.0%, NS) or group II (A: 0%, B: 0%, C: 0%, NS), respectively. CONCLUSIONS: In conclusion, compared to mechanical dissection, enzymatical isolation of pre antral follicles resulted in higher follicular disruption rate. Survival rate was not affected by different initial sizes of pre-antral follicles in mice. PMID- 16247719 TI - MR imaging of the forefoot: Morton neuroma and differential diagnoses. AB - Magnetic resonance (MR) imaging of Morton neuromas is highly accurate. Morton neuromas are more conspicuous when the patient is prone positioned and the foot is plantar flexed than in the supine position with the toes pointing upward. MR imaging of Morton neuromas has a large influence on the diagnostic thinking and treatment plan of orthopedic foot surgeons. The most common differential diagnoses include intermetatarsal bursitis, stress fractures, and stress reactions. Some diagnoses (nodules associated with rheumatoid arthritis, synovial cyst, soft tissue chondroma, and plantar fibromatosis) are rare and can be diagnosed with histologic correlation only. PMID- 16247718 TI - MTHFR C677T polymorphism associates with unexplained infertile male factors. AB - PURPOSE: To determine whether 5,10-methylenetetrahydrofolate reductase (MTHFR C677T and A1298C) genotype is associated with male infertility. METHODS: Analysis of cytogenetic, Y chromosomal microdeletion assay (Yq), and the C677T and A1298C polymorphisms of the MTHFR gene by pyrosequencing and PCR-Restriction Fragment Length Polymorphism (RFLP) method. SAS 8.1 assessed the statistical risk of MTHFR genotype. RESULTS: The homozygous (T/T) C677T polymorphism of the MTHFR gene was present at a statistically high significance in unexplained infertile men with normal karyotype, instead at no significance in explained infertile men with chromosomal abnormality or Y chromosome deletion. There was no statistically significance of A1298C variation in infertile males. CONCLUSIONS: The MTHFR 677TT genotype may be a genetic risk factor for male infertility, especially with severe OAT and non-obstructive azoospermia in unexplained infertile males. PMID- 16247720 TI - MR imaging of tendons and ligaments of the midfoot. AB - Disorders of ligaments and tendons at the midfoot are less common than those around the ankle and at the hindfoot. Nevertheless, disorders of the distal portions of the anterior and posterior tibialis tendons (ATT and PTT) and the spring ligament complex have an important clinical impact and can be treated with surgery. In this article, we discuss important factors for the magnetic resonance (MR) protocol and review the normal MR anatomy and MR findings in disorders of the distal ATT and PTT as well as of the spring ligament complex. In addition, we discuss clinical aspects of these disorders. PMID- 16247721 TI - MR imaging appearance of rheumatoid arthritis in the foot. AB - Although conventional radiographs remain the initial mainstay for imaging of the foot in patients with rheumatoid arthritis (RA), magnetic resonance (MR) imaging has afforded the ability to detect early signs of the disease (i.e., synovitis, tenosynovitis, bone lesions, and bursitis), especially at the forefoot. In addition, the relatively symmetric distribution of the imaging abnormalities depicted in the metatarsophalangeal joints and the frequent involvement of the retro-calcaneal bursitis are almost specific for RA. In more advanced stages of the disease, MR imaging is well suited to evaluation of the hindfoot joints and tendons as well as the musculoskeletal complications of RA (e.g., tendon disruption, rheumatoid nodules, sinus tarsi syndrome). PMID- 16247722 TI - Stress fractures and related disorders in foot and ankle: plain films, scintigraphy, CT, and MR Imaging. AB - The accurate diagnosis of stress-induced changes in the foot and ankle requires careful and detailed clinical history and examination. This is of paramount importance in ensuring the correct imaging interpretation and for excluding other differential diagnoses. Advanced imaging (scintigraphy and MR imaging) plays a vital role in the early diagnosis of this type of injury, and CT has an important contributory role in the diagnosis of injury where imaging features by the other modalities are equivocal. An early diagnosis reached by judicious use of imaging techniques is the cornerstone of appropriate management, with a high probability of a full recovery, a low risk of complications, and a return to pre-disease activity. PMID- 16247723 TI - Soft tissue masses in the foot and ankle: characteristics on MR Imaging. AB - Benign neoplasms and tumor-like lesions constitute the majority of soft tissue masses in the foot and ankle. Therefore, malignant tumors are often unsuspected at this site and misdiagnosed clinically, especially if occurring in young individuals with unspecific or long-standing clinical symptoms. In addition to radiography, MR imaging represents the method of choice in evaluation of foot tumors. Because of their relatively characteristic imaging appearance, in most cases of benign soft tissue lesions of the foot and ankle a specific diagnosis can be suggested. Unfortunately, malignant tumors can also arise with nonaggressive imaging features. Diagnostic errors can be avoided if any soft tissue lesion that cannot be specifically diagnosed is regarded as potentially malignant until proved otherwise. This article reviews the MR appearance of the most common benign and malignant soft tissue masses in the foot and ankle together with their clinical, radiographic, and pathological findings. PMID- 16247724 TI - Ultrasound of the ankle: anatomy of the tendons, bursae, and ligaments. AB - Ankle tendon and ligament disorders are commonly encountered in everyday orthopedic practice. Whereas tendons can be affected by traumatic, degenerative, and inflammatory conditions, ligaments are mostly involved by tears. Ultrasonography (US) has been accepted worldwide as an efficient, ready, dynamic, and noninvasive tool in assessing ankle tendons and ligaments. The recent technological advances in software and hardware of US equipment have improved the possibilities of US in ankle assessment, and modern equipment allows an accurate depiction of ankle structures. As with all imaging modalities, knowledge of the scanning technique and knowledge of normal US anatomy are prerequisites for a successful examination. In this article we briefly review the normal anatomy of ankle tendons and main ligaments relevant to the US examination. Then we present the routine US technique of examination for the anterior, posterolateral, posteromedial, and posterior region followed by a description of the normal US appearance of tendons, bursae, and ligaments. PMID- 16247725 TI - What the orthopaedic foot and ankle surgeon wants to know from MR Imaging. AB - The study of musculoskeletal disease of the foot and ankle has been revitalized with the advent of cross-sectional imaging modalities. MR imaging provides unsurpassed, high soft tissue contrast resolution and multiplanar capabilities. It offers to the foot and ankle surgeon a quick and noninvasive tool for the diagnosis of a variety of soft tissue and bony disorders that are often difficult to diagnose with alternative modalities. This article focuses on the role of MR imaging in diagnosing ankle and foot disease. PMID- 16247726 TI - Differential diagnosis of pedal osteomyelitis and diabetic neuroarthropathy: MR Imaging. AB - Almost all diabetic foot infections originate from a foot ulcer. Decreased pain perception and structural deformities such as previous partial foot amputation, Charcot joints, and toe deformity in combination with chronic ischemia lead to a propensity for skin breakdown and subsequent infection. Magnetic resonance (MR) imaging is increasingly performed to evaluate for potential bone infection, but diagnosis of osteomyelitis can be complicated because signal changes from acute Charcot arthropathy, fractures, and postoperative residues may be mistaken for infection. Signal alterations of bone infection may be atypical in sclerosing osteomyelitis and gangrene. Differentiation between osteomyelitis and acute or subacute neuroarthropathy requires careful analysis of the location of bone signal alterations, their distribution, and pattern because qualitative changes are often identical. Presence of secondary signs such as adjacent ulcer, cellulitis, and sinus tract is indicative of osteomyelitis. Differentiation of noninfected neuroarthropathy from infected neuroarthropathy based on MR examinations is difficult. Presence of a sinus tract, disappearance of subchondral cysts, diffuse bone marrow abnormality, and bone erosions are in favor of infection. PMID- 16247728 TI - Abbreviations and symbols in peptide science: a revised guide and commentary. AB - The abbreviations and symbols used in Peptide Science are surveyed, with comment and recommendations. PMID- 16247729 TI - Proteome mapping of mature pollen of Arabidopsis thaliana. AB - The male gametophyte of Arabidopsis is a three-celled pollen grain that is thought to contain almost all the mRNAs needed for germination and rapid pollen tube growth. We generated a reference map of the Arabidopsis mature pollen proteome by using multiple protein extraction techniques followed by 2-DE and ESI MS/MS. We identified 135 distinct proteins from a total of 179 protein spots. We found that half of the identified proteins are involved in metabolism (20%), energy generation (17%), or cell structure (12%); these percentages are similar to those determined for the pollen transcriptome and this similarity is consistent with the idea that in addition to the mRNAs, the mature pollen grain contains proteins necessary for germination and rapid pollen tube growth. We identified ten proteins of unknown function, three of which are flower- or pollen specific, and we identified nine proteins whose RNAs were absent from the transcriptome, seven of which are involved in metabolism, energy generation, or cell wall structure. Our work complements and extends recent analyses of the pollen transcriptome. PMID- 16247730 TI - Characterization of microsomal fraction proteome in human lymphoblasts reveals the down-regulation of galectin-1 by interleukin-12. AB - T helper cells (Th) are divided into Th1 and Th2 subsets based upon their cytokine profiles and function. Naive Th cells differentiate into Th1 and Th2 subsets depending on the antigens, costimulatory molecules, and cytokines they encounter. Cytokine interleukin (IL)-12 enhances the generation of Th1 lymphocytes and inhibits the production of Th2 subset. Many genes involved in Th cell differentiation have already been identified at transcriptomic level in microarray studies. In this study, isotope coded affinity tag labeling combined with chromatographic techniques and tandem mass spectrometry was used to find IL 12 regulated proteins in the microsomal fraction of Th cells. A total of 380 and 275 proteins were initially identified and quantitated in two experiments. After the high-confidence protein identifications were restricted to those where at least two different peptides were identified per protein, and these confirmed by manual inspection of the tandem mass spectra, 147 proteins remained. Of these high-confidence protein identifications 41 had at least 1.5-fold change in expression between IL-12 treated and nontreated cells. Among the differentially regulated proteins were galectin-1 (gal-1) and CD7, and their down-regulation was further corroborated with Western blotting and flow cytometry, respectively. Gal 1 and CD7 are known to interact with each other, and regulate immunity through influencing apoptosis and cytokine production. Our data indicate that IL-12 down regulates the expression of both gal-1 and CD7 in the microsomal fraction of peripheral blood mononuclear cells and cord blood CD4(+) cells. The down regulation of these proteins is likely to have a role in specific Th cell selection and cytokine environment creation. PMID- 16247731 TI - The accumulation of abundant soluble proteins changes early in the development of the primary roots of maize (Zea mays L.). AB - A reference database of the major soluble proteins of the primary root of the maize inbred line B73 was generated 5 days after germination (DAG) using a combination of 2-DE and MALDI-TOF MS. A total of 302 protein spots were detected with CBB in a pH 4-7 range and 81 proteins representing 74 distinct Genbank accessions were identified. Only 28% of the major proteins identified in 5 DAG primary roots were identified in similarly analyzed 9 DAG primary roots documenting remarkable changes in the accumulation of abundant soluble proteins early in primary root development. PMID- 16247732 TI - Characterization of pollen tube development in Pinus strobus (Eastern white pine) through proteomic analysis of differentially expressed proteins. AB - The differentially expressed proteins in pollen tubes indicate their specific roles in this stage of male gametophyte development. To isolate these proteins, 2 DE was done using ungerminated pollen and 2-day-old pollen tubes of Pinus strobus. Results show that 645 and 647 protein spots were clearly resolved from pollen grains and pollen tubes, respectively. Thirty-eight protein spots were expressed only in pollen tubes, while 19 increased in intensity. MALDI-TOF MS was used to generate tryptic peptide masses that were submitted to Mascot for identification. Of the differentially expressed proteins, 12% matched with hypothetical proteins, 33% did not hit any protein, and for the 55%, a putative function was assigned based on similarity of sequences with previously characterized proteins. Therefore, pollen tube development can be characterized by the cellular activities that involve metabolism, stress/defense response, gene regulation, signal transduction, and cell wall formation. This study expands our understanding of the changes in protein expression associated with pollen tube development and provides insights into the molecular programs that separate the development of the pollen tubes from pollen grains. This is the first report that describes a global analysis of differentially expressed proteins from the pollen tube of any seed plant. PMID- 16247733 TI - Proteomic analysis of enriched microsomal fractions from GS-NS0 murine myeloma cells with varying secreted recombinant monoclonal antibody productivities. AB - The folding, transport and modification of recombinant proteins in the constitutive secretory pathway of eukaryotic cell expression systems are reported to be a bottleneck in their production. We have utilised a proteomic approach to investigate the processes catalysed by proteins constituting the secretory pathway to further our understanding of those processes involved in high-level antibody secretion. We used GS-NS0 cell populations differing in qmAb to prepare enriched microsome fractions from each cell population at mid-exponential growth phase. These were analysed by 2-D PAGE to characterise the microsome protein component and test the hypothesis that bottlenecks in recombinant protein synthesis exist in these compartments, which are alleviated in high producers by the up-regulation of key secretory pathway proteins. Proteins whose abundance changed in a statistically significant manner with increasing qmAb were involved in a range of cellular functions: energy metabolism, mAb folding/assembly, cytoskeletal organisation and protein turnover. Amongst these were BiP and PDI, chaperones resident in the ER that interact with nascent immunoglobulins during their folding/assembly. However, our results suggest that there are diverse mechanisms by which these cells achieve qmAb. The results imply that cell engineering strategies for improving qmAb should target proteins associated with altered functional phenotype identified in this study. PMID- 16247734 TI - Proteomic analysis of organ-specific post-translational lysine-acetylation and methylation in mice by use of anti-acetyllysine and -methyllysine mouse monoclonal antibodies. AB - Post-translational lysine-acetylation and -methylation are two major PTMs of lysine residues in proteins. Recently, we established pan-reactive anti acetyllysine mouse mAbs, which can bind to Nepsilon-acetylated lysine residues in various contexts of amino acid sequences. In the present study, we established pan-reactive anti-methyllysine mouse mAbs comparable to the anti-acetyllysine ones. By using these anti-acetyllysine and -methyllysine antibodies, we found that the pattern of lysine-acetylated and -methylated proteins in mouse organs showed extreme variation from organ to organ. We selected brain and skeletal muscle as model cases to be further analyzed by 2-DE followed by Western blotting. In brain, alpha-tubulin at its basal level was found to be extremely acetylated; and alpha-enolase was shown to be a newly recognized possibly acetylated protein. NF-L protein, Hsc70, alpha-tubulin fragments, beta-actin, and brain-type creatine kinase were identified as putative lysine-methylated proteins in mouse brain. In skeletal muscle, lysine-methylation of alpha-actin and both lysine-acetylation and -methylation of muscle-type creatine kinase were found as novel putative lysine-modified proteins. The approach presented here might be useful to find novel disease markers and/or drug target molecules that would not be noticed by use of the traditional proteomic approach only. PMID- 16247735 TI - Proteomic analysis using an unfinished bacterial genome: the effects of subminimum inhibitory concentrations of antibiotics on Mannheimia haemolytica virulence factor expression. AB - Here we identify, using nonelectrophoretic proteomics, effects of subminimum inhibitory concentrations (subMIC) of two antibiotic preparations, chlortetracycline (CTC), and chlortetracycline-sulfamethazine (CTC + SMZ), on protein expression in the bovine respiratory pathogen Mannheimia haemolytica. The M. haemolytica genome is currently in draft form, and annotation is incomplete. Relying on the principle of gene sequence conservation across species, we used annotated genomes from closely related species to identify, confirm, and functionally annotate 495 M. haemolytica proteins. To conduct quantitative comparative proteomics, we developed a protein quantitation method based on the cross correlation function of the SEQUEST algorithm. When M. haemolytica was cultivated in the presence of 1/4 MIC of CTC and CTC + SMZ, expression of proteins involved in energy production, nucleotide metabolism, translation, and the bacterial stress response (chaperones) were affected. The most notable subMIC effect was a significant decrease in the expression of leukotoxin A, which is an important M. haemolytica virulence factor. Reduction in leukotoxin expression could be one of the molecular mechanisms responsible for the efficacy of these antibiotics against bovine respiratory disease. PMID- 16247736 TI - Prenatal observation of fetal defecation using four-dimensional ultrasonography. PMID- 16247737 TI - Application of glycine in acute alcohol hallucinosis. AB - INTRODUCTION: Previous studies showed that patients with acute alcohol hallucinosis had significantly lower levels of the inhibitory amino acid neurotransmitter, glycine. METHODS: A placebo-controlled, double-blind study of glycine in 40 patients with acute alcohol hallucinosis. Treatment duration was 7 days with no other psychotropic medication. RESULTS: The 20 patients randomised to the active drug (700 mg glycine sublingually) demonstrated a significant decrease in severity of hallucinosis compared to the 20 patients receiving placebo. CONCLUSION: The positive effects of glycine may be related to an altered balance between excitatory and inhibitory amino acid neurotransmitters. PMID- 16247738 TI - Mapping complex tissue architecture with diffusion spectrum magnetic resonance imaging. AB - Methods are presented to map complex fiber architectures in tissues by imaging the 3D spectra of tissue water diffusion with MR. First, theoretical considerations show why and under what conditions diffusion contrast is positive. Using this result, spin displacement spectra that are conventionally phase encoded can be accurately reconstructed by a Fourier transform of the measured signal's modulus. Second, studies of in vitro and in vivo samples demonstrate correspondence between the orientational maxima of the diffusion spectrum and those of the fiber orientation density at each location. In specimens with complex muscular tissue, such as the tongue, diffusion spectrum images show characteristic local heterogeneities of fiber architectures, including angular dispersion and intersection. Cerebral diffusion spectra acquired in normal human subjects resolve known white matter tracts and tract intersections. Finally, the relation between the presented model-free imaging technique and other available diffusion MRI schemes is discussed. PMID- 16247739 TI - A unified magnetic resonance imaging pharmacokinetic theory: intravascular and extracellular contrast reagents. AB - A fundamental reworking of pharmacokinetic theory for the use of contrast reagents (CRs) in T(1)-weighted MRI studies is presented. Unlike the standard model in common use, this derivation starts with the quantities measured, the intravascular, interstitial, and intracellular (1)H(2)O signals. The time dependences of CR concentrations are introduced as perturbations of the T(1) values of these. Since there is an explicit accounting for the equilibrium exchange of water molecules between tissue compartments, the approach here is a new (second) generation of the shutter-speed model (S(2)M). When the first-order rate constant measuring CR extravasation (K(trans)) is of sufficient magnitude, simulations presented here confirm that neglect of plasma CR, a feature of the first generation of S(2)M, is a valid approximation. The second S(2)M generation (S(2)M2) also automatically accommodates excursions of either or both of the two major equilibrium water exchange systems (transendothelial and transcytolemmal) into any or all possible exchange conditions, from their fast-exchange limits to their slow-exchange limits. This can happen not because the exchange kinetics themselves vary during the isothermal CR passage, but because the MR shutter speeds for these processes can vary. When K(trans) is sufficiently small, the S(2)M2 also naturally accounts for the hyperfine blood agent level dependent (BALD) effect that is easily detectable at high magnetic field. This can be seen for virtually all CRs in normal brain tissue and for virtually all tissues with sufficiently intravascular CRs. Thus, S(2)M2 represents a unified pharmacokinetic theory for intravascular and extracellular T(1) contrast reagents. PMID- 16247740 TI - von Willebrand disease R1374C: type 2A or 2M? A challenge to the revised classification. High frequency in the northwest of Spain (Galicia). AB - Patients initially diagnosed with type 1 von Willebrand disease (VWD) have been reclassified as type 2 after a more exhaustive analysis in several studies. Our study's objectives were (1) to reanalyze patients that were previously diagnosed as type 1 to achieve a more accurate diagnosis and (2) to compare the von Willebrand factor (VWF) ristocetin cofactor assay (VWF:RCo) and the VWF collagen binding assay (VWF:CB) in order to evaluate the possibility of replacing the former assay with the latter in the diagnosis of VWD. Twenty-one patients from two large unrelated families and 104 normal controls were studied. VWF:Ag, VWF:RCo, FVIII coagulant activity (FVIII:C), bleeding time (BT), PFA(100), and multimeric analysis of VWF were tested. Genetic analysis by sequencing exon 28 on the VWF gene was also carried out. Patients presented lower levels of VWF:Ag and VWF:RCo, a dissociation between VWF:RCo/VWF:Ag, and the presence of all sizes of multimers in plasma VWF. The results for VWF:CB varied depending on the type of collagen used. The genetic analysis showed that the mutation R1374C is responsible for type 2M VWD. A high frequency of the R1374C mutation is observed in northwestern Spain (Galicia). Some types of 2M VWD are misdiagnosed as type 1 VWD. The VWF:CB (with type I collagen) assay was unable to discriminate defective platelet binding of the R1374C VWF. This confirms that VWF:CB cannot substitute for VWF:RCo, and both should be tested when diagnosing VWD. PMID- 16247741 TI - Intravenous gamma globulin is effective as an urgent treatment in Brucella induced severe thrombocytopenic purpura. AB - Severe thrombocytopenia is a rare hematologic manifestation of brucellosis, which can occasionally be associated with bleeding into the skin and from mucosal sites. Prompt recognition of this brucellosis complication and aggressive therapy is vital because the mortality rate associated with bleeding into the central nervous system is high. We report a case of a patient infected with Brucella melitensis who was admitted with a severe case of thrombocytopenic purpura. The patient responded well to intravenous gamma globulin (IVIg) treatment with platelet recovery within 2-3 days. For cases of Brucella-induced thrombocytopenic purpura, IVIg may be administered as an urgent therapy until the microbial therapy takes effect. PMID- 16247742 TI - Successful umbilical cord blood transplantation for severe chronic active Epstein Barr virus infection after the double failure of hematopoietic stem cell transplantation. AB - An 11-year-old boy with severe chronic active Epstein-Barr virus infection (CAEBV) underwent successful cord blood transplantation (CBT) after consecutive failure of peripheral blood and bone marrow transplantation from his HLA mismatched mother. CB cells from an unrelated donor were infused after conditioning with total body irradiation (12 Gy), melphalan (120 mg/m(2)), and etoposide (600 mg/m(2)). Complete remission without circulating EBV-DNA has continued for 15 months after a delayed hematologic recovery. This is the first successful report of CBT for CAEBV. CB may therefore be an alternate source of stem cells for the curative treatment of CAEBV, despite the absence of EBV specific cytotoxic T lymphocytes. PMID- 16247743 TI - Guidance on the use of miglustat for treating patients with type 1 Gaucher disease. AB - Type 1 Gaucher disease (GD) is a progressive lysosomal storage disorder due to an autosomal recessive deficiency of glucocerebrosidase. Clinical manifestations include anemia, thrombocytopenia, hepatosplenomegaly, and bone and pulmonary disease. Intravenous enzyme replacement (ERT) with imiglucerase is the accepted standard for treatment of symptomatic patients. More than 3,500 patients worldwide have received ERT with well-documented beneficial effects on the hematological, visceral, skeletal, and pulmonary manifestations, and with resultant improvement in health-related quality of life. Miglustat, an imino sugar that reversibly inhibits glucosylceramide synthase and reduces intracellular substrate burden, is an oral treatment for patients with type 1 GD that was recently approved in the United States for symptomatic patients with mild to moderate clinical manifestations for whom ERT is not an option. Because responses to miglustat are slower and less robust than those observed with ERT, and because miglustat is associated with significant side effects, clinicians who care for patients with GD should become familiar with the limited indications for miglustat use and the circumstances when it may be prescribed appropriately. This review article and position statement represents the current opinion of American physicians with extensive expertise in GD regarding patient management in the context of the availability of standard imiglucerase treatment and the recent introduction of miglustat. PMID- 16247744 TI - Primary central nervous system lymphoma following liver transplantation. PMID- 16247745 TI - High-dose cyclophosphamide and MESNA infusion can cause acute atrial fibrillation. PMID- 16247746 TI - Prevention of hepatitis B reactivation with lamivudine in hepatitis B virus carriers with hematologic malignancies treated with chemotherapy--a prospective case series. AB - Administration of immunosuppressive treatment in hepatitis B virus carriers with malignancies is associated with the risk of hepatitis B reactivation. This complication is more frequent in patients with hematologic malignancies because administration of corticosteroids, the mainstay of treatment of these patients, is an independent risk factor for hepatitis B reactivation. When lamivudine is given prior to chemotherapy, it prevents the viral replication during the immunosuppression period; therefore, it might reduce the risk of hepatitis B exacerbation. We performed a prospective study to assess the efficacy of prophylactic administration of lamivudine in this setting. Ten hepatitis B virus carriers with hematologic malignancies were included in this study; seven were HBsAg positive, and three had isolated antiHBc and detectable HBV-DNA levels. Nine patients were given corticosteroids after the administration of lamivudine. Lamivudine was given per os at a dose of 100 mg once daily. In four patients that had not been previously treated with chemotherapy, lamivudine was started 19 days (median) (range, 0-35 days) prior to the onset of chemotherapy. The administration of lamivudine has not stopped since in any of our patients. After a median follow-up of 15 months (range 6-38 months), no hepatitis B reactivation was observed. HBV-DNA levels were decreased in all 6 patients who had detectable HBV-DNA at baseline. Lamivudine was well tolerated. Chemotherapy regimens were administered as planned, and their effectiveness was not compromised by lamivudine. In conclusion, prophylactic administration of lamivudine should be considered as a means of reducing the frequency of hepatitis B reactivation in hepatitis B virus carriers with hematologic malignancies who are being treated with chemotherapy. PMID- 16247747 TI - T-Cell lymphoblastic lymphoma presenting as bilateral multinodular breast masses: a case report and review of the literature. AB - Non-Hodgkin lymphoma of T-cell lineage involving the breast is rare. We report on a 41-year-old woman with T-cell lymphoblastic lymphoma who presented with multiple bilateral breast masses. The patient was treated with intensive chemotherapy and mediastinal and whole-brain irradiation. She remains in complete remission 24 months after diagnosis. The clinical, histologic, phenotypic, and cytogenetic features are described, with a review of the literature. PMID- 16247748 TI - Thrombocytopenic conditions-autoimmunity and hypercoagulability: commonalities and differences in ITP, TTP, HIT, and APS. AB - Immune thrombocytopenia purpura (ITP), thrombotic thrombocytopenia purpura (TTP), heparin-induced thrombocytopenia (HIT), and antiphospholipid syndrome (APS) are clinical conditions associated with significant morbidity and mortality. These well-defined clinical syndromes have in common several properties: (1) their pathogenesis is immune mediated, specifically by autoantibodies; (2) thrombocytopenia is a hallmark in these four conditions; (3) except for the case of ITP, platelet and endothelial cell activation occurs in TTP, HIT, and APS, resulting in a prothrombotic state and an increased risk of thrombosis. Although these four immune-mediated syndromes are well-defined diseases, several case reports and studies have documented the association of two diseases in the same patient, illustrating the concept of the kaleidoscope of autoimmunity. PMID- 16247749 TI - Thrombotic thrombocytopenic purpura induced by cyclosporin a after allogeneic bone marrow transplantation treated by red blood cell exchange transfusion: a case report. PMID- 16247750 TI - Overexpression of cyclooxygenase-2 in multiple myeloma: association with reduced survival. AB - Cyclooxygenases (COX) are key enzymes in the conversion of arachidonic acid to prostaglandins. Several studies have shown a relation between angiogenesis and COX-2 expression. Elevated expression of cyclooxygenase-2 (COX-2), however, has not been reported in multiple myeloma (MM) in the literature. The aim of this study is to investigate COX-2 expression in MM as well as its correlation with prognostic factors and estimated survival rates. Immunohistochemical staining of the paraffin-embedded bone marrow biopsy tissues (n = 51) was performed using isoform-specific COX-2 polyclonal antisera (Santa Cruz Biotechnology, Santa Cruz, CA). Results were correlated with recognized clinical parameters, which were retrospectively obtained from patients' files. There were 15, 19, and 17 bone marrow biopsy specimens with negative, weak-moderate, and strong COX-2 immunostaining, respectively. According to univariate analysis, beta2 microglobulin, age, stage, COX-2 expression, and serum lactate dehydrogenase levels were significant prognostic factors for survival in patients with multiple myeloma. COX-2 expression, age, and serum lactate dehydrogenase levels (greater than 1x normal level) were significant prognostic factors by multivariate analysis. Kaplan-Meier overall survival estimate of those patients with negative or weak-moderate COX-2 immunoreactivity in myeloma cells was significantly better than that of patients with strong COX-2 immunoreactivity (log-rank chi(2) = 21,43, P < 0.001). COX-2 overexpression was associated with reduced estimated survival. Poor prognostic factors such as LDH, age, and beta2-microglobulin were also correlated with COX-2 expression. Potent, specific COX-2 inhibitors showing evident antiangiogenic and antitumor effects on cancers could provide new therapeutic strategies in the treatment of MM. PMID- 16247751 TI - Relationship of B-type natriuretic peptide and anemia in patients with and without heart failure: a substudy from the Breathing Not Properly (BNP) Multinational Study. AB - While anemia is a significant risk factor for poor outcomes in patients with heart failure (HF), it is not in defined guidelines for HF assessment. B-type natriuretic peptide (BNP) is a marker for diagnosis and management of patients with HF. We determined the incidence of anemia in patients with HF and the relationship between BNP and hemoglobin (Hgb) levels in patients with and without HF. Results from the Breathing Not Properly Multinational Trial consisted of 1,586 patients presenting to the emergency department (ED) with dyspnea. Because renal insufficiency is a confounding variable for BNP, patients with a creatinine of >or=2.0 mg/dL were excluded. The remaining data were evaluated from 620 non-HF patients (337 M, 283 F) and 547 HF patients (299 M, 248 F). The New York Heart Association (NYHA) HF classification and ejection fraction by echocardiography were assessed for HF patients. Blood was tested for Hgb, BNP, and creatinine. Using World Health Organization criteria for anemia, we observed that HF patients in NYHA class III or IV had lower mean Hgb levels (12.5 g/dL, P < 0.05) and a higher incidence of anemia (48.2%, P < 0.05) than did HF patients in class I or II (13.4 g/dL and 33.9%, respectively). There was no correlation between Hgb and log BNP for females without HF or the aggregate of all HF patients. In contrast, a significant inverse correlation was observed for males without HF (P < 0.001). Although there were differences in the BMI, age, and estimated glomerular filtration rate (eGFR) versus Hgb observed in this group, the log BNP correlation remained significant after multivariate analysis. A significant inverse correlation for log BNP and Hgb were also observed for diastolic (EF >or= 50) HF (P < 0.05) that was also not accounted for by the BMI, age, or eGFR. The presence of anemia is associated with worsening HF at ED presentation. For males without HF and diastolic HF patients of both genders, a low Hgb may be a confounding variable toward increasing BNP. Among systolic HF patients, the presence of a low hemoglobin concentration is not a factor in the interpretation of BNP results. PMID- 16247752 TI - Minimal residual disease (MRD) monitoring using rearrangement of T-cell receptor and immunoglobulin H gene in the treatment of adult acute lymphoblastic leukemia patients. AB - We evaluate whether molecular monitoring of minimal residual disease (MRD) using TCR delta (TCRD), TCR gamma (TCRG), and immunoglobulin H (IgH) gene rearrangements in the bone marrow (BM) is correlated with clinical events in ALL patients. The 14 patients enrolled in this study included 6 males and 8 females with a median age of 53 years (range, 25-79 years), and the median duration of follow-up was 417 days (range, 57-617 days). The median WBC count was 11.3 x 10(9)/L at diagnosis. All patients had L2 type ALL. Eleven patients had a monoclonal pattern of IgH (7), TCRD (3) and TCRG (1), and 3 patients had two clonal patterns. Eleven of the 14 patients achieved the first complete remission (CR) after the first induction chemotherapy. We analyzed 9 of 11 CR patients who could be examined immediately after induction chemotherapy (including re induction therapy). Event-free survival (EFS, 0%) and disease-free survival (DFS, 0%) at 1 year in CR patients with MRD level >or=10(-3) (n = 3) were significantly lower than those in CR patients with MRD level <10(-3) (n = 6) (log-rank test, P = 0.013, 0.013). A lower MRD in BM value after induction chemotherapy was associated significantly with longer survival in the log-rank test. Our data provide evidence that molecular MRD status of BM is a strong predictor of outcome in adult ALL. PMID- 16247754 TI - Intense paraneoplastic neutrophilic leukemoid reaction related to a G-CSF secreting lung sarcoma. AB - A white blood cell count more than 50 x 10(9)/l, not related to bone marrow involvement, is termed leukemoid reaction. We report on the first case of an undifferentiated sarcoma of the lung associated with an intense paraneoplastic neutrophilic leukemoid reaction related to the production of granulocyte colony stimulating factor (G-CSF). A radiography and a computed tomography scan of the chest revealed a well-limited voluminous and heterogeneous low-density mass of the left lung. The patient died of multiorgan failure related to uncontrolled progressive tumor growth after admission and two cycles of chemotherapy. The patient's G-CSF serum concentration was dramatically elevated (6,538 pg/ml) compared to serum levels observed in normal controls and patients with elevated leukocytosis (31 and 387 pg/ml, respectively). The G-CSF concentration dramatically increased after the first cycle of chemotherapy and during the subsequent neutropenia, as a result of the tumor lyses as well as of disruption of the physiological negative feedback mechanism. Adjunction of the patient's serum to CD34+ cell cultures induced a 12.3-fold increase in CD15+ cells, demonstrating the serum's capacity to induce myeloid differentiation. PMID- 16247755 TI - Possible association between Budd-Chiari Syndrome and gemtuzumab ozogamicin treatment in a patient with refractory acute myelogenous leukemia. AB - Gemtuzumab ozogamicin (GO; CMA-676; Mylotarg) is a chemotherapeutic agent approved for the treatment of CD33-positive acute myelogenous leukemia in patients of age 60 years or older after first relapse. Hepatic veno-occlusive disease has been reported to develop as a late complication of gemtuzumab ozogamicin treatment. A patient who developed Budd-Chiari Syndrome with hepatic vein thrombosis following treatment with GO is presented. This complication has not been previously reported, and it deserves to be considered as a possible adverse effect of gemtuzumab ozogamicin. PMID- 16247756 TI - Prenatal detection of associated anomalies in fetuses diagnosed with cleft lip with or without cleft palate in utero. AB - OBJECTIVE: The aim of this study was to determine the prenatal detection rate of associated anomalies in fetuses with a suspected cleft lip with or without cleft palate. METHODS: Fetuses with a suspected cleft lip with or without cleft palate, determined by prenatal ultrasound, were prospectively enrolled. Additional anomalies suspected by ultrasound or genetic testing were recorded. Postnatal outcome was obtained. RESULTS: Forty-five fetuses with a cleft lip with or without cleft palate, diagnosed prenatally with either two-dimensional and/or three-dimensional ultrasound, were studied. Postnatal follow-up revealed that 16 (35.6%) of these 45 fetuses had an additional structural or syndromic abnormality. Of the 37 fetuses with prenatally determined 'isolated' cleft lip with or without cleft palate, eight (21.6%) had an additional malformation identified after delivery. CONCLUSION: In pregnancies complicated by a cleft lip with or without cleft palate, patients should be informed of the risks of associated anomalies, some of which may be undetected prenatally. PMID- 16247757 TI - Thymopoietin (lamina-associated polypeptide 2) gene mutation associated with dilated cardiomyopathy. AB - Thymopoietin or TMPO (indicated by its alternative gene symbol, LAP2, in this work) has been proposed as a candidate disease gene for dilated cardiomyopathy (DCM), since a LAP2 product associates with nucleoplasmic lamins A/C, which are encoded by the DCM gene LMNA. We developed a study to screen for genetic mutations in LAP2 in a large collection of DCM patients and families. A total of 113 subjects from 88 families (56 with familial DCM (FDC) and 32 with sporadic DCM) were screened for LAP2 mutations using denaturing high-performance liquid chromatography and sequence analysis. We found a single putative mutation affecting the LAP2alpha isoform in one FDC pedigree. The mutation predicts an Arg690Cys substitution (c.2068C>T; p.R690C) located in the C-terminal domain of the LAP2alpha protein, a region that is known to interact with lamin A/C. RT-PCR, Western blot analyses, and immunolocalization revealed low-level LAP2alpha expression in adult cardiac muscle, and localization to a subset of nuclei. Mutated Arg690Cys LAP2alpha expressed in HeLa cells localized to the nucleoplasm like wild-type LAP2alpha, with no effect on peripheral and nucleoplasmic lamin A distribution. However, the in vitro interaction of mutated LAP2alpha with the pre lamin A C-terminus was significantly compromised compared to the wild-type protein. LAP2 mutations may represent a rare cause of DCM. The Arg690Cys mutation altered the observed LAP2alpha interaction with A-type lamins. Our finding implicates a novel nuclear lamina-associated protein in the pathogenesis of genetic forms of dilated cardiomyopathy. PMID- 16247759 TI - Direct Fmoc/tert-Bu solid phase synthesis of octamannosyl polylysine dendrimer peptide conjugates. AB - The mannose binding proteins on the surface of the dendritic cells are responsible for capture of pathogens in the early stages of immune response. Conjugation to mannose dendrimers is a rarely explored but potentially powerful strategy for enhancing immunogenicity of synthetic peptides relying on direct delivery to dendritic cells. We describe a general protocol for preparation of pure, monodisperse third-generation mannosylated poly-L-lysine dendrimer-peptide conjugates using direct, machine-assisted Fmoc/t-Bu solid phase peptide synthesis. The glycodendrons were elaborated onto the N- or C-terminus of sequences derived from HIV-1 gp41, SARS-CoV S2 protein, and Influenza Hemagglutinin (consisting of 15-44 residues). The products were obtained in a homogeneous state after cleavage from the resin, deprotection, and a single purification on semipreparative RP-HPLC. PMID- 16247760 TI - Regional differences of the prefrontal cortex in pediatric PTSD: an MRI study. AB - Previous studies have revealed altered structural development of the frontal lobes and prefrontal cortex (PFC) in children with symptoms of posttraumatic stress disorder (PTSD). This study is the first to provide a detailed structural analysis of the PFC in children with and without PTSD symptoms. We compared gray and white matter volume in four subregions of the PFC between said groups, then explored whether volume was associated with PTSD symptom severity and functional impairment. PFC measurements were extracted from magnetic resonance imaging (MRI) data from a sample of 23 children (ages 7-14) with a history of trauma and symptoms of PTSD, who had undergone assessment for PTSD symptoms and functional impairment using the Child and Adolescent version of the Clinician-Administered PTSD Scale (CAPS-CA). These measurements were compared to data from an age equivalent control group of 24 healthy children. Children with PTSD symptoms showed a significantly larger volume of gray matter in the delineated middle inferior and ventral regions of the PFC than did control children. Decreased volume of gray matter in the dorsal PFC correlated with increased functional impairment scores. Results indicate that increased volume of the middle-inferior and ventral PFC may be associated with trauma and PTSD symptoms in children. Furthermore, the neuroanatomy of the dorsal PFC may influence the degree of functional impairment experienced by children with PTSD symptoms. PMID- 16247761 TI - Toward a rational design of beta-peptide structures. AB - Intrinsic conformational characteristics of beta-peptides built up from simple achiral and chiral beta-amino acid residues (i.e., HCO-beta-Ala-NH2, HCO-beta-Abu NH2) were studied using quantum chemical calculations and 1H-NMR spectroscopy. A conformer-based systematic and uniform nomenclature was introduced to differentiate conformers. Geometry optimizations were performed on all homoconformers of both HCO-(beta-Ala)(k)-NH2 and HCO-(beta-Abu)(k)-NH2 (1 < or = k < or = 6) model systems at the RHF/3-21G and RHF/6-311++G(d, p) levels of theory. To test for accuracy and precision, additional computations were carried out at several levels of theory [e.g., RHF/6-31G(d), and B3LYP/6-311++G(d, p)]. To display the folding preference, the relative stability of selected conformers as function of the length of the polypeptide chain was determined. Ab initio population distribution of hexapeptides and the conformational ensemble of synthetic models composed of beta-Ala and beta-Abu studied using 1H-NMR in different solvents were compared at a range of temperatures. Helical preference induced by various steric effects of nonpolar side chains was tested using higher level ab initio methods for well-known model systems such as: HCO-(beta-HVal-beta HAla-beta-HLeu)2-NH2, HCO-(ACHC)6-NH2, HCO-(trans-ACPC)6-NH2, and HCO-(cis-ACPC)6 NH2. The relative stabilities determined by theoretical methods agreed well with most experimental data, supporting the theory that the local conformational preference influenced by steric effects is a key determining factor of the global fold both in solution and in the gas phase. PMID- 16247762 TI - Multicolor spectral karyotyping of the L5178Y Tk+/- -3.7.2C mouse lymphoma cell line. AB - The L5178Y/Tk+/- -3.7.2C mouse lymphoma cell line is characterized, at the cytogenetic level, by a karyotype involving both numerical and complex structural aberrations. While the karyotype is remarkably normal for a transformed cell line that has been in culture for almost half a century, there are a number of chromosomal alterations that because of their complexity cannot be fully characterized by routine or even high-resolution G-banding studies. Multicolor spectral karyotyping (SKY) was performed on the cell line in anticipation of identifying the previously unresolved chromosome aberrations and confirming interpretations previously identified by banding studies. New chromosome aberrations detected by SKY include numerical aberrations of chromosome 15, duplications of regions of chromosomes 4, 5, 12, and 18, and deletion of chromosome 14. Complex unbalanced translocations involved segments of chromosomes 6, 14, and 15. In total, the SKY technique was able to provide new refined designations on segments of eight different chromosome pairs (4, 5, 6, 9, 12, 14, 15, 18) and identified all three previously unidentified marker chromosomes. This analysis provides an updated standard reference for the karyotype of the L5178Y/Tk+/- -3.7.2C cell line used in the in vitro mouse lymphoma mutation assay. PMID- 16247763 TI - Mutation spectrum in UVB-exposed skin epidermis of a mildly-affected Xpg deficient mouse. AB - A C-terminal 183 amino acid-truncated mutation of the mouse Xpg gene (XpgDeltaex15) gives rise to a partial deficiency in nucleotide excision repair in homozygously affected cells. We studied the effect of this mutation on UVB induced mutagenesis in mouse skin, using transgenic mice harboring lambda-phage based bacterial lacZ genes as a mutational reporter. UVB increased the lacZ mutant frequency in the epidermis moderately in the homozygous mutant mice, but significantly higher than in the wild-type or the heterozygous mice, whereas background mutant frequencies were not appreciably different among the three mouse genotypes. Ninety-eight lacZ mutant sequences isolated from the UVB-exposed epidermis of the XpgDeltaex15-homozygous mice were analyzed and compared with mutant sequences from the wild-type mice. The spectra of the mutations in the two mouse genotypes were not significantly different, and they were highly UV specific. There were frequent C --> T transitions at dipyrimidine sites and several CC --> TT tandem mutations, although the UV-specific mutations occurred more frequently at CpG sites in the mutant mice. The distribution of the mutations observed in the lacZ transgene and the preferred sequence context of the UV-specific C --> T mutations (5'-TC-3' > 5'-CC-3' > 5'-CT-3') in the Xpg mutant mice were similar to those found in the wild-type mice. Despite these similarities, we detected a previously unrecognized type of the UV-induced mutation only in the Xpg mutant (6/98 in the mutation spectrum of the mutant vs. 0/76 in the wild-type; P = 0.035), which is characterized by multiple base substitutions or frameshifts within a three-nucleotide sequence containing a dipyrimidine. We propose that this putative new class of mutation, which we refer to as a "triplet mutation", is characteristic of UV-induced mutation in an excision-repair-deficient background. PMID- 16247764 TI - An investigation into the effect of the type IV phosphodiesterase inhibitor rolipram in the modulation of glutamate release from rat prefrontocortical nerve terminals. AB - The present study was conducted to explore the influence of rolipram, a specific inhibitor of the phosphodiesterase type 4 (PDE4) isoform, on glutamate release in the rat prefrontal cortex, using isolated nerve terminal (synaptosome) preparation. In prefrontocortical nerve terminals, rolipram potentiated the Ca(2+)-dependent release of glutamate evoked by 4-aminopyridine (4AP) in a concentration-dependent manner. This potentiation of release was occluded by the activation of PKA by Sp-cAMP or beta-adrenergic receptor agonist and prevented by the inhibition of PKA by Rp-cAMP or KT5720, indicating a PKA-mediated mechanism. The rolipram-mediated potentiation of glutamate release is associated with an increase both in the 4AP-evoked depolarization of the synaptosomal plasma membrane potential and in 4AP-evoked Ca(2+) influx into synaptosomes. Moreover, Ca(2+) ionophore ionomycin-induced glutamate release was also facilitated by rolipram. These results concluded that phosphodiesterase 4 inhibited by rolipram produces an increase in PKA activation, which subsequently enhances the voltage dependent Ca(2+) influx by increasing terminal excitability as well as the vesicular release machinery to cause an increase in evoked glutamate release from rat prefrontocortical nerve terminals. PMID- 16247765 TI - Increased expression of calcium/calmodulin-dependent protein kinase IIbeta in frontal cortex in schizophrenia and depression. AB - In searching for genes dysregulated in schizophrenia, we measured the expression of the two splice variants of calcium/calmodulin-dependent protein kinase II (CaMKIIalpha and CaMKIIbeta) in postmortem frontal cerebral cortex tissues from patients who had died with schizophrenia, bipolar disorder, or severe depression. The mRNA levels of expression of these two splice variants were measured by real time Quantitative PCR, using an Mx4000 instrument. The values for the expression of CaMKIIalpha and CaMKIIbeta were normalized by the expression of beta glucuronidase in the tissues. The expression of CaMKIIalpha was significantly elevated in the depression tissues by 29%. The expression of CaMKIIbeta was significantly elevated in the schizophrenia tissues by 27%, and in the depression tissues by 36%. Because CaMKIIbeta influences the expression of many neuroreceptors and influences neural outgrowth and pruning, its altered expression in the cerebral cortex in schizophrenia or depression may contribute to these diseases. PMID- 16247767 TI - Nitric oxide modulates neuromuscular transmission during hypoxia in rat diaphragm. AB - Hypoxia impairs neuromuscular transmission in the rat diaphragm. In previous studies, we have shown that nitric oxide (NO) plays a role in force modulation of the diaphragm under hypoxic conditions. The role of NO, a neurotransmitter, on neurotransmission in skeletal muscle under hypoxic conditions is unknown. The effects of the NO synthase (NOS) inhibitor nomega-nitro-L-arginine (L-NNA, 1 mM) and the NO donor spermine NONOate (Sp-NO, 1 mM) were evaluated on neurotransmission failure during nonfatiguing and fatiguing contractions of the rat diaphragm under hypoxic (PO2 approximately 5.8 kPa) and hyperoxic conditions (PO2 approximately 64.0 kPa). Hypoxia impaired force generated by both muscle stimulation at 40 HZ (P40M) and by nerve stimulation at 40 HZ (P40N). The effect of hypoxia in the latter was more pronounced. L-NNA increased P40N whereas Sp-NO decreased P40N during hypoxia. In contrast, neither L-NNA nor Sp-NO affected P40N during hyperoxia. L-NNA only slightly reduced neurotransmission failure during fatiguing contractions under hyperoxic conditions. Consequently, neurotransmission failure assessed by comparing force loss during repetitive nerve simulation and superimposed direct muscle stimulation was more pronounced in hypoxia, which was alleviated by L-NNA and aggravated by Sp-NO. These data provide insight in the underlying mechanisms of hypoxia-induced neurotransmission failure. This is important as respiratory muscle failure may result from hypoxia in vivo. PMID- 16247768 TI - Cytomorphological alterations of the thymus, spleen, head-kidney, and liver in cardinal fish (Apogonidae, Teleostei) as bioindicators of stress. AB - Morphological and cytological alterations at the light microscope (LM) and transmission electron microscope (TEM) levels were observed in the thymus, spleen, head-kidney, and liver of cardinal fishes (Apogonidae, Teleostei) from the Gulf of Aqaba, Red Sea, sampled from a strongly polluted site at the northern end of the gulf, and compared to similar samples from a clean, reference site. At the polluted site, the most prominent change was the formation of numerous deposits of cells rich in phagosomes with lipofucin, melanin granules, and phagocytosed debris, including a high increase in number and dimensions of Hassall's corpuscles and melano-macrophage centers. The number of Hassall's corpuscles was 20 (+/-8.0)/mm(2) and of melano-macrophage centers 18 (+/ 4.0)/mm(2) at the polluted site, and 7.0 (+/-4.0)/m(2) vs. 5.0 (+/-2.0)/mm(2) respectively at the reference site. In numerous instances the head kidney's melano-macrophage centers in fishes from the polluted site were encapsulated by reticulocytes, a phenomenon recognized as a marker of neoplasmosis and possible malignancy. In the spleens of fishes from the polluted site, numerous deposits of cell debris, peroxisomes, and enlarged lysosomes were also observed. The livers (hepatopancreas) of fishes from polluted waters demonstrated very strong hyperlipogeny. Many of their hepatocytes were laden with lipid vesicles, fragmented endoplasmic reticulula, and aberrant mitochondria. Although the observed alterations in the glands and liver do not indicate any immediate threat to the life of the fish, they can become crucial with respect to energy turnover and fecundity trajectories. This study strongly suggests the use of cytological alterations in vital organs, such as were observed, as pathological biomarkers to environmental stress. PMID- 16247769 TI - Expression and functional analysis of genes deregulated in mouse placental overgrowth models: Car2 and Ncam1. AB - Different causes, such as maternal diabetes, cloning by nuclear transfer, interspecific hybridization, and deletion of some genes such as Esx1, Ipl, or Cdkn1c, may underlie placental overgrowth. In a previous study, we carried out comparative gene expression analysis in three models of placental hyperplasias, cloning, interspecies hybridization (IHPD), and Esx1 deletion. This study identified a large number of genes that exhibited differential expression between normal and enlarged placentas; however, it remained unclear how altered expression of any specific gene was related to any specific placental phenotype. In the present study, we focused on two genes, Car2 and Ncam1, which both exhibited increased expression in interspecies and cloned hyperplastic placentas. Apart from a detailed expression analysis of both genes during normal murine placentation, we also assessed morphology of placentas that were null for Car2 or Ncam1. Finally, we attempted to rescue placental hyperplasia in a congenic model of IHPD by decreasing transcript levels of Car2 or Ncam1. In situ analysis showed that both genes are expressed mainly in the spongiotrophoblast, however, expression patterns exhibited significant variability during development. Contrary to expectations, homozygous deletion of either Car2 or Ncam1 did not result in placental phenotypes. However, expression analysis of Car3 and Ncam2, which can take over the function of Car2 and Ncam1, respectively, indicated a possible rescue mechanism, as Car3 and Ncam2 were expressed in spongiotrophoblast of Car2 and Ncam1 mutant placentas. On the other hand, downregulation of either Car2 or Ncam1 did not rescue any of the placental phenotypes of AT24 placentas, a congenic model for interspecies hybrid placentas. This strongly suggested that altered expression of Car2 and Ncam1 is a downstream event in placental hyperplasia. PMID- 16247770 TI - Reciprocal expression of lin-41 and the microRNAs let-7 and mir-125 during mouse embryogenesis. AB - In C. elegans, heterochronic genes control the timing of cell fate determination during development. Two heterochronic genes, let-7 and lin-4, encode microRNAs (miRNAs) that down-regulate a third heterochronic gene lin-41 by binding to complementary sites in its 3'UTR. let-7 and lin-4 are conserved in mammals. Here we report the cloning and sequencing of mammalian lin-41 orthologs. We find that mouse and human lin-41 genes contain predicted conserved complementary sites for let-7 and the lin-4 ortholog, mir-125, in their 3'UTRs. Mouse lin-41 (Mlin-41) is temporally expressed in developing mouse embryos, most dramatically in the limb buds. Mlin-41 is down-regulated during mid-embryogenesis at the time when mouse let-7c and mir-125 RNA levels are up-regulated. Our results suggest that mammalian lin-41 is temporally regulated by miRNAs in order to direct key developmental events such as limb formation. PMID- 16247771 TI - Subgroups of Alzheimer's disease based on cerebrospinal fluid molecular markers. AB - Alzheimer's disease, the most common cause of dementia, is multifactorial and heterogeneous; its diagnosis remains probable. We postulated that more than one disease mechanism yielded Alzheimer's histopathology, and that subgroups of the disease might be identified by the cerebrospinal fluid (CSF) levels of proteins associated with senile (neuritic) plaques and neurofibrillary tangles. We immunoassayed levels of tau, ubiquitin, and Abeta(1-42) in retrospectively collected CSF samples of 468 clinically diagnosed Alzheimer's disease patients (N = 353) or non-Alzheimer's subjects (N = 115). Latent profile analysis assigned each subject to a cluster based on the levels of these molecular markers. Alzheimer's disease was subdivided into at least five subgroups based on CSF levels of Abeta(1-42), tau, and ubiquitin; each subgroup presented a different clinical profile. These subgroups, which can be identified by CSF analysis, might benefit differently from different therapeutic drugs. PMID- 16247772 TI - Determining particle density distribution of expanded bed adsorbents. AB - This study presents an experimental approach to measure the density distribution of expanded bed adsorption (EBA) matrices. We report on the use of a series of solutions of caesium trifluoroacetate (CsTFA) of varying density spun in a laboratory centrifuge so as to separate representative matrix samples on the basis of bead density. Mass data was used to plot a decumulative density distribution for the matrix. By performing laser light scattering-based measurements on the same samples of matrix the variation in particle size with density was determined. Particle settling velocity distributions were then calculated using these data and compared with a settling velocity distribution calculated on the basis of an assumed constant bead density. The study demonstrates a reliable and simple method for the characterisation of matrix density distribution. For the case of the Streamline matrices tested the particle size distribution is constant with varying bead density. Bead densities varied from 1.5 to 2.1 g/cm3 in the CsTFA solutions. These were then adjusted using bead porosity to give a density range of 1.11-1.33 g/cm3 in aqueous buffer (assumed 1.0 g/cm3) The differences in resultant settling velocity distributions when based upon measured density distribution than when based upon an assumed mean density value were shown to be insignificant. This result confirms experimentally that an assumption of a single constant mean density for EBA particles is acceptable for hydrodynamic modelling and performance prediction purposes. PMID- 16247773 TI - A mathematical model of N-linked glycosylation. AB - Metabolic engineering of N-linked oligosaccharide biosynthesis to produce novel glycoforms or glycoform distributions of a recombinant glycoprotein can potentially lead to an improved therapeutic performance of the glycoprotein product. A mathematical model for the initial stages of this process, up to the first galactosylation of an oligosaccharide, was previously developed by Umana and Bailey (1997) (UB1997). Building on this work, an extended model is developed to include further galactosylation, fucosylation, extension of antennae by N acetyllactosamine repeats, and sialylation. This allows many more structural features to be predicted. A number of simplifying assumptions are also relaxed to incorporate more variables for the control of glycoforms. The full model generates 7565 oligosaccharide structures in a network of 22,871 reactions. Methods for solving the model for the complete product distribution and adjusting the parameters to match experimental data are also developed. A basal set of kinetic parameters for the enzyme-catalyzed reactions acting on free oligosaccharide substrates is obtained from the previous model and existing literature. Enzyme activities are adjusted to match experimental glycoform distributions for Chinese Hamster Ovary (CHO). The model is then used to predict the effect of increasing expression of a target glycoprotein on the product glycoform distribution and evaluate appropriate metabolic engineering strategies to return the glycoform profile to its original distribution pattern. This model may find significant utility in the future to predict glycosylation patterns and direct glycoengineering projects to optimize glycoform distributions. PMID- 16247774 TI - Optimization of DsRed production in Escherichia coli: effect of ribosome binding site sequestration on translation efficiency. AB - DsRed-Express, a popular reporter protein, cannot be expressed in Escherichia coli using a consensus ribosome binding site (RBS) potentially due to basepairing in the RBS that inhibits translation initiation. Saturation mutagenesis was used to probe for a gene sequence that minimized basepairing in the RBS while maintaining the same spectral properties and maturation characteristics as DsRed Express. The new DsRed, designated here as RFP(EC) for E. coli optimized red fluorescent protein, fluoresces 2.5 times greater than DsRed-Express when expressed from the same vector. PMID- 16247775 TI - Gli3 null mice display glandular overgrowth of the developing stomach. AB - The role of the Hedgehog signaling pathway in various aspects of gut development is still poorly understood. In the developing stomach, Sonic (Shh) and Indian (Ihh) hedgehog are expressed in both distinct and overlapping regions. Loss of Sonic hedgehog function in the stomach results in a glandular phenotype of intestinal transformation and overgrowth. These changes are reminiscent of the pre-malignant lesion, intestinal metaplasia. To determine the role of Hedgehog related transcription factors, Gli2 and Gli3, in Shh signaling during stomach development, we conducted a mutant analysis of glandular stomach from Shh, Gli2, and Gli3 mutant mice. Although Gli2 principally mediates the activator function of Shh, surprisingly we observed minimal changes in glandular development in the Gli2 mutant stomach. Furthermore, Gli3, which typically functions as a repressor of Hedgehog signal, showed a striking phenocopy of the glandular expansion and intestinal transformation found in Shh mutant stomach. A reduction in apoptotic events was seen in all mutant stomachs with no appreciable changes in proliferation. Both Shh and Gli3 mutant stomachs displayed early changes of intestinal transformation but these did not impact on the overall differentiation of the gastric epithelium. Interestingly, the observation that Gli3 shares a similar glandular phenotype to Shh mutant stomach reveals a possible novel role of Gli3 activator in the developing stomach. The embryonic stomach is a unique model of the Hedgehog pathway function and one that may help to uncover some of the mechanisms underlying the development of intestinal metaplasia. PMID- 16247777 TI - New York AANCART: using participatory research to address the health needs of South Asian and Korean Americans in New York City. AB - Since its inception in April 2000, the New York Asian American Network for Cancer Awareness, Research, and Training (NY AANCART) has conducted cancer control education, research, and infrastructure-building activities in collaboration with the agenda of the national AANCART project, one of the Special Population Networks funded by the National Cancer Institute. In New York, the AANCART program was developed based on the principles of community-based participatory research (CBPR), a research approach that is centered on community strengths and issues and that engages the community in the research process. NY AANCART has been dedicated to developing leadership within and collaboration with community based organizations (CBOs) to address the needs of medically underserved New York Asian-American populations. In particular, NY AANCART has worked with CBOs to develop and implement a diverse public health agenda that addresses the concerns of the communities that are served by these organizations. Using case studies from two community-based partnerships that were developed by NY AANCART, the current report describes how the principles of CBPR were applied in two rapidly growing Asian-American communities in New York City. PMID- 16247776 TI - Assessing the oral cancer risk of South-Asian immigrants in New York City. AB - Increasing immigration from South Asia, where oral cancer is one of the most common cancers in adults, suggests that this disease will probably pose a serious public health problem in the United States. According to the 2000 Census, there are approximately 1.9 million South Asians in the U.S., and their access to dental care is limited. The morbidity and mortality associated with oral cancer can be reduced by primary prevention and early detection. South Asians are not a homogenous group, and areca, and important risk factor for oral cancer, has religious and cultural significance in some South Asian communities. In order to develop culturally relevant preventive and educational interventions, it is important to assess risk behaviors according to ethnic and religious subgroup identity. In this report, the author provides an overview of oral cancer risk factors among South-Asian immigrants in the U.S., and describes a recently funded pilot study designed to assess oral cancer risk behaviors in South Asian seniors. The data collected will be used to develop risk profiles of South Asian religious and ethnic sub-groups. It is hoped that medical and dental providers will be able to use the risk profiles to target early-detection and risk-reduction services in this population. PMID- 16247778 TI - Keynote speech: progress with a purpose. PMID- 16247779 TI - Promoting adjustment after treatment for cancer. AB - The transition from the period of diagnosis and medical treatment of cancer to survivorship (i.e., the reentry phase) is an understudied phase in the cancer trajectory. The objectives of this report were 1) to illustrate several adaptive tasks of the reentry phase, 2) to provide examples of research on factors that predict positive adjustment during this phase, and 3) to discuss interventions that address the adaptive tasks of early cancer survivorship. Although the pertinent empirical literature is scarce, accounts from cancer survivors, healthcare professionals, and qualitative researchers converge to suggest several themes in adaptive tasks during reentry. Drawing from the authors' work and that of others, the authors have described common expectancies held by many individuals approaching reentry (e.g., "I shouldn't need support"), typical concerns during this phase (e.g., concern over cancer recurrence), and personal and contextual factors that can facilitate and hinder adjustment. Promising psychosocial interventions have been developed for individuals in the reentry period. Continued research will be necessary to characterize this important phase of cancer survivorship. PMID- 16247780 TI - Long-term outcomes of adult survivors of childhood cancer. AB - During the past 30 years, changes in the treatment of children and adolescents with cancer have led to substantial improvements in survival. Although treatment related factors have been shown to impact subsequent health status and quality of life, there is limited information on survivors who are now two or more decades after treatment. The Childhood Cancer Survivor Study (CCSS) was established as a resource for investigating the long-term outcomes of a cohort of 5-year survivors of childhood and adolescent cancer, diagnosed between 1970-1986. The CCSS cohort has more than 14,000 active participants, including survivors of leukemia, brain tumors, Hodgkin disease, non-Hodgkin lymphoma, Wilms tumor, neuroblastoma, soft tissue sarcoma, and bone tumors. Study participants, extensively characterized by their cancer therapy, have provided self-reported sociodemographic- and health related information. Although the survivor population has been found to be at significantly increased risk of several adverse outcomes, such as late mortality, second cancers, pulmonary complications, pregnancy loss, low birth weight of offspring, and decreased education, the overall proportion of survivors affected is relatively small. Subgroups at high risk of adverse outcomes, defined by treatment-related, demographic, or medical factors, can be identified. The ongoing evaluation of large and diverse cohorts of cancer survivors will aid in further identifying individuals who should be the target of innovative intervention strategies. PMID- 16247781 TI - Taking positive changes seriously. AB - It is a common finding that 30-90% of people experiencing serious illness and other forms of adversity report increased quality of life and other positive life changes after their experience, yet there is still considerable disagreement among scientists and practitioners about whether such changes are real or beneficial. The current article identified four pervasive assumptions about positive emotions and beliefs that may limit progress in understanding and promoting resilience among survivors of cancer, namely, that positive beliefs and emotions are absent, dangerous, delusional, or trivial among people managing life threatening illnesses. A review of evidence on each of these points suggested instead that positive emotions and beliefs seem not only to be associated with good outcomes among people experiencing adversity, but also to play a role in realizing them. Implications for the study of positive beliefs, emotions, and life changes among survivors of cancer and for interventions to promote mental and physical health were discussed. PMID- 16247782 TI - Support groups in advanced breast cancer. AB - Considerable research has been conducted into the potential benefits of support groups for patients with metastatic breast cancer. An early report by Spiegel et al. suggested that participation in these groups not only had psychologic benefits but also resulted in prolongation of survival. A review of the published literature was undertaken. Five randomized trials of support groups in metastatic breast cancer have been published. Four of those five trials have reported survival results: Survival outcomes from the fifth study are pending. A variety of group interventions were investigated. All of the studies reported beneficial psychosocial effects of the intervention, although the effects, at times, were transient and were not always present for all study outcomes. Only the original publication by Spiegel's group identified a survival benefit. No survival benefit was seen for participation in support groups for the other three studies that reported survival effects. Mixed survival effects were reported for other psychologic interventions in other cancers. There was good evidence that support groups in metastatic breast cancer lead to improved psychologic outcomes. Evidence of beneficial survival effects was not convincing. PMID- 16247783 TI - Neural correlates of anosognosia for cognitive impairment in Alzheimer's disease. AB - We explored the neural substrate of anosognosia for cognitive impairment in Alzheimer's disease (AD). Two hundred nine patients with mild to moderate dementia and their caregivers assessed patients' cognitive impairment by answering a structured questionnaire. Subjects rated 13 cognitive domains as not impaired or associated with mild, moderate, severe, or very severe difficulties, and a sum score was calculated. Two measures of anosognosia were derived. A patient's self assessment, unconfounded by objective measurements of cognitive deficits such as dementia severity and episodic memory impairment, provided an estimate of impaired self-evaluative judgment about cognition in AD. Impaired self-evaluation was related to a decrease in brain metabolism measured with 18F-2 fluoro-2-deoxy-D-glucose positron emission tomography (FDG-PET) in orbital prefrontal cortex and in medial temporal structures. In a cognitive model of anosognosia, medial temporal dysfunction might impair a comparison mechanism between current information on cognition and personal knowledge. Hypoactivity in orbitofrontal cortex may not allow AD patients to update the qualitative judgment associated with their impaired cognitive abilities. Caregivers perceived greater cognitive impairments than patients did. The discrepancy score between caregiver's and patient's evaluations, an other measure of anosognosia, was negatively related to metabolic activity located in the temporoparietal junction, consistent with an impairment of self-referential processes and perspective taking in AD. PMID- 16247784 TI - MEG reveals different contributions of somatomotor cortex and cerebellum to simple reaction time after temporally structured cues. AB - Magnetoencephalography (MEG) was used to measure brain activity while participants performed a simple reaction to targets after either a random interval (uncued targets) or a series of isochronous warning stimuli with 200-ms intervals that acted as a countdown. Targets could arrive "on time" or "early" relative to the preceding warning stimuli. Cerebellar activity before any stimulus onset predicted uncued simple reaction time. Onset of activity in somatomotor cortex relative to the target predicted reaction time after two warning stimuli when the target arrived on time or early. After three warning stimuli, when the target arrived on time and was certain to occur, prestimulus cerebellar activity and somatomotor onset were significant predictors of reaction time. When the target arrived early after three warning stimuli, prestimulus cerebellar and cingulate activity were predictive. The cerebellar results may reflect a number of possible factors, including a role in timing, response readiness, prediction and attention. PMID- 16247785 TI - Enhancing adaptation during treatment and the role of individual differences. AB - With increased effectiveness of medical treatment for cancer, greater attention has been directed to the quality of life of cancer patients and cancer survivors. Work on this topic is aimed at better understanding the determinants of quality of life and creating ways to optimize these outcomes. Insights from many disciplines inform this effort. For example, personality psychology suggests bases for naturally occurring differences in resilience but also suggests ways to enhance adaptation and promote greater resilience. The author's current work relies on a view of individuals as goal-seeking beings whose efforts toward desired outcomes are threatened by diagnosis and treatment of cancer. Some react to such adversity with increased efforts, others with a giving-up response. This difference promotes differences in emotional well being. The author's group is focused on enhancing adaptation and providing patients with skills for stress management that permit patients to remain engaged in the pursuits that form their lives. This stress-management intervention has reduced the prevalence of depression among breast cancer patients and has increased the extent to which they report positive sequelae from cancer. The latter effect also has been related to differential reduction in cortisol. Ongoing work is examining well being in longer term survivors, including the effects of this intervention and the possibility that the intervention will have positive consequences for physical health. PMID- 16247786 TI - Electrostatic potential at the retinal of three archaeal rhodopsins: implications for their different absorption spectra. AB - The color tuning mechanism of the rhodopsin protein family has been in the focus of research for decades. However, the structural basis of the tuning mechanism in general and of the absorption shift between rhodopsins in particular remains under discussion. It is clear that a major determinant for spectral shifts between different rhodopsins are electrostatic interactions between the chromophore retinal and the protein. Based on the Poisson-Boltzmann equation, we computed and compared the electrostatic potential at the retinal of three archaeal rhodopsins: bacteriorhodopsin (BR), halorhodopsin (HR), and sensory rhodopsin II (SRII) for which high-resolution structures are available. These proteins are an excellent test case for understanding the spectral tuning of retinal. The absorption maxima of BR and HR are very similar, whereas the spectrum of SRII is considerably blue shifted--despite the structural similarity between these three proteins. In agreement with their absorption maxima, we find that the electrostatic potential is similar in BR and HR, whereas significant differences are seen for SRII. The decomposition of the electrostatic potential into contributions of individual residues, allowed us to identify seven residues that are responsible for the differences in electrostatic potential between the proteins. Three of these residues are located in the retinal binding pocket and have in fact been shown to account for part of the absorption shift between BR and SRII by mutational studies. One residue is located close to the beta-ionone ring of retinal and the remaining three residues are more than 8 A away from the retinal. These residues have not been discussed before, because they are, partly because of their location, no obvious candidates for the spectral shift among BR, HR, and SRII. However, their contribution to the differences in electrostatic potential is evident. The counterion of the Schiff base, which is frequently discussed to be involved in the spectral tuning, does not contribute to the dissimilarities between the electrostatic potentials. PMID- 16247787 TI - Remarks of Ellen Stovall NCI/ACS Survivorship Symposium June 2002. PMID- 16247788 TI - Cognitive functioning after adjuvant chemotherapy and/or radiotherapy for early stage breast carcinoma. AB - BACKGROUND: Evidence suggests that women diagnosed with early-stage breast carcinoma may experience cognitive problems as a consequence of adjuvant chemotherapy treatment. The present study was conducted to examine whether there are differences in cognitive performance and cognitive complaints between women treated with and without chemotherapy for TNM Stage 0 to II breast carcinoma. METHODS: As part of a larger study on quality of life, women were recruited with newly diagnosed Stage 0 to II breast carcinoma scheduled to be treated with chemotherapy plus radiotherapy (n = 60) or radiotherapy only (n = 83). Six months after the completion of treatment, participants were administered a standard neuropsychologic battery to assess cognitive performance and a self-report measure to assess perceived cognitive problems. RESULTS: There were no statistically significant differences between women who received chemotherapy and those who did not with regard to their average performance on tests of episodic memory, attention, complex cognition, motor performance, or language. Likewise, there were no significant differences between the treatment groups in the prevalence of impairment in each of these cognitive domains. Women who underwent chemotherapy also did not report significantly more problems with cognitive functioning than women treated without chemotherapy. CONCLUSIONS: The findings failed to confirm previous reports suggesting adjuvant chemotherapy is associated with problems in cognitive functioning among women who receive treatment for Stage 0 to II breast carcinoma. Future research should use prospective longitudinal research designs incorporating appropriate comparison groups to further explore this issue. PMID- 16247790 TI - Notable effects of angiotensin II receptor blocker, valsartan, on acute cardiotoxic changes after standard chemotherapy with cyclophosphamide, doxorubicin, vincristine, and prednisolone. AB - BACKGROUND: There are three distinct types of doxorubicin-induced cardiotoxicity (acute, chronic, and late-onset). Although previous studies with animal models suggest that angiotensin II plays a key role in the process of the doxorubicin induced cardiotoxicity, there has been no such observation in humans. This randomized study investigated whether valsartan, a new class of angiotensin II receptor blocker (ARB), can inhibit acute cardiotoxicity after doxorubicin-based chemotherapy. METHODS: Forty consecutive patients with untreated non-Hodgkin lymphoma who were scheduled to undergo standard chemotherapy with cyclophosphamide, doxorubicin, vincristine, and prednisolone (CHOP) (mean age, 56 yrs; range, 24-70 yrs) were randomized with minimization methods to receive CHOP with or without 80 mg/day of valsartan. Acute cardiotoxicity was comprehensively evaluated with neurohumoral, echocardiographic, and electrocardiographic markers before and on Days 3, 5, and 7 after the initiation of CHOP. RESULTS: CHOP induced transient increases in the left ventricular end-diastolic diameter in an echocardiogram, the QTc interval and QTc dispersion in an electrocardiogram, and in the plasma brain and atrial natriuretic peptides. All these changes returned to nearly normal levels within a week after CHOP (P < 0.001). Notably, valsartan significantly prevented all these changes except for the elevation in atrial natriuretic peptide (P < 0.05). No significant change was observed in blood pressure or heart rate between the valsartan and control groups. CONCLUSIONS: The results indicate that angiotensin II may play an essential role in acute CHOP induced cardiotoxicity in humans. Future long-term studies are necessary to judge whether ARBs have a potential to prevent the chronic or late-onset types of doxorubicin-induced cardiotoxicity. PMID- 16247791 TI - The impact of the fourth disulfide bridge in scorpion toxins of the alpha-KTx6 subfamily. AB - Animal toxins are highly reticulated and structured polypeptides that adopt a limited number of folds. In scorpion species, the most represented fold is the alpha/beta scaffold in which an helical structure is connected to an antiparallel beta-sheet by two disulfide bridges. The intimate relationship existing between peptide reticulation and folding remains poorly understood. Here, we investigated the role of disulfide bridging on the 3D structure of HsTx1, a scorpion toxin potently active on Kv1.1 and Kv1.3 channels. This toxin folds along the classical alpha/beta scaffold but belongs to a unique family of short-chain, four disulfide bridged toxins. Removal of the fourth disulfide bridge of HsTx1 does not affect its helical structure, whereas its two-stranded beta-sheet is altered from a twisted to a nontwisted configuration. This structural change in HsTx1 is accompanied by a marked decrease in Kv1.1 and Kv1.3 current blockage, and by alterations in the toxin to channel molecular contacts. In contrast, a similar removal of the fourth disulfide bridge of Pi1, another scorpion toxin from the same structural family, has no impact on its 3D structure, pharmacology, or channel interaction. These data highlight the importance of disulfide bridging in reaching the correct bioactive conformation of some toxins. PMID- 16247792 TI - Asian subgroups and cancer incidence and mortality rates in California. AB - The objective of this study was to characterize better the cancer burden among Asian subgroups in California. Nearly 3.7 million Asians reside in California, and no other state has as many Asians. Cancer statistics for Asians often are combined with statistics for Pacific Islanders, and rates for subgroups are not often examined, because most states do not have a large enough population. Asians are affected disproportionately by certain cancers, such as stomach and liver cancers. The California Cancer Registry, a population-based cancer registry, has collected data, including race/ethnicity data, since 1988. The 5-year, average, annual, age-adjusted cancer incidence and mortality rates from 1997 through 2001 were calculated for 5 Asian subgroups: Chinese, Filipino, Japanese, Korean, and Vietnamese. Cancer incidence and mortality varied greatly. Incidence rates for all sites combined among males varied from a low of 318.6 per 100,000 for Chinese to a high of 366.0 per 100,000 among Japanese. For females, rates ranged from 236.6 per 100,000 among Koreans to 302.4 per 100,000 among Japanese. Mortality rates also varied by Asian subgroup. Presenting one statistic for Asian/Pacific Islanders did not provide an accurate depiction of the cancer burden among the different Asian subgroups. Acculturation will continue to affect the patterns of cancer incidence among Asian subgroups in California. PMID- 16247793 TI - Cancer incidence in the Hmong in California, 1988-2000. AB - The Hmong represent a unique new Southeast Asian immigrant group to the U.S. Approximately 169,000 Hmong reside in the U.S., primarily in California, Minnesota, and Wisconsin. Previous studies of cancer in this population have indicated that Hmong experience an elevated risk of gastric, hepatic, cervical, and nasopharyngeal cancers and experience a reduced risk of breast, prostate, lung, and colorectal cancers. Approximately 65,000 Hmong live in California, where there has been a population-based cancer registry since 1988, and the authors used these data to calculate age-adjusted cancer incidence rates and to examine disease stage and tumor grade at diagnosis. Changes in rates during the period studied also were evaluated. These rates and proportions were compared with rates among the non-Hispanic white (NHW) and Asian/Pacific Islander (API) populations of California. Between 1988 and 2000, a total of 749 Hmong in California were diagnosed with invasive cancer, and the age-adjusted rate of cancer for the Hmong was 284 per 100,000 population, compared with 362.6 and 478 per 100,000 in the API and NHW populations, respectively. The age-adjusted incidence rates of cancer in the Hmong were elevated for hepatic, gastric, cervical, and nasopharyngeal cancers and for leukemia and non-Hodgkin lymphoma (NHL). Rates were lower in the Hmong for colorectal, lung, breast, and prostate cancers. For gastric cancer and lung cancer, age-adjusted rates increased between 1988 and 2000 in the Hmong, although breast cancer incidence declined. Cervical cancer incidence increased, rates of NHL were declining, and rates for colorectal cancer remained steady between 1988 and 2000. The Hmong experienced later disease stage at diagnosis than other API and generally poorer grade of disease at diagnosis. Hmong experienced lower overall invasive cancer incidence rates than API or NHW populations in California. However, they experienced higher rates of hepatic, gastric, cervical, and nasopharyngeal cancers; and, for most types of cancer, they were diagnosed in a later disease stage. PMID- 16247794 TI - Cell transplantation in wounded mixed connective tissues. AB - Direct transplantation of multipotent precursor cells into the periodontium could provide a therapeutic approach for restoring periodontal tissues destroyed by periodontitis or trauma. To improve the understanding of cell migration, proliferation, and differentiation, we used a rodent model combining orthodontic tooth movement and transplantation of Lac-Z-positive murine-cultured periodontal ligament (PL) or femur-derived bone marrow precursor cells into a defined mandibular wound site, thus promoting tissue regeneration in wounded periodontium. Our results show that in orthodontically traumatized tissues, transplanted PL and bone marrow cells migrated systemically, contributing to the repopulation of sites with reduced cell/matrix density. The transplanted PL cells proliferated in adjacent alveolar bone marrow spaces, thus migrating to vascular tissues in the PL. The capillary walls in the PL serve as delivery sites for these cells and other marrow-derived hematopoietic cells, including monocytes. The transplanted marrow cells, extracted from femur of transgenic (TgR) mice exhibited similar behavior to those of transplanted PL cells, showing high proliferative activity in alveolar marrow as well as intensive repopulating capacity in wounded periodontium. On the other hand, the buccal skin fibroblasts failed to migrate and home effectively and thus the transplantation of these cells had no effect on periodontium regeneration. Based on these results, we conclude that the transplanted PL and bone marrow cells migrate systemically and following a cyclical process of growth and development and differentiate into PL fibroblasts, osteoblasts, and cementoblasts, thereby contributing to periodontal regeneration. PMID- 16247796 TI - Clinical trials: understanding and perceptions of female Chinese-American cancer patients. AB - Under-representation of minority and female participants prompted the U.S. legislature to mandate the inclusion of women and minorities in federally funded research. Recruitment of minorities to participate in clinical trials continues to be challenging. Although Asian Americans constitute one of the major minority groups in the U.S., published literature contains sparse data concerning the participation of Asian Americans in cancer clinical trials. The authors completed qualitative, semistructured interviews with 34 participants: Chinese-American female cancer patients ages 20-85 years or their family members. Interviews were conducted in Cantonese, Mandarin, or English and were audiotaped. Chinese interviews were translated into English, and all interviews were transcribed subsequently into English. A team of five coders individually reviewed then met to discuss the English transcripts. The authors used the constant comparative technique throughout the entire coding process as part of the analysis. Among participants, 62% lacked any knowledge of clinical trials, and many expressed negative attitudes toward clinical trials. Barriers to participation included inadequate resources, language issues, and a lack of financial and social support. Facilitating factors included recommendations by a trusted oncologist or another trusted individual and information in the appropriate language. It is noteworthy that family members played an important role in the cancer experience of these participants. To promote participation, there is a need to increase knowledge of clinical trials among Chinese cancer patients. It also is necessary to examine the applicability of current patient-physician communication and interaction models. In addition, decision-making based on Asian philosophies within the context of Euro-American bioethics requires further study. PMID- 16247795 TI - Asian Americans and cancer clinical trials: a mixed-methods approach to understanding awareness and experience. AB - Cancer clinical trials have been based on low accrual rates. Barriers to recruitment of minority populations affect the generalizability and impact of trial findings for those populations. The authors undertook a mixed-methods approach to understanding levels of awareness and experiences with cancer clinical trials. A survey was administered to new cancer patients and their caretakers (family, close friends, or other social support) at outpatient oncology clinics. Field observations of the trial accrual process also were conducted by employing the grounded theory approach in qualitative methods. Comparison of survey results for Asian-American respondents and non-Asian respondents indicated that Asians were less likely to have heard the term "clinical trial" and were more likely to define a clinical trial as "an experiment" or "a test procedure in a clinic" than non-Asians. Asians were more likely to have employer-based insurance and to report understanding issues related to cost reimbursement. Asians were less likely to have been involved in or to know someone in a trial and reported less willingness than white respondents to consider trial participation. Qualitative observations suggested that Asians who presented for a potential trial were interested in the availability of a novel cancer therapy but were not eligible for available trials. Multiple strategies will be necessary to enhance awareness of and experience with accrual to cancer clinical trials for Asians, including richer understanding and increased involvement of Asians in cancer clinical trials and greater attention to the location and diversity of the Asian population in structuring study centers and evaluating trial results. PMID- 16247797 TI - Human BMP-2 gene transfer using transcutaneous in vivo electroporation induced both intramembranous and endochondral ossification. AB - It has been generally accepted that bone morphogenetic protein-2 (BMP-2) can induce osteogenesis in skeletal muscles via endochondral ossification. However, it is not clear how the ossification process occurs after the BMP-2 gene transfer to skeletal muscles in rats using in vivo electroporation. In this study, we evaluated the ossification process by BMP-2 gene transfer using in vivo electroporation. The gastrocnemius muscles of Wistar rats were injected with human BMP-2 gene expression vector (pCAGGS-BMP-2), followed by electroporation under the condition of 100 V, 50 msec per 1 sec, x8. Light and electron microscopic and radiographic analyses were performed at 1, 3, 5, 7, and 10 days after treatment. At 7 days, no sign of cartilage and/or bone formation was detected. However, at 10 days after in vivo electroporation, soft X-ray analysis revealed small lucent areas around the plasmid-injected region. Clusters of both cartilage tissues, leading to endochondral ossification and intramembranous bones of various sizes, were observed between muscle fibers. RT-PCR detected osteocalcin mRNA, showing bone formation at 10 days. Our findings strongly suggest that BMP-2 gene transfer using in vivo electroporation induces not only endochondral ossification but also intramembranous ossification. PMID- 16247798 TI - Survey of the geometric association of domain-domain interfaces. AB - Considering the limited success of the most sophisticated docking methods available and the amount of computation required for systematic docking, cataloging all the known interfaces may be an alternative basis for the prediction of protein tertiary and quaternary structures. We classify domain interfaces according to the geometry of domain-domain association. By applying a simple and efficient method called "interface tag clustering," more than 4,000 distinct types of domain interfaces are collected from Protein Quaternary Structure Server and Protein Data Bank. Given a pair of interacting domains, we define "face" as the set of interacting residues in each single domain and the pair of interacting faces as an "interface." We investigate how the geometry of interfaces relates to a network of interacting protein families, such as how many different binding orientations are possible between two families or whether a family uses distinct surfaces or the same surface when the family has diverse interaction partners from various families. We show there are, on average, 1.2 1.9 different types of interfaces between interacting domains and a significant number of family pairs associate in multiple orientations. In general, a family tends to use distinct faces for each partner when the family has diverse interaction partners. Each face is highly specific to its interaction partner and the binding orientation. The relative positions of interface residues are generally well conserved within the same type of interface even between remote homologs. The classification result is available at http://www.biotec.tu dresden.de/~wkim/supplement. PMID- 16247799 TI - Structural basis of the substrate subsite and the highly thermal stability of xylanase 10B from Thermotoga maritima MSB8. AB - The crystal structure of xylanase 10B from Thermotoga maritima MSB8 (TmxB), a hyperthermostable xylanase, has been solved in its native form and in complex with xylobiose or xylotriose at 1.8 A resolution. In order to gain insight into the substrate subsite and the molecular features for thermal stability, we compared TmxB with family 10 xylanase structures from nine microorganisms. As expected, TmxB folds into a (beta/alpha)8-barrel structure, which is common among the glycoside hydrolase family 10. The enzyme active site and the environment surrounding the xylooligosaccharide of TmxB are highly similar to those of family 10 xylanases. However, only two xylose moieties were found in its binding pocket from the TmxB-xylotriose complex structure. This finding suggests that TmxB could be a potential biocatalyst for the large-scale production of xylobiose. The result of structural analyses also indicated that TmxB possesses some additional features that account for its thermostability. In particular, clusters of aromatic residues together with a lack of exposed hydrophobic residues are characteristic of the TmxB structure. TmxB has also a significant number of ion pairs on the protein surface that are not found in other thermophilic family 10 xylanases. PMID- 16247800 TI - Tobacco cembranoids protect the function of acute hippocampal slices against NMDA by a mechanism mediated by alpha4beta2 nicotinic receptors. AB - Nicotine has been reported to be neuroprotective in experimental and epidemiological studies. In addition to nicotine, tobacco and cigarette smoke contain cembranoids, which are antagonists of neuronal nicotinic receptors (nAChR). Exposure of hippocampal slices to N-methyl-D-aspartate (NMDA) decreases the population spikes (PS). This parameter has been used as a measure of excitotoxicity. Surprisingly, both nicotine and tobacco cembranoids protected against NMDA and this neuroprotection was not blocked by methyllycaconitine (MLA), an antagonist of alpha7 nAChR. On the contrary, MLA had a neuroprotective effect of its own. We examined the effect of the tobacco cembranoid (1S,2E,4R,6R,7E,11E)-cembra-2,7,11-triene-4,6-diol (4R) on the neuroprotection against NMDA. DHbetaE, a selective antagonist of alpha4beta2 nAChR, inhibited the neuroprotection by nicotine, 4R, and MLA, suggesting the involvement of alpha4beta2 nAChRs in the neuroprotection. The cell-signaling pathways underlying the neuroprotection by 4R and by nicotine are different. The activity of phosphatidylinositol-3 kinase (PI3K) was required in both cases; however, 4R required the activity of L-type calcium channels and CAM kinase, whereas nicotine required the extracellular signal regulated kinase-1,2 (ERK) and protein kinase C (PKC). In addition, 4R did not enhance total phospho-ERK-1/2 but increased the amount of total Akt/PKB phosphorylated on the activation site and of glycogen synthase kinase 3-beta phosphorylated on the inhibitory site. Total levels of phosphoenzymes are presented instead of the ratio of phospho- over total enzyme because in preliminary experiments total ERK-1/2 levels were slightly increased by 4R. In conclusion, these findings demonstrate that there are two different nicotinic neuroprotective mechanisms mediated by alpha4beta2. PMID- 16247801 TI - Proliferation and migration of granule cells in the developing rat cerebellum: cisplatin effects. AB - We evaluated the relationship among proliferation, death and migration of granule cells in lobules VI-VIII of vermis, in comparison with lobule III, during cerebellar development. To this aim, a single injection of cisplatin, i.e., a cytostatic agent that is known to induce death of proliferating granule cells, was given to 10-day-old rats. Histochemical markers of proliferating (PCNA immunoreaction) and apoptotic (TUNEL staining) cells were used; the variations of the external granular layer (EGL) thickness were evaluated in parallel. After PCNA and TUNEL reactions, evident changes of the whole EGL were found on PD11 (1 day after treatment), when a reduction of the thickness of this layer was found in treated rats, mainly in consequence of the high number of apoptotic cells in all the cerebellar lobules. On PD17 (7 days after treatment), a thick layer of proliferating cells was observed in lobules VI-VIII of treated rats, while the peculiar pattern of the normal development showed a thin EGL. At the same time, in treated rats, the number of apoptotic cells in EGL was low. In all developmental stages of treated rats, after GFAP immunoreaction, glial fibers appeared twisted, thickened, and with an irregular course; intensely labeled end feet were present. The damage of radial glia suggests an alteration of migratory processes of granule cells, which is also evidenced by the decreased thickness of the premigratory zone of the EGL. Injured radial glia fibers were restricted to lobules VI-VIII and they persisted at PD30, leading to the presence of ectopic granule cells in the molecular layer, as we previously described. PMID- 16247802 TI - Postsynaptic excitation of prefrontal cortical pyramidal neurons by hypocretin 1/orexin A through the inhibition of potassium currents. AB - Hypocretins are crucial for the regulation of wakefulness by the excitatory actions on multiple subcortical arousal systems. To date, there is little information about the direct postsynaptic excitatory effects of hypocretins on the neurons in prefrontal cortex (PFC), which is important for higher cognitive functions and is correlated with level of wakefulness. In this study, we tested the excitatory effects of hypocretin-1 on acutely isolated PFC pyramidal neurons of rats and studied the possible ionic mechanisms by using whole-cell patch-clamp techniques. Puff application of hypocretin-1 caused a dose-dependent excitation. Further observations that perfusion of Ca2+-free artificial cerebrospinal fluid did not influence the depolarizing effects of hypocretin-1, in conjunction with the findings that hypocretin-1 could decrease net whole-cell K+ currents, demonstrate that the excitatory effects of hypocretin-1 on PFC neurons are mediated by the inhibition of K+ currents but not Ca2+ influx. Finally, the decrease in K+ currents induced by hypocretin-1 was abolished by a protein kinase C (PKC) inhibitor (BIS II) or a phospholipase C (PLC) inhibitor (D609), suggesting that PKC and PLC appear to be involved in mediating the inhibitory effects of hypocretin-1 on K+ currents. These results indicate that hypocretin-1 exerts a postsynaptic excitatory action on PFC neurons through the inhibition of K+ currents, which probably results from activation of PKC and PLC signaling pathways. PMID- 16247803 TI - Transplanted human embryonic germ cell-derived neural stem cells replace neurons and oligodendrocytes in the forebrain of neonatal mice with excitotoxic brain damage. AB - Stem cell therapy is a hope for the treatment of some childhood neurological disorders. We examined whether human neural stem cells (hNSCs) replace lost cells in a newborn mouse model of brain damage. Excitotoxic lesions were made in neonatal mouse forebrain with the N-methyl-D-aspartate (NMDA) receptor agonist quinolinic acid (QA). QA induced apoptosis in neocortex, hippocampus, striatum, white matter, and subventricular zone. This degeneration was associated with production of cleaved caspase-3. Cells immunopositive for inducible nitric oxide synthase were present in damaged white matter and subventricular zone. Three days after injury, mice received brain parenchymal or intraventricular injections of hNSCs derived from embryonic germ (EG) cells. Human cells were prelabeled in vitro with DiD for in vivo tracking. The locations of hNSCs within the mouse brain were determined through DiD fluorescence and immunodetection of human specific nestin and nuclear antigen 7 days after transplantation. hNSCs survived transplantation into the lesioned mouse brain, as evidenced by human cell markers and DiD fluorescence. The cells migrated away from the injection site and were found at sites of injury within the striatum, hippocampus, thalamus, and white matter tracts and at remote locations in the brain. Subsets of grafted cells expressed neuronal and glial cell markers. hNSCs restored partially the complement of striatal neurons in brain-damaged mice. We conclude that human EG cell-derived NSCs can engraft successfully into injured newborn brain, where they can survive and disseminate into the lesioned areas, differentiate into neuronal and glial cells, and replace lost neurons. (c) 2005 Wiley-Liss, Inc. PMID- 16247804 TI - Ischemic preconditioning ameliorates excitotoxicity by shifting glutamate/gamma aminobutyric acid release and biosynthesis. AB - Excitotoxicity is recognized to play a major role in cerebral ischemia-induced cell death. The main goal of the present study was to define whether our model of ischemic preconditioning (IPC) promotes a shift from excitatory to inhibitory neurotransmission during the test ischemia to diminish metabolic demand during the reperfusion phase. We also determined whether gamma-aminobutyric acid (GABA) played a role in IPC-induced neuroprotection. Ten minutes of cerebral ischemia was produced by tightening the carotid ligatures bilaterally following hypotension. Samples of microdialysis perfusate, representing extracellular fluid, were analyzed for amino acid content by HPLC. IPC promoted a robust release of GABA after lethal ischemia compared with control rats. We also observed that the activity of glutamate decarboxylase (the predominant pathway of GABA synthesis in the brain) was higher in the IPC group compared with control and ischemic groups. Because GABAA receptor up-regulation has been shown to occur following IPC, and GABAA receptor activation has been implicated in neuroprotection against ischemic insults, we tested the hypothesis that GABAA or GABAB receptor activation was neuroprotective during ischemia or early reperfusion by using an in vitro model (organotypic hippocampal slice culture). Administration of the GABAB agonist baclofen during test ischemia and for 1 hr of reperfusion provided significant neuroprotection. We concluded that increased GABA release in preconditioned animals after ischemia might be one of the factors responsible for IPC neuroprotection. Specific activation of GABAB receptor contributes significantly to neuroprotection against ischemia in organotypic hippocampal slices. PMID- 16247805 TI - Hormonal and sensory inputs regulate distinct neuroblast cell cycle properties in adult cricket brain. AB - From invertebrates to humans, it has been demonstrated that new neurons are added to specific brain structures throughout adult life. In the house cricket, adult neurogenesis occurs in the mushroom bodies, the main sensory integrative center of the brain, often considered an analogue of vertebrate hippocampus. We have previously shown that this neurogenesis can be modulated by hormones through the polyamine pathway and by environmental conditions through sensory inputs and the nitric oxide pathway. Environment-induced neurogenesis is independent of juvenile hormone levels, so we addressed the roles of sensory inputs and hormones in the control of neuroblast proliferation. Here, by using double labelling of cells specifically in S phase (5-bromo-2'-deoxyuridine) together with labelling of mitotically active cells in any phase (proliferating cell nuclear antigen), we show that juvenile hormone acts on progenitor cell proliferation by inducing quiescent neuroblasts to enter the cell cycle, whereas sensory inputs act by shortening the cell cycle. Thus, in the adult house cricket, regulation of neuroblast proliferation by hormonal and environmental cues occurs through two independent modes of action. PMID- 16247806 TI - Health behaviors during and after a cancer diagnosis. AB - With improvements in cancer survival rates, more patients with cancer are living longer, and hence, cancer is becoming viewed as a chronic illness requiring long term management. An important aspect of patient care during and after cancer treatment is patient health behaviors. For example, sequelae from various cancer treatments, such as chemotherapy and radiotherapy (RT), can compromise health in a variety ways, including decreased immune functioning, cardiotoxic effects of chemotherapy and/or RT, and weight gain. In addition, the stress caused by a cancer diagnosis and its treatment can disrupt existing health behaviors or exacerbate unhealthy behaviors. Continued smoking or alcohol use can complicate treatment and increase risk for further malignancy. Furthermore, decreased physical activity and poor nutrition can cause weight gain, which may contribute to secondary health problems such as cardiovascular disease and diabetes. The authors reviewed the extant literature on four key health behaviors among patients with cancer and survivors: healthy diet, reduced tobacco use, reduced alcohol use, and increased physical activity. They described the prevalence of these behaviors, reviewed the effects of interventions designed to alter unhealthy behaviors, and discussed the implications and future directions for this emerging area of research. PMID- 16247807 TI - Participation of Asian-American women in cancer chemoprevention research: physician perspectives. AB - To the authors' knowledge, little is known regarding the participation of Asian Americans in cancer prevention research. In 2002, the authors mailed surveys to primary care physicians in Northern California to assess their knowledge, attitudes, behaviors, and barriers concerning the participation of Asian-American women in breast cancer chemoprevention research. The response rate was 52.3% (n=306 physicians). For physician barriers, most respondents selected lack of study knowledge (73%) and effort required to establish eligibility (75%) and to explain risks and benefits (68%). For patient barriers, most physicians chose the following: physicians did not inform patients about trials (76%), limited English proficiency (78%), researcher-participant language discordance (74%), and complex protocols (69%). Significantly more Asian-American physicians than non-Asian American physicians (but a majority of each) selected as patient barriers a lack of culturally relevant information on breast cancer, a lack of knowledge about research concepts, and fear of experimentation. A majority of Asian-American physicians also selected the following patient barriers: lack of knowledge of preventive care or breast cancer, work concern, misperception that experimental treatment is inferior, personal modesty, and lack of personal benefit. In multivariate analyses, physicians who were in practice longer, who spent more time with patients, or who knew of tools to estimate breast cancer risk were more likely to discuss such trials with Asian-American women; whereas male physicians and those who believed that Asian-American women's deference to physicians was a barrier were less likely to have discussed such trials with Asian-American women. Efforts to increase research participation among Asian Americans should include physician education and linguistically appropriate recruitment efforts. PMID- 16247808 TI - Lipopolysaccharide and proinflammatory cytokines require different astrocyte states to induce nitric oxide production. AB - Nitric oxide (NO) production by astrocytes is a significant factor affecting brain physiology and pathology, but the mechanism by which it is regulated is not known. Previous studies using different specimens and stimuli might have described different aspects of a complex system. We investigated the effect of culture and stimulus conditions on NO production by cultured astrocytes and identified two combinations of these allowing NO production. Lipopolysaccharide (LPS)-induced NO production required a high seeding cell density and was independent of the serum concentration, whereas that induced by proinflammatory cytokines required simultaneous treatment with interleukin-1beta, tumor necrosis factor-alpha, and interferon-gamma and low-serum conditions but was less affected by the seeding density. These two pathways showed differential sensitivity to protein kinase inhibitors. Both LPS and cytokines induced expression of inducible nitric oxide synthase (iNOS). Although LPS-induced iNOS expression required a high seeding cell density, cytokine-induced iNOS expression, in contrast to NO production, was not affected by the serum concentration. These results suggest that astrocytes interact with the environment and alter their responsiveness to NO production-inducing stimuli by regulating iNOS expression and activity. This is the first evidence for the selective use of two different regulatory pathways in any cell type. PMID- 16247809 TI - Inter- and intrahabitat dietary variability of chacma baboons (Papio ursinus) in South African savannas based on fecal delta13C, delta15N, and %N. AB - Baboons are dietary generalists, consuming a wide range of food items in varying proportions. It is thus difficult to quantify and explain the dietary behavior of these primates. We present stable carbon (delta(13)C) and nitrogen (delta(15)N) isotopic data, and percentage nitrogen (%N), of feces from chacma baboons (Papio ursinus) living in two savanna environments of South Africa: the mountainous Waterberg region and the low-lying Kruger National Park. Baboons living in the more homogeneous landscapes of the Waterberg consume a more isotopically heterogeneous diet than their counterparts living in Kruger Park. Grasses and other C(4)-based foods comprise between approximately 10-20% (on average) of the bulk diet of Kruger Park baboons. Carbon isotopic data from the Waterberg suggest diets of approximately 30-50% grass, which is higher than generally reported for baboons across the African savanna. Based on observations of succulent-feeding, we propose that baboons in the Waterberg consume a mix of C(4) grasses and CAM photosynthesizing succulents in combined proportions varying between approximately 5-75% (average, approximately 35%). Fecal delta(15)N of baboons is lower than that of sympatric ungulates, which may be due to a combination of low levels of faunivory, foraging on subterranean plant parts, or the use of human foods in the case of Kruger Park populations. Fecal N levels in baboons are consistently higher than those of sympatric ungulate herbivores, indicating that baboons consume a greater proportion of protein-rich foods than do other savanna mammals. These data suggest that chacma baboons adapt their dietary behavior so as to maximize protein intake, regardless of their environment. PMID- 16247810 TI - Cationic yttrium methyl complexes as functional models for polymerization catalysts of 1,3-dienes. PMID- 16247811 TI - Large-scale nonhydrolytic sol-gel synthesis of uniform-sized ceria nanocrystals with spherical, wire, and tadpole shapes. PMID- 16247812 TI - Quantum dots as efficient energy acceptors in a time-resolved fluoroimmunoassay. PMID- 16247814 TI - Synthetic vaccines consisting of tumor-associated MUC1 glycopeptide antigens and bovine serum albumin. PMID- 16247813 TI - Oriented nanoporous lamellar organosilicates templated from topologically unsymmetrical dendritic-linear block copolymers. PMID- 16247815 TI - A fully synthetic vaccine consisting of a tumor-associated glycopeptide antigen and a T-cell epitope for the induction of a highly specific humoral immune response. PMID- 16247816 TI - A highly active anion-selective aminocyclodextrin ion channel. PMID- 16247817 TI - Stereoselective palladium-catalyzed four-component cascade synthesis of pyrrolidinyl-, pyrazolidinyl-, and isoxazolidinyl isoquinolines. PMID- 16247818 TI - Oxygen storage at the metal/oxide interface of catalyst nanoparticles. PMID- 16247819 TI - Chain elongation, macrolactonization, and hydrolysis of natural and reduced hexaketide substrates by the picromycin/methymycin polyketide synthase. PMID- 16247820 TI - [Tl8]6- in Cs8Tl8O: a naked eight-vertex closo-deltaeder as a cluster anion. PMID- 16247821 TI - Aluminum-magnesium complexes with linearly bridging carbon dioxide. PMID- 16247822 TI - Pd-catalyzed stereoselective oxidation of methyl groups by inexpensive oxidants under mild conditions: a dual role for carboxylic anhydrides in catalytic C--H bond oxidation. PMID- 16247823 TI - Smooth anodic TiO2 nanotubes. PMID- 16247824 TI - Efficient mono- and bifunctionalization of polyolefin dendrimers by olefin metathesis. PMID- 16247825 TI - On the influence of charged side chains on the folding-unfolding equilibrium of beta-peptides: a molecular dynamics simulation study. AB - The influence of charged side chains on the folding-unfolding equilibrium of beta peptides was investigated by means of molecular dynamics simulations. Four different peptides containing only negatively charged side chains, positively charged side chains, both types of charged side chains (with the ability to form stabilizing salt bridges) or no charged side chains were studied under various conditions (different simulation temperatures, starting structures and solvent environment). The NMR solution structure in methanol of one of the peptides (A) has already been published; the synthesis and NMR analysis of another peptide (B) is described here. The other peptides (C and D) studied herein have hitherto not been synthesized. All four peptides A-D are expected to adopt a left-handed 3(14) helix in solution as well as in the simulations. The resulting ensembles of structures were analyzed in terms of conformational space sampled by the peptides, folding behavior, structural properties such as hydrogen bonding, side chain-side chain and side chain-backbone interactions and in terms of the level of agreement with the NMR data available for two of the peptides. It was found that the presence of charged side chains significantly slows down the folding process in methanol solution due to the stabilization of intermediate conformers with side chain-backbone interactions. In water, where the solvent competes with the solute-solute polar interactions, the folding process to the 3(14)-helix is faster in the simulations. PMID- 16247826 TI - Mechanistic insights into the reversible formation of iodosylarene-iron porphyrin complexes in the reactions of oxoiron(IV) porphyrin pi-cation radicals and iodoarenes: equilibrium, epoxidizing intermediate, and oxygen exchange. AB - We have shown previously that iodosylbenzene-iron(III) porphyrin intermediates (2) are generated in the reactions of oxoiron(IV) porphyrin pi-cation radicals (1) and iodobenzene (PhI), that 1 and 2 are at equilibrium in the presence of PhI, and that the epoxidation of olefins by 2 affords high yields of epoxide products. In the present work, we report detailed mechanistic studies on the nature of the equilibrium between 1 and 2 in the presence of iodoarenes (ArI), the determination of reactive species responsible for olefin epoxidation when two intermediates (i.e., 1 and 2) are present in a reaction solution, and the fast oxygen exchange between 1 and H(2)18O in the presence of ArI. In the first part, we have provided strong evidence that 1 and 2 are indeed at equilibrium and that the equilibrium is controlled by factors such as the electronic nature of iron porphyrins, the electron richness of ArI, and the concentration of ArI. Secondly, we have demonstrated that 1 is the sole active oxidant in olefin epoxidation when 1 and 2 are present concurrently in a reaction solution. Finally, we have shown that the presence of ArI in a reaction solution containing 1 and H(2)18O facilitates the oxygen exchange between the oxo group of 1 and H(2)18O and that the oxygen exchange is markedly influenced by factors such as ArI incubation time, the amounts of ArI and H(2)18O used, and the electronic nature of ArI. The latter results are rationalized by the formation of an undetectable amount of 2 from the reaction of 1 and ArI through equilibrium that leads to a fast oxygen exchange between 2 and H(2)18O. PMID- 16247828 TI - Nucleophilicity-periodic trends and connection to basicity. AB - The potential energy profiles of 18 identity S(N)2 reactions have been estimated by using G2-type quantum-chemical calculations. The reactions are: X- + CH3-X --> X-CH3 + X- and XH + CH3-XH+ --> +HX-CH3 + XH (X = NH2, OH, F, PH2, SH, Cl, AsH2, SeH, Br). Despite the charge difference, the barrier heights and the geometrical requirements upon going from the reactant to the transition structure are surprisingly similar for X- and XH. The barrier heights decrease on going from left to right in the periodic table, and increasing ionization energy (of X- and XH) is correlated with decreasing barrier. The observed trends are explained in terms of substrates with stronger electrostatic character giving rise to lower energetic barriers due to decreased electron repulsion in the transition structure. On the basis of this study, the relationship between the kinetic concept of nucleophilicity and the thermodynamic concept of basicity has been analyzed and clarified. Since the trends in intrinsic nucleophilicity (only defined for identity reactions) and basicity are opposite, overall nucleophilicity (defined for any reaction) will be determined by the relative contribution of the two factors. Only for strongly exothermic reactions will basicity and nucleophilicity be matching. PMID- 16247827 TI - Photophysics of a series of efficient fluorescent pH probes for dual-emission wavelength measurements in aqueous solutions. AB - This paper evaluates the 5-aryl-2-pyridyloxazole backbone to engineer donor acceptor fluorescent pH probes after one- or two-photon absorption. Parent fluorophores, as well as derivatives that can be used to label biomolecules, can be easily obtained in good yields. These molecules exhibit a large one-photon absorption in the near-UV range, and a strong fluorescence emission that covers the whole visible domain. The 5-aryl-2-pyridyloxazole derivatives also possess significant cross sections for two-photon absorption. Upon pyridine protonation, large shifts were observed in the absorption spectra after one- and two-photon excitation, as well as in the emission spectra. This feature was used to measure the pK(a) of the investigated compounds that range between 2 and 8. In most of the investigated derivatives, the pK(a) increased upon light excitation and protonation exchanges took place during the lifetime of the excited state, as shown by phase-modulation fluorometry analysis. Several 5-aryl-2-pyridyloxazole derivatives are suggested as efficient probes to reliably measure the pH of aqueous solutions by means of ratiometric methods that are dependent on fluorescence emission. PMID- 16247829 TI - Homochiral helices of oligonaphthalenes inducing opposite-handed cholesteric phases. AB - The helical structure of the chiral nematic phases (cholesterics) obtained by doping nematic solvents with chiral non-racemic compounds is a macroscopic proof of the solute chirality. Oligonaphthalene (tetra-, hexa-, octa-) derivatives linked at the 1,4-positions have been used as chiral dopants: When the chirality axes are configurationally homogeneous (that is, all-S), the molecular structures correspond to right-handed helices. Yet, we have found series of derivatives with the surprising property that the handedness of the induced cholesteric phase alternates from positive to negative and to positive again, on passing from tetra to hexa- and to octanaphthalene. A comparison with oligonapthalene derivatives, which do not exhibit this twisting ability, points to the importance of the substitution pattern. Both the possibility of inducing oppositely-handed cholesteric phases by homochiral helices of different length, and the role played of substituents, are confirmed by calculations performed with the surface chirality model. PMID- 16247830 TI - Chalcogenide derivatives of imidotin cage complexes. AB - Reaction of the secocubane [Sn3(mu2-NHtBu)2(mu2-NtBu)(mu3-NtBu)] (1) with dibutylmagnesium produces the heterobimetallic cubane [Sn3Mg(mu3-NtBu)4] (4) which forms the monochalcogenide complexes of general formula [ESn3Mg(mu3-NtBu)4] (5a, E = Se; 5b, E = Te) upon reaction with elemental chalcogens in THF. By contrast, the reaction of the anionic lithiated cubane [Sn3Li(mu3-NtBu)4]- with the appropriate quantity of selenium or tellurium leads to the sequential chalcogenation of each of the three Sn(II) centres. Pure samples of the mono- or dichalcogenides are, however, best obtained by stoichiometric redistribution reactions of [Sn3Li(mu3-NtBu)4]- and the trichalcogenides [E3Sn3Li(mu3-NtBu)4]- (E = Se, Te). These reactions are conveniently monitored by using 119Sn NMR spectroscopy. The anion [Sn3Li(mu3-NtBu)4]- also acts as an effective chalcogen transfer reagent in reactions of selenium with the neutral cubane [{Snmu3 N(dipp)}4] (8) (dipp = 2,6-diisopropylphenyl) to give the dimer [(thf)Sn{mu N(dipp)}2Sn(mu-Se)2Sn{mu-N(dipp)}2Sn(thf)] (9), a transformation that results in cleavage of the Sn4N4 cubane into four-membered Sn2N2 rings. The X-ray structures of 4, 5a, 5b, [Sn3Li(thf)(mu3-NtBu)4(mu3-Se)(mu2-Li)(thf)]2 (6a), [TeSn3Li(mu3 NtBu)4][Li(thf)4] (6b), [Te2Sn3Li(mu3-NtBu)4][Li([12]crown-4)2] (7b'') and 9 are presented. The fluxional behaviour of cubic imidotin chalcogenides and the correlation between NMR coupling constants and tin-chalcogen bond lengths are also discussed. PMID- 16247831 TI - Natural products from marine organisms and their associated microbes. AB - The marine environment is distinguished by unique groups of organisms being the source of a wide array of fascinating structures. The enormous biodiversity of marine habitats is mirrored by the molecular diversity of secondary metabolites found in marine animals, plants and microbes. The recognition that many marine invertebrates contain endo- and epibiotic microorganisms and that some invertebrate-derived natural products are structurally related to bacterial metabolites suggests a microbial origin for some of these compounds. Other marine natural products, however, are clearly located in invertebrate tissue and microbial involvement in the biosynthetic process seems unlikely. The complexity of associations in marine organisms, especially in sponges, bryozoans and tunicates, makes it extremely difficult to definitively state the biosynthetic source of many marine natural products or to deduce their ecological significance. Whereas many symbiotic marine microorganisms cannot be isolated and cultured, numerous epi- and endobiotic marine fungi produce novel secondary metabolites in laboratory cultures. The potent biological activity of many marine natural products is of relevance for their ecological function but is also the basis of their biomedical importance. PMID- 16247832 TI - Allosteric control of mRNA decoding. PMID- 16247833 TI - Identification of substrates of the Listeria monocytogenes sortases A and B by a non-gel proteomic analysis. AB - Sortases are enzymes that anchor surface proteins to the cell wall of Gram positive bacteria by cleaving a sorting motif located in the C-terminus of the protein substrate. The best-characterized motif is LPXTG, which is cleaved between the T and G residues. In this study, a non-gel proteomic approach was used to identify surface proteins recognized by the two sortases of Listeria monocytogenes, SrtA and SrtB. Material containing peptidoglycan and strongly associated proteins was purified from sortase-defective mutants, digested with trypsin, and the resulting peptide mixture analysed by two-dimensional nano liquid chromatography coupled to ion-trap mass spectrometry. Unlike enzymes involved in peptidoglycan metabolism, other surface proteins displayed uneven distribution in the mutants. A total of 13 LPXTG-containing proteins were identified exclusively in strains having a functional SrtA. In contrast, two surface proteins, Lmo2185 and Lmo2186, were identified only when SrtB was active. The analysis of the peptides identified in these proteins suggests that SrtB of L. monocytogenes may recognize two different sorting motifs, NXZTN and NPKXZ. Taken together, these data demonstrate that non-gel proteomics is a powerful technique to rapidly identify sortase substrates and to gain insights on potential sorting motifs. PMID- 16247834 TI - Get help for those COPD blues. PMID- 16247835 TI - Chew on this. PMID- 16247836 TI - By the way, doctor. Should I worry if new colon polyps appear where previous ones have been removed? PMID- 16247837 TI - A returning traveler with fever and rash. PMID- 16247838 TI - [New contract recommendations for physicians cooperating with the pharmaceutical industry? The "FS Codex" and its consequences for physicians]. PMID- 16247839 TI - [Research for clinical practice]. PMID- 16247840 TI - The exception to the rule. PMID- 16247842 TI - San Francisco may lower HIV case estimate. PMID- 16247841 TI - Misleading efficacy claims for Fuzeon. PMID- 16247843 TI - Megace suspension approved. PMID- 16247844 TI - Needle-exchange program dashed in Massachusetts. PMID- 16247845 TI - Meth use increases HIV cases in South Dakota. PMID- 16247846 TI - Thoughts catalyzed by Ferro's paper. PMID- 16247847 TI - The door to being preserved and alive. PMID- 16247848 TI - On: Experiences in K and -K. PMID- 16247849 TI - On: The relevance of psychoanalysis to an understanding of terrorism. PMID- 16247850 TI - [Neuroleptic malignant syndrome]. PMID- 16247851 TI - Ischemic preconditioning protects against gap junctional uncoupling in cardiac myofibroblasts. AB - Ischemic preconditioning increases the heart's tolerance to a subsequent longer ischemic period. The purpose of this study was to investigate the role of gap junction communication in simulated preconditioning in cultured neonatal rat cardiac myofibroblasts. Gap junctional intercellular communication was assessed by Lucifer yellow dye transfer. Preconditioning preserved intercellular coupling after prolonged ischemia. An initial reduction in coupling in response to the preconditioning stimulus was also observed. This may protect neighboring cells from damaging substances produced during subsequent regional ischemia in vivo, and may preserve gap junctional communication required for enhanced functional recovery during subsequent reperfusion. PMID- 16247853 TI - Understanding mass panic and other collective responses to threat and disaster. AB - While mass panic (and/or violence) and self-preservation are often assumed to be the natural response to physical danger and perceived entrapment, the literature indicates that expressions of mutual aid are common and often predominate, and collective flight may be so delayed that survival is threatened. In fact, the typical response to a variety of threats and disasters is not to flee but to seek the proximity of familiar persons and places; moreover, separation from attachment figures is a greater stressor than physical danger. Such observations can be explained by an alternative "social attachment" model that recognizes the fundamentally gregarious nature of human beings and the primacy of attachments. In the relatively rare instances where flight occurs, the latter can be understood as one aspect of a more general affiliative response that involves escaping from certain situations and moving toward other situations that are perceived as familiar but which may not necessarily be objectively safe. The occurrence of flight-and-affiliation depends mainly on the social context and especially the whereabouts of familiar persons (i.e., attachment figures); their physical presence has a calming effect and reduces the probability of flight-and affiliation, while their absence has the opposite effect. Combining the factors of perceived physical danger and the location of attachment figures results in a four-fold typology that encompasses a wide spectrum of collective responses to threat and disaster. Implications of the model for predicting community responses to terrorist attacks and/or use of weapons of mass destruction are briefly discussed. PMID- 16247852 TI - Connexin43 PDZ2 binding domain mutants create functional gap junctions and exhibit altered phosphorylation. AB - Connexin43 (Cx43) is the most abundantly expressed gap junction protein. The C terminal tail of Cx43 is important for regulation of gap junctions via phosphorylation of specific tyrosine and serine residues and through interactions with cellular proteins. The C-terminus of Cx43 has been shown to interact with the PDZ2 domain of the tight and adherens junction associated zona occludens 1 (ZO-1) protein. Analysis of the PDZ2 binding domain of Cx43 indicated that positions -3 and -2, and the final hydrophobic amino acid at the C-terminus, are critical for ZO-1 binding. In addition, the C-termini of connexins 40 and 45, but not Cx32, interacted with ZO-1. To evaluate the functional significance of the Cx43-ZO-1 interaction, Cx43 wild type (Cx43wt) and mutants lacking either the C terminal hydrophobic isoleucine (Cx43deltaI382) or the last five amino acids (Cx43delta378-382), required for ZO-1 binding in vitro, were introduced into a Cx43-deficient MDCK cell line. In vitro binding studies and coimmunoprecipitation assays indicated that these Cx43 mutants failed to interact with ZO-1. Confocal and deconvolution microscopy revealed that a fraction of Cx43wt colocalized with ZO-1 at the plasma membrane. A similar colocalization pattern was observed for the Cx43deltaI382 and Cx43 delta378-382 mutants, which were translocated to the plasma membrane and formed functional gap junction channels. The wt and mutant Cx43 appeared to have similar turnover rates. However, the P2 and P3 phosphoisoforms of the Cx43 mutants were significantly reduced compared to Cx43wt. These studies indicated that the interaction of Cx43 with ZO-1 may contribute to the regulation of Cx43 phosphorylation. PMID- 16247854 TI - Crowds and other collectives: complexities of human behaviors in mass emergencies. PMID- 16247855 TI - Emergency evacuations, panic, and social psychology. PMID- 16247856 TI - Wolfgang Amadeus Mozart's psychopathology in light of the current conceptualization of psychiatric disorders. AB - The study of Mozart's letters and biography leads us to reconsider the psychiatric disorders from which he suffered. Indeed, it seems that Mozart demonstrated depressive episodes, some of which were severe and corresponded to the criteria of the DSM-IV classification. However, the arguments put forward by other authors supporting the occurrence of manic or hypomanic episodes (thus constituting a bipolar disorder diagnosis) are not supported by sufficient historic proof. Indeed, the length of time that the behaviors suggesting manic symptoms lasted is not compatible with such a diagnosis. Rather, Mozart's mood swings and impulsive behavior correspond to some traits of a personality disorder, that is, for the most part, symptoms of the dependent personality disorder. Evidence for this diagnosis appears most notably in Mozart's reactions to his wife's absences, but also in occasional behaviors as well as mood lability. The divergences in the classification of Mozart's symptoms, either into the field of bipolar disorders or into that of personality disorders, are closely linked to the nosological uncertainties that are still a source of debate in today's psychiatric research. We discuss a means of overcoming this limitation by considering the concept of "soft bipolar spectrum," a conceptualization that corresponds to Mozart's psychiatric history. PMID- 16247857 TI - Personal narratives of illness in schizophrenia: associations with neurocognition and symptoms. AB - Controversy exists regarding whether unawareness/denial of illness in schizophrenia results from neurocognitive deficits or a rejection of stigmatized social roles. One possibility is that some elements of a narrative of mental illness are primarily a matter of personal/social construction while others may be uniquely curtailed by neurocognitive deficits. Accordingly, we gathered narratives of illness among 52 persons with schizophrenia spectrum disorders using a semi-structured interview. Ratings of the plausibility, adequacy of detail, and temporal conceptual organization of each narrative were correlated with assessments of neurocognition, symptoms, and traditional insight measures. Degree of plausibility was significantly related to performance on the Wisconsin Card Sorting Test (WCST), a measure of executive function and levels of Positive symptoms on the Positive and Negative Syndrome Scale (PANSS). When entered into a regression to predict plausibility, positive symptoms and WCST performance made unique contributions (R2 = .51, p < .0001). Higher levels of Positive symptoms were associated with poorer temporal conceptual organization within narratives. Adequacy of detail within narratives of illness was related to traditional insight measures but not neurocognition or symptoms. PMID- 16247858 TI - A comparison of stigmatizing attitudes toward persons with schizophrenia in four stakeholder groups: perceived likelihood of violence and desire for social distance. AB - This study compared four stakeholder groups regarding the presence of stigmatizing attitudes toward a hypothetical person diagnosed with schizophrenia. Participants included consumers with schizophrenia (n = 104), family members of those with schizophrenia or other severe mental illness (n = 83), mental health clinicians (n = 85), and members of the general public (n = 59); all participants resided in North Carolina. The purpose of the analyses was to investigate whether mental health stakeholder groups differed from the general public and from each other in terms of key attitudes associated with stigmatization of persons with schizophrenia, that is, perceived likelihood of violent behavior, the desire for social distance, and the causes of the illness. Analysis of variance results with follow-up Scheffe tests indicated no statistically significant differences between the mental health stakeholder groups and members of the general public in the likelihood of violence or the desire for social distance. However, there was more variation between both the mental health stakeholder groups and the general public and within the mental health stakeholder groups in the perceptions of the causes of the mental illness. Throughout the analyses, the consumers tended to have the most negative views of the illness. Implications for future stigma research and education are discussed. PMID- 16247859 TI - Differentiating posttraumatic stress between elderly and younger residents. AB - This study aimed to differentiate between the posttraumatic stress responses of elderly and younger community residents who had been exposed to two technological disasters (a train collision and an aircraft crash). One hundred and forty-eight community residents were assessed using the Impact of Event Scale (IES) and the General Health Questionnaire (GHQ-28). The results showed that age differences were not affected by impact of disaster (IES), suggesting that elderly and younger community residents responded to the disasters similarly. Instead, the community residents exposed to the aircraft crash experienced significantly more intrusion and avoidance than those exposed to the train collision. Also, the community residents who had experienced high exposure to the disasters had significantly more intrusive thoughts and exhibited significantly more avoidance behavior than the low/medium exposure group. The results also showed no main effects in general health between the elderly and younger community residents, suggesting that their health status was similar. Instead, the community residents exposed to the aircraft crash had significantly more general health problems than the train disaster residents and the control group. Also, the community residents in either the low/medium or the high exposure group experienced more general health problems than the control group. Correlation coefficients showed that intrusion, avoidance, and the total impact of the disasters were significantly correlated with all general health subscales for both elderly and younger groups. PMID- 16247860 TI - A case of non-psychotic autocastration: the importance of cultural factors. AB - Male genital self-mutilation has been known to occur in psychotic patients. We report a case of premeditated autocastration in the absence of clinical disorder, both at the time of the mutilation and upon follow-up. Coming from a strict Slavic family, this patient repressed gender identity issues for decades. After migrating to Western Europe, his sexual ambiguity became actuated. His motives can be understood in the light of his personal and cultural background. This case shows that cultural factors may contribute to explaining male genital self mutilation behaviors. PMID- 16247862 TI - Quelling the quivers caused by restless leg syndrome. PMID- 16247861 TI - Two case reports of successful prevention of overdosing in borderline patients. AB - Overdosing often occurs among patients with borderline personality disorder (BPD) and confronts physicians with the dilemma of whether to prescribe psychotropic agents for them or not. Prescribing psychotropic agents is often unavoidable in the real world, because psychotherapy alone is sometimes insufficient to control their labile emotions. We report two cases of BPD in which overdosing was successfully prevented by changing the dosage form from tablets to a powder and adding a bitter taste to make it difficult to swallow a large dose at one time. Prevention of overdosing by BPD patients definitely requires a comprehensive treatment approach in terms of its social, biological, psychological, and psychopathological aspects. Although no perfect method of preventing overdosing is currently available, our method provides an option when psychotropic agents are prescribed for BPD patients in clinical practice. PMID- 16247863 TI - What works for difficult-to-treat back pain. PMID- 16247865 TI - Treating a "mini-stroke" to prevent a "major" stroke. PMID- 16247864 TI - First advances in decades in lung cancer care. PMID- 16247866 TI - I have an implantable cardioverter defibrillator (ICD), and I've been taking fish oil supplements because I had heard of their potential heart benefits. However, I recently read they might not be safe for people with ICDs. Should I stop taking them? PMID- 16247868 TI - Benign prostatic hyperplasia: a new look at an old operation. PMID- 16247867 TI - Staying healthy in your car: coping with illness and age. PMID- 16247869 TI - Blood pressure and the heart: lower is better. PMID- 16247870 TI - On call. Your answer to a reader's question in the July issue reassured me (and my wife) that Viagra is safe for my heart. But a new worry has surfaced: is Viagra safe for my eyes? PMID- 16247871 TI - Informational masking release in children and adults. AB - This study assessed informational masking and utilization of cues to reduce that masking in children aged 4-9 years and in adults. The signal was a train of eight consecutive tone bursts, each at 1 kHz and 60 ms in duration. Maskers were comprised of a pair of synchronous tone-burst trains, with randomly chosen frequencies spanning 200-5000 Hz, with a protected region 851-1175 Hz. In the reference condition, maskers were eight bursts in duration, with a fixed frequency within intervals. Experiment 1 tested two monotic masking release conditions: within-interval randomization of masker burst frequency and the introduction of leading masker bursts. Experiment 2 examined masking release in which the signal was presented to one ear and masking components were presented to both ears (masker components in the contralateral ear were 10 dB higher than those in the ipsilateral ear). Both adults and children demonstrated a significant informational masking effect, with children showing a larger effect on average. Both groups also showed significant release from masking in the two monotic conditions, although children received somewhat less benefit from the masking release cues. The binaural condition supported a moderate release from informational masking in adults, but resulted in increased informational masking in children. PMID- 16247872 TI - Preliminary evaluation of prostate cancer metastatic risk biomarkers. AB - Prostate cancer patients at high risk of metastasis need to be identified as early as possible since metastasis is invariably fatal. Treatment could be tailored to risk. Recent array comparative genomic hybridization (aCGH) studies of primary and metastatic prostate tumors identified 39 BAC clones capable of detecting genomic signatures of metastasis. We termed these loci the genomic evaluators of metastatic CaP (GEMCaP). Risk assessments were made on a set of men who were managed with radical prostatectomy. We compared the utility of GEMCaP loci and the Kattan nomogram, a common risk assessment tool, in relation to biochemical outcome. This preliminary evaluation experiment suggests we can use aCGH to detect genomic signatures of metastasis in primary tumors with an accuracy of 78%. The classification accuracy for the Kattan nomogram was 75%. Therefore, validation of GEMCaP is warranted in a larger, appropriately designed cohort. PMID- 16247873 TI - Teaching evidence-based practice on foot. PMID- 16247874 TI - Commentary by Griffiths regarding the Cochrane review on the effectiveness of hip protectors for reducing hip fractures. PMID- 16247876 TI - Review: various devices for delivery of aerosol treatment can be equally efficacious. PMID- 16247875 TI - Clinician advice, an interactive computer program, and motivational counselling during routine medical visits increased reported smoking abstinence among teens. PMID- 16247877 TI - Aspirin, 1000 mg, reduced moderate to severe pain in acute migraine headache. PMID- 16247878 TI - Low dose aspirin lowered stroke risk but not risk of myocardial infarction or cardiovascular death in women. PMID- 16247879 TI - Review: good evidence supports use of polyethylene glycol and tegaserod for constipation. PMID- 16247880 TI - Perceived dynamics of static images enables emotional attribution. AB - Perception of intentions and dispositions of others is an essential ingredient of adaptive daily-life social behaviour. Dynamics of moving images leads to veridical perception of social attributes. Anecdotal observations in art, science, and popular culture indicate that dynamic imbalance can be revealed in static images. Here, we ask whether perceived dynamics of abstract figures is related to emotional attribution. Participants first estimated instability of geometric shapes rotated in 15 degrees steps in the image plane, and then rated the intensity of basic emotions that can be ascribed to the figures. We found no substantial link between the deviation of the figures from the vertical orientation and perceived instability. Irrespective of shape, a strong positive correlation was found between negative emotions and perceived instability. By contrast, positive emotions were inversely linked with deviation of the figure from vertical orientation. The work demonstrates for the first time that dynamics conveyed by static images enables specific emotional attributions, and agrees well with the assumption that neural networks for production of movements and understanding the dispositions of others are intimately linked. The findings are also of importance for exploring the ability to reveal social properties through dynamics in normal and abnormal development, for example in patients with early brain injury or autistic spectrum disorders. PMID- 16247881 TI - Not waving...drowning? PMID- 16247882 TI - Response to Dr Pakula (AFP September). PMID- 16247883 TI - Acupuncture for CME. PMID- 16247884 TI - Caring for carers. PMID- 16247885 TI - Fitness to drive. PMID- 16247886 TI - Saving lives in the aftermath of Pakistan's earthquake. PMID- 16247887 TI - Supply shortages plague Venezuela's public hospitals. PMID- 16247889 TI - Review: non-pharmacological interventions induce or maintain weight loss in adults with prediabetes. PMID- 16247888 TI - [The 49th annual meeting of the Japanese Society for Medical Mycology. Chiba, Japan. October 6-7. 2005. Program and Abstracts]. PMID- 16247890 TI - Physical and functional modularity of the protein network in yeast. AB - While protein-protein interactions have been studied largely as a network graph without physicality, here we analyze two protein complex data sets of Saccharomyces cerevisiae to relate physical and functional modularity to the network topology. We study for the first time the number of different protein complexes as a function of the protein complex size and find that it follows an exponential decay with a characteristic number of about 7. This reflects the dynamics of complex formation and dissociation in the cell. The analysis of the protein usage by complexes shows an extensive sharing of subunits that is due to the particular organization of the proteome into physical complexes and functional modules. This promiscuity accounts for the high clustering in the protein net-work graph. Our results underscore the need to include the information contained in observed protein complexes into protein network analyses. PMID- 16247891 TI - Review: group based education in self management strategies improves outcomes in type 2 diabetes mellitus. PMID- 16247892 TI - Review: self monitoring interventions modestly reduce diastolic blood pressure (BP) but do not improve BP control in hypertension. PMID- 16247894 TI - Review: subglottic secretion drainage reduces ventilator associated pneumonia. PMID- 16247893 TI - Review: some evidence suggests that cognitive behaviour therapy may reduce chest pain in the short term in patients with non-specific chest pain and normal coronary anatomy. PMID- 16247895 TI - [Hasta siempre, commandante!]. PMID- 16247896 TI - Relaxing hip precautions increased patient satisfaction and promoted quicker return to normal activities after total hip arthroplasty. PMID- 16247897 TI - Protein energy supplementation of usual hospital diet did not improve outcomes in inpatients with recent stroke. PMID- 16247898 TI - Timing and route of enteral tube feeding did not affect death or combined death or poor outcome in stroke and dysphagia. PMID- 16247899 TI - Review: early supported discharge reduces death or dependence after stroke. PMID- 16247900 TI - Review: counselling and education may improve outcomes in caregivers of patients with stroke. PMID- 16247901 TI - A multifactorial intervention reduced the mean number of falls but not the proportion who fell in older people with recurrent falls. PMID- 16247902 TI - Review: fever and cough are the most accurate single tests for diagnosing influenza. PMID- 16247903 TI - An educational programme for primary healthcare providers improved functional ability in older people living in the community. PMID- 16247905 TI - Parents of children with diabetes described the transition to the insulin pump in terms of enhanced freedom and quality of life for all family members. PMID- 16247904 TI - Mothers of children at risk described engaging with home visitors in terms of limiting family vulnerability. PMID- 16247906 TI - Parents of young people with mental health problems experienced a deskilling and had to learn to reskill themselves. PMID- 16247907 TI - The media contributed to the knowledge and decision making of patients throughout their cancer experiences. PMID- 16247908 TI - Community living older adults described using medical, collaborative, and self agency models for asthma self management. PMID- 16247909 TI - "Multiple margins" (being older, a woman, or a visible minority) constrained older women's access to Canadian health care. PMID- 16247910 TI - Depressed youth, suicidality and antidepressants. PMID- 16247911 TI - Doing due diligence! Vetting the vendors. PMID- 16247912 TI - Social marketing's unique contributions to health promotion practice. PMID- 16247913 TI - [The evolution of didactic programs of Italian medical faculties towards an European standard]. AB - The article deals with the most recent innovations in the curricula of Italian medical faculties, especially compared with European standards and legislations. PMID- 16247915 TI - Compulsive hoarding: a disease or a sign of a deeper disorder? PMID- 16247914 TI - [Clinical cases and historical-medical didactics]. AB - The discovery of remarkable documents dating back to the thirties and coming from the Policlinico Surgical Clinic of Milan, offers the possibility to be read in a historical clue too and to compare the constant discordances with the clinical contemporary practice. The aim is to point out the relationship between yesterday knowledges and critical today interpretations. The analysis can be suggested because certainly instructive and formative, also within the historical-medical didactics students of the courses of degree in Medicine and Nursing since the first match with the Medical Faculty. PMID- 16247916 TI - [Didactic strategies of Giovan Battista da Monte (1489-1551) for the reformation of medical studies at the Paduan Studium]. AB - Da Monte was a very important member of medical Humanism; his position within the medical cultural scene of his time was characterised by strong interests in the epistemological problems of medicine. He considered the whole medicine - not only theory - a practical science according to the Aristotelian concept of science. During his teaching years at the Paduan Studium he tried to overcome the division of medicine into medical theory and medical practice and he elaborated an unitary method of teaching, trying to follow the Galen's Methodus medendi, in order to give a true scientific foundation also to the medical practice. PMID- 16247917 TI - [The figure of tutor: new perspectives in medical education]. AB - Education in the history of medicine is a crucial issue in constructing a complete medical curriculum. Tutorage in medicine helps the student by building a warm and emotional full relationship between the tutor and the pupil. PMID- 16247918 TI - Moving toward the paperless office. PMID- 16247919 TI - [Ancient instruments, tables, models for a modern education]. AB - The article put a series of actual proposal of scientific education, using historical collections, instruments, tables and models, preserved in Siennese Scientific Museums, as "tools" for the teaching of science. PMID- 16247920 TI - [The historical evolution of obstetrician formation]. AB - The evaluation of the historical development of obstetrician formation presents features of great modernity. At the end of the XVIII century in Florence the first School of Obstetrics for midwives was established. In the School two relevant complementary formative approaches were privileged: ex cathedra didactics and clinical training. The basic tools for these educational methodologies were constituted by handbook production and by wax anatomical models, the latter a field for the meeting and confrontation of surgeons and craftsmen. The historical-social scenario in which this original evolution took place was Tuscany society in the period of Enlightenment, that promoted the formation of professional figures capable of managing natural childbirth in an autonomous way, while nevertheless placing them under a previously lacking health policy control. In this cultural environment particularly impressive was the influence of J.P. Frank (Austria and Lombardy), but also present were other forces, deriving from other European countries, that aimed at including the formation of obstetricians in a scientific programme, thus excluding an unguarded non-structured training. PMID- 16247921 TI - Budgeting gives you a game plan. PMID- 16247922 TI - The stress of an IRS audit can be minimized. PMID- 16247924 TI - Annual ICD-9 update brings new diabetic retinopathy codes. PMID- 16247923 TI - Best clinical practice with ziprasidone IM: update after 2 years of experience. AB - Acute agitation is a common psychiatric emergency often treated with intramuscular (i.m.) medication when rapid control is necessary or the patient refuses to take an oral agent. Conventional i.m. antipsychotics are associated with side effects, particularly movement disorders, that may alarm patients and render them unreceptive to taking these medications again. Ziprasidone (Geodon) is the first second-generation, or atypical, antipsychotic to become available in an i.m. formulation. Ziprasidone IM was approved by the Food and Drug Administration in 2002 for the treatment of agitation in patients with schizophrenia. In October 2004, a roundtable panel of physicians with extensive experience in the management of acutely agitated patients met to review the first 2 years of experience with this agent. This monograph, a product of that meeting, discusses clinical experience to date with ziprasidone IM and offers recommendations on its use in various settings. In clinical trials, patients treated with ziprasidone IM demonstrated significant and rapid (within 15-30 minutes) reduction in agitation and improvement in psychotic symptoms, agitation, and hostility to an extent greater than or equal to that attained with haloperidol i.m. Tolerability of ziprasidone IM was superior to that of haloperidol IM, with a lower burden of movement disorders. Clinical trials have also shown that ziprasidone IM can be administered with benzodiazepines without adverse consequences. Transition from i.m. to oral ziprasidone has been well tolerated, with maintenance of symptom control. The most common adverse events associated with ziprasidone IM were insomnia, headache, and dizziness in fixed dose trials and insomnia and hypertension in flexible-dose trials. No consistent pattern of escalating incidence of adverse events with escalating ziprasidone doses has been observed. Changes in QTc interval associated with ziprasidone at peak serum concentrations are modest and comparable to those seen with haloperidol IM. Results of randomized clinical trials of ziprasidone IM have been corroborated in studies in real-world treatment settings involving patients with extreme agitation or a recent history of alcohol or substance abuse. In these circumstances, clinically significant improvement was seen within 30 minutes of ziprasidone IM administration, without regard to the suspected underlying etiology of agitation. Agents with a good safety/tolerability profile, such as ziprasidone IM, may be more cost effective long term than older agents, due to reduced incidence of acute adverse effects (eg, acute dystonia) that often require extended periods of observation. Additional trials of ziprasidone IM in agitated patients in a variety of clinical setting are warranted to generate comparative risk/benefit data with conventional agents and other second generation antipsychotics. PMID- 16247925 TI - [Ratio compendiaria and visualization of knowledge in anatomical literature (XVI cent.)]. AB - Medical teaching in Modern Age uses anatomical diagrams to implement the discussion on specific topics so to make scientific knowledge comprehensible and remindable. Anatomical diagrams are conceived as reading protocols and as didactic instruments to be used both in a private context of study or during public lessons. PMID- 16247926 TI - [The dialogue between literature and medicine in the education of medical students]. AB - On the basis of the conception that literature, like science, is an instrument of knowledge, the authors used some texts to underline the importance of dialogue between different disciplines. In particular, the discourse on the ethical significance of biomedicine fluctuates from the more established and deep ethics to the humanistic and anthropological ethics present in literature and vice versa. The aim of the work is to demonstrate that literature is an essential study for medical students. Thus, literature does offer the possibility to deepen the awareness of issues and experiences one cannot or will never have. An understanding of narrative can help clinicians and medical students in developing a dynamic relation with everyday complex stories of medical situations. The authors do believe that practising physicians should be encouraged to read critically; every medical and health care students need to be educated how to read in order to learn how to behave in everyday dialogue with patients and their relatives. PMID- 16247927 TI - The responsibilities of tenure evaluators. PMID- 16247928 TI - Hitting the magic angle for cell wall analysis. PMID- 16247929 TI - Analyzing lone lipoproteins. PMID- 16247931 TI - Mars Reconnaissance Orbiter launched. PMID- 16247930 TI - Mapping protein surfaces by MS. PMID- 16247932 TI - Putting a spin on microdevices. PMID- 16247933 TI - Title IX and chemistry. A researcher turns her anger into activism. PMID- 16247934 TI - Organic thin-film transistors as plastic analytical sensors. PMID- 16247935 TI - Rapid LC/MS/MS method development for drug discovery. PMID- 16247936 TI - Seeing (and picking) spots. PMID- 16247937 TI - An epidemiological study of motor neuron disease in Hong Kong. AB - Worldwide, the incidence of motor neuron disease (MND) has been increasing steadily over recent decades. We reported a follow-up epidemiology study of MND in this locality. We identified the subjects from the computer database of the government hospital system between 1 January 1997 and 31 January 2002 by searching the ICD code starting from 335.xx. Every retrieved case or their records were reviewed and validated by neurologist(s) of the responsible regional hospitals which the patients attended. One hundred and twenty cases from seven regional hospitals (serving 48.05% of the HKSAR population) were identified, validated and confirmed to be MND or related diseases. Ninety-eight new cases were diagnosed during the study period. Average age of onset was 58.76 years; SD 14.12 (28-89) years. Male to female ratio was 1.72:1. Peak age of onset was 60-64 years without sex difference. The adjusted incidence rate was 0.60/100,000/year. The adjusted point prevalence at the prevalence date (31 January 2001) was 3.04/100,000. Despite the incidence and prevalence of MND among Hong Kong Chinese, it remained low compared to worldwide figures, and our data suggested a significant rise of MND or related disease in the last decade. A territory-wide prospective epidemiological study is indicated. PMID- 16247938 TI - Metastatic insulinoma and glucagonoma from the pancreas responsible for specific peritumoral patterns of hepatic steatosis secondary to local effects of insulin and glucagon on hepatocytes. PMID- 16247939 TI - Psychological birth of the human grandfather, sometimes a complicated delivery. PMID- 16247940 TI - Practical ethics. Organ donors by Internet. PMID- 16247941 TI - Medication safety issue brief. Counterfeit drug prevention and identification. AB - The U.S. drug supply is purportedly one fo the safest in the world and yet counterfeit medications do make their way to pharmacy shelves and into the hands of unsuspecting patients. The World Health Organization reports that about $35 million in counterfeit drugs are sold n the United States each year. Worldwide, between 6 percent and 10 percent of prescription drugs on the market are counterfeit, WHO estimates. The Food and Drug Administration conducted 22 investigations into counterfeit drugs in 2002 and again in 2003; that's up from an average of five investigations per year through the late 1990s. Counterfeit drugs can cause serious harm to patients, including allergic reactions, and deny them access to potentially life-saving treatments. PMID- 16247942 TI - Many Americans are wary about nursing home care, unclear about paying for it. PMID- 16247943 TI - Screening of newborns prevents brain damage caused by severe jaundice. PMID- 16247944 TI - Autar Singh Paintal (1925-2004). PMID- 16247945 TI - Julius Axelrod (1912-2004). PMID- 16247946 TI - Zebularine: a unique molecule for an epigenetically based strategy in cancer chemotherapy. The magic of its chemistry and biology. AB - 1-(beta-D-ribofuranosyl)-1,2-dihydropyrimidin-2-one (zebularine) is structurally 4-deamino cytidine. The increased electrophilic character of this simple aglycon endows the molecule with unique chemical and biological properties, making zebularine a versatile starting material for the synthesis of complex nucleosides and an effective inhibitor of cytidine deaminase and DNA cytosine methyltransferase. Zebularine is a stable, antitumor agent that preferentially targets cancer cells and shows activity both in vitro and in experimental animals, even after oral administration. PMID- 16247947 TI - Synthesis of complex nucleoside antibiotics. AB - Herbicidin B and fully prtected tunicaminyluracil, which were undecose nucleoside antibiotics, were synthesized using a samarium diiodide (SmI2) mediated aldol reaction with the use of alpha-phenylthioketone as an enolate. The characteristics of the SmI2-mediated aldol reaction are that the enolate can be regioselectively generated and the aldol reaction proceeds under near neutral condition. This reaction is proved to be a powerful reaction for the synthesis of complex nucleoside antibiotics. The synthesis of caprazol, the core structure of caprazamycins, was conducted by the strategy including beta-selective ribosylation without using a neighboring group participation and the construction of a diazepanone by a modified reductive amination. Our synthetic route would provide a range of key analogues with partial structures to define the pharmacophore, which can be a lead for the development of more effective anti bacterial agents. PMID- 16247948 TI - Antiviral potential of a new generation of acyclic nucleoside phosphonates, the 6 [2-(phosphonomethoxy)alkoxy]-2,4-diaminopyrimidines. AB - Three acyclic nucleoside phosphonates (ANPs) have been formally approved for clinical use in the treatment of 1) cytomegalovirus retinitis in AIDS patients (cidofovir, by the intravenous route), 2) chronic hepatitis B virus (HBV) infections (adefovir dipivoxil, by the oral route), and 3) human immunodeficiency virus (HIV) infections (tenofovir disoproxil fumarate, by the oral route). The activity spectrum of cidofovir {(S)- 1-[3-hydroxy-2 (phosphonomethoxy)propyl]cytosine [(S)-HPMPC)]}, like that of (S)-HPMPA [(S)-9-[3 hydroxy-2-(phosphonomethoxy)propyl]adenine) and (S)-HPMPDAP [(S)-9-[3-hydroxy-2 (phosphonomethoxy)propyl]-2, 6-diaminopurine), encompasses a broad spectrum of DNA viruses, including polyoma-, papilloma-, adeno-, herpes-, and poxviruses. Adefovir {9-[2-(phosphonomethoxy)ethyl]adenine (PMEA)} and tenofovir [(R)-9-[2 (phosphonomethoxy) propyl]adenine [(R)-PMPA)]} are particularly active against retroviruses (ie., HIV) and hepadnaviruses (ie., HBV); additionally, PMEA also shows activity against herpes- and poxviruses. We have recently identified a new class of ANPs, namely 6-[2-(phosphonomethoxy)alkoxy]-2,4-diaminopyrimidines, named, in analogy with their alkylpurine counterparts, HPMPO-DAPy, PMEO-DAPy, and (R)-PMPO-DAPy. These compounds exhibit an antiviral activity spectrum and potency that is similar to that of (S)-HPMPDAP, PMEA, and (R)-PMPA, respectively. Thus, PMEO-DAPy and (R)-PMPO-DAPy, akin to PMEA and (R)-PMPA, proved particularly active against HIV- 1, HIV-2, and the murine retrovirus Moloney sarcoma virus (MSV). PMEO-DAPy and (R)-PMPO-DAPy also showed potent activity against both wild type and lamivudine-resistant strains of HBV. HPMPO-DAPy was found to inhibit different poxviruses (ie., vaccinia, cowpox, and orf) at a similar potency as cidofovir. HPMPO-DAPy also proved active against adenoviruses. In vivo, HPMPO DAPy proved equipotent to cidofovir in suppressing vaccinia virus infection (tail lesion formation) in immunocompetent mice and promoting healing of disseminated vaccinia lesions in athymic-nude mice. The 6-[2-(phosphonomethoxy)alkoxy]-2,4 diaminopyrimidines offer substantial potential for the treatment of a broad range of retro-, hepadna-, herpes-, adeno-, and poxvirus infections. PMID- 16247949 TI - A new approach to the synthesis of 4'-carbon-substituted nucleosides: development of a highly active anti-HIV agent 2', 3'-didehydro-3'-deoxy-4'-ethynylthymidine. AB - Oxidation of 3'-O-TBDMS-4',5-unsaturated thymidine 3 with dimethyldioxirane (DMDO) allowed the isolation of the epoxide 4. Upon reacting with organosilicon reagents in the presence of SnCl4, 4 underwent stereoselective ring opening to give 4'-alpha-allyl (6), 4'-alpha-(2-bromoallyl) (7), 4'-alpha-(cyclopenten-3-yl) (8), and 4'-alpha-cyano (9) derivatives of thymidine. Reactions of the 3'-epimer 12 with organoaluminum reagents gave 4'-alpha-methyl (13), 4'-alpha-vinyl (14), and 4'-alpha-ethynyl (15) analogues. Compounds 13-15 were transformed into corresponding 2',3'-didehydro-3'-deoxy derivatives. Evaluation of their ability to inhibit the replication of HIV in cell culture showed that 4'-ethynyl-d4T (19) is more potent and less toxic than the parent compound d4T. PMID- 16247950 TI - Ring-closing metathesis reactions in nucleic acid chemistry-cyclic dinucleotides for targeting secondary nucleic acid structures. AB - Cyclic dinucleotides are synthesized using a ring-closing metathesis protocol and incorporated into oligonucleotides. A stabilization of a three-way junction is observed by an oligodeoxynucleotide containing a central 2'-C to 3'-phosphate connection. PMID- 16247951 TI - Developing synthetic methods for bioactive phosphorus compounds using H phosphonate chemistry: a progress report. AB - In this paper a short account of our recent basic studies aiming toward development of new synthetic methods for the preparation of nucleotide analogues using H-phosphonate chemistry is presented. PMID- 16247952 TI - A nano-circular single-stranded DNA as a novel tool for SNPs detection. AB - Analysis of single nucleotides polymorphisms (SNPs) is very important for the elucidation of gene-based physiological differences. For high-throughput applications, detection systems need to be highly selective, highly sensitive, and simple. In this study, we investigated whether synchronous transcription and translation from nano-circular single-stranded DNA (ssDNA) might be useful for the detection of SNPs. A nano-circular probe ssDNA was designed to contain codons for a (His)6 peptide, and the sense mRNA transcribed from this ssDNA contains a Shine-Dalgarno sequence, a start AUG codon, 6 His codons (CAU), and a stop UAG codon. The entire circular ssDNA is 55 nt and contains T20 as a linker sequence and a binding site for the target SNP-containing DNAs. Our results that show SNPs can be detected by the cell-free synthesis of the (His)6 peptide from this ssDNA. Because this method allows sequence distinction, signal amplification, and easy detection in one system, it should improve the efficacy of high-throughput gene analysis. PMID- 16247953 TI - 6-substituted 5-fluorouracil derivatives as transition state analogue inhibitors of thymidine phosphorylase. AB - A combination of mechanism-based and structure-based design strategies led to the synthesis of a series of 5- and 6-substituted uracil derivatives as potential inhibitors of thymidine phosphorlase/platelet derived endothelial cell growth factor (TP/PD-ECGF). Among those tested, 6-imidazolylmethyl-5-fluorouracil was found to be the most potent inhibitor with a Ki-value of 51 nM, representing a new class of 5-fluoropyrimidines with a novel mechanism of action. PMID- 16247954 TI - Liver targeting of hepatitis-B antiviral lamivudine using the HepDirect prodrug technology. AB - A new class of phosphate and phosphonate prodrugs, called HepDirect prodrugs, has been developed to deliver drugs to the liver while simultaneously diminishing drug exposure to extra-hepatic tissues. The technology combines liver-selective cleavage and kinase by pass with high plasma and tissue stability to achieve increased drug levels in the liver. Lamivudine (LMV), a nucleoside analogue, is a currently approved treatment for hepatitis B infection, but shows modest efficacy and significant drug resistance due to inefficient phosphorylation. LMV is inadequately phosphorylated to the corresponding nucleoside triphosphate in rat and human hepatocytes. A HepDirect prodrug of LMV monophosphate generated 34-fold higher levels of the triphosphate in rat hepatocytes and 320-fold higher triphosphate levels in the liver of treated rats relative to LMV. PMID- 16247955 TI - Synthesis and study of conformationally restricted 3'-deoxy-3',4'-exo-methylene nucleoside analogues. AB - Hitherto unknown restricted 3'-deoxy-3',4'-exo-methylene nucleoside derivatives bearing the nucleic acid naturally occurring pyrimidine bases have been synthesized. The compounds were tested for their activity against HIV, HBV, and several RNA viruses, but they did not show significant antiviral effect. PMID- 16247956 TI - Design and evaluation of 5'-modified nucleoside analogs as prodrugs for an E. coli purine nucleoside phosphorylase mutant. AB - Our studies have led to the identification of an E. coli PNP mutant (M64V) that is able to cleave numerous 5'-modified nucleoside analogs with much greater efficiency than the wild-type enzyme. The biological activity of the three best substrates of this mutant (9-[6-deoxy-alpha-L-talofuranosyl]-6-methylpurine (methyl(talo)-MeP-R), 9-[6-deoxy-alpha-L-talofuranosyl]-2-F-adenine, and 9-[alpha L-lyxofuranosyl]-2-F-adenine) were evaluated so that we can optimally utilize these compounds. Our results indicated that the mechanism of toxicity of methyl(talo)-MeP-R to mice was due to its cleavage to MeP by a bacterial enzyme, and that the toxicity of the two F-Ade analogs was due to their cleavage to F-Ade by mammalian methylthioadenosine phosphorylase. PMID- 16247957 TI - Antiviral activity of steric-block oligonucleotides targeting the HIV-1 trans activation response and packaging signal stem-loop RNAs. AB - Mixmer oligonucleotides consisting of residues of both 2'-O-methylnucleosides (OMe) and locked nucleic acids (LNA) were designed targeting two stem-loops in the 5'-UTR of HIV-1 RNA, the transactivation response region (TAR), which is the site of binding of the Tat protein, and the SL3 loop, which is the primary packaging element that binds the Gag polyprotein. These oligonucleotides were found to inhibit syncitia formation dose- and sequence-dependently when delivered to HeLa T4 LTR beta-Gal cells and subsequently infected with HIV-1. PMID- 16247959 TI - Terminal phosphate labeled nucleotides: synthesis, applications, and linker effect on incorporation by DNA polymerases. AB - A number of terminal phosphate-labeled nucleotides with three or more phosphates and with varied length linkers attached between the terminal phosphate and the dye have been synthesized. These nucleotides have been tested as substrates for different DNA and RNA polymerases. We have also explored their utility in DNA sequencing, SNP analysis, nucleic acid amplification, quantitative PCR, and other biochemical assays. PMID- 16247958 TI - Synthesis and biological evaluation of conformationally restricted and nucleobase modified analogs of the anticancer compound 3'-C-ethynylcytidine (ECyd). AB - A series of conformationally restricted and nucleobase-modified analogs of the anticancer compound 3'-C-ethynylcytidine (ECyd) and its uracil analog (EUrd) have been synthesized. While none of the conformationally restricted analogs displayed anticancer activity, 5-iodo-EUrd and 5-bromo-EUrd displayed potent anticancer activity with IC50 values of 35 nM and 0. 73 microM. PMID- 16247960 TI - Synthesis of novel peptide nucleic acid-peptide chimera for non-invasive imaging of cancer. AB - A chelator-peptide-PNA-peptide chimera specific for KRAS has been prepared by continuous solid phase coupling with a C-terminal insulin-like growth factor 1 (IGF1) ligand, D(cys-ser-lys-cys), and N-terminal bis(S-benzoyl thioglycoloyl) diaminopropanoate chelator for radionuclide labeling. The probe was purified by RP-HPLC and characterized by MALDI-TOF mass spectroscopy. The probe was labeled with 99mTc and 64Cu. Both labeled probes accumulated in human pancreatic cancer xenografts in immunocompromised mice. Control experiments with mismatch chimeras and control xenografts will be necessary to determine the specificity of this molecular diagnostic strategy. PMID- 16247961 TI - Ring opening reactions: synthesis of AICAR analogs as potential antimetabolite agents. AB - In an attempt to improve the AzA selectivity of the 2-(aryl)alkylthio derivatives of adenosine, we planned the synthesis of the corresponding derivatives of the 5 N-ethylcarboxamidoadenosine (NECA). For this purpose, we designed the synthesis of 2-mercapto-NECA to be pursued by means of an opening-closure method We obtained the open AICAR analog; however, ring closure efforts failed to give the desired compound. The newly synthesized AICAR derivative could potentially be endowed with antiviral or antitumoral activity. PMID- 16247963 TI - Synthesis of novel polydiamidopropanoate dendrimer PNA-peptide chimeras for non invasive magnetic resonance imaging of cancer. AB - A variety of dendrimers can be conjugated to oligonucleotides to increase the number of contrast paramagnetic atoms (e.g., gadolinium or dysprosium) per probe. Thus, it was of interest to test a route for assembly of chelating dendrimer branches directly on the N-termini of peptide nucleic acid (PNA)-peptide chimeras by continuous solid-phase coupling on polymer supports. Dendrimer-PNA-peptides complementary to 12 nt of mutant KRAS mRNA have been prepared with a C-terminal insulin-like growth factor 1 (IGF1) analog d(Cys-Ser-Lys-Cys) and N-terminal polydiamidopropanoate (PDAP) dendrimers with different numbers of diaminopropanoate residues. 1,4, 7, 10-Tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) chelating moieties were then coupled to PDAP dendrimer-PNA-peptide chimeras before cleavage from the polymer supports. The DOTA-PDAP-PNA-peptide probes with 1, 2, 4, 8, or 16 amino (or DOTA) moieties were cleaved, purified by RP-HPLC, and characterized by MALDI-TOF mass spectroscopy. PMID- 16247962 TI - Borano-nucleotides: new analogues to circumvent HIV-1 RT-mediated nucleoside drug resistance. AB - Alpha-boranophosphates suppress RT-mediated resistance when the catalytic rate of incorporation (kpol) of the analogue 5'-triphosphate is responsable for drug resistance, such as in the case of K65R mutant and ddNTPs, and Q151M toward AZTTP and ddNTPs. This suppression is also observed with BH3-d4T and BH3-3TC toward their clinically relevant mutants Q151M and M184V. Moreover, the presence of the borano (BH3-) group renders the incorporation of the analogue independent from amino-acid substitutions in RT. To our knowledge, this is the first example of rescue of polymerase activity by means of a nucleotide analogue. PMID- 16247964 TI - Microwave irradiation for accelerating the synthesis of acyclonucleosides of 1,2,4-triazole. AB - The 3-(D-alditol-1-yl)-4-amino-5-mercapto-1,2,4-triazoles 4 and 5 can be successfully prepared using microwave irradiation. Condensation of 4 and 5 with p nitrobenzaldehyde afforded Schiff bases 6 and 7, respectively. Reaction 4 and 5 with ethylchloroacetate gave the corresponding alkylated products 10 and 11. Better yields and much less time were the characteristic features of using the microwave heating over the conventional one. The structure of the prepared compounds was confirmed by 1H-NMR, 2D-NMR and mass spectra. PMID- 16247965 TI - Lentiviral-mediated delivery of combined HIV-1 decoy TAR and Vif siRNA as a single RNA molecule that cleaves to inhibit HIV-1 in transduced cells. AB - RNA interference (RNAi) silences gene expression via short interfering 21-23 mer double-stranded RNA (siRNA) segments that guide cognate mRNA degradation in a sequence-specific manner. On the other hand, HIV-1 decoy TAR RNA are known to competitively interact with the HIV-1 Tat protein, to downregulate the enhanced gene expression from the long terminal repeat (LTR) promoters. Here we report that a novel expression construct, encoding both HIV-1 decoy TAR and Vif siRNA, as a single RNA substrate, was expressed under the control of the human U6 promoter, and later the TAR and siRNA were cleaved into their respective separate RNA by the endogenous RNase III-like enzyme. Each of the cleaved HIV-1 anti-genes then synergistically contributed toward enhancing the inhibition efficacy (>80%) of HIV-1 replication in transduced Jurkat cells. These results suggest that targeting HIV-1 mRNA with simultaneously expressed intracellular decoy TAR and Vif-siRNA could lead to an effective gene therapy strategy for the control and management of HIV-AIDS. PMID- 16247966 TI - E. coli RNase HI and the phosphonate-DNA/RNA hybrid: molecular dynamics simulations. AB - A model for the complex between E. coli RNase HI and the DNA/RNA hybrid (previously refined by molecular dynamics simulations) was used to determine the impact of the internucleotide linkage modifications (either 3-O-CH2-P-O-5' or 3-O P-CH2-O-5) on the ability of the modified-DNA/RNA hybrid to create a complex with the protein. Modified internucleotide linkages were incorporated systematically at different positions close to the 3-end of the DNA strand to interfere with the DNA binding site of RNase H. Altogether, six trajectories were produced (length 1.5ns). Mutual hydrogen bonds connecting both strands of the nucleic acids hybrid, DNA with RNase H, RNA with RNase H, and the scissile bond with the Mg++. 4H2O chelate complex (bound in the active site) were analyzed in detaiL Many residues were involved in binding of the DNA (Arg88, Asn84, Trp85, Trp104, Tyr73, Lys99, Asn100, Thr43, and Asn 16) and RNA (Gln76, Gln72, Tyr73, Lys122, Glu48, Asn44, and Cys13) strand to the substrate-binding site of the RNase H enzyme. The most remarkable disturbance of the hydrogen bonding net was observed for structures with modified internucleotide linkages positioned in a way to interact with the Trp104, Tyr73, Lys99, and Asn100 residues (situated in the middle of the DNA binding site, where a cluster of Trp residues forms a rigid core of the protein structure). PMID- 16247967 TI - A bunch-oligonucleotide forming stable monomolecular quadruplex containing a T tetrad. AB - The chemical synthesis of bunch-ODN I and II prone to form quadruplex structures containing G-and T-tetrads has been reported. Structural studies were performed by 1H-NMR and CD melting experiments. PMID- 16247968 TI - Effects of acrolein on the quadruplex forming d(TTAGGG)4 telomeric repeat sequence. AB - HPLC and ESI-MS analysis have been used to investigate the effect of acrolein exposure on d(TITAGGG)4 human telomeric repeat. Preliminary results disclosed a novel relationship between the structure assumed by oligodeoxynucleotides (ODNs) and the capability of their nucleobase residues to react with acrolein. PMID- 16247969 TI - Phosphate shielding under different conditions results in an enhanced DNA duplex stability. AB - Neutralization of charge of the phosphodiester groups in DNA and the significance for transfer of genetic information will be demonstrated. Theoretical models based on proton shielding are elaborated with ab initio level calculations for a Watson-Crick-type dimer. These results are compared with molecular mechanics studies for duplexes of a hexamer with Rp and Sp phosphate-methylated backbones. PMID- 16247970 TI - The role of the carboxamide group in nicotinamide adenine dinucleotide in relation to hydride transfer: the reduced form of the dinucleotide as remedy in the modulation of neurotransmission. AB - The stereochemical significance of the carboxamide group in combination with the ring nitrogen in NADH-NAD+ conversions has been demonstrated. This has been shown in model systems as well as under enzymatic conditions. The role of the carboxamide group in selective regiospecific interactions has been discussed for neurodegenerative diseases. PMID- 16247971 TI - Spectroscopic and kinetic studies of interactions of calf spleen purine nucleoside phosphorylase with 8-azaguanine, and its 9-(2-phosphonylmethoxyethyl) derivative. AB - Spectroscopic and kinetic studies of interactions of calf spleen purine nucleoside phosphorylase with 8-azaguanine, an excellent fluorescent/fluorogenic substrate for the synthetic pathway of the reaction, and its 9-(2 phosphonylmethoxyethyl) derivative, a bisubstrate analogue inhibitor, were carried out. The goal was to clarify the catalytic mechanism of the enzymatic reaction by identification of ionic/tautomeric forms of these ligands in the complex with PNP. PMID- 16247972 TI - Kinetic model of oxidation catalyzed by xanthine oxidase-the final enzyme in degradation of purine nucleosides and nucleotides. AB - A new kinetic model is presented for analysis of experimental data of oxidation process catalyzed by milk xanthine oxidase. The kinetics for two substrates, xanthine and its analog 2-chloroadenine, in a broad pH range (5.8-9.0) are best described by an equation which is a rational function of degree 2:3 and 2:2, respectively. PMID- 16247973 TI - Kinetic properties of Cellulomonas sp. purine nucleoside phosphorylase with typical and non-typical substrates: implications for the reaction mechanism. AB - Phosphorolysis catalyzed by Cellulomonas sp. PNP with typical nucleoside substrate, inosine (Ino), and non-typical 7-methylguanosine (m7Guo), with either nucleoside or phosphate (Pd) as the varied substrate, kinetics of the reverse synthetic reaction with guanine (Gua) and ribose-1-phosphate (R1P) as the varied substrates, and product inhibition patterns of synthetic and phosphorolytic reaction pathways were studied by steady-state kinetic methods. It is concluded that, like for mammalian trimeric PNP, complex kinetic characteristics observed for Cellulomonas enzyme results from simultaneous occurrence of three phenomena. These are sequential but random, not ordered binding of substrates, tight binding of one substrate purine bases, leading to the circumstances that for such substrates (products) rapid-equilibrium assumptions do not hold, and a dual role of Pi, a substrate, and also a reaction modifier that helps to release a tightly bound purine base. PMID- 16247974 TI - Synthesis and biological evaluation of NAD analogs as human pyridine nucleotide adenylyltransferase inhibitors. AB - NAD analogs modified at the ribose adenylyl moiety, named N-2'-MeAD and Na-2' MeAD, were synthesized as ligands of pyridine nucleotide (NMN/NaMN) adenylyltransferase (NMNAT). Both dinucleotides resulted selective inhibitors against human NMNAT-3 isoenzyme. PMID- 16247975 TI - Design and synthesis of specific inhibitors of the 3'-processing step of HIV-1 integrase. AB - The novel dinucleotide 5'-phosphate, [(L,D)-pIsodApdC], discovered in our laboratory, is a strong inhibitor of HIV-1 integrase for both the 3'-processing and the strand transfer steps. The rationale used in this molecular design was that residues immediately upstream of the dinucleotide cleavage site in the 3' processing step might provide critical recognition/binding sites on integrase. The rationale for the second type of inhibitors was based on the elimination products (linear and cyclic dinucleotides) of 3'-processing. However, while the linear dinucleotide 5'-phosphate (pdGpdT) was active, its cyclic counterpart was inactive against both wild-type and mutant HIV integrase. PMID- 16247976 TI - Naphthyl phosphoramidate derivatives of BVdU as potential anticancer agents: design, synthesis and biological evaluation. AB - The phosphoramidate technology we have developed has been recently applied to BVdU, leading to NB1011 (NewBiotics Inc., California), a novel potential anticancer compound recently entered into phase 2 of the clinical trials for colon cancer. We report in this work a new series of derivatives containing naphthol as aryl masking group on the phosphate moiety, which has shown a significant increase in anticancer activity in preliminary biological evaluations. PMID- 16247977 TI - Control of DNA conformation using 3'-S-phosphorothiolate-modified linkages. AB - An in-depth study into the incorporation of multiple 3-S-phosphorothiolate modifications into oligodeoxynucleotides (ODNs) and their subsequent effect on ODN/DNA and ODN/RNA duplex stability. 3-S-Phosphorothiolate linkages increase the stability of ODN/RNA duplexes and decrease the stability of ODN/DNA duplexes. PMID- 16247978 TI - Targeted therapy of respiratory syncytial virus by 2-5A antisense. AB - Respiratory syncytial virus is a leading cause of respiratory disease in infants, young children, immunocompromized patients, and the elderly. Previous work has shown that RNase L, an antiviral enzyme of the interferon system, can be recruited to cleave RSVgenomic RNA by attaching tetrameric 2' 5'-linked oligoadenylates (2 5A) to an antisense oligonucleotide complementary to repetitive intergenic sequences within the RSV genome (2 5A antisense). RBI034, a 2'-O-methyl RNA-modified analogue of the 2 5A anti-RSV compound, was found to have enhanced antiviral activity in cell culture studies while also cleaving RSV genomic RNA in an RNase L- and sequence-specific manner. RBI034s efficacy in suppressing RSV replication in cell culture is 50 to 100 times better than ribavirin, the only approved drug for RSV infection. Here we show that the activity of 2 SA antisense compound can be further enhanced by a combination treatment with interferon or ribavirin. The anti-RSV activity resulting from combination treatment is more potent than either treatment alone. We also demonstrate that RBI034 is effective against RSV in three different species: mice, cotton rats, and African green monkeys. PMID- 16247979 TI - New affinity resin for purification of cap-binding proteins. AB - Cap binding proteins, which recognize the cap structure present at 5' termini of RNA polymerase II transcripts, have been routinely isolated and purified using affinity resins with mononucleotide cap analogs attached. Here we present a new methodology in which dinucleotide cap analog, m7GpppG, has been linked to the EAH Sepharose. The method is based on derivatization of 2',3'-cis diol of the second nucleotide within the cap structure, with levulinic acid, and subsequent coupling of resulted acetal through its carboxylic group with aminohexyl-agarose. PMID- 16247980 TI - Deaggregation of eIF4E induced by mRNA 5' cap binding. AB - All eukaryotic mRNAs contain a 5' terminal cap structure, which consists of 7 methylguanosine linked by a 5-5' triphosphate bridge to the first transcribed nucleoside (m7GpppN). Specific recognition of the cap by the eukaryotic initiation factor eIF4E plays a key role in regulation of translation initiation as a rate-limiting step. Using dynamic light scattering (DLS), the apo-form of murine eIF4E (33-217) was shown to aggregate. After addition of m7G7P, progressive deaggregation with the time of incubation in the presence of the cap analogue has been observed. PMID- 16247981 TI - Chemical synthesis of the novel Ca2+ messenger NAADP. AB - The first total chemical synthesis of nicotinamide adenine dinucleotide phosphate (beta-NADP, 2) as a single isomer was achieved. This was subsequently converted into the important second messenger nicotinic acid adenine dinucleotide phosphate (p-NAADP) 1 and the identity of this material confirmed by biological evaluation. This flexible synthetic route offers new opportunities for the generation of NAADP 1 analogues that cannot be generated directly from NADP 2 or mainly enzymatic methods. PMID- 16247982 TI - Novel cyclosal nucleotides with reduced inhibitory potency toward human butyrylcholinesterase. AB - Two novel cycloSal-d4T monophosphates (d4TMPs) with increased steric demand have been synthesized via a new synthetic route. While 3-cyclohexyl-cycloSal d4TMP did not show a significantly reduced inhibitory potency toward human butyrylcholinesterase, the opposite was the case for the second novel pronucleotide, bis-(cycloSal-d4TMP). PMID- 16247983 TI - Homo and heterodimers of ddI, d4T and AZT: influence of (5'-5') thiolcabonate carbamate linkage on anti-HIV activity. AB - New homo and heterodimers of ddI, d4T and AZT with (5-5) thiolcarbonate-carbamate linkages have been prepared with the aim of testing them against wild type and NNRTI resistant HIV mutants. The prepared dimers showed a low activity in comparison to the parent drug. PMID- 16247985 TI - Chemical and enzymatic synthesis of 4'-thio-beta-D-arabinofuranosylcytosine monophosphate and triphosphate. AB - N4-Acetyl-1-(2, 3-di-O-acetyl-4-thio-beta-D-arabinofuranosyl) cytosine (2) was synthesized in three steps from 1-(4-thio-beta-D-arabinofuranosyl) cytosine (1). The reaction of this partially blocked 4'-thio-ara-C derivative 2 with 2-chloro 4H-1,3,2-benzodioxaphosphorin-4-one gave the 5-phosphitylate derivative 3, which on reaction with pyrophosphate gave the 5'-nucleosidylcyclotriphosphite 4. Product 4 was then oxidized with iodine/pyridine/water and deblocked with concentrated ammonium hydroxide to provide the desired 4'-thio-ara-C-5' triphosphate 5. This triphosphate 5 was converted to 4'-thio-ara-C -5' monophosphate 6 by treatment with snake venom phosphodiesterase I. The details of the synthesis, purification, and characterization of both nucleotides are described. PMID- 16247984 TI - 3'-modified oligo (2'-O-methylribonucleotides) as improved probes for hybridization with RNA. AB - A series of octa (2-O-methylribonucleotides) with an additional 3'-terminal deoxynucleoside (T, dC, dA or dG) linked by the 3'-3' (inverted) bond was synthesized. The exceptional stability of these oligomers to a 3'-exonuclease (SVP) and nucleases in culture medium containing 10% heat-inactivated fetal calf serum was demonstrated. It was shown that the addition of the 3'-dangling inverted deoxynucleoside increases substantially the thermal stability of the duplexes of oligo(2'-O-methylribonucleotides) with complementary RNA and DNA in the case of a relatively weak terminal AmU(T) pair and enhances the mismatch sensitivity. PMID- 16247986 TI - Synthesis and structural study of quadruplex structures containing 2'-deoxy-8 methyladenosine. AB - The synthesis and preliminary structural studies of ODNs A8MeGGGT and TA8MeGGGT, where A8Me represents 2'-deoxy-8-methyladenosine, are reported. PMID- 16247987 TI - Epoxide adducts at the guanine residue within single-stranded DNA chains: reactivity and stability studies. AB - Emphasis was placed in this work on the assessment of structural and biological features of nucleobase adducts that result from the reaction of DNA with epoxide derivatives. Thus we have prepared and characterized a set of site-specifically modified oligonucleotides at N7-position of a guanine residue, upon reaction with diepoxibutane, with the purpose of further investigating some of their biochemical features. The stability of the lesion-containing DNA fragments has also been investigated and clearly shows that the latter modified oligomers may be used as substrates for in vitro enzymatic assays, aimed at determining the biological effects within cell of these chemically induced DNA damage. PMID- 16247988 TI - Phosphoramidate derivatives of 2',5'-dideoxyadenosine as potential inhibitors of the EDHF phenomenon. AB - P-site inhibitors of adenyl cyclase, such as the dideoxynucleosides 2',3'-ddA and 2',5-ddA, have been shown to attenuate EDHF phenomenon in rabbit arteries and veins. In order to present the dideoxynucleosides as pre-activated nucleotides and bypass the kinase, as well as to prevent their metabolism to dideoxyinosine by adenosine deaminase, the aryloxyphosphoramidate approach has been successfully applied, initially on the 2',3'-ddA. In the present work a new series of 2',5' ddA phosphoramidates has been synthesized, representing the first example of phosphoramidate protide not at the 5'-position. PMID- 16247989 TI - Synthesis and antitumor activity of 5-bromo-1-mesyluracil. AB - Large-scale preparation of 5-bromo-1-mesyluracil (BMsU) 4 has been optimized. BMsU was synthesized by condensation of silylated 5-bromouracil and MsCl in acetonitrile or by the reaction of 5-bromouracil with MsCl in pyridine. The same product was obtained by bromination of 1-mesyluracil. The purpose of this study was to elucidate the effects of BMsU on the biosynthetic activity of tumor cell enzymes involved in DNA, RNA and protein syntheses, and in de novo and salvage pyrimidine and purine syntheses. Investigations were performed in vitro on human cervix carcinoma cells (HeLa). BMsU displayed inhibitory effects on DNA and RNA syntheses in HeLa cells after 24 h of treatment. De nova biosynthesis of pyrimidine and purine was also affected. Antitumor activity of BMsU is closely associated with its inhibitory activity on the enzymes that play an important role in the metabolism of tumor cells. In vivo antitumor activity of BMsU was also investigated. The model used in investigations was a mouse anaplastic mammary carcinoma transplanted into the thigh of the right leg of CBA mice. Significant reduction in tumor growth time was achieved with BmsU administered at a dose of 50 mg/kg. PMID- 16247990 TI - Synthesis and fluorescent properties of the tricyclic analogues of acyclovir linked with nitrogen hetbrocyclic units. AB - Tricyclic (T, 3,9-dihydro-9-oxo-SH-imidazo[g2-c]purine) analogues of acyclovir (ACV, 1), substituted in the 6 position with pyrid-4-yl, 4-(pyrid-4'-yl)Ph, 4 (pyrimidin-5-yl)Ph and 4-(thiazol-2'-yl)Ph units were synthesized. For the synthesis of the heteroarylphenyl derivatives, a convenient general route was developed, ie., Suzuki cross-coupling between protected 6-(4 dihydroxyborylphenyl) TACV and easily available bromoheterocycles. Fluorescent properties of newly synthesized TACV aoalogues strongly depend on the nature of a solvent. This sensitivity of fluorescence makes the compounds promising probes of H-bonding in the environment. PMID- 16247991 TI - Synthesis of 2'-C-methyl-4'-thio ribonucleosides. AB - Starting from 2-C-methyl-ribonolactone, 1,2,3,5-tetra-O-acetyl-2-C-methyl-4 thioribofuranose was synthesized and condensed with heterocyclic bases to afford 2-C-methyl-4'-thioribonucleosides. PMID- 16247992 TI - A direct synthesis of pyrrolocytosine from 5-iodocytosine. AB - We have employed a tandem Sonogashira/annulation reaction between 5-iodocytosine derivatives and terminal alkynes to yield the fluorescent bicyclic nucleobase pyrrolcytosine. Pyrrolocytosine bearing substituents only on the pyrrole ring are conveniently synthesized from 5-iodocytosine. Water soluble pyrrolocytosines are being investigated as reporter groups in SNP analysis. PMID- 16247993 TI - A solid support with a hydroxyallyl linker, full parts of which are potentially reusable for the synthesis of oligonucleotides. AB - This paper describes a solid support with a hydroxyallyl linker that is regenerated without loss of any parts after having used for the synthesis of nucleotides. Reproduction of the solid support can be achieved through detachment of the oligonucleotide by treatment with an organopalladium catalyst in the presence of triethylammonium acetate and subsequent methanolysis of the resulting allyl acetate. PMID- 16247994 TI - Synthesis and enzymatic characterization of methylene analogs of adenosine 5' tetraphosphate (P4A). AB - A new methodology for synthesis of biologically important nucleoside tri- and tetraphosphates containing a bisphosphonate moiety instead of the terminal pyrophosphate bond is described. The series consists of tri- and tetraphosphate analogs of adenosine, guanosine and 7-methylguanosine (characteristic for mRNA cap). We have adopted a two-step procedure that allowed us to insert a methylene bridge into the phosphate chain. Nucleoside mono- or diphosphates were first activated (as imidazole derivatives) and then used in coupling reactions with organic salts of bisphosphonate. The resulting synthetic method enabled us to obtain the desired compounds with high yields and does not require any protective groups. This makes it very useful for the synthesis of labile compounds such as those containing the 7-methylguanosine ring. The structures of the synthesized compounds were confirmed by NMR spectroscopy. They were tested as potential substrates and inhibitors of several hydrolases. PMID- 16247995 TI - Synthesis and properties of mRNA cap analogs containing phosphorothioate moiety in 5',5'-triphosphate chain. AB - Nucleosides and oligonucleotides with an oxygen replaced by sulfur atom are an interesting class of compounds because of their improved stability toward enzymatic cleavage by nucleases. We have synthesized several dinucleotide mRNA cap analogs containing a phosphorothioate moiety in the alpha, beta, or gamma position of 5',5'-triphosphate chain [m7Gp(s)ppG, m7Gpp(s)pG, and m7Gppp(s)G]. These are the first examples of the biologically important 5'mRNA cap analogs containing a phosphorothioate moiety, and these compounds may be useful in a variety of biochemical and biotechnological applications. Incorporation of a sulfur atom in the alpha or gamma position within the dinucleotide cap analog was achieved using PSCl3 in a nucleoside phosphorylation reaction followed by coupling the phosphorothioate of nucleoside with a second nucleotide. Synthesis of cap analogs with the phosphorothioate moiety in beta position was performed using an organic phosphorothioate salt in a coupling reaction with an activated nucleotide. The structures of newly synthesized compounds was confirmed using MS and 1H and 31P NMR spectroscopy. We present here the results of preliminary studies on their interaction with translation initiation factor eIF4E and enzymatic hydrolysis with human and nematode DcpS scavengers. PMID- 16247996 TI - A novel approach to solid phase chemical synthesis of oligonucleotide mRNA cap analogs. AB - A novel approach for the synthesis of 5-capped 2'-O-methyloligoribonucleotides on a disulfide-tethered solid support is described. The key step of the synthesis is ZnCl2 promoted coupling of m7GDP imidazolide to a fully deprotected oligonucleotide 5'-phosphate on-support. By this methodology m7G5'pppm2'Apm2'Upm2'Ap has been prepared. PMID- 16247997 TI - D-4'-thioadenosine derivatives as highly potent and selective agonists at the human A3 adenosine receptor. AB - 4'-Thionucleoside derivatives as potent and selective A3 adenosaine receptor agonists were synthesized, starting from D-gulono-gamma-lactone via D-thioribosyl acetate as a key intermediate, among which the 2-chloro-N6-methyladenosine-5 methyluronamide showed the most potent and selective binding affinity (Ki = 0.28 +/- 0.09 nM) at the human A3 adenosine receptor. PMID- 16247998 TI - Stereoselective synthesis of 3-hydroxymethyl-D-cyclopentenone, the versatile intermediate for the synthesis of carbocyclic nucleosides. AB - The preparative and stereoselective synthesis (45- 50% overall yields, >50 g scale) of the key carbasugars 7a-d was achieved from D-ribose via stereoselective Grignard reaction and oxidative rearrangement as key reactions. PMID- 16247999 TI - Synthesis and biochemical properties of novel mRNA 5' cap analogs resistant to enzymatic hydrolysis. AB - A series of new dinucleotide cap analogs with methylene groups replacing oxygens within the pyrophosphate moieties have been synthesized. All the compounds were resistant to the human scavenger decapping hydrolase, DcpS. Binding constants of the modified caps to eIF4E are comparable to those obtained for m7GpppG. This suggests these methylene modifications in the pyrophosphate chain do not significantly affect cap-binding at least for eIF4E. These cap analogs are also good inhibitors of in vitro translation. mRNAs capped with novel analogs were translated similarly to the mRNA capped with the parent m7GpppG. PMID- 16248000 TI - Microwaves synthesis of solid supports for the synthesis of 3'-aminoalkyl oligodeoxynucleotides. AB - Phthalimido-alkyl alcohol solid supports were rapidly prepared from solid supported phthalic anhydride and amino alcohol condensation induced by microwaves. These supports were used to synthesize 13-aminoalkyl oligodeoxynucleotides allowing a two step deprotection necessary to avoid aminolink alkylation. PMID- 16248001 TI - Novel dinucleoside 5',5'-triphosphate cap analogues. Synthesis and affinity for murine translation factor eIF4E. AB - Chemical synthesis of a series of novel dinucleoside cap analogues, m7GpppN, where N is formycin A, 3'-O-methylguanosine, 9-beta-D-arabinofuranosyladenine, and isoguanosine, has been performed using our new methodology. The key reactions of pyrophosphate bonds formation were achieved in anhydrous dimethylformamide solutions employing the catalytic properties of zinc salts. Structures of the new cap analogues were confirmed by 1H NMR and 31p NMR spectra. The binding affinity of the new cap analogues for murine eIF4E(28-217) were determined spectroscopically showing the highest association constant for the analogue that contains formycin A. PMID- 16248002 TI - Combination of poly(L-lysine)-graft-dextran copolymer and 2'-O,4'-C-methylene bridged nucleic acid (2',4'-BNA) modification synergistically stabilizes pyrimidine motif triplex at neutral pH. PMID- 16248003 TI - Synthesis and biological evaluation of N- and O-alkylated bicyclic furanopyrimidines as non-nucleosidic inhibitors of human cytomegalovirus. AB - 2'3'-Dideoxy furanopyrimidines were shown to display anti-HCMV activity via a non nucleoside mechanism. Further studies into highly modified sugar derivatives led to the preparation of N-and O-alkylated C10 furanopyrimidine analogues, and this work is described herein. These compounds were tested against HCMV strains, and the first case of submicromolar activity was observed. PMID- 16248004 TI - The journey towards elucidating the anti-HCMV activity of alkylated bicyclic furano pyrimidines. AB - Bicyclic furanopyrimidines were recently discovered by us to be potent and selective inhibitors of VZV. Related studies to investigate the role of the sugar in this activity uncovered dideoxy furanopyrimidines as inhibitors of HCMV and this led to the preparation of highly modified long alkyl chain furanopyrimidines from the N- and O-alkylation of their parent bases. Herein we describe their synthesis and subsequent biological evaluation against HCMV. O-alkylated derivatives were almost invariably found to be at least equiactive with their N alkylated counterparts. At this point, little change in activity has been found with large variation in N- and O-substituent. PMID- 16248005 TI - Anti-tumor effect of intravenous TNFalpha gene delivery naked plasmid DNA using a hydrodynamics-based procedure. AB - High levels of foreign gene expression in mouse hepatocytes can be achieved by the rapid injection of a large volume of naked plasmid (pDNA) into animals via the tail vein, the so-called hydrodynamics-based procedure. In this study, we evaluated the efficacy of hydrodynamics-based tumor necrosis factor alpha (TNFalpha) transfer for tumor treatment, in which the naked pDNA encoding TNFalpha was administered into the tail vein following an intravenous injection of B16 melanoma cells. The mice treated with TNFalpha-expressing pDNA displayed a profound reduction in lung metastasis. These results suggest that the hydrodynamics-based transfer of naked pDNA is a convenient and efficient method of TNFalpha gene therapy against metastatic tumors. PMID- 16248006 TI - Synthesis and purification of fluorinated benzimidazole and benzene nucleoside-5' triphosphates. AB - The expression "universal base" is very often used to express hybridization properties and recognition patterns of nucleosides. Their behaviour in biological applications, however, is of great interest regarding, e.g.,' their incorporation by polymerases. The 4,6-difluorobenzimidazole and the 2,4-difluorobenzene nucleoside analogues have proven to be universal bases that do not discriminate between the four natural nucleobases in RNA duplexes. Therefore, we synthesized the corresponding triphosphates to evaluate their behavior in polymerase catalyzed reactions and to investigate their ability to serve as substrates for the T7 RNA polymerase. PMID- 16248007 TI - Synthesis and biological activity of cyclic ADP-carbocyclic-ribose analogs: structure-activity relationship and conformational analysis of N-1-carbocyclic ribose moiety. AB - Several cyclic ADP-carbocyclic-ribose analogs 3-10 modified in the N-1 carbocyclic-ribose moiety were synthesized. Their Ca2+-releasing activity was estimated in sea urchin eggs to show that the 3"-deoxy analog 6 shows 5 times more potent activity than cADPcR, but the 2",3"-didieoxy-2",3"-unsunsaturated analog 3 has very weak activity. We also calculated their stable conformation and found that 3 and 6 were significantly different in their stable conformation. PMID- 16248008 TI - Oxidative transformations of nucleoside fluorenemethyl H-phosphonoselenoate diesters. AB - 9-Fluorenemethyl H-phosphonoselenoate monoester has been used to produce thymidine 3'-O-phosphoroselenoate monoester from which various P(V) derivatives containing multiple modifications at phosphorus were obtained; e.g., thymidine 3' O-phosphoroselenofluoridate, 3'-O-phosphoroselenothioate, or 3'-O phosphorodiselenoate monoesters. PMID- 16248009 TI - Accurate mass analysis of phosphoramidites by electrospray mass spectrometry. AB - A method of accurate mass determination of phosphoramidites is described. The commonly used methanol/water/acid system was replaced by LiCl-containing acetonitrile and the concentrations of LiCl, poly(ethylene glycol), and phosphoramidite samples were optimized. PMID- 16248010 TI - Synthesis of 2'-C-methyl-beta-D-ribofuranosylimidazo [4,5-d]-pyridazine derivatives (2-aza-3-deazapurine nucleoside analogues). AB - In order to evaluate their antiviral properties, two imidazo[4,5-d]pyridazine 2' C-methyl-beta-D-ribofuranonucleoside derivatives have been synthesized. PMID- 16248011 TI - Synthesis of 5-aza-7-deazaguanine nucleoside derivatives as potential anti flavivirus agents. AB - Coupling suitable sugars (D- or L-ribofuranose, 2' or 3-deoxysugar, branched sugars) with 2-aminoimidazo[1,2-a]-s-triazin-4-one was carried out using the different reaction conditions: 1) condensation in the presence of sodium hydride; or 2) condensation using Vorbruggen's methods. The 5-aza- 7-deazaguanine nucleoside analogues obtained were evaluated in cell culture experiments for the inhibition of the replication of a number of RNA viruses, including BVDV, YFV, and WNV. PMID- 16248012 TI - Model synthesis of nucleoside boranophosphoramidate with amino acid for prodrug purpose. AB - A model synthesis of a nucleoside boranophosphoramidate prodrug with (L) tryptophan methyl ester was accomplished in a one-pot reaction via an H phosphonate approach. This new type of compound is expected to possess the potent antiviral and anticancer advantages conferred by boranophosphates and normal nucleoside amino acid phosphoramidate. PMID- 16248013 TI - DNA self-assembling systems: branched oligonucleotides as building blocks and monitoring by pyrene excimer band formation. AB - The construction of a DNA self-assembling system created by four Y-shaped branched oligodeoxynucleotide building blocks has been studied. The assembly was verified by changes in the fluorescence emission spectra and revealed an additive effect in pyrene excimer band formation during DNA self-assembly. PMID- 16248014 TI - Synthesis of carbocyclic nucleotides as potential substrates for thymidylate kinase. AB - Enantiomerically pure carbocyclic 2'-deoxy-3'-azidothymidine monophosphate (AZTMP) and carba-2'deoxy-3'-thiocyanatothymidine monophosphate were synthesized to study their behavior toward their phosphorylation by thymidylate kinase. The nucleotides were synthesized starting from the parent nucleosides by an alkaline hydrolysis of the corresponding cycloSal-phosphate triesters. PMID- 16248015 TI - Engineering DNA topology with locked nucleosides: a structural study. AB - DNA dodecamers modified with nucleotide building blocks based on a bicyclo[3. 1.0]hexane system that effectively locks the ribose template into an RNA-like or North (N) conformation were analyzed by various biophysical techniques including high field nuclear magnetic resonance (NMR). Replacement of either one or both of the center thymidines in the Dickerson Drew dodecamer (CGCGAAT*T*CGCG) caused a progressive shift in the bending propensity of the double helix as shown by a newly developed rapid technique that compares the residual dipolar coupling (RDC) values of the modified duplexes with those previously determined for the native DNA. PMID- 16248016 TI - Synthesis of oligonucleotides bearing an arylamine modification in the C8 position of 2'-deoxyguanosine. AB - C8-Arylamine-dG adducts were converted into their corresponding 5-O-DMTr-3'-O phosphoramidite-C8-arylamine-dG derivatives. These compounds were used for the automated synthesis of site-specifically modified oligonucleotides. The oligonucleotides were studied for their CD properties, Tm values, and their effects on primer extension assays using human DNA-polymerase beta. PMID- 16248017 TI - Rapid access to 2'-branched-carbocyclic nucleosides and their 4'-epimers from 2 alkyl-cyclopentene-1-ones. AB - 2-Methyl-2-cyclopentene-1-one was used as starting material in a novel route toward 2'-branchedcarbocyclic nucleosides. This methodology was efficiently adapted to the preparation of 4'-epicarbocycles. A series of this new class of molecules was synthesized as potential antiviral compounds. PMID- 16248018 TI - Comparative conformational analysis of nucleosides by NMR, X-ray, and semi empirical (PM3 VS. AM1) methods. AB - The 2'-deoxy-2'-fluoro-beta-D-arabinofuranosyl ortho-aza-purine and -pyrimidine nucleosides manifest an unusually rigid sugar N conformation in solution. PMID- 16248019 TI - Structure-activity relationship of 5'-substituted fluoro-neplanocin a analogues as potent inhibitors of S-adenosylhomocysteine hydrolase. AB - Four 5'-substituted fluoro-neplanocin A analogues la-d were designed and synthesized, and the inhibitory activity against SAH was in the following order: NH2 > SH > F, N3, indicating a hydrogen bonding donor is essential for inhibitory activity. PMID- 16248020 TI - Stereoselective synthesis of conformationally rigid apio carbanucleosides as potential antiviral agents. AB - Apio north-methanocarbocyclic nucleosides 1-3 with bicyclo[3. 1.0]hexane template were first synthesized. Introduction of hydroxymethyl substituent was efficiently and stereoselectively accomplished by aldol and retro-aldol reaction and fixed conformation was achieved from a modified Simmons-Smith cyclopropanation on a cyclopentane ring. PMID- 16248021 TI - Synthesis of some 2'-fluoro-2'-deoxy-3'-C-ethynyl and 3'-C-vinyl-beta-D lyxofuranosyl nucleosides. AB - 1-(2-Fluoro-2-deoxy-beta-D-drabinofuranosyl) uracil (5) and 1-(2-fluoro-2-deoxy beta-D-arabinofuranosyl)cytosine (6) were synthesized as reported earlier. Both of these compounds were converted into 2'-fluoro-2'-deoxy-3'-C-ethynyl and 3'-C vinyl-beta-D-lyxofuranosyl nuclearsides (16-19) by a multistep sequence. All these new nucleosides were evaluated against seven human tumor cell lines in vitro. PMID- 16248022 TI - Inhibitors of inosine monophosphate dehydrogenase: probes for antiviral drug discovery. AB - The role of inosine monophosphate dehydrogenase (IMPDH) at the metabolic branch point of de novo purine nucleotide biosynthesis makes this enzyme an attractive probe for the discovery of antiviral compounds. Introduction of unsaturation at the 2-position of IMP, the natural substrate for IMPDH, produces Michael acceptors at that position, which results in these compounds being inhibitors of IMPDH. Consistent with this mechanism-based molecular design, some of the parent nucleosides exhibited antiviral activity. PMID- 16248023 TI - Design of highly efficient and selective transfer reaction of nitrosyl group to dc and d(M)C resulting in specific deamination. AB - Nitric oxide (NO) is an important endogenous regulatory molecule, and S nitrosothiols are believed to play a significant role in NO storage, transport, and delivery. Based on the ability to generate NO in vivo, S-nitrosothiols can be used as therapeutic drugs. In this study, we have developed an innovative method for sequence- and base-specific delivery of NO to a specific site of DNA followed by specific deamination. PMID- 16248024 TI - Ring-expanded analogues of natural oxetanocin: (+) and (-) hydroxymethyl isodideoxyadenosine. AB - New enantiomeric isonucleoside analogues related to natural oxetanocin have been synthesized from D-glucosamine and D-glucose. The structures of the target compounds were confirmed by NMR, HRMS, UV, single crystal X-ray, and optical rotation data. Stability studies with respect to purine nucleoside phosphorylase and adenosine deaminase show that these compounds are not substrates. Antiviral results are discussed. PMID- 16248025 TI - 2'-bis-pyrene modified oligonucleotides: sensitive fluorescent probes of nucleic acids structure. AB - A new type of fluorescent nucleic acid probes, 2-bis-pyrene-modified oligonucleotides, is described. Preparation of these conjugates involves attachment of two pyrene moieties to the 2'-phosphate group introduced into any position within a sequence by solid-phase phosphoramidite synthesis. Good hybridization properties of the 2'-bis-pyrene probes, their nuclease resistance and sensitivity of fluorescence to the type of complementary nucleic acid have been demonstrated. PMID- 16248026 TI - Synthesis of a new N-9 ribityl analogue of cyclic inosine diphosphate ribose (cIDPR) as a mimic of cyclic ADP ribose (cADPR). AB - A new analogue of cyclic inosine diphosphate ribose (cIDPR), in which the N-1 and N-9 ribosyl moieties were substituted by a carbocyclic moiety and a hydroxyl alkyl chain, has been synthesized and characterized. PMID- 16248027 TI - Unusual monomolecular DNA quadruplex structures using bunch-oligonucleotides. AB - The chemical synthesis of several G-rich bunch-oligonucleotides and the structural characterization of the corresponding monomolecular G-quadruplexes (I IV) have been reported. The synthetic method allow the achievement of monomolecular DNA quadruplex structures having unusual and predeterminable oligodeoxyribonucleotide (ODN) strand orientation. PMID- 16248028 TI - An intramolecular 1,3-dipolar cycloaddition reaction towards the synthesis of chiral azetidine nucleoside analogues: the D-gluco case. AB - The diastereoselective intramolecular 1,3-dipolar cycloaddition reaction of unsaturated nitrones, derived from methyl alpha-D-glucopyranoside with 2 furaldehyde and 2-(benzyloxy)acetaldehyde has been studied In our pevious studies with 2-furaldehyde, the cycloaddition resulted 3 diastereoisomers in a 3:1:1 ratio. In this article, how the number of the possible isomers generated by 1,3 cycloaddition could be reduced from 4 to 1 when 2-(benzyloxy)acetaldehyde was employed as an aldehyde is shown. PMID- 16248029 TI - Mild, efficient, selective and "green" benzoylation of nucleosides using benzoyl cyanide in ionic liquid. AB - Use of benzoyl cyanide (BzCN) for benzoylation of nucleosides has been studied, both in pyridine and in ionic liquid BzCN in 1-methoxyethyl-3-methylimidazolium methanesulfonate as ionic liquid has been found to be a "green "alternative compared to the pyridine-BzCN system. An efficient and selective benzoylation of nucleosides of both, the 2'-deoxy- and the ribo-series at ambient temperature was accomplished. PMID- 16248030 TI - Interaction of porphyrin with G-quadruplex structures. AB - Isothermal titration calorimetry (ITC) is a sensitive technique for probing bimolecular processes and can provide direct information about the binding affinity and stoichiometry and the key thermodynamic parameters involved. ITC has been used to investigate the interaction of the ligand H2TMPyP to the two DNA quadruplexes, [d(AGGGT)]4 and [d(TGGGGT)]. Analysis of the ITC data reveals that porphyrin/quadruplex binding stoichiometry under saturating conditions is 1:2 for [d(AGGGT)]4 and 2:1 for [d(TGGGGT)], respectively. PMID- 16248031 TI - Relative stability of quadruplexes containing different number of G-tetrads. AB - The aim of this work is to compare the physicochemical properties of three oligonucleotidic sequences, d(TGGGT), d(TGGGGT) and d(TGGGGGT), which assemble to form quadruplex structures with the same molecularity, but containing three, four, and five G-quartets, respectively. The addition of one or two G-tetrads greatly increases both the enthalpy and Tm values of the quadruplex dissociation. PMID- 16248032 TI - The synthesis of diene-containing nucleoside phosphoramidites and their use in the labeling of oligonucleotides. AB - A variety of furan-modified nucleoside phosphoramidite monomers has been prepared and efficiently incorporated into oligonucleotides. These take part in Diels Alder reactions with fluorescent maleimides to give fluorescent-labeled oligonucleotides. This represents a strategy for oligonucleotide labeling that is orthogonal to amine-based methods. PMID- 16248033 TI - Nm 283, an efficient prodrug of the potent anti-HCV agent 2'-C-methylcytidine. AB - In order to improve the oral bioavailability of 2-C-methylcytidine, a potent anti HCV agent, the corresponding 3'-O-L-valinyl ester derivative (NM 283) has been synthesized Based on its ease of synthesis and its physicochemical properties, NM 283 has emerged as a promising antiviral drug for treatment of chronic HCV infection. PMID- 16248034 TI - Synthesis of spin-labeled RNAs for long range distance measurements by peldor. AB - Site directed spin labeled RNA duplexes with different interspin distances were synthesized. The radical 2,2,5,5-tetramethyl-pyrrolin- 1-yloxyl-3-acetylene (TPA) was introduced during the solid-phase synthesis through a Sonogashira cross coupling with 5-iodo-uridine. Tm and CD studies showed that the spin label does not to disturb significantly the A-form of these duplexes. 4-Pulse Electron Double Resonance (PELDOR) was then used to measure intramolecular spin-spin distances of 19.3, 33.0 and 40.9 A, which are in very good agreement with the calculated values of 17.6, 32.1 and 39.1 A, obtained from Molecular Dynamics (MD) simulations. PMID- 16248035 TI - Large-scale synthesis of "Cpep" RNA monomers and their application in automated RNA synthesis. AB - Small interfering RNAs (siRNA) are the latest candidates for oligonucleotide based therapeutics. Should siRNA be successful in clinical trials, a huge demand for synthetic RNA is anticipated. We believe that 1-(4-chlorophenyl)-4 ethoxypiperidin-4-yl (Cpep) is an ideal 2'-protecting group for large-scale syntheses. Unlike 2'-silyl groups, mild acid hydrolysis instead of fluoride ion is used for the 2'-deprotection. The syntheses of 2'-Cpep protected nucleosides (A, C, G, and U) has been accomplished on a 0.5 Kg scale. The 2'-Cpep monomers were transformed into 3'-O-phosphoramidites for conventional automated solid phase synthesis. Cost-effective processes for large-scale synthesis of Cpep monomers and initial automated solid-phase synthesis are demonstrated. PMID- 16248036 TI - Effects of a 8-oxoadenosine incorporation on quadruplex structures: thermal stabilities and structural studies. AB - The effects of incorporation of 8-oxoadenosine in two different truncations of human telomeric sequence forming quadruplex structures are reported. In order to characterise their structures, a combination of NMR and UV spectroscopy and computational techniques were used. Both oligonucleotides have been found to form fourfold symmetric quadruplex structures. As a tautomeric equilibrium between keto and enol forms of 8-oxoadenosine may establish in solution and intrinsic stabilities effects, such as internal H-bonds, for example, may determine the predominance of some particular tautomer, molecular modelling studies were performed on quadruplex structures containing both the tautomeric forms. Both molecules resulted to be thermally less stable than the natural. PMID- 16248037 TI - Molecular modeling studies of a parallel stranded quadruplexes containing a 8 bromoadenosine. AB - Truncated sequences of human telomeric DNA can readily assemble to form parallel stranded quadruplexes containing A- and G-tetrads. The formation of an A-tetrad is highly context-dependent and the relationship between the formation of an A tetrad and the glycosidic torsion angle of the adenosine residues implicated has not been completely clarified so far. In order to give a further insight in this issue we synthesized the modified oligomers d(ABrGGGT) and d(TABrGGGT), two different truncations of the human telomeric sequence containing a 8 bromoadenosine residue, named ABr. NMR data show that both the modified oligomers are able to perfectly fold into highly symmetric quadruplexes with all strands parallel to each other. Molecular modeling studies were performed on both [d(ABrGGGT)]4 and [d(TABrGGGT)]4, indicating that a bulky substituent, such as a bromine atom at the C8 position of adenines, can force the glycosidic bond to adopt a syn conformation, stabilizing the resulting quadruplexes. PMID- 16248038 TI - Structural studies on LNA quadruplexes. AB - LNAs (locked nucleic acids) are new DNA analogues with higher binding affinities toward nucleic acids than the canonical counterparts mainly due to the characteristic conformational restriction arising from the 2'-O, 4'-C methylene bridge. In light of the promising therapeutic applications and considering the advantageous characteristics of LNAs, such as their high water solubility, easy handling, and synthetic accessibility through the conventional phosphoramidite chemistry, we undertook a study concerning the capability of these nucleic acid analogues to form quadruplex structures. Particularly, we have been investigating the LNA/DNA chimeras corresponding to the well-known DNA sequences 5 GGTTGGTGTGGTTGG-3', capable of forming an unimolecular quadruplex. This article deals with the study of the sequence 5'-ggTTggTGTggTTgg-3' (upper and lower case letters represent DNA and LNA residues, respectively), which, according to CD spectroscopy, is able to fold into a quadruplex structure. PMID- 16248039 TI - Synthesis of novel piperidinyl linker based energy transfer terminators and their potential use in DNA sequencing. AB - Synthesis of novel piperidinyl linker based ET cassettes and terminators is described These novel terminators are evaluated in the DNA sequencing experiments using thermostable DNA polymerase. PMID- 16248040 TI - Synthesis of racemic and enantiomeric 3-pyrrolidinyl derivatives of purine and pyrimidine nucleobases. AB - The present work relates to the synthesis of pyrrolidine nucleoside analogs. Starting from malic acid, we have elaborated a high-yield synthesis of racemic and enantiomeric N-protected 3-pyrrolidinols and their O-mesyl derivatives as key compounds for alkylations of purine and pyrimidine nucleobases. On varying base and solvent, we have found conditions providing both satisfactory N-/O regioisomeric ratio and acceptable yield for pyrimidine compounds. PMID- 16248042 TI - Kilo-scale synthesis process for 2'-O-(2-methoxyethyl)-pyrimidine derivatives. AB - We describe an improved process to produce 2'-O-(2-methoxyethyl)-pyrimidines. Starting with commercially available O-2,2'-anhydro-5-methyluridine and tris-(2 methoxyethyl)borate, we modified the ring-opening reaction conditions and changed to a continuous extraction purification method to give 2'-O-(2-methaxyethyl)-5 methyluridine. The dimethoxytritylation 5'/3' ratios and yield were improved by the use of 2,6-lutidine as the base. Conditions to convert to the 5' methylcytidine analog and its isolation by crystallization were optimized. Final benzoylation was improved by developing a method to selectively hydrolyze benzoyl ester impurities. PMID- 16248041 TI - Bis(tBuSATE) phosphotriester prodrugs of 8-azaguanosine and 6-methylpurine riboside; bis(pom) phosphotriester prodrugs of 2'-deoxy-4'-thioadenosine and its corresponding 9alpha anomer. AB - As an extension of previous work with bis(POM) nucleotide prodrugs, we report the synthesis and biological evaluation in tumor cell culture of the bis(pivaloyloxymethyl) phosphotriester prodrug of slightly cytotoxic 2'-deoxy-4' thioadenosine and its alpha-anomer. We have experienced need for an alternative phosphate masking group, particularly with purine nucleosides. Accordingly, we report synthesis and biological evaluation of the bis(tBuSA TE) phosphotriester prodrugs of 8-azaguanosine and 6-methylpurine riboside, nucleoside analogs with moderate to significant cytotoxicity. All four prodrugs were examined in tumor cell culture in parallel with the parent nucleosides. Synthetic routes and biological data are presented. PMID- 16248043 TI - Transprotection of silyl ethers of nucleosides in FeCl3 based ionic liquids. AB - Ionic liquid mediated deprotection of tert-butyldimethyl silyl (TBDMS) ethers derived from various primary and secondary alcohols have been studied and the reaction conditions optimized. Deprotection of the silyl ethers in FeCl3 based ionic liquids in presence of acetic anhydride yielded the acetate esters of the corresponding alcohols in good yields. The transprotection methodology was extended to the silyl ethers of nucleosides to yield the corresponding acetylated products. PMID- 16248044 TI - Cdk inhibitory nucleoside analogs prevent transcription from viral genomes. AB - Targeting viral proteins has lead to many successful antivirals. Yet, such antivirals rapidly select for resistance, tend to be active against only a few related viruses, and require previous characterization of the target proteins. Alternatively, antivirals may be targeted to cellular proteins. Replication of many viruses requires cellular CDKs and pharmacological CDK inhibitors (PCIs), such as the purine-based roscovitine (Rosco), are proving safe in clinical trials against cancer. Rosco inhibits replication of wild-type or (multi-)drug resistant HIV, HCMV, EBV, VZV, and HSV-1 and 2. However, the antiviral mechanisms of purine PCIs remain unknown. Our objective is to characterize these mechanisms using HSV as a model We have shown that Rosco prevents initiation of transcription from viral, but not cellular, genomes. This inhibition is promoter independent, but genome dependent, and requires no viral proteins. This is a novel antiviral mechanism and a previously unknown activity for purine PCIs. PMID- 16248045 TI - 7-deazapurin-2,6-diamine and 7-deazaguanine: synthesis and property of 7 substituted nucleosides and oligonucleotides. AB - The synthesis of 7-substituted 7-deazaguanine and 7-deazaadenine ribonucleosides 1-2, the incorporation of 3a-d into oligonucleotides, and the stability of the corresponding duplexes and base discrimination are described. The pKa values of 3 4 are determined. PMID- 16248046 TI - Gold DNA-conjugates: ion specific self-assembly of gold nanoparticles via the dG quartet. AB - The Oxytricha telomere DNA hairpin 5'-d(G4T4G4) immobilized on 13 nm gold nanoparticles forms a supramolecular assembly via dGC-quartets, as determined by the color change and by SEM. The aggregation is ion-dependent and selective for sodium ions. K+ is less efficient while Li+ and Cs+ do not drive the aggregation. This work is the first effort exploring the use of secondary structures of DNA (quadruplexes) for producing self-assemblies of gold nanoparticles. PMID- 16248047 TI - 6-azapyrimidine and 7-deazapurine 2'-deoxy-2'-fluoroarabinonucleosides: synthesis, conformation and properties of oligonucleotides. AB - The synthesis of 2'-deoxy-2'-fluoro-beta-D-arabinofuranosyl nucleosides (1b, 2b, and 3b) were described and their conformation in solution as well as in the solid state was determined In addition to this, building blocks 10a,b and 13a,b were prepared and employed in solid-phase oligonucleotide synthesis. For compounds 1a and 1b the lactime proton is protected to avoid unresolved degradation of its phosphoramidites 10a,b. UV-melting studies have been carried out to assess the thermal stability of oligonucleotides containing compounds 1a,b, and 3a,b. PMID- 16248048 TI - Propynyl groups in duplex, hairpin and triplex DNA: 7-deazapurines and 9 deazapurines. AB - Oligonucleotides containing 7-deazapurines or 9-deazapurines with propynyl groups at the 7- or 9-position were prepared. The stabilizing effect of the propynyl group was studied on DNA duplexes, hairpins and triplexes. PMID- 16248049 TI - Novel DNA nanoparticles and networks. AB - Joining the thrombin-binding aptamer 5-d(GGTTGGTGTGGTTGG) and the minihairpin 5 d(GCGAAGC) leads to new DNA nanoparticles, which are different from rod-like helical double-stranded DNA. Covalent interstrand cross-links in DNA duplexes generated by bifunctional alkadiyne chains were used to build-up the DNA networks. PMID- 16248051 TI - Parallel and antiparallel DNA: fluorescence quenching of ethidium bromide by 7 deazapurines. AB - The fluorescence quenching of ethidium bromide caused by 7-deaza-2' deoxyguanosine or 7-deaza2'-deoxyisoguanosine is observed in DNA with parallel or anti-parallel chain orientation. PMID- 16248050 TI - L-nucleosides containing modified nucleobases. AB - The synthesis of base modified L-nucleosides is described with pyrrolo[2,3 d]pyrimidines, pyrazolo[3,4-d]pyrimidines, benzimidazoles, and imidazo[1,2-a]-s triazines as nucleobases. The conformation of the nucleosides is studied and the antiviral activity is evaluated. PMID- 16248052 TI - 7-substituted 8-aza-7-deazapurines and 2,8-diaza-7-deaza-purines: synthesis of nucleosides and oligonucleotides. AB - 7-Substituted 8-aza- 7-deazaadenosines 1a-e were synthesized by Sonogashira cross coupling from the corresponding 7-iodo nucleoside in 36-79% yields. Starting from 7-bromo (or 7-iodo)-8-aza-7-deazaadenine, 2a,b were obtained by acid-catalyzed glycosylation followed by deprotection in 53 and 35% yields, repectively. Compounds 2b was applied to cross coupling reaction to give 2c-d in 34-95% yield. Compounds 2a and 4b were further transformed to the phosphoramidites 5 and 6b in 9 and 49% overall yields, which were incorporated into oligonucleotides. PMID- 16248054 TI - Synthesis and biological activity of 2-fluoro adenine and 6-methyl purine nucleoside analogs as prodrugs for suicide gene therapy of cancer. AB - A novel series of 6-methylpurine nucleoside derivatives with substitutions at 5 position have been synthesised These compounds bear a 5'-heterocycle such as triazole or a imidazole with a two carbon chain, and an ether, thio ether or amine. To extend the SAR study of 2-fluoroadenine and 6-methyl purine nucleosides, their corresponding alpha-linker nucleosides with L-xylose and L lyxose were also synthesized. All of these compounds have been evaluated for their substrate activity with E. coli PNP. PMID- 16248053 TI - Synthesis and in vitro anti-HCV activity of beta-D- and 1-2'-deoxy-2' fluororibonucleosides. AB - Based on the discovery of beta-D-2'-deoxy-2'-fluorocytidine as a potent anti hepatitis C virus (HCV) agent, a series of beta-D- and L-2'-deoxy-2' fluoroibonucleosides with modifications at 5 and/or 4 positions were synthesized and evaluated for their in vitro activity against HCV and bovine viral diarrhea virus (BVDV). The introduction of the 2'-fluoro group was achieved by either fluorination of 2,2'-anhydronucleosides with hydrogen fluoride-pyridine or potassium fluoride, or a fluorination of arabinonucleosides with DAST. Among the 27 analogues synthesized, only the 5-fluoro compounds, namely beta-D-2'-deoxy 2',5-difluorocytidine (5), had anti-HCV activity in the subgenomic HCV replicon cell line, and inhibitory activity against ribosomal RNA. As beta-D-N4 hydroxycytidine (NHC) had previously shown potent anti-HCV activity, the two functionalities of the N4-hydroxyl and the 2'-fluoro were combined into one molecule, yielding beta-D-2'-deoxy-2'-fluoro-N4-hydroxycytidine (12). However, this nucleoside showed neither anti-HCV activity nor toxicity. All the L-forms of the analogues were devoid of anti-HCV activity. None of the compounds showed anti BVDV activity, suggesting that the BVDV system cannot reliably predict anti-HCV activity in vitro. PMID- 16248055 TI - Aryl nucleoside H-phosphonates as a tool for investigation of stereospecificity during coupling. AB - It was found that in stereoselective condensations of ribonucleoside 3'-H phosphonates with alcohols, the major diastereomer of the produced H-phosphonate diesters is formed from the minor diastereomer of the intermediate phosphonic pivalic anhydride. PMID- 16248056 TI - Recent highlights on photolithic oligonucleotide array in situ synthesis. AB - Light directed synthesis of high-density oligonucleotide microarrays is currently performed using either ortho-nitro-benzyl-type [MeNPOC] (Pease, A.C.; Solas, D.; Sullivan, E.J.; Cronin, T.M.; Holmes, C.P.; Fodor, S.P.A. Proc. Natl. Acad Sci U.SA. 1994, 91, 6333.) or ortho-nitrophenylethyl-type [NPPOC] (Hasan, A.; Stengele, K.P.; Giegrich, H.; Cornwell, P.; Isham, K.R.; Sachleben, R.A.; Pfleiderer, W.; Foote, R.S. Tetrahedron 1997, 53, 424Z) protecting groups as the 5'-O-carbonate ester of the phosphoramidite building block. The synthesis cycle uses a combinatorial approach attaching one specific base per cycle, thus as many as 100 cycles need to be run to make an array of 25-mers. Time needed for deprotection/activation of the growing oligo chain determines overall manufacturing time and consequently also cost. In this report we demonstrate the development of photoprotected posphoramidite monomers for light directed array synthesis with increasing sensitivity to the UV light used. If combined with maskless array synthesis, this technology allows for synthesis of arrays with >780,000 different 25-mer oligonucleotides in about one hour and allows for high flexibility in array design and reiterative redesign. The arrays synthesized show high quality and reproducibility in our standard hybridization based assay. PMID- 16248057 TI - A solid supported reagent for internucleoside H-phosphonate linkage formation. AB - A fast and convenient procedure for synthesis of dinucleoside H-phosphonates is obtained through use of the novel polystyrene supported 5-carboxy-5-methyl-2-oxo 2-chloro-1,3,2-diaoxaphosphorinane reagent. Virtually quantitative H-phosphonate condensations are obtained leading to excellent isolatedyields and with only a simple filtration as the purification procedure. This provides for a convenient and high-yielding procedure that should be suited for solution-phase synthesis of oligonucleotides. PMID- 16248058 TI - Studies in oligonucleotide-based artificial nuclease systems. intramolecular copper (II) complex formation in an oligonucleotide bis-phenanthroline conjugate. AB - We have recently developed oligonucleotide based artificial nuclease (OBAN) systems based on 2'-O-methyloligoribonucleotides carrying a 2,9 dimethylphenanthroline x Zn(II) complex. These hybridize to an RNA molecule with bulge formation in the central region of the target and cleave the RNA target in a catalytic manner. When studying an 11-mer 2'-O-methyloligoribonucleotide carrying two 2,9-dimethylphenanthroline moieties, located 5 base pairs apart from each other, we found that this forms a cyclic structure in the presence of Cu2+ ions. This is due to intramolecular Cu(2,9-dimethylphenanthroline)2 complex formation, i.e., with the two ligands conjugated to the same oligonucleotide. PMID- 16248059 TI - A study of H-bonding of 3- and 5-substituted 6-aminouracils in duplex and triplex structures. AB - All possible dimers of the title modified bases with native nucleobases [10 dimers from 3-methylated 6-aminouracils (3sau) and 20 from 5-methyled 6 aminouracils (5sau), respectively have been calculated by ab initio method (Hartree-Fock method, 3 21G basis set). We have found two potential duplexes of 5sau and three possible duplexes of 3sau. Altogether seven dimers containing one or two bifurcating H-bonds have been found. Later on, five triplexes from ten possible calculated dimers have been found. In two of them the amino group of 6 aminouracil moiety takes part in H-bonding and there are H-bonds, too, between the first and third base of the triplexes causing an extra stabilization. PMID- 16248060 TI - Synthesis and biological activity of 2'-deoxy-4'-thio-pyrazolo[3,4-d]pyrimidine nucleosides. AB - The coupling of 4-aminopyrazolo [3, 4-d]pyrimidine with the appropriate thio sugar gave a 3:1 ratio of alpha,beta blocked 4-amino-1-(2-deoxy-4-thio-D erythropentofuranosyl)-1H pyrazolo[3,4-d]pyrimidine nucleosides. The mixture was deblocked, both the anomers were separated, and the beta-anomer was readily deaminated by adenosine deaminase. The nucleosides have been characterized, and their anomeric configurations have been determined by proton NMR. All three nucleosides were evaluated against a panel of human tumor cell lines for cytotoxicity in vitro. The details of a convenient and high yielding synthesis of these nucleosides are described. PMID- 16248061 TI - Novel strategy for single nucleotide polymorphism (SNP) genotyping by heteroduplex analysis: specific stabilization of TT mismatch base pair by mercury (II) cation and CC mismatch base pair by silver (I) cation. PMID- 16248062 TI - Oxidation of biotin during oligonucleotide synthesis. AB - A new "polystyrene biotin support" has been synthesized for the solid support synthesis of the 3'-biotinylated oligonucleotides. Several oligos were synthesized and were analyzed by the HPLC and Mass Spec. Oligo analysis revealed that the biotin gets oxidized to "biotin sulfoxide" during the synthesis. PMID- 16248064 TI - A new protecting group for the 5'-hydroxyl group having O-S single bond oxidatively cleavable under mild conditions. AB - We developed a new protecting group, ie., cis-[4-[[(4 methoxytrityl)sulfenyl]oxy]tetrahydrofuran-3-yl]oxycarbonyl (MTFOC), which could be removed under neutral conditions involving the oxidative removal of the MMTrS group followed by the self-cyclization of the resulting intermediate. The introduction of the protecting group into the 5-hydroxyl group of a thymidine derivative and its deprotection were studied. PMID- 16248063 TI - Synthesis and evaluation of anti-HSV activity of new 5-alkynyl-2'-deoxyuridines. AB - Eight 5-alkynyl-2'deoxyuridines containing different bulky substituents have been prepared and tested against HSV-1 in Vero cells. The compounds show positive antiviral activity. There is no obvious correlation between activity and substituent size. The nature of the linker between uracil and a substituent appears to be more important for antiviral properties: nucleosides containing arylethynyl groups show higher activity. PMID- 16248065 TI - Perylene attached to DNA through stiff or flexible linker: duplex stability and FRET. AB - Two fluorescent nucleosides, 5-(perylen-3-ylethynyl)-2'-deoxyuridine and 5 [(perylen-3-yl)methoxypropyn-1-yl]-2'-deoxyuridine, were incorporated into synthetic oligodeoxyribonucleotides and spectral properties of the conjugates and their duplexes were studied. PMID- 16248066 TI - 2-substituted 5'-N-methylcarboxamidoadenosine (MECA) derivatives as A3 adenosine receptor ligands. PMID- 16248067 TI - "Lock-in" modified cyclosal nucleotides--the second generation of cyclosal prodrugs. AB - A new generation of cycloSal-pronucleotides is presented. CycloSal-d4TMPs have been modified by introduction of an esterase-cleavable site in order to trap them inside cells. Hydrolysis studies in different media (PBS, CEM/0- and liver extracts) and anti-HIV evaluation of separated diastereomers revealed unexpected differences between the isomers. PMID- 16248068 TI - Incorporation of ribonucleoside 5'-(alpha-P-borano)triphosphates into a 20-mer RNA by T7 RNA polymerase. AB - The enzymatic synthesis of short boranophosphate RNA was studied by comparing the yield and pattern of abortive products of in vitro transcription at steady-state conditions with that of normal RNA. Boranophosphate short RNA can be readily synthesized by T7 RNA polymerase. PMID- 16248069 TI - Synthesis of 5-(1-propynyl)-2'-deoxyuridine 5'-(alpha-P-borano)triphosphate and kinetic characterization as a substrate for mmlv reverse transcriptase. AB - In order to introduce pyrimidine C5-propynyl modification into boranophosphate oligodeoxyribonucleotides (BP- ODNs), 5-(1-propynyl)-2'-deoxyuridine 5'-(alpha-P borano) triphosphate (d5PUTPalphaB) was synthesized. The two diastereomers were separated by reverse-phase HPLC. Kinetic studies showed that the Rp isomer was a slightly better substrate for MMLV reverse transcriptase than thymidine triphosphate or Rp-thymidine 5'-(alpha-P-borano)triphosphate. Using the Rp isomers of d5PUTPalphaB and the other three 5'-(alpha-P-borano) triphosphates, a DNA primer could be extended to the full length of the template. PMID- 16248070 TI - The effect of a single boranophosphate substitution with defined configuration on the thermal stability and conformation of a DNA duplex. AB - Substitution of one non-bridging oxygen in a natural phosphodiester internucleotide linkage with a borano (-BH3) group results in a chiral phosphorus center in boranophosphate. UV thermal melting profiles were recorded for DNA duplexes formed between a DNA 9-mer with either an Sp or a Rp boranophosphate linkage in the middle and the complementary DNA 9-mer, as well as for their unmodified parent duplex. The thermal stability of the DNA duplexes was in the order of normal > Sp borano > Rp borano. CD spectra of all three duplexes exhibited typical B-DNA profile, which closely resembled each other. PMID- 16248071 TI - Synthesis of N3,5'-cyclo-4-(beta-D-ribofuranosyl)-vic-triazolo[4,5-b]pyridin-5 one and its 3'-deoxysugar analogue as potential anti-hepatitis C virus agents. AB - We recently discovered a novel compound, identified as N3, 5-cyclo-4-(beta-D ribofuranosyl)-vic-triazolo[4,5-b]pyridinin-5-one, with anti-hepatitis C virus (HCV) activity in vitro. The structure was confirmed by chemical synthesis from 2 hydroxy-5-nitropyridine. It showed anti-HCV activity with EC50= 19.7 microM in replicon cells. Its 3'-deoxy sugar analogue was also synthesized, but was inactive against HCV in vitro. PMID- 16248072 TI - Synthesis and anti-hepatitis C virus activity of nucleoside derivatives of N3, 5' anhydro-4-(beta-D-ribofuranosyl)-8-aza-purin-2-ones. AB - A series of N3, 5-Anhydro-4-(beta-D-ribofuranosyl)-8-azapurin-2-ones were prepared in multistep reactions from uridine as potential anti-hepatitis C virus (HCV) agents. The synthetic details as well as biological evaluations are discussed. PMID- 16248073 TI - Synthesis and characterization of DNA duplexes containing an N3T-ethyl-N3T interstrand crosslink in opposite orientations. AB - DNA duplexes containing an ethyl interstrand crosslink that bridges the N3 atoms of thymidines on the opposite strands have been synthesized using an approach that combines conventional solid phase oligonucleotide synthesis and the selective removal of protecting groups of a crosslinked thymidine dimer. This approach allows for the assembly of a crosslinked duplex directly on the solid support. Duplexes that contain a N3T-ethyl-N3T interstrand crosslink in a staggered orientation at either a -TA- or -AT-step in a duplex have been prepared. When placed in an -AT- step of a duplex the effect was stabilizing relative to the non-crosslinked control duplex (deltaTm= +24 degrees C) and this crosslinked duplex was found to efficiently form multimers in the presence of T4 ligase. In the case of the -TA- crosslinked duplex the stabilizing effect was less pronounced (deltaT.= +6 degrees C) and likewise did not undergo self ligation under identical conditions. Molecular modeling studies suggested that the -AT- containing lesion had little deviation in structure relative to the non crosslinked duplex DNA control, whereas the -TA- crosslinked duplex exhibited significant buckling of the base pairs flanking the lesion. PMID- 16248074 TI - Antigene property of PNA conjugated to the nuclear localization signal peptide. AB - Peptide nucleic acid (PNA) is a DNA mimic with antigene properties. To enhance its capacity to enter in the cell and internalize in the nucleus, PNA has been conjugated to the nuclear localization signal (NLS) peptide, PKKKRKV PNA-NLS conjugates form stable hybrids with complementary DNA strands and poorly tolerate mismatched base pairing. Employed against cancer-associated genes, PNA-NLS exhibited a potent and specific antigene activity, suggesting exciting therapeutic approaches to cancer. PMID- 16248075 TI - Synthesis of 3'-deoxy-3'-C-hydroxymethyl analogues of tiazofurin and ribavirin. AB - On the basis of potent biological activity of 3'-branched-3'-deoxynucleoside analogues, novel ribavirin and tiazofurin derivatives with 3'-C-hydroxymethyl substituent were synthesized, starting from D-xylose. PMID- 16248076 TI - Synthesis of homo-N-nucleoside with 1,2,4-triazole-3-carboxamide. AB - On the basis of potent biological activities of ribavirin and homo-N-nucleosides, a novel homo-N-1,2,4-triazole-3-carboxamide derivative 1 was synthesized starting from 2,3,5-tri-O-benzoylribofuranosyl-1-acetate as a potential antiviral agent. PMID- 16248077 TI - Affinity of (alpha-P-borano)-NTP analogs to rabbit muscle pyruvate kinase. AB - The binding affinity of (alpha-P-borano) and other NTP analogs to rabbit muscle pyruvate kinase (PK) was investigated using a fluorescence quenching approach to obtain structure-activity relationships for substrate specificity of nucleotide analogs. PMID- 16248078 TI - A novel amino-on CPG-support for the synthesis of 3'-aminoalkylated oligonucleotides. AB - The synthesis of a novel amino-ON CPG support and its application in the synthesis of 3'-aminoalkylated oligonucleotides is reported. The release of oligonucleotides with free 3'-amino groups is accomplished by treatment with concentrated ammonia for 2 h at 55 degrees C. PMID- 16248079 TI - Enzymatic resolution and base pairing properties of D- and L-cyclohexenyl nucleic acids (CeNA). AB - An enzymatic transesterification reaction afforded large scale resolution of the cyclohexenol precursor needed for preparation of both series of CeNA building blocks. CeNA oligos of "D-like" chirality display a strong and selective interaction with RNA, while preserving RNase H activity, and therefore have potential as antisense constructs. CeNAs of opposite chirality form a self pairing system on their own. PMID- 16248080 TI - Photolabeling probes of ribavirin and EICAR. AB - Ribavirin, the only small molecule available so far for treating hepatitis C virus infection, was recently used in an emergency context to treat patients with severe acute respiratory syndrome in the early stages of the disease. EICAR, one of the most potent congeners of ribavirin, has 10 to 100 times greater antiviral potency than ribavirin. The mechanisms underlying the antiviral effects of ribavirin and EICAR have not yet been definitely elucidated, but they seem to be similar. In order to study the mechanisms responsible for their antiviral effects using a photolabeling approach, we have developed photolabeling probes of ribavirin and EICAR, in which an azido group was introduced into the pseudobases of triazole and imidazole, respectively. The ribavirin photoprobes were obtained by directly coupling the azidotriazole to the protected ribose sugar, while the EICAR probe was prepared by diazotizing AICAR and subsequently substituting with NaN3. All these probes showed a fast, clear-cut photochemical reaction, which suggests that they are promising tools for use in photolabeling studies. PMID- 16248081 TI - Silyl protecting groups for oligonucleotide synthesis removed by a ZnBr2 treatment. AB - An oligonucleotide protected with N-(trimethylsilyloxycarbonyl) (Teoc) and P (trimethylsilylethanol) (Tse) groups was synthesized and deprotected by a single ZnBr2 treatment. Finally it was released from the solid support by cleavage of a disulfide linkage with TCEP. The olgonucleotide was obtained without any basic treatment. PMID- 16248082 TI - Synthesis of conformationally restricted nucleic acid fragments using ring closing alkene and enyne metathesis reactions. AB - In the aim of constructing conformationally restricted nucleic acid fragments for the recognition of secondary RNA structures, we have synthesized different mono- and dinucleotides containing extra rings. These rings were prepared by ring closing alkene or enyne metathesis reactions from nucleotide substrates in which double or triple bonds have been introduced. PMID- 16248083 TI - Promotion of pyrimidine motif triplex formation by morpholino modification of triplex-forming oligonucleotide: kinetic and thermodynamic studies. PMID- 16248084 TI - RNA recognition by fluor-aromatic substituted. AB - RNA exhibits a higher structural diversity than DNA and is an important molecule in the biology of life. It shows a number of secondary structures such as duplexes, hairpin loops, bulges, internal loops, etc. However, in natural RNA, bases are limited to the four predominant structures U, C, A, and G and so the number of compounds that can be used for investigation of parameters of base stacking, base pairing, and hydrogen bond is limited. We synthesized different fluoromodifications of RNA building blocks: 1'-deoxy-1'-phenyl-beta-D ribofuranose (B), 1'-deoxy-1'-(4-fluorophenyl)-beta-D-ribofuranose (4 FB), 1' deoxy-1'-(2,4-difluorophenyl)-beta-D-ribofuranose (2,4 DFB), 1'-deoxy- 1'-(2,4,5 trifluorophenyl)-beta-D-ribofuranose (2,4,5 TFB), 1'-deoxy- 1'-(2,4, 6 trifluorophenyl)-beta-D-ribofuranose, 1'-deoxy- 1'-(pentafluorophenyl)-beta-D ribofuranose (PFB), 1'-deoxy-1'-(benzimidazol-1-yl)-beta-D-ribofuranose (BI), 1' deoxy-1'-(4-fluoro-1H-benzimidazol-1-yl)-1-beta-ribofuranose (4 FBI), 1'-deoxy- 1'-(6-fluoro- 1H-benzimidazol-1-yl)-beta-D-ribofuranose (6FBI), 1'-deoxy- 1'-(4, 6-difluoro- 1H-benzimidazol- 1-yl)-beta-D-ribofuranose (4,6 DFBI), 1'-deoxy- 1' (4-trifluoromnethyl- H-benzimidazol-1-yl)-beta-D-ribofuranose (4 TFM), 1'-deoxy 1'-(5-trifluoromnethyl-1H-benzimidazol-1-yl)-beta-D-ribofuranose (5 TFM), and 1' deoxy-1'-(6-trifluoromethyl-1H-benzimidazol-1-yl)-beta-D-ribofuranose (6 TFM). These amidites were incorporated and tested in a defined A, U-rich RNA sequence (12-mer, 5-CUU UUCXUU CUU-3' paired with 3'-GAA AAG YAA GAA-5'). Only one position was modified, marked as X and Y, respectively. UV melting profiles of those oligonucleotides were measured. PMID- 16248085 TI - Prosit, an online service to calculate pseudorotational parameters of nucleosides and nucleotides. AB - The online service PROSIT (Pseudo-Rotational Online Service and Interactive Tool) is a free service available at http://cactus.nci.nci.gov/prosit/ that performs pseudorotational analysis of nucleosides(tides). PROSIT reads the 3D coordinates of nucleosides and returns the pseudorotational phase angle P, puckering amplitude vmax and other related information. As examples, the sugar conformations in a parallel-stranded guanine tetraplex and a four-way Holliday junction are presented here. PMID- 16248086 TI - A cautionary note on the use of the 31P NMR spectroscopy in stereochemical correlation analysis. AB - Stereoselectivity in condensation of protected ribonucleoside 3'-H-phosphonates with hydroxylic components was investigated using 31P NMR spectroscopy. The correlation between absolute configuration at the phosphorus center and the chemical shifts of the produced H-phosphonate diesters and the corresponding phosphorothioates, was studied. PMID- 16248087 TI - Significant improvement of quality for long oligonucleotides by using controlled pore glass with large pores. AB - A key factor influencing the quality of long oligonucleotides is the choice of controlled pore glass (CPG) which is used as a solid support during oligonucleotide synthesis. We studied the influence of CPG pore size on the quality of 75-mer oligonucleotides. Using electrophoresis and HPLC, we demonstrated failure modes that can occur at certain oligo lengths with 1000A pore size, and compared yield and purity of 75-mer oligos using 1000A and larger pore size CPG. We showed that oligonucleotides with much better quality are obtained using CPG with pore sizes of 1400A and larger. We also identified the key characteristics for CPG selection that lead to the best CPG performance. PMID- 16248088 TI - Functionalization of pyrimidine and purine nucleosides at C4 and C6: C nucleophilic substitution of their C4- and C6-(1,2,4-triazol-1-yl) derivatives. AB - A study of C-nucleophilic substitution at the C4-position on pyrimidine and C6 position on 2'-deoxyguanosine to produce novel nucleosides is presented with the spectroscopic properties of their respective substitution products. C4-(1,2,4 triazol-1-yl) pyrimidine nucleosides 1 were treated with nitroalkanes, malononitrile, acetylacetone, ethyl nitroacetate, acetoacetate and cyanoacetate at 100 degrees C in dioxane in the presence of DBU resulting in the production of novel nucleosides 2-11. To explore the application of this methodology to purine chemistry, this approach was used to produce novel analogs from 2' deoxyguanosine. We found that the triazolo derivative 12 undergoes C-nucleophilic substitution with nitromethane, malononitrile, acetylacetone, ethyl nitroacetate and cyanoacetate in the presence of potassium carbonate (K2CO3) in DMF at 100 degrees C to give novel nucleosides 13-17. PMID- 16248089 TI - Synthesis of 5-ethynyl-2'-deoxyuridine-5'-boranomono phosphate as a potential thymidylate synthase inhibitor. AB - The 5-ethynyl-2'-deoxyuridine nucleoside and the 5'-boranomonophosphate nucleotide were synthesized as analogs of 5-fluoro-2'-deoxyuridine monophosphate (5-FdUMP), a widely used mechanism-based inhibitor of thymidylate synthase. Synthesis was carried out from protected 5-iodo-2'-deoxyuridine and trimethylsilylacetylene by Sonogashira palladium-catalyzed cross coupling reaction followed by selective phosphorylation and finally boronation. PMID- 16248090 TI - Synthesis study of 3'-alpha-fluoro-2',3'-dideoxyguanosine. AB - A synthetic method was established for 3'-alpha-fluoro-2','3-dideoxyguanosine 1 from guanosine 2 in 27% overall yield and 6 steps. A byproduct 6a of fluorination was identified by NMR studies, its presence strongly supporting our supposition that the fluorination itself proceeded via a bromonium cation. PMID- 16248091 TI - Presence of 2',5'-linkages in a homopyrimidine DNA oligonucleotide promotes stable triplex formation under physiological conditions. AB - We prepared 15-mer homopyrimidine oligonucleotides containing three or four 2',5' linked DNA units, and their ability as a triplex-forming oligonucleotide (TFO) was analyzed in detail UV melting experiments showed that replacement of a 3',5' linkage by a 2',5'-linkage at every third or fourth residue in TFO significantly promoted stable triplex formation under physiological conditions. PMID- 16248092 TI - A convenient method for the synthesis of oligonucleotide-cationic peptide conjugates. AB - A method was developed for the synthesis of oligonucleotide-cationic peptide conjugates in solution phase by disulfide bond formation. Precipitation was avoided by the easily removable triethylammonium trifluoroacetate (TEATFAc) salt which served at the same time as a buffer of the reaction mixture. The fast and high yielding disulfide bond formation was due to the Npys thio protecting and activating group of Cys. A solution of the free 5-thiol modified oligonucleotide obtained from Poly-Pak purification was used for conjugation. PMID- 16248093 TI - A new approach to oligonucleotide N3'-->P5' phosphoramidate building blocks. AB - A new synthetic approach to 5-phosphoramidites of 3'-aminonucleosides was developed. The methodology relies upon the use of 3'-amino-2',3'-dideoxy nucleosides as the key starting materials. The final phosphoramidite products were obtained with high yields via 2-3-step efficient chemical transformations using selective introduction of orthogonal protective groups to the 3' aminonucleoside sugar and base moieties. PMID- 16248094 TI - Oligonucleotides as coding molecules in an anti-counterfeiting system. AB - Due to the growing numbers of counterfeited products on the world market, there is a huge demand for new and forgery-proof marking systems. We developed a unique system using "molecular beacons" with well adapted thermodynamic parameters. This marking system consists of the three components: DNA tag (a label or directly printing), detection pen (contains the "molecular beacon " solution), and DNA scanner (reads the specific signal triggered by the detection pen even at daylight). The vast coding capacity of DNA combined with the highly specific signal offers a degree of security that is unmatched by conventional identification technologies. PMID- 16248095 TI - Studies on the attachment of DNA to silica-coated nanoparticles through a Diels Alder reaction. AB - A new method has been investigated for the functionalization of gold nanoparticles with DNA. Silica-coated nanoparticles functionalized with a maleimide have been prepared. These particles are designed to react with modified DNA containing a diene functionality at one end of the molecule. The result would be the formation of a more stable attachment of the DNA to the particle through a Diels-Alder reaction. This covalent attachment would not be susceptible to ligand exchanges, which are known to occur in the conventional DNA functionalization of gold nanoparticles. PMID- 16248096 TI - Chemical synthesis of oligonucleotides containing (M+4) guanine. AB - HPLC MS/MS has shown great potential in the measurement of DNA oxidative damage. Its accuracy depends on the use of multiply isotopically labelled internal standards. In this report, multiply isotopically labelled (M + 4) guanine internal standards were prepared in the form of base, nucleoside, as well as DNA oligomer. To our knowledge, this is the first chemical synthesis of oligomers containing (M + 4) guanine, and we believe that they can be used to develop a procedure that can make further improvement to the existing analytical procedures. PMID- 16248097 TI - Tumor-targeting peptide-PNA-peptide chimeras for imaging overexpressed oncogene mRNAs. AB - We have optimized a method involving continuous solid phase synthesis of chelator peptide-PNA-peptide probes in order to noninvasively image oncogene mRNAs overexpressed in tumors. The PNA (peptide nucleic acid) probes carry cyclized peptide ligand analogs specific for receptors overexpressed on malignant breast or colorectal cancer cells, and chelators to bind radioactive metal ions, or a fluorophore. In vivo scintigraphic imaging of MCF7 xenografts in immunocompromised mice indicated that CCND1 and MYC [99sTc] chelator-PNA-D (CSKC) probes concentrated in MCF7 cells up to 7 times more than the corresponding mismatch controls. PMID- 16248098 TI - Natural phosphate doped with KI in HMDS: a mild and efficient reagent for alkylation and glycosylation of nucleobases. AB - Several D-ribonucleosides are prepared from 1-O-acetyl-2,3,5-tri-O-benzoyl-beta-D ribofuranoside and trimethylsilylated nucleobases under mild conditions by using natural phosphate doped with KI as catalyst. PMID- 16248099 TI - Synthesis and properties of a novel bridged nucleic acid analogue, 5'-amino-3',5' BNA. AB - An oligonucleotide P3'-->N5' phosphoramidate (5'-amino-DNA) attracts much attention because of its potential for application to DNA sequencing; however, its ability to hybridize with complementary strands is low. To overcome this drawback of the 5-amino-DNA, we have designed and successfully synthesized a novel nucleic acid analogue having a P3'-->N5' phosphoramidate linkage and a constrained sugar moiety, 5'-amino-3'-C,5'-N-methylene bridged nucleic acid (5' amino-3',5'-BNA). The binding affinity of the 5'-amino-3',5'-BNA towards complementary DNA and RNA strands was investigated by UV melting experiments. The melting temperature (Tm) of the duplex comprising the 5'-amino-3',5'-BNA and its complementary strand was much higher than that of the duplex containing the corresponding 5-amino-DNA. PMID- 16248100 TI - Synthesis of a 2'-amino-alpha-1-LNA-T phosphoramidite. AB - A convergent route to 2'-amino-alpha-L-LNA-T phosphoramidite building block 16 has been developed. Key steps include 1) introduction of a C2-azido group prior to nucleobase-coupling, 2) tandem Staudinger and intramolecular nucleophilic substitution reaction, and 3) separation of alpha-L- and beta-L-configured intermediates. PMID- 16248101 TI - Modified binary hammerhead ribozymes with high catalytic activity. AB - A series of binary hammerhead ribozymes was designed and assessed in terms of cleavage activity and nuclease resistance. Enhanced nuclease resistance of binary ribozymes was achieved by incorporation of Z-modified nucleotides at the selective positions along with addition of 3'-3-linked thymidine cap. These modified binary ribozymes efficiently cleave 190-nucleotides long MDR1 mRNA fragment and display catalytic activity much higher then respective full-length analogs. PMID- 16248102 TI - 4,5-bis(ethoxycarbonyl)-[1,3]dioxolan-2-yl as a new orthoester-type protecting group for the 2'-hydroxyl function in the chemical synthesis of RNA. AB - We wish to report 4,5-bis(ethoxycarbonyl)-[1,3]dioxolan-2-yl as a new protecting for the 2'-hydroxyl function. Our cyclic orthoester-type group is compatible with the DMTr strategy for oligonucleotide synthesis. This group was introduced to the 2'-hydroxyl group of appropriately protected nucleoside derivatives in good yields under mild acidic conditions. Post-synthetic conversion of the moiety of this protecting group with an amine resulted in formation of a new amide moiety that is more stable to acid deprotection in aqueous solution, but it can still be easily removed by treatment with acids in organic solvents. In this article, we also describe the stability of not only the original and modified protecting groups but also internucleotidic phosphate linkages of protected RNA intermediates under deprotection conditions. PMID- 16248103 TI - Stereoselective synthesis of novel thioiso dideoxy nucleosides with exocyclic methylene as potential antiviral agents. AB - Novel thioiso pyrimidine and purine nucleosides substituted with exocyclic methylene have been synthesized, starting from D-xylose. Cyclization of the dimesylate to the 4-thiosugar 6a proceeded in pure SN2 reaction in the presence of allylic functional group. PMID- 16248104 TI - Synthesis of 3'-ureidoadenosine analogues and their binding affinity to the A3 adenosine receptor. AB - Novel 3'-ureidoadenosine analogues were synthesized from 1,2:5, 6-di-O isopropylidene-D-glucose in order to lead to stronger hydrogen bonding at the A3 adenosine receptor than the corresponding 3'-aminoadenosine derivatives. However, all synthesized 3'-ureidoadenosine analogues have lost their binding affinities to the all subtypes of adenosine receptors, indicating that bulky 3'-urea moiety led to conformational distortion. PMID- 16248105 TI - Synthesis and properties of PNA oligomers containing orotic acid derivatives. AB - We have investigated the incorporation of C6-derivatives of uracil into polypyrimidine peptide nucleic acid oligomers (PNA). Starting with orotic acid (uracil-6-carboxylic acid) we have prepared a PNA monomer containing the methyl orotate nucleobase which is compatible with Fmoc-based synthesis. Treatment of the resin-bound oligomers with hydroxide or amines cleanly converted the ester to an orotic acid or orotamide-containing PNA. Alternatively, the methyl orotate containing PNA was liberated from the resin by standard acidolysis. PNA bearing a modified nucleobase was found to hybridize to both poly(rA) and poly(dA). Complexes with poly(rA) were more stable than those with poly(dA) but both were destabilized relative to an unmodified PNA. Modification of a terminal residue was tolerated better than modification of an internal position. The type of charge provided by the modification affected the complex stability. In the worst case, an internal modification was nearly as detrimental as a base mismatch. PMID- 16248106 TI - Efficient chemo-enzymatic syntheses of pharmaceutically useful unnatural 2' deoxynucleosides. AB - Our chemo-enzymatic method was successfully applied to the synthesis of 2-chloro 2'-deoxyadenosine (CdA, cladribine) in two ways: 1) direct conversion of chemically synthesized 2-deoxy-alpha-D-ribose 1-phosphate (dRP) to CdA; 2) a two step route via 9-(2-deoxy-beta-D-ribos-1-yl)-2, 6-dichloropurine (Cl2Pu-dR, 5). PMID- 16248109 TI - A recommendation to biomedical engineering students. PMID- 16248107 TI - Novel way of capping mRNA trimer and studies of its interaction with human nuclear cap-binding complex. AB - Binding of mRNA 5' cap by the nuclear cap-binding complex (CBC) is crucial for a wide variety of mRNA metabolic events. The interaction involving the CBP20 subunit of CBC is mediated by numerous hydrogen bonds and by stacking of the tyrosine sidechains with two first bases of the capped mRNA. To examine a possible role of a longer mRNA chain in the CBC-cap recognition, we have synthesized an mRNA tetramer using a novel way of capping an RNA trimer and determined its affinity for CBC by fluorescence titration. PMID- 16248108 TI - Large-scale synthesis of 5'-O-pixyl protected 2'-deoxynucleosides useful for oligonucleotide synthesis. AB - Synthesis of 5'-O-pixylated 2'-deoxynucleosides 4 has been accomplished and the products are commercially available. PMID- 16248110 TI - Society rewards and competition awards. PMID- 16248111 TI - Counting to ten and reaching out. PMID- 16248112 TI - Old idea, new technology. PMID- 16248113 TI - A biomimetic retinal stimulating array. PMID- 16248114 TI - Silicon microsystems for neuroscience and neural prostheses. PMID- 16248115 TI - Restoring lost cognitive function. PMID- 16248116 TI - The functional reanimation of paralyzed limbs. PMID- 16248117 TI - Microelectronic packaging for retinal prostheses. PMID- 16248118 TI - Implantable biomedical microsystems for neural prostheses. PMID- 16248120 TI - Thermal effects of bioimplants. PMID- 16248119 TI - Implantable biomimetic microelectronic systems design. PMID- 16248121 TI - Turning students into science stars. PMID- 16248122 TI - Primary germ cell tumors of the mediastinum: 10 years of experience in a tertiary teaching hospital. AB - Germ cell tumors occur mostly in the gonad. Extragonadal germ cell tumors are rare, and most occur in the retroperitoneum and mediastinum. Primary mediastinal germ cell tumors are often found in the anterior portion of the mediastinum and include teratomas and non-teratomatous tumors. Non-teratomatous tumors include seminomas and malignant non-seminomatous germ cell tumors (MNSGCTs). MNSGCTs include yolk sac tumors, choriocarcinomas, embryonal carcinomas, and mixed type germ cell tumors. Teratomas are the most common germ cell tumors of the mediastinum, and seminomas are the most common non-teratomatous germ cell tumors of the mediastinum. Cases of primary mediastinal MNSGCT reported in the literature are rare. In this report, we review all primary mediastinal germ cell tumors from a 10-year period at the Chung-Ho Memorial Hospital of Kaohsiung Medical University. A total of 14 cases were reviewed, including 11 patients with mature teratomas, two with yolk sac tumors, and one with seminoma. We discuss the differences in clinical presentation, histopathologic characteristics, treatment, and prognosis. PMID- 16248123 TI - Initial intravenous gamma-globulin treatment failure in Iranian children with Kawasaki disease. AB - The purpose of this study was to determine the initial rates of intravenous gamma globulin treatment (IVIG) failure in Kawasaki disease (KD) and their predisposing factors. This study was a retrospective analysis of the initial response to IVIG (2 g/kg), assessed from the medical reports of all patients admitted to Namazee Hospital pediatric ward, from March 1998 to March 2002, and who fulfilled the criteria for KD. Data were available for 64 patients, 58 of whom (90.6%) became afebrile 48 hours after completion of the initial dose of IVIG (Group I) and six (9.4%) who remained febrile (Group II). Two patients had a prompt response to a second dose of IVIG. In Group I, five patients (8.6%) developed coronary artery disease, seen on echocardiography. In Group II, two patients (33.3%) developed coronary artery disease. No significant difference was found in the prevalence of coronary artery disease between the two groups (p = 0.12), or in age or gender. The rate of initial treatment failure was 9.4% in this cohort of patients, which is comparable with previous reports. No predictive factors such as coronary artery disease, age or gender were found for initial treatment failure in KD. PMID- 16248124 TI - Structural abnormality on brain magnetic resonance imaging in late-onset major depressive disorder. AB - The purpose of this study was to examine the structural abnormalities of patients with late-onset major depressive disorder using brain magnetic resonance imaging (MRI) and to assess clinical correlates of these structural abnormalities. Thirty seven elderly patients with DSM-IV major depressive disorder that first occurred after the age of 50 years, and 18 control subjects without depression were recruited. All participants underwent comprehensive psychiatric assessment and cerebral MRI. Brain ventricular and sulcal sizes and white matter hyperintensities were assessed visually. Relative to control subjects, patients with late-life major depressive disorder showed more severe brain atrophy (p = 0.043) and white matter hyperintensities (p = 0.024), especially in the periventricular area (p = 0.012). Over 60% of the patient group had significant brain MRI hyperintensities. White matter hyperintensity was correlated with later onset of depressive illness (r = 0.49, p = 0.002) among patients. Brain atrophy and white matter hyperintensities are prevalent in patients with late-onset major depressive disorders. These two abnormalities may represent different pathophysiologic processes of depressive disorders. White matter hyperintensities may be predisposing factors for late-onset major depressive disorder. PMID- 16248125 TI - Evaluation of intrarenal blood flow by Doppler ultrasonography immediately after extracorporeal shock wave lithotripsy on hydronephrotic kidney. AB - Extracorporeal shock wave lithotripsy (ESWL) is an effective and relatively noninvasive mode of treatment for urinary calculi. The aim of this study was to test whether therapeutic ESWL induces changes in renal parenchymatous blood flow and to evaluate shock wave side effects on the renal parenchyma. A total of 45 patients who underwent ESWL for ureteropelvic stone between January 2002 and July 2003 were included in this prospective study. Color Doppler sonography before and 30 minutes after ESWL showed no significant morphologic change. Resistive index (RI) was used to estimate renovascular resistance. The RI significantly increased in obstructed hydronephrotic kidneys. However, no significant change was observed in both treated and untreated kidneys before and after treatment. Hydronephrotic kidneys do not have a higher risk of post-ESWL renovascular resistance interference. The measurement of changes in RI with Doppler ultrasonography may provide useful information for clinical diagnosis of renal tubulointerstitial and vascular damage. PMID- 16248126 TI - Hydatid cyst of the liver: a case report and literature review. AB - Taiwan is nearly free from hydatid disease. We report a case of hydatid cyst of the liver in a 37-year-old man who originally lived in India and had migrated to Taiwan 2 years earlier. He presented with right upper quadrant pain and intermittent low-grade fever. Both sonography and computed tomography (CT) demonstrated a cystic lesion with vesicles at its periphery in segments 6 and 7 of the liver. A hydatid cyst was diagnosed. The patient underwent radical excision of the cyst with total removal without opening the wall. He also received pre- and postoperative oral mebendazole. Pathology showed a hydatid cyst consisting of three layers: the inner single nucleated geminal layer, the middle acellular laminated layer, and the outer pericyst originating from inflammatory and hepatic cells. This case highlights that accurate preoperative diagnosis of hydatid disease can be made from personal history, typical sonography and CT study in non-endemic areas. PMID- 16248127 TI - Sparing surgery for an epiderom cyst of the testis: a case report and literature review. AB - Epidermoid cyst of the testis is relatively rare and accounts for less than 1% of all testicular neoplasms. This condition usually occurs in young men and it is mostly detected by self-examination or routine physical examination. The case of a 27-year-old male with an epidermoid cyst of the testis is discussed. The diagnosis was based on ultrasonography and management was with testicular-sparing surgery. At 2-year follow-up, the patient had had no relapse or metastasis. In epidermoid cysts of the testis, high-frequency ultrasonography is a reliable diagnostic imaging method. Organ-preserving surgery is recommended in the management of these lesions. PMID- 16248128 TI - Huge spinal extradural meningeal cyst in the thoracolumbar spine: a case report of a rare cause of low backpain. AB - Symptomatic intraspinal extradural meningeal cysts are rare. We present the case of a 17-year-old female with low back pain, progressive numbness, and radiation pain to the flank and lower limbs. Magnetic resonance imaging and computed tomographic myelography revealed a huge intraspinal extradural meningeal cyst extending from T12 to L3 with cord and dural sac compression. The patient underwent surgery to open the cyst and close the connecting dural defect. Pathologic examination of the cyst disclosed non-specific fibrous connective tissue without an inner arachnoid single-cell lining. She achieved complete recovery after the operation. There was no recurrence of the cyst at a 2-year follow-up. PMID- 16248129 TI - Circumscribed choroidal hemangioma in a Chinese patient treated using transpupillary thermotherapy: a case report. AB - We describe the first case of a Chinese patient with circumscribed choroidal hemangioma (CCH) effectively managed using transpupillary thermotherapy (TTT). A 57-year-old man had an elevated orange-red subretinal mass with a base of 5 x 5 mm superior to the optic disc, along with serous macular detachment in the left eye. Ultrasonography depicted a mass 3 mm thick with acoustic solidity and high internal reflectivity. Fluorescein angiography showed an area of hyperfluorescence in the prearterial phase and tumor staining in the late phase, consistent with a diagnosis of CCH. Therefore, TTT (spot size, 3 mm; exposure, 1 min) was performed with a diode laser, delivered through a slit-lamp biomicroscope. The laser power was first set at 300 mW and then increased in 50 mW increments, until test shots in the mid-peripheral retina outside the lesion produced a slightly grayish appearance in the irradiated area. The tumor was covered with four overlapping laser spots and subjected to an average beam power of 400 mW. Best-corrected visual acuity increased from 20/50 before treatment, to 20/20 3 months following one session of TTT. The lesion became atrophic and the subretinal fluid disappeared. Our experience shows that TTT can be an even more effective treatment strategy for CCH in Asians than in Caucasians. In Chinese patients, the power level of the diode laser should be lower than that used in Caucasians. PMID- 16248130 TI - Meeting the challenge of reducing waiting times for surgery. PMID- 16248131 TI - Incidence of portal vein thrombosis after laparoscopic splenectomy. AB - BACKGROUND: Laparoscopic splenectomy has become an important therapeutic option in the management of diverse hematologic disorders. However, the incidence of important complications, such as portal vein thrombosis, remains poorly understood. We set out to study the incidence of this complication over a 14 month period. METHODS: All adult patients who underwent laparoscopic splenectomy between July 2001 and April 2002 at McMaster University Medical Centre in Hamilton, Ont., were approached postoperatively and offered duplex ultrasonography of the portal vein to look for thrombosis. RESULTS: During the study period 17 patients underwent laparoscopic splenectomy. Two patients declined to participate, and in 1 patient the ultrasonography was technically inadequate. Of the 14 remaining patients, 2 (14%) were found to have portal vein thrombosis. In 1 other patient, the investigation was suggestive, but not conclusive, of a clot in the portal system. CONCLUSIONS: Portal vein thrombosis in this series was a relatively common complication of laparoscopic splenectomy. Further research is needed to confirm our findings. PMID- 16248132 TI - Waiting times and patient perspectives for total hip and knee arthroplasty in rural and urban Ontario. AB - BACKGROUND: The demand for total hip and total knee arthroplasties is increasing as are the waiting times for these procedures. Because of the differences between rural and urban areas in terms of the provision of arthroplasty services and between the 2 patient groups, patient perspectives of waiting times may also be different. METHODS: To compare waiting times for initial orthopedic consultation and total hip and knee arthroplasties in rural Ontario (Stratford) and in urban Ontario (London), and to compare patient perspectives of these waiting times, we mailed a survey to all 260 patients who underwent total hip or total knee arthroplasty between June 1, 2000, and June 1, 2001. The survey asked for the length of wait for consultation and for surgery, acceptability of waiting time for surgery, the effect of waiting on health and what an acceptable waiting time would be. Of the 260 surveys mailed 202 (78%) were returned. We reviewed the charts of the respondents to determine the actual waiting times. RESULTS: The actual waiting times (mean [and standard deviation]) for initial consultation were significantly (p < 0.001) shorter in the rural (RUR) group (1.10 [0.53] mo) than the urban (URB) group (3.40 [1.34] mo). There was no significant difference in waiting times for surgery between RUR (8.45 [3.32] mo) and URB (9.32 [3.61] mo) groups. Surgical waiting times for both groups showed that 56% of all the patients had to wait longer than 9 months from the date surgery was recommended. Perceived waiting times for consultation were found to be 56.9% longer (p < 0.001) than the actual waiting times, but there was no significant difference between perceived and actual waiting times for surgery (p = 0.40). Fifty percent of the patients stated they were unhappy with the wait for surgery or found the wait unacceptable (56% of URB and 44% of RUR patients). There was no significant difference between RUR and URB in patients' acceptance of their wait for surgery (p = 0.09), but URB patients believed their wait for surgery made a greater contribution to health deterioration (p = 0.003). Thirty-eight percent of RUR and 54% of URB patients believed their surgical wait contributed to "a lot" or a "moderate" amount of deterioration in their health. CONCLUSIONS: Waiting times for hip or knee replacement surgery have increased to the point at which over 50% of surgical patients in 2000-2001 in RUR and URB orthopedic practices had waited longer than 9 months for surgery. In comparing these practices, there was a significantly longer wait in urban than rural practices for the initial consultation but no significant difference in waiting times for surgery. PMID- 16248133 TI - Comorbidity and age are both independent predictors of length of hospitalization in trauma patients. AB - BACKGROUND: Length of hospitalization is a good indicator of resource utilization. Older patients are increasingly suffering trauma, and comorbid medical conditions are also increasing. Our objective was to determine the separate and combined effect of these 2 factors on length of hospital stay for trauma patients in a tertiary trauma centre. METHODS: All 994 consecutive trauma patients surviving to hospital discharge between Apr. 1, 2000, and Mar. 31, 2001, were identified. Patient characteristics, injury severity and length of hospitalization were obtained from the hospital trauma registry. Each medical record was then reviewed for completeness of information and assessment of comorbid conditions. A multivariate linear regression model was developed to predict logarithmic length of stay from age and presence of a cormorbid condition while adjusting for the Injury Severity Score (ISS). RESULTS: The mean age of the patients was 49.7 (range from 14-100) years and median ISS was 9 (range from 1 50). At least 1 comorbid condition was present in 321 (32%) patients. Mean length of hospital stay was 15.3 days. The proportion of patients with a comorbid condition increased steadily with age, from 8.7% before the age of 55 years to 92% at 85 or more years of age (p < 0.001). According to the multivariate model, the presence of comorbidity, age and ISS were all independent predictors of hospital stay (p < 0.001). When applied to patients with the mean ISS value of 9, the model showed an increase in length of hospitalization for patients with a comorbid condition over those without; (3.6 v. 13.1 d for patients < 55 and > or = 85 yr respectively). Length of hospital stay increased particularly with neurologic and pulmonary problems. CONCLUSIONS: Comorbidity and age were both independently significant predictors of length of hospitalization over and beyond that which is expected based on the severity of the injuries. With an aging population, this phenomenon should severely affect resource utilization in trauma centres in the near future. Researchers should take account of both age and comorbidity in order to compare trauma populations. PMID- 16248134 TI - Noncompliance with seat-belt use in patients involved in motor vehicle collisions. AB - BACKGROUND: Seat-belt compliance in trauma patients involved in motor vehicle collisions (MVCs) appears low when compared with compliance of the general public. In this study we wished to define the relative frequency of seat-belt use in injured Canadian drivers and passengers and to determine if there are risk factors particular to seat-belt noncompliance in this cohort. METHODS: We identified trauma patients who were involved in MVCs over a 24-month period and contacted them 2-4 years after the injury by telephone to administer a standardized survey. Potential determinants of seat-belt noncompliance were compared with the occurrence of an MVC by multiple logistic regression. RESULTS: Seat-belt noncompliance in 386 MVC patients was associated with drinking and driving, youth, speeding, male sex, being a passenger, smoking, secondary roads, rural residence, low level of education, overnight driving, having no dependents, licence demerit points, previous collisions, unemployment and short journeys. There was an increase in seat-belt awareness and a decrease in self-rated driving ability after the MVC. CONCLUSIONS: Factors that indicate poor driving habits (alcohol, speeding, previous MVCs and driving offences) also predict seat-belt noncompliance. Injury prevention programs should selectively target these high risk drivers to improve seat-belt compliance and limit associated injury and consumption of health care resources. PMID- 16248135 TI - Seat-belt injuries in children involved in motor vehicle crashes. AB - BACKGROUND: The efficacy of seat belts in reducing deaths from motor vehicle crashes is well documented. A unique association of injuries has emerged in adults and children with the use of seat belts. The "seat-belt syndrome" refers to the spectrum of injuries associated with lap-belt restraints, particularly flexion-distraction injuries to the spine (Chance fractures). METHODS: We describe the injuries sustained by 8 children, including 2 sets of twins, in 3 different motor vehicle crashes. RESULTS: All children were rear seat passengers wearing lap or 3-point restraints. All had abdominal lap-belt ecchymosis and multiple abdominal injuries due to the common mechanism of seat-belt compression with hyperflexion and distraction during deceleration. Five of the children had lumbar spine fractures and 4 remained permanently paraplegic. CONCLUSIONS: These incidents illustrate the need for acute awareness of the complete spectrum of intra-abdominal and spinal injuries in restrained pediatric passengers in motor vehicle crashes and for rear seat restraints that include shoulder belts with the ability to adjust them to fit smaller passengers, including older children. PMID- 16248136 TI - Splenic syndrome in patients at high altitude with unrecognized sickle cell trait: splenectomy is often unnecessary. AB - BACKGROUND: The health risks associated with sickle cell trait are minimal in this sizable sector of the world's population, and many of these patients have no information about their sickle cell status. Splenic syndrome at high altitude is well known to be associated with sickle cell trait, and unless this complication is kept in mind these patients may be subjected to unnecessary surgery when they present with altitude-induced acute abdomen. METHODS: Four patients were admitted to the surgical ward with a similar complaint of acute severe left upper abdominal pain after arrival to the mountainous resort city of Abha, Saudi Arabia. All were subjected to splenectomy because of lack of suspicion regarding sickle cell status. RESULTS: Histologic examination of the spleen showed all patients had sickle cells in the red pulp. On further assessment all were found to have sickle cell trait with splenic infarction. In a similar study of 6 patients with known sickle cell disease who had comparable problems when they travelled to the Colorado mountains, all made an uncomplicated recovery with conservative management. CONCLUSIONS: In ethnically vulnerable patients with splenic syndrome, sickle cell trait should be ruled out before considering splenectomy. These patients could respond well to supportive management, and splenectomy would be avoided. PMID- 16248137 TI - Splenic autotransplantation for treatment of portal hypertension. AB - BACKGROUND: When total splenectomy is unavoidable it is important to preserve splenic function in some form in order to prevent the complications of asplenism. Splenic autotransplantation is a good alternative in such cases. We describe the use of splenic autotransplantation for the treatment of portal hypertension. METHODS: We carried out total splenectomy on 31 patients (21 men, 10 women), ranging in age from 21 to 68 years, with schistosomal portal hypertension. From each removed spleen, we took 20 fragments and implanted them on the greater omentum. This procedure was combined with abdominal portal-variceal disconnection. Transgastric running suture of the lower esophageal and gastric varices completed the treatment of portal hypertension. All patients underwent clinical, hematologic, immunologic and scintigraphic assessment. The results with respect to morbidity and mortality, and hematologic and immunologic findings were compared with those in 36 patients submitted to other splenic procedures. RESULTS: There was no complication related to the splenic implants and none of the patients died. Hematologic and immunologic findings were normal. Scintigraphy registered images of splenic tissue in all cases. CONCLUSION: The implantation of splenic fragments on the greater omentum seems to be a safe and useful procedure for maintaining splenic function after total splenectomy. PMID- 16248138 TI - Impact of cognitive imaging and sex differences on the development of laparoscopic suturing skills. AB - BACKGROUND: The introduction of noninvasive laparoscopic surgery has raised concerns about appropriate teaching techniques for medical students considering surgery as a specialization. The principal aim of this study was to determine the effect, between the sexes, of cognitive imaging as a teaching method in the context of learning a surgical technique. METHODS: A randomized treatment-control sample of 42 medical student volunteers was used to test the effect of cognitive imaging on performance and on traditional instructional techniques to help medical students acquire suturing skills specific to laparoscopic surgery. RESULTS: Repeated-measures analysis of variance showed no significant effect for the use of cognitive imaging (F1,40 = 0.97, p > 0.05). Males tended to perform better than females in completing tasks that required the use of visual-spatial manipulation of the instruments within a simulated laparoscopic environment (F1,40 = 5.08, p < 0.05). CONCLUSIONS: These results, which are in concordance with other research findings, indicate that females generally have lower visual spatial abilities than males. Enhanced performance for both sexes, however, increases rapidly with practice. Other than verbal one-on-one instruction, males on average rank instructional approaches that are applied and visual higher than do females. PMID- 16248139 TI - Lipofibromatous hamartoma: review of early diagnosis and treatment. AB - BACKGROUND: Lipofibromatous harmartoma (LFH) is a rare tumour that requires early diagnosis and treatment. To alert physicians to the possibility of this tumour, we review the epidemiologic and pathological characteristics, the presentation, diagnosis and treatment of LFH in the upper extremity. METHODS: We obtained data from a MEDLINE search of the English literature from 1966 to June 2003, using the search terms "lipofibromatous," "hamartomas of the nerve," "macrodactyly" and "intraneural lipoma." STUDY SELECTION: Articles that demonstrated institutional experience (epidemiologic, diagnostic and therapeutic, and complications) and those that provided general reviews were selected for inclusion. Of 501 identified articles, 42 fulfilled the review criteria. DATA EXTRACTION: Relevant clinical, diagnostic and therapeutic evaluations were extracted from the literature and synthesized. DATA SYNTHESIS: LFH is distinguished pathologically from other intraneural tumours by fibrofatty infiltration around the nerve fascicles. Typically, patients present either early with macrodactyly or later with a forearm mass lesion or symptoms consistent with a compressive neuropathy of the affected nerve. Although MRI is an important tool in the diagnosis of LFH, the diagnosis should be supported with a tissue biopsy. Treatment for patients with no neurologic deficit involves prophylactic decompression of affected nerves at all points of possible compression. Patients with macrodactyly may benefit from debulking, joint reconstruction or amputation. CONCLUSIONS: LFH differs from other benign nerve tumours with respect to clinical presentation, pathological characteristics, MRI features and management. Because LFH is extensive and inseparable from the nerve fascicles, excision is not recommended. PMID- 16248141 TI - Canadian Association of General Surgeons and American College of Surgeons evidence based reviews in surgery. 14. Preoperative fasting for adults to prevent perioperative complications. PMID- 16248140 TI - What is so odd about odds? AB - In clinical studies, the relative likelihood of an event occurring between 2 groups is often expressed as the risk ratio (RR) or the odds ratio (OR). The RR is an intuitive parameter that is relatively easy to interpret. Quantitative interpretation of an OR is much more difficult and is often incorrectly equated to that of an RR. The problem is that OR may differ substantially from RR, especially when the outcome of interest is common in the study population. This article explains and clarifies controversial issues surrounding the use and interpretation of the OR. Theoretical concepts relating to ORs are illustrated by examples from the surgical literature. By reviewing articles from 5 surgical journals over a 5-year period, we show that the OR is often presented and misinterpreted as equivalent to the RR. When the discrepancy is large, using OR uncritically as an estimate of RR will strongly bias inferences about treatment effect or cause of disease by amplifying the apparent strength of an association between an exposure and an outcome. PMID- 16248142 TI - Surgical images: soft tissue. Recurrent aneurysms. PMID- 16248143 TI - Surgical technique. A simple drain dressing. PMID- 16248144 TI - Spinal cord impingement by a displaced rib in scoliosis due to neurofibromatosis. PMID- 16248145 TI - The contralateral sentinel node. PMID- 16248146 TI - Gastrocolic fistula secondary to right gastroepiploic-coronary artery bypass. PMID- 16248147 TI - Surgical training. PMID- 16248148 TI - Procedure of choice for GIST. PMID- 16248149 TI - [The study of the mechanisms of formation of reactive oxygen species in aqueous solutions on exposure to high peak-power pulsed electromagnetic radiation of extremely high frequencies]. AB - Using the method of enhanced chemiluminescence in a peroxidase-luminol-p iodophenol system, we found the formation of reactive oxygen species (in equivalent of hydrogen peroxide concentration) in 1 mM phosphate buffer under the exposure to high peak-power pulsed electromagnetic radiation of extremely high frequencies (37 GHz, peak power 20 kW, pulse width 400 ns, repetition rate 500 Hz). The results obtained show that the formation of hydrogen peroxide in aqueous solutions under the action of electromagnetic radiation is the result of the summary influence of heat and thermoacoustic waves excited in the solutions. PMID- 16248150 TI - [The lipophilic fluorescent probe 4-dimethylaminochalcone: factors responsible for the fluorescence yield]. AB - Factors responsible for fluorescence quenching of the lipophylic fluorescent probe 4-dimethylaminochalcone in nonpolar and polar media were studied. The femtosecond dynamics of 4-dimethylaminochalcone excited state was measured using the absorption method of "excitation probing". In nonpolar hexane where the fluorescence quantum yield is very low (0.001), all excited 4 dimethylaminochalcone molecules go to the triplet state with a rate constant of 2.10(11) s(-1). At the same time, the radiation rate constant is 1000 times lower; therefore, such a fast transition to triplet is the major cause of the very small fluorescence yield. In polar acetone, the fluorescence yield is 220 times higher than in hexane. In acetone, no transitions to triplet state were detected. At the same time, a radiationless conversion to the ground state took place with a rate constant of 10(9) s(-1), which decreased the fluorescence yield to 0.22. The activation energy of the quenching processes is polarity dependent and decreases from 6 in nonpolar to 3 kcal/mol in polar media. The yield of 4 dimethylaminochalcone fluorescence varies hundreds times in media of different polarity but is a linear function of the Lippert's polarity parameter f(epsilon,n) where epsilon is the dielectric constant at low frequencies. It is supposed that polar media stabilize the "flat" conformation of the 4 dimethylaminochalcone molecule prior to its excitation, and this conformation is optimal for fluorescence. In this case, stabilization is determined only by medium polarity. PMID- 16248151 TI - [A comparative structural analysis of Fab-fragments of monoclonal rheumatoid and non-rheumatoid immunoglobulins M]. AB - The spin-labeling method was used to study the Fab- and Fab-RF-fragments of IgM and IgM-RF, respectively. The spin-label 2,2,6,6-tetramethyl-4-dichloro-sym triazinyl-aminopiperidine-1-oxyl was introduced into the peptide moiety of the proteins. The rotational correlation time t of the spin-label carrier was determined based on the temperature-viscosity dependence of the EPR spectra parameters of the spin-labeled proteins. The tau values for Fab- and Fab-RF fragments were 21 +/- 2 and 12 +/- 1 ns, respectively. The data strongly suggest that the significantly lower tau value for the Fab-RF-fragment may be due to the local structural flexibility of the fragment, which in turn may explain the peculiarities of IgM-RF as an autoantibody. PMID- 16248152 TI - [A study of the peripheral quaternary ligand binding site of cholinesterase with the use of ethidium bromide]. AB - The peripheral binding site of horse serum cholinesterase (EC 3.1.1.7) for quaternary ligands was investigated by fluorescent probing with the use of ethidium bromide. Spectral evidence for the participation of the tryptophan indole group of the peripheral site of horse serum cholinesterase in the formation of a cholinesterase complex with ethidium bromide is presented. The mechanism of cholinesterase effect on ethidium bromide fluorescence is proposed. PMID- 16248153 TI - [Myosin light chains of skeletal and cardiac muscles of ground squirrel Citillus undulatus in different periods of hibernation]. AB - The isoform composition of myosin light chains and the extent of their phosphorylation in skeletal and cardiac muscles of ground squirrel Citellus undulatus in different periods of hibernation were studied. Regulatory myosin light chains of skeletal muscles of hibernating ground squirrels were completely dephosphorylated, while 25% of these light chains in active animals were phosphorylated. During hibernation, a shift of isoform composition of essential and regulatory skeletal muscle myosin light chains toward slower isoforms was observed, which is evidenced by the data obtained on m. psoas and on the totality of all skeletal muscles. In the atrial myocardium of hibernating ground squirrels, ventricular myosin light chains 1 (up to 60%) were registered. In contrast, during arousal of ground squirrels, in ventricular myocardium the appearance of atrial myosin light chains 1 (up to 30%) was revealed. A possible role of posttranslation changes in myosin light chains and their isoform shifts in the hibernation scenario is discussed. PMID- 16248154 TI - [Sarcomeric proteins of the titin family form amyloids]. AB - It was shown for the first time that skeletal muscle sarcomeric proteins of the titin family (X-, C- and H-proteins) are able to form in vitro amyloid aggregates of different types: granular aggregates, protofibrils, helically twisted ribbons, linear fibrils, and bundles of linear fibrils. Their amyloid nature was confirmed by electron, polarization, and fluorescence microscopy and by spectral methods. As opposed to other amyloidogenic proteins, X-, C-, and H-proteins easily form amyloids under mild conditions close to physiological ones (pH, ionic strength, temperature). Like amyloid fibrils of Abeta-peptide and tau protein in Alzheimer's disease, amyloid aggregates formed by X-, C-, and H-proteins are destroyed by the antibiotic tetracycline. Thus, new proteins-precursors of amyloids and possible participants of amyloidoses in muscles were discovered. Further study of in vitro amyloidogenesis of these proteins would help to find approaches to controlling this process in organs and tissues. PMID- 16248155 TI - [Spectroscopic investigations of the structure of DNA complexes with Mn2+ in UV and IR regions]. AB - Based on the data of UV and IR spectroscopy, electronic and vibrational circular dichroism, the interaction of manganese ions with DNA was investigated. It was shown that the binding of ions to DNA proceeds in three stages depending on the manganese-to-DNA phosphates molar ratio [Mn]/[P]. At the first stage ([Mn]/[P] < or = 1), the interaction of manganese ions with DNA phosphates occurs, causing a partial screening of their negative charge and the stabilization of the double helix. At the second stage (1 < [Mn]/[P] < 6), the ions interact with both the phosphates and the nitrogen bases of DNA. At this stage, it is possible for the manganese ion to coordinate simultaneously to the oxygen atom of the phosphate and the neighbouring base of DNA. At a higher [Mn]/[P] ratio, the destabilization of the double helix begins, and partial breakage of the hydrogen bonds between the nitrogen bases occurs. PMID- 16248156 TI - [Interactions of caffeine with DNA double helix fragments. Molecular mechanics simulation]. AB - Calculations of the energy of interaction between the caffeine molecule and DNA double helix fragment of four complementary pairs have been performed by the molecular mechanics method. The calculations demonstrate the existence of energy minima corresponding to the caffeine molecule position in both wide and narrow grooves. Each of three proton acceptor atoms of caffeine is able to form hydrogen bond with each of three amino groups of DNA bases. The interactions of caffeine with both hydrogen bonded nucleotide and other nucleotides of the two strands contribute considerably to the total energy. The substantial contribution of interactions of caffeine with other than H-bonded nucleotides results in a rather close packing of atom groups in possible DNA-caffeine complexes. The mechanisms of influence of caffeine on interactions of DNA with other biologically active compounds are discussed. PMID- 16248157 TI - [The formation of nanoconstructions based on double-stranded DNA molecules]. AB - The properties of nanoconstructions formed by double-stranded DNA molecules fixed in the structure of their liquid crystalline dispersions and cross-linked by nanobridges were described. The dependence of the optical properties of the nanostructures on various factors (DNA concentration, nanobridge components, etc.) was examined. PMID- 16248158 TI - [A new approach to the problem of oxygen formation in photosynthesis]. AB - A detailed quantum-mechanical analysis of the model of water oxidizing complex, based on recent X-ray data on the structure of PSII, was made. A mechanism of water oxidation was suggested and explained for the first time. The role of three manganese atoms that are not involved directly in water oxidation, the role of the cubic structure of the complex, and the necessity of the presence of calcium and chlorine atoms during water oxidation are discussed. Theoretical computations of the energies of the complex in each S-state were made. PMID- 16248159 TI - [On the possibilities of diagnosis of the state of the phytoplankton photosynthetic apparatus by the method of nonlinear laser fluorimetry]. AB - The possibilities of diagnosis of the state of the phytoplankton photosynthetic apparatus by the method of nonlinear laser fluorimetry (saturation fluorimetry), which can be realized in remote sensing mode, were investigated. A procedure for the determination of the nonsaturated fluorescence parameter phi0, which is proportional to the concentration of chlorophyll a molecules, and the parameter A, which is a product of three photophysical parameters of chlorophyll a molecule in native chloroplast, was elaborated. Laboratory experiments with the axenic culture of eurihaline Thalassiosira weissflogii showed that the parameter A depends on the state of the photosynthetic apparatus of the alga, which was varied by either DCMU treatment or exposure to actinic light. PMID- 16248160 TI - [Deterministic chaos in the dynamics of biomembrane ion channel current]. AB - Within the framework of the theory of deterministic chaos, a nonlinear first order differential equation with delay and relaxation with periodic influence on channel current for the parameter of order (deviation of channel current from the equilibrium value) was obtained. The numerical solutions of the equation indicate a chaotic dynamics of the order parameter and conformation potential of the channel protein with positive Lyapunov indices. By integration in the time interval between the "jumps" of ions through energy barriers of the channel protein, a mapping was obtained that also results in chaotic solutions realized in experiments. Basic kinetic characteristics of ionic channels for the mapping were obtained: the probability for the channel to be in the open state, P0, and the mean duration of a pack of current pulses depending on controlling parameters. Algorithms for constructing bifurcation diagrams with the transition to chaos and for determining Lyapunov indices and Kholmogorov entropy, pulsation spectra, and other parameters of chaotic dymanics were developed. PMID- 16248161 TI - [The role of lipid peroxidation products in the effect of He-Ne laser on human blood leukocytes]. AB - The role of lipid peroxidation products formed in membranes of human blood leukocytes after irradiation with He-Ne laser was studied. It was found that low intensity laser irradiation (0.3-1.6 J/cm2) leads to both cell activation and an increase in the content of lipid peroxidation products. The intensity of lipid peroxidation was analyzed by estimating the amount of TBA reactive products and lipid diene conjugates. Irradiation in the presence of an exogenous photosensitizer (protoporphyrin IX) enhanced the phenomena observed. The use of antioxidants (tocopherol and ionol) completely eliminated the laser-induced effects (changes in leukocyte activity and accumulation of lipid peroxidation products). These results can be explained by the fact that laser irradiation leads to the activation of lipid peroxidation in leukocyte membranes, which in turn enhances the response of cells to the stimulus (priming). PMID- 16248162 TI - [A soft poration of planar bilayer lipid membranes from dipalmitoylphosphatidylcholine at the temperature of the phase transition from the liquid crystalline to the gel state]. AB - A method of soft poration of lipid bilayer was suggested, which is based on the structural rearrangement of lipid bilayer formed from disaturated phospholipids on the phase transition from liquid crystalline state to the gel. As opposed to the widely used method of electropbration, this method allows one to obtain a lipid pore population without application of high electric field. In the case of soft poration, the electric field does not exceed the physiological level of 10 100 mV. It was shown that, in planar bilayer lipid membranes formed from dipalmitoylphosphatidylcholine in water solution of 1 M LiCl, there appear up to 10 lipid pores in 1 min per 1 mm of membrane surface with an average conductivity of a pore of 31 +/- 13 nS. The average pore radius estimated using soluble polyethylene glycols ranged between 1.05-1.63 nm. Monovalent cation conductivity of a single lipid pore on soft poration was shown to decrease in the order Li+ > or = Na+ > K+ = Rb+ > or = Cs+. This order coincides with that observed by Marra and Israilashvili for dipalmitoylphosphatidylcholine-water interbilayer where the repulsive hydration force contribution is significant. PMID- 16248163 TI - [A study of water autodiffusion in model biological membranes, oriented lipid bilayers]. AB - The autodiffusion of water in a multibilayer structure formed by dipalmitoyl phosphatidylcholine and oriented on glass plates was studied by the method of NMR with magnetic field pulse gradient. It was shown that water molecules occur in several states differing in the degree of interaction with lipid molecules. A spectrum of the coefficients of water autodiffusion in a direction transversal to bilayers was found. The use of samples with different distances between the plates and an analysis of the dependence of the mode of diffuse decay of spin echo on diffusion time and the orientation of the sample, as well as measurements at temperatures above and below the gel-liquid crystal phase transition in cholesterol-containing samples enabled one to discriminate the diffuse decay component responsible for the transbilayer movement of water. The coefficient of bilayer permeability was estimated using the Tanner model. It was shown that the formation of mechanical defects ("cracks") in plane oriented bilayers is the most probable reason for the presence of the water component with the relatively high coefficient of diffusion. PMID- 16248165 TI - [Discharge frequency and excitability of human firing motoneurone]. AB - The excitability of firing motoneurones activated by voluntary contraction of the flexor carpi ulnaris or tibialis anterior was tested by single excitatory Ia afferent volleys. In order to estimate the stimulation effects, the peri-stimulus time histograms of single motoneurones were plotted, and the firing indices were calculated. It was shown that the firing-frequency effect was absent within the range of 4-14 imp/s during testing by low-intensity excitatory volley. At higher intensity of afferent volley, the excitability increased at a low firing rate. It is suggested that the characteristics of the interspike-interval excitability trajectories underlie these relations. These findings made it possible to explicate some conflicting literature data which were usually reported without taking the afferent volley intensity effect into account. The mechanisms controlling the firing motoneurone excitability and possible trajectories of interspike-interval membrane potential in human motoneurones are discussed. PMID- 16248164 TI - [The interaction of dynamic speckles with suspensions of gram-negative cells]. AB - The role of temporal coherence at photodynamic action of light on living cells was investigated. A mathematical model describing the interaction of low-coherent speckles with bacterial cells was suggested and its parameters were determined based on experimental data. The interrelation between the life time of dynamic optical speckles and the degree of photodestruction of illuminated cells was established by computer simulations. The conditions were determined under which the photoinactivation of cells of the vaccine strain of the causative agent of F. tularensis 15 NIIEG occurs. PMID- 16248166 TI - [A study of the aggregation of human red blood cells induced by picric acid]. AB - The effect of picric acid on the aggregation of human erythrocytes was studied. It was shown that the addition of picric acid to a suspension of washed erythrocytes leads to a decrease in pH of medium to 1.5-2 and the formation of echinocytes. Stirring the suspension of echinocytes at low pH values results in a strong aggregation of cells. Increasing the pH value to 7.4 leads to a desaggregation of echinocytes. It was found that picric acid does not induce the aggregation of cells fixed by glutaraldehyde. A substantial decrease in the aggegation of spheric erythrocytes obtained after heating the cells at 50 degrees C was observed. PMID- 16248167 TI - [A possible molecular mechanism of the narcotic action of noble gases]. AB - A molecular mechanism of the narcotic action of noble gases is suggested, which is based on the fact that noble gas atoms change the orientation of water molecules absorbed on the surface of axon membrane. The resulting change in the transmembrane potential deteriorates the propagation of nerve pulse. PMID- 16248169 TI - [The investigation of the synchronization between rhythms in the human cardiovascular system from time series of R-R-intervals]. AB - We demonstrated that, from the sequence of R-R intervals, it is possible to calculate the instantaneous phases and instantaneous frequencies of the main rhythmic processes governing the cardiovascular dynamics in humans, namely, the main heart rhythm, respiration, and the process of slow regulation of blood pressure with basic frequency close to 0.1 Hz. For the cases of spontaneous respiration and paced respiration with a fixed frequency, the synchronization between the rhythms of the cardiovascular system was investigated based on the analysis of only the time series of R-R intervals. It is shown that the main heart rhythm and the rhythm of low-frequency regulation of blood pressure can be synchronized with respiration. PMID- 16248168 TI - [The effect of sphyngolipids on the mechanical properties of epidermis and its permeability for water]. AB - The effect of water, glycerol, Lipotec(s.a), a hydrophobic sphyngolipid complex, and a liposome water emulsion (prepared from pig brain glycosphyngolipids) on the skin elasticity, evaluated from its resonance frequency, was investigated. It was shown that moistening of skin with water leads to swelling of epidermis cells, which is accompanied by an increase in cellular membrane tension and elasticity growth. This shows up in a statistically authentic (alpha = 0.0025, T-test) growth of skin resonance frequency (on the average by 48.5 +/- 13% in 3 min). Skin moistening by a glycosphyngolipid liposome water emulsion causes a more intensive swelling of epidermis (skin resonance frequency in three minutes increases on the average by 75.8 +/- 22.1%, alpha = 0.0059). No swelling of epidermis was observed if skin was moistened by glycerol or by hydrophobic sphyngolipid complex Lipotec(s.a). It was concluded that pig brain glycosphyngolipid molecules having volumetric negatively charged polar heads are built into lamellar structures of the skin lipid barrier and increase its hydrophilicity and permeability for water. PMID- 16248170 TI - [Modeling the interactions of pacemakers of sigma rhythm spindles]. AB - An approach to modeling "sleeping" spindles (waxing and waning of characters patterns cycles 12-18 H) of electrical activity of mammalian brain was proposed, which is based on the idea of the pacemaker nature of the sigma rhythm. The spindles are assumed to result from the rhythmic interaction of cortical neurons and medial thalamus cells. A system of oscillatory links operating in the mode of small vacillations is taken as an analog of this interaction. A deepening of slow wave sleep is considered to result from an increase in the density of interactions and the number of oscillatory links, which is associated with differences in the adaptation of organisms of high and low anxiety to environmental changes. PMID- 16248171 TI - [Evolution of rotifer population dynamics in a heterogeneous habitat: mathematical modeling]. AB - We present the results of mathematical modeling of a rotifer species inhabiting two coupled habitats with different environmental conditions. We use the modified Consensus model and show that the exchange between the habitats can lead to chaotization of originally regular plankton dynamics and synchronization of plankton biomass oscillations. As a result, the invasion of a chaotic regime takes place. PMID- 16248172 TI - [The significance of modular design in the investigation of processes of system self-organization]. AB - A model of the process of determined system self-organization based on system forming modular water structures is proposed. The arrangement and symmetry of these structures, described by the symmetry groups entanglement, matches the basic principles of system self-organization: "system of systems", "recognition", "all-or-none". Crystallography modular generalization engulfs all stable forms of condensed state, including the bound water structures--matrices for the self organization of biological systems. The bound water structures, besides being matrices, accomplish the metric selection of other structural components of biological systems capable of self-organization into a whole system by creating numerous directed weak bonds among them. PMID- 16248173 TI - [Could life evolve by random mutations?]. AB - The problem of the rate and mechanisms of evolution was considered. It was shown that organisms could not be formed by random mutations during real times of about one million years. It was concluded that deterministic models are necessary for the description of evolution. PMID- 16248174 TI - Runoff modelling using radar data and flow measurements in a stochastic state space approach. AB - In urban drainage the estimation of runoff with the help of models is a complex task. This is in part due to the fact that rainfall, the most important input to urban drainage modelling, is highly uncertain. Added to the uncertainty of rainfall is the complexity of performing accurate flow measurements. In terms of deterministic modelling techniques these are needed for calibration and evaluation of the applied model. Therefore, the uncertainties of rainfall and flow measurements have a severe impact on the model parameters and results. To overcome these problems a new methodology has been developed which is based on simple rain plane and runoff models that are incorporated into a stochastic state space model approach. The state estimation is done by using the extended Kalman filter in combination with a maximum likelihood criterion and an off-line optimization routine. This paper presents the results of this new methodology with respect to the combined consideration of uncertainties in distributed rainfall derived from radar data and uncertainties in measured flows in an urban catchment within the Emscher river basin, Germany. PMID- 16248175 TI - Modelling non-structural best management practices--focus on reductions in stormwater pollution. AB - This paper describes a modelling approach for evaluating the efficiency of different non-structural best management practices for stormwater management. A scenario with a set of source reduction practices was simulated using the substance flow model SEWSYS for an urban catchment in the city of Goteborg, Sweden. The scenario is based on a hypothetical control program that includes prevention, education and regulations. The simulation shows relatively high reductions of copper and PAH, 77% and 50%, respectively. The reduction in copper is mainly due to less copper roof corrosion and brake wear, while reduced road wear has the greatest effect for PAH. An important result from this study is that the nonstructural BMPs applied did not give a sufficient reduction in pollution to meet the desirable environmental quality criteria. To meet these criteria, additional BMPs must be implemented, preferably a combination of both non structural and structural measures. PMID- 16248176 TI - Ecotechnology: basis of a new immission concept in water pollution control. AB - Beyond the traditional load reduction also an ecosystem-internal mechanism can be used to minimise the effects of water pollution. The control of the internal mechanisms is achieved through the optimisation of the ecosystem structure. This ecotechnology principle is based on the idea to reduce as much as possible the gap between the current (suboptimal) structural status and the optimum structure by intentional manipulations. The spectrum of such manipulations is very broad. A few examples are demonstrated. They comprise physical (e.g. stream morphology), chemical (e.g. enhancing the redox potential at the sediment-water interface) and biological (e.g. enhancing stocks of predatory fishes) control measures. It can be supposed that a new immission concept including the ecotechnology principle could be much more adequate to the demand of modern water pollution control than the traditional emission and imission concepts. PMID- 16248177 TI - Surface models for coupled modelling of runoff and sewer flow in urban areas. AB - Traditional methods fail for the purpose of simulating the complete flow process in urban areas as a consequence of heavy rainfall and as required by the European Standard EN-752 since the bi-directional coupling between sewer and surface is not properly handled. The new methodology, developed in the EUREKA-project RisUrSim, solves this problem by carrying out the runoff on the basis of shallow water equations solved on high-resolution surface grids. Exchange nodes between the sewer and the surface, like inlets and manholes, are located in the computational grid and water leaving the sewer in case of surcharge is further distributed on the surface. Dense topographical information is needed to build a model suitable for hydrodynamic runoff calculations; in urban areas, in addition, many line-shaped elements like houses, curbs, etc. guide the runoff of water and require polygonal input. Airborne data collection methods offer a great chance to economically gather densely sampled input data. PMID- 16248178 TI - Impact of dimension uncertainty and model calibration on sewer system assessment. AB - Assessments of sewer performance are usually based on a single computation of CSO (combined sewer overflow) volumes using a time series of rainfall as system loads. A shortcoming of this method is that uncertainties in knowledge of sewer system dimensions are not taken into account. Moreover, sewer models are rarely calibrated. This paper presents the impacts of database errors and model calibration on return periods of calculated CSO volumes. The impact of uncertainties is illustrated with two examples. Variability of calculated CSO volumes is estimated using Monte Carlo simulations. The results show that calculated CSO volumes vary considerably due to database errors, especially uncertain dimensions of the catchment area. Furthermore, event-based calibration of a sewer model does not result in more reliable predictions because the calibrated parameters have low portability. However, it enables removal of database errors harmonising model predictions and 'reality'. PMID- 16248179 TI - Automatic calibration of urban drainage model using a novel multi-objective genetic algorithm. AB - In order to successfully calibrate an urban drainage model, multiple calibration criteria should be considered. This raises the issue of adopting a method for comparing different solutions (parameter sets) according to a set of objectives. Amongst the global optimization techniques that have blossomed in recent years, Multi Objective Genetic Algorithms (MOGA) have proved effective in numerous engineering applications, including sewer network modelling. Most of the techniques rely on the condition of Pareto efficiency to compare different solutions. However, as the number of criteria increases, the ratio of Pareto optimal to feasible solutions increases as well. The pitfalls are twofold: the efficiency of the genetic algorithm search worsens and decision makers are presented with an overwhelming number of equally optimal solutions. This paper proposes a new MOGA, the Preference Ordering Genetic Algorithm, which alleviates the drawbacks of conventional Pareto-based methods. The efficacy of the algorithm is demonstrated on the calibration of a physically-based, distributed sewer network model and the results are compared with those obtained by NSGA-II, a widely used MOGA. PMID- 16248180 TI - Model description of storage and infiltration functions of infiltration facilities for urban runoff analysis by a distributed model. AB - Although there have been simulation researches focusing on reduction of stormwater peak flow by introduced infiltration facilities, model simulation of dynamic runoff behavior is still limited for frequently occurring rainfall events with weak intensity. Therefore, dynamic simulation was carried out in two urban drainages with infiltration facilities incorporated with a distributed model using two methods for describing functions of infiltration facilities. A method adjusting effective rainfall model gave poor simulation of runoff behavior in light rainfalls. Another method considering dynamic change of storage capacity as well as infiltration rate gave satisfactory estimation of the runoff in both drainages. In addition, assumption of facility clogging improved the agreement between measured and simulated hydrographs in small and medium-sized rainfall. Therefore, the proposed method might be useful for quantifying the secondary effects of the infiltration facilities on groundwater recharge and urban non point pollutant trapping as well as runoff reduction. PMID- 16248181 TI - Stormwater quality models: sensitivity to calibration data. AB - Stormwater quality modelling is a useful tool in sewer systems management. Available models range from simple to detailed complex ones. The models need local data to be calibrated. In practice, calibration data are rather lacking. Only few measured events are commonly used. In this paper, the effect of the number and the variability of calibration data on models of various levels of complexity are investigated. The study is carried out on "Le Marais" catchment for suspended solids where 40 reliable measured events and good knowledge of the sewer system are available. The method used is based on resampling subsets of measured events among the 40 available ones. Three types of models were calibrated using subsets of events of different sizes and characteristics resampled among the 40 available ones. For each calibration, the model was validated against the remaining events to stand upon the quality of the model. It was found that the models are quite sensitive to calibration data, a problem neglected in practical studies. The use of more complex models does not necessarily improve modelling results since more problems and error sources are to be expected. The findings are specific to "Le Marais" catchment and the models used. PMID- 16248182 TI - A model for the movement of large solids in small sewers. AB - An extensive series of experiments has been carried out to investigate the movement mechanisms and behaviour of large solids in small sewers. This paper describes the development, calibration and verification of a model (SOLID) based on data obtained from the experimental rig. It is used to predict solid movement with respect to 'limiting solid transport distance'. Key model parameters are the coefficients of static and dynamic friction, the shape factor of amorphous solids and the flow bypass coefficient. The model is shown to successfully represent the movement of a large solid down a small pipe, where the solid is moving as a sliding, leaking dam, particularly the first 'hop'. Limitations of the model include a limited facility to well represent multiple hops and the need for closely spaced computational nodes leading to small time steps, and long run times. PMID- 16248183 TI - Uncertainty in predicting riverbed erosion caused by urban stormwater discharge. AB - Ecologically based criteria require an integrated modeling approach. Due to the complexity of the system, the stochastic nature of loads, and the model abstractions, many uncertainties are involved. In this study, a simple integrated model is applied, which Swiss engineers employ to assess the impact of urban stormwater discharges on riverbed stability. In the course of a case study, an uncertainty analysis is carried out focusing on parameter uncertainties. The underlying context of the uncertainties is evaluated, and a variance-based sensitivity analysis is presented estimating the local uncertainty contribution of each parameter. The results reveal that the largest contributions stem from the model components describing the natural system. An experimental design is proposed that manages to reduce the output uncertainty significantly. Finally, we discuss the benefits of following the proposed procedure. PMID- 16248184 TI - Model based hydropower gate operation for mitigation of CSO impacts by means of river base flow increase. AB - In river stretches being subjected to flow regulation, usually for the purpose of energy production (e.g. Hydropower) or flood protection (river barrage), a special measure can be taken against the effect of combined sewer overflows (CSOs). The basic idea is the temporal increase of the river base flow (during storm weather) as an in-stream measure for mitigation of CSO spilling. The focus is the mitigation of the negative effect of acute pollution of substances. The measure developed can be seen as an application of the classic real time control (RTC) concept onto the river system. Upstream gate operation is to be based on real time monitoring and forecasting of precipitation. The main objective is the development of a model based predictive control system for the gate operation, by modelling of the overall wastewater system (incl. the receiving water). The main emphasis is put on the operational strategy and the appropriate short-term forecast of spilling events. The potential of the measure is tested for the application of the operational strategy and its ecological and economic feasibility. The implementation of such an in-stream measure into the hydropower's operational scheme is unique. Advantages are (a) the additional in stream dilution of acute pollutants entering the receiving water and (b) the resulting minimization of the required CSO storage volume. PMID- 16248185 TI - An integrated modelling concept for immission-based management of sewer system, wastewater treatment plant and river. AB - Today's planning standards deal with the individual urban drainage components (sewer system, wastewater treatment plant and receiving water) separately, i.e. they are often designed and operated as single components. As opposed to this, an integral handling considers the drainage components jointly. This novel approach allows a holistic and more sustainable planning of urban drainage systems. This paper presents an integrated modelling concept. The aim is to analyse fluxes through the total wastewater system and to integrate pollution-based control in the upstream direction, that is, e.g., managing the combined water retention tanks as a function of state variables in the WWTP or the receiving water. All models of the different subsystems are based on the Activated Sludge Model (ASM) concept of IWA, including River Water Quality Model No. 1 (RWOM). Simulations can be done in truly parallel mode using the simulation environment SIMBA. The integrated modelling concept is applied to the river Dhuenn and the urban wastewater system of the municipality of Odenthal (Germany). An optimised operation of the system using RTC proves to be a very effective measure. PMID- 16248186 TI - Modelling of different measures for improving removal in a stormwater pond. AB - The effect of retrofitting an existing pond on removal efficiency and hydraulic performance was modelled using the commercial software Mike21 and compartmental modelling. The Mike21 model had previously been calibrated on the studied pond. Installation of baffles, the addition of culverts under a causeway and removal of an existing island were all studied as possible improvement measures in the pond. The subsequent effect on hydraulic performance and removal of suspended solids was then evaluated. Copper, cadmium, BOD, nitrogen and phosphorus removal were also investigated for that specific improvement measure showing the best results. Outcomes of this study reveal that all measures increase the removal efficiency of suspended solids. The hydraulic efficiency is improved for all cases, except for the case where the island is removed. Compartmental modelling was also used to evaluate hydraulic performance and facilitated a better understanding of the way each of the different measures affected the flow pattern and performance. It was concluded that the installation of baffles is the best of the studied measures resulting in a reduction in the annual load on the receiving lake by approximately 8,000 kg of suspended solids (25% reduction of the annual load), 2 kg of copper (10% reduction of the annual load) and 600 kg of BOD (10% reduction of the annual load). PMID- 16248187 TI - Modeling stochastic load variations in sewer systems. AB - In wastewater treatment and environmental risk assessments increasing attention is paid to the fate of micropollutants. These are time-consuming, expensive and difficult to detect and quantify. If a substance's load or concentration is subject to high dynamic fluctuations, it is demanding to take representative samples, especially when the "variation" is unknown. Therefore, we developed a concept to model stochastic load variations in sewer systems. We gathered readily available information from existing databases (population and consumption data) and combined it with the characteristics of household activities and appliances. We succeeded in predicting realistic short-term variations of benzotriazole (contained in dishwasher detergents) and validated them with a high-frequency measuring campaign. Benzotriazole stands as an example for other household chemicals, which cannot be measured so easily. All required information used within this case study is also available for other substances and catchments. This allows the forecast of stochastic load variations for many chemical compounds of interest. It helps to plan measuring campaigns, to estimate discharged loads from combined sewer overflows and to have a characteristic input for modeling purposes. PMID- 16248188 TI - Dispersion coefficients of sewers from tracer experiments. AB - In this paper, 60 tracer experiments in 37 different sewer reaches have been analyzed for longitudinal dispersion under dry weather flow conditions. It was found that dispersion coefficients of sewers are two to three orders of magnitude smaller than those measured in rivers and do not differ much from system to system. Suitable equations were identified to predict reasonable dispersion coefficients in sewer reaches with uniform geometry and stable flow conditions. For engineering applications that require a high degree of accuracy the performance of tracer measurements is recommended. PMID- 16248189 TI - Bayesian analysis for erosion modelling of sediments in combined sewer systems. AB - Previous research has confirmed that the sediments at the bed of combined sewer systems are the main source of particulate and organic pollution during rain events contributing to combined sewer overflows. However, existing urban stormwater models utilize inappropriate sediment transport formulas initially developed from alluvial hydrodynamics. Recently, a model has been formulated and profoundly assessed based on laboratory experiments to simulate the erosion of sediments in sewer pipes taking into account the increase in strength with depth in the weak layer of deposits. In order to objectively evaluate this model, this paper presents a Bayesian analysis of the model using field data collected in sewer pipes in Paris under known hydraulic conditions. The test has been performed using a MCMC sampling method for calibration and uncertainty assessment. Results demonstrate the capacity of the model to reproduce erosion as a direct response to the increase in bed shear stress. This is due to the model description of the erosional strength in the deposits and to the shape of the measured bed shear stress. However, large uncertainties in some of the model parameters suggest that the model could be over-parameterised and necessitates a large amount of informative data for its calibration. PMID- 16248191 TI - Modelling of sedimentation and remobilization in in-line storage sewers for stormwater treatment. AB - A special arrangement of combined sewer overflow tanks is the in-line storage sewer with downstream discharge (ISS-down). This layout has the advantage that, besides the sewer system, no other structures are required for stormwater treatment. The verification of the efficiency with respect to the processes of sedimentation and remobilization of sediment within the in-line storage sewer with downstream discharge is carried out in a combination of a field and a pilot plant study. The model study was carried out using a pilot plant model scaled 1:13. The following is intended to present some results of the pilot plant study and the mathematical empirical modelling of the sedimentation and remobilization process. PMID- 16248190 TI - Improved formulations for rapid erosion of diverse solids in combined sewers. AB - For more than a decade, research carried out in Scotland has investigated the movement of sediment in sewers and the associated pollutant release. Pollution by discharges from combined sewer overflows can adversely affect watercourses, particularly those in urban areas. Solids and dissolved contaminants, many derived from in-sewer deposits during a storm event, can be especially significant. This phenomenon can occur during events known as 'foul flushes'. In combined sewers these typically occur in the initial period of storm flows, when the concentration of suspended sediments and other pollutants is significantly higher than at other times. It has become apparent that much of the suspended load originates from solids eroded from the bed. The 'near bed solids' which are re-entrained into the flow, together with solids eroded from the bulk bed, account for large changes in the suspended sediment concentration under time varying flow conditions. This paper describes some of the methods employed to investigate the solids eroding in combined sewers during peak flow events. The work examined the potential for sediment re-suspension under high flow conditions both in the laboratory and in the field. PMID- 16248192 TI - A long-term suspended solids runoff simulation in a highway drainage system. AB - A long-term monitoring investigation was carried out in a highway drainage system in Winterthur, Switzerland. Several runoff quality and quantity simulations were carried out using the distributed model 'InfoWorks-CS'. Serial rainfall monitoring data was used for investigation of SS runoff behavior from the highway. Under continuous rainfall conditions, the quantity simulation showed a good agreement with the measured hydrograph. However, in some cases where rainfall was not continuous, overestimation of the peak height was found at the later stage after the end of the rainfall. It was believed that the initial/depression loss on the road surface was recovered during the halting period of rainfall. The consideration of regenerated depression loss significantly improved runoff simulation results in the on-and-off type rainfall events. A single event quality simulation underestimated the SS load in light rainfall events. One of the reasons was possibly inadequate consideration of pipe sediment conditions. A long term simulation was carried out to establish the initial condition of surface and pipe sediment for the target event. The newly simulated pollutograph gave a good agreement with the measured one. It revealed that it was essential to consider appropriateness of the initial condition of pipe sediment as well as surface sediment. PMID- 16248193 TI - Dynamics of rain-induced pollutographs of solubles in sewers. AB - When looking at acute receiving water impacts due to combined sewer overflows the characteristics of the background diurnal sewage flux variation may influence the peak loads from combined sewer overflows (CSO) and wastewater treatment plant (WWTP) effluent significantly. In this paper, effects on the dynamic compounds transported in the sewer, on CSO discharges and WWTP loading are evaluated by means of hydrodynamic simulations. The simulations are based on different scenarios for diurnal dry-weather flow variations induced by different infiltration rates. PMID- 16248194 TI - Integrated real-time control for a sequencing batch reactor plant and a combined sewer system. AB - Integrated real-time control (RTC) concepts, which are trying to operate drainage systems and WWTPs depending on the current capacities of both systems, are becoming more and more important. While almost all publications in this field have been concentrating on continuous flow systems, this paper will present a project which has been initiated to realise an integrated RTC strategy for a Sequencing Batch Reactor (SBR) plant in simulation as well as in full-scale. The results of the simulation are that SBR plants can handle high hydraulic loads. The cost-benefit analysis shows that an integrated operation is reasonable concerning environmental and economic aspects. In order to verify the simulation results, full-scale operation has been started in January 2004. The first results seem to confirm the results of the simulation study. PMID- 16248195 TI - Development and implementation of a real-time control strategy for the sewer system of the city of Vienna. AB - The paper describes the realization of a real-time control for the Vienna sewer system. The project is scheduled for completion for 2004. The 3.5 year project comprises all planning stages starting with the recording of data up to the planning of measuring and controlling units. The concrete steps of the planning stages are explained. A measuring system including 25 rainfall measurements, 40 flow measurements and 20 water level measurements is implemented as an online system. This measuring system is designed to achieve two objectives, on the one hand the real-time control and on the other hand the calibration of the model that is used for the hydrodynamic sewer system simulation. The approx. 53,000 pipes have served to generate a coarse network of no more than approx. 2600 pipes. The area data were derived with high accuracy from available aerial photograph interpretations. With simulation runs of a rule-based control software the system operation was examined. A self-learning system will improve the rule basis. A forecasting model that uses weather observation radar will additionally influence the controlling decisions. The findings from the investigations are immediately considered in the planning of measuring and control units. The simulated results for the first phase of implementation, which demonstrate the benefit of RTC for the Vienna sewer system, are explained. PMID- 16248196 TI - Integrated operation of sewer system and WWTP by simulation-based control of the WWTP inflow. AB - In recent years numerical modelling became a standard procedure to optimise urban wastewater systems design and operation by integration. For dynamic control of the wastewater teatment plant (WWTP) inflow, a model-based predictive concept is introduced aiming at improving the receiving water quality. An on-line simulator running parallel to the real WWTP operation reflects the actual state of operation and provides this model information to a prognosis tool which determines the best option for the WWTP inflow. The investigations showed that it is possible to reduce the NH4-N peak concentrations in the receiving water by dynamic WWTP inflow control based on predictive scenario analysis. PMID- 16248198 TI - Integrated modelling for the evaluation of infiltration effects. AB - The objective of the present study is the estimation of the potential benefits of sewer pipe rehabilitation for the performance of the drainage system and the wastewater treatment plant (WWTP) as well as for the receiving water quality. The relation of sewer system status and the infiltration rate is assessed based on statistical analysis of 470 km of CCTV (Closed Circuit Television) inspected sewers of the city of Dresden. The potential reduction of infiltration rates and the consequent performance improvements of the urban wastewater system are simulated as a function of rehabilitation activities in the network. The integrated model is applied to an artificial system with input from a real sewer network. In this paper, the general design of the integrated model and its data requirements are presented. For an exemplary study, the consequences of the simulations are discussed with respect to the prioritisation of rehabilitation activities in the network. PMID- 16248197 TI - Monitoring of carbamazepine concentrations in wastewater and groundwater to quantify sewer leakage. AB - Monitoring of carbamazepine concentrations in wastewater and groundwater enables us to identify and quantify sewer exfiltration. The antiepileptic drug carbamazepine is hardly removed in wastewater treatment plants and not or just slightly attenuated during bank infiltration and subsoil flow. Concentrations in wastewater are generally 1000 times higher than the limit of quantification. In contrast to . many other wastewater tracers carbamazepine is discharged to the environment only via domestic wastewater. The results from this study carried out in Linz, Austria indicate an average exfiltration rate of 1%, expressed as percentage of the dry weather flow that is lost to the groundwater on the city wide scale. This rate is lower than sewage losses reported in most other studies which attempted to quantify exfiltration on the basis of groundwater pollution. However, it was also possible to identify one area with significantly higher sewage losses. This method seems to be very suitable for the verification of leakage models used to assess sewer exfiltration on a regional scale. PMID- 16248200 TI - Modelling of flushing waves for optimising cleaning operations. AB - In the framework of a German field and numerical study the possibilities of an optimal cleaning by flushing of a combined sewer with storage capacity and bottom end overflow were investigated. The objective of this combined investigation was to find out the boundaries for the flushing operation to prevent wave reflections and impoundages at the throttle by guaranteeing the necessary shear stress for remobilization. PMID- 16248199 TI - Application of a leakage model to assess exfiltration from sewers. AB - The exfiltration of wastewater from sewer systems in urban areas causes a deterioration of soil and possibly groundwater quality. Beside the simulation of transport and degradation processes in the unsaturated zone and in the aquifer the analysis of the potential impact requires the estimation of quantity and temporal variation of wastewater exfiltration. Exfiltration can be assessed by the application of a leakage model. The hydrological approach was originally developed to simulate the interactions between the groundwater and surface water, it was adapted to allow for modelling of interactions between groundwater and sewer system. In order to approximate the exfiltration specific model parameters infiltration specific parameters were used as a basis. Scenario analysis of the exfiltration in the City of Dresden from 1997 to 1999 and during the flood event in August 2002 shows the variation and the extent of exfiltration rates. PMID- 16248201 TI - Development of a distributed hydrological model based on urban databanks- production processes of URBS. AB - The objective of this study is to present a distributed hydrological model especially dedicated to urban catchments, and able to represent hydrological processes usually neglected in urban modelling, such as evapotranspiration, infiltration in roads, or direct infiltration of soil water in sewers. This model, called URBS (as Urban Runoff Branching Structure) is distributed considering the spatial variability of land use which is well known thanks to urban databanks managed by GIS. The production function is detailed at each cadastral parcel scale, and the runoff produced is routed by a simple transfer function. The estimation of the input parameters of the model is mostly based on physical considerations, and the model is applied on a suburban catchment in Nantes (France) in order to evaluate the interest of the distribution of the hydrological variables. PMID- 16248202 TI - ArcEGMO-URBAN--hydrological model for point sources in river basins. AB - The new model ArcEGMO-URBAN aims at deterministic and spatiotemporal modelling of water, nitrogen and phosphorus fluxes from all urbanised areas of a river basin considering all potential sources. Pollution loads are calculated for discrete urban patches and balanced on the level of hydrological subbasins. Modelling results can be defined by the user of any level of spatial and/or temporal aggregation, e.g. matter balances for river basins or river sections and years or months, respectively. To process spatial data, a Geographic Information System is linked to the model. Information on urban land use and general characteristics of river basins is based on digital coverages, partly generated from remote-sensing data. Moreover, statistical data, e.g. on population, sewer systems, wastewater treatment plants etc. are included. Stormwater runoff from impervious surfaces is calculated as one input to the sewer network. Wastewater is considered with its main sewer system, pumping stations and treatment plants. Finally, the discharge is balanced for discrete river sections. Modelling results attest ArcEGMO-URBAN its ability to realistically quantify matter fluxes and major pollution sources as well as their seasonal variation. This makes the model an applicable tool for the analysis of scenarios with e.g. varying population distribution or climatic and technological conditions. PMID- 16248203 TI - Assessment of urban flooding by dual drainage simulation model RisUrSim. AB - The European research project in the EUREKA framework, RisUrSim is presented with its overall objective to develop an integrated planning tool to allow cost effective management for urban drainage systems. The project consortium consisted of industrial mathematics and water engineering research institutes, municipal drainage works as well as an insurance company. The paper relates to the regulatory background of European Standard EN 752 and the need of a more detailed methodology to simulate urban flooding. The analysis of urban flooding caused by surcharged sewers in urban drainage systems leads to the necessity of a dual drainage modeling. A detailed dual drainage simulation model is described based upon hydraulic flow routing procedures for surface flow and pipe flow. Special consideration is given to the interaction between surface and sewer flow during surcharge conditions in order to most accurately compute water levels above ground as a basis for further assessments of possible damage costs. The model application is presented for a small case study in terms of data needs, model verification and first simulation results. PMID- 16248204 TI - Flooding in the future--predicting climate change, risks and responses in urban areas. AB - Engineering infrastructure is provided at high cost and is expected to have a useful operational life of decades. However, it is clear that the future is uncertain. Traditional approaches to designing and operating urban storm drainage assets have relied on past performance of natural systems and the ability to extrapolate this performance, together with that of the assets across the usable lifetime. Whether or not climate change is going to significantly alter future weather patterns in Europe, it is clear that it is now incumbent on designers and operators of storm drainage systems to prepare for greater uncertainty in the effectiveness of storm drainage systems. A recent U.K. Government study considered the potential effects of climate and socio-economic change in the U.K. in terms of four future scenarios and what the implications are for the performance of existing storm drainage facilities. In this paper the modelling that was undertaken to try to quantify the changes in risk, together with the effectiveness of responses in managing that risk, are described. It shows that flood risks may increase by a factor of almost 30 times and that traditional engineering measures alone are unlikely to be able to provide protection. PMID- 16248205 TI - SIPSON--simulation of interaction between pipe flow and surface overland flow in networks. AB - The new simulation model, named SIPSON, based on the Preissmann finite difference method and the conjugate gradient method, is presented in the paper. This model simulates conditions when the hydraulic capacity of a sewer system is exceeded, pipe flow is pressurized, the water flows out from the piped system to the streets, and the inlets cannot capture all the runoff. In the mathematical model, buried structures and pipelines, together with surface channels, make a horizontally and vertically looped network involving a complex interaction of flows. In this paper, special internal boundary conditions related to equivalent inlets are discussed. Procedures are described for the simulation of manhole cover loss, basement flooding, the representation of street geometry, and the distribution of runoff hydrographs between surface and underground networks. All these procedures are built into the simulation model. Relevant issues are illustrated on a set of examples, focusing on specific parameters and comparison with field measurements of flooding of the Motilal ki Chal catchment (Indore, India). Satisfactory agreement of observed and simulated hydrographs and maximum surface flooding levels is obtained. It is concluded that the presented approach is an improvement compared to the standard "virtual reservoir" approach commonly applied in most of the models. PMID- 16248206 TI - Partnership for sustainable leprosy control beyond 2005. PMID- 16248207 TI - Leprosy in wild armadillos. AB - Wild nine-banded armadillos (Dasypus novemcinctus) in the south central United States are highly endemic natural hosts of Mycobacterium leprae. Surveys conducted over the last 30 years on more than 5000 animals confirm that the infection is present among armadillos in Arkansas, Louisiana, Mississippi and Texas. Highest prevalence rates are found among the animals in low-lying alluvial and coastal areas, primarily in Louisiana and Texas. Both animal density and local factors may contribute to the detectability of armadillo leprosy in those regions. Little evidence for M. leprae infection is found among armadillos elsewhere in the US range, and only a few reports relate finding the infection among animals in Central or South America. However, the issue has received only scant attention in other countries. Armadillos only recently expanded their range into the US, and leprosy was present in Texas and Louisiana prior to the arrival of armadillos. The ecological relationship between humans and armadillos with M. leprae in this region remains unclear. However, infected armadillos constitute a large reservoir of M. leprae and they may be a source of infection for some humans in this country, and perhaps in other locations across the animal's range. PMID- 16248208 TI - Current status of leprosy and leprosy control in Bangladesh: an ongoing collaboration. AB - Elimination of leprosy as a public health problem, defined by a registered prevalence of less than one case per 10,000 population, was achieved by Bangladesh in 1998, and steady reduction in prevalence is ongoing. It is less certain whether a sustained reduction in case detection is occurring, with little overall change in some longstanding programme areas, though the overall annual new case detection rate has fallen by over one-third between 1996 and 2004, from 9.8 to 6.1 per 100,000. Concerns about 'hidden' cases have been raised in Bangladesh as elsewhere, though the National Leprosy Elimination Campaign of 1999 detected relatively fewer new cases than in other countries, and mainly in low endemic areas. Investigation into the correct diagnosis of leprosy and recording and reporting practices has not suggested high levels of over-diagnosis or 'over registration'. Both before and since achievement of the elimination target at national level, the collaboration of Non-Governmental Organizations with the national Leprosy Elimination Program has been considerable. NGOs now support ongoing leprosy control efforts in rural and urban populations, moderate to high endemic for leprosy, amounting to 50% of the entire population, and NGO staff look after 75% of all new cases in Bangladesh. This close collaboration has highlighted the potential for large-scale partnership in disease control, and has expanded to extensive partnership in tuberculosis control, which will hopefully enhance cost-effectiveness and quality of both programmes. Further challenges remain in the area of urban leprosy control, where leprosy case finding represents 30% of the whole country, but public health infrastructure and community organization is weakest. Sustaining of leprosy services in the long term is a significant concern, and new modes of collaboration, with a more technical, supportive role for NGOs in some areas is being piloted. PMID- 16248209 TI - A novel treatment for plantar ulcers in leprosy: local superficial flaps. AB - Plantar ulceration is the most common serious disability occurring in patients of leprosy. Growth and emergence of innovative procedures in plastic surgery has greatly revolutionized the treatment of this highly frustrating problem. In the present study, a total of 40 leprosy patients were included. These ulcers were managed using different types of local superficial flaps (advancement, rotation, transposition and first toe web flap). Majority of ulcers healed within 4 weeks and patients were discharged within 6-8 weeks postoperatively. Patients were followed up for a period of 6 months to 3 years. Ulcers recurred in only 25% of all the ulcers operated upon and we observed that management of plantar ulcers by using appropriate local superficial plantar flaps is a viable option and worth trying in view of low incidence of recurrence and relatively shorter duration of hospital stay. PMID- 16248210 TI - Role of PGL-I antibody detection in the diagnosis of pure neural leprosy. AB - Pure neural leprosy (PNL) is difficult to diagnose because skin lesions and acid fast bacilli (AFB) in slit smears are absent. At present, the gold standard for PNL diagnosis is the histopathological examination of a peripheral nerve biopsy. Even so, detection of bacteria is difficult and histological findings may be non specific. Furthermore, nerve biopsy is an invasive procedure that is only possible in specialized centres. Therefore, there is a need for additional diagnostic methods that may help to confirm the clinical diagnosis of PNL. In the present study, an additional laboratory test, the ELISA for anti-phenolic glycolipid I (PGL-I) IgM antibodies, was performed on 103 individuals with clinical and neurophysiological signs of peripheral neuropathy, of which 67 were diagnosed as PNL patients and 36 remained as 'not diagnosed as PNL', as well as on a control group of 34 patients with other neurological diseases. An antibody response was present in 14/67 (21%) of the patients diagnosed as PNL as compared with 3/34 (9%) of controls. Anti-PGL-I positivity was observed in 5/8 (63%) of the AFB positive cases. Patients whose diagnosis was confirmed solely by Mycobacterium leprae PCR on the nerve sample had 4/25 (16%) seropositivity. In addition, anti-PGL-I antibodies were detected in 9/40 (23%) of the PNL patients who were PCR negative for M. leprae DNA. Moreover, two patients who showed clinical and eletrophysiological manifestations suggestive of PNL were diagnosed with the help of their positive test results in the anti-PGL-I ELISA. In conclusion, detection of antibodies against PGL-I in patients with peripheral neuropathy is useful as an additional laboratory test to help PNL diagnosis. PMID- 16248211 TI - Clinical, histopathological and bacteriological study of 52 referral MB cases relapsing after MDT. AB - Fifty-two BB-LL relapse cases referred to our centre during 1997-2003 were investigated in detail. Twenty-four cases had been treated with extended MB-MDT [until smear negativity (NON-FDT)]. The remaining 28 cases (54%) had received one of the fixed duration regimens (FDT), of whom 11 had 24 months and 6 had 12 months of WHO MB-MDT. Eleven cases had received rifampicin/ofloxacin (RO) treatment. Follow-up slit skin smear reports were available for 41 cases, all but three cases had been smear negative at some point after release from treatment. None of the cases showed any clinical or bacteriological evidence of upgrading, i.e. LL to BT where as downgrading BB to BL occurred in five cases. The duration between cessation of treatment and reappearance of lesions (DCTR) varied from 2 to 15 years. The mean DCTR was longest (9.4 years) for the NON-FDT and 24 months MB-MDT cases. The mean DCTR was significantly lower in the 12 months MB-MDT and RO treated cases (6.8 and 6.2 years, respectively). Four of RO treated cases and four cases with multiple episodes of reaction had DCTR less than 5 years. Inadequate treatment/poor killing of Mycobacterium leprae results in early onset relapse, whereas 'persisting' or 'drug resistant mutants' contribute to late onset relapse. PMID- 16248212 TI - Genital involvement and type I reaction in childhood leprosy. AB - We describe the case of a 4-year-old boy, with a positive family history of multibacillary leprosy (borderline-borderline) in his 12-year-old sister. The patient was diagnosed to have borderline lepromatous (BL) leprosy, BI of 4 + and had two erythematous, infiltrated plaques over the scrotum. He developed type reaction, 3 months following initiation of multibacillary multidrug therapy (MB MDT) and responded favourably to systemic corticosteroids. PMID- 16248213 TI - Reversal reaction and Mitsuda conversion in polar lepromatous leprosy: a case report. AB - A 22-year-old male polar lepromatous leprosy patient who became Mitsuda positive after 36 months of multidrug therapy (MDT) is reported. Lepromatous leprosy (LL) is a state of specific immunosuppression and is invariably irreversible. The finding of Mitsuda positivity in histopathologically proven polar lepromatous leprosy is extremely uncommon, and conversion of lepromin status following MDT has not so far been reported. This case reports confirms the observations made by Waters et al. regarding lepromin conversion in lepromatous patients. PMID- 16248214 TI - Services for leprosy related 'distress': another perspective. PMID- 16248215 TI - Annual variations in Leptospira seroprevalence among sows in southern Vietnam. AB - A serological survey was conducted among sows in the Mekong delta in southern Vietnam in 1999 to investigate variations in leptospiral seroprevalence over a one-year period. In this region, leptospirosis is endemic and a high leptospiral seroprevalence has been shown in the pig population. In this study, the serology of six Leptospira serovars was analysed by the microscopic agglutination test for 429 sows at five large-scale state farms sampled during the dry period, the rainy period and the early dry period. The serovars included were L. interrogans serovar (sv) autumnalis strain Akiyama A, L. interrogans sv bratislava strain Jez, L. interrogans sv icterohaemorrhagiae strain Kantorowicz, L. interrogans sv pomona strain Pomona, L. borgpetersenii sv tarassovi strain Perepelitsin, and L. kirschneri sv grippotyphosa strain Duyster. Variations in seroprevalence over the year were found for sv bratislava and sv icterohaemorrhagiae: the seroprevalence was higher during the dry period compared with the rainy period (p = 0.07 and p = 0.005, respectively) and the early dry period (p = 0.00006 and p = 0.0006, respectively). It is concluded that in regions where water is constantly abundant and where animals are exposed to the outdoor environment all year round there are highly significant variations in leptospiral seroprevalence over the year. PMID- 16248216 TI - Clinicopathological indications of resilience of swamp buffaloes to infection with Fasciola gigantica. PMID- 16248217 TI - Evaluation of diagnostic tests for Trypanosoma evansi in experimentally infected pigs and subsequent use in field surveys in north Vietnam and Thailand. AB - This study is concerned with the evaluation of established diagnostic tests for diagnosis of Trypanosoma evansi in pigs. The immune trypanolysis test (TL), card agglutination test (CATT), latex agglutination test (LATEX), enzyme-linked immunosorbent assay (ELISA), microhaematocrit centrifugation technique (MHCT) and mouse inoculation (MI) tests were initially evaluated in experimentally infected fattening pigs. All infected pigs were confirmed parasitologically positive with both MHCT and MI. Results of the serological assays indicated that the TL could be a reference test for the presence of RoTat 1.2 antibodies in pigs. The results of the CATT and LATEX were inconsistent with the TL while the ELISA results correlated with the TL results. The four serological assays were subsequently used in two field surveys in Vietnam and Thailand. Results of the two agglutination assays (CATT and LATEX) were not consistent and did not correlate with TL results. The ELISA at percentage positivity of 22 appeared to have good ability to discriminate between seropositive and seronegative animals. Of the 437 samples collected at smallholder pig premises in northern Vietnam, no positive pigs were detected with the TL test. In Thailand, 77 samples were collected from five farrowing farms with a history of surra. Two parasitologically positive sows were found and on each farm seropositive sows were detected. PMID- 16248218 TI - Effects of a novel pesticide resistance management strategy on tick control in a smallholding exotic-breed dairy herd in Kenya. AB - Effects of a novel pesticide resistance management strategy on tick control are evaluated in this study. The study is based on a temporal analysis of tick management practices on a smallholding in western Kenya. Results are reported of an innovation to tackle individual resistance in a pair of alternative pesticides using relay application. Incidence of tick-borne diseases at the farm were reduced from 79.6% per annum to 4.5% and no cases were observed in the last two years of the study. Negative cross-resistance is believed to be the mechanism in play for this effective tick control practice. Tick-borne disease control and management costs were halved in comparison to application of a single ineffective pesticide at the same treatment frequency. The acaricide relay strategy is suitable for smallholdings and is expected to significantly extend the useful lifespan of the pesticide pair. PMID- 16248219 TI - Strategies, effectiveness and rationale of vector-borne disease control in the pastoralist system of south-western Uganda. AB - In Uganda, control of vector-borne diseases is mainly in form of vector control, and chemotherapy. There have been reports that acaricides are being misused in the pastoralist systems in Uganda. This is because of the belief by scientists that intensive application of acaricide is uneconomical and unsustainable particularly in the indigenous cattle. The objective of this study was to investigate the strategies, rationale and effectiveness of vector-borne disease control by pastoralists. To systematically carry out these investigations, a combination of qualitative and quantitative research methods was used, in both the collection and the analysis of data. Cattle keepers were found to control tick-borne diseases (TBDs) mainly through spraying, in contrast with the control of trypanosomosis for which the main method of control was by chemotherapy. The majority of herders applied acaricides weekly and used an acaricide of lower strength than recommended by the manufacturers. They used very little acaricide wash, and spraying was preferred to dipping. Furthermore, pastoralists either treated sick animals themselves or did nothing at all, rather than using veterinary personnel. Oxytetracycline (OTC) was the drug commonly used in the treatment of TBDs. Nevertheless, although pastoralists may not have been following recommended practices in their control of ticks and tick-borne diseases, they were neither wasteful nor uneconomical and their methods appeared to be effective. Trypanosomosis was not a problem either in Sembabule or Mbarara district. Those who used trypanocides were found to use more drugs than were necessary. PMID- 16248221 TI - Effect of replacement of maize with cassava peel in cockerel diets on performance and carcass characteristics. PMID- 16248220 TI - Theileriosis in sheep and goats in the Sultanate of Oman. PMID- 16248222 TI - Reduced milk production in udder quarters with subclinical mastitis and associated economic losses in crossbred dairy cows in Ethiopia. AB - The objective of the study was to estimate the losses associated with subclinical mastitis (SCM) in crossbred dairy cows in the Central Highlands of Ethiopia. A split udder investigation was performed with 30 cows to determine production losses associated with SCM. Each quarter of the study cows was examined using the California Mastitis Test (CMT) and quarter milk production was measured over a period of 8 days. Production losses were determined for different CMT scores by comparing production of quarters with CMT score 0 to quarters with CMT scores trace, 1, 2 and 3, respectively. Using data from a recently published study, economic losses were determined for different farm sizes and production subsystems by multiplying the prevalence of the respective CMT scores with the production losses associated with these CMT scores. Mean quarter milk production was 0.82 +/- 0.40 kg per milking in the split udder trial. Milk production was reduced by 1.2%, 6.3%, and 33% in quarters with CMT scores 1+, 2+, and 3+, respectively. Using data from the published study, a quarter with SCM lost an average of 17.2% of its milk production. Production losses associated with SCM were estimated at 5.6% for the Addis Ababa Milk Shed. Stratified losses were highest (9.3%) in urban dairy farms (UDF) and small-scale farms (6.3%). The estimates of the financial losses ranged from US dollars 29.1 in dairy herds in secondary towns (DHIST) to US dollars 66.6 in UDF. A total loss of US dollars 38 was estimated for each cow per lactation. Reducing mastitis in UDF (highest prevalence) to the level of DHIST (lowest prevalence) could reduce the loss by US dollars 35. As this does not include costs associated with treatment or culling of diseased cows, this figure probably underestimates the possible benefits of control measures. PMID- 16248224 TI - Biochemical and haematological measurements in beef cattle in Mendoza plain rangelands (Argentina). AB - Biochemical and haematological measurements were used to identify constraints on productivity in beef cattle. One hundred and twelve Aberdeen Angus and Criollo Argentino females including lactating cows, dry non-pregnant cows and heifers were selected. Blood samples were taken in the middle of summer and autumn. Serum was analysed for haemoglobin, PCV, glucose, albumin, urea, creatinine, Ca, P, Mg, Fe, Cu, ALP, AST, ALT, CK, LDH, Cl-, Na and K content. Globulin was calculated by taking the difference between total protein and albumin. Percentages of all cattle that had metabolite levels outside reference ranges were: 15% (glucose), 8% (globulin), 5% (urea), 96% (Ca), 50% (P), 12% (Mg), 20% (Na), 5% (K), 24% (Cl), 18% (Fe), 5% (Cu) and 85% (CK). These results indicate with some certainty that dietary protein was not limiting. Body condition score loss was detected only in lactating cows. There were a few animals that could have presented chronic inflammatory disease. Phosphorus could be an important potential constraint on fertility, although the presence of symptoms of hypophosphataemia was not observed. The study also demonstrates the absence of anaemia or liver disease. Breed, seasonal and physiological state differences in some blood metabolites could be attributed to one or more of the following factors: chemical composition of the feed ingested, environmental temperature, nutrient content of the forage, animal age and cattle foraging experience. The study provides a basis for implementing helpful adjustments in current cattle management practices so as to alleviate the constraints on productivity, provided that these practices are profitable. PMID- 16248223 TI - A cross-sectional study of reproductive performance of smallholder dairy cows in coastal Tanzania. AB - A retrospective cross-sectional study was conducted on 200 randomly selected smallholder farms from a mixed dairy farming system in Tanga, Tanzania, between January and April 1999. We estimated the frequency and determinants of long calving interval (LCI), retention of fetal membrane (RFM), dystocia, and abortion in smallholder crossbred cattle and explored birth trends. The mean calving interval was 500 days and birth rate was 65 per 100 cow-years. Dystocia was reported to affect 58% of calvings, and 17.2% of animals suffered RFM. Using mixed effect models, the variables associated with LCI, RFM and dystocia were breed, level of exotic blood and condition score. Zebu breeding was associated with LCI (odds ratio (OR) = 2.3, p = 0.041) and Friesian breeding with lower odds for RF (OR = 0.26, p = 0.020). Animals with higher levels of exotic blood had lower odds for evidence of dystocia (OR = 0.45, p = 0.021). Evidence of dystocia was significantly associated with poor condition score (p = -1.10, p = 0.001). Our observations suggest that LCIs are common in smallholder dairy farms in this region and a likely source of economic loss. Dystocia, RFM, poor condition score and mineral deficiency were common problems and were possibly linked to LCI. PMID- 16248225 TI - Health risk management: well-being for the employee and the bottom line. AB - For most employers, a small percent of the employee/participant population accounts for a large percent of health care costs. However, the population of this high-cost group changes from year to year. The fundamental problem is keeping employees out of the high-risk/high-cost segment, something plan design changes cannot address but that integrated health risk management (IHRM) can help achieve. This article explains how employers can implement an IHRM program to significantly lower health care costs to a degree unattainable through traditional cost-control strategies, while simultaneously raising workers' productivity and well-being. PMID- 16248226 TI - Population health improvement--the next era of the health care management evolution. AB - The trend of increasing health care costs over the past several years shows no signs of slowing down. While employers have attempted to address the issue with various cost-shifting and cost-sharing initiatives to relieve the burden, those attempts have often fallen short. In today's fiscally scrutinized benefits environment, the emphasis needs to be more strategic. The management of employee health and productivity needs to move to a more encompassing organizational view that addresses the most expensive drivers of these costs head-on, with prevention, education and employee responsibility as key tenets. Organizations that make the move--by implementing programs to maintain, improve and manage their population's health--will enjoy substantial cost savings and enhanced employee productivity. PMID- 16248227 TI - A pivotal year for managing retiree medical benefit costs. AB - With Medicare set to begin offering a prescription drug benefit in 2006, employers that provide medical coverage to retirees aged 65 and older have a unique opportunity to redefine their programs and financial commitments for the long term. The new Medicare Part D drug benefit poses a range of alternatives for employers to consider--from eliminating post-65 drug benefits altogether to maintaining or modifying their current programs to qualify for the 28% federal tax subsidy for eligible drug costs. In deciding on a course of action, companies need to consider a host of complex issues, including workforce needs and demographics, employee relations, plan design and administration--in addition to the long-term financial and accounting implications. This article reviews the alternatives available to employers with the advent of Medicare Part D and highlights some of the key factors employers should consider as part of this decision. PMID- 16248228 TI - Are employers pursuing value-based purchasing? AB - Some employers have begun to undertake a variety of initiatives, collectively termed value-based purchasing (VBP), which aim at factoring quality into the purchasing decision-making process when negotiating costs with providers and insurers. There is evidence that the VBP movement has begun gaining momentum; yet it is unclear whether employers are firmly committed to VBP principles. By surveying a national sample of employers, this study investigates the extent to which employers are currently engaged in VBP activities and also increases our understanding of barriers hampering the broader implementation of these activities. PMID- 16248230 TI - Subrogation/reimbursement--health plans. Westaff (USA) Inc. v. Arce. PMID- 16248229 TI - The American Jobs Creation Act and its impact on deferred compensation: reassessment from a business perspective. AB - The American Jobs Creation Act (AJCA), which was signed into law in October 2004, will have an impact on almost every deferred compensation program in the United States. This article argues that as companies continue to evaluate the transition alternatives under AJCA and contemplate the necessary changes to the plan program, companies also should consider simultaneously addressing broader issues surrounding nonqualified deferred compensation arrangements. These include ongoing business purpose, financial planning considerations, education of participants, corporate governance considerations and the potential implications to international assignees. PMID- 16248232 TI - Federal jurisdiction--complete preemption vs. conflict (ordinary) preemption. Arana v. Ochsner Health Plan, Inc. PMID- 16248231 TI - Federal jurisdiction--preemption--state antisubrogation law--state law regulating insurance saved from preemption. Singh v. Prudential Health Care Plan. PMID- 16248233 TI - Preemption--common law fraud. Trustees of the AFTRA Health Fund v. Biondi. PMID- 16248234 TI - [Study on objectivation of syndrome typing of jaundice]. AB - OBJECTIVE: To investigate the essence of dampness and heat, the two main etiological factors of jaundice, to seek the objectified indices for syndrome typing of jaundice. METHODS: Ninety-five adult in-patients with icteric viral hepatitis B were eligible, 61 of them were free from bacterial or fungal infection, while 34 with secondary bacterial or fungal infection. The serum biochemical indexes, and clinical features such as the color of jaundice, tongue picture and pulse frequency were observed. RESULTS: Dark-yellow colored skin, red or normal tongue proper with white tongue fur appeared in most patients with dominant elevation of direct bilirubin, in whom the ratio of direct bilirubin to indirect bilirubin (DBIL/IBIL > or =1). The color of tongue proper often turned pale in patients with cirrhosis due to the complicated anemia caused by hyperactivity of spleen, while the quick pulse and yellow tongue fur always presented in patients accompanied with infection. On the contrary, bright-yellow colored skin, red tongue with yellow fur appeared in patients with dominant elevation of indirect bilirubin, in whom DBIL/IBIL <1. CONCLUSION: DBIL/IBIL ratio can exactly reflect the syndrome types of jaundice, when it is equal to or above 1, the type is mostly of dampness dominant, whereas it is less than 1, heat dominant. If quick pulse and yellow tongue fur appeared in patients with DBIL/IBIL > or =1, it indicates that the presence of heat conversion of the dampness stagnancy. PMID- 16248236 TI - [Intervention of maiwei dihuang oral liquid on hormonotherapy in treating active systemic lupus erythematosus]. AB - OBJECTIVE: To observe the intervention of Maiwei Dihuang Oral Liquid (MDOL) on hormonotherapy in treating active systemic lupus erythematosus (SLE). METHODS: Sixty SLE patients in active stage were randomly and equally allocated into two groups, and administered with prednisone, which was medicated in initial dose of 0.5-1 mg/kg, and with the dose being reduced conditionally 6-8 weeks. To the 30 patients in the treated group 10 ml MDOL twice daily was given additionally. The therapeutic course was 3 months. RESULTS: The therapeutic effect in the treated group was better than that in the control group (P < 0.05). Systemic lupus erythematosus disease activity index (SLEDAI) was significantly improved in both groups (P < 0.01), but was superior in the treated group (P < 0.05). The dose of prednisone used was significantly reduced (P < 0.01), and the scores of Yin deficiency fire-flourishing syndrome were obviously decreased (P < 0.01) in the treated group while in the control group, these indexes were unchanged (P > 0.05), the difference between the two groups was significant (P < 0.01). The occurrence of adverse reaction was significantly lower in the treated group than that in the control group (P < 0.05). CONCLUSION: MDOL can obviously improve the effect of hormonotherapy in SLE, it has advantages in reducing the dosage used and antagonizing the adverse reactions of glucocorticoid. PMID- 16248235 TI - [Relationship between basal core promoter combined point mutation of hepatitis B virus and TCM syndrome type]. AB - OBJECTIVE: To investigate the relationship between basal core promoter (BCP) combined point mutation of hepatitis B virus (HBV) and TCM syndrome type. METHODS: One hundred and two patients with chronic hepatitis with positive HBV DNA and hadn't ever been treated by Lamivudine and interferon were differentiated according TCM syndrome differentiation into 5 types, two excess types (damp-heat blocking zhong-jiao type and blood stasis blocking collaterals type) and three deficiency types, gan-stagnation with pi-dificiency type, gan-shen yin-deficiency type and pi-shen yang-deficiency type. The serum HBV DNA, hepatic biochemical indexes, and the mutation of BCPnt 1762A-T and nt1764G-A combined point were determined, respectively. RESULTS: The variant strain positive rate detected in the excess type was significantly higher than that in the deficiency type, the highest rate appeared in patients of damp-heat blocking zhong-jiao type. CONCLUSION: BCP combined point mutation may be liable to happen in patients of TCM excess type, especially in patients of damp-heat blocking zhong-jiao type. PMID- 16248237 TI - [Effect of shensong yangxin capsule on ventricular premature beat and cardiovascular autonomic nervous function in patients with coronary heart disease]. AB - OBJECTIVE: To observe the clinical efficacy of shensong yangxin capsule (SYC) on ventricular premature beat (VPB) differentiated in TCM as palpitation of Qi-yin deficiency syndrome or Xin collateral stagnation syndrome, and cardiovascular autonomic nervous function in patients with coronary heart disease (CHD). METHODS: The randomized, double-blind, parallel contrast method was adopted, patients were randomly assigned by 3:1 ratio into two groups. One hundred and sixty-five patients in SYC treated group and 56 in the control group (treated with Xinlvning tablet), and the therapeutic course for both groups was 4 weeks. RESULTS: The clinical efficacy on VPB and in improving TCM syndromes was better in SYC group than that in the control group (P < 0.01). After treatment, the heart rate variability (HRV) and QT dispersion in the two groups were improved in a certain degree. The changes of SDNN, SDANN, SDNN Index and PNN50 in the two groups were significantly different (P < 0.05, P < 0.01), the efficacy in the treated group was superior to that in the control group. CONCLUSION: SYC has definite effect on VPB and TCM Syndromes, it can obviously meliorate the activity of cardiovascular autonomic nervous system in the patients with CHD. PMID- 16248238 TI - [Observation on effect of compound danshen droplet-pill combined with trimetazidine in treating senile unstable angina pectoris]. AB - OBJECTIVE: To investigate the effect of compound Danshen Droplet-pill (DS) combined with trimetazidine (TMZ) in treating senile unstable angina pectoris (SUAP). METHODS: One hundred and twenty patients with SUAP were eaually and randomly divided into 2 groups, the treated group and the control group. Changes of angina, occurrence of arrhythmia, myocardial infarction and sudden death, myocardial ischemia in ECG and partial indexes of heart function were observed. RESULTS: The total effective rate in the treated group was 78.3%, while that in the control group was 53.3%, comparison of the two groups showed significant difference (P < 0.05). The incidence rate of arrhythmia in the two groups was 18.2% and 30.0% respectively and that of acute myocardial infarction and sudden death was 0 and 5.0% respectively, also showed significant difference between them (P < 0.05). Condition of myocardial ischemia revealed in ECG and partial indexes of heart function in the treated group were all improved to certain extent. Conclusion DS combined with TMZ is superior in treating SUAP. PMID- 16248239 TI - [Clinical observation on effect of modified huanglian wendan decoction in treating diabetic asymptomatic myocardial ischemia]. AB - OBJECTIVE: To investigate the clinical efficacy of modified huanglian wendan decoction (MHWD) on diabetic asymptomatic myocardial ischemia. METHODS: Ninety patients were randomly divided into two groups. The control group (n=30) was given Xinkang tablet (XKT) at a dose of 20 mg, twice a day. The treated group (n=35) was given MHWD besides XKT as that given to the control group. The treatment course for both groups was 1 month. Indexes, including blood glucose, glycosylated hemoglobin, blood lipids, myocardial zymogram, hemorheologic parameters, urinary albumin, routine examination of blood and urine, function of liver and kidney, as well as 24h dynamic electrocardiogram and electrocardiogram exercise test were measured in the two groups before and after treatment. RESULTS: The total clinical effective rate in the treated group and the control group was 88.33% and 56.67% respectively (P < 0.05), showing significant difference between them. The frequency of ischemia attacking, paroxysmal cumulative time, motion related incidence were lower in the treated group after treatment than those in the control group. Besides, in the treated group after treatment, the level of blood lipids and hemorheologic parameters were significantly improved (P < 0.05 or P < 0.01), hematocrit was unchanged and triglyceride, red blood cell agglutination index and erythrocyte deformability index were obviously different to those in the control group (P < 0.05). While in the control group after treatment, except the improving of whole blood viscosity (P < 0.05), no significant change was found in the other indices. CONCLUSION: MHWD has effects in improving myocardial ischemia, bettering hemorheologic condition and reducing blood lipids. PMID- 16248240 TI - [Clinical research on combined therapy of diane-35 with modified yougui pill on polycystic ovarian syndrome]. AB - OBJECTIVE: To investigate the clinical efficacy of integrative traditional Chinese and Western medicine on polycystic ovarian syndrome (PCOC). METHODS: Sixty-three patients with polycystic ovarian syndrome were randomly allocated into 2 groups, 31 patients orally administered with diane-35 in the Western medicine group (WMG) and 32 patients treated with conventional controlled medicine plus modified Yougui Pill in the integrative medicine group (IMG). Changes of relevant hormones and clinical syndromes in patients were detected before treatment, after 3 cyclic treatment and at the 6th cycle after treatment. RESULTS: The levels of relevant hormones and the indexes of B-ultrasonic were obviously improved after treatment in the two groups. But 6 cycles after treatment, these changes restored to the baseline as those before treatment in WMG, while maintained in IMG (P < 0.01). What's more, the normalization of menstruation, ovulation and pregnancy rate in IMG were significantly higher than those in WMG (P < 0.01). CONCLUSION: Diane-35 combined with modified yougui pill in treating polycystic ovarian syndrome not only shows marked short-term effect, but could consolidate the curative effect. PMID- 16248241 TI - [Clinical observation on treatment of 18 patients with pulmonary infection after renal transplantation by integrative traditional Chinese and Western medicine]. AB - OBJECTIVE: To observe the therapeutic effect of integrative traditional Chinese and Western medicine in treating patients with pulmonary infection after renal transplantation. METHODS: Thirty-four inpatients were randomly divided into the treated group (n=18) and the control group (n=16). They were treated with conventional therapy, including corticosteroid, anti-viral, anti-bacterial, symptomatic and supporting therapy, to the treated group, the modified qingwen baidu decoction (MQBD) was administered additionally. RESULTS: Fifty of the 18 patients in the treated group were cured, 2 improved and 1 died, the cure rate being 83.3% and the total effective rate 94.4%; while 8 of the 16 patients in the control group were cured, 2 improved and 6 died, the cure rate being 50.0% and the total effective rate 62.5%, comparison between these two groups showed significant difference (P < 0.05). CONCLUSION: Promising effect could be obtained for treatment of patients with pulmonary infection after renal transplantation by adding MQBD, a Chinese herbal medicine for clearing heat and detoxication, cooling blood and removing fire, on the basis of conventional western medical treatment. PMID- 16248242 TI - [Clinical study on treatment of obsessive compulsive neurosis by acupoint stimulating control]. AB - OBJECTIVE: To study the clinical effect of acupoint stimulating control (ASC) in treating obsessive compulsive neurosis. METHODS: The comparative study was conducted in 65 patients with obsessive compulsive neurosis, they were divided into two groups, the 33 patients in the control group treated with chlorimipramine and the 32 in the tested group treated with ASC. The therapeutic efficacy and adverse reaction were assessed according to the standard for clinical efficacy evaluation by Yale-Brown scale for obsession (Y-BOCS)and adverse reaction scale. RESULTS: The curative rate and markedly effective rate in the control group was 24.2% (8/33) and 27.3% (19/33), which in the tested group was 37.5% (12/32) and 34.4% (11/32) respectively. Significant difference was shown in comparison of Y-BOCS score between the two groups from the end of the 4th week of treatment (P < 0.05), indicating the efficacy in the tested group was better than that in the control group. Moreover, the occurrence of adverse reaction was higher in the control group than that in the tested group. CONCLUSION: ASC is a treatment with good effect, less adverse reaction and favourable safety superior to the treatment by chlorimipramine. PMID- 16248243 TI - [Experimental study on effect of baoganning on activity of nuclear transcription factor-kappaB in hepatic stellate cells]. AB - OBJECTIVE: To investigate the effect of Baoganning (BGN) on activity of nuclear transcription factor-kappaB (NF-kappaB) in hepatic stellate cells (HSCs) and its relevant mechanisms. METHODS: Normal Wistar rats were medicated with BGN decoction by gavage for 7 days to prepare BGN drug-serum. The effect of BGN drug serum on HSC-T6 growth was measured by MTT assay; phosphorylation level of NF kappaB inhibiting factor IkappaB at different time after BGN stimulation was detected by Western blotting analysis; and the binding level of NF-kappaB with DNA was measured 30 min after drug-serum stimulation with gel shift assay. RESULTS: BGN could significantly inhibit the HSC-T6 growth and quickly supress the phosphorylation of IkappaB, with the effect reached its peak at 30 min and restored to baseline level 6 h after stimulation, and reduce the binding capacity of NF-kappaB with DNA. CONCLUSION: BGN can inhibit phosphorylation of IkappaB, restrain the activity of NF-kappaB and change the binding level of NF-kappaB with DNA. PMID- 16248245 TI - [Comparative study on changes of digestive and absorptive functions in three different models of Pi-deficiency syndrome]. AB - OBJECTIVE: To compare the changes of digestive and absorptive functions in three different models of Pi-deficiency syndrome (PDS). METHODS: Experimental mice were divided into four groups, the control group(CG), the rhubarb group (RG), the exhaustion group (EG) and the over-exertion group(OG). Criteria including general physical signs, D-xylose excretion rate, serum amylase activity, velocity of stomach emptying and enterokinesia, serum gastrin content and indexes of organs were determined before and after treatment. RESULTS: Decrease of D-xylose excretion rate and increase of stomach emptying and enterokinesia velocity appeared in all the three PDS models. As compared with CG, changes of all indices in OG were significant, while the decreasing of spleen index and serum amylase activity in EG, and the changes of serum gastrin content and thymus index in RG were insignificantly different. CONCLUSION: All the changes in various criteria showed that PDS mice model established by over-exertion was superior to that established by frequently used methods as purging with rhubarb and exhausting by swimming. PMID- 16248244 TI - [Differential expression and influence of caoguo zhimu decoction on kindling epilepsy related genes]. AB - OBJECTIVE: To screen the relative genes associated with the genesis of kindling epilepsy and the effect of caoguo zhimu decoction (CZD) on these genes. METHODS: mRNA differential display reverse transcription PCR was used to screen differential gene bind, and Northern blotting hybridization was applied to exclude the false positive reaction. RESULTS: There were as more as 110 differential expression bindings in the normal group, model group and the treated group, among them 11 binds with differential expression only appeared in the model group but not in the normal group and disappeared in the treated group. It indicated that the 11 genes are correlated with the genesis and treatment of kindling epilepsy. By searching in the Gene-bank of NCBI, 7 bindings of the 11 were homologous genes and 4 gene fragments are novel genes with unknown function, which have been registered at the Gene-bank, with the registered numbers of CK325391, CK325392, CK325393, CK325394, respectively, and false positive possibility of 3 novel genes was excluded. CONCLUSION: CZD has effect in treating kindling epilepsy may be through influencing the expression of partial gene fragments in the hippocampus. PMID- 16248246 TI - [Preventive effect of multi-glycoside of tripterygium Wilfordii Hook. f. on proteinuria and mesangial injury in experimental mesangial proliferative glomerulonephritis]. AB - OBJECTIVE: To observe the preventive effect of multi-glycoside of Tripterygium Wilfordii Hook. f. (GYW) on proteinuria and mesentery injury in experimental mesangial proliferative glomerulonephritis in vivo. METHODS: The reversible anti Thyl.1 antibody glomerulo nephritis model of rats was established with monoclonal antibody 1-22-3 and intervened with GTW, and a control group was set up in the same time. Changes of 24h urinary protein excretion, serum creatinine (Scr), blood urea nitrogen (BUN), total plasma protein (TP) and glomerular morphology were observed, and the level of mRNA expression of proliferative factors, including platelet-derived growth factor-BB (PDGF-BB) and transforming growth factor-beta (TGF-beta), in renal tissue was determined. RESULTS: GTW could inhibit proteinuria and mesangial injury in anti-Thyl. 1 antibody nephritis model. The PDGF-BB and TGF-beta mRNA expression in the anti-Thy1.1 antibody nephritis model rats were increased for 2.84 and 1.64 times respectively to those in the normal control group. GTW could down-regulate the over-expression of PDGF BB mRNA by 33.1%, it was significantly different to that in the control group (P < 0.05). CONCLUSION: GTW could reduce the proteinuria and inhibit mesangial cells proliferation and extracellular matrix deposition, these effects maybe related to the down-regulating of PDGF-BB mRNA expression. PMID- 16248247 TI - [Study on enhancing bioavailability of paeoniflorin by combined use with Chinese herbs for inner-warming]. AB - OBJECTIVE: To study the influence of Chinese herbs for inner-warming on the bioavailability of paeoniflorin (PF) and its mechanism. METHODS: Chinese herbs (pepper fruit, evodia fruit, cassia bark, fennel fruit and prickly-ash peel) were separately used in combination with PF for gastrogavage to mice. Reversed phase high-performance liquid chromatography was used to determine the plasma concentration of PF in mice after medication. The bioavailability of PF was calculated and compared, taking single use of red peony root for control. RESULTS: The pharmacokinetics of PF in mice was conformed to the one-compartment model, as combined use with Chinese herbs for inner-warming, the relative bioavailability of PF was 137.22% for pepper fruit, 123.62% for evodia fruit, 108.39% for cassia bark, 226.02% for fennel fruit and 116.73% for prickly-ash peel, there were difference of Cmax and AUC(0-infinity) in comparison of these data with the control group (P < 0.05), but with no difference of tmax (P > 0.05). CONCLUSION: The Chinese herbs used in this experiment in combination with red peony root could enhance the bioavailability of PF, which illustrated the scientific meaning of the recipe combination of Chinese herbs for activating blood circulation and inner-warming viewing from pharmacodynamics. PMID- 16248248 TI - [Study on mechanism of the anti-tumor activity of Acanthopanax gracilistylus]. AB - OBJECTIVE: To study the mechanism of anti-tumor activity of Acanthopanax gracilistylus extract (Age). METHODS: The tumor cells proliferation was detected by using (3H)-TdR incorporation method, and the effects of Age on cell cycle of tumor cells, retinoblastoma (Rb) protein and cyclin-dependent kinases (Cdk) were analyzed by flow cytometry and Western blotting assay, respectively. RESULTS: It was indicated by cytoactivity test in vitro that Age only had effect in inhibiting the proliferation of tumor cells, it couldn't lead to death of cells. Under action of Age, the proliferation of tumor cells was halted at G0/G1 stage of cell cycle, and showed no direct cytotoxic effect by Age. Age could induce lowering of the expression of Rb, Cdk2 and Cdk4, cause halt of tumor cell proliferation. CONCLUSION: The tumor inhibitory effect of Age is realized by way of regulating the activity of cell cycle controlling enzymes to suspend the proliferation of tumor cells. PMID- 16248249 TI - [Effect of TCM therapy by principle for nourishing Qi, activating blood circulation and resolving phlegm on quality of life of patients with chronic obstructive pulmonary disease]. AB - OBJECTIVE: To observe the effect of TCM therapy, by principle for nourishing Qi, activating blood circulation and resolving phlegm, on quality of life of patients with chronic obstructive pulmonary disease (COPD). METHODS: Seventy-two COPD patients with Qi-deficiency, blood stasis and phlegm retention syndrome were randomly divided into two groups. Thirty-five patients in the treated group were treated with the integrative traditional and Western medicine, that is, conventional Western medicine combined with Chinese compound recipe prescribed based on the principle of nourishing Qi, activating blood circulation and resolving phlegm, or with Feikang granule and Bailing capsule, while 37 in the control group was treated with conventional Western medicine alone. The clinical efficacy was evaluated by assessment software of COPD quality of life after patients were treated for 3 months. RESULTS: The clinical efficacy in the treated group was superior to that in the control group, showing significant difference (P < 0.01). CONCLUSION: Compound recipe prescribed according to the therapeutic principle for nourishing Qi, activating blood circulation and resolving phlegm can improve the symptoms of patients with COPD at stable stage, elevate their quality of life. PMID- 16248250 TI - [Clinical observation on effect of electro-acupuncture on back-shu points in treating chronic fatigue syndrome]. AB - OBJECTIVE: To explore the clinical efficacy of electro-acupuncture (EA) on 5 visceral Back-shu points in treating chronic fatigue syndrome (CFS). METHODS: Fourty patients with CFS were treated by EA on Back-shu points, the changes of scoring by Fatigue Assessment Instrument (FAI) and Symptom Checklist 90 (SCL-90) were observed before and after treatment. RESULTS: The scale of FAI and SCL-90 significantly decreased after EA (P < 0.01). CONCLUSION: EA on Back-shu points is one of the effective approaches for treatment of CFS. PMID- 16248251 TI - [Clinical study on effects of kangwei granule on precancerous lesion in patients with chronic atrophic gastritis]. AB - (GPL) in patients suffered from chronic atrophic gastritis (CAG) differentiated as Pi-deficiency with damp-heat retention and blood stasis in TCM Syndrome differentiation. METHODS: Sixty-eight patients fitting to the admission criteria were randomly divided into two groups, 36 patients were treated with KWG in the treated group and 32 were treated with Weifuchun in the control group, all were treated for 2 treatment courses (12 weeks as one course). RESULTS: The curative effects on gastroscopy and pathologic changes in the treated group were significantly superior to those in the control group (P < 0.05). The comparison of clinical efficacy, symptom improvement, anti-Helicobactor pylori effect between the two groups was insignificantly different (P > 0.05). CONCLUSION: KWG is an effective drug for GPL. PMID- 16248253 TI - [Pondering on syndrome differentiation of diseases]. AB - The disease classification in Western medicine and syndrome differentiation in traditional Chinese medicine were comparatively analyzed in this paper. It is proposed that the research on syndrome differentiation of disease classification is one of the cut-in points in integrative medicine research, and indicated that the research may be deepened by way of TCM syndrome classification based on diagnosis and on therapeutic efficacy. PMID- 16248252 TI - [Treatment of ulcerative colitis by combined therapy of retention enema and per colonoscopic spraying with zhikang capsule compound liquid]. AB - OBJECTIVE: To explore the therapeutic efficacy of patients with ulcerative colitis (UC) treated by retention enema and per-colonoscopic spraying of Zhikang Compound Liquid (ZKCL). METHODS: Eighty-six patients with UC were divided into two groups. The 52 patients in the treated group were treated for 4 courses of retention enema, the drug for enema used in the 1st course was ZKCL-A (consisted of normal saline, Zhikang capsule, gentamycin and dexamethasone) and smecta, in the 2nd course ZKCL-A alone, in the 3rd and 4th course, ZKCL-B (with the same contents of ZKCL-A but without dexamethasone), the enema was carried out once a day in the evening, 15 days as one course. Besides, local spraying of ZKCL-A and smecta were given once by colonoscopy before the 1st and 3rd course. The 34 patients in the control group were treated by salicylazosulfapyridine orally. RESULTS: In the treated group, 32 patients got complete remitted, 15 were treated effectively, 5 ineffectively, the total effective rate being 90.38% while the corresponding number in the control group were 8, 14, 12, and 64.71%, respectively. Significant difference was seen when compared with the therapeutic effects of the two groups. CONCLUSION Good efficacy was got in treating patients with UC by retention enema and per-colonoscopic spraying with ZKCL. PMID- 16248254 TI - [Occurrence and prevention of adverse reaction induced by puerarin injection]. AB - In this paper, literature about adverse reaction induced by Puerarin injection published in recent years were reviewed and analyzed. It was reported that Puerarin injection can not only induce adverse reactions on multiple systems, such as immune, blood, urinary, digestive and cardiovascular system, but so far as to cause death. Therefore, it should be administered rationally and strictly according to the direction and the course of medication should be rigorously monitored to prevent the occurrence of adverse reactions. PMID- 16248255 TI - [On establishment and assessment of experimental vulnerable plaques animal models]. AB - Vulnerable plaques is the hot spot in the researching field of cardiovascular diseases. In this paper, literature about establishment of experimental vulnerable plaques model animals published recent years were briefly reviewed and introduced concretely the conception, significance of researching, histopathologic characteristics of various types model, model assessment and current status of research. PMID- 16248256 TI - [Survey of study on therapeutic mechanism of chronic obstructive pulmonary disease treated by traditional Chinese medicine]. AB - This paper reviewed the reports, published in recent years in China, concerning the mechanisms of Chinese herbal medicines in treating chronic obstructive pulmonary disease (COPD), trying to find out its preponderant targets and links, for elevating the level of TCM for prevention and treatment of COPD. PMID- 16248257 TI - [Diversity, inequality and health: the challenge in the face of complexity]. PMID- 16248258 TI - [Determinants of hypercholesterolemia in Geneva: should the focus be on genes or environment?]. AB - Do genetic (G), or environmental (E), factors primarily determine blood lipid concentrations in the Geneva population? On-going research on the causes of hypercholesterolemia in populations based on data collected by the Bus Sante Survey is summarized. About 2/3 of the HDL-cholesterol variance could not be explained by the most important 5 of 10 environmental factors and 9 of 275 common genetic variants identified in the analyses. The remaining 1/3 of the variance was explained mainly by obesity, smoking, alcohol intake, age, and gender. The common polymorphisms played a much smaller role. GxG, GxE, and ExE interactions were the weakest determinants. Environmental factors appear to be the main determinants of hypercholesterolemia in populations. Measurement of genetic traits for clinical or public health purposes is currently not useful. PMID- 16248259 TI - [Interdisciplinary, an undisciplined practice?]. AB - This article aims at better grasping the stakes of interdisciplinary work. The interdisciplinary and integrative clinical approach used at the "Consultation interdisciplinaire de medecine et de prevention de la violence") (CIMPV) to address issues of violence is based on the theories of complexity and built upon the confrontation of personal, professional, intra- and inter-institutional logics. It offers a process that is original in terms of assumptions, objectives and practical implications. PMID- 16248260 TI - [Cultural diversity and stereotyping: implication for the medical practice]. AB - Increasing number of migrants worldwide brings doctors to treat patients of various origins. Patients' diversity enriches health professionals but also induces a risk of mutual incomprehension, due to cultural and language barriers. Multicultural context stimulates unwittingly stereotyping, based on a simplistic assessment of the patient's culture. Stereotyping is also influenced by the political and media coverage. Studies underscored that universally, minorities patients have an unequal access to health care in host countries. Health professionals should be aware that racial stereotyping exists in medical practice: it is a first step to bridge cultural gap between them and their patients. PMID- 16248262 TI - [Social inequalities and health: experiences of a mobile health care unit in Geneva]. AB - Health care professionals are in contact with patients of all social horizons. Healthcare professionals come into contact with patients from all social backgrounds. Low socio-economic status is a well-known determinant of morbidity and mortality. A mobile community health care unit ("Umsco") was created in Geneva in 1996 with the aim of providing health care to very low-income patients who aren't catered for by the traditional healthcare system in Switzerland. The Mobile Unit's patients are mainly female South American illegal immigrants, but also the city' homeless. This article explains how the Unit functions on a day-to day basis. We describe socio-demographic characteristics of the patients and their main presenting complaints. We also discuss how health care professionals in general can adapt to the specific needs of socially deprived and vulnerable patients. PMID- 16248261 TI - [Sociocultural diversity and medical education]. AB - Global migration patterns have led to increasingly diverse populations, and physicians must learn to work effectively with patients from diverse backgrounds. However, some aspects of medical education may actually reinforce the idea that physicians do not need to take social and cultural differences into consideration in their work. In order to train physicians who are capable of providing quality care to patients from diverse backgrounds, it will be important to integrate the concept of cultural competence into all aspects of medical training, such that it will cease to be perceived as an "optional" aspect of medicine. PMID- 16248263 TI - [Neurosciences and neuroethics]. PMID- 16248264 TI - [Cardiovascular diseases and waist circumference]. PMID- 16248265 TI - [The curse of the Norwegians]. PMID- 16248266 TI - [Intravenous injection of epinephrine for anaphylaxis]. PMID- 16248267 TI - [Bird flu, this new menace that glides in our skies (4)]. PMID- 16248268 TI - [Hypertension and the health of the French president]. PMID- 16248269 TI - [The vagueness of academic medicine]. PMID- 16248270 TI - Overview of efficacy outcome variables for the evaluation of periodontal disease treatment. AB - Since the late 1980s, numerous new products for the prevention, diagnosis, and treatment of periodontal disease have been developed. These products have required randomized clinical trials to provide proof of efficacy and safety prior to marketing. Originally, no consensus existed in the scientific community, regulatory agencies, or industry as to the appropriate design, conduct, and methods of analyses of clinical trials in periodontics. However, in 1967, the Task Force on Design and Analysis in Dental and Oral Research was formed. This task force held three conferences to address specific issues in clinical oral health research; these meetings provided the foundation for the development of guidelines for clinical trials by the Subcommittee on Clinical Trials in Periodontics of the American Academy of Periodontology. Members of the subcommittee held an international symposium in 1996. Following this meeting, 4 working groups were convened and charged with the development of guidelines for the design and conduct of clinical trials in periodontics. These working groups developed guidelines for periodontal clinical trials in the areas of chemotherapeutic agents to slow or arrest periodontitis, products and methods for diagnosis and/or management of periodontitis, products designed to regenerate periodontal tissues, and clinical trials on endosseous dental implants. These guidelines have been published and approved or accepted by the American Academy of Periodontology and the Council on Scientific Affairs of the American Dental Association. PMID- 16248271 TI - Pocket depth reduction as an outcome measure of inflammation and soft tissue changes in periodontitis trials. AB - Randomized, controlled clinical trials are prospective studies involving human subjects that are designed to assess disease interventions by comparing end points related to patient well-being. The purpose of this article is to examine the parameter probing (or pocket) depth as a meaningful end-point for evaluating novel periodontitis therapies. Probing depth provides an estimate of the inflamed, ulcerated lesion secondary to periodontitis disease. Probing depth measurements strongly correlate with the concentrations of host inflammatory mediators implicated in the pathogenesis of periodontitis; these mediators are generated locally secondary to the etiologic biofilm. Probing depth is a well recognized and routine diagnostic parameter for periodontitis. It is easy to measure and interpret, sensitive to change with treatment, and correlates with other parameters of interest. Evidence from cohort studies indicates that increased probing depth is predictive of periodontitis events such as alveolar bone resorption and tooth loss. Clinical trials assessing probing depth as an efficacy outcome should feature standardization and calibration procedures to decrease measurement bias and variability. In addition, the analysis and presentation of probing-depth data should include the overall population, prognostic subcohorts, and consideration of clinical relevance. Probing depth is the most commonly reported outcome in periodontitis clinical trials and is a meaningful end-point for investigators and clinicians to judge the efficacy of periodontitis interventions. PMID- 16248272 TI - Commentary: clinical implications of periodontal disease assessments using probing depth and bleeding on probing to measure the status of the periodontal biofilm interface. AB - Periodontal disease has traditionally been classified based upon history and clinical presentation, but considerable heterogeneity persists within each diagnostic category. Clinical signs have relatively weak sensitivity and specificity for predicting disease outcomes in treated and untreated individuals, suggesting the need for improved periodontal classifications, prognosis, and treatment strategies. Data from an epidemiologic study that included full-mouth clinical periodontal measurements from more than 6700 community-dwelling subjects were analyzed to identify heterogeneity in disease presentation. These analyses were used to create periodontal disease classifications based upon probing depths (PD) and bleeding on probing (BOP) scores. Two distinct disease classifications emerged: i.e. moderate periodontitis (MP) and severe periodontitis (SP). MP was defined as 1 or more sites with a PD of 4 mm or greater and BOP extent scores less than 50%, and SP was defined as 1 or more sites with a PD of 4 mm or greater and BOP extent scores of 50% or higher. These definitions were used to retrospectively analyze data from a pilot study designed to determine the clinical effects of scaling and root planing (SRP) treatment. Examiners evaluated 58 subjects for potential differences in treatment response comparing MP with SP subjects at baseline and 6 weeks after SRP treatment. Plaque scores were associated with greater extent of pocketing (extent PD > or =4 mm) in the MP group, but not in the SP group. Significant clinical improvement in PD and BOP was detected at 6 weeks in the SP group but not in the MP group when using the presented classification to retrospectively compare patient response to SRP treatment using a separate clinical data set. Periodontal disease is a heterogeneous condition, and classifications based upon BOP and PD may be useful considerations in predicting patient clinical response to therapy. PMID- 16248273 TI - Clinical attachment level change as an outcome measure for therapies that slow the progression of periodontal disease. AB - Attachment levels are excellent indicators of past destruction of the periodontal attachment apparatus and can be used to monitor the progression of periodontitis. They have been used in clinical trials to monitor the efficacy of a variety of therapeutic modalities that may either slow the progression of periodontal disease or allow for regeneration of lost attachment and supporting structures. Inherent difficulties in accurately assessing attachment levels include inflammation, which causes coronal displacement of the gingival margin without a concomitant migration of the dentogingival epithelium to a level apical to the cementoenamel junction, and recessions, in which an obvious loss of attachment has occurred, but there is no increase in probing depth. Attachment level measurements are more frequently used as clinical end-points in clinical trials than by private practitioners to determine the periodontal status of patients and to monitor patient responses to periodontal therapy. Clinical attachment level measurements have been used in clinical trials to evaluate a systemic host modulatory agent, demonstrating their utility as surrogate markers of efficacy. PMID- 16248274 TI - Radiographic alveolar bone change as an outcome measure for therapies that inhibit bone loss or foster bone gain. AB - Periodontal clinical trials rely on signs of gingival inflammation, periodontal probing methods, and intra-oral radiographic methods to measure outcomes of therapies that inhibit bone loss or foster bone gain. A single radiograph can provide information on the accumulation of past disease, and sequential radiographs can determine regeneration or progression of periodontal destruction. However, the precision achieved with a radiograph is associated with the actual exposure of the film and the subsequent analysis. A film exposed using standardized geometry and standardized processing has many advantages over non standardized conventional film. If the patient population for a study is a small, selective group, or if one needs to identify the smallest amount of bone loss in the shortest time, standardized films may be used. Conversely, in a large-scale trial, the use of standardized films may prove to be impractical. Since the advent of personal computer-based image-processing workstations, digital-image analysis has also become a practical tool for clinical trials. Digital subtraction radiography is a highly precise and accurate technique for assessing bony changes associated with periodontal disease. Therefore, advanced digital imaging systems may be more cost-effective in a clinical trial because the level of precision will have an effect on study length and sample size. PMID- 16248275 TI - The clinical significance of new therapies for the management of periodontal disease. AB - With the approval of interventions or treatments for periodontal disease based on results of large clinical trials, it is often difficult for the clinician to determine whether the statistical results obtained from these studies are clinically significant and whether these interventions or treatments will thus be effective in their individual patients. In this article, a series of measures of clinical significance are derived from the literature, and a 2-step approach is suggested for determining clinical significance. First, the reliability of the data as a function of the reliability of the measurement tool(s) is calculated, and then clinical significance is put into the context of effect size. By using these estimators, clinicians can determine, for example, whether administration of an experimental local medication plus scaling and root planing provides a clinically significant outcome, which can assist them in deciding which interventions are most effective for their patients. PMID- 16248276 TI - Herpes simplex virus-based vectors for the treatment of cancer and neurodegenerative disease. AB - Vectors based on herpes simplex virus (HSV) are being developed for use in human neurodegenerative diseases and cancer. In neurodegenerative disease, this involves the use of highly disabled, non-replicating HSV vectors engineered to carry a therapeutic gene. In contrast, the use of HSV vectors in cancer involves partially disabled viruses that can replicate in dividing cells but not in non dividing cells and therefore have an oncolytic effect. Both these approaches have produced promising results in cell culture and animal models. Moreover, phase I clinical trials have demonstrated the safety of HSV vectors and their possible efficacy in otherwise untreatable cancers. PMID- 16248277 TI - Viral vectors as therapeutic agents for glioblastoma. AB - Established treatments such as surgery, radiation and chemotherapy have not altered the median survival of glioblastoma, the most common malignant brain tumor. Since these failures reflect the highly invasive nature of glioblastoma, as well as the fact that few cells are actively replicating at any given point in time, therapies need to act in areas of the brain distant from the site of tumor origin and for long after their introduction. Over the past decade, laboratory studies and early clinical trials have raised hope that these therapeutic requirements may be fulfilled by gene therapy using non-replicating transgene bearing viruses, oncolytic viruses or migratory stem cells to deliver tumoricidal transgenes. The principles behind these approaches and their initial results are reviewed. PMID- 16248278 TI - Intrathecal gene transfer by adeno-associated virus for pain. AB - Chronic pain is among the most prevalent medical problems, affecting more than half of patients with advanced cancer and many with other common diseases. Current analgesics often fail to provide satisfactory symptom relief and frequently cause severe side effects. Intrathecal (IT) gene transfer is an attractive method for pain research in rodent models, because it allows targeting of a wide variety of secretable peptides and proteins to the spinal cord, an important neural center for the processing of nociceptive signals. The potential of IT gene transfer for improving opioid therapy and for validating new analgesic targets, such as cytokines involved in spinal glial activation, is discussed. The IT space has been notoriously resistant to efficient gene transfer, limiting therapeutic gene expression to less than 2 weeks with most vector systems. Recent progress with adeno-associated virus (AAV) technology allowed efficient long-term gene expression, facilitating studies reflective of the chronic nature of many pain states. AAV is one of the most advanced gene therapy vectors currently undergoing clinical trials for a variety of disorders. In patients, AAV vectors could be administered intrathecally by a lumbar puncture, a safe procedure routinely performed at the bedside. AAV vectors may therefore become an important tool for translational studies to validate newly identified therapeutic targets in clinical pain states. PMID- 16248279 TI - Intrapleural 'outside-in' gene therapy: therapeutics for organs of the chest via gene transfer to the pleura. AB - The pleural space is an attractive site for using viral vectors to deliver gene products to the lung parenchyma, other thoracic structures and the systemic circulation. The advantages of intrapleural gene transfer using viral vectors include: (i) easy accessibility; (ii) large surface area; (iii) ability to provide high concentrations of secreted gene products to chest structures; (iv) low risk of detrimental effects of possible vector-induced inflammation compared with intravascular delivery; and (v) because it is local, lower vector doses can be used to deliver therapeutic genes to thoracic structures than less efficient systemic routes. Examples of pleural gene transfer include the use of adenovirus vectors to treat mesothelioma by transiently expressing genes that encode toxic proteins, immunomodulatory molecules or anti-angiogenesis factors. Intrapleural delivery of adeno-associated viral vectors represents an efficient strategy to treat alpha1-antitrypsin (alpha1AT) deficiency, achieving high lung and systemic therapeutic levels of alpha1AT. Intrapleural delivery of gene transfer vectors holds promise for the treatment of diseases requiring transient, localized gene expression, as well as sustained expression of genes to correct hereditary disorders requiring localized or systemic expression of the therapeutic protein. PMID- 16248281 TI - Gene therapy in the treatment of Fanconi anemia, a progressive bone marrow failure syndrome. AB - Fanconi anemia (FA) is a genetic disease characterized by progressive, fatal bone marrow failure, congenital anomalies and predisposition to cancer. Although stem cell transplantation is therapeutic, human leukocyte antigen-identical sibling donors are available to a minority of patients. In murine models and human cells in vitro, gene transfer corrects the FA cellular phenotype of chromosomal breakage in response to DNA-damaging agents, suggesting therapeutic use of gene transfer is possible. However, disease-specific characteristics make application of viral vector technology difficult. Multiple studies are currently underway to develop a gene therapy approach for treating this disease, including phase I trials. PMID- 16248280 TI - Listeria-based vaccines for cancer treatment. AB - Listeria monocytogenes is a facultative intracellular bacterium that enters professional antigen-presenting cells by active phagocytosis. As a live bacterium, it induces antigen-presenting cell maturation and strong innate immunity which may assist in the immune response to poorly immunogenic antigens, such as tumor-associated antigens. Listeria produces virulence factors that allow it to escape from the phagolysosome and colonize the cytosol of the host cell. It is thus a potent vaccine vector for the presentation of passenger antigens to the major histocompatibility complex class I and II pathways of antigen processing and presentation. Recent progress in developing this bacterium as a vaccine vector for tumor-associated antigens is reviewed. In mouse models, recombinant Listeria carrying a number of such antigens has provided therapeutic immunity directed towards established tumors. Safety issues associated with live bacterial vaccine vectors and problems to be overcome in developing Listeria as a cancer immunotherapeutic for human use are also discussed. PMID- 16248282 TI - Gene therapy for type 1 diabetes: a proposal to move to the next level. AB - A number of studies have confirmed that gene therapy can be successfully applied to prevent type 1 diabetes mellitus and to facilitate the transplantation of replacement insulin-producing cells as allografts of the islets of Langerhans or in the form of enriched and homogeneous surrogate cells. However, these successes have remained restricted to a few rodent models and have not progressed past this stage into larger animals. Newer technology has also emerged that offers exciting alternatives or complementary methods to achieve the same results, including therapeutic cloning. A synopsis of the strategies and tools used successfully in vivo is provided and a proposal to motivate a step-up of these approaches to larger animals, if not outright to safety trials in humans, is outlined. Additionally, newer emerging technologies that can, in the future, complement those that are currently readily adaptable to clinical implementation are suggested. PMID- 16248284 TI - Technology evaluation: TransMID, KS Biomedix/Nycomed/Sosei/PharmaEngine. AB - KS Biomedix (formerly Avicenna Medica; now a subsidiary of the Xenova group) and Nycomed, together with Japanese licensee Sosei and Chinese licensee PharmaEngine, are developing TransMID, a transferrin-mediated diphtheria toxin delivery system for the potential treatment of adult, recurrent, inoperable, high-grade glioma (as TransMID-107R). It is also under investigation for other forms of brain cancer, including early brain cancer (as TransMID-107N), metastatic brain cancer (as TransMID-107M) and pediatric brain cancer (as TransMID-107P). TransMID is currently undergoing phase III clinical trials. PMID- 16248283 TI - Making antisense of splicing. AB - Alternative splicing multiplies the coding capacity of the genome, resulting in an expanded proteome that provides many targets for therapy. In addition to creating this diverse pharmacoproteome, the process of splicing can be targeted by conventional and molecular therapies. Splicing as a therapeutic target is highlighted in this review, with a particular emphasis on oligonucleotide-based molecular approaches. These oligonucleotides can be used to promote skipping of constitutive exons, inhibit inappropriately activated exons, or stimulate exons weakened by mutations. Preliminary, but exciting, results suggest that these reagents could have clinical utility in treating previously intractable conditions. PMID- 16248285 TI - Technology evaluation: VEGF Trap (cancer), Regeneron/sanofi-aventis. AB - sanofi-aventis (formerly Aventis) and Regeneron are developing systemic VEGF Trap, a soluble decoy receptor comprising portions of VEGF receptors 1 and 2, for the potential intravenous/subcutaneous treatment of cancer. PMID- 16248286 TI - Technology evaluation: mapatumumab, Human Genome Sciences/GlaxoSmithKline/Takeda. AB - Human Genome Sciences and GlaxoSmithKline, under license from Cambridge Antibody Technology, are developing mapatumumab, the intravenously administered lead from a series of human monoclonal antibody tumor necrosis factor-related apoptosis inducing ligand receptor 1 agonists, for the potential treatment of cancer. The therapy is currently undergoing phase II clinical trials. Takeda is developing mapatumumab in Japan. PMID- 16248287 TI - Technology evaluation: Onyvax-P, Onyvax. AB - Onyvax is developing Onyvax-P, an allogeneic whole-cell vaccine, for the potential treatment of prostate cancer. The vaccine is currently undergoing phase II clinical trials. PMID- 16248288 TI - Lifespan differences in the social networks of prison inmates. AB - Socioemotional Selectivity Theory (SST) (Carstensen, 1992, 1993) accounts for lifespan changes in human social networks and for the motivations which underlie those changes. SST is applied in this research with 256 prison inmates and non inmates, ages 18-84, from Mississippi, Kansas, and New Mexico. Two research questions sought to identify (a) whether inmate networks change in size, and (b) whether overall closeness within an inmate's network changes over the adult years. Results indicate that older inmates, much like older non-inmates, have few peripheral partners, are buffered from the wider population of prisoners, and interact within a small group of very close partners. Although older inmates are not completely isolated, they do maintain fewer network partners as age increases, like their non-incarcerated counterparts, and overall are as emotionally close to network members as non-inmates. PMID- 16248290 TI - Effects of parental rejection and relationship quality on depression among older rural adults. AB - Research suggests that families have development histories which influence their responses to situations across the life course. The present study examined the effects of family relationship histories on intergenerational affection and conflict between adult children and their parents and how this affects parents' depressive symptoms. Path analysis based on matched reports of adult children and their parents revealed that parental behavior early in life affects contemporary relationships between family members, which ultimately impacts parents' depressive symptoms. Parental rejection is negatively associated with intergenerational affection, which in turn is negatively associated with depressive symptoms. PMID- 16248289 TI - Social, health, and age differences associated with depressive disorders in women with rheumatoid arthritis. AB - Depression in women with rheumatoid arthritis (RA) may be related to social role experiences, physical health, and age. The purpose of this study was to examine the social and health factors contributing to depression in two age groups of women with RA. One-hundred and thirty-eight midlife and late-life women with a diagnosis of RA participated in this cross sectional survey study. Multiple regression analysis indicated that social role balance, functional status, number of co-existing health problems, and age were significant predictors of depression in midlife and late-life women with RA. Role balance was the strongest factor contributing to a woman's depression score. Compared to midlife women, late-life women reported significantly higher role balance and lower depression scores, despite poorer functional status and more concomitant health problems. PMID- 16248291 TI - "At this point now": older workers' reflections on their current employment experiences. AB - Despite the frequently referenced graying of the U.S. workforce, we know relatively little about the work-related concerns and experiences of older workers--those aged 55+. This qualitative study addresses that gap by examining the current employment situations of a purposive sample of 37 older workers. Thematic content analysis revealed a vast diversity in the concerns and experiences of contemporary older workers, with participants being actively involved in all of the traditional stages of work-life development. Findings suggest a deepened or more nuanced view of the principal meanings of work--in particular, identity and social interaction--and highlight several negative aspects of later-life employment such as pension insecurity, unemployment, age discrimination, not having found one's "niche," and chronic interpersonal difficulties with co-workers. Findings are discussed against the backdrop of a rapidly changing workplace and dramatically shifting work-retirement trajectories in which job stability and predictable retirement are less common. PMID- 16248293 TI - PCT upheaval could kill clinical engagement, PEC chairs warn. PMID- 16248292 TI - Difficulties, disagreements, and disappointments in late-life marriages. AB - Although research has examined marital satisfaction in later life, little is known about the specific relationship challenges that older couples experience. Thus, the objective of the study was to explore the challenges older couples face. Data came from the USC Longitudinal Study of Generations. Qualitative analysis was conducted on 105 older couples, with an average age of 69, who responded in 1997 to an open-ended question about difficulties, disagreements, and disappointments in their marriage. Ten themes emerged from the data. In order of frequency, they were leisure activities, intimacy, finances, no problems, personality, intergenerational relations, household concerns, personal habits, health issues, and work/retirement. Comparisons in responses were made for age, gender, marital quality, length of marriage, and health. In addition, the dyadic data were examined for patterns within the relationship. PMID- 16248295 TI - Hospital closures a 'red herring'. PMID- 16248294 TI - 'Information is power' say patient. PMID- 16248296 TI - Patient choice. The big sell: why it's time for hospitals to market themselves. PMID- 16248297 TI - On public expectations. PMID- 16248299 TI - Clinical management. Fit for purpose. AB - Care for people with epilepsy has been dogged by lack of services and long waiting lusts. There have been some pockets of improvement, aided by GPs with speciaI interests and networks of specialists. Experts believe GPs will have a key role in providing better services for epilepsy sufferers. PMID- 16248298 TI - Barometer. Acute trusts September 2005. PMID- 16248300 TI - Good management. A sporting chance. PMID- 16248301 TI - Development and validation of a CO-C7 FE complex for biomechanical study. AB - In this study, the digitized geometrical data of the embalmed skull and vertebrae (C0-C7) of a 68-year old male cadaver were processed to develop a comprehensive, geometrically accurate, nonlinear C0-C7 FE model. The biomechanical response of human neck under physiological static loadings, near vertex drop impact and rear end impact (whiplash) conditions were investigated and compared with published experimental results. Under static loading conditions, the predicted moment rotation relationships of each motion segment under moments in midsagittal plane and horizontal plane agreed well with experimental data. In addition, the respective predicted head impact force history and the S-shaped kinematics responses of head-neck complex under near-vertex drop impact and rear-end conditions were close to those observed in reported experiments. Although the predicted responses of the head-neck complex under any specific condition cannot perfectly match the experimental observations, the model reasonably reflected the rotation distributions among the motion segments under static moments and basic responses of head and neck under dynamic loadings. The current model may offer potentials to effectively reflect the behavior of human cervical spine suitable for further biomechanics and traumatic studies. PMID- 16248302 TI - Determining effective centroid position in biomechanical testing: a technique for simplifying whole bone analysis. AB - BACKGROUND: Whole bone in vitro biomechanical compressive testing can be complicated by three factors: sample asymmetry, heterogeneous material properties, and unknown effective centroid location. METHOD OF APPROACH: The technique presented here facilitates the calculation of effective centroid position, modulus of elasticity and equivalent uniform strain magnitude for a cross section of bone from a simple whole bone compressive test. Simplification of section response to load is achieved through a combination of linear beam and simple planer geometry theory. The technique requires three longitudinal strain gauges be affixed around the test specimen cross section of interest, gauge position need not be determined. Sample loading is then accomplished using a simple four point loading jig. RESULTS: Results of the technique are presented on an object with known elasticity and geometry, an aluminium tube, and seven pairs of equine third metacarpal whole bones. CONCLUSIONS: Average cross section modulus of elasticity, equivalent uniform cross section strain, and effective centroid locations were all predicted to within the range of published values. Employing the testing setup and analysis technique presented in this paper resulted in a significant savings in both implementation complexity and cost over previously available techniques. PMID- 16248303 TI - The development of structural and mechanical anisotropy in fibroblast populated collagen gels. AB - An in vitro model system was developed to study structure-function relationships and the development of structural and mechanical anisotropy in collagenous tissues. Fibroblast-populated collagen gels were constrained either biaxially or uniaxially. Gel remodeling, biaxial mechanical properties, and collagen orientation were determined after 72 h of culture. Collagen gels contracted spontaneously in the unconstrained direction, uniaxial mechanical constraints produced structural anisotropy, and this structural anisotropy was associated with mechanical anisotropy. Cardiac and tendon fibroblasts were compared to test the hypothesis that tendon fibroblasts should generate greater anisotropy in vitro. However, no differences were seen in either structure or mechanics of collagen gels populated with these two cell types, or between fibroblast populated gels and acellular gels. This study demonstrates our ability to control and measure the development of structural and mechanical anisotropy due to imposed mechanical constraints in a fibroblast-populated collagen gel model system. While imposed constraints were required for the development of anisotropy in this system, active remodeling of the gel by fibroblasts was not. This model system will provide a basis for investigating structure-function relationships in engineered constructs and for studying mechanisms underlying the development of anisotropy in collagenous tissues. PMID- 16248304 TI - Correlation of HSP70 expression and cell viability following thermal stimulation of bovine aortic endothelial cells. AB - Thermal preconditioning protocols for cardiac cells were identified which produce elevated HSP70 levels while maintaining high cell viability. Bovine aortic endothelial cells were heated with a water bath at temperatures ranging from 44 to 50 degrees C for periods of 1-30 min. Thermal stimulation protocols were determined which induce HSP70 expression levels ranging from 2.3 to 3.6 times the control while maintaining cell viabilities greater than 90%. An Arrhenius injury model fit to the cell damage data yielded values of A = 1.4 X 10(66) s(-1) and Ea = 4.1 X 10(5) J/mol. Knowledge of the injury parameters and HSP70 kinetics will enhance dosimetry guideline development for thermal stimulation of heat shock proteins expression in cardiac tissue. PMID- 16248305 TI - Nutrient utilization by bovine articular chondrocytes: a combined experimental and theoretical approach. AB - A combined experimental-numerical approach was adopted to characterize glucose and oxygen uptake and lactate production by bovine articular chondrocytes in a model system. For a wide range of cell concentrations, cells in agarose were supplemented with either low or high glucose medium. During an initial culture phase of 48 h, oxygen was monitored noninvasively using a biosensor system. Glucose and lactate were determined by medium sampling. In order to quantify glucose and oxygen uptake, a finite element approach was adopted to describe diffusion and uptake in the experimental model. Numerical predictions of lactate, based on simple relations for cell metabolism, were found to agree well for low glucose, but not for high glucose medium. Oxygen did not play a role in either case. Given the close association between chondrocyte energy metabolism and matrix synthesis, a quantifiable prediction of utilization can present a valuable contribution in the optimization of tissue engineering conditions. PMID- 16248306 TI - 3-D numerical simulation of blood flow through models of the human aorta. AB - A Spiral Computerized Tomography (CT) scan of the aorta were obtained from a single subject and three model variations were examined. Computational fluid dynamics modeling of all three models showed variations in the velocity contours along the aortic arch with differences in the boundary layer growth and recirculation regions. Further down-stream, all three models showed very similar velocity profiles during maximum velocity with differences occurring in the decelerating part of the pulse. Flow patterns obtained from transient 3-D computational fluid dynamics are influenced by different reconstruction methods and the pulsatility of the flow. Caution is required when analyzing models based on CT scans. PMID- 16248308 TI - Flow in a mechanical bileaflet heart valve at laminar and near-peak systole flow rates: CFD simulations and experiments. AB - Time-accurate, fully 3D numerical simulations and particle image velocity laboratory experiments are carried out for flow through a fully open bileaflet mechanical heart valve under steady (nonpulsatile) inflow conditions. Flows at two different Reynolds numbers, one in the laminar regime and the other turbulent (near-peak systole flow rate), are investigated. A direct numerical simulation is carried out for the laminar flow case while the turbulent flow is investigated with two different unsteady statistical turbulence modeling approaches, unsteady Reynolds-averaged Navier-Stokes (URANS) and detached-eddy simulation (DES) approach. For both the laminar and turbulent cases the computed mean velocity profiles are in good overall agreement with the measurements. For the turbulent simulations, however, the comparisons with the measurements demonstrate clearly the superiority of the DES approach and underscore its potential as a powerful modeling tool of cardiovascular flows at physiological conditions. The study reveals numerous previously unknown features of the flow. PMID- 16248307 TI - Thin-film coupled fluid-solid analysis of flow through the Ahmed glaucoma drainage device. AB - The Ahmed glaucoma valve (AGV) is a popular glaucoma drainage device, allowing maintenance of normal intraocular pressure in patients with reduced trabecular outflow facility. The uniquely attractive feature of the AGV, in contrast to other available drainage devices, is its variable resistance in response to changes in flow rate. As a result of this variable resistance, the AGV maintains a pressure drop between 7 and 12 mm Hg for a wide range of aqueous humor flow rates. In this paper, we demonstrate that the nonlinear behavior of the AGV is a direct result of the flexibility of the valve material. Due to the thin geometry of the system, the leaflets of the AGV were modeled using the von Karman plate theory coupled to a Reynolds lubrication theory model of the aqueous humor flow through the valve. The resulting two-dimensional coupled steady-state partial differential equation system was solved by the finite element method. The Poisson's ratio of the valve was set to 0.45, and the modulus was regressed to experimental data, giving a best-fit value 4.2 MPa. Simulation results compared favorably with previous experimental studies and our own pressure-drop/flow-rate data. For an in vitro flow of 1.6 microL/min, we calculated a pressure drop of 5.8 mm Hg and measured a pressure drop of 5.2 +/- 0.4 mm Hg. As flow rate was increased, pressure drop rose in a strongly sublinear fashion, with a flow rate of 20 microL/min giving a predicted pressure drop of only 10.9 mm Hg and a measured pressure drop of 10.5 +/- 1.1 mm Hg. The AGV model was then applied to simulate in vivo conditions. For an aqueous humor flow rate of 1.5-3.0 microL/min, the calculated pressure drops were 5.3 and 6.3 mm Hg. PMID- 16248310 TI - An intelligent control method based on fuzzy logic for a robotic testing system for the human spine. AB - In previous biomechanical studies of the human spine, we implemented a hybrid controller to investigate load-displacement characteristics. We found that measurement errors in both position and force caused the controller to be less accurate than predicted. As an alternative to hybrid control, a fuzzy logic controller (FLC) has been developed and implemented in a robotic testing system for the human spine. An FLC is a real-time expert system that can emulate part of a human operator's knowledge by using a set of action rules. The FLC provides simple but robust solutions that cover a wide range of system parameters and can cope with significant disturbances. It can be viewed as a heuristic and modular way of defining a nonlinear, table-based control system. In this study, an FLC is developed which uses the force difference and the change in force difference as the input parameters, and the displacement as the output parameter. A rule-table based on these parameters is designed for the controller Experiments on a physical model composed of springs demonstrate the improved performance of the proposed method. PMID- 16248309 TI - Effect of gravity on liquid plug transport through an airway bifurcation model. AB - Many medical therapies require liquid plugs to be instilled into and delivered throughout the pulmonary airways. Improving these treatments requires a better understanding of how liquid distributes throughout these airways. In this study, gravitational and surface mechanisms determining the distribution of instilled liquids are examined experimentally using a bench-top model of a symmetrically bifurcating airway. A liquid plug was instilled into the parent tube and driven through the bifurcation by a syringe pump. The effect of gravity was adjusted by changing the roll angle (phi) and pitch angle (gamma) of the bifurcation (phi = gamma =0 deg was isogravitational). Phi determines the relative gravitational orientation of the two daughter tubes: when phi not equal to 0 deg, one daughter tube was lower (gravitationally favored) compared to the other. Gamma determines the component of gravity acting along the axial direction of the parent tube: when gamma not equal to 0 deg, a nonzero component of gravity acts along the axial direction of the parent tube. A splitting ratio Rs, is defined as the ratio of the liquid volume in the upper daughter to the lower just after plug splitting. We measured the splitting ratio, Rs, as a function of: the parent-tube capillary number (Cap); the Bond number (Bo); phi; gamma; and the presence of pre existing plugs initially blocking either daughter tube. A critical capillary number (Cac) was found to exist below which no liquid entered the upper daughter (Rs = 0), and above which Rs increased and leveled off with Cap. Cac increased while Rs decreased with increasing phi, gamma, and Bo for blocked and unblocked cases at a given Cap > Ca,. Compared to the nonblockage cases, Rs decreased (increased) at a given Cap while Cac increased (decreased) with an upper (lower) liquid blockage. More liquid entered the unblocked daughter with a blockage in one daughter tube, and this effect was larger with larger gravity effect. A simple theoretical model that predicts Rs and Cac is in qualitative agreement with the experiments over a wide range of parameters. PMID- 16248312 TI - Pre-strained epimuscular connections cause muscular myofascial force transmission to affect properties of synergistic EHL and EDL muscles of the rat. AB - BACKGROUND: Myofascial force transmission occurs between muscles (intermuscular myofascial force transmission) and from muscles to surrounding nonmuscular structures such as neurovascular tracts and bone (extramuscular myofascial force transmission). The purpose was to investigate the mechanical role of the epimuscular connections (the integral system of inter- and extramuscular connections) as well as the isolated role of extramuscular connections on myofascial force transmission and to test the hypothesis, if such connections are prestrained. METHOD OF APPROACH: Length-force characteristics of extensor hallucis longus (EHL) muscle of the rat were measured in two conditions: (I) with the neighboring EDL muscle and epimuscular connections of the muscles intact: EDL was kept at a constant muscle tendon complex length. (II) After removing EDL, leaving EHL with intact extramuscular connections exclusively. RESULTS: (I) Epimuscular connections of the tested muscles proved to be prestrained significantly. (1) Passive EHL force was nonzero for all isometric EHL lengths including very low lengths, increasing with length to approximately 13% of optimum force at high length. (2) Significant proximodistal EDL force differences were found at all EHL lengths: Initially, proximal EDL force = 1.18 +/- 0.11 N, where as distal EDL force = 1.50 +/- 0.08 N (mean +/- SE). EHL lengthening decreased the proximo-distal EDL force difference significantly (by 18.4%) but the dominance of EDL distal force remained. This shows that EHL lengthening reduces the prestrain on epimuscular connections via intermuscular connections; however; the prestrain on the extramuscular connections of EDL remains effective. (II) Removing EDL muscle affected EHL forces significantly. (1) Passive EHL forces decreased at all muscle lengths by approximately 17%. However, EHL passive force was still non-zero for the entire isometric EHL length range, indicating pre-strain of extramuscular connections of EHL. This indicates that a substantial part of the effects originates solely from the extramuscular connections of EHL. However, a role for intermuscular connections between EHL and EDL, when present, cannot be excluded. (2) Total EHL forces included significant shape changes in the length-force curve (e.g., optimal EHL force decreased significantly by 6%) showing that due to myofascial force transmission muscle length-force characteristics are not specific properties of individual muscles. CONCLUSIONS: The pre-strain in the epimuscular connections of EDL and EHL indicate that these myofascial pathways are sufficiently stiff to transmit force even after small changes in relative position of a muscle with respect to its neighboring muscular and nonmuscular tissues. This suggests the likelihood of such effects also in vivo. PMID- 16248311 TI - Comparison of deformable and elastic foundation finite element simulations for predicting knee replacement mechanics. AB - Rigid body total knee replacement (TKR) models with tibiofemoral contact based on elastic foundation (EF) theory utilize simple contact pressure-surface overclosure relationships to estimate joint mechanics, and require significantly less computational time than corresponding deformable finite element (FE) methods. However, potential differences in predicted kinematics between these representations are currently not well understood, and it is unclear if the estimates of contact area and pressure are acceptable. Therefore, the objectives of the current study were to develop rigid EF and deformable FE models of tibiofemoral contact, and to compare predicted kinematics and contact mechanics from both representations during gait loading conditions with three different implant designs. Linear and nonlinear contact pressure-surface overclosure relationships based on polyethylene material properties were developed using EF theory. All other variables being equal, rigid body FE models accurately estimated kinematics predicted by fully deformable FE models and required only 2% of the analysis time. As expected, the linear EF contact model sufficiently approximated trends for peak contact pressures, but overestimated the deformable results by up to 30%. The nonlinear EF contact model more accurately reproduced trends and magnitudes of the deformable analysis, with maximum differences of approximately 15% at the peak pressures during the gait cycle. All contact area predictions agreed in trend and magnitude. Using rigid models, edge-loading conditions resulted in substantial overestimation of peak pressure. Optimal nonlinear EF contact relationships were developed for specific TKR designs for use in parametric or repetitive analyses where computational time is paramount. The explicit FE analysis method utilized here provides a unique approach in that both rigid and deformable analyses can be run from the same input file, thus enabling simple selection of the most appropriate representation for the analysis of interest. PMID- 16248313 TI - A method for measurement of joint kinematics in vivo by registration of 3-D geometric models with cine phase contrast magnetic resonance imaging data. AB - A new method is presented for measuring joint kinematics by optimally matching modeled trajectories of geometric surface models of bones with cine phase contrast (cine-PC) magnetic resonance imaging data. The incorporation of the geometric bone models (GBMs) allows computation of kinematics based on coordinate systems placed relative to full 3-D anatomy, as well as quantification of changes in articular contact locations and relative velocities during dynamic motion. These capabilities are additional to those of cine-PC based techniques that have been used previously to measure joint kinematics during activity. Cine-PC magnitude and velocity data are collected on a fixed image plane prescribed through a repetitively moved skeletal joint. The intersection of each GBM with a simulated image plane is calculated as the model moves along a computed trajectory, and cine-PC velocity data are sampled from the regions of the velocity images within the area of this intersection. From the sampled velocity data, the instantaneous linear and angular velocities of a coordinate system fixed to the GBM are estimated, and integration of the linear and angular velocities is used to predict updated trajectories. A moving validation phantom that produces motions and velocity data similar to those observed in an experiment on human knee kinematics was designed. This phantom was used to assess cine-PC rigid body tracking performance by comparing the kinematics of the phantom measured by this method to similar measurements made using a magnetic tracking system. Average differences between the two methods were measured as 2.82 mm rms for anterior/posterior tibial position, and 2.63 deg rms for axial rotation. An intertrial repeatability study of human knee kinematics using the new method produced rms differences in anterior/posterior tibial position and axial rotation of 1.44 mm and 2.35 deg. The performance of the method is concluded to be sufficient for the effective study of kinematic changes caused to knees by soft tissue injuries. PMID- 16248314 TI - Experimental investigation into the deep penetration of soft solids by sharp and blunt punches, with application to the piercing of skin. AB - An experimental study has been conducted on the penetration of silicone rubbers and human skin in vivo by sharp-tipped and flat-bottomed cylindrical punches. A penetrometer was developed to measure the penetration of human skin in vivo, while a conventional screw-driven testing machine was used to penetrate the silicone rubbers. The experiments reveal that the penetration mechanism of a soft solid depends upon the punch tip geometry: a sharp tipped punch penetrates by the formation and wedging open of a mode I planar crack, while a flat-bottomed punch penetrates by the growth of a mode II ring crack. The planar crack advances with the punch, and friction along the flanks of the punch leads to a rising load versus displacement response. In contrast, the flat-bottomed punch penetrates by jerky crack advance and the load on the punch is unsteady. The average penetration pressure on the shank cross section of a flat-bottomed punch exceeds that for a sharp-tipped punch of the same diameter In addition, the penetration pressure decreases as the diameter of the sharp-tipped punch increases. These findings are in broad agreement with the predictions of Shergold and Fleck [Proc. R. Soc. London, Ser. A (in press)] who proposed models for the penetration of a soft solid by a sharp-tipped and flat-bottomed punch. PMID- 16248315 TI - Estimation of vibration power absorption density in human fingers. AB - The absorption of hand-transmitted vibration energy may be an etiological factor in vibration-induced disorders. The vibration power absorption density (VPAD) may be a better measure of energy than the total power absorption of the hand-arm system. The objectives of the present study are to develop a method to estimate the average absorption density in the fingers and to investigate its basic characteristics. Ten healthy male subjects were used in this study. The biodynamic response of the fingers in a power grip subjected to a broad-band random excitation was measured under three grip forces (15, 30, 50 N) and three push forces (35, 45, 50 N). The response was used to estimate the total finger energy absorption. The response, together with the finger volume, was also used to estimate the amount of tissue effectively involved in the absorption. Then, the average VPAD under constant-acceleration, constant-power density, constant velocity vibration spectra, and 20 tool vibration spectra were calculated. The correlations between the VPAD and the unweighted and weighted accelerations (ISO 5349-1, 2001) were also examined. The VPAD depends on both the characteristics of the vibration spectrum and the biodynamic response of the finger-hand-arm system. The biodynamic response generally plays a more important role in determining the VPAD in the middle frequency range (31.5-400 Hz) than those at the low and high ends. The applied force significantly affected the VPAD. The finger VPAD was highly correlated to the unweighted acceleration. The average VPAD can be determined using the proposed experimental method. It can serve as an alternative tool to quantify the severity of the vibration exposure for studying vibration induced finger disorders. PMID- 16248316 TI - Design of a sterile organ culture system for the ex vivo study of aortic heart valves. AB - The biological response of valves to mechanical forces is not well understood. The aim of this study was to design a pulsatile system to enable the ex vivo study of aortic valves when subjected to various hemodynamic conditions. A bioreactor was designed to subject porcine aortic valves to physiological and pathophysiological pressure and flow conditions, while maintaining viability and sterility. Pressure and flow rate could be independently controlled to produce clinically relevant mechanical conditions. The oxygen transfer rate was characterized and sterile operation was achieved over 96 hours. The oxygenation capabilities ensure sufficient oxygen transport to valves, allowing operation for extended periods. PMID- 16248317 TI - Wave intensity analysis of left ventricular filling. AB - Wave intensity analysis (WIA) is a powerful technique to study pressure and flow velocity waves in the time domain in vascular networks. The method is based on the analysis of energy transported by the wave through computation of the wave intensity dI = dPdU, where dP and dU denote pressure and flow velocity changes per time interval, respectively. In this study we propose an analytical modification to the WIA so that it can be used to study waves in conditions of time varying elastic properties, such as the left ventricle (LV) during diastole. The approach is first analytically elaborated for a one-dimensional elastic tube model of the left ventricle with a time-dependent pressure-area relationship. Data obtained with a validated quasi-three dimensional axi-symmetrical model of the left ventricle are employed to demonstrate this new approach. Along the base apex axis close to the base wave intensity curves are obtained, both using the standard method and the newly proposed modified method. The main difference between the standard and modified wave intensity pattern occurs immediately after the opening of the mitral valve. Where the standard WIA shows a backward expansion wave, the modified analysis shows a forward compression wave. The proposed modification needs to be taken into account when studying left ventricular relaxation, as it affects the wave type. PMID- 16248318 TI - Automated methodology for determination of stress distribution in human abdominal aortic aneurysm. AB - Knowledge of impending abdominal aortic aneurysm (AAA) rupture can help in surgical planning. Typically, aneurysm diameter is used as the indicator of rupture, but recent studies have hypothesized that pressure-induced biomechanical stress may be a better predictor Verification of this hypothesis on a large study population with ruptured and unruptured AAA is vital if stress is to be reliably used as a clinical prognosticator for AAA rupture risk. We have developed an automated algorithm to calculate the peak stress in patient-specific AAA models. The algorithm contains a mesh refinement module, finite element analysis module, and a postprocessing visualization module. Several aspects of the methodology used are an improvement over past reported approaches. The entire analysis may be run from a single command and is completed in less than 1 h with the peak wall stress recorded for statistical analysis. We have used our algorithm for stress analysis of numerous ruptured and unruptured AAA models and report some of our results here. By current estimates, peak stress in the aortic wall appears to be a better predictor of rupture than AAA diameter. Further use of our algorithm is ongoing on larger study populations to convincingly verify these findings. PMID- 16248319 TI - Preclinical testing of femoral hip components: an experimental investigation with four prostheses. AB - Existing standards for the preclinical testing of femoral hip implants have been successful in the objective of guaranteeing the implant's fatigue strength. There is a need for an experimental test which could ensure prostheses were not susceptible to aseptic loosening. In this study we measure the relative movement between the prosthesis and the bone of four different cemented femoral component designs in in vitro tests. The aim is to determine if differences can be distinguished and whether the differences correlate with clinical performance. The four designs are the Charnley (DePuy International Ltd., UK), the Exeter (Stryker Osteonics Howmedica Corp., USA), the Lubinus SPII (Waldemar-Link GmbH, Germany), and the Miiller Curved (JRI Ltd, UK). Five tests were carried out for each femoral component type, giving a total of 20 tests, and their permanent relative displacement (termed migration) and recoverable (i.e., elastic) relative displacement (termed inducible displacement) monitored over one million loading cycles. Considerable variation occurred in the tests. Nonetheless, most femoral components migrated medially, posteriorly, and distally. Most also rotated into varus. Translations of the Charnley (64 microns) and Lubinus (67 microns) implants were less than the Muller (72 microns) and Exeter (94 microns) implants, but this difference is not statistically significant. Most of the femoral components had rapid early migration followed by slower steady-state migration. With regard to the steady state inducible displacements of the prostheses, those of the Charnley, Exeter, and Lubinus decreased or were stable with respect to time, whilst those of the Muller typically increased with respect to time. It is concluded that migration is not a suitable basis for in vitro comparison of prosthesis designs. However inducible displacement trends provide a clinically comparable performance ranking. PMID- 16248320 TI - That hemodynamics and not material mismatch is of primary concern in bypass graft failure: an experimental argument. AB - The long term patency of end-to-side peripheral artery bypasses are low due to failure of the graft generally at the distal end of the bypass. Both material mismatch between the graft and the host artery and junction hemodynamics are cited as being major factors in disease formation at the junction. This study uses experimental methods to investigate the major differences in fluid dynamics and wall mechanics at the proximal and distal ends for rigid and compliant bypass grafts. Injection moulding was used to produce idealized transparent and compliant models of the graft/ artery junction configuration. An ePTFE graft was then used to stiffen one of the models. These models were then investigated using two-dimensional video extensometry and one-dimensional laser Doppler anemometry to determine the junction deformations and fluid velocity profiles for the rigid and complaint graft anastomotic junctions. Junction strains were evaluated and generally found to be under 5% with a peak stain measured in the stiff graft model junction of 8.3% at 100 mmHg applied pressure. Hemodynamic results were found to yield up to 40% difference in fluid velocities for the stiff/compliant comparison but up to 80% for the proximal/distal end comparisons. Similar strain conditions were assumed for the proximal and distal models while significant differences were noted in their associated hemodynamic changes. In contrasting the fluid dynamics and wall mechanics for the proximal and distal anastomoses, it is evident from the results of this study, that junction hemodynamics are the more variable factor. PMID- 16248322 TI - Fragment memories mark the end of childhood amnesia. AB - Adults described and dated two kinds of first remembrances: a personal event memory (the recollection of a personal episode that had occurred at some time in some place) and a memory fragment (an isolated memory moment having no event context and remembered, perhaps, as an image, a behavior, or an emotion). First fragment memories were judged to have originated substantially earlier in life than first event memories--approximately 3 1/3 years of age for first fragment memories versus roughly 4 years of age for first event memories. We conclude that the end of childhood amnesia is marked not by our earliest episodic memories, but by the earliest remembered fragments of childhood experiences. PMID- 16248321 TI - Migration of radio-opaque markers injected into tendon grafts: a study using roentgen stereophotogrammetric analysis (RSA). AB - An increase in anterior laxity following reconstruction of the anterior cruciate ligament (ACL) can result from lengthening of the graft construct either at the sites of fixation and/or between the sites of fixation (i.e., graft substance). Roentgen stereophotogrammetric analysis (RSA), which requires that radio-opaque markers be attached to the graft, has been shown to be a useful technique in determining lengthening in these regions. Previous methods have been used for attaching radio-opaque markers to the graft, but they all have limitations particularly for single-loop grafts. Therefore, the objective of this study was to evaluate injecting markers directly into the substance of a tendon as a viable method for measuring lengthening of single-loop graft constructs by determining the maximum amount of migration after cyclic loading. Tantalum spheres of 0.8 mm diameter were used as tendon markers. Ten single-loop tendon grafts were passed through tibial tunnels drilled in calf tibias and fixed with a tibial fixation device. Two tendon markers were inserted in one tendon bundle of each graft and the grafts were cyclically loaded for 225,000 cycles from 20 N to 170 N. At specified intervals, simultaneous radiographs were obtained of the tendon markers. Marker migration was computed as the change in distance between the two tendon markers parallel to the axis of the tibial tunnel. Marker migration had a root mean square (RMS) value of less than 0.1 mm. Because the RMS value indicates the error introduced into measurements of lengthening and because this error is negligible, the method described for attaching markers to single-loop ACL grafts has the potential to be useful for determining lengthening of single-loop ACL graft constructs in in vivo studies in humans. PMID- 16248323 TI - Directed forgetting of autobiographical events. AB - Two diary experiments demonstrated directed forgetting (DF) of autobiographical events, previously observed only for less complex memory items. Using a 2-week diary paradigm, we compared recall between a group of participants who were directed to forget Week 1 memories (forget group) and a group who did not receive a forget instruction (remember group). In Experiment 1, the forget group remembered fewer items from Week 1 than did the remember group. The effect was observed for negative and positive valence events, as well as for high and low emotional intensity events. The effect was replicated in Experiment 2 despite a memorable holiday (Valentine's Day) that occurred during the manipulation week. Forget participants remembered fewer low emotional intensity items in Experiment 2. We conclude that intentional forgetting is a plausible explanation for the loss of some autobiographical memories. PMID- 16248324 TI - Recall order determines the magnitude of directed forgetting in the within participants list method. AB - In three experiments, we investigated the effect of recall order on directed forgetting when the within-participants list method is used. Experiment 1 showed that participants tend to recall to-be-remembered (R) items before to-be forgotten (F) items when they can recall items in any order. In Experiment 2, recall order was manipulated (F-R or R-F). The results showed that only the R-F order led to directed forgetting. Finally, in Experiment 3, recall order was also manipulated, and half of the participants were explicitly instructed to use a relational strategy when both F and R items were presented. Again, only the R-F order led to directed forgetting. These results demonstrate that directed forgetting under the list method hinges on the output order in which participants recall the F and R information. Thus, output order should be taken into account by researchers investigating specific mechanisms that lead to directed forgetting. PMID- 16248325 TI - Interaction of task readiness and automatic retrieval in task switching: negative priming and competitor priming. AB - When subjects switch between tasks, performance is slower after a task switch than after a task repetition, even when preparation time is long. We report two experiments that support the idea that a large part of these residual task shift costs can be due to stimulus-cued retrieval of previous task episodes. We demonstrate that there are two different factors at work: (1) facilitation of response to the current distractor stimulus, appropriate to the previously relevant, competing task (competitor priming), and (2) impaired processing of previously suppressed responses (negative priming). Negative priming was contingent on the size of the stimulus set, suggesting that distractor suppression comes into effect only if the distractors are highly activated. Importantly, both types of interference interacted with task readiness: Whereas in the nondominant task (picture naming), switch and nonswitch trials were equally affected, the dominant task (word reading) showed priming effects on switch trials only. Thus, the retrieval of previous processing episodes has a selective impact on situations in which task competition is high. PMID- 16248326 TI - On the role of verbalization during task set selection: switching or serial order control? AB - Recent task-switching work in which paper-and-pencil administered single-task lists were compared with task-alternation lists has demonstrated large increases in task-switch costs with concurrent articulatory suppression (AS), implicating a crucial role for verbalization during switching (Baddeley, Chincotta, & Adlam, 2001; Emerson & Miyake, 2003). Experiment 1 replicated this result, using computerized assessment, albeit with much smaller effect sizes than in the original reports. In Experiment 2, AS interference was reduced when a sequential cue (spatial location) that indicated the current position in the sequence of task alternations was given. Finally, in Experiment 3, switch trials and no switch trials were compared within a block of alternating runs of two tasks. Again, AS interference was obtained mainly when the endogenous sequencing demand was high, and it was comparable for no-switch and switch trials. These results suggest that verbalization may be critical for endogenous maintenance and updating of a sequential plan, rather than exclusively for the actual switching process. PMID- 16248327 TI - Effects of response selection on the task repetition benefit in task switching. AB - A task switch typically leads to worse performance than a repetition does. This shift cost can be reduced with sufficient task preparation time, but a residual cost usually remains. We propose that a large part of this residual cost is caused by an activation bias produced by response selection processes in the preceding trial. In our experiments, we manipulated response selection requirements using a go/no-go methodology. The residual shift cost disappeared after no-go trials, suggesting that response selection is crucial to establish an activation bias for the current category-response rules and that this bias persists into the next trial. A comparison with a go-only group confirmed this analysis by revealing no differences in preparatory strategy due to the inclusion of no-go trials. In addition, no-go trials had no significant effects on subsequent trials in a single-task experiment, suggesting that no-go trials are not coded as a task different from go trials and that there is no inhibition of the prepared task in a no-go trial. We thus conclude that a persisting activation bias of response rules plays a major role in task switching. PMID- 16248328 TI - Central interference in error processing. AB - This study dealt with capacity limitations in error processing. Participants classified digits into three arbitrary categories (initial response). Half were required to correct their errors if an error was detected (correction response), and half were required to produce a second response, regardless of the correctness of the initial response (approval response). Auditory interference was introduced before, during, or after the initial response. Interference stimuli were to be recalled later and were, thus, considered to involve central processes. Results for before showed that although correction responses were elongated, approval responses given after erroneous initial responses were shortened. For during, both correction and approval responses were elongated. On the basis of our findings, we argue that the error process is generated before the erroneous response is given and that it is a central process in terms of being subjected to capacity limitations in the presence of other central processes. PMID- 16248329 TI - Attention control and ability level in a complex cognitive skill: attention shifting and second-language proficiency. AB - In this study, we investigated the relationship between attention control and proficiency in a complex cognitive skill. The participants were English-French bilinguals with varying degrees of second-language (French) proficiency. Proficiency was operationalized as efficiency of lexical access in an animacy judgment task, as reflected in the coefficient of variability of response time adjusted for first-language performance on the same task. Attention control was operationalized as the shift cost obtained in a linguistic version of the alternating runs task-switching paradigm. Hierarchical regression revealed that, overall, attention control accounted for 59% of the variance of proficiency and that second-language attention control alone accounted for 32% of the unique variance of proficiency, indicating a high degree of skill domain (second language) specificity in the relationship between attention control and proficiency. The results speak to issues regarding the development of expertise, second-language acquisition, and a cognitive linguistic approach to language and attention. PMID- 16248330 TI - Short-term memory and the attentional blink: capacity versus content. AB - When people monitor the rapid serial visual presentation (RSVP) of stimuli for two targets (T1 and T2), they often miss T2 if it falls into a time window of about half a second after T1 onset, a phenomenon known as the attentional blink (AB). We found that overall performance in an RSVP task was impaired by a concurrent short-term memory (STM) task and, furthermore, that this effect increased when STM load was higher and when its content was more task relevant. Loading visually defined stimuli and adding articulatory suppression further impaired performance on the RSVP task, but the size of the AB over time (i.e., T1 T2 lag) remained unaffected by load or content. This suggested that at least part of the performance in an RSVP task reflects interference between competing codes within STM, as interference models have held, whereas the AB proper reflects capacity limitations in the transfer to STM, as consolidation models have claimed. PMID- 16248332 TI - Depth cues do not underlie attentional modulations of the Stroop effect. AB - The well-known Stroop effect is usually attributed to the automaticity of word reading. Recently, Wuhr and Waszak (2003) had participants name the color of one of two rectangles and found that words in the relevant object produced larger Stroop effects than did words in the irrelevant object or in the background. They attributed this difference to an object-based mechanism of attentional selection that amplifies processing of all the features of an attended object. However, in the displays used by Wuhr and Waszak, occlusion suggested the presence of different depth planes. Hence, the increased Stroop effect could have resulted from perceiving the words to be in the same depth plane as the relevant object and not from perceiving the words to be parts of the relevant object. Two experiments tested between these accounts by using displays without monocular depth cues. The results of both experiments replicate those of Wuhr and Waszak, supporting their object-based account. PMID- 16248331 TI - Coherence of the irrelevant-sound effect: individual profiles of short-term memory and susceptibility to task-irrelevant materials. AB - We examined individual and developmental differences in the disruptive effects of irrelevant sounds on serial recall of printed lists. In Experiment 1, we examined adults (N = 205) receiving eight-item lists to be recalled. Although their susceptibility to disruption of recall by irrelevant sounds was only slightly related to memory span, regression analyses documented highly reliable individual differences in this susceptibility across speech and tone distractors, even with variance from span level removed. In Experiment 2, we examined adults (n = 64) and 8-year-old children (n = 63) receiving lists of a length equal to a predetermined span and one item shorter (span-1). We again found significant relationships between measures of span and susceptibility to irrelevant sounds, although in only two of the measures. We conclude that some of the cognitive processes helpful in performing a span task may not be beneficial in the presence of irrelevant sounds. PMID- 16248333 TI - Searching for the functional locus of the SNARC effect: evidence for a response related origin. AB - Dehaene, Bossini, and Giraux (1993) showed that when participants make parity judgments, responses to numerically small numbers are made faster with the left hand, whereas responses to large numbers are made faster with the right hand (the SNARC [spatial-numerical association of response codes] effect). According to one view, the SNARC effect arises at an early processing stage due to (in)congruencies between the digit's side of presentation and its representation on the mental number line, independently of response effector(s). Alternatively, the SNARC effect might arise at a later response-related stage due to (in)congruencies between the digit's representation on the mental number line and the side of response, independently of the side of presentation. The results of three experiments, using central and lateralized stimuli, and vocal and manual responses, clearly support the view that the SNARC effect arises at a relatively late response-related stage, without substantive contributions from earlier processing stages. PMID- 16248334 TI - Bodies and their parts. AB - How do we think about the space of bodies? Several accounts of mental representations of bodies were addressed in body part verification tasks. An imagery account predicts shorter times to larger parts (e.g., back < hand). A part distinctiveness account predicts shorter times to more discontinuous parts (e.g., arm < chest). Apart significance account predicts shorter times to parts that are perceptually distinct and functionally important (e.g., head < back). Because distinctiveness and significance are correlated, the latter two accounts are difficult to distinguish. Both name-body and body-body comparisons were investigated in four experiments. In all, larger parts were verified more slowly than smaller ones, eliminating the imagery/size account. Despite the correlation between distinctiveness and significance, the data suggest that when comparisons are perceptual (body-body), part distinctiveness is the best predictor, and when explicit or implicit naming is involved, part significance is the best predictor. Naming seems to activate the functional aspects of bodies. PMID- 16248335 TI - Spatial versus object visualizers: a new characterization of visual cognitive style. AB - The visual system processes object properties (such as shape and color) and spatial properties (such as location and spatial relations) in distinct systems, and neuropsychological evidence reveals that mental imagery respects this distinction. The findings reported in this article demonstrate that verbalizers typically perform at an intermediate level on imagery tasks, whereas visualizers can be divided into two groups. Specifically, scores on spatial and object imagery tasks, along with a visualizer-verbalizer cognitive style questionnaire, identified a group of visualizers who scored poorly on spatial imagery tasks but excelled on object imagery tasks. In contrast, a second group of visualizers scored high on spatial imagery tasks but poorly on object imagery tasks. The results also indicate that object visualizers encode and process images holistically, as a single perceptual unit, whereas spatial visualizers generate and process images analytically, part by part. In addition, we found that scientists and engineers excel in spatial imagery and prefer spatial strategies, whereas visual artists excel in object imagery and prefer object-based strategies. PMID- 16248336 TI - Gaze aversion: a response to cognitive or social difficulty? AB - When asked questions, adults and children often avert their gaze at certain points within the interaction, especially when questions are difficult (Doherty Sneddon, Bruce, Bonner, Longbotham, & Doyle, 2002; Glenberg, Schroeder, & Robertson, 1998). Gaze aversion may be a way of managing the cognitive load associated with the processing of visual environmental information, or it may serve to alleviate a negative social-emotional experience, such as the self consciousness associated with, for example, a fear of failure. In the present study, thirty-six 8-year-olds were questioned either face to face or across a live video link. Questions varied in type (arithmetic, verbal reasoning, and autobiographical and episodic memory) and in difficulty. Children averted their gaze more during face-to-face questioning than during video-mediated questioning; however, question difficulty had a very strong influence on aversion in both interview conditions. It is concluded that although social factors play a role in children's gaze aversion during pedagogical question-answer sequences, the primary function of averting gaze is to manage the cognitive load involved in the processing of environmental information. PMID- 16248337 TI - Chronometric evidence for memory retrieval in causal conditional reasoning: the case of the association strength effect. AB - According to many models of conditional reasoning, correct responses to the uncertain forms affirmation of consequent (AC) and denial of antecedent (DA) rely on the retrieval of an alternative antecedent from semantic memory. The main prediction issuing from this hypothesis of online retrieval is that the associative strength of the relevant information in long-term memory affects the latency of its retrieval and then of the correct response of uncertainty to AC and DA. This prediction was tested in 64 adults who performed a syllogism evaluation task with familiar causal conditional premises. As predicted, correct responses of uncertainty to AC and DA were not only more frequent but also faster when the antecedent term was weakly rather than strongly associated with the consequent. In line with the semantic framework, we also observed that this strength effect in response times was reversed for incorrect responses of certainty to AC. PMID- 16248338 TI - When logic fails: implicit transitive inference in humans. AB - Transitive inference (TI) in animals (e.g., choosing A over C on the basis of knowing that A is better than B and B is better than C) has been interpreted by some as reflecting a declarative logical inference process. We invert this anthropomorphic interpretation by providing evidence that humans can exhibit TI like behavior on the basis of simpler associative mechanisms that underlie many theories of animal learning. In this study, human participants were trained on a five-pair TI problem (A+B-, B+C-, C+D-, D+E-, E+F-) and, unlike in previous human TI studies, were prevented from becoming explicitly aware of the logical hierarchy, so they could not employ logical reasoning. They were then tested with three problems: B versus D, B versus E, and C versus E. Participants only reliably chose B over E, whereas the other test conditions yielded chance performance. This result is inconsistent with the use of logical reasoning and is instead consistent with an account developed to explain earlier TI studies with rats that found the same pattern of results. In this account, choice performance is based on differential associative strengths across the stimulus items that develop over training, despite equal overt reinforcement. PMID- 16248339 TI - A mixed-handed advantage in episodic memory: a possible role of interhemispheric interaction. AB - Recent behavioral and brain imaging data indicate that performance on explicit tests of episodic memory is associated with interaction between the left and right cerebral hemispheres, in contrast with the unihemispheric basis for implicit tests of memory. In the present work, individual differences in strength of personal handedness were used as markers for differences in hemispheric communication, with mixed-handers inferred to have increased interhemispheric interaction relative to strong right-handers. In Experiment 1, memory for words was assessed via recall or word fragment completion. In Experiment 2, memory for real-world events was assessed via recall. Results supported the hypothesis, in that mixed-handers displayed better episodic memory in comparison with strong right-handers. PMID- 16248340 TI - Garden variety. Project team's creativity blooms to eliminate institutional feel. PMID- 16248341 TI - Renovate or replace? Either way, a central utilities plant project is fraught with challenges. PMID- 16248342 TI - The new generation. Efficiencies, added services and high-end touches are priorities in today's rural hospital designs. PMID- 16248343 TI - Golden rules. Ten tips for avoiding common pitfalls in health care construction projects. PMID- 16248344 TI - Therapeutic environments. The increasingly documented connection between /design and care/. PMID- 16248345 TI - Speaking out on security. /NFPA/ develops new guidance document covering facility security issues. PMID- 16248346 TI - Bugs begone. Using /integrated pest management/ for environmentally responsible abatement. PMID- 16248347 TI - Bayesian designs with frequentist and Bayesian error rate considerations. AB - So far, most Phase II trials have been designed and analysed under a frequentist framework. Under this framework, a trial is designed so that the overall Type I and Type II errors of the trial are controlled at some desired levels. Recently, a number of articles have advocated the use of Bayesian designs in practice. Under a Bayesian framework, a trial is designed so that the trial stops when the posterior probability of treatment is within certain prespecified thresholds. In this article, we argue that trials under a Bayesian framework can also be designed to control frequentist error rates. We introduce a Bayesian version of Simon's well-known two-stage design to achieve this goal. We also consider two other errors, which are called Bayesian errors in this article because of their similarities to posterior probabilities. We show that our method can also control these Bayesian-type errors. We compare our method with other recent Bayesian designs in a numerical study and discuss implications of different designs on error rates. An example of a clinical trial for patients with nasopharyngeal carcinoma is used to illustrate differences of the different designs. PMID- 16248349 TI - Chance-corrected measures of reliability and validity in K x K tables. AB - When studying the degree of overall agreement between the nominal responses of two raters, it is customary to use the coefficient kappa. A more detailed analysis requires the evaluation of the degree of agreement category by category, and this is carried out in two different ways: using the value of kappa in the collapsed table for each category or using the agreement index for each category (proportion of agreements observed). Both indices have disadvantages: the former is sensitive to marginal totals; the latter is not chance corrected; and neither distinguishes the case where one of the two raters is a gold standard (an expert) from the case where neither rater is a gold standard. This article suggests five chance-corrected indices which are not sensitive to marginal totals and which differ depending on whether there is a standard rater. The article also justifies the reason for poor performance of kappa when the two marginal totals are unbalanced (especially if they are so in opposite directions) and the reason for its good performance when analysing the various 2 x 2 tables obtained by the collapse of a wider table. PMID- 16248348 TI - The role of proxy information in missing data analysis. AB - This article investigates the role of proxy data in dealing with the common problem of missing data in clinical trials using repeated measures designs. In an effort to avoid the missing data situation, some proxy information can be gathered. The question is how to treat proxy information, that is, is it always better to utilize proxy information when there are missing data? A model for repeated measures data with missing values is considered and a strategy for utilizing proxy information is developed. Then, simulations are used to compare the power of a test using proxy to simply utilizing all available data. It is concluded that using proxy information can be a useful alternative when such information is available. The implications for various clinical designs are also considered and a data collection strategy for efficiently estimating parameters is suggested. PMID- 16248351 TI - Outcome selection bias in meta-analysis. AB - Publication bias has been previously identified as a threat to the validity of a meta-analysis. Recently, new evidence has documented an additional threat to validity, the selective reporting of trial outcomes within published studies. Several diseases have several possible measures of outcome. Some articles might report only a selection of those outcomes, perhaps those with statistically significant results. In this article, we review this problem while addressing the questions: what is within-study selective reporting? how common is it? why is it done? how can it mislead? how can it be detected?, and finally, what is the solution? We recommend that both publication bias and selective reporting should be routinely investigated in systematic reviews. PMID- 16248350 TI - Assessing the reliability of ordered categorical scales using kappa-type statistics. AB - Methods for the analysis of reliability of ordered categorical scales are discussed, focussing on the limitation of the single summary-weighted kappa coefficients. A symmetric matrix of kappa-type coefficients is suggested as an alternative. The method is proposed as being suitable for ordinal scale where there is no underlying continuum. Their application is illustrated using two data sets from reliability studies. If, instead, distances between categories can be specified, a weighted mean of the matrix terms can be used as a summary measure. This is equal to a weighted kappa coefficient with squared weights, provided distances between adjacent categories are equal. When a study design corresponds to a one-way random effects model, estimates of precision of kappa-type coefficient, including the coefficients described here, can be obtained using the delta-method, bootstrap resampling by subjects or jack-knifing by subjects. In the case of interobserver reliability studies, where there may be systematic differences between observers, the investigator may wish to generalise to a population of observers and subjects. In this case, jack-knifing by observer and subject is suggested. Empirical comparisons are made between standard error estimates based on the delta-method, on jack-knifing by subjects and a two-way jack-knife by subjects and observers. The results suggest that standard errors based on the delta-method or jack-knifing by subject alone may be overly precise. PMID- 16248352 TI - [Leprosy situation in Myanmar before and after multidrug therapy]. AB - We introduced history of leprosy in Myanmar based on the book of Myanmar Academy of Medical Science published entitled "CONQUEST OF SCOURGES IN MYANMAR (Complied and Edited by Ko Ko, Kyaw and U Thaung) at 2002. "Leprosy Elimination Programme in Myanmar (Kyaw Lwin and Kyaw Nyunt Stein)" was appeared at chapter III in it. After dapsone treatment appeared, leprosy control program has started. Health system and service were developed and leprosy control program was also included in them. The integration of the elimination activities into basic health workers, such as midwives and health volunteers, has enabled the participation of a wide range of people in the community. After 1990s, multidrug therapy (MDT) was covered whole area of Myanmar, and task force for leprosy elimination was formed at Sate/Division, District and Township level. Finally Myanmar achieved the elimination of leprosy in January in 2003. PMID- 16248353 TI - [Immune dysregulation mediated by hepatitis C virus infection]. AB - Hepatitis C virus, which is non-cytopathic, establishes persistent infection in majority of patients after acute infection, causing various degrees of clinical liver disease. To escape and survive, hepatitis C virus may take ingenious strategies. Hepatitis C virus gene products interact host proteins to evade host immune responses in addition to the appearance of quasispecies. Against hepatitis C virus infection, host may avoid extensive tissue damage by inducing the activity of regulatory T cells. Insights into this mechanism of immune regulation may help to future development of novel therapies against hepatitis C virus. PMID- 16248354 TI - [JICA Leprosy Control and Basic Health Services Project in Myanmar]. AB - Japan International Cooperation Agency (JICA) implemented a 5-year long bilateral technical cooperation project, "Leprosy Control and Basic Health Services Project" in Myanmar. The project was implemented by National Leprosy Control Program, Department of Health with close technical collaboration of JICA experts mainly from International Medical Center of Japan (IMCJ) and National Sanatoriums of leprosy in Japan. It accelerated to achieve the elimination of leprosy at national level, which was declared in January 2003, and at sub-national level onward. It also developed the appropriate technologies for prevention of disability and prevention of worsening of disability (POD/POWD), which were introduced in 9 townships as a pilot service program. The Government stratified the POD/POWD services as a national program since 2005 by taking up the former pilot area to start with. The project also strengthened the function of referral system of leprosy control (Diagnosis and treatment), POD/POWD and physical rehabilitation. Beside leprosy, the project conducted a series of refresher trainings for primary health care givers, Basic Health Service Staff (BHS), of project areas (48 townships) to improve the services on tuberculosis, Malaria, Leprosy, Trachoma and HIV/AIDS for 3 years (2001-2003), which was evaluated in 2004. It contributed to improve the services at township level hospitals in procurement of audio-visual equipments and in conducting microscope training on leprosy, Malaria and tuberculosis at project areas. PMID- 16248355 TI - [Pathological diagnosis of leprosy in developing countries]. AB - In the developing countries where leprosy is prevalent, diagnosis of leprosy is made from clinical signs and symptoms. However, when difficult and doubtful cases increase after the advance of leprosy control programs, definitive diagnosis of leprosy by histopathology become necessary. This report describes our experience of technical support to re-establish histopathology service and introduction of immunohistochemistry in the leprosy referral center of Myanmar, and we discuss the ideal way of international technical support. This activity was performed as a part of leprosy control and basic health services project of Japan International Cooperation Agency (JICA) since 2000 to 2005. PMID- 16248356 TI - [Background of development and effectiveness of myanmar sandals (the activity of Leprosy Control and Basic Health Services Project in Myanmar)]. AB - "Leprosy control and Basic health service Project" was terminated at March 2005 within the scope of the period. I was assigned to develop the Plantar protect footwear for neuropathic feet, which called "Myanmar Sandals" in common. These sandals are not made by new technologies but made by basic knowledge. I would like to report about this activity as which is a part of distinguished strategy of ODA (Official Development Assistance) for developing country, but not as the technical report of these sandals. There is only one National Leprosy Hospital in Myanmar. At the time this project started, there were only two footwear technicians for the people who had disabilities by after effect of Hansen's disease in this country. And, it would appear that the number of the people is over 50,000. Furthermore, the budget of national hospital is not enough to refill necessary consumable supplies and materials. Although, I should instruct new technique which like making in Japan in ordinary circumstances, I could not think that such technical transfer by using costly imported materials woud bring beneficial effect and be continued. We were bound to be anxious about such situations when we made the plan. As a result of examining a plan based on these situations, we reached the conclusion that development of the simplified ready made footwear was pressing need. PMID- 16248357 TI - [Training of the rehabilitation staffs through taking measures against leprosy in Union of Myanmar]. AB - I participated in "Leprosy Control and Basic Health Service Project in Myanmar" by JICA from 2002 to 2004. I went to Myanmar three times as a short-term expert of physiotherapy. I carried out evaluation training for physio staffs in national Yenanthar leprosy hospital. They learned how to evaluate activity of daily living. And they were able to make modified spoons and the canes. Later, they looked for the contents to evaluate by themselves. And they evaluated and decided by themselves what they should do. They felt the necessity for toilet chairs, modified beds and etc. They formed the plan and manufactured them. They were able to perform by themselves. It is thought that evaluation was fixed to staffs. This knowledge will surely be helpful to patients. I expect that this knowledge will be put to evaluations and exercises of many diseases. The physio staffs of this hospital will surely perform. Cooperation of the shoemaker and nursing staffs is very secure. I hope that it may become the model of saying [working in cooperation with other occupational descriptions] from now on. PMID- 16248358 TI - [Survey on the present conditions of the residents regarding aging, aftereffects and subsequent complication]. AB - A survey of present conditions of the residents regarding aging, aftereffects and subsequent complication was conducted and its data were analyzed in the National Leprosarium Osima Seisho-en; located on an isolated island of the Seto Inland Sea. The results showed that they have become older with a various type of disease and most of them suffered from aftereffects caused leprosy, although after therapy. This survey also suggests that future problems in the management of our leprosarium will no doubt be to cope with the aging and to reducing our scale. PMID- 16248359 TI - [Effect of physician shortage on today's Hansen's disease management]. AB - In recent years, the total patient population of Hansen's disease has been rapid declining throughout Japan by as many as 200 - 250 each year. This decline is due to the aging of existing patients, and scarcity of new cases. Less patients are a welcome phenomenon. However, existing treatment facilities are now confronted with problems. The most serious problem is shortage of treating physicians. The causes of the shortage are (1) the aging of the existing patient population that creates mounting need for medical-surgical care and (2) the nation-wide shortage of physician in national sanatoria all over Japan. Let me present a sketch of the current situation at Amamiwakouen Hospital as an illustration. Amamiwakouen is a government-run sanatorium. It is responsible to take care of inpatient as well as out patients. Currently, there are 70 long term inpatients, who are handicapped by Hansen's disease sequela of various degrees. 11 minimally former inpatients are followed at the outpatient clinic. The clinic is also open to all community citizens. The number of annual clinic visitors averages 6,500 - 7,000. They come with wide range of problems requiring a dermatologist. While the total number of full-time staff is 100, only two are licensed physicians at present. This is indeed a disastrous situation. Therefore a number of contract physicians are recruited from nearby medical institutes to meat the demand. PMID- 16248360 TI - [Current endovascular therapy of cerebral aneurysm]. PMID- 16248361 TI - [Endovascular therapy for carotid artery stenosis]. PMID- 16248362 TI - [Up-to-date of current endovascular treatments for cerebral arteriovenous malformations]. PMID- 16248363 TI - [Dural arteriovenous malformation, dural arteriovenous fistula]. PMID- 16248364 TI - [Reversible posterior leukoencephalopathy syndrome (RPLS)]. PMID- 16248365 TI - [Cerebral circulation dynamics following fasudil intravenous infusion: a CT perfusion study]. AB - Using CT perfusion studies we evaluated changes in the cerebral circulation before and after the intravenous administration of fasudil 60 mg in 8 patients 7 to 14 days after a subarachnoid hemorrhage. The mean duration to the peak of the time-density curve and the average peak value did not change. In areas with cerebral blood perfusion (CBP) less than 40 ml/100 g/min, the CBP increased from 34.4 +/- 4.7 ml/100 g/min to 41.0 +/- 8.2 ml/100 g/min (p < 0.01) after fasudil infusion, the cerebral blood volume (CBV) rose from 2.41 +/- 0.53 ml/100 g to 2.55 +/- 0.5 ml/100 g (p < 0.05), and the mean transit time (MTT) decreased from 5.09 +/- 1.13 s to 4.82 +/- 0.89 s (p < 0.05). In areas where the CBP was more than 41 ml/100 g/min, the CBP did not change (from 51.8 +/- 7.6 ml/100 g/min to 50.4 +/- 8.4 ml/100 g/min), the CBV decreased (from 2.75 +/- 0.62 ml/100 g to 2.67 +/- 0.55 ml/100 g, p < 0.05), and the MTT did not change (from 3.80 +/- 0.76 s to 3.77 +/- 0.72 s). These results suggest that intravenous infusion of fasudil 60 mg increases cerebral blood flow and cerebral blood volume and shortens MTT in areas with decreased blood flow due to vasospasm. PMID- 16248366 TI - [A case report of acute neuromyopathy induced by concomitant use of colchicine and bezafibrate]. AB - We report a patient of acute neuromyopathy induced by concomitant use of colchicine and bezafibrate. A 75-year-old man with chronic renal failure and hyperlipidemia treated with bezafibrate (400 mg/day) for 1.5 years had developed watery diarrhea followed by acute tetraparesis, 14 days after the administration of colchicine for recurrent gout. Neurological examination showed proximal muscle weakness with myalgia, distal mild numbness (dysesthesia) of four limbs and generalized decreased or absent reflexies. The findings including elevated serum muscle enzymes, myogenic patterns with widespread myotonic discharge on the electromyography and delayed latency of F-wave on nerve conduction study indicated that the patient's clinical features were consistent with neuromyopathy. Soon after both colchicine and bezafibrate were stopped, the patient's symptoms resolved rapidly, therefore we made a diagnosis of drug induced neuromyopathy, although rhabdomyolysis with Guillain-Barre syndrome was initially suspected. Recently, there had been reported acute and severe neuromuscular disorder induced by combination therapy with colchicine and anti hyperlipidemic drugs, and there were clinical similarities between the cases of these reports and our case. Co-administration of colchicine with bezafibrate might accelerate the onset of neuromyopathy in connection with chronic renal failure in this case. Extreme caution is warranted when the patient with renal insufficiency concomitant use of colchicine and bezafibrate. PMID- 16248367 TI - [A case of spontaneous dissection of the cervical carotid artery with pseudoaneurysm]. AB - We experienced the case of a dissecting carotid artery presenting with a pulsatile neck mass. This 60-year-old man was admitted to our department due to a progressive, painful left neck mass. The lesion was diagnosed as a dissecting left internal carotid artery with the subsequent formation of a giant pseudoaneurysm. First, the patient underwent an endovascular treatment using self expandable stent and Guglielmi detachable coils (GDCs). However, four months after the treatment, recurrence of the pseudoaneurysm happened to him, and he was readmitted to our department. Angiographies revealed an enlargement of the pseudoaneurysm. Then, proximal occlusion of the left internal carotid artery was performed using a balloon and GDCs. Ten months after the second endovascular treatment, since symptoms of the central retinal artery embolism and progression of the mass effect occurred, surgical treatment with trapping and resection of the pseudoaneurysm was performed. We investigated pathologically the surgical specimen and could observe a partial thrombosed wall of the pseudoaneurysm. The multiple neovascular channels in the intima and media layers of the aneurysmal wall could be found, and the vasa vasorum in the adventitia was also noted. We thus suggest that mechanism of the enlargement of the pseudoaneurysm may be due to the retrograde neovascular supplies from the vasa vasorum. As a consequence, repeated bleeding and thrombosis in the dissecting arterial wall may result in the formation of partial thrombosed giant aneurysms. PMID- 16248368 TI - [Speech disturbance following myelography with iohexol: a case report]. AB - Iohexol (Omnipaque) is a non-ionic contrast media for neuroradiology, which causes the neurological complications on rare occasions. A 63-year-old, righthanded female suffered from dysphemia 12 hours after myelography with iohexol via lumbar puncture. The initial cerebral computed tomography scan revealed the generalized contrast media uptake into the cisterns, which was typical to the finding after intrathecal iohexol injection. Although the cerebral magnetic resonance (MR) imaging 7 days after attack also demonstrated no abnormal finding, the electrical encephalogram revealed the sporadic sharp wave activity. Her nonfluent speech deficit had recovered gradually, however, the neurogenic stuttering was still remained. Some literatures previously reported the cases with speech deficits as complications of metrizamide myelography, whereas, a case manifesting as speech disturbance following myelography with iohexol has been not reported. Metrizamide is also a non-ionic agent and had a lower incidence of speech disturbance, which is supposed to be associated with a focal superficial neurotoxic reaction of the cerebrum. Therefore, our unfortunate case suggests that iohexol could rarely cause speech disturbance such as metrizamide. PMID- 16248370 TI - [A case report of the diffuse axonal injury with special regards to the acute phase MRI findings]. PMID- 16248369 TI - [Fulminating midbrain irradiation injury of pediatric brain tumor]. AB - We report two children with post radiation midbrain damage causing severe neurological symptoms. A twelve-year-old boy with a four year history of hydrocephalus was diagnosed with tectal glioma, which endoscopic biopsy revealed to be low grade. He underwent gamma knife radiation surgery (central 24 Gy/peripheral 12 Gy). Two months later bilateral ptosis followed by total oculomotor palsy and drowsiness developed. Despite pulsed-steroid therapy the tumor size increased up to 4.6 times in volume. The tumor was totally removed and was diagnosed as an early delayed radiation reaction pathologically. His symptoms disappeared except for a slight upper gaze palsy. The second patient was a six year-old girl with a medulloblastoma. Following total resection and a VP shunt she received conventional radiation therapy along with chemotherapy. After the final irradiation she became comatose (JCS II-2) and MRI revealed diffuse midbrain damage with acute aqueduct obstruction, which recovered in two weeks. Reports of irradiation injuries of the midbrain in childhood are rare but it should be considered as a possible cause of fulminant symptoms requiring emergency treatment. Because of midbrain anatomical complexity, midbrain radiation therapy requires great care, especially in children. PMID- 16248371 TI - [A 59-year-old woman with personality change and abnormal behavior followed by amyotrophy and dementia]. AB - We report a 59-year-old woman with generalized amyotrophy and dementia. She showed personality change at 53 years of age. When she was 56 years old, she began to show abnormal and violent behaviors. At age 58, she developed dysphagia and amyotrophy of upper limbs. She was admitted to a hospital for the treatment of aspiration pneumonia. She was severely demented and showed pseudobulbar palsy, amyotrophy of tongues, weakness of upper limbs, and pyramidal signs. She was still able to walk by herself. Dementia, pseudobulbar palsy, and amyotrophy progressed rapidly. At age 59, she became bed ridden and required tube feeding. She died by aspiration pneumonia at age 59. The patient was discussed at a neurological CPC and the chief discussant arrived at the conclusion that the patient had ALS dementia. Other possibility discussed was Pick's disease with amyotrophy. Post-mortem examination revealed severe lower motor neuron degeneration. The upper motor neurons were unaffected. Neuronal loss was not observed in the cerebral cortex, but moderate gliosis was seen in the cerebral white matter. In addition, the substantia nigra was moderately degenerated. There were ubiquitin positive neuronal inclusions in the granular cells of the dentate gyrus. Also, Bunina bodies were seen in the neurons of spinal anterior horns. These findings were characteristic pathology for ALS with dementia. PMID- 16248373 TI - [Study of real-time PCR assays for rapid detection of food-borne pathogens]. AB - A duplex real-time SYBR Green LightCycler PCR (LC-PCR) assay with DNA extraction using QIAamp DNA Stool Minikit was evaluated for detection of 8 of 19 species of food-borne pathogens, including Plesiomonas shigelloides, Providencia alcalifaciens, in five stool specimens. The time frame was within 2h or less. The protocol used the same LC-PCR with 22 pairs of specific primers. The rapid amplification and reliable detection of specific genes were determined by this LC PCR assay from 10 cases of food-borne outbreaks in Shimane Prefecture from 2002 to 2005. These cases included Campylobacter jejuni (4), Clostridium perfringens (2), enteropathogenic Escherichia coli and astA positive E. coli (1), and astA positive E. coli, enterohemorrhagic E. coli 026, and Bacillus cereus (1 each). Rapid amplification and reliable detection of specific genes of food-or water borne pathogenic bacteria in fecal samples should facilitate the diagnosis and management of food-borne outbreaks. PMID- 16248372 TI - [Clinical effects of piperacillin and tazobactam/piperacillin on Haemophilus influenzae lower respiratory tract infection in pediatric patients]. AB - OBJECTIVE: The prevalence of beta-lactamase-nonproducing ampicillin-resistant (BLNAR) Haemophilus influenzae (H. influenzae) has been increasing in recent years. Piperacillin (PIPC) is one of a few beta-lactams possessing good activity against BLNAR H. influenzae. We studied clinical efficacy of piperacillin and its beta-lactamase inhibitor, tazobactam/piperacillin (TAZ/PIPC) in children with lower respiratory tract infection caused by H. influenzae including resistance strains. METHODS: Subjects were 20 children with lower respiratory tract infection caused by H. influenzae treated with PIPC 100mg/kg/day (7 cases) or TAZ/PIPC 125mg/kg/day (13 cases). We selected cases from which resistant H. influenzae strains might be detected. Patients received prior antimicrobial therapy within two weeks before admission, or with underlying diseases. We examined patient profiles, clinical efficacy, susceptibilities for 6 beta-lactam antibiotics [PIPC, TAZ/PIPC, ampicillin (ABPC), cefotaxime (CTX), ceftriaxone (CTRX), and meropenem (MEPM)] and analyzed 6 genotype patterns of beta-lactam resistant genes by PCR. RESULTS: Efficacy was 7/7 in patients in PIPC group and 12/13 in patients in TAZ/PIPC group. Diminished efficacy was seen in only one case complicated with severe RSV infection. The susceptibility of all strains but one beta-lactamase producing, ABPC resistant (BLP) strain to PIPC and of all to TAZ/ PIPC was below 0.25 microg/mL. The genotype of the 15 strains isolated from the sputum on administration was as follows; beta-lactamase nonproducing, ABPC susceptible (gBLNAS) strains were 4, gBLP strain was 1, beta-lactamase nonproducing, and ABPC-resistant (gLow-BLNAR) strains were 2, beta-lactamase nonproducing, ABPC resistant (gBLNAR) strains were 8. CONCLUSION: PIPC and TAZ/PIPC were useful against lower respiratory tract infection caused by H. influenzae including BLNAR in children. PMID- 16248374 TI - [Clinical studies of sixteen cases with pulmonary cryptococcosis mainly with respect to serum level of cryptococcal antigen]. AB - Clinical studies of sixteen cases with pulmonary cryptococcosis, during the past six years between 1998 and 2004, were peformed mainly with respect to serum cryptococcal antigen titer. Serum cryptococcal antigen was positive in twelve of 16 cases, the other three cases were diagnosed by VATS, the other one by positive culture of cryptococcus in BALF. In these twelve cases, the serum cryptococcal antigen titer was continuously tested after treatment. The serum cryptococcal antigen titer decreased from half to 6 months after treatment. And the cryptococcal Ag changed to negative in six of the 12 cases by antifungal agents from 5 to 19 months. But four cases whose pneumonia was severe tended to have a high titer level of cryptococcal antigen and were positive for a long period. In the Chest CT of four pulmonary cryptococcosis case with negative cryptococcal antigen, all of the maximum nodule size was less than or equal to 15mm in diameter. PMID- 16248376 TI - [Cluster analysis of restriction fragment length polymorphism patterns of Mycobacterium tuberculosis isolated in Chiba prefecture]. AB - Methods for cluster analysis of IS6110 based restriction fragment length polymorphism (RFLP) patterns of Mycobacterium tuberculosis isolates were studied for an epidemiological investigation in Chiba prefecture. To normalize patterns, external size markers were adopted instead of typical internal size markers used in the standard method. RFLP patterns were run on 1.4% agarose gels and external markers were applied to outside and middle lanes on each gel for precise comparison. The resulting RFLP patterns of 74 isolates were clustered by similarity. Similarity was calculated with the Dice coefficient using parameter settings at 0.8% tolerance and 0.5% optimization. Patterns of 19 isolates from 8 outbreaks showed high similarity within each outbreak. Cluster analysis, as described here, provides insights into epidemiological tracing of tuberculosis in Chiba prefecture. PMID- 16248375 TI - [Relationship between ABO histo-blood group type and an outbreak of norovirus gastroenteritis among primary and junior high school students: results of questionnaire-based study]. AB - Noroviruses are common causative agents of epidemic gastroenteritis in humans. Recent studies showed that human susceptibility to noroviruses was associated with ABO histo-blood group type. It was also observed that various degrees of susceptibility were exhibited by different norovirus strains. In January 2003, an outbreak of acute gastroenteritis including 661 affected primary and junior high school students occurred through lunch bread contaminated with norovirus in Hokkaido, Japan. To clarify the relationship between ABO histo-blood group type and the norovirus infection, we performed a written questionnaire to schoolchildren about the consumption of the bread, onset of symptoms and person to-person transmission in their household. Questionnaires were returned from 722 schoolchildren (response rate, 65.8%), of whom 55.3% suffered gastroenteritis. As a result of this survey, it was found that schoolchildren with blood group type A (71.1%, 133/187) were more susceptible to the norovirus infection, whereas, schoolchildren with blood group type AB (55.3%, 26/47) were less affected (P (Z0) < 0.025). In addition, the presumptive prevalence rate of person-to-person transmission in each household indicated that schoolchildren with blood group type AB (19.2%, 5/26) had a lower risk of infection than those with blood group type A or O [A : 41.4%, 55/133 O : 39.5%, 49/124 (unknown for one case) ] [P (Z0) < 0.025]. Our findings suggested that persons with blood group type AB were less affected by the norovirus infection in this outbreak. PMID- 16248377 TI - [A study of bacterial meningitis in Hokkaido between 1999 and 2003]. AB - We evaluated 83 children with bacterial meningitis in 67 of 81 insitutions in Hokkaido, Japan between 1999 and 2003 by questionnaire. The incidence of bacterial meningitis in children aged < 5 years and 5 < or =, < 10 years was 6.3 and 0.7 cases per 100, 000 children/year. The incidences of meningitis due to Haemophilus influenzae, Streptococcus pneumoniae, and Streptococcus agalactiae in children aged < 5 years were 3.7, 1.4 and 0.8. Median age was 13 months ranging from 0 day to 9 years. Seventy-four (89.1%) of the total were less than 5 years old with 39 (47.0%) less than 1 year old. Major causative organisms were H. influenzae in 51 patients, S. pneumoniae in 18, S. agalactiae in 9 and E. coli in 3. Four deaths occurred, giving an overall case mortality rate of 4.8%. Sequelae were seen at discharge in 25.3%, predominantly epilepsy, hearing loss, and developmental delay of varying severity. The frequency of poor prognosis was 55.6% for patients with meningitis due to S. pneumoniae compared with 21.6% for H. influenzae and 22.2% for S. agalactiae. PMID- 16248378 TI - Use of reference tissue models for quantification of histamine H1 receptors in human brain by using positron emission tomography and [11c]doxepin. AB - The aim of the present study is to evaluate the validity of the simplified reference tissue model (SRTM) and of Logan graphical analysis with reference tissue (LGAR) for quantification of histamine H1 receptors (H1Rs) by using positron emission tomography (PET) with [11C]doxepin. These model-based analytic methods (SRTM and LGAR) are compared to Logan graphical analysis (LGA) and to the one-tissue model (1TM), using complete datasets obtained from 5 healthy volunteers. Since HIR concentration in the cerebellum can be regarded as negligibly small, the cerebellum was selected as the reference tissue in the present study. The comparison of binding potential (BP) values estimated by LGAR and 1TM showed good agreement; on the other hand, SRTM turned out to be unstable concerning parameter estimation in several regions of the brain. By including the results of noise analysis, LGAR became a reliable method for parameter estimation of [11C]doxepin data in the cortical regions. PMID- 16248379 TI - Assessment of Takotsubo cardiomyopathy (transient left ventricular apical ballooning) using 99mTc-tetrofosmin, 123I-BMIPP, 123I-MIBG and 99mTc-PYP myocardial SPECT. AB - We compared Takotsubo cardiomyopathy (transient left ventricular apical ballooning) with acute myocardial infarction (AMI) using two-dimensional echocardiography, 99mTc-tetrofosmin, 99mTc-PYP, 123I-BMIPP and 123I-MIBG myocardial SPECT. METHODS: We examined 7 patients with Takotsubo cardiomyopathy and 7 with AMI at the time of emergency admission (acute phase), and 2-14 days (subacute phase), one month (chronic phase), and 3 months (chronic II phase) after the attack. The left ventricle was divided into nine regions on echocardiograms and SPECT images, and the degree of abnormalities in each region was scored according to five grades from normal (0) to severely abnormal (4). RESULTS: Coronary angiography showed the absence of stenotic regions in patients with Takotsubo cardiomyopathy, and severely stenotic and/or occlusive lesions in patients with AMI. The total ST segment elevation on electrocardiograms (mm) was 7.8 +/- 3.7 in those with Takotsubo cardiomyopathy, and 7.3 +/- 3.9 in patients with AMI. Abnormal wall motion scores on echocardiograms were 14.2 +/- 4.6, 4.7 +/- 4.0, 1.7 +/- 2.0 and 0.5 +/- 0.4 during the acute, subacute, chronic and chronic II phases, respectively, in patients with Takotsubo cardiomyopathy, and 14.0 +/- 4.3, 11.4 +/- 3.9, 8.8 +/- 3.6 and 5.2 +/- 4.8 in those with AMI. Abnormal myocardial perfusion scores on 99mTc-tetrofosmin images were 11.8 +/- 3.5, 3.2 +/- 3.0, 0.5 +/- 1.2 and 0.2 +/- 0.4 during the acute, subacute, chronic and chronic II phases, in patients with Takotsubo cardiomyopathy, and 16.2 +/- 4.3, 13.9 +/- 4.6, 7.9 +/- 4.6 and 5.0 +/- 4.5, respectively, in those with AMI. Abnormal myocardial fatty acid scores on 123I-BMIPP images were 12.6 +/- 3.7, 6.8 +/- 3.2 and 0.4 +/- 0.6 during the subacute, chronic and chronic II phases, respectively, in patients with Takotsubo cardiomyopathy, and 16.5 +/- 5.1, 14.7 +/- 4.8 and 7.5 +/- 4.5 in those with AMI. Abnormal myocardial sympathetic nerve function scores on 123I-MIBG images were 14.8 +/- 4.0, 8.8 +/- 4.0 and 0.4 +/- 0.6 during the subacute, chronic, chronic II phases, respectively, in patients with Takotsubo cardiomyopathy, and 18.6 +/- 6.5, 16.8 +/- 6.8 and 12.9 +/- 5.2 in those with AMI. Myocardial 99mTc-PYP uptake was abnormal not only in patients with AMI but also in those with Takotsubo cardiomyopathy during the acute phase. CONCLUSIONS: Takotsubo cardiomyopathy might represent a stunned myocardium caused by a disturbance of the coronary microcirculation. PMID- 16248380 TI - Left ventricular systolic/diastolic function evaluated by quantitative ECG-gated SPECT: comparison with echocardiography and plasma BNP analysis. AB - OBJECTIVE: The aim of this study was to evaluate the left ventricular (LV) functional parameters calculated using quantitative electrocardiography (ECG) gated myocardial perfusion single photon emission computed tomography (QGS). In addition to LV systolic parameters, diastolic parameters were compared with those by ultrasound echocardiography (UCG) and also with plasma B-type natriuretic peptide (BNP) concentrations. METHODS: We examined 46 patients with various forms of heart disease. By the QGS data with 16 framing data acquisition using technetium (Tc)-99m methoxyisobutylisonitrile (MIBI) perfusion, we calculated the following parameters: LV end-diastolic volume (EDV), end-systolic volume (ESV), ejection fraction (EF), peak filling rate (PFR), filling rate during the first third of the filling time (1/3FR) and first third filling fraction (1/3FF). By UCG, we measured mitral early to atrial (E/A) wave velocity ratio and pulmonary venous inflow systolic/diastolic (S/D) ratio as diastolic functional parameters. Plasma BNP concentrations were also measured. RESULTS: There was a significant correlation between LVEDV, ESV and EF measured by QGS and UCG (EDV, r = 0.71, p < 0.001; ESV, r = 0.82, p < 0.001; EF, r = 0.75, p < 0.001). The PFR, 1/3FR and 1/3FF obtained by QGS correlated positively with E/A ratio (PFR, r = 0.54, p < 0.001; 1/3FR, r = 0.61, p < 0.001; 1/3FF, r = 0.42, p < 0.01) and negatively with S/D ratio (PFR, r = -0.40, p < 0.01; 1/3FR, r = -0.38, p < 0.05; 1/3FF, r = 0.39, p < 0.01) obtained by UCG. Plasma BNP concentrations in EF < 50% patients were greater than those in EF > or = 50% patients (335.2 +/- 60.2 vs. 101.2 +/- 41.3 pg/ml, p < 0.01, both n = 17). Plasma BNP levels were also compared between higher and lower 1/3FF patients matched for LVEF. Plasma BNP concentrations in 1/3FF < 35% patients were significantly greater than those in 1/3FF > or = 35% patients (312.9 +/- 62.5 vs. 120.5 +/- 32.8 pg/ml, p < 0.05, both n = 14). CONCLUSIONS: The degree of LV systolic and diastolic dysfunctions evaluated by QGS correlated with that by UCG or BNP. The QGS functional parameters offer useful information regarding cardiac failure. PMID- 16248381 TI - Detection of alveolar epithelial injury by Tc-99m DTPA radioaerosol inhalation lung scan in rheumatoid arthritis patients. AB - Rheumatoid arthritis (RA) is a systemic autoimmune disorder primarily involving the joints. Lung alterations in RA may be primary or secondary to pharmacological treatments and may involve the alveoli, interstitium, airways and/or pleura. Technetium-99m diethylenetriaminepentaacetic acid (Tc-99m DTPA) aerosol inhalation scintigraphy is a sensitive and noninvasive test commonly employed to assess pulmonary epithelial membrane permeability. The purpose of the this study was a) to investigate the changes of pulmonary alveolar epithelial permeability in patients with RA, b) to determine the relationship between the clearance rate of Tc-99m DTPA and pulmonary function test (PFT) results, and c) to determine the relationship between the clearance rete of Tc-99m DTPA and clinical parameters of disease. Twenty-five patients with RA but without lung alterations were included in the study. The patients were 22 females, and 3 males; mean age 53.6 +/- 8.7 years. Technetium-99m DTPA aerosol inhalation scintigraphy was performed on the study and healthy control groups. Clearance half times (T1/2) were calculated by placing a mono-exponential fit on the curves. Penetration index (PI) was calculated on the first-minute image. There were no significant differences in the mean T1/2 or mean PI values between the RA patients and control subjects. No correlation was found between the mean T1/2 values of Tc-99m DTPA clearance and activity of RA, clinical values, or the spirometric measurements except FEV1/FVC and functional status in RA patients (p = 0.02, p = 0.01, respectively). However, a weak correlation was found between duration of disease and T1/2 values of Tc 99m DTPA clearance (p = 0.006). PI values tended to correlate with FEF25-75, although, this was not statistically significant (p = 0.057). This study shows that no changes occur in alveolar-capillary permeability in RA patients without lung alterations. PMID- 16248382 TI - Thallium-201 scintigraphy in bone and soft-tissue tumors: a comparison of dynamic, early and delayed scans. AB - OBJECTIVE: It has been reported that delayed scan of thallium-201 (201Tl) scintigraphy is useful for differentiating malignant tumors from benign lesions and for evaluating treatment response. However, physiological muscle uptake which usually increases in delayed scans, often makes it difficult to evaluate 201Tl uptake and its washout in bone and soft-tissue tumors. The purpose of this study was to evaluate whether the delayed scan is necessary and whether a dynamic scan is useful in the evaluation of bone and soft-tissue tumors. METHODS: We studied 175 cases of bone and soft-tissue tumors (malignant 45, benign 130). Dynamic scans were acquired every 5 seconds for 10 minutes after 201Tl injection, and time activity curves (TACs) were generated by adaptive smoothing methods. Early and delayed scans were acquired at 10-15 minutes and 2 hours after injection. 201Tl images were visually interpreted and the radioactivity count ratio (T/N) of tumors to normal tissues and washout rate [WR = (early T/N - delayed T/N)/early T/N] were defined. RESULTS: When there were no 201Tl uptake in dynamic (n = 67) and early scans (n = 68), no tumor uptake was also appreciated in delayed scans, and all but two cases of negative scans were benign. In 107 lesions, although there were significant differences in T/Ns between malignant and benign lesions both on early scans (2.84 +/- 1.45 vs. 2.05 +/- 1.13, p < 0.05) and delayed scans (2.17 +/- 1.03 vs. 1.58 +/- 0.64, p < 0.05), there was a substantial overlap. The T/Ns decreased in delayed scans (i.e., WR > 0) in 100 of 107 cases due to increase of surrounding muscle uptake, and there was no difference in WR between malignant tumors and benign lesions (0.21 +/- 0.14 vs. 0.19 +/- 0.14). CONCLUSIONS: For evaluating bone and soft-tissue tumors, delayed scan had little clinical usefulness and it may be time consuming. Dynamic scan would be useful for demonstrating the differences between tumor blood flow and 201Tl uptake in tumors. PMID- 16248383 TI - Usefulness of noise adaptive non-linear gaussian filter in FDG-PET study. AB - OBJECTIVE: In positron emission tomography (PET) studies, shortening transmission (TR) scan time can improve patient comfort and increase scanner throughput. However, PET images from short TR scans may be degraded due to the statistical noise included in the TR image. The purpose of this study was to apply non-linear Gaussian (NLG) and noise adaptive NLG (ANLG) filters to TR images, and to evaluate the extent of noise reduction by the ANLG filter in comparison with that by the NLG filter using phantom and clinical studies. METHODS: In phantom studies, pool phantoms of various diameters and injected doses of 2-deoxy-2 [18F]fluoro-D-glucose (FDG) were used and the coefficients of variation (CVs) of the counts in the TR images processed with the NLG and ANLG filters were compared. In clinical studies, two normal volunteers and 13 patients with tumors were studied. In volunteer studies, the CV values in the liver were compared. In patient studies, the standardized uptake values (SUVs) of tumors in the emission images were obtained after processing the TR images using the NLG and ANLG filters. RESULTS: In phantom studies, the CV values in the TR images processed with the ANLG filter were smaller than those in the images processed with the NLG filter. When using the ANLG filter, their dependency on the phantom size, injected dose of FDG and TR scan time was smaller than when using the NLG filter. In volunteer studies, the CV values in the images processed with the ANLG filter were smaller than those in the images processed with the NLG filter, and were almost constant regardless of the TR scan time. In patient studies, there was an excellent correlation between the SUVs obtained from the images with a TR scan time of 7 min processed with the NLG filter (x) and those obtained from the images with a TR scan time of 4 min processed with the ANLG filter (y) (r = 0.995, y = 1.034x - 0.075). CONCLUSIONS: Our results suggest that the ANLG filter is effective and useful for noise reduction in TR images and shortening TR scan time while maintaining the quantitative accuracy of FDG-PET studies. PMID- 16248384 TI - The clinical value of dacryoscintigraphy in the selection of surgical approach for patients with functional lacrimal duct obstruction. AB - PURPOSE: Dacryoscintigraphy is widely known to be an effective modality in diagnosing abnormalities of the lacrimal system that cause epiphora (pathological overflow of tear). However, dacryoscintigraphy rarely serves beyond the simple diagnostic use for lacrimal duct obstruction. In our study, dacryoscintigraphy results of patients with functional lacrimal duct obstruction are newly classified into three types, the effects and prognoses of silicone tube intubation are noted according to each type, and the role of dacryoscintigraphy in determining appropriate surgical approaches is evaluated. METHODS: Subjects were 36 eyes of 29 patients complaining of epiphora who had increased tear meniscus, but showed no sign of obstruction on duct syringing. Impression of functional lacrimal duct obstruction was made through dacryoscintigraphy, and silicone tubes were inserted. RESULTS: Patients were classified according to the results of dacryoscintigraphy; those with delayed secretion in the distal nasolacrimal duct were typed as class I; those with delays in the proximal nasolacrimal duct class II; and delayed secretion from the pre-lacrimal sac to the lacrimal sac as class III. All patients had silicone tube intubations together with selective punctoplasty. Symptomatic improvement was observed in all 6 cases of distal nasolacrimal duct obstruction (100%), 14 of 18 proximal obstruction cases (77.8%), and 8 of 12 pre-lacrimal obstructions (66.7%). CONCLUSIONS: Functional lacrimal duct obstruction is easily diagnosed with dacryoscintigraphy. Furthermore, its may be classified by types of obstruction to predict post-operative results of silicone tube insertion. Cases suspicious of pre-lacrimal sac obstructions in particular may achieve better operative results with adjuvant treatments in addition to silicone tube insertion. PMID- 16248386 TI - Contribution of whole body FDG-PET to the detection of distant metastasis in pancreatic cancer. AB - OBJECTIVE: Accurate baseline staging is necessary to appropriately treat pancreatic cancer. The present study was undertaken to evaluate the clinical contribution of whole body FDG-PET to the detection of distant metastasis in pancreatic cancer. METHODS: A total of consecutive 42 patients with previously untreated pancreatic cancer were examined. Whole body FDG-PET imaging for initial staging was performed with a 3D acquisition and iterative reconstruction on Siemens ECAT HR+ scanner at 1 hour post 185-200 MBq 18F-FDG injection. PET findings were correlated with clinical and radiological data to determine the impact of PET on staging. RESULTS: In 16 patients, there were one or more sites of metastasis based on clinical data. FDG-PET correctly identified the presence of metastasis in 13 of 16 patients and its absence in 23 of the remaining 26 patients. Thus, FDG-PET missed 4 metastatic sites in 4 patients (liver and lung metastasis). FDG-PET correctly identified 8 metastatic sites in 7 patients (peritoneal dissemination and liver, bone and supraclavicular lymph node metastasis), which were missed on CT imaging. Based on whole body FDG-PET, the clinical stage was changed in 5 of 42 patients (11.9%). CONCLUSIONS: These results suggest that FDG-PET and CT appear to have a complementary role in the detection of distant metastasis in patients with pancreatic cancer. PMID- 16248385 TI - Initial experience with X-ray CT based attenuation correction in myocardial perfusion SPECT imaging using a combined SPECT/CT system. AB - OBJECTIVE: Attenuation artifacts adversely affect the diagnostic accuracy of myocardial perfusion imaging. We assessed the clinical usefulness of X-ray CT based attenuation correction (AC) in patients undergoing myocardial perfusion imaging by comparing their myocardial AC- and non-corrected (NC) SPECT images with the coronary angiography (CAG). METHODS: We retrospectively reviewed the myocardial SPECT images of 30 patients (18 men, 12 women; mean age 68 years). Thirteen of 30 patients with coronary artery disease (CAD) and 17 without CAD were confirmed by CAG. They underwent sequential CT and myocardial SPECT imaging with thallium-201 (111 MBq) under an exercise or pharmacological stress protocol using our combined SPECT/ CT system. Two readers reviewed the myocardial SPECT images for the presence of CAD on a 4-point scale where 1 = normal, 2 = probably normal, 3 = probably abnormal, and 4 = abnormal. Two reading sessions were held. First, non-corrected (NC)-SPECT and second, AC-SPECT images using X-ray CT images were interpreted. Interobserver variability was assessed with kappa statistics. Diagnostic performance (accuracy) of coronary arterial stenosis was compared between AC- and NC-images. RESULTS: Interobserver agreement for visual assessment was substantial or almost perfect. For AC-images, the observer consensus for analysis was 0.84 for the LAD-, 0.87 for the LCX-, and 0.71 for the RCA territory. For NC-images, it was 0.91, 0.71, and 0.78. AC resulted in statistically significant improvements in overall diagnostic accuracy (sensitivity/ specificity/accuracy = 76%/93%/89%, 67%/86%/81%, respectively, for AC- and NC-images). CONCLUSIONS: Because of an increase in the specificity, diagnostic accuracy was significantly increased on AC-images. These preliminary data suggest that X-ray CT based AC in myocardial SPECT imaging has the potential to develop into a reliable clinical technique. PMID- 16248387 TI - Binding kinetics of 11C-N-methyl piperidyl benzilate (11C-NMPB) in a rhesus monkey brain using the cerebellum as a reference region. AB - The binding kinetics of 11C-N-methyl piperidyl benzilate (11C-NMPB) in rhesus monkey brain were studied using animal positron emission tomography (PET) (SHR2000). This study is intended to assess the validity of the method using the cerebellum as a reference region, and to evaluate the effects of anesthesia on 11C-NMPB binding. Two monkeys, anesthetized with ketamine, received intravenous 11C-NMPB alone (370-760 MBq, < 1 microg/kg) or mixed with varying doses of nonradioactive NMPB (3 microg/kg, 10 microg/kg, 30 microg/kg) and were subjected to PET scans for 60 minutes. Regions of interest (ROI) were drawn on reconstructed PET images and a time-activity curve was obtained for each region. 11C-NMPB accumulated densely in the striatum and cerebral cortex with time. In contrast, the tracer accumulation significantly decreased with increased doses of nonradioactive NMPB. In the cerebellum, on the other hand, the accumulation of 11C-NMPB remained low and the tracer was slowly eliminated from the brain following the injection. 11C-NMPB binding in the cerebellum was barely affected by the increased dose of nonradioactive NMPB. We thus concluded that the specific 11C-NMPB binding was negligible in the cerebellum, and performed simplified evaluation of 11C-NMPB binding in each brain region by a graphical method using the cerebellum as a reference region. PET was conducted 26 times, in total both in ketamine-anesthetized and awake monkeys (n = 3 each). Measurements of 11C-NMPB binding showed good run-to-run reproducibility within individual animals. When 11C-NMPB binding was compared between ketamine-treated and awake animals, a significant increase in 11C-NMPB binding was observed in the striatum but not in other brain regions of ketamine-treated animals. PMID- 16248388 TI - Unfused renal ectopia: a rare form of congenital renal anomaly. AB - Unfused crossed renal ectopia observed 1 in 75,000 autopsies is a rare congenital anomaly. Typically one kidney is located in the proximity of the other kidney, and the ureter of the anatomically anomalous kidney crosses the midline to insert to the bladder in its normal anatomic position. Although renal function is usually not affected, the condition is generally accompanied by other congenital anomalies. In this case report, static and dynamic scintigraphic images of two patients with unfused crossed renal ectopia are presented. Besides properties of imaging modalities, clinical features are discussed in light of the available literature. PMID- 16248389 TI - Unilateral temporary functional stasis in the upper urinary tract caused by "a filled bladder" on Tc-99m DTPA diuresis renography: a teaching case. AB - A 2-year-old girl with recurrent urinary tract infection having slight left pelvicaliceal dilatation on her renal ultrasound underwent a Tc-99m DTPA diuresis renography. During the excretion phase, a prominent and persisting left pelvicaliceal stasis was noticed even after the diuretic injection. However, it disappeared simultaneously with an uncontrolled micturition. This patient is presented to show the effect of filled bladder on the physiological drainage of urine. Since urine flow in the urinary system is more complicated than simple drainage, a thorough understanding of the physiological basis for diuresis renography and the pitfalls of the technique is required for its appropriate use in the management of patients suspected of urinary tract obstruction. PMID- 16248390 TI - Cold tuberculous abscess identified by FDG PET. AB - We report FDG PET of two cases of cold abscess due to Mycobacterium tuberculosis. Case 1 had colon cancer; FDG PET showed high FDG uptake in the colon lesion and low uptake in the inguinal lesion. The latter was a tuberculous cold abscess confirmed by CT/MRI and biopsy. Case 2 received radiotherapy for lung cancer and presented with suspected vertebral metastasis. Further studies revealed tuberculosis of the vertebra and a tuberculous cold abscess in the iliopsoas muscle. FDG PET showed moderate uptake in the third lumbar spine and low uptake in the abscess center of iliopsoas lesion. Both tuberculous cold abscesses showed moderate FDG uptake in the capsule and low uptake in the center. These features are unique compared with non-tuberculous abscess and typical tuberculosis lesions, which are characterized by high FDG uptake. Pathologically, tuberculous cold abscess is not accompanied by active inflammatory reaction. Our findings suggested that the FDG uptake by tuberculous lesion varies according to the grade of inflammatory activity. The new diagnostic features of tuberculous cold abscess may be useful in the evaluation of such lesions by FDG PET. PMID- 16248391 TI - A case of diffuse hepatic angiosarcoma diagnosed by FDG-PET. AB - A 76-year-old woman was admitted to our hospital with a 2-month history of increasing abdominal distension, leg edema, and dyspnea. The serum transaminase level was about twice the upper limit of normal. The CT showed no tumor. Fluorine 18 2-deoxy-2-fluoro-D-glucose (FDG)-positron emission tomography (PET) showed diffuse abnormal accumulation throughout the entire liver. She was diagnosed by histopathological examination as having hepatic angiosarcoma causing veno occlusive disease (VOD). This is the first report of hepatic angiosarcoma with FDG-PET. PMID- 16248392 TI - Clearance of technetium-99m-labeled DTPA in hyperthyroidism without clinical evidence of lung disease, and relation to pulmonary function. AB - OBJECTIVE: The mechanisms of dyspnea and exercise intolerance have not been fully elucidated. We aimed to investigate the clearance rate of technetium-99m diethyltriaminepentaaceticacid (Tc-99m DTPA) from lungs in hyperthyroid patients without clinical evidence of lung disease and to explore the interactions between their Tc-99m DTPA radioaerosol lung scintigraphy, spirometric measurements, and the levels of thyroid hormones. METHODS: We studied 19 hyperthyroid patients and 16 sex- and age-matched controls. Thyroid hormone levels were assessed. Spirometric lung function tests, diffusing capacity of the lung for carbon monoxide (DLCO) and the clearance rate of Tc-99m DTPA were performed in all participants. Ratio of DLCO value to the alveolar ventilation (DLCO/VA) and the means of half-time (T1/2) of Tc-99m DTPA clearance rate, which were used to evaluate alveolar-capillary membrane permeability, were calculated. RESULTS: There were no statistical differences between spirometric parameters (VC, FVC, FEV1/FVC, FEF 25-75) of the two groups (p > 0.05). Although the mean FEV1 level was significantly lower in the hyperthyroid patients than the control subjects (p < 0.01), in five patients FEV1 was only less than 80 percent of the predicted value. No significant difference in the means of DLCO, DLCO/VA or T1/2 values of Tc-99m DTPA clearance was observed between the two groups (p > 0.05). In hyperthyroid patients, there was a positive relation between DLCO/VA, DLCO/VA % and T1/2 values of Tc-99m DTPA clearance (p < 0.01, r = 0.732, p < 0.01, r = 0.742, respectively). The lung volumes and the levels of thyroid hormones did not show a significant relationship to T1/2 values of Tc-99m DTPA clearance in hyperthyroid group (p > 0.05). CONCLUSIONS: We conclude that increased thyroid hormones have no effect on permeability of alveolar-capillary membrane in hyperthyroid patients. PMID- 16248393 TI - [Long-term prognosis of analgesics abuse headache]. AB - OBJECTIVE: To evaluate the long-term prognosis of analgesics abuse headache. METHODS: Ninety-five consecutive patients with analgesics abuse headache were treated in Toyonaka Municipal Hospital. Seventy-three patients (76.8%) had migraine, eighteen (19.0%) had tension-type headache and four (4.2%) new daily persistent headache. Seventy-seven (81.1%) were females and eighteen (18.9%) males. All patients were treated for six years from November 1997 to October 2003 and a total of sixty-nine patients were available for interview as of October 2004 at a mean time interval of 41.5 months after drug withdrawal therapy. Twenty two patients were admitted to our inpatient withdrawal unit, twenty-five patients were treated by outpatient withdrawal therapy. Tapering analgesics gradually in conjunction with instituting preventive therapy treated twenty-two patients. RESULTS: Inpatient-Nine cases (41%) reported intake of analgesics on < or = 8 days/month, five cases (23%) on 9-15 days/month and eight cases (36%) > 15 days/month. Three cases (14%) developed recurrent analgesic abuse. Outpatient (abrupt discontinuation)--Twelve cases (48%) reported intake of analgesics on < or = 8 days/month, five cases (20%) on 9-15 days/month and eight cases (32%) >15 days/month. One case (4%) reported on recurrent analgesic abuse. Outpatient (tapering analgesics gradually)--One case (5%) reported intake of analgesics on < or = 8 days/month and twenty-one cases (95%) reported daily intake. Fifteen cases (68%) reported continuous analgesic abuse. Comparison between migraine and tension-type headache suggested that patients with migraine showed a tendency towards a better prognosis than patients with tension-type headache. CONCLUSIONS: These results demonstrate the efficacy of withdrawal treatment in difficult cases suffering from analgesics abuse headache. If patients cannot be safely or adequately treated as outpatients, inpatient treatment may be needed. PMID- 16248394 TI - [Therapy of dystonia in Japan]. AB - A questionnaire about the treatment of dystonia was sent out to 585 councilors of Societas Neurologica Japonica. One hundred and sixty-eight replies (28.7%) were collected, although some of them were excluded from the analysis because of inappropriateness. 1) The number of patients previously experienced was < 10; 37 respondents (22.7%), 10-50; 83 (50.9%), 50-100; 26 (16.0%), and > 100; 17 (10.4%). 2) Oral medication was most often the first line treatment in either of generalized dystonia, blapharospasm, cervical dystonia, and writer's cramp. Botulinum toxin injection was the first or the second line treatment in 147 (87.5%) and 116 (69.0%) respondents for blepharospasm and cervical dystonia, respectively. In these two conditions, the more experienced doctors tended to prefer botulinum toxin injection to the other treatments as the first choice (Cochran-Armitage analysis; p = 0.003 for blepharospasm and p = 0.002 for cervical dystonia). 3) Among the oral drugs, anticholinergics, especially trihexyphenidyl, were the most frequent choice in generalized dystonia, cervical dystonia, and writer's cramp. For blepharospasm, clonazepam was most favored. Sedatives, especially diazepam, were also often the drug of choice in either of these disorders. The favored drugs were not related to the respondent's experience. 4) The success rate of treatment, designated as the percentage of patients who improved through any treatment so much that the respondent was satisfied with it, was the highest in blepharospasm (65.4 +/- 24.1; mean +/- SD), followed by cervical dystonia (41.2 +/- 23.4), writer's cramp (32.9 +/- 22.5), and generalized dystonia (20.4 +/- 19.8). Only in cervical dystonia, the rate was significantly higher in more experienced respondents (regression analysis; p = 0.008). In blepharospasm (p < 0.001) and cervical dystonia (p = 0.002), regression analysis indicated that the success rate was higher in the group who preferred botulinum toxin injection to oral medication as the first line treatment. These results indicate that in Japan the treatment of choice for dystonia does not always follow the therapeutic guidelines for dystonia proposed in some foreign countries. Adopting more evidence-based rationale of treatment is encouraged, because the recent progress about the treatment of dystonia, e.g. botulinum toxin injection or the stereotaxic surgery, is reshaping dystonia from a devastating to a treatable disorder. PMID- 16248395 TI - [Effect of intermittent positive pressure ventilation on life-span and causes of death in Duchenne muscular dystrophy]. AB - We have been introducing the intermittent positive pressure ventilation (IPPV) therapy in Duchenne muscular dystrophy (DMD) since 1990. Then, the changes of life-span and causes of mortality by IPPV were investigated in this study. The subjects were 157 patients of long-term followed up DMD in our hospital. The Kaplan-Meier's survival curve of 73 IPPV treated patients showed a median survival time of 31.0 years of age, whereas that of 20.4 years in 84 patients not treated by IPPV. The actual mean age of death in 29 IPPV treated patients was 25.68 +/- 5.18 years-old (M +/- SD), which was significantly higher than that in 74 patients with not-IPPV treated of 19.76 +/- 3.47. The IPPV treatment changed the major causes of death; 59.5% to 3.5% for respiratory failure, 12.2% to 37.9% for cardiac failure, and 0% to 10.3% for repiratory trouble. There were certain number of patients who died of repiratory infection, respiratory tract disorder, digestive organ disorders and sudden death. Thus, IPPV therapy is significantly effective for prolongation of life-span in DMD, and it seems to be necessary to establish the treatment strategy for cardiac failure and other potentially fatal complications. PMID- 16248396 TI - [Two cases of top of the basilar syndrome with onset seizure]. AB - We reported 2 patients with top of the basilar syndrome manifested by onset seizure. Patient 1 was a 76-year-old man. When he was sleeping, suddenly he gave a loud cry and went into convulsions. So he was brought to our hospital by ambulance. On admission, he had unconsciousness and left hemiplegia. Patient 2 was a 70-year-old man. When he was bathing, he lost his consciousness and brought to our hospital by ambulance. On admission, he had convulsion and tetraplegia. Both patients had convulsions and palsy when they had brain infarctions. Brain diffusion weighted MR image showed high intensity area in bilateral internal thalamus, brainstem and cerebellum and no high intensity area in cerebral cortex. We know well that Todd's palsy is palsy after convulsion, but vertebro-basilar occlusion also shows convulsion and palsy. Therefore attention should be paid in the case of the patients who had convulsion and palsy. PMID- 16248397 TI - [A case of acute cervical epidural hematoma mimicking medial medullary infarction complicated by arterial dissection]. AB - A 67-year woman with hypertension rapidly noted weakness of the right upper and lower extremities with posterior cervical pain. At admission to our hospital, she showed right hemiparesis without facial palsy, and bilateral pathological reflex. Chest X-ray demonstrated enlargement of the mediastinal shadow. We suspected that she developed aortic arch dissection extending into the vertebral arteries and subsequent medial medullary infarction. However, enhanced thoracic CT, brain MRI DWI and MRA examinations were negative. Her symptoms rapidly recovered three hours after the onset. A cervical MRI study revealed cervical epidural hematoma locating between the C3 and C6, which suppressed right side of the spinal cord. In conclusion, cervical epidural hematoma as well as medial medullary infarction complicated by the vertebral arterial dissection should be urgently explored when a patient had a sudden onset of cervical pain and hemiparesis without facial palsy. PMID- 16248398 TI - [A case of non-herpetic acute encephalitis with autoantibodies for ionotropic glutamate receptor delta2 and epsilon2]. AB - We report a 45-year-old woman admitted to our hospital due to fever, consciousness disturbance, and severe seizures. Based on her signs and symptoms and clinical course, a diagnosis of non-herpetic acute encephalitis was made. She received antibiotic drugs, acyclovir, gamma-globulin, and steroid pulse therapy (methylprednisolone 1 g/day, 3 days). The patient's signs, symptoms and severe seizure showed marked improvement, but she still showed monthly seizure attacks and both anterograde and retrograde amnesia Viral infection and autoimmune response after viral infection may have been involved in non-herpetic acute encephalitis in our patient. Recently, autoantibodies to GluRepsilon2 and VGKC were reported in cases of non-herpetic acute limbic encephalitis (NHALE). In our patient, we detected IgM type autoantibody to GluRdelta2 and epsilon2 in both the CSF and serum, and these antibodies normalized in the CSF with the clinical course. Autoantibodies to GluRdelta2 and epsilon2 may be involved in the clinical symptoms and pathogenesis of non herpetic acute limbic encephalitis. This is the first report of MRI-positive non-herpetic acute encephalitis with autoantibodies to GluRdelta2 and epsilon2. PMID- 16248399 TI - [Progressive multifocal leukoencephalopathy with idiopathic CD4 positive T lymphocytepenia mimicking a low grade glioma on proton MR spectroscopy. A case report]. AB - A 61-year-old man with no history of HIV infection developed a subacutely progressive dementia and left hemiparesis. Brain MRI showed a high intensity lesion in the right frontal lobe on T2 weighted image. There was no contrast enhancement after gadolinium-DTPA administration. 1H MRS revealed a marked decrease in the n-acetyl aspartate/creatine ratios and an increase in the choline/creatine ratio. A lactate peak also was present. A low-grade glioma was suspected and he was admitted to our hospital. On examination, there was a mild dementia and left hemiparesis. A peripheral blood count revealed lymphocytopenia (426/mm3) with a CD4/CD8 ratio of 0.28. No evidence of HIV infection, malignancies or collagen disease was found. A brain biopsy revealed no tumor cells but instead demyelinated brain tissue with large nucleated cells. JC virus antigen was detected in the cells of the demyelinated lesions. A diagnosis of PML associated with idiopathic CD4 positive lymphocytopenia was made. There are only a few reports concerning 1H-MRS findings in patients with PML and the present case illustrates the difficulty of making a differential diagnosis between PML and glioma. PMID- 16248400 TI - [A patient with coccidioidal meningoencephalitis]. AB - A 37-year-old man presented with coccidioidal meningoencephalitis (CM) 1 month after a preceding case of pneumonia. Initially, he could not be definitely diagnosed with CM because of nonspecific features of the clinical, laboratory, and radiological findings. However, we began to suspect CM because the patient had lived in endemic area of coccidioidomycosis, and our subsequent analysis provided evidence of complement-fixing antibodies for Coccidioides immitis in serum and CSF, leading us to a final diagnosis. The CM was intractable, despite intensive administration of fluconazole and amphotericin B. Although the patient's CM gradually and mildly improved, he also suffered from bacterial meningoencephalitis and left putaminal hemorrhage with intraventricular hematoma, which caused persistent right hemiparesis and dementia. The incidence of coccidioidomycosis in Japan is rapidly increasing. The initial clinical manifestation of coccidioidomycosis is usually pneumonia, which in most cases heals spontaneously. Coccidioidomycosis rarely presents with meningoencephalitis, which is thought to be fatal. Immediate and adequate initiation of anti-fugal treatment is necessary to obtain a better prognosis for CM. Careful history taking after a foreign trip is helpful when there is a suspicion of coccidioidomycosis. PMID- 16248401 TI - [A case of adult onset subacute sclerosing panencephalitis; sequential clinico radiological findings over two years]. AB - We present a very rare case of adult onset subacute sclerosing panencephalitis (SSPE), and explain the characteristic sequential clinico-radiological findings. The patient, a twenty three-year-old man, had noticed unsteadiness in walking about two months previously. Although inosine pranobex and intrathecal interferon were administered, symptoms worsened insidiously and he became bedridden after four months. The levels of serum and CSF anti-measles IgG antibodies have not changed. Initially, supratentorial cortical atrophy was observed, especially of the left temporal lobe, but there were no other MRI signal alterations at the time. After three months, the supratentorial cortices produced low-signal intensities in T1-weighted images and in T2-weighted MRI, the cortical margin was very unclear and white matter signal intensities had become higher. Furthermore, cortices became thinner and ventricular size increased, especially for the lateral and third ventricles. SPECT examinations showed a marked reduction in cerebral blood flow and the perfusion deficits observed seemed to be closely correlated with the abnormal MRI signal patterns. Pathological examinations of biopsy samples revealed infiltration of inflammatory cells around the small vessels. As for immunohistochemical findings, CD68 positive cells were frequently observed, and this result implied the activation of microglia. Further studies are necessary to elucidate the pathogenic mechanisms of SSPE. PMID- 16248402 TI - [A case of spontaneous intracranial hypotension with multiple CSF leaks in the thoracic region]. AB - A case of spontaneous intracranial hypotension (SIH) with multiple cerebrospinal fluid (CSF) leaks is reported. A 54-year-old man experienced a severe generalized headache associated with nausea and decreased hearing. The headache appeared when he was sitting or standing but was completely relieved by lying down. Cranial MRI with gadolinium infusion showed diffuse pachymeningeal enhancement. Spinal MRI demonstrated small amount of epidural fluid collection in the upper thoracic region. Radionuclide cisternography demonstrated CSF leaks at the Th2 and Th7 levels on the left side and at the Th3 through Th5 levels on the right side. Since bed-rest and intravenous transfusion for 10 days showed no beneficial effects, epidural blood patch (EBP) at the Th6/7 interspace was performed. After receiving EBP four times repeatedly, his symptoms entirely disappeared. SIH patients with multiple CSF leaks are rare in the literature. Although no consensus exists on the treatment strategy for such cases, our case suggests that at least 2 EBPs should be performed for each leak site before considering surgical treatment. PMID- 16248403 TI - [Progress of research on dementia with Lewy bodies]. PMID- 16248404 TI - [Establishment of brain bank for aging research]. AB - We have established a brain bank for the prevention and treatment of aging related movement and cognitive disturbances, as a joint project between a rural care hospital and a research institute. The resources of the bank are to be used for collaborative studies approved by the bank's committee. The collaborative investigators should also be qualified by the institute to conduct the research jointly. The collaborative studies require authorization by the institutional review board (IRB) of the institute, the hospital and each facility involved in collaborative studies. The bank continues to have the responsibility for the resources, after the transfer of the resources to the facilities of collaborative investigators, pursuant to Article 18 of the Cadaver Autopsy and Preservation Act Thus, the status of resource utilization and outcomes from their use in studies will be monitored periodically (every 6 months). We shared the philosophy with the brain banks in the United States that the resources of the bank, donated on the basis of a charitable spirit, belong to the public domain and are regarded as public resources to be used to contribute to promoting public welfare. PMID- 16248405 TI - [Chronic heart failure and sleep breathing disorders]. PMID- 16248406 TI - [Future aspect of geriatric medicine]. PMID- 16248407 TI - [Currently increasing cardiogenic cerebral embolism, especially in its prevention and treatment]. PMID- 16248409 TI - [Treatment of diabetes mellitus in the elderly]. PMID- 16248410 TI - [Vascular aging and oxidative stress]. PMID- 16248408 TI - [Metabolic syndrome and the management of traditional risk factors for ischemic heart disease in elderly people]. PMID- 16248411 TI - [Pathophysiological significances of citrullinated proteins in geriatric diseases]. PMID- 16248412 TI - [Respiratory disorders in the aged: in the present and future]. PMID- 16248413 TI - [Trainability in elderly]. PMID- 16248414 TI - [Problems associated with advancing Parkinson's disease]. PMID- 16248415 TI - [Cerebrovascular disease in the elderly]. PMID- 16248416 TI - [The treatment of atrial fibrillation--recent topics on the upstream therapy of atrial fibrillation]. PMID- 16248417 TI - [Supporting the elderly with dimentia and their family in the community]. PMID- 16248418 TI - [National survey on the current status of programs to teach end-of-life care to undergraduates of medical and nursing schools in Japan]. AB - AIM: We conducted a national survey to examine the current status of programs to teach end-of-life care to undergraduates of Japanese medical and nursing schools in 2004. METHODS: Our survey focused on the following areas: (1) the present status of curricula to teach end-of-life care, (2) topics covered in the curricula, (3) details of the timing of existing end-of-life care teaching programs and of departments responsible for it, (4) teaching methods, (5) education assessment tool, (6) reading list of textbooks. RESULTS: 50.6% of the medical schools and 40.9% of the nursing schools participated. Most of the schools offered end-of-life care education programs, and 45% of the medical schools and 68.9% of the nursing schools offered education concerning end-of-life care for elderly. The most frequent teaching timing was the fourth year of medical school and the third year of nursing school. Broader end-of-life topics were covered in the curriculum in the nursing schools than the medical schools. One fourth of the nursing schools offered a separate course in end-of-life care. The mean number of teaching hours was 7.6 in the medical schools and 35.5 in the nursing schools. More nursing schools offered educational programs in which students can gain experience with end-of-life care than medical schools. A few institutions used practiced examinations to evaluate students' learning. A reading list of end-of-life care textbooks was provided in 10% of the medical schools and 35.6% of the nursing schools. CONCLUSION: Our survey suggested that systematizing end-of-life care education and improvement in text content are on the way. PMID- 16248419 TI - [Prognosis of patients with myasthenia gravis]. AB - AIM: To study the correlation between therapy and outcome of patients with late adult onset and ocular type myasthenia gravis (MG). METHODS: The 58 MG patients admitted to our hospitals from 1991 to 2003 were classified into juvenile (younger than 20), early adult onset (20-64) and late adult onset (65 or older) groups, or also ocular and generalized types. We evaluated their therapeutic outcomes. RESULTS: The late adult onset group consisted of 11 patients, classified as 4 ocular and 7 generalized types. There were more women than men in both the late and early adult onset groups. Anti-acetylcholine receptor (anti AchR) antibodies were positive in all patients. Four patients had thymus lesions. Ten patients received treatment as follows: steroid therapy in 6, thymectomy in 4 and only choline-esterase (ChE) inhibitor therapy in 4 patients. However 4 patients worsened after treatment because of inadequate steroid therapy, 2 of whom experienced myasthenic crisis. On the other hand 21 patients were ocular type, of whom anti-AchR antibodies were positive in 12. They received treatments of the 21, steroid therapy was given to 12, thymectomy was performed in 4 and only ChE inhibitor therapy was given to 8 patients. One ocular type patient progressed to the generalized type. Six of 16 improved patients showed distinct recovery. CONCLUSION: In the late adult onset group careful assessment concerning timings of administration and reduction of steroid are required to avoid side effects and complications. One ocular type patient progressed to the generalized type and most patients had a good outcome, which might be related to steroid therapy. PMID- 16248420 TI - [Analysis of factors for progression of diabetic renal failure in elderly people]. AB - AIM: To investigate the factors in the progression of renal failure in type 2 diabetes in elderly people. MATERIALS AND METHODS: The subjects were 2034 patients hospitalized for type 2 diabetes therapy at Osaka City University Hospital from 1993 to 2000. Among them, 362 patients had diabetic non-dialyzed renal failure. Moreover, 93 patients reached the endpoint of doubling primary serum creatinine values. We analyzed the factors of progression for diabetic renal failure, and divided the cohorts into two groups. One group consisted of patients 65 years old or more (higher age group, HP), and the other of patients under 65 years old (lower age group, LP). We used the Kaplan-Meier methods and Cox proportional hazard models for the analysis of the cohorts with the endpoint of doubling the serum creatinine values. RESULTS: There were no significant differences between HP and LP by the Kaplan-Meier methods for progression of renal failure. According to Cox monofactorial-analysis, hypoalbuminemia was determined to be a significant factor in the progression of renal failure in both HP and LP. In the Cox multifactorial-analysis, hypoalbuminemia was a independent significant factor in HP, but not in LP. CONCLUSION: Hypoalbuminemia is considered to be a significant factor in the progression of renal failure in type 2 diabetes regardless of age, and may be a more significant factor in elderly people. PMID- 16248422 TI - [Better control of blood sugar with treatment using half-solid nutrients: a case report]. AB - We report on a 60-year old man who became bedridden after a cerebral infarction and was put on a regimen of percutaneous endoscopic gastrostomy (PEG) feedings. He developed such problems as intractable and prolonged diarrhea and leaking of nutrients from his gastric fistula, and also required frequent aspiration of his sputum. We then began treatment using half-solid nutrients. As a result, the diarrhea and leakage disappeared and the number of aspirations performed decreased. This patient has diabetes mellitus, with a high blood sugar level after feeding. After using half-solid nutrients, the peak became lower and appeared later, which is the same pattern seen with immuno-reactive insulin (IRI). This suggests that half-solid nutrients are useful in terms of blood sugar control. PMID- 16248421 TI - ["Can high fluid intake prevent cerebral and myocardial infarction?" Systematic review]. AB - OBJECTIVES: We performed a systematic review about whether high fluid intake can prevent cerebral and myocardial infarction. MATERIALS AND METHODS: Previously published papers were searched for in PubMed using the combined terms of dehydration, hydration, water intake, fluid intake, cerebral infarction, cerebrovascular disease, apoplexy, myocardial infarction, angina pectoris, ischemic heart disease, blood viscosity and hemorheology. RESULTS: Of 611 papers searched, twenty-two were selected. There was one prospective randomized study, four prospective non-randomized studies, eight epidemiologic (cohort or case control) studies and nine retrospective descriptive studies, presenting the following points. Dehydration, which increases blood viscosity, is one of the causes of cerebral or myocardial infarction. Important factors other than dehydration can cause an increase in viscosity. Drinking water during the night can protect an increase in blood viscosity but there has been no evidence that drinking excessive amount of water prevents cerebral infarction. There was one report that the risk of myocardial infarction was lower in people drinking more than 5 glasses of water than those drinking less than 2. CONCLUSION: Since cerebral and myocardial infarction are primarily caused by atherosclerosis and atheroma plaque, it is essential to adjust life style for prophylaxis. There has been no direct evidence that decrease in viscosity due to high fluid intake can prevent cerebral infarction. Further studies regarding the relationship between fluid intake and ischemic diseases, and the appropriate fluid intake for the elderly to improve their QoL are needed. PMID- 16248423 TI - [A case of small cell lung cancer with intramedullary spinal cord metastasis]. AB - An 83-year-old man was admitted with paraplegia and loss of all sensation below the level of umbilicus, with bowel and bladder dysfunction. Stage IV small cell lung cancer had been diagnosed two years ago and had received several courses of chemotherapies. A magnetic resonance imaging revealed an enhanced mass in the intramedullary spinal cord at the level of Th10-L1. Metastatic spinal tumor was diagnosed by clinical and radiological examinations. This is a rare case of small cell lung cancer with intramedullary spinal cord metastasis which caused various neurological symptoms. PMID- 16248424 TI - The transfection of embryonic stem cells with Tet-on system and its responsiveness to doxycycline. AB - We transiently transfected pTet-on and pTRE2hyg-luciferase into the mouse embryonic stem cells (ESCs) using lipofectamine, and analyzed its inductive effect by adding serial concentrations of doxycycline (DOX). The results showed that in the transfected group, the luciferase activity of the cells was gradually increased along with the increasing concentration of DOX. While in the non transfected group, the luciferase activity was not detectable even with DOX treatment. This indicated that the ESCs transfected with Tet-on system could response to DOX very well, and the regulation of target gene expression is dose dependent. PMID- 16248425 TI - "Beijing Region" (3pter-D3S3397) of the human genome: complete sequence and analysis. AB - The goal of the Human Genome Project (HGP) is to determine a complete and high quality sequence of the human genome. China, as one of the six member states, takes a region between 3pter and D3S3397 of the human chromosome 3 as its share of this historic project, referred as "Beijing Region". The complete sequence of this region comprises of 17.4 megabasepairs (Mb) with an average GC content of 42% and an average recombination rate of 2.14 cM/Mb. Within Beijing Region, 122 known and 20 novel genes are identified, as well as 42607 single nucleotide polymorphisms (SNPs). Comprehensive analyses also reveal: (i) gene density and GC content of Beijing Region are in agreement with human cytogenetic maps, i.e. G minus bands are GC-rich and of a high gene density, whereas G-plus bands are GC poor and of a relatively low gene density; (ii) the average recombination rate within Beijing Region is relatively high compared with other regions of chromosome 3, with the highest recombination rate of 6.06 cM/Mb in the subtelomeric area; (iii) it is most likely that a large gene, associated with the mammary gland, may reside in the 1.1 Mb gene-poor area near the telomere; (iv) many disease-related genes are genetically mapped to Beijing Region, including those associated with cancers and metabolic syndromes. All make Beijing Region an important target for in-depth molecular investigations with a purpose of medical applications. PMID- 16248426 TI - Construction of a novel kind of expression plasmid by homologous recombination in Saccharomyces cerevisiae. AB - Based on a previously used plasmid pHC11, a new plasmid pHC11R was constructed. Cutting plasmid pHC11R with proper restriction enzymes, the resulting larger DNA fragment pHC11R' was co-transformed with a PCR amplified expression cassette of human IFNalpha2b into yeast. By means of the homologous sequences at both ends of two DNA fragments, a novel expression plasmid pHC11R-IFNalpha2b was formed via homologous recombination in the yeast. Compared with pHC11-IFNalpha2b, the expression plasmid pHC11R-IFNalpha2b was smaller in size and in absence of antibiotic resistant gene. The stability and copy number of pHC11R-IFNalpha2b were greatly increased and the expression level of heterologous protein was improved. As the derivatives of pHC11R, a series of recombination expression vectors pHRs containing different combination of expression elements were developed. This led to a rapid and powerful method for cloning and expressing of different genes in yeast. PMID- 16248427 TI - Functional genomics of maize submergence tolerance and cloning of the related gene Sicyp51. AB - In this study, SSH (Suppression Subtractive Hybridization) and cDNA microarray were used to identify genes associated with waterlogging response of maize roots. Mo17 and Hz32 are two maize inbred lines with differential tolerance to hypoxia. Seedlings of the inbred lines with two leaves were submerged in hypoxia buffer. SSH libraries were constructed with cDNA samples from roots. Both forward and reverse subtractions were performed for each inbred line, and 105 positive clones induced by hypoxia were selected by differential screening. The treated and control message RNA were hybridized with the cDNA microarray of Mo17, sequentially, 57 of 3-fold differentially expressed clones were obtained. A total of 162 positive clones were all sequenced. Bioinformatics analysis showed these positive clones represent 85 TUGs, including genes involved in several biochemistry pathways, such as glycolysis, protection, signal transduction, cell construction and energy metabolism and 41 EST with unknown function. Comparison between Mo17 and Hz32 indicates that genes related to hypoxia tolerance have different expression patterns in submerged roots. Several positive clones' expression patterns were revealed by Northern or RT-PCR, and a new gene (Sicyp51), which may contribute to hypoxia tolerance, was identified. PMID- 16248428 TI - Gene cloning and expression of cadherin in midgut of Helicoverpa armigera and its Cry1A binding region. AB - Cadherins belong to one of the families of animal glycoproteins responsible for calcium-dependent cell-cell adhesion. Recent literatures showed that the cadherin like in midgut of several insects served as the receptor of Bt toxin Cry1A and the variation of cadherin-like is related to insect's resistance to Cry1A. The full-length cDNA encoding cadherin-like of Helicoverpa armigera is cloned by degenerate PCR and RACE techniques and the gene was designated as BtR-harm, which is 5581 bp in full-length, encoding 1730 amino acid residues (BtR-harm was deposited in GenBank and the accession number is AF519180). Its predicted molecular weight and isoelectric point were 195.39 kDa and 4.23, respectively. The inferred amino acid sequence includes a signal sequence, 11 cadherin repeats, a membrane-proximal region, a transmembrane region and a cytoplasmic region. Sequence analysis indicated that the deduced protein sequence was most similar to the cadherin-like from Heliothis virescens with 84.2% identity and highly similar to three other lepidopteran cadherin from Bombyx mori, Manduca sexta and Pectinophora gossypiella, with the sequence identities of 60.3.6%, 57.5% and 51.0%, respectively. The cDNA encoding cadherin gene was expressed successfully in E. coli and the recombinant proteins can bind with Cry1Ac. Truncation analysis and binding experiment of BtR-harm revealed that the Cry1A binding region was a contiguous 244-amino acid sequence, which located between amino acid 1217 and 1461. Semi-quantitative RT-PCR analysis showed that BtR-harm was highly expressed in midgut of H. armigera, very low expressed in foregut and hindgut and was not expressed in other tissues. After H. armigera producing resistance to Cry1Ac, the expression quantity of BtR-harm significantly decreased in midgut of H. armigera. It is the first confirmation that BtR-harm can function as receptor of Cry1Ac in H. armigera and the binding region was located on a contiguous 244 amino acid sequence, suggesting that the decrease of expression quantity of BtR-harm is one of the main reasons for H. armigera resistance to Cry1Ac. PMID- 16248429 TI - cDNA cloning, functional expression and cellular localization of rat liver mitochondrial electron-transfer flavoprotein-ubiquinone oxidoreductase protein. AB - A membrane-bound protein was purified from rat liver mitochondria. After being digested with V8 protease, two peptides containing identical 14 amino acid residue sequences were obtained. Using the 14 amino acid peptide derived DNA sequence as gene specific primer, the cDNA of correspondent gene 5'-terminal and 3'-terminal were obtained by RACE technique. The full-length cDNA that encoded a protein of 616 amino acids was thus cloned, which included the above mentioned peptide sequence. The full length cDNA was highly homologous to that of human ETF QO, indicating that it may be the cDNA of rat ETF-QO. ETF-QO is an iron sulfur protein located in mitochondria inner membrane containing two kinds of redox center: FAD and [4Fe-4S] center. After comparing the sequence from the cDNA of the 616 amino acids protein with that of the mature protein of rat liver mitochondria, it was found that the N terminal 32 amino acid residues did not exist in the mature protein, indicating that the cDNA was that of ETF-QOp. When the cDNA was expressed in Saccharomyces cerevisiae with inducible vectors, the protein product was enriched in mitochondrial fraction and exhibited electron transfer activity (NBT reductase activity) of ETF-QO. Results demonstrated that the 32 amino acid peptide was a mitochondrial targeting peptide, and both FAD and iron-sulfur cluster were inserted properly into the expressed ETF-QO. ETF-QO had a high level expression in rat heart, liver and kidney. The fusion protein of GFP ETF-QO co-localized with mitochondria in COS-7 cells. PMID- 16248430 TI - Cloning and expression of swine myostatin gene and its application in animal immunization trial. AB - We have amplified swine myostatin (MSTN) gene by reverse transcription polymerase chain reaction (RT-PCR) and cloned it into pGEM-T Easy vector. The cloned swine MSTN gene consists of 1128 nucleotides, which has been submitted to GenBank (acquired registered code--AY448008). The cloned swine MSTN gene was successfully expressed in E. coli without the first 25 amino acids. Crude extraction of the expressed recombinant MSTN protein was used to immunize mice to investigate the effects on their bodyweights. We show here that the body weights of the immunized mice were higher than that of the controls, even though the difference was not significant. Surprisingly, the progenies of the immunized mice also were heavier than the controls. Especially at day 3, the average body weight of the immunized mice was 10.5% higher than that of the controls, which is significant (p < 0.05). PMID- 16248431 TI - Evolutionary trace analysis of eukaryotic DNA topoisomerase I superfamily: identification of novel antitumor drug binding site. AB - The studies of novel inhibitors of DNA topoisomerase I (Topo I) have already become very promising in cancer chemotherapy. Identifying the new drug-binding residues is playing an important role in the design and optimization of Topo I inhibitors. The designed compounds may have novel scaffolds, thus will be helpful to overcome the toxicities of current camptothecin (CPT) drugs and may provide a solution to cross resistance with these drugs. Multiple sequence alignments were performed on eukaryotic DNA topoisomerase I superfamily and thus the evolutionary tree was constructed. The Evolutionary Trace method was applied to identify functionally important residues of human Topo I. It has been demonstrated that class-specific hydrophobic residues Ala351, Met428, Pro431 are located around the 7,9-position of CPT, indicating suitable substitution of hydrophobic group on CPT will increase antitumor activity. The conservative residue Lys436 in the superfamily is of particular interest and new CPT derivatives designed based on this residue may greatly increase water solubility of such drugs. It has also been demonstrated that the residues Asn352 and Arg364 were conservative in the superfamily, whose mutation will render CPT resistance. As our molecular docking studies demonstrated they did not make any direct interaction with CPT, they are important drug-binding site residues for future design of novel non-camptothecin lead compounds. This work provided a strong basis for the design and synthesis of novel highly potent CPT derivatives and virtual screening for novel lead compounds. PMID- 16248432 TI - Epstein-Barr virus-encoded latent membrane protein 1 modulates cyclin D1 by c Jun/Jun B heterodimers. AB - In our recent studies, we found that LMP1 encoded by Epstein-Barr virus could accelerate the formation of active c-Jun/Jun B heterodimer. We studied the regulation of cyclinD1 by c-Jun/Jun B heterodimers by laser scanning confocal influorescence microscopy, Western blot, luciferase activity assay, super-EMSA and flow cytometry in the Tet-on-LMP1 HNE2 cell line, in which LMP1 expression was regulated by Tet-on system. c-Jun/Jun B heterodimers induced by LMP1 could up regulate cyclin D1 promoter activity and expression. Overexpression of cyclin D1 accelerated the progression of cell cycle. PMID- 16248433 TI - A seqlet-based maximum entropy Markov approach for protein secondary structure prediction. AB - A novel method for predicting the secondary structures of proteins from amino acid sequence has been presented. The protein secondary structure seqlets that are analogous to the words in natural language have been extracted. These seqlets will capture the relationship between amino acid sequence and the secondary structures of proteins and further form the protein secondary structure dictionary. To be elaborate, the dictionary is organism-specific. Protein secondary structure prediction is formulated as an integrated word segmentation and part of speech tagging problem. The word-lattice is used to represent the results of the word segmentation and the maximum entropy model is used to calculate the probability of a seqlet tagged as a certain secondary structure type. The method is markovian in the seqlets, permitting efficient exact calculation of the posterior probability distribution over all possible word segmentations and their tags by viterbi algorithm. The optimal segmentations and their tags are computed as the results of protein secondary structure prediction. The method is applied to predict the secondary structures of proteins of four organisms respectively and compared with the PHD method. The results show that the performance of this method is higher than that of PHD by about 3.9% Q3 accuracy and 4.6% SOV accuracy. Combining with the local similarity protein sequences that are obtained by BLAST can give better prediction. The method is also tested on the 50 CASP5 target proteins with Q3 accuracy 78.9% and SOV accuracy 77.1%. A web server for protein secondary structure prediction has been constructed which is available at http://www.insun.hit.edu.cn:81/demos/biology/index.html. PMID- 16248434 TI - Comparative genomics and phylogenetic analysis of S. dysenteriae subgroup. AB - Genomic compositions of representatives of thirteen S. dysenteriae serotypes were investigated by performing comparative genomic hybridization (CGH) with microarray containing the whole genomic ORFs (open reading frames, ORFs) of E. coli K12 strain MG1655 and specific ORFs of S. dysenteriae A1 strain Sd51197. The CGH results indicated the genomes of the serotypes contain 2654 conserved ORFs originating from E. coli. However, 219 intrinsic genes of E. coli including those prophage genes, molecular chaperones, synthesis of specific O antigen and so on were absent. Moreover, some specific genes such as type II secretion system associated components, iron transport related genes and some others as well were acquired through horizontal transfer. According to phylogenic trees based on genetic composition, it was demonstrated that A1, A2, A8, A10 were distinct from the other S. dysenteriae serotypes. Our results in this report may provide new insights into the physiological process, pathogenicity and evolution of S. dysenteriae. PMID- 16248436 TI - Let's pay more attention to human factors. PMID- 16248435 TI - Analysis of cytochrome P450 genes in silkworm genome (Bombyx mori). AB - We have searched the Bombyx mori genome for members of the major enzyme family, the Cytochrome P450s, which carry out multiple reactions to enable organisms to rid themselves of foreign compounds. As a result, 86 putative P450s were discovered in silkworm genome, which are thought to belong to 32 subfamilies. A comparative genomic analysis with Drosophila melanogaster reveals that the two insects have some similar P450 distribution patterns but still have some obvious differences. Especially, the diverse distribution exists in 7 p450 subfamilies, which are CYP4A, CYP4C, CYP4D, CYP6A, CYP6AE, CYP6B and CYP9A. Furthermore, we collected expression sequence tag (EST) evidence for 49 putative P450s genes, which are expressed at the transcriptional level and more likely to be true P450s. PMID- 16248437 TI - Keep IT coming. PMID- 16248438 TI - Advisory warns of EMI to patient monitoring devices. PMID- 16248439 TI - How to build your role in healthcare construction projects. PMID- 16248440 TI - What you should know about software engineering. PMID- 16248441 TI - Is the warning effective? Clinical alarms remain an area for patient safety improvement. PMID- 16248442 TI - Illumination in the operating room. PMID- 16248443 TI - A view from the trenches...cellular phone use in hospitals. PMID- 16248444 TI - Plethysmography. PMID- 16248445 TI - Managing patient monitoring network traffic. PMID- 16248446 TI - In-house software package helps close the loop on safety notifications. PMID- 16248447 TI - Dialing for jobs: how to make the most of a phone interview. PMID- 16248448 TI - Safeguards are paramount. PMID- 16248449 TI - Uses and misuses of probability in medical device risk management. PMID- 16248450 TI - Industrial techniques help reduce hospital-acquired infection. PMID- 16248451 TI - A new single-use bone marrow biopsy needle. AB - A new needle for taking a bone marrow biopsy sample from the posterior iliac crest is described. The needle comprises a hollow tube having a front-end portion formed to a reduced diameter. The front end is tapered by means of circumferentially spaced facets, forming a cutting edge. A series of flutes/serrations are introduced at the tapering transitional portion, between the main wider portion of the hollow tube and its reduced-diameter front-end portion. This design is new and is not available in any other conventional biopsy needle. These serrations help in its penetration through the hard cortical bone. A spacer guide, which is placed between the plastic dome of the needle and the stilette cap to enhance the ergonomics, is also new. The inside of the spacer guide is funnel shaped, through which the stilette is passed, thereby ensuring an easy and accurate alignment of the distal ends of the hollow needle and the stilette. This technique facilitates removal of the biopsy sample rearwardly out of the hollow needle particularly when the mouth (distal-end opening) of the needle is covered by extraneous bony soft tissue. In this technique, the same stilette that occupies the needle during its entry into the marrow cavity is used in sample removal. Thus, the need for an additional pusher to remove the biopsy sample from within the lumen of the needle has been eliminated. PMID- 16248452 TI - Evaluation of body composition methods for accuracy. AB - The objective of this study was to evaluate the accuracy of a variety of portable methods and instruments used to estimate body composition or percentage body fat (%BF) in a systematic, comprehensive manner on a wide range of subjects. The %BF was estimated using four skinfold protocols, three girth measurement protocols, two bioelectric impedance analysis (BIA) instruments, and one near-infrared instrument on 121 subjects. The subjects ranged in age from 21 to 51 years; weight ranged from 105 to 226 pounds and %BF from 8.3% to 38.3%; and the group was 29% male. The %BF estimates were compared to the values obtained from the generally accepted reference standard, underwater weighing (UWW). The correlation coefficients (r) between the test methods and UWW ranged from 0.48 to 0.72. Regression analysis resulted in a range of slopes from 0.48 to 0.93, y-intercept range from 3.8 to 13.1, and standard error of the estimate range from 3.8 to 7.5. All of the methods tended to overestimate lower and underestimate higher %BF. With two exceptions, all methods appeared more accurate for males than females. In general, the near-infrared appeared least and BLA appeared most accurate. In conclusion, in our opinion, most of the methods were not sufficiently accurate to use on a wide range of individuals. Although they may be sufficiently accurate on the narrow sample of subjects on which they were developed, they may not be as accurate for the general population. PMID- 16248453 TI - Sometimes, you can catch lightning in a bottle. PMID- 16248454 TI - Preventive maintenance redefined. PMID- 16248455 TI - [The bacteria isolated from middle nasal meatus and ethmoid sinus of chronic naso sinusitis patients and their correlationship]. AB - OBJECTIVE: To explore the bacteria isolated from middle nasal meatus and ethmoid sinuses of chronic naso sinusitis patients and their correlationship. METHOD: Patients with chronic naso sinusitis were selected to performed the functional nose endoscopy in order to obtain the sample with disinfection cotton. All the samples were cultured aerobically and anaerobically, separately. RESULT: In 120 samples of chronic naso sinusitis patients, 2 section of 18 samples (15%) showed the same kinds of bacteria culture positive; and 2 section of 9 samples showed different kinds of bacteria, 24 samples showed culture positive only in the samples from middle nasal meatus; and 12 samples showed culture positive only in the samples from ethmoid sinuses. The positive rate of isolation was 42.5% in middle nasal meatus and 32.5% in ethmoid sinuses. The positive rate of aerobe was 39.2% in middle nasal meatus, and 22.5% in ethmoid sinuses. The positive rate of anaerobe was 12.5% in middle nasal meatus and in ethmoid sinuses. CONCLUSION: The positive rate of isolation was not high in the sample with disinfection cotton of chronic naso sinusitis patients. The correlationship of bacterium between the middle nasal meatus and ethmoid sinuses is little. PMID- 16248456 TI - [Expression of ECP during different phrase of transitionary process of mucosa in sinus cavity after endoscopic sinus surgery]. AB - OBJECTIVE: To study the secretion of ECP during different phrase of transitionary process of mucosa in sinus cavity after endoscopic sinus surgery. METHOD: Secretion of ECP was determined with enzyme-fluoroimmunoassay method during different phrase of transitionary process of mucosa in sinus cavity in 20 patients with type I and type II chronic sinusitis after endoscopic sinus surgery. RESULT: Comparing the patients in 1-2 week after surgery with the conditions before surgery,a significant decrease of secretion of ECP was found (P < 0.01). In 4-6 weeks after surgery, increasing amounts of secretion of ECP were found, nearly reaching the conditions before surgery again. In 10-12 weeks, the secretion of ECP tended to the level as 1-2 week after surgery. CONCLUSION: This study may indicate that the ECP involves in the transitionary process of mucosa in sinus cavity after endoscopic sinus surgery. PMID- 16248457 TI - [Serum levels and clinical values of soluble intercellular adhesion molecule-1 in patients with nasal polyps]. AB - OBJECTIVE: To assess the role of soluble intercellular adhesion molecule-1 (sICAM 1) in etiology of nasal polyps. METHOD: Twenty-nine patients with nasal polyps and 20 healthy control cases were chosen. Their serum sICAM-1 was determined by a sandwich enzyme-linked immunosorbent assay (ELISA). All patients were treated by glucocorticoid, serum sICAM-1 was determined post-treatment again. RESULT: (1) Serum levels of sICAM-1 in patients with nasal polyps were higher than that of control group: (2) Serum levels of sICAM-1 in patients were reduced after glucocorticoid treatment, the decrease of sICAM-1 was more apparent in evident effect group than in effect group; (3) There was a close correlation between the decrease of polyp-size scores and the decrease of serum sICAM-1. CONCLUSION: The elevated expressions of sICAM-1 in patients with nasal polyps indicate its important role in eosinophil recruitment, which was considered as a representative hallmark for nasal polyps. The sICAM-1 is down-regulated by systemic and topical glucocorticoid treatment and the decrease of sICAM-1 is correlated with clinical effect. To take these lines of evidence together, anti adhesion therapy may be effective for nasal polyps, serum sICAM-1 levels may be use to monitor the effect of steroid therapy in nasal polyps. PMID- 16248458 TI - [The protein expression difference of transforming growth factor beta1, matrix metalloproteinases 1,7,9 and tissue inhibitors of matrix metalloproteinases-1 between chronic rhinosinusitis, nasal polyps and normal mucosa tissues]. AB - OBJECTIVE: To compare the expression difference of transforming growth factor beta1 (TGF-beta1), matrix metalloproteinases1,7,9 (MMP-1,7,9 ) and tissue inhibitors of matrix metalloproteinases-1 (TIMP-1) protein among the chronic rhinosinusitis tissues, nasal polyps tissues and normal nasal mucosa tissues. METHOD: The protein expression of TGF-beta1, MMP-1, MMP-7, MMP-9 and TIMP-1 in chronic rhinosinusitis tissues from 22 patients, in nasal polyps tissues from 21 patients and in inferior turbinate tissues from 15 patients underwent nasal septum operation was detected by enzyme-linked immunosorbent assay (ELISA). RESULT: (1) In chronic rhinosinusitis and nasal polyps tissues, compared with controls, the expression of TGF-beta1, MMP-7, MMP-9, TIMP-1 protein was significant increased (P < 0.05); (2) The expression of TGF-beta1 and TIMP-1 in chronic rhinosinusitis tissues was significant increased versus that in nasal polyps tissues (P < 0.05); (3) Concentration of MMP-7 protein in nasal polyps tissues was found significant higher than that in chronic rhinosinusitis tissues (P < 0.01). CONCLUSION: Different TGF-beta1, MMP-7, TIMP-1 protein level in chronic rhinosinusitis and nasal polyps tissues might be the molecular basis of the different histopathological character of chronic rhinosinusitis and nasal polyp tissues. PMID- 16248459 TI - [Surgical management of nasal septum malignant tumor]. AB - OBJECTIVE: To summarize the surgical treatment strategy of nasal septum malignant tumor. METHOD: Between 1993 October and 2003 February, 16 patients with nasal septum malignant tumor were treated. In a retrospective review, the pathological character, the operation reports, and the follow up data were analyzed, particularly with respect to the surgical approaches. RESULT: Ten male, 37-79 years old, average 52.2 years old; 6 female, 18-71 years old, average 58 years old. The pathology of nasal septum malignant tumor in our department were: seven cases with squamous cell carcinoma in (5 cases were primary, 2 cases were secondary), 4 cases with adenoid cystic carcinoma (3 cases were primary, 1 case was secondary), 3 cases with malignant melanoma, 1 case with adenocarcinoma, 1 case with mucoepidermoid carcinoma. Duration range was from 1 month to 7 months. The strategy of operation was that 8 cases with lateral rhinotomy, 5 cases with endoscopy, 2 cases with midfacial degloving, 1 case via uvular pathway. All patients received postoperation radiation (30 Gy). After follow up for 1-10 years, 1 case was undetected, 2 cases were dead (1 case with malignant melanoma, 1 case with secondary squamous cell cancer), 3 cases survive with tumor (1 case with squamous cell cancer, 1 case with secondary adenoid cystic cancer, 1 case with malignant melanoma). Ten cases survive without tumor. The rate of death, survival with tumor and survival without tumor is 12.5%, 18.8%, 62.5%, respectively. CONCLUSION: The prognosis of nasal septum malignant tumor is related to operation, radiation and pathological character. The surgical approach to nasal septum malignant tumor is related to the size of the tumor and the age of patient. It classifies the following indications: (1) Endoscopic approach in cases with tumor localized in nasal septum; (2) Lateral rhinotomy in cases with extensive tumor infiltration of the nasal septum, nasal turbinates, or even sinus; (3) Midfacial degloving or via uvular pathway in cases who are young persons. PMID- 16248460 TI - [Nasal NK/T cell lymphoma: a report of 11 cases]. AB - OBJECTIVE: To investigate the clinical diagnosis consideration of nasal NK/T cell lymphoma. METHOD: Reviewing the clinical data of 11 patients with nasal NK/T cell lymphoma between 1992 to 2003. RESULT: Eight patients were unilateral, 3 bilateral. Six patients were misdiagnosed of other diseases. Three were diagnosed correctly through pathological biopsy once. Eight patients died in 3 to 20 months. CONCLUSION: Nasal NK/T cell lymphoma is easily misdiagnosed. The right diagnosis can be built with clinical, pathological and imaging examination. PMID- 16248461 TI - [Study on the expression of HSP70 and HSP90beta in nasopharyngeal carcinoma and the clinical significance]. AB - OBJECTIVE: To study the expression of HSP70 and HSP90beta and its clinical significance in human nasopharyngeal carcinoma. METHOD: Immunohistochemical method SP was used to detect the expression of HSP70 and HSP90beta in 50 cases and its clinical significance were studied. RESULT: The positive rate of HSP70 and HSP90beta were 72% and 56% respectively. Analysis of patients, survival demonstrated that the prognosis of NPC with HSP70 (-) and HSP90beta (-) expression were significantly better than the others. CONCLUSION: The overexpression of HSP70 and HSP90beta were probably concerned with the occurrence, development and prognosis of nasopharyngeal carcinoma. PMID- 16248462 TI - [Study on expression and relationship between proliferating cell nuclear antigen and p53 oncoprotein in nasopharyngeal carcinoma]. AB - OBJECTIVE: To observe PCNA p53 gene expression and their relationship with clinicopathologic factors in primary nasopharyngeal carcinoma(NPC). METHOD: PCNA, p53 proteins were assayed with immunohistochemical technique. RESULT: The expression of p53 protein was 9.5% in chronic inflammation of nasopharyngeal epithelium(CINE), which was significantly lower than it in NPCs with 76.3% (P < 0.01). In NPCs. p53 gene expression was significantly correlated with cervical lymph node metastasis of NPCs (P < 0.01), The expression of p53 in stage II, III, lV (93.3%, 80.0% and 100% respectively) were significantly higher than that in stage I (40.0%) (P < 0.01), the expression of PCNA and p53 protein was highly correlated (P < 0.01). CONCLUSION: The expression of p53 is the useful factor in diagnosis of NPC, the overexpression of p53 is related with the stage and cervical lymph node metastasis of NPC. And PCNA PI may represent the proliferative state of NPC, all of them may play important roles in the pathogenesis of nasopharyngeal carcinoma. PMID- 16248463 TI - [Effects of bFGF on the nasal mucosa after endoscopic sinus surgery]. AB - OBJECTIVE: To explore the effect of bFGF on nasal mucosa after ESS. METHOD: Ten cases of type I phase 3 chronic sinusitis were selected randomly. For each patient bFGF was intranasally used in one single nasal cavity a week after ESS. The other cavity, treated with nothing, was used as control. Scanning electron microscopy examinations of middle meatus mucosa were made 1,1. 5, 2 and 3 month after ESS. RESULT: In the first month after operation ciliated cells in the mucosa swelled and lined unregularly, the cilia exfoliated. From the sixth week to the 2nd month ciliated cells lined more regularly and the number of cells as well as the density of cilia increased obviously but the cilia aligned in disturbance. In the 3rd month after ESS, the swell of mucosa subside completely and cilia of columnar epithelial cells moved in one direction,which meant the cilia recover the function. CONCLUSION: bFGF can speed up the recovery of nasal mucosa both in number and in function after ESS. PMID- 16248464 TI - [Expression of six gene proteins in laryngeal carcinomas and their significance]. AB - OBJECTIVE: To investigate the expression of p15, p16, Rb, p27,p53 and PCNA gene proteins in different clinical stage, different position and different histological types of laryngeal carcinomas and their significance. METHOD: p15, p16, Rb, p27, p53 and PCNA gene proteins were examined with immunohistochemical SP method in the paraffin-embedded specimens of 65 cases with LSCC and 30 cases of normal or proliferative mucosas of larynx as a contrast. The result was analyzed by chi-square test. RESULT: The expression of the six gene proteins in LSCC and in laryngeal with normal or proliferative mucosas was different, the difference was significant (P < 0.01); the expression of p15 and p27 was significantly related with the position/clinical stage/histological types of LSCC (P < 0.01); the expression of p16 was significantly negative but the PCNA was positive correlated with th histological types of LSCC (P < 0.01). Two of them did not correlate with the position or clinical stage of LSCC (P > 0.05). There was trend of the decreasing Rb expression or increasing p53 expression with increasing clinical stages of LSCC, and the expression of p53 was also positive correlated with the histological types of LSCC. CONCLUSION: p15 and p27 play an important role in the development and progression of LSCC, and can be used as markers for assessment the treatment and prognosis of LSCC. The expression of p16 and PCNA can reflect the proliferation activity in the tumor cells, and is correlated with the development of tumor, but it does not correlate with the progression of tumor. The expression of Rb and p53 were more correlated with the progression of tumor. PMID- 16248465 TI - [Culture and identification of microvascular endotheliocytes from human nasal inferior turbinate tissue]. AB - OBJECTIVE: To study the method of culturing human nasal inferior turbinate endotheliocytes in vitro, and to provide the study base for nasal-related disease. METHOD: The endotheliocytes from human nasal inferior turbinate were gotten in nasal surgery and cultured in vitro. RESULT: We got human nasal inferior turbinate microvascular endotheliocytes in vitro, and confirmed them by immunity fluorescence. CONCLUSION: Human nasal inferior turbinate microvascular endotheliocytes can be cultured in vitro, and be used in the studies of nasal related disease. PMID- 16248466 TI - Differential expression of muscle damage in humans following acute fast and slow velocity eccentric exercise. AB - We sought to determine if the velocity of an acute bout of eccentric contractions influenced the duration and severity of several common indirect markers of muscle damage. Subjects performed 36 maximal fast (FST, n = 8: 3.14 rad x s(-1)) or slow (SLW, n = 7: 0.52 rad x s(-1)) velocity isokinetic eccentric contractions with the elbow flexors of the non-dominant arm. Muscle soreness, limb girth, plasma creatine kinase (CK) activity, isometric torque and concentric and eccentric torque at 0.52 and 3.14 rad x s(-1) were assessed prior to and for several days following the eccentric bout. Peak plasma CK activity was similar in SLW (4030 +/ 1029 U x 1(-1)) and FST (5864 +/- 2664 U x 1(-1)) groups, (p > 0.05). Both groups experienced similar decrement in all strength variables during the 48 hr following the eccentric bout. However, recovery occurred more rapidly in the FST group during eccentric (0.52 and 3.14 rad x s(-1)) and concentric (3.14 rad x s( 1)) post-testing. The severity of muscle soreness was similar in both groups. However, the FST group experienced peak muscle soreness 48 hr later than the SLW group (24 hr vs. 72 hr). The SLW group experienced a greater increase in upper arm girth than the FST group 20 min, 24 hr and 96 hr following the eccentric exercise bout. The contraction velocity of an acute bout of eccentric exercise differentially influences the magnitude and time course of several indirect markers of muscle damage. PMID- 16248467 TI - The magnitude of muscle damage induced by downhill backward walking. AB - While various models for exercise-induced muscle damage (EIMD) have been introduced, many of them use maximal voluntary contractions of the elbow flexors and knee extensors performed on isokinetic dynamometers. Few studies have used exercise protocols that attempt to replicate submaximal eccentric muscle actions that commonly occur during daily activities. Downhill backwards walking has been used previously as an EIMD model. However, the common markers of muscle damage have not been systematically examined for this model. The purpose of this study was to determine the magnitude of muscle damage induced by downhill backward walking with regard to changes in commonly-used indirect markers of EIMD. Twenty subjects aged between 19 y and 42 y completed a bout of 60 min of downhill (-15%) backward walking in which a single limb performed submaximal eccentric actions at a stepping rate of 30 - 35 strides per min. A repeated measures ANOVA revealed significant (p < 0.05) increases from baseline for soreness (24 hr- 96 hr), tenderness (24 hr - 96 hr), and plasma creatine kinase activity (0.5 hr - 96 hr), and significant decreases (p < 0.05) in maximal voluntary isometric (approximately 25%) and isokinetic (-15%) strength (0.5 hr - 96 hr) post-walk for the exercised limb. The time course of observed changes in these markers was similar to that reported for EIMD models of the elbow flexors and knee extensors. However, the magnitude of muscle damage appeared more consistent with that demonstrated following submaximal eccentric exercise. PMID- 16248468 TI - Resistance training effects on muscular strength of elderly are related to intensity and gender. AB - The purpose of this study was to determine whether a high intensity (HI) versus a moderate (MI) or low-intensity (LI) training program would be more effective in improving the isokinetic knee extension muscular performance in healthy inactive men and women. Sixty-four participants, men and women, were randomly assigned to one of four groups: control group (C), LI (50% of 1RM), the MI group (70% of 1RM) and the HI (90% of 1RM). Participants exercised on three resistance exercise machines: leg extension, leg curls and leg press. The isokinetic testing method (concentric mode) applied prior to and at the end of the training period (16 weeks, three 3 times per week) to assess the knee muscular performance. MANOVA repeated measures revealed that the HI group demonstrated the most strength gains following a speed specificity pattern (most considerable improvement occurred at or near slow speeds from 7.3% to 11.2% for male and from 2.3% to 15.2% for female). In addition, males demonstrated a greater improvement of knee extension power output than females. In conclusion, HI strength training is proposed for elderly men and women as the most effective protocol. Furthermore only at low velocity testing, women of the HI showed a greater change than men (p < 0.05). Regarding strength increase in relation to various testing velocities, a greater increase was found in HI at low velocities, with the other training groups exhibiting almost similar strength increase at all tested speeds. PMID- 16248469 TI - Methods of the NSW Schools Physical Activity and Nutrition Survey (SPANS). AB - The NSW Schools Physical Activity and Nutrition Survey (SPANS 2004) (N = 5407) was a representative population survey, conducted February-May 2004, among New South Wales (Australia) school students aged 4 y-16 y. Physical activity, fitness, fundamental movement skill proficiency and food habits are all related to overweight and obesity and were a focus of this study. The last population based survey in NSW was conducted in 1997 and use of the same measurement instruments (with the exception of food habits) allowed examination of secular trends. Overweight and obese children are at risk for a number of serious, long term health problems, and a sub-sample (n = 500) of urban dwelling Year 10 students (mean age 15.4 years) gave a fasting blood sample which was analysed for the presence of risk factors for the metabolic syndrome. This paper describes the methods of the NSW Schools Physical Activity and Nutrition Survey 2004 (SPANS 2004). PMID- 16248470 TI - Promoting respect for the rules and injury prevention in ice hockey: evaluation of the fair-play program. AB - OBJECTIVE: To reduce the number of transgressions to the rule, the occurrence of violent acts and to prevent injuries, Hockey Quebec adopted the Fair-Play Program (FPP). The objective of the present study was to evaluate the effectiveness of the FPP. METHODS: 52 Bantam (14-15 years) teams participated in this cohort study. In total, 49 games (13 with the FPP, 36 without FPP) were systematically assessed for transgressions to the rule. Body checking was allowed in all games. Transgressions to the rule data were obtained using a real time observation system in a natural setting, while injury data were collected through a self administered questionnaire. Data were analysed using generalised linear models with generalised estimating equations accounting for potential team effect. RESULTS: The number of penalties per game was significantly lower (p < 0.01) for games played with the FPP. Overall, no difference was noted in the number of transgressions observed during games played with or without the FPP. Players in leagues where the FPP was used held their opponents more frequently (p < 0.0001). On the other hand, players in leagues without the FPP shoved and hit more (p = 0.05). No difference was noted in the injury rate for games played with or without the FPP. CONCLUSIONS: This study showed that the FPP is one of the tools available to help those in the hockey world promote fair play values. Moreover, this project clearly showed the importance of program evaluation and the value of direct observation in a natural setting. PMID- 16248471 TI - Effects of creatine supplementation on aerobic power and cardiovascular structure and function. AB - This project aimed to determine 1) whether creatine (Cr) supplementation affects cardiovascular structure and function and 2) to examine its effect on aerobic power. Eighteen males undertook aerobic testing on a cycle ergometer and echocardiographic assessment of the heart. The experimental group (N = 9) ingested 20g x day(-1) of Cr for seven days followed by l0g x day(-1) for a further 21 days. The control group (N = 9) followed an identical protocol ingesting a placebo for the same period. Assessment was performed pre-, mid- (seven days) and post-testing (28 days). A MANOVA with repeated measures was used to test for group differences before and after supplementation. The Cr group demonstrated a significant increase in body mass for the pre-mid (1.0 +/- 0.6 kg) and the pre-post (1.5 +/- 0.7 kg) testing occasions. Submaximal VO2 decreased significantly from the pre-mid and pre-post testing occasions by between 4.8% to 11.4% with Cr supplementation at workloads of 75 W and 150 W. Other oxygen consumption measures and exercise time to exhaustion, for the Cr group, showed decreasing trends that approached significance. Additionally, there was a significant pre-post decrease in maximum heart rate of 3.7%. There were no changes in any of the echocardiographic or blood pressure measures for either group. The present results suggest short term Cr supplementation has no detectable negative effect on cardiac structure or function. Additionally, Cr ingestion improves submaximal cycling efficiency. These results suggest that the increase in efficiency may be related to peripheral factors such an increase in muscle phosphocreatine, rather than central changes. PMID- 16248472 TI - The detraining and retraining of an elite rower: a case study. AB - A heavyweight male rower, and current Olympic champion, undertook a laboratory based incremental rowing test on four separate occasions; eight weeks prior to the Sydney Olympics (Pre OG), after eight weeks of inactivity (Post-IA), after 8 weeks of retraining (Post 8) and after a further 12 weeks of training (Post 20). Following the period of inactivity, peak oxygen uptake (VO2peak) declined by 8%, power at reference blood lactate concentrations declined by approximately 100 W (25%), and power at VO2peak was 20% lower. With eight weeks of retraining, rapid improvements were seen. For most parameters, however, the rate of improvement slowed and after 20 weeks of retraining the individual was approaching pre Olympic levels. VO2 at lactate threshold as a percentage of VO2peak remained unchanged. These results show that detraining in the elite athlete can be pronounced, with rapid improvements upon retraining which slow, so that retraining takes considerably longer to achieve than detraining did. Complete cessation of training should be limited to short periods only in the preparation of the elite heavyweight rower. Any break should, if possible, include 'maintenance training'. In this way any decrements in those physiological parameters associated with 2000 m rowing performance will be minimised. PMID- 16248473 TI - Fitness testing and career progression in AFL football. AB - Relationships between fitness testing and career progression in the Australian Football League (AFL) are under-explored. This study investigated relationships between anthropometric and fitness tests conducted at the annual AFL National Draft Camp and subsequent career progression of players. A total of 283 players was tested over three consecutive camps (1999-2001). The anthropometric and fitness measures were: height, mass, sum of skinfolds, 20-m sprint test, vertical jump (standing and bilateral running), agility run and a multi-stage incremental shuttle run. The five outcome variables were: drafted (yes/no), AFL debut (yes/no), number of AFL games played to the end of 2003, and subjective ratings of career potential and career value (5-point scale). Of 205 players (72%) subsequently drafted, 166 (59%) eventually made their AFL debut. Players drafted to AFL clubs were faster over 5 m, 10 m and 20 m, ran further in the shuttle run and ran marginally faster in the agility test than players not drafted. Multi regression analysis showed small to moderate correlations (r = 0.27-0.31) between the designated outcome variables and selected fitness tests: 20-m sprint time (faster), agility run test (faster), and running vertical jump (higher absolute height and smaller difference between left and ride sides). Regression analysis for the standing vertical jump relative to standing reach height showed a counterintuitive negative correlation with positive outcomes, possibly reflecting non-compliance with testing procedures by the less successful athletes. We conclude that the 20-m sprint, jump, agility and shuttle run tests have a small but important association with career progression of AFL footballers. PMID- 16248475 TI - Development of a psychometrically valid and reliable sports nutrition knowledge questionnaire. AB - The present range of sports nutrition knowledge questionnaires have inadequate psychometric validation, and few are up to date in a rapidly changing discipline. The purpose of this study was to design a sports nutrition questionnaire that satisfied acceptable psychometric criteria of validity (content and construct) and reliability (test-retest). The questionnaire was designed by an expert panel of six sports dietitians and distributed to five groups, selected for their expected variation in sports nutrition knowledge. Dietitians, university business staff and nutrition students received questionnaires via e-mail. The response rates obtained were 21.3% (n = 49), 34.4% (n = 33), and 72.0% (n = 18), respectively. University business and fitness students completed questionnaires during class time. Response rates were 52.3% (n = 23) and 75.4% (n = 49), respectively. The questionnaire was administered a second time to the business staff and the dietitians to assess test-retest reliability. Two methods were used: 1, Pearson's product-moment correlation; and 2, a percentage calculation of questions answered in an identical manner on both test occasions. Reliability was acceptable with Method 1 yielding acceptable values (r = 0.74-0.93), aside from the fluid sub-category (r = 0.52). Method 2 showed good test-retest concordance with 81.2% duplication of responses of all questions. Construct validity was high, as indicated by significant mean knowledge score differences between the groups (p = 0.0001). Dietitians and nutrition students achieved significantly greater mean scores than the remaining groups. The findings of this study indicate that the questionnaire is suitably valid and reliable to be used in research and practice to determine sports nutrition knowledge. PMID- 16248474 TI - Physiological and anthropometric characteristics of starters and non-starters and playing positions in elite Australian Rules Football: a case study. AB - A purpose of this study was to determine if pre-season anthropometric and physiological measures were significantly different for the players from one Australian Football League (AFL) club selected to play in the first game of the season compared to the players not selected. Another purpose was to compare fitness test results for defenders, forwards and mid-fielders in the same AFL club. Thirty-four players were tested for isolated quadriceps and hamstrings strength, leg extensor muscle strength and power, upper body strength, sprinting speed, vertical jump (VJ), endurance, skinfolds and hamstring flexibility. The starters who were selected to play the first game were a significantly older and more experienced playing group, and were significantly better (p < 0.05) in measures of leg power, sprinting speed and the distance covered in the Yo Yo intermittent recovery test compared to the non-starters. Although there were trends for the superiority of the starters, the differences in lower and upper body strength, VJ and predicted VO2max were non-significant. The forwards generally produced the worst fitness scores of the playing positions with the midfielders having significantly lower skinfolds and the defenders possessing better hamstring strength and VJ compared to the forwards. It was concluded that some fitness qualities can differentiate between starters and non-starters, at least in one AFL club. Comparisons of playing positions and the development of fitness norms for AFL players require further research. PMID- 16248476 TI - Do squash players accurately report use of appropriate protective eyewear? AB - Self-report surveys are a common method of collecting data on protective equipment use in sport. The aim of this study was to assess the validity of self reported use of appropriate protective eyewear by squash players. Surveys of squash players' appropriate protective eyewear behaviours were conducted over two consecutive years (2002 and 2003) at randomly-selected squash venues in Melbourne, Australia. Over the two years, 1219 adult players were surveyed (response rate of 92%). Trained observers also recorded the actual on-court appropriate protective eyewear behaviours of all players during the survey sessions. Eyewear use rates calculated from both data sources were compared. The self-reported appropriate protective eyewear use rate (9.4%; 95% CI 7.8, 11.0) was significantly higher (1.6 times more) than the observed rate (5.9%; 95%CI 4.6, 7.2). This suggests that players may over-report their use of appropriate protective equipment, though some may have incorrectly classified their eyewear as being appropriate or suitably protective. Studies that rely only on self report data on protective equipment use need to take into account that this could lead to biased estimates. PMID- 16248477 TI - Defining a cricket injury. PMID- 16248478 TI - Separations in multiple-channel microchips. PMID- 16248480 TI - Lipophilicity measurements by liquid chromatography. PMID- 16248479 TI - Temperature effects in liquid chromatography. AB - Temperature has a large effect on retention, selectivity, and column efficiency and has long been accepted as an important parameter in liquid chromatography (LC). Despite this fact, temperature has not been very actively utilized in the past, mainly because of reported stability problems of the most commonly used stationary phases. However, more interest in the application of temperature for retention control has come of late because of the trend of miniaturization in chromatography and the availability of temperature-stable stationary phases. This work gives an overview of temperature effects on retention and selectivity in chromatography, especially on reversed-phase columns. Instrumental requirements, especially with respect to performing temperature gradient elution, are discussed and applications on both conventional-sized analytical columns and capillary columns are included. PMID- 16248481 TI - Concepts and practice of multidimensional high-performance liquid chromatography. PMID- 16248482 TI - High-performance liquid chromatography in the pharmaceutical industry: application, validation, and regulatory issues under the PAT framework. PMID- 16248483 TI - The use of polysaccharide phases in the separation of enantiomers. PMID- 16248484 TI - Chaotropic effects in RP-HPLC. PMID- 16248485 TI - Chromatography of difficult and water-insoluble proteins with organic solvents. PMID- 16248486 TI - Patterns of intimate partner violence: correlates and outcomes. AB - Battered women experience different constellations of violence and abusive behavior characterized by various combinations of physical violence, sexual violence, psychological abuse, and stalking. The goals of the current study were to determine whether it was possible to identify empirically derived and meaningful patterns of intimate partner violence (IPV) and to examine correlates and outcomes of the IPV patterns. Three IPV patterns were identified using cluster analysis. Pattern 1 was characterized by moderate levels of physical violence, psychological abuse, and stalking but little sexual violence. Pattern 2 was characterized by high levels of physical violence, psychological abuse, and stalking but low levels of sexual violence. Pattern 3 was characterized by high levels of all violence types. IPV Pattern 3 was associated with the highest prevalence of posttraumatic stress disorder and depression, and IPV Pattern 2 had the highest levels of revictimization during the year following recruitment. The clinical and policy implications of the findings are discussed. PMID- 16248487 TI - Longitudinal model predicting mutual partner violence among White, Black, and Hispanic couples in the United States general population. AB - This analysis determines the longitudinal predictors of male-to-female (MFPV) or female-to-male (FMPV) alone and mutual partner violence (MPV) among White, Black, and Hispanic couples. A national sample of couples 18 years of age or older was interviewed in 1995 and again in 2000. Participants constitute a multistage area probability sample representative of married and cohabiting couples from the 48 contiguous United States. Results indicate that most couples reporting violence engage in MPV. After controlling for other factors, Blacks are 3 times more likely to report MPV at follow-up and Hispanics are 9 times more likely to report MFPV. The results indicate that ethnic minorities are at greater risk of MPV. In addition, the predictors of partner violence vary depending on the type of partner violence. These findings highlight the importance of distinguishing different types of partner violence and have important epidemiological and prevention implications. PMID- 16248488 TI - Exploring the relation between bullying and homophobic verbal content: the homophobic content agent target (HCAT) scale. AB - This investigation quantitatively examines the association among homophobic content, bullying, victimization, empathy, and several psychosocial outcomes of these constructs. The 2-factor Homophobic Content Agent Target (HCAT) scale was developed and validated among 191 middle school students to assess the extent to which students both use and are called various epithets in reference to sexual orientation. Cronbach reliability coefficients of alpha = .85 were obtained for both factors. Convergent validity was demonstrated with scales measuring bullying, fighting, victimization, relational aggression and victimization, anxiety and depression, and delinquency. Discriminant validity was demonstrated in comparison with school sense of belonging, empathy, and perspective-taking. Discriminative validity was demonstrated through sex differences on several scales. Results strongly suggest that homophobic content is prevalent in various forms of aggression and victimization, and that future research should examine the role of homophobia in bullying and victimization in schools. PMID- 16248489 TI - The composite abuse scale: further development and assessment of reliability and validity of a multidimensional partner abuse measure in clinical settings. AB - Absence of a well-validated comprehensive partner abuse questionnaire has been a major methodological issue in domestic violence research. A new multidimensional measure of partner abuse, the Composite Abuse Scale (CAS), has four dimensions: Severe Combined Abuse, Emotional Abuse, Physical Abuse, and Harassment. A general practice patient sample (N = 1,836) has been used in the development and testing of CAS. Factor analyses in this current study confirmed the four dimensions from a preliminary nurses sample study and resulted in a final scale of 30 items consisting of acts of physical, emotional, and sexual abuse. These four factors exhibited good internal reliability (Cronbach's alpha > 0.85) and the corrected item-total correlations were high (> 0.5). Evidence of criterion and construct validity is presented. PMID- 16248490 TI - Gender, partner violence, and perceived family functioning among a sample of Vietnam veterans. AB - This study examined partner violence and perceived family functioning among a sample of 298 male veterans and their female partners. Partner violent men were higher than partner violent women on measures of partner violence severity, although differences did not reach statistical significance. Among couples experiencing unidirectional violence, female victims of partner violence reported significantly poorer family functioning than male victims of partner violence. Data appear to suggest that the effects of male-perpetrated partner violence on perceived family functioning may be larger than that of female-perpetrated partner violence. PMID- 16248491 TI - Prevalence of victims of violence among ethnically diverse Asian/Pacific Islanders. AB - The present research project is the first large-scale study (N = 5,051) that investigated the prevalence of victims of violence for an ethnically diverse Asian/Pacific Islander sample. The rate for the adolescent respondents of "was a victim of violence (was physically harmed by someone)" within the past 6 months was 3.33%. Over twice that rate was found for family members (6.97%) and over three times the adolescent-respondent rate was obtained for close friends (10.75%). Only partial support was found for the hypothesis that Asian groups would have the lowest rates, and Polynesian, African American, Hispanic, and Native American Indian/Alaska Native groups would have the highest rates. Higher rates were found for Whites (adolescent respondents, close friends) and the Portuguese (family members, close friends), suggesting a "minority" effect. There is a need to disaggregate ethnicity, engage in further research that considers risk and protective factors, and investigate culturally appropriate interventions. PMID- 16248492 TI - College women's experiences of psychological maltreatment and sexual assault. AB - A sample of 648 college women recruited from undergraduate psychology classes was examined to explore the relationship between past psychological maltreatment and sexual assault. Based on responses to the Sexual Experiences Survey and the Psychological Maltreatment Inventory, women were classified by level of unwanted sexual contact (i.e., vaginal or anal intercourse; oral genital contact and/or object penetration; or kissing and/or fondling), by method used to obtain the sexual assault (i.e., women were classified as experiencing coerced assaults, forced assaults, or both), and by identity of perpetrator (i.e., acquaintances or strangers). Results pointed to significant differences in the amount of past psychological maltreatment reported by women experiencing any type of assault as compared to women without assault experiences, regardless of perpetrator identity. Moreover, higher levels of psychological maltreatment were associated with having experienced any type of coerced sexual activities. There were no differences by type of assault. Finally, a series of ANOVAs was conducted to examine the interaction between coercion and force in the psychological maltreatment reported by women experiencing different forms of assault. With few exceptions, greater maltreatment was associated both with the occurrence of coerced assaults and with the occurrence of forced assaults. A significant interaction was seen with one form of assault, unwanted kissing and/or fondling perpetrated by an acquaintance. This interaction may suggest that, at least for this one form of contact perpetrated by acquaintances, the presence of past psychological maltreatment is uniquely associated with experiencing adult sexual assaults involving both force and coercion. PMID- 16248493 TI - Effects of trauma exposure on anger, aggression, and violence in a nonclinical sample of men. AB - This study assessed the impact of traumatic exposure and posttraumatic stress disorder (PTSD) symptoms on anger, aggression, and violence among civilian male college students. Results suggest that civilian men who have been exposed to a potentially traumatic event (PTE) and report symptoms of PTSD indicate more trait anger, more internal anger and hostility, and more aggression and violence than men who do not report symptoms of PTSD. Results are contrasted to those found in clinical samples of male veterans with PTSD and discussed in terms of understanding and treating anger and aggression in non-clinical, trauma-exposed populations. PMID- 16248494 TI - German capitalization of nouns and the detection of letters in continuous text. AB - The missing-letter effect refers to the phenomenon that letters are more difficult to detect in common function words (such as the) than in content words. Assuming that the missing-letter effect is diagnostic of the extraction of text structure, we exploited a special feature of German--the convention to capitalize the initial letter of nouns. Given the great flexibility of word order in German, it was proposed that this convention might help readers specify the structure of the sentence. Therefore orthographic variations that violate the capitalization rules should disrupt structure extraction and should result in a reduced missing letter effect. The results indicated that: 1) capitalization of function words eliminated the missing-letter effect, but not at the beginning of a sentence; 2) A missing-letter effect occurred when the capitalization of the first letter was correct, but was followed by typecase alternation, and also when the size of the initial letters was relatively large for function words, but relatively small for content words. The results were discussed with respect to the possible contributions of visual familiarity, structural role, and processing time to the missing-letter effect, taking into account that a capitalized initial letter conveys significant information about the word class for German readers. Thus, the present results indicate that readers take advantage not only of function words but of any other information (here the capitalization of nouns) that helps to extract the structure of a sentence. PMID- 16248495 TI - Trade-offs in detecting letters and comprehending text. AB - Critics of the letter detection task have questioned whether findings from that paradigm reflect normal reading processes. The present study addresses these questions using a new computerized version of the letter detection task in which reading rate along with letter detection and comprehension accuracy are examined. Previous letter detection findings were replicated with this new computerized task. Different conditions were compared in which detection and comprehension instructions were manipulated as well as the salience of the target letters. The requirement to comprehend had small effects on letter detection accuracy and reading rate, and letter detection only modestly reduced comprehension. Thus, the procedures developed in this study permit examination of the component processes contributing to performance in the letter detection task. PMID- 16248496 TI - Reasoning about conjunctive probabilistic concepts in childhood. AB - While adults are known to exhibit biases when making conjunctive probability judgments, little is known about childhood competencies in this area. Participants (aged between four and five years, eight and ten years, and a group of young adults) attempted to select the more likely of two events, a single event, and a conjunctive event containing, as one of its components, the single event. The problems were such that the objective probabilities of the component events were potentially available. Children in both age groups were generally successful when the single event was likely. However, when it was unlikely, a majority of children rejected it, choosing the conjunctive event instead, thereby committing the conjunction fallacy. A substantial minority of adults also committed the fallacy under equivalent conditions. It is concluded that under certain conditions children are capable of normative conjunctive judgments but that the mechanisms underpinning this capacity remain to be fully understood. PMID- 16248497 TI - [Judgement of Authenticity of Smiles and Detection of Facial indexes]. AB - The smile is one of the most often expressed emotions during social interactions. It can be authentic, that is, associated with a joyful emotional state in the person expressing it, but it can also be false, that is, deliberately produced in the absence of that emotional state in order to deceive one or more individuals (Ekman, 1993). Even though the fake smile very much resembles the authentic smile, it generally does not constitute the perfect simile. The fake smile more often has a certain degree of asymmetry than the authentic smile (Ekman, Hager, & Friesen, 1981) and it uses the cheek raiser action less often than with the authentic smile (Ekman, Friesen, & O'Sullivan, 1988; Frank, Ekman, & Friesen, 1993). This study looked at the knowledge that adults have of these differences as well as their perceptive ability to detect them. The visual stimuli presented to participants were prepared using the Facial Action Coding System (Ekman & Friesen, 1978). Results show that participants detected the differences between the two types of smile and that detection was better using smile asymmetry than with the cheek raiser action. Analysis of the use of response categories in the detection task indicated that participants underestimated the differences between smiles when they were different and that this tendency was more apparent with the cheek raiser detection method than for asymmetry detection. Participants also demonstrated a better knowledge of smile asymmetry than cheek raiser action. The knowledge gathered suggests that the ability of the receptor to judge smile authenticity is limited by perceptive factors. However, the mediation analyses that we conducted show the judging smile authenticity is not limited to simple perceptive detection of facial clues. Detecting facial clues is a necessary condition for correctly assessing smile authenticity, but it does not explain the variance in these assessments. We believe that this variance would be due more to the importance that participants give to facial clues. Finally, our results show that the capacity to detect differences between authentic and fake smiles is not easy to change. Participants who received modified information on changes of appearance linked to the two facial parameters were not more likely to detect the differences than participants who did not receive information. PMID- 16248498 TI - [Distribution effects of orthographic similarity and priming by masked repetition]. AB - This research centres on the effect that the orthographic neighbourhood has in the visual recognition of words. Specifically, we studied to what extent orthographic neighbourhood distribution, that is, the number of letter positions allowing formation of at least one neighbour (Pugh, Rexer, Peter, & Katz, 1994), influences the masked repetition priming effect. In a previous study (Mathey, Robert, & Zagar, 2004), interaction between neighbourhood distribution and orthographic priming was obtained in the lexical decision task. The Interactive Activation Model (IA; McClelland & Rumelhart, 1981) simulated this interaction. With the orthographic priming effect modified for distribution of the neighbourhood of target words, it was necessary to study whether the repetition priming effect also varied as a function of this indicator. Studying this interaction presents a major theoretical issue in specifying the activating and inhibiting processes presented in the IA model. Simulations were produced to obtain precise model predictions regarding the neighbourhood distribution effect in a repetitive priming situation for our experimental material. Target words all had two neighbours that were most frequent. These neighbours were isolated, that is, distributed over two letter positions (e.g.: TAUX/faux-toux), or associated, i.e., concentrated on one single position (e.g., SEAU/beau-peau). Targets were preceded by an identical priming (repetitive priming; e.g.: seau-SEAU) or by controlled priming (e.g., &&&&-SEAU). The simulation results obtained using the IA model show the facilitating effects of neighbourhood distribution and repetitive priming, but no interaction between these factors. The experimental results obtained in a lexical decision task confirm these predictions. Thus, the empirical data replicate the neighbourhood distribution's facilitating effect (Mathey & Zagar, 2000) as well as the facilitating effect of masked repetition (Forster & Davis, 1984). Finally, the most interesting result is that the facilitating effect of repetition is comparable for target words with associated neighbours and target words with isolated neighbours. An explanation of the combined effects of the orthographic neighbourhood and orthographic masked repetition priming, integrating data from literature as well as from the current study, is proposed within the framework of the IA model. PMID- 16248499 TI - Making sense of memory. AB - The current work explores how people make recognition and belief judgments in the presence of obvious repetition primes. In two experiments, subjects received a 200-ms prime ("cheetah"), either before or after reading a trivia question ("What is the fastest animal?") but always before being presented with the target answer ("cheetah"). Results showed that repetition priming decreased "old" claims (Recognition--Experiment 1), while it increased truth claims (Belief--Experiment 2). Furthermore, repetition prime placement affected recognition but not belief. Combined, these results suggest that dissociations in memory performance are a natural outcome of task and processing demands and reflect the dynamic, flexible nature of memory. PMID- 16248500 TI - Eye movements and phonological parafoveal preview: effects of reading skill. AB - Eye movements of skilled and less skilled readers were monitored as they read sentences containing a target word. The boundary paradigm was used such that when their eyes crossed an invisible boundary location, a preview word changed to the target word. The preview could either be identical to the target word (beach as a preview for beach), a homophone of the target word (beech as a preview for beach), an orthographic control (bench as a preview for beach), or an unrelated consonant string (jfzrp as a preview for beach). Consistent with prior research, skilled readers obtained more preview benefit from the homophone preview than from the orthographic preview. The less skilled readers, however, did not show such an effect. The results indicate that less skilled readers do not use phonological codes to integrate information across eye movements. Indeed, the results also indicate that less skilled readers do not show normal preview benefit effects. PMID- 16248501 TI - [Genioglossus advancement and hyoid suspension plus uvulopalatopharyngoplasty for severe OSAHS]. AB - OBJECTIVE: To explore a comprehensive surgical approach of genioglossus advancement and hyoid suspension plus uvulopalatopharyngoplasty on the treatment of severe obstructive sleep apnea hypopnea syndrome. METHOD: Eighteen patients with severe OSAHS (RDI > 40)diagnosed by polysomnography. The obstruction in both the oropharynx and the hypopharynx were determined by preoperative physical examination, fiberoptic pharyngolaryngoscopy, cephalometry, and computed tomography of the upper airway. All the cases were given continuous airway pressure via nose,five to seven days before operation and then were performed genioglossus advancement and hyoid suspension plus uvulopalatopharyngoplasty. The follow up was at least 6 months postoperatively. The Wilcoxon Signed Rank test was used to compare the preoperative and postoperative results by SPSS11.0 for windows. RESULT: Postoperative 6-24 months, there were statistically significance in all but BMI between preoperative and postoperative measurement. Mean RDI was reduced from preoperative (63.83 +/- 16.34) to postoperative (21.43 +/- 20. 34), lowest mean oxygen saturation increased from (72.44 +/- 7.07)% to (81.33 +/- 13.32)%. According to criterion at home, the 6-month rate of responder is 83.33%. The severe complications such as fracture of the mandible and injure of apex of inferior anterior teeth did not occurred. CONCLUSION: GAHM plus UPPP is effective surgical approach for patients with severe OSAHS who suffer from oropharyngeal and hypopharyngeal obstruction. PMID- 16248502 TI - [The features of auditory brainstem response in patients with obstructive sleep apnea-hypopnea syndrome]. AB - OBJECTIVE: To investigate the effects and causes of obstructive sleep apnea hypopnea syndrome (OSAHS) on auditory pathway, the changes of auditory brainstem response (ABR) were analyzed. METHOD: With the use of polysomnography,72 snorers were examined and divided into OSAHS and non- OSAHS groups. These two groups patients and a group of normal person were underwent ABR measuring. RESULT: There were statistically significant difference between the group of OSAHS and the non OSAHS or the group of OSAHS and the normals on the delayed latency of wave I and wave V, the response liminal value of wave V rised. CONCLUSION: The results suggest that cochlear function may be affected in patient with OSAHS, the effected of brain stem is unknown. PMID- 16248503 TI - [The effect of sodium phenylbutyrate to agents used in induction chemotherapy on laryngeal carcinoma cells Hep-2 in vitro]. AB - OBJECTIVE: To study the effect of sodium phenylbutyrate when it combined with agents used in induction chemotherapy on laryngeal carcinoma cells Hep-2 in vitro. METHOD: MTT were used to examine the growth inhibition of Hep-2 cells treated by the combination of PB with 5-FU or CDDP in vitro. RESULT: When 5-FU or CDDP combined with PB respectively, there was significantly difference between every two dose groups of the two agents or every dose group and control group ( P < 0.05). When the dosage of 5-FU or CDDP was definition,there was significantly difference between every two dose groups of PB ( P < 0.05). CONCLUSION: PB could enhance the cytotoxic effects of agents used in induction chemotherapy on laryngeal carcinoma cells Hep-2 in vitro, which showed the possibility in reinforcement the treatment effect and reduction the occurrence of the complication and toxic reaction of induction chemotherapy on laryngeal carcinoma. PMID- 16248504 TI - [Investigate the early impairment of renal function caused by chronic tonsillitis]. AB - OBJECTIVE: To investigate whether the chronic tonsillitis have early impairment of renal function or not. METHOD: Enzyme linked immunosorbent assay (ELISA) was used to detect the urinary microalbumin of 77 samples of chronic tonsillitis (group A: course of diseases is 1 to 3 years; group B: course of diseases is 3 to 5 years; group C: course of diseases is over 5 years) and 20 samples of normal (group D) respectively and the results were compared, including the detection and comparison of the urinary microalbumin of 19 samples of chronic tonsillitis preoperative and postoperative. RESULT: (1) group A: the urinary microalbumin is (9.8 +/- 1.6)mg/L; group B: the urinary microalbumin is (12.3 2.2) mg/L; group C: the urinary microalbumin is (17.6 +/- 3.2)mg/L; group D: the urinary microalbumin is (9.6 +/- 2.0) mg/L. The urinary microalbumin of chronic tonsillitis group C was higher than group D, there was a significant difference between these two groups ( P < 0.05). (2) The urinary microalbumin of chronic tonsillitis was (26.3 +/- 2.2)mg/L preoperatively and was (18.7 +/- 1.7)mg/L postoperatively. The urinary microalbumin of chronic tonsillitis significantly decreased after they had been surgical treated (P < 0.05). CONCLUSION: The urinary microalbumin of chronic tonsillitis was uptrend according to its course of diseases. The detection of the urinary microalbumin could be one of the diagnostic message for the early impairment of renal function which has been caused by chronic tonsillitis. The detection of the urinary microalbumin is important to evaluate the surgical treatment of chronic tonsillitis. PMID- 16248505 TI - [The study of phonatory function in myasthenia gravis patients]. AB - OBJECTIVE: To investigate the phonatory function in myasthenia gravis patients and the role of acoustic analysis technique in the diagnosis and the assessment of therapeutic effect on treatment of myasthenia gravis. METHOD: Eighty-three myasthenia gravis patients diagnosed definitely in neurology were divided into two groups, ocular group and generalized group. Each group included male group and female group. Acoustic measures were carried out before and after neostigmine test and treatment. Forty healthy subjects including twenty male and twenty female served as control groups. The basic parameters of the voice using computed voice analysis included F0, shimmer, jitter and NNE. Independent-Samples T Test and Paired-Samples T Test were used. RESULT: The value of F0 and NNE increased in generalized group. The changes of F0 and NNE in generalized group were found to be statistically significant (P < 0.01, 0.05). Neostigmine resulted in an improvement in the parameters of F0 and NNE, the same condition happened after the treatment (P < 0.01). CONCLUSION: Phonatory function of generalized myasthenia gravis patients is abnormal. F0 and NNE increase in generalized myasthenia gravis patients and decrease after the treatment or the intake of neostigmine. Acoustic analysis is a useful method in the diagnosis and assessment of generalized myasthenia gravis patients. PMID- 16248506 TI - [Expression and significance of CD44v6 and MMP-9 in laryngeal squamous cell carcinoma]. AB - OBJECTIVE: To investigate the clinical implication and the expression of CD44v6 and MMP-9 protein in laryngeal squamous cell carcinoma (LSCC). METHOD: Twenty seven samples of LSCC and eight biopsies of adjacent non-cancerous tissue were detected with immunohistochemical technique (SABC method). The relationship between positive expression of CD44v6 or MMP-9 in LSCC and invasion or metastasis was analyzed. RESULT: Both CD44v6 and MMP-9 overexpressed in LSCC. The positive expression of CD44v6 was correlated with LSCC pathologic grade, cervical lymph node metastasis and postoperation survival rate (P < 0.05). The expression of MMP 9 was positively correlated with clinical staging and cervical lymph node metastasis (P < 0.05). There was a significant positive correlation between the expression of CD44v6 and MMP-9 (P < 0.01). CONCLUSION: There is a relationship between the expression of CD44 and MMP-9. CD44v6 and MMP-9 might be served as an objective indicator for lymph nodal metastatic potency, tumor invasion and prognosis in LSCC. PMID- 16248507 TI - [Application of partial laryngectomy in laryngeal cancer treatment and reconstruction with epiglottic flap and bi-pedicle muscle flap]. AB - OBJECTIVE: To explore and evaluate the surgical method of partial laryngectomy and the laryngeal defects were reconstructed. METHOD: Two hundreds and sixty-five patients with laryngeal cancer were treated by partial laryngectomy and the laryngeal defects were reconstructed by epiglottic flap, bi-pedicle sternohyoid muscle fascia flap and epiglottic flap plus bi-pedicle sternohyoid muscle fascia flap from January 1990 to December 2000. There were 256 males and 9 females,ranging in age from 48 to 75 years old. Neck dissection were performed in 79 patients. RESULT: The total cases began to eat between 10 to 15 days after operation. Two hundreds and eighteen cases were decannulated with a decannulation rate of 82.26%. Wound infection occurred in 12 cases and 2 pharyngocutaneous fistula was found. The three and five years survival rates were 74.72% and 70. 88% respectively. CONCLUSION: After partial laryngectomy in laryngeal cancer epiglottic flap and bi-pedicle muscle fascia flap were performed reconstruction of laryngeal function in a single-stage. These procedures which were relative simplicity, expediency, small injury and high rate of survival present ideal effects and are worthy to be recommended. PMID- 16248508 TI - [The detection of human papilloma virus in juvenile-onset respiratory papillomatosis]. AB - OBJECTIVE: To investigate the relationship between juvenile-onset respiratory papillomatosis (JOP) and human papilloma virus (HPV6, HPV11), and the immune function of patients. METHOD: Fluorescence quantitative PCR (FQ-PCR) which combines PCR and fluorescence probe hybridization was used to detect HPV6 , HPV11 DNA in 130 cases. Of these, 68 cases were used Flow Cytometry to measure CD3, CD4, CD8. RESULT: One hundred and fifteen of 130 cases were HPV6, HPV11 DNA positive, the average copy was 5.68 +/- 2.65. For 68 cases,the average percent of CD3,CD4,CD8 were 62.73 +/- 8.63, 30.54 +/- 7.05, 26.08 +/- 6.93, respectively. To compare with control group, there was not statistical significance. CONCLUSION: FQ-PCR is a convenient, accurate and specific method which can quantify the amount of pathogenic germs from 10(1) to 10(10), and is also a reliable factor to predicate clinical diagnose and cure. PMID- 16248509 TI - [To treat pharyngeal bursitis by microwave under nasal endoscope]. AB - OBJECTIVE: Discussing the curative effect to treat pharyngeal bursitis by microwave under nasal endoscope, observing the wound change and writing down the time of white scar dropping after the operation. METHOD: Eleven cases of pharyngeal bursitis and 6 cases of nasopharyngeal cyst were treated by microwave under nasal endoscope, 0.9% sodium chloride solution were given for nasal irrigation two times daily,to observe the wound under nasal endoscope weekly up to the white scar dropping. RESULT: There are 14 cases cure and 3 cases in effect. The white scar dropped between 2 to 3 weeks and the patients symptom disappeared. CONCLUSION: Nasal endoscopy conduce to the diagnosis of pharyngeal bursitis. It is an effective method to treat pharyngeal bursitis by microwave under nasal endoscope and 0.9% sodium chloride solution were given for nasal irrigation after operation. PMID- 16248510 TI - [The treatment of nasal adhesion after endoscopic sinus surgery]. AB - OBJECTIVE: To explore the cause and the handle method of nasal adhesion after endoscopic sinus surgery (ESS). METHOD: There are 224 patients' clinical datum after ESS. Twenty-one patients were chosen after the operation of nasal adhesion. Analysis this group of the medical record and comparing with that 203 patients who haven't adhesion. RESULT: Among the 21 patients who have been adhesive, there are 15 patients who have been adhesive in middle turbinate and lateral nasal wall, 6 patients who have been adhesive in nasal septum and inferior turbinate. CONCLUSION: There are three main chief causes of adhesion, short distance in the operation's trauma, dressing changing not in time after the operation and mistaken method after nasal adhesion. PMID- 16248511 TI - [Experimental study of tracheal defect repair with shape-memory titanium-nickel alloy net]. AB - OBJECTIVE: To investigate the feasibility of trachea defect repair with "C" shape shape-memory titanium-nickel alloy net (CSSMAN). METHOD: The CSSMAN substitute for the tracheal cartilage and the skin tube for the tracheal mucosa in order to form a new "trachea" shape structure to repair the mongrel dogs trachea defect. RESULT: Four dogs were successfully decannulate, computerized tomography (CT) examination and fibrotracheoscopy examination showed the CSSMAN located perfectly and the airway is wide enough for breath. One dog failed to decannulate due to the infection. CONCLUSION: It is a promising choice to repair the trachea defect with CSSMAN by two-stage procedure. PMID- 16248512 TI - [Antitumor research on human laryngeal carcinoma with combined application of VP3 gene, IL-18 gene and HN gene]. AB - OBJECTIVE: To research the antitumor efficacy on mice model bearing H22 tumor with application of pVVP3IL-18HN and its mechanism of the antitumor effect on Hep 2. METHOD: pVVP3IL-18HN was introduced into Hep-2 cells by liposome, then cellular morphology was observed through AO/EB stain. To evaluate the mechanism of the antitumor effect on Hep-2 of pVVP3IL-18HN and its effect on tumor immunogenicity, flow cytometer (FCM) was used to detect the variation of mitochondrial trans-membrane potential,reactive oxygen species (ROS) and the MHC I. C57BL/6 mice were subcutaneously inoculated with 2 X 10(5) H22 tumor cells in the right hindlimb, and 7, 14 and 21 day post-inoculation, pVVP3IL-18HN was administrated intratumorally. The pVVP3, pVIL-18, PVHN and PBS control group were treated as above. Then the tumor suppression and specific cytotoxic T lymphocyte (CTL) activity was detected. RESULT: After the transfection of pVVP3IL-18HN, tumor cells crenated and stained by AO/EB. In the research pVVP3IL-18HN was found to up-regulate the value of ROS and the expression of MHC-I, and down-regulate the value of mitochondrial trans-membrane potential. The tumor suppression rate of pVVP3IL-18HN was 60.5% and a dramatically increased tumor suppression was revealed in the pVVP3IL-18HN treated mice as compared with control mice treated with PBS (P < 0.05). CONCLUSION: pVVP3IL-18HN kill the tumor cells by apoptosis and have notable antitumor effect in vivo. PMID- 16248513 TI - [The diagnoses and classifying of congenital craniofacial cleft]. AB - OBJECTIVE: To diagnose and classify the congenital craniofacial cleft with a uniform scale is helpful to evaluate the abnormality and select the repairing methods. METHODS: We analyzed 81 cases of congenital craniofacial cleft basically using Tessier craniofacial cleft classification. Furthermore, according to the position of soft tissue or bone, the character and degree of clefts or dysplasia and the results of CT scanning, we subdivided the congenital deformities based on S (skin), T (tissue), and O (OS). Arabic numerals were used to express the degree of the abnormality. RESULTS: Of all the cases analyzed with the STO classification, No. III and IV clefts are often seen in the infraorbital region (24.70%). No. IX and X clefts are mostly seen in the supraorbital region (38.27%). The relationship between the cleft types and involved tissue has not been found. CONCLUSIONS: The STO classification reinforces Tessier classification. It offers the basis for craniofacial cleft repair. PMID- 16248514 TI - [Mandibular distraction osteogenesis in the treatment of obstructive sleep apnea syndrome in children with micrognathia]. AB - OBJECTIVE: To evaluate the treatment of obstructive sleep apnea syndrome (OSAS) in children with congenital micrognathia using mandibular distraction osteogenesis and to discuss the advantages and disadvantages of this approach. METHODS: 6 patients (4 males, 2 females) had undergone mandibular distraction osteogenesis, 12 distraction devices were placed through extraoral incision for bilateral distraction. The mean age of treatment was 1 year and 9 months (range 4 months to 9 years). Every patient had been evaluated pre and postoperatively with cephalometry. The period of consolidation was 4 - 11 weeks. The period of follow up was from 2 to 10 months. RESULTS: The average distraction distance was 19.2 mm (range 15 to 25 mm). The osteotomy and distraction processes were smooth in all the cases, the osteogenesis was good, without infection and other complications. The posterior airway space was increased from averaged 4.5 mm preoperatively to 10. 1mm after surgery. Five children had normal respiration and sleep restored with naso-pharyngeal airway removal or tracheostomy decannulation. One patient is to receive a planned second stage of distraction with a horizontal vector. The results were stable without relapse during a follow-up period of 2 to 10 months. CONCLUSIONS: Application of mandibular distraction osteogenesis is an important component and effective in the treatment of OSAS and permits mandibular advancement in the younger child. As more experience is gained with distraction osteogenesis in the treatment of children with OSAS, the role of distraction will become better defined. PMID- 16248515 TI - [Clinical applications of devitalized autogenous calvarial bone flaps fixed with titanium clamps (CranioFix) for cranioplasty]. AB - OBJECTIVE: To evaluate reimplantation of devitalized autogenous calvarial bone flaps and the fixation of titanium clamps (CranioFix) for cranioplasty. METHODS: From March 2000 to December 2003, 21 patients underwent (1) delayed reimplantation of preserved autogenous bone flaps, which were removed during the initial operations for increased intracranial pressure and stored in a freezer and autoclaved before use, or (2) immediate reimplantation of tumor invaded bone flaps that were devitalized by boiling. All of these flaps were fixed with titanium clamps (CranioFix). Resorption of the bone flaps was monitored by skull roentgenograms and computed tomography (CT). The clinical and aesthetic results were evaluated. RESULTS: During the follow-up period of 6 to 30 months, no complications were found. There was slight resorption of the bone flaps in all cases, but the edge of the bone flaps was within the range of the titanium clamps according to skull roentgenograms and CT. Clinical and aesthetic results were highly satisfactory. CONCLUSIONS: It is simple and practical using devitalized autogenous calvarial bone flaps for cranioplasty. The firmness decreasing of the reimplanted bone flap due to resorption can be prevented greatly by titanium clamps (CranioFix). PMID- 16248516 TI - [Complications of scalp coronal incision: analysis, prevention, and treatment]. AB - OBJECTIVE: The aim of this study was to analyze the complications of coronal incision and investigate the methods of prevention. METHODS: The retrospective analysis was based on 149 cases, who have had operations since 1997 for congenital craniofacial malformation, second deformation of craniomaxillofacial trauma, maxillofacial tumor or cosmetic purposes. RESULTS: Of them, there were injury of unilateral frontal branch of the facial nerve in 3 cases, subcutaneous hematoma in 9 cases, alopecia in 12 cases, incision scar in 14 cases, obvious strip scar in 2 cases, pains, numbness and paraesthesia in 23 cases, ptosis of facial soft tissue in 8 cases and infection in 4 cases. CONCLUSIONS: The coronal incision has the merits of distinct exposure, hidden incision scar, but its complications can not be neglected. During the operation, care should be taken to anatomical layers, protecting the blood vessel and nerve bundle in order to reduce complications. PMID- 16248517 TI - [The clinic study of internal-fixation of mandibular fracture with endoscope]. AB - OBJECTIVE: To investigate effectiveness of internal-fixation in mandibular fracture through endoscope. METHODS: Use puncture internal-fixation under endoscope to treat 28 patients of fracture of mandibular angle, ramus and condyle. RESULTS: The 28 patients were restorated with occluding relation in good apposition. The healing of mandibular fracture was very good. CONCLUSIONS: The interal-fixation under endoscope is an effective way of treating fracture of mandibular angle, ramus and condyle, without facial scar and complication of facial nerve injury. PMID- 16248518 TI - [Classification and repairing investigation of the whistling deformities in the patients with bilateral cleft lip or bilateral cleft lip and cleft palate after primary cleft lip repair]. AB - OBJECTIVE: To classify the whistling deformities of the patients with bilateral cleft lip or bilateral cleft lip and cleft palate after primary cleft lip repair. METHODS: According to exposure degrees of the gum and the crown of the anterior upper incisors, 136 patients with whistling deformities after primary repair of bilateral cleft lip at the Department of Oral and Maxillofacial Surgery of Xiang Ya Hospital, Central South University were examined and the whistling deformities were classified into four grades. Grade I: the crowns of the left and the right upper incisors were exposed about one-second. Grade II: the whole crowns of the upper incisors were exposed or plus the exposure of the crowns of medial sagittal portion of the left and the right upper lateral incisors. Grade III: the whole crowns of two upper incisors and one-second of the attached gum of two upper incisors or the crowns and one-second of attached gum of the two upper incisors with over two-third of transverse crowns of the lateral incisors were exposed. Grade IV: the crowns and corresponding attached gum of the upper incisors and over two-third of medial sagittal crowns of the upper lateral incisors were exposed. The relation between the degrees of the whistling deformities and the primary procedures of the bilateral cleft lip or the bilateral cleft lip and cleft palate was analyzed. RESULTS: Sixty patients (44.2 percent) had grade I whistling deformities; forty-seven patients (34.5 percent) were Grade II; 16 patients (11.8 percent) were Grade III; 13 cases (9.5 percent) were Grade IV. The results showed that grade I and grade II whistling deformities were obviously more (about 3.7 times) than grade IlI and grade IV whistling deformities. CONCLUSIONS: Classification of whistling deformities provided the objective basis for the selection of operation techniques of cleft lip. PMID- 16248519 TI - [Repair of upper lip defects with an island flap based on the inferior labial arterial arch]. AB - OBJECTIVE: To investigate one-stage repairing upper lip defects with an island flap based on the inferior labial arterial arch. METHODS: An island flap pedicled with the inferior labial arterial arch was used to repair upper lip defects. The size of the wedge-shaped flap ranged from 1.5 cm x 1.5 cm to 1.5 cm x 2.0 cm. Since 1993, fifteen patients with upper lip defects have been treated with this method. RESULTS: All the flaps survived with satisfactory function and appearance. CONCLUSIONS: The island flap of the inferior labial arterial arch is effective and applicable in repairing severe secondary deformities of congenital bilateral cleft lips. PMID- 16248521 TI - [The reverse flow island flap nourished by sural nerve nutrition blood vessel for repair of severe frostbite of feet]. AB - OBJECTIVE: The study was to verify the feasibility of an improved method using reverse flow island flap nourished by the sural nerve nutrition vessel to repair severe frostbite of feet. METHODS: At the proximal end of the principal flap, an auxiliary triangular skin flap of 6.5 - 7 cm in length was designed in order to cover the pedicle of the principal flap. This operation was performed on 13 patients (21 feet) with frostbite. RESULTS: All the flaps survived well. Postoperative follow-up for 5 - 18 months demonstrated satisfactory results in all the cases. No ulcer happened. CONCLUSIONS: The method is helpful to prevent constriction of the pedicle and ensure blood supply of the flap. It is an ideal treatment for severe frostbite of feet. PMID- 16248520 TI - [The reverse buccinator musculomucosal flap for the treatment of inverted papilloma of the nose]. AB - OBJECTIVE: This paper presents the experience in using the reverse buccinator musculomucosal flap for repairing the defect following excising inverted papilloma of the nose. METHODS: After the inverted papilloma of the nose was excised through an endonasal approach, the reverse buccinator musculomucosal flap supplied by the retrograde blood flow of the anterior buccal artery was harvested and sutured to the defect through the ora-nasal tunnel. The procedure was performed on three patients. RESULTS: The postoperative course was uneventful. All the flaps survived completely. CONCLUSIONS: The technique provides the solution to prevent nasal stricture from cicatricial contracture after excising inverted papilloma. In the operation, excising the inverted papilloma and repairing the defect was performed simultaneously, saving another operation for the secondary deformity. The technique is also applicable to the treatment of existing cicatricial stricture of the nose. PMID- 16248522 TI - [Combination of multiple procedures to repair of severe lower eyelid ectropion]. AB - OBJECTIVE: Lower eyelid ectropion is one of the most severe complications following lower eyelid blepharoplasty. Combination of multiple procedures was used to repair this deformity and the results were observed. METHODS: Severe lower eyelid ectropion following blepharoplasty was repaired with the combination method of multiple procedures, including transferring the island forehead flap, buried guiding suture of the lower eyelid skin to orbital periosteum and horizontal lid-shortening. 10 patients (15 eyelids) with lower eyelid ectropion after blepharoplasty were treated with the above method from Mar 2001 to Mar 2004. RESULTS: The lower eyelid ectropion was repaired effectively. All the patients were satisfied with the results. CONCLUSIONS: The combination method of multiple procedures was an effective method for severe lower eyelid ectropion. PMID- 16248523 TI - [Applied anatomic investigation of phalloplasty ]. AB - OBJECTIVE: To investigate the anatomy of penis and its adjacent organ for phalloplasty. METHODS: Anatomic dissection of penis and perineum was performed on 30 adult male cadavers (60 sides). Observation and measurement were focused on the penile length of different parts, the morphological relationship of infundibular ligament and suspensory ligament with penile radix, and the feature of crus penis with relation to the deep penile artery. RESULTS: The average length of the penile shaft was 8.13 cm, the penile radix was 7.67 cm and the crus penis was 5.96 - 5.98 cm. The deep penile artery penetrated into the crus penis at its middle 1/3. The infundibular ligament attached to superficial fascia of the penis and extended downward to the scrotal septum to constitute the suspensory structure for both of them. The suspensory ligament attached to the dorsal deep fascia of the penis. Becoming thicker, the rear part of the suspensory ligament connected firmly to the pubic arcuate ligament to constitute a part of suspensory mechanism for the urethra. There was a part of cavernous body, which was free from either ligament or bony attachment, between the penile radix and the crus penis, where the dorsal artery and nerve of penis turned around from the ventral to the dorsal aspect of the penis and the penile dorsal vain penetrated the urogenital septum, draining into intrapelvic venous plexus. CONCLUSIONS: The divisional measurement of the penis length, the recognition of the suspensory ligaments and the anatomic feature of the crus penis with relation to the deep penile artery are all of significant importance to improve the operation of phalloplasty. PMID- 16248524 TI - [Anatomy study of superior and inferior gluteal artery perforator flap]. AB - OBJECTIVE: To find anatomic basis for clinically modifying technique of harvesting superior and inferior gluteal artery perforator flap, in order to avoid muscle lossing in conventional superior and inferior myocutaneous flaps, keep the advantage such as large rich supplied volume soft tissue. METHODS: 5 cases 10 sides adult cadaver were used to study the numbers, position, Course of superior and inferior gluteal artery perforators. The position of perforators was located by ultrasound Doppler in 6 cases and 12 sides in patient's superior and inferior gluteal area. RESULTS: Superior and inferior gluteal artery originated from internal iliac artery. Several main perforators of large caliber were found in the paraischia and central portions of the gluteal muscle, its number was 10 - 15. The length of the vessels varies from 3 to 8 cm and their diameter from 1 - 1.5 mm. These significant perforators pass through the muscle itself and the fascial portion of the muscle to the overlying skin on the gluteal region. The dorsal branches of nervorum lumbalium perforate the deep fascia just above the iliac crest, lateral to the posterior superior iliac spine. If a nerve branch with a substantial diameter crosses the incision line, the nerve can be harvested within the flap. This nerve can be anastomosed to the anterior ramus of the lateral branch of the 4th intercostals nerve. In adult female, 3 - 5 perforators were located by ultrasound Doppler. They distributed in the triangle area among posterior superior iliac crest, the great trochanter and the coccyx. CONCLUSIONS: The area and diameter of perforators of superior gluteal artery were relatively confirmed. It's possible to harvest the perforator flap without any muscle. It has the advantage of conventional myocutaneous flap with out of its disadvantages. It's easy to detect those perforator by ultrasound Doppler clinically. The nerve can be harvested and anastomosed simultaneously. Because the inferior gluteal area is a weight loading area, we suggested to use superior gluteal artery perforator flap. This flap can be transferred pedicled to treat sacral pressure sores or to be transferred freely for the breast reconstruction. PMID- 16248525 TI - [Construction and clinical application of tissue engineered epidermal membrane]. AB - OBJECTIVE: To Construct tissue engineered epidermal membrane for promoting healing of clinical skin graft donor site wound. METHODS: Epidermal cells /Chitosan-Gelatin (CG) membrane was constructed with cultured human epidermal cells(EC) and CG membrane, at the donor site of split skin graft, the wound was divided into three groups: the control group was covered with CG membrane without KC, KC/CG membrane was grafted on the treatment part of the wound area, and blank group was covered with traditional vaseline gauze. The engineered epidermal membrane and its effect on wound were evaluated with gross observation, HE, immunohistochemistry, collagen type I/III ratio by picrosirius polarization method and RT - PCR test at various time points before and after operation. RESULTS: The result showed that human tissue engineered epidermis could be constructed with cultured human EC and CG membrane, and this artificial epidermal membrane could be used for promoting the healing of skin graft donor site wound successfully (16 cases with 3 months' oberservation). The average healing time is (16.2 +/- 3.8) days for control group, (8.1 +/- 1.3) days for experimental group and (23.0 +/- 5.7) days for blank group. The artificial epidermis was well survived with normal structure. In addition, less hypertrophic scar development was observed in treated wound at 90 days (3 in 16 cases, 20.0%) than in the blank sites (11 in 16 cases, 74.4%). The difference is statistically significant (chi2 = 8.127, P < 0.01). CONCLUSIONS: The constructed EC-CG membrane can be constructed in vitro, survived in vivo and has good clinical application in promoting healing of skin graft donor site wound and inhibiting hypertrophic scar formation. PMID- 16248526 TI - [Observe the dissection of the tunnels of augmentation rhinoplasty under endoscope]. AB - OBJECTIVE: To observe and analyse the dissection of the tunnels in traditional blind operation of augmentation rhinoplasty. METHODS: 11 Cases of augmentation rhinoplasty were collected and be observed by an endoscope as soon as the tunnels were formed during the operations. RESULTS: (1) Some of the tunnels did not go through one layer. (2) The bilateral cartilage separated in the mid-line. (3) There were two blood vessels in the surface of alar cartilage. There were perforating blood vessels in the edge of pyriform aperture. (4) In some cases whose incision were in unilateral alar margin, the tunnel were asymmetric. CONCLUSION: In some cases of traditional blind operation of augmentation rhinoplasty, tunnels were not suitable, they were asymmetric; and there were desmo and septa in the tunnels. Those might be the causes of complications post op of augmentation rhinoplasty. PMID- 16248527 TI - [Hair removal with intense pulsed light]. AB - OBJECTIVE: To evaluate the hair-removal effect of intense pulsed light (IPL). METHODS: The unwanted hairs were removed with ELITE plus IPL. The treatment took 3 - 5 procedures, with an interval of over 2 months. 341 patients underwent the treatment. RESULTS: All the patients were well tolerant without anesthesia. The unwanted hairs were wholly removed after 3 - 5 procedures. After operation, routine cleaning and make-up were allowed and bandaging was not necessary. There were blister in 3 cases and infection in 1 case. No pigmentation and scarring happened. Following-up of 3 - 6 months showed steady results with less regeneration of very thin and soft hair. CONCLUSIONS: IPL is an ideal hair removal method because of the credible effect, simple operation, rapid treatment and no serious complications. PMID- 16248529 TI - [Study on using a new biodegradable conduit to repairing rat's peripheral nerve defect]. AB - OBJECTIVE: Nerve conduit, a new way to repair peripheral nerve, has good prospect. Now, the main obstacle of the conduit's clinical using is the absence of material with both ideal physical property and good biocompatibility. In this study, the PLA is added to chitosan to form a novel material used to make a new nerve conduit. And the rat modal is used to get the data of the nerve conduit made of new material to repair the defect of peripheral nerve in vivo. METHODS: Chitosan-PLA Composite Biodegradable material was used to produce a nerve conduit with depth of 200 um, diameter of 1.5 mm. This new conduit was used to repair the sciatic nerve's defect of 5 mm long. The other two groups as control groups repairs the same defects using silicon conduit or nerve's autograft respectively. 12 weeks after operation, normal assessments were performed including movement of hindlimb, hindlimb's retraction after being pricked by needle and ulcer in claw's palm. Special assessment was performed including histology, ECG, image analysis and weighing of triceps muscle of calf. The data of three groups was statistically analyzed by group t test. RESULTS: As the normal observation showing: after 12 weeks, the regenerating nerve succeeded to path through the gap and dominated the muscle. Special assessment including ECG, image analysis and weighing of triceps muscle of calf showed that the group using Chitosan-PLA Composite conduit had the nervous regeneration better than silicon conduit group both in the nerve's quality and in axon's quantity. And the nervous regeneration of Chitosan-PLA Composite conduit group was as good as that of nervous autograft's group. CONCLUSIONS: Chitosan-PLA Composite Biodegradable conduit could repair the defect of peripheral nerves successfully. It is an ideal material for nerve conduit. PMID- 16248528 TI - [Changes of acetylcholine receptor distribution at the motor end-plates following muscle transfer]. AB - OBJECTIVE: To investigate the changes of acetylcholine receptor (AChR) distribution at the neuromuscular junction (i.e. motor end-plate) following the free neurovascular muscle transfer. METHODS: AChR in the gracilis muscle of the Wistar rat following free neurovascular transfer were labeled by fluorescent alpha-bungarotoxin and radioiodinated alpha-bungarotoxin. Then confocal microscope and gamma-counting were estimated to ACHR, qualitatively and quantitatively. RESULTS: The junctional AChR numbers decreased to a minimum at the fourth week postoperatively, whereas the extrajunctional receptor numbers increased. From the fifth week postoperatively, the number of junctional AChR's increased. Even at 30 weeks after transfer, the morphology of the neuromuscular junction failed to return to the preoperative style. The number of acetylcholine receptors at the reinnervated neuromuscular junction also remained lower than the control. CONCLUSION: The persistent weakness following free neurovascular muscle transfer may be attributed to these qualitative and quantitative changes at the neuromuscular junction. PMID- 16248530 TI - [Pathological genomics of keloid fibroblastic cells]. AB - OBJECTIVE: Keloids result from the abnormal repair of the tissues after skin injuries where the pathological overgrowth of large and active fibroblastic cells expands beyond the boundaries of the initiating wound. Imbalanced expression of genes with an as yet unknown regulatory mechanism seems to result in the hypertrophic development of fibroblastic cells and over-productions of collagen. To get information as to genes which function in the actively growing keloid fibroblasts, we have applied a gene expression DNA-microarray technique by analyzing broad range of genes at once in a systematic fashion. METHODS: Differential gene expressions of keloid fibroblastic cell lines against a normal skin fibroblastic cell line, all of the cell lines had been propagated in our lab, were analyzed using a cDNA-microarray technique. mRNA was extracted from the control normal skin cells and the two lines of keloid fibroblastic cells, one from ear-lobe keloid tissue and the other from chest keloid tissue, was subjected to a DNA microarray analysis which includes 1 100 human genes (TaKaRa Intelli Gene Human CHIP 1K Set I) . RESULTS: 8 genes were found to be expressed exclusively in ear-lobe keloid fibroblastic cell lines. Cells from chest keloid were detected to express 17 genes, specifically. Coagulation factor II (thrombin) receptor gene, KIAA0367 protein gene, and matrilin-2 gene were found to be the most commonly expressed genes in the keloid cells. Suppressor genes, like melanoma differentiation associated gene-7, Mda-7 (U16261), were expressed in normal skin fibroblasts but were not expressed in keloid fibroblasts may be implicated in the pathogenesis of the keloid lesions. CONCLUSIONS: Genes expressed specifically in keloid cells may be an adequate pathological diagnostic marker for keloids. Further, Identification of genes that cause cells to develop keloid lesions leads us to gene therapy and prevention of keloids. PMID- 16248531 TI - [Experimental study on induction of the normal human melanocytes apoptosis in vitro by cycloheximide and TNF-alpha]. AB - OBJECTIVE: To investigate the effects of cycloheximide and TNF-alpha on melanocyte (MC). METHODS: Melanocyte apoptosis was studied with MTT, transmitting electron microscopy and fluorescence labeling of alive cells. RESULTS: We added TNF-alpha and cycloheximide in melanocytes, and the typical apoptosis appeared 24 hours later, with chromatin condensation, nuclear pyknosis and apoptotic bodies formation. The results of cytometry showed the typical apoptotic peak. CONCLUSION: TNF-alpha and cycloheximide together could inhibit MC proliferation and induce MC apoptosis. PMID- 16248532 TI - Advances in the value of eggs and egg components for human health. AB - The avian egg is an important source of nutrients, containing all of the proteins, lipids, vitamins, minerals, and growth factors required by the developing embryo, as well as a number of defense factors to protect against bacterial and viral infection. Moreover, eggs are now understood to contain substances with biological functions beyond basic nutrition, and extensive research has been undertaken to identify and characterize these biologically active components. This review mainly focused on biological activities of proteins and peptides derived from egg components. Several biological activities have now been associated with egg components, including novel antimicrobial activities, antiadhesive properties, immunomodulatory, anticancer, and antihypertensive activities, antioxidant properties, protease inhibitors, nutrient bioavailability, and functional lipids, highlighting the importance of egg and egg components in human health and in disease prevention and treatment. Continued research to identify new and existing biological functions of hen egg components will help to define new methods to further improve the value of eggs as a source of numerous biologically active compounds with specific benefits for human and animal health and secure their role in the therapy and prevention of chronic and infectious disease. PMID- 16248533 TI - DNA extraction and analysis from processed coffee beans. AB - The authenticity of coffee is an important issue for both producers and consumers. Premium Arabica material is especially prone to being adulterated, and a number of different techniques have been employed to determine the quality of both roasted and instant coffee. Currently, assessment of coffee authenticity relies on chemical methods which can discriminate between coffee species, but not varieties. Several genetic markers are available for assessing coffee origin, but their suitability to testing commercial coffee is limited by the ability to extract DNA from highly processed beans. In this paper, we demonstrate that PCR grade DNA may be obtained from roasted beans and even instant coffee. This would allow analysis of commercial samples, provided that suitable markers for species/variety identification are found. PMID- 16248534 TI - Simultaneous determination of multiple phytohormones in plant extracts by liquid chromatography-electrospray tandem mass spectrometry. AB - A rapid multiresidue method to quantify three different classes of plant hormones has been developed. The reduced concentrations of these metabolites in real samples with complex matrixes require sensitive techniques for their quantification in small amounts of plant tissue. The method described combines high-performance liquid chromatography with electrospray ionization tandem mass spectrometry. Deuterium-labeled standards were added prior to sample extraction to achieve an accurate quantification of abscisic acid, indole-3-acetic acid, and jasmonic acid in a single run. A simple method of extraction and purification involving only centrifugation, a partition against diethyl ether, and filtration was developed and the analytical method validated in four different plant tissues, citrus leaves, papaya roots, barley seedlings, and barley immature embryos. This method represents a clear advantage because it extensively reduces sample preparation and total time for routine analysis of phytohormones in real plant samples. PMID- 16248535 TI - Determination of tannin in green tea infusion by flow-injection analysis based on quenching the fluorescence of 3-aminophthalate. AB - A flow-injection analytical system was developed to determine tannin content in green tea infusions. The flow-injection system is based on measuring the quenching effect of tannin on the fluorescence of 3-aminophthalate. Fluorophore was obtained by auto-oxidation of luminol during solution preparation. System performance was satisfactory for routine analysis (sample throughput >20 h(-1); linear dynamic range for tannic acid, 0.005-0.3 mg/mL; linear dynamic range for green tea tannin, 0.02-1.0 mg/mL; CV < 3%). The flow-injection method is immune from interference by coexisting ascorbate in green tea infusion. Analytical results were verified by the ferrous tartrate method, the Japanese official analytical method. PMID- 16248537 TI - Study on the supramolecular interaction of thiabendazole and beta-cyclodextrin by spectrophotometry and its analytical application. AB - The beta-cyclodextrin-thiabendazole (beta-CD-TBZ) inclusion complex was synthesized and its structure characterized by (1)H NMR and IR. The mechanism of the supramolecular interaction of TBZ and beta-CD has been studied and discussed by spectrophotometry. The results showed that the phenyl ring of TBZ was included in the beta-CD cavity to form a 1:1 host-guest complex with an apparent formation constant of 1.60 x 10(3) mol(-1).L. On the basis of the enhancement of the absorbance of TBZ produced through complex formation, a spectrophotometric method for the determination of TBZ in bulk aqueous solution in the presence of beta-CD was developed. The linear relationship between the absorbance and TBZ concentration was obtained in the range of 8.86 x 10(-7)-1.45 x 10(-5) mol/L. The detection limit was 2.71 x 10(-7)mol/L, and the relative standard deviation was 0.86%. The interference of 48 coexisting substances was slight. The proposed method has been successfully applied to the determination of TBZ in fruits with recoveries of 96-103%. PMID- 16248536 TI - Production of a monoclonal antibody against ochratoxin A and its application to immunochromatographic assay. AB - A monoclonal antibody (Mab) against ochratoxin A (OTA) was produced from the hybridoma cell line C7G25, which was established by the fusion of Sp2/0-Ag14 myeloma cells with spleen cells isolated from a BALB/c mouse immunized with the OTA-bovine serum albumin conjugate. This Mab belongs to the IgG(2a) heavy-chain subclass with a kappa-type light chain. The level of 50% inhibition concentration was 1.20 ng/mL in a competitive direct enzyme-linked immunosorbent assay (cdELISA), and the detection limit was 0.12 ng/mL. This antibody is specific for OTA but also shows cross-reactivity with ochratoxin B (31.7%) in a cdELISA. On the basis of the sandwich format using the produced Mab against OTA, a rapid immunochromatographic assay was developed to efficiently detect OTA. This method was able to detect up to 500 ng/mL of OTA in <10 min. PMID- 16248538 TI - Genetic diversity and variation of saponin contents in Panax notoginseng roots from a single farm. AB - Radix notoginseng, the root of Panax notoginseng (Burk.) F. H. Chen, has been widely used in traditional Chinese medicine. Its main components, saponins, have been reported to have many pharmacological activities. To test the general assumption that herbs of a single species planted and harvested from a single location are uniform in chemical and genetic makeup, chemical analysis and DNA fingerprinting were carried out. High-performance TLC together with HPLC analysis were used to analyze 17 randomly sampled 3-year-old roots from a single farm for the presence of six saponins. Five roots showed distinct chemical profiles with changed ratios of ginsenosides Rd/Rg1, Re/Rg1, or Rb1/Rg1. The same samples, together with some 1- and 2-year-old samples, were also subjected to fluorescent amplified fragment length polymorphism (AFLP) analysis, and their internal transcribed spacer 2 (ITS 2) regions were sequenced. Fluorescent AFLP analysis was found to be much more polymorphic than the ITS 2 sequence and showed clear evidence of genetic diversity within the tested population. In conclusion, genetic diversity and variation of saponin contents between individual P. notoginseng roots have been detected. We suggest that genetic diversity affects the contents of the six saponins. The saponin contents variation and genetic diversity were also found among P. notoginseng root samples collected from China and Singapore markets. Since variable saponin contents may affect therapeutic efficacy, combining the use of genetic profiling with chemical profiling will help ensure greater uniformity in the quality of P. notoginseng roots. The genetic and chemical diversity within a population also provides the opportunity for breeding new cultivars with more desirable chemical constituents. PMID- 16248539 TI - Monitoring of five sulfonamide antibacterial residues in milk by in-tube solid phase microextraction coupled to high-performance liquid chromatography. AB - A simple, rapid, and sensitive method for the quantitative monitoring of five sulfonamide antibacterial residues in milk was developed by coupling in-tube solid-phase microextraction (SPME) to high-performance liquid chromatography with an ultraviolet detector. A poly(methacrylic acid-ethylene glycol dimethacrylate) monolithic capillary column was selected as the extraction medium for this on line technique. To obtain optimum extraction efficiency, several parameters relating to in-tube SPME were investigated. By simple extraction with ethanol, dilution with phosphate buffer solution, and centrifugation, the sample solution then could be directly injected into the device for extraction. The calculated detection limits for sulfadiazine, sulfamethazine, sulfamethoxazole, sulfamonomethoxine sodium, and sulfacetamide sodium were 2.0, 2.8, 1.7, 2.5, and 22 ng/mL, respectively. The method was linear over the range of 20-5000 ng/mL (100-5000 ng/mL for sulfacetamide sodium) with a correlation coefficient R (2) value >0.9980. Excellent method reproducibility was found by intra- and interbatch precisions, yielding the relative standard deviations of <10.0 and <9.94%, respectively. The proposed method was proved to be robust in monitoring sulfadiazine, sulfamethazine, sulfamethoxazole, sulfamonomethoxine sodium, and sulfacetamide sodium residues in milk. PMID- 16248540 TI - Determination of aflatoxin M1 in milk and milk powder using high-flow solid-phase extraction and liquid chromatography-tandem mass spectrometry. AB - Animal feeds occasionally have some degree of contamination by Aspergillus spp. Even pasteurized milk at times contains the toxic liver carcinogen aflatoxin M1 (AFM1). Confirmation of its presence is now done with solid-phase extraction (SPE) and high-performance liquid chromatography (HPLC)-fluorescence, using a small enough sample that SPE time is reasonable. In this study 200 mL of milk was extracted using a C18 disk at a flow rate of approximately 100 mL/min and AFM1 quantified by HPLC-tandem mass spectrometry with negative electrospray ionization. The effectiveness and cleanup efficacy of immunoaffinity columns (IAC) was compared with that of Mycosep multifunctional cleanup columns (MFC). Average recovery and detection limits of whole milk and low-fat milk cleaned up by IAC were significantly superior to those obtained with the MFC (78-87% and 0.59-0.66 ng/L, respectively). The new procedure improves extraction speed, sensitivity, and specificity. PMID- 16248541 TI - Antiinflammatory properties of the muscadine grape (Vitis rotundifolia). AB - The muscadine grape possesses one of the highest antioxidant levels among fruits; yet, the effect of this fruit on mammalian metabolic systems has not received significant attention. To examine the antiinflammatory properties of the muscadine, grape skins were dried, pulverized, and extracted (10% w/v) with 50% ethanol. The extract was then tested in two different assays: the release of superoxide in phorbol myristate acetate-activated neutrophils and the release of cytokines [tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-beta), and interleukin-6 (IL-6)] by lipopolysaccharide-activated peripheral blood mononuclear cells. The release of superoxide and cytokines was inhibited by increasing concentrations of the extract. A 1:100 dilution of the extract inhibited superoxide release by approximately 60% while the release of TNF-alpha and IL-1beta was reduced at a dilution of 1:200 by approximately 15 and 90%, respectively (all P < 0.05). The inhibition pattern on the release of IL-6 was similar to that seen with TNF-alpha. In a related in vivo study, rats were fed a diet containing 5% (wt/wt) dried muscadine grape skins for 14 days and then were injected with carrageenan in the foot pad. After 3 h, paw edema was measured and the rats on the grape skin diet had approximately 50% less paw edema than controls (P < 0.05). These results demonstrate that the muscadine grape skin powder possesses significant in vitro and in vivo antiinflammatory properties. PMID- 16248542 TI - Catechins and procyanidins in berries of vaccinium species and their antioxidant activity. AB - The fractions of monomeric catechins and the fractions of dimeric and trimeric procyanidins were extracted and concentrated from wild berries of Vaccinium species to study their antioxidant activities. The compositions of the fractions were analyzed using high-performance liquid chromatography combined with diode array and electrospray ionization mass spectrometric detection. Rare A-type dimers and trimers were identified as the predominant procyanidins in wild lingonberry, cranberry, bilberry, and bog whortleberry. Lingonberry and cranberry catechin and procyanidin fractions as well as bog whortleberry catechin fraction were good scavengers of radicals in the 2,2-diphenyl-1-picrylhydrazyl (DPPH) test and more efficient than the respective bilberry fractions. Bog whortleberry procyanidin fraction was less active, this being mainly due to the lower content of these compounds. Fractions from lingonberry, cranberry, and bilberry were equally efficient in inhibiting the oxidation of methyl linoleate emulsion, but differences among the berries were found in their abilities to inhibit low density lipoprotein (LDL) oxidation. Catechins, the monomers, exhibited comparable activity to the fractions containing dimers and trimers in inhibiting the oxidation of methyl linoleate emulsion and human LDL. Bog whortleberry catechins were excellent antioxidants toward the oxidation of human LDL. Radical scavenging and antioxidant activities of Vaccinium berry fractions were attributable to the their composition of catechins and procyanidins. In conclusion, Vaccinium catechins as well as dimeric and trimeric procyanidins provide substantial antioxidant protection. PMID- 16248543 TI - Effect of plant phenolics on protein and lipid oxidation in cooked pork meat patties. AB - Rapeseed and pine bark are rich sources of phenolic compounds that have in previous studies been shown to exhibit antioxidant and anti-inflammatory properties. In this study, the antioxidant effect of rapeseed and pine bark phenolics in inhibiting the oxidation of lipids and proteins in meat was tested as a possible functional food application. The cooked pork meat with added plant material was oxidized for 9 days at 5 degrees C under light. The suitable level of plant material addition was first screened by following lipid oxidation only. For further investigations plant materials were added at a level preventing lipid oxidation by >80%. The oxidation was followed by measuring the formation of hexanal by headspace gas chromatography and the formation of protein carbonyls by converting them to 2,4-dinitrophenylhydrazones and measured by spectrophotometer. It was shown that rapeseed and pine bark were excellent antioxidants toward protein oxidation (inhibitions between 42 and 64%). These results indicate that rapeseed and pine bark could be potential sources of antioxidants in meat products. PMID- 16248544 TI - Effects of population, age, and cultivation methods on ginsenoside content of wild American ginseng (Panax quinquefolium). AB - Genotype and environmental effects on ginsenoside content among eight wild populations of American ginseng (Panax quinquefolium) were investigated. Root concentrations of six ginsenosides were determined at the time of collection of plants from the wild (T0) and 2 years (T2) after transplanting roots from each of the eight populations to each of two different forest garden locations. Both location and population had significant effects on root and shoot growth. Overall, ginsenoside Rb1 was most abundant, followed by Rg1 and Re. Concentrations of Rg1 and Re were inversely related among and within populations. The relative ranking of populations differed depending upon the particular ginsenoside and sampling time. The relative importance of genotype and environment was not the same for all ginsenosides. Ginsenoside Re was influenced by population but not location, whereas Rb1, Rc, and Rb2 were influenced only by location (environment), while Rg1 and Rd were influenced by both. Ginsenoside levels were consistently lower, but growth was consistently higher at the more intensively managed garden location. PMID- 16248545 TI - Flavonoids from Theobroma cacao down-regulate inflammatory mediators. AB - In the present study, we report the effects of a cocoa extract on the secretion and RNA expression of various proinflammatory mediators by macrophages. Monocyte chemoattractant protein 1 and tumor necrosis factor alpha (TNFalpha) were significantly and dose-dependently diminished by cocoa extract, and this effect was higher than that produced by equivalent concentrations of epicatechin but was lower than that produced by isoquercitrin. Interestingly, cocoa extract added prior to cell activation resulted in a significantly greater inhibition of TNFalpha secretion. Both cocoa extract and epicatechin decreased TNFalpha, interleukin (IL) 1alpha, and IL-6 mRNA expression, suggesting that their inhibitory effect on cytokine secretion is produced, in part, at the transcriptional level. Cocoa extract also significantly decreased NO secretion in a dose-dependent manner and with a greater effect than that produced by epicatechin. In conclusion, our study shows that cocoa flavonoids not only inhibit NO release from macrophages but also down-regulate inflammatory cytokines and chemokines. PMID- 16248546 TI - E-cinnamic acid derivatives and phenolics from Chilean strawberry fruits, Fragaria chiloensis ssp. chiloensis. AB - Three E-cinnamic acid glycosides, tryptophan, and cyanidin-3-O-beta-D glucopyranoside were isolated from ripe fruits of the Chilean strawberry Fragaria chiloensis ssp. chiloensis. 1-O-E-Cinnamoyl-beta-D-xylopyranoside, 1-O-E cinnamoyl-beta-D-rhamnopyranoside, and 1-O-E-cinnamoyl-alpha-xylofuranosyl-(1- >6)-beta-D-glucopyranose are reported for the first time. The cinnamic acid glycosides and aromatic compound patterns in F. chiloensis fruits were determined by high-performance liquid chromatography (HPLC). HPLC analyses of extracts showed that cyanidin-3-O-beta-D-glucopyranoside and free ellagic acid are present in achenes while the E-cinnamoyl derivatives and tryptophan were identified only in the thalamus. The free radical scavenging effect of the fruit extract can be associated with the anthocyanin content. PMID- 16248547 TI - Metabolism of glycitein (7,4'-dihydroxy-6-methoxy-isoflavone) by human gut microflora. AB - Gut microbial disappearance and metabolism of the soy isoflavone glycitein, 7,4' dihydroxy-6-methoxyisoflavone, were investigated by incubating glycitein anaerobically with feces from 12 human subjects. The subjects' ages ranged from 24 to 53 years with a body mass index (BMI) of 20.9-25.8 kg/m(2) (mean BMI = 24.0 +/- 1.1 kg/m(2)). Glycitein disappearance followed an apparent first-order rate loss. Fecal glycitein disappearance rates for the subjects segregated into three different groups described as high (k = 0.67 +/- 0.14/h), moderate (k = 0.34 +/- 0.04/h), and low (k = 0.15 +/- 0.07/h) glycitein degraders (p < 0.0001). There was no dose effect on the disappearance rates for each subject from 10 to 250 microM glycitein (average k = 0.32 +/- 0.03/h, p > 0.05). Four putative glycitein metabolites, characterized by liquid chromatography-mass spectrometry (electrospray ionization using positive ionization mode), were dihydroglycitein, dihydro-6,7,4'-trihydroxyisoflavone, and 5'-O-methyl-O-desmethylangolensin. Two subjects produced a metabolite tentatively identified as 6-O-methyl-equol, and one subject produced daidzein as an additional metabolite of glycitein. These results show that glycitein is metabolized by human gut microorganisms and may follow metabolic pathways similar to other soy isoflavones. PMID- 16248548 TI - Protection by petaslignolide A, a major neuroprotective compound in the butanol extract of Petasites japonicus leaves, against oxidative damage in the brains of mice challenged with kainic acid. AB - The neuroprotective effect of petaslignolide A (PA), a furfuran lignan isolated from butanol fraction of Petasites japonicus (Sieb. et Zucc.) Maxim. (Compositae) leaves, on the oxidative damage in the brain of mice challenged with kainic acid was examined using behavioral signs and biochemical parameters of oxidative stress. PA (40 mg/kg) was administered to ICR male mice through a gavage for 4 days consecutively, and on the final day, kainic acid (50 mg/kg) was administered intraperitoneally. During the 4-day treatment with PA, the body weight gain was not significantly different from that of vehicle-treated control animals. PA (40 mg/kg) alleviated the behavioral signs of kainic acid neurotoxicity and reduced the mortality (50%) by kainic acid to 12.5%. Moreover, the administration of PA restored the levels of glutathione and thiobarbituric acid-reactive substances as well as GSH-peroxidase activity in the brains of mice administered kainic acid to control levels (P < 0.05). In comparison, PA (40 mg/kg) was approximately comparable to the butanol fraction (200 mg/kg) of P. japonicus extract in reducing kainic acid neurotoxicity. On the basis of these results, PA is suggested to be a major neuroprotective agent primarily responsible for the protective action of the butanol fraction of P. japonicus extract against kainic acid-induced neurotoxicity in the brains of mice. PMID- 16248549 TI - Free radical scavenging properties and phenolic content of Chinese black-grained wheat. AB - Free radical scavenging properties and phenolic content of extracts from a novel Chinese black-grained wheat were evaluated for comparison with selected wheat controls. Extracts of bran and whole meal were compared for their scavenging activities against the 2,2-diphenyl-1-picryhydrazyl (DPPH) free radical. The total phenolic content and phenolic acid levels were determined using colorimetric and high-performance liquid chromatography (HPLC) methods, respectively. There were significant differences in radical scavenging activities and phenolic contents among bran or whole meal samples of Chinese black-grained wheat and selected wheat controls. Chinese black-grained wheat had the strongest scavenging activity and the highest total phenolic content among the wheat samples. The scavenging activity and total phenolic content of wheat bran was generally twice as high as that of whole meal. A positive correlation was found between DPPH radical scavenging activity and total phenolic content of bran (R = 0.86) and whole meal (R = 0.96). In addition, HPLC analysis detected the presence of gallic, p-hydroxybenzoic, caffeic, syringic, p-coumaric, vanillic, gentisic, o coumaric acid, and ferulic acids in wheat bran. Ferulic acid content was highest among the phenolic acids. Chinese black-grained wheat may be considered as a potential source of natural antioxidants given its high free radical scavenging ability and phenolic content. Additional research is needed to further investigate other phenolic compounds and evaluate their contribution to the antioxidant activity in order to understand the nutraceutical value of the novel black-grained wheat genotype. PMID- 16248550 TI - Effects of naturally occurring compounds on fibril formation and oxidative stress of beta-amyloid. AB - Beta-amyloid (betaA)-induced oxidative toxicity on neuronal cells is a principal route in Alzheimer's disease (AD), and its toxicity occurs after fibril formation. Inhibitory or promoting effects of naturally occurring compounds on betaA fibril formation were evaluated. Among 214 tested compounds, curcuminoids, flavone type flavonoids, and naphthoquinones were shown to be potent inhibitors of betaA fibrilization. The addition of the curcuminoids, curcumin, demethoxycurcumin, and bisdemethoxycurcumin strongly inhibited betaA fibril formation. Flavonoids such as quercetin, rhamnetin, and fisetin strongly inhibited betaA fibril formation. Limonoids, cinnamic acids, and catechins enhanced fibril formation in vitro. Anthothecol possessed the most enhancing activity on fibril formation of the compounds tested. On the other hand, it was found that curcuminoids showed cytotoxicity with the 3-[4,5-dimethylthiazol-2-yl] 2,5-diphenyl tetrazolium bromide assay and did not protect HT22 murine neuroblastoma cells from betaA(25-35) insult. Two flavone type flavonoids, morin and quercetin, exhibited no cytotoxicity and strongly protected HT22 murine neuroblastoma cells from betaA(25-35) oxidative attack. Conclusively, morin or quercetin could be a key molecule for the development of therapeutics for AD. PMID- 16248551 TI - Soy processing affects metabolism and disposition of dietary isoflavones in ovariectomized BALB/c mice. AB - Soy foods and nutritional supplements are widely consumed for potential health benefits. It was previously shown that isoflavone-supplemented diets, which contained equal genistein equivalents, differentially stimulated mammary tumor growth in athymic mice based on the degree of processing. This paper reports plasma pharmacokinetic analysis and metabolite identification using the parental mouse strain fed the same diets, which contained genistin, mixed isoflavones, Novasoy, soy molasses, or soy flour plus mixed isoflavones. Whereas the degree of soy processing did affect several parameters reflecting isoflavone bioavailability and gut microflora metabolism of daidzein to equol, stimulation of tumor growth correlated significantly with only the plasma concentration of aglycon genistein produced by the diets. This conclusion is consistent with the known estrogen agonist activity of genistein aglycon on mammary tumor growth. Conversely, plasma equol concentration was inversely correlated with the degree of soy processing. Although antagonism of genistein-stimulated tumor growth by equol could explain this result, the very low concentration of aglycon equol in plasma (12-fold lower relative to genistein) is inconsistent with any effect. These findings underscore the importance of food processing, which can remove non nutritive components from soy, on the pharmacokinetics and pharmacodynamics of isoflavones. Such changes in diet composition affect circulating, and presumably target tissue, concentrations of genistein aglycon, which initiates estrogen receptor-mediated processes required for the stimulation of tumor growth in a mouse model for postmenopausal breast cancer. PMID- 16248552 TI - Application of 3' untranslated region (UTR) sequence-based amplified polymorphism analysis in the rapid authentication of Radix astragali. AB - Radix astragali (root of Astragalus membranaceus) is an important traditional Chinese medicine. It has been used as a tonic herb for thousands of years in China. The water extract of the roots has a wide range of immunopotentiating effects and has been proven to be efficacious as an adjunct cancer therapy. Authentication of the herbal plant is routinely required for general practice in the field of herbal medicine. To facilitate rapid identification of numerous varieties of Radix astragali that are circulating on the herb markets, a rapid molecular genetic method, named 3' untranslated region (3' UTR) sequence-based amplified polymorphism (UAP), has been developed. A cDNA library was first built from transcripts of an authentic A. membranaceus species. Several cDNA clones specific to A. membranaceus were identified through subtractive hybridization of the A. membranaceus cDNA library with Arabidopsis total cellular RNA. On the basis of these cDNA sequences of the 3' untranslated region (3' UTR) of selected cDNA clones, a Polymerase Chain Reaction (PCR) was performed on genomic DNAs of the dry roots of several putative A. membranaceus. PCR fragment length polymorphism was found between A. membranaceus and its relatives. By using this method, it was possible to differentiate the authentic A. membranaceus root from those putative ones obtained from herbal medicine markets. To the authors' knowledge, this is the first paper applying UAP in the authentication of traditional Chinese medicine plants. PMID- 16248553 TI - Transgenic rice plants expressing human CYP1A1 remediate the triazine herbicides atrazine and simazine. AB - The human cytochrome P450 CYP1A1 gene was introduced into rice plants (Oryza sativa cv. Nipponbare). One-month-old CYP1A1 plants grown in soil clearly showed a healthy growth and tolerance to 8.8 microM atrazine and 50 microM simazine, but nontransgenic plants were completely killed by the herbicides. Although transgenic and nontransgenic plants metabolized the two herbicides into the same sets of compounds, CYP1A1 plants metabolized atrazine and simazine more rapidly than did control plants. In small-scale experiments, residual amounts of atrazine and simazine in the culture medium of CYP1A1 plants were 43.4 and 12.3% of those in control medium; those of nontransgenic Nipponbare were 68.3 and 57.2%, respectively. When cultivated in soil with 2.95 microM atrazine and 3.15 microM simazine for 25 days, CYP1A1 plants eliminated 1.3 times more atrazine and 1.4 times more simazine from the soil than did control plants. Thus, CYP1A1 rice plants make it possible to remove atrazine and simazine more rapidly from the culture medium and soil than can nontransgenic Nipponbare. PMID- 16248554 TI - Fumonisin production by Fusarium verticillioides strains isolated from maize in Mexico and development of a polymerase chain reaction to detect potential toxigenic strains in grains. AB - Fumonisins are mycotoxins produced by Fusarium verticillioides (Sacc. Nirenberg) in maize (Zea mays L.), a staple crop in Mexico. In this study, we report the isolation and identification of 67 Fusarium strains isolated from maize kernels collected in Northwest and Central Mexico. The strains were characterized regarding fumonisin B(1) production and the presence of the FUM1 gene. F. verticillioides was the predominant species isolated in both geographic regions, but the isolates from Northwest Mexico produced higher levels of fumonisin. A polymerase chain reaction (PCR)-based method, to detect a region of the FUM1 gene involved in fumonisin biosynthesis, was developed and employed to detect mycotoxigenic fungi in pure culture and in contaminated maize. The presence of the FUM1 gene was associated with fumonisin production in most isolates, except seven that did not synthesize fumonisin but contained the gene in their genome. The PCR method allowed the direct detection of fungal contamination in ground corn and could be employed to screen for the presence of potential mycotoxigenic fusaria. PMID- 16248555 TI - Behavior of myclobutanil, propiconazole, and nuarimol residues during lager beer brewing. AB - Over a 4 month brewing process, the fate of three fungicides, myclobutanil, propiconazole, and nuarimol, was studied in the spent grain, brewer wort, and final beer product. Only the residual level of myclobutanil after the mashing step was higher than its maximum residue limit (MRL) on barley. A substantial fraction was removed with the spent grain in all cases (26-42%). The half-life times obtained for the fungicides during storage of the spent grains ranged from 82 to 187 days. No significant influence of the boiling stage on the decrease of the fungicide residues was demonstrated. During fermentation, the content reduction varied from 20 to 47%. After the lagering and filtration steps, no significant decrease (<10%) was observed in any of the residues. Finally, during storage of the beer (3 months), the amounts of fungicides fell by 25-50% of their respective concentrations in the finished beer. PMID- 16248557 TI - Influence of the freshness grade of raw fish on the formation of volatile and biogenic amines during the manufacture and storage of vinegar-marinated anchovies. AB - Volatile and biogenic amines from three batches of anchovies, marinated in vinegar, were studied. The anchovies had been vacuum-packed and kept in refrigerated storage for 3 months. Trimethylamine and total volatile basic nitrogen levels were very low and constant throughout marinating and storage process: less than 1 and 10 mg/100 g, respectively. Certain amine levels, mainly those of tyramine and serotonin, increased slightly, particularly during storage. However, even the highest recorded levels were much lower than those considered to be hazardous for human consumption. To study the influence of raw material freshness in the amine profile, two laboratory trials using fresh and spoiled anchovies and simulating the industry standard marinating process were carried out. Levels for both volatile and biogenic amines were dependent on raw material quality, proving consistently higher in those deriving from nonfresh fish. Vinegar marinating leads to a decrease in the accumulation of amines in anchovy while their concentration in the vinegar solution increases due to the vinegar effect as solvent extractor. PMID- 16248556 TI - Physical aging of starch, maltodextrin, and maltose. AB - The physical aging of low water content, amorphous starch/water, maltodextrin/water, and maltose/water mixtures in the glassy state was examined using mechanical testing and calorimetry. Stress relaxation measurements showed that upon storage of the glassy materials there was a time-dependent increase in both flexural modulus and mechanical relaxation time. The mechanical relaxation time increased with depth of quench below the calorimetric glass transition temperature and with aging time at the quench temperature. Calorimetry of the aged materials showed an overshoot in heat capacity in the vicinity of the glass transition. The logarithm of the mechanical relaxation time showed a simple linear relationship with the size of the overshoot expressed as an enthalpy change. The calorimetric behavior could be modeled using the Tool-Narayanaswamy Moynihan method. PMID- 16248558 TI - Electron beam ionization induced oxidative enzymatic activities in pepper (Capsicum annuum L.), associated with ultrastructure cellular damages. AB - Mature green pepper fruits (Capsicum annuum L.) were subjected to ionizing radiation, in the range of 1-7 kGy, with accelerated electrons. Ultrastructural changes by electron microscopy, and the activity of several oxidative metabolism related enzymes such as superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), guaicol peroxidase (POX), and lipoxygenase (LOX), were determined in pericarp tissue just after the ionization treatment and during postionization storage at 7 degrees C followed by 3 days at 20 degrees C. Changes in oxidative stress during the ionization treatment was assessed by the accumulation of malondyaldehide (MDA), a lipid peroxidation product. The ionization induced modifications in the cell ultrastructure, a moderate separation of the plasma membrane from the cell wall being observed for all doses. At 5 and 7 kGy, peroxisomes were not detected and the structures of the chloroplast and vacuoles were seriously damaged. Lipid peroxidation and lipoxygenase activity increased with the ionization dose, staying constant and decreasing, respectively, during the storage period. Conversely, catalase, ascorbate peroxidase, and superoxide dismutase had lower values than in nonionized fruits and, in general, their values did not change or diminished slightly from the seventh day of storage. Peroxidase exhibited an increase in activity with the ionization dose, although these was not a linear relationship, with higher values at 3kGy. Ionization of pepper, especially at doses of 5 and 7 kGy, caused a significant oxidative damage in the fruit, since it increased oxidation and decreased the antioxidant enzymatic defense systems causing ultrastructural changes at cell level. PMID- 16248559 TI - Stabilization of caseinate-covered oil droplets during acidification with high methoxyl pectin. AB - Polysaccharides are widely used in the food industry to modify the stability of protein-based drinks. However, an in depth knowledge of the interactions occurring in the system is still lacking. In this study, the interactions between sodium caseinate and high methoxyl pectin under acidification conditions were studied nondestructively and without dilution using transmission diffusing wave spectroscopy. Oil-in-water emulsions were prepared with 10% soybean oil and 0.5% sodium caseinate. Various concentrations of pectin (ranging from 0 to 0.2%) were added, and the emulsions were acidified with glucono-delta-lactone. With acidification, a "sol-gel" transition occurred and emulsions containing pectin were more stable at lower pH than those without pectin. Furthermore, the sol-gel transition of the mixtures was more sudden for control emulsions without pectin. While in control samples the final solidlike emulsion after gelation tended to be more inhomogeneous and more dissimilar to the starting emulsion, emulsions with pectin in solution gelled later under acidification. With a sufficient amount of pectin, the emulsions showed no aggregation and the destabilization pH varied depending on the amount of pectin present in the emulsions. At intermediate pH values (pH > 5.5), the emulsions displayed a decrease in particle size, more pronounced in samples containing pectin. The results collected using light scattering in concentrated systems, 10% (v/v) in our case, suggested that pectin stabilizes the emulsion oil droplets forming a network of oil droplets loosely connected by strands of pectin. PMID- 16248560 TI - Characterization and quantification of proteins in lecithins. AB - Several methods for extraction and quantification of proteins from lecithins were compared. Extraction with hexane-2-propanol-water followed by amino acid analysis is the most suitable method for isolation and quantification of proteins from lecithins. The detection limit of the method is 15 mg protein/kg lecithin, and the quantification limit is 50 mg protein/kg. The relative repeatability limits for samples containing 0-500 and 500-5000 mg protein/kg sample were 12.6 and 7.5%, respectively. The protein recovery ranged between 101 and 123%. The protein content has been determined in different kinds of lecithins. The results were as follows: standard soy lecithins (between 232 and 1338 mg/kg), deoiled soy lecithin (342 mg/kg), phosphatydylcholine-enriched soy lecithins (not detectable and 163 mg/kg), sunflower lecithins (892 and 414 mg/kg), and egg lecithin (50 mg/kg). The sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein patterns of the standard soy and sunflower lecithins are very similar to those of soy flour. The protein profile of the egg lecithin shows several bands with a broad range of molecular masses. The molecular masses of the main proteins of soy lecithins and soy flour have been determined by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) and ranged from 10.5 to 52.2 kDa. Most of the major proteins from soy and sunflower lecithins identified by MALDI-MS and electrospray tandem MS belong to the 11S globulin fraction, which is one of the main fractions of soy and sunflower seeds. In addition, the seed maturation protein P34 from the 7S globulin fraction of soy proteins has also been identified in soy lecithins. This protein has been reported as the most allergenic protein in soybean. PMID- 16248561 TI - Puerins A and B, two new 8-C substituted flavan-3-ols from Pu-er tea. AB - Pu-er tea is a special treated fermented tea produced from crude green tea, which is prepared from the leaves of Camellia sinensis var. assamica. It is a traditional beverage having been used in China, particularly the southern areas, for a long time. Chemical investigation led to the identification of two new 8-C substituted flavan-3-ols, puerins A (1) and B (2), and two known cinchonain-type phenols, epicatechin-[7,8-bc]-4alpha-(4-hydroxyphenyl)-dihydro-2(3H)-pyranone (3) and cinchonain Ib (4), together with other seven known phenolic compounds, 2,2',6,6'-tetrahydroxydiphenyl (5), (-)-epicatechin-8-C-beta-d-glucopyranoside (6), (-)-epicatechin (EC) (7), (-)-epigallocatechin (EGC) (8), (+/-) gallocatechin (GC) (9), gallic acid (10), and myricetin (11), in addition to caffeine (12). Their structures were determined by spectroscopic methods. The compounds 1-5, which might be formed in the postfermentative process of Pu-er tea, were isolated from tea and theaceous plants for the first time. PMID- 16248562 TI - Antioxidant capacity and phenolic content of spinach as affected by genetics and maturation. AB - Spinach leaves harvested at three maturity stages from eight commercial cultivars (CC) and eight advanced breeding lines (ABL) were evaluated for oxygen radical absorbing capacity (ORAC), total phenolics, and flavonoid composition and content. ABL had higher levels of total phenolics, total flavonoids, and ORAC than CC. Midmaturity spinach leaves had higher levels of total phenolics, total flavonoids, and antioxidant capacity than immature and mature leaves. The contents of individual flavonoids varied in response to maturation, with the predominant glucuronated flavones decreasing and patuletin and spinacetin derivatives increasing. Both total phenolics and total flavonoids correlated well with ORAC (r(xy)() = 0.78 and 0.81, respectively) demonstrating that flavonoids were major contributors to antioxidant capacity. Our results indicate that spinach genotypes should be harvested at the midmaturity stage for consumers to benefit from elevated levels of health promoting flavonoids present in the leaves. Additionally, plant breeders can select for increased phenolic content to increase antioxidant capacity of spinach genotypes. PMID- 16248563 TI - Effects of cyclamen mite (Phytonemus pallidus) and leaf beetle (Galerucella tenella) damage on volatile emission from strawberry (Fragaria x ananassa Duch.) plants and orientation of predatory mites (Neoseiulus cucumeris, N. californicus, and Euseius finlandicus). AB - Volatile emission profile of strawberry (Fragaria x ananassa Duch.) plants (cvs. Polka and Honeoye) damaged by cyclamen mite (Phytonemus pallidus Banks) or leaf beetle Galerucella tenella (L.) (cv. Polka) was analyzed to determine the potential of these strawberry plants to emit herbivore-induced volatiles. The total volatile emissions as well as emissions of many green leaf volatiles (e.g., (Z)-3-hexen-1-ol and (Z)-3-hexenyl acetate) and methyl salicylate were greater from cyclamen mite-damaged strawberry plants than from intact plants. Leaf beetle feeding increased emissions of monoterpenes (Z)-ocimene and (E)-beta-ocimene, sesquiterpenes (E)-beta-caryophyllene, (E,E)-alpha-farnesene, and germacrene-D, and a homoterpene (3E)-4,8-dimethyl-1,3,7-nonatriene (DMNT) significantly. Nevertheless, the naive generalist predatory mites, Neoseiulus cucumeris, Neoseiulus californicus, and Euseius finlandicus did not prefer P. pallidus- or G. tenella-damaged plants over intact plants in a Y-tube olfactometer, suggesting that these predatory mite species are not attracted by the herbivore-induced volatiles being released from young strawberry plants. PMID- 16248564 TI - Influence of carrot psyllid (Trioza apicalis) feeding or exogenous limonene or methyl jasmonate treatment on composition of carrot (Daucus carota) leaf essential oil and headspace volatiles. AB - The effect of carrot psyllid (Trioza apicalis Forster) feeding and limonene and methyl jasmonate (MeJA) treatments on the essential oil composition and headspace volatiles of carrot (Daucus carota ssp. sativus), cvs. Parano and Splendid, leaves was studied. Carrot psyllid feeding significantly increased the concentrations of sabinene, beta-pinene, and limonene, whereas limonene treatment increased the concentration of (Z)-beta-ocimene in the leaves of both cultivars. The limonene treatment significantly increased the concentration of total phenolics in the leaves of both cultivars, and MeJA treatment increased phenolic concentration in the leaves of Parano. Exogenous limonene spray did not decrease the number of carrot psyllid eggs laid either 2 or 24 h after treatment. The results suggest that carrot psyllid feeding induces changes in the endogenous monoterpene pool in the carrot leaves. Limonene and MeJA treatments affect some induced defenses of the carrot, but the exogenous limonene spray is not an effective oviposition deterrent against carrot psyllid. PMID- 16248565 TI - 5-(2,6-difluorobenzyl)oxymethyl-5-methyl-3-(3-methylthiophen-2-yl)- 1,2 isoxazoline as a useful rice herbicide. AB - 5-(2,6-difluorobenzyl)oxymethyl-5-methyl-3-(3-methylthiophen-2-yl)-1,2 isoxazoline derivative was synthesized, and its herbicidal activity was assessed under glasshouse and flooded paddy conditions. 5-(2,6-Difluorobenzyl)oxymethyl-5 methyl-3-(3-methylthiophen-2-yl)-1,2-isoxazoline demonstrated good rice selectivity and potent herbicidal activity against annual weeds at 125 g of a.i. ha(-1) under greenhouse conditions. Soil application of this compound showed complete control of barnyard-grass to the fourth leaf stage at 250 g of a.i. ha( 1). Field trials indicated that this compound controlled annual weeds rapidly with a good tolerance on transplanted rice seedlings by post-emergence and soil application. This compound showed a low mammalian and environmental toxicity in various toxicological tests. PMID- 16248566 TI - Fast and versatile multiresidue method for the analysis of botanical insecticides on fruits and vegetables by HPLC/DAD/MS. AB - A simple multiresidue method for screening analysis of 12 botanical insecticides used by organic farmers has been developed. The method involves a rapid and small scale extraction procedure with acetonitrile. For all fruit and vegetable samples, there was no need for clean up. Rotenone, azadirachtin, ryanodines, and pyrethrins can be separated by high-performance liquid chromatography, quantified, and confirmed with a diode array detector (DAD) and atmospheric pressure chemical ionization mass spectrometry (APCI-MS) in the select ion monitoring mode (SIM). The majority of pesticide recoveries for various fruits and vegetables were >70% in the concentration range from 0.01 to 5 mg/kg. The limit of quantitation for most of the pesticides was 0.01 mg/kg, with the majority of relative standard deviations (RSD) mostly below 10%. PMID- 16248567 TI - Ionic thiocyanate (SCN-) production from 4-hydroxybenzyl glucosinolate contained in Sinapis alba seed meal. AB - Meal produced from Sinapis alba seed by crushing to remove oil contains a glucosinolate that when hydrolyzed produces phytotoxic allelochemicals; however, the responsible compounds and pathways for their production have not been elucidated. S. alba seed meal and partially purified extracts containing 4 hydroxybenzyl glucosinolate were included in experiments to identify and monitor enzymatically released products using GC-MS and HPLC-MS. The initial product, 4 hydroxybenzyl isothiocyanate, was unstable in aqueous media, showing a half-life of 321 min at pH 3.0, decreasing to 6 min at pH 6.5. More alkaline pH values decrease the stability of 4-hydroxybenzyl isothiocyanate by promoting the formation of a proposed quinone that hydrolyzes to SCN-. Measurement of SCN- showed stoichiometric release from S. alba meal at 48 h when buffered at pH values as low as 4.0, demonstrating that SCN- production in soil is not only probable but likely responsible for observed phytoxicity of the meal. PMID- 16248568 TI - Efficiency of rice bran for the removal of selected organics from water: kinetic and thermodynamic investigations. AB - The sorption efficiency of indigenous rice (Oryza sativa) bran for the removal of organics, that is, benzene, toluene, ethylbenzene, and cumene (BTEC), from aqueous solutions has been studied. The sorption of BTEC by rice bran is observed over a wide pH range of 1-10, indicating its high applicability to remove these organics from various industrial effluents. Rice bran effectively adsorbs BTEC of 10 microg mL(-1) sorbate concentration from water at temperatures of 283-323 +/- 2 K. The effect of pH, agitation time between solid and liquid phases, sorbent dose, its particle size, and temperature on the sorption of BTEC onto rice bran has been studied. The pore area and average pore diameter of rice bran by BET method are found to be 19 +/- 0.7 m(2) g(-1) and 52.8 +/- 1.3 nm. The rice bran exhibits appreciable sorption of the order of 85 +/- 3.5, 91 +/- 1.8, 94 +/- 1.4, and 96 +/- 1.2% for 10 microg mL(-1) concentration of benzene, toluene, ethylbenzene, and cumene, respectively, in 60 min of agitation time using 0.1 g of rice bran at pH 6 and 303 K. The sorption data follow Freundlich, Langmuir, and Dubinin-Radushkevich (D-R) models. Sorption capacities have been computed for BTEC by Freundlich (32 +/- 3, 61 +/- 14, 123 +/- 28, and 142 +/- 37 m mol g(-1)), Langmuir (6.6 +/- 0.1, 7.5 +/- 0.13, 9.5 +/- 0.22, and 9.4 +/- 0.18 m mol g(-1)), and D-R isotherms (11 +/- 0.5, 16 +/- 1.3, 30 +/- 2.2, and 33 +/- 2.5 m mol g( 1)), respectively. The Lagergren equation is employed for the kinetics of the sorption of BTEC onto rice bran and first-order rate constants (0.03 +/- 0.002, 0.04 +/- 0.003, 0.04 +/- 0.003, and 0.05 +/- 0.004 min(-1)) have been computed for BTEC at their concentration of 100 mug mL(-1) at 303 K. Studies on the variation of sorption with temperatures (283-323 K) at 100 mug mL(-1) sorbate concentration gave thermodynamic constants DeltaH (kJ mol(-1)), DeltaG (kJ mol( 1)), and DeltaS (J mol(-1) K(-1)). The results indicate that the sorption of organics onto rice bran is exothermic and spontaneous in nature under the optimized experimental conditions selected. This sorbent has been used successfully to accumulate and then to determine benzene, toluene, and ethylbenzene in wastewater sample. PMID- 16248569 TI - Multidimensional gas chromatography-olfactometry for the identification and prioritization of malodors from confined animal feeding operations. AB - Odor profiling efforts were directed at applying to high-density livestock operations some of the lessons learned in resolving past, highly diverse, odor focused investigations in the consumer product industry. Solid-phase microextraction (SPME) was used for field air sampling of odorous air near and downwind of a beef cattle feedyard and a swine finisher barn in Texas. Multidimensional gas chromatography-olfactometry (MDGC-O) was utilized in an attempt to define and prioritize the basic building blocks of odor character associated with these livestock operations. Although scores of potential odorant volatiles have been previously identified in high-density livestock operations, the odor profile results developed herein suggest that only a very few of these may constitute the preponderance of the odor complaints associated with these environments. This appeared to be especially true for the case of increasing distance from both cattle feedyard and swine barn facilities, with p-cresol consistently taking on the dominant odor impact role with ever increasing distance. In contrast, at- or near-site odor profiles were shown to be much more complex, with many of the well-known lower tier odorant compounds rising in relative significance. For the cattle feedyard at- or near-site odor profiles, trimethylamine was shown to represent a significantly greater individual odor impact relative to the more often cited livestock odorants such as hydrogen sulfide, the organic sulfides, and volatile fatty acids. This study demonstrates that SPME combined with a MDGC-O-mass spectrometry system can be used for the sampling, identification, and prioritization of odors associated with livestock. PMID- 16248570 TI - Methodological approach to the study of the formation and physicochemical properties of phosphate-metal-humic complexes in solution. AB - The aim of this work is to study the suitability of the complementary use of ultrafiltration (UF) and the interaction with an anion-exchange resin (AR) to characterize of phosphate-metal-humic complexes in solution. The results indicate that a methodological approach consisting of the validation and calibration of the AR method by the UF method and the further use of the AR method is suitable for characterizing phosphate-metal complexes. Such an approach has proven to be useful for calculating the phosphate maximum binding capacity of iron-humic complexes and stability constants. It might also be used to obtain valuable purified phosphate-metal-humic complexes for further structural characterization. PMID- 16248571 TI - Within-season volatile and quality differences in stored fresh-cut cantaloupe cultivars. AB - Cantaloupe cultivar variability from various U.S. regions and growing seasons within a given year was evaluated. As expected, there was often considerable quality and volatile variation among cultivars. Eight of 11 cultivars met the standard U.S. No. 1 requirement for degrees Brix (> or =9), and in most cultivars, degrees Brix declined during fresh-cut storage at 4 degrees C. Hunter L (loss of lightness color value) and a (decline of typical orange hue) colors also generally declined during storage in most cultivars. Volatile ester compounds generally decreased during fresh-cut storage or exhibited a transient increase before declining after 5-7 days of storage. The relative percentage of acetate esters declined during storage in all cultivars, and declines were accompanied by simultaneous non-acetate ester increases. Slight imbalances in compound concentrations may alter the overall perception of desirable, typical "cantaloupe" aroma/flavor during fresh-cut storage. Upsetting the unique aroma balance through storage may negatively affect flavor and the consumer's perception of desirable attributes, even though total volatile levels might not decrease substantially until after 5-7 days in storage. Subtle volatile and quality decreases are likely to be exacerbated with immature-harvested cantaloupe and are likely in out-of-season exports that have likewise been harvested at less mature stages. Altogether, this study indicates the difficulty in procuring cantaloupes of consistent quality from local producers, in a given year, for domestically grown fruit. PMID- 16248572 TI - Flavor characteristics of lapsang souchong and smoked lapsang souchong, a special Chinese black tea with pine smoking process. AB - The major volatile constituents of lapsang souchong, smoked lapsang souchong, and smoked souchong, a group of special black teas in China, were analyzed with GC and GC-MS analyses. Forty-nine constituents were identified. Longifolene and alpha-terpineol were the most abundant compounds in the aroma. Due to its special production process, the compounds longifolene, longicyclene, guaiacol, 4 methylguaiacol, 4-ethylguaiacol, etc., were identified only in this kind of black tea. The aroma constituents of tea origin decreased during the smoking process, whereas the pine terpenoids and the thermal pyrolysis products of the pine wood increased markedly. The pine material for smoking was also analyzed for the volatile constituents showing apparent selectivity of the tea leaves to absorb. There were apparent differences among the tea samples in the contents of aroma constituents. The characteristics of the aroma of the teas are discussed with their processes and the features of sensory qualities. PMID- 16248573 TI - Composition and Antimicrobial Activity of Anemopsis californica leaf oil. AB - Isolation and characterization of leaf volatiles in Anemopsis californica (Nutt.) Hook. and Arn. (A. californica) was performed using steam distillation, solid phase microextraction, and supercritical fluid extraction. Thirty-eight compounds were detected and identified by gas chromatography; elemicin was the major component of the leaf volatiles. While the composition of the leaf volatiles varied with method of extraction, alpha-pinene, sabinene, beta-phellandrene, 1,8 cineole, piperitone, methyl eugenol, (E)-caryophyllene, and elemicin were usually present in readily detectable amounts. Greenhouse-reared clones of a wild population of A. californica had an identical leaf volatile composition with the parent plants. Steam-distilled oil had antimicrobial properties against 3 (Staphylococcus aureus, Streptococcus pneumoniae, and Geotrichim candidum) of 11 microbial species tested. Some of this bioactivity could be accounted for by the alpha-pinene in the oil. PMID- 16248574 TI - Characterization of (E,E,Z)-2,4,6-nonatrienal as a character impact aroma compound of oat flakes. AB - To identify the compounds evoking the characteristic cereal-like, sweet aroma of oat flakes, an aroma extract dilution analysis (AEDA) was applied to a distillate prepared by solvent extraction/vacuum distillation from commercial oat flakes. Among the nine aroma-active compounds detected by gas chromatography-olfactometry and AEDA in the flavor dilution (FD) factor range of 4-1024, eight odorants, for example, (E)-beta-damascenone, (Z)-3-hexenal, and butanoic acid, showed only low FD factors. However, one odorant eliciting the typical cereal, sweet aroma of the flakes was detected with the highest FD factor of 1024. By mass spectrometry and nuclear magnetic resonance measurements followed by a synthesis, (E,E,Z)-2,4,6 nonatrienal, exhibiting an intense oat flake-like odor at the extremely low odor threshold of 0.0002 ng/L in air, was identified as the key odorant of the flakes. By means of a newly developed stable isotope dilution analysis using synthesized, carbon-13-labeled nonatrienal as the internal standard, a concentration of 13 mug of (E,E,Z)-2,4,6-nonatrienal per kilogram of the flakes was measured. Model studies suggested linolenic acid as the precursor of nonatrienal in oats. PMID- 16248575 TI - Effect of some phenolic compounds and beverages on pepsin activity during simulated gastric digestion. AB - The effect of some polyphenols (resveratrol, catechin, epigallocatechin-3 gallate, and quercetin) and beverages (red wine and green tea) on the enzymatic activity of pepsin during the digestion of three different substrates (pork meat, insoluble azocasein, and denatured hemoglobin) has been investigated. The tested polyphenols and beverages increase the initial velocity of the reaction, and the activating effect is concentration dependent. The order of effectiveness of polyphenols in increasing the initial velocity of the reaction is resveratrol > or = quercetin > epigallocatechin-3-gallate > catechin. The kinetic data obtained with soluble denatured hemoglobin show that the K(m) for the substrate is not changed, whereas the V(max) of the reaction is increased. Pepsin activity follows a simple Michaelis-Menten kinetic suggesting that k(3) is increased by polyphenols. To the authors' knowledge, the present study is the first demonstration that some polyphenols and related beverages are able to enhance the enzymatic activity of pepsin. PMID- 16248576 TI - Enzymatic hydrogenation of trans-2-nonenal in barley. AB - Conversion of undesirable, taste-active compounds is crucial for using barley as a suitable raw material for beer production. Here, ALH1, a barley alkenal hydrogenase enzyme that reduced the alpha,beta-unsaturated double bond of aldehydes and ketones, was found to convert trans-2-nonenal (T2N), a major contributor to the cardboard-like flavor of aged beer. Although the physiological function of ALH1 in barley development remains elusive, it exhibited high specificity with NADPH as a cofactor in the conversion of several oxylipins including T2N, trans-2-hexenal, traumatin, and 1-octen-3-one. ALH1 action represents a previously unknown mechanism for T2N conversion in barley. Additional experimental results resolved the genomic sequence for barley ALH1, as well as the identification of a paralog gene encoding ALH2. Interestingly, T2N was not converted by purified, recombinant ALH2. The possibility to enhance ALH1 activity in planta is discussed--not only with respect to the physiological consequences thereof--but also in relation to improved beer quality. PMID- 16248577 TI - Ultrasonic spectroscopy and differential scanning calorimetry of liposomal encapsulated nisin. AB - The thermal stability of phosphatidylcholine (PC) liposomes (colloidal dispersions of bilayer-forming polar lipids in aqueous solvents) in the presence and absence of the antimicrobial polypeptide nisin was evaluated using differential scanning calorimetry (DSC) and low-intensity ultrasonic spectroscopy (US). PC liposome mixtures with varying acyl chain lengths (C16:0 and C18:0) were formed in buffer with or without entrapped nisin. Gel-to-liquid crystalline phase transition temperatures (T(M)) of liposomes determined from DSC thermograms were in excellent agreement with those determined by ultrasonic velocity and attenuation coefficient measurements recorded at 5 MHz. The dipalmitoylphosphatidylcholine (DPPC) T(M) measured by DSC was approximately 41.3 and approximately 40.7 degrees C when measured by ultrasonic spectroscopy. The T(M) of distearoylphosphatidylcholine (DSPC) and DPPC/DSPC 1:1 liposomes was 54.3 and 54.9 degrees C and approximately 44.8 and approximately 47.3 degrees C when measured by DSC and US, respectively. The thermotropic stability generally increased upon addition of nisin. Analysis of the stepwise decrease in ultrasonic velocity with temperature indicated an increased compressibility corresponding to a loss of structure upon heating. PMID- 16248578 TI - Wine volatile and amino acid composition after malolactic fermentation: effect of Oenococcus oeni and Lactobacillus plantarum starter cultures. AB - Red wine amino acids and volatile compounds were analyzed before and after malolactic fermentation carried out by four different starter cultures of the species Oenococcus oeni and Lactobacillus plantarum. The purpose of this study was to determine whether differences can be attributed to the lactic acid bacteria strain used in this important step of the wine-making process. The malolactic cultures selected for this study were indigenous wine lactic acid bacteria strains. The data were evaluated using different multivariate analysis techniques. Results showed different malolactic behaviors for O. oeni and L. plantarum and significant metabolic differences between both species. A degree of diversity was found within each lactic acid bacteria group, since wines presented specific characteristics depending on the lactic acid bacteria strain used. In all cases, malolactic fermentation seemed to modify the amino acid and volatile composition of the wine. PMID- 16248579 TI - Evaluation of the solubility and emulsifying property of soybean proglycinin and rapeseed procruciferin in relation to structure modified by protein engineering. AB - The presence or absence of a highly negatively charged extension region in beta conglycinin (J. Agric. Food Chem. 1999, 47, 5278) and the length of a highly negatively charged variable region IV in glycinin (J. Agric. Food Chem. 2004, 52, 8197) are important determinants of solubility and emulsifying property. To examine the effects of the variable region IV from proglycinin A1aB1b and A3B4 and of the extension region from beta-conglycinin alpha' (alpha'ext) on solubility and emulsifying properties in detail, several mutants of proglycinin, procruciferin, and beta-conglycinin were designed and prepared in Escherichia coli. Nine out of 10 mutants were expressed at high levels in E. coli and shown to be homotrimer similar to the wild types as assessed by gel filtration. The position of the introduced negatively charged region as well as the amino acid composition were demonstrated to affect solubility at mu = 0.08. All of the proglycinin, procruciferin, and beta-conglycinin mutants with the alpha'ext in the C-terminus, especially the proglycinin mutant, exhibited excellent emulsifying ability and emulsion stability. These indicate that improvement of emulsifying properties by insertion of the alpha'ext in the C-terminus may be generally applicable to seed globulins. PMID- 16248580 TI - Ultrastructure of individual and compound starch granules in isolation preparation from a high-quality, low-amylose rice, ilpumbyeo, and its mutant, G2, a high-dietary fiber, high-amylose rice. AB - The ultrastructures of isolated starch granules from Ilpumbyeo (IP), a low amylose japonica rice, and its mutant, Goami2 (G2), a high-amylose rice, which have extreme contrasts in physicochemical properties, cooking qualities (Kang, H. J.; Hwang, I. K.; Kim, K. S.; Choi, H. C. Comparative structure and physicochemical properties of Ilpumbyeo, a high-quality japonica rice, and its mutant, Suweon 464. J. Agric. Food Chem. 2003, 51, 6598-6603. Kim, K. S.; Kang, H. J.; Hwang, I. K.; Hwang, H. G.; Kim, T. Y.; Choi, H. C. Comparative ultrastructure of Ilpumbyeo, a high-quality japonica rice, and its mutant, Suweon 464: Scanning and transmission electron microscopy studies. J. Agric. Food Chem. 2004, 52, 3876-3883), and susceptibility to amylolytic enzymes (Kim, K. S.; Kang, H. J.; Hwang, I. K.; Hwang, H. G.; Kim, T. Y.; Choi, H. C. Fibrillar microfilaments associated with a high-amylose rice, Goami2, a mutant of Ilpumbyeo, a high-quality japonica rice. J. Agric. Food Chem. 2005, 53, 2600 2608), were compared. In isolated preparation, IP consisted entirely of well separated individual starch granules (ISG), whereas G2 consisted of two populations, the large voluminous bodies and the smaller forms, the ISGs. High voltage electron microscopy revealed that each of the voluminous bodies consisted of tightly packed smaller subunits, the ISGs, indicating that they represent the compound starch granules (CSGs) of G2. This suggests that the structural as well as functional unit of G2 involved in food processing is, unlike IP and other ordinary rices, not ISG but is primarily CSG. ISGs located at the periphery of CSGs were fused to each other with adjacent ones forming a thick band or wall encircling the entire circumference. The periphery of ISGs separated from CSGs of G2 consisted of thin radially oriented filaments arranged side by side along the entire granule surface, whereas no such filaments occurred in ISG of IP. It appears that the thick band and the peripheral filaments surrounding CSGs and ISGs, respectively, function as a structural barrier that limits the entrance of water into the granules and subsequent absorption, causing the low swelling power, incomplete gelatinization, and finally poor quality of cooked rice in G2. PMID- 16248581 TI - Common physical-chemical properties correlate with similar structure of the IgE epitopes of peanut allergens. AB - Although many sequences and linear IgE epitopes of allergenic proteins have been identified and archived in databases, structural and physicochemical discriminators that define their specific properties are lacking. Current bioinformatics tools for predicting the potential allergenicity of a novel protein use methods that were not designed to compare peptides. Novel tools to determine the quantitative sequence and three-dimensional (3D) relationships between IgE epitopes of major allergens from peanut and other foods have been implemented in the Structural Database of Allergenic Proteins (SDAP; http://fermi.utmb.edu/SDAP/). These peptide comparison tools are based on five dimensional physicochemical property (PCP) vectors. Sequences from SDAP proteins similar in their physicochemical properties to known epitopes of Ara h 1 and Ara h 2 were identified by calculating property distance (PD) values. A 3D model of Ara h 1 was generated to visualize the 3D structure and surface exposure of the epitope regions and peptides with a low PD value to them. Many sequences similar to the known epitopes were identified in related nut allergens, and others were within the sequences of Ara h 1 and Ara h 2. Some of the sequences with low PD values correspond to other known epitopes. Regions with low PD values to one another in Ara h 1 had similar predicted structure, on opposite sides of the internal dimer axis. The PD scale detected epitope pairs that are similar in structure and/or reactivity with patient IgE. The high immunogenicity and IgE reactivity of peanut allergen proteins might be due to the proteins' arrays of similar antigenic regions on opposite sides of a single protein structure. PMID- 16248582 TI - Contrasting effects of puerarin and daidzin on glucose homeostasis in mice. AB - Puerarin and daidzin are the major isoflavone glucosides found in kudzu dietary supplements. In this study, we demonstrated that puerarin significantly improves glucose tolerance in C57BL/6J-ob/ob mice, an animal model of type 2 diabetes mellitus, blunting the rise in blood glucose levels after i.p. administration of glucose. In contrast, daidzin, the O-glucoside, had a significant but opposite effect, impairing glucose tolerance as compared to saline-treated controls. When they were administered i.p. with (14)C-glucose to C57BL/6J lean mice, puerarin inhibited glucose uptake into tissues and incorporation into glycogen, while daidzin stimulated glucose uptake, showing an opposite effect to puerarin. Puerarin also antagonized the stimulatory effect of decyl-beta-D-thiomaltoside, an artificial primer of glycogen synthesis, which increases (14)C-glucose uptake and incorporation into glycogen in mouse liver and heart. A liquid chromatography tandem mass spectrometry procedure was used to investigate the metabolism and bioavailability of puerarin and daidzin. The blood puerarin concentration-time curve by i.p. and oral administration indicated that puerarin was four times more bioavailable via i.p. injection than via the oral route of administration. This may account for the increased hypoglycemic effect seen in the i.p. glucose tolerance test vs that seen orally. Our results suggest that puerarin is rapidly absorbed from the intestine without metabolism, while daidzin is hydrolyzed to the aglycone daidzein. The opposing effects of puerarin and daidzin on glucose homeostasis may have implications for the activity of dietary supplements that contain both of these isoflavonoids. PMID- 16248583 TI - Hydrogen peroxide induced oxidative stress damage and antioxidant enzyme response in Caco-2 human colon cells. AB - Studies were conducted to evaluate the cell damage caused by exposing human colon carcinoma cells, Caco-2, to hydrogen peroxide at concentrations varying from 0 to 250 microM for 30 min. Evaluation of cell viability, as measured by trypan blue dye exclusion test, showed that the loss of viability was < 5% at concentrations up to 250 microM hydrogen peroxide. Cell membrane damage and DNA damage as measured by the leakage of lactate dehydrogenase and the comet assay, respectively, were significantly high at concentrations >100 microM hydrogen peroxide compared to those of the control. Antioxidant mechanisms in Caco-2 cells were evaluated by measuring catalase, superoxide dismutase, and glutathione peroxidase activities. Catalase activities remained constant in cells treated with 50-250 microM hydrogen peroxide. Superoxide dismutase activity decreased, whereas glutathione peroxidase activity increased in cells treated with H(2)O(2) concentrations of >50 microM. This study showed that with increasing hydrogen peroxide concentration, cell membrane leakage and DNA damage increased, whereas the three antioxidant enzymes responded differently, as shown by mathematical models. PMID- 16248584 TI - Characterization of anthocyanin-rich waste from purple corncobs (Zea mays L.) and its application to color milk. AB - Pigment production from anthocyanin-rich purple corncobs generates a deeply colored waste precipitate. Our objectives were to characterize this anthocyanin rich waste (ARW) and to find a suitable application in a food matrix. Composition and solubility characteristics of ARW were evaluated. Color (CIELAB) and pigment (monomeric anthocyanin and HPLC profiles) stability of ARW in milk (35 mg/100 mL) were evaluated using an accelerated test at 70 degrees C and phosphate buffer as a control. ARW provided milk an attractive purple hue (324-347 degrees ). Monomeric anthocyanin degradation followed zero-order kinetics in skim and whole milk and second-order kinetics in the control, with half-lives of 173, 223, and 44 min at 70 degrees C, respectively. ARW shows potential as a natural colorant for a pH range unusual for anthocyanin applications. A protective effect of matrix constituents on the stability of anthocyanins was evident. Anthocyanins may interact with different compounds in biological systems when the pH values are close to neutral. PMID- 16248585 TI - Determination of the grape invertase content (using PTA-ELISA) following various fining treatments versus changes in the total protein content of wine. relationships with wine foamability. AB - Proteins have proven to play a major role in the stabilization of foam in Champagne wines despite their low concentration that ranges from 4 to 20 mg/L. The aim of this study was to evaluate the effect of fining on total protein and grape invertase contents of champenois base wines and their foaming properties. Data showed that fining and especially the use of bentonite at doses ranging from 10 to 50 g/hL leads to a significant decrease in the total protein content of wines together with that of the grape invertase content, with such a decrease being very detrimental to the foaming properties of the treated wines in terms of foam height (HM) and foam stability (HS). Only a slight decrease in the total protein content, in the grape invertase concentration, and in the foam quality of wines was observed when using casein (10 and 20 g/hL) or bentonite combined with casein (both at 20 g/hL). Our study thus clearly establishes the good correlation existing between the wine protein concentration and its foaming properties. A remarkable correlation was observed between the decrease in the grape invertase content and the total protein content of wines, following bentonite treatments, suggesting that the grape invertase (which represents at least 10-20% of the wine proteins) follows a similar behavior upon fining to other proteins of Champagne wines, despite the high molecular mass and the highly glycosylated structure of this particular protein. Moreover, the decrease in total protein and grape invertase contents of wine after fining with bentonite was found to be correlated with a decrease in the foaming properties of the corresponding wines (with respectively R(2) = 0.89 and 0.95). PMID- 16248587 TI - In vitro bile acid binding of flours from oat lines varying in percentage and molecular weight distribution of beta-glucan. AB - Two experimental high beta-glucan oat (Avena sativa) lines (7.64 and 8.05%) and two traditional lines (4.77 and 5.26% beta-glucan) were used to evaluate the effect of beta-glucan quantity and molecular weight on bile acid (BA) binding. The oat flour samples were digested by an in vitro system that simulated human digestion. No significant differences among oat type were found in the overall beta-glucan, starch, and pentosan digestibilities. Considering the standard, cholestyramine, as 100% bound, the relative BA binding for the oat flour samples on a dry matter basis was in the range of 7.5-14.8%, which is higher than the values determined for some other grains and plant materials in the literature. Although the high beta-glucan flours bound a high amount of BA, no significant correlations were found between beta-glucan content in the flours and BA binding. Significant correlations were found between BA binding and insoluble dietary fiber content. Partial hydrolysis with lichenase of the beta-glucan molecules did not affect the BA binding. A summary of all data suggested that BA binding is a multicomponent-dependent process. PMID- 16248586 TI - Carbohydrate metabolism as related to high-temperature conditioning and peel disorders occurring during storage of citrus fruit. AB - The aim of this research was to understand the involvement of the carbohydrate metabolism in physiological disorders occurring during the postharvest storage of citrus fruit. These disorders, manifested in the rind, depreciate fruit quality and often originate important losses. There has been increasing interest in the use of nonharmful treatments, such as high-temperature conditioning, to avoid citrus peel damage during fruit storage at low temperature in chilling-sensitive cultivars, but their influence in postharvest disorders occurring at nonchilling temperatures and the mechanisms related to them are poorly understood. The data obtained showed that heat conditioning (3 days/37 degrees C) increases the chilling tolerance of cv. Navelate fruit and favored sucrose, but not hexoses, accumulation and its maintenance after the fruit was transferred to low temperature. This effect was related to heat-induced increase in the activities of the sucrose-synthesizing enzymes sucrose phosphate synthase (SPS) and sucrose synthase (SS). Furthermore, sucrose levels and the activities of both enzymes were higher in cv. Pinalate oranges, a chilling-tolerant spontaneous abscisic acid deficient mutant of Navelate. In contrast, carbohydrates appeared not to be involved in the susceptibility of oranges to rind staining, a physiological disorder different from chilling injury, which mainly occurred at a nonchilling temperature (12 degrees C) and was not reduced by heat conditioning. The effect of low temperature in SS and SPS activities was less than that of high temperature, which might be related to the lower changes occurring in sucrose during fruit storage at 2 degrees C. PMID- 16248588 TI - Study of anticancer activities of muscadine grape phenolics in vitro. AB - Muscadine grapes have unique aroma and flavor characteristics. Although a few studies reported high polyphenols content of muscadine grapes, little research has been conducted to evaluate the phenolic compounds bioactivities in any muscadine grape cultivar. The objective of this study was to evaluate the effect of phenolic compounds in muscadine grapes on cancer cell viability and apoptosis. Four cultivars of muscadine (Carlos, Ison, Noble, and Supreme) were assessed in this study. Phenolic compounds were extracted from muscadine skins and further separated into phenolic acids, tannins, flavonols, and anthocyanins using HLB cartridge and LH20 column. Some individual phenolic acids and flavonoids were identified by HPLC. Anthocyanin fractions were more than 90% pure. The effect of different fractions on the viability and apoptosis of two colon cancer cell lines (HT-29 and Caco-2) was evaluated. A 50% inhibition of cancer cell population growth for the two cell lines was observed at concentrations of 1-7 mg/mL for crude extracts. The phenolic acid fractions showed a 50% inhibition at the level of 0.5-3 mg/mL. The greatest inhibitory activity was found in the anthocyanin fraction, with a 50% inhibition at concentrations of approximately 200 microg/mL in HT-29 and 100-300 microg/mL in Caco-2. Anthocyanin fractions also resulted in 2-4 times increase in DNA fragmentation, indicating the induction of apoptosis. These findings suggest that polyphenols from muscadine grapes may have anticancer properties. PMID- 16248589 TI - Organoarsenical species contents in cooked seafood. AB - The organoarsenical species arsenobetaine (AB), arsenocholine (AC), tetramethylarsonium ion (TMA+), dimethylarsinic acid (DMA), and monomethylarsonic acid (MMA) were determined in 64 cooked seafood products (fish, bivalves, squid, crustaceans) included in a Total Diet Study carried out in the Basque Country (Spain). For cooking, various treatments were employed (grilling, roasting, baking, stewing, boiling, steaming, microwaving). The results obtained show that in cooked seafood AB is the major species, followed by DMA and TMA+. AC and MMA are minor species. The results in cooked seafood were compared with the arsenic species contents obtained for the same product raw. After cooking there was an increase in DMA for sardines and bivalves and an increase or appearance of TMA+ for meagrim, anchovy, Atlantic horse mackerel, and sardine. The data provided add to the very scant information available about organoarsenical species contents in cooked seafood. PMID- 16248590 TI - Effects of cooking on concentrations of polychlorinated dibenzo-p-dioxins and related compounds in fish and meat. AB - We investigated the cooking-induced changes in concentrations of polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs), and dioxin-like polychlorinated biphenyls (PCBs) (dioxins) using mackerel and beef. The concentrations of dioxins (29 congeners) were determined by isomer specific analyses and were compared between uncooked and cooked samples. The cooking procedures examined in this study included grilling as a fillet, boiling as a fillet, and boiling as tsumire (small, hand-rolled balls) for mackerel and boiling as a slice, broiling as a slice, and broiling as a hamburger for beef. Three trials were carried out for each cooking method. Generally, concentrations of dioxins were reduced in every cooking trial. When nondetected congener concentrations were assumed to be half the limit of detection for mackerel, the maximum percentage reductions of total concentrations given as 2,3,7,8-tetraCDD equivalents (TEQ) were 31% in grilling as a slice, 14% in boiling as a slice, and 21% in boiling as tsumire under the conditions of this study. In contrast, for beef, the reductions were 42% in boiling as a slice, 42% in broiling as a slice, and 44% in broiling as a hamburger. These results suggest that ordinary cooking processes with heating undoubtedly reduce the dioxin content in animal products, and the reductions estimated should be considered when dioxin intake is evaluated using contamination data for individual food items. PMID- 16248591 TI - Bioavailability of inorganic arsenic in cooked rice: practical aspects for human health risk assessments. AB - Arsenic is present in rice grain mainly as inorganic arsenic. Little is known about the effect of cooking on inorganic arsenic content in rice and its bioavailability. This study evaluated total arsenic and inorganic arsenic in rice cooked with arsenic-contaminated water, the bioaccessibility of As(III) and As(V) after simulated gastrointestinal digestion, and the extent of arsenic retention and transport by Caco-2 cells used as a model of intestinal epithelia. After cooking, inorganic arsenic contents increase significantly. After simulated gastrointestinal digestion, the bioaccessibility of inorganic arsenic reached 63 99%; As(V) was the main species found. In Caco-2 cells, arsenic retention, transport, and total uptake (retention + transport) varied between 0.6 and 6.4, 3.3 and 11.4, and 3.9 and 17.8%, respectively. These results show that in arsenic endemic areas with subsistence rice diets, the contribution of inorganic arsenic from cooked rice should be considered in assessments of arsenic health risk. PMID- 16248592 TI - Pesticides and watershed-scale modeling: solutions for water quality management. PMID- 16248593 TI - Comparison of regulatory estimates of drinking water concentrations with monitoring data. AB - Currently, regulatory practice in the United States is to estimate potential concentrations in drinking water from surface water by using an index reservoir scenario. This approach extrapolates results from the modeling of a single field with maximum application rates to the watershed scale, based on a percent crop area estimate. Since 1998, Bayer CropScience and its predecessor companies have conducted drinking water monitoring studies with a number of different compounds. The results from these studies show that the index reservoir scenario greatly overpredicts residues in surface water. The most important factor is the overestimation of use within a watershed. Other factors contributing to the overestimation of concentrations are the conservative procedures used to obtain the chemical fate related input parameters and the simplified hydrology. A new procedure based on the USGS WARP model, being developed by a group of scientists from the EPA, USGS, USDA, and industry, will provide more realistic estimates of concentrations of pesticides and their metabolites in drinking water. PMID- 16248594 TI - Comparison of regulatory method estimated drinking water exposure concentrations with monitoring results from surface drinking water supplies. AB - Crop-protection compounds are useful tools that enhance the quality of the food we enjoy. However, crop-protection products can enter aquatic systems either by direct or by indirect application. To better understand the possible frequency and magnitude of exposure to water resources, the regulatory community has developed a set of relatively straightforward models for estimating exposure to these water systems. The focus of this research was to compare how well the estimates of exposure to drinking water based on model calculations relate to actual monitoring data. Physical/chemical property data were entered in the EPA's exposure model FIRST and into PRZM/EXAMS. The predictions from FIRST and PRZM/EXAMS were then compared to actual monitoring data from a USGS/EPA cooperative program, which monitored for pesticides in vulnerable surface drinking water supplies during 1999 and 2000. Results from this examination indicate the exposure from the models can overpredict concentrations found in water by several orders of magnitude. An overprediction factor is presented that corrects model predictions to more closely approximate concentrations found in reservoirs (p = 0.05). PMID- 16248595 TI - Characterization of fate and transport of isoxaflutole, a soil-applied corn herbicide, in surface water using a watershed model. AB - The objective of this study is to conduct a comprehensive assessment of the transport of isoxaflutole and RPA 202248 and their accumulation potential in semistatic water bodies, using a distributed watershed scale model and observed water quality data. A conceptual model was developed to characterize the fate and transport of isoxaflutole residues to and in surface water. The soil and water assessment tool (SWAT), a continuous daily time-step watershed model, was used to simulate the processes identified in the conceptual model. Monitoring data were available for a number of surface water bodies within the major product use area as a result of extensive and intensive residue monitoring. Detailed product use information at the zip-code level was obtained through dealer sales and a grower survey. The hydrologic and chemical transport results from the SWAT model were validated by comparison to available monitoring data from selected water bodies. Upon validation, the model was used to simulate the fate of isoxaflutole-derived residues in the water bodies using long-term historical weather data. The results from this investigation indicate no evidence of long-term accumulation of isoxaflutole and its metabolite RPA 202248 in semistatic water bodies. PMID- 16248597 TI - Iron tris(dibenzoylmethane)-centered polylactide stars: multiple roles for the metal complex in lactide ring-opening polymerization. AB - Poly(lactic acid) (PLA)-supported dibenzoylmethane (dbm) and corresponding metal complexes have potential applications as biomaterials and catalysts. Using hydroxyl-functionalized dbm (i.e., dbmOH) as the initiator and Sn(oct)2 as the catalyst, lactide ring-opening polymerizations were unexpectedly slow (6 h), and PDIs broadened when molecular weights >10 000 were targeted. Because interactions between the dbm ligand and Sn(oct)2 may be responsible for the diminished catalyst activity and molecular weight control, iron(III) was employed as a protecting group to form Fe(dbmOH)3. Shorter reaction times (10 min) were noted with this trifunctional metalloinitiator and Sn(oct)2, and higher molecular weights were achievable. Moreover, it was discovered that Fe(dbmOH)3 serves not only as an initiator but also as a catalyst and activating group for the polymerization. Even without the tin catalyst, iron-centered polymers with low PDIs (<1.1) were obtained within 10 min ( approximately 70% monomer conversion). The resulting Fe(dbmPLA)3 stars were demetalated by acid treatment to generate dbmPLA for subsequent coordination to other metals. To explore the scope of iron beta-diketonate complexes as catalysts, additional studies were performed with dbmOH and benzyl alcohol initiators using either iron tris(dbm) or iron tris(acac) (acac = acetylacetonate) as the catalyst. Here too, PLA products were obtained, and the iron catalysts were readily separated by treatment with aqueous HCl. PMID- 16248596 TI - A strategy for the development of small-molecule-based sensors that strongly fluoresce when bound to a specific RNA. AB - We report a broadly applicable approach for the development of small-molecule based RNA sensors. Our photoinduced electron transfer (PET) sensor consists of a fluorescein derivative as the fluorophore and two aniline derivatives as electron donors (quenchers). The isolation of electron-donor-binding RNA by in vitro selection (also known as SELEX) yielded an RNA aptamer that could increase the fluorescence intensity of the sensor by 13-fold. This result shows that RNA electron-donor interactions can be used to develop modular RNA chemosensors. PMID- 16248598 TI - Direct observation of the disorder of the methyl group of (R)-1,2-diaminopropane ligand in the quasi-1D bromo-bridged Ni(III) complex by STM. AB - Scanning tunneling microscopy (STM) has been measured in quasi-one-dimensional (Quasi-ID) bromo-bridged Ni(III) complexes. In the STM image of [Ni(pn)2Br]Br2 (pn = (R)-1,2-diaminopropane), the bright spots are fluctuated zigzags. Such a result indicates that the methyl groups of the pn ligands are not arranged on the right- and left-hand alternatively along the chains, but in the domain structures or fluctuated structures. This is the first direct observation of the disorder of the methyl group of pn ligands in real space. PMID- 16248599 TI - Intermediates trapped during nitrogenase reduction of N triple bond N, CH3-N=NH, and H2N-NH2. AB - A high population intermediate has been trapped on the nitrogenase active site FeMo cofactor during reduction of N2. In addition, intermediates have been trapped during reduction of CH3-N=NH by the alpha-195Gln variant and during reduction of H2N-NH2 by the alpha-70Ala/alpha-195Gln variant. Each of these trapped states shows an EPR signal arising from an S = 1/2 state of the FeMo cofactor. 15N ENDOR shows that each intermediate has a nitrogenous species bound to the FeMo cofactor, with a single type of N seen for each bound intermediate. The g tensors are unique to each intermediate, g(e) = [2.084, 1.993, 1.969], g(m) = [2.083, 2.021, 1.993], g(l) = [2.082, 2.015, 1.987], as are the 15N hyperfine couplings at g1, which suggests that three distinct stages of NN reduction may have been trapped. The 1H ENDOR spectra show that the N2 intermediate is at a distinct and earlier stage of reduction from the other two, so at least two stages of NN reduction have been trapped. Some possible structures of the hydrazine intermediate are presented. PMID- 16248600 TI - A ratiometric fluorescence probe for selective visual sensing of Zn2+. AB - A simple ratiometric fluorescence probe based on vinylpyrrole end-capped bipyridine for the visual sensing of Zn2+ under aqueous physiological pH (6.8 7.4) is described. The fluorophores 3a-c showed strong emission around 537 nm in acetonitrile with a quantum yield of 0.4. In buffered (HEPES, pH 7.2) acetonitrile-water mixture (9:1 v/v), titration of transition metal salts to 3c showed strong quenching of the emission at 547 nm except in the case of Zn2+, which resulted in a red-shifted emission at 637 nm. Alkali and alkaline earth metal salts could not induce any considerable changes to the emission behavior of 3a-c. The binding of Zn2+ was highly selective in the presence of a variety of other metal ions. Though Cu2+ quenches the emission of 3c, in the presence of Zn2+, a red emission prevails, indicating the preference of 3c toward Zn2+. Job plot and Benesi-Hildebrand analysis revealed a 1:1 complexation between the probe and the metal ion. The selective visual sensing of Zn2+ with a red emission is ideally suited for the imaging of biological specimens. PMID- 16248601 TI - Diode or tunnel-diode characteristics? Resolving the catalytic consequences of proton coupled electron transfer in a multi-centered oxidoreductase. AB - Protein film voltammetry has been employed to define multiple catalytic consequences of proton coupled electron transfer (PCET) in a cytochrome c nitrite reductase. Current-potential profiles reflecting the steady-state rate of nitrite limited reduction have been defined from pH 4 to 8. Lowering the electrode potential at pH 8 causes the catalytic current to increase and then decrease before it takes a value independent of any further lowering of electrode potential. By comparison, at pH 4, catalysis is initiated at more positive electrode potentials in an approximately sigmoidal fashion with no attenuation of the catalytic rate evident at more negative electrode potentials. The results show that activity is turned on by the coupled transfer of two electrons and one proton to the enzyme. The decreased rate of catalysis at lower electrode potentials under more alkaline conditions shows that this rate attenuation occurs only when reduction is not coupled to compensating protonation(s) of the enzyme. Sites within the enzyme whose reduction and/or protonation may contribute to the definition of these activities are discussed. PMID- 16248602 TI - Assembly of polymeric silver(I) complexes of isomeric phenylenediethynides with the supramolecular synthons Agn subset C2-R-C2 superset Agn (R = p-, m-, o-C6H4; n = 4, 5). AB - New Agn subset C2-R-C2 supersetAgn (R = p-, m-, o-C6H4; n = 4, 5) supramolecular synthons have been explored in the coordination network assembly of silver(I) complexes of the isomeric phenylenediethynides. An unprecedented mu5-eta1 coordination mode for the ethynide moiety and a mixed mu4,mu5-coordination mode for the o-phenylenediethynide group are observed, providing a rationale for the abundant occurrence of C2@Agn (n 98% conversion in undistilled commercial grade toluene in the presence of 2 mol % of an air-stable Cu salt (CuCN) and a readily available chiral ligand. Enantioselective ACA reactions deliver products that can be readily functionalized to afford a variety of synthetically versatile compounds in high optical purity. PMID- 16248614 TI - One-dimensional assembly of chalcogenide nanoclusters with bifunctional covalent linkers. AB - Even though different approaches have been developed to achieve various 1D assemblies of nanocrystals, few studies have been done on the assembly of crystallographically well-defined chalcogenide nanoclusters. Here, by using bifunctional organic ligands as the directional linker, a series of one dimensional assemblies of semiconducting chalcogenide nanoclusters have been prepared and characterized. The synthetic method allows for the preparation of differently sized tetrahedral nanoclusters that are joined together with organic linkers of different length and rigidity. Multiple linking modes between nanoclusters and organic ligands are revealed in four different assemblies that also exhibit size-dependent optical properties. PMID- 16248615 TI - Nanosphere arrays with controlled sub-10-nm gaps as surface-enhanced raman spectroscopy substrates. AB - We demonstrate a convenient and cost-effective chemical approach for fabricating highly ordered Au nanoparticle arrays with sub-10-nm interparticle gaps. Near field enhancements inside the interparticle gaps create uniform periodic arrays of well-defined "hot spots" exploitable for large surface-enhanced Raman spectroscopy (SERS) enhancements. A cetyltrimethylammonium bromide (CTAB) bilayer surrounding each individual nanoparticle upon array crystallization is responsible for this periodic gap structure; displacement of the CTAB by smaller thiolated molecules does not affect the structural integrity of the arrays. As SERS substrates, the as-fabricated Au nanoparticle arrays exhibit high SERS sensitivity, long-term stability, and consistent reproducibility. PMID- 16248616 TI - Relaying asymmetry of transient atropisomers of o-iodoanilides by radical cyclizations. AB - Atropisomers of N-2 degrees -alkyl-N-acryloyl-2-iodoanlides have been resolved by chromatography and crystallization-induced asymmetric transformation. These molecules have atropisomerization barriers of 23-24 kcal/mol and return to equilibrium ratios over several hours at ambient temperature in solution. The transient chirality can be locked in by radical cyclizations, which provide N-2 degrees -alkyl-3-methyl-1,3-dihydroindol-2-ones with high levels of chirality transfer. The mechanistic model features a stereoselective aryl radical cyclization that is more rapid than the rotation of the N-aryl bond of the anilide. PMID- 16248617 TI - n-type organic field-effect transistors with very high electron mobility based on thiazole oligomers with trifluoromethylphenyl groups. AB - Novel thiazole oligomers and thiazole/thiophene co-oligomers with trifluoromethylphenyl groups were developed as n-type semiconductors for OFETs. They showed excellent n-type performances with high electron mobilities. A 5,5' bithiazole with trifluoromethylphenyl groups forms a closely packed two dimensional columnar structure leading to a high performance n-type FET. The electron mobility was enhanced to 1.83 cm2/Vs on the OTS-treated substrate. PMID- 16248618 TI - The rotational spectrum and structure of HOOOH. AB - Dihydrogen trioxide, HOOOH, which is a species with fundamental importance for understanding the chain formation ability of the oxygen atom, was detected in a supersonic jet by a Fourier transform microwave spectrometer with a pulsed discharge nozzle, together with double resonance and triple resonance techniques. Its precise molecular structure was determined from the experimentally determined rotational constants of HOOOH and its isotopomer, DOOOD. Many of the microwave and millimeter wave transitions can now be accurately predicted, which could be facilitated for remote sensing of the molecule to elucidate its roles in various chemical processes. PMID- 16248619 TI - In(OH)BDC.0.75BDCH2 (BDC = Benzenedicarboxylate), a hybrid inorganic-organic vernier structure. AB - The compound In(OH)BDC.0.75BDCH2 (BDC = benzenedicarboxylate), 1, has been synthesized and characterized by single-crystal X-ray diffraction. The structure comprises two distinct sublattices formed by a covalently linked In(OH)BDC lattice and ordered chains of hydrogen-bonded H2BDC molecules and can be described as a hybrid inorganic coordination polymer-organic vernier structure. Each InO6 octahedron of the octahedral chain has a length of 3.6 A along the chain axis, whereas each H2BDC molecule has a length of 9.6 A along the guest column axis. Therefore, a unit of eight InO6 octahedra of the octahedral chain is just in registry with three H2BDC molecules of the guest column giving a repeat unit of 28.76 A along the channel axis direction. PMID- 16248621 TI - DNA damage by fasicularin. AB - Fasicularin is a structurally novel thiocyanate-containing alkaloid isolated from the ascidian Nephteis fasicularis. Early biological experiments suggested that this compound's cytotoxic properties may stem from its ability to damage cellular DNA. Sequence gel analysis reveals that treatment of a 5'-32P-labeled DNA duplex with fasicularin in pH 7.0 buffer causes strand cleavage selectively at guanine residues. Further experiments indicate that production of these base-labile lesions in DNA involves alkylation of guanine residues by a fasicularin-derived aziridinium ion. This work reveals fasicularin as the first natural product found to generate a DNA-alkylating aziridinium ion via a mechanism analogous to the clinically used anticancer drugs mechlorethamine, melphalan, and chlorambucil. PMID- 16248620 TI - Isochorismate pyruvate lyase: a pericyclic reaction mechanism? AB - Isochorismate pyruvate lyase (IPL) catalyzes the cleavage of isochorismate to give salicylate and pyruvate, a key step in bacterial siderophore biosynthesis. We investigated the enzyme from Pseudomonas aeruginosa using isochorismate selectively deuterated at C2 as a substrate. Monitoring the reaction by 2H NMR spectroscopy revealed that the label is quantitatively transferred from C2 to C9, producing stoichiometric amounts of [3-2H]pyruvate as product. Moreover, the deuterium kinetic isotope effect of 2.34 +/- 0.08 on kcat indicates that C-H cleavage is significantly rate limiting. Consistent with these data, hybrid density functional theory (HDFT) calculations at the Becke3LYP/DZ+(2d,p) level of theory predict a concerted but highly asynchronous pericyclic transition structure, in which carbon-oxygen bond cleavage is more advanced than hydrogen atom transfer from C2 to C9; the calculated 2H isotope effect of 2.22 at C2 is in excellent accord with the experimental value. Together, these findings indicate that IPL should be added to the small set of proteins that are known to catalyze pericyclic reactions. They also raise the possibility that enzymes, such as chorismate pyruvate lyase, salicylate synthase, 4-amino-4-deoxychorismate lyase, and anthranilate synthase, which accelerate formally similar reaction steps, may also exploit pericyclic mechanisms. PMID- 16248622 TI - 3/2-order chain kinetics involving a postulated dicationic intermediate in the isomerization of a p-isocyano to a p-cyano azaphosphatrane monocation. AB - Kinetic evidence suggests the possibility of a dicationic intermediate in the title reaction. Thus the linkage isomerization reaction, PNC+ = PCN+, is described by the rate law, nu = 3/2k[PNC+]3/2, which can be interpreted by a chain mechanism with the propagation reaction PNC+ + P2+ --> P2+ + PCN+. Such propagation is unusual in that the intermediate regenerates itself in this single step rather than forming a different intermediate for a second propagation step. Cyanide ions inhibit the rate because they participate in the termination step, P2+ + CN- --> PCN+. The rate constant in CD3CN at 100 degrees C is 3/2k = 7.2 +/- 0.6 x 10-5 L1/2 mol-1/2 s-1; 3/2k represents the composite (kinit/kterm)1/2 kprop. When the reaction is carried out in the presence of PBr+, however, the reaction becomes much faster and is described by the rate law, nu = kBr[PBr+][PNC+]; because [PBr+] remains at constant concentration, the time course experiments follow first-order kinetics. PMID- 16248623 TI - ZiCo: a peptide designed to switch folded state upon binding zinc. AB - We describe a novel approach to the design of a metal-triggered conformational switch. Specifically, two distinct protein-folding motifs were merged into one polypeptide sequence. The target structures were an alpha-helical coiled-coil trimer and zinc-bound monomer. Solution-phase spectroscopic, sedimentation, and binding studies confirmed the key aspects of the design. Both forms of the peptide were cooperatively folded, and the switch between them was reversible. This design process potentially presents a novel route to peptide-based biosensors. PMID- 16248624 TI - Synthesis, characterization, and structure of cyclopenta[c]thiophenes and their manganese complexes. AB - 1,3-Diaryl-4H-cyclopenta[c]thiophenes are efficiently prepared from 1,2 diaroylcyclopentadienes by use of Lawesson's reagent. eta5-Cyclopenta[c]thienyl complexes, [Mn(eta5-SC7H3-1,3-R2)(CO)3] (R = Me, Ph), are prepared in high yield by ligand substitution reactions of [MnBr(CO)5] with [SnMe3(SC7H3-1,3-R2)]. Alternatively, thiation with P4S10/NaHCO3 converts [Mn{eta5-1,2-C5H3(COR)2)(CO)3] to [Mn(eta5-SC7H3-1,3-R2)(CO)3] (R = Ph, 4-tolyl, 4-MeOC6H4, benzo[2,3 b]thienyl). The molecular structures of complexes with R = Me, Ph show planar eta5-cyclopenta[c]thienyl ligands, with the manganese atom slightly displaced away from the ring-fusion bond. PMID- 16248625 TI - Oxidation reactions of the phosphinidene oxide ligand. AB - The (H-DBU)+ salt of the anionic phosphinidene oxide complex [MoCp(CO)2{P(O)R*}]- (1) (DBU = 1,8-diazabicyclo[5.4.0]undec-7-ene; R* = 2,4,6-C6H2tBu3) reacts with different oxidizing agents, displaying a multisite activity located at the Mo and P atoms or at the Mo=P bond. Thus, reaction of 1 with [FeCp2]BF4 gives the dimer [Mo2Cp2(CO)4{P(O)R*}2], and reaction with bromine gives the phosphinous acid complex [MoBrCp{P(OH)(CH2CMe2C6H2tBu2}(CO)2], the latter arising from an unprecedented C-H bond addition to the oxide P=O moiety. In contrast, reaction of 1 with p-benzoquinone occurs at the P site to give the P,O-bound phosphonite complex [MoCp{kappa2-OP(OC6H4OH)R*}(CO)2]. Finally, oxygen or sulfur atoms are added to the Mo=P bond by reaction of 1 with Me2CO2 and S8 to give the novel dioxophosphorane or thiooxophosphorane complexes [MoCp(CO)2{kappa2-EP(O)R*}]- (E = O, S). The thiooxophosphorane anion is a good nucleophile and is methylated at either the S or O positions depending on the electrophile used (MeI or (Me3O)BF4) to give the isomers [MoCp{kappa2-(MeS)P(O)R*}(CO)2] and [MoCp{kappa2 SP(OMe)R*}(CO)2], both having novel organophosphorus ligands. PMID- 16248626 TI - EPR distance measurements support a model for long-range radical initiation in E. coli ribonucleotide reductase. AB - The class I E. coli ribonucleotide reductase, composed of homodimers of R1 and R2, catalyzes the conversion of nucleoside diphosphates to deoxynucleoside diphosphates. The reduction process involves the tyrosyl radical on R2 that generates a transient thiyl radical on R1 over a proposed distance of 35 A. A mechanism-based inhibitor, 2'-azido-2'-deoxyuridine-5'-diphosphate, that reduces the tyrosyl radical on R2 and forms a nitrogen-centered radical on R1 has provided a method to measure the diagonal distance between the two subunits. PELDOR and DQC paramagnetic resonance methods give rise to a distance of 48 A, similar to that calculated from a docking model of the R1 and R2 structures. PMID- 16248627 TI - Highly stereoselective [2,3]-sigmatropic rearrangement of sulfur ylide generated through Cu(I) carbene and sulfides. AB - A highly stereoselective [2,3]-sigmatropic rearrangement of sulfur ylide generated through Cu(I) carbene and allyl and propargyl sulfides by a double asymmetric induction approach that combines a chiral camphor sultam auxiliary and Cu(I) catalyst with chiral or achiral diimine ligands has been developed. PMID- 16248628 TI - Proton-mediated electron configuration change in high-spin iron(II) porphyrinates. AB - The synthesis, molecular structure, and electronic structure characterization of two five-coordinate high-spin imidazolate-ligated iron(II) porphyrinates are reported. Their electronic structure, as deduced from Mossbauer spectra obtained in strong magnetic fields, is distinctly different from that of the analogous imidazole-ligated species. The resulting electronic structure models are consistent with all observed differing features in the two classes. PMID- 16248629 TI - A simple route to difluorocarbene and perfluoroalkylidene complexes of iridium. AB - Primary perfluoroalkyl compounds of iridium undergo facile two-electron reduction to afford a simple route to difluorocarbene and perfluoroalkylidene complexes, two of which have been crystallographically characterized. Low-temperature protonation reactions illustrate that the thermodynamic site of protonation depends on the nature of the fluorinated carbene; the difluorocarbene complex undergoes protonation at iridium, while its perfluoroethylidene congener undergoes protonation at carbon. PMID- 16248630 TI - Synthesis of 2,3-disubstituted benzofurans by platinum-olefin-catalyzed carboalkoxylation of o-alkynylphenyl acetals. AB - The PtCl2-catalyzed cyclization reaction of o-alkynylphenyl acetals 1 in the presence of 1,5-cyclooctadiene produces 3-(alpha-alkoxyalkyl)benzofurans 2 in good to high yields. For example, the reaction of acetaldehyde ethyl 2-(1 octynyl)phenyl acetal (1a), acetaldehyde ethyl 2-(cyclohexylethynyl)phenyl acetal (1c), and acetaldehyde ethyl 2-(phenylethynyl)phenyl acetal (1f) in the presence of 2 mol % of platinum(II) chloride and 8 mol % of 1,5-cyclooctadiene in toluene at 30 degrees C gave the corresponding 2,3-disubstituted benzofurans 2a, 2c, and 2f in 91, 94, and 88% yields, respectively. PMID- 16248631 TI - Heterocycles by PtCl2-catalyzed intramolecular carboalkoxylation or carboamination of alkynes. AB - PtCl2 constitutes a convenient catalyst for intramolecular hydroalkoxylation, carboalkoxylation, hydroamination, and carboamination reactions of alkynes, effecting the formation of substituted benzo[b]furan, indole-, and isochromene-1 one derivatives, respectively. This procedure allows for the transfer of (substituted) allyl, methoxymethyl (MOM), benzyloxymethyl (BOM), and (trimethylsilyl)ethoxymethyl (SEM) groups from oxygen to carbon and is compatible with functional groups that are susceptible to oxidative insertion of low valent metal species previously used for similar purposes. Although some of the reactions can even be carried out in air, the rates are significantly increased when conducted under an atmosphere of CO. A mechanistic rationale is proposed, implying activation of the alkyne by the carbophilic Pt(2+) template as the primary step of the catalytic cycle. PMID- 16248632 TI - Total synthesis of reblastatin. AB - Enantioselective total synthesis of reblastatin is described. The synthesis highlights hydrozirconation, transmetalation, aldehyde addition sequence to install E-trisubstituted olefin and C7 stereocenter, and the first use of an intramolecular Buchwald-like amidation reaction to close the 19-membered macrolactam. PMID- 16248633 TI - Synthesis of dihydronaphthalenes via aryne Diels-Alder reactions: scope and diastereoselectivity. AB - Novel aryne Diels-Alder reactions with functionalized acyclic dienes are reported. These give useful cis-substituted dihydronaphthalene building blocks in good yield which are not easily accessible via other means, as demonstrated in the synthesis of sertraline. The first asymmetric aryne cycloaddition with an acyclic diene is also reported, giving excellent diastereoselectivities with Oppolzer's sultam as a chiral auxiliary. PMID- 16248634 TI - Controlled fabrication of 1D molecular lines across the dimer rows on the Si(100) (2 x 1)-H surface through the radical chain reaction. AB - Current interest in the fabrication of organic nanostructures on silicon surface is focused on the self-directed growth of 1D molecular lines with predefined position, structure, composition, and the length on the H-terminated Si(100)-(2 x 1) surface. To date, no studies have succeeded in growing the molecular line across the dimer rows on Si(100)-(2 x 1)-H, which is highly desirable. Using scanning tunneling microscopy (STM), we studied a new molecular system (allyl mercaptan, CH2=CH-CH2-SH) that undergoes chain reaction across the dimer row on the Si(100)-(2 x 1)-H surface at 300 K and produces covalently bonded 1D molecular lines. In combination with the previous findings of chain reaction along the rows, the present observations of self-directed growth of 1D molecular lines across the dimer rows on the Si(100)-(2 x 1)-H surface provide a means to connect any two points (through molecular lines) on a 2D surface. PMID- 16248635 TI - Composite alignment media for the measurement of independent sets of NMR residual dipolar couplings. AB - The measurement of independent sets of NMR residual dipolar couplings (RDCs) in multiple alignment media can provide a detailed view of biomolecular structure and dynamics, yet remains experimentally challenging. It is demonstrated here that independent sets of RDCs can be measured for ubiquitin using just a single alignment medium composed of aligned bacteriophage Pf1 particles embedded in a strained polyacrylamide gel matrix. Using this composite medium, molecular alignment can be modulated by varying the angle between the directors of ordering for the Pf1 and strained gel matrix, or by varying the ionic strength or concentration of the Pf1 particles. This approach offers significant advantages in that greater experimental control can be exercised over the acquisition of multi-alignment RDC data while a homogeneous chemical environment is maintained across all of the measured RDC data. PMID- 16248636 TI - Single crystal manganese oxide multipods by oriented attachment. AB - Nonhydrolytic sol-gel processes in organic solvents have become very popular for the synthesis of metal oxide NCs. We report an affordable, high-yield, shape control synthesis of MnO multipod nanocrystals with a quick reaction time. The reaction yields exclusively multipods; two to six pod nanocrystals are synthesized. The mechanism leading to this hierarchical nanostructure is studied in relation with an oriented attachment mechanism; all nanocrystals are found to be single crystals. The study is completed by high-resolution TEM, X-ray powder diffraction, and magnetic measurements. PMID- 16248637 TI - Catalytic asymmetric reductive Michael cyclization. AB - A highly efficient and chemo-, regio-, diastereo-, and enantioselective organocatalytic tandem conjugate reduction-Michael cyclization of enal enones has been developed. Accordingly, treating the enal enone with a Hantzsch dihydropyridine in the presence of a catalytic amount of an imidazolidinone organocatalyst provides cyclic keto aldehydes in high yields and enantiomeric excesses. The reaction works well with aliphatic and aromatic substrates in the synthesis of five- and six-membered carbacyclic derivatives. PMID- 16248638 TI - Stereocontrolled total synthesis of (-)-eudistomin C. AB - A stereocontrolled total synthesis of (-)-eudistomin C was accomplished in 18 step sequence with an overall yield of 7.7%. The synthesis features the diastereoselective Pictet-Spengler reaction of a tryptamine derivative and the Garner aldehyde catalyzed by Bronsted acids, and the unprecedented construction of the unusual oxathiazepine ring by intramolecular alkylation. PMID- 16248639 TI - Direct experimental observation of the effect of the base pairing on the oxidation potential of Guanine. AB - The effect of complementary base pairing on the oxidation potential of a guanosine derivative has been determined by cyclic and differential pulse voltammetry in CHCl3. The formation of the Watson-Crick H-bonded complex lowers the oxidation potential of the free molecule by 0.34 V, which compares well with the value obtained by DFT/B3LYP/6-311++g** computations. PMID- 16248641 TI - Directing and sensing changes in molecular conformation on individual carbon nanotube field effect transistors. AB - This study explores how to populate the surface of the carbon nanotubes with functional molecules that can be toggled back-and-forth between different molecular conformations. The molecules synthesized for this study are tagged with a photoswitchable headgroup and a functional group which directs the assembly on the surface of the carbon nanotubes. Single-walled carbon nanotube field effect transistors switch between high and low conductance as the molecules are switched with light between open and closed conformations. These devices detect the photoswitching of approximately 104 molecules. PMID- 16248640 TI - Discovery of a biologically active thiostrepton fragment. AB - Design, synthesis, and biological evaluation of several domains of the thiopeptide antibiotic thiostrepton led to the discovery of a biologically active fragment. The biological properties of this novel small organic molecule include antibiotic activity against methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus faecalis (VREF) bacterial strains, as well as cytotoxic action against a number of cancer cell lines. PMID- 16248642 TI - An unprecedented twist to ODCase catalytic activity. AB - Orotidine-5'-monophosphate decarboxylase (ODCase) has evolved to catalyze a decarboxylation reaction, most probably via a carbanion species at the C6 position of orotidine-5'-monophosphate. We reveal an unusual biochemical pathway of conversion of 6-cyano-uridine-5'-monophosphate by ODCase to barbiturate-5' monophosphate via perhaps an electrophilic center at the C6 position, leading to inhibition. This potential of ODCase is very useful in the design of novel inhibitors. PMID- 16248643 TI - Enantioselective organo-cascade catalysis. AB - A new strategy for organocatalysis based on the biochemical blueprints of biosynthesis has enabled a new laboratory approach to cascade catalysis. Imidazolidinone-based catalytic cycles, involving iminium and enamine activation, have been successfully combined to allow a large diversity of nucleophiles (furans, thiophenes, indoles, butenolides, hydride sources, tertiary amino lactone equivalents) and electrophiles (fluorinating and chlorinating reagents) to undergo sequential addition with a wide array of alpha,beta-unsaturated aldehydes. These new cascade catalysis protocols allow the invention of enantioselective transformations that were previously unknown, including the asymmetric catalytic addition of the elements of HF across a trisubstituted olefin. Importantly, these domino catalysis protocols can be mediated by a single imidazolidinone catalyst or using cycle-specific amine catalysts. In the latter case, cascade catalysis pathways can be readily modulated to provide a required diastereo- and enantioselective outcome via the judicious selection of the enantiomeric series of the amine catalysts. A central benefit of combining multiple asymmetric organocatalytic events into one sequence is the intrinsic requirement for enantioenrichment in the second induction cycle, as demonstrated by the enantioselectivities obtained throughout this study (>/=99% ee in all cases). PMID- 16248644 TI - Periodic ab initio approach for the cooperative effect of CH/pi interaction in crystals: relative energy of CH/pi and hydrogen-bonding interactions. AB - The bonding property of the CH/pi interaction in organic crystals has been investigated by the means of a periodic ab initio method. The energy of the CH(sp(2))/pi interaction in crystals, estimated with periodic RHF/6-21G*, showed a reasonable attractive CH(sp(2))/pi interaction owing to a cooperative effect, whereas the results calculated with RHF/cc-pVDZ indicate a negligibly small or repulsive interaction. The relative contribution of the CH(sp(2))/pi interaction to the column packing energy was found to be roughly half of the energy of a conventional hydrogen bond. The calculation of the charge distributions on the aromatic rings participating in the CH(sp(2))/pi interaction in crystals revealed that the atoms were more ionic than those in the gas phase. These theoretical calculations suggest a hydrogen-bonding characteristic for the CH(sp(2))/pi interaction in crystals, which does not occur in solution nor gas phase. We present computational evidence of the existence of the cooperative effect of CH(sp(2))/pi interaction in crystals. PMID- 16248645 TI - Catalytic transesterification of dialkyl phosphates by a bioinspired dicopper(II) macrocyclic complex. AB - For a number of phosphoryltransfer enzymes, including the exonuclease subunit of DNA polymerase I, a mechanism involving two-metal ions and double Lewis-acid activation of the substrate, combined with leaving group stabilization, has been proposed. Inspired by the active site structure of this enzyme, we have designed as a synthetic phosphoryl transfer catalyst the dicopper(II) macrocyclic complex LCu(2). Crystal structures of complexes [(L)Cu(2)(mu-NO(3))(NO(3))](NO(3))(2) (1), [(L)Cu(2)(mu-CO(3))(CH(3)OH)](BF(4))(2) (2), and [(L)Cu(2)(mu O(2)P(OCH(3))(2))(NO(3))](NO(3))(2) (3) illustrate various possibilities for the interaction of oxoanions with the dicopper(II) site. 1 efficiently promotes the transesterification of dimethyl phosphate (DMP) in CD(3)OD, k(cat) = 2 x 10(-)(4) s(-)(1) at 55 degrees C. 1 is the only available catalyst for the smooth transesterification of highly inert simple dialkyl phosphates. From photometric titrations and the pH dependence of reactivity, we conclude that a complex [(L)Cu(2)(DMP)(OCH(3))](2+) is the reactive species. Steric bulk at the -OR substituents of phosphodiester substrates O(2)P(OR)(2)(-) drastically reduces the reactivity of 1. This is explained with -OR leaving group stabilization by Cu coordination, an interaction which is sensitive to steric crowding at the alpha-C atom of substituent R. A proposed reaction mechanism related to that of the exonuclease unit of DNA polymerase I is supported by DFT calculations on reaction intermediates. The complex [(L)Cu(3)(mu(3)-OH)(mu CH(3)O)(2)(CH(3)CN)(2)](ClO(4))(3) (4) incorporates a [Cu(OH)(OCH(3))(2)(CH(3)CN)(2)](-) complex anion, which might be considered as an analogue of the [PO(2)(OCH(3))(2)(OCD(3))](2)(-) transition state (or intermediate) of DMP transesterification catalyzed by LCu(2). PMID- 16248646 TI - A versatile toolbox for variable DNA functionalization at high density. AB - To broaden the applicability of chemically modified DNAs in nano- and biotechnology, material science, sensor development, and molecular recognition, strategies are required for introducing a large variety of different modifications into the same nucleic acid sequence at once. Here, we investigate the scope and limits for obtaining functionalized dsDNA by primer extension and PCR, using a broad variety of chemically modified deoxynucleotide triphosphates (dNTPs), DNA polymerases, and templates. All natural nucleobases in each strand were substituted with up to four different base-modified analogues. We studied the sequence dependence of enzymatic amplification to yield high-density functionalized DNA (fDNA) from modified dNTPs, and of fDNA templates, and found that GC-rich sequences are amplified with decreased efficiency as compared to AT rich ones. There is also a strong dependence on the polymerase used. While family A polymerases generally performed poorly on "demanding" templates containing consecutive stretches of a particular base, family B polymerases were better suited for this purpose, in particular Pwo and Vent (exo-) DNA polymerase. A systematic analysis of fDNAs modified at increasing densities by CD spectroscopy revealed that single modified bases do not alter the overall B-type DNA structure, regardless of their chemical nature. A density of three modified bases induces conformational changes in the double helix, reflected by an inversion of the CD spectra. Our study provides a basis for establishing a generally applicable toolbox of enzymes, templates, and monomers for generating high density functionalized DNAs for a broad range of applications. PMID- 16248647 TI - Mechanistic insights into the phosphine-free RuCp*-diamine-catalyzed hydrogenation of aryl ketones: experimental and theoretical evidence for an alcohol-mediated dihydrogen activation. AB - The commercially available chiral diamine quincorine-amine, originally derived from quinine, was found to be a highly active catalyst for ruthenium-catalyzed hydrogenation of ketones. The complex formed between the quincorine-amine, containing both a primary and a quinuclidine amino function, and RuCp*Cl catalyzes the hydrogenation of aromatic and aliphatic ketones in up to 90% ee approximately 24 times faster than previously reported Ru-diamine complexes. The pseudo-enantiomer of the quincorine-amine, i.e., quincoridine-amine, also showed high activity; however, the enantioselectivities obtained with this catalyst were lower. The reason for the lower, but opposite stereoselectivity seen with the quincoridine-amine, as compared to the quincorine-amine, was rationalized by a kinetic and computational study of the mechanism of the reaction. The theoretical calculations also revealed a significantly lower activation barrier for the alcohol-mediated split of dihydrogen, as compared to the nonalcohol-mediated process, a finding of utmost implication also for the diphosphine/diamine mediated enantioselective hydrogenation of ketones. PMID- 16248648 TI - Electronic and steric effects on molecular dihydrogen activation in [CpOsH4(L)]+ (L = PPh3, AsPh3, and PCy3). AB - Single-crystal neutron diffraction, inelastic neutron scattering, and density functional calculations provide experimental and theoretical analyses of the nature of the osmium-bound, "elongated" dihydrogen ligands in [Cp*OsH(4)(L)][BF(4)] complexes (L = PPh(3), AsPh(3), or PCy(3)). The PPh(3) and AsPh(3) complexes clearly contain one dihydrogen ligand and two terminal hydrides; the H(2) ligand is transoid to the Lewis base, and the H-H vector connecting the central two hydrogen atoms lies parallel to the Ct-Os-L plane (Ct = centroid of Cp* ring). In contrast, in the PCy(3) complex the H-H vector is perpendicular to the Ct-Os-L plane. Not only the orientation of the central two hydrogen atoms but also the H-H bond length between them depends significantly on the nature of L: the H...H distance determined from neutron diffraction is 1.01(1) and 1.08(1) A for L = PPh(3) and AsPh(3), respectively, but 1.31(3) A for L = PCy(3). Density functional calculations show that there is a delicate balance of electronic and steric influences created by the L ligand that change the molecular geometry (steric interactions between the Cp* and L groups most importantly change the Ct-Os-L angle), changing the relative energy of the Os 5d orbitals, which in turn govern the H-H distance, preferred H-H orientation, and rotational dynamics of the elongated dihydrogen ligand. The geometry of the dihydrogen ligand is further tuned by interactions with the BF(4)(-) counterion. The rotational barrier of the bound H(2) ligand in [Cp*OsH(4)(PPh(3))](+), determined experimentally (3.1 kcal mol(-)(1)) from inelastic neutron scattering experiments, is in reasonable agreement with the B3LYP calculated H(2) rotational barrier (2.5 kcal mol(-)(1)). PMID- 16248649 TI - Knots in globule and coil phases of a model polyethylene. AB - We examine the statistics of knots with numerical simulations of a simplified model of polyethylene. We can simulate polymers of up to 1000 monomers (each representing roughly three CH(2) groups), at a range of temperatures spanning coil (good solvent) and globule (bad solvent) phases. We quantify the abundance of knots in the globule phase and in confined polymers, and their rarity in the swollen phase. Since our polymers are open, we consider (and test) various operational definitions for knots, which are rigorously defined only for closed chains. We also associate a typical size with individual knots, which are found to be small (tight and localized) in the swollen phase but large (loose and spread out) in the dense phases. PMID- 16248650 TI - Soft vesicles formed by diblock codendrimers of poly(benzyl ether) and poly(methallyl dichloride). AB - The synthesis of a block codendrimer (g3-PBE-b-g3-PMDC), composed of a third generation poly(benzyl ether) (PBE) monodendron and an aliphatic polyether (PMDC) monodendron is reported. In THF/diiospropyl ether (1:1) the PMDC block functions as a "hydrophilic" block, while the PBE acts as a "hydrophobic" block. The codendrimer can form interdigitated layers leading to vesicle formation. Tapping mode atomic force microscopy (AFM), dynamic light scattering (DLS), and transmission electron microscopy (TEM) were used to characterize the vesicles. The effect of molecular architecture on the formation of the interdigitated layers and vesicles was studied. PMID- 16248651 TI - Transport of organic solutes through amorphous teflon AF films. AB - Fluorous media have great potential for selective extraction (e.g., as applied to organic synthesis). Fluorous polymer films would have significant advantages in fluorous separations. Stable films of Teflon AF 2400 were cast from solution. Films appear defect-free (SEM; AFM). Rigid aromatic solutes are transported (from chloroform solution to chloroform receiving phase) in a size-dependent manner (log permeability is proportional to -0.0067 times critical volume). Benzene's permeability is about 2 orders of magnitude higher than in comparable gas-phase experiments. The films show selectivity for fluorinated solutes in comparison to the hydrogen-containing control. Transport rates are dependent on the solvent making up the source and receiving phases. The effect of solvent is, interestingly, not due to changes in partition ratio, but rather it is due to changes in the solute diffusion coefficient in the film. Solvents plasticize the films. A less volatile compound, -COOH-terminated poly(hexafluoropropylene oxide) (4), plasticizes the films (T(g) = -40 degrees C). Permeabilities are decreased in comparison to 4-free films apparently because of decreased diffusivity of solutes. The slope of dependence of log permeability on critical volume is not changed, however. PMID- 16248652 TI - Triplet state photosensitization of nanocrystalline metal oxide electrodes by zinc-substituted cytochrome c: application to hydrogen evolution. AB - The interfacing of nanostructured semiconductor photoelectrodes with redox proteins is an innovative approach to the development of artificial photosynthetic systems. In this paper, we have investigated the photoinduced electron-transfer reactions of zinc-substituted cytochrome c, ZnCyt-c, immobilized on mesoporous, nanocrystalline metal oxide electrodes. Efficient electron injection from the triplet state of ZnCyt-c is observed into TiO(2) electrodes (t(50%) approximately 100 micros) resulting in a long-lived charge separated state (lifetime of up to 0.4 s). Further studies were undertaken as a function of electrolyte pH and metal oxide employed. Optimum yield of a long lived charge-separated state was observed employing TiO(2) electrodes at pH 5, consistent with our previous studies of analogous dye-sensitized metal oxide electrodes. The addition of EDTA as a sacrificial electron donor to the electrolyte resulted in efficient photogeneration of molecular hydrogen, with a quantum yield per one absorbed photon of 10 +/- 5%. PMID- 16248653 TI - Common semiopen conformations of Mg2+-free Ras, Rho, Rab, Arf, and Ran proteins combined with GDP and their similarity with GEF-bound forms. AB - A computational study was performed on the Mg(2+)-free conformations of the small guanine nucleotide-binding proteins (GNBPs): Ras, Rho, Rab, Arf, and Ran, which were complexed with GDP. Molecular dynamics (MD) simulation was executed for each complex for the duration of 3.0 ns to investigate the effects of Mg(2+) ions on the GNBPs' structure. The results indicated that all Mg(2+)-free GNBPs formed a groove between the switch region and the nucleotide-binding site. In some GNBP families, the release of Mg(2+) was reported to play an important role in binding the guanine nucleotide-exchanging factor (GEF) promoting the GDP/GTP exchange reaction. Interestingly, the grooves, which appeared in the MD simulations, were similar to the grooves experimentally observed in the GNBP-GEF complex. We also calculated the Mg(2+)-bound GNBPs to compare with the Mg(2+)-free forms. No groove was observed in the Mg(2+)-bound GNBPs. These results demonstrated a regulatory role of Mg(2+) ion to prepare a template for the GEF binding. Moreover, the results suggested that the release of Mg(2+) ion lead to the GEF GNBP binding. PMID- 16248654 TI - Separating the contribution of translational and rotational diffusion to protein association. AB - The association of two proteins is preceded by a mutual diffusional search in solution. The role of translational and rotational diffusion in this process has been studied theoretically for many years. However, systematic experimental verification of theoretical results is still lacking. We report here measurements of association rates of the proteins beta-lactamase (TEM) and beta-lactamase inhibitor protein (BLIP) in solutions of glycerol and poly(ethylene glycol) of increasing viscosity. We also measured translational and rotational diffusion in the same solutions, using fluorescence correlation spectroscopy and fluorescence anisotropy, respectively. It is found that in glycerol both translational and rotational diffusion rates are inversely dependent on viscosity, as predicted by the classical Stokes-Einstein relations, while the association rate depends nonlinearly on viscosity. In contrast, the association rate depends only weakly on the viscosity of the polymer solutions, which results in a similar weak dependence of k(on) on viscosity. The data are modeled using the theory of diffusion-limited association. Deviations from the theory are explained by a short-range solute-induced repulsion between the proteins in glycerol solution and an attractive depletion interaction generated by the polymers. These results open the way to the creation of a unified framework for all nonspecific effects involved in the protein association process, as well as to better theoretical understanding of these effects. Further, they reflect on the complex factors controlling protein association within the crowded environment of cells and suggest that a high concentration of macromolecules does not significantly impede protein association. PMID- 16248655 TI - Understanding how the herpes thymidine kinase orchestrates optimal sugar and nucleobase conformations to accommodate its substrate at the active site: a chemical approach. AB - The herpes virus thymidine kinase (HSV-tk) is a critical enzyme for the activation of anti-HSV nucleosides. However, a successful therapeutic outcome depends not only on the activity of this enzyme but also on the ability of the compound(s) to interact effectively with cellular kinases and with the target viral or cellular DNA polymerases. Herein, we describe the synthesis and study of two nucleoside analogues built on a conformationally locked bicyclo[3.1.0]hexane template designed to investigate the conformational preferences of HSV-tk for the 2'-deoxyribose ring. Intimately associated with the conformation of the 2' deoxyribose ring is the value of the C-N torsion angle chi, which positions the nucleobase into two different domains (syn or anti). The often-conflicting sugar and nucleobase conformational parameters were studied using North and South methanocarbadeoxythymidine analogues (6 and 7), which forced HSV-tk to make a clear choice in the conformation of the substrate. The results provide new insights into the mechanism of action of this enzyme, which cannot be gleaned from a static X-ray crystal structure. PMID- 16248656 TI - Evidence of a borderline region between E1cb and E2 elimination reaction mechanisms: a combined experimental and theoretical study of systems activated by the pyridine ring. AB - We report a combined experimental and theoretical study to characterize the mechanism of base-induced beta-elimination reactions in systems activated by the pyridyl ring, with halogen leaving groups. The systems investigated represent borderline cases, where it is uncertain whether the reaction proceeds via a carbanion intermediate (E1cb, A(xh)D(H) + D(N)) or via the concerted loss of a proton and the halide (E2, A(N)D(E)D(N)) upon base attack. Experimentally, the Taft correlation for H/D exchange, in OD(-)/D(2)O with noneliminating substrates (1-methyl-2-(2-Xethyl)pyridinium iodide), is used to predict the expected values of the rate constants for the elimination reactions with N-methylated substrates and F, Cl, Br as the leaving group. The comparison indicates an E1cb irreversible mechanism with F, but the deviation observed with Cl and Br does not allow a conclusive assignment. The theoretical calculations show that for the N methylated substrate with a fluoride leaving group the elimination proceeds via formation of a moderately stable carbanion. No stable anionic intermediate is instead found when the leaving group is Cl or Br, as well as for any of the nonmethylated species, indicating a concerted elimination. The methylated substrate with Cl shows however only a moderate increase in reactivity compared to the fluorinated substrate, despite the change in mechanism. Very interestingly, our analysis of the computed two-dimensional potential energy surface for the reaction with a F leaving group indeed evidences the lack of a net distinction between the E1cb and E2 reaction paths, which appear to merge smoothly into each other in these borderline cases. PMID- 16248657 TI - Detection of the His-heme Fe2+-NO species in the reduction of NO to N2O by ba3 oxidase from thermus thermophilus. AB - Reaction pathways in the enzymatic formation and cleavage of the N-N and N-O bonds, respectively, are difficult to verify without the structure of the intermediates, but we now have such information on the heme a(3)(2+)-NO species formed in the reaction of ba(3)-oxidase with NO from resonance Raman spectroscopy. We have identified the His-heme a(3)(2+)-NO/Cu(B)(1+) species by its characteristic Fe-NO and N-O stretching frequencies at 539 and 1620 cm(-)(1), respectively. The Fe-NO and N-O frequencies in ba(3)-oxidase are 21 and 7 cm( )(1) lower and higher, respectively, than those observed in Mb-NO. From these results and earlier Raman and FTIR measurements, we demonstrate that the protein environment of the proximal His384 that is part of the Q-proton pathway controls the strength of the Fe-His384 bond upon ligand (CO vs NO) binding. We also show by time-resolved FTIR spectroscopy that Cu(B)(1+) has a much lower affinity for NO than for CO. We suggest that the reduction of NO to N(2)O by ba(3)-oxidase proceeds by the fast binding of the first NO molecule to heme a(3) with high affinity, and the second NO molecule binds to Cu(B) with low-affinity, producing the temporal co-presence of two NO molecules in the heme-copper center. The low affinity of Cu(B) for NO binding also explains the NO reductase activity of the ba(3)-oxidase as opposed to other heme-copper oxidases. With the identification of the His-heme a(3)(2+)-NO/Cu(B)(1+) species, the structure of the binuclear heme a(3)-Cu(B)(1+) center in the initial step of the NO reduction mechanism is known. PMID- 16248659 TI - Characterization of oxygen dynamics in ZrW2O8. AB - The dynamics of oxygen motion in ZrW(2)O(8) have been characterized using (17)O solid-state NMR. Rates of dynamic exchange have been extracted from magnetization transfer experiments over a temperature range of 40 to 226 degrees C, and distinct values for the associated activation barrier have been observed on either side of the order/disorder phase transition at approximately 175 degrees C. A detailed model for the dynamical process is proposed, which reconciles the observation of continuing oxygen dynamics in the low-temperature phase with the static order implied by earlier X-ray diffraction studies. PMID- 16248658 TI - Formation and stabilization model of the 42-mer Abeta radical: implications for the long-lasting oxidative stress in Alzheimer's disease. AB - Amyloid fibrils mainly consist of 40-mer and 42-mer peptides (Abeta40, Abeta42). Abeta42 is believed to play a crucial role in the pathogenesis of Alzheimer's disease because its aggregative ability and neurotoxicity are considerably greater than those of Abeta40. The neurotoxicity of Abeta peptides involving the generation of free radicals is closely related to the S-oxidized radical cation of Met-35. However, the cation's origin and mechanism of stabilization remain unclear. Recently, structural models of fibrillar Abeta42 and Abeta40 based on systematic proline replacement have been proposed by our group [Morimoto, A.; et al. J. Biol. Chem. 2004, 279, 52781] and Wetzel's group [Williams, A. D.; et al. J. Mol. Biol. 2004, 335, 833], respectively. A major difference between these models is that our model of Abeta42 has a C-terminal beta-sheet region. Our biophysical study on Abeta42 using electron spin resonance (ESR) suggests that the S-oxidized radical cation of Met-35 could be generated by the reduction of the tyrosyl radical at Tyr-10 through a turn structure at positions 22 and 23, and stabilized by a C-terminal carboxylate anion through an intramolecular beta sheet at positions 35-37 and 40-42 to form a C-terminal core that would lead to aggregation. A time-course analysis of the generation of radicals using ESR suggests that stabilization of the radicals by aggregation might be a main reason for the long-lasting oxidative stress of Abeta42. In contrast, the S-oxidized radical cation of Abeta40 is too short-lived to induce potent neurotoxicity because no such stabilization of radicals occurs in Abeta40. PMID- 16248660 TI - Long-range electronic coupling of MM quadruple bonds (M = Mo or W) via a 2,6 azulenedicarboxylate bridge. AB - The preparation of 2,6-azulenedicarboxylic acid (I) from its diester, 2 CO(2)(t)Bu-6-CO(2)-C(10)H(6) (II), is reported together with the crystal and molecular structure of the ester, II. From the reactions between the dicarboxylic acid I and the MM quadruply bonded complexes M(2)(O(2)C(t)Bu)(4), where M = Mo or W, the azulenedicarboxylate bridged complexes [M(2)(O(2)C(t)Bu)(3)](2)(mu-2,6 (CO(2))(2)-C(10)H(6)) have been isolated, III (M = Mo) and IV (M = W). The latter compounds provide examples of electronically coupled M(2) centers via a polar bridge. The compounds show intense electronic absorptions due to metal-to-bridge charge transfer. This occurs in the visible region of the spectrum for III (M = Mo) but in the near-IR for IV (M = W). One electron oxidation with Ag(+)PF(6)(-) in THF generates the radical cations III(+) and IV(+). By both UV-vis-NIR and EPR spectroscopy the molybdenum ion III(+) is shown to be valence trapped or Class II on the Robin and Day classification scheme. Electrochemical, UV-vis-NIR, and EPR spectroscopic data indicate that, in the tungsten complex ion IV(+), the single electron is delocalized over the two W(2) centers that are separated by a distance of ca. 13.6 A. Furthermore, from the hyperfine coupling to (183)W (I = (1)/(2)), the singly occupied highest molecular orbital is seen to be polarized toward one W(2) center in relationship to the other. Electronic structure calculations employing density functional theory indicate that the HOMO in compounds III and IV is an admixture of the two M(2) delta orbitals that is largely centered on the M(2) unit having proximity to the C(5) ring of the azulenedicarboxylate bridge. The energy of the highest occupied orbital of the bridge lies very close in energy to the M(2) delta orbitals. However, this orbital does not participate in electronic coupling by a hole transfer superexchange mechanism, and the electronic coupling in the radical cations of III and IV occurs by electron transfer through the bridge pi system. PMID- 16248661 TI - Mechanism of white phosphorus activation by three-coordinate molybdenum(III) complexes: a thermochemical, kinetic, and quantum chemical investigation. AB - White phosphorus (P(4)) reacts with three-coordinate molybdenum(III) trisamides or molybdaziridine hydride complexes to produce either bridging or terminal phosphide (P(3)(-)) species, depending upon the ancillary ligand steric demands. Thermochemical measurements have been made that place the MoP triple bond dissociation enthalpy at 92.2 kcal.mol(-)(1). Thermochemical measurements together with computational analysis rule out simple P-atom abstraction from P(4) as a step in the phosphorus activation mechanism. Kinetic measurements made by the stopped-flow method show that the reaction between the monomeric molybdenum complexes and P(4) is first-order both in metal complex and in P(4). Cyclo-P(3) complexes can be obtained when ancillary ligand steric demands are small, but kinetic measurements rule them out as monometallic intermediates in the P(4) activation mechanism. Also studied by calorimetric, kinetic, and in one case variable-temperature NMR methods is the process of mu-phosphide bridge formation. Post-rate-determining steps of the P(4) activation process were examined in a search for minima on the reaction's potential energy surface, leading to the proposal of two plausible, parallel, bimetallic reaction channels. PMID- 16248662 TI - Single molecule spectroscopic investigation on conformational heterogeneity of directly linked zinc(II) porphyrin arrays. AB - We have comparatively investigated the photophysical properties of a series of meso-meso directly linked orthogonal porphyrin arrays (Zn, n = 1, 2, 3, 4, 6, 8, 9, 12, 16, 32, 48, 64, and 96) by ensemble average and single molecule fluorescence spectroscopy. In single molecule fluorescence study, we have recorded the fluorescence intensity trajectories of Zn arrays as the number of porphyrin molecules in the array increases. Up to Z8 in porphyrin arrays, each single array exhibits multiple stepwise photobleaching behaviors in fluorescence intensity trajectories, indicating that each porphyrin unit in the array acts as an individual fluorescent unit due to a maintenance of linear rigid structure of the array. On the other hand, porphyrin arrays longer than Z8 such as Z16, Z32, Z48, and Z64 show complicated photobleaching behaviors in fluorescence intensity trajectories. The origin of complex photobleaching behaviors is believed to be increasing nonradiative decay channels contributed by the enhanced structural nonlinearity in longer arrays. The fluorescence measurements of Zn arrays on single molecule level show a mismatch in the maximum fluorescence intensity level as compared to the solution measurements, which is attributable to the difference in local environment surrounding the porphyrin array. In this work, we have demonstrated the presence of conformational heterogeneity in longer porphyrin arrays by analyzing average survival times and fluorescence spectra of single arrays as the number of porphyrin molecules in the array increases. We believe that the fluorescence properties of porphyrin arrays on single molecule level will provide a platform for further applications as molecular photonic wires. PMID- 16248663 TI - Anti-thyroid drugs and thyroid hormone synthesis: effect of methimazole derivatives on peroxidase-catalyzed reactions. AB - Syntheses and characterization of the selenium analogue (MSeI) of anti-thyroid drug methimazole and a series of organoselenium compounds bearing N methylimidazole pharmacophore are described. In contrast to the sulfur compound that exists predominantly in its thione form, the selenium analogue exists in a selenol form, which spontaneously oxidizes in air to produce the corresponding diselenide. The reduction of the diselenide by GSH or NaBH(4) affords the biologically active selenol, which effectively inhibits the lactoperoxidase (LPO) activity in vitro. The monoselenides having N-methylimidazole moiety are found to be much less active than the selenol, suggesting that the presence of a selenol moiety is important for the LPO inhibition. The kinetic and mechanistic studies reveal that MSeI inhibits the LPO activity by reducing the H(2)O(2), providing a novel method to reversibly inhibit the enzyme. Although MSeI strongly inhibits LPO, the enzyme's activity can be completely recovered by increasing the H(2)O(2) concentration. On the other hand, the inhibition by methimazole (MMI), the sulfur analogue, cannot be reversed by increasing the H(2)O(2) concentration, leading to a complete inactivation of the enzyme. The reversible inhibition of LPO by some of the selenium derivatives is correlated with their glutathione peroxidase (GPx) activity, and the high GPx activity of the selenium compounds as compared with their sulfur analogues suggests that the selenium derivatives may protect the thyroid gland from oxidative damage. PMID- 16248664 TI - Dependence of field switched ordered arrays of dinuclear mixed-valence complexes on the distance between the redox centers and the size of the counterions. AB - trans-[(H(2)NCH(2)CH(2)C triple bond N)(dppe)(2)Ru(C triple bond C)(6)Ru(dppe)(2)(N triple bond CCH(2)CH(2)NH(2))][PF(6)](2), 2[PF(6)](2), a derivative of trans-[Cl(dppe)(2)Ru(C triple bond C)(6)Ru(dppe)(2)Cl] functionalized for binding to a silicon substrate, has been prepared and characterized spectroscopically, electrochemically, and with a solid state, single-crystal structure determination. Covalent binding via reaction of one amine group to a boron-doped, smooth Si-Cl substrate is verified by XPS measurements and surface electrochemistry. Vertical orientation is demonstrated by film thickness measurements. Synthesis of the 2[PF(6)](3) mixed-valence complex on the surface is established by electrochemical techniques. Measurement of the ac capacitance of the film at 1 MHz as a function of voltage across the film with a pulse-counter pulse technique demonstrates controlled electric field generation of the two stable mixed-valence forms differing in the spatial location of one electron, that is, switching. As compared to [trans-Ru(dppm)(2)(C triple bond CFc)(NCCH(2)CH(2)NH(2))][PF(6)][Cl], 1[PF(6)][Cl], the magnitude of the capacitance signal per complex observed on switching is shown to increase with increasing distance between the metal centers. Additional experiments on 1[X][Cl] show that the potential for switching 1[X][Cl] increases in the order [X](-) = [SO(3)CF(3)](-) < [PF(6)](-) < [Cl](-). A simple electrostatic model suggests that the smaller is the counterion, the greater is the perturbation of the metal sites and the larger is the barrier for switching. PMID- 16248665 TI - Magnetic field and temperature dependencies shed light on the recombination kinetics of a transition metal donor/acceptor system. AB - The radical pair recombination of an intramolecular electron-transfer system containing a transition metal moiety has been addressed by femtosecond spectroscopy. The radical pair is formed by ultrafast electron transfer (90 fs) from a ferrocene residue to a photoexcited Nile blue moiety. Its recombination proceeds on the picosecond time scale in a multiexponential fashion. The kinetic pattern is a manifestation of spin processes competing with electron transfer. Magnetic field effects on these kinetics allow one to disentangle the two contributions. Their temperature dependencies yield the activation parameters of the two processes. The discussion focuses on the mechanism of electron spin relaxation. Strong evidence for the Orbach/Kivelson mechanism will be given. PMID- 16248666 TI - Combinatorial multinuclear NMR and X-ray diffraction studies of uranium(VI) nucleotide complexes. AB - The complex formation of uranium(VI) with four nucleotides, adenosine- (AMP), guanosine- (GMP), uridine- (UMP), and cytidine-monophosphate (CMP), has been studied in the alkaline pH range (8.5-12) by (1)H, (31)P, (13)C, and (17)O NMR spectroscopy, providing spectral integral, chemical shift, homo- and heteronuclear coupling, and diffusion coefficient data. We find that two and only two complexes are formed with all ligands in the investigated pH region independently of the total uranium(VI) and ligand concentrations. Although the coordination of the 5'-phosphate group and the 2'- and 3'-hydroxyl groups of the sugar unit to the uranyl ions is similar to that proposed earlier ("Feldman complex"), the number and the structures of the complexes are different. The uranium-to-nucleotide ratio is 6:4 in one of the complexes and 3:3 in the other one, as unambiguously determined by a combinatorial approach using a systematic variation of the ratio of two ligands in ternary uranium(VI)-nucleotide systems. The structure of the 3:3 complex has been determined by single-crystal diffraction as well, and the results confirm the structure proposed by NMR in aqueous solution. The results have important implications on the synthesis of oligonucleotides. PMID- 16248668 TI - Thermoreversible cis-cisoidal to cis-transoidal isomerization of helical dendronized polyphenylacetylenes. AB - High cis content (81-99%) cis-transoidal polyphenylacetylene (PPA) jacketed with amphiphilic self-assembling dendrons, poly[(3,4-3,5)mG2-4EBn] with m = 8, 10, 12, 14, 16, and (S)-3,7-dimethyloctyl, were synthesized by Rh(C triple bond CPh)(nbd)(PPh(3))(2) (nbd = 2,5-norbornadiene)/N,N-(dimethylamino)pyridine (DMAP) catalyzed polymerization of macromonomers. The resulting cylindrical PPAs self organize into hexagonal columnar lattices with intracolumnar order (Phi(h)(io)) and without (Phi(h)). The polymers with m = 12, 14, and 16 exhibit also a hexagonal columnar crystal phase (Phi(h,k)). The reversible Phi(h,k)-to Phi(h)(io)-to- Phi(h) phase transition in these dendronized PPAs was analyzed by a combination of differential scanning calorimetry and small and wide-angle X-ray diffraction experiments performed on powder and oriented fibers. In the Phi(h,k) and Phi(h)(io) phases, the dendronized PPAs form helical porous columns. The helical pore disappears in the Phi(h) phase. This change is accompanied by a decrease of the external column diameter that is induced by stretching of the polymer backbone along the axis of the cylinder. The helix sense of the porous PPA is selected by homochiral alkyl dendritic tails. This transition is generated by an unprecedented conversion of the PPA backbone from the cis-cisoidal conformation in the Phi(h,k) and Phi(h)(io) phases to the cis-transoidal conformation in the Phi(h) phase. Under the same conditions, the pristine cis-PPA undergoes cis-trans isomerization and irreversible intramolecular 6pi electrocyclization of 1,3-cis,5-hexatriene sequences followed by chain cleavage. These processes are eliminated in the dendronized cis-PPA below its decomposition temperature. PMID- 16248667 TI - Role of protein dynamics in reaction rate enhancement by enzymes. AB - An integrated view of protein structure, dynamics, and function is emerging, where proteins are considered as dynamically active assemblies and internal motions are closely linked to function such as enzyme catalysis. Further, the motion of solvent bound to external regions of protein impacts internal motions and, therefore, protein function. Recently, we discovered a network of protein vibrations in enzyme cyclophilin A, coupled to its catalytic activity of peptidyl prolyl cis-trans isomerization. Detailed studies suggest that this network, extending from surface regions to active site, is a conserved part of enzyme structure and has a role in promoting catalysis. In this report, theoretical investigations of concerted conformational fluctuations occurring on microsecond and longer time scales within the discovered network are presented. Using a new technique, kinetic energy was added to protein vibrational modes corresponding to conformational fluctuations in the network. The results reveal that protein dynamics promotes catalysis by altering transition state barrier crossing behavior of reaction trajectories. An increase in transmission coefficient and number of productive trajectories with increasing amounts of kinetic energy in vibrational modes is observed. Variations in active site enzyme-substrate interactions near transition state are found to be correlated with barrier recrossings. Simulations also showed that energy transferred from first solvation shell to surface residues impacts catalysis through network fluctuations. The detailed characterization of network presented here indicates that protein dynamics plays a role in rate enhancement by enzymes. Therefore, coupled networks in enzymes have wide implications in understanding allostericity and cooperative effects, as well as protein engineering and drug design. PMID- 16248669 TI - A general method for preparation of metal carbenes via solution- and polymer based approaches. AB - A new general, synthetically simple, and safe method for the preparation of metal carbene complexes, which is based on diphenyl sulfonium salts as carbenoid precursors, has been developed, and its scope and applications were studied. In general, deprotonation of a sulfonium salt with a base results in a sulfur ylide, which, in turn, reacts with an appropriate metal precursor to give the corresponding metal carbene complex. Thus, starting from benzyldiphenylsulfonium salt, the complexes (PCX)Rh=CHPh (X = P, N) were prepared in quantitative yield. Syntheses of Grubbs' catalyst, (PCy(3))(2)Cl(2)Ru=CHPh, and of Werner's carbene, [Os(=CHPh)HCl(CO)(P(i)Pr(3))(2)], were achieved by this method. Novel trans bisphosphine Rh and Ir carbenes, ((i)Pr(3)P)(2)(Cl)M=CHPh, which could not be prepared by other known methods, were synthesized by the sulfur ylide approach. The method is not limited to metal benzylidenes, as demonstrated by the preparation of the Ru vinyl-alkylidene, (PCy(3))(2)Cl(2)Ru=CH-CH=CH(2), methoxycarbonyl-alkylidene, (PCy(3))(2)Cl(2)Ru=CH(CO(2)Me), and alkylidene (PCy(3))(2)Cl(2)Ru=CH(CH(3)), (PCy(3))(2)Cl(2)Ru=CH(2) compounds. The problem of recycling of starting materials as well as the issue of facile purification of the product metal carbene complex were addressed by the synthesis of a polymer supported diarylsulfide, the carrier of the carbenoid unit in the process. Based on the sulfur ylide route, a methodology for the synthesis of metallocarbenes anchored to a polymer via the carbene ligand, using a commercial Merrifield resin, was developed. PMID- 16248671 TI - Electronic relaxation dynamics of water cluster anions. AB - The electronic relaxation dynamics of water cluster anions, (H(2)O)(n)(-), have been studied with time-resolved photoelectron imaging. In this investigation, the excess electron was excited through the p<--s transition with an ultrafast laser pulse, with subsequent electronic evolution monitored by photodetachment. All excited-state lifetimes exhibit a significant isotope effect (tau(D)2(O)/tau(H)2(O) approximately 2). Additionally, marked dynamical differences are found for two classes of water cluster anions, isomers I and II, previously assigned as clusters with internally solvated and surface-bound electrons, respectively. Isomer I clusters with n > or = 25 decay exclusively by internal conversion, with relaxation times that extrapolate linearly with 1/n toward an internal conversion lifetime of 50 fs in bulk water. Smaller isomer I clusters (13 < or = n < or = 25) decay through a combination of excited-state autodetachment and internal conversion. The relaxation of isomer II clusters shows no significant size dependence over the range of n = 60-100, with autodetachment an important decay channel following excitation of these clusters. Photoelectron angular distributions (PADs) were measured for isomer I and isomer II clusters. The large differences in dynamical trends, relaxation mechanisms, and PADs between large isomer I and isomer II clusters are consistent with their assignment to very different electron binding motifs. PMID- 16248670 TI - The structure of a mixed LNA/DNA:RNA duplex is driven by conformational coupling between LNA and deoxyribose residues as determined from 13C relaxation measurements. AB - A study of the internal dynamics of an LNA/DNA:RNA duplex has been performed to further characterize the conformational changes associated with the incorporation of locked nucleic acid (LNA) nucleotides in a DNA:RNA duplex. In general, it was demonstrated that the LNA/DNA:RNA duplex has a very high degree of order compared to dsDNA and dsRNA duplexes. The order parameters of the aromatic carbon atoms in the LNA/DNA strand are uniformly high, whereas a sharp drop in the degree of order was seen in the RNA strand in the beginning of the AUAU stretch in the middle of the strand. This can be related to a return to normal dsRNA dynamics for the central A:U base pair. The high order of the heteroduplex is consistent with preorganization of the chimera strand for an A-form duplex conformation. These results partly explain the dramatic increase in T(m) of the chimeric heteroduplex over dsDNA and DNA:RNA hybrids of the same sequence. PMID- 16248673 TI - The F/Ph rearrangement reaction of [(Ph3P)3RhF], the fluoride congener of Wilkinson's catalyst. AB - The fluoride congener of Wilkinson's catalyst, [(Ph(3)P)(3)RhF] (1), has been synthesized and fully characterized. Unlike Wilkinson's catalyst, 1 easily activates the inert C-Cl bond of ArCl (Ar = Ph, p-tolyl) under mild conditions (3 h at 80 degrees C) to produce trans-[(Ph(3)P)(2)Rh(Ph(2)PF)(Cl)] (2) and ArPh as a result of C-Cl, Rh-F, and P-C bond cleavage and C-C, Rh-Cl, and P-F bond formation. In benzene (2-3 h at 80 degrees C), 1 decomposes to a 1:1 mixture of trans-[(Ph(3)P)(2)Rh(Ph(2)PF)(F)] (3) and the cyclometalated complex [(Ph(3)P)(2)Rh(Ph(2)PC(6)H(4))] (4). Both the chloroarene activation and the thermal decomposition reactions have been shown to occur via the facile and reversible F/Ph rearrangement reaction of 1 to cis-[(Ph(3)P)(2)Rh(Ph)(Ph(2)PF)] (5), which has been isolated and fully characterized. Kinetic studies of the F/Ph rearrangement, an intramolecular process not influenced by extra phosphine, have led to the determination of E(a) = 22.7 +/- 1.2 kcal mol(-)(1), DeltaH(++) = 22.0 +/- 1.2 kcal mol(-)(1), and DeltaS(++) = -10.0 +/- 3.7 eu. Theoretical studies of F/Ph exchange with the [(PH(3))(2)(PH(2)Ph)RhF] model system pointed to two possible mechanisms: (i) Ph transfer to Rh followed by F transfer to P (formally oxidative addition followed by reductive elimination, pathway 1) and (ii) F transfer to produce a metallophosphorane with subsequent Ph transfer to Rh (pathway 2). Although pathway 1 cannot be ruled out completely, the metallophosphorane mechanism finds more support from both our own and previously reported observations. Possible involvement of metallophosphorane intermediates in various P-F, P-O, and P-C bond-forming reactions at a metal center is discussed. PMID- 16248672 TI - Michael addition reactions between chiral equivalents of a nucleophilic glycine and (S)- or (R)-3-[(E)-enoyl]-4-phenyl-1,3-oxazolidin-2-ones as a general method for efficient preparation of beta-substituted pyroglutamic acids. Case of topographically controlled stereoselectivity. AB - This paper describes a systematic study of addition reactions between the chiral Ni(II) complex of the Schiff base of glycine with (S)-o-[N-(N benzylprolyl)amino]benzophenone and (S)- or (R)-3-[(E)-enoyl]-4-phenyl-1,3 oxazolidin-2-ones as a general and synthetically efficient approach to beta substituted pyroglutamic acids and relevant compounds. These reactions were shown to occur at room temperature in the presence of nonchelating organic bases and, most notably, with very high (>98% diastereomeric excess (de)) stereoselectivity at both newly formed stereogenic centers. The stereochemical outcome of the reactions was found to be overwhelmingly controlled by the stereochemical preferences of the Michael acceptors, and the chirality of the glycine complex influenced only the reaction rate. Thus, in the reactions of both the (S) configured Ni(II) complex and the Michael acceptors, the reaction rates were exceptionally high, allowing preparation of the corresponding products with virtually quantitative (>98%) chemical and stereochemical yields. In contrast, reactions of the (S)-configured Ni(II) complex and (R)-configured Michael acceptors proceeded at noticeably lower rates, but the addition products were obtained in high diastereo- and enantiomeric purity. To rationalize the remarkably high and robust stereoselectivity observed in these reactions, we consider an enzyme-substrate-like mode of interaction involving a topographical match or mismatch of two geometric figures. Excellent chemical and stereochemical yields, combined with the simplicity and operational convenience of the experimental procedures, render the present method of immediate use for preparing various beta-substituted pyroglutamic acids and related compounds. PMID- 16248674 TI - Bismuth spheres grown in self-nested cavities in a silicon wafer. AB - We have developed a one-step, hydrofluoric acid-free hydrothermal etching method that not only produces bismuth nano/micrometer-sized spheres but also prepares porous silicon with vertical holes. By controlling the heating temperature and time, nanoscale vertical-channeled porous silicon can be received. Our result indicated that the Bi clusters were formed first on the wafer surface. Then the etching of the Bi to the wafer creates the holes. Later, the Bi spheres went into the holes and expedited the etching process. A formation mechanism and chemical process have been proposed on the basis of experimental data. This simple chemistry approach may be of great scientific and technological importance for preparing porous silicon wafer. PMID- 16248676 TI - Exploitative and hierarchical antagonism in a cooperative bacterium. AB - Social organisms that cooperate with some members of their own species, such as close relatives, may fail to cooperate with other genotypes of the same species. Such noncooperation may take the form of outright antagonism or social exploitation. Myxococcus xanthus is a highly social prokaryote that cooperatively develops into spore-bearing, multicellular fruiting bodies in response to starvation. Here we have characterized the nature of social interactions among nine developmentally proficient strains of M. xanthus isolated from spatially distant locations. Strains were competed against one another in all possible pairwise combinations during starvation-induced development. In most pairings, at least one competitor exhibited strong antagonism toward its partner and a majority of mixes showed bidirectional antagonism that decreased total spore production, even to the point of driving whole populations to extinction. Differential response to mixing was the primary determinant of competitive superiority rather than the sporulation efficiencies of unmixed populations. In some competitive pairings, the dominant partner sporulated more efficiently in mixed populations than in clonal isolation. This finding represents a novel form of exploitation in bacteria carried out by socially competent genotypes and is the first documentation of social exploitation among natural bacterial isolates. Patterns of antagonistic superiority among these strains form a highly linear dominance hierarchy. At least some competition pairs construct chimeric, rather than segregated, fruiting bodies. The cooperative prokaryote M. xanthus has diverged into a large number of distinct social types that cooperate with clone mates but exhibit intense antagonism toward distinct social types of the same species. Most lengthy migration events in nature may thus result in strong antagonism between migratory and resident populations, and this antagonism may have large effects on local population sizes and dynamics. Intense mutual antagonism appears to be more prevalent in this prokaryotic social species than has been observed in the eukaryotic social slime mold Dictyostelium discoideum, which also exhibits multicellular development. The finding of several cases of facultative social exploitation among these natural isolates suggests that such exploitation may occur frequently in nature in many prokaryotes with cooperative traits. PMID- 16248677 TI - The case for selection at CCR5-Delta32. AB - The C-C chemokine receptor 5, 32 base-pair deletion (CCR5-Delta32) allele confers strong resistance to infection by the AIDS virus HIV. Previous studies have suggested that CCR5-Delta32 arose within the past 1,000 y and rose to its present high frequency (5%-14%) in Europe as a result of strong positive selection, perhaps by such selective agents as the bubonic plague or smallpox during the Middle Ages. This hypothesis was based on several lines of evidence, including the absence of the allele outside of Europe and long-range linkage disequilibrium at the locus. We reevaluated this evidence with the benefit of much denser genetic maps and extensive control data. We find that the pattern of genetic variation at CCR5-Delta32 does not stand out as exceptional relative to other loci across the genome. Moreover using newer genetic maps, we estimated that the CCR5-Delta32 allele is likely to have arisen more than 5,000 y ago. While such results can not rule out the possibility that some selection may have occurred at C-C chemokine receptor 5 (CCR5), they imply that the pattern of genetic variation seen at CCR5-Delta32 is consistent with neutral evolution. More broadly, the results have general implications for the design of future studies to detect the signs of positive selection in the human genome. PMID- 16248678 TI - Evolution of mutational robustness in an RNA virus. AB - Mutational (genetic) robustness is phenotypic constancy in the face of mutational changes to the genome. Robustness is critical to the understanding of evolution because phenotypically expressed genetic variation is the fuel of natural selection. Nonetheless, the evidence for adaptive evolution of mutational robustness in biological populations is controversial. Robustness should be selectively favored when mutation rates are high, a common feature of RNA viruses. However, selection for robustness may be relaxed under virus co infection because complementation between virus genotypes can buffer mutational effects. We therefore hypothesized that selection for genetic robustness in viruses will be weakened with increasing frequency of co-infection. To test this idea, we used populations of RNA phage phi6 that were experimentally evolved at low and high levels of co-infection and subjected lineages of these viruses to mutation accumulation through population bottlenecking. The data demonstrate that viruses evolved under high co-infection show relatively greater mean magnitude and variance in the fitness changes generated by addition of random mutations, confirming our hypothesis that they experience weakened selection for robustness. Our study further suggests that co-infection of host cells may be advantageous to RNA viruses only in the short term. In addition, we observed higher mutation frequencies in the more robust viruses, indicating that evolution of robustness might foster less-accurate genome replication in RNA viruses. PMID- 16248679 TI - Variant histone H2A.Z is globally localized to the promoters of inactive yeast genes and regulates nucleosome positioning. AB - H2A.Z is an evolutionary conserved histone variant involved in transcriptional regulation, antisilencing, silencing, and genome stability. The mechanism(s) by which H2A.Z regulates these various biological functions remains poorly defined, in part due to the lack of knowledge regarding its physical location along chromosomes and the bearing it has in regulating chromatin structure. Here we mapped H2A.Z across the yeast genome at an approximately 300-bp resolution, using chromatin immunoprecipitation combined with tiling microarrays. We have identified 4,862 small regions--typically one or two nucleosomes wide--decorated with H2A.Z. Those "Z loci" are predominantly found within specific nucleosomes in the promoter of inactive genes all across the genome. Furthermore, we have shown that H2A.Z can regulate nucleosome positioning at the GAL1 promoter. Within HZAD domains, the regions where H2A.Z shows an antisilencing function, H2A.Z is localized in a wider pattern, suggesting that the variant histone regulates a silencing and transcriptional activation via different mechanisms. Our data suggest that the incorporation of H2A.Z into specific promoter-bound nucleosomes configures chromatin structure to poise genes for transcriptional activation. The relevance of these findings to higher eukaryotes is discussed. PMID- 16248680 TI - Ultrasonic songs of male mice. AB - Previously it was shown that male mice, when they encounter female mice or their pheromones, emit ultrasonic vocalizations with frequencies ranging over 30-110 kHz. Here, we show that these vocalizations have the characteristics of song, consisting of several different syllable types, whose temporal sequencing includes the utterance of repeated phrases. Individual males produce songs with characteristic syllabic and temporal structure. This study provides a quantitative initial description of male mouse songs, and opens the possibility of studying song production and perception in an established genetic model organism. PMID- 16248682 TI - Chronicity of psychological strain in occupational settings and the accuracy of the General Health Questionnaire. AB - This article examines the proposition that the traditional scoring method of the General Health Questionnaire (GHQ) underestimates the number of respondents classified as "cases." A revised "chronic" scoring method (the CGHQ) is used and demonstrates superior construct validity and greater sensitivity. A comparison of the CGHQ and GHQ also shows the CGHQ to be a superior criterion measure. These claims are demonstrated by survey data gathered from 3 occupational samples (Ns=11,637, 2,253, and 2,124). Results show that the CGHQ is more appropriate as a screening instrument for psychological morbidity. Tests of construct validity also favor the CGHQ with only a slight advantage for predictive validity in terms of variance explained. The more desirable statistical properties of the CGHQ result in a reduction of significant interaction terms and are strongly recommended in future studies as a means of controlling Type I errors when tests of moderation are examined. PMID- 16248683 TI - Countering the negative effects of job insecurity through participative decision making: lessons from the demand-control model. AB - This study examined the effectiveness of increased organizational participative decision making in attenuating the negative consequences of job insecurity. Data were collected from 807 employees in 6 different companies. Analyses suggest that job insecurity is related to lower coworker, work, and supervisor satisfaction and higher turnover intentions and work withdrawal behaviors. However, employees with greater participative decision-making opportunities reported fewer negative consequences of job insecurity compared with employees with fewer participative decision-making opportunities. Results are interpreted using the demand-control model and suggest that organizations that allow greater employee participative decision making may experience fewer negative side effects from today's rising levels of employee job insecurity. PMID- 16248685 TI - The association between burnout, depression, anxiety, and inflammation biomarkers: C-reactive protein and fibrinogen in men and women. AB - Following the demonstrated association of employee burnout or vital exhaustion with several risk factors for cardiovascular disease (CVD) and CVD risk, the authors investigated the possibility that one of the mechanisms linking burnout with CVD morbidity is microinflammation, gauged in this study by high-sensitivity C-reactive protein (hs-CRP) and fibrinogen concentrations. Their sample included 630 women and 933 men, all apparently healthy, who underwent periodic health examinations. The authors controlled for possible confounders including 2 other negative affective states: depression and anxiety. In women, burnout was positively associated with hs-CRP and fibrinogen concentrations, and anxiety was negatively associated with them. In men, depression was positively associated with hs-CRP and fibrinogen concentrations, but not with burnout or anxiety. Thus, burnout, depression, and anxiety are differentially associated with microinflammation biomarkers, dependent on gender. PMID- 16248684 TI - The relationship between perceptions of politics and depressed mood at work: unique moderators across three levels. AB - The authors examined the unique moderating potential of need for achievement, perceived organizational support, and faith in management on the relationship between perceptions of politics across 3 hierarchical levels (one's peer level, 1 level up, and at the highest level in the organization) and depressed mood at work. Results from 173 full-time employees, representing a wide range of occupations, supported the hypotheses. Specifically, the authors found that need for achievement interacted with perceived politics at one's peer level, perceived organizational support interacted with perceived politics at 1 level up, and faith in management interacted with politics perceived at the highest levels in the organization to relate to depressed mood at work. Contributions of this study, strengths and limitations, and future research directions are provided. PMID- 16248686 TI - The Physical Health Questionnaire (PHQ): construct validation of a self-report scale of somatic symptoms. AB - The authors report the results of 3 studies that were conducted to evaluate the psychometric properties of the Physical Health Questionnaire (PHQ), a brief self report scale of somatic symptoms. In Study 1, exploratory factor analysis results revealed 4 empirically distinct dimensions of somatic symptoms: gastrointestinal problems, headaches, sleep disturbances, and respiratory illness. In Study 2, this structure was replicated using confirmatory factor analysis, and correlations of the PHQ dimensions with measures of negative affect, psychological health, and job performance provided further validity evidence. In Study 3, a minor revision to the wording of several items helped to address the limitations of one of the PHQ subscales. Together, these results provide evidence of the construct validity of the PHQ. PMID- 16248687 TI - Children's health and workers' productivity: an examination of family interference with work in rural America. AB - This article examines the association of children's health with their parents' performance in the workplace using data from a random survey sample of adults living in rural western North Carolina (N=206). Guided by the effort-recovery model, the authors hypothesized that parents whose children are more ill have poorer performance in the workplace because their parenting requires greater effort and they have less opportunity for physical and psychological recovery. Child health was unassociated with parents cutting back at work because of physical health. Poorer child health was associated with parents cutting back at work because of emotional health, and a portion of this association, as hypothesized, was explained by more limited opportunities for parental recovery. There was no evidence suggesting that associations differed by parental gender. PMID- 16248689 TI - Daily affect and daily beliefs. AB - Human resource directorate employees of a large United Kingdom public hospital (N=36) completed an initial questionnaire and then participated in a daily diary study. The questionnaire included measures of affect and beliefs about high work demands' influence on affect and work performance. The diary included measures of affect, extent of high work demands, and daily beliefs, corresponding to those measured in the questionnaire. Participants were required to complete the diary twice daily, before and after work over a 2-week period. Measures of affect after work were associated with beliefs concerning work demands' influence on work performance and on affect measured after work. Beliefs about work demands measured in the questionnaire were associated with subsequent daily assessments of beliefs. PMID- 16248688 TI - Switching off mentally: predictors and consequences of psychological detachment from work during off-job time. AB - Psychological detachment from work refers to the off-job experience of "switching off" mentally. It is hypothesized that a high degree of workload encountered during the work day has a negative impact on subsequent detachment processes and that psychological detachment from work is positively related to well-being. Eighty-seven individuals from various occupations provided questionnaire and daily survey measures over a period of 3 working days. Multilevel analysis showed that workload was negatively related to psychological detachment from work during evening hours. Psychological detachment from work was associated with positive mood and low fatigue. The negative relationship between psychological detachment and fatigue was particularly strong on days with high time pressure. PMID- 16248690 TI - Health-related quality of life measured by the SF12 in working populations: associations with psychosocial work characteristics. AB - This study investigated the contribution of psychosocial work characteristics (decision latitude, job demand, social support at work, and effort-reward imbalance) to health-related quality of life. Data were derived from 2 aircraft manufacturing plants (N=1,855) at the start of a longitudinal study. Regression analysis showed that work characteristics (1st model) explained 19% of the variance in the mental summary score of the Short Form-12 Health Survey. R2 change for work characteristics decreased to 13%, accounting for demographics, socioeconomic status, body mass index, and medical condition (5th model). Including health behavior and personality factors (full model), R2 change for work characteristics remained significant. Psychosocial work characteristics account for relevant proportions in the subjective perception of mental health beyond a wide array of medical variables and personality factors. PMID- 16248691 TI - Core self-evaluations and job burnout: the test of alternative models. AB - Research on job burnout has traditionally focused on contextual antecedent conditions, although a theoretically appropriate conception implicates person environment relationships. The authors tested several models featuring various combinations of personal and contextual influences on burnout and job satisfaction. Measures of core self-evaluations, organizational constraints, burnout, and job satisfaction were collected from 859 health care employees. Results from structural equations modeling analyses revealed an influence of core self-evaluations and perceived organizational constraints on job burnout and satisfaction, suggesting personal and contextual contributions. These results favor a broadening of current thinking about the impact of situational constraints on the expression of job burnout, as well as for the role of disposition for affective responding to effectively address occupational health problems. PMID- 16248692 TI - Extending and applying the demand-control model: the role of soldier's coping on a peacekeeping deployment. AB - The purpose of this study was to extend the demand-control model (R. A. Karasek, 1979) by examining coping as an additional factor. It was hypothesized that perceived job control only buffered the demand-strain relationship when individuals used active coping and exacerbated the relationship when individuals used passive coping. Soldiers (N=638) were surveyed before and during a 6-month peacekeeping deployment to Kosovo. Results partially confirmed the hypotheses. Even after controlling for general psychological health at predeployment, job control moderated the relationship between demands and psychological health during deployment when soldiers used active coping. No significant 3-way interactions were found for religious coping and passive coping. Implications for demand-control modeling and potential applications of the findings to soldier and leader training are discussed. PMID- 16248693 TI - Moderators of the relationship between long work hours and health. AB - The authors examined how associations between work hours and self-reported health are moderated by the reasons given for working overtime, by schedule autonomy, and by the degree of social support experienced, in a survey of 372 train drivers. Respondents who lacked both schedule autonomy and social support demonstrated positive associations between the number of hours worked per week and frequency of physical health symptoms. Conversely, negative associations were observed among respondents reporting low schedule autonomy together with high social support. There were no such interactions in the analyses of fatigue and psychological health outcomes. The findings are discussed in terms of the fit between an individual's actual and desired work hours. The problems of identifying appropriate limits for work hours are highlighted. PMID- 16248694 TI - Development and validation of the uncivil workplace behavior questionnaire. AB - This article describes the development and validation of a new measure of workplace incivility, the Uncivil Workplace Behavior Questionnaire (UWBQ). Participants included 5 samples of Australian adult employees (total N=368). Principal axis factoring of the UWBQ yielded 4 interpretable factors (Hostility, Privacy Invasion, Exclusionary Behavior, and Gossiping), all of which exhibited high internal consistency. The 4-factor structure received further support from a confirmatory factor analysis on a hold-out sample. A series of correlation and regression analyses revealed that the UWBQ subscales exhibited sound convergent, divergent, and concurrent validity. The psychometric properties of the UWBQ are contrasted with those of the Workplace Incivility Scale (L. M. Cortina, V. J. Magley, J. H. Williams, & R. D. Langhout, 2001), to the authors' knowledge the only other measure of the workplace incivility construct available to date. PMID- 16248695 TI - Consequences of boundary-spanning demands and resources for work-to-family conflict and perceived stress. AB - Using work-family border theory, this article examines relationships between boundary-spanning demands and resources and work-to-family conflict and perceived stress. The analysis uses data from 2,109 respondents from the 2002 National Study of the Changing Workforce. The demands that were positively related to work to-family conflict and perceived stress were commuting time, bringing work home, job contacts at home, and work-family multitasking. Work-family multitasking partially explained the effects of bringing work home and job contacts at home on conflict and stress. For resources, time off for family responsibilities and a supportive work-family culture showed negative associations with conflict and stress. Work-to-family conflict partially mediated relationships between several demands and resources and perceived stress. PMID- 16248696 TI - Railing for safety: job demands, job control, and safety citizenship role definition. AB - This study investigated job demands and job control as predictors of safety citizenship role definition, that is, employees' role orientation toward improving workplace safety. Data from a survey of 334 trackside workers were framed in the context of R. A. Karasek's (1979) job demands-control model. High job demands were negatively related to safety citizenship role definition, whereas high job control was positively related to this construct. Safety citizenship role definition of employees with high job control was buffered from the influence of high job demands, unlike that of employees with low job control, for whom high job demands were related to lower levels of the construct. Employees facing both high job demands and low job control were less likely than other employees to view improving safety as part of their role orientation. PMID- 16248697 TI - The implications of cortical recruitment and brain morphology for individual differences in inhibitory function in aging humans. AB - The authors assessed individual differences in cortical recruitment, brain morphology, and inhibitory task performance. Similar to previous studies, older adults tended toward bilateral activity during task performance more than younger adults. However, better performing older adults showed less bilateral activity than poorer performers, contrary to the idea that additional activity is universally compensatory. A review of the results and of extant literature suggests that compensatory activity in prefrontal cortex may only be effective if the additional cortical processors brought to bear on the task can play a complementary role in task performance. Morphological analyses revealed that frontal white matter tracts differed as a function of performance in older adults, suggesting that hemispheric connectivity might impact both patterns of recruitment and cognitive performance. PMID- 16248698 TI - Age and regulatory focus determine preferences for health-related role models. AB - The authors hypothesized that the effectiveness of role models varies across the adult life span because of differences in health-related regulatory orientations. Because young adults have strong health-related promotion orientations, they should be motivated by positive models who illustrate the benefits of good health. Because older adults have more balanced health-related promotion and prevention orientations, they should be motivated not only by positive models but also by negative models who illustrate the costs of poor health. Results indicated that both young and older adults perceived positive models to be motivating, but older adults found negative models to be more motivating than did young adults. Age differences in responses to negative models were partially mediated by differences in health-related prevention orientation. PMID- 16248699 TI - Intraindividual variability may not always indicate vulnerability in elders' cognitive performance. AB - This study examined consistency of performance, or intraindividual variability, in older adults' performance on 3 measures of cognitive functioning: inductive reasoning, memory, and perceptual speed. Theoretical speculation has suggested that such intraindividual variability may signal underlying vulnerability or neurologic compromise. Thirty-six participants aged 60 and older completed self administered cognitive assessments twice a day for 60 consecutive days. Intraindividual variability was not strongly correlated among the 3 cognitive measures, but, over the course of the study, intraindividual variability was strongly intercorrelated within a task. Higher average performance on a measure was associated with greater performance variability, and follow-up analyses revealed that a higher level of intraindividual variability is positively associated with the magnitude of a person's practice-related gain on a particular measure. The authors argue that both adaptive (practice-related) and maladaptive (inconsistency-related) intraindividual variability may exist within the same individuals over time. PMID- 16248700 TI - Psychophysiological mediators of caregiver stress and differential cognitive decline. AB - The authors examined relationships between chronic stress and cognitive decline and whether such relationships were mediated by psychophysiological factors. Ninety-six caregivers of spouses with Alzheimer's disease (AD) were compared with 95 similar noncaregiver spouses. All were free of diabetes. Although the groups started similarly, over 2 years caregivers declined by a small but significant amount (1 raw score point and 4 percentile points, each p<.05) on Shipley Vocabulary. In contrast, noncaregivers did not change. Higher hostile attribution (beta=-.09; p<.05) and metabolic risk (beta=-.10; p<.05) in caregivers mediated the cognitive decline. This is the first study of cognitive decline and mediators in caregivers. This work has implications for caregiver and care-recipient health and for research on cognition, psychophysiology, diabetes, and AD. PMID- 16248701 TI - Multivariate modeling of age and retest in longitudinal studies of cognitive abilities. AB - Longitudinal multivariate mixed models were used to examine the correlates of change between memory and processing speed and the contribution of age and retest to such change correlates. Various age- and occasion-mixed models were fitted to 2 longitudinal data sets of adult individuals (N>1,200). For both data sets, the results indicated that the correlation between the age slopes of memory and processing speed decreased when retest effects were included in the model. If retest effects existed in the data but were not modeled, the correlation between the age slopes was positively biased. The authors suggest that although the changes in memory and processing speed may be correlated over time, age alone does not capture such a covariation. PMID- 16248702 TI - Social participation attenuates decline in perceptual speed in old and very old age. AB - Does an engaged and active lifestyle in old age alleviate cognitive decline, does high cognitive functioning in old age increase the possibility of maintaining an engaged and active lifestyle, or both? The authors approach this conundrum by applying a structural equation model for testing dynamic hypotheses, the dual change score model (J. J. McArdle & F. Hamagami, 2001), to 3-occasion longitudinal data from the Berlin Aging Study (Time 1: n=516, age range=70-103 years). Results reveal that within a bivariate system of perceptual speed and social participation, with age and sociobiographical status as covariates, prior scores of social participation influence subsequent changes in perceptual speed, while the opposite does not hold. Results support the hypothesis that an engaged and active lifestyle in old and very old age may alleviate decline in perceptual speed. PMID- 16248704 TI - Age and experience influences on the complexity of social inferences. AB - Age differences in social-cognitive functioning were assessed by examining sensitivity to the trait implications of behavioral cues when making social inferences. Adults (age range = 23-86 years) read target descriptions containing positive and negative behaviors relating to either morality or competence. Consistent with past research, middle-aged and older adults were more likely than younger adults to make inferences consistent with the trait-diagnostic implications of the behaviors. Age was also associated with increased sensitivity to additional cues that moderated the diagnostic value of behaviors based on simple descriptive content. The authors argue that these age differences reflect a type of expertise based in accumulated social experience, a conclusion bolstered by an additional finding that social activity moderated age differences in social judgments. PMID- 16248703 TI - Age-related differences in the processing of redundant visual dimensions. AB - Age differences in the redundant-signals effect and coactivation of visual dimensions were investigated in 2 experiments. In Experiment 1 the task required the conjoining of dimensions, whereas in Experiment 2 the spatial separation of dimensions was manipulated. Although coactivation was evident for both age groups when the redundant dimensions occurred at the same location, older adults showed more evidence for coactivation, perhaps because of compensation for declines in perceptual processing. When the redundant dimensions were separated, neither age group showed evidence for coactivation. These findings indicate that the coactive processing of redundant visual dimensions is spared in healthy older adults and that for both groups, attention must be focused on both dimensions for coactivation to occur. PMID- 16248705 TI - Estimating age change in list recall in asset and health dynamics of the oldest old: the effects of attrition bias and missing data treatment. AB - Average change in list recall was evaluated as a function of missing data treatment (Study 1) and dropout status (Study 2) over ages 70 to 105 in Asset and Health Dynamics of the Oldest-Old data. In Study 1 the authors compared results of full-information maximum likelihood (FIML) and the multiple imputation (MI) missing-data treatments with and without independent predictors of missingness. Results showed declines in all treatments, but declines were larger for FIML and MI treatments when predictors were included in the treatment of missing data, indicating that attrition bias was reduced. In Study 2, models that included dropout status had better fits and reduced random variance compared with models without dropout status. The authors conclude that change estimates are most accurate when independent predictors of missingness are included in the treatment of missing data with either MI or FIML and when dropout effects are modeled. PMID- 16248706 TI - Cardiovascular intraindividual variability in later life: the influence of social connectedness and positive emotions. AB - Healthy normotensive men and women (N=33) underwent a 60-day diary assessment of emotions and cardiovascular functioning. Individual differences in social connectedness and mood were measured in questionnaires, and positive emotions, negative emotions, systolic blood pressure (SBP), and diastolic blood pressure (DBP) were assessed daily for 60 consecutive days. Results confirmed that the cardiovascular undoing effect of positive emotions is evident primarily in the context of negative emotional arousal. The daily associations between positive emotions and cardiovascular outcomes were linked to individual differences in social connectedness. Controlling for individual differences in mood levels, multilevel regression analyses showed that social connectedness predicted extended positive emotion, diminished SBP and DBP reactivity, and more rapid SBP recovery from daily negative emotional states. PMID- 16248707 TI - Adult age differences in distinctive processing: the modality effect on false recall. AB - Age differences in distinctive processing were investigated by examining the effects of study presentation modality on false recall in younger and older adults using the Deese/Roediger and McDermott paradigm. Participants were presented with study words either visually or auditorily. Older adults did not show the typical reduction in false recall after visual, compared to auditory, study presentation (R.E. Smith & R.R. Hunt, 1998). The authors interpret these results as evidence of reduced distinctive processing on the part of older adults. PMID- 16248709 TI - Symptoms of depression in older home-care patients: patient and informant reports. AB - The purpose of this study was to examine the level of agreement and patterns of disagreement between home-care patient and informant reports of depressive symptoms. The authors interviewed a sample of 355 older home-care patients and their informants using the Structured Diagnostic Interview for DSM-IV (R. L. Spitzer, M. Gibbon, & J. B. Williams, 1995). Informants reported more psychological symptoms than patients, and this type of discrepancy was higher for patients with cognitive impairment and patients who had younger informants. Younger informants also reported more cognitive symptoms, whereas patients were more likely to report suicidal thoughts or ideation if they were not cognitively impaired. The patterns of these discrepancies may reflect age- and cohort-related response bias in the reports of depressive symptoms obtained from older adults. PMID- 16248710 TI - Aging, inhibition, and social inappropriateness. AB - This study explores the hypothesis that age-related declines in inhibitory ability are associated with increases in socially inappropriate behavior. Consistent with this hypothesis, older participants were less likely than younger participants to differentiate between public and private settings when inquiring about potentially embarrassing issues, according to their peers. Additionally, this indiscriminate public inquiry was associated with decreased closeness with participants' peers, particularly for older adults. Finally, this age-related increase in social inappropriateness was mediated by inhibitory deficits associated with aging. These results suggest that age-related deficits in inhibitory ability may cause people to become socially inappropriate against their will. PMID- 16248708 TI - Hierarchical linear modeling analyses of the NEO-PI-R scales in the Baltimore Longitudinal Study of Aging. AB - The authors examined age trends in the 5 factors and 30 facets assessed by the Revised NEO Personality Inventory in Baltimore Longitudinal Study of Aging data (N=1,944; 5,027 assessments) collected between 1989 and 2004. Consistent with cross-sectional results, hierarchical linear modeling analyses showed gradual personality changes in adulthood: a decline in Neuroticism up to age 80, stability and then decline in Extraversion, decline in Openness, increase in Agreeableness, and increase in Conscientiousness up to age 70. Some facets showed different curves from the factor they define. Birth cohort effects were modest, and there were no consistent Gender x Age interactions. Significant nonnormative changes were found for all 5 factors; they were not explained by attrition but might be due to genetic factors, disease, or life experience. PMID- 16248711 TI - Perceptions of and satisfaction with retirement: a comparison of six European union countries. AB - The main aim of this study was to assess perceptions of and satisfaction with retirement among 1,686 recently retired people from 6 European Union countries. The authors compared their responses to a set of questions taken from the Retirement Satisfaction Inventory (F. J. Floyd et al., 1992). The first significant result was that Belgian, British, Finnish, French, and Spanish retirees all expressed very similar assessments of life satisfaction, whereas Portuguese retirees expressed a lower level of satisfaction. The 2nd result was that the major determinants of overall satisfaction in the 6 countries were (a) health and resources and (b) anticipated satisfaction. Finally, the data revealed that cultural background only accounted for a limited amount of variance in overall satisfaction with retirement. PMID- 16248712 TI - The relation between source memory and episodic memory: comment on siedlecki et Al. (2005). AB - On the basis of an interesting structural equation analysis, K. L. Siedlecki, T. A. Salthouse, and D. E. Berish argued that "it may not be meaningful to refer to source memory as a construct distinct from episodic memory" (p. 31). This commentary highlights that this same point could also be made on conceptual grounds. To suggest that source and episodic memory are distinct concepts would confound tasks with theoretical constructs. All episodic tasks involve making attributions about the origin of mental experiences (source monitoring). Conversely, source memory tasks are designed to investigate episodic memory. No task is special, but each may be useful, depending on the focus of interest. PMID- 16248714 TI - An inkblot for attitudes: affect misattribution as implicit measurement. AB - Misattributions people make about their own affective reactions can be used to measure attitudes implicitly. Combining the logic of projective tests with advances in priming research, the affect misattribution procedure (AMP) was sensitive to normatively favorable and unfavorable evaluations (Experiments 1-4), and the misattribution effect was strong at both fast and slow presentation rates (Experiments 3 and 4). Providing further evidence of validity, the AMP was strongly related to individual differences in self-reported political attitudes and voting intentions (Experiment 5). In the socially sensitive domain of racial attitudes, the AMP showed in-group bias for Black and White participants. AMP performance correlated with explicit racial attitudes, a relationship that was moderated by motivations to control prejudice (Experiment 6). Across studies, the task was unaffected by direct warnings to avoid bias. Advantages of the AMP include large effect sizes, high reliability, ease of use, and resistance to correction attempts. PMID- 16248713 TI - Audience-tuning effects on memory: the role of shared reality. AB - After tuning to an audience, communicators' own memories for the topic often reflect the biased view expressed in their messages. Three studies examined explanations for this bias. Memories for a target person were biased when feedback signaled the audience's successful identification of the target but not after failed identification (Experiment 1). Whereas communicators tuning to an in group audience exhibited the bias, communicators tuning to an out-group audience did not (Experiment 2). These differences did not depend on communicators' mood but were mediated by communicators' trust in their audience's judgment about other people (Experiments 2 and 3). Message and memory were more closely associated for high than for low trusters. Apparently, audience-tuning effects depend on the communicators' experience of a shared reality. PMID- 16248715 TI - On the cultural guises of cognitive dissonance: the case of easterners and westerners. AB - Cognitive dissonance and effects of self-affirmation on dissonance arousal were examined cross-culturally. In Studies 1 and 2, European Canadians justified their choices more when they made them for themselves, whereas Asian Canadians (Study 1) or Japanese (Study 2) justified their choices more when they made them for a friend. In Study 3, an interdependent self-affirmation reduced dissonance for Asian Canadians but not for European Canadians. In Study 4, when Asian Canadians made choices for a friend, an independent self-affirmation reduced dissonance for bicultural Asian Canadians but not for monocultural Asian Canadians. These studies demonstrate that both Easterners and Westerners can experience dissonance, but culture shapes the situations in which dissonance is aroused and reduced. Implications of these cultural differences for theories of cognitive dissonance and self-affirmation are discussed. PMID- 16248716 TI - When authorities' commands backfire: attributions about consensus and effects on deviant decision making. AB - The perception of consensus typically produces conformity, but specific attributional circumstances may produce deviance instead. Ironically, the command of an authority figure may create one such circumstance. Participants were presented with scenarios in which they had to make a choice between 2 options. Prior to their decision, they observed others all making a single choice. In some conditions, this consensus occurred following an authority's explicit command to make that choice. Results revealed the hypothesized effect--the authority's command led participants to make deviant decisions-and revealed that this effect was moderated by the authority's continued presence, expertise, the target(s) of the command, and the ability of perceivers to use their cognitive resources. PMID- 16248717 TI - Giving up and giving in: the costs and benefits of daily sacrifice in intimate relationships. AB - This research provided the first empirical investigation of how approach and avoidance motives for sacrifice in intimate relationships are associated with personal well-being and relationship quality. In Study 1, the nature of everyday sacrifices made by dating partners was examined, and a measure of approach and avoidance motives for sacrifice was developed. In Study 2, which was a 2-week daily experience study of college students in dating relationships, specific predictions from the theoretical model were tested and both longitudinal and dyadic components were included. Whereas approach motives for sacrifice were positively associated with personal well-being and relationship quality, avoidance motives for sacrifice were negatively associated with personal well being and relationship quality. Sacrificing for avoidance motives was particularly detrimental to the maintenance of relationships over time. Perceptions of a partner's motives for sacrifice were also associated with well being and relationship quality. Implications for the conceptualization of relationship maintenance processes along these 2 dimensions are discussed. PMID- 16248718 TI - The possessive self as a barrier to conflict resolution: effects of mere ownership, process accountability, and self-concept clarity on competitive cognitions and behavior. AB - The authors propose that people have difficulty managing conflict because they quickly develop ownership of arguments and positions they use in the dispute, that these arguments and positions become part of their (extended) self-concept, and that any opposition or counterargumentation therefore becomes an ego-threat. Four studies reveal that individuals value arguments and beliefs more when these are associated with the self and that anticipated or real opposition triggers ego defensive cognition and behavior, including competitive communication, retaliatory responses, negative perceptions of the partner, and attitude polarization. These effects were weaker when epistemic needs were raised through process accountability or when individuals had high rather than low self-concept clarity. The authors conclude that because people develop ownership of arguments and make these part of their self-concept, conflict is difficult to manage and bound to escalate. PMID- 16248719 TI - Conflict's consequences: effects of social motives on postnegotiation creative and convergent group functioning and performance. AB - Two studies tested the effects of social motives during negotiation on postnegotiation group performance. In both experiments, a prosocial or a proself motivation was induced, and participants negotiated in 3-person groups about a joint market. In Experiment 1, groups subsequently performed an advertisement task. Consistent with the authors' predictions, results showed that proself groups performed worse on the convergent aspects of this task but better on the divergent aspects than prosocial groups. In Experiment 2, the authors manipulated social motive and negotiation (negotiation vs. no negotiation), and groups performed a creativity task (requiring divergent performance) or a planning task (requiring convergent performance). Proself groups showed greater dedication, functioned more effectively, and performed better than prosocial groups on the creativity task, whereas prosocial groups showed greater dedication, functioned more effectively, and performed better than proself groups on the planning task, and these effects only occurred when the task was preceded by group negotiation. PMID- 16248720 TI - Responses to interpersonal transgressions in families: forgivingness, forgivability, and relationship-specific effects. AB - Social relations analyses examined the relative importance of forgivingness (disposition to forgive others), forgivability (tendency to obtain forgiveness from others), and relationship effects in determining family members' transgression-related interpersonal motivations (TRIMs) and their perceptions of others' TRIMs toward them (PTRIMs). In 2 studies, the individual and dyadic predictors of these components and their relative importance differed by family role (father, mother, or early adolescent child). Dispositional tendencies accounted for the most variance in father and child forgiveness, whereas mothers' TRIMs and PTRIMs were more strongly determined by relationship and partner effects. Personality correlates of forgivingness and forgivability were moderated by family role. The findings point to the need to embed the study of forgiveness in more complex psychosocial contexts. The theoretical, methodological, and applied implications of this conclusion are discussed. PMID- 16248721 TI - The influence of chronically and temporarily accessible information on life satisfaction judgments. AB - The authors examined the influence of temporarily and chronically accessible information on life satisfaction judgments. Meta-analyses revealed high retest reliability of life satisfaction judgments and weak effects of the item order of domain and global satisfaction judgments. Study 1 (N=225) failed to replicate a widely cited finding of strong item-order effects. In Studies 2 (N=100), 3 (N=200), and 4 (N=222), chronically accessible information was a strong predictor of life satisfaction judgments, whereas item order had a relatively small effect. Study 5 (N=651) demonstrated that the results generalize to single item measures and judgments of shorter time periods. The results suggest that life satisfaction judgments are more heavily based on chronically accessible than temporarily accessible information. PMID- 16248722 TI - Personality profiles of cultures: aggregate personality traits. AB - Personality profiles of cultures can be operationalized as the mean trait levels of culture members. College students from 51 cultures rated an individual from their country whom they knew well (N=12,156). Aggregate scores on Revised NEO Personality Inventory (NEO-PI-R) scales generalized across age and sex groups, approximated the individual-level 5-factor model, and correlated with aggregate self-report personality scores and other culture-level variables. Results were not attributable to national differences in economic development or to acquiescence. Geographical differences in scale variances and mean levels were replicated, with Europeans and Americans generally scoring higher in Extraversion than Asians and Africans. Findings support the rough scalar equivalence of NEO-PI R factors and facets across cultures and suggest that aggregate personality profiles provide insight into cultural differences. PMID- 16248723 TI - Intention memory and achievement motivation: volitional facilitation and inhibition as a function of affective contents of need-related stimuli. AB - Replicating findings of J. Kuhl and M. Kazen (1999), reduction or removal of Stroop interference was achieved after short exposure to primes eliciting positive affect. This effect was need specific: Stroop interference removal (volitional facilitation) was found with positive primes related to achievement needs but not with positive primes related to affiliation or power needs. Five studies are reported. College students and unemployed university graduates participated in 2 studies each and branch managers of a large insurance company in 1 study. Whereas Stroop interference reduction or removal was found in all groups after positive-achievement primes, the 2 groups of unemployed persons additionally showed a significant increase of Stroop interference (volitional inhibition) after exposure to primes related to negative achievement episodes. Results are discussed in the context of Kuhl's personality systems interactions theory. PMID- 16248724 TI - Don't worry, be happy? Neuroticism, trait-consistent affect regulation, and performance. AB - People regulate their affect either to feel good or to achieve instrumental success. The present experiments show that when driven by performance goals, people can be motivated to experience unpleasant affect when it is trait consistent, because of its instrumental benefits (e.g., M. Tamir & M. D. Robinson, 2004). In 4 studies, individuals high in neuroticism were more likely than those low in neuroticism to choose to increase their level of worry, as indicated by self-reported preferences (Study 1) and by behavioral choices in experimental settings (Studies 2-4). As predicted, such preferences were evident when expecting to perform demanding tasks but not when expecting an undemanding task (Study 2). Study 4 suggests that such preferences for short-term unpleasant affect may be beneficial to performance. PMID- 16248725 TI - What perceptual rules do capuchin monkeys (Cebus apella) follow in completing partly occluded figures? AB - Capuchin monkeys (Cebus apella) matched a variety of rodlike figures, distinguishable only by their central portions. The monkeys were then tested for perceptual completion rules by examining which comparison figures they would choose as matching the sample with the critical central portion occluded by a horizontal belt. In Experiment 1, the monkeys overwhelmingly chose a straight rod over disconnected rods and rods having irregular shapes at their center, irrespective of the presence-absence of common motion of visible parts of the samples. The monkeys chose a connected rod for relatable rods and disconnected rods for non-relatable ones in Experiment 2. In Experiments 3 and 4, some of the monkeys chose rods specified by the global regularity of the contour, whereas others did not. Experiment 5 showed that humans and capuchin monkeys basically follow similar perceptual rules in completing occluded contours, but the global regularity rule may be stronger in humans. PMID- 16248726 TI - Auditory--visual cross-modal perception of communicative stimuli in tufted capuchin monkeys (Cebus apella). AB - Research on cross-modal performance in nonhuman primates is limited to a small number of sensory modalities and testing methods. To broaden the scope of this research, the authors tested capuchin monkeys (Cebus apella) for a seldom-studied cross-modal capacity in nonhuman primates, auditory-visual recognition. Monkeys were simultaneously played 2 video recordings of a face producing different vocalizations and a sound recording of 1 of the vocalizations. Stimulus sets varied from naturally occurring conspecific vocalizations to experimentally controlled human speech stimuli. The authors found that monkeys preferred to view face recordings that matched presented vocal stimuli. Their preference did not differ significantly across stimulus species or other stimulus features. However, the reliability of the latter set of results may have been limited by sample size. From these results, the authors concluded that capuchin monkeys exhibit auditory-visual cross-modal perception of conspecific vocalizations. PMID- 16248727 TI - Interactions between conditioned and unconditioned flavor preferences. AB - Five experiments investigated how rats' conditioned preferences or aversions for aqueous odors paired with sucrose or salt are affected by their unconditioned response to those tastes. Rats preferred an odor paired with 30% sucrose over an odor paired with 5% sucrose when both were presented in 5% sucrose, but they showed no preference or, if thirsty, showed the reverse preference, when the odors were presented in 30% sucrose. These changes in conditioned preference corresponded to changes in the rats' unconditioned preference for the accompanying sucrose solution. Rats' conditioned aversions for odors paired with salt showed a similar dependence on their reaction to the accompanying salt solution. The results were interpreted as showing that conditioned and unconditioned flavor preferences combine additively, as if mediated by the same sensory representation. PMID- 16248728 TI - Inference-based retrospective revaluation in human causal judgments requires knowledge of within-compound relationships. AB - In an allergist causal-judgment task, food compounds were followed by an allergic reaction (e.g., AB+), and then 1 cue (A) was revalued. Experiment 1, in which participants who were instructed that whatever was true about one element of a causal compound was also true of the other, showed a reverse of the standard retrospective revaluation effect. That is, ratings of B were higher when A was causal (A+) than when A was safe (A-). This effect was taken to reflect inferential reasoning, not an associative mechanism. In Experiment 2, within compound associations were found to be necessary to produce this inference-based revaluation. Therefore, evidence that within-compound associations are necessary for retrospective revaluation is consistent with the inferential account of causal judgments. PMID- 16248729 TI - Learning processes in matching and oddity: the oddity preference effect and sample reinforcement. AB - Eight pigeons learned either matching (to sample) or oddity (from sample) with or without reward for sample responding. The training stimuli were coarse-white, fine-black, or smooth-mauve gravels in pots with buried grain as the reinforcer. Oddity without sample reward was learned most rapidly, followed by matching with sample reward, oddity with sample reward, and matching without sample reward. Transfer was related to acquisition rate: The oddity group without sample reward showed full (equal to baseline) color and texture transfer; the matching group with sample reward showed partial texture transfer; other groups showed no transfer. Sample reward was shown to determine rate of acquisition of matching and oddity and the oddity preference effect. The results are discussed in terms of item-specific associations operating early in learning prior to any relational learning between sample and comparison stimuli. PMID- 16248730 TI - The discrimination of structure: III. Representation of spatial relationships. AB - Pigeons received a discrimination in which the spatial relationship between 2 adjacent rectangles filled with different colors signaled the trial outcome. Test trials then involved the same rectangles separated horizontally by a gap. The tests in Experiment 1 disrupted the discrimination more when the rectangles were tall and thin than when they were short and wide. Experiment 2 revealed that the width of the rectangles rather than their height determined the extent to which separating them would disrupt the original discrimination. The results are explained in terms of a template-matching account of pattern recognition with the additional assumption, supported by Experiment 3, that the size of a template can be altered to improve its match with a test pattern. PMID- 16248731 TI - Control of matching by differential outcome expectancies in the absence of differential sample-outcome associations: a serial compound view. AB - In this study, pigeons learned 2 separate one-to-many conditional discriminations in which they matched form samples to line and hue comparisons. Correct choices within each comparison dimension yielded differential (food vs. no-food) outcomes that were not predictable from the samples alone. At asymptote, latency to make a correct choice was shorter when food was the contingent outcome than when no food was the outcome. More important, when the samples from each task were subsequently exchanged, comparison choice varied systematically as a function of the sample and the set of new comparison alternatives that followed them. Together, these results indicate that choices were cued by differential outcome expectancies arising from serial compounds consisting of each sample and the dimensional characteristics of the comparisons. PMID- 16248732 TI - Recovery from blocking between outcomes. AB - Contemporary associative learning research largely focuses on cue competition phenomena that occur when 2 cues are paired with a common outcome. Little research has been conducted to investigate similar phenomena occurring when a single cue is trained with 2 outcomes. Three conditioned lick suppression experiments with rats assessed whether treatments known to alleviate blocking between cues would also attenuate blocking between outcomes. In Experiment 1, conditioned responding recovered from blocking between outcomes when a long retention interval was interposed between training and testing. Experiment 2 obtained recovery from blocking between outcomes when the blocking outcome was extinguished after the blocking treatment. In Experiment 3, a recovery from blocking between outcomes occurred when a reminder stimulus was presented in a novel context prior to testing. Collectively, these studies demonstrate that blocking of outcomes, like blocking of cues, appears to be caused by a deficit in the expression of an acquired association. PMID- 16248733 TI - The experimental task influences cue competition in human causal learning. AB - Participants were shown A+ and C- trials followed by AB+ and CD+ trials. These trials were embedded in a causal learning task in which participants had to learn either the relationship between different foods and allergic reactions or the relationship between different stocks and an increase in the stock market index. The authors orthogonally varied the manner in which the different cues were presented to participants during training. Cue competition was related to the causal learning scenario but not to the manner in which the different cues were presented. These results question claims of a human bias toward configural processing that were based on difficulties in finding cue competition in some previous causal learning experiments. PMID- 16248734 TI - Interval timing with gaps: gap ambiguity as an alternative to temporal decay. AB - C. V. Buhusi, D. Perera, and W. H. Meck (2005) proposed a hypothesis of timing in rats to account for the results of experiments that have used the peak procedure with gaps. According to this hypothesis, the introduction of a gap causes the animal's memory for the pregap interval to passively decay (subjectively shorten) in direct proportion to the duration and salience of the gap. Thus, animals should pause with short, nonsalient gaps but should reset their clock with longer, salient gaps. The present authors suggest that the ambiguity of the gap (i.e., the similarity between the gap and the intertrial interval in both appearance and relative duration) causes the animal to actively reset the clock and prevents adequate assessments of the fate of timed intervals prior to the gap. Furthermore, when the intertrial interval is discriminable from the gap, the evidence suggests that timed intervals prior to the gap are not lost but are retained in memory. PMID- 16248735 TI - Reinforcer devaluation in palatability-based learned flavor preferences. AB - Rats exposed to simultaneous compounds of 1 neutral flavor with dilute (2%) sucrose and a 2nd flavor with dilute (2%) maltodextrin subsequently consumed both flavors in preference to a 3rd flavor that was never paired with a palatable taste. Brief training exposure under ad lib food and water minimized the post ingestive effects of nutrients, emphasizing the contribution of palatability to these preferences. Devaluation of sucrose or maltodextrin by pairing with illness (Experiment 1) or sensory-specific satiety (Experiment 2) selectively reduced the preference for the flavor previously paired with the devalued reinforcer. Such reinforcer-specific devaluation effects suggest that palatability-based learned flavor preferences are under-pinned by a Pavlovian process whereby the cue flavor is associated with the taste of the concurrently consumed palatable reinforcer. PMID- 16248736 TI - Thirty-something categorization results explained: selective attention, eyetracking, and models of category learning. AB - An eyetracking study testing D. L. Medin and M. M. Schaffer's (1978) 5-4 category structure was conducted. Over 30 studies have shown that the exemplar-based generalized context model (GCM) usually provides a better quantitative account of 5-4 learning data as compared with the prototype model. However, J. D. Smith and J. P. Minda (2000) argued that the GCM is a psychologically implausible account of 5-4 learning because it implies suboptimal attention weights. To test this claim, the authors recorded undergraduates' eye movements while the students learned the 5-4 category structure. Eye fixations matched the attention weights estimated by the GCM but not those of the prototype model. This result confirms that the GCM is a realistic model of the processes involved in learning the 5-4 structure and that learners do not always optimize attention, as commonly supposed. The conditions under which learners are likely to optimize attention during category learning are discussed. PMID- 16248737 TI - Eye gaze and individual differences consistent with learned attention in associative blocking and highlighting. AB - The associative learning effects called blocking and highlighting have previously been explained by covert learned attention, but evidence for learned attention has been indirect, via models of response choice. The present research reports results from eye tracking consistent with the attentional hypothesis: Gaze duration is diminished for blocked cues and augmented for highlighted cues. If degree of attentional learning varies across individuals but is relatively stable within individuals, then the magnitude of blocking and highlighting should covary across individuals. This predicted correlation is obtained for both choice and eye gaze. A connectionist model that implements attentional learning is shown to fit the data and account for individual differences by variation in its attentional parameters. PMID- 16248739 TI - Imageability and ambiguity effects in speeded naming: convergence and divergence. AB - A current area of controversy within the literature on visual word recognition concerns the extent to which semantic information influences the computation of phonology. Experiment 1 revealed that both the imageability effect and the ambiguity advantage seen in the speeded naming task are confined to words with atypical mappings between spelling and sound. Nonetheless, it is possible that either of these effects may arise from the operation of the direct rather than the semantic pathway. Experiment 2 therefore included nonword fillers in order to minimize semantic reliance during speeded naming. This manipulation removed the imageability effect, indicating a semantic locus, but the ambiguity advantage remained, suggesting a nonsemantic locus. These results are considered in the context of computational models that incorporate a semantic level of representation. PMID- 16248738 TI - Effects of contextual predictability and transitional probability on eye movements during reading. AB - In 2 eye-movement experiments, the authors tested whether transitional probability (the statistical likelihood that a word precedes or follows another word) affects reading times and whether this occurs independently from contextual predictability effects. Experiment 1 showed early effects of predictability, replicating S. A. McDonald and R. C. Shillcock's (2003a) finding that words with a high transitional probability (defeat following accept) are read faster than words with a low transitional probability (losses following accept). However, further analyses suggested that the transitional probability effect was likely due to differences in predictability rather than transitional probability. Experiment 2, using a better controlled set of items, again showed an effect of predictability, but no effect of transitional probability. The authors conclude that effects of transitional probability are part of regular predictability effects. Their data also show that predictability effects are detectable very early in the eye-movement record and between contexts that are weakly constraining. PMID- 16248740 TI - Constituent structure and linear order in language production: evidence from subject-verb agreement. AB - A number of studies have shown that structural factors play a much larger role than the linear order of words during the production of grammatical agreement. These findings have been used as evidence for a stage in the production process at which hierarchical relations between constituents have been established (a necessary precursor to agreement), but before the final linear order of words is determined. The current article combines evidence from off-line ratings, online production studies, and a corpus analysis in support of the view that linear order effects do exist. These findings have implications both for theories of agreement production and language production more generally. PMID- 16248741 TI - Context effects on lexical choice and lexical activation. AB - Speakers are regularly confronted with the choice among lexical alternatives when referring to objects, including basic-level names (e.g., car) and subordinate level names (e.g., Beetle). Which of these names is eventually selected often depends on contextual factors. The present article reports a series of picture word interference experiments that explored how the designated target name (basic level vs. subordinate level) and contextual constraints rendering the name alternative either appropriate or inappropriate affect lexical activation and lexical choice. The experimental data demonstrate clear context effects on the eventual lexical choice. However, they also show that alternative nonselected object names are phonologically activated, even if a constraining context makes these alternative names currently inappropriate. PMID- 16248742 TI - Semantic and phonological context effects in speech error repair. AB - When speakers repair speech errors, they plan the repair in the context of an abandoned word (the error) that is usually similar in meaning or form. Two picture-naming experiments tested whether the error's lexical representations influence repair planning. Context pictures were sometimes replaced with target pictures; the picture names were related in meaning or form or were unrelated. The authors measured target picture-naming latencies separately for trials in which the context name was interrupted or completed. Interrupted trials showed semantic interference and phonological facilitation, whereas completed trials showed semantic facilitation and phonological interference. Thus, errors influence repair production. The authors explain the polarity of these effects in terms of the literature on context effects in word production. PMID- 16248743 TI - Theoretical correlations and measured correlations: relating recognition and recall in four distributed memory models. AB - This article addresses the relation between item recognition and associative (cued) recall. Going beyond measures of performance on each task, the analysis focuses on the degree to which the contingency between successful recognition and successful recall of a studied item reflects the commonality of memory processes underlying the recognition and recall tasks. Specifically, 4 classes of distributed memory models are assessed for their ability to account for the relatively invariant correlation (approximately .5) between successive recognition and recall. Basic versions of each model either under- or overpredict the intertask correlation. Introducing variability in goodness-of-encoding and response criteria, as well as output encoding, enabled all 4 models to reproduce the moderate intertask correlation and the increase in correlation observed in 2 mixed-list experiments. This model-based analysis provides a general theoretical framework for interpreting contingencies between successive memory tests. PMID- 16248744 TI - Compelling untruths: content borrowing and vivid false memories. AB - False memories are sometimes accompanied by surprisingly vivid experiential detail that makes them difficult to distinguish from actual memories. Such strikingly real false memories may be produced by a process called content borrowing in which details from presented items are errantly borrowed to corroborate the occurrence of the false memory item. In 2 experiments using think out-loud protocols at both study and test, evidence for content borrowing occurred for more than half of the false remember judgments participants reported. The present study also provides evidence consistent with recollection rejection and distinctiveness playing a role in false-memory editing. PMID- 16248745 TI - The role of inhibitory control in the production of misinformation effects. AB - Recent research has indicated a link between retrieval-induced forgetting and the production of misinformation effects (J. Saunders & M. D. MacLeod, 2002). The mechanism underlying this relationship, however, remains unclear. In an attempt to clarify this issue, the authors presented 150 participants with misinformation under conditions designed to promote the activation of inhibitory control during the retrieval of information about a target event. A modified retrieval practice paradigm that used the independent probe method pioneered by M. C. Anderson and B. A. Spellman (1995) revealed that misinformation effects emerged only where misinformation had been introduced about items that had been subject to 1st order, 2nd-order, or cross-category inhibition. By contrast, misinformation effects failed to emerge where inhibitory processing had not been activated. These findings are discussed in terms of inhibitory control, memory malleability, and their implications for the interviewing of eyewitnesses. PMID- 16248746 TI - Encoding and retrieval during bimanual rhythmic coordination. AB - In 2 experiments, bimanual 1:1 rhythmic coordination was performed concurrently with encoding or retrieval of word lists. Effects of divided attention (DA) on coordination were indexed by changes in mean relative phase and recurrence measures of shared activity between the 2 limbs. Effects of DA on memory were indexed by deficits in recall relative to baseline. For DA at both encoding and retrieval, the equilibrium values of relative phase were shifted and the degree of shared activity between left and right rhythmic motions was reduced. Recall was reduced, however, only for DA at encoding. The results corroborate and extend those obtained with more conventional secondary tasks (e.g., visual reaction time), suggesting attention dissimilarities between episodic encoding and retrieval. PMID- 16248747 TI - Unconscious primes activate motor codes through semantics. AB - Today, it is generally accepted that unconscious stimuli can activate a response code, which leads to a response congruency effect (RCE) on a subsequent target. However, it is not yet clear whether this is due to the semantic processing of the primes or to the formation of direct stimulus-response (S-R) associations bypassing the semantic system. Recently, it was shown that even novel primes, for which no direct S-R links exist, can also evoke an RCE that is in line with the activation of response codes through semantics. In these experiments, the authors examined 3 alternatives for this RCE from novel primes and report a novel effect in unconscious priming. First, the authors show that this effect is not limited to a small set of numerical stimuli but also extends to letter stimuli (Experiments 1-3). Second, the authors show that the RCE is not a side effect of the prime-target distance effect, as has been reported before (Experiments 1-2). Third, the authors found that, for RCE to occur, overlap at the motor level but not at the semantic level was crucial (Experiments 2-3). Finally, in addition, the results showed a category match priming effect independent of RCE. This last result is evidence that novel unconscious primes activate their semantic category prior to the target and might be considered a good marker for semantic processing. PMID- 16248748 TI - Selective and nonselective transfer: positive and negative priming in a multiple task environment. AB - Processing of a probe stimulus can be affected either positively or negatively by presenting a related stimulus immediately before it. According to structural accounts, such effects occur because processing of the prime activates or inhibits the mental representation of the probe before it is presented. In contrast, transfer-appropriate processing accounts suggest that success in processing a probe depends on resources made available by earlier experiences of related stimuli. The authors manipulated the similarity between the prime and probe on color, lexical status, and orthographic structure, requiring either lexical decision or color identification on each. The authors observed a complex pattern of positive and negative transfer that cannot easily be explained through activation-inhibition of mental structures. Instead, that pattern provides evidence in favor of transfer-appropriate processing. PMID- 16248749 TI - The influence of cue type on backward inhibition. AB - Backward inhibition is proposed as a process of lateral inhibition that operates during response selection in task switching, reducing interference caused by the most recently abandoned task set. The effect has been observed across a wide range of contexts but is eliminated by using spatial location to cue tasks (K. D. Arbuthnott & T. S. Woodward, 2002). The present studies replicated this finding, showing that spatial cues are also associated with greater response congruity than verbal cues, consistent with the lateral inhibition model. Spatial cues may introduce greater discriminability between competing category-response rules, reducing the need for lateral inhibition. However, when participants named the task before target presentation, backward inhibition was observed with spatial cues, suggesting that verbalization increased competition between sets, despite spatial localization. PMID- 16248750 TI - Selective interference with the use of visual images in the symbolic distance paradigm. AB - Eight experiments investigated the effects of visual, spatial, auditory, and executive interference on the symbolic comparison of animal size and ferocity, semantic goodness of words, and numbers. Dynamic visual noise (DVN) and the reading of visually presented stimulus items were shown to selectively interfere with response times on the animal size comparison task, though the slope of the symbolic distance function remained unchanged. Increased change of DVN significantly increased interference, but interference was reduced by equiluminant DVN. Spatial tracking reduced the slope of the symbolic distance function in contrast to an executive task that only increased mean latency and errors for all comparisons. Results suggest that the generation of an image is necessary for size comparison, but neither imagery nor executive function is responsible for the frequently observed distance-time function. PMID- 16248751 TI - Allocentric coding of object-to-object relations in overlearned and novel environments. AB - R. F. Wang and E. S. Spelke's (2000) finding that disorientation disrupts knowledge is consistent with egocentric but not allocentric coding of object location. The present experiments tested the hypothesis that egocentric coding may dominate early on but that once an allocentric representation is established, then target location is retrieved from it. This hypothesis predicts that disorientation will disrupt configuration knowledge in a novel environment, such as that used by Wang and Spelke, but not in an overlearned environment. Experiments 1 and 2 tested whether disorientation disrupted configuration knowledge of an overlearned environment, and Experiments 3-7 tested whether disorientation disrupted configuration knowledge of a novel, room-sized environment. In none of the experiments did disorientation disrupt configuration knowledge. Hence, in addition to showing allocentric coding of overlearned interlandmark relations, the present findings are consistent with the immediate availability of allocentric location codes in a novel, room-sized environment. PMID- 16248752 TI - Evaluating three criteria for establishing cue-search hierarchies in inferential judgment. AB - The authors identify and provide an integration of 3 criteria for establishing cue-search hierarchies in inferential judgment. Cues can be ranked by information value according to expected information gain (Bayesian criterion), cue-outcome correlation (correlational criterion), or ecological validity (accuracy criterion). All criteria significantly predicted information acquisition behavior; however, in 3 experiments, the most successful predictor was the correlational criterion (followed by the Bayesian). Although participants showed sensitivity to task constraints, searching for less information when it was more expensive (Experiment 1) and when under time constraints (Experiment 2), concomitant changes in the relative frequency of acquisition of cues with different information values were not observed. A rational analysis illustrates why such changes in the frequency of acquisition would be beneficial, and reasons for the failure to observe such behavior are discussed. PMID- 16248753 TI - Dissociation between judgments and outcome-expectancy measures in covariation learning: a signal detection theory approach. AB - A number of studies using trial-by-trial learning tasks have shown that judgments of covariation between a cue c and an outcome o deviate from normative metrics. Parameters based on trial-by-trial predictions were estimated from signal detection theory (SDT) in a standard causal learning task. Results showed that manipulations of P(c) when contingency (deltaP) was held constant did not affect participants' ability to predict the appearance of the outcome (d') but had a significant effect on response criterion (c) and numerical causal judgments. The association between criterion c and judgment was further demonstrated in 2 experiments in which the criterion was directly manipulated by linking payoffs to the predictive responses made by learners. In all cases, the more liberal the criterion c was, the higher judgments were. The results imply that the mechanisms underlying the elaboration of judgments and those involved in the elaboration of predictive responses are partially dissociable. PMID- 16248754 TI - Temporal discounting when the choice is between two delayed rewards. AB - The present experiments extend the temporal discounting paradigm from choice between an immediate and a delayed reward to choice between 2 delayed rewards: a smaller amount of money available sooner and a larger amount available later. Across different amounts and delays, the data were consistently well described by a hyperbola-like discounting function, and the degree of discounting decreased systematically as the delay to the sooner reward increased. Three theoretical models (the elimination-by-aspects, present-value comparison, and common-aspect attenuation hypotheses) were evaluated. The best account of the data was provided by the common-aspect attenuation hypothesis, according to which the common aspect of the choice alternatives (i.e., the time until the sooner reward is available) receives less weight in the decision-making process. PMID- 16248755 TI - Following the wrong footsteps: fixation effects of pictorial examples in a design problem-solving task. AB - Two experiments examined possible negative transfer in nonexperts from the use of pictorial examples in a laboratory design problem-solving situation. In Experiment 1, 89 participants were instructed to "think aloud" and were assigned to 1 of 3 conditions: (a) control (standard instructions), (b) fixation (inclusion of a problematic example, describing its problematic elements), or (c) defixation (inclusion of a problematic example, with instructions to avoid using problematic elements). Negative transfer due to examples was measured both quantitatively and qualitatively through verbal protocols. Verbal protocols (N = 176) were analyzed for participants' reasons for reference to the examples. In Experiment 2, fixation to examples was evaluated in nonverbalizing participants (N = 60). Results of both experiments suggest that (a) although participants consulted the problem instructions, they tended to follow the examples even when they included inappropriate elements and (b) the fixation effects can be diminished with the use of defixating instructions. PMID- 16248757 TI - Counterfactual and semifactual conditionals prime alternative possibilities. AB - The authors examined in 3 experiments the comprehension of counterfactuals, such as "If it had rained, the plants would have bloomed," and semifactuals, such as "Even if it had rained, the plants would have bloomed," compared with indicative conditionals, "If it rained, the plants bloomed." The first experiment showed that people read the negative conjunction, "not p and not q" faster when it was primed by a counterfactual than when it was primed by an indicative conditional. They read the affirmative conjunction, "p and q" equally quickly when it was primed by either conditional. The 2nd experiment showed that people read the negated-antecedent conjunction, "not p and q" faster when it was primed by a semifactual conditional. The 3rd experiment corroborated these results in a direct comparison of counterfactuals and semifactuals. The authors discuss the implications of the results for the mental representations of different conditionals. PMID- 16248758 TI - The positive and negative consequences of multiple-choice testing. AB - Multiple-choice tests are commonly used in educational settings but with unknown effects on students' knowledge. The authors examined the consequences of taking a multiple-choice test on a later general knowledge test in which students were warned not to guess. A large positive testing effect was obtained: Prior testing of facts aided final cued-recall performance. However, prior testing also had negative consequences. Prior reading of a greater number of multiple-choice lures decreased the positive testing effect and increased production of multiple-choice lures as incorrect answers on the final test. Multiple-choice testing may inadvertently lead to the creation of false knowledge. PMID- 16248759 TI - An observation on the role of context variability in free recall. AB - The authors conducted 3 experiments investigating the effect of context variability and word frequency on free recall. Context variability refers to the number of pre-experimental contexts in which a given word is experienced. Both between-subjects and within-subjects manipulations of context variability demonstrated a distinct advantage for low context variability words. Standard word frequency effects were obtained in 2 of the 3 experiments, but the common finding of no word frequency differences in mixed lists of high and low word frequency may depend on the level (low vs. high) of context variability. The authors speculate that the advantage for low context variability items may accrue from better item-to-list context associations or better storage of contextual information as a consequence of the smaller pre-experimental contextual fan that these items possess. PMID- 16248761 TI - Knowledge and behaviour of tourists towards the sun, as studied in a region of northern Greece. AB - INTRODUCTION: Melanoma is considered one of the most malignant cancers. Its appearance is related to various factors such as ultraviolet radiation, recurrent sunburn, and phototype. During the summer holidays approximately 14 million tourists from northern European countries visit Greece; this does not include the local tourists. AIMS: To discover the extent of European visitors' awareness of the risk of sunburn. To explore the level of knowledge gained by local and foreign tourists from preventive campaigns related to the harmful effects of sun exposure. PARTICIPANTS: The sample consisted of 802 travelers from northern European countries, and 726 Greeks who departed from the airport 'Megas Alexandros' during August and September 2002. METHODS: A structured questionnaire was used to gain data about demographics, family history of skin cancer, and identification of phototype. Also requested was information about the hours of sun exposure, and the use of sunscreen (the pattern of application and its sun protective factor [SPF]). Comparisons of mean values between groups were made by Student's t-test, and the association between categorical variables was tested by Pearson's chi(2). Regression analysis was performed to examine the influence of different factors on the likelihood of being sunburned. RESULTS: The majority of foreigners belonged to phototype II or III, while the Greek tourists belonged to phototype III. Foreigners had a higher tendency to burn and a lower tendency to tan. Of the total sample, women presented a higher prevalence of sunburn (p <0.05), and they used sunscreen more frequently (p <0.001) than men did (p <0.001), in both groups. The mean SPF used was 17.3 (SD = 8.98) for the foreigners and 16.0 (SD = 2.0) for the Greeks. The media was the main source of information for both groups. Multiple regression analysis showed that skin type was significantly associated with a high probability of sunburn (p <0.001). Moreover, freckles (p <0.05) were a predictive factor for future sunburn. CONCLUSIONS: Our study showed that skin type is the most important predictor of future sunburn. The role of the primary care physician is to promote and encourage healthy habits, including attitude towards sun exposure. Increased susceptibility to sun-induced damage of persons with phototypes I and II mandates their identification as a target group in all media campaigns. PMID- 16248764 TI - Continuous infusion interleukin-2 and antihistamines in metastatic kidney cancer. AB - A prior randomized trial suggested a possible survival advantage favoring the combination of histamine and subcutaneous interleukin-2 (IL-2), compared to IL-2 alone in patients with metastatic melanoma. It has been postulated previously that antihistamines may, therefore, actually be antagonistic to IL-2 and thus interfere with its antitumor activity. We have previously shown no such antagonistic effect in patients with melanoma receiving IL-2 and antihistamines when reviewing the known literature. We sought to determine whether there was any negative effect of the combination in patients with metastatic kidney cancer. A PubMed literature search between 1985 and 2005 was done. High-dose continuous (or constant) infusion (CIV) interleukin-2 was used as the reference therapy because of the relatively constant IL-2 levels generated by this approach. Studies in which cimetidine, ranitidine, or famotidine were regularly scheduled and administered concurrently with IL-2 were included. Thirteen studies were identified. A total of 47 patients responded to therapy. Total response rate = 22%; 95%; Confidence Interval: 17%-28%. Eleven complete responses were noted. Complete response rate = 5%; 95% Confidence Interval: 3%-9%. These response rates are consistent with previously noted IL-2 response rates. In this study of CIV IL 2 and antihistamines, this combination appears to be active in metastatic kidney cancer. There appears to be no negative effect of antihistamine on the CIV IL-2 response rate in this disease. PMID- 16248763 TI - A retrospective study of biochemotherapy for metastatic melanoma: the importance of dose intensity. AB - BACKGROUND: The utility of biochemotherapy for metastatic melanoma remains controversial. Dose intensity has been recognized as an important determinant of response and survival in the chemotherapy of several malignancies but has not been studied in biochemotherapy. In this retrospective study, we described the relationship between achieved dose intensity and the response rate of inpatient decrescendo biochemotherapy at our center. METHODS: A study of 38 consecutive patients with metastatic melanoma was undertaken. The planned doses were dacarbazine 800 mg/m(2) on day 1 or temozolomide 150 mg/m(2) on days 1-4, cisplatin 20 mg/m(2) on days 1-4, vinblastine 1.5 mg/m(2) on days 1-4, interferon alpha-2b (Schering) 5 million IU (MIU)/m(2) on days 1-5, and interleukin-2 36 MIU on day 1, 18 MIU on day 2, and 9 MIU on days 3 and 4. RESULTS: Of 38 patients that received a total of 204 cycles of therapy, 8 (21%) complete responses and 14 (37%) partial responses were observed for an objective response rate of 58%. Median survival was 19.6 months. Achieved dose intensity was high with patients receiving 98.7% interleukin- 2, 87.1% interferon, 90.7% dacarbazine (DTIC), 94% temozolomide, 87.2% cisplatin, and 89.7% vinblastine. CONCLUSIONS: Six cycles of inpatient decrescendo biochemotherapy can be given with high-dose intensity and acceptable toxicity. High response rates with biochemotherapy for melanoma may correlate with dose intensity, dose density, and the number of cycles given on time. PMID- 16248762 TI - Phase I study of 90Y-CC49 monoclonal antibody therapy in patients with advanced non-small cell lung cancer: effect of chelating agents and paclitaxel co administration. AB - PURPOSE: This trial was designed to evaluate strategies to improve the efficacy of a radiolabeled monoclonal antibody (mCC49) against tumor-associated glycoprotein-72 (TAG-72) in patients with non-small cell lung cancer (NSCLC). The aims of this study were to determine: safety and maximum tolerated dose (MTD) of (90)Y-mCC49 in combination with interferon alpha2beta (IFN); whether calcium disodium versonate (EDTA) or diethylenetriamine penta-acetic acid (DTPA) could reduce myelosuppression; and safety and MTD of paclitaxel (Taxol) in combination with (90)Y-mCC49. EXPERIMENTAL DESIGN: Patients with advanced (TAG-72 positive) non-small cell lung cancer were entered in three phases; the first was the dose escalation of a single agent (90)Y-mCC49. In the second phase, the dose escalation of (90)Y-mCC49 was attempted with concurrent EDTA or DTPA chelator therapy. In the third phase, radiosensitization with a continuous infusion of paclitaxel (96-hour) was administered with (90)Y-mCC49. All patients received IFN for TAG-72 up-regulation. RESULTS: Thirty-four patients were evaluable. Reversible Grade 4 neutropenia and thrombocytopenia were the dose-limiting toxicities (DLTs). The MTD of (90)Y-mCC49/IFN was 14 mCi/m(2). EDTA did not alter toxicity, while there was a modest reduction of myelosuppression with DTPA. The MTD of continuous infusion paclitaxel in combination with 14 mCi/m(2) of (90)Y CC49 was 60 mg/m(2). There were no objective tumor responses. CONCLUSIONS: (90)Y mCC49/IFN was well tolerated at a dose of 14 mCi/m(2). The clinical effect of adjunctive chelating therapy with DTPA was modest. The MTD of coadministered continuous infusion (96-hour) paclitaxel was 60 mg/m(2). Because of the immunogenicity of the murine compound, future studies are planned using a humanized version of CC49. PMID- 16248765 TI - Strategies of antigen-specific T-cell-based immunotherapy for cancer. AB - The critical role of antigen-specific T-cells in the eradication of cancer has been demonstrated in numerous animal models, while significant challenges need to be conquered before antigen-specific T-cell immunotherapy can achieve true success in clinical practice. These challenges include: (1) weak or nonimmunogenicity of spontaneous tumors, (2) negative immune regulation mechanisms of the host immune system, (3) immune inhibition exerted by tumor cells, (4) physical barrier in solid tumor, and (5) escape or resistance to immune attack by tumor cells. Nonetheless, significant success has been achieved in several clinical trials recently, highlighting the possibility of successful manipulation of the immune system for control and elimination of tumor. We focused our study on summarizing the current knowledge and corresponding strategies for improving autologous cytotoxic T-cell (CTL)-based cancer immunotherapy, which include the following aspects: (1) the selection of tumor antigens for stimulation of CTL, (2) strategies of enhancing maturation and antigen presentation activity of dendritic cells (DC), (3) strategies of activation and maintenance of CTL response, and (4) recruitment of suitable immune effector cells to tumor sites. The successful manipulation of the immune system, based on the more and more detailed knowledge of tumor immunology, may finally reach the goal of "immune surveillance of malignancy." PMID- 16248766 TI - Intraperitoneal radioimmunotherapy with a humanized anti-TAG-72 (CC49) antibody with a deleted CH2 region. AB - The application of intraperitoneal (i.p.) radioimmunotherapy to treat i.p. tumor loci has been limited by bone marrow toxicity secondary to circulating radiolabeled antibodies. The generation of novel genetically engineered monoclonal antibodies, which can achieve high tumor uptake and rapid blood clearance, should enhance the therapeutic index of i.p. radioimmunotherapy. In this regard, a novel humanized anti-TAG-72 monoclonal antibody with a deleted CH2 region (HuCC49DeltaCH2) has been described, which localized well to subcutaneous xenograft tumors and had a rapid plasma clearance. The aim of this study was to examine the characteristics of this radiolabeled reagent when administered through the i.p. route in mice bearing i.p. tumor (LS174T). The DeltaCH2 molecule and intact humanized CC49 (HuCC49) monoclonal antibody were conjugated to PA-DOTA and radiolabeled with (177)Lu. Both molecules retained high-affinity binding to TAG-72 positive LS174T tumor cells in vitro. The radiolabeled DeltaCH2 molecule had a modest decrease in tumor localization, as compared to the intact molecule when administered i.p. to tumor-bearing mice and a dramatically shorter plasma disappearance T(1/2) at 2.7 hours compared to 61.2 hours for the intact antibody. The radiolabeled DeltaCH2 molecule thus had very high tumor:blood ratios. Using an (131)I-labeled system, the maximum tolerated dose of DeltaCH2 was >3x that of intact HuCC49. Autoradiography of tumors showed low radiation dose rates at tumor centers early (1 and 4 hours), as compared to higher dose rates at tumor periphery but a more uniform distribution by 24 hours. Dose-rate distributions were similar for both reagents. Animals bearing LS174T i.p. tumors were treated with 300 microCi of (177)Lu-labeled DeltaCH2 or intact HuCC49 by i.p. route daily x 3. The (177)Lu-DeltaCH(2) molecule mediated an increase in median survival compared to controls (67.5 +/- 7.5 days versus controls of 32 +/- 3.3) while the same dose of (177)Lu-HuCC49 produced early toxic deaths. These studies suggest that i.p. radioimmunotherapy using radiolabeled HuCC49DeltaCH2 should allow higher radiation doses to be administered with less marrow toxicity and potentially improved efficacy. PMID- 16248767 TI - In vitro characterization of 211 At-labeled antibody A33--a potential therapeutic agent against metastatic colorectal carcinoma. AB - The humanized antibody A33 binds to the A33 antigen, expressed in 95% of primary and metastatic colorectal carcinomas. The restricted pattern of expression in normal tissue makes this antigen a possible target for radioimmunotherapy of colorectal micrometastases. In this study, the A33 antibody was labeled with the therapeutic nuclide (211)At using N-succinimidyl para-(tri-methylstannyl)benzoate (SPMB). The in vitro characteristics of the (211)At-benzoate-A33 conjugate ((211)At-A33) were investigated and found to be similar to those of (125)I benzoate-A33 ((125)I-A33) in different assays. Both conjugates bound with high affinity to SW1222 cells (K(d) = 1.7 +/- 0.2 nM, and 1.8 +/- 0.1 nM for (211)At A33 and (125)I-A33, respectively), and both showed good intracellular retention (70% of the radioactivity was still cell associated after 20 hours). The cytotoxic effect of (211)At-A33 was also confirmed. After incubation with (211)At A33, SW1222 cells had a survival of approximately 0.3% when exposed to some 150 decays per cell (DPC). The cytotoxic effect was found to be dose-dependent, as cells exposed to only 56 DPC had a survival of approximately 5%. The (211)At-A33 conjugate shows promise as a potential radioimmunotherapy agent for treatment of micrometastases originating from colorectal carcinoma. PMID- 16248768 TI - Identification of an antilaminin-1 scFv that preferentially homes to vascular solid tumors. AB - The tumor vasculature and extracellular matrix make attractive targets for distinguishing solid tumors from normal cells. In solid tumors, the processes of angiogenesis and metastasis potentially give rise to unique epitopes not usually accessible in homeostatic organs. Specific targeting of solid tumors for radioimmunotherapy requires that the targeting agent accumulate rapidly and at high levels at the tumor site. This study involved the selection of scFvs that recognize laminin-1 in vitro from the Tomlinson I and J phage display libraries. Selected, purified scFvs were radioiodinated and injected in tumor-bearing mice. One of these, scFv 15-9, exhibited preferential accumulation at subcutaneous tumors when compared to other antilaminin scFvs or to a control scFv. Autoradiographic analysis indicated that scFv15- 9 also displayed a higher vessel:parenchyma ratio than did two other antilaminin scFvs, scFv 15-6 and scFv 15-1, indicating a preferential accumulation of scFv 15-9 around vessel structures. Immunohistochemistry confirmed that scFv 15-9 accumulated at sites of endothelial cells lining vessel structures where significant levels of laminin were present. These data demonstrate that scFv 15-9 binds to a specific epitope on laminin and has potential for tumor endoradiotherapy in subcutaneous tumors. PMID- 16248769 TI - Radioimmunotargeting of human rhabdomyosarcoma using monoclonal antibody 8H9. AB - Although metastatic rhabdomyosarcoma (RMS) is chemotherapy and radiotherapy responsive, few patients are cured. 8H9, a murine IgG(1) monoclonal antibody, recognizes a unique cell surface tumor antigen broadly distributed on neuroectodermal, epithelial, and mesenchymal tumors, including RMS. We now report on the in vitro characterization of radiolabeled 8H9 and its in vivo immunotargeting potential in mice with subcutaneous human RMS. Saturation-binding studies carried out to determine (125)I-8H9 affinity to the RMS cell line HTB82 demonstrated that (125)I-8H9 had a K(d) of 10.3nM with an estimated 115,000 binding sites on every HTB82 cell. (125)I-8H9 was retained on the cell surface without significant internalization. Biodistribution of (125)I-8H9 was studied in athymic mice bearing HTB82 xenografts. Following intravenous injection of 4.44MBq of (125)I-8H9, selective tumor uptake was evident 4 to 172 hours after injection. Average tumor uptake was 7.0 +/- 1.8, 11.5 +/- 3.9, 15.1 +/- 3.7, and 5.4 +/- 1.2% injected dose per gram at 4, 24, 48, and 172 hours, respectively. Mean tumor: tissue ratios were maximal at 172 hours (for lung, 4, kidney, 6, liver, 7, spleen, 11, femur, 14, muscle, 18, and brain, 48). Established RMS xenografts treated with a single injection of 18.5 MBq (131)I-8H9 were significantly suppressed compared to controls. Radiolabeled 8H9 effectively targeted RMS xenografts and may have a potential clinical role in radioimmunotherapy. PMID- 16248770 TI - Convenient solid-phase synthesis of diethylenetriaminepenta-acetic acid (DTPA)- conjugated cyclic RGD peptide analogues. AB - Solid-phase synthesis of radiometal chelator-conjugated peptides can facilitate the creation of radioactive peptide libraries to be utilized in high throughput in vivo screening of targeted nuclear-imaging agents. In this study, a new diethylenetriaminepentaacetic acid (DTPA) derivative, 1-(p-succinamidobenzyl)- DTPA penta-t-butyl ester [DTPA(But)(5)-Bz-NH-SA], and its precursor molecule, 1 (p-aminobenzyl)- DTPA penta-t-butyl ester (DTPA(But)(5)-Bz-NH(2)), were applied to the solid-phase synthesis of DTPA-conjugated cyclic peptides containing the Arg-Gly-Asp (RGD) motif with high efficiency. The resulting conjugates, DTPA-Bz NH-SA-c(Lys-Arg-Gly-Asp-phe) [DTPA-Bz-NH-SA-c(KRGDf)] and DTPA-Bz-NHc( Glu-Arg Gly-Asp-phe) [DTPA-Bz-NH-c(KRGDf)], demonstrated similar in vitro biologic activities as their corresponding parent peptides. (111)In-labeled, DTPA conjugated RGD peptides showed selective binding to integrin alphavbeta3 in human melanoma M21 tumors grown in nude mice. Furthermore, (111)In-DTPABz- NH-c(ERGDf) showed lower retention in the liver and the kidney than (111)In-DTPA-Bz-NH-SAc( KRGDf) did, which contributed to higher target to nontarget ratio for (111)In DTPA-Bz-NH-c(ERGDf). The method reported here can be extended to the construction of peptide libraries containing DTPA for high throughput in vitro and in vivo screening of molecularly targeted imaging agents. PMID- 16248771 TI - Alpha-particle radioimmunotherapy of disseminated peritoneal disease using a (212)Pb-labeled radioimmunoconjugate targeting HER2. AB - These studies demonstrate the feasibility of targeted therapy for the treatment of disseminated peritoneal disease using (212)Pb-labeled Herceptin as an in vivo generator of (212)Bi. In vitro studies compare the potential of the bismuth radioisotopes, (213)Bi and (212)Bi, to that of (212)Pb. Overall, (212)Pb results in a higher therapeutic index than either bismuth radioisotope, requiring lower radioactivity (microCi) for effective cytotoxic response. A pilot radioimmunotherapy (RIT) experiment treating mice bearing 5 d LS-174T intraperitoneally (i.p.) xenografts determined a maximum tolerated dose (MTD) of 20-40 microCi with i.p. administration. A specific dose response was observed and 10 microCi was selected as the effective operating dose for future experiments. Median survival of tumor-bearing mice receiving 10 microCi increased from 19 to 56 days (p = 0.008). The efficacy of (212)Pb-Herceptin was also assessed in a human pancreatic carcinoma xenograft (Shaw; i.p.) animal model previously reported as unresponsive to 213Bi-Herceptin (p = 0.002). Multiple dosing of (212)Pb-Herceptin was evaluated in both animal models. The median survival of mice bearing 3 d LS-174T i.p. xenografts increased to 110 days, with up to 3 doses of (212)Pb-Herceptin given at approximately monthly intervals; however, there was no evidence of a correlation with the second and third doses (p = 0.98). No improvement in median survival was noted with a similar regimen in the Shaw xenograft model. PMID- 16248773 TI - Endothelin-1 and angiogenesis in cancer. AB - Tumours require oxygenation, nutrition and a route for dissemination. This necessitates the development of new vessels or angiogenesis. High levels of new vessel development are indicators of poor prognosis in cancer; they also provide new avenues of anti-tumour therapy. Angiogenesis in cancer produces structurally different vessels from angiogenesis in wound healing and inflammation. This article reviews the differences between vessels in tumour angiogenesis and normal angiogenesis. The main focus of the article is the role of the vasoactive peptide endothelin-1 (ET-1) in tumour angiogenesis. The role of ET-1 in tumour development is reviewed, before the direct and indirect effects of ET-1 in angiogenesis are examined. ET-1 has a direct angiogenic effect on endothelial and peri-vascular cells. It also has an indirect action through the increased release of the potent pro-angiogenic substance vascular endothelial growth factor (VEGF), via hypoxia inducible factor-1. ET-1 also indirectly stimulates angiogenesis by stimulating fibroblasts and cancer cells to produce pro-angiogenic proteases. ET 1 is a novel stimulator of tumour angiogenesis and warrants further examination as an anti-angiogenic treatment target. PMID- 16248774 TI - The pathobiology of endothelin-1 in vein graft disease: are ETA receptor antagonists the solution to prevent vein graft failure? AB - Despite the exploration of a large number of disparate drugs in animal models and clinical trials, no pharmacological intervention, with the exception of aggressive lipid lowering therapy has reduced late vein graft failure in man. The importance of devising more effective strategies is exemplified by the considerable economic consequences of vein graft failure. Worldwide, there are currently more than 1,000,000 coronary artery bypass graft surgery (CABG) operations a year, the same number of patients undergoing infrainguinal bypass (IIBS) for vascular diseases of the lower limb. The pathophysiology of vein graft failure is complex, involving disparate factors that include adhesion of platelets and leukocytes, rheological forces, metalloproteinase expression, proliferation and migration of vascular smooth muscle cells, neointima formation, oxidative stress, hypoxia and neural re-organisation. Although this diverse aetiology may seem to preclude any single drug type as being effective in preventing vein graft failure, one factor that is involved in every facet of vein graft pathobiology is endothelin-1 (ET-1). Thus, in this review, we will consider the diverse aetiology of vein graft disease in relation to ET-1 and will then present an argument (with evidence) that ET-1(A) (ET(A)) receptor antagonists constitute a potentially effective means of preventing vein graft failure. PMID- 16248775 TI - A role for endothelin-1 in peripheral vascular disease. AB - Peripheral vascular disease can compromise the blood supply to the lower limb with amputation being necessary in severe cases. Reduced blood flow may be due to arterial occlusive disease or constriction of skeletal microvessels with the resultant ischaemia causing pain, tissue damage, ulceration and gangrene. These events are associated with endothelial damage or dysfunction: endothelin-1 is implicated as a mediator via its constrictor, proinflammatory and proliferative actions. Raised plasma and tissue levels of this peptide have been described in various ischaemic conditions, including peripheral vascular disease. Here, the possible role of endothelin-1 in peripheral vascular disease is discussed and potential therapeutic tools are considered. PMID- 16248776 TI - Endothelin and the ischaemic heart. AB - Soon after its identification as a powerful vasoconstrictor peptide, endothelin (ET-1) was implicated as a detrimental agent involved in determining the outcome of myocardial ischaemia and reperfusion. Early experimental studies demonstrated that ET(A) selective and mixed ET(A)/ET(B) receptor antagonists can reduce infarct size and prevent ischaemia-induced ventricular arrhythmias in models of ischaemia/reperfusion, implying that ET-1 acts through the ET(A) receptor to contribute to injury and arrhythmogenesis. However, as our understanding of the physiology of ET-1 has expanded, the role of ET-1 in the ischaemic heart appears ever more complex. Recent evidence suggests that ET-1 exerts actions on the heart that are not only detrimental (vasoconstriction, inhibition of NO production, activation of inflammatory cells), but which may also contribute to tissue repair, such as inhibition of cardiomyocyte apoptosis. In addition, ET-1-induced mast cell degranulation has been linked to a homeostatic mechanism that controls endogenous ET-1 levels, which may have important implications for the ischaemic heart. Furthermore the mechanism by which ET-1 promotes arrhythmogenesis remains controversial. Some studies imply a direct electrophysiological effect of ET-1, via ET(A) receptors, to increase monophasic action potential duration (MAPD) and induce early after-depolarisations (EADs), while other studies support the view that coronary constriction resulting in ischaemia is the basis for the generation of arrhythmias. Moreover, ET-1 can induce cardioprotection (precondition) against infarct size and ventricular arrhythmias, through as yet incompletely understood mechanisms. To enable us to identify the most appropriate means of targeting this system in a therapeutically meaningful way we need to continue to explore the physiology of ET-1, both in the normal and the ischaemic heart. PMID- 16248777 TI - Endothelin signalling in the cardiac myocyte and its pathophysiological relevance. AB - Endothelin A (ET(A)) transmembrane receptors predominate in rat cardiac myocytes. These are G protein-coupled receptors whose actions are mediated by the G(q) heterotrimeric G proteins. Through these, ET-1 binding to ET(A)-receptors stimulates the hydrolysis of membrane phosphatidylinositol 4,5-bisphosphate to diacylglycerol and inositol 1,4,5-trisphosphate. Diacylglycerol remains in the membrane whereas inositol 1,4,5-trisphosphate is soluble (though its importance in the cardiac myocyte is still debated). Isoforms of the phospholipid-dependent protein kinase, protein kinase C (PKC), are intracellular receptors for diacylglycerol. Cytoplasmic nPKCdelta and nPKCepsilon detect increases in membrane diacylglycerols and translocate to the membrane. This brings about PKC activation, though modifications additional to binding to phospholipids and diacylglycerol are involved. The next event (probably associated with PKC activation) is the activation of the membrane-bound small G protein Ras by exchange of GTP for GDP. Ras.GTP loading translocates Raf family mitogen activated protein kinase (MAPK) kinase kinases to the membrane, initiates the activation of Raf, and thus activates the extracellular signal-regulated kinase 1/2 (ERK1/2) cascade. Over longer times, two analogous protein kinase cascades, the c-Jun N-terminal kinase and p38-mitogen-activated protein kinase cascades, become activated. As the signals originating from the ET(A) receptor are transmitted through these protein kinase pathways, other signalling molecules become phosphorylated, thus changing their biological activities. For example, ET 1 increases the expression of the c-jun transcription factor gene, and increases abundance and phosphorylation of c-Jun protein. These changes in c-Jun expression and phosphorylation are likely to be important in the regulation of gene transcription. PMID- 16248778 TI - Endothelin-1 and the aortic valve. AB - The aortic valve is a complex structure, the function of which is fundamental to sustain life. Previously believed to be an inert structure that merely opens in response to the forward flow of blood out of the left ventricle, it is now established that it is a sophisticated structure with specific biological properties. However, little is known about the mechanisms that regulate its function. In this respect, endothelin is of particular interest due to its range of biological actions within the cardiovascular system that suggest it may be capable of stimulating the cells that reside in valve cusps. Endothelin can be detected in the endothelial cells that cover valve cusps and it has been demonstrated that it is has the ability to stimulate contractile responses of cusp tissue in vitro. These contractions vary with different regions of the aortic valve cusp and occur preferentially in the circumferential direction. In addition, evidence exists that suggests endothelin may also have a role in the morphogenesis of the aortic valve. Further studies are required to determine the significance of the effects mediated by endothelin on cusp tissue to the function of the aortic valve in health and disease. PMID- 16248779 TI - Insulin resistance, obesity and the metabolic syndrome. Is there a therapeutic role for endothelin-1 antagonists? AB - There is increasing evidence to suggest that chronic activation of the endothelin 1 system can lead to heterologous desensitization of the glucose-regulatory and mitogenic actions of insulin with subsequent development of glucose intolerance, hyperinsulinemia, impaired endothelial function and exacerbation of cardiovascular disease. Effects are mediated through a variety of mechanisms that include attenuation of key insulin signalling pathways and decreased tyrosine phosphorylation of insulin receptor substrates IRS-1, SHC and G alpha q/11. Other actions involve hemodynamic changes leading to reduced delivery of insulin and glucose to peripheral tissues as well as enhanced hepatic glycogenolysis, decreased glucose-transporter translocation and modulation of various adipokines that regulate insulin action. Overall the data suggest that ET-1 antagonists may provide an effective means of improving cardiac dysfunction and favourably influencing glucose tolerance in obese humans and patients with early insulin sensitivity where there is clear evidence for activation of the ET-1 system. Although most effects of ET-1 that modulate mechanisms leading to glucose intolerance appear to involve the ETA receptor subtype recent data indicates that combined ETA/ETB receptor antagonists may function as effectively as selective ETA blockers. Prospective trials are needed to assess whether ET-1 antagonists, either alone or in combination, are superior to other more conventional therapies such as insulin sensitizers and to evaluate effects of combined treatments on the development of insulin resistance and the progression of diabetes. Early screening of patients at risk for evidence of ET-1 activation would help to identify subjects who may benefit most from such treatment. PMID- 16248780 TI - Endothelin and oxidative stress in the vascular system. AB - Both endothelin(ET)-1 and oxidative stress have been the subjects of intense investigation within the cardiovascular field over the past decade and a half, yet little is known about the precise relationship between these important modulators of vascular function. There is a firm evidence that ET-1 can stimulate the production of superoxide via NADPH oxidase activation, and at the same time, reactive oxygen species appear to stimulate ET-1 production. What is less clear is how these changes participate in the pathogenesis of vascular dysfunction. There is mixed evidence on whether oxidative stress plays a role in ET-dependent hypertension, however, a specific influence of ET-induced oxidative stress to reduce vascular reactivity is more convincing. The current review summarizes recent investigations into the relationship between ET-1 and oxidative stress and highlights several areas that require further investigation. PMID- 16248781 TI - Connective tissue remodeling: cross-talk between endothelins and matrix metalloproteinases. AB - Connective tissue remodeling is achieved by a complex process involving several cell types, a plethora of growth factors, cytokines, chemokines and turnover of extracellular matrix (ECM). The main enzymes that degrade ECM molecules are matrix metalloproteinases (MMPs) and their activities are regulated by endogenous inhibitors, the tissue inhibitors of metalloproteinases (TIMPs). Recent studies have indicated that endothelins and their receptor expression affects tissue remodeling and repair. Endothelins are rapidly produced by endothelial cells in response to tissue injury and they have potent vasoconstrictive properties. They also promote tissue remodeling through activation of resident connective tissue cells and controlling the production of MMPs and TIMPs by the activated cells. In this review we present the cross-talk between the endothelins and the MMP-TIMP system and their implications in controlling the normal and abnormal tissue remodeling. PMID- 16248782 TI - Localisation of endothelin-1 and its receptors in vascular tissue as seen at the electron microscopic level. AB - Since the discovery of endothelin in vascular endothelial cells and its pivotal role in vascular physiology (Yanagisawa and colleagues), a number of studies have focused on the localisation of this vasoconstrictor peptide in human and animal vascular tissue, largely in endothelial cells. Various vascular beds have been the subject of research in normal and pathophysiological conditions, for example in neonates, during ageing, pregnancy, hypertension, diabetes, heart failure, experimental metastases and neurological disorders. These studies have revealed the presence of endothelin in the blood vessel wall, suggesting the involvement of this peptide in vascular physiology in health and disease. This chapter reviews studies on the distribution of endothelin-1 (ET-1) and its receptors (ET(A) and ET(B)) in vascular tissue with emphasis on their ultrastructural localisation. PMID- 16248783 TI - Endothelin-1 and human platelets. AB - There is conflicting evidence regarding the effect of endothelin-1 (ET-1) on platelets. Some studies show that ET-1 activates platelets, others show platelet inhibition with ET-1 and some studies did not detect an effect of ET-1. These conflicting results may be due to complex interactions between platelet ET(A) and ET(B) receptors. ET-1 antagonism may emerge as an important therapeutic strategy in the management of several vascular disorders. However, to date the only prescribed ET-1 antagonist is bosentan for pulmonary arterial hypertension. Bosentan is a 'dual' ET-1 antagonist (i.e. it acts on both ET(A) and ET(B) receptors). Whether this action involves an effect on platelets remains to be established. In this review some of the studies describing the effect of ET-1 on human platelets are discussed. Vascular diseases where ET-1 is implicated are also considered. PMID- 16248785 TI - Design of Inhibitors for S100B. AB - S100B interacts with the p53 protein in a calcium-dependent manner and down regulates its function as a tumor suppressor. Therefore, inhibiting the S100B-p53 interaction represents a new approach for restoring functional wild-type p53 in cancers with elevated S100B such as found in malignant melanoma. A discussion of the biological rational for targeting S100B and a description of methodologies relevant to the discovery of compounds that inhibit S100B-p53 binding, including computational techniques, structural biology techniques, and cellular assays, is presented. PMID- 16248786 TI - Cell cycle regulatory kinase modulators: interim progress and issues. AB - Since a prior review of cell cycle inhibitors developed at the National Cancer Institute, continued progress in the application of these molecules has been pursued. Evidence of preliminary activity on the part of flavopiridol in certain chronic leukemias has pointed to that disease area as of potential interest, but likely by affecting transcriptional regulation through non-cell cycle-related CDKs. Brief duration infusion early phase trials with UCN-01, and combination studies with cytotoxics are commencing. Emerging structural data has refined the basis for screening strategies directed at cell cycle regulatory kinases, including cdks, chk kinases and most recently the mitotic phase aurora kinases. This interval progress report will review and update progress in these related but distinct drug discovery and development interest areas. PMID- 16248787 TI - Peptide nucleic acid conjugates: synthesis, properties and applications. AB - Artificial control of gene expression has great potential in the treatment of many human diseases, and peptide nucleic acids (PNAs) offer several potential advantages for silencing gene expression in mammalian cells. The pseudopeptide backbone of the PNA makes it resistant to enzymatic degradation, and PNAs bind complementary DNA and RNA with high affinity and specificity. PNAs are potentially leading agents for antigene and antisense therapeutics, but the application of PNAs in the in vivo setting is hampered by their poor intracellular delivery. This problem has been addressed by PNA conjugation to lipophilic moieties, peptides, and cell-specific receptor ligands. The biological activity of PNAs can also benefit from conjugation to DNA interactive compounds like intercalators and alkylators. Here we review the most interesting literature concerning PNA conjugation with small molecules, emphasizing synthetic approaches, properties and applications of the PNA conjugates. PMID- 16248788 TI - DNA mismatch repair deficiency, resistance to cancer chemotherapy and the development of hypersensitive agents. AB - DNA Mismatch Repair (MMR) deficiency results in resistance to platinating and alkylating agents, DNA minor groove binders, inhibitors of topoisomerases and antimetabolites. The cellular MMR pathway, involving hMLH1 and MSH2, detects and repairs DNA frame shifts replication errors and regulates recombination events. Tumour cells are able to cope with DNA damage caused by chemotherapy as long as the MMR-process is disabled and hence there is a need to develop agents that (i) restore MMR proficiency or (ii) are hypersensitive in cells that are irreversibly MMR deficient. Decitabine is suggested to restore MMR function by reversal of gene promoter hypermethylation of hMLH1. However, when MMR is deficient due to gene mutation it is not feasible to design agents, since the absence of functional proteins that constitute the MMR machinery are not available as targets. The evidence that resistance to chemotherapy is associated with hMSH2 and/or hMLH1 deficiency has revealed a new paradigm for drug discovery of agents that positively exploit this phenotype to therapeutic advantage. Even more attractive is the development of agents that are hypersensitive in the absence of functional MMR to enable even more effective treatment. In this regard, established agents such as mitomycin C, camptothecin or novel hydroxyethylaminoanthraquinones may represent opportunities for exploitation of MMR-deficiency in tumour cells. PMID- 16248790 TI - Carbohydrate mimotopes in the rational design of cancer vaccines. AB - The task of rationally designing vaccines that can effectively impact on the survival of cancer patients remains challenging. Monoclonal antibodies and T cell receptors have proven to be viable templates for the application of pharmacophore design principles to develop antigens and immunogens as these immune system molecules recognize a variety of sequentially and structurally unrelated ligands. This structural information combined with immunological assessment has contributed to the development of strategies to elicit effective humoral and cellular responses to cancer cells. Understanding the structural requirements for antibody and T cell recognition provides a basis for identifying potentially new sets of immunogens that may have both fundamental immunological and clinical value. Here we review the structural concepts and approaches used in vaccine design applications that illustrate the value and limitations of using chemical (peptide libraries) and immunological information to define novel peptide immunogens that function as mimotopes to generate immune responses targeting tumor associated carbohydrate antigens. PMID- 16248792 TI - NMR studies on how the binding complex of polyisoprenol recognition sequence peptides and polyisoprenols can modulate membrane structure. AB - The glycosyl carrier lipids, dolichylphosphate (C(95)-P) and undecapreylphosphate (C(55)-P) are key molecular players in the synthesis and translocation of complex glycoconjugates across cell membranes. The molecular mechanism of how these processes occur remains a mystery. Failure to completely catalyze C(95)-P mediated N-linked protein glycosylation is lethal, as are defects in the C(55)-P mediated synthesis of bacterial cell surface polymers. Our recent NMR studies have sought to understand the role these "super-lipids" play in biosynthetic and translocation pathways, which are of critical importance to problems in human biology and molecular medicine. The PIs can alter membrane structure by inducing in the lamellar phospholipids (PL) bilayer a non-lamellar or hexagonal (Hex(II)) structure. Membrane proteins that bind PIs contain a transmembrane binding motif, designated a PI recognition sequence (PIRS). Herein we review our recent combination of (1)H- and (31)P NMR spectroscopy and energy minimized molecular modeling studies that have determined the preferred orientation of PIs in model phospholipids membranes. They also show that the addition of a PIRS peptide to nonlamellar membranes induced by the PIs can reverse the Hex(II) phase back to a lamellar structure. Our molecular modeling calculations have also shown that as many as five PIRS peptides can bind to a single PI molecule. These findings lead to the hypothesis that the PI-induced Hex(II) structure may have the potential of forming a membrane channel that could facilitate glycoconjugate translocation processes. This is an alternate hypothesis to the possible existence of hypothetical "flippases" to accomplish movement of hydrophilic sugar chains across hydrophobic membranes. PMID- 16248793 TI - HIV-1 gp120 V3 loop for structure-based drug design. AB - HIV-1 cell entry is mediated by sequential interactions of the envelope protein gp120 with the receptor CD4 and a coreceptor, usually CCR5 or CXCR4, depending on the individual virion. Considerable efforts on exploiting the HIV coreceptors as drug targets have led to the new class of coreceptor antagonists. While these antiretroviral drugs aim at preventing virus/coreceptor interaction by binding to host proteins, neutralizing antibodies directed against the coreceptor-binding sites on gp120 have attracted attention as possible vaccine candidates. However, both approaches are complicated by the multiple protective mechanisms of gp120 which allow for rapid escape from selective pressures exerted by drugs or antibodies. Thus, advances in rational drug and vaccine design rely heavily on improved insights into the relation between genotype and phenotype, the evolution of coreceptor usage, and, ultimately the structural biology of coreceptor usage and inhibition. The third variable (V3) loop of gp120, crucially involved in all these aspects, will be a major focus of this review. PMID- 16248794 TI - Progress in protein structural class prediction and its impact to bioinformatics and proteomics. AB - The structural class is an important attribute used to characterize the overall folding type of a protein or its domain. Since the concept of protein structural class was developed about 3 decades ago based on a visual inspection of polypeptide chain topologies in a dataset of only 31 gloular proteins, the number of structure-known proteins has been increased rapidly. For example, as of 12 July-2005, the entries deposited into RCSB PDB Protein Data Bank for proteins, peptides, and viruses whose 3-dimensional structures were determined by X-ray and NMR techniques have been increased to 28,920. To properly cover more and more structure-known proteins, some modification and expansion from the original structural classification scheme have been developed. Meanwhile, many different approaches have been proposed for predicting the structural class of proteins. In this review, the new classification schemes are briefly introduced. The attention is focused on the progress in structural class prediction and its impact in stimulating the development of identifying the other attributes of proteins. It is interesting to point out that the development of the latter has actually in turn greatly enriched the power of the former. Also, some promising approaches for the further development of protein structural class prediction are also addressed. PMID- 16248795 TI - Analysis of the phosphoryl transfer mechanism of c-AMP dependent protein kinase (PKA) by penta-coodinate phosphoric transition state theory. AB - This review briefly covers recent literature of research on the phosphoryl transfer mechanism of PKA. Combining experimental and theoretical calculation results on enzymes with experimentally observed biomimic activities of phosphoryl amino acids and a small molecular model of catalytic core in PKA, a novel mechanism was proposed. The cooperative participation roles of both Asp166 and Lys168 via a penta-coodinate phosphoric intermediate was elucidated to conciliate the current different views of the phosphoryl transfer mechanism of PKA. Since many ATP-binding enzymes may share a similar phosphoryl transfer mechanism, this proposed mechanism might also apply to the mechanism of these enzymes, e.g., molecular motor and phosphatase among others. PMID- 16248797 TI - Anesthetics as chemical tools to study the structure and function of nicotinic acetylcholine receptors. AB - The nicotinic acetylcholine receptor (AChR) is the archetype of the Cys-loop ligand-gated ion channel receptor superfamily. Noncompetitive antagonists inhibit the AChR without interacting directly with agonist sites. Among noncompetitive antagonists, general and local anesthetics have been used for decades to study the structure and function of muscle- as well as neuronal-type AChRs. In this review, we address and update all information regarding the characterization of binding sites and the mechanism of action for n-alkanols, barbiturates, inhalational and dissociative general anesthetics, as well as for tertiary and quaternary local anesthetics. The experimental evidence outlined in this review suggest that: (1) several neuronal-type AChRs might be targets for the pharmacological action of distinct anesthetics; (2) the molecular components of a specific anesthetic locus on a certain receptor type are different from the structural determinants of the site for the same anesthetic on a different receptor type; (3) there are unique binding sites for distinct anesthetics in the same receptor; (4) the affinity of a specific anesthetic depends on the AChR conformational state; (5) anesthetics may inhibit AChRs by different mechanisms including open-channel-blocking, augmenting the desensitization process, and/or inactivating the opening of resting receptors; and (6) some anesthetics may potentiate AChR activity. PMID- 16248796 TI - Computational methods for protein-protein interaction and their application. AB - Protein-protein interactions play a central role in numerous processes in cell and are one of the main research fields in current functional proteomics. The increase of finished genomic sequences has greatly stimulated the progress for detecting the functions of the genes and their encoded proteins. As complementary ways to the high through-put experimental methods, various methods of bioinformatics have been developed for the study of the protein-protein interaction. These methods range from the sequence homology-based to the genomic context based. Recently, it tends to integrate the data from different methods to build the protein-protein interaction network, and to predict the protein function from the analysis of the network structure. Efforts are ongoing to improve these methods and to search for novel aspects in genomes that could be exploited for function prediction. This review highlights the recent advances of the bioinformatics methods in protein-protein interaction researches. In the end, the application of the protein-protein interaction has also been discussed. PMID- 16248798 TI - Conserved molecular players for axon guidance and angiogenesis. AB - Neuronal guidance cues attract or repel axons and/or neurons and play important roles in the pathfinding of neuronal networks and the functioning of nervous system. Prominent among them are the families of ephrins, semaphorins, Slits and netrins and their cognate cell-surface receptors. Due to their biological significance, extensive research has been carried out in the last ten years or so. Angiogenesis is a cellular process of capillary sprouting and configuring of neovasculatures, which shares many developmental, anatomical, physiological and pathophysiological features with the neural counterparts. This review will summarize the emerging evidence indicating the common molecular mechanisms underlying both axon guidance (including neuronal migration) and angiogenesis for exquisite regulation of proper wiring of both systems. PMID- 16248789 TI - Combining cytotoxic and immune-mediated gene therapy to treat brain tumors. AB - Glioblastoma (GBM) is a type of intracranial brain tumor, for which there is no cure. In spite of advances in surgery, chemotherapy and radiotherapy, patients die within a year of diagnosis. Therefore, there is a critical need to develop novel therapeutic approaches for this disease. Gene therapy, which is the use of genes or other nucleic acids as drugs, is a powerful new treatment strategy which can be developed to treat GBM. Several treatment modalities are amenable for gene therapy implementation, e.g. conditional cytotoxic approaches, targeted delivery of toxins into the tumor mass, immune stimulatory strategies, and these will all be the focus of this review. Both conditional cytotoxicity and targeted toxin mediated tumor death, are aimed at eliminating an established tumor mass and preventing further growth. Tumors employ several defensive strategies that suppress and inhibit anti-tumor immune responses. A better understanding of the mechanisms involved in eliciting anti-tumor immune responses has identified promising targets for immunotherapy. Immunotherapy is designed to aid the immune system to recognize and destroy tumor cells in order to eliminate the tumor burden. Also, immune-therapeutic strategies have the added advantage that an activated immune system has the capability of recognizing tumor cells at distant sites from the primary tumor, therefore targeting metastasis distant from the primary tumor locale. Pre-clinical models and clinical trials have demonstrated that in spite of their location within the central nervous system (CNS), a tissue described as 'immune privileged', brain tumors can be effectively targeted by the activated immune system following various immunotherapeutic strategies. This review will highlight recent advances in brain tumor immunotherapy, with particular emphasis on advances made using gene therapy strategies, as well as reviewing other novel therapies that can be used in combination with immunotherapy. Another important aspect of implementing gene therapy in the clinical arena is to be able to image the targeting of the therapeutics to the tumors, treatment effectiveness and progression of disease. We have therefore reviewed the most exciting non-invasive, in vivo imaging techniques which can be used in combination with gene therapy to monitor therapeutic efficacy over time. PMID- 16248799 TI - Pattern recognition methods for protein functional site prediction. AB - Protein functional site prediction is closely related to drug design, hence to public health. In order to save the cost and the time spent on identifying the functional sites in sequenced proteins in biology laboratory, computer programs have been widely used for decades. Many of them are implemented using the state of-the-art pattern recognition algorithms, including decision trees, neural networks and support vector machines. Although the success of this effort has been obvious, advanced and new algorithms are still under development for addressing some difficult issues. This review will go through the major stages in developing pattern recognition algorithms for protein functional site prediction and outline the future research directions in this important area. PMID- 16248800 TI - Cancer vaccine and immunotherapy. PMID- 16248801 TI - Melanoma immunotherapy: past, present, and future. AB - The incidence of cancer and its related morbidity and mortality remain on the increase in both developing and developed countries. Cancer remains a huge burden on the health and social welfare sectors worldwide and its prevention and cure remain two golden goals that science strives to achieve. Among the treatment options for cancer that have emerged in the past 100 years, cancer vaccine immunotherapy seems to present a promising and relatively safer approach as compared to chemotherapy and radiotherapy. The identification of different tumour antigens in the last fifteen years using a variety of techniques, together with the molecular cloning of cytotoxic T lymphocytes (CTLs)- and tumour infiltrating lymphocytes (TILs)-defined tumour antigens allowed more refining of the cancer vaccines that are currently used in different clinical trials. In a proportion of treated patients, some of these vaccines have resulted in partial or complete tumour regression, while they have increased the disease-free survival rate in others. These outcomes are more evident now in patients suffering from melanoma. This review provides an update on melanoma vaccine immunotherapy. Different cancer antigens are reviewed with a detailed description of the melanoma antigens discovered so far. The review also summarises clinical trials and individual clinical cases in which some of the old and current methods to vaccinate against or treat melanoma were used. These include vaccines made of autologous or allogenic melanoma tumour cells, melanoma peptides, recombinant bacterial or viral vectors, or dendritic cells. PMID- 16248802 TI - Immunotherapy and cancer vaccines in the management of breast cancer. AB - Besides the traditional therapeutic options, treatment with antibodies specific for the receptor tyrosine kinase HER-2/neu has been established as a standard therapy in the clinical management of advanced breast cancer. Ongoing clinical studies focus on the improvement of application protocols in order to minimize side effects and evaluate the potential therapeutic benefit of anti-HER-2/neu antibodies in combination with conventional chemotherapy. Various similar strategies to target other tumour-associated antigens or proangiogenic factors with inhibitory antibodies are currently investigated in promising preclinical and clinical trials. In addition, research efforts are made to develop procedures to generate tumour-specific cellular immune responses in breast cancer patients. Therapeutic vaccination is, however, still at an early stage of development, despite encouraging results of animal studies. We summarise and discuss vaccination strategies with tumour-specific proteins or peptides, pulsed dendritic cells, and modified tumour cells as well as antibody-based therapeutic concepts to target HER-2/neu, EGF receptor, MUC-1, uPA/uPAR, and VEGF. PMID- 16248803 TI - Therapeutic vaccines for cervical cancer: dendritic cell-based immunotherapy. AB - Human papillomavirus (HPV) infection represents the most important risk factor for the development of cervical dysplasia and cervical cancer. Several lines of evidence suggest that cell-mediated immune responses are important in controlling both HPV infections and HPV-associated neoplasia. Since HPV E6 and E7 oncoproteins are expressed in these lesions and are necessary for the maintenance of the malignant phenotype, these proteins might be potential tumor-specific target antigens for immunotherapy of cervical cancer. The gold standard treatment for locally advanced cervical cancer is primary radiation therapy combined with chemotherapy. A potential drawback of this potentially curative treatment is a profound and long lasting negative effect on the immune system. Treatment-induced immunosuppression combined with tumor-induced subversion of the immune system may therefore impose severe limitations on the efficacy of conventional vaccination strategies in late stage cervical cancer patients. The recognition of dendritic cells (DC) as powerful antigen-presenting cells capable of inducing primary T cell responses in vitro and in vivo, has recently generated widespread interest in DC-based immunotherapy of several human malignancies. Here, we review various therapeutic HPV vaccines being developed and implemented in human clinical trials, with a particular emphasis on the use of autologous DC pulsed with full length HPV 16 or 18 E7 oncoproteins as a novel strategy to induce HPV E7-specific and tumor-specific T cell responses in cervical cancer patients following conventional treatment. PMID- 16248804 TI - Immunity to tumour antigens. AB - During the last decade, a large number of human tumour antigens have been identified. These antigens are classified as tumour-specific shared antigens, tissue-specific differentiation antigens, overexpressed antigens, tumour antigens resulting from mutations, viral antigens and fusion proteins. Antigens recognised by effectors of immune system are potential targets for antigen-specific cancer immunotherapy. However, most tumour antigens are self-proteins and are generally of low immunogenicity and the immune response elicited towards these tumour antigens is not always effective. Strategies to induce and enhance the tumour antigen-specific response are needed. This review will summarise the approaches to discovery of tumour antigens, the current status of tumour antigens, and their potential application to cancer treatment. PMID- 16248805 TI - Use of cytokines in cancer vaccines/immunotherapy: recent developments improve survival rates for patients with metastatic malignancy. AB - Historically, patients with disseminated cancer have had poor prognoses and chemotherapy has been of little benefit. However, several different avenues of clinical research are providing reasons for hope. The advent of cytokine immunotherapy, particularly in combination with chemotherapy (biochemotherapy) has seen significantly improved outcomes for metastatic disease. Early biochemotherapy trials often revealed more than 20% complete responses. Unfortunately, Phase III trials have not confirmed earlier expectations for reasons that are not clear, but may reflect the inclusion of patients with refractory brain or other metastases in later trials. More recently, cancer vaccine therapies have provided significantly improved patient survival rates. It is not uncommon for 5-year survival rates of post-surgical patients recovering from metastatic malignancy who receive cancer vaccine therapy to reach more than 50%. Cytokines have become an integral part of cancer therapy and are also under trial together with cancer vaccines as post-surgical adjuvant therapies providing significant gains in long term survival rates. New insights from several different areas of research into the properties of tumour cells and their significance for immunosurveillance point to the importance of the tumour cells themselves as antigen presenting cells. Recent developments with genetically deficient animals and cancer cells have provided greater understanding at the molecular level of the importance of a functioning antigen presenting system operating inside tumour cells. This new knowledge offers support for further enhancing patient survival by combining previous therapies such as use of cytokines in biochemotherapy together with immunization using cytokine activated whole cell cancer vaccines in the future. PMID- 16248806 TI - Direct evidence on the immune-mediated spontaneous regression of human cancer: an incentive for pharmaceutical companies to develop a novel anti-cancer vaccine. AB - To develop an effective pharmaceutical treatment for a disease, we need to fully understand the biological behavior of that disease, especially when dealing with cancer. The current available treatment for cancer may help in lessening the burden of the disease or, on certain occasions, in increasing the survival of the patient. However, a total eradication of cancer remains the researchers' hope. Some of the discoveries in the field of medicine relied on observations of natural events. Among these events is the spontaneous regression of cancer. It has been argued that such regression could be immunologically-mediated, but no direct evidence has been shown to support such an argument. We, hereby, provide compelling evidence that spontaneous cancer regression in humans is immunologically-mediated, hoping that the results from this study would stimulate the pharmaceutical industry to focus more on cancer vaccine immunotherapy. Our results showed that patients with >3 primary melanomas (very rare group among cancer patients) develop significant histopathological spontaneous regression of further melanomas that they could acquire during their life (P=0.0080) as compared to patients with single primary melanoma where the phenomenon of spontaneous regression is absent or minimal. It seems that such regression resulted from the repeated exposure to the tumor which mimics a self-immunization process. Analysis of the regressing tumors revealed heavy infiltration by T lymphocytes as compared to non-regressing tumors (P<0.0001), the predominant of which were T cytotoxic rather than T helper. Mature dendritic cells were also found in significant number (P<0.0001) in the regressing tumors as compared to the non regressing ones, which demonstrate an active involvement of the different arms of the immune system in the multiple primary melanoma patients in the process of tumor regression. Also, MHC expression was significantly higher in the regressing versus the non-regressing tumors (P <0.0001), which reflects a proper tumor antigen expression. Associated with tumor regression was also loss of the melanoma common tumor antigen Melan A/ MART-1 in the multiple primary melanoma patients as compared to the single primary ones (P=0.0041). Furthermore, loss of Melan A/ MART-1 in the regressing tumors significantly correlated with the presence of Melan A/ MART-1-specific CTLs in the peripheral blood of these patients (P=0.03), which adds to the evidence that the phenomenon of regression seen in these patients was immunologically-mediated and tumor-specific. Such correlation was also seen in another rare group of melanoma patients, namely those with occult primary melanoma. The lesson that we could learn from nature in this study is that inducing cancer regression using the different arms of the immune system is possible. Also, developing a novel cancer vaccine is not out of reach. PMID- 16248808 TI - Dopamine-glutamate interaction and antipsychotics mechanism of action: implication for new pharmacological strategies in psychosis. AB - Schizophrenia is a severe mental illness characterized by behavioral and cognitive symptoms. Several lines of evidence focus on a direct involvement of the glutamatergic system in the pathophysiology of psychosis. The hypofunction of the ionotropic glutamate N-methyl-D-Aspartate Receptor (NMDA-R) has been proposed as a model of schizophrenia in humans. Cortical and subcortical glutamate release seems to be modulated by dopaminergic and, to a lesser extent, serotoninergic circuitries, and tuned by intracellular pathways. Although dopamine D(2) receptor blockade is a crucial mechanism of antipsychotics pharmacodynamic profile, a putative glutamatergic impact of these compounds is suggested by animal pharmacological isomorphisms of psychosis as well as by clinical studies. According to this view, the balance between D(2) antagonism and NMDA-R modulation may be pivotal for the improvement of both positive and negative symptoms. Recently, many pharmacological strategies involving glutamate receptors have been suggested, and novel compounds and pharmacological strategies acting on glutamate transmission are currently under evaluation: i) augmentation strategies improving NMDA-R transmission (glycine, D-serine, D-cycloserine, glycine transporter inhibitors); ii) ampakines, positive modulators of AMPA (alpha-amino-3-hydroxy-5 methyl-4-isoxazole propionic acid) receptor complex; iii) agonists of glutamate metabotropic receptors; iv) drugs involved in subcellular adaptation both at pre- and post-synaptic sites. Furthermore, molecular markers, suggesting modulation of glutamate circuitries after antipsychotics administration, are an attractive tool to shed more light on glutamate involvement in antipsychotics mechanism of action. In this review we provide a critical update of recent preclinical and clinical data on dopamine-glutamate interaction and its role in new pharmacological strategies for psychosis treatment. PMID- 16248807 TI - Why drugs fail--a study on side effects in new chemical entities. AB - Over 90% of the market withdrawals were caused by drug toxicity. Hepatotoxicity and cardiovascular toxicity proved to be the major causes for two out of three market withdrawals in the respective time period. In clinical phases I-III 43% of drug development project terminations were due to insufficient efficacy of the investigated compound. The second important issue, which caused one third of the projects to be closed, was toxicity. ADME parameters and economic and other reasons played a minor role. The results of our study indicate that compared with previous studies on this subject, no major improvements have been achieved in the last decade. PMID- 16248809 TI - Comparison of different fluorescence fluctuation methods for their use in FRET assays: monitoring a protease reaction. AB - We compare the accuracy of a variety of Fluorescence Fluctuation Spectroscopy (FFS) methods for the study of Forster Resonance Energy Transfer (FRET) assays. As an example, the cleavage of a doubly labeled, FRET-active peptide substrate by the protease Trypsin is monitored and analyzed using methods based on fluorescence intensity, Fluorescence Correlation Spectroscopy (FCS) and Fluorescence Intensity Distribution Analysis (FIDA). The presented fluorescence data are compared to High-Pressure Liquid Chromatography (HPLC) data obtained from the same assay. The HPLC analysis discloses general disadvantages of the FRET approach, such as incomplete labeling and the need for aliquots. However, the simultaneous use of two photon detectors monitoring the fluorescence signal of both labels significantly improves the analysis. In particular, the two global analysis tools Two-Dimensional Fluorescence Intensity Distribution Analysis (2D FIDA) and Two-Color Global Fluorescence Correlation Spectroscopy (2CG-FCS) highlight the potential of a combination of FFS and FRET. While conventional FIDA and FCS auto- or cross-correlation analysis leaves the user with drawbacks inherent in two-color and FRET applications, these effects are overcome by the global analysis on the molecular level. Furthermore, it is advantageous to analyze the unnormalized as opposed to the normalized correlation data when combining any fluorescence correlation method with FRET, since the analysis of the unnormalized data introduces more accuracy and is less sensitive to the experimental drawbacks. PMID- 16248811 TI - Diffusion and binding properties investigated by Fluorescence Correlation Spectroscopy (FCS). AB - During the last years, Fluorescence Correlation Spectroscopy (FCS) has proven to be a powerful tool for basic research in many applications. The combination of a minimal detection volume in the femtoliter range coupled with very high sensitivity extends the possibilities to design sensitive homogeneous tests. In this article we illustrate the analysis of binding processes with FCS based on the changes in diffusion characteristics of GFP upon binding to an antibody. Problems induced by highly heterogeneous samples are discussed and differences of GFP binding to a monoclonal and a polyclonal antibody are shown and analyzed. We stress data processing, limitations and useful approximations in FCS methodology. Basic ideas of data acquisition and processing as well as new developments and applications are presented. PMID- 16248810 TI - Nutritional and therapeutic potential of Spirulina. AB - Spirulina, a filamentous cyanobacterium, possesses diverse biological activities and nutritional significance due to high concentration of natural nutrients, having bio-modulatory and immuno-modulatory functions. Different Spirulina preparations influence immune system viz. increase phagocytic activity of macrophages, stimulating the production of antibodies and cytokines, increase accumulation of NK cells into tissue and activation and mobilization of T and B cells. Spirulina have also shown to perform regulatory role on lipid and carbohydrate metabolism by exhibiting glucose and lipid profile correcting activity in experimental animals and in diabetic patients. Preparations have been found to be active against several enveloped viruses including herpes virus, cytomegalovirus, influenza virus and HIV. They are capable to inhibit carcinogenesis due to anti-oxidant properties that protect tissues and also reduce toxicity of liver, kidney and testes. PMID- 16248812 TI - Advanced approaches in insulin delivery. AB - Diabetes is a syndrome of disordered metabolism and inappropriate hyperglycemia resulting from a deficiency of insulin secretion or insulin resistance. Insulin, a pancreatic hormone, helps to lower the blood sugar levels. The structural features of insulin and insulin receptors are summarized. Diabetic patients use insulin in the form of injections, which involves lots of pain, and a need for non-invasive, alternative mode of insulin administration is desired. These challenges have lead to attempts in insulin therapy using oral, nasal, pulmonary, rectal, transdermal, buccal, gene therapy, islet cell transplantation and diabetes vaccine. Among all the approaches pulmonary administration has achieved some clinical significance. Future approaches that can be exploited for insulin therapy in Insulin Dependent Diabetes Mellitus [IDDM] have been summarized. Insulin inhalers or tablets for IDDM are interesting alternatives. PMID- 16248813 TI - The bioreactor: a powerful tool for large-scale culture of animal cells. AB - Bioreactors play a key role in the field of biologics, where they are used for the production of recombinant therapeutic proteins by large-scale cultivation of animal cells. There are several types of bioreactors, including stirred-tank, airlift, hollow-fiber, and Rotary Cell Culture System (RCCS) designs. The stirred tank bioreactor is one of the most commonly used types, and is used both for industrial applications and laboratory research. The RCCS, invented by NASA, is increasingly used in the area of tissue engineering for medical purposes. Important improvements have been made in the design of traditional bioreactors, and new types of bioreactor are also being developed such as Couette-Taylor bioreactor, multifunctional-membrane bioreactor, and shaking bioreactor. Work is also progressing on techniques to improve the performance of bioreactors, including perfusion culture, the use of microcarriers, and methods of suppressing apoptosis and of monitoring cell growth in real time. Given the demand for the production by animal cells for use in the growing number of clinical applications, further advances in bioreactor technology can be expected during the next few years. Two main goals will be pursued: firstly, to increase output by high density cultivation of animal cells to produce high value protein pharmaceutics or viral vectors for clinical gene therapy; and secondly, to create a three-dimension space similar to that of an in vivo environment to regenerate tissue or organ and to reproduce valuable cells that are hard to culture in the traditional culture system. PMID- 16248814 TI - Enhancing the sensitivity of fluorescence correlation spectroscopy by using time correlated single photon counting. AB - Fluorescence correlation spectroscopy (FCS) and fluorescence cross-correlation spectroscopy (FCCS) are methods that extract information about a sample from the influence of thermodynamic equilibrium fluctuations on the fluorescence intensity. This method allows dynamic information to be obtained from steady state equilibrium measurements and its popularity has dramatically increased in the last 10 years due to the development of high sensitivity detectors and its combination with confocal microscopy. Using time-correlated single-photon counting (TCSPC) detection and pulsed excitation, information over the duration of the excited state can be extracted and incorporated in the analysis. In this short review, we discuss new methodologies that have recently emerged which incorporated fluorescence lifetime information or TCSPC data in the FCS and FCCS analysis. Time-gated FCS discriminates between which photons are to be incorporated in the analysis dependent upon their arrival time after excitation. This allows for accurate FCS measurements in the presence of fluorescent background, determination of sample homogeneity, and the ability to distinguish between static and dynamic heterogeneities. A similar method, time-resolved FCS can be used to resolve the individual correlation functions from multiple fluorophores through the different fluorescence lifetimes. Pulsed interleaved excitation (PIE) encodes the excitation source into the TCSPC data. PIE can be used to perform dual-channel FCCS with a single detector and allows elimination of spectral cross-talk with dual-channel detection. For samples that undergo fluorescence resonance energy transfer (FRET), quantitative FCCS measurements can be performed in spite of the FRET and the static FRET efficiency can be determined. PMID- 16248815 TI - Confocal fluorescence microscopy for high-throughput screening of G-protein coupled receptors. AB - In the pharmaceutical industry, G-protein coupled receptors (GPCRs) are the most successful group of therapeutic targets. Finding compounds that interfere with the ligand-GPCR interaction in a specific and selective way is a major focus of pharmaceutical research today. As compound libraries of large pharmaceutical companies have increased to hundreds of thousands of test compounds, there is a growing need for miniaturization of drug discovery assays to save bioreagents and to reduce the consumption of test compounds. Due to its high sensitivity combined with a femtoliter-sized measurement volume, confocal fluorescence microscopy enables designs for GPCR binding assays with tiny sample volumes. The GPCRs are prepared in the form of plasma membrane fragments from GPCR-overexpressing cells or may be integrated into virus-like particles (VLiPs). One technique to extract binding data from confocal fluorescence experiments is the so-called fluorescence intensity distribution analysis (FIDA). In this review article, we describe the applicability of FIDA to GPCR-focussed high-throughput screening (HTS) and compare FIDA to two other GPCR-adaptable drug discovery techniques for ligand binding studies, the scintillation proximity assay (SPA) and macroscopic fluorescence polarization (FP) measurements. FIDA measures the absolute concentrations of both GPCR-bound and unbound ligand, thereby providing an internal control to the drug screening data. FIDA is amenable to work with relatively low amounts of GPCRs so that the assay may be carried out with biomembranes of a low GPCR density. Moreover, the fluorescence intensity readout of the FIDA technique may be combined with other confocal fluorescence readouts such as fluorescence anisotropy or lifetime. The combination of a low sample volume with an information-rich measurement means that confocal fluorescence spectroscopy can bring substantial benefits as a bioassay platform to pharmaceutical GPCR-directed research. PMID- 16248816 TI - Peptide nucleic acids (PNAs), a chemical overview. AB - Peptide nucleic acid (PNA) is a nucleic acid analogue and a fully synthetic DNA/RNA-recognising ligand with a neutral peptide-like backbone. In spite of the large change on the backbone structure, PNA molecules bind strongly to complementary DNA and RNA sequences. Originally conceived as ligand for the recognition of double stranded DNA, the unique physico-chemical properties of PNAs have led to the development of a variety of research and diagnostic assays. The extraordinary properties of PNA may advance routine clinical tests and environmental analyses that will utilise the PNA technology. PNAs may also have an impact on in situ hybridisation, cytogenetics and industrial microbiology. This paper presents some recent achievements on peptide nucleic acids and discusses, from the viewpoint of literature, what the potential is and what the limitations of such compounds are. This review, which is not intended to be exhaustive, is mostly aimed at the current progress in PNA chemistry, structure, and hybridisation, highlighting some applications too. PMID- 16248817 TI - Mechanism of mitochondrial uncouplers, inhibitors, and toxins: focus on electron transfer, free radicals, and structure-activity relationships. AB - The biology of the mitochondrial electron transport chain is summarized. Our approach to the mechanism of uncouplers, inhibitors, and toxins is based on electron transfer (ET) and reactive oxygen species (ROS). Extensive supporting evidence, which is broadly applicable, is cited. ROS can be generated either endogenously or exogenously. Generally, the reactive entities arise via redox cycling by ET functionalities, such as, quinones (or precursors), metal compounds, imines (or iminiums), and aromatic nitro compounds (or reduced metabolites). In most cases, the ET functions are formed metabolically. The toxic substances belong to many categories, e.g., medicinals, industrial chemicals, abused drugs, and pesticides. Structure-activity relationships are presented from the ET-ROS perspective, and also quantitatively. Evidence for the theoretical framework is provided by the protective effect of antioxidants. Among other topics addressed are proton flux, membrane pores, and apoptosis. There is support for the thesis that mitochondrial insult may contribute to illnesses and aging. PMID- 16248818 TI - Medicinal plants with inhibitory properties against snake venoms. AB - Envenomations due to snake bites are commonly treated by parenteral administration of horse or sheep-derived polyclonal antivenoms aimed at the neutralization of toxins. However, despite the widespread success of this therapy, it is still important to search for different venom inhibitors, either synthetic or natural, that could complement or substitute for the action of antivenoms. Several plants have been utilized in folk medicine as antiophidian. However, only a few species have been scientifically investigated and still less had their active components isolated and characterized both structurally and functionally. This article presents a review of plants showing neutralizing properties against snake venoms which were assayed in research laboratories, correlating them with ethnopharmacological studies, as (i) the part of the plant used as antidote, (ii) its respective genus and family and (iii) inhibition of the main pharmacological, toxic and enzymatic activities of snake venoms and isolated toxins. Protective activity of many of these plants against the lethal action of snake venoms has been confirmed by biological assays. Compounds in all of them belong to chemical classes capable of interacting with macromolecular targets (enzymes or receptors). Popular culture can often help to guide scientific studies. In addition, biotechnological application of these inhibitors, as helpful alternative or supplemental treatments to serum therapy, and also as important models for synthesis of new drugs of medical interest, needs to be better oriented and scientifically explored. PMID- 16248819 TI - Current status of malaria control. AB - Malaria caused by Plasmodium parasites kills approximately 1-3 million people and causes disease in 300-500 million people annually throughout the world. The current approaches to curtail this disease include vector control, vaccination, immunotherapy and chemotherapy. The vector control is achieved by reducing vector density, interrupting their life cycle, and creating a barrier between the human host and mosquitoes. A number of vaccine candidates are being clinically tried and R&D effort in this direction is coming in a big way. Currently there are only limited safe drugs for the treatment of this disease, however, reports of emerging resistance against existing drugs warrant the introduction of new drugs, which are unlikely to come from pharmaceutical industries because of limited commercial opportunities. One of the most important current approaches to develop new drugs involves the synthesis of chemical libraries and evaluate them against most validated biochemical targets of malarial parasite. Although a number of such targets in antimalarial drug development are known today, yet only validated and selective biochemical targets including mitochondrial transport, glycolic pathway, folate pathway, proteases and heme metabolism, apicoplast metabolism, glycophospatidyl inositol, lipid metabolism (glycerophospholipids), peptidyl deformylase and oxidative stress in parasite-infected erythrocytes have been discussed here. The well known antimalarial drugs and different drug combinations for the treatment of malaria are also briefly reviewed. A survey of the recently discovered new molecules active against malaria has also been narrated. Lastly, the future of malaria chemotherapy and new directions emerging from literature has been elucidated. PMID- 16248821 TI - Perinatal events affecting the onset of allergic diseases. AB - The prevalence of asthma and related allergic disorders has increased considerably over the last several decades. Since the genetic makeup of humans has not changed during this time, it is likely that environmental factors may have influenced this rise in allergic diseases. Furthermore, there is increasing evidence to suggest that many aspects of health and disease are determined during the perinatal period and that alterations in lifestyle and diet later in life are secondary to the effects of the immunological programming that occurs during pregnancy and early infancy. This is directly applicable to allergic disease where immune responses at birth implicate intrauterine exposure as a primary sensitization event. Moreover, infants who experience allergy early in life already have an altered immune response at birth and most therapeutic approaches focus on altering the expression of the disease. Therefore, a better understanding of the underlying mechanisms that shapes the immune response towards allergy development is fundamental to strengthening "natural" protective stimuli or developing preventative rather than treatment therapies. PMID- 16248822 TI - Skin barrier dysfunction and systemic sensitization to allergens through the skin. AB - Most allergic, atopic and hypersensitive reactions are associated with Th2-biased immune responses and allergen-specific IgE antibodies. Pathological allergic disorders are on an alarming increase in the industrialized world. Understanding the mechanism of primary sensitization to allergens is important in elucidating the pathogenesis of these diseases and for possibly preventing their development. In this article, we review recent information supporting that epidermal allergen exposure may contribute to systemic allergic diseases and that atopy may be secondary to skin barrier dysfunction in some dermatoses. The skin is an active immunological organ, which functions as a primary defence and biosensor to the external environment. The critical permeability barrier function is mediated by the outmost layer of the epidermis, the stratum corneum. Perturbation of the stratum corneum initiates a chain of event, which activates homeostatic responses in the underlying epidermis. Repeated barrier-disruption, whether environmentally or genetically determined, may however stimulate signaling cascades that lead to inflammation and epidermal hyperplasia. Skin barrier dysfunction may also allow entry of allergens, which can lead to primary systemic sensitization. The altered epidermal microenvironment in barrier-disrupted skin appears to be particularly well suited for the induction of potent Th2-type responses with production of allergen-specific IgE. Epidermal exposure to food antigens can prevent the normal induction of oral tolerance and also lead to airway eosinophilia following inhalation. Exposure to allergens on barrier-disrupted skin may as such serve as a natural sensitization pathway for food allergy and respiratory allergic disease. PMID- 16248823 TI - Pharmaceutical treatment of asthma in children. AB - The present review article gives an overview of the present treatment modalities of asthma during childhood and discusses the existing controversies in asthma treatment. Present guidelines of asthma treatment concentrate on treatment for adults and only marginally concern treatment of childhood asthma. The few exceptions are the British Scottish guidelines and the Nordic guidelines, which have separate paragraphs on paediatric asthma management. The main controversy in paediatric asthma treatment is that how early (in age) and how soon (after diagnosis of asthma has been established) should inhaled steroids be instituted. Does treatment with early inhaled steroids influence lung development? Also possible side effects of inhaled steroids as possible impact upon growth and effect upon the hypothalamic adrenal axis are discussed. What is the place of leukotriene antagonists in childhood asthma treatment? Other issues discussed are prevention of asthma (primary, secondary and tertiary) in relationship to treatment of asthma. Primary prevention regards preventive measures to be taken to prevent initial allergic diseases; secondary prevention aims at preventing development of further allergic disease after the initial allergic disorder, as preventing debut of asthma after atopic eczema. Tertiary prevention aims at reducing already existing allergic illness and preventing further progression. For asthma, tertiary prevention regards treatment. During later years, there has been a focus on the respiratory tract as a continuum, and how allergic rhinitis and asthma should be treated when they are coexistent. Treating exercise induced asthma optimally is regarded as an important aim in the general treatment of asthma in childhood. Particularly in childhood asthma, compliance (concordance) with treatment is an important issue. Also some controversial aspects of acute asthma treatment in young children are discussed. PMID- 16248824 TI - Anti-IgE therapy in allergic asthma. AB - Despite the advanced and increasing understanding of the pathophysiology of asthma and other allergic diseases, current treatment remains unspecific and targets late events within the allergic cascade. Therefore, a novel and promising approach to treat allergic asthma is antagonizing IgE by anti-IgE antibodies. This review analyzes the role of IgE for the pathology of asthma, summarize the current data about action, safety and efficacy of anti-IgE therapy, and tries to give a rational approach for future indications and directions of treatment with anti-IgE antibodies. PMID- 16248825 TI - Mucosal immunoregulation: transcription factors as possible therapeutic targets. AB - Much progress has been recently made with regard to our understanding of the mucosal immune system in health and disease. In particular, it has been shown that uncontrolled mucosal immune responses driven by lymphocytes or non-lymphoid cells may lead to immunological diseases such as allergy, hypersensitivity and inflammation. Thus, a more detailed understanding of mucosal immune regulation and decision making at mucosal surfaces is essential for a better understanding of mucosal immune responses in health and disease. Antigen presenting cells and T lymphocytes play a key role in controlling mucosal immune responses. To deal with this key task, T helper cells differentiate into functionally distinct subsets: TH1 (CD4+ T Helper cells), TH2, TH3, Tr1, and CD4+CD25+ T (Treg) cells. This review summarizes the role of antigen presenting cells, eosinophils, mast cells and T-cell subsets in the pathogenesis of allergic inflammation and intestinal inflammation. Furthermore, we discuss novel immunological treatment modalities for allergic inflammation (e.g. allergic asthma) and chronic intestinal inflammation (e.g. inflammatory bowel diseases (IBD)) such as the control of the expression of transcription factors to redirect pathological immune responses. PMID- 16248826 TI - Prophylaxis and therapy of allergy by mucosal tolerance induction with recombinant allergens or allergen constructs. AB - The mucosal immune system, present along the respiratory, gastrointestinal and genitourinary tract, has to discriminate between harmful pathogens and innocuous antigens, such as food, airborne antigens or the commensal bacterial flora. Therefore the mucosal immune system has acquired two opposing immunological functions, i.e. the induction of immunity and defence of mucosal pathogens, and the induction and maintenance of tolerance to environmental antigens and bacterial flora. As described for autoimmunity a breakdown or failure of tolerance induction is believed to lead also to allergies and food enteropathies. Based on the physiological role to prevent hypersensitivity reactions, tolerance induction via the mucosa has been proposed as a treatment strategy against inflammatory diseases, such as allergies. The aim of our research is to develop mucosal allergy vaccines based on the induction of mucosal tolerance and/or the induction of counter-regulatory immune responses with or without the use of certain mucosal antigen delivery systems, such as lactic acid bacteria. The use of recombinant allergens instead of allergen extracts with varying allergen content and composition may be essential for improvement of the treatment efficacy. In the present review we give examples of different animal models of type I allergy/asthma. Using these models we demonstrate that recombinant allergens or hypoallergenic variants thereof can be successfully used to induce mucosal tolerance in a prophylactic as well as a therapeutic treatment regime. That the concept of mucosal tolerance induction/mucosal vaccine delivery may in principal also function in humans is supported by recent clinical trials with locally (sublingual) applied immunotherapy. PMID- 16248827 TI - Design of protective and therapeutic DNA vaccines for the treatment of allergic diseases. AB - The DNA vaccine revolution has opened a vast scope of novel approaches for protective and therapeutic treatments of type I allergy. This review gives an overview on the current status of allergy DNA vaccines and presents advances in the design of vaccine constructs. An immense number of concurring studies have proven the stimulation of Th1 cells and the induction of a balanced Th1/Th2 cytokine milieu as the fundamental mechanisms underlying the anti-allergic effects of DNA vaccines. Basic vaccine formulations thus can be optimized by improving the cellular immunogenicity via co-administration of cytokines, co expression or co-application of immunostimulatory DNA sequences or adapting the codon usage. The latter is a frequent and major reason for impaired vaccine expression (e.g. translation of plant allergen genes in mammal cells). Because of unwanted side effects during conventional specific immunotherapy with allergen extracts, safety is increasingly demanded for both, protein and DNA vaccines for allergy treatment. We discuss the creation of hypoallergenic DNA vaccines based on deliberate allergen gene fragmentation, the use of mutations and the routine production of hypoallergenic DNA vaccines by forced ubiquitination. Furthermore, allergen-expressing DNA replicon vaccines are introduced, which enable a drastic reduction of the vaccine dose without loss of anti-allergic efficacy. Finally, the development of DNA multi vaccines and fusion vaccines for protective and therapeutic applications against certain groups of allergens is addressed. PMID- 16248828 TI - Evidence of probiotics in prevention of allergy and asthma. AB - Previous research into the causes of allergic diseases was mostly focussed on potential risk factors in the environment, with little success, however. Over the past 10 years, focus has therefore more been directed against protective factors that could enhance the development of tolerance to allergens which were previously encountered early in life, but are now lost in modern affluent societies. In particular, the role of childhood infections has been discussed, but so far these studies have not been conclusive. Recent epidemiological studies and experimental research suggest that the microbial environment and exposure to microbial products in infancy modifies immune responses and enhances the development of tolerance to ubiquitous allergens. The intestinal microflora may play a particular role in this respect, as it is the major external driving force in the maturation of the immune system after birth and animal experiments have shown it to be a prerequisite for normal development of oral tolerance. The composition of the microflora differs between healthy and allergic infants and in countries with a high and low prevalence of allergies. These differences are apparent within the first week of life, or even in the maternal vaginal flora during pregnancy and thus precede clinical symptoms. The use of live microorganisms that might be beneficial to health has a long tradition and the safety is well documented. Prospective intervention studies, in which the gut flora was modified from birth have yielded encouraging results and may suggest a new mode of primary prevention of allergy in the future. PMID- 16248829 TI - Erythropoietin in heart failure and other cardiovascular diseases: hematopoietic and pleiotropic effects. AB - Erythropoietin is a hypoxia-induced hormone that is a major regulator of normal erythropoiesis. Over the last decade, the production of recombinant human erythropoietin has revolutionized the treatment of anemia associated with chronic renal failure, and has led to a greater understanding of anemia pathophysiology and to the elucidation of the interactions of erythropoietin, iron, and erythropoiesis. Anemia has been shown to be independently associated with increased mortality and disease progression. Potential survival benefits associated with correction of anemia have expanded considerably the indications of erythropoietin use in various patient populations and are leading to consideration of earlier, more aggressive treatment of mild to moderate anemia. The results of such treatment are promising in a variety of new clinical settings, including anemia associated with congestive heart failure. Furthermore, the erythropoietin receptor is widely distributed in the cardiovascular system, including endothelial cells, smooth muscle cells and cardiomyocytes and preclinical studies have established erythropoietin to be a pleiotropic cytokine with anti-apoptotic activity and tissue-protective actions in the cardiovascular system, beyond correction of hemoglobin levels. Despite some potential adverse effects, such as hypertension, and the occurrence of erythropoietin resistance, early studies in heart failure patients with anemia suggest that erythropoietin therapy is safe and effective in reducing left ventricular hypertrophy, enhancing exercise performance and increasing ejection fraction. Anemia is found in about one-third of all cases of congestive heart failure (CHF). The most likely common cause is chronic renal insufficiency, which is present in about half of all CHF cases. However, anemia can occur in CHF without renal insufficiency and is likely to be due to excessive cytokine production. The anemia itself can worsen cardiac function, both because it causes cardiac stress through tachycardia and increased stroke volume, and because it can cause a reduced renal blood flow and fluid retention, adding further stress to the heart. Long-standing anemia of any cause can cause left ventricular hypertrophy, which can lead to cardiac cell death through apoptosis and worsen CHF. Therefore, a vicious circle, cardio-renal anemia syndrome, is set up wherein CHF causes anemia, and the anemia causes more CHF and both damage the kidneys worsening the anemia and the CHF further and increasing mortality. There is now evidence that early correction of the CHF anemia with subcutaneous erythropoietin and intravenous iron improves shortness of breath and fatigue, cardiac function, renal function and exercise capacity, reducing the need for hospitalization and improving quality of life. In the present review we discuss the data on current clinical use of erythropoietin in cardiovascular disease, with the main focus on the treatment of congestive heart failure, and summarize the advances and progress made in the understanding of the hematopoietic and pleiotropic effects of erythropoietin in the cardiovascular system. PMID- 16248830 TI - Role of PPAR- gamma agonist thiazolidinediones in treatment of pre-diabetic and diabetic individuals: a cardiovascular perspective. AB - The peroxisome proliferator-activated receptors (PPARs) are nuclear fatty acid receptors, which contain a type II zinc finger DNA binding motif and a hydrophobic ligand binding pocket. These receptors are thought to play an important role in metabolic diseases such as obesity, insulin resistance, and coronary artery disease. Three subtypes of PPAR receptors have been described: PPARalpha, PPARdelta/beta, and PPARgamma. PPARalpha is found in the liver, muscle, kidney, and heart. In the liver, its role is to up-regulate genes involved in fatty acid uptake (beta-oxidation and omega-oxidation). PPARdelta/beta is involved in fatty acid oxidation in muscle. PPARgamma has high expression in fat, low expression in the liver, and very low expression in the muscle. The thiazolidinediones (TZD) are synthetic ligands of PPARgamma. By activating a number of genes in tissues, PPARgamma increases glucose and lipid uptake, increases glucose oxidation, decreases free fatty acid concentration, and decreases insulin resistance. There is a sound rationale for the use of TZDs in patients with type 2 diabetes mellitus and promising preliminary data in patients with patients with pre-diabetes. In patients with type 2 diabetes, thiazolidinediones had been shown to decrease mean HbA(1c)by 1.5% and lower HbA(1c) to less than 7% in 30% of patients. Decreased muscle insulin resistance primarily mediates the glucose lowering effect. In addition, there are several nonhypoglycemic effects of TZDs which may be beneficial to both diabetics and patients with pre-diabetes. These include effects on lipid metabolism, blood pressure, endothelial function, atherosclerotic plaque, coagulation, and albuminuria. In a pilot study, we recently demonstrated that insulin sensitizers such as thiazolidinediones appear to be associated with better clinical outcomes compared to insulin providers in diabetic patients presenting with acute coronary syndromes. In another study, we showed that the prediabetic state is a marker for worse prognosis in patients with acute coronary syndromes. In this article, we review the existing literature on the effectiveness of PPAR-gamma agonists in patients with either overt diabetes or a prediabetic state. PMID- 16248831 TI - The role of renin angiotensin system blockade in the treatment of atrial fibrillation. AB - Atrial fibrillation (AF) is the most common cardiac arrhythmia seen in clinical practice. The understanding of the pathophysiology of AF has changed drastically during the last several decades. Recent observations have challenged the concept of the multiple circuit reentry model in favor of single focus or single circuit reentry models. Atrial electrical dysfunction provides a favorable substrate and transmembrane ionic currents are key determinants. Recent research is focusing increasingly on the atrial structural remodeling, which underlies the development of AF in different pathological conditions. This has led to concepts about how interfering with the substrate might prevent AF development and recurrence. Particular interest has been generated in the role of renin angiotensin system (RAS) blockade in reversing the electrical and structural remodeling of diseased atria. The mechanisms for the preventive effect of angiotensin converting enzyme inhibitors (ACEi) or angiotensin-II (AT-II) type 1 receptor blockers (ARB) in AF are probably complex. They may comprise general haemodynamic changes leading to lower intra-atrial pressure and wall-stress, or reduce in atrial fibrosis, connexin43 over-expression and conduction delay. The promising results of several clinical trials concerning RAS blockade may herald a whole new era of AF treatment, where AF is prevented and treated by modifying its substrate rather than fighting it electrically. This review centers on the pathophysiology of the structural and electrical remodeling in AF, the possible mechanisms by which RAS blockade may reverse electrical and structural remodeling of diseased atria and on the role of ACEi or ARB blockers in AF prevention and treatment that has already been postulated both experimentally and clinically. PMID- 16248832 TI - Will this be the end of the anticoagulation clinic for patients with atrial fibrillation? AB - Atrial fibrillation is the most common clinical arrhythmia and with an ageing population, it is an increasing cause of hospital admissions, morbidity and mortality. The most feared complication of atrial fibrillation is stroke. A number of studies have demonstrated that warfarin is at least moderately effective at reducing thromboembolic risk in stroke yet its use in both the community and in secondary care is suboptimal. Concerns about drug interactions, frequent blood monitoring and the risks of over and under coagulation have led to under prescription. Direct thrombin inhibitors are under investigation as an alternative to warfarin for thromboembolic prophylaxis in atrial fibrillation. Two large studies (SPORTIF III and SPORTIF V) have recently been published examining the effectiveness of the direct thrombin inhibitor ximelagatran at reducing thromboembolic risk. Ximelagatran was shown to be non-inferior to warfarin for the prevention of thromboembolic complications. Concerns however have arisen about long-term safety, particularly the possible effects on hepatic function. This review examines the data and discusses whether the introduction of these drugs could result in the end of the anticoagulation clinic for patients with atrial fibrillation. PMID- 16248833 TI - Drug therapy in Brugada syndrome. AB - Sudden cardiac death in healthy individuals with structurally normal hearts and a characteristic morphology of the QRS complex resembling a right bundle branch block with elevation of the ST segment in V1 to V3 is known as Brugada syndrome (BrS). Although placement of an implantable cardioverter-defibrillator is considered the only effective therapy for symptomatic patients, some authors have repeatedly reported a beneficial effect of quinidine and isoproterenol in patients with BrS. Also, isolated case reports on the usefulness of cilostazol, sotalol, and mexiletine have been described. The present article reviews the mechanisms by which these drugs may act and their role in the pharmacotherapy of BrS. Other possible agents, mainly I(2) blockers, are also reviewed. PMID- 16248834 TI - An old drug with a new purpose: cardiovascular actions of acetaminophen (paracetamol). AB - For over 50 years, acetaminophen (paracetamol) has been a staple in industrialized and non-industrialized countries for the treatment of pain and fever. Although its precise mechanisms of action are not known, the drug generates dose-dependent reduction in circulating prostaglandins, inhibits myeloperoxidase and the oxidation of lipoproteins, and appears to confer cardioprotection by blocking the effects of hydroxyl radical, peroxynitrite, and hydrogen peroxide. The drug might inhibit cyclooxygenase, although its ultimate target(s) is (are) still unclear. Sadly, since most investigations of acetaminophen have focused on its analgesic/antipyretic properties and hepatotoxicity, the effects of the drug on other mammalian organ systems, including the heart and circulation, have been ignored. Recently, work in our laboratory has shown acetaminophen to have a protective role in the injured mammalian myocardium. The cardioprotection was first observed in isolated, perfused guinea pig hearts subjected to ischemia-reperfusion injury. Hearts pretreated with acetaminophen recovered greater ventricular function and exhibited improved myofibrillar ultrastructure when compared to vehicle-treated hearts. More recent in vitro investigations have suggested protective roles for acetaminophen in barbiturate-induced arrhythmogenesis and myocardial hypoxia reoxygenation injury. We have also extended our work to the in vivo arena. There, we found that acetaminophen reduced infarct size in dogs exposed to 60 minutes regional myocardial ischemia and 180 minutes reperfusion. We invite and encourage readers of this review to repeat/duplicate our experiments. Such work is needed to either challenge or support our findings. Further, more clinically-relevant work depends on these basic and related translational experiments. PMID- 16248835 TI - Effect of omeprazole on the hydroxylation of warfarin enantiomers in human: in vitro studies with liver microsomes and cDNA-expressed cytochrome P450 isozymes. AB - Clinically observed warfarin-omeprazole interaction has been found to be associated with the inhibition of R-warfarin hydroxylation by omeprazole. The present study was conducted in human liver microsomes and cDNA-expressed cytochrome P450s to assess the inhibitory potential of omeprazole on the hydroxylation of warfarin enantiomers, and to identify the cytochrome P450 isozymes involved in the inhibition of hydroxylation of warfarin enantiomers by omeprazole, and to evaluate the extent to which the in vitro data is predictive of the actual pharmacokinetic interaction between warfarin and omeprazole observed in vivo. Omeprazole inhibited the formation of R-6-, R-7- and S-7 hydroxywarfarin with the Ki values of 40, 22 and 116 microM, respectively. Its inhibitory effect was selective towards R-warfarin. Further study conducted in cDNA-expressed cytochrome P450s (CYPs) demonstrates that the inhibition of the in vitro biotransformation of warfarin enantiomers by omeprazole is attributed to its inhibitory effect on the activities of CYP1A2, CYP3A4, CYP2C9 and CYP2C19. The extent of the in vivo warfarin-omeprazole interaction was underestimated as based on the Ki values obtained from the in-vitro inhibition study, suggesting an underestimation of the effective concentration of the inhibitor at the site of interaction or some other mechanisms involved in the drug interaction between warfarin and omeprazole. PMID- 16248836 TI - Cytochrome p450 enzymes mechanism based inhibitors: common sub-structures and reactivity. AB - The inhibition of human cytochrome P450s (CYPs) is one of the most common mechanisms which can lead to drug-drug interactions. The inhibition of CYPs can be reversible (competitive or non-competitive) or irreversible. Irreversible inhibition usually derives from activation of a drug by CYPs into a reactive metabolite, which tightly binds to the enzyme active site, leading to a long lasting inactivation. This process is called "mechanism based inhibition" or "suicide inhibition". The irreversible inactivation usually implies the formation of a covalent bond between the metabolite and the enzyme, which can lead to hapten formation and can in some cases trigger an autoimmune-response. For these reasons it is of utmost importance to study the mechanism of the CYP inhibition of new potential drugs as early as possible during the drug discovery process. The literature on CYPs is vast and covers numerous aspects of their biology and biochemistry, however to our knowledge there is no general and systematic review focusing on mechanism-based inhibitors; we have reviewed the literature and compiled all the available data on chemical entities, which are known to be CYP suicide inhibitors. Each compound is reported together with its chemical structure, the CYP isoform and the parameters describing the inactivation. Literature references are reported together with their PMID (PubMed ID number) to allow a fast retrieval of the papers. This review offers a quick reference to help predict liabilities of new chemical entities without carrying out extensive in vitro work, and will hopefully help in designing safer drugs. PMID- 16248837 TI - Coupling of conjugating enzymes and efflux transporters: impact on bioavailability and drug interactions. AB - Abstract: Conjugating enzymes are traditionally recognized as one of the major biological barriers to the entry of xenobiotics/drugs into systemic circulation and represent one of the main pathways for their elimination. Similar to drugs that undergo extensive phase I metabolism, drugs that undergo extensive conjugation have poor bioavailability and are more prone to metabolism-based drug interactions. Previously, enterohepatic recycling is used to explain why certain xenobiotics have half-lives that are much longer than expected from intravenous injection studies. In addition, changes in expression levels of metabolic enzymes due to chemical induction or suppression are often recognized as the source of drug interaction or toxicity of pollutants and carcinogens. These traditional approaches, whereas yielding highly valuable information, fail to recognize the fact that many conjugates (especially hydrophilic ones) cannot permeate the cell membrane. In the present review, we will focus on the coupling process that involves both conjugating enzymes and efflux transporters. We will briefly review conjugating enzymes capable of producing highly hydrophilic metabolic products. The other focus of this review is on various transporters capable of moving negatively charged hydrophilic conjugates across the cellular membrane. Evidence will support the hypothesis that efficient coupling of the conjugating enzymes and efflux transporters enables enterohepatic recycling and enteric recycling processes. Termed as a "revolving door" theory, the hypothesis focuses on the role played by efflux transporter capable of modulating the cellular excretion of hydrophilic metabolites. Coupling process in intestine, liver and kidney will be discussed with an emphasis on the intestinal coupling process, since we have just begun to understand it. Biological consequence and new insights into how coupling process can impact bioavailability of xenobiotics, biological functions of drugs and carcinogens, and drug interactions will be discussed. PMID- 16248838 TI - CYP1A1 is a major enzyme responsible for the metabolism of granisetron in human liver microsomes. AB - Granisetron, a potent 5-HT3 receptor antagonist, has been reported to be mainly metabolized to 7-hydroxygranisetron and a lesser extent to 9' desmethylgranisetron in humans. A previous study indicated that cytochrome P450 (CYP)3A4 is a major catalyst of 9'-demethylation, although the major CYP isoform(s) responsible for 7-hydroxylation are unknown. To clarify granisetron 7 hydroxylase, the in vitro metabolism of granisetron using expressed human CYPs and human liver microsomes was investigated. 7-Hydroxygranisetron was produced almost exclusively by CYP1A1, while, apparently, 9'-desmethylgranisetron was preferentially produced by CYP3A4. Marked inter-individual differences in the ratio of the formation of 7-hydroxygranisetron and 9'-desmethylgranisetron in human liver microsomes was observed. Granisetron 7-hydroxylase activity was strongly correlated with benzo[a]pyrene 3-hydroxylase activity (p<0.0001), but not with testosterone 6beta-hydroxylase activity in human liver microsomes. Furthermore, an anti-human CYP1A1 antibody completely inhibited 7-hydroxylation in human liver microsomes, however, the reaction was not inhibited at all by an anti-CYP3A4 antibody. On the other hand, granisetron 9'-demethylase activity correlated significantly not only with testosterone 6beta-hydroxylase activity (p<0.0001) but also with benzo[a]pyrene 3-hydroxylase activity (p<0.01). Consistent with this, both the anti-CYP1A1 and anti-human CYP3A4 antibodies inhibited the 9'-demethylase activity. These data indicate that CYP1A1 is a major enzyme responsible for the metabolism of granisetron via a main 7-hydroxylation pathway and an alternative 9'-demethylation route. This is the first report demonstrating the substantial contribution of CYP1A1 to the metabolism of a drug, although its role in the metabolism of environmental compounds is well established. PMID- 16248839 TI - Effect of some biologically interesting substituted tetrahydro-1,4-oxazines on drug metabolising enzymes and on inflammation. AB - The effect on hepatic drug metabolising enzymes was evaluated for three representative structures and that were selected from a series of substituted oxazine derivatives designed to possess particular pharmacological properties such as analgesic, antioxidant and hypolipidemic activity. In addition, since xenobiotic metabolism, reactive oxygen and nitrogen species, atherosclerosis and inflammation are interrelated and mutually affected, the effects of and on acute inflammation in vivo and lipoxygenase activity in vitro were also investigated. It was found that treatment of rats with caused induction of cytochrome P450, enhancement of the metabolism of aminopyrine in vitro and of zoxazolamine and hexobarbital in vivo. Compound appeared to induce particularly erythromycin N demethylation, while, a nitric ester, reduced the catalytically active cytochrome P450, although it increased the metabolism of specific cytochrome P450 substrates, i.e. 4-nitrophenol and erythromycin. Compounds and with strong hypolipidemic and antioxidant properties, reduced acute inflammatory response in two inflammation models and inhibited lipoxygenase activity in vitro. These results are helpful in optimising the biological profile as well as the potential applications of substituted oxazines. PMID- 16248840 TI - Metabolism and disposition of the antiviral nucleoside analogue AM365 in the isolated perfused rat liver. AB - The present study was designed to investigate the hepatic disposition of the prodrug AM365 and the generated antiviral guanosine analogue, AM188 in the isolated perfused rat liver (IPL). The livers of rats (n=12) were isolated and perfused with Krebs-Henseleit pH 7.4 buffer to which AM365 was added as a bolus to achieve an initial perfusate concentration of 22.4 micromol/L. During the 120 min period after administration of AM365, bile was collected in 10-min intervals and perfusate was collected at the mid-point of these intervals. Concentrations of AM365 and AM188 in perfusate and bile were quantified by HPLC. Following administration of AM365, its concentration in perfusate declined and the concentration of AM188 increased; the sum of the molar concentrations remained constant. The clearance and hepatic extraction ratio of AM365 were 3.3+/-2.4 mL/min and 0.110+/-0.079, respectively. The cumulative amount of AM365 excreted in bile during the 120-min perfusion period was approximately 0.21% of the bolus dose, and 0.36% of the total amount of AM365 cleared by the liver during the period. The cumulative amount of AM188 excreted in bile was about 0.48% of the total amount of AM188 formed during the perfusion period. In conclusion, AM365 was metabolised to AM188, which appeared to be the only metabolite and was not further biotransformed. The biliary excretion of AM365 and AM188 was negligible. PMID- 16248841 TI - Mechanisms of male infertility: role of antioxidants. AB - Defective sperm function is the most common cause of infertility, and until recently, was difficult to evaluate and treat. Mammalian spermatozoa membranes are rich in poly unsaturated fatty acids and are sensitive to oxygen induced damage mediated by lipid peroxidation. Hence, free radicals and reactive oxygen species [ROS] are associated with oxidative stress and are likely to play a number of significant and diverse roles in reproduction. The excessive generation of reactive oxygen species by abnormal spermatozoa and by contaminating leukocytes [leukocytospermia] has been identified as one of the few defined etiologies for male infertility. Moreover, environmental factors, such as pesticides, exogenous estrogens, and heavy metals may negatively impact spermatogenesis since male sperm counts were declined. In addition, aging is also likely to further induce oxidative stress. Limited endogenous mechanisms exist to reverse these damages. In a normal situation, the seminal plasma contains antioxidant mechanisms which are likely to quench these ROS and protect against any likely damage to spermatozoa. However, during genitourinary infection/inflammation these antioxidant mechanisms may downplay and create a situation called oxidative stress. Assessment of such oxidative stress status [OSS] may help in the medical treatment of male infertility by suitable antioxidants. The cellular damage in the semen is a result of an improper balance between ROS generation and scavenging activities. Therefore, numerous antioxidants such as vitamin C, vitamin E, glutathione, and coenzyme Q10, have proven beneficial effects in treating male infertility. A multi-faceted therapeutic approach to improve male fertility involves identifying harmful environmental and occupational risk factors, while correcting underlying nutritional imbalances to encourage optimal sperm production and function. PMID- 16248842 TI - Utility of recombinant cytochrome p450 enzymes: a drug metabolism perspective. AB - An important role of human cytochrome P450s (P450s) has been well recognized in the area of drug metabolism and pharmacokinetics. It has become possible in recent years to express catalytically active forms of these enzymes in various host systems. The resulting recombinant human P450s are either purified for studies of protein structure and the mechanism of catalysis or isolated in microsomal forms to serve the purposes of P450 phenotyping, metabolic stability screening and inhibitory potential evaluation. Intact mammalian cells expressing human enzymes may also be used to test the mutagenic and toxicity potential of drug candidates. The issue remains, however, that the data derived from recombinant P450s are not always consistent with those generated from human tissue preparations. The aim of this communication is to discuss applications of recombinant P450s in the drug discovery and development setting, with an emphasis on comparison of recombinant and human liver microsomal systems. PMID- 16248844 TI - The role of apoptotic pathways in Alzheimer's disease neurodegeneration and cell death. AB - Neuronal loss is associated with Alzheimer's disease (AD). However, it not clear what type of mechanisms underlie this neuronal loss and if neuronal loss is directly responsible for the progressive dementia of AD. This review summarizes the recent evidence for neuronal loss in AD relative to the level of cognitive impairment. It further describes the current evidence for an apoptotic mechanism in AD. Lastly, a summary of the evidence for synaptic loss being responsible for dementia rather than neuronal loss is presented. A novel hypothesis emerges from this data to explain all aspects of AD pathophysiology. This all inclusive hypothesis called the attrition hypothesis states that activation of the effector caspase-6 in AD due to one or a variety of insults is responsible for the breakdown of the cytoskeletal structure of neurites and damages proper trafficking of proteins and organelles thus resulting in the observed clinical and pathological features of AD. PMID- 16248845 TI - Oxidative stress: the old enemy in Alzheimer's disease pathophysiology. AB - The complex nature and genesis of oxidative damage in Alzheimer disease can be partly answered by mitochondrial and redox-active metal abnormalities. By releasing high levels of hydrogen peroxide, dysfunctional mitochondria propagate a series of interactions between redox-active metals and oxidative response elements. In the initial phase of disease development, amyloid-beta deposition and hyperphosphorylated tau may function as compensatory responses and downstream adaptations to ensure that neuronal cells do not succumb to oxidative injuries. However, during the progression of the disease, the antioxidant activity of both agents evolves into pro-oxidant activity representing a typical gain-of-function transformation, which can result from an increase in reactive species and a decrease in clearance mechanisms. PMID- 16248846 TI - Diversity of senile plaques in Alzheimer's disease as revealed by a new monoclonal antibody that recognizes an internal sequence of the Abeta peptide. AB - In order to have more specific tools available to approach amyloidogenesis in Alzheimer's disease (AD), we have produced several polyclonal and monoclonal antibodies that recognize specific sequences of the amyloid beta (Abeta) peptide. Here we present results that demonstrate that our monoclonal antibody EM5 recognizes an internal sequence (residues 11-16) of the Abeta peptide. This strategic localization of the epitope allowed us to employ this antibody, together with two previously reported polyclonal antibodies (EM2 and EM3, specific for AbetaX-40 and AbetaX-42, respectively), in an immunohistochemical study aimed at exploring the differential distribution of longer (AbetaX-40/42) and shorter (Abeta17-X) peptides along the various types of amyloid deposits of AD. This antibody panel was used in six AD brains, on sections from associative neocortex, striatum and cerebellar cortex. Single and double immunostaining revealed specific staining of vascular amyloid deposits and neuritic plaques by EM5 antibody, with high co-localization of EM2. Our results suggest that EM5 antibody recognizes pathogenic forms of Abeta deposits (amyloid angiopathy and neuritic plaques) and reveals the existence of a subset of plaques with a profile similar to vascular deposits. Additionally, our results show that diffuse plaques in AD brains may contain Abeta17-X peptides as its principal component. EM5 may be a useful tool in research both on human and transgenic mice tissue that may aid in the study of molecular heterogeneity of plaques in AD. PMID- 16248847 TI - Chitin-like polysaccharides in Alzheimer's disease brains. AB - The role of polysaccharides in the pathogenesis of Alzheimer disease (AD) is unclear. However, in light of studies indicating impaired glucose utilization in AD and increased activation of the hexosamine pathway that is seen with hyperglycemia, in the brains of patients with AD, aberrantly high levels of glucosamine may result in synthesis of glucosamine polymers such as chitin, a highly insoluble polymer of N-acetyl glucosamine, linearized by beta1-4 linkages. To examine this further, we studied brain tissue at autopsy from subjects with sporadic and familial AD using calcofluor histochemistry. Calcofluor excites on exposure to ultraviolet light and exhibits a high affinity for chitin in vivo by interacting with beta1-4 linkages. Amyloid plaques and blood vessels affected by amyloid angiopathy showed bright fluorescence. Moreover, treatment with chitinase, followed by beta-N-acetyl glucosaminidase showed a decrease in calcofluor fluorescence. Since chitin is a highly insoluble molecule and a substrate for glycan-protein interactions, chitin-like polysaccharides within the brain could facilitate nucleation of amyloid proteins in various amyloidoses including AD. PMID- 16248848 TI - Semantic memory disorders in Alzheimer's disease: clues from semantic priming effects. AB - Semantic memory deficits are a common landmark in Alzheimer's disease, but the nature of these impairments remains to be clarified. The tests used to explore this memory system are not specific and involve cognitive processes often disturbed in Alzheimer's disease. A complementary way to investigate semantic memory in neuropsychology is by using the semantic priming paradigm. Here, semantic priming refers specifically to the modification of a stimulus processing behind the presentation of a related stimulus; it is a short-lived phenomenon considered an implicit measure of semantic memory integrity. However, semantic priming studies have yielded contradictory results in Alzheimer's disease, with authors reporting normal priming, less-than-normal priming, or increased priming effects (hyperpriming). The aim of this paper is to review the literature investigating semantic priming in Alzheimer's disease, and to discuss the contradictory results in the context of current models of semantic processing. For a clear comprehension of the semantic priming patterns in this pathology, we will precise the methodology used and the characteristics of the Alzheimer's disease patients examined. Besides, the surprising hyperpriming phenomenon - often observed in Alzheimer's disease at the early stage of dementia--will also be explained in detail. These results from semantic priming represent invaluable clues to widen our knowledge and conceptions about deterioration of the semantic memory in Alzheimer's disease. PMID- 16248849 TI - Disconnection of language and memory in semantic dementia: a comparative and theoretical analysis. AB - In this paper, we present an illustrative case of Semantic Dementia (SD) and we review the literature on this relatively rare progressive neurodegenerative disorder. After reviewing the clinical, neuroimaging, neuropathological, and genetic features of SD, we propose a theoretical framework that addresses features of SD and relates them to features of other well known neuropsychiatric syndromes. Our 'on-line / off-line disconnection' model seeks to conceptualize SD as a syndrome of disconnection between two large distributed cortical networks, namely, between those networks that subserve language function and those that subserve memory function. PMID- 16248850 TI - Measuring morphological and cellular changes in Alzheimer's dementia: a review emphasizing stereology. AB - From a clinical as well as a neuropathological point of view Alzheimer's disease (AD) has been the focus of intense research for more than three decades. Most studies to identify morphometric correlates with the declining cognitive function in normal aging and AD have employed semi-quantitative methods to assess neuropathological markers such as neurofibrillary tangles, senile plaques, neuronal, or glial cell densities, and neuron sizes. To this end, many cell counting methods have employed two-dimensional designs in single sections, yielding estimates of cell numbers either as neuron densities (number of cell profiles per area) or estimates of the size distribution of neuron profiles in columns vertical to the cortical surface. This approach gives rise to difficulties in interpretation because of the three-dimensional size, shape, and orientation of the counted cells, and the effect of shrinkage artifacts. Modern stereological techniques offer a more rigorous approach for quantifying neuropathological changes associated with aging and degenerative disease. However the stereological studies also suffer from the limitations of high biological variability in AD-type neuropathology, and the relative scarcity of autopsied brains from well-studied non-demented comparison subjects. As a result, the clinicopathological associations between neuropathology and indices of cognitive performance in aging and AD are not yet firmly established. The requirement for the proper description of morphologic neuropathology of AD is clear: any macroscopic or microscopic abnormalities, are subtle and must consequently be demonstrated reproducibly in well-controlled studies. In this review we try to evaluate which, if any, of the contemporary claims for morphometric brain abnormalities in AD can be said to be well established, with special emphasis placed on human stereological post-mortal studies. PMID- 16248851 TI - Anticholinesterase and pharmacokinetic profile of phenserine in healthy elderly human subjects. AB - OBJECTIVE: To evaluate the safety, maximum tolerated dose (MTD), pharmacokinetics (PK), and pharmacodynamics (PD) of the acetyl-selective anticholinesterase, phenserine tartrate, in healthy elderly subjects. METHODS: 32 healthy elderly volunteers received single oral doses of phenserine tartrate (5-20 mg). Physical and vital signs were monitored over the ensuing 24 hours. Analyses were performed on plasma samples to determine PK, and PD were assessed using an erythrocyte acetylcholinesterase (AChE) inhibition assay. RESULTS: No serious adverse events (AEs) occurred; the most common were headache and vomiting. The MTD of phenserine tartrate was 10 mg. The Cmax and AUC(0-24) of phenserine increased with dose, but neither were dose-proportional. Subjects receiving 10 mg of phenserine tartrate had a Cmax of 1.95 ng/mL at 1.5 hours, and the mean peak inhibition (Imax) of AChE was 26% (range: 18-34%) at 1.75 hours (tImax) following dosing. The half life of AChE inhibition (tI1/2) was 11 hours. Evaluation of PK/PD relationships suggested a linear correlation between plasma phenserine concentration and AChE inhibition in the blood. CONCLUSIONS: Phenserine tartrate was safe and well tolerated when administered as a single oral dose of either 5 mg or 10 mg. An increase in the severity and frequency of AEs occurred at the 20 mg dose level. PMID- 16248852 TI - Multiple functions of l0036 in the regulation of the pathogenicity island of enterohaemorrhagic Escherichia coli O157:H7. AB - Diarrhoeagenic enterohaemorrhagic Escherichia coli and enteropathogenic E. coli attach to human intestinal epithelium and efface brush-border microvilli, forming an A/E (attaching and effacing) lesion. These human pathogens are phenotypically similar to the mouse pathogen Citrobacter rodentium. Genetically, they all have a homologous set of virulent genes involved in the A/E lesion, and these genes are organized on a LEE (locus of enterocyte effacement), a pathogenicity island. This island comprises 41 specific open reading frames, of which most are organized at five operons, LEE1, LEE2, LEE3, LEE4 and tir (LEE5). The expression of the LEE genes is regulated in a complicated manner, and current knowledge is that there are at least two positive regulators, Ler (LEE-encoded regulator) and GrlA (global regulator of LEE activator), and one negative regulator, called GrlR (global regulator of LEE repressor). In enterohaemorrhagic E. coli, GrlA is encoded by l0043, whereas GrlR is encoded by l0044. Here we report a fourth regulatory gene located in LEE3, namely l0036. Its expression is tightly controlled. When overexpressed, this factor, named Mpc (multiple point controller), interacts with Ler and suppresses the expression of the LEE proteins. When the translation is not initiated or terminated before maturation, the type III secretion of effectors is completely abolished. Therefore, together with the fact that several cis elements reside in the region that l0036 spans, l0036 appeared to have multiple functions in the regulation of LEE expression. PMID- 16248853 TI - Dual promoters control the cell-specific expression of the human cell death inducing DFF45-like effector B gene. AB - CIDE-B [cell death-inducing DFF45 (DNA fragmentation factor 45)-like effector B] is a member of the CIDE family of apoptosis-inducing factors. The highly restricted pattern of expression of CIDE-B in the liver and spleen suggests that a mechanism exists for the tissue- and cell-specific regulation of transcription of this gene. We have analysed the promoters of the human CIDE-B gene, particularly the mechanism of cell-specific transcription. Expression of CIDE-B is driven by two promoters which are responsible for the synthesis of two types of transcript, and Sp1 and Sp3 are key regulators of basal transcription from both the upstream and the internal promoter, as indicated by EMSAs (electrophoretic mobility-shift assays) and site-directed mutagenesis. Bisulphite sequencing analysis demonstrated that the upstream promoter was hypermethylated in cells that did not express the long transcript of CIDE-B, but was hypomethylated in cells that expressed this transcript. Furthermore, methylation of this region in vitro reduced the promoter activity to approximately 5% of the control. Thus methylation at CpG sites in the upstream promoter region appeared to be important for cell-specific synthesis of the long transcript. By contrast, HNF4alpha (hepatocyte nuclear factor-4alpha) bound to the internal promoter and enhanced its activity. Moreover, the short transcript of CIDE-B gene was expressed in cells which do not normally express this transcript upon introduction of exogenous HNF4alpha, demonstrating the involvement of HNF4alpha in the cell-specific synthesis of the short transcript. Thus our analysis revealed a novel mechanism for the cell-specific transcription of the human CIDE B gene, which involves epigenetic and genetic control at separate respective promoters. PMID- 16248857 TI - Selection of medical students: an Australian perspective. PMID- 16248854 TI - Pyroglutamyl peptidase type I from Trypanosoma brucei: a new virulence factor from African trypanosomes that de-blocks regulatory peptides in the plasma of infected hosts. AB - Peptidases of parasitic protozoans are emerging as novel virulence factors and therapeutic targets in parasitic infections. A trypanosome-derived aminopeptidase that exclusively hydrolysed substrates with Glp (pyroglutamic acid) in P1 was purified 9248-fold from the plasma of rats infected with Trypanosoma brucei brucei. The enzyme responsible was cloned from a T. brucei brucei genomic DNA library and identified as type I PGP (pyroglutamyl peptidase), belonging to the C15 family of cysteine peptidases. We showed that PGP is expressed in all life cycle stages of T. brucei brucei and is expressed in four other blood-stream-form African trypanosomes. Trypanosome PGP was optimally active and stable at bloodstream pH, and was insensitive to host plasma cysteine peptidase inhibitors. Native purified and recombinant hyper-expressed trypanosome PGP removed the N terminal Glp blocking groups from TRH (thyrotrophin-releasing hormone) and GnRH (gonadotropin-releasing hormone) with a k(cat)/K(m) value of 0.5 and 0.1 s(-1) x microM(-1) respectively. The half-life of TRH and GnRH was dramatically reduced in the plasma of trypanosome-infected rats, both in vitro and in vivo. Employing an activity-neutralizing anti-trypanosome PGP antibody, and pyroglutamyl diazomethyl ketone, a specific inhibitor of type I PGP, we demonstrated that trypanosome PGP is entirely responsible for the reduced plasma half-life of TRH, and partially responsible for the reduced plasma half-life of GnRH in a rodent model of African trypanosomiasis. The abnormal degradation of TRH and GnRH, and perhaps other neuropeptides N-terminally blocked with a pyroglutamyl moiety, by trypanosome PGP, may contribute to some of the endocrine lesions observed in African trypanosomiasis. PMID- 16248855 TI - Evolutionarily conserved structural motifs in bacterial GST (glutathione S transferase) are involved in protein folding and stability. AB - The bacterium Proteus mirabilis expresses a cytosolic class beta glutathione S transferase (PmGST B1-1) that is part of a family of multifunctional detoxication enzymes. Like other cytosolic GSTs, PmGST B1-1 possesses two local structural motifs, an N-capping box and a hydrophobic staple motif, both of which are located between amino acids 151 and 156. The N-capping box consists of a reciprocal hydrogen bonding interaction of Thr152 with Asp155, whereas the hydrophobic staple motif consists of a hydrophobic interaction between Phe151 and Ala156. By contrast with other GSTs, PmGST B1-1 displays distinct hydrogen bond interactions in the N-capping box. In mammalian GSTs these structural elements are critical for protein folding and stability. To investigate the role played by these two motifs in a distantly related organism on the evolutionary scale, site directed mutagenesis was used to generate several mutants of both motifs in PmGST B1-1. All mutants were efficiently overexpressed and purified, but they were quite unstable, although at different levels, indicating that protein folding was significantly destabilized. The analysis of the T152A and D155G variants indicated that the N-capping box motif plays an important role in the stability and correct folding of the enzyme. The analysis of F151A and A156G mutants revealed that the hydrophobic staple motif influences the structural maintenance of the protein and is implicated in the folding process of PmGST B1-1. Finally, the replacement of Thr152 and Asp155, as well as Phe151 and Ala156 residues influences the catalytic efficiency of the enzyme. PMID- 16248856 TI - Transport of polyamines in Drosophila S2 cells: kinetics, pharmacology and dependence on the plasma membrane proton gradient. AB - Polyamine transport activities have been described in diverse multicellular systems, but their bioenergetic mechanisms and molecular identity remain unclear. In the present paper, we describe a high-affinity spermine/spermidine transport activity expressed in Drosophila S2 cells. Ion-replacement experiments indicate that polyamine uptake across the cell membrane is Na+-, K+-, Cl-- and Ca2+ independent, but pH-sensitive. Additional experiments using ionophores suggest that polyamine uptake may be H+-coupled. Pharmacological experiments show that polyamine uptake in S2 cells is selectively blocked by MGBG {methylglyoxal bis(guanylhydrazone) or 1,1'-[(methylethanediylidine)-dinitrilo]diguanidine} and paraquat (N,N-dimethyl-4,4'-bipyridylium), two known inhibitors of polyamine uptake in mammalian cells. In addition, inhibitors known to block the Slc22 (solute carrier 22) family of organic anion/cation transporters inhibit spermine uptake in S2 cells. These data and the genetic tools available in Drosophila will facilitate the molecular identification and further characterization of this activity. PMID- 16248859 TI - Paracetamol overdose and hepatotoxicity at a regional Australian hospital: a 4 year experience. AB - BACKGROUND: Paracetamol is a component of a number of drugs taken in overdose (OD). The influence of alcohol use (acute or chronic) on the presentation and clinical course of paracetamol OD is contentious. This study explores the relationship between paracetamol OD, alcohol consumption and clinical outcomes at a regional Australian hospital. AIMS: To determine the frequency, circumstances and outcomes of paracetamol OD presentations to a regional Australian general hospital over a 4-year period. METHODS: Medical records of patients admitted to the Ballarat Health Services (BHS) as a result of paracetamol OD between January 2000 and December 2003 were reviewed. Patient demographics, amount of paracetamol ingested, other drug coingestions, alcohol history, previous medication OD, clinical course and outcomes were recorded. RESULTS: Annual admissions resulting from paracetamol OD almost doubled during the 4 years studied. The risk of a repeat paracetamol OD was highest within 4 weeks of the initial OD. Alcohol, benzodiazepines and antidepressants were commonly coingested. The strongest predictor of severe hepatotoxicity was delayed or no N-acetyl cysteine treatment in patients consuming greater than 10 g of paracetamol or with toxic serum paracetamol levels. A history of alcohol consumption did not appear to worsen outcomes. PMID- 16248858 TI - Ethics committee reviews and mutual acceptance: a pilot study. AB - AIMS: To develop a human research ethics committee (HREC) mutual acceptance (MA) model, based on the National Health and Medical Research Council's guidelines. The MA model aims to facilitate aspects of multicentre research and decrease the time taken to finalise the HREC review process. METHODS: Four HREC (The Alfred Hospital, Austin Health, Peter MacCallum Cancer Centre and Melbourne Health) agreed to participate in a 13-month pilot project to evaluate the MA model. Evaluation included times from submission to approval and stakeholder surveys. RESULTS: Seventeen consecutive studies were submitted to the MA pilot project. Stakeholders agreed that the MA model was efficient and effective and that submission and review processes had improved, with a demonstrable reduction in the levels of duplication. There was a 27% improvement in approval times for multicentre studies. CONCLUSIONS: Our (MA) model resulted in clear improvements in HREC processes and timelines. Stakeholder acceptance was high. This model provides a framework for a broader program of MA. PMID- 16248860 TI - Sputum bacteriology in patients hospitalized with acute exacerbations of chronic obstructive pulmonary disease and concomitant pneumonia in Hong Kong. AB - AIM: To study the sputum microbiology of patients admitted to a teaching hospital with acute exacerbations of chronic obstructive pulmonary disease (AECOPD) with concomitant pneumonia. METHODS: This was a retrospective study in a tertiary university hospital in Hong Kong. All episodes of AECOPD with pneumonia, patient demographics, length of stay, sputum culture and radiological results admitted in the first half of the year 2000 were retrieved from hospital records. RESULTS: There were 118 patients (91 male) with 150 episodes of AECOPD with concomitant pneumonia. The age of the patients was 74.8 +/- 10.0 years. Positive routine sputum culture was detected in 45.6% of the patients. Among these hospital admissions, 8.7, 4.0 and 12.0%, respectively, required intensive care unit admissions, mechanical ventilation and non-invasive positive pressure ventilation. Haemophilus influenzae was the commonest organism found in sputum (26.0%), followed by Pseudomonas aeruginosa (5.5%), Streptococcus pneumoniae (3.4%), and Moraxella catarrhalis (3.4%). Mycobacterium tuberculosis was found in 1.5% of the admissions. The presence of P. aeruginosa in sputum was associated with a longer hospital length of stay (9.1 +/- 3.1 vs. 7.1 +/- 4.2 days, P = 0.03). CONCLUSION: In contrast to most other studies reporting S. pneumoniae as the most likely bacterial pathogen, H. influenzae was the commonest bacterium isolated in sputum in our patients with AECOPD and concomitant pneumonia. PMID- 16248861 TI - Why do acute myocardial infarction patients not call an ambulance? An interview with patients presenting to hospital with acute myocardial infarction symptoms. AB - BACKGROUND: Studies from overseas indicate that patients with acute myocardial infarction (AMI) symptoms often fail to use the emergency services as recommended, thereby depriving themselves from life-saving treatment in case of cardiac arrest and delaying the time to myocardial reperfusion in the presence of a coronary occlusion. AIMS: To compare patients brought in by ambulance to those not brought in by ambulance and to question why some patients do not use the emergency services when presenting to hospital with AMI symptoms. METHODS: Prospective interview and follow up of consecutive patients presenting with AMI symptoms to the emergency department of a tertiary hospital in a metropolitan area within a 1-month period. RESULTS: Of the 215 patients presenting to the emergency department, 113 (53%) arrived by private transportation. Sixty (53%) of these felt their symptoms did not warrant calling the ambulance, 17 (15%) had first consulted their general practitioner. The private transport group accounted for 28% of documented AMI. CONCLUSIONS: A large proportion of patients with AMI symptoms refrain from calling the emergency services because they do not consider themselves critically ill. Education programmes appear to be warranted because more appropriate use of emergency services will save lives. PMID- 16248862 TI - Are humans infected by Hantaviruses in Australia? AB - Hantavirus antibody-positive rodents have been found across Australia although, to date, there are no reports of infections in humans. This could be due to misdiagnosis clinically and/or inadequate laboratory technique/skills. There are close trading ties between Australia and Asian countries as well as our geographical neighbours where both human and rodent infections are found, so importation is a continuing threat. We consider that further sero-epidemiological surveys are warranted among rodents (especially those captured from ports in Australia), in patients from renal and respiratory wards of hospitals, and in residents and employees close to harbours using more specific and sensitive laboratory techniques than have been available in the past. PMID- 16248863 TI - 'On being both professional and human': one woman's journey. PMID- 16248864 TI - Climatic, high tide and vector variables and the transmission of Ross River virus. AB - This report assesses the impact of the variability in environmental and vector factors on the transmission of Ross River virus (RRV) in Brisbane, Australia. Poisson time series regression analyses were conducted using monthly data on the counts of RRV cases, climate variables (Southern Oscillation Index and rainfall), high tides and mosquito density for the period of 1998-2001. The results indicate that increases in the high tide (relative risk (RR): 1.65; 95% confidence interval (CI): 1.20-2.26), rainfall (RR: 1.45; 95% CI: 1.21-1.73), mosquito density (RR: 1.17; 95% CI: 1.09-1.27), the density of Culex annulirostris (RR: 1.25; 95% CI: 1.13-1.37) and the density of Ochlerotatus vigilax (RR: 2.39; 95% CI: 2.30-2.48), each at a lag of 1 month, were statistically significantly associated with the rise of monthly RRV incidence. The results of the present study might facilitate the development of early warning systems for reducing the incidence of this wide-spread disease in Australia and other Pacific island nations. PMID- 16248865 TI - Survey of the Royal Australasian College of Physicians examination candidates: preparation and impact on lifestyle. AB - Candidates currently view the Royal Australasian College of Physicians' written examination as a major undertaking, knowing that approximately one-third fail on their first attempt. We anonymously surveyed New Zealand registrars who sat the written examination in 2004. We found that the majority of candidates spend long hours preparing over a several months, and that the examination had a detrimental impact on their personal lives. The results of this survey have important implications in the context of efforts to reduce the stress of doctors, and should lead to a change in either the examination itself or an increase in support provided to registrars preparing for it. PMID- 16248866 TI - Carotid sinus hypersensitivity as the initial manifestation of Takayasu arteritis. PMID- 16248867 TI - Combining treprostinil and sildenafil in the treatment of pulmonary hypertension. PMID- 16248868 TI - Cerebral nocardiosis. PMID- 16248869 TI - Multiple competing interests surround ethical review of clinical trials. PMID- 16248870 TI - Cystic macroprolactinoma: primary medical treatment? PMID- 16248871 TI - Nocardia infection following bone marrow transplantation. PMID- 16248875 TI - Approaches to mentoring: how to mentor and be mentored. PMID- 16248874 TI - Health care for the homeless assesses the use of adapted clinical practice guidelines. AB - This article describes a process of evaluating and adapting existing clinical practice guidelines (CPGs) for homeless individuals by different healthcare providers in multiple healthcare settings across the country. Data were collected using a standardized evaluation tool in nine sites across the United States. Clinicians completed an evaluation of the CPG after every use. Most clinicians used the CPG five times. Descriptive statistics were reported on the characteristics of the clinicians, and the utility of the guidelines and written comments. Clinicians had an average of 12 years of clinical experience, 8 years of which were specifically spent working with homeless individuals. Ninety-one percent of the clinicians practiced in urban settings. The majority of clinicians felt the adapted guidelines met evaluation criteria. The major weaknesses reported the delineation of outreach and case management activities. Results did not vary by clinicians' disciplines, years of experience, or any other indicators. Clients and clinicians providing primary care to homeless individuals may benefit from utilizing Health Care for the Homeless Clinicians' Network adapted CPGs to assure quality, evidenced-based care to a vulnerable population. PMID- 16248873 TI - The new higher level classification of eukaryotes with emphasis on the taxonomy of protists. AB - This revision of the classification of unicellular eukaryotes updates that of Levine et al. (1980) for the protozoa and expands it to include other protists. Whereas the previous revision was primarily to incorporate the results of ultrastructural studies, this revision incorporates results from both ultrastructural research since 1980 and molecular phylogenetic studies. We propose a scheme that is based on nameless ranked systematics. The vocabulary of the taxonomy is updated, particularly to clarify the naming of groups that have been repositioned. We recognize six clusters of eukaryotes that may represent the basic groupings similar to traditional "kingdoms." The multicellular lineages emerged from within monophyletic protist lineages: animals and fungi from Opisthokonta, plants from Archaeplastida, and brown algae from Stramenopiles. PMID- 16248876 TI - Primary care management of patients following bariatric surgery. AB - PURPOSE: To evaluate the nutritional, psychosocial, and other primary care issues faced by nurse practitioners (NPs) and their patients in the long-term management of the increasing population of patients who have had bariatric surgery. DATA SOURCES: An extensive review of the literature provides the foundation for development of assessment and management strategies highlighted in a case study. CONCLUSIONS: Management of the patient after bariatric surgery does not end with successful surgical healing. Numerous long-term implications, including significant psychosocial and nutritional issues, require the informed attention of the primary care provider for the rest of the life span. IMPLICATIONS FOR PRACTICE: Each year an increasing number of obese patients pursue a surgical solution to obesity, up to an estimated 100,000 in 2004. Numerous long-term health implications are specific to this population. NPs can improve the quality of primary care to these patients by being informed regarding the different procedures and their impact on physiologic phenomena, and the psychosocial issues inherent to extreme weight loss. PMID- 16248877 TI - Addressing technology pros and cons. PMID- 16248878 TI - Achieving excellence in practice: a model for continuing education for nurse practitioners. AB - PURPOSE: To provide a continuing education (CE) model that will meet varied needs of nurse practitioners by incorporating the variable of years of experience in practice setting. DATA SOURCES: An empirical study conducted over a period of approximately 10 months, from Fall 2002 to Spring 2003 in one of the southeastern states. CONCLUSIONS: A model has been developed to plan and deliver CE education programs based on the changing CE needs of NPs as they gain experience and expertise. IMPLICATIONS FOR PRACTICE: Results of this study may be used to guide providers to develop and offer CE programs based on the CE needs of NPs related to length of time in clinical practice, general issues of practice, and specialty practice area needs. PMID- 16248879 TI - Educational options for acute care nurse practitioner practice. AB - PURPOSE: During the Acute Care Interest Group Forum held on June 18, 2005, at the American Academy of Nurse Practitioner's Conference, it was identified that information on educational options for acute care nurse practitioner (ACNP) practice was needed, particularly for NPs who may have had other educational preparation but who were now working in acute care practice settings. As a result, a workgroup was formed to compile information on educational options for ACNP practice, including post-master's programs. DATA SOURCES: Web site descriptions of ACNP programs (n= 70) and, as needed, phone calls to ACNP programs to obtain information on program offerings including post-master's and distance-learning options. CONCLUSIONS: Educational options for ACNP practice include master's and post-master's programs, along with national certification. A growing number of educational programs offer post-master's options as well as distance-learning formats for coursework. IMPLICATIONS FOR PRACTICE: NPs interested in obtaining ACNP education, including those with other NP specialty education who are now working in acute care settings, have a number of choices for pursuing ACNP practice. NPs working in acute care need to ensure that their educational preparation meets the requirements set forth by the scope of standards for ACNP practice and national requirements and state regulations. PMID- 16248880 TI - Knowledge, perceptions, and attitudes of advanced practice nursing students regarding medical genetics. AB - PURPOSE: To describe the current medical genetic knowledge and perceptions of graduate advanced practice nursing (advanced practice nurse [APN]/nurse practitioner and nurse anesthetist) students using survey data for future integration of genetic topics, principles, and healthcare issues into curriculum. DATA SOURCES: Survey data of APNs' perceived knowledge of genetics and a review of the literature from past research studies of students and current articles from professional journals and organizations. Web sites were those of the National Coalition of Health Professions for Education in Genetics and National Institutes of Health, Human Genome Research Institute; professional organizations; and the authors' professional, clinical, and educational experiences. CONCLUSIONS: Most APN students perceived they had minimum knowledge and prior training regarding medical genetics. There is a need to integrate genetic concepts, principles, and medical conditions into advanced practice nursing curriculum and to provide clinical experiences in genetic conditions across the life span and throughout the health and illness spectrum. APN students have positive attitudes toward integrating genetics into graduate curricula. Potential methods for program integration include readings, small group discussion, standardized patients, and role-play as measures to increase information. IMPLICATIONS FOR PRACTICE: The National Coalition for Health Profession Education in Genetics, the American Nursing Association, and the American College of Nursing Education have recommended integration of genetics knowledge and skills into routine health care to provide effective interventions for individuals and families. However, previous research and data from this study have revealed that many nurses have minimal training in genetics. Advanced practice nurses must be knowledgeable on genetic principles, topics, and the ethical, legal, and social implications related to medical genetics to increase the ability to diagnose, prevent, and treat diseases and to provide effective care for individuals and families. PMID- 16248881 TI - A study of midlife women's reasons for changing healthcare providers. AB - PURPOSE: The purpose of this study was to examine the reasons midlife women report for changing healthcare providers and to determine if there were any differences in reasons given for the change based on gender or ethnicity. DATA SOURCES: This was an analysis of data collected from a healthy community-based sample of midlife women as part of a longitudinal 5-year study of changes in health outcomes during transition to menopause. Women were queried about their experience in changing healthcare providers. CONCLUSIONS: Over 42% indicated that they had changed health providers because of dissatisfaction with care. The component accounting for the majority of the variance was related to communication issues. There were no significant differences across ethnic groups in decision to change providers or in reasons for their dissatisfaction with care. However, their reports of experiencing racism in the healthcare system were troubling. IMPLICATIONS FOR PRACTICE: The ability of the provider to communicate with women may have implications in women's choices in health care later in life. PMID- 16248882 TI - Barriers to coronary heart disease risk modification in women without prior history of coronary heart disease. AB - PURPOSE: To explore the extent to which women perceive barriers to coronary heart disease (CHD) risk modification and to determine if access to a nurse practitioner (NP) decreases perceived barriers to CHD risk modification. DATA SOURCES: Surveys completed by 120 women between the ages of 35 and 60 years, with no known history of CHD. The barriers scale was used to examine women's perceived barriers to CHD risk modification. CONCLUSIONS: Women with access to an NP had less perceived barriers to CHD risk modification. IMPLICATIONS FOR PRACTICE: NPs are ideally suited to decrease the mortality and morbidity associated with CHD through education strategies and attention to individual barriers women face when attempting to incorporate CHD risk factor modification into their lifestyles. PMID- 16248883 TI - Noradrenaline overflow in mouse dentate gyrus following locus coeruleus and natural stimulation: real-time monitoring by in vivo voltammetry. AB - The pattern of catecholaminergic innervation of the dentate gyrus (DG) of the hippocampus, particularly the relatively dense and selective noradrenergic input, creates favourable conditions for real-time monitoring of noradrenaline (NA) release following stimulation of the locus coeruleus (LC) by in vivo voltammetry. Two electrochemically active species with different temporal characteristics were registered in the DG following electrical stimulation of the LC. Several approaches, including testing of anatomical and pharmacological specificity, coating of microelectrodes with Nafion and use of fast cyclic voltammetry, were used to verify the characteristics of electrochemical responses. The first sharp peak that appeared immediately during stimulation was definitely associated with NA overflow. The second late peak was possibly attributable to ascorbic acid. We examined the characteristics of alpha-2 adrenoceptor regulation of NA release in the DG, and showed for the first time that noradrenergic terminals resemble dopaminergic terminals in their mechanisms of increasing the refilling rate of the readily releasable pool following stimulation repeated at short intervals. Amperometric registration of NA in the DG was complicated by interference with electrical activity of hippocampus. This interference could be used, after appropriate filtration, for simultaneous recording from the same microelectrode of NA release and electrical activity of the hippocampus. PMID- 16248884 TI - 8-[2-(2-pentyl-cyclopropylmethyl)-cyclopropyl]-octanoic acid stimulates GABA release from interneurons projecting to CA1 pyramidal neurons in the rat hippocampus via pre-synaptic alpha7 acetylcholine receptors. AB - Nicotinic acetylcholine (ACh) receptors, such as alpha7, alpha3beta4 and alpha4beta2 receptors in the hippocampus, are suggested to modulate neurotransmitter release. 8-[2-(2-Pentyl-cyclopropylmethyl)-cyclopropyl]-octanoic acid (DCP-LA) (100 nM), a linoleic acid derivative, potentiated responses of alpha7, alpha3beta4 and alpha4beta2 ACh receptors expressed in Xenopus oocytes that are blocked by 3-(1-[dimethylaminopropyl] indol-3-yl)-4-[indol-3-yl] maleimide (GF109203X), a selective inhibitor of protein kinase C (PKC), except for alpha3beta4 ACh receptors. DCP-LA enhanced the nicotine-triggered release of GABA from rat hippocampal slices in the presence of tetrodotoxin in a bell-shaped dose-dependent manner at concentrations ranging from 10 nM to 10 microM, although DCP-LA by itself had no effect on GABA release. The DCP-LA action was inhibited by GF109203X or alpha-bungarotoxin, an inhibitor of alpha7 ACh receptors, but not by mecamylamine or dihydro-beta-erithroidine, an inhibitor of alpha3beta4 and alpha4beta2 ACh receptors. A similar effect on GABA release was obtained with 12 O-tetradecanoylphorbol 13-acetate, a PKC activator. DCP-LA (100 nM) also enhanced GABA release triggered by choline, an agonist of alpha7 ACh receptors, but not 3 [2(s)-azetidinylmethoxy] pyridine, an agonist of alpha4beta2 ACh receptors. In addition, DCP-LA (100 nM) increased the rate of nicotine-triggered GABA(A) receptor-mediated miniature inhibitory post-synaptic currents, monitored from CA1 pyramidal neurons of rat hippocampal slices, and the effect was also inhibited by GF109203X or alpha-bungarotoxin but not by mecamylamine. Thus, the results of the present study indicate that DCP-LA stimulates GABA release by enhancing activity of pre-synaptic alpha7 ACh receptors present on the GABAergic terminals of interneurons that transmit to CA1 pyramidal neurons via a PKC pathway. PMID- 16248885 TI - Prevention of mitochondrial dysfunction in post-traumatic mouse brain by superoxide dismutase. AB - Reactive oxygen species (ROS) are known to be involved in the pathogenesis of traumatic brain injury (TBI). Previous studies have shown that the susceptibility of mice to TBI-induced formation of cortical lesion is determined by the expression levels of copper-zinc and manganese superoxide dismutase (CuZnSOD and MnSOD, respectively). However, the underlying biochemical mechanisms are not understood. In this study, we measured the efficiency of mitochondrial respiration in mouse brains with altered expression of these two enzymes. While controlled cortical impact injury (CCII) with a deformation depth of 2 mm caused a drastic decrease in NAD-linked bioenergetic capacity in brain mitochondria of wild-type mice, the functional decrease was not observed in brains of littermate transgenic mice overexpressing CuZnSOD or MnSOD. In addition, a 1 mm CCII greatly compromised brain mitochondrial function in mice deficient in CuZnSOD or MnSOD, but not wild-type mice. Inclusion of the calcium-chelating agent, EGTA, in the assay solution could completely prevent dysfunction of oxidative phosphorylation in all mitochondrial samples, suggesting that the observed impairment of mitochondrial function was a result of calcium overloading. In conclusion, our results imply that mitochondrial dysfunction induced by superoxide anion radical contributes to lesion formation in mouse brain following physical trauma. PMID- 16248887 TI - Disruption of postsynaptic GABA receptor clusters leads to decreased GABAergic innervation of pyramidal neurons. AB - We have used RNA interference (RNAi) to knock down the expression of the gamma2 subunit of the GABA(A) receptors (GABA(A)Rs) in pyramidal neurons in culture and in the intact brain. Two hairpin small interference RNAs (shRNAs) for the gamma2 subunit, one targeting the coding region and the other one the 3'-untranslated region (UTR) of the gamma2 mRNA, when introduced into cultured rat hippocampal pyramidal neurons, efficiently inhibited the synthesis of the GABA(A) receptor gamma2 subunit and the clustering of other GABA(A)R subunits and gephyrin in these cells. More significantly, this effect was accompanied by a reduction of the GABAergic innervation that these neurons received. In contrast, the gamma2 shRNAs had no effect on the clustering of postsynaptic alpha-amino-3-hydroxy-5 methyl-4-isoxazolepropionic acid (AMPA) receptors, postsynaptic density protein 95 (PSD-95) or presynaptic glutamatergic innervation. A gamma2-enhanced green fluorescent protein (EGFP) subunit construct, whose mRNA did not contain the 3' UTR targeted by gamma2 RNAi, rescued both the postsynaptic clustering of GABA(A)Rs and the GABAergic innervation. Decreased GABA(A)R clustering and GABAergic innervation of pyramidal neurons in the post-natal rat cerebral cortex was also observed after in utero transfection of these neurons with the gamma2 shRNAs. The results indicate that the postsynaptic clustering of GABA(A)Rs in pyramidal neurons is involved in the stabilization of the presynaptic GABAergic contacts. PMID- 16248886 TI - Suppression of protein kinase Cepsilon mediates 17beta-estradiol-induced neuroprotection in an immortalized hippocampal cell line. AB - Although estrogens are neuroprotective in a variety of neuroprotection models, the precise underlying mechanisms are currently not well understood. Here, we examined the role of protein kinase C (PKC) in mediating estrogen-induced neuroprotection in the HT-22 immortalized hippocampal cell line. The neuroprotection model utilized calcein fluorescence to quantitate cell viability following glutamate insults. 17beta-Estradiol (betaE2) protected HT-22 cells when treatment was initiated before or after the glutamate insult. The inhibition of PKC by bis-indolylmaleimide mimicked and enhanced betaE2-induced neuroprotection. In contrast, the inhibition of specific PKC isozymes (alpha and beta) by Go6976, inhibition of 1-phosphatidylinositol 3 kinase by wortmannin, or inhibition of protein kinase A by H-89, did not alter cell viability, suggesting a specific involvement of PKC in an isozyme-dependent manner. We further examined whether estrogen interacts with PKC in a PKC isozyme-specific manner. Protein levels and activity of PKC isozymes (alpha, delta, epsilon, and zeta) were assessed by western blot analysis and radiolabeled phosphorylation assays respectively. Among the isozymes tested, betaE2 altered only PKCepsilon; it reduced the activity and membrane translocation of PKCepsilon in a manner that correlated with its protection against glutamate toxicity. Furthermore, betaE2 reversed the increased activity of membrane PKCepsilon induced by glutamate. These data suggest that the neuroprotective effects of estrogens are mediated in part by inhibition of PKCepsilon activity and membrane translocation. PMID- 16248888 TI - The membrane environment of endogenous cellular prion protein in primary rat cerebellar neurons. AB - We studied the membrane environment of cellular prion protein in primary cultured rat cerebellar neurons differentiated in vitro. In these cells, about 45% of total cellular prion protein (corresponding to a 35-fold enrichment) is associated with a low-density, sphingolipid- and cholesterol-enriched membrane fraction, that can be separated by flotation on sucrose gradient. Biotinylation experiments indicated that almost all prion protein recovered in this fraction was exposed at the cell surface. Prion protein was efficiently separated from this fraction by a monoclonal antibody immuno-separation procedure. Under conditions designed to preserve lipid-mediated membrane organization, several proteins were found in the prion protein-enriched membrane domains (i.e. the non receptor tyrosine kinases Lyn and Fyn and the neuronal glycosylphosphatidylinositol-anchored protein Thy-1). The prion protein-rich membrane domains contained, as well, about 50% of the sphingolipids, cholesterol and phosphatidylcholine present in the sphingolipid-enriched membrane fraction. All main sphingolipids, including sphingomyelin, neutral glycosphingolipids and gangliosides, were similarly enriched in the prion protein-rich membrane domains. Thus, prion protein plasma membrane environment in differentiated neurons resulted to be a complex entity, whose integrity requires a network of lipid mediated non-covalent interactions. PMID- 16248889 TI - Knock-down of POSH expression is neuroprotective through down-regulating activation of the MLK3-MKK4-JNK pathway following cerebral ischaemia in the rat hippocampal CA1 subfield. AB - We investigated the expression and subcellular localization of the multidomain protein POSH (plenty of SH3s) by immunohistochemistry and western blot analysis, as well as its role in the selective activation of mixed-lineage kinases (MLKs) 3, MAP kinase kinase (MKK) 4, c-Jun N-terminal kinases (JNKs) and the c-Jun signalling cascade in the rat hippocampal CA1 region following cerebral ischaemia. Our results indicated that the cytosol immunoreactivity of POSH was strong in the CA1-CA3 pyramidal cell but weak in the DG granule cell of the rat hippocampus both in sham control and after reperfusion. Co-immunoprecipitation experiments showed that the interactions of MLK3, MKK4 and phospho-JNKs with POSH were persistently enhanced during the early (30 min) and the later reperfusion period (from 1 to 3 days) compared with sham controls. Consistently, MLK3-MKK4 JNK activation was rapidly increased with peaks both at 30 min and 3 days of reperfusion. Intracerebroventricular infusion of POSH antisense oligodeoxynucleotides (AS-ODNs) not only significantly reduced the protein level of POSH, markedly decreased its interactions with MLK3, MKK4 and phospho-JNKs, but also attenuated the activation of the JNK signalling pathway. In addition, infusion of POSH AS-ODNs significantly increased the neuronal density in the CA1 region at 5 days of reperfusion. Our results suggest that POSH might serve as a scaffold mediating JNK signalling activation in the hippocampal CA1 region following cerebral ischaemia, and POSH AS-ODNs exerts its protective effects on ischaemic injury through a mechanism of inhibition of the MLK3-MKK4-JNK signalling pathway, involving c-Jun and caspase 3 activation. PMID- 16248890 TI - Genomic profiling of the neuronal target genes of the plasticity-related transcription factor -- Zif268. AB - The later phases of neuronal plasticity are invariably dependent on gene transcription. Induction of the transcription factor Zif268 (Egr-1) in neurones is closely associated with many forms of functional plasticity, yet the neuronal target genes modulated by Zif268 have not been characterized. After transfection of a neuronal cell line with Zif268 we identified genes that show altered expression using high density microarrays. Although some of the genes identified have previously been associated with forms of neuronal plasticity, the majority have not been linked with neuronal plasticity or Zif268 action. Altered expression of a representative sample of the novel target genes was confirmed in Zif268-transfected PC12 neurones, and in in vitro and in vivo models of Zif268 associated neuronal plasticity. In particular, altered expression of the protease inhibitor Cystatin C and the chemokine Cxcl10 was observed in striatal tissue after haloperidol administration. Surprisingly, the group of identified genes is enriched for components of the proteasome and the major histocompatibility complex. Our findings suggest that altered expression of these genes following Zif268 induction may be a key component of long lasting plasticity in the CNS. PMID- 16248892 TI - Telomerase down-regulation does not mediate PC12 pheochromocytoma cell differentiation induced by NGF, but requires MAP kinase signalling. AB - Telomerase is a ribonucleoprotein complex that maintains chromosomal telomere homeostasis and underlies continuous renewal of stem cells and immortalization of neoplastic cells. Telomerase is down-regulated during cell differentiation, but the mechanisms of down-regulation are largely unknown. Here, we examined roles of mitogen-activated protein (MAP) kinase and phosphatidylinositol-3 (PI3) kinase signalling pathways in telomerase down-regulation triggered by nerve growth factor (NGF), and the role of telomerase down-regulation in NGF-induced neural differentiation in PC12 cells. We report that NGF-induced telomerase down regulation requires MAP kinase signalling. While mutations of all putative Akt phosphorylation sites in telomerase reverse transcriptase (TERT) has no effect on telomerase activity, inhibition of MAP kinase signalling by PD98059 or U0126 abolishes NGF-induced telomerase down-regulation in a concentration-dependent manner. Reversal of NGF-induced telomerase down-regulation by TERT overexpression does not prevent NGF-induced neural differentiation. Down-regulation of telomerase by silencing TERT gene expression does not trigger cell differentiation in the absence of NGF, nor enhances NGF-induced differentiation. Thus, telomerase, withdraws by a mechanism at TERT gene transcription level involving MAP kinase signalling while cells cease proliferation and undergo differentiation. The withdrawal of telomerase is not required to mediate NGF induced PC12 cell differentiation and re-establishment of telomerase activity at significant levels does not inhibit differentiation. PMID- 16248891 TI - The galanin-R2 agonist AR-M1896 reduces glutamate toxicity in primary neural hippocampal cells. AB - Galanin is a neuropeptide involved in a variety of biological functions, including having a strong anticonvulsant activity. To assess a possible role of galanin in modulation of glutamatergic synapses and excitotoxicity, we studied effects of a galanin receptor 2(3) agonist (AR-M1896) on several molecular events induced by glutamate administration in primary neural hippocampal cells. Exposure of cells, after 5 days in vitro, to glutamate 0.5 mM for 10 min caused morphological alterations, including disaggregation of beta-tubulin and MAP-2 cytoskeletal protein assembly, loss of neurites and cell shrinkage. When present in culture medium together with glutamate, 1 and 10 nM of AR-M1896 reduced these alterations. Moreover, AR-M1896 counteracted glutamate-induced c-fos mRNA and c Fos protein up-regulation after 30-150 min, and 24 h, respectively. Massive nuclear alterations (Hoechst 33258 staining), observed 24 h after glutamate exposure, were also antagonized by AR-M1896 (0.1-100 nM) in a dose-dependent manner. These findings indicate that galanin, probably mainly through its type 2 receptor, interferes with events associated with glutamate toxicity. PMID- 16248893 TI - Effectiveness of harm reduction programmes for injecting drug users in Dhaka city. AB - This paper provides a brief overview of the harm reduction programme for injecting drug users (IDU) of CARE, Bangladesh in Dhaka city and uses data from surveillance and a focussed research study on a cohort of IDU, to evaluate the programme. The harm reduction programme in Dhaka is run by CARE, Bangladesh and includes needle/syringe exchange, awareness raising on HIV/STI, abscess management, condom distribution and advocacy with different groups of people. The needle/syringe exchange programme (NEP) has been in place since 1998, the 2nd Generation Surveillance in Bangladesh is being conducted since 1998, and an in depth cohort study, started in 2002, is being conducted in two areas of Dhaka city with approximately 500 IDU under CARE's NEP who are being followed bi annually to assess risk behaviour, incidence of HIV, hepatitis C and syphilis. As the surveillance and the cohort study are both closely associated with the NEP of CARE, Bangladesh, these data can be used to monitor the NEP. PMID- 16248894 TI - Molecular systems biology at the crossroads: to know less about more, or to know more about less? AB - Systems biology is a rapidly evolving discipline that endeavours to understand the detailed coordinated workings of entire organisms, with the ultimate goal to detect differences between health and disease, or to understand how cells or entire organisms react to the environment. The editorial provides a critical evaluation of what molecular systems analysis can and cannot accomplish with existing methodologies, and how systems biology needs to merge with reductionism to yield a more comprehensive and mechanistically insightful model of a cell or organism. PMID- 16248895 TI - Regulation of soluble vascular endothelial growth factor receptor (sFlt-1/sVEGFR 1) expression and release in endothelial cells by human follicular fluid and granulosa cells. AB - BACKGROUND: During the female reproductive cycle, follicular development and corpus luteum formation crucially depend on the fast generation of new blood vessels. The importance of granulosa cells and follicular fluid in controlling this angiogenesis is still not completely understood. Vascular endothelial growth factor (VEGF) produced by granulosa cells and secreted into the follicular fluid plays an essential role in this process. On the other hand, soluble VEGF receptor 1 (sFlt-1) produced by endothelial cells acts as a negative modulator for the bioavailability of VEGF. However, the regulation of sFlt-1 production remains to be determined. METHODS: We analyzed the influence of human follicular fluid obtained from FSH-stimulated women as well as of human granulosa cell conditioned medium on sFlt-1 production in and release from human umbilical vein endothelial cells (HUVEC) in vitro. Soluble Flt-1 gene expression was determined by RT-PCR analysis, amount of sFlt-1-protein was quantified by Sandwich-ELISA. RESULTS: Human follicular fluid as well as granulosa cell-conditioned medium significantly inhibit the production of sFlt-1 by endothelial cells on a posttranscriptional level. Treatment of cultured granulosa cells with either hCG or FSH had not impact on the production of sFlt-1 inhibiting factors. We further present data suggesting that this as yet unknown sFlt-1 regulating factor secreted by granulosa cells is not heat-sensitive, not steroidal, and it is of low molecular mass (< 1000 Da). CONCLUSION: We provide strong support that follicular fluid and granulosa cells control VEGF availability by down regulation of the soluble antagonist sFlt-1 leading to an increase of free, bioactive VEGF for maximal induction of vessel growth in the ovary. PMID- 16248896 TI - Pneumomediastinum as initial presentation of paralytic rabies: a case report. AB - BACKGROUND: Rabies is readily diagnosed when it presents as the classic furious form. Paralytic and atypical forms can pose significant problems in diagnosis. Catastrophic incidents included 7 organ transplant recipients who died of rabies recently in United States and Germany. Although rabies remains top in the lists of differential diagnosis of encephalitis in rabies endemic area, its complication may divert physicians from making a relevant management. We encountered an unusual case of paralytic rabies who presented with spontaneous pneumomediastinum. CASE PRESENTATION: A young male presented with fever and dysphagia. There was a history of fluctuating consciousness and aerophobia but they were absent or could not be demonstrated at the time of admission. He exhibited subcutaneous chest wall emphysema and was found to have pneumomediastinum which resulted in surgical intervention. He developed paralysis followed by seizures during postoperative period. Diagnosis was confirmed by demonstration of rabies RNA in saliva during the preterminal phase and by the autopsy. Over 200 hospital staff subsequently received rabies postexposure prophylaxis. CONCLUSION: Spontaneous pneumomediastinum can be a rare complication of rabies. It may lead clinicians to perform inappropriate treatment, particularly when phobic spasms are not present and agitation is not prominent. High level of awareness of rabies in any patient with confusion albeit subtle or with any obscure neurological presentations such as difficulty swallowing with no identifiable causes must be borne in mind. PMID- 16248897 TI - Adolescent distinctions between quality of life and self-rated health in quality of life research. AB - BACKGROUND: In adult quality of life (QOL) research, the QOL construct appears to differ from self-rated health status. Although increased QOL continues to be recognized as an important outcome in health promotion and medical intervention, little research has attempted to explore adolescent perceptual differences between self-rated health and QOL. METHODS: Correlational analyses were performed between self-rated health, physical health days and mental health days, and QOL. Data were collected from two different public high school adolescent samples during two different time periods (1997 & 2003) in two different geographic regions in the USA (a southern & midwestern state) with two different sample sizes (N = 5,220 and N = 140, respectively) using the CDC Youth Risk Behavior Survey (YRBS). The Centers for Disease Control and Preventions' health-related quality of life scale (HRQOL) provided estimates of self-rated health, physical health days and mental health days, and QOL. RESULTS: All correlation coefficients were significant in both samples (p < or = .0001), suggesting sample size was not a contributing factor to the significant correlations. In both samples, adolescent QOL ratings were more strongly correlated with the mean number of poor mental health days (r = .88, southern sample; r = .89, midwestern sample) than with the mean number of poor physical health days (r = .75, southern sample; r = .79, midwestern sample), consistent with adult QOL research. However, correlation coefficients in both samples between self-rated health and the mean number of poor physical health days was slightly smaller (r = .24, southern, r = .32, midwestern) than that between self-rated health and the mean number of poor mental health days (r = .25, southern, r = .39 midwestern), which is contrary to adult QOL research. CONCLUSION: Similar to adults, these results suggest adolescents are rating two distinct constructs, and that self-rated health and QOL should not be used interchangeably. QOL, in the context of public high school adolescents, is based largely upon self-reported mental health and to a lesser extent on self-reported physical health. Conversely, although self-reported mental health and self-reported physical health both contribute significantly to adolescent self-rated health, mental health appears to make a greater contribution, which is contrary to observations with adults. Health promoting efforts for adolescents may need to focus more on mental health than physical health, when considering population needs and type of micro or macro intervention. PMID- 16248898 TI - Out-of-home food outlets and area deprivation: case study in Glasgow, UK. AB - BACKGROUND: There is a popular belief that out-of-home eating outlets, which typically serve energy dense food, may be more commonly found in more deprived areas and that this may contribute to higher rates of obesity and related diseases in such areas. METHODS: We obtained a list of all 1301 out-of-home eating outlets in Glasgow, UK, in 2003 and mapped these at unit postcode level. We categorised them into quintiles of area deprivation using the 2004 Scottish Index of Multiple Deprivation and computed mean density of types of outlet (restaurants, fast food restaurants, cafes and takeaways), and all types combined, per 1000 population. We also estimated odds ratios for the presence of any outlets in small areas within the quintiles. RESULTS: The density of outlets, and the likelihood of having any outlets, was highest in the second most affluent quintile (Q2) and lowest in the second most deprived quintile (Q4). Mean outlets per 1,000 were 4.02 in Q2, 1.20 in Q4 and 2.03 in Q5. With Q2 as the reference, Odds Ratios for having any outlets were 0.52 (CI 0.32-0.84) in Q1, 0.50 (CI 0.31 0.80) in Q4 and 0.61 (CI 0.38 - 0.98) in Q5. Outlets were located in the City Centre, West End, and along arterial roads. CONCLUSION: In Glasgow those living in poorer areas are not more likely to be exposed to out-of-home eating outlets in their neighbourhoods. Health improvement policies need to be based on empirical evidence about the location of fast food outlets in specific national and local contexts, rather than on popular 'factoids'. PMID- 16248900 TI - Psychometric properties of the 25-item National Eye Institute Visual Function Questionnaire (NEI VFQ-25), Japanese version. AB - BACKGROUND: The importance of evaluating the outcomes of health care from the standpoint of the patient is now widely recognized. The purpose of this study is to develop and test a Japanese version of the National Eye Institute Visual Function Questionnaire (NEI VFQ-25). METHODS: A Japanese version was developed with a previously standardized method. The questionnaire and optional items were completed by 245 patients with cataracts, glaucoma, or age-related macular degeneration, by 110 others before and after cataract surgery, and by a reference group (n = 31). We computed rates of missing data, measured reproducibility and internal consistency reliability, and tested for convergent and discriminant validity, concurrent validity, known-groups validity, factor structure, and responsiveness to change. RESULTS: Based on information from the participants, some items were changed to 2-step items (asking if an activity was done, and if it was done, then asking how difficult it was). The near-vision and distance vision subscales each had 1 item that was endorsed by very few participants, so these items were replaced with items that were optional in the English version. For example, more than 60% of participants did not drive, so the driving question was excluded. Reliability and validity were adequate for all subscales except driving, ocular pain, color vision, and peripheral vision. With cataract surgery, most scores improved by at least 20 points. CONCLUSION: With minor modifications from the English version, the Japanese NEI VFQ-25 can give reliable, valid, responsive data on vision-related quality of life, for group-level comparisons or for tracking therapeutic outcomes. PMID- 16248901 TI - Implicit learning of non-spatial sequences in schizophrenia. AB - Recent studies have reported abnormal implicit learning of sequential patterns in patients with schizophrenia. Because these studies were based on visuospatial cues, the question remained whether patients were impaired simply due to the demands of spatial processing. This study examined implicit sequence learning in 24 patients with schizophrenia and 24 healthy controls using a non-spatial variation of the serial reaction time test (SRT) in which pattern stimuli alternated with random stimuli on every other trial. Both groups showed learning by responding faster and more accurately to pattern trials than to random trials. Patients, however, showed a smaller magnitude of sequence learning. Both groups were unable to demonstrate explicit knowledge of the nature of the pattern, confirming that learning occurred without awareness. Clinical variables were not correlated with the patients' learning deficits. Patients with schizophrenia have a decreased ability to develop sensitivity to regularly occurring sequences of events within their environment. This type of deficit may affect an array of cognitive and motor functions that rely on the perception of event regularity. PMID- 16248902 TI - Metamemory for faces following frontal lobe damage. AB - Previous research has provided evidence of metamemory impairments in patients with frontal lobe damage on verbal episodic memory tasks. In the present study, we employed metamemory paradigms to investigate whether patients with frontal lesions show monitoring deficits on semantic memory tasks involving facial stimuli. Patients with frontal lobe damage and healthy control subjects made memory decisions to famous faces in a retrospective confidence judgment task and in a prospective feeling-of-knowing (FOK) task. Results indicated that frontal patients performed worse than controls on the retrospective confidence task, but there were no differences between the groups on the FOK task. These findings suggest that metamemory deficits in frontal patients are not confined to specific stimulus domains (words vs. faces) or memory systems (episodic vs. semantic). In addition, the dissociation between retrospective confidence judgments and FOK accuracy documented in this study and also in a recent report by Schnyer et al. suggesting that metamemory should not be considered a unitary function with a single neuroanatomic substrate. PMID- 16248904 TI - Neuroanatomic correlates of CVLT-C performance following pediatric traumatic brain injury. AB - Traumatic brain injury (TBI) frequently results in memory problems, and the degree of memory impairment is related to injury severity and is commonly associated with lesions in frontal and temporal brain areas. This study examined the relationship among injury severity, brain lesions, and memory in children with moderate to severe TBI using Donders' (1999) 5-factor model of performance on the California Verbal Learning Test-Children's Version (CVLT-C). Seventy-six children underwent magnetic resonance imaging (MRI) scans 3 months post-TBI and testing 1 year post-TBI. Results showed injury severity (Glasgow Coma Scale) was not predictive of performance on 4 of the 5 factors. Volume of frontal and/or temporal brain lesions was significantly predictive of performance on 3 of the 5 factors. Unexpectedly, lesion volume outside these areas (extra-frontotemporal) was predictive of performance on all 5 factors. In contrast, Verbal IQ at 1 year was most strongly associated with preinjury factors (socioeconomic status and special education involvement), although extra-frontotemporal lesions also contributed to the variability in this measure. Results suggest that in children with moderate to severe TBI, extra-frontal/temporal lesions are predictive of memory outcome 1 year postinjury above and beyond initial severity or frontal/temporal contusions. This finding may relate to widespread diffuse axonal injury, which potentially disconnects brain circuits mediating memory following moderate to severe TBI. PMID- 16248903 TI - Affective facial and lexical expression in aprosodic versus aphasic stroke patients. AB - Past research has shown that lesions in the left cerebral hemisphere often result in aphasia, while lesions in the right hemisphere frequently impair the production of emotional prosody and facial expression. At least 3 processing deficits might account for these affective symptoms: (1) failure to understand the conditions that evoke emotional response; (2) inability to experience emotions; (3) disruption in the capacity to encode non-verbal signals. To better understand these disorders and their underlying mechanisms, we investigated spontaneous affective communication in right hemisphere damaged (RHD) stroke patients with aprosody and left hemisphere damaged (LHD) stroke patients with aphasia. Nine aprosodic RHD patients and 14 aphasic LHD patients participated in a videotaped interview within a larger treatment protocol. Two naive raters viewed segments of videotape and rated facial expressivity. Verbal affect production was tabulated using specialized software. Results indicated that RHD patients smiled and laughed significantly less than LHD patients. In contrast, RHD patients produced a greater percentage of emotion words relative to total words than did LHD patients. These findings suggest that impairments in emotional prosodic production and facial expressivity associated with RHD are not induced by affective-conceptual deficits or an inability to experience emotions. Rather, they likely represent channel-specific nonverbal encoding abnormalities. PMID- 16248905 TI - Assessment of spatial attention after brain damage with a dynamic reaction time test. AB - Lateralized spatial biases after brain damage are commonly assessed using batteries of paper-and-pencil tests. These tests hardly allow quantification of performance in different locations in space, and they tend to lose sensitivity along the course of recovery. We tested the dynamic Starry Night Test (SNT), a novel computerized test measuring reaction time and detection accuracy for visual target stimuli in a dynamic background, in 32 inpatients with right hemisphere stroke (RHS), 16 patients with left hemisphere stroke (LHS), and 9 healthy controls. As a group, only the RHS patients were significantly slower to respond to contralesional targets. Individually, 21 (66%) RHS patients and 5 (31%) LHS patients showed statistically significant contralateral deficits. In a number of RHS patients the SNT was more sensitive to the ipsilesional bias of spatial attention than the Behavioral Inattention Test (BIT), a standardized paper-and pencil test battery of unilateral spatial neglect. Two illustrative case reports show that the dynamic SNT, but not the BIT, was sensitive to the spatial deficit in recovered patients, one of whom was involved in repeated car accidents. The SNT overcomes serious shortcomings of paper-and-pencil tests of unilateral neglect. It provides a simple quantitative tool for monitoring the natural and treatment-induced recovery of patients. PMID- 16248899 TI - Epigenetics of cervical cancer. An overview and therapeutic perspectives. AB - Cervical cancer remains one of the greatest killers of women worldwide. It is difficult to foresee a dramatic increase in cure rate even with the most optimal combination of cytotoxic drugs, surgery, and radiation; therefore, testing of molecular targeted therapies against this malignancy is highly desirable. A number of epigenetic alterations occur during all stages of cervical carcinogenesis in both human papillomavirus and host cellular genomes, which include global DNA hypomethylation, hypermetylation of key tumor suppressor genes, and histone modifications. The reversible nature of epigenetic changes constitutes a target for transcriptional therapies, namely DNA methylation and histone deacetylase inhibitors. To date, studies in patients with cervical cancer have demonstrated the feasibility of reactivating the expression of hypermethylated and silenced tumor suppressor genes as well as the hyperacetylating and inhibitory effect upon histone deacetylase activity in tumor tissues after treatment with demethylating and histone deacetylase inhibitors. In addition, detection of epigenetic changes in cytological smears, serum DNA, and peripheral blood are of potential interest for development of novel biomolecular markers for early detection, prediction of response, and prognosis. PMID- 16248906 TI - Recall discriminability: utility of a new CVLT-II measure in the differential diagnosis of dementia. AB - Memory tests that are in a recall format have almost universally measured accuracy in terms of the number of target items reported by the examinee. However, this traditional scoring method can, in certain cases, result in artificially inflated memory accuracy scores. That is, just as a "yes" response bias and high false-positive rate on recognition testing can artificially inflate a patient's hit rate, so, too, a liberal response bias and high intrusion rate on recall testing can artificially inflate a patient's level of target recall. Recognition tests correct for this problem by using a discriminability measure that provides a single score of hit rate relative to false-positive rate; however, recall tests rarely provide a single score of recall accuracy that corrects for intrusion rate. In the present study, we examined the utility of a new recall discriminability measure that analyzes target recall relative to intrusion rate. Patients with Alzheimer's disease (AD) or Huntington's disease (HD) were administered the CVLT-II, which provides both the traditional measure of target recall and a new measure of recall discriminability. The results indicate that the new recall discriminability measure was superior to the traditional level of target recall measure in distinguishing the recall performance of AD and HD patients. Implications of these results for clinical practice and theories of memory disorder in dementia are discussed. PMID- 16248907 TI - A longitudinal study of confrontation naming in the "normal" elderly. AB - The longitudinal effects of age on confrontation naming using the 60-item Boston Naming Test (BNT) were studied in 541 "normal" elderly (ages 50-99). For participants with at least 4 annual assessments (n = 238), 150 were followed for > or =6 years, 81 for > or =8 years, and 43 for > or =10 years. A small practice effect (0.21 words, p = 0.06) and moderately high test-retest reliability were found when comparing the first 2 assessments, which were 9-15 months apart (r = 0.76, n = 353). Reliable change index scores indicated that an annual decline of > or =4 points on the BNT is needed for a statistically reliable decline in an individual. A gradient in the mean annual rate of change on the BNT was found with improvement in the 50s age group, no change in the 60s age group, and decline in the 70s and 80s age groups. When projected over 10 years, the magnitudes of the mean changes were relatively small, that is, a 1-word improvement for participants in their 50s and a 1.3-word decline for participants in their 70s. These findings demonstrate that lexical retrieval as measured by a visual object confrontation naming task is generally well preserved in aging with only subtle decline in the 7th and 8th decades of age. PMID- 16248908 TI - Effects of using same- versus alternate-form memory tests during short-interval repeated assessments in multiple sclerosis. AB - Repeated neuropsychological testing gives rise to practice effects in that patients become familiar with test material as well as test-taking procedures. Using alternate forms prevents the learning of specific test stimuli, potentially mitigating practice effects. However, changing forms could diminish test-retest reliability coefficients. Our objective was to examine test-retest effects in multiple sclerosis (MS) patients randomly assigned to same- (SF) or alternate form (AF) conditions. Thirty-four MS patients underwent neuropsychological evaluation. The battery included the California Verbal Learning Test II (CVLT-II) and the Brief Visuospatial Memory Test-Revised (BVMT-R), memory tests recommended by a recently convened consensus panel. Patients were randomly assigned to SF or AF groups and then tested at baseline and follow-up examination 1 week later. Analysis of variance tests (ANOVAs) revealed significant group x time interactions, with SF patients showing greater gain than AF patients. SF practice effects were often large, compromising test validity. Reliability coefficients were either equivalent or higher in the AF group, a finding attributed to ceiling effects and reduced variance in the SF group at retest. The generalizability of the findings may be limited to short test-retest intervals and the MS population. Nevertheless, I conclude that the use of CVLT-II and BVMT-R alternate forms likely helps preserve test validity without compromising test-retest reliability. PMID- 16248909 TI - Memory interference in multiple sclerosis. AB - To explore verbal memory impairments associated with multiple sclerosis (MS), we compared proactive and retroactive interference effects on the California Verbal Learning Test (CVLT; Delis et al., 1987) in a sample of 83 community-residing individuals with MS and 80 healthy participants. Individuals with MS demonstrated normal accumulation of proactive interference (PI), but attenuated release from PI relative to healthy individuals. Furthermore, accumulation of retroactive interference (RI) at short-delay free recall (SDFR) was intensified for those with MS as compared to healthy participants. Interestingly, accumulation of RI predicted long-term memory (LTM) only for participants with MS. These findings suggest that individuals with MS may experience particular difficulty when required to use semantic properties of information flexibly to facilitate verbal LTM. PMID- 16248910 TI - Workers' risk of unemployment after traumatic brain injury: a normed comparison. AB - We examined, among those persons working preinjury, the risk of unemployment 1 year after traumatic brain injury (TBI) relative to expected risk of unemployment for the sample under a validated risk-adjusted econometric model of employment in the U.S. population. Results indicate that 42% of TBI cases were unemployed versus 9% expected, relative risk (RR) = 4.5, 95% confidence interval (CI) (4.12, 4.95). The relative risk for unemployment was higher among males, those with higher education, persons with more severe injuries, and more impaired early neuropsychological or functional status. Difference in unemployment rates gave similar results for gender, severity of injury, and early neuropsychological and functional status. However, for education, the excess was smaller among those more highly educated, but the unemployment rate in the more highly educated in the general population was sufficiently small to yield a larger relative risk. In conclusion, after accounting for underlying risk of unemployment in the general population, unemployment is substantially higher after TBI for people who were employed when they were injured. The differential employment status varies depending on demographics, severity of brain injury, early functional outcome, and neurobehavioral indicators. For characteristics such as education, associated with rates of unemployment in the general population, different methods used to compare the rates may yield different results. PMID- 16248911 TI - Ecstasy (MDMA) exposure and neuropsychological functioning: a polydrug perspective. AB - Ecstasy (MDMA) is a popular drug that can act as a selective serotonin neurotoxin in several species. The goal of the present study was to examine the relationship between ecstasy exposure and cognitive functioning after controlling for other drug use and demographic variables. Furthermore, we assessed whether gender was a moderator of the relationship between cognitive functioning and ecstasy use. Data were collected from 31 men and 34 women with a wide range of ecstasy use (17 marijuana users with no ecstasy use and 48 ecstasy users ranging from low to heavy use). Participants were interviewed and administered a battery of neuropsychological tests. The primary finding was that ecstasy exposure was significantly related to poorer verbal learning and memory ability in a dose dependent manner, while no such relationship was observed between ecstasy exposure and executive functioning or attentional ability. Gender was found to significantly moderate the relationship between ecstasy consumption and design fluency. These results suggest primary memory dysfunction among abstinent recreational ecstasy users. This finding is consistent with reports of hippocampal vulnerability, particularly among heavy users. PMID- 16248912 TI - Neuropsychological deficits associated with driving performance in Parkinson's and Alzheimer's disease. AB - Neuropsychological and motor deficits in Parkinson's disease that may contribute to driving impairment were examined in a cohort study comparing patients with Parkinson's disease (PD) to patients with Alzheimer's disease (AD) and to healthy elderly controls. Nondemented individuals with Parkinson's disease [Hoehn & Yahr (H&Y) stage I-III], patients with Alzheimer's disease [Clinical Demetia Rating scale (CDR) range 0-1], and elderly controls, who were actively driving, completed a neuropsychological battery and a standardized road test administered by a professional driving instructor. On-road driving ability was rated on number of driving errors and a global rating of safe, marginal, or unsafe. Overall, Alzheimer's patients were more impaired drivers than Parkinson's patients. Parkinson's patients distinguished themselves from other drivers by a head turning deficiency. Drivers with neuropsychological impairment were more likely to be unsafe drivers in both disease groups compared to controls. Compared to controls, unsafe drivers with Alzheimer's disease were impaired across all neuropsychological measures except finger tapping. Driving performance in Parkinson's patients was related to disease severity (H&Y), neuropsychological measures [Rey Osterreith Complex Figure (ROCF), Trails B, Hopkins Verbal List Learning Test (HVLT)-delay], and specific motor symptoms (axial rigidity, postural instability), but not to the Unified Parkinson Disease Rating Scale (UPDRS) motor score. Multifactorial measures (ROCF, Trails B) were useful in distinguishing safe from unsafe drivers in both patient groups. PMID- 16248913 TI - Investigation of response inhibition in obsessive-compulsive disorder using the Hayling task. AB - This study investigates response inhibition deficits in obsessive-compulsive disorder (OCD) by using the Hayling task. Sixteen OCD washers, 16 OCD checkers, 16 social phobic patients and 16 nonanxious controls were asked to complete sentences with either the expected word (section A: "initiation") or an unrelated word (section B: "inhibition"). The groups did not differ in terms of section B minus section A latencies. However, OCD washers and checkers made significantly more errors (sentence-related responses) in section B than social phobic patients and controls. In the OCD patients, the frequency of these errors correlates with the total OCD severity score and the compulsion subscore, but not with the depression and anxiety scores. These findings suggest that OCD patients might present a specific deficit affecting the inhibition of a prepotent response. PMID- 16248914 TI - The use of word-reading to estimate "premorbid" ability in cognitive domains other than intelligence. AB - Diagnostic neuropsychological assessment requires the clinician to estimate a patient's premorbid abilities. Word reading tests, such as the National Adult Reading Test-Revised (NART-R), provide reasonably accurate estimates of premorbid IQ, but their capacity to benchmark other premorbid cognitive abilities remains unclear. In this extension of an earlier report, we administered the NART-R, an abbreviated Wechsler Adult Intelligence Scale (WAIS-R or WAIS-III), and 26 other cognitive measures to 322 reasonably healthy adults. While NART-R performance correlated robustly (rs > or = .72) with concurrent Verbal and Full Scale IQ, its correlation with all other cognitive measures was significantly lower. Thus, while it is appealing to use word reading as a proxy for premorbid functioning in other cognitive domains, the NART-R has limited utility for this because it does not predict current performance on other cognitive tests as well as it predicts IQ in healthy adults. PMID- 16248915 TI - Methodology is more important than statistics when determining reliable change. PMID- 16248916 TI - Anatomic substrates for, and function of, the functionally univentricular circulation before and after surgical procedures. PMID- 16248917 TI - Pathogenesis of congenital cardiac malformations and mechanisms of cardiac remodelling. PMID- 16248918 TI - The functionally univentricular circulation: anatomic substrates as related to function. PMID- 16248919 TI - Stem cells and cardiac hypertrophy. PMID- 16248920 TI - The neuroendocrinal system of the heart after construction of a Glenn anastomosis or the Fontan circulation. PMID- 16248921 TI - Cardiac and pulmonary physiology in the functionally univentricular circulation with reference to the total cavo-pulmonary connection. PMID- 16248922 TI - Ventricular dysfunction of the functionally univentricular heart: management and outcomes. PMID- 16248923 TI - Contractility-afterload mismatch after the Fontan operation. PMID- 16248924 TI - Cardiopulmonary response to exercise in patients with the Fontan circulation. PMID- 16248925 TI - Findings from new echocardiographic techniques concerning ventricular remodelling and function in the functionally univentricular heart. PMID- 16248926 TI - The role of magnetic resonance imaging in the evaluation of the functionally single ventricle before and after conversion to the Fontan circulation. PMID- 16248927 TI - Mathematical modelling of the impact of preoperative hypertrophy on the outcomes of completion of the Fontan circuit. PMID- 16248928 TI - Computational fluid dynamics in a model of the total cavopulmonary connection reconstructed using magnetic resonance images. PMID- 16248929 TI - Critical issues in studies of flow through the Fontan circuit after 10 years of investigation. PMID- 16248930 TI - Valveless pump models that laid a false but fortuitous trail on the way towards the total cavopulmonary connection. PMID- 16248931 TI - Effects of aerobic exercise training on the stiffness of central and peripheral arteries in middle-aged sedentary men. AB - Aortic pulse wave velocity (PWV) significantly decreased after 16 weeks of moderate-intensity exercise training (walking/jogging) in 17 sedentary middle aged men, whereas leg PWV did not. These results suggest that in contrast with central arterial stiffness, peripheral arterial stiffness is difficult to change with aerobic exercise training. PMID- 16248932 TI - Terror threat perception and its consequences in contemporary Britain. AB - The terrorist attacks of 9/11, and subsequent terrorist acts around the world, have alerted social psychologists to the need to examine the antecedents and consequences of terrorist threat perception. In these two studies, we examined the predictive power of demographic factors (age, gender, location), individual values and normative influences on threat perception and the consequences of this perception for behavioural change and close relationships. In Study 1 (N = 100), gender, benevolence values and normative influences were all correlates of threat perception, whilst sense of personal threat was correlated with increased contact with friends and family. In Study 2 (N = 240) age, gender, location, and the values of openness to change and hedonism, all predicted threat perception, which, in turn, predicted behavioural change and relationship contact. Such findings point to the important role social psychologists should play in understanding responses to these new terrorist threats. PMID- 16248933 TI - Face processing and familiarity: evidence from eye-movement data. AB - How is information extracted from familiar and unfamiliar faces? Three experiments, in which eye-movement measures were used, examined whether there was differential sampling of the internal face region according to familiarity. Experiment 1 used a face familiarity task and found that whilst the majority of fixations fell within the internal region, there were no differences in the sampling of this region according to familiarity. Experiment 2 replicated these findings, using a standard recognition memory paradigm. The third experiment employed a matching task, and once again found that the majority of fixations fell within the internal region. Additionally, this experiment found that there was more sampling of the internal region when faces were familiar compared with when they were unfamiliar. The use of eye fixation measures affirms the importance of internal facial features in the recognition of familiar faces compared with unfamiliar faces, but only when viewers compare pairs of faces. PMID- 16248934 TI - The proximity heuristic in judgments of accident probabilities. AB - In daily life, probabilities are often assessed informally through the perceived distance to a target event. Accident probabilities are believed to be high when a disastrous outcome appears to be close. This proximity heuristic can lead to exaggerated p(death) estimates in risky situations (Experiment 2), and sometimes higher probabilities for death after near-accidents than after actual accidents (Experiment 3 and 4). Near-accidents are also believed to be equally good, or better, predictors of future disasters than actual accidents, despite being less serious and more frequent (Experiment 5). The proximity heuristic influences the way people talk about their own and other people's chances (Experiment 1), and how they prepare to avoid (or, if necessary, to produce) a disastrous outcome (Experiment 6). PMID- 16248935 TI - Dissociational body experiences: differences between respondents with and without prior out-of-body-experiences. AB - Many people report having had an experience in which they felt as if their phenomenal self was separated in Cartesian space from their physical body. This phenomenon is often referred to as an 'out-of-body' (OBE) experience. Prior work has found OBE experients to score higher on measures of dissociation and to differ in regards to the perceptual experience of their body. Based upon this work, we theorized that the daily bodily experiences of people with and without a prior OBE would differ along a number of dimensions. In order to test this theory a questionnaire study was conducted. Of 243 respondents, 62 reported at least one prior OBE. Six scales on different aspects of bodily experience were administered. Respondents reporting a previous OBE were found to score significantly higher on measures of somatoform dissociation, self-consciousness, body dissatisfaction, and lower on a measure of confidence in their physical self presentation than respondents without a previous OBE. The findings are discussed as supporting a dissociational theory of the OBE. PMID- 16248936 TI - Affective valence and the self-reference effect: influence of retrieval conditions. AB - Positive trait information is typically better recalled than negative trait information when encoded in reference to the self, but not when encoded in reference to someone else or when processed for general meaning. This study examined whether this influence of affective meaning is modulated by retrieval conditions. Participants encoded positive and negative trait adjectives in reference to themselves or to a celebrity. They were then presented with either a free-recall task (Experiment 1) or a recognition memory task (Experiment 2). Positive adjectives were better recalled than negative adjectives, but only when they were encoded in reference to the self. In contrast, encoding condition and valence did not interact in the recognition memory task. Taken together, these findings suggest that the difference in memory between positive and negative self referent information is due, at least in part, to a control exerted on memory retrieval. PMID- 16248937 TI - Compulsive buying--a growing concern? An examination of gender, age, and endorsement of materialistic values as predictors. AB - Compulsive buying is an understudied, but growing, dysfunctional consumer behaviour with harmful psychological and financial consequences. Clinical perspectives treat it as a psychiatric disorder, whereas recent proposals emphasize the increasing endorsement of materialistic values as a cause of uncontrolled buying (e.g. Dittmar, 2004b; Kasser & Kanner, 2004). The present research aims to improve understanding of compulsive buying through examining gender, age, and endorsement of materialistic values as key predictors in three UK questionnaire studies, which sampled individuals who had contacted a self-help organization and residentially matched 'controls' (N = 330), consumer panelists from a multinational corporation (N = 250), and 16- to 18-year-old adolescents (N = 195). The results confirmed previously documented gender differences, and showed that younger people are more prone to compulsive buying. The central findings were that materialistic value endorsement emerged as the strongest predictor of individuals' compulsive buying, and that it significantly mediated the observed age differences. PMID- 16248938 TI - The link between composition and balance in masterworks vs. paintings of lower artistic quality. AB - In painting, composition is commonly defined as the two-dimensional arrangement of elements within the canvas. Each element is considered to have a perceptual weight. The arrangement of these weighted elements determines how balanced a painting is. It has been suggested that due to superior composition, masterworks may be more balanced than works of lower artistic quality. We tested this hypothesis by instructing our participants to rate masterworks and selections of lower artistic quality on balance. This hypothesis was not supported. Second, it has been suggested that rearranging elements within a painting may have a more detrimental effect on composition (and by extension balance) in masterworks than in selections of lower artistic quality. This view associates works of higher artistic quality with visual rightness, thereby predicting that compositional change would be more likely to cause deviations from a visually right state in masterworks. We tested this hypothesis by displacing an element within each painting to a different location, and measuring the effect on balance. In accordance with recent findings in the literature, we also took into account the severity of the compositional alterations. The results demonstrated that compositional alteration affected balance ratings equally across masterworks and selections of lower artistic quality. These results demonstrate that, although balance is a function of compositional structure, balance on its own is not sufficient to distinguish between works of varying artistic quality. To the extent that balance is considered a function of composition, the results suggest that masterworks are distinguished from works of lower artistic quality for reasons other than solely composition. PMID- 16248939 TI - Sex differences in means and variability on the progressive matrices in university students: a meta-analysis. AB - A meta-analysis is presented of 22 studies of sex differences in university students of means and variances on the Progressive Matrices. The results disconfirm the frequent assertion that there is no sex difference in the mean but that males have greater variability. To the contrary, the results showed that males obtained a higher mean than females by between .22d and .33d, the equivalent of 3.3 and 5.0 IQ conventional points, respectively. In the 8 studies of the SPM for which standard deviations were available, females showed significantly greater variability (F(882,656) = 1.20, p < .02), whilst in the 10 studies of the APM there was no significant difference in variability (F(3344,5660) = 1.00, p > .05). PMID- 16248940 TI - [Interventional therapy for hepatocellular carcinoma]. PMID- 16248941 TI - [Distribution and clinical significance of hepatitis B virus (HBV) genotypes and subtypes in HBV-infected patients]. AB - OBJECTIVE: To study hepatitis B virus (HBV) genotype and subtype distribution and its clinical significance in HBV-infected patients. METHODS: We used type/subtype specific primers and PCR to detect HBV genotype and subtype of 445 HBV-infected patients from Beijing, Changchun, Hanchuan Shenzhen, Qingyuan and Nanjing, including 7 acute hepatitis (AH), 36 asymptomatic HBV carriers (ASC), 352 chronic hepatitis (CH), 28 liver cirrhosis (LC), and 22 hepatocellular carcinoma (HCC) cases. Genotyping results were confirmed by PCR product sequencing. RESULTS: Among 445 HBV-infected patients, the proportions of genotype B, C, and B/C were 32.6% (145/445), 53.7% (239/445), and 13.7% (61/445), respectively. In genotype C, 13 (5.4%) were subtype C1, 135 (56.5%) were subtype C2, and the remaining 91 (38.1%) were neither C1 nor C2. In genotype B, 100 (69.0%) were subtype Ba, 25 (17.2%) subtype Bj, and the other 20 (13.8%) were neither Ba nor Bj. In genotype B/C, 15 (24.6%) were Ba/C2, 8 (13.1%) Bj/C2, 6 (9.8%) Ba/C1, 3 (4.9%) Bj/C1, 11 (18.0%) Ba/neither C1 nor C2, 7 (11.5%) Bj/neither C1 nor C2, and 6 (9.8%) neither Ba nor Bj/neither C1 nor C2, 2 (3.3%) neither Ba nor Bj/C1, 3 (4.9%) neither Ba nor Bj/C2. The HBV genotype and subtype distribution we found exhibited significant differences in the various clinical types of HBV infection tested, and showed that genotype C was predominant among patients with liver cirrhosis (78.6%) and hepatocellular carcinoma (86.4%) while genotype B was predominant in asymptomatic carriers (72.2%). In addition, genotype and subtype distribution showed no significant differences between male and female patients, but genotype and subtype distribution showed significant differences in patients positive or negative with HBeAg. CONCLUSION: Subtypes Ba and C2 are predominant in patients with hepatitis B from these 6 cities, and genotype C may be associated with the development of liver cirrhosis and hepatocellular carcinoma. PMID- 16248942 TI - [Analysis of prognostic factors of and to establish a predictive model for patients with chronic severe hepatitis]. AB - OBJECTIVE: To analyze prognostic factors of and to develop a prognostic model for patients with chronic severe hepatitis (CSH). METHODS: From December 1998 to October 2003, 385 in-patients being treated for chronic severe hepatitis were evaluated. The main clinical and laboratory variables were analyzed as predictive factors of survival with Cox univariate and multivariate regression models. RESULTS: The median survival time of this study group was 47 days. The survival rates at 1, 3, 6 months, 1-year, and 3 years were 66.2%, 32.9%, 26.9%, 22.9% and 17.7% respectively. Four prognostic factors were extracted using Cox's proportional hazard model; the prognostic index (PI) was calculated using the following formula consisting of these factors. PI = 0.016loge + 1.148 hepatic encephalopathy + 0.294loge (bilirubin micromol/L) - 0.826loge (prothrombin time activity). The model accurately predicted 3 months survival in an independent series of 84 patients with chronic severe hepatitis. CONCLUSION: The developed model is valuable in prognostic evaluation of chronic severe hepatitis and it may be useful to guide clinicians in selecting treatment methods for CSH. PMID- 16248943 TI - [Complete HBV DNA clone and sequence from serum samples of severe hepatitis B patients]. AB - OBJECTIVE: To study the association between hepatitis B virus (HBV) mutants and the pathogenesis of severe hepatitis B by full-length HBV genome. METHODS: Serum samples from 10 severe hepatitis B patients were collected in our hospital. Serum HBV DNAs were extracted using DNA mini Kit, and amplified by LA Taq DNA polymerase to yield full-length HBV DNA. PCR products were isolated and cloned into vector pUCm-T, then transfected into DH-5 alpha cells. Positive clones were selected and checked by digestion, and full-length HBV DNAs were sequenced. RESULTS: 4 cases were cloned into vector pUCm-T successfully and completed the full-length sequencing. Among them, 3 cases had a G to A mutation at nucleotide 1896 in pre-C region and 1 had a double mutation of T1762-A1764 in the core promoter region. Some amino acid changes occurred within the known CTL, B or T cell epitopes of the PrS2 and C regions. CONCLUSIONS: This method could serve to study the relationship between HBV genome and the pathogenesis of severe hepatitis B. PMID- 16248944 TI - [Screening and cloning of hepatitis C virus non-structural protein 4A interacting protein gene in hepatocytes]. AB - OBJECTIVE: To investigate biological functions of hepatitis C virus (HCV) non structural protein 4A (NS4A). METHODS: Yeast-two hybrid technique was performed to seek proteins in hepatocytes interacting with HCV NS4A. HCV NS4A bait plasmid was constructed by ligating the NS4A gene with carrier plasmid pGBKT7, then it was transformed into yeast AH109 (alpha type). The transformed yeast cells were amplified and mated with yeast cells Y187 (alpha type) containing liver cDNA library plasmid pACT2 in 2 x YPDA medium. Diploid yeast cells were plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) and synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing X-alpha-gal for selection two times. After extracting plasmid from blue colonies, plasmid DNA was transformed into competent E.coli and analyzed by DNA sequencing and bioinformatics methods. RESULTS: Among twenty-two positive colonies there were eleven positive for metallothionein 2A, three for eukaryotic translation elongation factor 1 alpha 1, two for albumin, two for RNA binding motif protein 21, two for myomesin, one for cytochrome C oxidase II, and one for ATPase. CONCLUSIONS: Genes of HCV NS4A interacting proteins in hepatocytes were successfully cloned and the results pave the way for studying the biological functions of NS4A and associated proteins. PMID- 16248945 TI - [Interferon-alpha and ribavirin combination therapy for co-infection of hepatitis C virus and human immunodeficiency virus]. AB - OBJECTIVE: To evaluate the clinical effect and side-effect of interferon-alpha (IFN-a) and ribavirin (RBV) combination therapy for Chinese patients with co infection of hepatitis C virus (HCV) and human immunodeficiency virus (HIV), and to compare them with only HIV infection patients. METHODS: 10 patients with HCV HIV and 17 patients with only HCV infection received 5 million units of IFNalpha 2b every other day intramuscularly, and 300 mg RBV orally three times a day. Dynamic observations were done for HCV RNA and HIV RNA loads, CD4+ and CD8+ T lymphocyte counts, liver function and blood cell measures, and the side-effects of the medicines. RESULTS: After 12 weeks and 24 weeks of IFNalpha and RBV combination therapy, mean HCV RNA levels reduced 1.14 log (t = 3.843, P < 0.01) and 2.08 log (t =6.564, P < 0.01) from the baseline at week 0 in the HCV-HIV co infection group, and reduced 1.48 log (t = 6.438, P less than 0.01) and 2.33 log (t = 7.343, P < 0.01) in the HCV infection group. Meanwhile, the HIV RNA levels decreased 1.22 log (t = 3.662, P < 0.01) and 1.73 log (t = 6.119, P < 0.01) from the base line. However, there were no obvious different changes among T lymphocyte counts of HCV-HIV and HCV patients at week 0, week 12 and week 24. All 27 patients showed satisfactory biochemical response to therapy. There were some mild or moderate influenza-like symptoms, intestinal discomfort and decreased blood cell counts in the early stages of the treatments. No neuropsychic and auto immune disorders were found. CONCLUSIONS: IFNalpha-2b and RBV combination therapy showed similar anti-HCV effects during the 24 week treatment for HCV-HIV and HCV infected patients, and some anti-HIV effect was also observed. No obvious different biochemical responses and side-effects were found between the above two groups. PMID- 16248946 TI - [A study of 10-23 DNAzyme inhibiting the expression of hepatitis B virus genes]. AB - OBJECTIVES: To explore, on the cell level, the possibility of using 10-23DNAzyme, as a new genetherapy in treating hepatitis B. METHODS: Phosthorothioate 10-23DRz (DRz-S) and 10-23DRz specific to HBV pre-C/C gene ORFA2031 were designed and synthesized, and the inhibition effects of 10-23DRz-S and 10-23DRz on the expression of HBV gene in HepG2 2.2.15 cells were observed. RESULTS: The expression of HBV gene was remarkably depressed after 2.2.15 cells were transfected by DRz-S and DRz. The maximum inhibition was 93.75% and 90.26%. The inhibition effect was maintained for 96 hours, and the inhibition time of DRz-S was longer than that of DRz. The maximum inhibition of DRz-S was lower than that of DRz. The efficiency of inhibiting HBsAg and HBeAg in 2.2.15 cells transfected by DRz-S and DRz was higher than that by antisense oligonucleotides for the same target genes. It had no remarkable effect on the replication of HBV DNA and no toxicity to the 2.2.15 cells. CONCLUSION: 10-23DRz can highly block the expression of HBV gene in the 2.2.15 cell model and it can serve as a specific and effective anti-HBV gene therapeutic means. PMID- 16248947 TI - [A study of the down-regulation effect of hepatitis B virus pre-protein S2 on inducible nitric oxide synthase gene promoter]. AB - OBJECTIVES: To investigate the regulating effect of HBV pre-S2 protein on iNOS gene promoter and the molecular biological mechanisms of pre-S2 protein in HBV pathogenicity. METHODS: Polymerase chain reaction (PCR) technique was employed to amplify the sequence of iNOS promoter and 3 deletion mutants using HepG2 genomic DNA as the template, and the products were cloned into the pGEM-T vector. The iNOS gene and 3 deletion mutants were cut from T- iNOS by Kpn I and Xho I, and then cloned into pCAT3-Basic. The resulting vectors were named p1-iNOSp, p2 iNOSp, p3-iNOSp, and p4-iNOSp. Each of the reporter vectors was transfected into the HepG2 cell line and cotransfected into HepG2 cells with pcDNA3.1(-)-pre-S2 by FuGENE 6 transfection reagents. The HepG2 cells transfected with pCAT3-Basic were used as a negative control. The activity of CAT in HepG2 cells transfected was detected by an ELISA kit 48 hours after the transfection, which reflected the regulating effect of HBV pre-S2 protein on iNOS gene promoter activity. RESULTS: The expressive vector pcDNA3.1(-)-pre-S2 and report vector pCAT3-iNOSp were constructed and confirmed by restriction enzyme digestion and sequencing. The expression of pcDNA3.1(-)-pre-S2 in HepG2 cells could down-regulate the activity of p1-iNOSp, p3-iNOSp, and the inhibition rate was 54.7% and 79.5%, respectively. The expression of pcDNA3.1(-)-pre-S2 in HepG2 cells had no regulatory effects on p2-iNOSp and p4-iNOSp. CONCLUSION: It is suggested that HBV pre-S2 protein can down-regulate iNOS gene promoter. PMID- 16248948 TI - [A comparative study of MRI manifestations and pathological changes in hepatocellular carcinoma treated by transcatheter arterial chemoembolization with lipiodol]. AB - OBJECTIVES: To analyze the MRI manifestations and pathological changes of hepatocellular carcinoma (HCC) after transcatheter arterial chemoembolization (TACE) with lipiodol. METHODS: 23 patients with 31 HCC lesions treated by TACE underwent MRI examination within 1 week before their surgical resections. MRI was performed with SE sequence (T1WI and FSE T2WI) and FMPSPGR sequence dynamic multi phase contrast scans. All resected specimens were cut into 5-10mm thick slices, corresponding to the same plane as that of MRI scans. The specimens were wholly embedded in paraffin, serial sections made and stained with hematoxylin and eosin. The MRI findings were thus compared with the pathology of the specimen sections. RESULTS: (1) MRI findings: In all 31 lesions, the signal intensity of lesions varied and was mostly heterogeneous on SE T1WI and T2WI images. Three lesions were inhomogeneous hyper-intensity and the other 28 lesions were iso- or hypo-intensity on FMPSPGR plain scannings. Twenty-two lesions were enhanced on early-phase dynamic scanning, and no enhancement was found in the other 9 lesions. Partial enhancement was also seen in 6 lesions on delay-phase dynamic scanning. (2) Pathologically, no coagulation necrosis was found in 2 specimens, but 6 lesions showed complete coagulation necrosis and 23 showed various degrees of it. The other pathological changes found included intra-tumoral hemorrhage (n=10), intra-lesional fibrotic septa formation (n=5), capsule-like fibrotic tissue proliferation around the lesions (n=12), inflammatory infiltration (n=28), focal mucoid degeneration (n=2), focal hyaline degeneration (n=2), and lipiodol retention (n=6). (3) Radiological-pathological correlation study: hyper-intense areas on T1WI corresponded to areas of coagulation necrosis with or without hemorrhage and of residual viable tumor; iso- and hypo-intense corresponded to areas of coagulation necrosis or residual viable tumor. Hyper-intense areas on T2WI corresponded to those of residual viable tumor or coagulation necrosis with hemorrhage, and iso-intense areas corresponded to those of coagulation necrosis, small residual viable tumor or intra-lesional fibrotic septa formation, and hypo intense areas corresponded to those of coagulation necrosis or intra-lesional fibrotic septa formation. Areas of enhancement within the lesions on the early phase dynamic-contrast images corresponded to areas of residual viable tumors, while areas of no enhancement were those of coagulation necrosis, hemorrhage, intra-lesional fibrotic septa formation or small residual viable tumors. Areas of enhancement on the delay-phase dynamic scanning were those of residual viable tumors or intra-lesional fibrotic septa formation, while no enhancement corresponded to the areas of residual viable tumors, coagulation necrosis, and hemorrhage. Areas of enhancement on the delay-phase dynamic scanning corresponded to those areas of fibrosis tissue or residual viable tumors. Inflammatory infiltration was found in areas of different signal intensity on MRI images. CONCLUSIONS: (1) Different pathological changes in HCCs after TACE are represented by various signal intensities on SE sequence images. The only area of hypo-intensity on T2WI has a specificity in representing coagulation necrosis. (2) FMPSPGR sequence dynamic MRI is superior to SE sequence in demonstrating and determining the necrosis and residual viable tumor. Enhanced areas within the lesions on the early-phase dynamic-contrast images represent residual viable tumors and the enhancement of capsule on early-phase dynamic-contrast images also represent subcapsular residual viable tumors. (3) MRI can demonstrate accurately the areas of necrosis and residual viable HCC tissues after TACE and evaluate the effect of TACE. PMID- 16248949 TI - [Inhibition of mutant-type p53 by a chimeric U6 maxizyme in hepatocellular carcinoma cell lines]. AB - OBJECTIVE: To study the inhibition of maxizyme (Mz) directed against the mutant type p53 gene (mtp53) at codon 249 in exon 7 (AGG --> AGT) both in cell-free system and in MHCC97 cell lines. METHODS: Maxizyme and control mutant maxizyme (G5 --> A5) were designed by computer and cloned into the eukaryotic expression vector pBSKneoU6 (pU6Mz, pU6asMz). Mz was driven by T7 RNA polymerase promoter in vitro. In the cell lines, U6 promoter was driven by RNA PolIII. The mutant type p53 gene fragment was cloned into the pGEM-T vector under the T7 promoter control. The 32P-labeled mtp53 transcript was the target RNA. Cold maxizyme transcripts were incubated with 32P-labeled target RNA in vitro. pU6Mz was introduced into MHCC97 cells by Lipofectamine2000 and mtp53 expression was analyzed by RT-PCR and Western blot. RESULTS: In vitro cleavage showed that pU6Mz was very active with cleavage efficiency of 42% while pU6asMz was not. The wild type p53 was not cleaved. Partial down-regulation of mtp53 mRNA and mtp53 protein were observed in MHCC97 cells transfected with pU6Mz but not those with pU6asMz. The proliferation of MHCC cells was inhibited by MTT analysis. CONCLUSION: Our findings suggest that the chimeric U6 maxizyme against the mtp53 is a new promising gene therapeutic agent in treating hepatocellular carcinoma. PMID- 16248950 TI - [A cDNA microarray study of the differential expression of genes in signal transduction pathway during hepatocarcinogenesis in tree shrews]. AB - OBJECTIVES: To study the differential expression of genes in signal transduction pathway (STP) during the hepatocarcinogenesis in tree shrews induced by AFB1 and/or HBV and to elucidate the molecular mechanism of hepatocellular carcinoma (HCC) development. METHODS: Adult tree shrews were divided into three groups: Group A was fed AFB1 only, Group B was infected firstly with HBV then fed AFB1 as in Group A, Group C served as the normal control. Liver biopsies were obtained at the 30th, 60th and 90th week of the experiment or until HCC occurred and the animals were sacrificed. Tree shrew-specific cDNA microarray was applied for detecting the differential expression of corresponding genes in each group at different time points during the experiment, and real time RT PCR was applied to verify the results of the cDNA microarray. RESULTS: Genes of IGF-II, C-rel, and NF-kappaB2 were differentially expressed between para-cancerous tissues and HCC tissues in both group A and group B, and the differential expression of bcl-2, cyclin A and CNTF was only seen in group B. Between the experimental groups A and B and the control group C, there were differential expressions of CNTF and cyclin A in the early 30th week and middle 60th week stage of hepatocarcinogenesis in tree shrews. Real time RT PCR results showed that the expression level of IGF-II and C-Rel in group A and of IGF-II in group B in HCC tissues were significantly lower than that in the adjacent non-cancerous tissues and in the biopsies taken at the 30th and 60th week of the experiment. Nevertheless, there were no significant differences between the para-cancerous tissues and the cancer tissues at the 30th and 60th week. These results were consistent with the cDNA microarray assay. The expression levels of C-Rel and CNTF in group B were not obviously altered in the para-cancerous tissues, HCC and at the 60th week, but they were significantly lower in these tissues than that in the tissues at the 30th week. In group A, the expression levels of CNTF in adjacent liver and HCC tissues were higher than that in para-cancerous lesions, but the difference did not reach a statistically significant level. In group C, the expression level of IGF-II, C Rel and CNTF at different stages showed no significant differences, which was consistent with the cDNA microarray results. CONCLUSIONS: To apply the tree shrew specific cDNA microarray to detect the differential expression of genes related to signal transduction pathway during tree shrew hepatocarcinogenesis could be a valuable utility for further comprehending the mechanism of HCC. IGF-II, NF kappaB2, C-rel, Bcl-2, and cyclin A. CNTF may be involved in the occurrence and progress of HCC in tree shrews. PMID- 16248951 TI - [Inhibitory effect of cyclin D1 antisense cDNA on human hepatocarcinoma cell line HepG2]. AB - OBJECTIVE: To investigate the effect of antisense cDNA of cyclin D1 on the cyclin D1 gene expression and cell proliferation of human hepatocarcinoma HepG2 cells in vitro. METHODS: Plasmids containing cyclin D1 antisense cDNA were constructed and transfected into HepG2 cells. Their effects on cell proliferation were examined by MTT method, RT-PCR, immunohistochemical means, and flow cytometry. RESULTS: Cyclin D1 antisense cDNA significantly inhibited the growth of HepG2 cells. The inhibition peaked at 48 hour after transfection by MTT method. RT-PCR analysis showed that cyclin D1 antisense cDNA down-regulated cyclin D1 at the mRNA levels. Expression level of cyclin D1 protein was also decreased as shown by immunohistochemical studies. Cell-cycle analysis by flow cytometry showed that transfected HepG2 cells were arrested at the G1 phase of the cell cycle. CONCLUSIONS: Our data suggest that cyclin D1 antisense cDNA could specifically inhibit the expression of cyclin D1 mRNA and protein and regulate cell cycle and cell proliferation of HepG2 cells. Cyclin D1 antisense cDNA may serve as a potential antitumor strategy in regulating cell-cyclin treating advanced HCCs. PMID- 16248952 TI - [Use of autologous tumor vaccine in preventing recurrence of hepatocellular carcinoma]. AB - OBJECTIVE: To evaluate the effects of autologous tumor vaccines in preventing recurrences of hepatocellular carcinoma (HCC). METHODS: From March 1999 to June 2003, 80 patients with HCC undergoing resections were randomly assigned into a tumor vaccine group (n=40) and a control group (n=40). Tumor vaccines, consisting of formalin-fixed HCC tissue fragments, biodegradable sustained-releasers of granulocyte-macrophage-colony stimulating factor, interleukin-2, and an adjuvant, were developed. Every vaccine group patient received 3 vaccinations at a 2-week interval and the control group just received the adjuvant. Delayed-type hypersensitivity (DTH) test and recurrent rates were analyzed. RESULTS: Eight patients of the vaccine group and five patients of the control group were lost in the follow-up. Thirty-two patients completed the tumor vaccine procedure and no essential adverse effects occurred. 23/32 patients developed DTH responses against the fragments of HCC. The follow-up averaged 34.3 months (from 15 to 55 months). 1-, 2-, 3-year recurrence rates of the vaccine group were 12.6%, 35.9% and 54.0%, respectively; 1-, 2-, 3-year recurrence rates of the control group were 31.6%, 61.3% and 72.1%, respectively. The recurrent rate was significantly better in the tumor vaccine group than in the control group (P = 0.037). CONCLUSIONS: Autologous tumor vaccine is a promising adjunctive modality to prevent recurrence of human HCC. PMID- 16248953 TI - [The expression and the significance of L-FABP and FATP4 in the development of nonalcoholic fatty liver disease in rats]. AB - OBJECTIVE: To study the effect of liver fatty acid binding protein(L-FABP) and fatty acid transport protein (FATP4) in the development of nonalcoholic fatty liver disease (NAFLD) in rats. METHODS: The expression of L-FABP and FATP4 genes was examined in fatty liver rats by reverse transcription and polymerase chain reaction amplification and Western blot methods. RESULTS: In the high fat diet group (F), mRNA and protein expression of L-FABP and FATP4 were increased at 2 weeks, and they increased remarkably at 12 weeks (P < 0.05; L-FABP mRNA F=124.9, protein expression F=92.6; FATP4 mRNA F=602.9, protein expression F=108.8). CONCLUSION: The high expression of L-FABP and FATP4 at the early stage is an adaptive reaction of the body, With the advanced expression of the L-FABP and FATP4, it can lead to a fatty acid disequilibrium and then result in nonalcoholic fatty liver disease in the rats. PMID- 16248954 TI - [Experimental study on the pathological effect of Helicobacter pylori on liver tissues]. AB - OBJECTIVE: To observe whether H. pylori inoculated by oral route could arrive in livers and cause liver inflammation as an independent etiological factor. METHODS: C57BL/6 mice were orally inoculated with H. pylori SS1 strains and fed for 8 months. H. pylori colonization and pathologic consequences were studied in the liver and gallbladder tissues of the mice; the blood, liver tissue and gastric mucosa were obtained and cultured for H. pylori growth; The bacterial DNA extracted from the liver, bile and blood was examined by nested PCR for H. pylori genes. 16S rRNA PCR amplicons were sequenced and compared with the sequencing results of 16S rRNA PCR amplicons of the bacteria cultured from gastric mucosa and the inoculated H. pylori SS1. RESULTS: The bacterial DNA extracted from the liver, bile and blood of the infected mice was detected for H. pylori genes by nested PCR. Six of the 15 samples were positive (40%) in the liver, 6 of 10 samples in the bile (60%), and 2 of 10 samples in the blood (20%). Sequencing results of 16S rRNA PCR products of the livers showed 100% homogeneity when compared with the cultured H. pylori from gastric mucosa and inoculated H. pylori SS1. H. pylori was found in 4 liver tissues of the 15 infected mice (26.7%) and 6 in the gallbladders (40%). Infiltrations of lymphocyte cells along hepatic sinusoids and a lower degree infiltration around interlobular arteries and veins were observed; ballooning degeneration was also observed in some hepatocytes. CONCLUSION: H. pylori inoculated by oral route could arrive in the liver and cause inflammation as an independent etiological factor. The routes which the microorganisms took to reach the livers may involve hematogenous and/or biliary system dissemination. PMID- 16248955 TI - [The principle of interventional therapy of hepatoma]. PMID- 16248956 TI - [Approaches for interventional therapy of hepatoma]. PMID- 16248958 TI - [Effects of serum with drug "KXEH" on activated hepatic stellate cells]. PMID- 16248957 TI - [Therapeutic effect of diammonium glycyrrhizinate lipid ligand on carbon tetrarchloride induced fatty livers in rats]. PMID- 16248959 TI - [The protective effect of MG132 on post-ischemia reperfusion injury of rat livers by activation of nuclear factor kappa B]. PMID- 16248960 TI - [An in vitro study on the mutant of HBV treated with interferon]. PMID- 16248961 TI - [Clinical significance of the dysregulated expression of von Willebrand factor cleaving protease in patients with primary hepatocarcinoma]. PMID- 16248962 TI - [Development on HLA-restricted HBV-specific CTL epitopes]. PMID- 16248963 TI - Multifunctional role of Fas-associated death domain protein in apoptosis. PMID- 16248964 TI - Rescuing developing thymocytes from death by neglect. AB - The major function of the thymus is to eliminate developing thymocytes that are potentially useless or autoreactive, and select only those that bear functional T cell antigen receptors (TCRs) through fastidious screening. It is believed that glucocorticoids (GCs) are at least in part responsible for cell death during death by neglect. In this review, we will mainly cover the topic of the GC induced apoptosis of developing thymocytes. We will also discuss how thymocytes that are fated to die by GCs can be rescued from GC-induced apoptosis in response to a variety of signals with antagonizing properties for GC receptor (GR) signaling. Currently, a lot of evidence supports the notion that the decision is made as a result of the integration of the multiple signal transduction networks that are triggered by GR, TCR, and Notch. A few candidate molecules at the converging point of these multiple signaling pathyways will be discussed. We will particularly describe the role of the SRG3 protein as a potent modulator of GC induced apoptosis in the crosstalk. PMID- 16248965 TI - Caspase recruitment domain (CARD) as a bi-functional switch of caspase regulation and NF-kappaB signals. PMID- 16248967 TI - Nuclear factor-kappaB activation: a question of life or death. AB - Apoptosis is a mode of cell death that plays an important role in both pathological and physiological processes. Research during the last decade has delineated the entire machinery needed for cell death, and its constituents were found to pre-exist in cells. The apoptotic cascade is triggered when cells are exposed to an apoptotic stimulus. It has been known for several years that inhibitors of protein synthesis can potentiate apoptosis that is induced by cytokines and other inducers. Until 1996, it was not understood why protein synthesis inhibitors potentiate apoptosis. Then three reports appeared that suggested the role of the transcription factor NF-kappaB activation in protecting the cells from TNF-induced apoptosis. Since then several proteins have been identified that are regulated by NF-kappaB and are involved in cell survival, proliferation, and protection from apoptosis. It now seems that when a cell is attacked by an apoptotic stimulus, the cell responds first by activating anti apoptotic mechanisms, which may or may not be followed by apoptosis. Whether or not a cell undergoes proliferation, the survival, or apoptosis, appears to involve a balance between the two mechanisms. Inhibitors of protein synthesis seem to suppress the appearance of protein that are involved in anti-apoptosis. The present review discusses how NF-kappaB controls apoptosis. PMID- 16248966 TI - Apoptotic signaling pathways: caspases and stress-activated protein kinases. AB - Apoptotic cell death is an active process mediated by various signaling pathways, which include the caspase cascade and the stress-activated protein kinase pathways. The caspase cascade is activated by two distinct routes: one from cell surface and the other from mitochondria. Activation of the route from cell surface requires the cellular components that include membrane receptors, adaptor proteins such as TRADD and FADD, and caspase-8, while activation of the other from mitochondria requires Apaf-1, caspase-9, and cytosolic cytochrome c. On the other hand, persistent stimulation of the stress-activated protein kinase pathway is also shown to mediate apoptosis in many cell types. Gene-targeting studies with jnk- or jip-null mice, in particular, strongly suggest that this signaling pathway plays a pivotal role in the cellular machinery for apoptosis. PMID- 16248968 TI - Potassium efflux during apoptosis. PMID- 16248969 TI - TGF-beta-dependent cell growth arrest and apoptosis. PMID- 16248970 TI - Cytokine synergism in apoptosis: its role in diabetes and cancer. AB - The effects of individual cytokine on apoptosis have been extensively studied. However, the effect of the cytokine combination, or the synergistic effect of cytokines on cell death, has not been widely studied, though synergism between cytokines has been documented in a variety of biological situations. In our effort to identify the final death effector molecule(s) in autoimmune diabetes, we inadvertently became interested in the cytokine synergism. We discovered that IFNgamma/TNFalpha synergism, rather than the Fas ligand as currently believed, is responsible for the apoptosis of pancreatic islet cells both in vitro and in vivo. We also studied similar cytokine synergism in cancer cell deaths, and noted the similarities and dissimilarities between cancer cell death and islet cell death. PMID- 16248971 TI - Modulation of life and death by the tumor necrosis factor receptor-associated factors (TRAFs). AB - The TNF receptor-associated factor (TRAF) family is a group of adapter proteins that link a wide variety of cell surface receptors. Including the TNF and IL-1 receptor superfamily to diverse signaling cascades, which lead to the activation of NF-kappaB and mitogen-activated protein kinases. In addition, TRAFs interact with a variety of proteins that regulate receptor-induced cell death or survival. Thus, TRAF-mediated signals may directly induce cell survival or interfere with the death receptor-induced apoptosis. PMID- 16248972 TI - Cellular and molecular pathways of ischemic neuronal death. AB - Three routes have been identified triggering neuronal death under physiological and pathological conditions. Excess activation of ionotropic glutamate receptors cause influx and accumulation of Ca2+ and Na+ that result in rapid swelling and subsequent neuronal death within a few hours. The second route is caused by oxidative stress due to accumulation of reactive oxygen and nitrogen species. Apoptosis or programmed cell death that often occurs during developmental process has been coined as additional route to pathological neuronal death in the mature nervous system. Evidence is being accumulated that excitotoxicity, oxidative stress, and apoptosis propagate through distinctive and mutually exclusive signal transduction pathway and contribute to neuronal loss following hypoxic-ischemic brain injury. Thus, the therapeutic intervention of hypoxic-ischemic neuronal injury should be aimed to prevent excitotoxicity, oxidative stress, and apoptosis in a concerted way. PMID- 16248973 TI - Effect of neurotrophic factors on neuronal stem cell death. AB - Neural cell survival is an essential concern in the aging brain and many diseases of the central nervous system. Neural transplantation of the stem cells are already applied to clinical trials for many degenerative neurological diseases, including Huntington?'s disease, Parkinson?'s disease, and strokes. A critical problem of the neural transplantation is how to reduce their apoptosis and improve cell survival. Neurotrophic factors generally contribute as extrinsic cues to promote cell survival of specific neurons in the developing mammalian brains, but the survival factor for neural stem cell is poorly defined. To understand the mechanism controlling stem cell death and improve cell survival of the transplanted stem cells, we investigated the effect of plausible neurotrophic factors on stem cell survival. The neural stem cell, HiB5, when treated with PDGF prior to transplantation, survived better than cells without PDGF. The resulting survival rate was two fold for four weeks and up to three fold for twelve weeks. When transplanted into dorsal hippocampus, they migrated along hippocampal alveus and integrated into pyramidal cell layers and dentate granule cell layers in an inside out sequence, which is perhaps the endogenous pathway that is similar to that in embryonic neurogenesis. Promotion of the long term-survival and differentiation of the transplanted neural precursors by PDGF may facilitate regeneration in the aging adult brain and probably in the injury sites of the brain. PMID- 16248974 TI - Apoptotic cell death following traumatic injury to the central nervous system. AB - Apoptotic cell death is a fundamental and highly regulated biological process in which a cell is instructed to actively participate in its own demise. This process of cellular suicide is activated by developmental and environmental cues and normally plays an essential role in eliminating superfluous, damaged, and senescent cells of many tissue types. In recent years, a number of experimental studies have provided evidence of widespread neuronal and glial apoptosis following injury to the central nervous system (CNS). These studies indicate that injury-induced apoptosis can be detected from hours to days following injury and may contribute to neurological dysfunction. Given these findings, understanding the biochemical signaling events controlling apoptosis is a first step towards developing therapeutic agents that target this cell death process. This review will focus on molecular cell death pathways that are responsible for generating the apoptotic phenotype. It will also summarize what is currently known about the apoptotic signals that are activated in the injured CNS, and what potential strategies might be pursued to reduce this cell death process as a means to promote functional recovery. PMID- 16248975 TI - Akt: versatile mediator of cell survival and beyond. AB - The serine/threonine kinase Akt has been intensely studied for its role in growth factor-mediated cell survival for the past 5 years. On the other hand, the ongoing research effort has recently uncovered novel regulatory mechanisms and downstream effectors of Akt that demonstrate the involvement of Akt in other cellular functions such as cell cycle progression, angiogenesis, and cancer cell invasion/metastasis. Furthermore, recent studies using whole model organisms suggest additional roles for Akt in important diseases such as aging and diabetes. The following review addresses these recent advances in the understanding of Akt function. PMID- 16248976 TI - Nitric oxide as a pro-apoptotic as well as anti-apoptotic modulator. AB - Nitric oxide (NO), synthesized from L-arginine by NO synthases, is a small, lipophilic, diffusible, highly reactive molecule with dichotomous regulatory roles in many biological events under physiological and pathological conditions. NO can promote apoptosis (pro-apoptosis) in some cells, whereas it inhibits apoptosis (anti-apoptosis) in other cells. This complexity is a consequence of the rate of NO production and the interaction with biological molecules such as metal ion, thiol, protein tyrosine, and reactive oxygen species. Long-lasting overproduction of NO acts as a pro-apoptotic modulator, activating caspase family proteases through the release of mitochondrial cytochrome c into cytosol, up regulation of the p53 expression, and alterations in the expression of apoptosis associated proteins, including the Bcl-2 family. However, low or physiological concentrations of NO prevent cells from apoptosis that is induced by the trophic factor withdrawal, Fas, TNFalpha/ActD, and LPS. The anti-apoptotic mechanism is understood on the basis of gene transcription of protective proteins. These include: heat shock protein, hemeoxygenase, or cyclooxygenase-2 and direct inhibition of the apoptotic executive effectors caspase family protease by S nitrosylation of the cysteine thiol group in their catalytic site in a cell specific way. Our current understanding of the mechanisms by which NO exerts both pro- and anti-apototic action is discussed in this review article. PMID- 16248977 TI - Regulation of apoptosis by nitrosative stress. AB - Nitrosative stress can prevent or induce apoptosis. It occurs via S-nitrosylation by the interaction of nitric oxide (NO) with the biological thiols of proteins. Cellular redox potential and non-heme iron content determine S-nitrosylation. Apoptotic cell death is inhibited by S-nitrosylation of the redox-sensitive thiol in the catalytic site of caspase family proteases, which play an essential role in the apoptotic signal cascade. Nitrosative stress can also promote apoptosis by the activation of mitochondrial apoptotic pathways, such as the release of cytochrome c, an apoptosis-inducing factor, and endonuclease G from mitochondria, as well as the suppression of NF-kB activity. In this article we reviewed the mechanisms whereby S-nitrosylation and nitrosative stress regulate the apoptotic signal cascade. PMID- 16248978 TI - Synthetic bile acids: novel mediators of apoptosis. PMID- 16248979 TI - Ras/ERK signaling pathway mediates activation of the p21WAF1 gene promoter in vascular smooth muscle cells by platelet-derived growth factor. AB - We previously demonstrated that platelet-derived growth factor (PDGF) induces the cyclin-dependent kinase inhibitor p21/WAF1 promoter in vascular smooth muscle cells (VSMC) via activation of a Sp1 site in VSMC. In this report, the role and relevance of the signaling pathway in the transcriptional regulation of p21WAF1 in VSMC was examined. PDGF stimulated the expression of p21WAF1 in VSMC, as evidenced by Immunoblot and Northern blot analyses. Treatment with PD98059, a specific MEK inhibitor, and the transient expression of VSMC with DN-MEK1 plasmid effectively down-regulated PDGF-induced p21WAF1 expression and promoter activity, respectively. Furthermore, the transactivation of PDGF-stimulated Sp1 was inhibited by treatment with PD98059 and the transient expression of VSMC with the DN-MEK1 plasmid. Finally, the transient transfection of VSMC with a dominant negative Ras (RasN17) suppressed PDGF-induced ERK activity, p21WAF1 expression, and promoter activity. The overexpression of RasN17 also abolished PDGF stimulated Sp1 activity. In conclusion, the findings herein presented indicate that the activation of the Ras/ERK pathway contributes to the induction of p21WAF1 expression in VSMC. In addition, the transcription factor Sp1 that is involved in the Ras/ERK-mediated control of p21WAF1 regulation in VSMC in response to PDGF has now been identified. PMID- 16248980 TI - Retinoid metabolism during development of liver cirrhosis. AB - The changes in retinoid metabolism have been documented in liver cirrhosis. However, the dynamic alterations in levels of this vitamin between circulation and liver during development of the liver cirrhosis are not well understood. The aim of this study was to measure retinoids in the liver and circulation in parallel, during and after development of cirrhosis induced by carbon tetrachloride and thioacetamide. Retinoid levels were measured by HPLC. A decrease in retinaldehyde and total retinol, together with an increase in retinoic acid was evident in liver from both carbon tetrachloride or thioacetamide treated rats within a month after initiation of treatment. Activity of enzymes involved in retinoid metabolism such as retinaldehyde oxidase, retinaldehyde dehydrogenase, and retinaldehyde reductase were decreased in the liver. In parallel, levels of retinol and retinaldehyde in the serum were increased while retinoic acid was decreased. This study indicates that during development of cirrhosis, there is reciprocal transfer of retinoid metabolites between the circulation and the liver. PMID- 16248981 TI - In vitro antimicrobial and resistance-modifying activities of aqueous crude khat extracts against oral microorganisms. AB - OBJECTIVES: To assess antimicrobial activities of aqueous crude khat (Catha edulis) extracts against a panel of oral microorganisms and to test their ability to modify bacterial resistance to tetracycline and penicillin in vitro. DESIGN: Lyophilized aqueous extracts were prepared from three khat cultivars. The agar dilution method of the NCCLS was used to test the extracts, at concentrations of 20-1.25 mg/ml, against 33 oral strains. MIC was defined as the lowest concentration at which there was no visible growth. Slight growth was defined as marked growth reduction (MGR). The E-test was used to determine the MICs of tetracycline and penicillin-G for three resistant strains in absence and presence of a sub-MIC of the khat extracts (5mg/ml). RESULTS: Eighteen strains (55%) were sensitive to the extracts (MICs 5-20 mg/ml). Most of these were periodontal pathogens with Porphyromonas gingivalis and Tannerella forsythensis being the most susceptible (MIC 5-10mg/ml). Veillonella parvula, Actinomyces israelii and some streptococci were not sensitive. Except for Lactobacillus acidophilus that showed MGR at 1mg/ml, cariogenic species were neither sensitive. The extracts were active against Streptococcus pyogenes (MIC 10-20 mg/ml) but not against Candida albicans and Staphylococcus aureus. The presence of the khat extracts at a sub-MIC resulted in a 2-4-fold potentiation of the tested antibiotics against the resistant strains. CONCLUSIONS: Khat has water-soluble constituents possessing selective antibacterial activity against oral bacteria. There is preliminary evidence for presence of an antibiotic resistance-modifying component. Further investigation is needed to identify the active components and assess their clinical relevance. PMID- 16248983 TI - Evaluation of the role of sigma B in Mycobacterium smegmatis. AB - The alternate sigma factor, sigB, is known to play a crucial role in maintaining the stationary phase in mycobacteria. In this communication, we have studied the proteomics of Mycobacterium smegmatis mc(2)155 and its two derivatives, one of which has a disrupted sigB gene and the other, PMVSigB, which contains a multicopy plasmid containing sigB. We have identified by two-dimensional gel analyses, several proteins that are over-expressed in PMVSigB compared to mc(2)155. These proteins are either stress proteins or participate actively in different metabolic pathways of the organisms. On the other hand, when sigB deleted mycobacteria were grown until the stationary phase and its two dimensional protein profile was compared to that of mc(2)155, few DNA binding proteins were found to be up-regulated. We have shown recently that upon over expressing sigB, the cell surface glycopeptidolipids of M. smegmatis are hyperglycosylated, a situation similar to what was observed for nutritionally starved bacteria. Gene expression profile through quantitative PCR presented here identified a Rhamnosyltransferase responsible for this hyperglycosylation. PMID- 16248982 TI - Galectin-3 interacts with membrane lipids and penetrates the lipid bilayer. AB - The precise mechanism by which galectin-3 and other cytosolic proteins that lack signal peptides are secreted is yet to be elucidated. In the present analyses, we determined that galectin-3, a beta-galactoside binding protein, can interact directly with membrane lipids in solid phase binding assays. More interestingly, we determined by spectrophotometric methods that it can spontaneously penetrate the lipid bilayer of liposomes in either direction. These findings suggest that galectin-3 on its own has the capacity to traverse the lipid bilayer. Whereas the situation is rather simplified in liposomes, the interaction of galectin-3 with the plasma membrane may involve cholesterol-rich membrane domains where galectin 3 can be concentrated and form multimers or interact covalently with other proteins. PMID- 16248984 TI - Dietary vitamin C down-regulates inflammatory gene expression in apoE4 smokers. AB - The deleterious impact of cigarette smoking on cardiovascular health may be in part attributable to a free radical mediated proinflammatory response in circulating monocytes. In the current investigation, the impact of vitamin C supplementation on monocyte gene expression was determined in apoE4 smokers versus non-smokers. A total of 10 smokers and 11 non-smokers consumed 60mg/day of vitamin C for four weeks and a fasting blood sample was taken at baseline and post-intervention for the determination of plasma vitamin C and monocyte gene expression profiles using cDNA array and real time PCR. In apoE4 smokers, supplementation resulted in a 43% increase in plasma vitamin C concentrations. Furthermore, a number of genes were differentially expressed more than 2-fold in response to treatment, including a downregulation of the proinflammatory mediators tumor necrosis factor (TNF) beta, TNF receptor, neurotrophin-3 growth factor receptor, and monocyte chemoattractant protein 1 receptor. The study has identified a number of molecular mechanisms underlying the benefit of vitamin C supplementation in smokers. PMID- 16248985 TI - Novel progerin-interactive partner proteins hnRNP E1, EGF, Mel 18, and UBC9 interact with lamin A/C. AB - The Hutchinson-Gilford progeria syndrome (HGPS or progeria) is an apparent accelerated aging disorder of childhood. Recently, HGPS has been characterized as one of a growing group of disorders known as laminopathies, which result from genetic defects of the lamin A/C (LMNA) gene. The majority of HGPS mutant alleles involve a silent mutation, c.2063C>T resulting in G608G, that generates a cryptic splicing site in exon 11 of LMNA and consequently truncates 50 amino acids near the C-terminus of pre-lamin A/C. To explore possible mechanisms underlying the development of HGPS, we began a search for proteins that would uniquely interact with progerin (the truncated lamin A in HGPS) using a yeast two-hybrid system. Four new progerin interactive partner proteins were identified that had not been previously found to interact with lamin A/C: hnRNP E1, UBC9 (ubiquitin conjugating enzyme E2I), Mel-18, and EGF1. However, using control and progeria fibroblasts, co-immunoprecipitation studies of endogenous proteins did not show differential binding affinity compared to normal lamin A/C. Thus, we did not find evidence for uniquely interacting partner proteins using this approach, but did identify four new lamin A/C interactive partners. PMID- 16248987 TI - Novelty-induced correlation between visual neurons and the hippocampal theta rhythm in sleep and wakefulness. AB - Various rhythms have been shown to affect sensory processing such as the waking sleep cycle and the hippocampal theta waves. Changes in the firing of visual lateral geniculate nucleus neurons have been reported to be dependent on the animal's behavioral state. The lateral geniculate extracellular neuronal firing and hippocampal field activity were recorded in chronically implanted animals to analyze the relationship during quiet wakefulness and sleep associated with stimulation shifts that may introduce novelty. During wakefulness, a change in light flash stimulation pattern (stimuli frequency shift, stimuli on and off) caused an increment in the theta band power in 100% of the cases and a phase locking of the spikes in 53% of the recorded neurons. During slow wave sleep, there were no consistent changes in the theta power notwithstanding 13% of the neurons exhibited phase-locking, i.e., novelty may induce changes in the temporal correlation of visual neuronal activity with the hippocampal theta rhythm in sleep. The present results suggest that visual processing in slow wave sleep exists, while auditory information and learning were reported during slow wave sleep in animals and newborn humans. The changes in the theta power as well as in the neuronal phase-locking amount indicate that in slow wave sleep, the ability of the hippocampus to detect/process novelty, although present, may be decreased. This is consistent with the noticeable decrease in awareness of the environment during sleep. PMID- 16248986 TI - Lack of efficacy of the substance p (neurokinin1 receptor) antagonist aprepitant in the treatment of major depressive disorder. AB - BACKGROUND: An early clinical trial suggested that the substance P (neurokinin(1) receptor) antagonist, aprepitant, might provide a unique mechanism of antidepressant activity. Phase III trials were conducted to confirm these findings. METHODS: Five 8-week, randomized, double-blind, parallel-groups, placebo-controlled, multicenter trials in outpatients with Major Depressive Disorder were performed. Aprepitant 160 mg and placebo were included in all trials. Aprepitant 80 mg and paroxetine 20 mg (active comparator) were included in three trials. Approximately 150 patients were enrolled per treatment group in each trial. The primary end point was the mean change-from-baseline of the first 17 items of the Hamilton Rating Scale for Depression (HAM-D(17)) score at 8 weeks. A positron emission tomography (PET) study was also performed in normal subjects to determine the relationship between neurokinin(1) receptor occupancy and aprepitant plasma concentrations in dosing regimens relevant to the trials. RESULTS: No statistically significant differences from placebo on the HAM-D(17) were observed at week 8 for either dose of aprepitant in any of the trials, whereas paroxetine 20 mg was significantly (p 2 cm. The median survival in patients with hysterectomy was 36 months and in those without hysterectomy 29 months, this difference was statistically not significant (P = 0.2). CONCLUSIONS: Our study indicates that in an unselected group of PPC patients 33% have any macroscopic uterine involvement. The therapeutic value of routine hysterectomy at the initial operation for PPC should be further investigated. PMID- 16249020 TI - Laparoscopy-assisted vaginal pelvic exenteration. AB - OBJECTIVE: The aim of this study was to evaluate the feasibility, morbidity and survival outcome of laparoscopy-assisted vaginal pelvic exenteration. METHODS: Since 2000, we have performed 5 cases of en-bloc pelvic exenteration combining a vaginal or perineal approach and laparoscopic approach. All patients had received previous pelvic irradiation. One patient underwent a total type II exenteration with ileal-loop diversion, an omental flap and a temporary colostomy. Two patients underwent a middle and posterior exenteration: one was a type III exenteration with perineal rectal resection and a gracilis myocutaneous flap; the second one was a type II exenteration with a colorectal anastomosis and a vaginal reconstruction using a gluteal thigh flap. Two patients underwent a type I anterior and middle exenteration with continent Miami pouch and vaginal reconstruction by omental cylinder. RESULTS: Mean time of the procedure was 6 h (range: 4.5-9). Peroperative bleeding was less than 500 cm3. Two patients presented minor complications: a perineal abscess after perineal rectal resection and an abdominal wound abscess. Mean length of hospital stay was 27 days. Three patients are free of disease. Two patients presented groin metastasis. One patient died of disease after 8 months. CONCLUSION: Laparoscopic or laparoscopy assisted vaginal pelvic exenteration followed by reconstruction is feasible with curative intent in selected patients. PMID- 16249021 TI - Surgically (laparotomy/laparoscopy) guided placement of high dose rate interstitial irradiation catheters (LG-HDRT): technique and outcome. AB - OBJECTIVE: To describe and evaluate the technique and the clinical outcome of a new modality for the treatment of women with persistent or recurrent pelvic malignancies utilizing surgically (laparotomy or laparoscopic) guided high dose rate (HDR) catheters to complete high dose rate interstitial irradiation therapy (LG-HDRT). METHODS: Between 6/2000 and 6/2004, 14 women with histologic evidence of postradiation persistent (3 patients) or recurrent (11 patients) pelvic disease underwent LG-HDRT. Five patients (36%) received treatment for a 2nd, 3rd or 4th recurrence. Preoperative clinical and radiologic evaluation to exclude evidence of extrapelvic disease was routine. Initial intraoperative evaluation included intraabdominal inspection and or biopsy to determine the extent of disease. A two "team" approach was used to place the 100 cm Teflon after loading HDR catheters. Each catheter had its open ends closed with bone wax prior to placement. Using a 14 gauge intravenous catheter as a guide, each HDR catheter was individually placed transvaginally. The tumor bed (treatment volume) was marked circumferentially with clips to facilitate treatment planning. Dosimetry was typically completed on the day of surgery and HDR therapy was started within the initial 24 postoperative hours. The catheters were removed transvaginally, without anesthesia following completion of therapy. RESULTS: Mean patient age was 63.1 years and weight was 138.2 lb. Squamous cell cancer of the vagina or cervix was the most common (64%) diagnosis. The mean time from initial diagnosis to LG HDRT was 67.9 months. The procedure was completed laparoscopically in 71% of patients, with 4 patients requiring laparotomy (3 conversions from laparoscopy). The mean duration of surgery was 94.9 min and the mean hospital stay was 4.8 days. Only 2 patients (14%) were discharged prior to the completion of therapy. The mean number of catheters placed was 6.1 and the mean dose delivered was 20 Gy over a mean of 5 fractions. There were no major intraoperative complications. Postradiation complications were limited to DVT (1), bladder bleeding (1), 6 months after response to a platinum-based regimen. The RECIST criteria were used for parameters of response. Tirapazamine was administered at a dose of 390 mg/m2 IV over 2 h followed 1 h later by cisplatin 60 mg/m2 IV every 3 weeks until disease progression or adverse effects prohibited further therapy. RESULTS.: Between June 2001 and February 2004, 65 patients were entered onto this study by 27 institutions; one patient was excluded due to ineligible tumor type. Twenty-six patients (41%) received six or more cycles of therapy; however, 16 (25%) received one course of therapy (mainly due to side effects or patient request). There were six (9%) complete responders and 28 (44%) partial responders for a total response rate of 53%. Only two patients (3%) developed increasing disease on this protocol, and response could not be assessed in 18 patients (28%). The median progression-free and overall survival for all patients is 10.9 and 26.4 months, respectively. The regimen did not cause major hematologic toxicity, however, it did cause frequent constitutional (23%) and gastrointestinal (mostly nausea/vomiting) (44%) grade 3 or 4 toxicity. CONCLUSIONS: The combination of tirapazamine and cisplatin has definite activity in the treatment of recurrent platinum-sensitive ovarian or primary peritoneal cancer. However, toxicity, primarily non-hematologic, was substantial. Reducing the toxicity of a tirapazamine-platinum combination should be pursued in future trials. PMID- 16249024 TI - Who is playing doctor? -- The gap between self-perceived versus professionally diagnosed chronic conditions among the underserved minority. AB - OBJECTIVE: To assess the gap between self-perceived and professionally diagnosed physical and mental conditions in underserved minority populations. STUDY SETTING: The study sample consisted of 287 African-American and Latino heads of households respondents, obtained from a geographically defined random sample of 418 households from three Urban Public Housing Communities in Los Angeles County, California. STUDY DESIGN: This study conducted a cross-sectional survey with face to-face interviews. RESULTS: Eighty-five percent (245) of this sample reported that they were suffering from at least one chronic condition. However, 43% of participants who suffer from chronic condition(s) claimed that a physician had never diagnosed at least one of their chronic illnesses. Depression (16%), hearing impairment (10%), arthritis (8%), dental problems (7%), and vision conditions (7%) are the most frequently reported undiagnosed chronic conditions among these populations. CONCLUSION: This study documents significant disparities in the detection and treatment of specific medical conditions among Hispanic and African-American population residing in public housing, which often leads to end stage complications due to lack of timely diagnoses and treatment. PMID- 16249025 TI - Unassisted recovery from stuttering: self-perceptions of current speech behavior, attitudes, and feelings. AB - The purpose of this study was to investigate the nature of recovery from stuttering based on the experiences of adults who recovered without treatment. Using a semi-structured, open-ended interview format, 15 speakers verified as persons who recovered without treatment were asked to describe their status as everyday speakers. Seven speakers reported that they no longer stuttered and eight reported that they still stuttered on occasion. Interview material was coded and analyzed by the investigators and checked by independent judges. Results suggested that complete recovery was possible for speakers who reported that they no longer stuttered; whereas, those who still stuttered occasionally appeared to no longer be handicapped by stuttering, but required some vigilance to maintain their relatively fluent speech. EDUCATIONAL OBJECTIVES: After completing this activity, the learner will be able to: (1) describe the relevance of self-report data for evaluating the nature of recovery from stuttering without treatment; (2) describe the differences in self-perception concerning the nature of recovery for those who no longer have any tendency to stutter compared to those who still have an occasional tendency to stutter; and (3) suggest the possible implications for understanding the nature of recovery from persistent stuttering based on investigations of late recovery without treatment. PMID- 16249026 TI - Metalloproteinases and human placental invasiveness. AB - Matrix metalloproteinases (MMPs) are the main mediators of extracellular matrix degradation. This confers an important role to these enzymes in the invasion of the trophoblast cells where their expression are spatiotemporally regulated. The regulation of MMPs activity is complex and is established at different levels. Their expressions depend on various cis-elements in their gene promoter, and are induced or repressed by various soluble factors. After expression and secretion, the proMMPs must be activated through an activation network. Then, the enzyme activity is regulated by inhibition or stabilization. In this review, we shall focus on the expression, the role in invasion and the regulation of MMPs in the human placenta. PMID- 16249027 TI - The neurologic complications in pediatric acute lymphoblastic leukemia patients excluding leukemic infiltration. AB - This study presents retrospective analyses of 20 acute lymphoblastic leukemia (ALL) patients who developed neurologic complications (except leukemic infiltration). These subjects represent 9.9% of 203 ALL patients aged 16 years or younger followed in our hospital between March 1991 and January 2003. Fourteen male and six female patients, whose ages ranged between 6 and 168 months, developed 24 episodes of neurologic complications after the diagnosis of ALL. The most common complication was meningitis, which developed in six (25%) episodes, and two thirds of the patients who had meningitis were evaluated to be iatrogenic. Cerebral infarct and venous thrombosis were detected in five (21%) of the episodes. In two (8%) episodes progressive cerebral dysfunction developed after radiotherapy. The remaining 11 (45%) episodes are due to varying types of complications. Interestingly, one patient had abundant histiocytes exhibiting hemophagocytosis in the cerebrospinal fluid (CSF) examination and this patient was subsequently diagnosed with fungal meningoencephalitis, by further investigation. Six (30%) patients died; epilepsia developed in five (25%) patients in the follow-up period and the remaining nine (45%) are healthy. By close follow-up and effective treatment of thrombosis and, especially, of infections including iatrogenic meningitis in developing countries, the morbidities and mortalities of these complications can be decreased. PMID- 16249028 TI - Different gamma/delta T clones sustain GVM and GVH effects in multiple myeloma patients after non-myeloablative transplantation. AB - TCR gamma/delta profiles were analyzed in 13 multiple myeloma patients after allogeneic non-myeloablative transplantation. Results show that both aGVHD and minimal residual disease (MRD) eradication did significantly affect TCR gamma/delta profile. During follow-up, six patients developed an aGVHD episode; in five of them, this event fitted with a modification of the TCR profile. Eleven patients achieved PCR-negativity during follow-up. In the 90% of them, the appearance of a new predominant TCR peak was concomitant to the disappearance of the IgH clone. These results suggest that different T gamma/delta populations would sustain GVM and GVH effects after non-myeloablative allogeneic transplant. PMID- 16249029 TI - MicroRNA modulation of megakaryoblast fate involves cholinergic signaling. AB - MicroRNAs (miRNAs) are abundant small regulatory RNAs with multiple roles in cell fate determination. The processes regulating cellular miRNA levels are still unclear and experimental oligonucleotide tools to readily mimic their effects are not yet available. Here, we report that thapsigargin-induced intracellular Ca(++) release suppressed pre-miR-181a levels in human promegakaryotic Meg-01 cells, induced differentiation-associated nuclear endoreduplication and caspase-3 activation and replaced the acetylcholinesterase 3' splice variant AChE-S with AChE-R. AChE, PKC and PKA inhibitors all attenuated the pre-miR-181a decline and the induced differentiation. AChmiON, a synthetic 23-mer 2'-oxymethylated oligonucleotide mimicking the miR-181a sequence, blocked the calcium-induced differentiation while elevating cellular pre-miR-181a levels and inducing DNA fragmentation and cell death. Moreover, when added to RW 264.7 macrophages, AChmiON at 100 nM induced nitric oxide production with efficiency close to that of bacterial endotoxin, demonstrating physiologically relevant activities also in blood-born monocytes/macrophages. The stress-induced modulation of hematopoietic miR-181a levels through AChE, PKC and PKA cascade(s) suggests using miRNA mimics for diverting the fate of hematopoietic tumor cells towards differentiation and/or apoptosis. PMID- 16249030 TI - Induction of atrazine tolerance in a natural soil assemblage of microalgae reared in the laboratory. AB - We investigated the occurrence of tolerance to atrazine (2-chloro-4-ethylamino-6 isopropylamino-s-triazine) in soil microalgae by means of the pollution-induced community tolerance methodology. To this end, a natural soil assemblage of microalgae, reared under laboratory conditions, was used as experimental model. Experimental cultures were exposed to 0.46, 0.93, 1.85, 2.70, and 5.40 mg atrazine/L medium for 40 days. After this chronic exposure, both untreated and atrazine-pretreated cultures were subjected to a short-term dose-effect study with atrazine using average growth rate as endpoint. Results showed that chronic exposure to atrazine induced significant changes at the community level, increasing its atrazine tolerance. In addition, atrazine-pretreated assemblages seemed to display, on average, a reduced growth rate compared with untreated assemblages in the absence of atrazine. Response to the presence of atrazine could thus be achieved with some fitness cost. This suggests that the soil microalgal productivity could be diminished after exposure to atrazine. PMID- 16249031 TI - Effects of copper and its ethylenediaminetetraacetate complex on the antioxidant defenses of the goldfish, Carassius auratus. AB - Copper bioavailability in the tissues of goldfish and antioxidant defenses in the liver of fish were investigated in vivo following 40 days of exposure to different species of copper solutions at different concentrations. Copper seemed to be more concentrated in the gill than in the liver after Cu(2+) and copper ethylenediaminetetraacetate (Cu-EDTA) exposure. The addition of EDTA markedly affected copper adsorption in the gill but had no significant effects in the liver. We also found that in the fish exposed to the concentration of Cu(2+) below the Chinese national fishery water quality standard of 0.01 mg/L, superoxide dismutase (SOD) and glutathione S-transferase (GST) activities in liver were stimulated and catalase (CAT) and Se-dependent glutathione peroxidase (Se-GPx) activities were inhibited after 40 days of exposure. Compared to copper exposure, the addition of EDTA did not affect SOD and GST significantly. However, CAT and GPx activity with Cu(2+) exposure were significantly different from those with Cu-EDTA exposure. PMID- 16249032 TI - Effects of methamidophos on acetylcholinesterase activity, behavior, and feeding rate of the white shrimp (Litopenaeus vannamei). AB - The toxicity of methamidophos on the shrimp Litopenaeus vannamei was evaluated using acetylcholinesterase (AChE) activity, behavior, and feeding rate as effect criteria. The biochemical characterization of the soluble cholunesterase (ChE) present in both muscle and eye tissues of L. vannamei was performed in a first phase of the study. In both tissues, almost full inhibition of enzyme activity by eserine sulfate was found, indicating that the measured activity is mainly from ChE and not from other esterases. The highest rate of substrate hydrolysis was found when acetylthiocholine was used as substrate. To evaluate the effects of methamidophos on L. vannamei AChE, behavior, and feeding rate, shrimps were exposed for 24h to several sublethal concentrations of methamidophos. Significant effects of the pesticide on behavior and AChE were found, with behavior being a more sensitive endpoint than AChE inhibition. Feeding rate was not a sensible endpoint under conditions tested. PMID- 16249033 TI - Growth factors, glucose and insulin kinetics after low dose growth hormone therapy in HIV-lipodystrophy. AB - OBJECTIVES: Low-dose growth hormone (GH) administration has been suggested as a treatment for HIV-lipodystrophy. METHODS: Postglucose GH-secretion, kinetics of insulin-like growth factors (IGFs), insulin, and glucose metabolism were examined in six male HIV-infected lipodystrophic patients (two normal-weight patients with normal glucose-tolerance (NGT), two normal-weight with impaired glucose-tolerance (IGT), and two obese patients with diabetes (DM)) during a 16 weeks open-labelled pilot-study of low-dose GH, 0.7 mg/day. RESULTS: DM, compared to NGT and IGT, displayed an impaired rebound of GH during a 5h oral glucose-tolerance test. Near lower normal limits in all patients before GH-therapy, total and free IGF-I increased between 87 and 152% during the GH-therapy (P<0.001), approaching upper normal limits in all patients with the highest incremental percentages shown in DM. A slight and temporary reduction in insulin sensitivity was caused by a reduction in non-oxidative glucose metabolism (n=5). GH-administration reduced hepatic extraction of insulin alleviating the demand for insulin secretion (n=5). No adverse effects of GH were detected. CONCLUSIONS: As judged from effects on circulating IGF-I, glucose metabolism, and insulin kinetics, 0.7 mg/day of GH may be expedient for treatment of HIV-infected males with lipodystrophy. Whether the patients' glucose metabolic status matters for the IGF-response to low-dose GH therapy awaits further investigation. PMID- 16249034 TI - A rare case of Brucella melitensis infection in an obstetrician during the delivery of a transplacentally infected infant. AB - We report a case of Brucella melitensis infection in an obstetrician who was infected during the delivery of an infant suffering from congenital brucellosis. The obstetrician was treated with doxycycline and rifampin and fully recovered. This is the first reported case of Brucella infection transmitted through infectious secretions at the delivery of a transplacentally infected newborn and emphasizes that, especially in endemic areas educational and technical measures are needed in order the obstetricians to avoid ingestion of secretions during clearance of the newborn's respiratory tract from saliva and amniotic fluid. PMID- 16249035 TI - Subjective life satisfaction and objective functional outcome in bipolar and unipolar mood disorders: a longitudinal analysis. AB - BACKGROUND: Quality of life (QOL) has gained increasing attention as an important yet underappreciated component of functional outcome in mood disorders. In particular, the relationship between subjective life satisfaction and objective measures of psychosocial adjustment has not been well-studied. The goal of the present study was to examine the longitudinal associations between subjective life satisfaction and objective functional outcome among individuals with bipolar and unipolar mood disorders. METHOD: One hundred fifty-seven mood disordered subjects were assessed at index hospitalization for bipolar mania (n=35), unipolar psychotic depression (n=27), or unipolar nonpsychotic depression (n=95). All were prospectively followed up three times, at approximately 2, 4.5 and 7-8 years. Global outcome, work performance, social adjustment, recurrent depressive episodes, and dimensions of life satisfaction were assessed by semi-structured interviews using standardized ratings. RESULTS: Subjective life satisfaction strongly paralleled global functioning, work performance and social adjustment at each follow-up for patients with unipolar nonpsychotic depression, but not bipolar disorder or unipolar psychotic depression. Depressive symptoms and objective functional impairment contributed to poor QOL in most domains, independent of illness chronicity, medication use, or affective disorder subtype. LIMITATIONS: Findings might have differed had a different QOL measure been used, although the present measure showed concurrent validity with a previously used instrument. Sample sizes for the bipolar and psychotic depression groups were sufficient to detect moderate, but not small, correlations between objective functioning and subjective QOL. CONCLUSIONS: Recurrent depression remains a substantial contributor to poor life satisfaction across affective disorder subtypes. Subjective QOL in bipolar and unipolar psychotic depression patients may not accurately reflect objective functional outcome status, potentially due to diminished insight, demoralization, or altered life expectations over time. PMID- 16249036 TI - Regulated expression of peripheral node addressin-positive high endothelial venules controls seeding of B lymphocytes into developing neonatal rabbit appendix. AB - Young rabbit appendix is a homologue of chicken bursa of Fabricius; both are crucial sites for preimmune B-cell repertoire diversification. Here, we report that appendix regulates precursor lymphocyte recruitment for further development by modulating the sites of extravasation. The total area of peripheral node addressin-positive (PNAd(+)) high endothelial venules (HEVs) increased from 1 day to 1 week after birth, remained constant up to 2 weeks and declined to a low and persistent amount by 3 weeks. In normal 1-week and manipulated 5-week appendix where growth of follicles was retarded, PNAd(+) HEVs were present in the basolateral sides of B-cell follicles whereas, in normal 5-wk-appendix these were restricted to T-cell areas. The PNAd was expressed on the lumenal surface of HEVs. The proportions of CD62L(+) B cells in appendix declined from approximately 40% at 3 days to 2-3% at 4 weeks. In lymphocyte transfer experiments, CD62L(+) B cells were preferentially recruited compared with CD62L(-) B cells, anti-PNAd antibody blocked migration of B cells by approximately 50%, and 100 times more B cells were recruited in 1-week compared to 6-week appendix. Thus, a unique spatiotemporal expression pattern of PNAd(+) HEVs is associated with development of B-cell follicles. This regulates migration of blood-borne B-lymphocytes into developing appendix by interacting with CD62L. PMID- 16249037 TI - Methamphetamine stimulates interferon inducible genes in HIV infected brain. AB - OBJECTIVE: To study the synergism of HIV and methamphetamine. DESIGN AND METHODS: We undertook a microarray study using RNA from the frontal cortex of 15 individuals with HIV infection to initially identify genes that are differentially regulated by HIV encephalitis (HIVE). From the analysis of the microarray data, we identified candidate genes to be validated by quantitative real time PCR (qRT-PCR) and to assess if these genes were differentially modulated in individuals with HIVE and documented methamphetamine use. RESULTS: Analysis of microarray data revealed that genes involved in several categories were dysregulated in HIVE. We then chose 15 candidate genes for validation by qRT PCR and analyzed the tissue concentration of these genes across three groups: those with HIV infection and no brain pathology, those with HIVE, and those with both HIVE and a history of methamphetamine use. We noted that there was upregulation of interferon inducible genes in the HIVE with methamphetamine using group, which together as a gene group was highly statistically significant (p=0.0064). CONCLUSION: These findings indicate that dysregulation of interferon inducible genes may underlie the pathogenic mechanism resulting in greater neurodegenerative and neurocognitive burden that occurs in methamphetamine using HIV infected individuals. PMID- 16249038 TI - Differential protein expression profiles in anterior gills of Eriocheir sinensis during acclimation to cadmium. AB - Using a proteomic approach, we characterized different protein expression profiles in anterior gills of the Chinese mitten crab, Eriocheir sinensis, after cadmium (Cd) exposure. Two experimental conditions were tested: (i) an acute exposure (i.e. 500 microg Cd l(-1) for 3 days) for which physiological, biochemical and ultrastructural damage have been observed previously; (ii) a chronic exposure (i.e. 50 microg Cd l(-1) for 30 days) resulting in physiological acclimation, i.e. increased resistance to a subsequent acute exposure. Two dimensional gel electrophoresis (2-DE) revealed six protein spots differentially expressed after acute, and 31 after chronic Cd exposure. From these spots, 15 protein species were identified using MS/MS micro-sequencing and MS BLAST database searches. Alpha tubulin, glutathione S-transferase and crustacean calcium-binding protein 23 were down-regulated after an acute exposure, whereas another glutathione S-transferase isoform was up-regulated. Furthermore, analyses revealed the over-expression of protein disulfide isomerase, thioredoxin peroxidase, glutathione S-transferase, a proteasome subunit and cathepsin D after chronic exposure. Under the same condition, ATP synthase beta, alpha tubulin, arginine kinase, glyceraldehyde-3-phosphate dehydrogenase and malate dehydrogenase were down-regulated. These results demonstrate that acute and chronic exposure to waterborne Cd induced different responses at the protein expression level. Protein identification supports the idea that Cd mainly exerts its toxicity through oxidative stress induction and sulfhydryl-group binding. As a result, analyses showed the up-regulation of several antioxidant enzymes and chaperonins during acclimation process. The gill proteolytic capacity seems also to be increased. On the other hand, the clearly decreased abundance of several enzymes involved in energy transfer suggests that chronic metal exposure induced an important metabolic reshuffling. PMID- 16249039 TI - Altered cardiovascular responses to chronic angiotensin II infusion in aged rats. AB - In this work we determined by telemetry the cardiovascular effects produced by Ang II infusion on blood pressure (BP) and heart rate (HR) in aged rats. Male Wistar aged (48-52 weeks) and young (12 weeks) rats were used. Ang II (6 microg/h, young, n=6; aged, n=6) or vehicle (0.9% NaCl 1 microl/h, young, n=4; aged, n=5) were infused subcutaneously for 7 days, using osmotic mini-pump. The basal diurnal and nocturnal BP values were higher in aged rats (day: 98+/-0.3 mm Hg, night: 104+/-0.4 mm Hg) than in the young rats (day: 92+/-0.2 mm Hg, night: 99+/-0.2 mm Hg). In contrast, the basal diurnal and nocturnal HR values were significantly smaller in the aged rats. Ang II infusion produced a greater increase in the diurnal BP in the aged rats (Delta MAP=37+/-1.8 mm Hg) compared to the young ones (Delta MAP=30+/-3.5 mm Hg). In contrast, the nocturnal MAP increase was similar in both groups (young rats; Delta MAP=22+/-3.0 mm Hg, aged rats; Delta MAP=24+/-2.6 mm Hg). During Ang II infusion HR decreased transiently in the young rats. An opposite trend was observed in the aged rats. Ang II infusion also inverted the BP circadian rhythm, in both groups. No changes in HR circadian rhythm were observed. These differences suggest that the aging process alters in a different way Ang II-sensitive neural pathways involved in the control of autonomic activity. PMID- 16249040 TI - Does gender modify the detrimental coronary effects of metabolic syndrome? AB - The association between metabolic syndrome and coronary artery disease, either clinical or subclinical, has been well established. There is also a consensus regarding the higher prevalence of metabolic syndrome in females compared with males with coronary artery disease. However, the impact of gender on the coronary artery disease severity has not been investigated in patients with metabolic syndrome. The available data have suggested no significant association between gender and the extent and severity of coronary artery disease. PMID- 16249041 TI - Kounis syndrome (allergic angina and allergic myocardial infarction): a natural paradigm? AB - Inflammatory mediators including histamine, neutral proteases, arachidonic acid products, platelet activating factor and a variety of cytokines and chemokines are increased in blood or urine in both allergic episodes and acute coronary syndromes. The release of mediators during allergic insults has been incriminated to induce coronary artery spasm and/or atheromatous plaque erosion or rupture. A common pathway between allergic and non-allergic coronary syndromes seems to exist. Today, there is evidence that mast cells not only enter the culprit region before plaque erosion or rupture but they release their contents before an actual coronary episode. Kounis syndrome is the concurrence of acute coronary syndromes with conditions associated with mast cell activation including allergic or hypersensitivity and anaphylactic or anaphylactoid insults. It is caused by inflammatory mediators released through mast cell activation. Kounis syndrome, as consequence, of the above pathophysiologic association is regarded as nature's own experiment and magnificent natural paradigm showing novel way in an effort to prevent acute coronary syndromes. Drugs and natural molecules which stabilize mast cell membrane and monoclonal antibodies that protect mast cell surface could emerge as novel therapeutic modalities capable to prevent acute coronary and cerebrovascular events. PMID- 16249042 TI - Cardiac tamponade in Hashimoto's disease. AB - Myxedematous pericardial effusions, occurring in about one-third of patients with hypothyroidism, usually do not cause symptoms and regress slowly with hormonal treatment. We report a cardiac tamponade inaugural of a Hashimoto's disease. PMID- 16249043 TI - Detection of volatile metabolites produced by bacterial growth in blood culture media by selected ion flow tube mass spectrometry (SIFT-MS). AB - To achieve faster bacteremia diagnosis, selected ion flow tube mass spectrometry (SIFT-MS) measured metabolic gases in the headspaces of BacT/ALERT blood culture bottles. Pseudomonas aeruginosa, Streptococcus pneumoniae, Escherichia coli, Staphylococcus aureus and Neisseria meningitidis growth and trace gas patterns were detected from 10 colony forming units after 6 h. PMID- 16249044 TI - Characterization of ORF2 and its encoded protein of the Helicoverpa armigera nucleopolyhedrovirus. AB - The open reading frame 2 (ha2) of the Helicoverpa armigera single nucleocapsid nucleopolyhedrovirus (HaSNPV), a conserved gene in most baculoviruses from lepidopteran insects such as p78/83 of the Autographa californica MNPV, was characterized. It is 1,242 bp long and potentially encodes a 45.9 kDa. Ha2 is conserved among baculoviruses from lepidopteran insects. Ha2 transcripts were detected from 16 to 96 h post infection (hpi) of HzAM1 cells. Rabbit polyclonal antiserum against a GST-HA2 fusion protein reacted with three protein of 50, 46 and 35 kDa at 24-72 hpi of HzAM1 cells. Anti OpMNPV ORF2 (homologue of HA2) antibody reacted only with the 46 and 35 kDa proteins in HaSNPV-infected cells. These results demonstrate that Ha2 is modified at the mRNA or protein levels. Western blot analysis showed that only the 50 kDa product of HA2 is a structural component of proteins of both the budded virus (BV) and occlusion-derived virus (ODV) phenotypes. HA2-EGFP fusion protein showed that HA2 is localized primarily in the nucleus of HzAM1 infected cells. The HA2 was found to co-localize with actin by labelling of actin with Rhordamine-Phalloidin. In summary, the data indicated that HA2 is a structural protein and interacts with host cell actin. PMID- 16249045 TI - Measurement of contaminant removal from skin using a portable fluorescence scanning system. AB - The residence time of particulate contamination on the human body is a factor that has an important impact on the accuracy of exposure assessment in the aftermath of an accidental release of radionuclides to the atmosphere. Measurements of particle clearance from human skin were made using an illumination system to excite fluorescence in labelled silica particles and a CCD camera and image processing system to detect this fluorescence. The illumination system consists of high-intensity light emitting diodes (LEDS) of suitable wavelengths arranged on a portable stand. The physically small size of the LEDs allows them to be positioned close to the fluorescing surface, thus maximising the fluorescent signal that can be obtained. The limit of detection was found to be 50 microg of tracer particle per cm2. Experiments were carried out to determine the clearance rates of 10 microm and 3 microm particles from the skin. Results show that, in the absence of any mechanical rubbing of the skin, the clearance of particles from the skin followed an approximately exponential decay with a half-time of 1.5-7.8 h. Skin hairiness and degree of human movement were found, in addition to particle size, to have an important influence on particle fall-off rate. PMID- 16249046 TI - Trace metals in sediments of two estuarine lagoons from Puerto Rico. AB - Concentrations of As, Cd, Cu, Fe, Hg, Pb and Zn were evaluated in surface sediments of two estuaries from Puerto Rico, known as San Jose Lagoon (SJL) and Joyuda Lagoon. Significantly higher concentrations in microg/g dw of Cd (1.8 vs. 0.1), Cu (105 vs. 22), Hg (1.9 vs. 0.17), Pb (219 vs. 8), and Zn (531 vs. 52) were found in sediment samples from SJL when compared to Joyuda Lagoon. Average concentrations of Hg, Pb, and Zn in some sediment samples from SJL were above the effect range median (ERM) that predict toxic effects to aquatic organisms. Enrichments factors using Fe as a normalizer, and correlation matrices showed that metal pollution in SJL was the product of anthropogenic sources, while the metal content in Joyuda Lagoon was of natural origins. Sediment metal concentrations found in SJL were comparable to aquatic systems classified as contaminated from other regions of the world. PMID- 16249047 TI - Relation between PAH and black carbon contents in size fractions of Norwegian harbor sediments. AB - Distributions of total organic carbon (TOC), black carbon (BC), and polycyclic aromatic hydrocarbons (PAH) were investigated in different particle size fractions for four Norwegian harbor sediments. The total PAH (16-EPA) concentrations ranged from 2 to 113 mg/kg dry weight with the greatest fraction of PAH mass in the sand fraction for three of the four sediments. TOC contents ranged from 0.84% to 14.2% and BC contents from 0.085% to 1.7%. This corresponds to organic carbon (OC = TOC - BC) contents in the range of 0.81-14% and BC:TOC ratios of 1.3-18.1%. PAH isomer ratios suggested that the PAH in all four sediments were of pyrogenic origin. Furthermore, stronger correlations between PAH versus BC (r2 = 0.85) than versus OC (r2 = 0.15) were found. For all size fractions and bulk sediments, the PAH-to-BC ratios for the total PAHs were on average 6+/-3 mg PAH/g BC. These results suggest that PAH distributions were dominated by the presence of BC, rather than OC. As sorption to BC is much stronger than sorption to OC, this may result in significantly lower dissolved concentrations of PAH than expected on the basis of organic carbon partitioning alone. PMID- 16249048 TI - An analysis of the proposed REACH regulation. AB - In October 2003, the European Commission produced its first draft of its new chemicals legislation, known as REACH. At its core, REACH will require producers and users of chemical substances to register any use in a volume-triggered system. Mandatory submission of chemical assessment reports containing information on the hazards, exposures and risks associated with the uses of the chemical substances will be reviewed by government appointed expert committees. The marketing of substances considered to be of very high concern will require authorisation. This paper analyses issues related to the architecture of the proposed REACH regulation in light of its origins, drivers, its impacts on businesses and possible unintended consequences on other industries outside the chemical industry. Since the design of REACH reflects a range of different sources, goals and ideas, a number of its provisions are ambiguous in their current form. This creates uncertainty as to the implementation of the regulation. Regulatory certainty is, however, an essential pre-condition for the effective functioning of a modern market economy. As currently drafted, REACH could pose a challenge to the operation of the market economy in the EU. PMID- 16249049 TI - Social class assignment and mortality in Sweden. AB - The earlier practice of assigning all members of a family to the same social class as that of the household head, typically the father, has in recent years been replaced by either basing individual class position on one's own occupation or of one of the family members, not necessarily the father. These various practices have been extensively scrutinised for more than 20 years. The validity of the approaches has chiefly been tested by checking how well they account for the variation in some criteria, mostly class identification, political attitudes and voting behaviour. Here it is shown, using census data from Sweden, that mortality-rate differences between social classes covering the period 1991-1997 are greater for both men and women when both spouses are assigned to the same social class on the basis of the dominance approach, where the labour market position of either spouse may determine the social class of the family. It is suggested that the common observation that class differences are smaller among women than among men may, at least to some extent, be the result of establishing a woman's class position on the basis of her own occupation rather than the labour market position of her spouse. PMID- 16249050 TI - The functional role of the subthalamic nucleus in cognitive and limbic circuits. AB - Once it was believed that the subthalamic nucleus (STN) was no more than a relay station serving as a "gate" for ascending basal ganglia-thalamocortical circuits. Nowadays, the STN is considered to be one of the main regulators of motor function related to the basal ganglia. The role of the STN in the regulation of associative and limbic functions related to the basal ganglia has generally received little attention. In the present review, the functional role of the STN in the control of cortico-basal ganglia-thalamocortical associative and limbic circuits is discussed. In the past 20 years the concepts about the functional role of the STN have changed dramatically: from being an inhibitory nucleus to a potent excitatory nucleus, and from being involved in hyperkinesias to hypokinesias. However, it has been demonstrated only recently, mainly by reports on the behavioral (side-) effects of STN deep brain stimulation (DBS), which is a popular surgical technique in the treatment of patients suffering from advanced Parkinson Disease (PD), that the STN is clinically involved in associative and limbic functions. These findings were confirmed by results from animal studies. Experimental studies applying STN DBS or STN lesions to investigate the neuronal mechanisms involved in these procedures found profound effects on cognitive and motivational parameters. The anatomical, electrophysiological and behavioral data presented in this review point towards a potent regulatory function of the STN in the processing of associative and limbic information towards cortical and subcortical regions. In conclusion, it can be stated that the STN has anatomically a central position within the basal ganglia thalamocortical associative and limbic circuits and is functionally a potent regulator of these pathways. PMID- 16249051 TI - Diagnosis of Trichomonas vaginalis infection: The sensitivities and specificities of microscopy, culture and PCR assay. AB - OBJECTIVES: The aim of this study was to compare wet mount-, Giemsa stain-, acridine orange fluorescent stain-, cultivation- and polymerase chain reaction (PCR)-based approaches to establish which method or combination of methods was most effective in the laboratory diagnosis of trichomoniasis. STUDY DESIGN: Out of 200 investigated patients with various gynecological complaints, Trichomonas vaginalis infection was detected in 27 (13.5%) by any of methods investigated. Among women with trichomonads, a typical clinical finding was presented in only nine. For analysis of sensitivity and specificity of the methods used, the receiver operating characteristic (ROC) curve concept with culture as a gold standard was applied. RESULTS: Infection was diagnosed by wet mount in 14 (7.0%) women, by Giemsa stain in 11 (5.5%) and by acridine orange stain in 16 (8.0%) women. In 21 (10.5%) women, it was diagnosed by culture in Diamond's medium, and in 22 (11.0%) by PCR. For the initial diagnosis of trichomoniasis, wet preparation is the test that is widely available in most STD clinics, but its sensitivity is poor (66.67%). Giemsa stain shows a low sensitivity of 52.38%. Acridine orange shows reasonable sensitivity and specificity of 71.43% and 99.44%, respectively. The sensitivity and specificity of PCR (80.95% and 97.21%) did not exceed that of culture. CONCLUSION: With regard to the fact that trichomoniasis can have an atypical or even asymptomatic course, in order to accurately diagnose this disease, microbiological investigation is necessary. Comparison of different methods showed that at least two techniques, such as culture and acridine orange staining, have the potential for better diagnosis of T. vaginalis infection. PCR detection of infection has been demonstrated to be highly specific and sensitive, but its availability and cost effectiveness are in question. PCR could provide an alternative for laboratory diagnosis of trichomoniasis by culture. PMID- 16249052 TI - Duration of HPV-associated risk for high-grade cervical intraepithelial neoplasia. AB - OBJECTIVE: To evaluate the duration of high-risk HPV-associated cancer risk. STUDY DESIGN: Patients who had had a routine diagnostic Hybrid Capture Tube Test (HCT) due to squamous cell abnormalities of the uterine cervix were followed-up until the endpoint of histologically diagnosed cervical intraepithelial neoplasia (CIN). RESULTS: Six hundred and thirty-eight women were followed during a cumulative follow-up of 16,423 patient months. The adjusted relative risk associated with the positive HR-HCT test for high-grade CIN/52.0 (20.9-19.2)/ proved to be higher than that of the cytological atypia/5.44 (2.52-11.77)/. At the end of the 30 months of follow-up the crude and adjusted risks for CIN2+ were 214.3 (28.4-1615.7) and 196.7 (25.4-1525.2), respectively in the HPV 16/18 group, and after 30 months, the crude and adjusted RR decreased to 57.6 (10.4-318.9) and 29.2 (5.02-170.0). In the groups of other high-risk types and possibly high-risk types the general tendency was the same. However, new CIN2+ cases were not detected after the 30th month of follow-up in these later groups. CONCLUSIONS: HPV16/18 associated relative risk is nearly 200 times higher than that of the HPV negative population and an outstanding risk persists with duration of about 30 months. The risk is manifested in progression to high-grade CIN lesions mainly within a 2 years interval after the first detection of HPV 16/18 infection. PMID- 16249053 TI - Elevation of telomerase activity positively correlates to poor prognosis of patients with non-small cell lung cancer. AB - Increased telomerase activity has been found in various types of human malignancies, including lung cancer. However, the correlation between the level of telomerase activity and the clinical characteristics of lung cancer patients remains unclear. The levels of telomerase activity in lung cancer specimens and adjacent non-neoplastic tissues obtained from 68 patients who underwent surgery were measured by using a non-radioactive quantitative method. Clinical and pathologic parameters were evaluated with respect to the level of telomerase activity. Prominent telomerase activity was detected in 58 (85.3%) lung cancer tissues and 21 (30.9%) adjacent non-neoplastic tissues. There was a trend of increase in relative telomerase activity in regard to the advanced pathological stage, and lymph node metastasis. Using Cox regression analysis, we found that every 100 unit of increase in relative telomerase activity was associated with an increase in the hazard ratio of death by 13% after controlling for other variables such as age, gender, and stage (Hazard ratio=1.13; 95% CI: 1.03-1.23, P=0.006). For patients with stage I disease, an increase of every 100unit of relative telomerase activity was associated with an even higher increase of 33% in the hazard ratio of death (Hazard ratio=1.33; 95% CI: 1.07-1.65, P=0.011) and a 16% increase in the hazard of disease recurrence (Hazard ratio=1.16, 95% CI: 1.01-1.33, P=0.032). The level of telomerase activity is positively correlated with the risk of recurrence and mortality of lung cancer. The level of telomerase activity would predict the prognosis of lung cancer patients. PMID- 16249054 TI - Tonic neuronal activation during simple and complex finger movements analyzed by DC-magnetoencephalography. AB - Functional neuroimaging techniques map neuronal activation indirectly via local concomitant cortical vascular/metabolic changes. In a complementary approach, DC magnetoencephalography measures neuronal activation dynamics directly, notably in a time range of the slow vascular/metabolic response. Here, using this technique neuronal activation dynamics and patterns for simple and complex finger movements are characterized intraindividually: in 6/6 right-handed subjects contralateral prolonged (30 s each) complex self-paced sequential finger movements revealed stronger field amplitudes over the pericentral sensorimotor cortex than simple movements. A consistent lateralization for contralateral versus ipsilateral finger movements was not found (4/6). A subsequent sensory paradigm focused on somatosensory afferences during the motor tasks and the reliability of the measuring technique. In all six subjects stable sustained neuronal activation during electrical median nerve stimulation was recorded. These neuronal quasi tonic activation characteristics provide a new non-invasive neurophysiological measure to interpret signals mapped by functional neuroimaging techniques. PMID- 16249055 TI - Role of peptide primary sequence in polyphenol-protein recognition: an example with neurotensin. AB - Polyphenols are known for their impact on health and one of their major properties is the formation of complexes with proteins. To investigate the involvement of polyphenol-protein complexes in health, the interactions between bioactive polyphenols and neurotensin were examined by structural NMR and molecular modeling. Neurotensin is a linear bioactive tridecapeptide and polyphenols seem to affect the NT metabolism. We studied the polyphenols resveratrol and its glucoside the piceid in order to observe the possible role of glucose group and the penta-O-galloyl-D-glucopyranose (PGG). NMR data and molecular modeling showed that interaction occurred with the three polyphenols involving hydrophobic stacking and hydrogen bonds. Moreover, the peptide primary sequence plays a role in the specificity of complex formation. PMID- 16249056 TI - CtIP, a candidate tumor susceptibility gene is a team player with luminaries. AB - CtIP is a nuclear protein conserved among vertebrates that was discovered as a cofactor of the transcriptional corepressor CtBP. CtIP also interacts with the tumor suppressors such as BRCA1 and the pRb family members through binding sites that are frequently mutated in human cancers. CtIP is a target for BRCA1 dependent phosphorylation by the ATM kinase induced by DNA double strand breakage. CtIP plays a role in DNA-damage-induced cell cycle checkpoint control at the G2/M transition. Homozygous inactivation of the Ctip gene causes very early embryonic lethality during mouse development. The Ctip(-/-) embryo cells are arrested in G1 and do not enter S phase. Depletion of Ctip in established mouse embryo fibroblasts arrests cells in G1 and results in an accumulation of hypophosphorylated Rb and the Cdk inhibitor p21, suggesting that CtIP is also a critical regulator of G1/S transition of the cell cycle. The Ctip gene contains a mononucleotide (A9) repeat and one of the alleles is mutated at a high frequency in colon cancers with microsatellite instability. The Ctip(+/-) mice develop multiple types of tumors suggesting that haploid insufficiency of Ctip leads to tumorigenesis. Among the various tumor types observed in Ctip(+/-) heterozygous mice, large lymphomas are prevalent. Recent studies raise the possibility that Ctip may itself be a tumor susceptibility gene and suggest that it might be important for the activities of tumor suppressors BRCA1, pRb family proteins and Ikaros family members. PMID- 16249057 TI - Moving into the future: treatment of bone metastases and beyond. AB - In various common cancers, the skeleton is a preferred site of metastasis. It is also threatened by bone loss resulting from anti-cancer therapy. Like bone metastases in advanced cancer, cancer treatment induced bone loss (CTIBL) substantially increases fracture risk and dramatically decreases quality of life and patient autonomy. Both chemotherapy and endocrine deprivation therapy (EDT) can significantly decrease bone mineral density (BMD). This is aggravated by the often long duration of EDT, particularly in the adjuvant setting. Cumulative bone loss can put patients at risk of osteopenia and osteoporosis. With their known efficacy in preventing skeletal complications in metastatic disease, bisphosphonates (BP) lend themselves to exploring their extended use, notably in preventing CTIBL. Clinical trials have shown BPs to effectively prevent and treat CTIBL, for which they are recommended by ASCO guidelines. Whether BPs also have the potential to prevent metastatic dissemination to bone remains to be determined. Zoledronic acid, a third-generation BP with a favourable efficacy/safety record, was shown by ongoing large clinical trials to not only prevent or reduce CTIBL in early stage cancer, but to actually increase BMD. The current evidence of the potential of zoledronic acid in addressing CTIBL and preventing bone metastases is reviewed. PMID- 16249058 TI - Effects of paroxetine and venlafaxine on immune parameters in patients with obsessive compulsive disorder. AB - BACKGROUND: Obsessive-compulsive disorder (OCD) has been associated with an altered activity of the immune system. This study was carried out to investigate whether treatment with paroxetine and venlafaxine modifies the immune function in OCD and whether this modification is related to treatment outcome. METHODS: Ex vivo production of TNF-alpha, IL-4, IL-6, IL-10, and IFN-gamma in whole blood cultures, and NK-cell activity and peripheral blood NK-cell-, monocytes-, T-cell , and B-cell- percentages were measured in 42 outpatients with OCD who participated in a 12-week, double-blind SRI trial. RESULTS: Paroxetine and venlafaxine treatment did not affect immune parameters and clinical response was not directly related to altered activity of the immune system. Responders, defined as having a >or=25% decrease on the Y-BOCS, differed from non-responders with regard to the change of TNF-alpha values. CONCLUSIONS: The present results suggest that paroxetine and venlafaxine do not affect immune parameters and that clinical response is not related to altered activity of the immune system in OCD. Nonetheless, our data yielded interesting preliminary results that warrant further discussion and investigation. PMID- 16249059 TI - Role of inter and intramolecular interactions in protein-DNA recognition. AB - Protein-DNA recognition plays an essential role in the regulation of gene expression. Regulatory proteins are known to recognize specific DNA sequences directly through atomic contacts between protein and DNA, and/or indirectly through the conformational properties of the DNA. In this work, we have analyzed the specificity of intermolecular interactions by statistical analysis of base amino acid interactions within protein-DNA complexes as well as the computer simulations of base-amino acid interactions. The specificity of the intramolecular interactions was studied by statistical analysis of the sequence dependent DNA conformational parameters and the elastic properties of DNA. Systematic comparison of these specificities in a large number of protein-DNA complexes revealed that both intermolecular and intramolecular interactions contribute to the specificity of protein-DNA recognition, and their relative contributions vary depending upon the protein-DNA complex. We demonstrated that combination of the intermolecular and intramolecular energies leads to enhanced specificity and the combined energy could explain experimental data on binding affinity changes caused by base mutations. These results provided new insight into the relationship between specificity and structure in the process of protein DNA recognition, which would lead to prediction of specific protein-DNA binding sites. PMID- 16249060 TI - Use of mucosal immunization with porcine zona pellucida (PZP) in mice and rabbits. AB - Rabbits (Oryctolagus cuniculus) and two strains of mice (Mus musculus, one inbred and one outbred) were immunized against porcine zona pellucida (PZP) antigen. Alginate microspheres or cholera toxin B were used alone or in combination when mucosal immunization routes were used. Serum antibody responses and fertility were assessed. Neither rabbit or mouse groups immunized by mucosal routes generated significant antibody responses to PZP as compared to parenteral immunization (ANOVA, P > 0.05). The study shows that porcine zona pellucida is not an effective mucosal antigen in small mammals. PMID- 16249061 TI - Effects of the Kielmeyera coriacea extract on energy metabolism in the rat liver. AB - Kielmeyera coriacea Mart is a medicinal plant of the Clusiacea (Guttiferae) family used by the native population of Brazil in the treatment of several tropical diseases such as malaria, schistosomiasis, leishmaniasis, and fungal or bacterial infections. Kielmeyera coriacea is also effective as an antidepressant drug. Extracts of the plant are rich in xanthones. Compounds of this class have been reported to inhibit mitochondrial energy metabolism. For this reason the action of the Kielmeyera coriacea extract on hepatic energy metabolism was investigated in the present work, using isolated rat liver mitochondria and the perfused rat liver. In perfused livers the extract (20-80 microg/ml) caused stimulation of oxygen consumption, inhibition of gluconeogenesis and stimulation of glycogenolysis and glycolysis. In isolated mitochondria the Kielmeyera coriacea extract (5-20 microg/ml) stimulated state IV respiration, reduced the ADP/O ratio and decreased the respiratory coefficient. The activities of succinate-oxidase, NADH-oxidase, NADH dehydrogenase and succinate dehydrogenase were inhibited. The ATPase of intact mitochondria was stimulated and the ATPase of uncoupled mitochondria was inhibited. The results of this investigation suggest that the Kielmeyera coriacea extract impairs the hepatic energy metabolism by acting as mitochondrial uncoupler and inhibitor of enzymatic activities linked to the respiratory chain. The impairment of mitochondrial energy metabolism could lead to adverse metabolic effects by the use of the crude extract, but it could equally be the basis of its antiprotozoan and antifungal effects. PMID- 16249062 TI - Clinical suspicion as a primary guidance to use commercial amplification tests for rapid diagnosis of pulmonary tuberculosis. AB - The Abbott LCx (Abbott Park, IL) Mycobacterium tuberculosis complex is a commercial amplification assay discontinued from the European market in 2002. A prospective clinical study was carried out to evaluate the clinical utility of the above test as applied by specialists for the rapid diagnosis of active pulmonary tuberculosis (PTB). According to the physician's clinical judgment, patients were classified into 3 groups (low, intermediate, and high) aiming to estimate the probability of active disease. The gold standard for final diagnosis was based on microbiologic and clinical information including data from a 6-month follow-up period. Sensitivities and specificities of rapid microbiologic tests were compared with those based on an integrated approach including clinical evaluation plus the above tests. The incidence of PTB in 214 patients was 13.1%. The basis for initial treatment of PTB was smear-positive results in 46%, positive LCx results in 29%, and clinical suspicion in 18%. For the remaining 7%, therapy was started upon receipt of culture results. The sensitivity, specificity, and positive and negative predictive values of the LCx assay were 68%, 99%, 95%, and 95%, respectively. In comparison, they were 93%, 99%, 96%, and 99%, respectively, for the combination of clinical evaluation plus the LCx test. It is concluded that in patients with high-to-moderate pretest probabilities, the combination of clinical judgment and amplification results strongly enhances a rapid and correct diagnosis of PTB. PMID- 16249063 TI - Genetic characterization of Vibrio cholerae isolates from Argentina by V. cholerae repeated sequences-polymerase chain reaction. AB - We have developed a novel typing method based on Vibrio cholerae repeat sequences (VCR) using primers directed out of the VCR sequences. To evaluate the VCR polymerase chain reaction (PCR) as a typing system, 2 categories, efficacy and efficiency, were analyzed in 69 strains of human and environmental V. cholerae O1 toxigenic and nontoxigenic, and non-O1 strains isolated since 1992-2000 from Argentina. The discriminatory power (0.91), stability (0.95), reproducibility (1), typeability (1), rapidity, accessibility, as well ease of use, indicated that the VCR-PCR method provides an alternative useful tool for molecular epidemiology of V. cholerae. The VCR-PCR of V. cholerae isolates showed 29 patterns, of which pattern 1 represented 68% of the V. cholerae O1 isolates, supporting the hypothesis that a clone with epidemic behavior was responsible for the epidemic in Latin America. These results showed a good correlation and a better epidemiologic analysis when the results were compared in parallel with repetitive extragenic palindromic sequences-PCR. In conclusion, VCR-PCR showed excellent performance as a typing method for cholera surveillance programs. PMID- 16249064 TI - Mycobacterium abscessus infection after use of tumor necrosis factor alpha inhibitor therapy: case report and review of infectious complications associated with tumor necrosis factor alpha inhibitor use. AB - Tumor necrosis factor alpha (TNF-alpha) inhibitors, such as infliximab and etanercept, are now frequently used in the treatment of inflammatory diseases including rheumatoid arthritis (RA) and Crohn's disease. As an apparent result of their immune modulating activity, there has been an observed association between the use of these agents and the development of a wide range of infections, most notably Mycobacterium tuberculosis. We describe a case of infection with Mycobacterium abscessus in a 67-year-old woman receiving infliximab as a component of her therapy for RA. This case, along with extensive reports in the medical literature, illustrate how treatment with inhibitors of TNF-alpha has the potential to result in a wide range of infectious complications, including rapid growing Mycobacterium. PMID- 16249065 TI - Rapid diagnosis and genotyping of Leishmania isolates from cutaneous and visceral leishmaniasis by microcapillary cultivation and polymerase chain reaction restriction fragment length polymorphism of miniexon region. AB - We have performed a combination of microcapillary cultivation method and restriction fragment length polymorphism (RFLP) analysis of amplified products by 1 single PCR of miniexon region of Leishmania for molecular diagnosis and genotyping of different Leishmania species isolated from cutaneous leishmaniasis (CL) and visceral leishmaniasis. We have analyzed 10 microcapillary cultivated isolates from cutaneous cases and 5 microcapillary cultivated isolates from visceral cases (totally 15) by polymerase chain reaction-RFLP (PCR-RFLP). Of 10 isolates, 3 (30%) were genotyped as Leishmania infantum and 7 (70%) of 10 isolates were genotyped as Leishmania tropica from the microcapillary cultivated isolates of cutaneous cases. On the other hand, all 5 isolates (100%) were genotyped as L. infantum from microcapillary cultivated visceral cases. Our most interesting finding is the presence of 3 L. infantum isolates in CL cases without kala-azar history. Therefore, we suggest that further investigations must be done about this subject. On the other hand, we suggest the combination of microcapillary culture method and PCR-RFLP of miniexon region of leishmaniae can be used in routine laboratory experimentation because of their simple, cheap, and rapid benefits (within a week), whereas other different approaches offer a multitude of valid taxonomic characters for species identification. PMID- 16249066 TI - Targeted, haplotype-resolved resequencing of long segments of the human genome. AB - Currently, challenges exist to acquire long-range (hundreds of kilobase pairs) phase-discriminated sequence across substantial numbers of individuals. We have developed a straightforward method for isolating and characterizing specific genomic regions in a haplospecific manner. Real-time PCR is carried out to STS content map and genotype pools of fosmid clones arrayed in 384-well microtiter plates. Single-nucleotide polymorphisms, microsatellite markers, and insertion deletion polymorphisms are used to differentiate the target region into haplotype specific tiling paths. DNA of clones from these tiling paths is retrieved from the library and either sequenced by standard shotgun methods or amplified in vitro and sequenced by a primer-based, directed method. This approach provides convenient access to complete, haplotype-resolved resequencing data from multiple individuals across tens to hundreds of thousands of basepairs. We illustrate its implementation with a detailed example of more than 400 kbp from the human CFTR region, across 15 individuals, and summarize our experience applying it to many other human loci. PMID- 16249067 TI - Ameliorating effect of folic acid on chromium(VI)-induced changes in reproductive performance and seminal plasma biochemistry in male rabbits. AB - Chromium hexavalent (Cr(VI)) is a biologically active oxidized state of chromium. It is involved in the redox cycle, with the production of reactive oxygen species. Free radical scavenging properties and possible antioxidant activity of folic acid (FA) have been reported; therefore, the present study examined possible protective effects of FA on the reproductive toxicity of potassium dichromate (K2Cr2O7) in male New Zealand white rabbits. We monitored reproductive performance, lipid peroxidation, enzyme activities and biochemical parameters in seminal plasma. Six rabbits per treatment group (and a control group) were exposed: 8.3 microg/kg FA; 5 mg/kg potassium dichromate (contains 3.6 mg chromium(VI)) and 5 mg/kg potassium dichromate+8.3 microg/kg FA. Results showed that semen quality deteriorated following potassium dichromate exposure. Testosterone levels, body weight (BW), relative weights of testes (RTW) and epididymis (REW) all decreased. Levels of thiobarbituric acid-reactive substances increased, whereas the activities of glutathione S-transferase, transaminases and phosphatases decreased in the seminal plasma. FA alone significantly increased BW, RTW, REW, semen characteristics and seminal plasma enzymes, and decreased the levels of free radicals. Furthermore, FA can be effective in the protection of chromium-induced reproductive toxicity. PMID- 16249068 TI - An assessment of the potential developmental and reproductive toxicity of di isoheptyl phthalate in rodents. AB - Di-isoheptyl phthalate (DIHP) is a branched, phthalate ester with seven carbon alkyl side chains. Since structurally similar phthalates have been shown to produce developmental and/or reproductive effects in rodents, the potential for DIHP to produce developmental and reproductive toxicity was assessed. In a developmental toxicity study, female rats were given DIHP by oral gavage on gestational days 6-20. There were significant reductions in uterine weight, increased resorptions and reduced fetal weight in the high dose (750 mg/kg) group. Fetal examination revealed malformations and variations of both the skeletal system and the viscera including ectopic testes. The intermediate dose, 300 mg/(kg/day), was a no effect level in this study. In a two-generation reproductive toxicity study, DIHP was given in the diet at 1000, 4500 and 8000 ppm. In the 8000 ppm group of the first (F1) generation, anogenital distance was reduced, time to balanopreputial separation was increased, there was a significant increase in thoracic nipples and testicular abnormalities, and weights of testes and accessory reproductive organs were significantly reduced. Testicular sperm counts and daily sperm production were significantly reduced. Fertility was also significantly reduced in the 8000 ppm group. In the second (F2) generation offspring, anogenital distance was significantly reduced and there was evidence of reduced weight gain during lactation in both the 4500 and 8000 ppm groups. The overall no effect level (NOEL) in the reproductive toxicity study was in the range of 64-168 mg/(kg/day) (gestation-lactation periods). By comparison, estimated average human exposures in the general population are <1 microg/(kg/day). PMID- 16249069 TI - In vivo molecular imaging of adhesion molecules in experimental autoimmune encephalomyelitis (EAE). AB - The infiltration of autoreactive T cells into the central nervous system (CNS) requires a complex molecular interplay between immune cells and the blood-brain barrier (BBB), especially involving adhesion molecules like intercellular adhesion molecule (ICAM)-1. Here we study the molecular expression at the BBB during adoptively transferred (AT) myelin basic protein (MBP)-experimental autoimmune encephalomyelitis (EAE) in vivo by sensitive particle acoustic quantification (SPAQ)-enhanced ultrasound after intravenous application of specific gas-filled MP (MP) targeted against ICAM-1 (ICAM-MP) as contrast agent. Our results reveal a clear periventricular and cerebellar upregulation of ICAM-1 expression at the disease maximum of AT-EAE. Moreover, SPAQ-enhanced ultrasound enables the sensitive quantification of ICAM-1 expression in vivo. This allows to monitor therapeutic changes as shown by suppression of ICAM-1 expression after pretreatment of rats with corticosteroids (P < 0.008). All imaging results were confirmed by parallel immunohistochemistry. In vivo magnetic resonance imaging and albumin staining of rat brains after sonification did not reveal a disturbance of the BBB, thereby proving the safety of the method. Subsequent application of specific MP did not influence follow-up measurements, a prerequisite for sequential measurements in longitudinal studies. Based on these data, quantitative molecular imaging of adhesion molecules by SPAQ-enhanced ultrasound proves to be a safe and reliable technology to monitor changes at the BBB in vivo. PMID- 16249070 TI - Comparative study of GAD65-specific CD4+ T cells in healthy and type 1 diabetic subjects. AB - Glutamic acid decarboxylase 65 (GAD65) is a putative autoantigen associated with the pathogenesis of type 1 diabetes (T1D). The prevalence of autoreactive CD4+ T cells towards the immunodominant GAD65(555-567) epitope in DR4 healthy and T1D subjects was investigated with class II tetramers. A slightly higher percentage of diabetic subjects had GAD65(555-567) tetramer-positive T cells upon GAD65(555 567) peptide stimulation on the total CD4+ T-cell populations compared to healthy subjects. In contrast, three quarters of subjects in both groups had tetramer positive T cells resulting from stimulation of the CD4+CD25+ regulatory T-cell depleted CD4+ T cells. The frequencies and TCR Vbeta gene usages of GAD65(555 567) T cells were also similar in both groups. Experiments demonstrated that GAD65(555-567)-reactive T cells in healthy and diabetic subjects had different CD45RA phenotypes. For the healthy group, GAD65(555-567)-reactive T cells were generally found in the CD45RA+ naive T-cell pool while GAD65(555-567)-reactive T cells from T1D subjects were present in both CD45RA+ naive and CD45RA- memory T cell pools. These findings suggested that there is no difference in thymic selection of DR4 restricted GAD-reactive T cells amongst healthy and T1D individuals but GAD65(555-567)-reactive T cells have been preferentially activated in diabetic patients. PMID- 16249071 TI - Identification of tear lipocalin as a novel autoantigen target in Sjogren's syndrome. AB - Sjogren's syndrome (SS) is an autoimmune disease characterized by lymphocytic infiltration and tissue damage mainly confined to the salivary and lacrimal glands, resulting in dryness of mouth and eyes. Since different epithelial cells of exocrine and non-exocrine tissues are primarily affected, an autoimmune reaction against antigens commonly expressed in epithelial cells is believed to play a pathogenic role. To identify novel autoantigen targets associated with the systemic involvement in SS, we screened a random peptide library with pooled IgG immunoglobulins derived from patients with primary SS. Among the identified peptides, one was recognized by the majority of patients' sera, but not by sera of normal donors and of patients with other autoimmune diseases. The peptide showed homology with an Epstein-Barr Virus (EBV) derived protein and with tear lipocalin, a protein highly expressed in tears and saliva, and with alpha-fodrin, a cytoskeleton protein considered an important autoantigen target in SS. Anti peptide antibodies affinity purified from patients' sera recognize the viral protein, tear lipocalin and alpha-fodrin. Our findings suggest that EBV infection may be linked to the pathogenesis of SS and that tear lipocalin can be considered a novel and yet unidentified autoantigen in SS. PMID- 16249072 TI - Clinical guidelines for the management of neutropenic patients with unexplained fever in Japan: validation by the Japan Febrile Neutropenia Study Group. AB - The Japan Febrile Neutropenia Study Group (JFNSG) Trial was a multicenter, open, randomized study designed to validate the first Japanese guidelines for the management of neutropenic cancer patients with unexplained fever issued in 1998. The trial compared cefepime monotherapy with cefepime plus amikacin combination therapy in febrile neutropenic patients with hematological disorders. The JFNSG found that monotherapy with cefepime was, in general, as effective as combination therapy. In terms of subset analyses, defervescence appeared to occur more frequently in leukemic patients and in those with profound neutropenia treated with the dual combination. The conclusion of the trial was that the 1998 guidelines were applicable to the Japanese febrile neutropenic patient population. The JFNSG met again in 2003 to revise these guidelines. An important addition to the guidelines was a distinction between low- and high-risk patients. Low-risk febrile neutropenic patients can receive oral ciprofloxacin or levofloxacin, with or without amoxicillin/clavulanic acid, on an outpatient basis, or intravenous (i.v.) monotherapy with cefepime, ceftazidime or a carbapenem. High-risk patients can receive i.v. cefepime, ceftazidime or a carbapenem, or an i.v. dual combination with cefepime, ceftazidime or a carbapenem plus an aminoglycoside. Those patients with a documented infection with methicillin-resistant Staphylococcus aureus should also receive a glycopeptide. It remains to be determined whether existing assessment scoring systems apply to Japanese patients; whether a broad-spectrum cephalosporin plus an aminoglycoside combination is required as the initial management of patients with acute leukemia and/or profound neutropenia; which antibacterial drugs should be used when first- and second-line agents fail; what are the appropriate oral agents and dosing regimens for low-risk patients; whether serology or the polymerase chain reaction should be the preferred marker for initiating preemptive antifungal therapy; and whether the azoles or the candins should be the preferred antifungal agents. PMID- 16249073 TI - Placebo-controlled trial of agomelatine in the treatment of major depressive disorder. AB - The efficacy and safety of flexible dosing with the antidepressant agomelatine (25-50 mg/day) was evaluated in a 6-week, double-blind, randomized, placebo controlled study involving 212 patients who met Diagnostic and Statistical Manual of Mental Disorders-Fourth Edition (DSM-IV) criteria for major depressive disorder-current major depressive episode. Patients receiving agomelatine (25 mg and 50 mg/day) had a significantly lower mean Hamilton Rating Scale for Depression (HAM-D) score at endpoint compared with those who received placebo (14.1 +/- 7.7 vs. 16.5 +/- 7.4, p = 0.026). Agomelatine significantly improved the response rate (49.1%; p = 0.03), time to first response (p = 0.032), and Clinical Global Impression-Severity of Illness score (p = 0.017), compared with placebo. These results were confirmed in a subgroup of patients with greater symptom severity. Agomelatine 50 mg also appeared to be effective and well tolerated in patients who failed to show improvement after 2 weeks on a dose of 25 mg/day. These results support the prescription of agomelatine 25 mg as the usual therapeutic dose, and suggest that increasing the dose to 50 mg may be beneficial for some patients without reducing tolerability. PMID- 16249074 TI - CYCLCROP indirect 13C mapping for long-time and chemical-shift selective diffusion measurements in complex hydrocarbon systems. AB - Cyclic cross-polarization from a proton magnetization to 13C and from there back to proton coherences permits the indirect, 13C chemical shift selective detection of hydrocarbon compounds in the proton NMR channel. This excitation technique can be combined with elements of one-, two- or three-dimensional magnetic resonance imaging permitting the measurement of time-resolved spatial distributions of hydrocarbon components. Beginning this sort of CYCLCROP mapping experiment with a non-equilibrium distribution of the constituents in the system allows one to study the time evolution of the concentrations of all components that can be identified by characteristic 13C resonance lines. As applications, studies of ingress, mixing, gel formation, transport and metabolism in living plants, long time inter- and self-diffusion in complex hydrocarbon systems are suggested. As a test experiment, the diffusion of methanol in swollen polymethylmethacrylate was examined. PMID- 16249076 TI - Indigenous healers' explanations of low back pain and its relief. AB - Significant number of Finns consults annually indigenous healers for their low back pain (LBP). Bone setting is the most vital traditional Finnish mobilization treatment of LBP. In the present study we have clarified healers' believes on the pathophysiology of back pain and observed them at work to clarify the effectiveness of treatment by medical check-up and by interviewing the patients who also compared the benefits of this method with their previous other treatments such as drugs, exercise and physiotherapy. STUDY DESIGN: Two of the oldest bone setters participated in the study. In the patient follow-up they treated 35 consecutive chronic LBP subjects. A medical check-up of the patients was done before and after their treatments. After 7 months mean follow-up time patients replied by using questionnaires and evaluated the effectiveness of the method. RESULTS: The indigenous bone setters told that they followed the hundreds of years old tradition. The healers explained that the back pain was due to difference in the leg lengths and this was reflected in the pelvic and spinal asymmetry as well as increased muscle tension. After the sessions the subjects reported relaxation and clear immediate pain alleviation. The patients reported in the follow-up questionnaires benefits from their LBP treatment in 28 cases out of 29 who replied after the treatment. According to physician's evaluation 27 patients (from 35 or 77%) got excellent or good result in their chronic LBP. There was no change in one case, and none got worse. Bone setting received in patients' reports better evaluation than drug treatment and physical therapy. PMID- 16249075 TI - Mediofrontal negativities in the absence of responding. AB - The feedback-related negativity (FRN) is an event-related brain potential component that is elicited by feedback stimuli indicating unfavorable outcomes. Until recently, the FRN has been studied primarily using experimental paradigms in which outcomes appeared to be contingent upon the participants' behavior. The present study further addressed the question whether an FRN can be elicited by outcomes that are not contingent on any preceding choice or action. Participants took part in a simple slot-machine task in which they experienced monetary gains and losses in the absence of responses. In addition, they performed a time estimation task often used to study the FRN and a flanker task known to elicit the error-related negativity. Outcomes in the slot-machine task elicited an FRN like mediofrontal negativity whose amplitude correlated with the amplitude of the FRN associated with negative feedback in the time estimation task. However, the mediofrontal negativity was observed both for (unfavorable) outcomes that averted a gain and for (favorable) outcomes that averted a loss of money. The results are discussed in the framework of current conceptions of the FRN and related electrophysiological components. PMID- 16249077 TI - A formal theory for spatial representation and reasoning in biomedical ontologies. AB - OBJECTIVE: The objective of this paper is to demonstrate how a formal spatial theory can be used as an important tool for disambiguating the spatial information embodied in biomedical ontologies and for enhancing their automatic reasoning capabilities. METHOD AND MATERIALS: This paper presents a formal theory of parthood and location relations among individuals, called Basic Inclusion Theory (BIT). Since biomedical ontologies are comprised of assertions about classes of individuals (rather than assertions about individuals), we define parthood and location relations among classes in the extended theory Basic Inclusion Theory for Classes (BIT+Cl). We then demonstrate the usefulness of this formal theory for making the logical structure of spatial information more precise in two ontologies concerned with human anatomy: the Foundational Model of Anatomy (FMA) and GALEN. RESULTS: We find that in both the FMA and GALEN, class level spatial relations with different logical properties are not always explicitly distinguished. As a result, the spatial information included in these biomedical ontologies is often ambiguous and the possibilities for implementing consistent automatic reasoning within or across ontologies are limited. CONCLUSION: Precise formal characterizations of all spatial relations assumed by a biomedical ontology are necessary to ensure that the information embodied in the ontology can be fully and coherently utilized in a computational environment. This paper can be seen as an important beginning step toward achieving this goal, but much more work along these lines is required. PMID- 16249078 TI - Exploiting naturally occurring DNA variation and molecular profiling data to dissect disease and drug response traits. AB - Identifying the key drivers of common human diseases and associated signaling pathways remains one of the primary objectives in the biomedical and life sciences. In this respect, common inbred strains of mice have played a crucial role, and recent advances in the development of genomics and bioinformatics tools have significantly enhanced their utility for this purpose. These advances have enabled a more holistic, network-oriented view of biological systems that facilitates elucidation of the underlying causes of disease and the best ways to target them. Success in reconstructing gene networks underlying disease traits (or other complex traits like drug response) and identifying the key drivers of these traits now largely rests on integrative approaches that combine data from multiple different sources. Such integrative genomics approaches that take into account genotypic, molecular profiling and clinical data in segregating mouse populations have recently been developed. Key to this integration has been the development and application of sophisticated algorithms to mine the diversity of data. PMID- 16249079 TI - Allosteric regulation of chaperonins. AB - Chaperonins are molecular machines that facilitate protein folding by undergoing energy (ATP)-dependent movements that are coordinated in time and space by complex allosteric regulation. Recently, progress has been made in describing the various functional (allosteric) states of these machines, the pathways by which they interconvert, and the coupling between allosteric transitions and protein folding reactions. However, various mechanistic issues remain to be resolved. PMID- 16249080 TI - Pregnancy, progesterone and progestins in relation to breast cancer risk. AB - In the last two decades the prevailing opinion, supported by the "estrogen augmented by progesterone" hypothesis, has been that progesterone contributes to the development of breast cancer (BC). Support for this opinion was provided by the finding that some synthetic progestins, when added to estrogen in hormone replacement therapy (HRT) for menopausal complaints, increase the BC risk more than estrogen alone. However, recent findings suggest that both the production of progesterone during pregnancy and the progesterone endogenously produced or exogenously administered outside pregnancy, does not increase BC risk, and could even be protective. The increased BC risk found with the addition of synthetic progestins to estrogen in HRT seems in all likelihood due to the fact that these progestins (medroxyprogesterone acetate and 19-nortestosterone-derivatives) are endowed with some non-progesterone-like effects which can potentiate the proliferative action of estrogens. The use of progestational agents in pregnancy, for example to prevent preterm birth, does not cause concern in relation to BC risk. PMID- 16249081 TI - Macrocyclic peptidomimetic inhibitors of beta-secretase (BACE): first X-ray structure of a macrocyclic peptidomimetic-BACE complex. AB - The synthesis of novel macrocyclic peptidomimetic inhibitors of the enzyme BACE1 is described. These macrocycles are derived from a hydroxyethylene core structure. Compound 7 was co-crystallized with BACE1 and the X-ray structure of the complex elucidated at 1.6 Angstrom resolution. This molecule inhibits the production of the Abeta peptide in HEK293 cells overexpressing APP751sw. PMID- 16249082 TI - A novel class of sodium/calcium exchanger inhibitors: design, synthesis, and structure-activity relationships of 4-phenyl-3-(piperidin-4-yl)-3,4-dihydro-2(1H) quinazolinone derivatives. AB - Design, synthesis, and structure-activity relationships of 3,4-dihydro-2(1H) quinazolinone derivatives as sodium/calcium (Na+/Ca2+) exchanger inhibitors are described. In these studies, optimization of the substituents at the 3-position of this series of compounds was carried out and dramatic effects of the substituent on the activities were observed. Based on these SAR studies, a highly potent inhibitor of the Na+/Ca2+ exchanger, which showed single-digit-nanomolar activity, was discovered. PMID- 16249083 TI - Macrolactonization to 10-deoxymethynolide catalyzed by the recombinant thioesterase of the picromycin/methymycin polyketide synthase. AB - The recombinant thioesterase (TE) domain of the picromycin/methymycin synthase (PICS) catalyzes the macrolactonization of 3, the N-acetylcysteamine thioester of seco-10-deoxymethynolide to generate 10-deoxymethynolide (1) with high efficiency. By contrast, 4, the 7-dihydro derivative of seco-thioester 3, undergoes exclusive hydrolysis by PICS TE to seco-acid 5. The recombinant TE domain of 6-deoxyerythronolide B synthase (DEBS TE) shows the same reaction specificity as PICS TE, but with significantly lower activity. PMID- 16249084 TI - Non-covalent modification of the heme-pocket of apomyoglobin by a 1,10 phenanthroline derivative. AB - To expand the repertoire of artificial enzymes that are constructed by replacing the natural prosthetic group of hemoproteins with non-natural cofactors, we examined incorporation of a non-porphyrinic ligand (1) into the heme-pocket of apomyoglobin in a non-covalent fashion. Ligand 1 is a highly conjugated 1,10 phenanthroline derivative, which shares some structural features with protoporphyrin IX; for example, molecular size and arrangement of hydrophobic and anionic parts. Addition of apomyoglobin to a solution of 1 induces clear changes in the absorption spectrum of 1, suggesting one-to-one incorporation of 1 into the heme cavity of apomyoglobin with an affinity of 6.3 x 10(6)M(-1). We found that the hydrolytic activity of apomyoglobin toward p-nitrophenyl hexanoate was greatly suppressed because of the incorporation of 1 into the heme-pocket. PMID- 16249085 TI - Pyrazoloheteroaryls: novel p38alpha MAP kinase inhibiting scaffolds with oral activity. AB - A test library with three novel p38alpha inhibitory scaffolds and a narrow set of substituents was prepared. Appropriate combination of substituent and scaffold generated potent p38alpha inhibitors, for example, pyrazolo[3,4-b]pyridine 9, pyrazolo[3,4-d]pyrimidine 18a and pyrazolo[3,4-b]pyrazine 23b with potent in vivo activity upon oral administration in animal models of rheumatoid arthritis. PMID- 16249086 TI - Asymmetries in the spatial distributions of enhancing lesions and black holes in relapsing-remitting MS. AB - Magnetic resonance imaging (MRI) is the most important paraclinical test in the diagnosis of multiple sclerosis (MS) and for delineating its natural history. We investigate MRIs from a longitudinal study of 24 relapsing-remitting MS patients who had monthly MRI examinations for one year, and were not receiving active MS therapy during this period. We hypothesized that lesions occur randomly throughout the brain, and that patients are homogeneous with regard to spatial patterns of lesion presentation. We recorded the numbers and locations of enhancing lesions and hypointense lesions (black holes) in all scans, and found asymmetrical patterns of lesions about the mid-transaxial, mid-coronal, and mid sagittal planes. Furthermore, in distinct subsets of patients, enhancing lesions and black holes tend to occur in the same locations. Clustering in lesion locations may be of functional significance, with consequent therapeutic implications. PMID- 16249087 TI - High-throughput screening of ecdysone agonists using a reporter gene assay followed by 3-D QSAR analysis of the molting hormonal activity. AB - In this study, 172 diacylhydrazine analogs were examined for their ability to activate an ecdysone (molting hormone)-dependent reporter gene in a silkworm (Bombyx mori) cell-based high-throughput screening assay. The measured EC(50) values (concentration required to cause an effect in 50% of the cells) were used to construct a 3-D QSAR model that describes the ecdysone agonist activities of the diacylhydrazine analogs. Of these compounds, 14 exhibited no activity and were excluded from the 3-D QSAR analysis. The resulting equation described approximately 74% of the activity for 158 compounds. The final equation consisted of 42% electrostatic and 58% steric effects (r(2) = 0.74 and q(2) = 0.45). Comparative molecular field analysis (CoMFA) was used to visualize the steric and electrostatic potential fields that were favorable and unfavorable for biological activity. Of particular interest was the observation that the hydrophobic parameter (logP) was not necessary for describing the observed activities, although previous studies have cited the importance of hydrophobic parameters in both classical and 3-D QSAR analyses of these compounds. Modeling studies of the B. mori ecdysone receptor supported the observed physicochemical parameters required for activity reported by the CoMFA models. Comparison of the present analysis with those performed using other lepidopteran assay systems evidenced a high degree of correlation (r(2) = 0.81 for a Sf-9 cell-based assay and r(2) = 0.89 for a Chilo suppressalis integument-based assay), indicating that it is valid to compare the results generated with the B. mori cell-based system to those generated with previous lepidopteran assays. This novel assay system is amendable to a high-throughput screening format and should greatly increase our ability to discover novel agonists of molting hormone (ecdysone) activity. PMID- 16249088 TI - Evaluation of aldose reductase inhibition and docking studies of some secondary metabolites, isolated from Origanum vulgare L. ssp. hirtum. AB - Five polar constituents of Origanum vulgare L. ssp. hirtum were investigated for their ability to inhibit aldose reductase (ALR2), the first enzyme of the polyol pathway implicated in the secondary complications of diabetes. The most active compound was found to be lithospermic acid B. Caffeic acid was inactive as it showed no inhibitory activity against the enzyme. The order of the inhibitory activity of the remaining compounds was: rosmarinic acid >12-hydroxyjasmonic acid 12-O-beta-glucopyranoside > p-menth-3-ene-1,2-diol 1-O-beta-glucopyranoside. Docking studies have been undertaken to gain insight into the binding mode of the investigated compounds at the active site of ALR2. The predicted hydrogen bonding and hydrophobic interactions may explain the observed inhibitory activity. PMID- 16249089 TI - Synthesis and evaluation of azaindole-alpha-alkyloxyphenylpropionic acid analogues as PPARalpha/gamma agonists. AB - A series of azaindole-alpha-alkyloxyphenylpropionic acid analogues was synthesized and evaluated for PPAR agonist activities. Structure-activity relationship was developed for PPARalpha/gamma dual agonism. One of the synthesized compound 7a was identified as a potent, selective PPARalpha/gamma dual agonist. PMID- 16249090 TI - Solid phase synthesis and antiprotozoal evaluation of di- and trisubstituted 5' carboxamidoadenosine analogues. AB - The rapid increase of resistance to drugs commonly used in the treatment of tropical diseases such as malaria and African sleeping sickness calls for the prompt development of new safe and efficacious drugs. The pathogenic protozoan parasites lack the capability of synthesising purines de novo and they take up preformed purines from their host through various transmembrane transporters. Adenosine derivatives constitute a class of potential therapeutics due to their selective internalisation by these transporters. Automated solid-phase synthesis can speed up the process of lead finding and we pursued the solid-phase synthesis of di- and trisubstituted 5'-carboxamidoadenosine derivatives by using a safety catch approach. While efforts with Kenner's sulfonamide linker remained fruitless, successful application of the hydrazide safety-catch linker allowed the construction of two representative combinatorial libraries. Their antiprotozoal evaluation identified two compounds with promising activity: N(6) benzyl-5'-N-phenylcarboxamidoadenosine with an IC(50) value of 0.91 microM against Trypanosoma brucei rhodesiense and N(6)-diphenylethyl-5' phenylcarboxamidoadenosine with an IC(50) value of 1.8 microM against chloroquine resistant Plasmodium falciparum. PMID- 16249091 TI - Structural basis for DNA-cleaving activity of resveratrol in the presence of Cu(II). AB - Resveratrol (1, 3,5,4'-trihydroxy-trans-stilbene), a polyphenol found in grapes and other food products, is known as an antioxidant and cancer chemopreventive agent. However, 1 was shown to induce genotoxicity through a high frequency of micronucleus and sister chromatid exchange in vitro and DNA-cleaving activity in the presence of Cu(II). The present study was designed to explore the structure activity relationship of 1 in DNA strand scission and to characterize the substrate specificity for Cu(II) and DNA binding. When pBR322DNA was incubated with 1 or its analogues differing in the number and positions of hydroxyl groups in the presence of Cu(II), the ability of 4-hydroxystilbene analogues to induce DNA strand scission is much stronger than that of 3-hydroxy analogues. The high binding affinity with both Cu(II) and DNA was also observed by 4-hydroxystilbene analogues. The reduction of Cu(II) which is essential for activation of molecular oxygen proceeded by addition of 1 to the solution of the Cu(II)-DNA complex, while such reduction was not observed with the addition of isoresveratrol, in which the 4-hydroxy group of 1 is changed to the 3-position. The results show that the 4-hydroxystilbene structure of 1 is a major determinant of generation of reactive oxygen species that was responsible for DNA strand scission. PMID- 16249092 TI - Advances in the synthesis and recent therapeutic applications of 1,2,4 thiadiazole heterocycles. AB - Chemical properties of 1,2,4-thiadiazole have been reviewed in the last few years. However, the usefulness of 1,2,4-thiadiazole as a privileged system in medicinal chemistry has prompted the advances on the therapeutic potential of this system. This review provides a brief summary of the medicinal chemistry of 1,2,4-thiadiazole system and highlights some examples of 1,2,4-thiadiazole containing drug substances in the current literature. A survey of representative literature procedures for the preparation of 1,2,4-thiadiazole is presented in sections by generalized synthetic methods. PMID- 16249093 TI - Substituted phenanthrenes with basic amino side chains: a new series of anti breast cancer agents. AB - In the course of our search for new anti-breast cancer agents, substituted phenanthrenes with basic amino side chains were synthesized and some of them showed remarkable antiproliferative activity against ER +ve MCF-7 cell line with IC(50) in the range of 3.53-22.25 microM. One of the compounds 15 ca showed anti breast cancer activity in 7,12-dimethylbenz[a]anthracene (DMBA) induced hormone dependent mammary tumor in rat and the activity was comparable to that shown by tamoxifen. PMID- 16249094 TI - Effect of polycyclic cage amines on the transmembrane potential of neuronal cells. AB - A series of pentacycloundecylamine derivatives were synthesized and their influence on the transmembrane potential of human SH-SY5Y neuroblastoma cells was evaluated using laser scanning confocal microscopy in combination with the potentiometric dye tetramethylrhodamine methyl ester. Results indicate that these derivatives influence the profile of KCl-induced membrane depolarization and cause an overall reduction in cell membrane depolarization. PMID- 16249095 TI - Structure-based design of isoquinoline-5-sulfonamide inhibitors of protein kinase B. AB - Structure-based drug design of novel isoquinoline-5-sulfonamide inhibitors of PKB as potential antitumour agents was investigated. Constrained pyrrolidine analogues that mimicked the bound conformation of linear prototypes were identified and investigated by co-crystal structure determinations with the related protein PKA. Detailed variation in the binding modes between inhibitors with similar overall conformations was observed. Potent PKB inhibitors from this series inhibited GSK3beta phosphorylation in cellular assays, consistent with inhibition of PKB kinase activity in cells. PMID- 16249096 TI - Group-II metabotropic glutamate receptors negatively modulate NMDA transmission at striatal cholinergic terminals: role of P/Q-type high voltage activated Ca++ channels and endogenous dopamine. AB - Striatal cholinergic nerve terminals express functional group-II metabotropic (mGlu) and NMDA glutamate receptors. To investigate whether these receptors interact to regulate ACh release, LY354740 (a group-II mGlu receptor agonist) and NMDA were co-applied in striatal synaptosomes and slices. LY354740 prevented the NMDA-evoked [3H]-choline release from synaptosomes and ACh release from slices. In synaptosomes, this modulation was prevented by omega-agatoxin IVA, suggesting that it was mediated by P/Q-type high voltage activated Ca++ channels. In slices, LY341495 (a group-II mGlu receptor antagonist) enhanced the NMDA-induced ACh release, suggesting that group-II mGlu receptor activation by endogenous glutamate inhibits NMDA transmission. Co-immunoprecipitation studies excluded direct group-II mGlu-NMDA receptor interactions. Finally, group-II mGlu negative modulation of NMDA transmission was abolished in dopamine-depleted synaptosomes and slices, suggesting that it relied on endogenous dopamine. We conclude that group-II mGlu receptors attenuate NMDA inputs at striatal cholinergic terminals via Ca++ channel modulation and dopamine-sensitive pathways. PMID- 16249097 TI - Production and purification of self-assembling peptides in Ralstonia eutropha. AB - Self-assembling peptides have emerged as an attractive scaffold material for tissue engineering, yet the expense associated with solid phase chemical synthesis has limited their broad use. In addition, the fidelity of chemical synthesis constrains the length of polypeptides that can be produced homogeneously by this method. Template-derived biosynthesis by recombinant DNA technology may overcome both of these problems. However, recovery of polypeptides from recombinant protein expression systems typically involves multi-step purification schemes. In this study, we report an integrated approach to recombinantly produce and purify self-assembling peptides from the recently developed expression host Ralstonia eutropha. The purification is based on the specific affinity of carbohydrate binding modules (CBMs) to cellulose. In a first step, we identified CBMs that express well in R. eutropha by assembling a fusion library of green fluorescent protein (GFP) and CBMs and determining the fluorescence of cell-free extracts. Three GFP::CBM fusions were found to express at levels similar to GFP alone, of which two CBMs were able to mediate cellulose binding of the GFP::CBM fusion. These two CBMs were then fused to multiple repeats of the self-assembling peptide RAD16-I::E (N-RADARADARADARADAE-C). The fusion protein CBM::E::(RAD16-I::E)4 was expressed in R. eutropha and purified using the CBM's affinity for cellulose. Subsequent proteolytic cleavage with endoproteinase GluC liberated RAD16-I::E peptide monomers with similar properties to the chemically synthesized counterpart RAD16-I. PMID- 16249098 TI - Neural correlates of regulating negative emotions related to moral violations. AB - Previous neuroimaging studies have identified several brain regions associated with regulating emotional responses. Different kinds of emotional stimuli, however, may recruit different regulatory processes and, in turn, recruit different regions. We compared emotion regulation for two types of negative emotional stimuli: those involving moral violations (moral stimuli), and those not involving moral violations (non-moral stimuli). In addition, we investigated whether activation in medial prefrontal cortex (MPFC), a region implicated previously in specifically moral processing, may instead reflect greater social and emotional content. Ten female subjects were scanned using fMRI while they passively viewed or were instructed to decrease emotional reactions to moral and non-moral pictures closely matched on social and emotional content. Passive viewing of both picture types elicited similar activations in areas related to the processing of social and emotional content, including MPFC and amygdala. During regulation, different patterns of activation in these regions were observed for moral vs. non-moral pictures. These results suggest that the neural correlates of regulating emotional reactions are modulated by the emotional content of stimuli, such as moral violations. In addition, the current findings suggest that some brain regions previously implicated in moral processing reflect the processing of greater social and emotional content in moral stimuli. PMID- 16249099 TI - Resolving fiber crossing using advanced fast marching tractography based on diffusion tensor imaging. AB - Magnetic resonance diffusion tensor tractography is a powerful tool for the non invasive depiction of the white matter architecture in the human brain. However, due to limitations in the underlying tensor model, the technique is often unable to reconstruct correct trajectories in heterogeneous fiber arrangements, such as axonal crossings. A novel tractography method based on fast marching (FM) is proposed which is capable of resolving fiber crossings and also permits trajectories to branch. It detects heterogeneous fiber arrangements by incorporating information from the entire diffusion tensor. The FM speed function is adapted to the local tensor characteristics, allowing in particular to maintain the front evolution direction in crossing situations. In addition, the FM's discretization error is reduced by increasing the number of considered possible front evolution directions. The performance of the technique is demonstrated in artificial data and in the healthy human brain. Comparisons with standard FM tractography and conventional line propagation algorithms show that, in the presence of interfering structures, the proposed method is more accurate in reconstructing trajectories. The in vivo results illustrate that the elucidated major white matter pathways are consistent with known anatomy and that multiple crossings and tract branching are handled correctly. PMID- 16249101 TI - Molecular phylogeny of the palm genus Chamaedorea, based on the low-copy nuclear genes PRK and RPB2. AB - Sequence data from the low-copy nuclear genes encoding phosphoribulokinase (PRK) and the second largest subunit of RNA polymerase II (RPB2) are used to generate the first phylogenetic analysis of Chamaedorea (Arecaceae: Arecoideae: Chamaedoreeae), the largest neotropical genus of palms. The prevailing current taxonomy of Chamaedorea recognizes approximately 100 species in eight subgenera, all delimited using floral characters, which provide a useful starting point to explore species-level systematics. Sequence data from 63 species, including representatives of all eight subgenera, were analyzed using maximum parsimony and Bayesian inference optimality criteria. Genus Chamaedorea is resolved as monophyletic with strong support in all separate and combined analyses. The less species-rich subgenera are convincingly monophyletic and can be diagnosed using morphological synapomorphies. In contrast, the two largest subgenera, Chamaedorea and Chamaedoropsis, which are supposedly distinguishable from each other by the degree of connation in the staminate petals, are both resolved as highly polyphyletic. Several well supported monophyletic groups resolved by these gene regions have never before been proposed within Chamaedorea and are challenging to delimit using morphological criteria. Although PRK proved more informative than RPB2, both regions have strong utility for interpreting species-level relationships among the palms, which are notoriously recalcitrant subjects for molecular phylogenetic studies. In addition, a paralog of the target copy of PRK identified during the analysis represents a potentially valuable source of phylogenetic information for future studies. PMID- 16249100 TI - Association of neural and physiological responses during voluntary emotion suppression. AB - Recent neuroimaging studies have shown that several prefrontal regions play critical roles in inhibiting activation of limbic regions during voluntary emotion regulation. The present study aimed to confirm prior findings and to extend them by identifying the frontal neural circuitry associated with regulation of peripheral physiological responses during voluntary emotion suppression. Ten healthy female subjects were presented with affectively positive, neutral, and negative pictures in each of an Attending and Suppression task. Regional cerebral blood-flow changes were measured using 15O-water positron emission tomography, and autonomic (heart rate: HR, skin conductance response: SCR) and endocrine (adrenocorticotropic hormone: ACTH) indices were measured during both tasks. The left amygdala and the right anterior temporal pole were activated during the Attending task, whereas activation was observed in the left lateral prefrontal cortex (LPFC), including the adjacent medial prefrontal cortex (MPFC), and medial orbitofrontal cortex (MOFC) during the Suppression task. In the Attending task, activation in the amygdala and MOFC positively correlated with magnitudes of the SCR and ACTH responses. Emotion suppression elicited enhancement of SCR and the strength of the effect positively correlated with activation in the MOFC. These results suggest that the MOFC plays a pivotal role in top-down regulation of peripheral physiological responses accompanying emotional experiences. PMID- 16249102 TI - Development of a time-resolved fluorometric assay for the high throughput screening of melanin concentrating hormone receptor antagonists. AB - INTRODUCTION: Melanin concentrating hormone is an orexigenic hypothalamic neuropeptide, which plays an important role in the complex regulation of energy balance and body weight mediated by the melanin concentrating hormone receptor subtype 1 (MCH1). Compelling pharmacological evidence implicating MCH1 signaling in the regulation of food intake and energy expenditure has generated a great deal of interest by pharmaceutical companies as MCH1 antagonists may have potential therapeutic benefit in the treatment of obesity and metabolic syndrome. METHODS: Although radioligand receptor binding assay has been one of the most powerful tools for receptor research and drug discovery, the limitations of radioisotopes and the problems related to safety and waste disposal limits their application in high throughput screening and has led to a growing interest in alternative, nonradioactive technologies. To develop a sensitive and reproducible assay system for MCH1, the time-resolved fluorescence (TRF) receptor binding assay with AcroWell filter plates was tested and validated. RESULTS: Comparing to the radioligand receptor binding assay for MCH1, the TRF assay presented higher Z/Z' factors with the lower signal-to-noise ratio. The known high-affinity MCH1 receptor antagonist, SNAP-7941, exhibited an IC50 value of 1.66+/-0.10 nM that is very similar to the IC50 value of MCH in a radioligand binding assay with an excellent correlation coefficient (0.9884). DISCUSSION: These results suggest that our TRF receptor binding assay for MCH1 can achieve the desired sensitivity and reproducibility to replace the radioligand receptor assay in a fluorometric system that can be developed for high throughput screening. PMID- 16249103 TI - Deamidation of peptides in aerobic nitric oxide solution by a nitrosative pathway. AB - Hydrolytic deamidation of asparagine (Asn) and glutamine (Gln) residues to aspartate (Asp) and glutamate (Glu), respectively, can have significant biological consequences. We hypothesize that a wholly different mechanism of deamidation might occur in the presence of aerobic nitric oxide (NO). Accordingly, we examined the deamidating ability of aerobic NO toward three model peptides, 2,4-dinitrophenyl (DNP)-Pro-Gln-Gly, Lys-Trp-Asp-Asn-Gln, and Ser-Glu Asn-Tyr-Pro-Ile-Val, incubating them with the NO-generating compound, Et(2)N[N(O)NO]Na (DEA/NO, 30-48 mM), in aerobic, pH 7.4, buffer at 37 degrees C. DNP-Pro-Glu-Gly was detected after 2 h, while Lys-Trp-Asp-Asp-Gln, Lys-Trp-Asp Asn-Glu, and Ser-Glu-Asp-Tyr-Pro-Ile-Val were detected within 10 min, accumulating in apparent yields of up to approximately 10%. In the latter case, tyrosine nitration was also observed, producing the expected nitrotyrosine residue. DEA/NO solutions preincubated to exhaust the NO before the peptides were added did not induce detectable deamidation. The data demonstrate that aerobic NO exposures can lead to nitrosative deamidation of peptides, a pathway that differs from the established hydrolytic deamidation mechanism in several key respects: the by-products of the former are molecular nitrogen and an acid (HONO) while that of the latter is a base (NH(3)); the nitrosative path can in principle proceed in the absence of water molecules; Asn is much more easily deamidated than Gln in the hydrolytic pathway, while the two amino acid residues were deamidated to a similar extent by exposure to NO in the presence of oxygen. PMID- 16249104 TI - Alteration of aortic function from streptozotocin-diabetic rats with Kilham's virus is associated with inducible nitric oxide synthase. AB - Kilham's rat virus (KRV) is a parvovirus commonly known to affect laboratory rats. Qualitative immunohistochemical analysis revealed that aorta isolated from KRV-infected streptozotocin (STZ)-induced diabetic adult rats expressed markedly greater levels of inducible nitric oxide synthase (iNOS) than aorta from KRV infected controls. In contrast with the prevailing literature, nitric oxide mediated endothelium-dependent relaxation to acetylcholine was not blunted by STZ diabetes, but was comparable to relaxations of aorta from controls. However, with increasing ex vivo duration, a decreased response to acetylcholine was observed in the STZ-diabetic aorta. In addition, whereas contraction responses to phenylephrine were not significantly altered over time in control tissue, aorta from STZ-diabetic rats developed increased tensions. The data suggest that increased iNOS-derived nitric oxide masks expected acetylcholine-mediated relaxation deficits as a result of KRV-infection, and that the deficit is unmasked by iNOS turnover ex vivo. PMID- 16249105 TI - Efficacy of oral vaccination against swine erysipelas in growing-finishing pigs in a clinically infected Slovakian pig herd. AB - In a large Slovakian growing-finishing pig production unit, the effects of oral vaccination against swine erysipelas (SE) were investigated in three groups of pigs of 10 weeks of age. In group 1, the pigs were vaccinated intramuscularly at 1 and 3 weeks after arrival in the growing-finishing barn using an Erysipelothrix rhusiopathiae bacterin. Group 2 pigs were vaccinated at the same time as group 1 using an oral avirulent live SE vaccine administered through drinking water; the pigs in the third group were placebo treated. Clinical signs of acute SE, arthritic changes, average daily weight gain (ADG), feed conversion ratio, and mortality were evaluated. None of the pigs in groups 1 and 2 but 31.7% of the control animals (group 3) showed typical clinical signs of acute SE. More (P<0.01) non-vaccinated pigs had chronic arthritic changes compared with groups 1 and 2. No significant differences in mortality were recorded between the groups. Groups 1 and 2 had higher (P<0.05) ADG and lower feed conversion ratios compared with group 3 pigs. The results demonstrated that the oral avirulent live culture was efficacious in significantly reducing the clinical symptoms caused by E. rhusiopathiae infection, so enhancing the pigs' performance. PMID- 16249106 TI - Comparison of Coccidioides immitis serological antibody titres between forms of clinical coccidioidomycosis in horses. AB - A retrospective study was performed to determine if there is an association between serological Coccidioides immitis antibody titres (IgG) and form/severity of coccidioidal disease in horses, and to identify trends in survival and treatment success based on the form of the disease. Data were obtained on horses with positive serological titres tested at the Coccidioidomycosis Serology Laboratory, School of Medicine, University of California, Davis from 1981 to 2004. Thirty-nine cases in which a diagnosis of coccidioidomycosis had been made were selected for inclusion. Six distinct categories were identified including abortion (n=6), miliary/interstitial pneumonia (n=6), pneumonia with thoracic effusion (pleural or pericardial) (n=11), disseminated (n=10), osteomyelitis (n=3) and external abscessation (n=3) both without pulmonary disease. Statistical differences in titre distribution were found between the abortion category and the pulmonary category (P=0.003), the abortion category and pneumonia with thoracic effusion (P=0.001), the abortion category and disseminated disease (P=0.001), and the pulmonary form and pneumonia with effusion (P=0.001). The other categories had overlapping titre results. Higher serological antibody titres seemed to be associated with a poorer prognosis for survival. Categories with the highest titres, disseminated (geometric mean titre=104) and pneumonia with thoracic effusion (geometric mean titre=226), were overwhelmingly fatal (19/21 known deaths) due to severe clinical disease. The categories with lower titres, abortion (geometric mean titre=4), bone involvement only (geometric mean titre=13) and cutaneous (geometric mean titre=5), had a better survival rate (10/12 known survivors) and less severe clinical disease. Measurement of serological titre may be a useful diagnostic aid in establishing form and severity of disease and thus inform prognosis. PMID- 16249107 TI - A monoclonal antibody against a canine CD45 homologue: analysis of tissue distribution, biochemical properties and in vitro immunological activity. AB - This report describes the characterisation of a monoclonal antibody (mAb), AB6, which recognises specifically a cluster of canine leukocyte surface molecules. The immunogen used for obtaining the AB6 mAb was a lysate of canine peripheral blood mononuclear cells (PBMC). This novel mAb belongs to the IgG2a isotype, and reacted in Western blot with four different canine leukocyte glycoproteins with apparent molecular weights of 180, 190, 205 and 220 kDa. The AB6 mAb recognised the majority of canine peripheral blood leukocytes as determined by flow cytometry (97%). It also exhibited a broad reactivity pattern against lymphoid and myeloid cells, inhibited the proliferation of mitogen-stimulated canine PBMC and did not recognise human PBMC and murine splenocytes. The biochemical properties, cell and tissue specificity, and in vitro biological activity of the AB6 mAb indicate that it recognises a canine CD45 homologue. The mAb could become a valuable diagnostic and research tool for the evaluation of immune functions in dogs. PMID- 16249108 TI - The challenge of assessing inflammatory and structural changes in lower equine airways: a chance for thoracoscopic-guided pulmonary biopsy? PMID- 16249109 TI - New opportunities for double rotation NMR of half-integer quadrupolar nuclei. AB - A combined approach is presented which expands the applicability of double rotation (DOR) by overcoming its most prominent disadvantages: spinning stability and sensitivity. A new design using air-bearings for the inner rotor and a computer-assisted start-up procedure allows DOR operation over in principle unlimited time at outer rotor speeds of up to 2000Hz. Sensitivity enhancement of the DOR experiment is achieved by applying amplitude-modulated adiabatic pulses such as the double frequency sweep (DFS) before pulse excitation. Repeating the DFS enhancement and signal readout several times without allowing for spin lattice relaxation leads to sensitivity enhancements of a factor 3 for (27)Al in various minerals. As a result, it becomes possible to study low sensitivity quadrupolar nuclei and various long duration 2D measurements can be performed routinely. Spinning is adequate to suppress residual homonuclear dipolar couplings in the spectral dimension of typical quadrupolar spin systems. In 2D exchange spectroscopy, however, homonuclear correlation can still be established through dipolar-quadrupolar cross-terms. PMID- 16249110 TI - Cooling overall spin temperature: protein NMR experiments optimized for longitudinal relaxation effects. AB - In experiments performed on protonated proteins at high fields, 80% of the NMR spectrometer time is spent waiting for the (1)H atoms to recover their polarization after recording the free induction decay. Selective excitation of a fraction of the protons in a large molecule has previously been shown to lead to faster longitudinal relaxation for the selected protons [K. Pervushin, B. Vogeli, A. Eletsky, Longitudinal (1)H relaxation optimization in TROSY NMR spectroscopy, J. Am. Chem. Soc. 124 (2002) 12898-12902; P. Schanda, B. Brutscher, Very fast two dimensional NMR spectroscopy for real-time investigation of dynamic events in proteins on the time scale of seconds, J. Am. Chem. Soc. 127 (2005) 8014-8015; H.S. Attreya, T. Szyperski, G-matrix Fourier transform NMR spectroscopy for complete protein resonance assignment, Proc. Natl. Acad. Sci. USA 101 (2004) 9642 9647]. The pool of non-selected protons acts as a "thermal bath" and spin diffusion processes ("flip-flop" transitions) channel the excess energy from the excited pool to the non-selected protons in regions of the molecule where other relaxation processes can dissipate the excess energy. We present here a sensitivity enhanced HSQC sequence (COST-HSQC), based on one selective E-BURP pulse, which can be used on protonated (15)N enriched proteins (with or without (13)C isotopic enrichment). This experiment is compared to a gradient sensitivity enhanced HSQC with a water flip-back pulse (the water flip-back pulse quenches the spin diffusion between (1)H(N) and (1)H(alpha) spins). This experiment is shown to have significant advantages in some circumstances. Some observed limitations, namely sample overheating with short recovery delays and complex longitudinal relaxation behaviour are discussed and analysed. PMID- 16249111 TI - Motional relativity and industrial NMR sensors. AB - We explore the implications of motional relativity in NMR and show that sample translation can be used to acquire NMR signals without the need for pulsed RF excitation or pulsed magnetic field gradients. Novel single-shot, on-line NMR acquisition protocols for samples being conveyed at high speed are discussed and preliminary results using a low-cost, on-line prototype NMR sensor are presented. PMID- 16249112 TI - Electrocardiogram and heart rate in response to temperature acclimation in three representative vertebrates. AB - Comparisons of electrocardiogram (ECG) and heart rate characteristics of three representative species in response to temperature acclimation were studied. In toad (Bufo raddei), T wave had positive, negative and flat patterns, which was different from positive in lizard (Eremias multiocellata), blunt and broad in bird (Alectories magna). The duration of P-R interval, Q-T interval and QRS complex interval reduced with increasing temperature in toad, but the P-R and T-P intervals were affected mostly, the QRS and R-T intervals were relatively less affected in lizard. In the bird, the voltage of P, S and T wave scarcely changed, R wave increased slightly with temperature going up in the thermal neutral zone (20-35 degrees C), T and S waves tended to increase and P-S and S-T intervals shortened when temperature went below the neutral zone. Heart rate was high and relatively steady in bird, but changed linearly in relation to temperature in toad and lizard. The increasing of heart rate with temperature was mainly caused by the T-P interval shortened in lizard, but P-S and S-T intervals shortened in bird. Comparisons of ECG and heart rate characteristics of three representative species in response to temperature acclimation reflected phylogenetically based constraints on pacemaker rates, oxygen supply and modulatory mechanisms. PMID- 16249113 TI - Oral cavity cancers at a young age: analysis of patient, tumor and treatment characteristics in Chiang Mai University Hospital. AB - Oral cavity cancer is predominantly a disease of middle-aged men who use tobacco and alcohol. Nearly 95% of carcinomas occur after the age of 45, with an average age of approximately 60 years. In recent years, oral cavity cancers have increased at a younger age, especially in females who never consumed alcohol or smoked. The purpose of this study is to provide the information of these cancers in young patients treated in our hospital during a 5-year period. As well as to describe the treatment modalities and their results. We reviewed the medical records of oral cavity cancer patients occurring before the age of 45 who were treated at Chiang Mai University Hospital from 1999 to 2003. All the demographic data, histopathology, treatment modalities and their results were recorded. Follow up range from 0.7 to 4.4 years (mean 2.6 year). A total of 20 patients were studied. There were 12 male (60%) and 8 female (40%). The mean age was 34.4 year (20-40 year). The most common site was oral tongue (15 patients, 75%). Fifty five percent of patients were stage III and IV. Only 6 patients (30%) were treated by surgery alone, 8 patients (40%) were treated by surgery and post operative radiotherapy, 4 patients (20%) were subjected to radiotherapy alone, and 2 patients (10%) were treated by radiochemotherapy. The results of primary treatment in all modalities were acceptable with 5 patients (25%) developing loco regional recurrence of disease within 10.8 months (2-36). At the time of analysis, 13 patients (65%) had no evidence of disease. The demographic data of oral cavity in younger patients in our hospital were different from the elderly, with oral tongue commonly occurring. Most of the patients were locally advanced stage. The results of all treatment modalities provided fair loco-regional control suggested more aggressive treatment in this group of patients. PMID- 16249114 TI - Independent prognostic factors of 861 cases of oral squamous cell carcinoma in Korean adults. AB - Oral squamous cell carcinoma (OSCC) accounts for 4.5% of all malignant tumors in Korean males and 3.5% in Korean females. The high recurrence rate, and in particular the high local recurrence rate, constitutes a major therapeutic problem for the Korean population, yet there is a paucity of reports addressing the independent predictors of response and survival rate of OSCC in Korea. The present study was designed to investigate the prognostic value of clinical and demographic data within a set of 861 cases of OSCC in Korea. The medical records of 861 OSCC patients who received treatment between 1984 and 1996 at 22 Korean hospitals were reviewed retrospectively with respect to several patient characteristics, including age at diagnosis, gender, location, TNM stage, and treatment. Independent patient-related and treatment-related factors that significantly influenced disease outcome after treatment were analyzed. To assess the independent factors affecting survival rate, univariate and multivariate regression analyses of the survival data were performed using the Cox proportional hazards model. A tree-structured survival model was also derived using survival tree with unbiased detection of interaction (STUDI). The multivariate Cox regression analysis showed that age, gender, composite stage, and treatment method were significant independent prognostic factors. Radiation dose, stage, size of tumor mass, and age of patient also strongly impacted survival time. OSCC is an extremely malignant carcinoma whose prognostic factors are multiple and complex. Based on the findings of this study, we believe that the prognosis of OSCC might depend directly on cancer stage as determined by the TNM system. Furthermore, the survival rate is positively affected by treatment of the neck upon presentation of the cancer, as this can prevent late neck disease due to persistent growth of occult metastases. PMID- 16249115 TI - Prognostic factors in head and neck adenoid cystic carcinoma. AB - The aim of this study was to analyse the clinical, histological and immunohistochemical prognostic factors of a large series of adenoid cystic carcinoma, using univariate and multivariate survival analyses. All cases of head and neck ACC (n = 129) treated in a single institution in Brazil, between 1955 and 1997, were selected for the study. Univariate survival analysis revealed that age older than 45 years (p = 0.04), period of complaints inferior to 18 months (p = 0.007), presence of paresthesia (p = 0.04), T stage (p = 0.01), N stage (p = 0.04), M stage (p < 0.001), clinical stage (p = 0.003), solid histological type ( p< 0.001), presence of residual tumor (p < 0.001) and expression of p53 (p = 0.08) correlated with a poor prognosis. In the multivariate survival analyses, clinical stage, solid histological subtype and increased expression of p53 were independent significant prognostic factors. According to our findings, clinical stage, solid growth pattern and expression of p53 were the most important prognostic factors in patients with ACC. PMID- 16249116 TI - Reduced E-cadherin expression is an indicator of unfavourable prognosis in oral squamous cell carcinoma. AB - The aim was to evaluate E-cadherin expression in oral squamous cell carcinoma, and its possible relationships with tumour histology and with clinical course and survival. Surgical biopsies from 47 cases of oral squamous cell carcinoma were analysed for expression of E-cadherin using immunohistochemistry. Statistical analyses were performed to identify possible associations with tumour clinic histological features and with clinical course and survival. Weak or absent E cadherin expression was associated with a more invasive histological pattern and with metastasis to the cervical lymph nodes. Uni- and multivariate analyses indicated that weak or undetectable E-cadherin expression is an indicator of shorter disease-free period and shorter survival time. Reduced E-cadherin expression in oral squamous cell carcinoma is associated with more aggressive tumour behaviour and worse prognosis. PMID- 16249117 TI - Interferon alpha2b augments suppressed immune functions in tobacco-related head and neck squamous cell carcinoma patients by modulating cytokine signaling. AB - We have examined the role of interferon alpha2b (IFNalpha2b) in augmentation of the suppressed immune functions and cytotoxicity of tobacco-related head and neck squamous cell carcinoma (HNSCC) patients. The suppressed killing activity of PBMC of HNSCC patients towards KB, MCF7 and K562 cell lines could be restored by in vitro treatment of PBMC with IFNalpha2b, as detected by LDH release assay. HNSCC patients with cisplatin + 5FU + IFNalpha2b treatment showed greater cytotoxic efficacy than corresponding pretreatment values. Analysis of culture supernatant of HNSCC-PBMC by ELISA revealed the lower secretion of IL-12 and IFNgamma with increased level of IL-4 and IL-10. This altered Th1/Th2 status was rectified after in vitro and in vivo IFNalpha2b stimulation. Increased secretion of monocyte derived IL-12 was observed after IFNalpha2b treatment that can enhance the IFNgamma release, a key regulator for cytotoxicity. IFNalpha2b stimulated enhancement of NK cells may be the source of greater amount of IFNgamma. IFNalpha2b activated STAT1 and STAT4 signaling is observed to be involved in the regulation and maintenance of cytokine milieu. We conclude that IFNalpha2b may be effective as a tool for adjuvant therapy along with conventional therapies to overcome the immunosuppression in HNSCC patients. PMID- 16249118 TI - Archaeal cell cycle progress. AB - The discovery of multiple chromosome replication origins in Sulfolobus species has added yet another eukaryotic trait to the archaea, and brought new levels of complexity to the cell cycle in terms of initiation of chromosome replication, replication termination and chromosome decatenation. Conserved repeated DNA elements--origin recognition boxes--have been identified in the origins of replication, and shown to bind the Orc1/Cdc6 proteins involved in cell cycle control. The origin recognition boxes aid in the identification and characterization of new origins, and their conservation suggests that most archaea have a similar replication initiation mechanism. Cell-cycle-dependent variation in Orc1/Cdc6 levels has been demonstrated, reminiscent of variations in cyclin levels during the eukaryotic cell cycle. Information about archaeal chromosome segregation is also accumulating, including the identification of a protein that binds to short regularly spaced repeats that might constitute centromere-like elements. In addition, studies of cell-cycle-specific gene expression have potential to reveal, in the near future, missing components in crenarchaeal chromosome replication, genome segregation and cell division. Together with an increased number of physiological and cytological investigations of the overall organization of the cell cycle, rapid progress of the archaeal cell cycle field is evident, and archaea, in particular Sulfolobus species, are emerging as simple and powerful models for the eukaryotic cell cycle. PMID- 16249119 TI - Determinants of transcription initiation by archaeal RNA polymerase. AB - Transcription in Archaea is catalyzed by an RNA polymerase that is most similar to eukaryotic RNA polymerases both in subunit composition and in transcription initiation factor requirements. Recent studies on archaeal transcription in diverse members of this domain have contributed new details concerning the functions of promoters and transcription factors in guiding initiation by RNA polymerase, and phylogenetic arguments have allowed modeling of archaeal transcription initiation complexes by comparison with recently described models of eukaryotic and bacterial transcription initiation complexes. Important new advances in reconstitution of archaeal transcription complexes from fully recombinant components is permitting testing of hypotheses derived from and informed by these structural models, and will help bring the study of archaeal transcription to the levels of understanding currently enjoyed by bacterial and eukaryotic RNA polymerase II transcription. PMID- 16249120 TI - Altered phase-transitions in tibialis anterior and medial gastrocnemius during walking after limbsaving surgery. AB - OBJECTIVE: In normal gait onset activity in tibialis anterior at end stance is closely linked to reduction in activity in medial gastrocnemius. Is a similar transition also present in patients undergoing limb-saving surgery? METHODS: Nineteen subjects after limb-saving surgery of the lower extremity and ten age matched controls were compared. Patients walked on a treadmill at their preferred speed. Bipolar surface EMG activity was recorded from the tibialis anterior and the medial gastrocnemius. RESULTS: Patients showed asymmetry in gait. In controls a close tibialis anterior and medial gastrocnemius connection was seen. The close link between ipsilateral tibialis anterior and medial gastrocnemius was absent in patients. Instead, a link was found between tibialis anterior onset in the affected and medial gastrocnemius onset in the non-affected leg. CONCLUSIONS: This finding suggests that two control mechanisms can be seen: a contralateral connection between tibialis and gastrocnemius and a less important ipsilateral connection. This means that automated phase switching in patients does not rely primarily on ipsilateral mechanisms but that instead the onset of the ipsilateral swing is linked to the moment of load acceptance by the contralateral leg These results are strikingly similar to those obtained in simulated limping by normal subjects. SIGNIFICANCE: Patients after limbsaving surgery have a clinically significant problem that creates aberrant gait patterns. This study provides new information about linking of ankle dorsiflexors and plantarflexors, as well as reporting the uncoupling of this mechanism in these patients after major surgery. PMID- 16249121 TI - Strain-specific mycobacterial lipids and the stimulation of protective immunity to tuberculosis. AB - Control of Mycobacterium tuberculosis (M. tuberculosis) infection is dependent on recognition of bacilli by cells of the innate immune system in the lung and the subsequent generation of an acquired effector T lymphocyte response. Lipid moieties of M. tuberculosis are important stimulators of innate immunity mediated predominantly through recognition by Toll-like receptors. In this paper, we will discuss how the lipid composition of different clinical isolates (strains) of M. tuberculosis affect that strain's ability to direct innate immunity, and ultimately influence whether infection is controlled or active disease develops. PMID- 16249122 TI - Animal models of tuberculosis. AB - It was Robert Koch who recognized the spectrum of pathology of tuberculosis (TB) in different animal species. The examination of clinical specimens from infected humans and animals confirmed the variable patterns of pathological reactions in different species. Guinea pigs are innately susceptible while humans, mice and rabbits show different level of resistance depending upon their genotype. The studies of TB in laboratory animals such as mice, rabbits and guinea pigs have significantly increased our understanding of the aetiology, virulence and pathogenesis of the disease. The introduction of less than five virulent organisms into guinea pigs by the respiratory route can produce lung lesions, bacteraemia and fatal diseases, which helped the extrapolation of results of such experiments to humans. The similarities in the course of clinical infection between guinea pigs and humans allow us to model different forms of TB and to evaluate the protective efficacy of candidate vaccines in such systems. The only limitation of this model, however, is a dearth of immunological reagents that are required for the qualitative and quantitative evaluation of the immune responses, with special reference to cytokines and cell phenotypes. Another limitation is the higher cost of guinea pigs compared with mice. The rabbit is relatively resistant to Mycobacterium tuberculosis, however following infection with virulent Mycobacterium bovis, the rabbit produces pulmonary cavities like humans. The rabbit model, however, is also limited by the lack of the immunological reagents. Mice are the animal of choice for studying the immunology of mycobacterial infections and have contributed much to our current understanding of the roles of various immunological mechanisms of resistance. The resistance of mice to the development of classic TB disease, however, represents a significant disadvantage of the mouse model. Although non-human primates are closely related to humans, owing to high cost and handing difficulties they have not been exploited to a large extent. As all existing animal models fail to mimic the human disease perfectly, efforts should be focused on the development of the non human primate(s) as the alternative animal model for TB. PMID- 16249123 TI - Bone loss in epilepsy: barriers to prevention, diagnosis, and treatment. AB - In epilepsy populations there are multiple barriers to the prevention, diagnosis, and treatment of metabolic bone disease. This study explored issues that may be unique to inner-city care. We screened 101 consecutive patient charts from our clinic to identify barriers to optimal care. According to the Health Belief Model, when patients view a condition as not severe and their perceived susceptibility is low, they are unlikely to comply with screening and/or adopt preventative behaviors. Despite routine communications to primary care doctors regarding bone health, its importance is still overlooked. In addition, the managed care referral process makes it difficult for a specialist to prevent, screen, and treat a primary care condition with no symptoms. For optimal patient care and to stimulate prevention and screening compliance, a communication style adapted to the patient's knowledge, beliefs, and attitudes toward antiepileptic drug-induced bone loss is crucial to help reduce barriers against change. PMID- 16249124 TI - Catatonia induced by levetiracetam. AB - Levetiracetam (Keppra) is a novel antiepileptic drug approved as adjunctive treatment for adults with partial onset seizures. Although the drug is generally well tolerated, behavioral side effects have been reported in variable frequency. Most behavioral problems are mild in nature (agitation, hostility, anxiety, emotional lability, apathy, depression) and quickly resolve with discontinuation of medication. However, serious psychiatric adverse events may also occur with rare cases of psychosis and suicidal behavior. We report here the case of a 43 year-old woman who developed symptoms compatible with catatonia after being exposed to levetiracetam for the treatment of epilepsy. To our knowledge, it is the first reported case of catatonia induced by levetiracetam. We review the difficulties that may be encountered in the differential diagnosis of medical catatonia. PMID- 16249125 TI - Expression profiling in the aging brain: a perspective. AB - To evaluate molecular events associated with the aging process in animal models and human tissues, microarray analysis is performed at the regional and cellular levels to define transcriptional patterns or mosaics that may lead to better understanding of the mechanism(s) that drive senescence. In this review, we outline the experimental and analytical issues associated with high-throughput genomic analyses in aging brain and other tissues for a comprehensive evaluation of the current state of microarray analysis in aging paradigms. Ultimately, the goal of these studies is to apply functional genomics and proteomics approaches to aging research to develop new tools to assess age in cell- and tissue-specific manners in order to develop aging biomarkers for pharmacotherapeutic interventions and disease prevention. PMID- 16249126 TI - Liquid chromatography-tandem mass spectrometry (LC-MS-MS) method for simultaneous determination of venlafaxine and its active metabolite O-desmethyl venlafaxine in human plasma. AB - A rapid and sensitive liquid chromatography-tandem mass spectrometry (LC-MS-MS) method has been developed and validated for simultaneous quantification of venlafaxine (VEN) and O-desmethyl venlafaxine (ODV) in human plasma. The analytes were extracted from human plasma by using solid-phase extraction (SPE) technique. Escitalopram (ESC) was used as the internal standard. A Betasil C18 column provided chromatographic separation of analytes followed by detection with mass spectrometry. The mass transition ion-pair has been followed as m/z 278.27- >121.11 for VEN, m/z 264.28-->107.10 for ODV and m/z 325.00-->262.00 for ESC. The method involves a solid phase extraction from plasma, simple isocratic chromatography conditions and mass spectrometric detection that enables detection at nanogram levels. The proposed method has been validated with linear range of 3 300 ng/ml for VEN and 6-600 ng/ml for ODV. The intrarun and interrun precision and accuracy values are within 10%. The overall recoveries for VEN and ODV were 95.9 and 81.7%, respectively. Total elution time as low as 3 min only. PMID- 16249127 TI - Crystal structures of the free and sterol-bound forms of beta-cinnamomin. AB - The crystal structure of the elicitin beta-cinnamomin (beta-CIN) was determined in complex with ergosterol at 1.1 A resolution. beta-CIN/ergosterol complex crystallized in the monoclinic space group P2(1), with unit cell parameters of a = 31.0, b = 62.8, c = 50.0 A and beta = 93.4 degrees and two molecules in the asymmetric unit. Ligand extraction with chloroform followed by crystallographic analysis yielded a 1.35 A structure of beta-CIN (P4(3)2(1)2 space group) where the characteristic elicitin fold was kept. After incubation with cholesterol, a new complex structure was obtained, showing that the protein retains, after the extraction procedure, its ability to complex sterols. The necrotic effect of beta CIN on tobacco was also shown to remain unchanged. Theoretical docking studies of the triterpene lupeol to beta-CIN provided an explanation for the apparent inability of beta-CIN to bind this ligand, as observed experimentally. PMID- 16249128 TI - Spontaneous regression of hepatitis B virus-associated cirrhosis developed in childhood. AB - We reported two cases of hepatitis B virus infection-related cirrhosis developed during childhood and followed up for more than 20 years. Both the subjects remained untreated, and ultimately regression of cirrhosis was documented by clinical (including ultrasound) and histological examination. Recent studies have already suggested that hepatitis B virus-related cirrhosis may regress after treatment, but this is the first demonstration that hepatitis B virus-associated cirrhosis developed in childhood may be a spontaneously reversible process. Subsidence of virus replication and of necro-inflammatory process and the efficiency of liver regeneration and repair might contribute to this favourable outcome. PMID- 16249129 TI - Mast cell activation and degranulation occur early during induction of periosteal bone resorption. AB - We have previously postulated that mast cells participate in the cellular network involved in osteoclastic resorption, probably through histamine release. In this study, we examined mast cell activation and histamine release during origination of resorption. Groups of 10 rats were killed 0, 0.5, 1, 1.5, 3, 6, 9, 12 and 18 h after induction of resorption in a synchronized model of cortical resorption along the mandible. The total number of mast cells was transiently decreased by about one-third at 1 and 9 h. Mast cell activation was monitored by Alcian blue safranin staining. Early after induction, mast cells started to release their mediator stores; complete release led to the apparent disappearance of the cells with the staining technique used. Histamine immunostaining confirmed the release of histamine and its diffusion in the extracellular environment. Massive degranulation was observed at 1.5 and 9 h with toluidine blue staining. Cell recovery, assessed in terms of histidine decarboxylase expression, occurred gradually. The number of ED1+ osteoclast precursors strongly increased from 12 h up to 18 h. Most parameters had returned to baseline at 18 h, except the ED1+ cells. H2 receptor inhibition with famotidine strongly decreased ED1+ osteoclast precursors at 12 h and subsequently osteoclasts at the peak of resorption. These data support a role of mast cells in resorption origination. They show an early and transient intervention of mast cells in the events regulating the recruitment of circulating osteoclast precursors and ultimately of resorption. Mast cell activation and degranulation induce the release of mediators, particularly histamine acting through its H2 receptors, which are likely involved in these reactions. PMID- 16249130 TI - Low width of tubular bones is associated with increased risk of fragility fracture in elderly men--the MINOS study. AB - The risk of fragility fractures in elderly men is only partly explained by areal bone mineral density (aBMD) measured by dual X-ray absorptiometry (DXA). Several studies suggest the importance of bone morphology for the risk of fracture. The aim of this study was to assess the value of bone size and estimated structural parameters for the prediction of incident fractures in a large cohort of men. This study was made in 759 men aged 50-85 from the MINOS cohort. During a 90 month follow-up, 74 men sustained incident vertebral and peripheral fractures. Areal BMD was measured by DXA at femoral neck, distal radius and distal ulna. Estimates of structural bone parameters and volumetric BMD (vBMD) were derived from aBMD measured by DXA. Given the limited number of fractures, the predictive value of investigated parameters was assessed for peripheral and vertebral fractures jointly by using logistic regression. Men who sustained the fractures had, at baseline, lower aBMD (3.5-6.5%), lower bone mineral content (BMC 5.4 8.7%) and lower cortical thickness (3.5-6.9%) compared with the men without fracture. At all the three skeletal sites, aBMD, BMC, width, cortical area and thickness, cross-sectional moment of inertia (CSMI), and section modulus predicted incident fractures (O.R. = 1.28-1.92 per 1 SD decrease, P < 0.05 0.0001). Fracture risk was weakly associated with vBMD for ulna (O.R. = 1.25 per 1 SD decrease, P < 0.05) but not for femoral neck or radius. After adjustment for aBMD, bone width remained a significant predictor of fractures (O.R. = 1.37-1.48 per 1 SD decrease, P < 0.02-0.01). Men with osteopenia (BMD T score < -1) and low bone width (T score < -1) had the fracture incidence similar to that observed in men with BMD T score < -2. Bone width and aBMD of the femoral neck and radius were predictive of fractures in 49 men with the incident peripheral fractures, whereas their O.R. did not attain the level of statistical significance in 25 men with the incident vertebral fractures. Men, who had both low aBMD and low CSMI ( both T scores < -1), had the fracture risk 3.8 to 4.2 higher than the reference group (both T scores >or= -1). Men, who had both low aBMD and low section modulus (both T scores < -1), had the fracture risk 2.1 to 4.1 higher than the reference group (both T scores >or= -1). In conclusion, men who sustained a fragility fracture during a 90-month follow-up had, at baseline, lower BMC because they had narrower bones but not necessarily less dense. In elderly men, small bone width, low BMC and poor resistance to bending may increase bone fragility. Low bone width seems to be associated with an increased fracture risk in elderly men regardless of aBMD. PMID- 16249131 TI - Exclusion mapping of chromosomes 1, 4, 6 and 14 with bone mineral density in 79 Caucasian pedigrees. AB - Low bone mineral density (BMD) is a major determinant of osteoporosis and is under strong genetic control. A large number of linkage and association studies for BMD variation have been conducted, with the results being largely inconsistent. Linkage exclusion analysis is a useful tool for gene mapping but has never been used on BMD. In the present study, we conducted a linkage exclusion mapping for BMD variation on chromosomes 1, 4, 6 and 17 in 79 Caucasian pedigrees. For hip BMD variation, several genomic regions were excluded for effect sizes of 10% or greater, including regions of 61-77 cM at 1p35-p34, 167 196 cM at 1q21-q23 and 261-291 cM at 1q42-q44; 85-112 cM at 4q21-q25 and 146-150 cM at 4q31; and 77-85 cM at 6p12-q13. For spine BMD, we were able to exclude the regions of 168-189 cM at 1q21-q23, 92-94 cM at 4q21 and 106-107 cM at 4q24 and 56 103 cM at 17q12-q25, as having effect sizes of 10% or greater. These results suggest that a number of candidate genes located in the excluded regions, such as interleukin 6 receptor (IL6R) gene, type I collagen alpha 1 (COL1A1) gene and bone morphogenetic protein-3 (BMP3) gene are unlikely to have a substantial effect on BMD variation in this Caucasian population. Along with previous studies searching for genes underlying BMD variation, the current study has further delineated the genetic basis of BMD variation and provided valuable information for future genetic studies. PMID- 16249136 TI - "Is there life on dialysis?": time and aging in a clinically sustained existence. AB - Increasingly, in the United States, lives are being extended at ever-older ages through the implementation of routine medical procedures such as renal dialysis. This paper discusses the lives and experiences of a number of individuals 70 years of age and older at two dialysis units in California. It considers what kind of life it is that is being sustained and prolonged in these units, the meanings of the time gained through (and lost to) dialysis for older people, and the relationship of "normal" life outside the units to an exceptional state on the inside that some patients see as not-quite-life. Highlighting the unique dimensions of gerontological time on chronic life support, the article offers a phenomenology of the end of life as that end is drawn out, deferred by technological means, and effaced by the ethos and experiential course of dialysis treatment. PMID- 16249132 TI - Inulin and fructo-oligosaccharides differ in their ability to enhance the density of cancellous and cortical bone in the axial and peripheral skeleton of growing rats. AB - Although dietary fructans improve calcium absorption and bone mineral content (BMC) in rats, their effect on calcium bioavailability and bone density may vary with their degree of polymerization. Therefore, for a 3-month period, growing rats received either a control diet or a diet enriched with either oligofructose (OLF) or inulin. At sacrifice, body weight, lean body mass and appendicular bone length were similar in the 3 groups. Rats fed fructans had a similar increase in cecal wall weight (30%), but the relative increase in cecal levels of calbinding 9 K was 2 in the OLF group and 4 in the inulin group. Further, the significant decrease in serum levels of type I collagen C-telopeptide was greater in the inulin group (30%) than in the OLF group (16%). The increase in whole-body bone mineral content (BMC) as measured by DXA was greater in the inulin group than in the OLF group but DXA detected an increase in the BMC of excised femurs only in the inulin group. In contrast, pQCT conducted ex vivo detected a significant increase in the area and mineral density (BMD) of the cancellous bone of both the proximal tibia and vertebra in rats fed fructans and the effect of inulin was greater (P < 0.01) than that of OLF (P < 0.05). Further, inulin but not OLF significantly enhanced the BMD of the cortical bone in both appendicular and peripheral sites (P < 0.01) as well as the polar stress/strain index of femurs (P < 0.01). These observations suggest that, although both inulin and OLF both have a positive effect on BMD, the greatest effect of inulin is related to the higher capacity of this fructan to reduce bone resorption. The different anti-resorptive capacity of the 2 fructans might be related to their different impact on calcium absorption and bioavailability since the increase in cecal amounts of calbindin-9 K, a protein known to play an important role in calcium absorption, was greater in rat fed inulin than in rats fed OLF. Although cecal wall hyperplasia may be of concern, it remains to establish whether the positive effect of fructans observed on calcium absorption in humans is also associated with a positive effect on bone mass and/or mineral density. PMID- 16249137 TI - Holistic physicians' clinical discourse on risk: an ethnographic study. AB - As physicians integrate complementary and alternative therapies into medical practice, they are compelled to develop new educational strategies for their patients. Therapeutic modalities whose success depends heavily upon significant behavior modification in patients require that physicians orient the latter to reconsider relationships between disease, environment, risk, and individual responsibility. This research explores the language physicians select to communicate to patients their perspectives on disease etiology and treatment as well as the importance of patients taking responsibility for changing their behavior. My data are based on fourteen months of research in a holistic medicine clinic, and they include physician interviews and observations of clinical encounters with patients. Holistic physicians often focus on diet as a major component of heath and disease, and they frequently make use of culturally relevant conceptions of risk related to food quality and environment in order to persuade patients to engage in prescribed behavior modification. PMID- 16249138 TI - Games of truth: rethinking conformity and resistance in narratives of heroin recovery. AB - Conventional paradigms of drug treatment present addicts as liars, fabricators, and manipulators until they are truly involved in their recovery process. My ethnographic study engages with and expands upon Foucault's (1988) concept of "games of truth" and De Certeau's (1984) work on "strategies" and "tactics" to illuminate complex and shifting relations between and among staff and clients in a methadone clinic in Toronto, Canada. I suggest the notion of "complicity" in order to address radical instability in these relationships, and I also question commonly held binaries, such as truth and lies, enunciation and action, domination and resistance, that underlie theoretical and addiction treatment realms. In this process, I contribute not only to anthropological theorizing but also to policy making. The staff and clients' multifaceted interpretations of heroin recovery question a single standard of "successful" treatment and point to the need for broader social interventions to address the diverse needs of heroin users. PMID- 16249139 TI - Does the teratogenicity of isotretinoin outweigh its benefits? PMID- 16249140 TI - Management of leukocytoclastic vasculitis. AB - Leukocytoclastic vasculitis (LcV) is the most common form of vasculitis of the skin and usually results from deposition of immune complexes at the vessel wall. It presents in different forms and in association with different diseases. When IgA is the dominant immunoglobulin in immune complexes, systemic involvement is likely in both children and adults (Henoch-Schonlein purpura--HSP). LcV due to IgG- or IgM-containing immune complexes has less systemic involvement and a better prognosis than HSP. Other forms of LcV include cryoglobulinaemic, urticarial and ANCA-associated LcV as well as LcV associated with vasculopathy and coagulopathy in SCLE/SLE or in bacteraemia/sepsis. The aim of diagnostic guidelines is to determine the specific type and systemic involvement of LcV and to identify an underlying cause. Basic work-up should encompass history of drug intake and of preceding infections, biopsy with immunofluorescence, differential blood count, urine analysis and throat swabs. Therapy of immune complex LcV often does not require aggressive therapy due to a usually favourable course. It includes avoidance or treatment of eliciting agents and use of compression stockings to reduce purpura. There are no large prospective randomized controlled studies. Corticosteroids are indicated when there are signs of incipient skin necrosis. In chronic or relapsing LcV we suggest colchicine as a first-line and dapsone as a second-line therapy. Corticosteroids may reduce the incidence of severe renal insufficiency in children according to some studies, but there is no study showing such an effect in adults. Severe systemic vasculitis requires immunosuppressive strategies. PMID- 16249141 TI - Tigecycline: a review of a new glycylcycline antibiotic. AB - Tigecycline derived from minocycline. It is part of a new class of antibiotics called glycylcyclines. Tigecycline is given intravenously and has activity against a variety of gram-positive and gram-negative bacterial pathogens, many of which are resistant to existing antibiotics. Tigecycline successfully completed phase III trials in which it was at least equal to intravenous vancomycin and aztreonam to treat complicated skin and skin structure infections (cSSSI), and to intravenous imipenem and cilastatin to treat complicated intra-abdominal infections (cIAI). Tigecycline side effects are primarily digestive upset. It should be a valuable addition to the armamentarium to treat even the most resistant pathogens. PMID- 16249142 TI - An open study to determine the efficacy of blue light in the treatment of mild to moderate acne. AB - BACKGROUND: The effective management of acne remains a challenge; achieving an optimal response whilst minimizing adverse events is often difficult. The rise in antibiotic resistance threatens to reduce the future usefulness of the current mainstay of therapy. The need for alternative therapies remains important. Phototherapy has previously been shown to be effective in acne, with renewed interest as both endogenous and exogenous photodynamic therapies are demonstrated for this condition. OBJECTIVES: To determine the effect of narrowband blue light in the reduction of inflammatory and non-inflammatory lesions in patients with mild to moderate acne and to evaluate patient tolerance of the therapy. METHODS: We performed an open study utilizing a blue LED light source in 30 subjects with mild to moderate facial acne. Two weeks after screening, lesions were counted and recorded by lesion type. Over 4 weeks, patients received eight 10- or 20-minute light treatments, peak wavelength 409-419 nm at 40 mW/cm2. Assessments were taken at weeks 5, 8 and 12 and lesion counts were recorded. Repeated measures-ANOVA and Dunnett's tests, respectively, allowed assessment of the different scores over time and permitted comparison of mean counts. RESULTS: An overall effect on inflammatory counts was observed at week 5, and a statistically significant decrease in inflamed counts was detected at the week 8 assessments, which continued to week 12. There was little effect on non-inflamed lesions. The treatment was well tolerated with adverse events experienced generally rated as being mild and usually self-limiting. CONCLUSIONS: Eight 10- or 20-minute treatments over 4 weeks with a narrowband blue light was found to be effective in reducing the number of inflamed lesions in subjects with mild to moderate acne. The treatment had little effect on the number of comedones. The onset of the effect was observable at the first assessment, at week 5, and maximal between weeks 8 and 12. Blue light phototherapy using a narrowband LED light source appears to be a safe and effective additional therapy for mild to moderate acne. PMID- 16249143 TI - Trends in prescription of acne medication in the US: shift from antibiotic to non antibiotic treatment. AB - AIM: To describe trends in the prescription of medication for acne vulgaris in the USA from 1990 to 2002 with particular reference to the shift from antibiotic to non-antibiotic treatment. METHODS: Retrospective cross-sectional observational study which analysed the data from the 4922 patient visits for acne vulgaris from the 1990-2002 National Ambulatory Medical Care Survey (NAMCS) to assess medications mentioned or prescribed at office visits for acne vulgaris. We specifically analysed visits for benzoyl peroxide or combination benzoyl peroxide products, topical retinoids, clindamycin, erythromycin, tetracycline group antibiotics and isotretinoin. A priori predictions were made for changes in physician behaviour over time, and were tested by regression. Multivariate regression also assessed the impact of demographic variables such as patient age, gender, race and type of physician seen on prescribing behaviour. RESULTS: From 1990 to 2002 there were significant declines (p<0.01) in the likelihood of use of several drug classes that are reliant on antimicrobial mechanisms for acne including the following: benzoyl peroxide, topical clindamycin, oral erythromycin and tetracycline group antibiotics. There were significant increases in likelihood of utilization for agents not reliant on antimicrobial mechanisms including topical retinoids and oral isotretinoin during this period. These changes in utilization were not affected by controlling for demographic factors in multivariate analysis. CONCLUSION: There has been a shift towards non antibiotic treatment in acne management. Growing awareness of antibiotic resistant Propionibacterium species may contribute to increased use of topical and systemic retinoid agents. PMID- 16249144 TI - An analysis of nursing activity in a day care and inpatient dermatology service. AB - The nursing context within which dermatological treatments are provided has received limited investigation. This activity analysis survey examined how nursing time is organized (n=17 nurses) within in- and out-patient day care services in a hospital teaching dermatology department, using a newly developed standardized tool over a 24-week period. The setting was a day care and inpatient facility in a UK dermatology department. Patients (n=108) referred to these services for day care or inpatient care over 24 weeks, excluding emergencies, were recruited consecutively; 76% (n=82) were seen during this time. Most patients (>95%) were referred for clearance of an acute episode of a chronic skin condition and specifically education. The total mean time in contact with the nursing service (mean hours/case) was 43.1 for inpatient and 6.6 for day care. Time in day care was mainly spent (mean hours/case) on direct care (living activities and dermatological treatments) (4.3), followed by teaching (2) and support (1.6). The pattern of inpatient care was similar. A profile has been derived that is indicative of how dermatology nurses spend their time in day and inpatient care. There may be scope for day care nursing to make more effective use of education opportunities to support patient self-management. PMID- 16249145 TI - Evaluation of the efficacy of topical caffeine in the treatment of psoriasis vulgaris. AB - BACKGROUND: Psoriasis is a common disease which often requires long-term maintenance therapy. In psoriatic epidermis, the level of cyclic adenosine monophosphate (cAMP) decreases. It has been reported that beta-blockers exacerbate existing psoriatic plaque and decrease the concentration of intracellular cAMP. Caffeine is a methylxanthine that inhibits phosphodiesterase enzyme and results in a higher concentration of intracellular cAMP. OBJECTIVE: Evaluation of the efficacy of topical caffeine 10% in the treatment of psoriasis. PATIENTS AND METHODS: The patients were treated by topical application of 10% caffeine or placebo three times per day on the right or left side of the body (randomly selected by flipping a coin). Thirty-nine patients with stable plaque psoriasis were included in a randomized, double-blind, placebo-controlled, right/left comparison. The patients visited every other week for a period of 8 weeks. Their Psoriatic Area and Severity Index (PASI) scores were assessed at each visit. RESULTS: The reductions in PASI scores measured at the four visits for the caffeine-treated group were 2.64+/-2.89, 4.47+/-3.62, 5.73+/-4.16, 6.58+/ 4.40 and for the placebo-treated group the values were 1.45+/-2.32, 3.04+/-2.68, 4.02+/-3.36, 4.43+/-3.45, respectively. Comparing the corresponding results of the two groups, p values at the second, fourth, sixth and eighth week were 0.081, 0.083, 0.079 and 0.047, respectively. Based on presented p values, the treatment with caffeine is more effective than with placebo after 8 weeks (p<0.05), and the only side effect of caffeine is mild itching. CONCLUSION: Based on the results of the trial, topical caffeine is an effective, safe and inexpensive treatment for psoriasis, with a delay in action. PMID- 16249146 TI - Clinical experience with the use of cyclosporin A in psoriasis. Results of a retrospective study. AB - The efficacy of cyclosporin A (Sandimmun Neoral) in severe psoriasis was analysed in 70 patients included in a retrospective descriptive study conducted at our Dermatology Department from January 1994 until December 2000. After 8 weeks of treatment, with an initial dose of 2.5-3 mg/kg/day, the reduction in the Psoriasis Area and Severity Index (PASI) score was>75% in 61% of patients. Most of the side effects observed turned out to be susceptible to being reversed over time or by dose reduction. The results of this study show that continuous and low dose cyclosporin therapy is a safe and efficacious treatment for severe chronic psoriasis. PMID- 16249147 TI - Treatment of Kimura's disease with cyclosporine. AB - Kimura's disease is a rare, chronic inflammatory disease of unknown cause. It is characterized by painless subcutaneous swellings and lymphadenopathy, commonly affecting the head and neck region. This is associated with peripheral blood eosinophilia and raised serum IgE. It has distinct histological features of lymphoid follicles, eosinophilic infiltrate, fibrosis and vascular proliferation. The disease usually has a benign, indolent course. Traditionally, therapeutic options have included surgery, radiotherapy and steroids but response has been less than satisfactory. Recently, cyclosporine has been reported to be effective in the treatment of Kimura's disease. In this article, we present a middle-aged Chinese female with Kimura's disease for 20 years and her favourable response to cyclosporine. PMID- 16249148 TI - A case of chronic actinic dermatitis treated with topical tacrolimus. AB - INTRODUCTION: The treatment of chronic actinic dermatitis (CAD), a sun-induced disorder characterized by a persistent eczematous eruption, involves photoprotective measures, topical corticosteroid therapy and, in more severe cases, systemic immunosuppression. The potential side effects of systemic immunosuppressant drugs prompted us to evaluate the efficacy of topical tacrolimus for treatment of CAD. PATIENT AND TREATMENT: A 58-year-old man with CAD, resistant to previous treatment with topical and systemic corticosteroids, oral cyclosporine and PUVA-photochemotherapy, was treated with tacrolimus ointment 0.1% once a day. RESULTS: Tacrolimus ointment led to significant improvement of pruritus and severe eczematous skin lesions after 20 days of treatment. CONCLUSIONS: Tacrolimus shows a beneficial effect on CAD; this could be attributed to the fact that CAD is characterized by a lymphohistiocytic infiltrate producing a chronic eczema and that tacrolimus blocks the activation of lymphocytes and other immune system cells, also inhibiting the release of mediators from cutaneous mast cells and basophils. PMID- 16249150 TI - The momentum of mortality change. AB - Mortality change is not usually assigned much importance as a source of population growth when future population trends are discussed. Yet it can make a significant contribution to population momentum. In populations that have experienced mortality change, cohort survivorship will continue varying for some time even if period mortality rates become constant. This continuing change in cohort survivorship can create a significant degree of mortality-induced population change, a process we call the 'momentum of mortality change'. The momentum of mortality change can be estimated by taking the ratio of e0 (the period life expectancy at birth) to CAL (the cross-sectional average length of life) for a given year. In industrialized nations, the momentum of mortality change can attenuate the negative effect on population growth of declining fertility or sustained below-replacement fertility. In India, where population momentum has a value of 1.436, the momentum of mortality change is the greatest contributor to its value. PMID- 16249149 TI - Social networks, perceptions of risk, and changing attitudes towards HIV/AIDS: new evidence from a longitudinal study using fixed-effects analysis. AB - The study presented here is an investigation of the importance of social interactions to perceptions of the risk of AIDS, and explores spousal communication about the AIDS epidemic in rural Malawi. A fixed-effects analysis based on longitudinal data collected in 1998 and 2001 shows that social interactions on the subject of HIV/AIDS have significant and substantial effects on respondents' perceptions of the risk of HIV/AIDS, even after controlling for unobserved factors that affect the selection of social networks. These effects are more complex than previously thought. The dominant mechanisms--social learning and social influence--are found to vary by sex and by region, because of regional variations in the marriage pattern and the resulting implications for the formation of local social networks. The conclusion of the study is that rather than fostering denial and inaction, social interactions are an important vector of change in the face of the HIV/AIDS epidemic. PMID- 16249151 TI - Older adults in sub-Saharan Africa living with children and grandchildren. AB - Using data from Demographic and Health Surveys, we examine the composition of households containing older adults in 24 countries of sub-Saharan Africa, with a focus on those living with children and grandchildren. Overall, 59 per cent live with a child and 46 per cent with a grandchild. Men are more likely to live in nuclear households and women in extended households and alone. Regression analyses show that individual-level determinants of household composition differ by sex. For example, living with children and grandchildren is tied to living with a spouse for men, but for women the effect is either not significant or in the opposite direction. Households with an older adult and a grandchild, but no adult children, are common. Usually the adult child lives elsewhere, though about 8 per cent of older adults live with a grandchild who has at least one deceased parent. Older adults are more likely to be living with double-orphans in countries with high AIDS-related mortality. PMID- 16249152 TI - Mortality inequality in two native population groups. AB - A sample of people aged 40-67 years, taken from a longitudinal register compiled by Statistics Finland, is used to analyse mortality differences between Swedish speakers and Finnish speakers in Finland. Finnish speakers are known to have higher death rates than Swedish speakers. The purpose is to explore whether labour-market experience and partnership status, treated as proxies for measures of variation in health-related characteristics, are related to the mortality differential. Persons who are single, disability pensioners, and those having experienced unemployment are found to have substantially higher death rates than those with a partner and employed persons. Swedish speakers have a more favourable distribution on both variables, which thus notably helps to reduce the Finnish-Swedish mortality gradient. A conclusion from this study is that future analyses on the topic should focus on mechanisms that bring a greater proportion of Finnish speakers into the groups with poor health or supposed unhealthy behaviour. PMID- 16249153 TI - High-risk families: the unequal distribution of infant mortality in nineteenth century Sweden. AB - An analysis of infant mortality (based on 133,448 births) in two regions, Sundsvall and Skelleftea, in north-eastern Sweden during the nineteenth century shows that infant mortality was highly clustered with a relatively small number of families accounting for a large proportion of all infant deaths. Using logistic regression, two important factors were found to be associated with high risk families: (i) a biological component evidenced by an over-representation of women who had experienced stillbirths, and (ii) a social component indicated by an increased risk among women who had remarried. The results strengthen the argument for using the family rather than the single child as the unit of analysis. The clustering of infant deaths points to the need to re-evaluate our interpretations of the causes of infant mortality in the past. PMID- 16249154 TI - The effect of non-family experiences on age of marriage in a setting of rapid social change. AB - This study examines the changing effects of non-family activities on the age of transition to first marriage in four cohorts of individuals across 45 years in the Chitwan Valley, Nepal. The results indicate that school enrolment had a negative effect on both men's and women's marriage rates, while total years of schooling had a positive effect on men's marriage rates. Non-family employment experiences increased marriage rates for men only. Analysing the effects of schooling and employment over time suggests that school enrolment became a growing deterrent to marriage for both sexes, and that non-family employment became an increasingly desirable attribute in men. The results are consistent with changing views about sex roles and schooling over time in the region, as the roles of student and spouse became more distinct. The results also suggest an increasing integration of husbands in the non-family labour market. PMID- 16249155 TI - Fertility in Kenya and Uganda: a comparative study of trends and determinants. AB - Between 1980 and 2000 total fertility in Kenya fell by about 40 per cent, from some eight births per woman to around five. During the same period, fertility in Uganda declined by less than 10 per cent. An analysis of the proximate determinants shows that the difference was due primarily to greater contraceptive use in Kenya, though in Uganda there was also a reduction in pathological sterility. The Demographic and Health Surveys show that women in Kenya wanted fewer children than those in Uganda, but that in Uganda there was also a greater unmet need for contraception. We suggest that these differences may be attributed, in part at least, first, to the divergent paths of economic development followed by the two countries after Independence; and, second, to the Kenya Government's active promotion of family planning through the health services, which the Uganda Government did not promote until 1995. PMID- 16249158 TI - Hip pain in adolescents: results of a cross-sectional study in German pupils and a review of the literature. AB - AIM: To determine the prevalence of hip pain (HP) in adolescents in East Germany and to evaluate the potential factors associated with HP. DESIGN: Cross-sectional epidemiological study of a total of 2368 pupils (1137 males, 1231 females; age 14.5+/-0.65, range 13-18 y). Demographic data and information on sports activity and consumption of soft legal drugs were obtained using a questionnaire. Physical examination was carried out by school doctors. Odds ratio (OR [95% confidence interval]) was calculated for common factors associated with HP. RESULTS: The prevalence of adolescents suffering from HP was 6.5% (n=151). Pain episodes were reported as occasional in 3.0%. A total of 69 subjects (2.9%) suffered from permanent pain during physical activity. Permanent pain when at rest was reported in 0.5% (n=12). Female subjects, students attending high school, smoking adolescents and adolescents with a consumption of alcoholic beverages had a significantly higher prevalence of HP. HP was associated with a significantly higher prevalence of low back pain and knee pain. The prevalence of pathological findings of the hip was 2.5% (n=60). Factors significantly associated with HP were: female gender (1.9-fold), attending high school (2.1-fold) and consumption of tobacco (1.6-fold). CONCLUSIONS: HP in adolescents seems to be mostly caused by functional disorders. There is a possible association between HP and psychological stress or consumption of legal drugs. Prophylactic measures are necessary in this age group in future. PMID- 16249156 TI - The effect of remittances on emigration intentions in Egypt, Morocco, and Turkey. AB - Are remittances determined by altruism or enlightened self-interest, and do they trigger additional migration? In this paper these two questions are examined empirically in relation to data from Egypt, Turkey, and Morocco for households with family members living abroad. It is shown, firstly, that one cannot argue exclusively either for altruism or self-interest as motives, since for each country the data tell a different story and both motives can be identified as driving forces behind remittance behaviour. The general conclusion of this study is that the family ties and the net earnings potential of migrants have stronger effects on the flow of remittances than the net earnings potential of the households in the country of origin. Secondly, because the receipt of remittances has a positive effect on the emigration intentions of household members still living in the country of origin, the receipt of remittances may contribute to new flows of migration, particularly in the case of Morocco. PMID- 16249159 TI - Huge differences in cellular radiosensitivity due to only very small variations in double-strand break repair capacity. AB - The DNA double-strand break (DSB) repair capacity of normal human fibroblasts was compared with that of cell lines with different genetic alterations. These cell lines are affected either in non-homologous end-joining (180BR), homology directed repair (C2352, C2395), base excision repair (CS1TAN, 46BR) or signalling (AT3, AT2BE, LFS2675, LFS2800, 95P558). Cellular radiosensitivity was determined by colony formation assay, DSB by constant-field gel electrophoresis and apoptosis was detected by caspase3 activity. For the mutated cell lines, the survival fraction at 2 Gy (SF2) varied between 0.013 and 0.49 in contrast to a variation of only 0.15-0.53 for normal fibroblasts. There was no variation in the number of initial DSB and only a small variation in the number of DSB remaining 24 h after irradiation. At 100 Gy, the latter number varied between 2 and 5 Gy equivalents for normal fibroblasts and only between 3 and 7 Gy-equivalents for the mutated cell lines, corresponding to repair capacities of 95-98 and 93-97%, respectively. There were, however, two outliers (LFS2800, 180BR) where the number of remaining DSB was much higher with 22 and 30 Gy-equivalents, respectively. This elevated number resulted from a delayed repair and apoptotic cells. For all but these two cell lines, the relationship between the number of DSB remaining 24 h after irradiation and SF2 could be described by an identical correlation (r2 = 0.86, p < 0.0001). This result indicates that the relationship between DSB repair capacity and cellular radiosensitivity appears to be the same for normal and mutated cell lines, and that in both cases huge differences in cellular radiosensitivity result from only a very small variation in DSB repair capacity. PMID- 16249160 TI - Retrospective analysis of double-strand break rejoining data collected using warm lysis PFGE protocols. AB - Sample preparation procedures for the pulsed-field gel electrophoresis (PFGE) assay usually involve a lysis step at temperatures as high as 50 degrees C. During this warm-lysis procedure, multiply damaged sites containing heat-labile sites (HLS) can be converted into double-strand breaks (DSB). Once formed, these DSB cannot be distinguished from the DSB formed directly by ionizing radiation. This paper develops a method to correct DSB estimates for the effects of HLS in warm-lysis protocols. A first-order repair model is used to predict the number of HLS available for conversion into DSB as a function of the time available for repair before initiating warm-lysis. A mathematical expression is derived to separate prompt DSB from those formed through the artefactual conversion of HLS into DSB. The proposed formalism only requires the specification of two adjustable parameters, both of which can be estimated from measured data. Estimates of prompt DSB yields obtained by correcting warm-lysis data are in good agreement with estimates obtained using cold-lysis protocols, which do not include the effect of HLS. The retrospective analyses of two published datasets suggest that corrections for HLS have a substantial impact on DSB yields within the first 20-30 min after irradiation. Bi-exponential fits to the DSB data for Chinese hamster ovary cells suggest that corrections for HLS reduce the half-time for fast DSB rejoining by about 15%, whereas the half-time for the slow DSB rejoining only decreases by 4%. The total DSB yield and the fraction of fast rejoining DSB decrease by 24 and 38%, respectively, when the correction is applied. The proposed formalism can be used to characterize trends and uncertainties in DSB rejoining kinetics associated with the artefactual conversion of HLS into DSB. The retrospective application of the methodology to warm-lysis data enhances their relevance and usefulness for studies of DSB rejoining kinetics. PMID- 16249161 TI - Radiation-induced apoptosis in peritoneal resident macrophages of C3H mice: selective involvement of superoxide anion, but not other reactive oxygen species. AB - Remarkably, apoptosis was induced by gamma-ray-irradiation in peritoneal resident macrophages (PRM) of C3H mice, but not other strains of mice. The mechanism of this strain-specific apoptosis induction was studied. Apoptosis in PRM was detected microscopically. Various radical scavengers were examined to identify the critical radicals involved in apoptosis induction. Intracellular peroxide levels were measured with a redox-sensitive dye, 2',7'-dichlorofluorescin diacetate (DCFH). Superoxide dismutase or catalase was introduced into the cells using commercially available Hemagglutinating Virus of Japan (HVJ) envelope vector kit. The enzyme activity of superoxide dismutase was also measured. Radiation-induced apoptosis in C3H mouse PRM was significantly suppressed by treatment with a pharmacological scavenger of superoxide anion, Tiron, but not with other radical scavengers. Intracellular peroxide levels were not elevated by irradiation at doses high enough to induce apoptosis maximally. Radiation-induced apoptosis in C3H mouse PRM was markedly suppressed by superoxide dismutase introduced into the cells using the HVJ envelope vector, but not catalase. The enzyme activity of superoxide dismutase in C3H mouse PRM was comparable with that in B6 mouse PRM. It was concluded that superoxide played the major role in radiation-induced apoptosis in the C3H mouse PRM and that cellular responses downstream or unrelated to superoxide might be responsible for the strain difference in radiation-induced apoptosis of mouse PRM. PMID- 16249162 TI - Absorption of uranium through the entire gastrointestinal tract of the rat. AB - The aim was to determine the gastrointestinal segments preferentially implicated in the absorption of uranium. The apparent permeability to uranium (233U) was measured ex vivo in Ussing chambers to assess uranium passage in the various parts of the small and large intestines. The transepithelial electrical parameters (potential difference, short-circuit current, transepithelial resistance and tissue conductance) were also recorded for each segment. Determination of in vivo uranium absorption after in-situ deposition of 233U in digestive segments (buccal cavity, ileum and proximal colon) and measurements of uranium in peripheral blood were then made to validate the ex vivo results. In addition, autoradiography was performed to localize the presence of uranium in the digestive segments. The in vivo experiments indicated that uranium absorption from the digestive tract was restricted to the small intestine (with no absorption from the buccal cavity, stomach or large intestine). The apparent permeability to uranium measured with ex vivo techniques was similar in the various parts of small intestine. In addition, the experiments demonstrated the existence of a transcellular pathway for uranium in the small intestine. The study indicates that uranium absorption from the gastrointestinal tract takes place exclusively in the small intestine, probably via a transcellular pathway. PMID- 16249163 TI - Evidence-based surgery. PMID- 16249164 TI - Charles R. Drew: surgeon, scientist, and educator. AB - The first African-American Surgeon to achieve extensive training in medical research in the modern era was Dr. Charles Drew (1904-1950) who completed a doctoral degree in medical sciences at Columbia University in 1940. He became the director of the American Red Cross Blood Bank program during World War II and received many accolades for his work with plasma. This historical vignette reviews the details of his life as a scientist and surgical educator while fighting the overpowering racism which black professionals were subjected to during that time. The controversial aspects and facts of his premature death in an automobile accident in the South are explored as well. PMID- 16249165 TI - The effect of G-CSF in an experimental MRSA graft infection in mice. AB - Wound infection after prosthetic material implantation is a troublesome complication with an incidence of 2% to 10%. The effect of granulocyte colony stimulating factor (G-CSF) was studied in an experimental methicillin-resistant Staphylococcus aureus (MRSA) graft infection model. Eighty adult mice were used. Under general anesthesia an abdominal incision of 2 cm in length was performed. A subcutaneous cavity of 2 x 2 cm in size was created. Polypropylene mesh pieces of 2 x 1 cm and MRSA solution of 0.1ml of 10(8) CFU/mL were used. G-CSF was applied systemically or locally in a dosage of 0.02 MU/30 g body weight. There were 8 groups: group I, wound + MRSA; group II, wound + mesh + MRSA; group III, wound + mesh + MRSA + G-CSF (ip, 48 h before operation); group IV, wound + mesh + MRSA + G-CSF (ip, 24 h before operation); group V, wound + mesh + MRSA + G-CSF (locally, into the cavity); group VI, wound + mesh (incubated in G-CSF solution for 4 h) + MRSA; group VII, wound + mesh + MRSA + G-CSF, ip, 24 h from operation; and group VIII (positive control group), wound + mesh + MRSA + Teicoplanin (0.03 mg/30 g body weight, ip, 1/2 h before operation). Three days after, animals were killed and incisions were examined for possible infection or abscess formation and wound failure. Meshes were removed; after vortexing and dilution, samples were incubated with 5% agar media. Results of bacterial incubation were evaluated 24 h and 48 h later. There were symptoms of wound infection and abscess formation in all groups except group VIII. In group VIII, MRSA was isolated in 7 events with a colony count below 10(3). Bacterial counts were above 10(6) (10(6)-10(8)) in all other groups. Thus, it was observed that wound infection could be created with this model, but G-CSF could not prevent the development of wound infection, whether it was administered systemically or locally. Teicoplanin decreased the number of colony-forming units of MRSA, and prevents wound infection in this MRSA wound infection model. PMID- 16249167 TI - Relationship between upper airway obstruction and gastroesophageal reflux in a dog model. AB - The association between gastroesophageal reflux (GER) and upper airway obstruction in children is recognized but not well understood. Our objective was to determine if the creation of a model of upper airway obstruction in dogs would cause GER and to determine if the GER is related to intrathoracic pressure changes. Five dogs underwent evaluation with esophageal manometry and pH probe at baseline and 1 week after creation of an upper airway obstruction. Airway obstruction was created by placement of a fenestrated cuffed tracheostomy tube, which was then capped and the cuff was inflated, requiring the animals to breathe via the fenestrations. The negative inspiratory pressure (Pes) (+/- SD) increased from 11.8 +/- 4.8 cm H(2)O at baseline to 17.6 +/- 4.9 cm H(2)O 1 week after creation of an airway obstruction (p = .029). None of the dogs had GER at baseline with a reflux index (RI) value of 0.0; however, 1 week after creation of airway obstruction, three out of five dogs had GER, with a mean RI value of 21.2 +/- 21.2. There was a significant (p = .023) correlation (r = .928) of the changes in Pes and RI values following airway obstruction. Upper airway obstruction (UAO) does cause GER in this canine model. Severity of GER is significantly correlated with Pes changes. PMID- 16249166 TI - Analysis of bacteremia occurring in the presence of obstruction of the left colon in rats submitted to transoperative antegrade mechanical lavage. AB - With the objective of determining the association between bacteremia and transoperative antegrade mechanical lavage of the colon in an experimental model of obstruction of the left colon in rats, 40 male Wistar rats aged between 90 and 120 days were divided randomly into four groups: A, with intestinal obstruction and with mechanical lavage of the colon; B, with intestinal obstruction and without mechanical lavage of the colon; C, without intestinal obstruction and with mechanical lavage of the colon; and D, without intestinal obstruction and without mechanical lavage of the colon. Analysis of the results showed that there was no bacteremia in the animals in the sham group. On the other hand, bacterial growth in blood cultures was found in three animals (30%) in group C and in four animals (40%) in group B. Positive blood culturing was presented by eight animals (80%) of the rats in group A, and variance analysis on this finding was statistically significant (p = .0029). It can be concluded that, in this experimental model, intestinal obstruction causes a fourfold increase in the risk of bacteremia, while lavage causes an almost threefold increase in the chance of bacterial dissemination into the blood stream. This explains why there was greater incidence of bacteremia in the animals with obstruction and with lavage. PMID- 16249168 TI - The effect of platelet activating factor antagonist BN 52021 on bacterial translocation and ICAM-I expression in experimental obstructive jaundice. AB - Expression of intracellular adhesion molecule-1 (ICAM-1) in an obstructive jaundice model and the potential protective role of platelet activating factor antagonist over small intestine and liver together with its effects on bacterial translocation are examined in this study. Forty-eight male Wistar albino rats were assigned into four equal groups of 12. In groups I and II, animals were sham operated. In groups III and IV, common bile duct ligation and division were performed. In group I and group III, 0.5 ml/day normal saline was applied intraperitoneally daily from day 2 to 6 of the study; in group II and group IV, 1 mg/kg/day BN 52021 was applied intraperitoneally daily from day 2 to 6 of the study. All animals were sacrificed on postoperative day 7. ICAM-1 expression (CD54 positivity) was analyzed in the liver and ileum tissue by immunohistochemical method. Samples from blood, liver mesenteric lymph nodes, and spleen were cultured under aerobic conditions. It is revealed that ICAM-1 expression was statistically higher in group III, with highest bacterial translocation and liver and spleen injury when compared to other groups. Serum alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP), gamma-glutamyltranspeptidase (GGT), bilirubin, tumor necrosis factor alpha (TNFalpha), and interleukin 1beta(IL-1beta) values were at the highest level in group III, and there was a statistical decrease in group IV compared to group III. The administration of BN52021 in experimental obstructive jaundice is a useful way to reduce liver and intestinal mucosal villi damage by inhibiting bacterial translocation and systemic inflammatory response. PMID- 16249169 TI - CT-Directed microcoil localization of small peripheral lung nodules: a feasibility study in pigs. AB - Platinum microcoils were placed in porcine lungs to determine the feasibility for use as a lung nodule marker. Using computed tomography (CT) guidance, the microcoils were successfully deployed in 17 out of 19 attempts. Coil deployment depth ranged from 7 mm to 34 mm below the pleural surface. Moderate pneumothorax was detected after 3 of 19 microcoil insertions. No hemothorax or significant pulmonary hemorrhage was noted. Fluoroscopic guided thoracoscopic resection was successful in 10 of 12 attempts. Platinum microcoils can be safely and easily deployed into the lung parenchyma with minimal complication risk, and can be used to guide subsequent thoracoscopic wedge resection. PMID- 16249170 TI - The influence of inguinal mesh repair on the spermatic cord: a pilot study in the rabbit. AB - The permanent implantation of a polypropylene mesh during inguinal hernia repair causes chronic inflammatory changes in the surrounding tissue. We investigated the effect of this foreign body reaction on the structures of the spermatic cord in the rabbit. Eight Chinchilla rabbits underwent unilateral inguinal hernia repair by the Lichtenstein technique using Marlex (n = 4) or Ultrapro (n = 4) mesh. The contralateral side was operated upon using the Shouldice repair. Three animals served as controls. Three months after operation we analyzed testicular size, testicular temperature, and arterial perfusion by excitation light of a 780 nm laser after injection of 0.5 mg/kg indocyanin green. Histological evaluation included spermatogenesis (Johnsen score) and foreign-body reaction. Testicular volume increased about 10% after each operation. The decrease of arterial perfusion and testicular temperature was more significant after mesh repair than following Shouldice operation. After mesh implantation we found fewer seminiferous tubules classified as Johnsen 10 (Marlex: 51.3%, Ultrapro: 45.0%) than after Shouldice repair (63.8%) or in the controls (65.8%). The spermatic cord showed a typical foreign-body reaction at the interface between mesh and surrounding tissue, which was not detectable after Shouldice repair. Preserved cremasteric muscle fibers protected the structures of the spermatic cord. The inflammatory foreign-body reaction of the surrounding tissue induced by the inguinal prosthetic mesh includes the structures of the spermatic cord. This may have an influence also on spermatogenesis. Therefore, we recommend strict indications for implantation of a prosthetic mesh during inguinal hernia repair. PMID- 16249172 TI - Parts per million mass accuracy on an Orbitrap mass spectrometer via lock mass injection into a C-trap. AB - Mass accuracy is a key parameter of mass spectrometric performance. TOF instruments can reach low parts per million, and FT-ICR instruments are capable of even greater accuracy provided ion numbers are well controlled. Here we demonstrate sub-ppm mass accuracy on a linear ion trap coupled via a radio frequency-only storage trap (C-trap) to the orbitrap mass spectrometer (LTQ Orbitrap). Prior to acquisition of a spectrum, a background ion originating from ambient air is first transferred to the C-trap. Ions forming the MS or MS(n) spectrum are then added to this species, and all ions are injected into the orbitrap for analysis. Real time recalibration on the "lock mass" by corrections of mass shift removes mass error associated with calibration of the mass scale. The remaining mass error is mainly due to imperfect peaks caused by weak signals and is addressed by averaging the mass measurement over the LC peak, weighted by signal intensity. For peptide database searches in proteomics, we introduce a variable mass tolerance and achieve average absolute mass deviations of 0.48 ppm (standard deviation 0.38 ppm) and maximal deviations of less than 2 ppm. For tandem mass spectra we demonstrate similarly high mass accuracy and discuss its impact on database searching. High and routine mass accuracy in a compact instrument will dramatically improve certainty of peptide and small molecule identification. PMID- 16249173 TI - Proteomic analysis of brain plasma membranes isolated by affinity two-phase partitioning. AB - A comprehensive analysis of plasma membrane proteins is essential to in-depth understanding of brain development, function, and diseases. Proteomics offers the potential to perform such a comprehensive analysis, yet it requires efficient protocols for the purification of the plasma membrane compartment. Here, we present a novel and efficient protocol for the separation and enrichment of brain plasma membrane proteins. It lasts only 4 h and is easy to perform. It highly enriches plasma membrane proteins and can be applied to small amounts of brain tissue, such as the cerebellum of a single rat, which was used in the present study. The protocol is based on affinity partitioning of microsomes in an aqueous two-phase system. Marker enzyme assays demonstrated a more than 12-fold enrichment of plasma membranes and a strong reduction of other compartments, such as mitochondria and the endoplasmic reticulum. 506 different proteins were identified when the enriched proteins underwent LC-MS/MS analysis subsequent to protein separation by SDS-PAGE. Using gene ontology, 146 proteins were assigned to a subcellular compartment. Ninety-three of those (64%) were membrane proteins, and 49 (34%) were plasma membrane proteins. A combined literature and database search for all 506 identified proteins revealed subcellular information on 472 proteins, of which 197 (42%) were plasma membrane proteins. These comprised numerous transporters, channels, and neurotransmitter receptors, e.g. the inward rectifying potassium channel Kir7.1 and the cerebellum-specific gamma aminobutyric acid receptor GABRA6. Surface proteins involved in cell-cell contact and disease-related proteins were also identified. Six of the 146 assigned proteins were derived from mitochondrial membranes and 5 from membranes of the endoplasmic reticulum. Taken together, our protocol represents a simple, rapid, and reproducible tool for the proteomic characterization of brain plasma membranes. Because it conserves membrane structure and protein interactions, it is also suitable to enrich multimeric protein complexes from the plasma membrane for subsequent analysis. PMID- 16249174 TI - Sequences required for the activity of PTOX (IMMUTANS), a plastid terminal oxidase: in vitro and in planta mutagenesis of iron-binding sites and a conserved sequence that corresponds to Exon 8. AB - The thylakoid membranes of most photosynthetic organisms contain a terminal oxidase (PTOX, the product of the Arabidopsis IMMUTANS gene) that functions in the oxidation of the plastoquinone pool. PTOX and AOX are diiron carboxylate proteins, and based on crystal structures of other members of this protein class, a structural model of PTOX has been proposed in which the ligation sphere of the diiron center is composed of six conserved histidine and glutamate residues. We tested the functional significance of these residues by site-directed mutagenesis of PTOX in vitro and in planta, taking advantage null immutans alleles for the latter studies. These experiments showed that the six iron-binding sites do not tolerate change, even conservative ones. We also examined the significance of a conserved sequence in (or near) the PTOX active site that corresponds precisely to Exon 8 of the IM gene. In vitro and in planta mutagenesis revealed that conserved amino acids within this domain can be altered but that deletion of all or part of the domain abolishes activity. Because protein accumulates normally in the deletion mutants, the data suggest that the conformation of the Exon 8 sequence is important for PTOX activity. An allele of immutans (designated 3639) was identified that lacks the Exon 8 sequence; it does not accumulate PTOX protein. Chloroplast import assays revealed that mutant enzymes lacking Exon 8 have enhanced turnover. We conclude that the Exon 8 domain is required not only for PTOX activity but also for its stability. PMID- 16249175 TI - The rac activator Tiam1 is a Wnt-responsive gene that modifies intestinal tumor development. AB - Mutations in the canonical Wnt signaling pathway leading to its activation are known to cause the majority of intestinal tumors. However, few genes targeted by this pathway have been demonstrated to affect tumor development in vivo. Here we show that Tiam1, a selective Rac GTPase activator, is a Wnt-responsive gene expressed in the base of intestinal crypts and up-regulated in mouse intestinal tumors and human colon adenomas. Moreover, by comparing tumor development in APC mutant Min (multiple intestinal neoplasia) mice expressing or lacking Tiam1, we found that Tiam1 deficiency significantly reduces the formation and growth of polyps in vivo. However, invasion of malignant intestinal tumors is enhanced by a lack of Tiam1. In line with this, knock-down of Tiam1 reduced the growth potential of human colorectal cancer cells and their ability to form E-cadherin based adhesions, a prerequisite for local invasion of tumor cells. Our data indicate a novel cross-talk between Tiam1-Rac and canonical Wnt-signaling pathways that influences intestinal tumor formation and progression. PMID- 16249176 TI - The plasma membrane Ca2+-ATPase isoform 4 is localized in lipid rafts of cerebellum synaptic plasma membranes. AB - Here we describe the association of the synaptosomal plasma membrane Ca2+-ATPase (PMCA) from pig cerebellum with cholesterol/sphingomyelin-rich membrane domains (rafts). The PMCA4 was localized exclusively in rafts prepared by flotation in Nycodenz density gradients of ice-cold Brij 96 extracts. This was corroborated by its colocalization with the raft markers cholesterol, ganglioside GM1, and PrP(C). The remaining PMCA isoforms were found in the detergent-soluble fractions, with the majority of the membrane proteins. Activity assays confirmed the bimodal distribution of the PMCA isoforms in the density gradient, with a lower activity for PMCA4 and greater stimulation by calmodulin than for the other isoforms. By providing an ordered membrane microenvironment, lipid rafts may contribute to the interaction of PMCA4 with proteins involved in Ca2+ signaling at discrete functional positions on the synaptic nerve terminals. PMID- 16249177 TI - Lipid and peptide control of phosphatidylinositol 4-kinase IIalpha activity on Golgi-endosomal Rafts. AB - The most abundant and widely expressed mammalian phosphoinositide kinase activity is contributed by phosphatidylinositol 4-kinase IIalpha (PI4KIIalpha). In this study we demonstrate that PI4KIIalpha is a novel GTP-independent target of the wasp venom tetradecapeptide mastoparan and that different mechanisms of activation occur in different subcellular membranes. Following cell membrane fractionation mastoparan specifically stimulated a high activity Golgi/endosomal pool of PI4KIIalpha independently of exogenous guanine nucleotides. Conversely, GTPgammaS stimulated a low activity pool of PI4KIIalpha in a separable dense membrane fraction and this response was further enhanced by mastoparan. Overexpression of PI4KIIalpha increased the basal phosphatidylinositol 4-kinase activity of each membrane pool, as well as the mastoparan-dependent activities, thereby demonstrating that mastoparan specifically activates this isozyme. Both mastoparan and M7, at concentrations known to invoke secretion, stimulated PI4KIIalpha with similar efficacies, resulting in an increase in the apparent V(max) and decrease in K(m) for exogenously added PI. Mastoparan also stimulated PI4KIIalpha immunoprecipitated from the raft fraction, indicating that PI4KIIalpha is a direct target of mastoparan. Finally we reveal a striking dependence of both basal and mastoparan-stimulated PI4KIIalpha activity on endogenous cholesterol concentration and therefore conclude that changes in membrane environment can regulate PI4KIIalpha activity. PMID- 16249179 TI - Inhibition of acetyl-CoA carboxylase isoforms by pyridoxal phosphate. AB - Mammalian isoforms of acetyl-CoA carboxylase (ACC-1 and ACC-2) play important roles in synthesis, elongation, and oxidation of long-chain fatty acids, and the possible significance of ACC in the development of obesity has led to interest in the development of inhibitors. Here, we demonstrate that pyridoxal phosphate (PLP) is a linear and reversible inhibitor of ACC-1 and ACC-2. ACC from rat liver and white adipose tissue (largely ACC-1) exhibited an IC50 of approximately 200 microm, whereas ACC-2 from heart or skeletal muscle exhibited an IC50 exceeding 500 microm. ACC from rat liver was equally sensitive to PLP following extensive purification by avidin affinity chromatography. When added before citrate, PLP inhibited ACC with a Ki of approximately 100 microm, reducing maximal activity >90% and increasing the Ka for citrate approximately 5-fold but having little effect on substrate Km values. Pre-treatment with citrate increased the apparent Ki for ACC inhibition by PLP by approximately 4-fold. Inhibition of ACC was reversed by removal of PLP, either by washing or by reaction with hydroxylamine or amino-oxyacetate. ACC was irreversibly inhibited and radiolabeled, to a stoichiometry of approximately 0.4 mol[H]/mol subunit, in the presence of PLP plus [3H]borohydride. Studies with structurally related compounds demonstrated that the reactive aldehyde and negatively charged substituents of PLP contribute importantly to ACC inhibition. The studies reported here suggest a rationale to develop ACC inhibitors that are not structurally related to the substrates or products of the reaction and an approach to probe the citrate-binding site of the enzyme. PMID- 16249178 TI - Cooperative actions of Tra2alpha with 9G8 and SRp30c in the RNA splicing of the gonadotropin-releasing hormone gene transcript. AB - In earlier studies, we demonstrated that excision of the first intron (intron A) from the gonadotropin-releasing hormone (GnRH) transcript is highly cell type- and developmental stage-specific. The removal of GnRH intron A requires exonic splicing enhancers on exons 3 and 4 (ESE3 and ESE4, respectively). Tra2alpha,a serine/arginine-rich (SR)-like protein, specifically binds to ESE4, although it requires additional nuclear co-factors for efficient removal of this intron. In the present study, we demonstrate the cooperative action of multiple SR proteins in the regulation of GnRH pre-mRNA splicing. SRp30c specifically binds to both ESE3 and ESE4, whereas 9G8 binds to an element in exon 3 and strongly enhances the excision of GnRH intron A in the presence of minimal amount of other nuclear components. Interestingly, Tra2alpha can interact with either 9G8 or SRp30c, whereas no interaction between 9G8 and SRp30c is observed. Tra2alpha has an additive effect on the RNA binding of these proteins. Overexpression or knock down of these three proteins in cultured cells further suggests their essential role in intron A excision activities, and their presence in GnRH neurons of the mouse preoptic area further strengthens this possibility. Together, these results indicate that interaction of Tra2alpha with 9G8 and SRp30c appears to be crucial for ESE-dependent GnRH pre-mRNA splicing, allowing efficient generation of mature mRNA in GnRH-producing cells. PMID- 16249180 TI - The crystal structure of the Bacillus anthracis spore surface protein BclA shows remarkable similarity to mammalian proteins. AB - The lethal disease anthrax is propagated by spores of Bacillus anthracis, which can penetrate into the mammalian host by inhalation, causing a rapid progression of the disease and a mostly fatal outcome. We have solved the three-dimensional structure of the major surface protein BclA on B. anthracis spores. Surprisingly, the structure resembles C1q, the first component of complement, despite there being no sequence homology. Although most assays for C1q-like activity, including binding to C1q receptors, suggest that BclA does not mimic C1q, we show that BclA, as well as C1q, interacts with components of the lung alveolar surfactant layer. Thus, to better recognize and invade its hosts, this pathogenic soil bacterium may have evolved a surface protein whose structure is strikingly close to a mammalian protein. PMID- 16249181 TI - Significance of sterol structural specificity. Desmosterol cannot replace cholesterol in lipid rafts. AB - Desmosterol is an immediate precursor of cholesterol in the Bloch pathway of sterol synthesis and an abundant membrane lipid in specific cell types. The significance of the difference between the two sterols, an additional double bond at position C24 in the tail of desmosterol, is not known. Here, we provide evidence that the biophysical and functional characteristics of the two sterols differ and that this is because the double bond at C24 significantly weakens the sterol ordering potential. In model membranes, desmosterol was significantly weaker than cholesterol in promoting the formation or stability of ordered domains, and in mammalian cell membranes, desmosterol associated less avidly than cholesterol with detergent-resistant membranes. Atomic scale molecular dynamics simulations showed that the double bond gives rise to additional stress in the tail, creating a rigid structure between C24 and C27 and favoring tilting of desmosterol distinct from cholesterol. Functional effects of desmosterol in cell membranes were assessed upon acutely exchanging approximately 70% of cholesterol to desmosterol. This led to impaired raft-dependent signaling via the insulin receptor, whereas non-raft-dependent protein secretion was not affected. We suggest that the choice of cholesterol synthesis route may provide a physiological mechanism to modulate raft-dependent functions in cells. PMID- 16249182 TI - Biosensors to measure inositol 1,4,5-trisphosphate concentration in living cells with spatiotemporal resolution. AB - Phosphoinositides participate in many signaling cascades via phospholipase C stimulation, which hydrolyzes phosphatidylinositol 4,5-bisphosphate, producing second messengers diacylglycerol and inositol 1,4,5-trisphosphate (InsP3). Destructive chemical approaches required to measure [InsP3] limit spatiotemporal understanding of subcellular InsP3 signaling. We constructed novel fluorescence resonance energy transfer-based InsP3 biosensors called FIRE (fluorescent InsP3 responsive element) by fusing plasmids encoding the InsP3-binding domain of InsP3 receptors (types 1-3) between cyan fluorescent protein and yellow fluorescent protein sequences. FIRE was expressed and characterized in COS-1 cells, cultured neonatal cardiac myocytes, and incorporated into an adenoviral vector for expression in adult cardiac ventricular myocytes. FIRE-1 exhibits an approximately 11% increase in the fluorescence ratio (F530/F480) at saturating [InsP3] (apparent K(d) = 31.3 +/- 6.7 nm InsP3). In COS-1 cells, neonatal rat cardiac myocytes and adult cat ventricular myocytes FIRE-1 exhibited comparable dynamic range and a 10% increase in donor (cyan fluorescent protein) fluorescence upon bleach of yellow fluorescent protein, indicative of fluorescence resonance energy transfer. In FIRE-1 expressing ventricular myocytes endothelin-1, phenylephrine, and angiotensin II all produced rapid and spatially resolved increases in [InsP3] using confocal microscopy (with free [InsP3] rising to approximately 30 nm). Local entry of intracellular InsP3 via membrane rupture by a patch pipette (containing InsP3)in myocytes expressing FIRE-1 allowed detailed spatiotemporal monitoring of intracellular InsP3 diffusion. Both endothelin-1 induced and direct InsP3 application (via pipette rupture) revealed that InsP3 diffusion into the nucleus occurs with a delay and blunted rise of [InsP3] versus cytosolic [InsP3]. These new biosensors allow studying InsP3 dynamics at high temporal and spatial resolution that will be powerful in under-standing InsP3 signaling in intact cells. PMID- 16249183 TI - Acute physical stress elevates mouse period1 mRNA expression in mouse peripheral tissues via a glucocorticoid-responsive element. AB - In mammals, the circadian and stress systems (both centers of which are located in the hypothalamus) are involved in adaptation to predictable and unpredictable environmental stimuli, respectively. Although the interaction and relationship between these two systems are intriguing and have been studied in different ways since the "pre-clock gene" era, the molecular interaction between them remains largely unknown. Here, we show by systematic molecular biological analysis that acute physical stress elevated only Period1 (Per1) mRNA expression in mouse peripheral organs. Although behavioral rhythms in vivo and peripheral molecular clocks are rather stable against acute restraint stress, the results of a series of promoter analyses, including chromatin immunoprecipitation assays, indicate that a glucocorticoid-responsive element in the Per1 promoter is indispensable for induction of this mRNA both in vitro and in vivo. These results suggest that Per1 can be a potential stress marker and that a third pathway of Per1 transcriptional control may exist in addition to the clock-regulated CLOCK BMAL1/E-box and light-responsive cAMP-responsive element-binding protein/cAMP responsive element pathways. PMID- 16249184 TI - Liver X receptor alpha interferes with SREBP1c-mediated Abcd2 expression. Novel cross-talk in gene regulation. AB - The peroxisomal ATP binding cassette (ABC) transporter adrenoleukodystrophy related protein, encoded by ABCD2, displays functional redundancy with the X linked adrenoleukodystrophy-associated protein, making ABCD2 up-regulation of therapeutic value. Cholesterol lowering activates human ABCD2 in cultured cells. To investigate in vivo regulation by sterols, we first characterized a sterol regulatory element (SRE) in the murine Abcd2 promoter that is directly bound by SRE-binding proteins (SREBPs). Intriguingly, this element overlaps with a direct repeat 4, which serves as binding site for liver X receptor (LXR)/retinoid X receptor heterodimers, suggesting novel cross-talk between SREBP and LXR/retinoid X receptor in gene regulation. Using fasting-refeeding and cholesterol loading, SREBP accessibility to the SRE/direct repeat 4 was tested. Results suggest that adipose Abcd2 is induced by SREBP1c, whereas hepatic Abcd2 expression is down regulated by concurrent activation of LXRalpha and SREBP1c. In cell culture, SREBP1c-mediated Abcd2 induction is counteracted by ligand-activated LXRalpha. Finally, hepatic Abcd2 expression in LXRalpha,beta-deficient mice is inducible to levels vastly exceeding wild type. Together, we identify LXRalpha as negative modulator of Abcd2, acting through a novel regulatory mechanism involving overlapping SREBP and LXRalpha binding sites. PMID- 16249186 TI - Structure of the human Papillomavirus E7 oncoprotein and its mechanism for inactivation of the retinoblastoma tumor suppressor. AB - The E7 oncoprotein from human Papillomavirus (HPV) mediates cell transformation in part by binding to the human pRb tumor suppressor protein and E2F transcription factors, resulting in the dissociation of pRb from E2F transcription factors and the premature cell progression into the S-phase of the cell cycle. This activity is mediated by the LXCXE motif and the CR3 zinc binding domain of the E7 protein. In this study we report the x-ray crystal structure of the CR3 region of HPV E7 and a structure-based mutational analysis to investigate its mode of pRb and E2F binding and E2F displacement from pRb. The structure reveals a novel zinc-bound E7-CR3 obligate homodimer that contains two surface patches of sequence conservation. Mutation of residues within these patches reveals that one patch is required for pRb binding, whereas the other is required for E2F binding. We also show that both E7-mediated interactions are required to disrupt pRb.E2F complexes. Based on these studies we present a mechanistic model for how E7 displaces E2F from pRb. Because the CR3 region of HPV E7 has no detectable homology to other human proteins, the structure-function studies presented here provide an avenue for developing small molecule compounds that inhibit HPV-E7-mediated cell transformation. PMID- 16249185 TI - The structure of dimeric ROCK I reveals the mechanism for ligand selectivity. AB - ROCK or Rho-associated kinase, a serine/threonine kinase, is an effector of Rho dependent signaling and is involved in actin-cytoskeleton assembly and cell motility and contraction. The ROCK protein consists of several domains: an N terminal region, a kinase catalytic domain, a coiled-coil domain containing a RhoA binding site, and a pleckstrin homology domain. The C-terminal region of ROCK binds to and inhibits the kinase catalytic domains, and this inhibition is reversed by binding RhoA, a small GTPase. Here we present the structure of the N terminal region and the kinase domain. In our structure, two N-terminal regions interact to form a dimerization domain linking two kinase domains together. This spatial arrangement presents the kinase active sites and regulatory sequences on a common face affording the possibility of both kinases simultaneously interacting with a dimeric inhibitory domain or with a dimeric substrate. The kinase domain adopts a catalytically competent conformation; however, no phosphorylation of active site residues is observed in the structure. We also determined the structures of ROCK bound to four different ATP-competitive small molecule inhibitors (Y-27632, fasudil, hydroxyfasudil, and H-1152P). Each of these compounds binds with reduced affinity to cAMP-dependent kinase (PKA), a highly homologous kinase. Subtle differences exist between the ROCK- and PKA bound conformations of the inhibitors that suggest that interactions with a single amino acid of the active site (Ala215 in ROCK and Thr183 in PKA) determine the relative selectivity of these compounds. Hydroxyfasudil, a metabolite of fasudil, may be selective for ROCK over PKA through a reversed binding orientation. PMID- 16249188 TI - Molecular consequences of PHOX2B missense, frameshift and alanine expansion mutations leading to autonomic dysfunction. AB - Heterozygous mutations of the PHOX2B gene account for a broad variety of disorders of the autonomic nervous system, either isolated or combined, including congenital central hypoventilation syndrome (CCHS), tumours of the sympathetic nervous system and Hirschsprung disease. In CCHS, the prevalent mutation is an expansion of a 20-alanine stretch ranging from +5 to +13 alanines, whereas frameshift and missense mutations are found occasionally. To determine the molecular basis of impaired PHOX2B function, we assayed the transactivation and DNA binding properties of wild-type and mutant PHOX2B proteins. Furthermore, we investigated aggregate formation by proteins with polyalanine tract expansions ranging from +5 to +13 alanines using immunofluorescence of transfected cells and gel filtration of in vitro translated proteins. We found that transactivation of the dopamine beta-hydroxylase promoter by PHOX2B proteins with frameshift and missense mutations was abolished or severely curtailed, as was in vitro DNA binding although the proteins localized to the nucleus. The transactivation potential of proteins with polyalanine tract expansions declined with increasing length of the polyalanine stretch, and DNA binding was affected for an expansion of +9 alanines and above. Cytoplasmic aggregation in transfected cells was only observed for the longest expansions, whereas even the short expansion mutants were prone to form multimers in vitro. Such a tendency to protein misfolding could explain loss of transactivation for alanine expansion mutations. However, additional mechanisms such as toxic gain-of-function may play a role in the pathogenic process. PMID- 16249187 TI - The Smad6-histone deacetylase 3 complex silences the transcriptional activity of the glucocorticoid receptor: potential clinical implications. AB - Glucocorticoids play pivotal roles in the maintenance of homeostasis but, when dysregulated, may also have deleterious effects. Smad6, one of the transforming growth factor beta (TGFbeta) family downstream transcription factors, interacts with the N-terminal domain of the glucocorticoid receptor (GR) through its Mad homology 2 domain and suppresses GR-mediated transcriptional activity in vitro. Adenovirus-mediated Smad6 overexpression inhibits glucocorticoid action in rat liver in vivo, preventing dexamethasone-induced elevation of blood glucose levels and hepatic mRNA expression of phosphoenolpyruvate carboxykinase, a well known rate-limiting enzyme of liver gluconeogenesis. Smad6 suppresses GR-induced transactivation by attracting histone deacetylase 3 to DNA-bound GR and by antagonizing acetylation of histone H3 and H4 induced by p160 histone acetyltransferase. These results indicate that Smad6 regulates glucocorticoid actions as a corepressor of the GR. From our results and known cross-talks between glucocorticoids and TGFbeta family molecules, it appears that the anti glucocorticoid actions of Smad6 may contribute to the neuroprotective, anticatabolic and pro-wound healing properties of the TGFbeta family of proteins. PMID- 16249189 TI - An animal model for Charcot-Marie-Tooth disease type 4B1. AB - Charcot-Marie-Tooth disease (CMT) comprises a family of clinically and genetically very heterogeneous hereditary peripheral neuropathies and is one of the most common inherited neurological disorders. We have generated a mouse model for CMT type 4B1 using embryonic stem cell technology. To this end, we introduced a stop codon into the Mtmr2 locus within exon 9, at the position encoding amino acid 276 of the MTMR2 protein (E276X). Concomitantly, we have deleted the chromosomal region immediately downstream of the stop codon up to within exon 13. The resulting allele closely mimics the mutation found in a Saudi Arabian CMT4B1 patient. Animals homozygous for the mutation showed various degrees of complex myelin infoldings and outfoldings exclusively in peripheral nerves, in agreement with CMT4B1 genetics and pathology. Mainly, paranodal regions of the myelin sheath were affected, with a high degree of quantitative and qualitative variability between individuals. This pathology was progressive with age, and axonal damage was occasionally observed. Distal nerve regions were more affected than proximal parts, in line with the distribution in CMT. However, we found no significant electrophysiological changes, even in aged (16-month-old) mice, suggesting that myelin infoldings and outfoldings per se are not invariably associated with detectable electrophysiological abnormalities. Our animal model provides a basis for future detailed molecular and cellular studies on the underlying disease mechanisms in CMT4B1. Such an analysis will reveal how the disease develops, in particular, the enigmatic myelin infoldings and outfoldings as well as axonal damage, and provide mechanistic insights that may aid in the development of potential therapeutic approaches. PMID- 16249192 TI - Applicability and transferability of interventions in evidence-based public health. AB - The context in which public health programmes operate can play an important role in influencing their implementation and effectiveness. An intervention that has been shown to be effective in one setting may turn out to be ineffective somewhere else, even supposing it can be implemented there. Therefore, systematic reviews of public health interventions should appraise the applicability of the intervention process and the transferability of the intervention effectiveness to other localities. However, applicability and transferability appraisal is seldom reported in systematic reviews of public health and health promotion interventions. This paper aims to introduce an innovative approach to bridging this gap. A list of attributes that may impact on applicability and transferability can be developed, based on knowledge of the proposed intervention. Then the applicability and transferability of the intervention to the local setting can be rated, and given a score, based on knowledge of the local setting. This approach provides a useful tool for evaluating public health interventions and provides a reliable basis for informed decision making in resource-poor settings, where rigorous primary studies are lacking and where very limited resources put a high demand on evidence-based approaches to health promotion. PMID- 16249191 TI - Preterm birth in twins after subfertility treatment: population based cohort study. AB - OBJECTIVES: To assess gestational length and prevalence of preterm birth among medically and naturally conceived twins; to establish the role of zygosity and chorionicity in assessing gestational length in twins born after subfertility treatment. DESIGN: Population based cohort study. SETTING: Collaborative network of 19 maternity facilities in East Flanders, Belgium (East Flanders prospective twin survey). PARTICIPANTS: 4368 twin pairs born between 1976 and 2002, including 2915 spontaneous twin pairs, 710 twin pairs born after ovarian stimulation, and 743 twin pairs born after in vitro fertilisation or intracytoplasmic sperm injection. MAIN OUTCOME MEASURES: Gestational length and prevalence of preterm birth. RESULTS: Compared with naturally conceived twins, twins resulting from subfertility treatment had on average a slightly decreased gestational age at birth (mean difference 4.0 days, 95% confidence interval 2.7 to 5.2), corresponding to an odds ratio of 1.6 (1.4 to 1.8) for preterm birth, albeit confined to mild preterm birth (34-36 weeks). The adjusted odds ratios of preterm birth after subfertility treatment were 1.3 (1.1 to 1.5) when controlled for birth year, maternal age, and parity and 1.6 (1.3 to 1.8) with additional control for fetal sex, caesarean section, zygosity, and chorionicity. Although an increased risk of preterm birth was therefore seen among twins resulting from subfertility treatment, the risk was largely caused by a first birth effect among subfertile couples; conversely, the risk of prematurity was substantially levelled off by the protective effect of dizygotic twinning. CONCLUSIONS: Twins resulting from subfertility treatment have an increased risk of preterm birth, but the risk is limited to mild preterm birth, primarily by virtue of dizygotic twinning. PMID- 16249193 TI - Glomerular crescent-related biomarkers in a murine model of chronic graft versus host disease. AB - BACKGROUND: We examined the alterations in gene expression associated with the development of crescentic glomerulonephritis in murine chronic graft-versus-host disease, a model for human systemic lupus erythematosus. METHODS: The disease was induced in (C57BL/6 x DBA/2) F(1) hybrids by injection of DBA/2 lymphocytes leading to deposition of auto-antibodies in the glomeruli, and a lupus type of nephritis morphologically. After extensive crescent formation at week 9 of disease, cDNA microarray analysis was performed and highly expressed genes were evaluated as molecular markers by real-time reverse transcription-polymerase chain reaction (RT-PCR), in situ hybridization, immunohistochemistry and immunoassay of urine proteins. RESULTS: Six genes, secreted acidic cysteine-rich glycoprotein (Sparc), thymosin beta 10 (Tmsb10), S100 calcium-binding protein A6 (S100a6), annexin A2 (Anxa2), osteopontin (OPN) and lipocalin 2 (Lcn2), were quantified by real-time RT-PCR in laser microdissected glomeruli in a time course manner. Sparc was detected early before the onset of proteinuria and continued to increase throughout the course of the disease. The expression of Tmsb10, S100a6 and Anxa2 coincided with heavy proteinuria. By week 9, OPN and Lcn2 were highly expressed. The expression of proteins encoded by these genes was predominant in the glomerular crescent. The protein levels of Sparc, OPN and Lcn2 in urine were significantly elevated. CONCLUSIONS: These findings implicate these six genes in the development of glomerular crescents. More importantly, detection of Sparc, OPN and Lcn2 in urine may mean that these molecules could serve as important biomarkers for non-invasive diagnosis of glomerular crescents. PMID- 16249194 TI - In vivo and in vitro effects of simvastatin on inflammatory markers in pre dialysis patients. AB - BACKGROUND: The beneficial effects of statins in reducing cardiovascular events have been attributed predominantly to their lipid-lowering effects, recent studies suggest that these effects might be due to their anti-inflammatory properties. We here investigate the in vivo and in vitro effects of simvastatin on cytokine production in pre-dialysis chronic renal failure patients. METHODS: Our clinical study has been designed as a randomized double-blind placebo controlled study. A total of 55 chronic kidney disease (CKD) patients at stages 3 and 4 (mean creatinine clearance 45 ml/min, range 15-60) were randomly assigned to receive simvastatin 40 mg/day or placebo, added to their ongoing treatment, for 6 months. Blood samples were obtained at baseline, and after 3 and 6 months of observation for the determination of lipids, inflammatory markers and renal function. For the in vitro studies, the effect of increasing doses of simvastatin on cytokine production [namely interleukin (IL)-6 and IL-8] in human cultured monocytes from 10 healthy subjects (HS) and 15 CKD patients stimulated by lipopolysaccharide (LPS) was investigated. RESULTS: A significant reduction in total cholesterol from 221+/-44 mg/dl to 184+/-41 mg/dl (3 months) and to 186+/ 39 mg/dl (6 months) (P<0.02) and low-density lipoprotein cholesterol from 139+/ 40 mg/dl to 104+/-29 mg/dl (3 months) and to 100+/-31 mg/dl (6 months) (P<0.001) was observed in the 28 patients treated with simvastatin. In this group, C reactive protein (CRP) levels significantly decreased from 2.6 mg/l [interquartile range (IQR 4.9)] to 2.0 mg/l (IQR 1.9) (P = 0.03) at 6 months (P<0.05). A parallel reduction of IL-6 levels from 5.1 pg/ml (IQR 3.8) to 3.5 pg/ml (IQR 3.1) (P = 0.001) at 6 months was also observed. No significant reduction in inflammatory markers [CRP from 5.1 mg/l (IQR 1.9) to 5.4 mg/l (IQR 1.3) (P = NS) at 6 months] or plasma lipids [LDL-cholesterol from 127+/-32 mg/dl to 131+/-21 mg/dl (6 months)] was observed in the 27 patients of the placebo group. In the in vitro studies, the average value for cell-associated IL-6 and IL 8 was higher in CKD (155+/-95 pg/ml monocytes for IL-6 and 722+/-921 pg/ml monocytes for IL-8) vs HS (137+/-87 pg/ml monocytes and 186+/-125 pg/ml monocytes) (P<0.01) and was not affected by simvastatin alone. LPS resulted in a significant increase in cytokine production (IL-6: 1954+/-321 pg/ml monocytes for CKD and 1451+/-237 pg/ml monocytes for HS; P<0.001); the simultaneous addition of increasing doses of simvastatin to these cultures induced a dose-dependent inhibition of IL-6 and IL-8 production in stimulated peripheral blood mononuclear cells in all groups. CONCLUSIONS: These results indicate that simvastatin in commonly used doses has an in vitro and in vivo anti-inflammatory effect in CKD patients, and may play an important role in counteracting the mechanisms involved on the pathogenesis of cardiovascular disease. PMID- 16249195 TI - Myeloperoxidase serves as a marker of oxidative stress during single haemodialysis session using two different biocompatible dialysis membranes. PMID- 16249196 TI - Home treatment for Fabry disease: practice guidelines based on 3 years experience in The Netherlands. AB - INTRODUCTION: Recently, chronic supplementation with alpha-galactosidase A (alphaGal A) has been approved as a treatment modality for Fabry disease. The aim of the current study was to investigate the feasibility of home therapy for Fabry disease during a follow-up of >3 years and to make a proposal for practice guidelines. METHODS: Based on experience in previous clinical trials, an algorithm for home treatment eligibility was developed. The number of successful and uneventful infusions was recorded, as well as adverse and infusion-associated events. The presence and titre of recombinant human (rh)-alphaGal A antibodies were monitored every 3 months. RESULTS: Thirty of the 36 patients eligible for home treatment received a total of 1418 infusions at home (median 44 infusions, range 1-108), between March 2001 and July 2005. Mean age was 44.7 years (17-71). Seventeen patients receiving home treatment (57%) were male. The majority of patients (27 out of 30, 90%) undergoing home treatment received 0.2 mg/kg agalsidase alpha or beta. Six male patients developed an infusion-associated event, of which three developed these at home. All patients with an infusion associated event were anti-rh-alphaGal A IgG positive at 3 months, but three patients with rh-alphaGal A antibodies did not develop side effects. Antibody titres between these patients did not differ. None of the events was life threatening or necessitated urgent admission. CONCLUSION: Home treatment with rh alphaGal A for Fabry disease with 0.2 mg/kg for males and both 1.0 and 2.0 mg/kg for females is feasible and safe, and reduces both the burden related to chronic intravenous therapy and health care costs. Whether this can also be applied for male patients treated with 1.0 mg/kg has not yet been determined. PMID- 16249197 TI - Acute renal failure following endovascular repair of an infrarenal abdominal aortic aneurysm. PMID- 16249198 TI - A neutralizing VEGF antibody prevents glomerular hypertrophy in a model of obese type 2 diabetes, the Zucker diabetic fatty rat. AB - BACKGROUND: Antagonism of vascular endothelial growth factor (VEGF) has improved the outcome in experimental nephropathies of various origins, including diabetic nephropathy in a type 1 diabetic rat model and a type 2 diabetic mouse model. Neutralizing VEGF antibodies prevented glomerular hypertrophy in these models. We examined the renal effects of VEGF blockade in an obese rat model of type 2 diabetic nephropathy and investigated the mechanism underlying the inhibition of glomerular hypertrophy. METHODS: Twenty female Zucker diabetic fatty (ZDF) rats, fed a high-fat diet and aged 10 weeks, were treated with VEGF antibodies or an irrelevant isotype-matched IgG. Ten heterozygous (fa/+) littermates served as additional non-diabetic, lean controls. Urinary albumin excretion (UAE) and creatinine clearance (CrCl) were assessed at baseline, and at 3 and 5 weeks. Kidney weight and glomerular volume were determined at the end of the study. Glomerular apoptosis was examined with anti-active caspase-3 immunohistochemistry. RESULTS: All obese animals had established diabetes, hyperlipidaemia and normal blood pressure, which were not influenced by VEGF antibody treatment. ZDF control rats had increased UAE, CrCl, kidney weights and glomerular volumes compared with non-diabetic, lean control rats. VEGF antibody treatment prevented the glomerular hypertrophy, but did not affect UAE, CrCl and kidney weight. Glomerular anti-active caspase-3 immunostaining was not different between the groups. CONCLUSIONS: Inhibition of VEGF prevented early glomerular hypertrophy in ZDF rats with established diabetes. Increased apoptosis of glomerular endothelial cells does not appear to underly the inhibition of glomerular growth. PMID- 16249199 TI - Efficacy prospective study of different frequencies of Epo administration by i.v. and s.c. routes in renal replacement therapy patients. AB - BACKGROUND: The problem of pure red cell aplasia (PRCA) prompted nephrologists to revert to a wider intravenous (i.v.) utilization of erythropoeitin (Epo). Once weekly i.v. Epo administration has been suggested to be as effective as the twice/thrice weekly i.v. dose. The aim of the present study was to test whether once weekly i.v. Epo administration is equally as cost-effective as once weekly subcutaneous (s.c.) and 2-3 times weekly i.v. administration. METHODS: We prospectively studied 41 patients (23 males, aged 28-82 years), on renal replacement therapy for 18-286 months, stabilized on twice or thrice weekly s.c. Epo-alpha (basal). The patients were treated for three consecutive 6 month periods with once weekly s.c. (OWSC), once weekly i.v. (OWIV) and twice/thrice weekly i.v. (TWIV) Epo-alpha. The initial dose for each period was equal to the final dose of the previous one; when necessary, the dose was adjusted according to DOQY guidelines. Iron, folic acid and vitamin B(12) supplementations were given throughout all the study periods. At the end of each of the four study periods, the following parameters were evaluated: haemoglobin, haematocrit, hypochromic red blood cells (RBCs), iron, serum ferritin, transferrin, folate, vitamin B(12), C-reactive protein (CRP), Kt/V, parathyroid hormone (PTH) and weekly dose of Epo-alpha. RESULTS: Thirty-three out of 41 enrolled patients completed the study (there were five deaths, two renal transplants and one transfer). No significant changes were observed as regards iron, serum ferritin, transferrin, folate, vitamin B(12), CRP, Kt/V or PTH level. Haemoglobin levels were not different at the end of the basal (11.7+/-1.21), OWSC (11.8+/-0.86) and TWIV (12.1+/-1.04) periods, while significantly lower levels were observed after the OWIV period (11.0+/-0.97, P<0.01). Weekly Epo consumption (Epo U/week/kg body weight/g haemoglobin) was: basal 11.57+/-5.96; OWSC 10.22+/-4.53; OWIV 15.99+/ 7.7*(a); and TWIV 11.89+/-6.3*(a) (*P<0.01 vs basal; (a)P<0.01 vs OWSC). CONCLUSIONS: From our results, the OWIV schedule seems to have less efficacy in the control of anaemia of chronic renal failure patients on dialysis treatment than either OWSC or TWIV schedules. PMID- 16249200 TI - Ultrasonographic measurement of intima-media thickness of radial artery in pre dialysis uraemic patients: comparison with histological examination. AB - BACKGROUND: Increased intima-media thickness (IMT) of the radial artery is associated with early failure of radiocephalic arteriovenous fistula (AVF) in haemodialysis patients. Therefore, non-invasive measurements of radial artery IMT before AVF operations are very important in predicting AVF patency. This study was designed to evaluate the accuracy of high-resolution ultrasonography in measuring radial artery IMT in pre-dialysis uraemic patients. METHODS: This study enrolled 43 pre-dialysis uraemic patients awaiting radiocephalic AVF operations for the first time. In this study, 17 age- and sex-matched uncomplicated hypertensive patients and 15 healthy subjects were included as a control. We measured the internal diameter (ID) and IMT of the radial artery using high resolution ultrasonography on the wrists of uraemic patients as well as the control group before the AVF operation. We obtained specimens of the radial artery during the AVF operation and directly measured the IMT by histological examination. RESULTS: The radial artery IMT of the uraemic patients (0.41 +/- 0.09 mm) was significantly thicker, compared to both those of the hypertensive (0.33 +/- 0.05 mm, P < 0.001) and the healthy patients (0.25 +/- 0.04 mm, P = 0.002). In contrast, the radial artery ID in the uraemic patients (1.85 +/- 0.48 mm) was smaller than both that of the hypertensive patients (2.08 +/- 0.31 mm, P = 0.023) and the healthy persons (2.34 +/- 0.37 mm, P = 0.001). Radial artery IMT had a negative correlation with radial artery ID in a total of 73 subjects (r = 0.290, P = 0.012). The value of the radial arterial IMT measured by sonographic examination correlated significantly with that by histological examination in 43 uraemic patients (r = 0.786, P < 0.001) and it correlated significantly with early AVF failure (r = 0.358, P = 0.027). CONCLUSION: Our data suggest that high resolution ultrasonography is an effective tool in measuring radial artery IMT in uraemic patients before AVF operation. PMID- 16249201 TI - The course of type 1 hepato-renal syndrome post liver transplantation. AB - BACKGROUND: Hepato-renal syndrome (HRS) is a functional form of renal failure that occurs in patients with end-stage liver disease. Previously considered fatal without liver transplantation, treatment with vasoconstrictors and albumin has been demonstrated to improve renal function in patients with type 1 HRS. Liver transplantation is still considered the definitive treatment for HRS. However, the renal recovery rate and those factors that predict recovery post orthotopic liver transplantation have not been determined. METHODS: We reviewed the hospital course of 28 patients who met the International Ascites Club criteria for type I HRS and who underwent orthotopic liver transplant. The patients' demographic and pre- and post-operative laboratory data were recorded; patients were followed for 4 months post-transplantation or until death. RESULTS: The MELD score of the patients was 30+/-6. The mean duration of HRS prior to liver transplantation was 37+/-27 days. HRS resolved in 16 patients (58%). The mean time to resolution of HRS was 21+/-27 days, with a range of 4-110 days. Eight (50%) patients in whom the HRS resolved were undergoing pre-transplantation dialysis. The age of the recipients (49+/-10 vs 56+/-12; P = 0.05), the total bilirubin level on post operative day 7 (6.0+/-4.3 vs 10.1+/-5.9 mg/dl; P = 0.04), alcoholic liver disease and the requirement for post-transplant dialysis were predictors of resolution of HRS by univariate analysis. Only alcoholic liver disease and post transplant dialysis were independent (negative) predictors of resolution of HRS. Seven of the 12 (58%) patients who developed chronic renal insufficiency remained dialysis dependent. The pre-operative serum creatinine was non-significantly higher in the non-resolvers who remained dialysis dependent compared to those who did not require long-term dialysis (3.0+/-1.0 vs 2.3+/-0.4 mg/dl; P = 0.1) Four patients died; in three of these patients the HRS had resolved prior to their death. CONCLUSION: HRS is not always cured by orthotopic liver transplant. Pre transplantation dialysis or a long waiting period should not preclude transplantation in patients with HRS. HRS may not resolve in patients with alcoholic liver disease. We were unable to accurately define that group of patients with HRS who required long-term dialysis and could theoretically benefit from combined liver-kidney transplantation. PMID- 16249202 TI - Therapeutic action of citrate in urolithiasis explained by chemical speciation: increase in pH is the determinant factor. AB - BACKGROUND: The therapeutic action of citrate in the management of calcium oxalate urolithiasis has been attributed to the depletion of free calcium ions by complexation of the latter by citrate itself. However, little attention has been given to the nature of such complexes and the chemical conditions which control their formation because it is very difficult to measure them in solution. We therefore modelled the theoretical formation of these complexes in urine following administration of a citrate-containing preparation, using a powerful speciation program, JESS (Joint Expert Speciation System), which has been widely used to model metal-ligand equilibria in biological systems but which has hitherto not been applied in urolithiasis research. This program has an extensive database of thermodynamic constants and is able to calculate mixed ligand speciation. METHODS: Urine data obtained before and after citrate administration in four groups of subjects (male and female normals and stone formers) were used as input for JESS to calculate the speciation of calcium, citrate and oxalate. The program was also used to examine the effects of varying different urinary components on the nature and concentration of the various species. RESULTS: The speciation predicted the formation of a key calcium-citrate-phosphate species (previously unreported in urolithiasis research), which accounts for a significant percentage of the complexation of the free calcium. Moreover, the formation of this complex was found to be dependent on an increase in urinary pH rather than on an increase in urinary citrate concentration per se. CONCLUSION: The therapeutic action of citrate in the management of calcium oxalate urolithiasis is due to the formation of a pH dependent calcium-citrate-phosphate complex which reduces the concentration of the free calcium ion species, thereby reducing the risk of stone formation. PMID- 16249203 TI - Haemoglobin at time of referral prior to dialysis predicts survival: an association of haemoglobin with long-term outcomes. AB - Haemoglobin (Hgb) levels are known to be associated with numerous adverse outcomes in both chronic kidney disease (CKD) and non-CKD patients. This analysis evaluates the association of baseline haemoglobin levels on survival in CKD patients, who are followed by nephrologists, irrespective of glomerular filtration rate (GFR), prior to initiation of renal replacement therapy (RRT) and erythropoietin hormone replacement therapy. Analysis of data from the provincial database (PROMIS, Patient Registration and Outcome Management Information System) in British Columbia, Canada, was undertaken. Records used for the analysis included all CKD patients at first registration: GFR <60 ml/min/1.73 m(2), not yet on dialysis, starting from May 1998 to October 2002, and who had complete data (defined as age and gender, diabetic status, eGFR and Hgb levels). The primary objective of this study was to determine the association of Hgb and survival controlling for eGFR at first registration value, age, gender and diabetic status. Multivariate Cox proportional hazards analysis with time to death as outcome variable was performed. The cohort included 3028 patients: the mean age was 65 years, 28% were diabetic, and the mean eGFR in the cohort was 21 ml/min/1.73 m(2). The cohort is representative of the BC CKD and dialysis population regarding ethnicity: 64% Caucasian, 32% Asian. Median follow-up was 27 months, 1 year survival was 0.92, 2 year survival was 0.85. Hgb at initial registration is a statistically independent predictor of survival (RR = 0.875 for every 10 g/l, 95% CI: 0.835-0.917, P = 0.0001), after adjusting for age, gender, diabetic status and baseline eGFR. Further analysis, controlling for RRT, demonstrated a similar association between Hgb and survival (RR = 0.853 for every 10 g/l, 95% CI: 0.799-0.910, P = 0.0001), after adjusting for above variables. Substantial variation in Hgb values exists at all GFR levels. These findings underscore the importance of evaluating Hgb at all GFR levels, and the need to study the impact of modification of Hgb at different GFR levels on survival. PMID- 16249204 TI - Epidemiology of renal disease in Romania: a 10 year review of two regional renal biopsy databases. AB - BACKGROUND: Epidemiological data of renal disease are available from large national renal biopsy registries from Central and Western European countries; in contrast, detailed epidemiological data from Eastern European countries are missing. This report is the first review of histological data, over a period of 10 years (1995-2004), covering a population of over 6 million inhabitants and two distinct regions from an East European country - Romania. METHODS: 635 eco-guided kidney biopsies from the Moldova (North-Eastern Romania, 8 counties, 4 754 048 inhabitants) and Banat (Western Romania, 3 counties, 1 454 747 inhabitants) regions were analysed. Data on serum creatinine concentration (sCr), 24 h proteinuria, haematuria, clinical diagnosis, histological diagnosis and complications after renal biopsy were collected. RESULTS: The number of biopsies performed varied between 10.9 p.m.p./year in 1995 and 11.3 p.m.p./year in 2004. The most common clinical syndromes - as indication for performing the renal biopsy - were: nephrotic syndrome (52.3%), followed by nephritic syndrome (21.9%), acute renal failure (ARF) (12.4%), chronic kidney disease (CKD) (10.2%) and asymptomatic urinary abnormalities (AUA) (3.3% of the cases). The major histological groups identified were: primary glomerulonephritis (GN) (66.2%), secondary GN (26.4%), vascular nephropathies (2.3%), and tubulointerstitial nephropathies (TIN) (1.5%) of the cases. Among primary GN's, the most frequent diagnoses were: membranoproliferative GN (MPGN) (29.4%, incidence in 2004 - 9.3 p.m.p./year), mesangioproliferative GN (MesGN) (28.9%, incidence - 10 p.m.p./year), membranous GN (MGN) (11.2%, incidence - 5.3 p.m.p./year), minimal change disease (MCD) (8.5%, incidence - 7.3 p.m.p./year), focal and segmental glomerulosclerosis (FSGS) (11.5%, incidence - 3.3 p.m.p./year) and crescentic GN (CGN) (7.9%, incidence - 3.3 p.m.p./year). The prevalence of membranoproliferative GN significantly decreased from 1995 to 2004. The prevalence of different types of secondary GN was similar to Western and Central European countries, with the particular difference of higher infectious diseases associated GN. CONCLUSION: The present data are an important contribution to the epidemiology of renal diseases in Europe, highlighting not only numerous similarities but also significant epidemiological differences in Western and Central European countries, particularly a higher, albeit declining, incidence and prevalence of membranoproliferative GN. This report represents the basis for the future of Romanian Registry of Renal Biopsies and is intended to serve as a source of information for nephrologists concerned with East European renal pathology. PMID- 16249205 TI - The role of micro-inflammation in the pathogenesis of uraemic pruritus in haemodialysis patients. AB - BACKGROUND: Uraemic pruritus (UP) is still one of the most vexing and disabling symptoms in chronic renal failure. The pathogenesis of UP is obscure and effective therapeutic strategies are elusive. Deduced from partial successful treatment modalities, there is evidence that an alteration of the immune system with a pro-inflammatory pattern along with a deranged T-helper-cell differentiation may be involved in the pathogenesis of UP. We, therefore, investigated whether UP is related to an augmented Th1-differentiation as measured by determination of intracytoplasmatic (i.c.) cytokines and expression of chemokine receptors. Additionally, pro-inflammatory cytokines were determined in serum. METHODS: In a multicentre study, 171 patients on haemodialysis (HD) were screened for UP. Finally, 13 HD patients with and 13 HD patients without UP, as well as 15 healthy controls were enrolled in the study. Peripheral blood mononuclear cells were isolated and the proportion of Th1- and Th2-cells was determined by flow cytometry. The expression of chemokine receptors on CD4 cells (CXCR3 preferentially on Th1 and CCR4 on Th2) and i.c. cytokines (IFNgamma for Th1 and IL4 for Th2) were measured after in vitro stimulation. Serum cytokine levels (IL6 and TNFalpha) and CRP were measured by ELISA. RESULTS: Compared to HD patients without UP, those complaining of UP showed a significantly enhanced proportion of Th1-cells as measured by both techniques. Additionally, serum CRP and IL6 levels were significantly higher in HD patients with UP, compared to HD patients without UP. CONCLUSIONS: These results point to a central role of inflammation in the pathogenesis of UP in HD patients. PMID- 16249206 TI - Gas-forming infection in a renal cyst of a patient with autosomal dominant polycystic kidney disease. PMID- 16249207 TI - Are multiple sclerosis patients risk-takers? AB - Several factors appear to be associated with multiple sclerosis (MS), and each has a postulated immune or environmental explanation, but a common theme is lacking. This article suggests that a unifying premise could be risk-associated behaviour. Evidence is reviewed for associations with smoking, alcohol, recreational drug use, oral contraception, cholesterol intake, risk attitude and behaviour, ultraviolet light and vitamin D exposure, frequency of MS in healthy societies, and viral infection. The evidence associated with smoking, not taking vitamin D supplements and Epstein-Barr viral infection appears good. There may be a pattern of risk-associated behaviour that characterizes patients with MS and brings them into contact with one or more causative agents. Of the possible agents, viral infection seems the most likely. PMID- 16249208 TI - Responses to: "An evaluation of the impact of a US$60 million nutrition programme in Bangladesh". PMID- 16249209 TI - Comment on contributions regarding the impact of the Bangladesh Integrated Nutrition Project. PMID- 16249211 TI - Consumption of sweet foods and breast cancer risk in Italy. AB - BACKGROUND: The relation between the intake of sugar and sweets and the risk of breast cancer has been considered in ecological, prospective and case-control studies, but the results are unclear. We analyzed such a relation in a case control study conducted between 1991 and 1994 in Italy. PATIENTS AND METHODS: Cases were 2569 women with histologically confirmed incident breast cancer and controls were 2588 women admitted to hospital for acute, non-neoplastic, non hormone-related conditions. Information on diet was based on an interviewer administered questionnaire tested for reproducibility and validity. The odds ratios (OR) and 95% confidence intervals (CI) were computed by multiple logistic regression equations. RESULTS: Compared with women with the lowest tertile of intake, women in the highest tertile of intake of desserts (including biscuits, brioches, cakes, puffs and ice-cream) and sugars (including sugar, honey, jam, marmalade and chocolate) had multivariate ORs of 1.19 (95% CI 1.02-1.39) and 1.19 (95% CI 1.02-1.38), respectively. The results were similar in strata of age, body mass index, total energy intake and other covariates. CONCLUSIONS: We found a direct association between breast cancer risk and consumption of sweet foods with high glycemic index and load, which increase insulin and insulin growth factors. PMID- 16249212 TI - Life expectancy as an indicator of outcome in follow-up of population-based cancer registries: the example of childhood leukemia. AB - BACKGROUND: Survival analysis is a standard methodology to assess progress in oncology disease treatment. However, survival analysis commonly only measures survival during the treatment period (and the period immediately afterwards), and does not provide an estimate of life expectancy, which is often of more interest to patients and to health policy makers. In this paper we propose a method to estimate childhood acute lymphoblastic leukemia (ALL) life expectancy through the integration of traditional survival analysis and life expectancy tables. PATIENTS AND METHODS: The study included 305 incident cases registered by the Childhood Cancer Registry of Piedmont in 1979-1991. Vital status on 30 June 2004 was known for 304 cases. Survival analyses were carried out using the Kaplan-Meier method and the Gompertz model, according to the time period of diagnosis and gender. RESULTS: Cumulative survival at 5 years increased from 58.6% (95% CI 48.9-68.3) for cases diagnosed in March 1979-July 1982 to 79.1% (95% CI 70.8-87.5) in March 1987-February 1991 (P = 0.002). Average life expectancy increased from 46.1 years for boys and 42.6 years for girls diagnosed in March 1979-July 1982 to 58.3 and 69.1, respectively, in March 1987-February 1991. CONCLUSIONS: These analyses show an improvement over the time period of diagnosis of life expectancy for children with ALL. PMID- 16249213 TI - Adult height in relation to mortality from 14 cancer sites in men in London (UK): evidence from the original Whitehall study. AB - BACKGROUND: Adult height has been related to organ-specific malignancies in relatively few studies. Findings are discrepant for some sites and several studies are subject to a series of methodological limitations. MATERIALS AND METHODS: We examined the association of adult height with death attributed to 14 cancer sites using data from the original Whitehall cohort. This is a prospective study of 18,403 middle-aged, non-industrial, London-based, male government employees who were examined in the late 1960s and then followed up for mortality for a maximum of 35 years. RESULTS: There were 11,099 deaths during follow-up, 3101 (28%) of which were ascribed to cancer. Cox proportional hazards regression models revealed modest effects for height in relation to site-specific cancers. Following adjustment for covariates that included employment grade (an indicator of socioeconomic position), body mass index and smoking habit, increased height was associated with elevated mortality rates for cancer of combined sites [hazards ratio per 5 cm increase in height (95% confidence interval); P for trend across height categories: 1.05 (1.03, 1.08); P < 0.001], lung [1.13 (1.06, 1.20); P < 0.001], prostate [1.07 (0.99, 1.15); P = 0.08], kidney [1.20 (0.99, 1.46); P = 0.08], skin [1.35 (1.06, 1.70); P = 0.02] and leukaemia [1.11 (0.96, 1.28); P = 0.02]. CONCLUSIONS: Amongst other explanations, the weak positive height-cancer gradients apparent herein may be ascribed to early life exposures that correlate with adult height, such as high caloric intake. PMID- 16249214 TI - Allelic loss analysis of early-stage flat-type colorectal tumors. AB - BACKGROUND: Flat-type colorectal tumors are rare, but are known for their unusual flat morphology and aggressive clinical behavior despite their small size. To identify distinct genetic alterations, loss of heterozygosity (LOH) analysis was performed on microdissected tissues. MATERIALS AND METHODS: DNA was extracted from multiple microdissected foci in 43 cases of early-stage flat-type colorectal tumors and LOH analysis was performed on 2q, 4q, 5q, 12q, 14q, 15q, 17p, 18q, 18p and 22q. RESULTS: LOH patterns were detected in one of two forms: (i) homogeneous LOH throughout the microdissected foci, which indicated the early acquisition of LOH; and (ii) heterogeneous LOH, which were detected in a part of analyzed foci. Homogeneous and heterogeneous LOH were most frequently detected on 17p (92%) followed by 18q (81%), 18p (81%), 5q (61%), 22q (51%), 14q (44%), 15q (41%), 2q (39%), 12q (36%) and 4q (32%). Homogeneous LOH was detected most frequently on 17p (68%) followed by 18p (53%), 18q (53%), 22q (34%) and 12q (27%). The average fractional allelic loss (FAL) for heterogeneous and homogeneous LOH was 0.57 and the average FAL for homogeneous LOH was 0.37. CONCLUSIONS: Early flat-type colorectal tumors frequently shows the early occurrence of multiple LOH including 17p, 18p, 18q and 22q, which is coupled with additional LOH of other loci either simultaneously or in the early clonal progression phase. The extent and sequences of LOH may be the mechanisms responsible for the aggressive clinical behaviors of these tumors. PMID- 16249215 TI - A randomized phase II trial comparing every 3-weeks carboplatin/paclitaxel with every 3-weeks carboplatin and weekly paclitaxel in advanced non-small cell lung cancer. AB - BACKGROUND: The optimal schedule of taxane administration has been an area of active interest in several recent clinical trials. METHODS: To address a pure schedule question, we randomized 161 patients with advanced stage IIIB or IV non small-cell lung cancer (NSCLC) to either paclitaxel 225 mg/m2 every 3 weeks x 4 cycles or 75 mg/m2/week x 12 (cumulative dose on each arm = 900 mg/m2). Both arms received concurrent carboplatin AUC 6 every 3 weeks x 4 cycles. RESULTS: The two arms were well-balanced in terms of known prognostic factors. The overall response rate and survival outcomes were similar on the two arms. There was significantly more grade 3/4 thrombocytopenia and grade 2-4 anemia on the weekly arm but less severe myalgias/arthralgias and alopecia. No difference in the rates of peripheral neuropathy was observed; however, patients on the every 3 weeks arm reported significantly more taxane therapy-related side-effects on the functional assessment of cancer therapy taxane subscale. CONCLUSIONS: This randomized trial exploring schedule-related issues with carboplatin/paclitaxel confirms the versatility of this regimen. PMID- 16249216 TI - Modelling the structure of latexin-carboxypeptidase A complex based on chemical cross-linking and molecular docking. AB - We have determined the three-dimensional structure of the protein complex between latexin and carboxypeptidase A using a combination of chemical cross-linking, mass spectrometry and molecular docking. The locations of three intermolecular cross-links were identified using mass spectrometry and these constraints were used in combination with a speed-optimised docking algorithm allowing us to evaluate more than 3 x 10(11) possible conformations. While cross-links represent only limited structural constraints, the combination of only three experimental cross-links with very basic molecular docking was sufficient to determine the complex structure. The crystal structure of the complex between latexin and carboxypeptidase A4 determined recently allowed us to assess the success of this structure determination approach. Our structure was shown to be within 4 A r.m.s. deviation of Calpha atoms of the crystal structure. The study demonstrates that cross-linking in combination with mass spectrometry can lead to efficient and accurate structural modelling of protein complexes. PMID- 16249217 TI - A mathematical model to estimate global hepatitis B disease burden and vaccination impact. AB - BACKGROUND: Limited data are available regarding global hepatitis B virus (HBV) related morbidity and mortality and potential reduction in disease burden from hepatitis B vaccination. METHODS: A model was developed to calculate the age specific risk of acquiring HBV infection, acute hepatitis B (illness and death), and progression to chronic HBV infection. HBV-related deaths among chronically infected persons were determined from HBV-related cirrhosis and hepatocellular carcinoma (HCC) mortality curves, adjusted for background mortality. The effect of hepatitis B vaccination was calculated from vaccine efficacy and vaccination series coverage, with and without administration of the first dose of vaccine within 24 h of birth (i.e. birth dose) to prevent perinatal HBV infection. RESULTS: For the year 2000, the model estimated 620,000 persons died worldwide from HBV-related causes: 580,000 (94%) from chronic infection-related cirrhosis and HCC and 40,000 (6%) from acute hepatitis B. In the surviving birth cohort for the year 2000, the model estimated that without vaccination, 64.8 million would become HBV-infected and 1.4 million would die from HBV-related disease. Infections acquired during the perinatal period, in early childhood (<5 years old), and > or = 5 years of age accounted for 21, 48, and 31% of deaths, respectively. Routine infant hepatitis B vaccination, with 90% coverage and the first dose administered at birth would prevent 84% of global HBV-related deaths. CONCLUSION: Globally, most HBV-related deaths result from the chronic sequelae of infection acquired in the perinatal and early childhood periods. Inclusion of hepatitis B vaccine into national infant immunization programs could prevent >80% of HBV-related deaths. PMID- 16249218 TI - Tobacco associated mortality in Mumbai (Bombay) India. Results of the Bombay Cohort Study. AB - BACKGROUND: Little is known about the excess mortality from forms of tobacco use other than cigarette smoking that are widely prevalent in India, such as bidi smoking and the various forms of smokeless tobacco use. We report on absolute and relative risks of mortality among various kinds of ever tobacco users vs never users in the city of Mumbai, India. METHODS: Using the Mumbai voters' list as the selection frame, 99 570 individuals aged > or = 35 years were interviewed at their homes during 1992-94. At active follow-up (during 1997-99) after 5.5 years, 97 244 (97.7%) were traced. Among these, 7531 deaths (4119 men, 3412 women) were recorded, of which 89% died within study area. It was possible to abstract cause of death information from the records of the municipal corporation for 5470 deaths. These were coded using ICD 10. RESULTS: The adjusted relative risk was 1.37 (95% CI 1.23-1.53) for (men) cigarette smokers and 1.64 (95% CI 1.47-1.81) for bidi smokers, with a significant dose-response relationship for number of bidis or cigarettes smoked. Women were essentially smokeless tobacco users; the adjusted relative risk was 1.25 (95% CI 1.15-1.35). The risk of deaths from respiratory diseases (RR 2.12, 95% CI 1.57-2.87), tuberculosis (RR 2.30, 95% CI 1.68-3.15), and neoplasms (RR 2.60, 95% CI 1.78-3.80) were significantly high in male smokers than never tobacco users. CONCLUSIONS: Bidi is no less hazardous than cigarette smoking, and smokeless tobacco use may also result in significantly increased mortality. PMID- 16249219 TI - Three-year duration of benefit from abciximab in patients receiving stents for acute myocardial infarction in the randomized double-blind ADMIRAL study. AB - AIMS: The Abciximab Before Direct Angioplasty and Stenting in Myocardial Infarction Regarding Acute and Long-term Follow-up (ADMIRAL) study demonstrated that early inhibition of the platelet glycoprotein IIb/IIIa (GP IIb/IIIa) receptor with abciximab led to improved coronary patency, left ventricular function, and clinical outcomes. The current long-term follow-up study evaluated the durability of the positive outcomes. METHODS AND RESULTS: The randomized double-blind ADMIRAL trial enrolled 300 patients who received either abciximab plus stenting or placebo plus stenting for the treatment of ST-elevation myocardial infarction (STEMI). Abciximab (bolus of 0.25 mg/kg body weight, followed by 12 h infusion of 0.125 microg/kg per min) was administered to 149 patients, whereas 151 patients received placebo. Long-term follow-up was conducted in a blinded manner by either patient chart review or telephone interview. Long-term follow-up data were obtained on 288 patients (96%). After 3 years, using an intent-to-treat analysis, the outcome of all-cause mortality occurred in 9.1% of abciximab-treated patients when compared with 12.2% of placebo patients, absolute and relative risk reductions of 3.1 and 25%, respectively (P=0.36). Parallel Kaplan-Meier curves were observed for the cumulative incidence of death or re-infarction, which was reduced from 16.9% in the placebo group to 11.8% in the abciximab group, absolute and relative risk reductions of 5.1 and 30%, respectively (P=0.20). Rates of recurrent ischaemia were significantly reduced from 21.7 to 11.5% (P=0.05). CONCLUSION: Adjunctive abciximab to primary stenting for STEMI elicits favourable clinical outcomes with the same absolute risk reductions of hard clinical outcomes from 30 days up to 3 years of follow-up. PMID- 16249220 TI - Pulmonary valve replacement in patients with tetralogy of Fallot and pulmonary regurgitation: early surgery similar to optimal timing of surgery? PMID- 16249221 TI - Indication of long-term endothelial dysfunction after sirolimus-eluting stent implantation. AB - AIMS: Endothelial dysfunction has been related both to progression of atherosclerotic disease and to future cardiovascular events. We assessed local epicardial endothelial function 6 months after sirolimus-eluting stent (SES) or bare metal stent (BS) implantation. METHODS AND RESULTS: In 12 patients (seven SES, five BS), endothelium-dependent vasomotion of a coronary segment 15 mm in length, starting 2 mm distal to the stent, was assessed with quantitative coronary angiography immediately after the procedure and at 6 months follow-up, after intracoronary infusion of acetylcholine. Intravascular ultrasound (IVUS) was performed and coronary flow reserve (CFR) assessed in all patients. At follow up significant vasoconstriction was seen in SES (median 32% diameter reduction from baseline) but not in BS (median 2% reduction) patients after acetylcholine infusion (P=0.03 for SES vs. BS); endothelium-independent vasodilatation to nitrates did not differ significantly between groups (20% SES, 5% BS, P=0.14). IVUS revealed no late unhealed dissections and CFR was comparable between groups (SES 3.1 vs. BS 3.2, n.s.). CONCLUSION: SES implantation may have an adverse effect on local endothelium-dependent vasomotor responses compared with BS implantation at 6 months. Long-term clinical consequences of this observation are still unknown. PMID- 16249222 TI - The role of glycoprotein IIb/IIIa inhibitors in primary percutaneous coronary intervention for ST elevation myocardial infarction. PMID- 16249223 TI - Study of the role of functional variants of SLC22A4, RUNX1 and SUMO4 in systemic lupus erythematosus. AB - BACKGROUND: Functional polymorphisms of the solute carrier family 22, member 4 (SLC22A4), runt related transcription factor 1 (RUNX1) and small ubiquitin-like modifier 4 (SUMO4) genes have been shown to be associated with several autoimmune diseases. OBJECTIVE: To test the possible role of these variants in susceptibility to or severity of systemic lupus erythematosus (SLE), on the basis that common genetic bases are shared by autoimmune disorders. METHODS: 597 SLE patients and 987 healthy controls of white Spanish origin were studied. Two additional cohorts of 228 SLE patients from Sweden and 122 SLE patients from Colombia were included. A case-control association study was carried out with six single nucleotide polymorphisms (SNP) spanning the SLC22A4 gene, one SNP in RUNX1 gene, and one additional SNP in SUM04 gene. RESULTS: No significant differences were observed between SLE patients and healthy controls when comparing the distribution of the genotypes or alleles of any of the SLC22A4, RUNX1, or SUMO4 polymorphisms tested. Significant differences were found in the distribution of the SUMO4 genotypes and alleles among SLE patients with and without nephritis, but after multiple testing correction, the significance of the association was lost. The association of SUMO4 with nephritis could not be verified in two independent SLE cohorts from Sweden and Colombia. CONCLUSIONS: These results suggest that the SLC22A4, RUNX1, and SUMO4 polymorphisms analysed do not play a role in the susceptibility to or severity of SLE. PMID- 16249224 TI - Incidence of lymphoma in a large primary care derived cohort of cases of inflammatory polyarthritis. AB - OBJECTIVE: To determine the risk of lymphoma in a primary care derived cohort of new onset cases of inflammatory polyarthritis and assess the contribution of disease severity and standard immunosuppressive treatment. DESIGN: Prospective cohort study. METHODS: 2105 subjects with new onset inflammatory polyarthritis were recruited to the Norfolk Arthritis Register (NOAR) and followed annually for (median) 8.4 years. Occurrence of lymphoma was determined by annual morbidity review and linkage to the central hospital database serving the NOAR area. Cases of lymphoma were verified by record review. Standardised incidence ratios (SIRs) for lymphoma were calculated compared with the local, age, sex, and calendar year expected rates. Stratified analyses were undertaken for various markers of disease severity and treatment history. RESULTS: There were 11 cases of lymphoma during 15,548 person years of follow up, the majority of which were of large B cell type. Compared with the local population the SIR was 2.4 (95% confidence interval, 1.2 to 4.2). The risks in cases classified as rheumatoid arthritis, ever rheumatoid factor positive, or ever treated with DMARDs were all higher, the highest risk group being those treated with methotrexate: SIR = 4.9 (1.8 to 10.6). CONCLUSIONS: There was a doubling in risk of lymphoma in new onset cases of inflammatory polyarthritis. Patients with the most severe disease were twice as likely as other patients to develop lymphoma. These results need to be taken into account when considering reported increased risks of lymphoma compared to background population risk in users of new biological agents. PMID- 16249225 TI - Prevalence and predictors of disability in valued life activities among individuals with rheumatoid arthritis. AB - OBJECTIVE: To identify the prevalence of disability in a wide range of life activities and identify factors associated with such disability using the Verbrugge and Jette disablement model as a framework. METHODS: Data were from a panel study of 548 individuals with rheumatoid arthritis, interviewed annually by telephone. Valued life activity (VLA) disability was assessed using a 26-item scale rating difficulty in carrying out each activity. Three types of summary measure were calculated: activities unable to perform, activities affected, and mean difficulty. Subscale scores were also calculated, corresponding to obligatory, committed, and discretionary activities, as defined in the disablement model. Disease status measures were examined as predictors of VLA disability using multiple regression analyses. RESULTS: Half the subjects were unable to do at least one VLA. Approximately 2%, 31.3%, and 40.2% were unable to do at least one obligatory, committed, and discretionary activity, respectively. Almost all (95%) reported at least one VLA affected by rheumatoid arthritis; 68.4%, 91.4%, and 92.5% reported at least one obligatory, committed, and discretionary activity, respectively, affected. Disease status measures were robust predictors of VLA disability, accounting for 22-47% of the variation in VLA disability (with one exception). Adding the health assessment questionnaire (HAQ) to these models increased (p<0.0001) all model R2 values. HAQ score mediated the effects of many disease measures, consistent with the disablement model. CONCLUSION: VLA disability was common, with more disability noted in committed and discretionary than obligatory activities. Because VLA disability has been linked to psychological wellbeing in previous studies, identification of factors that may protect against such disability is important. PMID- 16249226 TI - PD-0200347, an alpha2delta ligand of the voltage gated calcium channel, inhibits in vivo activation of the Erk1/2 pathway in osteoarthritic chondrocytes: a PKCalpha dependent effect. AB - OBJECTIVE: To explore the in vivo effects of PD-0200347, an alpha(2)delta ligand of voltage gated Ca(2+) channels, on cell signalling in osteoarthritic (OA) chondrocytes from an experimental dog model, and examine the effect of PD-0200347 on the major signalling pathways involved in OA cartilage degradation. METHODS: OA was surgically induced in dogs by sectioning the anterior cruciate ligament. OA dogs were divided into three groups and treated orally with (a) placebo; (b) 15 mg/kg/day PD-0200347, or (c) 90 mg/kg/day PD-0200347. The animals were killed 12 weeks after surgery. Cartilage specimens from femoral condyles and tibial plateaus were processed for immunohistochemistry. Specific antibodies against the phosphorylated form of PKCalpha, Ras, c-Raf, the MAP kinases Erk1/2, p38, JNK, and the transcription factors, CREB and Elk-1, were used. RESULTS: Levels of all the tested signalling mediators were increased in the placebo treated (OA) group compared with the normal group. PD-0200347 treatment significantly reduced the levels of the active forms of PKCalpha, c-Raf, Erk1/2, and Elk-1; however, the levels of the active forms of Ras, p38, JNK, and CREB were not affected by the PD 0200347 treatment. CONCLUSION: The action of PD-0200347 on OA chondrocytes is probably mediated through the inhibition of Erk1/2 activation via a Ras independent mechanism. This effect is associated with reduction of the activation of transcription factors such as Elk-1, which leads to the inhibition of the induction of the major catabolic factors involved in the degradation process of OA cartilage. PMID- 16249227 TI - Anchorage on fibronectin via VLA-5 (alpha5beta1 integrin) protects rheumatoid synovial cells from Fas-induced apoptosis. AB - BACKGROUND: Rheumatoid synovial cells are resistant to apoptosis induction in vivo, whereas, fibroblast-like synovial cells in rheumatoid arthritis (RA-FLS) are vulnerable to Fas-induced apoptosis in vitro. OBJECTIVE: To clarify this discrepancy by studying the contribution of the interaction between cellular integrin and matrix fibronectin (Fn), which is significantly increased in the rheumatoid joints, to the induction of apoptosis in RA-FLS. METHODS: Integrin and Fas mRNAs were measured by reverse transcription-polymerase chain reaction in RA FLS. Integrins expressed in rheumatoid synovial tissues were analysed by immunohistochemistry. RA-FLS plated either on Fn or on control poly-L-lysine were incubated with agonistic anti-Fas monoclonal antibodies (mAbs). Apoptosis induction was evaluated using terminal deoxynucleotidyl transferase mediated UTP nick end labelling (TUNEL) and immunoblotting for caspase-3 and poly (ADP-ribose) polymerase in the presence or absence of anti-VLA-5 mAb. RESULTS: VLA-5 (alpha5beta1 integrin), a major integrin expressed on RA-FLS, was required for the adhesion of RA-FLS on Fn. RA-FLS plated on Fn were more resistant to Fas induced apoptosis than those plated on control poly-L-lysine. This protection by Fn was reversed by anti-VLA-5 mAb. CONCLUSION: Anchorage of RA-FLS on matrix Fn via VLA-5 protects RA-FLS from Fas-induced apoptosis, and Fn abundantly present in rheumatoid synovium appears to afford RA-FLS resistance against apoptosis induction in vivo. PMID- 16249228 TI - Finnish HLA studies confirm the increased risk conferred by HLA-B27 homozygosity in ankylosing spondylitis. AB - OBJECTIVE: To determine the influence of HLA-B27 homozygosity and HLA-DRB1 alleles in the susceptibility to, and severity of, ankylosing spondylitis in a Finnish population. METHODS: 673 individuals from 261 families with ankylosing spondylitis were genotyped for HLA-DRB1 alleles and HLA-B27 heterozygosity/homozygosity. The frequencies of HLA-B27 homozygotes in probands from these families were compared with the expected number of HLA-B27 homozygotes in controls under Hardy-Weinberg equilibrium (HWE). The effect of HLA-DRB1 alleles was assessed using a logistic regression procedure conditioned on HLA-B27 and case-control analysis. RESULTS: HLA-B27 was detected in 93% of cases of ankylosing spondylitis. An overrepresentation of HLA-B27 homozygotes was noted in ankylosing spondylitis (11%) compared with the expected number of HLA-B27 homozygotes under HWE (4%) (odds ratio (OR) = 3.3 (95% confidence interval, 1.6 to 6.8), p = 0.002). HLA-B27 homozygosity was marginally associated with reduced BASDAI (HLA-B27 homozygotes, 4.5 (1.6); HLA-B27 heterozygotes, 5.4 (1.8) (mean (SD)), p = 0.05). Acute anterior uveitis (AAU) was present in significantly more HLA-B27 positive cases (50%) than HLA-B27 negative cases (16%) (OR = 5.4 (1.7 to 17), p<0.004). HLA-B27 positive cases had a lower average age of symptom onset (26.7 (8.0) years) compared with HLA-B27 negative cases (35.7 (11.2) years) (p<0.0001). CONCLUSIONS: HLA-B27 homozygosity is associated with a moderately increased risk of ankylosing spondylitis compared with HLA-B27 heterozygosity. HLA-B27 positive cases had an earlier age of onset of ankylosing spondylitis than HLA-B27 negative cases and were more likely to develop AAU. HLA-DRB1 alleles may influence the age of symptom onset of ankylosing spondylitis. PMID- 16249229 TI - Impact of parental history on patients' cardiovascular mortality in rheumatoid arthritis. AB - BACKGROUND: Patients with rheumatoid arthritis are at increased risk of death from cardiovascular disease (CVD). This risk is influenced by the inflammatory activity of the rheumatoid arthritis as well as by traditional risk factors for CVD. However, little is known about whether or to what extent hereditary factors for CVD contribute additional risk in patients with rheumatoid arthritis. OBJECTIVE: To assess the clinical impact of a parental history of CVD in patients with rheumatoid arthritis. METHODS: Population based cohort study of 10,805 Swedish patients with rheumatoid arthritis aged 16-67 years during follow up (1990-2000). Parents, and cardiovascular deaths among patients and parents, were identified through register linkages. Relative risk of death v the general population was assessed using standardised mortality ratios (SMR), which were compared by Poisson regression. RESULTS: Rheumatoid patients with a parental history of fatal CVD had an SMR of death from CVD of 2.9 (95% confidence interval, 2.5 to 3.4). By contrast, rheumatoid patients without a parental history of fatal CVD had an SMR of 1.7 (1.2 to 2.3). A parental death from CVD was associated with a 70% increase in the risk of fatal CVD in rheumatoid arthritis (SMR ratio = 1.7 (1.2 to 2.4), and an increase in the 10 year mortality from CVD from 5% to 10% in men and from 2% to 4% in women aged 50 to 67 years. CONCLUSIONS: Parental history of death from CVD is an important (and easily assessable) risk factor for fatal CVD in rheumatoid arthritis. PMID- 16249230 TI - Antibodies against human 60 kDa heat shock protein are not associated with cardiovascular disease in patients with rheumatoid arthritis. AB - BACKGROUND: Rheumatoid arthritis is associated with an unexplained increased risk of cardiovascular disease (CVD). Antibodies against human 60 kDa heat shock protein (anti-HSP60) are associated with the presence and severity of CVD. OBJECTIVES: To investigate whether anti-HSP60 antibodies are associated with prevalent CVD in patients with rheumatoid arthritis. METHODS: In a nested case control design, anti-HSP60 antibody levels were measured in the serum samples of 192 rheumatoid patients. In a regression analysis the association between prevalent CVD and anti-HSP60 antibodies was examined, along with the possible influence on this association of several demographic, rheumatoid arthritis, and CVD related variables. RESULTS: In a random sample of 326 patients with rheumatoid arthritis, 48 cases were identified who also suffered from CVD. Three controls per case with rheumatoid arthritis but without CVD (n = 144) were matched for sex, age, disease duration, and smoking habits. A regression analysis showed no significant association between prevalent CVD and anti-HSP60 antibodies (odds ratio = 1.00 (95% confidence interval, 0.997 to 1.004)). After correcting for possible confounding variables, still no association was found. CONCLUSIONS: In contrast to the general population, anti-HSP60 antibody titres are not associated with prevalent CVD in patients with rheumatoid arthritis. These findings could be the result of an altered immune response to HSP60 in rheumatoid arthritis. PMID- 16249232 TI - Proteasomes degrade proteins in focal subdomains of the human cell nucleus. AB - The ubiquitin proteasome system plays a fundamental role in the regulation of cellular processes by degradation of endogenous proteins. Proteasomes are localized in both, the cytoplasm and the cell nucleus, however, little is known about nuclear proteolysis. Here, fluorogenic precursor substrates enabled detection of proteasomal activity in nucleoplasmic cell fractions (turnover 0.0541 microM/minute) and nuclei of living cells (turnover 0.0472 microM/minute). By contrast, cell fractions of nucleoli or nuclear envelopes did not contain proteasomal activity. Microinjection of ectopic fluorogenic protein DQ-ovalbumin revealed that proteasomal protein degradation occurs in distinct nucleoplasmic foci, which partially overlap with signature proteins of subnuclear domains, such as splicing speckles or promyelocytic leukemia bodies, ubiquitin, nucleoplasmic proteasomes and RNA polymerase II. Our results establish proteasomal proteolysis as an intrinsic function of the cell nucleus. PMID- 16249231 TI - Immunohistological examination of open sacroiliac biopsies of patients with ankylosing spondylitis: detection of tumour necrosis factor alpha in two patients with early disease and transforming growth factor beta in three more advanced cases. AB - OBJECTIVE: To characterise the immunohistological features of sacroiliitis in ankylosing spondylitis (AS) at different disease stages. METHODS: Biopsy samples from sacroiliac joints (SIJs) of five patients with AS, two with early, three with advanced changes and samples from age matched controls from one necropsy SIJ and two iliac bone marrow (BM) biopsies were studied. Paraffin sections were immunostained in triplicate for T cells (CD3, CD8), macrophages (CD68), and the cytokines tumour necrosis factor alpha (TNFalpha), interferon gamma, interleukin (IL) 1beta, IL6, IL10, and transforming growth factor beta1 (TGFbeta1). Stained cells were counted over one entire high power field (x400) per section in BM, cartilage, and other connective tissue (CT). Results are the mean of three sections. RESULTS: CD3+ T cells were numerous in the BM of early AS, and in the CT of one patient with early and one with late AS, with variable proportions of CD8+ T cells. All patients with AS had more CD68+ macrophages than controls in BM and CT; in cartilage, one patient with early and one with late AS had increased CD68+ cells, some being osteoclasts. The patient with very early AS had large numbers of TNFalpha cells in the three tissular areas; for the other patient with early disease they were found only in CT and cartilage. IL6 was seen in 4/4 patients with AS in most areas. Patients with early disease had more T cells, TNFalpha, and IL6, and patients with advanced AS more TGFbeta1. CONCLUSION: The immunohistological findings of a limited sample suggest a role for BM in sacroiliitis, for TNFalpha and IL6 in early, active lesions, and for TGFbeta1 at the time of secondary cartilage and bone proliferation. PMID- 16249233 TI - Distribution and dynamics of Lamp1-containing endocytic organelles in fibroblasts deficient in BLOC-3. AB - Late endosomes and lysosomes of mammalian cells in interphase tend to concentrate in the perinuclear region that harbors the microtubule-organizing center. We have previously reported abnormal distribution of these organelles - as judged by reduced percentages of cells displaying pronounced perinuclear accumulation - in mutant fibroblasts lacking BLOC-3 (for ;biogenesis of lysosome-related organelles complex 3'). BLOC-3 is a protein complex that contains the products of the genes mutated in Hermansky-Pudlak syndrome types 1 and 4. Here, we developed a method based on image analysis to estimate the extent of organelle clustering in the perinuclear region of cultured cells. Using this method, we corroborated that the perinuclear clustering of late endocytic organelles containing Lamp1 (for ;lysosome-associated membrane protein 1') is reduced in BLOC-3-deficient murine fibroblasts, and found that it is apparently normal in fibroblasts deficient in BLOC-1 or BLOC-2, which are another two protein complexes associated with Hermansky-Pudlak syndrome. Wild-type and mutant fibroblasts were transfected to express human LAMP1 fused at its cytoplasmic tail to green fluorescence protein (GFP). At low expression levels, LAMP1-GFP was targeted correctly to late endocytic organelles in both wild-type and mutant cells. High levels of LAMP1-GFP overexpression elicited aberrant aggregation of late endocytic organelles, a phenomenon that probably involved formation of anti-parallel dimers of LAMP1-GFP as it was not observed in cells expressing comparable levels of a non-dimerizing mutant variant, LAMP1-mGFP. To test whether BLOC-3 plays a role in the movement of late endocytic organelles, time-lapse fluorescence microscopy experiments were performed using live cells expressing low levels of LAMP1-GFP or LAMP1-mGFP. Although active movement of late endocytic organelles was observed in both wild type and mutant fibroblasts, quantitative analyses revealed a relatively lower frequency of microtubule-dependent movement events, either towards or away from the perinuclear region, within BLOC-3-deficient cells. By contrast, neither the duration nor the speed of these microtubule-dependent events seemed to be affected by the lack of BLOC-3 function. These results suggest that BLOC-3 function is required, directly or indirectly, for optimal attachment of late endocytic organelles to microtubule-dependent motors. PMID- 16249234 TI - Fundamentally different roles for LFA-1, Mac-1 and alpha4-integrin in neutrophil chemotaxis. AB - Although the LFA-1, Mac-1 and alpha(4) integrins are required for chemotaxis, it is unknown how they are regulated or what specific role they play. Previously we demonstrated that fMLP and IL-8 induce chemotaxis via the p38 MAPK and phosphoinositide 3-kinase (PI3K) pathways, respectively. Here we show that these chemoattractants also activate and use Mac-1 and LFA-1 in a differential manner during chemotaxis. Using integrin-specific substrata, we demonstrate that cell movement in response to IL-8 is mediated by Mac-1, whereas LFA-1 is required for directional migration. By contrast, chemotaxis to fMLP requires Mac-1 for cell movement, whereas LFA-1 and alpha(4)-integrin are required for directional migration. On serum protein, which contains ligands for LFA-1, Mac-1 and alpha(4) integrin, chemotaxis to fMLP is dependent on Mac-1, whereas chemotaxis to IL-8 is dependent on LFA-1. These results suggest that Mac-1 is the dominant integrin involved in chemotaxis to fMLP, and LFA-1 is the dominant integrin involved in chemotaxis to IL-8. Consistent with these observations, higher quantities of high affinity Mac-1 are found on cells chemotaxing to fMLP then on cells chemotaxing to IL-8. Moreover, a much larger quantity of clustered LFA-1 was found on cells migrating to IL-8 compared to cells moving towards fMLP. When cells are presented with competing gradients of fMLP and IL-8, they preferentially migrate towards fMLP and activate/utilize integrins in a manner identical to fMLP alone. Under the same conditions, p38 MAPK inhibition abolishes the preferential migration to fMLP; instead, the cells migrate preferentially towards IL-8. The activation and utilization of integrins under these conditions are consistent with patterns observed with IL-8 alone. Together, these data suggest that fMLP and IL-8 differentially activate integrins for use during chemotaxis, that p38 MAPK is a major mediator in the activation and utilization of integrins, and selective integrin activation occurs during chemotaxis between opposing gradients. PMID- 16249235 TI - Interferon-gamma selectively increases epithelial permeability to large molecules by activating different populations of paracellular pores. AB - Impairment of the gut epithelial barrier by agents such as IFNgamma may play a key role in the pathogenesis of inflammatory disorders by increasing the paracellular penetration of luminal macromolecules, potentially including bacterial antigens. Owing to limitations of current paracellular probes, little is known about the precise functional changes induced by IFNgamma and how these relate to the development of increased macromolecular permeability. Here we investigate how IFNgamma modulates this pathway in T84 monolayers using a novel profiling technique that resolves different populations of paracellular pores by simultaneous analysis of 24 permeability probes of defined molecular size. Two types of functional pore present in control monolayers, an abundant restrictive pore with a radius of approximately 4.5 Angstrom and a much larger but infrequent, non-restrictive pore, were differentially regulated by IFNgamma. Incubation with IFNgamma dose-dependently and reversibly increased the frequency of the non-restrictive pores while having no significant effect on the restrictive component. Cytokine-induced increases in beta, the descriptor of the non-restrictive pore, correlated closely with increased permeability to large molecules (10 kDa) including E. coli-derived lipopolysaccharide, but not small (0.182 kDa) molecules. This effect was associated with changes in expression of the tight junction proteins occludin and claudin-1. These data suggest that IFNgamma selectively increases the transepithelial flux of large molecules by activating specific pathways within the junctional pore. One hypothesis is that this process may be activated in the early stages of the inflammatory response, facilitating the passage of large and potentially antigenic molecules across the gut without gross disruption of the barrier to small molecules. PMID- 16249236 TI - Shroom regulates epithelial cell shape via the apical positioning of an actomyosin network. AB - The actin-binding protein Shroom is essential for neural tube morphogenesis in multiple vertebrate organisms, indicating its function is evolutionarily conserved. Shroom facilitates neurulation by regulating the morphology of neurepithelial cells. Shroom localizes to the apical tip of adherens junctions of neural ectoderm cells in vivo and to the apical junctional complex (AJC) in MDCK cells. Induced expression of Shroom in polarized epithelia elicits apical constriction and dramatic reorganization of the apical arrangement and packing of cells without altering apical-basal polarity. These events likely mimic the cell shape changes and cellular movements required for neurulation in vivo. The observed phenotypes depend on the ability of Shroom to alter F-actin distribution and regulate the formation of a previously uncharacterized contractile actomyosin network associated with the AJC. Targeting the C-terminal domain of Shroom to the apical plasma membrane elicits constriction and reorganization of the actomyosin network, indicting that this domain mediates Shroom's activity. In vivo, Shroom mutant neural epithelia show a marked reduction in apically positioned myosin. Thus, Shroom likely facilitates neural tube closure by regulating cell shape changes via the apical positioning of an actomyosin network in the neurepithelium. PMID- 16249237 TI - Acetylcholine and calcium signalling regulates muscle fibre formation in the zebrafish embryo. AB - Nerve activity is known to be an important regulator of muscle phenotype in the adult, but its contribution to muscle development during embryogenesis remains unresolved. We used the zebrafish embryo and in vivo imaging approaches to address the role of activity-generated signals, acetylcholine and intracellular calcium, in vertebrate slow muscle development. We show that acetylcholine drives initial muscle contraction and embryonic movement via release of intracellular calcium from ryanodine receptors. Inhibition of this activity-dependent pathway at the level of the acetylcholine receptor or ryanodine receptor did not disrupt slow fibre number, elongation or migration but affected myofibril organisation. In mutants lacking functional acetylcholine receptors myofibre length increased and sarcomere length decreased significantly. We propose that calcium is acting via the cytoskeleton to regulate myofibril organisation. Within a myofibre, sarcomere length and number are the key parameters regulating force generation; hence our findings imply a critical role for nerve-mediated calcium signals in the formation of physiologically functional muscle units during development. PMID- 16249238 TI - Effects of loss of metatarsophalangeal joint mobility on gait in rheumatoid arthritis patients. AB - OBJECTIVE: To evaluate the effects of loss of range of motion (ROM) of the metatarsophalangeal (MTP) joint on the kinematic parameters of walking in rheumatoid arthritis (RA) patients. METHODS: Inclusion of RA patients with inactive disease, no synovitis of the inferior limb and reduced ROM of the MTP joints. Evaluation of the ROM of the MTP dorsal and plantar flexion, and gait analysis using a three-dimensional computerized movement analysis. Calculation of gait parameters and maximal flexion and extension of the hips and knees during walking. Analysis 1 compared the ROM of dorsal and plantar flexion in patients with or without walking pain; 2 compared the gait parameters between patients and controls; 3 investigated a relationship between gait parameters and (i) the ROM of the MTP dorsal and plantar flexion and (ii) the pain at walking; 4 investigated the relationship between the ROM of the MTP dorsal and plantar flexion and maximal flexion and extension of the hip and knee joints during walking. RESULTS: Nine patients and seven controls were included. The MTP ROM was no different in patients presenting with or without pain at walking. The walking velocity was lower and the stride length shorter in patients than in controls. The walking velocity and the stride length were positively related to the MTP dorsal flexion ROM (r(2)=0.75 and 0.67). There was a negative relationship between maximal flexion of the knee and hips during walking and the underlying MTP dorsal flexion ROM (r(2)=0.67 and 0.54). CONCLUSION: In RA patients, reduced MTP dorsal flexion mobility induces changes in the walking parameters, including the kinematics of the overlying lower limb joints. Treatment of an RA-impaired forefoot should focus on MTP mobility as well as on pain. PMID- 16249239 TI - The use of patient partners with back pain to teach undergraduate medical students. AB - OBJECTIVES: To assess the impact of teaching about back pain to medical students using trained patient partners (PP). METHODS: An initial training programme for four PPs (two with sciatica and two with ankylosing spondylitis) followed by teaching to alternate groups of medical students at the Whittington Campus of the Royal Free and University College Medical School (RFUCMS). A control group of students did not receive the PP teaching. All students received standard Whittington Campus rheumatology teaching. Performance in an end of year objective structured clinical examination (OSCE) was compared between the two groups. Student and PP perceptions of the teaching and training were evaluated using focus groups and questionnaires. RESULTS: Students receiving the PP teaching performed significantly better in a summative OSCE, but no difference was seen in analysis of a single station assessing history-taking skills in a patient with back pain. Students felt that the PP teaching improved their ability to elicit information from a patient during the consultation. PPs enjoyed the experience of teaching and felt empowered to self-manage their medical conditions, and were better able to seek medical advice when needed. CONCLUSIONS: Using PPs with back pain to teach medical students has a positive effect on student learning and patient well-being. The feasibility of delivering this programme will depend on faculty resources. The effects on examination performance are small but significant. PMID- 16249240 TI - Sir George Frederick Still (1868-1941). PMID- 16249241 TI - The usefulness of bone remodelling markers in predicting the efficacy of pamidronate treatment in SAPHO syndrome. AB - OBJECTIVES: Pamidronate has recently been used in SAPHO syndrome due to its anti osteoclastic effect. The aim of this study is to determine the usefulness of bone remodelling markers for determining the efficacy of pamidronate treatment. METHODS: Thirteen patients with SAPHO syndrome were treated with pamidronate. The treatment evaluation was done using a visual analogue scale (VAS) and also erythrocyte sedimentation rate, C-reactive protein, serum crosslaps (sCTX) and osteocalcin initially and after 3 months. A relevant clinical response was defined as an improvement in VAS of at least 40%. RESULTS: At 3 months, 7 of 13 patients had a good clinical response, as previously defined. Five of the seven patients maintained the good response over 6 months. Before the first perfusion 6 of the 13 patients had increased sCTX (upper 3250 pmol/l). In this small cohort we tried to analyse whether the increase in bone remodelling markers was associated with a good clinical response. In the responders group the mean levels of sCTX and osteocalcin at baseline were 6783.17 and 24.66, respectively, and in the non-responders group the levels were 2152 and 11.8, respectively. There was a significant difference in sCTX between the responders and the non-responders (P = 0.0044). CONCLUSION: Infusion of pamidronate is effective in SAPHO in some patients. Increased sCTX might be a prognostic marker for a good clinical response but results have to be confirmed in a larger cohort. PMID- 16249242 TI - The second trimester Doppler ultrasound examination is the best predictor of late pregnancy outcome in systemic lupus erythematosus and/or the antiphospholipid syndrome. AB - OBJECTIVE: To examine the predictive value of clinical examination, laboratory tests and Doppler ultrasound examination in systemic lupus erythematosus (SLE) and/or antiphospholipid syndrome (APS) pregnancies. METHODS: A prospective study of 116 pregnancies followed in a single tertiary referral centre. Outcomes analysed were fetal/neonatal death and adverse pregnancy outcome. Univariate analysis was performed for: (i) medical and obstetric history; (ii) medical and obstetric clinical examination; (iii) biological data; (iv) Doppler ultrasound examination. Variables significantly associated with the outcomes in the univariate analysis were entered into a logistic regression model. RESULTS: Sixteen out of 116 pregnancies ended in 12 fetal deaths and 4 embryonic losses. Hence, data for 100 pregnancies were analysed. Seven fetal deaths and one neonatal death occurred, associated with abnormal end-diastolic umbilical artery Doppler flow at the second trimester (P < 0.006), a history of thrombophlebitis (P < 0.001) or notched uterine artery and growth restriction at the second trimester (P < 0.002). Multivariate analysis retained abnormal end-diastolic umbilical artery Doppler flow (P = 0.047) and history of thrombophlebitis (P = 0.018) as significant predictors. Thirty-one adverse pregnancy outcomes occurred, associated with notched uterine artery (P < 0.00003), abnormal end-diastolic umbilical artery Doppler flow (P < 0.0006) and fetal growth restriction at the second trimester (P < 0.008), growth restriction (P < 0.00001) and notched uterine artery at the third trimester (P < 0.0008), use of heparin (P < 0.05) and history of thrombophlebitis (P < 0.04). Notched uterine artery at the second trimester remained the only predictor in multivariate analysis (P = 0.001). CONCLUSIONS: Results of the second trimester Doppler ultrasound examination are the best predictors for late pregnancy outcome in SLE and/or APS. PMID- 16249243 TI - Anti-inflammatory effects of contralateral administration of the kappa-opioid agonist U-50,488H in rats with unilaterally induced adjuvant arthritis. AB - OBJECTIVE: The effect of repeated contralateral treatment with the kappa-opioid receptor agonist U-50,488H {trans-(+/-)-3,4-dichloro-N-methyl-N-[2-(1 pyrrolidinyl)-cyclohexyl]-benzene acetamide methanesulphonate} was investigated in rats with unilaterally induced adjuvant arthritis. METHODS: Arthritis was induced by injection of Mycobacterium butyricum into the right hindpaw. Inflammatory parameters, nociceptive behaviour and cartilage turnover were evaluated up to 21 days after induction of arthritis. RESULTS: Contralateral treatment with 0.3 mg U-50,488H into the left hindpaw twice per week reduced the hindpaw oedema, ankle joint inflammation, pain behaviour to mechanical stimuli and severity score of inflammation in the hindpaws of both sides as well as the systemic spread of inflammation to other areas, e.g. tail and/or forepaws, compared with saline-treated animals. Moreover, a significant decrease in the levels of cartilage oligomeric matrix protein was found in animals treated with U 50,488H, suggesting reduction of cartilage damage. The anti-inflammatory and chondroprotective effects of U-50,488H were abolished by administration of the peripheral opioid receptor antagonist naloxone methiodide. CONCLUSIONS: This is the first report demonstrating that repeated contralateral administration of a kappa-opioid receptor agonist diminishes the development of a symmetrical joint disorder. PMID- 16249244 TI - A statistical analysis of the interrelationships between disease activity in different systems in systemic lupus erythematosus. AB - OBJECTIVES: To develop models for disease activity in patients with systemic lupus erythematosus (SLE) and to examine the hypothesis that possible subsets exist within the disease, notably renal disease and little else, mucocutaneous and musculoskeletal disease in isolation and more multisystem disease. METHODS: Four hundred and forty patients with SLE were followed for a period of 10 yr. Socio-demographic data were obtained at the first visit with disease activity being recorded at subsequent visits and damage scores at 6-monthly intervals. Prognostic factors for active disease in each of the mucocutaneous, musculoskeletal and renal systems were examined statistically. The results were then validated using data collected over 5 yr on a further 295 SLE patients from a different centre. RESULTS: Logistic regression analyses indicated that for all three systems studied a patient known to have an involvement in that system is more likely to present with active disease in that same system than a patient with no known prior involvement. Patients with a higher frequency of clinic visits with active disease in a system are more likely to represent with active disease than those with fewer visits. The results suggest that renal disease is most likely to occur on its own. Associations between activity in the mucocutaneous and musculoskeletal systems support the suggestion that patients with musculoskeletal and mucocutaneous disease alone represent a possible subset of SLE. None of the associations identified were modified by the medication a patient received. CONCLUSIONS: Previous disease history and involvement of other systems determine a patient's chance of developing further episodes of active disease in SLE. PMID- 16249245 TI - Abnormal apoptosis in chronic granulomatous disease and autoantibody production characteristic of lupus. AB - OBJECTIVES: Patients with chronic granulomatous disease and carrier mothers of patients with chronic granulomatous disease are predisposed to developing various forms of lupus. This disorder is a neutrophil defect in intracellular killing. Abnormal apoptosis has been described. We hypothesized that abnormal apoptosis occurring in neutrophils of patients made them more immunogenic. METHODS: Human patients with chronic granulomatous disease were examined for abnormalities of neutrophil apoptosis by flow cytometry. To model the effect of abnormal apoptosis, a murine model was used. Apoptotic cells from either wild type or mice with chronic granulomatous disease were injected into either wild type or chronic granulomatous disease mice and autoantibodies were determined by ELISA. RESULTS: Our studies found that human and murine neutrophils carrying the gp91 form of chronic granulomatous disease had impaired exposure of phosphatidyl serine on the surface. Other markers of apoptosis were largely normal. Injection of apoptotic neutrophils from gp91 knockout mice into gp91 knockout mice led to the development of characteristic autoantibodies of lupus. CONCLUSIONS: Humans with chronic granulomatous disease may be at an increased risk of developing lupus due to abnormal apoptosis and abnormal clearance of apoptotic cells. PMID- 16249247 TI - Serum free prolactin concentrations in patients with systemic lupus erythematosus are associated with lupus activity. AB - OBJECTIVE: To determine in patients with systemic lupus erythematosus (SLE) the relationships among serum total and free prolactin (PRL) levels, isoforms of PRL and lupus activity. METHODS: In a cross-sectional study, 259 patients with SLE were tested for serum total PRL, serum free PRL, and PRL in fractions obtained after gel filtration chromatography (in 70 sera taken at random) by immunoradiometric assay based on disease activity. RESULTS: A significant correlation between direct PRL and free PRL levels was found in patients with and without lupus activity (r = 0.475, P<0.001); however, this was less so for non active patients than for active patients (r=0.433, P<0.001 and r=0.909, P<0.001, respectively). SLE Disease Activity Index (SLEDAI) scores correlated positively with serum free PRL levels (r=0.314, P<0.001) and percentage of little PRL (r=0.33, P=0.005) and negatively with percentage of big big PRL (r=-0.3, P=0.012). Patients with active disease had higher serum free PRL levels (median 12.6 vs 9.3 ng/ml, P<0.001), higher percentages of little PRL (83.1 +/- 21.2 vs 63.6 +/- 24.8%, P=0.011) and lower percentages of big big PRL (9.4 +/- 18.0 vs 25.2 +/- 24.3%, P=0.031). Different clinical manifestations and serological parameters of lupus disease activity were more frequent in patients with free hyperprolactinaemia than in patients with normal serum free PRL levels (such as neurological manifestations, renal involvement, serositis, haematological manifestations, anti-double-stranded DNA and hypocomplementaemia; P<0.021). CONCLUSION: An increase in serum free PRL levels, higher percentages of little PRL and lower percentages of big big PRL proved to be factors related to lupus activity in a subset of patients with SLE. These novel data must be taken into account in future studies aiming to establish a relationship between PRL and disease activity in SLE. PMID- 16249246 TI - Curcumin synergistically potentiates the growth-inhibitory and pro-apoptotic effects of celecoxib in osteoarthritis synovial adherent cells. AB - OBJECTIVES: Osteoarthritis (OA) is the Western world's leading cause of disability. Cyclo-oxygenase-2 (COX-2) inhibitors are efficient anti-inflammatory agents commonly used in the treatment of osteoarthritis. However, recent studies have shown that their long-term use may be limited due to cardiovascular toxicity. The anti-inflammatory efficacy of the phytochemical curcumin has been demonstrated in several in vitro and animal models. This study was undertaken to investigate whether curcumin augments the growth-inhibitory and pro-apoptotic effects of celecoxib in OA synovial adherent cells. METHODS: OA synovial adherent cells were prepared from human synovial tissue collected during total knee replacement surgery. The cells were exposed to different concentrations of celecoxib (0-40 mum), curcumin (0-20 mum) and their combination. Flow cytometric analysis was used to measure the percentage of cells with a subdiploid DNA content, the hallmark of apoptosis. COX-2 activity was assessed by measuring production of prostaglandin E(2) by enzyme-linked immunoassay. RESULTS: A synergistic effect was observed in inhibition of cell growth when the cells were exposed to various concentrations of celecoxib combined with curcumin. The inhibitory effect of the combination on cell growth was associated with an increased induction of apoptosis. The synergistic effect was mediated through a mechanism that involves inhibition of COX-2 activity. CONCLUSIONS: This effect may enable the use of celecoxib at lower and safer concentrations, and may pave the way for a novel combination treatment in osteoarthritis and other rheumatological disorders. PMID- 16249248 TI - Physician-owned specialty hospitals: a market signal for Medicare payment revisions. AB - Jean Mitchell's findings show that physician-entrepreneurs respond to financial incentives and take advantage of variations in profitability within Medicare's hospital payment system. The growth of physician-owned specialty hospitals can be seen as the reflection of parallel growth in profit opportunities. As Medicare plans to do, payments should be revised to squeeze out excess profits. Prohibiting physicians' use of hospitals they own might be unnecessary and could make it harder to identify future distortions in Medicare prices. If squeezing out excess profits threatens general hospitals' social missions, then new and explicit ways of identifying and funding social missions must be found. PMID- 16249249 TI - Effects of physician-owned limited-service hospitals: evidence from Arizona. AB - In recent years physician ownership of so-called limited-service hospitals has become commonplace in many states lacking certificate-of-need regulations. Empirical evidence documenting the effects of these facilities is sparse. This study compares practice patterns of physician-owners of limited-service cardiac hospitals and physician-nonowners who treat cardiac patients at competing full service community hospitals. Analyses of six years of Arizona inpatient discharge data show that physician-owners treat higher volumes of profitable cardiac surgical diagnosis-related groups (DRGs), higher percentages of low-severity cases, and higher percentages of cases with generous insurance compared with physician-nonowners who treat cardiac patients in community hospitals. PMID- 16249250 TI - The rise of the entrepreneurial physican. AB - The policy issues surrounding physician-owned specialty hospitals are highly controversial. Central to the controversy is the trade-off between the role these hospitals might play in increasing competition and the impact they might have on community hospitals' ability to cross-subsidize unfunded missions. Key policy questions relate to quality, efficiency, and the degree to which specialty hospitals are fairly paid for their services. This commentary reviews Jean Mitchell's basic thesis in relation to both the emerging specialty hospital literature and earlier work performed by the Lewin Group for MedCath, a corporation that owns and manages heart specialty hospitals. PMID- 16249251 TI - Low-intensity contraction activates the alpha1-isoform of 5'-AMP-activated protein kinase in rat skeletal muscle. AB - Skeletal muscle expresses two catalytic subunits, alpha1 and alpha2, of the 5' AMP-activated protein kinase (AMPK), which has been implicated in contraction stimulated glucose transport and fatty acid oxidation. Muscle contraction activates the alpha2-containing AMPK complex (AMPKalpha2), but this activation may occur with or without activation of the alpha1-containing AMPK complex (AMPKalpha1), suggesting that AMPKalpha2 is the major isoform responsible for contraction-induced metabolic events in skeletal muscle. We report for the first time that AMPKalpha1, but not AMPKalpha2, can be activated in contracting skeletal muscle. Rat epitrochlearis muscles were isolated and incubated in Krebs Ringer bicarbonate buffer containing pyruvate. In muscles stimulated to contract at a frequency of 1 and 2 Hz during the last 2 min of incubation, AMPKalpha1 activity increased twofold and AMPKalpha2 activity remained unchanged. Muscle stimulation did not change the muscle AMP concentration or the AMP-to-ATP ratio. AMPK activation was associated with increased phosphorylation of Thr(172) of the alpha-subunit, the primary activation site. Muscle stimulation increased the phosphorylation of acetyl-CoA carboxylase (ACC), a downstream target of AMPK, and the rate of 3-O-methyl-d-glucose transport. In contrast, increasing the frequency (>or=5 Hz) or duration (>or=5 min) of contraction activated AMPKalpha1 and AMPKalpha2 and increased AMP concentration and the AMP/ATP ratio. These results suggest that 1) AMPKalpha1 is the predominant isoform activated by AMP independent phosphorylation in low-intensity contracting muscle, 2) AMPKalpha2 is activated by an AMP-dependent mechanism in high-intensity contracting muscle, and 3) activation of each isoform enhances glucose transport and ACC phosphorylation in skeletal muscle. PMID- 16249252 TI - Insulin secretion in the conscious mouse is biphasic and pulsatile. AB - Islets in most species respond to increased glucose with biphasic insulin secretion, marked by a sharp first-phase peak and a slowly rising second phase. Mouse islets in vitro, however, lack a robust second phase. To date, this observation has not been extended in vivo. We thus compared insulin secretion from conscious mice with isolated mouse islets in vitro. The arterial plasma insulin response to a hyperglycemic clamp was measured in conscious mice 1 wk after surgical implantation of carotid artery and jugular vein catheters. Mice were transfused using clamps with blood from a donor mouse to maintain blood volume, allowing frequent arterial sampling. When plasma glucose in vivo was raised from approximately 5 to approximately 13 mM, insulin rose to a first-phase peak of 403+/-73% above basal secretion (n=5), followed by a rising second phase of mean 289+/- 41%. In contrast, perifused mouse islets ( approximately 75 islets/trial) responded with a similar first phase of 508+/- 94% (n=4) but a smaller and virtually flat second phase of 169+/- 9% (n=4, P<0.05). Furthermore, the slope of the second-phase response differed significantly from zero in mice (2.63+/-0.39%/min, P<0.01), in contrast to perifused islets (0.18+/- 0.14%/min, P>0.30). Mice also displayed pulsatile patterns in insulin concentration (period: 4.2+/- 0.4 min, n=8). Conscious mice thus responded to increased glucose with biphasic and pulsatile insulin secretion, as in other species. The robust second phase observed in vivo suggests that the processes needed to generate second phase insulin secretion may be abrogated by islet isolation. PMID- 16249253 TI - De novo emergence of insulin-stimulated glucose uptake in human aortic endothelial cells incubated with high glucose. AB - Elevated glucose concentrations have profound effects on cell function. We hypothesized that incubation of human aortic endothelial cells (HAEC) with high glucose increases insulin signaling and develops the appearance of insulin stimulated glucose uptake by the cells. Compared with 5 mM glucose, incubation of HAEC with 30 mM glucose for up to 48 h increased in a time-dependent manner expression of insulin receptor, insulin receptor substrate (IRS)-1, IRS-2, and GLUT1 proteins. High glucose also increased the specific binding of (125)I labeled insulin in HAEC accompanied by accelerated production of interleukin (IL) 6 and IL-8. Short-term stimulation by 50 microU/ml insulin did not activate [(14)C]glucose uptake by HAEC incubated in 5 mM glucose. However, an addition of insulin to high glucose-exposed endothelial cells led to a significant increase in [(14)C]glucose uptake in a glucose concentration- and time-dependent fashion, reaching a plateau at 48 h of incubation. Furthermore, incubation of HAEC with 30 mM glucose resulted in a new insulin-stimulated extracellular signal-regulated kinase-1/2 mitogen-activated protein kinase phosphorylation and increased lipid peroxidation and production of reactive oxygen species. These studies show for the first time that high glucose increases expression of insulin receptors and downstream elements of the insulin-signaling pathway and transforms "insulin resistant" aortic endothelial cells into "insulin-sensitive" tissue regarding glucose uptake. PMID- 16249254 TI - Glucose inhibits GABA release by pancreatic beta-cells through an increase in GABA shunt activity. AB - GABA is the major inhibitory neurotransmitter in the nervous system. It is also released by the insulin-producing beta-cells, providing them with a potential paracrine regulator. Because glucose was found to inhibit GABA release, we investigated whether extracellular GABA can serve as a marker for glucose-induced mitochondrial activity and thus for the functional state of beta-cells. GABA release by rat and human beta-cells was shown to reflect net GABA production, varying with the functional state of the cells. Net GABA production is the result of GABA formation through glutamate decarboxylase (GAD) and GABA catabolism involving a GABA-transferase (GABA-T)-mediated shunt to the TCA cycle. GABA-T exhibits K(m) values for GABA (1.25 mM) and for alpha-ketoglutarate (alpha-KG; 0.49 mM) that are, respectively, similar to and lower than those in brain. The GABA-T inhibitor gamma-vinyl GABA was used to assess the relative contribution of GABA formation and catabolism to net production and release. The nutrient status of the beta-cells was found to regulate both processes. Glutamine dose dependently increased GAD-mediated formation of GABA, whereas glucose metabolism shunts part of this GABA to mitochondrial catabolism, involving alpha-KG-induced activation of GABA-T. In absence of extracellular glutamine, glucose also contributed to GABA formation through aminotransferase generation of glutamate from alpha-KG; this stimulatory effect increased GABA release only when GABA-T activity was suppressed. We conclude that GABA release from beta-cells is regulated by glutamine and glucose. Glucose inhibits glutamine-driven GABA formation and release through increasing GABA-T shunt activity by its cellular metabolism. Our data indicate that GABA release by beta-cells can be used to monitor their metabolic responsiveness to glucose irrespective of their insulin secretory activity. PMID- 16249255 TI - Increased collagen content in insulin-resistant skeletal muscle. AB - Oversupply and underutilization of lipid fuels are widely recognized to be strongly associated with insulin resistance in skeletal muscle. Recent attention has focused on the mechanisms underlying this effect, and defects in mitochondrial function have emerged as a potential player in this scheme. Because evidence indicates that lipid oversupply can produce abnormalities in extracellular matrix composition and matrix changes can affect the function of mitochondria, the present study was undertaken to determine whether muscle from insulin-resistant, nondiabetic obese subjects and patients with type 2 diabetes mellitus had increased collagen content. Compared with lean control subjects, obese and type 2 diabetic subjects had reduced muscle glucose uptake (P<0.01) and decreased insulin stimulation of tyrosine phosphorylation of insulin receptor substrate-1 and its ability to associate with phosphatidylinositol 3-kinase (P<0.01 and P<.05). Because it was assayed by total hydroxyproline content, collagen abundance was increased in muscle from not only type 2 diabetic patients but also nondiabetic obese subjects (0.26+/-0.05, 0.57+/-0.18, and 0.67+/- 0.20 microg/mg muscle wet wt, lean controls, obese nondiabetics, and type 2 diabetics, respectively), indicating that hyperglycemia itself could not be responsible for this effect. Immunofluorescence staining of muscle biopsies indicated that there was increased abundance of types I and III collagen. We conclude that changes in the composition of the extracellular matrix are a general characteristic of insulin-resistant muscle. PMID- 16249256 TI - Whole body leucine flux in HIV-infected patients treated with or without protease inhibitors. AB - The present study was carried out to assess the effects of protease inhibitor (PI) therapy on basal whole body protein metabolism and its response to acute amino acid-glucose infusion in 14 human immunodeficiency virus (HIV)-infected patients. Patients treated with PIs (PI+, 7 patients) or without PIs (PI-, 7 patients) were studied after an overnight fast during a 180-min basal period followed by a 140-min period of amino acid-glucose infusion. Protein metabolism was investigated by a primed constant infusion of l-[1-(13)C]leucine. Dual-energy X-ray absorptiometry for determination of fat-free mass (FFM) and body fat mass measured body composition. In the postabsorptive state, whole body leucine balance was 2.5 times (P < 0.05) less negative in the PI+ than in the PI- group. In HIV-infected patients treated with PIs, the oxidative leucine disposal during an acute amino acid-glucose infusion was lower (0.58 +/- 0.09 vs. 0.81 +/- 0.07 micromol x kg FFM(-1) x min(-1) using plasma [(13)C]leucine enrichment, P = 0.06; or 0.70 +/- 0.10 vs. 0.99 +/- 0.08 micromol x kg FFM(-1) x min(-1) using plasma [(13)C]ketoisocaproic acid enrichment, P = 0.04 in PI+ and PI- groups, respectively) than in patients treated without PIs. Consequently, whole body nonoxidative leucine disposal (an index of protein synthesis) and leucine balance (0.50 +/- 0.10 vs. 0.18 +/- 0.06 micromol x kg FFM x (-1) x min(-1) in PI+ and PI groups respectively, P < 0.05) were significantly improved during amino acid glucose infusion in patients treated with PIs. However, whereas the response of whole body protein anabolism to an amino acid-glucose infusion was increased in HIV-infected patients treated with PIs, any improvement in lean body mass was detected. PMID- 16249257 TI - Dual mechanism of the potentiation by glucose of insulin secretion induced by arginine and tolbutamide in mouse islets. AB - Glucose induces insulin secretion (IS) and also potentiates the insulin-releasing action of secretagogues such as arginine and sulfonylureas. This potentiating effect is known to be impaired in type 2 diabetic patients, but its cellular mechanisms are unclear. IS and cytosolic Ca(2+) concentration ([Ca(2+)](i)) were measured in mouse islets during perifusion with 3-15 mmol/l glucose (G3-G15, respectively) and pulse or stepwise stimulation with 1-10 mmol/l arginine or 5 250 micromol/l tolbutamide. In G3, arginine induced small increases in [Ca(2+)](i) but no IS. G7 alone only slightly increased [Ca(2+)](i) and IS but markedly potentiated arginine effects on [Ca(2+)](i), which resulted in significant IS (already at 1 mmol/l). For each arginine concentration, both responses further increased at G10 and G15, but the relative change was distinctly larger for IS than [Ca(2+)](i). At all glucose concentrations, tolbutamide dose dependently increased [Ca(2+)](i) and IS with thresholds of 25 micromol/l for [Ca(2+)](i) and 100 micromol/l for IS at G3 and of 5 micromol/l for both at G7 and above. Between G7 and G15, the effect of tolbutamide on [Ca(2+)](i) increased only slightly, whereas that on IS was strongly potentiated. The linear relationship between IS and [Ca(2+)](i) at increasing arginine or tolbutamide concentrations became steeper as the glucose concentration was raised. Thus glucose augmented more the effect of each agent on IS than that on [Ca(2+)](i). In conclusion, glucose potentiation of arginine- or tolbutamide induced IS involves increases in both the rise of [Ca(2+)](i) and the action of Ca(2+) on exocytosis. This dual mechanism must be borne in mind to interpret the alterations of the potentiating action of glucose in type 2 diabetic patients. PMID- 16249258 TI - PilZ domain is part of the bacterial c-di-GMP binding protein. AB - Recent studies identified c-di-GMP as a universal bacterial secondary messenger regulating biofilm formation, motility, production of extracellular polysaccharide and multicellular behavior in diverse bacteria. However, except for cellulose synthase, no protein has been shown to bind c-di-GMP and the targets for c-di-GMP action remain unknown. Here we report identification of the PilZ ("pills") domain (Pfam domain PF07238) in the sequences of bacterial cellulose synthases, alginate biosynthesis protein Alg44, proteins of enterobacterial YcgR and firmicute YpfA families, and other proteins encoded in bacterial genomes and present evidence indicating that this domain is (part of) the long-sought c-di-GMP-binding protein. Association of the PilZ domain with a variety of other domains, including likely components of bacterial multidrug secretion system, could provide clues to multiple functions of the c-di-GMP in bacterial pathogenesis and cell development. PMID- 16249259 TI - Integrating time-course microarray gene expression profiles with cytotoxicity for identification of biomarkers in primary rat hepatocytes exposed to cadmium. AB - MOTIVATION: DNA microarrays can provide information about the expression levels of thousands of genes simultaneously at the transcriptomic level, while conventional cell viability and cytotoxicity measurement methods provide information about the biological functions at the cellular level. Integrating these data at different levels provides a promising approach for evaluating or predicting how cells respond to chemical exposure. It is important to investigate the multi-scale biological system in a systematic way to better understand the gene regulation networks and signal transduction pathways involved in the cellular responses to environmental factors. RESULTS: Primary rat hepatocytes were exposed to cadmium acetate at 0, 1.25 and 2 microM. mRNA expression profiles at 0, 3, 6, 12 and 24 h were measured using the Affymetrix RatTox U34 GeneChip arrays. Simultaneously, cytotoxicity was assessed by lactase dehydrogenase leakage assay. Gene expression profiles at different time points were used to evaluate cytotoxicity at subsequent time points using partial least squares, and it was found that gene expression profiles at 0 h had the best prediction accuracy for the cytotoxicity observed at 12 h. Some biomarkers whose expression profiles showed strong relationship with cytotoxicity were identified and the underlying pathways were reconstructed to illustrate how hepatocytes respond to cadmium exposure. Permutation studies were also applied to assess the reliability of the predictive models. AVAILABILITY: Matlab source code is available upon request and DNA microarray data are available at GEO (http://www.ncbi.nlm.nih.gov/geo). PMID- 16249260 TI - Gene selection using support vector machines with non-convex penalty. AB - MOTIVATION: With the development of DNA microarray technology, scientists can now measure the expression levels of thousands of genes simultaneously in one single experiment. One current difficulty in interpreting microarray data comes from their innate nature of 'high-dimensional low sample size'. Therefore, robust and accurate gene selection methods are required to identify differentially expressed group of genes across different samples, e.g. between cancerous and normal cells. Successful gene selection will help to classify different cancer types, lead to a better understanding of genetic signatures in cancers and improve treatment strategies. Although gene selection and cancer classification are two closely related problems, most existing approaches handle them separately by selecting genes prior to classification. We provide a unified procedure for simultaneous gene selection and cancer classification, achieving high accuracy in both aspects. RESULTS: In this paper we develop a novel type of regularization in support vector machines (SVMs) to identify important genes for cancer classification. A special nonconvex penalty, called the smoothly clipped absolute deviation penalty, is imposed on the hinge loss function in the SVM. By systematically thresholding small estimates to zeros, the new procedure eliminates redundant genes automatically and yields a compact and accurate classifier. A successive quadratic algorithm is proposed to convert the non differentiable and non-convex optimization problem into easily solved linear equation systems. The method is applied to two real datasets and has produced very promising results. AVAILABILITY: MATLAB codes are available upon request from the authors. PMID- 16249261 TI - Automatic term list generation for entity tagging. AB - MOTIVATION: Many entity taggers and information extraction systems make use of lists of terms of entities such as people, places, genes or chemicals. These lists have traditionally been constructed manually. We show that distributional clustering methods which group words based on the contexts that they appear in, including neighboring words and syntactic relations extracted using a shallow parser, can be used to aid in the construction of term lists. RESULTS: Experiments on learning lists of terms and using them as part of a gene tagger on a corpus of abstracts from the scientific literature show that our automatically generated term lists significantly boost the precision of a state-of-the-art CRF based gene tagger to a degree that is competitive with using hand curated lists and boosts recall to a degree that surpasses that of the hand-curated lists. Our results also show that these distributional clustering methods do not generate lists as helpful as those generated by supervised techniques, but that they can be used to complement supervised techniques so as to obtain better performance. AVAILABILITY: The code used in this paper is available from http://www.cis.upenn.edu/datamining/software_dist/autoterm/ PMID- 16249262 TI - Haplotype-based linkage disequilibrium mapping via direct data mining. AB - MOTIVATION: With the availability of large-scale, high-density single-nucleotide polymorphism markers and information on haplotype structures and frequencies, a great challenge is how to take advantage of haplotype information in the association mapping of complex diseases in case-control studies. RESULTS: We present a novel approach for association mapping based on directly mining haplotypes (i.e. phased genotype pairs) produced from case-control data or case parent data via a density-based clustering algorithm, which can be applied to whole-genome screens as well as candidate-gene studies in small genomic regions. The method directly explores the sharing of haplotype segments in affected individuals that are rarely present in normal individuals. The measure of sharing between two haplotypes is defined by a new similarity metric that combines the length of the shared segments and the number of common alleles around any marker position of the haplotypes, which is robust against recent mutations/genotype errors and recombination events. The effectiveness of the approach is demonstrated by using both simulated datasets and real datasets. The results show that the algorithm is accurate for different population models and for different disease models, even for genes with small effects, and it outperforms some recently developed methods. PMID- 16249263 TI - Integrated minimum-set primers and unique probe design algorithms for differential detection on symptom-related pathogens. AB - MOTIVATION: Differential detection on symptom-related pathogens (SRP) is critical for fast identification and accurate control against epidemic diseases. Conventional polymerase chain reaction (PCR) requires a large number of unique primers to amplify selected SRP target sequences. With multiple-use primers (mu primers), multiple targets can be amplified and detected in one PCR experiment under standard reaction condition and reduced detection complexity. However, the time complexity of designing mu-primers with the best heuristic method available is too vast. We have formulated minimum-set mu-primer design problem as a set covering problem (SCP), and used modified compact genetic algorithm (MCGA) to solve this problem optimally and efficiently. We have also proposed new strategies of primer/probe design algorithm (PDA) on combining both minimum-set (MS) mu-primers and unique (UniQ) probes. Designed primer/probe set by PDA MS/UniQ can amplify multiple genes simultaneously upon physical presence with minimum-set mu-primer amplification (MMA) before intended differential detection with probes-array hybridization (PAH) on the selected target set of SRP. RESULTS: The proposed PDA-MS/UniQ method pursues a much smaller number of primers set compared with conventional PCR. In the simulation experiment for amplifying 12 669 target sequences, the performance of our method with 68% reduction on required mu-primers number seems to be superior to the compared heuristic approaches in both computation efficiency and reduction percentage. Our integrated PDA-MS/UniQ method is applied to the differential detection on 9 plant viruses from 4 genera with MMA and PAH of 11 mu-primers instead of 18 unique ones in conventional PCR while amplifying overall 9 target sequences. The results of wet lab experiments with integrated MMA-PAH system have successfully validated the specificity and sensitivity of the primers/probes designed with our integrated PDA-MS/UniQ method. PMID- 16249264 TI - ISER: selection of differentially expressed genes from DNA array data by non linear data transformations and local fitting. AB - SUMMARY: This report describes an algorithm (intensity-dependent selection of expression ratios or ISER) developed to analyse DNA array data by optimizing the selection of genes with the most significant variations in expression amongst two RNA samples. The algorithm is designed for use when little or no replication of array hybridizations is available. PMID- 16249265 TI - GSMA: software implementation of the genome search meta-analysis method. AB - Meta-analysis can be used to pool results of genome-wide linkage scans. This is of great value in complex diseases, where replication of linked regions occurs infrequently. The genome search meta-analysis (GSMA) method is widely used for this analysis, and a computer program is now available to implement the GSMA. PMID- 16249266 TI - Large-scale prokaryotic gene prediction and comparison to genome annotation. AB - MOTIVATION: Prokaryotic genomes are sequenced and annotated at an increasing rate. The methods of annotation vary between sequencing groups. It makes genome comparison difficult and may lead to propagation of errors when questionable assignments are adapted from one genome to another. Genome comparison either on a large or small scale would be facilitated by using a single standard for annotation, which incorporates a transparency of why an open reading frame (ORF) is considered to be a gene. RESULTS: A total of 143 prokaryotic genomes were scored with an updated version of the prokaryotic genefinder EasyGene. Comparison of the GenBank and RefSeq annotations with the EasyGene predictions reveals that in some genomes up to approximately 60% of the genes may have been annotated with a wrong start codon, especially in the GC-rich genomes. The fractional difference between annotated and predicted confirms that too many short genes are annotated in numerous organisms. Furthermore, genes might be missing in the annotation of some of the genomes. We predict 41 of 143 genomes to be over-annotated by >5%, meaning that too many ORFs are annotated as genes. We also predict that 12 of 143 genomes are under-annotated. These results are based on the difference between the number of annotated genes not found by EasyGene and the number of predicted genes that are not annotated in GenBank. We argue that the average performance of our standardized and fully automated method is slightly better than the annotation. PMID- 16249267 TI - Medical record review of deaths, unexpected intensive care unit admissions, and clinician referrals: detection of adverse events and insight into the system. AB - AIMS AND METHODS: To determine whether a programme of continuous medical record review of deaths, unexpected intensive care unit (ICU) admissions, and admissions referred by medical and nursing staff for specific review, would provide a range of adverse events from which to gain insight into the healthcare system of a large paediatric referral hospital. A quality assurance programme was commenced in 1996. RESULTS: Over a six year period there were 103 255 admissions; 1612 (1.6%) records were reviewed, from which 325 adverse events were detected. Events were associated with operations, procedures and anaesthesia (56.5%), diagnosis and therapy (24%), drug and fluid management (12.6%), and system issues (7%). Medical records were reviewed from 23 of the 28 clinical units. Review of the records and analysis of the adverse events triggered many system changes. CONCLUSIONS: The findings suggest that continuous medical record review may be a valuable method for the detection of adverse events and identifying system issues in children's hospitals. PMID- 16249268 TI - Scombrotoxic fish poisoning. PMID- 16249269 TI - Rituximab treatment of refractory rheumatoid arthritis. AB - OBJECTIVE: To review published literature using rituximab for treatment of refractory rheumatoid arthritis (RA). DATA SOURCES: An English-language literature search was conducted using MEDLINE (1966-May 2005) and EMBASE (1980 May 2005). References of identified articles were subsequently reviewed for additional data. DATA SYNTHESIS: Evidence suggests that B lymphocyte depletion in patients suffering from refractory RA may be a key component in the interruption of the disease pathogenesis. Successful depletion of B lymphocytes with rituximab in patients with RA has been reported in case reports, open-label pilot studies, and a randomized, double-blind, placebo-controlled trial. CONCLUSIONS: Based on the limited published data, rituximab, when used in combination with other agents (ie, cyclophosphamide or methotrexate), appears to be a reasonable treatment option for refractory RA. However, additional controlled trials need to be conducted to further define optimal dosing, response rates, comparative long-term efficacy, and RA treatment algorithm placement of rituximab in this patient population. PMID- 16249270 TI - Efaproxiral: a radiation enhancer used in brain metastases from breast cancer. AB - OBJECTIVE: To review the mechanism of action and clinical data of efaproxiral use in brain metastases of breast cancer. DATA SOURCES: Articles were identified through MEDLINE (1966-June 2005) and EMBASE (1980-May 2005) searches using the key words efaproxiral and RSR13. Published abstracts over the previous 10 years from various scientific meetings, including American Society of Clinical Oncology and San Antonio Breast Cancer Symposium, were also searched for investigations of efaproxiral. Data on efaproxiral were also provided by Allos Therapeutics. STUDY SELECTION AND DATA EXTRACTION: All published clinical data in humans regarding efaproxiral use in brain metastases from breast cancer were selected for this review. In addition, published studies in humans that discussed the pharmacokinetics, pharmacodynamics, and safety of efaproxiral were evaluated. DATA SYNTHESIS: Efaproxiral is a synthetic allosteric modifier of hemoglobin that results in a shift of the hemoglobin oxygen dissociation curve to the right. Therefore, oxygen is more readily released from hemoglobin into tissues. Efaproxiral demonstrated a significant survival benefit when used as a radiation enhancer in patients with brain metastases originating from breast cancer. The safety profile of efaproxiral and improved survival rates make this agent advantageous over radiation alone. Further investigation and results from the ongoing clinical trials will help to define the role of efaproxiral in clinical practice. CONCLUSIONS: Efaproxiral is the first synthetic allosteric modifier to demonstrate significant improvement in survival in patients undergoing radiation therapy for brain metastases of breast cancer. Validation of this effect in ongoing clinical trials will be important in determining the role of efaproxiral in brain metastases from breast cancer. PMID- 16249271 TI - Effect of vitamin A on fracture risk. AB - OBJECTIVE: To describe current data evaluating the effect of vitamin A intake on fracture risk. DATA SOURCES: A literature search using MEDLINE (1966-March 2005) was conducted using the search terms bone density, fractures, osteoporosis, retinol, and vitamin A to identify published studies evaluating the effects of vitamin A on bone. DATA SYNTHESIS: Studies evaluating vitamin A consumption and fracture risk were reviewed. Current data suggest a potential inverse relationship between excess vitamin A consumption and bone mineral density leading to an increased risk for fracture. CONCLUSIONS: Although current data are limited, consumption of large amounts of vitamin A may be associated with decreased bone mineral density and increased fracture risk. Until further information is available, patients should be made aware of the potential risks of consuming vitamin A in amounts that exceed the recommended dietary allowance. Further research is needed to clarify the relationship between vitamin A and fracture risk. PMID- 16249273 TI - Angiotensin II stimulation of VEGF mRNA translation requires production of reactive oxygen species. AB - ANG II, a mediator of renal injury in diabetic renal disease, promotes vascular endothelial growth factor (VEGF) mRNA translation in proximal tubular epithelial (MCT) cells (Feliers D, Duraisamy S, Barnes JL, Ghosh-Choudhury G, and Kasimath BS. Am J Physiol Renal Physiol 288: F521-F529, 2005). The mechanism by which ANG II elicits this effect is not known. ANG II is known to induce oxidative stress and the rapidity of the effect suggested a role for reactive oxygen species (ROS). The aim of this study is to test the hypothesis that ANG II regulates VEGF mRNA translation in MCT cells through ROS production. In MCT cells exposed to 1 nM ANG II, ROS production was increased in a time-dependent manner. Inhibition of ROS production by N-acetylcysteine (NAC), a precursor of glutathione, and diphenyleneiodonium (DPI), an inhibitor of flavoproteins that include NAD(P)H oxidase, prevented ANG II-stimulated VEGF protein expression. NAC and DPI also inhibited phosphorylation of 4E-BP1 on Thr46 and association of eIF4E with eIF4G, steps that are important in the initiation phase of mRNA translation. NAC and DPI also blocked Akt activation which is required for 4E-BP1 phosphorylation. LY 294002, a selective phosphatidylinositol (PI 3-kinase) inhibitor, did not prevent ROS accumulation in response to ANG II, whereas DPI blocked ANG II activation of PI 3-kinase, demonstrating that ROS production is upstream of the PI 3-kinase signaling pathway. Preincubation with catalase abolished ANG II stimulation of VEGF expression and mRNA translation, suggesting involvement of hydrogen peroxide (H(2)O(2)). H(2)O(2) reproduced the effects of ANG II on VEGF expression and aforementioned parameters of mRNA translation. Finally, neither preincubation of MCT cells with specific inhibitors of the mitochondrial respiratory chain nor inactivation of the mitochondrial respiratory chain in MCT cells prevented ANG II stimulation of VEGF expression. Inhibition of nitric oxide synthase by l-NAME had no effect on ANG II stimulation of VEGF expression. These data show that ROS, generated probably through activation of an NAD(P)H oxidase, mediate ANG II stimulation of VEGF mRNA translation. PMID- 16249272 TI - Longitudinal study of urinary excretion of phosphate, calcium, and uric acid in mutant NHERF-1 null mice. AB - NHERF-1 binds numerous renal protein targets, including the proximal tubule transporters Na(+)/H(+) exchanger 3 (NHE3) and Na(+)-phosphate cotransporter 2a (Npt2a). Young NHERF-1(-/-) male mice display defective targeting of Npt2a to apical membranes in the renal proximal tubule and manifest hypophosphatemia and increased urinary excretion of phosphate. The present studies describe the changes in the urinary excretion of phosphate, calcium, uric acid, and sodium in male and female wild-type and NHERF-1 null mice over a time period from 12 to 54 wk of age. Young male and female NHERF-1(-/-) mice demonstrated increased urinary excretion of phosphate and urine phosphate/creatinine ratios. There was an age related decline in the phosphate/creatinine ratio in mutant mice such that there were no differences between wild-type and NHERF-1(-/-) by 24 to 30 wk of age despite the continued presence of hypophosphatemia. Male and female NHERF-1 null mice also demonstrate increased urine calcium/creatinine and uric acid/creatinine ratios compared with wild-type controls. These studies indicate defects in the renal tubule transport of phosphate, calcium, and uric acid in NHERF-1(-/-) male and female mice that could account for the increased deposition of calcium in the papilla of null mice. PMID- 16249274 TI - Functional polymorphisms in the alpha-subunit of the human epithelial Na+ channel increase activity. AB - Activity of the epithelial Na(+) channel (ENaC) is limiting for Na(+) reabsorption at the distal nephron. Gain-of-function mutations in ENaC cause Liddle's syndrome: a severe form of inheritable hypertension. Several polymorphisms in alpha-hENaC possibly associated with abnormal Na(+) handling by the kidney and the salt-sensitive hypertension prevalent in black populations have been reported. The functional effects of alpha-hENaC polymorphisms on channel activity, however, remain controversial and have not been directly tested in a mammalian background. We ask here whether polymorphisms at positions 334, 618, and 663 in alpha-hENaC influence channel activity. Activity of wild-type (A334, C618, A663) and polymorphic ENaC expressed in Chinese hamster ovary cells was assessed with patch-clamp electrophysiology. While the A334T polymorphism had little effect on macroscopic ENaC currents, the C618F and A663T polymorphisms significantly increased ENaC activity >3.3- and 1.6-fold, respectively. Similarly, polymorphic ENaC had greater activity compared with wild-type channels in excised patches with activity of C618F and A663T channels increased 3.8- and 2.6-fold, respectively. Unitary channel conductances and reversal potentials were not different for polymorphic and wild-type ENaC. Increases in activity resulted primarily from increases in the apparent number of active (polymorphic) channels in the plasma membrane. Moreover, addition of a reducing agent to the cytosol significantly increased activity of wild-type ENaC equal to that of C618F polymorphic channels but had no effect on these latter channels. These results are consistent with the C618F and A663T polymorphisms leading to elevated ENaC activity with the possibility that they facilitate altered Na(+) handling by the kidney. PMID- 16249275 TI - Osmolarity-induced renin secretion from kidneys: evidence for readily releasable renin pools. AB - Our study aimed to characterize the influence of changes in extracellular osmolarity on renin secretion from the whole kidney. For this purpose, the osmolarity of the perfusion medium of isolated rat or mouse kidneys was either decreased by lowering the NaCl concentration by 20% or was increased up to 133% by the addition of various salts or sugars. It turned out that changes in osmolarity led to instantaneous transient changes followed by a plateau of renin secretion, in that increases in osmolarity stimulated renin secretion, whereas decreases attenuated renin secretion. The peak amplitude of changes in renin secretion was related to steady-state renin secretion rates before the osmotic challenge but was independent of the maneuver used to modulate steady-state renin secretion. Osmolarity-induced changes in renin secretion were more related to relative rather than to absolute changes in osmolarity and were not dependent on the formation of nitric oxide or of prostanoids and did not require Na-K-2Cl cotransport function or swelling-activated chloride channels. Moreover, we obtained evidence that the pool of renin secretion excitable by hyperosmolarity is exhaustible and that its complete refilling takes at least 2 min. The observed behavior of renin secretion fits the concept about exocytosis proposing the existence of different pools of committed secretory vesicles, which have not yet undergone the final modification for initiation of exocytosis. Probably, a pool of readily releasable vesicles determines steady-state secretion rates from kidneys. PMID- 16249276 TI - Tubular mitochondrial alterations in neonatal rats subjected to RAS inhibition. AB - Pharmacological interruption of the angiotensin II (ANG II) type 1 receptor signaling during nephrogenesis in rats perturbs renal tubular development. This study aimed to further investigate tubular developmental defects in neonatal rats subjected to ANG II inhibition with enalapril. We evaluated tubular ultrastructural changes using electron microscopy and estimated spectrophotometrically activity or concentrations of succinate dehydrogenase (SDH), cytochromes a and c, which are components of mitochondrial respiratory chain, on postnatal days 2 and 9 (PD2 and PD9). Renal expression of sodium potassium adenosinetriphosphatase (Na(+)-K(+)-ATPase) and two reflectors of mitochondrial biogenesis [mitochondrial transcription factor A (TFAM) and translocase of outer mitochondrial membrane 20 (TOM20)] also were studied using Western immunoblotting and immunohistochemistry. Enalapril disrupted inner mitochondrial membranes of developing cortical and medullary tubular cells on PD2 and PD9. These findings were paralleled by impaired mitochondrial respiratory function, as revealed from the changes in components of the mitochondrial respiratory chain, such as decreased cytochrome c level in the cortex and medulla on PD2 and PD9, decreased cytochrome a level in the cortex and medulla on PD2, and diminished cortical SDH activity on PD2 and PD9. Moreover, tubular expression of the most active energy-consuming pump Na(+)-K(+)-ATPase was decreased by enalapril treatment. Renal expression of TFAM and TOM20 was not altered by neonatal enalapril treatment. Because nephrogenesis is a highly energy-demanding biological process, with the energy being utilized for renal growth and transport activities, the structural-functional alterations of the mitochondria induced by neonatal enalapril treatment may provide the propensity for the tubular developmental defect. PMID- 16249277 TI - Aberrant expression of human luteinizing hormone receptor by adrenocortical cells is sufficient to provoke both hyperplasia and Cushing's syndrome features. AB - CONTEXT: Aberrant expression of LH/human chorionic gonadotropin (hCG) receptor has been suggested in several cases of bilateral macronodular adrenal hyperplasia with Cushing's syndrome. The cortisol production is then directly controlled by endogenous secretion of LH/hCG. However, the direct involvement of this aberrant LH/hCG receptor expression in the development of the hyperplasia has not been demonstrated. Moreover in most cases, whenever investigated, the aberrant expression of LH/hCG receptor has been associated with the ectopic expression of other G protein-coupled receptors such as gastric inhibitory polypeptide, serotonin, or vasopressin receptors. OBJECTIVE: The aim of this study was to explore the action of LH/hCG receptor on the development of adrenal hyperplasia. RESULTS: The ectopic expression of this single nonmutated gene transduced into bovine adrenocortical cells was sufficient to induce not only the aberrant cortisol secretion but also hyperproliferation and benign transformation. The cells were transplanted beneath the kidney capsule of adrenalectomized immunodeficient mice. Only the cells expressing the LH/hCG receptor gene formed an enlarged tissue with a high proliferation rate. The tissue expressing LH/hCG receptor was responsible for elevated plasma cortisol and decreased plasma ACTH levels in transplanted mice. These animals displayed physiological changes similar to those of patients with Cushing's syndrome, including muscle atrophy, thin skin, spleen atrophy, and hyperglycemia. CONCLUSIONS: These results demonstrate that a single genetic event such as the inappropriate expression of the nonmutated LH/hCG receptor gene is sufficient to initiate the phenotypic changes that cause the development of a benign adrenocortical tumor. PMID- 16249278 TI - High frequency of loss of heterozygosity in imprinted, compared with nonimprinted, genomic regions in follicular thyroid carcinomas and atypical adenomas. AB - CONTEXT: Many mammalian genes that are imprinted regulate cell growth, differentiation, and apoptosis. Because imprinting silences one of the two alleles, resulting in functional haploinsufficiency, we hypothesized that loss of heterozygosity (LOH) at an imprinted locus may result in the deletion of the only functional copy of an imprinted tumor suppressor gene. OBJECTIVE: The goal of this study was to specifically address this hypothesis that in thyroid neoplasias loss of imprinted loci becomes enriched during oncogenesis. DESIGN: In total, thyroid tissue was obtained from 72 patients with thyroid neoplasias comprising 34 follicular thyroid carcinomas (FTCs) and 38 follicular adenomas. We performed PCR-based LOH analysis of DNA from paired normal-tumor samples using 18 markers mapped to imprinted regions (IR) and 13 markers in nonimprinted regions (NIR). RESULTS: Overall LOH frequencies for the IR markers were 26% for the adenomas and 38% for the carcinomas. In the NIR, the overall LOH frequency was 23 and 26% for adenomas and FTCs, respectively. The difference in LOH frequencies between IR and NIR was statistically significant only for the carcinomas (P = 0.001), although there was a similar trend for the atypical adenomas (ATY, P = 0.06). CONCLUSIONS: Our observations suggest that IRs are more prone to genomic instability in FTCs. The fact that the ATY trended toward differential IR/NIR LOH, similar to FTC, may suggest that loss of IR might be instrumental in the adenoma-carcinoma sequence in thyroid carcinogenesis and that ATY could be an important intermediate in this pathway. PMID- 16249279 TI - Reactive oxygen species-induced oxidative stress in the development of insulin resistance and hyperandrogenism in polycystic ovary syndrome. AB - CONTEXT: Insulin resistance and chronic low level inflammation are often present in women with polycystic ovary syndrome (PCOS). OBJECTIVE: The purpose of this study was to determine the effects of hyperglycemia on reactive oxygen species (ROS) generation from mononuclear cells (MNCs) in PCOS. DESIGN: This was a prospective controlled study. SETTING: The study was conducted at an academic medical center. PATIENTS: The study population consisted of 16 women with PCOS (eight lean, eight obese) and 15 age- and body composition-matched controls (eight lean, seven obese). MAIN OUTCOME MEASURES: Insulin sensitivity was derived from a 2-h, 75-g oral glucose tolerance test (IS(OGTT)). ROS generation and p47(phox) protein expression were quantitated from MNCs obtained from blood drawn fasting and 2 h after glucose ingestion. RESULTS: IS(OGTT) was lower in PCOS, compared with controls (3.1 +/- 0.3 vs. 6.3 +/- 0.9, P < 0.003). The percent change in ROS generation from MNCs was higher in lean and obese PCOS, compared with lean controls (138.8 +/- 21.3 and 154.2 +/- 49.1 vs. 0.6 +/- 12.7, P < 0.003). The percent change in ROS generation from MNCs correlated positively with glucose area under the curve (r = 0.38, P < 0.05), and plasma levels of testosterone (r = 0.59, P < 0.002) and androstenedione (r = 0.50, P < 0.009). The percent change in p47(phox) from MNCs was also higher in lean and obese PCOS, compared with lean controls (36.2 +/- 18.2 and 39.1 +/- 8.0 vs. -13.7 +/- 8.7, P < 0.02), and correlated negatively with IS(OGTT) (r = -0.39, P < 0.05). CONCLUSION: ROS generation from MNCs in response to hyperglycemia is increased in PCOS independent of obesity. The resultant oxidative stress may contribute to a proinflammatory state that induces insulin resistance and hyperandrogenism in women with this disorder. PMID- 16249281 TI - Cross-sectional associations of resistin, coronary heart disease, and insulin resistance. AB - CONTEXT: Recently, resistin was found to be present in atherosclerotic lesions in apoE(-/-) mice. Resistin may be associated with inflammation and atherosclerosis in humans; however, the role of resistin in human disease remains controversial. OBJECTIVE: This study assesses cross-sectional relationships of resistin with coronary heart disease (CHD). DESIGN, SETTING, AND PARTICIPANTS: Blood samples from the third examination of the Strong Heart Study (SHS)--the largest study of CHD in American Indians--were used. Cases who had suffered previous myocardial infarction (n = 100) were selected randomly from the three SHS sites and matched for study site and sex with controls who had no history of cardiovascular disease (CHD or stroke) (n = 100). MAIN OUTCOME MEASURE: Resistin levels by enzyme-linked immunosorbent assay method in cases and controls was the main outcome measure. RESULTS: Resistin levels were higher in cases than controls [median (interquartile range): 3.4 (2.5-4.7) vs. 2.8 (2.1-4.0) ng/ml; P = 0.003] and had univariate correlations with age (Spearman r = 0.21; P < 0.002), fasting insulin (r = 0.21; P = 0.003), insulin resistance by homeostasis model (r = 0.22; P = 0.04), albumin to creatinine ratio (r = 0.19; P = 0.01), and fibrinogen (r = 0.34; P < 0.0001). Cases were more likely to have diabetes (cases 67%; controls 41%; P < 0.0001) but had similar body mass index (cases 31.4 +/- 5.4; controls 30.7 +/- 6.3; P = 0.85). Resistin levels were higher in participants with established nephropathy (albumin to creatinine ratio >300 mg/g, n = 26) compared with those with normo- (n = 122) or microalbuminuria (n = 42). In multivariate analysis, nephropathy (P = 0.0013) but not previous myocardial infarction (P = 0.12) was significantly associated with resistin. CONCLUSIONS: Resistin is not independently associated with CHD. Resistin is elevated in survivors of myocardial infarction; however, this reflects a novel association of raised resistin with diabetic nephropathy. PMID- 16249280 TI - Adolescent girls with polycystic ovary syndrome have an increased risk of the metabolic syndrome associated with increasing androgen levels independent of obesity and insulin resistance. AB - CONTEXT: Adult women with polycystic ovary syndrome (PCOS) have an increased prevalence of the metabolic syndrome (MBS). The prevalence of MBS is also increasing in adolescents. OBJECTIVE: Our objective was to test the hypothesis that the prevalence of MBS is increased in adolescent girls with PCOS compared with the general population and to determine the factors associated with an increased risk of the MBS in PCOS. DESIGN AND SETTING: We conducted a cross sectional case-control study at academic medical centers with general clinical research centers. PARTICIPANTS: Participants included 49 adolescent girls with PCOS and 165 girls from the Third National Health and Nutrition Examination Survey (NHANES III) adolescent population of similar age and ethnic background. MAIN OUTCOME MEASURE: We assessed the prevalence of MBS according to currently proposed adolescent MBS criteria. RESULTS: Thirty-seven percent of adolescent girls with PCOS had MBS compared with 5% of NHANES III girls (P < 0.0001). None of the girls of normal body mass index (BMI) had MBS, whereas 11% of overweight and 63% of obese girls with PCOS had MBS compared with 0 and 32% of NHANES III girls, respectively. Girls with PCOS were 4.5 times more likely to have MBS than age-matched NHANES III girls after adjusting for BMI (odds ratio, 4.5; 95% confidence interval, 1.1-17.7; P = 0.03). The odds of having the MBS were 3.8 times higher for every quartile increase in bioavailable testosterone in girls with PCOS after adjusting for BMI and insulin resistance (odds ratio, 3.8; 95% confidence interval, 1.4-10.2; P = 0.008). CONCLUSIONS: Adolescent girls with PCOS have a higher prevalence of MBS than the general adolescent population. Hyperandrogenemia is a risk factor for MBS independent of obesity and insulin resistance. PMID- 16249282 TI - Differential effects of sex hormones on peri- and endocortical bone surfaces in pubertal girls. AB - CONTEXT: The role of sex steroids in bone growth in pubertal girls is not yet clear. Bone biomarkers are indicators of bone metabolic activity, but their value in predicting bone quality has not been studied in growing girls. OBJECTIVE: This study examines the association of sex hormones and bone markers with bone geometry and density in pubertal girls. DESIGN: The study was designed as a 2-yr longitudinal study in pubertal girls. Measurements were performed at baseline and at 1- and 2-yr follow-ups. SETTING: The study was conducted in a university laboratory. PARTICIPANTS: A total of 258 10- to 13-yr-old healthy girls at the baseline participated. METHODS: Peripheral quantitative computed tomography was used to scan the left tibial shaft. Serum 17beta-estradiol (E2), testosterone (T), SHBG, osteocalcin (OC), bone-specific alkaline phosphatase, and tartrate resistant acid phosphatase isoform 5b were assessed. Data were analyzed using hierarchical linear models with random effect. RESULTS: E2 was a positive predictor for total bone mineral density (BMD), cortical thickness, and a negative predictor for endocortical circumference but had no predictive value for total bone cross-sectional area or periosteal circumference. T was a positive predictor for total cross-sectional area and periosteal circumference as well as endocortical circumference, and a negative predictor for total BMD. OC was negatively correlated with cortical BMD (R2 = 0.325; P < 0.001). CONCLUSIONS: In pubertal girls, E2 and T have different influences on bone properties at the long bone shaft. The results suggest that, at the endocortical surface, E2 inhibits bone resorption during rapid growth, and later, after menarche, acts at higher concentrations to promote bone formation. At the periosteal surface, T promotes bone formation, whereas E2 does not affect it. In addition, OC might be used as a predictor of cortical BMD. PMID- 16249283 TI - Patients with multiple endocrine neoplasia type 1 with gastrinomas have an increased risk of severe esophageal disease including stricture and the premalignant condition, Barrett's esophagus. AB - CONTEXT: Multiple endocrine neoplasia type 1 (MEN1) patients frequently develop Zollinger-Ellison syndrome (MEN1/ZES). Although esophageal reflux symptoms are common in these patients, little is known about long-term occurrence of severe peptic esophageal disease including strictures and Barrett's esophagus (BE). OBJECTIVE: The objective of the study was to prospectively analyze the frequency of severe peptic esophageal disease in ZES patients with and without MEN1. SETTING: The study was conducted at a tertiary care research center. PATIENTS: Two hundred ninety-five patients (80 = MEN1/ZES, 215 = sporadic ZES) participated in a prospective study. INTERVENTIONS AND OUTCOME MEASURES: Assessment of MEN1, acid hypersecretion, upper gastrointestinal endoscopy/biopsies, and tumor status were measured initially and at each follow-up. Esophageal manometry was performed in 89 patients. Frequency and type of esophageal disease were correlated with clinical/laboratory/tumoral features of ZES/MEN1. RESULTS: In MEN1/ZES patients, esophageal stricture was 3-fold higher, BE 5-fold higher, and dysplasia 8-fold higher, and one patient died of esophageal adenocarcinoma. Esophageal symptoms were more frequent or severe in MEN1/ZES, but known risk factors for severe esophageal disease and ZES-specific features did not differ between MEN1/ZES and sporadic ZES. In MEN1/ZES, the onset of ZES was 10 yr earlier, and H2-antagonists were used longer and at lower doses. MEN1/ZES patients with esophageal disease differed from those without in that ZES diagnosis was delayed longer, esophageal symptoms were more frequent or severe, hiatal hernias were more frequent, esophagitis or pyloric scarring was more common, basal acid output was higher, and hyperparathyroidism was underdiagnosed. CONCLUSIONS: This study shows that MEN1/ZES patients have a higher incidence of severe esophageal disease including the premalignant condition BE and identifies factors important for their pathogenesis that need to be incorporated into their long-term treatment. PMID- 16249284 TI - Prevalence and predictors of the metabolic syndrome in women with polycystic ovary syndrome. AB - CONTEXT: Polycystic ovary syndrome (PCOS) and the metabolic syndrome have many features in common and may share the same pathogenesis. OBJECTIVE: This study was performed to determine the prevalence and predictors of the metabolic syndrome in PCOS. DESIGN: The clinical, hormonal, and oral glucose tolerance test results were analyzed in 394 PCOS women who were screened for participation in a multicenter trial to evaluate the effects of troglitazone on ovulation and hirsutism. SETTING: A multicenter clinical trial is presented. PATIENTS OR OTHER PARTICIPANTS: The subjects were women with PCOS who had or lacked the metabolic syndrome. MAIN OUTCOME MEASURES: Waist circumference, fasting glucose, high density lipoprotein cholesterol and triglyceride concentrations, and blood pressure were the main outcome measures. RESULTS: Twenty-six (6.6%) subjects had diabetes; among the 368 nondiabetics, the prevalence for individual components comprising the metabolic syndrome were: waist circumference greater than 88 cm in 80%, high-density lipoprotein cholesterol less than 50 mg/dl in 66%, triglycerides greater than or equal to 150 mg/dl in 32%, blood pressure greater than or equal to 130/85 mm Hg in 21%, and fasting glucose concentrations greater than or equal to 110 mg/dl in 5%. Three or more of these individual criteria were present in 123 (33.4%) subjects overall. The prevalence of the metabolic syndrome did not differ significantly between racial/ethnic groups. The prevalence of the metabolic syndrome from lowest to highest quartile of free testosterone concentration was 19.8, 31.3, 46.9, and 35.0%, respectively [P = 0.056 adjusted for body mass index (BMI)]. None of the 52 women with a BMI less than 27.0 kg/m2 had the metabolic syndrome; those in the top BMI quartile were 13.7 times more likely (95% confidence interval, 5.7-33.0) to have the metabolic syndrome compared with those in the lowest quartile. Thirty-eight percent of those with the metabolic syndrome had impaired glucose tolerance compared with 19% without the metabolic syndrome (P < 0.001). CONCLUSIONS: The metabolic syndrome and its individual components are common in PCOS, particularly among women with the highest insulin levels and BMI. Hyperinsulinemia is a likely common pathogenetic factor for both PCOS and the metabolic syndrome. PMID- 16249285 TI - Ethnicity modifies the effect of obesity on insulin resistance in pregnancy: a comparison of Asian, South Asian, and Caucasian women. AB - CONTEXT: Women of Asian and South Asian descent are at increased risk of developing gestational diabetes mellitus compared with Caucasians, despite lower body mass index (BMI). Nevertheless, there has been limited study of insulin action during pregnancy in these ethnic groups. OBJECTIVE: The objective of the study was to compare insulin sensitivity in pregnancy in Asian, South Asian, and Caucasian subjects and to determine whether the impact of obesity on insulin action is modified by ethnicity. DESIGN AND PARTICIPANTS: A cross-sectional study was performed in outpatients undergoing oral glucose tolerance testing in late pregnancy. Participants were stratified into three groups: 1) Caucasian (n = 116); 2) South Asian (n = 31); and 3) Asian (n = 28). MAIN OUTCOME MEASURE: Insulin sensitivity was measured using the oral glucose tolerance test (IS(OGTT)) index of M. Matsuda and R. DeFronzo, previously validated in pregnancy. RESULTS: There were no significant ethnic differences in insulin sensitivity despite variation in prepregnancy BMI (Caucasians, 25.2 kg/m(2); South Asians, 23.3 kg/m(2); Asians, 21.4 kg/m(2); overall P = 0.0001). On multiple linear regression analysis, the strongest independent determinants of IS(OGTT) were gestational diabetes mellitus (t = -5.71; P < 0.0001) and BMI (t = -5.43; P < 0.0001). Importantly, both Asian (t = -2.87; P = 0.0047) and South Asian (t = -2.46; P = 0.015) ethnicity also emerged as negative, independent determinants of IS(OGTT). Furthermore, Asian ethnicity significantly modified the association of prepregnancy BMI with IS(OGTT) (interaction term, t = -2.29; P = 0.0231). CONCLUSIONS: Asian and South Asian ethnicity are both independently associated with increased insulin resistance in late pregnancy. Prepregnancy BMI has a much greater effect on insulin resistance in pregnancy in Asian women than in Caucasians. Ethnicity thus emerges as a factor that modulates the effect of obesity on insulin resistance in pregnancy. PMID- 16249286 TI - The long-term predictive accuracy of the short synacthen (corticotropin) stimulation test for assessment of the hypothalamic-pituitary-adrenal axis. AB - CONTEXT: The high-dose short Synacthen (corticotropin) test (SST) is widely used to investigate suspected secondary adrenal insufficiency, but concern remains about falsely reassuring results. OBJECTIVE: Our objective was to evaluate the long-term safety of the SST. METHOD: We retrospectively evaluated the clinical outcome in 178 patients who achieved 30-min cortisol values in the lowest 15th percentile of normal healthy responses. Thirty patients were later excluded because of missing case notes (20 patients) or unsubstantiated pituitary pathology (10 patients). The remaining 148 patients were divided into two groups: group 1, patients with cortisol response between the 5th and 15th percentiles of normal response (551-635 nmol/liter, 98 patients); and group 2, patients with borderline response between the 2.5th and 5th percentiles (510-550 nmol/liter, 50 patients). Patients did not receive routine glucocorticoid therapy, but those in group 2 were advised to take hydrocortisone in case of intercurrent illness. RESULTS: The median follow-up period from the initial SST was 4.2 yr (range, 4 months to 7 yr). A total of 137 patients showed no clinical or biochemical evidence of adrenal insufficiency during follow-up. Of the remaining 11 patients, seven became hypoadrenal after subsequent pituitary surgery or radiotherapy, one patient in group 1 developed adrenal insufficiency at 2 yr, and one patient in group 2 developed adrenal insufficiency at 6 months. The other two patients who were in group 2 had clinical diagnostic uncertainty. CONCLUSION: The high-dose SST is safe for the purpose of excluding clinically significant secondary adrenal insufficiency and is indicated as the first line of investigation for this purpose. PMID- 16249287 TI - Immune phenotype and serum leptin in children with obesity-related liver disease. AB - CONTEXT: Little is known about pathogenesis of obesity-related liver disease in childhood. Data on the relationship among leptin, immunological parameters, and liver disease in obese children are lacking. OBJECTIVE: Thus, the objective of this study was to evaluate immune phenotype and leptin serum levels in obese children with and without obesity-related liver disease. DESIGN: The study was performed in two groups of consecutive obese children: the first formed by children with obesity-related liver disease, diagnosed in the presence of chronic hypertransaminasemia, liver steatosis at ultrasound, and absence of known etiologies; the second composed of children with isolated obesity. In all patients serum leptin, immunoglobulins, peripheral T, B, and natural killer (NK) cells were evaluated. RESULTS: Twenty-three children in the first group and 16 children in the second were considered eligible. Serum leptin was increased in both groups but without any significant difference. No significant correlation was found between leptin and aminotransferases, lipid serum levels, and all tested lymphocyte subpopulations. Patients with obesity-related liver disease showed significantly higher peripheral NK and B cell counts and IgA levels than children with isolated obesity. Furthermore, no correlation was found between severity of liver disease and lymphocyte subpopulations. CONCLUSION: In our study, leptin did not correlate with hepatic steatosis, aminotransferases, and serum lipids. Children with obesity-related liver disease showed significantly higher peripheral NK and B cells and IgA levels. Additional studies are required to define the pathogenetic role of these immunological findings. PMID- 16249288 TI - Activation of phospholipase A2 is associated with generation of placental lipid signals and fetal obesity. AB - CONTEXT: Obesity and diabetes during pregnancy are associated with increased insulin resistance and higher neonatal adiposity. In turn, insulin resistance triggers inflammatory pathways with accumulation of placental cytokines. OBJECTIVE: To determine placental signals that translate into development of excess adipose tissue, we investigated the role of phospholipases A2 (PLA2) as targets of inflammatory mediators. SETTING: The study was conducted at Case Western Reserve University, Department of Reproductive Biology. SUBJECTS: Volunteers gave informed written consent in accordance with the Institutional Review Board guidelines. Placenta and cord blood samples were obtained at the time of elective cesarean section in 15 term pregnancies. INTERVENTION: Neonatal anthropometric measurements were performed within 48 h of delivery. Placentas were grouped based on neonatal percentage body fat as obese (body fat > or = 16%) and lean control (body fat < or = 8%). MAIN OUTCOME MEASURES: The primary outcomes were placenta PLA2 expression and fatty acid concentration. RESULTS: Expression of PLA2G2A and PLA2G5, the main placenta phospholipases, was greater (P < 0.05) in placenta of obese compared with control neonates and was associated with increased 20:3 and 20:5 omega-3 polyunsaturated fatty acids. TNF-alpha and leptin content was increased 3-fold in placenta of obese neonates. TNF-alpha and leptin both induced a time-dependent activation of PLA2G2 and PLA2G5 in placental cells. CONCLUSION: Accumulation of omega-3 fatty acids through secretory PLA2 activation is associated with high neonatal adiposity. We propose that the generation of placental lipid mediators through TNF-alpha and leptin stimulation represents a key mechanism to favor excess fetal fat accretion. PMID- 16249289 TI - Effect of sex hormone administration on circulating ghrelin levels in peripubertal children. AB - BACKGROUND: Ghrelin levels gradually decrease throughout childhood and with advancing pubertal stage. The change during puberty is more pronounced in boys than girls. OBJECTIVE: The objective of the study was to investigate whether the pubertal drop in ghrelin secretion is modified by the increase in sex hormones. PATIENTS AND METHODS: Ghrelin levels were measured in 34 short peripubertal children (17 boys and 17 girls) aged 8-12.5 yr before and after sex hormone priming for GH stimulation testing. RESULTS: In boys, priming with testosterone increased testosterone to pubertal levels (23.7 +/- 7.1 nmol/liter), which in turn induced a marked decrease in ghrelin (from 1615.8 +/- 418.6 to 1390.0 +/- 352.0 pg/ml) and leptin (from 8.0 +/- 4.5 to 5.8 +/- 3.2 ng/ml) and an increase in IGF-I (from 162.7 +/- 52.8 to 291.1 +/- 101.6 ng/ml) (P < 0.001 for all parameters). In girls, priming with estrogen led to a supraphysiological increase in estradiol levels (1313.8 +/- 438.0 pmol/liter), which had no effect on ghrelin, leptin, or IGF-I. There was no correlation between ghrelin levels and levels of sex hormones, leptin, or body mass index in either boys or girls. CONCLUSIONS: A pharmacological increase in sex hormones is associated with a marked decline in circulating levels of ghrelin in boys but not girls. Additional longitudinal studies through puberty are needed to elucidate the physiological interaction between sex hormones and ghrelin. PMID- 16249290 TI - Endometriosis-specific genes identified by real-time reverse transcription polymerase chain reaction expression profiling of endometriosis versus autologous uterine endometrium. AB - CONTEXT: The etiology and molecular pathogenesis of endometriosis, a prevalent estrogen-dependent gynecologic disease, are poorly understood. OBJECTIVE: The objective of the study was to identify the differentially expressed genes between autologous ectopic and eutopic endometrium. DESIGN: Subtractive hybridization was used for a genome-wide search for differentially expressed genes between autologous ectopic and eutopic endometrium. Real-time RT-PCR was used for gene expression profiling in the paired tissue samples taken from multiple subjects. PATIENTS: The paired pelvic endometriosis and uterine endometrium tissue biopsies were procured from 15 patients undergoing laparoscopy or hysterectomy for endometriosis. RESULTS: Seventy-eight candidate genes were identified from the subtractive cDNA libraries. Seventy-six of these genes were investigated in approximately 8000 real-time PCR for their differential expression in 30 paired tissue biopsies from 15 patients affected by endometriosis. Cluster analysis on gene expression revealed highly consistent profiles in two groups of genes, despite the clinical heterogeneity of the 15 cases. Thirty-four genes specific to early disease point to their potential roles in establishment and evolution of endometriosis. Most interestingly, 14 genes were consistently dysregulated in the paired samples from the majority of the patients. Of these, there were two uncharacterized transcripts and two novel genes, and 10 were matched to known genes: IGFBP5, PIM2, RPL41, PSAP, FBLN1, SIPL, DLX5, HSD11B2, SET, and RHOE. CONCLUSIONS: Dysregulation of 14 genes was found to be overtly associated with endometriosis. Some of these genes, known to participate in estrogen activities and antiapoptosis, may play a role in the pathogenesis of endometriosis and may represent potential diagnostic markers or therapeutic targets for endometriosis. PMID- 16249293 TI - Founding a new College of Medicine at Florida State University. AB - In 2000, the Florida State University (FSU) College of Medicine was founded, becoming the first new allopathic medical school in the United States in over 20 years. The new medical school was to use community-based clinical training for the education of its students, create a technology-rich environment, and address primary care health needs of Florida's citizens, especially the elderly, rural, minorities, and underserved. The challenges faced during the creation of the new school, including accreditation and a leadership change, as well as accomplishments are described here. The new school admits a diverse student body made possible through its extensive outreach programs, fosters a humane learning environment through creation of student learning communities, has a distributed clinical training model-with clinical campuses in Orlando, Pensacola, Sarasota and Tallahassee, and with 70% of training occurring in ambulatory settings-and utilizes 21st-century information technology. The curriculum focuses on patient centered clinical training, using the biopsychosocial model of patient care throughout the entire medical curriculum, promotes primary care and geriatrics medicine through longitudinal community experiences, relies on a hybrid curriculum for delivery of the first two years of medical education with half of class sessions occurring in small groups and on a continuum of clinical skills development throughout the first three years, and uses an interdisciplinary departmental model for faculty, which greatly facilitates delivery of an integrated curriculum. The first class was admitted in 2001 and graduated in May 2005. In February 2005, the FSU College of Medicine received full accreditation from the Liaison Committee on Medical Education. PMID- 16249291 TI - Mature subcutaneous and visceral adipocyte concentrations of adiponectin are highly correlated in prepubertal children and inversely related to body mass index standard deviation score. AB - CONTEXT: Adiponectin is an adipocyte-specific protein with insulin-sensitizing properties. Several studies have examined the expression of adiponectin mRNA or tissue/secreted protein levels in fat obtained from adults, but none has assessed tissue levels in childhood. PATIENTS: Paired subcutaneous (Sc) and visceral (V) fat samples were obtained from 12 normal-weight children. MAIN OUTCOME MEASURES: Mature adipocytes were isolated and total adiponectin levels determined by ELISA. Insulin sensitivity and lipid parameters were assessed in fasting blood samples taken at the time of biopsy collection. RESULTS: A positive correlation was seen between the adiponectin concentration within the Sc and V mature adipocytes derived from each child (r = 0.924; P < 0.001). After logarithmic transformation of the Sc and V adiponectin concentrations (log-Sc and log-V) to render the data Gaussian, both log-Sc and log-V were found to be lower in those children with higher body mass index sd score (r = -0.621 and r = -0.357 respectively), although this reached statistical significance only in the Sc adipocytes (P = 0.03). Age was not related to either log-Sc or log-V adiponectin levels, although a significant negative association was seen with serum adiponectin (r = -0.589; P = 0.04). Log-Sc or log-V did not correlate with serum adiponectin concentrations, markers of insulin sensitivity, or circulating lipid levels. CONCLUSIONS: These data indicate a relationship between total adiponectin levels in different tissue compartments, suggesting either some form of interaction or coregulation by systemic factors, possibly related to body size/fat mass. Serum concentrations of total adiponectin were inversely related to age but showed no relationship with either tissue levels or body mass index sd score. PMID- 16249294 TI - Reinventing the academic health center. AB - Academic health centers have faced well-documented internal and external challenges over the last decade, putting pressure on organizational leaders to develop new strategies to improve performance while simultaneously addressing employee morale, patient satisfaction, educational outcomes, and research growth. In the aftermath of a failed merger, new leaders of The Pennsylvania State University College of Medicine and Milton S. Hershey Medical Center encountered a climate of readiness for a transformational change. In a case study of this process, nine critical success factors are described that contributed to significant performance improvement: performing a campus-wide cultural assessment and acting decisively on the results; making values explicit and active in everyday decisions; aligning corporate structure and governance to unify the academic enterprise and health system; aligning the next tier of administrative structure and function; fostering collaboration and accountability-the creation of unified campus teams; articulating a succinct, highly focused, and compelling vision and strategic plan; using the tools of mission-based management to realign resources; focusing leadership recruitment on organizational fit; and "growing your own" through broad-based leadership development. Outcomes assessment data for academic, research, and clinical performance showed significant gains between 2000 and 2004. Organizational transformation as a result of the nine factors is possible in other institutional settings and can facilitate a focus on crucial quality initiatives. PMID- 16249295 TI - The balancing act. PMID- 16249296 TI - Replacing the academic medical center's teaching hospital. AB - Addressing the need for updated teaching hospital facilities is one of the most significant issues that an academic medical center faces. The authors describe the process they underwent in deciding to build a new facility at the Medical University of South Carolina (MUSC). Initial issues included whether or not the teaching hospital would continue to play a role in clinical education and whether to replace or renovate the existing facility. Once the decision to build was reached, MUSC had to choose between an on-campus or distant site for the new hospital and determine what the function of the old hospital would be. The authors examine these questions and discuss the factors involved in different stages of decision making, in order to provide the academic medicine community guidance in negotiating similar situations. Open communication within MUSC and with the greater community was a key component of the success of the enterprise to date. The authors argue that decisions concerning site, size, and focus of the hospital must be made by developing university-wide and community consensus among many different constituencies. The most important elements in the success at MUSC were having unified leadership, incorporating constituent input, engaging an external consultant, remaining unfazed by unanticipated challenges, and adhering to a realistic, aggressive timetable. The authors share their strategies for identifying and successfully managing these complex and potentially divisive aspects of building a new teaching hospital. PMID- 16249297 TI - Implementing a simpler approach to mission-based planning in a medical school. AB - Changes in the education, research, and health care environments have had a major impact on the way in which medical schools fulfill their missions, and mission based management approaches have been suggested to link the financial information of mission costs and revenues with measures of mission activity and productivity. The authors describe a simpler system, termed Mission-Aligned Planning (MAP), and its development and implementation, during fiscal years 2002 and 2003, at the School of Medicine at the University of Texas Health Science Center at San Antonio, Texas. The MAP system merges financial measures and activity measures to allow a broad understanding of the mission activities, to facilitate strategic planning at the school and departmental levels. During the two fiscal years mentioned above, faculty of the school of medicine reported their annual hours spent in the four missions of teaching, research, clinical care, and administration and service in a survey designed by the faculty. A financial profit or loss in each mission was determined for each department by allocation of all departmental expenses and revenues to each mission. Faculty expenses (and related expenses) were allocated to the missions based on the percentage of faculty effort in each mission. This information was correlated with objective measures of mission activities. The assessment of activity allowed a better understanding of the real costs of mission activities by linking salary costs, assumed to be related to faculty time, to the missions. This was a basis for strategic planning and for allocation of institutional resources. PMID- 16249298 TI - Research centers and institutes in U.S. medical schools: a descriptive analysis. AB - Research centers and institutes are a common mechanism to organize and facilitate biomedical research at medical schools and universities. The authors report the results of a study on the size, scope, and range of activities of 604 research centers and institutes at research-intensive U.S. medical schools and their parent universities. Centers and institutes with primary missions of patient care, education, or outreach were not included. The findings indicate that, in addition to research, centers and institutes are involved in a range of activities, including education, service, and technology transfer. The centers and institutes the authors studied were more interdisciplinary than those included in previous studies on this topic. Most research centers and institutes did not have authority comparable to academic departments. Only 22% of centers directly appointed faculty members, and most center directors reported to a medical school dean or a department chair. A small group of centers and institutes ("power centers"), however, reported to a university president or provost, and may have considerable power and influence in academic decision making and resource allocation. Two main types of centers and institutes emerge from this research. The first type, which includes the vast of majority of centers, is modest in its scope and marginal in its influence. The second type- with greater amounts of funding, larger staffs, and direct access to institutional decisionmakers--may have a more significant role in the organization and governance of the medical school and university and in the ways that researchers interact within and across academic divisions. PMID- 16249299 TI - Promoting translational research in academic health centers: navigating the "roadmap". AB - The translation of hypothesis-driven research laboratory findings about basic disease mechanisms into clinically useful tests or therapies, particularly in pediatric diseases, is time-consuming, expensive, and not well supported by traditional research grant mechanisms. Accordingly, the development of new drugs and clinical assays has typically been largely the domain of the pharmaceutical industry. Aside from partnering with for-profit companies, academic health centers are challenged to find ways to actively engage in biomedical research to bridge the gap between basic and clinical research. The Translational Research Initiative (TRI) at Cincinnati Children's Hospital Medical Center was launched in 2001 with the mission to build an institutional infrastructure for promoting and facilitating the clinical implementation of investigator-initiated basic research. The TRI's goals are to provide grant support for proposals that are translational in nature and that address serious diagnostic or therapeutic deficiencies in pediatric illnesses; to create and support specialized research cores and a specialized office that provides support for research protocol development and regulatory affairs; and to organize educational opportunities focused on bridging communication between basic and clinical scientists and encouraging multidisciplinary interactions. The authors describe the program structure and provide an interim outcome report as measured by extramural funding obtained, Investigational New Drug applications filed, manuscripts published, clinical trials launched, and educational initiatives created. The broad success of this program suggests that it might serve as a model for other academic health centers in promoting and conducting translational research. PMID- 16249300 TI - Development of a data warehouse at an academic health system: knowing a place for the first time. AB - In 1998, the University of Michigan Health System embarked upon the design, development, and implementation of an enterprise-wide data warehouse, intending to use prioritized business questions to drive its design and implementation. Because of the decentralized nature of the academic health system and the development team's inability to identify and prioritize those institutional business questions, however, a bottom-up approach was used to develop the enterprise-wide data warehouse. Specific important data sets were identified for inclusion, and the technical team designed the system with an enterprise view and architecture rather than as a series of data marts. Using this incremental approach of adding data sets, institutional leaders were able to experience and then further define successful use of the integrated data made available to them. Even as requests for the use and expansion of the data warehouse outstrip the resources assigned for support, the data warehouse has become an integral component of the institution's information management strategy. The authors discuss the approach, process, current status, and successes and failures of the data warehouse. PMID- 16249301 TI - Managing knowledge and technology to foster innovation at the Ohio State University Medical Center. AB - Biomedical knowledge is expanding at an unprecedented rate-one that is unlikely to slow anytime in the future. While the volume and scope of this new knowledge poses significant organizational challenges, it creates tremendous opportunities to release and direct its power to the service of significant goals. The authors explain how the Center for Knowledge Management at The Ohio State University Medical Center, created during the academic year 2003-04, is doing just that by integrating numerous resource-intensive, technology-based initiatives-including personnel, services and infrastructure, digital repositories, data sets, mobile computing devices, high-tech patient simulators, computerized testing, and interactive multimedia-in a way that enables the center to provide information tailored to the needs of students, faculty and staff on the medical center campus and its surrounding health sciences colleges. The authors discuss how discovering, applying, and sharing new knowledge, information assets, and technologies in this way is a collaborative process. This process creates open ended opportunities for innovation and a roadmap for working toward seamless integration, synergy, and substantial enhancement of the academic medical center's research, educational, and clinical mission areas. PMID- 16249302 TI - Building research administration applications for the academic health center: a case study. AB - The academic health center information environment is saturated with information of varying quality and overwhelming quantity. The most significant challenge is transforming data and information into knowledge. The University of Cincinnati Medical Center's (UCMC) focus is to develop an information architecture comprising data structures, Web services, and user interfaces that enable individuals to manage the information overload so that they can create new knowledge. UCMC has accomplished much of what is reported in this article with the help of a four-year Integrated Advanced Information Management Systems (IAIMS) operation grant awarded by the National Library of Medicine in 2003. In the UCMC vision for knowledge management, individuals have reliable, secure access to information that is filtered, organized, and highly relevant for specific tasks and personal needs. Current applications and tool sets will evolve to become the next generation knowledge management applications or smart digital services. When smart digital services are implemented, silo applications will disappear. A major focus of UCMC's IAIMS grant is research administration. Testing and building out existing and new research administration applications and digital services is underway. The authors review UCMC's progress and results in developing a software architecture, tools, and services for research administration. Included are sections on the evolution to full integration, the impact of the work at UCMC to date, lessons learned during this research and development process, and future plans and needs. PMID- 16249303 TI - Developing effective interuniversity partnerships and community-based research to address health disparities. AB - Health disparities are an enormous challenge to American society. Addressing these disparities is a priority for U.S. society and especially for institutions of higher learning, with their threefold mission of education, service, and research. Collaboration across multiple intellectual disciplines will be critical as universities address health disparities. In addition, universities must collaborate with communities, with state partners, and with each other. Development of these collaborations must be sensitive to the history and unique characteristics of each academic institution and population. The authors describe the challenges of all three types of collaboration, but primarily focus on collaboration between research-intensive universities and historically black colleges and universities. The authors describe a four-year collaboration between Shaw University and the University of North Carolina at Chapel Hill (UNC-CH). These universities strategically developed multiple research initiatives to address health disparities, building on modest early success and personal relationships. These activities included participation by Shaw faculty in faculty development activities, multiple collaborative pilot studies, and joint participation in securing grants from the Agency for Health care Research and Quality of the federal Department of Health and Human Services and the National Institutes of Health, including a P-60 Project EXPORT center grant. These multiple activities were sometimes led by UNC-CH, sometimes by Shaw University. Open discussion of problems as they arose, realistic expectations, and mutual recognition of the strengths of each institution and its faculty have been critical in achieving successful collaboration to date. PMID- 16249304 TI - Building a transcontinental affiliation: a new model for academic health centers. AB - The recent affiliation of The Methodist Hospital (TMH) with Weill Medical College (WMC) of Cornell University and NewYork-Presbyterian Hospital is the first transcontinental primary affiliation between major, not-for-profit academic health centers (AHCs) in the United States. The authors describe the process followed, the issues involved, the initial accomplishments, and the opportunities envisioned. The key enablers of this affiliation were a rapid process, mutual trust based on existing professional relationships, and commitment to the project by Board leadership. Because of their geographic separation, the parties were not competitors in providing clinical care to their regional populations. The affiliation is nonexclusive, but is reciprocally primary in New York and Texas. Members of the TMH medical staff are eligible for faculty appointments at WMC. The principal areas of collaboration will be education, research, quality improvement, information technology, and international program development. The principal challenge has been the physical distance between the parties. Although extensive use of videoconferencing has been successful, personal contact is essential in establishing relationships. External processes impose a slower sequence and tempo of events than some might wish. This new model for AHCs creates exciting possibilities for the tripartite mission of research, education, and patient care. Realizing the potential of these opportunities will require unconstrained ideas and substantial investment of time and other critical resources. Since many consider that AHCs are in economic and cultural crisis, successful development of such possibilities could have importance beyond the collective interests of these three institutions. PMID- 16249305 TI - Effective organizational control: implications for academic medicine. AB - This article provides a framework for understanding the nature, role, functioning, design, and effects of organizational oversight systems. Using a case study with elements recognizable to an academic audience, the authors explore how a dean of a fictitious School of Medicine might use organizational control structures to develop effective solutions to global disarray within the academic medical center. Organizational control systems are intended to help influence the behavior of people as members of a formal organization. They are necessary to motivate people toward organizational goals, to coordinate diverse efforts, and to provide feedback about problems. The authors present a model of control to make this process more visible within organizations. They explore the overlap among academic medical centers and large businesses-for instance, each is a billion-dollar enterprise with complex internal and external demands and multiple audiences. The authors identify and describe how to use the key components of an organization's control system: environment, culture, structure, and core control system. Elements of the core control system are identified, described, and explored. These closely articulating elements include planning, operations, measurement, evaluation, and feedback systems. Use of control portfolios is explored to achieve goal-outcome congruence. Additionally, the authors describe how the components of the control system can be used synergistically by academic leadership to create organizational change, congruent with larger organizational goals. The enterprise of medicine is quickly learning from the enterprise of business. Achieving goal-action congruence will better position academic medicine to meet its multiple missions. PMID- 16249306 TI - 1962 banquet speech for the Nobel Prize in Literature: [excerpt]. PMID- 16249308 TI - A medical experience that taught me about humanism in medicine. PMID- 16249309 TI - A medical experience that taught me about humanism in medicine. PMID- 16249310 TI - Did recent changes in Medicare reimbursement hit teaching hospitals harder? AB - PURPOSE: To inform the policy debate on Medicare reimbursement by examining the financial effects of the Balanced Budget Act of 1997 (BBA) and subsequent adjustments on major academic medical centers, minor teaching hospitals, and nonteaching hospitals. METHOD: The authors simulated the impacts of BBA and subsequent BBA adjustments to predict the independent effects of changes in Medicare reimbursement on hospital revenues using 1997-2001 Medicare Cost Reports for all short-term acute-care hospitals in the United States. The authors also calculated actual (nonsimulated) operating and total margins among major teaching, minor teaching, and nonteaching hospitals to account for hospital response to the changes. RESULTS: The BBA and subsequent refinements reduced Medicare revenues to a greater degree in major teaching hospitals, but the fact that such hospitals had a smaller proportion of Medicare patients meant that the BBA reduced overall revenues by similar percentages across major, minor, and nonteaching hospitals. Consistently lower margins may have made teaching hospitals more vulnerable to cuts in Medicare support. CONCLUSIONS: Recent Medicare changes affected revenues at teaching and nonteaching hospitals more similarly than is commonly believed. However, the Medicare cuts under the BBA probably exacerbated preexisting financial strain on major teaching hospitals, and increased Medicare funding may not suffice to eliminate the strain. This report's findings are consistent with recent calls to support needed services of teaching hospitals through all-payer or general funds. PMID- 16249311 TI - Seasonally hibernating phenotype assessed through transcript screening. AB - Hibernation is a seasonally entrained and profound phenotypic transition to conserve energy in winter. It involves significant biochemical reprogramming, although our understanding of the underpinning molecular events is fragmentary and selective. We have conducted a large-scale gene expression screen of the golden-mantled ground squirrel, Spermophilus lateralis, to identify transcriptional responses associated specifically with the summer-winter transition and the torpid-arousal transition in winter. We used 112 cDNA microarrays comprising 12,288 probes that cover at least 5,109 genes. In liver, the profiles of torpid and active states in the winter were almost identical, although we identified 102 cDNAs that were differentially expressed between winter and summer, 90% of which were downregulated in the winter states. By contrast, in cardiac tissue, 59 and 115 cDNAs were elevated in interbout arousal and torpor, respectively, relative to the summer active condition, but only 7 were common to both winter states, and during arousal none was downregulated. In brain, 78 cDNAs were found to change in winter, 44 of which were upregulated. Thus transcriptional changes associated with hibernation are qualitatively modest and, since these changes are generally less than twofold, also quantitatively modest. Unbiased Gene Ontology profiling of the transcripts suggests a winter switch to beta-oxidation of lipids in liver and heart, a reduction in metabolism of toxic compounds and the urea cycle in liver, and downregulated electron transport in the brain. We identified just one strongly winter-induced transcript common to all tissues, namely an RNA-binding protein, RBM3. This analysis clearly differentiates responses of the principal tissues, identifies a large number of new genes undergoing regulation, and broadens our understanding of affected cellular processes that, in part, account for the winter-adaptive hibernating phenotype. PMID- 16249312 TI - Microarray screening of suppression subtractive hybridization-PCR cDNA libraries identifies novel RNAs regulated by dehydration in the rat supraoptic nucleus. AB - The magnocellular neurons (MCNs) of the supraoptic nucleus (SON) and paraventricular nucleus (PVN) of the hypothalamus are the principal site of biosynthesis of prepropeptide precursor of the antidiuretic hormone vasopressin (VP). This precursor is processed during anterograde axonal transportation to terminals in the posterior pituitary gland, where biologically active VP is stored until release into the general circulation in response to physiological activation of the SON by osmotic cues. By binding to V2-type receptors located in the kidney, VP decreases the amount of water lost in urine. Osmotic activation of the SON is accompanied by a dramatic morphological and functional remodeling. We have sought to understand the mechanistic basis of this plasticity in terms of the differential expression of genes. To identify such genes, we adopted an unbiased global approach based on suppressive subtractive hybridization polymerase chain reaction (SSH-PCR) Using this method, we generated libraries of clones putatively differentially expressed in control vs. dehydrated SON. To rapidly screen these libraries, 1,152 clones were subjected to microarray analysis, resulting in the identification of 459 differentially expressed transcripts. cDNA clones corresponding to 56 of these RNAs were sequenced, revealing many of them to be novel expressed sequence tags (ESTs). Four transcripts were shown by in situ hybridization (ISH) to be significantly up- or downregulated in the SON after dehydration. These genes may represent novel effectors or mediators of SON physiological remodeling. PMID- 16249313 TI - Genetic background affects cardiovascular responses to obstructive and simulated apnea. AB - We have recently demonstrated that genetic background significantly impacts the blood pressure and heart rate response to hypoxia (Campen MJ, Tagaito Y, Li J, Balbir A, Tankersley CG, Smith P, Schwartz A, and O'Donnell CP. Physiol Genomics 20: 15-20, 2005). Because hypoxia is considered a mediator of the acute and chronic cardiovascular complications of obstructive sleep apnea, we investigated whether genetic factors also influence the cardiovascular response to experimentally induced obstructive apnea (OA) and simulated apnea (SA). In three strains of inbred mice (C57BL/6J, DBA/2J, and FVB/J) anesthetized with urethane (1.2 g/kg), apnea was induced at end-expiration for 5- and 10-s periods in spontaneously breathing (OA) and mechanically ventilated (SA; pancuronium, 0.2 mg/kg bolus + 0.003 mg.kg(-1).min(-1)) animals before and after administration of an autonomic ganglionic blocker (hexamethonium, 20 mg/kg). In contrast to our previous findings with hypoxia, OA produced a marked hypertensive response in all three strains. However, strain impacted on the degree of bradycardia during OA, which was large in C57BL/6J and FVB/J mice and effectively absent in DBA/2J mice. In C57BL/6J but not FVB/J mice, the bradycardia was abolished with SA under mechanical ventilation. Cardiovascular responses to SA in all strains were eliminated by autonomic blockade. These data show that 1) DBA/2J mice, in contrast to the previous demonstration of marked bradycardia during hypoxia, unexpectedly do not produce bradycardia during apnea; 2) C57BL/6J mice exhibit a bradycardia that is dependent on input from thoracic afferents; and 3) FVB/J mice exhibit a bradycardia despite the loss of thoracic afferent input, consistent with a potent pressure response eliciting a baroreceptor-mediated bradycardia. Thus genetic background can affect both the pattern and magnitude of the cardiovascular response to apnea. PMID- 16249314 TI - Gene expression profiling during increased fetal lung expansion identifies genes likely to regulate development of the distal airways. AB - Growth and development of the fetal lungs is critically dependent on the degree to which the lungs are expanded by liquid; increases in fetal lung expansion accelerate lung growth, whereas reductions in lung expansion cause lung growth to cease. The mechanisms mediating expansion-induced lung growth are unknown but likely include alterations in the expression of genes that regulate lung cell proliferation. Our aim was to isolate and identify genes that are up- or downregulated by increased fetal lung expansion. In chronically catheterized fetal sheep at 126 days gestational age (GA), the left lung was expanded for 36 h, while the right lung remained at a control level of expansion. Subtraction hybridization was used to isolate genes differentially expressed between the left and right lungs. Screening of approximately 6,000 clones identified 1,138 and 118 cDNA fragments that were up- and downregulated by increased lung expansion, respectively. Northern blot analyses in separate groups of control fetuses and fetuses exposed to increased lung expansion were used to verify differential expression. Increased fetal lung expansion upregulated heat shock protein 47, thrombospondin-1, TROP2, tropoelastin, and tubulin-alpha3 in fetal lung tissue by approximately 200-300%; connective tissue growth factor and cysteine-rich angiogenic inducer 61 were increased by 20-30%. Genes downregulated by increased fetal lung expansion included CCSP-related protein-1, elongation factor-1alpha and vitamin D3 upregulated protein 1. We conclude that an increase in fetal lung expansion differentially regulates the expression of numerous genes in lung tissue, many of which have important putative roles in lung development, while the functions of others are currently unknown. PMID- 16249315 TI - Crucial role of the sarcoplasmic reticulum in the developmental regulation of Ca2+ transients and contraction in cardiomyocytes derived from embryonic stem cells. AB - In adult myocardium, excitation-contraction coupling is critically regulated by sarcoplasmic reticulum (SR) Ca2+ release via type 2 ryanodine receptor (RyR2), but generally, it is believed that SR-function is rudimentary in the fetal heart and in embryonic stem (ES) cell-derived cardiomyocytes (ESCMs), a possible source for cell replacement therapies. This study used wild-type (RyR2+/+) and RyR2 null (RyR2-/-) ESCMs as an in vitro model of cardiomyogenesis, together with pharmacological approaches and expression profiles of genes relevant for SR function, to elucidate the functional importance of RyR2 and SR on the regulation of Ca2+ transients and contraction during early cardiomyocyte development. During differentiation of RyR2+/+ ESCMs, SR function developed progressively with increased basal cytosolic free Ca2+ concentration ([Ca2+]i), enhanced frequency and amplitude, and decreased duration of Ca2+ transients that were inhibited by ryanodine and thapsigargin. These functional traits correlated with SR Ca2+ load and the expression of RyR2, SERCA2a, and phospholamban. RyR2-/- ESCMs, comparatively, demonstrated a significantly prolonged time-to-peak and reduced frequency of Ca2+ transients and contractions. Beta-adrenergic stimulation of RyR2+/+ ESCMs increased the frequency and amplitude of Ca2+ transients with differentiation but was much weaker in RyR2-/- ESCMs. We conclude that functional SR and control of RyR2-mediated SR Ca2+ release directly contribute to the spontaneous and beta-adrenergic receptor-stimulated contraction of ESCMs, even at very immature stages of development. PMID- 16249316 TI - Reduced Aph-1b expression causes tissue- and substrate-specific changes in gamma secretase activity in rats with a complex phenotype. AB - The gamma-secretase enzyme complex displays intramembrane catalytic activity toward many type I transmembrane proteins, including the Alzheimer-linked amyloid beta-protein precursor (APP) and the neuregulin receptor ErbB4. Active gamma secretase is a tetrameric protein complex consisting of presenilin-1 (or -2), nicastrin, PEN-2, and Aph-1a (or -1b). We have recently discovered that pharmacogenetically bred apomorphine-susceptible Wistar rats (APO-SUS) have only one or two copies of the Aph-1b gene (termed I/I and II/II rats, respectively), whereas their phenotypic counterparts (APO-UNSUS) have three copies (III/III). As a result, APO-SUS rats display reduced Aph-1b expression and a complex phenotype reminiscent of neurodevelopmental disorders. Here we determined in the I/I and III/III rats the gamma-secretase cleavage activity toward the three APP superfamily members, p75 neurotrophin receptor, ErbB4, and neuregulin-2, and found that the cleavage of only a subset of the substrates was changed. Furthermore, the observed differences were restricted to tissues that normally express relatively high Aph-1b compared with Aph-1a levels. Thus, we provide in vivo evidence that subtle alterations in gamma-secretase subunit composition may lead to a variety of affected (neuro)developmental signaling pathways and, consequently, a complex phenotype. PMID- 16249318 TI - A new ATS Committee: competing in the marketplace of ideas. PMID- 16249319 TI - Low testosterone in chronic obstructive pulmonary disease: does it really matter? PMID- 16249320 TI - Smoking: an injury with many lung manifestations. PMID- 16249321 TI - American Thoracic Society/Centers for Disease Control and Prevention/Infectious Diseases Society of America: controlling tuberculosis in the United States. AB - During 1993-2003, incidence of tuberculosis (TB) in the United States decreased 44% and is now occurring at a historic low level (14,874 cases in 2003). The Advisory Council for the Elimination of Tuberculosis has called for a renewed commitment to eliminating TB in the United States, and the Institute of Medicine has published a detailed plan for achieving that goal. In this statement, the American Thoracic Society (ATS), Centers for Disease Control and Prevention (CDC), and the Infectious Diseases Society of America (IDSA) propose recommendations to improve the control and prevention of TB in the United States and to progress toward its elimination. This statement is one in a series issued periodically by the sponsoring organizations to guide the diagnosis, treatment, control, and prevention of TB. This statement supersedes the previous statement by ATS and CDC, which was also supported by IDSA and the American Academy of Pediatrics (AAP). This statement was drafted, after an evidence-based review of the subject, by a panel of representatives of the three sponsoring organizations. AAP, the National Tuberculosis Controllers Association, and the Canadian Thoracic Society were also represented on the panel. This statement integrates recent scientific advances with current epidemiologic data, other recent guidelines from this series, and other sources into a coherent and practical approach to the control of TB in the United States. Although drafted to apply to TB-control activities in the United States, this statement might be of use in other countries in which persons with TB generally have access to medical and public health services and resources necessary to make a precise diagnosis of the disease; achieve curative medical treatment; and otherwise provide substantial science-based protection of the population against TB. This statement is aimed at all persons who advocate, plan, and work at controlling and preventing TB in the United States, including persons who formulate public health policy and make decisions about allocation of resources for disease control and health maintenance and directors and staff members of state, county, and local public health agencies throughout the United States charged with control of TB. The audience also includes the full range of medical practitioners, organizations, and institutions involved in the health care of persons in the United States who are at risk for TB. PMID- 16249322 TI - Poor choice of primary outcome in a clinical trial of pirfenidone in patients with IPF. PMID- 16249323 TI - The questionable efficacy of pirfenidone in IPF. PMID- 16249324 TI - Screening for chronic beryllium disease. PMID- 16249325 TI - Blood carbon monoxide will increase from a decline in pulmonary function alone. PMID- 16249326 TI - Molecular cloning, functional expression and subcellular localization of two putative vacuolar voltage-gated chloride channels in rice (Oryza sativa L.). AB - We isolated two cDNA clones (OsCLC-1 and OsCLC-2) homologous to tobacco CLC-Nt1, which encodes a voltage-gated chloride channel, from rice (Oryza sativa L. ssp. japonica, cv. Nipponbare). The deduced amino acid sequences were highly conserved (87.9% identity with each other). Southern blot analysis of the rice genomic DNA revealed that OsCLC-1 and OsCLC-2 were single-copy genes on chromosomes 4 and 2, respectively. OsCLC-1 was expressed in most tissues, whereas OsCLC-2 was expressed only in the roots, nodes, internodes and leaf sheaths. The level of expression of OsCLC-1, but not of OsCLC-2, was increased by treatment with NaCl. Both genes could partly substitute for GEF1, which encodes the sole chloride channel in yeast, by restoring growth under ionic stress. These results indicate that both genes are chloride channel genes. The proteins from both genes were immunochemically detected in the tonoplast fraction. Tagged synthetic green fluorescent protein which was fused to OsCLC-1 or OsCLC-2 localized in the vacuolar membranes. These results indicate that the proteins may play a role in the transport of chloride ions across the vacuolar membrane. We isolated loss-of function mutants of both genes from a panel of rice mutants produced by the insertion of a retrotransposon, Tos17, in the exon region, and found inhibition of growth at all life stages. PMID- 16249327 TI - A nuclear gene for the iron-sulfur subunit of mitochondrial complex II is specifically expressed during Arabidopsis seed development and germination. AB - Three nuclear genes, SDH2-1, SDH2-2 and SDH2-3, encode the essential iron-sulfur subunit of mitochondrial complex II in Arabidopsis thaliana. SDH2-1 and SDH2-2 probably arose via a recent duplication event and we reported that both are expressed in all organs from adult plants. In contrast, transcripts from SDH2-3 were not detected. Here we present data demonstrating that SDH2-3 is specifically expressed during seed development. SDH2-3 transcripts appear during seed maturation, persist through desiccation, are abundant in dry seeds and markedly decline during germination. Analysis of transgenic Arabidopsis plants carrying the SDH2-3 promoter fused to the beta-glucuronidase reporter gene shows that the SDH2-3 promoter is activated in the embryo during maturation, from the bent cotyledon stage. beta-Glucuronidase expression correlates with the appearance of endogenous SDH2-3 transcripts, suggesting that control of this nuclear gene is achieved through transcriptional regulation. Furthermore, progressive deletions of this promoter identified a 159 bp region (-223 to -65) important for SDH2-3 transcriptional activation in seeds. Interestingly, the SDH2-3 promoter remains active in embryonic tissues during germination and post-germinative growth, and is turned off in vegetative tissues (true leaves). In contrast to SDH2-3 transcripts, SDH2-1 and SDH2-2 transcripts are barely detected in dry seeds and increase during germination and post-germinative growth. The opposite expression patterns of SDH2 nuclear genes strongly suggest that during germination the embryo-specific SDH2-3 is replaced by SDH2-1 or SDH2-2 in mitochondrial complex II. PMID- 16249328 TI - Adaptation and incipient sympatric speciation of Bacillus simplex under microclimatic contrast at "Evolution Canyons" I and II, Israel. AB - The microevolutionary dynamics of prokaryotes in natural habitats, such as soil, is poorly understood in contrast to our increasing knowledge on their immense diversity. We performed microevolutionary analyses on 945 soil isolates of Bacillus simplex from "Evolution Canyons" I (Carmel, Israel) and II (Galilee, Israel). These canyons represent similar ecological replicates, separated by 40 km, with highly contrasting interslope abiotic and biotic conditions in each (within a distance of only 100-400 m). Strains representing genetic groups were identical in their 16S sequences, suggesting high genetic similarity and monophyletic origin. Parallel and nested phylogenetic structures correlated with ecological contrasts rather than geographical distance. Additionally, slope specific populations differed substantially in their diversity. The levels of DNA repair (determined by UV sensitivity) and spontaneous mutation rate (resistance to rifampicin) relate to ecological stress and phylogeny. Altogether, the results suggest adaptive radiation at a microscale. We discuss the observed adaptive population structures in the context of incipient sympatric speciation in soil bacteria. We conclude that, despite different biology, prokaryotes, like sexually reproducing eukaryotes, may consist of true species and parallel ecological speciation in eukaryotes. PMID- 16249329 TI - Spatial regulation of microRNA gene expression in the Drosophila embryo. AB - MicroRNAs (miRNAs) regulate posttranscriptional gene activity by binding to specific sequences in the 3' UTRs of target mRNAs. A number of metazoan miRNAs have been shown to exhibit tissue-specific patterns of expression. Here, we investigate the possibility that localized expression is mediated by tissue specific enhancers, comparable to those seen for protein-coding genes. Two miRNA loci in Drosophila melanogaster are investigated, the mir-309-6 polycistron (8 miR) and the mir-1 gene. The 8-miR locus contains a cluster of eight distinct miRNAs that are transcribed in a common precursor RNA. The 8-miR primary transcript displays a dynamic pattern of expression in early embryos, including repression at the anterior and posterior poles. An 800-bp 5' enhancer was identified that recapitulates this complex pattern when attached to a RNA polymerase II core promoter fused to a lacZ-reporter gene. The miR-1 locus is specifically expressed in the mesoderm of gastrulating embryos. Bioinformatics methods were used to identify a mesoderm-specific enhancer located approximately 5 kb 5' of the miR-1 transcription unit. Evidence is presented that the 8-miR enhancer is regulated by the localized Huckebein repressor, whereas miR-1 is activated by Dorsal and Twist. These results provide evidence that restricted activities of the 8-miR and miR-1 miRNAs are mediated by classical tissue specific enhancers. PMID- 16249330 TI - The search for glycan function: fucosylation of the TGF-beta1 receptor is required for receptor activation. PMID- 16249331 TI - A geometric construction determines all permissible strand arrangements of sandwich proteins. AB - For a large class of proteins called sandwich-like proteins (SPs), the secondary structures consist of two beta-sheets packed face-to-face, with each beta-sheet consisting typically of three to five beta-strands. An important step in the prediction of the three-dimensional structure of a SP is the prediction of its supersecondary structure, namely the prediction of the arrangement of the beta strands in the two beta-sheets. Recently, significant progress in this direction was made, where it was shown that 91% of observed SPs form what we here call "canonical motifs." Here, we show that all canonical motifs can be constructed in a simple manner that is based on thermodynamic considerations and uses certain geometric structures. The number of these structures is much smaller than the number of possible strand arrangements. For instance, whereas for SPs consisting of six strands there exist a priori 900 possible strand arrangements, there exist only five geometric structures. Furthermore, the few motifs that are noncanonial can be constructed from canonical motifs by a simple procedure. PMID- 16249332 TI - Profile of Stephen H. Schneider. PMID- 16249333 TI - Multiple modes of interaction of the deglycosylation enzyme, mouse peptide N glycanase, with the proteasome. AB - Peptide N-glycanase (PNGase) is involved in the cleavage of oligosaccharide chains from misfolded glycoproteins that are destined for degradation by the proteasome. Earlier, a number of potential binding partners of mouse PNGase (mPNGase) were detected by using the yeast two-hybrid system. In the current study, an in vitro system was set up to investigate direct interactions between mPNGase and these candidate proteins. Although the yeast two-hybrid system suggested an interaction of six different proteins with mPNGase, only mHR23B and the proteasome subunit mS4 were found to interact with mPNGase. In fact, mS4 competes with mHR23B for binding to mPNGase. These results suggested two possible pathways for the interaction between mPNGase and the proteasome. In one pathway, mHR23B mediates the interaction between mPNGase and the proteasome. In an alternative pathway, mPNGase directly binds to the proteasome subunit, mS4. In either case, it is clear that PNGase is located in close proximity to the proteasome and is available for deglycosylation of glycoproteins destined for degradation. Surprisingly, mPNGase also was found to mediate binding of the cytoplasmic protein, p97, to the proteasome through the formation of a ternary complex made up of mHR23B, mPNGase, and p97. Because p97 is known to bind to the endoplasmic reticulum membrane protein AMFR (gp78), an E3 ligase, we propose a model in which p97, mPNGase, and mHR23B mediate interaction of the endoplasmic reticulum with the proteasome. PMID- 16249334 TI - Profile of Deborah P. Delmer. PMID- 16249335 TI - Crystal structure of the Bacillus subtilis anti-alpha, global transcriptional regulator, Spx, in complex with the alpha C-terminal domain of RNA polymerase. AB - Spx, a global transcription regulator in Bacillus subtilis, interacts with the C terminal domain of the alpha subunit (alphaCTD) of RNA polymerase to control gene expression under conditions of disulfide stress, which is sensed by disulfide bond formation between Spx residues C10 and C13. Here, we describe the crystal structure of the B. subtilis alphaCTD bound to oxidized Spx. Analysis of the complex reveals interactions between three regions of "anti-alpha" Spx and helix alpha1 and the "261" determinant of alphaCTD. The former contact could disrupt the interaction between alphaCTD and activator proteins or alter the DNA-bound conformation of alphaCTD, thereby repressing activator-stimulated transcription. Binding to the 261 determinant would prevent interaction between alphaCTD and region 4 of sigma(A). Intriguingly, the Spx disulfide bond is far from the alphaCTD-Spx interface, suggesting that Spx regulates transcription allosterically or through the redox-dependent creation or destruction of binding sites for additional components of the transcription machinery. PMID- 16249337 TI - The role of science in solving the world's emerging water problems. PMID- 16249336 TI - Recruitment of governing elements for electron transfer in the nitric oxide synthase family. AB - At least three building blocks are responsible for the molecular basis of the modulation of electron transfer in nitric oxide synthase (NOS) isoforms: the calmodulin-binding sequence, the C-terminal extension, and the autoregulatory loop in the reductase domain. We have attempted to impart the control conferred by the C termini of NOS to cytochrome P450 oxidoreductase (CYPOR), which contains none of these regulatory elements. The effect of these C termini on the properties of CYPOR sheds light on the possible evolutionary origin of NOS and addresses the recruitment of new peptides on the development of new functions for CYPOR. The C termini of NOSs modulate flavoprotein-mediated electron transfer to various electron acceptors. The reduction of the artificial electron acceptors cytochrome c, 2,6-dichlorophenolindophenol, and ferricyanide was inhibited by the addition of any of these C termini to CYPOR, whereas the reduction of molecular O(2) was increased. This suggests a shift in the rate-limiting step, indicating that the NOS C termini interrupt electron flux between flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD) and/or the electron acceptors. The modulation of CYPOR by the addition of the NOS C termini is also supported by flavin reoxidation and fluorescence-quenching studies and antibody recognition of the C-terminal extension. These experiments support the origin of the NOS enzymes from modules consisting of a heme domain and CYPOR or ferredoxin-NADP(+) reductase- and flavodoxin-like subdomains that constitute CYPOR, followed by further recruitment of smaller modulating elements into the flavin-binding domains. PMID- 16249338 TI - Central role of a bacterial two-component gene regulatory system of previously unknown function in pathogen persistence in human saliva. AB - The molecular genetic mechanisms used by bacteria to persist in humans are poorly understood. Group A Streptococcus (GAS) causes the majority of bacterial pharyngitis cases in humans and is prone to persistently inhabit the upper respiratory tract. To gain information about how GAS survives in and infects the oropharynx, we analyzed the transcriptome of a serotype M1 strain grown in saliva. The dynamic pattern of changes in transcripts of genes [spy0874/0875, herein named sptR and sptS (sptR/S), for saliva persistence] encoding a two component gene regulatory system of unknown function suggested that SptR/S contributed to persistence of GAS in saliva. Consistent with this idea, an isogenic nonpolar mutant strain (DeltasptR) was dramatically less able to survive in saliva compared with the parental strain. Iterative expression microarray analysis of bacteria grown in saliva revealed that transcripts of several known and putative GAS virulence factor genes were decreased significantly in the DeltasptR mutant strain. Compared with the parental strain, the isogenic mutant strain also had altered transcripts of multiple genes encoding proteins involved in complex carbohydrate acquisition and utilization pathways. Western immunoblot analysis and real-time PCR analysis of GAS in throat swabs taken from humans with pharyngitis confirmed the findings. We conclude that SptR/S optimizes persistence of GAS in human saliva, apparently by strategically influencing metabolic pathways and virulence factor production. The discovery of a genetic program that significantly increased persistence of a major human pathogen in saliva enhances understanding of how bacteria survive in the host and suggests new therapeutic strategies. PMID- 16249339 TI - Pervasive social deficits, but normal parturition, in oxytocin receptor-deficient mice. AB - The oxytocin receptor (OXTR) and its ligand, oxytocin (OXT), regulate reproductive physiology (i.e., parturition and lactation) and sociosexual behaviors. To define the essential functions of OXTR, we generated mice with a null mutation in the Oxtr gene (Oxtr(-/-)) and compared them with OXT-deficient (Oxt(-/-)) mice. Oxtr(-/-) mice were viable and had no obvious deficits in fertility or reproductive behavior. Oxtr(-/-) dams exhibited normal parturition but demonstrated defects in lactation and maternal nurturing. Infant Oxtr(-/-) males emitted fewer ultrasonic vocalizations than wild-type littermates in response to social isolation. Adult Oxtr(-/-) males also showed deficits in social discrimination and elevated aggressive behavior. Ligand Oxt(-/-) males from Oxt(-/-) dams, but not from Oxt(+/-) dams, showed similar high levels of aggression. These data suggest a developmental role for the OXT/OXTR system in shaping adult aggressive behavior. Our studies demonstrate that OXTR plays a critical role in regulating several aspects of social behavior and may have important implications for developmental psychiatric disorders characterized by deficits in social behavior. PMID- 16249340 TI - Probing the active site tightness of DNA polymerase in subangstrom increments. AB - We describe the use of a series of gradually expanded thymine nucleobase analogs in probing steric effects in DNA polymerase efficiency and fidelity. In these nonpolar compounds, the base size was increased incrementally over a 1.0-A range by use of variably sized atoms (H, F, Cl, Br, and I) to replace the oxygen molecules of thymine. Kinetics studies with DNA Pol I (Klenow fragment, exonuclease-deficient) in vitro showed that replication efficiency opposite adenine increased through the series, reaching a peak at the chlorinated compound. Efficiency then dropped markedly as a steric tightness limit was apparently reached. Importantly, fidelity also followed this trend, with the fidelity maximum at dichlorotoluene, the largest compound that fits without apparent repulsion. The fidelity at this point approached that of wild-type thymine. Surprisingly, the maximum fidelity and efficiency was found at a base pair size significantly larger than the natural size. Parallel bypass and mutagenesis experiments were then carried out in vivo with a bacterial assay for replication. The cellular results were virtually the same as those seen in solution. The results provide direct evidence for the importance of a tight steric fit on DNA replication fidelity. In addition, the results suggest that even high-fidelity replicative enzymes have more steric room than necessary, possibly to allow for an evolutionarily advantageous mutation rate. PMID- 16249341 TI - Vacuolar Na+/H+ antiporter cation selectivity is regulated by calmodulin from within the vacuole in a Ca2+- and pH-dependent manner. AB - The selective movement of ions between intracellular compartments is fundamental for eukaryotes. Arabidopsis thaliana Na(+)/H(+) exchanger 1 (AtNHX1), the most abundant vacuolar Na(+)/H(+) antiporter in A. thaliana, has important roles affecting the maintenance of cellular pH, ion homeostasis, and the regulation of protein trafficking. Previously, we have shown that the AtNHX1 C-terminal hydrophilic region localized in the vacuolar lumen plays an important role in regulating the antiporter's activity. Here, we have identified A. thaliana calmodulin-like protein 15 (AtCaM15), which interacts with the AtNHX1 C terminus. When expressed in yeast, AtCaM15 is localized in the vacuolar lumen. The transient expression of AtCaM15 in Arabidopsis leaf protoplasts showed that AtCaM15 is present in the central vacuole. The binding of AtCaM15 to AtNHX1 was Ca(2+)- and pH-dependent and decreased with increasing pH values. Our results also show that the binding of AtCaM15 to AtNHX1 modified the Na(+)/K(+) selectivity of the antiporter, decreasing its Na(+)/H(+) exchange activity. Taken together, the presence of a vacuolar calmodulin-like protein acting on the vacuolar-localized AtNHX1 C terminus in a Ca(2+)- pH-dependent manner suggests the presence of signaling entities acting within the vacuole. PMID- 16249342 TI - Distinctions in the specificity of E2F function revealed by gene expression signatures. AB - The E2F family of transcription factors provides essential activities for coordinating the control of cellular proliferation and cell fate. Both E2F1 and E2F3 proteins have been shown to be particularly important for cell proliferation, whereas the E2F1 protein has the capacity to promote apoptosis. To explore the basis for this specificity of function, we used DNA microarray analysis to probe for the distinctions in the two E2F activities. Gene expression profiles that distinguish either E2F1- or E2F3-expressing cells from quiescent cells are enriched in genes encoding cell cycle and DNA replication activities, consistent with many past studies. E2F1 profile is also enriched in genes known to function in apoptosis. We also identified patterns of gene expression that specifically differentiate the activity of E2F1 and E2F3; this profile is enriched in genes known to function in mitosis. The specificity of E2F function has been attributed to protein interactions mediated by the marked box domain, and we now show that chimeric E2F proteins generate expression signatures that reflect the origin of the marked box, thus linking the biochemical mechanism for specificity of function with specificity of gene activation. PMID- 16249343 TI - Synergy between interstitial flow and VEGF directs capillary morphogenesis in vitro through a gradient amplification mechanism. AB - Cell organization is largely orchestrated by extracellular gradients of morphogenetic proteins. VEGF, an essential factor for capillary formation, is stored in the extracellular matrix, but the mechanisms by which it and other matrix-bound morphogens are mobilized to form spatial gradients are poorly understood. Here, we suggest an efficient mechanism for morphogen gradient generation by subtle biophysical forces in an in vitro model of capillary morphogenesis. Using a fibrin-bound VEGF variant that is released proteolytically to mimic the in vivo situation, we report that low levels of interstitial flow act synergistically with VEGF to drive endothelial organization, whereas each stimulus alone has very little effect. To help account for this synergy, we show how these slow flows can bias the distribution of cell-secreted proteases, which leads, interestingly, to the creation of an increasing VEGF gradient relative to the cell and skewed in the direction of flow. In contrast, diffusion alone can only account for symmetric, decreasing autocrine gradients. Indeed, branching of capillary structures was biased in the direction of flow only with the combination of VEGF and flow. This work thus demonstrates a general mechanism of morphogen gradient generation and amplification by small ubiquitous mechanical forces that are known to exist in vivo. PMID- 16249344 TI - Simulating movement of tRNA into the ribosome during decoding. AB - Decoding is the key step during protein synthesis that enables information transfer from RNA to protein, a process critical for the survival of all organisms. We have used large-scale (2.64 x 10(6) atoms) all-atom simulations of the entire ribosome to understand a critical step of decoding. Although the decoding problem has been studied for more than four decades, the rate-limiting step of cognate tRNA selection has only recently been identified. This step, known as accommodation, involves the movement inside the ribosome of the aminoacyl-tRNA from the partially bound "A/T" state to the fully bound "A/A" state. Here, we show that a corridor of 20 universally conserved ribosomal RNA bases interacts with the tRNA during the accommodation movement. Surprisingly, the tRNA is impeded by the A-loop (23S helix 92), instead of enjoying a smooth transition to the A/A state. In particular, universally conserved 23S ribosomal RNA bases U2492, C2556, and C2573 act as a 3D gate, causing the acceptor stem to pause before allowing entrance into the peptidyl transferase center. Our simulations demonstrate that the flexibility of the acceptor stem of the tRNA, in addition to flexibility of the anticodon arm, is essential for tRNA selection. This study serves as a template for simulating conformational changes in large (>10(6) atoms) biological and artificial molecular machines. PMID- 16249345 TI - Inhibition of colony-stimulating-factor-1 signaling in vivo with the orally bioavailable cFMS kinase inhibitor GW2580. AB - Colony-stimulating-factor-1 (CSF-1) signaling through cFMS receptor kinase is increased in several diseases. To help investigate the role of cFMS kinase in disease, we identified GW2580, an orally bioavailable inhibitor of cFMS kinase. GW2580 completely inhibited human cFMS kinase in vitro at 0.06 microM and was inactive against 26 other kinases. GW2580 at 1 microM completely inhibited CSF-1 induced growth of mouse M-NFS-60 myeloid cells and human monocytes and completely inhibited bone degradation in cultures of human osteoclasts, rat calvaria, and rat fetal long bone. In contrast, GW2580 did not affect the growth of mouse NS0 lymphoblastoid cells, human endothelial cells, human fibroblasts, or five human tumor cell lines. GW2580 also did not affect lipopolysaccharide (LPS)-induced TNF, IL-6, and prostaglandin E2 production in freshly isolated human monocytes and mouse macrophages. After oral administration, GW2580 blocked the ability of exogenous CSF-1 to increase LPS-induced IL-6 production in mice, inhibited the growth of CSF-1-dependent M-NFS-60 tumor cells in the peritoneal cavity, and diminished the accumulation of macrophages in the peritoneal cavity after thioglycolate injection. Unexpectedly, GW2580 inhibited LPS-induced TNF production in mice, in contrast to effects on monocytes and macrophages in vitro. In conclusion, GW2580's selective inhibition of monocyte growth and bone degradation is consistent with cFMS kinase inhibition. The ability of GW2580 to chronically inhibit CSF-1 signaling through cFMS kinase in normal and tumor cells in vivo makes GW2580 a useful tool in assessing the role of cFMS kinase in normal and disease processes. PMID- 16249346 TI - Weekly administration of docetaxel and paclitaxel in metastatic or advanced breast cancer. AB - The taxanes docetaxel (Taxotere; Aventis Pharmaceuticals Inc., Bridgewater, NJ, http://www.aventispharma-us.com) and paclitaxel (Taxol; Bristol-Myers Squibb, Princeton, NJ, http://www.bms.com) have significant clinical activity in metastatic breast cancer. A number of clinical trials have evaluated the tolerability and efficacy of weekly taxane administration to optimize the benefit to-risk ratio in metastatic breast cancer. Single-agent studies with docetaxel and paclitaxel in metastatic breast cancer show clinically significant antitumor activity even in advanced, heavily pretreated, resistant, and/or refractory disease. This activity is also evident with taxane-based combination regimens. Severe hematologic and nonhematologic toxicities are infrequent, with other toxicities noted based on the dose and weekly regimen selected. Weekly docetaxel and paclitaxel regimens represent valuable therapeutic options for women with metastatic breast cancer and have entered evaluation as part of adjuvant therapy for this disease. PMID- 16249347 TI - Hematologic safety and tolerability of topotecan in recurrent ovarian cancer and small cell lung cancer: an integrated analysis. AB - The purpose was to conduct an integrated analysis of the cumulative hematologic toxicity of topotecan in patients with relapsed ovarian cancer and small cell lung cancer (SCLC). Data were pooled from eight phase II and phase III clinical studies performed in patients with relapsed stage III/IV ovarian cancer or extensive SCLC treated with topotecan at a dose of 1.5 mg/m(2) per day on days 1 5 of a 21-day course. Quantitative hematologic toxicities were assessed using the National Cancer Institute Common Toxicity Criteria. A total of 4,124 courses of therapy was administered to the 879 patients in the pooled population. Grade 4 neutropenia was experienced by 78% of patients. The lowest nadirs for neutrophils and platelets generally occurred after the first course of therapy, followed by improvement or stabilization in subsequent courses. Neutropenia was noncumulative. During the first course, significant risk factors were identified: renal impairment and advanced age (grade 3/4 thrombocytopenia and grade 4 neutropenia) and prior radiotherapy; performance status score > or =2; SCLC; and exposure to both cisplatin (Platinol; Bristol-Myers Squibb, Princeton, NJ, http://www.bms.com) and carboplatin (Paraplatin; Bristol-Myers Squibb) (grade 3/4 thrombocytopenia only). The most frequent interventions for hematologic toxicities were RBC transfusions, treatment delays, G-CSF support, and dose reductions. Analysis of neutrophil and platelet nadirs and dosing for each course of therapy showed no apparent evidence of cumulative neutropenia or thrombocytopenia. The risk of grade 3 or 4 anemia was higher during the first four courses of therapy and may need to be more aggressively managed with erythropoietin therapy. PMID- 16249348 TI - Topotecan as second-line therapy for ovarian cancer: dosage versus toxicity. AB - In this issue of The Oncologist, Armstrong et al. present an analysis of the use of topotecan (Hycamtin; GlaxoSmithKline, Philadelphia, http://www.gsk.com) in the second-line treatment of both ovarian cancer and small cell carcinoma of the lung. This cytotoxic agent has clearly been demonstrated to be a useful drug in a population of patients with both of these conditions. However, the description of the nature of the toxicity, as stated in the manuscript, must be questioned along with comments made regarding the relative toxicity of alternative cytotoxic agents frequently used in similar settings. The purpose of this discussion absolutely is not to negate the unquestioned, demonstrated usefulness of topotecan as second-line therapy in ovarian cancer but rather to point out that the U.S. Food and Drug Administration-approved dose level of 1.5 mg/m(2) per day x 5 days can cause substantial and highly clinically relevant bone marrow toxicity. Whether this toxicity, which can result in a level of fatigue that may cause responding patients to discontinue treatment, should simply be labeled "excessive" rather than "cumulative" appears to be a matter of semantics rather than an important distinction. Whether delivery of a lower dose of topotecan (1 mg/m(2)-1.25 mg/m(2) per day x 5 days) will essentially eliminate concern for the development of severe clinically relevant marrow toxicity is uncertain, but the risk will certainly be substantially reduced. PMID- 16249349 TI - Topotecan: weighing in when there are many options. AB - The manuscript by Armstrong et al. in this issue of The Oncologist, reviewing the toxicity and efficacy of full-dose topotecan delivered as second-line therapy in individuals with small cell lung cancer or ovarian cancer, will likely serve as both the most comprehensive and the final review of this active agent delivered at the U.S. Food and Drug Administration-approved dosage and schedule. The review represents large, relative lyhomogeneous patient populations with prior platinum exposure and convincingly describes topotecan as an agent with activity that is comparable with those of all other approved drugs in this setting and a well defined and relatively circumscribed set of toxicities. While the activity of topotecan in small cell carcinoma provides individuals with the hope for further palliation at the time of tumor recurrence, as discussed by Dr. Markman in this issue, the best use of this agent in the management of recurrent ovarian cancer is far from clear because, in part, of a growing list of approved active agents, most notably liposomal doxorubicin, gemcitabine, and weekly paclitaxel; a return to platinum; or a host of other agents such as vinorelbine, altretamine, irinotecan, docetaxel, etoposide, and others. The most pressing need is to accept that palliative therapies are designed to palliate and improve symptoms, and to move away from end points of radiologic and marker response to a focus on "clinical benefit" and clinically more meaningful end points. PMID- 16249350 TI - Antibody-based therapies for colorectal cancer. AB - The recent successful development of novel monoclonal antibodies that target key components of biologic pathways has expanded the armamentarium of treatment options for patients with colorectal cancer. Two targets in particular--the process of new blood vessel development, or angiogenesis, and the epidermal growth factor receptor and its signaling pathway--are exploited by the newest monoclonal antibodies that are available for use in colorectal cancer patients. This clinical review focuses on the defining role of the two most clinically advanced novel agents, bevacizumab (Avastin; Genentech, Inc., South San Francisco, CA, http://www.gene.com) and cetuximab (Erbitux; ImClone Systems, Inc., New York, http://www.imclone.com), in colorectal cancer. PMID- 16249351 TI - Combined therapy with weekly irinotecan, infusional 5-fluorouracil and the selective COX-2 inhibitor rofecoxib is a safe and effective second-line treatment in metastatic colorectal cancer. AB - The purpose of this study was to determine the tolerability and activity of rofecoxib (Vioxx; Merck & Co., Inc., Whitehouse Station, NJ, http://www.merck.com) combined with weekly irinotecan (Camptosar; Pfizer Pharmaceuticals, New York, http://www.pfizer.com) and infusional 5-fluorouracil (5-FU) as second-line therapy in metastatic colorectal cancer (MCRC). Enrolled patients had previously treated metastatic disease, were aged > or =18 to < or =75 years, and had adequate performance status. A cycle of treatment consisted of i.v. irinotecan on days 1, 8, 15, and 22, rofecoxib at an oral dose of 50 mg/day, and infusional 5-FU at a fixed dose of 200 mg/m(2) per day for 5 weeks followed by 3 weeks of therapy with rofecoxib alone. In the dose-finding study, the starting dose of irinotecan was 87.5 mg/m(2) and further dose escalations were planned by increments of 12.5 mg/m(2) up to 125 mg/m(2). Forty-eight consecutive patients were enrolled in the study. Among the 15 cases enrolled in the dose finding study, one patient experienced grade 3 reversible diarrhea as the dose limiting toxicity, at the fourth dose level tested. Therefore, the dose of irinotecan for the phase II study was 125 mg/m(2), and 33 patients were enrolled and received a total of 75 cycles. Hematological side effects were moderate, with grade 4 neutropenia recorded in only two patients. The most common nonhematological toxicity was diarrhea, occurring in 25 patients (75.8%) and considered to be of grade 3 in 12 patients (36.4%). Sixteen patients achieved partial responses (48.5%; 95% confidence interval [CI], 30.8%-66.5%), and another 10 patients (30.3%) had stable disease. The median time to progression was 7 months (95% CI, 5-12) and the median overall survival (OS) was 18 months; the 1 year estimated OS rate was 69.4%. The unique schedule tested in this study is feasible, is well-tolerated, and has promising activity in patients with MCRC after progression on oxaliplatin (Eloxatin; Sanofi-Synthelabo Inc., New York, http://www.sanofi-synthelabo.us)-based chemotherapy. PMID- 16249352 TI - Thalidomide in advanced hepatocellular carcinoma with optional low-dose interferon-alpha2a upon progression. AB - PURPOSE: To evaluate thalidomide in advanced hepatocellular carcinoma (HCC) and to evaluate combined thalidomide and low-dose interferon-alpha2a (IFN-alpha2a) after tumor progression on thalidomide. Systemic therapy is minimally effective in HCC and tumor angiogenesis is a potential therapeutic target. PATIENTS AND METHODS: Patients with unresectable HCC were eligible if they had preserved hepatic and renal function. The initial thalidomide dosage was 200 mg daily and was adjusted for toxicity. Upon progression, patients could continue thalidomide with additional low-dosage (one million units twice daily) IFN-alpha2a. RESULTS: Thirty-eight enrolled patients were predominantly hepatitis C virus infected (53%), Child-Pugh class A (79%), and Eastern Cooperative Oncology Group performance status 0-1 (92%); 60% had extrahepatic metastasis. Confirmed disease control was seen in seven patients (18%) and included one complete and one partial response (5% response rate). The median progression-free survival was 2.1 months, and median overall survival was 5.5 months. Tumor invasion of the portal vein or vena cava, large (>10 cm) tumor, and younger age were associated with shorter overall survival. Toxicity included fatigue in 74% of patients. Six patients stopped therapy because of side effects, including two patients (5%) with grade 4 arteriothrombotic events. Five patients continued thalidomide upon progression with the addition of IFN-alpha2a; there was no disease control and 80% had grade 3 toxicity. CONCLUSIONS: Thalidomide is not well tolerated and confers limited disease control in advanced HCC. Combination thalidomide and low dose IFN-alpha2a is neither safe nor efficacious in this population. PMID- 16249353 TI - Paclitaxel/carboplatin/etoposide versus paclitaxel/topotecan for extensive-stage small cell lung cancer: a Minnie Pearl Cancer Research Network randomized, prospective phase II trial. AB - PURPOSE: To compare the combination of paclitaxel (Taxol; Bristol-Myers Squibb, Princeton, NJ, http://www.bms.com) and topotecan (Hycamtin; Glaxo SmithKline, Philadelphia, http://www.gsk.com) with paclitaxel, carboplatin (Paraplatin; Bristol-Myers Squibb), and etoposide (Etopophos, VePesid; Bristol-Myers Squibb) in patients with previously untreated extensive-stage small cell lung cancer. PATIENTS AND METHODS: In this phase II trial, 120 patients were randomly allocated to receive either topotecan (1.5 mg/m(2) i.v. days 1, 2, and 3) and paclitaxel (175 mg/m(2) i.v. day 1) every 21 days orpaclitaxe l (200 mg/m(2) i.v. day 1), carboplatin (area under the concentration-time curve 6 i.v. day 1), and etoposide (50 mg/100 mg alternating daily by mouth days 1-10) every 21 days, each regimen for a maximum of eight cycles. The primary end points were objective response rate and time to progression. RESULTS: The paclitaxel-carboplatin etoposide combination produced a significantly higher overall response rate (78% versus 48%), longer median time to progression (7.6 months versus 5.5 months), and greater number of patients free from progression at 1 year (14% versus 8%) compared with paclitaxel plus topotecan. There was no difference in overall survival. Toxicities were similar in the two treatment arms. CONCLUSIONS: The paclitaxel-carboplatin-etoposide combination produced a superior overall response rate and time to progression in patients with extensive-stage small cell lung cancer compared with paclitaxel plus topotecan. The platinum compounds continue to be a necessary component of the initial therapy for these patients. PMID- 16249354 TI - Multifocal extranodal non-hodgkin lymphoma: a clinicopathologic study of 37 cases in Greece, a Hellenic Cooperative Oncology Group study. AB - The purpose of this retrospective study was to illustrate the clinicopathological features of patients presenting with multifocal extranodal non-Hodgkin lymphoma (NHL). Among 810 patients with NHL, 37 cases (4.2%) were found to have multiple extranodal involvement (two or more sites). There were 24 men and 13 women, with a median age of 63 years. The majority of these cases (n = 26) had gastric or intestinal (GI) involvement with or without other extranodal sites. Lung along with another extranodal site was relatively common in the present series. Stratification of the 37 cases according to the International Prognostic Index (IPI) showed that 89% of the patients belonged to the high-risk groups. Diffuse large-B-cell lymphoma (DLBCL) accounted for 62%, and mucosa-associated lymphoma tissue (MALT) lymphoma accounted for 27% of all cases. After induction treatment with anthracycline-based regimens, complete remission was achieved in 21 patients (57%), partial remission was achieved in six patients (16%), and seven patients (19%) had no response, while three patients (8%) were nonevaluable. In conclusion, multifocal extranodal NHL is a heterogeneous group of diseases. The majority of them arise at various sites in the GI tract. DLBCL was the most frequent histological subtype followed by MALT lymphoma. Risk group, as defined by the IPI, was predictive of survival. PMID- 16249355 TI - Helping melanoma patients decide whether to choose adjuvant high-dose interferon alpha2b. AB - High-dose interferon-alpha2b is a U.S. Food and Drug Administration-approved adjuvant treatment for stage III melanoma, and yet, because of its limited efficacy and well-known toxicity, it is not universally accepted by patients and oncologists. In this paper, we evaluate the benefits and risks of adjuvant high dose interferon-alpha2b and try to provide a framework to help oncologists guide patients trying to decide whether to undergo adjuvant high-dose interferon therapy. PMID- 16249356 TI - Management of cancer-related anemia in patients with breast or gynecologic cancer: new insights based on results from the European Cancer Anemia Survey. AB - The incidence, prevalence, and treatment of anemia (hemoglobin [Hb] <12 g/dl) in women with breast cancer and gynecologic cancer were evaluated using data from the European Cancer Anemia Survey (ECAS). Adult patients with newly diagnosed treated or untreated disease, persistent/recurrent disease, and disease in remission were enrolled and followed for up to six chemotherapy cycles or six evaluation points within a 6-month period. At enrollment, 30.4% of breast cancer patients and 49.1% of gynecologic cancer patients were anemic. A significant correlation was shown between low Hb level and poor performance status (World Health Organization criteria) at enrollment for both breast cancer and gynecologic cancer patients. In all, 62.4% of breast cancer patients and 81.4% of gynecologic cancer patients were anemic at some time during the survey. The incidence of anemia, determined in a carefully defined population, was 59.8% for breast cancer patients and 74.8% for gynecologic cancer patients. Despite the high prevalence and incidence of anemia, only 26.3% and 42.7% of patients in the respective groups received anemia treatment. In breast cancer patients, the mean Hb trigger was 10 g/dl for epoetin treatment and 8.6 g/dl for transfusion; corresponding values for gynecologic cancer patients were 10.1 g/dl and 9.1 g/dl. Logistic regression analyses in the overall ECAS population identified five factors as significant and suitable predictors of anemia: lower initial Hb, having lung or gynecologic cancer versus gastrointestinal/colorectal cancer, any other cancer versus gastrointestinal/colorectal cancer, treatment with platinum chemotherapy, and being female. The ECAS data highlight the need for greater awareness of the adverse impact of anemia on cancer patients and for optimal anemia management to ensure maximal patient quality of life. PMID- 16249357 TI - The molecular perspective: c-Abl tyrosine kinase. PMID- 16249358 TI - Nimotuzumab: evidence of clinical benefit without rash. PMID- 16249359 TI - Response to "effect of patient exclusion criteria on the efficacy of erythropoiesis-stimulating agents in patients with cancer-related anemia". PMID- 16249361 TI - Interventional pain management: an overview for primary care physicians. AB - Image-guided spine intervention is used primarily for its precise diagnostic capabilities. This article reviews basic principles of the more common image guided diagnostic techniques specifically as they relate to patients with low back pain. It also includes discussion of advanced modes of therapy, including spinal cord stimulation and intrathecal therapy, providing primary care physicians with an understanding of the primary indications for these therapeutic modalities. PMID- 16249362 TI - An osteopathic approach to treating women with chronic pelvic pain. AB - Chronic pelvic pain is a common gynecologic complaint, affecting about 5% of American women. The differential diagnosis is broad, including many medical diseases, surgical indications, musculoskeletal problems, and somatic dysfunction. Women are more affected than men by pelvic pain because their bodies are subject to more changes. These changes include a cyclic hormonal milieu, major alterations in biomechanics during pregnancy, psychosocial stress, and other modifications during childrearing, and more adjustments during menopause. Both medical and surgical approaches to management exist, but integrative modes of therapy address the body-mind-spirit continuum. Osteopathic manipulative treatment is a valuable option for many affected women from childbirth to menopause. PMID- 16249363 TI - Management of osteoarthritic knee pain. AB - Osteoarthritis (OA) is one of the most common forms of arthritis seen in primary care practice. The pain associated with this condition is the chief complaint of most patients, prompting them to seek medical attention. Pain can originate from the synovial membrane, joint capsule, periarticular muscles and ligaments, and periosteum and subchondral bone, among other sources. Osteoarthritis is traditionally thought of as a noninflammatory type of arthritis; however, inflammatory mechanisms can be present. Therefore, the management of osteoarthritic pain involves nonpharmacologic modes of therapy as well as pharmacologic agents. Nonpharmacologic therapeutic modalities include osteopathic manipulative treatment, physical therapy, exercise, use of assistive devices, and weight reduction. Pharmacologic options, categorized as topical, intra-articular, or oral, include acetaminophen, nonsteroidal anti-inflammatory agents, and cyclooxygenase type 2 inhibitors. Patients often benefit from use of a combination of these therapeutic modalities. Although pain relief is a chief motivator for patients with OA to seek medical attention, a secondary benefit of successful treatment is to delay the decreased quality of life associated with osteoarthritic pain. PMID- 16249364 TI - Osteopathic manipulative medicine considerations in patients with chronic pain. AB - Osteopathic manipulative medicine (OMM) incorporates diagnostic and therapeutic strategies that address body unity, homeostatic mechanisms, and structure function interrelationships. In regard to pain, osteopathic physicians take thorough histories guided by palpatory examination to determine the quality, duration, and origin of this condition, how it uniquely affects the individual, and whether segmental, reflex, or triggered pain phenomena coexist. Osteopathic manipulative medicine expands differential diagnoses by considering somatic dysfunction and treatment options by integrating specific aspects of complementary care into state-of-the-art pain management practices. Prescriptions formulated through an OMM algorithm integrate each osteopathic tenet with biopsychosocial and patient education models and medication, rehabilitation, and manual medicine techniques proportionate to individual needs. PMID- 16249365 TI - Headache pain. AB - The headache problem with its debilitation and pain has been noted throughout medical history. It is one of the most common outpatient complaints and affects more than 45 million Americans. The lost days to work and family and the immeasurable suffering of patients can be lessened with the understanding and knowledge of a caring physician. Osteopathic physicians with expertise in holistic and musculoskeletal concepts are particularly well prepared to help. The establishment of an accurate diagnosis through a careful history and physical examination is essential before the physician can develop an effective treatment plan. Treatment can be abortive, prophylactic, or symptomatic, or a combination. Abortive treatment is geared to reverse the headache once begun; prophylactic treatment usually involves the use of daily medications to prevent, decrease frequency, or lessen severity of attacks; and symptomatic treatment is for relief of pain or accompanying symptoms. Most headaches experienced are of the tension type, whereas most debilitating headaches are of the migraine type. Cluster headache, though experienced by a small percentage of sufferers, is especially severe, and is useful in differential diagnosis. PMID- 16249367 TI - Inhibition of leukocyte elastase, polymorphonuclear chemoinvasion, and inflammation-triggered pulmonary fibrosis by a 4-alkyliden-beta-lactam with a galloyl moiety. AB - beta-Lactams, a well known class of antibiotics, have been investigated as inhibitors of the disruptive protease released by inflammatory cells, leukocyte elastase (LE). We have synthesized a new beta-lactam with an N-linked galloyl moiety, the latter identified as strategic in conferring anti-LE properties to some flavonols. This N-galloyl-derivative beta-lactam inhibits the LE activity with a K(i) of 0.7 microM, whereas it exerts weak activity against cathepsin G and protease-3 (IC(50) > 100 microM), and matrix metalloproteinase (MMP)-2 and MMP-9. Without affecting chemotactic response and viability of polymorphonuclear (PMN) leukocytes, the compound efficiently restrains their chemoinvasion (IC(50) of 1-2 microM) blocking the LE-triggered activation of pro-MMP-9, instrumental to extravasation. Daily i.p. injection of compound enhances resolution in a pulmonary inflammation model, significantly reducing consequent fibrosis. These results indicate that the new beta-lactam is a potent anti-inflammatory compound with therapeutic potential. PMID- 16249368 TI - The lymph lipid precursor pool is a key determinant of intestinal lymphatic drug transport. AB - The influence of the size and turnover kinetics of the enterocyte-based lymph lipid precursor pool (LLPP) on intestinal lymphatic drug transport has been examined. Mesenteric lymph duct-cannulated rats were infused intraduodenally with low (2-5 mg/h) or high (20 mg/h) lipid-dose formulations containing 100 microg/h halofantrine (Hf, a model drug) and 1 microCi/h (14)C-oleic acid (OA) (as a marker for lipid transport) until steady-state rates of lipid(dX(L)/dt)(ss) and drug (dD(L)/dt)(ss) transport in lymph were obtained. After 5 h, the infusion was changed to formulations of the same composition but excluding (14)C-OA and Hf, allowing calculation of the first order rate constants describing turnover of lipid (K(X)) and drug (K(D)) from the LLPP into the lymph from the washout kinetics. The mass of lipid (X(LP)) and drug (D(LP)) in the LLPP was also determined. Biliary-lipid output was determined in a separate group of rats that had been infused with the same formulations. The results indicate that after administration of high lipid doses, lymphatic drug transport is dependent on the mass of exogenous lipid available in the LLPP and the rate of lipid pool turnover into the lymph. In contrast, after administration of low lipid doses, biliary derived endogenous lipids are most likely to be the primary drivers of drug incorporation into the LLPP and lymph. Therefore, the LLPP size and composition seem to be major determinants of lymphatic drug transport, and formulation components, which increase lipid pool size, may therefore enhance lymphatic drug transport. PMID- 16249369 TI - A nonthiazolidinedione peroxisome proliferator-activated receptor gamma agonist reverses endothelial dysfunction in diabetic (db/db-/-) mice. AB - We have previously reported that endothelium-dependent relaxation to acetylcholine is impaired in small mesenteric arteries from spontaneously diabetic (db/db) mice. The objective of the present study was to examine the effects of treatment of the db/db and the insulin-resistant ob/ob mice with the PPARgamma agonist 2-(2-(4-phenoxy-2-propylphenoxy)ethyl)indole-5-acetic acid (COOH). In the db/db model, an 8-week treatment with COOH (30 mg/kg/day) reduced plasma glucose from 48.0 +/- 2.5 (untreated) to 12.6 +/- 1.1 mM. In contrast, plasma glucose was not elevated in untreated ob/ob mice. Relaxation of small mesenteric arteries mediated by acetylcholine was impaired in the untreated db/db diabetic mice (51.7 +/- 7.4% maximal relaxation, n = 6) but not in the ob/ob mice (70.8 +/- 8.6% maximal relaxation, n = 3). This impairment was reversed with COOH treatment (86.9 +/- 0.4% maximal relaxation, n = 5). Malondialdehyde was elevated in plasma from diabetic db/db mice (13.9 +/- 1.1 versus 12.0 +/- 0.7 micromol/ml); however, when normalized to total cholesterol, no significant differences in the ratio of lipid peroxidation in plasma were identified. Western blot analysis and quantitative polymerase chain reaction for eNOS was performed on the isolated mesenteric vessels and revealed no differences in the relative levels of eNOS expression in diabetic and control animals; in addition, treatment with COOH had no significant effect on eNOS levels in either group. In summary, endothelial dysfunction and hyperglycemia were completely normalized in COOH treated db/db mice. In contrast, nonhyperglycemic ob/ob mice exhibited normal vasodilatory responses to acetylcholine and, consequently, COOH treatment had no effect on endothelial function. PMID- 16249370 TI - Apolipoprotein E-derived peptides block alpha7 neuronal nicotinic acetylcholine receptors expressed in xenopus oocytes. AB - For decades, the pathology of Alzheimer's disease has been associated with dysfunction of cholinergic signaling; however, the cellular mechanisms by which nicotinic acetylcholine receptor (nAChR) function is impaired in Alzheimer's disease are as yet unknown. The most significant genetic risk factor for the development of Alzheimer's disease is inheritance of the epsilon4 allele of apolipoprotein E (apoE). Recent data have demonstrated the ability of apoE derived peptides to inhibit nAChRs in rat hippocampus. In the current study, the functional interaction between nAChRs and apoE-derived peptides was investigated in Xenopus oocytes expressing selected nAChRs. Both a 17-amino acid peptide fragment, apoE(133-149), and an eight-amino acid peptide, apoE(141-148), were able to maximally block acetylcholine (ACh)-mediated peak current responses for homomeric alpha7 nAChRs. ApoE peptide inhibition was dose-dependent and voltage- and activity-independent. The current findings suggest that apoE peptides are noncompetitive for acetylcholine and do not block functional alpha-bungarotoxin binding. ApoE peptides had a significantly decreased ability to inhibit ACh mediated peak current responses for alpha4beta2 and alpha2beta2 nAChRs. Amino acid substitutions in the apoE peptide sequence suggest that the arginines are critical for peptide blockade of the alpha7 nAChR. The current data suggest that apoE fragments can disrupt nAChR signaling through a direct blockade of alpha7 nAChRs. These results may be useful in elucidating the mechanisms underlying memory loss and cognitive decline seen in Alzheimer's disease as well as aid in the development of novel therapeutics using apoE-derived peptides. PMID- 16249371 TI - Sex differences in the mRNA, protein, and functional expression of organic anion transporter (Oat) 1, Oat3, and organic cation transporter (Oct) 2 in rabbit renal proximal tubules. AB - Sex differences in transport of the organic anion (OA) substrate p-aminohippurate (PAH) and the organic cation (OC) substrate tetraethylammonium (TEA) have been recognized for some time. In the rat kidney, androgens up-regulate and estrogens down-regulate PAH and TEA transport, which correlate with similar changes in mRNA and protein expression for the renal basolateral membrane transporters organic anion transporter (Oat) 1 and organic cation transporter (Oct) 2. However, these sex differences are not readily demonstrated in other species. The present study characterizes the kinetics of basolateral membrane PAH, estrone sulfate (ES), and TEA uptake in renal proximal tubule (RPT) suspensions isolated from female and male rabbits to compare functional expression of transport with mRNA and protein expression for rbOat1, rbOat3, and rbOct2. Although rbOat1-rbOat3 mRNA expression exhibited developmental differences, no sex differences in mRNA levels were observed. Oat1 and Oat3 protein expression in RPT suspensions also was similar between adult female and male rabbits. In contrast, rbOct1 and rbOct2 mRNA levels did not show developmental differences, but rbOct2 mRNA expression was greater in adult male than female rabbits. However, the sex difference in rbOct2 mRNA level did not translate to rbOct2 protein expression. Importantly, functional expression of Oat1, Oat3, and Oct2 transport as measured by kinetics (J(max) and K(t)) of PAH, ES, and TEA uptake was similar between adult male and female rabbits, and correlated with rbOat1, rbOat3, and rbOct2 protein expression. Thus, unlike rodents, rabbit renal OA and OC transport does not exhibit sex differences, pointing to the need for caution in extrapolating transport-related sex differences between species. PMID- 16249372 TI - Presynaptic alpha1 adrenergic receptors differentially regulate synaptic glutamate and GABA release to hypothalamic presympathetic neurons. AB - The hypothalamic paraventricular nucleus (PVN) neurons that project to the spinal intermediolateral cell column and brainstem are important for the control of sympathetic outflow. Stimulation of alpha(1) adrenergic receptors in the PVN increases sympathetic outflow, but the cellular mechanisms remain unclear. In this study, we determined the role of alpha(1) adrenergic receptors in the regulation of glutamatergic and GABAergic synaptic inputs to spinally projecting PVN neurons. Whole-cell and cell-attached patch-clamp recordings were performed on retrogradely labeled PVN-spinal neurons in rat brain slices. Bath application of 10 to 100 microM phenylephrine, an alpha(1) adrenergic receptor agonist, significantly increased the frequency of spontaneous excitatory postsynaptic currents in a concentration-dependent manner. This effect was blocked by the alpha (1)adrenergic receptor antagonists prazosin or corynanthine. Phenylephrine also significantly increased the frequency of miniature excitatory postsynaptic currents (mEPSCs) but not the amplitude and decay constant of mEPSCs. Furthermore, activation of alpha(1) adrenergic receptors with phenylephrine or cirazoline significantly decreased the frequency of spontaneous inhibitory postsynaptic currents and miniature inhibitory postsynaptic currents, and this effect also was blocked by corynanthine. In addition, 50 microM phenylephrine significantly increased the firing rate of 13 labeled PVN neurons from 3.16 +/- 0.42 to 5.83 +/- 0.65 Hz. However, phenylephrine failed to increase the firing of most labeled PVN neurons in the presence of GABA(A) and ionotropic glutamate receptor antagonists. Thus, these data suggest that activation of alpha (1)adrenergic receptors increases the excitability of PVN presympathetic neurons primarily through augmentation of glutamatergic tone and attenuation of GABAergic inputs. PMID- 16249373 TI - delta opioid receptors stimulate Akt-dependent phosphorylation of c-jun in T cells. AB - Activation of naive T cells markedly up-regulates the expression of delta opioid receptors (DORs). These receptors are bound by DOR peptides released by T cells, modulating T cell functions such as interleukin-2 production, cellular proliferation, and chemotaxis. Previous studies have shown that DOR agonists [e.g., [D-Ala(2)-D-Leu(5)]-enkephalin (DADLE)] modulate T cell antigen receptor signaling through mitogen-activated protein kinases (MAPKs; i.e., extracellular signal-regulated kinases 1 and 2) and that DORs directly induce phosphorylation of activating transcription factor-2 (implicated in cytokine gene transcription) and its association with the MAPK c-jun1 NH(2)-terminal kinase (JNK). Such observations suggest that DORs may induce the phosphorylation of c-jun. These experiments were performed to test this hypothesis and determine the potential roles of phosphoinositide 3-kinase (PI3K) and Akt (protein kinase B). DADLE (10( 10) to 10(-6) M) dose-dependently induced c-jun phosphorylation. This was blocked by pertussis toxin and the DOR-specific antagonist naltindole. Fluorescence flow cytometry showed that DADLE significantly stimulated c-jun phosphorylation by T cells. DADLE stimulated phosphorylation of membrane-associated Akt; wortmannin and LY294002 ([2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one]), specific inhibitors of PI3K, abolished the DADLE-induced phosphorylation of c-jun. Finally, inhibitors of Akt and JNK blocked DADLE-induced phosphorylation of c jun. Thus, activated DORs directly stimulate c-jun phosphorylation through a PI3K dependent pathway in T cells, apparently involving Akt. This implies that DORs activate JNK through a novel pathway dependent on PI3K and Akt, thereby regulating the function of activator protein-1 transcription complexes containing c-jun and other transcription partners. PMID- 16249374 TI - Hydroxylated xestospongins block inositol-1,4,5-trisphosphate-induced Ca2+ release and sensitize Ca2+-induced Ca2+ release mediated by ryanodine receptors. AB - Inositol-1,4,5-trisphosphate receptors (IP(3)Rs) and ryanodine receptors (RyRs) often coexist within the endoplasmic/sarcoplasmic reticulum (ER/SR) membrane and coordinate precise spatial and temporal coding of Ca(2+) signals in most animal cells. Xestospongin C (XeC) was shown to selectively block IP(3)-induced Ca(2+) release and IP(3)R-mediated signaling (Gafni et al., 1997). We have further studied the specificity of xestospongin structures possessing ring hydroxyl (-OH) substituents toward IP(3)R, RyR, and ER/SR Ca(2+)-ATPase (SERCA) activities. XeC potently inhibits IP(3)R, weakly inhibits RyR1, and lacks activity toward SERCA1 and SERCA2. XeD (9-OH XeC), 7-OH-XeA, and araguspongin C isolated from the marine sponge Xestospongia species also inhibit IP(3)-mediated Ca(2+) release and lack activity toward SERCA. However, these hydroxylated derivatives possess a unique activity in that they enhance Ca(2+)-induced Ca(2+) release from SR vesicles by a mechanism involving the sensitization of RyR1 channels within the same concentration range needed to block IP(3)-induced Ca(2+) release. These results show that xestospongins and related structures lack direct SERCA inhibitory activity, as suggested by some previous studies. A new finding is that XeD and related structures possessing a hydroxylated oxaquinolizidine ring are IP(3)R blockers that also enhance Ca(2+)-induced Ca(2+) release mediated by RyRs. In intact cells, the actions of XeD are blocked by ryanodine pretreatment and do not interfere with thapsigargin-mediated Ca(2+) mobilization stemming from SERCA block. Hydroxylated bis-oxaquinolizadine derivatives isolated from Xestospongia species are novel bifunctional reagents that may be useful in ascertaining how IP(3)Rs and RyRs contribute to cell signaling. PMID- 16249375 TI - Anti-Epo receptor antibodies do not predict Epo receptor expression. AB - Investigators using anti-EpoR antibodies for immunoblotting and immunostaining have reported erythropoietin receptor (EpoR) expression in nonhematopoietic tissues including human tumors. However, these antibodies detected proteins of 66 to 78 kDa, significantly larger than the predicted molecular weight of EpoR (56 57 kDa). We investigated the specificity of these antibodies and showed that they all detected non-EpoR proteins. C-20 detected 3 proteins in tumor cell lines (35, 66, and 100 kDa). Sequences obtained from preparative gels had similarity to the C-20-immunizing peptide. The 66-kDa protein was a heat shock protein (HSP70) to which antibody binding was abrogated in peptide competition experiments. Antibody M-20 readily identified a 59-kDa EpoR protein. However, neither M-20 nor C-20 was suitable for detection of EpoR using immunohistochemical methods. We concluded that these antibodies have limited utility for detecting EpoR. Thus, reports of EpoR expression in tumor cells using these antibodies should be viewed with caution. PMID- 16249376 TI - Human immunoglobulin inhibits liver transduction by AAV vectors at low AAV2 neutralizing titers in SCID mice. AB - Long-term cures of hemophilia B have been achieved using AAV2 delivering the factor IX gene to the liver of adeno-associated virus (AAV)-naive hemophilic animals. However, the clinical success of this approach requires overcoming pre existing AAV neutralizing antibodies prevalent in humans. To better define the inhibition of neutralizing antibodies on AAV2-mediated liver transduction, we developed an in vivo passive immunity model. SCID mice were first reconstituted to a defined neutralizing titer with pooled plasma-derived human immunoglobulin. AAV2-FIX vectors then were administered to the liver, and the transduction efficiency was measured by plasma FIX levels. Unexpectedly, AAV2 neutralizing titers lower than 1:10 were sufficient to neutralize 4 to 20 x 10(12) vg/kg of AAV2 vectors in vivo, a capacity that was underestimated by in vitro neutralizing assays. We also evaluated strategies to evade neutralization, including the use of alternative delivery routes, infusion parameters, empty capsids, and alternative AAV serotypes 6 and 8. The results indicate that low AAV2 neutralizing titers can be inhibitory to the tested human and primate AAV vectors delivered into the circulatory system. Therefore, novel nonprimate AAV vectors or compartmentalized delivery may offer more consistent therapeutic effects in the presence of pre-existing AAV neutralizing antibodies. PMID- 16249378 TI - BCL6 is regulated by p53 through a response element frequently disrupted in B cell non-Hodgkin lymphoma. AB - The BCL6 transcriptional repressor mediates survival, proliferation, and differentiation blockade of B cells during the germinal-center reaction and is frequently misregulated in B-cell non-Hodgkin lymphoma (BNHL). The p53 tumor suppressor gene is central to tumorigenesis. Microarray analysis identified BCL6 as a primary target of p53. The BCL6 intron 1 contains a region in which 3 types of genetic alterations are frequent in BNHL: chromosomal translocations, point mutations, and internal deletions. We therefore defined it as TMDR (translocations, mutations, and deletions region). The BCL6 gene contains a p53 response element (p53RE) residing within the TMDR. This p53RE contains a motif known to be preferentially targeted by somatic hypermutation. This p53RE is evolutionarily conserved only in primates. The p53 protein binds to this RE in vitro and in vivo. Reporter assays revealed that the BCL6 p53RE can confer p53 dependent transcriptional activation. BCL6 mRNA and protein levels increased after chemotherapy/radiotherapy in human but not in murine tissues. The increase in BCL6 mRNA levels was attenuated by the p53 inhibitor PFT-alpha. Thus, we define the BCL6 gene as a new p53 target, regulated through a RE frequently disrupted in BNHL. PMID- 16249379 TI - Frequency of telomerase-specific CD8+ T lymphocytes in patients with cancer. AB - Telomerase is considered a universal tumor-associated antigen (TAA) due to its high rate of expression by cancers (approximately 90%), and clinical trials are in progress to test the immunotherapeutical efficacy of antitelomerase immunization in patients with cancer. However, the data concerning frequency and functional activity of telomerase-specific cytotoxic T lymphocytes (CTLs) in patients with cancer are few and conflicting, although their knowledge would be mandatory to predict the efficacy of telomerase-specific immunotherapy in selected patients. We performed this study to analyze frequency and cytolytic function of circulating CD8+ T lymphocytes specific for the p540 telomerase peptide in a series of human leukocyte antigen (HLA)-A2+ cancer patients. The results show that most patients with cancer have circulating telomerase-specific CD8+ T lymphocytes, but a high frequency of telomerase-specific CTLs are present only in a fraction of them. Furthermore, CTL lines able to kill telomerase positive tumor cells, including autologous cancer cells, can be expanded ex vivo from some, but not all, patients with cancer. In conclusion, the results of the study support the development of clinical protocols using telomerase peptides as an immunizing agent. However, they underline the necessity to study single patients immunologically before undergoing vaccination, to select the patients adequately, and to eventually adapt the immunization schedule to the patient's immunologic status. PMID- 16249377 TI - Nitric oxide and cGMP protein kinase (cGK) regulate dendritic-cell migration toward the lymph-node-directing chemokine CCL19. AB - Dendritic-cell (DC) migration to secondary lymphoid organs is crucial for the initiation of adaptive immune responses. Although LPS up-regulates CCR7 on DCs, a second signal is required to enable them to migrate toward the chemokine CCL19 (MIP-3beta). We found that the nitric oxide (NO) donor NOR4 provides a signal allowing LPS-stimulated DCs to migrate toward CCL19. NO affects DC migration through both the initial activation of the cGMP/cGMP kinase (cGMP/cGK) pathway and a long-term effect that reduced cGK activity via negative feedback. Indeed, migration of DCs toward CCL19, unlike migration toward CXCL12 (SDF-1alpha), required inhibition of cGK. LPS increased both cGK expression and cGK activity as measured by phosphorylation of the key cGK target vasodilator-stimulated phosphoprotein (VASP). Because cGK phosphorylation of VASP can disrupt focal adhesions and inhibit cell migration, LPS-induced VASP phosphorylation may prevent DCs from migrating without a second signal. Long-term NOR4 treatment inhibited the increase in cGK-dependent VASP phosphorylation, releasing this brake so that DCs can migrate. NO has been implicated in the regulation of autoimmunity through its effect on T cells. Our results suggest that NO regulation of DC migration and cytokine production may contribute to the protective effects of NO in autoimmune disorders. PMID- 16249380 TI - G-CSF-treated granulocytes inhibit acute graft-versus-host disease. AB - It has been shown that in vivo and in vitro treatment with G-CSF induces the generation of low-density granulocytes (LDGs), which copurify with PBMCs and inhibit IFN-gamma production by human T cells. These results prompted us to postulate an immunomodulatory role for LDGs in acute graft-versus-host disease (aGVHD). Here it is shown that in the mouse experimental model, in vivo and in vitro G-CSF treatment generates LDGs capable of inhibiting 80% of T-cell IFN gamma production. To assess the role of these LDGs in aGVHD, lethally irradiated (C57BL/6 x BALB/c) F1 hosts were reconstituted with T cell-depleted bone marrow cells plus nylon wool-purified spleen cells from G-CSF-treated (G-NWS) or nontreated (NWS) C57BL/6 donors. Recipients of G-NWS had a 75% survival rate in contrast to a rate of 25% in the NWS recipients. The protective effect was completely abolished, and the mortality rate was 100% if donor-cell infusion was treated with anti-Gr1. Moreover, if LDGs were infused with NWS, full protection of aGVHD was observed, and no signs of disease were evidenced by mortality rate, weight loss, or histopathology of target organs. These results revealed the unexpected immunosuppressive capacity of G-CSF based on the generation of LDGs, leading to the possibility of using these cells as inhibitors of aGVHD. PMID- 16249381 TI - Primitive human hematopoietic cells give rise to differentially specified daughter cells upon their initial cell division. AB - It is often predicted that stem cells divide asymmetrically, creating a daughter cell that maintains the stem-cell capacity, and 1 daughter cell committed to differentiation. While asymmetric stem-cell divisions have been proven to occur in model organisms (eg, in Drosophila), it remains illusive whether primitive hematopoietic cells in mammals actually can divide asymmetrically. In our experiments we have challenged this question and analyzed the developmental capacity of separated offspring of primitive human hematopoietic cells at a single-cell level. We show for the first time that the vast majority of the most primitive, in vitro-detectable human hematopoietic cells give rise to daughter cells adopting different cell fates; 1 inheriting the developmental capacity of the mother cell, and 1 becoming more specified. In contrast, approximately half of the committed progenitor cells studied gave rise to daughter cells, both of which adopted the cell fate of their mother. Although our data are compatible with the model of asymmetric cell division, other mechanisms of cell fate specification are discussed. In addition, we describe a novel human hematopoietic progenitor cell that has the capacity to form natural killer (NK) cells as well as macrophages, but not cells of other myeloid lineages. PMID- 16249383 TI - VEGFR-1 (FLT-1) activation modulates acute lymphoblastic leukemia localization and survival within the bone marrow, determining the onset of extramedullary disease. AB - The presence of persistent circulating leukemia cells, or engrafted into extramedullary tissues, is a bad prognostic factor for patients with acute leukemia. However, little is known about the mechanisms that regulate the exit of leukemia cells from the bone marrow (BM) microenvironment. We reveal that vascular endothelial growth factor receptor 1 (FLT-1) modulates acute leukemia distribution within the BM, along VEGF and PlGF gradients, regulating leukemia survival and exit into the peripheral circulation. FLT-1 activation on acute lymphoblastic leukemia (ALL) cells results in cell migration and proliferation in vitro, whereas in vivo FLT-1-overexpressing cells accumulate in the BM epiphysis of nonobese diabetic-severe combined immunodeficient (NOD-SCID) recipients and are detected in circulation 2 weeks after inoculation. In turn, FLT-1 neutralization affects leukemia localization (now in the BM diaphysis), increases leukemia apoptosis, and impedes the exit of ALL cells, prolonging the survival of inoculated mice. We demonstrate further that FLT-1-induced cell migration involves actin polymerization and lipid raft formation. Taken together, we show that FLT-1 regulates the BM localization of ALL cells, determining their survival and exit into the circulation and ultimately the survival of inoculated recipients. FLT-1 targeting on subsets of acute leukemias may delay the onset of extramedullary disease, which may be advantageous in combinatorial therapeutic settings. PMID- 16249382 TI - An amphipathic motif at the transmembrane-cytoplasmic junction prevents autonomous activation of the thrombopoietin receptor. AB - Ligand binding to the thrombopoietin receptor (TpoR) is thought to impose a dimeric receptor conformation(s) leading to hematopoietic stem cell renewal, megakaryocyte differentiation, and platelet formation. Unlike other cytokine receptors, such as the erythropoietin receptor, TpoR contains an amphipathic KWQFP motif at the junction between the transmembrane (TM) and cytoplasmic domains. We show here that a mutant TpoR (delta5TpoR), where this sequence was deleted, is constitutively active. In the absence of ligand, delta5TpoR activates Jak2, Tyk2, STAT5, and mitogen-activated protein (MAP) kinase, but does not appear to induce STAT3 phosphorylation. Delta5TpoR induces hematopoietic myeloid differentiation in the absence of Tpo. In the presence of Tpo, the delta5TpoR mutant appears to enhance erythroid differentiation when compared with the Tpo activated wild-type TpoR. Strikingly, individual substitution of K507 or W508 to alanine also induces constitutive TpoR activation, indicating that the K and W residues within the amphipathic KWQFP motif are crucial for maintaining the unliganded receptor inactive. These residues may be targets for activating mutations in humans. Such a motif may exist in other receptors to prevent ligand independent activation and to allow signaling via multiple flexible interfaces. PMID- 16249384 TI - Coordination of intrinsic, extrinsic, and endoplasmic reticulum-mediated apoptosis by imatinib mesylate combined with arsenic trioxide in chronic myeloid leukemia. AB - A treatment strategy that combines arsenic trioxide (ATO) with the tyrosine kinase inhibitor imatinib mesylate (STI571, Gleevec) appears to induce markedly more cell apoptosis than imatinib mesylate alone in chronic myeloid leukemia (CML). To understand the mechanisms underlying the synergistic/additive action of these agents, we applied cDNA microarrays, component plane presentation integrated self-organizing map (CPP-SOM), and methods of protein biochemistry to study cell apoptosis induced by imatinib mesylate, ATO, and the combination of the 2 agents in the CML cell line K562. Numerous features with temporospatial relationships were revealed, indicating the coordinated regulation of molecular networks from various aspects of proapoptotic and apoptotic activities in CML. Imatinib mesylate appears to induce mainly the intrinsic pathway of cell apoptosis, whereas ATO induces the endoplasmic reticulum (ER) stress-mediated pathway of cell apoptosis, and the combination of the 2 agents seems to more effectively induce the intrinsic, extrinsic, and ER stress-mediated pathways of cell apoptosis, which results in a more effective and efficient induction of programmed cell death in K562 cells. This finding appears to be supported also by data derived from bone marrow cells of 2 patients with CML, one in chronic phase and the other in blast-crisis phase of the disease. PMID- 16249385 TI - Differential gene expression, GATA1 target genes, and the chemotherapy sensitivity of Down syndrome megakaryocytic leukemia. AB - Children with Down syndrome (DS) with acute megakaryocytic leukemia (AMkL) have very high survival rates compared with non-DS AMkL patients. Somatic mutations identified in the X-linked transcription factor gene, GATA1, in essentially all DS AMkL cases result in the synthesis of a shorter (40 kDa) protein (GATA1s) with altered transactivation activity and may lead to altered expression of GATA1 target genes. Using the Affymetrix U133A microarray chip, we identified 551 differentially expressed genes between DS and non-DS AMkL samples. Transcripts for the bone marrow stromal-cell antigen 2 (BST2) gene, encoding a transmembrane glycoprotein potentially involved in interactions between leukemia cells and bone marrow stromal cells, were 7.3-fold higher (validated by real-time polymerase chain reaction) in the non-DS compared with the DS group. Additional studies confirmed GATA1 protein binding and transactivation of the BST2 promoter; however, stimulation of BST2 promoter activity by GATA1s was substantially reduced compared with the full-length GATA1. CMK sublines, transfected with the BST2 cDNA and incubated with HS-5 bone marrow stromal cells, exhibited up to 1.7 fold reduced cytosine arabinoside (ara-C)-induced apoptosis, compared with mock transfected cells. Our results demonstrate that genes that account for differences in survival between DS and non-DS AMkL cases may be identified by microarray analysis and that differential gene expression may reflect relative transactivation capacities of the GATA1s and full-length GATA1 proteins. PMID- 16249387 TI - LIME acts as a transmembrane adapter mediating BCR-dependent B-cell activation. AB - Assembly of a signaling complex around the transmembrane adapter LAT is essential for the transmission of T-cell receptor (TCR)-mediated signaling. However, a LAT like molecule responsible for the initial activation events in B-cell receptor (BCR) signaling has not yet been identified. Here, we show that LIME is a transmembrane adaptor required for BCR-mediated B-cell activation. LIME was found to be expressed in mouse splenic B cells. Upon BCR cross-linking, LIME was tyrosine phosphorylated by Lyn and associated with Lyn, Grb2, PLC-gamma2, and PI3K. Reduction of LIME expression by the introduction of siRNA resulted in the disruption of BCR-mediated activation of MAPK, calcium flux, NF-AT, PI3K, and NF kappaB. Taken together, these results establish that LIME is an essential transmembrane adaptor linking BCR ligation to the downstream signaling events that lead to B-cell activation. PMID- 16249388 TI - Hydrogen peroxide inhibits IL-12 p40 induction in macrophages by inhibiting c-rel translocation to the nucleus through activation of calmodulin protein. AB - Although the antimicrobial activity of reactive oxygen species (ROSs) is well defined, the role of ROSs in regulating the immune response of the body is not well understood. We now provide evidence that hydrogen peroxide (H2O2), a major component of ROSs, inhibits interleukin-12 (IL-12) p40 and IL-12 p70 induction in murine macrophages and catalase pretreatment prevents H2O2-mediated down regulation of IL-12. Endogenous accumulation of H2O2/ROSs in macrophages treated with alloxan resulted in IL-12 p40 inhibition. Although nuclear expression of both p50 and p65 NF-kappaB increased on H2O2 exposure, nuclear c-rel level was inhibited. Overexpression of c-rel restored IL-12 p40 on stimulation with lipopolysaccharide plus IFN-gamma during H2O2 treatment. H2O2 did not inhibit c rel induction in cytosol; however, it prevented the transport of c-rel from cytosol to the nucleus. H2O2 activated calmodulin (CaM) protein in the cytosol, which subsequently sequestered c-rel in the cytosol preventing its transport to the nucleus. The CaM inhibitor trifIuoperazine increased both nuclear c-rel and IL-12 p40 levels in H2O2-treated macrophages, emphasizing a role of CaM in these processes. H2O2/ROSs thus down-regulate IL-12 induction in macrophages by a novel pathway inhibiting c-rel translocation to the nucleus through activation of CaM protein. PMID- 16249386 TI - BCR-ABL nuclear entrapment kills human CML cells: ex vivo study on 35 patients with the combination of imatinib mesylate and leptomycin B. AB - The BCR-ABL oncoprotein of chronic myelogenous leukemia (CML) localizes to the cell cytoplasm, where it activates proliferative and antiapoptotic signaling pathways. We previously reported that the combination of the ABL kinase inhibitor imatinib mesylate (IM) and the nuclear export inhibitor leptomycin B (LMB) traps BCR-ABL inside the nucleus, triggering the death of the leukemic cells. To evaluate the efficacy of the combination of IM and LMB on human cells we collected CD34-positive cells from 6 healthy donors and myeloid progenitors from 35 patients with CML. The sequential addition of IM and LMB generated the strongest reduction in the proliferative potential of the leukemic cells, with limited toxicity to normal myeloid precursors. Furthermore, nested reverse transcriptase-polymerase chain reaction (RT-PCR) analysis on colonies representative of each experimental condition demonstrated that the combination of IM and LMB was the most effective regimen in reducing the number of BCR-ABL positive colonies. The efficacy of the 2-drug association was independent of the clinical characteristics of the patients. Our results indicate that strategies aimed at the nuclear entrapment of BCR-ABL efficiently kill human leukemic cells, suggesting that the clinical development of this approach could be of significant therapeutic value for newly diagnosed and IM-resistant CML patients. PMID- 16249389 TI - Tumor rejection by the poliovirus receptor family ligands of the DNAM-1 (CD226) receptor. AB - The poliovirus receptor CD155 and its family member CD112 (nectin-2) are the ligands for the activating cell-surface receptor DNAM-1 on CD8+ T cells and natural killer (NK) cells. Here, we demonstrate that, whereas the RMA tumor grew in syngeneic mice, DNAM-1 ligand-transduced RMA was rejected, in which CD8+ T cells and NK cells played an essential role. Importantly, CD8+ memory cytotoxic T cells to parental RMA were generated in these mice. We found that DNAM-1 was also expressed on CD8alpha+, rather than CD8alpha-, dendritic cells (DCs). Cross linking DNAM-1 induced maturation of CD8alpha+ DCs. Antigen presentation by these stimulated DCs drove Th1 cells. Moreover, the rejection of DNAM-1 ligand transduced RMA was canceled in CD4+ T-cell-depleted and major histocompatibility complex class II-deficient mice. Taken together, these results suggest that DNAM 1 ligands stimulate innate immunity by CD8alpha+ DCs as well as NK cells, which efficiently prime cell-mediated tumor-specific immunity. PMID- 16249393 TI - Prisonization and sexual offenders: a compounded problem. PMID- 16249390 TI - Regulation of ITAM-positive receptors: role of IL-12 and IL-18. AB - Our previous studies have identified mechanisms by which cytokine production, blocked by Ly49G2 receptor cross-linking, can be overridden. In this study we analyzed the regulation of other ITAM-positive receptor signaling on NK, NKT, and T cells and characterized the biochemical pathways involved in this signaling. Our studies demonstrate that cross-linking of NKG2D and NK1.1 results in a synergistic NK IFN-gamma response when combined with IL-12 or IL-18. Examination of NKT- and T-cell responses demonstrated that cross-linking of NKG2D and CD3 resulted in potent synergy when combined with IL-12 and, to a lesser degree, with IL-18. We have now found that both the p38 MAP kinase and the ERK-dependent signal transduction pathways are required for the synergistic response. Further mechanistic examination of the synergy indicated a potent up-regulation of total IFN-gamma mRNA in the nuclear and the cytoplasmic compartment, but mRNA half-life was not affected. Fifteen minutes of IL-12 pretreatment was sufficient to result in maximal synergistic activation, indicating that the response of the cells to the IL-12 signal was rapid and immediate. Thus, our data demonstrate that multiple convergent signals maximize the innate immune response by triggering complementary biochemical signaling pathways. PMID- 16249391 TI - Expansion of HIV-specific CD4+ and CD8+ T cells by dendritic cells transfected with mRNA encoding cytoplasm- or lysosome-targeted Nef. AB - Transfection with synthetic mRNA is a safe and efficient method of delivering antigens to dendritic cells for immunotherapy. Targeting antigens to the lysosome can sometimes enhance the CD4+ T-cell response. We transfected antigen-presenting cells (APCs) with mRNA encoding Gag-p24 and cytoplasmic, lysosomal, and secreted forms of Nef. Antigen-specific cytotoxic T cells were able to lyse the majority of transfected targets, indicating that transfection was efficient. Transfection of APCs with a Nef construct bearing lysosomal targeting signals produced rapid and prolonged antigen presentation to CD4+ and CD8+ T cells. Polyclonal CD4+ and CD8+ T-cell lines recognizing multiple distinct epitopes were expanded by coculture of transfected dendritic cells with peripheral blood mononuclear cells from viremic and aviremic HIV-infected subjects. Importantly, lysosome-targeted antigen drove a significantly greater expansion of Nef-specific CD4+ T cells than cytoplasmic antigen. The frequency of recognition of CD8 but not CD4 epitopes by mRNA-expanded T cells was inversely proportional to sequence entropy and was similar to ex vivo responses from a large chronic cohort. Thus human dendritic cells transfected with mRNA encoding lysosome-targeted HIV antigen can expand a broad, polyclonal repertoire of antiviral T cells, offering a promising approach to HIV immunotherapy. PMID- 16249394 TI - Learning to pass: sex offenders' strategies for establishing a viable identity in the prison general population. AB - This article endeavors to illustrate the realities of prison life for sex offenders and the means by which they attempt to establish viable identities and acquire a survivable niche in the prison general population, particularly when established identities and protective niches are put at risk by entry into a sex offender treatment program. Qualitative data was collected by repeatedly interviewing a cohort of sex offenders for 6 months as they completed a basic sex offender treatment program. The findings indicate a need to include consideration of treatment context in understanding the limits of treatment gain in prison based programs. PMID- 16249395 TI - Psychosocial adjustment and psychopathology of men sexually abused during childhood. AB - The purpose of this study was to evaluate psychosocial adjustment and psychopathology among men sexually abused during their childhood and arrive at a better understanding of the reasons for which, following such sexual experiences, some adult males manifest greater distress than others. In total, 64 participants were interviewed and completed eight questionnaires covering various aspects of childhood and adult life. Results suggest the existence of three types of sexual abuse scenarios. Participants in each of these three groups presented with an adult-specific adjustment. However, the methodology and multivariate analyses used in this study suggest that some of these problems were not only associated with the sexual contacts but also with other childhood experiences and with the participant's cognitive, affective, and psychological resources. PMID- 16249392 TI - Immunotherapy of high-risk acute leukemia with a recipient (autologous) vaccine expressing transgenic human CD40L and IL-2 after chemotherapy and allogeneic stem cell transplantation. AB - CD40L generates immune responses in leukemia-bearing mice, an effect that is potentiated by IL-2. We studied the feasibility, safety, and immunologic efficacy of an IL-2- and CD40L-expressing recipient-derived tumor vaccine consisting of leukemic blasts admixed with skin fibroblasts transduced with adenoviral vectors encoding human IL-2 (hIL-2) and hCD40L. Ten patients (including 7 children) with high-risk acute myeloid (n = 4) or lymphoblastic (n = 6) leukemia in cytologic remission (after allogeneic stem cell transplantation [n = 9] or chemotherapy alone [n = 1]) received up to 6 subcutaneous injections of the IL-2/CD40L vaccine. None of the patients were receiving immunosuppressive drugs. No severe adverse reactions were noted. Immunization produced a 10- to 890-fold increase in the frequencies of major histocompatibility complex (MHC)-restricted T cells reactive against recipient-derived blasts. These leukemia-reactive T cells included both T-cytotoxic/T-helper 1 (Th1) and Th2 subclasses, as determined from their production of granzyme B, interferon-gamma, and interleukin-5. Two patients produced systemic IgG antibodies that bound to their blasts. Eight patients remained disease free for 27 to 62 months after treatment (5-year overall survival, 90%). Thus, even in heavily treated patients, including recipients of allogeneic stem cell transplants, recipient-derived antileukemia vaccines can induce immune responses reactive against leukemic blasts. This approach may be worthy of further study, particularly in patients with a high risk of relapse. PMID- 16249396 TI - Aggression and anxiety in rapists and child molesters. AB - This study expands the empirical and theoretical understanding of the distinction between those who perpetrate sexual assaults against children (child molesters) and those who perpetrate them against adults (rapists). Two questionnaires were completed by 88 incarcerated, male sexual offenders (45 child molesters and 43 rapists): the Spielberger state trait anxiety inventory and the Buss-Durkee hostility inventory. It was found that the rapists' level of aggression was significantly higher than that of the child molesters. No significant differences in anxiety levels were found between the two groups. The results are discussed in terms of their relevance to theory and clinical practice. PMID- 16249397 TI - Cognitive processes in criminal profile construction: a preliminary study. AB - This study undertook an empirically based examination of the cognitive processes associated with the accurate construction of a criminal psychological profile. This was accomplished by comparing the abilities of profilers and nonprofilers in two simulated profiling exercises that measured both profile accuracy and an individual's performance on various tests of memory and comprehension related to the case materials presented in each exercise. The results of these experiments suggest that an incremental relationship exists between comprehension of the case materials and accuracy of the profiles generated. In addition, the findings provide some tentative indications that the comprehension of case material in a narrative (i.e., written) format is an integral cognitive function to proficient profiling. PMID- 16249398 TI - Exploring the type-of-crime hypothesis, religiosity, and spirituality in an adult male prison population. AB - This study examined the association between measures of spirituality and religiosity and characteristics of current criminal conviction in a male prison population. Spirituality was operationalized as spiritual well-being and existential well-being. Religiosity was operationalized as frequency of religious service attendance, whether an individual considered himself to be religious, and how strongly an individual believed his religious beliefs influenced his behavior. Inmates whose convictions were property related reported greater spiritual wellbeing, were more likely to consider themselves religious, and to say that religious beliefs influenced their behavior than inmates whose crimes were not property related. Inmates whose convictions were drug related were less likely to consider themselves religious, and inmates whose conviction involved violence were more likely to consider themselves religious but less likely to endorse statements that religious beliefs influenced their behavior. The distinction between religiosity and spirituality is discussed in terms of the type-of-crime hypothesis. PMID- 16249399 TI - Ethnicity, gender, and global risk indicators in the lives of status offenders coming to the attention of the juvenile court. AB - The present study examined the risks and needs of status offenders in comparison to an at-large juvenile court sample, with specific attention paid to factors related to gender and ethnicity. Status offenders displayed significantly higher risk scores than the comparison sample in domains associated with both family and parenting concerns and educational concerns. In addition, females displayed higher risk levels than males and White youth displayed significantly higher risk levels than minority youth across a wide variety of risk domains. Further analyses were conducted regarding the participation of African American males and females in a program designed as an alternative to detention. For both groups, successful program completion was accompanied by little or no further court involvement, whereas failure to complete the program was related to a variety of negative outcomes. However, African American females and males successfully completing program requirements displayed different risks and needs at the outset of their participation. PMID- 16249400 TI - Rural and urban differences in the commission of animal cruelty. AB - Despite the recent surge in society's interest in human violence, relatively few studies have been conducted examining the closely related phenomenon of animal cruelty. Although several researchers have begun to identify some of the correlates of animal cruelty, few have attempted to understand how differences in the backgrounds of rural and urban residents have led to their abuse of animals. Using survey data from 261 inmates, the authors investigate how demographic, familial differences and species type have contributed to the frequency of acts of animal cruelty. In general, early exposure to animal abuse is a strong predictor of the subsequent behavior. However, rural inmates learned to be cruel by watching family members exclusively, whereas urban inmates learned from family members and friends. Moreover, urban inmates chose dogs, cats, and wild animals as their target animals; however, rural inmates chose only cats. PMID- 16249403 TI - Electrifying phosphatases. AB - Ci-VSP, a recently described protein with sequence similarity to both the voltage sensing domain of a voltage-gated potassium channel and the phosphatase PTEN, functions as a transmembrane phosphoinositide phosphatase that is regulated by changes in voltage across the plasma membrane. Ci-VSP dephosphorylated phosphatidylinositol-3,4,5-trisphosphate [PtdIns(3,4,5)P3] in vitro, exhibited capacitative currents that resembled ion channel gating currents, and, when coexpressed with potassium channels that are regulated by PtdIns(4,5)P2, conferred sensitivity of potassium current amplitude to prolonged changes in membrane potential. How the voltage-sensing (VS) domain of Ci-VSP communicates with the phosphatase domain, and how the VS domain moves its charges across the membrane electric field in the absence of a transmembrane pore domain, remain to be determined. PMID- 16249404 TI - The cover. James Peale (the Lamplight Portrait). PMID- 16249405 TI - Meth lab fires put heat on burn centers. PMID- 16249406 TI - Does the metabolic syndrome really exist? Diabetes and heart disease groups spar over issue. PMID- 16249407 TI - European researchers, drug companies join forces against rare diseases. PMID- 16249408 TI - Equine influenza jumps to canines. PMID- 16249409 TI - Quality improvement organizations and hospital care. PMID- 16249410 TI - Quality improvement organizations and hospital care. PMID- 16249411 TI - Quality improvement organizations and hospital care. PMID- 16249412 TI - Quality improvement organizations and hospital care. PMID- 16249413 TI - Mechanical reperfusion more than 12 hours after acute myocardial infarction. PMID- 16249414 TI - Mechanical reperfusion more than 12 hours after acute myocardial infarction. PMID- 16249415 TI - Helminth infection during pregnancy and development of infantile eczema. PMID- 16249416 TI - Antibiotic prescribing for lower respiratory tract infection. PMID- 16249417 TI - Supplemental perioperative oxygen and the risk of surgical wound infection: a randomized controlled trial. AB - CONTEXT: Supplemental perioperative oxygen has been variously reported to halve or double the risk of surgical wound infection. OBJECTIVE: To test the hypothesis that supplemental oxygen reduces infection risk in patients following colorectal surgery. DESIGN, SETTING, AND PATIENTS: A double-blind, randomized controlled trial of 300 patients aged 18 to 80 years who underwent elective colorectal surgery in 14 Spanish hospitals from March 1, 2003, to October 31, 2004. Wound infections were diagnosed by blinded investigators using Centers for Disease Control and Prevention criteria. Baseline patient characteristics, anesthetic treatment, and potential confounding factors were recorded. INTERVENTIONS: Patients were randomly assigned to either 30% or 80% fraction of inspired oxygen (FIO2) intraoperatively and for 6 hours after surgery. Anesthetic treatment and antibiotic administration were standardized. MAIN OUTCOME MEASURES: Any surgical site infection (SSI); secondary outcomes included return of bowel function and ability to tolerate solid food, ambulation, suture removal, and duration of hospitalization. RESULTS: A total of 143 patients received 30% perioperative oxygen and 148 received 80% perioperative oxygen. Surgical site infection occurred in 35 patients (24.4%) administered 30% FIO2 and in 22 patients (14.9%) administered 80% FIO2 (P=.04). The risk of SSI was 39% lower in the 80% FIO2 group (relative risk [RR], 0.61; 95% confidence interval [CI], 0.38-0.98) vs the 30% FIO2 group. After adjustment for important covariates, the RR of infection in patients administered supplemental oxygen was 0.46 (95% CI, 0.22-0.95; P = .04). None of the secondary outcomes varied significantly between the 2 treatment groups. CONCLUSIONS: Patients receiving supplemental inspired oxygen had a significant reduction in the risk of wound infection. Supplemental oxygen appears to be an effective intervention to reduce SSI in patients undergoing colon or rectal surgery. Trial Registration ClinicalTrials.gov Identifier: NCT00235456. PMID- 16249418 TI - Changing epidemiology of invasive pneumococcal disease among older adults in the era of pediatric pneumococcal conjugate vaccine. AB - CONTEXT: A conjugate vaccine targeting 7 pneumococcal serotypes was licensed for young children in 2000. In contrast to the 23-valent polysaccharide vaccine used in adults, the 7-valent conjugate vaccine affects pneumococcal carriage and transmission. Early after its introduction, incidence of invasive pneumococcal disease declined among older adults, a group at high risk for pneumococcal disease. OBJECTIVE: To determine among adults aged 50 years or older whether incidence of invasive pneumococcal disease, disease characteristics, or the spectrum of patients acquiring these illnesses have changed over the 4 years since pneumococcal conjugate vaccine licensure. DESIGN, SETTING, AND POPULATION: Population-based surveillance of invasive pneumococcal disease in 8 US geographic areas (total population, 18,813,000), 1998-2003. MAIN OUTCOME MEASURES: Incidence of invasive pneumococcal disease by pneumococcal serotype and other characteristics; frequency among case patients of comorbid conditions and other factors influencing mortality. RESULTS: Incidence of invasive pneumococcal disease among adults aged 50 years or older declined 28% (95% confidence interval [CI], -31% to -24%), from 40.8 cases/100,000 in 1998-1999 to 29.4 in 2002-2003. Among those aged 65 years or older, the 2002-2003 rate (41.7 cases/100,000) was lower than the Healthy People 2010 goal (42 cases/100,000). Among adults aged 50 years or older, incidence of disease caused by the 7 conjugate vaccine serotypes declined 55% (95% CI, -58% to -51%) from 22.4 to 10.2 cases/100,000. In contrast, disease caused by any of the 16 serotypes only in polysaccharide vaccine did not change, and disease caused by serotypes not in either vaccine increased somewhat, from 6.0 to 6.8 cases/100,000 (13%; 95% CI, 1% to 27%). Between 1998-1999 and 2002-2003, the proportion of case-patients with human immunodeficiency virus infection increased from 1.7% (47/2737) to 5.6% (124/2231) (P<.001), and those with any comorbid condition that is an indication for pneumococcal polysaccharide vaccination increased from 62.3% (1842/2955) to 72.0% (1721/2390) (P<.001). CONCLUSIONS: Our findings indicate that use of conjugate vaccine in children has substantially benefited older adults. However, persons with certain comorbid conditions may benefit less than healthier persons from the indirect effects of the new vaccine. PMID- 16249419 TI - Relevance of urine telomerase in the diagnosis of bladder cancer. AB - CONTEXT: The identification of new molecular markers is one of the most challenging goals for the early detection of bladder cancer because available noninvasive methods have neither sufficient sensitivity nor specificity to be acceptable for routine use. OBJECTIVE: To develop a relatively simple, inexpensive, and accurate test that measures telomerase activity in voided urine to apply to large-scale screening programs for bladder cancer detection. DESIGN, SETTING, AND PARTICIPANTS: Case-control study conducted in 218 men (84 healthy individuals and 134 patients at first diagnosis of histologically confirmed bladder cancer), frequency matched by age and recruited between March 2003 and November 2004 in Italy. Urine telomerase activity was determined using a highly sensitive telomeric repeat amplification protocol (TRAP) assay. Urine samples were processed for cytological diagnosis and TRAP assay. The diagnosis of bladder cancer was based on bioptic and cystoscopic examinations. The performance of the TRAP assay to detect urine telomerase activity was compared with urine cytology as an aid to early cancer detection. Quantification of urine telomerase activity was conducted in a blinded manner. MAIN OUTCOME MEASURE: Sensitivity and specificity of TRAP to detect bladder cancer. RESULTS: Using a 50 arbitrary enzymatic unit cutoff value, we validated the results obtained in the pilot study. In the overall series, sensitivity was 90% (95% confidence interval [CI], 83%-94%) and specificity was 88% (95% CI, 79%-93%). Specificity increased to 94% (95% CI, 85%-98%) for individuals aged 75 years or younger. The same predictive capacity of telomerase activity levels was observed for patients with low-grade tumors or with negative cytology results. CONCLUSIONS: The present validation study demonstrated the ability of urine telomerase activity levels to accurately detect the presence of bladder tumors in men. This test represents a potentially useful noninvasive diagnostic innovation for bladder cancer detection in high risk groups such as habitual smokers or in symptomatic patients. PMID- 16249421 TI - Suicide prevention strategies: a systematic review. AB - CONTEXT: In 2002, an estimated 877,000 lives were lost worldwide through suicide. Some developed nations have implemented national suicide prevention plans. Although these plans generally propose multiple interventions, their effectiveness is rarely evaluated. OBJECTIVES: To examine evidence for the effectiveness of specific suicide-preventive interventions and to make recommendations for future prevention programs and research. DATA SOURCES AND STUDY SELECTION: Relevant publications were identified via electronic searches of MEDLINE, the Cochrane Library, and PsychINFO databases using multiple search terms related to suicide prevention. Studies, published between 1966 and June 2005, included those that evaluated preventative interventions in major domains; education and awareness for the general public and for professionals; screening tools for at-risk individuals; treatment of psychiatric disorders; restricting access to lethal means; and responsible media reporting of suicide. DATA EXTRACTION: Data were extracted on primary outcomes of interest: suicidal behavior (completion, attempt, ideation), intermediary or secondary outcomes (treatment seeking, identification of at-risk individuals, antidepressant prescription/use rates, referrals), or both. Experts from 15 countries reviewed all studies. Included articles were those that reported on completed and attempted suicide and suicidal ideation; or, where applicable, intermediate outcomes, including help-seeking behavior, identification of at-risk individuals, entry into treatment, and antidepressant prescription rates. We included 3 major types of studies for which the research question was clearly defined: systematic reviews and meta-analyses (n = 10); quantitative studies, either randomized controlled trials (n = 18) or cohort studies (n = 24); and ecological, or population- based studies (n = 41). Heterogeneity of study populations and methodology did not permit formal meta-analysis; thus, a narrative synthesis is presented. DATA SYNTHESIS: Education of physicians and restricting access to lethal means were found to prevent suicide. Other methods including public education, screening programs, and media education need more testing. CONCLUSIONS: Physician education in depression recognition and treatment and restricting access to lethal methods reduce suicide rates. Other interventions need more evidence of efficacy. Ascertaining which components of suicide prevention programs are effective in reducing rates of suicide and suicide attempt is essential in order to optimize use of limited resources. PMID- 16249420 TI - Predictors and prevalence of paraganglioma syndrome associated with mutations of the SDHC gene. AB - CONTEXT: Paraganglioma syndrome includes inherited head and neck paragangliomas (HNPs) and adrenal or extra-adrenal pheochromocytomas and are classified according to the susceptibility genes SDHB, SDHC, and SDHD. In contrast with those with germline mutations of the SDHB and SDHD genes, clinical and genetic data on patients with mutations of SDHC are scarce. OBJECTIVE: To determine the prevalence and clinical characteristics of SDHC mutation carriers compared with patients with SDHB and SDHD mutations and with sporadic cases. DESIGN, SETTING, AND PATIENTS: Genetic screening for SDHC mutations in an international HNP registry of 121 unrelated index cases and in 371 sporadic cases from a pheochromocytoma registry, conducted January 1, 2001, until December 31, 2004. Identified index cases and affected relatives were clinically evaluated. MAIN OUTCOME MEASURES: Prevalence of and clinical findings for SDHC mutation associated HNPs vs those with SDHB and SDHD mutations. RESULTS: The prevalence of SDHC carriers was 4% in HNP but 0% in pheochromocytoma index cases. None of the SDHC mutation carriers had signs of pheochromocytoma. We compared HNPs in 22 SDHC mutation carriers with the HNPs of SDHB (n = 15) and SDHD (n = 42) mutation carriers and with 90 patients with sporadic HNPs. Location, number of tumors, malignancy, and age were different: more carotid body tumors were found in SDHC (13/22 [59%]) than in sporadic HNPs (29/90 [32%], P = .03), as well as fewer instances of multiple tumors in SDHC (2/22) than in SDHD (24/42; P<.001), 0 malignant tumors in SDHC vs 6/15 in SDHB (P = .002), and younger age at diagnosis in SDHC than in sporadic HNPs (45 vs 52 years; P = .03). CONCLUSIONS: Patients with HNP, but not those with pheochromocytoma, harbor SDHC mutations in addition to those in SDHB and SDHD. In total, more than one quarter of HNP patients carry a mutation in 1 of these 3 genes. Head and neck paragangliomas associated with SDHC mutations are virtually exclusively benign and seldom multifocal. Analysis for germline mutations of SDHC is recommended in apparently sporadic HNP to identify risk of inheritance. PMID- 16249422 TI - The patents-based pharmaceutical development process: rationale, problems, and potential reforms. AB - The pharmaceutical industry is facing substantial criticism from many directions, including financial barriers to access to drugs in both developed and developing countries, high profits, spending on advertising and marketing, and other issues. Underlying these criticisms are fundamental questions about the value of the current patent-based drug development system. Six major problems with the patent system are (1) recovery of research costs by patent monopoly reduces access to drugs; (2) market demand rather than health needs determines research priorities; (3) resources between research and marketing are misallocated; (4) the market for drugs has inherent market failures; (5) overall investment in drug research and development is too low, compared with profits; and (6) the existing system discriminates against US patients. Potential solutions fall into 3 categories: change in drug pricing through either price controls or tiered pricing; change in drug industry structure through a "buy-out" pricing system or with the public sector acting as exclusive research funder; and change in development incentives through a disease burden incentive system, orphan drug approaches, or requiring new drugs to demonstrate improvement over existing products prior to US Food and Drug Administration approval. We recommend 4 complementary reforms: (1) having no requirement to test new drug products against existing products prior to approval but requiring rigorous comparative postapproval testing; (2) international tiered pricing and systematic safeguards to prevent flow-back; (3) increased government funded research and buy-out for select conditions; and (4) targeted experiments using other approaches for health conditions in which there has been little progress and innovation over the last few decades. PMID- 16249423 TI - Improving relations between attorneys and physicians. PMID- 16249425 TI - Increasing inspired oxygen to decrease surgical site infection: time to shift the quality improvement research paradigm. PMID- 16249424 TI - Provision or distribution of growth hormone for "antiaging": clinical and legal issues. PMID- 16249426 TI - JAMA patient page. Wound infections. PMID- 16249427 TI - Diabetes and insulin secretion: the ATP-sensitive K+ channel (K ATP) connection. AB - The ATP-sensitive K+ channel (K ATP channel) senses metabolic changes in the pancreatic beta-cell, thereby coupling metabolism to electrical activity and ultimately to insulin secretion. When K ATP channels open, beta-cells hyperpolarize and insulin secretion is suppressed. The prediction that K ATP channel "overactivity" should cause a diabetic state due to undersecretion of insulin has been dramatically borne out by recent genetic studies implicating "activating" mutations in the Kir6.2 subunit of K ATP channel as causal in human diabetes. This article summarizes the emerging picture of K ATP channel as a major cause of neonatal diabetes and of a polymorphism in K ATP channel (E23K) as a type 2 diabetes risk factor. The degree of K ATP channel "overactivity" correlates with the severity of the diabetic phenotype. At one end of the spectrum, polymorphisms that result in a modest increase in K ATP channel activity represent a risk factor for development of late-onset diabetes. At the other end, severe "activating" mutations underlie syndromic neonatal diabetes, with multiple organ involvement and complete failure of glucose-dependent insulin secretion, reflecting K ATP channel "overactivity" in both pancreatic and extrapancreatic tissues. PMID- 16249428 TI - Inositol (1,4,5)-trisphosphate dynamics and intracellular calcium oscillations in pancreatic beta-cells. AB - Glucose-stimulated insulin secretion is associated with transients of intracellular calcium concentration ([Ca2+]i) in the pancreatic beta-cell. We tested the hypothesis that inositol (1,4,5)-trisphosphate [Ins(1,4,5)P3] [Ca2+]i release is incorporated in glucose-induced [Ca2+]i oscillations in mouse islets and MIN6 cells. We found that depletion of intracellular Ca2+ stores with thapsigargin increased the oscillation frequency by twofold and inhibited the slow recovery phase of [Ca2+]i oscillations. We employed a pleckstrin homology domain-containing fluorescent biosensor, phospholipase C partial differential pleckstrin homology domain-enhanced green fluorescent protein, to visualize Ins(1,4,5)P3 dynamics in insulin-secreting MIN6 cells and mouse islets in real time using a video-rate confocal system. In both types of cells, stimulation with carbamoylcholine (CCh) and depolarization with KCl results in an increase in Ins(1,4,5)P3 accumulation in the cytoplasm. When stimulated with glucose, the Ins(1,4,5)P3 concentration in the cytoplasm oscillates in parallel with oscillations of [Ca2+]i. Maximal accumulation of Ins(1,4,5)P3 in these oscillations coincides with the peak of [Ca2+]i and tracks changes in frequencies induced by the voltage-gated K+ channel blockade. We show that Ins(1,4,5)P3 release in insulin-secreting cells can be stimulated by depolarization-induced Ca2+ flux. We conclude that Ins(1,4,5)P3 concentration oscillates in parallel with [Ca2+]i in response to glucose stimulation, but it is not the driving force for [Ca2+]i oscillations. PMID- 16249429 TI - Effects of diabetes on ryanodine receptor Ca release channel (RyR2) and Ca2+ homeostasis in rat heart. AB - The defects identified in the mechanical activity of the hearts from type 1 diabetic animals include alteration of Ca2+ signaling via changes in critical processes that regulate intracellular Ca2+ concentration. These defects result partially from a dysfunction of cardiac ryanodine receptor calcium release channel (RyR2). The present study was designed to determine whether the properties of the Ca2+ sparks might provide insight into the role of RyR2 in the altered Ca2+ signaling in cardiomyocytes from diabetic animals when they were analyzed together with Ca2+ transients. Basal Ca2+ level as well as Ca2+-spark frequency of cardiomyoctes isolated from 5-week streptozotocin (STZ)-induced diabetic rats significantly increased with respect to aged-matched control rats. Ca2+ transients exhibited significantly reduced amplitude and prolonged time courses as well as depressed Ca2+ loading of sarcoplasmic reticulum in diabetic rats. Spatio-temporal properties of the Ca2+ sparks in cardiomyocytes isolated from diabetic rats were also significantly altered to being almost parallel to the changes of Ca2+ transients. In addition, RyR2 from diabetic rat hearts were hyperphosphorylated and protein levels of both RyR2 and FKBP12.6 depleted. These data show that STZ-induced diabetic rat hearts exhibit altered local Ca2+ signaling with increased basal Ca2+ level. PMID- 16249430 TI - Metallothionein-mediated antioxidant defense system and its response to exercise training are impaired in human type 2 diabetes. AB - Oxidative stress is implicated in diabetes complications, during which endogenous antioxidant defenses have important pathophysiological consequences. To date, the significance of endogenous antioxidants such as metallothioneins I and II (MT I+II) in type 2 diabetes remains unclear. To examine the MT-I+II-mediated antioxidant capacity and its response to exercise training in the skeletal muscle of patients with type 2 diabetes, biopsies and blood samples were taken from 13 matched subjects (type 2 diabetes n = 8, control subjects n = 5) both before and after 8 weeks of exercise training. Immunohistochemical analysis revealed reduced MT-I+II levels in the skeletal muscle of type 2 diabetic subjects compared with control subjects. Control subjects produced a robust increase of MT-I+II in response to training; however, in type 2 diabetes, MT-I+II levels remained essentially unchanged. Significantly lower levels of MT-I+II were also detected in the plasma of type 2 diabetic subjects compared with control subjects. These results suggest that, in control subjects, the MT-I+II defense system is active and inducible within skeletal muscle tissue and plasma. In type 2 diabetes, reduced levels of MT-I+II in muscle and plasma, as well as the deficient MT-I+II response to exercise, indicate that this antioxidant defense is impaired. This study presents a novel candidate in the pathogenesis of complications related to oxidative stress in type 2 diabetes. PMID- 16249431 TI - The human glomerular podocyte is a novel target for insulin action. AB - Microalbuminuria is significant both as the earliest stage of diabetic nephropathy and as an independent cardiovascular risk factor in nondiabetic subjects, in whom it is associated with insulin resistance. The link between disorders of cellular insulin metabolism and albuminuria has been elusive. Here, we report using novel conditionally immortalized human podocytes in vitro and human glomeruli ex vivo that the podocyte, the principal cell responsible for prevention of urinary protein loss, is insulin responsive and able to approximately double its glucose uptake within 15 min of insulin stimulation. Conditionally immortalized human glomerular endothelial cells do not respond to insulin, suggesting that insulin has a specific effect on the podocyte in the glomerular filtration barrier. The insulin response of the podocyte occurs via the facilitative glucose transporters GLUT1 and GLUT4, and this process is dependent on the filamentous actin cytoskeleton. Insulin responsiveness in this key structural component of the glomerular filtration barrier may have central relevance for understanding of diabetic nephropathy and for the association of albuminuria with states of insulin resistance. PMID- 16249433 TI - Noninvasive monitoring of diabetes-induced cutaneous nerve fiber loss and hypoalgesia in thy1-YFP transgenic mice. AB - Progressive loss of pain perception and cutaneous nerve fibers are frequently observed in diabetic patients. We evaluated the feasibility of using thy1-YFP mice that express the yellowish-green fluorescent protein (YFP) in all of their sensory/motor neurons for noninvasive monitoring of cutaneous nerve fiber loss during diabetes. Fluorescent fibers in skin sections from the leg of thy1-YFP mice stained positive for the neuron-specific protein gene product 9.5 (PGP9.5), indicating that the cutaneous fluorescent fibers are indeed nerve fibers. In diabetic thy1-YFP mice, significant small cutaneous nerve fiber loss in the leg was observed at 3 months following the onset of diabetes, but loss of heat induced pain perception occurred as early as 1 month following the onset of diabetes, indicating that functional impairment of sensory nerves precedes cutaneous nerve fiber loss. Immunostaining of skin sections of mice killed at 6 months following the onset of diabetes showed that parallel to the loss of small fluorescent nerve fibers, there was a significant decrease in fibers stained positive for calcitonin gene-related peptide, substance P, and purinoreceptor subtype in diabetic thy1-YFP mice. These mice will be useful for noninvasive monitoring of cutaneous nerve fiber degeneration and loss of heat-induced pain perception during diabetes and for the assessment of efficacy of therapeutic treatment of diabetic neuropathy. PMID- 16249434 TI - Aldose reductase deficiency prevents diabetes-induced blood-retinal barrier breakdown, apoptosis, and glial reactivation in the retina of db/db mice. AB - In 15-month-old db/db mice, signs of diabetic retinopathy, including blood retinal barrier breakdown, loss of pericytes, neuro-retinal apoptosis, glial reactivation, and proliferation of blood vessels, were evident. These changes in the diabetic retina were associated with increased expression of aldose reductase (AR). To further understand the role of AR in the pathogenesis of diabetic retinopathy, we generated db/db mice with an AR null mutation (AR-/- db/db). AR deficiency led to fewer retinal blood vessels with IgG leakage, suggesting that AR may contribute to blood-retinal barrier breakdown. AR deficiency also prevented diabetes-induced reduction of platelet/endothelial cell adhesion molecule-1 expression and increased expression of vascular endothelial growth factor, which may have contributed to blood-retinal barrier breakdown. In addition, long-term diabetes-induced neuro-retinal stress and apoptosis and proliferation of blood vessels were less prominent in AR-/- db/db mice. These findings indicate that AR is responsible for the early events in the pathogenesis of diabetic retinopathy, leading to a cascade of retinal lesions, including blood retinal barrier breakdown, loss of pericytes, neuro-retinal apoptosis, glial reactivation, and neovascularization. PMID- 16249432 TI - Glycation and carboxymethyllysine levels in skin collagen predict the risk of future 10-year progression of diabetic retinopathy and nephropathy in the diabetes control and complications trial and epidemiology of diabetes interventions and complications participants with type 1 diabetes. AB - Several mechanistic pathways linking hyperglycemia to diabetes complications, including glycation of proteins and formation of advanced glycation end products (AGEs), have been proposed. We investigated the hypothesis that skin collagen glycation and AGEs predict the risk of progression of microvascular disease. We measured glycation products in the skin collagen of 211 Diabetes Control and Complications Trial (DCCT) volunteers in 1992 who continued to be followed in the Epidemiology of Diabetes Interventions and Complications study for 10 years. We determined whether the earlier measurements of glycated collagen and AGE levels correlated with the risk of progression of retinopathy and nephropathy from the end of the DCCT to 10 years later. In multivariate analyses, the combination of furosine (glycated collagen) and carboxymethyllysine (CML) predicted the progression of retinopathy (chi2 = 59.4, P < 0.0001) and nephropathy (chi2 = 18.2, P = 0.0001), even after adjustment for mean HbA(1c) (A1C) (chi2 = 32.7, P < 0.0001 for retinopathy) and (chi2 = 12.8, P = 0.0016 for nephropathy). The predictive effect of A1C vanished after adjustment for furosine and CML (chi2 = 0.0002, P = 0.987 for retinopathy and chi2 = 0.0002, P = 0.964 for nephropathy). Furosine explained more of the variation in the 10-year progression of retinopathy and nephropathy than did CML. These results strengthen the role of glycation of proteins and AGE formation in the pathogenesis of retinopathy and nephropathy. Glycation and subsequent AGE formation may explain the risk of these complications associated with prior A1C and provide a rational basis for the phenomenon of "metabolic memory" in the pathogenesis of these diabetes complications. PMID- 16249435 TI - Large genomic rearrangements in the hepatocyte nuclear factor-1beta (TCF2) gene are the most frequent cause of maturity-onset diabetes of the young type 5. AB - Maturity-onset diabetes of the young (MODY) 5 is caused by mutations in the TCF2 gene encoding the transcription factor hepatocyte nuclear factor-1beta. However, in 60% of the patients with a phenotype suggesting MODY5, no point mutation is detected in TCF2. We have hypothesized that large genomic rearrangements of TCF2 that are missed by conventional screening methods may account for this observation. In 40 unrelated patients presenting with MODY5 phenotype, TCF2 was screened for mutations by sequencing. Patients without mutations were then screened for TCF2 rearrangements by the quantitative multiplex PCR of short fluorescent fragments (QMPSF). Among the 40 patients, the overall detection rate was 70%: 18 had point mutations, 9 had whole-gene deletions, and 1 had a deletion of a single exon. Similar phenotypes were observed in patients with mutations and in subjects with large deletions. These results suggest that MODY5 is more prevalent than previously reported, with one-third of the cases resulting from large deletions of TCF2. Because QMPSF is more rapid and cost effective than sequencing, we propose that patients whose phenotype is consistent with MODY5 should be screened first with the QMPSF assay. In addition, other MODY genes should be screened for large genomic rearrangements. PMID- 16249436 TI - Partial gene deletion of heart-type fatty acid-binding protein limits the severity of dietary-induced insulin resistance. AB - The aim of this study was to determine the contribution of heart-type fatty acid binding protein (H-FABP) to glucose and long-chain fatty acid (LCFA) utilization in dietary-induced insulin resistance. We tested the hypothesis that H-FABP facilitates increases in LCFA flux present in glucose-intolerant states and that a partial reduction in the amount of this protein would compensate for all or part of the impairment. Transgenic H-FABP heterozygotes (HET) and wild-type (WT) littermates were studied following chow diet (CHD) or high-fat diet (HFD) for 12 weeks. Catheters were surgically implanted in the carotid artery and jugular vein for sampling and infusions, respectively. Following 5 days of recovery, mice received either a saline infusion or underwent a euglycemic insulin clamp (4 mU x kg(-1) x min(-1)) for 120 min. At 90 min, a bolus of 2-deoxyglucose and [125I]-15 (rho-iodophenyl)-3-R,S-methylpentadecanoic acid were administered to obtain indexes of glucose and LCFA utilization. At 120 min, skeletal muscles were excised for tracer determination. All HFD mice were obese and hyperinsulinemic; however, only HFD-WT mice were hyperglycemic. Glucose infusion rates during insulin clamps were 49 +/- 4, 59 +/- 4, 16 +/- 4, and 33 +/- 4 mg x kg(-1) x min( 1) for CHD-WT, CHD-HET, HFD-WT, and HFD-HET mice, respectively, showing that HET limited the severity of whole-body insulin resistance with HFD. Insulin stimulated muscle glucose utilization was attenuated in HFD-WT but unaffected in HFD-HET mice. Conversely, rates of LCFA clearance were increased with HFD feeding in HFD-WT but not in HFD-HET mice. In conclusion, a partial reduction in H-FABP protein normalizes fasting glucose levels and improves whole-body insulin sensitivity in HFD-fed mice despite obesity. PMID- 16249437 TI - Liver markers and development of the metabolic syndrome: the insulin resistance atherosclerosis study. AB - Nonalcoholic fatty liver disease (NAFLD) is emerging as a component of the metabolic syndrome, although it is not known whether markers of NAFLD, including elevated concentrations of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALK), predict the development of metabolic syndrome. Our objective was to investigate the associations of elevated AST, ALT, and other liver markers, including C-reactive protein (CRP), with incident National Cholesterol Education Program-defined metabolic syndrome among 633 subjects in the Insulin Resistance Atherosclerosis Study who were free of metabolic syndrome at baseline. Insulin sensitivity (Si) and acute insulin response (AIR) were directly measured from the frequently sampled intravenous glucose tolerance test among African-American, Hispanic, and non-Hispanic white subjects aged 40-69 years. After 5.2 years, 127 individuals had developed metabolic syndrome. In separate logistic regression models adjusting for age, sex, ethnicity, clinic, and alcohol consumption, subjects in the upper quartiles of ALT, ALK, and CRP were at significantly increased risk of incident metabolic syndrome compared with those in the lowest quartile: ALT, odds ratio 2.50 (95% CI 1.38-4.51); ALK, 2.28 (1.24-4.20); and CRP, 1.33 (1.09-1.63). Subjects in the upper quartile of the AST-to-ALT ratio were at significantly reduced metabolic syndrome risk (0.40 [0.22-0.74]). After further adjustment for waist circumference, Si, AIR, and impaired glucose tolerance, the associations of ALT and the AST-to-ALT ratio with incident metabolic syndrome remained significant (ALT, 2.12 [1.10-4.09]; the AST-to-ALT ratio, 0.48 [0.25-0.95]). These associations were not modified by ethnicity or sex, and they remained significant after exclusion of former and heavy drinkers. In conclusion, NAFLD markers ALT and the AST-to-ALT ratio predict metabolic syndrome independently of potential confounding variables, including directly measured Si and AIR. PMID- 16249438 TI - Effect of a sustained reduction in plasma free fatty acid concentration on intramuscular long-chain fatty Acyl-CoAs and insulin action in type 2 diabetic patients. AB - To investigate the effect of a sustained (7-day) decrease in plasma free fatty acid (FFA) concentrations on insulin action and intramyocellular long-chain fatty acyl-CoAs (LCFA-CoAs), we studied the effect of acipimox, a potent inhibitor of lipolysis, in seven type 2 diabetic patients (age 53 +/- 3 years, BMI 30.2 +/- 2.0 kg/m2, fasting plasma glucose 8.5 +/- 0.8 mmol/l, HbA 1c 7.5 +/- 0.4%). Subjects received an oral glucose tolerance test (OGTT) and 120-min euglycemic insulin (80 mU/m2 per min) clamp with 3-[3H]glucose/vastus lateralis muscle biopsies to quantitate rates of insulin-mediated whole-body glucose disposal (Rd) and intramyocellular LCFA-CoAs before and after acipimox (250 mg every 6 h for 7 days). Acipimox significantly reduced fasting plasma FFAs (from 563 +/- 74 to 230 +/- 33 micromol/l; P < 0.01) and mean plasma FFAs during the OGTT (from 409 +/- 44 to 184 +/- 22 micromol/l; P < 0.01). After acipimox, decreases were seen in fasting plasma insulin (from 78 +/- 18 to 42 +/- 6 pmol/l; P < 0.05), fasting plasma glucose (from 8.5 +/- 0.8 to 7.0 +/- 0.5 mmol/l; P < 0.02), and mean plasma glucose during the OGTT (from 14.5 +/- 0.8 to 13.0 +/- 0.8 mmol/l; P < 0.05). After acipimox, insulin-stimulated Rd increased from 3.3 +/- 0.4 to 4.4 +/ 0.4 mg x kg(-1) x min(-1) (P < 0.03), whereas suppression of endogenous glucose production (EGP) was similar and virtually complete during both insulin clamp studies (0.16 +/- 0.10 vs. 0.14 +/- 0.10 mg x kg(-1) x min(-1); P > 0.05). Basal EGP did not change after acipimox (1.9 +/- 0.2 vs. 1.9 +/- 0.2 mg x kg(-1) x min( 1)). Total muscle LCFA-CoA content decreased after acipimox treatment (from 7.26 +/- 0.58 to 5.64 +/- 0.79 nmol/g; P < 0.05). Decreases were also seen in muscle palmityl CoA (16:0; from 1.06 +/- 0.10 to 0.75 +/- 0.11 nmol/g; P < 0.05), palmitoleate CoA (16:1; from 0.48 +/- 0.05 to 0.33 +/- 0.05 nmol/g; P = 0.07), oleate CoA (18:1; from 2.60 +/- 0.11 to 1.95 +/- 0.31 nmol/g; P < 0.05), linoleate CoA (18:2; from 1.81 +/- 0.26 to 1.38 +/- 0.18 nmol/g; P = 0.13), and linolenate CoA (18:3; from 0.27 +/- 0.03 to 0.19 +/- 0.02 nmol/g; P < 0.03) levels after acipimox treatment. Muscle stearate CoA (18:0) did not decrease after acipimox treatment. The increase in R(d) correlated strongly with the decrease in muscle palmityl CoA (r = 0.75, P < 0.05), oleate CoA (r = 0.76, P < 0.05), and total muscle LCFA-CoA (r = 0.74, P < 0.05) levels. Plasma adiponectin did not change significantly after acipimox treatment (7.9 +/- 1.8 vs. 7.5 +/- 1.5 microg/ml). These data demonstrate that the reduction in intramuscular LCFA CoA content is closely associated with enhanced insulin sensitivity in muscle after a chronic reduction in plasma FFA concentrations in type 2 diabetic patients despite the lack of an effect on plasma adiponectin concentration. PMID- 16249439 TI - The stimulatory effect of globular adiponectin on insulin-stimulated glucose uptake and fatty acid oxidation is impaired in skeletal muscle from obese subjects. AB - Adiponectin is an adipose-derived hormone that plays an important role in regulating insulin sensitivity in rodents. However, little is known regarding the effect of adiponectin on metabolism in human skeletal muscle. Therefore, we examined whether the globular head of adiponectin, gAcrp30, acutely activates fatty acid oxidation and glucose uptake in isolated human skeletal muscle. Furthermore, we aimed to determine whether these effects would differ in muscle from lean versus obese individuals. Treatment with gAcrp30 (2.5 microg/ml) increased fatty acid oxidation in lean muscle (70%, P < 0.0001) and to a lesser extent in obese muscle (30%, P < 0.01). In the absence of insulin, gAcrp30 increased glucose uptake 37% in lean (P < 0.05) and 33% in obese muscle (P < 0.05). Combined exposure of insulin and gAcrp30 demonstrated an additive effect on glucose uptake in lean and obese individuals, but this effect was reduced by 50% in obese muscle (P < 0.05). These metabolic effects were attributable to an increase in AMP-activated protein kinase-alpha1 (AMPKalpha1) and AMPKalpha2 activity. However, in obese muscle the activation of AMPKalpha2 by gAcrp30 was blunted. This study provides evidence that gAcrp30 plays a role in regulating fatty acid and glucose metabolism in human skeletal muscle. However, the effects are blunted in obesity, indicating the possible development of adiponectin resistance. PMID- 16249440 TI - Insulin-stimulated glucose uptake does not require p38 mitogen-activated protein kinase in adipose tissue or skeletal muscle. AB - It has been proposed that p38 mitogen-activated protein kinase (MAPK) isoforms sensitive to the pyridinylimidazole compounds SB 203580 and SB 202190 may participate in the acute insulin-dependent activation of glucose transporters recruited to the plasma membrane of adipocytes and skeletal muscle. Here, we explore whether these kinases support the insulin stimulation of glucose uptake in these tissues by investigating the effects of a genetic loss in p38beta and that of the p38 MAPK inhibitor SB 203580. Glucose uptake in adipocytes and soleus muscle was stimulated by insulin by up to fourfold irrespective of whether tissues were isolated from wild-type or p38beta-null mice. Consistent with this finding, mice lacking p38beta exhibited normal glucose tolerance, insulinemia, and glycemia compared with their wild-type counterparts. Insulin-stimulated glucose uptake was not inhibited by SB 203580 when adipocytes were preincubated with the drug at a cytocrit of 50%, but intriguingly, uptake was suppressed (by 35%) when the cytocrit was reduced by one-half. Despite the activation of glucose uptake at the higher cytocrit, insulin failed to induce any detectable activation of p38 MAPK, whereas p38 signaling was robustly activated by anisomycin in a SB 203580-sensitive manner. Although insulin also failed to induce any detectable activation of p38 MAPK in muscle, insulin-dependent glucose uptake was reduced by SB 203580 (approximately 44%) in muscle of both wild-type and p38beta-null mice. Our results indicate that p38beta is not required for insulin-stimulated glucose uptake in adipocytes or muscle. Moreover, given that insulin fails to promote any significant activation of p38 MAPK in these tissues and the finding that sensitivity of glucose uptake, but not that of the kinase, to SB 203580 can be influenced by cytocrit, we suggest that p38 signaling is unlikely to participate in any putative activation of transporters recruited to the cell surface by insulin and that SB 203580 suppresses insulin-stimulated glucose transport by a mechanism unrelated to its inhibitory effect on p38 MAPK. PMID- 16249441 TI - Activation of ATP-sensitive K+ channels in the ventromedial hypothalamus amplifies counterregulatory hormone responses to hypoglycemia in normal and recurrently hypoglycemic rats. AB - The mechanism(s) by which glucosensing neurons detect fluctuations in glucose remains largely unknown. In the pancreatic beta-cell, ATP-sensitive K+ channels (K ATP channels) play a key role in glucosensing by providing a link between neuronal metabolism and membrane potential. The present study was designed to determine in vivo whether the pharmacological opening of ventromedial hypothalamic K ATP channels during systemic hypoglycemia would amplify hormonal counterregulatory responses in normal rats and those with defective counterregulation arising from prior recurrent hypoglycemia. Controlled hypoglycemia (approximately 2.8 mmol/l) was induced in vivo using a hyperinsulinemic (20 mU x kg(-1) x min(-1)) glucose clamp technique in unrestrained, overnight-fasted, chronically catheterized Sprague-Dawley rats. Immediately before the induction of hypoglycemia, the rats received bilateral ventromedial hypothalamic microinjections of either the potassium channel openers (KCOs) diazoxide and NN414 or their respective controls. In normal rats, both KCOs amplified epinephrine and glucagon counterregulatory responses to hypoglycemia. Moreover, diazoxide also amplified the counterregulatory responses in a rat model of defective hormonal counterregulation. Taken together, our data suggest that the K ATP channel plays a key role in vivo within glucosensing neurons in the ventromedial hypothalamus in the detection of incipient hypoglycemia and the initiation of protective counterregulatory responses. We also conclude that KCOs may offer a future potential therapeutic option for individuals with insulin-treated diabetes who develop defective counterregulation. PMID- 16249442 TI - Impaired generation of reactive oxygen species in leprechaunism through downregulation of Nox4. AB - Leprechaunism features a clinical constellation characterized by extreme insulin resistance, growth retardation, and several distinct developmental abnormalities. One puzzling observation about leprechaunism is that mutations in the insulin receptor gene frequently associated with this syndrome cannot account for the aberrant responses of cultured cells to other growth factors. Here we report that the generation of reactive oxygen species (ROS) is impaired in cells from leprechaunism patients, thus shedding new light on this issue. Stimulation of patients' skin fibroblast cells with platelet-derived growth factor (PDGF) resulted in a lower-level tyrosine phosphorylation of cytosolic proteins compared with that seen in normal cells. In addition, consistent with the hypothesis that ROS mediate the level of tyrosine phosphorylation of cytosolic proteins through inactivation of protein tyrosine phosphatases (PTPases), patient fibroblast cells showed a significantly higher phosphatase activity than normal cells. We further showed that the lower-level tyrosine phosphorylation in response to growth factors results from the downregulation of an NADPH oxidase, Nox4, which in turn results in the reduction of ROS generation. Ectopic expression of Nox4 in the patient fibroblast cells consistently restored PDGF-induced ROS production and regulation of PTPase activities. Taken together, these data provide insight into the mechanisms through which growth retardation is associated with leprechaunism syndrome. PMID- 16249443 TI - Central leptin acutely reverses diet-induced hepatic insulin resistance. AB - Voluntary overfeeding rapidly induces resistance to the effects of systemic insulin and leptin on liver glucose metabolism. To examine whether central administration of recombinant leptin can restore leptin and insulin action on liver glucose fluxes, we infused leptin in the third cerebral ventricle of conscious overfed rats during pancreatic-insulin clamp studies. The effect of leptin on the phosphorylation of the signal transducer and activator of transcription-3 in the arcuate nuclei of the hypothalamus was similar in animals fed a regular diet or a high-fat diet for 3 days. The infusion of leptin in the third cerebral ventricle markedly inhibited glucose production in rats fed a high fat diet mainly by decreasing glycogenolysis. The inhibition of glycogenolysis was sufficient to normalize glucose production and was accompanied by leptin induced decreases in the hepatic expression of glucose-6-phosphatase and phosphoenolpyruvate carboxykinase. Thus central administration of leptin rescues the hepatic insulin resistance induced by short-term hyperphagia. PMID- 16249444 TI - Adipocyte lipases and defect of lipolysis in human obesity. AB - The mobilization of fat stored in adipose tissue is mediated by hormone-sensitive lipase (HSL) and the recently characterized adipose triglyceride lipase (ATGL), yet their relative importance in lipolysis is unknown. We show that a novel potent inhibitor of HSL does not inhibit other lipases. The compound counteracted catecholamine-stimulated lipolysis in mouse adipocytes and had no effect on residual triglyceride hydrolysis and lipolysis in HSL-null mice. In human adipocytes, catecholamine- and natriuretic peptide-induced lipolysis were completely blunted by the HSL inhibitor. When fat cells were not stimulated, glycerol but not fatty acid release was inhibited. HSL and ATGL mRNA levels increased concomitantly during adipocyte differentiation. Abundance of the two transcripts in human adipose tissue was highly correlated in habitual dietary conditions and during a hypocaloric diet, suggesting common regulatory mechanisms for the two genes. Comparison of obese and nonobese subjects showed that obesity was associated with a decrease in catecholamine-induced lipolysis and HSL expression in mature fat cells and in differentiated preadipocytes. In conclusion, HSL is the major lipase for catecholamine- and natriuretic peptide stimulated lipolysis, whereas ATGL mediates the hydrolysis of triglycerides during basal lipolysis. Decreased catecholamine-induced lipolysis and low HSL expression constitute a possibly primary defect in obesity. PMID- 16249445 TI - Peptide YY(3-36) inhibits morning, but not evening, food intake and decreases body weight in rhesus macaques. AB - Peptide YY(3-36) [PYY(3-36)] is a hormone that is released after meal ingestion that is currently being investigated for the treatment of obesity; however, there are conflicting reports of the effects of PYY(3-36) on energy balance in rodent models. To shed light on this controversy, we studied the effect of PYY(3-36) on food intake and body weight in a nonhuman primate. Intravenous PYY(3-36) infusions before a morning meal transiently suppressed the rate of food intake but did not suppress the evening meal or 24-h intake. Twice-daily or continuous intravenous PYY(3-36) infusions to supraphysiological levels (levels that exceeded normal physiological levels) again suppressed the rate of feeding for the morning but not the evening meal. Twice-daily intravenous PYY(3-36) infusions for 2 weeks significantly decreased body weight in all test animals (average weight loss 1.9%) without changing insulin response to glucose infusion. These results show that endogenous PYY(3-36) may alter morning but not evening meal intake, and supraphysiological doses are required for effective suppression of food intake. PMID- 16249447 TI - Exposure to farming environments in early life and type 1 diabetes: a case control study. AB - It has been hypothesized that a stimulation of regulatory cytokines by microbial compounds reduces autoimmune as well as atopic diseases. Farm-related contact to microbial compounds protects from allergies, but no data on the association between farm contact and type 1 diabetes is available. The aim of this study was to test this association. A case-control study was conducted in five children's hospitals. Regular contact to farm animals and potential confounders were assessed using a postal questionnaire. Eligible subjects were all prevalent cases with type 1 diabetes registered in the hospitals and hospital-based control subjects (response rate 91%). Included were children aged 6-16 years living in rural areas with German nationality (242 case and 224 control subjects). Regular contact to stables (adjusted odds ratio 1.2 [95% CI 0.5-2.7]) was not associated with type 1 diabetes. In addition, regular contact to specific farm animals was not associated with case status. There was a tendency for an inverse relationship between allergic rhinitis and type 1 diabetes (0.6 [0.3-1.1]; P = 0.11). There was no evidence that early exposure to farm animals largely decreases the risk in children for developing type 1 diabetes. PMID- 16249446 TI - Islet cell hormonal responses to hypoglycemia after human islet transplantation for type 1 diabetes. AB - Islet transplantation can eliminate severe hypoglycemic episodes in patients with type 1 diabetes; however, whether intrahepatic islets respond appropriately to hypoglycemia after transplantation has not been fully studied. We evaluated six islet transplant recipients, six type 1 diabetic subjects, and seven nondiabetic control subjects using a stepped hyperinsulinemic-hypoglycemic clamp. Also, three islet transplant recipients and the seven control subjects underwent a paired hyperinsulinemic-euglycemic clamp. In response to hypoglycemia, C-peptide was similarly suppressed in islet transplant recipients and control subjects and was not detectable in type 1 diabetic subjects. Glucagon was significantly more suppressed in type 1 diabetic subjects than in islet transplant recipients (P < 0.01), although the glucagon in islet transplant recipients failed to activate as in the control subjects (P < 0.01). Pancreatic polypeptide failed to activate in both type 1 diabetic subjects and islet transplant recipients compared with control subjects (P < 0.01). In islet transplant recipients, glucagon was suppressed normally by hyperinsulinemia during the euglycemic clamp and was significantly greater during the paired hypoglycemic clamp (P < 0.01). These results suggest that after islet transplantation and in response to insulin induced hypoglycemia, endogenous insulin secretion is appropriately suppressed and glucagon secretion may be partially restored. PMID- 16249448 TI - Mechanisms of glucose-induced secretion of pancreatic-derived factor (PANDER or FAM3B) in pancreatic beta-cells. AB - Pancreatic-derived factor (PANDER) is an islet-specific cytokine present in both pancreatic alpha- and beta-cells, which, in vitro, induces beta-cell apoptosis of primary islet and cell lines. In this study, we investigated whether PANDER is secreted by pancreatic alpha- and beta-cells and whether PANDER secretion is regulated by glucose and other insulin secretagogues. In mouse-derived insulin secreting beta-TC3 cells, PANDER secretion in the presence of stimulatory concentrations of glucose was 2.8 +/- 0.4-fold higher (P < 0.05) than without glucose. Insulin secretion was similarly increased by glucose in the same cells. The total concentration of secreted PANDER in the medium was approximately 6-10 ng/ml (0.3-0.5 nmol/l) after a 24-h culture with glucose. L-Glucose failed to stimulate PANDER secretion in beta-TC3 cells. KCl stimulated PANDER secretion 2.1 +/- 0.1-fold compared with control without glucose. An L-type Ca2+ channel inhibitor, nifedipine, completely blocked both glucose- or KCl-induced insulin and PANDER secretion. In rat-derived INS-1 cells, glucose (20 mmol/l) stimulated PANDER secretion 4.4 +/- 0.9-fold, while leucine plus glutamine stimulated 4.4 +/ 0.7-fold compared with control without glucose. In mouse islets overexpressing PANDER, glucose (20 mmol/l) stimulated PANDER secretion 3.2 +/- 0.5-fold (P < 0.05) compared with basal (3 mmol/l glucose). PANDER was also secreted by alpha TC3 cells but was not stimulated by glucose. Mutations of cysteine 229 or of cysteines 91 and 229 to serine, which may form one disulfide bond, and truncation of the COOH-terminus or NH2-terminus of PANDER all resulted in failure of PANDER secretion, even though these mutant or truncated PANDERs were highly expressed within the cells. In conclusion, we found that 1) PANDER is secreted from both pancreatic alpha- and beta-cells, 2) glucose stimulates PANDER secretion dose dependently in beta-cell lines and primary islets but not in alpha-cells, 3) PANDER is likely cosecreted with insulin via the same regulatory mechanisms, and 4) structure and conformation is vital for PANDER secretion. PMID- 16249449 TI - Restitution of defective glucose-stimulated insulin secretion in diabetic GK rat by acetylcholine uncovers paradoxical stimulatory effect of beta-cell muscarinic receptor activation on cAMP production. AB - Because acetylcholine (ACh) is a recognized potentiator of glucose-stimulated insulin release in the normal beta-cell, we have studied ACh's effect on islets of the Goto-Kakizaki (GK) rat, a spontaneous model of type 2 diabetes. We first verified that ACh was able to restore the insulin secretory glucose competence of the GK beta-cell. Then, we demonstrated that in GK islets 1) ACh elicited a first phase insulin release at low glucose, whereas it had no effect in Wistar; 2) total phospholipase C activity, ACh-induced inositol phosphate production, and intracellular free calcium concentration ([Ca2+]i) elevation were normal; 3) ACh triggered insulin release, even in the presence of thapsigargin, which induced a reduction of the ACh-induced [Ca2+]i response (suggesting that ACh produces amplification signals that augment the efficacy of elevated [Ca2+]i on GK exocytosis); 4) inhibition of protein kinase C did not affect [Ca2+]i nor the insulin release responses to ACh; and 5) inhibition of cAMP-dependent protein kinases (PKAs), adenylyl cyclases, or cAMP generation, while not affecting the [Ca2+]i response, significantly lowered the insulinotropic response to ACh (at low and high glucose). In conclusion, ACh acts mainly through activation of the cAMP/PKA pathway to potently enhance Ca2+-stimulated insulin release in the GK beta-cell and, in doing so, normalizes its defective glucose responsiveness. PMID- 16249450 TI - Is there a role for locally produced interleukin-1 in the deleterious effects of high glucose or the type 2 diabetes milieu to human pancreatic islets? AB - Different degrees of beta-cell failure and apoptosis are present in type 1 and type 2 diabetes. It has been recently suggested that high glucose-induced beta cell apoptosis in type 2 diabetes shares a final common pathway with type 1 diabetes, involving interleukin-1beta (IL-1beta) production by beta-cells, nuclear factor-kappaB (NF-kappaB) activation, and death via Fas-FasL. The aim of this study was to test whether human islet exposure to high glucose in vitro, or to the type 2 diabetes environment in vivo, induces IL-1beta expression and consequent activation of NF-kappaB-dependent genes. Human islets were isolated from five normoglycemic organ donors. The islets were cultured for 48 h to 7 days at 5.6, 11, or 28 mmol/l glucose. For comparative purposes, islets were also exposed to IL-1beta. Gene mRNA expression levels were assessed by real-time RT PCR in a blinded fashion. Culture of the human islets at 11 and 28 mmol/l glucose induced a four- to fivefold increase in medium insulin as compared with 5.6 mmol/l glucose, but neither IL-1beta nor IL-1 receptor antagonist (IL-1ra) expression changed. IL-1beta and IL-1ra protein release to the medium was also unchanged. Stimulated human monocytes, studied in parallel, released >50-fold more IL-1beta than the islets. There was also no glucose-induced islet Fas expression. Expression of the NF-kappaB-dependent genes IkappaB-alpha and monocyte chemoattractant protein (MCP)-1 was induced in human islets by IL-1beta but not by high glucose. In a second set of experiments, human islets were isolated from seven type 2 diabetic patients and eight control subjects. The findings on mRNA levels were essentially the same as in the in vitro experiments, namely the in vivo diabetic state did not induce IL-1beta, Fas, or MCP-1 expression in human islets, and also did not modify IL-1ra expression. The present findings suggest that high glucose in vitro, or the diabetic milieu in vivo, does not induce IL-1beta production or NF-kappaB activation in human islets. This makes it unlikely that locally produced IL-1beta is an important mediator of glucotoxicity to human islets and argues against the IL-1beta-NF kappaB-Fas pathway as a common mediator for beta-cell death in type 1 and type 2 diabetes. PMID- 16249452 TI - Insulin resistance, the metabolic syndrome, and incident cardiovascular events in the Framingham Offspring Study. AB - The metabolic syndrome and insulin resistance have been related to incident cardiovascular disease (CVD), but it is uncertain if metabolic syndrome predicts CVD independent of insulin resistance. Our study sample included 2,898 people without diabetes or CVD at baseline. Metabolic syndrome was defined by the National Cholesterol Education Program (NCEP) Expert Panel on Detection, Evaluation, and Treatment of High Blood Cholesterol in Adults (Adult Treatment Panel III) criteria. Insulin resistance was defined by the homeostasis model assessment (HOMA-IR) and by Gutt et al.'s insulin sensitivity index (ISI(0,120)). Age- and sex-adjusted proportional hazards regression models assessed the association of baseline metabolic syndrome and insulin resistance to 7-year CVD risk (186 events). Metabolic syndrome and both measures of insulin resistance were individually related to incident CVD (age- and sex-adjusted hazard ratio [HR] for metabolic syndrome [present versus absent]: 2.0 [95% CI 1.5-2.6], P = 0.0001; for HOMA-IR: 1.9 [1.2-2.9], P = 0.003; and for ISI(0,120) [both highest versus lowest quartile]: 0.5 [0.3-0.7], P = 0.001). In models adjusted for age, sex, LDL cholesterol, and smoking status and including metabolic syndrome, ISI(0,120) levels were independently related to incident CVD (0.5 [0.3-0.8], P = 0.004), whereas HOMA-IR levels were not (1.3 [0.8-2.1], P = 0.24); metabolic syndrome was associated with increased CVD risk in both models (HR 1.6, P < or = 0.007 in both). In conclusion, metabolic syndrome and ISI(0,120) but not HOMA-IR independently predicted incident CVD. Metabolic syndrome may not capture all the CVD risk associated with insulin resistance. PMID- 16249451 TI - Homocysteine metabolism in ZDF (type 2) diabetic rats. AB - Mild hyperhomocysteinemia is a risk factor for many diseases, including cardiovascular disease. We determined the effects of insulin resistance and of type 2 diabetes on homocysteine (Hcy) metabolism using Zucker diabetic fatty rats (ZDF/Gmi fa/fa and ZDF/Gmi fa/?). Plasma total Hcy was reduced in ZDF fa/fa rats by 24% in the pre-diabetic insulin-resistant stage, while in the frank diabetic stage there was a 59% reduction. Hepatic activities of several enzymes that play a role in the removal of Hcy:cystathionine beta-synthase (CBS), cystathionine gamma-lyase, and betaine:Hcy methyltransferase (BHMT) were increased as was methionine adenosyltransferase. CBS and BHMT mRNA levels and the hepatic level of S-adenosylmethionine were also increased in the ZDF fa/fa rats. Studies with primary hepatocytes showed that Hcy export and the transsulfuration flux in cells from ZDF fa/fa rats were particularly sensitive to betaine. Interestingly, liver betaine concentration was found to be significantly lower in the ZDf fa/fa rats at both 5 and 11 weeks. These results emphasize the importance of betaine metabolism in determining plasma Hcy levels in type 2 diabetes. PMID- 16249453 TI - Celiac-superior mesenteric ganglionectomy, but not vagotomy, suppresses the sympathoadrenal response to insulin-induced hypoglycemia. AB - Afferent innervation of the portal vein has been shown to be critical in hypoglycemic detection, but the neural pathway by which these afferents ascend remains unknown. To ascertain the role of vagal afferents versus spinal afferents in hypoglycemic detection, the catecholamine response to hypoglycemia was assessed in male Wistar rats undergoing hepatic vagotomy (HV), total subdiaphragmatic vagotomy (TSV), or celiac-superior mesenteric ganglionectomy (CSMG). After recovering from the surgery, the animals were exposed to a hyperinsulinemic-hypoglycemic clamp, with glucose infused peripherally via the jugular vein. In all animals, systemic hypoglycemia (2.64 +/- 0.03 mmol/l) was induced via jugular vein insulin infusion (25 mU x kg(-1) x min(-1)). No significant differences were observed among the groups with respect to arterial glucose or insulin concentration. When hypoglycemia was induced in sham-operated control animals, epinephrine was observed to rise from a basal value of 0.84 +/- 0.10 to 25.18 +/- 1.24 nmol/l. Neither HV nor TSV had any significant impact on the epinephrine response to hypoglycemia. In contrast, CSMG animals demonstrated a significant suppression in the epinephrine response to whole-body hypoglycemia (11.25 +/- 1.21 vs. 22.32 +/- 0.86 nmol/l in CSMG vs. controls; P < 0.05). The norepinephrine response for controls, 2.00 +/- 0.22 at basal and rising to 8.95 +/- 0.20 nmol/l in hypoglycemia, was not significantly different from that of the HV and TSV animals. As with epinephrine, the norepinephrine response to hypoglycemia was significantly suppressed in CSMG compared with control animals (4.72 +/- 0.48 vs. 7.15 +/- 0.76 nmol/l; P < 0.05). These findings are consistent with the idea that hypoglycemic detection at the portal vein is mediated by spinal, and not vagal, glucose-sensitive afferents. PMID- 16249454 TI - Two-hour seven-sample oral glucose tolerance test and meal protocol: minimal model assessment of beta-cell responsivity and insulin sensitivity in nondiabetic individuals. AB - Highly informative yet simple protocols to assess insulin secretion and action would considerably enhance the quality of epidemiological and large-scale clinical trials. In an effort to develop such protocols, a 5-h, 11-sample oral glucose tolerance test (OGTT) was performed in 100 individuals and a 7-h, 21 sample meal in another 100. Plasma glucose, insulin, and C-peptide concentrations were measured. We show that virtually the same minimal model assessment of beta cell responsivity (dynamic [Phi(d)] and static [Phi(s)]), insulin sensitivity (Si), and disposition index (DI) can be obtained with a reduced seven-sample 2-h protocol: Phi(d), reduced versus full: 871.50 vs. 873.32, r = 0.98 in OGTT and 494.88 vs. 477.99 10(-9), r = 0.91 in meal; Phi(s): 42.36 vs. 44.35, r = 0.88 in OGTT and 35.31 vs. 35.37 10(-9) min(-1), r = 0.90 in meal; Si: 24.33 vs. 22.77 10(-5) dl x kg(-1) x min(-1) per pmol/l, r = 0.89 in OGTT and 19.03 vs. 19.77 10( 5) dl x kg(-1) x min(-1) per pmol/l, r = 0.85 in meal; and DI: 1,282.26 vs. 1,273.23, r = 0.84 in OGTT and 726.92 vs. 776.97 10(-14) dl . kg(-1) x min(-2) per pmol/l, r = 0.84 in meal. This reduced protocol will facilitate the study of insulin secretion and action under physiological conditions in nondiabetic humans. PMID- 16249455 TI - Susceptibility to diabetic nephropathy is related to dicarbonyl and oxidative stress. AB - Dicarbonyl and oxidative stress may play important roles in the development of diabetes complications, and their response to hyperglycemia could determine individual susceptibility to diabetic nephropathy. This study examines the relationship of methylglyoxal, 3-deoxyglucosone (3DG), and oxidative stress levels to diabetic nephropathy risk in three populations with diabetes. All subjects in the Overt Nephropathy Progressor/Nonprogressor (ONPN) cohort (n = 14), the Natural History of Diabetic Nephropathy study (NHS) cohort (n = 110), and the Pima Indian cohort (n = 45) were evaluated for clinical nephropathy, while renal structural measures of fractional mesangial volume [Vv(Mes/glom)] and glomerular basement membrane (GBM) width were determined by electron microscopy morphometry in the NHS and Pima Indian cohorts. Methylglyoxal and 3DG levels reflected dicarbonyl stress, while reduced glutathione (GSH) and urine 8 isoprostane (8-IP) measured oxidative stress. Cross-sectional measures of methylglyoxal production by red blood cells incubated in 30 mmol/l glucose were increased in nephropathy progressors relative to nonprogressors in the ONPN (P = 0.027) and also reflected 5-year GBM thickening in the NHS cohort (P = 0.04). As nephropathy progressed in the NHS cohort, in vivo levels of methylglyoxal (P = 0.036), 3DG (P = 0.004), and oxidative stress (8-IP, P = 0.007 and GSH, P = 0.005) were seen, while increased methylglyoxal levels occurred as nephropathy progressed (P = 0.0016) in the type 2 Pima Indian cohort. Decreased glyceraldehyde-3-phosphate dehydrogenase activity also correlated with increased methylglyoxal levels (P = 0.003) in the NHS cohort. In conclusion, progression of diabetic nephropathy is significantly related to elevated dicarbonyl stress and possibly related to oxidative stress in three separate populations, suggesting that these factors play a role in determining individual susceptibility. PMID- 16249456 TI - Reduced nitric oxide concentration in the renal cortex of streptozotocin-induced diabetic rats: effects on renal oxygenation and microcirculation. AB - Nitric oxide (NO) regulates vascular tone and mitochondrial respiration. We investigated the hypothesis that there is reduced NO concentration in the renal cortex of diabetic rats that mediates reduced renal cortical blood perfusion and oxygen tension (P O2). Streptozotocin-induced diabetic and control rats were injected with l-arginine followed by Nomega-nitro-L-arginine-metyl-ester (L NAME). NO and P O2 were measured using microsensors, and local blood flow was recorded by laser-Doppler flowmetry. Plasma arginine and asymmetric dimethylarginine (ADMA) were analyzed by high-performance liquid chromatography. L-Arginine increased cortical NO concentrations more in diabetic animals, whereas changes in blood flow were similar. Cortical P O2 was unaffected by L-arginine in both groups. L-NAME decreased NO in control animals by 87 +/- 15 nmol/l compared with 45 +/- 7 nmol/l in diabetic animals. L-NAME decreased blood perfusion more in diabetic animals, but it only affected P O2 in control animals. Plasma arginine was significantly lower in diabetic animals (79.7 +/- 6.7 vs. 127.9 +/- 3.9 mmol/l), whereas ADMA was unchanged. A larger increase in renal cortical NO concentration after l-arginine injection, a smaller decrease in NO after L-NAME, and reduced plasma arginine suggest substrate limitation for NO formation in the renal cortex of diabetic animals. This demonstrates a new mechanism for diabetes induced alteration in renal oxygen metabolism and local blood flow regulation. PMID- 16249457 TI - Apoptotic stress is counterbalanced by survival elements preventing programmed cell death of dorsal root ganglions in subacute type 1 diabetic BB/Wor rats. AB - Several groups have reported apoptosis of dorsal root ganglion (DRG) cells as a prominent feature of diabetic polyneuropathy (DPN), although this has been controversial. Here, we examined subacute (4-month) type 1 diabetic BB/Wor rats with respect to sensory nerve functions, DRG and sural nerve morphometry, pro- and antiapoptotic proteins, and the expression of neurotrophic factors and their receptors. Sensory nerve conduction velocity was reduced by 13% and was accompanied by significant hyperalgesia. The numbers of DRG neurons including substance P-and calcitonin gene-related peptide-positive neurons were not altered, although they showed significant atrophy. Sural nerve morphometry showed decreased numbers of myelinated and unmyelinated fibers. Active caspase-3 and Bax expressions were increased, whereas antiapoptotic Bcl-xl and heat shock protein (HSP) 27 expressions in DRGs were increased. Nerve growth factor (NGF) contents in sciatic nerves and the expression of NGF receptor TrkA in DRGs were decreased. Immunohistochemistry showed increased numbers of active caspase-3-, HSP70-, and HSP27-positive neurons. Examinations of DRGs revealed no structural evidence of apoptosis but rather progressive hydropic degenerative changes. We conclude that apoptotic stress is induced in DRGs but is counterbalanced by survival elements in subacute type 1 diabetic BB/Wor rats and that distal nerve fiber loss reflects a dying-back phenomenon caused by impaired neurotrophic support. PMID- 16249458 TI - Protein tyrosine phosphatase-1B gene PTPN1: selection of tagging single nucleotide polymorphisms and association with body fat, insulin sensitivity, and the metabolic syndrome in a normal female population. AB - Protein tyrosine phosphatase-1B negatively regulates leptin and insulin signaling, potentially contributing to hormonal resistance. We selected six tagging single nucleotide polymorphisms (SNPs) representing 18 common variants in the protein tyrosine phosphatase-1B gene (PTPN1) and tested their effect on serum leptin, body fat, and measures of insulin sensitivity and the metabolic syndrome in a large sample of normal female Caucasian twins (n = 2,777; mean age, 47.4 +/- 12.5 years) from the St. Thomas' U.K. Adult Twin Registry. SNP rs718049 was significantly associated with waist circumference (P = 0.008) and central fat (P = 0.035) and also with Avignon's insulin sensitivity index (SiM) (P = 0.007), fasting insulin (P = 0.004), fasting glucose (P = 0.022), triglyceride (P = 0.023), and systolic blood pressure (P = 0.046). SNPs rs2282146 and rs1885177 were associated with SiM (P = 0.049 and P = 0.013, respectively), and 1484insG was associated with triglyceride (P = 0.029). A risk haplotype (7.3%) was associated with lower SiM (P = 0.036) and a protective haplotype (5.2%) with higher SiM (P = 0.057), with mean values in homozygotes differing by >1 SD (P = 0.003). The protective haplotype also showed lower triglyceride (P = 0.045) and lower systolic blood pressure (P = 0.006). Fine mapping analyses predicted significant associations with SiM and fasting insulin for several ungenotyped SNPs. PTPN1 variants appear to contribute to central fat and metabolic syndrome traits, secondary to their effect on insulin sensitivity. PMID- 16249459 TI - Assessment of 115 candidate genes for diabetic nephropathy by transmission/disequilibrium test. AB - Several lines of evidence, including familial aggregation, suggest that allelic variation contributes to risk of diabetic nephropathy. To assess the evidence for specific susceptibility genes, we used the transmission/disequilibrium test (TDT) to analyze 115 candidate genes for linkage and association with diabetic nephropathy. A comprehensive survey of this sort has not been undertaken before. Single nucleotide polymorphisms and simple tandem repeat polymorphisms located within 10 kb of the candidate genes were genotyped in a total of 72 type 1 diabetic families of European descent. All families had at least one offspring with diabetes and end-stage renal disease or proteinuria. As a consequence of the large number of statistical tests and modest P values, findings for some genes may be false-positives. Furthermore, the small sample size resulted in limited power, so the effects of some tested genes may not be detectable, even if they contribute to susceptibility. Nevertheless, nominally significant TDT results (P < 0.05) were obtained with polymorphisms in 20 genes, including 12 that have not been studied previously: aquaporin 1; B-cell leukemia/lymphoma 2 (bcl-2) proto oncogene; catalase; glutathione peroxidase 1; IGF1; laminin alpha 4; laminin, gamma 1; SMAD, mothers against DPP homolog 3; transforming growth factor, beta receptor II; transforming growth factor, beta receptor III; tissue inhibitor of metalloproteinase 3; and upstream transcription factor 1. In addition, our results provide modest support for a number of candidate genes previously studied by others. PMID- 16249461 TI - The fatty acid-binding protein-2 A54T polymorphism is associated with renal disease in patients with type 2 diabetes. AB - The intestinal fatty-acid binding protein-2 (FABP2) gene codes a protein responsible for the absorption of long-chain fatty acids. To test whether FABP2 is a candidate gene for renal disease in patients with type 2 diabetes, a functional A54T polymorphism was genotyped in 1,042 Brazilians with type 2 diabetes. Patients were classified as having normoalbuminuria (urinary albumin excretion [UAE] <20 microg/min; n = 529), microalbuminuria (UAE 20-199 microg/min; n = 217), or proteinuria (UAE >199 microg/min; n = 160). Patients with end-stage renal disease (ESRD) (n = 136) were also included. The prevalence of the TT genotype was higher in patients with renal involvement compared with those with normoalbuminuria (odds ratio [95% CI] 2.4 [1.1-5.4]) following adjustment for type 2 diabetes duration, BMI, hypertension, A1C, and cholesterol levels. The risk was similar considering different stages of renal involvement. In a second independent patient sample (483 type 2 diabetic Caucasians residing in Massachusetts), a significant association was also observed between the TT genotype and proteinuria or ESRD (2.7 [1.0-7.3]; P = 0.048). This study thus provides evidence that FABP2 confers susceptibility to renal disease in type 2 diabetic patients. PMID- 16249460 TI - Sequence variation in PPARG may underlie differential response to troglitazone. AB - Thiazolidinediones (TZDs) are peroxisome proliferator-activated receptor-gamma (PPARG) agonists used to treat type 2 diabetes. TZDs can also be used to reduce rates of type 2 diabetes in at-risk individuals. However, a large fraction of TZD treated patients (30-40%) do not respond to TZD treatment with an improvement in insulin sensitivity (Si). We hypothesized that variation within the gene encoding PPARG may underlie this differential response to TZD therapy. We screened approximately 40 kb of PPARG in 93 nondiabetic Hispanic women (63 responders and 30 nonresponders) with previous gestational diabetes who had participated in the Troglitazone In the Prevention Of Diabetes study. TZD nonresponse was defined as the lower tertile in change in Si after 3 months of treatment. Baseline demographic and clinical measures were not different between responders and nonresponders. We identified and genotyped 131 variants including 126 single nucleotide polymorphisms and 5 insertion-deletion polymorphisms. Linkage disequilibrium analysis identified five haplotype blocks. Eight variants were associated with TZD response (P < 0.05). Three variants were also associated with changes in Si as a continuous variable. Our results suggest that PPARG variation may underlie response to TZD therapy in women at risk for type 2 diabetes. PMID- 16249462 TI - Risk of diabetic nephropathy in type 1 diabetes is associated with functional polymorphisms in RANTES receptor gene (CCR5): a sex-specific effect. AB - Chemokines and their receptors have been implicated in the development of diabetic nephropathy. To determine whether the risk of diabetic nephropathy is influenced by two functional polymorphisms in the regulated upon activation normal T-cell expressed and secreted (RANTES) receptor gene (CCR5), we recruited patients with type 1 diabetes, including 496 case subjects with overt proteinuria or end-stage renal disease and 298 control subjects with normoalbuminuria. Male carriers of the 59029G allele, which is associated with diminished expression of CCR5 on the surface of immunocompetent cells, had significantly higher risk of developing diabetic nephropathy than noncarriers (OR [95% CI] 1.9 [1.2-3.0]). Similarly, male carriers of the 32-bp deletion, which causes truncation of the protein, had significantly higher risk of diabetic nephropathy than noncarriers (2.3 [1.3-4.2]). Combining both polymorphisms, three haplotypes were distinguished: one nonrisk haplotype carrying the 59029A allele and the 32-bp insertion and two risk haplotypes carrying the 59029A allele with the 32-bp deletion and carrying the 59029G allele with the 32-bp insertion. The distribution of these haplotypes differed significantly (P < 0.00001) in men with and without diabetic nephropathy but was not associated with diabetic nephropathy in women. In conclusion, two functional polymorphisms in CCR5 that decrease expression of the RANTES receptor on immunocompetent cells are associated with increased risk of diabetic nephropathy in type 1 diabetes, but only in men. PMID- 16249463 TI - Altered calcium homeostasis is correlated with abnormalities of fasting serum glucose, insulin resistance, and beta-cell function in the Newfoundland population. AB - Alteration of extracellular calcium concentration may be involved in glucose metabolism in a number of pathways. The present study was designed to investigate the relationship between total serum calcium and 1) fasting serum glucose, 2) insulin, 3) insulin resistance, and 4) beta-cell function in 1,182 healthy subjects from the province of Newfoundland and Labrador, Canada. All variables were log10 transformed, and confounding factors including age, trunk fat percentage, serum phosphorus, magnesium, 25-OH vitamin D, and parathyroid hormone were adjusted before analyses. Significant positive correlations between glucose and insulin resistance with calcium were found in both sexes, whereas an inverse correlation between beta-cell function and calcium was found only in women. Similar results were found in medication-free women and men, as well as in pre- and postmenopausal women. Subjects with low calcium levels had the lowest concentration of glucose and the least insulin resistance, whereas subjects with high calcium levels had the highest concentration of glucose and insulin resistance in women but not in men. This relationship remained after calcium was adjusted for 25-OH vitamin D and parathyroid hormone. Our results suggest that alteration of serum calcium homeostasis is significantly correlated with the abnormality of glucose level, insulin resistance, and beta-cell function. PMID- 16249464 TI - Autologous T-lymphocytes stimulate proliferation of orbital fibroblasts derived from patients with Graves' ophthalmopathy. AB - PURPOSE: Graves' ophthalmopathy (GO) affects 50% to 60% of patients with Graves' hyperthyroidism, resulting in exophthalmos, periorbital edema, pain, double vision, optic neuropathy, and loss of vision. Fibroblasts are a key autoimmune target in GO and have effector functions that contribute to GO-associated pathologic conditions, including proliferation, production of excess glycosaminoglycans, and fat deposition. GO is also characterized by autoimmune inflammation of orbital connective tissue with mononuclear cell infiltration, including T cells. METHODS: To determine whether autologous T cells can drive proliferation of orbital fibroblasts and thus contribute to GO, a novel reverse autologous mixed-cell reaction (rAMCR) was performed. Fibroblasts cultured from orbital tissue of patients with GO that was removed during orbital decompression surgery were mixed with autologous T cells, and fibroblast proliferation was determined. RESULTS: Autologous T cells stimulated proliferation of orbital fibroblasts. Fibroblasts derived from blepharoplasty fat of two different patients did not proliferate, demonstrating that the effect is specific to cells derived from deep orbital fat. Proliferation was dependent on cell contact and on major histocompatibility complex (MHC) class II and CD40-CD154 (CD40 ligand) signaling. CONCLUSIONS: The results suggest that T cells and orbital fibroblasts participate in an antigen-dependent positive feedback loop in which presentation of autoantigens by fibroblasts via MHC class II and CD40-CD40L signaling results in T-cell activation. These activated T cells stimulate fibroblast proliferation, leading to fibroblast-associated diseases in GO. Thus, therapies that interfere with CD40-CD40L signaling, antigen expression by fibroblasts, or T-cell function may be effective in preventing progression of GO symptoms. PMID- 16249465 TI - Glucagon receptor agonists and antagonists affect the growth of the chick eye: a role for glucagonergic regulation of emmetropization? AB - PURPOSE: In chicks, plus defocus retards eye growth, thickens the choroid, and activates glucagonergic amacrine cells, probably releasing glucagon. Glucagon receptor antagonists (expected to inhibit compensation to plus defocus) and agonists (expected to block myopia induction by form deprivation) were administered to eyes of chicks, to test the hypothesis that glucagon mediates the induction of changes in eye growth by plus defocus. METHODS: Seven-day-old (P7) chick eyes were injected intravitreally with peptides at concentrations of approximately 10(-9) to 10(-5) M in 20 microL (injection volume). The glucagon receptor antagonists [des-His(1),des- Phe(6),Glu(9)]-glucagon-NH(2) (des- Phe(6) antagonist) and [des-His(1),Glu(9)]-glucagon-NH(2) (Phe(6)-antagonist) were administered daily for 4 to 5 days to plus-defocused eyes. Agonists (porcine glucagon-[1-29] and [Lys(17,18),Glu(21)]-glucagon-NH(2)) were monocularly administered daily for 5 days to form-deprived eyes. The contralateral eye remained open and received saline. After treatment, eyes were refracted, measured, and examined for histologic changes. RESULTS: The Phe(6)-antagonist at 10(-5) M (in the syringe) inhibited changes in both refractive error and axial length compensation induced by +7-D lens wear; however, des-Phe(6)-antagonist (10(-5) M) had weak, inconsistent effects and did not antagonize the action of exogenous glucagon. Glucagon prevented ocular elongation and myopia and induced choroidal thickening in form-deprived eyes. [Lys(17,18),Glu(21)]-glucagon-NH(2) had little effect at 10(-7) M, but at 10(-6) to 10(-5) M altered rod structure and inhibited eye growth. CONCLUSIONS: Exogenous glucagon inhibited the growth of form-deprived eyes, whereas Phe(6)-antagonist inhibited compensation to plus defocus, as might be expected if glucagon is an endogenous mediator of emmetropization. The reason for the failure of des-Phe(6)-antagonist to counteract the effects of exogenous glucagon requires further investigation. PMID- 16249466 TI - Glucagon- and secretin-related peptides differentially alter ocular growth and the development of form-deprivation myopia in chicks. AB - PURPOSE: Exogenous glucagon inhibits the induction of myopia in chicks, but the endogenous peptide and receptor that regulate eye growth are unknown. The purpose of this study was to determine which peptides and receptors in the glucagon secretin family play a role in the control of ocular growth. METHODS: The effect of intravitreally injected peptides on the development of form-deprivation (FD) myopia and on the growth of eyes with unrestricted vision was determined by refraction and A-scan ultrasonography. Chicks received three injections, one every 48 hours, of secretin-related peptides (porcine secretin, human peptide histidine-isoleucine-amide-27, vasoactive intestinal peptide [VIP], VIP fragment 6-28, or pituitary adenylate cyclase-activating polypeptide; 10(-8)-10(-4) M in 20 microL) or five injections of proglucagon-derived peptides (human glucagon, oxyntomodulin, miniglucagon, or glucagon-like peptide [GLP]-2 or chicken GLP-1). Immunohistochemistry was used to detect proglucagon-derived peptides in the eye. RESULTS: Secretin-related peptides had no effect on FD myopia, whereas some proglucagon-derived peptides did. Both glucagon and oxyntomodulin dose dependently inhibited development of myopia, primarily by inhibition of vitreous chamber elongation (EC(50) = 10(-4) M and 10(-5.5) M, respectively). GLP-1 increased deprivation-induced myopic refractive error by altering anterior chamber development. None of the peptides significantly affected refractive error in eyes with unrestricted vision, although changes in anterior and posterior eye growth were observed in response to glucagon, oxyntomodulin, GLP-1, and miniglucagon. Immunoreactivity for miniglucagon and GLP-1 was colocalized in glucagon-immunoreactive amacrine cells. CONCLUSIONS: Prevention of experimental myopia by exogenous glucagon is mediated by receptors selective for glucagon and oxyntomodulin, indicating that glucagon-like peptides and receptors are probable endogenous retinal regulators of the development of myopia. PMID- 16249467 TI - Idiopathic bilateral optic atrophy in the rhesus macaque. AB - PURPOSE: To document the existence of idiopathic bilateral optic atrophy (BOA) in rhesus macaque monkeys and to characterize the structural and functional consequences of this condition. METHODS: In vivo assessment of retinal and optic nerve structure included fundus biomicroscopy and stereophotography. Functional analyses included transient pattern-reversal electroretinography (PERG) and full field flash ERG, with both white flashes while dark adapted and red flashes on a blue background used to assess the photopic negative response (PhNR). Also measured were visual evoked cortical potentials (VEPs) and multifocal (mf)ERGs, with both a standard fast and slowed (7F) stimulation sequence. Post mortem histologic evaluation was performed on a subset of five animals with BOA and compared with data from 22 healthy normal animals. Blood tests, including vitamin E, B(12), folate, lead, and complete blood cell count with differential were obtained on the four animals that remained alive. RESULTS: Animals with BOA showed temporal pallor of the optic nerve head and thinning of the retinal nerve fiber layer (RNFL) between the temporal vascular arcades (i.e., of the papillomacular bundle). Severity of optic atrophy and RNFL loss varied between animals from mild to severe, but was similar in the two eyes of each animal. Functional changes included greater loss of the PERG N95, compared with the P50 component and substantial reduction of mfERG high-frequency components. The mfERG low-frequency components were slightly larger than normal. None of the full-field flash ERG amplitudes (a-wave, b-wave, oscillatory potentials, or PhNR) was significantly different from normal. There were no consistent abnormalities found in the results of any blood test. Histologic findings included axonal loss and gliosis limited to the temporal optic nerve, reduction of nuclei within the retinal ganglion cell layer, and thinning of the temporal retinal RNFL. CONCLUSIONS: The existence of BOA in nonhuman primates warrants caution on the part of investigators who use these animals in experimental models of ophthalmic disease. PMID- 16249468 TI - Expression of immunoglobulin transcription factors in primary intraocular lymphoma and primary central nervous system lymphoma. AB - PURPOSE: Several B-cell-associated transcription factors and their coactivators, including BCL-6, BSAP/PAX5, BOB.1/OBF.1, Oct.2, MUM1/IRF4, and PU.1, have been detected in peripheral B-cell lymphomas. There are limited data on their expression in centrally located lymphoid neoplasms, such as primary intraocular lymphoma (PIOL) or primary central nervous system lymphoma (PCNSL). PIOL is a rare non-Hodgkin lymphoma, considered a subtype of PCNSL. Both are usually diffuse, large B-cell lymphoma (DLBCL), rarely manifest outside the CNS, and carry a poor prognosis. METHODS: Tissue biopsy specimens were examined from eight cases of PIOL and 42 cases of HIV-negative PCNSL, as well as 50 cases of peripheral DLBCL, for the above-mentioned transcription factors and for immunoglobulin heavy and light chains, using immunohistochemistry. RESULTS: Immunoglobulin expression was demonstrated in 46 (92%) of 50 cases of PIOL/PCNSL but in only 27 (54%) of 50 cases of peripheral DLBCL. Positivity for BOB.1/OBF.1 and Oct.2 was observed in all immunoglobulin-expressing PIOL and PCNSL. BSAP/PAX5 expression occurred in 98% of PIOL/PCNSL, and MUM1/IRF4 immunoreactivity in 45 (90%) of 50 of these cases. PU.1 expression was observed in only 10% of the PIOL/PCNSL group in contrast to 23 (46%) of 50 peripheral DLBCLs. Aberrant coexpression of MUM1/IRF4, PAX5, MUM1/IRF4, and BCL-6 was observed in most PIOLs/PCNSLs. CONCLUSIONS: These data provide further support to the notion that peripheral and centrally located DLBCLs differ in clinical, immunophenotypic, and genotypic features, despite their similar morphologic characteristics. PIOL and PCNSL tumor cells are most likely to be derived from mature B-cells that have undergone the germinal center reaction. PMID- 16249470 TI - Aberrant accumulation of fibulin-3 in the endoplasmic reticulum leads to activation of the unfolded protein response and VEGF expression. AB - PURPOSE: The inherited early-onset macular degenerative disease known as malattia leventinese (ML) and Doyne honeycomb retinal dystrophy (DHRD) have been linked to a missense mutation leading to production of a mutant fibulin-3 protein (R345W). R345W is poorly secreted and accumulates in the RPE of ML/DHRD retinas. Accumulation of misfolded proteins within the endoplasmic reticulum (ER) causes activation of unfolded protein response (UPR) signaling and expression of ER stress-responsive genes, including vascular endothelial growth factor (VEGF). Therefore, we hypothesized that the expression of R345W activates the UPR, leading to VEGF expression. METHODS: Adenoviral vectors were used to overexpress fibulin-3 wild-type (Wt) and R345W mutant proteins in ARPE-19 cells. Secretion and intracellular accumulation of Wt and R345W were compared by Western blot analysis and immunocytochemistry. Activation of the UPR was evaluated by measuring the expression of glucose-regulated protein 78 (GRP78 [BiP]) and editing of the X-box binding protein (XBP-1) mRNA. VEGF expression and transcriptional activation of the VEGF promoter were determined by Northern blot analysis, Western blot analysis, and use of a novel VEGF promoter-reporter construct containing 8.2 kb of the human VEGF gene. RESULTS: R345W was poorly secreted by ARPE-19 cells and accumulated in the ER, leading to UPR activation and increased VEGF expression. Compared with Wt mutant proteins, the expression of R345W was more effective at causing UPR activation, increasing VEGF expression, and stimulating transcription from the VEGF promoter. CONCLUSIONS: These findings demonstrated that the expression of mutated fibulin-3 caused UPR activation and increased VEGF expression. Expression of mutant fibulin proteins may contribute to macular degeneration and choroidal neovascularization by causing ER stress leading to RPE dysfunction and increased VEGF expression. PMID- 16249469 TI - Peripheral vision can influence eye growth and refractive development in infant monkeys. AB - PURPOSE: Given the prominence of central vision in humans, it has been assumed that visual signals from the fovea dominate emmetropization. The purpose of this study was to examine the impact of peripheral vision on emmetropization. METHODS: Bilateral, peripheral form deprivation was produced in 12 infant monkeys by rearing them with diffusers that had either 4- or 8-mm apertures centered on the pupils of each eye, to allow 24 degrees or 37 degrees of unrestricted central vision, respectively. At the end of the lens-rearing period, an argon laser was used to ablate the fovea in one eye of each of seven monkeys. Subsequently, all the animals were allowed unrestricted vision. Refractive error and axial dimensions were measured along the pupillary axis by retinoscopy and A-scan ultrasonography, respectively. Control data were obtained from 21 normal monkeys and 3 infants reared with binocular plano lenses. RESULTS: Nine of the 12 treated monkeys had refractive errors that fell outside the 10th- and 90th-percentile limits for the age-matched control subjects, and the average refractive error for the treated animals was more variable and significantly less hyperopic/more myopic (+0.03 +/- 2.39 D vs. +2.39 +/- 0.92 D). The refractive changes were symmetric in the two eyes of a given animal and axial in nature. After lens removal, all the treated monkeys recovered from the induced refractive errors. No interocular differences in the recovery process were observed in the animals with monocular foveal lesions. CONCLUSIONS: On the one hand, the peripheral retina can contribute to emmetropizing responses and to ametropias produced by an abnormal visual experience. On the other hand, unrestricted central vision is not sufficient to ensure normal refractive development, and the fovea is not essential for emmetropizing responses. PMID- 16249471 TI - Expression and function of glutamine transporters SN1 (SNAT3) and SN2 (SNAT5) in retinal Muller cells. AB - PURPOSE: The expression and function of the glutamine transporters ATA1 and ATA2 (isoforms of system A), SN1 and SN2 (isoforms of system N), and LAT1 and LAT2 (isoforms of system L) were investigated in Muller cells in a rat Muller cell line (rMC1) and primary cultures of mouse Muller cells. METHODS: Glutamine uptake in rMC1 cells and primary Muller cells was measured. The relative contributions of various transport systems to glutamine uptake were determined based on the differential substrate specificities and Na(+) dependence of individual transport systems. RT-PCR was used to analyze the expression of transporter-specific mRNAs. RESULTS: Three different transport systems participated in glutamine uptake in rMC1 cells: system L (Na(+)-independent), system A (Na(+)-dependent and alpha (methylamino)isobutyric acid [MeAIB]-sensitive), and system N (Na(+)-dependent and MeAIB-insensitive). System N was the principal contributor (approximately 70%); the contributions of systems A and L were relatively lesser (20% and <10%, respectively). The functional features of Na(+)-dependent and MeAIB-insensitive glutamine uptake were similar to the known characteristics of clones of SN1 and SN2. Glutamine uptake in primary Muller cells behaved in a manner similar to that in rMC1 cells. mRNA transcripts specific for ATA1, ATA2, SN1, SN2, LAT1, and LAT2 were expressed in Muller cells. CONCLUSIONS: System N (SN1 as well as SN2) is responsible for most of the glutamine uptake in Muller cells. Because system N is capable of mediating the release of glutamine from the cells, its abundant expression in Muller cells is of importance in the handling of glutamine in the retina. PMID- 16249473 TI - Objective quantification of posterior capsule opacification after cataract surgery, with optical coherence tomography. AB - PURPOSE: To evaluate posterior capsule opacification (PCO) in humans after cataract surgery with intraocular lens (IOL) implantation, by using optical coherence tomography (OCT-1). METHODS: Sixty-six eyes with PCO and 20 eyes with a normal posterior capsule were analyzed. A 3-mm-long horizontal scan of the posterior capsule was obtained. Measurements at three points and their average were recorded. Intraoperator and interoperator reliabilities were assessed. Investigated was peak intensity (PI) and posterior capsule thickening (PCT), with PCT indicating the distance between two reflectivity spikes, with an approximate axial resolution of 10 microm. Results were compared with visual acuity (VA) and PCO type. RESULTS: Intraoperator reliability was 0.59 and 0.97 for average PI and PCT, respectively. The interoperator concordance correlation coefficient was 0.70 and 0.82 for average PI and PCT, respectively. Median (interquartile range) intensities of the reflectivity spike were 16.88 (dB) (range, 12.88-20.41) and 11.9 (8.58-14.28), respectively, in the PCO and control eyes (P = 0.001). PCT was found in PCO eyes (median: 86.13 microm; range, 46.33-115.33), whereas no second spike appeared in control eyes (P = 0.001). The area under the receiver operating characteristic curve of the average PCT for differentiating pearl-type from fibrosis-type PCO was 0.87 (P = 0.001). For a cutoff point of 55.3 microm, the sensitivity was 97.5%, and the specificity was 69%. Worse VA correlated significantly only with larger PCT (r(o) = 0.66; P = 0.01). CONCLUSIONS: OCT-1 appears useful to quantitate PCO. In addition, this system seems to discriminate between different types of PCO. PCT may be a previously unrecognized factor in VA degradation. PMID- 16249472 TI - A role for cytoskeletal elements in the light-driven translocation of proteins in rod photoreceptors. AB - PURPOSE: Light-driven protein translocation is responsible for the dramatic redistribution of some proteins in vertebrate rod photoreceptors. In this study, the involvement of microtubules and microfilaments in the light-driven translocation of arrestin and transducin was investigated. METHODS: Pharmacologic reagents were applied to native and transgenic Xenopus tadpoles, to disrupt the microtubules (thiabendazole) and microfilaments (cytochalasin D and latrunculin B) of the rod photoreceptors. Quantitative confocal imaging was used to assess the impact of these treatments on arrestin and transducin translocation. A series of transgenic tadpoles expressing arrestin truncations were also created to identify portions of arrestin that enable arrestin to translocate. RESULTS: Application of cytochalasin D or latrunculin B to disrupt the microfilament organization selectively slowed only transducin movement from the inner to the outer segments. Perturbation of the microtubule cytoskeleton with thiabendazole slowed the translocation of both arrestin and transducin, but only in moving from the outer to the inner segments. Transgenic Xenopus expressing fusions of green fluorescent protein (GFP) with portions of arrestin implicates the C terminus of arrestin as an important portion of the molecule for promoting translocation. This C-terminal region can be used independently to promote translocation of GFP in response to light. CONCLUSIONS: The results show that disruption of the cytoskeletal network in rod photoreceptors has specific effects on the translocation of arrestin and transducin. These effects suggest that the light driven translocation of visual proteins at least partially relies on an active motor-driven mechanism for complete movement of arrestin and transducin. PMID- 16249474 TI - Vision and quality of life: the development of a utility measure. AB - PURPOSE: To identify the content for a vision and quality of life-related utility measure (Vision Quality of Life Index [VisQoL]) for the economic evaluation of eye care and rehabilitation programs. METHODS: Focus groups of the visually impaired elicited key concepts. Based on these and previous research, 33 items were generated. These were administered to visually impaired adults (n = 70) and a representative sample of unimpaired adults (n = 86). The item bank was reduced through examination of item properties, exploratory factor (EFA), item response theory (IRT), and structural equation modeling (SEM) analyses. The resultant model was confirmed through administration to a second sample of participants. RESULTS: Focus group themes included physical well-being, social well-being, independence, self-actualization, emotional well-being, and planning and organization. Poorly performing items were eliminated on basic psychometric properties, including failure to discriminate. Next, EFA loadings were used to select items. Twelve items survived. To minimize redundancy, IRT analysis and SEM reduced the VisQoL item pool to six items (Cronbach alpha = 0.88). To confirm this model, these items were then administered to an additional 218 participants; 35% with a vision impairment. A pooled SEM analysis showed the model to have very good fit properties (root mean square error of approximation [RMSEA] = 0.000). A preliminary test of the model against visual acuity showed a significant monotonic relationship. CONCLUSIONS: The short 6-item VisQoL has excellent psychometric properties as a simple summative instrument. It can be used in its present state as a condition-specific outcome measure for the evaluation of healthcare interventions for the visually impaired. The descriptive model is also suitable for generating utility values for the economic evaluation of vision related programs and services. PMID- 16249475 TI - The impact of age-related macular degeneration on health status utility values. AB - PURPOSE: To estimate health status utility values in patients with age-related macular degeneration (ARMD) associated with visual impairments, by using preference-based measures of health. METHOD: This was a cross-sectional study involving patients with unilateral or bilateral ARMD who attended a large teaching hospital. Patients underwent visual tests (near and distant visual acuity [VA] and contrast sensitivity [CS]) and completed health status questionnaires including the Index of Visual Function (VF)-14 and three preference-based measures (the Health Utilities Index Mark III [HUI-3], the EuroQoL Health Questionnaire [EQ-5D], and the Short Form 6D Health Status Questionnaire [SF-6D]) and the time tradeoff (TTO). The mean health status is presented for five groups, defined according to the VA in the better-seeing eye and for four CS groups. RESULTS: Two hundred nine patients were recruited with substantial loss of visual function as obtained by visual tests (mean decimal VA in the better-seeing eye: 0.2) and self-report (mean VF-14 score: 41.5). The mean (+/-SD) utilities were 0.34 +/- 0.28 for HUI-3, 0.66 +/- 0.14 for SF-6D, 0.72 +/- 0.22 for EQ-5D, and 0.64 +/- 0.31 for TTO. The HUI-3 had the highest correlation with VA and CS (0.40 and -0.34), followed by TTO (0.25 and -0.21). Across the VA and CS groups, only HUI3 and TTO had a significant linear trend (P < 0.05). In a regression model with CS and VA as explanatory variables, only the coefficient on CS was statistically significant. CONCLUSIONS: ARMD is associated with a substantial impact on patients' health status, but this was not reflected in two of the generic preference-based measures used. The HUI-3 seems to be the instrument of choice for use in economic evaluations in which community data are needed. It may be more appropriate to base economic models on CS or some combination of CS and VA rather than on VA alone. PMID- 16249476 TI - Anisometropia is independently associated with both spherical and cylindrical ametropia. AB - PURPOSE: To explore the associations between anisometropia and spherical ametropia, astigmatism, age, and sex. METHOD: Associations between the prevalence and magnitude of anisometropia with age, sex, spherical power, and cylindrical power, were assessed in a group of 90,884 subjects attending optometry practices in the United Kingdom. Logistic regression models were used to assess the independent contribution of each explanatory variable. RESULTS: Logistic regression analyses that included all subjects or just those aged 20 to 40 years showed that spherical ametropia and astigmatism were independently associated with anisometropia (myopes, P < 1.0E-61; hyperopes, P < 1.0E-11). Anisometropia was relatively stable between the ages of 20 and 40 years, but then became more common with age, in myopes from the age of 40 years onward (P < 0.003) and in hyperopes from the age of 70 years onward (P < 1.0E-6). Sex was not associated with anisometropia to a clinically significant extent. CONCLUSIONS: This is the first study to show an independent association between anisometropia and both spherical ametropia and astigmatism. The results also suggest that the previously noted increased prevalence of anisometropia with age occurs later in hyperopes than in myopes, once other covariates have been controlled for. However, it could not be ruled out that this latter effect was due to clinical selection bias in our sample. The findings suggest that research projects involving the recruitment of highly ametropic subjects, such as those investigating the genetics of refractive error, may benefit by avoiding the use of stringent inclusion criteria for anisometropia, because otherwise a large proportion of the relevant population will be excluded. PMID- 16249477 TI - Nine-year refractive changes in the Barbados Eye Studies. AB - PURPOSE: To describe 9-year changes in refractive errors and estimate incidence of myopia and hyperopia in adults of African-descent, along with associated risk factors. METHODS: The 9-year follow-up of the Barbados Eye Studies (1997-2003) reexamined 2793 surviving cohort members (81% participation). Refractive errors were determined by automated refraction. Myopia/hyperopia were defined as spherical equivalent < -0.5 diopters (D)/> +0.5 D, and the cutoff for moderate high myopia/hyperopia was 3.0 D. Incidence rates of myopia/hyperopia were estimated by the product-limit approach, based on eyes without such conditions at baseline. Risk factors were evaluated by logistic regression in discrete time hazard models. RESULTS: Nine-year refraction changes varied by age. Persons aged 40 to 49 years experienced hyperopic shifts (median, +0.38 D), whereas persons > or =60 years had myopic shifts (median, -0.75D). Overall 9-year incidence was 12.0% for myopia and 29.5% for hyperopia; rates were 3.6% and 2.0% for moderate high myopia and hyperopia, respectively. Myopia risk increased with age, baseline nuclear lens opacities (risk ratio [RR] = 1.7; 95% confidence interval [CI]: 1.01 2.9), glaucoma (RR = 6.0, 95% CI: 3.9-9.3), and ocular hypertension (RR = 2.0, 95% CI: 1.3-3.0), while cortical lens opacities decreased risk (RR = 0.6, 95% CI: 0.4-0.9). Incidence of moderate-high myopia was also related to baseline age, nuclear opacities, glaucoma, male gender (RR = 1.7, 95% CI: 1.0-2.8), and diabetes history (RR = 1.9, 95% CI: 1.01-3.5). Hyperopia risk decreased with older age, male gender, and glaucoma diagnosis. CONCLUSIONS: Refractive errors continue to develop frequently in older adults. Nuclear lens opacities, glaucoma, and diabetes increase the risk of older-onset myopia, a result of public health relevance to this and similar African-origin populations. PMID- 16249478 TI - Visual acuity change and mortality in older adults. AB - PURPOSE: Several studies indicate an increased mortality rate in older adults who have visual impairment, but few have attempted to address a potential causal mechanism. The goals of this study are to determine whether visual acuity loss increases the risk of dying and to examine whether depressive symptoms act as a mediator in this relationship. METHODS: Data were derived from the 2520 older adults who participated in the Salisbury Eye Evaluation project, a population based prospective 8-year cohort study. Presenting binocular visual acuity was measured with the Early Treatment Diabetic Retinopathy Study [ETDRS] eye chart and depressive symptoms with the General Health Questionnaire Part D subscale. Mortality data were collected by staff follow-up. Analyses were performed with the Cox proportional hazards regression. RESULTS: Worse baseline acuity was associated with a higher mortality rate (hazard ratio [HR] = 1.05; 95% confidence interval [CI], 1.01-1.09). Also, those who gained two or more lines of visual acuity over 2 years had a lower adjusted risk of dying (HR = 0.47; 95% CI, 0.23 0.95). An interaction was detected, in that women who lost > or =3 lines of visual acuity over a 2-year period had a higher adjusted risk of dying (HR = 3.97; 95% CI, 2.21-7.15), whereas men did not (HR = 1.32; 95% CI, 0.66-2.63). Depressive symptoms did not mediate these relationships. CONCLUSIONS: If the relationship between visual acuity and mortality is indeed causal, it most likely acts via numerous pathways through a variety of intervening variables. The identification of these intervening variables could give additional targets for intervention if acuity cannot be restored. PMID- 16249479 TI - An x-ray diffraction study of corneal structure in mimecan-deficient mice. AB - PURPOSE: Keratan sulfate proteoglycans (KSPGs) in the corneal stroma are believed to influence collagen fibrillar arrangement. This study was performed to investigate the fibrillar architecture of the corneal stroma in mice homozygous for a null mutation in the corneal KSPG, mimecan. METHODS: Wild-type (n = 9) and mimecan-deficient (n = 10) mouse corneas were investigated by low-angle synchrotron x-ray diffraction to establish the average collagen fibrillar spacing, average collagen fibril diameter, and level of fibrillar organization in the stromal array. RESULTS: The mean collagen fibril diameter in the corneas of mimecan-null mice, as an average throughout the whole thickness of the tissue, was not appreciably different from normal (35.6 +/- 1.1 nm vs. 35.9 +/- 1.0 nm). Average center-to-center collagen fibrillar spacing in the mutant corneas measured 52.6 +/- 2.6 nm, similar to the 53.3 +/- 4.0 nm found in wild-type mice. The degree of local order in the collagen fibrillar array, as indicated by the height-width (H:W) ratio of the background-subtracted interfibrillar x-ray reflection, was also not significantly changed in mimecan-null corneas (23.4 +/- 5.6), when compared with the corneas of wild-types (28.2 +/- 4.8). CONCLUSIONS: On average, throughout the whole depth of the corneal stroma, collagen fibrils in mimecan-null mice, unlike collagen fibrils in lumican-null mice and keratocan null mice, are of a normal diameter and are normally spaced and arranged. This indicates that, compared with lumican and keratocan, mimecan has a lesser role in the control of stromal architecture in mouse cornea. PMID- 16249481 TI - Plasminogen kringle 5 inhibits alkali-burn-induced corneal neovascularization. AB - PURPOSE: Plasminogen kringle 5 (K5) is a potent angiogenic inhibitor. The purpose of the present study was to evaluate the therapeutic effect of K5 on alkali-burn induced corneal neovascularization (NV) and to investigate its mechanism of action. METHODS: Corneal NV was induced in rabbits by NaOH. The rabbits received eye drops containing K5 or vehicle alone, four times per day. Corneal NV and inflammation were monitored every other day with a slit lamp microscope, and the length of the vessels in the cornea and the area of NV were measured. Vascular endothelial growth factor (VEGF) was determined by immunohistochemical and Western blot analyses. The TUNEL assay was used to assess the apoptosis of endothelial cells. The effects of K5 on primary bovine aortic endothelial cells (BAECs) were determined by MTT assay, flow cytometry, transmission electron microscopy, and DNA fragmentation assay. RESULTS: Alkali-burn-induced progressive corneal NV and inflammation in the cornea. K5 delayed the onset of corneal NV (P < 0.05) and decreased NV areas (P < 0.05) in a dose-dependent manner. K5 treatment, after the formation of corneal NV, induced regression of newly formatted vessels in the cornea. K5 decreased the inflammatory index in the corneas at different time points after the alkali burn. Corneal VEGF levels were reduced by K5 treatment. K5 inhibits proliferation and induces apoptosis in BAECs. CONCLUSIONS: Topical application of K5 may have therapeutic potential for the chemical burn-induced corneal NV and inflammation. The inhibitory effect of K5 on corneal NV may be by downregulation of VEGF expression. PMID- 16249482 TI - Downregulation of IRS-1 expression causes inhibition of corneal angiogenesis. AB - PURPOSE: The antiangiogenic effect of an antisense oligodeoxynucleotide (ODN) targeting insulin receptor substrate (IRS)-1 was evaluated on rat corneal neovascularization. METHODS: Eyes with neovessels were treated with subconjunctival injections of IRS-1 antisense oligonucleotide (ASODN), IRS-1 sense ODN (SODN), or PBS. At 8 and 24 hours after the first subconjunctival injection, the expression of IRS-1, VEGF, and IL-1beta mRNA was evaluated. IRS-1 protein levels were also measured at 8 hours by Western blot analysis (n = 4/group). On day 10, corneal neovascularization was quantified in flatmount corneas of rats treated daily from days 4 to 9. RESULTS: On day 10, new vessels covered 95.5% +/- 4% of the corneal area in PBS-treated eyes, 92% +/- 7% in SODN treated eyes and 59% +/- 20% in ASODN-treated eyes (P < 0.001). In the ASODN treated group, the expression and synthesis of IRS-1 were significantly downregulated when compared with the control groups. ASODN did not significantly affect the expression of VEGF but significantly decreased the expression of IL 1beta at 24 hours (P = 0.04). CONCLUSIONS: Subconjunctival injections of IRS-1 antisense ODN significantly inhibit rat corneal neovascularization. This effect may be mediated by a downregulation of IL-1beta. IRS-1 proteins may be interesting targets for the regulation of angiogenesis mediated by insulin, hypoxia, or inflammation. PMID- 16249480 TI - Effect of retinoic acid on gene expression in human conjunctival epithelium: secretory phospholipase A2 mediates retinoic acid induction of MUC16. AB - PURPOSE: How vitamin A contributes to the maintenance of the wet-surfaced phenotype at the ocular surface is not well understood. This study sought to identify vitamin A-responsive genes in ocular surface epithelia using gene microarray analysis of cultures of a human conjunctival epithelial (HCjE) cell line grown with all-trans-retinoic acid (RA). The analysis showed that secretory phospholipase A(2) group IIA (sPLA(2)-IIA) was the gene most upregulated by RA, followed by the membrane-associated mucin MUC16 at a later time point. Since eicosanoids, the product of arachidonic acid generated by the PLA(2) family, have been shown to increase mucin production, this study sought to determine whether sPLA(2) mediates the RA induction of MUC16. METHODS: HCjE cells were cultured with or without RA for 3, 6, 24, and 48 hours. Complementary RNA prepared from RNA of the HCjE cells was hybridized to human gene chips and analyzed using commercial software. Microarray data on mucin expression were validated by real time PCR. To investigate whether sPLA(2) is associated with RA-induced MUC16 upregulation, HCjE cells were incubated with RA and the broad-spectrum PLA(2) inhibitor aristolochic acid (ArA) or the specific sPLA(2)-IIA inhibitor LY315920, followed by analysis of MUC16 mRNA and protein by real-time PCR and Western blot analysis. RESULTS: After RA addition, 28 transcripts were upregulated and 6 downregulated by more than twofold (P < 0.01) at both 3 and 6 hours (early phase). Eighty gene transcripts were upregulated and 45 downregulated at both 24 and 48 hours (late phase). Group IIA sPLA(2), significantly upregulated by 24 hours, and MUC16 were the most upregulated RNAs by RA at 48 hours. sPLA(2) upregulation by RA was confirmed by Western blot analysis. When HCjE cells were incubated with RA plus ArA or specific inhibitor of sPLA(2)-IIA, LY315920, the RA induced MUC16 mRNA was significantly reduced (P < 0.01). CONCLUSIONS: The RA associated upregulation of membrane-associated mucin MUC16 at late phase appears to be through sPLA(2)-IIA. Upregulation of this hydrophilic membrane-associated mucin may be one of the important mechanisms by which vitamin A facilitates maintenance of the wet-surfaced phenotype on the ocular surface. PMID- 16249483 TI - Transplantation of amniotic membrane in murine herpes stromal keratitis modulates matrix metalloproteinases in the cornea. AB - PURPOSE: To study matrix metalloproteinases (MMP) and tissue inhibitors of metalloproteinases (TIMP) in the corneas from mice with ulcerative herpes stromal keratitis (HSK) treated with amniotic membrane transplantation (AMT). METHODS: The corneas from BALB/c mice were infected with HSV-1. Mice with ulcerative HSK on postinfection (PI) day 14 were used for the experiments. In one group of mice, the corneas were treated with transplantation of amniotic membrane (AMT) that was secured with a tarsorrhaphy, and a control group underwent tarsorrhaphy alone. After 2 days, the appearance of corneal ulcers and stromal inflammation was judged clinically. Corneal sections were studied by immunohistochemistry for the expression of MMP-2, -8, and -9 and TIMP-1 and -2. MMP activity in the corneas was investigated by zymography, and the expression of the enzymes was measured by the Western blot technique. RESULTS: At day 14 PI, the ulcers stained intensely positive for MMP-2, -8, and -9 and TIMP-1 and -2. Ulceration (P < 0.001), stromal inflammation (P < 0.01) and inflammatory cell infiltration (P < 0.001) markedly improved by day 2 after AMT. This was associated with reduced expression (P < 0.01) and activity of MMP-8, and -9 and increased localization of TIMP-1 (P < 0.01), whereas TIMP-2 was not affected. In contrast, high levels of expression of MMP-8 and -9 remained in the cornea after tarsorrhaphy, and the TIMP-1 expression was only slightly upregulated. CONCLUSIONS: Rapid improvement of HSV-1-induced ulcerative keratitis is noted after amniotic membrane transplantation. This may be caused by reduced expression and activity of MMP-8 and -9, increased expression of TIMP-1, and sustained expression of TIMP-2. PMID- 16249484 TI - Comparison of the proliferative capacity of human corneal endothelial cells from the central and peripheral areas. AB - PURPOSE: To compare the relative proliferative capacity between human corneal endothelial cells (HCECs) cultured from the central and peripheral areas of the cornea. METHODS: Human corneas were divided into two groups based on donor age (younger group, < or =30 years of age; older group, > or =50 years of age). Corneas were trephined, and Descemet's membrane with HCECs was stripped from the central (0-6.75 mm) and peripheral (6.75-9.5 mm) areas. HCECs were then isolated from Descemet's membrane and cultivated. An equal number of passage-1 endothelial cells from each area were seeded, and the number of cells was determined at various times after seeding. Doubling times of cells from each area were compared. The antibody against minichromosome maintenance-2 (MCM2) protein was tested for replication competence. RESULTS: Morphologically, HCECs from the central area were similar to cells from the peripheral area. The doubling time (in hours) of HCECs from the central area was 35.20 in the younger group (n = 4) and 54.54 in the older group (n = 4) and from the peripheral area, 29.37 in the younger group and 46.23 in the older group. There was no significant difference (younger: P = 0.515; older: P = 0.222) between the central and peripheral area in each age group. MCM2-positive cells were consistently observed in cultures from the central, as well as peripheral, area. There was no significant difference (younger: P = 0.929; older: P = 0.613) in the percentage of MCM2-positive cells between these two areas in either age group. Even though there was no significant difference, there was a tendency toward increased doubling time and decreased percentage of MCM2 in the central area of the older group. CONCLUSIONS: These results indicate that corneal endothelial cells from both the central and peripheral areas retain potential proliferative capacity. PMID- 16249485 TI - Developmentally regulated expression of Sp1 in the mouse cornea. AB - PURPOSE: To examine the expression of transcription factor Sp1 in the cornea of the mouse eye throughout developmental stages. The environmental effect of light on Sp1 expression was also assessed. METHODS: C57BL/6 mice were set up for timed mating. Embryos on embryonic day (E)10.5, E12.5, E15.5, and E18.5 and eyes from mice on postnatal day (P)0, P7, P11, P15, P30, and P60 were collected for immunohistochemical staining and in situ hybridization. One group of mice was bred strictly in the dark between E18.5 and P15, and the eyes were collected at P0, P7, P11, and P15 time points. RESULTS: Sp1 expression was observed in the ectoderm and lens vesicle as early as E10.5. Both Sp1 protein and mRNA were abundant in the corneal basal epithelium and keratocytes until P11. Their levels were markedly reduced at P15, right after eyelid opening, and declined further between P15 and P60. In those mice bred in the dark, Sp1 was evident in the cornea at P0. The Sp1 level gradually increased until P11 and was decreased at P15. This expression pattern was nearly identical in mice bred either in a light/dark cycle or in the dark. The Sp1 level in the central lens epithelium was much lower than that in the cornea from E15.5 to late stages. CONCLUSIONS: The present study indicates that Sp1 expression is developmentally regulated, providing a basis for further investigations on the regulation of the Sp1 gene during the course of corneal development and in diseases such as keratoconus. PMID- 16249486 TI - Accelerated wound healing of alkali-burned corneas in MRL mice is associated with a reduced inflammatory signature. AB - PURPOSE: The present study was conducted to investigate healing of alkali-burned corneas in MRL/MpJ (MRL) mice. METHODS: Gross, clinical, and histologic criteria were used to compare healing of alkali-burned corneas in MRL and control C57BL/6J (B6) mice. Effects of neutrophil depletion of B6 mice and allogeneic reconstitution of B6 mice with MRL bone marrow on wound healing were evaluated. Gene expression patterns in normal and wounded corneas were surveyed with array based quantitative real-time RT-PCR (AQPCR). RESULTS: MRL mice showed accelerated reepithelialization and decreased corneal opacity compared with B6 mice after alkali wounding. Marked inflammatory cell infiltration and fibrosis were evident in the corneas and anterior chambers of B6 mice. MRL mice showed less severe lesions, except for stromal edema. Rapid reepithelialization and reduced keratitis/iritis were also observed in neutrophil-depleted B6 mice, but not in B6 mice reconstituted with MRL bone marrow. AQPCR showed transcriptional changes of fewer genes associated with inflammation and corneal tissue homeostasis in alkali burned corneas from MRL mice. Increased expression of an anti-inflammatory gene, Socs1, and a gene associated with healing, Mmp1a, were evident in MRL corneas. CONCLUSIONS: Alkali-burned corneas heal faster and more completely in MRL mice than in B6 mice, by means of rapid reepithelialization, reduced inflammation, and reduced fibrosis. Reduced inflammation, including decreased neutrophil infiltrates and the lack of a robust proinflammatory gene expression signature correlates with the rapid healing. However, the rapid-healing phenotype is not intrinsic to MRL hematopoietic progenitor cells. PMID- 16249487 TI - Color visual evoked potentials in children with type 1 diabetes: relationship to metabolic control. AB - PURPOSE: To examine the association between metabolic control (HbA(1c)) and the chromatic mechanisms of children with type 1 diabetes (T1D), by using the color visual evoked potential (VEP). METHODS: Fifty children with T1D (age range, 6 12.9 years) and 33 age-matched control subjects were tested. VEPs were recorded by placing five electrodes on the scalp according to the International 10/20 System of Electrode Placement. Active electrodes O1, O2, and Oz were placed over the visual cortex. Short-wavelength (S), and long- and medium-wavelength (LM) color stimuli consisted of vertical, photometric isoluminant (1 cyc/deg) gratings presented in a pattern onset (100 ms)-offset (400 ms) mode. Achromatic vertical gratings were presented at 3 cyc/deg. Primary outcome measure was VEP latency. The relationship between S, LM, and achromatic VEP latency, and HbA(1c) was determined by ANCOVA regression. RESULTS: S-, LM-, achromatic VEP latencies were not associated significantly with HbA(1c). Pubertal status, however, was associated significantly (P = 0.0114) and selectively with S-VEP latency. Pubertal children with T1D had delayed (mean delay, 9.5 ms) S-VEP latencies when compared with the prepubertal children with T1D. However, there was no statistically significant difference (P = 0.1573) in the effect of pubertal status on S-VEP latency between the T1D and control groups. CONCLUSIONS: Pubertal status rather than HbA(1c) appears to affect selectively the S-VEP latency of preteen children with T1D. Further study is warranted to determine whether the delay in S-VEP latency in pubertal children with T1D changes over time and whether this change could be a predictive marker for future development of background diabetic retinopathy. PMID- 16249489 TI - Development and comparison of automated classifiers for glaucoma diagnosis using Stratus optical coherence tomography. AB - PURPOSE: To develop and compare the ability of several automated classifiers to differentiate between normal and glaucomatous eyes based on the quantitative assessment of summary data reports from Stratus optical coherence tomography (OCT; Carl Zeiss Meditec Inc., Dublin, CA) in a Chinese population in Taiwan. METHODS: One randomly selected eye from each of 89 patients with glaucoma and each of 100 age- and sex-matched normal individuals were included in the study. Measurements of glaucoma variables (retinal nerve fiber layer thickness and optic nerve head analysis results) were obtained by Stratus OCT. With the Stratus OCT parameters used as input, receiver operative characteristic (ROC) curves were generated by three methods, to classify eyes as either glaucomatous or normal: linear discriminant analysis (LDA), Mahalanobis distance (MD), and artificial neural network (ANN). The area under the ROC curve was optimized by principal component analysis (PCA). Classification accuracy was determined by cross validation. RESULTS: The average visual field mean deviation was -0.7 +/- 0.6 dB in the normal group and -2.7 +/- 1.9 dB in the glaucoma group. The areas under the ROC curves were 0.824 (LDA), 0.849 (MD), 0.821 (ANN), 0.915 (LDA with PCA), 0.991 (MD with PCA), and 0.874 (ANN with PCA). CONCLUSIONS: With Stratus OCT parameters used as input, automated classifiers show promise for discriminating between glaucomatous and normal eyes. MD measured from multivariate data can predict the severity of glaucoma through the construction of a measurement space. After PCA, implementation results show that the Mahalanobis space created by MD surpasses LDA and ANN in diagnosing glaucoma. PMID- 16249490 TI - Blebbistatin, a novel inhibitor of myosin II ATPase activity, increases aqueous humor outflow facility in perfused enucleated porcine eyes. AB - PURPOSE: To investigate the specific role of myosin II, a critical biochemical determinant of cellular contraction, in modulation of aqueous humor outflow facility through the trabecular meshwork (TM) pathway. METHODS: Expression of the nonmuscle myosin II heavy chains (IIA, IIB, and IIC) in human TM and ciliary body (CB) cells was determined by RT-PCR analyses. The effects of inhibition of myosin II on cell morphology, actomyosin organization, and cell adhesions were evaluated in porcine TM and CB cells treated with blebbistatin, a cell-permeable, specific inhibitor of myosin II adenosine triphosphatase (ATPase) activity. Changes in aqueous humor outflow facility were determined in enucleated porcine eyes by using a constant-pressure Grant perfusion model system. Ultrastructural integrity of the outflow pathway in drug-perfused eyes was analyzed by transmission electron microscopy. RESULTS: Expression of nonmuscle myosin IIA and IIB was confirmed in both human TM and CB cells. Confluent cultures of primary porcine TM and CB cells treated with blebbistatin in the presence of serum revealed dose (10 200 microM)-dependent changes in cell morphology, decreases in actin stress fiber content and in focal adhesions and adherens junctions. These changes were found to be reversible within 24 hours of drug withdrawal from the cell culture media. Blebbistatin did not affect the status of myosin light chain phosphorylation in TM cells. Perfusion of enucleated porcine eyes for 5 hours with 100 and 200 microM blebbistatin produced a significant increase (P < 0.01, n = 7) in aqueous outflow facility (53% and 64%, respectively) from the baseline facility, compared with a 21% facility increase in sham control specimens. The integrity of the inner wall of aqueous plexi in drug-perfused porcine eyes was found to be intact, and TM cell morphology appeared to be similar to that noted in sham-treated eyes. CONCLUSIONS: These data demonstrate that selective inhibition of myosin II in the aqueous humor outflow pathway leads to increased aqueous outflow facility, suggesting a critical role for myosin II in the regulation of aqueous humor outflow facility. This study also suggests myosin II as a potential therapeutic target for lowering intraocular pressure in patients with glaucoma. PMID- 16249488 TI - Botulinum toxin treatment of extraocular muscles in rabbits results in increased myofiber remodeling. AB - PURPOSE: Botulinum toxin A (Botox) is commonly used for strabismus treatment. Although other muscles atrophy after intramuscular injection with Botox, extraocular muscles (EOMs) do not. A continuous process of myonuclear addition in normal uninjured adult myofibers in rabbit EOMs was studied. In this study, the effect of Botox-induced muscle paralysis on myofiber remodeling in adult EOMs was examined. METHODS: The superior rectus muscles of adult rabbits were each injected with 5 units of Botox. The contralateral muscle received injections of saline only. Bromodeoxyuridine (BrdU) was administered for various periods after Botox treatment, followed by various BrdU-free periods. Myonuclear addition, the number of BrdU-positive satellite cells, and the number of MyoD-positive satellite cells were quantified, as were alterations in expression of immature myosins. RESULTS: Intramuscular injection of Botox resulted in a significant increase in both the number of BrdU-positive myonuclei and satellite cells. MyoD expression in both satellite cells and myonuclei was significantly increased after Botox injection in EOMs. In Botox-treated EOMs, an increased number of myofibers positive for the neonatal myosin heavy chain (MyHC) isoform was detected in the orbital layer. CONCLUSIONS: Botox-induced EOM paralysis resulted in a significant short-term increase in satellite cell activation and myonuclear addition in single myofibers in adult rabbit EOMs compared with control muscles. The appearance of MyoD-positive myonuclei suggests that protein synthesis becomes upregulated after Botox injection, and this, in turn, may help explain the minimal effects on myofiber size in EOMs after Botox injection. Understanding the effect of Botox on satellite cell activation and myonuclear addition in existing myofibers may suggest new ways to maximize the clinical effectiveness of Botox in patients with strabismus. PMID- 16249491 TI - Effect of carbogen breathing and acetazolamide on optic disc PO2. AB - PURPOSE: Acetazolamide was previously shown to increase optic disc partial pressure of oxygen (PO(2)). The study was conducted to evaluate optic disc PO(2) variations during normoxia, hyperoxia (100% O(2)), and carbogen breathing (95% O(2), 5% CO(2)), before and after intravenous administration of acetazolamide. METHODS: PO(2) measurements were obtained at intervascular areas of the optic disc in nine anesthetized minipigs using oxygen-sensitive microelectrodes (10 microm tip diameter) placed at <50 microm from the optic disc. PO(2) was measured continuously during 10 minutes under normoxia, hyperoxia, or carbogen breathing. Oxygen measurements were repeated under these conditions after intravenous injection of acetazolamide (500-mg bolus). RESULTS: In hyperoxia, optic disc PO(2) increased moderately (DeltaPO(2) = 4.81 +/- 1.16 mm Hg (mean +/- SD; 24%; P < 0.001) after a much larger increase in systemic PaO(2). Carbogen breathing induced a significant increase in both systemic PaO(2) and PaCO(2), which resulted in a large increase in optic disc PO(2) (DeltaPO(2) = 13.17 +/- 2.18 mm Hg; 67%; P < 0.001). Acetazolamide induced a slow and progressive increase in both systemic PaCO(2) and optic disc PO(2) (30 minutes DeltaPO(2) = 4.24 +/- 2.45 mm Hg; 24%; P < 0.04). However, it was when carbogen was simultaneously administered that optic disc PO(2) increased most substantially (DeltaPO(2) = 18.91 +/- 5.23 mm Hg; 90%; P < 0.002). CONCLUSIONS: Carbogen breathing increases optic disc Po(2) significantly in minipigs, more than hyperoxia. The association of acetazolamide injection with carbogen breathing could induce an additional increase in optic disc PO(2) through the effect of higher systemic PaCO(2). PMID- 16249492 TI - Optical coherence tomography machine learning classifiers for glaucoma detection: a preliminary study. AB - PURPOSE: Machine-learning classifiers are trained computerized systems with the ability to detect the relationship between multiple input parameters and a diagnosis. The present study investigated whether the use of machine-learning classifiers improves optical coherence tomography (OCT) glaucoma detection. METHODS: Forty-seven patients with glaucoma (47 eyes) and 42 healthy subjects (42 eyes) were included in this cross-sectional study. Of the glaucoma patients, 27 had early disease (visual field mean deviation [MD] > or = -6 dB) and 20 had advanced glaucoma (MD < -6 dB). Machine-learning classifiers were trained to discriminate between glaucomatous and healthy eyes using parameters derived from OCT output. The classifiers were trained with all 38 parameters as well as with only 8 parameters that correlated best with the visual field MD. Five classifiers were tested: linear discriminant analysis, support vector machine, recursive partitioning and regression tree, generalized linear model, and generalized additive model. For the last two classifiers, a backward feature selection was used to find the minimal number of parameters that resulted in the best and most simple prediction. The cross-validated receiver operating characteristic (ROC) curve and accuracies were calculated. RESULTS: The largest area under the ROC curve (AROC) for glaucoma detection was achieved with the support vector machine using eight parameters (0.981). The sensitivity at 80% and 95% specificity was 97.9% and 92.5%, respectively. This classifier also performed best when judged by cross-validated accuracy (0.966). The best classification between early glaucoma and advanced glaucoma was obtained with the generalized additive model using only three parameters (AROC = 0.854). CONCLUSIONS: Automated machine classifiers of OCT data might be useful for enhancing the utility of this technology for detecting glaucomatous abnormality. PMID- 16249493 TI - Asymmetry in optic disc morphometry as measured by heidelberg retina tomography in a normal elderly population: the Bridlington Eye Assessment Project. AB - PURPOSE: To define the normal range of asymmetry in optic disc parameters measured by the Heidelberg Retina Tomograph (HRT II; Heidelberg Engineering GmbH, Dossenheim, Germany) in a normal elderly population. METHODS: Optic disc analysis of 918 eyes of 459 normal elderly patients was performed. All patients were consecutive in a cohort screened for eye disease. Normality was defined with a normal visual field on automated suprathreshold screening, intraocular pressure less than 22 mm Hg, and minimum corrected visual acuity of 6/12. Asymmetry measures were calculated by subtracting the values of the smaller disc from those of the larger disc. RESULTS: Subjects' mean age (262 female and 197 male) was 72.6 years (range, 65.5-89.3). There was no significant difference in disc area or rim area between the right and left eyes. Neither rim-to-disc area ratio asymmetry nor rim measurement asymmetries were significantly affected by age or sex. Rim-to-disc area ratio asymmetry was much less affected by the increasing difference in disc size than was absolute rim asymmetry. The 2.5th and 97.5th percentile limits of normality for the rim-to-disc area ratio asymmetry in the global and temporal-inferior analyses were -0.212 and 0.154, and -0.331 and 0.261, respectively. CONCLUSIONS: The normal range of parameter asymmetry in an age group relevant to glaucoma may be useful in the discrimination of normal from early glaucoma. Asymmetry analysis may improve discriminatory ability by reducing parameter variability based on disc size. The rim-to-disc area ratio asymmetry measure is likely to be the most useful parameter in describing normality with consistency. PMID- 16249494 TI - The effects of prostaglandin analogues on IOP in prostanoid FP-receptor-deficient mice. AB - PURPOSE: This study was designed to clarify the involvement of the prostanoid FP receptor in the intraocular pressure (IOP)-lowering effects of latanoprost, travoprost, bimatoprost, and unoprostone with the use of FP-receptor-deficient (FPKO) mice. METHODS: FPKO and wild-type (WT) mice were bred and acclimatized under a 12-hour light-dark cycle. IOP was measured under general anesthesia by a microneedle METHOD: To evaluate the effects of each drug, a single drop (3 muL) of each drug solution was topically applied in a masked manner to a randomly selected eye. IOP reduction was evaluated by the difference in IOP between the treated eye and the untreated contralateral eye in the same mouse. First, the diurnal variation and baseline IOP in WT and FPKO mice were measured. Then, to determine the window feasible for demonstrating the most marked ocular hypotensive effect, 0.005% latanoprost was applied to WT mice during the day or at night. The time when the ocular hypotensive effect was larger was selected for further studies to evaluate the effects of latanoprost (0.005%), travoprost (0.004%), bimatoprost (0.03%), and unoprostone (0.12%). In addition, bunazosin (0.1%) was also applied to demonstrate functional uveoscleral outflow in FPKO mice. All experiments were conducted under a masked study design. RESULTS: The baseline IOP (mean +/- SEM) in WT and FPKO mice was 15.0 +/- 0.2 and 15.0 +/- 0.3 mm Hg, respectively, during the day, and 18.9 +/- 0.4 and 19.2 +/- 0.4 mm Hg, respectively, at night. In WT mice, latanoprost significantly lowered IOP both during the day and at night, at 2 to 6 hours and 1 to 6 hours after application, respectively. Maximal IOP reduction was observed at 3 hours after drug instillation both during the day (10.9 +/- 1.8%) and at night (23.2 +/- 1.1%). At 3 hours after instillation, latanoprost (10.9 +/- 1.8% and 23.2 +/- 1.1%, daytime and nighttime, respectively), travoprost (15.9 +/- 1.4% and 26.1 +/- 1.2%) and bimatoprost (8.8 +/- 2.0 and 19.8 +/- 1.5%) significantly lowered IOP in WT mice both during the day and at night; isopropyl unoprostone significantly lowered IOP at night (13.7 +/- 1.9%) but not during the day (5.3 +/- 3.2%). In FPKO mice, latanoprost, travoprost, bimatoprost, and unoprostone showed no significant IOP lowering effect. Bunazosin significantly lowered IOP in both WT (22.1 +/- 1.6%) and FPKO mice (22.2 +/- 2.1%). CONCLUSIONS: A single application of latanoprost, travoprost, bimatoprost, or unoprostone had no effect on IOP in FPKO mice with presumed functional uveoscleral outflow pathways. The prostanoid FP receptor plays a crucial role in the mechanism of early IOP lowering of all commercially available prostaglandin analogues. PMID- 16249495 TI - Neuroprotection by sodium channel blockade with phenytoin in an experimental model of glaucoma. AB - PURPOSE: Sustained influx of intracellular sodium through voltage-gated sodium channels is an important event in the cascade leading to degeneration of axons. This study tested the hypothesis that sodium channel blockade with phenytoin would result in neuroprotection of retinal ganglion cells (RGCs) and optic nerve axons in an experimental model of glaucoma. METHODS: Chronic elevation of rat intraocular pressure (IOP) leading to optic nerve damage was induced using the episcleral vein occlusion model. Before induction of glaucoma, a subset of animals was placed on phenytoin-containing chow; this treatment continued for 8 weeks. Quantitative counts of backfilled RGCs and optic nerve axons was performed to examine the effects of phenytoin on glaucoma-induced adverse neurodegeneration. RESULTS: Elevated IOP resulted in a significant decrease in density of RGCs, as well as dropout of axons within the optic nerve at 8 weeks after induction. In phenytoin-treated animals, however, the loss of RGCs was significantly reduced compared to vehicle-treated glaucomatous animals. Axon loss in the optic nerve was also reduced in phenytoin-treated animals, compared to controls. CONCLUSIONS: Orally delivered phenytoin was effective in protecting neurons in an animal model of glaucoma, and merits further examination as a potential therapeutic strategy. PMID- 16249496 TI - Pathophysiologic changes in the optic nerves of eyes with primary open angle and pseudoexfoliation glaucoma. AB - PURPOSE: To determine whether differences in the optic nerve occur in eyes with primary versus secondary open-angle glaucoma. METHODS: Optic nerves obtained at autopsy from 36 eyes with primary open-angle glaucoma (POAG) and 15 with pseudoexfoliation glaucoma (PEXG) were studied quantitatively and qualitatively. Axon counts, fibrosis, capillary number and density, and arteriosclerotic changes were assessed in the postlaminar optic nerve and compared to normal age-matched autopsy eyes. Changes in composition of extracellular matrix components were evaluated by immunohistochemistry and electron microscopy. RESULTS: Marked differences were found between POAG and PEXG. Axon loss in eyes with POAG but not in PEXG was associated with increasing connective tissue in the septa and surrounding the central retinal vessels, including increased amounts of type IV and VI collagen. The total number of capillaries decreased with the loss of axons in both POAG and PEXG. POAG nerves, however, had a decrease in the density of capillaries, whereas in PEXG the capillary density did not change with axon loss. Arteriosclerotic changes were more common in glaucomatous eyes than in age matched control eyes. CONCLUSIONS: The difference in morphology of the optic nerves between POAG and PEXG indicates that in eyes with POAG, elevated IOP cannot be the only pathogenetic factor in glaucomatous optic neuropathy. Additional factors, inducing fibrosis and loss of capillaries, seem to be involved. Such additional factors may also contribute to the clinical finding in POAG that nerves can become damaged without elevation of intraocular pressure. PMID- 16249497 TI - Relationships between standard automated perimetry, HRT confocal scanning laser ophthalmoscopy, and GDx VCC scanning laser polarimetry. AB - PURPOSE: This study was designed to determine and compare the relationships between visual function measured with standard automated perimetry (SAP) and structure, either as neuroretinal rim area measured with confocal scanning laser ophthalmoscopy (CSLO), or as retinal nerve fiber layer thickness determined by scanning laser polarimetry with variable corneal compensation (SLP-VCC). METHODS: Forty-six healthy subjects and 76 glaucoma patients were examined with SAP, with CSLO by means of the commercially available Heidelberg Retina Tomograph I (HRT), and with SLP-VCC by means of the commercially available GDx VCC. The relationships between SAP, expressed either in the typically used decibel scale or as number of abnormal points in the total deviation probability plot, and CSLO and between SAP and SLP-VCC were described with linear and logarithmic regression analysis for global data and six individual sectors. The relationship between measurements with CSLO and SLP-VCC was fit with linear regression analysis. RESULTS: The relationships between SAP and CSLO and between SAP and SLP-VCC appeared curvilinear for all sectors except the temporal one between SAP and SLP VCC. For CSLO, a logarithmic fit was significantly better than a linear one for the global data and in the superotemporal and inferonasal sectors. For SLP-VCC, a curvilinear fit was better for the global data and in the superotemporal, superonasal, and inferonasal sectors. CSLO data correlated linearly with SLP-VCC data in all sectors, except temporally. CONCLUSIONS: CSLO and SLP-VCC showed a very similar curvilinear relationship with SAP. The observed curvilinear relationships confirm earlier reports that these imaging devices appear to detect glaucomatous loss earlier than SAP. PMID- 16249498 TI - Factors influencing optic nerve head biomechanics. AB - PURPOSE: The biomechanical environment within the optic nerve head (ONH) may play a role in retinal ganglion cell loss in glaucomatous optic neuropathy. This was a systematic analysis in which finite element methods were used to determine which anatomic and biomechanical factors most influenced the biomechanical response of the ONH to acute changes in IOP. METHODS: Based on a previously described computational model of the eye, each of 21 input factors, representing the biomechanical properties of relevant ocular tissues, the IOP, and 14 geometric factors were independently varied. The biomechanical response of the ONH tissues was quantified through a set of 29 outcome measures, including peak and mean stress and strain within each tissue, and measures of geometric changes in ONH tissues. Input factors were ranked according to their aggregated influence on groups of outcome measures. RESULTS: The five input factors that had the largest influence across all outcome measures were, in ranked order: stiffness of the sclera, radius of the eye, stiffness of the lamina cribrosa, IOP, and thickness of the scleral shell. The five least influential factors were, in reverse ranked order: retinal thickness, peripapillary rim height, cup depth, cup-to-disc ratio, and pial thickness. Factor ranks were similar for various outcome measure groups and factor ranges. CONCLUSIONS: The model predicts that ONH biomechanics are strongly dependent on scleral biomechanical properties. Acute deformations of ONH tissues, and the consequent high levels of neural tissue strain, were less strongly dependent on the action of IOP directly on the internal surface of the ONH than on the indirect effects of IOP on the sclera. This suggests that interindividual variations in scleral properties could be a risk factor for the development of glaucoma. Eye size and lamina cribrosa biomechanical properties also have a strong influence on ONH biomechanics. PMID- 16249500 TI - Silencing Toll-like receptor-9 in Pseudomonas aeruginosa keratitis. AB - PURPOSE: To determine the effects of silencing Toll-like receptor (TLR) 9 signaling in Pseudomonas aeruginosa keratitis. METHODS: Corneal TLR9 mRNA levels were tested by RT-PCR in C57BL/6 (B6, susceptible) and BALB/c (resistant) mice and compared. The response of B6 mice to CpG DNA, which binds TLR9, was tested after subconjunctival injection of mice with control or CpG DNA; TLR9, IL-1beta, macrophage inflammatory protein (MIP)-2, IL-4, IL-10, IL-12, IL-18, and IFN-gamma levels were measured by RT-PCR. Langerhans cells (LCs) were stimulated with CpG DNA and treated with TLR9 or control siRNA, and mRNA levels of TLR9, IL-1beta, and MIP-2 were detected by RT-PCR. In addition, IL-1beta levels were tested by ELISA. Then B6 mice were injected subconjunctivally with control or TLR9 siRNA before infection and treated topically afterward. Slit lamp, clinical score, RT PCR, ELISA, myeloperoxidase assay, and plate counts were performed. RESULTS: TLR9 mRNA levels were sixfold higher in B6 than in BALB/c corneas the day after injection. B6 mice injected with CpG DNA exhibited an increase in corneal mRNA for TLR9, IL-1beta, MIP-2, IL-12, and IFN-gamma over controls. LCs stimulated with CpG DNA and treated with TLR9 siRNA exhibited reduced TLR9, IL-1beta, and MIP-2 levels compared with controls. Finally, B6 mice treated with TLR9 siRNA showed decreases in corneal opacity, polymorphonuclear leukocyte number, IL-12 and IFN-gamma mRNA, IL-1beta, and MIP-2 protein compared with those treated with control siRNA. Fewer corneas perforated in these mice, but bacterial loads were higher than in controls. CONCLUSIONS: Signaling through TLR9 appears important in P. aeruginosa keratitis, and silencing TLR9 signaling reduces inflammation but likely contributes to decreased bacterial killing in the cornea. PMID- 16249499 TI - Fate maps of neural crest and mesoderm in the mammalian eye. AB - PURPOSE: Structures derived from periocular mesenchyme arise by complex interactions between neural crest and mesoderm. Defects in development or function of structures derived from periocular mesenchyme result in debilitating vision loss, including glaucoma. The determination of long-term fates for neural crest and mesoderm in mammals has been inhibited by the lack of suitable marking systems. In the present study, the first long-term fate maps are presented for neural crest and mesoderm in a mammalian eye. METHODS: Complementary binary genetic approaches were used to mark indelibly the neural crest and mesoderm in the developing eye. Component one is a transgene expressing Cre recombinase under the control of an appropriate tissue-specific promoter. The second component is the conditional Cre reporter R26R, which is activated by the Cre recombinase expressed from the transgene. Lineage-marked cells were counterstained for expression of key transcription factors. RESULTS: The results established that fates of neural crest and mesoderm in mice were similar to but not identical with those in birds. They also showed that five early transcription factor genes are expressed in unique patterns in fate-marked neural crest and mesoderm during early ocular development. CONCLUSIONS: The data provide essential new information toward understanding the complex interactions required for normal development and function of the mammalian eye. The results also underscore the importance of confirming neural crest and mesoderm fates in a model mammalian system. The complementary systems used in this study should be useful for studying the respective cell fates in other organ systems. PMID- 16249501 TI - M cells in the follicle-associated epithelium of the rabbit conjunctiva preferentially bind and translocate latex beads. AB - PURPOSE: This study investigates the presence of M cells in the rabbit conjunctiva. Resolving whether the conjunctiva contains M cells is important, because at other mucosal sites, these antigen sampling cells are known to initiate the mucosal immune response and to act as a site of entry for opportunistic pathogens. METHODS: Fluorescent 0.2-microm polystyrene latex beads were either instilled into the conjunctival sac for 20 to 120 minutes in vivo or applied to flat mounts in vitro. Specimens were assessed by epi-fluorescence stereomicroscopy, widefield fluorescence microscopy, confocal scanning laser microscopy, and transmission and scanning electron microscopy. RESULTS: Latex beads preferentially bound to a subpopulation of cells in the epithelium overlying mucosal lymphoid follicles in the fornix region. At 4 degrees C, the beads were associated with the apical surface of cells that had longer, more irregular microvilli than the surrounding epithelial cells. Within 20 minutes of an in vivo exposure, latex beads were internalized by the follicle-associated epithelial cells and with time moved into the underlying follicle region. After 120 minutes of in vivo exposure, latex beads could be found in cervical lymph nodes. CONCLUSIONS: This study demonstrates that the follicle-associated epithelium of the rabbit conjunctiva contains a cell with morphologic characteristics and the ability to bind and translocate latex beads, which make it indistinguishable from antigen sampling M cells in the rabbit cecum and tonsils. Consistent with its hypothesized antigen sampling role, beads that have been translocated by this cell are rapidly transferred to cervical lymph nodes. PMID- 16249502 TI - Microarray analysis of cytokine and chemokine gene expression after prednisolone treatment in murine experimental autoimmune uveoretinitis. AB - PURPOSE: The purpose of this study was to investigate changes in gene expression of cytokines and chemokines and their receptors after systemic prednisolone treatment in experimental autoimmune uveoretinitis (EAU). METHODS: EAU mice received one intravenous injection of 7.5 mg/kg prednisolone (PDS) sodium phosphate at the peak of inflammation. EAU mice treated with only solvent served as the control. Total RNA was extracted from the whole eyes 1, 2, and 3 days after treatment. Gene expression analysis was conducted with a cDNA microarray, which contains 117 individual transcripts encoding the genes of the cytokines and chemokines and their receptors (29 cytokines and 34 cytokine receptors; 33 chemokines and 21 chemokine receptors). Comparisons of expression between PDS treated and placebo-treated EAU mice at each time point were performed. The genes were sorted into clusters based on expression profiles, to clarify the gene regulation pattern after treatment. RESULTS: Forty-seven genes had a significant decrease in expression 1 day after treatment, 10 genes on day 2, and 46 genes on day 3. Ten genes were upregulated on day 1, but no gene was upregulated thereafter. Hierarchical cluster analysis of the microarray demonstrated that gene expression changes in EAU after treatment with PDS showed four patterns: flat, mountain, steep downhill, and less steep downhill. CONCLUSIONS: The implications of the clusters after treatment with PDS remain unclear. The results showed that hierarchical cluster analysis based on comprehensive gene expression profiles may provide a powerful tool for identifying genes not previously associated with the therapeutic targets in ocular inflammation. PMID- 16249503 TI - LBP and CD14 secreted in tears by the lacrimal glands modulate the LPS response of corneal epithelial cells. AB - PURPOSE: Lipopolysaccharide (LPS) is one of the most powerful bacterial virulence factors in terms of proinflammatory properties and is likely to contribute to corneal bacterial keratitis. Better understanding of the spatial expression of the LPS receptor components at the tear-corneal interface might facilitate enhanced functions of the LPS receptor complex in ocular defense against Gram negative infections. METHODS: The expression of LPS-binding protein (LBP), CD14, toll-like receptor (TLR)-4, and MD-2 in human lacrimal glands, reflex tears, and corneal epithelia was examined by ELISA, RT-PCR, Western blot analysis, and immunofluorescence. The release of proinflammatory cytokines after the activation of primary and immortalized corneal epithelial cells with LPS and human tears was measured by ELISA. RESULTS: LBP and CD14 proteins were detected in reflex human tears. Human lacrimal glands and corneal epithelia expressed LBP, CD14, TLR4, and MD-2 mRNAs and proteins. In the corneal epithelium, LBP was mainly expressed by superficial and basal epithelial cells, whereas CD14, TLR4, and MD-2 expression were limited to the wing and basal epithelial cells. In a dose-dependant manner, tear CD14 and LBP mediated the secretion of interleukin (IL)-6 and IL-8 by corneal epithelia cells when challenged with LPS. CONCLUSIONS: Tear CD14 and LBP complemented the LPS receptor complex expressed by the corneal epithelia to trigger an immune response in the presence of LPS. The complementation of these tear and corneal immune proteins could play an important role in LPS recognition and signaling and, therefore, could modulate ocular innate immunity. PMID- 16249504 TI - Cytokine gene polymorphism in sympathetic ophthalmia. AB - PURPOSE: Sympathetic ophthalmia (SO) is a prototypical autoimmune disease in which injury to one eye causes sight-threatening inflammation in the otherwise normal contralateral eye. Previous work found that human leukocyte antigen alleles HLA-DRB1*04 and DQA1*03 are markers of increased susceptibility and severity in British and Irish patients. Evidence is accumulating that single nucleotide polymorphisms (SNPs) in cytokine genes can also influence the development of autoimmune disease through their effect on levels of cytokine production. The purpose of this study was to determine whether polymorphisms in the cytokine genes are important markers for disease severity and outcome in patients with SO. METHODS: Twenty-six British and Irish patients meeting well defined criteria for the diagnosis of SO were compared with 48 matched controls. Genotyping of SNPs in the TNFalpha, TNFbeta, and IL-10 genes was performed using the polymerase chain reaction and sequence-specific primers (SSP-PCRs) and of the CTLA-4 and TNF receptor 2 genes using restriction length polymorphism-PCR (RFLP PCR). RESULTS: Significant associations were found between the IL-10 -1082 SNP and disease recurrence from previously stable disease and the level of steroids required for maintenance therapy. In addition, the GCC IL-10 promoter haplotype (IL-10 -1082G, -819C, -592C) was found to be protective against disease recurrence. CONCLUSIONS: These results show that cytokine gene polymorphisms are markers for the severity of disease in SO. They were found to be associated with recurrence of previously stable disease and with the level of maintenance steroid treatment required to control inflammatory activity. PMID- 16249505 TI - Multiplex bead immunoassay analysis of aqueous humor reveals distinct cytokine profiles in uveitis. AB - PURPOSE: To extensively characterize the complex network of cytokines present in uveitis aqueous humor (AqH), and the relationships between cytokines and the cellular infiltrate. METHODS: AqH from noninflammatory control subjects and patients with idiopathic, Fuchs' heterochromic cyclitis (FHC), and herpes-viral or Behcet's uveitis were analyzed for IL-1beta, -2, -4, -5, -7, -8, -10, -12, 13, -15, TNFalpha, IFNgamma, CCL2 (MCP-1), CCL5 (RANTES), CCL11 (Eotaxin), TGFbeta2, and CXCL12 (SDF-1), using multiplex bead immunoassays. The cellular infiltrate was also determined for each sample. RESULTS: Idiopathic uveitis AqH, compared with noninflammatory controls, was characterized by high levels of IL-6, IL-8, CCL2 and IFNgamma, the levels of which correlated with each other. For IL-6 and IL-8 these levels were proportional to the number of neutrophils present. By contrast, the levels of both TGFbeta2 and CXCL12 decreased in idiopathic uveitis AqH with increasing inflammation. Cluster analysis showed a degree of segregation between noninflammatory and idiopathic uveitis AqH. Further examination using random forest analysis yielded a complete distinction between these two groups. The minimum cytokines required for this classification were IL-6, IL-8, CCL2, IL 13, TNFalpha, and IL-2. CONCLUSIONS: Application of multiplex bead immunoassays has allowed us to identify distinct patterns of cytokines that relate to both clinical disease and the cellular infiltrates present. Bioinformatics analysis allowed identification of cytokines that differentiate idiopathic uveitis from noninflammatory control AqH and are likely to be important for the pathogenesis of uveitis. PMID- 16249506 TI - Effect of TGF-beta2 and anti-TGF-beta2 antibody in a new in vivo rodent model of posterior capsule opacification. AB - PURPOSE: This study evaluated the effect of transforming growth factor (TGF) beta2 and anti-TGF-beta2 antibody in a rodent model of posterior capsule opacification (PCO). METHODS: An extracapsular lens extraction (ECLE) was performed in 72 Sprague-Dawley rats. At the end of the procedure, 10 microL TGF beta2 (TGF-beta2-treated group), fetal calf serum (FCS)/phosphate-buffered saline (PBS; FCS/PBS-treated control group), a human monoclonal TGF-beta2 antibody (anti TGF-beta2-treated group), or a null control IgG4 antibody (null antibody-treated control group) was injected into the capsule. Animals were killed 3 and 14 days postoperatively. Eyes were evaluated clinically prior to euthanatization, then enucleated and processed for light microscopy and immunohistochemistry afterward. PCO was evaluated clinically and histopathologically. Student's t-test and chi(2) were used to assess differences between groups. RESULTS: There were no statistically significant clinical or histopathological differences in degree of PCO between the TGF-beta2- and FCS/PBS-treated groups at 3 and 14 days after ECLE. Nor were there differences between the anti-TGF-beta2- and the null antibody-treated groups, with the exception of the histopathology score for capsule wrinkling 3 days after ECLE (P = 0.02). alpha-Smooth-muscle actin staining was observed in the lens capsular bag only in areas where there was close contact with the iris. CONCLUSIONS: No sustained effect of TGF-beta2 or anti-TGF-beta2 antibody on PCO was found in rodents at the dose and timing administered in this study. Iris cells may play a role in the process of epithelial mesenchymal transition linked to PCO. PMID- 16249507 TI - Parasympathetic mediated pupillary dilation elicited by lingual nerve stimulation in cats. AB - PURPOSE: To determine the autonomic efferent nerve pathways for the reflex pupillary dilation elicited by somatic stimulation in cats. METHODS: Cats anesthetized with a mixture of alpha-chloralose (50 mg/kg) and urethane (100 mg/kg) were intubated and paralyzed by intravenous injection of pancuronium bromide. The central cut end of the lingual nerve (LN) was stimulated electrically to simulate somatic stimulation, and 1 microL of lidocaine (2%) was microinjected into the Vsp or the EW nucleus to determine its effect on the pupillary dilation induced by LN stimulation. The effect of electrically stimulating the Vsp or sectioning the superior cervical sympathetic nerve (CSN) on the pupillary response was also examined. RESULTS: Stimulation of the LN or the trigeminal spinal nucleus (Vsp) evoked pupillary dilation in a frequency- and intensity-dependent manner. These responses were not affected by sectioning the ipsilateral or both CSNs. The pupillary responses were markedly suppressed by microinjecting lidocaine into the ipsilateral Vsp or the Edinger-Westphal (EW) nucleus, but not by injection into the contralateral Vsp. CONCLUSIONS: These results indicate that the Vsp and EW nucleus act as bulbar relay centers for pupillary dilation elicited by LN stimulation and suggest that the efferent arc of the response is a parasympathetic pathway. The contralateral pupillary dilation appears to be mediated, at least in part, by fibers projecting from the Vsp to the contralateral EW nucleus. PMID- 16249508 TI - Inhaled carbon monoxide increases retinal and choroidal blood flow in healthy humans. AB - PURPOSE: It has been hypothesized that carbon monoxide (CO) acts as an important vascular paracrine factor and plays a role in blood flow regulation in several tissues. The present study investigated the effect of inhaled CO on retinal and choroidal blood flow. METHODS: Fifteen healthy male volunteers were studied in a randomized, double-masked, placebo-controlled design with washout periods of at least 1 week between study days. CO in a dose of 500 ppm or placebo (synthetic air without CO) was inhaled for 60 minutes. Ocular hemodynamics were measured at baseline and at 30 and 60 minutes after start of inhalation. Retinal vessel diameters were measured with a retinal vessel analyzer. RBC velocity was assessed using bidirectional laser Doppler velocimetry. Retinal blood flow was calculated based on retinal vessel diameters and RBC velocity. Fundus pulsation amplitude (FPA) was measured using laser interferometry, and submacular choroidal blood flow using laser Doppler flowmetry. RESULTS: Breathing of CO significantly increased carboxyhemoglobine, from 1.2 +/- 0.5% to 8.5 +/- 0.9% and 9.4 +/- 0.6% at the two time points, respectively (P < 0.01). The diameter of retinal arteries increased by +3.5 +/- 3.8% and +4.2 +/- 3.9% (P < 0.01) in response to CO inhalation. In retinal veins, CO also induced an increase in diameter of +4.3 +/- 3.0% and +4.8 +/- 5.0%, respectively (P < 0.01). By contrast, placebo did not influence retinal vessel diameter. RBC velocity tended to increase during CO inhalation (+8 +/- 22%), but this effect did not reach the level of significance (P = 0.1). Calculated retinal blood flow increased significantly by +12 +/- 5% (P < 0.02). FPA increased after breathing CO by +20 +/- 20% and +26 +/- 21% at the two time points, respectively (P < 0.01). Subfoveal choroidal blood flow increased by +14 +/- 9% and +15 +/- 9% during breathing of CO (P < 0.01). CONCLUSIONS: This experiment demonstrated that retinal and choroidal blood flow increase during inhalation of CO. Whether this increase is caused by tissue hypoxia or a yet unknown mechanism has to be clarified. PMID- 16249509 TI - Vascular damage in a mouse model of diabetic retinopathy: relation to neuronal and glial changes. AB - PURPOSE: Lack of information about the development of diabetic retinopathy in mice has greatly hindered the use of genetic mouse models for the study of disease mechanisms and the development of therapeutic strategies. The objective of this study was to characterize the occurrence and pathologic progression of diabetic retinopathy in C57Bl/6J mice. METHODS: Diabetes was induced with five consecutive injections of streptozotocin (STZ). The retinas were collected at different time points (2 weeks to 22 months) after the induction of diabetes and examined by using molecular, histologic, and immunohistochemical techniques and morphometric analysis. RESULTS: There was transient induction of cell apoptosis and caspase-3 activation in retinal neurons of C57Bl/6 mice within days of diabetes induction. Glial fibrillary acidic protein (GFAP), a marker of glial activation, likewise was transiently upregulated, seemingly in astrocytes but not in Muller cells. These abnormalities quickly returned to normal; ultimately, no detectable loss of retinal ganglion cells (RGCs) was noted by any of three independent methods (number of cells in ganglion cell layer of retinal cross sections, retrograde labeling of retinal ganglion cells with fluorescent dye, or TUNEL staining) after up to a 1-year duration of diabetes. Despite this apparent lack of evidence for progressive damage in neurons and glial cells, diabetic mice developed vascular disease characteristic of the early stage of diabetic retinopathy beginning at 6 months after the onset of disease. The vascular damage formation of acellular capillaries and pericyte ghosts-continued to increase through the 18 months examined. CONCLUSIONS: Diabetic C57Bl/6J mice develop capillary lesion that are characteristic of the early stages of diabetic retinopathy in patients. The data suggest that diabetes-induced degeneration of retinal capillaries can develop independent of neuronal loss or chronic GFAP upregulation in glial cells. PMID- 16249510 TI - Expression of the Opa1 mitochondrial protein in retinal ganglion cells: its downregulation causes aggregation of the mitochondrial network. AB - PURPOSE: Mutations in the mitochondrial dynamin-related GTPase OPA1 cause autosomal dominant optic atrophy (ADOA), but the pathophysiology of this disease is unknown. As a first step in functional studies, this study was conducted to evaluate the expression of Opa1 in whole retina and in isolated retinal ganglion cells (RGCs) and to test the effects of Opa1 downregulation in cultured RGCs. METHODS: Opa1 mRNA isoforms from total retina and from RGCs freshly isolated by immunopanning were determined by RT-PCR. Protein expression was examined by immunohistochemistry and Western blot with antibodies against Opa1 and cytochrome c, and the mitochondrial network was visualized with a mitochondrial marker. Short interfering (si)RNA targeting OPA1 mRNAs were transfected to cultured RGCs and mitochondrial network phenotypes were followed for 15 days, in comparison with those of cerebellar granule cells (CGCs). RESULTS: Opa1 expression did not predominate in rat postnatal RGCs as found by immunohistochemistry and Western blot analysis. The pattern of mRNA isoforms was similar in whole retina and RGCs. After a few days in culture, isolated RGCs showed fine mitochondrial punctiform structures in the soma and neurites that colocalized with cytochrome c and Opa1. Opa1 knockdown in RGCs induced mitochondrial network aggregation at a higher rate than in CGCs. CONCLUSIONS: Results suggest that the level of expression and the mRNA isoforms do not underlie the vulnerability of RGCs to OPA1 mutations. However, aggregation of the mitochondrial network induced by the downregulation of Opa1 appears more frequent in RGCs than in control CGCs. PMID- 16249511 TI - Serum inflammatory markers in diabetic retinopathy. AB - PURPOSE: To evaluate the association of serum factors with the severity of diabetic retinopathy and to assess their presence in retinal tissue obtained at autopsy. METHODS: The following serum factors of 93 subjects were examined at the National Eye Institute (NEI) clinical center: the chemokines regulated on activation, normal T-cell expressed and presumably secreted (RANTES)/CCL5, epithelial neutrophil activator (ENA)-78/CXCL5, interferon-induced protein (IP) 10/CXCL10, stromal cell-derived factor (SDF)-1alpha/CXCLl2, monocyte chemoattractant protein (MCP)-1/CCL2, macrophage inflammatory protein (MIP) 1alpha/CCL3, interleukin (IL)-8/CXCL8; the cytokine IL-6; the cell adhesion molecules intercellular adhesion molecule (ICAM-1/CD54) and vascular cell adhesion molecule (VCAM/CD106); and the growth factor vascular endothelial growth factor (VEGF). Logistic regression was performed to assess the association of these factors with age, sex, severity of retinopathy, hemoglobin A(1C), total cholesterol, creatinine, duration of diabetes, and presence of macular edema. The outcome assessed was severity of retinopathy. Frozen sections of two donor eyes obtained at autopsy from a donor with documented severe nonproliferative diabetic retinopathy and diabetic macular edema and of a normal nondiabetic eye were processed by immunoperoxidase staining with primary antibodies against RANTES, MCP-1, ICAM-1, and LFA-1alpha/CD11a. RESULTS: The levels of RANTES and SDF-1alpha were significantly elevated in patients with at least severe nonproliferative diabetic retinopathy compared with those with less severe diabetic retinopathy (P < 0.001 and 0.007, respectively). Positive immunostaining was observed in the inner retina for MCP-1 and RANTES of the patient with diabetes. Staining was strongly positive throughout the diabetic retina for ICAM-1. Normal retinal tissues showed little reactivity. CONCLUSIONS: Serum chemokines were significantly elevated in patients with at least severe nonproliferative diabetic retinopathy compared with those who had less severe retinopathy. Elevated levels of the chemokines and cell adhesion molecules were also identified in eyes of a donor with ischemic diabetic retinopathy. These findings provide evidence to support the role of inflammation in the pathogenesis of diabetic retinopathy. PMID- 16249513 TI - Hepatocyte growth factor protects RPE cells from apoptosis induced by glutathione depletion. AB - PURPOSE: To study the mechanism of the protective effect of hepatocyte growth factor (HGF) in oxidative injury to RPE cells induced by glutathione (GSH) depletion. METHODS: RPE cells were treated with HGF for 24 hours (20 ng/mL) and then were treated with DL-buthionine-(S,R)-sulfoximine (BSO) for an additional 24 hours. Cell death, apoptosis, and GSH levels were measured. Levels of intracellular reactive oxygen species (ROS) and their cellular localization were assessed by confocal microscopy. Expression of Bcl-2 and release of cytochrome c from mitochondria were quantified. The effect of BSO on caspase-3 activation and expression was determined. Gene expression of key enzymes of GSH metabolism by real-time PCR and regulation and translocation of the transcription factor NF-E2 related factor (Nrf2) by BSO were examined. RESULTS: Treatment with BSO-induced apoptosis in RPE caused a significant decrease in intracellular GSH and in GSH/GSSG ratios. Marked increases in lipid peroxidase (LPO), ROS, and mitochondrial cytochrome c release and a decrease in Bcl-2 expression were observed. Elevated GSH/GSSG ratio (especially in mitochondria), decreased LPO and ROS, attenuation of apoptosis, and partial restoration of Bcl-2 expression were found in the HGF-pretreated cells. BSO activated caspase-3, and this effect was significantly blocked by HGF. Both HGF and BSO induced anti-oxidant gene expression. Nrf2 translocated to the nuclear region after treatment with BSO, whereas HGF did not induce such translocation. CONCLUSIONS: The protective effect of HGF may be attributed in part to the elevation of mitochondrial GSH. BSO and HGF act in concert to enhance GSH-related gene expression in stressed RPE cells. PMID- 16249512 TI - (R)-alpha-lipoic acid protects retinal pigment epithelial cells from oxidative damage. AB - PURPOSE: To determine whether (R)-alpha-lipoic acid (LA) protects cultured human fetal retinal pigment epithelial (hfRPE) cells against oxidative injury and identify the pathways that may mediate protection. METHODS: Cultured hfRPE cells were pretreated with various concentrations of LA for 14 to 16 hours followed by treatment with a chemical oxidant, tert-butylhydroperoxide (t-BuOOH; 0.8 mM, 3 hours). Reactive oxygen species (ROS) production and cell viability were measured using H(2)DCF and MTT assays, respectively. RPE cells were evaluated with fluorescent dyes (SYTOX Orange and SYTO Green; Molecular Probes, Eugene, OR), which differentiate between live and dead cells. Apoptosis was visualized by using the TUNEL assay. Changes in mitochondrial membrane potential were detected by JC-1 dye. Intracellular levels of reduced glutathione (GSH) and oxidized glutathione (GSSG) were measured by HPLC. Regulation of gamma-glutamylcysteine ligase (GCL), the rate-controlling enzyme of GSH production, was assayed by RT PCR. RESULTS: Pretreatment of hfRPE cells with LA, 0.2 mM and 0.5 mM, significantly reduced the levels of t-BuOOH-induced intracellular ROS, by 23% and 49%, respectively. LA (0.5 mM) prevented oxidant-induced cell death and apoptosis and also increased the viability of oxidant-treated hfRPE cells from 38% to 90% of control. LA upregulated the mRNA expression of GCL, and was protective against t-BuOOH-induced decreases in both mitochondrial membrane potential and intracellular levels of GSH and GSH/GSSG. CONCLUSIONS: The present study suggests that the protective effect of LA involves multiple pathways and that LA could be effective against age-associated increase in oxidative stress and mitochondrial dysfunction in RPE cells. PMID- 16249514 TI - A critical role of CaBP4 in the cone synapse. AB - PURPOSE: CaBP4, a photoreceptor-specific protein of the rods and cones, is essential for the development and maintenance of the mouse photoreceptor synapse. In this study, double CaBP4/rod alpha-transducin knockout (Cabp4(-/-)Gnat1(-/-)) mice lacking the rod-mediated component of electrophysiologic responses were generated and analyzed to investigate the role of CaBP4 in cones. METHODS: The retinal morphology and physiologic function of 2-month-old Cabp4(-/-)Gnat1(-/-) mice were analyzed using immunocytochemistry, electron microscopy, and single flash and flicker electroretinography (ERG). RESULTS: The thickness of the outer plexiform layer and the number of photoreceptor terminals in Cabp4(-/-)Gnat1(-/-) mice were reduced to levels similar to those of Cabp4(-/-) mice. Single-flash and flicker ERG showed that the amplitude and sensitivity of the b-wave in the Cabp4( /-)Gnat1(-/-) mice were severely attenuated compared with those in wild-type and Gnat1(-/-) mice. CONCLUSIONS: Results indicate that the cone synaptic function in Cabp4(-/-)Gnat1(-/-) mice was severely disrupted, whereas the morphologic defects observed in Cabp4(-/-)Gnat1(-/-) mice were similar to those of single Cabp4(-/-) knockout mice. This and a previous study reveal that CaBP4 is critical for signal transmission from rods and cones to second-order neurons. PMID- 16249515 TI - Mutations in GRM6 cause autosomal recessive congenital stationary night blindness with a distinctive scotopic 15-Hz flicker electroretinogram. AB - PURPOSE: Congenital stationary night blindness (CSNB) is a group of nonprogressive retinal disorders characterized by impaired night vision that occurs in autosomal dominant, autosomal recessive, or X-linked forms. Autosomal recessive (ar)CSNB seems to be very rare. Mice lacking the metabotropic glutamate receptor 6 (Grm6) have a defect in signal transmission from the photoreceptors to ON-bipolar cells. In the current study, the human orthologue (GRM6) was screened as a likely candidate for arCSNB. METHODS: arCSNB individuals of five families were screened for mutations in GRM6. Subsequently, they were examined with standard and 15-Hz flicker electroretinography (ERG). These recordings were compared with those of patients with X-linked CSNB1. RESULTS: Affected individuals in three of five families carried either compound heterozygous or homozygous mutations in GRM6. Strikingly, all of them displayed a distinctive abnormality of the rod pathway signals on scotopic 15-Hz flicker ERG. CONCLUSIONS: The novel profile identified in this study suggests the existence of more than two rod pathways. The distinctive ERG feature was not observed in patients with X-linked CSNB1 and additional affected individuals with unknown molecular defect. These observations will help to discriminate autosomal recessive from X-linked recessive cases by ERG and molecular genetic analysis. PMID- 16249516 TI - Triamcinolone acetonide destabilizes VEGF mRNA in Muller cells under continuous cobalt stimulation. AB - PURPOSE: To identify the molecular mechanism of steroid-induced downregulation of vascular endothelial growth factor (VEGF) synthesis in Muller cells. METHODS: Confluent cultures of human Muller cells (MIO-M1) were treated with 100 microM CoCl(2), 1 microg/mL triamcinolone acetonide (TA), or both. VEGF secretion was measured with respect to time by ELISA. VEGF mRNA quantity and stability were analyzed by reverse transcriptase-polymerase chain reaction. The activity of hypoxia-inducible factor (HIF)-1 was measured by the relative binding of HIF-1 protein to the hypoxia response element (HRE), by gel shift and ELISA. The HIF 1alpha protein level was determined with Western blot. RESULTS: TA decreased VEGF secretion by at least 50% in the presence of continuous cobalt stimulus. VEGF mRNA decreased 50- to 100-fold 6 hours after treatment with TA and cobalt compared with cobalt alone. VEGF mRNA stability was decreased in cobalt stimulated, TA-treated cells compared with cobalt alone in cells synchronized by exposure to actinomycin D. HIF-1alpha protein level was sustained for the entire 24-hour treatment period and partitioned into nuclear, not cytosolic, fractions. HIF-1 activity was decreased by 20% to 30% in the presence of TA and cobalt compared with cobalt alone. CONCLUSIONS: TA may decrease VEGF synthesis by nongenomic destabilization of VEGF mRNA in cobalt-stimulated Muller cells. There was little effect on the total HIF-1alpha protein level, HIF-1 partitioning, and HIF-1 activity. PMID- 16249518 TI - When red lights look yellow. AB - PURPOSE: Red signals are typically used to signify danger. This study was conducted to investigate a situation identified by train drivers in which red signals appear yellow when viewed at long distances (approximately 900 m) through progressive-addition lenses. METHODS: A laboratory study was conducted to investigate the effects of defocus, target size, ambient illumination, and surround characteristics on the extent of the color misperception of train signals by nine visually normal participants. The data from the laboratory study were validated in a field study by measuring the amounts of defocus and the distances at which the misperception of the color of train signals was apparent and whether these distances varied as a function of time of day. RESULTS: The laboratory study demonstrated that small red targets (approximately 1 min arc) can appear yellow when viewed through small amounts of defocus (approximately +0.75 D) under bright illumination (1910 cd/m(2)). In the field study, the defocus needed to produce the color misperception was similar to that found in the laboratory study. Time of day affected the color misperception, and there was no misperception at night. CONCLUSIONS: The color misperception is not solely associated with progressive-addition lenses, but occurs in the presence of small amounts of positive defocus. The potential for the misperception to result in collisions and fatalities presents a major safety concern. PMID- 16249520 TI - Framework for environmental exposure research: the disease-first approach. PMID- 16249517 TI - Anti-inflammatory effect of docosahexaenoic acid on cytokine-induced adhesion molecule expression in human retinal vascular endothelial cells. AB - PURPOSE: Docosahexaenoic acid (DHA(22:6n3)), the principal n3-polyunsaturated fatty acid (PUFA) in the retina, has been shown to have a pronounced anti inflammatory effect in numerous in vivo and in vitro studies. Despite the importance of vascular inflammation in diabetic retinopathy, the anti inflammatory role of DHA(22:6n3) in cytokine-stimulated human retinal vascular endothelial cells (hRVECs) has not been addressed. METHODS: Cytokine-induced expression of cell adhesion molecules (CAMs) was assessed by Western blot. The effect of DHA(22:6n3) on cytokine-induced nuclear factor (NF)-kappaB signaling was analyzed by Western blot analysis and electrophoretic mobility shift assay (EMSA). RESULTS: Stimulation of hRVECs with VEGF(165), TNFalpha, or IL-1beta for 6 to 24 hours caused significant induction of intracellular adhesion molecule (ICAM)-1 and vascular cell adhesion molecule (VCAM)-1 expression. Pretreatment of the cells with 100 microM of BSA-bound DHA(22:6n3) for 24 hours remarkably inhibited cytokine-induced CAM expression. IL-1beta, TNFalpha, and VEGF(165) induced nuclear translocation and binding of p65 and p50 NF-kappaB isoforms to the VCAM-1 promoter. DHA(22:6n3) pretreatment inhibited cytokine-induced NF kappaB binding by 25% to 40%. Moreover, DHA(22:6n3) diminished IL-1beta induced phosphorylation of the inhibitor of nuclear factor (NF)-kappaB (I-kappaBalpha), thus preventing its degradation. CONCLUSIONS: IL-1beta, TNFalpha, and VEGF(165) induced CAM expression in hRVECs through activation of the NF-kappaB pathway. DHA(22:6n3) inhibited cytokine induced CAM expression through suppression of NF kappaB nuclear translocation and upstream I-kappaBalpha phosphorylation and degradation. DHA(22:6n3) could be an important anti-inflammatory agent in the face of increased cytokine production and CAM expression in the diabetic retina. PMID- 16249521 TI - Antimalarials: shortages and searches. PMID- 16249522 TI - Phosphoinositide phosphatases: emerging roles as voltage sensors? AB - During a genomic survey of the transparent sea squirt (Ciona intestinalis), Murata et al. discovered a gene that encodes a protein containing homologous sequences to both a CX(5)R phosphatase and an ion channel. The authors named the novel protein, C. intestinalis voltage-sensor-containing phosphatase, Ci-VSP. The N terminus of Ci-VSP appears to function as a voltage-gated sensor; the C terminus functions as a phosphoinositide phosphatase. The authors suggest that when the N-terminal voltage sensor is activated, this in turn activates the phosphatase, which converts PI(3,4,5)P(3) to PI(4,5)P(2). Localized changes in membrane PI(4,5)P(2) levels could then serve to either positively or negatively regulate a variety of ion transporters and channels. PMID- 16249523 TI - The neuronal death protein par-4 mediates dopaminergic synaptic plasticity. AB - Par-4, discovered in a screen for genes whose expression is increased in prostate tumor cells undergoing apoptosis, participates in physiological and pathological nerve cell death. A new study, however, provides evidence for an unexpected role for Par-4 in regulating synaptic transmission in the brain: Par-4 binds to the D2 dopamine receptor (D2DR) and modulates its activity. Mice in which the function of Par-4 is disrupted exhibit impaired dopaminergic neurotransmission, resulting in a depression-like syndrome. Several other cell death-related proteins also appear to function in regulating synaptic plasticity, suggesting that a better understanding of the functions of these proteins may lead to novel therapeutic approaches for a psychiatric and neurodegenerative disorders. PMID- 16249524 TI - Serotonin 5-ht2c receptor agonists: potential for the treatment of obesity. AB - Obesity continues to be a burgeoning health problem worldwide. Before their removal from the market, fenfluramine and the more active enantiomer dexfenfluramine were considered to be among the most effective of weight loss agents. Much of the weight loss produced by fenfluramine was attributed to the direct activation of serotonin 5-HT(2C) receptors in the central nervous system via the desmethyl-metabolite of fenfluramine, norfenfluramine. Norfenfluramine, however, is non-selective, activating additional serotonin receptors, such as 5 HT(2A) and 5-HT(2B), which likely mediated the heart valve hypertrophy seen in many patients. Development of highly selective 5-HT(2C) agonists may recapitulate the clinical anti-obesity properties observed with fenfluramine while avoiding the significant cardiovascular and pulmonary side effects. PMID- 16249525 TI - Drosophila: a "model" model system to study neurodegeneration. AB - The fruit fly, Drosophila melanogaster, is a powerful model genetic organism that has been used since the turn of the previous century in the study of complex biological problems. In the last decade, numerous researchers have focused their attention on understanding neurodegenerative diseases by utilizing this model system. Numerous Drosophila mutants have been isolated that profoundly affect neural viability and integrity of the nervous system with age. Additionally, many transgenic strains have been developed as models of human disease conditions. We review the existing Drosophila neurodegenerative mutants and transgenic disease models, and discuss the role of the fruit fly in therapeutic development for neurodegenerative diseases. PMID- 16249526 TI - Neurogenic inflammation and migraine: implications for the therapeutics. AB - Significant recent advances in molecular pharmacology have elucidated the molecular pathways involved in neurogenic inflammation (NI). The release of tachykinins and endothelin-3 (ET-3) from trigeminal neurons induces dural vascular permeability and vasodilatation via activation of tachykinin receptor 1 (Tacr1) and endothelin receptor type B (Ednrb) on endothelial cells. Endothelial cell receptor stimulation results in cellular contraction, leading to plasma protein extravasation (PPE), which is the most recognized physiological hallmark of NI, and nitric oxide-induced vasodilatation. By contrast, the release of calcitonin gene-related peptide (CGRP) from trigeminal neurons--also a key physiological component of NI--does not affect vascular permeability but does induce neurogenic vasodilatation (NV) via the direct, (i.e., endothelium independent) relaxation of vascular smooth muscle. The molecular pharmacology of NI is discussed within the context of migraine research and assesses the putative role of the two key physiological components of NI (i.e., PPE and NV) in migraine pathophysiology. The data indicate that the PPE component of NI plays no significant role in migraine but that NV is likely to be involved in migraine pathophysiology. PMID- 16249527 TI - Effect of a high-carbohydrate versus a high--cis-monounsaturated fat diet on blood pressure in patients with type 2 diabetes. AB - OBJECTIVE: To investigate whether blood pressure is different in type 2 diabetic patients on a diet rich in carbohydrates versus a diet rich in cis monounsaturated fatty acids. Data on the dietary effects on these diets' glucose and lipid metabolism have been previously published. RESEARCH DESIGN AND METHODS: The study compared the effect of feeding 42 type 2 diabetic patients a carefully controlled isoenergic high-carbohydrate (high-carb; 55% energy as carbohydrate, 30% as fat, and 10% as monounsaturated fat) and high-monounsaturated fat (high mono; 45% energy as fat, 25% as monounsaturated fat, and 40% as carbohydrate) diet for 6 weeks each in a four-center, randomized, cross-over study on blood pressure. Twenty-one patients continued the diet they received during the second phase for an additional 8 weeks. RESULTS: According to repeated-measures ANOVA, blood pressure during the last 3 days of each phase was similar after 6 weeks of the high-carb and high-mono diets (systolic blood pressure: 128 +/- 16 vs. 127 +/ 15 mmHg, P = 0.9; diastolic blood pressure: 75 +/- 7 vs. 75 +/- 8 mmHg, P = 0.7). However, after 14 weeks of the high-carb diet (n = 13), there was a significant increase in blood pressure compared with 6 weeks of the high-mono diet (systolic blood pressure: 132 +/- 13 vs. 126 +/- 11 mmHg, P = 0.04; diastolic blood pressure: 83 +/- 6 vs. 76 +/- 7 mmHg, P = 0.002). After 14 weeks of the high-mono diet (n = 8), the reduction in blood pressure was not significant compared with 6 weeks of the high-carb diet (systolic blood pressure: 118 +/- 14 vs. 121 +/- 16 mmHg, P = 0.4; diastolic blood pressure: 71 +/- 8 vs. 75 +/- 10 mmHg, P = 0.3). CONCLUSION: Although the exchange of carbohydrates with monounsaturated fats may not affect blood pressure in the short term, long-term consumption of a high-carbohydrate diet may modestly raise blood pressure in type 2 diabetic patients. PMID- 16249528 TI - Incidence of type 1 and type 2 diabetes in adults aged 30-49 years: the population-based registry in the province of Turin, Italy. AB - OBJECTIVE: Incidence of type 1 diabetes is considered to be low in adults, but no study has been performed in Mediterranean countries. RESEARCH DESIGN AND METHODS: We extended the study base of the registry of the province of Turin, Italy, to subjects aged 30-49 years in the period 1999-2001 to estimate the incidences of type 1 and type 2 diabetes. Diagnosis of type 1 diabetes was based on permanent insulin treatment or a fasting C-peptide level < or =0.20 nmol/l or islet cell (ICA) or GAD (GADA) antibody positivities. RESULTS: We identified 1,135 case subjects with high completeness of ascertainment (99%), giving an incidence rate of 58.0 per 100,000 person-years (95% CI 54.7-61.5). The incidence of type 1 diabetes was 7.3 per 100,000 person-years (6.2-8.6), comparable with the rates in subjects aged 0-14 and 15-29 years (10.3 [9.5-11.2] and 6.8 [6.3-7.4]). Male subjects had a higher risk than female subjects for both type 1 (rate ratio [RR] 1.70 [95% CI 1.21-2.38]) and type 2 (2.10 [1.84-2.40]) diabetes. ICA and/or GADA positivities were found in 16% of the cohort. In logistic regression, variables independently associated with autoimmune diabetes were age 30-39 years (odds ratio [OR] 2.39 [95% CI 1.40-4.07]), fasting C-peptide <0.60 nmol/l (3.09 [1.74 5.5]), and BMI <26 kg/m2 (2.17 [1.22-3.85]). CONCLUSIONS: Risk of type 1 diabetes between age 30 and 49 years is similar to that found in the same area between age 15 and 29 years. Further studies are required to allow geographical comparisons of risks of both childhood and adulthood autoimmune diabetes, the latter being probably higher than previously believed. PMID- 16249529 TI - The effects of race and region on cardiovascular morbidity among elderly Americans with diabetes. AB - OBJECTIVE: There is conflicting evidence about whether nonwhite Americans with diabetes have an increased risk of cardiovascular disease (CVD). Because geographic region is known to influence the risk of CVD in the U.S., we sought to determine the effects of race and region on cardiovascular morbidity among elderly Americans with diabetes. RESEARCH DESIGN AND METHODS: We performed a national, retrospective, cohort study using the Medicare claims of 126,153 white and 17,962 black patients with diabetes, aged > or =65 years in 1994, who were followed through 1999 for incident acute myocardial infarction, ischemic heart disease, stroke, and heart failure. The effect of race, sex, and region on the incidence of these diseases was assessed using Cox proportional hazards regression, adjusting for baseline demographics and comorbidities. RESULTS: The incidence of any CVD ranged from 23.9/100 person-years among southern black men to 29.2/100 person-years among non-southern black women. The risk of CVD was lower among southern black men (hazard ratio 0.87 [95% CI 0.82-0.92]) and women (0.95 [0.91-0.99]) than their southern white counterparts. In the three other U.S. regions combined (northeast, midwest, and west), black men had a similar risk for CVD (1.01 [0.95-1.07]), and black women had a greater risk (1.10 [1.05 1.16]) than non-southern white men and women, respectively. CONCLUSIONS: Among elderly Americans with diabetes, the incidence of CVD is unlikely to differ a great deal between whites and blacks. Residence in the South seems to confer a modest benefit for elderly black people with diabetes. PMID- 16249531 TI - Restless legs syndrome and quality of sleep in type 2 diabetes. AB - OBJECTIVE: To investigate the presence of restless legs syndrome (RLS) and the quality of sleep in a population of type 2 diabetic patients. RESEARCH DESIGN AND METHODS: The study population was composed of 100 consecutive patients regularly attending a diabetes clinic at the University Hospital of the Federal University of Ceara. The subjects' quality of sleep was assessed by the Pittsburgh Sleep Quality Index, and excessive daytime sleepiness (EDS) was measured by the Epworth Sleepiness Scale. The RLS was diagnosed using the four minimum criteria defined by the International Restless Legs Syndrome Study Group. Other relevant clinical and laboratory parameters were obtained by interview and chart review. RESULTS: RLS was found in 27% of patients. Poor sleep quality was present in 45% of cases and was associated with age (P = 0.04), peripheral neuropathy (P = 0.001), and RLS (P = 0.000). EDS was found in 26% of patients. Logistic regression analysis revealed an association between RLS and peripheral neuropathy (odds ratio 12.85 [95% CI 2.83-58.40], P = 0.001). CONCLUSIONS: RLS is common in type 2 diabetic patients and can be a major cause of sleep disruption in these patients. PMID- 16249530 TI - The relation of fasting and 2-h postchallenge plasma glucose concentrations to mortality: data from the Baltimore Longitudinal Study of Aging with a critical review of the literature. AB - OBJECTIVE: Under the auspices of the National Institutes of Health, American Diabetes Association, and World Health Organization, expert committees lowered the fasting plasma glucose (FPG) concentration diagnostic for diabetes from 7.8 to 7.0 mmol/l and defined 6.1-6.9 mmol/l as impaired fasting glucose (IFG) and <6.1 mmol/l as normal fasting glucose. In 2003, IFG was lowered to 5.6-6.9 mmol/l and normal fasting glucose to <5.6 mmol/l. Reports of the relationship between glucose concentration and all-cause mortality have been inconsistent. It is not known if the 2-h plasma glucose (2hPG) concentration from an oral glucose tolerance test (OGTT) adds to the predictive power of FPG. RESEARCH DESIGN AND METHODS: We followed 1,236 men for an average of 13.4 years to determine the relationship between both FPG and 2hPG and all-cause mortality. RESULTS: Risk for mortality did not increase until the FPG exceeded 6.1 mmol/l. Risk increased by approximately 40% in the 6.1-6.9 mmol/l range and doubled when FPG ranged from 7.0 to 7.7 mmol/l. A combination of the 2hPG and FPG allowed better estimation of risk than the FPG alone. Within any category of FPG, risk generally increased as the 2hPG increased, and within any category of 2hPG, risk generally increased as the FPG increased. CONCLUSIONS: These data support the decision to lower the FPG diagnostic for diabetes from 7.8 to 7.0 mmol/l, show that both IFG and impaired glucose tolerance have risks between the normal and diabetic ranges, and show that the OGTT adds predictive power to that of FPG alone and should not be abandoned. The lowering of IFG to 5.6 mmol/l from 6.1 mmol/l, at least for mortality, is, however, not supported by our results. PMID- 16249532 TI - Longitudinal assessment of quality of life in patients with type 2 diabetes and self-reported erectile dysfunction. AB - OBJECTIVE: In the context of the QuED (Quality of Care and Outcomes in Type 2 Diabetes) project, we evaluated the longitudinal changes over 3 years in quality of life (QoL) in patients with type 2 diabetes according to the presence or the development of erectile dysfunction (ED). RESEARCH DESIGN AND METHODS: Patients were requested to fill in a questionnaire investigating the presence of ED and QoL (SF-36 Health Survey, depression symptoms [Center for Epidemiologic Studies Depression], and quality of sexual life) every 6 months for 3 years. The analyses were based on multilevel models, adjusted for patient clinical and sociodemographic characteristics. RESULTS: The study involved 1,456 patients, of whom 34% reported frequent erectile problems at baseline; 192 developed ED during the follow-up. No changes in QoL measures were detected in patients without ED; in those with ED at baseline, a worsening in all SF-36 scales was observed, reaching statistical significance for physical functioning (P = 0.03). Among patients who developed ED during the study, a deterioration in all SF-36 dimensions and a worsening in depressive symptoms preceded the development of ED. The onset of ED was associated with a further marked worsening in physical functioning (P = 0.0008), general health perception (P = 0.02), and social functioning (P = 0.04) on SF-36 subscales, as well as in the summary physical and mental components scores (P = 0.04 and P = 0.07, respectively). The development of ED was also associated with a highly significant increase in depressive symptoms (P = 0.001) and a marked decrease in quality of sexual life (P < 0.0001). CONCLUSIONS: This longitudinal study documents for the first time the impact of ED onset on several aspects of QoL in patients with type 2 diabetes. The study also shows that QoL tended to further decrease during 3 years in patients with ED at baseline but not in those without this condition. PMID- 16249533 TI - Does diabetes care compete with the provision of women's preventive care services? AB - OBJECTIVE: Care for chronic diseases may compete with preventive health care. To test this hypothesis, we examined the association between diabetes-related processes of care and preventive care in women. RESEARCH DESIGN AND METHODS: Using data from a prospective cohort study of diabetes care in managed care settings, we reviewed the care 540 diabetic women received from 355 primary care providers within 14 provider groups from one health plan. Of the 540 women, 278 were eligible to receive mammograms and 314 were eligible to receive Pap smears. Mammography performance was measured as at least one mammogram over a 2-year period and Pap performance was measured as at least one Pap smear over a 3-year period. To assess the association between diabetes-related processes of care and preventive services, we used hierarchical logistic regression models, accounted for clustering within provider groups, and adjusted for patient age, race, income and education level, diabetes treatment and duration, and health status, as well as physician age, sex, years of practice, and specialty. Diabetes-related processes of care were defined as dilated retinal examinations, urine microalbumin/protein testing, foot examinations, lipid and HbA(1c) testing, recommendations to take aspirin, and influenza vaccinations received over a 1 year period. RESULTS: In this cohort, 73% of eligible women received mammograms and 56% received Pap smears. After adjustment of models, better diabetes-related processes of care, better health status, and non-Medicaid insurance were associated with mammography performance. Better diabetes-related processes of care, younger patient age, and any visit to a gynecologist were associated with Pap performance. CONCLUSIONS: Better processes of diabetes care were associated with better women's preventive health care. Diabetes management did not compete with sex-specific screening. PMID- 16249534 TI - Postpartum physical activity and related psychosocial factors among women with recent gestational diabetes mellitus. AB - OBJECTIVE: In this study, we examined patterns of postpartum physical activity among women with recent gestational diabetes mellitus (GDM) and psychosocial factors related to this behavior that could be addressed in diabetes prevention interventions. RESEARCH DESIGN AND METHODS: A random sample of women who had attended diabetes clinics in Sydney, Australia, in the past 6-24 months for treatment of GDM were surveyed by telephone. Variables measured included physical activity behaviors, self-efficacy, social support, and barriers to participation. RESULTS: Of 226 women who completed the survey (mean age 33.4 years), 26.5% were classified as sedentary, and only 33.6% reported sufficient physical activity as recommended by health authorities. Walking was the most popular physical activity, and most women reported no other moderate- or vigorous-intensity activity. Lack of assistance with child care (49.1%) and insufficient time (37.6%) were the most common barriers to physical activity. The type of social support most often reported was verbal encouragement (39.1%), with more than half of the women never receiving assistance with housework or others exercising with them. Self-efficacy for physical activity was lowest when women were under time pressure or tired. Multivariate analyses showed that sufficient physical activity was associated with high social support (odds ratio 2.5 [95% CI 1.21-3.79]) and high self-efficacy (2.09 [1.06-3.20]). CONCLUSIONS: The prevalence of sufficient physical activity was found to be low and strongly related to social support and self-efficacy. This is an important group to whom diabetes prevention strategies can be targeted. PMID- 16249535 TI - Use of the Patient Assessment of Chronic Illness Care (PACIC) with diabetic patients: relationship to patient characteristics, receipt of care, and self management. AB - OBJECTIVE: There is a dearth of information on the extent to which diabetic patients receive care congruent with the chronic care model (CCM) and evidence based behavioral counseling. This study evaluates a new instrument to fill this gap. RESEARCH DESIGN AND METHODS: A heterogeneous sample of 363 type 2 diabetic patients completed the original Patient Assessment of Chronic Illness Care (PACIC), along with additional items that allowed it to be scored according to the "5As" (ask, advise, agree, assist, and arrange) model of behavioral counseling. We evaluated relationships between survey scores and patient characteristics, quality of diabetes care, and self-management. RESULTS: Findings replicated those of the initial PACIC validation study but with a much larger sample of diabetic patients and more Latinos. Areas of CCM activities reported least often were goal setting/intervention tailoring and follow-up/coordination. The 5As scoring revealed that patients were least likely to receive assistance with problem solving and arrangement of follow-up support. Few demographic or medical characteristics were related to PACIC or 5As scores, but survey scores were significantly related to quality of diabetes care received and level of physical activity. CONCLUSIONS: The PACIC and the new 5As scoring method appear useful for diabetic patients. Its use is encouraged in future research and quality improvement studies. PMID- 16249536 TI - The impact of diabetes on employment and work productivity. AB - OBJECTIVE: The purpose of this study was to longitudinally examine the effect of diabetes on labor market outcomes. RESEARCH DESIGN AND METHODS: Using secondary data from the first two waves (1992 and 1994) of the Health and Retirement Study, we identified 7,055 employed respondents (51-61 years of age), 490 of whom reported having diabetes in wave 1. We estimated the effect of diabetes in wave 1 on the probability of working in wave 2 using probit regression. For those working in wave 2, we modeled the relationships between diabetic status in wave 1 and the change in hours worked and work-loss days using ordinary least-squares regressions and modeled the presence of health-related work limitations using probit regression. All models control for health status and job characteristics and are estimated separately by sex. RESULTS: Among individuals with diabetes, the absolute probability of working was 4.4 percentage points less for women and 7.1 percentage points less for men relative to that of their counterparts without diabetes. Change in weekly hours worked was not statistically significantly associated with diabetes. Women with diabetes had 2 more work-loss days per year compared with women without diabetes. Compared with individuals without diabetes, men and women with diabetes were 5.4 and 6 percentage points (absolute increase), respectively, more likely to have work limitations. CONCLUSIONS: This article provides evidence that diabetes affects patients, employers, and society not only by reducing employment but also by contributing to work loss and health-related work limitations for those who remain employed. PMID- 16249537 TI - The association of comorbid depression with mortality in patients with type 2 diabetes. AB - OBJECTIVE: We assessed whether patients with comorbid minor and major depression and type 2 diabetes had a higher mortality rate over a 3-year period compared with patients with diabetes alone. RESEARCH DESIGN AND METHODS: In a large health maintenance organization (HMO), 4,154 patients with type 2 diabetes were surveyed and followed for up to 3 years. Patients initially filled out a written questionnaire, and HMO-automated diagnostic, laboratory, and pharmacy data and Washington State mortality data were collected to assess diabetes complications and deaths. Cox proportional hazards regression models were used to calculate adjusted hazard ratios of death for each group compared with the reference group. RESULTS: There were 275 (8.3%) deaths in 3,303 patients without depression compared with 48 (13.6%) deaths in 354 patients with minor depression and 59 (11.9%) deaths among 497 patients with major depression. A proportional hazards model with adjustment for age, sex, race/ethnicity, and educational attainment found that compared with the nondepressed group, minor depression was associated with a 1.67-fold increase in mortality (P = 0.003), and major depression was associated with a 2.30-fold increase (P < 0.0001). In a second model that controlled for multiple potential mediators, both minor and major depression remained significant predictors of mortality. CONCLUSIONS: Among patients with diabetes, both minor and major depression are strongly associated with increased mortality. Further research will be necessary to disentangle causal relationships among depression, behavioral risk factors (adherence to medical regimens), diabetes complications, and mortality. PMID- 16249538 TI - Resistance to insulin therapy among patients and providers: results of the cross national Diabetes Attitudes, Wishes, and Needs (DAWN) study. AB - OBJECTIVE: To examine the correlates of patient and provider attitudes toward insulin therapy. RESEARCH DESIGN AND METHODS: Data are from surveys of patients with type 2 diabetes not taking insulin (n = 2,061) and diabetes care providers (nurses = 1,109; physicians = 2,681) in 13 countries in Asia, Australia, Europe, and North America. Multiple regression analysis is used to identify correlates of attitudes toward insulin therapy among patients, physicians, and nurses. RESULTS: Patient and provider attitudes differ significantly across countries, controlling for individual characteristics. Patients rate the clinical efficacy of insulin as low and would blame themselves if they had to start insulin therapy. Self-blame is significantly lower among those who have better diet and exercise adherence and less diabetes-related distress. Patients who are not managing their diabetes well (poor perceived control, more complications, and diabetes-related distress) are significantly more likely to see insulin therapy as potentially beneficial. Most nurses and general practitioners (50-55%) delay insulin therapy until absolutely necessary, but specialists and opinion leaders are less likely to do so. Delay of insulin therapy is significantly less likely when physicians and nurses see their patients as more adherent to medication or appointment regimens, view insulin as more efficacious, and when they are less likely to delay oral diabetes medications. CONCLUSIONS: Patient and provider resistance to insulin therapy is substantial, and for providers it is part of a larger pattern of reluctance to prescribe blood glucose-lowering medication. Interventions to facilitate timely initiation of insulin therapy will need to address factors associated with this resistance. PMID- 16249539 TI - Association among individual deprivation, glycemic control, and diabetes complications: the EPICES score. AB - OBJECTIVE: Previous studies have related poor glycemic control and/or some diabetes complications to low socioeconomic status. Some aspects of socioeconomic status have not been assessed in these studies. In the present study, we used an individual index of deprivation, the Evaluation de la Precarite et des Inegalites de sante dans les Centres d'Examens de Sante (Evaluation of Precarity and Inequalities in Health Examination Centers [EPICES]) score, to determine the relationship among glycemic control, diabetes complications, and individual conditions of deprivation. RESEARCH DESIGN AND METHODS: We conducted a cross sectional prevalence study in 135 consecutive diabetic patients (age 59.41 +/- 13.2 years [mean +/- SD]) admitted in the hospitalization unit of a French endocrine department. Individual deprivation was assessed by the EPICES score, calculated from 11 socioeconomic questions. Glycemic control, lipid levels, blood pressure, retinopathy, neuropathy, and nephropathy were assessed. RESULTS: HbA(1c) level was significantly correlated with the EPICES score (r = 0.366, P < 0.001). The more deprived patients were more likely than the less deprived patients to have poor glycemic control (beta = 1.984 [SE 0.477], P < 0.001), neuropathy (odds ratio 2.39 [95% CI 1.05-5.43], P = 0.037), retinopathy (3.66 [1.39-9.64], P = 0.009), and being less often admitted for 1-day hospitalization (0.32 [0.14-0.74], P = 0.008). No significant relationship was observed with either nephropathy or cardiovascular risk factors. CONCLUSIONS: Deprivation status is associated with poor metabolic control and more frequent microvascular complications, i.e., retinopathy and neuropathy. The medical and economic burden of deprived patients is high. PMID- 16249540 TI - The effect of ruboxistaurin on nephropathy in type 2 diabetes. AB - OBJECTIVE: Ruboxistaurin selectively inhibits protein kinase C-beta and ameliorates kidney disease in animal models of diabetes. The purpose of this study was to evaluate the effects of ruboxistaurin on diabetic nephropathy in humans. RESEARCH DESIGN AND METHODS: A randomized, double-blind, placebo controlled, multicenter, pilot study was performed to evaluate the effects of 32 mg/day ruboxistaurin for 1 year in persons (n = 123) with type 2 diabetes and persistent albuminuria (albumin-to-creatinine ratio [ACR] 200-2,000 mg/g), despite therapy with renin-angiotensin system inhibitors. The primary end point was a change in the ACR. Estimated glomerular filtration rate (eGFR) (four component equation from the Modification of Diet in Renal Disease study) was also calculated. RESULTS: At baseline, urinary ACR was 764 +/- 427 mg/g (means +/- SD), and eGFR was 70 +/- 24 ml/min per 1.73 m2. Systolic and diastolic blood pressures were 135 +/- 14 and 75 +/- 9 mmHg, respectively. HbA(1c) was 8.0 +/- 1.2%. After 1 year, urinary ACR decreased significantly (-24 +/- 9%) in participants treated with ruboxistaurin (P = 0.020) and nonsignificantly (-9 +/- 11%) in the placebo group (P = 0.430). The ACR-lowering effect of ruboxistaurin appeared by 1 month. eGFR did not decline significantly in the ruboxistaurin group (-2.5 +/- 1.9 ml/min per 1.73 m2) (P = 0.185), whereas the placebo group lost significant eGFR over 1 year (-4.8 +/- 1.8 ml/min per 1.73 m2) (P = 0.009). Between-group differences for changes in ACR and eGFR were not statistically significant, but this pilot study was underpowered to determine such differences. CONCLUSIONS: In persons with type 2 diabetes and nephropathy, treatment with ruboxistaurin reduced albuminuria and maintained eGFR over 1 year. Ruboxistaurin may add benefit to established therapies for diabetic nephropathy. PMID- 16249541 TI - Increasing the decrement in insulin secretion improves glucagon responses to hypoglycemia in advanced type 2 diabetes. AB - OBJECTIVE: In advanced beta-cell failure, counterregulatory glucagon responses may be impaired due to a reduced decrement in insulin secretion during the development of hypoglycemia. The present studies were therefore undertaken to test the hypothesis that these may be improved by increasing this decrement in insulin secretion. RESEARCH DESIGN AND METHODS: Twelve subjects with type 2 diabetes who have been insulin requiring were studied as a model of advanced beta cell failure. Glucagon responses were examined during a 90-min hypoglycemic clamp (approximately 2.8 mmol/l) on two separate occasions. On one occasion, tolbutamide was infused for 2 h before the clamp so that the decrement in insulin secretion during the induction of hypoglycemia would be increased. On the other occasion, normal saline was infused as a control. RESULTS: Before the hypoglycemic clamp, infusion of tolbutamide increased insulin secretion approximately 1.9-fold (P < 0.001). However, during hypoglycemia, insulin secretion decreased to similar rates on both occasions (P = 0.31) so that its decrement was approximately twofold greater following the tolbutamide infusion (1.63 +/- 0.20 vs. 0.81 +/- 0.17 pmol x kg(-1) x min(-1), P < 0.001). This was associated with more than twofold-greater glucagon responses (42 +/- 11 vs. 19 +/ 8 ng/l, P < 0.002) during the hypoglycemic clamp but unaltered glucagon responses to intravenous arginine immediately thereafter (449 +/- 50 vs. 453 +/- 50 ng/l, P = 0.78). CONCLUSIONS: Increasing the decrement in insulin secretion during the development of hypoglycemia improves counterregulatory glucagon responses in advanced beta-cell failure. These findings further support the concept that the impaired counterregulatory glucagon responses in advanced beta cell failure may at least partially be due to a reduced decrement in insulin secretion. PMID- 16249542 TI - A randomized controlled trial of the effect of real-time telemedicine support on glycemic control in young adults with type 1 diabetes (ISRCTN 46889446). AB - OBJECTIVE: To determine whether a system of telemedicine support can improve glycemic control in type 1 diabetes. RESEARCH DESIGN AND METHODS: A 9-month randomized trial compared glucose self-monitoring real-time result transmission and feedback of results for the previous 24 h in the control group with real-time graphical phone-based feedback for the previous 2 weeks together with nurse initiated support using a web-based graphical analysis of glucose self-monitoring results in the intervention group. All patients aged 18-30 years with HbA(1c) (A1C) levels of 8-11% were eligible for inclusion. RESULTS: A total of 93 patients (55 men) with mean diabetes duration (means +/- SD) 12.1 +/- 6.7 years were recruited from a young adult clinic. In total, the intervention and control groups transmitted 29,765 and 21,400 results, respectively. The corresponding median blood glucose levels were 8.9 mmol/l (interquartile range 5.4-13.5) and 10.3 mmol/l (6.5-14.4) (P < 0.0001). There was a reduction in A1C in the intervention group after 9 months from 9.2 +/- 1.1 to 8.6 +/- 1.4% (difference 0.6% [95% CI 0.3-1.0]) and a reduction in A1C in the control group from 9.3 +/- 1.5 to 8.9 +/- 1.4% (difference 0.4% [0.03-0.7]). This difference in change in A1C between groups was not statistically significant (0.2% [-0.2 to 0.7, P = 0.3). CONCLUSIONS: Real-time telemedicine transmission and feedback of information about blood glucose results with nurse support is feasible and acceptable to patients, but to significantly improve glycemic control, access to real-time decision support for medication dosing and changes in diet and exercise may be required. PMID- 16249543 TI - Laparoscopic gastric banding prevents type 2 diabetes and arterial hypertension and induces their remission in morbid obesity: a 4-year case-controlled study. AB - OBJECTIVE: Lifestyle modifications and pharmacological interventions can prevent type 2 diabetes in obese subjects with impaired glucose tolerance. The aim of this study was to compare laparoscopic adjustable gastric banding (LAGB) and conventional diet (No-LAGB) in the prevention (primary intervention study; 56 vs. 29 patients) and remission (secondary intervention study; 17 vs. 20 patients) of type 2 diabetes and hypertension in grade 3 obesity in a 4-year study. RESEARCH DESIGN AND METHODS: The subjects (n = 122; age 48.5 +/- 1.05 years; BMI 45.7 +/- 0.67 kg/m2) underwent a diagnostic workup, including psychological and psychiatric assessments, in preparation for the LAGB procedure. Of the 122 subjects, 73 had the surgery (LAGB group). The control group (No-LAGB group) consisted of the 49 subjects who refused the surgery but agreed to be followed up; 6 of these subjects dropped out by the 2nd year of the study, so that the final number of patients was 73 and 43 in the LAGB and No-LAGB groups, respectively. All patients had a yearly visit and oral glucose tolerance test. RESULTS: From baseline to the end of the 4-year follow-up, BMI decreased from 45.9 +/- 0.89 at baseline to 37.7 +/- 0.71 kg/m2 in the LAGB group and remained steady in the No-LAGB group (from 45.2 +/- 1.04 to 46.5 +/- 1.37 kg/m2), with no significant differences between the primary and secondary intervention groups. In the primary intervention study, five of the No-LAGB subjects (17.2%) and none of the LAGB subjects (0.0%; P = 0.0001) progressed to type 2 diabetes; in the secondary intervention study, type 2 diabetes remitted in one No-LAGB patient (4.0%) and seven LAGB patients (45.0%; P = 0.0052). Hypertension occurred in 11 No-LAGB patients (25.6%) and 1 LAGB patient (1.4%; P = 0.0001) and remitted in 1 No-LAGB (2.3%) and 15 LAGB patients (20.5%; P = 0.0001). A study of body mass composition revealed a significant reduction of fat mass and a transitory, but not significant, decrease of fat-free mass in LAGB patients. CONCLUSIONS: In morbid obesity, sustained and long-lasting weight loss obtained through LAGB prevents the occurrence of type 2 diabetes and hypertension and decreases the prevalence of these disorders. PMID- 16249544 TI - The impact of insulin resistance on proinsulin secretion in pregnancy: hyperproinsulinemia is not a feature of gestational diabetes. AB - OBJECTIVE: Excessive secretion of the insulin precursor proinsulin, as manifested by an increased serum proinsulin-to-insulin ratio, has been associated with beta cell dysfunction. In women with gestational diabetes mellitus (GDM), previous studies of the proinsulin-to-insulin ratio have yielded conflicting results, despite the presence of beta-cell dysfunction. The interpretation of the proinsulin-to-insulin ratio, however, may be confounded by the variable effects of hepatic insulin extraction. Thus, we sought to determine whether GDM is characterized by relative hyperproinsulinemia as measured by the proinsulin-to-C peptide ratio, an alternate measure of proinsulin secretion that is not affected by hepatic insulin extraction. RESEARCH DESIGN AND METHODS: Serum proinsulin, C peptide, and insulin were measured in a cross-sectional study of 180 women undergoing oral glucose tolerance tests (OGTTs) in the late second or early third trimester. Based on the OGTT, participants were stratified into three groups: 1) normal glucose tolerance (NGT; n = 93), 2) impaired glucose tolerance (IGT; n = 39), and 3) GDM (n = 48). Insulin sensitivity (IS) was measured using the IS(OGTT) index of Matsuda and DeFronzo, which has been previously validated in pregnant women. RESULTS: There were no significant differences in mean fasting proinsulin-to-C-peptide ratio between the three glucose tolerance groups (NGT, 0.024; IGT, 0.022; GDM, 0.019; P = 0.4). Furthermore, adjustment for age, weeks' gestation, prepregnancy BMI, ethnicity, previous GDM, and family history of diabetes did not reveal any association between the proinsulin-to-C-peptide ratio and glucose tolerance status. Using Spearman univariate correlation analysis, fasting proinsulin-to-C-peptide ratio was significantly correlated with IS(OGTT) (r = 0.29, P < 0.0001) and inversely related to the homeostasis model assessment of insulin resistance (r = -0.36, P < 0.0001) and prepregnancy BMI (r = -0.23, P < 0.005). On multiple linear regression analysis, IS(OGTT) emerged as the strongest independent correlate of the dependent variable proinsulin-to-C-peptide ratio. Furthermore, after adjustment for potential covariates, a stepwise decrease in proinsulin-to-C-peptide ratio was observed per decreasing tertile of IS(OGTT) (trend P = 0.0019), consistent with enhanced efficiency of proinsulin processing (i.e., reduced proinsulin-to-C-peptide ratio) as insulin resistance increases. CONCLUSIONS: GDM is not independently associated with hyperproinsulinemia as measured by the proinsulin-to-C-peptide ratio. Instead, in pregnant women, increased insulin resistance is associated with decreased proinsulin-to-C-peptide ratio, independently of glucose tolerance status. These data suggest that relative proinsulin secretion in late pregnancy is primarily related to insulin resistance and does not necessarily reflect beta-cell function. PMID- 16249545 TI - Decreased endogenous secretory advanced glycation end product receptor in type 1 diabetic patients: its possible association with diabetic vascular complications. AB - OBJECTIVE: The binding of advanced glycation end products (AGEs) to their receptor (RAGE) plays an important role in the development of diabetic vascular complications. In the present study, we examined circulating endogenous secretory RAGE (esRAGE) levels in subjects with type 1 diabetes and explored the possible association between esRAGE levels and the severity of diabetic vascular complications. RESEARCH DESIGN AND METHODS: Circulating esRAGE levels in serum were examined in 67 Japanese type 1 diabetic patients (22 men and 45 women, age 24.0 +/- 4.4 years [means +/- SD]) and 23 age-matched healthy nondiabetic subjects (10 men and 13 women aged 24.9 +/- 1.4 years). Daily urinary albumin excretion, the presence of retinopathy, and intima-media thickness (IMT) of the carotid artery were also evaluated. We further explored the association between esRAGE levels and severity of diabetic vascular complications. RESULTS: Circulating esRAGE levels were significantly lower in subjects with type 1 diabetes than in nondiabetic subjects (0.266 +/- 0.089 vs. 0.436 +/- 0.121 ng/ml, respectively, P < 0.0001) and was inversely correlated with HbA(1c) (A1C) levels (r = -0.614, P < 0.0001). In addition, multivariate regression analysis demonstrated that A1C was an independent risk factor for a low esRAGE value. Furthermore, circulating esRAGE levels were inversely correlated with carotid IMT (r = -0.325, P = 0.0017) and was one of the independent risk factors for IMT thickening. Furthermore, there was a significant difference (P = 0.0124) in esRAGE levels between patients without retinopathy (0.286 +/- 0.092 ng/ml) and those with retinopathy (0.230 +/- 0.074 ng/ml). CONCLUSIONS: Circulating esRAGE levels were significantly lower in type 1 diabetic patients than in nondiabetic subjects and were inversely associated with the severity of some diabetic vascular complications. PMID- 16249546 TI - Predictive value of silent myocardial ischemia for cardiac events in diabetic patients: influence of age in a French multicenter study. AB - OBJECTIVE: Silent myocardial ischemia (SMI) in asymptomatic subjects with no history of myocardial infarction or angina is a frequent condition in diabetic patients. The aim of the study was to examine the predictive value of SMI for cardiac events in a multicenter cohort and to determine whether this value is higher in patients with a particular clinical profile. RESEARCH DESIGN AND METHODS: A total of 370 asymptomatic diabetic patients with at least two additional cardiovascular risk factors was recruited in four departments of diabetology. SMI was assessed by either exercise or dipyridamole single-photon emission-computed tomography myocardial perfusion imaging with thallium-201. If dipyridamole stress was used, an electrocardiogram stress test was performed separately on another day. Follow-up duration was 3-89 months (38 +/- 23 months). RESULTS: There was evidence of SMI in 131 patients (35.4%) on at least one positive noninvasive test. The patients with SMI were significantly older and had significantly higher serum triglycerides and lower HDL cholesterol levels. Cardiac events occurred in 53 patients (14.3%). Major cardiac events (death or myocardial infarction) occurred in 38 patients (10%) and other events (unstable angina, heart failure, or coronary revascularization) occurred in 15 patients. The patients who had cardiac events were older and had higher serum triglyceride levels at baseline. There was a significant association between SMI and cardiac events (hazard ratio 2.79 [95% CI 1.54-5.04]) and in particular major cardiac events (3 [1.53-5.87]). In the patients >60 years of age, the prevalence of SMI was higher (43.4 vs. 30.2% in those <60 years). SMI was associated with a significant risk of cardiac events (2.89 [1.31-6.39]) and in particular major cardiac events (3.66 [1.36-9.87]) for the patients >60 years old but not for those <60 years old. CONCLUSIONS: In asymptomatic diabetic patients with additional cardiovascular risk factors, SMI is a potent predictor of cardiac events and should be assessed preferably in the patients >60 years of age. PMID- 16249547 TI - Effect of pitavastatin on urinary liver-type fatty acid-binding protein levels in patients with early diabetic nephropathy. AB - OBJECTIVE: Liver-type fatty acid-binding protein (l-FABP) is expressed in renal proximal tubules and is reported to be a useful marker for progression of chronic glomerulonephritis. The aim of this study was to determine whether urinary l-FABP levels are altered at various stages of diabetic nephropathy and whether pitavastatin affects urinary l-FABP levels in early diabetic nephropathy. RESEARCH DESIGN AND METHODS: Fifty-eight patients with type 2 diabetes (34 men and 24 women, median age 52 years) and 20 healthy, age-matched subjects (group E) were recruited for the study. The diabetic patients included 12 patients without nephropathy (group A), 20 patients with microalbuminuria (group B), 14 patients with macroalbuminuria and normal renal function (group C), and 12 patients with chronic renal failure but not undergoing hemodialysis (blood creatinine >1.2 mg/dl; mean 2.5 mg/dl, group D). Twenty group B patients were randomly assigned to receive 1 mg/day pitavastatin (10 patients, group B1) or placebo (10 patients, group B2). Treatment was continued for 12 months. Urinary l-FABP levels were measured by enzyme-linked immunosorbent assay. Urinary 8-hydroxydeoxyguanosine and serum free fatty acids (FFAs) were also measured in group B. RESULTS: Urinary l-FABP levels in groups A-D were 6.2 +/- 4.6 microg/g creatinine, 19.6 +/- 13.5 microg/g creatinine, 26.8 +/- 20.4 microg/g creatinine, and 52.4 +/- 46.8 microg/g creatinine, respectively. Urinary l-FABP levels in groups B-D were significantly higher than those in healthy subjects (group E, 5.8 +/- 4.0 microg/g creatinine) (group B, P < 0.05; group C, P < 0.01; group D, P < 0.01). In group B1, urinary albumin excretion (UAE) and urinary l-FABP levels were decreased after pitavastatin treatment (UAE before, 110 +/- 74 microg/min; 6 months, 88 +/- 60 microg/min, P < 0.05; 12 months, 58 +/- 32 microg/min, P < 0.01; l-FABP before, 18.6 +/- 12.5 microg/g creatinine; 6 months, 12.2 +/- 8.8 microg/g creatinine, P < 0.05; 12 months, 8.8 +/- 6.4 microg/g creatinine, P < 0.01). In group B2, UAE and l-FABP levels showed little change during the experimental period. In group B1, urinary 8-hydroxydeoxyguanosine was decreased 12 months after pitavastatin treatment (before 32.5 +/- 19.5 ng/mg creatinine, after 18.8 +/- 14.5 ng/mg creatinine, P < 0.01), but in group B2, these showed little difference during the experimental period. In both groups B1 and B2, serum FFAs showed little difference during the experimental period. CONCLUSIONS: Urinary l-FABP levels appear to be associated with the progression of diabetic nephropathy, and pitavastatin may be effective in ameliorating tubulointerstitial damage in early diabetic nephropathy. PMID- 16249548 TI - Development, progression, and regression of microalbuminuria in Japanese patients with type 2 diabetes under tight glycemic and blood pressure control: the Kashiwa study. AB - OBJECTIVE: The goal of this study was to know the fate of albuminuria in Japanese patients with type 2 diabetes under tight blood pressure and glycemic control. RESEARCH DESIGN AND METHODS: Patients having normoalbuminuria (urinary albumin excretion <30 mg/g creatinine, n = 179) or microalbuminuria (albumin excretion 30 299 mg/g creatinine, n = 94) at baseline have been followed up for 8 years: ratio of men to women was 160/113, the mean age was 58 years, pretreatment HbA(1c) (A1C) was 8.8%, and blood pressure was 136/76 mmHg. A1C <6.5% and blood pressure <130/80 mmHg were targeted, and the A1C of 6.5 +/- 0.7% (mean +/- SD) and blood pressure of 127 +/- 11/72 +/- 6 mmHg have been maintained during the 8 years. Development of microalbuminuria or macroalbuminuria (albumin excretion > or =300 mg/g creatinine) in initially normoalbuminuric patients and progression to macroalbuminuria or regression to normoalbuminuria in initially microalbuminuric patients were assessed at year 8. RESULTS: Development occurred in 27 (15%) of the normoalbuminuric patients and progression and regression in 16 (17%) and 20 (21%), respectively, of the microalbuminuric patients. Significant independent relationships existed between development and higher achieved mean systolic blood pressure (SBP) and regression and lower achieved mean SBP. In the patients with achieved mean SBP <120 mmHg, development was 3%, progression was 11%, and regression was 44% during 8 years. Prediction for nephropathy by blood pressure and glycemia alone was limited. Nevertheless, albumin excretion at year 8 was positively correlated with achieved mean SBP and baseline albuminuria. CONCLUSIONS: Development and progression were low and regression was high with SBP of 120 mmHg, provided A1C was maintained at 6.5%. PMID- 16249549 TI - Sleep disturbances in midlife unrelated to 32-year diabetes incidence: the prospective population study of women in Gothenburg. AB - OBJECTIVE: To study the relation between diabetes incidence and sleep problems in a population-based sample of women followed for 32 years. RESEARCH DESIGN AND METHODS: The researchers conducted a prospective population study initiated in 1968-1969, with follow-ups in 1974-1975, 1980-1981, 1992-1993, and 2000-2001 in Gothenburg, Sweden. A total of 1,462 women born in 1908, 1914, 1918, 1922, and 1930, representative of women of the same ages in the general population, initially participated (90% participation rate). Reported sleep duration, sleep problems, and use of sleeping medication were related to incident diabetes from 1968 to 2000. Associations between sleep problems and diabetes were corrected for waist-to-hip ratio (WHR), BMI, subscapular skinfold, fasting blood glucose and serum lipid concentrations, blood pressure, heart rate, smoking, physical activity, education, and socioeconomic status. Additionally, associations between BMI, WHR, and sleep problems were examined. RESULTS: Over 32 years, 126 women (8.7%) developed diabetes. Associations between diabetes and initial sleep problems were tested in a Cox regression analysis, taking into consideration factors associated (P < 0.1) with diabetes. Sleep problems in 1968 did not increase risk of developing diabetes during the following 32 years. Obesity, particularly centralized, was associated with sleep problems. CONCLUSIONS: No association between sleep problems and developing diabetes was seen in this 32 year follow-up of middle-aged women. Obesity, on the other hand, known to cause increased risk of diabetes, was associated with current sleep problems. PMID- 16249550 TI - Prevalence of the metabolic syndrome defined by the International Diabetes Federation among adults in the U.S. AB - OBJECTIVE: The International Diabetes Federation (IDF) has proposed a new definition of the metabolic syndrome that emphasizes central adiposity as determined by ethnic group-specific thresholds of waist circumference. The objective of this study was to estimate the prevalence of this syndrome using the IDF definition among U.S. adults and to compare it with the prevalence estimated using the definition of the National Cholesterol Education Program (NCEP). RESEARCH DESIGN AND METHODS: A total of 3,601 men and women aged > or =20 years from the National Health and Nutrition Examination Survey 1999-2002 were included in the analyses. RESULTS: Based on the NCEP definition, the unadjusted prevalence of the metabolic syndrome was 34.5 +/- 0.9% (percent +/- SE) among all participants, 33.7 +/- 1.6% among men, and 35.4 +/- 1.2% among women. Based on the IDF definition, the unadjusted prevalence of the metabolic syndrome was 39.0 +/- 1.1% among all participants, 39.9 +/- 1.7% among men, and 38.1 +/- 1.2% among women. The IDF definition led to higher estimates of prevalence in all of the demographic groups, especially among Mexican-American men. The two definitions similarly classified approximately 93% of the participants as having or not having the metabolic syndrome. CONCLUSIONS: In the U.S., the use of the IDF definition of the metabolic syndrome leads to a higher prevalence estimate of the metabolic syndrome than the estimate based on the NCEP definition. PMID- 16249551 TI - Elevated homocysteine as a risk factor for the development of diabetes in women with a previous history of gestational diabetes mellitus: a 4-year prospective study. AB - OBJECTIVE: To investigate the potential use of the plasma homocysteine level as a predictor of diabetes in women with a previous history of gestational diabetes mellitus (GDM). RESEARCH DESIGN AND METHODS: At 6 weeks' postpartum, baseline examination was performed in 177 GAD-negative subjects. Of these subjects, 7 who were diagnosed with diabetes at baseline were excluded from further evaluation, and 170 with normal or impaired glucose tolerance (IGT) at baseline were followed annually over 4 years. The follow-up examinations included 2-h 75-g oral glucose tolerance tests (OGTTs), lipid profiles, homocysteine levels, anthropometric measurements, history taking, diet, and lifestyle. During the OGTTs, insulin and glucose levels were assayed every 30 min. Plasma homocysteine levels were determined by ion-exchange chromatography. RESULTS: Of the 170 women, 18 (10.6%) converted to diabetes during the 4-year follow-up period. Mean age, BMI, fasting insulin, and total cholesterol at baseline (6 weeks' postpartum test) were similar in the three study groups (i.e., normal, IGT, and diabetes). Fasting glucose levels, insulin-to-glucose ratios, and homocysteine levels were significantly higher in the diabetic group (P < 0.05). Higher glucose at the time of the diagnosis of GDM and higher homocysteine levels at baseline were independently associated with the onset of postpartum diabetes. These relationships were independent of age, BMI, and family history of diabetes. CONCLUSIONS: This prospective study identified homocysteine level as a significant risk factor for development of diabetes in women with previous GDM. PMID- 16249552 TI - Increased mortality risks of pre-diabetes (impaired fasting glucose) in Taiwan. AB - OBJECTIVE: The objective of this article was to assess mortality risks at different levels of fasting blood glucose (FBG) in Taiwan, with particular attention to those pre-diabetic subjects with impaired fasting glucose (IFG). RESEARCH DESIGN AND METHODS: Governmental employees and schoolteachers were followed up for an average of 11 years. With the use of Cox regression analyses, mortality risks were calculated for 36,386 subjects, aged 40-69. RESULTS: FBG > or =110 mg/dl was associated with increased mortality risks for all causes, cardiovascular diseases (CVD), and diabetes. IFG, when defined as 110-125 mg/dl, was associated with a significant increase for CVD and/or diabetes mortality. These mortality risks remained elevated when known CVD risk factors were adjusted for. The IFG group shared risk factor characteristics more with the FBG > or =126 mg/dl group than with the FBG <110 mg/dl group. When IFG was defined as 100-125 mg/dl, the number of subjects quadrupled, but mortality risks diminished substantially because of the inclusion of 100-109 mg/dl group. The lowest FBG group, 50-75 mg/dl, had a significant 2-fold risk from all causes. CONCLUSIONS: There was an overall J-shaped relationship between all-cause mortality and FBG. IFG, when defined as 110-125 mg/dl, is an independent risk factor and should be aggressively treated as a disease because its subsequent mortality risks for CVD and diabetes were significantly increased. The newly defined IFG at 100-125 mg/dl did not have the predictive power for later increases in CVD or diabetes mortality. PMID- 16249553 TI - High incidence of diabetes in men with sleep complaints or short sleep duration: a 12-year follow-up study of a middle-aged population. AB - OBJECTIVE: The aim of this study was to investigate the possible relationship among sleep complaints, sleep duration, and the development of diabetes prospectively over a 12-year period in a middle-aged Swedish population. RESEARCH DESIGN AND METHODS: A random sample of 2,663 subjects aged 45-65 years living in mid-Sweden were sent a postal questionnaire including questions about sleep complaints, sleep duration, sociodemographic characteristics, behavioral risk factors, medical conditions, and depression (response rate 70.2%). Twelve years later, a new questionnaire with almost identical questions was sent to all the survivors (n = 1,604) in 1995, and the questionnaire was answered by 1,244 subjects (77.6%). RESULTS: Men reporting new diabetes at follow-up more often reported short sleep duration (< or =5 h per night) (16.0 vs. 5.9%, P < 0.01), difficulties initiating sleep (16.0 vs. 3.1%, P < 0.001), and difficulties maintaining sleep (28.0 vs. 6.3%, P < 0.001) at baseline than men who did not develop diabetes. Women reporting new diabetes at follow-up reported long sleep duration (> or =9 h per night) more often at baseline than women not developing diabetes (7.9 vs. 2.4%, P < 0.05). In multiple logistic regression models, the relative risk (95% CI) for development of diabetes was higher in men with short sleep duration (2.8 [1.1-7.3]) or difficulties maintaining sleep (4.8 [1.9-12.5]) after adjustment for age and other relevant risk factors. Short or long sleep duration or sleep complaints did not influence the risk of new diabetes in women. CONCLUSIONS: Difficulties maintaining sleep or short sleep duration (< or =5 h) are associated with an increased incidence of diabetes in men. PMID- 16249554 TI - Racial/Ethnic differences in subclinical atherosclerosis among adults with diabetes: the multiethnic study of atherosclerosis. PMID- 16249555 TI - The impact of acute hypoglycemia on neuropsychological and neurometabolite profiles in children with type 1 diabetes. PMID- 16249556 TI - Renal malformations may be linked to mutations in the hepatocyte nuclear factor 1alpha (MODY3) gene. PMID- 16249557 TI - Population comparison of two clinical approaches to the metabolic syndrome: implications of the new International Diabetes Federation consensus definition. PMID- 16249558 TI - Efficacy of lifestyle education to prevent type 2 diabetes: a meta-analysis of randomized controlled trials. AB - OBJECTIVE: To evaluate the efficacy of lifestyle education for preventing type 2 diabetes in individuals at high risk by meta-analysis of randomized controlled trials, as assessed by incidence and a reduced level of plasma glucose 2 h after a 75-g oral glucose load (2-h plasma glucose). RESEARCH DESIGN AND METHODS: Through an electronic search, 123 studies were identified. A literature search identified eight studies that met strict inclusion criterion of meta-analysis for 2-h plasma glucose and five studies for the incidence of diabetes. All were randomized controlled trials of > or =6 months with lifestyle education that included a dietary intervention. Subjects were adults diagnosed as being at high risk for type 2 diabetes. The difference in mean reduction of 2-h plasma glucose from baseline to the 1-year follow-up and relative risk (RR) of the incidence of diabetes in the lifestyle education group versus the control group were assessed. Overall estimates were calculated using a random-effects model. Those estimates were confirmed by several models, and the possibility of selection bias was examined using a funnel plot. RESULTS: Lifestyle education intervention reduced 2 h plasma glucose by 0.84 mmol/l (95% CI 0.39-1.29) compared with the control group. The 1-year incidence of diabetes was reduced by approximately 50% (RR 0.55, 95% CI 0.44-0.69) compared with the control group. Results were stable and little changed if data were analyzed by subgroups or other statistical models. Funnel plots revealed no selection bias. CONCLUSIONS: Lifestyle education was effective for reducing both 2-h plasma glucose and RR in high-risk individuals and may be a useful tool in preventing diabetes. PMID- 16249559 TI - Ischemia imaging and plaque imaging in diabetes: complementary tools to improve cardiovascular risk management. AB - Cardiovascular disease is the most frequent cause of death and disability in diabetes, and the morbidity and mortality for coronary artery disease (CAD) in this population is two to four times higher than in nondiabetic subjects. Traditional risk factors do not fully explain the level of cardiovascular risk, and coronary disease events are often silent in diabetic patients. Thus, research has recently focused on improving the risk assessment of an individual patient with new tools in an effort to better identify subjects at highest risk and in need of aggressive management. Cardiovascular imaging has proven very helpful in this regard. Traditional methods to assess CAD are based on detection of obstructive luminal disease responsible for myocardial ischemia. However, acute coronary syndromes often occur in the absence of luminal stenoses. Hence, the utilization of imaging methodologies to visualize atherosclerosis in its presymptomatic stages has received mounting attention in recent years. In this article, we review the current literature on the utility of traditional imaging modalities for obstructive CAD (nuclear and echocardiographic stress testing) as well as atherosclerosis plaque imaging with carotid intima-media thickness and coronary artery calcium for risk stratification of diabetic patients. PMID- 16249560 TI - 2nd International Symposium on Triglycerides and HDL: lipid abnormalities and their treatment. PMID- 16249561 TI - Can protein kinase C beta-selective inhibitor, ruboxistaurin, stop vascular complications in diabetic patients? PMID- 16249563 TI - Glitazone in diabetes: relationship to patient dual public/private sector use. PMID- 16249562 TI - Postprandial hyperglycemia is a better predictor of the progression of diabetic retinopathy than HbA1c in Japanese type 2 diabetic patients. PMID- 16249564 TI - Observations on online services for diabetes management. PMID- 16249565 TI - Serum vitamin C levels in type 2 diabetic nephropathy. PMID- 16249566 TI - Should she or shouldn't she?: the relationship between infant feeding practices and type 1 diabetes in the genetically at risk. PMID- 16249567 TI - Pancreatic elastase-1 in stools, a marker of exocrine pancreas function, correlates with both residual beta-cell secretion and metabolic control in type 1 diabetic subjects: response to Cavalot et al. PMID- 16249568 TI - Pancreatic elastase-1 in stools, a marker of exocrine pancreas function, correlates with both residual beta-cell secretion and metabolic control in type 1 diabetic subjects: response to Mueller et al. PMID- 16249569 TI - Initiation of insulin therapy in patients with type 2 diabetes failing oral therapy: response to Mikhail and Cope and to Janka. PMID- 16249570 TI - No independent association of alanine aminotransferase with risk of future type 2 diabetes in the Hoorn study. PMID- 16249571 TI - Dying in the intensive care unit. PMID- 16249572 TI - Doctor of nursing practice: opportunity amidst chaos. PMID- 16249573 TI - Doctor of nursing practice: in need of definition. PMID- 16249574 TI - Letters to the editors. PMID- 16249575 TI - Letters to the editors. PMID- 16249576 TI - Letters to the editors. PMID- 16249577 TI - Letters to the editors. PMID- 16249578 TI - Letters to the editors. PMID- 16249579 TI - Letters to the editors. PMID- 16249580 TI - Letters to the editors. PMID- 16249581 TI - Letters to the editors. PMID- 16249582 TI - Letters to the editors. PMID- 16249583 TI - Letters to the editors. PMID- 16249584 TI - Relationship building and a story of hope in tsunami-ravaged Banda Aceh, Indonesia: perspective of a US Public Health Service nurse. PMID- 16249585 TI - Effect of frequency of manual turning on pneumonia. PMID- 16249586 TI - Critical care providers' perceptions of the use of vasopressin in cardiac arrest. AB - BACKGROUND: Although published algorithms and guidelines list epinephrine and vasopressin as either/or choices for treatment of ventricular fibrillation and/or pulseless ventricular tachycardia, little is known about how critical care providers respond to this recommendation. OBJECTIVES: To assess the use of vasopressin as a first-line drug of choice for ventricular fibrillation and/or pulseless ventricular tachycardia and describe factors that may influence decision making for using vasopressin. METHODS: A convenience sample from 4 academic medical centers in the United States was recruited to complete a 20-item survey on demographic factors such as year of last Advanced Cardiac Life Support (ACLS) provider course, specialty certification, predominant practice responsibility, and beliefs related to the use of vasopressin for cardiac arrest. Descriptive statistics, Pearson correlation analysis, and logistic regression were used to analyze the data. RESULTS: A total of 214 critical care providers (80% registered nurses) completed the survey. Year of last ACLS course (r=-0.188, P=.006) was a significant demographic factor, and behavioral beliefs (attitude about using vasopressin) had the strongest relationship (r=0.687, P<.001) and were the best predictor for intentions to use or recommend the use of vasopressin (beta=0.589, P<.001). CONCLUSIONS: Despite the recommendation for vasopressin as an agent equivalent to epinephrine for treatment of ventricular fibrillation and/or pulseless ventricular tachycardia, 63% of respondents used epinephrine as a first-line drug of choice. More research is needed to address the classification system for interpreting the quality of evidence that may influence practice. PMID- 16249587 TI - Family presence during resuscitation: a critical review of the literature. AB - Presence of patients' families during resuscitation has emerged as an important practice issue, sparking considerable controversy worldwide. Early advocates of allowing patients' families to be present during resuscitation faced more resistance than did current advocates because the former had little or no scientific research results to support their ideas. In the past 15 years, a number of quantitative studies, especially descriptive surveys, have been conducted. Qualitative researchers have also explored the lived experience of family members present during resuscitation and less commonly the perspectives of patients and health care providers. In this review of the literature, the current state of the science is critically reviewed and the ethical-theoretical perspectives of respective researchers and staff participants in the reviewed studies are discussed. Surveys were used to collect data in most studies to date. Limitations of these designs include small convenience samples, low response rates, use of retrospective surveys and the associated potential selection bias, and lack of consistency in survey instruments, factors that make comparison of findings between studies difficult. Recommendations to address the gaps in the current state of knowledge about family members' presence during resuscitation are discussed. Experimental and qualitative methods are especially needed to investigate the effect of family presence during resuscitation on patients, families, nurses and physicians, and other multidisciplinary staff members. PMID- 16249588 TI - Factors that enable or complicate end-of-life transitions in critical care. AB - BACKGROUND: Nurses in medical intensive care units are routinely involved in negotiations to maintain or withdraw life support. How nurses move from aggressively attempting to extend life to letting life end is not well understood. OBJECTIVE: To explore nurses' experiences of moving from cure- to comfort-oriented care and to describe factors that inhibit or facilitate such transitions. METHOD: A descriptive qualitative research design with brief observation of participants and focus group interviews was used. Participants were 19 female and 5 male nurses in an 18-bed medical intensive care unit in a 719-bed acute care hospital in the northeastern United States. RESULTS: The transition point between cure- and comfort-oriented care was unclear. Nurses reported that the patient's age, misunderstanding of the illness by the patient's family, family discord, and shifting medical care decisions made end-of-life transitions difficult. Conversely, developing a consensus among patients, patients' families, and staff about the direction of medical therapy; exhausting treatment options; and patients' lack of response to aggressive medical interventions helped nurses move toward comfort care. CONCLUSIONS: The most distressing situations for staff were dealing with younger patients with an acute life-threatening illness and performing futile care on elderly patients. End-of life transitions were difficult when patients' families had conflicts or were indecisive about terminating treatment and when physicians kept offering options that were unlikely to change patients' prognosis. The most important factor enabling nurses to move from cure- to comfort-oriented care was developing a consensus about the treatment. PMID- 16249589 TI - Moral distress of staff nurses in a medical intensive care unit. AB - BACKGROUND: Moral distress is caused by situations in which the ethically appropriate course of action is known but cannot be taken. Moral distress is thought to be a serious problem among nurses, particularly those who practice in critical care. It has been associated with job dissatisfaction and loss of nurses from the workplace and the profession. OBJECTIVES: To assess the level of moral distress of nurses in a medical intensive care unit, identify situations that result in high levels of moral distress, explore implications of moral distress, and evaluate associations among moral distress and individual characteristics of nurses. METHODS: A descriptive, questionnaire study was used. A total of 28 nurses working in a medical intensive care unit anonymously completed a 38-item moral distress scale and described implications of experiences of moral distress. RESULTS: Nurses reported a moderate level of moral distress overall. Highest levels of distress were associated with the provision of aggressive care to patients not expected to benefit from that care. Moral distress was significantly correlated with years of nursing experience. Nurses reported that moral distress adversely affected job satisfaction, retention, psychological and physical well being, self-image, and spirituality. Experience of moral distress also influenced attitudes toward advance directives and participation in blood donation and organ donation. CONCLUSIONS: Critical care nurses commonly encounter situations that are associated with high levels of moral distress. Experiences of moral distress have implications that extend well beyond job satisfaction and retention. Strategies to mitigate moral distress should be developed and tested. PMID- 16249590 TI - Development of the American association of critical-care nurses' sedation assessment scale for critically ill patients. AB - Clinicians commonly sedate critically ill patients. Sedatives should be administered to achieve predetermined end points. Most currently available scales used to assess sedation are inadequate because they focus on a single domain, such as consciousness. The development of the American Association of Critical Care Nurses' Sedation Assessment Scale is described. This new scale consists of 5 domains: consciousness, agitation, anxiety, sleep, and patient-ventilator synchrony. A major advantage of the scale is that its domains parallel common goals of sedation therapy for critically ill patients. The proposed measurements for each domain are based on a comprehensive evaluation of the science and expert recommendations. Before the scale is widely used, clinical testing is required to determine its validity and reliability in a variety of critically ill patients and care situations. PMID- 16249592 TI - Life-support technology, enframing, and disclosing. PMID- 16249591 TI - Novel spanish translators for acute care nurses and physicians: usefulness and effect on practitioners' stress. AB - BACKGROUND: Language barriers are significant impediments to providing quality health care, and increased stress levels among nurses and physicians are associated with these barriers. However, little evidence supports the usefulness of a translation tool specific to health care. OBJECTIVES: To evaluate the effectiveness of a novel English-Spanish translator designed specifically for nurses and physicians. The hypothesis was that the translator would be useful and that use of the translator would decrease stress levels among nurses and physicians caring for Spanish-speaking patients. METHODS: Novel English-Spanish translators were developed entirely on the basis of input from critical care nurses and physicians. After 7 months of use, users completed surveys. Usefulness of the translator and stress levels among users were reported. RESULTS: A total of 60% of nurses (n=32) and 71% (n=25) of physicians responded to the survey. A total of 96% of physicians and 97% of nurses considered the language barrier an impediment to delivering quality care. Nurses reported significantly more stress reduction than did physicians (P=.01). Most nurses and physicians had used the translator during the survey period. Overall, 91% of nurses and 72% of physicians found that the translator met their needs at the bedside some, most, or all of the time. All nurses thought that they most likely would use the translator in the future. CONCLUSIONS: The translator was useful for most critical care nurses and physicians surveyed. Health care providers, especially nurses, experienced decreased stress levels when they used the translator. PMID- 16249594 TI - Enhanced automaticity of the his bundle. PMID- 16249593 TI - The expanding role of the HMG-CoA reductase inhibitor, the most widely prescribed drug in the world. PMID- 16249595 TI - R. A. Fisher: an appreciation. PMID- 16249596 TI - Electromagnetic field exposure limitation and the future of MRI. PMID- 16249597 TI - Optical coherence tomography. AB - A review is presented of different scanning, acquisition and processing techniques used to obtain depth-resolved information in optical-coherence tomography (OCT). The principles and performances of different OCT techniques are discussed and images from different types of tissue are presented. Special attention is devoted to the progress in using the time-domain flying spot OCT technique and combination of the en face OCT imaging with confocal microscopy. Although OCT is based on white light interferometry, which is a well established and an old technology, the quest for higher resolution and faster acquisition of in vivo images has ensured OCT a rapid evolution in the last decade. Highly adventurous avenues to expand the OCT capabilities and trends are presented at the end of the review. PMID- 16249598 TI - Glioblastoma multiforme with atypical diffusion-weighted MR findings. AB - The aim of this study is to review the diffusion-weighted MRI findings of glioblastomas, to investigate those with atypical characteristics and to emphasise the reasons responsible for the atypical features on diffusion-weighted MR images. 48 cases of histologically proven glioblastomas were included in this study. In addition to conventional sequences of routine tumour protocol, diffusion-weighted MRI with spin-echo echo-planar sequence was performed. The cystic-necrotic components of the lesions, according to the conventional sequences, were determined on the diffusion-weighted MR images and were classified as typical or atypical. The presence of high signal intensity was accepted as an atypical feature while low signal intensity was accepted as typical. The apparent diffusion coefficient (ADC) values of the cystic components were calculated. The statistical significance of the typical and atypical glioblastomas was evaluated with the students t-test. In six of the cases apparent high signal intensity in diffusion weighted MR images was interpreted. In three cases the high signal intensity occupied all of the cystic component and in the other three most of the cystic component. The ADC values of the lesions varied between 0.86 x 10(-3) mm(2) s(-1) and 1.39 x 10(-3) mm(2) s(-1) (mean value 1.06+/-0.17 x 10(-3) mm(2) s(-1)). In 42 of the lesions the cystic-necrotic component demonstrated low signal intensity and the ADC values varied between 1.56 x 10(-3) mm(2) s(-1) and 3.32 x 10(-3) mm(2) s(-1) (mean value 2.36+/-0.46 x 10(-3) mm(2) s(-1)). The difference between ADC values of atypical and typical lesions was statistically significant (p<0.001). The vast majority of glioblastomas do not exhibit restricted diffusion in diffusion-weighted MRI, but some of them display homogeneous or heterogeneous high signal intensity and decrease of ADC values. Diffusion-weighted MRI alone is not helpful in the differentiation of malignant tumours from abscesses with low ADC values and similar conventional MRI findings. PMID- 16249599 TI - Influence of film digitization on radiological interpretation. AB - Our objective was to evaluate the influence of changing from analogue to digital imaging on interobserver and intraobserver image interpretation. Three radiologists interpreted 96 three image series of occipitomental radiographs of paranasal sinuses from the films and from the corresponding digitized images from the screen. Images were classified according to degree of abnormality as either normal, with mucosal thickening of less than 5 mm, with mucosal thickening of 5 mm or more, total opacity, air-fluid level or polyp or cyst of maxillary sinuses. In the present study we found that there were more differences between two radiologist's interpretations with a single method than in a single radiologist's interpretations between the methods, although radiologists interpreted fewer pathological findings from the digitized images than from the corresponding films. Our data show that the results of image interpretation are preferentially dependent on the reader rather than on the method of reading. PMID- 16249600 TI - Chasing "chasing the dragon" with MRI: leukoencephalopathy in drug abuse. AB - Spongiform leukoencephalopathy is a rare complication from inhalation of heated heroin vapour, a practice called "chasing the dragon". The MRI findings are considered pathognomonic, making MRI important for diagnosis. This is especially true in busy urban emergency departments where a variety of patients may present obtunded, unable or unwilling to provide a useful history. Even though the MR pattern of "chasing" toxicity is considered pathognomonic, there are mimickers. We compare the MRI findings of two classic cases of chasing leukoencephalopathy with one case of mimickery from cocaine exposure only. All three cases had diffuse symmetrical white matter changes. MR spectroscopy (MRS) in chasing patients showed increased lactic acid and myo-inositol, decreased N-acetyl aspartate and creatine, normal to slightly decreased choline, and normal lipid peak. MRS in the cocaine exposure patient showed marked increase in lactic acid and lipids. MR perfusion in one chasing patient was normal. IN CONCLUSION: (1) All three cases have MR findings suggestive of spongiform leukoencephalopathy. MRS may help differentiate toxicity due to inhaled heroin from other non-heroin related toxicities. (2) Discordance between perfusion and spectroscopy in one chasing patient adds evidence that the disease is due to impaired energy metabolism at the cellular level. (3) MR findings of spongiform leukoencephalopathy secondary to chasing heroin can progress despite apparent abstinence of the drug and during clinical improvement, suggesting the MR changes may represent an evolving injury. PMID- 16249601 TI - Coracoid tip position on frontal radiographs of the shoulder: a predictor of common shoulder pathologies? AB - The coracoacromial arch is a static anterior-superior stabilizer of the humeral head. Thus coracoacromial arch inclination, which varies depending on coracoid tip position, may be related to shoulder pathologies. Therefore, we retrospectively analysed coracoid tip positions in the true anterior posterior view of different shoulder pathologies: reference shoulders (n=27), shoulders with rotator cuff tear (supraspinatus tear n=29; subscapularis tear n=21) and shoulders with anterior glenohumeral instability (traumatic n=17; atraumatic n=14). In supraspinatus tear shoulders, the coracoid tip projected onto inferior glenoid half in 86% of cases (type I coracoid), extending more inferiorly compared with reference group (p=0.0002) or subscapularis tear shoulders (p<0.0001). In contrast, 78% of cases with subscapularis tear show the coracoid tip projected onto the superior glenoid half (type II coracoid). Atraumatic glenohumeral instabilities had a more superior coracoid tip position than traumatic instabilities (p=0.04), but no differences were observed on basis of coracoid type or in comparison with normal controls. We conclude that coracoid tip position is highly variable. Since type I coracoids are predominant in shoulders with supraspinatus tears and type II coracoids in shoulders with subscapularis tears, coracoid tip position may thus provide a simple diagnostic complement for a probable site of rotator cuff tears. PMID- 16249602 TI - Optical transillumination spectroscopy to quantify parenchymal tissue density: an indicator for breast cancer risk. AB - Mammographic screening for early detection of breast cancer has proven valuable in improving breast cancer survival. However, breast cancer incidence is still increasing, and thus preventative oncology needs to receive more attention, with the goal of identifying women with increased risk of developing breast cancer in the future and offering them risk reduction interventions. Mammogram derived parenchymal density pattern has been shown by various authors to provide a high odds ratio for breast cancer. Near-infrared optical transillumination spectroscopy was employed to determine physiological properties of the breast tissue to quantify differences in women with low or high breast cancer risk. Specifically in this study, women who had a recent mammogram underwent examination of their breast tissue by optical transillumination spectroscopy. Areas of adipose and glandular tissues which give rise to mammographic density patterns also have characteristic optical transillumination spectra. Correlation between optical transillumination spectroscopy and mammographic density pattern was established using partial least squares analysis. Results show that predicted tissue density based on optical transillumination spectroscopy correlates with mammographic observed tissue density, with a Spearman Rank correlation coefficient of 0.72. This suggests that optical transillumination spectroscopy may be a promising tool to quantify and monitor changes in breast cancer risk. PMID- 16249603 TI - Phase contrast X-ray imaging of mice and rabbit lungs: a comparative study. AB - The significant degree of X-ray phase contrast created by air-tissue interfaces, coupled with the poor radiographic contrast of conventional chest radiographs, makes the inflated lung an ideal candidate for investigating the potential diagnostic improvement afforded by phase contrast X-ray imaging. In small animals these methods highlight the lung airways and lobe boundaries and reveal the lung tissue as a speckled intensity pattern not seen in other soft tissues. We have compared analyser-based and propagation-based phase contrast imaging modalities, together with conventional radiographic imaging, to ascertain which technique shows the greatest image enhancement for various lung sizes. The conventional radiographic image of a mouse was obtained on a Siemens Nova 3000 mammography system, whilst phase contrast images of mice and rabbit chests were acquired at the medical imaging beamline (20B2) at the SPring-8 synchrotron radiation research facility in Japan. For mice aged 1 day, 1 week and 1 month old it was determined that analyser-based imaging showed the greatest overall image contrast, however, for an adult rabbit both techniques yielded excellent contrast. The success of these methods in creating high quality images for rabbit lungs raises the possibility of improving human lung imaging using phase contrast techniques. PMID- 16249604 TI - The risk of radiation-induced cancer in patients with squamous cell carcinoma of the head and neck and its results of treatment. AB - The purpose of this study was to determine the incidence and the results of treatment of cancer induced by radiotherapy for early stage (stage I and II) squamous cell carcinoma of the head and neck (SCH). The clinical records of 355 patients with early stage malignant lymphoma of the head and neck region treated by radiotherapy were reviewed, and then the records of 1358 patients with early stage SCH (oral cavity, 956; larynx, 154; oropharynx, 110; maxillary sinus, 86; lip, 20; epipharynx, 17; hypopharynx, 15) who underwent radiotherapy were reviewed. The disease-specific 10-year survival rate of the patients with 355 malignant lymphoma was 61%, and 5 cases of radiation-induced cancer occurred more than 8 years after irradiation. The crude incidence of radiation-induced cancer in the malignant lymphoma patients was 1.4%, and the 10-year probability by the actuarial life table method was 0.8%. The 10-year survival rate of the early stage SCH patients was 71%. The crude incidence of a second cancer in a previously irradiated field after an 8-year latent period (SCI) in the SCH patients was 1.8% (25/1358), and the 10-year probability was 1.6%. 12 SCIs were treated by surgery and 8 of those 12 patients (67%) resulted in success, whereas treatment by radiation resulted in failure in every other case. The risk of SCIs in the SCH group was higher than in the early stage malignant lymphoma group, although the difference was not statistically significant. The possibility of radiation-induced cancer in SCH is small, and the advantage of radiation therapy compares favourably with the risks of other treatments. PMID- 16249605 TI - Is carotid duplex scanning sufficient as the sole investigation prior to carotid endarterectomy? AB - Carotid endarterectomy (CEA) is the accepted treatment for certain patients who have had, or who are at risk of having, a stroke if they have a significant narrowing of the internal carotid artery. Rapid and accurate classification of the degree of stenosis is important as the benefit of surgery is highly dependent on this. The aim of this study was to assess whether the addition of angiography to duplex scanning resulted in a change in patient management in a unit where duplex scanning was used as the sole imaging investigation prior to CEA. The study population consisted of 64 patients with significant internal carotid artery stenosis on duplex scanning who were suitable for, and wished to be considered for, CEA. All patients underwent an angiogram. In this study 9 (14%) patients did not proceed to surgery on the basis of angiography and in a further 11 (17%) patients insufficient views of the distal vessel were obtained on duplex scanning. Three of these patients had extensive disease which excluded surgery. One patient experienced a transient ischaemic attack (TIA) at the time of angiography. In conclusion, this audit has highlighted the limitations in performing duplex scanning alone, and the costs that this can incur on the patient who may undergo an unnecessary operation. We cannot recommend duplex scanning as the sole investigation prior to CEA. There is need to evaluate the role of additional non-invasive carotid imaging such as magnetic resonance angiography or CT angiography in the assessment of these patients. PMID- 16249606 TI - Adrenal and renal metastases from follicular thyroid cancer. AB - Patients with differentiated thyroid cancer may have asymptomatic involvement of renal and/or adrenal gland, particularly if they are elderly and have associated metastases to other organs, which may remain undetected if these patients are not subjected to radioiodine treatment. Our experience also emphasises the role of routine post-radioiodine therapy whole body scan with high degree of clinical suspicion, which may reveal lesions otherwise not discernable in low dose whole body scan. All suspicious lesions should be subjected to structural imaging like ultrasound, CT or MRI for confirmation. In this setting, the role of radioiodine therapy is primarily aimed at palliation that might prolong their survival, probably reduce further spread and thus overall improve the quality of life. PMID- 16249607 TI - Pseudotumoural hemicerebellitis: imaging findings in two cases. AB - Pseudotumoural hemicerebellitis is exceedingly rare. It may closely resemble a tumour either clinically or on imaging, and a high index of suspicion is required to avoid unnecessary surgical procedures. MRI is the most useful diagnostic tool, and reveals a swollen cerebellar hemisphere, hyperintense in T(2) weighted images. A pattern of predominantly pial contrast enhancement, absence of a well defined mass and regression of the abnormalities in follow up examinations help to rule out malignancy. We report two cases of pseudotumoural cerebellitis (an 11 year-old girl and a 9-year-old boy) studied with CT and MRI. PMID- 16249608 TI - Colo-colonic intussusception caused by a solitary Peutz-Jeghers polyp. AB - A case of acute large bowel obstruction by colo-colonic intussusception in a healthy 19-year-old man is presented. The lead point of the intussusception is a rare solitary colonic Peutz-Jeghers polyp in the descending colon of a man without the full Peutz-Jeghers syndrome. The clinical presentation, imaging findings on plain radiographs, single contrast enema and CT, and findings at colonoscopy and surgery are correlated with pathology results and a brief review of the literature. PMID- 16249609 TI - Pseudocyst formation: a rare complication of wandering spleen. AB - Wandering spleen is a rare entity, in which the spleen is abnormally mobile due to its attachment by a long vascular pedicle. This long vascular pedicle predisposes it to various complications, the most common being torsion. Here, we present a case in which a wandering spleen in a young female was complicated by pseudocyst formation, and discuss the possible aetiology, pathogenesis, diagnosis and therapeutic implications of this extremely rare complication. PMID- 16249610 TI - Combined sphenoid and frontal sinus aplasia accompanied by bilateral maxillary and ethmoid sinus hypoplasia. AB - We describe CT scans of a case with bilateral aplasia of frontal and sphenoid sinuses with symmetrical hypoplasia of the ethmoid cellules and maxillary sinuses. This case appears to be first in the English-language literature with these combined findings. PMID- 16249611 TI - Low mortality of British radiologists. PMID- 16249612 TI - The emerging role of medical radiologists in new drug discovery. PMID- 16249613 TI - Paediatric medulloblastoma associated with poor prognosis and short volume doubling time. PMID- 16249614 TI - Linear no-threshold dogma. PMID- 16249615 TI - Factors for calculating the attenuation of barium plaster. PMID- 16249616 TI - Something old, something new? PMID- 16249617 TI - PFGE-based epidemiological study of an outbreak of Candida tropicalis candiduria: the importance of medical waste as a reservoir of nosocomial infection. AB - Between November 2002 and March 2003, an outbreak of candiduria occurred in the surgical intensive care unit (SICU) of a university-affiliated hospital in South Korea. This outbreak affected 34 patients and was caused by Candida tropicalis. To determine the source of the epidemic and the risk factors, surveillance cultures from the SICU, genotyping of Candida isolates by pulsed-field gel electrophoresis (PFGE), and a case-control study were performed. The surveillance cultures revealed that 6 environmental samples related to the urine disposal route were positive for C. tropicalis. The PFGE analysis of genomic DNA demonstrated identical band patterns for all of the C. tropicalis isolates obtained from SICU patients and the 6 environmental samples during the outbreak period, while epidemiologically unrelated strains showed unique PFGE band patterns. Although no risk factors were identified by the case-control study, this epidemiological investigation involving the use of molecular techniques suggests that improper disposal of infectious medical waste led to the cross transmission of a single clone that was responsible for the outbreak of C. tropicalis candiduria in this SICU. After implementing a better urine disposal system and thorough hand washing procedures, no further clusters of candiduria were detected in the SICU. PMID- 16249618 TI - Microsatellite polymorphism in the heme oxygenase-1 gene promoter is associated with susceptibility to cerebral malaria in Myanmar. AB - Cerebral malaria (CM) is a serious complication of Plasmodium falciparum malaria, and its pathogenesis leading to coma remains unknown. Heme oxygenase-1 (HO-1) catalyzes heme breakdown, eventually generating bilirubin, iron and carbon monoxide. The HO-1 gene promoter contains a polymorphic (GT)n repeat which may influence the expression level of HO-1. To explore the correlation between this (GT)n polymorphism and susceptibility to CM, we analyzed the frequencies of the (GT)n alleles in 120 Myanmarese patients with uncomplicated malaria (UM) and 30 patients with CM. The frequency of homozygotes for the short (GT)n alleles (<28 repeats) in CM patients was significantly higher than those in UM patients (P < 0.008, OR = 3.14). Thus, short (GT)n alleles represent a genetic risk factor for CM. PMID- 16249619 TI - Increased expression of the ska gene in emm49-genotyped Streptococcus pyogenes strains isolated from patients with severe invasive streptococcal infections. AB - Since 2000, emm49-genotyped Streptococcus pyogenes strains have been isolated from patients with severe invasive group A streptococcal infections in Japan, although they had not been isolated before 1999. We compare the characteristics of these strains with those of strains isolated from patients with non-invasive infections. SmaI-digested pulsed-field gel electrophoresis profiles of the isolates were almost indistinguishable between these groups, however, we found that ska (streptokinase gene) transcriptional levels in the strains isolated from patients with severe invasive infections were significantly higher than those in non-invasive infections. PMID- 16249620 TI - Is aerobic preputial flora age dependent? AB - Urinary tract infection (UTI) is one of the most commonly encountered infections in childhood. It has been demonstrated that the preputial sac can act as a reservoir of organisms and is thus responsible for causing ascending UTIs. This study was performed to determine the presence of preputial flora in different age groups. Prepuce and urine samples were taken simultaneously from 92 uncircumcised and healthy male children aged between 0-12 years. The data were analyzed by age, with 47 subjects of 6 years of age or less, and 45 aged 7-12 years. Twenty-seven percent of the older patients had negative preputial cultures versus 8% of those under 6 years of age (chi2 = 5.27, P = 0.02). In addition, enteric bacteria were the most common pathogens isolated from the prepuce in younger children while skin flora bacteria were most common in the older group (chi2 = 9.18, P = 0.002). The urine was sterile in all cases. Preputial cultures change with age in uncircumcised boys. This change may be related to the development of immune status, to histological or anatomical changes in the prepuce, and/or to improved personal hygiene. PMID- 16249621 TI - Prevalence of tetanus immunity in the Kocaeli Region, Turkey. AB - We assessed the antibody levels and risk factors for tetanus in an adult population in the Kocaeli Region of Turkey. In 595 individuals over 20 years of age, serum concentrations of anti-tetanus antibodies were detected by enzyme linked immunosorbent assay and a brief questionnaire was administered. The overall immunity rate was approximately 80% and there was a progressive decline in protection with increasing age. Antibody levels were significantly higher in educated people (80.1%), in employed people (90.3%), in those under 40 years of age (95.0%) and in those who stated that they had received a previous tetanus vaccination (87.9%). There was no association between antibody level and sex, residence, socioeconomic status, military status or history of injury. These data indicate that a vigorous post-injury prophylaxy with antitoxin and antisera should be put into practice, especially for older people, and that immunization programs applied every 10 years must be strengthened. PMID- 16249622 TI - Performance of strand displacement amplification assay in the detection of Chlamydia trachomatis and Neisseria gonorrhoeae. AB - This study is a critical analysis of certain amplification assays for detecting Chlamydia trachomatis and Neisseria gonorrhoeae infections which have demonstrated that the plasmid-free variant of C. trachomatis is frequently responsible for infection in our patients. Specifically, we evaluated the performance of the strand displacement amplification (SDA) assay in detecting either C. trachomatis or N. gonorrhoeae in 1,190 clinical samples, both urogenital and ocular, from 1,005 consecutive patients. The results obtained with the BDProbeTec ET System were compared with three referenced amplification methods for C. trachomatis (detecting the 16S rRNA gene, the omp1 gene and the plasmid of C. trachomatis) and with both the culture method as well as an amplification assay followed by genetic identification performed using the MicroSeq 500 16S ribosomal DNA-based system for N. gonorrhoeae. The sensitivity of SDA (76%) in detecting C. trachomatis is significantly low when compared with that of other molecular techniques employing 16S rDNA or omp1 as a target. The specificity of the methods for detecting C. trachomatis was excellent, ranging from 99.4 to 100%. Furthermore, the results of SDA in detecting N. gonorrhoeae also provided excellent results (100% specificity and sensitivity). PMID- 16249623 TI - Comparison of genomic structures in the serovar 1/2a Listeria monocytogenes isolated from meats and listeriosis patients in Japan. AB - Foodborne disease by Listeria monocytogenes, serovar 1/2a has recently been reported in many countries. Although contamination by this bacteria is also known to be gradually spreading among the marketed foods of Japan, there is little information on relation between listeriosis and food contamination. In the present study, the characteristics of the genomic structures of serovar 1/2a were compared among the isolates from marketed meats and listeriosis patients. Several isolates from meats purchased at the same shop on different days had the same genomic structure, and prolonged contamination was suggested by the conditions in the shop. Genomic structures of one strain isolated from meat were identical to those of two isolates from a patient. Another isolate was obtained from meats purchased at two different shops, and this isolate was also identical to that of the isolates from another patient. These findings suggest that the isolates from meat may have caused the listeriosis in the patients, and that the strains may have somehow traveled between the shops. PMID- 16249624 TI - Detection of Japanese encephalitis (JE) virus-specific IgM in cerebrospinal fluid and serum samples from JE patients. AB - Detection of Japanese encephalitis virus (JEV)-specific IgM by IgM-capture enzymed-linked immunosorbent assay (IgM-capture ELISA) has been accepted as the standard for serological diagnosis. In the present study, we analyzed the time course of the positive rate of JEV-specific IgM in serum and cerebrospinal fluid (CSF) specimens from confirmed JE patients. Serum and CSF samples were obtained from 155 JE cases for diagnostic purposes at hospitals in Thailand from 2002 to 2004. The levels of specific IgM were assessed by IgM-capture ELISA in the 171 serum and 156 CSF samples. Anti-JEV IgM was detected in 26 of 44 serum samples collected on days 1-4 of the disease period, in 31 of 44 samples collected on days 5-8, in 23 of 26 samples collected on days 9-12, and in all the samples collected on day 13 or later. Specific IgM was detected in 60 of 66 CSF samples collected on days 1-4 of illness, and in all the CSF samples but one collected on day 7 or later. The results indicate that the detection of JEV-specific IgM in CSF by IgM-capture ELISA is a reliable laboratory diagnostic method for confirmation of JE throughout the disease period, while the detection of IgM in serum samples is a reliable method on day 9 or later. PMID- 16249625 TI - Intensive care unit-acquired infections: incidence, risk factors and associated mortality in a Turkish university hospital. AB - In this prospective study, 93 intensive care unit (ICU)-acquired infections seen in 131 ICU patients were evaluated. Infection rates were found to be 70.9 in 100 patients and 56.2 in 1,000 patient-days. Pneumonia (35.4%) and bloodstream infections (18.2%) were the most common infections; Staphylococcus aureus (30.9%) and Acinetobacter spp. (26.8%) were the most frequently isolated microorganisms. The results of multivariate logistic regression analyses estimating the risk factors for ICU-acquired infections were as follows: length of stay in ICU (>7 days) (odds ratio [OR]: 7.02; 95% confidence interval [CI]: 2.80-17.56), respiratory failure as a primary cause of admission (OR: 3.7; 95% Cl: 1.41-9.70), sedative medication (OR: 3.34; 95% CI: 1.27-8.79) and operation (before or after admission to ICU) (OR: 2.56; 95% CI: 1.06-6.18). In logistic regression analyses, age (>60 years) (OR: 3.65; 95% CI: 1.48-9.0), APACHE II score >15 (OR: 4.67; 95% CI: 1.92-11.31), intubation (OR: 3.60; 95% CI: 1.05-12.39) and central venous catheterization (OR: 7.85; 95% CI: 1.61-38.32) were found to be significant risk factors for mortality. The difference in mortality rates between patients with ICU-acquired infection and uninfected patients was not statistically significant (mortality rates: 42.3 and 45.6%, respectively). A high incidence of nosocomial infections was found, and the risk factors for ICU-acquired infections and mortality were determined. PMID- 16249626 TI - Pathogen occurrence and antimicrobial susceptibility of urinary tract infection cases during a 20-year period (1983-2002) at a single institution in Japan. AB - Urinary tract infections are one of the most common infectious diseases diagnosed in outpatients as well as in hospitalized patients. Recently, urinary tract infections have become more complicated and difficult to treat. Therefore, the present study analyzed the occurrence and antimicrobial susceptibility of uropathogens isolated at Kobe University Hospital between 1983 and 2002. This study was performed with three patient groups: urology inpatients, urology outpatients, and inpatients of other departments. During the 20-year study period, we studied 15,925 urine isolates obtained from those patients who were diagnosed with urinary tract infection. Overall, Enterococcus faecalis was the most frequently isolated pathogen, followed by Pseudomonas aeruginosa and Escherichia coli. The frequency of Staphylococcus aureus increased over time, corresponding to an increase in the occurrence of methicillin-resistant S. aureus (MRSA). In addition, the rate of isolation of Serratia marcescens also increased over time, especially among patients with urinary tract catheters. Our results demonstrate that the uropathogens isolated at a single institution have shown a trend of increasing resistance to various classes of antimicrobial agents. In addition, serious problems should be anticipated in the treatment of multidrug resistant P. aeruginosa, fluoroquinolone-resistant E. coli, and arbekacin resistant MRSA. PMID- 16249627 TI - Molecular analysis of isoniazid, rifampin and streptomycin resistance in Mycobacterium tuberculosis isolates from patients with tuberculosis in Duzce, Turkey. AB - The aim of this study was to use DNA sequencing analysis to analyze the mutations in the most commonly targeted genes (katG, inhA, rpoB, rpsL) in isoniazid (INH)-, rifampin (RIF)- and streptomycin (SM)-resistant Mycobacterium tuberculosis strains obtained from subjects in Duzce, Turkey. Four isolates were found to be INH-resistant, 3 were RIF-resistant and 5 were SM-resistant, out of a total of 52 M. tuberculosis strains. In 3 of the 4 INH-resistant strains, a mutation in the katG gene in codon 315 appeared as AGC-->ACC (Ser-->Thr), and the other INH resistant strain showed a mutation in the katG gene in codon 314 as ACC-->CCC (Thr-->Pro). There were no mutations in the inhA gene in INH-resistant isolates. Two of the 3 RIF-resistant strains were found to have mutations in the rpoB gene in codon 516 appearing as GAC-->GTC (Asp-->Val), and the other RIF-resistant strain has a mutation in the rpoB gene in codon 531 as TCG-->TTG (Ser-->Leu). These 3 RIF-resistant strains are also INH-resistant. All 5 SM-resistant strains have mutations in the rpsL gene in codon 43 appearing as AAG-->AGG (Lys-->Arg). Thus, we found common gene mutations that bring about the resistance of M. tuberculosis to antituberculosis drugs in all of our isolates from Duzce. To the best of our knowledge, the ACC-->CCC (Thr-->Pro) mutation in the katG gene in codon 314 has not been previously defined. PMID- 16249629 TI - Detection of ehrlichial DNA in small rodents captured in a woodland area of Hokkaido, the northernmost island of Japan, where Lyme disease is endemic. AB - The ehrlichial gene was detected in small rodents trapped in a Lyme disease endemic area in Hokkaido, the northernmost island of Japan. Primer pairs of 16S rDNA targeting the genus Ehrlichia and other regions of the 16S rDNA specific for E. chaffeensis and E. muris were used for identification. The DNA fragment specific for 16S rDNA of Ehrlichia spp. was detected in 4 of 94 Apodemus speciosus mice (positive rate: 4.3%) and 5 of 73 Clethrionomys rufocanus bedfordiae mice (positive rate: 6.8%). The nucleotide sequence of the amplified 16S rDNA fragment was most similar to those of E. muris-like Ehrlichia, Ehrlichia spp. HF565 and Shizuoka-36, originating in the northern and central parts of Japan. In phylogenetic analysis based on 16S rDNA sequences, the northern, central and western groups of E. muris-like Ehrlichia from a cluster with microorganisms of the E. muris group. These results suggest that there are a group of E. muris microorganisms and a group of E. muris- like microorganisms in Japan. PMID- 16249628 TI - Genotoxic effect of ribavirin in patients with Crimean-Congo hemorrhagic fever. AB - In this study, we investigated the in vivo genotoxicity of ribavirin in humans, studying 3 patients with Crimean-Congo hemorrhagic fever who were treated with high-dose ribavirin. In order to evaluate genotoxicity, both the micronucleus (MN) test and the sister chromatid exchange (SCE) test were used. In all patients, blood samples were taken during and after therapy. Whole blood cultures were performed for 72 h and the MN assay and SCE test were then carried out to demonstrate the genotoxicity. In all patients, both SCE and MN amounts were found to be higher in the samples which were taken during therapy than in those at 1 month after therapy. The results of our study reveal that ribavirin has a reversible in vivo genotoxic effect on humans. PMID- 16249630 TI - Analysis of the molecular evolution of Listeria monocytogenes isolated from Japanese meats and environment. AB - Food contaminated by Listeria monocytogenes is a problem on a worldwide level because it is a serious food-borne pathogen. Although 3 evolutionary divisions have been reported for L. monocytogenes, the evolution of Japanese isolates has not yet been clarified. Thus, in order to determine the lineage of these Japanese isolates, we classified and conducted phylogenetic analysis of 407 bp (position 1116-1522) of the iap gene derived from 88 isolates from Japanese listeriosis patients, foods and environment. The isolates were classified into 18 types commonly accompanied by serotypes, and the types were divided into 3 lineages. Our results suggest that these Japanese isolates belong to the 3 lineages of L. monocytogenes isolated in other countries. PMID- 16249631 TI - Nosocomial methicillin-resistant Staphylococcus aureus endocarditis with splenic abscess in a pregnant woman. AB - A 36-year-old, 7-week-gravida patient with catheter-related nosocomial infective endocarditis due to methicillin-resistant Staphylococcus aureus (MRSA) is presented in this paper. The patient was admitted to our hospital because of carbon monoxide intoxication. After 14 days, MRSA catheter-related bacteremia developed. The central venous catheter was immediately removed, and teicoplanin therapy was started. Because of persistent fever, leukocytosis, and high C reactive protein values, endocarditis was suspected. A transesophageal echocardiogram revealed 19-mm vegetation on her mitral valve, confirming the diagnosis of endocarditis. Gentamicin and rifampicin were added to the therapy regimen, and the dose of teicoplanin was increased to 12 mg/kg-day. After 8 days, a splenic abscess was detected by ultrasonography. Vegetation excision, mitral valve replacement by open-heart surgery and splenectomy were performed in the same operation. Antibiotherapy was continued for 6 weeks after surgery, and the patient's condition improved. The development of endocarditis could be prevented by proper clinical practices. PMID- 16249632 TI - Comparison of the incidence of norovirus infection in homes for the elderly. PMID- 16249633 TI - A coprological survey of the potential definitive hosts of Echinococcus multilocularis in Aomori Prefecture. PMID- 16249634 TI - An outbreak of mixed infection of enterohemorrhagic Escherichia coli O26:H11 and norovirus genogroup II at a kindergarten in Shimane, Japan. PMID- 16249635 TI - Epidemic of hand, foot and mouth disease in Kawasaki City, Japan. PMID- 16249636 TI - Cryptosporidium outbreak in a sports center. PMID- 16249637 TI - Outbreak of enterohemorrhagic Escherichia coli O111 among high school participants in excursion to Korea. PMID- 16249639 TI - Telomerase activity--a prognostic factor in colorectal cancer? PMID- 16249638 TI - Trastuzumab and breast cancer. Are we just beyond the prologue of a fascinating story? PMID- 16249640 TI - Telomerase activity as a prognostic factor in colorectal cancer. AB - BACKGROUND: As the enzyme telomerase extends the life of the cell through its ability to lengthen telomeres, its activity in different types of tumor has been evaluated as a possible factor involved in tumorigenesis. The aim of this study was to assess the prognostic significance of telomerase activity in patients with colorectal carcinoma. PATIENTS AND METHODS: Telomerase activity was determined in 103 patients undergoing surgery for colorectal cancer between 2001 and 2003. Telomerase activity was determined by an enzyme-linked immunoassay based on the amplification of telomeric repeat sequences (TRAP assay). RESULTS: 90% of our study population showed telomerase activity. Telomerase activity was related to tumor stage and site: a lower proportion of patients with stage A tumors showed telomerase activity compared to more advanced stages; and more patients with colon than with rectal carcinomas were telomerase positive. Multivariate analysis revealed that by adjusting for tumor stage, telomerase activity could be used to predict the risk of death or recurrence (p < 0.001). CONCLUSIONS: Activation of telomerase seems to be a frequent event related to the stage of the tumor in colorectal tumorigenesis. Our findings suggest that telomerase activity can predict a greater risk of death or recurrence, irrespective of the more conventional prognostic factors. PMID- 16249641 TI - Safety and efficacy of trastuzumab every 3 weeks combined with cytotoxic chemotherapy in patients with HER2-positive recurrent breast cancer: findings from a case series. AB - BACKGROUND: Trastuzumab has been repeatedly shown to result in significant clinical benefits and was subsequently accepted as the treatment of choice for HER2-positive advanced breast cancer - particularly as first-line treatment in combination with taxanes and as monotherapy in the second-line or third-line setting. Trastuzumab is currently licensed as a weekly treatment, although a 3 weekly schedule could be used conveniently in combination with other cytotoxic agents that are administered on a 3-weekly basis in metastatic breast cancer. PATIENTS AND METHODS: We determined the safety of i.v. trastuzumab (8 mg/kg followed by 6 mg/kg) every 3 weeks in combination with chemotherapeutic agents administered in 3-weekly courses (docetaxel, vinorelbine and capecitabine) in 31 patients with HER2-positive recurrent locoregional and/or metastatic breast cancer. RESULTS: 3-weekly trastuzumab appeared to be as well tolerated as the standard once-weekly schedule. All myelosuppressive adverse events and the majority of non-hematological adverse events were typical and characteristic of the individual concomitant cytotoxic agents. Transient trastuzumab-related infusion reactions occurred in 5 patients and 1 patient developed cardiac dysfunction, which recovered after discontinuation of trastuzumab. Efficacy appeared favourable: 18 clinical responses (3 complete and 15 partial) and 8 disease stabilizations gave an overall response rate of 58% (70% in the 20 patients receiving first-line therapy). Median progression-free and overall survival times were 9.9 months (95% CI: 6.3-13.5) and 23.1 months (95% CI: 19.2 27.0), respectively. CONCLUSIONS: These findings will likely encourage further evaluation of this more convenient 3-weekly trastuzumab regimen in patients with HER2-positive metastatic breast cancer. PMID- 16249642 TI - Gemcitabine in the treatment of relapsed and refractory Hodgkin's disease. AB - BACKGROUND: Patients with refractory Hodgkin's disease or relapsing after high dose therapy and autografting have a poor prognosis. Here, we present our experiences with gemcitabine in this setting. PATIENTS AND METHODS: We treated 14 patients with relapsed or refractory Hodgkin's disease with gemcitabine. The treatment was given on a compassionate use basis, off-label and not according to a study protocol. Patients were 17-46 years of age. 1 patient had stage IA disease, 2 patients had stage IIIB disease and 11 patients had stage IVB disease. 9 patients had received radiotherapy. 8 patients had been autografted and 1 patient auto- and allografted. Gemcitabine was administered at a starting dose of 1 g/m(2) on days 1 and 8 every 3 weeks in combination with steroids. RESULTS: The median follow-up period was 10 months. Hematological toxicity grade 3-4 occurred in 12 patients leading to dose reductions. 1 patient died of neutropenic sepsis. No other non-hematological toxicities were observed. The response rate was 64% with 6 patients achieving complete remission (CR) and 3 patients partial remission (PR). The median time to treatment failure was 9 months, and survival was 11 months. Responses were seen in previously transplanted patients and in patients refractory to previous treatment. The so far longest responder has been in CR for over 68 months. CONCLUSION: Gemcitabine is an effective treatment for Hodgkin's disease. Heavily pretreated patients often require dose reductions. PMID- 16249643 TI - Characterization of prognostic factors and efficacy in a phase-II study with docetaxel and estramustine for advanced hormone refractory prostate cancer. AB - BACKGROUND: Docetaxel based chemotherapy not only reduces pain and improves quality of life in advanced hormone refractory prostate cancer (HRPC), but it also improves survival. We investigated the combination of docetaxel and estramustine in patients with HRPC regarding efficacy and prognostic parameters. PATIENTS AND METHODS: We conducted a phase-II trial, administering docetaxel (70 mg/m(2) i.v., day 2, every 3 weeks) and estramustine (280 mg 3 times daily p.o., 1 day prior to docetaxel, on 5 consecutive days) to patients with HRPC. Patients were monitored for PSA (prostate-specific antigen) response and toxicity. RESULTS: 62 patients were treated. The median age was 67.5 years, the median PSA was 177.9 ng/ml. The median number of cycles was 6. The median time to progression (TTP) and median survival time were 14 (+/-2) and 24 (+/-5) months, respectively. A = 50% decrease in PSA levels from baseline occurred in 38 (61.3%) patients of whom 25 (40.3%) had a = 75% PSA decrease. The main grade 3-4 hematologic toxicities were neutropenia 34% and anemia 18%. CONCLUSIONS: The combination of docetaxel and estramustine exerts substantial activity in HRPC suggesting an overall survival benefit with manageable toxicity. This trial also demonstrated a survival advantage for patients with early chemotherapeutic intervention. We identified PSA relapse, baseline PSA and hemoglobin as valuable prognostic factors in this setting. PMID- 16249644 TI - Prolonged survival of patients receiving trastuzumab beyond disease progression for HER2 overexpressing metastatic breast cancer (MBC). AB - BACKGROUND: The aim of this retrospective analysis was to evaluate the impact of trastuzumab-based regimens on the survival of patients with HER2-overexpressing metastatic breast cancer (MBC). The study specifically focussed on the influence of the continuation of trastuzumab-based treatment despite tumor progression on survival. PATIENTS AND METHODS: Patients with HER2 overexpressing MBC were included in this retrospective analysis. HER2 overexpression was determined by the immunohistochemical staining score (DAKO Hercep Test). Trastuzumab was applied at a loading dose of 4 mg/kg and a maintenance dose of 2 mg/kg. RESULTS: Among 136 HER2 overexpressing patients (DAKO score 3+), 66 patients received first-line trastuzumab, 47 patients received trastuzumab as second-line therapy and 23 patients received trastuzumab beyond disease progression. There was no significant difference regarding the duration of trastuzumab-based treatment (first-line: 29.5 weeks vs. second-line: 25 weeks). Moreover, there was no difference in the response rate (first-line: 37.9% vs. second-line: 35.7%) or the median survival (p = 0.47 log rank). Patients who received = 2 trastuzumab-based regimens for MBC survived significantly longer compared to those who had received only 1 regimen (= 2 regimens: 62.4 months vs. 1 regimen: 38.5 months; p = 0.01 log rank). CONCLUSIONS: Trastuzumab is highly effective in the treatment of HER2 overexpressing MBC. Compared to historical controls, overall survival appears to be markedly prolonged, particularly in patients who received sequential trastuzumab-based treatment beyond disease progression. PMID- 16249645 TI - Pharmacogenetics of extraordinary responses to 5-FU/cisplatin chemotherapy in advanced gastric cancer -- report of 2 cases. AB - BACKGROUND: Gastric cancer is often diagnosed in the metastatic stage, and only 10% of patients survive for as long as 2 years. Current chemotherapy regimens show significant treatment-related toxicities. It is crucial to identify the patients that will benefit most from certain chemotherapy regimens in order to avoid unnecessary side effects. PATIENTS AND METHODS: 2 patients with advanced gastric cancer repeatedly received 5-FU/cisplatin combination chemotherapy. Genomic DNA was extracted from tumor tissue and mononuclear blood cells. Genotype analysis of genes of metabolizing and DNA repair enzymes was carried out using a PCR-RFLP technique. Direct sequencing was used to identify mutations of the gene dihydropyrimidine dehydrogenase (DPD). RESULTS: Prolonged survival of 51 and 29 months, respectively were observed in our 2 patients. Both patients were positive for genotypes of thymidylate synthase -- the target enzyme of 5-FU -- that are associated with improved drug response. DPD variants connected with increased toxicity were not observed. However, both patients also showed genotypes in cisplatin metabolizing enzymes which enhance the effect of the drug. CONCLUSION: Genotype analysis in drug metabolizing enzymes of 5-FU and cisplatin provide a possible explanation for extraordinary therapy effects observed in 2 patients with advanced gastric cancer. PMID- 16249646 TI - Pegfilgrastim -- rational drug design for the management of chemotherapy-induced neutropenia. AB - Neutropenia is the most important dose-limiting toxicity of myelotoxic chemotherapy. Current guidelines recommend primary prophylactic use of granulocyte colony stimulating factor (G-CSF) with chemotherapy regimens associated with an incidence of febrile neutropenia (FN) of at least 40% and in patients at high risk of infections, such as the elderly. Using prophylactic G CSF support, planned chemotherapy doses are administered on time more frequently. Pegfilgrastim is a rationally designed recombinant human G-CSF with a sustained duration of action. A once-per-cycle 6-mg fixed dose of pegfilgrastim reduced the duration of severe neutropenia and the incidence of FN as efficiently as daily filgrastim in standard or dose-dense chemotherapy regimens in young and elderly patients with breast cancer, non-small-cell lung cancer and lymphomas. The safety profile of onceper- cycle pegfilgrastim is comparable with that of daily filgrastim. In conclusion, a fixed-dose of pegfilgrastim given once per cycle is a suitable substitute for body weight-based daily dosing of G-CSF, an improvement which should be particularly beneficial for outpatients receiving myelotoxic chemotherapy. PMID- 16249647 TI - AIDS research in Brazil. PMID- 16249648 TI - Self-perception of body changes in persons living with HIV/AIDS: prevalence and associated factors. AB - BACKGROUND: Highly active antiretroviral therapy has brought about a substantial improvement in the prognosis of HIV/AIDS. In this context, therapy-related body changes (lipodystrophy) gain in importance, in light of the psychological distress they cause and of their association with adherence to treatment. This study analyses patients' self-perception of central fat gain (CFG) and peripheral fat loss (PFL). METHODS: A total of 457 patients were interviewed in a university outpatient facility for the treatment of adults and adolescents with HIV/AIDS in the city of Sao Paulo, Brazil, between September and December 2001. RESULTS: Two thirds of subjects (64.3%) perceived body changes. The self-perception of CFG and PFL was associated with greater schooling. The self-perception of CFG was more frequent among women and in patients who used protease inhibitors for longer periods. The self-perception of PFL was more frequent among older patients, patients who used stavudine for longer periods, and patients who reported a lack of adherence to antiretroviral agents. The quality of affective/social relationships with friends and family was inversely associated with the self perception of PFL. CONCLUSION: The evaluation of self-perceived body changes and their determinants in individuals living with HIV/AIDS may help improve provided care. Listening to what patients have to say concerning antiretroviral therapy related body changes and how they perceive them, as well as including the patient in therapeutic decisions in this regard will contribute towards greater adherence to proposed interventions and towards an improvement in the quality of life. PMID- 16249649 TI - Survival of AIDS patients using two case definitions, Rio de Janeiro, Brazil, 1986-2003. AB - BACKGROUND: Recent studies have shown substantial increases in the survival of AIDS patients in developed countries and in Brazil as a result of antiretroviral therapy (ART) and prophylaxis for opportunistic infections. This study compares survival rates using the Brazilian Ministry of Health 2004 and Centers for Disease Control and Prevention (CDC) 1993 case definitions in a large HIV/AIDS referral centre in Rio de Janeiro. METHODS: Survival after AIDS diagnosis was assessed in a clinic-based cohort of 1415 individuals using the Kaplan-Meier method and Cox proportional hazards models. RESULTS: There were 393 (88%) deaths from AIDS-related causes and 52 (12%) from unrelated or unknown causes. A total of 205 patients (14%) were lost to follow-up and 765 patients (55%) remained alive until the end of the study. Three-quarters of patients (75%) were still alive 22 months [95% confidence interval (CI) 19-26] after the AIDS diagnosis according to the CDC case definition and 31 months (95% CI 26-36) according to the Ministry of Health case definition. Independent predictors of survival included AIDS defined by CD4 cell count and any use of highly active antiretroviral therapy, with either case definition, and initial stage of the case, with the Ministry of Health case definition. CONCLUSION: Survival observed in this reference centre is comparable or longer than other international studies, although the choice of case definition criterion influenced findings. Adoption of the Ministry of Health case definition may enhance the ability to track the use of and outcomes from ART among AIDS patients. PMID- 16249650 TI - Characteristics and survival of AIDS patients with hepatitis C: the Brazilian National Cohort of 1995-1996. AB - BACKGROUND: As AIDS patients live longer, the management of co-morbidities becomes increasingly important. Previous studies from developed countries give conflicting results as to whether co-infection with hepatitis C virus (HCV) lowers the life expectancy of individuals with AIDS. METHODS: This retrospective cohort study was based on a medical record review of a nationally representative sample of 2821 adult AIDS cases diagnosed in 1995 and 1996 in Brazil. We compared the characteristics and survival of patients known to be positive and negative for HCV. RESULTS: A total of 833 patients received HCV testing, and the prevalence was 33%. HCV-positive patients received less intensive antiretroviral treatment. The crude mortality was greater for HCV-positive patients (hazard ratio 1.26; P = 0.04), but HCV status was not a significant predictor in a multivariate analysis that included other predictors of survival. CONCLUSION: Brazilian AIDS patients with hepatitis C have a shorter survival than those without, but this seems to be mainly as a result of their receiving less antiretroviral treatment. We cannot say whether this is because of the fear of hepatotoxicity, an inability to tolerate treatment, or for other reasons. To improve survival, these patients need optimal treatment of their HIV disease. PMID- 16249651 TI - Optimistic perception of HIV/AIDS, unprotected sex and implications for prevention among men who have sex with men, Sao Paulo, Brazil. AB - BACKGROUND: This study examines the association between optimistic perceptions about AIDS and unprotected sex among men who have sex with men (MSM) in the city of Sao Paulo, Brazil. METHODS: A cross-sectional study was carried out among MSM in leisure areas of Sao Paulo in 2003. We interviewed 161 participants aged 18-30 years. RESULTS: Thirty-nine per cent (95% confidence interval 32-47%) reported unprotected anal sex with steady or casual partners in the previous 6 months. The optimistic perception score created for this study was associated with unprotected sex (P = 0.01) and higher education (P = 0.02). The quartile with the most optimistic perception was 1.8 times more likely to engage in unprotected anal sex compared with the quartile with the least optimistic perception. CONCLUSION: This study suggests that the current situation regarding AIDS, which is seemingly favourable, may create optimistic perceptions leading to unprotected sexual practices. Prevention programmes, particularly for MSM, need to take this into account. PMID- 16249652 TI - Prevention of mother-to-child transmission of HIV in Sao Paulo State, Brazil: an update. AB - BACKGROUND: Sao Paulo State has had the largest number of paediatric AIDS cases in Brazil. Since 1996, Sao Paulo (and Brazil nationally) has implemented an aggressive programme to reduce perinatal transmission. We have gathered available indicators to examine the programme's impact. METHODS: We obtained data on reported AIDS cases from the AIDS surveillance system; data on the number of mother/infant pairs treated with zidovudine from the state logistics office responsible for distributing HIV medication; and the rates of perinatal transmission from a multicity study of the Brazilian Pediatric Society that includes a number of Sao Paulo facilities, which were compared with an independent study in 1995. The years for which data were available varied according to the source of the indicator. RESULTS: Annual reported cases of AIDS as a result of perinatal transmission fell 58.9% from 1997 to 2002. The number of cases treated with zidovudine increased 73.7% from 1997 to 2004. The rate of perinatal transmission among babies born to HIV-positive mothers fell from 16% in 1995 to 2.4% in 2002 in the reference clinics that participated in the Brazilian Pediatric Society study. CONCLUSION: Both process and outcome indicators point to the effectiveness of efforts to reduce perinatal transmission in Sao Paulo State. PMID- 16249653 TI - Factors associated with condom use among youth aged 15-24 years in Brazil in 2003. AB - OBJECTIVE: To analyse factors associated with the lack of condom use among young people at last sexual intercourse with a steady or casual partner. DESIGN: A cross-sectional study involving 1170 household interviews and designed to build a representative sample of the population of young Brazilian residents aged 15-24 years (2003). METHODS: In the multivariate analysis of data, non-conditional logistic regression modelling was applied to assess the determinants of condom use at last sexual intercourse among young people with steady or casual partners. RESULTS: The overall level of condom use at last sexual intercourse was high (60%), although it was significantly more common in casual sexual partnership. Cohabitation was associated with a lack of condom use in both casual and steady partner encounters. In addition, being female, having less schooling, having no work history, and per capita family income above the minimum wage were factors related to not using condoms in the group of young people who had their last sexual encounter with steady partners. Among young people with casual partners, such factors included a positive history of alcohol use, first sex at 9-16 years of age, inadequate knowledge of AIDS treatability and bereavement related to violence. CONCLUSION: This study confirmed that the determinants of condom use among youth during last sexual intercourse vary according to whether the partner was casual or steady. Prevention campaigns should develop specific messages for each of these contexts. PMID- 16249655 TI - Knowledge, practices and behaviours related to HIV transmission among the Brazilian population in the 15-54 years age group, 2004. AB - OBJECTIVE: To describe transmission vulnerability for acquiring HIV infection among the Brazilian population aged 15-54 years. DESIGN: A population-based survey. METHODS: Sampling was stratified by geographical region. A total of 6006 interviews were conducted. Indicators of knowledge and sexual practices and the relative sizes of the vulnerable subgroups were estimated. Logistic regression analysis was used to determine the main factors associated with safe sex practices. RESULTS: Regarding knowledge indicators in the age group 15-24 years, a high percentage (91%) spontaneously cited sexual intercourse as a form of HIV transmission, and 62% had correct knowledge of the modes of HIV transmission (five correct items). The proportion of consistent condom use with casual partners was 52%, increasing to 59% in the youngest age group. Higher proportions of inconsistent condom use with any kind of partner were found among women and among the poorest. A multiplicity of sexual partners, low socio-economic status and cocaine use were important predictors of unprotected sex among men living without a companion. Among individuals aged 15-49 years, 0.2% currently inject cocaine, 4.6% of the men paid for sex at least once over the past year and 1.0% of the women were paid in exchange for sex. Among sexually active men of the same age group, 3.5% reported sexual relations with other men. CONCLUSION: Besides the need to establish the role exercised by the vulnerable subgroups in the HIV transmission dynamics, results indicate that it is necessary to investigate unsafe sexual practices further among the poorer sectors of society. PMID- 16249654 TI - Non-adherence among patients initiating antiretroviral therapy: a challenge for health professionals in Brazil. AB - OBJECTIVE: To assess the incidence, magnitude and factors associated with the first episode of non-adherence for 12 months after the first antiretroviral prescription. DESIGN: A prospective study of HIV-infected patients receiving their first antiretroviral prescription in public referral centers, Belo Horizonte, Brazil. Baseline assessment occurred at the moment of the first prescription and follow-up visits at the first, fourth and seventh month, from May 2001 to May 2003. METHODS: Non-adherence was self-reported and defined as the intake of less than 95% of the prescribed doses for 3 days before the follow-up interviews. Cumulative and person-time incidence were estimated and Cox's proportional model was used to assess the relative hazard (RH) of non-adherence with 95% confidence interval for both univariate and multivariate analysis. RESULTS: Among 306 patients, the cumulative incidence of non-adherence was 36.9% (incidence rate 0.21/100 person-days). Multivariate analysis (P < 0.05) showed that unemployment (RH = 2.17), alcohol use (RH = 2.27), self-report of three or more adverse reactions (RH = 1.64), number of pills per day (RH = 2.04), switch in antiretroviral regimen (RH = 2.72), and a longer time between the HIV test result and the first antiretroviral prescription (RH = 2.27) were associated with an increased risk of non-adherence, whereas the use of more than one health service indicated a negative association (RH = 0.54). CONCLUSION: The current analysis has pointed out the importance of clinical and health service characteristics as potential indicators of non-adherence after initiating therapy. Early assessment and intervention strategies should be priorities in these AIDS public referral centres. Feasible and reliable indicators for the routine monitoring of adherence should be incorporated in clinical practice. PMID- 16249656 TI - Factors associated with institutionalization of children orphaned by AIDS in a population-based survey in Porto Alegre, Brazil. AB - BACKGROUND: There are increasing numbers of children orphaned by AIDS, especially in countries without universal free AIDS treatment. As institutionalization is associated with bad health and developmental outcomes, we have identified the factors associated with the institutionalization of AIDS orphans in a population based survey in a city in southern Brazil. METHODS: Using AIDS mortality and healthcare registries from 1998 to 2001, a cross-sectional study was conducted among the caregivers of children aged 0-14 years who were the survivors of parents dying of AIDS in Porto Alegre. Data were collected by a household survey using a structured questionnaire. RESULTS: Out of 1131 orphans identified, 75.4% of their caregivers participated. Among participants, 70% had lost their father and 50% their mother, and 21% had lost both parents. At the time of the survey, 41% of the children lived with the mother, 25% lived with grandparents and 5% lived in institutions. In multivariate analysis, HIV positivity multiplied the child's chances of living in an institution by a factor of 4.6, losing its mother by 5.9, losing both parents by 3.7, and having a non-white mother by 4.0. CONCLUSION: This study provides population-based data on what has become of the children of individuals dying of AIDS. Improving the quality of life and averting the institutionalization of AIDS orphans requires interventions to promote the survival of mothers living with AIDS, as well as specific interventions for child family placement. Reducing the stigma of HIV infection in children and racial discrimination present challenges in Brazil. PMID- 16249657 TI - Sexually transmitted disease/HIV risk behaviour among women who have sex with women. AB - OBJECTIVE: To analyse the epidemiological aspects of sexually transmitted diseases (STD) among women who have sex with women (WSW) in Sao Paulo, Brazil. METHOD: A cross-sectional study with interviews and analysis of clinical and gynaecological tests in women, by means of a convenience sample. Characteristics were gathered according to age, sociobehavioural profile, reproductive life and sexuality. RESULTS: The study included 145 women. They started sexual activity at an average age of 16.9 years, and 23.4% of them had had heterosexual relations during the preceding year, with a relatively low frequency of condom use. In sexual relations with women, 54.5% used condoms when they shared sex toys. A previous STD was reported by 38% of them. The following STD were diagnosed: trichomonas (3.8%), bacterial vaginosis (33.8%), fungi (25.6%), Chlamydia (1.8%), hepatitis B (7%), hepatitis C (2.1%), abnormal Pap smear (7.7%), human papillomavirus (6.2%) and HIV (2.9%). CONCLUSION: In this study, many WSW did not report a single risk behaviour, but often reported a combination of several potential risk factors. Therefore, one cannot speak of high or low-risk behaviour for STD/HIV, but rather of multiple-risk behaviour. It is evident that there is a need for healthcare professionals to be correctly informed and sensitive towards the healthcare of WSW. PMID- 16249658 TI - Evaluation of rapid tests for anti-HIV detection in Brazil. AB - OBJECTIVES: This assessment in Brazil was to evaluate the performance of commercially available HIV rapid test (RT) against the gold standard testing and to establish a highly sensitive and specific RT algorithm for HIV diagnosis. DESIGN: A prospective, anonymous and unlinked study. METHODS: An evaluation of seven commercially available RT to compare their performance against the gold standard tests for Brazil. This includes two competing enzyme immunoassays plus a Western blot for confirmation. After informed consent, whole blood samples were collected from volunteers in voluntary counselling and testing sites (n = 400), antenatal clinics (n = 500) and from HIV-positive controls in AIDS treatment centres (n = 200). Two seroconversion panels, one HIV-1 subtype (B, B', C and F) panel and an operational assay performance evaluation were also part of the study parameters. RESULTS: For the seven RT the clinical sensitivity ranged from 97.74 to 100% and clinical specificity from 99.43 to 100%. However, only four RT were considered acceptable after full evaluation. The two EIA had a clinical sensitivity of 100% and clinical specificity of 99.32 and 99.66%. Two RT had the same performance on the seroconversions panels as the EIA. The operational assay performance evaluation for the RT indicated that Hexagon and Capillus could not be classified as simple assays. CONCLUSION: We have provided evidence that RT assays can perform equally or better than EIA for the detection of HIV antibodies. The simplicity and rapidity of the RT warrants its utilization in an algorithm for a rapid diagnosis of HIV infection. PMID- 16249659 TI - Estimating the genetic component (RGC) in pharmacokinetic variability of the antiretroviral didanosine among healthy Brazilians. AB - BACKGROUND: We estimate the variance between between- and within-subjects, using mixed effect models, as a way to assess the genetic component in explaining the observed heterogeneity of ddl kinetics among healthy individuals. Our work expands on a previous reported method known as RGC. METHODS: Repeated measurements of ddl concentration in the serum were obtained from 48 healthy adult volunteers enrolled in two bioequivalence study. We use the NONMEM program (Non-linear Mixed Effect Model) to estimate the between- and within-subject variability and the corresponding pharmacokinetic parameters. We assess the genetic contribution to the variability of each pharmacokinetic parameter through the RGC method. RESULTS: Pharmacokinetic parameters, expressed as functions of covariates gender and creatinine clearance (CLCR), were: Oral clearance (CL = 55.1 + 240 * CLCR + 16.6 l/h for male and CL = 55.1 + 240 * CLCR for female), central volume (V2 = 9.82), inter-compartmental clearance (Q = 40.90/h), peripheral volume (V3 = 62.7 + 22.90 for male and V3 = 62.70 for female), absorption rate constant (Ka = 1.51 h(-1)) and duration of the dose administration (D = 0.44 h). The RGC of CL, Q, V3, Ka and D were 0.58, 0.97, 0.60, 0.53 and 0.88, respectively. CONCLUSION: We estimated parameter-specific RGC indices and rank them according to the potential genetic contribution as an explanation for the observed variance. Our study design improved precision by decreasing background noise and, thus, improved the chances that indices such as the RGC are in fact describing genetic variability. PMID- 16249660 TI - HIV-1 subtype C dissemination in southern Brazil. AB - OBJECTIVES: To describe the molecular and epidemiological profile of HIV-1 in patients followed at the University Hospital of Rio Grande, Brazil. DESIGN AND METHODS: A cross-sectional study was conducted from September to December 2002. Plasma viral RNA of 85 patients was extracted and protease and reverse transcriptase genes were polymerase chain reaction-amplified and sequenced. Sequences were subtyped and examined to antiretroviral resistance mutations. Laboratory data and past history of antiretroviral treatment were also collected. RESULTS: Most viruses were either subtype B (42%) or subtype C (45%). No risk behaviour, sexual orientation or laboratory parameter was associated with any specific subtype, but subtype C tended to be more frequently found in women (P = 0.06). The prevalence of subtype C has increased over the HIV/AIDS epidemic, accounting for almost 60% of cases diagnosed in 2002. Intra-subtype genetic distances were smaller in subtype C than in subtype B, suggesting a more recent introduction of the former in the epidemic. Of patients under treatment, 60% had at least one antiretroviral drug resistance mutation, but no mutation was specifically associated with any HIV-1 subtype. Only one resistance mutation each was found in drug-naive patients with subtypes B and C. CONCLUSION: Despite the fact that subtype C appeared in southern Brazil more recently than subtype B, it is now the predominant strain in Rio Grande. The epidemic spread of subtype C could be taking place in Brazil, and possibly in south America, a phenomenon similar to that seen in other countries where this subtype is now totally dominant. PMID- 16249662 TI - Does memory priming during anesthesia matter? PMID- 16249663 TI - Continuous peripheral nerve blocks: fewer excuses. PMID- 16249664 TI - The academic highway between the United States and Japan. PMID- 16249665 TI - Investigation of implicit memory during isoflurane anesthesia for elective surgery using the process dissociation procedure. AB - BACKGROUND: This prospective study evaluated memory function during general anesthesia for elective surgery and its relation to depth of hypnotic state. The authors also compared memory function in anesthetized and nonanesthetized subjects. METHODS: Words were played for 70 min via headphones to 48 patients (aged 18-70 yr) after induction of general anesthesia for elective surgery. Patients were unpremedicated, and the anesthetic regimen was free. The Bispectral Index (BIS) was recorded throughout the study. Within 36 h after the word presentation, memory was assessed using an auditory word stem completion test with inclusion and exclusion instructions. Memory performance and the contribution of explicit and implicit memory were calculated using the process dissociation procedure. The authors applied the same memory task to a control group of nonanesthetized subjects. RESULTS: Forty-seven patients received isoflurane, and one patient received propofol for anesthesia. The mean (+/- SD) BIS was 49 +/- 9. There was evidence of memory for words presented during light (BIS 61-80) and adequate anesthesia (BIS 41-60) but not during deep anesthesia (BIS 21-40). The process dissociation procedure showed a significant implicit memory contribution but not reliable explicit memory contribution (mean explicit memory scores 0.05 +/- 0.14, 0.04 +/- 0.09, and 0.05 +/- 0.14; mean automatic influence scores 0.14 +/- 0.12, 0.17 +/- 0.17, and 0.18 +/- 0.21 at BIS 21-40, 41 60, and 61-80, respectively). Compared with anesthetized patients, the memory performance of nonanesthetized subjects was better, with a higher contribution by explicit memory and a comparable contribution by implicit memory. CONCLUSION: During general anesthesia for elective surgery, implicit memory persists even in adequate hypnotic states, to a comparable degree as in nonanesthetized subjects. PMID- 16249666 TI - Detection of consciousness by electroencephalogram and auditory evoked potentials. AB - BACKGROUND: A set of electroencephalographic and auditory evoked potential (AEP) parameters should be identified that allows separation of consciousness from unconsciousness (reflected by responsiveness/unresponsiveness to command). METHODS: Forty unpremedicated patients received anesthesia with remifentanil and either sevoflurane or propofol. With remifentanil infusion (0.2 microg . kg . min), patients were asked every 30 s to squeeze the investigator's hand. Sevoflurane or propofol was given until loss of consciousness. After intubation, propofol or sevoflurane was stopped until patients followed the command (return of consciousness). Thereafter, propofol or sevoflurane was started again (loss of consciousness), and surgery was performed. Return of consciousness was observed after surgery. The electroencephalogram and AEP from immediately before and after the transitions were selected. Logistic regression was calculated to identify models for the separation between consciousness and unconsciousness. For the top 10 models, 1,000-fold cross-validation was performed. Backward variable selection was applied to identify a minimal model. Prediction probability was calculated. The digitized electroencephalogram was replayed, and the Bispectral Index was measured and accordingly analyzed. RESULTS: The best full model (prediction probability 0.89) contained 15 AEP and 4 electroencephalographic parameters. The best minimal model (prediction probability 0.87) contained 2 AEP and 2 electroencephalographic parameters (median frequency of the amplitude spectrum from 8-30 Hz and approximate entropy). The prediction probability of the Bispectral Index was 0.737. CONCLUSIONS: A combination of electroencephalographic and AEP parameters can be used to differentiate between consciousness and unconsciousness even in a very challenging data set. The minimal model contains a combination of AEP and electroencephalographic parameters and has a higher prediction probability than Bispectral Index for the separation between consciousness and unconsciousness. PMID- 16249667 TI - High-frequency components of auditory evoked potentials are detected in responsive but not in unconscious patients. AB - BACKGROUND: The dose-dependent suppression of midlatency auditory evoked potentials by general anesthetics has been proposed to measure depth of anesthesia. In this study, perioperatively recorded midlatency auditory evoked potentials were analyzed in a time-frequency space to identify significant changes induced by general anesthesia. METHODS: Perioperatively recorded auditory evoked potentials of 19 patients, recorded at varying levels of anesthesia, were submitted to a multiscale analysis using the wavelet analysis. Energy contents of the signal were calculated in frequency bands 0-57.1 Hz, 57.1-114.3 Hz, 114.3 228.6 Hz, and 228.6-457.1 Hz. A Friedman test and a Dunn multiple comparisons test were performed to identify significant differences. RESULTS: Statistical evaluation showed a highly significant decrease of the wavelet energies for the frequency bands 57.1-114.3 Hz (P < 0.0001), 114.3-228.6 Hz (P < 0.0001), and 228.6-457.1 Hz (P < 0.0001) for the measuring points representing deep general anesthesia. This decrease is accompanied by a decrease in the wavelet energy of the frequency band 0-57.1 Hz of no statistical significance (P = 0.021) (level of significance set to P = 0.01). The changes are most prominent in the poststimulus interval between 10 and 30 ms. CONCLUSIONS: This study describes the presence of high-frequency components of the auditory evoked potential. The amount of these components is higher during responsiveness when compared to unconsciousness. Temporal localization of the high-frequency components within the auditory evoked potential shows that they represent a response to the auditory stimulus. Further studies are required to identify the source of these high-frequency components. PMID- 16249668 TI - What is the driving performance of ambulatory surgical patients after general anesthesia? AB - BACKGROUND: Ambulatory surgical patients are advised to refrain from driving for 24 h postoperatively. However, currently there is no strong evidence to show that driving skills and alertness have resumed in patients by 24 h after general anesthesia. The purpose of this study was to determine whether impaired driver alertness had been restored to normal by 2 and 24 h after general anesthesia in patients who underwent ambulatory surgery. METHODS: Twenty patients who underwent left knee arthroscopic surgery were studied. Their driving simulation performance, electroencephalographically verified parameters of sleepiness, subjective assessment of sleepiness, fatigue, alertness, and pain were measured preoperatively and 2 and 24 h postoperatively. The same measurements were performed in a matched control group of 20 healthy individuals. RESULTS: Preoperatively, patients had significantly higher attention lapses and lower alertness levels versus normal controls. Significantly impaired driving skills and alertness, including longer reaction time, higher occurrence of attention lapses, and microsleep intrusions, were found 2 h postoperatively versus preoperatively. No significantly differences were found in any driving performance parameters or electroencephalographically verified parameters 24 h postoperatively versus preoperatively. CONCLUSIONS: Patients showed lower alertness levels and impaired driving skills preoperatively and 2 h postoperatively. Based on driving simulation performance and subjective assessments, patients are safe to drive 24 h after general anesthesia. PMID- 16249669 TI - Oxygen and carbon dioxide in the cerebral circulation during progression to brain death. AB - BACKGROUND: The authors propose that for a moderate reduction of perfusion during progressive irreversible ischemia, oxygen extraction increases to maintain aerobic metabolism, and arteriojugular oxygen difference (AJDo2) increases. Because of reduced carbon dioxide washout, venoarterial difference in carbon dioxide tension (DPco2) increases, with no change in the DPco2/AJDo2 ratio. With further reduction of cerebral perfusion, the aerobic metabolism will begin to decrease, AJDo2 will decrease while DPco2 will continue to increase, and the ratio will increase. When brain infarction develops, the metabolism will be abated, no oxygen will be consumed, and no carbon dioxide will be produced. METHODS: The authors studied 12 patients with acute cerebral damage that evolved to brain death and collected intermittent arterial and jugular blood samples. RESULTS: Four patterns were observed: (1) AJDo2 of 4.1 +/- 0.7 vol%, DPco2 of 6.5 +/- 1.9 mmHg, and a ratio of 1.55 +/- 0.3 with cerebral perfusion pressure of 62.5 +/- 13.4 mmHg; (2) a coupled increase of AJDo2 (5.8 +/- 0.7 vol%) and DPco2 (10.1 +/- 1.0 mmHg) with no change in ratio (1.92 +/- 0.14) and cerebral perfusion pressure (57.9 +/- 5.8 mmHg); (3) AJDo2 of 4.7 +/- 0.4 vol% with an increase in DPco2 (11.8 +/- 1 mmHg) and correspondingly higher ratio (2.7 +/- 0.2); in this phase, cerebral perfusion pressure was 39.7 +/- 10.5 mmHg; (4) immediately before diagnosis of brain death (cerebral perfusion pressure, 17 +/- 10.4 mmHg), there was a decrease of AJDo2 (1.1 +/- 0.1 vol%) and of DPco2 (5.3 +/ 0.6 mmHg) with a further ratio increase (5.1 +/- 0.8). CONCLUSIONS: Until compensatory mechanisms are effective, AJDo2 and DPco2 remain coupled. However, when the brain's ability to compensate for reduced oxygen delivery is exceeded, the ratio of DPco2 to AJDo2 starts to increase. PMID- 16249670 TI - Does the investing layer of the deep cervical fascia exist? AB - BACKGROUND: The placement of the superficial cervical plexus block has been the subject of controversy. Although the investing cervical fascia has been considered as an impenetrable barrier, clinically, the placement of the block deep or superficial to the fascia provides the same effective anesthesia. The underlying mechanism is unclear. The aim of this study was to investigate the three-dimensional organization of connective tissues in the anterior region of the neck. METHODS: Using a combination of dissection, E12 sheet plastination, and confocal microscopy, fascial structures in the anterior cervical triangle were examined in 10 adult human cadavers. RESULTS: In the upper cervical region, the fascia of strap muscles in the middle and the fasciae of the submandibular glands on both sides formed a dumbbell-like fascia sheet that had free lateral margins and did not continue with the sternocleidomastoid fascia. In the lower cervical region, no single connective tissue sheet extended directly between the sternocleidomastoid muscles. The fascial structure deep to platysma in the anterior cervical triangle comprised the strap fascia. CONCLUSIONS: This study provides anatomical evidence to indicate that the so-called investing cervical fascia does not exist in the anterior triangle of the neck. Taking the previous reports together, the authors' findings strongly suggest that deep potential spaces in the neck are directly continuous with the subcutaneous tissue. PMID- 16249671 TI - Effects of dexmedetomidine on hippocampal focal adhesion kinase tyrosine phosphorylation in physiologic and ischemic conditions. AB - BACKGROUND: Dexmedetomidine is a potent and selective alpha2-adrenoceptor agonist that exhibits a broad pattern of actions, including sedation, analgesia, and neuroprotection. Some of these actions (e.g., neuroprotection) may require targets involved in long-term cellular changes. The authors hypothesized that dexmedetomidine increases the expression of active (autophosphorylated) focal adhesion kinase (FAK), a nonreceptor tyrosine kinase playing a pivotal role in cellular plasticity and survival. Therefore, we examined the cellular mechanisms involved in this effect and its sensitivity to oxygen-glucose deprivation (OGD) in rat hippocampal slices. METHODS: The effects of dexmedetomidine on phospho tyrosine FAK phosphorylation were studied first with or without various pharmacologic agents in normoxic conditions, and second in a model of pharmacologic preconditioning of slices subjected to 30 min of OGD followed by 1 h of reperfusion. FAK phosphorylation and caspase-3 activation were examined by immunoblotting. Neuronal death was assessed by propidium iodide fluorescence. RESULTS: Dexmedetomidine produced a dose-related increase in FAK phosphorylation (187 +/- 4%, mean +/- SD, from basal level, EC50 = 0.2 microm; 95% confidence interval, 0.09-0.5 microm). This effect was stereoselective and was completely blocked by yohimbine and the combination of the cyclic monophosphate permeant analog 8 bromo cyclic monophosphate and the phosphodiesterase inhibitor 3 isobutyl-1-methylxanthine. It was mimicked by the protein kinase A inhibitor H 89. In contrast, prazosin and the protein kinase C inhibitors chelerythrine and bisindolylmaleimide I were ineffective. OGD induced a significant increase in immunoreactivity of the cleaved caspase-3 17-kd fragment (417 +/- 22; P < 0.001), a decrease in FAK phosphorylation (78 +/- 12% of control; P < 0.05), and production of significant neuronal death. In OGD conditions, a preconditioning application of dexmedetomidine (0.2 microm, 20-min application, 3 h before anoxia) significantly reduced neuronal death and cleaved caspase-3 expression and significantly attenuated the decrease in phosphorylated FAK content. The dexmedetomidine-induced reduction in caspase-3 expression was significantly decreased by the Src tyrosine kinase inhibitor PP2. CONCLUSION: Dexmedetomidine exhibits a preconditioning effect against ischemic injury in hippocampal slices subjected to OGD. Increase in phosphorylation of FAK via stimulation of alpha2 adrenoceptors and decrease in cleaved caspase-3 expression correlate with dexmedetomidine-induced cell survival. PMID- 16249672 TI - Effect of nutritional status on oxidative stress in an ex vivo perfused rat liver. AB - BACKGROUND: Normothermic ischemia-reperfusion is a determinant in liver injury occurring during surgical procedures, ischemic state, and multiple organ failure. The preexisting nutritional status of the liver might contribute to the extent of tissue injury and primary nonfunction. The aim of this study was to determine the role of starvation on hepatic ischemia-reperfusion injury in normal rat livers. METHODS: Rats were randomly divided into two groups: one had free access to food, the other was fasted for 16 h. The portal vein was cannulated, and the liver was removed and perfused in a closed ex vivo system. Two modes of perfusion were applied in each series of rats, fed and fasting. In the ischemia-reperfusion mode, the experiment consisted of perfusion for 15 min, warm ischemia for 60 min, and reperfusion during 60 min. In the nonischemia mode, perfusion was maintained during the 135-min study period. Five rats were included in each experimental condition, yielding a total of 20 rats. Liver enzymes, potassium, glucose, lactate, free radicals, i.e., dienes and trienes, and cytochrome c were analyzed in perfusate samples. The proportion of glycogen in hepatocytes was determined in tissue biopsies. RESULTS: Transaminases, lactate dehydrogenase, potassium, and free radical concentrations were systematically higher in fasting rats in both conditions, with and without ischemia. Cytochrome c was higher after reperfusion in the fasting rats. Glucose and lactate concentrations were greater in the fed group. The glycogen content decreased in both groups during the experiment but was markedly lower in the fasting rats. CONCLUSIONS: In fed rats, liver injury was moderate, whereas hepatocytes integrity was notably impaired both after continuous perfusion and warm ischemia in fasting animals. Reduced glycogen store in hepatocytes may explain reduced tolerance. PMID- 16249673 TI - Isoflurane postconditioning prevents opening of the mitochondrial permeability transition pore through inhibition of glycogen synthase kinase 3beta. AB - BACKGROUND: Postischemic administration of volatile anesthetics activates reperfusion injury salvage kinases and decreases myocardial damage. However, the mechanisms underlying anesthetic postconditioning are unclear. METHODS: Isolated perfused rat hearts were exposed to 40 min of ischemia followed by 1 h of reperfusion. Anesthetic postconditioning was induced by 15 min of 2.1 vol% isoflurane (1.5 minimum alveolar concentration) administered at the onset of reperfusion. In some experiments, atractyloside (10 microm), a mitochondrial permeability transition pore (mPTP) opener, and LY294002 (15 microm), a phosphatidylinositol 3-kinase inhibitor, were coadministered with isoflurane. Western blot analysis was used to determine phosphorylation of protein kinase B/Akt and its downstream target glycogen synthase kinase 3beta after 15 min of reperfusion. Myocardial tissue content of nicotinamide adenine dinucleotide served as a marker for mPTP opening. Accumulation of MitoTracker Red 580 (Molecular Probes, Invitrogen, Basel, Switzerland) was used to visualize mitochondrial function. RESULTS: Anesthetic postconditioning significantly improved functional recovery and decreased infarct size (36 +/- 1% in unprotected hearts vs. 3 +/- 2% in anesthetic postconditioning; P < 0.05). Isoflurane mediated protection was abolished by atractyloside and LY294002. LY294002 inhibited isoflurane-induced phosphorylation of protein kinase B/Akt and glycogen synthase kinase 3beta and opened mPTP as determined by nicotinamide adenine dinucleotide measurements. Atractyloside, a direct opener of the mPTP, did not inhibit phosphorylation of protein kinase B/Akt and glycogen synthase kinase 3beta by isoflurane but reversed isoflurane-mediated cytoprotection. Microscopy showed accumulation of the mitochondrial tracker in isoflurane-protected functional mitochondria but no staining in mitochondria of unprotected hearts. CONCLUSIONS: Anesthetic postconditioning by isoflurane effectively protects against reperfusion damage by preventing opening of the mPTP through inhibition of glycogen synthase kinase 3beta. PMID- 16249674 TI - Neutrophil stimulation with granulocyte colony-stimulating factor worsens ventilator-induced lung injury and mortality in rats. AB - BACKGROUND: Based on the association between the neutrophil and ventilator induced lung injury, the authors hypothesized that neutrophil inhibition with fucoidin would be beneficial and stimulation with granulocyte colony-stimulating factor (G-CSF) would be harmful in a rat model of lethal ventilator-induced lung injury. METHODS: Animals (n = 111) were randomly assigned to be pretreated with fucoidin, G-CSF, or placebo (control) before 4 h of low-tidal-volume (10 ml/kg) or high-tidal-volume (40 ml/kg) mechanical ventilation. RESULTS: All low-volume animals survived. With high volumes, compared with controls, fucoidin did not improve survival (3 of 20 control animals and 5 of 20 fucoidin animals died; P = 0.51) but G-CSF significantly worsened it (18 of 22 animals died; P < 0.001). Circulating neutrophils were increased early with G-CSF and late with fucoidin with low and high tidal volumes (P < 0.05 for each treatment and tidal volume). Fucoidin decreased lung neutrophils, but these were only significant with high tidal volumes, whereas G-CSF increased lung neutrophils but only significantly with low tidal volumes (P < or = 0.01 for each). Fucoidin did not alter any cardiopulmonary measure significantly. Compared with control, G-CSF increased airway pressures with high tidal volumes and worsened lung edema and arterial oxygen with both tidal volumes (P < 0.05 for each). CONCLUSIONS: In this model, neutrophil stimulation by G-CSF increased lung dysfunction and with high tidal volumes worsened survival rates. Extrapolated clinically, neutrophil stimulation either by agents such as G-CSF or conditions such as sepsis may aggravate ventilator-induced lung injury. PMID- 16249675 TI - Isoflurane inhibits cardiac myocyte apoptosis during oxidative and inflammatory stress by activating Akt and enhancing Bcl-2 expression. AB - BACKGROUND: Volatile anesthetics attenuate apoptosis. The underlying mechanisms remain undefined. The authors tested whether isoflurane reduces apoptosis in cardiomyocytes subjected to oxidative or inflammatory stress by enhancing Akt and B-cell lymphoma-2 (Bcl-2). METHODS: Adult and neonatal rat ventricular myocytes and atrial HL-1 myocytes were exposed to hypoxia, hydrogen peroxide, or neutrophils with or without isoflurane pretreatment. The authors assessed cell damage and investigated apoptosis using mitochondrial cytochrome c release, caspase activity, and TUNEL assay. They determined expression of phospho-Akt and Bcl-2 and tested their involvement by blocking phospho-Akt with wortmannin and Bcl-2 with HA14-1. RESULTS: Isoflurane significantly reduced the cell damage and apoptosis induced by hypoxia, H2O2, and neutrophils. Isoflurane reduced hypoxia induced mitochondrial cytochrome c release in HL-1 cells by 45 +/- 12% and caspase activity by 28 +/- 4%; in neonatal cells, it reduced caspase activity by 43 +/- 5% and TUNEL-positive cells by 50 +/- 2%. Isoflurane attenuated H2O2 induced caspase activity in HL-1 cells by 48 +/- 16% and TUNEL-positive cells by 78 +/- 3%; in neonatal cells, it reduced caspase activity by 30 +/- 3% and TUNEL positive cells by 32 +/- 7%. In adult cardiomyocytes exposed to neutrophils, isoflurane decreased both mitochondrial cytochrome c and caspase activity by 47 +/- 3% and TUNEL-positive cells by 25 +/- 4%. Isoflurane enhanced phospho-Akt and Bcl-2 expression. Wortmannin and HA14-1 prevented the action of isoflurane (53 +/ 8% and 54 +/- 7% apoptotic cells vs. 18 +/- 1% without blockers). CONCLUSIONS: Isoflurane protects cardiomyocytes against apoptosis induced by hypoxia, H2O2, or activated neutrophils through Akt activation and increased Bcl-2 expression. This suggests that a reduction in apoptosis contributes to the cardioprotective effects of isoflurane. PMID- 16249676 TI - Effect of halothane on galphai-3 and its coupling to the M2 muscarinic receptor. AB - BACKGROUND: Halothane is an effective bronchodilator and inhibits airway smooth muscle contraction in part by inhibiting intracellular signaling pathways activated by the M2 muscarinic receptor and its cognate inhibitory heterotrimeric guanosine-5'-triphosphate (GTP)-binding protein (G protein), Gi. This study hypothesized that halothane inhibits nucleotide exchange at the alpha isoform-3 subunit of Gi (Galphai-3), but only when regulated by the M2 muscarinic receptor. METHODS: GTP hydrolysis by Galphai-3 and the Galphai-3beta1gamma2HF heterotrimer expressed in Spodoptera frugiperda cells was measured using a phosphohydrolase assay with [gammaPi]-labeled GTP. Anesthetic binding to Galphai-3 was measured by saturation transfer difference nuclear magnetic resonance spectroscopy. Galphai-3 nucleotide exchange was measured in crude membranes prepared from COS-7 cells transiently coexpressing the M2 muscarinic receptor and Galphai-3. A radioactive analog of GTP, [S]GTPgammaS, was used as a reporter for Galphai-3 nucleotide exchange. RESULTS: Although spectroscopy demonstrated halothane binding to Galphai-3, this binding had no effect on [gammaPi]-labeled GTP hydrolysis by the Galphai-3beta1gamma2HF heterotrimer expressed in Spodoptera frugiperda cells, nor basal Galphai-3 nucleotide exchange measured in crude membranes when the muscarinic receptor agonist acetylcholine was omitted from the assay. Conversely, halothane caused a concentration-dependent inhibition of Galphai-3 nucleotide exchange with acetylcholine included in the assay. CONCLUSION: These data indicate that despite halothane binding to Galphai-3, halothane has no direct inhibitory effect on the intrinsic activity of the Galphai-3beta1gamma2HF heterotrimer but inhibits M2 muscarinic receptor regulation of the heterotrimer. This novel effect is consistent with the ability of halothane to inhibit airway smooth muscle contraction and bronchoconstriction induced by acetylcholine. PMID- 16249677 TI - Experimental conditions are important determinants of cardiac inotropic effects of propofol. AB - BACKGROUND: The rationale for this study is that the depressant effect of propofol on cardiac function in vitro is highly variable but may be explained by differences in the temperature and stimulation frequency used for the study. Both temperature and stimulation frequency are known to modulate cellular mechanisms that regulate intracellular free Ca2+ concentration ([Ca2+]i) and myofilament Ca2+ sensitivity in cardiac muscle. The authors hypothesized that temperature and stimulation frequency play a major role in determining propofol-induced alterations in [Ca2+]i and contraction in individual, electrically stimulated cardiomyocytes and the function of isolated perfused hearts. METHODS: Freshly isolated myocytes were obtained from adult rat hearts, loaded with fura-2, and placed on the stage of an inverted fluorescence microscope in a temperature regulated bath. [Ca2+]i and myocyte shortening were simultaneously measured in individual cells at 28 degrees or 37 degrees C at various stimulation frequencies (0.3, 0.5, 1, 2, and 3 Hz) with and without propofol. Langendorff perfused hearts paced at 180 or 330 beats/min were used to assess the effects of propofol on overall cardiac function. RESULTS: At 28 degrees C (hypothermic) and, to a lesser extent, at 37 degrees C (normothermic), increasing stimulation frequency increased peak shortening and [Ca2+]i. Times to peak shortening and rate of relengthening were more prolonged at 28 degrees C compared with 37 degrees C at low stimulation frequencies (0.3 Hz), whereas the same conditions for [Ca2+]i were not altered by temperature. At 0.3 Hz and 28 degrees C, propofol caused a dose-dependent decrease in peak shortening and peak [Ca2+]i. These changes were greater at 28 degrees C compared with 37 degrees C and involved activation of protein kinase C. At a frequency of 2 Hz, there was a rightward shift in the dose response relation for propofol on [Ca2+]i and shortening at both 37 degrees and 28 degrees C compared with that observed at 0.3 Hz. In Langendorff perfused hearts paced at 330 beats/min, clinically relevant concentrations of propofol decreased left ventricular developed pressure, with the effect being less at 28 degrees C compared with 37 degrees C. In contrast, only a supraclinical concentration of propofol decreased left ventricular developed pressure at 28 degrees C at either stimulation frequency. CONCLUSION: These results demonstrate that temperature and stimulation frequency alter the inhibitory effect of propofol on cardiomyocyte [Ca2+]i and contraction. In isolated cardiomyocytes, the inhibitory effects of propofol are more pronounced during hypothermia and at higher stimulation frequencies and involve activation of protein kinase C. In Langendorff perfused hearts at constant heart rate, the inhibitory effects of propofol at clinically relevant concentrations are more pronounced during normothermic conditions. PMID- 16249679 TI - Dural puncture with a 27-gauge Whitacre needle as part of a combined spinal epidural technique does not improve labor epidural catheter function. AB - BACKGROUND: This prospective, double-blind, randomized study was designed to examine whether the combined spinal-epidural technique without subarachnoid drug administration improved epidural catheter function when compared with the traditional epidural technique. METHODS: After institutional review board approval and informed consent, 251 healthy laboring parturients were randomly assigned to either group DP (combined spinal-epidural technique with 27-gauge Whitacre needle dural puncture but without subarachnoid drug administration) or group NoDP (traditional epidural technique). Patient-controlled epidural analgesia was initiated with 0.11% bupivacaine and 2 microg/ml fentanyl. Top-up doses in 5-ml increments of 0.25% bupivacaine were administered if needed. Previous power analysis revealed that a sample size of 108 patients/group was needed to show a clinically useful reduction of the catheter manipulation rate from 32% to 15%. RESULTS: In groups DP and NoDP, 107 and 123 evaluable patients, respectively, completed the study. Demographics and outcome variables measured, including epidural catheter manipulation and replacement rate, sacral sparing, unilateral block, number of top-up doses, average hourly epidural drug usage, highest sensory blockade level, and labor analgesia quality, were not different between groups. A subgroup of 18 patients without cerebral spinal fluid return during dural puncture had a higher catheter replacement rate than those of groups DP and NoDP, but it did not reach statistical significance. CONCLUSIONS: Dural puncture with a 27-gauge Whitacre needle without subarachnoid drug administration during combined spinal-epidural labor analgesia did not improve epidural labor analgesia quality or reduce catheter manipulation or replacement rate when compared with a traditional epidural technique. PMID- 16249678 TI - Continuous peripheral nerve blocks in hospital wards after orthopedic surgery: a multicenter prospective analysis of the quality of postoperative analgesia and complications in 1,416 patients. AB - BACKGROUND: Continuous peripheral nerve block (CPNB) is the technique of choice for postoperative analgesia after painful orthopedic surgery. However, the incidence of neurologic and infectious adverse events in the postoperative period are not well established. This issue was the aim of the study. METHODS: Patients scheduled to undergo orthopedic surgery performed with a CPNB were prospectively included during 1 yr in a multicenter study. Efficacy of postoperative analgesia, bacteriologic cultures of the catheter, and acute neurologic and infectious adverse events were evaluated after surgery in 1,416 patients at arrival in the postanesthesia care unit, at hour 1, and every 24 h up to day 5. Risk factors for adverse events were determined using logistic regression. RESULTS: The median duration of CPNB was 56 h. Both general anesthesia and CPNB were performed in 73.6% of the patients. Postoperative analgesia was effective in 96.3%, but an increase in pain scores was noted at hour 24 (P = 0.01). Hypoesthesia or numbness occurred in 3% and 2.2%, respectively, and paresthesia occurred in 1.5%. Three neural lesions (0.21%) were noted after continuous femoral nerve block. Two of these patients were anesthetized during block procedure. Nerve damage completely resolved 36 h to 10 weeks later. Cultures from 28.7% of the catheters were positive. Three percent of patients had local inflammatory signs. The bacterial species most frequently found were coagulase-negative staphylococcus (61%) and gram-negative bacillus (21.6%). A Staphylococcus aureus psoas abscess (0.07%) was reported in one diabetic woman. Independent risk factors for paresthesia/dysesthesia were postoperative monitoring in intensive care, age less than 40 yr, and use of bupivacaine. Risk factors for local inflammation/infection were postoperative monitoring in intensive care, catheter duration greater than 48 h, male sex, and absence of antibiotic prophylaxis. CONCLUSION: CPNB is an effective technique for postoperative analgesia. Minor incidents and bacterial colonization of catheters are frequent, with no adverse clinical consequences in the large majority of cases. Major neurologic and infectious adverse events are rare. PMID- 16249680 TI - Physiologic and antinociceptive effects of intrathecal resiniferatoxin in a canine bone cancer model. AB - BACKGROUND: Resiniferatoxin is a potent capsaicin analog. Intrathecal administration leads to selective, prolonged opening of the transient receptor potential V1 ion channel, which is localized mainly to C-fiber primary afferent nociceptive sensory neurons. Following work in laboratory animals, the authors explored the use of intrathecal resiniferatoxin to control spontaneous bone cancer pain in companion (pet) dogs. METHODS: Normal canine population: Behavioral testing was performed to establish baseline paw withdrawal latency; subsequently, general anesthesia was induced and resiniferatoxin was administered intrathecally while hemodynamic parameters were recorded. Behavior testing was repeated for 12 days after administration of resiniferatoxin. Clinical canine population: Twenty companion dogs with bone cancer pain were recruited. The animal's baseline level of discomfort and analgesic use were recorded. Resiniferatoxin was administered intrathecally and hemodynamic parameters were monitored while the dogs were under general anesthesia. Dogs were reevaluated up to 14 weeks after resiniferatoxin administration. RESULTS: Normal canine population: In the first minutes after resiniferatoxin injection, there were significant (P < 0.05) increases in mean arterial blood pressure and heart rate from baseline. Two days after injection, limb withdrawal latencies increased to the point of cutoff in the dogs that received at least 1.2 microg/kg resiniferatoxin. Clinical canine population: From baseline, there were significant (P < 0.05) increases in mean arterial blood pressure and heart rate after resiniferatoxin injection. Comfort scores were significantly improved at 2, 6, 10, and 14 weeks after resiniferatoxin administration (P < 0.0001). There was decreased or discontinued use of supplemental analgesics in 67% of the dogs 2 weeks after resiniferatoxin administration. CONCLUSIONS: Intrathecal resiniferatoxin elicits transient hemodynamic effects. In controls, a profound and sustained blockade of thermal stimuli is produced in a dose-dependent fashion. Similar administration in dogs with bone cancer produces a prolonged antinociceptive response. PMID- 16249681 TI - Adenosine reduces glutamate release in rat spinal synaptosomes. AB - BACKGROUND: A1 adenosine receptor activation reduces hypersensitivity in animal models of chronic pain, but intrathecal adenosine does not produce analgesia to acute noxious stimuli. Here, the authors test whether increased inhibition by adenosine of glutamate release from afferents after injury accounts for this difference. METHODS: Synaptosomes were prepared from the dorsal half of the lumbar spinal cord of normal rats or those with spinal nerve ligation. Glutamate release evoked by the TRPV-1 receptor agonist, capsaicin, was measured. Adenosine with or without adenosine A1 and A2 receptor antagonists was applied to determine the efficacy and mechanism of adenosine to reduce capsaicin-evoked glutamate release. RESULTS: Capsaicin produced a concentration-dependent glutamate release similarly in normal and nerve-injured rats. Capsaicin-evoked glutamate release was inhibited by adenosine or R-PIA (R-N6-(2- phenylisopropyl)-adenosine) in a concentration-dependent manner, with a threshold of 10 nm in both normal and nerve-ligated synaptosomes. Blockade of capsaicin-evoked glutamate release by adenosine was reversed similarly in synaptosomes from normal and spinal nerve ligated animals by an A1 adenosine receptor antagonist DPCPX (8-cyclopentyl-1,3 dipropylxanthine) but not by an A2 adenosine receptor antagonist DMPX (3'7 dimethyl-1-proparaglyxanthine). Capsaicin-evoked glutamate release, as well as its inhibition by adenosine, did not differ between synaptosomes prepared from tissue ipsilateral and contralateral to spinal nerve ligation. CONCLUSION: These observations confirm previous neurophysiologic studies that presynaptic adenosine A1 receptor activation inhibits glutamate release from primary afferents. This effect is unaltered after peripheral nerve injury and thereby is unlikely to account for the enhanced analgesic efficacy of intrathecal adenosine in this setting. PMID- 16249682 TI - Receptors, G proteins, and their interactions. AB - Membrane receptors coupling to intracellular G proteins (G protein-coupled receptors) form one of the major classes of membrane signaling proteins. They are of great importance to the practice of anesthesiology because they are involved in many systems of relevance to the specialty (cardiovascular and respiratory control, pain transmission, and others) and many drugs target these systems. In recent years, understanding of these signaling systems has grown. The structure of receptors and G proteins has been elucidated in more detail, their regulation is better understood, and the complexity of interactions between the various parts of the system (receptors, G proteins, effectors, and regulatory molecules) has become clear. These findings may help explain both actions and side effects of drugs. In addition, these newly discovered targets are likely to play important roles in disease states of relevance to anesthesiologists. PMID- 16249683 TI - Efficacy of postoperative patient-controlled and continuous infusion epidural analgesia versus intravenous patient-controlled analgesia with opioids: a meta analysis. AB - The authors performed a meta-analysis and found that epidural analgesia overall provided superior postoperative analgesia compared with intravenous patient controlled analgesia. For all types of surgery and pain assessments, all forms of epidural analgesia (both continuous epidural infusion and patient-controlled epidural analgesia) provided significantly superior postoperative analgesia compared with intravenous patient-controlled analgesia, with the exception of hydrophilic opioid-only epidural regimens. Continuous epidural infusion provided statistically significantly superior analgesia versus patient-controlled epidural analgesia for overall pain, pain at rest, and pain with activity; however, patients receiving continuous epidural infusion had a significantly higher incidence of nausea-vomiting and motor block but lower incidence of pruritus. In summary, almost without exception, epidural analgesia, regardless of analgesic agent, epidural regimen, and type and time of pain assessment, provided superior postoperative analgesia compared to intravenous patient-controlled analgesia. PMID- 16249684 TI - Government account for relief in occupied area: a Japanese physician's journey to a new medical specialty. PMID- 16249685 TI - Successful resuscitation after cardiovascular collapse following accidental intravenous infusion of levobupivacaine during general anesthesia. PMID- 16249686 TI - Massive inhalation of desflurane due to vaporizer dysfunction. PMID- 16249687 TI - Pharmacogenetics of anesthetic and analgesic agents: CYP2D6 genetic variations. PMID- 16249688 TI - The practicality and need for genetic testing for malignant hyperthermia. PMID- 16249691 TI - Interaction between anesthetic molecules and their binding sites must be far more complex. PMID- 16249692 TI - Rigid endoscopy for assessment of extraglottic airway device position. PMID- 16249693 TI - Let there be light! (preferably on the inside): misassembled laryngoscope blade as cause of failed intubation. PMID- 16249694 TI - Effect of transarterial axillary block versus general anesthesia on paresthesiae 1 year after hand surgery. PMID- 16249696 TI - Cross-clade CD8(+) T-cell responses with a preference for the predominant circulating clade. AB - Human immunodeficiency virus (HIV) genetic diversity is a major impediment to the design of a successful vaccine. Even if an HIV vaccine is proven effective, it remains to be seen whether this protection will extend to inter-clade, intra clade, and recombinant strains. We used recombinant vaccinia-based interferon gamma (IFN) Elispot assays to test the inter-clade crossreactivity of clades A, B, C, and D HIV Env in two cohorts of HIV-infected Kenyans. Despite the tremendous diversity in this HIV protein, a substantial proportion of multi-clade responses were observed. Although these multi-clade responses correlated well with each other in regression analyses, clade A responses were seen at a higher frequency and at greater relative magnitudes in a proportion of these patients, when compared to the other three clades. Epitope mapping indicates CD8(+) T cell recognition of conserved regions of Env, accounting for the high degree of cross reactivity but not the clade A preference. A better understanding of cross-clade CD8(+) T cell responses to HIV may help to predict whether a successful vaccine could be used to stop geographically and genetically distinct HIV epidemics. PMID- 16249697 TI - HIV-1 p24 may persist during long-term highly active antiretroviral therapy, increases little during short treatment breaks, and its rebound after treatment stop correlates with CD4(+) T cell loss. AB - The dynamics of HIV-1 RNA during structured treatment interruptions (STIs) are well established, but little is known about viral proteins like p24. We studied 65 participants of an STI trial. Before the trial, continuous highly active antiretroviral therapy (HAART) had suppressed their viral load to <50 copies/mL during 6 months. They then interrupted HAART during weeks 1 through 2, 11 through 12, 21 through 22, 31 through 32, and 41 through 52. The p24 was measured by boosted enzyme-linked immunosorbent assay of plasma pretreated by efficient virus disruption and heat denaturation. At time point 0, p24 was measurable in 22 patients (34%), who had maintained a viral load <50 copies/mL for 25.4 months (median, range: 6.2-38.9 months) under HAART. Viral rebounds during 2-week STIs led to a mean p24 increase of only 0.08 to 0.19 log10 (ie, 20%-60%). Pre-HAART viral load and p24 at time 0 independently predicted p24 rebounds during the 4 2 week STIs. The p24 at time 0 and HIV-1 RNA rebound during weeks 41 through 52 independently determined the concomitant p24 rebound. An increase of p24 but not viral load during the first 8 weeks of the long STI correlated significantly with concomitant CD4(+) T cell loss. Persisting p24 despite successful HAART may reflect virus replication in reservoirs not represented by plasma viral load and has implications for the concept of therapeutic vaccination. PMID- 16249699 TI - Enhanced replication of R5 HIV-1 over X4 HIV-1 in CD4(+)CCR5(+)CXCR4(+) T cells. AB - To enter human cells, HIV-1 usually uses CD4 and 1 of 2 coreceptors: CCR5 and CXCR4. Interestingly, even though CCR5 is expressed on far fewer T cells than is CXCR4, many patients in early- and late-stage HIV disease maintain high levels of CCR5-tropic (R5) viruses. We hypothesized that such high R5 viral loads may be sustained because, relative to CXCR4-tropic (X4) HIV-1 infection, R5 HIV-1 infection of permissive CD4(+)CCR5(+)CXCR4(+) T cells results in the production of significantly more infectious virus particles per target cell. To investigate this possibility, we compared the levels of virus production per target cell after isogenic R5 and X4 HIV-1 infection of 2 in vitro primary human lymphocyte culture systems: T-cell receptor-stimulated blood-derived CD4(+) T cells and tonsil histoculture (which requires no exogenous stimulation for ex vivo infection). We provide evidence that R5 HIV-1 does indeed compensate for a small target cell population by producing, on average, 5 to 10 times more infectious virus per CCR5(+) target cell than X4 HIV-1. This replicative advantage may contribute to the predominance of R5 HIV-1 in vivo. PMID- 16249698 TI - Role of lymphocyte multidrug resistance protein 1 in HIV infection: expression, function, and consequences of inhibition. AB - The multidrug resistance protein 1 (MRP1) is a drug transporter that protects cells from oxidative stress, which increases HIV-1 replication. The aim of this study was to characterize the expression, function, and role of lymphocyte MRP1 in HIV-1 infection and its modulation by antiretroviral drugs such as the protease inhibitors (PIs). Peripheral blood mononuclear cells (PBMCs) from HIV positive individuals do not show significant alterations of MRP1 expression despite highly active antiretroviral therapy and HIV plasma viral load levels; however, they exhibit different intracellular MRP1 expression as compared with healthy subjects. By contrast, MRP efflux function is increased in subjects with primary HIV infection and becomes defective in later stages of the infection. PI- and probenecid (PBCD)-mediated inhibition of MRP lowers the in vitro stress induced response of lymphoid cells by reducing the level of the specific reactive oxygen species superoxide anion and hydrogen peroxide. Finally, the blockade of MRP by PBCD and PIs down-modulates HIV-1 replication by a mechanism independent of inhibition of the HIV-1 protease. Our results are consistent with a model wherein HIV replication is favored by the MRP1-related oxidative stress and inhibition of MRP1 may contribute to the antiviral activity of PIs. PMID- 16249700 TI - Lack of association of SDF-1 3'A variant allele with long-term nonprogressive HIV 1 infection is extended beyond 16 years. AB - We studied the frequency of the SDF-1 3'A allelic variant (801G-->A) in a cohort of white Spaniards made up of (1) HIV-1-infected long-term nonprogressors (LTNPs) older than 16 years of age (n = 57), (2) HIV-1-infected usual progressors (UPs; n = 107), and (3) a group of healthy controls (n = 100). The mutant SDF-1 3'A allele was observed in 28% of LTNPs, 19% of UPs, and 26% of healthy controls (P = not significant). Homozygosity for the 3'A mutation was detected in 7%, 4%, and 3% of LTNPs, UPs, and healthy controls, respectively (P = not significant). Polymorphism at the SDF-1 locus is not associated with LTNP disease of longer than 16 years in Spanish HIV-1-infected patients. This effect is independent of the CCR5Delta32 allele. PMID- 16249701 TI - Lopinavir/ritonavir as single-drug therapy for maintenance of HIV-1 viral suppression: 48-week results of a randomized, controlled, open-label, proof-of concept pilot clinical trial (OK Study). AB - OBJECTIVE: This study evaluated maintenance with lopinavir/ritonavir monotherapy vs. continuing lopinavir/ritonavir and 2 nucleosides in HIV-infected patients with suppressed HIV replication. DESIGN: Randomized, controlled, open-label, multicenter, pilot clinical trial. METHODS: Adult patients were eligible if they had no history of virologic failure while receiving a protease inhibitor, were receiving 2 nucleosides + lopinavir/ritonavir (400/100 mg b.i.d.) for >1 month and had maintained serum HIV RNA <50 copies/mL for >6 months prior to enrollment. RESULTS: Forty-two patients were randomly assigned 1:1 to continue or stop the nucleosides. At baseline there were no significant differences between groups in median CD4 cells/muL (baseline or nadir), pre-HAART (highly active antiretroviral therapy) HIV log10 viremia, or time with HIV RNA <50 copies/mL prior to enrollment. After 48 weeks of follow-up, percentage of patients remaining at <50 HIV RNA copies/mL (intention to treat, M = F) was 81% for the monotherapy group (95% CI: 64% to 98%) vs. 95% for the triple-therapy group (95% CI: 86% to 100%); P = 0.34. Patients in whom monotherapy failed had significantly worse adherence than patients who remained virally suppressed on monotherapy. Monotherapy failures did not show primary resistance mutations in the protease gene and were successfully reinduced with prerandomization nucleosides. Mean change in CD4 cells/microL: +70 (monotherapy) and +8 (triple) (P = 0.27). Mean serum fasting lipids remained stable in both groups. No serious adverse events were observed. CONCLUSION: Most of the patients maintained with lopinavir/ritonavir monotherapy remain with undetectable viral load after 48 weeks. Failures of lopinavir/ritonavir monotherapy were not associated with the development of primary resistance mutations in the protease gene and could be successfully reinduced adding back prior nucleosides. PMID- 16249702 TI - Discordant immunologic and virologic responses to highly active antiretroviral therapy are associated with increased mortality and poor adherence to therapy. AB - OBJECTIVE: To examine the independent association of discordant virologic and immunologic responses to highly active antiretroviral therapy (HAART) with mortality. METHODS: A population-based study of 1527 treatment-naive individuals initiating HAART used Cox proportional hazards modeling to determine the independent association of treatment response at 3 to 9 months with nonaccidental mortality. Logistic regression was used to examine associations with discordant responses. RESULTS: Viral load (VL)/CD4 discordant responses were seen in 235 (15.4%) of subjects, and VL/CD4 responses were seen in 179 (11.7%) of subjects. In adjusted Cox regression models, discordant responses were found to be independently associated with an increased risk of mortality (VL/CD4: relative hazard [RH] = 1.87, 95% confidence interval [CI]: 1.15 to 3.04; VL/CD4: RH = 2.47, 95% CI: 1.54 to 3.95). VL/CD4 discordance was found to be associated with increasing age, baseline HIV RNA load <100,000 copies/mL, baseline CD4 counts <50 cells/muL, the use of lamivudine (3TC)/zidovudine (ZDV), and poor adherence to therapy. VL/CD4 discordance was associated with younger age; injection drug use; baseline HIV RNA load >100,000 copies/mL; the use of 3TC/ZDV, didanosine (ddI)/3TC, or ddI/stavudine; and poor adherence to therapy. CONCLUSION: Discordant responses are independently associated with an increased risk of mortality and are, in turn, associated with poor adherence to therapy. PMID- 16249703 TI - Selective drug taking during combination antiretroviral therapy in an unselected clinic population. AB - OBJECTIVES: Multidrug therapy is necessary to achieve sustained viral suppression. Discordant adherence to individual components of a multidrug regimen may lead to adverse outcomes. METHODS: Antiretroviral-naive patients initiating therapy from 1997 through 2002 were included. Adherence for each antiretroviral was determined using pharmacy refill data. Selective drug taking was defined as > or =5% difference in adherence between 2 components of an antiretroviral regimen lasting at least 60 days. RESULTS: A total of 322 of 415 patients (78%) met inclusion criteria. Selective drug taking occurred in 47 of 322 patients (15%) and on 51 of 438 regimens (12%). Factors associated with selective drug taking were lower baseline CD4 lymphocyte count (adjusted odds ratio [AOR]: 1.3, 95% CI: 1.1 to 1.6 per 100 cell/microL decrease); 3 times daily dosing schedule (AOR: 4.1, 95% CI: 1.1 to 15.5); and the presence of significant adverse drug events (AOR: 2.9, 95% CI: 1.3 to 6.4). Regimens containing a fixed-dose combination dosage form were less likely to have selective drug taking (AOR: 0.5, 95% CI: 0.2 to 0.99). Outcomes independently associated with selective drug taking included earlier progression to a new AIDS-defining illness or death (hazard ratio: 2.3, 95% CI: 1.2 to 4.5). CONCLUSIONS: Selective drug taking was relatively common among patients taking combination antiretroviral therapy. The factor most closely associated with selective drug taking was the presence of an adverse drug event. Clinical outcomes appeared worse in patients with selective drug taking. PMID- 16249704 TI - Four measures of antiretroviral medication adherence and virologic response in AIDS clinical trials group study 359. AB - AIDS Clinical Trials Group (ACTG) 359 was a randomized, partially double-blinded factorial study of 6 antiretroviral regimens, all including saquinavir, among HIV infected persons in whom prior therapy had failed (n = 258). Counts of remaining saquinavir capsules were determined between weeks 0 and 4; at weeks 4, 8, and 16, self-reported adherence was estimated from 2-day report of doses skipped, therapeutic coverage, and percent of doses taken were determined by electronic monitoring devices applied to saquinavir bottles, and the saquinavir 24-hour area under the curve (AUC) was estimated. Relationships were evaluated among these 4 adherence measures and the primary endpoint of week 16 HIV RNA change. Thirty percent of 254 subjects had HIV RNA < or =500 copies/mL at week 16. Only self reported adherence and saquinavir AUC were significantly associated with week 16 HIV RNA change (P = 0.019 and 0.023, respectively), and these measures were higher in subjects with week 16 HIV RNA < or =500 copies/mL (P = 0.03 and 0.008, respectively). The ability to detect a correlation between electronically monitored adherence and virologic response was limited by the small sample size. Self-reported adherence and saquinavir AUC were significant predictors of virologic response, in this evaluation. These findings provide insight into methods of assessing and improving adherence to antiretroviral regimens. PMID- 16249705 TI - Structured treatment interruptions in primary HIV-1 infection: the ANRS 100 PRIMSTOP trial. AB - BACKGROUND: Whether structured treatment interruptions (STIs) can induce anti-HIV immune response and control HIV replication following discontinuation of highly active antiretroviral therapy (HAART) in patients with primary HIV infection is controversial. METHODS: In this multicenter, prospective trial, patients with early symptomatic primary HIV infection were given HAART continuously for 34 weeks. Afterward, patients with plasma viral load (PVL) <50 copies/mL entered the STI phase, which consisted of 3 consecutive periods of 2, 4, and 8 weeks off HAART, each separated by 12 weeks on HAART. HAART was permanently stopped at week 84 and patients were followed up for 24 weeks. The primary endpoint for definition of virologic success was a PVL <50 copies/mL during the 6 months following HAART discontinuation. RESULTS: Of the 29 patients enrolled, 26 completed the trial. Six months after HAART discontinuation, only 1 patient (3.8%, 95% CI: 0.1% to 19.6%) had PVL <50 copies/mL, whereas 6 of 26 (23.1%, 95% CI: 9.0% to 43.7%) had PVL <1000 copies/mL. Female gender was the only parameter significantly associated with a PVL <1000 copies/mL. No other parameter, either at baseline or before HAART discontinuation, predicted virologic success at week 108. A major protease inhibitor resistance mutation (L90M) developed in 3 patients. CONCLUSIONS: This trial failed to confirm that a significant proportion of patients with primary HIV infection can maintain suppression of viremia after a sequence of HAART/STIs followed by HAART discontinuation. PMID- 16249706 TI - Once-daily regimen of saquinavir, ritonavir, didanosine, and lamivudine in HIV infected patients with standard tuberculosis therapy (TBQD Study). AB - OBJECTIVES: To assess the efficacy and safety of a once-daily regimen with didanosine, lamivudine, saquinavir, and low-dose ritonavir in antiretroviral (ARV)-naive patients with tuberculosis treated with rifampin and the influence of rifampin on plasma trough concentration (Ctrough) of saquinavir. METHODS: Single arm, prospective, multicenter, open-label pilot study, including 32 adult ARV naive subjects with HIV infection and tuberculosis under standard treatment that included rifampin (600 mg q.d.) and isoniazid (300 mg q.d.). After 2 months of tuberculosis treatment, patients were started on once-daily ARV therapy, consisting of didanosine, lamivudine, ritonavir (200 mg), and saquinavir soft gel capsules (1600 mg). HIV RNA level, CD4 cell count, clinical and laboratory toxicity, and saquinavir Ctrough during and after antituberculosis therapy were analyzed. RESULTS: After 48 weeks of follow-up, 20 of 32 patients (62.5%; 95% CI: 45.8% to 79.2%) in the intent-to-treat population and 20 of 28 (71.4%; 95% CI: 54.4% to 88.4%) in the on-treatment population had an HIV RNA level <50 copies/mL. Treatment tolerance was acceptable in all patients except for 2 with biologic hepatic toxicity leading to discontinuation. Seven patients had virologic failure. In 10 patients (36%), saquinavir Ctrough was <0.05 microg/mL during tuberculosis therapy and 5 of them had virologic failure. The median saquinavir Ctrough was 44% lower (interquartile range: 19% to 71%) with coadministration of rifampin than without. CONCLUSION: The combination of didanosine, lamivudine, saquinavir, and ritonavir may be a useful treatment regimen for patients with tuberculosis in whom a once-daily protease inhibitor containing regimen is considered indicated. Nevertheless, on the basis of pharmacokinetic profile the dose of 1600/200 mg of saquinavir/ritonavir cannot be recommended. Further studies with higher doses of saquinavir (2000 mg) boosted with ritonavir are warranted. PMID- 16249707 TI - Predictors of virologic failure in HIV-1-infected patients starting highly active antiretroviral therapy in Porto Alegre, Brazil. AB - OBJECTIVE: To assess predictors of virologic response 6 months after initiation of highly active antiretroviral therapy (HAART) in a cohort of HIV-infected patients in Brazil. METHODS: Treatment-naive patients who started HAART between 1996 and 2004 and had information on viral load at 3-9 months were included. Information was collected on demographic characteristics, antiretroviral regimen, adherence, AIDS diagnosis, baseline CD4 cell count, and viral load. Virologic failure (VF) was defined as viral load > or =400 copies/mL at 6 months or death before completion of 6 months of therapy. RESULTS: Among 454 patients who met the inclusion criteria, VF occurred in 127 (28.0%). In univariate analysis, VF was associated with younger age (median 34 vs. 37 years, P = 0.003), AIDS diagnosis (relative risk [RR] 1.18, P = 0.009), higher baseline viral load (5.34 vs. 5.00, P = 0.0002), lower baseline CD4 cell count (86 vs. 182, P = 0.006), nonadherence (RR 1.39, P < 0.0001), regimen containing 1 single protease inhibitor, as compared with ritonavir-boosted regimens (odds ratio [OR] 8.5, P < 0.0001), and year therapy initiated before 1999 (P < 0.0001). To minimize the systematic effect of therapy indication, we analyzed the subset of 158 patients with CD4 count < or =200 cells/microL who started therapy after 1999. After adjusting for age, education, adherence, regimen, and baseline viral load, nonadherence (OR 8.78, P = 0.02), and fewer years of education (OR 6.05, P = 0.05) remained associated with VF. CONCLUSIONS: A significant improvement was found in virologic suppression over time, consistent with the introduction of nonnucleoside reverse transcriptase inhibitors and ritonavir-boosted regimens into clinical practice. With currently available therapies, compliance and education were shown to be predictors of virologic response, particularly in more immunocompromised patients. PMID- 16249708 TI - The role for government health centers in provision of same-day voluntary HIV counseling and testing in Kenya. AB - OBJECTIVE: To explore the role of primary health centers in provision of voluntary counseling and testing (VCT) in Kenya. DESIGN AND SETTING: Prospective service evaluation at 3 (1 urban and 2 rural) government health centers. SUBJECTS: Consecutive adult clients. MAIN OUTCOME MEASURES: Uptake of services, user characteristics, quality of service. RESULTS: Counseling services received 2315 new clients over 26 months. The last quarter averaged 101 clients per clinic. More than 80% of clients lived locally. Overall 93% opted to test, 91% receiving results, 82% on the same day. Most clients tested HIV negative (81%). Youth and men were well represented. Few couples (10%) attended. Seventeen percent of women were pregnant. Self-referral was common and illness was an uncommon reason for testing (<20%). Thirty-one percent of clients were referred from VCT to other health center services. Counseling was perceived as high quality by users and providers. Validation of the test algorithm showed a sensitivity of 98.0% and specificity of 98.7%. CONCLUSION: Government health centers in Kenya can be appropriate providers of VCT. This pilot helped initiate a new strategy of health center-based VCT in Kenya and this has facilitated rapid expansion and more equitable provision for Kenyans. PMID- 16249710 TI - Preventing mother-to-child transmission of HIV in Western Kenya: operational issues. AB - OBJECTIVES: To improve uptake in a program to prevent mother-to-child HIV transmission and describe lessons relevant for prevention of mother-to-child transmission programs in resource-poor settings. METHODS: Implementation of a pilot project that evaluates approaches to increase program uptake at health facility level at New Nyanza Provincial General Hospital, a public hospital in western Kenya, an area with high HIV prevalence. Client flow was revised to integrate counseling, HIV testing, and dispensing of single-dose nevirapine into routine antenatal services. The number of facilities providing PMCT services was expanded to increase district-wide coverage. Main outcome measures were uptake of counseling, HIV testing, nevirapine, and estimated program impact. RESULTS: Uptake of counseling and testing improved from 55 to 68% (P < 0.001), nevirapine uptake from 57% to 70% (P < 0.001), and estimated program impact from 15% to 23% (P = 0.03). Aggregate reports compare well with computer-entered data. CONCLUSION: Addressing institutional factors can improve uptake, but expected program impact remains low for several reasons, including relatively low efficacy of the intervention and missed opportunities in the labor room. PMID- 16249709 TI - Initial response to highly active antiretroviral therapy in HIV-1C-infected adults in a public sector treatment program in Botswana. AB - OBJECTIVE: To describe the response to highly active antiretroviral treatment (HAART) in a public sector pilot antiretroviral (ARV) treatment program in Botswana. METHODS: The response to HAART is described in adult HIV-infected ARV naive patients initiating treatment from April 2001 to January 2002 at Princess Marina Hospital in Gaborone, Botswana. Patients had medical and laboratory evaluations before initiating ARV treatment and were followed longitudinally. For analysis, data were collected from charts and patient management records. RESULTS: One hundred fifty-three ARV-naive patients initiated HAART. Most received didanosine plus stavudine (ddI + d4T) with efavirenz or nevirapine. The mean CD4 cell count increase was 149 cells/mm at 24 weeks and 204 cells/mm at 48 weeks. The percentage of patients with an HIV-1 RNA level < or =400 copies/mL was 87.0% at 24 weeks and 78.8% at 48 weeks. The Kaplan-Meier 1-year survival estimate was 84.7% (79.0%, 90.8%), with a 3.2-fold increased risk (P = 0.004) of mortality among patients with a CD4 cell count <50 cells/mm. The 1-year Kaplan Meier estimate of toxicity-related drug switches was 32.2% (20.3%, 40.4%). The most common toxicity was peripheral neuropathy, occurring more frequently in patients with a preexisting diagnosis of peripheral neuropathy and among those placed on ddI + d4T-containing regimens. CONCLUSIONS: An excellent response to HAART was observed among HIV-1C-infected patients, paralleling those seen elsewhere. Despite excellent responses, high rates of toxicity were observed for ddI + d4T-containing regimens. PMID- 16249711 TI - Cost and enrollment implications of targeting different source population for an HIV treatment program. AB - BACKGROUND: Rapid scale-up of antiretroviral therapy (ART) is a worldwide priority, and ambitious targets for numbers on ART have been set. Antenatal clinics (ANCs) and tuberculosis (TB) clinics have been targeted as entry points into HIV care. METHODS: We developed a conditional probability model to evaluate the effects of ANC and TB clinic populations on ART program enrollment. RESULTS: To start 1 individual on ART, 3 TB patients have to be screened at a crude program cost of 36 US dollars per patient initiated on therapy. By contrast, 48 ANC patients have to be screened at a cost of US 214 US dollars per patient on therapy. In an incremental analysis in which ANC HIV testing was borne by a program to prevent mother-to-child transmission, recruitment efficiency increased (8 screened per patient starting ART) and cost decreased (114 US dollars per patient on therapy). Absolute numbers starting ART, however, remained fixed. If all 60,000 ANC patients seen yearly in the Lusaka District were screened, 1247 would start ART. Approaching the district's 35,000 annual TB patients would generate 11,947 patients on ART. CONCLUSION: In areas with high HIV prevalence, targeting chronically ill populations for HIV treatment may have significant short-term benefits in cost savings and recruitment efficiency. PMID- 16249712 TI - Effect of a brief antiretroviral adherence intervention delivered by HIV care providers. AB - Antiretroviral therapy (ART) is effective in controlling viral load in many people infected with HIV, but high levels of adherence to ART are needed for prolonged viral suppression. This study evaluated a brief adherence intervention delivered to HIV-positive patients by primary care providers during routine medical examinations. Six clinics were randomly allocated to deliver an intervention focusing on ART adherence (2 clinics) or safer sex (4 clinics). Interventions included written information (posters, brochures, and flyers) and brief counseling from providers and were evaluated with cohorts of randomly selected patients (n = 437) measured before and after a 10-month intervention. Among those 95% or greater adherent at baseline, 91% of patients who received the adherence intervention remained 95% or greater adherent at follow-up compared with 75% of the patients who received the safer sex intervention (chi = 12.59, P < 0.01). This difference was significant in a logistic regression analysis (odds ratio = 2.26; 95% confidence interval = 1.27-4.04), adjusting for baseline adherence, demographics, and HIV medical status. The adherence intervention did not significantly increase the prevalence of 95% or greater adherence among patients less than 95% adherent at baseline. Similar but nonsignificant results were observed for viral load. A brief intervention delivered to HIV patients by their primary providers helped to maintain adequate adherence to ART regimens. More intensive intervention is needed to improve adherence among patients who are initially less than 95% adherent. PMID- 16249713 TI - Does exposure to antiretroviral therapy affect growth in the first 18 months of life in uninfected children born to HIV-infected women? AB - Uninfected children born to HIV-infected women are exposed antenatally to antiretroviral therapy, but it is uncertain whether this affects growth in early life. We analyzed weight, height, and occipitofrontal circumference (OFC) in 1912 children from a cohort study: 1304 had no or monotherapy exposure and 608 had combination therapy exposure. The mean z-score for birth weight or OFC did not differ by exposure category in 1513 term children or in 78 born at <34 weeks; the 266 born from 34 to 36 weeks were heavier if exposed to combination therapy. Children with combination therapy exposure born at 34 to 36 weeks reached the 25th centile for weight and OFC earlier than those not exposed born at 34 to 36 weeks (median: birth vs. 3 months; P = 0.003 [weight], P = 0.004 [OFC]), whereas children exposed to combination therapy born at <34 weeks reached the 25th centile for OFC later than those born at <34 weeks not exposed (median: 15 vs. 7 months; P = 0.004). Gestational age and maternal illicit drug use were strongly associated with growth, but the effect of combination therapy exposure was marginal (adjusted coefficients: weight, -0.10 [P = 0.019]; height, -0.12 [P = 0.008]; and OFC, -0.14 [P = 0.001]). Although the effect of combination therapy exposure is minimal, long-term monitoring of these children is important. PMID- 16249714 TI - Is sexual contact with sex workers important in driving the HIV epidemic among men in rural Zimbabwe? AB - OBJECTIVE: To establish the importance of commercial sex in driving the HIV epidemic in the general population by determining risk factors for HIV infection among male mine and farm workers and estimating the fraction of prevalent HIV infections attributable to sexual contact with sex workers (SWs). SETTING: Five commercial farms and 2 mines in Mashonaland West, Zimbabwe. METHODS: A cross sectional interviewer-administered questionnaire and urine survey of 1405 male workers. Urine samples were tested for HIV antibodies by a particle agglutination test and enzyme-linked immunosorbent assay and for Chlamydia trachomatis and Neisseria gonorrhoeae using a polymerase chain reaction assay. RESULTS: The overall prevalence of HIV antibodies was 27.3% (95% confidence interval [CI]: 24.8 to 29.5), that of C. trachomatis was 1.5% (95% CI: 1.0 to 2.1), and that of N. gonorrhoeae was 0.5% (95% CI: 0.1 to 0.9). A total of 48.4% (95% CI: 45.8 to 51.0) of men reported ever having had sexual contact with an SW, and 29.3% (95% CI: 26.9 to 31.7) reported contact in the past year. HIV was more common among men who reported SW contact on univariate (1.9% [95% CI: 1.5 to 2.4]) and multivariate (1.4% [95% CI: 1.0 to 1.8]) analysis after adjusting for confounding. HIV was also strongly associated with self-reported genital ulceration in the previous 6 months (adjusted odds ratio [OR] = 3.1, 95% CI: 2.2 to 4.3). Genital ulceration and SW contact were highly correlated. A total of 19.6% of HIV infections in men could be attributed to ever having had sexual contact with an SW (95% CI: 10.8 to 27.6). CONCLUSIONS: An appreciable proportion of HIV infection in men is attributable to sexual contact with SWs. Consideration should be given to developing interventions that target male clients of SWs. PMID- 16249715 TI - Safe discontinuation of primary pneumocystis prophylaxis in Southern Indian HIV infected patients on highly active antiretroviral therapy. PMID- 16249716 TI - Impact of coinfection with HIV-1 and GB virus C in patients receiving a ritonavir boosted HAART regimen: a substudy to the MaxCmin1 trial. PMID- 16249717 TI - Employment of off-duty staff: a strategy to meet the human resource needs for a large PMTCT program in Zambia. PMID- 16249718 TI - Application of denaturing gradient gel electrophoresis (DGGE) to the analysis of endodontic infections. AB - The recent expanding use of cultivation-independent techniques for bacterial identification is reliant on the lack of knowledge of the conditions under which most bacteria are growing in their natural habitat and the difficulty to develop culture media that accurately reproduce these conditions. A molecular method that has been recently used in several areas to examine the bacterial diversity living in diverse environments is the denaturing gradient gel electrophoresis (DGGE). In DGGE, polymerase chain reaction (PCR)-generated DNA fragments of the same length but with different base-pair sequences can be separated. Separation is based on electrophorectic mobility of a partially melted double-strand DNA molecule in polyacrylamide gels, which is decreased when compared with that of the completely helical form of the molecule. Molecules with different sequences may have a different melting behavior and will therefore stop migrating at different positions in the gel. Application of the PCR-DGGE method in endodontic research has revealed that there are significant differences in the predominant bacterial composition between asymptomatic and symptomatic cases. This suggests that the structure of the bacterial community can play a role in the development of symptoms. In addition, new bacterial phylotypes have been disclosed in primary endodontic infections. PCR-DGGE has also confirmed that intra-radicular infections are a common finding in root-filled teeth associated with persistent periradicular lesions. The microbiota in failed cases significantly vary from teeth to teeth, with a mean number of species far higher than previously shown by culturing approaches. Application of the PCR-DGGE technique in endodontic microbiology research has the potential to shed light on several aspects of the different types of endodontic infection as well as on the effects of treatment procedures with regard to infection control. PMID- 16249719 TI - The influence of different factors on the survival of root canal fillings: a 10 year retrospective study. AB - The aim of the present study was to determine the survival time of root canal fillings performed in 1990 and 1991 at the Dental School, Philipps University of Marburg, Germany. Data were collected retrospectively from the dental records and control radiographs were evaluated. The survival probability was determined by Kaplan-Meier analysis. Intergroup differences were verified with the log-rank test. The 914 evaluated root canal fillings yielded an overall 10-yr cumulative survival probability of 0.74. The factors baseline periapical condition (p < 0.001), length (p < 0.001), condensation (p < 0.001), vitality (p < 0.001), and pain symptoms (p = 0.005) were found to have a significant influence on the long term success of root canal fillings. Higher survival rates were recorded for teeth with healthy periapical conditions, root canal fillings of the correct length, homogeneously condensed root canal fillings, root canal fillings in previously vital teeth, and teeth that had been asymptomatic during treatment. A nonsignificant influence was recorded for the parameters operator (p = 0.606) and retreatment (p = 0.196). PMID- 16249720 TI - The effect of endodontic therapy on periapical exudate neutrophil elastase and prostaglandin-E2 levels. AB - This study aimed to investigate periapical exudate neutrophil elastase (NE) and prostaglandin E2 (PGE2) levels and their relationships with clinical symptoms, and to determine the changes of their levels following first treatment visit. Periapical exudate samples were collected from the canals of 34 nonvital single rooted teeth at two sequential treatment visits. Periapical exudate NE and PGE2 levels were found to be higher in the presence of clinical symptoms (pus discharge, swelling) (p < 0.05). The canals of teeth with larger periapical radiolucent area (>or=1 cm) contained more PGE2 levels than with smaller ones (<1 cm) (p < 0.05). Periapical exudate NE levels were significantly correlated with PGE2 levels (p < 0.05), and their levels at first treatment visit did not change following root canal therapy (p > 0.05). The periapical exudate NE and PGE2 levels may regulate periapical disease expression, but the results of this study were unable to reveal this association. PMID- 16249721 TI - A comparative study of three different root canal curvature measurement techniques and measuring the canal access angle in curved canals. AB - In the first part of this study the Schneider (S), Weine (W), and Long-Axis (LA) techniques are used for comparing the measurement of canal curvature. One hundred mandibular first and second molar teeth were selected. Radiographs were taken after inserting size 10 K-files into the mesiobuccal root canals. The radiographic findings were digitized on a computer, and the three different curvature angles were measured from drawings of the same root canal and compared statistically. ANOVA showed that there were significant differences between the curvature angle values determined using each technique (p < 0.001). In the second part of this study the term "canal access angle" (CAA) was introduced and it was defined by examining the morphology of canal curvature. Canal length, curvature distance (y), curvature height (x), Schneider angle, and the newly defined CAA were evaluated statistically. Using a multiple regression analysis, the CAA was significantly related to x (p < 0.001) and y (p < 0.005). There was a positive correlation (r = 0.74) between the CAA and curvature height (x). The results indicated that the CAA is a more effective way of evaluating the root canal curvature. PMID- 16249723 TI - Fibroblast growth factor-2 stimulates hyaluronan production by human dental pulp cells. AB - Hyaluronan (HA), is a high molecular mass extracellular matrix constituting connective tissue and plays a critical role in not only homeostasis but also inflammatory and wound-healing responses. In this study, we investigated the effect of fibroblast growth factor (FGF)-2 on the production of HA by human dental pulp cells (HDPC). An inhibition binding-protein assay showed that FGF-2 increased HA production by HDPC. In addition, expression of mRNA of hyaluronan synthase (HAS) 1 and HAS 2, both of which are related to the production of high molecular mass of HA, but not HAS 3, was enhanced in FGF-2-stimulated HDPC. These results provide new evidence for the involvement of FGF-2 in the regulation of HA production by HDPC possibly through HAS 1 and HAS 2. PMID- 16249722 TI - A real time quantitative PCR analysis and correlation of COX-1 and COX-2 enzymes in inflamed dental pulps following administration of three different NSAIDs. AB - Dental pain is encountered daily by clinicians. Nonsteroidal anti-inflammatory drugs (NSAIDs) commonly used for pain management are traditionally cyclooxygenase 1 (COX-1) and cyclooxygenase-2 (COX-2) inhibitors, and more recently selective COX-2 inhibitors. This study was designed to identify and quantify COX-1 and COX 2 gene expression level in inflamed rat molar pulps after administration of three NSAIDs: Celebrex, Vioxx, and Advil. Fifty male Wistar rats had their first and second molar pulps exposed and sealed with Cavit for 4 days. Rats were randomly divided into the three drug groups and two control groups. RNA was isolated from the rat pulps. Real Time Quantitative Reverse Transcriptase-Polymerase Chain Reaction assay, a relatively new PCR technique, was used to quantify COX-1 and COX-2 mRNA. Statistical analysis demonstrated no significant differences in COX-1 and COX-2 levels among the drug groups. However, Vioxx and Advil significantly reduced COX-2 expression levels compared to inflamed (positive control) pulps (p < 0.05). PMID- 16249724 TI - Interfacial strength of Resilon and gutta-percha to intraradicular dentin. AB - Strengthening of Resilon-filled roots via an adhesive interface should be reflected by improvement in the interfacial strength and dislocation resistance between the root fillings and intraradicular dentin. This study compared the interfacial strengths of Resilon/Epiphany and gutta-percha/AH Plus using a thin slice push-out test design. Failure modes of root slices after push-out testing were examined with environmental scanning electron microscopy. The gutta-percha group exhibited significantly higher interfacial strength than the Resilon group, when premature failures that occurred in Resilon root slices were included in the statistical analysis. The gutta-percha root slices failed exclusively along the gutta-percha/sealer interface. The Resilon root slices failed predominantly along the sealer/dentin interface with recognizable, fractured resin tags. Detachment of the Resilon from the Epiphany sealer was also surprisingly observed in some specimens. The similarly low interfacial strengths achieved with both types of root filling challenges the concept of strengthening root-filled teeth with the new endodontic material. PMID- 16249725 TI - An in vitro assessment of iodoform gutta-percha. AB - The purpose of this study was to test the ability of a commercially available iodoform gutta-percha, to delay infiltration of Enterococcus faecalis using a microleakage model. Seventy extracted single-rooted teeth were decoronated and biomechanically prepared using hand and rotary instruments. Thirty roots were obturated laterally with iodoform gutta-percha and another 30 with regular gutta percha. Both groups were suspended in sterile BHI broth. An inoculum of E. faecalis suspension was placed at the coronal end of each root, incubated and replenished daily. The apical broth was observed for turbidity, indicating bacterial microleakage. Samples were observed for 32 days, and data was analyzed to compare microleakage between the two groups. The results showed no significant difference between the iodoform and regular gutta-percha samples in delaying microleakage of E. faecalis (p > 0.05). PMID- 16249727 TI - Erbium:YAG laser versus ultrasonic in preparation of root-end cavities. AB - The aim of this in vitro study was to compare the sealing effectiveness of the Erbium:YAG laser to the sealing effectiveness of an ultrasonic device in the preparation of similar retrograde cavities using different retrograde filling materials. After root-canal instrumentation and filling, apices of 60 single rooted teeth were resected. Retrograde class I cavities 3 mm deep were prepared using an ultrasonic device (group A) and Er:YAG laser (group B). An ultrasonic unit was used with CT-5 retrotip at the frequency of 32 KHz. Laser beam parameters were a pulse of very short duration (100 micros), energy of 280 mJ, and repetition rate of 10 Hz. Cavities of each group of 10 samples were filled with mineral trioxide aggregate (MTA), Super-EBA, and IRM. Microleakage was measured using a fluid transport model. The results showed that cavities prepared with Er:YAG laser have significantly lower microleakage for all tested materials. PMID- 16249726 TI - Chelation in root canal therapy reconsidered. AB - The aim of this study was to assess interactions of EDTA and citric acid (CA) with sodium hypochlorite (NaOCl), the indispensable endodontic irrigant. Other chelators were simultaneously evaluated as possible alternatives: sodium triphosphate (STP), amino tris methylenephosphonic acid (ATMA), and 1- hydroxyethylidene-1, 1-bisphosphonate (HEBP). Available chlorine was titrated in chelator-NaOCl solutions. All chelators other than HEBP and STP caused an almost complete, immediate loss of available chlorine in solution. Atomic absorbtion spectrometry and SEM evaluation of root canal walls of instrumented teeth indicated that NaOCl had no negative effect on calcium-complexing ability of chelators. STP was too weak a complexing agent to warrant further studies. Finally, CA-, EDTA-, and HEBP-NaOCl mixtures were evaluated for their antimicrobial capacity. Again, EDTA and CA negatively interfered with NaOCl, while HEBP did not. PMID- 16249728 TI - Comparison of the cutting efficiency of two ultrasonic units utilizing two different tips at two different power settings. AB - The objective of this in vitro study was to set up a testing protocol and utilize it to evaluate and compare the cutting efficiency of ultrasonic units. Evaluation of two ultrasonic units utilizing two different tips was done by measuring the weght of a dentin specimen before and after ultrasonic treatment for set time periods. The difference in weight was the amount of dentin removed. The ultrasonic units tested were the P5 Booster (Staelec, France) and the Spartan (Obtura-Spartan, Fenton, MO) machines. The tips tested were the CPR-2D and ETD20 types. The units were tested at maximum and medium power. All the experimental variables, ultrasonic unit type, power setting and tip type were found to have an effect on the cutting efficiency (P<0.0001). The P5 ultrasonic unit was more effective at dentin removal than the Spantan ultrasonic unit at both power settings. The ETD-20 tip was more effective at dentin removal than the CPR-2D tip on both units at both power settings. PMID- 16249729 TI - An objective evaluation comparing the physical properties of two brands of stainless steel endodontic hand files. AB - The purpose of this study was to evaluate the physical properties of a flexible stainless steel K-type hand file recently introduced by Brasseler USA (F-Style files; Brasseler USA, Savannah, GA), in comparison to those of a well-studied instrument (Flex-o-files; Maillefer-Dentsply, Ballaigues, Switzerland), that has been on the market for more than 20 yr. The physical properties measured included torque at failure, angular deflection at failure, flexibility, and consistency of diameter at 3 mm from the cutting tip; and the evaluations were carried out on size #10 through #50 files of each of the two brands being tested. The results indicated that the Brasseler instruments were inherently more flexible, but had smaller diameters, lower torque, and angular deflection values at failure, than those of the Maillefer files. PMID- 16249730 TI - Endodontic implications of bisphosphonate-associated osteonecrosis of the jaws: a report of three cases. AB - Bisphosphonates are commonly used in medicine to maintain bone density in patients with certain nonneoplastic diseases or cancers. A serious adverse effect of bisphosphonates that has substantial dental significance is osteonecrosis that appears to uniquely affect the mandible and maxilla without occurring in other bones of the skeleton. Patients with bisphosphonate associated osteonecrosis of the jaws may present with pain and exposed necrotic bone. This has substantial clinical implications because surgical procedures (including extractions or endodontic surgical procedures) are contraindicated in the jaws of these patients and the presenting pain may mimic pain of odontogenic origin. This report describes three patients with bisphosphonate associated osteonecrosis and emphasizes the endodontic implications of managing these patients. PMID- 16249733 TI - Routine intraoperative stenting for renal transplant recipients. AB - BACKGROUND: Major urological complications (MUCs) after kidney transplantation contribute to patient morbidity and compromise graft function. Ureteric stents have been successfully used to treat such complications and a number of centers have adopted a policy of universal prophylactic stenting, at the time of graft implantation, to reduce the incidence of urine leaks and ureteric stenosis. METHODS: In conjunction with the Cochrane Renal Group we searched the Cochrane Central Register of Controlled Trials, MEDLINE, EMBASE, reference lists of articles, books and abstracts and contacted companies, authors and experts to identify randomized controlled trials examining the use of stents in renal transplantation. The primary outcome was the incidence of MUCs and data on this statistic was pooled and analyzed using a random effects model. RESULTS: Seven randomized controlled trials (1154 patients) of low or moderate quality were identified. The incidence of MUCs was significantly reduced (relative risk [RR] 0.24; 95% CI: 0.07 - 0.77; P=0.02; number needed to treat = 13) by prophylactic ureteric stenting. Urinary tract infections were more common in stented patients (RR 1.49), unless the patients were prescribed 480 mg cotrimoxazole once daily. With this antibiotic regime the incidence of infection was equivalent between the two groups (RR 0.97). Stents appeared generally well tolerated, although trials using longer stents (> or = 20 cm) for longer periods of time (>6 weeks) reported more problems with encrustation and migration. CONCLUSIONS: Universal prophylactic stenting reduces the incidence of MUCs and should be recommended on the basis of currently available randomized controlled trials. PMID- 16249734 TI - Maintenance immunosuppression with target-of-rapamycin inhibitors is associated with a reduced incidence of de novo malignancies. AB - BACKGROUND: Immunosuppressive drug therapy has been identified as one etiological factor in the increased incidence of and deaths from malignancies in renal transplant recipients. In animal models, calcineurin inhibitors have a positive growth effect, whereas target-of-rapamycin (TOR) inhibitors have a negative growth effect on malignant cells. METHODS: A multivariate analysis of posttransplant malignancies in 33,249 deceased donor primary solitary renal recipients reported by 264 kidney transplant programs to the Organ Procurement and Transplantation Network database from July 1, 1996 to December 31, 2001 was performed. Data were censored at 963 days to allow comparable follow-up time among drug treatment groups. The incidence and relative risks of any de novo malignancy (skin and solid) and for non-skin solid malignancies in patients receiving TOR inhibitors compared to patients receiving calcineurin inhibitors were the primary endpoints. RESULTS: The incidence rates of patients with any de novo posttransplant malignancy were 0.60% with sirolimus/everolimus alone, 0.60% with sirolimus/everolimus + cyclosporine/tacrolimus, and 1.81% with cyclosporine/tacrolimus (P<0.0001); the rates with a de novo solid tumor were 0%, 0.47%, and 1.00%, respectively. In the Cox regression model the relative risk associated with sirolimus/everolimus immunosuppression for any de novo cancer was 0.39 (95% CI: 0.24-0.64; P=0.0002) and for de novo solid cancer was 0.44 (0.24 0.82; P=0.0092). Other significant risk factors were male sex, adult age group, white race, and history of a malignancy. CONCLUSIONS: Maintenance immunosuppression with the TOR inhibitor drugs, sirolimus and everolimus, is associated with a significantly reduced risk of developing any posttransplant de novo malignancy and non-skin solid malignancy. PMID- 16249735 TI - Ethnic differences in donation-related characteristics among potential hematopoietic stem cell donors. AB - BACKGROUND: Although the National Marrow Donor Program has been highly successful at recruiting ethnic minorities as potential hematopoietic stem cell donors, there have been no systematic investigations of whether donor characteristics that might be linked to the donation experience vary by ethnicity. METHODS: Questionnaires assessing four domains-demographic, volunteer-related, general psychosocial, and donation-related-were mailed to potential donors after they were contacted as a preliminary match for a patient and had agreed to donate. In all, 1,679 potential donors completed and returned a predonation questionnaire. Data from potential donors belonging to five major ethnic groups were analyzed; white, black, Asian/Pacific Islander, Hispanic, and Native American. RESULTS: Bivariate analyses indicated that virtually all factors in the four domains were associated with ethnicity. Direct discriminant function analysis identified three significant functions. The most striking of the three functions indicated that Asian Americans were more highly educated, more ambivalent (reluctant about donation), more concerned (medical, work/family), and more anxious and depressed than all other ethnic groups. Key differences among other ethnic group members were also identified. CONCLUSIONS: This study provides the first evidence of ethnic group differences in key predonation variables. Findings suggest that Asian/Pacific Islanders possess a number of characteristics that are known psychosocial risk factors for less positive postdonation outcomes and that more intensive pre and postdonation contact with this group may be necessary. Strategies for improving future research in this area are discussed. PMID- 16249736 TI - Liver transplantation for hepatocellular carcinoma in cirrhosis within the Eurotransplant area: an additional option with "livers that nobody wants". AB - BACKGROUND: Liver transplantation is recognized as the treatment of choice for small hepatocellular carcinomas (HCC) in patients with end-stage liver failure. However, because of limited organ availability, not all those who qualify can benefit from it. METHODS: Over a 3-year period, we accepted and subsequently transplanted 10 deceased donor liver allografts allocated through Eurotransplant. These organs had been officially offered to and rejected by other transplant centers a total of 40 times due to medical or logistical reasons prior to our acceptance. They were implanted into patients in the waiting list with HCC and cirrhosis. Recipients without HCC transplanted with such "undesirable" grafts were not included in this study. RESULTS: Two patients had initial poor graft function but subsequently recovered. There was one arterial complication requiring reintervention. Median intensive care unit and hospital stays were 6 and 28 days respectively. One patient developed renal insufficiency, but recovered after 3 months. One patient developed HCC recurrence in the allograft and underwent a successful atypical liver resection 23 months after transplantation. All patients are currently alive, with follow-up periods ranging from 5 to 36 months. CONCLUSIONS: Liver transplantation with such "livers that nobody wants" constitutes an additional option for patients with HCC and cirrhosis. The risk-benefit ratio in these instances should be evaluated on a case-by-case basis. PMID- 16249737 TI - Predictive factors of anemia within the first year post renal transplant. AB - BACKGROUND: The aim of our study was to identify the independent factors that might predict anemia at 6 (M6) and 12 (M12) months posttransplantation. METHODS: Postrenal transplant anemia (PTA) was defined as having a hemoglobin (Hb) level below 13 g/dl for men and below 12 g/dL for women. In this study, we included all the recipients who received a renal transplant in 2001 at our department, and for whom the graft was still functioning 1 year later (n=92). RESULTS: Anemia was observed in 78%, 35.5% and 25% of patients at day (D)0 and at M6 and M12, respectively. Iron deficiency was found in 14% of patients at D0 and in 13% of patients at M12. A total of 59.8% of patients had received at least one blood transfusion in the postoperative period, whereas 41.3% of patients had received recombinant erythropoietin (rEpo) therapy within the first months posttransplantation. In multivariate analysis, the independent predictive factors of anemia at M6 were Epo level at D0, initial nephropathy (polycystic kidney disease vs. others), posttransplantation rEpo therapy, hematocrit at M3, platelets at D7, and sirolimus therapy. The independent predictive factors of anemia at M12 were Epo level at D0, platelets at D7, delayed graft function (DGF), creatinine clearance at M12, serum creatinine at M12, and Hb level at M6. CONCLUSIONS: The prevalence of PTA was 25% at M12. DGF, renal function at M12, and anemia at M6 were independent risk factors for still having anemia at M12. PMID- 16249739 TI - Race/ethnicity, poverty status, and renal transplant outcomes. AB - BACKGROUND: There are known racial disparities in renal graft survival. Data are lacking comparing associations of race/ethnicity and socioeconomic status with graft failure and functional status after transplantation. Our goal was to test if African-American and Hispanic race/ethnicity and poverty are associated with worse outcomes following renal transplantation. METHODS: We performed a retrospective cohort study using a nationwide registry (United Network for Organ Sharing). We studied 4,471 adults who received renal transplants in 1990. Outcomes were graft failure and functional status over 10 years. RESULTS: Cumulative incidence of graft failure was higher among African-Americans and Hispanics than whites (77% vs. 64% vs. 60 %; P<0.001) and among transplant recipients living in the poorest areas (70% vs. 58% in the richest; P<0.001). African-American and Hispanic race/ethnicity were independently predictive of graft failure (RR 1.8, 95% CI 1.6-1.9; RR 1.3, 95% CI 1.2-1.6, respectively) in multivariate analyses but poverty status was not (RR 1.0, 95% CI 0.9-1.1). Days with impaired functional status were higher for African-Americans compared to whites (RR 1.6, 95% CI 1.3-1.9) but not independent of poverty. Poverty was independently associated with impaired functional status (RR 1.3, 95% CI 1.0 1.6). CONCLUSIONS: African-Americans and Hispanics had higher rates of graft failure compared to whites after adjustment for poverty and other covariates whereas poverty, but not race/ethnicity, was related to functional status following renal transplantation. National datasets should include individual level measures of socioeconomic status in order to improve evaluation of social and environmental causes of disparities in renal transplant outcomes. PMID- 16249738 TI - A multicenter, prospective study of C2-monitored cyclosporine microemulsion in a U.S. population of de novo renal transplant recipients. AB - BACKGROUND: Monitoring cyclosporine microemulsion (CsA-ME; Neoral) exposure 2 hours postdose (C2) has been reported to optimize the efficacy and safety of CsA ME therapy. The addition of induction therapy to a maintenance regimen including CsA-ME C2 monitoring has not been evaluated. METHODS: In all, 123 adult renal transplant recipients were recruited at 14 U.S. centers for this 6-month study. CsA-ME dose was to be titrated to attain C2 targets of 1700 and 1500 ng/ml during posttransplant months 1 and 2, respectively. After 2 months, patients were randomized to one of two groups with different, decreasing C2 targets. Basiliximab, mycophenolate mofetil, and corticosteroids completed the study immunosuppression. RESULTS: Of the 119 evaluable patients, 76% were male, 22% African American, and 66% deceased donor recipients. Biopsy-proven acute rejection occurred in 10 patients (9.3%); there were two failed grafts and one death. Serum creatinine and calculated GFR values suggest good renal function, with month 6 medians of 1.5 ng/ml and 67 ml/min/1.73 m. Safety and tolerability assessments revealed no unexpected outcomes. Observed C2 levels were generally lower than protocol targets, particularly in the first weeks posttransplantation. CONCLUSIONS: The striking efficacy and outcomes may have been achieved in this study with lower C2 levels of CsA-ME because of the addition of basiliximab induction. PMID- 16249740 TI - Improving the prediction of donor kidney quality: deceased donor score and resistive indices. AB - BACKGROUND: The deceased donor score (DDS), expanded criteria donor (ECD) definition, and resistive index (RI) were developed for pretransplant evaluation of donors. DDS and ECD are determined by a calculation of risk from donor variables, while RI is determined from flow characteristics of kidneys during machine preservation (MP). This study was designed to compare DDS, ECD status, and RI as predictors of outcome after deceased donor transplantation. We were also interested to see if DDS or ECD could identify kidneys most likely to benefit from MP. METHODS: We retrospectively reviewed 48,952 deceased donor renal transplants reported to UNOS from 1997-2002. DDS (0-39 pts.), ECD status (+ or ), and preservation technique (MP vs. cold storage [CS]) were determined in all cases. RI during MP was studied in a single-center cohort of 425 transplants. RESULTS: DDS was superior to ECD status and RI in its correlation with early and late renal function after transplantation. DDS identified a subgroup of ECD- kidneys, those with DDS > or = 20 pts, that functioned significantly below expectation and similar to ECD+ kidneys. Benefits of MP, which include improved early graft function and a trend towards longer graft survival, were greatest in the group of kidneys with DDS > or = 20 pts. CONCLUSIONS: DDS was the best predictor of outcome after deceased donor renal transplantation and may be useful in identifying kidneys most likely to benefit from MP. PMID- 16249741 TI - New prognostic scoring model for liver transplantation in patients with non acetaminophen-related fulminant hepatic failure. AB - BACKGROUND: Many patients with fulminant hepatic failure die before receiving liver transplantation because of the difficulty of pinpointing the suitable timing for liver transplantation. The revised King's College criteria are useful for patients with acetaminophen-related fulminant hepatic failure; however, in those with non-acetaminophen-related fulminant hepatic failure, a new prognostic system that can accurately identify the suitable timing for liver transplantation is required. METHODS: Using the first sample consisted of eighty patients with fulminant hepatic failure, we examined 2-week poor prognostic parameters at the time of diagnosis of fulminant hepatic failure (day 1) and on days 4, 8, and 15, respectively, and a 2-week prognostic scoring model was constructed. To confirm the accuracy of this model, validation was performed in the second sample consisting of 26 patients. RESULTS: Cause of fulminant hepatic failure (hepatitis B virus or indeterminate), hepatic coma grade (III or IV), systemic inflammatory response syndrome (yes) and ratio of total to direct bilirubin (> 2.0) were associated with 2-week outcomes during days 1-15. Each of these four parameters was valued at +1. The 2-week survival rate in patients scoring <3 was > or = 80% in contrast to less than 30% in patients scoring > or = 3. When this scoring model was applied to the second sample, the sensitivity, specificity, and positive and negative predictive values were 87.5%, 90.0%, 93.3%, and 81.8%, respectively. CONCLUSIONS: This scoring model may be useful for predicting 2-week outcomes and determining the suitable timing for liver transplantation in patients with non-acetaminophen-related fulminant hepatic failure. PMID- 16249742 TI - Free fatty acids are associated with obesity, insulin resistance, and atherosclerosis in renal transplant recipients. AB - BACKGROUND: Insulin resistance (IR) may be implicated in the pathogenesis of atherosclerosis in renal transplant recipients (RTRs) and be contributed to, in part, by free fatty acids (FFAs), produced in excess in centrally obese individuals. The aim of this study was to determine the prevalence of IR and the relationships between FFAs, central obesity, and atherosclerosis in a cohort of prevalent RTRs. METHODS: Observational data were collected on 85 RTRs (mean age 54 years; 49% male, 87% Caucasian). Fasting serum was analyzed for FFAs, glucose, and insulin; IR was calculated using the homeostasis model assessment (HOMA-IR) score. Vascular structure was assessed by carotid intima-media thickness (IMT) measurement. Linear regression analyses were performed to determine the factors associated with IR and atherosclerosis. RESULTS: IR occurred in 75% of RTRs, and FFA levels were independently associated with its occurrence (beta: -0.55, 95% CI: -1.02 to -0.07, P = 0.02). Other variables independently associated with IR were male sex, body mass index, central obesity, diabetes, systolic blood pressure and corticosteroid use. There was a significant correlation between FFA levels and IMT (r = 0.3, P=0.01). On multivariate analysis, IMT correlated with elevated FFA (beta: 0.07, 95% CI: 0.02-0.12, P = 0.007), diabetes mellitus (P = 0.05), older age (P < 0.002), and a body mass index >25 kg/m (P = 0.002). CONCLUSIONS: FFAs are associated with the development of IR and may be involved in the pathogenesis of atherosclerosis in RTRs. Additional studies are required to explore these associations further before considering whether an interventional trial aimed at lowering FFA would be a worthwhile undertaking. PMID- 16249743 TI - New-onset diabetes after kidney transplantation: an application of 2003 International Guidelines. AB - BACKGROUND: The 2003 International Consensus Guidelines defined new-onset diabetes after transplantation. This study determined the risk of new-onset diabetes following kidney transplantation using these criteria. METHODS: Consecutive nondiabetic patients who received kidney transplantation between August 2001 and March 2003 (recent, n=61) and before August 2001 (earlier, n=61) were retrospectively evaluated. RESULTS: In all, 74% in the recent group and 56% in the earlier group developed diabetes by 1 year posttransplant. Median time to diabetes development was 23 days in the recent vs. 134 days in the earlier group (P=0.0304). Most patients developed diabetes within 60 days after transplantation. Immunosuppression was the strongest correlate of diabetes development; tacrolimus and cyclosporine A treatments were associated with increased risk. The rate of development was also greater when rapamycin was added to tacrolimus, compared to when it was not. The risk was double in African Americans compared to whites. Age, body mass index, family history of diabetes, and etiology of renal failure did not predict diabetes; however, the mean age of patients was greater than previously reported. CONCLUSIONS: The majority of patients are at risk of developing new-onset diabetes within a short time after kidney transplantation. The risk may be due to preexisting risk factors, immunosuppressive agents, or older age. The significance of these findings is not clear, but demands appropriate follow-up studies related to glycemia, end-organ complications, and graft function. It remains to be determined whether the 2003 International Consensus Guidelines are adequate to appropriately diagnose diabetes in the posttransplant time period, with special emphasis on the first 3 months. PMID- 16249744 TI - Preferential allocation of marginal kidney allografts to elderly recipients combined with modified immunosuppression gives good results. AB - BACKGROUND: There is an increasing tendency to allocate kidneys from marginal donors in older recipients. This combination optimizes the uses of an expanded donor pool but demands attention for the higher nephrotoxic sensitivity of the kidney and the increased immunosuppression vulnerability of the elderly recipients. We aimed to reduce these hazards by means of a calcineurin-free induction therapy followed by a maintenance regimen targeted to minimize/withdraw steroid. METHODS: Eighty-eight single (43%) or double (57%) transplant recipients (58.4+/-5.7 years) from 88 marginal donors (67+/-8.3 years) received monoclonal anti-IL-2 receptor antibodies, mycophenolate mofetil (MMF), and steroid. When serum creatinine was less than 2.6 mg/dL, tacrolimus was started and MMF was withdrawn when the tacrolimus trough level was above 15 ng/ml. Steroid was tapered to 5 mg at day 45 and then progressively reduced. RESULTS: Overall patient and graft survival at the first and fourth year were respectively 100 and 96%, and 98 and 79%. Acute rejection rate was 13.6% (12/88), creatinine clearance remained stable (48.2 ml/min at the sixth month, 50.9 ml/min at 48th month). At the first, second, third, and fourth years, 23, 69, 80, and 100% of recipients were off steroids. For those on steroids, mean dose was respectively 2.6 mg/day from month 12. No recipient re-assumed steroids CONCLUSIONS: In the "old-for-old" allocation, the calcineurin-inhibitor avoidance at induction and the steroid withdrawal/minimization during the tacrolimus-based maintenance regimen allow a low acute rejection rate, a stable renal function, and favorable recipient and graft outcomes. PMID- 16249745 TI - Immediate extubation of children following liver transplantation is safe and may be beneficial. AB - BACKGROUND: Immediate tracheal extubation of selected adult patients after orthotopic liver transplant (OLT) is common practice. We hypothesized that selected children may be safely extubated immediately after OLT and avoid potentially deleterious effects of artificial ventilation and sedation. METHODS: After June 2002, we chose immediate extubation unless a specific contraindication was identified. Charts of all children undergoing OLT between June 2002 and February 2005 were reviewed to audit safety and outcome of this approach. Comparative data were obtained for children undergoing first elective OLT at other UK centers. RESULTS: Forty-six cadaveric liver transplants were performed in 40 patients: 26 of 34 (76%) elective transplants and 4 of 12 (33%) urgent transplants were extubated immediately after surgery. Eight of 14 (57%) children weighing less than 10 kg were successfully extubated. One child required reintubation after developing transfusion-related acute lung injury. There were no other events compromising patient or graft. Small recipient size, split/reduced grafts, preexisting respiratory disease, retransplantation, and acute liver failure did not individually preclude successful immediate extubation. After elective OLT, the mean duration of intensive care stay was significantly shorter in the extubated group than in those who were ventilated (2.5 vs. 6.1 days, P<0.01). All children receiving a liver transplant at other UK centers in 2003 were ventilated postoperatively. However, the median duration of intensive care stay (2 days) was the same as in our series. CONCLUSIONS: Immediate extubation of selected children after OLT is safe. It may enhance patient recovery, benefit graft physiology, and reduce intensive care requirement. PMID- 16249746 TI - Impact of graft hepatic vein inferior vena cava reconstruction with graft venoplasty and inferior vena cava cavoplasty in living donor adult liver transplantation using a left lobe graft. AB - BACKGROUND: Hepatic venous reconstruction is critical in living donor adult liver transplantation (LDALT) because outflow obstruction in small for size graft may lead to graft dysfunction or loss. We describe the usefulness of venoplasties of the graft hepatic vein (HV) and graft HV-recipient inferior vena cava (IVC) reconstruction in LDALT using a left lobe graft. METHODS: Sixty patients who underwent LDALT were studied. We divided the patients into following two groups: venoplasty group (n=30) and control group (n=30). For the patients with venoplasty group, venoplasty of the graft and recipient IVC cavoplasty was made to widen the orifice. Comparison examination of a background factors and postoperative bilirubin and the ascites was carried out. RESULTS: The mean graft volume standard liver volume ratio (GV/SLV) did not have the difference at 41.7% of venoplasty group, and 42.1% of control group (p=NS). The diameter of the hepatic vein in control and venoplasty group before and after venoplasty is 26.9+/-5.5, 28.2+/-2.9, and 34.1+/-3.9 mm, respectively. The diameter of the hepatic vein after venoplasty is larger than that of before venoplasty and of control (P<0.05). Mean total bilirubin level on postoperative day (POD) 7 is 13.8+/-9.3 mg/dl in control group and 7.0+/-3.3 mg/dl in venoplasty group (P<0.05). Mean amount of ascites on POD 7 and 14 are 1576+/-1113 and 1397+/-1661 cc in control group, and 736+/-416 and 550+/-385 cc in venoplasty group, respectively (P<0.05). Two-year survival rate is 75.2 % in control group and 86.6 % in venoplasty group (P<0.05). CONCLUSIONS: We conclude that in LDALT using left lobe graft, HV-IVC reconstruction with graft venoplasty and IVC cavoplasty is useful not only to prevent outflow block but also to improve graft function. PMID- 16249747 TI - Nonmyeloablative conditioning does not prevent telomere shortening after allogeneic stem cell transplantation. AB - BACKGROUND: Stem cell transplantation (SCT) may be associated with premature aging of the hematopoietic stem cells. Telomere length reflects the proliferative history of a cell. In most studies published so far on telomere dynamics after myeloablative allogeneic SCT, recipients had shorter telomeres than their respective donors, thus reflecting "accelerated aging" of hematopoietic cells. We evaluated telomere dynamics in patients who underwent transplantation with nonmyeloablative protocols, assuming that the decreased intensity of chemotherapy might prevent telomere attrition. METHODS: Telomere length was measured using FISH-FACS method. Telomeres of recipients were compared to their respective donors. Twenty-three consecutive patients after nonmyeloablative SCT were evaluated. A control group consisted of 10 donor-recipient pairs after conventional myeloablative transplantation. RESULTS: There was significant telomere shortening in both recipients of nonmyeloablative and myeloablative conditioning (0.487+/-0.65 kb, P=0.003; 0.361+/-0.50 kb, P=0.047 respectively). The extent of telomere shortening in the two groups was not different (P=0.64). There was no correlation between the degree of shortening and parameters such as time interval from transplant, age of donor or recipient, and the number of infused cells. CONCLUSIONS: This is the first study on telomere dynamics after nonmyeloablative conditioning SCT. The study demonstrates significant shortening of telomeres in recipients in spite of decreased intensity conditioning. Results of this study suggest that the main mechanism following transplantation is the proliferative stress imposed upon the stem cells and not direct damage by cytotoxic drugs. The different kinetics of restoration of hematopoiesis and the probable ongoing process of graft-versus-leukemia in the bone marrow do not prevent the attrition of telomeric ends of chromosomes. PMID- 16249748 TI - Sirolimus and tacrolimus trough concentrations and dose requirements after kidney transplantation in relation to CYP3A5 and MDR1 polymorphisms and steroids. AB - BACKGROUND: CYP3A5 and MDR1 polymorphisms have been shown to influence tacrolimus blood concentrations and dose requirements. The aim is to determine whether these polymorphisms also affect sirolimus trough concentrations and dose requirements after kidney transplantation. METHODS: Eighty-five renal transplant recipients receiving sirolimus were included. Twenty-four were treated with a combined sirolimus-tacrolimus regimen. Eighty-one patients received steroids. Sirolimus and tacrolimus were adjusted to a target therapeutic window. CYP3A5 (intron 3) and MDR1 (exons 12, 21, 26) genotypes were correlated to the adjusted trough concentrations and dose requirements for both sirolimus and tacrolimus. RESULTS: There were no significant correlation between adjusted sirolimus trough concentrations or dose requirements and genetic polymorphisms. In a multiple regression model, adjusted-prednisone dose was involved with a positive or negative effect when considering sirolimus dose requirements or adjusted concentrations, respectively. In the subgroup of patients treated by tacrolimus and sirolimus, adjusted tacrolimus doses were higher in patients carrying at least one CYP3A5 *1 allele (median 0.083 vs. 0.035 mg/kg for CYP3A5*3/*3 patients, P<0.05). Adjusted-prednisolone dose and CYP3A5 polymorphism explained up to 61% of the variability in tacrolimus dose requirements. CONCLUSIONS: Unlike tacrolimus, sirolimus adjusted trough concentrations and dose requirements seem not affected by CYP3A5 and MDR1 polymorphisms. Adjusted-prednisone dose has a significant impact on tacrolimus and sirolimus dose requirements. PMID- 16249750 TI - Influence of age in renal transplant infections: cases and controls study. AB - Kidney transplantation in elderly patients is a good therapeutic option, but the incidence of infections compared to younger patients must be studied. Case and control study was performed with 40 cases (patients older than 65) and 40 controls (younger than 65) receiving a kidney transplant between January 2000 and August 2002. In 32 cases (80%) and in 14 controls (32%), some type of infection appeared during the follow-up (odds ratio [OR] 5; 95% CI 1.6-20). The percentage of patients with bacterial infections was higher in the cases (70% vs. 28%; OR 5.7; 95% CI 1.9-20), especially for urinary infections. No differences for viral and fungal infections were observed in the two groups. Mortality rate was 13% in the cases (5% due to infections), whereas there was no controls' mortality. Although the number of bacterial infections was higher, kidney transplantation in elderly patients is a secure procedure. PMID- 16249749 TI - Lack of correlation between results of ABO-incompatible living kidney transplantation and anti-ABO blood type antibody titers under our current immunosuppression. AB - In this study, we examined the impact of preoperative anti-A/B antibody titers on the results of ABO-incompatible living kidney transplantation (LKT). In all, 167 recipients underwent ABO-incompatible LKT at our institution between 1989 and 2002. These patients were subdivided into those transplanted under cyclosporine with azathioprine or mizoribine (Group 1, n=78) and those transplanted under tacrolimus or mycophenolate mofetil (Group 2, n=89). Overall patient survival at 5 and 10 years was 93.8% and 88.0%, respectively. Overall graft survival at 5 and 10 years was 76.9% and 55.9%, respectively. Graft survival in the patients with anti-A/B IgG titers over 1:128 was significantly lower in group 1, whereas no significant correlation between the anti-A/B IgG titers and graft survival was found in group 2. In conclusion, no correlation between anti-A/B antibody titers and the results of ABO-incompatible LKT was seen after tacrolimus or mycophenolate mofetil application. PMID- 16249751 TI - Fulminant hepatic failure from primary hepatic lymphoma: successful treatment with orthotopic liver transplantation and chemotherapy. AB - Systemic lymphomas may involve the liver but rarely cause fulminant hepatic failure (FHF). Acute liver failure from primary hepatic lymphoma (PHL) is even less common with most patients succumbing to the sequelae of FHF before the correct diagnosis is made. We report a patient who underwent successful orthotopic liver transplant (OLT) and chemotherapy for FHF secondary to PHL. This previously-well male developed profound coagulopathy and encephalopathy 6 weeks after the onset of jaundice and fatigue. Workup failed to reveal the underlying cause of his liver failure and the patient soon required urgent OLT. Pathologic evaluation of his explanted liver revealed a malignant T-cell rich, large B-cell non-Hodgkin's lymphoma with widespread hepatocellular necrosis. The patient made an excellent clinical recovery and is undergoing CHOP-Rituxan chemotherapy. This scenario demonstrates that lymphoma should be considered in the differential diagnosis of FHF without clear etiology because of the potential for intervention with transplant and chemotherapy. PMID- 16249752 TI - Stronglyoides stercoralis infection from pancreas allograft: case report. PMID- 16249753 TI - Upregulation of HSP-72 in a human renal allograft during reflow. PMID- 16249755 TI - [IgE and respiratory disease]. AB - INTRODUCTION: IgE is known to provide the biological basis for allergy and immediate hypersensitivity. However, recent data provide some evidence that IgE responses are involved in other inflammatory processes apart from allergy, including several respiratory diseases. STATE OF THE ART: IgE binds to mast cells and basophils but also to other inflammatory cells, which are involved in non allergic processes. IgE has a role in antigen presentation and is implicated in a number of other immune mechanisms. In the airways, IgE plays an important role in bronchial hyperactivity, even in the absence of an allergen. Epidemiological studies have demonstrated that IgE response is related not only to allergy but also to asthma symptoms, in the presence or absence of atopy, as well as exposure to cigarette smoke. IgE response is altered in several respiratory diseases including extrinsic and intrinsic asthma and allergic bronchopulmonary aspergillosis. CONCLUSION AND PERSPECTIVES: Since anti-IgE monoclonal antibodies are now available for administration to humans, a better understanding of the IgE response may allow the identification of novel therapeutic targets in the field of respiratory disease. PMID- 16249757 TI - [Avulsion of the optic nerve: two case studies]. AB - Avulsion of the optic nerve is a rare and serious injury. The authors report two cases of optic nerve avulsion. The first one concerns a 5-year-old boy who presented ocular trauma after falling on the handlebars of a bicycle. His visual acuity was light perception in the right eye, and his right pupil was unresponsive to light. The anterior segment was normal. The ophthalmoscopic examination showed a total separation of the optic nerve head from the sclera with peripapillary hemorrhage. The second case concerns a 30-year-old man who was hit in the right eye with a stick. On admission, he had no light perception in the right eye, a right afferent pupil defect, a small laceration of the right lower eye lid and no abnormalities on the anterior segment. The fundus examination showed a mild vitreous hemorrhage. Ocular ultrasonography showed vitreous hemorrhage coming directly from the optic nerve head in a mushroom pattern. A CT scan of the orbit revealed a thickened optic nerve. Color Doppler ultrasonography documented slowing of blood flow in the central retinal artery. The two patients received 1 mg/kg/day of prednisone for 2 weeks. No improvement was noted. Optic nerve avulsion is often caused by sudden and forceful rotation of the eye with tearing of the optic nerve as it exits the globe. The nerve can be partially or totally avulsed. The prognosis is usually poor. PMID- 16249756 TI - [Opacification of a hydrophilic acrylic intraocular lens]. AB - PURPOSE: To report pathological and histochemical analysis of an explanted opacified hydrophilic acrylic intraocular lens. PATIENTS: and method: A hydrophilic acrylic intraocular lens--Orion IFP3D6--was explanted 2 years after routine phacoemulsification because of its opacification associated with significant visual impairment (20/40). The intraocular lens was examined by gross and light microscopy. Full-thickness cut sections of the lens were stained with 1% alizarin red (a special stain for calcium). RESULTS: Microscopic analyses revealed the presence of granules of various sizes distributed in a line parallel to the anterior and posterior curvatures of the lens. The granules stained positively for alizarin red. CONCLUSION: Such opacification has been described with the SC60B-OUV, MDR Inc., which is also a one-piece foldable-design intraocular lens (28% hydrophilic material). The opacification of the intraocular lens occurred over various periods after surgery. Forty percent of these lenses have been explanted. Microscopic and histochemical evaluation of these intraocular lenses revealed the same type of granulation deposits within the lenses. The origin and mechanism of the opacification are still not fully understood. PMID- 16249758 TI - [Craniofacial fibrous dysplasia: a case report]. AB - Fibrous dysplasia is classified among benign bone tumors. It is a bone development anomaly characterized by hamartoma proliferation of fibrous tissue within the medullary bone, with secondary bony metaplasia, producing immature, newly formed and weakly calcified bone, without maturation of the osteoblast. It can be monostotic or polyostotic. The craniofacial localization occurs in 10%-25% of cases in monostotic forms and in 50% of cases in polyostotic forms. Fibrous dysplasia essentially affects children and young adults, with no sex preference. Its diagnosis is not always straightforward because the functional symptomology is often absent or not specific. Medical imagery is necessary and in difficult cases a bone biopsy. The major complication of the illness remains sarcomatous degeneration. Only symptomatic forms are treated. We report an observation of craniofacial fibrous dysplasia affecting a child and discovery of proptosis, with no other ocular or general functional signs. PMID- 16249759 TI - [Contribution of in vivo confocal microscopy to limbal tumor evaluation]. AB - PURPOSE: To explore tumors of the limbus with a new in vivo confocal microscope and to compare the images to histology results. METHODS: We evaluated three tumors in three patients with the Heidelberg Retina Tomograph II, Rostock Cornea Module. A diagnostic and therapeutic excision with adjunctive cryotherapy was performed for each individual. Confocal microscopy was compared to histopathologic sections. RESULTS: Histology identified two dysplasias and one carcinoma in situ. The main pathological features were visible on our images: cytonuclear atypias, epithelial folds into an inflammatory and vascularized conjunctival stroma, fine vessels perpendicular to the surface, a clear limit with normal epithelium, papillomatous organization, and abnormal keratinization. CONCLUSION: Our preliminary study showed that this type of limbal tumor could be explored using in vivo confocal microscopy. We were not able to determine whether there was a microinvasion. This new method could be a diagnostic aid, especially for atypical lesions and for follow-up because of frequent recurrences. Other studies are necessary to confirm our hypothesis. PMID- 16249760 TI - [Conjunctival tumors in children. A histopathologic study of 42 cases]. AB - PURPOSE: The aim of this study was to describe the clinicopathological characteristics of 42 conjunctival tumors surgically removed in children. PATIENTS AND METHODS: Records of all conjunctival tumors surgically removed in children during the 11-year period 1990-2001 were collected from the records of the Department of Ophthalmology, Edouard Herriot Hospital, Lyon. In all cases, the tumor was resected with no additional treatment. Sections of all cases were reviewed by the same pathologist. RESULTS: A total of 42 cases (40 patients) were included in the study. The mean age of the subjects at the time of surgical excision was 10 years, with a range of 1-17 years; 45% were male. The most frequent indication for tumor removal was suspected growth. The clinical diagnosis was accurate in 91% of cases. The tumor was localized at the limbus in 57% of cases. Most tumors were pigmented nevi, accounting for 83% of the lesions. The histopathological diagnoses in decreasing order of frequency were: nevi (35), angioma (2), dermolipoma (2), dermoid (1), papilloma (1), and squamous cell carcinoma (1). In one case, the histological study revealed squamous cell carcinoma in a child suffering from xeroderma pigmentosum. CONCLUSIONS: This retrospective study suggests that malignancy of pigmented conjunctival lesions is extremely rare in children. Although squamous cell carcinoma is rare in children, surgical excision and histological study are necessary when a conjunctival lesion is associated with xeroderma pigmentosum or immunodeficiency. PMID- 16249762 TI - [Results of treating uveal melanoma with proton beam radiation: 10-year follow up]. AB - PURPOSE: We analyzed the long-term results of uveal melanoma treatment with proton beam irradiation in a series of patients with a follow-up of at least 10 years. PATIENTS AND METHODS: The patients were treated with proton beam radiation between September 1991 and December 1992. They had an initial examination including visual acuity, funduscopy, A and B scan ultrasonography of the eye, fundus photographs and fluorescein angiography. General examination included chest radiography and B scan ultrasonography of the liver. All tumors received a total dose of 60 cobalt-Gray equivalents (applied in four daily fractions) at the Orsay proton therapy center. RESULTS: A total of 167 patients were treated with a median follow-up of 116 months. Their median age was 59 years. Thirteen tumors were anterior to the equator, 76 overlapped the equator and 78 were posterior to the equator. An initial retinal detachment was present in 41 cases. The optic disk was invaded in 10 cases. The median tumor diameter was 12 mm and the median tumor thickness was 5.8 mm. The mean initial acuity was 20/50. The survival rate was 62.93% at 10 years; 72.9% of deaths resulted from metastasis. Statistically significant risk factors for death identified in the multivariate analysis were tumor diameter greater than 12 mm (p=0.0004) and age over 60 years (p=0.0001). The metastasis rate at 10 years was 31%. The liver was affected in 97.8% of these patients. Risk factors for metastasis were the anterior site of the tumor, its volume greater than 0.4 cc and the presence of retinal detachment at diagnosis. The secondary enucleation rate at 10 years was 13.23%, mainly attributable to secondary neovascular glaucoma. The local recurrence rate was 6%. The visual acuity rate in 42.1% of patients was better than 20/100 at 10 years. Visual loss was mainly due to postradiation maculopathy and neuropathy. CONCLUSION: Our study confirms the long-term results found in the literature on proton beam radiation. This therapy allows good tumor control, an excellent eye retention rate, and good final visual acuity for approximately half of the patients. PMID- 16249761 TI - [Long-term effects of iterative diving on visual field, color vision and contrast sensitivity in professional divers]. AB - PURPOSE: To assess long-term functional effects of iterative diving, we studied visual field, color vision, and contrast sensitivity in 21 French Navy professional divers and 21 controls. PATIENTS AND METHODS: This retrospective study investigated a population of 21 divers and 21 controls. All subjects were male. The inclusion criterion for divers was a total number of dives greater or equal to 1,000. Exclusion criteria for the two groups were glaucoma, ocular hypertension, smoking, and vasospastic risks. Additional exclusion criteria for controls were any history of diving practice and of hyperbaric oxygen therapy. The visual field was examined with a Humphrey Central 30-2 threshold test. Moreover, we explored spatial contrast sensitivity using Metrovision Moniteur Ophtalmologique "STAT" program and color vision with desaturated 15 hue test. RESULTS: None of the divers had any loss of spatial contrast sensitivity. There was a high frequency of yellow-blue axis color vision defects (45.2%) in the diver group. Regarding visual field, corrected pattern standard deviation was significantly higher in divers (p<0.01). CONCLUSION: These findings suggest that iterative diving may cause subclinical functional effects on vision. Further studies will be needed to determine the exact setting of this repercussion upon macula and/or the optic nerve. PMID- 16249763 TI - [Epidemiology of ocular and orbital lesions referred to an ocular oncology center]. AB - PURPOSE: To study the epidemiology of ocular and orbital lesions referred to an ocular oncology center. MATERIAL AND METHODS: Prospective study of ocular lesions referred between November 1997 and November 2002. RESULTS: A total of 1,257 lesions were recorded in 640 females and 617 males. The mean age at the initial visit was 52 years (range, 0 to 94 years). Patients more than 60 years old accounted for 46% of our series. The lesions were classified into four groups: intraocular lesions (69%), superficial layer lesions (18%), adnexal tissues lesions (7%) and orbital lesions (6%). The posterior uvea was the most frequent localization. The clinical diagnosis of 374 lesions (30%) was confirmed by histological examination. The most frequent tumors were retinoblastoma in children and teenagers, conjunctival nevus young adults, choroidal melanoma in slightly older patients and posterior choroidal nevus in elderly people. CONCLUSION: This study reviewed the epidemiological characteristics of ocular and orbital lesions referred to an ocular oncology center for their diagnosis, treatment and follow-up. PMID- 16249764 TI - [Aniridia: five case studies]. AB - INTRODUCTION: Aniridia, or congenital absence of the iris, is a rare and severe ocular abnormality. We report five cases diagnosed in our department over the last 5 years and describe our clinical findings and attitude. OBSERVATIONS: The patients were two adults and three children, 8-45 years old at the time of diagnosis. Visual acuity ranged from light perception to 20/50. Aniridia was associated with other abnormalities in four cases: lens ectopy in one case, ptosis in three cases, microcornea in one case, macular hypoplasia in three cases and a pterygium coli associated with a pectus excavatum in one case. All patients had complications: corneal dystrophy in three cases, lens opacities in five cases, elevated intraocular pressure (IOP) in two cases, severe amblyopia with nystagmus or strabismus in three cases. Only patients with glaucoma received IOP lowering medications. IOP was normalized in both cases and regular controls were instituted for all the other patients. DISCUSSION: Management of aniridia is complex in our regions because patients often consult late, at a stage that involves complications and amblyopia. First-intention surgical treatment of glaucoma is not always justified. CONCLUSION: Only genetic counseling, a close follow-up after an early diagnosis and adapted surgical techniques can improve the visual prognosis of aniridia patients. PMID- 16249765 TI - [Cystic fibrosis and uveitis in childhood and multiple sclerosis at adulthood]. AB - We report the case of a patient with cystic fibrosis who suffered severe bilateral uveitis, with hypopyon, retinal vasculitis, and polyarthritis when he was 6 years old. No etiology could be found. Multiple sclerosis began when he was 22 years old. This panuveitis was clinically very different from the uveitis usually associated with multiple sclerosis. Polyarthritis and skin vasculitis have been reported in the course of cystic fibrosis, but no uveitis nor retinal vasculitis have been described. This raises the question of the role of multiple sclerosis and cystic fibrosis in the pathogenesis of this case of uveitis. PMID- 16249766 TI - [Uveitis and chronic intestinal inflammatory diseases: three case studies]. AB - INTRODUCTION: The most frequent inflammatory chronic bowel diseases are Crohn disease and ulcerative colitis, which present extra intestinal manifestations, particularly, arthritis and ocular inflammation. Anterior uveitis is the main ocular manifestation and remains exclusively an inflammatory process. MATERIAL AND METHODS: [corrected] We report three cases of bilateral uveitis, with ulcerative colitis in two cases and Crohn disease in the last case. In the two first cases, uveitis occurred after 8 years of bowel disease which was stabilized by sulfasalazopyridine and 6 months after bowel surgery. In the third case, the patient was followed for Crohn disease for 3 years with anterior uveitis relapse concomitant to bowel relapse. RESULTS: In the two first cases, there was no recurrence after corticosteroids topical therapy. In the third case, topical and systemic corticosteroids provided adequate control of ocular inflammation, but the follow-up was marked by pupillary and inflammatory glaucoma, which required gonio-surgery. DISCUSSION: Anterior uveitis is the most frequent ocular manifestation in chronic inflammatory bowel disease: approximately 1.9-4.9% in Crohn disease and 1.6% in ulcerative colitis. The uveitis course is synchronous with bowel relapse and its treatment requires anti-inflammatory agents. Nevertheless, in severe relapse, recovery is obtained after the resection of the entire involved bowel segment. PMID- 16249767 TI - [Langerhans-cell histiocytosis of the orbit. A case study]. AB - PURPOSE: Langerhans' cell histiocytosis is a rare disease representing less than 1% of orbital tumors. METHODS: We report a case of Langerhans cell histiocytosis with orbital involvement in a 9-year-old boy. He presented with an inflammatory swelling if the left lateral orbital wall. The computed tomographic scan revealed an orbital cellular mass with lytic bone lesion within the orbital roof and intracranial enlargement. RESULTS: The cytological study after a biopsy showed infiltrates of histiocytes derived from Langerhans cells. Diagnosis was confirmed by immunohistochemistry, which identified positive staining with anti-S100 and anti-CD1a antibodies. The rapidly expanding orbital tumor, posing a threat of ocular compression as well as intracranial spreading, was treated by chemotherapy (Vinblastine) combined with a steroid. CONCLUSION: A 2-year follow-up showed no evidence of recurrence or systemic involvement. According to this observation, the authors describe the clinicopathological and histological features of orbital involvement in Langerhans cell histiocytosis. PMID- 16249768 TI - [Malassezia fungal blepharitis: a case report]. AB - PURPOSE: Blepharitis is a very common disorder but the role of certain mycoses in the etiopathogenesis of blepharitis remains controversial. OBSERVATION: We report the case of a 29-year-old man presenting with bilateral redness and intense itching of the edge of the eyelid. Slit-lamp examination showed vasodilation of the vessels of the edge of the eyelid associated with seborrhea and a sticky yellowish substance at the roots of the eyelashes. When the substance dried and fell, microulcerations were seen at the base of the eyelids, characteristic of follicular-seborrheic blepharitis. Otherwise the ophthalmological examination was normal. Treatment with eye drops (antibiotics, antiseptics, corticoids, antihistamines) was tried several times. The blepharoconjunctivitis regressed during these treatments without totally disappearing, recurring after the treatment was discontinued. Bacteriological and fungal exams were conducted, showing numerous Malassezia furfur spores and hyphae. Given that there was no antifungal eye drop preparation on the market, topical ketoconazole (Nizoral) was prescribed. Two weeks later, the condition had cleared, with no recurrence after 5 months of follow-up. DISCUSSION: Malassezia is a saprophyte species that may play a role in certain cases of chronic blepharitis, either through a reaction of intolerance and hypersensitivity or occasional proliferation. Treatment consists in eradicating the causal fungus using an antifungal agent associated with symptomatic treatment. We stress that fungal exams should be conducted whenever blepharitis recurs. PMID- 16249769 TI - [Neuroretinitis, Parry-Romberg syndrome, and scleroderma]. AB - INTRODUCTION: Progressive hemifacial atrophy (Parry-Romberg syndrome) is a rare entity characterized by severe hemifacial atrophy affecting subcutaneous tissue. Its clinical manifestations are mainly neurological and ocular. The most common ocular finding is enophthalmos with eyelid modifications (ptosis, retraction, atrophy). Neuroretinitis remains a rare symptom in this affection. We report here a new case. CASE REPORT: A 22-year-old woman with progressive hemifacial atrophy (Romberg syndrome) is reported. The atrophy had begun 10 years before. At time of presentation, she was suffering from blurred vision in her left eye. She presented with mild enophthalmos associated with eyelid atrophy and loss of cilia. Biomicroscopic examination showed bilateral vitreitis and a typical spectrum of neuroretinitis in her left eye. There was no heterochromia. General examination revealed atrophy of the right part of her body. Etiological investigations showed the presence of antinuclear antibodies and a positive Rose Waaler latex test without an inflammatory syndrome. Corticotherapy was proposed (1 mg/kg/day) with good progression. DISCUSSION: The authors report a new case of progressive hemifacial atrophy (Parry-Romberg syndrome). They emphasize the rarity of this disease, its etiology, which remains controversial, the diversity of its ophthalmologic manifestations, and its relations with scleroderma and autonomic nervous system disorders are discussed. Acute neuroretinitis remains rare and its pathogeny is unknown. PMID- 16249772 TI - [Cutaneous abnormalities of the eyelid and systemic diseases]. AB - Systemic diseases are unknown diffuse inflammatory disorders. They include systemic and metabolic diseases and connective tissue diseases. Among them, lupus erythematosus, scleroderma, polymyositis and dermatomyositis, Sjogren syndrome, Wegener's granulomatosis, sarcoidosis, Vogt-Koyanagi-Harada's syndrome, and amyloidosis are the diseases encountered most frequently that can manifest cutaneous abnormalities of the eyelids. The main eyelid disorders involved in these diseases are described in this paper. PMID- 16249771 TI - [Dermatosis and infectious dermatitis of the eyelid]. AB - Lid infections are very frequent and for the most part are isolated and benign clinical conditions. Rarely they occur during a severe systemic disease and present a poor prognosis. In this review, we describe the most frequent presentations and detail the available medical and surgical treatments. PMID- 16249773 TI - [Benign pigmented lesions of the eyelids]. AB - We describe benign pigmented tumors of the lids, including benign lesions of the epithelium such as papillomas, seborrheic keratosis, and inverted follicular keratosis; precancerous lesions of the epithelium such as xeroderma pigmentosum and actinic keratosis; melanocytic lesions such as Ota nevus, lentigo, and lentigo maligna; and nevi and other lesions simulating a melanoma such as pigmented hidrocystoma, angiomas, and epithelial cyst. For each of these lesions, we describe the clinical and histopathological aspect as well as the therapeutic guidelines. PMID- 16249774 TI - [Adenocarcinoma in a pleomorphic adenoma of the lacrimal gland: a case study]. AB - PURPOSE: To report a patient with an adenocarcinoma in a pleomorphic adenoma of the lacrimal gland. Adenocarcinoma constitutes a distinct group of epithelial malignancies of the lacrimal gland. METHODS: The clinical presentation, workup, surgical approach, and pathological findings were reviewed. RESULTS: A 56-year old man presented with a 3-month history of a lacrimal fossa mass. This patient presented a painless mass in the upper outer eyelid with significant displacement of the globe. The tumor was localized to the lacrimal gland. Radiological investigations showed a round, well-defined lesion in the fossa of the lacrimal gland. We performed a lateral orbitotomy; en bloc resection was accomplished. The final specimen showed an adenocarcinoma in a pleomorphic adenoma. No recurrences were detected during follow-up. CONCLUSION: Although adenocarcinoma has rarely been reported in association with a pleomorphic adenoma of the lacrimal gland, this combination can exist. If the malignant tumor is limited in the pleomorphic adenoma, the prognosis appears to be better than in cases of local extension. Adjuvant therapy, radiation or others, are not useful. PMID- 16249775 TI - [Cogan's microcystic dystrophy]. PMID- 16249776 TI - Europa Medicophysica is indexed by Medline: opening new perspectives for European, Mediterranean and Italian physical and rehabilitation medicine. PMID- 16249777 TI - Domains of research, development and strategic planning in rehabilitation medicine. AB - Rehabilitation systems are almost universally struggling to maintain or improve their standings. In this sense it is very useful to draw strategic plans for research and development taking into consideration the characteristics of the profession and, at the same time, the environment where they perform. Five domains of activities can be identified in this sense: measurement of clinical outcomes (including cost/effectiveness and quality assurance), services delivery: the continuum of care, technologies/procedures, pharmacological treatments and opportunities (''niches''). Each domain is described and the different possibilities analyzed. Each system--regional, national--or even a facility should make a selection of what fits their strategic plan best and which modality can be easily incorporated and where human and material resources exist or can be acquired. The need for the justification of the rehabilitation services looks by now as a must, universally. PMID- 16249778 TI - Certainties and prospects in physical medicine and rehabilitation. PMID- 16249779 TI - Short history of Europa Medicophysica: 41 years of contributions to the scientific roots of the Specialty of Physical and Rehabilitation Medicine. PMID- 16249780 TI - A randomized controlled trial on the efficacy of physical exercise in patients braced for instability of the lumbar spine. AB - AIM: Spinal instability is often disregarded as a cause of chronic low back pain and until now there has been no agreement as to its definition and on its nosologic importance or as to a conservative therapeutic protocol. The authors aim to verify whether possible symptomatological characteristics are reflected in radiological findings and, although there is no univocal opinion on the utilization of orthoses of containment for unstable segments, they also aim to verify their efficacy on pain control and neuromotor performance when employed in isolation or in association with the most reliable rehabilitation techniques. METHODS: Forty-eight patients between the ages of 30 and 50 were entered in the study, selected with special exclusion criteria and appropriately randomized to a group following kinesitherapy (KT) and orthoses (O) (O+KT group) and to a control group (orthoses [O] group); the symptomatological and instrumental characteristics were studied at time intervals 0 (t0), 3 months (t3), 6 months (t6), and 12 months (12t). RESULTS: The samples examined present homogeneous characteristics. Lumbar instability pain is related to the presence of shift and not to hypermobility, when the latter is guided by efficient neuromotor feedback. In the O+KT group, treatment achieves the two-fold results of reducing shift and increasing mobility in the absence of pain. Both groups tend to increase the utilization of a brace over time. Furthermore, in the O+KT group, a marked reduction in the use of medicine is noticed. CONCLUSIONS: Following treatment both groups report a decrease in pain even though the results of neuromotor performance prove to be better in the group following KT. PMID- 16249782 TI - Are some cases of spina bifida combined with cerebral palsy? A study of 28 cases. AB - AIM: This study is a description of a group of children with spina bifida who present with neurological impairments resembling cerebral palsy. Spina bifida is a complex congenital spinal anomaly causing paraparesis. Some children with spina bifida have neurological impairments, which fit into the definition of cerebral palsy. Extensive spasticity discordant with the level of the spina bifida lesion, upper extremity dysfunction and cognitive impairment are suggestive of concomitant cerebral palsy in these cases. The probable etiology for this problem may be neglected hydrocephalus, meningitis or other brain lesions common in spina bifida. METHODS: In this study we have reviewed our cohort of 365 patients and found 28 cases with the above-mentioned findings. Main evaluation parameters used were the Ashworth scale for spasticity, Green and Banks modified classification for hand function, cognitive function, mental status, ambulation and lesion level. RESULTS: Twenty-eight out of 365 children with spina bifida had neurological impairments resembling cerebral palsy. Their mean age was 59.9+/ 41.3 (range, 16.8-31.2) months. Seventy percent of the patients were nonambulatory and therapeutic ambulation only was present in 30% of patients. Seventy percent of the children had spasticity of Ashworth grade 2 or higher. Upper extremity dysfunction, and cognitive impairment were also observed in 80% of the patients involved in this study. The spinal lesion was 60% thoracal and 20% upper lumbar, and none of the cases had signs of spinal tethering. CONCLUSIONS: We believe that this group of children with spina bifida may be regarded as having concomitant cerebral palsy. This fact implies that the management of this group of patients having mixed findings must be changed accordingly. PMID- 16249781 TI - Nosocomial infections in the rehabilitation department. AB - AIM: The patients of a Rehabilitation Department are at high risk of nosocomial infections because they generally have a long term hospitalisation and more and more frequently immune-compromised subjects, like old patients or with chronic illness, are admitted to rehabilitation programs. However, to evaluate the real infectious risk of a Rehabilitation Unit, it is important to consider also that a high number of patients are transferred from other hospitals after a specific therapy of the acute phase of their medical or surgical pathology and so many nosocomial microrganisms previously acquired may spread to a Rehabilitation Unit. METHODS: From January to December 2003 we have performed a screening of the bacteruria among the patients at admittance to the Rehabilitation Unit of S. Orsola Fatebenefratelli Hospital of Brescia (Italy). RESULTS: A significant bacteruria (>100000 cfu/mL) in 28.9% of 114 patients coming from home and in 41.9% of 179 patients transferred from other hospitals without antibacterial treatment has been documented. CONCLUSIONS: These findings confirm the presence of an high number of patients colonized or infected by nosocomial bacteria previously acquired in hospital and underline the need, in addition to specific skill, of wide infectious knowledge among the medical staff of a Rehabilitation Unit. A specific approach to the infectious problem in the Rehabilitation Department in order to reduce the risk of nosocomial infections may be suggested. PMID- 16249783 TI - Guidelines for rehabilitation of children with cerebral palsy. PMID- 16249785 TI - Cruciate paralysis and hemiplegia cruciata: report of three cases. AB - STUDY DESIGN: Report of three cases of cruciate paralysis and hemiplegia cruciata. OBJECTIVE: To stress the importance of upper cervical spine lesions causing neurological symptoms and signs. SETTING: Neuro-orthopedic service, Fukui University Hospital, Japan. RESULTS: Three patients (all females; one with congenital anomaly at the occiput-atlas level, one with assimilation of the atlas, and one with rheumatoid arthritis-related proliferative synovium) had clinical features of cruciate paralysis and hemiplegia cruciata. All three cases underwent decompressive surgeries. CONCLUSION: Neurological symptoms and signs of cruciate paralysis and hemiplegia cruciata should be carefully assessed, and surgical therapy should be based on the pathological condition. PMID- 16249784 TI - Gait training regimen for incomplete spinal cord injury using functional electrical stimulation. AB - STUDY DESIGN: Case series, and repeated assessments of the same individuals. OBJECTIVE: To demonstrate the feasibility and efficacy of a multiweek intervention on walking function in people with chronic, incomplete spinal cord injuries. SETTING: Rehabilitation hospital for spinal cord injury (SCI) in Toronto, Canada. METHODS: A convenience sample of five subjects with chronic, incomplete SCI trained for 12-18 weeks using a new multichannel neuroprosthesis for walking. The following outcome measures were recorded throughout the training period: walking speed, step frequency and average stride length based on a 2-min walk test. Also identified were which walking aids and orthoses subjects preferred to use, and whether they employed a step-to or step-through gait strategy. Follow-up measurements of three subjects were made up to 10 weeks after treatment. RESULTS: All subjects demonstrated significant improvements in walking function over the training period. Four of the subjects achieved significantly increased walking speeds, which were due to increases in both stride length and step frequency. The fifth subject experienced a significant reduction in preferred assistive devices. Follow-up measurements revealed that two subjects walked slightly slower several weeks after treatment, but they still walked significantly faster than at the start of treatment. CONCLUSION: The gait training regimen was effective for improving voluntary walking function in a population for whom significant functional changes are not expected. This application of functional electrical therapy is viable for rehabilitation of gait in incomplete SCI. PMID- 16249786 TI - Intra-radicular lumbar disc herniation: report of two cases. AB - STUDY DESIGN: A case report. OBJECTIVES: To report two cases of intra-radicular disc herniation. SETTING: Kocatepe University Faculty of Medicine, Department of Neurosurgery, Afyon, Turkey, Vakif Gureba Training Hospital, Department of Neurosurgery, Istanbul, Turkey and Yeditepe University Faculty of Medicine, Department of Neurosurgery, Istanbul, Turkey. METHODS: Two cases with intra radicular disc herniation at L2-3 level in one, and at L5-S1 level in the other were presented. Both patients were admitted with complaints of severe leg pain and motor weakness. In both cases, intra-radicular disc herniation was diagnosed during surgery. RESULTS: The patients had uneventful recovery. They had no complaints or neurological deficits 6 and 3 months after surgery, respectively. CONCLUSIONS: The possibility of an intradural disc herniation should be kept in mind for the success of the management of lumbar disc herniation. In some patients who did not benefit from surgery, intra-radicular or intradural disc herniations may be the cause of failure. PMID- 16249787 TI - Diffusion MRI in spinal dural arterio-venous fistula: a case report. AB - STUDY DESIGN: Case report. OBJECTIVES: Brain diffusion magnetic resonance imaging (MRI) and apparent diffusion coefficient (ADC) maps are useful to differentiate vasogenic and cytotoxic oedema during cerebovascular diseases. We investigate spinal cord diffusion MRI and ADC maps before and after treatment in one case of spinal dural arteriovenous fistula. SETTING: University of Bordeaux, France. METHODS: We used spinal cord diffusion MRI. RESULTS: We found a vasogenic oedema that disappears after treatment. CONCLUSION: This case report confirms the interest of spinal cord diffusion and ADC maps to differentiate vasogenic and cytotoxic oedema at the spinal cord level. PMID- 16249789 TI - Long-term efficacy of AMS 800 artificial urinary sphincter in male patients with urodynamic stress incontinence due to spinal cord lesion. AB - STUDY DESIGN: Retrospective analysis. OBJECTIVES: To evaluate long-term efficacy and complications of AMS 800 (American Medical Systems) artificial urinary sphincter (AUS) in treatment of urodynamic stress incontinence (USI) in male patients with spinal cord lesion (SCL). SETTING: London Spinal Injuries Unit, Stanmore, UK and Institute of Urology and Nephrology, London, UK. PATIENTS AND METHODS: A retrospective analysis identified nine males with SCL (five thoracic, three lumbar, one cervical) with USI, who underwent AMS 800 AUS implantation by a single surgeon at a specialist spinal injuries unit. The mean age was 38.2 years (range 27-47 years), with the mean time since injury of 13.8 years (range 6-26 years). In all, seven were complete and two were incomplete SCL (traumatic spinal injury eight, tranverse myelitis one). All implants were inserted with the urethral cuff around the bulbar urethra with a 61-70 cm water pressure reservoir in the retropubic space. Implant activation was carried out at 6 weeks postoperatively. All patients were regularly followed up in outpatient clinics at the interval of 3 months, 6 months and yearly thereafter. An ultrasound examination for the upper tracts and a video-cystometrogram (VCMG) was carried out at 3 months postsurgery and then yearly. RESULTS: The follow-up ranged from 3 to 133 months (mean 70.2 months). All implants were activated successfully with no intra- or immediate-postoperative complications. At activation all patients reported total urinary continence with seven out of nine implants (77%) currently working well. Two patients reported significant recurrent incontinence at 3 month follow-up, one of whom underwent a removal of the entire implant at the end of 3 months and the other was continent after a pump and cuff revision. The implant removed at 3 months was due to erosion and infection while the second was removed at 24 months due to secondary implant infection. Three out of seven (43%) successful implants required one revision each. One patient continues to report minimal leakage only during transfers with no leak demonstrated on postoperative VCMG. One patient with indwelling urethral catheter, two with suprapubic catheter and one voiding on urge have changed their bladder management to intermittent catheterisations postoperatively. All removals and revision procedures were carried out in the first 53 months of follow-up and four out of seven implants (57%) required no revisions. CONCLUSION: On a long term, AMS 800 is a viable option to treat USI in men with SCL. Mechanical revisions are frequent but effective. Strict patient selection, optimum preoperative bladder management and regular follow-up ensure low complication and high efficacy rates in the long term. PMID- 16249788 TI - Determination of changes in blood pressure during administration of sildenafil (Viagra) in patients with spinal cord injury and erectile dysfunction. AB - STUDY DESIGN: Prospective, open-label, comparative study, to assess the effects of sildenafil on blood pressure in a population of patients with spinal cord injury (SCI). OBJECTIVES: To determine the effect of sildenafil on blood pressure in patients with erectile dysfunction secondary to SCI by comparing changes in blood pressure in SCI patients with a neurologic level below T5 versus higher levels. To establish a relationship between the potential hypotensive effect and protective muscle spasm against blood pressure reduction. To assess the effects of age, complexity and duration of SCI on changes in blood pressure. To record any adverse effects occurring during the study. SETTING: Spinal Cord Injury Unit, Insular University Hospital of Gran Canaria, Canary Islands, Spain. SUBJECTS: In total, 22 male SCI patients aged 18 years or older with a history of SCI greater than 3 months in duration. METHODS: Patients with erectile dysfunction secondary to SCI were included in the study, without excluding patients with a neurologic level above 75 or asymptomatic low blood pressure. Patients with specific contraindications for use of the drug were excluded. A personal history was obtained, and the level of injury (ASIA/IMSOP scales of international standards), impairment grade (ASIA impairment scale), spasticity grade (modified Ashworth scale) and baseline sitting and supine blood pressure values were determined. A single dose of 50 mg of sildenafil was administered, and patients remained sitting at 45 degrees . Blood pressure was monitored every 10 min for 4 h and whenever the patient reported symptoms. Any relevant signs and symptoms manifested during the study period were also recorded. Analysis of the changes in blood pressure values was performed using a paired t-test in each group of patients according to neurologic level and spasticity grade. RESULTS: A decrease in blood pressure was observed in all patients, although patients with a level of injury at T5 or above and those with a complete SCI showed a less intense decrease (P<0.05). The spasticity grade of the patients was protective against the fall in blood pressure, as it was less significant in patients with grade 3 (P>0.1) than in those with grade 0. Adverse effects were few and transient. None were related to hypotension. CONCLUSION: Sildenafil caused a decrease in blood pressure in SCI patients with a neurologic level of injury above T5 and complete injuries (grade A), but did not have clinical implications in the patients studied. A higher spasticity grade tends to protect the patient from the fall in blood pressure. Age and duration of injury do not appear to influence this decrease. PMID- 16249790 TI - Sir Richard Doll, epidemiologist - a personal reminiscence with a selected bibliography. PMID- 16249791 TI - A phase I/II study of oxaliplatin when added to 5-fluorouracil and leucovorin and pelvic radiation in locally advanced rectal cancer: a Colorectal Clinical Oncology Group (CCOG) study. AB - The purpose of this study was to evaluate the maximum tolerated dose (MTD) and recommended dose of oxaliplatin given synchronously with 5-fluorouracil (5FU), leucovorin (LV) and preoperative pelvic radiation for primary unresectable, locally advanced, rectal cancer. Preoperative pelvic radiotherapy using a three- or four-field technique and megavoltage photons comprised 45 Gy given in 25 fractions, 1.8 Gy per fraction, and delivered with escalating doses of oxaliplatin in combination with low-dose LV and 5FU. Chemotherapy was given synchronously with radiotherapy in weeks 1 and 5. Escalating doses of oxaliplatin (85, 130 and 150 mg m(-2)) were given on days 2 and 30, followed by low-dose LV (20 mg m(-2)) and 5FU (350 mg m(-2)), both given on days 1-5 and 29-33. Surgery was performed 6-10 weeks later. The MTD was determined as the dose causing more than a third of patients to have a dose-limiting toxicity (DLT). Once the MTD was reached, a further 14 patients were treated at the dose level below the MTD. In all, 32 patients received oxaliplatin at the three dose levels, median age 60 years (range 31-79), 24 males and eight females. The MTD was reached at 150 mg m( 2) when four out of six patients experienced DLT. Dose-limiting grade 3 or 4 diarrhoea was reported in two out of six patients at 85 mg m(-2), 5 out of 20 at 130 mg m(-2) and four out of 6 at 150 mg m(-2). Grade 3 neuropathy was reported at 130 mg m(-2) (1 out of 20) and at 150 mg m(-2) (two out of six), and serious haematological toxicity was minimal; one grade 3 anaemia at 150 mg m(-2). In all, 28 out of 32 patients completed all treatments as planned; three had radiotherapy interrupted and three a chemotherapy dose reduction. Four patients did not proceed to surgery due to the presence of metastatic disease (two), unfitness (one) or patient refusal (one). Also, 28 patients underwent surgical resection. Histopathology demonstrated histopathological complete response (pCR) 2 out of 27 (7%), Tmic 3 out of 27 (11%), pCR+Tmic 5 out of 27 (19%), pT0-2 6 out of 27 (22%) and histologically confirmed clear circumferential resection margins in 22 out of 27 (81%). Dose-limiting toxicity with oxaliplatin is 150 mg m(-2) given days 2 and 30 when added to the described 5FU LV and 45 Gy radiation preoperatively. The acceptable toxicity and compliance at 130 mg m(-2) recommend testing this dose in future phase II studies. The tumour downstaging and complete resection rates are encouragingly high for this very locally advanced group. PMID- 16249792 TI - Preclinical pharmacokinetics and metabolism of a novel prototype DNA-PK inhibitor NU7026. AB - In this study we investigated the in vitro time dependence of radiosensitisation, pharmacokinetics and metabolism of NU7026, a novel inhibitor of the DNA repair enzyme DNA-dependent protein kinase (DNA-PK). At a dose of 10 muM, which is nontoxic to cells per se, a minimum NU7026 exposure of 4 h in combination with 3 Gy radiation is required for a significant radiosensitisation effect in CH1 human ovarian cancer cells. Following intravenous administration to mice at 5 mg kg( 1), NU7026 underwent rapid plasma clearance (0.108 l h(-1)) and this was largely attributed to extensive metabolism. Bioavailability following interperitoneal (i.p.) and p.o. administration at 20 mg kg(-1) was 20 and 15%, respectively. Investigation of NU7026 metabolism profiles in plasma and urine indicated that the compound undergoes multiple hydroxylations. A glucuronide conjugate of a bis hydroxylated metabolite represented the major excretion product in urine. Identification of the major oxidation site as C-2 of the morpholine ring was confirmed by the fact that the plasma clearance of NU7107 (an analogue of NU7026 methylated at C-2 and C-6 of the morpholine ring) was four-fold slower than that of NU7026. The pharmacokinetic simulations performed predict that NU7026 will have to be administered four times per day at 100 mg kg(-1) i.p. in order to obtain the drug exposure required for radiosensitisation. PMID- 16249794 TI - The structures of exocyst subunit Exo70p and the Exo84p C-terminal domains reveal a common motif. AB - The exocyst is a large complex that is required for tethering vesicles at the final stages of the exocytic pathway in all eukaryotes. Here we present the structures of the Exo70p subunit of this complex and of the C-terminal domains of Exo84p, at 2.0-A and 2.85-A resolution, respectively. Exo70p forms a 160-A-long rod with a novel fold composed of contiguous alpha-helical bundles. The Exo84p C terminus also forms a long rod (80 A), which unexpectedly has the same fold as the Exo70p N terminus. Our structural results and our experimental observations concerning the interaction between Exo70p and other exocyst subunits or Rho3p GTPase are consistent with an architecture wherein exocyst subunits are composed of mostly helical modules strung together into long rods. PMID- 16249795 TI - Assessment of human exposure to copper: a case study using the NHEXAS database. AB - Copper is an essential trace element and adverse health effects can potentially be associated with both very low and very high intakes. Accurate estimates of inhalation and ingestion (food and drinking water) exposures are therefore needed in order to realistically assess any effects of the distribution of copper intakes within the general population. The work presented here demonstrates an application of a customized subset of the MENTOR/SHEDS-4M computational system (Modeling ENvironment for TOtal Risk studies, employing the Stochastic Human Exposure and Dose Simulation approach, for Multimedia, Multipathway, Multiroute exposures to Multiple co-occurring contaminants. The application utilized data from the National Human Exposure Assessment Survey (NHEXAS) for USEPA Region V as well as from a variety of other available databases. The case study, using a statistical population-based modeling framework, was performed for Eaton County, MI. The results of the simulations, aggregated for six age subgroups of the general population, suggest that food intake is the major pathway for total copper exposure, while drinking water can have significant contributions at the tail of the distribution of intakes. Specifically, it was estimated that over 80% of the county population received potential doses of copper from food that were lower than the Institute of Medicine (IOM) Recommended Dietary Allowance (RDA) value of 900 microg/day. Furthermore, the total combined potential dose from food and water was only about two times greater than the recommended value only for individuals with intakes in the range above the 99th percentile of both food and water intakes. The values were well below the upper tolerable intake value of 10,000 microg/day. The inhalation route consistently acted as only a minor contributor to the total exposure. PMID- 16249797 TI - Video methods in the quantification of children's exposures. AB - In 1994, Stanford University's Exposure Research Group (ERG) conducted its first pilot study to collect micro-level activity time series (MLATS) data for young children. The pilot study involved videotaping four children of farm workers in the Salinas Valley of California and converting their videotaped activities to valuable text files of contact behavior using video-translation techniques. These MLATS are especially useful for describing intermittent dermal (i.e., second-by second account of surfaces and objects contacted) and non-dietary ingestion (second-by-second account of objects or hands placed in the mouth) contact behavior. Second-by-second records of children contact behavior are amenable to quantitative and statistical analysis and allow for more accurate model estimates of human exposure and dose to environmental contaminants. Activity patterns data for modeling inhalation exposure (i.e., accounts of microenvironments visited) can also be extracted from the MLATS data. Since the pilot study, ERG has collected an immense MLATS data set for 92 children using more developed and refined videotaping and video-translation methodologies. This paper describes all aspects required for the collection of MLATS including: subject recruitment techniques, videotaping and video-translation processes, and potential data analysis. This paper also describes the quality assurance steps employed for these new MLATS projects, including: training, data management, and the application of interobserver and intraobserver agreement during video translation. The discussion of these issues and ERG's experiences in dealing with them can assist other groups in the conduct of research that employs these more quantitative techniques. PMID- 16249796 TI - The Washington aerial spray drift study: children's exposure to methamidophos in an agricultural community following fixed-wing aircraft applications. AB - This study characterized exposures of eight children living in an agricultural community near potato fields that were treated by aerial application with the organophosphorus (OP) insecticide, methamidophos (O,S-dimethyl phosphoramidothioate). Exposure monitoring included air and deposition samples in the outdoor community environment, outdoor and indoor air samples at each residence, wipe samples of playground equipment, toys, indoor surfaces, and children's hands, and periodic urine samples. Monitoring occurred prior to, the day of, and 1 day following applications. Methamidophos deposition in the community was very low compared to deposition inside the boundaries of the treated fields. Community air concentrations increased from 0.05 microg/m3 (prespray) to 0.11 and 0.48 microg/m3 (spray day morning and afternoon, respectively), decreasing to 0.10 microg/m3 on the postspray day. Air concentrations outside residences followed a similar pattern; indoor levels did not exceed 0.03 pg/m3. Methamidophos residues were found on playground equipment following applications, but not on indoor residential surfaces. The median hand wipe levels increased from < 0.02 (prespray) to 0.08 microg/sample (spray day), decreasing to 0.05 microg/sample (postspray day). Median concentrations of the primary methamidophos urinary metabolite were 61 microg/l before 1100 hours on the spray day, 170 microg/l after 1100 hours on the spray day, and 114 microg/l on the postspray day. Spray day metabolite levels were correlated with time outside on the spray day (rs = 0.68), with spray day hand wipe levels (rs = 0.67), and with postspray day metabolite levels (rs = 0.64). Postspray day metabolites levels were also positively associated with postspray day hand wipe levels (rs = 0.66). The documentation of children's exposure in this study does not necessarily mean that risks for these children were significantly altered, since nearly all children in the United States are exposed to some level of OP pesticides through dietary intake and other pathways. The association of metabolite levels with time spent outside, and the absence of methamidophos in homes indicates that children's exposures occurred primarily outdoors. PMID- 16249798 TI - PM source apportionment and health effects: 1. Intercomparison of source apportionment results. AB - During the past three decades, receptor models have been used to identify and apportion ambient concentrations to sources. A number of groups are employing these methods to provide input into air quality management planning. A workshop has explored the use of resolved source contributions in health effects models. Multiple groups have analyzed particulate composition data sets from Washington, DC and Phoenix, AZ. Similar source profiles were extracted from these data sets by the investigators using different factor analysis methods. There was good agreement among the major resolved source types. Crustal (soil), sulfate, oil, and salt were the sources that were most unambiguously identified (generally highest correlation across the sites). Traffic and vegetative burning showed considerable variability among the results with variability in the ability of the methods to partition the motor vehicle contributions between gasoline and diesel vehicles. However, if the total motor vehicle contributions are estimated, good correspondence was obtained among the results. The source impacts were especially similar across various analyses for the larger mass contributors (e.g., in Washington, secondary sulfate SE=7% and 11% for traffic; in Phoenix, secondary sulfate SE=17% and 7% for traffic). Especially important for time-series health effects assessment, the source-specific impacts were found to be highly correlated across analysis methods/researchers for the major components (e.g., mean analysis to analysis correlation, r>0.9 for traffic and secondary sulfates in Phoenix and for traffic and secondary nitrates in Washington. The sulfate mean r value is >0.75 in Washington.). Overall, although these intercomparisons suggest areas where further research is needed (e.g., better division of traffic emissions between diesel and gasoline vehicles), they provide support the contention that PM(2.5) mass source apportionment results are consistent across users and methods, and that today's source apportionment methods are robust enough for application to PM(2.5) health effects assessments. PMID- 16249800 TI - The ecology of inbreeding depression. PMID- 16249799 TI - Analyses of magnetic-field peak-exposure summary measures. AB - Two previous epidemiologic studies reported an association between the maximum magnetic field exposure logged during a 24-h measurement period and risk of miscarriage. A hypothesis was put forth which argued that the observed association may be the result of behavioral differences between women with healthy pregnancies (less physically active) and women with miscarriage. We analyzed four existing data sets with power-frequency magnetic-field personal exposure (PE) measurements to investigate the characteristics of peak-exposure measures. We found that the value of the measured maximum magnetic-field exposure varied inversely with the sampling interval between magnetic-field measurements and that maximum values demonstrated less stability over time in repeated measurements, compared to time-weighted average and 95th and 99th -percentile values. We also found that the number of activity categories entered by study subjects could be used to estimate the proportion of subjects with exposure above various threshold values. Exposure metrics based on maximum values exceeding thresholds tend to classify active people into higher exposure categories. These findings are consistent with the hypothesis suggesting that the association between maximum magnetic fields and miscarriage are possibly the result of behavioral differences between women with healthy pregnancies and women who experience miscarriages. Thus, generalization from a given study to more global exposure characterization should be made with particular caution and with due consideration to sampling interval and other characteristics of the measurement protocol potentially influencing the measured maximum. Future epidemiologic studies of peak magnetic field exposure and spontaneous abortion should carefully evaluate the potential confounding effect of the women's activity level during pregnancy. PMID- 16249801 TI - Genome evolution: micro(be)-economics. PMID- 16249802 TI - Gene flow and hybridisation in a mixed oak forest (Quercus pyrenaica Willd. and Quercus petraea (Matts.) Liebl.) in central Spain. AB - Oaks are long-standing models for the study of gene flow and hybridisation. Temperate (Quercus petraea) and sub-Mediterranean (Quercus pyrenaica) oaks coexist in central Spain, showing remarkable differences in population size and structure. Q. petraea has a scattered distribution in central Spain, where it is at one of the southernmost limits of its range, and forms low-density stands; in contrast, Q. pyrenaica is widespread in the region. We selected a mixed population of the two species ( approximately 13 ha, 176 adults and 96 saplings) to compare the patterns of gene flow within each species and the extent of introgression between them. Using five nuclear microsatellite markers, we performed a parentage analysis and found considerable immigration from outside the stand ( approximately 38% for Q. petraea and approximately 34% for Q. pyrenaica), and estimated average seed-dispersal distances of 42 and 14 m for Q. petraea and Q. pyrenaica, respectively. Introgression between species was also estimated using our microsatellite battery. First, we developed a multivariate discriminant approach and, second, we compared our results with a widely used clustering method (STRUCTURE). Both analyses were consistent with a low level of introgression between Q. petraea and Q. pyrenaica. Indeed, only 15 adult trees, approximately 8.5%, were identified as putative hybrids when both methods of analysis were combined. Hybrids may be most common in contact zones due merely to physical proximity. PMID- 16249804 TI - [Trends in cancer mortality and morbidity in Hungarian and international statistics. Characteristics and potential outcome of public health screening programs]. AB - This work is a comparative analysis of Hungarian and international cancer mortality and morbidity data with special attention to the epidemiological background of these indices. The authors also discuss the epidemiological reasons for a national public health screening program, its major objectives and the strategy of choice in relation to similar international programs. The recent cancer mortality and morbidity data including their trends are also provided. PMID- 16249805 TI - [Results of monitoring the year 2002 of the Hungarian breast cancer screening program]. AB - AIM: The aim of the study is to analyse the regional and time range characteristics of the breast cancer screening programme and the utilization of health services related to the programme. METHODS: The data derive from the database of the National Health Insurance Fund Administration containing routinely collected financial data. The patients include all the women having mammography screening in the year of 2002 (N=314,395). In the time range analysis the starting point (T0) was the time of the mammography screening identified with the outpatient code "42400 mammography screening". We calculated the average delay between the time of mammography screening (time=T0), further diagnostic (time=T1) and therapeutic (time=T2) procedures. For the calculation of the average period spent from the time of mammography screening we used the median value instead of arithmetic mean. RESULTS: According to our data 17,303 women had ultrasound examination in axilla (T1 median value: 20 days) and 23,249 women had ultrasound examination in breast (T1 median value: 26 days). Among the women having mammography examination in 2002, 906 had chemotherapy (T2 median value: 83 days), while 1364 patients had radiotherapy (T2 median value: 136 days). The T2 median value of subtotal and total mastectomy was 43-47 days and 50-53 days respectively, while the T2 median value of breast operations because of non malignant causes was 57 days after mammography screening. The total annual cost of organised breast cancer screening programme, including the cost of mammography examination, the cost of further diagnostic examination and surgical, radio- and chemotherapy treatment of recalled women, was 2,242 billion Hungarian forints (8,968 million euros) in 2002. CONCLUSION: We observed significant regional differences, which result in large discrepancies in the equity. We can assume that these differences can be reduced by better organisation and the more consistent application of professional guidelines. PMID- 16249806 TI - [Spatial analysis of mortality caused by malignant tumors, especially by digestive system tumors, using empirical Bayes estimates in Fejer County, Hungary]. AB - General practitioners in the vicinity of Lake Velencei in Fejer County, Hungary, have reported an unusual pattern of recent cancer mortality. Our aim was to clarify whether the presumed mortality cluster is epidemiologically justified; whether it is restricted to the locations in question or also appears elsewhere in the county; and if it is associated with some particular disease group. County mortality from malignancies of the digestive system, other malignancies, and all other causes for the period 1994 to 1999 was analyzed in 15- to 64-year-old men and women, using conventional standardized mortality ratios and empirical Bayes estimates. A continuous surface was interpolated from settlement level data for mortality maps. A mortality cluster from men's digestive cancers is apparent north and east of Lake Velencei and also elsewhere in the county. Differences from the country average in the frequency of males' deaths from other malignancies are fairly limited. A number of problematic areas in men's mortality from other causes are identifiable, including some of the settlements under the primary focus. Women's digestive tract cancer mortality in the area of the men's cluster near Lake Velencei is below the national average. There are almost no differences from the country level in women's deaths from other malignancies. Female mortality from all other causes shows remarkable elevations in the south of the county. Our results suggest the possible role of geographically localized exposures. Men's high mortality from digestive tract cancers is a problem affecting a considerable area of the county, necessitating further investigation. Continued search for causes is also warranted by some estimates of exceptionally high death rates in women from causes other than malignancies. PMID- 16249807 TI - [Primary lymphoma of the heart]. AB - The authors describe the case of a 59-year-old woman, treated for breast cancer by mastectomy and chemotherapy 13 years before her death. Eleven years later she was treated successfully by gastric resection and chemotherapy for gastric cancer. In the last five months, the patient presented dyspnoea, leucopenia, hydropericardium and thoracic fluid both sides. In vivo the origin of these symptoms has not been discovered, neither by cytology nor by pleural biopsy. PMID- 16249808 TI - [Therapy of brain metastasis based on the RPA classification]. AB - RPA classification of patients suffering from brain metastases is not widely used in Hungary. The authors reviewed the RPA disposition-based therapeutic recommendations in the literature. Retrospective analysis of their 123 brain metastatic cases showed 3.8 months median, 34.1% 1-year and 7.9% 2-year overall survival. Patient number and median survival in subgroups: RPA 1: 42/14 months, RPA 2: 38/6,2 months, RPA 3a: 6/3.1 months, 3b: 13/2 months, 3c: 10/0.7 months. Median survival of patients with brain metastases from cancer of unknown primary (CUP) was 3 months. In RPA class 1 and 2, 10% undertreatment has been found for solid brain metastases, and all of the 3c patients were over-treated according to literature recommendations. The authors strongly recommend the use of RPA classification in the management of brain metastases and in contemplation of the capacity of radiotherapy/neurosurgery and oncology. PMID- 16249809 TI - [Complex diagnosis of follicular lymphomas]. AB - INTRODUCTION: In the Western world the second most common type of non-Hodgkin's lymphomas is follicular lymphoma (FL) comprising 30-35% of all cases. According to the data of the National Cancer Registry and our institute, this ratio is lower in Hungary and is about 15-20%, but the occurrence shows an increasing tendency. AIMS: Our aim was to survey and revise FLs that had been diagnosed at the National Institute of Oncology between 1990-1995. We studied the diagnostic relevance of histology, immunohistochemistry and the detection of immunoglobulin heavy chain (IgH) and bcl-2 gene rearrangements. MATERIALS AND METHODS: We surveyed 53 cases that were previously diagnosed as follicular or centrocytic, centroblastic lymphoma. Following histological re-examination, immunohistochemistry (CD20, CD3, bcl-2, CD10, bcl-6, CD5, p53, cyclin D1 and Ki 67) was performed on each case. We also studied the IgH and bcl-2 (major breakpoint region=MBR) gene rearrangement on paraffin embedded samples with conventional PCR methods. The classification was made according to the new WHO classification. RESULTS: After the revision of the 53 cases we found 37 follicular, 11 diffuse large B-cell, 1 mantle cell and 1 marginal zone lymphomas, 1 nodular lymphocyte predominant Hodgkin's lymphoma and 2 follicular hyperplasias. The grade of the FLs correlated with the expression of different antigens. CD10, bcl-2 expression and the proliferation index with Ki-67 showed good correlation with the grade of FLs. We could detect bcl-2 gene rearrangement in 55% of the FLs. CONCLUSION: Considering the diagnostic relevance of the different methods we can conclude that histology alone is not sufficient to make a correct diagnosis. Ninety percent of our cases were solvable with the help of immunohistochemistry and in 10% of the cases the diagnosis was partly based on the molecular pathological results. PMID- 16249810 TI - [Identification of 3q21q26 syndrome by "multipoint" interphase FISH analyses in childhood myeloid leukemia]. AB - Cytogenetic syndrome involving bands 3q21 and 3q26, known as "3q21q26 syndrome" has been observed in adult patients with acute myelogenous leukemia (0.5-2%), chronic myelogenous leukemia in blast crisis (20%), myelodysplastic syndromes and myeloproliferative disorders. In the present study bone marrow samples from two boys (12 and 16 years), diagnosed with CML and AML respectively, were investigated using conventional cytogenetic methods, interphase "multipoint" fluorescence in situ hybridization (FISH), dual color-FISH and multiplex FISH. The "multipoint" FISH analysis identified in de novo childhood AML case an inv(3)(q21q26) and a complex 3q rearrangement including inversion and duplication in the CML case. The "3q21q26 syndrome" is associated with normal or elevated platelet counts with marked abnormalities of megakaryocytopoiesis, involvement of multiple hematopoietic lineages. The affected patients were resistant to conventional chemotherapy and had a short survival. This syndrome is very rare in de novo childhood AML, and simultaneous presence of 3q inversion and duplication, to our knowledge, has not yet been identified in hematological malignancies. The results of our study emphasize the importance of classical and modern cytogenetic analysis in the diagnosis of hematologic malignancies, because in the majority of cases they can provide additional diagnostic information for the clinicians in deciding the best therapeutic approach, precise classification and prognosis of the disease. PMID- 16249812 TI - [Clinical signs and differential diagnosis of iris melanoma]. AB - Iris melanoma is the rarest type of uveal melanomas. Only 4-5% of uveal melanomas occur on the iris. Although the iris can be easily examined due to its location, differentiation of melanocytic malformations such as naevi or melanomas is difficult for the examiner. According to publications by Rones and Zimmermann, histological examinations showed 22% of tumors to be malignant and 78% to be benign. This lead to iridectomy and iridocyclectomy as therapeutic solutions to gain ground over enucleation. Follow-up of the clinical signs, transillumination, ultrasonic biomicroscopy, iris fluorescein angiography and photo-documentation of the clinical signs can be of great help in diagnosis of pigmented iris tumours. Growth of the tumour, secondary glaucoma, hyphaema, significant vascularisation of the tumour and increasing extent of pigmentation can be signs of malignant behaviour. PMID- 16249811 TI - [Effects of 1800 MHz GSM-like exposure on the gonadal function and hematological parameters of male mice]. AB - The aim of this study was to evaluate the possible effects of in vivo 1800 MHz GSM-like exposure on male reproduction. In five separate experiments, male NMRI mice (35-41 g) were exposed (11-12 mice each) to 1800 MHz GSM-like radiation. The average power density was 100 microW/cm2, the estimated SAR was 0.018-0.023 W/kg. The animals were exposed ten times (over two weeks on workdays) and the duration of exposure was 2 h/day. On the day of the last treatment, mice were anesthetized with i.p. pentobarbital, and blood samples were taken for hematology, serum chemistry and serum testosterone (T) determinations (ELISA). Testicles, epididymes, adrenals, prostates and pituitary glands were removed for histology. One testicle of each animal was used for culture of Leydig cells. The cells were cultured for 48 h in the presence or absence of human chorionic gonadotropin (hCG) to evaluate the in vitro steroidogenic response of Leydig cells. In the exposed animals red blood cell count (RBC: 8.59+/-0.10 T/l, n=37) and volume of packed red cells (VPRC: 42.29+/-0.43%, n=37) were significantly higher (p<0.01) compared with the controls (RBC: 8.12+/-0.08 T/l, n=36; VPRC: 39.76+/-0.36%, n=36). The serum testosterone level of the exposed animals (7.85+/-1.08 ng/ml, n=56) was also significantly elevated (p<0.05) compared to the controls (5.12+/ 0.79, n=52), while the in vitro steroidogenic capacity of the Leydig cells was unaltered. No significant differences in the other investigated variables were found between controls and exposed mice. Our results indicate that the applied GSM-like microwave exposure may induce slight, but statistically significant alterations in some hematological and endocrine parameters of male mice within the physiological range. Further investigations are required to establish the biological significance of these phenomena. PMID- 16249813 TI - [The role of neoadjuvant therapy in the treatment of locally advanced, stage III non-small-cell lung cancer]. AB - OBJECTIVE: to review the progress achieved in the neoadjuvant treatment of patients with locally advanced NSCLC by evaluating the articles published during the last 10 years. METHOD: altogether 51 articles (including prospective and retrospective studies, and reviews) were analysed with the intention to evaluate the efficacy of the different neoadjuvant modalities. RESULTS: different types of neoadjuvant treatments were reported. The articles were heterogeneous not only in the aspect of the patient populations but also in regard to the treatment modalities and schedules, the mode and timing of combination of chemotherapy with radiotherapy. Most studies support the advantage of chemoradiotherapy. The chemotherapy should be platinum based, and combination with modern drugs as gemcitabine is recommended. Surgery provides the best local control of the disease. Lobectomy, and in special circumstances sleeve resection, are preferred. The best prognostic factors are the R0 resection and the downstaging of the mediastinal lymph nodes. The cause of death is usually progressive disease, often brain metastases. Because of frequent occurrence of brain metastases, preventive cranial irradiation is recommended by many authors. CONCLUSION: to be able to compare different treatment modalities with consistent patient groups, patients should be classified into well-defined subgroups. T4 tumours (except pleuritis carcinomatosa) behave as locoregional disease, N2/N3 tumours, however, resemble more to systemic diseases. With two- or three-modality treatment, 3-7% improvement can be achieved in the 5-year survival time of patients. Evaluating the results, the heterogeneity of stage III NSCLC should always be kept in mind. It is still unknown to which patients should surgery be offered after neoadjuvant therapy, and who will benefit more from chemoradiotherapy alone. Considering the downstaging effect of the therapy, the tumour and nodal state should be evaluated separately. To answer these questions, it is time to start large randomised studies. PMID- 16249814 TI - [Local relapse in young (< or = 40 years) women with breast cancer after mastectomy or breast conserving surgery: 15-year results]. AB - PURPOSE: Data from various sources indicate that after breast conserving surgery (BCS), younger patients have a high risk of local relapse, but there is insufficient evidence about the risk of post-mastectomy local recurrence. This study investigates the risk of local recurrence for young (< or = 40 years) patients treated with either conservative or radical surgery, with or without radiotherapy (RT). METHODS: 148 young (< or = 40 years) women with early invasive breast cancer underwent axillary dissection and mastectomy (n = 92) or BCS (n = 56) between January 1983 and December 1997. When adjuvant RT was given, the median dose was 50 Gy. The risk factors of local recurrence were estimated by uni and multivariate analysis. RESULTS: At a median follow-up time of 199 months 60 (40.5%) women died of breast cancer. The type of surgery (mastectomy vs. wide tumour excision) had no significant impact on breast cancer-specific survival. The crude rate of local relapse for nonirradiated mastectomy and BCS patients was 24% and 75% (p = 0.0041), and for irradiated patients 4% and 23%, respectively (p = 0.0091). After mastectomy in univariate analysis nodal status (negative vs. positive) and RT (no vs. yes) were significant predictors of local control, but tumour size (T1 vs. T2) and histological grade (1-2 vs. 3) were not. In multivariate analysis both nodal involvement and omission of RT remained independent significant negative predictors of local control. After BCS in univariate analysis extensive intraductal component (EIC, negative vs. positive) and RT (no vs. yes) were significant predictors of local control, but tumour size (T1 vs. T2), nodal status (N0 vs. N1) and histological grade (1-2 vs. 3) were not. In multivariate analysis omission of RT and presence of EIC remained independent significant negative predictors of local control. The 15-year actuarial rate of local relapse was 29% for irradiated, and 75% for nonirradiated BCS patients (RR, 0.21; 95% CI, 0.07-0.55; p = 0.0052). The 15-year actuarial rate of local recurrence was 6% for irradiated, and 46% for nonirradiated node positive mastectomy patients (RR, 0.12; 95% CI, 0.06-0.96; p = 0.0095). CONCLUSION: The incidence of local recurrence is high for young patients treated either with BCS or mastectomy, and RT significantly reduces the risk. The use of postmastectomy RT in node-positive patients gives a good local control. The efficacy of BCS and RT as a treatment modality for young patients needs further investigations. PMID- 16249815 TI - [Radiotherapy of lung cancer: survey and latest evidences]. AB - Based on their experience, the authors survey Hungarian and international literature for changes of indications, techniques and results in lung cancer radiotherapy. At the end of historical review, the latest evidence-based levels of different indications, the up-to-date techniques and the expected results are discussed. PMID- 16249816 TI - [Lens-sparing external beam radiotherapy of periocular lymphomas]. AB - PURPOSE: The presentation of techniques and results of lens-sparing external beam radiotherapy of patients with ophthalmologic lymphomas. MATERIAL AND METHODS: From 1991 to 2001, at the Department of Radiotherapy of the National Institute of Oncology, Budapest, 92 patients with periocular lymphomas were treated by photon and/or electron beam irradiation depending on the localization. Mean age was 63 years (range: 26-89 years) and the male/female ratio was 1.3. According to the Ann Arbor classification, 72% were in stage I.E. Histologically 85 patients (92.4%) were Grade I while 7 patients (7.6%) presented with higher grade disease. In case of low-grade lymphoma, a dose of 24-32 Gy was delivered, and for higher grade of malignancy we applied 34-40 Gy total dose, using 1.8-2 Gy daily fractions. RESULTS: The local remission rate was 94.4% (CR: 78.8% and PR: 15.6%). There was no change in 2.3%, and 3.3% showed progression. Mild and intermediate acute side effects occurred in 28%, and the incidence of chronic late complications was 3%. CONCLUSION: External beam radiotherapy applying lens sparing methods is safe and effective in the treatment of periocular lymphomas. PMID- 16249818 TI - [Improvement of the first cerebral stereotactic radiosurgery system of Hungary]. AB - OBJECTIVE: The first cerebral stereotactic radiosurgery system in Hungary was built in 1991. This system was based on a Leksell stereotactic head frame and a Neptun 10p linear accelerator. We performed 624 radiosurgery treatments with this system between 1991 and 2000. Our objective was to increase the reliability of operation and to extend the applicability of our radiosurgery system. METHODS AND MATERIALS: We modified our stereotactic floor stand with specially designed adapter plates to make it compatible with the Mevatron KD and Neptun 10p linear accelerators and other stereotactic head frames (Riechert-Mundiger, CRW and BrainLab). We made a new tertiary collimator holder attachable to the Mevatron KD linac. The range of treatable cerebral lesion was increased from 10-30 mm to 5 42.5 mm with additional collimator inserts. With the above modifications our radiosurgery system is compatible simultaneously with the Neptun 10p and the Mevatron KD linear accelerators. This way we were able to increase the reliability of operation of the system, as the treatment can be performed with the Neptun 10p linac in case of breakdown of the Mevatron KD linac after fixation of the head frame to a patient's skull. RESULTS: The measured diameter of the radiation isocenter defined by the new radiosurgery collimator was less than 1 mm with the Mevatron KD linac. According to the Lutz-test the distance between the radiosurgery isocenter and the rotation axis of ZIV treatment table was less than 0.5 mm. Results of phantom test showed that the overall spatial precision of our modified radiosurgery system was better than 1.3 mm with Leksell head frame. CONCLUSIONS: On the basis of experiences with 662 patients' radiosurgery treatments, the extension of our first cerebral radiosurgery system to Mevatron KD linear accelerator resulted in a more reliable operation. In accordance with our phantom tests the extension of the original system did not worsen its overall spatial precision. PMID- 16249817 TI - [Radiotherapy of inoperable brain tumors in childhood]. AB - STUDY OBJECTIVE: Description and evaluation of radiotherapy of inoperable tumours of central nervous system. New possibilities in radiotherapy planning. MATERIALS AND METHODS: Between 1987 and 2004, 58 patients (30 boys and 28 girls, mean age 8.5+/-4.3 years) with brain stem tumours were treated with 6 MV or 9 MV photons of the linear accelerator, or with telecobalt. The doses administered ranged from 30 to 71.2 Gy; mean 51.7+/-8.4 Gy. Treatment in each case was performed according to CT- and/or MR-based radiotherapy plan. As of 2000, in case of 16 patients a 3D conformal irradiation plan has been prepared. RESULTS: All patients were followed. The mean follow-up period was 21.8 months (range: 7 to 158 months). The mean duration of symptoms prior to diagnosis was 6.6+/-8.1 weeks (range: 1-52 weeks). Overall survival (OS) and progression-free survival (PFS) rates for all 58 patients were 39.9% and 21.2% at 1 years, 19.1% and 14.7% at 2 years, and 14.8% and 14.7% at 3 years respectively. The following factors were of prognostic influence on the OS in univariate analysis: duration of symptoms (< or = 2 months vs. >2 months, p=0.0081), radiological response (p < 0.0001) and clinical improvement (p = 0.0003). Prognostic factors for PFS were similar to those predicting for OS. Multivariate analysis revealed that the radiological (OS, p = 0.0034, PFS, p = 0.024) and clinical (OS, p = 0.026, PFS, p = 0.044) improvement 6 weeks after completion of radiotherapy were of prognostic significance. CONCLUSION: Accurate field arrangement, precise patient fixation, and CT- and MR based 3D conformal radiotherapy planning may result in better outcome of disease as well as minimal effect on surrounding normal tissues. Radiotherapy should be provided also for histologically not verified tumours. Sixty-eight percent of these patients had a transitory 20.6-month improvement, and 12.1% of them completely recovered. PMID- 16249820 TI - [Computerized radiotherapy planning: retrospective analysis and current methods]. AB - PURPOSE: To present the development of treatment planning methods at the National Institute of Oncology (NIO) from 1969 till 2005. METHODS: The methods and devices of treatment planning is described chronologically. RESULTS: First we did the treatment planning with in-house made devices: body contour drawing instrument, simplified anatomical cross sections, treatment planning table for the cross section projection, archives of isodose curves adjusted to body contours, etc. It was a significant improvement when the graphical addition of isodose curves was followed by computerized dose calculation. In 1978 the work of the Computerized National Treatment Planning Network was started. The Network was organized by IAEA, Ministry of Health Hungary and NIO. The modern treatment planning started at NIO in 1981. From this year, the treatment planning was based on CT, using the CT apparatus of the Medical Postgraduate University. In 1991 a Siemens MEVAPLAN treatment planning system was installed at NIO. The CT data were transferred to the system via floppy disk. The 3D treatment planning program (Pinnacle software of ADAC) started in 2000. The CT, the treatment planning system and newer linear accelerators are connected through the computerized radiotherapy network. Patient positioning, fixing and control devices (mask, EPID etc.) increased the efficacy of the treatment. In-house made devices help this aim too: mirrors at the linear accelerators, special skin marks for CT, block verification unit, multileaf collimator for x-ray treatment simulator. In this year the intensity-modulated radiotherapy (IMRT) will be started at NIO. CONCLUSIONS: The treatment planning at NIO developed to high degree during the investigated time, and it had a considerable effect on the efficacy of radiotherapy. PMID- 16249819 TI - [Dosimetric assessment of external treatment plans for breast cancer: the significance of dose prescription selection]. AB - PURPOSE: To analyze the treatment plans of breast irradiation performed with two tangential beams, to discuss the importance of dose prescription, and to estimate the differences in delivered dose due to various dose prescription methods. MATERIAL AND METHODS: A survey was performed between the Hungarian radiotherapy centers in order to compare the dose prescription methods. Then, treatment plans of 125 breast cancer patients treated in our department were evaluated. The irradiations were performed with cobalt unit, and with 6 and 9 MV photon beams of linear accelerators. The dose distributions were normalized to izocenter, then dose values in five points in central plane; local medial, lateral and central maximums (D(med), D(lat) and D(cent)); volumetric maximum and its location were determined. To characterize the dose to lung and to heart at left-sided tumors the central lung distance (CLD) and maximum heart distance (MHD) were used. Based on the results, estimation was made to assess the differences between delivered dose due to various dose prescriptions applied at the institutions. RESULTS: Four types of dose prescription are currently used in our country, and most frequently the isocenter is selected as a reference point. In the central plane the calculated dose in all but one points differed only a little from the dose to isocenter. The mean D(med), D(lat) and D(cent) were 107%, 107% and 101%, respectively. The volumetric maximum was on average 13% higher than the dose to isocenter. Regarding the beam qualities, this value was 16%, 13% and 11% for cobalt unit, 6 MV and 9 MV photon beams, respectively. The mean CLD and MHD were 1.9 and 0.8 cm, respectively. The difference between delivered doses at the institutions was 6% on average, but in extreme cases it can be as high as 20%. CONCLUSIONS: Three-dimensional treatment planning and plan evaluation are recommended at breast irradiation, especially for large breasts. Since the various dose prescriptions may result in significant differences in the delivered doses, use of a standard dose prescription protocol is recommended. PMID- 16249821 TI - [New clinical data on erythropoietin therapy of cancer patients with anemia. Summary of the major presentations at the Annual Meeting of the American Society of Clinical Oncology (ASCO) in Orlando, 2005]. AB - Anemia of patients with malignancy might have various reasons. No matter if its background is the underlying tumorous disease or chemo- and/or radiotherapy, it can cause fatigue, malaise, it certainly decreases the patients' quality of life and, furthermore, shortens their survival. Chronic hypoxia caused by anemia promotes tumor progression by several mechanisms e.g. by enhancing angioneogenesis by the production of VEGF. At the same time it decreases the efficacy of chemo- and radiotherapy. Therefore, prevention and/or correction of chemo/radiotherapy-induced anemia is a major goal of modern oncotherapy. PMID- 16249822 TI - An efficient method of identifying major depression and panic disorder in primary care. AB - The research literature is replete with evidence of and concerns about the prevalence and undertreatment of mental disorders in primary care. Although screening, on its own, may not directly affect clinical outcomes, it is still the most efficient and effective way to identify psychologically distressed patients for either research purposes or to provide patients with or refer patients to appropriate care. The current study sought to establish the utility of the MHI-5 for the detection of patients suffering from major depression or panic disorder, two of the most common psychiatric conditions seen in primary care settings. This study was conducted in a family medicine clinic and 246 adult outpatients participated. Patients completed the Mental Health Index-5 (MHI-5) as the screening measure and the PRIME-MD Patient Health Questionnaire (PHQ) as the diagnostic instrument. ROC analyses indicated that a cut-off score of 23 on the MHI-5 yielded a sensitivity of 91% and a specificity of 58% for predicting provisional diagnoses of major depression or panic disorder from the PHQ. Using a single item to screen for a PHQ diagnosis of major depression yielded a sensitivity of 88% and a specificity of 62% and a second question had a sensitivity of 100% and specificity of 63% for PHQ diagnosis of panic disorder. These results indicate that it is possible to use a small number of items to efficiently and effectively screen for mental disorders affecting a significant portion of primary care patients. PMID- 16249824 TI - Polymyalgia rheumatica and colon malignacy: case report. AB - Polymyalgia rheumatica (PMR) is a relatively common disorder in the elderly. Whereas the relationship between polymyalgia rheumatica and giant cell arteritis (GCA) is well recognized, there is still controversy about PMR and malignancy. We are presenting a patient with PMR and adenocarcinoma of the sygmoid colon and hypothesize a paraneoplastic relationship. PMID- 16249823 TI - Technical note: the design of a stereotactic frame for direct MRI-anatomical correlation of the brachial plexus. AB - The purpose of this study was to identify optimal magnetic resonance imaging (MRI) conditions to visualize discrete alterations of brachial plexus components, as part of a biomechanical study of minor nerve compression syndromes. A method was developed allowing direct comparison between the MRI image and the subsequently obtained matching anatomic section of the same specimen. We designed a stereotactic frame to obtain the precise orientation of the MRI plane with reference to the specimen and adapted a vertical band saw for multiplanar sectioning of cadaveric specimens. Two cadaveric upper quadrants were examined by MRI (TR 450 ms, TE 13 ms, pixel matrix 512 x 512 and FOV 23-26 cm) and anatomical slices were produced. One specimen was sectioned axially, while the second specimen was sectioned in an oblique plane corresponding to the natural longitudinal axis of the upper part of the brachial plexus. MR images and the corresponding slices exhibited a strong correlation. This correlation was checked by using vitamin A pearls as landmarks. MR images revealed more detail after the correlating anatomical slices were analyzed. The present study shows that the method is suited for direct MRI-anatomic comparison of the brachial plexus and is also proposed for application to other topographical regions. PMID- 16249825 TI - The alpha 2 type IX collagen gene tryptophan polymorphism is not associated with rheumatoid arthritis in the Japanese population. AB - The aim of this study was to investigate whether the alpha 2 type IX collagen (COL9A2) polymorphism that introduces tryptophan residue into the collagen triple helix is a marker of susceptibility to, or severity of, rheumatoid arthritis (RA). The study included 749 Japanese patients with RA. One hundred twenty-four unrelated healthy individuals served as the control subjects. The relationship between the COL9A2 gene polymorphism and clinical manifestations of RA was evaluated. For the number of subjects positive for COL9A2 tryptophan polymorphism, there was no statistically significant difference between RA patients and normal controls. Furthermore, we did not detect any association of COL9A2 tryptophan polymorphism with disease status, least erosive subset, more erosive subset, or mutilating disease. The lack of association of COL9A2 tryptophan polymorphism with RA and the clinical findings in our study implies that the polymorphism may not function as a candidate gene marker for screening RA patients. PMID- 16249826 TI - Circulating auto-antibodies against nuclear and non-nuclear antigens in primary Sjogren's syndrome: prevalence and clinical significance in 335 patients. AB - The aim of this study was to analyze the prevalence and clinical significance of circulating auto-antibodies against nuclear and non-nuclear antigens in a large cohort of Spanish patients with primary Sjogren's syndrome (SS). We studied 335 patients diagnosed with primary SS seen consecutively in our department since 1994 and tested for anti-nuclear antibodies (ANA), anti-Ro/SS-A, anti-La/SS-B, anti-Sm, anti-ribonucleoprotein (anti-RNP), anti-smooth muscle antibodies (anti SMA), anti-parietal cell antibodies (anti-PCA), anti-liver-kidney microsome type 1 (anti-LKM-1) antibodies and anti-mitochondrial antibodies (AMA). ANA were detected in 278 (83%) patients. The association of positive ANA with the presence of anti-Ro/SS-A and anti-La/SS-B antibodies reached statistical significance at a titre of ANA >1/80 (p<0.001), while the presence of anti-Sm and anti-RNP was associated with positive ANA at a titre > or =1/320 (p=0.037 for Sm and p=0.016 for RNP). ANA titres correlated with the number of positive antibodies against specific nuclear antigens (p<0.001) but not with the number of positive antibodies against non-nuclear antigens. We found positive anti-Ro/SS-A antibodies in 111 (33%) patients, anti-La/SS-B in 78 (23%), anti-RNP in 8 (2%) and anti-Sm in 4 (1%). Anti-SMA antibodies were detected in 208 (62%) patients, with no significant associations with clinical or analytical SS features, while anti-PCA antibodies were found in 90 (27%) patients and were associated with a higher prevalence of thyroiditis and liver involvement. AMA were detected in 28 (8%) patients, although only 14 presented clinical and/or analytical evidence of liver involvement. No patient presented anti-LKM antibodies. ANA play a central role in the immunological expression of primary SS, due to their frequency and close association with the underlying presence of one or more anti-ENA antibodies. Positivity for antibodies against non-nuclear antigens such as anti PCA and AMA suggests an association with some organ-specific autoimmune diseases (thyroiditis and primary biliary cirrhosis), while the presence of anti-SMA, in spite of their high prevalence, has no clinical significance in primary SS. PMID- 16249827 TI - The symptomatic effects of intra-articular administration of hylan G-F 20 on osteoarthritis of the hip: clinical data of 6 months follow-up. AB - Hip osteoarthritis (OA) is usually managed with systemic treatments such as nonsteroidal anti-inflammatory drugs (NSAIDs) and/or symptomatic slow-acting drugs. Unfortunately, many patients either cannot tolerate NSAIDs or suffer serious, even fatal, NSAID-induced side effects, predominantly gastrointestinal ulceration and bleeding. Viscosupplementation, which aims to restore physiological and rheological features of the synovial fluid, is a well-accepted therapeutic option in knee OA patients, but limited data exist in the literature about its potential benefit for the treatment of hip OA. The purpose of this study is to observe the effects of hylan G-F 20 administered through ultrasound (US)-guided intra-articular (IA) injections in patients with symptomatic hip OA. We treated 30 patients with symptomatic hip OA. Under US guidance, 7 patients received one injection, 21 patients had two injections, and 2 patients received three injections, each with 2 ml of hylan G-F 20. Lequesne index, VAS scale of hip pain, and NSAID consumption were evaluated at baseline as well as 2 and 6 months after the beginning of the treatment. No systemic adverse events were observed. Lequesne index, VAS pain score, and NSAID consumption showed a reduction that was statistically significant to the baseline. The present observation suggests the potentiality for the safety and efficacy of hylan G-F 20 injected under US guidance in patients with symptomatic hip OA. Further controlled studies are needed. PMID- 16249828 TI - Severe rheumatoid valvular heart disease. AB - Rheumatoid arthritis (RA) is a systemic inflammatory disease in which extra articular involvement is not uncommon. Cardiac compromise may be frequent, although most often, it is clinically silent. Herein, two cases of RA-related endocarditis, one of which required valve replacement, are described. Etanercept was useful in controlling the articular and extra-articular RA compromise in both cases. PMID- 16249829 TI - A novel approach of local corticosteroid injection for the treatment of carpal tunnel syndrome. AB - The objective of the study was to compare the favorable response rate, time duration, and pain level of local corticosteroid injection using a novel approach for the treatment of carpal tunnel syndrome vs a classic approach. Patients with symptomatic carpal tunnel syndrome of less than 1-year duration were randomized for local corticosteroid injection using either the classic approach or a novel approach. In our approach (novel), we used a 29 gauge x 1/2-in. needle and a 1-ml insulin syringe containing 12 mg of methylprednisolone mixed with 0.15 ml of lidocaine 2%, and the site of the injection was 2-3 cm distal to the middle of wrist crease. In the classic approach, we used a 25 gauge x 3-cm needle and a 2 ml syringe injecting 35 mg of methylprednisolone mixed with 0.5 ml of lidocaine 2%, 3-4 cm proximal to the wrist crease and just ulnar to the tendon of the flexor carpi radialis muscle. Response rate was evaluated 1, 3, 6, and 12 weeks after the injection, and also the duration of time of the procedure and the level of pain using the visual analogue scale were compared between the two groups. Forty-two patients signed the consent form, and all of them completed the study [21 patients in the classic approach group (group 1) and 21 patients in the novel approach group (group 2)]. The favorable response rates were 100, 81, 71, and 57% in group 1 and 100, 71, 67, and 57% in group 2 after 1, 3, 6, and 12 weeks, respectively. There was no significant difference in the favorable response rate between the two groups (p=0.468, 95% CI=-12-31%, after 3 weeks). The average duration of time of the procedure in group 1 was 26.71+/-32.83 s compared to 8.48+/-1.123 s (p=0.021) in group 2. The average grade of pain expressed by the patients in group 1 was 4.38+/-1.523 compared to 3.62+/-1.071 in group 2 (p=0.065). In conclusion, local corticosteroid injection using the novel approach for the treatment of carpal tunnel syndrome is helpful, and the favorable response rates are comparable to those using the classic approach after 1, 3, 6, and 12 weeks. The novel approach is much less time consuming and is not more painful. PMID- 16249830 TI - Hypertrophic pachymeningitis and undifferentiated connective tissue disease: a case report and review of the literature. AB - A 63-year-old man with hypertrophic pachymeningitis (HP) and an undifferentiated connective tissue disease is described. Combined therapy with prednisolone and azathioprine improved his symptoms. The association between an undifferentiated connective tissue disease and HP is discussed. PMID- 16249831 TI - Lipid peroxidation and trace elements in systemic sclerosis. AB - Oxidative stress appears to be important in the causation and perpetuation of tissue injury and fibrosis in systemic sclerosis or scleroderma (SSc). We conducted a case-control study to assess lipid peroxidation levels as determined by measuring fasting plasma malondialdehyde (MDA) and serum levels of the trace elements selenium, iron, zinc and copper in SSc. Plasma MDA levels were almost tenfold higher in patients than in controls (p=0.00007), and an inverse relationship between MDA levels and disease duration (r=-0.52, p=0.044) was observed. Selenium levels were lower in patients than in controls (p=0.012). Within the patient cohort, copper correlated inversely with the total skin score (r=-0.52, p=0.03). Our findings provide further evidence that lipid peroxidation is increased and antioxidant capacity is reduced in SSc. The gradual decline in MDA levels with time suggests that antioxidant therapy, if to be useful in SSc, is most likely to be effective early in the course of the disease. PMID- 16249832 TI - The development of pelvic organ prolapse after colposuspension: a prospective, long-term follow-up study on the prevalence and predisposing factors. AB - The objectives of this prospective study were to determine the prevalence of pelvic organ prolapse (POP) after colposuspension and to investigate possible preoperative and operative risk factors. Seventy-seven women who underwent colposuspension between 1996 and 1997 were investigated. POP was assessed before colposuspension using the pelvic organ prolapse quantification system (POPQ). Women were reassessed at one and seven to eight years (or when referred with symptomatic POP). By seven to eight years, of the 77 women, 29 (38%) had developed symptomatic prolapse, 29 (38%) had asymptomatic prolapse, 7 (9%) had no symptoms and no prolapse, and 12 (15%) could not be assessed. POP at one year was significantly associated with the presence of posterior vaginal descent before colposuspension (odds ratio 3.07, 95% CI 1.10-8.60, p = 0.03). No variable reached statistical significance by eight years postcolposuspension. In conclusion, this is the first study to assess POP prospectively using a validated method before and after colposuspension. The results add support to the view that there is an association between colposuspension and the development of symptomatic POP (requiring surgery). PMID- 16249833 TI - Urinary stress incontinence treatment with Remeex in a patient with epispadias: a case report. AB - Epispadias is a rare condition in women. A postmenopausal 53-year-old woman with a known problem of epispadias came to our outpatient clinic complaining of persistent severe urinary stress incontinence without overactive bladder symptoms. She had previously undergone an inefficacious tension-free vaginal tape (TVT) procedure in another institution. We implanted a Remeex (Mechanic External Regulation) device without applying any tension to the sling. Two tape regulations were performed the day after surgery and 2 months later to gain continence. At 13 months of follow up, this woman was asymptomatic and satisfied. As documented by this case here, the Remeex procedure is particularly suitable in selected high-risk patients where TVT and TVT live slings are less effective. PMID- 16249834 TI - Similarity-based descriptors (SIBAR)--a tool for safe exchange of chemical information? AB - Exchange of chemical information without disclosure of the respective structures would greatly increase the data sets available for model building. Within the framework of the ChemMask project we explored the principal applicability of SIBAR-descriptors to mask chemical structures. SIBAR is based on calculation of similarity values for each compound of the training set to a set of reference compounds. Although the SIBAR-approach per se does not allow to unambiguously trace back the chemical structure of a compound, similarity searching in a 1.5 million compound database spiked with compounds structurally analogous to the query structure lead to the retrieval of compounds structurally and pharmacologically highly analogous to the "hidden" query structure in all three examples investigated. Comparison to results obtained with the original descriptors used to calculate the SIBAR-values showed, that SIBAR indeed adds some fuzziness to the data matrix. PMID- 16249835 TI - Flying blind: using a digital dashboard to navigate a complex PACS environment. AB - Radiology workflows have become more distributed and complicated, and fewer tangible cues are available to the radiologist to help optimize task prioritization and selection. Additionally, faster scanners, more detailed exams, and increased demand for imaging services have precipitated a potential image overload for today's radiologists who are pressured to provide efficient, quality service in less time. Radiologists are faced with the task of operating within complex systems but are lacking tools to efficiently and effectively monitor these systems in real time. Dashboard technology can help address this deficiency in radiology and facilitate informed, optimized decisions about workflow. Possible areas of application include workflow consolidation, workload distribution, and urgency evaluation. Dashboards should be optimized, context sensitive, customizable, and workflow-integrated. Further research is needed to identify the most important dashboard metrics, determine their optimal display, and validate their utility. PMID- 16249836 TI - ROC study of four LCD displays under typical medical center lighting conditions. AB - Nine observers reviewed a previously assembled library of 320 chest computed radiography (CR) images. Observers participated in four sessions, reading a different 1/4 of the sample on each of four liquid crystal displays: a 2 megapixel (MP) consumer color display, a 2-MP business color display, a 2-MP medical-grade gray display, and a 3-MP gray display. Each display was calibrated according to the DICOM Part 14 standard. The viewing application required observer log-in, then randomized the order of the subsample seen on the display, and timed the responses of the observer to render a 1-5 judgment on the absence or presence of ILD on chest CRs. Selections of 1-2 were considered negative, 3 was indeterminate, and 4-5 were positive. The order of viewing sessions was also randomized for each observer. The experiment was conducted under controlled lighting, temperature, and sound conditions to mimic conditions typically found in a patient examination room. Lighting was indirect, and illuminance at the display face was 195 +/- 8% lux and was monitored over the course of the experiment. The average observer sensitivity for the 2 MP color consumer, 2 MP business color, 2 MP gray, and 3 MP gray displays were 83.7%, 84.1%, 85.5%, and 86.7%, respectively. The only pairwise significant difference was between the 2 MP consumer color and the 2-MP gray (P = 0.05). Effect of order within a session was not significant (P = 0.21): period 1 (84.3%), period 2 (86.2%), period 3 (85.4%), period 4 (84.1%). Observer specificity for the various displays was not statistically significant (P = 0.21). Finally, a timing analysis showed no significant difference between the displays for the user group (P = 0.13), ranging from 5.3 s (2 MP color business) to 5.9 s (3 MP Gray). There was, however, a reduction in time over the study that was significant (P < 0.001) for all users; the group average decreased from 6.5 to 4.7 s per image. Physical measurements of the resolution, contrast, and noise properties of the displays were acquired. Most notably, the noise of the displays varied by 3.5x between the lowest and highest noise displays. Differences in display noise were indicative of observer performance. However, the large difference in the magnitude of the noise was not predictive of the small difference (3%) in the observer sensitivity for various displays. This is likely because detection of interstitial lung disease is limited by "anatomical noise" rather than display or x-ray image noise. PMID- 16249837 TI - Synchronized, interactive teleconferencing with digital cardiac images. AB - St James's Hospital is a tertiary referral center for percutaneous intervention and cardiothoracic surgery for a number of referring hospitals. This article reports on the development and implementation of a synchronized, interactive teleconferencing system for cardiac images that links St. James's Hospital with a remote site (Sligo General Hospital) and overcomes the problems of transmission of large image files. Teleconferencing was achieved by setting up lossless auto transmission of patient files overnight and conferencing the next morning with linked control signals and databases. As a suitable product was not available, a commercially new software was developed. The system links the imaging databases, monitors and synchronizes progress through imaging sequences, and links a range of image processing and control functions. All parties to the conference are ensured that they are looking at the same images as they are played or at specific aspects of an image that the other party is highlighting. The system allows patient management decisions to be made at a weekly joint teleconference with cardiothoracic surgeons and interventional cardiologists from both sites. Rapid decision making was facilitated with 70% of decisions obtained within 24 h, and 88% within 1 week of their procedure. In urgent cases, data can be transmitted within 20 min of the diagnostic procedure. The system allows increased access to angiography for patients living in rural areas, and provides a more focused referral for revascularization. Participation of the referring cardiologist has improved the quality of decision making. PMID- 16249838 TI - Defining the PACS profession: an initial survey of skills, training, and capabilities for PACS administrators. AB - The need for specialized individuals to manage picture archiving and communications systems (PACS) has been recognized with the creation of a new professional title: PACS administrator. This position requires skill sets that bridge the current domains of radiology technologists (RTs), information systems analysts, and radiology administrators. Health care organizations, however, have reported difficulty in defining the functions that a PACS administrator should perform-a challenge compounded when the tries to combine this complex set of capabilities into one individual. As part of a larger effort to define the PACS professional, we developed an extensive but not exclusive consensus list of business, technical, and behavioral competencies desirable in the dedicated PACS professional. Through an on-line survey, radiologists, RTs, information technology specialists, corporate information officers, and radiology administrators rated the importance of these competencies. The results of this survey are presented, and the implications for implementation in training and certification efforts are discussed. PMID- 16249839 TI - Disclosing the diagnosis of multiple sclerosis. AB - CONTEXT: The question of how best to disclose to patients the diagnosis of serious and/or incurable neurological diseases has been much explored, but that of when has received little rigorous study. The present study investigates this question in relation to multiple sclerosis (MS), a disease marked by its incurability, unpredictability and predilection for young adults. OBJECTIVES: We aimed to ascertain the preferences of Greek MS sufferers concerning when they should ideally be informed they have the disease, and their preferences and reactions regarding disclosure of the diagnosis. Design, setting and patients 1,200 Greek MS patients,members of the MS Society, were asked to complete a questionnaire regarding their experience of and attitudes towards receiving the diagnosis. DESIGN, SETTING AND PATIENTS: 1,200 Greek MS patients,members of the MS Society, were asked to complete a questionnaire regarding their experience of and attitudes towards receiving the diagnosis. RESULTS: 657 patients (55 %) responded. 91% favoured learning the diagnosis immediately, but only 44% had had this experience: 29% had been informed within 1-3 years, and 27% later. Interestingly, however, a significant minority (9 %) suggested a possible preference for delayed delivery of diagnosis and 23.2% stated that concealing the diagnosis would not lead to loss of confidence in their doctor. CONCLUSION: This study-the largest of its kind-provides objective data supporting prompt disclosure of diagnosis as the clearly-expressed preference amongst most patients. Interestingly, however, the results also re-emphasise the importance of a difficult medical art: attempting to judge whether an individual patient is one (of the 91%) preferring immediate disclosure-or of the nearly 1-in-10 (9%) who may not. PMID- 16249840 TI - Relationship between bone mineral density and myocardial infarction in US adults. AB - Cardiovascular disease and osteoporosis have several common risk factors, and quite a few studies suggest a relationship between them. The objective of the present study was to explore the relationship between cardiovascular disease risk factors and bone mineral density in association with having had a previous myocardial infarction in a general population. This cross-sectional study was conducted using data for 5,050 women and men aged 50-79 years who participated in the Third National Health and Nutrition Examination Survey (NHANES III). Race/ethnic and gender-specific mean BMD values for young adults were used to determine race/ethnic and gender-specific T-scores to define osteoporosis and low BMD. Multiple logistic regression analysis revealed that subjects self-reporting a previous myocardial infarction had significantly higher odds (odds ratio 1.28, [95% confidence interval (CI), 1.01 to 1.63] p=0.04) of having low bone mineral density, when adjusting for cardiovascular disease and osteoporosis risk factors. Self-reported myocardial infarction was not significantly associated with low bone mineral density in women, (odds ratio 1.22, [95% CI, 0.80 to 1.86] p=0.37), but was significant in men, (odds ratio 1.39, [95% CI, 1.03 to 1.87] p=0.03). These findings demonstrate that male survivors of myocardial infarction have low bone mineral density. The pathophysiologic connection between the atherosclerotic and the osteoporotic processes needs further elucidation. It is also of importance to study the processes in both men and women. PMID- 16249841 TI - Results of surgery for toxic multinodular goiter. AB - PURPOSE: We analyzed the clinical and histological features of patients operated on for toxic multinodular goiter (TMG) to determine the clinical profile and evaluate the surgical results. METHODS: We reviewed 672 patients who underwent surgery for multinodular goiter (MG), 112 (17%) of whom had hyperthyroidism, and analyzed the epidemiological, clinical, and surgical variables. RESULTS: The patients with TMG tended to be older than those with nontoxic MG, with a greater evolution time of the goiter and a higher rate of positive antithyroid antibodies. In the multivariate analysis, the only feature characteristic of TMG, as opposed to nontoxic MG, was the evolution time. Morbidity was 34%, representative of the fact that that most of the patients were seen before the establishment of our endocrine surgical unit. The hyperthyroid symptoms resolved in all patients, but 4 of 17 patients who underwent partial surgical resection showed signs of relapse within a follow-up period of 98 +/- 71 months. CONCLUSIONS: TMG is characterized by a long evolution time and is most effectively treated by total thyroidectomy, which achieves complete remission from symptoms, without relapse, and is necessary if there is associated carcinoma. However, the incidence of complications may be high if this procedure is not carried out by surgeons with experience in endocrine surgery. PMID- 16249842 TI - Prognostic factors in breast cancer: the value of the Nottingham Prognostic Index for patients treated in a single institution. AB - PURPOSE: The Nottingham Prognostic Index (NPI) is used to predict survival in patients with breast cancer. This index is based on tumor size, lymph node stage, and histological grade and allows the stratification of patients into three different prognostic groups. Our aim was to verify the effect of some prognostic variables on survival and to establish the independent influence of each of these variables by a survival regression analysis. We applied the NPI to the same group of patients to assess its predictive power and reproducibility. METHODS: We evaluated 311 women with breast cancer treated between January 1993 and December 1998. RESULTS: In a multivariate analysis (Cox proportional hazard model), only size, lymph node involvement, and histological grade were independent prognostic factors. The survival curves obtained after applying the NPI were similar to those for the factors with independent prognostic significance derived from our multivariate analysis. CONCLUSION: The NPI allows us to accurately predict prognosis, and we advocate its standardized use. PMID- 16249843 TI - Group IIA-soluble phospholipase A2 levels in patients with infections after esophageal cancer surgery. AB - PURPOSE: To examine the changes in blood-soluble phospholipase A(2)-IIA levels caused by surgical stress and postoperative infections. METHODS: We retrospectively analyzed a prospective database of 40 patients who underwent esophagectomy for esophageal cancer. Nine of these patients had a postoperative infection (E Inf(+) group), and 31 did not have a postoperative infection (E Inf( ) group). The blood sPLA(2)-IIA level was measured using a radioimmunoassay, and whole blood was stimulated with lipopolysaccharide (LPS) to examine the sPLA(2) IIA production. RESULTS: In the E Inf(-) group, the blood sPLA(2)-IIA levels peaked on postoperative day (POD) 3 then decreased gradually thereafter. Receiver operator characteristic statistics based on the sPLA(2)-IIA values on POD 5, which are used to classify postoperative infectious complications, revealed an area under the curve of 0.789. However, stimulation of peripheral blood cells with LPS did not induce the production of sPLA(2)-IIA. CONCLUSION: During the early postoperative phase, blood sPLA(2)-IIA levels increase according to the surgical stress. Soluble PLA(2)-IIA may be produced at the site of infection or in the liver, but not in the circulating blood. Sustained elevation of the serum sPLA(2)-IIA level, observed even after POD 3, seems to represent a response to postoperative infection. PMID- 16249844 TI - Peritoneal carcinoembryonic antigen level for predicting locoregional and distant spread of gastric cancer. AB - PURPOSE: Peritoneal recurrence is not an uncommon cause of death after surgery for gastric cancer, even after surgery with curative intent. This indicates that there is undetected residual disease in the peritoneal cavity. We conducted this study to determine the value of peritoneal and serum carcinoembryonic antigen (CEA) levels and peritoneal washing cytology in predicting the locoregional and distant spread of gastric cancer. METHODS: We prospectively evaluated 70 consecutive patients with gastric cancer by measuring peritoneal CEA (pCEA) and serum CEA (sCEA) levels and peritoneal washing cytology results, and studying their effect on the histopathologic properties. The effect of the pCEA level on disease-free survival (DFS) and overall survival (OS) was also evaluated in patients treated with curative intent. RESULTS: Twenty-one (30%) patients had sCEA levels >10 ng/ml, whereas 25 patients (35.7%) had pCEA levels >10 ng/g protein and 26 patients (37.1%) had positive cytology. The pCEA levels were significantly higher in patients with hepatic metastases (P = 0.034), or serosal (P = 0.028), and peritoneal (P = 0.026) involvement, whereas the sCEA levels were significantly higher only in patients with hepatic metastases (P = 0.04). Similarly, positive cytology was mainly detected in patients with hepatic metastases (P = 0.004). The pCEA levels significantly affected DFS (P = 0.002) and OS (P = 0.01) in 34 patients treated with curative intent. CONCLUSION: Since pCEA levels are more useful for predicting locoregional recurrence, their measurement during surgery may help plan the most appropriate surgical strategy and adjuvant therapy. PMID- 16249845 TI - Laparoscopic cholecystocholangiography as an effective alternative exploratory laparotomy for the differentiation of biliary atresia. AB - PURPOSE: Exploratory laparotomy with cholecystocholangiography is often performed for the definitive diagnosis of biliary atresia (BA) when radiological and biochemical studies are inconclusive. Laparoscopic cholecystocholangiography (LCC) has recently been introduced as an alternative procedure to avoid unnecessary laparotomy. We discuss the advantages and indications of LCC based on our experience of performing this diagnostic procedure in four infants with prolonged direct hyperbilirubinemia. METHODS: We performed LCC in four infants by direct percutaneous puncture of the gallbladder by inserting a cholangiocatheter into the gallbladder or cystic duct. The decision to perform LCC was based on ultrasonography and computed tomography findings of an atrophic gallbladder with a patent lumen, although analysis of duodenal juice and hepatobiliary scintigraphy showed no sign of the excretion of bile into the small intestine. RESULTS: In three infants, LCC did not show the entire biliary system, and laparotomy was necessary to confirm the diagnosis of BA. However, in the other patient LCC showed a normal biliary system and BA was excluded. CONCLUSION: Laparoscopic cholecystocholangiography may be useful for the differential diagnosis of BA in patients with a patent gallbladder when imaging and biochemical findings are inconclusive. Furthermore, unnecessary exploratory laparotomy may be avoided in patients without BA. PMID- 16249846 TI - Anterolateral thoracotomy for distal aortic arch disease. AB - PURPOSE: Optimal exposure and antegrade arterial perfusion are keys to avoiding complications in the repair of distal aortic arch disease. To achieve these ends, we performed distal aortic arch repair through a left anterolateral thoracotomy while also using axillary artery perfusion. METHODS: From Mach 1998 to December 2004, 28 patients (23 men and 5 women, age 65.2 +/- 12.0 years) underwent a distal aortic arch repair through a left anterolateral thoracotomy. All cases had atherosclerotic aneurysms. Emergency surgery was performed in 1 of these cases (1/28, 3.6%) with an aortic rupture. The right axillary artery was used for arterial perfusion in all cases. RESULTS: No perfusion problems occurred during surgery, and the left anterolateral thoracotomy approach provided an excellent view of both the aortic arch and descending aorta. There was no hospital mortality. Morbidity included one incident of transient convulsion without computed tomographic evidence of an embolism and one incident of heart failure that required temporary mechanical support. No other significant event or morbidity occurred related to the surgical methods. There was no late death during the 1 to 81-month follow-up. CONCLUSION: A left anterolateral thoractomy provides an ideal view of distal aortic arch disease, and antegrade arterial perfusion is effective in the prevention of retrograde embolism. These results suggest this treatment modality to be a reliable alternative approach for the repair of distal aortic arch disease. PMID- 16249847 TI - Diagnostic peritoneal lavage for diagnosing blunt hollow visceral injury: the accuracy of two different criteria and their combination. AB - PURPOSE: To test the usefulness of diagnostic peritoneal lavage (DPL) for identifying blunt hollow visceral injury with two different sets of criteria or a combination of the two. METHODS: Fifty victims with physical examinations and/or computed tomography findings equivocal for blunt hollow visceral injury underwent DPL. Whether or not to perform surgery was determined based on Otomo's DPL criteria [lavage white blood cell counts (L-WBC) over lavage red blood cell counts (L-RBC) divided by 150 (L-WBC > or = L-RBC/150) in the presence of hemoperitoneum, or L-WBC over 500/mm(3) (L-WBC > or = 500) in the absence of hemoperitoneum]. The cell count ratio, a comparison of L-WBC, L-RBC, peripheral WBC (P-WBC), and peripheral RBC (P-RBC) [(L-WBC/L-RBC)/(P-WBC/P-RBC) > or = 1] were all calculated retrospectively. RESULTS: There were one and two false positive cases based on Otomo's criteria and the cell count ratio, respectively, with corresponding accuracies of 97.8% and 95.7%, respectively. There were no false-positive or -negative cases according to the combined use of Otomo's criteria and cell count ratio, yielding an accuracy of 100%. CONCLUSION: Although each criterion alone is very accurate in predicting the presence of blunt hollow visceral injury, the combined use of the two would further improve the accuracy of the diagnosis and thereby reduce the number of unnecessary celiotomies. PMID- 16249848 TI - Bioresorbable hyaluronate-carboxymethylcellulose membrane (Seprafilm) in surgery for rectal carcinoma: a prospective randomized clinical trial. AB - PURPOSE: To evaluate the effectiveness of Seprafilm in preventing abdominal adhesions after radical resection of rectal carcinoma, and to observe whether Seprafilm had any adverse effects in patients treated with radiotherapy and chemotherapy. METHODS: A total of 62 patients participated in this prospective randomized clinical study, which was conducted to compare the outcomes of patients operated on with Seprafilm (SEPRA+) with those operated on without Seprafilm (SEPRA-). All patients received preoperative radiotherapy, followed by a two-stage operation, and 5-fluorouracil (5-FU)-based systemic chemotherapy. The primary endpoint of severity and extent of adhesions were evaluated at the time of ileostomy closure. The secondary endpoint included the recurrence of tumors, late complications, and outcome. RESULTS: Seprafilm significantly reduced the adhesions in both the midline incision area and peristomal area. This in turn reduced the operation time, blood loss, and extent of the incision at ileostomy closure. Seprafilm was not associated with any postoperative complications or chemoradiation-related toxicity, nor did it affect recurrence or survival rates. CONCLUSION: Seprafilm effectively reduced abdominal adhesions in chemoradiated patients, and had no adverse effects on the oncologic results of fully introduced adjuvant therapy. Thus, Seprafilm is a safe and effective tool for use in rectal carcinoma surgery. PMID- 16249849 TI - An assessment of the effects of two types of bioresorbable barriers to prevent postoperative intra-abdominal adhesions in rats. AB - PURPOSE: This study assessed and compared the efficacy of two types of bioresorbable membranes in the prevention of postoperative adhesion under clean contaminated and bacterial peritonitis conditions using a cecal ligation and puncture model in rats. METHODS: Wistar albino rats (n = 72) were divided into six groups. Bacterial peritonitis was induced using a cecal ligation and puncture model in groups 2, 4, and 6. Groups 1, 3, and 5 served as controls for clean contaminated procedures in the absence of bacterial peritonitis. Groups 1 and 2 were the untreated clean contaminated and bacterial peritonitis groups and served as controls for the effect of the bioresorbable membranes in each condition. In groups 3 and 4, a 1.5 x 3 cm USP glycerol/sodium hyaluronate/carboxymethylcellulose membrane was wrapped around the cecal resection area and a 2 x 4 cm membrane was left under the incision. The oxidized regenerated cellulose membrane was similarly applied in groups 5 and 6. Four weeks later, the adhesions were evaluated. In addition, fibrosis and inflammation were observed histopathologically. RESULTS: Adhesion development (P = .008), fibrosis (P = .008), and inflammation (P = .0001) differed among the groups. Both materials increased adhesion formation in the bacterial peritonitis condition. Increased fibrotic activity was detected in all material-applied groups under both conditions. In addition, more inflammation was detected in the groups that received the application of a material, especially in the presence of bacterial peritonitis. CONCLUSION: Neither material prevented adhesions in clean contaminated conditions. Moreover, they increased adhesion formation in bacterial peritonitis. PMID- 16249850 TI - A subacute hypoxic model using a pig. AB - PURPOSE: A large animal model of hypoxia is necessary to develop a new therapeutic method for respiratory failure. METHODS: The experiments were performed on six pigs weighing from 15 to 19 kg. Under general anesthesia the left chest was opened and the left main bronchus was closed by a stapler. A Swan Ganz catheter was inserted through the right jugular vein. The right carotid artery was cannulated and the mean arterial blood pressure was monitored, and arterial blood samples were drawn every 24 h until 96 h after the operation. The blood samples were submitted for a blood gas analysis. All data were expressed as the mean +/- standard deviation of the mean. RESULTS: The partial pressure of the oxygen of the arterial blood at baseline (104.8 +/- 24.3 mmHg) significantly decreased at 24 h after closure of the bronchus (76.7 +/- 9.9 mmHg, P < 0.001) and thereafter remained at the same level for 4 days. CONCLUSION: This hypoxic model using a pig was found to be very simple, effective, and reproducible. This model can be used for a variety of experiments to evaluate new therapeutic modalities for respiratory failure. PMID- 16249853 TI - Postchemotherapy resection of a primary mature malignant retroperitoneal teratoma in an adult: report of a case. AB - We herein report a rare case of a 47-year-old man with a primary mature retroperitoneal malignant teratoma. The patient received neoadjuvant chemotherapy and the residual tumor was resected. A histopathological examination confirmed the diagnosis of malignant teratoma with endodermal, ectodermal, and mesenchymal structures. The patient is alive and disease free at 13 months after resection. PMID- 16249851 TI - Albumin-producing hepatocytes derived from cryopreserved F344 rat bone marrow cells transplanted in the livers of congenic Nagase's analbuminemic rats. AB - PURPOSE: Hematopoietic stem cells (SCs) are thought to have the potential to differentiate into hepatocytes; however, this potential has not been reported for cryopreserved SCs. We investigated whether cryopreserved bone marrow cells (BMCs) from F344 rats (F344) can induce the growth of albumin-producing hepatocytes in the livers of congenic Nagase's analbuminemic rats (F344alb). METHODS: F344 BMCs were cryopreserved in University of Wisconsin (UW) solution containing 10% fetal bovine serum and 12% dimethylsulfoxide, at -80 degrees C. After thawing, 20 x 10(6) cells were infused via the portal vein into the livers of F344alb immediately after 70% hepatectomy (PH). We examined the recipient livers for albumin-positive (alb+) hepatocytes and albumin mRNA, and measured the serum albumin levels 4 weeks later. RESULTS: Single and double alb+ hepatocytes were occasionally seen in the F344alb livers without the BMC transplantation. However, clusters consisting of more than three alb+ hepatocytes were seen in the livers of recipients transplanted with the cryopreserved BMCs after PH, the same as in the livers transplanted with freshly isolated BMCs. Normal albumin mRNA was detected in the recipient livers and the serum albumin levels were increased. CONCLUSION: Cryopreserved F344 BMCs can induce the growth of alb+ hepatocytes after transplantion in the F344alb liver after PH. PMID- 16249852 TI - Mesenteric inflammatory veno-occlusive disease as a cause of acute abdomen: report of five cases. AB - Mesenteric inflammatory veno-occlusive disease (MIVOD) is a rare but increasingly recognized cause of intestinal ischemia. It can be defined as phlebitis or venulitis affecting either the bowel or mesentery, without any evidence of coexisting arterial inflammatory involvement or an obvious predisposing cause. We report the clinicopathological characteristics of five patients who, after presenting with an acute abdomen, underwent exploratory laparotomy and resection of ischemic bowel. The distinctive histopathological characteristics of MIVOD were identified in all five patients. We review the literature on this under reported condition. PMID- 16249854 TI - Pleomorphic carcinoma of the lung with mediastinal extension following malignant lymphoma: report of a case. AB - In following up a patient with non-Hodgkin's lymphoma, we encountered a case of pulmonary pleomorphic carcinoma with mediastinal direct invasion. A 65-year-old man with hemoptysis was found to have an abnormal shadow in the right upper lung field. A 6.4 x 4.8-cm tumor adjacent to the upper mediastinum occupied the right anterior segment of the upper lobe (S3) and invaded the superior vena cava (SVC). The serum level of neuron-specific enolase was elevated to 11.9 ng/ml. A specimen from a transbronchial lung biopsy of the right B3b bronchus revealed giant tumor cells. A right upper lobectomy with SVC reconstruction was performed. The resected tumor was diagnosed as a pulmonary pleomorphic carcinoma with a large component of giant and spindle cells, and it is considered to be a rare histologic type. PMID- 16249855 TI - Pulmonary tractotomy for a patient with traumatic penetrating lung injury: report of a case. AB - We report a case of traumatic hemopneumothorax caused by penetrating lung injury in a 26-year-old man. The patient underwent emergency thoractomy, which revealed hemorrhage in the lingular segment of the left lung. We found the bleeding point and controlled the hemorrhage using pulmonary tractotomy by inserting a linear stapler into the stab wound in the pulmonary parenchyma. The original technique of pulmonary tractotomy was performed for complete through-and-through injury by dividing the bridge of lung tissue between the aortic clamps. We were able to apply this procedure safely to stop bleeding from a stab wound that did not go through the lung. Thus, pulmonary tractotomy is an effective damage-control operation for the lung with obvious advantages over major lung resection. PMID- 16249856 TI - Resection of a pulmonary lesion after liver transplantation: report of a case. AB - A 44-year-old Chinese-Indonesian man who underwent living-donor liver transplantation with a right liver graft presented 4 months later with a cough and fever. Chest X-ray showed a nodular shadow in the apex of the left lung, which was diagnosed as pulmonary tuberculosis. After 1 week of antituberculous chemotherapy, we performed a left upper lobectomy. Postoperative antituberculous chemotherapy, consisting of isoniazid (300 mg/day) and rifampin (450 mg/day), was continued for 4 months, and there has been no sign of recurrence for 1 year since the thoracotomy. This case supports the feasibility of surgery for localized pulmonary tuberculosis soon after transplantation. PMID- 16249857 TI - True solitary pancreatic cyst in an adult: report of a case. AB - The differential diagnosis of cystic neoformations in the pancreas is challenging. We report a case of a true solitary cyst of the pancreas in a 26 year old woman. Abdominal magnetic resonance imaging and computed tomography showed a unilocular neoformation in the head of the pancreas, without obstruction of Wirsung's duct. We excised the cyst and performed Roux-en-Y loop pancreaticojejunostomy, but the patient suffered recurrent acute pancreatitis from Wirsung's duct stenosis. Thus, a new Roux-en-Y loop pancreaticojejunostomy was successfully done 6 months later. Histologically, the cyst was lined by cuboidal epithelium, immunohistochemically positive to anti-carbohydrate antigen 19-9 antibodies. To our knowledge, only 11 cases of solitary true cyst of the pancreas in adults have been reported, so the characteristics of this unusual entity are not well known. We propose a scheme for the differential diagnosis of cystic neoformations of the pancreas, starting from the histopathological definition of a true solitary cyst. PMID- 16249858 TI - Disposal of replaced common hepatic artery coursing within the pancreas during pancreatoduodenectomy: report of a case. AB - A replaced common hepatic artery (RCHA) originating from the superior mesenteric artery (SMA) is a rare anomaly. We herein report such a case in a 62-year-old man who was scheduled to undergo a pancreatoduodenectomy for lower bile duct cancer. Computed tomography (CT) showed the RCHA to run along the ventral side of the pancreas. Abdominal angiography showed an RCHA originating from the SMA, which communicated with an aberrant left hepatic artery from the left gastric artery. No gastroduodenal artery was observed, but instead a direct ramification of a right gastroepiploic artery was seen. Similar cases from the English literature were reviewed. The RCHA was confirmed to course first along the ventral side of, and then within, the pancreas. Clamping of the RCHA did not influence the arterial flow in the liver, and the RCHA was subsequently divided without reconstruction. In three of the five reviewed cases in which the RCHA coursed either within or along the ventral side of the pancreas, no gastroduodenal artery was found, but instead a direct ramification of a right gastroepiploic artery was observed. A combination of CT and angiographic findings can help in both the diagnosis of an anomalous RCHA coursing either within or along the ventral side of the pancreas as well as in selecting optimal operative procedures. Pancreatoduodenectomy was performed with a curative resection according to our usual practice except for the fact that we preserved the aberrant left hepatic artery. PMID- 16249859 TI - Splenic metastasis from esophageal cancer: report of a case. AB - The spleen is an unusual site of metastasis from an esophageal malignancy. We herein report the case of a 25-year-old woman who underwent a transhiatal esophagectomy and adjuvant radiotherapy and chemotherapy for squamous cell carcinoma of the lower third of the esophagus with pN1 lymph node metastasis. Fifteen months following surgery she was found to have splenic metastasis infiltrating the tail of the pancreas at the hilum. A splenectomy, distal pancreatectomy, and resection of the splenic flexure with colocolic anastomosis were performed. A histological examination of the resected specimen showed squamous cell carcinoma. PMID- 16249860 TI - Successful surgical treatment of an infrarenal abdominal pseudoaneurysm caused by tuberculosis: report of a case. AB - A 76-year-old man was admitted to our hospital for investigation of an apparent abdominal aortic aneurysm detected during treatment for epididymitis. A chest X ray showed miliary shadows in the bilateral lung fields strongly suggestive of tuberculosis. The diameter of the aneurysm increased, and examinations showed impending rupture of a pseudoaneurysm. However, a definitive disease pathogenesis was not obtained before surgery. We performed a subemergency operation, which revealed an infrarenal abdominal pseudoaneurysm caused by tuberculosis. The pseudoaneurysm appeared to have resulted from direct extension of tuberculous lymphadenitis to the aortic wall, which ruptured. We review 24 other cases of tuberculous aortic aneurysms surgically treated in Japan before 2004. PMID- 16249861 TI - Late complications following surgery for type A acute aortic dissection using gelatin-resorcin-formaldehyde glue: report of two cases. AB - Gelatin-resorcin-formaldehyde (GRF) glue has been widely applied during operations for acute aortic dissection (AAD). At our institution, GRF glue was applied in 40 patients who underwent surgical procedures for AAD from 1995, two of whom needed a reoperation because of the development of a redissection and/or a pseudoaneurysm at the anastomotic sites. The operative findings and histological examinations suggested that the application of GRF glue during the initial operations might be related to the development of a redissection and/or a pseudoaneurysm. PMID- 16249862 TI - Aortotomy and endarterectomy of the ascending aorta for aortic valve replacement in a patient with porcelain aorta. AB - Patients with a calcified "porcelain" aorta may also have aortic valve stenosis, necessitating aortic valve replacement to prevent cerebral complications. However, a porcelain aorta can be difficult to open, and ascending aorta repair sutures are almost impossible to insert in a calcified plate. We devised a method of aortotomy using a small oscillating saw, which allowed us to incise the aortic wall cleanly without destruction or the formation of debris. After partial endarterectomy of the calcified plate with aortotomy, closure of the aorta was buttressed with bovine pericardium. During the endarterectomy, we used an elevator designed for hand surgery to exfoliate the calcified intimal plate. PMID- 16249863 TI - Congenital diseases of the thoracic aorta. Role of MRI and MRA. AB - Aortic malformations may be associated with other congenital heart abnormalities or may present independently, as incidental findings in asymptomatic patients. For more than 30 years, conventional imaging techniques for detection and assessment of congenital anomalies of the aorta have been chest X-ray, echocardiography and angiography. In recent times, considerable interest in congenital aortic diseases has been shown, due to technical progresses of noninvasive imaging modalities. Among them, magnetic resonance imaging (MRI) almost certainly offers the greatest advantages, especially in young patients in which a radiation exposure must be avoided as much as possible. MRI provides an excellent visualization of vascular structures with a wide field of view, well suited for evaluation of the thoracic aorta malformations. With the implementation of magnetic resonance angiography (MRA) it is also possible to depict any relationship with supra-aortic or mediastinal vessels. Phase contrast technique allows identification of the hemodynamic significance of the aortic alteration. Some technical considerations, which include fast spin-echo, gradient echo and, especially, MRA techniques with phase-contrast and contrast enhanced methods, are discussed and applied in the evaluation of congenital thoracic aorta diseases. PMID- 16249864 TI - Detection of pericardial inflammation with late-enhancement cardiac magnetic resonance imaging: initial results. AB - OBJECTIVE: To examine the value of late-enhancement cardiac magnetic resonance imaging (MRI) for detection of pericardial inflammation. MATERIALS AND METHODS: Late-enhancement cardiac MRI was performed in 16 patients with clinical suspicion of pericardial disease. Pericardial effusion, pericardial thickening and pericardial enhancement were assessed. MRI findings were compared with those of definitive pericardial histology (n=14) or microbiology (n=2). A control group of 12 patients with no clinical evidence of pericardial disease were also imaged with the same MRI protocol. RESULTS: Sensitivity and specificity for late enhancement MRI detection of pericardial inflammation was of 100%. There was MRI late enhancement of the pericardial layers in all five patients with histological/microbiological evidence of inflammatory pericarditis. MRI demonstrated no pericardial thickening and no MRI late enhancement with or without a pericardial effusion in any of the five patients with histological evidence of a normal pericardium. MRI detected pericardial thickening in the absence of both pericardial effusion and late enhancement in all six patients with histological evidence of chronic fibrosing pericarditis. The 12 control subjects showed no evidence of pericardial MRI late enhancement. CONCLUSIONS: These findings demonstrate that MRI late enhancement can be used to visualize pericardial inflammation in patients with clinical suspicion of pericardial disease. PMID- 16249865 TI - The value of magnetic resonance imaging for the diagnosis of arrhythmogenic right ventricular cardiomyopathy. AB - This study evaluated the diagnostic significance of a magnetic resonance imaging (MRI) based scoring model for identification of arrhythmogenic right ventricular cardiomyopathy (ARVC) in patients with MRI evidence of RV abnormalities. Fifty three patients with RV myocardial abnormalities on MRI were divided into a group with ARVC 1 (n=17) and a group with other RV arrhythmias (n=37). Decision tree learning (DTL) and linear classification (based on a modified ARVC scoring model of major and minor criteria) were used to identify and assess MRI criterion information value, and to induce ARVC diagnostic rules. All major ARVC criteria were more frequent in the ARVC group. Among minor criteria regional RV hypokinesia, mild segmental RV dilatation, and prominent trabeculae were more frequent in the ARVC group while mild global RV dilatation was more frequent in the non-ARVC group. RV aneurysm achieved highest importance in ARVC diagnosis (predictive accuracy 76.8%). Better diagnostic accuracy (sensitivity 93.3%, specificity 89.5%) was achieved when the MRI score for the major and minor criteria reached threshold value of four: two major criteria, or one major and two minor, or four minor criteria. Combinations between major and minor criteria contributed to a statistically valid model for ARVC diagnosis. PMID- 16249866 TI - Functional clusters in the human parietal cortex as revealed by an observer independent meta-analysis of functional activation studies. AB - The human parietal cortex is a highly differentiated structure consisting of cytoarchitectonically defined subareas that are specifically connected with other cortical and subcortical areas. Based on evidence from neurophysiological studies in subhuman primates these subareas are supposed to be functionally highly specialized. Here, we reviewed 51 different neuroimaging studies on healthy subjects with activation of the parietal lobe in statistical parametric maps. Running a cluster analysis on the stereotactic coordinates of the centers of gravity of the activation areas and plotting them into Talairach space showed a high consistency of the mean activation foci for similar paradigms across different laboratories and functional imaging modalities. Our meta-analysis exposed seven distinct pairs of quite symmetrically distributed subareas of the parietal cortex of each hemisphere as well as three unpaired regions that are critically involved in the generation of limb and eye movements in egocentric and allocentric coordinates, but also in attention, memory and cognitive problem solving. These data highlights the modular organization of the human parietal lobe. By its locally interspersed distributed circuits it orchestrates specialized cognitive subfunctions interfacing perception and action. Our meta analysis provides a new framework for understanding information processing in the human parietal cortex. PMID- 16249867 TI - Quantitative architectural analysis: a new approach to cortical mapping. AB - Recent progress in anatomical and functional MRI has revived the demand for a reliable, topographic map of the human cerebral cortex. Till date, interpretations of specific activations found in functional imaging studies and their topographical analysis in a spatial reference system are, often, still based on classical architectonic maps. The most commonly used reference atlas is that of Brodmann and his successors, despite its severe inherent drawbacks. One obvious weakness in traditional, architectural mapping is the subjective nature of localising borders between cortical areas, by means of a purely visual, microscopical examination of histological specimens. To overcome this limitation, more objective, quantitative mapping procedures have been established in the past years. The quantification of the neocortical, laminar pattern by defining intensity line profiles across the cortical layers, has a long tradition. During the last years, this method has been extended to enable a reliable, reproducible mapping of the cortex based on image analysis and multivariate statistics. Methodological approaches to such algorithm-based, cortical mapping were published for various architectural modalities. In our contribution, principles of algorithm-based mapping are described for cyto- and receptorarchitecture. In a cytoarchitectural parcellation of the human auditory cortex, using a sliding window procedure, the classical areal pattern of the human superior temporal gyrus was modified by a replacing of Brodmann's areas 41, 42, 22 and parts of area 21, with a novel, more detailed map. An extension and optimisation of the sliding window procedure to the specific requirements of receptorarchitectonic mapping, is also described using the macaque central sulcus and adjacent superior parietal lobule as a second, biologically independent example. Algorithm-based mapping procedures, however, are not limited to these two architectural modalities, but can be applied to all images in which a laminar cortical pattern can be detected and quantified, e.g. myeloarchitectonic and in vivo high resolution MR imaging. Defining cortical borders, based on changes in cortical lamination in high resolution, in vivo structural MR images will result in a rapid increase of our knowledge on the structural parcellation of the human cerebral cortex. PMID- 16249868 TI - [Infection epidemiological data among blood donors in Germany 2003-2004. Report of the Robert Koch Institute in accordance with Article 22 of the Transfusion Act]. AB - The Robert Koch Institute collects and evaluates data on the prevalence and incidence of HIV, hepatitis C (HCV), hepatitis B (HBV) and syphilis infections among blood and plasma donors in Germany in accordance with Article 22 of the Transfusion Act. This surveillance permits an assessment of the occurrence of infections in the blood donor population and consequently the safety of the collected donations. This report includes data from all blood donation services in Germany for 2003 and 2004. Altogether 7.09 million and 6.37 million donations or blood samples from prospective donors were screened in 2003 and 2004, respectively. In 2003 the prevalence rates of the relevant infections (per 100,000 donations) were 8.2 for HIV, 99.3 for HCV, 158.9 for HBV and 34.4 for syphilis. In 2003 the rates of seroconversion (per 100,000 donations) were 0.8 for HIV, 1.2 for HCV, 1.0 for HBV and 1.5 for syphilis in 2003. In 2004 the prevalence rates (per 100,000 donations) were 4.8 for HIV, 85.3 for HCV, 156.3 for HBV and 36.8 for syphilis. In 2004 the rates of seroconversion (per 100,000 donations) were 0.9 for HIV, 1.3 for HCV, 0.6 for HBV and 2.0 for syphilis. The analysis showed a very low incidence of infections with a decreasing trend for HCV infections and an increase in HIV and syphilis infections. The latter needs to be monitored carefully and possible causes evaluated. The quality of the reported data has improved compared to previous years. Still, some problems remain with the differentiation of the data according to sex, age and interdonation interval as well as reporting by individual blood donation centres as required by the Transfusion Act. PMID- 16249869 TI - Improved cotyledonary node method using an alternative explant derived from mature seed for efficient Agrobacterium-mediated soybean transformation. AB - The utility of transformation for soybean improvement requires an efficient system for production of stable transgenic lines. We describe here an improved cotyledonary node method using an alternative explant for Agrobacterium tumefaciens-mediated soybean transformation. We use the term "half-seed" to refer to this alternative cotyledonary explant that is derived from mature seed of soybean following an overnight imbibition and to distinguish it from cotyledonary node derived from 5-7-day-old seedlings. Transformation efficiencies using half seed explants ranged between 1.4 and 8.7% with an overall efficiency of 3.8% based on the number of transformed events that have been confirmed in the T1 generation by phenotypic assay using the herbicide Liberty (active ingredient glufosinate) and by Southern analysis. This efficiency is 1.5-fold higher than the cotyledonary node method used in our laboratory. Significantly, the half-seed system is simple and does not require deliberate wounding of explants, which is a critical and technically demanding step in the cotyledonary node method. PMID- 16249871 TI - High-frequency plant regeneration through cyclic secondary somatic embryogenesis in black pepper (Piper nigrum L.). AB - A high-frequency plantlet regeneration protocol was developed for black pepper (Piper nigrum L.) through cyclic secondary somatic embryogenesis. Secondary embryos formed from the radicular end of the primary somatic embryos which were originally derived from micropylar tissues of germinating seeds on growth regulator-free SH medium in the absence of light. The process of secondary embryogenesis continued in a cyclic manner from the root pole of newly formed embryos resulting in clumps of somatic embryos. Strength of the medium and sucrose concentration influenced the process of secondary embryogenesis and fresh weight of somatic embryo clumps. Full-strength SH medium supplemented with 1.5% sucrose produced significantly higher fresh weight and numbers of secondary somatic embryos while 3.0 and 4.5% sucrose in the medium favored further development of proliferated embryos into plantlets. Ontogeny of secondary embryos was established by histological analysis. Secondary embryogenic potential was influenced by the developmental stage of the explanted somatic embryo and stages up to "torpedo" were more suitable. A single-flask system was standardized for proliferation, maturation, germination and conversion of secondary somatic embryos in suspension cultures. The system of cyclic secondary somatic embryogenesis in black pepper described here represents a permanent source of embryogenic material that can be used for genetic manipulations of this crop species. PMID- 16249870 TI - Codon-modifications and an endoplasmic reticulum-targeting sequence additively enhance expression of an Aspergillus phytase gene in transgenic canola. AB - Transgenic plants offer advantages for biomolecule production because plants can be grown on a large scale and the recombinant macromolecules can be easily harvested and extracted. We introduced an Aspergillus phytase gene into canola (Brassica napus) (line 9412 with low erucic acid and low glucosinolates) by Agrobacterium-mediated transformation. Phytase expression in transgenic plant was enhanced with a synthetic phytase gene according to the Brassica codon usage and an endoplasmic reticulum (ER) retention signal KDEL that confers an ER accumulation of the recombinant phytase. Secretion of the phytase to the extracellular fluid was also established by the use of the tobacco PR-S signal peptide. Phytase accumulation in mature seed accounted for 2.6% of the total soluble proteins. The enzyme can be glycosylated in the seeds of transgenic plants and retain a high stability during storage. These results suggest a commercial feasibility of producing a stable recombinant phytase in canola at a high level for animal feed supplement and for reducing phosphorus eutrophication problems. PMID- 16249872 TI - Rapid and stable buffer exchange system using InSitu Chip suitable for multicolor and large-scale whole-mount analyses. AB - Whole-mount in situ hybridization (WISH) and whole-mount immunohistochemistry (WIHC) are informative methods commonly used to analyze the spatiotemporal and quantitative distribution of mRNAs and proteins. However, these methods require multiple buffer changes and the imposition of time- and nerve-consuming efforts. To facilitate the whole-mount analyses, we innovated an easy and one-step buffer exchange system named "InSitu Chip" based on a single column containing two attached filters. This system improves the speed and stabilizes the different steps of the currently available protocols, providing fast and uniform operations. The InSitu Chip system is especially appropriate for multicolor whole mount analyses using fluorescent detection. Furthermore, the InSitu Chip system is also suitable for large-scale whole-mount experiments associated with genome, transcriptome, and/or proteome analyses requiring high-throughput, high-quality, and reproducible results. Using the InSitu Chip, about 1,500 gene expression patterns were stably surveyed in ascidian Ciona intestinalis juveniles. PMID- 16249873 TI - Subdivision and developmental fate of the head mesoderm in Drosophila melanogaster. AB - In this paper, we define temporal and spatial subdivisions of the embryonic head mesoderm and describe the fate of the main lineages derived from this tissue. During gastrulation, only a fraction of the head mesoderm (primary head mesoderm; PHM) invaginates as the anterior part of the ventral furrow. The PHM can be subdivided into four linearly arranged domains, based on the expression of different combinations of genetic markers (tinman, heartless, snail, serpent, mef 2, zfh-1). The anterior domain (PHMA) produces a variety of cell types, among them the neuroendocrine gland (corpus cardiacum). PHMB, forming much of the "T bar" of the ventral furrow, migrates anteriorly and dorsally and gives rise to the dorsal pharyngeal musculature. PHMC is located behind the T-bar and forms part of the anterior endoderm, besides contributing to hemocytes. The most posterior domain, PHMD, belongs to the anterior gnathal segments and gives rise to a few somatic muscles, but also to hemocytes. The procephalic region flanking the ventral furrow also contributes to head mesoderm (secondary head mesoderm, SHM) that segregates from the surface after the ventral furrow has invaginated, indicating that gastrulation in the procephalon is much more protracted than in the trunk. We distinguish between an early SHM (eSHM) that is located on either side of the anterior endoderm and is the major source of hemocytes, including crystal cells. The eSHM is followed by the late SHM (lSHM), which consists of an anterior and posterior component (lSHMa, lSHMp). The lSHMa, flanking the stomodeum anteriorly and laterally, produces the visceral musculature of the esophagus, as well as a population of tinman-positive cells that we interpret as a rudimentary cephalic aorta ("cephalic vascular rudiment"). The lSHM contributes hemocytes, as well as the nephrocytes forming the subesophageal body, also called garland cells. PMID- 16249874 TI - Towards an in vitro model of cystic fibrosis small airway epithelium: characterisation of the human bronchial epithelial cell line CFBE41o-. AB - The CFBE41o- cell line was generated by transformation of cystic fibrosis (CF) tracheo-bronchial cells with SV40 and has been reported to be homozygous for the DeltaF508 mutation. A systematic characterisation of these cells, which however, is a pre-requisite for their use as an in vitro model, has not been undertaken so far. Here, we report an assessment of optimal culture conditions, the expression pattern of drug-transport-related proteins and the stability/presence of the CF transmembrane conductance regulator (CFTR) mutation in the gene and gene product over multiple passages. The CFBE41o- cell line was also compared with a wild-type airway epithelial cell line, 16HBE14o-, which served as model for bronchial epithelial cells in situ. The CFBE41o- cell line retains at least some aspects of human CF bronchial epithelial cells, such as the ability to form electrically tight cell layers with functional cell-cell contacts, when grown under immersed (but not air-interfaced) culture conditions. The cell line is homozygous for DeltaF508-CFTR over multiple passages in culture and expresses a number of proteins relevant for pulmonary drug absorption (e.g. P-gp, LRP and caveolin-1). Hence, the CFBE41o- cell line should be useful for studies of CF gene transfer or alternative treatment with small drug molecules and for the gathering of further information about the disease at the cellular level, without the need for primary culture. PMID- 16249875 TI - Biodegradation of dichloromethane by the polyvinyl alcohol-immobilized methylotrophic bacterium Ralstonia metallidurans PD11. AB - A dichloromethane (DCM)-degrading bacterium, Ralstonia metallidurans PD11 NBRC 101272, was immobilized in a polyvinyl alcohol (PVA) gel to use in a bioreactor for DCM treatment. After 4-month incubation of PVA gel beads with R. metallidurans PD11 and DCM in a mineral salt medium, the cells were tightly packed in the mesh of the gel. Forty beads of the gel in 10 ml of a batch system model showed effective activity degrading 500 and 1,000 mg l(-1) DCM within 2 and 3 h, respectively. Although reduction of pH due to accumulation of chloride ion liberated from DCM decreased the activity, it was recovered by adjustment to neutral pH. The activity of the immobilized cells was not affected by addition of nutrients which were preferentially utilized by R. metallidurans PD11, unlike the activity of the free-living cells. A continuous flow system with a column was more effective for rapid degradation of DCM. Thus, the PVA gel-immobilized cell of R. metallidurans PD11 is thought to be a prospective candidate to develop the bioreactor. PMID- 16249876 TI - Biotechnological aspects of chitinolytic enzymes: a review. AB - Chitin and chitinases (EC 3.2.1.14) have an immense potential. Chitinolytic enzymes have wide-ranging applications such as preparation of pharmaceutically important chitooligosaccharides and N-acetyl D-glucosamine, preparation of single cell protein, isolation of protoplasts from fungi and yeast, control of pathogenic fungi, treatment of chitinous waste, and control of malaria transmission. In this review, we discuss the occurrence and structure of chitin, the types and sources of chitinases, their mode of action, chitinase production, as well as molecular cloning and protein engineering of chitinases and their biotechnological applications. PMID- 16249877 TI - Growth and denitrifying activity of Xanthobacter autotrophicus CECT 7064 in the presence of selected pesticides. AB - The effects of the application of nine pesticides used commonly in agriculture (aldrin, lindane, dimetoate, methylparathion, methidation, atrazine, simazine, captan and diflubenzuron) on growth, CO2 production, denitrifying activity [as nitrous oxide (N2O) released] and nitrite accumulation in the culture medium by Xanthobacter autotrophicus strain CECT 7064 (Spanish Type Culture Collection) (a micro-organism isolated from a submerged fixed-film) were studied. The herbicide atrazine and the insecticide dimetoate totally inhibited growth and biological activity of X. autotrophicus at 10 mg l(-1), while the rest of the tested pesticides delayed the growth of strain CECT 7064 but did not drastically affect the bacterial growth after 96 h of culture. The denitrifying activity of X. autotrophicus was negatively affected by the pesticides application with the exception of fungicide captan. The release of N2O was strongly inhibited by several pesticides (aldrin, lindane, methylparathion, methidation and diflubenzuron), while dimetoate, atrazine and simazine inhibited totally the denitrifying activity of the strain. The effects of the pesticides on denitrifying submerged fixed-film reactor are discussed. PMID- 16249878 TI - Outlines of an "exploding" network of metabolites generated from the fluoroquinolone enrofloxacin by the brown rot fungus Gloeophyllum striatum. AB - Degradation of the veterinary fluoroquinolone antibiotic enrofloxacin (EFL) was studied with three strains of Gloeophyllum, basidiomycetous fungi thought to produce extracellular hydroxyl radicals. Metabolites generated in a mineral medium were analyzed by combined high-performance liquid chromatography/high resolution electrospray ionization mass spectrometry. Their origin was inferred from peak doublets representing 12C and 14C isotopomers detected at a defined proportion. From each exact molecular mass, the molecular formula was derived for which the most probable chemical structure was postulated, using for guidance 18 known EFL metabolites. All supernatants provided similar metabolite patterns, with the most comprehensive consisting of 87 compounds. These metabolites belonged to five families headed by EFL, its oxidatively decarboxylated or defluorinated congeners, an isatin-, and an anthranilic acid-type derivative. Metabolites hydroxylated in the aromatic part suggested the formation of three catechols and two oxidizable ortho-aminophenol-type compounds. After oxidation to the respective ortho-quinones or ortho-quinone imines and oxidative ring cleavage at one of three alternative sites, the formation of various cis,cis-muconic acid type derivatives is likely, one of which could be detected. Anthranilic acid-type compounds provided two additional sites for ortho-aminophenol formation and aromatic ring cleavage. An "exploding" network of diverse EFL congeners produced by Gloeophyllum suggests the broad utility of our model for studying biodegradation. PMID- 16249879 TI - A rapid and efficient method for multiple-site mutagenesis with a modified overlap extension PCR. AB - A rapid and efficient method to perform site-directed mutagenesis based on an improved version of overlap extension by polymerase chain reaction (OE-PCR) is demonstrated in this paper. For this method, which we name modified (M)OE-PCR, there are five steps: (1) synthesis of individual DNA fragments of interest (with average 20-bp overlap between adjacent fragments) by PCR with high-fidelity pfu DNA polymerase, (2) double-mixing (every two adjacent fragments are mixed to implement OE-PCR without primers), (3) pre-extension (the teams above are mixed to obtain full-length reassembled DNA by OE-PCR without primers), (4) synthesis of the entire DNA of interest by PCR with outermost primers and template DNA from step 3, (5) post-extension (ten cycles of PCR at 72 degrees C for annealing and extension are implemented). The method is rapid, simple and error-free. It provides an efficient choice, especially for multiple-site mutagenesis of DNAs; and it can theoretically be applied to the modification of any DNA fragment. Using the MOE-PCR method, we have successfully obtained a modified sam1 gene with eight rare codons optimized simultaneously. PMID- 16249881 TI - Phototransduction in primate cones and blowfly photoreceptors: different mechanisms, different algorithms, similar response. AB - Phototransduction in primate cones is compared with phototransduction in blowfly photoreceptor cells. Phototransduction in the two cell types utilizes not only different molecular mechanisms, but also different signal processing steps, producing range compression, contrast constancy, and an intensity-dependent integration time. The dominant processing step in the primate cone is a strongly compressive nonlinearity due to cGMP hydrolysis by phosphodiesterase. In the blowfly photoreceptor a considerable part of the range compression is performed by the nonlinear membrane of the cell. Despite these differences, both photoreceptor cell types are similarly effective in compressing the wide range of naturally occurring intensities, and in converting intensity variations into contrast variations. A direct comparison of the responses to a natural time series of intensities, simulated in the cone and measured in the blowfly photoreceptor, shows that the responses are quite similar. PMID- 16249880 TI - Physiology and glomerular projections of olfactory receptor neurons on the antenna of female Heliothis virescens (Lepidoptera: Noctuidae) responsive to behaviorally relevant odors. AB - The neurophysiology and antennal lobe projections of olfactory receptor neurons housed within short trichoid sensilla of female Heliothis virescens F. (Noctuidae: Lepidoptera) were investigated using a combination of cut-sensillum recording and cobalt-lysine staining techniques. Behaviorally relevant odorants, including intra- and inter-sexual pheromonal compounds, plant and floral volatiles were selected for testing sensillar responses. A total of 184 sensilla were categorized into 25 possible sensillar types based on odor responses and sensitivity. Sensilla exhibited both narrow (responding to few odors) and broad (responding to many odors) response spectra. Sixty-six percent of the sensilla identified were stimulated by conspecific odors; in particular, major components of the male H. virescens hairpencil pheromone (hexadecanyl acetate and octadecanyl acetate) and a minor component of the female sex pheromone, (Z)-9 tetradecenal. Following characterization of the responses, olfactory receptor neurons within individual sensilla were stained with cobalt lysine (N=39) and traced to individual glomeruli in the antennal lobe. Olfactory receptor neurons with specific responses to (Z)-9-tetradecenal, a female H. virescens sex pheromone component, projected to the female-specific central large female glomerulus (cLFG) and other glomeruli. Terminal arborizations from sensillar types containing olfactory receptor neurons sensitive to male hairpencil components and plant volatiles were also localized to distinct glomerular locations. This information provides insight into the representation of behaviorally relevant odorants in the female moth olfactory system. PMID- 16249882 TI - Corollary discharge inhibition and audition in the stridulating cricket. AB - The romantic notion of crickets singing on a warm summer's evening is quickly dispelled when one comes ear to ear with a stridulating male. Remarkably, stridulating male crickets are able to hear sounds from the environment despite generating a 100 db song (Heiligenberg 1969; Jones and Dambach 1973). This review summarises recent work examining how they achieve this feat of sensory processing. While the responsiveness of the crickets' peripheral auditory system (tympanic membrane, tympanic nerve, state of the acoustic spiracle) is maintained during sound production, central auditory neurons are inhibited by a feedforward corollary discharge signal precisely timed to coincide with the auditory neurons' maximum response to self-generated sound. In this way, the corollary discharge inhibition prevents desensitisation of the crickets' auditory pathway during sound production. PMID- 16249883 TI - Truncation of the CNS-expressed JNK3 in a patient with a severe developmental epileptic encephalopathy. AB - We have investigated the breakpoints in a male child with pharmacoresistant epileptic encephalopathy and a de novo balanced translocation t(Y;4)(q11.2;q21). By fluorescence in situ hybridisation, we have identified genomic clones from both chromosome 4 and chromosome Y that span the breakpoints. Precise mapping of the chromosome 4 breakpoint indicated that the c-Jun N-terminal kinase 3 (JNK3) gene is disrupted in the patient. This gene is predominantly expressed in the central nervous system, and it plays an established role in both neuronal differentiation and apoptosis. Expression studies in the patient lymphoblastoid cell line show that the truncated JNK3 protein is expressed, i.e. the disrupted transcript is not immediately subject to nonsense-mediated mRNA decay, as is often the case for truncated mRNAs or those harbouring premature termination codons. Over-expression studies with the mutant protein in various cell lines, including neural cells, indicate that both its solubility and cellular localisation differ from that of the wild-type JNK3. It is plausible, therefore, that the presence of the truncated JNK3 disrupts normal JNK3 signal transduction in neuronal cells. JNK3 is one of the downstream effectors of the GTPase regulated MAP kinase cascade, several members of which have been implicated in cognitive function. In addition, two known JNK3-interacting proteins, beta arrestin 2 and JIP3, play established roles in neurite outgrowth and neurological development. These interactions are likely affected by a truncated JNK3 protein, and thereby provide an explanation for the link between alterations in MAP kinase signal transduction and brain disorders. PMID- 16249884 TI - Disruptions of the novel KIAA1202 gene are associated with X-linked mental retardation. AB - The extensive heterogeneity underlying the genetic component of mental retardation (MR) is the main cause for our limited understanding of the aetiology of this highly prevalent condition. Hence we set out to identify genes involved in MR. We investigated the breakpoints of two balanced X;autosome translocations in two unrelated female patients with mild/moderate MR and found that the Xp11.2 breakpoints disrupt the novel human KIAA1202 (hKIAA1202) gene in both cases. We also identified a missense exchange in this gene, segregating with the Stocco dos Santos XLMR syndrome in a large four-generation pedigree but absent in >1,000 control X-chromosomes. Among other phenotypic characteristics, the affected males in this family present with severe MR, delayed or no speech, seizures and hyperactivity. Molecular studies of hKIAA1202 determined its genomic organisation, its expression throughout the brain and the regulation of expression of its mouse homologue during development. Transient expression of the wild-type KIAA1202 protein in HeLa cells showed partial colocalisation with the F actin based cytoskeleton. On the basis of its domain structure, we argue that hKIAA1202 is a new member of the APX/Shroom protein family. Members of this family contain a PDZ and two ASD domains of unknown function and have been shown to localise at the cytoskeleton, and play a role in neurulation, cellular architecture, actin remodelling and ion channel function. Our results suggest that hKIAA1202 may be important in cognitive function and/or development. PMID- 16249886 TI - Dose reduction for CT in children with cystic fibrosis: is it feasible to reduce the number of images per scan? AB - BACKGROUND: Reducing the dose for each CT scan is important for children with cystic fibrosis (CF). OBJECTIVE: To determine whether the number of CT images and therefore the dose per CT scan could be reduced without any significant loss of information in children with CF. MATERIALS AND METHODS: A cohort of children with CF was followed with biennial surveillance CT scans, obtained in inspiration after a voluntary breath-hold as 1-mm thick images at 10-mm intervals from lung apex to base. A random set of 20 baseline CT scans and 10 follow-up CT scans were blinded. Sets of every image (10-mm intervals), every second image (20-mm intervals), every third image (30-mm intervals) and a selection of three and five images were scored randomly using a published CT scoring system by one experienced observer. RESULTS: The 20 subjects were 10 years of age with a range of 3.7-17.6 years at baseline. Fewer CT images resulted in a significantly lower (less abnormal) CT score and the number of patients positive for abnormalities decreased subsequently. At intervals greater than 20 mm no significant change in CT score over 2 years could be detected, while the CT scores at 10-mm (P=0.02) and 20-mm (P=0.02) intervals worsened significantly. CONCLUSIONS: A reduction in the number of inspiratory CT images by increasing the interval between images to greater than 10 mm is not a valid option for radiation dose reduction in children with CF. PMID- 16249887 TI - Intraosseous haemangioma of the ilium. AB - Intraosseous haemangioma, an uncommon benign vascular tumour, is most commonly seen in adults and tends to involve the vertebrae and the skull. Lesions of flat bones are rare and the imaging findings in these patients are non-specific. We report a unique case of intraosseous haemangioma in the ilium of a 7-year-old girl studied by US, radiography, scintigraphy, CT and MRI. PMID- 16249888 TI - Large intradiploic growing skull fracture of the posterior fossa. AB - Growing skull fractures (GSFs) are rare complications of head injury and mostly occur in infancy and early childhood. Location in the posterior fossa and intradiploic development of a GSF is very uncommon. We report a 7-year-old boy with a large, 9 x 7 x 4-cm, occipital intradiploic GSF. The lesion developed progressively over a period of 5 years following a documented occipital linear fracture. This case of a GSF developing from a known occipital linear fracture demonstrates that a GSF may reach a considerable size and, although uncommon, intradiploic development and occipital localization of a GSF is possible. PMID- 16249885 TI - Variation in genes involved in the RANKL/RANK/OPG bone remodeling pathway are associated with bone mineral density at different skeletal sites in men. AB - In order to assess the contribution of polymorphisms in the RANKL (TNFSF11), RANK (TNFRSF11A) and OPG (TNFRSF11B) genes to variations in bone mineral density (BMD), a population-based cohort with 1,120 extreme low hip BMD cases or extreme high hip BMD controls was genotyped on five SNPs. We further explored the associations between these genetic variations and forearm BMDs by genotyping 266 offspring and 309 available parents from 160 nuclear families. A family-based association test was used. Significantly positive associations were found for A163G polymorphisms in the promoter regions of the OPG gene, a missense substitution in exon 7 (Ala192Val) of the RANK gene and rs9594782 SNP in the 5' UTR of the RANKL gene with BMD in men only. Men with TC/CC genotypes of the rs9594782 SNP had a 2.1 times higher risk of extremely low hip BMD (P = 0.004), and lower whole body BMD (P < 0.001). Subjects with the TC genotype of the Ala192Val polymorphism had a 40% reduced risk of having extremely low hip BMD (P < 0.01), and higher whole body BMD (P < 0.01). Subjects with the GG genotype of the A163G polymorphism had a 70% reduced risk of having extremely low hip BMD (P < 0.05), and higher whole body BMD (P < 0.01). Significant gene-gene interactions were also observed among the OPG, RANK and RANKL genes. Our findings suggest that genetic variation in genes involved in the RANKL/RANK/OPG bone remodeling pathway are strongly associated with BMD at different skeletal sites in adult men, but not in women. PMID- 16249889 TI - Knee radiography in the diagnosis of skeletal dysplasias. AB - BACKGROUND: Flattening of the epiphyses of long bones is seen in several skeletal dysplasias and standardized measurements on a radiograph of the knee to detect skeletal dysplasias using this feature have been described. Since then only two other studies in which this method was used have been published, and both included only a small number of children and neither had a control group. In addition, the Dutch National Working Group on Skeletal Dysplasias began to have doubts about the reliability of the method. We therefore decided to re-evaluate its accuracy in a population of children with and without a skeletal dysplasia. OBJECTIVE: To determine the diagnostic value of standardized measurements on conventional AP radiographs of the knee in children with a skeletal dysplasia. SUBJECTS AND METHODS: We measured the distal femoral metaphysis and epiphysis according to the published method on conventional AP radiographs of the knee in 45 healthy children and 52 children with a skeletal dysplasia. We compared graphically the height of the distal femoral epiphysis with its width and with the width of the femoral metaphysis. Receiver operating characteristic (ROC) curves were calculated for each group of children. RESULTS: All graphs showed a considerable overlap between children with a skeletal dysplasia and healthy children. The size of the area under the ROC curves for the different groups was small, varying between 0.567 and 0.653. CONCLUSIONS: This method does not discriminate between children with a skeletal dysplasia and healthy children. We therefore consider it to be of little diagnostic value. PMID- 16249891 TI - A control theory approach to the analysis and synthesis of the experimentally observed motion primitives. AB - Recent experiments on frogs and rats, have led to the hypothesis that sensory motor systems are organized into a finite number of linearly combinable modules; each module generates a motor command that drives the system to a predefined equilibrium. Surprisingly, in spite of the infiniteness of different movements that can be realized, there seems to be only a handful of these modules. The structure can be thought of as a vocabulary of "elementary control actions". Admissible controls, which in principle belong to an infinite dimensional space, are reduced to the linear vector space spanned by these elementary controls. In the present paper we address some theoretical questions that arise naturally once a similar structure is applied to the control of nonlinear kinematic chains. First of all, we show how to choose the modules so that the system does not loose its capability of generating a "complete" set of movements. Secondly, we realize a "complete" vocabulary with a minimal number of elementary control actions. Subsequently, we show how to modify the control scheme so as to compensate for parametric changes in the system to be controlled. Remarkably, we construct a set of modules with the property of being invariant with respect to the parameters that model the growth of an individual. Robustness against uncertainties is also considered showing how to optimally choose the modules equilibria so as to compensate for errors affecting the system. Finally, the motion primitive paradigm is extended to locomotion and a related formalization of internal (proprioceptive) and external (exteroceptive) variables is given. PMID- 16249890 TI - Analytical quantification of the inflammatory cell infiltrate and CD95R expression during treatment of drug-induced toxic epidermal necrolysis. AB - The treatment of drug-induced toxic epidermal necrolysis (TEN) remains unsatisfactory. Intravenous immunoglobulins (IVIg) and intravenous cyclosporin A (CsA) have shown some efficacy in short series of patients. We assessed the effects of IVIg and CsA on TEN lesional and apparently uninvolved skin using standard histology and immunohistochemistry. Cutaneous biopsies were taken from necrotic and clinically uninvolved TEN skin at admission (D1) before any treatment, and after a 5-day treatment (D5). Two IVIg-treated patients (0.75 g/kg/day), two CsA-treated patients (5 mg/kg/day) and two control patients only receiving supportive care were compared. Biopsies were examined by standard histology and immunohistochemistry using antibodies directed to CD68 antigen (macrophages), CD45R0 antigen (activated T lymphocytes), Factor XIIIa (dermal dendrocytes) and the CD95 receptor (apoptosis marker). The different cell densities were evaluated by computerized image analysis. The clinical outcomes with the different treatments were also recorded. There was no obvious difference in the duration of hospitalization in intensive care unit between the three groups but one patient passed away in each of the IVIg- and CsA-groups. At D5, no differences were found between the three groups in the histological and clinical rate of re-epithelialization, and in the evolution of T lymphocyte, macrophage and dendrocyte densities in the epidermis and dermis. However, the expression of the CD95 receptor was similarly and strongly abated at D5 in the epidermis of IVIg- and CsA-treated patients, while it was conversely increased in the two patients under supportive care only. Such a difference was found both in necrotic and uninvolved sites. IVIg and CsA treatments thus appeared to exert no obvious effect on the inflammatory infiltrate, but both abated the expression of the CD95 receptor in the skin of TEN patients. This effect did not seem sufficient to fully reverse the clinical evolution of the disease. It is inferred that IVIg and CsA do not completely abate the TEN process. PMID- 16249892 TI - Can co-activation reduce kinematic variability? A simulation study. AB - Impedance modulation has been suggested as a means to suppress the effects of internal 'noise' on movement kinematics. We investigated this hypothesis in a neuro-musculo-skeletal model. A prerequisite is that the muscle model produces realistic force variability. We found that standard Hill-type models do not predict realistic force variability in response to variability in stimulation. In contrast, a combined motor-unit pool model and a pool of parallel Hill-type motor units did produce realistic force variability as a function of target force, largely independent of how the force was transduced to the tendon. To test the main hypothesis, two versions of the latter model were simulated as an antagonistic muscle pair, controlling the position of a frictionless hinge joint, with a distal segment having realistic inertia relative to the muscle strength. Increasing the impedance through co-activation resulted in less kinematic variability, except for the lowest levels of co-activation. Model behavior in this region was affected by the noise amplitude and the inertial properties of the model. Our simulations support the idea that muscular co-activation is in principle an effective strategy to meet accuracy demands. PMID- 16249893 TI - Cellular localization of estrogen receptor-alpha (ERalpha) and -beta (ERbeta) mRNA in the boar testis. AB - Boar testes synthesize high amounts of estrogens which are known to stimulate several male sexual functions in a variety of extragonadal target tissues. Possible effects within the testis depend on the existence of the estrogen receptor subtypes alpha and beta (ERalpha, ERbeta). The precise cellular localization of these subtypes within the testis was, so far, based mainly on protein expression studies using different antibodies in several species including boars shows contradictory results. Therefore, we investigated the ERalpha and ERbeta gene expression using RT-PCR of testis homogenates and RT-PCR after UV-single cell microdissection combined with in-situ hybridization of four fertile boars with an average age of 32 weeks. Both ERalpha and ERbeta mRNA were found in testis homogenates. Using in-situ hybridization and UV-single cell microdissection ERalpha mRNA was present in type A and type B spermatogonia up to mid-pachytene primary spermatocytes in stage V-VIII and stage I of the seminiferous epithelial cycle, but not in other cells. ERbeta mRNA was found only in Sertoli cells. Interstitial Leydig cells revealed neither ERalpha nor ERbeta mRNA. The data suggest a direct impact of estrogen in the boar on Sertoli cell function via ERbeta and germ cell formation via ERalpha. PMID- 16249894 TI - Tuber size variation and organ preformation constrain growth responses of a spring geophyte. AB - Functional responses to environmental variation do not only depend on the genetic potential of a species to express different trait values, but can also be limited by characteristics, such as the timing of organ (pre-) formation, aboveground longevity or the presence of a storage organ. In this experiment we tested to what degree variation in tuber size and organ preformation constrain the responsiveness to environmental quality and whether responsiveness is modified by the availability of stored resources by exposing the spring geophyte Bunium bulbocastanum to different light and nutrient regimes. Growth and biomass partitioning were affected by initial tuber size and resource availability. On average, tuber weight amounted to 60%, but never less than 30% of the total plant biomass. Initial tuber size, considered an estimate of the total carbon pool available at the onset of treatments, affected plant growth and reproduction throughout the experiment but had little effect on the responsiveness of plants to the treatments. The responsiveness was partly constrained by organ preformation: in the second year variation of leaf number was considerably larger than in the first year of the treatments. The results indicate that a spring geophyte with organ preformation has only limited possibilities to respond to short-term fluctuations of the environment, as all leaves and the inflorescence are preformed in the previous growing season and resources stored in tubers are predominantly used for survival during dormancy and are not invested into plastic adjustments to environmental quality. Such spring geophytes have only limited possibilities to buffer environmental variation. This explains their restriction to habitats characterized by predictable changes of the environmental conditions. PMID- 16249895 TI - Turnover rates of nitrogen stable isotopes in the salt marsh mummichog, Fundulus heteroclitus, following a laboratory diet switch. AB - Nitrogen stable isotopes are frequently used in ecological studies to estimate trophic position and determine movement patterns. Knowledge of tissue-specific turnover and nitrogen discrimination for the study organisms is important for accurate interpretation of isotopic data. We measured delta15 N turnover in liver and muscle tissue in juvenile mummichogs, Fundulus heteroclitus, following a laboratory diet switch. Liver tissue turned over significantly faster than muscle tissue suggesting the potential for a multiple tissue stable isotope approach to study movement and trophic position over different time scales; metabolism contributed significantly to isotopic turnover for both liver and muscle. Nitrogen diet-tissue discrimination was estimated at between 0.0 and 1.2 per thousand for liver and -1.0 and 0.2 per thousand for muscle. This is the first experiment to demonstrate a significant variation in delta15 N turnover between liver and muscle tissues in a fish species. PMID- 16249896 TI - Direct effects of physical stress can be counteracted by indirect benefits: oyster growth on a tidal elevation gradient. AB - The paradigmatic gradient for intertidal marine organisms of increasing physical stress from low to high elevation has long served as the basis for using direct effects of duration of water coverage to predict many biological patterns. Accordingly, changes in potential feeding time may predict the direction and magnitude of differences between elevations in individual growth rates of sessile marine invertebrates. Oysters (triploid Crassostrea ariakensis) experimentally introduced at intertidal (MLW+0.05 m) and subtidal (MLW-0.25 m) elevations in racks provided a test of the ability to use duration of water coverage to predict changes in growth. During early-to-mid winter, a depression of 38-47% in shell growth of intertidal oysters matched the 36% reduction in available feeding time relative to subtidal oysters. In late winter as solar heating of exposed oysters increased, growth differences of 52-55% departed only slightly from the predicted 39%. In spring, however, duration of water coverage failed to predict even the correct direction of growth change with elevation as intertidal oysters grew 34% faster despite 39% less feeding time. Intense seasonal development of shell fouling by other suspension feeders like ascidians, mussels, and barnacles on subtidal (94% incidence) but not on aerially exposed intertidal (21-38% incidence) oysters may explain why duration of water cover failed to predict spring growth differences. Less intense fouling develops on intertidal oysters due to the physiological stress of aerial exposure on settlers, especially during higher temperatures and longer solar exposures of spring. Fouling by suspension feeders is known to reduce growth of the host through localized competition for food and added energetic costs. Thus, in springtime, indirect effects of aerial exposure providing a partial refuge from biological enemies overwhelmed direct effects of reduced duration of water coverage to reverse the expected pattern of slower intertidal growth of a marine invertebrate. PMID- 16249897 TI - Variable responses of old-field perennials to arbuscular mycorrhizal fungi and phosphorus source. AB - If arbuscular mycorrhizal fungi (AMF) promote phosphorus partitioning of plant hosts, they could provide one mechanism for the maintenance of plant community diversity. We investigated whether AMF improved the ability of old field perennials to grow on a range of phosphorus sources and whether AMF facilitated differential performance of plant species on different phosphorus sources (phosphorus niche partitioning). We manipulated form of phosphorus (control versus different inorganic and organic sources) and AM fungal species (control versus four individual AMF species or an AMF community) for five old field perennials grown in a greenhouse in individual culture. Based on biomass after four months of growth, we found no evidence for phosphorus niche partitioning. Rather, we found that effects of AMF varied from parasitic to mutualistic depending on plant species, AMF species, and phosphorus source (significant Plant x Fungus x Phosphorus interaction). Our results suggest that the degree of AMF benefit to a plant host depends not only on AMF species, plant species, and soil phosphorus availability (as has also been found in other work), but can also depend on the form of soil phosphorus. Thus, the position of any AMF species along the mutualism to parasitism continuum may be a complex function of local conditions, and this has implications for understanding plant competitive balance in the field. PMID- 16249898 TI - Testing models of parental investment strategy and offspring size in ants. AB - Parental investment strategies can be fixed or flexible. A fixed strategy predicts making all offspring a single 'optimal' size. Dynamic models predict flexible strategies with more than one optimal size of offspring. Patterns in the distribution of offspring sizes may thus reveal the investment strategy. Static strategies should produce normal distributions. Dynamic strategies should often result in non-normal distributions. Furthermore, variance in morphological traits should be positively correlated with the length of developmental time the traits are exposed to environmental influences. Finally, the type of deviation from normality (i.e., skewed left or right, or platykurtic) should be correlated with the average offspring size. To test the latter prediction, we used simulations to detect significant departures from normality and categorize distribution types. Data from three species of ants strongly support the predicted patterns for dynamic parental investment. Offspring size distributions are often significantly non-normal. Traits fixed earlier in development, such as head width, are less variable than final body weight. The type of distribution observed correlates with mean female dry weight. The overall support for a dynamic parental investment model has implications for life history theory. Predicted conflicts over parental effort, sex investment ratios, and reproductive skew in cooperative breeders follow from assumptions of static parental investment strategies and omnipresent resource limitations. By contrast, with flexible investment strategies such conflicts can be either absent or maladaptive. PMID- 16249899 TI - Effect of continuous lumbar traction on the size of herniated disc material in lumbar disc herniation. AB - We investigated the effects of continuous lumbar traction in patients with lumbar disc herniation on clinical findings, and size of the herniated disc measured by computed tomography (CT). In this prospective, randomized, controlled study, 46 patients with lumbar disc herniation were included, and randomized into two groups as the traction group (24 patients), and the control group (22 patients). The traction group was given a physical therapy program and continuous lumbar traction. The control group was given the same physical therapy program without traction, for the same duration of time. Data for the clinical symptoms and signs were collected before and after the treatment together with calculation of a herniation index, from the CT images that showed the size of the herniated disc material. In the traction group, most of the clinical findings significantly improved with treatment. Size of the herniated disc material in CT decreased significantly only in the traction group. In the traction group the herniation index decreased from 276.6+/-129.6 to 212.5+/-84.3 with treatment (p<0.01). In the control group, pretreatment value was 293.4+/-112.1, and it decreased to 285.4+/-115.4 after the treatment (p>0.05). Patients with greater herniations tended to respond better to traction. In conclusion, lumbar traction is both effective in improving symptoms and clinical findings in patients with lumbar disc herniation and also in decreasing the size of the herniated disc material as measured by CT. PMID- 16249900 TI - In vivo evaluation of early disease progression by X-ray phase-contrast imaging in the adjuvant-induced arthritic rat. AB - PURPOSE: To study the early change of bone matrix and soft tissue around articulation in adjuvant-induced arthritic (AIA) rats non-invasively by X-ray phase-contrast imaging (XPCI), a new imaging method. MATERIALS AND METHODS: Adjuvant-induced arthritis was established in male Sprague-Dawley (SD) rats (n=6, age 40 days) by subcutaneous injection of Freund's complete adjuvant (FCA) into the left hindpaw. In vivo XPCI evaluation of the early soft tissue and bone changes in AIA rats was consecutively performed and correlated with changes in volumes of right hindpaws and body weights. In comparison, the changes in the AIA rats were also evaluated with absorption-contrast imaging using the same X-ray source as XPCI and conventional radiography at the same time. After the imaging evaluation, AIA rats were subjected to histological examination. RESULTS: There was significant difference between the score of XPCI and the other two methods in demonstrating soft tissue (P<0.01), bone details (P<0.01) and lesions (P<0.001). By day 10 after subcutaneous injection of FCA, bone changes in the right hindpaw were not obvious, but swelling of soft tissue appeared. By day 12, bone erosion in the articular facet and the area around the articular facet, was detected, along with osteoporosis, and swelling of soft tissue was aggravated. By day 14 bone erosions became fused and expanded, especially in the margin area around the articular facet. At day 16 bone erosion still existed. Joint interspaces seemed wider than normal, and swelling of soft tissue was significant. By day 18 periosteal new bone formation was seen definitely, destruction of bone decreased, bone density around the articular was enhanced, and swelling of soft tissue was relieved. XPCI could clearly distinguish all these alterations, which could not be demonstrated by absorption-contrast imaging and conventional radiography. During the test period, the volume of the right hindpaw and the body weight of the AIA rats also changed significantly compared with the normal rat. Histological examination confirmed that adjuvant-induced arthritis had occurred in all rats of the adjuvant group. CONCLUSION: Osteoporosis, bone erosion and periosteal new bone formation take place at the early stage of adjuvant-induced arthritis. XPCI can evaluate non-invasively these subtle bone changes that are "blind areas" for conventional radiography. PMID- 16249901 TI - Surgical treatment for injuries of the middle and lower cervical spine. AB - We reviewed the surgical treatment of 31 patients with burst fractures or teardrop dislocation fractures in the middle and lower cervical spine. Patients were treated with anterior instrumentation, posterior instrumentation, or a combination of both. Patients were evaluated radiographically and with the Frankel neurological outcomes grading scale. Anterior decompression and fusion restored the spinal canal diameter by approximately 60% whereas the posterior or combined approaches restored the canal diameter by only 6%. In addition, nine of 24 patients treated anteriorly gained improved neurological function whereas none of the patients treated posteriorly had neurological improvement. Based on the anatomical and neurological findings, the study demonstrates that anterior fusion is preferable to posterior fusion for the treatment of burst fractures and tear drop dislocation fractures of the middle and lower cervical spine. PMID- 16249902 TI - Further experience with OK-432 for lymphangiomas. AB - This study includes all the children treated with OK-432 for lymphangioma at our institute. Twenty-nine children treated between 1999 and 2003 are reported for the first time: twelve cases regressed completely, eight cases regressed more than 50% and seven remained unchanged; two cases were lost at follow-up. The outcome was related to the size of the cysts, the larger ones having a better prognosis. The adverse reactions are discussed and the methods of treatment are described in detail. Fifteen children, treated before 1999 and already reported, are reviewed after a long-term follow-up. Four had a recurrence: one regressed spontaneously and three needed further treatment. The other 11 had no complaints. Even considering the risk of recurrence, OK-432 therapy remains our first line therapy for lymphangiomas, avoiding surgery in most cases. PMID- 16249903 TI - Treatment of chondral defects by hydro jet. Results of a preliminary scanning electron microscopic evaluation. AB - INTRODUCTION: Arthroscopic methods in treatment of chondral defects aim to get smooth cartilage surfaces. Mechanical and thermal methods are used. Often the clinical results are poor or moderate. The treatment of chondral defects by using a hydro jet is an innovative measure. This study was aimed to evaluate the quality of the chondral surfaces after mechanical, thermal and hydro jet treatments in an in vitro scanning electron microscopy (SEM) study. MATERIALS AND METHODS: Femoral osteochondral pieces of the lateral condyle were obtained intraoperatively from patients undergoing total knee arthroplasty for primary knee OA. Partial-thickness cartilage degree II defects were smoothed by a mechanical shaver, bipolar radio frequency energy (RF) and hydro jet (84.1 KPa). SEM was carried out to evaluate the effects of the treatment. RESULTS: Mechanical shaving produces a rough surface with groves and open lying collagen fibers. The surface, after bipolar cool ablation (coablation) was also uneven. The matrix was destroyed by massive vacuolization. Hydro jet treatment produces a relatively smooth surface without open lying collagen fibers. CONCLUSION: It is not possible to produce even surfaces by mechanical shaving or bipolar treatment. Hydro jet treatment allows a precise cutting which causes a relatively smooth chondral surface. PMID- 16249904 TI - Unaware of a giant serous cyst adenoma: a case report. AB - A case of 36-year-old nonmarried virgin woman presenting a giant ovarian serous cyst adenoma weighing 9.5 kg is reported here. Ovarian neoplasms may be divided by origin cell type into three main groups: epithelial, stromal and germ cell. Taken as a group, the epithelial tumors are by far the most common type. The single most common benign ovarian neoplasm is the benign cystic teratoma; however, according to some studies it is serous cyst adenoma. A 36-year-old nonmarried virgin woman was referred to our clinic from a local medical center. When she was seen first at our outpatient clinic, she looked like a 9-month pregnant woman. Her medical history was normal. She had no serious illness or operation before. On abdominal ultrasound, a giant cyst was found which encompassed the whole abdomen. At laparotomy, a giant, totally cystic, vascularized and smooth mass attached to the right ovary was encountered, lying between the symphysis and the xiphoid. Ooferectomy was performed. On the postoperative second day, she was discharged without any problem. Her pathology report disclosed a 35 x 20 x 16 cm(3) serous cyst adenoma weighing 9.5 kg. This is the largest ovarian cyst that ever reported from our hospital and one of the largest among the reported cases in the literature. PMID- 16249905 TI - Impact of laparotomy and liver resection on the peritoneal concentrations of fibroblast growth factor 2, vascular endothelial growth factor and hepatocyte growth factor. AB - PURPOSE: Some data have suggested that major surgery is associated with the post operative growth of residual tumour masses but the mechanism of this is unknown. This study was designed to determine the relationship between intraperitoneal (IP) cytokine levels, and laparotomy in benign and malignant settings. METHODS: Intraperitoneal fluid specimens were obtained at the start and at the end of laparotomy in patients with benign conditions (n=10) and in others undergoing resection of hepatic metastases from colorectal cancer (n=10). Using ELISA the concentration of the angiogenic cytokines, HGF, VEGF-A, VEGF-C, VEGF-D and FGF-2 was determined. RESULTS: The data show that in 16 of 20 patients there was a significant increase (P=0.006) in the IP concentration of hepatocyte growth factor (HGF) but not in the other growth factors by the end of the operation. The mean increase in HGF concentration was 821.5 pg/ml (95% CI: 11.0-6,426.0). Neither the groups (malignant and non-malignant) nor the length of operation correlated with greater or lesser increases in HGF. CONCLUSION: The observation that the increase in HGF occurred in both the cancer and non-cancer groups suggests that it is the surgery rather than the disease that is associated with the increased cytokine concentration. As HGF is a potent endothelial, epithelial and mesenchymal mitogen the data highlight HGF as a potential target for anti cancer treatments in the peri-operative period. However, investigators should closely monitor wound healing as this may be compromised by this new class of drugs. PMID- 16249906 TI - Relevance of endogenous 3alpha-reduced neurosteroids to depression and antidepressant action. AB - The naturally occurring 3alpha-reduced neurosteroids allopregnanolone and its isomer pregnanolone are among the most potent positive allosteric modulators of gamma-aminobutyric acid type A receptors. They play a critical role in the maintenance of physiological GABAergic tone and display a broad spectrum of neuropsychopharmacological properties. We have reviewed existing evidence implicating the relevance of endogenous 3alpha-reduced neuroactive steroids to depression and to the mechanism of action of antidepressants. A wide range of preclinical and clinical evidence suggesting the antidepressant potential of 3alpha-reduced neuroactive steroids and a possible involvement of a deficiency and a disequilibrium of neuroactive steroid levels in pathomechanisms underlying the etiology of major depressive disorder have emerged in recent years. Antidepressants elevate 3alpha-reduced neurosteroid levels in rodent brain, and clinically effective antidepressant pharmacotherapy is associated with normalization of plasma and cerebrospinal fluid (CSF) concentrations of endogenous neuroactive steroids in depressed patients, unveiling a possible contribution of neuroactive steroids to the mechanism of action of antidepressants. In contrast, recent studies using nonpharmacological antidepressant therapy suggest that changes in plasma neuroactive steroid levels may not be a general mandatory component of clinically effective antidepressant treatment per se, but may reflect distinct properties of pharmacotherapy only. While preclinical studies offer convincing evidence in support of an antidepressant-like effect of 3alpha-reduced neuroactive steroids in rodent models of depression, current clinical investigations are inconclusive of an involvement of neuroactive steroid deficiency in the pathophysiology of depression. Moreover, clinical evidence is merely suggestive of a role of neuroactive steroids in the mechanism of action of clinically effective antidepressant therapy. Additional clinical studies evaluating the impact of successful pharmacological and nonpharmacological antidepressant therapies on changes in neuroactive steroid levels in both plasma and CSF samples of the same patients are necessary in order to more accurately address the relevance of 3alpha-reduced neuroactive steroids to major depressive disorder. Finally, proof of-concept studies with drugs that are known to selectively elevate brain neurosteroid levels may offer a direct assessment of an involvement of neurosteroids in the treatment of depressive symptomatology. PMID- 16249908 TI - Dissociating the primary reinforcing and reinforcement-enhancing effects of nicotine using a rat self-administration paradigm with concurrently available drug and environmental reinforcers. AB - RATIONALE: Nicotine has two effects on reinforcement in traditional self administration paradigms. It serves as a primary reinforcer by increasing the probability of behaviors that result in nicotine delivery. However, nicotine also potently enhances behaviors that result in the delivery of nonpharmacological reinforcers. OBJECTIVES: The present study sought to dissociate these two effects of nicotine on reinforcement. METHODS: For one group of rats (2 lever), a nonpharmacological reinforcer [visual stimulus (VS)] was available for pressing one lever. Nicotine infusions were available for pressing a different lever. A second group (NIC + VS) received more traditional self-administration training; both the VS and nicotine were delivered for pressing a single active lever. Control groups received either nicotine infusions (NIC only) or VS presentations (VS only) for pressing the active lever. RESULTS: Nicotine alone was a weak reinforcer; the VS alone was slightly more reinforcing than nicotine. When these two reinforcers were combined (NIC + VS), response rates were synergistically increased. For the 2-lever group, responding on the nicotine lever was weak, matching the response rates of rats receiving nicotine alone. However, responding on the VS lever was potently enhanced in this group; equaling the response rates for rats receiving both reinforcers for making a single response (NIC + VS). CONCLUSIONS: These data indicate that the reinforcement-enhancing effects of nicotine are very potent even when only moderate quantities of the drug are self administered. Moreover, they provide the first demonstration that the reinforcement-enhancing and primary reinforcing effects of nicotine can be dissociated behaviorally. PMID- 16249907 TI - Intense cocaine self-administration after episodic social defeat stress, but not after aggressive behavior: dissociation from corticosterone activation. AB - RATIONALE: An intense stress response characterizes both the dominant and submissive individuals during an aggressive confrontation, and these stress responses have enduring neural and behavioral consequences. OBJECTIVES: In spite of similar glucocorticoid and corticolimbic dopamine activation, dominant and defeated individuals appear to diverge in terms of their drug taking. Do rats that are intermittently subjected to defeat stress become more sensitized to cocaine taking relative to rats that engage in aggressive bouts? METHODS: Separate groups of male Long-Evans rats were investigated after an initial 10-day period with four brief episodes of social defeat (intruders) or aggressive behavior (residents): (1) the corticosterone responses to the very first and the last confrontations were measured; (2) the locomotor response to an amphetamine (1 mg/kg) challenge 10 days after the last stress exposure served as an index of behavioral sensitization; (3) intravenous self-administration sessions assessed the reinforcing effects of 0.75 mg/kg/infusion cocaine when available after every fifth response (fixed ratio), when delivered after completing progressively more demanding response requirements (progressive ratio; 0.3 mg/kg/infusion), and when available during a 24-h binge of continuous access (0.3 mg/kg/infusion). RESULTS: Both social defeat of the intruder rat and attack behavior by the resident rat rapidly increased plasma levels of corticosterone after the first and last aggressive confrontation, indicating no habituation to these types of stress. Intermittent social defeat engenders a sensitized locomotor response to a 1 mg/kg amphetamine challenge and increases cocaine self-administration as indicated by more behavioral effort to obtain cocaine infusions and by accumulating more cocaine during 24 h of continuous access (binge). By contrast, experiences with aggressive behavior do not impact on the motorically activating and reinforcing effects of stimulant administrations. CONCLUSIONS: The closely similar corticosterone activation in dominant and subordinate rats, followed by divergent patterns of cocaine self-administration indicates that different forms of social stress have dissociable effects on cocaine taking. PMID- 16249909 TI - Preliminary evidence for medication effects on functional abnormalities in the amygdala and anterior cingulate in bipolar disorder. AB - RATIONALE: Abnormal amygdala and frontocortical responses to emotional stimuli are implicated in bipolar disorder (BD) and have been proposed as potential treatment targets. OBJECTIVES: The aim of this study was to investigate amygdala and frontocortical responses to emotional face stimuli in BD and the influences of mood-stabilizing medications on these responses. METHODS: Functional magnetic resonance imaging was performed while 17 BD participants (5 unmedicated) and 17 healthy comparison (HC) participants viewed faces with happy, sad, fearful, or neutral expressions. RESULTS: The group by stimulus-condition interaction was significant (p<0.01) for amygdala activation, with the greatest effects in the happy face condition. Relative to HC, amygdala increases were greater in unmedicated BD, but lower in medicated BD. Rostral anterior cingulate (rAC) activation was decreased in unmedicated BD compared to HC; however, BD participants taking medication demonstrated rAC activation similar to HC participants. CONCLUSIONS: Although the sample sizes were small, these preliminary results suggest that BD is associated with increased amygdala and decreased rAC response to emotional faces. The findings also provide preliminary evidence that mood-stabilizing medications may reverse abnormalities in BD in the response of an amygdala-frontal neural system to emotional stimuli. PMID- 16249910 TI - The association of Blastocystis hominis and Endolimax nana with diarrheal stools in Zambian school-age children. AB - To determine the prevalence of endoparasites and their association with diarrhea, a survey was conducted in the Southern Province of Zambia that used conventional and molecular techniques applied to stool and urine samples from school-age children (n = 93). Almost half of the stools (49.5%) were diarrhetic. The overall prevalence of Endolimax nana, Schistosoma haematobium, Blastocystis hominis, Giardia lamblia, Cryptosporidium parvum, Encephalitozoon intestinalis, and Strongyloides stercoralis was 64.3, 59.1, 53.8, 19.4, 8.6, 8.6, and 1.1%, respectively. Only the associations between infection with B. hominis and E. nana with diarrhea were statistically significant. Although B. hominis and E. nana are considered to be nonpathogenic organisms, this study demonstrated that they can be associated with diarrhea in children when they occur at high prevalence and intensity. This survey supports the recent evidence that B. hominis and E. nana infections are associated with deficient sanitation and low hygiene standards and can contribute to diarrhea in children in developing countries. PMID- 16249911 TI - Unravelling the evolution of the head lice and body lice of humans. AB - Recent studies of mitochondrial genes of the head and body lice of humans indicate that present-day lice comprise two lineages that diverged before the evolution of modern humans. To test if this was a locus-specific phenomenon, we studied two nuclear genes, elongation factor-1alpha (EF-1alpha) and small subunit ribosomal RNA (ssu rRNA). Our ssu rRNA phylogeny was concordant with the phylogenies from mitochondrial genes, but the EF-1alpha phylogeny was not concordant either with the mitochondrial phylogenies or with the ssu rRNA phylogeny. So both nuclear (ssu rRNA) and mitochondrial data indicate that there are two lineages of lice: one lineage with head lice only (H-only lineage) the other lineage with head and body lice (H+B lineage). Thus, body lice apparently evolved from just one of the two main lineages of lice. However, the date of divergence and geographical origins of the two lineages are controversial. Kittler et al. (Curr Biol 13:1414-1417, 2003; Curr Biol 14:2309, 2004) proposed that these two lineages diverged 0.77 mya, whereas Reed et al. (PLoS Biol 2:e340, 2004) proposed that they diverged 1.18 mya and suggested that one of the lineages, the H-only lineage, evolved in the New World on Homo erectus. We discuss this hypothesis in light of our results from ssu rRNA. PMID- 16249912 TI - Evaluation of robotic training forces that either enhance or reduce error in chronic hemiparetic stroke survivors. AB - This investigation is one in a series of studies that address the possibility of stroke rehabilitation using robotic devices to facilitate "adaptive training." Healthy subjects, after training in the presence of systematically applied forces, typically exhibit a predictable "after-effect." A critical question is whether this adaptive characteristic is preserved following stroke so that it might be exploited for restoring function. Another important question is whether subjects benefit more from training forces that enhance their errors than from forces that reduce their errors. We exposed hemiparetic stroke survivors and healthy age-matched controls to a pattern of disturbing forces that have been found by previous studies to induce a dramatic adaptation in healthy individuals. Eighteen stroke survivors made 834 movements in the presence of a robot-generated force field that pushed their hands proportional to its speed and perpendicular to its direction of motion--either clockwise or counterclockwise. We found that subjects could adapt, as evidenced by significant after-effects. After-effects were not correlated with the clinical scores that we used for measuring motor impairment. Further examination revealed that significant improvements occurred only when the training forces magnified the original errors, and not when the training forces reduced the errors or were zero. Within this constrained experimental task we found that error-enhancing therapy (as opposed to guiding the limb closer to the correct path) to be more effective than therapy that assisted the subject. PMID- 16249913 TI - CYP2D6 genotype and phenotype determination in a Mexican Mestizo population. AB - OBJECTIVE: Although CYP2D6 genetic polymorphism plays an important role in interindividual and interethnic variability in drug response, very few pharmacogenetic data are available from Hispanic populations, including Mexicans. For this purpose, this study was undertaken to determine CYP2D6 genotype and phenotype in a healthy Mexican Mestizo population. METHODS: Genotyping of five CYP2D6 mutant alleles by PCR-RFLP, and CYP2D6*5 and duplicated CYP2D6 alleles by long-PCR was performed in two hundred and forty three Mexican Mestizos. Of these, one hundred subjects were also phenotyped using dextromethorphan as the probe drug. RESULTS: The frequency of CYP2D6*2, *3, *4, *5, *10, *17 was 19.34%, 1.44%, 11.21%, 2.67%, 12.45%, and 1.65%, respectively, while duplicated CYP2D6 alleles were found in 12.76% of the 243 genotyped subjects. Among the 100 phenotyped subjects, we identified ten (10%, 95% confidence interval of 4.12-15.9) individuals as poor metabolizers by using the published antimode for Caucasians. The mean log10 dextromethorphan/dextrorphan ratio of the total sample was -2.05. The mean (SD) of the log10 MR in the CYP2D6 subgroups was UM = -2.6 (0.86); EM = 2.09 (0.98); IM = -1.71 (1.06); and PM = 0.42 (0.625). These data show a trend toward a smaller mean log MR (higher enzyme activity) as the number of active alleles increases. CONCLUSIONS: The PM frequency of CYP2D6 in the population studied was 10%, which is very similar to Spanish Caucasians. The observed frequency of the CYP2D6 alleles tested was unique for the Mexican Mestizo sample analyzed, and in accordance to the Caucasian, Asian and African admixture in this population. PMID- 16249914 TI - Different pharmacokinetics of tramadol in mothers treated for labour pain and in their neonates. Towards an increased knowledge of paediatric clinical pharmacology. PMID- 16249916 TI - Prediction of charge-induced molecular alignment: residual dipolar couplings at pH 3 and alignment in surfactant liquid crystalline phases. AB - Recently we reported that the alignment tensor of a biological macromolecule, which was dissolved in a dilute suspension of highly negatively charged filamentous phage at close to neutral pH, can be predicted from the molecule's 3D charge distribution and shape (Zweckstetter et al. 2004). Here it is demonstrated that this approach is also applicable to alignment of proteins in liquid crystalline phases formed by filamentous phage at low pH. Residual dipolar couplings (RDCs) predicted by our simple electrostatic model for the B1 domain of protein G in fd phage at pH 3 fit very well with the experimental values. The sign of charge-shape predicted one-bond (1)H-(15)N dipolar couplings for the B1 domain of protein G (GB1) was inverted at pH 3 compared to neutral pH, in agreement with experimental observations. Our predictions indicate that this is a feature specific for GB1. In addition, it is shown that RDCs induced in the protein ubiquitin by the presence of a positively charged surfactant system comprising cetylpyridinium bromide/hexanol/sodium bromide can be predicted accurately by a simple electrostatic alignment model. This shows that steric and electrostatic interactions dominate weak alignment of biomolecules for a wide range of pH values both in filamentous phage and in surfactant liquid crystalline phases. PMID- 16249917 TI - Effects of menstrual phase on performance and recovery in intense intermittent activity. AB - Game sport and training require repeated high intensity bursts. This study examined differences between high intensity, intermittent work in two phases of the menstrual cycle. Six physically active young women (age 19-29) performed 10 6 s sprints on a cycle ergometer in both the mid-follicular (FP) (days 6-10) and late-luteal phases (LP) (days 20-24) of the menstrual cycle. Work, power, oxygen intake (VO2) parameters, and capillarized blood lactate were measured. Data are analyzed using the Friedman and Wilcoxon matched pairs tests. There was no difference between menstrual phases in peak 6-s power (6.8(0.6) W kg(-1) in FP, 6.9(0.6) W kg(-1) in LP), the drop off in work (1.2(3.5) J kg(-1) in FP and 1.0(2.7) J kg(-1) in LP), or in the sprint VO2 (23.7(1.5) mL kg(-1) min(-1) in LP and 24.3(2.4) mL kg(-1) min(-1) in FP). Capillarized blood lactate was also similar in both phases of the menstrual cycle both at 1 min (9.2(2.7) mmol L(-1) in FP, 9.2(3.1) mmol L(-1)) and at 3 min (9.0(2.2) mmol L(-1) in FP, 9.2(2.2) mmol L(-1) in LP). However, the average 6-s work was greater in the LP (39.3(3.4) J kg(-1)) than during the FP (38.3(3.1) J kg(-1)) (P=0.023). The recovery VO2 was also greater in the LP than the FP (26.3(2.4) mL kg(-1) min(-1) in LP, 25.0(2.6) mL kg(-1) min(-1) in FP, P=0.023). Average work over a series of sprints and the VO2 consumed between sprints may be slightly greater during the LP than the FP of the menstrual cycle. PMID- 16249918 TI - Influence of maximal muscle strength and intrinsic muscle contractile properties on contractile rate of force development. AB - 'Explosive' muscle strength or contractile rate of force development (RFD) is a term to describe the ability to rapidly develop muscular force, and can be measured as the slope of the torque-time curve obtained during isometric conditions. Previously, conflicting results have been reported regarding the relationship between contractile RFD and various physiological parameters. One reason for this discrepancy may be that RFD in various time intervals from the onset of contraction is affected by different physiological parameters. The aim of the present study was to investigate the relationship between voluntary contractile RFD in time intervals of 0-10, 0-20, ..., 0-250 ms from the onset of contraction and two main parameters: (1) voluntary maximal muscle strength and (2) electrically evoked muscle twitch contractile properties. The main finding was that voluntary RFD became increasingly more dependent on MVC and less dependent on muscle twitch contractile properties as time from the onset of contraction increased. At time intervals later than 90 ms from the onset of contraction maximal muscle strength could account for 52-81% of the variance in voluntary RFD. In the very early time interval (<40 ms from the onset of contraction) voluntary RFD was moderately correlated to the twitch contractile properties of the muscle and was to a less extent related to MVC. The present results suggest that explosive movements with different time spans are influenced by different physiological parameters. This may have important practical implications when designing resistance training programs for specific sports. PMID- 16249919 TI - Changes in fluctuation of isometric force following eccentric and concentric exercise of the elbow flexors. AB - This study tested the hypothesis that eccentric exercise (ECC) would increase force fluctuation for several days following exercise; however, concentric exercise (CON) would not produce such an effect. Twelve men performed six sets of five reps of dumbbell exercise of the elbow flexors eccentrically with one arm and concentrically with the other, separated by 4-6 weeks, using a dumbbell set at 50% of maximal voluntary isometric contraction (MVC) measured at 90 degrees of elbow flexion. MVC, range of motion (ROM), upper arm circumference, plasma creatine kinase activity (CK), myoglobin concentration (Mb) and muscle soreness were assessed before, immediately after, 1 h and 1-5 days following both exercise bouts. Force fluctuations during 30, 50 and 80% MVC were quantified by coefficient of variation (CV) of the force data (sampling frequency: 100 Hz) for 4 s. Significantly (P < 0.01) larger changes in MVC, ROM, and upper arm circumference were evident following ECC compared to CON, and only ECC resulted in significant (P < 0.01) increases in CK and Mb, and development of muscle soreness. Significant (P < 0.01) differences existed between ECC and CON for changes in force fluctuations. CV increased significantly (P < 0.01) immediately and 1 h after ECC from baseline for 30, 50, and 80% MVC without a significant difference among the intensities, and no significant changes in CV were evident following CON. It was concluded that increases in force fluctuation were peculiar to ECC, but did not necessarily reflect muscle damage. PMID- 16249920 TI - Alternative strategies for exercise critical power estimation in patients with COPD. AB - Exercise critical power (CP) has been shown to represent the highest sustainable work rate (WR) in patients with chronic obstructive pulmonary disease (COPD). Parameter estimation, however, depends on 4 high-intensity tests performed, on different days, to the limit of tolerance (T(lim)). In order to establish a milder protocol that would be more suitable for disabled patients, we contrasted CP derived from 4, 3 and 2 tests (CP4, CP3 and CP2) in 8 males with moderate COPD. In addition, CP was calculated from 2 single-day tests performed on an inverse sequence (CP(2AB) and CP(2BA)): CP values within 5 W from CP4 were assumed as "clinically-acceptable" estimates. We found that [CP4-CP3] and [CP4 CP2] differences were within 5 W in 8 and 6 patients, respectively (95% confidence interval of the differences = -1.3 to 3.5 W and -11.5 to 6.5 W). There was a systematic decline on T(lim) when an exercise bout was performed after a previous test on the same day (P<0.05). Consequently, substantial differences were found between CP4 and any of the CP estimates obtained from single-day tests. In conclusion, clinically-acceptable estimates of CP can be obtained by using 3 or, in most circumstances, 2 constant WR tests in patients with moderate COPD--provided that they are not performed on the same day. PMID- 16249921 TI - Iron-regulatory protein hepcidin is increased in female athletes after a marathon. AB - The propose of this study was to determine the influence of marathon race on hepcidin excretion in female athletes (age 26-45 years). Urine samples were taken before, immediately after, 1 and 3 days after the race. In the average, hepcidin transiently increased at day 1 from 32 to 85 ng/mg creatinine. We propose that the frequently observed iron deficiency of females runners is caused by elevated hepcidin levels. PMID- 16249922 TI - Climate controls on valley fever incidence in Kern County, California. AB - Coccidiodomycosis (valley fever) is a systemic infection caused by inhalation of airborne spores from Coccidioides immitis, a soil-dwelling fungus found in the southwestern United States, parts of Mexico, and Central and South America. Dust storms help disperse C. immitis so risk factors for valley fever include conditions favorable for fungal growth (moist, warm soil) and for aeolian soil erosion (dry soil and strong winds). Here, we analyze and inter-compare the seasonal and inter-annual behavior of valley fever incidence and climate risk factors for the period 1980-2002 in Kern County, California, the US county with highest reported incidence. We find weak but statistically significant links between disease incidence and antecedent climate conditions. Precipitation anomalies 8 and 20 months antecedent explain only up to 4% of monthly variability in subsequent valley fever incidence during the 23 year period tested. This is consistent with previous studies suggesting that C. immitis tolerates hot, dry periods better than competing soil organisms and, as a result, thrives during wet periods following droughts. Furthermore, the relatively small correlation with climate suggests that the causes of valley fever in Kern County could be largely anthropogenic. Seasonal climate predictors of valley fever in Kern County are similar to, but much weaker than, those in Arizona, where previous studies find precipitation explains up to 75% of incidence. Causes for this discrepancy are not yet understood. Higher resolution temporal and spatial monitoring of soil conditions could improve our understanding of climatic antecedents of severe epidemics. PMID- 16249923 TI - [NATO mission in Kosovo: historical backgrounds and informations of working as radiologist in the German field hospital]. AB - The first part of this article describes how the NATO mission in Kosovo came into existence and focuses on the historical background and ethnical problems. The second part deals with the working conditions of a radiologist in the German field hospital in Prizren and focuses on the personnel and technical equipment in the radiological department. PMID- 16249924 TI - [Case report: massive lower intestinal bleeding from ileal varices: treatment with transjugular intrahepatic portosystemic shunt (TIPSS)]. AB - Acute gastrointestinal bleeding in patients with liver cirrhosis is associated with a high mortality. Ileal varices and collaterals from ectopic vessels are extremely rare, encountered in less than 5% of the cirrhotic patients. The diagnosis is frequently delayed because the regular diagnostic methods such as gastroscopy or colonoscopy are unsuccessful in accurate the source of bleeding in the majority of the cases. We report an unusual case of massive and uncontrollable lower intestinal bleeding from ileal varices with right ovarian vein anastomosis in a 56 year-old female patient with liver cirrhosis and previous history of abdominal and pelvic surgery. The accurate angiographic and computed tomography diagnosis allowed fast decompression of the portal venous system using a transjugular intrahepatic portosystemic shunt. PMID- 16249925 TI - The effect of N-acetylcysteine on posttraumatic changes after controlled cortical impact in rats. AB - OBJECTIVE: The antioxidant potential N-Acetylcysteine (NAC) and its improvement of posttraumatic mitrochondrial dysfunction have been reported. This study investigated the effect of NAC on posttraumatic changes after controlled cortical Impact (CCI) injury. DESIGN AND SETTING: Prospective randomized controlled animal study. METHODS: A moderate left focal cortical contusion was induced using CCI. Either NAC (163 mg/kg bw) or physiological saline was administered intraperitoneally immediately and 2 and 4 h after trauma. Blood gases, temperature, mean arterial blood pressure (MABP), and intracranial pressure (ICP) were monitored. Twenty-four hours after trauma brains were removed and either posttraumatic edema was quantified gravimetrically (n=24], or contusion volume was determined morphometrically using slices staining and computerized image analysis (n=24]. Laser Doppler flowmetry was used to assess pericontusional cortical perfusion before trauma, 30 min and 4 and 24 h after trauma (n=14]. MEASUREMENTS AND RESULTS: Physiological parameters remained within normal limits. ICP measurements and water content in traumatized hemispheres did not differ between the groups. Relative contusion volume of the left hemisphere was slightly but nonsignificantly diminished in NAC-treated animals (4.7+/-0.4% vs. 5.9+/-0.5% in controls). In both groups pericontusional perfusion was significantly reduced at 4 h followed by a state of hyperperfusion at 24 h with no differences between the groups. CONCLUSIONS: Despite previously reported neuroprotective abilities of NAC, no positive effect on posttraumatic perfusion, brain edema formation, or contusion volume after focal brain injury was observed in this study. PMID- 16249926 TI - Static pressure volume curves and body posture in acute respiratory failure. AB - OBJECTIVE: In acute respiratory distress syndrome the body posture effects on pressure-volume (PV) curves are still unclear. We examined the effects of prone position on inflation PV curves and their potential relationships with postural alterations in gas exchange. DESIGN AND SETTING: Prospective study with patients serving as their own controls in a university-affiliated 30-bed intensive care unit. PATIENTS AND PARTICIPANTS: Thirteen anesthetized, paralyzed, semirecumbent, mechanically ventilated patients with early/severe/diffuse ARDS. INTERVENTIONS: Sequential body posture changes: preprone semirecumbent, prone, and postprone semirecumbent. MEASUREMENTS AND RESULTS: In each posture hemodynamics, gas exchange, and lung volumes were determined before/during removal and after restoration of positive end-expiratory pressure (PEEP=10.2+/-0.6 cmH2O). At zero PEEP PV curves of respiratory system, lung, and chest wall were constructed. Prone position vs. preprone semirecumbent resulted in significantly reduced pressure at lower inflection point of lung PV curve (2.2+/-0.2 vs. 3.7+/-0.5 cmH2O) and increased volume at upper inflection point (0.87+/-0.03 vs. 0.69+/ 0.05 l). Postural reduction in lower inflection point pressure of lung PV curve was the sole independent predictor of pronation-induced increases in PaO2/FIO2 (R2=0.76). PaO2/FIO2 increases were also significantly related with increases in functional residual capacity (R2=0.60). CONCLUSIONS: In early/severe/diffuse ARDS prone position reduces lower inflection point pressure and increases upper inflection point UIP volume of the lung PV curve. Lower inflection point pressure reductions explain oxygenation improvements, which are also associated with a postural increase in functional residual capacity. PMID- 16249927 TI - A nationwide survey of intensive care unit discharge practices. AB - OBJECTIVE: To describe intensive care unit (ICU) discharge practices, examine factors associated with physicians' discharge decisions, and explore ICU and hospital characteristics and clinical determinants associated with the discharge process. DESIGN: Survey in adult ICUs affiliated with the Swiss Society of Intensive Care Medicine. INTERVENTIONS: Questionnaire inquiring about ICU structure and organization mailed to 73 medical directors. Level of monitoring, intravenous medications, and physiological variables were proposed as elements of discharge decision. Five clinical situations were presented with request to assign a discharge disposition. MEASUREMENTS AND RESULTS: Fifty-five ICUs participated, representing 75% of adult Swiss ICUs. Responsibility for patient management was assigned in 91% to the ICU team directing patient care. Only 22% of responding centers used written discharge guidelines. One-half of the respondents considered at least 10 of 15 proposed criteria to decide patient discharge. ICUs in central referral hospitals used fewer criteria than community and private hospitals. The availability of intermediate care units was significantly greater in university hospitals. The ICU director's level of experience was not associated with the number of criteria used. In the five clinical scenarios there was wide variation in discharge decision. CONCLUSIONS: Our data indicate that there is marked heterogeneity in ICUs discharge practices, and that discharge decisions may be influenced by institutional factors. University teaching hospitals had more intermediate care facilities available. Written discharge guidelines were not widely used. PMID- 16249929 TI - Diphtheria, tetanus and pertussis antibodies in 10-year-old children before and after a booster dose of three toxoids: implications for the timing of a booster dose. AB - In an open study, 502 10-year-old children, who had received primary vaccination against diphtheria and tetanus in infancy and had varying histories of pertussis disease and vaccination, were vaccinated with diphtheria-tetanus vaccine (DT) alone or with the addition of 20 microg or 40 microg of pertussis toxoid. Diphtheria toxin neutralising antibodies, pertussis toxin IgG and tetanus toxoid IgG antibodies were measured before and 1 month after the booster. All toxoids were highly immunogenic. In pertussis toxoid recipients, median levels of pertussis toxin IgG increased to 16.5 U/ml (DTaP20) and to 36 U/ml (DTaP40) in children with non-detectable (<1 U/ml) antibodies before vaccination and to >400 U/ml in children (both DTaP20 and DTaP40) with detectable antibodies before vaccination. A total of 60 children (12%) with non-detectable (<0.01 IU/ml) diphtheria antibodies and 36 children (7%) with non-detectable (<0.01 IU/ml) tetanus antibodies before the booster had lower median antibody concentrations post-vaccination than children with detectable antibodies before the booster (diphtheria: 5.12 vs. 20.48 IU/ml; tetanus: 4.0 vs. 10.0 IU/ml). There were no differences in diphtheria and tetanus antibodies after vaccination between children who did and did not receive pertussis toxoid. CONCLUSION: 10-year-old children with non-detectable diphtheria and tetanus antibodies before the booster had lower post-vaccination antibodies than those with detectable antibodies before the booster indicating a poor immunological memory. Addition of pertussis toxoid to diphtheria-tetanus vaccine did not affect the antibody responses to diphtheria and tetanus toxoids when the three toxoids were combined as a booster. Even though immunity to diphtheria and tetanus was only estimated by surrogate markers (serum antitoxin antibodies) the results indicate that a lower age for the booster dose of diphtheria-tetanus vaccine or diphtheria-tetanus acellular pertussis vaccine should be considered. PMID- 16249930 TI - Prognostic utility of the semi-quantitative procalcitonin test, neutrophil count and C-reactive protein in meningococcal infection in children. AB - The aim was to determine whether semi-quantitative procalcitonin (PCT-Q) measurements on admission can identify the severity of meningococcal infection in children. A total of 65 children (mean age 2.4 years) with meningococcal disease were included in a prospective study. All patients were treated with antibiotics, rehydration, inotropic drugs and mechanical ventilation if presenting with shock or respiratory failure. On admission, blood was drawn for routine laboratory analyses including absolute neutrophil count (NC), C-reactive protein (CRP) and PCT-Q (immunochromatographic test). A total of 33 patients presented with septic shock on admission of whom 18 developed multiple organ dysfunction syndrome (MODS) and 9 died. Forty-three patients showed a very high PCT-Q level (>or=10 ng/ml), 12 showed values between 2-9.9 ng/ml and the remaining 10 patients showed PCT-Q levels<2 ng/ml. All patients with a PCT-Q level<10 ng/ml survived, whereas all those who developed MODS or died had PCT-Q levels>or=10 ng/ml. Receiver operator curve analysis showed that PCT-Q and NC had a high predictive value for MODS and death. PCT-Q showed a sensitivity of 100%, a negative predictive value of 100% and a negative likelihood ratio of 0.0 for MODS and death. CONCLUSION: semi-quantitative procalcitonin levels under 10 ng/ml predict good outcome of children with meningococcal infection. It is a highly sensitive method to identify patients with an increased risk of multiple organ dysfunction syndrome or death. PMID- 16249931 TI - Reduction of excessive height in boys by bilateral percutaneous epiphysiodesis around the knee. AB - In this study, the results of percutaneous epiphysiodesis as a surgical method to decrease final height is described in 15 boys with a predicted final height of more than 205 cm. A total of 17 boys with a height prediction between 195 and 209 cm without treatment were included as controls. The study period was from 1995 2002 and patients were followed for a mean period of 3.9 years (range 2.3-6.5 years) after surgery; controls were followed for 8.3 years (range 2.0-12.1 years). Final height in the treated boys was 203.6 cm (range 195.5-214.5 cm) compared to the predicted height of 210.6 cm (range 205.7-222.7 cm). The reduction in final height versus the predicted height was 7 cm and ranged between 1.2 and 13.8 cm. Final height in the control boys was 199.9 cm (range 191.3-206.7 cm). No significant side-effects of epiphysiodesis were observed. Besides final height reduction, epiphysiodesis resulted in normalisation of body proportions, expressed as the subischial leg length/sitting height ratio. This ratio in the operated patients at final height was 0.96 (range 0.90-1.01) and in the controls 0.94 (range 0.88-1.03). CONCLUSION: Epiphysiodesis can be advised as a method to decrease final height in boys with predicted tall stature. An additional advantage of this method is a normalisation of body proportions. PMID- 16249933 TI - Can economic evaluations be made more transferable? AB - Several commentators have identified the lack of generalisability and transferability of economic evaluation results. The aims of this study were: (a) to develop a checklist to assess the level of generalisability and transferability of economic evaluations; (b) to assess the generalisability and transferability of economic evaluations between the UK and France using the checklist; (c) to identify reasons for any lack of transferability and generalisability; (d) to assess how the transferability and generalisability of economic evaluations can be improved; and (e) to outline ways in which databases of economic evaluations and journals can assist in this area. The checklist was developed using previous work and the templates of the NHS EED and CODECS databases. A sub-checklist of essential items was then derived. Validation of the two checklists was undertaken with Health Economists participating in the EURONHEED project. Economic evaluations involving the UK and France were then located and assessed using the checklist. A summary score for each study was calculated based on the percentage of correctly reported (applicable) points, and the results in the empirical analysis compared to identify differences. The extended checklist includes 42 items, and the sub-checklist 16 items. Twenty-five economic evaluations met the inclusion criteria for the empirical analysis. In the extended checklist the mean score was 66.9+/-13.6%. The results for the sub checklist were very similar. The analysis revealed that costing, assessments of generalisability by the author(s), assessment of data variability, discounting, study population, and the reporting of effectiveness are areas that need more attention. Differences in cost-effectiveness results are often accounted for by price or organisational differences. The developed checklists are useful in assessing the generalisability and transferability of economic evaluations. In order to improve the generalisability and transferability of economic evaluations authors need to be more explicit and detailed in describing and reporting their studies. If they are to provide added value to their users, international databases of economic evaluations should systematically assess the generalisability and transferability of studies. Further research is in progress on producing a weighted version of the checklist. PMID- 16249932 TI - Impaired ability to increase water excretion in mice lacking the taurine transporter gene TAUT. AB - Cellular taurine uptake or release counteracts alterations of cell volume. Na+ coupled taurine transporter TAUT mediates concentrative cellular uptake of taurine. Inhibition of vasopressin secretion by hypotonicity may involve taurine release from glial cells of supraoptic nucleus. We compared renal function of mice lacking TAUT (taut-/-) and wild-type littermates (taut+/+). We observed renal taurine loss and subsequent hypotaurinemia in taut-/- mice. With free access to water, plasma and urine osmolality, urinary flow rate as well as urinary excretion and plasma concentrations of Na+ and K+ were similar in taut-/- and taut+/+ mice, whereas plasma concentrations of urea were enhanced in taut-/- mice. An oral water load (1 ml/16 g body weight) induced a similar diuresis in both genotypes. Repeating the oral water load immediately after normalization of urine flow rate, however, resulted in delayed diuresis and higher urinary vasopressin/creatinine ratios in taut-/- mice. In comparison, the repeated diuretic response to vasopressin V2 receptor blockade was not different between genotypes. Water deprivation for 36 h led to similar antidiuresis and increases of urinary osmolality in both genotypes. Upon free access to water after deprivation, taut-/- mice continued to concentrate urine up to 6 days, while taut+/+ mice rapidly returned to normal urinary osmolality. Urinary vasopressin/creatinine ratios and plasma aldosterone concentrations were not different under basal conditions but were significantly higher in taut-/- mice than in taut+/+ mice at 6 days after water deprivation. In conclusion, taut-/- mice suffer from renal taurine loss and impaired ability to lower urine osmolality and to increase urinary water excretion. The latter defect could reside extrarenally and result from a role of taurine in the suppression of vasopressin release which may be attenuated in taut-/- mice. PMID- 16249934 TI - Effect of oxygen limitation on the in vitro antimicrobial susceptibility of clinical isolates of Pseudomonas aeruginosa grown planktonically and as biofilms. AB - Pseudomonas aeruginosa, the predominant causative pathogen of chronic lung infection in patients with cystic fibrosis, may grow under anaerobic conditions as biofilms in the lungs of cystic fibrosis patients. To determine if growth under anaerobic conditions affects the antimicrobial susceptibility of P. aeruginosa, the susceptibility of clinical isolates of P. aeruginosa grown planktonically and as biofilms to a range of antibiotics was determined. Growth under anaerobic conditions did not reduce the ability of ceftazidime, meropenem, aztreonam, piperacillin, or piperacillin/tazobactam to inhibit planktonic growth, with MIC50 values for these antibiotics remaining unchanged or decreasing. However, tobramycin was less effective at inhibiting planktonic bacterial growth under anaerobic conditions, with the MIC50 of tobramycin increasing twofold. Growth under anaerobic conditions also decreased the bactericidal activity of tobramycin, with the MBC50 of tobramycin increasing fourfold. The killing kinetics of tobramycin was also examined under aerobic and anaerobic conditions for selected isolates. When isolates 6A and 12A were grown aerobically, concentration-dependent decreases in total viable count were apparent with tobramycin. In contrast, when these isolates were grown anaerobically, tobramycin at the same concentrations did not decrease the total viable count. When isolates were grown as biofilms under both aerobic and anaerobic conditions, isolate- and concentration-dependent differences in killing of the biofilms by tobramycin were apparent. However, tobramycin at concentrations up to 128 mg/l was unable to eradicate biofilms of any of the isolates tested, whether biofilms were grown aerobically or anaerobically. These results show that oxygen limitation may reduce, in a strain-dependent manner, the susceptibility to tobramycin of P. aeruginosa grown planktonically and as biofilms. PMID- 16249935 TI - Molecular modeling of the GABA(C) receptor ligand-binding domain. AB - We have constructed a molecular model of the ligand-binding domain of the GABA(C) receptor, which is a member of the Cys-loop ligand-gated ion channel family. The extracellular domains of these receptors share similar sequence homology (20%) with Limnaea acetylcholine-binding protein for which an X-ray crystal structure is available. We used this structure as a template for homology modeling of the GABA(C) receptor extracellular domain using FUGUE and MODELLER software. FlexX was then used to dock GABA into the receptor ligand-binding site, resulting in three alternative energetically favorable orientations. Residues located no more than 5 A from the docked GABA were identified for each model; of these, three were found to be common to all models with 14 others present only in certain models. Using data from experimental studies, we propose that the most likely orientation of GABA is with its amine close to Y198, and its carboxylate close to R104. These studies have therefore provided a model of the ligand-binding domain, which will be useful for both GABA(C) and GABA(A) receptor studies, and have also yielded an experimentally testable hypothesis of the location of GABA in the binding pocket. [Figure: see text]. PMID- 16249936 TI - QSTR with extended topochemical atom (ETA) indices. VI. Acute toxicity of benzene derivatives to tadpoles (Rana japonica). AB - structure-toxicity relationship (QSTR) studies have proved to be a valuable approach in research on the toxicity of organic chemicals for ranking chemical substances with respect to their potential hazardous effects on living systems. With this background, we have modeled here the acute lethal toxicity of 51 benzene derivatives with recently introduced extended topochemical atom (ETA) indices [Roy and Ghosh, Internet Electron J Mol Des 2:599-620 (2003)]. We also compared the ETA relations with non-ETA models derived from different topological indices (Wiener W, Balaban J, flexibility index, Hosoya Z, Zagreb, molecular connectivity indices, E-state indices and kappa shape indices) and physicochemical parameters (AlogP98, MolRef,H_bond_donor and H_bond_acceptor). Genetic function approximation (GFA) and factor analysis (FA) were used as the data-preprocessing steps for the development of final multiple linear regression (MLR) equations. Principal-component regression analysis (PCRA) was also used to extract the total information from the ETA/non-ETA/combined matrices. All the models developed were cross-validated using leave-one-out (LOO) and leave-many out techniques. The summary of the statistics of the best models is as follows: (1) FA-MLR: ETA model- Q 2 (LOO)=0.852, R 2=0.894; non-ETA model- Q 2=0.782, R 2=0.835; ETA + non-ETA model-Q 2 =0.815, R 2=0.859. (2) GFA-MLR: ETA model-Q 2 =0.847, R 2=0.915; non-ETA model-Q 2 =0.863, R 2=0.898; ETA + non-ETA model-Q 2 =0.859, R 2=0.893. 3. PCRA: ETA model-Q 2 =0.864, R 2=0.901; non-ETA model- Q 2=0.866, R 2=0.922; ETA + non-ETA model-Q 2=0.846, R 2=0.890. The statistical quality of the ETA models is comparable to that of non-ETA models. Again, use of non-ETA descriptors in addition to ETA descriptors does not increase the statistical acceptance of the relations significantly. The predictive potential of these models was better than that of the previously reported models using physicochemical parameters [Huang et al., Chemosphere 53:963-970 (2003)]. The relations from ETA descriptors suggest a parabolic dependence of the toxicity on molecular size. Furthermore, the toxicity increases with functionality contribution of chloro substituent and decreases with those of methoxy, hydroxy, carboxy and amino groups. This study suggests that ETA parameters are sufficiently rich in chemical information to encode the structural features that contribute significantly to the acute toxicity of benzene derivatives to Rana japonica. PMID- 16249937 TI - Ab initio and DFT study on the electrophilic addition of bromine to endo tricyclo[3.2.1.0(2,4)]oct-6-ene. AB - Full geometric optimization of endo-tricyclo[3.2.1.0(2,4)]oct-6-ene (endo-TCO) by ab initio and DFT methods allowed us to investigate the structure of the molecule. The double bond is endo-pyramidalized and its two faces are no longer found to be equivalent. The exo face of the double bond has regions with far more electron density (q(i,HOMO)) and more negative electrostatic potential. The endo TCO-Br2 system was investigated at the B3LYP/6-311+G** level and the endo TCO...Br2(exo) molecular complex was found to be relatively more stable than the endo-TCO...Br2(endo) complex. The cationic intermediates of the reaction were studied by ab initio and DFT methods. The bridged exo-bromonium cation(I) is relatively more stable than the endo-bromonium cation(II). An absolute exo-facial selectivity should be observed in the addition reaction of Br2 to endo-TCO, which is caused by steric and electronic factors. The nonclassical rearranged cation IV was found to be the most stable ion among the cationic intermediates and the ionic addition occurs via the formation of this cation. The mechanism of the addition reaction is also discussed. PMID- 16249940 TI - Massa intermedia: mental development. PMID- 16249938 TI - DNA methylation and epigenetic inheritance during plant gametogenesis. AB - In plants, newly acquired epigenetic states of transcriptional gene activity are readily transmitted to the progeny. This is in contrast to mammals, where only rare cases of transgenerational inheritance of new epigenetic traits have been reported (FASEB J 12:949-957, 1998; Nat Genet 23:314-318, 1999; Proc Natl Acad Sci U S A 100:2538-2543, 2003). Epigenetic inheritance in plants seems to rely on cytosine methylation maintained through meiosis and postmeiotic mitoses, giving rise to gametophytes. In particular, maintenance of CpG methylation ((m)CpG) appears to play a central role, guiding the distribution of other epigenetic signals such as histone H3 methylation and non-CpG DNA methylation. The evolutionarily conserved DNA methyltransferase MET1 is responsible for copying (m)CpG patterns through DNA replication in the gametophytic phase. The importance of gametophytic MET1 activity is illustrated by the phenotypes of met1 mutants that are severely compromised in the accuracy of epigenetic inheritance during gametogenesis. This includes elimination of imprinting at paternally silent loci such as FWA or MEDEA (MEA). The importance of DNA methylation in gametophytic imprinting has been reinforced by the discovery of DEMETER (DME), encoding putative DNA glycosylase involved in the removal of (m)C. DME opposes transcriptional silencing associated with imprinting activities of the MEA/FIE polycomb group complex. PMID- 16249941 TI - Decompressive craniectomy for traumatic brain injury: when is it too late? PMID- 16249942 TI - Surgical anatomy of the medial collateral ligament and the posteromedial capsule of the knee. AB - The supporting structures on the medial side of the knee consist of a superficial fascial layer (I), a deep capsular layer (III) with the deep medial collateral ligament in it and in between the superficial collateral ligament (layer II). The attachment sites of the different ligaments and the functions of the various ligamentous structures are described and suggestions for surgical repair are given. PMID- 16249943 TI - Self-reported and serum cotinine-validated smoking in pregnant women in Estonia. AB - OBJECTIVES: Although widely used in epidemiological studies, self-report has been shown to underestimate the prevalence of smoking among pregnant women. Objectives of this study were to examine the discrepancy between self-reported and cotinine validated smoking status, and the sociodemographic characteristics associated with the misclassification of real smoking status among pregnant women in Tallinn, the capital of Estonia. METHODS: Serum cotinine assays were performed on a subsample (n = 1360) of the pregnant women, who had participated in a recent study of human papillomavirus type 16 (HPV-16) seroprevalence in Estonia. In the present study, serum concentrations > or = 15 ng/ml were used to distinguish current smokers from nonsmokers. The serum-validated smoking level was compared with the self-reported level in the records of the Estonian Medical Birth Registry. For the group of self-reported non-smokers, the differences between the cotinine-validated smokers and the cotinine-validated nonsmokers, with respect to their sociodemographic characteristics (age, ethnicity, educational level, employment status, marital status, parity), were estimated by logistic regression. RESULTS: Of 1239 women who reported being nonsmokers, 259 (20.9%) had serum cotinine levels > or = 15 ng/ml, and can be regarded as current smokers. Among self-reported nonsmokers, nondisclosure of current smoking was significantly more frequent in non-Estonian, less educated, socially inactive, cohabiting and multiparous women. CONCLUSIONS: Self-reported data on smoking in pregnant women underestimates the real smoking prevalence in Estonia. Maternal unwillingness to declare smoking during pregnancy needs to be taken into account in the practice of maternal and child health to better target prenatal smoking cessation interventions. PMID- 16249944 TI - Pregnancy intendedness, maternal psychosocial factors and preterm birth. AB - OBJECTIVES: This study examined associations between reported pregnancy intendedness and several maternal psychosocial factors in relation to preterm birth (<37 weeks' completed gestation). METHODS: Women were recruited into a prospective cohort study between the 24th and 29th weeks of pregnancy in central North Carolina from 1996 to 2000. Prior to delivery, participants responded to questions about pregnancy intendedness, life events impacts, depressive symptoms, and coping style. RESULTS: Women who reported not intending their pregnancy had increased odds of reporting low, medium and high levels of perceived stress during pregnancy (OR = 1.4 [95% CI: 1.1, 1.9], OR = 2.2 [95% CI: 1.7, 2.8], and OR = 3.4 [95% CI: 2.6, 4.5], respectively, relative to very low), medium and high levels of depressive symptoms (OR = 2.2 [95% CI: 1.8, 2.9] and OR = 3.1 [95% CI: 2.4, 3.9], respectively), and medium and high levels of several coping styles. Reporting not intending the pregnancy was not associated with increased risk of preterm birth (Risk Ratio [RR] = 1.0, 95% CI: 0.8, 1.1), but reporting the highest quartile of perceived stress (RR = 1.6, 95% CI: 1.1, 2.3) and the highest tertile of distancing coping style (compared with lowest quartile) was associated with preterm birth (RR = 1.4, 95% CI: 1.1, 1.9). Interactions between pregnancy intendedness and the psychosocial variables perceived stress, depression or coping style did not modify the psychosocial variable's associations with preterm birth. CONCLUSIONS: Pregnancy intendedness remains an important concept in the reproductive health literature integrally tied to indicators of maternal mental health, but not necessarily to pregnancy outcomes. PMID- 16249945 TI - Traditional healers for HIV/AIDS prevention and family planning, Kiboga District, Uganda: evaluation of a program to improve practices. AB - In the face of ongoing epidemics of HIV/AIDS and STI, high demand for family planning, and limited resources, traditional healers may be under-utilized providers of reproductive health education in rural sub-Saharan Africa. We implemented a training program in HIV prevention and family planning methods for healers in the Kiboga district of Uganda and evaluated the program's impact on healers' clinical practice and the diffusion of information to their female clients. Of 46 healers recruited, 30 (65%) completed a pre- and post-training interview. Following training, traditional healers increased discussions of family planning with their clients. Of 84 female clients recruited, 44 (52%) completed the interview before and after the training for healers. Female clients corroborated that they increased discussions of family planning with their healers, as well as discussions about HIV/AIDS. Both healers and their female clients were more likely to make a connection between family planning, condom use, and HIV prevention after the training compared to before the training. Findings provide evidence that traditional healers in a rural area of Uganda can successfully adapt HIV prevention messages and family planning information into their clinical practices. PMID- 16249946 TI - Condom attitudes and behaviors among injection drug users participating in California syringe exchange programs. AB - This study examined condom attitudes, preferences, barriers, and use among a sample of 550 injection drug using clients of syringe exchange programs in California. In multivariate analyses, positive attitudes toward condoms were significantly associated with consistent condom use for vaginal, anal, and oral sex in the past six months, beyond the effects of confounding socio-demographic and HIV risk variables. Participants commonly cited partner-related barriers to condom use, such as reluctance to use condoms with steady partners (34%). Almost a quarter of the sample cited dislike of condoms (e.g., because of pleasure reduction). In addition, a third of respondents stated specific preferences regarding condom brands, sensitivity, sizes, and textures. Interventions that increase awareness about positive aspects of condom use and sexual risk from steady partners may be successful in increasing condom use among injection drug users. PMID- 16249947 TI - Genetic knowledge and collective identity. PMID- 16249948 TI - "Bioethics in action" and human population genetics research. AB - Recent disputes about human population genetics research have been provoked by the field's political vulnerability (the historic imbalance of power between the geneticists and the people they study) and conceptual vulnerability (the mismatch between scientific and popular understandings of the genetic basis of collective identity). The small, isolated groups often studied by this science are now mobilizing themselves as political subjects, pressing sovereignty claims, and demanding control over the direction and interpretation of research. Negotiations between the geneticists and the people asked to donate DNA have resulted not only in explicit bioethics protocols but also in diffuse anxiety over the incommensurability between expert and non-expert views about genetic evidence for identity claims. This article compares two disputes over genetics research: the Human Genome Diversity Project and the use of genetics to prove identity claims among the Melungeons of Tennessee. The case studies illustrate "bioethics in action": how particular controversies and interests drive the production of bioethics discourses and techniques (such as informed consent protocols). They also illustrate some limits on the usual apparatus of bioethics in overcoming this science's multiple vulnerabilities. PMID- 16249949 TI - The multiple meanings of Jewish genes. AB - This article addresses contemporary social challenges created by new genetic research on Jews and by Jews, and its implications for the meanings of Jewish identity, on both the individual and the collective levels. The article begins with a brief overview of selective genetic studies of Jewish populations and the controversies they have generated. It continues with an examination of the emerging field of Jewish genetic demography, which employs genetic tests to identify lineages, claim kin, and support Jewish historical and political claims. Here the article explores how Jewish genetic demographers interpret genetic studies to reinforce oral tradition and Biblical prophecy about the origins of the Jews and their experience in the Diaspora. This research is then juxtaposed with debates that emerge from contemporary rabbinic deliberations over the appropriate uses of new reproductive technologies, debates that, contrary to the assertions of Jewish genetic demographers, suggest genes are believed to possess limited ability to confer or create Jewishness in the traditional rabbinic imagination. In the final section of this article, a debate is staged about contemporary biomedical practices that allow for the exchange and transfer of body parts and bodily substances, as a strategy for challenging genetic notions of Jewish identity. PMID- 16249950 TI - Genetics, history, and identity: the case of the Bene Israel and the Lemba. AB - The paper examines the impact of genetic research on the religious identity of the Bene Israel Indian Jewish community and the Lemba Judaising group of southern Africa. It demonstrates how DNA tests which happened to support the possibility of the communities' legends of origin affected their self-perception, the way they are viewed by their neighbors, and their image in the West. It is argued that in both cases what accounted most for the Bene Israel and Lemba responses to the tests was the way the results were portrayed in the mass media, the history of the development of Judaism in their communities, and the local realities. PMID- 16249951 TI - Genetic genealogy: the Woodson family's experience. AB - In 1998, Foster and colleagues published the results of a genetic study intended to test whether Thomas Jefferson could have fathered any of Sally Hemings' children. They found that the Jefferson Y chromosome haplotype matched that of a descendant of Hemings' youngest child, but not that of the descendants of the eldest son, Thomas Woodson. The Woodson descendants were shocked by the study's finding, which disagreed with their family oral history. They were suspicious of the study conclusions because of the methods used in recruiting participants for the study and the manner in which they learned of the results. The Woodsons' experience as participants in one of the first examples of genetic genealogy illustrates several issues that both geneticists and amateur genetic genealogists will face in studies of this kind. Misperceptions about the relationship between biology and race, and group genetics in general, can make the interpretation of genetic data difficult. Continuing collaborations between the media and the scientific community will help the public to better understand the risks as well as the benefits of genetic genealogy. Researchers must decide prior to beginning their research what role the human subjects will play in the study and when they will be notified of the study's conclusions. Amateur genetic genealogists should anticipate unexpected outcomes, such as the identification of nonpaternity, to minimize any harmful effects to study participants. Although modern genetic methods provide a powerful new tool for genealogical study, they cannot resolve all genealogical issues, as this study shows, and can involve unanticipated risks to the participants. PMID- 16249952 TI - Prologue to special issue "visual electrophysiology in drug development". PMID- 16249953 TI - An overview of drug development with special emphasis on the role of visual electrophysiological testing. AB - Visual electrophysiological techniques, such as electroretinography (ERG) and visual evoked potentials (VEP) can provide useful information on the safety, efficacy, and proper dosing of chemical entities under development as new drug therapies. During post-marketing safety surveillance, a variety of visual electrophysiological measures can be used to objectively assess and document individual patient response to ophthalmic drugs and ocular or visual system side effects of non-ophthalmic drugs. In this paper, the discovery, exploratory development, full-development and post-marketing stages of drug development are briefly outlined. The potential role of visual electrophysiological techniques in each of these stages is described and discussed. PMID- 16249954 TI - Contribution to ischemic injury of rat optic nerves by intracellular sodium overload. AB - Ischemic insult to axons of retinal ganglion cells (RGCs) is believed to contribute significantly to preferential loss of RGCs in glaucoma. In this study, we characterized the role of intracellular Na(+) overload in ischemic injury of acutely isolated rat optic nerves by evaluating electrically elicited compound action potentials (CAPs) from the optic nerves. Under control conditions, robust and stable CAPs can be recorded for more than 5 h. One hour of oxygen and glucose deprivation (OGD) that simulates ischemia, virtually eliminated the CAP. Upon returning to control conditions, the CAP gradually recovered. Maximum recovery (35% of control) was obtained by 1 h after returning to normal oxygenated Ringer. When a rapidly reversible Na(+) channel blocker, that completely blocked the CAP under control conditions, was present during OGD, the recovery of the CAP was significantly enhanced to 65% of control. When the Na(+) was replaced with either choline or Li(+) in the Ringer during OGD, CAP recovery was significantly enhanced (65-70% of control). Removing Ca(++) from the Ringer (plus 5 mM EGTA) provided even better preservation of the CAP following OGD (90% of control). Our results are consistent with the hypothesis that intracellular Na(+) overload appears to play a significant role in ischemic injury of optic nerves. This Na(+) overload may depend at least partially upon Ca(++) influx from the extracellular space. PMID- 16249955 TI - Diltiazem-induced neuroprotection in glutamate excitotoxicity and ischemic insult of retinal neurons. AB - PURPOSE: Cell death is often related to an abnormal increase in Ca(2+) flux. In the retina, Ca(2+) channels are mainly from the L-type that do not inactivate with time. Under excitotoxic and ischemic conditions, their continuous activation may therefore contribute significantly to the lethal Ca(2+) influx. To assess this hypothesis, the Ca(2+) channel blocker, diltiazem, was applied in excitotoxic and ischemic conditions. METHODS: To induce excitotoxicity, retinal cell cultures from newborn rats were incubated with glutamate. The toxicity of glutamate was quantified by neuronal immunostaining with an antibody directed against the neuron specific enolase. Glutamate receptor function in vitro was assessed in pig retinal cell cultures by patch clamp recording. Retinal ischemia was induced by raising the intraocular pressure in adult rats. Retinal cell loss was quantified on retinal sections by measuring nuclear cell densities. RESULTS: In retinal cell culture, glutamate application induced a major cell loss. This cell loss was attributed to glutamate excitotoxicity because glutamate receptor blockers like MK-801 and CNQX increased significantly neuronal survival. MK-801 and CNQX, which block NMDA and AMPA/Kainate receptors, respectively, had additive effects. Expression of AMPA/Kainate glutamate receptors in mixed adult retinal cell cultures was attested by patch clamp recording. In newborn rat retinal culture, glutamate excitotoxicity was significantly reduced by addition of the L type Ca(2+) channel blocker, diltiazem. In in vivo experiments, the increase in ocular pressure induced a decrease in cell number in the inner nuclear and ganglion cell layers. When animals received diltiazem injections, the ischemic treatment induced a less severe reduction in retinal cells; this neuroprotection was statistically significant in the ganglion cell layer. CONCLUSION: These results are consistent with previous studies suggesting that Ca(2+) channel activation contributes to retinal cell death following either glutamate excitotoxicity or retinal ischemia. Under both conditions, the L-type Ca(2+) channel blocker, diltiazem, can limit cell death. These results extend the potential application of diltiazem in retinal neuroprotection to retinal pathologies involving glutamate excitotoxicity and ischemia. PMID- 16249957 TI - Recommendations for a toxicological screening ERG procedure in laboratory animals. AB - Electroretinography, using laboratory animals, is a commonly used technique for determining the retinal toxicity of chemical agents. In this paper, guidelines for performing this test are provided. The physiologic basis for visual testing is presented with attention to inter-species differences. Technical aspects of animal recordings are reviewed, including animal preparation, stimulation, signal conditioning, recording and data analysis. Finally, suggested protocols for recording in diurnal and nocturnal species are presented. PMID- 16249956 TI - Retinal function assessed by ERG before and after induction of ocular aspergillosis and treatment by the anti-fungal, micafungin, in rabbits. AB - This study was conducted to evaluate the effectiveness of a new antifungal drug, micafungin, and standard antifungal drugs against endophthalmitis induced in a rabbit by intravitreal injection of Aspergillus fumigatus, an important fungal pathogen. Effectiveness was evaluated by the preservation of b-wave amplitude at 72 h after injection of the fungus relative to the b-wave amplitude at baseline before any intravitreal injections. A 0.06 ml inoculum of 10(6) conidia of A. fumigatus was injected into the vitreous of the right eye of all rabbits; and, 12 h later, a 0.06 ml solution containing one of 3 antifungal drugs or saline was injected into the vitreous of both eyes. All three antifungal drugs produced significant b-wave preservation at 72 h in infected eyes compared to that in infected eyes receiving saline injections. There was no statistically significant difference between the effects of micafungin and amphotericin B in the right eyes with fungal endophthalmitis, and both produced significantly more preservation of b-wave amplitude than voriconazole. Amphotericin B, but neither micafungin nor voriconazole produced significant reduction of the b-wave amplitude in the left eyes. PMID- 16249958 TI - The photopic ERG of the albino guinea pig (Cavia porcellus): a model of the human photopic ERG. AB - Altricial rodents such as rats and mice are probably the most widely used animal model in the electroretinogram (ERG) literature. However, while the scotopic responses of these rodents share obvious similarities with that of humans, their photopic electroretinograms are strikingly different. For instance, the photopic ERGs of rats and mice include, when measurable, a minimal a-wave, while the b wave is of much larger amplitude than that of humans. The purpose of this study is to present the albino guinea pig which is like humans, is a precocial animal, and is a better rodent model of the human photopic ERG. In order to investigate the above, photopic electroretinograms and oscillatory potentials, obtained from guinea pigs and human subjects, were compared. Furthermore, in a subset of animals we injected, intravitreally, selective blockers of the ON- (L-2-amino-4 phosphonobutyric acid: L-AP-4; 10 mM) or OFF- (kynurenic acid: KYN; 50 mM) retinal pathways in order to mimic similar retinal disorders found in human. Based on our results, we believe that, compared to rats and mice, the photopic (cone-mediated) ERG of the guinea pig clearly represents a superior rodent model of the human photopic ERG. PMID- 16249959 TI - Repeated spike exposure to the insecticide chlorpyrifos interferes with the recovery of visual sensitivity in rats. AB - Reports from Japan and India and data submissions to the US EPA indicate that exposure to cholinesterase (ChE)-inhibiting organophosphorous insecticides (OP) can produce ocular toxicity, in particular long-lasting changes in retinal physiology and anatomy. We have examined the effects of a 1 year dietary exposure to the OP chlorpyrifos (CPF) on retinal structure and function. Adult male Long Evans rats were fed CPF in their diet at the rate of 0, 1 (low), or 5 (high) mg/kg body weight/day. In addition, half of each feeding group received an oral (spike) dose of CPF in corn oil (45 mg/kg) or corn oil (VEH) alone every 2 months, resulting in six exposure groups: Control-VEH, Control-CPF, Low-VEH, Low CPF, High-VEH, and High-CPF. Dark-adapted electroretinograms (ERG) were measured 3-5 months (n= 15-18/group) after the completion of dosing. There were no significant differences between dose or spike groups in a-wave, b-wave, or oscillatory potential amplitudes or implicit times. In addition, the time course of dark adaptation were measured in a subset of these rats (6-8/group) eight months after the completion of dosing by determining the flash intensity needed to elicit a 40 microV b-wave at selected intervals after bleaching 90% of the photopigment. Rats receiving the episodic oral spike of CPF showed a slowed recovery of dark-adapted sensitivity compared to rats receiving the corn oil VEH across chronic dosing conditions. No effects were seen on retinal morphology. This result suggests that episodic high dose exposures to CPF may result in altered retinal function. This effect, akin to effects seen in aging humans and humans exposed to other ChE-inhibiting compounds, may reflect alterations in the photoreceptors and retinal pigment epithelium (RPE) complex necessary for regenerating photopigment. PMID- 16249960 TI - Electrophysiological assessment of glaucomatous visual dysfunction during treatment with cytidine-5'-diphosphocholine (citicoline): a study of 8 years of follow-up. AB - In this study we assessed, by simultaneous recordings of visual evoked potentials (VEPs) and pattern-electroretinograms (PERGs), the effects cytidine-5' diphosphocholine (citicoline) on retinal function and/or visual cortical responses in glaucoma patients. Thirty glaucoma patients were randomly divided into two age-matched groups: patients in group GC (15 patients) were treated with citicoline (1,000 mg/die intramuscularly) for 2 months; patients in group GP (15 patients) were treated with placebo for 2 months. After 4 months of wash-out (month 6), GC patients underwent a further 2-month period of citicoline treatment (months 7-8) followed by another 4-month period of wash-out (months 9-12). In GP patients the wash-out was extended for a further 6 months (months 7-12). During the following 13-96 months, GC patients received additional 2-month periods of treatment with citicoline (each period followed by 4 months of wash-out) for a total of 16 periods in 8 years. GP patients were also examined at months 24, 26, 48, 60, 72, 84 and 96. In GC patients the first two treatments with citicoline induced a significant (p <0.01) improvement of VEP and PERG parameters with respect to pre-treatment conditions. VEPs and PERGs recorded in GC patients after the first wash-out revealed that, although there was a worsening trend, the electrophysiological improvement was still maintained with respect to baseline conditions. The additional periods of citicoline treatment in GC patients during the subsequent 13-96 months induced a greater (p <0.01) improvement of VEP and PERG parameters with respect to pre-treatment conditions and when compared to GP patients. Thus, we observed that citicoline significantly improves retinal and cortical bioelectrical responses in glaucoma patients, suggesting a potential use of this substance in the medical treatment of glaucoma, as a complement to hypotensive therapy. PMID- 16249961 TI - Regional assessment of cone system function following uncomplicated retinal detachment surgery. AB - PURPOSE: To evaluate regional cone system function after uncomplicated retinal detachment (RD) surgery, by recording focal electroretinograms (FERGs) from the central and paracentral regions of the posterior pole. METHODS: FERGs in response to either a central (eccentricity: 0-2.25 deg) or a paracentral annular (2.25-9 deg) uniform field, presented on a light adapting background and sinusoidally flickered at 41 Hz (95% modulation depth, 93 cd/m(2) mean luminance) were recorded from 16 eyes (16 patients) 2 weeks to 420 months following uncomplicated RD surgery (encircling procedure with or without scleral buckling). Pre operatively, 10 out of 16 eyes had a macular RD. Mean time elapsed from onset of symptoms to surgery was 20 days (range: 5-90 days). Post-operatively, visual acuity ranged 0.1 to 1.0. Eight age-matched normal subjects served as controls. Amplitude and phase of the FERG fundamental harmonic were measured. RESULTS: Compared to control eyes, affected eyes' central and paracentral FERGs were on average reduced in amplitude (by 40% and 28%, respectively, p<0.01) and delayed in phase (by 70 and 100 degrees, respectively, p<0.01). Eyes with a macular RD did not differ in FERG amplitude or phase from eyes that had a peripheral RD. In individual affected eyes, central, but not paracentral FERG amplitudes were negatively correlated (p=0.05) with time elapsed from onset of symptoms to surgery. CONCLUSIONS: Central and paracentral cone system dysfunction may be detected even at lengthy time intervals from retinal reattachment, independent of the presence of a pre-operative macular RD. The extent of central loss appears to be inversely related to RD duration, in agreement with previous reflectometric findings on foveal cone photopigment density [Liem et al., 1994; Ophthalmology 10: 1945-51]. PMID- 16249964 TI - Effects of freshly neutralized aluminum on oxygen consumption by freshwater invertebrates. AB - The hypothesis tested in this study was if a pulse of precipitating aluminum (Al) at circumneutral pH covers the body of an invertebrate and, therefore, reduces the surface area available for respiration, organisms exposed to precipitating Al in an experimental system should consume less oxygen than organisms not exposed to aluminum. To test this hypothesis, experiments were conducted in the laboratory placing test organisms in biochemical oxygen demand (BOD) bottles containing a recently neutralized acidic, Al-enriched solution; conditions were meant to loosely mimic those of an acidic, Al-rich stream flowing into a larger, neutralizing receiving stream. The experiments suggested that freshly neutralized Al, i.e., Al in transition from ionic species in acidic waters to polymers or precipitating hydroxides after a rapid increase to pH > or = 6.8, impaired oxygen consumption by D. magna in a repeatable, dose-dependent fashion. Precipitating Al also impaired oxygen consumption by the perlid stoneflies Perlesta lagoi and Acroneuria abnormis, at the lower concentrations used, but higher concentrations resulted in oxygen consumption similar to that of controls. An ionoregulatory impairment response may explain this trend. Aluminum did not affect oxygen consumption by the larger, detritivorous stonefly, Pteronarcys pictetii. PMID- 16249963 TI - Epilepsy and medication effects on the pattern visual evoked potential. AB - Visual disruption in patients diagnosed with epilepsy may be attributable to either the disease itself or to the anti-epileptic drugs prescribed to control the seizures. Effects on visual function may be due to perturbations of the GABAergic neurotransmitter system, since deficits in GABAergic cortical interneurons have been hypothesized to underlie some forms of epilepsy, some anti epileptic medications increase cortical GABA levels, and GABAergic neural circuitry plays an important role in mediating the responses of cells in the visual cortex and retina. This paper characterizes the effects of epilepsy and epilepsy medications on the visual evoked response to patterned stimuli. Steady state visual evoked potentials (VEP) evoked by onset-offset modulation of high contrast sine-wave stimuli were measured in 24 control and 54 epileptic patients. Comparisons of VEP spectral amplitude as a function of spatial frequency were made between controls, complex partial, and generalized epilepsy groups. The effects of the GABA-active medication valproate were compared to those of carbamezepine. The amplitude of the fundamental (F1) component of the VEP was found to be sensitive to epilepsy type. Test subjects with generalized epilepsy had F1 spatial frequency-amplitude functions with peaks shifted to lower spatial frequencies relative to controls and test subjects with complex partial epilepsy. This shift may be due to reduced intracortical inhibition in the subjects with generalized epilepsy. The second harmonic component (F2) response was sensitive to medication effects. Complex partial epilepsy patients on VPA therapies showed reduced F2 response amplitude across spatial frequencies, consistent with previous findings that showed the F2 response is sensitive to GABA-ergic effects on transient components of the VEP. PMID- 16249962 TI - Ocular toxicity due to chloroquine and hydroxychloroquine: electrophysiological and visual function correlates. AB - Chloroquine (CQ) and hydroxycholorquine (HCQ) have been used widely for the treatment of rheumatoid arthritis and other similar inflammatory diseases since the early 50s. They remain the treatment of choice for many patients even today. Significant, either reversible or irreversible central visual loss associated with the drugs is very rare, but an important side effect that can warrant discontinuation of therapy. Early diagnosis of toxicity and evaluation of the visual function are, therefore, important parts of the treatment process. Various electrophysiological and psychophysical tests have been and are used for the detection, follow-up and prognosis of drug-associated central visual loss. A summary and comment on the tests, with emphasis on the use of more recently developed methods, such as the multifocal electroretinography (mfERG), are presented in this review. PMID- 16249965 TI - Relief of arsenate toxicity by Cd-stimulated phytochelatin synthesis in the green alga Chlamydomonas reinhardtii. AB - In most photosynthetic organisms, inorganic arsenic taken up into the cells inhibits photosynthesis and cellular growth. In a green alga, Chlamydomonas reinhardtii, 0.5 mM arsenate inhibited photosynthesis almost completely within 30 min. However, in cells acclimated with a sublethal concentration (0.05 to 0.1 mM) of Cd, the inhibition of photosynthesis at 30 min after the addition of arsenate was relieved by more than 50%. The concentrations of arsenic incorporated into the cells were not significantly different between the Cd-acclimated and the non acclimated cells. The Cd-acclimated cells accumulated Cd and synthesized phytochelatin (PC) peptides, which are known to play an important role in detoxification of heavy metals in plants. By the addition of an inhibitor of glutathione (an intermediate in the PC biosynthetic pathway) biosynthesis, buthionine sulfoximine, cells lost not only Cd tolerance but also arsenate tolerance. These results suggest that glutathione and/or PCs synthesized in Cd acclimated cells are involved in mechanisms of arsenate tolerance. PMID- 16249967 TI - Hypothesized kinetic models for describing the growth of globular and encrusting demosponges. AB - The marine sponges Dysidea avara and Chondrosia reniformis (globular forms) were cultured in the laboratory on a diet of viable Phaeodactylum tricornutum cells and dissolved nutrients (algae and fish powders). Our growth data were combined with literature data for Pseudosuberites andrewsi (a globular sponge) and for the encrusting sponges Oscarella lobularis, Hemimycale columella, and Crambe crambe. The suitability of three growth models-linear, exponential, and radial accretive for describing the growth of globular and encrusting sponges was assessed. Radial accretive growth was determined to be the best model to describe growth of both encrusting and globular sponges. Average growth rates of 0.051+/-0.016 and 0.019+/-0.003 mm/day (calculated as the increase of the radius of the sponge per day) were obtained experimentally for D. avara and C. reniformis, respectively. PMID- 16249966 TI - Production and characterization of a continuous embryonic cell line from sea bass (Dicentrarchus labrax L.). AB - Continuous cell lines represent an important tool both for biological studies and for their applications in marine biotechnology. In this article we describe the production and characterization of a continuous adherent cell line, named DLEC, derived from early embryos of the European sea bass Dicentrarchus labrax L. (Actinopterygii, Moronidae). Cells were obtained by disrupting 2- to 12-hour-old embryos and culturing resulting cells at 18 degrees C in RPMI medium containing 5% fetal calf serum (FCS) and 10% supernatant fraction of the embryo homogenate. After 8 weeks culture medium was replaced with Liebovitz's L15 medium containing 10% FCS and DLEC cells started proliferation. Subsequently, they were continuously cultured until the 50th passage without evident changes in their morphology. DLEC cells show a fibroblast-like shape and a modal chromosome number of 48, as do the wild-type cells; conversely the constant presence of six to nine meta-submetacentric elements in the karyotype (vs. zero to two in the wild-type) indicates the occurrence of chromosomal rearrangements during stabilization. DLEC cells are sensitive to substances known to induce differentiation of mammalian cells such as retinoic acid and phorbol esters. They have been transfected using liposomes with a commercial plasmid vector containing a reporter gene, thus suggesting a possible importance as an alternative expression system of recombinant vertebrate proteins in teleost cells. PMID- 16249969 TI - Multimedia article. Entirely thoracoscopic pneumonectomy using the prone position: a new technique. AB - BACKGROUND: Reports on video-assisted pneumonectomy have remained scarce, despite early demonstration of its technical feasibility. A totally videothoracoscopic pneumonectomy was first reported by Conlan and Sandor. The patient in this report was positioned in the full lateral position. In this video, we report a totally videothoracoscopic left-side pneumonectomy with the patient in prone position. METHODS: A 49-year-old man was admitted to our hospital for a bifocal cancer of the left lower lung lobe (LLL) and the cervical esophagus. The preoperative workup included a chest computed tomography (CT) scan showing a 3-cm mass of the laterobasal segment of the LLL, with retrotumoral atelectasis, lymph nodes smaller than 1 cm in diameter at the aortopulmonary window and under the carena, and finally posterolateral adherences between the parietal and the visceral pleura. Flexible bronchoscopy confirmed the presence of a bronchial tumor at the offspring of the apical bronchus of the LLL. Biopsy showed invasive adenocarcinoma, and a CT scan of the neck and head was significant for tumoral infiltration of the cervical esophagus and retropharyngeal space. Gastroscopy showed a stenosis of the cervical esophagus and hypopharynx. Biopsy showed spinocellular epithelioma, but CT scan of the abdomen and bone scintigraphy did not show metastatic disease. A position emission tomography (PET) scan confirmed the findings of the CT scan. Pneumonectomy and esophagectomy by thoracoscopy, laparoscopy, and cervicotomy were proposed. The purpose of this video is to show the details of the thoracoscopic technique with the patient in the prone position. RESULTS: After induction of general anesthesia, a double-lumen endotracheal tube was placed. The patient was subsequently placed and strapped in a prone position. The surgical team was placed to the left of the patient. A 10 mm trocar was placed in the seventh intercostal space on the posterior axillary line, and a 30 degrees angled videoscope was introduced. Three additional 5-mm trocars were placed at the same level in the 5th, 9th, and 11th intercostal spaces on the posterior axillary line. The mediastinal pleura was opened just ventral to the aorta. The first structure identified was the left main bronchus, which was dissected free and transected with a linear stapler (blue load). The aortopulmonary window became immediately visible. Clearance of this window's lymphoglandular tissue showed, bottom to top, the inferior pulmonary vein, the superior pulmonary vein, and the pulmonary artery. These vascular structures were carefully dissected free with the cautery hook and transected with a vascular linear stapler (white load). The lung was freed entirely tend placed in a retrieval bag for later transhiatal extraction during the laparoscopic phase of the esophagectomy. The intraoperative time for the pneumonectomy was 146 min, and intraoperative blood loss was 30 ml. The pathology report confirmed the presence of invasive, poorly differentiated adenocarcinoma. The bronchial section was free of tumor. One intrapulmonary lymphnode (N1) was positive, whereas all 10 N2 and N3 nodes harvested were free of disease. The tumor was thus staged as IIB (pT2N1Mx). The esophagetomy specimen showed fairly wide differentiated keratinizing of the spinocellular epithelioma with invasion of both pyriform sinuses and both sides of the glottis. CONCLUSIONS: First described by Cuschieri et al. in 1992, the prone position for thoracoscopy allows for a more direct approach to the aortopulmonary window under excellent visual and ergonomic circumstances. Dissection of the hilar larger vessels and performance of lymphnode sampling appear more straightforward because with this technique, the lung is kept out of harm's way, thanks to gravity. PMID- 16249970 TI - Does Expressed Emotion need to be understood within a more systemic framework? An examination of discrepancies in appraisals between patients diagnosed with schizophrenia and their relatives. AB - BACKGROUND: This study examined the relationship between discrepancies in beliefs about schizophrenia and Expressed Emotion (EE) in family dyads. METHOD: Illness beliefs were assessed in patients with a diagnosis of schizophrenia and their relatives (N=49 dyads). The degree and the direction of discrepancy were calculated, and comparisons were made between dyads in which the relative was rated as high EE and dyads in which the relative was rated as low EE. RESULTS: There was greater discrepancy between illness models of schizophrenia in dyads involving a high-EE relative than in dyads involving a low-EE relative. This difference was not accounted for by the differences in either relatives' or patients' beliefs alone. CONCLUSION: Further research is needed to understand EE in the context of discrepancies in beliefs between patients and relatives rather than focussing on relatives' beliefs alone. PMID- 16249972 TI - Peri and postoperative pulmonary complications among cirrhotic individuals. AB - Surgical intervention among cirrhotic individuals carries a high risk for peri and postoperative complications. We review the literature regarding the frequency and consequences of pulmonary complications in cases of cirrhosis. The experience with hepato-pulmonary syndrome and porto-pulmonary hypertension in liver transplant recipients is also presented. PMID- 16249971 TI - Generalised anxiety disorder in Singapore: prevalence, co-morbidity and risk factors in a multi-ethnic population. AB - BACKGROUND: There has been a relative lack of epidemiological data on generalised anxiety disorder (GAD) in Southeast Asia. A previous study reported a lifetime prevalence of 1.5% and highlighted low preference for seeking professional help and consultation by persons suspected to be suffering from mental health problems. The present study is part of a National Mental Health survey of adults conducted from February 2003-March 2004 specifically assessing anxiety and depression in Singapore. In this paper we report on prevalence, co-morbidity and risk factors associated with GAD. METHODS: We interviewed 2,847 households from an ethnically stratified random sample of adults aged 20-59 years who were Singapore citizens or permanent residents. The General Health Questionnaire and Schedule for Clinical Assessment of Neuropsychiatry were administered, which generated Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV) diagnoses of GAD. We assessed socio-demographic correlates, life events, medical and other psychiatric co-morbidities related to GAD. RESULTS: Lifetime prevalence of GAD was 3.3%, current prevalence is 3.0%. Female to male ratio is 3.6:1. GAD was significantly associated (p<0.001) with the presence of other psychiatric co-morbidities, including major depressive disorder, dysthymia, panic disorder, agoraphobia and social phobia. Prevalence increased in older individuals, with the odds of association greatest in subjects with three or more co-morbid medical conditions [adjusted odds ratio (OR) 3.66]. Those who had experienced one or more threatening life events showed increased odds of association with GAD. Chinese ethnicity, the divorced and persons from both the upper and the lowest socio-economic status had highest odds of association with GAD. CONCLUSIONS: We challenge established notions that GAD tends to be a disorder of the socially disadvantaged. Life events are important as precipitating factors in GAD, and uniquely different types of events appear to affect both extremes of social classes. High co-morbidity associations with current GAD are grounds for concern. This may suggest failure to seek treatment, hence giving rise to an increase in severity of the primary condition. PMID- 16249973 TI - Autoantibodies in children with chronic hepatitis B infection and the influence of interferon alpha. AB - BACKGROUND/AIMS: One of the serious side effects of interferon-a (IFN) is the possible induction of autoimmunity. However, data concerning children with chronic hepatitis B (HBV) infection is limited with conflicting results. The aim of this study was to evaluate the frequency of autoantibody positivity in children with chronic HBV infection and to assess whether IFN treatment has any influence on exacerbation of serological or clinical parameters of autoimmunity. METHODS: 61 children (32 female, mean age 7.5+/-3.8 years) were evaluated in two groups. Group I (29 patients) received 5 x 106 U/m2 IFN-a and group II (32 patients) 10 x 106 U/m2 IFN-a three times per week for six months. Autoantibody levels (anti-TPO, anti-Tg, AMA, ASMA, LKM-1, ANA, ds-DNA) and Ig G, A and M were analyzed before and after IFN treatment and 12 months after completion of therapy. RESULTS: No significant difference in autoimmune antibody positivity rate was observed between the two groups when compared at the beginning of the study and at the end of IFN treatment separately. SMA positivity rate was shown to significantly increase in group I after treatment was completed (p<0.05). None of the patients positive for autoantibodies showed further laboratory or clinical signs of autoimmunity. Thyroid hormones were within normal range in patients positive for anti-thyroid antibodies; however, thyrotropin-releasing hormone (TRH) stimulation test revealed subclinical hypothyroidism. All antibodies disappeared 12 months after completion of therapy. Overall, autoantibody positivity, pre- and posttreatment, were 16.3% and 54%, respectively (p<0.05). Age, sex, hepatitis activity index (HAI) score, HBV load and the dose of IFN had no influence on autoantibody formation. Complete and sustained response rates were similar in children with and without autoantibody. CONCLUSIONS: Autoantibody formation may occur in children with chronic HBV infection. IFN treatment leads to significant autoantibody formation, but this causes no organ dysfunction except for antithyroid antibodies associated with subclinical hypothyroidism. These results suggest that neither the presence of autoantibodies in choronic hepatitis B nor their development during IFN therapy is associated with severe autoimmune disorders in children with chronic HBV infection. PMID- 16249974 TI - Recent success of pantoprazole -or lansoprazole- based clarithromycin plus amoxicillin treatment in the eradication of Helicobacter pylori. AB - BACKGROUND/AIMS: There are some reports showing that resistance of Helicobacter pylori (H. pylori) to clarithromycin has increased in recent years. We aimed to investigate the current success of a most popular first-line eradication regimen by using two different proton pump inhibitors: lansoprazole and pantoprazole. METHODS: Ninety patients with H. pylori-positive functional dyspepsia were randomized to receive pantoprazole 40 mg b.i.d. or lansoprazole 30 mg b.i.d. in addition to amoxicillin 1,000 mg and clarithromycin 500 mg twice daily for 14 days in a multicenter study. H. pylori infection was determined by histological examination and a rapid urease test. A follow-up endoscopy was performed to assess the H. pylori eradication six weeks after the end of therapy. RESULTS: Seventy-nine patients completed the study protocol properly. The H. pylori eradication rates according to per protocol analysis were 70% in group pantoprazole, amoxicillin and clarithromycin (28/40) and 69.2% in group pantoprazole, amoxicillin and clarithromycin (27/39). The eradication rates according to intention to treat analysis were 62.2% and 60% in lansoprazole, amoxicillin, clarithromycin, pantoprazole, amoxicillin, clarithromycin groups, respectively. The eradication rates were similar in both protocols (p>0.05). CONCLUSIONS: The most popular first-line eradication protocols of H. pylori achieved only a moderate success in the current study. Alternative therapy options are needed instead of clarithromycin-based triple treatment for eradication of H. pylori. The choice of proton pump inhibitor is not important in the eradication rate of H. pylori. PMID- 16249975 TI - The effect of rabeprazole alone or in combination with H2 receptor blocker on intragastric pH: a pilot study. AB - BACKGROUND/AIMS: Proton pump inhibitors have been widely used in recent years. However, there are studies suggesting that proton pump inhibitors may not control the gastric acidity effectively during the night, especially in gastroesophageal reflux disease. It has therefore been suggested that H2 receptor blockers should be added to the therapy. The aim of our study was to evaluate the effects of proton pump inhibitors alone or in combination with H2 receptor blockers on gastric acidity with 24-hour gastric pH monitoring. METHODS: Esophagogastroscopy and 24-hour gastric pH monitoring were performed on 10 patients with dyspeptic symptoms. No patient had antacidity. All patients had erosive antral gastritis. Patients were randomized to two groups as either proton pump inhibitor therapy group (rabeprazole 20 mg/day, p.o.) or proton pump inhibitor + H2 receptor blocker therapy group (rabeprazole 20 mg/day, p.o. + famotidine 40 mg/day, p.o). After one month of treatment, 24-hour gastric pH monitoring was re-performed. RESULTS: Seven female and three male patients were enrolled into the study. The mean age was 51.1+/-11.56 years. All patients had antral erosive gastritis. Gastric pH was measured as less than 4 in 81.4% of the 24-hour period prior to rabeprazole treatment. With rabeprazole treatment this ratio decreased to 27.6% (p<0.05). These ratios were 86.3% and 4.55%, respectively, in the group that received combination therapy (p<0.05). CONCLUSIONS: Combination therapy with H2 receptor blockers and proton pump inhibitors seemed to control intra-gastric pH better than proton pump inhibitors alone. Use of H2 receptor blockers and proton pump inhibitors in combination to control intra-gastric pH is more beneficial. PMID- 16249976 TI - Hydatid acute pancreatitis. AB - BACKGROUND/AIMS: Hydatid acute pancreatitis is a rare condition, mostly reported as case presentations. METHODS: A series of eight patients with hydatid acute pancreatitis, referred between January 1990 and January 2003, are reported. All patients presented acute pancreatitis confirmed with clinical presentation, radiologic examination and laboratory findings. All patients had elevated levels of blood amylase value (more than 500 U/L). Five patients (62%) had high bilirubin levels (2.1 to 3.4 mg/dl) during the initial hospitalization. Computed tomography findings revealed acute pancreatitis in four patients; two had associated pseudocyst formation. RESULTS: Endoscopic retrograde cholangiopancreatography was performed on all patients and revealed hydatid cystic material in the common bile duct secondary to cystobiliary rupture in all patients. All patients underwent endoscopic sphincterotomy that was performed after dilatation with extractor balloon, and hydatid material was removed in all. Six patients were operated on after the initial episode subsided. Drainage of the cyst, appropriate cavity management and T-tube drainage of the common bile duct was employed in all patients to control bile leakage after the operation. Scolices and hydatid membrane were detected during common bile duct exploration in all patients due to presentation of cystobiliary rupture. There was no mortality. Postoperative pulmonary infection and wound infection were encountered in one patient each. During two to 13 years' follow-up, one patient developed recurrent hydatid disease. Recurrent pancreatitis did not occur. CONCLUSIONS: Hydatid acute pancreatitis is a rare condition. However, it should be remembered in patients with abdominal pain, especially in endemic areas. PMID- 16249977 TI - Value of endoscopic ultrasonography for upper gastrointestinal stromal tumors: a single center experience. AB - BACKGROUND/AIMS: Gastrointestinal stromal tumors are the rarely seen tumors of the gastrointestinal tract. The aim of the present study was to review the patients diagnosed as upper gastrointestinal stromal tumor by endoscopic ultrasonography. METHODS: Twenty-five patients diagnosed as upper gastrointestinal stromal tumor, between 1999 and 2004, were reviewed retrospectively. RESULTS: The reason for performing upper gastrointestinal system endoscopy was nonspecific upper gastrointestinal system symptoms in most (76%) of the patients. The other causes were upper gastrointestinal bleeding and dysphagia in 16% and 8% of the cases, respectively. Lesions were located in the stomach in 17 (68%), in the esophagus in six (24%), and in the duodenum in two (8%) patients. Endoscopic ultrasonographic evaluation revealed that all of the lesions arose from the muscularis propria. In 18 (72%) patients, tumors were less than 3 cm in diameter, homogeneous and hypoechoic in appearance with regular borders, concordant with benign tumor. In five (20%) patients, lesions had heterogeneous echoic appearance with anechoic spaces, two of which were larger than 3 cm and also showed irregular borders, suggesting malignancy. Surgical therapy was performed in five (20%) patients because of upper gastrointestinal bleeding or suspicion of malignancy by endoscopic ultrasonographic evaluation. Histopathological examination confirmed the diagnosis in all these patients. CONCLUSIONS: Endoscopic ultrasonographic evaluation is very useful in diagnosis and for choosing the therapeutic method for patients with upper gastrointestinal stromal tumor. PMID- 16249978 TI - Antineutrophil cytoplasmic autoantibodies and anti-Saccharomyces cerevisiae antibodies in inflammatory bowel diseases. AB - BACKGROUND/AIMS: Perinuclear antineutrophil cytoplasmic autoantibody is a marker for ulcerative colitis, and anti-Saccharomyces cerevisiae antibody is known to be associated with Crohn's disease. The purpose of this study was to search the value of detecting perinuclear antineutrophil cytoplasmic autoantibody and anti Saccharomyces cerevisiae antibody for the diagnosis of Turkish inflammatory bowel disease patients. METHODS: Serum samples were obtained from 80 patients with ulcerative colitis, 61 patients with Crohn's disease and 40 healthy controls. Determination of both anti-Saccharomyces cerevisiae antibody and antineutrophil cytoplasmic autoantibody was performed with the standardized enzyme-linked immunosorbent assay. RESULTS: In cases with ulcerative colitis, 65% tested seropositive for antineutrophil cytoplasmic autoantibody, whereas the controls showed 2.5% positivity. In cases with Crohn's disease, 63.9% tested seropositive for anti-Saccharomyces cerevisiae antibody, whereas the controls showed 2.5% seropositivity. The combination of a positive anti-Saccharomyces cerevisiae antibody test and a negative antineutrophil cytoplasmic autoantibody yielded a sensitivity and specificity of 32.0% and 97.5%, respectively. The combination of a positive perinuclear antineutrophil cytoplasmic autoantibody and a negative anti-Saccharomyces cerevisiae antibody test yielded a sensitivity and specificity of 44.2% and 97.5%, respectively. CONCLUSIONS: Both serologic tests may aid in the differential diagnosis of inflammatory bowel disease. PMID- 16249979 TI - The comparative effects of calcium channel blockers in an experimental colitis model in rats. AB - BACKGROUND/AIMS: In this study two calcium channel blockers (CCB), diltiazem and verapamil, which demonstrate their effects on two different receptor blockage mechanisms, were assessed comparatively in an experimental colitis model regarding the local and systemic effect spectrum. METHODS: Eighty male Swiss albino rats were divided into eight groups (n:10 each): Group I) colitis was induced with 1 ml 4% acetic acid without any medication. Group II) Sham group. Group III) Intra-muscular (IM) diltiazem was administered daily for five days before inducing colitis. Group IV) IM verapamil was administered daily for five days before inducing colitis. Group V) Transrectal (TR) diltiazem was administered with enema daily for two days before inducing colitis. Group VI) TR saline was administered four hours before inducing colitis. Group VII) TR diltiazem was administered with enema four hours before inducing colitis. Group VIII) TR verapamil was administered with enema four hours before inducing colitis. All subjects were sacrificed 48 hours after the colitis induction. The distal colon segment was assessed macroscopically and microscopically for the grade of damage, and myeloperoxidase (MPO) activity was measured. RESULTS: All the data of the control colitis group (group I), including the microscopic, macroscopic and MPO activity measurements, were significantly higher than in the groups in which verapamil and diltiazem were administered over seven days (3.100+/-0.7379 to 1.300+/-0.9487 and 1.600+/-0.9661) (p<0.05). The data of the Sham group, group II, were less than the other groups in which colitis was induced (p<0.05). For the local effect spectrum, after the assessment of groups V VIII, the control colitis group (group I) and group VI had significantly higher values than the others (3.300+/-0.4830 to 1.800+/-0.6325 and 1.700+/-0.8233 (p<0.05). CONCLUSIONS: Calcium channel blockage has systemic and local effects on the colitis model. PMID- 16249980 TI - Factor V Leiden, prothrombin G20210A and MTHFR gene mutations in inflammatory bowel disease. AB - BACKGROUND/AIMS: Thromboembolic events are more common in patients with inflammatory bowel disease than in the normal population; however, the reason for the increased prevalence is not clear. The aim of this study was to evaluate the prevalence of factor V Leiden, prothrombin G20210A and methylene tetrahydrofolate reductase (MTHFR) gene mutations in IBD patients followed in our outpatient clinic. METHODS: Thirty-four patients with ulcerative colitis and 28 patients with Crohn's disease and 80 healthy controls were included in the study. No patient had a history of previous thromboembolism. Factor V Leiden, prothrombin G20210A and MTHFR gene mutations were studied. RESULTS: Heterozygote factor V Leiden mutation was found in five (6.25%) control patients and in two (3.2%) IBD patients. Heterozygote MTHFR mutation was obtained in seven (11.3%) IBD patients and in five (6.25%) controls. Heterozygote prothrombin G20210A mutation was found in two (2.5%) and homozygote MTHFR mutation in one (1.25%) control patient. There was no statistical difference between the IBD group and healthy controls. CONCLUSIONS: Genetic mutations that could increase the thrombosis risk were not found to be different in IBD versus the normal population in our study. PMID- 16249982 TI - Acustimulation of the Neiguan point during gastroscopy: its effects on nausea and retching. AB - BACKGROUND/AIMS: Gastroscopic procedure causes nausea and retching in many patients. Recently, alternative methods have been employed in an effort to reduce these symptoms. The Neiguan point (P6) is an acupuncture point that has been used for approximately 3,000 years to overcome gastric symptoms including nausea and retching. The aim of this study was to investigate the effects of the stimulation of the P6 acupoint on swallowing, nausea and retching during the gastroscopic procedure. METHODS: Three hundred and twenty-seven patients who visited the gastroenterology unit for dyspeptic complaints were included in the study. A portable transcutaneous electrical nerve stimulation device (Reliefband; Maven Lab, Yuba City, CA) was used for acustimulation. The device was attached 15 minutes before the endoscopic procedure and no sedation was applied. The device was turned on in 78 patients (Group 1). The device was attached but not turned on in another 79 patients (Group 2). In Group 3, the device was attached to the Sham point (n: 79). In Group 4 the procedure was performed with no attachments (n: 77). Fourteen patients dropped out of the study because esophagogastroduodenoscopy could not be completed due to patient intolerance or to obstruction in the upper gastrointestinal tract. After the procedure, each patient's opinion about the severity of nausea and retching was measured on a visual analogue scale. Distress in swallowing and the impression of the endoscopist during the procedure were scored from 1 to 4. Patients were queried regarding their willingness to undergo re-endoscopy. RESULTS: Groups were compared regarding their distress in swallowing the endoscope, nausea and retching, the impression of the endoscopist during the procedure and their acceptance of re-endoscopy. Groups 1, 2, 3 and 4 were compared using the chi square test, and no significant difference was observed between the groups (p>0.05). CONCLUSIONS: Acustimulation of the Neiguan (P6) acupoint does not relieve patients of the nausea observed during gastroscopy, and its application does not facilitate the procedure. PMID- 16249981 TI - Acute gastrointestinal bleeding due to primary aortoduodenal fistula: report of two rare cases. AB - Primary aortoduodenal fistulas are among the rare causes of gastrointestinal hemorrhage and are defined as communications between the native abdominal aorta and the duodenum. The mortality rate is very high if undiagnosed and untreated. Two male patients, 61- and 76-years-old, were admitted to the emergency unit at different times with the chief complaints of abdominal pain, gastrointestinal hemorrhage and pulsatile mass in their abdomen. The first case experienced sudden massive upper gastrointestinal bleeding while being prepared for an emergency operation in the intensive care unit, and cardiac arrest developed within a few minutes. After resuscitation and successful surgical operation, the patient woke up without any neurological defect or sequelae and was extubated at the 9th postoperative hour. The second patient, who had been wounded by gun shot 30 years previously was admitted to the hospital because of simple gastrointestinal hemorrhage. A para-aortic pseudo-aneurysm connected with the duodenum was diagnosed by computed tomography. After successful surgical operation, the patient was discharged. In this report, a case of ruptured primary aortic aneurysm and another case of para-aortic pseudo-aneurysm connected with the duodenum, both of which were treated successfully by surgical operation, are presented. PMID- 16249983 TI - Ulcerative colitis case beginning during pregnancy in a patient with antiphospholipid antibody syndrome. AB - It is well known that there may be an increased tendency for thrombosis in inflammatory bowel disease (IBD). This study presents a case with antiphospholipid antibody syndrome with ulcerative colitis diagnosed during pregnancy. A 30-year-old female patient, in her 6th week of pregnancy, applied to our clinic with complaints of abdominal pain and bloody diarrhea. She had a history of three abortions. She had been given aspirin and heparin treatment due to high levels of anticardiolipin antibody (ACA) in previous examinations that led to the diagnosis of antiphospholipid antibody syndrome. As the left colonoscopic examination of the patient showed ulcerative colitis, aspirin treatment was replaced with mesalazine (750 mg/day, in 3 equal doses). There was a significant relief in complaints of the patient. Medical treatment was interrupted three days before delivery. The patient delivered a baby, at full term, of 3.6 kg by cesarean section. No adverse effects were observed in the mother or the baby due to the medical treatment. After delivery, mesalazine treatment was restarted. This case is interesting due to the co-diagnosis of IBD in a pregnant woman with antiphospholipid antibody syndrome. Successful and uncomplicated treatment of the patient with mesalazine should also be noted. PMID- 16249984 TI - A rare case of rectal prolapse associated with rectal adenocarcinoma: case report. AB - Despite the fact that colorectal polyps and solitary rectal ulcers may be present in conjunction with rectal prolapse, association between rectal prolapse and rectal cancer is very rare. As far as we could determine, there are only a few articles concerning rectal cancer in association with rectal prolapse in the literature. This case, a 63-year-old female patient, had suffered from a rectal prolapse since childhood and presented as a case of rectal cancer. At presentation, she complained of constipation and rectal bleeding for the previous six months. At physical examination there was a relaxed anal sphincter and a large reddish mass protruding via the anal canal when the patient strained. There was a fungating lesion in the upper left part of the rectal mucosa. An incisional biopsy was performed, the histopathological result of which was adenocarcinoma of the rectum. Changes in bowel habits, chronic constipation and chronic irritation seen in rectal prolapse may be responsible for the development of rectal cancer. Thus, a detailed history, digital rectal examination and rectosigmoidoscopic examination are important, particularly in patients with long-term rectal prolapse. PMID- 16249985 TI - Adenocarcinoma of the ileocecal valve: report of a case. AB - Tumors of the ileocecal valve are rarely mentioned, and are mostly defined radiologically. In this manuscript, we report a case of ileocecal valve adenocarcinoma diagnosed endoscopically. The case is a male patient, 78-years old, with abdominal pain and weight loss in whom a mass was palpated in the lower right quadrant. Abdominal ultrasound examination followed by colonoscopy and pathological diagnosis revealed adenocarcinoma of the ileocecal valve confirmed by surgical diagnosis. We report this case because of its rarity and because there is no endoscopic image in the medical literature. PMID- 16249986 TI - A giant villous adenoma case mimicking right colon carcinoma. AB - An 85-year-old woman was admitted to our hospital due to pain and swelling in her right inguinal region. She had a right inguinal hernia. Abdominal computerized tomography revealed an 8 x 8 cm cecal mass and also a 13.5 cm segmental asymmetric nodular thickening of the cecum. Colonoscopic examination revealed a sessile polypoid mucus-secreting mass mimicking carcinoma that narrowed the cecal lumen. Histopathological examinations of sections from colonoscopic biopsy materials on light microscopy revealed villous adenoma morphology. The patient underwent operation, and an 18 x 6 x 4 cm mass, which partially obstructed the cecum, was seen. A right hemicolectomy was performed for complete excision. Histopathological examination revealed a pure villous adenoma, and there was no sign of malignant degeneration. On the 6th postoperative day, the patient was discharged from the hospital. During follow-ups at three-month intervals throughout one year, no abnormal colonoscopic or laboratory findings were assessed. We believe that, in older patients with inguinal hernia, presence of intraabdominal mass should be considered. Furthermore, we showed in this report that villous adenomas can reach significant dimensions without causing any obstructing signs or electrolyte imbalance and can mimic colon carcinoma. PMID- 16249989 TI - Redox-switchable phase tags for recycling of homogeneous catalysts. PMID- 16249990 TI - The diverse world of silicon chemistry: 14th International Symposium on Organosilicon Chemistry (ISOS XIV) in conjunction with the 3rd European Organosilicon Day, Wurzburg, Germany, 31st July to 5th August, 2005. PMID- 16249991 TI - The impact of bacterial genomics on natural product research. AB - "There's life in the old dog yet!" This adage also holds true for natural product research. After the era of natural products was declared to be over, because of the introduction of combinatorial synthesis techniques, natural product research has taken a surprising turn back towards a major field of pharmaceutical research. Current challenges, such as emerging multidrug-resistant bacteria, might be overcome by developments which combine genomic knowledge with applied biology and chemistry to identify, produce, and alter the structure of new lead compounds. Significant biological activity is reported much less frequently for synthetic compounds, a fact reflected in the large proportion of natural products and their derivatives in clinical use. This Review describes the impact of microbial genomics on natural products research, in particularly the search for new lead structures and their optimization. The limitations of this research are also discussed, thus allowing a look into future developments. PMID- 16249992 TI - Nanosized [Pd52(CO)36(PEt3)14] and [Pd66(CO)45(PEt3)16] clusters based on a hypothetical Pd38 vertex-truncated nu3 octahedron. PMID- 16249993 TI - Two-component graded deposition of biomolecules with a double-barreled nanopipette. PMID- 16249994 TI - Evidence that interaction between neuregulin 1 and its receptor erbB4 increases susceptibility to schizophrenia. AB - There is now strong evidence that Neuregulin 1 (NRG1) is a susceptibility gene for schizophrenia. NRG1 mediates some of its effects through the tyrosine kinase receptor erbB4, and analysis of gene knock-out animals suggests that the functional interaction of NRG1 and erbB4 mediates behaviors that may model some aspects of the schizophrenia phenotype in mice. Given these findings, we have sought evidence for association between schizophrenia and erbB4. Mutation screening of erbB4 in 14 DSMIV schizophrenics revealed 15 SNPs, none of which were nonsynonymous. Analysis of the allele frequencies of each SNP in pools of 368 DSMIV schizophrenics and 368 controls provided modest evidence for association with two of the SNPs, although individual genotyping in an extended sample of 680 cases did not confirm this. However, we did find evidence for a significant interaction between the NRG1 "Icelandic" schizophrenia risk haplotype and erbB4 (P = 0.019). The NRG1 and erbB4 interacting marker was further genotyped in an independent sample of 290 cases and 634 controls from Dublin. Interaction between NRG1 and erbB4 remained significant in the combined sample of 970 cases and 1,341 controls, OR = 2.98 (CI: 1.16-7.64), P = 0.01, although it only showed a trend in the Dublin sample alone (P = 0.11, two tailed). Our data require independent replication, but tentatively suggest that NRG1 may mediate its effects on schizophrenia susceptibility through functional interaction with erbB4, and that genetic interaction between variants at the two loci increases susceptibility to schizophrenia. PMID- 16249995 TI - Association of a functional serotonin transporter gene polymorphism with binge eating disorder. AB - The pathophysiological mechanisms underlying binge eating disorder (BED) are poorly understood. There is evidence that abnormalities in brain serotonin (5HT) play an important role in binge eating behavior, therefore genes involved in 5HT transmission, such as the 5HT transporter (5HTT) gene, may contribute to the biological vulnerability to BED. We examined whether the polymorphism of the promoter of the 5HTT gene, consisting of a long (L) and a short (S) variant, was associated with BED. Seventy-seven obese or non-obese women with BED, and 61 normal weight control women were genotyped at the 5HTT gene linked polymorphism (5HTTLPR). Statistical analysis showed that both the LL genotype and the L allele of the 5HTTLPR were significantly more frequent in BED subjects. Moreover, the L allele was associated with a moderate but significant risk to develop BED (OR = 2.01, CI = 1.33-3.57). Although these data should be regarded as preliminary because of the small size of our sample, they suggest that the 5HTTPRL may contribute to the genetic susceptibility to BED. PMID- 16249996 TI - Effects of feed restriction on fertility in female rats. AB - BACKGROUND: Feed restriction with its resultant body weight loss impacts the rodent estrous cycle; however, the manifestation of these changes in a regulatory study design has not been documented. This study reports the effects of feed restriction in the context of an FDA regulatory submission. METHODS: Adult female rats (n = 20/group; weighing approximately 200 g each) were provided rodent chow ad lib (control) or at 20, 15, 10, or 7.5 g/rat/day (g/day) during a 2-week pre mating phase, throughout the mating phase, and up to gestation day (GD) 7. On GD 8, all animals were provided ad lib feed until necropsy on GD 14. Estrous cyclicity, mating, and fertility parameters were evaluated. RESULTS: Ad lib rats consumed approximately 20 and 28 g/day during the pre-mating and gestation phases, respectively. All measured fertility parameters in the 20 g/day group were similar to control values. In the 15 g/day group, body weight was reduced by 16% at 2 weeks, prolonged diestrus occurred, and fertility was compromised due to reductions in corpora lutea. Within 2 weeks, mean body weight in groups receiving < or = 10 g/day was reduced by > or = 29% compared to ad lib values, and overt changes in estrous cyclicity, mating, and fertility occurred. The 7.5 g/day group was not sustainable beyond the pre-mating phase. CONCLUSIONS: For this study type, feed intake at < or = 50% ad lib values (< or = 10 g/day) was inadequate due to the magnitude and rapidity of body weight effects. Estrous parameters appeared slightly more sensitive than functional measures, as body weight changes of approximately 16% appeared near the threshold of changing routinely calculated estrous cycle parameters and were later associated with reduced fertility. In general, body weight differences of 10-15% by themselves were not adverse to normal reproduction (20 g/day). PMID- 16249997 TI - Effects of prenatal perfluorooctane sulfonate (PFOS) exposure on lung maturation in the perinatal rat. AB - BACKGROUND: Perfluorooctane sulfonate (PFOS), found widely in wildlife and humans, is environmentally and metabolically stable. Environmental PFOS may be from its use as a surfactant, hydrolysis of perfluorooctanesulfonyl fluoride, and degradation of N-alkyl-perfluorooctanesulfonamide compounds formerly used in numerous applications. Prenatal exposure to PFOS in rodents causes neonatal mortality; treatment on gestation days (GD) 19-20 is sufficient to induce neonatal death in rats. Affected pups are born alive but present with labored breathing. Their lungs are pale and often do not expand fully on perfusion. METHODS: Pregnant Sprague-Dawley rats received 0, 25, or 50 mg/kg/day PFOS/K+ orally on GD 19-20. Lungs from GD 21 fetuses and neonates were prepared for histology and morphometry. Rescue experiments included co-administration of dexamethasone or retinyl palmitate with PFOS. Pulmonary surfactant was investigated with mass spectrometry in GD 21 amniotic fluid and neonatal lungs. Microarray analysis was carried out on PND 0 lungs. RESULTS: Histologically, alveolar walls were thicker in lungs of PFOS-exposed newborns compared to controls. The ratio of solid tissue:small airway was increased, suggesting immaturity. Rescue studies were ineffective. Phospholipid concentrations and molecular speciation were unaffected by PFOS. No changes in markers of alveolar differentiation were detected by microarray analysis. CONCLUSIONS: Morphometric changes in lungs of PFOS exposed neonates were suggestive of immaturity, but the failure of rescue agents and normal pulmonary surfactant profile indicate that the labored respiration and mortality observed in PFOS-treated neonates was not due to lung immaturity. PMID- 16249998 TI - Effects of feed restriction during organogenesis on embryo-fetal development in rabbit. AB - BACKGROUND: Appropriate maternal nutrition and body weight gain during pregnancy is well established as a major factor in healthy prenatal development in humans. Given the role of nutrition and body weight gain in normal development, pharmaceuticals intended to reduce appetite and promote weight loss will generate developmental toxicity data that may be challenging to interpret. To aid with this, the effects of feed restriction, and subsequent reduction in maternal body weight gain, on embryo-fetal development was investigated in the rabbit. METHODS: Groups of 15 pregnant New Zealand White rabbits were offered 150 (control), 110, 75, 55, 35, and 15 g feed/day from gestation day (GD) 7-19. Cesarean sections were carried out on GD 29 and fetuses were examined for external, visceral, and skeletal development. RESULTS: Maternal body weights at the end of the feed restriction period (GD 20) were 0.97, 0.98, 0.93, 0.94, and 0.86 x control for the 110, 75, 55, 35, and 15 g feed/day groups, respectively. Only at 15 g feed/day was there a net maternal body weight loss (the GD 20 body weight was 0.93 x the GD 6 body weight) at the end of the feed restriction period. Six does aborted in the 15 g feed/day group; there were no other abortions associated with feed restriction. Fetal body weight was significantly reduced at 75, 55, 35, and 15 g feed/day (0.95, 0.90, 0.86, and 0.84 x control, respectively). There were no external or visceral malformations or variations, and no skeletal malformations associated with feed restriction. The incidence of fetuses with sternebrae 5 or 6 unossified was increased at feed levels < or = 75 g/day. At a feed level of 35 g/day there was an increase in unossified metatarsals and metacarpals, and an increase in the number of fetuses with a reduced number of caudal vertebrae ossified. Although these findings were not increased at a feed level of 15 g/day, the lack of dose response was likely due to increased abortion and subsequent decrease in fetuses available for evaluation at 15 g feed/day. CONCLUSION: These data demonstrate that feed restriction to feed levels that produce substantial reductions in maternal body weight gain can result in developmental toxicity expressed by abortion, reduced fetal weight, and alterations in ossification. Abortion only occurred when feed was restricted to an amount that produced maternal body weight loss (15 g feed/day) whereas reduced fetal weight and increased incidence of fetuses with unossified sternebrae, metatarsals, metacarpals, or caudal vertebrae were noted at feed levels of < or = 75 g/day. There were no fetal malformations associated with feed restriction. PMID- 16250000 TI - SCN- binding to the charged lysines of histones end domains mimics acetylation and shows the major histone-DNA interactions involved in eu and heterochromatin stabilization. AB - SCN- binds to the charged amino group of lysines, inducing local changes in the electrostatic free energy of histones. We exploited this property to selectively perturb the histone-DNA interactions involved in the stabilization of eu and heterochromatin. Differential scanning calorimetry (DSC) was used as leading technique in combination with trypsin digestion that selectively cleaves the histone end domains. Euchromatin undergoes progressive destabilization with increasing KSCN concentration from 0 to 0.3 M. Trypsin digestion in the presence of 0.2 M KSCN show that the stability of the linker decreases as a consequence of the competitive binding of SCN- to the amino groups located in the C and N terminal domain of H1 and H3, respectively; likewise, the release of the N terminal domain of H4 induces an appreciable depression in both the temperature and enthalpy of melting of core particle DNA. Unfolding of heterochromatin requires, in addition to further cleavage of H4, extensive digestion of H2A and H2B, strongly suggesting that these histones stabilize the higher order structure by forming a protein network which extends throughout the heterochromatin domain. PMID- 16250002 TI - Crystal structure of the hypothetical protein TTHA1013 from Thermus thermophilus HB8. PMID- 16250001 TI - A promoter element of the CD-RAP gene is required for repression of gene expression in non-cartilage tissues in vitro and in vivo. AB - The cartilage-derived retinoic acid-sensitive protein (CD-RAP) gene is expressed predominately in cartilage. Previous studies in transgenic mice have shown that the DNA promoter segment from -2,251 bp to -2,068 bp of the CD-RAP gene contains elements critical for gene expression. Subsequent studies revealed both positive and negative regulatory motifs in this 183 bp element. Here we show that this element demonstrates activation or repression of gene expression in vitro and in vivo based on cell type and content of transcription factors. The distribution of Sox (positive) and C/EBP (negative) transcription factors in cell lines and in mouse tissues is consistent with their positive and negative roles. In transgenic mice, when the 183-bp element was removed from a 3,345-bp cartilage-specific CD RAP promoter, expression of the reporter gene became widespread, being observed in muscle, bone, lung, and liver in addition to cartilage. In vitro, mutation of the C/EBP site activated the inactive 3,345-bp CD-RAP gene promoter in myoblastic cells, suggesting that this site is responsible for (-2,079 bp) repression. These results indicate that the 183-bp element plays an important role in cartilage specific gene expression by acting as a chondrocyte-regulatory module repressing transcription in non-chondrocytes and contributing to activation in chondrocytes. This is the first report of a functional DNA element necessary for repression in non-cartilage tissues in vivo. PMID- 16250003 TI - Update of the molecular basis of familial hypercholesterolemia in The Netherlands. AB - Autosomal-dominant hypercholesterolemia (ADH) has been identified as a major risk factor for coronary vascular disease (CVD) and is associated with mutations in the low-density lipoprotein receptor (LDLR) and the apolipoprotein B (APOB) gene. Since 1991 DNA samples from clinically diagnosed ADH patients have been routinely analyzed for the presence of LDLR and APOB gene mutations. As of 2001, 1,641 index patients (164 index patients per year) had been identified, while from 2001 onward a more sensitive, high-throughput system was used, resulting in the identification of 1,177 new index patients (average=294 index patients per year). Of these 1,177 index cases, 131 different causative genetic variants in the LDLR gene and six different causative mutations in the APOB gene were new for the Dutch population. Of these 131 mutations, 83 LDLR and four APOB gene mutations had not been reported before. The inclusion of all 2,818 index cases into the national screening program for familial hypercholesterolemia (FH) resulted in the identification of 7,079 relatives who carried a mutation that causes ADH. Screening of the LDLR and APOB genes in clinically diagnosed FH patients resulted in approximately 77% of the patients being identified as carriers of a causative mutation. The population of patients with ADH was divided into three genetically distinct groups: carriers of an LDLR mutation (FH), carriers of an APOB mutation (FDB), and non-LDLR/non-APOB patients (FH3). No differences were found with regard to untreated cholesterol levels, response to therapy, and onset of CVD. However, all groups were at an increased risk for CVD. Therefore, to ultimately identify all individuals with ADH, the identification of new genes and mutations in the genes that cause ADH is of crucial importance for the ongoing national program to identify patients with ADH by genetic cascade screening. PMID- 16250004 TI - Autoinhibitory regulation of soluble adenylyl cyclase. AB - Soluble adenylyl cyclase is an evolutionarily conserved bicarbonate sensor that plays a crucial role in cAMP dependent processes that occur during mammalian fertilization. sAC protein is expressed at the highest levels in male germ cells, and is found to occur as one of two known isoforms: a truncated protein (sAC(t)) that consists almost exclusively of the two conserved catalytic domains (C1 and C2), and a full-length form (sAC(fl)) that contains an additional noncatalytic C terminal region. Several studies suggested sAC(t) was more active than sAC(fl). We now demonstrate that the specific activity of sAC(t) is at least 10-fold higher than the specific activity of sAC(fl). Using deletion analysis and a novel genetic screen to identify activating mutations, we uncovered an autoinhibitory region just C-terminal to the C2 domain. Kinetic analysis of purified recombinant sAC revealed this autoinhibitory domain functions to lower the enzyme's V(max) without altering its affinity for substrate or regulation by any of the known modulators of sAC activity. Our results identify an additional regulatory mechanism specific to the sAC(fl) isoform. PMID- 16250005 TI - Metabolism, protein content, and in vitro embryonic development of goat cumulus oocyte complexes matured with physiological concentrations of glucose and L lactate. AB - No information is available concerning how the maturation environment controls the metabolism of goat oocytes. The objectives of this experiment were to: (1) Determine the concentrations of glucose, lactate, and pyruvate in caprine follicular fluid; and (2) Investigate the effects of physiological concentrations of glucose and lactate in the in vitro maturation (IVM) medium on the metabolism (glycolysis and pyruvate oxidation), protein content, and developmental competence of caprine oocytes and cumulus-oocyte complexes (COCs). Abattoir derived COCs were matured for 18-20 hr in a defined, SOF-based medium containing 0.75, 1.5 (follicular fluid = 1.4 mM), or 3.0 mM glucose, and 3.0, 6.0 (follicular fluid = 7.1 mM), or 12.0 mM L-lactate. The protein content of oocytes and COCs was not affected (P > 0.05) by the concentration of glucose and lactate in the maturation medium. Increasing glucose and lactate decreased (P < or = 0.05) glycolytic activity of oocytes, without affecting (P > 0.05) pyruvate oxidation. In COCs, increasing glucose concentrations tended (P = 0.07) to decrease glycolysis. When metabolic activity was corrected for protein content (pmol/microg protein/3 hr), increasing glucose or lactate concentrations in the medium decreased (P < or = 0.05) pyruvate oxidation in oocytes, but increased (P < or = 0.05) pyruvate oxidation in COCs. Embryonic development (cleavage and blastocyst development, hatching, and cell number) was not affected (P > 0.05) by the glucose and lactate concentrations tested. These results indicate that concentrations of glucose and lactate in the medium have cell type-specific effects on metabolism of oocytes and COCs, but do not affect developmental competence within the range of concentrations tested. PMID- 16250006 TI - Murine Spam1 mRNA: involvement of AU-rich elements in the 3'UTR and antisense RNA in its tight post-transcriptional regulation in spermatids. AB - Sperm adhesion molecule1 (SPAM1), the best characterized hyaluronidase gene, is abundantly expressed in the testis. We attempted to overexpress mouse Spam1 via transgenesis using either the endogenous promoter in a BAC or a heterologous Protamine1 promoter for a Spam1 cDNA transgene. Although transgene-copy numbers ranged from 2 to 15 and transgenic transcripts were expressed, there was a general failure of overexpression of the RNA and protein in the testis of all seven founders. Also, three transgenic lines showed a modest downregulation or co suppression of the RNA for Spam1 and Hyal5, present on the BAC. We provide evidence for the potential involvement of two co-ordinating post-transcriptional regulatory processes in the failure of overexpression: abundant endogenous antisense RNA and adenosine-uridine (AU)-rich element-mediated regulation of RNA turnover. We demonstrate that AU-rich elements (AREs) in the 3'UTR of mRNAs, well known to interact with trans-acting proteins to target the RNA for (in)stability, are present in Spam1 RNA and specifically bind to six testicular cytoplasmic proteins. These AU-binding proteins (AUBPs) were virtually absent from the kidney where transcripts are rare, and were shown to interact with the cytoskeleton, which modulates mRNA turnover. In addition to a role in the RNAi pathway, antisense RNA can also modulate ARE-mediated regulation of mRNA by hybridizing to the AREs and specifically silencing their function. This potentially links the two processes in the regulation of Spam1 expression. We hypothesize that testicular Spam1 RNA is regulated post-transcriptionally by cis-acting ARE(s) in the 3'UTR which recognize AUBPs and which are modulated by antisense transcripts. PMID- 16250007 TI - Targeted suppression of E-cadherin gene expression in bovine preimplantation embryo by RNA interference technology using double-stranded RNA. AB - RNA interference (RNAi) has become acknowledged as an effective and useful tool to study gene function in diverse groups of cells. We aimed to suppress the expression of the E-cadherin gene during in vitro development of bovine preimplantation embryos using RNAi approach. In this experiment the effect of microinjection of E-cadherin and Oct-4 (as control) double-stranded (ds) RNA on the mRNA and protein expression level of the target E-cadherin gene was investigated. For this, a 496 bp long bovine E-cadherin and 341 bp long Oct-4 dsRNA sample were prepared using in vitro transcription. In vitro produced bovine zygotes were categorized into four treatment groups including those injected with E-cadherin dsRNA, Oct-4 dsRNA, RNase-free water, and uninjected controls. While the injection of E-cadherin dsRNA resulted in the reduction of E-cadherin mRNA and protein levels at the morula and blastocyst stage, the transcript and protein product remained unaffected in the Oct-4 dsRNA, water injected and uninjected control groups. The relative abundance of E-cadherin mRNA in the E-cadherin dsRNA injected morula stage embryos was reduced by 80% compared to the control group (P < 0.05). The Western blot analysis also showed a significant decrease in the E cadherin protein (119 kDa) in E-cadherin dsRNA injected embryos compared to the other three groups. Microinjection of E-cadherin dsRNA has resulted only 22% blastocyst rate compared to 38%-40% in water injected and uninjected controls. In conclusion, our results indicated the suppression of E-cadherin mRNA and protein has resulted in lower blastocyst rate and the RNAi technology is a promising approach to study the function of genes in early bovine embryogenesis. PMID- 16250008 TI - Factors affecting the efficiency and reversibility of roscovitine (ROS) block on the meiotic resumption of goat oocytes. AB - Goat oocytes from 2 to 4 and 0.8 to 1.2-mm follicles were freed (DOs) or not (COCs) of cumulus cells and cultured for different times in an inhibition medium supplemented with different concentrations of roscovitine (ROS). At the end of culture, oocytes were either cultured in a maturation medium for 24 hr and activated chemically for embryo development, or examined for GV chromatin configurations. Nuclear status was checked at different time points during maturation culture. Although both 200 and 250 microM ROS maintained 78-85% of oocytes at the GV stage for 24 hr, only oocytes blocked with 200 microM ROS developed to MII stage at a high rate after maturation culture. While few oocytes blocked with 200 microM ROS for 24 hr developed into morulae and none into blastocysts after activation, percentages of oocytes developing into morulae and blastocysts increased to the level of the control oocytes when the block time was reduced to 8 hr. While the GV and pMI stages were shortened with MI, and A/TI unaffected after oocytes were blocked for 8 hr, all the stages but A/TI were shortened after 24 hr of block. The sizes of nucleoli diminished with time and the GV chromatin configuration changed during ROS block. Significantly more DOs than COCs were blocked with 200 microM ROS, but none of the blocked DOs matured after drug withdrawal. However, maturation of the DOs improved significantly when ROS concentration was reduced to 150 microM or DOs were co-inhibited with COCs. The GV intact percentages of DOs did not differ after ROS inhibition with or without eCG, but those of COCs decreased significantly after ROS inhibited in the presence of eCG. When MII-incompetent oocytes from 0.8 to 1.2-mm follicles were inhibited with ROS for 8 and 24 hr prior to maturation culture, nuclear maturation improved significantly, activation rates were as high as that of the control oocytes, and some of the activated developed to 4- or 8-cell stages. It is concluded that (i) the efficiency and reversibility of ROS block was both drug concentration and exposure-time dependent; (ii) cumulus cells alleviated the toxicity of ROS on goat oocytes; (iii) eCG released goat oocytes from ROS block through the mediation of cumulus cells; (iv) ROS block quickened the nuclear maturation of goat oocytes and improved the developmental competence of meiosis incompetent oocytes, possibly due to a sustained nuclear activity during inhibition culture; (v) oocyte nuclear maturation and activation did not depend upon cumulus expansion, but the embryo development occurred in association with cumulus expansion. PMID- 16250009 TI - Hydrodynamic compression of young and adult rat osteoblast-like cells on titanium fiber mesh. AB - Living bone cells are responsive to mechanical loading. Consequently, numerous in vitro models have been developed to examine the application of loading to cells. However, not all systems are suitable for the fibrous and porous three dimensional materials, which are preferable for tissue repair purposes, or for the production of tissue engineering scaffolds. For three-dimensional applications, mechanical loading of cells with either fluid flow systems or hydrodynamic pressure systems has to be considered. Here, we aimed to evaluate the response of osteoblast-like cells to hydrodynamic compression, while growing in a three-dimensional titanium fiber mesh scaffolding material. For this purpose, a custom hydrodynamic compression chamber was built. Bone marrow cells were obtained from the femora of young (12-day-old) or old (1-year-old) rats, and precultured in the presence of dexamethasone and beta-glycerophosphate to achieve an osteoblast-like phenotype. Subsequently, cells were seeded onto the titanium mesh scaffolds, and subjected to hydrodynamic pressure, alternating between 0.3 to 5.0 MPa at 1 Hz, at 15-min intervals for a total of 60 min per day for up to 3 days. After pressurization, cell viability was checked. Afterward, DNA levels, alkaline phosphatase (ALP) activity, and extracellular calcium content were measured. Finally, all specimens were observed with scanning electron microscopy. Cell viability studies showed that the applied pressure was not harmful to the cells. Furthermore, we found that cells were able to detect the compression forces, because we did see evident effects on the cell numbers of the cells derived from old animals. However, there were no other changes in the cells under pressure. Finally, it was also noticeable that cells from old animals did not express ALP activity, but did show similar calcified extracellular matrix formation to the cells from young animals. In conclusion, the difference in DNA levels as reaction toward pressure, and the difference in ALP levels, suggest that the osteogenic properties of bone marrow-derived osteoblast-like cells are different with respect to the age of the donor. PMID- 16250011 TI - Expression of metalloproteinase-7 (matrilysin) in human blood and bronchoalveolar gamma/delta T-lymphocytes. Selective upregulation by the soluble non-peptidic mycobacterial phosphoantigen (isopentenyl pyrophosphate). AB - Human gammadelta T-lymphocytes are believed to regulate local immune defense and enhance resistance against invading microbes, although their precise function remains unknown. Herein, we addressed the question whether gammadelta T lymphocytes mediate these processes via synthesis of MMP-7, a protease closely associated with both epithelial repair and mucosal defense. Blood and bronchoalveolar gammadelta T-lymphocytes were cultured in the absence and presence of isopentenyl pyrophosphate (IPP) or TGF-beta1/IL-15 for 24 h, and assessed for the expression and synthesis of MMP-1, MMP-7, and MMP-9. Resting human gammadelta T-lymphocytes constitutively expressed MMP-9 mRNA, a marginal or no MMP-7 and MMP-1 mRNA. In the presence of IPP (3 microg/ml), expression of MMP 7 mRNA significantly increased, whereas TGF-beta1/IL-15 had no effect. Further, quiescent gammadelta T-lymphocytes obtained from bronchoalveolar lavage (BAL) fluid showed a weak or no MMP-7 mRNA signal which was raised significantly following stimulation with IPP. In Western blot analysis, a 28-kDa pro-matrilysin could be detected both in cell lysates (2 days) and supernatants (5 days) with a four- to sevenfold increased signal following IPP-stimulation of the gammadelta T lymphocytes. In conclusion, the data demonstrate for the first time that both human blood and BAL gammadelta T-lymphocytes express MMP-7 mRNA and synthesize MMP-7-protein. This unfolds a new perspective for the understanding of gammadelta T-lymphocyte function. PMID- 16250010 TI - Surface-attached PEO in the form of activated Pluronic with immobilized factor H reduces both coagulation and complement activation in a whole-blood model. AB - In the present work we have bound Pluronic, a class of triblock copolymers consisting of a block of polypropylene oxide (PPO) surrounded on each side by polyethylene oxide (PEO) blocks, to polystyrene surfaces and investigated the thrombogenicity and complement activation of this construct upon exposure to whole blood. The surface was highly inert towards coagulation, unfortunately at the expense of increased complement activation. We, therefore, as an alternative approach, used End-Group Activated Pluronic to conjugate factor H, a regulator of complement activation (RCA), to the surface. The bound factor H did not detach from the surface upon incubation with human serum. Furthermore, factor H bound in a physiological conformation could to a significant degree attenuate complement activation at the Pluronic surface. Thus, we have created a hybrid surface in which the coagulation-inert properties of the original Pluronic are supplemented with a specific complement-inhibitory effect. Medical device technology includes numerous potential applications for crosslinkers that are capable of specifically binding biomolecules to surfaces with retained activity. These applications include coupling of functional biomolecules to biomedical devices such as stents and grafts. The biomolecule may be an RCA, antibody, or other beneficial ligand. PMID- 16250012 TI - Tyrosine phosphatase SHP-2 regulates IL-1 signaling in fibroblasts through focal adhesions. AB - Interleukin-1beta (IL-1beta) mediates destruction of matrix collagens in diverse inflammatory diseases including arthritis, periodontitis, and pulmonary fibrosis by activating fibroblasts, cells that interact with matrix proteins through integrin-based adhesions. In vitro, IL-1beta signaling is modulated by focal adhesions, supramolecular protein complexes that are enriched with tyrosine kinases and phosphatases. We assessed the importance of tyrosine phosphatases in regulating cell-matrix interactions and IL-1beta signaling. In human gingival fibroblasts plated on fibronectin, IL-1beta enhanced the maturation of focal adhesions as defined by morphology and enrichment with paxillin and alpha actinin. IL-1beta also induced activation of ERK and recruitment of phospho-ERK to focal complexes/adhesions. Treatment with the potent tyrosine phosphatase inhibitor pervanadate, in the absence of IL-1beta, recapitulated many of these responses indicating the importance of tyrosine phosphatases. Immunoblotting of collagen bead-associated complexes revealed that the tyrosine phosphatase, SHP-2, was also enriched in focal complexes/adhesions. Depletion of SHP-2 by siRNA or by homologous recombination markedly altered IL-1beta-induced ERK activation and maturation of focal adhesions. IL-1beta-induced tyrosine phosphorylation of SHP-2 on residue Y542 promoted focal adhesion maturation. Association of Gab1 with SHP 2 in focal adhesions correlated temporally with activation of ERK and was abrogated in cells expressing mutant (Y542F) SHP-2. We conclude that IL-1beta mediated maturation of focal adhesions is dependent on tyrosine phosphorylation of SHP-2 at Y542, leading to recruitment of Gab1, a process that may influence the downstream activation of ERK. PMID- 16250014 TI - The adaptive immune response in neonatal cerebral white matter damage. AB - Hypotheses that inflammation contributes to neonatal cerebral white matter damage have evolved over the last three decades. The latest, expanded here, suggests that the adaptive immune system contributes to the intensity and duration of the processes that result in damage to cerebral white matter in the fetus and newborn. We propose several mechanisms by which fetal T lymphocytes could be activated during fetal exposure to infection. These include specific recognition of bacterial antigens, specific recognition of autoantigens, polyclonal activation by Toll-like receptors, and bystander activation by cytokines. PMID- 16250013 TI - Nectin-2 expression in testicular cells is controlled via the functional cooperation between transcription factors of the Sp1, CREB, and AP-1 families. AB - Nectin-2, a major protein component of the adherens junctions (AJs), is found between Sertoli cells and germ cells in the seminiferous epithelium. Recent studies have shown that the expression of nectin-2 gene in testis is crucial to maintain normal spermatogenesis since male knockout mice lacking nectin-2 gene are sterile and possess morphologically abnormal spermatozoa. However, the molecular mechanisms governing its basal transcription remain poorly understood. By the use of Sertoli and germ cell-lines (TM4 and GC-2spd(ts) cells, respectively) in transient transfection studies, we showed that the minimal mouse nectin-2 promoter was located between nucleotides -316 and -211 (relative to the translation start site). Two putative Sp1 motifs and one each of the CRE, AP1, and AP2 motifs were identified within this region. Mutational studies showed that these two Sp1 motifs cooperated synergistically with the CRE motif, but not the AP1 and AP2 motifs, to regulate nectin-2 gene transcription in both TM4 and GC 2spd(ts) cells. By EMSAs, we found that an AP-1 consensus sequence was able to inhibit DNA-protein complex formation with the CRE motif, suggesting an interaction between the AP-1 transcription factor (c-Jun) and CREB within the CRE motif. Overexpressions of CREB and c-Jun, but not c-Fos, also significantly increased the promoter activity, which suggests that CREB and c-Jun are the crucial transcription factors involved in regulating nectin-2 gene transcription. Chromatin immunoprecipitation assay has shown that, in vivo, CREB, c-Jun, and Sp1 family proteins are bound to the mouse nectin-2 promoter. Analysis of the staged tubules has confirmed that the cyclic expressions of CREB and nectin-2 coincide with the event of adherens junction restructuring between Sertoli cells and germ cells. The cross-talk between CREB, c-Jun, and Sp1 family protein is believed to be a major transcription machinery to drive nectin-2 expression in Sertoli cells. PMID- 16250015 TI - Normal and transforming functions of RUNX1: a perspective. AB - Converging studies from many investigators indicate that RUNX1 has a critical role in the correct maintenance of essential cellular functions during embryonic development and after birth. The discovery that this gene is also frequently mutated in human leukemia has increased the interest in the role that RUNX1 plays in both normal and transforming pathways. Here, we provide an overview of the many roles of RUNX1 in hematopoietic self-renewal and differentiation and summarize the information that is currently available on the many mechanisms of RUNX1 deregulation in human leukemia. PMID- 16250017 TI - Action fluency in Parkinson's disease: a follow-up study. AB - The impairment in action fluency task present in Parkinson's disease (PD) patients has been previously interpreted as an indicator of conversion from PD to PD with dementia or as a grammatical deficit for verbs and ascribed to a frontostriatal loop pathophysiology. In the present study, 20 patients with PD without dementia were longitudinally tested with overall cognitive decline scales and semantic, letter, and action fluency tasks in a 24-month follow-up study. In comparison with healthy age-matched controls, PD patients showed a stable and consistent impairment on action fluency without any sign of cognitive decline. Our findings suggest that action fluency task may be an early sign of impairment of frontostriatal circuits in PD and it cannot be considered an indicator of conversion from PD to PD with dementia. PMID- 16250016 TI - Expression of the mechanosensitive 2PK+ channel TREK-1 in human osteoblasts. AB - TREK-1 is a mechanosensitive member of the two-pore domain potassium channel family (2PK+) that is also sensitive to lipids, free fatty acids (including arachidonic acid), temperature, intracellular pH, and a range of clinically relevant compounds including volatile anaesthetics. TREK-1 is known to be expressed at high levels in excitable tissues, such as the nervous system, the heart and smooth muscle, where it is believed to play a prominent role in controlling resting cell membrane potential and electrical excitability. In this report, we use RT-PCR, Western blotting and immunohistochemistry to confirm that human derived osteoblasts and MG63 cells express TREK-1 mRNA and protein. In addition, we show gene expression of TREK2c and TRAAK channels. Furthermore, whole cell patch clamp electrophysiology demonstrates that these cells express a spontaneously active, outwardly rectifying potassium "background leak" current that shares many similarities to TREK-1. The outward current is largely insensitive to TEA and Ba2+, and is sensitive to application of lysophosphatidylcholine (LPC). In addition, blocking TREK-1 channel activity is shown to upregulate bone cell proliferation. It is concluded that human osteoblasts functionally express TREK-1 and that these channels contribute, at least in part, to the resting membrane potential of human osteoblast cells. We hypothesise a possible role for TREK-1 in mechanotransduction, leading to bone remodelling. PMID- 16250018 TI - Involvement of CSF-1 in generating a stroma-independent hematopoietic stem cell line. AB - The hematopoietic stem cell line, Myl-D7, is maintained by a self-renewing stem cell population that spontaneously generates myeloid, lymphoid, and erythroid progeny. MS-5 stromal cells are necessary for the growth of Myl-D7 cells. One component of the Myl-D7 cells proliferation activity released by MS-5 stromal cells was enriched by Q sepharose fractionation and shown to be colony stimulating factor-1 (CSF-1) by Western blotting, BAC1.2F5 cell bioassay and inhibition of Myl-D7 proliferation by CSF-1 antibody. The requirement of Myl-D7 cells for CSF-1 was also demonstrated independently by selecting for rare, stroma independent Myl-D7 mutant clones able to grow without stroma and additional factors. Eighty-nine stroma-independent mutant clones were obtained and belonged to two classes. The majority of mutants did not secrete any growth promoting activity. The second, rarer class of mutants releases a factor that stimulates proliferation/survival for up to several months and approximately half of the secretors express high levels of CSF-1 mRNA. Wild type Myl-D7 grown with supernatants from the secretor cells retained the stem cell phenotype. These data suggest that CSF-1 may act as a key factor in stroma-regulated hematopoiesis and cell-cell interaction. PMID- 16250019 TI - Movement disorders fellowship training program at Columbia University Medical Center in 2001-2002. AB - Data regarding the clinical experience of movement disorders subspecialty training are currently not available. In order to provide information to help design a well-rounded clinical experience, we reviewed the number and types of patients seen by all clinical movement disorders fellows over the course of 1 year at Columbia University Medical Center (CUMC). We conducted a retrospective analysis of all patients seen by four full-time clinical fellows and one part time fellow during the 2001-2002 academic year, using billing records to tabulate all clinic visits and patient charts to certify diagnosis. One thousand six hundred sixty-two patients (1,132 new and 530 follow-ups) were seen by five fellows. Each full-time fellow evaluated and treated a mean of 263 new patients, 116 follow-up patients, and 15 in-patient consultations. The part-time clinical fellow evaluated and treated 81 new patients and 65 follow-up patients. Approximately half of the new patients had idiopathic Parkinson's disease. Full time fellows saw equivalent numbers of patients but they evaluated few patients with uncommon movement disorders. Weekly video rounds allowed all fellows to see many more patients, aiding the recognition of movement phenomenology and enhancing understanding of diagnoses and treatment strategies. Thus, CUMC fellows evaluate a large number of patients with a wide range of diagnoses within a 1 year period. Video rounds allow greater exposure to uncommon diagnoses. Similar data from other movement disorder training programs are needed to help create guidelines for formal accreditation and to improve clinical training in this subspecialty. PMID- 16250020 TI - Acute reversible parkinsonism in Epstein-Barr virus-related encephalitis lethargica-like illness. AB - A 10-year-old boy presented with fever, headache, vomiting, and hypersomnolence. An akinetic-rigid syndrome with tremor, dysphagia, dysphonia, and sialorrhea, as well as pyramidal signs, developed. Slightly elevated protein content was found in the cerebrospinal fluid and serological investigations were suggestive of a primary Epstein-Barr virus infection. Magnetic resonance imaging (MRI) showed massive bilateral hyperintense striatal and punctiform periventricular lesions. After 2-month treatment with steroids and antiparkinsonian drugs, all features resolved without sequelae. Control MRI demonstrated only minimal residual lesions in both putamina. Strongly resembling the encephalitis lethargica-like syndrome, this case is an unusual presentation of parainfectious acute disseminated encephalitis. PMID- 16250021 TI - Multiple genetic and epigenetic interacting mechanisms contribute to clonally selection of drug-resistant tumors: current views and new therapeutic prospective. AB - Successful treatment of cancer requires a clear understanding of drug-resistance mechanism. Cancer patient are often treated with standard protocols without considering individual difference in chemosensitivity, whereas the efficacy of anticancer drug varies widely among individual patients. Since chemosensitivity involves multiple interacting factors, it is not sufficient to investigate a single gene or factor to fix chemoresistance. Along with affecting disease progression, the synergism between genetic and epigenetic abnormalities can contribute to convert a sensible tumor cell in a resistant one. Unlike genetic changes, epigenetic changes are potentially reversible. Therefore, treatment with DNA methylation inhibitors can reactivate the expression of genes improperly methylated and can reverse many aspect of cancer phenotype such as drug resistance. The demethylating agents are used in the treatment of several kind of tumor, but toxicity and the potential outcome on the normal methylation patterns have always been concern of researchers and clinicals. It is necessary to create individual chemosensitivity-chemoresistance maps in order to identify the combination of drugs for optimized treatments. An overview on genetic and epigenetic events contributing to clonally selection of chemotherapeutic resistant tumors is discussed. PMID- 16250022 TI - Association between restless legs syndrome and essential tremor. AB - After observing that several families with essential tremor (ET) clinically cosegregated with restless legs syndrome (RLS), we prospectively evaluated for the presence of RLS in 100 patients presenting to the Baylor College of Medicine with ET and prospectively examined all patients presenting with RLS for the presence of tremor during the same time frame. Of 100 consecutive ET patients (60 women and 75 with a family history of ET) seen over 19 weeks (current age, 65.2 +/- 16.3 years; age at tremor onset, 37.8 +/- 19.9 years) 33 met all criteria for RLS, of which 25 had never been diagnosed previously. A family history of RLS was reported in 57.6% of these 33 patients and was the only significant predictor of RLS in the ET population. Their International Restless Legs Syndrome Rating Scale score was 16.6 +/- 8.1. Over 19 weeks, we also examined 68 consecutive RLS patients (63.2% women and 54.4% with a family history of RLS) for the presence of tremor. Their current age was 55.8 +/- 14.4 years, and age at RLS onset was 33.7 +/- 19.5 years. Overtly pathological tremor was rare, but trace tremor was very common. Overall, we found a very high rate of undiagnosed RLS in patients presenting for tremor, but unlike other "secondary" forms of RLS, this finding was also associated with a high familial history of RLS, suggesting that they share some genetic similarities. PMID- 16250023 TI - [123I]-FP/CIT SPECT imaging for distinguishing drug-induced parkinsonism from Parkinson's disease. AB - Parkinsonism in patients taking neuroleptic medications might be induced by dopamine receptor blockade alone or by dopamine blockade with nigrostriatal dysfunction. The differentiation between Parkinson's disease (PD) and drug induced parkinsonism (DIP) is difficult to assess on clinical grounds alone. In this study, we have evaluated the clinical characteristics and striatal binding of (123)I-FP-CIT (N-omega-fluoropropyl-2beta-carboxymethoxy-3beta-{4 iodophenyl}tropane) in patients who developed DIP. A total of 20 patients (mean age, 62 +/- 13 years) who developed parkinsonism while on neuroleptic agents and 10 age-matched controls were enrolled. [123]-FP-CIT single-photon emission computed tomography (SPECT) was performed in all subjects. Neurological assessment was performed with the Motor part of the Unified Parkinson's Disease Rating Scale. [123]-FP-CIT binding of the entire striatum, caudate, and putamen was calculated. Patients were divided into two subgroups according to SPECT results for comparison of clinical characteristics. There were 9 patients who had normal scans and 11 who showed significantly diminished striatal binding, suggesting degeneration of the nigrostriatal system. Subanalyses of abnormal scans revealed significantly diminished binding in the caudate (P < 0.001 for right and left caudate) and putamen (P = 0.002 and P < 0.05 for right and left putamen, respectively). There were no differences in clinical features between patients with normal and abnormal scans. Symptoms included asymmetric tremor, bradykinesia, and rigidity in both groups. Freezing gait was present in two patients with normal scans. These results indicate that DIP is clinically indistinguishable from PD. Brain imaging with FP-CIT helps to determine whether DIP is entirely drug-induced or an exacerbation of subclinical PD. PMID- 16250024 TI - Catatonic stupor during the course of Parkinson's disease resolved with electroconvulsive therapy. PMID- 16250025 TI - Association of alpha-synuclein Rep1 polymorphism and Parkinson's disease: influence of Rep1 on age at onset. AB - The alpha-synuclein Rep1 polymorphism was studied in patients and controls in an ethnic Greek population. There was an association of allele 2 with risk of Parkinson's disease (PD; adjusted odd ratio = 3.25; 95% CI = 1.80-5.87). Survival analyses (Cox proportional hazards models) were employed to explore the influence of genotypes on age at onset of PD. Age at onset of carriers of at least one Rep1 allele 2 was earlier (3.6 years) compared to noncarriers (adjusted hazard ratio = 2.21; 95% CI = 1.58-3.10). Kaplan-Meier analysis also supported a dosage effect of Rep1 allele 2 on age at onset. For Rep1 allele 1, there was neither association with risk of PD nor influence on age at onset. This is the first study showing an influence of Rep1 polymorphism on age at onset of PD. PMID- 16250026 TI - Parkinsonism, dysautonomia, and intranuclear inclusions in a fragile X carrier: a clinical-pathological study. AB - A new tremor-ataxia syndrome, fragile X-associated tremor/ataxia syndrome (FXTAS), has been described among carriers of premutation expansions (55-200 CGG repeats) of the fragile X mental retardation 1 (FMR1) gene. The prevalence of FMR1 premutation alleles has been reported to be 1 in 813 among men. Patients with FXTAS may also have features of parkinsonism. Postmortem findings have been described in eight patients with FXTAS and detailed descriptions of the pathological features of this syndrome have been published in two of these. We present a detailed description of the postmortem findings in a third patient. The patient had parkinsonism and was a carrier of a premutation expansion in the FMR1 gene. As in previous reports, the most prominent finding was the presence of eosinophilic nuclear inclusions in neurons and astrocytes, loss of Purkinje cells, and regional vacuolation of the cerebral white matter. As in one previous report, nuclear inclusions were also present in ependymal and choroid plexus cells. A new finding is that of nuclear inclusions in both the adeno- and neurohypophysis. These findings confirm the diffuse nature of this pathology. Further studies of clinical-pathological correlation in a larger sample of brains would provide additional insight into the mechanisms of the tremor, ataxia, and parkinsonism in these patients. PMID- 16250027 TI - Betel quid: Its tremor-producing effects in residents of Araihazar, Bangladesh. AB - Betel quid consists of a betel nut (Areca catechu) and other additives. It is chewed throughout Asia and is the fourth most commonly used addictive substance in the world. Its stimulant properties are mediated by arecoline and other toxins. The neurological effects of betel quid use have not been studied extensively. To assess tremor, 100 residents of Araihazar, Bangladesh, were asked to draw spirals, which were rated by a blinded neurologist. The average tremor rating was 50% higher in exposed versus unexposed individuals. Betel quid use can produce clinically evident tremor. Population-based epidemiological studies of tremor in Asia should begin to take the effect of this stimulant into account. PMID- 16250028 TI - Corticomuscular coherence is increased in the small postural tremor of Parkinson's disease. AB - The mechanisms and electrophysiological characteristics of the postural tremor in Parkinson's disease (PD) have not been defined. We hypothesized that PD subjects with small amplitude postural tremor would show increased corticomuscular coherence at certain frequencies compared to PD subjects without visible tremor. Four groups of participants were studied: (1) Control without postural tremor, (2) Control with small amplitude postural tremor, (3) PD without postural tremor, and (4) PD with small amplitude postural tremor. Accelerometry and electroencephalography-electromyography fast-fourier transform and corticomuscular coherence spectra were generated. Findings showed (1) elevated corticomuscular coherence centered at 12-18 Hz in PD with small amplitude postural tremor; (2) 5-12 Hz accelerometer frequency peaks that did not shift with increasing weight loads in some individuals; and (3) 5-8 Hz accelerometer peaks that shifted frequency with increasing weight loads, consistent with a peripheral-mechanical oscillator in all groups. The small amplitude postural tremor in PD arises from heterogeneous oscillator mechanisms. The discovery of increased corticomuscular coupling shows cortical involvement in the small amplitude postural tremor of PD. PMID- 16250029 TI - Rhythmic tongue movements during sleep: a peculiar parasomnia in Costello syndrome. AB - We describe a peculiar parasomnia observed in four Costello infants, characterized by periodic rhythmic movements of the tongue. Ten Costello patients (4 male; age range 9 months to 29 years) underwent 1 full-night laboratory-based video polysomnography. The four youngest patients (2 male and 2 female; age range 9-31 months) presented during sleep repeated stereotyped movements of the tongue, producing a sucking-like or licking-like movement, mostly during non-rapid eye movement (NREM) sleep. Rhythmic tongue movements in Costello syndrome show the features of an NREM sleep parasomnia. Tongue movements during sleep probably originate from brainstem structures and could be facilitated by an impaired control of the oropharyngeal and tongue muscles. PMID- 16250030 TI - Parkinson's disease and LRRK2: frequency of a common mutation in U.S. movement disorder clinics. AB - The G2019S mutation in the LRRK2 gene is reportedly a common cause of familial Parkinson's disease (PD) and may also have a significant role in nonfamilial PD. The objective of this study was to assess mutation carrier frequency in PD patients from movement disorder clinics in the United States, stratified by family history, age at onset, and geography; to determine carrier frequency in a large and well-characterized control population; to examine segregation of mutation in families of patients; and to correlate genotype with clinical phenotype. One thousand four hundred twenty-five unrelated PD patients from movement disorder clinics in Oregon, Washington, and New York and 1,647 unrelated controls were studied. The G2019S mutation was detected using a TaqMan assay and verified by sequencing. Eighteen of 1,425 patients and one of 1,647 controls had the mutation. Carrier frequency (+/- 2SE) in patients was 0.013 +/- 0.006 overall, 0.030 +/- 0.019 in familial PD, 0.007 +/- 0.005 in nonfamilial PD, 0.016 +/- 0.013 in early-onset PD, and 0.012 +/- 0.007 in late-onset PD. Geographic differences were insignificant. Age at onset of mutation carriers ranged from 28 to 71 years. Mutation carriers were clinically indistinguishable from idiopathic PD. LRRK2 G2019S is the single most common pathogenic mutation linked to neurodegenerative disease to date. PMID- 16250031 TI - The CAR nuclear receptor and hepatocyte proliferation. PMID- 16250032 TI - Interferon alfa-2b in combination with ribavirin for the treatment of chronic hepatitis C in children: efficacy, safety, and pharmacokinetics. AB - Chronic hepatitis C virus (HCV) infection is usually asymptomatic in children, but significant liver disease may occur. We evaluated the efficacy, safety, and pharmacokinetics of interferon alfa-2b and ribavirin in children with chronic HCV. We determined the optimal ribavirin dose in an initial cohort of a phase 1 study and then subsequently used it, in combination with interferon alfa-2b, in a second cohort of this study and a phase 3 trial. The primary efficacy endpoint in all studies was sustained virological response, defined by undetectable serum HCV RNA 24 weeks after completion of therapy. All efficacy and safety analyses were performed on the intent-to-treat population. Children receiving interferon alfa 2b plus ribavirin 15 mg/kg/d in the phase 1 study had the maximum reduction in serum HCV RNA at treatment weeks 4 and 12 with an acceptable safety profile. This ribavirin dose was selected as optimal and used in all subsequent studies. In all, 46% (54/118) of optimally treated children achieved sustained virological response. Sustained virological response was significantly higher in children with HCV genotype 2/3 (84%) than in those with HCV genotype 1 (36%). Adverse events led to dose modification in 37 (31%) and discontinuation in 8 (7%). Multiple-dose interferon alfa-2b and ribavirin peak and trough concentrations and area-under-the-curve were similar between children and adults. In conclusion, interferon alfa-2b in combination with ribavirin is effective and safe in children with chronic hepatitis C virus. PMID- 16250033 TI - High prevalence of spontaneous portal-systemic shunts in persistent hepatic encephalopathy: a case-control study. AB - Large spontaneous portal-systemic shunts have been occasionally described in patients with cirrhosis. This study was undertaken to assess the prevalence of portal-systemic shunts in patients with cirrhosis with recurrent or persistent hepatic encephalopathy (HE) as compared with patients with cirrhosis without HE. Fourteen patients with cirrhosis with recurrent or persistent HE (cases) and 14 patients with cirrhosis without previous or present signs of overt HE matching for age and degree of liver failure (controls) were studied. Each patient underwent neurological assessment and cerebral magnetic resonance (MR) imaging to exclude organic neurological pathological conditions. HE evaluation included psychometric performance (Trail-Making Test A), electroencephalogram (EEG), mental status examination and grading, arterial, venous, and partial pressure of ammonia determination. The presence of portal-systemic shunts was assessed by portal venous phase multidetector-row spiral computed tomography (CT). Large spontaneous portal-systemic shunts were detected in 10 patients with HE and in only 2 patients without HE (71% vs. 14%; chi square = 9.16; df = 1.0; P = .002). The patients with HE presented ascites (P = .002) and medium/large esophageal varices (P = .02) less frequently than the control group. In conclusion, our study suggests that large spontaneous shunts may often sustain the chronicity of HE; the presence of large shunts should be sought in patients with cirrhosis with recurrent or persistent HE. PMID- 16250034 TI - Hepatocyte transplantation activates hepatic stellate cells with beneficial modulation of cell engraftment in the rat. AB - We investigated whether transplanted hepatocytes interact with hepatic stellate cells, as cell-cell interactions could modulate their engraftment in the liver. We transplanted Fischer 344 rat hepatocytes into syngeneic dipeptidyl peptidase IV-deficient rats. Activation of hepatic stellate cells was analyzed by changes in gene expression, including desmin and alpha-smooth muscle actin, matrix proteases and their inhibitors, growth factors, and other stellate cell associated genes with histological methods or polymerase chain reaction. Furthermore, the potential role of hepatic ischemia, Kupffer cells, and cytokine release in hepatic stellate cell activation was investigated. Hepatocyte transplantation activated desmin-positive hepatic stellate cells, as well as Kupffer cells, including in proximity with transplanted cells. Inhibition of Kupffer cells by gadolinium chloride, blockade of tumor necrosis factor alpha (TNF-alpha) activity with etanercept or attenuation of liver ischemia with nitroglycerin did not decrease this hepatic stellate cell perturbation. After cell transplantation, soluble signals capable of activating hepatic stellate cells were rapidly induced, along with early upregulated expression of matrix metalloproteinases-2, -3, -9, -13, -14, and their inhibitors. Moreover, prior depletion of activated hepatic stellate cells with gliotoxin decreased transplanted cell engraftment. In conclusion, cell transplantation activated hepatic stellate cells, which, in turn, contributed to transplanted cell engraftment in the liver. Manipulation of hepatic stellate cells might provide new strategies to improve liver repopulation after enhanced transplanted cell engraftment. PMID- 16250035 TI - Hepatic expression of ABC transporters G5 and G8 does not correlate with biliary cholesterol secretion in liver transplant patients. AB - The adenosine triphosphate (ATP)-binding cassette (ABC)-transporters ABCG5 and ABCG8 have been shown to mediate hepatic and intestinal excretion of cholesterol. In various (genetically modified) murine models, a strong relationship was found between hepatic expression of ABCG5/ABCG8 and biliary cholesterol content. Our study aimed to relate levels of hepatic expression of ABCG5 and ABCG8 to biliary excretion of cholesterol in man. From 24 patients who had received a liver transplant, bile samples were collected daily after transplantation over a 2-week period to determine biliary composition. Expression of ABCG5, ABCG8, MDR3, and BSEP was assessed by real-time polymerase chain reaction (PCR) in liver biopsy specimens collected before and after transplantation. Levels of hepatic ABCG5, ABCG8, and MDR3 messenger RNA (mRNA) were strongly correlated. After transplantation, the biliary secretion rate of cholesterol continuously increased, coinciding with gradual increases in bile salt and phospholipid secretion. In contrast, hepatic levels of ABCG5 and ABCG8 mRNA remained unchanged. Surprisingly, no correlation was found between the hepatic expression of ABCG5 and ABCG8 and rates of biliary cholesterol secretion, normalized for biliary phospholipid secretion. As expected, the concentration of biliary phospholipids correlated well with MDR3 expression. In conclusion, the strong relationship between ABCG5 and ABCG8 gene expression is consistent with the coordinate regulation of both genes, and in line with heterodimerization of both proteins into a functional transporter. Hepatic ABCG5/ABCG8 expression, at least during the early phase after transplantation, is not directly related to biliary cholesterol secretion in humans. This finding suggests the existence of alternative pathways for the hepatobiliary transport of cholesterol that are not controlled by ABCG5/ABCG8. PMID- 16250036 TI - In type 1 autoimmune hepatitis, is cirrhosis at presentation or follow-up associated with a poorer outcome? PMID- 16250037 TI - Lamivudine plus interleukin-12 combination therapy in chronic hepatitis B: antiviral and immunological activity. AB - Interleukin-12 (IL-12) is an immunomodulatory cytokine that promotes cellular immunity. Pre-clinical data suggest that IL-12 inhibits hepatitis B virus (HBV) replication by stimulating interferon-gamma (IFN-gamma) production. We investigated whether a combination treatment with lamivudine plus recombinant human interleukin-12 (rhIL-12) will result in a greater and prolonged suppression of HBV replication in comparison with lamivudine monotherapy. Fifteen patients with HBeAg-positive chronic hepatitis B were randomized to receive either lamivudine alone for 24 weeks (group 1); combination of lamivudine for 16 weeks and rhIL-12 (200 ng/kg twice weekly), starting 4 weeks after initiation of lamivudine, for 20 weeks (group 2), or the same schedule as for group 2, with lamivudine and a higher dose of rhIL-12 (500 ng/kg, group 3). Serum HBV DNA levels, T-cell proliferation, frequency of virus-specific T-cells, and IFN-gamma production were evaluated serially during and 24 weeks posttreatment. Lamivudine plus rhIL-12/500 showed greater antiviral activity than lamivudine monotherapy. However, after stopping lamivudine in groups 2 and 3, serum HBV DNA increased significantly despite continuing rhIL-12 administration. Lamivudine plus rhIL-12 treatment was associated with a greater increase in virus-specific T-cell reactivity, IFN-gamma production, and an inverse correlation between the frequency of IFN-gamma-producing CD4+ T-cells and viremia. The T-cell proliferative response to HBcAg did not differ between the three groups. In conclusion, the addition of IL-12 to lamivudine enhances T-cell reactivity to HBV and IFN-gamma production. However, IL-12 does not abolish HBV replication in HBeAg-positive patients and does not maintain inhibition of HBV replication after lamivudine withdrawal. PMID- 16250039 TI - Methotrexate (MTX) plus ursodeoxycholic acid (UDCA) in the treatment of primary biliary cirrhosis. AB - This placebo-controlled, randomized, multicenter trial compared the effects of MTX plus UDCA to UDCA alone on the course of primary biliary cirrhosis (PBC). Two hundred and sixty five AMA positive patients without ascites, variceal bleeding, or encephalopathy; a serum bilirubin less than 3 mg/dL; serum albumin 3 g/dL or greater, who had taken UDCA 15 mg/kg daily for at least 6 months, were stratified by Ludwig's histological staging and then randomized to MTX 15 mg/m2 body surface area (maximum dose 20 mg) once a week while continuing on UDCA. The median time from randomization to closure of the study was 7.6 years (range: 4.6-8.8 years). Treatment failure was defined as death without liver transplantation; transplantation; variceal bleeding; development of ascites, encephalopathy, or varices; a doubling of serum bilirubin to 2.5 mg/dL or greater; a fall in serum albumin to 2.5 g/dL or less; histological progression by at least two stages or to cirrhosis. Patients were continued on treatment despite failure of treatment, unless transplantation ensued, drug toxicity necessitated withdrawal, or the patient developed a cancer. There were no significant differences in these parameters nor to the time of development of treatment failures observed for patients taking UDCA plus MTX, or UDCA plus placebo. The trial was conducted with a stopping rule, and was stopped early by the National Institutes of Health at the advice of our Data Safety Monitoring Board for reasons of futility. In conclusion, methotrexate when added to UDCA for a median period of 7.6 years had no effect on the course of PBC treated with UDCA alone. PMID- 16250041 TI - What happens to cirrhotic cardiomyopathy after liver transplantation? PMID- 16250040 TI - Risk factors and comorbidities in primary biliary cirrhosis: a controlled interview-based study of 1032 patients. AB - Primary biliary cirrhosis (PBC) is an autoimmune disease of unknown etiology, often associated with other autoimmune conditions. Controlled studies have so far provided conflicting data on risk factors and comorbidity rates in PBC. We enrolled patients with PBC (n = 1032) from 23 tertiary referral centers for liver diseases in the United States and random-digit-dialed controls (n = 1041) matched for sex, age, race, and geographical location. Patients and controls were administered a modified version of the US National Health and Nutrition Examination Study (NHANES III) questionnaire by trained personnel to evaluate associations between PBC and social, demographic, personal and family medical histories, lifestyle, and reproductive factors and the rates of comorbidity in affected individuals. Data indicate that having a first-degree relative with PBC (adjusted odds ratio [AOR] 10.736; 95% confidence interval 4.227-27.268), history of urinary tract infections (AOR 1.511, 95% CI 1.192-1.915), past smoking (AOR 1.569, 95% CI 1.292-1.905), or use of hormone replacement therapies (AOR 1.548, 95% CI 1.273-1.882) were significantly associated with increased risk of PBC. The frequent use of nail polish slightly increased the risk of having PBC. Other autoimmune diseases were found in 32% of cases and 13% of controls (P<0.0001). In conclusion, environmental factors, possibly including infectious agents through urinary tract infections or chemicals contained in cigarette smoke, may induce PBC in genetically susceptible individuals. Exogenous estrogens may also contribute to explain the female predominance of the disease. PMID- 16250042 TI - Placebo controlled treatment trials in hepatic encephalopathy are feasible and ethical. PMID- 16250043 TI - Insulin resistance: a metabolic pathway to chronic liver disease. AB - Insulin resistance (IR) is the pathophysiological hallmark of nonalcoholic fatty liver disease (NAFLD), one of the most common causes of chronic liver disease in Western countries. We review the definition of IR, the methods for the quantitative assessment of insulin action, the pathophysiology of IR, and the role of IR in the pathogenesis of chronic liver disease. Increased free fatty acid flux from adipose tissue to nonadipose organs, a result of abnormal fat metabolism, leads to hepatic triglyceride accumulation and contributes to impaired glucose metabolism and insulin sensitivity in muscle and in the liver. Several factors secreted or expressed in the adipocyte contribute to the onset of a proinflammatory state, which may be limited to the liver or more extensively expressed throughout the body. IR is the common characteristic of the metabolic syndrome and its related features. It is a systemic disease affecting the nervous system, muscles, pancreas, kidney, heart, and immune system, in addition to the liver. A complex interaction between genes and the environment favors or enhances IR and the phenotypic expression of NAFLD in individual patients. Advanced fibrotic liver disease is associated with multiple features of the metabolic syndrome, and the risk of progressive liver disease should not be underestimated in individuals with metabolic disorders. Finally, the ability of insulin sensitizing, pharmacological agents to treat NAFLD by reducing IR in the liver (metformin) and in the periphery (thiazolidinediones) are discussed. PMID- 16250044 TI - Trimming the fat from liver regeneration. PMID- 16250045 TI - Infection of human hepatocyte chimeric mouse with genetically engineered hepatitis B virus. AB - Studies of hepatitis B virus (HBV) mutants have been hampered by the lack of a small animal model with long-term infection of cloned HBV. Using a mouse model in which liver cells were highly replaced with human hepatocytes that survived over a long time with mature human hepatocyte function, we performed transmission experiments of HBV. Human serum containing HBV and the virus produced in HepG2 cell lines that transiently or stably transfected with 1.4 genome length HBV DNA were inoculated. Genetically modified e-antigen-negative mutant strain also was produced and inoculated into the mouse model. A high-level (approximately 10(10) copies/mL) viremia was observed in mice inoculated with HBV-positive human serum samples. The level of viremia tended to be high in mice with a continuously high human hepatocyte replacement index. High levels and long-lasting viremia also were observed in mice injected with the in vitro generated HBV. The viremia continued up to 22 weeks until death or killing. Passage experiments showed that the serum of these mice contained infectious HBV. Genetically engineered hepatitis B e antigen-negative mutant clone also was shown to be infectious. Lamivudine effectively reduced the level of viremia in these infected mice. In conclusion, this mouse model of HBV infection is a useful tool for the study of HBV virology and evaluation of anti-HBV drugs. Our results indicate that HBeAg is dispensable for active viral production and transmission. PMID- 16250046 TI - Lack of gp130 expression results in more bacterial infection and higher mortality during chronic cholestasis in mice. AB - Chronic cholestasis is associated with increased bacterial infections and sepsis resulting in higher mortality in humans. In the current study, we investigated the relevance of gp130-dependent pathways after bile duct ligation (BDL). BDL was performed in conditional gp130 knockout (loxP/Cre system) mice and respective controls. Liver injury, regulation of the acute phase response, and the impact on survival and bacterial infections were determined. Acute BDL resulted in increased IL-6 levels, Stat3 activation, and an increase in acute-phase proteins (serum-amyloid-A [SAA]), which was blocked in gp130-deleted animals. In addition, the antimicrobial gene hepcidin was regulated in a gp130-dependent manner after BDL. During chronic cholestasis Stat3 activation was dramatically reduced, while high SAA levels were maintained via gp130-dependent signaling. Inhibition of gp130-dependent pathways resulted in higher mortality and liver damage, which was associated with higher infiltration of immune-activated cells and increased germ number in the liver. In conclusion, during acute and chronic cholestasis, the gp130 system is essential for controlling the acute-phase response. Lack of gp130 expression results in pronounced bacterial growth in bile and liver after BDL, which is associated with higher mortality. Activation of gp130-dependent pathways after BDL is essential and appears to be a therapeutic target during cholestasis. PMID- 16250047 TI - Reflections on therapeutic trials in primary biliary cirrhosis. PMID- 16250048 TI - Human monoclonal antibodies that react with the E2 glycoprotein of hepatitis C virus and possess neutralizing activity. AB - Active and/or passive immunoprophylaxis against hepatitis C virus (HCV) remain unachieved goals. Monoclonal antibodies might provide one approach to protection. We derived human monoclonal antibodies from the bone marrow of a patient with a well-controlled HCV infection of 22 years duration. Five distinct antibodies reactive with the E2 glycoprotein of the homologous 1a strain of HCV were recovered as antigen-binding fragments (FAbs). They demonstrated affinity constants as high as 2 nanomolar. "Neutralization of binding" titers paralleled the affinity constants. All five FAbs reacted with soluble E2 protein only in nonreducing gels, indicating that the relevant epitopes were conformational. The FAbs could be divided into two groups, based on competition analysis. Three of the FAbs neutralized the infectivity of pseudotyped virus particles (pp) bearing the envelope glycoproteins of the homologous HCV strain (genotype 1a). The three FAbs also neutralized genotype 1b pp and one also neutralized genotype 2a pp. In conclusion, one or more of these monoclonal antibodies may be useful in preventing infections by HCV belonging to genotype 1 or 2, the most medically important genotypes worldwide. PMID- 16250049 TI - Early intrahepatic antigen-specific retention of naive CD8+ T cells is predominantly ICAM-1/LFA-1 dependent in mice. AB - We have previously shown that naive CD8+ T cells recognizing their cognate antigen within the liver are retained and undergo activation in situ, independent from lymphoid tissues. Intrahepatic primary T cell activation results in apoptosis and may play a crucial role in the ability of the liver to induce tolerance. Although adhesion molecules required for intrahepatic retention of T cells that have undergone previous extra-hepatic activation have been characterized, adhesive interactions involved in selective antigen-dependent intrahepatic retention of naive CD8+ T cells have not been investigated. By adoptively transferring radiolabeled T cell receptor (TCR)-transgenic CD8+ T cells into recipient animals ubiquitously expressing the relevant antigen, we show that 40% to 60 % of donor antigen-specific naive CD8+ T cells were retained in the liver within 1 hour after transfer, despite ubiquitous expression of the antigen. Intravital microscopy showed that most donor naive T cells slowed down and were irreversibly retained intrahepatically within the first few minutes after adoptive transfer, strongly suggesting that they were directly activated by liver cells in situ. This process was largely dependent on LFA-1 and ICAM-1, but was independent of blocking with antibodies against VCAM-1, alpha4 integrin, P selectin, VAP-1, and beta1 integrin. ICAM-2 seemed to play only a minor role in this process. Interestingly, LFA-1 expressed by both donor T cells and liver cells was involved in retention of the antigen-reactive T cells. In conclusion, LFA-1-dependent intrahepatic T cell retention and activation are linked events that may play a crucial role in the establishment of liver-induced antigen specific tolerance. PMID- 16250050 TI - Altered disposition of acetaminophen in mice with a disruption of the Mrp3 gene. AB - MRP3 is an ABC transporter localized in the basolateral membrane of epithelial cells such as hepatocytes and enterocytes. In this study, the role of Mrp3 in drug disposition was investigated. Because Mrp3 preferentially transports glucuronide conjugates, we investigated the in vivo disposition of acetaminophen (APAP) and its metabolites. Mrp3+/+ and Mrp3-/- knockout mice received APAP (150 mg/kg), and bile was collected. Basolateral and canalicular excretion of APAP was also assessed in the isolated perfused liver. In separate studies, mice received 400 mg APAP/kg for assessment of hepatotoxicity. No differences were found in the biliary excretion of APAP, APAP-sulfate, and APAP-glutathione between Mrp3+/+ and Mrp3-/- mice. However, 20-fold higher accumulation of APAP-glucuronide (APAP GLUC) was found in the liver of Mrp3-/- mice. Concomitantly, plasma APAP-GLUC content in Mrp3-/- mice was less than 10% of that in Mrp3+/+ mice. In addition, APAP-GLUC excretion in bile of Mrp3-/- mice was tenfold higher than in Mrp3+/+ mice. In the isolated perfused liver, we also found a strong decrease of APAP GLUC secretion into the perfusate of Mrp3-/- livers. Plasma alanine aminotransferase (ALT), aspartate aminotransferase (AST), and histopathology showed that Mrp3-/- mice are more resistant to APAP hepatotoxicity than Mrp3+/+ mice, which is most likely a result of the faster repletion of hepatic GSH. In conclusion, basolateral excretion of APAP-GLUC in mice is nearly completely dependent on the function of Mrp3. In its absence, sufficient hepatic accumulation occurs to redirect some of the APAP-GLUC to bile. This altered disposition in Mrp3-/- mice is associated with reduced hepatotoxicity. PMID- 16250051 TI - Management of hepatocellular carcinoma. PMID- 16250053 TI - Ethanol metabolism alters interferon gamma signaling in recombinant HepG2 cells. AB - We previously showed that IFNgamma signal transduction was suppressed by ethanol in recombinant HepG2 cells (VL-17A cells), which express alcohol dehydrogenase (ADH) and CYP2E1. We examined the mechanisms by which STAT1 phosphorylation is blocked by ethanol treatment in VL-17A cells. Cells were exposed to 0 or 100 mmol/L ethanol for 72 hours. STAT1 phosphorylation was determined by Western blot after 1 hour IFNgamma exposure. Reduction of STAT1 phosphorylation by ethanol was prevented in the presence of 4MP, DAS, or uric acid, indicating that the oxidative products from ethanol metabolism were partly responsible for suppression of STAT1 phosphorylation. Ethanol exposure decreased STAT1 tyrosine phosphorylation, whereas serine phosphorylation on the protein was unchanged. These effects of ethanol were mimicked by the peroxynitrite (PN) donor, SIN-1, which also blocked tyrosine, but not serine phosphorylation, on STAT1. When cells expressing either ADH (VA-13 cells) or CYP2E1 (E-47 cells) were exposed to ethanol, both ADH- and CYP2E1-generated products reduced STAT1 phosphorylation. In addition, SOCS1, a negative regulator of IFNgamma signaling and which is degraded by the proteasome, was stabilized by ethanol treatment, presumably because of inhibited proteasome activity. Furthermore, SIN-1 treatment elevated SOCS1 levels in VL-17A cells, indicating that PN has a role in SOCS1 elevation. In conclusion, under conditions of ethanol-elicited oxidative stress, PN prevents STAT1 phosphorylation by stabilization of SOCS1, and possibly by nitration of tyrosine residues in STAT1 protein. PMID- 16250054 TI - Transcriptional profiling after bile duct ligation identifies PAI-1 as a contributor to cholestatic injury in mice. AB - Extrahepatic cholestasis leads to complex injury and repair processes that result in bile infarct formation, neutrophil infiltration, cholangiocyte and hepatocyte proliferation, extracellular matrix remodeling, and fibrosis. To identify early molecular mechanisms of injury and repair after bile duct obstruction, microarray analysis was performed on liver tissue 24 hours after bile duct ligation (BDL) or sham surgery. The most upregulated gene identified encodes plasminogen activator inhibitor 1 (PAI-1, Serpine 1), a protease inhibitor that blocks urokinase plasminogen activator (uPA) and tissue-type plasminogen activator (tPA) activity. Because PAI-1, uPA, and tPA influence growth factor and cytokine processing as well as extracellular matrix remodeling, we evaluated the role of PAI-1 in cholestatic liver injury by comparing the injury and repair processes in wild type (WT) and PAI-1-deficient (PAI-1-/-) mice after BDL. PAI-1-/- mice had fewer and smaller bile infarcts, less neutrophil infiltration, and higher levels of cholangiocyte and hepatocyte proliferation than WT animals after BDL. Furthermore, PAI-1-/- mice had higher levels of tPA activation and mature hepatocyte growth factor (HGF) after BDL than WT mice, suggesting that PAI-1 effects on HGF activation critically influence cholestatic liver injury. This was further supported by elevated levels of c-Met and Akt phosphorylation in PAI-1-/- mice after BDL. In conclusion, PAI-1 deficiency reduces liver injury after BDL in mice. These data suggest that inhibiting PAI-1 might attenuate liver injury in cholestatic liver diseases. PMID- 16250055 TI - Temporal correlation of pathology and DNA damage with gene expression in a choline-deficient model of rat liver injury. AB - Hepatocellular carcinoma (HCC) is the terminal event in chronic liver diseases with repeated cycles of cellular injury and regeneration. Although much is known about the cellular pathogenesis and etiological agents leading to HCC, the molecular events are not well understood. The choline-deficient (CD) model of rodent HCC involves the consecutive emergence of a fatty liver, apoptosis, compensatory proliferation, fibrosis, and cirrhosis that is markedly similar to the sequence of events typified by human HCC. Moreover, oxidative stress is thought to play a pivotal role in the progression of the disease. Here, we hypothesize that gene expression profiling can temporally mirror the histopathology and oxidative DNA damage observed with this model. We show that clusters of highly co-regulated genes representing distinct cellular pathways for lipid biosynthesis and metabolism, apoptosis, cell proliferation, and tissue remodeling temporally correlate with the well-defined sequential emergence of pathological alterations in the progression of liver disease. Additionally, an oxidative stress signature was observed that was corroborated in a time-dependent manner with increases in oxidized purines and abasic sites in DNA. Collectively, expression patterns were strongly driven by pathology, demonstrating that patterns of gene expression in advanced stages of liver disease are primarily driven by histopathological changes and to a much lesser degree by the original etiological agent. In conclusion, gene expression profiling coupled with the CD model of HCC provides a unique opportunity to unveil the molecular events associated with various stages of liver injury and carcinogenesis and to distinguish between causal and consecutive changes. PMID- 16250056 TI - Spectroscopic Studies of water-soluble porphyrins with protein encapsulated in bis(2-ethylhexyl)sulfosuccinate (AOT) reverse micelles: aggregation versus complexation. AB - We have investigated the interaction of two water-soluble free-base porphyrins (negatively charged meso-tetrakis(p-sulfonatophenyl)porphyrin sodium salt (TSPP) and positively charged meso-tetrakis(N-methylpyridinium-4-yl)porphyrin (TMpyP)) with two drug-carrier proteins (human serum albumin (HSA) and beta-lactoglobulin (betaLG)) in bis(2-ethylhexyl)sulfosuccinate (AOT)/isooctane/water reverse micelles (RM) by using steady-state and transient-state fluorescence spectroscopy. TSPP exhibited a complex pattern of aggregation on varying the RM size and pH in the absence of the protein: at low omega0 (the ratio of water concentration to AOT concentration, the emission of H-aggregates prevails under acidic or neutral "pH(ext)" conditions. Upon formation of the water-pool, J aggregates and monomeric diacid species dominate at low "pH(ext)" but only monomer is detected at neutral "pH(ext)". The aggregation number increases with omega0 and the presence of the protein does not seem to contribute to further growth of the aggregate. The presence of protein leads to H-deaggregation but promotes J-aggregation up to a certain protein/porphyrin ratio above which, complexation with the monomer bound to a hydrophobic site of the protein prevails. The effective complex binding constants are smaller than in free aqueous solution; this indicates a weaker binding in these RM probably due to some conformational changes imposed by encapsulation. Only a weak quenching of TMpyP fluorescence is detected due to the presence of protein in contrast to the negative porphyrin. PMID- 16250057 TI - Electro- and magnetocommunication in [5,5]ditrovacenyls, [(eta7-C7H7)V(eta5-C5H4 X-eta5-C5H4)V(eta7-C7H7)], mediated by the spacers X = (Z)-CH=CH-, (E)-CH=CH-, > C=CH2, -CH2CH2-, and -CH2-. AB - Five new paramagnetic dinuclear complexes containing [5]trovacenyl groups, (eta7 C7H7)V(eta5-C5H4-), have been prepared and characterized, including by single crystal X-ray diffraction. As intervening spacers, ethenediyl units in the geminal and vicinal (Z)- and (E)-bridging modes as well as methanediyl and ethanediyl units have been included with the aim of studying their propensity to transmit electric and magnetic information. It is found that redox splitting of consecutive electron-transfer steps is resolved for reduction (0-->1- -->2-) only, unsaturation of the C2 bridge not being requisite, since the -CH2CH2- spacer also gives rise to a small redox splitting. Magnetic communication is quantified in terms of the exchange coupling constant J, accessible from the EPR hyperfine pattern in solution and from magnetic susceptometry in the solid state. The results obtained from these methods generally differ; this fact is not surprising in view of conformational differences in the respective states of aggregation. It is concluded that orientation-dependent mechanisms of spin-spin interactions (pi-orbital overlap, hyperconjugation) contribute extensively although, as implied by sizeable J values for -CH2- and -C2H4- linked di[5]trovacenyl groups, coupling mediated by the sigma-orbital chain must also be considered. PMID- 16250058 TI - Controlled nanozeolite-assembled electrode: remarkable enzyme-immobilization ability and high sensitivity as biosensor. AB - An enzyme-immobilized nanozeolite-assembled electrode was prepared by controlled assembly of nanometer-sized Linder type-L zeolite (nano-LTL-zeolite) on an indium tin oxide (ITO) glass electrode surface, and subsequent immobilization of cytochrome c. Cyclic voltammetric (CV) and amperometric experiments showed that, relative to other reported electrodes, the enzyme-immobilized electrodes possess fast electron-transfer rates (2.2 s(-1)), a broad linear range (15-540 micromol L(-1)), a low detection limit (3.2 nmol L(-1)), a remarkably long lifetime (5 months), and high stability in the pH range 5-10. These characteristics could be due to the fact that nanozeolites assembled on ITO have high immobilization ability and facilitate interaction with enzymes. The function controllability of these enzyme electrodes, resulting from the facile manipulability of nanozeolite assembled layers, may provide a possibility to rationally design biosensors. PMID- 16250059 TI - A competitive molecular recognition study: syntheses and analysis of supramolecular assemblies of 3,5-dihydroxybenzoic acid and its bromo derivative with some N-donor compounds. AB - A molecular recognition study of 3,5-dihydroxybenzoic acid (1) and its bromo derivative 4-bromo-3,5-dihydroxybenzoic acid (2) with the N-donor compounds 1,2 bis(4-pyridyl)ethene (bpyee), 1,2-bis(4-pyridyl)ethane (bpyea), and 4,4' bipyridine (bpy) is reported. Thus, the syntheses and structural analysis of molecular adducts 1 a-1 c (1 with bpyee, bpyea, and bpy, respectively) and 2 a-2 c (2 with bpyee, bpyea, and bpy, respectively) are discussed. In all these adducts, recognition between the constituents is established through either O- H...N and/or O--H...N/C--H...O pairwise hydrogen bonds. In all the adducts both OH and COOH functional groups available on 1 and 2 interact with the N-donor compounds, except in 2 a, in which only COOH (COO-) is involved in the recognition process. The COOH moieties in 1 a, 1 b, and 2 b form only single O- H...N hydrogen bonds, whereas in 1 c and 2 c, they form pairwise O--H...N/C- H...O hydrogen bonds. In addition, subtle differences in the recognition patterns resulted in the formation of cyclic networks of different dimensions. In fact, only 1 c forms a four-molecule cyclic moiety, as was already documented in the literature for this kind of assemblies. All complexes have been characterized by single-crystal X-ray diffraction. The supramolecular architectures are quite elegant and simple, with stacking of sheets in all adducts, but a rather complex network with a threefold interpenetration pattern was found in 2 c. PMID- 16250060 TI - The transfer of tin and germanium atoms from N-heterocyclic stannylenes and germylenes to diazadienes. AB - New N-heterocyclic stannylenes and germylenes were synthesized by transamination of E[N(SiMe3)2] (E = Ge, Sn) with alpha-amino-aldimines or ethylidene-1,2 diamines and were characterized by spectroscopic methods and in the case of the germylene 10 g by X-ray diffraction. The reactions of several germylenes and stannylenes with diazadienes were studied by using dynamic NMR and computational methods. Experimental and theoretical studies confirmed that metathesis with exchange of the Group 14 atom is feasible for both stannylenes and germylenes, with exchange rates being generally higher for stannylenes. The metathesis of the diazadiene 3 b and the stannylene 1 b follows second-order kinetics and exhibits a sizeable negative entropy of activation. The transfer reaction is inhibited by bulky substituents in both reactants and surprisingly coincides with a suppression of the fragmentation of the stannylene into tin and diazadiene. A connection between oxidative addition and ring fragmentation was also observed in the reaction of 1 f with sulfur. Density functional theory (DFT) calculations suggest that all metathesis reactions proceed via transient spirocyclic [1+4] cycloaddition products, the formation of which is generally endothermic and endergonic. The spirostannanes display a distorted Psi-tbp geometry at the tin atom and their cycloreversion requires low or nearly negligible activation energies; spirogermanes exhibit distorted tetrahedral central atoms and sizeable energy barriers with respect to the same reaction. Complementary studies of cycloadditions of diazadienes to triplet germylenes or stannylenes indicate that these reactions are exothermic. The lowest triplet state in the carbene homologues results from promotion of an electron from an n(N) orbital with pi character rather than the n(C)-sigma orbital as in carbenes, and singlet-triplet excitation energies decrease from carbon to tin. Spirostannanes exhibit a triplet ground-state multiplicity that implies that the energy hypersurfaces for the reactions of singlet and triplet stannylenes with diazadienes intersect; for germylenes, the singlet hypersurface is always lower in energy. A reaction mechanism explaining the different thermal stabilities of N-heterocyclic germylenes and stannylenes, and the coincidence between ring metathesis and thermal decomposition of the latter, is proposed based on the different separation of the singlet and triplet energy hypersurfaces. PMID- 16250061 TI - Synthesis of functionalized vinylgermanes through a new ruthenium-catalyzed coupling reaction. AB - Vinyl-substituted germanes react stereo- and regioselectively with olefins in the presence of complexes containing Ru-H and Ru-Ge bonds with the formation of functionalized vinylgermanes that cannot be synthesized by olefin cross metathesis procedures. The reaction opens a new catalytic route for preparation of a class of organogermanes that are potent organometallic reagents for organic synthesis because they show very low toxicity and could replace organotin compounds. The mechanism of this new catalytic route was proven to involve an interesting insertion of the vinylgermane into the Ru-H bond and beta-Ge transfer to the metal with elimination of ethylene and generation of an Ru-Ge bond, followed by insertion of the alkene into the Ru-Ge bond and beta-H transfer to the metal to eliminate the substituted vinylgermane. PMID- 16250062 TI - Structural preferences of single-walled silica nanostructures: nanospheres and chemically stable nanotubes. AB - Structural preferences of single-walled and coordinatively saturated spherical and tubular nanostructures of silica have been determined by ab initio calculations. Two families of spherical (SiO2)n clusters derived from Platonic solids and Archimedean polyhedra are depicted, with n ranging from 4-120. The analogue of a truncated icosidodecahedron, Ih-symmetric Si120O240, is favored in energy, closely followed by the Ih-symmetric Si60O120-truncated icosahedron. The silica nanotubes derived from spherical clusters are capped by Si2O2 rings, whereas the tubular section consists of single oxygen bridges. Periodic studies performed with open-ended silica nanotubes and the alpha-quartz polymorph of silica, along with a comparisons to fullerenes and carbon nanotubes, suggest that tubes with diameters of approximately 1 nm should be chemically stable. PMID- 16250063 TI - Ruthenium complexes with the stanna-closo-dodecaborate ligand: coexistence of eta1(Sn) and eta3(B-H) coordination. AB - Four stanna-closo-dodecaborate complexes of ruthenium have been prepared and characterized by multinuclear NMR studies in solution and in the solid state. The solid-state structures of the dimeric zwitterions [[Ru(dppb)(SnB11H11)]2] (2) (dppb = bis(diphenylphosphino)butane), [[Ru(PPh3)2(SnB11H11)]2] (3), and the dianionic ruthenium complex [Bu3MeN]2[Ru(dppb)[2,7,8-(mu-H)3-exo SnB11H11](SnB11H11)] (4) were determined by X-ray crystal structure analysis; they establish an unprecedented structural motif in the chemistry of heteroboranes and transition-metal fragments with the stanna-closo-dodecaborate moiety as a two-faced ligand that exhibits eta1(Sn) as well as eta3(B-H) coordination. The eta3-coordinated stannaborate in 4 and in the isostructural compound [Bu3MeN]2[Ru(PPh3)2[2,7,8-(mu-H)3-exo-SnB11H11](SnB11H11)] (5) shows fluxional behavior, which was studied in detail by using 31P[1H] EXSY and DNMR experiments. The activation parameters for the dynamic process of 5 are given. PMID- 16250064 TI - Neuropsychiatric symptoms of dementia: cross-sectional analysis from a prospective, longitudinal Belgian study. AB - OBJECTIVE: Given the rather limited knowledge on profiles of neuropsychiatric symptoms (behavioural and psychological signs and symptoms of dementia, BPSD) in several degenerative dementias, we designed a prospective study of which we here present the baseline data. METHODS: Diagnosed according to strictly applied clinical diagnostic criteria, patients with probable Alzheimer's disease (AD) (n = 205), frontotemporal dementia (FTD) (n = 29), mixed dementia (MXD) (n = 39) and dementia with Lewy bodies (DLB) (n = 23) were included. All patients underwent a neuropsychological examination and behavioural assessment by means of a battery of scales (Middelheim Frontality Score (MFS), Behave-AD, Cohen-Mansfield Agitation Inventory, Cornell Scale for Depression in Dementia). RESULTS: In AD and MXD, activity disturbances and aggressiveness occurred in more than 80% of the patients. With a prevalence of 70%, apathy was very common whereas delusions and hallucinations were rare in FTD patients. Frequently used behavioural assessment scales like the Behave-AD systematically underestimated BPSD in FTD whereas the MFS displayed high sensitivity for frontal lobe symptoms. Hallucinations discriminated DLB patients from other dementias. A high prevalence of disinhibition (65%) in DLB pointed to frontal lobe involvement. CONCLUSIONS: Behavioural assessment may help differentiating between different forms of dementia, further stressing the need for the development of new and more sensitive behavioural assessment scales. By means of the MFS, frontal lobe involvement was frequently observed in DLB. As 70% of FTD patients displayed apathy, prevalence was about two times higher compared to the other disease groups, meanwhile indicating that apathy is frequently observed in dementia, irrespective of its etiology. PMID- 16250067 TI - Investigation of the mechanism of resistance to third-generation cephalosporins by class C beta-lactamases by using chemical complementation. AB - The widespread use of antibiotics to treat bacterial infections has led to the continuing challenge of antibiotic resistance. For beta-lactam antibiotics, the most common form of resistance is the expression of beta-lactamase enzymes, which inactivate the antibiotics by cleavage of the beta-lactam core. In this study, chemical complementation, which is a general method to link the formation or cleavage of a chemical bond to the transcription of a reporter gene in vivo, was employed in combination with combinatorial mutagenesis to study the mechanism by which the class C beta-lactamase P99 might evolve resistance to the commonly administered third-generation cephalosporin cefotaxime. The chemical complementation system was first shown to be able to distinguish between the wild type (wt) class C beta-lactamase P99 and the clinically isolated extended spectrum class C beta-lactamase GC1 in the presence of cefotaxime. The system was then employed to evaluate the activity of mutants of wt P99 towards cefotaxime. A number of single-point mutations at position 221 (Tyr in wt P99) were identified that conferred resistance towards inhibition by cefotaxime, with as much as a 2000-fold increase in k(cat) and a 100-fold increase in k(cat)/K(M) (k(cat)=the rate of catalysis; K(M)=the Michaelis constant), as compared to those of the wt enzyme. Finally, the chemical complementation system was employed in a high throughput screen to identify a number of mutants of P99 that have multiple mutations around the substrate-binding pocket that increase resistance towards cefotaxime inhibition. The catalytic turnover of cefotaxime by the most active mutant identified was 5500 times higher than that of the wt P99. The resistant mutants suggest a mechanism by which a number of mutations can confer resistance by increasing the flexibility of the Omega loop and altering the positioning of residue 221. Thus, as illustrated in this study, chemical complementation has the potential to be used as a high-throughput screen to study a wide range of enzyme drug interactions. PMID- 16250070 TI - The utility of EEG in dementia: a clinical perspective. AB - BACKGROUND: Despite being simple and cheap, the EEG is not often used in clinical practice. METHODOLOGY: Literature search using PUBMED and Medline. RESULTS: Quantitative EEG can help to identify mild dementia and mild cognitive impairment and can increase diagnostic accuracy when used with other imaging techniques. EEG helps differentiate organic from functional brain disease and predict response to cholinesterase inhibitors and is central in the diagnosis of Creutzfeldt Jacob disease. The accuracy of EEG may be greater than that of CT or MRI scans alone. DISCUSSION: Quantitative EEG may save on specialist interpretation time and enable more routine use of EEG in diagnosis and care. More widespread use of EEG's is indicated. Agreement on the parameters that are best measured on qEEG is still awaited. PMID- 16250069 TI - Olanzapine does not enhance cognition in non-agitated and non-psychotic patients with mild to moderate Alzheimer's dementia. AB - OBJECTIVE: This was an exploratory study of olanzapine as potential treatment for improvement in cognition in patients with Alzheimer's disease without prominent psychobehavioral symptoms. METHODS: Non-psychotic/non-agitated patients (n = 268) with Alzheimer's disease, who had baseline Mini-Mental State Examination (MMSE) scores of 14-26 were randomized to treatment with olanzapine (2.5 to 7.5 mg/d) or placebo for 26 weeks. The primary objectives were to determine if treatment with olanzapine improved cognition as indexed by the Alzheimer's disease Assessment Scale for Cognition (ADAS-Cog) and the Clinician's Interview-Based Impression of Change (CIBIC) after 26 weeks of therapy. RESULTS: Patients treated with olanzapine vs placebo experienced significant worsening ADAS-Cog scores at weeks 12 (p = 0.03) and 26 (p = 0.004). Changes in CIBIC scores were not significantly different between treatment groups at either assessment. A post hoc analysis revealed that olanzapine-treated patients with more cognitive impairment at baseline (MMSE scores of 14-18) (n = 35) experienced significantly greater deterioration in ADAS-Cog performance than patients in the placebo group (n = 24; p < 0.001); whereas in patients with less cognitive impairment (n = 78, baseline MMSE scores of 23-26) between-group ADAS-Cog changes were not significant. CONCLUSIONS: In this 26-week study non-psychotic/non-agitated patients with Alzheimer's disease treated with olanzapine experienced significant worsening of cognition as compared to placebo. PMID- 16250071 TI - Mitochondrial aldehyde dehydrogenase polymorphism is not associated with incidence of Alzheimer's disease. AB - BACKGROUND: The association between the mutant allele of mitochondrial aldehyde dehydrogenase (ALDH2*2) and Alzheimer's disease (AD) has been controversial and only so far investigated in cross-sectional studies. This study aimed to investigate the longitudinal association between ALDH2*2 and incidence of AD. METHODS: Of 592 participants aged 65 or over without dementia at baseline, 510 (86%) were re-evaluated after 2.4 years and comprised the study sample. Baseline measures included: demographic characteristics, drinking behavior, cognitive function (MMSE), clinical diagnoses of dementia and AD, and genotype (ALDH2 and apolipoprotein E). At the follow up examination, alcohol related characteristics, MMSE and clinical diagnoses of dementia and AD were reassessed. RESULTS: There were no significant associations between the ALDH2*2 and any cognitive outcomes (incidence of dementia or AD, or cognitive decline). These findings were not changed after adjustment for alcohol consumption. No interaction was found between ALDH2 and apolipoprotein E. CONCLUSIONS: ALDH2*2 does not seem to be important in the etiology of dementia. PMID- 16250074 TI - Sensitivity of somatic symptoms in post-stroke depression (PSD). PMID- 16250072 TI - Provision of care for older people with co-morbid mental illness in general hospitals: general nurses' perceptions of their training needs. AB - INTRODUCTION: There are high levels of co-morbid mental illness amongst older people in general hospitals; this study explored the training needs of general nurses to care for this group. METHOD: Focus groups with general nurses were analysed using framework analysis. FINDINGS AND CONCLUSION: Nurses wanted training, but did not believe that training alone was sufficient to improve care, expressing that more integrated working between acute and mental health services was also needed. Liaison mental health services provide a way to deliver both training and a more integrated service. PMID- 16250075 TI - Gender differences in the contributions of risk factors to depressive symptoms among the elderly persons dwelling in a community, Japan. AB - OBJECTIVE: To examine the relative importance of risk factors associated with depressive symptoms and gender differences in exposure to the risk factors among the elderly persons living in the community. METHODS: The data came from the Minamifurano-town Aging Study, a community-based sample of non-institutionalized elderly persons aged 65 years or older. Of the 731 eligible subjects, 665 were assessed for four domains of the potential risk factors (demographic characteristics, health and disability, stress, and social networks) and depressive symptoms according to the 30-item Geriatric Depression Scale (GDS). RESULTS: The mean overall GDS-score was 10.9 (SD 6.2), 10.2 (SD 6.0) in men and 11.6 (SD 6.4) in women. The stress domain in men and the health and disability domain in women contributed most to the explanation of the variation in the GDS score. CONCLUSION: 'Stress' for men and 'health and disability status' for women were important factors associated with depressive symptoms. Future studies should determine whether modification of these factors may prevent depression among the elderly persons living in the community. PMID- 16250076 TI - Migrant status, age, gender and social isolation in very late-onset schizophrenia like psychosis. AB - BACKGROUND: Data from two retrospective first contact studies suggest that the risk of developing very-late onset schizophrenia-like psychosis (SLP) may be raised in older migrant than British-born populations resident in the UK. OBJECTIVE: To investigate whether the relative excess of SLP observed amongst younger and male migrants in the above studies might have been explained by differences in the age-gender structures of migrant and British-born denominator populations. A secondary aim was to examine the associations of migrant group status and gender with markers of social isolation. METHOD: Eighty-six new referrals of SLP to the Mile End (1997-2003) and Maudsley (1995-2000) hospitals were identified from two retrospective case note studies. Local census data were used to estimate the denominator populations and to calculate rate ratios for migrant and British-born cases of SLP. Case notes were re-examined, to assess markers of social isolation in migrant and British-born patients. RESULTS: Migrant patients were more likely to be male (odds ratio = 4.8; CI(odds) = 1.8 13.2) than British-born patients. The ratio of first contact rates for migrant compared to British-born populations were highest amongst men. There was a lower mean age of onset of SLP in migrant than British-born patients (t = 4.30, 95% CI = 3.78-10.27), which was largely explained by a higher mean age of illness onset in British-born women. There were no differences between migrant and British-born patients with respect to markers of social isolation. Male patients were more likely to have never married than women (odds ratio = 0.28; 95% CI odds = 0.09 0.89). CONCLUSIONS: The age-gender structure of the background population is not sufficient to explain the socio-demographic differences between migrant and British-born patients with SLP. Male patients may be more socially isolated. PMID- 16250077 TI - Depression is the predominant factor contributing to morale as measured by the Philadelphia Geriatric Morale Scale in elderly Chinese aged 70 years and over. AB - OBJECTIVE: To examine factors contributing to the total Philadelphia Geriatric Morale Scale (PGMS) and its two subscales: reconciled ageing and unstrained affect. METHOD: The PGMS was administered to 759 community-living subjects aged 70 years and over. Information regarding socioeconomic status, health conditions, sensory impairment, physical symptoms, social support, activities of daily living as measured by the Barthel Index, life satisfaction, and the Geriatric Depression Score, was collected. Associations between these factors and PGMS and its subscale were examined using univariate analysis (Mann-Whitney; Kruskal-Wallis tests), and multivariate analysis using the classification and regression tree (CART) method. RESULTS: Gender, old age, physical, socioeconomic and social factors were significantly associated with PGMS. There was a strong correlation with GDS (r = 0.77, p < 0.001). In the CART analysis, for both subscales and the total score, GDS was the predominant factor contributing to the score. Other factors include self perception of health, enough expenses, overall satisfaction with life, gender, and constipation. DISCUSSION: The PGMS and GDS are closely related. In addition to the GDS, health perception, life satisfaction, and adequate finance were factors contributing to quality of life in elderly Hong Kong Chinese. PMID- 16250079 TI - Diagnostic accuracy of the original 30-item and shortened versions of the Geriatric Depression Scale in nursing home patients. AB - OBJECTIVE: To determine the diagnostic accuracy of the 30-item and shortened versions of the Geriatric Depression Scale (GDS) in diagnosing depression in older nursing home patients. METHOD: Three hundred and thirty-three older nursing home patients participated in a prospective cross-sectional study in the Netherlands. Sensitivity and specificity, positive and negative predictive values, and the area under the receiver operating curve (ROC) were assessed. Cronbach alphas were also calculated. Both major depression (MDD) and minor depression (MinD) according to the DSM-IV criteria, measured with the Schedules of Clinical Assessment in Neuropsychiatry (SCAN), were used as 'gold standard'. RESULTS: The cut-off point > or = 11 on the GDS-30 gave a sensitivity of 96.3% for MDD and 85.1% for MinD, with a specificity of 69.1%. The sensitivity of most of the shortened versions was sufficient, varying between 88.9% and 100% for MDD, and between 63.8% and 97.9% for MinD. With regard to the shortened versions, best sensitivity (96.3% and 78.7%) and specificity (69.5%) were found for the GDS-10 developed by D'Ath et al. (1994). The specificity rates for most of the shortened versions were found to be less satisfactory, varying between 18.9% and 74.1%. Sufficient internal consistency was found for the GDS-30, the GDS-15, the GDS-12 and the GDS-10, with Cronbach's alphas varying between 0.88 and 0.72. CONCLUSIONS: The GDS-30 was found to be a valid and reliable case-finding tool for both major and minor depression in nursing home patients with no cognitive impairment and in patients with mild to moderate cognitively impairment (MMSE > or = 15). The GDS-10 (D'Ath et al., 1994) appeared to be the best least time consuming alternative for the nursing home setting. PMID- 16250078 TI - MRI measures and progression of cognitive decline in nondemented elderly attending a memory clinic. AB - OBJECTIVE: To investigate whether MRI-based volumes of whole brain, medial temporal lobe and white matter hyperintensities (WMH) predict progression of cognitive decline in a sample of nondemented elderly. METHODS: Thirty-seven nondemented elderly attending a memory clinic and 28 elderly controls participated in this follow-up study. The average follow-up period was 1.8 years. Cognitive function was measured at baseline and follow-up with the Cambridge Cognitive Examination (CAMCOG). Baseline Magnetic Resonance Imaging (MRI) provided quantitative measures of whole brain, medial temporal lobe and WMH. Linear mixed models controlled for age and sex were used to assess the independent associations between MRI measures, baseline cognition, and annual decline in cognition. RESULTS: Medial temporal lobe volume was independently associated with baseline CAMCOG score (p < 0.01), whereas whole brain volume (p < 0.01) and WMH (p < 0.05) were associated with annual decline in CAMCOG score. CONCLUSIONS: These data suggest that regional damage to the medial temporal lobes underlies initial mild cognitive impairment, whereas more global brain changes, such as whole brain atrophy and WMH, contribute to further progression of cognitive decline. PMID- 16250080 TI - Deliberate self-harm in older people revisited. AB - BACKGROUND: Deliberate self harm (DSH) in later life is under researched and is believed to be related to both mental illness and suicide. AIMS: The aim of the study was to examine deliberate self-harm (DSH) in older people presenting to acute hospital services over three years. METHOD: This was a retrospective observational study. We reviewed 97 episodes of DSH involving 82 patients aged 65 and over referred to the Liaison Psychiatric Service of the Tees and North East Yorkshire NHS Trust South Locality from 2000 to 2002. RESULTS: There was a year on year increase in the number of older people presenting with DSH, especially in men. Twenty-one percent of older men had no discernible psychiatric diagnosis. There were a small number of people who repeated DSH within a year and males were as likely to be repeaters as females. Twenty-three percent of all patients saw a General Practitioner (GP) in the 7 days before the episode of DSH and this increased to 58% in the 4 weeks preceding the episode of DSH. More males (56%) than females (26%) who presented with DSH were married. The most common method of DSH (93%) was medication overdose of which 66% used prescribed medication. There was no difference in the methods used to self-harm between men or women. CONCLUSION: DSH in the elderly may start to mirror some of the characteristics seen in younger adults with DSH. While the numbers of DSH per year are small among the elderly compared to younger adults, the observations suggest an increase in DSH in men. Marriage may no longer be a protective factor in prevention of DSH among older men. Longer-term observational studies of DSH in older people are required to confirm these changing patterns. GPs may have an important role to play in prevention of DSH in later life. PMID- 16250081 TI - Prevalence and risk factors for depressive symptoms among community-dwelling elders in Taiwan. AB - BACKGROUND: Depression in the elderly has become a serious health care issue worldwide. However, no studies have determined the prevalence and risk factors for depressive symptoms among a representative sample of community-dwelling elders in Taiwan. OBJECTIVES: To determine the prevalence and risk factors for depressive symptoms among community-dwelling elders in Taiwan. METHODS: Stratified random sampling was used to recruit a representative sample of 1200 elderly participants from northern, middle, southern, and eastern regions of Taiwan. RESULTS: The prevalence of depressive symptoms was 27.5%. Logistic regression analysis demonstrated that having a respiratory disease, poor cognitive function, poor social support network, dissatisfaction with living situation, perception of poor health status, and perceived income inadequacy were significant predictors of depressive symptoms in this sample. CONCLUSIONS: The risk factors for depression identified in this study need to be considered when assessing the health of elderly persons. In addition, interventions to reduce depressive symptoms in elders should include strategies to change some of these modifiable risk factors. PMID- 16250082 TI - Prevalence of depression in older patients consulting their general practitioner in The Netherlands. AB - BACKGROUND: Little is known about the prevalence of depression in older patients consulting their general practitioner. AIM: To estimate the prevalence of major and minor depression in older GP attendees. DESIGN: Cross sectional two-stage screening design. SETTING: Seventeen GPs in urban and 41 GPs in rural settings. METHODS: Data of The West Friesland Study were used. Five thousand, six hundred and eight-six GP attendees of 55 years and older filled in the Geriatric Depression Scale-15 items as a screening instrument for depression (response 62%). Of those screened positive on the GDS-15, 846 (77.5%) were interviewed using the Primary Care Evaluation of Mental Disorders (PRIME-MD). A random sample (n = 102) scoring below the threshold of the GDS-15 was interviewed to estimate the proportion of false negatives. RESULTS: Major depression was prevalent in 13.7% and minor depression in 10.2% of the patients. Depressed patients were older (mean difference 0.7 years; p = 0.037), more often female (OR 1.3; p < 0.001) and lived more often in urban districts (OR 1.5; p < 0.001). Patients with major depression were younger (mean difference 1.9 years; p < 0.001) and more often female (OR 1.4; p = 0.026) than those with a minor depression. Only 22.9% of the patients with a major depression were treated with antidepressants. CONCLUSION: Depression in older GP attendees is a very common health problem. Further research should focus on identifying those groups of patients with high risk of persistence of depression. This could help to focus the limited resources available in general practice to those patients in whom treatment is most urgently needed. PMID- 16250083 TI - Letter regarding article by Patti et al, "Randomized trial of high loading dose of clopidogrel for reduction of periprocedural myocardial infarction in patients undergoing coronary intervention: results from the ARMYDA-2 (Antiplatelet therapy for Reduction of MYocardial Damage during Angioplasty) study". PMID- 16250084 TI - Effect of hypothermia on postmortem alterations in MAP2 immunostaining in the human hippocampus. AB - Ischemic neuronal injury induce degradation of microtubule-associated protein 2 (MAP2). In addition to ischemia, postmortem brains show alterations in MAP2 immunoreactivity in the hippocampus, suggesting that the factors inducing cytoskeletal disruption in postmortem brain are similar to those in ischemic brains. Hypothermia reduces the severity of ischemic injury including disruption of MAP2 in the hippocampus. However, whether hypothermia reduces postmortem changes of MAP2 was not clear. In this study, we evaluated the effect of hypothermia on postmortem degradation of MAP2 in the human hippocampus at various postmortem intervals using immunohistochemistry. In postmortem brains without hypothermia (the normothermic group), the locus of MAP2 immunoreactivity moved from the dendrites to the cell bodies prior to becoming undetectable with increasing postmortem interval, particularly in the CA1-subiculum region. On the other hand, the change in MAP2 immunoreactivity was remarkably attenuated in brains of death from cold (the hypothermic group). The present study demonstrated that MAP2 disruption is remarkable in the CA1-subiculum region of autopsied brains and that hypothermia reduces the postmortem change of MAP2, as observed in ischemic brain. Therefore, immunostaining of MAP2 in the hippocampus could be used to diagnose hypothermia. PMID- 16250085 TI - A trial of metoclopramide vs sumatriptan for the emergency department treatment of migraines. PMID- 16250086 TI - The Babinski sign: thumbs up or toes down? PMID- 16250087 TI - Medical malpractice--mixed eligibility and treatment decisions--preemption. Land v. CIGNA Healthcare of Florida. PMID- 16250088 TI - Statin treatment in first 24 hours after heart attack cuts mortality. PMID- 16250089 TI - Obesity may lead to loss of cognitive function. Those extra pounds you start adding in your forties raise the risk. PMID- 16250090 TI - AD reversed in mice--tau protein implicated. PMID- 16250091 TI - Therapy tripleheader extends life for non-small cell lung cancer patients. Concurrent radiation, chemotherapy and lobectomy increase survival for those with NSCLC. PMID- 16250092 TI - STD vaccine breakthrough. Cervarix would prevent human papilloma virus which can lead to cervical cancer; FDA approval anticipated. PMID- 16250093 TI - Freezing cancerous areas diminished bone cancer pain. PMID- 16250094 TI - Many at-risk patients not getting appropriate meds. poor and inconsistent compliance with anti-thrombotic therapy guidelines reported. PMID- 16250095 TI - Does sudden death run in your family? PMID- 16250096 TI - Lighten the load on your arthritic knees. Each pound you lose means four less pounds of pressure on your knee joint. PMID- 16250098 TI - Avoiding hip fractures: walking may not be enough. "Targeted exercises" might reduce the risk of hip fractures that comes with age. PMID- 16250097 TI - Slip-and-trip prevention. PMID- 16250099 TI - Yoga may help prevent weight gain, help weight loss. PMID- 16250100 TI - Ibuprofen before gum surgery doubles bleeding. NSAIDS are more effective in reducing pain and swelling when taken after surgery. PMID- 16250101 TI - I've heard that some exercises actually cause arthritis or make it worse. Is this true? PMID- 16250102 TI - Measurements of the quantum efficiency of discrimination in human scotopic vision. PMID- 16250103 TI - Expansion of triplex recognition codes by the use of novel bicyclic nucleoside derivatives (WNA). AB - Recently, we have developed new base analogs (WNA) and demonstrated that WNA-[see text];T with thymine and WNA-[see text];C with cytosine stabilize n on-natural antiparallel triplexes with a TA or CG interrupting site, respectively. However, limitations in recognizable sequences with the WNA-containing TFO were also found. The objective of this study is to search better WNA analogs for expansion of triplex recognition codes to general duplex sequences. In this study, we designed new WNA analogs by systematic modification of the aromatic part and the recognition part. The new WNA analogs with the benzene ring substituted with bromide or cyanide have determined for selective stabilization of triplexes at a TA interrupting site, and general formation of triplexes having a TA interrupting site has been achieved. PMID- 16250105 TI - Model modifications in covariance structure analysis: the problem of capitalization on chance. AB - In applications of covariance structure modeling in which an initial model does not fit sample data well, it has become common practice to modify that model to improve its fit. Because this process is data driven, it is inherently susceptible to capitalization on chance characteristics of the data, thus raising the question of whether model modifications generalize to other samples or to the population. This issue is discussed in detail and is explored empirically through sampling studies using 2 large sets of data. Results demonstrate that over repeated samples, model modifications may be very inconsistent and cross validation results may behave erratically. These findings lead to skepticism about generalizability of models resulting from data-driven modifications of an initial model. The use of alternative a priori models is recommended as a preferred strategy. PMID- 16250104 TI - [Effect of Chinese herbal medicine on patients with primary hepatic carcinoma in III stage during perioperational period: a report of 42 cases]. AB - OBJECTIVE: To investigate the synergetic effect of general therapy of Chinese herbal medicine with surgical operation. METHODS: Fourty-two patients in the integrative group were treated with jiedu xiaozheng yin for 7 days before operation, and fuzheng yiliu recipe after operation for 2 years, and 30 patients in the control group underwent operation alone. Their cellular immune function, survival rate and recurrence rate were compared and analyzed. RESULTS: The accumulative survival rate of 6-month, 12-month, 24-month and 36-month in the integrative group was 97.6%(41/42), 85.7%(36/42), 52.3% (22/42) and 45.5(17/42)% respectively and those in the control group 96.7% (29/30), 83.3% (25/30), 50.0% (15/30) and 30.0% (9/30), respectively, among them the 36-month survival rate was significantly different between the two groups (P < 0.05). The 24-month recurrence rate in the two groups was 54.8% and 80.0% respectively, the difference between the two was significant (P < 0.05). CONCLUSION: Administration of compound Chinese herbal medicine in peri-operational period has definite effect on primary hepatic carcinoma in III stage, it can improve patients' immune function, decrease the recurrence rate and increase the cumulative survival rate. PMID- 16250106 TI - Reflections on public bioethics: a view from the trenches. AB - For many reasons, and more than its predecessors, the President's Council on Bioethics has been the subject of much public attention and heated controversy. But little of that attention and controversy has been informed by knowledge of the Council's mission, its ways of working, and, most importantly, its actual work. This essay describes the Council's mission, discusses its public ways of working, and reviews the five major works produced during the Council's first term. In all its activities, the Council has sought to develop a richer bioethics, one that recognizes and tries to do justice to the deep issues of our humanity raised by the age of biotechnology. Believing that these issues are properly matters to be discussed and governed by the polity as a whole, the Council also has sought to contribute to a genuinely public or political bioethics, beyond the rule of "experts," scientific and bioethical. PMID- 16250107 TI - The Baroness's Comittee and the President's Council: ambition and alienation in public bioethics. AB - The President's Council on Bioethics has tried to make a distinctive contribution to the methodology of such public bodies in developing what it has styled a "richer bioethics." The Council's procedure contrasts with more modest methods of public bioethical deliberation employed by the United Kingdom's Warnock Committee. The practices of both bodies are held up against the backdrop of concerns about moral and political alienation, prompted by the limitations of moral reasoning and by moral dissent from state policy under even the most democratic of governments. Although the President's Council's rhetoric is often scrupulously conciliatory, recurring features of its argumentative practice are regrettably divisive. They order these things better in Britain. PMID- 16250108 TI - Promises and perils of public deliberation: contrasting two national bioethics commissions on embryonic stem cell research. AB - National bioethics commissions have struggled to develop ethically warranted methods for conducting their deliberations. The National Bioethics Advisory Commission in its report on stem cell research adopted an approach to public deliberation indebted to Rawls in that it sought common ground consistent with shared values and beliefs at the foundation of a well-ordered democracy. In contrast, although the research cloning and stem cell reports of the President's Council on Bioethics reveal that it broached two different methods of public deliberation--balancing goods and following an overarching moral principle--it adopted neither. Thereupon its primer mover, Leon Kass, influenced particularly by the approach of Leo Strauss, sought to develop a method of public deliberation guided by tradition and practical wisdom. When this failed, the Council fell back on a method that took account of shared fundamental values of a free democracy--a method remarkable akin to that employed by the National Bioethics Advisory Commission. Respect for diverse reasonable conceptions of the good in a democratic polity requires national bioethics commissions to seek and incorporate that which is valuable in opposing positions. PMID- 16250110 TI - Liquid chromatography problem solving and troubleshooting: Could you provide a little background about reversed-phase polymer columns and some hints where the columns would be most useful? PMID- 16250109 TI - Gas chromatography problem solving and troubleshooting: What is the deal with water-based samples on a capillary GC column? PMID- 16250111 TI - Deemed duration of marriage for widows/widowers and removal of restriction on benefits to children of military parents overseas. Final rules. AB - We are issuing these final rules to reflect in our regulations changes to the Social Security Act (the Act) made by two provisions in the Social Security Protection Act of 2004 (SSPA), enacted on March 2, 2004. One provision added a new situation in which the 9-month duration-of-marriage requirement for surviving spouses under title II of the Act is deemed to have been met. The other provision removed a restriction against payment of Supplemental Security Income (SSI) benefits, under title XVI of the Act, to certain blind or disabled children who were not eligible for SSI benefits the month before their military parents reported for duty outside the United States. PMID- 16250112 TI - TRICARE; sub-acute care program; uniform skilled nursing facility benefit; home health care benefit; adopting Medicare payment methods for skilled nursing facilities and home health care providers. Final rule. AB - This rule partially implements the TRICARE "sub-acute and long-term care program reform" enacted by Congress in the National Defense Authorization Act for Fiscal Year 2002, specifically: Establishment of "an effective, efficient, and integrated sub-acute care benefits program," with skilled nursing facility (SNF) and home health care benefits modeled after those of the Medicare program; adoption of Medicare payment methods for skilled nursing facility, home health care, and certain other institutional health care providers; adoption of Medicare rules on balance billing of beneficiaries, prohibiting it by institutional providers and limiting it by non-institutional providers; and change in the statutory exclusion of coverage for custodial and domiciliary care. PMID- 16250114 TI - Americans with Disabilities Act of 1990. Public Law No. 101-336. PMID- 16250113 TI - Medical devices; immunology and microbiology devices; classification of cystic fibrosis transmembrane conductance regulator gene mutation detection system. Final rule. AB - The Food and Drug Administration (FDA) is classifying the cystic fibrosis transmembrane conductance regulator (CFTR) gene mutation detection systems into class II (special controls). The special control that will apply to the device is the guidance document entitled "Class II Special Controls Guidance Document: CFTR Gene Mutation Detection Systems." The agency is classifying the device into class II (special controls) in order to provide a reasonable assurance of safety and effectiveness of the device. Elsewhere in this issue of the Federal Register, FDA is announcing the availability of the guidance document that will serve as the special control for the device. PMID- 16250115 TI - [Reviewing my article, "Mild cases of clouding of consciousness"]. PMID- 16250116 TI - [Legislative arguments on execution of mentally ill--special reference to the Constitution of the United States of America]. PMID- 16250117 TI - Respecting difference and moving beyond regulation: tasks for U.S. bioethics commissions in the twenty-first century. AB - This article focuses on two possible missions for a national bioethics commission. The first is handling differences of worldview, political orientation, and discipline. Recent work in political philosophy emphasizes regard for the dignity of difference manifested in "conversation" that seeks understanding rather than agreement. The President's Council on Bioethics gets a mixed review in this area. The second is experimenting with prophetic bioethics. "Prophetic bioethics" is a term coined by Daniel Callahan to describe an alternative to compromise-seeking "regulatory bioethics." It involves a critique of modern medicine. In the contemporary context, the areas of biotechnology and access to health care cry out for prophetic attention. The Council has addressed biotechnology; unfortunately, that experience suggests that the kind of prophecy that it practices poses risks to conversation. With regard to access issues, the article proposes an effort that unites themes of human dignity, solidarity, and limits in support of reform, while highlighting, rather than papering over, differences. PMID- 16250118 TI - Bioethics as politics: the limits of moral expertise. AB - The increasing reliance upon, and perhaps the growing public and professional skepticism about, the special expertise of bioethicists suggests the need to consider the limits of moral expertise. For all the talk about method in bioethics, we, bioethicists, are still rather far off the mark in understanding what we are doing, even when we may be going about what we are doing fairly well. Quite often, what is most fundamentally at stake, but equally often insufficiently acknowledged, are inherently political, essentially contested visions of the most compelling and attractive forms of life for individuals and social organization. The current situation in bioethics parallels similar debates in eighteenth-century jurisprudence, especially Jeremy Bentham's withering critique of the prevalent forms of judicial argument and his own, equally unsuccessful, attempt to develop a decision-making procedure in ethics that would operate on a plane above politics. The risk, both then and now, is that we will fail to appreciate the wide range of reasonable disagreement that will remain past the point of extended reflection and discussion. PMID- 16250119 TI - [Suggestions from the Japanese Society of Lawyers on the problems existing in the execution law in Japan]. PMID- 16250120 TI - [Role of psychiatrists in capital punishment]. PMID- 16250121 TI - [Drug adherence]. PMID- 16250122 TI - [Usefulness of PET in clinical trials of psychotropic drugs]. PMID- 16250123 TI - [Differential diagnosis of schizophrenia from cognitive disorders in AD/HD]. PMID- 16250124 TI - Intracervical application of synthetic hydrogel sealant for preterm prelabor rupture of membranes: a case report. AB - We report a case of prelabor rupture of membranes at 20 weeks' gestation where a hydrogel sealant was applied to the cervical canal in order to maintain an adequate liquor volume in the second trimester. The use of this agent for the purpose of creating plug has not been reported to date. PMID- 16250125 TI - Inspection focus will shift to commissioners by 2008. PMID- 16250126 TI - NICE appeals to DoH for faster referral of new drugs. PMID- 16250127 TI - Bid to boost independence in medical errors bill. PMID- 16250129 TI - On flying into trouble. PMID- 16250128 TI - Market. When F is for failure. PMID- 16250130 TI - The HSJ debate. Choosing Health is guiding the public to a healthy lifestyle. PMID- 16250131 TI - Data briefing. Day-case rates. PMID- 16250132 TI - Rebuilding Iraq. Healthcare under fire. PMID- 16250133 TI - Frontline management. PMID- 16250134 TI - On rural deprivation. PMID- 16250135 TI - Survey. Mixed blessings. PMID- 16250136 TI - Improvement. Five steps to tomorrow. PMID- 16250137 TI - Innovation. Hook up. PMID- 16250138 TI - Leadership. Reach for the top. PMID- 16250139 TI - Learning. Keep up the good work. PMID- 16250140 TI - Work relations. Difficult doctors. PMID- 16250142 TI - The case for healthcare. PMID- 16250141 TI - The battle over gay teens. PMID- 16250143 TI - Just don't do it! Are we teaching our kids way too much about sex? Or not nearly enough? PMID- 16250144 TI - Reshuffling the DRG deck. New cardiac DRGs praised for more equitable reimbursements while post-acute-transfer changes are generating some worries. AB - The CMS' changes to its DRGs, which took effect Oct. 1, could alter hospitals' bottom lines significantly. The addition of 16 DRGs makes a total of 526 classifications that will dictate payments and determine hospital services and patient mix. Tom Watson, left, a partner with accounting firm BKD, which specializes in healthcare reimbursement, says it's unlikely the changes will put any hospitals out of business, but they will feel a pinch. PMID- 16250145 TI - HHS looks for IT fast track. Approved products could hit market by next summer. PMID- 16250146 TI - Sidestepping Stark. HHS easing way for hospitals to help docs buy IT. PMID- 16250147 TI - Better quality, fewer patients. More plans report quality gains, but they cover fewer. PMID- 16250149 TI - HCA tries to think small. Company to reorganize hospitals into three groups. PMID- 16250148 TI - Ruling a win for Scruggs. Oregon judge grants class-action request. PMID- 16250150 TI - Catholic-Jewish deal in Ky. Joint venture in Louisville is called 'historic'. PMID- 16250151 TI - Connect the dots. Employers and insurers are behind the wheel on 'consumer-driven healthcare'. PMID- 16250152 TI - The business case for reform. Employers must demand needed change to create a sustainable system of care. PMID- 16250153 TI - Above-board? Governing bodies for GPOs come in many forms, but how they're formed can be less than transparent. PMID- 16250154 TI - Sizing up the market. Large IT companies paying little attention to physician practices. PMID- 16250155 TI - Partnership working in Scotland. PMID- 16250156 TI - The women the world forgot. PMID- 16250157 TI - Raising the profile of women's health. PMID- 16250158 TI - Internationally qualified midwives: developing a pathway to adaptation. AB - Midwives who have gained their qualifications abroad are a valuable asset to the NHS, but they require training in some areas to enable them to adapt to practice in the U.K. With a view to developing such a programme, research was undertaken into the communication skills and learning needs of internationally qualified midwives. PMID- 16250159 TI - Saxmundham one-stop health shop. PMID- 16250160 TI - The business of maternity services. PMID- 16250161 TI - Whose child? Ethics, practice and the law. PMID- 16250162 TI - Benefits of sure start caseload midwifery. AB - The Sure Start programme was devised to address inequalities in access to healthcare services and improve outcomes for vulnerable and disadvantaged groups. One such model is in Kennington and Nina Khazaezadeh explains how this service has offered new opportunities and enabled women to have more involvement in decisions regarding the care they receive. PMID- 16250163 TI - Leadership: the undervalued element? PMID- 16250164 TI - Benefits of Agenda for Change (AFC). PMID- 16250165 TI - The 'One mother one midwife' (OMOM) campaign. PMID- 16250166 TI - Anger over advanced midwife practitioner recruitment. PMID- 16250167 TI - Anger over advanced midwife practitioner recruitment. PMID- 16250168 TI - Growing old in the NHS. PMID- 16250169 TI - Air conditioning closely examined. AB - The intricacies of the provision and control of air conditioning in hospitals have been debated for decades. For this Health Estate Journal 'Then and now' feature, Mike Arrowsmith BSc (Hons) CEng FIMechE FIHEEM, technical editor, provides commentary on an article which was published in The Hospital Engineer, the journal of the Institution of Hospital Engineers, in July 1960. The text of this article, which was based on a paper read before the Glasgow Branch of the Institution, follows that of the commentary. PMID- 16250170 TI - Design and construct strategy explained. PMID- 16250171 TI - HSE role pivotal in legionella control. PMID- 16250172 TI - Show to be best yet. AB - This year's Healthcare Estates exhibition will be the largest ever, offering visitors even more opportunities to gather the latest information on everything from major hospital construction techniques to thermostatic mixing valves for water systems. Health Estate Journal previews some of the highlights. PMID- 16250173 TI - Making the case for recycled content. AB - Dr David Moon, head of procurement for the Waste & Resources Action Programme (WRAP), outlines how materials with higher recycled content can be economically sustainable, technically viable and easily available for construction projects in the healthcare sector. PMID- 16250174 TI - Copper comeback advocated. PMID- 16250175 TI - Pathology robotic systems excel. AB - The role of robotic sample processing in the modernisation of pathology services at Manchester Royal Infirmary is described by Peter Jones, centralised in vitro diagnostics market manager at Roche Diagnostics. Stemming from the modernisation are valuable pointers for managers of facilities and designers of healthcare premises. PMID- 16250176 TI - Global showcase for medical innovation. AB - Set against a backdrop of medical industry competitiveness and growth, the forthcoming MEDICA event will present ample opportunities for international exchange of ideas on a range of healthcare technology issues, says Horst Giesen, project leader, Messe Dusseldorf GmbH. PMID- 16250177 TI - Poor hygiene under new attack. AB - Attention-grabbing is a new campaign designed to provide assistance in the fight against healthcare acquired infections caused by MRSA. Health Estate Journal reports on the campaign, which is titled 'Wipe It Out' and is being run by the Royal College of Nursing and Kimberly-Clark. PMID- 16250178 TI - Coronary artery calcium and the risk of infarction: a method in search for a role? PMID- 16250179 TI - B-type natriuretic peptide: where are we? PMID- 16250180 TI - Acquired hemophilia A: a rare but important challenge for the internist. PMID- 16250181 TI - [Coronary calcium as an atherosclerosis marker]. AB - Non-invasive diagnostic techniques such as electron beam computed tomography and multislice spiral computed tomography are able to detect and quantify coronary calcifications. Several clinical studies have shown how the amount of coronary calcifications correlates to the coronary plaque burden. The detection of coronary calcium therefore provides a unique opportunity to identify and quantify coronary atherosclerosis in a subclinical stage. Measures of subclinical atherosclerosis may also help in recognizing factors related to atherosclerosis in asymptomatic populations. In addition, a significant proportion of subjects who develop premature clinical disease are not identified as being at high risk by current strategies. A scan negative for coronary calcium has a high negative predictive value indicating the absence of stenotic coronary artery disease. The aim of this review was to describe the potentials of coronary calcium detection and to summarize its clinical relevance. PMID- 16250182 TI - [Reproductive aspects of celiac disease]. AB - In the past, celiac disease (CD), or intolerance to gluten, was considered a rare disease of infancy characterized by chronic diarrhea with malabsorption and delayed growth. Besides the overt enteropathy, there are other clinic and subclinical forms which appear later in life. Target organs are not limited to the gut, but include liver, thyroid, skin and female and male reproductive systems. CD interference on reproduction is related to the multifactorial nature of the disease, whose pathological manifestations can be modulated, besides gluten, by different concurrent genetic and environmental factors. CD induces malabsorption with consequent deficiencies of micronutrients such as iron, folic acid and vitamin K, which are essential for organogenesis, and fat-soluble vitamins important for spermatogenesis. Regarding endocrine disorders, the deficiencies of specific trace elements on ovarian function could explain its involvement in the increased risk of female osteoporosis in CD patients. Affected males show a picture of tissue resistance to androgens; the increases of follicle stimulating hormone and prolactin, not associated with infertility, may indicate an imbalance at hypothalamus-pituitary level, with general effects on health. Since reproductive alterations are reversible, adoption of a gluten-free diet supported by early diagnosis is important. Therefore, the detection of early biomarkers, such as deficiencies of vitamins and/or iron and andrological or endocrinological dysfunctions, should trigger timely strategies for prevention and treatment. PMID- 16250183 TI - [Clinical nutritional outcome in patients recovering in a psychiatric setting from severe protein-energy malnutrition of anorexia nervosa]. AB - Protein energy malnutrition due to anorexia nervosa, either restrictive or bulimic, requires an integrated medical psychiatric intervention to be treated. The aim of this study was to evaluate the effectiveness of this integrated treatment in severely malnourished anorectic patients requiring to be hospitalized in Psychiatry Unit. Fifteen patients (14 females, 1 male, mean age 19.6 +/- 4.7 years, body mass index 14.0 +/- 1.9 kg/m2) 13 of whom affected by restrictive anorexia nervosa and 2 by bulimic anorexia nervosa, have been hospitalized in the Psychiatry Unit of the Federico II University Hospital, Naples from September 2000 to July 2003, always without requiring compulsory sanitary treatment. Hospitalization was due to failure of the outpatient treatment in all of them, complicated by uncontrolled weight loss in 7, hydroelectrolytic unbalance in 2, edema in 1 patient. All were hypotensive and 4 had marked bradycardia. Forced nutrition was never necessary. Enteral nutrition by nasogastric tube was prescribed in 4 patients, oral nutrition supplements with diet in 4 and only diet in the remaining 7. All patients received vitamin and mineral supplements, if necessary parenterally. A mild body weight increase and satisfactory normalization of biochemical parameters was obtained in all patients during hospitalization. Thereafter they were enrolled in an outpatient integrated medical/psychiatric protocol, including group therapy. Only in 1 case, a few months later, a second hospitalization was necessary. In conclusion, integrated medical psychiatric treatment represents an effective intervention also in severely malnourished anorectic patient requiring hospitalization. PMID- 16250185 TI - [Dercum's disease: a case report]. AB - A case of Dercum's disease in a 51-year-old obese woman with a history of brain vasculitis, and painful subcutaneous multiple lipomas is described. This disease, included in the category of rare diseases by the World Health Organization, first described in 1892 by Francis Xavier Dercum, is characterized by its prevalence among women, its familiarity, by the presence of multiple painful subcutaneous lipomas and its association with obesity, hypercholesterolemia and asthenia. The disease has to be differentiated from Madelung syndrome, the multiple familiar lipomatosis and Proteus' syndrome. The quality of life of the patients is often poor and since treatments so far have not been conclusive a better knowledge of the pathogenesis of the disease is desirable. PMID- 16250184 TI - [Variation in the plasma concentration of B-type natriuretic peptide in emergent paroxysmal atrial fibrillation, in acute pulmonary embolism, in acute coronary syndrome and in dilated cardiomyopathy]. AB - Our research is based on the critical evaluation of plasma concentration variation of B-type natriuretic peptide (BNP)--in emergency--in paroxysmal atrial fibrillation, acute pulmonary edema, acute coronary syndrome and dilated cardiomyopathy. The aim of our research was to assess if the BNP concentration variation may be useful in the diagnosis and therapy. Peptide synthesis takes place mainly in the ventricular myocardium. We selected 102 patients: 27 control subjects, and 75 admitted to the emergency and reception department for dyspnea and/or precordialgia and/or palpitations. At the beginning they were considered as one group only, and then they were divided into groups according to the diagnosis: 20 with paroxysmal atrial fibrillation with reversion to sinus rhythm in the first week; 20 with acute pulmonary edema; 22 with acute coronary syndrome without electrocardiographic ST-segment changes; 13 with compensated dilated cardiomyopathy. Our research assessed that the BNP activation and secretion are evident especially in patients with heart failure and remains at the high level until the administration of an effective therapy and then they reach a balance with values higher than the standards, while in the paroxysmal atrial fibrillation and in acute coronary syndrome they rise and come back to the standard levels or even at lower levels after the disease solution. For this reason, BNP reiterated measurements allow to assess treatment efficacy, even at home, and to optimize the therapy. The main limit of BNP diagnostic role is in the need of knowing in advance the specific values for each patient. The BNP concentration evaluation in the acute phase is necessary to differentiate patients with dyspnea due to heart failure from those with pulmonary pathologies, while the BNP assessment in the acute coronary syndrome predicted exitus or heart failure manifestations. PMID- 16250186 TI - [Acquired factor VIII hemophilia in a geriatric patient]. AB - Acquired hemophilia is a rare coagulopathy in adults, associated with bleeding complications. Although the etiology of this disorder remains obscure, an autoimmune mechanism produces the development of autoantibodies against factor VIII. About half of cases are associated with other conditions, mainly post partum, underlying cancer, autoimmune disease. An 81-year-old male was admitted to the hospital with extensive hematomas (neck, chest, arms and lower limbs). There was no family or personal history of congenital bleeding diathesis. He had chronic bronchitis and cerebrovascular disease; no drugs had been used during the month prior to noted symptoms. Laboratory parameters revealed: hemoglobin 10.9 g%, normal platelet count and white blood cells, prolonged activated partial thromboplastin time (98 s), with normal prothrombin time and fibrinogen concentration. An activated partial thromboplastin time mixing study did not show any correction, suggesting a coagulation inhibitor. Lupus anticoagulant and anticardiolipin antibodies were negative. Biochemical, immunological tests and tumor markers were normal. Thoracic and abdominal computed tomographic scan did not reveal pathological images or hematomas. Analysis of clotting factors revealed decreased factor VIII (< 2%) and elevated factor VIII inhibitor (55 Bethesda units). Idiopathic acquired hemophilia diagnosis was made. Red blood cell transfusion and human factor VIII (2000 U/day for 7 days) infusion were initiated, intravenously with methylprednisolone. A progressive improvement in clinical conditions and laboratory parameters was observed. After 18 days the patient was discharged and treated with prednisone. At follow-up control the clinical conditions and laboratory parameters were normal. PMID- 16250187 TI - Association between immune thrombocytopenic purpura and chronic lymphocytic leukemia in a patient carrier of anti-hepatitis C virus antibodies. AB - Immune thrombocytopenic purpura (ITP) occurs in 2-3% of chronic lymphocytic leukemia (CLL) patients, whereas autoimmune thrombocytopenia is very rare before the diagnosis of lymphoma. A 67-year-old patient, was admitted to our Department because of purpura on his inferior limbs. Family history revealed arterial hypertension, a previous presence of hepatitis C virus (HCV) antibodies, with no sign of liver damage. Physical examination showed purpura of inferior limbs. Laboratory analysis revealed: marked thrombocytopenia (platelet count 5000/microL); hypogammaglobulinemia (9%, immunoglobulin-IgG 634 mg/dL); presence of HCV antibody (negative HCV-RNA); low-titer anti-nuclear antibody and anti smooth muscle antibody (1:80); positive cryoglobulin (polycolonal, IgG-IgM, cryocrit 0.5%). Abdomen ultrasound revealed a mild liver steatosis and bone marrow aspirate megakaryocytic hyperplasia. Platelet kinetics study showed a markedly reduced platelet half-life (<1 day) with evident splenic uptake. The patient was treated with steroids, intravenous Ig and immunosuppressive agent (cyclophosphamide) with only temporary effect; a splenectomy was therefore performed with a subsequent durable increase in the platelet count. Two years later, the patient underwent a prostatectomy for prostate cancer and within the pelvic nodal screening the histological examination unexpectedly revealed features of B-cell non-Hodgkin's lymphoma, type CCL/small lymphocytic lymphoma; a bone marrow aspirate showed a monotypic CD5+, CD19+, CD23+ B-cell proliferation confirming the diagnosis of CLL. Six months later, a computed tomography scan revealed multiple pathological node enlargements (1.5-3 cm), compatible with a malignant lymphoma. The marked thrombocytopenia may have been an early expression of the lymphoproliferative disease. Otherwise, the association between CLL and ITP might reflect the underlying role of HCV infection causing an immune dysregulation responsible for both pathologies. PMID- 16250188 TI - [Ertapenem. A new carbapenem in a single dose for community acquired infection]. PMID- 16250189 TI - Effects of external rhythmical cueing on gait in patients with Parkinson's disease: a systematic review. AB - OBJECTIVE: To critically review studies evaluating the effects of external rhythmical cueing on gait in patients with Parkinson's disease. METHODS: Articles published from 1966 to January 2005 were searched by two physiotherapists in MEDLINE, PiCarta, PEDRo, Cochrane, DocOnline, CINAHL and SUMSEARCH. To be included, articles had to investigate the effects of external rhythmical cueing (i.e., auditory, visual or tactile cueing) on gait parameters in patients with idiopathic Parkinson's disease. Both controlled and noncontrolled studies were included. Based on the type of design and methodological quality a meta-analysis or best-evidence synthesis was applied. RESULTS: Twenty-four studies (total number of patients = 626) out of the 159 screened studies were evaluated in this systematic review. Two out of 24 were randomized controlled trails (RCT), both of high methodological quality. One RCT did not focus specifically on external rhythmical cueing of individual patients with Parkinson's disease, but on group exercises in general, including walking with cues. All other studies were pre experimental studies. Best-evidence synthesis showed strong evidence for improving walking speed with the help of auditory cues. Insufficient evidence was found for the effectiveness of visual and somatosensory cueing. CONCLUSION: Only one high-quality study, specifically focused on the effects of auditory rhythmical cueing, suggesting that the walking speed of patients with Parkinson's disease can be positively influenced. However, it is unclear whether positive effects identified in the laboratory can be generalized to improved activities of daily living (ADLs) and reduced frequency of falls in the community. In addition, the sustainability of a cueing training programme remains uncertain. PMID- 16250190 TI - Impairment-oriented training or Bobath therapy for severe arm paresis after stroke: a single-blind, multicentre randomized controlled trial. AB - OBJECTIVE: To study the effects of augmented exercise therapy time for arm rehabilitation as either Bobath therapy or the impairment-oriented training (Arm BASIS training) in stroke patients with arm severe paresis. DESIGN: Single blind, multicentre randomized control trial. SETTING: Three inpatient neurorehabilitation centres. SUBJECTS: Sixty-two anterior circulation ischaemic stroke patients. INTERVENTIONS: Random assignment to three group: (A) no augmented exercise therapy time, (B) augmented exercise therapy time as Bobath therapy and (C) augmented exercise therapy time as Arm BASIS training. MAIN MEASURES: MAIN OUTCOME MEASURE: Fugl-Meyer arm motor score. Secondary measure: Action Research Arm Test (ARA). Ancillary measures: Fugl-Meyer arm sensation and joint motion/pain scores and the Ashworth Scale (elbow flexors). RESULTS: An overall effect of augmented exercise therapy time on Fugl-Meyer scores after four weeks was not corroborated (mean and 95% confidence interval (CI) of change scores: no augmented exercise therapy time (n=20) 8.8, 5.2-12.3; augmented exercise therapy time (n=40) 9.9, 6.8-13.9; p = 0.2657). The group who received the augmented exercise therapy time as Arm BASIS training (n=20) had, however, higher gains than the group receiving the augmented exercise therapy time as Bobath therapy (n=20) (mean and 95% CI of change scores: Bobath 7.2, 2.6-11.8; BASIS 12.6, 8.4-16.8; p = 0.0432). Passive joint motion/pain deteriorated less in the group who received BASIS training (mean and 95% CI of change scores: Bobath 3.2, -5.2 to -1.1; BASIS 0.1, -1.8-2.0; p = 0.0090). ARA, Fugl-Meyer arm sensation, and Ashworth Scale scores were not differentially affected. CONCLUSIONS: The augmented exercise therapy time as Arm BASIS training enhanced selective motor control. Type of training was more relevant for recovery of motor control than therapeutic time spent. PMID- 16250191 TI - Intensification of the transition between inpatient neurological rehabilitation and home care of stroke patients. Controlled clinical trial with follow-up assessment six months after discharge. AB - OBJECTIVE: An intensified transition concept between neurological inpatient rehabilitation and home care was investigated for effects on the functional status of stroke patients and the physical and emotional health of their carers. DESIGN: Controlled clinical trial allocating patients to intervention group (intensified transition on ward II) or control group (standard transition on ward I); patients were allocated to whichever ward had a vacancy. Follow-up assessment was carried out six months after discharge. SUBJECTS: Seventy-one patients and their family carers were included, of which nine cases dropped out. Therefore 62 stroke patients with persisting disability and their family carers were available for assessment at follow-up--33 patients in the intervention group, 29 patients in the control group. INTERVENTION: The intensified transition concept consisted of therapeutic weekend care, bedside teaching and structured information for relatives during the second phase of the rehabilitation. MAIN MEASURES: Patients were assessed with the Barthel Index, Functional Independence Measure, Ashworth Spastic Scale, Frenchay Arm Test, and Timed Up and Go Test. The carers completed SF-36, and were assessed using the Giessen Symptom List, Depression Scale and Burden Scale for Family Caregivers. RESULTS: The intensified transition did not lead to significant change in the functional status of the patients or in the physical and emotional health of the family carers. Within the first four weeks after discharge, the patients in the intervention group had fewer new illnesses. In the observation period the use of outpatient care services was more frequent in the intervention group than in the control group. CONCLUSION: Even though there are few differences of moderate intensity between the two groups the intensified transition programme does not affect either the functional status of the stroke patients or the health of the carers. PMID- 16250192 TI - Home-based electromyography-triggered stimulation in chronic stroke. AB - OBJECTIVES: (1) To determine the feasibility of a home-based electromyography triggered neuromuscular stimulation (ETMS) programme; and (2) to determine ETMS efficacy in increasing affected wrist extension and reducing affected arm impairment. DESIGN: Randomized, controlled, pre-post, cross-over design. SETTING: Outpatient rehabilitation hospital. PATIENTS: Twelve chronic stroke patients with palpable muscle contraction in their affected wrist extensors but no movement (7 males; mean age = 59.75 years, age range 44-75 years; mean time since stroke = 52.75 months, range 13-131 months). INTERVENTION: Subjects were randomly assigned to receive either: (a) ETMS use twice every weekday in 35-min increments during an eight-week period followed by an eight-week home exercise programme (ETMS/home exercise programme) (n=8); or (b) an eight-week home exercise programme followed by use of ETMS twice every weekday in 35-min increments during an eight-week period (home exercise programme) (n=4). MAIN OUTCOME MEASURES: The Fugl-Meyer, Action Research Arm Test and goniometry. RESULTS: After home exercise programme participation, subjects showed nominal or no changes on any of the outcome measures. After ETMS, patients showed modest impairment reductions, as shown by the Fugl-Meyer, and no Action Research Arm Test changes. However, both groups showed a 21 degree increase in active affected wrist extension after ETMS use. CONCLUSION: ETMS use is feasible in the home environment. Neither participation in a traditional home exercise programme nor ETMS use conveyed changes on the Fugl-Meyer or Action Research Arm Test. However, ETMS use increased active affected limb extension. This new movement may provide a potential pathway for subjects to participate in other interventions, such as modified constraint induced therapy. PMID- 16250193 TI - The intra-rater reliability of the balance performance monitor when measuring sitting symmetry and weight-shift activity after stroke in a community setting. AB - OBJECTIVE: To examine the intra-rater reliability of sitting symmetry and weight shift activity measurements in poststroke adults. DESIGN: An intra-rater reliability study. SETTING: A community setting. SUBJECTS: Adult stroke survivors attending stroke support groups within the community of Nottingham (U.K.). MAIN MEASURES: The Balance Performance Monitor used to measure sitting symmetry and weight-shift activity. Intraclass correlation coefficients (ICCs) and their 95% confidence intervals (95% CI) were calculated. The Bland Altman method for assessing agreement is also presented. RESULTS: We tested 49 participants (median age 73 years; interquartile range 68-81 years). Between-test reliability for sitting symmetry was high: ICC (1,1) = 0.93 (95% CI 0.87 < or = ICC < or = 0.96). The mean difference between the measures (d) was -0.08 (95% CI -0.48 < or = d < or = 0.31); the standard deviation of the differences (SDdiff) was 1.383. The coefficient of repeatability was 2.76; the 95% limits of agreement were -2.850 and 2.682. Between-test reliability for weight-shift activity was also high: ICC (1,1) = 0.86 (95% CI 0.77 < or = ICC < or = 0.92). Bland-Altman d = -0.08 (95% CI -0.19 < or = d < or = 0.35), SDdiff = 0.936. The coefficient of repeatability was 1.87; the 95% limits of agreement were -1.792 and 1.952. CONCLUSIONS: The 95% CI for d for both parameters crossed zero, indicating that between-test bias is unlikely. Sitting symmetry and weight-shift activity measures demonstrated acceptable levels of reliability. PMID- 16250194 TI - Reliability of the Modified Tardieu Scale and the Modified Ashworth Scale in adult patients with severe brain injury: a comparison study. AB - OBJECTIVE: To assess and to compare the reliability of the Modified Tardieu Scale with the Modified Ashworth Scale in patients with severe brain injury and impaired consciousness. DESIGN: Cross-sectional observational comparison study. SETTING: An early rehabilitation centre for adults with neurological disorders. SUBJECTS: Thirty patients with impaired consciousness due to severe cerebral damage of various aetiologies. MEASUREMENT PROTOCOL: Four experienced physical therapists rated each patient in a randomized order once daily for two consecutive days. Shoulder, elbow, wrist, hip, knee and ankle spasticity were assessed by the use of Modified Tardieu Scale and Modified Ashworth Scale data collection procedures. MAIN OUTCOME MEASURES: Test-retest and inter-rater reliability (kappa = kappa value) of the Modified Tardieu Scale and the Modified Ashworth Scale. RESULTS: The test-retest reliability of the Modified Ashworth Scale was moderate to good (kappa = 0.47-0.62) and of the Modified Tardieu Scale moderate to very good (kappa = 0.52-0.87). Test-retest reliability was significantly higher within the Modified Tardieu Scale in comparison with the Modified Ashworth Scale (Z > 1.96; p < 0.05) except for shoulder extensor and internal rotator muscles (Z < 1.96; p > 0.05). Although inter-rater reliability of both scales was poor to moderate (Modified Ashworth Scale: kappa = 0.16-0.42; Modified Tardieu Scale: kappa = 0.29-0.53), significantly higher K-values were revealed with the Modified Tardieu Scale for all tested muscle groups (Z > 1.96; p < 0.05) except for wrist extensors (Z < 1.96; p > 0.05). CONCLUSION: In patients with severe brain injury and impaired consciousness the Modified Tardieu Scale provides higher test retest and inter-rater reliability compared with the Modified Ashworth Scale and may therefore be a more valid spasticity scale in adults. PMID- 16250195 TI - Electromyography characterization of stretch responses in hemiparetic stroke patients and their relationship with the Modified Ashworth scale. AB - OBJECTIVES: To determine the validity of the Modified Ashworth Scale as a measure of spasticity by determining its relationship to surface electromyography activity and contracture. DESIGN: A controlled study of hemiparetic stroke patients with spasticity. SETTING: A physiotherapy department in a secondary care hospital. SUBJECTS: Thirty-one stroke patients and 20 healthy volunteers. MAIN MEASURES: The resistance to passive movement around the knee and ankle of the affected and unaffected legs was rated using the Modified Ashworth Scale. Passive range of movement was measured with a goniometer. Surface electromyography recordings of four lower limb muscles were taken during passive stretches of the knee and ankle. RESULTS: Hemiparetic patients produced surface electromyography responses to stretch that were of greater amplitude (unaffected limbs: mean = 25.82 mV (43.85), affected limbs: mean = 24.77 mV (35.46)) than those of healthy volunteers (mean = 15.85 (29.96)). The affected muscles of hemiparetic patients were more likely to produce surface electromyography responses to stretch of a sustained duration (45% of cases) compared with unaffected limbs (24% of cases) and those of healthy volunteers (16% of cases). The Modified Ashworth Scale showed a positive correlation with the magnitude (p < 0.05) and duration (p < 0.001) of the surface electromyography response. High scores on the Modified Ashworth Scale were associated with contracture (p < 0.001). Contracted muscles produced significantly greater surface electromyography reflex responses compared with noncontracted muscles (p < 0.05). CONCLUSION: The Modified Ashworth Scale reflects spasticity in terms of surface electromyography stretch responses produced by passive movement, but the relationship of spasticity to contracture remains unclear. PMID- 16250196 TI - Reliability of the positive and negative affect schedule (PANAS) in medical rehabilitation. AB - OBJECTIVE: To examine the reliability of the Positive and Negative Affect Schedule (PANAS) in patients 90 and 100 days after discharge from inpatient medical rehabilitation. DESIGN: A prospective study. SUBJECTS: A convenience sample of 31 medical rehabilitation patients. MAIN MEASURES: The PANAS. RESULTS: The average age was 74 years, 80.7% were female and the average length of stay was 13 days. The test-retest intraclass correlation (ICC) values for the positive and negative affect scales (at the 90 and 100 day follow-up assessment) were 0.79 and 0.93, respectively. CONCLUSIONS: Our results show that the PANAS has excellent reliability among a sample of patients who received inpatient medical rehabilitation. PMID- 16250197 TI - The Stroke Unit Discharge Guideline, a prognostic framework for the discharge outcome from the hospital stroke unit. A prospective cohort study. AB - OBJECTIVE: To investigate which factors during the subacute phase post stroke have predictive value for the discharge outcome from the hospital stroke unit. METHODS: In a prospective cohort of 338 patients admitted to a hospital stroke unit 26 potentially prognostic factors, arranged in clinical and social subdomains, were scored and analysed by binary logistic regression analysis. The outcome of the research consisted of the various discharge destinations. RESULTS: The overall predictive value of the discharge model is high (91%). Factors predictive of a poor discharge outcome are a low Barthel Index score (odds ratio (OR) 0.78 per point increase; p < 0.001), a poor sitting balance (OR 5.96; p < 0.001), a depression (OR 7.23; p < 0.001), poststroke cognitive disability (OR 3.51; p = 0.007) and older age (OR 1.05 per point increase; p = 0.008). If present, a personality disorder, premorbid cognitive disability and premorbid functional disability all show a tendency towards poor discharge outcome, but these factors did not reach statistical significance in this study, possibly due to their low prevalence. Readiness of the family circle to provide support was only significant in the univariate analysis. CONCLUSIONS: Somatic, biological and psychological factors predict the discharge outcome. Functional and cognitive factors play a decisive role in the future ability to live independently after a stroke. The prognostic importance of social factors could not be demonstrated. Urinary incontinence did not emerge as a prognostic factor. This is in contrast to scientific findings till now, but in accordance with clinical experience. PMID- 16250198 TI - A new method for predicting functional recovery of stroke patients with hemiplegia: logarithmic modelling. AB - OBJECTIVE: To examine the validity and applicability of logarithmic modelling for predicting functional recovery of stroke patients with hemiplegia. DESIGN: Longitudinal postal survey. SUBJECTS: Stroke patients with hemiplegia staying in a long-term rehabilitation facility, who had been referred from acute medical service 30-60 days after onset. METHODS: Functional Independence Measure (FIM) scores were periodically assessed during hospitalization. For each individual, a logarithmic formula that was scaled by an interval increase in FIM scores during the initial 2-6 weeks was used for predicting functional recovery. RESULTS: For the study, we recruited 18 patients who showed a wide variety of disability levels on admission (FIM scores 25-107). For each patient, the predicted FIM scores derived from the logarithmic formula matched the actual change in FIM scores. The changes predicted the recovery of motor rather than cognitive functions. Regression analysis showed a close fit between logarithmic modelling and actual FIM scores (across-subject R2 = 0.945). CONCLUSIONS: Provided with two initial time-point samplings, logarithmic modelling allows accurate prediction of functional recovery for individuals. Because the modelling is mathematically simple, it can be widely applied in daily clinical practice. PMID- 16250199 TI - Optimizing rehabilitation for adults with visual impairment: attention to life goals and their links to well-being. AB - OBJECTIVE: To examine the importance of different life goals among working-age adults with vision impairment, the extent to which vision impairment interfered with goals, and how rehabilitation addressed these life goals. DESIGN: Cross sectional descriptive study. SETTING: Vision rehabilitation agency. SUBJECTS: Working-age adults with visual impairment. METHOD: Telephone interviews using structured and open-ended assessments of life goal importance, goal interference due to vision loss, the role of goals in rehabilitation, and indicators of well being. RESULTS: Eighty-six people participated. The life domains most frequently rated as extremely important were finances (60), residential and domestic arrangements (55), family (51), partner (48), and personal care (48). The extent to which vision loss interferes with these life goals was extreme for finances (46), residential and domestic arrangements (36), partner (27), family (25), and personal care (14). A life goal was addressed in rehabilitation: residential/domestic arrangements (41), work (39), finances (31), leisure/hobbies (28), personal care (24), family (17), partner (16), friends (10), and religion/life philosophy (8). Reports of a life goal being addressed in ineffective ways emerged for functional life goals, but not for relationship and religion/life philosophy goals. Goal interference was significantly correlated with well-being indicators across life domains. In contrast only partner and family relationship importance were significantly linked with well-being indicators. CONCLUSIONS: Relationship-related goals were a top priority for this study population, but functional compared to relationship goals were more commonly addressed in vision rehabilitation services. PMID- 16250201 TI - New bill asks physicians to say 'l'm sorry' to avoid a med mal suit. PMID- 16250200 TI - Inferring personality traits of clients with Parkinson's disease from their descriptions of favourite activities. AB - OBJECTIVE: To explore the extent to which practitioners effectively use cues from clients' verbal descriptions of their favourite activities to form accurate impressions of the personality of clients with Parkinson's disease. PARTICIPANTS: Ninety-nine practitioners from disciplines of occupational, physical and speech therapy, nursing or medicine. PROCEDURE: Six men and six women with Parkinson's disease completed a self-report measure of personality and were individually interviewed regarding their favourite activities. The practitioners viewed 2-min segments of those videotaped interviews and provided judgements of the clients' personality. MEASURE: The NEO-Five Factor Inventory and a coding scheme to describe characteristics of clients' favourite activities. ANALYSIS: Clients' self-reported personality was correlated with the activity characteristics to identify the degree to which each characteristic was a cue of personality. Practitioners' judgements of personality were correlated with the activity characteristics to identify how heavily the practitioners weighted each cue. These two sets of weightings were compared using Pearson's correlations to determine whether practitioners used an appropriate cue strategy related to the activity descriptions. RESULTS: Practitioners appropriately used the personality cues found in the clients' favourite activity descriptions to assess the traits of Openness to Experience, Agreeableness and Conscientiousness (r = 0.66, r = 0.60, and r = 0.55, respectively, all p < or = 0.02). Practitioners appeared to use less effective cue strategies for the traits of Neuroticism and Extraversion. CONCLUSION: Clients with Parkinson's disease appear to express their personality in their descriptions of favourite activities, and practitioners appear to make use of these expressive verbal cues effectively for some aspects of personality. PMID- 16250202 TI - Managed care payments, drug prices ripe for savings efforts. PMID- 16250203 TI - United States has a 'patchwork system' of health coverage. PMID- 16250204 TI - Clinical and pathological features of pachyonychia congenita. AB - Pachyonychia congenita (PC) is a rare genodermatosis affecting the nails, skin, oral mucosae, larynx, hair, and teeth. Pathogenic mutations in keratins K6a or K16 are associated with the PC-1 phenotype whereas K6b and K17 mutations are associated with the PC-2 phenotype. Analysis of clinical, pathological, and genetic data from the literature and two research registries reveal that >97% of PC cases exhibit fingernail and toenail thickening, and painful plantar keratoderma. Prospective evaluation of 57 PC patients from 41 families revealed variable clinical findings: hyperhidrosis (79%), oral leukokeratosis (75%), follicular keratosis (65%), palmar keratoderma (60%), cutaneous cysts (35%), hoarseness or laryngeal involvement (16%), coarse or twisted hair (26%), early primary tooth loss (14%), and presence of natal or prenatal teeth (2%). Stratification of these data by keratin mutation confirmed the increased incidence of cyst formation and natal teeth among PC-2 patients, although cysts were more commonly seen in PC-1 than previously reported (25%-33%). Previously unreported clinical features of PC include development of painful oral and nipple lesions during breastfeeding, copious production of waxy material in ears, and inability to walk without an ambulatory aid (50%). Possible pathogenic mechanisms are discussed with respect to the clinicopathologic and genetic correlations observed. PMID- 16250205 TI - Treatment of pachyonychia congenita. AB - There are currently no specific treatments for pachyonychia congenita (PC). Available treatments generally are directed at specific manifestations of the disorder, and an effective treatment plan must recognize that different patients are more or less troubled by different manifestations of the disease. Treatment for all aspects of PC has been less than completely satisfactory. Very few studies have compared different approaches to treatment, and fewer still have given longitudinal follow-up of efficacy and patient acceptance. This review is essentially a compilation of anecdotes. It was collected from physicians' reports in the literature, from direct communication with physicians currently following patients with PC and from patients who answered a questionnaire on the Pachyonychia Congenita Project web page (http://www.pachyonychia.org/Registry.html). PMID- 16250206 TI - The genetic basis of pachyonychia congenita. AB - In 1994, the molecular basis of pachyonychia congenita (PC) was elucidated. Four keratin genes are associated with the major subtypes of PC: K6a or K16 defects cause PC-1; and mutations in K6b or K17 cause PC-2. Mutations in keratins, the epithelial-specific intermediate filament proteins, result in aberrant cytoskeletal networks which present clinically as a variety of epithelial fragility phenotypes. To date, mutations in 20 keratin genes are associated with human disorders. Here, we review the genetic basis of PC and report 30 new PC mutations. Of these, 25 mutations were found in PC-1 families and five mutations were identified in PC-2 kindreds. All mutations identified were heterozygous amino acid substitutions or small in-frame deletion mutations with the exception of an unusual mutation in a sporadic case of PC-1. The latter carried a 117 bp duplication resulting in a 39 amino acid insertion in the 2B domain of K6a. Also of note was mutation L388P in K17, which is the first genetic defect identified in the helix termination motif of this protein. Understanding the genetic basis of these disorders allows better counseling for patients and paves the way for therapy development. PMID- 16250207 TI - Insights into genotype-phenotype correlation in pachyonychia congenita from the human intermediate filament mutation database. AB - Keratins are the intermediate filament proteins specifically expressed by epithelial cells. The Human Genome Project has uncovered a total of 54 functional keratin genes that are differentially expressed in specific epithelial structures of the body, many of which involve the epidermis and its appendages. Pachyonychia congenita (PC) is a group of autosomal dominant genodermatoses affecting the nails, thick skin and other ectodermal structures, according to specific sub type. The major clinical variants of the disorder (PC-1 and PC-2) are known to be caused by dominant-negative mutations in one of four differentiation-specific keratins: K6a, K6b, K16, and K17. A total of 20 human keratin genes are currently linked to single-gene disorders or are predisposing factors in complex traits. In addition, a further six intermediate filament genes have been linked to other non epithelial genetic disorders. We have established a comprehensive mutation database that catalogs all published independent occurrences of intermediate filament mutations (http://www.interfil.org), with details of phenotypes, published papers, patient support groups and other information. Here, we review the genotype-phenotype trends emerging from the spectrum of mutations in these genes and apply these correlations to make predictions about PC phenotypes based on the site of mutation and keratin pair involved. PMID- 16250208 TI - Mouse models in preclinical studies for pachyonychia congenita. AB - The similarities between the human and mouse genomes often allow researchers to make accurate predictions about the roles of their human counterparts. Because of the similar physiology between these two mammals, mice are used extensively in the laboratory to investigate the mechanisms of human diseases. Furthermore, mice provide us with the option of testing the toxicity of drugs and the safety of therapeutic approaches prior to human application. Here, we review the existing mouse models involving the keratin genes (K6a, K6b, K16, and K17) that cause the human genetic disorder pachyonychia congenita (PC). We also suggest methods to more accurately model this autosomal dominant skin condition in the mouse in order to better understand the pathophysiological processes underlying PC and importantly, provide a test-bed for testing emerging therapies in vivo. PMID- 16250209 TI - Gene therapy for autosomal dominant disorders of keratin. AB - Dominant mutations that interfere with the assembly of keratin filaments cause painful and disfiguring epidermal diseases like pachyonychia congenita and epidermolysis bullosa simplex. Genetic therapies for such diseases must either suppress the production of the toxic proteins or correct the genetic defect in the chromosome. Because epidermal skin cells may be genetically modified in tissue culture or in situ, gene correction is a legitimate goal for keratin diseases. In addition, recent innovations, such as RNA interference in animals, make an RNA knockdown approach plausible in the near future. Although agents of RNA reduction (small interfering RNA, ribozymes, triplex oligonucleotides, or antisense DNA) can be delivered as nucleotides, the impermeability of the skin to large charged molecules presents a serious impediment. Using viral vectors to deliver genes for selective inhibitors of gene expression presents an attractive alternative for long-term treatment of genetic disease in the skin. PMID- 16250210 TI - Challenges in developing therapies for rare diseases including pachyonychia congenita. AB - The ability to attract sufficient resources to effectively develop therapeutics for rare diseases is a daunting task. This review summarizes existing resources for rare diseases and discusses some of the challenges and strategies associated with developing therapies for small patient populations with an emphasis on pachyonychia congenita. PMID- 16250211 TI - Outcome of treatment of extraocular muscle cysticercosis. PMID- 16250212 TI - Reduced inflammatory response after laser photocoagulation compared with cryoablation for threshold retinopathy of prematurity. PMID- 16250213 TI - Management of dissociated vertical deviation. Interview by Leonard B Nelson. PMID- 16250214 TI - What's your diagnosis? Oculocerebrorenal syndrome of Lowe. PMID- 16250215 TI - Hemihang-back recession. PMID- 16250216 TI - Ahmed valve surgery for refractory pediatric glaucoma: a report of 52 eyes. AB - PURPOSE: To describe the results of Ahmed valve surgery for refractory pediatric glaucoma. PATIENTS AND METHODS: We performed a retrospective review of patients younger than 18 years of age who had Ahmed valve surgery from November 1994 to January 2003. Success was defined as a reduction of the intraocular pressure (IOP) to 22 mm Hg or lower with or without medications at the last two follow-up visits, no additional glaucoma surgery, and no visually significant complications. RESULTS: There were 52 eyes of 41 patients. The two most common diagnoses were congenital glaucoma (38.5%) and aphakic glaucoma (36.5%). The mean number of glaucoma surgeries before Ahmed valve implantation was 1.7 +/- 1.7. The mean age at the time of implantation was 4.9 +/- 6.5 years. The mean IOP decreased from 38.1 +/- 6.4 mm Hg to 20.7 +/- 8.2 mm Hg at last follow-up. The final visual acuity was improved or within one Snellen line in 81.5% of the eyes. Cumulative probabilities of success were 85.1%, 63.2%, 51.7%, and 41.8% at 1, 2, 3, and 4 years, respectively. The mean postoperative follow-up period or time to failure for all patients was 2.2 +/- 1.8 years (range, 3 months to 7.5 years). CONCLUSIONS: The Ahmed valve was found to be useful in the management of refractory pediatric glaucoma. Although our success rates were similar to those observed in adults, the rates of certain post-operative complications were different. PMID- 16250217 TI - Effect of posterior tenectomy of the superior oblique on objective and subjective torsion in cases of superior oblique overaction. AB - PURPOSE: To evaluate the effect of posterior tenectomy of the superior oblique on the correction of A-pattern, superior oblique overaction and the changes in torsion that occur in such cases. PATIENTS AND METHODS: This prospective study included 15 consecutive cases of bilateral superior oblique overaction with an A pattern of more than 20 PD, a difference of deviation between 25 degrees up-gaze and 35 degrees downgaze, and superior oblique overaction of 2+ to 3+ on a scale of 0 to 4+. Deviation was measured in the primary position, 25 degrees upgaze, and 35 degrees downgaze using the prism bar cover test, and torsion was measured using a synoptophore, the double Maddox rod test, and fundus photographs. Measurements were obtained preoperatively and postoperatively at 1 week, 1 month, and 3 months. All case-patients underwent a standard temporal route posterior tenectomy of the superior oblique by a single surgeon. RESULTS: Mean age was 11.2 +/- 4.2 years with 14 cases of A-pattern exotropia. Mean superior oblique overaction was 2.60 +/- 0.50 in the right eye and 2.26 +/- 0.45 in the left eye, which decreased postoperatively to 2.20 +/- 0.56 and 1.80 +/- 0.41, respectively. The index of surgical effect was 0.84 in the right eye and 0.79 in the left eye. Postoperatively, mean correction of the A-pattern was 17.53 +/- 5.82 PD (index of surgical effect, 0.7). Subjective measurement of torsion was more consistent with the synoptophore compared with the double Maddox rod test. Objective measurement of torsion (fundus photography) was higher compared with subjective measurement. Postoperatively, there was insignificant change in the amount of torsion in upgaze and primary position. CONCLUSION: Posterior tenectomy of the superior oblique results in significant and controlled weakening of the superior oblique and collapse of the A-pattern with a clinically insignificant change in the amount of torsion. PMID- 16250218 TI - Morning glory syndrome: clinical, computerized tomographic, and ultrasonographic findings. AB - OBJECTIVE: To evaluate the ophthalmic, radiologic, and ultrasonographic findings in morning glory syndrome. PATIENTS AND METHODS: Retrospective review of patients' charts. RESULTS: Twenty-one patients were included. Visual acuity was 20/200 or better in 50% of the eyes. Three eyes developed a retinal detachment. When computerized tomography of the orbit demonstrated peripapillary scleral staphyloma and intraocular calcifications, this correlated with poor visual acuity. Computerized tomography of the brain revealed abnormalities in three asymptomatic patients. B-scan ultrasonography demonstrated retinal tissue overhanging the peripapillary scleral staphyloma. CONCLUSIONS: Morning glory syndrome has a spectrum of severity, with most patients retaining useful vision. Orbital and cerebral computerized tomography scan as well as B-scan ultrasonography may help with diagnosis and management. PMID- 16250219 TI - Long-term results of hemihang-back lateral rectus recession. AB - PURPOSE: To report our long-term results with the hemihang-back technique for lateral rectus muscle recession in moderate- and large-angle exotropia. PATIENTS AND METHODS: The medical records of 144 consecutive patients who underwent unilateral or bilateral hemihang-back lateral rectus recession between 1999 and 2002 were reviewed. The amount of surgery performed was based on the distance exotropic deviation. Surgical outcomes were analyzed for all patients included in the study. RESULTS: A total of 66 patients met the inclusion criteria. Sixteen patients had undercorrection, but 6 of these patients redeveloped an exodeviation 6 months after the surgery. No overcorrections were observed in this study. CONCLUSION: The hemihang-back technique is a safe and effective alternative to conventional lateral rectus recession in patients with moderate- or large-angle exotropia. PMID- 16250220 TI - Epiblepharon with inverted eyelashes and high body mass index in Japanese children. AB - PURPOSE: To evaluate the sexual predilection of epiblepharon in Japanese children and the relationship between this condition and body mass index. SUBJECTS AND METHODS: Japanese children who were 6 to 14 years old were cross-sectionally surveyed on routine health screening. RESULTS: Of 2,941 children examined, 127 had epiblepharon. The incidence decreased with age. No sexual predilection was found. Lower eyelids were commonly involved bilaterally. Mild complaints included discomfort or foreign body sensation. The body mass index in children 6 to 11 years old with epiblepharon was higher than that in children without epiblepharon. CONCLUSIONS: No sexual predilection was found for epiblepharon. A high body mass index was noted in children with epiblepharon who were 6 to 11 years old. PMID- 16250221 TI - Repair of a canalicular laceration in a neonate. AB - A neonate with lower eyelid and corresponding canalicular laceration of the left eye was examined 2 hours after birth. The eyelid and canalicular laceration were repaired with a short, 24-gauge catheter as the stent. The laceration healed with minimal scarring, and the epiphora had disappeared by the 12-month follow-up visit. PMID- 16250222 TI - Corneal keloid in Lowe syndrome. AB - We describe a 3-year-old boy with Lowe syndrome who previously underwent bilateral cataract surgery with intraocular lens implantation and strabismus surgery and developed an inferior corneal keloid. The lesion was resected. We report the results of immunohistochemistry analysis of the specimen, as well as alternative treatment modalities for this rare condition. PMID- 16250223 TI - Unilateral Peters' anomaly in a patient with DiGeorge syndrome. AB - We report a case of unilateral Peters' anomaly in a 3-month-old infant with a microdeletion in chromosome 22q11.2, a finding not previously described. This anterior segment anomaly can be explained by a problem in neural crest development, as neural crest cells are known to play a role in the developmental defects of this disorder. PMID- 16250224 TI - Atypical presentations of orbital cysticercosis. AB - We describe three patients with orbital cysticercosis who presented with atypical clinical or radiologic features previously unreported. All three patients had a cyst with a scolex on imaging studies. After 6 weeks of treatment, all three had almost complete resolution of their features. PMID- 16250226 TI - America's best health plans. PMID- 16250225 TI - Sturge-Weber syndrome (choroidal hemangioma and glaucoma). PMID- 16250227 TI - Making them pay. PMID- 16250228 TI - Plans that opt out. PMID- 16250229 TI - The 2005 rankings. How more than 500 health plans were put through the data wringer. PMID- 16250230 TI - [Taxonomical study of bacteria from genera Alteromonas and Pseudoalteromonas isolated from Black Sea water and invertebrates]. AB - Nineteen (19) strains of bacteria have been isolated from the Black Sea water and invertebrates (mollusks and actinia). Most of them have been identified as Alteromonas macleodii, Pseudoalteromonas citrea and P. haloplanktis on the basis of polyphasic taxonomical analysis. Six strains showed 96-97 % similarity to 16S rRNA sequence of the known species of Pseudoalteromonas and obviously belonged to new species. The studied strains have been characterized by a wide spectrum of phenotypic features (morphology, enzyme activity, spectra of carbon nutrition, antibiotic sensitivity); high sensitivity of P. haloplanktis strains to cephalotin and resistance of A. macleodii strains to furadonin made them different than other studied strains of Alteromonas and Pseudoalteromonas. PMID- 16250231 TI - [PCR based identification of strains of endophytic ecotype of Bacillus amyloliquefaciens]. AB - A search for genetic markers of an endophytic ecotype of Bacillus amyloliquefaciens was performed by the subtractive hybridization technique. The isolated fragments specificity was checked by DNA-DNA hybridization. The sequence of a DNA locus exhibiting the highest taxon specificity has manifested a 76% identity to the gene mtlA from Bacillus subtilis 168 genome. A set of discriminative primers has been derived from this sequence which could be used for the fast and reliable identification of isolates of the endophytic ecotype of B. amyloliquefaciens placing them separate against their close relatives including the type cultures of B. amyloliquefaciens and B. subtilis. Identification of strains of this ecotype is quite important as far as their characteristic ability to colonize the roots and internal tissues of plants in association with high antagonistic activity in respect of many phytopathogenic fungi and bacteria makes them valuable agents for plant protection. PMID- 16250232 TI - [Phosphatase activity of Bacillus subtilis IMV B-7023]. AB - Phosphatase activity of two strains of bacteria - Bacillus subtilis IMV B-7023 and B. megaterium 12 is investigated. The phosphatase activity is found to reach 260 mkmol/g x hour for B. subtilis IMV B-7023 and 12-100 mkmol/g x hour for B. megaterium 12 at optimal temperature (55 degrees C) and pH (9.5-10.0). Synthesis of alkaline phosphatase is shown to reach its maximum values at the end of logarithmic phase of the culture growth. It is revealed that Mg2+, Ca2+ cations increase phosphotase activity of B. subtilis IMV B-7023, at the same time Cu2+, Mn2+, Zn2+ cations and inorganic phosphate decrease it. Dependence of the rate of phosphatase reaction of B. subtilis IMV B-7023 on substrate concentration is determined. PMID- 16250233 TI - [Effect of silver nitrate, adenosine-5'-monophosphate and phosphoenolpyruvate on activity of extracellular fructose bisphosphatase of Acholeplasma laidlawii var. granulum strain 118]. AB - The reactions of glycolysis or gluconeogenesis proceed in good coordination in the cells of microorganisms, and each stage of these processes is distinctly regulated. Under such conditions fructose-bisphosphatase (FBPase) activity (the enzyme level being constant in the cells of microorganisms) is inhibited by adenosine-5'-monophosphate (AMP) and is activated by phosphoenolpyruvate (PEP) depending on the kind of the source of carbon (glycolytic or glyconeogenic) used for microorganism growth. It is evident that the corresponding regulation of FBPase should be absent in the extracellular environment where one cannot observe a distinct coordination of functioning of the enzyme systems. The investigation results prove that both AMP and PEP, under their individual testing in concentrations up to 20 microM did not practically affect activity of extracellular FBPase, and at higher concentrations they sharply decreased its activity (200 microM AMP by 70%, and PEP - by 75%). Under joint use of PEP and AMP (in concentration 200 microM and 500 microM) one could observe mutual neutralization of the effect of these substances on FBPase; as a result, its activity decreased only by 15% under AMP concentration of 500 microM, and by 25% at AMP concentration of 200 microM, that is in complete agreement with the data of individual testing of the above substances. PEP in high concentrations has displayed itself as a more active repressor of FBPase activity than AMP. AgNO3 in concentrations to 20 microM has manifested itself as a moderate stimulator of FBPase activity and even in the concentration of 200 microM it decreased the enzyme activity by 50% only. The data obtained are rather different than those described in literature for cellular FBPases of microorganisms. It is known that AMP is a powerful inhibitor of its FBPases activity (Ki = 5 microM) while PEP activates it (Ka = 20 microM). PMID- 16250234 TI - [Effect of plant growth regulators on physiological activity of Bradyrhizobium japonicum ]. AB - Influence of plant growth regulators Ivin, Emistim C, Eney and Agrostimulin on the biomass production and exopolymers synthesis of soybean nodule bacteria, which have contrasting symbiotic properties, and glutamine synthetase activity of their cell-free extracts were studied. It was shown that the processes of the biomass and exopolymers accumulation had an opposite direction. Of all preparations only Ivin and Agrostimulin intensificol growth activity of the microorganisms under study. The level of glutamine synthetase activity and this enzymatic reaction specificity to the bivalent metal ions were determined by the special features of Bradyrhizobium strains and nature of the plant growth regulators. Only in the presence of Eney the increase of glutamine synthetase activity of both cultures of Bradyrhizobium japonicum was established. PMID- 16250235 TI - [Effect of cultivation conditions of Streptomyces globisporus 3-1 on landomycin E production]. AB - The producing of antitumor angucycline antibiotic landomycin E has been studied in dynamics of the surface and deep growth of Streptomyces globisporus 3-1 on different full-value media--soya, maize and peptone-yeast media. It is shown that the highest yield of landomycin E is observed on the maize, a bit lower--on soya, and the lowest--on peptone-yeast media independent of the method of growing. The antibiotic stability is affected by alkali pH of the medium. Growth dynamics of the culture in the maize and soya media in the laboratory 2-litre fermenter is characterized by intensive assimilation of glucose, absorption of dissolved oxygen, accumulation of biomass and landomycin E from the 24th to 48th hour of fermentation. The antibiotic is synthesized as primary metabolite reaching its maximum (180-200 mg/l) for the 48th hour of the culture growth. A hypothesis is put forward on the inhibition of the two-component system of signals transduktion by isoflavone genistein, which stimulates synthesis of landomycin E. PMID- 16250236 TI - [Characteristics of virus double-stranded RNA, isolated from microscopic fungi parasitizing on sugar beet]. AB - We have carried out comparative studies of double-stranded RNA (dsRNA) of viral nature isolated from sugar beet leaves and from mycelium of microscopic fungi using different methods such as PAAG electrophoresis and by polymerase chain reaction (PCR). It was shown that the fragments of dsRNA from sugar beet leaves and from mycelium microscopic fungi had the identical electrophoretic pattern and the same size (1.8 and 2.0 kbp). Using PCR technique it was shown, that isolated dsRNA have a common template for amplification. Electron microscopy of PCR positive mycelium allows us to detect the virus particles of the spherical form with diameter 30-40 nm. The obtained data confirm our previous suppositions, concerning the belonging of isolated dsRNAs (size 1.8 and 2.0 kbp) to new mycovirus targeted a microscopic fungus, instead of beet cryptic viruses. PMID- 16250237 TI - [Detection of antigen-antibody interaction of human adenovirus by the method of surface plasmon resonance]. AB - A possibility to detect adenoviral protein--hexon, using specific antibodies by surface plasmon resonance (SPR) was demonstrated. The hexon of the human adenovirus 2 (Ad2) binds to antibodies immobilized on the sensor surface treated by KNCS and protein A Staphylococcus aureus. The specificity of antihexon antibodies was demonstrated by indirect method of fluorescent antibodies (MFA) and cellular variant of the immunoassay (cELISA). PMID- 16250238 TI - [Kidney bean "Pervomayskaya" as the indicator plant for tobacco mosaic virus]. AB - It was shown that garden beans of "Pervomayskaya" variety respond to mechanical inoculation of leaves with tobacco mosaic virus by formation of local lesions, and thus it may be the indicator plant for the virus. PMID- 16250239 TI - [Adhesive properties of lactic acid bacteria and methods of their investigation]. AB - Current status of the question concerning the study of lactic acid bacteria adhesion process is presented in the review. Lactic acid bacteria colonization capacity allows them to form monolayers, to join the mucosal microbiota, to become a component of ecological barrier, which protects an organism from penetration of pathogens. The available methods of investigation of adhesive properties such as microbiological, light and electron microscopy, biophysical, mathematical and others are presented in detail. The works concerning lactic acid bacteria adhesines nature and factors that influence the exhibition of adhesion have been analyzed. Studies concerning the mechanisms responsible for attachment of cells of lactic acid bacteria to the surface of different types of epithelium and substances that are products of endothelium secretion are analyzed. Questions on adhesion specificity and factors that inhibit its exhibition are considered. The chart of current concept of the adhesive process and factors that reciprocally affect the exhibition of lactic acid bacteria adhesion properties is developed and presented by the authors. PMID- 16250240 TI - [Modern methods for formaldehyde, methanol and ethanol analysis]. AB - It is necessary to create very specific and sensitive methods for assaying formaldehyde and methanol which are produced on the large scale and are very toxic and have mutagenic and carcinogenic action on living organisms. The methods for determination of formaldehyde, methanol and ethanol in the environment and fermentation products published and developed by the authors are reviewed in this paper. Most of the known methods are not sufficiently selective and sensitive and some of them are very expensive. Classical chemical, enzymatic, chemosensor and biosensor approaches used for methanol and formaldehyde assay are described. Enzymatic methods exploiting alcohol oxidase isolated from the mutant over producing strain of methylotrophic yeast Hansenula polymorpha permit efficient determination of formaldehyde in industrial wastewaters. Enzymatic-chemical method based on the use of alcohol oxidase and 4-amino-5-hydrazine-3-mercapto 1,2,4-triazole (AHMT) allows simultaneous determination of methanol and formaldehyde. The technology of biosensor construction and their bioanalytical characteristics are described. Experimental data concerning amperometric and potentiometric biosensors based on the use of genetically modified cells of methylotrophic yeast Hansenula polymorpha are reviewed. The possibility to use alcohol oxidase-based biosensor for the assay of methanol in wastewater is demonstrated. PMID- 16250241 TI - [Plant genetic engineering in Monsanto company: from the first laboratory experiments to worldwide practical use]. AB - The history of modern biotechnology of agricultural plants is briefly considered in the article. Methods of genetic transformation and regeneration of transgenic plants as well as the mechanisms of resistance of genetically modified plants to herbicides and pests are discussed. By the example of genetically modified varieties and hybrids there are shown the ways of solving the problem of weeds and pests. The questions of biosafety legislation in different countries are considered. PMID- 16250242 TI - [Achievements and problems of genetic engineering of Crucifereceae plants]. AB - Plants of the Brassicaceae family are important oil, vegetable and feed crops. The review is devoted to the latest achievements in genetic engineering of plants from this family. Results concerning development of effective methods both of Agrobacteium-mediated transformation and of direct gene uptake are considered. Particularly, possibilities of plant genetic modification with the aim to improve agronomically and commercially important traits are stressed. Problems of biologically safe introduction of transgenic plants into agricultural production are discussed. PMID- 16250243 TI - [Genetic transformation of sugar beet: evolution of theoretical and experimental approaches]. AB - The review is dedicated to several aspects of sugar beet (Beta vulgaris L.) biotechnology: in vitro cultivation, callus induction, plant regeneration and genetic transformation. Media composition, methods of plant regeneration via somatic embryogenesis and protoplast culture are analysed. The use of Agrobacterium tumefaciens and gold particle bombardment is the base for modern genetic transformation methods. PMID- 16250244 TI - Environmental release of living modified organisms: current approaches and case studies. AB - Agricultural biotechnology is being rapidly adopted as evidenced by the acreage of genetically modified (GM) crops planted and tonnes of product (grain and fiber) harvested. Concurrent with this technological progress, is a growing concern that the worlds biological diversity is coming under increasing threat from human activities. As such, ecological risk assessment approaches are being developed for GM crop plants as international agreements regulating the transboundary movements of these products are being implemented. This paper reviews the ecological risk assessment approach that has been used to date to approve GM crops to date. The process has been case-by-case, using a comparative, science-based approach balancing the potential risks and benefits of the new technology versus those present with the currently accepted practices. The approach used to evaluate and approve these products is consistent with the conditions and requirements outlined in the Cartagena Protocol. PMID- 16250245 TI - [Site-specific expression of single-stranded antibody fragments in Nicotiana tabacum]. AB - Antibody expression and immunomodulation are modern molecular techniques to produce pharmaceuticals and to interfere with cellular metabolism or pathogen infectivity in plants. Nonetheless, there is still no generally applicable strategy to express correctly folded active antibodies or antibody fragments in different cell compartments. To facilitate expression, single-chain antibody fragments (scFvs) were made of mouse monoclonal antibodies, J2 and P6 that specifically recognize double-stranded RNA (dsRNA). Stabilizing double-stranded replication intermediates could modulate the biological activity of dsRNAs in plants, especially to influence virus replication. Along with cytoplasmic expression, scFvs were anchored to the plasma membrane; targeted to the apoplast for secretion and made ER-resident. Expression levels were analysed and transgenic plants were evaluated for resistance or tolerance to potato virus Y infection. We have established strategies for expression of correctly assembled antibodies or antibody fragments in different plant cell compartments. PMID- 16250246 TI - [Creation and analysis of Brassica napus + Arabidopsis thaliana somatic hybrids possessing maize Spm/dSpm heterologous transposable system]. AB - Functionally asymmetric somatic hybrids possessing heterologous transposable element Spm/dSpm were obtained following intertribal somatic hybridization between Brassica napus and transgenic Arabidopsis thaliana. Mobile genetic elements actively transposed in the hybrid genomes. Complete elimination of A. thaliana genome was not observed. PMID- 16250247 TI - [Molecular-genetic analysis of wheat (T. aestivum L.) genome with introgression of Ae. cylindrica Host genetic elements]. AB - Wheat-aegilops hybrid plants Triticum aestivum L. (2n = 42) x Aegilops cylindrica Host (2n = 28) were investigated with using microsatellite markers. In two BC1F9 lines some genome modifications connected with losing DNA fragments of initial variety or appearing of Aegilops genome elements were detected. In some investigated hybrids new amplicons lacking in parental plants were found. Substitution of wheat chromosomes for aegilops chromosomes was not revealed. Analysis of microsatellite loci in BC2F5 plants showed stable introgression of aegilops genetic elements into wheat; elimination of some transferred aegilops DNA fragments in the course of backcrossing; decreasing size of introgressive elements after backcrossing. Introgressive lines were classified according to genome changes. PMID- 16250248 TI - Genetic analysis of the T. aestivum/Ae. sharonensis introgressive lines of common wheat for resistance to powdery mildew. AB - Results of investigation of peculiarities of common wheat hybridological analysis for a discrete character, resistance for powdery mildew, governed by the alien gene from Ae. sharonensis are present. Relation between genome structure of crossed introgressive lines and deviation of empirical ratios of segregation in F2 from theoretical, based on the assumption about monogenic inheritance of considered character is established. The approach to the quantitative count of influence of such connection on distortion of actual segregation in comparison with theoretically expected ratio is developed. PMID- 16250249 TI - [Antimutagenic activity of Armoracia rusticana, Zea mays and Ficus carica plant extracts and their mixture]. AB - Antimutagenic action of plant extracts of Armoracia rusticana, Ficus carica, Zea mays and their mixture on environmental xenobiotics has been investigated. The plant extracts and their mixture decreased the level of mutations induced by N metil-N'-nitro-N-nitrozoguanidin (MNNG) in Vicia faba cells, chlorophyll mutations in Arabidopsis thaliana and NaF induced mutability in rat marrow cells. The studied plant extracts and their mixture demonstrate the ability to decrease the genotoxicity of environmental mutagens. PMID- 16250250 TI - Disposition of paclitaxel (Taxol) and its metabolites in patients with advanced breast cancer (ABC) when combined with trastuzumab (Hercpetin). AB - Monoclonal antibodies are capable of modulating drug metabolising enzymes resulting in unexpected plasma concentrations of a drug when given concomitantly. Therefore plasma concentration of paclitaxel (PTX) and its metabolites has been monitored in 10 patients with advanced breast cancer during treatment with PTX alone or combined with trastuzumab (TMAB, paired cross over design). Compared to the MONO regimen PTX peak plasma concentrations were about 25% lower in the TMAB schedule: cmax = 3294 +/- 1174 ng/ml (MONO: 4368 +/- 1887 ng/ml). TMAB also caused lower peak plasma concentration of the main metabolite 6-hydroxy PTX (248 +/- 89 ng/ml) compared to the MONO schedule (194 +/- 82 ng/ml). Cmax of the minor metabolites was distinctly below 100 ng/ml and consequently differed negligible in both schedules. The similar apparent formation rate of the metabolites in both schedules (range from 30 to 50 min) as well as identical tmax values (range 170 190 min) suggested that TMAB had no influence on PTX metabolism. In accordance to plasma concentrations, AUClast of PTX was lower in the MONO schedule (733 +/- 197 microg/ml*min, AUClast = 669 +/- 248 microg/ml*min for TMAB) but without significance. In summary no indices for an altered plasma disposition of PTX and its metabolites could be found when TMAB was given concomitantly. PMID- 16250251 TI - Metabolism and excretion of the antiepileptic/antimigraine drug, Topiramate in animals and humans. AB - The metabolism and excretion of 2,3:4,5-bis-O-(1-methylethylidene)-beta-D fructopyranose sulfamate (TOPAMAX, topiramate, TPM) have been investigated in animals and humans. Radiolabeled [14C] TPM was orally administered to mice, rats, rabbits, dogs and humans. Plasma, urine and fecal samples were collected and analyzed. TPM and a total of 12 metabolites were isolated and identified in these samples. Metabolites were formed by hydroxylation at the 7- or 8-methyl of an isopropylidene of TPM followed by rearrangement, hydroxylation at the 10-methyl of the other isopropylidene, hydrolysis at the 2,3-O-isopropylidene, hydrolysis at the 4,5-O-isopropylidene, cleavage at the sulfamate group, glucuronide conjugation and sulfate conjugation. A large percentage of unchanged TPM was recovered in animal and human urine. The most dominant metabolite of TPM in mice, male rats, rabbits and dogs appeared to be formed by the hydrolysis of the 2,3-O isopropylidene group. PMID- 16250252 TI - Pharmacokinetics of scutellarin and its aglycone conjugated metabolites in rats. AB - Scutellarin, a flavonoid glycoside, is the primary active ingredient in breviscapine that is a mixture of flavonoid glycosides extracted from a Chinese herb Erigeron breviscapus (Vant.) Hand.-Mazz. The pharmacokinetics of scutellarin and its aglycone conjugated metabolites after intraperitoneal injection and oral administration of breviscapine was investigated in rats. The plasma concentration of scutellarin and scutellarein conjugates in serial samples was measured by validated high-performance liquid chromatography methods. The pharmacokinetic results indicated that scutellarin underwent rapid and extensive biotransformation in vivo. After intraperitoneal injection, scutellarin was absorbed rapidly. The profiles of scutellarin and scutellarein conjugates were fitted to a two-compartment open model. Scutellarin and scutellarein conjugates showed a similar time course. No significant difference in tmax, t0.5(alpha) and t0.5(beta) was observed between scutellarin and scutellarein conjugates (p>0.05). After oral administration, fluctuations were observed in the concentration-time profiles of both scutellarin and scutellarein conjugates and the pharmacokinetics could not be explained by classical compartment model. The relative bioavailability of oral administration was very low, 10.67 % +/- 4.78% for scutellarin and 7.92% +/- 1.90% for scutellarein conjugates. PMID- 16250253 TI - Alterations in cefalosporin levels in the serum and mandible of hyperlipaedemic rats after co-administration of ibuprofen. AB - Several interactions between antibiotics and non-steroidal anti-inflammatory drugs (NSAIDs) have been described in the literature, and it has been reported that hyperlipidaemia induces significant changes in cefalosporin levels. The aim of this study was to determine the changes in the levels of several cefalosporins in the serum and mandible after ibuprofen co-administration in hyperlipaedemic rats. One hundred and forty male Wistar rats were used and divided in 4 groups (A D), each of which was further divided into 5 subgroups (1-5), either with placebo or with various treatment regimes. The co-administration of NSAIDs led to increased cefalosporin levels in both control and hyperlipidaemic animals. Hyperlipidaemia was also found to augment cefalosporin levels. These observed increases might be due to the displacement of the cephalosporins from their binding sites in serum albumin and tissue proteins in the presence of ibuprofen. NSAIDs showed a greater binding affinity for tissue proteins compared to the cephalosporins, and probably play an antagonistic role in protein binding, resulting in higher concentrations of antibiotics. PMID- 16250254 TI - Pharmacokinetics of TJ-8117(Onpi-to), a drug for renal failure (II): effects of food, repeated administration and renal failure to plasma concentration of [3H]-( )epicatechin 3-O-gallate in rats. AB - TJ-8117 (Onpi-to) is a herbal medicine extracted from mixture of five crude drugs (Rhei Rhizoma, Glycyrrhizae Radix, Ginseng Radix, Zingiberis Rhizoma and Aconiti Tuber), which has been developed as a drug for chronic renal failure. ( )Epicatechin 3-O-gallate (ECG), one of the active compounds of TJ-8117, was labeled with tritium and spiked to TJ-8117. Effects of food, renal failure and repeated administration to pharmacokinetics of ECG-related radioactivity in the plasma were investigated after oral administration of TJ-8117 containing [3H]ECG ([3H]TJ-8117) in male rats. After oral administration of [3H]TJ-8117, the radioactivity in the plasma in non-fasted rats was higher than that in fasted rats. AUC(0-72 h) and Cmax in the non-fasted was 123% and 248% of those in the fasted. After oral administration of [3H]TJ-8117, the radioactivity in the plasma in 5/6 nephrectomized rats was higher than that in sham-operated rats. AUC(0-72 h) and Cmax in 5/6 nephrectomized was 332% and 236% of those in sham-operated. After repeated administration of TJ-8117 for 6 days, [3H]TJ-8117 was administered on 7th day. Radioactivity in plasma in first day was similar to that in 7th day. The pharmacokinetic parameters were not significantly different between single and repeated administration. PMID- 16250255 TI - A comparison of glutathione S-transferase mutant frequencies in healthy Han and Uygur Chinese. AB - OBJECTIVE: This study was to compare the frequencies of genetic polymorphisms of GSTM1, GSTT1, and GSTP1 in Uygur Chinese with those in Han Chinese. METHODS: GSTM1 and GSTT1 polymorphisms were analyzed by a PCR-Multiplex procedure, whereas GSTP1 polymorphism was analyzed by PCR-RFLP. RESULTS: The frequency of GSTM1 null genotype in Han Chinese (56.1%) was similar to that in Uygur Chinese (53.2%) (P = 0.592), whilst the frequency of GSTT1 null genotype in Han Chinese (50.0%) was significantly (P < 0.05) higher than that of Uygur Chinese (26.6%). GSTP1 had a genotype distribution of 60.7% I/I, 35.2% I/V and 4.1% V/V in Han Chinese, and 51.3% I/I, 40.2% I/V and 8.4% V/V in Uygur Chinese. CONCLUSION: There is marked ethnic difference in the mutant frequencies of GSTT1 and GSTP1, but not GSTM1, between Uygur and Han Chinese. PMID- 16250256 TI - Assessment of oral bioavailability and preclinical pharmacokinetics of DRF-6196, a novel oxazolidinone analogue, in comparison to linezolid. AB - The aim of this study was to determine the bioavailability of a novel oxazolidinone, DRF-6196, in mice and rats following intravenous (i.v) and oral dosing and to compare the pharmacokinetics with those obtained following linezolid dosing. Blood samples were drawn at predetermined intervals up to 24 h post-dose after either DRF-6196 or linezolid administration. The concentrations of DRF-6196 and linezolid in various plasma samples were determined by a HPLC method. Following oral administration maximum concentrations of DRF-6196 were achieved within 0.5 h irrespective of the species. While the doses increased in the ratio of 1 : 3 : 10, mean Cmax and AUC(0-infinity) values in mice for DRF 6196 increased in the ratio of 1 : 3.87 : 8.53 and 1 : 2.51 : 9.24, respectively. Both the Cmax and AUC(0-infinity) values increased almost proportional to the dose administered in mice. Following i.v administration, the concentration of DRF 6196 declined in a bi-exponential fashion with terminal elimination half-life of 1.5 h irrespective of the species. The systemic clearance and volume of distribution of DRF-6196 in mice were 1.14 L/h/kg and 0.66 L/kg, respectively after i.v administration, while the respective values in rats were 0.61 L/h/kg and 0.41L/kg, respectively. Elimination half-life ranged between 0.8-1.5 h. Absolute oral bioavailability of DRF-6196 was found to be 80-96% across the test dose range. Although plasma levels of DRF-6196 were lesser compared to linezolid in the initial hours, it may not have any consequences on the clinical effectiveness of the molecule. PMID- 16250260 TI - Deciphering the sinus tachycardias. PMID- 16250258 TI - Single and chronic administration of ciprofibrate or of ciprofibrate-glycinate in male Fischer 344 rats: comparison of the effects on morphological and biochemical parameters in liver and blood. AB - Fibrates lead to a reduction of serum triglycerides and cholesterol in hyperlipidemic patients. Their therapeutic use, however, can be associated with adverse effects like gastrointestinal disorders, myalgia, myositis and hepatotoxicity. Large doses can even cause hepatocellular carcinoma in rodents. Additionally, interactions with the biotransformation of other compounds at the cytochrome P450 (CYP) system have been observed. Thus, the discovery of new derivatives with less of these side effects is of great interest. In the present study a single (10 mg/kg body weight) or a 4-week (1 or 10 mg/kg body weight daily) oral administration of ciprofibrate or of the newly synthesized ciprofibrate-glycinate was investigated in adult male Fischer 344 rats. Serum lipid concentrations were distinctly decreased after single but only slightly after chronic administration of the two fibrates, whereas liver parameters revealed a slight concentration and time dependent hepatotoxicity. Histologically, a hypertrophy, an eosinophilia, a reduced glycogen content and also an apoptosis of the hepatocytes was observed. Effects were more pronounced after chronic treatment and after application of the higher dosage. All CYP enzymes investigated were induced in a time and concentration dependent manner. Resulting CYP mediated monooxygenase and oxidase activities showed a dependency both on enzyme induction and hepatotoxic effects. With no parameter investigated major differences were seen between ciprofibrate and ciprofibrate-glycinate. Thus, the present investigations revealed no noticeable advantages of ciprofibrate-glycinate over its parent compound ciprofibrate. PMID- 16250257 TI - Effect of methamphetamine on the pharmacokinetics of dextromethorphan and midazolam in rats. AB - Methamphetamine is the fourth most frequently reported compound associated with drug abuse on admission of patients to treatment centres after cocaine, heroin and marijuana. It is metabolized in the organism with a reaction that is catalyzed by cytochrome P450, mainly by the CYP2D and CYP3A subfamily, 4 hydroxyamphetamine and amphetamine being dominant metabolites. The present pharmacokinetic study was undertaken to investigate the possible influence of methamphetamine (10 mg/kg, i.p., once daily for six days) on the pharmacokinetics of dextromethorphane as a model substrate for rat cytochrome P-4502D2 and midazolam as a model substrate for CYP3A1/2. Animals received a single injection of dextromethorphane (10 mg/kg) or midazolam (5 mg/kg) in the tail vein 24 h after the last dose of methamphetamine or administration of placebo. The results of pharmacokinetic analysis showed a significantly increased rate of dextrorphane and 3-hydroxymorphinan formation, and a marked stimulatory effect of methamphetamine on CYP2D2 metabolic activity. Similarly, the kinetics of midazolam's metabolic conversion to hydroxy derivates of midazolam indicated a significant increase in CYP3A1/2 activity. The results showed that the administration of methamphetamine significantly stimulated the metabolic activity of CYP2D2 as well as that of CYP3A1/2. With regard to the high level of homology between human and rat CYP isoforms studied, the results may have a clinical impact on future pharmacotherapy for methamphetamine abuse. PMID- 16250261 TI - Teaching proper cardiovascular physical diagnosis. PMID- 16250259 TI - Influence of famotidine on verapamil pharmacokinetics in rats. AB - The effect of famotidine (4 mg/kg, p.o.) on the kinetic profile of co administered verapamil (20 mg/kg(-1), p.o.) was studied in the rat. Plasma verapamil levels were collected serially for 12 h and measured using sensitive HPLC method. The pharmacokinetic parameters (elimination half-life, area under the plasma concentration-time curve, peak plasma levels and the times to attain these plasma levels) of verapamil were evaluated in the rat. The results indicate that co-administered famotidine did not significantly alter the pharmacokinetic profile of verapamil in the rat. The present finding suggests that famotidine may safely be co-administered with verapamil but clearly further studies in human subjects are needed to reliably rule out the potential interaction of these two drugs. PMID- 16250262 TI - Is aggressive lipid-lowering effective and safe in the older adult? AB - With improvements in diet and health care over the last several decades, the elderly have become the fastest-growing segment of the U.S. population. Along with this progress, however, has come increasing morbidity, much of which is attributable to cardiovascular disease, and which is burdensome both to patients and to the health-care system. New medications, including HMG-CoA reductase inhibitors (statins) have the potential to prevent or delay the progression of disease and vascular events. Large, randomized, placebo-controlled trials continue to show that elderly patients benefit from statin therapy and that aggressive therapy is generally very safe in this population; however, statins remain under-used in older individuals. This paper reviews the benefits of statin therapy in the elderly and discusses the reality versus the perception of risk, selection of appropriate patients, and ways in which any risks that may exist can be minimized further. PMID- 16250263 TI - High-sensitivity C-reactive protein as a risk assessment tool for cardiovascular disease. AB - Almost half of first cardiovascular events occur in individuals with no known risk factors. Attempts in the last decade to predict cardiovascular risk more accurately have led to the emergence of a novel risk factor, C-reactive protein (CRP), which has proved to be as good a risk predictor as low-density lipoprotein cholesterol. C-reactive protein is an index of inflammation that is now believed to promote directly all stages of atherosclerosis, including plaque rupture. As measured by high-sensitivity assays, high-sensitivity CRP (hs-CRP) also independently predicts recurrent events in patients with known coronary artery diseases. Recent evidence implicates hs-CRP, and thus inflammation, in the metabolic syndrome and diabetes mellitus, particularly in women. As a clinical tool for cardiovascular risk assessment, hs-CRP testing enhances information provided by lipid screening or global risk assessment. Statin therapy and other interventions can lower hs-CRP. Whether or not such reductions can prevent cardiovascular events is under investigation. PMID- 16250264 TI - Cholesterol crystals cause mechanical damage to biological membranes: a proposed mechanism of plaque rupture and erosion leading to arterial thrombosis. AB - BACKGROUND: Plaque rupture and/or erosion is the leading cause of cardiovascular events; however, the process is not well understood. Although certain morphologic characteristics have been associated with ruptured plaques, these observations are of static histological images and not of the dynamics of plaque rupture. To elucidate the process of plaque rupture, we investigated the transformation of cholesterol from liquid to solid crystal to determine whether growing crystals are capable of injuring the plaque cap. HYPOTHESIS: We hypothesized that during cholesterol crystallization the spatial configuration rapidly changes, causing forceful expansion of sharp-edged crystals that can damage the plaque cap. METHODS: Two experiments were performed in vitro: first, cholesterol powder was melted in graduated cylinders and allowed to crystallize at room temperature. Volume changes from liquid to solid state were measured and timed. Second, thin biological membranes (20-40 microm) were put in the path of growing crystals to determine damage during crystallization. RESULTS: As cholesterol crystallized, the peak volume increased rapidly by up to 45% over 3 min and sharp-tipped crystals cut through and tore membranes. The amount of cholesterol and peak level of crystal growth correlated directly (r = 0.98; p < 0.01), as did the amount of cholesterol and rate of crystal growth (r = 0.99; p < 0.01). CONCLUSIONS: These observations suggest that crystallization of supersaturated cholesterol in atherosclerotic plaques can induce cap rupture and/or erosion. This novel insight may help in the development of therapeutic strategies that can alter cholesterol crystallization and prevent acute cardiovascular events. PMID- 16250265 TI - Effects of fluvastatin, an HMG-CoA reductase inhibitor, on serum levels of interleukin-18 and matrix metalloproteinase-9 in patients with hypercholesterolemia. AB - BACKGROUND: Interleukin-18 (IL-18), a novel proinflammatory marker, and matrix metalloproteinase-9 (MMP-9) represent the indices of plaque stability. It is unknown whether hydroxymethylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins), which provide anti-inflammatory and endothelium protection effects, have the property of stabilizing plaque in patients with hypercholesterolemia. HYPOTHESIS: The study was designed to investigate the influence of statin therapy in circulating IL-18, MMP-9, and endothelial function. METHODS: We investigated the effects of a 12-week therapy with fluvastatin on IL-18, MMP-9, and endothelial function in patients with hypercholesterolemia. RESULTS: Compared with placebo, fluvastatin significantly improved flow-mediated vasodilatation to hyperemia, a hallmark of endothelial function [from 3.8% (-3.9 approximately 15.2) to 5.9% (-0.3 approximately 13.2), p = 0.001], and attenuated plasma levels of high sensitivity C-reactive protein (hsCRP) [from 1.3 (0.3 approximately 7.7) to 1.1 mg/l (0.2 approximately 3.5), p = 0.018], IL-18 [from 247.6 (145.4 approximately 378.4) to 196.4 pg/dl (90.7 approximately 380.2), p <0.001], total MMP-9 (from 58 +/- 46.3 to 39.4 +/- 22.4 ng/dl, p = 0.023), and MMP-9 activity [from 6.4 (3.6 approximately 27) to 5.6 ng/dl (3.1 approximately 13.7)]. However, no significant correlation was found between the degree of changes in lipid profile and flow-mediated dilatation (FMD) and plasma concentration of IL-18 and MMP-9. CONCLUSIONS: Fluvastatin reduced plasma concentrations of IL-18 and MMP-9, and improved endothelial function in patients with hypercholesterolemia independent of its lipid-lowering effect. PMID- 16250266 TI - Procedural success and complications following percutaneous coronary interventions among Asians and Pacific Islanders. AB - BACKGROUND: Although Asians and Pacific Islanders (PI) make up the fastest growing ethnic group in the United States, little is known about the clinical characteristics, procedural success, and procedure-related complications of those who undergo percutaneous coronary interventions (PCI). HYPOTHESIS: This study investigated PCI procedural success and procedural complications among PI and Asian patients in comparison with Caucasians. METHODS: We examined clinical characteristics, procedural success (post-PCI lesion < 50%) and procedure-related complications (hemorrhage, renal failure, myocardial infarction, stroke, bypass surgery, death) for all patients undergoing PCI at our hospital from January 1999 to June 2003. RESULTS: Overall, 2,598 PCIs were performed--1,058 (39%) in Caucasians, 1,163 (43%) in Asians, and 377 (14%) in PIs. The mean age of PIs (59 +/- 11 years) was significantly lower than that of Caucasians (65 +/- 12 years) and Asians (66 +/- 12 years). The mean body mass index (26 +/- 5) of Asians was significantly lower, while that of PIs (31 +/- 7) was significantly higher than that of Caucasians (28 +/- 6). More Asians (33.3%) and PIs (40.5%) had diabetes mellitus than did Caucasians (19.9%). More Asians (71.6%) and PIs (76.1%) had hypertension than did Caucasians (61.9%). Renal failure was more prevalent in Asians and PIs (6.0 and 7.4%, respectively) than in Caucasians (3.8%). Other than a higher prevalence of disease involving the left anterior descending vessel in Asians (56.4%) compared with Caucasians (50.4%), angiographic features across the three races were similar. There was no significant difference in procedural success (approximately 94%) or procedure-related complications among Caucasians (6.4%), Asians (7.1%), and PIs (4.3%). CONCLUSION: Although PIs and Asians have a substantially higher burden of comorbidities than Caucasians, race does not appear to influence PCI procedural success or procedure-related complications. PMID- 16250267 TI - Peripheral vascular endothelial function correlates with exercise capacity in women. AB - BACKGROUND: Vascular endothelial function has been observed to correlate with exercise capacity in predominantly male populations. Gender-based differences exist in the clinical course of coronary artery disease, and previous studies indicate that estrogen may influence endothelial function. These observations raise the possibility that the relationship between endothelial function and exercise capacity in women may differ from that in men. HYPOTHESIS: This study aimed to determine whether peripheral vascular endothelial function correlates with exercise capacity in women. METHODS: Women who were referred for clinically indicated exercise testing with technetium-99 myocardial perfusion imaging were consecutively recruited. To ensure a population free of exercise limitation due to ischemic heart disease, women without myocardial perfusion defects were included for analysis in this study (n = 105). Endothelial function was assessed by brachial artery ultrasound flow-mediated vasodilation (FMD). Exercise capacity was defined as the duration of exercise on a symptom-limited Bruce protocol. RESULTS: Mean FMD was 11.8 +/- 0.6%, and median FMD was 12%. Subjects with an FMD less than the median of 12% had a significantly shorter exercise time than those with FMD > or = 12% (411 +/- 17 vs. 482 +/- 24 s, p = 0.014). There was a significant correlation between FMD and exercise time (r = 0.34, p < 0.001). Age and body mass index were additional predictors of exercise time; however, the relationship between FMD and exercise time was independent of these variables. CONCLUSION: Brachial artery FMD correlates with exercise capacity in women, even in the absence of ischemic heart disease. PMID- 16250268 TI - Clinical predictors and characteristics of patients with chronic liver disease and intrapulmonary shunts. AB - BACKGROUND: Intrapulmonary shunting (IPS) is a well-described phenomenon in chronic liver disease but its significance is not known. HYPOTHESIS: The study was undertaken to enhance our understanding of the characteristics and prevalence of IPS. METHODS: We retrospectively studied 204 consecutive patients with end stage chronic liver disease who underwent a dobutamine stress echocardiogram, along with a saline contrast bubble study, as part of their pretransplant evaluation. RESULTS: Intrapulmonary shunting of any degree was present in 56.4% of patients. Patients with IPS were more likely to report alcohol use and less likely to have diabetes mellitus. Patients in Child-Pugh classification C were more likely to have IPS than those with classification A or B. The resting room air PaO2 levels were significantly lower in patients with grade 3-4 IPS than in those with grade 1-2. Elevated estimated pulmonary systolic pressure on echocardiography was more prevalent in patients with than in those without IPS. CONCLUSIONS: A majority of patients with end-stage chronic liver disease undergoing transplant evaluation have IPS. There are important baseline differences between patients with and without IPS. The presence of IPS is associated with increased severity of chronic liver disease. Further study is required to define the prognostic significance of IPS and its impact on future liver transplantation. PMID- 16250269 TI - "Lead tube"-like aorta with intractable systolic hypertension. PMID- 16250271 TI - Frog-marching America to a saner health system. PMID- 16250270 TI - Friedrich Hermann Stannius. PMID- 16250272 TI - Taking stock of two decades of the HIV/AIDS epidemic in Malaysia. PMID- 16250273 TI - A review of day care ureteroscopy of a teaching hospital in Malaysia. AB - This is a retrospective review of 200 day-care ureteroscopies performed between April 1998 and October 2002. The aim was to determine the feasibility and safety of this procedure. The mean age was 38.8 years and 75% of them were male. Eighty three percent were ASA 1 and the remainders were ASA 2. The side of procedure were 48.5% right, 2.5% left and 6% both sides. Fentanyl and proprofol were used during induction. Calculi were found in 82.5% of cases. Rigid and flexible scope were used in 91.5% and 10.5% of cases. There were 33% upper, 23% mid and 44% distal ureteric stones. The mean stone size was 9.37 X 6.93mm. Seventy-four percent required a ureteric stent insertion. The mean operative time was 58.3 minutes. Four percent of patients experienced pain post-operatively relieved by NSAID injection. Repeat ureteroscopy performed in 30% of patients. There were zero admission rates. This study suggests day-care ureteroscopy is effective, safe and well tolerated by patients. PMID- 16250274 TI - Awareness of post-exposure prophylaxis guidelines against occupational exposure to HIV in Hospital Sungai Petani. AB - Occupational risk of Human Immunodeficiency Virus (HIV) infection is a matter of concern for health care workers. We conducted a study to gauge the level of awareness amongst HCW (doctors and nurses) working in Hospital Sungai Petani regarding the post-exposure prophylaxis in case of needle stick injuries from confirmed or suspected cases of HIV. Nineteen Doctors (56%) and 13 nurses (25%) were aware of correct risk of transmission. None identified all the four risk fluids correctly. 94% of doctors and 98% of nurses correctly stated that washing the site with soap and water is the initial procedure, but only few (1/10 of doctors and 1/3 of nurses) knew whom to contact immediately after injury. Twenty three doctors (67%) and 41(78%) nurses were aware of the use of Zidovudine but only 10 participants were aware of the use of second drug. Only 6 doctors (17%) and 8 nurses (15%) knew the correct duration of post-exposure prophylaxis. Twenty three doctors (67%) and 35 nurses (67%) knew that the drugs were available in Hospital Pharmacy and 11 doctors and 12 nurses knew the approximate cost of therapy. On the average about 50% of doctors and nurses have fair knowledge of post exposure prophylaxis against HIV. Ongoing awareness and training are necessary to improve the same. PMID- 16250275 TI - Audit on radiographs in anterior shoulder dislocations. AB - The radiographs of 115 anterior shoulder dislocations (100 patients; 74 males, 26 female were reviewed to assess the radiographic views used in the management of this dislocation. Eighty-eight patients (88%) had only the anteroposterior (AP) view, 10 patients had 2 radiographic views taken and only 2 patients had three radiographic views. Hill-Sachs lesions were found in 18%, and glenoid rim fractures in 3% of the patients. A greater tuberosity fracture was found in 18% of the patients. Therefore, the current practice in the management of an acute anterior shoulder dislocation appears to be to perform a single view (AP) pre reduction radiograph to confirm the diagnosis and a single view (AP) post reduction radiograph to confirm reduction after a close manipulative reduction has been performed. This practice is likely to result in an underestimate of associated Hill Sachs lesion and glenoid rim fractures, but not greater tuberosity fractures. PMID- 16250276 TI - Sensitivity and specificity of portable transient otoacoustic emission (TEOAE) in newborn hearing screening. AB - Early identification and management of hearing impairment is very valuable. The goal standard measurement of hearing loss is by brainstem evoked response (BSER). This prospective study was conducted in Hospital University Kebangsaan Malaysia (HUKM) to determine the sensitivity and specificity of transient evoked otoacoustic emission (TEOAE) as a screening tool for hearing impairment from February 1999 to February 2000. One hundred and thirty-three newborns from postnatal ward and seventy-eight newborns from neonatal intensive care unit (NICU) were screened for possible hearing loss using portable TEOAE. This study showed that TEOAE is a very sensitive but moderately specific screening tool. PMID- 16250277 TI - The feasibility, reliability and validity of the Malay McGill Quality of Life Questionnaire--Cardiff Short Form (MMQOL-CSF) in Malaysian advanced cancer population. AB - Health-related quality of life (HRQoL) assessment is important in healthcare outcomes. This study aimed to determine the feasibility, reliability and validity of the Malay McGill Quality of Life Questionnaire--Cardiff Short Form (MMQOL-CSF) in advanced cancer population. Patients either completed the MMQOL-CSF alone or in addition to its long version. The study recruited 116 participants (average age = 44 years old). On average, MMQOL-CSF was completed in 5.4 minutes. Most domains showed evidence of reliability (Cronbach's alpha = 0.76-0.92). Correlation with its long version was moderate to strong (r(s) = 0.54-0.87). The MMQOL-CSF was a feasible, reliable and valid HRQoL instrument in this population. PMID- 16250279 TI - Attitudes of medical students towards euthanasia in a multicultural setting. AB - A cross-sectional survey of 400 medical students of multicultural backgrounds at the University of Malaya was conducted to understand their attitudes towards euthanasia and factors related to medical decisions and ethical reasoning concerning the prolongation of life, the right to die and euthanasia. The student respondents completed self-administered questionnaires that comprised of twelve questions with multiple stems addressing personal perceptions, knowledge, attitudes, and decisions about euthanasia and the relief of suffering. The majority of respondents (52%) were for the withdrawal of active therapy in a patient suffering from a terminal painful disease while 48% of them were against it. Seventy-one percent of the students involved in the study were against the idea of active euthanasia i.e. the administration of a lethal injection. However, 27% of the respondents felt that there was a moral justification to assist patients to die. Thirty-two percent of the respondents favoured the legalization of euthanasia in Malaysia while 67% of them were strongly against it. The majority (61%) of respondents would not practice euthanasia as a doctor nor would they have performed on themselves if or when it became legal. The main issue surrounding euthanasia that concerned the respondents was the misuse of it by unethical practitioners and they felt that further debate on the matter was essential, both within the local and international communities. PMID- 16250278 TI - Apoptosis in endometria of dysfuntional uterine bleeding women. AB - The study of apoptosis in endometrium of women with irregular uterine bleeding and its predictive value in endometrial malignancy. Analyze apoptotic and mitotic indices and their relevance in irregular uterine bleeding. To determine the expression of Bcl-2 oncoprotein in endometrial glands from patients with irregular uterine bleeding. Department of pathology in a Government Hospital serving a varied socio-economic population in Chennai. Random samples of endometrial currettings from dysfunctional uterine bleeding (DUB) patient who underwent endometrial curettage as therapeutic and diagnostic procedure during the year 2000. Of 50 cases of endometrial samples from patients diagnosed as cases of DUB, the apoptotic and mitotic indexing was carried out and histological categorization revealed 13 cases as Anovulatory. 14 as simple hyperplasia, 5 as early secretory endometrium, 4 as mid secretory and 4 as late secretory endometrium and 7 as endometrium showing features of hormonal imbalance. Three cases were not included, due to sub-optimal processing. A good correlation of the Bcl-2 expression and the apoptotic cell morphology/indices, in the different categories of the endometria of DUB cases is observed. This preliminary study gives an insight to the existence of a correlative pattern of apoptosis in DUB cases. A prospective study on a larger number of cases may substantiate the hypothesis that the Apoptotic and Mitotic indices are useful screening methods with predictive values on development of endometrial carcinoma. It is observed that an increased apoptotic index correlating with high Bcl-2 expression, reflecting the actual cell burden. This prolonged cell survival resisting cell deletion is associated with irregular uterine bleeding endometria. PMID- 16250281 TI - Cystic fibrosis in Malaysian children. AB - Cystic fibrosis (CF) is an autosomal recessive disease commonly found among the Caucasian population. The availability of sweat test and with increasing experience have made it possible to diagnose more cases of CF. Our first case of CF was diagnosed 16 years ago and to date we have managed sixteen cases of CF. Sixteen children were diagnosed with CF in our units at the Paediatric Institute and University Malaya Medical Centre (UMMC). They were referred with either one or all of the following symptoms: i) recurrent pneumonia, ii) bronchiectasis, iii) failure to thrive, iii) malabsorption or iv) history of meconium ileus obstruction during the neonatal period. When the clinical features suggested strongly of CF, sweat tests will be performed in duplicates and considered positive when the sweat chloride or sweat sodium was more than 60 mmol/l for both results. Seventy- two hours fecal fat excretion or stool for fat globule was performed to document malabsorption. From the year 1987 to 2003, 16 patients were confirmed to have cystic fibrosis in Malaysia by positive sweat tests. Thirteen patients were diagnosed in Paediatric Institute while the remaining three were diagnosed in UMMC. On follow-up two patients died due to severe bronchopneumonia at the age of two years old. Although once considered rare, CF should now be considered in any children with clinical presentations of recurrent chest infections, bronchiectasis, in the presence or absence of malabsoption stmptoms and in neonates with meconium ileus obstruction. PMID- 16250280 TI - Clinical predictors of a positive troponin T test in patients presenting with probable acute coronary syndromes. AB - Acute coronary syndrome (ACS) patients with positive troponin T (TnT) test are at higher risk for death and myocardial reinfarction. They would significantly benefit from early aggressive pharmacologic and invasive therapy. However, TnT test is not widely available. This retrospective study of 173 patients with ACS showed: that prolonged or repetitive episodes of angina at rest in the previous 24 hours (p = 0.01) and evidence of myocardial ischaemia on ECG (p < 0.001) were associated with positive TnT tests (> or = 0.1 ng/mL). The two variables in combination showed 100% positive predictive value, facilitating early identification and streamlining of therapy. PMID- 16250282 TI - Pulmonary disease empirically treated as tuberculosis-a retrospective study of 107 cases. AB - Pulmonary disease is sometimes treated empirically as tuberculosis (TB) in the absence of microbial confirmation if the clinical suspicion of active TB is high. In a country of relatively high TB and low HIV burden, we retrospectively studied 107 patients (69.2% male; mean age (SD): 45 (17) years) who received empirical anti-TB treatment for intrapulmonary opacities or pleural effusions suspected of active TB in our hospitals between 1998 and 2002. The diagnosis of definite or probable 'smear-negative' pulmonary TB was made based on treatment outcome at two months with rifampicin, isoniazid, pyrazinamide and ethambutol (or streptomycin). At this end-point, 81 patients (84.4%) had both clinical and radiological improvement (definite cases), 12 (12.5%) had clinical improvement alone and 3 (3.1%) had radiological improvement alone (probable cases). Confirmation of acid fast bacilli was subsequently obtained in 12 patients (all definite cases) from culture of initial pulmonary specimens. Eleven patients (10.5%) were diagnosed as 'non-TB' based on absence of both clinical and radiological improvement or discovery of another cause for the pulmonary condition at or before this two month study end-point. In the 'non-TB' group, 2 had carcinoma, 2 had HIV-related pulmonary diseases, 1 had bronchiectasis, while in 6 causes were indeterminate. Six (6.3%) and 3 (27.3%) patients reported adverse effects from anti-TB drugs from the 'TB' and 'non-TB' groups respectively. Our findings suggest that empirical anti-TB treatment is an acceptable practice if clinical suspicion is high in patients coming in our region. PMID- 16250283 TI - Tarsometatarsal joint injuries--review of clinical presentation and surgical treatment. AB - The tarsometatarsal (Lisfranc) joint suffers a spectrum of injuries ranging from solely ligamentous injury to fracture dislocation. The authors evaluated eighteen consecutive patients who underwent surgery for tarsometatarsal joint injuries, utilising 4.5 mm cancellous screws and 1.6 mm K-wires. The outcome of function and anatomical reduction were assessed by AOFAS (American Orthopedic Foot and Ankle Society) midfoot score and radiography. Patients with only ligamentous injury scored poorer partly due to delay in diagnosis and the nature of the injury. The technique used maintained anatomical reduction at the time of review. PMID- 16250284 TI - Laser versus dissection technique of tonsillectomy. AB - Tonsillectomy is the single most common operation performed in Ear Nose and Throat Department. Various methods of tonsillectomy have been practiced over the century aimed at reducing or eliminating intraoperative and postoperative morbidity. Due to various blood supplies received, intraoperative bleeding is the most difficult problem and securing it is time-consuming. The time taken to control the bleeding would invariably determine the length of operation. Common postoperative complications are bleeding and pain. This study evaluated the operative time, intraoperative blood loss, postoperative pain and other postoperative complications of tonsillectomy performed by laser as compared to conventional dissection technique. This is a prospective randomized study whereby sixty patients were divided into two groups of equal number. In one group, the tonsillectomy performed by laser and in the other group the tonsillectomy performed by conventional dissection technique. Operative time and amount of blood loss is significantly reduced in the laser group. Total postoperative pain and post operative complications were not significantly different between the two groups. Tonsillectomy by using laser have shown less intraoperative bleeding and shortened the operative time. In the hospital where laser machine and expertise are available, it is justifiable to use this technique as effective method of performing tonsillectomy. PMID- 16250285 TI - Adhesive small bowel obstruction (ASBO) in children--role of conservative management. AB - Adhesive small bowel obstruction (ASBO) is an annoying postoperative complication. Though the diagnosis can be made easily, the role of conservative management in children is controversial. Hence a study was conducted to determine the role of conservative management, and to identify the factors that can predict / influence the outcome of conservative treatment in children with ASBO. Children admitted with ASBO from 1980 to 2002 (22 year period) formed the material for this study. The data was analyzed with respect to the influence of age at the time of presentation, primary disease for which original laparotomy was done, time interval between the primary surgery and the development of ASBO and the number of laparotomies prior to the development of ASBO on the outcome of conservative management. There were 74 episodes of ASBO in 69 children (Five children had two episodes). Out of 74 episodes, 5 episodes (6.75%) needed immediate laparotomy for suspected gangrene. All others were managed conservatively. Of the 69 episodes managed conservatively, 36 responded to conservative treatment (2-5 days) while 33 required subsequent surgical intervention, with 11 of them requiring bowel resection (two for gangrene and 9 for bowel damage during adhesiolysis) and in the rest 22 cases adhesiolysis. A substantial number of children with ASBO respond well to conservative treatment. Majority of the children developed ASBO within three months after the primary laparotomy. Children below the age of one year (at the time of presentation with ASBO) responded poorly to the conservative management. Children who had primary surgery for Hirschsprung's disease and intussusception also appeared to have responded poorly to conservative management, but statistically not significant. Time interval between the primary surgery and the number of laparotomies before the child developed ASBO did not influence the outcome of conservative management. PMID- 16250286 TI - Cotton bud and ear cleaning--a loose tip cotton bud? AB - Complications that arise from self-cleaning the external ear canal are common. This is a prospective study using standard questionnaire on the practice of ear cleaning in 50 subjects. A loose tip cotton bud was also shown to them to assess if it was acceptable to replace their current cotton bud. Thirty six percent of them clean their ears by introducing an object into the ear canal once or more a day. The commonest reason to clean the ears was the presence of earwax. Only 6% used a cotton bud to mop fluid from the ear canal. Almost all (92%) the subjects used a cotton bud to clean the ear. A complication rate of 2% was noted. A loose tip cotton swab was shown to the subjects. Despite explaining that it is safer and better, only 24% of the subject were willing to change to this loose tip cotton bud. Seventy four percent of the subjects cleaned their ear regularly because of earwax. The misconception of needing to clean the ear canal by introducing an object into the ear is rampant. This practice should be avoided and health care providers can play an important role in this. PMID- 16250287 TI - Pitfall in diagnostic trauma laparoscopy. AB - The resurging interest in diagnostic laparoscopy has witnessed its increasing application in trauma surgery. Such unbridled enthusiasm has at times overlooked its shortcoming in the diagnosis and management of certain in abdominal injuries. We report and discuss one such conspicuous limitation and advocate that the use laparoscopy in abdominal trauma should be tempered with caution. PMID- 16250288 TI - Acute respiratory distress syndrome due to overdose desferrioxamine: report of a child. AB - In this article, we present an 18-month-old girl with acute iron poisoning who died from acute respiratory distress syndrome due to overdose of desferrioxamine. Our purpose is to emphasize the importance of close follow-up children with acute iron poisoning for desferrioxamine toxicity. PMID- 16250289 TI - Case report of a difficult venous access: retroperitoneal approach of inserting an improved non-heparinised port for long-term use. AB - This is a case report of a patient with difficult venous access following thrombosis of major superficial and deep veins of the limbs as documented by ultrasound doppler and venography. The insertion of a few central ports were infected and the vein thrombosed. Venography revealed that central venous access was no longer feasible. The previous laparotomies had resulted in dense intra peritoneal adhesions, and rendered further laparotomy virtually impossible. The patient had occasional adhesion colics, vomiting and hypoglycemic episodes. A rarely performed retro-peritoneal approach of inserting an improved non heparinised port proved to be effective for long-term management of this patient. The surgical approach and the selected port are discussed. PMID- 16250290 TI - Metastatic cervical carcinoma in the caecum. AB - A patient who presented with acute intestinal obstruction had a right hemicolectomy for a caecal tumour. The histopathology report confirmed metastatic carcinoma in the caecum from the cervical carcinoma. Caecum is a very rare site of metastasis from cervical carcinoma. From our literature review, there have been no such cases reported. PMID- 16250291 TI - Neonatal meliodosis: very rare but be aware. AB - Meliodosis is an infectious disease encountered mainly in tropics. It is not an uncommon problem in Malaysia especially in areas with agricultural activities. Although it can occur in all age groups, there have been few reported cases in children. Men are more commonly affected than women due to outdoor activities. Neonatal cases have been reported in Hawaii and Thailand. These infants presented with neonatal sepsis or meningitis. The mode of transmission to these infants has not been elucidated. This is the report of such a case first reported in Hospital Tengku Ampun Afzan, Kuantan. PMID- 16250292 TI - Nasal cylindrical cell papilloma. AB - Inverted papilloma is the most common benign tumor of nose and paranasal sinuses arising from lateral nasal wall and middle meatus. Histologically these tumors are composed of epithelial nests that are inverted, exophytic and cylindrical. Here we describe a clinical case of nasal cylindrical cell papilloma, which was treated by endoscopic excision. PMID- 16250293 TI - Glomus tympanicum: a report of two cases. AB - Two cases of glomus tympanium tumor of the middle ear stage 'type A' according to Fisch classification, are presented due to their rarity in this part of the world. These tumors were excised by end aural tympanotomy approach and there has been no recurrence in both patients to date. PMID- 16250294 TI - Extended pedicle rectus abdominis myocutaneous flap for thigh reconstruction. AB - A rectus abdominis myocutaneous flap can provide a large amount of tissue for defect coverage. Rarely a flabby and redundant abdominal tissue was used as a huge extended flap. We report a case of recurrence malignant fibrous histiocytoma of the thigh which was radically resected. The resultant massive defect was success reconstructed with an extended pedicle inferiorly based rectus abdominis myocutaneous flap. PMID- 16250295 TI - Automatic top loader washing machine related injury. A report of four cases with serious injury. AB - We are reporting four cases of serious washing machine related injury that presented within a period of 5 months. All patients were young children with the mean age of 9 year-old and three had their dominant hand injured. The washing machines involved were the automatic top loader type and all injuries occurred during the spinning phase. Serious automatic washing machine injury is not uncommon in Malaysia. We feel that there is a need to improve the safety features especially during the spinning phase. The operating instructions and safety precautions on the washing machine should be displayed in different languages that can be understood well. Parents should also aware of the potential risks of this seemingly benign household appliance. PMID- 16250296 TI - Lingual thyroid--a lesson to learn. PMID- 16250297 TI - Easy lens cleaning solution for laboratory microscopes. PMID- 16250298 TI - Farmers and formulations--rural health perspective. AB - Conventional farming demands excessive use of chemicals in the form of synthetic fertilizers and pesticides, confirming to the norms of Green Revolution. Farmers in general, specifically in the developing countries resort to injudicious and excessive use of pesticides which is linked to the illiteracy and poverty of the rural farming community. Their overriding concern for profitable agriculture, has rendered the health of the farmers at a greater risk of developing dreadful maladies including various type of cancers, reproductive disorders, respiratory, dermal, and neuropsychological problems etc. The possible means of reducing the health risks are discussed, including the global effort to regulate the manufacture, transport and use of highly toxic pesticides. Slow and programmed transition to alternative agriculture and strengthening of farmers' knowledge on health, ecosystem and environment will prove effective. PMID- 16250299 TI - Opportunities for new research using the post-2001 ONS Longitudinal Study. AB - The addition of 2001 Census data to the ONS Longitudinal Study extends the range of research topics that this unique data resource can support. Census questions on religion, care-giving and self-rated health that were asked for the first time in 2001 and the repetition of 1991 questions on limiting long-standing illness and ethnicity raise opportunities for new longitudinal investigation in these areas. This article describes how new 2001 methodologies including data imputation, the One Number Census and de jure enumeration affect the LS database. The support service for existing and prospective LS users is described. PMID- 16250300 TI - The long shadow of childhood: associations between parental social class and own social class, educational attainment and timing of first birth; results from the ONS Longitudinal Study. AB - In September 2004, linked data from the 2001 Census was made available in the Office for National Statistics (ONS) Longitudinal Study (a 1 per cent sample of the population of England and Wales). The study now includes information from four censuses on sample members and the people they lived with. The availability of this new information, the length of follow-up and other features of the study (such as records of births and deaths to sample members) provide new opportunities for analysing change--both over the life course, between time periods, and between generations. This article illustrates the potential for analysis of continuity and change with new results on intergenerational social mobility, and on parental social class and age at first birth. PMID- 16250301 TI - First births by age and education in Britain, France and Norway. AB - Progressively later starting of childbearing has been a feature of cohort change in fertility across Europe and elsewhere over recent decades. Growing differences in the age patterns of childbearing between the Anglo-American and continental European countries, however, have also been found. The present study uses large linked-record databases in Britain, France and Norway to analyse these differences in more detail, focussing on age at entry to motherhood (first childbearing) by level of educational attainment among women born in the 1950s and in the 1960s. The shift between these two cohorts towards a later pattern of first childbearing in Britain was confined to women with secondary school qualifications and above. For women born in the 1960s, the peak age for risk of first childbearing among those with secondary school qualifications grew to be between seven and eleven years later than among women without secondary school qualifications. In France and Norway, the peak ages for risk of first childbearing shifted more uniformly across education levels between the two cohorts. For these 1950s and 1960s cohorts, improvements in women's educational levels also occurred more uniformly in France and Norway, moving more women into education categories characterised by later patterns of first childbearing. PMID- 16250302 TI - Stability and change in ethnic groups in England and Wales. AB - The Office for National Statistics (ONS) Longitudinal Study (LS) is an exceptional resource for exploring dynamic processes in individuals' lives for a representative sample of the population of England and Wales and across a thirty year period, including how those processes vary by ethnic group. However, analyses tend to assume a certain stability in the meaning of the ethnic group being studied: the insights into ethnic group differentiation are premised on the fact that the group has the same meaning over time. Here we show how the LS allows us to challenge such notions of group stability. This has practical implications for the ways we measure and conceive of Britain's minority ethnic groups. We illustrate this point with two examples: one exploring the change in ethnic group identification by the same individuals between 1991 and 2001, and the second exploring how loss to follow up is differentially experienced according to ethnic group. We provide some suggestions on the implications of this ethnic group instability for other research. PMID- 16250303 TI - Overcoming difficulties in implementing a universal newborn hearing screening program. AB - The aim of this paper was to present our experience with a universal newborn hearing screening program, including the specific problems and difficulties faced since its beginning, along with the measures used to overcome them and to improve its efficiency. We analyzed data from 22,195 newborns screened by transiently evoked otoacoustic emissions (TEOAE) performed during the first days after birth. 84.8% of the newborns passed on the first test and another 12.15% passed on rescreening before hospital discharge. This produced a "not pass" rate (false positive and true positive) of 3.05%. The rate of newborns who did not undergo screening and the rate of "lost to follow-up" newborns were reduced in time, due to various applied modifications to the protocol. It may be thus concluded that our protocol proved to be successful in attaining low refer rates for follow-up screening. A major problem that remains unresolved is the absence of effective follow-up. PMID- 16250304 TI - Identification of an ancestral haplotype of the 35delG mutation in the GJB2 (connexin 26) gene responsible for autosomal recessive non-syndromic hearing loss in families from the Eastern Black Sea Region in Turkey. AB - Mutations in the GJB2 gene have been shown to be the major cause of autosomal recessively inherited, prelingual, non-syndromic hearing loss. 35delG was found to be the most frequent mutation among Caucasians. In this study, we performed haplotype analysis of two large families with autosomal recessive non-syndromic hearing loss (totally 33 affected, 37 unaffected) from Trabzon (a city from the Eastern Black Sea region) by using polymorphic markers close to the 35delG mutation region, and identified a common haplotype, "2-6-4". The frequency of the mutant chromosomes having the 2-6-4 haplotype was compared between the Eastern Black Sea region and the other regions of Turkey and the difference was found to be significant (chi squared = 5.13/df = 1/p = 0.023). Also, when the frequency of mutant and wild type chromosomes having the 2-6-4 haplotype was compared in the Eastern Black Sea region, a statistically significant difference was observed in the mutant chromosomes (chi squared = 7.46/df = 1/p < or = 0.01). The results of this study demonstrate that the ancestral haplotype of the chromosomes bearing 35delG mutation in the Eastern Black Sea region is "2-6-4". PMID- 16250305 TI - The first camps in Turkey for asthmatic children: six years' experience. AB - The first asthma camp in Turkey was organized for one week in Iznik in September 1996. The camps were continued annually around the same time of the year in 1997, 1998, 2000, and in the consecutive years thereafter. The camp includes educational, sports and social activities. Children's knowledge about asthma and their attitudes towards physical and social activities were evaluated by a questionnaire. Pulmonary function tests were performed on the first and last day of the camp. There was no statistically significant difference between the mean spirometric values of the first and last day of the camp in spite of a vigorous physical and social program. Children enhanced their skills and knowledge about asthma and gained self-confidence in participating in sportive and social activities during the camp program. They did not experience any emergency room visit in the following year. In conclusion, a summer camping experience is very beneficial for asthmatic children in terms of both self-education and social and physical participation. PMID- 16250306 TI - Use of polymerase chain reaction for detection of adenovirus in children with or without wheezing. AB - Eighty percent of asthma attacks in children are accompanied by an upper respiratory tract viral infection. Adenovirus is one of the major viral causes of childhood bronchiolitis. As the polymerase chain reaction (PCR) is the most sensitive technique for documenting viral respiratory infections, the PCR method was performed on the throat swab samples of asthmatic children with and without wheezing to investigate the presence of the adenovirus genome in the upper respiratory tract. The frequencies of adenovirus in asymptomatic and symptomatic asthmatic patients, healthy controls and wheezy children were as follows: 33.3%, 71.4%, 37% and 62.96%, respectively. The adenovirus was detected in a significantly higher percentage in the upper airways of patients with asthma exacerbation and in children with wheezing than in patients without asthma exacerbation and in the healthy controls (p < 0.05). The frequency of adenovirus was not different between asthmatic patients receiving or not receiving inhaled corticosteroid. Adenovirus has the potential to precipitate asthma exacerbations in asthmatic patients; its frequency was not affected by the treatment of inhaled corticosteroid. PMID- 16250307 TI - Prenatal echocardiographic diagnosis of congenital heart disease: comparison of past and current results. AB - The increased experience in interpretation of fetal echocardiographic images may change the accuracy of fetal echocardiography in diagnosing fetal heart defects. We thus decided to evaluate the specificity and the sensitivity of our fetal echocardiographic examinations in diagnosing congenital heart disease, focusing especially on the outcome of complex cardiac pathologies. Between October 1999 and July 2003, 642 fetuses were followed until birth and underwent a postnatal reassessment of the cardiovascular system in our institution. These cases constitute our cohort. The postnatal reassessment was mainly done by echocardiography; some cases also had angiography. In case of intrauterine or postnatal death, an autopsy was performed. The prenatal and postnatal diagnoses were compared, and specificity and sensitivity of fetal echocardiography for congenital heart pathologies were determined. Among 45 affected pregnancies, 31 cases had complex and 14 had significant cardiac defects. The sensitivity of fetal echocardiography for cardiac anomalies was 93.3%; the specificity was 100%. Compared to our previous study, the sensitivity was remarkably improved (in our previous study sensitivity was 78% and specificity 100%). Echocardiography is a very useful and reliable tool in the evaluation of the fetal cardiovascular system, and has high sensitivity and specificity for congenital heart diseases. PMID- 16250308 TI - The efficacy of immunoglobulin replacement therapy in the long-term follow-up of the B-cell deficiencies (XLA, HIM, CVID). AB - Immunoglobulin replacement therapy is the essential treatment of B-cell deficiencies. Because of the high expense of therapy, optimal dose, infusion intervals and serum IgG levels should be well defined. Data of 19 X-linked agammaglobulinemia (XLA), 7 hyper-IgM syndrome (HIM) and 20 common variable immunodeficiency (CVID) patients were analyzed. Infection frequencies and hospitalization requirements were correlated with the immunoglobulin doses used and serum IgG levels achieved. The characteristics before diagnosis and after treatment were compared among the XLA, HIM and CVID groups. By using a median dose of 370 mg/kg/month immunoglobulin, which maintained serum IgG levels at a median concentration of 440 mg/dl, the annual incidence of infections dropped from 12.4 to 3.2 and annual hospitalization requirements decreased from 1.6 to 0.16 per patient. Serum IgG levels of 300-500 mg/dl were found to be satisfactory, except in the CVID group. Increasing the level over 500 mg/dl neither prevented pneumonia further nor decreased the need for hospitalization. Monthly replacement was found to be adequate, except for XLA patients. Serum IgG levels between 300-500 mg/dl are sufficient for effective treatment of hypogammaglobulinemias. These concentrations can be maintained with 300-400 mg/kg/month doses. Higher doses and IgG levels are not needed. PMID- 16250309 TI - Spontaneous closure of small apical muscular ventricular septal defects. AB - Between 1993 and 2002, apical muscular ventricular septal defects were identified in 99 cases by echocardiographic examination. Spontaneous closure time was analyzed in 42 cases followed up at least two years, retrospectively. The ages of 42 cases ranged between 1 day and 13 years old. Initial examination was performed in 22 cases within the newborn period, in 8 cases between 1 and 6 months of age and in 12 cases at more than 6 months of age. Spontaneous closure was seen in 24 of 42 cases (57.1%) between 1 and 36 months of age, and it was most commonly recorded during the first 6 months. It was remarkable that spontaneous closure occurred in 20 of 22 cases (90%) diagnosed in the neonatal period. Closure was seen in 4 of 8 cases (50%) whose initial examination was performed between 1 and 6 months and in none of the patients diagnosed at more than 6 months of age. In conclusion, the frequency of spontaneous closure in cases diagnosed in the neonatal period is higher than previously believed. It is advisable to follow up patients to determine spontaneous closure, especially within the first two years of life. PMID- 16250311 TI - Familial ureteroceles: an evidence for genetic background? AB - In the pediatric population, ureteroceles may present with different clinical pictures, and the severity of the renal damage is greater than in adults. Ureterocele, an anomaly of ureteric budding, is likely a component of a spectrum of anomalies including vesicoureteral reflux and ureteral duplications. Both have been confirmed to have a genetic and familial basis. We document the largest series of familial cases of ureteroceles, giving evidence for genetic background. We retrospectively reviewed the charts of patients with familial ureteroceles seen between 1992 and 2002. Coexisting ureteral anomalies and features of the cases were documented and compared to sporadic cases and all familial cases within the literature. This is the largest series of familial ureterocele patients in the literature. The review of the literature revealed seven publications with seven ureterocele families (15 affected patients) between 1936 and 2002. Comparing sex, ureterocele location, and single versus duplex systems, familial series are similar to other sporadic cases. Three of the families have twin siblings with ureteroceles. Familial cases, despite their rarity, raise the issue of the genetic origin of uretereoceles. Family members of ureterocele cases should be informed and followed carefully, especially twins. Increased reporting and genetic analysis of familial ureteroceles may prove to link the genetic mouse models of abnormal ureteric budding to the human conditions. PMID- 16250310 TI - Neutrophil hypersegmentation and thrombocytosis in children with iron deficiency anemia. AB - Neutrophil hypersegmentation is an expected peripheral blood smear finding in megaloblastic anemias. But some clinical reports suggest that neutrophil hypersegmentation may also occur in patients with iron deficiency anemia. In this study we searched the presence of neutrophil hypersegmentation and thrombocytosis in patients with iron deficiency anemia but who had normal serum vitamin B12 and folic acid levels. The study comprised 102 patients with iron deficiency anemia and 21 age-matched healthy controls. All routine tests for iron deficiency anemia were done, serum folate and cobalamin levels were measured, and platelets were counted in all patients and controls. Peripheral blood smears were examined for neutrophil hypersegmentation. Hypersegmentation was found in 30.4% of anemic patients and in 9.5% of controls (p < 0.05). The number of platelets was also significantly higher in anemic children (p < 0.05). These results show that neutrophil hypersegmentation may also be seen in patients with iron deficiency anemia, and thrombocytosis is a common laboratory finding in this disorder. PMID- 16250312 TI - The contribution to success of various methods of treatment of temporomandibular joint ankylosis (a statistical study containing 24 cases). AB - Temporomandibular joint (TMJ) ankylosis is an important joint disorder which, in addition to emerging through trauma, can also arise as a result of local and systemic infections. TMJ ankylosis which develops in childhood in particular has its own characteristics. Despite the existence of different views on the treatment of TMJ ankylosis, various techniques have been defined, and three basic techniques are currently employed: gap arthroplasty, interpositional arthroplasty and joint reconstruction. Our study is intended as a statistical inquiry into the contribution to the operation success of the three different methods of treatment. Moreover, the effect on treatment outcome of unilateral or bilateral ankylosis is also statistically evaluated. In our study, eight cases were treated with gap arthroplasty, nine with interpositional arthroplasty and seven with joint reconstruction performed by costochondral graft. As a result of our statistical evaluation, it was determined that the effect of interpositional arthroplasty on post-operative maximal interincisal mouth opening was greater than that observed with the other methods. PMID- 16250313 TI - A very rare cause of recurrent apnea: congenital nasopharyngeal teratoma. AB - A three-month-old girl patient born at the 37th week of gestation as one of twins was admitted to our hospital with cardiac arrest. There was past medical history of multiple hospitalizations with symptoms of cyanosis, respiratory distress, and frequent and severe attacks of apnea since her birth. On nasopharyngeal computerized tomography a mass (25x24 mm) occupying the right side of the nasopharynx was detected. The pathological evaluation of the excised mass revealed benign teratoma. After the removal of the nasopharyngeal mass under general anesthesia, respiratory distress and attacks of apnea disappeared completely. In every neonate with unexplained stridor and recurrent apnea, beside the common causes like respiratory distress syndrome, sepsis, and asphyxia, the possibility of oropharyngeal and nasopharyngeal mass should be kept in mind. PMID- 16250314 TI - Autoimmune thrombocytopenic purpura after mumps infection. AB - Autoimmune thrombocytopenic purpura is estimated to be one of the most common acquired bleeding disorders of children. The pathogenesis involves the generation of autoantibodies against the normally expressed glycoproteins on the platelet membranes. These antibody-coated platelets in turn are destroyed by the spleen and other reticuloendothelial organs. Although the disease can occur without an identifiable etiology, many underlying pathologies, including infections, can be found. We report the mumps virus as a rare etiology of secondary autoimmune thrombocytopenic purpura. PMID- 16250315 TI - Brucella melitensis in blood cultures of two newborns due to exchange transfusion. AB - Brucellosis is a zoonotic infection, transmitted to humans primarily by consumption of unpasteurized milk and milk products. Transmission by blood transfusion is possible but very unusual. Herewith we present two newborns with positive blood cultures for Brucella melitensis after exchange transfusions. However, the standard tube agglutination titers against Brucella were not elevated and the newborns did not develop brucellosis. It is suggested that, in areas endemic for brucellosis, blood donors should be questioned about symptoms of brucellosis, and if suspected, serological tests for brucellosis should be indicated before blood transfusion. At the same time, the prevalence of the disease among animals should be reduced with effective animal disease control programs. PMID- 16250316 TI - Startle disease-two sibling cases. AB - Startle disease (hyperekplexia) is a rare non-epileptic disorder characterized by hypertonia, generalized stiffness and brief muscle jerks in response to unexpected auditory, somatosensory and visual stimuli. In this paper, two siblings with generalized stiffness and sudden muscle jerks to unexpected stimuli of various types are presented. They were previously misdiagnosed as epilepsy and treated with the conventional antiepileptic drugs. We wanted to call attention to and increase awareness of this rare disease and its differential diagnosis to avoid unnecessary investigations and treatment. PMID- 16250317 TI - Catheter-associated recurrent intracardiac thrombosis and factor V Leiden mutation in a child with non-Hodgkin's lymphoma. AB - Patients with cancer have an increased risk for thromboembolism, which might be related to several factors including central venous catheters and chemotherapeutics. Congenital prothrombotic risk factors might also contribute to thrombotic events. In this report, we present a catheter-related recurrent intracardiac thrombosis in a boy with non-Hodgkin's lymphoma and factor V Leiden mutation. Screening for factor V Leiden mutation in children with cancer and recurrent thrombotic events is recommended. Periodic echocardiography may be considered for a group of patients if the catheter tip is in the right atrium and therapy includes L-asparaginase and corticosteroids. PMID- 16250318 TI - Spontaneous rupture of choledochal cyst presenting in childhood. AB - Spontaneous rupture of choledochal cysts is one of the rare complications, and can sometimes be the initial manifestation. It should be considered in the presence of bile-like fluid. A 10-year-old girl had acute onset of abdominal pain, vomiting, and elevated bilirubin, alkaline phosphatase, glutamic oxaloacetic transaminase and glutamic-pyruvic transaminase levels. There was no trauma in her history. In ultrasonography and computed tomography, dilated common bile duct, cystic mass of 10 x 6 cm, and free intraperitoneal fluid in abdominal cavity were demonstrated. Radiological methods, especially intraoperative cholangiography, should be performed for evaluation. We report a case of spontaneous rupture of the choledochal cyst with clinical and radiological findings. PMID- 16250319 TI - Brain calcification due to secondary hyperparathyroidism in a child with chronic renal failure. AB - Secondary hyperparathyroidism (SHPT) has been better treated over the last decades, but the rate of metastatic calcifications, which were rarely seen before, was significantly increased in dialysis patients. The presence of uncontrolled SHPT, disorders of calcium (Ca) and phosphorus homeostasis and the common usage of large doses of vitamin D and Ca- containing phosphate binders may all contribute to the metastatic calcifications of soft tissues and vasculature leading to some life-threatening complications. Although the metastatic lung, heart, kidney, intestinal wall, skin, eye and soft tissue calcifications have been commonly reported in adults and also in children undergoing dialysis, the central nervous system calcification is a very rare condition. We report here a pediatric hemodialysis patient who presented with severe neurological findings due to the metastatic brain calcification secondary to his uncontrolled hyperparathyroidism. PMID- 16250320 TI - A case of double-outlet left ventricle with atrioventricular discordance. AB - The clinical, echocardiographic and angiographic aspects of a four-month-old boy with double-outlet left ventricle, atrioventricular discordance, L-malposition of the great arteries, ventricular septal defect and pulmonary stenosis are described. Additionally, in this patient, the right arcus aorta and the ligament of ductus arteriosus caused anterior compression of the trachea. The anterior position of the morphological left ventricle is the most interesting feature. PMID- 16250321 TI - Unilateral pulmonary agenesis associated with colloidal goiter in a newborn: a case report. AB - Unilateral pulmonary agenesis is a very rare developmental malformation that is often associated with other anomalies. It can be asymptomatic or present with respiratory symptoms. Our case is a female newborn infant who had been taken to the hospital suffering from difficulty in breathing at the first day of birth. The baby died at the age of three days due to respiratory failure. On autopsy examination and its histopathological evaluation, we detected right pulmonary agenesis and colloidal goiter. According to the literature, pulmonary agenesis is associated with other anomalies including esophageal atresia, tracheal stenosis, musculoskeletal anomalies, DiGeorge syndrome and cardiovascular malformations such as septal defects, patent ductus arteriosus and total anomalous pulmonary venous return. To our knowledge, this is the first case of pulmonary agenesis associated with colloidal goiter. PMID- 16250322 TI - The triad of nesidioblastosis, congenital neuroblastoma and glomerulocystic disease of the newborn: a case report. AB - Neuroblastoma is the most common malignant tumor of the newborn, comprising 20% of all malignancies encountered during the neonatal period. We herein report a newborn who was born after 29 weeks' gestation and died unexpectedly at the 12th hour of life with no response to vigorous cardiopulmonary resuscitation. Autopsy findings revealed a right pararenal mass; microscopic examination showed neuroblastoma. Although the pancreas was grossly normal, its microscopic sections revealed a reduced number of islets of Langerhans and dispersion of the islet cells throughout the exocrine cells of the pancreas, and immunocytochemistry for the pancreatic hormones confirmed the dispersion of the islet cells. Final pathologic interpretation thus concluded the presence of nesidioblastosis. Furthermore, microscopic examination of the kidney showed glomerulocystic disease. Although the association of congenital neuroblastoma and nesidioblastosis has recently been defined as a new complex, neurocristopathy, the triad of congenital neuroblastoma, nesidioblastosis and glomerulocystic disease of the newborn has not been reported previously. To our knowledge, our case is the first reported newborn presenting with this triad. In conclusion, the association of nesidioblastosis and/or renal glomerulocystic disease should be kept in mind when encountering a case of congenital neuroblastoma. However, whether the presence of glomerulocystic disease in association with those other neurocristopathic pathologies is a coincidental finding or shares a common pathophysiological mechanism remains to be determined. PMID- 16250323 TI - [Medicodemographic problems in Russian Federation]. AB - The authors of the article analyze the current health status of Russian population and underline the necessity to set priorities in the development of the country's public health system. Within a short period of time the mortality in this country has risen 1.5 times, and the birth rate have declined twice. By January 1, 2005, the population of Russian Federation had decreased by 10,361,300 people, as a result of natural depopulation. Mostly affected by the crisis is able-bodied population, the mortality among who have increased 2.5 times. The gap in the mortality between ours and other economically developed countries is now sevenfold. This means that six out of seven cases of death at the productive age in this country are absolutely preventable. The mortality among pensioners has not changed at all, i.e. people at this age seem to be reliably protected against the crisis. The birth rate has decreased abruptly; its summary coefficient has fallen down to 1.17 while 2.15 are necessary for simple reproduction. The proportion of out-of-wedlock children, as well as that of disabled children, displays a multiple increase. Male population in Russia demonstrates a catastrophic super mortality, the gap between the length of life among men and women being 13.5 years. In a number of regions men do not reach the age of 54. PMID- 16250324 TI - [Molecular and genetic study of the role of hormones, receptors, and enzymes in regulation of reproduction, lipid metabolism, and other human physiological functions]. AB - Prevalence of uterine progesterone receptors over estrogen ones, high uterine cAMP level, and low uterine prostaglandin level are necessary conditions of normal pregnancy. In cases of spontaneous and antiprogestin RU486-induced abortions, estrogen receptors prevail over progesterone ones, cAMP level decreases, and prostaglandin concentration in decidual tissue increases. Porcine and bovine beta-lipotropines were the first proteins, whose correct amino acid sequence was first determined in Russia. Several research centers carried out collaborative studies of the nucleotide sequences of human and animal proopiomelanocortin (lipotropin precursor) and prolactin cDNA. Researchers constructed genetic engineering producers of human pre-proinsulin and somatostatin, identified structural genes expressed in pancreatic beta-cells, studied antigenic properties of glutamic acid decarboxylase (GAD), which determine insulin-dependent diabetes, and identified the cholesterase determinant. They revealed mutations in the genes of proopiomelanocortin and melanocortin receptors (MC4-P), which inhibit leptin regulation of appetite and are associated with human obesity. PMID- 16250325 TI - [Mitochondria as the target for neuroprotectors]. AB - Contemporary methods of treatment of neurodegenerative diseases (NDs) are limited and mainly symptomatic. Despite difference in particular clinical manifestations, NDs have a number of common features, the main of which is death of certain neuron population. The authors suppose that mitochondria and the phenomenon of mitochondrial permeability transition (MPT), playing the key role in cell death, may be a perspective target for the search of drugs with a capability of delaying the neurological deficit associated with NDs. The authors have demonstrated that some neurotoxins which are widely used to model neurodegenerative conditions are able to potentiate or even induce MPT. The neuroprotective effect of widely used cognition-enhancing drugs, such as tacrin, memantine, dimebon, N acethylserotonine, and extract of Gingko biloba), may also be a result of their interaction with mitochondria. Thus, the search of drugs capable of preventing or breaking the cascade of cell death as a result of MPT suppression and, at the same time, compensating for the impaired brain functions, is very topical. PMID- 16250326 TI - [Immune and interferon status of primates]. AB - The authors of the article have first determined standards of interferon (IFN) status of healthy primates (M. mullatta) belonging to different age groups, such as serum IFN, spontaneous IFN production, IFN-a and IFN-y production. They showed that old healthy primates have low ability of IFN-a and IFN-y production and high serum IFN level. The parameters of IFN status were compared to the number of B- and T-lymphocytes and their subpopulations; qualitative evaluation of protective properties of young and old healthy primates' immunity was carried out. PMID- 16250327 TI - [Intracavitary laser therapy and its effect on the kallikrein-kinin system of blood in children with pneumothorax]. AB - The article presents the results of low-intensity laser application in complex treatment of 137 children with acute purulent destructive pneumonia complicated by pneumothorax with bronchial fistulas. A method of intracavitary laser therapy, developed in the clinic, allowed obliteration of bronchopleural fistulas without application of bronchial occlusion and other invasive techniques. Evaluation of the kallikrein-kinin system of blood revealed prominent reduction of kininogenesis in most (87%) patients upon admission (3 weeks after the onset of the disease), which is an important link of the pathogenesis of late stages of complicated acute purulent lung destruction in children. The study also demonstrated that low-intensity laser emission modulates pyoinflammatory process due to its effect on cell-mediated immunity, neutrophilic phagocytosis and the kallikrein-kinin system of blood. Intracavitary laser therapy is the treatment of choice in children with acute purulent destructive pneumonia complicated by pneumothorax with bronchial fistulas. Application of intracavitary laser therapy in complex therapy of complicated acute purulent lung destruction in children allowed discharge from the hospital 5 to 7 day earlier, and prevented lung inflammatory process chronization. None of the patients have died within last 10 years. PMID- 16250329 TI - [The anticancer drug Kang-Lai-Te emulsion for infusion]. AB - Kanglaite (KLT) emulsion for infusion is a new type of anticancer drug, prepared by extracting active antitumor components from the primary product of the Chinese plant Semen Coicis using modern technology, and formed as lipid emulsion for intravenous and intra-arterial injections. Clinical application of this drug demonstrates high efficacy of KLT in treatment of various tumors, such as lung, hepatic, stomach, and breast carcinomas. Its use leads to a significant increase of immune functions and improves life quality: when combined with radio-, chemotherapy, and auxiliary therapy, it leads to a significant increase of the therapeutic effect and reduces the toxic effects of these treatments. Deep study of the mechanism of KLT action, performed in large research centers of China, has demonstrated that the drug blocks tumor cell mitosis at the boundary of G2 and M phases of the cell cycle, induces tumor cell apoptosis, increases the expression of Fas/Apo-1 gene, which inhibits the growth of tumor cells, and reduces the expression of Bel-2 gene, which promotes it, inhibits angiogenesis, actively decreases cancer cachexy, and is able to overcome multiple drug resistance of tumor cells. PMID- 16250328 TI - [Positron emission tomography in oncourology]. AB - The authors present the results of the examination of 61 patients with genitourinary space-occupying lesions, using 18F- fluorodeoxyglucose positron emission tomography (PET) in whole body mode. In all cases the diagnosis was verified morphologically. The results demonstrated high diagnostic accuracy of PET, including possibility to determine the extent of oncourological cancer. However, the method displays poor efficacy in cases of hypernephroid cancer due to low level of glycolysis in this type of tumor. PMID- 16250330 TI - [Achievements and prospects of clinical abzymology]. AB - Catalytic autoantibodies (abzymes) are autoantibodies that are potentially ready to realize certain effects in the organism, first of all antibody-mediated catalysis and cytotoxicity. Natural abzymes with protolytic (protabzymes) and DNA hydrolyzing DNA-abzymes) activity are of the greatest interest. The most impressive example of the catalytic activity of protabzymes is hydrolysis of specific proteins, revealed in patients with autoimmune diseases, such as bronchial asthma (vasoactive intestinal neuropeptide), autoimmune thyroiditis (thyroglobulin), multiple sclerosis (myelin basic protein), and autoimmune myocarditis (cardiomyosin). The pathogenic role of DNA-abzymes is not quite clear yet. However, it has been proven that they present a powerful regulator of apoptosis and other cytotoxicity mechanisms in systemic autoimmune diseases and tumors. The most promising is use of abzymes as illness activity markers, and as therapeutic agents capable of catalyzing specific proteins or activating antitumoral chemotherapeutic preparations. PMID- 16250331 TI - [Associated synthesis of some secreted pathogenicity factors of Klebsiella pneumoniae]. AB - The study was carried out to evaluate the substrate specificity and activity of proteases secreted by strains of Klebsiella pneumoniae with various degree of virulence. The process included cultivation of the strains in semi-synthetic medium, after which the biomass was inactivated and the supernatant was separated from bacterial cells through centrifugation. Elastase-, trypsin-, and chymotrypsin-like proteolytic activity was measured in the supernatant and in all fractions obtained through gel-filtration, followed by DEAE-sepharose purification. Regardless of the degree of virulence, all the studied strains of K. pneumoniae secreted only one proteolitic enzyme, which was elastase with molecular weight of about 21 kDa. Addition of glycoprotein--the main structural component of eucaryotic cells--into the culture medium in the beginning of incubation, increased protein, polysaccharide, and lipopolysaccharide synthesis; proteolythic activity in the supernatant fluid increased from 7,476 to 15,731 mU/ml. The increase was associated with an elevation of polysaccharide synthesis from 173 to 349 mg dry weight. However, proteolythic activity per 1 gr of polysaccharide did not increase; it was 43.3 and 45.1 units, respectively. Thus, proteolytic activity increased in direct propotion to the increase of polysaccharide synthesis into the culture medium. PMID- 16250332 TI - Molecular dynamics of the local anesthetic tetracaine in phospholipid vesicles. AB - Upon introduction into phosphatidylcholine vesicles, the 13C magnetic resonance peaks of the aromatic resonances of tetracaine are broadened while the T1 relaxation times show little change. Addition of tetracaine to vesicles containing 30% cholesterol produces a similar broadening in the 13C NMR spectrum of tetracaine. Nuclear magnetic resonance parameters of phosphatidylcholine in vesicles which are unchanged by the addition of equimolar tetracaine include 13C T1 relaxation time and 31P linewidth, T1 relaxation time, and nuclear Overhauser effect enhancement. These results are interpreted as indicating a hydrophobic interaction between hydrocarbon portions of the anesthetic and phospholipid bilayer. The rotational correlation time of tetracaine about its long axis in the vesicles has been calculated from the 13C NMR spin lattice relaxation times to be about 10(-10.3) s and is unchanged by incorporation into the phospholipid bilayer. The positively charged ammonium group of tetracaine interacts with the negatively charged phosphate group of the vesicle lipids. Using shift reagents and 31P NMR, tetracaine has been shown to displace cations from the bilayer surface, and does not undergo fast flip-flop across the vesicle bilayer. PMID- 16250333 TI - Effect of solution non-ideality on erythrocyte volume regulation. AB - A non-ideal, hydrated, non-dilute pseudo-binary salt-protein-water solution model of the erythrocyte intracellular solution is presented to describe the osmotic behavior of human erythrocytes. Existing experimental activity data for salts and proteins in aqueous solutions are used to formulate van Laar type expressions for the solvent and solute activity coefficients. Reasonable estimates can therefore be made of the non-ideality of the erythrocyte intracellular solution over a wide range of osmolalities. Solution non-ideality is shown to affect significantly the degree of solute polarization within the erythrocyte intracellular solution during freezing. However, the non-ideality has very little effect upon the amount of water retained within erythrocytes cooled at sub-zero temperatures. PMID- 16250334 TI - Isolation and incorporation into lipid vesicles of a concanavalin A receptor from human erythrocytes. AB - Affinity chromatography has been used to isolate a concanavalin A receptor portion of Band 3 from humen erythrocytes in the presence of the readily dialysable detergent, dodecyltrimethylammonium bromide. Addition of phospholipids to the isolated fraction and removal of detergent by dialysis leads to formation of vesicles containing the receptor. Intramembranous particles similar in size and shape to those seen in intact erythrocytes are a characteristic of the reconstituted preparations. Vesicles containing receptor bind concanavalin A with high affinity. PMID- 16250335 TI - Arrangement of phosphatidylserine and phosphatidylethanolamine in the erythrocyte membrane. AB - Cross-linking of phosphatidylethanolamine and phosphatidylserine in the erythrocyte membrane with the reagent difluorodinitrobenzene was studied as a function of temperature, time and concentration of difluorodinitrobenzene. The optimal extent of cross-linking of phosphatidylethanolamine to phosphatidylethanolamine, phosphatidylethanolamine to phosphatidylserine and phosphatidylserine to phosphatidylserine was expressed as molar ratios of these three different cross-linked species. The experimental results were compared to different models of a phospholipid monolayer containing phosphatidylethanolamine and phosphatidylserine in which phosphatidylserine was arranged primarily as singles (having 6 phosphatidylethanolamine neighbors) as clusters of dimers, trimer and tetramers or as large clusters. In the various model monolayers each lipid component has 6 neighbors. The models which are consistent with the experimental results are those in which phosphatidylserine and phosphatidylethanolamine occur as small clusters in a non-random array. PMID- 16250336 TI - Effects of anions, pH and magnesium on calcium accumulation and release by sarcoplasmic reticulum vesicles. AB - In the absence of oxalate, Ca2+ accumulation by isolated sarcoplasmic reticulum vesicles may show a transient behavior in which the vesicles accumulate during the first 2 min of incubation as much as twice the amount of Ca2+ which is retained after 5-7 min, when Ca2+ accumulation approaches a steady state. Before Ca2+ release begins, the Ca2+ accumulation can reach 200-250 nmol/mg protein. The spontaneous release of the "extra" Ca2+ initially accumulated appears to be triggered by the attainment of a sufficiently high concentration of free Ca2+ inside the vesicles. The amplitude of the transient phase of Ca2+ accumulation reaches a high value near pH 6.0 and is increased by free Mg2+. At optimal concentrations of H+ and Mg2+, the amount of Ca2+ accumulated during the transient is augmented by various anions, in the order maleate > or = propionate > or = succinate > chloride > sulfate > acetylglycine. The divalent anions have their maximum effects at 20-40 mM and the monovalent anions, at 40-200 mM. At 200 mM, all of the carboxylic anions tested significantly reduce the amount of Ca2+ retained in the steady state. PMID- 16250337 TI - The involvement of the plasma membrane in the development of Dictyostelium discoideum. I. Purification of the plasma membrane. AB - A method for the isolation and purification of plasma membranes of Dictyostelium discoideum by equilibrium centrifugation on sucrose followed by Renografin continuous density gradients has been developed and monitored both with electron microscopy and a number of enzyme assays. On electron microscopy, the final plasma membrane fractions are judged to be freethe basis of of nuclei, rough endoplasmic reticulum, lysosomes and peroxisomes. Some profiles of the mitochondrial inner membranes are found within the plasma membrane fractions, but this contamination has been estimated to be only 5%. On the basis on enzyme assays, the plasma membrane fractions contain all the 5'-nucleotidase activity in the final gradients and are free of catalase, acid phosphatase and malate dehydrogenase activity (markers for peroxisomes, lysosomes, soluble enzymes and the matrix of mitochondria). Their content of glucose-6-phosphatase is reduced by more than 70%. The large majority of RNA and DNA have been removed from the preparation. PMID- 16250338 TI - The role of the plasma membrane in the development of Dictyostelium discoideum. II. Developmental and topographic analysis of polypeptide and glycoprotein composition. AB - Previous workers have shown in a variety of ways that cell contact is required for the differentiation of Dictyostelium discoideum. Because interactions between cells are probably mediated by molecules on their plasma membranes, we have characterized the polypeptide composition of the membrane of cells at different stages of development. At least 55 polypeptides are found in the plasma membrane of vegetative cells. The polypeptide composition of the plasma membranes changes considerably during development. Treatment of intact cells with pronase indicated that many of the altered components appear to be located on the external surface of the plasma membrane where they could participate in interactions between cells. Similar digestion of the isolated membranes destroys most of their polypeptides, indicating that the bulk of the proteins of the plasma membrane are not completely embedded in the membrane. Several polypeptides appear to change in sensitivity to pronase during development. There are several changes in glycoprotein composition which occur between log phase and aggregation phase. An almost complete change in glycoprotein species occurs between aggregation and pre culmination. Unlike the polypeptides, the glycoproteins are very resistant to pronase treatment in intact cells. However, some are pronase sensitive in isolated membranes. PMID- 16250339 TI - On the quantitative interpretation of biomembrane structure by Raman spectroscopy. AB - The now well-established use of Raman spectroscopy to examine the structure of biomembranes is extended through an examination of the origins of the structure sensitive features of phospholipid spectra and the development of quantitative order-parameters. One parameter gives a quantitative measure of the fraction of all-trans bonds in the hydrocarbon chains while the other provides a semiquantitative estimate of the lateral crystal-like order between the chains. The parameters are used to study the differences between vesicles and dispersions of dipalmitoyl phosphotidylcholine, dimyristoylcholine and egg lecithin. We find that the vesicles of dipalmitoyl phosphotidylcholine are substantially less ordered than the dispersions in terms of both longitudinal and lateral order which are greatly decreased. A very careful measurement of the order as a function of temperature shows that there is a pre-melting transition in the dispersions of dipalmitoyl phosphotidylcholine which does not exist in the vesicles. Remarkable agreement is obtained between the Raman technique and that previously reported by calorimetric measurements and theoretical calculations. PMID- 16250341 TI - Membrane-associated components of the bacterial flagellar apparatus. AB - At the position of insertion of the flagellum into the Gram-negative bacterial cell envelope, a specialized membrane differentiation has been observed by electron microscopy. This structure, termed concentric membrane rings, is harboured on the under-side of the outer membrane of Spirillum serpens, and forms a plate-like array of up to seven rings (diameter 90 nm) and an interior supporting collar. The concentric membrane rings are sensitive to proteolytic digestion, but are lysozyme and phospholipase resistant. The structures are disrupted by ionic detergents, yet resistant to the action of non-ionic detergents. A model integrating the basal organelle of the bacterial flagellum and the outer membrane of the cell wall is presented. PMID- 16250340 TI - Purification and characterization of spiralin, the main protein of the Spiroplasma citri membrane. AB - The membrane proteins from Spiroplasma citri have been resolved into 16 components by SDS-polyacrylamide gel electrophoresis. By this technique it was also shown that the molecular weights of these proteins ranged from 13000 to 160 000. One of the proteins, which had an apparent molecular weight of 26 000 was the most abundant and represented more than 22% of total membrane protein. We have designated this protein spiralin. None of the proteins contained carbohydrate. Spiralin has been isolated by a procedure which involves removal of some membrane proteins with the neutral detergent Tween 20, selective solubilization of the Tween residue in DOC and fractionation of the DOC-soluble material by agarose-suspension electrophoresis. The homogeneity of spiralin was demonstrated by analytical polyacrylamide gel electrophoresis under different conditions and by crossed immunoelectrophoresis. Spiralin appeared to bind less DOC than the other membrane proteins of S. citri. This observation does not imply, however, that the binding of DOC to spiralin is weak. Spiralin was neither soluble in detergent-free buffers nor in Tween 20, which indicated that it is an intrinsic membrane protein. The amino-acid composition of spiralin was quite different from that of the membrane. Spiralin lacked methionine, histidine and tryptophan, and had a low content of glycine, leucine, tyrosine and phenylalanine, but a high content of threonine, alanine and valine. PMID- 16250342 TI - Characterization of gastric mucosal membranes. IX. Fractionation and purification of K+-ATPase-containing vesicles by zonal centrifugation and free-flow electrophoresis technique. AB - Methods are described for purification of a vesicular membrane fraction of hog gastric mucosa using differential centrifugation, density gradient separation on zonal rotors and free-flow electrophoresis. As a result a fraction is obtained enriched 40-fold in terms of K(+)-ATPase and free of any other enzyme marker other than K(+)-activated p-nitrophenyl phosphatase. The 5'-nucleotidase and basal Mg(2+)-ATPase are clearly separated from the latter enzymes. Osmotic shock, Triton X-100 treatment or K+ ionophores increased the K(+)-ATPase activity in isotonic conditions, but K(+)-p-nitrophenyl phosphatase is not affected by these treatments, nor is the ATPase activity in the presence of NH4+. The results suggest that the electrophoretic fraction contains a major population of tight vesicles, whose permeability to K+ is rate limiting for the ATPase activity but not for the p-nitrophenyl phosphatase activity. It is concluded that K+ site for the ATPase is internal whereas the K+ site for the p-nitrophenyl phosphatase is external, hence, the K+ site must be mobile across the membrane. PMID- 16250343 TI - Water-soluble acetylcholine receptor from Torpedo californica. Solubilization, purification and characterization. AB - The nicotinic acetylcholine receptor from electrogenic tissue of Torpedo californica was solubilized by tryptic digestion of membrane fragments obtained from autolysed tissue, without use of detergent. The water-soluble acetylcholine receptor was purified by affinity chromatography on a cobra-toxin-Sepharose resin. The purified receptor bound 4000-6000 pmol per mg protein of alpha [125I]bungarotoxin, and toxin-binding was specifically inhibited by cholinergic ligands. Gel filtration revealed a single molecular species of Stokes radius 125 +/- 10 A and on sucrose gradient centrifugation one major peak was observed of 20 22 S. Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate and beta-mercaptoethanol revealed two major polypeptides of mol. wt. 30 000 and 48 000. PMID- 16250345 TI - Stimulation of Bacillus subtilis membrane adenosine triphosphatase by cationic bactericidal agents. AB - The environmental Mg2+ used in preparation of Bacillus subtilis membranes was found to influence the responses of the associated ATPase to cetyltrimethylammonium bromide (CTAB). Membranes prepared using fluids containing higher Mg2+ levels exhibited lower control activity than was seen with low Mg2+ membranes. Increased environmental Mg2+ resulted in higher stimulations with lower doses of the agent. ATPase of all three membrane types was stimulated in two concentration ranges, but in the doses tested, CTAB inhibited the ATPase of only those membranes obtained using fluids containing high Mg2+ for every stage of the isolation. Sonication of membranes for 25 s at half maximum output yielded three fractions, consisting of a soluble form which was sensitive to CTAB stimulation at 25 microg/ml of assay mixture; small, 95-110 nm, vesicles, which were resistant to CTAB at 25, 75, and 150 microg/ml, and large vesicles, similar to untreated membranes both in morphology and responses to detergent. Combinations of detergent and protein (beta-lysin or arginine-rich histone) produced activity appearing to be additive when the protein level was present in a high concentration and the detergent was present at levels corresponding to the minimum influence. Mixtures of a maximally stimulating dose (75 or 100 microg/ ml) of detergent and a small amount of protein produced activities that were at least 92% or more of the expected sums of individual stimulations. Interference occurred with the following mixtures: high amounts of detergent and protein; high protein and 10 or 15 microg/ml CTAB; and beta-lysin and arginine-rich histone, both at high levels. These data are consistent with a hypothesis that the two peaks in CTAB stimulation reflect two adjacent ATPase sites, one of which is also susceptible to stimulation by cationic protein. PMID- 16250344 TI - Isolation of plasma membrane glycoprotein from bovine thymocytes. AB - Glycoprotein was isolated from a purified thymocyte membrane preparation by two methods: lithium diiodosalicylate-phenol extraction and hot 75% ethanol extraction. A higher yield of membrane sialic acid was obtained by the latter method. The preparations had similar apparent molecular weights on sodium dodecyl sulfate gel electrophoresis. Both had similar receptor activities against a panel of hemagglutinins, although the 75% ethanol extract was more active on a weight basis. However, there were significant differences in carbohydrate and amino acid compositions of the two thymocyte extracts. The lithium diiodosalicylate extracted material had much more glucose, ribose, and glycine than the ethanol extract. The glycoprotein preparations from thymocytes were quite distinct from the glycoprotein of bovine erythrocytes in both composition and receptor properties. PMID- 16250346 TI - The effect of temperature on monoxygenase reactions in the microsomal membrane. AB - The effect of temperature on the rates of monoxygenase reactions was studied with microsomes prepared from phenobarbital pretreated rats. The rates of the N demethylation of ethylmorphine, benzphethamine, aminopyrine, and p-nitroanisole were studied. Breaks at temperatures around 24 degrees C were observed in the Arrhenius plots of all these reactions. The energy of activation of these reactions has values of 10-12 and 19-21 kcal per mol at temperature ranges above and below the break temperature, respectively. The break, however, was not observed if 30% glycerol was added to the microsomes. The Arrhenius plot of the microsomal NADPH-cytochrome c reductase activity also did not show any break. The implications of these observations in relationship to the fluidity of the membrane, the translational mobility of membrane enzymes, and the rate of monoxygenase reactions are discussed. PMID- 16250347 TI - Glucose transport inhibition by proteolytic degradation of the human erythrocyte membrane inner surface. AB - Treatment of the exterior surface of human erythrocytes with the proteolytic enzymes, trypsin or alpha-chymotrypsin (at 1 mg/ml), has no discernible effect on the carrier-mediated movement of glucose. However, the incorporation of either enzyme at much lower levels inside the erythrocyte by the method of reversible hemolysis leads to a progressive inhibition of the rate of glucose movement. Total inhibition eventually results at all tested concentrations of incorporated enzyme. These results strongly suggest that a protein susceptible to attack at the interior surface of the cell membrane is in some way involved in sugar transport. Polyacrylamide gel electrophoresis in sodium dodecyl sulfate showed that the spectrin band (known to be located at the inner membrane surface) gradually disappeared during the protease treatment, in close parallel with the loss in glucose transport. This was not accompanied by any appreciable modification in Band III, which has been closely identified with the glucose transport system. PMID- 16250349 TI - The role of P(i) in glycolytic inhibition of calcium ion uptake by ELD ascites tumor cells. AB - In contrast to previous investigations at 25 degrees C, glucose was shown to support 45Ca2+ uptake at 37 degrees C in intact ELD ascites tumor cells. Intact ascites tumor cells in vitro accumulated up to 5.0 micromol of 45Ca2+ per g cells dry wt. within 20 min. In the presence of 10.0 mM glucose, intracellular P(i) levels fell from 40.0 micromol x g(-1) cells dry wt. to 20.0 micromol x g(-1) cells dry wt. in 5 min. Intracellular P(i) levels were maintained by 20.0 mM extracellular Tris-P(i). 45Ca2+ uptake was inhibited in P(i)-depleted cells, even though the metabolic rate (as measured by Q(lactate)) and energy state (as measured by ATP levels) were at acceptable levels. Evidence has been presented suggesting that previous reports of glucose inhibition of calcium uptake can be attributed to a competition for available intracellular P(i) between glycolytic processes and the mitochondrial calcium uptake mechanism. PMID- 16250350 TI - On cell membrane lipid fluidity and plant lectin agglutinability. A spin label study of mouse ascites tumor cells. AB - The fluidity of the plasma membrane of Sarcoma 180 mouse ascites tumor cells has been studied in viable cells using fatty acid spin labels. The order parameter was found to vary from 0.61, approximately four carbon bond lengths removed from the membrane surface, to 0.47 approximately eleven bond lengths removed at 22 degrees C and from 0.55 to 0.33 at 37 degrees C. Thus these cells show similar membrane fluidity to that found in other mammalian cells with the exception of human erythrocytes which are less fluid. The concanavalin A mediated agglutinability of Sarcoma 180 cells was altered by the addition of cytochalasin B and the fluidity was found to be the same as in unaltered cells. PMID- 16250348 TI - Surface properties of membrane systems. Transport of staphylococcal delta-toxin from aqueous to membrane phase. AB - Hemolytic delta-toxin from Staphylococcus aureus was soluble in either water, methanol or chloroform/methanol (2 : 1, v/v). The toxin spread readily from distilled water into films with pressures (pi) of 10 dynes/cm on water and 30 dynes/cm on 6 M urea; from chloroform/methanol it produced 40 dynes/cm pressure on distilled water. The toxin adsorbed barely from water (pi = 1 dyne/ cm) but it did rapidly from 6 M urea (pi = 35 dynes/cm). The protein films had unusually high surface potentials, which increased with the film pressure and decreased with increasing both pH and urea concentration in the aqueous phase. The fluorescence of 1-aniline 8-naphthalene sulfonate with delta-toxin was much greater than that with RNAase and dipalmitoyl phosphatidylcholine itself, indicating probably a marked lipid-binding character of the toxin. By circular dichroism the alpha-helix content of delta-toxin was 42% in water, 45% in methanol, 24% in 6 M urea. Infrared spectroscopy showed predominant alpha-helix in both 2H2O and deuterated chloroform/methanol as well as in films spread from either solvent on 2H2O. In spreading from 6 M [2H]urea, in which the major infrared absorption was that of [2H]urea with peaks at 1600 and 1480 cm(-1), the delta-toxin film showed prevalently non-alpha-helix structures with major peak intensities at 1633 cm(-1) > 1680 cm(-1), indicating the appearance of new beta aggregated and beta-antiparallel pleated sheet structures in the film. The data prove that (1) high pressure protein films can consist of alpha-helix as well as non-alpha-helix structures and, differently from another cytolytic protein, melittin, delta-toxin does not resume the alpha-helix conformation in going into the film phase from the extended chain in 6 M urea; (2) conformational changes are important in the transport of proteins from aqueous to lipid or membrane phase; (3) delta-toxin is by far more versatile in structural dynamics and more surface active than alpha-toxin. PMID- 16250351 TI - Cardiac sarcolemma of the hamster. Enrichment of the (Na+ + K+)-ATPase. AB - Cardiac sarcolemma was prepared from normal hamsters using gentle homogenization, extraction with 0.6 M KCl and continuous density gradient centrifugation. The final fraction exhibited high (Na+ + K+)-ATPase activity (24 micromol/mg per h) and contained minimal contamination from mitochondria, myofibrils and lysosomes. PMID- 16250352 TI - Direction of proton translocation in proteoliposomes formed from purple membrane and acidic lipids depends on the pH during reconstitution. AB - The reconstitution of proton pumping activity in proteoliposomes formed by brief sonication of purple membrane and lipid dispersions was studied as a function of pH. Proteoliposomes reconstituted using cardiolipin showed light-dependent proton extrusion when formed at a pH below 2.75 and proton uptake when formed above pH 2.75. Several other acidic lipids including halobacterial lipids behaved similarly. The experiments suggest that the degree of dissociation of the lipid phosphate groups determines the preferential orientation of bacteriorhodopsin in reconstituted proteoliposomes. PMID- 16250353 TI - Ion tracer flows and flux ratios in heterogeneous membranes. AB - Ion tracer flows and flux ratios at various electrical potentials were studied in heterogeneous membranes comprising parallel pathways of different intrinsic resistance. The total resistance to net flow exceeded the tracer exchange resistance, and the flux ratio was abnormal, as in exchange diffusion commonly attributed to a mobile carrier. PMID- 16250355 TI - Shattered lives. PMID- 16250354 TI - Free carboxylate groups required for transport of neutral amino acids by the Ehrlich ascites-tumor cell. AB - Although uncharged structures analogous to the carboxyl group of the amino acid molecule have served in place of that group for transport into some cells, we find that substitution either by the carboxamide group or by the chloromethyl ketone group eliminates inhibition of transport Systems A or L of the Ehrlich ascites tumor cell. Comparison of the loss of System L transport on acidification shows that the pH at which this loss occurs is correlated with pK'1 of the amino acid substrate, suggesting that transport is terminated by protonation of the site-bound carboxylate group. PMID- 16250356 TI - Liability crisis: is profiteering the problem? PMID- 16250357 TI - When patients won't speak up. PMID- 16250358 TI - Taking your new medical degree in a new direction. PMID- 16250359 TI - Is Utah the new Oregon? PMID- 16250360 TI - If a patient wants to tape the visit. PMID- 16250361 TI - Sued? Calm down! Mouthing off about it can get you into more trouble than you're already in. PMID- 16250363 TI - Don't fill out disability forms. PMID- 16250362 TI - Retirement plans are lagging. PMID- 16250364 TI - ECG of the month. ECG in a man with a fracture of the femoral neck. Left ventricular enlargement and hyperkalemia. PMID- 16250365 TI - Radiology case of the month. Unusual inflammatory etiology of a lesion in the distal femur. Blastomycosis osteomyelitis. PMID- 16250366 TI - Pathology case of the month. Tumor of the pancreas in a young woman. Solid pseudopapillary tumor (SPT) of the pancreas. PMID- 16250367 TI - A middle-aged woman with an anterior mediastinal mass and respiratory failure. AB - The discovery of an anterior mediastinal mass presents a challenge to the physician. In addition to distinguishing between benign and malignant conditions, it is important to recognize the potential for associated paraneoplastic phenomena. We present a case of a malignant thymoma associated with myasthenia gravis. PMID- 16250368 TI - Initial treatment patterns for clinically localized prostate cancer and factors associated with the treatment in Louisiana. AB - This study investigated treatment patterns among Louisiana residents diagnosed with clinically localized prostate cancer in 2001 and factors that may be associated with the treatment. The differences in the initial treatment between 1997 and 2001 were also examined. The data were collected from hospital medical records, supplemented by information from freestanding radiation centers and physicians' offices. We assessed the associations of initial treatment with demographic factors such as age, race, health insurance status, type of healthcare facility, area of residence, county poverty, and clinical factors such as Gleason score, PSA, and comorbidity in univariate and logistic multivariate regression analyses. Our study found that patients clinically diagnosed with localized prostate cancer in 2001 received the following treatments: radical prostatectomy (41.4%), radiation (29.7%), hormone (16.2%), or watchful waiting (11.9%). White patients, older patients, patients with private insurance and patients diagnosed or treated in hospitals were more likely (p < 0.05) to receive aggressive therapy (i.e., radical prostatectomy or radiation) than others after controlling for the demographic and clinical factors. Poverty level and comorbidity were inversely associated with receiving aggressive therapy in univariate analysis. But after adjusting for other factors, these associations were no longer statistically significant. Patients with elevated PSA and high Gleason scores were less likely to receive radical prostatectomy even after the adjustment. From 1997 to 2001, utilization of radiation and hormonal therapies increased, and watchful waiting decreased among newly diagnosed prostate cancer patients. Utilization of radical prostatectomy showed no significant change over time. PMID- 16250369 TI - Correlation of different 5-FU regimens and time to surgery with pathological complete response in locally advanced rectal cancer patients treated by pre operative chemo-radiation. AB - The best pre-operative chemo-radiation regimen has still to be defined for patients with rectal cancer. We retrospectively performed a study comprised of 26 patients with T3/4 adenocarcinoma of the rectum. Eleven patients received concurrent protracted venous 5-FU infusion (PVI), 11 had bolus, 1 had 4-day infusions, and 3 did not receive concurrent chemotherapy. All patients underwent surgical resection. Different variables were analyzed in relation to pathologic complete response (pCR) and survival. The use of PVI is associated with a higher pCR rate (45.5% vs. 6.7%, p = 0.054) and a trend towards less proportion of death (p =0.091), but no difference in overall survival (p = 0.439). Patients in the PVI group received a higher total 5-FU dose during radiation (p < 0.001) and had a longer delay between radiation and surgery (p = 0.002). Further study with a larger patient population is needed to confirm our observation. PMID- 16250370 TI - Influenza in Louisiana: an analysis of Louisiana Influenza Sentinel Surveillance Data, 1998-2004. AB - Analysis of the influenza sentinel surveillance data from the Louisiana Office of Public health showed that the influenza season for Louisiana starts between October and December and ends between December and March. There is no consistent trend as to when the peak of the season occurs. The start of Louisiana's seasons ranged from one week (2002-2003 season) to two months (2001-2002) before the national start date. The finding that Louisiana's flu season starts about the same time as the national flu season is not consistent with the conventional thought that the flu season starts later in the South compared to the rest of the nation. PMID- 16250371 TI - Retained palm frond injuries: two cases of delayed diagnosis. PMID- 16250372 TI - Newborn "heel stick" screening: are you ready for the new ones? PMID- 16250380 TI - [Ethics, real or rubbish?]. PMID- 16250373 TI - Centennial year of yellow fever eradication in New Orleans and the United States, 1905-2005. AB - This article briefly details the history of the 1905 yellow fever epidemic in New Orleans and the successful and permanent eradication of The Pestilence. First the authors will give some medical and historical background regarding Bronze Jack from its transmission from West Africa by the Spanish slave trade to the New World, through the Caribbean to New Orleans, the great port city which became the Necropolis of the South. We will then focus on the summer of 1905 when New Orleans experienced what proved to be the last epidemic in the history of New Orleans and the United States, and the methods employed to combat it. This year, 2005, marks the centennial of one of the truly remarkable and critical accomplishments of medicine and public health. PMID- 16250381 TI - [Steps for an ethical decision]. PMID- 16250382 TI - [And if team rhymed with ethics?]. PMID- 16250383 TI - [Specific missions for the mobile palliative care support teams]. PMID- 16250384 TI - [Life in a palliative care unit: daily ethical demands]. PMID- 16250385 TI - [Ethics and the end of life, what help for the family?]. PMID- 16250387 TI - [Patient agent, practical guide]. PMID- 16250386 TI - [Ethical approach to Alzheimer patients at the end of life]. PMID- 16250388 TI - [Hip protectors, an efficacious way to prevent femoral neck fractures]. PMID- 16250389 TI - [Evaluation of a patient/caregiver liaison chart at home]. PMID- 16250390 TI - [Interdisciplinary cooperation, a necessity in geriatrics]. PMID- 16250391 TI - Comparison of digitized and direct viewed (analog) radiographic images for detection of pulmonary nodules. AB - The purpose of this study was to compare direct (analog) viewing of thoracic radiographs with digitized images obtained with a radiographic scanner and seven digital cameras for detection of pulmonary nodules. Direct viewing of the analog radiographs was significantly better than all digitized methods (P<0.01). Significant variations exist between the radiographic scanner and the digital cameras. The scanner (Kodak LS75) was significantly better than the Kodak DC 4800 and the Sony DSC-707 cameras (P<0.05). The Nikon 995, Canon EOS-D30, and Ricoh i500 were significantly better than the Sony DSC-707 (P<0.05). There was no significant difference between the Kodak DC 3800, Kodak DC 4800, and the Sony DSC 707. For pairwise comparison of raters (when evaluating the radiographs individually compared with the consensus), the raters matched the consensus rating from 85% to 92% with no significant difference between raters. PMID- 16250392 TI - Magnetic resonance imaging features of cervical spinal cord meningiomas. AB - The records of four dogs with cervical spinal cord meningiomas were retrospectively reviewed. Signalment, history, laboratory findings, neurological examination, and histopathological findings were evaluated. Magnetic resonance imaging (MRI) was performed using a 1.0-T superconducting magnet and T2-weighted (W) and noncontrast and postcontrast T1-W spin echo pulse sequences. Meningiomas were located at the level of the second, third, and fifth cervical vertebrae and the C2-3 intervertebral space. All meningiomas appeared as focal masses that were hyperintense to the spinal cord on T2-W images and iso- to hypointense on the T1 W images. They could be identified as intradural and extramedullary in origin based on a broad-based dural margin seen on at least one of the imaging planes and a gradual expansion of the subarachnoid space cranial and caudal to the mass, best noted on the transverse and dorsal plane images. On dorsal plane T2-W images in three dogs, expansion of the subarachnoid space adjacent to the mass appeared similar to the myelographic "golf tee" sign. All meningiomas exhibited moderate, well-defined contrast enhancement with dural tails seen in three of the four dogs. One dog had extension into the intervertebral foramen along the nerve and ipsilateral atrophy of the muscles of the neck. By differentiating the meningiomas from intramedullary tumors and by clearly depicting the extent of the masses, MRI provided valuable information about treatment options and prognosis. PMID- 16250393 TI - Anatomical study of cranial nerve emergence and skull foramina in the dog using magnetic resonance imaging and computed tomography. AB - Twenty-two magnetic resonance imaging (MRI) brain studies of different breeds of dogs were reviewed to assess the anatomy of cranial nerve (CN) origins and associated skull foramina. These included five anatomic studies of normal brains using 2-mm-thick slices and 17 studies using conventional clinical protocols with 3- or 4-mm slices on both normal and abnormal brains. Images were obtained in transverse, sagittal, and dorsal planes to allow a thorough comparison between studies. CNs II, III, V (and its divisions), and VIII were observed consistently on conventional studies. On the thin-slice studies, the origins and proximal portions of CNN IV, VII, and the group of IX, X, and XI could be seen. The origins of CNN VI and XII were not observed with certainty. In parallel, a computed tomography study of an isolated skull was performed with a thin copper wire within each of the skull foramina to determine precisely each CN exit and to facilitate recognition of the course of CNs when exiting the skull on MRI images. PMID- 16250395 TI - Assessment of location of the celiac and cranial mesenteric arteries relative to the thoracolumbar spine using magnetic resonance imaging. AB - Exact localization of thoracolumbar lesions can be harder with magnetic resonance (MR) imaging than with radiography. The celiac and cranial mesenteric arteries are easily seen on MR images and are always included in sagittal thoracolumbar studies. This study was undertaken to establish whether their location was sufficiently consistent to enable them to be used as anatomic landmarks. It was found that their location varied considerably, and there was no useful relationship to breed, gender, age, or body weight. They are therefore unreliable for use in establishing initial localization of a spinal lesion although they can be helpful when comparing multiple image planes. PMID- 16250394 TI - Utility of magnetic resonance imaging for distinguishing neoplastic from non neoplastic brain lesions in dogs and cats. AB - The aim of this study was to identify magnetic resonance (MR) signs that aid differentiation of neoplastic vs. non-neoplastic brain diseases in dogs and cats. MR images of 36 dogs and 13 cats with histologic diagnosis of intracranial disease were reviewed retrospectively. Diagnoses included 30 primary and three metastatic brain tumors, 11 infectious/inflammatory lesions, three vascular, one degenerative disease, and one developmental malformation. Upon univariate analysis of 21 MR signs, there were seven that had a significant association with neoplasia: single lesion (P = 0.004), shape (P = 0.015), mass effect (P = 0.002), dural contact (P = 0.04), dural tail (P = 0.005), lesions affecting adjacent bone (P = 0.008), and contrast enhancement (P = 0.025). Increasing age was also found to be associated with neoplasia (P = 0.0001). MR signs of non-neoplastic brain diseases in dogs and cats were more variable than those of brain neoplasia. PMID- 16250396 TI - Comparison of radiographic and computed tomography lymphangiography for identification of the canine thoracic duct. AB - Standard radiographic lymphangiograms and computed tomography (CT) lymphangiograms were performed on 10 female dogs without intrathoracic disease. Positive contrast lymphagiography was performed by injection into a catheterized mesenteric lymphatic vessel, and lateral thoracic radiographs, ventrodorsal thoracic radiographs, and thoracic CTs were obtained. The number of visible ducts was recorded for each image at the midbody of the ninth thoracic vertebra (T9) through the first lumbar vertebra (L1). Data were combined for all dogs at each data acquisition point. Data were analyzed by comparing data from all three images independently, and then by combining data for the radiographs and comparing the study with the highest number of visible duct branches to the CT. Significant differences in numbers of branches were found at T11 and L1. This study suggests that CT may be able to quantify branches of the thoracic duct more accurately than standard radiographic lymphangiography. PMID- 16250397 TI - Sublumbar abscess and diskospondylitis in a cat. AB - Diskospondylitis is uncommon in cats. We describe a cat with diskospondylitis of the L7-S1 intervertebral disk, and a concurrent sublumbar abscess. Radiographic, computed tomographic and ultrasonographic findings are presented. Aerobic and anaerobic cultures of blood and spinal fluid yielded no growth. Aerobic and anaerobic urine cultures resulted in growth of an Enterococcus sp. and Clostridium perfringens, respectively. The cat was successfully treated with enrofloxacin and amoxicillin/clavulanic acid. Clinical signs resolved completely, and based on follow-up ultrasonography there was no remaining evidence of the sublumbar abscess. Etiologic agents and outcome from other cats with diskospondylitis are reviewed. PMID- 16250398 TI - Vertebral heart size ranges specific for whippets. AB - To assess the influence of breed, breeding lines, and training on heart size, the vertebral heart size (VHS) was evaluated on left-to-right lateral, right-to-left lateral, dorsoventral, and ventrodorsal thoracic radiographs from 44 whippets free from cardiac and pulmonary disease. In lateral views, the VHS was 11.0 +/- 0.5 vertebrae (mean +/- SD) on right-to-left lateral and 11.3 +/- 0.5 vertebrae on left-to-right lateral radiographs, being larger than the 9.7 +/- 0.5 vertebrae proposed by Buchanan (P<0.0001). The VHS on left-to-right lateral views was larger than on right-to-left lateral views (P<0.0001). The VHS was 10.5 +/- 0.6 vertebrae on dorsoventral radiographs and 11.1 +/- 0.6 vertebrae on ventrodorsal radiographs. Both values were larger than the 10.2 +/- 1.5 vertebrae (dorsoventral) (P<0.0082) or 10.2 +/- 0.8 vertebrae (ventrodorsal) (P<0.0001) proposed by Buchanan. In addition, the VHS on ventrodorsal views was larger than on dorsoventral views (P<0.0001). Dogs out of racing pedigree lines had a significantly larger VHS than those out of show pedigree lines, and trained dogs had a significantly larger VHS than nontrained dogs. Because most trained dogs came out of racing pedigree lines, and all nontrained dogs came out of show pedigree lines, however, it is difficult to determine whether the higher VHS for trained dogs is due to genetic influences or training, or both. In conclusion, it is important to take into account the breed and the radiographic view when evaluating heart size in thoracic radiographs of whippets to avoid overinterpretation of cardiac enlargement in these dogs. PMID- 16250399 TI - Characterization of normal and abnormal canine superficial lymph nodes using gray scale B-mode, color flow mapping, power, and spectral Doppler ultrasonography: a multivariate study. AB - The objective of this study was to characterize the ultrasonographic patterns of normal superficial lymph nodes and to evaluate whether ultrasonography can help discriminate between different lymphadenopathies (reactive, lymphoma, and metastases) in dogs. Three hundred and eighteen superficial lymph nodes in 142 dogs were studied by B-mode, color flow mapping, power, and spectral Doppler ultrasonography. Size, echogenicity, nodal border definition, presence of a nodal hilus, acoustic enhancement and distribution of vascular flow, as well as perfusion indices were measured. Multivariate statistics using discriminant analysis was used to determine which parameters can be used to predict the diagnosis of the lymph node. The size of the lymph node, distribution of vascular flow within the lymph node, and pulsatility index (PI) in combination gave a classification error of 23% for the four groups of lymph nodes. This was improved to 11% if the nodes were divided into two groups: benign and malignant. There was a significant difference in resistive index (RI) and PI between benign and malignant nodes. Cut-off values were determined using receiver operator curves, 0.68 RI and 1.49 PI. PMID- 16250401 TI - Ultrasonographic features of peritoneal cestodiasis caused by Mesocestoides sp. in a dog and in a cat. AB - Peritoneal infections caused by Mesocestoides spp. are rare in dogs and cats. Little data exist on the role of abdominal ultrasonography for diagnosis and therapy management of the disease. We describe the ultrasonographic features of peritoneal cestodiasis in a dog and in a cat. In the dog, abdominal ultrasound allowed both a presumptive diagnosis and the collection of tissue samples to confirm peritoneal larval infection. Ultrasound was also very useful for therapy management. In the second patient the ultrasonographic features of tetrathyridial infection in a cat in which the parasite was observed as an incidental finding during ovariohysterectomy are described. PMID- 16250400 TI - Characterization of lymphomatous lymph nodes in dogs using contrast harmonic and Power Doppler ultrasound. AB - Doppler ultrasound has been used in humans to determine angioarchitecture of lymph nodes as the criterion for the determination of malignancy. We hypothesized that the vascular and perfusion patterns of a canine malignant lymph node could be characterized with intravenous microbubble ultrasound contrast media and that contrast harmonic ultrasound could provide better conspicuity of the angioarchitecture when compared with Power Doppler ultrasound. In this study, 11 peripheral lymph nodes in dogs with histologically verified malignant lymphoma were imaged with fundamental ultrasound, Power Doppler ultrasound, and three contrast harmonic pulse sequences to characterize the vascular pattern and perfusion. Vascular imaging was greatly enhanced in these nodes with 2.13 times more vessels seen with contrast harmonic ultrasound compared with Power Doppler ultrasound (P < 0.01). The angioarchitecture of lymphomatous lymph nodes of dogs in this study were similar to those previously described in malignant superficial lymph nodes in human patients; 45.5% of the nodes had displacement of the central hilar vessel, 45.5% had aberrant vessels, 63.6% had pericapsular vessels, 36.4% had subcapsular vessels, and 81.8% had loss of the central hyperechoic band in fundamental sonography. Poor perfusion, indicated by a lower mean pixel intensity increase between pre- and postcontrast administration images, was seen in 36.4% of the lymph nodes while 63.6% had fair to good perfusion. The perfusion patterns in nine of the 11 lymph nodes were homogenous and two showed focal hypoperfused regions. We conclude that Power Doppler and contrast harmonic ultrasound are beneficial in accurately depicting angioarchitechture and can provide additional information in determining the presence of malignant vascular characteristics within lymphomatous nodes in dogs. PMID- 16250402 TI - Ultrasonographic features of grass awn migration in the dog. AB - The aim of this study was to describe the ultrasonographic features of grass awns in soft tissue. A 10 MHz linear transducer was used. Ultrasound images from 25 dogs (27 awns) were collected and compared with the results from water bath studies using wild oat seeds (Avena spp.) collected in the field. Wild oat seeds were the most common grass awn found in soft tissue of dogs. Ultrasonographically grass awns appeared as a double/triple spindle-shaped echogenic interface within soft tissue. The same appearance was observed in water bath studies. In four dogs, the grass awn was removed surgically with a clamp introduced into a fistulous tract, using sonographic guidance. The grass awn was not found surgically in only three dogs, suggesting more attention during surgery. Ultrasonography is a useful diagnostic imaging technique to identify grass awns within soft tissue. PMID- 16250403 TI - Influence of normovolemic anemia on Doppler-derived blood velocity ratios of abdominal splanchnic vessels in clinically normal dogs. AB - Doppler spectra of the abdominal aorta (AAo), cranial mesenteric artery (CMA), celiac artery (CA), and left renal artery (LRA) were obtained from 11 fasted, clinically healthy, conscious Beagles before and after inducing severe acute normovolemic anemia (mean +/- standard deviation hematocrit 16.0 +/- 0.77%). Peak systolic, end diastolic, and time-averaged mean velocities were measured. The different vessels were compared with each other. Peak systolic velocity ratio and time-averaged mean velocity ratio of splanchnic vessels to corresponding variables of the AAo were computed and compared between and within vessels during physiologic and anemic states. There was no difference between LRA and AAo, CMA or CA regarding time-averaged mean velocity, time-averaged mean velocity ratio, or end diastolic velocity during the physiological state. During the anemic state, LRA mean time-averaged mean velocity (P < or = 0.008) and mean end diastolic velocity (P < or = 0.041) were significantly lower than those of AAo, CMA, and CA. Mean time-averaged mean velocity ratio of the LRA was also significantly (P < or = 0.004) lower than the CMA and CA ratios, and significantly (P = 0.014) lower during anemic state than physiologic state of the same vessel. End diastolic and time-averaged mean velocities of the AAo, CMA, and CA increased proportionally during anemia, but there was a relatively less increase in the same variables of the LRA, suggesting less increase in blood flow. Doppler-ratios allowed a noninvasive comparison between splanchnic and aortic hemodynamics. Velocity ratios might be useful for clinical detection of relative hemodynamic changes between different vessels. PMID- 16250404 TI - Comparison of glomerular number and specimen length obtained from 100 dogs via percutaneous echo-assisted renal biopsy using two different needles. AB - Our objective was to evaluate possible differences in the number of glomeruli and length of renal biopsies collected in canine subjects by two different types of biopsy needles: a semiautomatic 18-gauge Trucut and an automated 18-gauge Jamshidi modified (Biopince). One hundred biopsy samples obtained from dogs of different ages and gender affected by different nephropathies were evaluated retrospectively. All animals were biopsied using one of the two different needles. Biopsies were performed under ultrasound guidance and evaluated by a single pathologist. Statistical analysis was performed to evaluate possible differences in the number of glomeruli and length of renal biopsies collected comparisons were determined between subgroups of dogs with or without the identification of renal interstitial infiltrates and/or fibrosis. Neither the mean difference of the number of glomeruli nor the length of tissue sample collected with the different needles was significantly different. Likewise, the average biopsy length did not differ in dogs with or without renal interstitial infiltrate in animals biopsied with either biopsy needle. Both the Biopince and the Trucut devices provide diagnostically adequate biopsy renal specimens using ultrasound-guidance. PMID- 16250406 TI - Average cost per person victimized by an intimate partner of the opposite gender: a comparison of men and women. AB - Differences in prevalence, injury, and utilization of services between female and male victims of intimate partner violence (IPV) have been noted. However, there are no studies indicating approximate costs of men's IPV victimization. This study explored gender differences in service utilization for physical IPV injuries and average cost per person victimized by an intimate partner of the opposite gender. Significantly more women than men reported physical IPV victimization and related injuries. A greater proportion of women than men reported seeking mental health services and reported more visits on average in response to physical IPV victimization. Women were more likely than men to report using emergency department, inpatient hospital, and physician services, and were more likely than men to take time off from work and from childcare or household duties because of their injuries. The total average per person cost for women experiencing at least one physical IPV victimization was more than twice the average per person cost for men. PMID- 16250405 TI - Effects of acepromazine on three-phase 99mTc-MDP bone imaging in 11 horses. AB - Horses undergoing skeletal scintigraphy can have decreased radiopharmaceutical bone uptake in the limbs. This reduces the diagnostic value of the scan. The aim of the present study was to measure the changes in count density caused by vasodilatation and increased blood flow associated with intravenous injection of acepromazine during bone scintigraphy in normal horses. A three-phase bone scan was performed twice in 11 adult horses to study the effects of acepromazine on the count density of the resultant scintigrams. With acepromazine, there was a statistically significant mean difference of 12 s for initial blood flow and 21 s for peak flow. The time to initial blood flow and time to peak flow occurred earlier for the scans in which acepromazine was used. There were no significant differences in the bone to soft tissue ratios during the soft tissue and bone phases of the scan between procedures. Intravenous administration of acepromazine increases peripheral blood flow causing an earlier onset of the vascular phase during the three-phase bone scan. Acepromazine did not increase the count density of the bone phase scintigrams. As expected, the vasodilatation and increased blood flow associated with intravenous injection of acepromazine affected the count density of the vascular phase of the bone scan. PMID- 16250407 TI - Unidirectional and bidirectional intimate partner violence among White, Black, and Hispanic couples in the United States. AB - This article describes the rates and risk factors of unidirectional and bidirectional intimate partner violence (IPV) among White, Black and Hispanic couples in the US. Subjects constitute a multistage area probability sample representative of married and cohabiting couples from the 48 contiguous United States. Results indicate that most couples reporting violence engage in bidirectional IPV. Blacks are more likely than Whites to report bidirectional IPV. In addition, severe unidirectional and bidirectional IPV are more common among Blacks and Hispanics. The results also indicate that predictors of IPV vary depending on whether it is unidirectional or bidirectional. Unidirectional female to-male IPV and bidirectional IPV are more associated with the characteristics of the female. In contrast, the occurrence of unidirectional male-to-female IPV is associated with a single risk factor, male childhood physical abuse. These findings highlight the importance of distinguishing different types of IPV and have important epidemiological and prevention implications. PMID- 16250408 TI - Validation of the subtle and overt psychological abuse scale: an examination of construct validity. AB - The Subtle and Overt Scale of Psychological Abuse (Marshall, 1999a) is a measure designed to examine previously unevaluated forms of psychological abuse. The scale was originally divided into seven subscales (overt: dominance, indifference, monitoring and discrediting; subtle: undermining, discounting, isolating). A sample of 172 women was used to evaluate the construct validity of this measure. In other words, the internal structure of the measure, its relation to other measures of physical and psychological abuse, and its relationship to other psychological outcomes. Empirical and theoretical approaches were used to replicate and examine the dimensions of this scale. Results revealed that this scale represents a unidimensional construct that is highly correlated to other measures of psychological abuse. Implications for future research and practice are discussed. PMID- 16250409 TI - Correlates of women's sexual assault disclosure to informal and formal support sources. AB - Deciding which people to tell about sexual assault is an important and potentially consequential decision for sexual assault survivors. Women typically receive many different positive and negative reactions when they disclose sexual assault to social support sources. A diverse sample of adult sexual assault survivors in the Chicago area was surveyed about sexual assault experiences, social reactions received when disclosing assault to others, attributions of blame, coping strategies, and PTSD. Analyses were run to identify demographic, assault, and postassault factors differentiating women disclosing to informal support sources only from those disclosing to both informal and formal support sources. Women disclosing to both formal and informal support sources experienced more stereotypical assaults, had more PTSD symptoms, engaged in less behavioral self-blame, and received more negative social reactions than those disclosing to informal support sources only. Implications for future research and intervention are discussed. PMID- 16250410 TI - The dynamic nature of the drug use/serious violence relationship: a multi-causal approach. AB - Relying on historical research, a longitudinal data set, and multivariate analyses, the drug-violence relationship is scrutinized. A proposed model is tested and supported, indicating that attitudes toward violence, gender, neighborhood problems, minor delinquency, and victimization were persistent correlates that must be considered within the drug-violence relationship. Parental attachment and importance, exposure to delinquent peers, and drug dealing were also important. Both licit and illicit drug use were significant within the models, although the relationship changed from year to year. In year 1, youth who used drugs reported more violence. In year 2, youth who were not using drugs reported more violence. Association with delinquent peers and initial involvement in drug dealing were likely explanations for this transition. Findings offer support for prevention efforts that disrupt drug markets and target male youth who are involved in crime and drugs, repeatedly victimized, associating with delinquent peers, and developing attitudes favorable toward the use of violence. PMID- 16250411 TI - Angry drivers: a test of state-trait theory. AB - Tested hypotheses from state-trait theory applied to anger while driving. College student drivers high in trait driving anger were compared to drivers low in trait driving anger. High anger drivers were more frequently angered in day-to-day driving (frequency hypothesis). They reported more intense anger in their most angering driving situations, when visualizing provocative driving events, and in day-to-day driving (intensity hypothesis). Driving diaries and surveys showed they engaged in more aggressive behavior and expressed their anger through more verbal, physical, and vehicular means (aggression hypothesis). They reported handling of their anger less well when visualizing provocative events and on the Adaptive/Constructive Expression scale (reduced adaptive expression hypothesis). They engaged in risky behavior (risky behavior hypothesis) and experienced more moving violations, close calls, and losses of concentration, but not more major or minor accidents (partial support for crash-related outcomes hypothesis). High anger drivers were more generally angry and impulsive and employed more negative, less controlled forms of general anger expression. Results supported state-trait theory and added to the literature showing that high anger drivers have some other psychological and behavioral characteristics that may interact negatively with anger behind the wheel. PMID- 16250413 TI - Watch out for our children. They could be your own. PMID- 16250412 TI - September 11 attacks: prior interpersonal trauma, dysfunctional cognitions, and trauma response in a midwestern university sample. AB - Undergraduate college students (N = 133) were assessed for posttrauma and depressive symptomatology, posttrauma cognitions, and previous traumatic experiences 1 month after the September 11 attacks in the United States. Negative beliefs regarding oneself and the world mediated the relationship between prior interpersonal trauma and acute trauma symptoms. Acute depression acted as a mediator between prior interpersonal trauma and negative cognitions of oneself and self-blame cognitions. The results suggest that even in a nonclinical, nontreatment seeking college sample, prior traumatic events can play an important role in an individual's short-term adjustment following indirect exposure to a significant trauma. The differential relationship of negative beliefs on acute trauma symptoms and depression is discussed. PMID- 16250415 TI - Promotion of disease and corrosion of medicine. PMID- 16250414 TI - Childhood sexual abuse and family physicians. PMID- 16250416 TI - Praise for family physicians. PMID- 16250417 TI - At his age. Once upon a time. . . PMID- 16250418 TI - Prolonged exposure to angiotensin-converting enzyme inhibitors during pregnancy. Fetal toxicity could be reversible. AB - QUESTION: I read in a Motherisk Update that angiotensin-converting enzyme (ACE) inhibitors are contraindicated during pregnancy. Many women, however, do not know they are pregnant for quite some time after conception. One of my patients was taking ACE inhibitors for 3 to 4 months while she was pregnant. How should I advise her? ANSWER: The deleterious effects ACE inhibitors have on fetuses were seen only after exposure during the second and third trimesters and were mostly secondary to renal damage. These effects can be reversed, as described in this Motherisk Update. PMID- 16250419 TI - Female athlete triad. Each of the following statements about treatment for menstrual disturbances and decreased bone mineral density in female athletes is true except. PMID- 16250420 TI - Screening mnemonic for generalized anxiety disorder. PMID- 16250421 TI - Approach to evaluation of sexual assault in children. Experience of a secondary level regional pediatric sexual assault clinic. AB - OBJECTIVE: To determine whether a 3-year-old girl, brought to an after-hours clinic because her mother was concerned, had been assaulted by her father during a weekend visit. SOURCES OF INFORMATION MEDLINE: was searched using the key words child, sexual assault/abuse, and expectations. Recent textbooks on childhood sexual assault and abuse were consulted. The secondary-level regional pediatric sexual assault clinic's experience over 1 year was reviewed. Articles in the literature generally provide level II evidence. MAIN MESSAGE: The literature review and the clinic's experience both indicated that specialty centres for child sexual assault and abuse rarely produce positive physical findings that conclusively confirm or rule out sexual assault, especially when children are asymptomatic and not in an acute state. Primary care practitioners can use a brief history and physical examination to decide on the next level of care and determine the urgency of referral. Urgent assessment of children thought to have been abused or assaulted is required when children disclose assault (especially with genital-genital contact or ejaculation); when children have acute pain, bleeding, or discharge; when results of a physician's examination are abnormal; or when parents are extremely distressed. CONCLUSION: Family physicians have a pivotal role in evaluation of childhood sexual assault or abuse. Knowledge of the outcomes of evaluation is crucial to understanding when and how to refer. PMID- 16250423 TI - Treating children's mental health problems. Collaborative solutions for family physicians. PMID- 16250424 TI - To Ireland and back. PMID- 16250422 TI - Approach to infected skin ulcers. AB - OBJECTIVE: To review the diagnosis and management of infected chronic skin ulcers. SOURCES OF INFORMATION: Cochrane database, MEDLINE, and Google were searched for clinical practice guidelines (CPGs) for wound care. Most recommendations found in the CPGs had level II or III evidence. Expert and consensus opinion from the Canadian Chronic Wound Advisory Board and the International Wound Bed Preparation Advisory Board were also used. MAIN MESSAGE: Bacteria in skin ulcers act along a continuum from contamination through colonization and critical colonization to infection. Critical colonization is not always associated with overt signs of infection but can result in failure to heal, poor-quality granulation tissue, increased wound friability, and increased drainage. Good-quality swab samples should be an adjunct to clinical acumen, not a primary strategy for diagnosis. Iodine and silver-based dressings, topical antibiotics, and systemic antibiotics can be helpful. CONCLUSION: Diagnosis of chronic wound infection is based on clinical signs and a holistic approach to patients. More research into assessment and treatment of skin ulcer infection is needed. PMID- 16250425 TI - Choice: fad or fashion? PMID- 16250426 TI - Midwife shortages 'endanger lives'. PMID- 16250427 TI - Thirty thousand pregnant women sacked every year. PMID- 16250428 TI - Coagulation disorders (1). Deep vein thrombosis: breaking the silence. PMID- 16250429 TI - It's good to talk: pre- and post-birth interaction. AB - This article describes the development of evidence-based pictorial information and activity cards that can be used with parents in the antenatal and postnatal period. The focus of this project is on developing early pre- and post-birth interaction between the baby and its family. AIM: To develop a way of working with parents-to-be and their families in pregnancy and the early postnatal days that will improve interaction and communication to maximise the baby' full potential. OBJECTIVES: To develop information cards and activities that are evidence based to support positive early interaction and communication. To support parent baby attachment prior to birth. To support parent-baby attachment post birth. To establish good patterns of parent-baby communication from an early age. To provide the baby with maximum stimulation to aid brain development, physical development and growth, hearing and communication skills (Verney 1981). To increase parents' self-esteem. To involve family members. To reduce sibling rivalry. PMID- 16250430 TI - Asking the question: antenatal domestic violence. PMID- 16250431 TI - Fears and feelings in second-time pregnancy. PMID- 16250432 TI - Show some respect! Young mothers and midwives. PMID- 16250433 TI - Discontinuing epidural analgesia in second stage. PMID- 16250434 TI - Midwifery basics: postnatal care. Postnatal care in the community. PMID- 16250435 TI - The trusty toilet. PMID- 16250436 TI - Good vibrations. PMID- 16250437 TI - [Quantitative structure-retention relationships of monosubstituted alkanes by dividing its molecular structure into substructure]. AB - In order to investigate the quantitative structure-retention relationship in gas chromatography (GC) , the molecular structure of monosubstituted alkane RX (X = halogen, OH, SH, NH2) is divided into two parts, R and X, to obtain molecular structure parameters, and the retention times in GC for 37 monosubstituted alkanes RX were determined. It was proposed that the retention time in GC is affected by three main factors for RX compounds, alkyl group R, substituted group X, and interaction between R and X. Using four parameters, the eigenvalue of bonding orbital-connection matrix EVM, the polarizability effect index of alkyl group PEI, the mass content for substituted group X, and the partial charge deltaN(H) on hydrogen atom of the group X, a quantitative structure-retention correlation model with correlation coefficient (r) of 0.9948 and standard deviation (S) of 0.0991 was obtained for the 37 RX compounds. The model obtained has good predictive and extrapolation ability. The predicted retention indexes are in good agreement with the experimental ones for alcohols. PMID- 16250438 TI - [Quantitative relationship between gas chromatographic retention indices and structural parameters of polychlorinated naphthalenes]. AB - The structural and thermodynamic properties of 76 polychlorinated naphthalenes (PCNs) were fully computed at B3LYP/6-31G* level. Both structural and thermodynamic parameters of PCNs obtained were consequently taken as theoretical descriptors and correlated with their gas chromatographic retention indices (RI), so as to develop the relevant quantitative structure-retention relationship (QSRR) regression model (model I) with r2 of 0.9957, which possesses high correlation, high predictive power and clear physical interpretations. Secondly, another linear QSRR model (model II) was achieved by employing the number and position of chlorine substitution as descriptors, of which r2 was 0.9967, and also the main factors affecting the retention time of PCNs were investigated. PMID- 16250439 TI - Relationships of surfactant head group weight fraction and some polarity terms by gas chromatography. AB - Polarity, partition coefficient (K) , methanol carbon number of surfactant ((C(MeoH))S), and methanol carbon number of surfactant head group ((C(MeOH)) HG) are measured on six alkanolamides and five polyoxyethylenated long chain amines as stationary phases. From the measured methanol carbon numbers, polarity indices, (IP)S and (IP)HG, are calculated. The determined polarity terms are plotted against the head group weight fraction (f(HG)) of the investigated surfactants and several equations have been developed. The study reveals that the molecular structural gap between alkanolamides and polyoxyethylenated long chain amines diminished when HLB numbers of these surfactant classes are plotted against f(HG) values. Consequently, a general equation relating HLB and f(HG) is obtained. PMID- 16250440 TI - Determination of polarity terms of some nitrogen-containing surfactants and simulated hydrophobic tail models by gas chromatography. AB - Some polarity terms of two groups of nitrogen-containing surfactants (six alkanolamides and nine polyoxyethylenated long chain amines) are measured through gas chromatography. The apparent methanol carbon number (C(MeOH)) and polarity index (IP) values are determined on the investigated surfactants as stationary phases in packed columns. Similarly, C(MeOH) and IP values are determined on simulated hydrophobic tail (SHT) models. The obtained results reveal that the introduction of SHT approach permits the distinction between the polarities of different surfactants and their head groups. The measured polarity terms are discussed as related to hydrophile-lipophile balance (HLB) number and the hydrophobic group carbon number (RCN). Some equations relating the measured polarity values and these variable have been developed. PMID- 16250441 TI - [Isolation and identification of glycinol from Glycine max [L.] Merri]. AB - As one of the main phytoalexins and phytoestrogens, glyceollin is an important prenylflavonoid in Glycine max [L.] Merri. (soybean). Many kinds of elicitors can be used to induce its accumulation. Its biosynthesis pathway is commonly used to study the characteristics of prenyltransferase, which catalyzes the prenylated reaction happening in a very few plant families in nature. Glycinol, the direct precursor of glyceollin, is necessary to study the prenylated reaction in soybean. In comparing with the other elicitors to elicit the glycinol accumulation in soybean cotyledons, AgNO3 is the most effective elicitor. Exposure of 6-8 days old cotyledons to 0.01 mol/L AgNO3 and incubation for 24 h result in the accumulation of 256 microg (glycinol)/g (fresh weight). The glycinol was extracted by methanol. Then the isolation and purification were conducted by preparative high performance liquid chromatography. Instead of 100% acetonitrile-0.1% formic acid as the elution system, the extract was eluted by 100% methanol-0.1% formic acid. Glycinol eluted earlier than daidzin under this system and decreased the disturbance from the large amount of daidzin. Identification was performed by comparing the mass spectrum (liquid chromatography/quadrupole-time of flight) and ultraviolet spectrum with those of the standard. At last, 100 mg purified glycinol was obtained from 390 g of fresh material. PMID- 16250442 TI - [Determination of sildenafil and vardenafil in human plasma by high performance liquid chromatography coupled with liquid-liquid-liquid microextraction]. AB - High performance liquid chromatography coupled with liquid-liquid-liquid microextraction was developed for the simultaneous determination of sildenafil and vardenafil in human plasma. The effects of extraction solvent, the volume of organic solvent, dropsize of acceptor phase, stirring rate and extraction time on the enrichment factors of analytes were investigated. The optimized experimental conditions, 300 microL toluene as the organic phase, 2 microL 0.2 mol/L HCl as the acceptor phase, 600 r/min of the stirring rate, and 40 min of the extraction time, were gotten. Under these conditions, high enrichment factors were obtained. The linear range of studied analytes was from 5 microg/L to 1.0 mg/L. The relative standard deviation was lower than 5%. The limits of detection were 1 microg/L for sildenafil and 0.5 microg/L for vardenafil at signal-to-noise ratio of 3. The method with little solvent consumption may provide high analyte preconcentration and excellent sample clean-up, and it is a sensitive and suitable method for simultaneous determination of the above two substances in human plasma. PMID- 16250443 TI - [Analysis of nonylphenol, octylphenol and bisphenol A in animal tissues by liquid chromatography-tandem mass spectrometry with accelerated solvent extraction]. AB - A comprehensive analytical method based on liquid chromatography-electrospray ionization tandem mass spectrometry with negative ionization mode has been developed for measuring alkylphenols (AP) and bisphenol A (BPA) in animal tissues. Samples of animal tissues were extracted by accelerated solvent extraction with dichloromethane. Sample concentration and purification were performed using an OASIS NH2 solid extraction cartridge. The effects of mobile phase and additives on ionization were assessed. The recoveries for the compounds ranged from 88% to 101% and the relative standard deviations were below 15%. The detection limits of the method under multiple-reaction monitoring mode were 0.3, 0.05 and 0.1 microg/kg for BPA, nonylphenol (NP) and octylphenol (OP), respectively. The contents of NP ranging from 0.49 to 55.98 microg/kg were found in 27 real samples of animal tissues from Beijing market. The results indicate that the endocrine disrupting nonylphenol is ubiquitous in food of animal origin. Nonylphenol was found in all fish samples with concentration levels ranging from 9. 13 to 55.98 microg/kg. PMID- 16250444 TI - [Determination of peroxides in environmental samples by high performance liquid chromatography with fluorescence detection]. AB - A high performance liquid chromatographic (HPLC) method with fluorescence detection was developed for the determination of hydrogen peroxide and organic peroxides in environmental samples, and the method has been applied to peroxides detection in urban air and rain samples. The analytical sensitivity was improved. Post-column derivatization involved the oxidation of peroxides to a fluorescent dimer using p-hydroxyphenylacetic acid, a reaction catalyzed by hemin. The optimal excitation wavelength was 315 nm, while the emission wavelength was 400 nm. The temperature of the reaction coil was controlled at about 30 degrees C. Based on the ratio of signal to noise of 3, the detection limits were 4.0 x 10( 9) mol/L for hydroperoxide (H2O2), 4.1 x 10(-8) mol/L for methylhydroperoxide (MHP) and 6.7 x 10(-8) mol/L for ethylhydroperoxide (EHP) for aqueous samples. While the corresponding detection limits were 2.4 ng/m3 for H2O2, 35.2 ng/m3 for MHP, and 74.4 ng/m3 for EHP for air samples. Air samples were collected by Horibe cold trap at the temperature of about -90 degrees C. The results show that H2O2, hydroxymethyl hydroperoxide (HMHP), MHP, and EHP were the major peroxides in air, and that peroxyacetic acid (PAA) was occasionally detected. In rain samples, the two major peroxides were H2O2 and HMHP. PMID- 16250445 TI - [Study on trace component in sex pheromones of Dendrolimus spp]. AB - Two compounds were isolated, as sex pheromone components, from the abdominal tips of the female pine caterpillar moth, Dendrolimus kikuchii. The major component was identified as (Z,E)-5,7-dodecadien-1-yl acetate by gas chromatography and mass spectrometry. There are some difficulties to elucidate the structure of the minor component due to its trace and coelution with other components. The derivatives of alkaline methanolysis and reacetylation of pheromone gland extracts of D. kikuchii were analyzed by high-resolution gas chromatography, which was performed to verify the functional group and stereo isomers of the trace component in the pheromone gland extracts. The trace component was characterized as (Z,E) -5,7-dodecadienol via microchemical reaction. The advantages of the conversion of acetates to corresponding alcohols or of alcohols to the corresponding acetates in identifying the trace component of pheromone gland extracts of D. kikuchii were discussed. The importance of identifying the trace component in pheromone chemical communication system of insects is emphasized. PMID- 16250446 TI - [Quantitative analysis of cantide in plasma by capillary gel electrophoresis]. AB - Cantide is a 20-mer antisense phosphorothioate oligonucleotide that inhibits telomerase catalytic subunit hTERT, pharmacologic results showed that it had promising antitumor activity. In order to study the pharmacokinetic properties of Cantide, a capillary gel electrophoretic (CGE) method with internal standard was used for the determination of Cantide in rat plasma. Cantide and the internal standard had approximately equal percentage of base composition. Extraction of the phosphorothioate oligonucleotides from plasma was accomplished using two solid-phase extraction columns, a strong anion-exchange column to remove plasma proteins and lipids, followed by a reversed-phase column to remove plasma salts. A second desalting step, achieved by dialysis utilizing a membrane, was required to remove residual ionic material from the extracted sample. The size of the capillary column was 31 cm x 100 microm i.d. with an effective length of 20 cm. The running buffer was a mixture of Tris-boric acid-urea (pH 8.5). The calibration curve was linear in the range of 12.5 - 400 mg/L, with correlation coefficient (r) of 0. 999 8. Intra-day and inter-day relative standard deviations (RSDs) for the extracted samples were 0.398% - 2.46% and 2.75% - 6.07%, respectively. The range of recoveries was 99.53% - 102.1%. The results demonstrate the high accuracy, stability and reproducibility of the procedure. PMID- 16250447 TI - [Immobilized artificial membrane chromatography and its application in profiling the drug membrane transport]. AB - Immobilized artificial membrane chromatography (IAMC) is a system of covalently bonding monolayer of phospholipid analogs onto the solid surface of silica particles, thus establishing the emulating process of drug-cellular membrane interactions in terms of chromatographic method, and is applied to profile the drug membrane permeability and biological activity. IAMC phases prepared from phosphocholine analogues closely mimic the surface of the biological cell membrane, and the capacity factor of drugs can be used to predict the interaction between drug molecules and cell membranes. IAMC, compared with n-octanol/water, liposome/water and ODS chromatography, can simulate other forces besides hydrophobic interaction. IAMC is getting more and more profound research because of its convenience and high efficiency. The primary screening by means of IAMC in the early stage of drug development would effectively lower failure in the preclinical and clinical development, since efficacy and safety are related in the larger part to membrane permeability. The interaction mechanism of drug-IAM and its profiling in membrane transport are reviewed. PMID- 16250448 TI - [Studies on chromatographic properties of 2,4,6-trinitrophenol-modified zirconia magnesia stationary phase for the separation of fullerenes ]. AB - A stationary phase for the separation of C60 and C70 was prepared by modifying zirconia-magnesia composites with 2,4,6-trinitrophenol. The modified composite was characterized by using elemental analysis, diffused reflectance Fourier transform infrared spectroscopy and specific surface area. The effects of the toluene content in toluene-cyclohexane mobile phase and the column temperature on the separation of C60 and C70 were examined. Meanwhile, the separation of fullerenes including 3% high fullerenes was investigated at the temperature of 348 K using pure toluene as the mobile phase. The results showed that the stationary phase of 2,4,6-trinitrophenol-modified zirconia-magnesia composite exhibits advantage in the separation of C60 and C70, and strong temperature dependence. The retention times of C60 and C70 and their separation factor on the stationary phase increase with the increase of the column temperature. The stationary phase is a potential packing material for separation of fullerenes in preparative scale. PMID- 16250449 TI - [Analysis of ginsenosides in ginseng by liquid chromatographyatmospheric pressure chemical ionization mass spectrometry]. AB - Analysis of ginsenosides using high performance liquid chromatography-tandem atmospheric pressure chemical ionization mass spectrometry (HPLC-APCI/MS) has been carried out. HPLC analysis was performed on a reversed-phase C18 column using mobile phase of acetonitrile and water mixture under gradient elution. Effects of evaporation temperature of APCI-MS on identification of ginsenosides were investigated. APCI/MS-MS spectra of [M-H]- were further resolved to identify the structures of ginsenosides. Detection limits of ginsenosides Rb, and Rg, were determined under selected reaction monitoring (SRM) mode. Ginsenosides in Panax ginseng (white ginseng) were assayed. Results show that although APCI is a high temperature evaporative process and ginsenosides are thermolabile compounds, abundant [M-H]- ions could be detected by using APCI-MS. The intensity of [M-H]- increased with the increase of evaporation temperature, which may due to more complete evaporation of elute solution at higher temperatures or due to the ionization mechanism of APCI. The detection limits of Rb1 and Rg1 were 1.2 x 10( 13) g and 3.0 x 10(-14) g, respectively. Twenty nine ginsenosides including malonyl-ginsenosides in Panax ginseng were identified. The method developed is sensitive, reproducible and accurate. Most ginsenosides in the extracts of ginseng could be effectively identified and analyzed for their structures. PMID- 16250450 TI - [Determination of relative molecular mass and composition for Polygonatum sibiricum polysaccharide by high performance liquid chromatography]. AB - Polygonatum sibiricum Redoute (PSR) is a traditional Chinese medicine and Mongolia herbal medicine growing in Inner Mongolia. Polygonatum Sibiricum Polysaccharide (PSP) was isolated and purified with boiling water extraction, alcohol precipitation from the wild species roots of PSR. Proteins was removed from the crude PSP by methods of trypsin hydrolysate and Sevag. Purity, relative molecular mass and relative molecular mass distribution of PSP were determined using high performance liquid chromatography (HPLC). The buffer solution of NaH2PO4-Na2HPO4 (0.02 mol/L) was used as mobile phase, and the PSP was separated on a Shodex GPC column as a single symmetrical peak corresponding to a number mean molecular mass (Mn) of 7073 and relative molecular mass (Mr) of 7247, and polydispersity is 1.025. The sugar components of the PSP was analysed by HPLC and 13C NMR. Using water as mobile phase, the components of all acid hydrolysis of the PSP were separated on a Shodex SP0810 column. Glucose, galactose, mannose and fructose were used as the standard samples. The result showed that PSP is composed of fructose only. PMID- 16250451 TI - [Qualification and quantification of 10 sulfonamides in animal feedstuff by high performance liquid chromatography-electrospray tandem mass spectrometry]. AB - The presence of sulfonamide (SA) residues in foods is largely due to the raising of animals with sulfonamide antibiotics added or polluted feedstuff. Because of interference from the matrices, the commonly used immunoassay or chromatographic method is not suitable for the analysis of multi-SAs in feedstuff. A high performance liquid chromatographic-electrospray tandem mass spectrometric (HPLC/ESI-MS-MS) method has been established for the simultaneous determination of multi-SAs including sulfadiazine (SD), sulfapyridine (SPD), sulfamerazine (SM1), sulfameter (SM), sulfamethazine (SM2), sulfamethoxypyridazine (SMP), sulfamethoxazole (SMZ), sulfamonomethoxine (SMM), sulfadimethoxine (SDM) and sulfaquinoxaline (SQX). After solvent extraction, solid phase extraction, dilution and reversed-phase HPLC separation, SAs were detected by ESI-MS-MS under multi-reaction monitoring mode. The qualification analysis was done by using retention time and distribution of diagnostic ion pairs, and the quantification was based on the peak intensity of common fragment ion m/z 156. The limits of quantification for 10 SAs were 0.5 - 2.0 microg/kg (S/N = 10). The correlation coefficient of linear calibration curve was over 0.9995 within the SAs concentration range 2.0 - 200 microg/L except for SDM and SQX. At the spiked level of 1.0 mg/kg, the average recoveries for the 10 SAs were between 70% and 92%, the relative standard deviations were under 10% for intra-day and under 15% for inter-day. Routine tests showed the method was fast, sensitive, specific, and practical for the SAs determination in feedstuff. PMID- 16250452 TI - [Simultaneous determination of four fluoroquinolone residues in fish by ion-pair high performance liquid chromatography]. AB - An ion-pair high performance liquid chromatographic method with a fluorescence detector was established to determine four fluoroquinolone residues in fish. The chromatographic conditions were as follows: Waters microBondapak C18 column (3.9 mm i.d. x 300 mm, 10 microm) was used at 40 degres C with 11 mmol/L tetrabutylammonium bromide solution (pH 3.0) acetonitrile (94: 6, v/v) as the mobile phase with a flow rate of 1.0 mL/min, at the excitation wavelength of 280 nm and emission wavelength of 460 nm. The detection limit for fish was 1 microg/kg. The linear range was from 6-100 microg/kg, and correlation coefficients were more than 0.9995. At the levels of 10, 50 and 100 microg/kg, the recoveries were 76%-100% with relative standard deviations of less than 7%. The method developed can meet the requirement for residue analysis. PMID- 16250453 TI - [Simultaneous determination of seventeen sulfonamide residues in chickens by liquid chromatography-electrospray tandem mass spectrometry]. AB - A liquid chromatography-electrospray tandem mass spectrometry (LC-MS/MS) for the simultaneous determination of seventeen commonly used sulfonamide (SA) residues in chickens has been developed. Stable isotopic compound (13)C6-sulfamethazine was used as internal standard. Multi-reaction monitoring mode was employed for the quantitative determination. The separation was performed on a Capcell Pak C8 DD column (150 mm x 2.0 mm i.d., 5 microm) with a gradient system of water (containing 0.2% formic acid)-methanol (containing 0.2% formic acid) as mobile phase at a flow rate of 0.2 mL/min. Samples were prepared by homogenizing the chicken, extracting with acetonitrile, defatting with n-hexane and cleaning-up with Sep-Pak Silica solid-phase extraction. The detection limits of 0.02 - 1 microg/kg proved to be much better than the previously reported ones. Average recoveries of seventeen SAs ( spiked at the levels of 1, 5, 10 microg/kg) ranged from 52.3% to 124.9%, with relative standard deviations between 1.0% and 17.6%. Intra-day and inter-day variations of the method were all with in the acceptable ranges. The results demonstrated that the method is simple, accurate and suitable for the identification and quantification of these sulfonamide residues in chickens. PMID- 16250454 TI - [Determination of amino acids in Isatis indigotica Fort by reversed-phase high performance liquid chromatography with pre-column derivatization]. AB - A reversed-phase high performance liquid chromatographic method with pre-column derivatization for the determination of amino acids, which were known as the characteristic constituents of Isatis indigotica Fort was established. The amino acids in alkalescence were derivatized with 2, 4-dinitro-fluorobenzene (DNFB). A reproducible method for simultaneous qualitative analysis of glutamic acid (Glu), argirine (Arg) and proline (Pro) and quantitative analysis of Arg and Pro in Isatis indigotica Fort has been established. NaAc buffer (pH 6.4) acetonitrile (85:15, v/v) as mobile phase and a Sinochrom ODS-BP column were used. The detector was operated at 360 nm. The linear regressions of the standard curves were determined for Arg and for Pro. The method was carried out over the range of 0. 627 - 5.016 microg for Arg and 0. 874 - 7.000 microg for Pro. The recoveries were 98.5% and 98.4% with the relative standard deviations of 2.5% and 2.3% for Arg and Pro respectively. The results indicate that among the three amino acids in Isatis indigotica Fort, Arg content was the highest, Pro the second and Glu the lowest. The method has good accuracy and repeatability and it can be used for the quality control of Isatis indigotica Fort. PMID- 16250455 TI - [Separation and purification of the main glucosinolate from rapeseeds]. AB - A procedure for the preparative isolation of 1-S-[(1Z)-3-hydroxy-1-[(sulfooxy) imino]-4-pentenyl]-1-thio-beta-D-glucopyanose, potassium salt (progoitrin) from Brassica oleracea is reported. The major steps in this procedure were: (1) extraction of glucosinolates with methanol from Brassica oleracea; (2) separation and purification of glucosinolates by chromatographic column on alumina support; and (3) a follow-up reversed-phase separation by octadecyl (C18) silica support yielding progoitrin as the main content of glucosinolate. The structure of progoitrin was identified on its physicochemical properties by ultraviolet absorption spectrometry, infrared absorption, 'H nuclear magnetic resonance spectroscopy, mass spectrometry and elemental analysis. Spectroscopic data of the sample isolated were in agreement with those reported in the literature. The purity of progoitrin obtained was determined to be 99% by high performance liquid chromatography. The simplicity of the separation and purification procedure for glucosinolates and the high purity of progoitrin isolated would make the method an important reference for future research on glucosinolates and a favorable technique for future development. PMID- 16250456 TI - [Resolution of carotenoid isomers in Lycium barbarum L. by heuristic evolving latent projection]. AB - Lycium barbarum L., a kind of traditional Chinese herb, is found to have bioactivities such as anticancer, antioxidant, hypoglycemic and immunological activities. In both in vitro and in vivo studies, the carotenoids were found to be a class of the effective compounds. The carotenlysoids in Lycium barbarum L. were separated by high perforance liquid chromatography with diode array detection (HPLC-DAD). Seven peaks were obtained by HPLC on a C18 column with acetonitrile-methylene chloride (60:40, v/v) as mobile phase at a flow-rate of 1.0 mL/min. Most of the peaks that had been validated as single peaks in the 2 dimensional chromatography were found to be overlapping peaks. The overlapping chromatographic peaks were resolved by chemometric method--Heuristic Evolving Latent Projection (HELP) based on 3-dimensional data. As an example, the chromatogram and UV spectra of 4 isomers were obtained by resolving an overlapping peak. These results showed that the combination of chemometric methods and modern analytical instruments provides an effective method for the analysis of complex systems such as isomers. PMID- 16250457 TI - [Characterization and identification of Ralstonia solanacearum by ion-exchange chromatography]. AB - Ralstonia solanacearum, a widely distributed and economically important plant pathogen, was characterized by ion-exchange chromatography (IEC). Ralstonia solanacearum was chromatographed on a TOYOPEARL SuperQ-650 C column (200 mm x 4. 6 mm i. d. ) with gradient elution by A (0.02 mol/L piperazidine-chlorhydric acid buffer (pH 8.0) ) and B (A + 1 mol/L NaCl). The pure culture of R. solanacearum was separated into three fractions on a SuperQ-650 C column. They were found all belong to R. solanacearum after the fractions were identified by other biochemical methods. Because of their ability to oxidize 3 disaccharides (lactose, maltose and cellobiose) and 3 hexose alcohols (mannitol, sorbitol and dulcitol), they are classified as biovar III of R. solanacearum. Once the mobility was scaled using microscope and the pathogenetic ability was measured with 2,3 ,4-triphenyltetrazolium chlorid (TTC) medium, the two fractions were in different states. The results are very important to elucidate the multi-states of R. solanacearum and the mechanism of R. solanacearum's pathogenetic mutation. PMID- 16250458 TI - [Classification of terpenes and terpene oxides in volatile oil of Fruit of Acanthopanax senticosus (Rupr. Et Maxim) Harms with gas chromatographic retention parameters]. AB - Gas chromatographic (GC) retention parameters (A and B values) for 24 volatile compounds in volatile oil of Acanthopanax senticosus (Rupr. Et Maxim) Harms were obtained from the retention times under five temperature-programming conditions with self-developed GC_AB software based on Levenberg-Marquardt method. The correlation analysis between A and B parameters of terpene and its oxides was carried out. Good linear relationships between A and B parameters were built for seven monoterpenes (C10H16) , nine sesquiterpenes (C15H24), three monoterpene oxides (C10HnO) and three sesquiterpene oxides (C15HnOx) at the same carbon number, respectively. But poor A-B linearity was built for the group formed from monoterpenes and monoterpene oxides or that formed from sesquiterpenes and sesquiterpene oxides at the same carbon number. At the same time the A-B relationships for monoterpenes, sesquiterpenes and their oxides are not collinear. Therefore, this discipline is applied in the assistant identification for terpenes with different carbon numbers and in the classification between terpenes and their oxides. It shows that GC retention parameters calculated from retention times under several temperature-programming conditions are useful for the classification of monoterpenes, sesquiterpenes and their oxides in the analysis of volatile oils from traditional Chinese herbs. PMID- 16250459 TI - [Quantitative evaluation of the variation of aroma harmony in processed fruit and vegetable juices with gas chromatographic data]. AB - To develop a quantitative evaluation model for the variation of aroma harmony in processed fruit and vegetable juices, gas chromatographic data from juice samples were summed up by mathematic modeling. Based on the original fruit and vegetable juices, the total change in volatile compounds expressed in term of percentage between the treated samples by various processes and the original juice, that is, the deviation of samples, are calculated. They were then used to describe the total change of aroma compounds in the fruit and vegetable juices before and after the processing. To compare the influences of different processes on aroma harmony in fruit and vegetable juices, the samples were analyzed by gas chromatography under the same conditions and the data were obtained by comparing the deviations of the samples. The lemon juices concentrated either by freeze concentration or by vacuum evaporation were compared against the original lemon juices. The results showed that the freeze-concentration well retained not only the absolute contents of aroma compounds but also the aroma harmony of natural lemons. PMID- 16250460 TI - [Resolution of clenbuterol hydrochloride enantiomers by thin-layer chromatography on silica gel impregnated with beta-cyclodextrin]. AB - The resolution of clenbuterol hydrochloride enantiomers was achieved by thin layer chromatography on silica gel GF254 plates impregnated w ith beta cyclodextrin. The effect of stereoselective auxiliary (acetonitrile and alcohol) was investigated. Resolution of clenbuterol hydrochloride enantiomers could be attained by using alcohols of butanol, 2-butanol or tert-butanol together with acetonitrile as developing solvent. The optimal conditions of resolution were determined as follows: a plate prepared with 15.00 g silica gel GF254 impregnated with 1.00 g beta-cyclodextrin, acetonitrile-2-butanol (20:80, v/v) as developing solvent and developed at room temperature. Under these conditions, Rf of the two isomers of clenbuterol hydrochloride enantiomers were 0.34 and 0.72 respectively. The resolution was 4.09 with baseline separation and the spots in chromatogram were almost of the same size. PMID- 16250461 TI - [Determination of praziquantel and propargite residues in fish tissues by high performance liquid chromatography with solid-phase extraction]. PMID- 16250462 TI - [Isolation and purification of bioactive compounds from fermentation broth of Paecilomyces militaris]. PMID- 16250463 TI - [Application of simultaneous pyrolysis methylation-gas chromatography to the identification of paint varieties]. PMID- 16250464 TI - Sexual behavior and selected health measures: men and women 15-44 years of age, United States, 2002. AB - OBJECTIVE: This report presents national estimates of several measures of sexual behavior among males and females 15-44 years of age in the United States in 2002, as collected in the 2002 National Survey of Family Growth (NSFG). These data are relevant to demographic and public health concerns, including fertility and sexually transmitted diseases among teenagers and adults. Data from the 2002 NSFG are compared with previous national surveys. METHODS: The 2002 NSFG was conducted by the Centers for Disease Control and Prevention's (CDC) National Center for Health Statistics (NCHS) and is based on in-person, face-to-face interviews with a national sample of 12,571 males and females in the household population of the United States. The measures of sexual behavior presented in this report were collected using Audio Computer-Assisted Self-Interviewing (ACASI), in which the respondent enters his or her own answers into a laptop computer without telling them to an interviewer. RESULTS: Among adults 25-44 years of age, 97 percent of men and 98 percent of women have had vaginal intercourse; 90 percent of men and 88 percent of women have had oral sex with an opposite-sex partner; and 40 percent of men and 35 percent of women have had anal sex with an opposite-sex partner. About 6.5 percent of men 25-44 years of age have had oral or anal sex with another man. Based on a differently worded question, 11 percent of women 25 44 years of age reported having had a sexual experience with another woman. The public health significance of the findings is described. PMID- 16250465 TI - The evolutionary genetics of canalization. AB - Evolutionary genetics has recently made enormous progress in understanding how genetic variation maps into phenotypic variation. However why some traits are phenotypically invariant despite apparent genetic and environmental changes has remained a major puzzle. In the 1940s, Conrad Hal Waddington coined the concept and term "canalization" to describe the robustness of phenotypes to perturbation; a similar concept was proposed by Waddington's contemporary Ivan Ivanovich Schmalhausen. This paper reviews what has been learned about canalization since Waddington. Canalization implies that a genotype's phenotype remains relatively invariant when individuals of a particular genotype are exposed to different environments (environmental canalization) or when individuals of the same single- or multilocus genotype differ in their genetic background (genetic canalization). Consequently, genetic canalization can be viewed as a particular kind of epistasis, and environmental canalization and phenotypic plasticity are two aspects of the same phenomenon. Canalization results in the accumulation of phenotypically cryptic genetic variation, which can be released after a "decanalizing" event. Thus, canalized genotypes maintain a cryptic potential for expressing particular phenotypes, which are only uncovered under particular decanalizing environmental or genetic conditions. Selection may then act on this newly released genetic variation. The accumulation of cryptic genetic variation by canalization may therefore increase evolvability at the population level by leading to phenotypic diversification under decanalizing conditions. On the other hand, under canalizing conditions, a major part of the segregating genetic variation may remain phenotypically cryptic; canalization may therefore, at least temporarily, constrain phenotypic evolution. Mechanistically, canalization can be understood in terms of transmission patterns, such as epistasis, pleiotropy, and genotype by environment interactions, and in terms of genetic redundancy, modularity, and emergent properties of gene networks and biochemical pathways. While different forms of selection can favor canalization, the requirements for its evolution are typically rather restrictive. Although there are several methods to detect canalization, there are still serious problems with unambiguously demonstrating canalization, particularly its adaptive value. PMID- 16250466 TI - The evolutionary origin and elaboration of sociality in the aculeate Hymenoptera: maternal effects, sib-social effects, and heterochrony. AB - We discuss the evolutionary origin and elaboration of sociality using an indirect genetic effects perspective. Indirect genetic effects models simultaneously consider zygotic genes, genes expressed in social partners (especially mothers and siblings), and the interactions between them. Incorporation of these diverse genetic effects should lead to more realistic models of social evolution. We first review haplodiploidy as a factor that promotes the evolution of eusociality. Social insect biologists have doubted the importance of relatedness asymmetry caused by haplodiploidy and focused on other predisposing factors such as maternal care. However; indirect effects theory shows that maternal care evolves more readily in haplodiploids, especially with inbreeding and despite multiple mating. Because extended maternal care is believed to be a precondition for the evolution of eusociality, the evolutionary bias towards maternal care in haplodiploids may result in a further bias towards eusociality in these groups. Next, we compare kin selection and parental manipulation and then briefly review additional hypotheses for the evolutionary origin of eusociality. We present a verbal model for the evolutionary origin and elaboration of sib-social care from maternal care based on the modification of the timing of expression of maternal care behaviors. Specifically, heterochrony genes cause maternal care behaviors to be expressed prereproductively towards siblings instead of postreproductively towards offspring. Our review demonstrates that both maternal effect genes (expressed in a parental manipulation manner) and direct effect zygotic genes (expressed in an offspring control manner) are likely involved in the evolution of eusociality. We conclude by describing theoretical and empirical advances with indirect genetic effects and sociogenomics, and we provide specific quantitative genetic and genomic predictions from our heterochrony model for the evolutionary origin and elaboration of eusociality. PMID- 16250467 TI - Electron scattering filter design for a single field rotational total skin irradiation. AB - The aim of radiotherapy treatment of cutaneous T-cell lymphoma is to irradiate the skin with an appropriately homogeneous dose distribution up to a few millimetres in depth. This can be achieved by applying one of the total skin electron irradiation techniques. An aluminium/polystyrene foam electron scattering filter was designed so that the incident beam is broadened and degraded sufficiently to achieve a mean dose uniformity in a rectangular field of 180 cm height and 40 cm width. This paper reports on the development and construction of the electron scattering filter for use with a Varian 2100C accelerator, without MLCs, with a dose uniformity, over a useful field dimension of 180 cm height and 40 cm width, of +/- 7% about the mean, and an x-ray contamination of less than 2.4% beyond a depth of 3 cm. PMID- 16250468 TI - Comparative evaluation of an II based and a flat panel based cardiovascular fluoroscopy system within a clinical environment. AB - The image quality and dose parameters from a 2004 Siemens Axiom Artis dBC cardiac biplane with flat panel detector were evaluated and compared to similar parameters evaluated for a 1977 Toshiba DPF 2000A biplane cardiac unit with a conventional image intensifier. Image quality assessment was performed with the Westmead test object; using solid water as a patient equivalent absorber. The patient dose comparison of the two systems is based on dose area product meter readings for 1512 patient cases recorded over 6 months following installation of the Siemens flat panel digital unit. The image quality results indicate that: (a) high contrast resolution was better with the digital flat panel unit, (b) low contrast resolution is similar between systems, and (c) the threshold contrast of the flat panel system is the same or inferior to that of the image intensifier system. Input dose to the surface of the flat panel detector showed a strong dependence on field size, similar to the behaviour of image intensifier system. For the most common clinical procedure--Left Heart Study via Judkins--the average total dose area product reading was 64.0 Gy-cm2 against 67.7 Gy-cm2 for the digital and conventional units respectively (p = 0.27) indicating no significant difference in dose performance between the two x-ray machines. PMID- 16250469 TI - Shift in absorbed dose for megavoltage photons when changing to TRS-398 in Australia. AB - Australian primary standards of air kerma and absorbed dose are realized in 60Co gamma rays. To calibrate the megavoltage photon beams from linear accelerators, radiotherapy centres have their ionization chamber calibrated in a 60Co beam and then use a protocol to transfer this calibration to the higher energy. The radiotherapy community is in the process of changing from the ACPSEM Protocol (Second Edition 1998) based on an air kerma calibration to the IAEA's TRS-398 Code of Practice, based on an absorbed dose to water calibration. To evaluate the shift in absorbed dose resulting from the new protocol, the absorbed dose should be determined using both protocols and compared. We present a formula for this shift which can be used to check the result. To use this formula the centre needs to measure a displacement correction and know the ratio of the air kerma to absorbed dose to water calibration factors at 60Co. We calculate the change they should expect by using the average ratio of the air kerma and absorbed dose to water calibration factors for NE2571 and NE2561 chambers, based on Australian standards, and by estimating the displacement correction from published depth dose data. We find the absorbed dose in a megavoltage photon beam to increase by between 0.1 and 0.6% for NE2571 chambers and between 0.7 and 1.1% for NE2561 chambers, for beams up to 35 MV. The dose measured using TRS-398 is always higher. PMID- 16250470 TI - Effects of internal and external scatter on the build-up characteristics of Monte Carlo calculated absorbed dose for electron irradiation. AB - The effects of internal and external scatter on surface, build-up and depth dose characteristics simulated by Monte Carlo code EGSnrc for varying field size and SSD for a 10 MeV monoenergetic electron beam with and without an accelerator model are extensively studied in this paper. In particular, sub-millimetre surface PDD was investigated. The percentage depth doses affected significantly by the external scatter show a larger build-up dose. A forward shifted Dmax depth and a sharper fall-off region compared to PDDs with only internal scatter considered. The surface dose with both internal and external scatter shows a marked decrease at 110 cm SSD, and then slight further changes with the increasing SSD since few external scattered particles from accelerator model can reach the phantom for large SSDs. The sharp PDD increase for the 5 cm x 5 cm field compared to other fields seen when only internal scatter is considered is significantly less when external scatter is also present. The effect of external scatter on surface PDD is more pronounced for large fields than small fields (5 cm x 5 cm field). PMID- 16250471 TI - Preliminary dose response study of a gel dosimeter using 2-Hydroxyethyl Methacrylate (HEMA). AB - In this work we present a gel dosimeter based on 2-Hydroxyethyl Methacrylate (HEMA). The gel dosimeter is manufactured in normal atmospheric oxygen (normoxic) and undergoes a measurable change after irradiation. The gel is shown to provide a signal to noise ratio of up to at least 35 and have a linear change in transverse relaxation rate up to 70 Gy when measured with magnetic resonance imaging. PMID- 16250472 TI - A method for benchmarking CT scanners. AB - This study involved the development of an objective method to compare the performance of five CT scanners for the purpose of benchmarking. The method used to assess the scanners was to determine the dose-normalised noise at a spatial resolution of 5.5 cm(-1). This gave a dose-normalised percent noise between 0.37% and 0.76%. The scanners were also assessed for radiation dose to patients undergoing abdomen and head CT examinations. Patients' dose-length product (DLP) for the abdomen clinical examinations varied from 305 to 685 mGy-cm, and for the head clinical examinations from 333 to 900 mGy-cm. The study results demonstrated that the comparison of dose and spatial resolution normalised percent noise levels is a useful method of comparing CT scanner performance. PMID- 16250473 TI - Measurement of CT scanner dose profiles in a filmless department. AB - The measurement of the FWHM of the slice thickness radiation dose profile of a CT scanner using a prototype low sensitivity CR imaging plate has been investigated, as an alternative to the traditional method using envelope-packed industrial film. Using a standard Agfa clinical CR system to acquire the image, the FWHM of the dose profile can be accurately measured using readily available Public Domain software. An Agfa 18 x 24 cm CR cassette gives a pixel pitch of 113.5 microm, but with interpolation, the measurement accuracy can be less than 1 pixel. For a nominal 10 mm collimation, 15 successive measurements of the FWHM using CR gave an average width of 10.00 mm with a standard deviation of 0.02 mm. This may be compared with 4 successive measurements using film and a dual exposure technique to define the optical density at half peak height, yielding an average width of 9.98 mm with a SD of 0.03 mm. This prototype NDT plate is not a commercial product, but a radiotherapy plate with a similar sensitivity is available commercially and should give similar results. PMID- 16250474 TI - Accuracy of an electromagnetic tracking device for measuring hip joint kinematics during gait: effects of metallic total hip replacement prosthesis, source-sensor distance and sensor orientation. AB - The present study sought to investigate the effects of source-sensor distance, sensor orientation and the effects of metallic total hip replacement (THR) prostheses on the accuracy of the 3Space Tracker System (3STS). Using a simulated hip joint, the angles measured by the 3STS with six different source-sensor distances and two source-sensor orientations were recorded. Then the angles measured in the absence and presence of three different THR prostheses were compared. Both source-sensor distance and sensor orientation affects the accuracy of the 3STS. Measurements were only affected by the presence of one type of prosthesis. The 3STS was equally reliable, but less accurate with source-sensor distances of more than 25 cm. The small angular error and insensitivity of this device to the presence of some metallic THR prostheses make it a useful measurement tool for gait studies performed before and after THR surgery. PMID- 16250475 TI - Effect of intravenous contrast on treatment planning system dose calculations in the lung. AB - Intravenous contrast-enhanced computed tomography is utilised in radiotherapy lung treatment planning to improve the delineation of the tumour volume and nodal areas. In the resultant CT images, the electron density is increased within the vascular structures of the lung and the overall density in the lung volume may also be increased. As yet, it is unclear whether the change in density affects the accuracy of dose calculations based on this CT data. Two investigations were undertaken. Firstly, contrast-enhancement was simulated using an anthropomorphic phantom. In the second investigation, bulk density corrections were performed in an existing patient dataset. In both investigations, treatment plans were generated using both pre- and post-contrast datasets. The numbers of monitor units calculated in each of the plans were compared, as were the resulting isodose curves, dose volume histograms and physical mean lung doses. The numbers of monitor units calculated from the contrast- and non contrast-enhanced datasets agreed within 2%. The isodose curves and dose volume histograms showed very minor differences in size and shape. With the introduction of contrast agent, the physical mean lung doses calculated remained below the limit recommended for an acceptable plan. These results indicate that the introduction of contrast agent has a minimal dosimetric impact upon lung cancer treatment plans. PMID- 16250476 TI - Development of an Australian secondary standard for the reference air kerma rate measurement of 125I seeds. AB - Calibration of a High-Dose-Rate 1000 Plus ionisation chamber and associated electrometer for the determination of reference air kerma rate for 125I brachytherapy seeds has been developed. Traceability was established from ionisation chamber measurements of an Amersham model 6711 125I seed for which a primary standard measurement of reference air kerma rate has been made by the National Institute of Standards and Technology. The status of an Australian secondary standard is to be realised by establishing legal verifying authority from the National Measurement Institute. The calibrated chamber thus provides a measurement from which similar chambers in hospitals may be calibrated. For quality assurance this result was compared with the calibration certificate supplied by the Accredited Dosimetry Calibration Laboratory of the University of Wisconsin, USA. PMID- 16250477 TI - Medical technology horizon scanning. AB - Horizon scanning is becoming particularly important in the medical industry, in the identification and evaluation of emerging technologies. This paper examines the role biomedical engineers may have in horizon scanning new medical technologies and considers whether this is a useful activity for biomedical engineers. A horizon scanning methodology for conducting studies of emerging medical technologies is introduced, consisting of the three main phases of (a) a systematic literature review, in which a set approach is taken to the gathering of information; (b) scanning for publications across a range of different sources; and (c) consideration of the literature in relation to fixed benchmarks to indicate the quality of published information and reported achievements. This methodology has been successfully applied by the authors in a horizon scanning study for the purpose of advising a Government agency on the status of remote patient monitoring technology. PMID- 16250478 TI - Siemens X-ray dosimeter: historical note. PMID- 16250479 TI - Using a needle-nose vise grip to remove Kirschner wires. AB - A needle-nose vise grip can be used to facilitate removal of Kirschner wires in an office setting. PMID- 16250480 TI - End of a journey? PMID- 16250481 TI - How to read the orthopedic literature. PMID- 16250482 TI - Flexible intramedullary nailing of pediatric unstable forearm fractures. AB - In the pediatric population, the majority of displaced 2-bone forearm fractures can be reduced and stabilized by conservative means. In this article, we report on a retrospective study of 23 patients with unstable or open forearm fractures treated with closed reduction and percutaneous stabilization with flexible titanium nails. All fractures healed without complication, and final functional results were excellent. PMID- 16250483 TI - Is kyphoplasty the standard of care for compression fractures in the spine, especially in the elderly? AB - Vertebral fractures, particularly if they are nonsymptomatic, are old, and have not produced kyphotic deformity, should be treated with conservative care, including appropriate medical management. Those fractures that are symptomatic, produce pain that persists over several weeks (4 out of 10 on the visual analog score), and have resulted in some kyphotic deformity should be treated with a vertebroplasty or kyphoplasty. Patients with kyphotic deformity of less than 20% can obtain pain relief from vertebroplasty that is comparable to pain relief from kyphoplasty. For patients with a greater degree of kyphosis, only kyphoplasty can offer the predictable restoration of height and realignment of the spine. Currently we treat all Colles' fractures, all hip fractures, and all tibia plateau fractures. The time has come to treat all symptomatic vertebral fractures. PMID- 16250484 TI - Management of distal humerus fractures. AB - Fractures of the distal humerus are complex injuries that can be effectively treated with open reduction and internal fixation (ORiF). Exposure of a complex intra-articular fracture may best be achieved through a posterior approach with osteotomy of the olecranon process. The ulnar nerve must be identified and protected, the articular surface must be reduced anatomically, and rigid fixation must be applied to both the medial and lateral columns of the distal humerus. Range of motion should be initiated as soon as possible postoperatively. Complications such as ulnar neuropathy, elbow stiffness, heterotopic ossification, and nonunion should be treated aggressively. Total elbow arthroplasty represents an effective option for fractures that cannot be treated with ORIF. PMID- 16250485 TI - Rates of refracture associated with external fixation in pediatric femur fractures. AB - We conducted a small retrospective study of rates of early complications associated with external fixation of pediatric femur fractures and compared rates at our institution with those reported in the literature. In our series of 22 patients, early complications included 12 pin-track infections (54.5%), 2 cases of loss of reduction (9.1%), 1 pin-track abscess (4.5%), and 1 refracture (4.5%). Overall rates (ours combined with those reported by other investigators) were 4.7% (34/719) for refractures and 33.1% (224/677) for pin-track infections. Factors that correlated with refractures were open fracture, bilateral fracture, and longer time in fixator. Factors with inconclusive correlations were fracture pattern, dynamization status, fixator type, pin size, and number of pins. PMID- 16250486 TI - Biomechanical investigation of optimal fixation of isolated talonavicular joint fusion. AB - Despite reports of high nonunion rates for isolated talonavicular fusion, this procedure may be indicated for some patients, including those for whom increased stability of the talonavicular joint in triple arthrodesis is needed. In the biomechanical cadaveric study reported here, we evaluated fixation methods used to provide optimal stability of talonavicular arthrodesis. A physiologic 3-point loading model was used to measure dorsal displacement of the navicular on the talus with 1 or 2 cannulated 4.5-mm screws across the talonavicular joint both with and without one 7.3-mm cannulated screw across the subtalar joint. Statistically significant differences in displacement under cyclic loading to one half body weight were found. Use of 2 talonavicular screws and no subtalar screw or 1 talonavicular screw plus 1 subtalar screw decreased the motion, as compared with use of 1 talonavicular screw and no subtalar screw. PMID- 16250487 TI - Calciphylaxis: have you seen this disease? PMID- 16250488 TI - Bone morphogenetic proteins for tendon injuries: an orthopedic primer. AB - Throughout the world, tendon injuries are a significant source of pain and disability. Most research on treating tendon injuries has been focused on developing new surgical techniques and perfecting existing ones. Recently, attention has turned to the biology of tendon healing as a means to improve the outcome of such injuries. If the biological milieu of the healing tendon can be altered to produce a stronger tendon, then clinical outcome can be improved. Use of bone morphogenetic proteins (BMPs) is one proposed method for improving the biology of tendon healing. BMPs 12, 13, and 14 have each been shown to improve tendon healing in animal models. In this article, I review BMP history, current research and future directions, and potential application in treating tendon injuries. PMID- 16250489 TI - Examining American bioethics: its problems and prospects. PMID- 16250490 TI - Bioethics, social class, and the sociological imagination. PMID- 16250491 TI - The soul of a new machine: bioethicists in the bureaucracy. PMID- 16250492 TI - Demagogues, firefighters, and window dressers: who are we and what should we be? PMID- 16250493 TI - One cheer for bioethics: engaging the moral experiences of patients and practitioners beyond the big decisions. PMID- 16250494 TI - Too early for global ethics? PMID- 16250496 TI - Bioethics and the culture wars. PMID- 16250495 TI - Developments in bioethics from the perspective of HIV/AIDS. PMID- 16250497 TI - CQ sources/bibliography. PMID- 16250498 TI - Baseball and bioethics. PMID- 16250499 TI - Physician-assisted draft evasion: civil disobedience, medicine, and war. PMID- 16250500 TI - Dissecting "discrimination". PMID- 16250501 TI - [Thirty years, a jumping-off point to the future]. PMID- 16250502 TI - [Electroencephalographic correlates of brain states during verbal learning:Part I. Characteristics of local synchronization of an EEG]. PMID- 16250503 TI - [Effect of a sound stimulus on postural responses]. PMID- 16250504 TI - [Formation of the system of visual perception in ontogenesis]. PMID- 16250505 TI - [Interhemispheric asymmetry, visual learning and invariant identification of images]. PMID- 16250506 TI - [Visual acuity and the crowding effect in 8- to 17-year-old schoolchildren]. PMID- 16250507 TI - [Psychophysiological mechanisms of the difficulties in learning to write]. PMID- 16250508 TI - [Slow potentials of the brain before saccades in response to visual stimuli in schizophrenic patients]. PMID- 16250509 TI - [Eye movements induced by changes of the internal representation of the body configuration ]. PMID- 16250510 TI - [Mechanism of the ''fast neurogenic component'' of the ventilatory response to the initiation of locomotor activity]. PMID- 16250511 TI - [Clinical physiology: the role in studying basic problems of the regulation of renal functions in humans]. PMID- 16250512 TI - [A complex multiparameter study of the human systemic reactions to a dosed exposure to hypoxia ]. PMID- 16250514 TI - [Increase of the allergic intolerance to prosthetic materials and local anesthetics: immunophysiology and laboratory diagnostics ]. PMID- 16250513 TI - [The role of the renin-aldosterone system in osmoregulatory reactions of healthy subjects in response to desmopressin]. PMID- 16250516 TI - [Psychophysiological characteristics of nuclear power station operator activity as a factor in accident rate]. PMID- 16250515 TI - [Physiological criteria in defining the standards for training and competition loads in elite sports]. PMID- 16250517 TI - RCM ethics advisory committee: who are we and what do we do? PMID- 16250518 TI - Maternal mortality crisis in Chad. PMID- 16250519 TI - Human cloning--ever closer. PMID- 16250520 TI - A day at the breech. PMID- 16250521 TI - Florence. . . The experience of becoming a mother in exile. PMID- 16250522 TI - Addressing teenage pregnancy in Lewisham. PMID- 16250523 TI - Healthcare watchdog reports to parliament. PMID- 16250524 TI - The maternity conveyor belt. PMID- 16250525 TI - Intrapartum and postpartum bladder care. PMID- 16250526 TI - Doing reflections and being in practice. PMID- 16250527 TI - Domestic violence issues. PMID- 16250528 TI - Domestic violence issues. PMID- 16250529 TI - A delivery suite triage area in a midwifery unit. PMID- 16250530 TI - "Are midwives valued for the job they do?". PMID- 16250531 TI - Will AfC end inequality for enrolled nurses? PMID- 16250532 TI - The view from my Ivory Tower is not too great. Brian Belle-Fortune--a voice from children's nursing. PMID- 16250533 TI - Feeling the strain. PMID- 16250534 TI - Combating workplace stress. PMID- 16250535 TI - Developments in addressing the organ donor shortage. AB - Non-heartbeating donation is not a new concept but it is one that has been out of vogue for many years. However, increasing shortage of organs for transplantation has led to its increased use as a viable source of organs. This article outlines the rationale for its re-emergence and the challenges it poses for health care professionals. PMID- 16250536 TI - Pre-eclampsia. PMID- 16250537 TI - Blood cells. Part one--Bone marrow. PMID- 16250539 TI - Evaluation of a model of interprofessional education. AB - Interprofessional education among health care professionals has been recommended as a way to improve the quality of services. This paper analyses the results of an evaluative study of a practitioner-led, interprofessional programme for preregistration health care students, the Trust-Based Education and Training Programme, developed by South West London and St George's Mental Health NHS Trust in collaboration with several local universities. PMID- 16250538 TI - Understanding the nature of cancer-related fatigue. AB - Fatigue is one of the most frequently reported symptoms of cancer and is also a consequence of cancer treatment. There are a variety of tools available to measure fatigue but investigations into effective management have been inconclusive. PMID- 16250540 TI - Metformin. PMID- 16250542 TI - A need to see the patient rather than their offence. Interview by Tash Shifrin. PMID- 16250541 TI - Food choices in primary care: a summary of the evidence. AB - One of the main challenges facing primary care health care professionals is trying to encourage people to change their diet and to help them maintain that change. There is evidence that primary care interventions can influence food choice and this article highlights the factors that influence food choice, focusing on those studies that are most relevant to nurses. PMID- 16250544 TI - Current management of small-bowel obstruction. AB - The most significant advances in the management of small-bowel obstruction are developments in imaging modalities available to assist in the diagnosis itself, as well as to possibly assist in the early identification of those cases requiring urgent operative decompression. The most marked of these have been in the use and interpretation of contrast-enhanced CT. This has decreased the use of barium studies and has largely supplanted ultrasound and magnetic resonance imaging in the management of these patients. Diagnostic and therapeutic laparoscopic techniques are also growing in both capability and popularity. Laparoscopic adhesiolysis and the adjuvant of bioresorbable membranes each hold promise but have yet to become established as standard treatment. Further progress is needed in the detection of early, reversible strangulation. As a consequence, the fundamentals of the surgical management of small-bowel obstruction have evolved little over the past 15 years. With our persistent inability to detect reversible ischemia, a substantial risk of progression to irreversible ischemia remains when surgery is delayed, particularly in the setting of suspected complete obstruction. PMID- 16250545 TI - Carotid endarterectomy versus carotid angioplasty and stenting: a critical appraisal. PMID- 16250546 TI - Laparoscopic liver resections. PMID- 16250547 TI - Role of reduced heart rate volatility in predicting death in trauma patients. PMID- 16250548 TI - Breast reconstruction: a comparison of autogenous and prosthetic techniques. PMID- 16250549 TI - Role of the interventional radiologist in the management of abdominal abscesses. PMID- 16250550 TI - Comparison of whole organ pancreas and isolated islet transplantation for type 1 diabetes. PMID- 16250551 TI - Effect of bariatric surgery on long-term mortality. PMID- 16250552 TI - Improving the quality of care in bariatric surgery: the volume and outcome relationship. PMID- 16250553 TI - The current management of gastrointestinal stromal tumors. PMID- 16250554 TI - Role of adjuvant therapy in the management of pancreatic cancer. PMID- 16250555 TI - The pharmacologic modulation of the hypermetabolic response to burns. AB - Patients with burns less than 40% TBSA do not have catabolism unless sepsis develops. Those with burns more than 40% TBSA always experience catabolism, which causes metabolic derangements that persist for at least 1 year after the injury in most body tissues. The accomplishments of the past decade have placed us in the midst of an exciting paradigm shift from what used to be a primary concern (ie, mortality) to areas that are more likely to enhance the quality of life of burn survivors. Modulating postburn hypermetabolism for the burned patient is of overwhelming importance in both the immediate care stage and the rehabilitative stage. Postburn hypermetabolism cannot be completely reversed but may be manipulated by nonpharmacologic and pharmacologic means. Early burn wound excision and complete wound closure, prevention of sepsis, the maintenance of thermal neutrality for the patient by elevation of the ambient temperature, and graded resistance exercises during convalescence are simple, highly effective primary treatment goals. Although the initial burn injury and sepsis-related complications principally determine the extent of the metabolic response in burn victims, obligatory activity, background- and procedural-related pain, and anxiety also greatly increase metabolic rates. Judicious maximal narcotic support, appropriate sedation, and supportive psychotherapy are mandatory if their effects are to be minimized. Several anabolic and anticatabolic agents are available for use during immediate care and rehabilitation. Exogenous, continuous low-dose insulin infusion, beta-blockade with propranolol, and the use of the synthetic testosterone analogue oxandrolone are the most cost-effective and least toxic therapies to date. These greatly assist therapeutic minimization of the loss of lean body mass and linear growth delay and are effective in burned patients with and without sepsis. Adverse effects, cost benefits, and the ease of administration and monitoring must be examined when considering the possibility of their use. PMID- 16250556 TI - Image-guided video-assisted thoracoscopic resection of small peripheral lung nodules. PMID- 16250557 TI - The current treatment of achalasia. PMID- 16250558 TI - Portal hypertension: the role of shunting procedures. PMID- 16250559 TI - Workup of the incidental liver lesion. PMID- 16250560 TI - Severe acute respiratory syndrome: what have we learned. PMID- 16250561 TI - The differentiation of benign and malignant thyroid nodules. PMID- 16250562 TI - Does volume help predict outcome in surgical disease? PMID- 16250563 TI - The role of minimally invasive surgery for pancreatic pathology. AB - The last decade has seen an increase in the application of minimally invasive surgical procedures to the management of pancreatic disease. The role of the minimally invasive surgical procedure in the staging of pancreatic malignancies appears to be one of selective use in patients at high risk of occult metastatic disease based on high-resolution CT imaging and clinical presentation. These patients can be spared the morbidity of a nontherapeutic laparotomy. Laparoscopic pancreatic surgery is an advanced laparoscopic procedure with a significant learning curve. It should be considered only by surgeons with extensive experience in open pancreatic surgery who possess advanced laparoscopic skills. Early reports suggest that laparoscopic pancreatic surgery can be accomplished with acceptable morbidity and mortality rates for the resection of small benign and low-grade malignant lesions in the body and tail of the pancreas, as well as for the internal drainage of pancreatic pseudocysts. Its role in the management of lesions in the head, neck, and uncinate process of the pancreas has yet to be determined. PMID- 16250564 TI - Please, just a cleaning. PMID- 16250565 TI - A study of 275 retrieved Branemark oral implants. AB - The aim of this report was to describe the bone tissue response to Branemark oral implants retrieved from patients. The material consisted of consecutively received Branemark threaded oral implants and related patient data provided by clinicians. The implant samples were processed into undecalcified sections for evaluation under the light microscope. The analysis demonstrated a lower percentage of bone-to-implant contact for the unloaded implants as compared to the loaded implants. When the threads were divided into four different regions, the loaded implants had a lower percentage of bone-contacting length at the thread top as compared to the other three regions. PMID- 16250566 TI - Immediate restoration of implants placed into fresh extraction sockets for single tooth replacement: a prospective clinical study. AB - The aim of the present clinical study was to evaluate the placement of transmucosal implants into fresh extraction sockets and their immediate restoration with temporary crowns. A series of 22 cases with a 12-month follow-up is presented. Twenty-two patients (15 women and 7 men; mean age 39 years) who needed a single tooth replaced because of vertical or horizontal root fracture, caries, endodontic lesions, or periodontal disease were treated with immediate postextraction implant placement. The implant was then restored with a screw retained prosthetic restoration within 24 hours. Radiographic assessments were made at baseline and 12 months after implant placement. Clinical parameters, such as plaque score, mucositis score, probing attachment level, mucosal margin position, variation of gingival level, and variation of papilla position, were also measured at baseline and after 12 months of follow-up. At 12 months, no implants had failed. Radiographic examination revealed mean bone resorption of 0.5 mm at 12 months compared to baseline. The mean variation of gingival level, compared to the neighboring teeth, was -0.75 mm. Probing attachment levels were 0.79, 0.45, and 0.54 mm at proximal, buccal, and lingual sites, respectively. The values for the mucosal margin position were 2.9, 2.2, and 2.4 mm at proximal, buccal, and lingual sites, respectively. Regarding variation of papilla position, according to Jemt's index, 27 papillae presented with a score of 2 (61%) and 17 with a score of 3 (39%). An examination of oral hygiene and peri-implant soft tissue conditions at the 12-month follow-up visit revealed an overall frequency of plaque-carrying implant surfaces of 13%. Furthermore, mucositis (score 2) was not observed at any of the peri-implant units. Primary implant stability did not significantly increase over time. The immediate restoration of dental implants placed into fresh extraction sockets was shown to be a safe and predictable procedure. The success rate and radiographic and clinical results were comparable to those obtained following the standard protocol. Within the limits of the present investigation, immediate restoration of single-tooth implants placed in fresh extraction sockets can be considered a valuable option to replace a missing tooth. However, long-term clinical trials are needed to confirm the present results. PMID- 16250567 TI - Complications with surgical procedures utilizing connective tissue grafts: a follow-up of 500 consecutively treated cases. AB - The goal of this study was to evaluate the incidence and severity of the complications that occur after connective tissue grafts are used. Five hundred consecutively treated patients, for whom connective tissue grafts were used for root coverage or gingival augmentation, were included in this study. Complications did occur, but the rates and intensities seemed clinically acceptable. There was no pain reported in 81.4% of the patients, no bleeding in 97.0% of the patients, no infection in 99.2% of the patients, and no swelling in 94.6% of the patients. None of the factors evaluated in this study were associated with a statistically significant increase in the rate or intensity of complications. These factors included: age, sex of patient, smoking status, purpose of the graft (ie, for root coverage or for gingival augmentation), size of the recipient area, and the location of the defect being treated. Based on the results of this study, the incidence and severity of complications seemed to be clinically acceptable. PMID- 16250568 TI - Evaluation of efficacy of enamel matrix derivative in the treatment of intrabony defects: a 24-month multicenter study. AB - The aim of this prospective multicenter controlled clinical study was to evaluate the efficacy of Emdogain (Biora), an enamel matrix derivative (EMD), when combined with surgical treatment of periodontal angular defects, as compared to surgery alone, for up to 24 months of follow-up. The study was performed at six Italian universities and 11 private practices. Patients with one-, two-, or three wall angular defects were enrolled if intrabony defect depth (IBD) was 4 mm or more and probing pocket depth (PPD) was at least 6 mm. They were randomly allocated to either test or control groups. The test group was treated by the simplified papilla preservation (SPP) flap plus Emdogain after root conditioning with ethylenediaminetetraacetic acid. The control group was treated by SPP alone. Plaque Index, Gingival Index, PPD, and periodontal attachment level (PAL) at surgical sites were assessed at the presurgical examination (baseline). IBD was measured intraoperatively after debridement. IBD was also evaluated with a computer-aided technique, from periapical radiographs. Plaque Index, Gingival Index, PPD, PAL, and IBD were assessed at 12 and 24 months postsurgery. Data were further divided in two subgroups according to baseline IBD (6 mm or less and more than 6 mm). The differences between each follow-up and baseline, and between groups at each follow-up, for the above parameters were evaluated by standard statistical methods. One hundred fifty-three patients were recruited, accounting for 195 intrabony defects: 83 patients (108 defects) and 70 patients (87 defects) were allocated to the test and control groups, respectively. All parameters were improved at both 12 and 24 months, compared to baseline in both groups. In the test group, IBD, PPD, and PAL at 12 months were significantly better than these parameters in the control group. The test subgroup with IBD of more than 6 mm at baseline displayed a better outcome when compared to the 6 mm or less IBD subgroup. No significant adverse events related to the use of Emdogain were reported. The use of EMD as an adjunct to periodontal surgery in the treatment of angular defects significantly enhanced the rate and degree of periodontal regeneration. The control group also displayed significant tissue regeneration, but at a slower rate compared to the Emdogain group. The surgical procedure itself, with its goal of maximum preservation of the regenerative potential of periodontal tissues, proved to be effective in the treatment of periodontal angular defects. Pockets with IBD greater than 6 mm showed major improvement when treated with Emdogain. PMID- 16250569 TI - New telescopic crown protocol for partially edentulous patients: report of 32 cases. AB - In some periodontal cases, biomechanical, esthetic, medical, or economic concerns make it difficult for clinicians to employ either traditional therapy or restoration with an implant-supported prosthesis. To avoid any compromise and ensure a good result in such cases, we chose a removable telescopic prosthesis on natural teeth. In this paper we present a new clinical and technical method for the telescopic prosthesis, and we show that this is a predictable, reliable solution that is easy to realize, inexpensive, and comfortable for patients. PMID- 16250570 TI - The papilla amplification flap: a surgical approach to narrow interproximal spaces in regenerative procedures. AB - A modified surgical approach to interproximal guided tissue regeneration procedures (GTR) was used when anatomic conditions at the defect-associated interdental area rendered papilla preservation techniques very difficult. The main goal of this study was to evaluate the effectiveness of the papilla amplification flap (PAF) in obtaining and maintaining primary soft tissue closure of the interdental space above nonresorbable, titanium-reinforced, expanded polytetrafluoroethylene (e-PTFE) membranes and to quantify the regenerative outcomes obtained using this procedure. Seventeen patients with one deep intrabony defect associated with a narrow interproximal space were selected for this case-series clinical study. The application of the PAF in combination with e PTFE membranes resulted in clinically and statistically highly significant gains in clinical attachment levels (4.7 +/- 1.4 mm) and reductions in probing pocket depth (6.3 +/- 1.3 mm) after 1 year. Primary soft tissue closure of the interdental space was obtained in 100% of cases after completion of the surgery and maintained in 65% of cases during the initial healing period (6 weeks). Results from the present study indicate that the PAF can be considered a suitable soft tissue surgical approach for GTR treatment of intrabony defects when papilla preservation techniques are not recommended because of unfavorable local anatomic conditions interproximally. PMID- 16250571 TI - An accurate, single, platform-level impression technique for the fabrication of prosthetic restorations on dental implants. AB - The possibility of restorative inaccuracies increases with the number of impressions taken and subsequent models poured. Herein we describe a unique, splinted, dual-arch pick-up impression technique using abutment components instead of transfer copings. In our experience, this permits the fabrication of a definitive restoration as a result of a single impression. PMID- 16250572 TI - Immediate loading of mandibular implants in compromised patients: preliminary results. AB - To minimize the number of implant surgical procedures, a total of 72 single stage, transgingival Frialoc implants (Friadent) were placed in the mandibles of 18 patients who required implant restoration. The patients' ability to undergo surgical implant treatment was restricted because of either poor general medical health or psychologic conditions. All implants were immediately loaded with a bar retained overdenture. Only 4 of the 72 immediately loaded Frialoc implants (in one patient) failed over a control period of 29 months. The cumulative implant survival rate was 94.4%. Apart from an initial increase in bone resorption at two posterior implants supporting a cantilever bar (the cantilevers were removed 13 months after delivery of the prosthesis) and serious lip swelling in one patient, there were no further serious surgical, prosthetic, or general medical problems during the treatment period. The initial results of the study indicate that the single-stage Frialoc System allows fast prosthetic mandibular restorations. Because there is only one operation, the risk of local or general health complications in compromised patients is reduced. PMID- 16250573 TI - Guided periodontal regeneration using bilayered collagen membranes and bovine bone mineral in fenestration defects in the canine. AB - This study was performed to evaluate the effect of deproteinized bovine porous bone mineral (BBM) and BBM-collagen (BBMC) used alone or in combination with a bilayer collagen membrane in guided periodontal regeneration. In 12 dogs, contralateral surgical circular fenestration defects 5 mm in diameter were produced at the midbuccal aspect of the alveolar bone in 24 maxillary canines. Bone, periodontal ligament, and cementum were completely removed. Experimental sites were filled with BBM or BBMC. Bilayered collagen membranes covered half the experimental sites (BBM+M and BBMC+M), and the other half were left uncovered. Control sites remained empty; half were covered with collagen membranes (cont+M) and the underlying space spontaneously filled with blood, and half were left uncovered (cont). Three months postsurgery, undecalcified sections were prepared. Measurements were made using a caliper on a projection microscope, and the surface area of new bone and BBM particles within the healed surgical defect was evaluated using the point-counting method. In the experimental defects, new cementum covered 31% to 67% of the exposed dentin, with a significant difference between defects covered with membranes and defects that were not covered (P < .05). New cementum in the control (unfilled) defects also differed significantly between covered and uncovered defects. New bone growth presented a pattern similar to the cementum. There was no statistical difference between defects treated with BBM and BBMC, within both covered and uncovered groups. There was less connective tissue in the covered defects than in the uncovered defects (P < .05). The defects were filled with new bone, new connective tissue/bone marrow, and bovine bone particles. New bone area fraction was 23.4% to 25.2% in defects filled with BBMC and BBM, respectively (P = NS). Bone fraction area in membrane covered defects ranged from 34.4% to 36.8% in experimental defects (P = NS). All membrane-treated defects showed higher values for bone area fraction in comparison to the uncovered control defects. Particle area fraction ranged between 17.4% and 26.2%, with only BBMC and BBM+M defects showing a statistically significant difference (P < .05). Defects filled with submembranous blood clot exhibited significantly more new cementum and bone regeneration than experimental defects filled with BBM or BBMC. Treatment of defects with BBM or BBMC showed similar influences on bone and cementum regeneration in fenestration periodontal defects. The presence or absence of bilayered collagen membranes was the predominant factor influencing bone and cementum regeneration. PMID- 16250574 TI - [Mental development of children in condition of chronic physical abuse: methodical aspect]. AB - Mental dysfunction of 130 children aged 0-14 years with a history of family physical abuse was studied using a number of known methods adapted for early aged children including an original scale of mental dysfunction assessment, which has been elaborated in Mental Health Research Center (Moscow). A battery of clinico psychopathological and clinico-psychological methods allowed comprehensive assessment of mental state, with 95% cases of mental dysfunction being found in the group studied. The most pronounced were depression spectrum disorders. PMID- 16250575 TI - [Paradoxical kinesis phenomenon in focal hand dystonia--writer's cramp]. AB - Paradoxical kinesis (PK) phenomenon and its variants, exerting a beneficial influence on dystonia dynamics, are described using self clinical examination of 57 writer's cramp patients. PK was found in all the patients independently of writer's cramp variant, duration and severity. The most frequent writing maneuvers were as follows: hand printed (100%), proximal arm muscles writing (82.5%), individually selected writing instrument (67.5-80%), unusual means (67.5 75%), writing imitation with unlike-pen object (70%), marked papers (52.5%). The beneficial influence of PK phenomenon on dystonia expression may be considered as one of the directions of writer's cramp rehabilitation. PMID- 16250576 TI - [Epidemiology, clinical features and betaserc therapy of vertigo in initial and reversible cerebrovascular pathology]. AB - The study comprised two sections: epidemiologic and clinical. The aim of the epidemiologic investigation was to determine prevalence of vertigo, including its mild cases, in patients with initial and reversible forms of cerebrovascular pathology. While screening an open population, 726 men and women aged 35-60 years have been examined using uniform program. Vertigo was detected in 30%, frequent and long-lasting one in 14.6% cases (8.1% men and 21.9% women, p < 0.001). A clinical section included a profound examination of 40 patients. Along with duplex scanning of major brain arteries, neurologic and otoneurologic examination of the patients, aucoustic stem evoked potentials have been registered. Marked dysfunctions of autonomic nervous system and changes in functioning of different vestibular analyzer regions were revealed. High efficacy of betaserk used during 2 months in dosage 16 mg three times a day was demonstrated by improvement of the patient's state in 97% cases. PMID- 16250577 TI - [Biochemical changes of red blood cells in children with cerebral palsy and other organic brain disorders]. AB - One hundred and twenty-five children with cerebral palsy (n = 108) and organic CNS lesions (n = 17) have been studied. A control group included 30 children without any neurological lesions. Lipids and phospholipids spectra, Ca, Mg- and Na, K-ATPases, intensity of red blood cells lipid peroxidation and erythrocyte free magnesium content have been determined. Children with cerebral palsy had a higher percentage of cholesterol ethers and final products of lipid peroxidation in red blood cells--Shiff bases and a decrease of intraerythrocytic free magnesium. Patients with atonic-astatic syndrome were characterized by a sharp increase of Na, K-ATPase activity accompanied by increased content of such phospholipids as cardiolipin and phosphatide acid as well as Shiff bases. In patients with organic CNS lesions, Shiff bases content was similar to that in the control group, intracellular magnesium also did not differ significantly from the controls, but cardiolipin, phosphatide acid, cholesterol ethers and triglycerol concentrations were reduced. All the patients with neurological disorders had an increased percentage of phosphatidilcholin in red blood cells. PMID- 16250578 TI - [Clinical polymorphism of progressive Erb-Roth muscular dystrophy]. PMID- 16250579 TI - [The additional differential and diagnostic criteria of craniocerebral injury severity in children]. PMID- 16250580 TI - [Efficacy of movalis in marked neurological manifestations of lumbar osteochondrosis]. PMID- 16250582 TI - [Interactions of antiepileptic drugs]. PMID- 16250581 TI - [Depression in patients with chronic dermatoses and its treatment with coaxil]. PMID- 16250583 TI - [Aggravation of seizures by antiepileptic drugs]. PMID- 16250584 TI - [Medical and social aspects of childhood epilepsy]. PMID- 16250585 TI - [Aggravation of epileptic attacks under anticonvulsants influence]. PMID- 16250586 TI - [Non-steroid anti-inflammatory drugs in the treatment of Alzheimer's disease]. PMID- 16250587 TI - [WHO European Ministerial Conference on Mental Health, "facing the challenges, building solutions" (Helsinki, Finland, 12-15 January 2005)]. PMID- 16250588 TI - [Mental Health Declaration for Europe "facing the challenges, building solution"]. PMID- 16250589 TI - [Comment to the article by A. P. Rachin and Ya. B. Yudelson "Chronic daily head ache in children" published in No 1, 2005]. PMID- 16250590 TI - [Angiogenesis and viral infections]. AB - The review summarizes the results obtained from a study of the mechanisms responsible for angiogenesis during embryogenesis and adulthood in various abnormalities and in infections caused by hepatitis C virus and herpes simplex virus. PMID- 16250591 TI - [Detection of the circulation of Crimean-Congo hemorrhagic fever virus in the piedmont steppes of the North Caucasus]. AB - The laboratory verified cases of Crimean-Congo hemorrhagic fever (CCHF) in the piedmont steppes of the North Caucasus (Malgobeksky District, Republic of Ingushetia) are first described. The source of the first infection was Ixodidae ticks; three subsequent sources were contacts with the bloody discharges from patients. CCHF virus genome was detected in the blood of the cattle from an epidemic focus and in the pools of the Ixodes ticks Haemaphysalis parva Neum., 1897 and Boophilus annulatus Say, 1821, taken from cattle. The problem of including the piedmont steppes of the North Caucasus into the CCHF nosological area is discussed. PMID- 16250592 TI - [Anti-CD4 autoimmunity in HIV-infected persons in Russia]. AB - The frequency of anti-CD4 antibodies was determined in the sera or plasma derived from the patients infected with HIV-1 belonging to different genetic subgroups. The anti-CD4-antibodies in a dilution of > or = 1:1000 were found in 14% of the patients infected with the gagA/envA virus characteristic for injectable drug users in East Europe. The frequency of autoimmune antibodies among the HIV infected patients with envB virus was substantially less (4.4%). Competitive ELISA using monoclonal antibodies to different CD4 domains demonstrated that irrespective of the viral genotype, the autoimmune epitope is located within the D4 or D3/D4 domains of CD4 receptor. PMID- 16250593 TI - [Serum level of soluble forms of membranous antigens of immune system cells in carriers of virus hepatitis G markers]. AB - The levels of soluble CD38 (sCD38), CD50 (sCD50), and CD95 (sCD95) antigens and HLA class I (sHLA-I) were determined in the serum samples from persons infected with hepatitis G virus (HGV). HGV monoinfection was accompanied by a rise in the serum content of sCD38, sCD50, and sCD95 antigens. The serum presence of HGV RNA and anti-E2 HGV antibodies was characterized by the normal content of sCD38 and sCD95 while the level of sCD50 was elevated and the serum content of sHLA-I was decreased. If the serum contained only anti-E2 HGV antibodies, the level of sCD50 remained increased 4-fold. It is suggested that the higher adhesion-inhibiting level of sCD50 is a reason of a weak immune response to HGV and hence of a long HGV persistence in the body. PMID- 16250594 TI - [Theoretical prediction of the antigenic epitopes of severe acute respiratory syndrome (SARS-CoV) proteins and evaluation of their diagnostic value]. AB - Analysis of the sequence of a number of proteins of the virus causing severe acute respiratory syndrome (SARS-CoV) was used to theoretically predict antigenic epitopes. Nine sequences, encoding for the predicted epitopes in spike, nucleocapside, and membranous SARS CoV proteins, were synthesized by polymyrase chain reaction and cloned into the pGEX 4-T2 vector. The resultant plasmids were expressed in the E. coil cells and purified by glutathione-Sepharose affinity chromatography. PMID- 16250595 TI - [Human recombinant antibodies to Ebola virus: preparation and characteristics]. AB - Human recombinant antibodies against a purified Ebola virus (EV) lysate were selected from a combinatorial library of scFv-antibodies using the phage display technique. Nine unique antibodies were identified after sequencing the Vh- and Vl genes encoding the selected antibodies. Solid-phase enzyme immunoassay (EIA) indicated that these antibodies were able to bind both inactivated and native EV. Immunoblotting showed that 6 antibodies identified nucleoprotein (NP), one antibody did VP24 and another antibody did VP40. One of the selected antibodies reacted with two EP proteins: VP24 and VP40. Solid-phase EIA demonstrated cross reactivity with Marburg virus (MAR) and defined VP24 MAR as a target protein for the antibody. PMID- 16250596 TI - [Interferon-inducing properties of dry birch bark extract and its effect on experimental infection caused by hepatitis C virus]. AB - The interferon-inducing activity of dry birch bark extract containing betulin and its effect on hepatitis C virus (HCV)-induced infection were studied in the cultured cells. All samples of the extract induced increased interferon (IFN) production when it was orally and intraperitoneally administered. The dose dependence of the production of IFN showed a direct relation of the extract to interferon induction. There is strong evidence for that the extract can decrease the antigenic and infective activity of HCV in both in vitro experiments and HCV induced infection in mice. The preventive effect of the extract was found in both the HCV-infected cell cultures and the HCV-infected mice. PMID- 16250597 TI - [The rimantadine sensitivity spectrum in influenza A viruses circulating in the 2002-2004 epidemic seasons]. AB - A total of 200 influenza A virus strains were studied. Among them there 32 strains of A(H1N1) and 84 strains of A(H3N2) from the 2002-2003 epidemic season and 84 strains of A(H3N2) from the 2003-2004 epidemic season. Most rimantadine sensitive strains whose infectivity and hemagglutinating activity were found to decrease by the drug given at a concentration of 0.5 microg/ml. In terms of infectivity and hemagglutinating activity, the number of rimantadine-resistant strains of A (H1N1) was equal to 9.0% for A(H1N1) influenza viruses and to 10.0 and 14.0% for A (H3N2) influenza viruses in different seasons, respectively. PMID- 16250598 TI - [The morphology of the micro-capsulated polyacrylic acid-based formulation of measles vaccine]. AB - The morphology and virus localization were studied in the microcapsulated measles vaccine formulation involving polyacrylic acid (PAA) copolymers as a matrix. Transmission electron microscopy and phosphotungsic staining at a pH value of 2 to 7 showed that the morphology of microparticles was related to the value of pH and to the concentration of a polymer in the matrix. In the neutral medium, the microcapsules had the sizes of 0.5 to 10 microm, which were optimal for transport through the intestinal wall when immunization was orally used. Immunocytochemistry revealed measles virus antigen within the microparticles. The specific activity of the microcapsulated formulation of measles virus was as high as 3.36 and 4.31 lg of TCD50/0.5 ml for the samples containing 1 and 0.1% polymer, respectively. The findings suggest that the microparticles of the vaccine contain live measles virus. PMID- 16250599 TI - [Interaction of the glycoprotein gB of Aujeszky's disease virus (Suid herpesvirus 1) with cellular heparan sulfates]. AB - The interaction of the glycoprotein gB of Aujesky's disease virus (ADV, Suid herpesvirus 1) with heparan sulfates of BHK-21 cells was studied. The study used the native glycoprotein gB purified from the virus envelope by affinity chromatography based on gB-specific monoclonal antibodies. The cellular binding of the glycoprotein gB was specific and dose-dependent. Heparin, a structural analogue of heparan sulfates, inhibited the cellular binding of gB. The herapinase treatment of cells also resulted in the inhibition of the cellular binding of gB. The purified glycoprotein gB bound to heparin-Sepharose and was specifically eluated by heparin. The denaturation of gB was followed by 70-80% decreases in its binding to Sepharose-immobilized heparin. The findings may lead to the conclusion that plasma cell membranous heparan sulfates are one of the glycoprotein gB receptors in ADV. At the same time the heparin-binding site of ADV gB is at least partially conformation-dependent. PMID- 16250601 TI - Different views, common ground. PMID- 16250600 TI - [The virogenic status of cultured continuous simian lymphoid cells and their immunophenotypical characteristics]. AB - Immunophenotyping of cultured continuous simian lymphoid cultures using a panel containing monoclonal antibodies to human B- and T-cell antigens has been carried out, by employing enzyme immunoassay and flow cytofluorometry. The test cultures showed a wide variety of cells containing B- and T-cell antigens with their varying expression. The cultures were also found to comprise lymphoblasts simultaneously containing B- and T-cell markers. Simian lymphotropic viruses, such as EBV-like and STLV-1 retrovirus, detected apart or simultaneously, have been verified by polymerase chain reaction. Whether there is a possible relationship between the type of cells in the culture and the type of their replicating virus(es) is discussed in the paper. PMID- 16250602 TI - Integrating ministerial leadership at Provena Health. PMID- 16250603 TI - Treasuring memories of mentors. PMID- 16250604 TI - A "second generation" of ministry leadership. AB - Catholic health care leaders differ from others in the field in that "they are expected to serve as Jesus served, teach as Jesus taught, and lead as Jesus led, in order to heal as Jesus healed." The Catholic health ministry today is led largely by laypeople-what might be called the "first generation" of lay leaders. This first generation was privileged in that it was tutored by and worked alongside women and men religious. Those religious are now mostly gone from the ministry, and that first generation of lay leaders will also be retiring in the not too distant future. Leadership will then pass to a "second generation," laypeople who have not worked alongside religious. How is this new generation to learn "to heal as Jesus healed"? Catholic Health East (CHE), Newtown Square, PA, has developed a program explicitly directed at the recruitment and development of second-generation leaders. In its efforts to fill a position, the system first assembles a preferred-candidate profile, based on 15 competencies, including seven core competencies. CHE then employs a recruitment process based on behavioral event interviewing. All involved stakeholders participate in the interviews. PMID- 16250605 TI - Maintaining prophetic cultures. AB - The Catholic health ministry was founded by "prophetic people," people who shared some of the qualities shown by biblical prophets. If it is to endure and prosper, Catholic health care must foster prophetic cultures-cultures that positively encourage the development of new leaders possessing prophetic qualities. The Scriptures, particularly the Hebrew Scriptures, eloquently describe the characteristics these new leaders will require. The chief qualities that will be needed are memory (of the Hebrew and Christian tradition), creative imagination, orientation toward the community, steadfastness in commitment, patience in adversity, humility, a sense of humor, and an ability to express lamentation. To foster prophetic cultures, sponsors and board members must perform certain actions. They must act hopefully, set high standards, and clarify four leadership functions: conserve the organization's mission, manage resources efficiently, empower associates, and encourage everyone involved to respond to problems creatively. PMID- 16250606 TI - From CEO to mission leader. PMID- 16250607 TI - The notion of grace. AB - Catholic Healthcare West (CHW), San Francisco, which either sponsors or participates in three separate leadership development programs, sees the formation of new ministry leadership as a matter of the first importance. For the past four years, CHW has participated in CHA's Ecclesiology and Spiritual Renewal Program for System Leaders, the annual pilgrimage to the Vatican City in which ministry executives, board members, and sponsors get an opportunity to learn about the church's institutional structure and immerse themselves in its spiritual atmosphere. In 2002 the system established its CHW Learning Institute, which offers all employees training in leadership, clinical, governance, and employee development. Among other things, the institute has developed CHW's Competency Standards for Leadership. In 2004 CHW, with four other Catholic health care systems in the western United States, created the Ministry Leadership Center, Sacramento. This spring, 49 CHW managers were among the students enrolled in the center's inaugural classes. Among other subjects, they studied the distinctive competencies-intellectual, affective, and spiritual-required to lead a health care ministry in its operations and governance. PMID- 16250608 TI - Cancer and genetic medicine. A medical view. PMID- 16250610 TI - The role of the Bishop. PMID- 16250609 TI - Cancer and genetic medicine. An ethical view. PMID- 16250611 TI - Rural clergy and holistic care. PMID- 16250612 TI - PACE programs for sisters. PMID- 16250613 TI - The power of the simplest gesture. PMID- 16250614 TI - Strategic planning for a financial turnaround. PMID- 16250616 TI - [Comments on eversion technique of carotid endarterectomy]. AB - The authors describe their own modification of carotid endarterectomy by eversion technique which they use in the case of combined stenosis and kinking of internal carotid artery. Complete division of internal carotid artery from the common carotid artery at the bulb is performed in an oblique fashion, following by excision of redundant wall. Added longitudinal incision of common carotid artery enables perfect removal of the atherosclerotic plaque. Proper posterior wall anastomosis of common and internal carotid artery in the bulb is carried out by one-way-up technique. This is an alternative to parachute technique and gives perfect view of operating field, thus decreasing the risk of technical mistake. Eversion technique of carotid endarterectomy represents an ideal operative technique in the case of carotid stenosis combined with kinking. PMID- 16250615 TI - [Laparoscopic resection of the abdominal aortic aneurysm]. AB - Some 500 aortoiliac aneurysms are mostly electively operated in the Czech Republic each year. Miniinvasive approaches already widely accepted in general surgery are gradually becoming more routine in vascular surgery as well. Better patient comfort hand in hand with reduced hospital stay represent the major advantages of these procedures. The authors present their initial experience with laparoscopic AAA resection with the use of their own modification of the transperitoneal-retroperitoneal approach. Ischemic preconditionning's techniques with regard to the expected prolonged aortic clamping time were used during surgery. PMID- 16250618 TI - [Duodenal stump closure--still a live problem. A case report]. AB - Included is a case study of duodenal peptic ulcer bleeding. The patient was treated by Finsterer-Bancroft modification of II-type gastric resection. An early reoperation was indicated by reason of duodenal stump leak. The situation solved by means of duodenostomy. The authors discuss causes of duodenal stump insufficiency. Possibilities of prevention of this complication - which are cited in literature - are mentioned. Duodenostomy is a legitimate optimal solution of duodenal stump suture insufficiency. PMID- 16250617 TI - [The liver splenosis in a patient following a procedure for the malignant seminoma]. AB - Splenosis is characterized by a presence of islets of the splenic tissue in the abdominal and, rarely, also in the thoracic cavity, most frequently as a consequence of the splenic injury. The authors describe a case of splenosis in a patient followed-up after surgery and radiotherapy for the testicular seminoma and who, 42 years ago, had splenectomy for polytrauma. With respect to the location of the splenosis in the hepatic region, the differential diagnosis was difficult, bearing in mind a possibility of the malignancy relaps. Although splenosis was highly suspected, based on the results of the examinations conducted, with respect to the preexisting malignancy, the tumor was excised. The histopathologist confirmed the diagnosis of splenosis. This case is rare in the literature, considering the time gap between the diagnosis of splenosis and the trauma. PMID- 16250619 TI - [Laparoscopic distal resection of the pancreas]. AB - During the last two years, reports on laparoscopic procedures of the pancreas have been on increase. Laparoscopic resection of the pancreatic cauda is indicated, primarily, for benign cystic lesions of the cauda of the pancreas and for neuroendocrine tumors of the pancreas (mainly insulinomas). We have not recorded any report on the above procedure in the Czech literature. Therefore, in our case review, we have described laparoscopic distal resection of the pancreas with splenectomy for a pseudopapillary tumor of the pancreas. PMID- 16250620 TI - [A contemporary situation in the antimicrobial treatment of the secondary peritonitis]. AB - Peritonitis infection is polymicrobial, with the following prevailing pathogens - gram-negative aerobic and anaerobic bacteria and anaerobic cocci. Treatment of the secondary peritonitides includes surgical management of the infectious focus, initial empiric antimicrobial therapy and intensive adjunctive therapy aimed to control any secondary and terciary disorders. MATERIALS, METHODS AND RESULTS: Based on the results of the actual sensitivity of each pathogen of the intraabdominal infections and on the results of the past restrospective clinical studies, recommendations for initial empiric antimicrobial therapy of the secondary peritonitis have been compiled. Sensitivity of aerobic pathogens to parenterally administered antibiotics in the secondary peritonitits was regulary recorded and the results were summarized in 2004. The minimum inhibition concentrations (MIC in mg/l for the anaerobic bacteria and cocci are presented, based on present data of the National Reference Laboratory for Anaerobic Bacteria. A comparative retrospective study of the antimicrobial treatment in the secondary peritonitis from perforation of the diverticle of the large intestine, proved comparable results between treatment with antimicrobial monotherapy (piperacillin/tazobactam) and combined therapy (aminoglycosides with lincosamides or with nitroimidasoles). The effect of antimicrobial prophylaxis and therapy (aminopenicillines with the beta-lactamase inhibitor) was assessed in another restrospective study in acute appendicitis. The recommendations for initial empiric antimicrobial therapy in the secondary peritonitis are based on clinical and peroperative findings, on an estimated prevalence of the pathogens in the infectious focus and on sensitivity results of the individual antimicrobial medicines to the commonest pathogens in the secondary peritonitis. RESULTS: 1. Initial empiric antimicrobial treatment in the secondary peritonitis as a monotherapy with a broad-spectrum antibiotic or a combination of two antibiotics, should be effective against gram-negative aerobic, anaerobic bacteria and cocci. 2. With respect to morbidity and mortality rates, related to enterococcus infection not affected by antimicrobial therapy, administration of an antibiotic effective against enterococci is justified. 3. The choice of an antibiotic, duration and method of administration is based on the diagnosed infectious focus, peritonitis stage, on the current sensitivity findings of the commonest pathogens and on the overall condition of the patient (underlying disorders, immunodeficiency). 4. Treatment of other pathogens - Pseudomonas aeruginosa and mycoses is based on their laboratory confirmation and their sensitivity examination and usually is not an essential part of the initial empiric treatment. 5. In the initial empiric treatmet of early stages of the peritonitis, aminopenicilllin with the beta-lactamase inhibitor is the antibiotic of choice. 6. Due to its broad-spectrum effect against isolated bacteria and the safety parameters and low toxicity when used as monotherapy, pipracillin/tazobactam is indicated as the antibiotic of choice in advanced peritonitides with underlying disorders of the aboral part of the digestive tract. PMID- 16250621 TI - [Benefits of the computer navigation for reconstruction of the anterior crutiate ligament of the knee joint]. AB - AIM: The aim of this work is a radiological assessment of accuracy of the osseous channels location in initial reconstructions of the anterior crutiate ligament (PZV) conducted using a navigation system. METHODOLOGY: Following arthroscopic preparation of the joint space and registration of the essential data, conducting wires for channels in tibia and femur are navigated using the navigation system. The lig. patellae graft is then pulled-through and fixed using interference screws. The channels location was assessed using radiological methods according to Bernard-Hertel and Harner. RESULTS: A correct emergence of the femoral channel was found in 13 (87%) and of the tibial channel in 14 (93%) cases. The optimum graft location with respect to the both osseous channels was detected in 12 knee joints (80%). CONCLUSIONS: Exact location of the osseous channels is a principal prerequisite for a good result of the anterior crutiate ligament reconstruction. In case of standard procedures, this may be possible only in case the operating surgeon conducts a large number of reconstructions every year. In daily practice, however, 70% of reconstructions are conducted by less experienced orthopaedic surgeons. Our initial results indicate that the navigation system could become an effective tool for the surgeons who do not have the opportunity for conducting the above procedures too often. PMID- 16250622 TI - Ischemic colitis. PMID- 16250623 TI - [On experimental surgery]. PMID- 16250624 TI - Zeitgeist leadership. AB - Companies and leaders don't succeed or fail in a vacuum. When it comes to longterm success, the ability to understand and adapt to changing business conditions is at least as important as any particular personality trait or competency. A clear picture of how powerful the zeitgeist can be emerges from the authors' comprehensive study of the way the business landscape in the United States evolved, decade by decade, throughout the twentieth century. Six contextual factors in particular, they found, most affected the prospects for business: the level of government intervention in business, global events, demographics, shifts in social mores, developments in technology, and the strength or weakness of the labor movement. A lack of contextual sensitivity can trip up even the most brilliant executive. No less a luminary than Alfred P. Sloan was relieved of GM's day-to-day management in the 1930s because he was unwilling to meet with the new UAW. Conversely, an understanding of the zeitgeist can play a crucial but unheralded role in business performance. Jack Welch is widely credited with GE's remarkable success during the 1980s and 1990s, for example, but far less attention has been paid to his predecessor, the statesmanlike and prudent Reginald Jones, who sustained strong revenue and profit growth during the heavily regulated stagflation of the 1970s. To better understand this connection between business performance and context, the authors studied 1,000 great U.S. business leaders of the twentieth century and identified three distinct archetypes: Entrepreneurs, often ahead of their time, overcame dire challenges to build something new. Managers excelled at reading and exploiting the existing zeitgeist to grow their businesses. Leaders defied context to identify latent potential in businesses others considered mature, stagnant, or in decline. In every decade, all three archetypes were vital. It is the ongoing regeneration of this pattern in the business life cycle that ultimately sustains development and progress. PMID- 16250625 TI - Growing talent as if your business depended on it. AB - Traditionally, corporate boards have left leadership planning and development very much up to their CEOs and human resources departments-primarily because they don't perceive that a lack of leadership development in their companies poses the same kind of threat that accounting blunders or missed earnings do. That's a shortsighted view, the authors argue. Companies whose boards and senior executives fail to prioritize succession planning and leadership development end up experiencing a steady attrition in talent and becoming extremely vulnerable when they have to cope with inevitable upheavals- integrating an acquired company with a different operating style and culture, for instance, or reexamining basic operating assumptions when a competitor with a leaner cost structure emerges. Firms that haven't focused on their systems for building their bench strength will probably make wrong decisions in these situations. In this article, the authors explain what makes a successful leadership development program, based on their research over the past few years with companies in a range of industries. They describe how several forward-thinking companies (Tyson Foods, Starbucks, and Mellon Financial, in particular) are implementing smart, integrated, talent development initiatives. A leadership development program should not comprise stand-alone, ad hoc activities coordinated by the human resources department, the authors say. A company's leadership development processes should align with strategic priorities. From the board of directors on down, senior executives should be deeply involved in finding and growing talent, and line managers should be evaluated and promoted expressly for their contributions to the organization wide effort. HR should be allowed to create development tools and facilitate their use, but the business units should take responsibility for development activities, and the board should ultimately oversee the whole system. PMID- 16250626 TI - The office of strategy management. AB - There is a disconnect in most companies between strategy formulation and strategy execution. On average, 95% of a company's employees are unaware of, or do not understand, its strategy. If employees are unaware of the strategy, they surely cannot help the organization implement it effectively. It doesn't have to be like this. For the past 15 years, the authors have studied companies that achieved performance breakthroughs by adopting the Balanced Scorecard and its associated tools to help them better communicate strategy to their employees and to guide and monitor the execution of that strategy. Some companies, of course, have achieved better, longer-lasting improvements than others. The organizations that have managed to sustain their strategic focus have typically established a new corporate-level unit to oversee all activities related to strategy: an office of strategy management (OS M). The OSM, in effect, acts as the CEO's chief of staff. It coordinates an array of tasks: communicating corporate strategy; ensuring that enterprise-level plans are translated into the plans of the various units and departments; executing strategic initiatives to deliver on the grand design; aligning employees' plans for competency development with strategic objectives; and testing and adapting the strategy to stay abreast of the competition. The OSM does not do all the work, but it facilitates the processes so that strategy is executed in an integrated fashion across the enterprise. Although the companies that Kaplan and Norton studied use the Balanced Scorecard as the framework for their strategy management systems, the authors say the lessons of the OSM are applicable even to companies that do not use it. PMID- 16250627 TI - The passive-aggressive organization. AB - Passive-aggressive organizations are friendly places to work: People are congenial, conflict is rare, and consensus is easy to reach. But, at the end of the day, even the best proposals fail to gain traction, and a company can go nowhere so imperturbably that it's easy to pretend everything is fine. Such companies are not necessarily saddled with mulishly passive-aggressive employees. Rather, they are filled with mostly well-intentioned people who are the victirms of flawed processes and policies. Commonly, a growing company's halfhearted or poorly thought-out attempts to decentralize give rise to multiple layers of managers, whose authority for making decisions becomes increasingly unclear. Some managers, as a result, hang back, while others won't own up to the calls they've made, inviting colleagues to second-guess or overturn the decisions. In such organizations, information does not circulate freely, and that makes it difficult for workers to understand the impact of their actions on company performance and for managers to correctly appraise employees' value to the organization. A failure to accurately match incentives to performance stifles initiative, and people do just enough to get by. Breaking free from this pattern is hard; a long history of seeing corporate initiatives ignored and then fade away tends to make people cynical. Often it's best to bring in an outsider to signal that this time things will be different. He or she will need to address every obstacle all at once: clarify decision rights; see to it that decisions stick; and reward people for sharing information and adding value, not for successfully negotiating corporate politics. If those steps are not taken, it's only a matter of time before the diseased elements of a passive-aggressive organization overwhelm the remaining healthy ones and drive the company into financial distress. PMID- 16250628 TI - Information technology and the board of directors. AB - Ever since the Y2K scare, boards have grown increasingly nervous about corporate dependence on information technology. Since then, computer crashes, denial of service attacks, competitive pressures, and the need to automate compliance with government regulations have heightened board sensitivity to IT risk. Unfortunately, most boards remain largely in the dark when it comes to IT spending and strategy, despite the fact that corporate information assets can account for more than 50% of capital spending. A lack of board oversight for IT activities is dangerous, the authors say. It puts firms at risk in the same way that failing to audit their books would. Companies that have established board level IT governance committees are better able to control IT project costs and carve out competitive advantage. But there is no one-size-fits-all model for board supervision of a company's IT operations. The correct approach depends on what strategic "mode" a company is in whether its operations are extremely dependent on IT or not, and whether or not it relies heavily on keeping up with the latest technologies. This article spells out the conditions under which boards need to change their level of involvement in IT decisions, explaining how members can recognize their firms' IT risks and decide whether they should pursue more aggressive IT governance. The authors delineate what an IT governance committee should look like in terms of charter, membership, duties, and overall agenda. They also offer recommendations for developing IT policies that take into account an organization's operational and strategic needs and suggest what to do when those needs change. Given the dizzying pace of change in the world of IT, boards can't afford to ignore the state of their IT systems and capabilities. Appropriate board governance can go a long way toward helping a company avoid unnecessary risk and improve its competitive position. PMID- 16250629 TI - The hard side of change management. AB - Everyone agrees that managing change is tough, but few can agree on how to do it. Most experts are obsessed with "soft" issues, such as culture and motivation, but, say the authors, focusing on these issues alone won't bring about change. Companies also need to consider the hard factors-like the time it takes to complete a change initiative, the number of people required to execute it, and so forth. When the authors studied change initiatives at 225 companies, they found a consistent correlation between the outcomes of change programs (success versus failure) and four hard factors, which they called DICE: project duration, particularly the time between project reviews; integrity of performance, or the capabilities of project teams; the level of commitment of senior executives and staff; and the additional effort required of employees directly affected by the change. The DICE framework is a simple formula for calculating how well a company is implementing, or will be able to implement, its change initiatives. The framework comprises a set of simple questions that help executives score their projects on each of the four factors; the lower the score, the more likely the project will succeed. Companies can use DICE assessments to force conversations a bout projects, to gauge whether projects are on track or in trouble, and to manage project portfolios. The authors have used these four factors to predict the outcomes and guide the execution of more than 1,000 change management programs worldwide. Not only has the correlation held, but no other factors (or combination of factors) have predicted outcomes as successfully. PMID- 16250630 TI - Master of the house: Why a company should take control of its building projects. AB - When you head up a big construction project for your organization, coming in on time and on budget isn't enough. If you want to avoid squandering what is probably your company's largest capital investment, it's important to create a building that reflects your company's mission and produces a truly energizing work environment, says David Thurm, CIO of the New York Times Company and head of the team responsible for designing and building the Times' new corporate headquarters in Manhattan. The only way to get this kind of package-great design and innovative features that together further your business goals- is to take an active role. Assemble the right team, and then stay involved, asking hard questions about things that are generally taken as givens. Articulate a vision of your future work space, and drive the search for ways to realize this vision. In short, be a builder, not merely an owner. It's easy to understand why this approach is the exception rather than the rule. To most companies, design and construction seem foreign and forbidding, rife with pitfalls. Because of the murkiness of the field and a lack of experience and confidence, most companies play a relatively minor role in their construction projects. But it's a giant mistake to be a passive consumer when it comes to one of your most important assets. At best, you'll get well-intentioned guesses by others as to what you want; at worst, you'll end up with a building that's at odds with your identity. The author shares a series of lessons learned. Implicit in all of them: You have to push yourself as hard as you push your contractors. PMID- 16250631 TI - Four strategies for the age of smart services. AB - Most industrial manufacturers realize that the real money isn't in products but in services. Companies such as General Electric and IBM have famously made the transition: A large proportion of their revenues and margins come from providing value-added services to customers. But other companies attempting to do the same might miss the boat. It is not enough, the authors say, just to provide services. Businesses must now provide "smart services"--building intelligence (awareness and connectivity) into the products themselves. Citing examples such as Heidelberger Druckmaschinen's Internet-connected printing presses and Eaton Electrical's home-monitoring service, the authors demonstrate how a product that can report its status back to its maker represents an opportunity for the manufacturer to cultivate richer, longer-term relationships with customers. Four business models will emerge in this new, networked world. If you go it alone, it may be as an embedded innovator- that is, your networked product sends back information that can help you optimize service delivery, eliminate waste and inefficiency, and raise service margins. Or, you may pursue a more aggressive solutionist business model- that is, you position your networked product as a "complete solution provider," able to deliver a broader scope of high-value services than those provided by the embedded innovator's product. In the case of a system that aggregates and processes data from multiple products in a building or home, you may be either an aggregator or a synergist, partnering with others to pursue a smart-services opportunity. An aggregator's product is the hub, collecting and processing usage information- and creating a high-value body of data. A synergist's product is the spoke, contributing valuable data or functionality. Woe to the company that takes none of these paths; it'll soon find its former customers locked in--and happily--to other smart service providers. PMID- 16250632 TI - Design and synthesis of peptidomimetic severe acute respiratory syndrome chymotrypsin-like protease inhibitors. AB - Design, synthesis, and biological evaluation of peptidomimetic severe acute respiratory syndrome chymotrypsin-like protease (SARS-3CLpro) inhibitors for severe acute respiratory syndrome coronavirus (SARS-CoV) are described. These inhibitors exhibited antiviral activity against SARS-CoV in infected cells in the micromolar range. An X-ray crystal structure of our lead inhibitor (4) bound to SARS-3CLpro provided important drug-design templates for the design of small molecule inhibitors. PMID- 16250633 TI - A selective estrogen receptor modulator designed for the treatment of uterine leiomyoma with unique tissue specificity for uterus and ovaries in rats. AB - The design of a novel selective estrogen receptor modulator (SERM) for the potential treatment of uterine leiomyoma is described. 16 (LY2066948-HCl) binds with high affinity to estrogen receptors alpha and beta (ERalpha and ERbeta, respectively) and is a potent uterine antagonist with minimal effects on the ovaries as determined by serum biomarkers and histologic evaluation. PMID- 16250634 TI - 6-[1-(2,6-difluorophenyl)ethyl]pyrimidinones antagonize cell proliferation and induce cell differentiation by inhibiting (a nontelomeric) endogenous reverse transcriptase. AB - Two 2,6-difluoro-DABO derivatives (MC 1047, 1, and MC 1220, 2, respectively) were tested against endogenous, nontelomeric reverse transcriptase (endo-RT) in human differentiating cell systems to investigate their antiproliferative and cytodifferentiating activity. The two compounds significantly reduced cell proliferation and facilitated the morphological differentiation of cells. These results propose F(2)-DABOs as useful tools in preventive and/or curative therapy to counteract the loss of differentiation in dedifferentiating pathologies and as antiproliferative drugs in tumor therapy. PMID- 16250636 TI - Synthesis of a resveratrol analogue with high ceramide-mediated proapoptotic activity on human breast cancer cells. AB - Resveratrol, a natural product with a stilbene structure, exerts profound proapoptotic activity in human cancer cells, by triggering the accumulation of ceramide, a bioactive sphingolipid. We studied the biological effects of seven methoxylated and/or naphthalene-based resveratrol analogues and compared these compounds with resveratrol with the objective to identify an analogue with higher ceramide-mediated proapoptotic activity relative to resveratrol. Here we show that the compound with three hydroxyls and a naphthalene ring is the most effective in triggering apoptosis coupled to the induction of endogenous ceramide in human cancer cells. PMID- 16250635 TI - First-in-class pan caspase inhibitor developed for the treatment of liver disease. AB - A series of oxamyl dipeptides were optimized for pan caspase inhibition, anti apoptotic cellular activity and in vivo efficacy. This structure-activity relationship study focused on the P4 oxamides and warhead moieties. Primarily on the basis of in vitro data, inhibitors were selected for study in a murine model of alpha-Fas-induced liver injury. IDN-6556 (1) was further profiled in additional in vivo models and pharmacokinetic studies. This first-in-class caspase inhibitor is now the subject of two Phase II clinical trials, evaluating its safety and efficacy for use in liver disease. PMID- 16250637 TI - Lipid-soluble 3-pyridinol antioxidants spare alpha-tocopherol and do not efficiently mediate peroxidation of cholesterol esters in human low-density lipoprotein. AB - Because alpha-tocopherol (alpha-TOH) mediates the peroxidation of cholesterol esterified lipids in human low-density lipoprotein (LDL) in vitro and has displayed disappointing results against the onset and development of atherosclerosis, it may not be appropriate for use as a therapeutic in the prevention and/or treatment of the disease. Herein are described the experimental results demonstrating that 3-pyridinols spare alpha-TOH, do not efficiently mediate lipid peroxidation, and protect lipoprotein tryptophan residues in human LDL. PMID- 16250638 TI - Scaffold hopping with molecular field points: identification of a cholecystokinin 2 (CCK2) receptor pharmacophore and its use in the design of a prototypical series of pyrrole- and imidazole-based CCK2 antagonists. AB - A new molecular modeling approach has been used to derive a pharmacophore of the potent and selective cholecystokinin-2 (CCK(2)) receptor antagonist 5 (JB93182), based on features shared with two related series. The technique uses "field points" as simple and effective descriptions of the electrostatic and van der Waals maxima and minima surrounding a molecule equipped with XED (extended electron distribution) charges. Problems associated with the high levels of biliary elimination of 5 in vivo required us to design a compound with significantly lower molecular weight without sacrificing its nanomolar levels of in vitro activity. Two new series of compounds were designed to mimic the arrangement of field points present in the pharmacophore rather than its structural elements. In a formal sense, two of the three amides in 5 were replaced with either a simple pyrrole or imidazole, while some features thought to be essential for the high levels of in vitro activity of the parent compounds were retained and others deleted. These compounds maintained activity and selectivity for this receptor over CCK(1). In addition, the reduction in molecular weight coupled with lower polarities greatly reduced levels of biliary elimination associated with 5. This makes them good lead compounds for development of drug candidates whose structures are not obviously related to those of the parents and represents the first example of scaffold hopping using molecular field points. PMID- 16250639 TI - Optimization of the in vitro and in vivo properties of a novel series of 2,4,5 trisubstituted imidazoles as potent cholecystokinin-2 (CCK2) antagonists. AB - The systematic optimization of the structure of a novel 2,4,5-trisubstituted imidazole-based cholecystokinin-2 (CCK(2)) receptor antagonist afforded analogues with nanomolar receptor affinity. These compounds were now comparable in their potency to the bicyclic heteroaromatic-based compounds 5 (JB93182) and 6 (JB95008), from which the initial examples were designed using a field-point based molecular modeling approach. They were also orally active as judged by their inhibition of pentagastrin stimulated acid secretion in conscious dogs, in contrast to the bicyclic heteroaromatic-based compounds, which were ineffective because of biliary elimination. Increasing the hydrophilicity through replacement of a particular methylene group with an ether oxygen, as in 3-{[5-(adamantan-1 yloxymethyl)-2-cyclohexyl-1H-imidazole-4-carbonyl]amino}benzoic acid (53), had little effect on the receptor affinity but significantly increased the oral potency. Comparison of the plasma pharmacokinetics and the inhibition of pentagastrin-stimulated acid output following bolus intraduodenal administration of both 53 and 6 indicated that 53 was well absorbed, had a longer half-life, and was not subject to the elimination pathways of the earlier series. PMID- 16250640 TI - Molecular modeling and molecular dynamics simulation of the human A2B adenosine receptor. The study of the possible binding modes of the A2B receptor antagonists. AB - A molecular model of the human A(2B) adenosine receptor containing seven transmembrane alpha helices connected by three intracellular and three extracellular hydrophilic loops had been constructed. A molecular docking of seven structurally diverse xanthine antagonists of the A(2B) receptor was performed, and the differences in their binding modes were investigated. The 1 ns molecular dynamics (MD) simulations of several obtained ligand-receptor complexes inserted into the phospholipid bilayer were carried out. The conformational changes of the A(2B) receptor occurring during MD simulations were explored, and the stable binding modes of the studied antagonists were determined. According to the models presented in this work, the involvement of the His251, Asn282, Ser92, Thr89, and some aromatic residues in ligand recognition was determined. The obtained binding modes of the A(2B) antagonists demonstrate good agreement with the site-directed mutagenesis data. PMID- 16250641 TI - PostDOCK: a structural, empirical approach to scoring protein ligand complexes. AB - In this work we introduce a postprocessing filter (PostDOCK) that distinguishes true binding ligand-protein complexes from docking artifacts (that are created by DOCK 4.0.1). PostDOCK is a pattern recognition system that relies on (1) a database of complexes, (2) biochemical descriptors of those complexes, and (3) machine learning tools. We use the protein databank (PDB) as the structural database of complexes and create diverse training and validation sets from it based on the "families of structurally similar proteins" (FSSP) hierarchy. For the biochemical descriptors, we consider terms from the DOCK score, empirical scoring, and buried solvent accessible surface area. For the machine-learners, we use a random forest classifier and logistic regression. Our results were obtained on a test set of 44 structurally diverse protein targets. Our highest performing descriptor combinations obtained approximately 19-fold enrichment (39 of 44 binding complexes were correctly identified, while only allowing 2 of 44 decoy complexes), and our best overall accuracy was 92%. PMID- 16250642 TI - A new lead for nonpeptidic active-site-directed inhibitors of the severe acute respiratory syndrome coronavirus main protease discovered by a combination of screening and docking methods. AB - The coronavirus main protease, M(pro), is considered to be a major target for drugs suitable for combating coronavirus infections including severe acute respiratory syndrome (SARS). An HPLC-based screening of electrophilic compounds that was performed to identify potential M(pro) inhibitors revealed etacrynic acid tert-butylamide (6a) as an effective nonpeptidic inhibitor. Docking studies suggested a binding mode in which the phenyl ring acts as a spacer bridging the inhibitor's activated double bond and its hydrophobic tert-butyl moiety. The latter is supposed to fit into the S4 pocket of the target protease. Furthermore, these studies revealed etacrynic acid amide (6b) as a promising lead for nonpeptidic active-site-directed M(pro) inhibitors. In a fluorimetric enzyme assay using a novel fluorescence resonance energy transfer (FRET) pair labeled substrate, compound 6b showed a K(i) value of 35.3 muM. Since the novel lead compound does not target the S1', S1, and S2 subsites of the enzyme's substrate binding pockets, there is room for improvement that underlines the lead character of compound 6b. PMID- 16250643 TI - Pyrazolo[3,4-c]pyridazines as novel and selective inhibitors of cyclin-dependent kinases. AB - Pyrazolopyridazine 1a was identified in a high-throughput screening carried out by BASF Bioresearch Corp. (Worcester, MA) as a potent inhibitor of CDK1/cyclin B and shown to have selectivity for the CDK family. Analogues of the lead compound have been synthesized and their antitumor activities have been tested. A molecular model of the complex between the lead compound and the CDK2 ATP binding site has been built using a combination of conformational search and automated docking techniques. The stability of the resulting complex has been assessed by molecular dynamics simulations and the experimental results obtained for the synthesized analogues have been rationalized on the basis of the proposed binding mode for compound 1a. As a result of the SAR study, monofuryl 1o has been synthesized and is one of the most active compounds against CDK1 of this series. PMID- 16250644 TI - Synthesis, structure-activity relationships, and biological properties of 1 heteroaryl-4-[omega-(1H-indol-3-yl)alkyl]piperazines, novel potential antipsychotics combining potent dopamine D2 receptor antagonism with potent serotonin reuptake inhibition. AB - A series of novel bicyclic 1-heteroaryl-4-[omega-(1H-indol-3-yl)alkyl]piperazines was synthesized and evaluated on binding to dopamine D(2) receptors and serotonin reuptake sites. This class of compounds proved to be potent in vitro dopamine D(2) receptor antagonists and in addition were highly active as serotonin reuptake inhibitors. Some key representatives showed potent pharmacological in vivo activities after oral dosing in both the antagonism of apomorphine-induced climbing and the potentiation of 5-HTP-induced behavior in mice. On the basis of the preclinical data, 8-{4-[3-(5-fluoro-1H-indol-3-yl)propyl]piperazin-1-yl}-4H benzo[1,4]oxazin-(R)-2-methyl-3-one (45c, SLV314) was selected for clinical development. In vitro and in vivo studies revealed that 45c has favorable pharmacokinetic properties and a high CNS-plasma ratio. Molecular modeling studies showed that the bifunctional activity of 45c can be explained by its ability to adopt two different conformations fitting either the dopamine D(2) receptor pharmacophore or the serotonin transporter pharmacophore. PMID- 16250645 TI - Azepanone-based inhibitors of human cathepsin L. AB - The extension of a previously reported cathepsin K azepanone-based inhibitor template to the design and synthesis of potent and selective inhibitors of the homologous cysteine protease cathepsin L is detailed. Structure-activity studies examining the effect of inhibitor selectivity as a function of the P3 and P2 binding elements of the potent cathepsin K inhibitor 1 revealed that incorporation of either a P3 quinoline-8-carboxamide or a naphthylene-1 carboxamide led to increased selectivity for cathepsin L over cathepsin K. Substitution of the P2 leucine of 1 with either a phenylalanine or a beta naphthylalanine also resulted in an increased selectivity for cathepsin L over cathepsin K. Molecular modeling studies with the inhibitors docked within the active sites of both cathepsins L and K have rationalized the observed selectivities. Optimization of cathepsin L binding by the combination of the P3 naphthylene-1-carboxamide with the P2 beta-naphthylalanine provided 15, which is a potent, selective, and competitive inhibitor of human cathepsin L with a K(i) = 0.43 nM. PMID- 16250646 TI - Comparative epitope mapping with saturation transfer difference NMR of sialyl Lewis(a) compounds and derivatives bound to a monoclonal antibody. AB - The monoclonal antibody GSLA-2 recognizes the sialyl Lewis(a) (sLe(a)) epitope, which has an increased occurrence on mucin type glycoproteins of patients with colorectal carcinoma. GSLA-2 is therefore used in tumor diagnosis. To advance the understanding of this highly specific molecular recognition reaction, we have analyzed the binding epitope of sLe(a) at atomic resolution using saturation transfer difference NMR. To compare, the binding epitopes of sialyl Lewis(x) (sLe(x)) and of four synthetic derivatives of sLe(a) were explored. Surface plasmon resonance experiments furnished thermodynamic and kinetic data. It is observed that all pyranose rings of sLe(a) are in contact with the protein surface, with the neuramic acid residue receiving the largest fraction of saturation transfer. In contrast, sLe(x) binds very weakly, though specifically to GSLA-2, with a different binding epitope. This study provides a comprehensive picture of the recognition sLe(a) and explains the exquisite specificity of the antibody. PMID- 16250647 TI - 2-n-Butyl-9-methyl-8-[1,2,3]triazol-2-yl-9H-purin-6-ylamine and analogues as A2A adenosine receptor antagonists. Design, synthesis, and pharmacological characterization. AB - Two types of adenosine receptor ligands were designed, i.e., 9H-purine and 1H imidazo[4,5-c]pyridines, to obtain selective A(2A) antagonists, and we report here their synthesis and binding affinities for the four adenosine receptor subtypes A(1), A(2A), A(2B) and A(3). The design was carried out on the basis of the molecular modeling of a number of potent adenosine receptor antagonists described in the literature. Three compounds (25b-d) showed an interesting affinity and selectivity for the A(2A) subtype. One of them, i.e., ST1535 (2-n butyl-9-methyl-8-[1,2,3]triazol-2-yl-9H-purin-6-ylamine, 25b) (K(i) A(2A) = 6.6 nM, K(i) A(1)/A(2A) = 12; K(i) A(2B)/A(2A) = 58; K(i) A(3)/A(2A) > 160), was selected for in vivo study and shown to induce a dose-related increase in locomotor activity, suggestive of an A(2A) antagonist type of activity. PMID- 16250648 TI - Naphtho[1,2-d]isothiazole acetic acid derivatives as a novel class of selective aldose reductase inhibitors. AB - Acetic acid derivatives of naphtho[1,2-d]isothiazole (NiT) were synthesized and tested as novel aldose reductase (ALR2) inhibitors. The parent compound 11 exhibited a fair inhibitory activity (IC(50) = 10 muM), which was enhanced by 2 orders of magnitude by introducing a second carboxylic group at position 4 (13 and 14: IC(50) = 0.55 and 0.14 muM, respectively). Substitution of the acetic acid function with an apolar group gave inactive (29) or poorly active (25, 26, 30) compounds, thus demonstrating that the 2-acetic group is involved in the enzyme pharmacophoric recognition while the 4-carboxylic moiety has only an accessory role. The potent compounds 11, 13, 14, 26 all proved to be selective for ALR2, since none of them inhibited aldehyde reductase, sorbitol dehydrogenase, or glutathione reductase. The isopropyl ester 31, a prodrug of 14, was found to be effective in preventing cataract development in severely galactosemic rats, when administered as an eyedrop solution. The theoretical binding mode of 13 and 14, obtained by docking simulations into the ALR2 crystal structure, was fully consistent with the structure-activity relationships in the NiT series. PMID- 16250649 TI - Selective inhibitors of the serine protease plasmin: probing the S3 and S3' subsites using a combinatorial library. AB - A combinatorial library of 400 serine protease inhibitors with the general structure Cbz-X(aa)-Trp-cyclohexanone-Trp-Y(aa)-OH has been constructed. The library was synthesized on the solid phase using mix-and-split synthesis, where 20 different amino acids were incorporated at both the X(aa) and Y(aa) positions. These two positions correspond to the S3 and S3' subsites of the active site. Iterative deconvolution was used to identify hits from the library. The library was screened against four serine proteases: plasmin, kallikrein, thrombin, and trypsin. Seven inhibitors from the library that showed promising activities were resynthesized using solution-phase methods. Four of these compounds were good inhibitors of plasmin with IC(50) values in the range of 2.7-3.6 microM. The most potent of these inhibitors showed >150-fold selectivity for plasmin when compared to the other three serine proteases. PMID- 16250650 TI - Biospectra analysis: model proteome characterizations for linking molecular structure and biological response. AB - Establishing quantitative relationships between molecular structure and broad biological effects has been a long-standing goal in drug discovery. Evaluation of the capacity of molecules to modulate protein functions is a prerequisite for understanding the relationship between molecular structure and in vivo biological response. A particular challenge in these investigations is to derive quantitative measurements of a molecule's functional activity pattern across different proteins. Herein we describe an operationally simple probabilistic structure-activity relationship (SAR) approach, termed biospectra analysis, for identifying agonist and antagonist effect profiles of medicinal agents by using pattern similarity between biological activity spectra (biospectra) of molecules as the determinant. Accordingly, in vitro binding data (percent inhibition values of molecules determined at single high drug concentration in a battery of assays representing a cross section of the proteome) are useful for identifying functional effect profile similarity between medicinal agents. To illustrate this finding, the relationship between biospectra similarity of 24 molecules, identified by hierarchical clustering of a 1567 molecule dataset as being most closely aligned with the neurotransmitter dopamine, and their agonist or antagonist properties was probed. Distinguishing the results described in this study from those obtained with affinity-based methods, the observed association between biospectra and biological response profile similarity remains intact even upon removal of putative drug targets from the dataset (four dopaminergic [D1/D2/D3/D4] and two adrenergic [alpha1 and alpha2] receptors). These findings indicate that biospectra analysis provides an unbiased new tool for forecasting structure-response relationships and for translating broad biological effect information into chemical structure design. PMID- 16250651 TI - Solvent constraints on the property space of acetylcholine. 2. Ordered media. AB - The objective of this study was to investigate the conformational and property spaces of acetylcholine in hydrated octanol and in a membrane model. Molecular dynamics simulations of long duration (15 ns) were carried out, yielding 3000 conformers. For each, we calculated N(+)-C8 distance, solvent-accessible surface area (SAS), polar surface area (PSA), dipole moment, and lipophilicity (virtual logP). Their variations as a function of the dihedral angles tau(2) and tau(3) remained unexpectedly broad and comparable to those seen previously in a vacuum, in water, and in chloroform.(12) Thus, each of the seven conformational clusters was able to access a marked proportion of the lipophilicity space accessible to acetylcholine (0.40 in the logP scale). Histograms of logP distributions revealed two overlapping populations, namely more lipophilic and more hydrophilic. Their deconvolution into two Gaussian curves demonstrated solvent-mediated constraints on the lipophilicity space of acetylcholine, clearly showing how a polar medium favors polar conformers, whereas the opposite is true for media of low polarity. PMID- 16250652 TI - Toward selective histone deacetylase inhibitor design: homology modeling, docking studies, and molecular dynamics simulations of human class I histone deacetylases. AB - Histone deacetylases (HDACs) play an important role in gene transcription. Inhibitors of HDACs induce cell differentiation and suppress cell proliferation in tumor cells. Although many HDAC inhibitors have been designed and synthesized, selective inhibition for class I HDAC isoforms is a goal that has yet to be achieved. To understand the difference between class I HDAC isoforms that could be exploited for the design of isoform-specific HDAC inhibitors, we have built three-dimensional models of four class I histone deacetylases, HDAC1, HDAC2, HDAC3, and HDAC8. Comparison of the homology model of HDAC8 with the recently published X-ray structure shows excellent agreement and validates the approach. A series of HDAC inhibitors were docked to the homology models to understand the similarities and differences between the binding modes. Molecular dynamic simulations of these HDAC-inhibitor complexes indicate that the interaction between the protein surface and inhibitor is playing an important role; also some active site residues show some flexibility, which is usually not included in routine docking protocols. The implications of these results for the design of isoform-selective HDAC inhibitors are discussed. PMID- 16250653 TI - Is antagonism of E/Z-guggulsterone at the farnesoid X receptor mediated by a noncanonical binding site? A molecular modeling study. AB - Guggulsterone 1, the active principle of guggulipid, has been used in ethnic medicine for thousands of years for its antinflammatory and antilipidemic activities. The activities of 1 are apparently mediated by its interaction with an array of nuclear receptors, including endocrine steroid receptors and metabolic lipid receptors. Although relatively weak, the activity at the metabolic farnesoid X receptor (FXR) is particularly intriguing, as 1 is, so far, the only antagonist known for this receptor, with a peculiar ability of gene selective modulation. We report here a systematic study aimed at identifying the potential binding pocket of 1 at FXR. Although 1 could be docked into the canonical binding site, we identified a novel, so far undescribed binding pocket, localized near the loop region between helix 1 and helix 2. This novel binding pocket may explain some of the peculiar characteristics of 1 when acting at FXR. PMID- 16250655 TI - MetaSite: understanding metabolism in human cytochromes from the perspective of the chemist. AB - Identification of metabolic biotransformations can significantly affect the drug discovery process. Since bioavailability, activity, toxicity, distribution, and final elimination all depend on metabolic biotransformations, it would be extremely advantageous if this information could be produced early in the discovery phase. Once obtained, this information can help chemists to judge whether a potential candidate should be eliminated from the pipeline or modified to improve chemical stability or safety of new compounds. The use of in silico methods to predict the site of metabolism in phase I cytochrome-mediated reactions is a starting point in any metabolic pathway prediction. This paper presents a new method, specifically designed for chemists, that provides the cytochrome involved and the site of metabolism for any human cytochrome P450 (CYP) mediated reaction acting on new substrates. The methodology can be applied automatically to all the cytochromes for which 3D structure is known and can be used by chemists to detect positions that should be protected in order to avoid metabolic degradation or to check the suitability of a new scaffold or prodrug. The fully automated procedure is also a valuable new tool in early ADME-Tox assays (absorption, distribution, metabolism, and excretion toxicity assays), where drug safety and metabolic profile patterns must be evaluated as soon, and as early, as possible. PMID- 16250656 TI - Fast small molecule similarity searching with multiple alignment profiles of molecules represented in one-dimension. AB - Multiple sequence alignment has proven to be a powerful method for creating protein and DNA sequence alignment profiles. These profiles of protein families are useful tools for identifying conserved motifs, such as the catalytic triad of the serine protease family or the seven transmembrane helices of the G-protein coupled receptor family. Ultimately, the understanding of the critical motifs within a family is useful for identifying new members of the family. Due to the complexity of protein-ligand recognition, no universally accepted method exists for clustering small molecules into families with the same or similar biological activity. A combination of the concept of multiple sequence alignment and the 1 dimensional molecular representation described earlier offers a new method for profiling sets of small molecules with the same biological activity. These small molecule profiles can isolate key commonalities within the set of bioactive compounds much like a multiple sequence alignment can isolate critical motifs within a protein family. The small molecule profiles then make useful tools for searching small molecule databases for new compounds with the same biological activity. The technique is demonstrated here using the human ether-a-go-go potassium channel and the kinase SRC. PMID- 16250654 TI - 2,5-diketopiperazines as potent, selective, and orally bioavailable oxytocin antagonists. 2. Synthesis, chirality, and pharmacokinetics. AB - A short stereoselective synthesis of a series of chiral 7-aryl-2,5 diketopiperazines oxytocin antagonists is described. Varying the functionality and substitution pattern of substituents in the 7-aryl ring and varying the chirality of this exocyclic ring have produced potent oxytocin antagonists (pK(i) > 8.5). SAR and pharmacokinetic profiling of this series of (3R,6R,7R)-2,5 diketopiperazines together with the introduction of an ortho F group in the 7 aryl ring to improve rat pK has culminated in the 2',4' difluorophenyldiketopiperazine derivative 37, a highly potent oxytocin antagonist against the human oxytocin receptor (pK(i) = 8.9) that has >1000-fold selectivity over all three vasopressin receptors V1a, V2, and V1b. It has good bioavailability (46%) in the rat and moderate bioavailability (13-31%) in the dog and is more active in vivo in the rat than atosiban (rat DR(10) = 0.44 mg/kg iv). PMID- 16250657 TI - Structure-based virtual screening for low molecular weight chemical starting points for dipeptidyl peptidase IV inhibitors. AB - Structure-based virtual screening was performed against the target dipeptidyl peptidase IV (DPP-IV) to identify good chemical starting points for medicinal chemistry. A database of available compounds was filtered by calculated physical properties and undesired chemistry. This database was matched against two in house designed DPP-IV pharmacophores, and the hits from these pharmacophore searches were docked into a DPP-IV crystal structure. Compounds were then selected for testing and 51 active compounds were identified from a list of 4000 compounds tested. These had activities ranging from 30% to 82% when tested at a concentration of 30 microM in an enzyme inhibition assay. PMID- 16250658 TI - Extraction and visualization of potential pharmacophore points using support vector machines: application to ligand-based virtual screening for COX-2 inhibitors. AB - Support vector machines (SVM) were trained to predict cyclooxygenase 2 (COX-2) and thrombin inhibitors. The classifiers were obtained using sets of known COX-2 and thrombin inhibitors as "positive examples" and a large collection of screening compounds as "negative examples". Molecules were encoded by topological pharmacophore-point triangles. In retrospective virtual screening, 50-90% of the known active compounds were listed within the first 0.1% of the ranked database. To check the validity of the constructed classifiers, we developed a method for feature extraction and visualization using SVM. As a result, potential pharmacophore points were weighted according to their importance for COX-2 and thrombin inhibition. Known thrombin and COX-2 pharmacophore points were correctly recognized by the machine learning system. In a prospective virtual screening study, several potential COX-2 inhibitors were predicted and tested in a cellular activity assay. A benzimidazole derivative exhibited significant inhibitory activity with an IC(50) of 0.2 microM, which is better than Celecoxib in our assay. It was demonstrated that the SVM machine-learning method can be used in virtual screening and be analyzed in a human-interpretable way that results in a set of rules for designing novel molecules. PMID- 16250659 TI - Total synthesis of two novel subpicomolar sarco/endoplasmatic reticulum Ca2+ ATPase inhibitors designed by an analysis of the binding site of thapsigargin. AB - Analysis of molecular interaction fields based on the published crystal structure of thapsigargin bound to the sarco/endoplasmatic reticulum Ca(2+)-ATPase and analysis of the volume and shape of the ligand binding site and of the SERCA thapsigargin interactions have enabled design of two new compounds inhibiting SERCA in the subpicomolar range. The two inhibitors were synthesized using (S) carvone as starting material and found to be 3 and 10 times more potent than thapsigargin. PMID- 16250660 TI - Design and studies of novel 5-substituted alkynylpyrimidine nucleosides as potent inhibitors of mycobacteria. AB - We herein report a new category of 5-substituted pyrimidine nucleosides as potent inhibitors of mycobacteria. A series of 5-alkynyl derivatives of 2'-deoxyuridine (1-8), 2'-deoxycytidine (9-14), uridine (15-17), and 2'-O-methyluridine (18, 19) were synthesized and evaluated for their antimycobacterial activity in vitro. 5 Decynyl, 5-dodecynyl, and 5-tetradecynyl derivatives showed the highest antimycobacterial potency against M. bovis and M. avium, with the 2'-deoxyribose derivatives being more effective than the ribose analogues. Nucleosides bearing short alkynyl side chains 5-ethynyl, 5-propynyl, 5-pentynyl, and 5-heptynyl were mostly not inhibitory. Incorporation of a phenylethynyl function at the 5 position diminished the antimicrobial effect. Furthermore, related bicyclic analogues (20-24) were devoid of antimycobacterial activity, indicating that an acyclic side chain at the C-5 position of the pyrimidine ring is essential for potent activity. Compounds 1-17 were synthesized by the Pd-catalyzed coupling reactions of respective alkynes with 5-iodo derivatives of 2'-deoxyuridine, 2' deoxycytidine, and uridine. Intramolecular cyclization of 1 and 3-6 in the presence of Cu afforded the corresponding bicyclic compounds 20-24. The investigated nucleosides are recognized here for the first time to be potent inhibitors of mycobacteria. This class of compounds could be of interest for lead optimization as antimycobacterial agents. PMID- 16250661 TI - 11C-labeling of n-[4-[4-(2,3-dichlorophenyl)piperazin-1-yl]butyl]arylcarboxamide derivatives and evaluation as potential radioligands for PET imaging of dopamine D3 receptors. AB - The selective dopamine D(3) receptor ligands N-4-[4-[(2,3 dichlorophenyl)piperazin-1-yl]butyl]1-methoxy-2-naphthalencarboxamide (1) and N-4 [4-[(2,3-dichlorophenyl)piperazin-1-yl]butyl]-7-methoxy-2-benzofurancarboxamide (2) were labeled with (11)C (t(1/2) = 20.4 min) as potential radioligands for the noninvasive assessment of the dopamine D(3) neurotransmission system in vivo with positron emission tomography (PET). The radiosynthesis consisted in an O methylation of the des-methyl precursors N-[4-[4-(2,3-dichlorophenyl)piperazin-1 yl]butyl]-1-hydroxy-2-naphthalenecarboxamide (3) and N-[4-[4-(2,3 dichlorophenyl)piperazin-1-yl]butyl]-7-hydroxy-2-benzofurancarboxamide (4) with [(11)C]methyl iodide using tBuOK/HMPA and KOH/DMSO, respectively. The radiotracers [(11)C]1 and [(11)C]2 were obtained in 35 min with over 99% radiochemical purity, 74 +/- 37 GBq/mumol of specific radioactivity, 13% and 26% radiochemical yield (EOB, decay-corrected). Distribution studies in rats demonstrated that the new tracers [(11)C]1 and [(11)C]2 cross the blood-brain barrier and localize in the brain. However, the kinetics of cerebral uptake did not reflect the regional expression of the D(3) receptors. Despite their in vitro pharmacological profile, [(11)C]1 and [(11)C]2 do not display an in vivo behavior suitable to image D(3) receptor expression using PET. PMID- 16250662 TI - Synthesis of 5-nitro-2-furancarbohydrazides and their cis diamminedichloroplatinum complexes as bitopic and irreversible human thioredoxin reductase inhibitors. AB - The human selenoprotein thioredoxin reductase is involved in antioxidant defense and DNA synthesis. As increased thioredoxin reductase levels are associated with drug sensitivity to cisplatin and drug resistance in tumor cells, this enzyme represents a promising target for the development of cytostatic agents. To optimize the potential of the widely used cisplatin to inhibit the human thioredoxin reductase and therefore to overcome cisplatin resistance, we developed and synthesized four cis-diamminedichloroplatinum complexes of the lead 5-nitro-2-furancarbohydrazide 8 selected from high-throughput screening. Detailed kinetics revealed that the isolated fragments, 5-nitro-2-furancarbohydrazide and cisplatin itself, bind with micromolar affinities at two different subsites of the human enzyme. By tethering both fragments four nitrofuran-based cis diamminedichloroplatinum complexes 13a-c and 20 were synthesized and identified as bi-ligand irreversible inhibitors of the human enzyme with nanomolar affinities. Studies with mutant enzymes clearly demonstrate the penultimate selenocysteine residue as the prime target of the synthesized cis diamminedichloroplatinum complexes. PMID- 16250664 TI - Enhancing the effectiveness of similarity-based virtual screening using nearest neighbor information. AB - We test the hypothesis that fusing the outputs of similarity searches based on a single bioactive reference structure and on its nearest neighbors (of unknown activity) is more effective (in terms of numbers of high-ranked active structures) than a similarity search involving just the reference structure. This turbo similarity searching approach provides a simple way to enhance the effectiveness of simulated virtual screening searches of the MDL Drug Data Report database. PMID- 16250663 TI - Synthesis and structure-activity relationships of suramin-derived P2Y11 receptor antagonists with nanomolar potency. AB - Selective and potent P2Y(11) receptor antagonists have yet to be developed, thus impeding an evaluation of this G protein-coupled receptor mainly expressed on immune cells. Taking suramin with moderate inhibitory potency as a template, 18 ureas with variations of the methyl groups of suramin and their precursors were functionally tested at P2Y(11), P2Y(1), and P2Y(2) receptors. Fluorine substitution of the methyl groups of suramin led to the first nanomolar P2Y(11) antagonist (8f, NF157, pK(i): 7.35). For selectivity, 8f was also tested at various P2X receptors. 8f displayed selectivity for P2Y(11) over P2Y(1) (>650 fold), P2Y(2) (>650-fold), P2X(2) (3-fold), P2X(3) (8-fold), P2X(4) (>22-fold), and P2X(7) (>67-fold) but no selectivity over P2X(1). QSAR studies confirm that residues with favored resonance and size parameters in the aromatic linker region can indeed lead to an increased potency as is the case for 8f. A symmetric structure linking two anionic clusters seems to be required for bioactivity. 8f may be helpful for studies evaluating the physiological role of P2Y(11) receptors. PMID- 16250665 TI - Discovery and biological evaluation of the novel naturally occurring diterpene pepluanone as antiinflammatory agent. AB - From the whole plant of Euphorbia peplus L., a new diterpene based on a rare pepluane skeleton, named pepluanone (1), was isolated together with a known pepluane diterpene (2). The stereostructure of pepluanone was determined on the basis of an extensive NMR study, MS data, and chemical reaction. The ability of these compounds to act as antiinflammatory agents has been evaluated for the first time by in vivo tests on carrageenin-induced rat paw edema, an experimental model of acute inflammation. Comparison of the bioactivity of pepluanone and compound 2 in terms of chemical structure, evidenced the high efficiency of pepluanone and the absence of appreciable activity for compound 2, thus giving a first insight into the structure-activity relationship. Further in vitro experiments performed on pepluanone let us hypothesize that its activity could be explained in reducing the production of nitric oxide, prostaglandin E(2), and TNF alpha by inhibiting the expression of inducible nitric oxide synthase, cyclooxygenase-2, and TNF-alpha mRNA through the down-regulation of NF-kappaB binding activity. PMID- 16250666 TI - Synthesis and antibacterial activity of 3-substituted-6-(3-ethyl-4 methylanilino)uracils. AB - Numerous 3-substituted-6-(3-ethyl-4-methylanilino)uracils (EMAU) have been synthesized and screened for their capacity to inhibit the replication-specific bacterial DNA polymerase IIIC (pol IIIC) and the growth of Gram+ bacteria in culture. Direct alkylation of 2-methoxy-6-amino-4-pyrimidone produced the N3 substituted derivatives, which were separated from the byproduct 4-alkoxy analogues. The N3-substituted derivatives were heated with a mixture of 3-ethyl-4 methylaniline and its hydrochloride to effect displacement of the 6-amino group and simultaneous demethylation of the 2-methoxy group to yield target compounds in good yields. Certain intermediates, e.g. the 3-(iodoalkyl) compounds, were converted to a variety of (3-substituted-alkyl)-EMAUs by displacement. Most compounds were potent competitive inhibitors of pol IIIC (K(i)s 0.02-0.5 microM), and those with neutral, moderately polar 3-substituents had potent antibacterial activity against Gram+ organisms in culture (MICs 0.125-10 microg/mL). Several compounds protected mice from lethal intraperitoneal (ip) infections with S. aureus (Smith) when given by the ip route. A water soluble derivative, 3-(4 morpholinylbutyl)-EMAU hydrochloride, given subcutaneously, prolonged the life of infected mice in a dose dependent manner. PMID- 16250667 TI - Orally active benzoxazole derivative as 5-HT3 receptor partial agonist for treatment of diarrhea-predominant irritable bowel syndrome. AB - During our search for therapeutic agents to treat diarrhea-predominant IBS, we found that 2-substituted benzoxazole derivatives have a characteristic 5-HT(3) receptor partial agonist activity with high affinity. Some of these compounds showed high in vitro metabolical stability, and 6g showed marked antidiarrhetic activity with little side effect of constipation in in vivo tests. Our results indicate that 5-HT(3) receptor partial agonists might be superior as therapeutic agents to the drugs currently used for IBS treatment. PMID- 16250668 TI - Synthesis, radiosynthesis, and biological evaluation of carbon-11 and fluorine-18 (N-fluoroalkyl) labeled 2beta-carbomethoxy-3beta-(4'-(3-furyl)phenyl)tropanes and -nortropanes: candidate radioligands for in vivo imaging of the serotonin transporter with positron emission tomography. AB - 2beta-Carbomethoxy-3beta-(4'-(3-furyl)phenyl)nortropane (1) was synthesized along with the N-methyl (2), N-fluoroethyl (3), N-fluoropropyl (4), and N-fluorobutyl (5) derivatives. The binding affinity for each compound to the human serotonin, dopamine, and norepinephrine transporters was determined using transfected HEK 293 cells. Radiolabeling and microPET brain imaging studies were performed with [(11)C]1, [(11)C]2, and [(18)F]3 to determine their utility as in vivo imaging agents. PMID- 16250669 TI - Pharmacophore-based design of HIV-1 integrase strand-transfer inhibitors. AB - Using a training set of diketo-like acid HIV-1 integrase (IN) strand-transfer inhibitors, a 3D pharmacophore model was derived having quantitative predictive ability in terms of activity. The best statistical hypothesis consisted of four features (one hydrophobic aromatic region, two hydrogen-bond acceptors, and one hydrogen-bond donor) with r of 0.96. The resulting pharmacophore model guided the rational design of benzylindoles as new potent IN inhibitors, whose microwave assisted synthesis and biological evaluation are reported. PMID- 16250670 TI - Pharmacological and rAAV gene therapy rescue of visual functions in a blind mouse model of Leber congenital amaurosis. AB - BACKGROUND: Leber congenital amaurosis (LCA), a heterogeneous early-onset retinal dystrophy, accounts for approximately 15% of inherited congenital blindness. One cause of LCA is loss of the enzyme lecithin:retinol acyl transferase (LRAT), which is required for regeneration of the visual photopigment in the retina. METHODS AND FINDINGS: An animal model of LCA, the Lrat-/- mouse, recapitulates clinical features of the human disease. Here, we report that two interventions- intraocular gene therapy and oral pharmacologic treatment with novel retinoid compounds--each restore retinal function to Lrat-/- mice. Gene therapy using intraocular injection of recombinant adeno-associated virus carrying the Lrat gene successfully restored electroretinographic responses to approximately 50% of wild-type levels (p < 0.05 versus wild-type and knockout controls), and pupillary light responses (PLRs) of Lrat-/- mice increased approximately 2.5 log units (p < 0.05). Pharmacological intervention with orally administered pro-drugs 9-cis retinyl acetate and 9-cis-retinyl succinate (which chemically bypass the LRAT catalyzed step in chromophore regeneration) also caused long-lasting restoration of retinal function in LRAT-deficient mice and increased ERG response from approximately 5% of wild-type levels in Lrat-/- mice to approximately 50% of wild type levels in treated Lrat-/- mice (p < 0.05 versus wild-type and knockout controls). The interventions produced markedly increased levels of visual pigment from undetectable levels to 600 pmoles per eye in retinoid treated mice, and approximately 1,000-fold improvements in PLR and electroretinogram sensitivity. The techniques were complementary when combined. CONCLUSION: Intraocular gene therapy and pharmacologic bypass provide highly effective and complementary means for restoring retinal function in this animal model of human hereditary blindness. These complementary methods offer hope of developing treatment to restore vision in humans with certain forms of hereditary congenital blindness. PMID- 16250673 TI - Assessing audiological, pathophysiological, and psychological variables in chronic tinnitus: a study of reliability and search for prognostic factors. AB - The development and course of chronic tinnitus are determined by both biological and psychological factors. To combine these different sources of data, we developed a standardized interview to assess tinnitus history, summarize audiological findings, screen for etiological conditions, and explore tinnitus related psychological complaints (Structured Tinnitus Interview). The results of a test-retest study with 65 tinnitus inpatients show that most of these components can be assessed with acceptable or high reliability. Further data based on 166 patients demonstrate that tinnitus annoyance was to some extent different from patterns of general psychological complaints, although there were medium intercorrelations with depression. Significant predictors of tinnitus annoyance were (a) continuous tinnitus without intervals, (b) hearing loss, (c) increasing tinnitus loudness over time, (d) poor maskability, (e) history of sudden hearing loss, and (f) associated craniomandibular disorder. Psychological distress was not significantly increased in patients whose tinnitus was associated to vascular disorder, cervical spine dysfunction, acoustic trauma, Menihre's disease, or neurological disorder. PMID- 16250671 TI - Paclitaxel modulates TGFbeta signaling in scleroderma skin grafts in immunodeficient mice. AB - BACKGROUND: Systemic sclerosis (SSc) is characterized by excessive fibrosis and obliterative vascular lesions. Abnormal TGFbeta activation is implicated in the pathogenesis of SSc. Aberrant TGFbeta/Smad signaling can be controlled by stabilization of microtubules with paclitaxel. METHODS AND FINDINGS: SSc and healthy human skin biopsies were incubated in the presence or absence of paclitaxel followed by transplantation into severe combined immunodeficient mice. TGFbeta signaling, fibrosis, and neovessel formation were evaluated by quantitative RT-PCR and immunohistochemical staining. Paclitaxel markedly suppressed Smad2 and Smad3 phosphorylation and collagen deposition in SSc grafts. As a result, the autonomous maintenance/reconstitution of the SSc phenotype was prevented. Remarkably, SSc grafts showed a 2-fold increase in neovessel formation relative to normal grafts, regardless of paclitaxel treatment. Angiogenesis in SSc grafts was associated with a substantial increase in mouse PECAM-1 expression, indicating the mouse origin of the neovascular cells. CONCLUSION: Low dose paclitaxel can significantly suppress TGFbeta/Smad activity and lessen fibrosis in SCID mice. Transplantation of SSc skin into SCID mice elicits a strong angiogenesis-an effect not affected by paclitaxel. Although prolonged chemotherapy with paclitaxel at higher doses is associated with pro-fibrotic and anti-angiogenic changes, the findings described here indicate that low-dose paclitaxel may have therapeutic benefits for SSc via modulating TGFbeta signaling. PMID- 16250672 TI - Substance use and its relationship to depression, anxiety, and isolation among youth living with HIV. AB - Reductions from lifetime to recent levels of substance use, the time since HIV diagnosis, physical health symptoms, CD4 counts, emotional distress, and social supports were examined among 227 (20% female; 22% African American, 27% Anglo, 35% Latino) youth living with HIV (YLH) ages 13 to 24 years. Substance use pervaded the lives of these youth. Male YLH had used more drugs., more often, and for longer periods than women. However, there had been major reductions in use. Being male, having high emotional distress, and having fewer negative social supports were significantly associated with greater reductions in substance use. There was a trend for the length of time that an individual was seropositive being associated with reductions in substance use. The counterintuitive findings suggest that there must be a re-examination of the role of the social support networks of youth living with HIV, as well as the ways in which emotional distress interacts with risk behaviors. PMID- 16250674 TI - Incidence of cardiac events in hypertensive men related to adaptive behavior in stressful encounters. AB - The Serial Color Word Test was administered at baseline to 253 hypertensive men participating in the prospective cohort study of cardiovascular diseases "Men born in 1914" in Malmo, Sweden. This test of psychological adaptation to a stressful encounter was used to investigate whether susceptibility to stress moderates the risk of a cardiac event in association with hypertension. Adaptive behavior, as measured by test performance, can be categorized in two dimensions. The regression dimension refers to linear change of time spent in the test session whereas the variability dimension refers to nonlinear change. Both dimensions consist of four different patterns. At follow-up (mean time = 8.2 +/- 3.5 years), the risk of a cardiac event varied between men with different adaptive patterns. One pattern, the Cumulative-Dissociative pattern of the variability dimension, characterized by a discontinuous and fluctuating time consumption, was associated to an almost three-fold risk of a cardiac event during follow-up (relative risk [RR], 2.99; 95% confidence interval [CI], 1.33 - 6.70, p = .010) after adjustment for medical-, socioeconomic-, and lifestyle related factors. No association existed between adaptive patterns and overall mortality. PMID- 16250675 TI - Overcommitment predicts restenosis after coronary angioplasty in cardiac patients. AB - The objective of this study is to examine the role of a particular stress enhancing psychosocial risk factor, termed overcommitment, in predicting restenosis after successful percutaneous transluminal coronary angioplasty (PTCA). Overcommitment defines a personal pattern of coping with demands characterized by excessive striving in combination with a strong desire of being approved and esteemed. One hundred six consecutive male patients with coronary artery disease who underwent PTCA were followed over a mean of 12 months. The restenosis rate as defined by quantitative angiography was 34%. Multivariate analysis revealed independent effects of high density lipoprotein cholesterol (odds ratio [OR] 3.19), age (OR 3.43), and overcommitment (OR 2.86) on risk of restenosis. In conclusion, a stress-enhancing psychosocial person characteristic termed overcommitment acts as an independent predictor of coronary restenosis after PTCA. As overcommitment is subject to cognitive-behavioral intervention, results have implications for a more comprehensive approach to secondary prevention in cardiac patients. PMID- 16250676 TI - Cortisol responses to marital conflict depend on marital interaction quality. AB - A sample of 80 couples were videotaped discussing a marital conflict and were then grouped in 3 types according to their interaction behavior: both partners displaying predominantly negative behavior ( N = 36 couples); both partners showing positive behavior ( N = 26); and couples showing asymmetric behavior (one positive, one negative; N = 16). Positive o r negative in this context refers to the empirically defined quality of speaker and listener skills by the Kategoriensystem Partnerschaftlicher Interaktion. Psychophysiological responses were measured 5 times, both before and after the conflict discussion. Participants rated their overall marital quality, the number of marital problems, and their actual cognitions and emotions. Overall responses to the conflict revealed a greater cortisol response in women than in men. Couples grouped according to their interaction style showed significant differences in cortisol responses: In couples with positive interaction, cortisol increased markedly, whereas couples with negative interaction showed a nonresponse, that is, a slight decrease expected with diurnal variation, although they rated the actual conflict discussion as "stressful" like the other groups of couples. We conclude that marital interaction directly affects physiological responses to a conflict depending on interaction quality. PMID- 16250677 TI - Mammography adherence and beliefs in a sample of low-income African American women. AB - The purpose of this article is to describe the relation of perceptions of perceived breast cancer risks and perceived benefits and barriers to mammography and stage of mammography adherence in a convenience sample of low-income African American women. The theoretical framework of the Health Belief Model and the Transtheoretical Model were used to identify concepts and stage of mammography adherence. Data were obtained in waiting rooms of multipurpose centers. Scores for susceptibility and benefits were lowest for those who were in (a) precontemplation (had not thought about having a mammogram); as compared to (b) contemplation (had thought about having a mammogram, but not yet acted); (c) action (had a mammogram as recommended by the American Cancer Society); and (d) relapse (had a mammogram in the past, but overdue). Barriers scores were highest for those who had not had a mammogram (precontemplators and contemplators). In addition, individual barriers were significantly lower for women in action. Results have implications for interventions to increase screening in low-income African American women. PMID- 16250678 TI - Morbidity and mortality following bone marrow transplantation: predictive utility of pre-BMT affective functioning, compliance, and social support stability. AB - This study investigated the utility of 3 pretransplant psychological variables (affective functioning, compliance, social support stability) in predicting subsequent bone marrow transplantation (BMT) health outcomes. The pre-BMT psychological evaluations of 92 BMT recipients were coded along the specified psychological dimensions and used to predict post-BMT survival status and health related quality of life. Data analyses showed that, in addition to medical risk status (low) and quality of the marrow graft (histocompatible), higher levels of pre-BMT affective functioning and social support stability significantly predicted survival status (i.e., alive) and higher levels of quality of life. These findings have important implications for the role of psychological assessment prior to BMT and the need for interventions designed to enhance psychological functioning and subsequent health outcomes following BMT. PMID- 16250679 TI - The initial development of an instrument for the description of "otitis media with effusion specific behavior" in young children. AB - The objective of this study was a preliminary evaluation of a newly developed questionnaire concerning behavior of young children with recurrent Otitis Media with Effusion (OME) and documenting behavioral change after tympanostomy tube insertion. A sample of 95 parent-child pairs were interviewed using an age specific questionnaire: after diagnosing OME and scheduling for surgery, at the day care theatre a few weeks later, and 6 weeks after the intervention. The levels of internal consistency were varying from still insufficient to fairly good for the different subscales of the construct "OME-specific behavior" and these levels were comparable for both age specific questionnaires. As expected, parental report proved to be stable during time, but behavioral changes were reported after tympanostomy tubes insertion. The questionnaires seem to assess a decline in behavior expected to be specific for OME. Implications for future research are discussed, while considering the influence of parental concerns on behavioral reports. PMID- 16250680 TI - Increasing constructive anger verbal behavior decreases resting blood pressure: a secondary analysis of a randomized controlled hostility intervention. AB - We hypothesized that increasing anger verbal behavior in an assertive, constructively motivated style should decrease resting blood pressure (BP) and that this behavior may be one mechanism through which hostility relates to BP. We tested this hypothesis by conducting secondary analyses on a single-blind, matched, randomized controlled study of hostility modification and BP. A total of 22 high-hostile male patients with coronary heart disease were matched on age and hostility level and were randomly assigned to either an 8-week cognitive behavioral hostility treatment (n = 10) or an information-control group (n = 12). Patients were reassessed after treatment and at 2-month follow-up on hostility, observed anger expression, and resting BP. We found that decreases in hostility predicted increases in constructive anger behavior-verbal component, which in turn predicted decreases in resting BP at follow-up. Thus, one of the mechanisms underlying the hostility-BP association may be the lack of constructive anger expression. PMID- 16250681 TI - Vital exhaustion as a risk factor of myocardial infarction: a case-control study in Venezuela. AB - In this study, 32 first myocardial infarction (MI) cases and 42 healthy controls were compared with respect to vital exhaustion (VE), a state characterized by loss of energy, increased irritability, and feelings of demoralization. This state has been found to precede the onset of cardiac events. Participants also responded to questionnaires on Type A behavior, anger expression (Anger In, Anger Out, and Anger Control), and positive and negative self-concept. Results showed that VE discriminated well between MI patients and controls (Odds Ratio [OR] = 15.42, 95% confidence interval = 3.92-60.67) even when controlling for age, smoking, and exercise. The odds ratio decreased to 12.34 when controlling for socioeconomic status. Groups also differed in Anger In but not in Anger Control, Anger Out, negative or positive self-concept. Anger In was correlated to VE in all participants pointing to the relevance of withholding emotions in relation to exhaustion. Exhaustion was strongly associated with negative self-concept in the MI cases group only but significantly discriminated between cases and controls when adjusted for negative self-concept. Summarizing, the results show that, as has previously been found in other countries, in Venezuela VE is a precursor of MI. PMID- 16250682 TI - Everyday stress and Crohn's disease activity: a time series analysis of 20 single cases. AB - In a 84-day long-term study involving 20 patients with active Crohn's disease, the relation between daily stress and symptomatology was investigated. The diagnoses were verified clinically, endoscopically, radiologically, and histologically. The Crohn symptomatology was assessed by a daily symptoms diary. The degree of everyday stress was measured with a daily hassles scale. Additionally, the disease activity was measured 4 times. The data were evaluated using a time series analysis method following the ARIMA model (autoregressive integrative moving average model), which takes the autocorrelation of the data into account. In 55% of the cases, a significant correlation between daily hassles and disease activity was found. Patients whose symptomatology responded to stress did not differ from nonresponders in age, duration of the disease, disease activity, frequency of abdominal surgery, partnership situation, or statements concerning their average level of daily hassles. Disease activity in general did not change during the observation period. A verified influence of daily hassles on Crohn symptomatology as a rule occurs on the same day with time lag 0. According to these results, the group of Crohn's disease patients is not homogenous. The widely held assumption that daily hassles influence the symptomatology cannot generally be confirmed. Variations may result from different ways of coping with stress. Long-term effects of daily hassles on the disease are not clear. PMID- 16250684 TI - Reasons for drinking in relation to problem drinking behavior in a sample of Japanese high school students. AB - To clarify reasons for drinking in relation to problem drinking behavior, 494 male students, aged 15 to 18 years old and attending high schools in Tokyo, Japan, were examined by self-rating questionnaires including the Kuriharna Alcoholism Screening Test (KAST). Three hundred and forty-two students (69%) completed the questionnaires, of whom 143 indicated that they were current drinkers of alcohol (42% of respondents). Of the 143,16 (11%) reported, through the KAST, experience of problem drinking behavior. A factor analysis of 36 reasons for drinking identified 5 factors: Escapism, Sociability, Tension Reduction, Acting Like a Man, and Enjoyment/Home. A logistic regression analysis showed that drinking because of Escapism, Sociability, and Tension Reduction was significantly related to problem drinking behavior. Also, there was significant correlation between the 3 factors and the number of KAST items experienced by the 143 respondents. Our findings suggest that the factors of Escapism, Sociability, and Tension Reduction lead to problem drinking in Japanese high school students. PMID- 16250685 TI - The effect of marital status on survival in late-stage cancer patients: an analysis based on surveillance, epidemiology, and end results (SEER) data, in the United States. AB - Recently findings that marital status is associated with survival in patients with late-stage prostate cancer led to an examination of the generalizability of this association for all cancers. We restricted the investigation to patients with late-stage cancer using population-based data collected from 261,070 patients with late-stage cancer at multiple sites in the United States to determine relations between marital status and survival. After controlling for age, race, and treatment, married patients with cancers of all major primary sites had significantly better survival than single, separated, divorced, or widowed patients. Although single and widowed patients had the poorest prognosis in general, single patients appeared to show the most consistently poor survival across the different types of cancers. Survival differences by marital status were more pronounced in men than in women. This observation raises the possibility that some characteristics associated with being married delay death from cancer. These findings require investigators to ask new questions about the effect of being married and its possible correlates, such as general health status, access to health care, and socioeconomic status. Known correlates of marital status, such as available social support and social isolation also merit attention in relation to these findings. PMID- 16250683 TI - Current perspectives on symptom perception in asthma: a biomedical and psychological review. AB - Symptom perception in patients with asthma is often inadequate. Patients may fail to perceive serious airway obstruction or suffer from breathlessness without objective cause. These extremes are associated with fatal asthma and excessive use of medicines, respectively. This article covers symptom perception in a multidisciplinary perspective. A presentation of current definitions and methods for studying symptom perception in asthma is followed by a summary of theories on the origin of breathlessness. Next, biomedical and psychological factors influencing symptom perception are examined. Preliminary biomedical research emphasizes neural pathway impairment, but causal factors remain inconclusive, particularly regarding the overperception of symptoms. Psychological studies suggest that the accuracy of symptom perception is influenced by (a) competition between asthmatic and nonasthmatic sensory information, (b) negative emotions, and (c) acquired response tendencies (e.g. habituation to symptoms, repression of symptoms, selective perception, and false interpretation of symptoms). These factors may favor either blunted perception or overperception. Empirical data in support of psychological factors are still insufficient. Methodological problems and procedures to improve symptom perception are discussed. PMID- 16250686 TI - Elevated blood pressure and self-reported symptom complaints, daily hassles, and defensiveness. AB - The association between elevated blood pressure and low rates of self-reported problems has been hypothesized to be mediated by defensiveness. In a population screening study in which 1,120 women and 903 men between 20 and 55 years of age participated, multiple resting home blood pressure measurements were performed and questionnaires were administered measuring symptom complaints, daily hassles, and defensiveness. In women, after control for potential confounders, a low number of self-reported symptoms was associated with elevated blood pressure. However, this effect was not mediated by defensiveness, although repressive defensiveness predicted independently elevated blood pressure in women. In men, no significant associations were obtained. Furthermore, no relations emerged between daily hassles and elevated blood pressure. In conclusion, although defensiveness was more prevalent among women with elevated blood pressure, it does not provide a good explanation for the low rates of self-reported symptoms found in these women. PMID- 16250687 TI - Hostility-related differences in the associations between stress-induced physiological reactivity and lipid concentrations in young healthy women. AB - We examined the relations of fasting lipid levels to stress-induced neuroendocrine and cardiovascular responses as a function of hostility in 36 healthy young women. Participants were women who scored above 17 (n = 23) or below 12 (n = 13) on the Cook-Medley Hostility (Ho) Inventory. Lipids were determined following an overnight fast. Individuals participated in a solvable anagram task, which evoked significant physiological responses in all participants. The Ho group by total serum cholesterol (TSC) and the Ho group by low-density lipoprotein (LDL) cholesterol interactions significantly predicted heart rate (HR) and norepinephrine (Ne) responses. For high Ho women, elevations in TSC and LDL cholesterol were associated with smaller HR responses and larger Ne responses. In contrast, for low Ho women, elevations in TSC and LDL cholesterol were associated with larger HR responses and smaller Ne responses. Results also indicated a significant Ho group by TSC to high density lipoprotein (HDL) cholesterol ratio interaction for cortisol. For high Ho women, larger cortisol responses were associated with a greater TSC to HDL cholesterol ratio. For low Ho women, larger cortisol responses were associated with a smaller TSC to HDL cholesterol ratio. Lastly, higher TSC and LDL cholesterol levels were significantly associated with larger epinephrine responses. The findings suggest that stress-induced physiological responses are differentially associated with fasting lipids as a function of hostility in healthy young women. PMID- 16250688 TI - Behavioral medicine and the management of HIV/AIDS. AB - Psychosocial treatment studies provide a method for conducting causal investigations within a clinical environment. They can also inform about relations between psychosocial or biobehavioral processes on the one hand, and disease on the other. Our studies conducted on HIV-positive (HIV+) homosexual men indicate that a group-based cognitive behavioral stress management (CBSM) intervention can decrease distress, buffer the psychological and immunological sequelae of HIV+ serostatus notification, and improve surveillance of herpes viruses. Decreased dysphoria induced by CBSM appears to be a significant mediator of control over cellular immunity. Poor HIV+ African American women, as well as more affluent gay men, benefit from group-based CBSM, but important gender and sociocultural differences must be taken into account in developing protocols. Adherence to highly active antiretroviral therapy (HAART), coupled with good health behavior, can contain HIV/AIDS in most instances. In contrast, poor HAART adherence coupled with poor health behavior (e.g., unprotected sex) can lead to drug resistance and infection of partners with virulent mutated strains. Thus, now more than ever, behavioral medicine approaches to management and secondary prevention of HIV/AIDS are needed. PMID- 16250689 TI - Development and evaluation of a pilot program to promote exercise among mothers of preschool children. AB - The health benefits of physical activity are well documented, but mothers of young children experience significant barriers to exercise. A 10-week minimal intervention exercise program, involving a weekly meeting at which child care was provided, and guidance in establishing independent exercise was developed on the basis of previous research with this population group. Thirty-two women with children under age 5, and in the contemplation or preparation stages of exercise behavior change, were recruited from Australian play groups. Postprogram measures showed small but significant decreases in body mass index, resting heart rate, and diastolic blood pressure. Three months later, 41% of participants were still active. Family-related constraints were highly disruptive to these participants. This study is limited by its small sample size, lack of controls, and potential biases in recruitment and assessment, but it does suggest that hard-to-reach groups, such as mothers of young children, may be mobilized to exercise if programs explicitly address social contexts and the constraints on individual choice. PMID- 16250690 TI - Clinically relevant cholesterol elevation in anxiety disorders: a comparison with normal controls. AB - Recently, several studies reported elevated cholesterol levels in panic disorder, agoraphobia, and general anxiety disorder, but the clinical relevance is still unsettled. All studies so far have disregarded the possible influence of dietary and physical exercise factors. In this study, 30 patients with different anxiety disorders and 30 normal controls were compared for total cholesterol, low-density lipoprotein (LDL), and cholesterol high-density lipoprotein (HDL) ratio. Dietary and physical exercise habits were measured by self-rating questionnaires. Patients with anxiety disorders had significantly elevated total cholesterol, LDL, and cholesterol/HDL ratios. Patients showed borderline-high or high cholesterol levels almost 3 times as often as control participants. Anxiety specific avoidance of physical exercise and special dietary habits of anxiety patients had a significant but minor impact on differences in cholesterol between both groups. Our data support the assumption that serum cholesterol elevations in anxiety disorder patients are within a clinically relevant range. PMID- 16250692 TI - Examination of the AHA!-Illness relation in male and female university students from Australia, India, and the United States. AB - The search for the link between negative emotions and later health problems continues to be a dominant theme in behavioral medicine. Although researchers have identified significant relations between illness and anger, most studies focus selectively on one anger domain: either the affective, attitudinal, or behavioral component. In this study, we first related all three components of the AHA! (anger, hostility, and aggressive behavior) syndrome to somatic complaint reports in a male and female young adult population drawn from three countries. In subsequent analyses, we related multidimensional characteristics of anger to illness in those young adults who regularly use tobacco and alcohol. In both the total sample and the substance user analyses, we found that the anger-illness profile varied as a function of gender. PMID- 16250691 TI - Family support and cardiovascular responses in married couples during conflict and other interactions. AB - This study examined the relation between perceived support from family and cardiovascular (CV) responses to interaction in 45 married couples, 24 to 50 years old. Gender-specific median splits on Family Support scores from the Brief Social Support Questionnaire defined high versus low support groups. CV responses were obtained from both spouses during baseline and 3 stressors: reading control, conversing about events of the day, and conflict discussion. Men with low support had higher systolic (SBP) and diastolic pressure (DBP) than men with high support or either group of women at baseline and during all task periods (p < .004 and .05). Women with high support did not differ in blood pressure (BP) from women with low support. Men and women with low support had higher vascular resistance indexes (VRI) than those with high support, during all periods (p < .0015), and reported lower dyadic satisfaction and total dyadic adjustment. Because a disproportionate number of non-White participants reported low family support, secondary analyses that included race as a covariate were conducted. Covarying for race did not produce substantial results for any of the measures (SBP, p < .05; DBP, p < .058; VR1, p < .021). Post-hoc analyses were also conducted, in which the marital dyad was considered the unit of analysis. For these analyses, in which both spouses' perceptions of family support as high or low were considered together, the wife's perceived support did not influence the husband's BP, whereas his perceived support did influence his BP (ps < .03). For VRI in both men and women, the spouses' support level, as well as the participant's own support level, affected responses (ps < .02). Thus, high family support is associated with both marital and CV benefits for both husbands and wives, although husbands may benefit more. PMID- 16250693 TI - Aberrant parasympathetic and hemodynamic function distinguishes a subgroup of psychologically distressed individuals with asymptomatic type-I diabetes mellitus. AB - In a previous study, a subgroup of asymptomatic insulin-dependent diabetic individuals (termed IDDM-2) were identified on the basis of diminished parasympathetic cardiac input and elevated heart rate at rest. When compared to another group of asymptomatic IDDM participants (termed IDDM-1), and a nondiabetic healthy control group, the IDDM-2 group displayed elevated blood pressure, supported by elevated total peripheral resistance. Measures of psychological regulation were also taken in this study, and form the basis of this article, which examined whether these IDDM-2 patients differed from the other two groups on these measures. The possible role of glycemic control, IDDM duration, and number of somatic complaints among group differences in psychological regulation was also examined. The IDDM-2 group reported increased psychological distress, as reflected by increased dysphoric or depressive symptoms, trait anxiety, perceived stress, and cynical hostility, as well as decreased optimism and interpersonal, but not family, social support. Glycemic control did not account for any of the group differences in psychological regulation. However, group differences in dysphoria and anxiety were accounted for by differences in somatic complaints, whereas differences in interpersonal social support were accounted for by IDDM duration. Moreover, none of the variables investigated accounted for the diminished optimism of the IDDM-2 group. Therefore, in individuals with IDDM, who would otherwise be considered, after medical examination, as no different from other asymptomatic IDDM individuals, the combination of diminished parasympathetic cardiac input and elevated heart rate was associated with aberrant alterations of both hemodynamic and psychological functioning; the increased psychological distress in these individuals may be influenced, in part, by increased diabetes duration and number of somatic symptoms. PMID- 16250695 TI - Job hassles and cardiovascular reaction patterns among urban bus drivers. AB - Forty-six bus drivers took pan in a longitudinal field study of cardiovascular reactions to urban driving. "Job hassles" were recorded by observers using a standardized list of stress-related events in traffic and on the bus. Measures of blood pressure, heart rate, and ratings of perceived mental strain were obtained after each route segment. Intraindividual correlations between psychophysiological recordings and job hassles were calculated and submitted to cluster analysis. Two reaction patterns were identified, 1 characterized by consistently, although modestly, positive associations between the frequency of job hassles and psychophysiological reactions, the other characterized by a low association between the frequency of hassles and indicators or psychophysiological arousal. The former group displayed significantly higher blood pressure and mental strain ratings in the unwinding phase after work than did the latter group of workers. The results are discussed in terms of rate or "unwinding" after exposure to stressful conditions. PMID- 16250696 TI - Cardiovascular reaction to job stress in middle-aged train drivers. AB - In this study, cardiovascular (CV) response to a standard laboratory challenge was compared to 24-hr noninvasive monitoring of heart rate (HR) and blood pressure (BP) in 30 healthy middle-aged train drivers. Laboratory stress test consisted of the orthostatic test, the cold pressor test, the Valsalva maneuver, the Stroop test, and the numerical square. In addition, the participants completed an extensive questionnaire on their health state and family health history, lifestyle, job stress, social and family support, personality characteristics, and health risk behaviors. In waking activities (leisure time, traveling lo work, preparations for driving, and an uneventful driving) NR and systolic blood pressure (SBP), hut not diastolic blood pressure (DBP), were normal (e.g., mean HR = 78.3, SBP = 128.6, and DBP = 92.3 during driving). In occasional stressful work situations, most participants reacted with a considerable rise in SBP and DBP (maximum values 201 for SBP and 126 for DBP). Interindividual differences in maximum BP reactions to emergency stress were predicted reliably by several psychological characteristics and by the CV reactions to the laboratory psychological challenge. The frequency, intensity, and persistence of psychological and physiological reactions to urgent situations appear to be more relevant measures of the health impact of psychological job stress than are the shift average values of physiological stress markers. PMID- 16250698 TI - Angina pectoris, job strain, and social status: a cross-sectional study of employed urban citizens. PMID- 16250697 TI - Job strain and biological coronary risk factors: a cross-sectional study of male and female workers in a Japanese rural district. AB - To investigate the association between job strain and biological coronary risk factors, a demand-control questionnaire was applied to 138 male (mean age 51.5) and 166 female (50.8) workers in Japanese rural town. In Model I, workers rated as both above the median on demands and below the median on control were defined as a strain group and compared to the rest. In Model II, the effect of a multiplicative term of demands by control was tested once the component main effects were controlled. In both models, possible confounders were controlled. Men in the strain group had higher blood glucose than did the others (Model 1), and the multiplicative term was significantly associated with diastolic blood pressure in men and with Lipoprotein(a) in women (Model II). The results suggest that the demand-control model predicts coronary risk factors in Japanese rural workers, and the associations are different between genders. PMID- 16250699 TI - Who comes to a workplace health risk assessment? AB - Workplace health promotion initiatives have proliferated, but there are difficulties in recruiting employees of lower socioeconomic status and at higher risk of disease. A survey of health behaviors and attitudes was administered in 20 worksites and the opportunity to attend a health risk assessment promoted. Those more likely to attend were women, those of higher occupational prestige, and those from a non-English-speaking background. After adjustment for these variables, the only health behavior associated with attendance was smoking status. Perceived risk of lung cancer was significant, even after adjustment for smoking status. Stage of readiness to change health behaviors was associated with attendance, with those in the preparation stage being more likely to attend than those in the precontemplation stage. However, this association was statistically significant only for fruit and vegetable consumption. There was no relation between attendance and support for health promotion, perceived general health, or other perceived risk of disease. These findings suggest that additional risk communication strategies and environmental support are required to involve those with less prestigious occupations. PMID- 16250700 TI - Durability, dissemination, and institutionalization of worksite tobacco control programs: results from the Working Well trial. AB - Durability, dissemination and institutionalization o f tobacco control activities are reported, based on the Working Well worksite cancer control intervention study (n = 83 worksites). Tobacco control activities increased significantly in intervention worksites as a result of research-supported activities but were not sustained 2 years after the conclusion of the intervention. Intervention sites were more likely than control sites to initiate and maintain structures for institutionalizing programs, such as assigning a committee responsibility for health-promotion programs or providing a budget for health-promoting activities. Dissemination of the program to control worksites had little impact on the level of smoking control activities in control worksites. Although program durability was not a primary aim of this intervention study, these analyses provide an important assessment of program maintenance beyond a funded intervention and underscore the need for additional research to identify effective organizational strategies for institutionalization of worksite health-promotion programs. PMID- 16250702 TI - Acute hemoconcentration during psychological stress: assessment of hemorheologic factors. AB - In this study, we examined the effects of acute psychological stress on hemorheology and hemoconcentration in humans and the associations between stress induced cardiovascular reactivity and hemorheological changes. Stress-induced changes in hemorheology and hemorheological changes were assessed by measuring plasma viscosity, calculated plasma volume, and total plasma protein. Cardiovascular, hemorheologic, and hematologic variables were assessed in 29 healthy men during a 30-min baseline period and a 5-min speech task. Results indicated that the speech task produced a significant increase in plasma viscosity and total plasma protein and a significant decrease in calculated plasma volume. Significant correlations were observed between changes in blood pressure and heart rate and changes in plasma viscosity, total plasma protein, and calculated plasma volume. These results provide direct evidence that acute psychological stress can produce significant changes in hemorheology and hemoconcentration. The most likely mechanism for the stress-induced hemoconcentration effect is a fluid shift from the vascular to the interstitial spaces through increased blood pressure. PMID- 16250701 TI - Pain and stress in sickle cell disease: an analysis of daily pain records. AB - This study examined daily reports of pain, medication use, health care use, and activity reduction in adults with sickle cell disease, and their association with stress. Participants were 53 adults with sickle cell disease. They completed the Daily Hassles questionnaire at the start of the study, and they kept daily records of pain and pain response over the following 14 days. On average, patients reported pain on 6.5 days of the 14-day study period. The average pain intensity rating during a painful episode was 4.4 on a 10-point scale. Pain was most often managed at home. Patients took medication (analgesics and/or narcotics) on 80% of the days they experienced pain, and they were more likely to use medication, particularly narcotics, as pain levels increased. At higher pain levels some patients also utilized a range of health care services. On average, patients also cut back considerably on household and social activities, especially when pain reached a level of over 5 on the 10-point scale. Those who were employed, however, were likely to continue to work, even when in pain. In addition, stress had significant positive associations with average pain intensity as well as reductions in household and social activities. Furthermore, stress predicted activity reductions even after controlling for pain intensity. Stress was unrelated to medication and health care use in this study. PMID- 16250703 TI - Classification of individual differences in cardiovascular responsivity: the contribution of reactor type controlling for race and gender. AB - Classification of 150 normotensive or mildly hypertensive men and women into myocardial, vascular, or mild reactors was accomplished using a regression-based approach. The method was based on the participants' cardiac output (CO) and total peripheral resistance (TPR) reactivity to the speech presentation task. This task purportedly can elicit both myocardial and vascular responses. Cut-scores were based on the y-intercept from the linear regression of the CO reactivity on TPR reactivity and vice versa. A greater percentage of Black men were classified as vascular responders as compared to Black women and White participants. Groups were found to differ on cardiovascular reactivity to the speech preparation, cold pressor, and mirror tracing tasks in predictable ways, after controlling for gender and ethnicity. Groups were also differentiated by ambulatory blood pressure and hypertensive status. The study supports the classification of homogenous groups of participants based on the relative extent to which myocardial or vascular mechanisms dominate the reactivity to stress. PMID- 16250704 TI - Environmental tobacco smoke: association with cardiovascular function at rest and during stress. AB - Chronic exposure to environmental tobacco smoke (ETS) contributes to cardiovascular disease morbidity and mortality, and ETS alters cardiovascular performance during exercise stress. However, no study has examined whether those with ETS exposure have altered cardiovascular functioning during psychological stress, relative to those with no substantial ETS exposure. Seventy-eight healthy, nonsmoking adult men with either high levels of current ETS exposure at home and work or no current or significant lifetime ETS exposure were tested in a stress reactivity protocol. Blood pressure and heart rate were monitored during rest and during two psychological stressors. Those with high ETS exposure had significantly elevated heart rate and blood pressure at baseline, relative to those with no exposure. The groups did not differ on their cardiovascular stress responses. These data suggest that chronic ETS exposure is associated with altered cardiovascular functioning at rest, but not during stress. Results are discussed with regard to the role of ETS on the development of heart disease among nonsmokers. PMID- 16250705 TI - Social cognitive and physical health determinants of exercise adherence for HIV-1 seropositive, early symptomatic men and women. AB - This study examined social cognitive and physical health factors that might explain variations in exercise adherence in a 3-month supervised exercise intervention for a group of mildly symptomatic, HIV-1 seropositive men and women. The social cognitive predictor variables were outcome expectations and self efficacy. The physical health predictor variables included CD4+ cell counts, self report inventories of physical symptoms, and physicians' examinations. Self report inventories of physical symptoms were associated with physicians' examinations and combined into a composite measure of physical health. Criterion variables included exercise adherence rates, VO2max change, and status as a 'remainder' versus 'drop-out.' The composite measure of physical health emerged as a significant predictor of adherence rate and gave perfect prediction of remainers and a moderate prediction of dropouts. No significant associations were observed between the social cognitive predictors and adherence. Results suggest that for this population physical health status is a better predictor of exercise adherence than either perceived self-efficacy or outcome expectancy. PMID- 16250706 TI - Acute stress modulates symptom awareness and hormonal counterregulation during insulin-induced hypoglycemia in healthy individuals. AB - The purpose of this study was to determine whether the perception of hypoglycemia is reduced during acute stress. In Session I each of our 40 healthy male volunteers received a bolus injection of human insulin (0.05 IU/kg) resulting in plasma glucose nadirs of below 2.8 mmo/L. In Session 2 participants received insulin or saline, with half of each group being stressed by having to prepare and give a speech. Data collection at 5- to 25-min intervals included a symptom checklist, blood pressure, heart rate, and blood sampling for measurement of plasma glucose and counterregulatory hormones. Individuals in the stress + insulin group were less sure of having received insulin and ate fewer cookies compared with controls. They reported lower intensity of the hypoglycemic symptoms of palpitations, tremor, dizziness, and blurred vision, in contrast to the reduced subjective and behavioral reactions, they showed the strongest hormonal counterregulation. We conclude that acute stress during hypoglycemia reduces symptom awareness and the ability to detect hypoglycemia. PMID- 16250707 TI - Cardiovascular and perceptual effects of reporting pain during the foot and forehead cold pressor tests. AB - In research involving the cold pressor test, a tacit presumption is often made that reporting pain during stimulation is not in itself reactive. This study examined whether, for the foot and forehead cold pressor tests, activities involved in reporting pain may affect (a) the evoked pattern of cardiovascular response, and (b) the magnitude of self-perceived pain. In 40 normotensive college men, increases in systolic blood pressure were greater during test sessions that included verbal ratings of pain, as compared to sessions in which pain was not reported. In contrast to its effect on physiological activation, reporting pain did not significantly alter the participant's perception of the painfulness of the lest, on recollection shortly after the test. We conclude, therefore, that reporting pain during the cold pressor test may impose significant additional demands on the cardiovascular system, but it does not interfere significantly with the processing of nociceptive information. PMID- 16250708 TI - Increases in salivary lysozyme and IgA concentrations and secretory rates independent of salivary flow rates following viewing of humorous videotape. AB - Several studies have shown increases in salivary, secretory IgA (sIgA) levels following exposure to humor and various relaxation strategies. However, the use of sIgA as a marker of immunocompetence has been questioned. In this study we investigated the potential of an alternative marker, salivary lysozyme (sLys) in addition to sIgA. sIgA and sLys concentrations and secretion rates were measured from saliva samples collected immediately prior to and alter viewing a humorous (situation comedy) and a control videotape (documentary). sLys concentrations and secretion rates were significantly increased after viewing of the humorous videotape (p = .0007 and p = .009, respectively) but not following viewing of the control video. Similarly, increases were obtained for sIgA (p = .01 and p = .05) following viewing of the humorous tape. sIgA concentration decreased following viewing of the control tape (p = .05). Salivary flow rates did not significantly change following the viewing of either the humorous or the control tapes. Changes following the humorous video were significantly different from changes following the control video for both sLys and sIgA levels (concentration, p = .007 and p = .005; secretory rate p = .02 and p = .03), but not for salivary flow rates. The results suggest an enhancement of the innate and specific mucosal immune parameters following exposure to humor, independent of salivary flow. Use of sLys may serve as a useful additional or alternative marker of the effects of humor on mucosal immunity. PMID- 16250709 TI - Construct validation of health-relevant personality traits: interpersonal circumplex and five-factor model analyses of the Aggression Questionnaire. AB - The general literature on personality traits as risk factors for physical illness -as well as the specific literature on health consequences of anger, hostility, and aggressive behavior--often suffers from incomplete or inconsistent construct validation of personality measures. This study illustrates the utility of two conceptual tools in this regard--the five-factor model and the interpersonal circumplex. The similarities and differences among anger, hostility, verbal aggressiveness, and physical aggressiveness as measured by the Buss and Perry (1992) Aggression Questionnaire were identified. Results support the interpretation of anger and hostility as primarily reflecting neurotic hostility and antagonistic hostility to a lesser extent. In contrast, verbal and physical aggressiveness can he seen as primarily reflecting antagonistic hostility, and to a lesser extent neurotic hostility. Further, verbal aggressiveness was associated with hostile dominance, whereas hostility was associated with hostile submissiveness. These findings identify potentially important distinctions among these related constructs and illustrate the potential integrative value of standard validation procedures. PMID- 16250711 TI - Health-related behaviors, social support, and community morale. AB - The relation among health behaviors, social support, and community morale- attitudes toward one's community, such as attachment and social participation- was examined in 597 adults living in a Japanese community. Logistic models revealed that strong spousal support was related to reduced alcohol drinking and enhanced family support was related to a reduction in smoking. The support of friends was positively associated with the frequency of alcohol drinking by men. Among women, family support was positively related to the frequency of consuming Japanese-style food. Community morale was positively associated with consuming Japanese-style side dishes and negatively associated with alcohol consumption. The results indicate that there are varied associations between social relationships and health-related behavior and that these associations are gender related. Knowing the specific associations between an individual's social relationships and his or her health-related behavior will he helpful for community-based intervention. PMID- 16250710 TI - Predicting smoking stage of change among a sample of low socioeconomic status, primary care outpatients: replication and extension using decisional balance and self-efficacy theories. AB - An expanding body of research using the transtheoretical model with both self change and treatment programs suggests that differences in readiness for smoking cessation are important predictors of successful abstinence. Understanding the cognitive processes underlying these differences may hold tremendous potential for improving the efficacy and efficiency of intervention strategies. Decisional balance theory and self-efficacy theory have been used to help explore how and why people move through the stages of change, but they have been validated almost exclusively with middle-class, educated White samples This study sought to investigate whether these theories relate in each other in the same manner among low socioeconomic status (SES) primary care outpatients. Results indicated that variables from decisional balance theory (pros, cons) and self-efficacy theory successfully differentiated stage membership and yielded results consistent with the extant literature. Self-efficacy demonstrated the most powerful association with stage membership, whereas pros, cons, and temptations exhibited varying degrees of association. Clinical implications and special considerations when conducting research and implementing interventions with low-SES smokers are discussed. PMID- 16250712 TI - State anxiety predicts poor psychosocial outcome after coronary bypass surgery. AB - A prospective study was conducted to identify predictors of poor psychosocial adjustment 1 year after coronary bypass graft (CABG) surgery. Fifty-two of 61 consecutive patients (mean age = 66 +/- 9, 3 years, 76% men) were studied before and 1 year after surgery. Psychological, social, and surgical data were assessed. The roles of State and Trait anxiety, emotional reactions, and different coping modes were evaluated. At baseline the patients were divided into 3 groups according to their level of anxiety as assessed by Spielberger's State Anxiety Inventory (STAI). The 3 groups did not differ in any of the basic parameters, except for their subjective experience of symptoms of dyspnoea (p < .01), and quality of life (p < .0001) for the whole patient group 1 year after surgery. However, patients belonging to the moderate-anxiety and high-anxiety groups remained more psychologically distressed (p < .0001) and perceived a higher degree of residual angina pectoris (p < .0001) than did patients who did not display anxiety preoperatively. Dividing the patients into groups according to their answers to the other measures of psychological distress or negative coping strategies yielded similar results. The results indicate that the SATI is a valuable instrument for identifying risk patients for poor adjustment after CABG surgery. PMID- 16250713 TI - Attenuated cortisol response to a standardized stress test in Lithuanian versus Swedish men: the LiVicordia study. AB - Cardiovascular mortality rates of middle-aged men are 4 times higher in Lithuania than in Sweden The difference is not explained by standard risk factors, but our previous findings of pronounced psychosocial stress in Lithuanian men offer a possible explanation. We investigated cortisol and cardiovascular reactivity to a standardized laboratory stress test in population-based random samples of 50-year old men from Vilnius, Lithuania and Linkoping, Sweden. Repeated measures analysis of variance showed that cortisol responses differed between cities (p's < .0001). Mean change of serum cortisol from baseline to 30 min was 18.1 and 88.4 nmol/1 for Vilnius and Linkoping men, respectively (p < .001). In a multivariate analysis, a low peak cortisol response was significantly related to high baseline cortisol, current smoking, and vital exhaustion. The findings suggest a physiological mechanism of chronic psychosocial stress, which may contribute to increased risk for cardiovascular death. PMID- 16250714 TI - Heart rate variability at rest and during mental stress in patients with coronary artery disease: differences in patients with high and low depression scores. AB - This study tested the hypothesis that coronary artery disease (CAD) patients with high depressed mood scores differ in sympatho-vagal balance during mental stress compared to patients with low depressed mood scores. Using electrocardiographic monitoring, heart variability data from spectral analysis and hemodynamic parameters were obtained prior to and during mental stress from 34 men and 7 women. A public speaking task was used as the mental stressor. Patients were grouped by a median split of their Minnesota Multiphasic Personality Inventory Depression score. During mental stress, patients with higher depression scores had greater changes in peak heart rate (p < .05) and low frequency to high frequency power ratio (p < 0.05) than patients with lower scores suggesting a shift toward more sympathetic activity during mental stress. These findings may be related to the reported relation between depression and survival risk in patients with CAD. PMID- 16250715 TI - Effects of parental history of hypertension and urbanization on blood pressure in Zimbabweans. AB - The increasing prevalence of essential hypertension is a growing public health concern for Zimbabwe and other African countries. Two important risk factors for hypertension are urbanization and parental history of hypertension. The relations among parental history of hypertension, urbanization, and blood pressures (BPs) are poorly understood. The objective of this study is to clarify these relations in a population of urbanized, African, young adults. The relation between parental history of hypertension and urbanization on resting BP's and Bp responses to a mental arithmetic stressor was examined in a group of normotensive, Black medical students with (n = 36) and without (n = 34) a parental history of high BP, and with (n = 49) and without (n = 19) a parental history of urbanization. Results indicate that those with a positive parental history counterparts. Further, those with parents residing in urban areas had higher resting SBPs than those with parents residing in rural areas. However, no reactivity differences were apparent between the urban and rural parent groups. These data suggest that although parental history for hypertension influences both resting and reactivity BP's parental history of urbanization may influence only resting BP. PMID- 16250716 TI - Altered cardiovascular/pain regulatory relationships in chronic pain. AB - In healthy individuals, there is an inverse relationship between resting blood pressure (BP) and pain sensitivity. This study examined possible dysregulation of this adaptive relation in chronic pain patients, and tested whether the extent of this dysregulation is a function of pain duration. Continuous resting BP's were assessed for 5 min after a 5-min rest period in 121 chronic benign pain patients. Unlike the inverse relationship observed previously in normals, mean resting diastolic. BPs during the assessment period were correlated positively with ratings of pain severity. A Pain Duration x Systolic BP interaction emerged (p < .05) such that the magnitude of the BP-pain relation was greatest in patients with the longest duration of pain, r(38) = .50, p < .001. A hypothesized progressive alteration in endogenous pain regulatory systems in chronic pain patients was supported. A possible role of endogenous opioid dysfunction in accounting for these alterations is discussed. PMID- 16250719 TI - Prediction of resting cardiovascular functioning in youth with family histories of essential hypertension: a 5-year follow-up. AB - Two hundred forty-six children (96 Whites, of whom 51 were males; 150 African- Americans, of whom 69 were males) with a familial history of essential hypertension (EH) were re-evaluated 5 years after an initial evaluation. During the initial visit anthropometric, demographic, and resting cardiovascular (CV) parameters (designated initial baseline levels) were assessed. These CV parameters (systolic and diastolic blood pressure [BP], heart rate, cardiac output index [CI], and total peripheral resistance index [TPRI] were also measured during postural challenge, a video game challenge, and a cold pressor task. At follow-up, resting CV parameters were again evaluated, and designated as follow-up resting levels. Moderate temporal stability (r range = .43-.56) was observed for all resting CV parameters. Mean stress responses for each CV parameter for all 3 stressors during the initial visit were positively related to the respective CV follow-up resting level. BP stress responses to postural change and video game challenge to be significant independent predictors of future resting BP after controlling for standard EH risk factors. Follow-up resting CI was not predicted by any stress responses, whereas follow-up resting TPRI was predicted by TPRI responses to the video game after controlling for standard Eh risk factors. These results contrast with those from an earlier 1-year follow-up, where stress responses for neither CI nor TPRI predicted follow-up resting levels. It appears that, as children get older, TPRI stress responses play a stronger role in vasoconstrictive function. PMID- 16250717 TI - Effect of satisfaction with social support on blood pressure in normotensive and borderline hypertensive men and women. AB - The relation between blood pressure (BP) and two aspects of social support, perceived satisfaction and structural social support network characteristics, were examined in adults classified as having normal BP or borderline hypertension. Causal BPs were taken by a trained technician on 3 separate visits, each approximately 1 week apart. Participants were categorized as borderline hypertensive if screening systolic blood pressure (SBP) fell in the range 130-160 mmHG and/or diastolic blood pressure (DBP) fell in the range 85-100 mmHg; BPs below these ranges were considered normotensive. Participants underwent ambulatory BP monitoring. Individuals classified as normotensive reported significantly greater support satisfaction than individuals with borderline hypertension. Social network characteristics were not associated with BP. During 24 hr of ambulatory BP monitoring, high perceived support satisfaction was associated with lower SBP and DBP at work, home, and during sleep. These data suggest that perceived satisfaction with social support is associated with lower BP. The presence of social support also may reduce BP levels during daily life. PMID- 16250720 TI - Assessing type a behavior in 8-year-olds: exploring the overlap between the constructs of type a behavior and hyperactivity. AB - This study attempted to examine, by using behavioral observations, the overlap found between Type A behavior, as measured by the Matthews Youth Test for Health (MYTH) questionnaire, and measures of hyperactivity in children. The sample consisted of 91 children (46 boys and 45 girls) between the ages of 8 and 9 who were part of a longitudinal study in Uppsala, Sweden. Behavioral observations of Type A behavior and various questionnaire data were used. The behaviorally observed Type A behavior correlated significantly with MYTH Type A behavior, whereas only the MYTH Type A behavior correlated with hyperactivity measured by questionnaires. The results indicated that behavioral observations make it possible to measure Type A behavior as distinct from hyperactivity, implying that the MYTH is too indiscriminate a measure of Type A behavior. This has implications for future measurement and perhaps conceptualization of Type A behavior in children, and thereby also for studying the developmental aspects of Type A behavior. PMID- 16250721 TI - Behavior of 3-year old children in a prospective randomized trial of reduced saturated fat and cholesterol diet since infancy: the STRIP baby project. AB - Interventions aimed at decreased exposure of children to known atherosclerosis risk factors may have untoward behavioral side effects. We examined how children's behavior or parent's perception of the behavior of the children at 3 years of age was influenced by the intervention in a prospective randomized trial that began in infancy and effectively decreased serum cholesterol concentration. This Special Turku coronary Risk factor Intervention Project for babies (STRIP) began when the infant was 7 months old. Half of 1,062 children received individualized dietary counseling at 1 to 3-month intervals during the first 2 years of age and then half-yearly; the other half had an unrestricted diet. At 3 years of age a standardized questionnaire of the child's behavior was sent to 791 families (76% returned the questionnaire). At the onset of the trial the sociodemographic data of the families and serum lipid values of the intervention and control children were similar. Later, mean serum cholesterol values of the intervention children remained constantly at a level 6% to 10% below the values of the control children. At 3 years of age the parental perceptions of the child's behavior suggested minimal differences between the intervention and control children. The intervention children were slightly less jealous and more active and creative but showed slightly more negative signs of behavior (bed wetting, problems in falling asleep, fears) than the controls. We conclude that long-term, individualized dietary and lifestyle intervention that begins in infancy slightly influences children's behavior or parent's recognition of the behavior of the children at the age of 3 years. PMID- 16250722 TI - Development of the child/adolescent activity log: a comprehensive and feasible measure of leisure-time physical activity. AB - The purpose of this study was to test the utility of a new instrument, the Child/Adolescent Activity Log (CAAL), designed to measure children's physical activities in a school setting efficiently, yet comprehensively. At baseline, 459 participants (mean age 12 + 1.4, 59% White, 34% African American, 7% other) were recruited to participate in a 2-year study focusing on patterns and predictions of physical activity at critical school transition points. The CAAL was administered daily for a 1-week period on 6 occasions to this cohort. From the CAAL, information about the pattern of specific activities chosen, average daily duration of activity, and average daily expenditure per kg body weight was obtained. The validity of the log was supported by its (1) correlation with Caltrac readings, (2) relation in the predicted direction with a single-item measure of typical level of physical activity and fitness indices, and (3) expected changes in exercise patterns over time and by gender. If future studies corroborate the psychometric properties and ease of administration of the CAAL, its utility in community-based studies is promising. PMID- 16250723 TI - Coping with pain and surgery: children's and parents' perspectives. AB - This article examines relations between coping with general and postoperative pain. Children's and parent's coping ratings, and the contribution of temperament and coping to postoperative adjustment. Before and after day surgery, 7-to 12 year-olds (n = 124) rated their coping with pain. Parents rated their child's coping and temperament pain and distress were rated on the day of and 2 days following surgery. Coping with general and postoperative pain were moderately correlated (median r = .48). Except for distraction, all types of coping strategies were used more frequently for general than postoperative pain. Correlations between child and parent coping ratings were moderate (median r = .36). After controlling for emotionality and medications, lower levels of emotion focused avoidance and higher levels of distraction were related to lower pain and distress. After controlling for a priori surgical group (no-low pain; moderate high pain), emotionality, and medications, lower levels of emotion-focused avoidance were related to lower pain and distress but distraction was no longer significant in a number of the regressions. Coping with pain has trait-like qualities but differences in the nature and context of pain create differences in strategy use. Level of pain appears to influence the type of coping strategies used. Interventions should target children's use of distraction and minimize us of emotion-focused avoidance. PMID- 16250724 TI - Cognitive coping skills training in children with sickle cell disease pain. AB - This study was designed to examine whether brief training in cognitive coping skills would enhance pain coping strategies and alter pain perception in children and adolescents with sickle cell disease (SCD). Forty-nine participants with SCD were randomly assigned to either a cognitive coping skills condition or a standard care control condition. At pre- and posttesting, coping strategies and pain sensitivity using laboratory pain stimulation were measured. Results indicated that in comparison to the randomly assigned control condition, brief training in cognitive coping skills resulted in decreased negative thinking and lower pain ratings during low intensity laboratory pain stimulation. PMID- 16250725 TI - A behavioral treatment of young migrainous and nonmigrainous headache patients: prediction of treatment success. AB - We evaluated the outcome of a behavioral treatment package in a clinical setting with a group of young (age: 12-22) headache patients, suffering from migrainous or nonmigrainous. Comparison between the experimental (n = 24) and the waiting list control group (n = 15) showed a treatment effect on headache frequency and on the headache index. Using a 50% reduction in the headache activity as a criterion for clinical improvement, 52% of the participants in the experimental group had improved clinically at the end of the treatment. The treated participants were found to have maintained significant improvement at 1-year follow-up. The treatment effect was significantly high for nonmigrainous headache patients than for migraineurs. The most important background predictor of outcome was duration of headache history: youngsters with a longer headache history profited less by the treatment than youngsters with a shorter headache history. Family predictors of pre-post improvement were maternal rewarding of illness behavior and mother-child relationship Those youngsters who reported more rewarding and/or a more positive mother-child relationship profited less by the treatment than those who reported less rewarding and/or a less positive mother child relationship. We conclude that therapists treating young headache patients should be alert to pain-rewarding patterns in the family. PMID- 16250726 TI - The role of monitoring in determining quality of life following treatment for a bone tumor. AB - Interviews were conducted with 34 young people who had previously been treated for a malignant bone tumor around the knee. These interviews focused on the impact of treatment on activities and perceptions of the risk of recurrence and need for future surgery A coding schema based on a "monitoring-blunting" framework was adopted (Miller, 1995). Quality of life was assessed using a generic and disease-specific measure. Based on interview data, respondents were categorized as negativistic monitors, adaptive monitors, and nonmonitors. There were no differences between groups in terms medical indicators (number of operations). Negativistic monitors reported poorer quality of life compared with the other two groups. There was no increase in nonmonitoring with time since diagnosis as reported in previous work. It is suggested that patients' self ratings of quality of life are related to the way in which they monitor information and this may be independent of clinical function. Clinical implications, especially in terms of how potentially threatening information about late-effects of treatment are given to patients, are discussed. PMID- 16250727 TI - Perceived problem-solving ability, stress, and coping in mothers of children with physical disabilities: potential cognitive influences on adjustment. AB - Examined the contribution of perceived problem-solving ability to the adjustment of mothers of children with a physical disability, in conjunction with appraised disability-related stress and approach and avoidance coping, as a further evaluation of Wallander and Varni's Disability-Stress-Coping model emphasizing cognitive processes. One hundred sixteen mothers of children, ages 2 to 20, with spina bifida or cerebral palsy completed measures of relevant constructs. Results show that the mother's appraisal of disability-related stress is strongly associated with her reported maladjustment. However, perceptions of competence in problem solving are associated with better overall adjustment. A portion of this relation appears mediated by coping styLe, as confidence in one's problem-solving ability increases so does the likelihood of selecting more adaptive coping strategies when faced with disability-related stress. Overall, consideration of perceived problem-solving ability appears to make a valuable contribution to the Disability-Stress-Coping model and to current understanding of the experience of coping with raising a child with a disability. Perceived problem-solving ability can also be linked to clinical intervention, as discussed. PMID- 16250728 TI - Glycemic control, self-care behaviors, and psychosocial factors among insulin treated diabetics: a test of an extended health belief model. AB - The relations of diet adherence (DA) and self-monitoring of blood glucose (SMBG) to metabolic control, as measured with glycosylated hemoglobin A(tc) (GHbA(tc), and correlates of self-care were examines among a type I diabetic sample (n = 423). The Health Belief Model (HBM), supplemented by other factors (locus of control, self-efficacy, health value, and social support), was used as a theoretical model. In multiple regression analyses both DA (p<.01) and SMBG (p<.001). SMBG showed strong associations with self-efficacy in SMBG (p,.001) and net benefits of SMBG (p<.001). The revised models explained 14% and 21% of the variation in DA and SMBG, respectively. The results suggest that although perceived net benefits are important determinants of both SMBG and DA, DA is also related to diabetes support, whereas SMBG is more strongly related to perceived self-efficacy. Thus self-care regimen should be planned individually for diabetic patients. PMID- 16250729 TI - Organ donation: who is not willing to commit? Psychological factors influencing the individual's decision to commit to organ donation after death. AB - One hundred thirty-nine students in Israel answered 3 questionnaires, Attitudes Toward Organ Donation, Trait-State Anxiety, and Fear of Death. A minority (17%) had signed an organ-donor card. This was found related to age, religious affiliation, viewing donation as a good idea for oneself, knowing people with organ donor cards, and higher scores on the Fear of Death Scale. The combination of the personality scales explained 5% of the total variance of the 'donor card' variable. Factor analysis identified 3 factors representing the reasons preventing the commitment to donate, 'Avoidance,' 'Lack of Interest,' and 'General Intention.' Cluster analysis showed that different combinations of the factors created 3 different clusters. The personality variables explained 18% of the variance of belonging to a specific nondonor cluster. It was concluded that failure to possess an organ donor card does not necessarily reflect opposition to donation and that there are different dimensions of ambivalence toward the commitment to donate one's organs after death. PMID- 16250730 TI - A preliminary study of the effects of breast-feeding on maternal health. AB - This study is a preliminary investigation of the effects on breast-feeding and cessation of breast-feeding on maternal health. A self-report questionnaire assessed overall health, mood, stress, and psychological and upper respiratory infection symptoms in 14 current, 36 never, and 49 past breast-feeders. Results indicated that breast-feeding did not incur greater health costs to the mother compared with bottle-feeding and that cessation of breastfeeding was associated with worse mood, more stress, and a greater number of psychological symptoms compared with current breast-feeding. Physiological mechanisms, as opposed to personality factors, are discussed as a basis for these findings. PMID- 16250731 TI - Hostility, testosterone, and vascular reactivity to stress: effects of sex. AB - This study investigated the association of personality with cardiovascular stress reactivity (CVR) in men and women. Also, the degree to which testosterone and estradiol reactivity were related to personality and CVR measures was examined. Twenty-six men and 44 women completed the Cook-Medley Hostility Scale, the Beck Depression Inventory, and the Spielberger Trait Anxiety Inventory before speech, Stroop, and math stress. Testosterone (men) and estradiol (subset of women) were sampled once after an initial rest period and again after the last stressor. Cardiovascular reactivity, including cardiac output and total peripheral resistance (TPR), was assessed during stressors. For men, testosterone increased significantly with stress, and testosterone reactivity to stressors was significantly correlated with hostility. However, stepwise multiple regression revealed that hostility was the only independent predictor of CVR to speech, math, and Stroop stress in men, accounting for 13%-32% of the variance in TPR. Baseline systolic blood pressure explained 22% of the variance in TPR reactivity to speech preparation. No evidence was obtained to suggest that hostility, depressive mood, or anxiety predicted CVR in women, and estradiol did not show stress-sensitive effects. These data provide evidence that increased vascular reactivity may be one mechanism linking hostility to increased cardiovascular mortality in men and support the notion that hostility may have different implications for CVR in women. PMID- 16250732 TI - Differential responsivity of monocyte cytokine and adhesion proteins in high- and low-hostile humans. AB - This study tested the general hypothesis that high- and low-hostile respondents would show different patterns of change in monocyte cytokine and adhesion protein (MCAP) expression in response to pharmacologically induced alterations in sympathetic nervous system (SNS) and parasympathetic nervous system (PNS) balance. On 3 separate days, 4 high- and 4 low-hostile respondents received isoproterenol infusions after saline, atropine (PNS blockade), or neostigmine (PNS stimulation) pre-treatment. Dual color flow cytometry with fluorescently labeled monoclonal antibodies to CD 14 (monocyte market), interleukin-1, leukocyte function activator (LFA-1), Class II major histocompatibility complex (MHC-II), and tumor necrosis factor was used to quantify cytokine and adhesion protein expression on monocytes in blood samples drawn before and after the combination drug infusions on the 3 test days in each respondents. Following PNS stimulation and istoproterenol infusion there was a decrease (compared to saline pretreatment) in MHC II expression in high hostiles that was significantly (p<.02) different from an increase in low hostiles. A similar trend (p = .08) was seen for LFA-1 expression, with high hostiles showing an increase and low hostiles a decrease. These findings support the broad hypothesis that high-and low-hostile respondents will show different MCAP responses to pharmacologically induced alterations in SNS-PNS balance. Such differences could contribute to accelerated atherogenesis among high-hostile individuals. PMID- 16250733 TI - Sensitization, somatization, and subjective health complaints. AB - More than half of the days lost due to sickness absence are due to diagnostic groups that solely or mainly depend on subjective statements from the patient The most frequent subjective health complaints are musculoskeletal pain. These conditions do not seem to qualify as psychiatric or mental disorders, but are not strictly somatic states either. Terms like somatization may be inadequate terms for states that may be best understood as psychobiological feedback loops. Subjective health complaints is suggested as a neutral, descriptive term. Only a minority requires treatment and sickness compensation for prolonged periods for these very common states. In these patients the neurons in feed-forward and positive feedback loops may have developed sensitization. These patients tend to show an abnormal sensitivity to sensory input from muscles, the gastrointestinal tract, and to smell and taste. It seems to be futile to search for single-factor solutions. This approach opens up for the possible effectiveness of many different types of treatment, breaking the feedback loops. PMID- 16250734 TI - Risk for hypertension and pain sensitivity in women. AB - To evaluate the possible negative association between risk for hypertension and pain sensitivity by in women 24 healthy young adult women with a parental history of hypertension and 24 without a parental history of hypertension participated in 2 laboratory sessions in which electric shock and the cold-pressor test were administered To assess the possible role of stress-induced analgesia in blood pressure-related hypoalgesia, the sessions were identical with the exception of the fact that participants were exposed to a stressful 20-min videogame before the pain stimuli on one occasion and a nonstressful control task on the other. Women with a parental history of hypertension and high blood pressure reactivity to the videogame displayed a reduced sensitivity to electric shock on both days, suggesting that risk for hypertension is associated with reduced sensitivity to at least some pain stimuli in women. Blood pressure reactivity to stress associated baroreceptor stimulation was not implicated as a mediator of decreased pain perception. However, other analyses revealed an effect of family history on shock pain only among women who report relatively high anxiety, suggesting that other aspects of the stress response may be involved in this phenomenon. PMID- 16250735 TI - Directive and nondirective social support in diabetes management. AB - Directive and Nondirective Support were distinguished (interrater agreement = 88.2%) through open-ended interviews completed by 60 adults with Insulin Dependent Diabetes Mellitus. Supporting validity, the sum of both Directive and Nondirective Support was correlated with scores on the Interpersonal Support Evaluation List (ISEL; r = .36). Supporting their distinction, Directive and Nondirective Support were inversely correlated (r = -.26), and Directive but not Nondirective Support decreased with age. Partial/correlations controlled for general support as measured by the ISEL, to examine the unique associations of Directive and Nondirective Support. For those less than 30 years old, Nondirective Support was associated with better metabolic control (p = .004). For those 30 or older, Directive Support appeared counterproductive, being associated with greater negative mood (p = .02). Different types of support may play different roles in different areas of disease management (e.g., metabolic control versus mood) and as a function of individual characteristics such as age. PMID- 16250737 TI - Salivary lysozyme: a noninvasive marker for the study of the effects of stress of natural immunity. AB - Salivary lysozyme levels were measured to determine the effect of stress on innate mucosal immunity. In the first study perceived stress levels of 39 participants were using a stress/arousal checklist questionnaire. Salivary lysozyme levels were found to show a negative correlation (r = -.477, p < .01) with the perceived stress level. In a second study, salivary lysozyme concentrations were measured during periods of high and low stress in the form of an actual stressor--final year examinations. The concentration of lysozyme was found to be significantly (p < .05) lower in the sample taken before the examination when the students were awaiting entry into the examination hall compared to the levels after completion of all examinations. These results indicate that salivary lysozyme concentrations are sensitive to psychological stress and could be utilized as a potential marker in studies looking at the effects of stress on immunity. Because the usefulness of sIgA as a marker has been questioned, the use of salivary lysozyme as a noninvasive index of mucosal immunity warrants further investigation. PMID- 16250736 TI - Relations of plasma ACTH and cortisol levels with the distribution and function of peripheral blood cells in response to a behavioral challenge in breast cancer: an empirical exploration by means of statistical modeling. AB - This study explores by means of statistical modeling the relations between adrenocorticotrophin hormone (ACTH) and cortisol levels and distribution and function of peripheral blood cells in response to an acute stressor consisting of a standardized speech task in breast cancer patients with axillary lymph node metastases and distant metastases. As a control group age-matched women participated in this study. The preliminary findings show that the effect of ACTH on immunoreactivity is related to the health of the doctor. In node-positive breast cancer patients and healthy women, ACTH has a modest positive effect on T lymphocyte percentages and on pokeweed-induced proliferation at baseline and in response to the speech task. In contrast, in breast cancer patients with distant metastases, ACTH has a negative effect on T lymphocyte and function at baseline and in response to the stressor. Interestingly, neither ACTH nor cortisol levels were related to natural killer (NK) cell percentages and natural killer cell activity (NKCA). In addition, it appeared that cortisol had a positive effect on CD3 cell percentages when the health of the donor was taken into account. This effect was most distinct on CD3 cells measured at baseline. If replicated on a larger scale, these findings may indicate that the hypothalamic pituitary adrenal axis plays a role in the adaptation of the host defenses in reaction to acute stress, particularly those involving T lymphocytes. Moreover, these findings may suggest that the health of the donor may be an important effect modification factor in the relations between neuroendocrines and immunoreactivity. PMID- 16250738 TI - The influence of dietary cholesterol on cardiac and hepatic Beta-adrenergic receptors in egyptian sand rats. AB - We examined the effects of dietary cholesterol on cardiac and hepatic beta adrenergic receptor functioning. Age-matched adult desert rodents (Psammomys obesus) were randomized to either a 5% cholesterol diet (CD, n = 20), or normal rabbit chow (RC, n = 18). After a 2-month exposure to the diets, animals were sacrificed and tissue from both heart and liver were retained for radioligand bindings studies. In heart tissue, cholesterol fed animals, relative to controls, showed an increased production of adenosine 3,5>-cyclic monophosphate (cAMP) in response to isoproterenol. Cholesterol supplementation was not associated with an increase in heart beta-adrenergic receptor number. Animals fed the 5% cholesterol diet showed significant increases in the number of beta-adrenergic receptor sites in hepatic tissue (M = 13.2 vs. 10.4 pmol/mg protein, CD and RC, respectively). The increased number of receptor sites in the liver was accompanied by a significant increase in isoproterenol-stimulated cAMP production. Results are supportive of the hypothesis that dietary cholesterol contributes to an upregulation of beta-adrenergic receptor function in cardiac, as well as hepatic tissue. These findings may be relevant to the observations of excessive stress induced cardiovascular reactivity in persons with high cholesterol levels. PMID- 16250739 TI - Relation of mood ratings and neurohormonal responses during daily life in employed women. AB - Diurnal variations in urinary norepinephrine, epinephrine, cortisol, and self reported mood states were examined in 101 employed women. Urine was collected on 2 consecutive workdays at 3 time periods: (1) overnight, (2) daytime, and (3) evening. Self-reports of 14 mood states were combined to correspond with the urinary collection periods. Factor analyses revealed 3 mood factors: Pressured (rushed, busy, stressed, hassled, and tense), Distressed (afraid, depressed, bored, and nervous), and Contented (satisfied, thoughtful, excited, and calm). The Distressed factor was significantly associated with norepinephrine excretion (p <.001). The Pressured factor was significantly associated with cortisol, epinephrine, and norepinephrine excretion (p <.001) independent of age, ethnicity, marital status, parental status, department within the company and activity level. Secretion of urinary hormones and the Pressured factor followed the same diurnal pattern. Specifically, as women's self-ratings of feeling pressured initially increased and then decreased over the course of the workday, urinary hormones also peaked and then dropped. Once time of day was statistically controlled, however, the mood factors were no longer significantly related to the urinary hormones. PMID- 16250740 TI - Blood pressure, gender, and parental hypertension are factors in baseline and poststress pain sensitivity in normotensive adults. AB - We studied 38 men and 36 women to learn whether a brief speech stressor reduced normotensive humans' thermal pain sensitivity, whether baseline and poststress pain threshold and tolerance varied with blood pressure (BP) and hemodynamic measures, and whether these relations differed by gender and parental hypertension (PH). PH-women with low-resting BPs had lower baseline pain tolerance than did all the other groups (ps <.05), and this group alone exhibited stress-induced analgesia (p = .008). In women, pre- and poststress pain tolerance varied directly with rest and stress BP (ps <.05). PMID- 16250741 TI - Relation of chronic and episodic stressors to psychological distress, reactivity, and health. AB - This study tested the hypothesis that exposure to chronic stressors, which reflect persistent, negative life situations, would have greater physiological, psychological, and physical health costs than exposure to episodic or intermediate-length stressors, which reflect relatively transient, negative life situations. We also tested whether cardiovascular reactivity, conceptualized as a marker of underlying pathophysiological states, would mediate the relation between chronic stress and psychological distress and illness. Participants were 75 male and 75 female college students. Compared with students experiencing few chronic life stressors, students experiencing many chronic life stressors had exaggerated cardiovascular responses to acute challenges, delayed recovery to resting levels of cardiovascular functioning after the acute challenges, elevated psychological distress levels, and they reported more illnesses. None of the outcomes was associated with the number of episodic or intermediate-length life stressors students experienced. Cardiovascular reactivity did not mediate the stress-distress or stress-illness associations. The results suggest that ongoing stressors that are static are more detrimental to health and well-being than are episodic of change-related stressors. PMID- 16250742 TI - Predictors of exercise adherence following participation in a cardiac rehabilitation program. AB - Patients who participate in cardiac rehabilitation programs (CRP) experience significant improvements in quality of life, rehospatilization rates, and mortality associated with cardiovascular disease. The potential efficacy of CRP is limited however, by significant program dropout rates and poor patient adherence to prescribed exercise regimens following rehabilitation. Recently, models of motivational readiness for behavior change, such as the Transtheoretical Model, have been applied to understanding the process of exercise adoption and maintenance. Interventions based on this dynamic model of behavior change have produced significant improvement in adherence to exercise in community and worksite populations. This study investigates the applicability of this model to this special population. Sixty-two men and women completed measures of motivational readiness, self-efficacy, and decisional balance for exercise adoption upon entry into a 12-week cardiac rehabilitation program at posttreatment and at a 3-month follow-up. Patients made significant gains during CRP in time spent exercising and self-efficacy, but not in utilization of the cognitive processes or in the perceived benefits of exercising. Exercise maintenance at follow-up was differentially associated with self-efficacy, decisional balance and use of behavioral processes at posttreatment. Motivation based models of exercise adoption may provide insights regarding the adoption and maintenance of regular physical activity in cardiac rehabilitation populations. PMID- 16250743 TI - Modification of physical activity 5 months after myocardial infarction: relevance of biographic and personality characteristics. AB - The relation between modification of physical activity, a risk factor for coronary heart disease, and personality characteristics was assessed in 166 survivors of a first myocardial infarction (MI). Physical activity was assessed before MI in retrospect and again 5 months after MI. Patients were divided into 3 categories according to their current daily-life physical activities: less active than before MI (n=24), equally active as before MI (n=82), or more active than before MI (n=60). A significant differentiation was found between patients who became less physically active than before MI and the other 2 categories. This less active category was characterized by feelings of disability, a low level of vigor, and feelings of anxiety. In addition, this patient group was on average older and more often female. The results were adjusted for participation in a cardiac rehabilitation program. Finally, the discussion recommends involving psychological intervention in the exercise program for the less active category of patients to diminish feelings of anxiety and disability and to improve vigor. PMID- 16250744 TI - You want to measure coping but your protocol's too long: consider the brief COPE. AB - Studies of coping in applied settings often confront the need to minimize time demands on participants. The problem of participant response burden is exacerbated further by the fact that these studies typically are designed to test multiple hypotheses with the same sample, a strategy that entails the use of many time-consuming measures. Such research would benefit from a brief measure of coping assessing several responses known to be relevant to effective and ineffective coping. This article presents such a brief form of a previously published measure called the COPE inventory (Carver, Scheier, & Weintraub, 1989), which has proven to be useful in health-related research. The Brief COPE omits two scales of the full COPE, reduces others to two items per scale, and adds one scale. Psychometric properties of the Brief COPE are reported, derived from a sample of adults participating in a study of the process of recovery after Hurricane Andrew. PMID- 16250745 TI - Behavior and metabolic disease. AB - The link between behavioral factors and disease is not well-defined. Although connections between a fight-flight reaction to environmental stress and hypertension have been much discussed, a potential disease association to a defeat-type of reaction has been much less considered. This is characterized by an elevated activity of the hypothalamo-pituitary-adrenal (HPA) axis, which is difficult to measure over a sufficiently long period of time. There is now considerable evidence that the characteristic peripheral endocrine abnormalities following a chronic HPA axis activation is directing storage fat to central, visceral adipose tissue depots. This evidence come from detailed molecular and cellular studies, clinical observation, and intervention trials, as well as from statistical associations between visceral fat accumulation and HPA axis activation in a number of conditions. Central fat accumulation measured conveniently as the waist/hip circumference ratio (WHR), is therefore probably a surrogate measurement for a chronic or repeated activation of the HPA axis. The WHR consequently provides a possibility to examine connections between environmental factors resulting in a hyperactive HPA axis, which is a consequence of a defeat-type of reaction to perceived stress. Such statistical associations have been examined in several population samples. The WHR has been found to be linked to a number of psychosocial and socioeconomic handicaps among both men and women, as well as to traits of psychiatric disease and use of alcohol and tobacco. Measurements of moderate obesity without WHR elevation often show reverse relations. It is suggested that measurements of central fat distribution such as the WHR may be used as a surrogate for chronic or repeated HPA axis activation, a consequence of a defeat-type of reaction to perceived environmental stress. This may provide a novel, convenient method to trace adverse bodily consequences of environmental stress leading to disease. This is also suggested by the fact that the WHR is now an established, unusually powerful risk factor for several prevalent diseases, which were previously suggested to have links to psychosocial and socioeconomic handicaps. PMID- 16250746 TI - Elderly hearing-impaired persons' coping behavior. AB - Little is known about the coping behavior associated with hearing impairment. In this study, 24 elderly hearing-impaired participants participated in a structured video interview including communication faults (provocations) presented in association with sounds. The participants also completed measures of self perceived hearing handicap, optimism, and depression. Several behaviors were coded independently by two observers using a rating scale that reached average Cohen's kappa of .75. A more conservative estimate resulted in a kappa of .50. Results showed that the provocations affected behavior differently. Especially problematic for the participants was a provocation in which the interviewer talked with his hand in front of his mouth. With one exception, no gender differences were found. Few significant relations between the assessed behaviors and self-assessed handicap, optimism, and depression were found. Hearing measurement (pure tone average and speech recognition) were not clearly related with the coping behaviors assessed. Behavioral observation techniques designed for the elderly hearing impaired should be further studied. PMID- 16250747 TI - Is there an association among low untreated serum lipid levels, anger, and hazardous driving? AB - Anger and hazardous driving were explored in two studies for their possible explanatory role in the low cholesterol-violent mortality association. In Study 1, we obtained fasting blood samples and indices of anger and driving habits from 102 healthy young adults. Among men (but not women), lower low-density lipoprotein (LDL) cholesterol levels were associated with significantly higher anger (r = 41, p < .01) and unsafe driving habits (r = -.30, p < .05), and although total cholesterol levels were correlated in the predicted direction, these were not significant. In Study 2, 78 healthy young men completed the same measures, and this time, both lower total and LDL-cholesterol were significantly related to increased anger and dangerous driving. Implications of these findings are discussed as are possible avenues for future research. Further scrutiny of the components of serum cholesterol and their relation to psychological factors may aid us in better understanding the reasons why men are at increased risk for hazardous behavior and violent death. PMID- 16250748 TI - Relation between beta-adrenergic receptor density and lymphocyte proliferation associates with acute stress. AB - The relations between acute changes in beta-adrenoreceptor density on lymphocytes and lymphocyte proliferation to a laboratory speech stressor were examined. Male participants either prepared and delivered a challenging speech task or read from a list of words. Participants engaged in the speech task, but not those involved in reading, exhibited significant increases in systolic blood pressure (an index of sympathetic nervous system arousal) and beta-adrenoreceptor density, accompanied by significant decreases in lymphocyte proliferation to pokeweed mitogen (PWM) but not concanavalin A (Con A). Increases in adrenoreceptor density significantly predicted decreases in lymphocyte activity in response to both mitogens. The increases in beta-adrenoreceptor density in the speech task condition occurred within 5 min after the baseline period, when participants were preparing for the speech task, but not yet speaking. These findings suggest that an acute stressor eliciting sympathetic nervous system activity can evoke rapid changes in beta-adrenoreceptor activity and decreases in mitogenic responses. PMID- 16250749 TI - The differential effects of employment status on chronic pain and healthy comparison groups. AB - We evaluated the differential effects of employment status on chronic pain and healthy comparison groups. Forty unemployed and 43 employed individuals with chronic pain, as well as 43 unemployed and 45 employed healthy comparison participants completed a series of measures assessing background information physical and psychological adjustment, and psychosocial features of employment status. In general, participants experiencing both chronic pain and unemployment reported poorer adjustment than the other groups and more financial strain, less structured and purposeful activity, fewer opportunities for skill use and task variety, and decreased social support than the employed chronic pain and healthy comparison groups. Groups did not differ, however, in work ethic values. Scores on measures of pain severity and features of employment status accurately predicted the group membership of more than 70% of respondents from four groups. The study suggests that there is utility in understanding experiences of chronic pain patients on the basis of features of their current employment status and points to a need for multidimensional measures that evaluate psychosocial facets of employment and unemployment specifically for chronic pain samples. PMID- 16250750 TI - The effect of labor force participation on coronary heart disease risk factors among middle-aged women: a cross-sectional study in a Japanese rural district. AB - This study examined labor force participation as a risk factor for coronary heart disease. The participants were 224 women aged 45 to 64, residing in a rural district in Japan. The working women spent a significantly greater number of working hours (paid and unpaid) and had a more "Westernized" food preference than homemakers. After adjusting for physical, psychosocial, and behavioral covariates, working women had significantly lower systolic blood pressure and blood glucose levels than homemakers. The "healthy worker effect" did not explain these differences. Middle-aged rural Japanese women who worked outside the home were slightly healthier than homemakers, despite their less healthy lifestyle. Total working hours were positively associated with systolic and diastolic blood pressures and total cholesterol in multiple regression analyses, which suggests that work overload has a harmful effect. A more suitable social environment is required if f Japanese working women are to enjoy a healthier life. PMID- 16250751 TI - The challenges of future behavioral medicine. AB - To a large extent, behavioral medicine originates from the United States, and more specifically from the scientific traditions of pragmatism and behaviorism. The core notion of individual learning, mastery, and development embedded in these traditions has lent support to an almost exclusive concern with individual behavioral modification. However, individual mastery and welfare are increasingly threatened by powerful adverse socioeconomic and sociocultural developments, especially by growing social inequalities in health and by expanding social disintegration. Social differentials in morbidity and mortality are documented even in the most economically advanced countries where health-damaging lifestyles (e.g., cigarette smoking, diet) and stressful conditions of relative deprivation in occupational life and elsewhere contribute to the observed pattern. In addition, detrimental effects on health produced by social disintegration are manifest, most notably in societies that undergo rapid socioeconomic change. Implications of these developments for future behavioral medicine are discussed at the level of scientific analysis and of preventive and therapeutic intervention. PMID- 16250753 TI - Association between type A behavior pattern and coronary artery spasm in Japanese patients. AB - The relations between the Jenkins Activity Survey (JAS) Type A score and coronary atherosclerosis and spasm were examined in 192 patients (115 men and 77 women) undergoing coronary angiography. Thirty-nine men (34%) and 12 women (16%) had significant (>or=75%) coronary stenosis. In 97 patients (54 men and 43 women) with no significant coronary stenosis, a coronary spasm provocation test by 0.2 to 0.4 mg of ergometrine was performed, by which a significant focal spasm (>or=75% reduction of luminal diameter) was induced in 22 men (40%) and 5 women (12%). Men who showed a significant focal spasm had a higher Type A score than men who showed no such spasm (1.9 +/- 8.4 vs. -2.4 x 8.3, p < 0.05). Discriminant analysis in male patients revealed that the induction of coronary spasm can be predicted by the JAS Type A score and smoking habit (p = 0.04). No such association was found for female patients. These results suggest that the Type A behavior pattern as assessed by the JAS may be associated with coronary spasm in Japanese men without significant coronary stenosis. PMID- 16250752 TI - Hormone replacement therapy: knowledge, attitudes, and well-being among middle aged Australian women. AB - Use of hormone replacement therapy (HRT) is increasing, yet little is known of women's perceptions of HRT. A telephone survey examined relevant knowledge and attitudes, and psychological well-being, in 258 women aged 51 to 60 (111 HRT users, 47 previous users, and 100 never-users). Although HRT users were better informed than nonusers, knowledge was generally low. Half the HRT users could not name any hormone used in HRT, and one third could not give any reason for using HRT. Current users, however, had more positive attitudes to HRT. Groups defined by HRT usage did not differ on well-being or current symptomatology, although current and previous HRT users reported having previously experienced a higher level of symptomatology than never-users, suggesting that HRT may have provided symptom relief for those women who chose to use it. In contrast to previous research, women with a history of hysterectomy did not show more emotional distress than others. Results suggest that Australian women frequently make choices concerning HRT without adequate knowledge, and that HRT may reduce symptoms but may have little impact on psychological well-being. Optimal usage of HRT by middle-aged women will rely on a clearer understanding, both of its effects and of women's attitudes toward its use. PMID- 16250754 TI - The ability of active versus passive coping tasks to predict future blood pressure levels in normotensive men and women. AB - Casual blood pressure (BP) after a 2-year follow-up interval was determined in 40 normotensive men and women (20 Blacks and 20 Whites), who had been initially tested for cardiovascular responses to a variety of active and passive coping tasks, including active speech, passive speech, reaction time, and forehead cold pressor tasks. Stepwise multiple regression analyses were used to identify the best model for predicting follow-up BP. Average systolic blood pressure (SBP) level during cold pressor stress was the single most powerful predictor of casual SBP over 2 years even after controlling for initial resting SBP. Other predictors of follow-up SBP were initial SBP, parental history of hypertension, and heart rate and SBP during passive speech (final model R(2) = .78). For follow-up diastolic blood pressure (DBP), the only significant predictors were initial DBP and male gender. These results contribute to a growing body of literature that suggests that cardiovascular measures observed during stressors have predictive validity above and beyond that of traditional predictor variables. PMID- 16250755 TI - Variations in plasma lipid concentration during examination stress. AB - The effect of psychological stress on plasma lipids was studied in 40 law students (20 men and 20 women). Plasma cholesterol, triglyceride, and apolipoprotein concentrations were examined at the beginning of the quarter and during the week before final examinations. Cortisol, blood pressure, and heart rate, and self-reports of stress and workload were also measured to verify that examinations were associated with increased stress levels. Perceived stress, perceived workload, and cortisol increased before examinations. Low density lipoprotein cholesterol (LDL-C) increased 5.8 +/- 13.9 mg/dL, and apolipoprotein B (apo B) increased 2.9 +/- 4.0 mg/dL. High density lipoprotein cholesterol decreased in women only. These changes were not due to changes in dietary intake or indexes of plasma volume. However, changes in cortisol and changes in LDL-C and apo B were associated, suggesting a neuroendocrine component to the effects. These results suggest that episodic, stressful situations may lead to potentially atherogenic changes in lipid and lipoprotein concentrations. PMID- 16250756 TI - Secretory immunoglobulin A and cardiovascular responses to acute psychological challenge. AB - Cardiovascular activity and secretory immunoglobulin A (sIgA) in saliva were recorded at rest, during a 30-min computer game task, and during subsequent recovery. Blood pressure (BP) rose and remained elevated during the task and returned to resting levels during recovery. This pressor response was produced by increased total peripheral resistance rather than increased cardiac output. SIgA secretion rate also increased during the task, although the effect proved significant only toward the end of the task. As such, the data provide preliminary indication that sIgA is sensitive to acute psychological challenge in the laboratory. Although correlational analyses revealed that sIgA reactions were stable, they were not significantly correlated with pressor reactions. The influence of task uncertainty was explored by comparing individuals who had previously played the computer game with those who had not. Task-induced increases in BP and sIgA were a feature of individuals new to the computer game. In contrast to these novice players, experienced players showed minimal increases in BP and no increases in sIgA. PMID- 16250757 TI - Depressive symptoms and disability in acute and chronic back pain patients. PMID- 16250758 TI - Psychometric qualities of the RAND 36-Item Health Survey 1.0: a multidimensional measure of general health status. PMID- 16250759 TI - Relations among night work, dietary habits, biological measure, and health status. AB - A cross-sectional study was conducted to investigate dietary intake, behavioral habits, and clinical and metabolic differences in night workers compared to day workers and to evaluate the metabolic differences associated with diet and body habits that occur between these two groups. Dietary habits, biological parameters, and health status were collected in 1,200 night workers and in an equal number of day workers, matched for gender, age, and socioeconomic status. Our findings demonstrated that night workers had poorer dietary habits and metabolic profile compared to day workers with a similar overall health status. These differences were associated with a higher prevalence of some cardiovascular risk factors such as smoking and obesity. PMID- 16250760 TI - Patterns of immune, neuroendocrine, and cardiovascular stress responses in asymptomatic HIV seropositive and seronegative men. PMID- 16250761 TI - The psychobiology of hostility: possible endogenous opioid mechanisms. AB - This study examined the role of endogenous opioids in the relation between hostility and cardiovascular stress responsiveness. Forty-six men completed the Cook-Medley Hostility Scale and experienced a laboratory pain stressor once under opioid blockade and once under placebo. Hostility scores were significantly related to the magnitude of change in cardiovascular reactivity/recovery resulting from opioid blockade. Low scorers on the Cynicism subscale displayed increases in heart rate (HR) reactivity under blockade relative to placebo, with reactivity decreases noted in high scorers. Low Hostile Affect scores were similarly associated with impaired diastolic blood pressure recovery under opioid blockade. HR recovery results were somewhat different, with high scorers on Aggressive Responding and the total Cook-Medley displaying improved HR recovery under opioid blockade, with no change noted in low scorers. These data provide preliminary support for the hypothesis that low hostile individuals rely on endogenous opioids for buffering cardiovascular stress responsiveness, but high hostiles do not. PMID- 16250762 TI - Differential hostility profiles accompany different hemodynamic response patterns. AB - Cardiovascular reactivity has been proposed as a mechanism by which psychosocial variables may lead lo coronary heart disease. Although reactivity as a generalized response has been linked lo psychological factors, there are specific hemodynamic profiles that underlie blood pressure reactivity. This study characterized subsets of young adults as myocardial or vascular reactors, in response to three laboratory tasks: mental arithmetic, videogame, and anger recall interview. Vascular reactors had higher diastolic blood pressure increases to all tasks, whereas reactor groups were equivalent at rest and on systolic blood pressure responses. Correlational analyses were conducted to determine whether psychological dimensions are uniquely associated with myocardial or vascular reactors' cardiovascular responses, at rest and during acute stressors. For myocardial reactors, resting systolic blood pressure was higher in hostile and suspicious individuals, whereas reactivity was linked to impulsivity. For vascular reactors, low trust, low gregariousness, and high depression were associated with diastolic reactivity to tasks. The predictive validity of hostility, distrust, sociability, and depression for cardiovascular pathology may vary with reactor type. PMID- 16250764 TI - The placebo effect in pain reduction: the influence of conditioning experiences and response expectancies. AB - We investigated the role of conditioning experiences and response expectancies in the generation of placebo effects. On 3 sequential days (Test 1, Experimental Session, Test 2), 66 female undergraduates were presented with a series of pain stimuli. For the experimental group, placebo administration (analgesic cream) was paired with a decrease in the painful stimulus. Two control groups were used to explore the relative contributions of verbally induced expectancies and contingent unconditional stimulus experiences per se. The results show that placebo-induced pain reduction can he obtained as a result or a conditioning procedure, independent of verbally induced expectancies. Mere verbal persuasion was not sufficient to elicit placebo-induced pain reduction. Irrespective of the experimental manipulations, the placebo effect was related to both reduced pain expectations and reduced fear of pain. Although conditioned placebo responses were evident at the subjective level, no placebo effects emerged at the physiological level. PMID- 16250763 TI - Are stress-induced immunological changes mediated by mood? A closer look at how both desirable and undesirable daily events influence sIgA antibody. AB - This investigation tested a three-path model of mood as a mediator of the relation between stress and immunity. Seventy-two married men completed end-of day diaries in which they rated their mood for that day and the desirability of the day's events for I2 weeks. Events were coded as either desirable (nonstressful) or undesirable (stressful) in nature. Immunological functioning was assessed by secretory immunoglobulin-A NgA) antibody response lo an oral antigen. Regression analyses indicated that negative mood partially mediated the immunological response to both undesirable and desirable events. Undesirable events lowered antibody levels primarily by increasing negative mood: desirable events increased antibody levels by decreasing negative mood. Evidence for mediation by positive mood beyond that found for negative mood was weak. PMID- 16250766 TI - Effect of the number of high-fat and low-fat cues on food choice. AB - This study examined the effects on food choice of increasing the number of healthy items available (fruit) and decreasing the number of unhealthy items available (candy bars). A similar choice, involving nonfood items, was also examined. Two hundred eighty-nine men and women were randomly assigned to 1 of 4 experimental groups: (a) control group, (b) increased number of fruits, (c) decreased number of candy bars, and (d) combination. Between 30% and 40% of participants chose fruit regardless of the amount of fruit and candy presented: there was no effect of increasing fruit or decreasing candy bars. However, restrained participants and current dieters were more likely to choose fruit. In contrast, both stimulus control techniques were effective in increasing the percentage of participants choosing a nonfood item. These results suggest that stimulus control may not be sufficient to modify food choice: other powerful factors affect eating behavior, and these must be considered. PMID- 16250765 TI - Predicting subjective disability in chronic pain patients. AB - Subjective disability is considered as the variable that reflects the impact of chronic pain on a patient's life. This study examines the questions of which syndrome or patient characteristics determine subjective disability and whether there are differences between samples of patients with chronic headaches and low back pain. Direct pain variables and depression, pain coping strategies, and pain related self-statements (including catastrophizing) are introduced into multivariate regression analyses as potential predictors of disability using a sample of 151 pain patients. Disability is not predicted by pain severity in patients with headaches or back pain. Psychological variables, especially coping strategies, are far more influential. Coping explains more variance in disability in the headache sample than in the chronic law hack pain group, whereas depression is more relevant for the degree of disability in the back pain sample. In this study, we present a critical analysis of possible interpretations of our results. We point to an overlap of concepts underlying some of the variables used: this overlap also considerably invalidates conclusions drawn from a multitude of studies done in this field, including the one presented. We strongly argue for a conceptual clarification, and consequently for the revision of assessment instruments, before further empirical work in this area is done. PMID- 16250767 TI - Aerobic and resistance exercise training effects on body composition, muscular strength, and cardiovascular fitness in an HIV-1 population. AB - Although HIV-1 infection rates peaked in the 1980s in the United States, advanced stage HIV disease will grow by about 40% in the next 40 years. This fact has signaled the need for intervention strategies that go beyond primary prevention and into treatment of the complications associated with this chronic disease. Recently, the role of exercise in reversing the wasting process experienced by individuals with HIV-1 has received much needed attention. The purpose of this study was to examine the independent contributions of a 12-week aerobic or resistance weight training exercise regimen in improving body composition, muscle strength, and cardiovascular fitness in an HIV-1 population. Participants for this study (N = 33) were randomly assigned to an aerobic exercise training group, a resistance weight training group, or a stretching/flexibility control group. Measures of body composition, muscular strength, and cardiovascular fitness were assessed both prior to and following completion of the intervention. In general, the results revealed significant improvements in the amount of lean muscle tissue, upper and lower body muscular strength, and predicted VO(2)max for exercise participants, whereas control participants experienced declines in all physiological measures. Based on these findings, it is suggested that exercise may be one complimentary therapeutic modality capable of combating the wasting process associated with advanced HIV-1 infection. PMID- 16250768 TI - Does the stressed patient with chronic fatigue syndrome hyperventilate? PMID- 16250769 TI - Anger expression and lipid concentrations. AB - We used 2 different strategies to examine the relation between anger expression and lipid concentrations in 116 middle-aged men. Using the common analytic method used in the literature, the group crossing approach, we examined whether Anger In. Anger-Out, and their interaction were related to lipids. Regression analyses revealed that Anger-In and Anger-Out were marginally related to total cholesterol. These associations disappeared after controlling for hostility, anger, and anxiety. Using a new intraindividual difference approach, we determined individuals' relative dominance of Anger-In and Anger-Out and examined linear and quadratic associations with lipids. Regression analyses revealed the quadratic was related to both total cholesterol and low-density lipoprotein cholesterol (LDL-c), indicating that individuals who almost always express their anger or almost never express their anger had both elevated total cholesterol and LDL-c. The curvilinear association with total cholesterol persisted even after controlling for hostility, anger, and anxiety. PMID- 16250770 TI - Effects of academic examination stress on eating behavior and blood lipid levels. AB - The influence of academic examination stress on eating behavior and lipid profiles and the moderating effect of dietary restraint, trait anxiety, and social support availability was assessed in university students. One hundred and seventy-nine students were divided into exam-stress groups (51 women, 64 men) and control groups (48 women, 16 men) and were assessed at baseline and then within 2 weeks of exams or an equivalent point for the control group. Perceived stress, emotional well-being, and fasting lipid profiles were measured, and dietary information was collected by interview. The exam-stress group reported significant increases in perceived stress and deterioration in emotional well being at the exam sessions compared with baseline sessions. No general effects of exam stress on food intake were observed, and there was no interaction between stress and dietary restraint. However, students in the exam-stress group with high trait anxiety and low social support showed significant increases in total energy intake between baseline and exam sessions, whereas individuals with low trait anxiety and high social support showed a reduction in energy intake. Students with high trait anxiety and low social support showed increases between baseline and exam sessions in the amount of fat and saturated fat consumed. Women in the exam-stress group taking oral contraceptives showed a significant increase in total cholesterol between baseline and exam sessions. The results are discussed in relation to the effects of naturally occurring episodic stress on health behavior and on lipid profiles. PMID- 16250772 TI - Parental history of hypertension and cardiovascular response to stress in Black and White men. AB - White offspring of hypertensives typically exhibit an elevated cardiovascular response to stress. Studies of Black offspring of hypertensives have been fewer, with inconsistent results. This may be due, in part. to incomplete characterizations of hemodynamic responses. This study examines cardiovascular reactivity in Black and White offspring of hypertensives with a particular focus on vascular resistance responses. A total of 62 healthy normotensive men, 41 with a parental history of hypertension (PH+: 21 Blacks, 20 Whites), and 21 without parental hypertension (PH-: 7 Blacks, 14 Whites) engaged in a series of laboratory tasks. Both Black and White PH+ participants exhibited elevated diastolic blood pressure (DBP) responses, but to different patterns of stressor tasks. Familial differences in total peripheral resistance response were also obtained for Black and White participants in a comparison across all tasks, but were particularly evident in tasks when PH+ participants had elevated DBP responses. These results suggest that a parental history of hypertension is an important moderator of cardiovascular, and in particular peripheral vascular, responses to stress in Black and White individuals. PMID- 16250771 TI - Effects of racist provocation and social support on cardiovascular reactivity in African American women. AB - It has been speculated that exposure to the chronic stress of racism contributes to the high rates of hypertension among African Americans. Social support may buffer the effects of stress on cardiovascular (CV) health by attenuating stress induced CV responses that have been linked to hypertension. In this study we investigated the effects of racism and social support on CV reactivity in African American women. Participants showed greater increases in CV and emotional responses while responding and listening to racist provocation. Augmented blood pressure (BP) persisted through recovery following racial stress. Participants receiving no support showed the greatest increases in anger during racist provocation. No significant effects were seen for support on CV reactivity. These results provide some of the first evidence that interactive confrontation with racism elicits significant increases in CV reactivity and emotional distress. Furthermore, individuals receiving less support may be at greater risk for the potentially health-damaging effects of racial stress. These findings may have significant implications for the health of African Americans. PMID- 16250773 TI - Assessment of job stress dimensions based on the job demands- control model of employees of telecommunication and electric power companies in Japan: reliability and validity of the Japanese version of the Job Content Questionnaire. AB - To investigate the reliability and validity of 4 selected scales from the Japanese version of the Job Content Questionnaire (JCQ. Karasek, 1985)-decision latitude, psychological demand, supervisor support, and coworker support-a survey was conducted on a total of 626 employees of telephone and electric companies in Japan. The survey questionnaire was composed of 22 items. Data from 472 male and 108 female respondents were analyzed. Cronbach's alpha coefficients for the four JCQ scales ranged from .61 to .89 for men and from .65 to .87 for women. Scree plots based on factor analyses of scale items indicated that one major factor explained 30% to 75% of each scale variance in men and women. Factor structures of the 22 items for men and women were consistent with those theoretically expected Distributions of the decision latitude scores among occupations for men and women were similar to those in the U.S. national samples; the scores significantly and positively correlated with occupational class. It is suggested that the JCQ scales are reliable and valid instruments for assessing job stressors in a Japanese working population. PMID- 16250775 TI - Psychosocial risk factors for cardiovascular disease in women: the role of social support. AB - Lack of social support is becoming increasingly important as a psychosocial risk factor in the study of coronary heart disease (CHD). There may also be an association between vital exhaustion and lack of social support. Because most research has focused on men, we decided to explore the associations between structural and functional social support and first myocardial infarction (MI) in women. Subjects were 79 women hospitalized with a First MI (mean age 59.3; SD = 9.3) and 90 women hospitalized with an acute surgical event (mean age 57.4: SD = 9 1). MI cases reported more vital exhaustion than did controls (p < ,040), and exhausted women reported less structural (p < .001) and functional support (p < .000). After controlling for age, hypertension, diabetes, menopausal status, smoking, nonanginal pain, and vital exhaustion, multiple logistic regression analysis showed that poor structural support was associated with an increased risk for MI. These results suggest that social support is associated with vital exhaustion, which is a well-known risk factor for CHD. Furthermore, it is suggested that women with only a few confidants are more at risk for MI even after adjustment for well-known risk factors for CHD. PMID- 16250776 TI - Family history of hypertension, exercise training, and reactivity to stress in rats. AB - In this study we sought to assess the role of exercise training on blood pressure (BP) reactivity to tailshock stress in rats with varying family histories of hypertension. Exercise training consisted of swimming 90 min per day in isothermic water for either 2, 6, or 10 months, beginning at 2 months of age. Control subjects were age-matched and did not exercise daily. Rats with either zero (Wistar-Kyoto), 1 (borderline hypertensive), or 2 (spontaneously hypertensive) hypertensive parents were studied. At the appropriate age, femoral artery catheters were implanted and rats were studied at rest and in response to a 20-min stress session. Exercise training reduced basal BP, especially in rats with a positive family history that were exercised for the longest duration. Reactivity to stress was actually significantly enhanced in trained rats. Thus, these data do not support the reactivity hypothesis, but suggest several reasons why the literature has been so inconsistent. The discussion emphasizes the importance of basal, rather than phasic. BP responses resulting from exercise training. PMID- 16250774 TI - Hemodynamic responses during psychological stress: implications for studying disease processes. AB - Investigation of the physiological correlates of psychological stress is of interest in relation to the putative impact of stress in the etiology of cardiovascular disease. Although the assessment of blood pressure and heart rate responses to psychological stress has been very informative, the addition of cardiac output measurement has added a further dimension to this research field. In recent studies, a more complete hemodynamic picture of the stress response has been documented in terms of cardiac output and systemic vascular resistance components of blood pressure changes. Different stressors have been shown to produce similar blood pressure increases due to quite different hemodynamic mechanisms. Furthermore, when faced with the same stressor, different individuals may exhibit pressor responses that are very different hemodynamically. There is growing evidence that these hemodynamic response patterns to psychological stress are stable individual traits. Response stability is a prerequisite for considering how stress-related hemodynamic changes may be implicated in the pathophysiology of cardiovascular diseases. Observations that hemodynamic response patterns in individuals at higher risk for the development of hypertension differ from those of lower risk individuals show that specific patterns of hemodynamic response are associated with disease processes. Although it is as yet unclear whether they represent markers or mechanisms. Overall. hemodynamic studies appear to he helping to refine our understanding of how stress can impact cardiovascular disease processes. PMID- 16250777 TI - Working despite pain: factors associated with work attendance versus dysfunction. AB - A cross-sectional investigation of psychosocial variables in 63 female employees matched for experienced pain was conducted to study the difference between back pain sufferers who were working (Copers) and those who were off work (Dysfunctional). The subjects reported moderate to severe pain often or always during the past year and were employed at the same hospital. Thirty-seven women who had not been off work for pain made up the Copers group, whereas 26 women who had been off work for their pain made up the Dysfunctional group. Subjects were interviewed and completed a battery of questionnaires designed to penetrate level of dysfunction, perceived health, work and social satisfaction, perceived workload, coping strategies, and pain beliefs. Multiple covariate analyses that controlled for perceived workload, smoking, low-back mobility, and obesity revealed significant differences between the groups on levels of functioning, pain beliefs, and coping strategies used. Dysfunctional subjects had stronger beliefs that pain was directly related to activities that they had little control over their pain, that their health was poor, and that they tended to focus more on their pain. A discriminant analysis correctly classified 83% of the subjects as to work status based on six psychosocial variables. These results not only demonstrate the importance of psychosocial factors in back pain, but underscore the fact that work absence for back pain may he controlled by psychological factors related to beliefs and coping strategies. Future research may attempt to use these factors in the screening of patients. PMID- 16250778 TI - Adrenocortical effects of caffeine at rest and during mental stress in borderline hypertensive men. AB - We examined the effect or dietary doses of caffeine (3.3 mg/kg, equivalent to 2 to 3 cups of coffee) on adrenocortical responses to behavioral stress in borderline hypertensive (BH) men using a randomized, double-blind, caffeine placebo crossover design. Cortisol levels were assessed in BH men and matched normotensive (NT) controls at rest and in response to 35 min of continuous work on a psychomotor task alternating with mental arithmetic. Caffeine at rest elevated cortisol among BHs hut not among NTs. Both groups showed significant cortisol responses to caffeine combined with the tasks. These findings may have implications for the dietary use of caffeine in persons at risk for hypertension when faced with stressful situations. PMID- 16250780 TI - On the relation among psychological distress, diabetes-related health behavior, and level of glycosylated hemoglobin in type I diabetes. AB - This study aimed at investigating the relation between psychological diabetes related health behavior, and metabolic aspects of diabetes. Fifty-one adult patients with type I diabetes mellitus took part in the study. Psychological status, health, and self-care behavior were assessed by means of questionnaires. Level of glycosylated hemoglobin (HbA(1c) served as the index of metabolic control. Depression was slightly elevated among women as was trait anxiety and blood-injury phobia or fear of medical interventions in all patients. Depression and anxiety were not related to duration of diabetes or presence of diabetes complications. As could be expected, patients who frequently checked their blood glucose level had a significantly lower level of HbA(1c) than those with infrequent checks. Patients with a marked blood-injury phobia carried out fewer daily checks of blood glucose level than those without, but blood-injury phobia was not directly related to HbA(tc) level. A higher level of HbA(tc) was, however, associated with mood deterioration. As depression was not related to health behavior, its effect on metabolic control is likely to be mediated via endocrine rather than behavioral variables. PMID- 16250779 TI - Modulatory effects of defense and coping on stress-induced changes in endocrine and immune parameters. AB - We examined whether habitual defense and coping affect the response of hormones (ACTH. cortisol, prolactin. endorphins, and noradrenaline) and immune parameters (numbers of T cells. B cells. natural killer [NK] cells, and proliferative responses to mitogens or antigens) to an acute laboratory stressor (i.e., solving a 3-dimensional puzzle and explaining it to a confederate) in 86 male high school teachers. Defense and coping were assessed by Kragh's tachistoscopic Defense Mechanism Test (a measure of perceptual defense) and by 4 questionnaire-based coping styles assessing instrumental mastery-oriented coping, emotion-focused coping, cognitive defense, and defensive hostility. The laboratory stressor per se caused a relative increase in immunological (in particular NK cells) and endocrine (cortisol, prolactin) parameters. Defense and coping, however, significantly codetermined the response to the stressor. In particular, instrumental mastery-oriented coping and perceptual defense were related to stress-induced changes in numbers of B cells and in the pituitary-adrenal hormones. The results indicate that the impact of a mild psychological stressor on the immune and endocrine system depends to a considerable extent on the specific ways people deal with stressors. PMID- 16250782 TI - Effects of high and low anxiety provoking instructions on the responses to the hyperventilation provocation test. AB - This study examined the effect of high and low anxiety provoking instructions in subjects submitted to a Hyperventilation Provocation Test (HVPT). Subjects were 43 out-patients referred to our clinic for a diagnostic examination of Hyperventilation Syndrome (HVS). Results showed that anxiety levels were affected by the instruction manipulation, but the magnitude of this effect was less than expected and the instruction manipulation had no effect on intensity arid type of reproduced symptoms, nor on symptom recognition. Subjects who met Diagnostic and Statistical manual of Mental Disorders (3rd ed., rev,; American Psychiatric Association, 1987) criteria for Panic Disorder (PD) were not more responsive to the instruction manipulation than non-PD patients. It is argued that the small effect of the manipulation is probably not due to the solidity of the HVPT but to the pervasiveness of pretest cognitions and expectations. In line with this, the report of HVS symptoms appeared highly related to psychological trait measures like anxiety, fear of bodily sensations, and a general tendency to report somatic symptoms. PMID- 16250781 TI - Reliability and validity of dyspnea measures in patients with obstructive lung disease. AB - Dyspnea, the clinical term for shortness of breath, is the primary symptom and an important outcome measure in evaluations of patients with lung disease. It is a subjective symptom that has proved difficult to quantify. Many dyspnea measures are available, yet it is difficult, based on the existing literature, to determine the most reliable and valid. In this study, we evaluated 6 measures of dyspnea for reliability and validity: (a) Baseline Dyspnea Index (BDI) and Transition Dyspnea Index, (b) UCSD Shortness of Breath Questionnaire (SOBQ),(c) American Thoracic Society Dyspnea Scale, (d) Oxygen Cost Diagram, (e) Visual Analog Scale, and (f) Borg Scale. Subjects were 143 patients (74 women) and 69 men) with obstructive lung disease, ages 40 to 86, FEV(1.0) 0.36 to 3.53 L, FVC 1.07 to 5.74 L. Dyspnea measures were assessed for test-retest reliability internal consistency, interrater reliability, and construct validity (i.e., correlations among dyspnea measures and correlations of dyspnea measures with exercise tolerance, health-related quality of life, lung function, anxiety, and depression). Results suggest that the SOBQ and BDI demonstrated the highest levels of reliability and validity among the dyspnea measures examined. PMID- 16250783 TI - Patterns of pain-relevant social interactions. AB - Pasts studied have focused on the individual effects of Solicitous, Distracting, and Negative spousal responses to pain on patient's pain behavior and pain severity. Because spouses may emit any combination of these responses, this research examined the conjoint effects of marital satisfaction and these perceived spousal responses by using WISE "step-down" procedure as described by Kenny and Judd (1983. Ninety-six married male chronic-pain patients completed the West Haven-Yale Multidimensional Pain Inventory (WHYMPI), Locke Wallace marital Adjustment Scale, and Pain Behavior Check List (PBCL). Final step-down models included significant 2-way and 3-way interactions on the PBCL measures of Distorted Ambulation and Seeking Help, significant main effects for Affective Distress, and a 4-way interaction on the WHYMPI Pain Severity scale. These trimmed models accounted for 23%: to 33% of the variance in the criterion measures. These results show that higher order interactions make unique contributions to the variance and should be examined along with main effects. PMID- 16250784 TI - Effect of trait hostility on cardiovascular responses to harassment in young men. AB - Hostile individuals may experience more extreme and frequent episodes of anger than nonhostile persons and thus may have exaggerated physiological responses to their environments. Such responses may be associated with increased risk of cardiovascular disease. This study examined cardiovascular responses of 24 low- versus 21 high-hostile young men to a mental arithmetic task administered with and without provocation in 2 x 2 (groups x Tasks) repeated measure design. hostility classifications were based on weighted interview ratings of Potential for Hostility. As predicted, high-hostile men showed differentially greater heart rate, blood pressure, and rate-pressure product changes only in response to the task administered with harassment (ps < .05). Similarly, high-hostile men reported more distress, tenseness, irritation, and greater concentration during the harassing task, compared to low-hostile men (ps < .05). Also, the harassing task elicited greater increases in vascular resistance and greater increases in stroke volume for all subjects, relative to the neutral task (ps < .003). Results suggest that hostile persons faced with anger evoking situations may produce a constellation of exaggerated cognitive-emotional and cardiovascular responses consistent with increased risk of cardiovascular disease. PMID- 16250785 TI - Anticipatory immune changes in women treated with chemotherapy for ovarian cancer. AB - Immune parameters were assessed in 22 women before chemotherapy for ovarian cancer and compared with assessment made at home 2 days earlier. In the hospital, as compared to home measures, patients had a lower percentage of lymphocytes and monocytes and a higher percentage of granulocytes. Absolute numbers of lymphocytes were lower at the hospital as compared to at home. T-cell proliferative responses to concanavalin A were elevated for cells isolated from hospital blood samples as compared to home samples. The observed changes in immune parameters were not related to levels o r state anxiety at home or at the hospital. The results show anticipatory immune changes in a group of patients receiving chemotherapy that did not include cyclophosphamide. PMID- 16250787 TI - Patient expectations and postoperative depression, anxiety, and psychosocial adjustment after temporal lobectomy: a prospective study. AB - This study examined the effects of patient expectations on short-term postoperative psychological adjustment alter surgery for epilepsy. Twenty-seven subjects (17 operative, 10 nonoperative controls) agreed to participate. A prospective longitudinal design assessed whether changes in psychosocial functioning had occurred at a mean of 7 weeks after surgery relative to the presurgical assessment. The study also examined the relation between postoperative psychosocial adjustment and preoperative patient expectations. It was found that seizure-free outcomes were associated with significant declines in depression and anxiety. Patients who exhibited preoperative optimistic expectations had lower levels of postoperative depression and anxiety, as long as they were rendered seizure-free. Conversely, those who had optimistic preoperative expectations but were not rendered seizure-free postsurgically continued to exhibit moderate to high levels of depression and anxiety. The theoretical significance of optimistic expectations and their effects on individual self-efficacy are discussed to support the notion that postsurgical adjustment is related to personal coping, even in those patients who are seizure free after surgery. PMID- 16250786 TI - Role of psychological stress in cortisol recovery from exhaustive exercise among elite athletes. AB - Life-event stress (LES) was used to classify elite athletes (n = 39) into high- and low-LES groups. A repeated measures analysis of variance revealed higher cortisol concentration after a graded exercise lest among the high-LES group relative to the low-LES group, which was maintained for up to 20 hr. Subsequent prospective analyses further indicated that high-LES athletes were more likely lo he symptomatic than low-LES athletes and that elevated cortisol level was positively correlated with symptomatology. To the extent that cortisol is a marker of exercise recovery in competitive athletes. our results suggest that chronic stress prolongs the recovery process, which may potentially widen a window of susceptibility for illness and injury among competitive athletes. PMID- 16250788 TI - Effects of reinforcing increases in active behavior versus decreases in sedentary behavior for obese children. AB - This experiment tested the effects of reinforcing obese children lo be more active or less sedentary in their choice of active versus sedentary behaviors. On days I and 5, there were no contingencies for sedentary or active behaviors. During days 2 through 4, children in the Activity group were reinforced for being more active, and they significantly increased their activity and decreased time spent on preferred sedentary activities. Children in the Sedentary group were reinforced for not engaging in preferred sedentary behaviors, and they significantly decreased time spent on these sedentary behaviors, with lime reallocated both to being more active and to substitution of lower preference sedentary behaviors. Children randomized to the Control group were reinforced for attendance and made choices among the alternatives as usual, allocating most or their time during all 5 days for their preferred sedentary behaviors. These laboratory results support the idea that activity can be increased by either reinforcing children for being more active or for reducing lime spent in sedentary activities. PMID- 16250789 TI - Somatoform symptoms in depressive and panic syndromes. AB - Somatoform symptoms are common features of psychological and psychosomatic disorders. This study addresses the question of whether somatoform symptoms differ in patients with panic syndromes. with depressive syndromes, or with somatization syndromes without depression or panic syndromes. We therefore investigated 135 inpatients o f a psychosomatic clinic and identified 64 patients for the depression group, 31 for the panic subgroup, and 18 for the somatization syndrome group. Neither the number of somatization symptoms nor the pattern of somatoform symptoms differed substantially among the 3 groups, except for higher frequencies of palpitations in the panic group and more abdominal pain symptoms in the depressive group. The 3 groups showed nearly identical frequency distributions of the individual somatoform symptoms. All 3 groups showed elevated hypochondriasis scores. In personality dimensions, depressive patients showed the lowest scores for extraversion. The improvements during inpatient treatment on the somatization variables, as well as general psychopathology, were also comparable. We favor the interpretation that the somatization syndrome is a fairly uniform syndrome whether or not it occurs alone or in combination with depressive syndromes or panic syndromes. PMID- 16250790 TI - Effects of gender-typed tasks and gender roles on cardiovascular reactivity. PMID- 16250791 TI - Health-related behaviors in the Republic of Karelia, Russia, and North Karelia, Finland. AB - Risk factors and health behaviors related to chronic diseases were studied in a population survey in Pitkaranta District in the Republic of Karelia, Russia, and in North Karelia, Finland, in Spring 1992 (Puska, Matilainen et al., 1993). The random sample of the population (25 to 64 years) was 1,000 in Pitkaranta and 2,000 in North Karelia. Among men there were more current smokers in Pitkaranta than in North Karelia (65% vs. 31%) whereas among women the respective rates were 11% and 16%. Self-reported alcohol consumption was higher in North Karelia. Leisure time physical activity was much less frequent in Pitkaranta both among men and women. Use of vegetables and berries was very infrequent in Pitkaranta. The differences in health behavior can at least partly explain the differences in risk factors and in mortality of chronic diseases. PMID- 16250792 TI - Anxiety, depression, and heart disease in women. AB - This is an extension of previous research that has reported on psychosocial risk factors in women participants in the Recurrent Coronary Prevention Project (RCPP). The RCPP women (N = 83) were under 65 years of age, non-diabetic, non smoking and had experienced a myocardial infarction (MI) at least 6 months prior to the study. Baseline data was available on 80 RCPP subjects. Seventy three non smoking, coronary disease-free women participants in the Stanford-Sunnyvale Health Improvement Project (SSHIP) served as a control-comparison group. Women with coronary heart disease had higher serum cholesterol than controls. There were no case-control differences in marital status, occupation, or number of children. RCPP women had Videotaped Structured Interview (VSI) Type A scores comparable to those of the SSHIP women, but had significantly higher VSI hostility scores (p < .01). In addition. the post-MI women were rated more anxious and depressed, and had more avoidance symptoms than controls ( p < .01). Additional analyses involved the 65 RCPP women located at 8.5-year follow-up. In these women, univariate predictors of coronary recurrence (N = 13) were body mass index (kg/m)2. Peel Index, low time urgency (VSI) and high anxiety ( p < .05). Employment status, marital status, and education were not associated with subsequent cardiac events. These exploratory analyses suggest that the relations between heart disease and hostility, anxiety, and depression in women deserve further investigation. PMID- 16250793 TI - Role of nicotine dependence in smoking relapse: results from a prospective study using population-based recruitment methodology. AB - Results of a prospective examination of factors influencing smoking relapse are reported for a sample of smokers (N = 522) obtained through population-based recruitment. All subjects were given a modified Fagerstrom Tolerance Questionnaire (modified FTQ), which is a 5-item self-report instrument characterizing smokers according to their degree of nicotine dependence. At 30 days post-cessation, those placing in the lowest quartile of the modified FTQ at baseline had the highest survival rate (79%) and those placing in the highest quartile at baseline had the lowest survival (non-relapse) rate (43%). The difference in survival among the groups was statistically significant (p < .001). At 12 months, those placing in the lowest quartile of baseline cigarette consumption had the highest survival rate (34%) and those placing in the highest quartile had the lowest survival rate (14%; p < .001). It seems increasingly clear that level of dependence, even as indexed by comparatively crude self report measures such as amount smoked is associated with the rate and pattern of smoking relapse. PMID- 16250794 TI - 1-year stability and prediction of cardiovascular functioning at rest and during laboratory stressors in youth with family histories of essential hypertension. AB - Blood pressure (BP). heart rate, cardiac index, and total peripheral resistance index were measured at rest and in response to postural change, forehead cold stimulation, and a video game challenge in a sample of 128 White and 155 African American normotensive youth with family histories of essential hypertension (EH). These measurements were readministered 1 year later (12.5 +/- 3.2 months). Moderate temporal stability was observed for all resting and absolute stress responses. Reliability estimates for reactivity change scores were lower, although some were within acceptable ranges. African-American youth exhibited greater BP and peripheral resistance index reactivity to forehead cold on both evaluations. After controlling for various anthropometric and demographic parameters and the pertinent previous year's resting cardiovascular (CV) parameter, mean video game systolic blood pressure (SBP) responses were predictive of resting SBP whereas absolute forehead cold and video game diastolic responses predicted resting diastolic blood pressure (DBP) 1 year later. Mean video game DBP responses were also predictive of resting peripheral resistance index after controlling for significant demographic and anthropometric measures. CV reactivity is discussed with regard to possible value in prediction of changes in resting BP and cardiac structure prior lo establishment of EH. PMID- 16250795 TI - Psychophysiological stress and EMG activity of the trapezius muscle. AB - Although it is generally assumed that mental stress induces muscular tension, the experimental data have, so far, been inconclusive. Likely explanations for these inconsistent findings are (a) too small subject samples in some experiments, (b) the use of only one type of stress stimulation, and (c) the lack of objective (physiological) measurements documenting the stress-inducing properties of the experimental treatment. Furthermore. the effect of mental stress and physical load separately, versus the combined influence of physical and mental load on muscular tension, has not been investigated earlier. Therefore, the aim o f the present experiment was lo examine the effects of mental stress as well as of physical load, separately and in combination, on perceived stress, physiological stress responses, and on muscular tension as reflected in electromyographical (EMG) activity of the trapezius muscle. Sixty two female subjects were individually exposed to mental arithmetic, the Stroop color word test (CWT), the cold pressor test, standardized test contractions (TCs), and the CWT combined with a TC. Compared to baseline, the stress session induced significant increases in systolic and diastolic blood pressure, heart rate, urinary catecholamines, salivary cortisol, and self-reported stress. Each of the two mental stress tests induced a significant increase in EMG activity. The CWT caused a rise in EMG activity also during the TC, which was significantly more pronounced than the increase induced by the CWT alone. Blood pressure responses and self-reported stress followed the same pattern as the EMG activity. The results are consistent with the assumption that psychological stress plays a role in musculoskeletal disorders by increasing muscular tension both in low-load work situations and in the absence of physical load. It is also indicated that the stress-induced increase in muscular tension is accentuated on top of a physical load. PMID- 16250796 TI - Exercise and dietary modification with women of non-English speaking background: a pilot study with Polish-Australian women. AB - Health promotion programs aimed at the general community often fail to reach Australians of non-English speaking background (NESB) because of language and cultural barriers. A 12-week minimal-intervention heart health program, designed for women of European background, was piloted with 43 women from a Polish social group, with a further 30 women serving as a comparison group. Assessments of the intervention group before and after the 12-week program indicated significant decreases in exercising heart rate and in resting blood pressure, which were not evident in the comparison group. Twelve-week follow-up data indicated that these gains had been well maintained. When the comparison group was invited to participate in a similar program, there were also significant improvements on these variables. This pilot study suggests that health promotion programs aimed at NESB Australians can be effective in modifying risk factors for cardiovascular disease if an effort is made lo address language and cultural barriers. PMID- 16250798 TI - Sensory effects of baroreceptor activation and perceived stress together predict long-term blood pressure elevations. AB - Activating the arterial baroreceptors in animals has been shown to blunt pain sensation and provide other forms of central nervous system inhibition. This study tested the hypothesis that, among human subjects, a tonic increase in blood pressure (BP) could be a learned response to environmental stressors among subjects in whom the baroreceptor inhibitory mechanism is active. In a sample of 96 healthy, normotensive men and women, amount of pain-reduction produced by baroreceptor stimulation predicted an increase in resting BP 20 months later: the increase was proportional to self-assessed daily life stress. Among the subjects reporting the greatest amount of stress, the pain inhibition effect accounted for more than 80% of the BP variance. These results support the hypothesis that the reduction in perceived stress produced by baroreceptor stimulation may reward learned increases in BP. PMID- 16250797 TI - Lipid lowering through work stress reduction. AB - Swedish civil servants, who reported considerable stress while working in larger government authorities, were randomized into either a work stress reduction intervention or a control group. Four work units constituted the intervention group (n = 94). and one unit formed the control group (n = 35). Participants in the two groups were of similar age and two thirds were women. Psychosocial, lifestyle, and physiological assessments were made before, during, and after the 8-month intervention period. A significant decrease in apolipoprotein B/apolipoprotein A I ratio (p < 0.05) occurred in the intervention group but not in the control group. No changes were seen in either group in smoking, eating. exercise, relative weight, or other lifestyle factors. which could explain the lipid profile changes. Stimulation from and autonomy over work increased in the intervention group (p < 0.01) hut remained the same in the control group. Results have implications for the efficacy of work stress interventions in decreasing coronary risk. PMID- 16250799 TI - Comparison of cardiac versus vascular reactors and ethnic groups in plasma epinephrine and norepinephrine responses to stress. AB - This study examined differences in plasma epinephrine (EPI) and norepinephrine (NOREPI) responses to stressors in 67 healthy African-American and Caucasian American men and women of the ages 18 to 49. Subjects were divided into three groups: (a) those who showed high blood pressure (BPI responses to stress associated with consistently high cardiac output (CO) increases with no substantial increases in total peripheral resistance (TPR), labeled cardiac reactors: (b) those with equally high BP increases associated with consistently higher TPR increases and lesser CO increases. labeled vascular reactors; (c) those who showed mixed hemodynamic responses or were low BP reactors. Ethnic and gender group differences in EPI and NOREPI responses were also examined. Cardiac reactors, vascular reactors and mixed + low reactors did not differ in EPI or NOREPI levels at baseline. During stressors, cardiac reactors showed greater increases in plasma EPI than vascular reactors or others during math, reaction time, and passive and active speech tasks: they also showed a weak trend toward greater NOREPI increases during these challenges as well. No differences were seen during the cold pressor: this stressor evoked the least change in EPI of all tasks, whereas the active speech elicited the greatest increases in both EPI and NOREPI of all tasks. Vascular reactors did not differ from mixed + low reactors in EPI or NOREPI reactivity, and men and women did not differ in EPI or NOREPI reactivity to any task. In contrast, Black subjects showed greater increases in NOREPI levels across all stressors compared to White subjects. These findings have implications for investigations of the role of sympathetic nervous system activity in the pathogenesis of hypertension in both African-American and Caucasian American populations. PMID- 16250800 TI - Psychophysiological effects of habitual caffeine consumption. AB - Caffeine is the most widely consumed pharmacologically active substance in the world, and a key issue concerning its possible implications for human health is whether it has persistent (i.e., chronic) physiological effects on habitual consumers. This study examined blood pressure, heart rate (HR), electromyogram (EMG), and skin conductance level (SCL) in 36 healthy men and women exposed to a pattern of moderate intake. A double-blind placebo-controlled crossover design with counterbalancing was used in which all subjects participated in four experimental conditions involving the ingestion of placebo or caffeine three times daily for 6 days followed by a seventh ("challenge") day of placebo or caffeine ingestion. Results confirmed that caffeine has significant pressor effects, and these were found to he additive to the pressor action of a laboratory stressor. Following habitual consumption of the drug. pressor effects were diminished (indicative of tolerance) but not eliminated. Effects of caffeine on other parameters were either modest (HR and EMG) or negligible (SCL). Considering (he near-universal use of caffeine. the persistent pressor effects observed in this study have important implications for clinical practice and public health. PMID- 16250801 TI - Association between smoking status and cardiovascular and cortisol stress responsivity in healthy young men. AB - Acutely, cigarette smoking stimulates increases in blood pressure (BP), heart rate (HR), and cortisol, but little evidence is available concerning the impact of habitual smoking status on cardiovascular stress responsivity. This relation was assessed in 86 healthy male firefighters, age 19 to 31. comprising 52 nonsmokers and 34 smokers. Measures of BP, HR, salivary free cortisol, breathing pattern, and self-reported stress and alertness were obtained while subjects performed nonverbal mental arithmetic and a socially evaluative speech task. Systolic and diastolic BP were higher at rest in nonsmokers than smokers, and a consistent difference in stress responsivity was also found. BP, HR, and cortisol responses to mental arithmetic were significantly smaller in smokers than nonsmokers, with mean changes in BP (adjusted for body weight) averaging 19.3/11.0 mmHg and 28.5/15.4 mmHg in smokers and nonsmokers, respectively. There were no effects of smoking status on task performance or subjective stress responses and no differences between groups in family health history, health related behaviors, o r psychological characteristics that might account for the reactivity difference. Possible explanations of the results are discussed, and methodological implications for cardiovascular stress reactivity studies are outlined. PMID- 16250804 TI - Modification of the Type A behavior pattern in post-myocardial infarction patients: a route to cardiac rehabilitation. PMID- 16250803 TI - Cardiovascular reactivity in cardiovascular disease: "once more unto the breach". AB - Discussed here are conceptual and methodologic issues that bear on the role of behaviorally evoked cardiovascular reactivity in cardiovascular disease. It is argued that recent criticisms concerning the validity of cardiovascular reactivity as a stable dimension of individual differences arise from inadequacies of measurement prevalent in prior literature. With standardization of test stimuli and application of psychometric principles lo protocol development, assessment of reactivity are found to be highly reliable and, in turn, to demonstrate the dispositional nature of this construct. Recent studies also document an underlying heterogeneity of hemodynamic reactions to stress, with distinct cardiac and vascular components. Because hemodynamic adjustments show some plasticity under differing task conditions, responses seen in particular contexts reflect influences of both an individual-specific response potential and response-eliciting properties of the stimulus. On the question of disease relevance, it is concluded that cardiovascular reactivity cannot yet be considered an established risk factor for either coronary heart disease or hypertension. However, the preponderance of existing clinical, experimental. and epidemiologic evidence is consistent with such an association and warrants further study in the context or population-based, prospective investigation. PMID- 16250806 TI - Epidemiology of headache in children and adolescents: evidence of high prevalence of migraine. PMID- 16250805 TI - Psychophysiological effects of cardiac rehabilitation in post-myocardial infarction patients. AB - Psychophysiological reactivity and cardiovascular functioning were assessed in 40 patients with coronary heart disease before and after taking part in either a Type A modification training program or a control condition. Treated patients showed an overall attenuated heart rate (HR) response and prolonged diastolic time to mental stress performance, during recovery and rest periods, as compared lo control patients. No other measure, including impedance-cardiography-derived measures, differentiated the two groups. The treatment group showed reduction from before lo after treatment in 24-hr minimum IIR level and decreased frequencies of premature ventricular contractions, whereas the control group showed an opposite trend. Results from an exercise test suggested the same direction for resting IIR and double-product levels. The two groups did not differ in maximal systolic and diastolic blood pressure or in maximal double product levels, despite a differential development, in favor of the treatment group, in maximal work capacity. PMID- 16250808 TI - Atopic dermatitis and stress: possible role of negative communication with significant others. AB - Clinical reports have related stressful social interactions to the course of atopic dermatitis (AD). Controlled behavioral observations of interaction patterns of AD patients and their significant others, however, were lacking. In two studies, adult AD patients and their mothers (N1 = 26) or partners (N2 = 67) engaged in a discussion of a mutual problem. Their verbal and nonverbal communication behavior was analyzed with a reliable standardized interactional coding system and compared to that of control dyads (N1 = 18, N2 = 28). Although AD patients and their significant others did not differ from controls in their self-reported satisfaction with the relationship, the behavioral observations showed less positive and more negative communication patterns compared to the control dyads. Negative interactions with significant others may prevent the solution of everyday problems and may add to the patients' stress level. PMID- 16250807 TI - Distress, denial, and low adherence to behavioral interventions predict faster disease progression in gay men infected with human immunodeficiency virus. AB - This study examined psychological prediction of 2-year disease progression in gay men after finding out their human immunodeficiency virus (HIV) serostatus. Psychological and immune status of asymptomatic gay men who did not know their HIV serostatus was monitored during the 5 weeks before and after serostatus notification. The men were randomly assigned to an exercise. cognitive-behavioral stress-management intervention, or control group. At 2-year follow-up for the 23 men who turned out to be seropositive. 9 had developed symptoms, including 5 with acquired immune deficiency syndrome--4 of whom died. Distress at diagnosis, denial (5 weeks post-diagnosis minus pre-diagnosis). and low adherence during interventions were significant predictors of 2-year disease progression. Denial and adherence remained significant predictors of disease progression even after controlling for CD4 number at entry. Furthermore. change in denial was significantly correlated with immune status 1 year later; l-year immune status was significantly correlated with 2-year disease progression. The present study therefore demonstrates significant relations between psychological variables on the one hand and both immune measures and HIV-1 disease progression on the other. We conclude that distress, denial, and low protocol compliance predict subsequent disease progression. PMID- 16250809 TI - Parental prompting of smoking among adolescents in Tijuana, Mexico. AB - Latino adolescents in the United States and Mexico may have higher rates of tobacco experimentation than other ethnic groups, possibly due to cultural factors and parental influences. This study examined three parental behaviors that may prompt smoking in children in Grades 3, 5, 7, and 9 in Tijuana. Mexico. Surveys were administered to 758 students in randomly selected classes in randomly selected schools in Tijuana. The most frequent prompt was the smoking parent asking the child to buy cigarettes (about two thirds), whereas about 60% asked the child to light the parent's cigarette, and about 20% of smoking parents asked the child to place the cigarette in his or her mouth to light it. Rates of prompting were very low among third graders but increased by Grade 5. Mother's smoking was associated with higher rates of all three types of parental prompting. PMID- 16250811 TI - Differential effects of active and passive laboratory stressors on immune function in healthy men. AB - The immunomodulatory effects of acute laboratory stressors were examined by comparing active and passive stressors in a between-subjects design. Healthy male volunteers (N = 67) were recruited and randomly assigned to an active, passive, or no stressor condition. Subjects were exposed to either the Stroop and mental arithmetic tasks (active). two surgery films (passive). or two nature films (no stress). Cardiovascular reactivity, plasma catecholamines and cortisol. and self reported distress were measured pre-task and post-task. Lymphocyte proliferation to concanavalin A (Con A). pokeweed mitogen (PWM), and phytohemagglutinin was assessed a1 baseline, after the first task, after the second task, and 30 min later. Lymphocyte proliferation lo Con A and PWM was significantly reduced in response to the stressors. Different response patterns emerged, depending on the type of stressor and the mitogen used. Changes in lymphocyte proliferation were significantly associated with cardiovascular reactivity during the tasks. Results are discussed in terms of potency of the stressors and mechanisms underlying passive versus active laboratory tasks. Implications for future research are addressed. PMID- 16250810 TI - Resting parasympathetic status and cardiovascular response to orthostatic and behavioral challenges in type I insulin-dependent diabetes mellitus. PMID- 16250812 TI - Disasters--how can telemedicine help? PMID- 16250814 TI - Telethinking with Nina M. Antoniotti, R.N., M.B.A., Ph.D. Interview by Vicki Glaser. PMID- 16250815 TI - The Missouri Telehealth Network. PMID- 16250816 TI - Telemedicine and the academic health center: the University of Michigan health system model. AB - The changing health care environment is generating a number of challenges for Academic Health Centers' (AHC) ability to pursue their traditional tripartite mission of medical care, education, and research. A number of strategies have been suggested to aid the AHC to respond, among them telemedicine. In this paper, telemedicine is examined for its potential to assist in meeting financial, cost, and quality challenges. In particular, the model developed at the University of Michigan Health System for implementing telemedicine within the AHC is presented together with lessons learned. This model, based largely on intramural rather than extramural funding, is offered as a basic strategy to be considered by AHCs facing these challenges. PMID- 16250818 TI - A retrospective evaluation of TeleMental Healthcare services for remote military populations. AB - The objective of this study was to compare specific treatment and outcome variables between mental health care via videoconferencing to care provided in person. The study was a retrospective record review of service members and their adult civilian family members seen at two remotely located military bases. One group was seen via video conferencing (telemental health care [TMHC]) while a second group was seen face-to-face care (FTFC) and served as a control group. Chi square tests were used to test for significance associations between therapy format and secondary variables. The Global Assessment of Functioning was unexpectedly and significantly more improved for the TMHC group than the FTFC care group. Mean change in Global Assessment of Functioning for FTFC (8.4) was significantly less than mean change for TMHC (15.3). There were no significant differences between the groups in the number of laboratories or studies ordered, self-help recommendations made, selected mental status elements, or number of patients prescribed two or more psychotropic medications. The rate of full compliance with the medication plan and follow-up appointments was significantly better for TMHC. Providers using TMHC told more patients to return for follow-up appointments in 30 days or less. Improved compliance, the unique interpersonal processes of care via TMH, and slightly shorter times to next follow-up appointment were the chief contributors. Alterations in the process of communication may have implications in the business, political, and military sectors. PMID- 16250817 TI - Performance model for telehealth use in home health agencies. AB - Increasingly, home health agencies (HHAs) are considering the value of implementing telehealth technology. However, questions arise concerning how to manage and use this technology to benefit patients, nurses, and the agency. Performance models will be beneficial to managers and decision makers in the home health field by providing quantitative information for present and future planning of staff and technology usage in the HHA. This paper presents a model that predicts the average daily census of the HHA as a function of statistically identified parameters. Average daily census was chosen as the outcome variable because it is a proxy measure of an agency's capacity. The model suggests that including a telehealth system in the HHA increases average daily census by 40% 90% depending on the number of nurse full-time equivalent(s) (FTEs) and amount of travel hours per month. The use of a home telecare system enhances HHA performance. PMID- 16250819 TI - A feasibility study of a personalized, internet-based compliance system for chronic disease management. AB - This paper describes a feasibility study on an Internet-based compliance system to provide personalized care for patients suffering from chronic diseases. Relevant medical trials from three different European countries provided preliminary evidence of the feasibility of the system and its efficacy in helping patients to manage their diseases at home. The study discusses further improvements not only for the C-Monitor system, but also for other Internet-based health-care services. PMID- 16250820 TI - Telepsychiatry program for rural victims of domestic violence. AB - Domestic violence is a significant public health problem and is correlated with serious mental and physical disorders. Victims' fear and isolation seriously limit access to psychiatric evaluation and treatment. Telemedicine provides a means to overcome these obstacles. This article describes a telemedicine program that provides psychiatric screening, evaluation, treatment, and referral for ongoing care to clients of a rural women's crisis center. Psychiatric evaluation and treatment were provided to a rural women's shelter program using telepsychiatry. The shelter program had difficulty accessing traditional mental health service. All new clients entering the program were screened for mental health problems. Those requiring further evaluation received a physical examination with medical history and initial psychological interview on site, followed by psychiatric evaluation by videoconference. Appropriate treatment was initiated, and referral for ongoing psychiatric care through the local community mental health clinic was arranged. Of the 38 women referred for mental health services by clinic staff, 35 completed a psychiatric evaluation using telepsychiatry and 31 entered treatment. The most commonly identified disorders were anxiety and major affective disorders, followed by substance use disorders. Telepsychiatry can provide rapid crisis intervention and effective mental health services to victims of domestic violence in a rural setting. PMID- 16250821 TI - Rural jail telepsychiatry: a pilot feasibility study. AB - New York State has a large rural population, and many of the jails in rural areas have minimal or no psychiatric services available on site. Cost of transport to off-site psychiatric services and the safety issues related to moving inmates from a secure building may limit inmate access to appropriate psychiatric services. This feasibility study describes a project that provided telepsychiatric consultation to increase access to psychiatric treatment in an underserved rural jail in upstate New York. Subjects were consenting jail inmates who requested or were found to be in need of psychiatric care. The project provided interactive two-way audio-video communication between the psychiatrist located in an urban university medical center and subjects who were incarcerated 182 miles away. During the project period, 15 inmates were assessed and treated in 37 consultations. Subjects were predominantly young white males with anxiety, mood, and substance use disorders. Services were readily accepted by inmates and staff. Telepsychiatric examination and treatment appears to be a feasible method to increase access to mental health care in rural jails. Future advocacy for increased mental health services in rural areas in criminal justice setting is likely to depend on further evidence of favorable cost benefit. PMID- 16250822 TI - Web-based analysis of nasal sound spectra. AB - The spectral analysis of the nasal sound is an indicator of the nasal airflow pattern. We investigated a new technique for nasal sound analysis via Internet. This study includes 27 patients and 22 healthy people. Patients were treated by septoplasty operation for septal deviation. Postoperation 10(th) day, this technique was applied to follow nasal airflow course. The patients recorded the nasal sound by microphone into the computer as a .wav file and sent us via internet, all those records were evaluated by us. The results were sent back to themselves. The 11 patients who had nasal obstruction symptoms (group A) were called to the hospital to check. In the nasal sound analyses e-mails of those patients, the sound intensity was at high frequencies (2-4 kHz, 4-6 kHz) above 30 dB, but low (500-1000 Hz) and medium frequencies (1-2 kHz), are below then 10 dB. In the patients without nasal obstruction symptom (group B), the sound intensity was at high frequencies below 10 dB, but low and medium frequencies are above 20 dB. There was a statistically significant difference in sound intensity between group A and group B. In the endoscopical examination of those obstructions, which decreases the nasal airway, crusting formation in the nasal cavity was found. Web based nasal sound analysis is an important method to follow the postoperative course and the nasal airflow evaluation. The new method will save time and money, avoiding a return visit to the hospital unnecessarily. PMID- 16250823 TI - A synchronous communication experiment within an online distance learning program: a case study. AB - Student-teacher and student-student interactions in purely asynchronous distance learning courses are much lacking compared to similar interactions found in face to-face teaching, causing learners to experience feelings of isolation, thus reducing motivation and increasing dropout rates. We used PalTalk, an Internet text and audio chat client from AVM Software, Inc. (New York, NY), to offer our students live virtual classroom sessions within a unit of our online distance learning M.Sc. program in Healthcare Informatics. On-demand replays of audio excerpts from the sessions were also provided to accommodate absenteeism and for student review. Five students completed an evaluation questionnaire. Our results highlighted the potential merits of using synchronous conferencing to assist in fostering a sense of belonging to one supportive learning community among distance learners and improve educational outcomes. Students were very positive toward the real-time human interaction and voted for a 95/5 (asynchronous/synchronous percentages) blended delivery approach for a typical unit in our program. They also praised PalTalk's voice quality and ease of use. This paper presents educational and technological perspectives about this experiment in the form of a state-of the- art review, without intending to be statistically rigorous. However, robust research evidence is still required to convince educators fully about the benefits of synchronous communication tools and help them decide on the most suitable solutions for their particular circumstances. PMID- 16250824 TI - Validation of rule-based inference of selected independent activities of daily living. AB - This paper explores the validity of a rule-based inference method of selected independent activities of daily living (ADLs). An inexpensive ADL monitoring system was installed in the community for 37 days to monitor a middle-aged, healthy individual living alone. The subject was given a personal digital assistant (PDA), running custom activity diary software, and asked to record activities in real-time. Rule-based activity inference algorithms were refined on data from 17 days, and data from the remaining 20 days were used for validation. The chisquare statistic was computed for 2 x 2 contingency tables comparing activities detected by the algorithms to user-logged activities. The phi (r()) and Cohen's kappa (kappa) coefficients were computed as measures of correlation. After correcting for subject noncompliance in logging activities, the kappa correlation between the meal detection algorithm and the PDA record was 0.84, with 91% sensitivity, and 100% specificity. Similarly, the kappa correlation between the shower detection algorithm and the PDA record is 0.69, with 67% sensitivity and 100% specificity. The detection algorithms and the sensory data did not miss any main meals or showering activities recorded on the PDA. The results suggest that rule-based algorithms can successfully detect meal preparation and showering activities using simple low-cost detectors. The sensors and detection algorithms reported events not recorded by the occupant on the PDA attributed to reporting noncompliance. Overall, the PDA activity journal was a compromise between paper diaries, which are more time consuming to keep, and may result in higher noncompliance errors, and video recording, which is considered intrusive. PMID- 16250825 TI - Critical success factors for strategic telemedicine planning in New Zealand. AB - This research reviewed the health information systems (HIS) strategy of the New Zealand government and highlighted different gaps in the strategy, as raised by the different stakeholders involved in this strategy. To address such gaps, the government provided different critical success factors (CSFs) for the successful implementation of the national HIS strategy. The research assessed the strategic importance of telemedicine by highlighting its strengths, weaknesses, opportunities, and threats (SWOT) in health-care organizations. The research utilized the portrayed HIS strategy and CSFs to depict a strategy for telemedicine integration in health-care organizations in New Zealand taking into consideration its SWOT. The developed CSFs are of strategic importance to health care professionals, researchers, and policymakers interested in integrating telemedicine in health-care delivery at the national level in New Zealand and elsewhere. PMID- 16250826 TI - Can subspecialty cancer consultations be delivered to communities using modern technology?--A pilot study. AB - The objective of this project was to evaluate patient and physician acceptance of subspecialty oncologic teleconsultation for distant communities. Many newly diagnosed cancer patients have to travel several hours and long distances to attend specialty medical oncology consultations at our regional cancer center in Victoria, BC. Difficulties in recruiting of oncologists in Vancouver Island have prompted the search for other means to deliver subspecialty consultation closer to home. Teleconsultation seemed a possible model. Hence, 30 sequential patients with gastrointestinal (GI) malignancy referred from the Central Island region were seen after an informed consent via videoconferencing and 30 sequential patients were seen face to face in Victoria by one oncologist. Patients and the oncologist filled out a satisfaction questionnaire. The age, sex, proportion of patients who subsequently received chemotherapy, and the number of other co morbid conditions were similar in both groups. No difference was observed in patient satisfaction whether patients were seen via videoconference or in person. However, the oncologist felt the video did not go as well as face-to-face consultation. Patients were very satisfied with teleconsultation, and it saved them hours of travel. PMID- 16250827 TI - A Latin American telemedicine social perspective from a Colombian telemedicine center initiative. AB - This paper describes a collaborative initiative for developing a telemedicine model on for Colombia and Latin America from a social perspective. This was led by the Colombian Telemedicine Center (CTMC) with the participation of teams from Mexico, Costa Rica, Panama, and Argentina, in addition to on-going interaction with academic institutions such as Javeriana University, Icesi University, and Murcia University. PMID- 16250828 TI - T-cell receptor-like antibodies: novel reagents for clinical cancer immunology and immunotherapy. AB - Major histocompatibility complex class I molecules play a central role in the immune response against a variety of cells that have undergone malignant transformation by shaping the T-cell repertoire and presenting peptide antigens from endogeneous antigens to CD8+ cytotoxic T-cells. Diseased tumor or virus infected cells are present on class I major histocompatibility complex molecule peptides that are derived from tumor-associated antigens or viral-derived proteins. Due to their unique specificity, such major histocompatibility complex peptide complexes are a desirable target for novel approaches in immunotherapy. Targeted delivery of toxins or other cytotoxic drugs to cells which express specific major histocompatibility complex-peptide complexes that are involved in the immune response against cancer or viral infections would allow for a specific immunotherapeutic treatment of these diseases. It has recently been demonstrated that antibodies with the antigen-specific, major histocompatibility complex restricted specificity of T-cells can be generated by taking advantage of the selection power of phage display technology. In addition to their tumor targeting capabilities, antibodies that mimic the fine specificity of T-cell receptors can serve as valuable research reagents that enable study of human class I peptide major histocompatibility complex ligand presentation, as well as T-cell receptor peptide-major histocompatibility complex interactions. T-cell receptor-like antibody molecules may prove to be useful tools for studying major histocompatibility complex class I antigen presentation in health and disease as well as for therapeutic purposes in cancer, infectious diseases and autoimmune disorders. PMID- 16250831 TI - Pharmacological properties of indazole derivatives: recent developments. AB - The chemistry of indazole and its N-oxide derivatives is very well-known. Indazole derivatives were extensively studied as bioactive compounds, such as anti-aggregatory and vasorelaxant activity by NO release and increase of cGMP levels and anticancer effects, antimicrobial and antiparasitic properties, among others. Recently, the research and development in the medicinal chemistry of these systems have produced compounds with contraceptive activities for men, for the treatment of osteoporosis, inflammatory disorders and neurodegenerative diseases. On the other hand, indazole N-oxide derivatives were poorly studied as bioactive compounds, but recently compounds with antiparasitic properties were produced. In this presentation, recent developments in the chemistry and medicinal chemistry of indazole and its N-oxide derivatives will be reviewed. PMID- 16250832 TI - The structure-inhibitory activity relationships study in a series of cyclooxygenase-2 inhibitors: a combined electronic-topological and neural networks approach. AB - Structure-activity relationships study was performed for a few series of cyclooxygenase-2 (COX-2) inhibitors by using the Electronic-Topological Method combined with Neural Networks (ETM-NN). Specific molecular fragments were found for active compounds ('activity features') from both series by the ETM application. After this, a system of prognosis was developed as the result of training Kohonen's self-organizing maps (SOM) by the fragments. From the detailed analysis of all compounds under study, requirements necessary for a compound to be COX-2 inhibitor were formulated. The analysis showed that any requirements violation for a molecule resulted in a considerable decrease or even complete loss of its activity. The found activity features identified correctly different marketed drugs and new compounds that had passed pre-clinical and clinical trials; this fact confirms the workability of the system developed for the COX-2 inhibitory activity prediction. PMID- 16250833 TI - Advances in prodrug design. AB - The background of prodrug design is presented herein as the basis for introducing new and advanced latent systems, taking into account mainly the versatility of polymers and other macromolecules as carriers. PDEPT (Polymer-Directed Enzyme Prodrug Therapy); PELT (Polymer-Enzyme Liposome Therapy); CDS (Chemical Delivery System); ADEPT(Antibody-Directed Enzyme Prodrug Therapy); GDEPT/VDEPT (Gene Directed Enzyme Prodrug Therapy/Virus-Directed Enzyme Prodrug Therapy); ODDS (Osteotropic Drug Delivery System) and LEAPT (Lectin-directed enzyme-activated prodrug therapy) are briefly described and some examples are given. PMID- 16250834 TI - New anti-Alzheimer drugs from biodiversity: the role of the natural acetylcholinesterase inhibitors. AB - Alzheimer's disease (AD) is a progressive neurodegenerative pathology with severe economic and social impact. There is currently no cure, although cholinesterase inhibitors provide effective temporary relief of symptoms in some patients. Nowadays, drug research and development are based on the cholinergic hypothesis that supports the cognition improvement by regulation of the synthesis and release of acetylcholine in the brain. There are only four commercial medicines approved for treatment of AD, and natural products have played an important alternative role in the research for new acetylcholinesterase inhibitors, as exemplified through the discovery of galantamine. This profile conducts us to give in this paper an overview relating the several classes of natural products with anti-cholinesterasic activity as potential templates to the design of new selective and powerful anti-Alzheimer drugs. PMID- 16250835 TI - Pharmacophore identification for sigma-1 (sigma1) receptor binding: application of the "deconstruction-reconstruction-elaboration" approach. AB - At least two different types of sigma (sigma1 and sigma2) receptors have been identified. A structural feature common to high-affinity (Ki <10 nM) sigma1 ligands is: C-N(R)-X-Ph; both C and Ph are associated with regions of bulk tolerance. Numerous other ligands bind, but typically do so with lower affinity. PMID- 16250836 TI - Cannabinoids and cancer. AB - Marijuana has been used in medicine for millennia, but it was not until 1964 that delta9-tetrahydrocannabinol (delta9-THC), its major psychoactive component, was isolated in pure form and its structure was elucidated. Shortly thereafter it was synthesized and became readily available. However, it took another decade until the first report on its antineoplastic activity appeared. In 1975, Munson discovered that cannabinoids suppress Lewis lung carcinoma cell growth. The mechanism of this action was shown to be inhibition of DNA synthesis. Antiproliferative action on some other cancer cells was also found. In spite of the promising results from these early studies, further investigations in this area were not reported until a few years ago, when almost simultaneously two groups initiated research on the antiproliferative effects of cannabinoids on cancer cells: Di Marzo's group found that cannabinoids inhibit breast cancer cell proliferation, and Guzman's group found that cannabinoids inhibit the growth of C6 glioma cell. Other groups also started work in this field, and today, a wide array of cancer cell lines that are affected is known, and some mechanisms involved have been elucidated. PMID- 16250837 TI - Corticotropin-releasing factor binding protein--a ligand trap? AB - The actions of the neuropeptide corticotropin-releasing factor (CRF) are modulated by a CRF binding protein (CRFBP). In view of the memory-enhancing effects of CRF, the release of endogenous CRF from CRFBP by CRFBP inhibitors has been suggested as a therapeutical strategy for the treatment of cognitive deficits. This mini-review will summarize recent advances in the field with a focus on the pharmaceutical potential of CRFBP inhibitors. PMID- 16250839 TI - Phosphodiesterase inhibitors for cognitive enhancement. AB - An effective treatment for age-related cognitive deficits remains an unmet medical need. Currently available drugs for the symptomatic treatment of Alzheimer's disease or other dementias have limited efficacy. This may be due to their action at only one of the many neurotransmitter systems involved in the complex mechanisms that underlie cognition. An alternative approach would be to target second messenger systems that are utilized by multiple neurotransmitters. Cyclic adenosine monophosphate (cAMP) is a second messenger that plays a key role in biochemical processes that regulate the cognitive process of memory consolidation. Prolongation of cAMP signals can be accomplished by inhibiting phosphodiesterases (PDEs). Eleven PDE families, comprised of more than 50 distinct members, are currently known. This review summarizes the evidence demonstrating that rolipram, a selective inhibitor of cAMP-selective PDE4 enzymes, has positive effects on learning and memory in animal models. These data provide support for the general approach of second messenger modulation as a potential therapy for cognitive dysfunction, and specifically suggest that PDE4 inhibitors may have utility for improving the symptoms of cognitive decline associated with neurodegenerative and psychiatric diseases. PMID- 16250840 TI - The dualistic nature of immune modulation in Alzheimer's disease: lessons from the transgenic models. AB - There is extensive evidence that changes in immune system activation accompany the pathological changes of Alzheimer's disease (AD), but a mechanistic understanding of how the immune system actually participates in disease pathogenesis is still largely lacking. Because of the complexity of the immunological response, and the difficulty in identifying the key molecular players that underlie any given immunological response, expanding our understanding of the immunological response in AD beyond its descriptive stages has not been a straightforward exercise. The development of transgenic animals that form deposits of Abeta peptide in their brains has provided an unexpected dividend to those interested in the immunological response characterizing AD. Several of these transgenic models develop structures greatly resembling neuritic plaques, a hallmark feature of AD brain that is also a focal point of the immunological response occurring in AD. Genetic and pharmacological manipulation of these Abeta-depositing transgenic mice is providing some intriguing and unexpected insights into the role of innate and adaptive immune mechanisms in the pathogenesis of AD. This review will discuss immunological perspectives that have arisen from research using Abeta-depositing transgenic mice, and place these perspectives in the context of epidemiological and genetic studies that have previously suggested a role for the immune system in AD. The emerging story affirms the likely role of innate and adaptive immune mechanisms in the pathogenesis of AD, but provides a cautionary note as to the difficulties that are likely to face potential immunomodulatory therapies due to the dualistic beneficial and detrimental roles that immune mechanisms appear to play in AD. PMID- 16250841 TI - Muscarinic receptor agonists and antagonists in the treatment of Alzheimer's disease. AB - One of the consistent findings in the brains of Alzheimer's Disease (AD) patients is loss of cholinergic function. The cholinergic approach to treatment of AD involves counteracting this loss in cholinergic activity by pharmacological intervention to increase cholinergic transmission. The cognitive effects of acetylcholine are mediated via the muscarinic M1 receptor. Direct stimulation of this receptor using muscarinic M1 agonists improves cognition in animal models and improves performance in cognitive tests in Alzheimer's patients. Alternatively, antagonists of central presynaptic M2 receptors improves cognition by increasing the central release of acetylcholine. Both approaches require high selectivity for one muscarinic receptor sub-type both for efficacy and to avoid cholinergic side effects. In this review summarizes recent progress in the identification and characterization of selective muscarinic receptor ligands for the treatment of Alzheimer's disease. PMID- 16250842 TI - Gamma-secretase as a therapeutic target for the treatment of Alzheimer's disease. AB - An effective, disease-modifying treatment of Alzheimer's disease (AD) remains one of the most significant unmet needs in modern medicine. As a result of the extensive research in the area, the mechanisms underlying the disease are now much better understood than at any time before. A significant amount of evidence points to the central role of beta-amyloid (Abeta) peptide-mediated toxicity in the disease etiology and strategies to remove this species from the central nervous system (CNS) have been actively pursued. The enzyme responsible for the final step in Abeta synthesis, gamma-secretase, has emerged as an attractive drug target and intensive research has transformed this enzyme from shadowy beginnings into a well characterised member of a new family of intramembrane-cleaving aspartyl proteases. Many inhibitors across diverse structural classes have been discovered and have demonstrated a lowering of central Abeta levels in preclinical models of AD. It has also become increasingly evident more recently that gamma-secretase also mediates a range of cleavages of alternative transmembrane peptides most notably the Notch receptor and the functional consequences of this activity have attracted much attention. The ultimate therapeutic benefit of gamma-secretase inhibitors and the effect of alternative, mechanism-based activities can only be judged when clinical data is forthcoming. In this review we describe the literature regarding the discovery of the nature of gamma-secretase, the development of small molecule inhibitors and their in vivo profiles. PMID- 16250844 TI - RNA interference-based gene silencing in mice: the development of a novel therapeutical strategy. AB - RNAi (RNA interference) was originally detected in Caenorhabditis elegans as biological response to exogenous double-stranded RNA (dsRNA), which induces very effective sequence-specific silencing of gene expression. Further investigations revealed that RNAi can occur in many eukaryotic species. Increasing understanding of the biochemical components of RNAi indicates the existence of a conserved machinery for dsRNA-induced gene silencing that acts in two steps. In the first step, an RNase III family nuclease called Dicer processes the dsRNA to small interfering RNAs (siRNAs) 21-23 nt in length. These siRNAs enter a multimeric nuclease complex that identifies target mRNAs through their homology to siRNAs and induce destruction of the corresponding mRNAs. Since RNAi has become an excellent strategy for gene silencing, it is tempting to apply this technology to 'knock-down' gene expression in living animals. The generation of transgenic mice from embryonic stem cells expressing small hairpin RNAs (shRNAs) has provided evidence for in vivo application of RNAi. Furthermore, different experimental strategies have been developed to analyze the influence of chemically synthesized siRNAs and of vector-based shRNAs on the expression of different transgenes and endogenous genes in vivo. Recent studies describe the in vivo delivery of siRNAs to inhibit transgene expression in certain organs of adult mice, predominately murine liver. Strategies for the inhibition of cellular proliferation by systemic treatment of tumor-bearing animals with siRNAs are beginning to emerge. They are of utmost interest for systemic diseases such as cancer. In addition, several groups have shown that RNAi can also be used to block the infectivity or suppress the replication of different RNA viruses relevant to human diseases including human immunodeficiency virus-1 (HIV-1) and hepatitis C virus (HCV). In summary, multiple lines of evidence indicate that RNAi seems to become a powerful tool for the fight against undesirable gene expression in human diseases. PMID- 16250843 TI - Progress in the discovery of BACE inhibitors. AB - Dementia caused by Alzheimer's disease is a large medical burden on society in the developed world. Current treatments are largely symptomatic, and there is an urgent need for therapies which can interrupt or reverse the progression of disease. A number of strategies for intervention are being actively pursued; among the most promising is the inhibition of beta-secretase, or BACE. BACE is the enzyme responsible for N-terminal cleavage of the Alzheimer's precursor protein leading to the production of the beta-amyloid peptide. This cascade ultimately leads to the formation of amyloid plaques, one of the hallmark lesions of the disease. It is expected that inhibitors of BACE may therefore serve as an effective disease-modifing therapy for the treatment of AD. This concept has received significant attention by both academics and the pharmaceutical industry. This review focuses on a discussion of the reported structure-activity relationships for inhibitors of this important therapeutic target. PMID- 16250845 TI - Participation of the cannabinoid system in the regulation of emotional-like behaviour. AB - The endocannabinoid system has been involved in the control of several neurophysiological and behavioural responses. Indeed, recent studies have suggested that the cannabinoid system could represent an important substrate for the control of emotional behaviour, and further research would probably help to identify new promising therapeutic targets. This paper reviews the results obtained in different animal models used to investigate emotional states after the manipulation of the endocannabinoid system. Cannabinoid compounds can induce anxiogenic- and anxiolytic-like responses in rodents depending on the experimental conditions. Studies using knockout mice lacking the CB1 cannabinoid receptors have shown the participation of this receptor in several behavioural responses including anxiety- and depressive-like states. Furthermore, the endocannabinoid system regulates the hypothalamic-pituitary adrenal axis, which is involved in providing an appropriate response to stressful situations. Recent studies have also demonstrated that the endocannabinoids can function as retrograde messengers, modulating the release of different neurotransmitter, including opioids, GABA and cholecystokinin that have been classically involved in the control of anxiety-like responses. All this recent information has further clarified the role played by the endogenous cannabinoid system in the control of emotional behaviour and provides data to support a new possible therapeutic use of cannabinoid compounds. PMID- 16250846 TI - Pharmacological intervention of cyclooxygenase-2 and 5-lipoxygenase pathways. Impact on inflammation and cancer. AB - Eicosanoids are potent biologically active arachidonic acid-derived lipid mediators that are intimately involved in inflammation and cancer. Cyclooxygenase (COX), the key enzyme in prostaglandin (PG) biosynthesis, controls one of the major pathways of arachidonic acid metabolism and is the main target for non steroidal anti-inflammatory drugs (NSAIDs). COX exists in two distinct isoforms, COX-1 and COX-2, the latter being primarily involved in inflammation and cell proliferation. For this reason, in recent years, selective COX-2 inhibitors, that achieve the same anti-inflammatory efficacy as traditional NSAIDs but minimize the risk of unwanted side-effects, have been developed. On the other hand, emerging information has appreciated the role of other arachidonic acid metabolic pathway (the 5-lipoxygenase (5-LO) pathway) in producing and maintaining inflammation. Moreover, it is now being perceived that COX-2 and 5-LO have converging functions not only in inflammation but also in cell proliferation and neo-angiogenesis. In this regard, there is evidence that COX-2 and 5-LO are co expressed and up-regulated in a number of inflammatory and neoplastic disorders, and that COX-2 as well as 5-LO inhibitors have beneficial effects in inflammatory diseases and are being investigated as potential anticancer drugs. This review provides an overview and an update of the progress achieved in the knowledge of COX-2 and 5-LO pathways and their involvement in inflammation and cancer. It also proposes a model of integrated pharmacological intervention on these pathways and reviews the information available regarding the use of the novel dual COX-2/5-LO inhibitors that block both pathways equally well. PMID- 16250847 TI - FLT3 and acute myelogenous leukemia: biology, clinical significance and therapeutic applications. AB - Acute myelogenous leukemia (AML) is a difficult disease to treat, and better treatments are needed. Molecular targeted therapy represents a novel therapeutic approach. The FLT3 tyrosine kinase receptor is mutated in approximately one fourth to one-third of patients with AML. Normally, binding of FLT3 ligand to the FLT3 receptor leads to phosphorylation of tyrosine residues and activation of the receptor. This in turn leads to induction of intracellular signaling pathways essential to regulation of cell proliferation and apoptosis. Two classes of FLT3 activating mutations have been identified in AML patients: internal tandem duplications (ITDS) and point mutations in the activating loop of the kinase domain. Both mutations result in constitutive FLT3 tyrosine kinase activity and lead to transformation of hematopoietic cell lines in vivo and in vitro. FLT3 ITDs are also an independent poor prognostic factor for overall survival and disease free survival in patients with AML. Therefore, targeting FLT3 mutations represents a potential therapeutic target for AML. This review will discuss the biology and clinical significance of FLT3 and FLT3 mutations in cell growth and signaling. In addition, I will discuss some of the novel FLT3 inhibitors which are entering clinical trials for AML. PMID- 16250848 TI - Large artery stiffness and antihypertensive agents. AB - PURPOSE OF REVIEW: Since in hypertensive populations, concentration on peripheral blood pressure only does not achieve 100% of blood pressure-attributable risk reduction, taking into consideration other hemodynamic parameters than peripheral blood pressure could perhaps improve cardiovascular prevention. The main purpose of this review is to analyse the scientific data in favour of considering arterial stiffness parameters as interesting intermediate cardiovascular endpoints in order to optimise risk assessment and risk reduction strategies. SUMMARY: Aortic pulse wave velocity (PWV), a marker of aortic stiffness, has been shown to be a strong independent predictor of cardiovascular morbid events, cardiovascular and all-cause mortality in numerous studies in different populations. Furthermore, it has been shown in a therapeutic trial that the lack of aortic PWV attenuation despite significant drug-induced reduction in mean blood pressure was a significant predictor of cardiovascular death in subjects with end-stage renal disease. In essential hypertension, the Reason Study has shown that, despite a similar decrease in peripheral diastolic blood pressure, different effects on central hemodynamic parameters were observed between blockade of the renin-angiotensin system and atenolol. Novel therapeutic approaches available to reduce the increase of pulse pressure and arterial stiffness with age involve converting enzyme inhibitors in association with diuretic compounds; nitrate derivatives; agents acting on collagen cross-linking; and finally spironolactone and vasopeptidase inhibitors. CONCLUSION: These results support the hypothesis that measurement of aortic PWV could then help, not only in risk assessment strategies but also in risk reduction strategies by monitoring arterial stiffness under different pharmacological regimens. PMID- 16250850 TI - PET imaging in clinical drug abuse research. AB - Over the last two decades, SPECT (single photon emission computed tomography) and especially PET (positron emission tomography) have proven increasingly effective imaging modalities in the study of human psychopharmacology. Abusing populations can be studied at multiple times after abstinence begins, to give information about neurochemical and physiological adaptations of the brain during recovery from addiction. Individual human subjects can be studied using multiple positron labeled radiotracers, so as to probe more than one facet of brain function. PET and SPECT have been used to help our understanding of many aspects of the pharmacokinetics and pharmacodynamics of abused drugs, and have made valuable contributions in terms of drug mechanisms, drug interactions (e.g. cocaine and alcohol) and drug toxicities. They have also been employed to study the acute effects of drugs on populations of active drug abusers and of normal controls, and to evaluate the neurochemical consequences of candidate therapies for drug abuse. A particularly productive strategy has been the use of PET in conjunction with neuropsychological testing of subjects, to allow correlation of imaging data with uniquely human aspects of the effects of drugs, such as euphoria and craving. PMID- 16250851 TI - Fluorine-18-labelled fluoropyridines: advances in radiopharmaceutical design. AB - Positron Emission Tomography is a high-resolution, sensitive, functional imaging technique, which can efficiently give access to the distribution, pharmacokinetics and -dynamics of a drug in vivo and which can therefore advantageously play a key-role in both drug discovery and development. This molecular imaging technique requires the preparation of a positron-emitting radiolabelled probe or radiotracer and for this purpose, fluorine-18 is becoming, more and more often, the radionuclide of choice (adequate physical and nuclear characteristics and potential wide use and -distribution of fluorine-18-labelled radiopharmaceuticals). Considering chemical structures showing a fluoropyridinyl moiety, nucleophilic heteroaromatic substitution at the ortho-position with no carrier-added [18F]fluoride appears today as the most efficient method for the radiosynthesis of radiotracers and radiopharmaceuticals of high specific radioactivity when compared to homoaromatic-, but also aliphatic, nucleophilic radiofluorination. Like for the aliphatic nucleophilic radiofluorinations, only a good leaving group is required (a halogen, or better a nitro- or a trimethylammonium group). There is no need for an additional strong electron withdrawing substituent for activation of the aromatic ring such as in the homoaromatic nucleophilic radiofluorinations, except if one considers meta fluorination. Nucleophilic heteroaromatic substitution and consequent fluorine-18 incorporation are generally performed in DMSO with the no-carrier-added, activated K[18F]F-K222 complex using conventional heating at a moderately high temperature (120-150 degrees C) or microwave irradiation (100 Watt) for a short period of time (1-2 minutes) and often lead to high radiochemical yields. This review summarizes some of the recent applications of these nucleophilic heteroaromatic substitutions in the pyridine series and highlights its potential in the design (not seldom by hydrogen, hydroxyl or halogen replacement by fluorine) and preparation, of often drug-based, fluorine-18-labelled radiotracers and radiopharmaceuticals of high specific radioactivity for PET imaging. PMID- 16250852 TI - PET and SPECT exploration of central monoaminergic transporters for the development of new drugs and treatments in brain disorders. AB - Membrane and vesicular monoaminergic transporters, responsible for the homeostasis of neurotransmitter pools at nerve endings, are very involved in the physiology and diseases of central nervous system. Recent progresses of cerebral molecular imaging using SPECT and PET methods allow the extend of in vivo exploration of these transporters. For this aim, an increasing number of radiopharmaceuticals labelled with [123I], [99mTc], [11C] or [18F] have been developed such as cocaine derivatives for the DAT, compounds from the diphenyl sulfide family for the SERT, and dihydrotetrabenazine derivatives for the VMAT2. These functional imaging methods can be very useful in several neurological and psychiatric disorders which involve the monoaminergic neurotransmission systems such as Parkinson's disease, ADHD, depression and autism. For example, the DAT is a specific index of the density of dopaminergic endings which progressively degenerate in Parkinson's disease. In vivo exploration of this transporter can therefore be a relevant way (i) to realize an early detection of the loss of dopaminergic neurons, (ii) to assess the progression of the disease, (iii) to validate and improve the efficacy of new therapeutic strategies such as neuroprotection and neuroreparation. In all, the extend of in vivo exploration of monoamine transporters will allow great progress for (1) knowledge of physiopathological mechanisms of brain disorders, (2) early diagnosis of cerebral dysfunctions, allowing early use of new therapies, (3) selection of homogenous classes of subjects for therapeutic assays, (4) objectiveness of drug-molecular target interaction, (5) follow-up of disease evolution and treatment. PMID- 16250853 TI - Small animal imaging in drug development. AB - Better mechanistic understanding of disease through mapping of the human and mouse genomes enables rethinking of human infirmity. In the case of cancer, e.g., we may begin to associate disease states with their underlying genetic defects rather than with the organ system involved. That will enable more selective, nontoxic therapies in patients who are genetically predisposed to respond to them. Because one of the major goals of molecular imaging research is to interrogate gene expression noninvasively, it can impact greatly on that process. Most of molecular imaging research is undertaken in small animals, which provide a conduit between in vitro studies and human clinical imaging. We are fortunate to be able to manipulate small animals genetically, and to have increasingly better models of human disease. The ability to study those animals noninvasively and quantitatively with new, high-resolution imaging devices provides the most relevant milieu in which to find and examine new therapies. PMID- 16250854 TI - Perspectives of new antihypertensive drugs. PMID- 16250856 TI - Vasopeptidase inhibition for blood pressure control: emerging experience. AB - Vasopeptidase inhibition is a novel treatment approach in cardiovascular disease such as hypertension and heart failure. Since the inhibition of the angiotensin converting enzyme (ACE) turned out to represent a very successful principle in the treatment of hypertension in numerous large scale clinical studies, their results encouraged attempts to inhibit other key enzymes in the regulation of vascular tone as well--such as the neutral endopeptidase (NEP). Similar to ACE, NEP is an endothelial cell surface metalloproteinase, which is involved in the degradation of several regulatory peptides including the natriuretic peptides and thus augments vasodilatation and natriuresis through increased levels of atrial natriuretic peptide (ANP). By simultaneous inhibition of the RAS and potentiation of the natriuretic peptide system, combined NEP/ACE inhibitors--the so called vasopeptidase inhibitors--reduce vasoconstriction and enhance vasodilatation, therefore, decreasing peripheral vascular resistance and blood pressure. Based on these considerations, numerous preclinicial studies with vasopeptidase inhibitors were performed and revealed promising results in experimental hypertension. Correspondingly, large scale clinical studies in patients with hypertension are on the way. Their preliminary results indicate that combined inhibition of ACE and NEP by vasopeptidase inhibitors represents an effective strategy in the treatment of hypertension and other cardiovascular disease such as heart failure. However, clinical data also suggest that the incidence of angioedema may increase on vasopeptidase inhibition. Therefore, careful evaluation of the safety of this promising therapeutic principle in large scale clinical studies is mandatory before vasopeptidase inhibition may be considered a novel option in the treatment of cardiovascular disease. PMID- 16250855 TI - The renin-angiotensin systems: evolving pharmacological perspectives for cerebroprotection. AB - During the last 20 years, the renin-angiotensin system (RAS) has become an increasingly important focus of basic and clinical cardiovascular research. One main conceptual step forward was made with the discovery of a tissue RAS and the understanding of its critical pathophysiological role in atherogenesis and plaque destabilisation. Major effort to find new strategies for blocking the RAS has produced new classes of drugs which were expected to be clinically important in the management of hypertension and heart failure. As landmark clinical studies have demonstrated that inhibition of the RAS significantly reduces morbidity and mortality from coronary heart disease, myocardial infarction and heart failure, the concept has rapidly emerged that blocking the RAS was the strategy of choice for preventing cardiovascular diseases. More recently, basic research has however continuously extended our understanding of the complexity of the systemic and tissue RASs, that can no longer be viewed as one-way streets in which one single effector, angiotensin II acts solely through its major (AT1) receptor. Meanwhile, clinical trials have challenged the concept that blocking the RAS is the most effective preventive strategy for all patients and all target organs. Consistent with the recent understanding that the RAS encompasses a number of distinct effectors acting through different receptors to promote opposite effects, a growing body of basic and clinical evidence suggests that blunting the RAS is a double-edge sword, with beneficial effects counterbalanced by deleterious ones, resulting in a net effect that critically depends on the experimental conditions, or the clinical characteristics of the study population. Of particular clinical relevance, a number of clinical trials point to the somewhat provocative conclusion that beyond their blood pressure lowering effect antihypertensive drugs that decrease angiotensin II formation are less stroke protective than the ones that increase angiotensin levels. This review focuses on the recent experimental evidence demonstrating that angiotensin II and its derivatives acting through the non-AT1 receptors are involved in protective mechanisms against cerebral ischaemia and discusses in the light of the recent large cardiovascular prevention trials the clinical relevance of this new concept. The perspective of a renewal of therapeutical strategies to optimise the prevention of target organ damage and perhaps even some of the diseases of ageing, such as loss of cognitive function is emphasised. PMID- 16250857 TI - Ouabain antagonists as antihypertensive agents. AB - The evidence that high levels of endogenous ouabain (EO), a closely related isomer of ouabain, are implicated in human hypertension and cardiac hypertrophy and failure stimulated the pharmacological research for developing novel anti hypertensive agents active as ouabain antagonists. The pathogenetic mechanisms through which increased EO levels affect cardiovascular system involve the modulation of Na-K ATPase, the key enzyme responsible for renal tubular sodium reabsorption and the activation of signalling transduction pathways implicated in growth-related gene transcription. By studying both genetic and experimental rat models of hypertension and comparing them with humans, our group has demonstrated that elevated levels of circulating EO and the genetic polymorphism of the cytoskeletal protein adducin associate with hypertension and high renal Na-K pump activity. Ouabain itself induces hypertension and up-regulates renal Na-K pump when chronically infused at low doses into rats (OS). In renal cultured cells, either incubated for several days with nanomolar concentrations of ouabain or transfected with the hypertensive adducin genetic variant, the Na-K pump results enhanced. Moreover, both EO and adducin polymorphism affect cardiac complications associated to hypertension, the former through the activation of a signalling transduction pathway. As a consequence, a compound able to interact with the cellular and molecular alterations, sustained by EO or mutated adducin, may represent the suitable treatment for those patients in whom these mechanisms are at work. A new antihypertensive compound, PST 2238, that selectively antagonises the pressor effect and the alteration of renal Na-K pump, sustained both by ouabain and adducin polymorphism, is described. A selective ability of PST 2238 to antagonise the ouabain-induced organ hypertrophy is also documented. The specificity of PST 2238 mechanism of action is supported by the absence of interactions with receptors or hormones involved in blood pressure regulation and by the lack of diuretic activity and diuretic-associated side effects. It is concluded that this compound could be useful for the treatment of those forms of essential hypertension in which renal Na handling alterations and cardiac complications are associated with either increased EO levels and/or adducin polymorphism. PMID- 16250858 TI - Nitric oxide pathway as new drug targets for refractory hypertension. AB - Nitric oxide (NO) is thought to reduce blood pressure by evoking vasodilation either directly by causing relaxation of vascular smooth muscle or indirectly by acting in the rostral brainstem to reduce central sympathetic outflow, which decreases the release of norepinephrine from sympathetic nerve terminals. An increasingly large body of literature suggests that alterations in the NO system may play an important role in the development or maintenance of clinical hypertension. As proof of concept, pharmacological inhibition of nitric oxide synthase (NOS) in humans and animals causes moderate to severe hypertension. Certain forms of secondary hypertension are accompanied by the accumulation of endogenous NOS inhibitors, which may contribute to the development of hypertension. Furthermore, targeted disruption of the endothelial isoform of NOS in mice causes moderate hypertension, implying that hypertension may also develop from reductions in NOS expression. These gene knockout studies in animals have initiated the search for single nucleotide polymorphisms in human NOS genes, which could potentially lead to decreases in NOS protein expression. Conversely, increases in NOS expression or NO production have been linked with several commonly used cardiovascular therapies, including exercise training and the use of both statins and angiotensin-converting enzyme inhibitors. Finally, increases in the production of oxidants such as superoxide anion can lead to the inactivation of NO, thereby reducing NO bioavailability. Thus, alterations in the expression or activity of NOS or in the availability of NO have the potential to play a causal role in clinical hypertension. The purpose of this article is to show how emerging basic research on the NO pathway is elucidating novel antihypertensive drug targets that are on the cusp of clinical application. PMID- 16250859 TI - Role of calmodulin in platelet receptor function. AB - Platelet glycoprotein (GP)Ib-IX-V and GPVI are unique platelet receptors that bind von Willebrand factor or collagen, respectively, and control the initial interaction of circulating platelets with the blood vessel wall in physiology (hemostasis) or pathology (heart attack or stroke). Engagement of GPIbalpha (the major ligand-binding subunit of GPIb-IX-V) by von Willebrand factor or GPVI by collagen, leads to mobilization of cytosolic Ca2+, secretion of platelet agonists such as ADP, cytoskeletal changes, and activation of the platelet integrin alphaIIbbeta3 that mediates von Willebrand factor- or fibrinogen-dependent platelet aggregation. Recent evidence suggests the cytosolic regulatory protein, calmodulin, plays a central role in regulating GPVI or GPIb-IX-V: first, calmodulin directly binds to conserved, juxtamembrane motifs within cytoplasmic domains of both GPVI and GPIb-IX-V (GPIbbeta and GPV subunits) on resting platelets, interactions that dissociate upon platelet activation; second, an intact calmodulin-binding site within GPVI in transfected cells is required for CaCa2+ signaling, but not for GPVI-dependent pathways involving Src family kinases or co-associated FcRgamma-chain; and third, calmodulin regulates metalloproteinase-dependent ectodomain shedding of GPVI and GPV from human platelets. Other vascular cell adhesion receptors, i.e. leukocyte L-selectin, or PECAM-1 (platelet-endothelial cell adhesion molecule-1), also bind calmodulin within the juxtamembrane region of their cytoplasmic tails, an interaction involved in their proteolytic regulation. Further studies should define the precise functional role of calmodulin in thrombus formation initiated by GPIb-IX V or GPVI. PMID- 16250860 TI - Homocysteine-lowering treatment in coronary heart disease. AB - Homocysteine (Hcy) is a sulphur-containing amino acid product of methionine's metabolism. Hyperhomocysteinemia (HHcy) is considered an independent risk factor for cardiovascular (CV) disease, at least in high-risk patients. In fact, evidence indicates that although mild HHcy may be regarded as a minor risk factor for CHD in low-risk patients, it can play a role in triggering new events in patients with known CHD, also by interacting with the "classical" CV risk factors. This is of much interest because HHcy represents a correctable risk factor, inasmuch vitamin supplementation as has been shown to effectively lower total homocysteine plasma levels (tHcy). While case-control and cross-sectional studies have consistently demonstrated an association of HHcy with CV disease, prospective studies have given conflicting results. Moreover, the effect of the homocysteine-lowering treatment in preventing CV events is still under debate. Thus, it remains unclear which patients should be screened for HHcy and which ones should be treated to lower tHcy. In this paper we shall report and discuss knowledge on the potential role of HHcy in the development of CHD and on the benefits due to tHcy-lowering treatment with vitamins. PMID- 16250861 TI - Anemia in chronic heart failure--clinical and prognostic significance. AB - Anemia is a common finding in chronic heart failure (CHF). Anemia develops due to CHF, but is also known to cause heart failure. Patients with CHF are limited by exercise capacity and fatigue. Low hemoglobin concentration can account for both and may substantially contribute to the symptoms of CHF. Increasing severity of CHF is associated with a higher frequency of anemia, which also becomes clinically more relevant. Anemia has been shown to predict impaired survival in CHF, independent of established prognostic markers. There are many potential reasons for development of anemia in CHF, such as bone marrow depression, reduced intestinal iron uptake and hemodilution as a consequence of sodium and water retention. In most cases, however, anemia in CHF should be viewed as "anemia of chronic illness", being the result of a combination of many factors related to the disease, particularly chronic inflammation. The option of therapeutically targeting anemia in CHF is an intriguing novel approach to improve morbidity and potentially mortality in these patients. PMID- 16250862 TI - Recent progress in signal transduction research of the angiotensin II type-1 receptor: protein kinases, vascular dysfunction and structural requirement. AB - Accumulating evidence strongly implicates the critical roles of intracellular signaling of angiotensin II (AngII) in mediating cardiovascular diseases such as hypertension, atherosclerosis, and restenosis after vascular injury. The importance of AngII signals has also been reported in endothelial dysfunction and insulin resistance, two strong predictors of cardiovascular disease. Through its G protein-coupled AngII type-1 receptor (AT1), AngII activates various intracellular protein kinases, such as receptor or non-receptor tyrosine kinases and serine/threonine kinases. Activation of these kinases requires both G protein dependent and independent pathways, reactive oxygen species and a metalloprotease, and each kinase could be involved specifically in mediating pathophysiological function of the AT1 receptor target organs. In fact, some of the kinases are indispensable for AngII-induced hypertrophy and migration. The role of these AT1-activated kinases in mediating vascular remodeling, vascular contractility, endothelial dysfunction, and insulin resistance will be discussed in this review. In addition, the AT1 receptor undergoes rapid phosphorylation, desensitization, and internalization upon AngII stimulation. Recent studies with site-directed mutagenesis of the AT1 receptor not only elucidated a G protein interaction and desensitization of the receptor, but also demonstrated a structural requirement of the receptor for downstream signal transduction. Thus, AT1 mutants have provided an excellent means to examine the mechanism of signal transduction and their significance in mediating AngII function. Taken together, in this review, we will focus our discussion on the recent findings of the signal transduction research elucidating novel signaling mechanisms of the AT1 receptor that are relevant to the vascular pathophysiology of AngII. PMID- 16250863 TI - Strategic leukofiltration in cardiac surgery. AB - Systemic inflammatory response syndrome (SIRS) with activation of molecular cascades, cell activation, accumulation of interstitial fluid, organ dysfunction and, occasionally, organ failure is still a commonly recognized consequence of cardiac surgery. SIRS leads to costly complications and several strategies intended to ameliorate the symptoms that have been studied, including leukocyte reduction, using filtration. Although, the body of work suggests that leukoreduction attenuates SIRS, discrepancies remain within the literature. The recent literature is reviewed highlighting the areas where concordance is lacking. In our study, on the basis of indirect indicators of SIRS, platelet function by thromboelastography biomaterial evaluation by light and scanning electron microscopy, we present our conclusions regarding clinical outcomes and the role of leukofiltration. PMID- 16250864 TI - Modulators of ion channels activated by hypotonic swelling in cardiomyocytes: new perspectives for pharmacological treatment of life-threatening arrhythmias. AB - This review highlights the most important research data related to membrane potential and current changes in cardiomyocytes during hypotonic stress. Relative decrease in osmolarity in extracellular compartment (due to accumulation of metabolic products in the cells) during acute episodes of ischemia in the heart muscle leads to cell swelling caused by water entering the cells. Such condition starts regulatory volume decrease (RDV) in cardiomyocytes--a process involving activation of various ion channels. It seems to be the crucial proarrhythmic mechanism in ischemic heart, probably very often responsible for sudden cardiac death. Understanding of electrophysiological changes during hypotonic stress of cardiomyocytes is a basis for appropriate pharmacological intervention preventing serious arrhythmias. For instance, outwardly rectifying swelling-induced chloride currents (IClswell), inwardly rectifying non-selective cation current (INSC) and slow component of delayed rectifier K+ currents (IKs) are activated during hypotonic stress in ventricular myocytes and substances like anthracene-9 carboxylic acid (9-AC), chromanol (293B) and gadolinium (Gd3+), able to modulate former channels, should be considered to be potential antiarrhythmic drugs in the near future. PMID- 16250865 TI - Fibrinolysis at the fluid-solid interface of thrombi. AB - Thrombolysis is conventionally regarded as dissolution of the fibrin matrix of thrombi by plasmin, a protease generated by plasminogen activators from its inactive precursor, plasminogen. Typically plasminogen activation occurs on the surface of the clot, where fibrin behaves as a cofactor in this process, and plasmin also initiates its proteolytic action at the fluid-solid interface. Although the basic reactions of the plasminogen/plasmin system in fluid phase are well characterized in terms of classical enzymology, they cannot explain completely the interfacial fibrinolytic events. Recently new methods have been introduced for quantitative evaluation of plasminogen activation on gel-phase fibrin and heterogenous-phase proteolysis, an overview of the new methodology is presented. Following formation of an interfacial lytic zone, fibrin dissolution proceeds through propagation of this zone to the core of the clot, which depends on diffusion and permeation phenomena affected by the composition of thrombi. Phospholipids (originating from platelets) form a diffusion barrier to the thrombolytic agents and also bind some of them; structural cellular proteins (namely myosin) interact with the fibrin fibers masking their cofactor and plasmin-cleavage sites. The contribution of these recent findings to our understanding of the limitations of current thrombolytic therapy is discussed. Finally, attention is focused on the termination of thrombus-associated proteolytic action in an environment abundant in proteinase inhibitors. Thus, combining together the interfacial events in the initiation, progress and termination of thrombolysis, a concept for modeling the thrombus as a temporary fibrinolytic compartment is presented. PMID- 16250866 TI - Pharmacogenetics in cardiovascular antithrombotic therapy. AB - Thrombosis is the most important underlying mechanism of coronary heart disease and embolic stroke. Therefore, antithrombotic therapy is commonly used in cardiovascular diseases. Unfortunately, the benefits are limited, and an important proportion of treated patients will suffer a new thrombotic event. Lack of clinical benefits may be related to heterogeneous response to antithrombotic treatment among individuals (inter-individual heterogeneity). Few factors have been identified to be involved in this inter-individual heterogeneity. Recently, pharmacogenetic has emerged as a new field in medicine that tries to identify gene variants able to explain the heterogeneity in patient's response to a drug. Polymorphisms affecting disposition, metabolism, transporters or targets of the drug could modify the individual response to one therapy, and probably its side effects. The present review article explores the genetic influence on antithrombotic drug efficacy, analysing the modulating role of different polymorphisms on individuals' response to drugs commonly used in current day practice. PMID- 16250867 TI - Thrombocytopenia in HIV disease: clinical relevance, physiopathology and management. AB - HIV infection is associated with a myriad of hematopoietic abnormalities. Thrombocytopenia (TCP), the condition in which platelet counts fall below 150x10(3)/mm3 in two or more consecutive platelet counts, is a condition frequently seen in HIV infected individuals regardless of HIV status, gender, or age. Having recently been associated with rapid disease progression, and by complicating the management of AIDS patients, thrombocytopenia has become a medical challenge, highlighting the urgent need for evidence-based treatment protocols in this area. Due to the physiopathology of HIV, therapeutic options currently available for TCP in this already vulnerable population are severely limited. Whereas clinicians often intervene to prevent life-threatening, thrombocytopenia-associated outcomes in the general population, there is no intervention protocol: for the HIV subjects. Management of the condition seems to be the norm for these individuals. As a result, thrombocytopenia in HIV is a subject that is in urgent need of re-examination. In this review, the importance of thrombocytopenia and current knowledge regarding the physiopathology of HIV associated thrombocytopenia is discussed, and an overview of current and under investigation treatment approaches to this adverse hematological condition is provided. PMID- 16250868 TI - Clinical determination of the severity of metabolic syndrome: preheparin lipoprotein lipase mass as a new marker of metabolic syndrome. AB - The severity of metabolic syndrome depends on the degree of insulin resistance. However, currently there is no adequate clinical marker for quantitative analysis of insulin resistance. A small quantity of lipoprotein lipase (LPL) protein, which is an inactive form and commonly called 'preheparin LPL mass', exists in serum and is detected by a sensitive immunoassay system. Recent studies have reported the clinical significance of serum preheparin LPL mass levels in various aspects. For example, preheparin LPL mass is negatively related to serum triglyceride and positively related to HDL-cholesterol, is low in type 2 diabetes mellitus, is increased by administration of insulin sensitizer, and shows an inverse relationship with visceral adiposity. Furthermore, preheparin LPL mass level is significantly lower in patients with coronary atherosclerosis compared to patients with no lesion, and correlates negatively with the severity of these lesions. From these reports, preheparin LPL mass may be considered to be the most important quantitative indicator of insulin resistance of the whole body. PMID- 16250869 TI - Angiotensin-(1-7): blood, heart, and blood vessels. AB - In the past few years, there has been a growing interest in the heptapeptide Angiotensin(Ang)-(1-7), mainly because of its ability to counter regulate many of Ang II actions. Furthermore, heart and blood vessels are important target tissues for Ang-(1-7) formation and actions. The introduction of novel tools, such as the Ang-(1-7) antagonists, A-779 and D-pro7-Ang-(1-7), the Ang-(1-7) agonist AVE 0991, transgenic rats TGR(A-1-7)3292, and use of liposome-encapsulated Ang-(1-7) for evaluating the biochemical and functional role of Ang-(1-7), have produced a great impact in this field of research. Moreover, the recent identification of the Ang-(1-7)-forming enzyme ACE2 and of the Ang-(1-7) receptor Mas will allow important advances in our understanding of the physiological and pathological role of this peptide. In this review, we will discuss the current knowledge concerning the biological effects of Ang-(1-7) in the blood, heart, and blood vessels. In addition, we will highlight the possible applications of agonists of its receptor as therapeutic agents in cardiovascular and related diseases. PMID- 16250871 TI - Naturally-occurring xanthones: recent developments. AB - A literature survey covering the report of naturally occurring xanthones from January 2000 to December 2004, with 219 references, is presented in this review. Among 515 xanthones reported in this period, 278 were new natural xanthones. These xanthones have been identified from 20 families of higher plants (122 species in 44 genera), fungi (19 species) and lichens (3 species). The structural formulas of 368 identified xanthones, their distribution and a brief mention of their biological properties are also included. PMID- 16250872 TI - Synthesis of xanthones: an overview. AB - Among the known synthetic routes to obtain xanthones, the Grover, Shah, and Shah reaction, the cyclodehydration of 2, 2'-dihydroxybenzophenones and electrophilic cycloacylation of 2-aryloxybenzoic acids are the most popular methods. Due to important biological applications of xanthones, some synthetic strategies leading to more complex derivatives have been widely explored in the past years. Thus, the purpose of this review is to report some recent improvements of the classical synthetic methods as well as of some non-classical methods to obtain simple oxygenated xanthones. The strategies for introduction of substituents into the xanthonic nucleus are also summarized. Furthermore, different approaches used to synthesize complex structures, with an emphasis on the total synthesis of bioactive natural products, accomplished in the last twenty years, are also discussed. Besides the synthesis of xanthones, the reactivity of the xanthonic nucleus and its role as a key intermediate for the synthesis of other important classes of compounds are also highlighted. PMID- 16250873 TI - Structure elucidation of xanthone derivatives: studies of nuclear magnetic resonance spectroscopy. AB - 1H and 13C NMR spectra remain the first tool used by chemists to perform the structure elucidation of their products on a routine basis. It is common to provide NMR data on both proton and carbon spectra based on one-dimensional experiments, but often only proton resonances are assigned. The increasing complexity of natural compounds and their synthetic related derivatives imply the use of some more recent 1D and 2D NMR techniques. The purpose of this review is to describe the main NMR features of the most common and important classes of xanthones and also to discuss the application of several 1D and 2D NMR techniques in the structure elucidation of these compounds. A brief discussion of these NMR techniques from the point of view of structure elucidation of organic compounds will also be considered. The calculated NMR chemical shifts in the structure elucidation of xanthones and the use of NMR to study their mode of action in biological activities will be also described. PMID- 16250874 TI - Xanthones--a structural perspective. AB - Xanthones, synthesized or isolated from a natural source, display a wide range of biological and pharmacological activities. In a few cases, their chemical characterization has involved the structure elucidation by single crystal X-ray diffraction. The purpose of this review is to assess in detail this three dimensional structural data, and thus contribute to a better understanding of the molecular mechanisms involved in the different biological activities presented by xanthones. PMID- 16250875 TI - Xanthone derivatives: new insights in biological activities. AB - Xanthones or xanthen-9H-ones (dibenzo-gamma-pirone) comprise an important class of oxygenated heterocycles whose role is well-known in Medicinal Chemistry. The biological activities of this class of compounds are associated with their tricyclic scaffold but vary depending on the nature and/or position of the different substituents. In this review, an array of biological/pharmacological effects is presented for both natural and synthetic xanthone derivatives, with an emphasis on some significant studies on structure-activity relationships. The antitumor activity of some xanthones as well as the related targets, particularly PKC modulation studies, is also discussed in detail. Examples of the "hit" compounds involved in cancer therapy, namely DMXAA, psorospermin, mangiferin, norathyriol, mangostins, and AH6809, a prostanoid receptor antagonist, are also mentioned. Finally, a historical perspective of these xanthonic derivatives, their relevance as therapeutic agents and/or their uses as pharmacological tools and as extract components in folk medicine are also highlighted. PMID- 16250876 TI - Xanthones as antimalarial agents: discovery, mode of action, and optimization. AB - It is believed that at no time in the history of the human race malaria has been absent. This disease, which is caused by protozoa of the genus Plasmodium, in all likelihood has been responsible for the death of about half of all people who ever lived. Even today, after attempts at intervention on a worldwide scale, malaria remains the most significant parasitic disease in the tropics and sub tropics, where it causes at least 500 million clinical episodes and claims 1.5 million lives each year, mostly young children and pregnant women. Widespread resistance to the best and least expensive antimalarials, chloroquine and S/P (i.e., a combination of sulfadoxine and pyrimethamine), combined with an increasing tolerance to insecticides in the mosquito vector, threaten a global malaria tragedy unless new countermeasures are developed. For malaria therapy, the great panacea would be the development of a long-lasting vaccine, but until this becomes a reality, people living in and traveling to endemic regions must rely on a dwindling cache of more expensive drugs; many beyond the economic reach of impoverished people living in malarious regions of the world. Our course to recognition of xanthones as potential antimalarial agents took a rather circuitous route, involving both serendipity and empiricism, and is described together with mechanistic details of drug action. From a chance encounter with a sea urchin collected near the city of Cannon Beach on the Oregon coast to naturally occurring and functionalized xanthones, it is revealed how these compounds target the Plasmodium parasite's most vulnerable feature--the digestive vacuole. PMID- 16250877 TI - Development of anti-HIV agents targeting dynamic supramolecular mechanism: entry and fusion inhibitors based on CXCR4/CCR5 antagonists and gp41-C34-remodeling peptides. AB - A molecular mechanism involved both in HIV-entry and -fusion steps has been disclosed in detail: The interaction of an HIV envelope protein, gp120, with chemokine receptors, CXCR4 and CCR5, which were identified as major co-receptors in association with CD4, triggers conformational changes in the gp120-gp41 (another envelope protein) complex, and subsequently forms the trimer-of-hairpins structure of gp41 followed by virus-cell membrane fusion. The elucidation of the above dynamic supramolecular mechanism in HIV-entry and -fusion has provided insights into new type of drugs that can block HIV infection. Based on this, we have developed not only coreceptor antagonists (1) but also fusion inhibitors (2). (1) Potent CXCR4 antagonists, T22 and T140, have been developed through the structure-activity relationship studies on tachyplesins and polyphemusins that are horseshoe crabs' antimicrobial peptides. T22, which was initially found to bind gp120 and CD4, and T140 selectively suppress T-cell line-tropic HIV-1 (X4 HIV-1) entry based on their specific binding to CXCR4. Furthermore, molecular size reduction of T140 using cyclic pentapeptide templates brought us to find low molecular weight CXCR4 antagonists, such as FC131. (2) Artificial remodeling of a gp41 fragment, C34, has led to development of strong inhibitors of HIV-fusion into cells. These fusion inhibitors effectively block the formation of the trimer of-hairpins structure of gp41. HIV-entry/fusion inhibitors such as CXCR4 antagonists and C34 analogs would improve the clinical chemotherapy of AIDS and HIV-infected patients. This review article focuses on our recent research on the development of the above two types of inhibitors, including comparative studies with several CXCR4 antagonists besides T22/T140-related compounds and other fusion inhibitors such as Fuzeon (T-20). PMID- 16250879 TI - AIDS related viruses, their association with leukemia, and Raf signaling. AB - Leukemia is characterized by the production of an excessive number of abnormal white blood cells. Over time, this expanding population of poorly/non- functional white blood cells overwhelms the normal function of the body's blood and immune systems. DNA translocations have been found common to leukemia, including Raf mutations. While the cause of leukemia is not known, several risk factors have been identified. In this review, we present an update on the role of AIDS related viruses as an etiology for leukemia. Human immunodeficiency virus-1 and -2 (HIV 1; -2) are the cause for the development of acquired immune deficiency syndrome (AIDS). Epstein-Barr virus (EBV), human cytomegalovirus (HCMV), Human papillomavirus (HPV), and Kaposi's sarcoma-associated herpesvirus (KSHV) are specifically implicated in AIDS associated malignancies. However, there are other viruses that are associated to a lesser extent with the AIDS condition and they are Human T-cell leukemia virus-1 (HTLV-1), hepatitis B virus (HBV), hepatitis C virus (HCV), and human herpesvirus-6 (HHV-6). Of these viruses, HTLV-1 has been etiologically associated with leukemia. Recent evidence suggests that EBV, HBV, HCV, and KSHV may also play a role in the development of some types of leukemia. Raf signaling has been shown to aid in the infection and pathogenesis of many of these viruses, making Raf pathway components good potential targets for the treatment of leukemia induced by AIDS related viruses. PMID- 16250878 TI - Transendothelial migration of monocytes: the underlying molecular mechanisms and consequences of HIV-1 infection. AB - Migration of monocytes from the bloodstream across vascular endothelium is required for routine immunological surveillance of tissues and their entry into inflamed sites. Transendothelial migration of monocytes initially involves tethering of cells to the endothelium, followed by loose rolling along the vascular surface, firm adhesion to the endothelium and diapedesis between the tightly apposing endothelial cells. A number of adhesion molecules are involved in this process. Monocyte rolling can be mediated by selectins and their ligands, or alpha(4)beta(1) integrin interacting with endothelial VCAM-1. On the apical surface of the endothelial cell, bound chemokines (eg. MCP-1, MIP-1alpha/beta) can activate leukocyte beta(2) integrins for tight adhesion to ICAM-1 and -2. Diapedesis by monocytes occurs through interaction between PECAM-1 on both the monocyte and the endothelial cells, followed by similar homophilic adhesion via CD99. After penetration of the endothelial basement membrane, monocytes migrate through the extracellular matrix of the tissues where they may differentiate into tissue macrophages and/or migrate to sites of inflammation. Additionally, monocytes in the tissues may traffic to the lymphatics or back into the bloodstream, both of which involve basal to apical (reverse) transendothelial migration, possibly mediated by tissue factor and p-glycoprotein. Monocyte trafficking is of current interest in studies of the pathogenesis of HIV infection, including establishment of viral reservoirs in tissues and sanctuary sites and the development of HIV-related dementia. This review provides insights into the most recent studies on the process of monocyte migration across the vascular endothelium, and changes in migration that can occur during HIV infection. PMID- 16250880 TI - Activation of the RNA-dependent protein kinase (PKR) of lymphocytes by regulatory RNAs: implications for immunomodulation in HIV infection. AB - It has been known for decades that exogenous RNAs are able to induce cytotoxic T lymphocytes (CTLs) and immunological reactivity to a wide variety of antigens. The molecular events responsible for these effects remain unclear for more than two decades. It has been decided to revisit this phenomenon in the light of new concepts that are just emerging in Molecular Biology, such as the regulation of gene expression by noncoding RNAs, named regulatory RNAs. The immunological effects observed in lymphocytes treated with RNAs obtained from lymph nodes of immunized animals with different types of antigens including synthetic peptides of the human immunodeficiency virus type 1 (HIV-1) have been investigated. Our recent results showed that regulatory RNAs are involved in this phenomenon, which is due to the activation of the RNA-dependent protein kinase (PKR) by regulatory RNAs with subsequent activation of the transcription factor NF-kappaB. The RNA dependent protein kinase (PKR) is a serine/threonine kinase and contains two RNA binding domains (RBD-I and RBD-II) within the N-terminal region. PKR is activated by viral double-stranded RNA (dsRNA) and highly structured single-stranded RNAs. This review will focus on the structure and functions of PKR including its role in HIV-1 infection. Special emphasis will be placed on a regulatory RNA, named p9 RNA, isolated from lymphocytes of animals immunized with the synthetic peptide p9 (pol: 476-484) of HIV-1. It was found that the regulatory p9-RNA induces CTLs and production of IFN-gamma. These findings showed for the first time that transcriptional control of gene expression by a regulatory RNA can be mediated by PKR through the activation of the transcription factor NF-kappaB. A model for the mechanism of action of the regulatory p9-RNA responsible for the production of IFN-gamma is proposed. Elucidating the molecular mechanism of p9-RNA may contribute to determining the rationale for the use of this regulatory RNA as an immunomodulator in HIV-infected patients. PMID- 16250881 TI - Standing in the way of eradication: HIV-1 infection and treatment in the male genital tract. AB - As a result of the introduction of the Highly Active Antiretroviral Therapy (HAART) many HIV-1 infected individuals are able to live an improved and extended life style that can include the prospect of having children. Problematically, the male reproductive organs may contribute infected cells and free viral particles to semen in these individuals, increasing the risk of infection from the HIV-1 positive male to the mother and ultimately to the offspring. Though autopsies of AIDS infected individuals have taught us a great deal about specific cell loss and the deterioration of male organs in this setting, it is not clear whether the damage is due to specific targeting of these cells and tissues by HIV-1 or an indirect consequence of viral dissemination in the later stages of infection. Due to lack of access to these organs in the early stages of the disease, little is known about the progression and pathogenesis of the infection within them, particularly during the asymptomatic stage. The molecular and cellular aspects of transmission of this virus remain unclear. Although assisted reproductive techniques have been successfully used to achieve pregnancies in discordant couples in the developed world, investigating the mechanism of the spread of HIV 1 in the cells and tissues of the male reproductive tract remains a critical task, not only to improve our understanding, but also to enable the design of suitable treatment for the eradication of the infection from this potential sanctuary site. In this review, we discuss possible mechanisms by which infection of the male genital tract (MGT) may occur in the context of the anatomy and immunology of the tissues of the male reproductive organs. We revisit the methodology used to evaluate the spread and transmission of HIV-1 from these tissues and pinpoint future directions for study that may provide a better understanding of the underlying mechanisms of HIV-1 transmission by this route. PMID- 16250882 TI - CC and CXC chemokines in breastmilk are associated with mother-to-child HIV-1 transmission. AB - INTRODUCTION: CC and CXC chemokines may play a role in mother-to-child HIV-1 transmission by blocking HIV-1 binding to chemokine receptors and impeding viral entry into cells. METHODS: To define correlates of breastmilk chemokines and associations with infant HIV-1 acquisition, chemokines in breastmilk and infant HIV-1 infection risk were assessed in an observational, longitudinal cohort study. We measured MIP-1alpha, MIP-1beta, RANTES, and SDF-1 in month 1 breastmilk specimens from HIV-1-infected women in Nairobi and HIV-1 viral load was calculated in maternal plasma and breastmilk at delivery and 1 month postpartum. Infant infection status was determined at birth and months 1, 3, 6, 9, and 12. RESULTS: Among 281 breastfeeding women, 60 (21%) of their infants acquired HIV-1 during follow-up, 39 (65%) of whom became infected intrapartum or after birth. MIP-1alpha, MIP-1beta, RANTES, and SDF-1 were all positively correlated with breastmilk HIV-1 RNA (P<0.0005). Women with clinical mastitis had 50% higher MIP 1alpha and MIP-1beta levels (P<0.001 and P=0.006, respectively) and women with subclinical mastitis (breastmilk Na(+)/K(+)>1) had approximately 70% higher MIP 1alpha, MIP-1beta and RANTES (P<0.002 for all) compared to women without mastitis. Independent of breastmilk HIV-1, increased MIP-1beta and SDF-1 were associated with reduced risk of infant HIV-1 (RR=0.4; 95% CI 0.2-0.9; P=0.03 and RR=0.5; 95% CI=0.3-0.9; P=0.02, respectively) and increased RANTES was associated with higher transmission risk (RR=2.3; 95% CI 1.1- 5.3; P=0.04). CONCLUSIONS: These observations suggest a complex interplay between virus levels, breastmilk chemokines, and mother-to-child HIV-1 transmission and may provide insight into developing novel strategies to reduce infection across mucosal surfaces. PMID- 16250883 TI - Alternative approach to blood screening using the ExaVir reverse transcriptase activity assay. AB - 408 non-selected samples were obtained from healthy, adult individuals donating blood at the Ethiopian Red Cross Society-National Blood Transfusion Service. All samples were screened for HIV using the Vironostika Ag/Ab test, the Amplicor DNA PCR and examined for the presence of HIV reverse transcriptase (RT) using the ExaVir Load test (version 2). A panel of supplementary tests was used to evaluate the HIV status of the discordant samples and to confirm positivity. One aim was to assess an RT based test for screening for HIV in comparison with other more conventional tests. An HIV-prevalence of 3.4 % (14/408) was found. The Vironostika Ag/Ab test produced 391 negative, and according to the supplementary testing, 14 true- and three false- positive test results. The corresponding figures for the Amplicor DNA PCR test was 384 negative, 14 true- and two extra probably false -positive samples. In addition, the DNA PCR generated eight indeterminate results. The colorimetric version of the ExaVir load test exhibited 100 % specificity and detected RT in 13 of the true positive samples, but failed to detect one sample containing 200 HIV RNA copies /mL. This sample was detectable in the fluorimetric version of the test. The detection of RT activity in addition to the currently used markers would seem to have a potential for use in blood screening. PMID- 16250885 TI - HIV-1 Vif: HIV's weapon against the cellular defense factor APOBEC3G. AB - The human immunodeficiency virus type 1, HIV-1, has long been known to possess the viral infectivity factor, Vif, which supports productive viral replication in non-permissive cells, such as peripheral blood lymphocytes (PBL). In the last few years, Vif function has been elucidated by the finding that it inactivates a cellular anti-viral factor named APOBEC3G. Tremendous progress has been made since the initial observation, reflected in a large number of publications. APOBEC3G represents a novel innate defense mechanism against retroviral infection. It is expressed in non-permissive cells and possesses cytidine deaminase activity. APOBEC3G is encapsidated into viral particles and is transported into the infected cell, where it facilitates the deamination of the cytosine residues in the first strand cDNA intermediate during early steps of HIV infection. Vif counteracts APOBEC3G by direct binding, which mediates its degradation by the ubiquitin-dependent proteasomal pathway. In this review, we will summarize the current knowledge about the structure and function of both proteins, their interaction with each other and the mechanism of Vif-mediated APOBEC3G inactivation. In addition, we will discuss possible interference strategies as potential new drugs against HIV infection. PMID- 16250886 TI - HSV-1-derived recombinant and amplicon vectors for gene transfer and gene therapy. AB - Herpes simplex virus type 1 (HSV-1) is a major human pathogen whose lifestyle is based on a long-term dual interaction with the infected host characterized by the existence of lytic and latent infections. Although in most cell types infection with HSV-1 will induce toxic effects ending in the death of the infected cells, the very deep knowledge we possess on the genetics and molecular biology of HSV-1 has permitted the deletion of most toxic genes and the development of non pathogenic HSV-1-based vectors for gene transfer. Several unique features of HSV 1 make vectors derived from this virus very appealing for preventive or therapeutic gene transfer. These include (i) the very high transgenic capacity of the virus particle, authorizing to convey very large pieces of foreign DNA to the nucleus of mammalian cells, (ii) the genetic complexity of the virus genome, allowing to generate many different types of attenuated vectors possessing oncolytic activity, and (iii) the ability of HSV-1 vectors to invade and establish lifelong non-toxic latent infections in neurons from sensory ganglia and probably in other neurons as well, from where transgenes can be strongly and long-term expressed. Three different classes of vectors can be derived from HSV 1: replication-competent attenuated vectors, replication-incompetent recombinant vectors, and defective helper-dependent vectors known as amplicons. Each of these different vectors attempts to exploit one or more of the above-mentioned features of HSV-1. In this review we will update the current know-how concerning design, construction, and recent applications, as well as the potential and current limitations of the three different classes of HSV-1-based vectors. PMID- 16250884 TI - Rapid size dependent deletion of foreign gene sequences inserted into attenuated HIV-1 upon infection in vivo: implications for vaccine development. AB - Live attenuated HIV vaccines offer a means to introduce exogenous sequences into the viral genome to target the virus elimination in vivo. Foreign genes inserted into the nef region of HIV-1 NL4-3 were found to be rapidly deleted following virus infection and/or replication, in a size dependent manner, in the human fetal Thymus/Liver implants of severe combined immunodeficient mouse (SCID-hu) model. When the murine heat stable antigen (HSA) of 283 bp was substituted into HIV-1 nef region, the viral loads in vivo were comparable to the negative control nef attenuated HIV-1, and the reporter HSA gene was not deleted upon infection. However, the murine Thy1.2 gene (505 bp) substituted into the nef attenuated HIV 1, upon infection and replication, deleted 441 bp in vitro and 437 bp in vivo, of the inserted Thy1.2 gene. When the enhanced green fluorescence protein (eGFP) gene (720 bp) was substituted for nef, virus replication was aborted in vivo in the Thy/Liv implants, as seen by the background levels of viral loads, comparable to mock infected implants, and the eGFP gene was deleted. When the herpes simplex virus thymidine kinase gene, HSV-TK (1.15 kbp), or HSA gene, was substituted into the viral vpr gene, TK but not HSA gene was deleted, upon infection in vitro. Moreover, NL-TKI reporter virus with both intact nef and vpr genes shows deletion of TK gene both in vitro and in vivo. Excision of foreign genes occurred within the exogenous segments but not in the viral own regions. These results suggest that larger "suicide" genes introduced via HIV-1 can be deleted upon infection. However, smaller size nucleotide sequences or genes (approximately 300 bp) inserted in place of viral nef or vpr gene may be used to target the virus or its components, for attack and elimination in vivo, and thus have implications for the development of live attenuated HIV vaccines. PMID- 16250887 TI - The use of gene therapy tools in reproductive immunology research. AB - Mammalian pregnancy is a complex phenomenon allowing the maternal immune system to support its allogeneic fetus, while still being effective against pathogens. Gene therapy approaches have the potential to treat devastating inherited diseases for which there is a little hope of finding a conventional cure. In reproductive medicine, experimental trials have been made so far only for correcting gene defects in utero. The use of gene therapy for improving pregnancy rate success or avoiding pregnancy-related diseases i.e. miscarriage or pre eclampsia, remains a very distant goal with unresolved moral and ethical aspects. However, gene therapy may help determining the role of several genes in supporting fetal growth and/or avoiding its rejection experimentally and might further help to identify new targets of intervention. Gene therapy strategies to avoid fetal rejection may include the transfer and expression of cyto-protective molecules locally at the fetal-placental interface. In addition, the ex-vivo genetic modification of immune cells for tolerance induction is a novel and tempting approach. In this regard, we have confirmed the role of the cyto protective and immunomodulatory molecule Heme Oxygenase-1 (HO-1), by treating animals undergoing abortion with an adenovirus coding for HO-1. Since the sole application of a control vector did not provoke deleterious effects in pregnancy outcome, we propose the use of experimental gene therapy for unveiling molecular and cellular pathways leading to pregnancy success. PMID- 16250888 TI - RNA splicing manipulation: strategies to modify gene expression for a variety of therapeutic outcomes. AB - Antisense oligonucleotides initially offered great hope as specific compounds to modify gene expression, primarily through RNaseH induced degradation of the target transcript. Expansion of the field led to new chemistries capable of invoking different mechanisms, including suppression of protein synthesis by translational blockade, and there is now a major interest in downregulation of gene expression using short interfering RNAs to induce RNA silencing. Naturally occurring microRNAs have been implicated in the regulation of gene expression. This review considers examples of antisense oligonucleotides redirecting the process of exon recognition and intron removal during gene transcript splicing. While suppression of gene expression is necessary to address some conditions, it appears likely that there may be many more clinical applications for antisense oligonucleotides in re-directing splicing patterns. Pre-mRNA splicing is a tightly co-ordinated, multifactorial process, which can be disrupted by antisense oligonucleotides in a highly specific manner, allowing either suppression of aberrant splicing, by-pass of nonsense or frame-shifting mutations or alteration of spliceoform ratios. Manipulation of splicing patterns has been applied to a diverse range of conditions, including beta-thalassemia, Duchenne muscular dystrophy, spinal muscular atrophy and certain cancers. Alternative exon usage has been identified as a major mechanism for generating diversity from a limited repertoire of genes in higher eukaryotes. Considering that up to 75% of all human primary gene transcripts are reported to be alternatively spliced, intervention at the level of pre-mRNA processing is likely to become increasingly significant in the fight against genetic and acquired disorders. PMID- 16250889 TI - Recent advances in the development of adenovirus- and poxvirus-vectored tuberculosis vaccines. AB - Tuberculosis vaccine research began with the search for a vaccine that might be better than, and thus could replace, the current Bacillus Calmette Guerin (BCG) vaccine. Over the last fifteen years or so, intense research effort has led to the identification of a number of novel tuberculosis (TB) vaccines which can be divided into 4 categories: genetically modified mycobacteria, protein, plasmid DNA and viral. However, it is increasingly believed that the current BCG vaccine will continue to be used as a childhood vaccine and that more effort should be directed to developing appropriate boosting vaccines. Mounting evidence suggests that recombinant genetic vaccines, particularly recombinant viral vaccines, are effective in boosting immune activation and protection by BCG vaccination. Since modified vaccinia virus Ankara (MVA)- and adenovirus-vectored TB vaccines have been most extensively studied, this review will focus on recent advances in the development and applications of these two viral TB vaccines. PMID- 16250891 TI - Genetic idiotypic and tumor cell-based vaccine strategies for indolent non Hodgkin's lymphoma. AB - B cell malignancies express a clear tumor-specific antigen (B cell immunoglobulin variable regions) known as idiotype (Id). It is now possible to immunize patients against autologous Id generating humoral and cellular immune responses that correlate with clinical and molecular remissions and the possibility of improved disease-free survival. In its present form, however, individual vaccine preparation by generating heterohybridomas is a technical and financial challenge. DNA vaccination provides a unique opportunity to streamline individual vaccine manufacture by circumventing the need for protein purification. DNA fusion vaccines have been developed in which genetic carriers promote adaptive immunity against the attached Id. Such carriers can specifically bind receptors on dendritic cells (DC) for targeted antigen delivery, or supply high levels of T cell help. Ideally, the carrier should be able to activate innate immunity to enhance the antigen-presenting capacity of DC. The correlates of immunity may vary depending upon the genetic carrier used. Translation to patients has begun with preliminary evidence of Id-specific immune responses. An alternative vaccination strategy that allows for the potential to vaccinate against multiple tumor antigens without the need to identify individual antigens is based on tumor cells themselves to be used as vaccine. To this purpose, however, each patient's tumor cells must be genetically modified to increase their immunogenicity. To overcome the technical limitations inherent with a fully autologous approach, strategies have been devised where a universal, genetically modified bystander cells is expected to provide the immunoenhancing cytokines to allow immune recognition of unmodified patients' tumor cells. PMID- 16250890 TI - Animal models for growth hormone gene therapy. AB - Treatment of growth hormone (GH) deficiency via parenteral administration of recombinant hGH has greatly benefited from recombinant DNA technology allowing production of practically unlimited amounts of the pure hormone. However, an alternative approach that may lead to correction of the clinical defect is presented by hGH gene transfer into somatic cells of the patient, either ex vivo or in vivo. This approach has not only the potential advantage of circumventing repetitive injections of the hormone and its laborious isolation and purification processes, but can also, in principle, provide a mechanism of hormone delivery that resembles the natural process. GH gene therapy has not reached the clinics yet, but several interesting and promising animal models for this treatment have been developed and studied. They are not only potentially useful for elucidation of the still unresolved mechanism of sustained in vivo gene product delivery, but also for opening the way to therapy of other protein deficiencies for which gene therapy may be the only viable option. This review article describes, analyzes and compares the major animal models of GH gene therapy that have been developed in the last two decades. PMID- 16250892 TI - Immune response to herpes simplex virus and gamma134.5 deleted HSV vectors. AB - Herpes simplex virus (HSV) is a large DNA virus with unique properties that can be exploited for in vivo gene therapy. HSV is neurotropic, establishes latency, and has a large transgene capacity. These properties can be utilized in therapy of nervous system diseases. Wild-type HSV and the vectors derived from it induce both innate and acquired immune response. However, HSV is skillful in escaping the host response. It has evoked mechanisms including avoidance of antigen presentation on major histocompatibility (MHC) molecules, inhibition of host interferon response, impairment of the antibody and complement responses, and inhibition of apoptosis in infected cells. One of the molecules affecting the interferon response is ICP34.5, encoded by the so-called neurovirulence gene gamma(1)34.5. The mutants deleted of this gene are non-neurovirulent, having ca 3000-fold decreased ability to replicate in CNS. The HSV vectors based on the gamma(1)34.5 deletion mutants show efficacy against glioma and in other cancer therapies. These mutants provide an interesting platform for developing safe and efficient gene delivery for numerous neurological diseases or brain tumors. The immune response evoked by the HSV vector is central in determining the spread and persistence of the vector, and its transgene expression, and in controlling the innate and adaptive immune response against effective spread of the vector. These questions are key issues of herpesviral gene therapy and cancer therapy at the moment. This review describes the involvement of immune response in HSV infection and in gamma(1)34.5 deletion HSV-based virotherapy. We discuss the challenge of developing vectors with desired immune response benefiting the therapy and maintaining the efficiency. PMID- 16250893 TI - Transcription of the sarcoplasmic/endoplasmic reticulum Ca2+-ATPase type 3 gene, ATP2A3, is regulated by the calcineurin/NFAT pathway in endothelial cells. AB - Histamine, known to induce Ca2+ oscillations in endothelial cells, was used to alter Ca2+ cycling. Treatment of HUVEC (human umbilical-vein endothelial cell) derived EA.hy926 cells with histamine for 1-3 days increased the levels of SERCA (sarcoplasmic/endoplasmic reticulum Ca2+-ATPase) 3, but not of SERCA 2b, transcripts and proteins. Promoter-reporter gene assays demonstrated that this increase in expression was due to activation of SERCA 3 gene transcription. The effect of histamine was abolished by mepyramine, but not by cimetidine, indicating that the H1 receptor, but not the H2 receptor, was involved. The histamine-induced up-regulation of SERCA 3 was abolished by cyclosporin A and by VIVIT, a peptide that prevents calcineurin and NFAT (nuclear factor of activated T-cells) from interacting, indicating involvement of the calcineurin/NFAT pathway. Histamine also induced the nuclear translocation of NFAT. NFAT did not directly bind to the SERCA 3 promoter, but activated Ets-1 (E twenty-six-1), which drives the expression of the SERCA 3 gene. Finally, cells treated with histamine and loaded with fura 2 exhibited an improved capacity in eliminating high cytosolic Ca2+ concentrations, in accordance with an increase in activity of a low-affinity Ca2+-ATPase, like SERCA 3. Thus chronic treatment of endothelial cells with histamine up-regulates SERCA 3 transcription. The effect of histamine is mediated by the H1R (histamine 1 receptor) and involves activation of the calcineurin/NFAT pathway. By increasing the rate of Ca2+ sequestration, up regulation of SERCA 3 counteracts the cytosolic increase in Ca2+ concentration. PMID- 16250894 TI - Orchestra for assembly and fate of polyubiquitin chains. AB - Selective protein degradation by the 26 S proteasome usually requires a polyubiquitin chain attached to the protein substrate by three classes of enzymes: a ubiquitin-activating enzyme (E1), a ubiquitin-conjugating enzyme (E2), and a ubiquitin ligase (E3). This reaction can produce different polyubiquitin chains that, depending on size and linkage type, can provide distinct intracellular signals. Interestingly, polyubiquitination is sometimes regulated by additional conjugation factors, called E4s (polyubiquitin chain conjugation factors). Yeast UFD2 (ubiquitin fusion degradation protein-2), the first E4 to be described, binds to the ubiquitin moieties of preformed conjugates and catalyses ubiquitin-chain elongation together with E1, E2, and E3. Recent studies have illustrated that the E4 enzyme UFD2 co-operates with an orchestra of ubiquitin binding factors in an escort pathway to transfer and deliver polyubiquitinated substrates to the 26 S proteasome. Here we propose a model in which E4-dependent polyubiquitination pathways are modulated by different ubiquitin-binding proteins, using ataxin-3 as an example. PMID- 16250895 TI - E3 ubiquitin ligases. AB - The selectivity of the ubiquitin-26 S proteasome system (UPS) for a particular substrate protein relies on the interaction between a ubiquitin-conjugating enzyme (E2, of which a cell contains relatively few) and a ubiquitin-protein ligase (E3, of which there are possibly hundreds). Post-translational modifications of the protein substrate, such as phosphorylation or hydroxylation, are often required prior to its selection. In this way, the precise spatio temporal targeting and degradation of a given substrate can be achieved. The E3s are a large, diverse group of proteins, characterized by one of several defining motifs. These include a HECT (homologous to E6-associated protein C-terminus), RING (really interesting new gene) or U-box (a modified RING motif without the full complement of Zn2+-binding ligands) domain. Whereas HECT E3s have a direct role in catalysis during ubiquitination, RING and U-box E3s facilitate protein ubiquitination. These latter two E3 types act as adaptor-like molecules. They bring an E2 and a substrate into sufficiently close proximity to promote the substrate's ubiquitination. Although many RING-type E3s, such as MDM2 (murine double minute clone 2 oncoprotein) and c-Cbl, can apparently act alone, others are found as components of much larger multi-protein complexes, such as the anaphase-promoting complex. Taken together, these multifaceted properties and interactions enable E3s to provide a powerful, and specific, mechanism for protein clearance within all cells of eukaryotic organisms. The importance of E3s is highlighted by the number of normal cellular processes they regulate, and the number of diseases associated with their loss of function or inappropriate targeting. PMID- 16250896 TI - Proteasomes. AB - The major enzyme system catalysing the degradation of intracellular proteins is the proteasome system. A central inner chamber of the cylinder-shaped 20 S proteasome contains the active site, formed by N-terminal threonine residues. The 20 S proteasomes are extremely inefficient in degrading folded protein substrates and therefore one or two multisubunit 19 S regulatory particles bind to one or both ends of the 20 S proteasome cylinder, forming 26 S and 30 S proteasomes respectively. These regulatory complexes are able to bind proteins marked as proteasome substrates by prior conjugation with polyubiquitin chains, and initiate their unfolding and translocation into the proteolytic chamber of the 20 S proteasome, where they are broken down into peptides of 3-25 amino acids. The polyubiquitin tag is removed from the substrate protein by the deubiquitinating activity of the 19 S regulator complex. Under conditions of an intensified immune response, many eukaryotic cells adapt by replacing standard 20 S proteasomes with immuno-proteasomes and/or generating the proteasome activator complex, PA28. Both of these adaptations change the protein-breakdown process for optimized generation of antigenic peptide epitopes that are presented by the class I MHCs. Hybrid proteasomes (19 S regulator-20 S proteasome-PA28) may have a special function during the immune response. The functions of other proteasome accessory complexes, such as PA200 and PI31 are still under investigation. PMID- 16250897 TI - Ubiquitin-binding proteins: similar, but different. AB - Covalent modification of proteins with ubiquitin is a common regulatory mechanism in eukaryotic cells. Typically, ubiquitinated proteins are targeted for degradation by the 26 S proteasome. However, more recently the ubiquitin signal has also been connected with many other cell processes, including endocytosis, vesicle fusion, DNA repair and transcriptional silencing. Hence ubiquitination may be comparable with phosphorylation in its importance as an intracellular switch, controlling various signal-transduction pathways. Similar to the regulation of the extent of phosphorylation by kinases and phosphatases, specific sets of ubiquitinating/deubiquitinating enzymes control the degree of ubiquitination. A large number of ubiquitin-binding proteins act at different steps in the downstream pathways, followed by the ubiquitinated protein. Different families of ubiquitin-binding proteins have been described. UBA (ubiquitin-associated) domain-containing proteins is the largest family and includes members involved in different cell processes. The smaller groups of UIM (ubiquitin-interacting motif), GAT [GGA (Golgi-associated gamma-adaptin homologous) and Tom1 (target of Myb 1)], CUE (coupling of ubiquitin conjugation to endoplasmic reticulum degradation), UEV [ubiquitin E2 (ubiquitin-conjugating enzyme) variant] and NZF (nuclear protein localization gene 4 zinc finger) domain containing proteins appear to have more specialized functions. Here we discuss functional and structural properties of ubiquitin-binding proteins. PMID- 16250898 TI - Ubiquitin and control of transcription. AB - Eukaryotic transcription is one of the most complex cellular processes and constitutes the first step in protein synthesis. Ubiquitination and subsequent degradation by the 26 S proteasome, on the other hand, represents the final chapter in the life of a protein. Intriguingly, ubiquitin and the ubiquitin- proteasome system play vital roles in the regulation of transcription. Ubiquitin has dual modus operandi: firstly, ubiquitin functions via the 26 S proteasome--it is tagged to components of the transcription machinery, marking them for degradation via the proteasome, which results in the proper exchange of complexes during transcription and the prompt removal of activators after each round of transcription; and secondly, ubiquitin can function independently of the proteasome--histone ubiquitination results in heterochromatin relaxation and assembly of transcription complexes on the promoter, and ubiquitination of transcription factors enhances their transcriptional-activation function. Although ubiquitin and the ubiquitin-proteasome system were initially perceived as a graveyard for proteins, recent advances in molecular biological techniques have redefined their role as a regulatory system that influences the fate of many cellular processes, such as apoptosis, transcription and cell cycle progression. PMID- 16250899 TI - Ubiquitin and endocytic protein sorting. AB - Ubiquitin plays a fundamental role not only in proteasome-mediated protein degradation but also in the targeting of membrane proteins for degradation inside the lysosome. Ubiquitination provides a key signal for endosomal sorting of membrane proteins into the MVB (multi-vesicular body), which delivers its cargo to the proteolytic interior of the lysosome. Attachment of single ubiquitin molecules, rather than ubiquitin chains, to one or multiple lysines of the cytoplasmic domains of many growth factor receptors, ion channels and other membrane transporters is sufficient to target these proteins to a complex sorting apparatus on the endosome. This machinery selects ubiquitinated proteins for lysosomal sorting through consecutive interactions with a variety of ubiquitin binding domains. The major ubiquitin ligase (E3) responsible for ubiquitination in this pathway in yeast is the HECT [homologous to E6-AP (E6-associated protein) C-terminus]-ligase, Rsp5, whereas in mammalian cells the RING (really interesting new gene)-ligase Cbl has been implicated in the down-regulation of several RTKs (receptor tyrosine kinases). Ubiquitinated receptors can be rescued from degradation by the activity of DUBs (deubiquitinating enzymes), which may provide a proofreading mechanism that enhances the fidelity of this sorting and degradation process. DUBs also allow for recycling of the ubiquitin moieties from proteins prior to their final commitment to the MVB and lysosome interior. PMID- 16250900 TI - The role of the ubiquitin-proteasome system in ER quality control. AB - Misfolded endoplasmic reticulum (ER) proteins are eliminated by the retrotranslocation pathway in eukaryotes, which is an important physiological adaptation to ER stress. This pathway can be hijacked by certain viruses to destroy folded cellular proteins, such as MHC class I heavy chain. Recent studies have highlighted the importance of the ubiquitin-proteasome system (UPS) in this process. PMID- 16250901 TI - Fluorescent reporters for the ubiquitin-proteasome system. AB - Regulated turnover of proteins in the cytosol and nucleus of eukaryotic cells is primarily performed by the ubiquitin-proteasome system (UPS). The UPS is involved in many essential cellular processes. Alterations in this proteolytic system are associated with a variety of human pathologies, such as neurodegenerative diseases, cancer, immunological disorders and inflammation. The precise role of the UPS in the pathophysiology of these diseases, however, remains poorly understood. Detection of UPS aberrations has been a major challenge because of the complexity of the system. Most studies focus on various aspects of the UPS, such as substrate recognition, ubiquitination, deubiquitination or proteasome activity, and do not provide a complete picture of the UPS as an integral system. To monitor the efficacy of the UPS, a number of reporter substrates have been developed based on fluorescent proteins, such as the green fluorescent protein and its spectral variants. These fluorescent UPS reporters contain specific degradation signals that target them with high efficiency and accuracy for proteasomal degradation. Several studies have shown that these reporters can probe the functionality of the UPS in cellular and animal models and provide us with important information on the status of the UPS under various conditions. Moreover, these reporters can aid the identification and development of novel anti-cancer and anti-inflammatory drugs based on UPS inhibition. PMID- 16250902 TI - Transgenic manipulation of the ubiquitin-proteasome system. AB - The transgenic approach has been used in model systems from yeast to mammals to address basic research questions, and to achieve agricultural, pharmaceutical or industrial objectives. In basic research, transgenic organisms have generated novel observations that could not have been obtained otherwise. This chapter concentrates on the use of transgenics in deciphering the operation of the UPS (ubiquitin-proteasome system) in the yeast, plant, nematode, fly, and mouse model systems, and will touch on ways in which transgenic manipulation of the UPS has been exploited for agricultural, pharmaceutical, and industrial applications. PMID- 16250903 TI - Targeting of host-cell ubiquitin pathways by viruses. AB - The ability of viruses to co-opt cell signalling pathways has, over millions of years of co-evolution, come to pervade nearly every facet of cellular functions. Recognition of the extent to which the ubiquitin-proteasome system can be directed or subverted by viruses is relatively recent. Viral products interact with, and adjust, the ubiquitin-proteasome machinery precisely and at many levels, and they do so at distinct stages of viral life-cycles. The implications for both cells and viruses are fundamental, and understanding viral strategies in this context opens up fascinating new areas for research that span from basic cell biology to therapeutic interventions against both viruses and malignancies. PMID- 16250904 TI - The ubiquitin-proteasome system and neurodegenerative disorders. AB - As in all other mammalian tissues, the UPS (ubiquitin-proteasome system) is fundamental to normal brain function. A consistent feature of the major human neurodegenerative disorders is the accumulation of disease-related proteins, in non-native conformations, as protein aggregates within neurons or glial cells. Often the proteins in these aggregates are post-translationally conjugated with ubiquitin, suggesting a possible link between pathological protein-aggregation events in the nervous system and dysfunction of the UPS. Genetic evidence clearly demonstrates that disruption of ubiquitin-mediated processes can lead to neurodegeneration; however, the relationship between the UPS and idiopathic neurodegenerative disorders is less clear. In the latter cases, although a number of different mechanisms could potentially contribute to dysfunction of the UPS and promote the neurodegenerative process, whether UPS dysfunction is causally related to disease pathogenesis, or alternatively arises as a result of the pathological state, and indeed whether ubiquitinated inclusions are harmful or beneficial to cells, remains to be clarified. PMID- 16250905 TI - The ubiquitin-proteasome system and skeletal muscle wasting. AB - The ubiquitin-proteasome system (UPS) is believed to degrade the major contractile skeletal muscle proteins and plays a major role in muscle wasting. Different and multiple events in the ubiquitination, deubiquitination and proteolytic machineries are responsible for the activation of the system and subsequent muscle wasting. However, other proteolytic enzymes act upstream (possibly m-calpain, cathepsin L, and/or caspase 3) and downstream (tripeptidyl peptidase II and aminopeptidases) of the UPS, for the complete breakdown of the myofibrillar proteins into free amino acids. Recent studies have identified a few critical proteins that seem necessary for muscle wasting {i.e. the MAFbx (muscle atrophy F-box protein, also called atrogin-1) and MuRF-1 [muscle-specific RING (really interesting new gene) finger 1] ubiquitin-protein ligases}. The characterization of their signalling pathways is leading to new pharmacological approaches that can be useful to block or partially prevent muscle wasting in human patients. PMID- 16250906 TI - The ubiquitin-proteasome system and cancer. AB - The ubiquitin proteasome system (UPS) has emerged from obscurity to be seen as a major player in all regulatory processes in the cell. The concentrations of key proteins in diverse regulatory pathways are controlled by post-translational ubiquitination and degradation by the 26 S proteasome. These regulatory cascades include growth-factor-controlled signal-transduction pathways and multiple points in the cell cycle. The cell cycle is orchestrated by a combination of cyclin dependent kinases, kinase inhibitors and protein phosphorylation, together with the timely and specific degradation of cyclins and kinase inhibitors at critical points in the cell cycle by the UPS. These processes provide the irreversibility needed for movement of the cycle through gap 1 (G1), DNA synthesis (S), gap 2 (G2) and mitosis (M). The molecular events include cell-size control, DNA replication, DNA repair, chromosomal rearrangements and cell division. It is doubtful whether these events could be achieved without the temporally and spatially regulated combination of protein phosphorylation and ubiquitin dependent degradation of key cell-cycle regulatory proteins. The oncogenic transformation of cells is a multistep process that can be triggered by mutation of genes for proteins involved in regulatory processes from the cell surface to the nucleus. Since the UPS has critical functions at all these levels of control, it is to be expected that UPS activities will be central to cell transformation and cancer progression. PMID- 16250907 TI - Proteasome inhibitors as therapeutics. AB - The ubiquitin-proteasome pathway is a principle intracellular mechanism for controlled protein degradation and has recently emerged as an attractive target for anticancer therapies, because of the pleiotropic cell-cycle regulators and modulators of apoptosis that are controlled by proteasome function. In this chapter, we review the current state of the field of proteasome inhibitors and their prototypic member, bortezomib, which was recently approved by the U.S. Food and Drug Administration for the treatment of advanced multiple myeloma. Particular emphasis is placed on the pre-clinical research data that became the basis for eventual clinical applications of proteasome inhibitors, an overview of the clinical development of this exciting drug class in multiple myeloma, and a appraisal of possible uses in other haematological malignancies, such non Hodgkin's lymphomas. PMID- 16250908 TI - Towards a standardization of thrombin generation assessment: the influence of tissue factor, platelets and phospholipids concentration on the normal values of Thrombogram-Thrombinoscope assay. AB - BACKGROUND: Thrombin generation assay was developed several years ago to mimic physiological coagulation mechanisms but it had important limitations. Thrombogram-Thrombinoscope assay using a fluorogenic substrate, allows obtaining thrombin generation curves in non-defibrinated platelet rich plasma (PRP) in a fully automated manner. METHODS: We standardised the methodology of Thrombogram Thrombinoscope and we evaluated the precision of thrombin generation parameters (lag-time, maximum concentration of thrombin [Cmax], time required to reach Cmax [Tmax] and endogenous thrombin potential ETP) using different concentrations of recombinant human tissue factor, platelets or phospholipids. Normal values of thrombin generation assay were established in optimal experimental conditions. RESULTS: In the presence of low TF concentrations (final dilution of thromboplastin in plasma: 1/1000-1/2000) the Thrombogram assay showed intra-assay and inter-assay coefficients of variation lower than 9%. Thrombin generation parameters showed an important inter-individual variability and the coefficients of variation ranged from 18% to 50%. In PRP the lag-time, Cmax and Tmax but not the ETP, were influenced by TF concentration. Thrombin generation parameters were not influenced by variations of platelet concentration from 50 x 10(9)/l to 400 x 10(9)/l. The addition of synthetic procoagulant phospholipids in PPP strongly influenced all the parameters of thrombogram. For all the parameters of thrombogram a threshold effect was observed in the presence of phospholipid concentrations equal or higher to 4 microM. In frozen-thawed PRP the lag-time and the Tmax were significantly reduced and the Cmax was increased compared to the fresh PRP, but the ETP, the intra assay and the inter-assay coefficients of variation were similar in both test-systems. CONCLUSION: Thrombogram Thrombinoscope assay performed in fresh or in frozen-thawed PRP has an acceptable precision, with low inter-assay and intra-assay coefficient of variations. The concentration of TF is determinant for the normal values of the studied parameters of thrombin generation. When the assay is performed in PPP, thrombin generation parameters are influenced by the concentration of procoagulant synthetic phospholipids. The optimal experimental conditions were obtained in the presence of 1/1000 final dilution of thromboplastin, a platelet count higher than 50 x 10(9)/l and a synthetic phospholipid concentration higher than 4 microM. PMID- 16250910 TI - A randomised controlled trial to assess the effectiveness of offering study results as an incentive to increase response rates to postal questionnaires [ISRCTN26118436]. AB - BACKGROUND: Postal questionnaires are widely used to collect outcome data on participants. However, a poor response to questionnaires will reduce the statistical power of the study and may introduce bias. A meta analysis of ten trials offering study results, largely in the fields of education and marketing, was shown to be ineffective, with the odds ratio for response with offering research findings is 0.92 (95% CI 0.75 to 1.11). However uncertainty still exists as it is uncertain whether results from such trials can be extrapolated to that of a health care setting. The aim of this study was to assess whether offering participants study results increases the response rates to postal questionnaires. METHODS: 1038 women aged over 70 years were remotely randomised by computer in a 3:1 ratio. 250 participants did not receive the offer of knowing the results of the trial and 788 participants were offered the results of the trial in a postal questionnaire. The main outcome measure was response rate. Chi square test was used to evaluate the overall differences in response rate between the two groups. An adjusted analysis, adjusting for whether the participant was taking calcium and age was also undertaken. RESULTS: The response rates were not significantly different Odds Ratio 0.88 (95% confidence intervals 0.48 to 1.63) p = 0.69. CONCLUSION: Offering study results to women living in the community aged over 70 does not increase response rates to postal questionnaires. Although researchers have an ethical obligation to offer participants study results, since 10% of women did not wish to receive the results, investigators should give participants the option to opt out of receiving the study's results. PMID- 16250909 TI - Multiple P2Y receptors couple to calcium-dependent, chloride channels in smooth muscle cells of the rat pulmonary artery. AB - BACKGROUND: Uridine 5'-triphosphate (UTP) and uridine 5'-diphosphate (UDP) act via P2Y receptors to evoke contraction of rat pulmonary arteries, whilst adenosine 5'-triphosphate (ATP) acts via P2X and P2Y receptors. Pharmacological characterisation of these receptors in intact arteries is complicated by release and extracellular metabolism of nucleotides, so the aim of this study was to characterise the P2Y receptors under conditions that minimise these problems. METHODS: The perforated-patch clamp technique was used to record the Ca2+ dependent, Cl- current (I(Cl,Ca)) activated by P2Y receptor agonists in acutely dissociated smooth muscle cells of rat small (SPA) and large (LPA) intrapulmonary arteries, held at -50 mV. Contractions to ATP were measured in isolated muscle rings. Data were compared by Student's t test or one way ANOVA. RESULTS: ATP, UTP and UDP (10(-4) M) evoked oscillating, inward currents (peak = 13-727 pA) in 71 93% of cells. The first current was usually the largest and in the SPA the response to ATP was significantly greater than those to UTP or UDP (P < 0.05). Subsequent currents tended to decrease in amplitude, with a variable time-course, to a level that was significantly smaller for ATP (P < 0.05), UTP (P < 0.001) and UDP (P < 0.05) in the SPA. The frequency of oscillations was similar for each agonist (mean approximately to 6-11.min(-1)) and changed little during agonist application. The non-selective P2 receptor antagonist suramin (10(-4) M) abolished currents evoked by ATP in SPA (n = 4) and LPA (n = 4), but pyridoxalphosphate-6-azophenyl-2',4'-disulphonic acid (PPADS) (10(-4) M), also a non-selective P2 antagonist, had no effect (n = 4, 5 respectively). Currents elicited by UTP (n = 37) or UDP (n = 14) were unaffected by either antagonist. Contractions of SPA evoked by ATP were partially inhibited by PPADS (n = 4) and abolished by suramin (n = 5). Both antagonists abolished the contractions in LPA. CONCLUSION: At least two P2Y subtypes couple to ICl,Ca in smooth muscle cells of rat SPA and LPA, with no apparent regional variation in their distribution. The suramin-sensitive, PPADS-resistant site activated by ATP most resembles the P2Y11 receptor. However, the suramin- and PPADS-insensitive receptor activated by UTP and UDP does not correspond to any of the known P2Y subtypes. These receptors likely play a significant role in nucleotide-induced vasoconstriction. PMID- 16250911 TI - Expression of thromboxane synthase, TBXAS1 and the thromboxane A2 receptor, TBXA2R, in human breast cancer. AB - BACKGROUND: Thromboxane synthase (TxS) metabolizes the cyclooxygenase product, prostaglandin H(2), into thromboxanes. Some of the thromboxanes are known to be biologically active on cancer cells. The aim of the study was to investigate the expression of thromboxane synthases, TBXAS1 and the thromboxane A2 receptor, TBXA2R in a cohort of human breast cancer patients and also to assess their potential clinical relevance. METHODS: Human breast tumour tissues (n = 120) and non-neoplastic mammary tissues (n = 32) were studied. Levels of TBXA2R and TBXAS1 transcripts were quantified using quantitative real-time RT-PCR analysis and correlated with clinical/pathological information including nodal status, grade, prognosis and long term survival (median follow-up period 120 months). RESULTS: Breast tumour tissue expressed higher levels of TBXA2R compared with normal mammary tissues, although the difference was not statistically significant (p = 0.09). There was no difference between tumour and normal tissues for TBXAS1. However, TBXA2R expression was significantly increased in grade 3 tumours(p = 0.006 vs grade 1), while TBXAS1 was significantly reduced in grade 3 tumours (p = 0.026 vs grade 1 tumours). A similar differential expression pattern was seen in tumours from patients with different prognosis, in that patients with predicted poor prognosis had higher, but not statistically different, levels of TBXA2R, and significantly lower levels of TBXAS1 (p = 0.008). Finally, Kaplan-Meier survival analysis has shown that patients with high levels of TBXA2R had significantly shorter disease free survival (103.8 (79.1-128.5) months) compared with those with low levels (123.7 (112.0-135.3)) months, p = 0.043. CONCLUSION: Thromboxane synthases are differentially expressed in human breast cancer. While TBXA2R is highly expressed in aggressive tumours and linked with poor prognosis, TBXAS1 is expressed at significantly low levels in high grade tumours and tumour patients with poor prognosis. TBXA2R thus has a significant prognostic value in clinical breast cancer. PMID- 16250912 TI - Influences of dietary soy isoflavones on metabolism but not nociception and stress hormone responses in ovariectomized female rats. AB - BACKGROUND: Isoflavones, the most abundant phytoestrogens in soy foods, are structurally similar to 17beta-estradiol. Few studies have examined the nociception and stress hormone responses after consumption of soy isoflavones. METHODS: In this study, ovariectomized (OVX) female Long-Evans rats were fed either an isoflavone-rich diet (Phyto-600) or an isoflavone-free diet (Phyto free). We examined the effects of soy isoflavones on metabolism by measuring body weights, food/water intake, adipose tissue weights as well as serum leptin levels. Also, circulating isoflavone levels were quantified. During chemically induced estrous, nociceptive thresholds were recorded. Then, the animals were subjected to a stressor and stress hormone levels were quantified. RESULTS: Body weights were significantly lower in Phyto-600 fed rats compared to Phyto-free values within one week and during long-term consumption of soy isoflavones. Correspondingly, Phyto-600 fed animals displayed significantly less adipose deposition and lower serum leptin levels than Phyto-free values. However, rats on the Phyto-600 diet displayed greater food/water intake compared to Phyto-free levels. No changes in thermal pain threshold or stress hormone levels (ACTH and corticosterone) were observed after activation of the hypothalamic-pituitary adrenal (HPA) stress axis. CONCLUSION: In summary, these data show that consumption of soy isoflavones 1) increases metabolism, demonstrated by significantly decreased body weights, adipose tissue deposition and leptin levels, but 2) does not alter nociception or stress hormone responses, as indexed by thermal pain threshold, serum corticosterone and ACTH levels in chemically induced estrous OVX rats. PMID- 16250913 TI - Dobutamine stress echocardiography in healthy adult male rats. AB - BACKGROUND: Dobutamine stress echocardiography is used to investigate a wide variety of heart diseases in humans. Dobutamine stress echocardiography has also been used in animal models of heart disease despite the facts that the normal response of healthy rat hearts to this type of pharmacological stress testing is unknown. This study was performed to assess this normal response. METHODS: 15 normal adult male Wistar rats were evaluated. Increasing doses of dobutamine were infused intravenously under continuous imaging of the heart by a 12 MHz ultrasound probe. RESULTS: Dobutamine stress echocardiography reduced gradually LV diastolic and systolic dimensions. Ejection fraction increased by a mean of +24% vs. baseline. Heart rate increased progressively without reaching a plateau. Changes in LV dimensions and ejection fraction reached a plateau after a mean of 4 minutes at a constant infusion rate. CONCLUSION: DSE can be easily performed in rats. The normal response is an increase in heart rate and ejection fraction and a decrease in LV dimensions. A plateau in echocardiographic measurements is obtained after 4 minutes of a constant infusion rate in most animals. PMID- 16250914 TI - Small bowel Gastrointestinal Stromal Tumors can physiologically alter gut motility before causing mechanical obstruction. AB - BACKGROUND: Gastro Intestinal Stromal Tumors (GISTs) are rare stromal neoplasms that represent the most common mesenchymal tumor of the G.I. tract, accounting for 5% of all sarcomas. Originating from interstitial cells of Cajal, which are regulators of gut peristalsis, they are preferentially located in the stomach and the small intestine and clinical presentation is variable, ranging from vague complaints to major G.I. bleeding. Surgical resection is the mainstay of treatment for patients with resectable GIST and 5-year survival ranges from 21% to 88% in different series depending on risk grading and completeness of surgical resection. Imatinib mesylate, a tyrosine kinase inhibitor, provides an encouraging option for treating high risk GISTs. CASE PRESENTATION: We report the case of a 62-year-old lady who had been diagnosed and being treated unsuccessfully for Irritable bowel syndrome for 11 years and eventually found to have an obstructing small bowel GIST. CONCLUSION: The symptoms from GIST may mimic those of irritable bowel syndrome. A physiological alteration in gut peristalsis resulting from neoplastic transformation of the interstitial cells of Cajal, is a hypothesis that could explain this presentation. An alternative diagnosis should be considered when treating patients with irritable bowel syndrome who fail to respond for a prolonged period. PMID- 16250915 TI - Chance mechanisms affecting the burden of metastases. AB - BACKGROUND: The burden of cancer metastases within an individual is commonly used to clinically characterize a tumor's biological behavior. Assessments like these implicitly assume that spurious effects can be discounted. Here the influence of chance on the burden of metastasis is studied to determine whether or not this assumption is valid. METHODS: Monte Carlo simulations were performed to estimate tumor burdens sustained by individuals with cancer, based upon empirically derived and validated models for the number and size distributions of metastases. Factors related to the intrinsic metastatic potential of tumors and their host microenvironments were kept constant, to more clearly demonstrate the contribution from chance. RESULTS: Under otherwise identical conditions, both the simulated numbers and the sizes of metastases were highly variable. Comparable individuals could sustain anywhere from no metastases to scores of metastases, and the sizes of the metastases ranged from microscopic to macroscopic. Despite the marked variability in the number and sizes of the metastases, their respective growth times were rather more narrowly distributed. In such situations multiple occult metastases could develop into fully overt lesions within a comparatively short time period. CONCLUSION: Chance can have a major effect on the burden of metastases. Random variability can be so great as to make individual assessments of tumor biology unreliable, yet constrained enough to lead to the apparently simultaneous appearance of multiple overt metastases. PMID- 16250916 TI - Determination of biological and physicochemical parameters of Artemia franciscana strains in hypersaline environments for aquaculture in the Colombian Caribbean. AB - BACKGROUND: Artemia (Crustacea, Anostraca), also known as brine shrimp, are typical inhabitants of extreme environments. These hypersaline environments vary considerably in their physicochemical composition, and even their climatic conditions and elevation. Several thalassohaline (marine) environments along the Colombian Caribbean coast were surveyed in order to contribute to the knowledge of brine shrimp biotopes in South America by determining some vital biological and physicochemical parameters for Artemia survival. Additionally, cyst quality tests, biometrical and essential fatty acids analysis were performed to evaluate the economic viability of some of these strains for the aquaculture industry. RESULTS: In addition to the three locations (Galerazamba, Manaure, and Pozos Colorados) reported in the literature three decades ago in the Colombian Caribbean, six new locations were registered (Salina Cero, Kangaru, Tayrona, Bahia Hondita, Warrego and Pusheo). All habitats sampled showed that chloride was the prevailing anion, as expected, because of their thalassohaline origin. There were significant differences in cyst diameter grouping strains in the following manner according to this parameter: 1) San Francisco Bay (SFB-Control, USA), 2) Galerazamba and Tayrona, 3) Kangaru, 4) Manaure, and 5) Salina Cero and Pozos Colorados. Chorion thickness values were smaller in Tayrona, followed by Salina Cero, Galerazamba, Manaure, SFB, Kangaru and Pozos Colorados. There were significant differences in naupliar size, grouping strains as follows (smallest to largest): 1) Galerazamba, 2) Manaure, 3) SFB, Kangaru, and Salina Cero, 4) Pozos Colorados, and 5) Tayrona. Overall, cyst quality analysis conducted on samples from Manaure, Galerazamba, and Salina Cero revealed that all sites exhibited a relatively high number of cysts.g-1. Essential fatty acids (EFA) analysis performed on nauplii from cyst samples from Manaure, Galerazamba, Salina Cero and Tayrona revealed that cysts from all sites exhibited high arachidonic acid:20:4(n-6) (ArA) and eicosapentaenoic acid: 20:5(n-3) (EPA) levels comparable to the control sample (SFB). In contrast, most cysts collected (including SFB) at different locations, and during different months, presented low docosahexaenoic acid: 22:6(n-3) (DHA) levels (Manaure was the only exception with high DHA levels). Some variations in EPA and ArA levels were observed in all sites, contrasting with the much lower DHA levels which remained constant for all locations, except for Manaure which exhibited variable DHA levels. DHA/EPA ratio was overall very low for all sites compared to SFB cysts. All strains had a low DHA/ArA, but a high EPA/ArA ratio, including the control. CONCLUSION: The Colombian A. franciscana habitats analyzed were determined to be thalassohaline, and suitable for A. franciscana development. EFA profiles demonstrated that Tayrona, Galerazamba, Manaure and Salina Cero strains are suitable food for marine fish and crustacean culture because of their high EPA/ArA ratio, but might have to be fortified with DHA rich emulsions depending on the nutritional requirements of the species to be cultured, because of their overall low DHA content. The relatively small nauplii are appropriate for marine larvaeculture. In contrast, the strains from Tayrona, Kangaru, Salina Cero, and Pozos Colorados may be of use but limited to Artemia small biomass production quantities, because of the small surface area of their respective locations; Artemia could be exploited at these locations for local aquaculture applications. In general, cyst quality evaluation for Manaure, Salina Cero and Galerazamba cysts revealed that cysts from these three locations could improve their quality by concentrating efforts on cyst processing techniques. Finally, most locations had great A. franciscana production potential and require different degrees of water quality and/or infrastructure management. PMID- 16250917 TI - HDACs and the senescent phenotype of WI-38 cells. AB - BACKGROUND: Normal cells possess a limited proliferative life span after which they enter a state of irreversible growth arrest. This process, known as replicative senescence, is accompanied by changes in gene expression that give rise to a variety of senescence-associated phenotypes. It has been suggested that these gene expression changes result in part from alterations in the histone acetylation machinery. Here we examine the influence of HDAC inhibitors on the expression of senescent markers in pre- and post-senescent WI-38 cells. RESULTS: Pre- and post-senescent WI-38 cells were treated with the HDAC inhibitors butyrate or trichostatin A (TSA). Following HDAC inhibitor treatment, pre senescent cells increased p21WAF1 and beta-galactosidase expression, assumed a flattened senescence-associated morphology, and maintained a lower level of proteasome activity. These alterations also occurred during normal replicative senescence of WI-38 cells, but were not accentuated further by HDAC inhibitors. We also found that HDAC1 levels decline during normal replicative senescence. CONCLUSION: Our findings indicate that HDACs impact numerous phenotypic changes associated with cellular senescence. Reduced HDAC1 expression levels in senescent cells may be an important event in mediating the transition to a senescent phenotype. PMID- 16250918 TI - Axial stent strut angle influences wall shear stress after stent implantation: analysis using 3D computational fluid dynamics models of stent foreshortening. AB - INTRODUCTION: The success of vascular stents in the restoration of blood flow is limited by restenosis. Recent data generated from computational fluid dynamics (CFD) models suggest that the vascular geometry created by an implanted stent causes local alterations in wall shear stress (WSS) that are associated with neointimal hyperplasia (NH). Foreshortening is a potential limitation of stent design that may affect stent performance and the rate of restenosis. The angle created between axially aligned stent struts and the principal direction of blood flow varies with the degree to which the stent foreshortens after implantation. METHODS: In the current investigation, we tested the hypothesis that stent foreshortening adversely influences the distribution of WSS and WSS gradients using time-dependent 3D CFD simulations of normal arteries based on canine coronary artery measurements of diameter and blood flow. WSS and WSS gradients were calculated using conventional techniques in ideal (16 mm) and progressively foreshortened (14 and 12 mm) stented computational vessels. RESULTS: Stent foreshortening increased the intrastrut area of the luminal surface exposed to low WSS and elevated spatial WSS gradients. Progressive degrees of stent foreshortening were also associated with strut misalignment relative to the direction of blood flow as indicated by analysis of near-wall velocity vectors. CONCLUSION: The current results suggest that foreshortening may predispose the stented vessel to a higher risk of neointimal hyperplasia. PMID- 16250921 TI - Complementary and alternative medicine in women's health: healing or hoax? PMID- 16250920 TI - Easy and rapid method for the determination of gene expression in cumulus cells incubated for oocyte maturation. AB - BACKGROUND: The objectives of this study were to develop an easy and rapid method for measuring gene expression in a small number of cells by real-time PCR without RNA extraction and purification, and to use this method to determine more precisely IGF-I gene expression in the cumulus cells surrounding oocytes. METHODS: First, after small numbers of cumulus cells were lysed in cell lysis buffer, they were digested with various concentrations of DNase I for different periods at 37 degrees C to determine the optimal conditions for digestion of genomic DNA in the lysate. Since nonspecific amplification was liable to occur when the non-purified RT product of the cell lysate was used for real-time PCR with the given primers, the optimal conditions for Mg2+ and annealing temperature were well investigated. Further, to create the same conditions as in the actual sample reaction for measurement by real-time PCR, RT-minus product was added to the reaction mixture of the standard curve, and then the amplification efficiency was assessed. Next, IGF-I gene expression in cumulus cells collected from cumulus oocyte complexes (COCs) every 4 h during maturation was determined using the developed method. RESULTS: The optimal conditions for measuring gene expression using the cell lysate from a small number of cells were as follows: incubation of the cell lysate with 0.16 U/microL DNase I with 10 U/microL for 30 min, an Mg concentration of 1.5 mM for amplification of target gene by real-time PCR using RT-product of the cell lysate. When the RT-minus products added to the reaction mixture for the standard curve, which was prepared for purified 18SrRNA plasmid, the PCR efficiency was similar between the sample and the standard. The IGF-I gene expression in the cumulus cells was elevated up through the first 8 h of the culture and then declined gradually by the end of maturation, with the maximal gene expression (778-fold) seen at 8 h. CONCLUSION: It can be concluded that the method developed here, in which equivalent to cumulus cells collected from 0.03 0.075 COCs were employed per reaction, permits rapid and easy determination of target gene expression in a limited number of cells using real-time PCR without RNA extraction. PMID- 16250919 TI - Barriers to successful care for chronic kidney disease. AB - BACKGROUND: The National Kidney Foundation has formulated clinical practice guidelines for patients with chronic kidney disease (K/DOQI). However, little is know about how many patients actually achieve these goals in a dedicated clinic for chronic kidney disease. METHODS: We performed a cross-sectional analysis of 198 patients with an estimated glomerular filtration rate of less than 30 ml/min/1.73 m2 and determined whether K/DOQI goals were met for calcium, phosphate, calcium-phosphate product, parathyroid hormone, albumin, bicarbonate, hemoglobin, lipids, and blood pressure. RESULTS: We found that only a small number of patients achieved K/DOQI targets. Recent referral to the nephrologist, failure to attend scheduled clinic appointments, African American ethnicity, diabetes, and advanced renal failure were significant predictors of low achievement of K/DOQI goals. CONCLUSION: We conclude that raising awareness of chronic kidney disease and K/DOQI goals among primary care providers, early referral to a nephrologist, the exploration of socioeconomic barriers and cultural differences, and both patient and physician education are critical to improve CKD care in patients with Stage 4 and 5 CKD. PMID- 16250922 TI - Antepartum bleeding of unknown origin in the second half of pregnancy: a review. AB - The objective of this study was to determine the prevalence, adverse pregnancy complications, and optimal management of pregnancies complicated by bleeding in the second half of pregnancy of an unknown origin (ABUO). A MEDLINE search from 1966 through November 2004 using the search terms "antepartum hemorrhage" or "hemorrhage" or "uterine hemorrhage" and "pregnancy complications" and "cardiovascular complications" and "second trimester pregnancy" or "third trimester pregnancy" was undertaken. The inclusion criteria focused on bleeding not resulting from placenta previa or abruption or to any known cause. The MEDLINE search provided 24 abstracts for review with 9 studies meeting the inclusion criteria The prevalence of ABUO was 2%. The likelihood of antepartum hemorrhage and delivery before 37 weeks was significant with an odds ratio (OR) of 3.17 and 95% confidence interval (CI) of 2.76-3.64. The risk of intrauterine fetal demise was significantly increased in women with ABUO (OR, 2.09; 95% CI, 1.43-3.06). The association between ABUO and fetal anomalies was increased with an OR 1.42 (95% CI, 1.07-1.87). Only one study with a small sample size (N = 48) compared the outcomes of women using Doppler studies of the umbilical and uterine arteries and biophysical profiles. No differences were observed in the women undergoing antenatal testing and the women not undergoing antenatal testing. The prevalence of ABUO is 2%. Preterm delivery, stillbirth, and fetal anomalies appear to be increased in these pregnancies. Antenatal testing may be of limited value in their management. TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians Learning Objectives: After completion of this article, the reader should be able to explain the prevalence of antepartum bleeding of unknown origin (ABUO) in confronting a patient with ABUO, summarize the types and frequency of adverse pregnancy outcomes in ABUO, and recall the limited usefulness of antenatal testing in patients with ABUO. PMID- 16250923 TI - Nonhormonal drugs for contraception in men: a systematic review. AB - Nonhormonal drugs for contraception in men may have advantages over hormonal methods. The nonhormonal methods can have more rapid onset and less interference with androgen-dependent functions. This systematic review summarizes the clinical studies evaluating nonhormonal drugs administered to men for contraception. Relevant clinical results were found for gossypol, which is derived from the cotton plant, and for extracts of Tripterygium, a plant used in Chinese traditional medicine. Randomized, controlled trials were available on the efficacy of gossypol and on the effect of gossypol on potassium levels. Gossypol had problems with low efficacy and toxicity. For Tripterygium, 2 observational studies described men who were treated for rheumatoid arthritis. Although sperm density was lower among those taking Tripterygium, later reports indicated some toxicity. Nonclinical research continues on isolates of Tripterygium. No clinical studies for contraception in men were found for nonhormonal vaccines or neem, which is also a plant used for medicinal purposes. Clinical trials studied injecting styrene maleic anhydride into the vas deferens, but no comparative data were provided. At this time, no safe and effective nonhormonal drug is available for contraception in men. TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians. LEARNING OBJECTIVES: After completion of this article, the reader should be able to state that the number of studies concerning the use of nonhormonal drugs for male contraception are very limited, point out that the two nonhormonal drugs used to a small degree have varying results and serious side effects, and recall that there are limited clinical studies on use of vas deferens injections and vaccines in humans. PMID- 16250924 TI - Rectocele repair using biomaterial augmentation: current documentation and clinical experience. AB - Although the etiology of rectocele remains debated, surgical innovations are currently promoted to improve anatomic outcome while avoiding dyspareunia and alleviating rectal emptying difficulties following rectocele surgery. Use of biomaterials in rectocele repair has become widespread in a short time, but the clinical documentation of their effectiveness and complications is limited. Medline and the Cochrane database were searched electronically from 1964 to May 2005 using the Pubmed and Ovid search engines. All English language publications including any of the search terms "rectocele," "implant," "mesh," "biomaterial," "prolapse," "synthetical," "pelvic floor," "biological," and "compatibility" were reviewed. This review outlines the basic principles for use of biomaterials in pelvic reconstructive surgery and provides a condensation of peer-reviewed articles describing clinical use of biomaterials in rectocele surgery. Historical and new concepts in rectocele surgery are discussed. Factors of importance for human in vivo biomaterial compatibility are presented together with current knowledge from clinical studies. Potential risks and problems associated with the use of biomaterials in rectocele and pelvic reconstructive surgery in general are described. Although use of biomaterials in rectocele and other pelvic organ prolapse surgery offers exciting possibilities, it raises treatment costs and may be associated with unknown and potentially severe complications at short and long term. Clinical benefits are currently unknown and need to be proven in clinical studies. TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians TARGET AUDIENCE: After completion of this article, the reader should be able to explain that the objective of surgical treatment is to improve anatomic outcome and alleviate rectal emptying difficulties, describe the efficacy of biomaterials in rectocele repair, and summarize the potential risks and problems associated with use of biomaterials in rectocele and pelvic reconstructive surgery. PMID- 16250925 TI - Uterine arteriovenous malformations: a review of the current literature. AB - Uterine arteriovenous malformation (AVM) is a rare but potentially life threatening condition. AVMs often present with intractable bleeding and commonly are seen in association with pregnancy and uterine trauma. Ultrasound is the most common form of initial investigation, and computed tomography and magnetic resonance imaging are being used with greater frequency. Despite this, angiography remains the gold standard for diagnostic evaluation. Embolization has become a more acceptable form of treatment and allows more invasive forms of treatment, particularly hysterectomy, to be avoided. Numerous medical therapies have also been used in the management of patients with uterine AVM. Reports of successful pregnancies after diagnosis and treatment of a uterine AVM are still uncommon, but increasingly good outcomes are being reported after successful treatment of a confirmed uterine AVM. TARGET AUDIENCE: Obstetricians & Gynecologists, Family Physicians Learning Objectives: After completion of this article, the reader should be able to describe the many and varied clinical manifestations of a uterine arteriovenous malformation (AVM), summarize the best ways to manage an acute hemorrhage from an AVM, and identify the current best way to diagnose an AVM. PMID- 16250927 TI - Preclinical data with bortezomib in lung cancer. AB - More effective therapies are needed for non-small-cell lung cancer (NSCLC) and small-cell lung cancer (SCLC). Proteasome inhibitors are one class of molecularly targeted antineoplastic agents being investigated for these diseases. These agents block the activity of the 26S proteasome, which is responsible for the degradation of the vast majority of intracellular proteins and thus affect multiple signaling pathways within cells. Bortezomib is the first proteasome inhibitor to be evaluated in human studies and is approved for use in multiple myeloma. Bortezomib is now being investigated as a potential treatment for NSCLC and SCLC. Preclinical studies have shown that single-agent bortezomib causes growth inhibition and apoptosis in numerous NSCLC cell lines in vitro and has antitumor activity in vivo. Bortezomib affects the levels of several proteins known to be of significance in lung cancers. Studies of bortezomib in combination with other antitumor agents in vitro and in vivo demonstrate that these combination regimens can offer additive/synergistic effects compared with the single agents. Bortezomib has been investigated in combination with taxanes, gemcitabine, carboplatin, histone deactylase inhibitors, and other molecularly targeted agents in various NSCLC cell lines. The sequence of administration of the agents in preclinical combination regimens in vitro and in vivo has been shown to be of significance; further elucidation of the mechanism of efficacy of bortezomib in lung cancer is required. Numerous clinical studies have been carried out or are ongoing. Bortezomib has the potential to play a significant role in the future management of NSCLC and SCLC. PMID- 16250928 TI - Sequencing bortezomib with chemotherapy and targeted agents. AB - The 26S proteasome is a multicatalytic threonine protease complex that is responsible for intracellular protein turnover in eukaryotic cells. This complex degrades and processes proteins required for regulation of various cellular functions. Bortezomib is a novel proteasome inhibitor approved for therapy of multiple myeloma. Inhibition of ubiquitin-proteasome-mediated protein degradation by bortezomib leads to accumulation of its diverse substrates, including cyclins, transcriptional factors, tumor suppressor proteins, and protooncogenes. The sequelae of such profound perturbation of cellular function include cell cycle arrest and activation of apoptotic programs. As the development of this agent continues, there is interest in evaluating its interaction with other anticancer agents. This review provides an overview of selected interactions between bortezomib and other anticancer agents preclinically and in early clinical trials. PMID- 16250929 TI - Bortezomib-based combinations in the treatment of non-small-cell lung cancer. AB - Lung cancer is the most common cause of cancer-related death among men and women in the United States. Current trials are focusing on the integration of novel therapeutic agents into current non-small-cell lung cancer (NSCLC) treatment paradigms. Bortezomib, a small-molecule proteasome inhibitor, has single-agent activity in NSCLC and in combination with agents commonly used in NSCLC. This article will review the rationale and preclinical data supporting bortezomib combinations and the clinical trials with bortezomib alone and in combination in NSCLC to date. PMID- 16250930 TI - The potential role of bortezomib in combination with chemotherapy and radiation in non-small-cell lung cancer. AB - The combination of chemotherapy and radiation has been validated for the treatment of locally advanced non-small-cell lung cancer (NSCLC). However, the results are still unsatisfactory, and there is a need to improve current treatment. One approach is to use new agents that have the potential to enhance the efficacy of chemotherapy, radiation therapy (RT), or both. One potential target is the ubiquitin-proteasome pathway. This pathway plays an essential role in the degradation of most short- and long-lived intracellular proteins in eukaryotic cells and therefore regulating the cell cycle, neoplastic growth, and metastasis. Bortezomib is a selective 26S proteasome inhibitor that has been approved for the treatment of multiple myeloma. Bortezomib has demonstrated in vitro chemotherapy- and RT-sensitizing properties as well as single-agent activity in lung cancer. This article will review the rationale for the use of bortezomib as part of the chemotherapy/RT strategy for the treatment of NSCLC. PMID- 16250931 TI - Proteasome inhibition in small-cell lung cancer: preclinical rationale and clinical applications. AB - Small-cell lung cancer (SCLC) is a tobacco-related malignancy that usually presents in an extensive and therefore incurable stage. Although initially sensitive to platinum agent-based therapy, SCLC rapidly becomes refractory to chemotherapy, leading to disease recurrence and ultimately patient death. Treatment options following failure of first-line platinum agent-based therapy are limited. Small-cell lung cancer is characterized by molecular aberrancies such as overexpression of the antiapoptotic protein Bcl-2, which is regulated in part by the inhibitory IkappaB, a target of the ubiquitin-proteasome degradative pathway. Bortezomib is a proteasome inhibitor that can decrease Bcl-2 expression through diminished IkappaB degradation. Efforts to promote apoptosis in SCLC through the integration of bortezomib into therapy are under way. PMID- 16251009 TI - Mediterranean dietary components and body mass index in adults: the peel nutrition and heart health survey. AB - Diet is a lifestyle factor that contributes to the risk of overweight/obesity and cardiovascular disease. The objective of this study was to examine the hypothesis that a Mediterranean-type dietary pattern (M) is associated with healthy body weights in a large suburban municipality in Ontario. A random cross-sectional sample of 759 adults, 18 to 65 years of age, participated in a telephone survey, which included questions on the frequency of consumption of 60 food categories. Principal components analysis showed that food categories aggregated into six low order dietary factors and two high-order dietary patterns. The M pattern reflected higher consumption of fruits and vegetables, olive oil and garlic, and fish and shellfish. The non-M pattern reflected high fat/nutrient poor, meats and poultry, and foods high in added sugars. The M-score was inversely related to body mass index (BMI) (p = 0.027). After adjustment for gender, education, income and marital status, a higher M-score predicted a lower BMI in the 40 to 49 year age group. Heart health promotion strategies aimed at preventing adult obesity should emphasize components of a Mediterranean-type diet pattern. PMID- 16251010 TI - Screening mammography participation and invitational strategy: the Quebec Breast Cancer Screening Program, 1998-2000. AB - In the Quebec Breast Cancer Screening Program, a personalized letter signed by a regional program physician is sent to every woman in the province 50 to 69 years of age, inviting her to have a screening mammogram. A reminder letter is also frequently sent. The aim of this study was to evaluate the influence of this screening invitational strategy on rates of participation. The population studied was comprised of 684,028 women in Quebec aged 50-69. The baseline (expected) monthly mammography screening rate was estimated from the rate of screening mammograms recorded between the date a woman became eligible for screening and the mailing date of her personalized invitational letter; the observed monthly mammography screening rate was calculated after the mailing of the letter. Compared to baseline (expected) screening rates, observed rates were substantially increased (p<.05). The ratios of observed to expected rates were respectively 3.05 and 2.23 in the second and fourth months, respectively, after the letter mailing, coinciding with the mailing of the initial and reminder letters. In the twelve months after the mailing, the ratio of observed to expected rates was 1.68 (95% CI: 1.67-1.69). Twelve months following the mailing, 30 percent of the women who were letter recipients had undergone a screening mammography, compared to an expected cumulative probability of 20 percent for women not receiving a letter. The strength of this effect was similar to one seen in randomised controlled trials. PMID- 16251011 TI - An observational study of sun and heat protection during Canada Day outdoor celebration, 2003. AB - Attendance at summer outdoor mass gatherings may lead to heat- and sun-related illness. The purposes of this study were: (1) to estimate the proportion of people in attendance at the 2003 Canada Day celebration in the National Capital Region who used sun and heat protective items; (2) to identify factors associated with the utilization of these protective items; and (3) to provide research data to public outdoor event organizers when developing evidence-based plans for safer events. A naturalistic observational cross-sectional method was used to gather information at the 2003 Canada Day celebration in the National Capital Region on attendees' demographics, the sun and heat protective items they used and the protective resources available at the event sites. Of the 398 observed attendees, the proportion using any one of the protective items ranged from 3 percent (an open umbrella) to 51.5 percent (sunglasses). Females were more likely to use protective items more than males, and adults more likely than children. Planners of public outdoor events should consider the factors that influence the utilization of sun and heat protective behaviours and the environmental modifications that would allow participants to make safe choices. PMID- 16251012 TI - Validity of a 12-item version of the CES-D used in the National Longitudinal Study of Children and Youth. AB - This validation study assessed the degree of confidence that can be placed on inferences from depressive symptoms among adolescents, based on a 12-item version of the Centre for Epidemiological Studies Depression scale (CES-D). This short version of the scale had been developed for application in the National Longitudinal Study of Children and Youth and we refer to it as the CES-D-12 NLSCY. The major data source for the present validation study was a 2002/2003 survey of 12,990 students in junior and senior high school in the Atlantic provinces of Canada. Receiver operating characteristic curve analyses for two different proxy gold standards yielded adequate areas under the curve (AUCs) of .84 and .80, allowing us to establish cut points for three categories of depressive symptoms in the general adolescent population: Minimal (CES-D-12-NLSCY total score 0 to 11), Somewhat Elevated (total score 12 to 20) and Very Elevated (total score 21 to 36). The CES-D-12-NLSCY was found to have acceptable internal consistency (Cronbach=s alpha .85). All but one of the 12 items of the CES-D-12 NLSCY were found to have acceptable discrimination ability. The prevalence of Minimal, Somewhat and Very Elevated depressive symptoms in the adolescent student population of the Atlantic provinces was estimated to be 72.3, 19.5 and 5.5 percent, respectively. A further 2.6 percent of students who responded to fewer than 11 items of the scale were classified as Indeterminate with regards to depressive symptom category. The major threat to the accuracy of the CES-D-12 NLSCY is its lack of inquiry about irritability, which is a key symptom of depression in youth. PMID- 16251013 TI - An analysis of the effect of selection bias on the association of hormone replacement therapy and breast cancer risk. AB - A sensitivity analysis was conducted to determine the impact on measures of effect of a suspected differential participation response rate between hormone replacement therapy (HRT) users and nonusers, among controls recruited to a population-based case-control study of breast cancer. The age-specific prevalence of current HRT use among controls was compared to data from the 1996 Canadian National Population Health Survey (NPHS). Control women identified as current HRT users were randomly re-sampled to replicate the prevalence of HRT use reported by the NPHS. Unconditional logistic regression was conducted to estimate odds ratios (OR) and 95 percent confidence intervals (CI) for the use of HRT and breast cancer risk before and after re-sampling. Multivariate adjusted ORs for breast cancer and estrogen-only and estrogen-progestin formulations were 0.76 (0.53 1.10) and 0.94 (95% CI: 0.64 - 1.38), respectively, using the original case control controls and 0.99 (0.77-1.27) and 1.57 (95% CI: 1.02 - 2.40), respectively, following re-sampling of the controls. This sensitivity analysis illustrates the extent to which differential participation rates between HRT users and nonusers may affect estimates of measures of effect. PMID- 16251014 TI - A comparison of measures of socioeconomic status for adolescents in a Canadian national health survey. AB - The purpose of this study was to explore and compare measures of socioeconomic status (SES) in a national sample of Canadian adolescents. Issues of missing data and interrelationships among the measures were addressed. Measures of SES included household income, parental education, two parental occupation-based measures, and four neighbourhood proxy indicators. The proportion of adolescents with missing data was largest for household income (21.1 percent). Data were not missing at random, as adolescents missing household income information were less likely to reside in a high income neighbourhood. Pair-wise Spearman correlations ranged from: 0.40-0.79 between neighbourhood SES measures; 0.12-0.37 between household/parental and neighbourhood indicators; and 0.36-0.87 between household/parental measures. Correlations were lower among rural adolescents, particularly for the neighbourhood SES measures. The results highlight both measurement and conceptual challenges for researchers who wish to gain insight into SES-health relationships for adolescents. In particular, the findings emphasize the importance of incorporating multiple measures of SES and suggest a need to further explore the meaning of socioeconomic position for this population. PMID- 16251015 TI - [Establishment of a rhesus haploidentical hematopoietic stem cell and mesenchymal stem cell transplantation model by nonmyeloablative conditioning]. AB - OBJECTIVE: To establish rhesus haploidentical hematopoietic stem cell transplantation model by nonmyeloablative conditioning, and examine the effects of mesenchymal stem cells (MSC) in haploidentical transplantation. METHODS: The recipient haploidentical rhesus monkeys were conditioned with a nonmyeloablative regimen consisted of fludarabine, cyclophosphamide, 200 cGy total body irradiation, and rabbit anti-human thymocyte globulin. Cyclosporine A, mycophenolate mofetil and anti CD25 antibody were used for graft versus host disease (GVHD) prophylaxis. Rhesus monkeys in one group were given hematopoietic stem cell (HSC) only, while in the other group HSC combined with MSC. The differences in hematopoiesis recovery, chimerism level, and GVHD between the two groups were evaluated. RESULTS: Stable chimerism could be achieved in recipient monkeys. Hematopoiesis recovery was mainly related with chimerism level. MSC seemed capable of facilitating HSC engraftment, as there were more mixed chimerism and less GVHD occurrence in the HSC combined with MSC recipient group. CONCLUSION: A rhesus haploidentical hematopoietic stem cell transplantation model is successfully established by nonmyeloablative conditioning. MSC was of great benefit to haploidentical transplantation. PMID- 16251016 TI - [HLA-identical sibling allogeneic hematopoietic stem cell transplantation for chronic myelogenous leukemia in first chronic phase. Analysis of 51 cases]. AB - OBJECTIVE: To evaluate the treatment outcome of HLA-identical sibling allogeneic hematopoietic stem cell transplantation (allo-HSCT) for chronic myelogenous leukemia (CML) patients in first chronic phase (CP(1)). METHODS: Fifty-one patients with CML-CP(1) received HLA-identical sibling allo-HSCT with conditioning regimens of TBI plus Cy or Bu plus Cy. Allogeneic peripheral blood stem cell transplantation (PBSCT) and bone marrow transplantation (BMT) were performed for 28 and 23 patients, respectively. The median follow-up duration was 1434 (60 - 4062) days. RESULTS: Fifty (98.0%) patients were successfully engrafted. Transplant-related mortality occurred in 8 (15.7%) patients. Acute graft-versus-host disease (aGVHD) occurred in 35 (68.6%) patients and 11 (21.6%) patients were grade II-IV, while chronic GVHD (cGVHD) did in 17 (37.8%) patients. Five (7.4%) patients relapsed. The 5-year probability of disease-free survival (DFS) was (79.2 +/- 6.4)%. There was no significant difference in 5-year DFS, death rate and treatment related syndromes between the two conditioning regimens (P > 0.05), and in 5-year DFS, relapse rate and death rate between two transplant choices (P > 0.05). However, the rate of relapse was lower in Bu/Cy group (P < 0.01) and the rate of cGVHD was higher in allo-PBSCT group (P < 0.05). CONCLUSIONS: Allo-HSCT can cure a significant proportion of patients with CML CP(1). There was no significant difference in DFS between the two different conditioning regimens and between the different transplant choices. Donor lymphocyte infusion is a therapeutic alternative for CML patients relapsed after transplantation. PMID- 16251017 TI - [The distribution of donor hematopoietic stem cell and the ratio of lymphocytes from donor origin to recipient origin in recipient mice after allogeneic bone marrow transplantation]. AB - OBJECTIVE: To explore the hematopoietic stem cell distribution and lymphocyte proliferation and differentiation in recipient mice after allogeneic bone marrow transplantation (allo-BMT). METHODS: BALB/c (H-2(d)) mice were total body irradiated 5.5 Gy x 2 by (137)Cs and then transplanted with bone marrow cells from GFP transgenic C57BL/6J (H-2(b)) mice. The femur, spleen, Peyer patches, thymus, liver and peripheral blood of the host were collected on days 3, 7, 21, 35 and 70 post transplantation, and their sections were observed by fluorescent microscopy. The green fluorescent cells were counted with FACS. The phycoerythrin (PE) labeled antibodies to CD4, CD8 and B220 were used for sorting T and B lymphocytes. RESULTS: (1) On day 3 and day 7 after allo-BMT, there were (1.06 +/- 0.02)% and (76.60 +/- 1.80)% of donor's green bone marrow cells in host's spleen respectively, whereas only (0.37 +/- 0.06)% and (39.70 +/- 5.38)% in the bone marrow, respectively. (2) In bone marrow and other organs of 21 day-old chimerism mice, over 60% cells were of donor origin. (3) There were (0.36 +/- 0.04)% donor's bone marrow cells lodging at host's Peyer patches, similar to that in bone marrow. CONCLUSION: (1) The engrafted allogeneic hematopoietic stem cell can move into spleen, bone marrow, Peyer patches and thymus. The spleen is the main lodging place of the engrafted cells early after all-BMT. (2) The majority of cells in chimerism mice immunologic organs were of donor origin. (3) Peyer patches is another lodging place early after allo-BMT. PMID- 16251018 TI - [Effects of mesenchymal stem cells on expansion potential and adhesion molecules expression of cord blood CD34+ cells]. AB - OBJECTIVE: To explore the effects of bone marrow mesenchymal stem cells (MSCs) on in vitro expansion potential, the adherent molecules expression of cord blood (CB) CD34(+) cells. METHODS: MSCs were obtained from human bone marrow and their differentiation function and phenotype were identified. CB CD34(+) cells were expanded in culture systems with or without MSC layer. Hematopoietic progenitor cells and adhesion molecules expression were assessed by semisolid culture assay and flow cytometry. RESULTS: Thy-1, SH2, SB10, CD44, CD13, CD49e and CD29 were highly expressed on MSCs with no expressions of CD34, CD45, HLA-DR, CD14 and CD31. The MSCs could differentiate into adipocytes and osteoblasts under specific induction conditions. After culturing on MSCs layer with supplement of cytokines for 8 days, the absolute numbers of nuclear cells, CD34(+), CD34(+)CD38(-), CD34(+)CD62L(+) cells and CFU-Cs were increased by 145.57 +/- 17.89, 37.47 +/- 13.78, 69.78 +/- 50.07, 10.74 +/- 5.89 and 20.73 +/- 5.54-folds, respectively, being significantly higher than that cultured with cytokines alone. The expression of ALCAM, VLA-alpha4, VLA-alpha5, VLA-beta1, HCAM, PECAM and LFA-1 on CD34(+) cells remained unaffected. The expressions of ICAM-1 and L-selectin were downregulated during expansion, while the absolute numbers of CD34(+)CD62L(+) and CD34(+)CD54(+) cells were increased. CONCLUSIONS: MSCs layer improves expansion of CB CD34(+) cells while inhibiting their differentiation and retaining their homing ability. PMID- 16251021 TI - [The effect of HGF on graft-versus-host disease and graft-versus-leukemia after allogeneic bone marrow transplantation in acute lymphoblastic leukemia mice]. AB - OBJECTIVE: To investigate the effect of hepatocyte growth factor (HGF) on graft versus-host disease (GVHD) and graft-versus-leukemia (GVL) after allogeneic bone marrow transplantation (allo-BMT) and related mechanism in acute lymphoblastic leukemia (ALL) mice. METHODS: Twenty nude mice were randomly divided into control (group A) and test (group B) groups for monitoring relapse, and 20 BALB/c mice into control (group C) and test (group D) groups for GVHD. HGF as injected from day 0 to day 7 after BMT for groups B and D, while PBS for A and C. CD4(+) and CD8(+) T cell were evaluated by flow cytometry. The survival of mice after BMT was recorded. The level of tumor necrosis factor-alpha (TNF-alpha) was evaluated by ELISA. RESULTS: The median past-BMT survival were 7.00 +/- 1.58, 9.00 +/- 1.58, 11.00 +/- 3.95 and 24.00 +/- 13.44 days for groups A, B, C, D, respectively, being prolonged in group D. HGF could decrease the quantity of CD4(+) T cells [group D (10.39 +/- 1.15)% vs group C (13.50 +/- 1.80)%, P < 0.01] and increase CD8(+) T cell [group D (12.25 +/- 2.85)% vs group C (6.12 +/- 1.99)%, P < 0.01], decrease the level of TNF-alpha in transplanted ALL mice [group D (112.10 +/- 18.99) pg/ml vs group C (143.90 +/- 25.35) pg/ml, P < 0.01] and reduce the degree of GVHD. CONCLUSION: HGF could alleviate post-allo-BMT GVHD but retain GVL effect. PMID- 16251023 TI - [Study on rapid generation of dendritic cells from K562 cell line induced by A23187 alone]. AB - OBJECTIVE: To explore a simple, rapid and efficient way to generate dendritic cells from leukemic cells. METHODS: K562 cells were cultured with calcium ionosphere A23187 alone, A23187 plus GM-CSF, or a DC differentiation cocktail consisting of GM-CSF, IL-4 and TNF-alpha, respectively. The expression of surface markers of induced DCs was analyzed by flow cytometry. The K562-DCs stimulating the proliferation of allo-genetic naive T cells and inducing cytotoxicity of T cells were determined by MTT assay. RESULTS: Microscopic examination revealed that under all the three culture conditions, K562 cells became displaying DC morphology. At 72 hours in the two culture systems containing A23187, there were higher proportions of cells with dendritic morphology [(69.5 +/- 17.2)% and (73.1 +/- 13.9)%, respectively] than that in the cocktail system [(28.5 +/- 12.3)%] (P < 0.05). And the same did when cultured for 7 days [(69.5 +/- 17.2)%, (73.1 +/- 13.9)% respectively vs (51.2 +/- 10.7)%, P < 0.05]. In the 7-day cultures, the percentage of CD1a expressing cells was lower [(8.2 +/- 2.3)% and (10.3 +/- 5.1)% vs (17.2 +/- 1.6)%, respectively] while the CD83 expressing cells was higher [(85.6 +/- 8.8)% and (82.4 +/- 9.1)% vs (77.4 +/- 12.9)%, respectively] compared with that in the cocktail system (P < 0.05). No significant difference was found in the allogeneic T cell proliferation response and induced T cell cytotoxicity between A23187 containing and cocktail groups (P > 0.05). CONCLUSIONS: A23187 treatment is a simple, rapid and efficient in vitro strategy for inducing dendritic cell from leukemic cells. PMID- 16251019 TI - [Successful treatment of agammaglobulinemia by HLA-mismatched unrelated cord blood stem cell transplantation--the first case report]. AB - OBJECTIVE: To evaluate cord blood stem cell transplantation (CBT) in the treatment of X-linked agammaglobulinemia, and observe the courses of the hematopoietic and immune reconstitution. METHODS: A 14-year-old male patient with agammaglobulinemia received CBT from a 1/6 HLA-mismatched unrelated cord blood. The conditioning regimen was Bu/Cy/anti-CD3 antibody. CsA was given together with MMF and MTX for prophylaxis of GVHD. The patient received 0.42 x 10(8) nucleated cells/kg, containing 0.35 x 10(6) CD34(+) cells/kg. RESULTS: The recipient showed hematopoietic reconstitution on day 30 post-transplantation when ANC was 0.5 x 10(9)/L and BPC 20 x 10(9)/L. Sex chromosome analysis showed engraftment (donor 46, XX/recipient 46, XY = 4:1) on day 45. The recipient's blood group changed from AB to O, IgG from 1.1 g/L to 3.5 g/L, sex chromosome from 46, XY to full 46, XX, and mature B cells in peripheral blood from 0 to 5% on day 100, indicating immune reconstitution. At the last follow-up of 360 days, the patient without acute or chronic GVHD showed normal hemogram and myelogram, IgG 13.5 g/L and 10% mature B cells in peripheral blood, indicating the hematopoiesis and immune persistent reconstitution. No acute or chronic GVHD was developed. CONCLUSION: This is the first case report of successful treatment of X-linked agammaglobulinemia by HLA-mismatched unrelated CBT. PMID- 16251024 TI - [Peripheral blood naive T cell level and its T cell receptor Vbeta repertoire usage profile in patients with chronic myelogenous leukemia]. AB - OBJECTIVE: To analyze peripheral blood naive T cell level, its T cell receptor (TCR) Vbeta repertoire usage profile and clonality for evaluating the recent thymic output function and the expansion feature of TCR Vbeta subfamily T cells in patients with chronic myelogenous leukemia (CML). METHODS: Quantitative detection of T-cell receptor excision DNA circles (TRECs) in peripheral blood mononuclear cells (PBMNC) from 20 cases of CML was preformed by real-time PCR (TaqMan) analysis, and TRECs-number in T-cells was calculated from peripheral blood CD3-positive cell rate. The expression and clonality analysis were detected by RT-PCR and Genescan technique in PBMNC from 14 out of the 20 patients. Nine normal individuals served as controls. RESULTS: A dramatic reduction of TRECs value in patients with CML was detected as compared with that in normal controls. The mean value of TRECs was 0.06 +/- 0.16 copy/1000 CD3(+) cells in CML patients while 6.84 +/- 4.71 copies/1000 CD3(+) cells in normal controls (P < 0.01). The 1 - 12 Vbeta subfamilies were variably expressed in samples from 14 patients. Genescan analysis identified clonal expanded T cells of some Vbeta subfamily from 13 cases. Vbeta3, Vbeta10, Vbeta19, Vbeta21 and Vbeta22 subfamilies clonal T cells were more frequently seen. CONCLUSION: There is a prominent reduction of recent thymic output naive T cells function in CML. The predominant usage and clonal expansion of TCR Vbeta subfamily T cells could be identified, indicating that CML patients have specific immune response to leukemia associated antigen, in spite of their T cell immunodeficiency. PMID- 16251025 TI - [Detection of RbAp46 expression in bone marrow cells of leukemia patients by real time quantitative RT-PCR]. AB - OBJECTIVE: To investigate retinoblastoma (Rb) associated protein 46 (RbAp46) gene expression levels in bone marrow (BM) cells of leukemia patients. METHODS: Real time quantitative reverse polymerase chain reaction (QRT-PCR) method was used for detecting RbAp46 expression levels in BM cells of 140 patients with acute leukemia (AL), 13 with chronic myelogenous leukemia in chronic phase (CML-CP), 7 with CML in blast crisis (CML-BC) and 32 with non-leukemic disorders. RESULTS: The M-Estimators of RbAp46 were higher in 98 newly diagnosed ALs and 5 relapsed ALs than in 28 ALs in complete remission (CR) and 32 non-leukemic controls (178.23 and 213.65 vs 85.89 and 88.08, respectively). No statistic difference was found between the CR group and control group, or between the newly diagnosed group and relapsed group. The M-Estimators of RbAp46 in patients with CML-CP was 58.27, similar to that in control, but much lower than that in CML-BC (173.24). Among 98 newly diagnosed ALs, the M-Estimators of RbAp46 in M(3) and M(4) were the lowest in all of the subtypes. Furthermore, the RbAp46 expression levels were not correlated with the expression of the fusion genes of bcr/abl, PML-RARalpha, and multidrug resistant gene (mdr1), but were positively correlated with Wilms' tumor gene (WT1) expression levels and negatively with AML1/ETO fusion gene expression. CONCLUSION: RbAp46 expression levels in ALs and CML-BC were strikingly higher than that in non-leukemias and CML-CP, and might participate in leukemogenesis. PMID- 16251026 TI - [Effects of RNA interference inhibiting SDF-1 expression in bone marrow stromal cells on the proliferation and apoptosis of co-cultured Jurkat cells]. AB - OBJECTIVE: To study the effects of RNA interference inhibiting stromal cell derived factor-1 (SDF-1) expression on the proliferation and apoptosis of co cultured Jurkat cells. METHOD: Inhibition of SDF-1 expression by RNA interference (RNAi) was achieved by transferring SDF-1 specific short hairpin RNA (shRNA) expressing plasmid into cultured human acute leukemic bone marrow stromal cells. Resistant clones were obtained by G418 selection (group A). The concentration of SDF-1 protein in culture supernatant was detected by enzyme-linked immunosorbent assay (ELISA). The population double time (PDT), cell cycles, apoptosis rates and the expressions of PCNA, Bcl-2/Bax, Fas/FasL of co-cultured Jurkat cells were detected by cells counting, flow cytometry. TdT-mediated dUTP nick-end labelling (TUNEL) and immunocytochemistry (ICC), respectively. The un-transfected acute leukemic (group B) and normal (group C) bone marrow stromal cells were taken as controls. RESULTS: The content of SDF-1 protein in supernatant of group A?[(384 +/- 41) pg/ml] was significantly lower than that in group B[(2474 +/- 271) pg/ml] or group C[(1324 +/- 154) pg/ml]. As group A compared with group B and group C, the PDT of co-cultured Jurkat cells was prolonged (group A: 42 h, vs group B: 29 h, group C: 33 h), and G(0)/G(1) stage cells increased [group A: (28.47 +/- 2.39)%, vs group B: (19.43 +/- 2.80)%, group C: (27.15 +/- 2.07)%], S stage cells decreased [group A: (25.57 +/- 1.90)%, vs group B: (74.48 +/- 3.23)%, group C: (60.99 +/- 2.33)%], G(2)/M stage cells increased [group A: (45.96 +/- 3.24)%, vs group B: (6.09 +/- 1.96)%, group C: (11.86 +/- 1.98)%], the apoptosis rate increased [group A: (15.2 +/- 0.8)%, vs group B: (5.4 +/- 0.7)%, group C: (9.5 +/ 0.4)%], and the expressions of PCNA, Bcl-2, Fas decreased; whereas the expressions of Bax and FasL were increased. CONCLUSION: The inhibition of SDF-1 expression in bone marrow stromal cells inhibits the proliferation and promotes the apoptosis of co-cultured Jurkat cells. PMID- 16251038 TI - [Collagen type III glomerulopathy: a morphologic study]. AB - OBJECTIVE: To study the morphologic changes of collagen type III glomerulopathy and to investigate the possible cellular origin for collagen III production. METHODS: Light microscopy, immunofluorescent staining, immunohistochemistry (for collagen I, III and IV and alpha-SMA) and electron microscopy studies on 3 renal biopsy cases of collagen type III glomerulopathy were performed. RESULTS: Two cases presented with nephrotic syndrome, one of which was associated with systemic hypertension. The third case showed renal impairment and renal hypertension. None had any known family history of renal diseases. Light microscopy showed diffuse thickened glomerular basement membrane and expanded mesangium with deposition of weakly PAS-positive homogeneous material not associated with mesangial cell proliferation. Electron microscopy revealed massive collagen fiber deposits in the subendothelial spaces and mesangium. The mesangial cells also contained bundles of microfilaments in the subplasmalemmal regions. Immunohistochemically, the diffuse positivity for type III collagen corresponded to the homogeneous material seen under light microscopy. The staining for type I and IV collagens was negative. Alpha-SMA was expressed in many mesangial cells. CONCLUSIONS: The diagnosis of collagen type III glomerulopathy can be made on the basis of detailed morphologic examination and ancillary investigations. It is possible that activated mesangial cells may be the cellular origin of collagen III. PMID- 16251039 TI - [Mucinous noncystic (colloid) adenocarcinoma of the pancreas]. AB - OBJECTIVE: To determine the clinicopathologic characteristics and the relationship between related gene expression and pathobiologic behavior of pancreatic mucinous noncystic adenocarcinoma. METHODS: Among the 249 pancreatic carcinoma cases from the department files, 6 tumors were identified to meet the pathologic criteria of colloid carcinoma. Envision immunohistochemical staining technique was used to detect expression of p21(ras), p21(WAF1), p16, p33(ING1), p53, ATM, MDM2, PCNA, Cyclins (D1, D3, A, B and E). Intra- and extra- cellular mucin production were determined by AB-PAS staining. Clinically, all of 6 cases were followed to June, 2003. RESULTS: In all 6 cases, the tumors were located in the head of the pancreas and all displayed similar microscopic findings. Duodenal invasion was seen in 4 cases and perineural invasion was seen in 1 case. Tumor metastasis in the liver was seen in 2 cases and in the regional lymph nodes in 2 cases. Positive immunostaining was seen in 5 cases with p21(ras), 3 cases with p21(WAF1), 1 case with p16, 4 cases with p33(ING1), 2 cases with p53, 3 cases with ATM, 3 cases with MDM2, 6 cases with PCNA, 3 cases with cyclinA, 3 cases with cyclinD1, 4 cases with cyclinD3, 4 cases with cyclinB and 6 cases with cyclinE. Both extracellular and intracellular mucin was strongly positive for AB PAS staining. Clinical follow-up found that 2 patients died of their tumors at 14 and 20 months. Three patients were alive after 28, 49 and 87 months of follow-up. One case were lost contact. CONCLUSIONS: Pancreatic mucinous noncystic adenocarcinoma has distinct morphologic features and biologic behavior. Multiple gene products including many cyclins may be involved in the pathogenesis of pancreatic colloid carcinoma. The tumor has an aggressive behavior with a high frequency of invasion and metastases, though the prognosis could be better than that of ordinary ductal adenocarcinoma of pancreas. PMID- 16251040 TI - [Hemangioblastoma of the optic nerve: a case report]. PMID- 16251041 TI - [Expression of macrophage inflammatory protein-1alpha, a disintegrin-like and metalloproteinase 8 and 12, and CD68 protein in giant cell lesions of jaw and giant cell tumors of long bone]. AB - OBJECTIVE: To detect the expression of macrophage inflammatory protein-1alpha (MIP-1alpha), a disintegrin-like and metalloproteinase (ADAM) 8 and 12 and CD68 protein in giant cell lesions of jaw and giant cell tumors of long bone, and to study their effects on the histogenesis of giant cells in such lesions. METHODS: MIP-1alpha, ADAM8, ADAM12 and CD68 were detected by immunohistochemistry in 24 paraffin-embedded specimens of central giant cell lesions of jaw and giant cell tumors respectively. RESULTS: MIP-1alpha positive signal was located in blood vessels and bone. ADAM8, ADAM12 and CD68 positive signals were located in the cell membrane and cytoplasm of all multinucleated giant cells and some round mononuclear cells in the lesions. In addition, some spindle mononuclear stromal cells were positive for ADAM12 in both lesions. CONCLUSION: Multinucleated giant cells probably originate from CD68-postive round mononuclear cells, which are recruited from monocyte-macrophage system by chemokines, such as MIP-1alpha, followed by cell fusion mediated by ADAM8 and ADAM12. PMID- 16251042 TI - [Ectopic hamartomatous thymoma: a clinicopathological and immunohistochemical study of two cases]. AB - OBJECTIVE: To study the clinicopathological and immunohistochemical features of ectopic hamartomatous thymoma (EHT), and to discuss its histogenesis. METHODS: The clinical and pathologic features of two EHT cases of were evaluated. Immunohistochemical study was performed by LSAB method using a panel of antibodies including AE1/AE3, CK5, CK7, CK8, CK20, EMA, vimentin, CD5, CD10, alpha-SMA, calponin, desmin, CD34, S-100 protein, CD57, GFAP, TTF-1 and CD99. RESULTS: Both cases occurred in males aged 20 years and 40 years respectively. Each patient presented with a solitary mass, one located in the suprasternal fossa and the other in the left supraclavicular region for a period of 6 months and 2 months respectively. Grossly, the masses were well-circumscribed with spherical and ovoid appearance, measuring 5 cm and 3 cm in maximum diameter respectively. On cut section, they were gray-white in color and of soft consistency. Histologically, both tumors were composed of a mixture of spindle cells, epithelial cells and mature adipose tissue. The spindle cells element accounted 85% and 70% each in the two cases. They resembled fibroblasts in morphology and were arranged frequently in fascicular, woven or storiform patterns. Epithelial cells element represented nearly 10% in both cases. Most of the epithelial cells had a non-keratinization squamous appearance. They formed small solid islands and adamantinoma-like "nastomosing cords", or appeared as lining cells in large cystic spaces. In focal areas, glandular differentiation presented as small glands. A transition between the spindle cell and epithelium components could be also identified in some areas. Mature adipose tissue was irregularly distributed in the two tumors, about < 5% and 20% respectively. Immunohistochemically, the epithelial element expressed AE1/AE3, CK5, CK7, CK8 and EMA, whereas the spindle component expressed AE1/AE3, CK5, CK7, CK8, vimentin, CD10, CD34, alpha-SMA, MSA, and calponin. Both elements were negative for CK20, TTF-1, desmin, S-100 protein, CD57, GFAP and CD99. CONCLUSIONS: EHT is a benign tumor that occurs predominantly in the lower neck region of young to middle-aged males. Immunohistochemical study revealed myoepithelial differentiation of the spindle cells, suggesting EHT is a mixed tumor composed of epithelial and myoepithelial cells. EHT possibly originates from the remnants of cervical sinus of His, and therefore, may be renamed as branchial anlage mixed tumor. PMID- 16251043 TI - [Localized thymic Langerhans cell histiocytosis with myasthenia gravis]. PMID- 16251044 TI - [Inhibitory effects of transfected Bcl-XL antisense oligodeoxynucleotide on proliferation of esophageal cancer cells and growth of human esophageal carcinoma in nude mice]. AB - OBJECTIVE: To investigate the biologic effects of Bcl-XL antisense oligodeoxynucleotide (ASODN) transfected into cultured esophageal carcinoma cells and human esophageal carcinoma xenograft in nude mice. METHODS: Cationic liposome mediated ASODN was used to transfect esophageal carcinoma cells. RT-PCR, Western blot, MTT assay, flow cytometry and in-situ apoptosis cells detection (TUNEL detection) were used to systematically study the biological effects of the transfected cells in vitro and in vivo. RESULTS: MTT assay showed that the proliferation of esophageal carcinoma cells in the ASODN group decreased significantly as compared with control (P < 0.05), along with a 57.3% inhibitory rate of Bcl-XL mRNA, a significant decrease of Bcl-XL protein and the apoptosis rates of (31.1 +/- 5.8)% and 35.0% by flow cytometry and TUNEL assay, respectively (P < 0.01, as compared with controls). The growth of human esophageal carcinoma in nude mice was also significantly inhibited in the ASODN group (P < 0.05), along with a significant decrease of Bcl-XL mRNA and protein expression, and also an enhanced apoptosis of the tumor cells in nude mice. CONCLUSIONS: Bcl-XL ASODN can effectively inhibit the proliferation of esophageal carcinoma cells in vitro and the growth of the tumor in vivo. The suppression of Bcl-XL expression by ASODN may offer both a therapeutic approach and an important theoretic foundation for gene therapy against esophageal carcinoma. PMID- 16251045 TI - [Effects of transforming growth factor-beta/Smad signaling on the growth and apoptosis of human rhabdomyosarcoma cell line RD]. AB - OBJECTIVE: To study the effects of TGF-beta/Smad signaling on the growth and apoptosis of human rhabdomyosarcoma cell line RD. METHODS: Biosynthesized short hairpin RNA (shRNA) was transfected into RD cells by cation liposome vector to suppress Smad4 expression. The mRNA and protein expression of Smad4 in RD after shRNA-transfection were examined by RT-PCR and Western blot respectively. Immunofluorescent staining was used to detect the location of Smad2/3 in RD by laser scanning confocal microscopy. The viability of RD cells was examined by MTT method and (3)H-thymidine incorporation assay. The apoptosis of RD was examined by flow cytometry analysis and fluorescent staining. RESULTS: The expression of mRNA and protein of Smad4 in RD were effectively suppressed by shRNA interference. Such suppression effectively interrupted the endogenous TGF beta/Smad signaling and consequently blocked the translocation of Smad2/3. The interruption of endogenous TGF-beta/Smad signaling not only inhibited the growth of RD but also induced apoptosis of RD. Exogenous TGF-beta1 inhibited the growth of RD but did not influence the apoptosis of RD. CONCLUSION: shRNA interference can effectively interrupt the TGF-beta/Smad signaling by suppressing the expression of Smad4. TGF-beta/Smad signaling subtly regulates the growth and apoptosis of RD. PMID- 16251046 TI - [Influence of Smad4-independent pathway of transforming growth factor beta1 on the biological activity of pancreatic cancer cells]. AB - OBJECTIVE: To study effects of the expression of transforming growth factor (TGF) beta1 on the growth of Smad4-null pancreatic cancer cells. METHODS: TGF-beta1 eukaryotic expression vector was transfected into pancreatic cancer cell line BxPC3. Effects of the expressison of TGF-beta1 was studied by growth curve analysis and flow cytometry. Cell motility was monitored by wound-healing assay. Western blot was used to estimate the expression level of p21(WAF/CLIP1), a cyclin-dependent kinase inhibitor. RESULTS: Transfection of TGF-beta1 changed the morphology of BxPC3 into spindle shaped cells. The growth rate of BxPC3 began to decrease after the fourth day of TGF-beta1 transfection, compared with the control groups. Flow cytometry showed that the percentages of cells in the S phase were (27.53 +/- 0.02)%, (26.32 +/- 0.01)% and (17.01 +/- 0.03)% in naive BxPC3, vector-control group and TGF-beta1 transfection group respectively. Lesser cells entered the S phase after TGF-beta1 transfection (P < 0.01), but no difference was seen between the BxPC3 and vector groups (P > 0.05). The expression of p21(WAF/CLIP1) increased upon the expression of TGF-beta1. CONCLUSION: The Smad4-independent pathway of TGF-beta1 not only induces epithelial-mesenchymal transition in pancreatic cancer BxPC3, but also inhibits its growth through the up-regulation of p21(WAF/CLIP1). PMID- 16251047 TI - [Effects of aldose reductase transfection on the proliferation of rat mesangial cells in vitro]. AB - OBJECTIVE: To study the effects of aldose reductase (AR) on the proliferation of rat mesangial cells (MsC) in vitro and to investigate its mechanism. METHODS: Cell proliferation was assessed by MTT colorimetric assay. Cell cycle and apoptosis were analyzed by flow cytometry. The growth of normal MsC and AR transfected MsC was compared. The proliferation of PDGF-BB and cellular growth stimulation by 10% NBS were investigated using AR inhibitors (ARI) Sorbinil and Zopolrestat. The effects of PDGF-BB on the expression of AR, p65 and c-Jun were assessed by Western blot. Activation of AP-1 was measured by EMSA. RESULTS: AR expression of transfected MsC was distinctly higher than that of the control. Transfected MsC grew quicker than normal cells. ARI partially inhibited the proliferation of transfected MsC under the stimulation of PDGF-BB and 10% NBS, whereas 10% NBS had no effect on normal MsC. PDGF-BB upregulated the expression of AR and c-Jun, but had no effect on p65. The upregulation of c-Jun and the activation of AP-1 could be attenuated by ARI. CONCLUSION: AR may participate in the pathological proliferation of MsC through the pathway related to the activation of AP-1. PMID- 16251048 TI - [Humoral immune response and in vitro neutralizing antibody assay on co-delivery of protein HPV16L1 virus-like particle with HPV16L1 gene]. AB - OBJECTIVE: To compare humoral immune response by co-inoculating mice with antigen HPV16L1 virus-like particle (VLP) and HPV16L1 recombinant plasmids and then observing the neutralizing antibody activity in vitro. METHODS: C57BL/6 mice were injected intramuscularly/subcutaneously with pcDNA-L1 plasmids plus HPV16L1 VLP. Serum IgG levels were detected by ELISA, antibody neutralizing protective activities were determined by hemagglutination inhibition and HPV16L1 VLP binding inhibition assay. RESULTS: Serum antibody titers and neutralizing antibody activities were increased in HPV16L1 plasmids plus HPV16L1 VLP proteins in co immunized mice when compared with controls. CONCLUSION: Co-inoculation of the HPV16L1 VLP protein can enhance production of neutralizing antibody activities against aimed antigen, which should be a more promising strategy for effective HPV16 prophylactic vaccine development. PMID- 16251049 TI - [Large mediastinal angiomyolipoma: a case report]. PMID- 16251050 TI - [Homocysteine promotes endothelial cells to express macrophage inflammatory protein-1alpha]. PMID- 16251051 TI - [Human nasal type NK/T cell lymphoma: study of differential expression genes by cDNA microarray]. PMID- 16251052 TI - [Pathologic observation of spleen dendritic cells in mouse multiple organ failure model]. PMID- 16251053 TI - [Attractin]. PMID- 16251054 TI - [Tissue-specificity of the endothelial system]. PMID- 16251055 TI - [Diagnostic and prognostic use of DNA-Image-Cytometry in cervical squamous intraepithelial lesions and invasive carcinoma]. PMID- 16251056 TI - [Comparison of the liquid-based preparation in sputum specimens and the conventional smears]. PMID- 16251057 TI - [Nodular lymphocyte predominant Hodgkin lymphoma]. PMID- 16251058 TI - [Ovarian endometrial adneocarcinoma with choriocarcinomatous differentiation: a case report]. PMID- 16251059 TI - [Lipoprotein glomerulopathy: a case report]. PMID- 16251060 TI - [Low-grade (fibromatosis-like) spindle cell carcinoma of the breast: case report and review of the literature]. PMID- 16251062 TI - [Prevention of complications in liver transplantation]. PMID- 16251063 TI - [Perioperative management in liver transplantation patients]. PMID- 16251064 TI - [What are the indications of liver transplantion in hepatic cancer patients?]. PMID- 16251066 TI - [Biliary complications following early hepatic arterial insufficiency in liver transplantation]. AB - OBJECTIVE: To explore the clinical feature and treatment efficiency of patients with early hepatic arterial insufficiency (HAI) and biliary complications (BC) following liver transplantation (LT). METHODS: The clinical data of 240 patients receiving LT from February 1999 to February 2004 were analyzed retrospectively. End-to-end choledococholocostomy was applied to 236 patients as the major biliary reconstructive method. Hemodynamic monitoring of the hepatic artery was performed to discover HAT, including hepatic arterial thrombosis (HAT) or hepatic arterial stenosis (HAS) in the first 3 months after transplantation. RESULTS: In HAI Group, 7 cases of stricture of biliary tract and 4 cases of biliary leakage occurred; and 6 cases underwent endoscopic and/or intervention treatment, 4 cases underwent repair of anastomotic stoma and biliary drainage, 1 case underwent medication. Eight patients died and 3 were cured. A total of 32 patients (13.3%) developed biliary complications (BCs), Eleven of the 32 patients with BCs, biliary stricture in 8 cases and biliary leakage in 3 cases, had early HAI (HAI Group) with an incidence of 4.6% for BC with background of HAI (11/236). Another 21 patients with BCs (21/236, 8.7%) did not showed background of HAI (non-HAI Group). Preoperative serum total bilirubin levels were 373.3 +/- 93.9 micromol/L in HAI Group and 110.8 +/- 45.0 micromol/L in non-HAI Group (P = 0.008). Three cases with HAT underwent emergency thrombectomy then the hepatic arterial flow turned to normal. Two cases with HAS received short-term anticoagulant therapy. Recipients with BC underwent radiological and/or endoscopic interventional treatment (n = 6), surgical repair of leak site and biliary drainage (n = 4), and ordinary medication (n = 1). The 1 and 3-year survival rates of HAI Group were 54.6% and 16.4% respectively; both significantly lower than those of non-HAI Group (66.3% and 61.2%, both P = 0.042). CONCLUSION: The recipients with BC following early HAI are associated with poor outcome. The monitoring protocol of hepatic hemodynamics by color dopplar ultrasonography should be enhanced, which contributes to urgent revascularization or recovery of hepatic hemodynamics. Interventional therapies should be immediately applied and transferred to surgical treatment or even retransplantation when necessary for patients with BC following early HAI. PMID- 16251067 TI - [Diagnosis and management for early hepatic artery thrombosis after liver transplantation]. AB - OBJECTIVE: To explore the diagnosis and management of early hepatic artery thrombosis (HAT) after liver transplantation. METHODS: Routine examination of Color Doppler Imagine (CDI) was used to detect hepatic artery flow after liver transplantation in 220 cases from April 2001 to November 2004. Suspected patients were further confirmed by immediate hepatic artery angiography, and continuous infusion of urokinase through hepatic artery with catheter was performed to the patients with HAT. RESULTS: HAT was identified in 6 patients (2.7%), occurring 5.5 days (2 - 19 days) after liver transplantation. Hepatic artery recanalization was obtained in 6 cases. One patient died from lung infection 2 months after liver transplantation. One patient underwent the second liver transplantation because of the recurrence of HAT 6 months after the first transplantation, but died from multiple system organ failure 2 months after the operation. The other 4 cases have been surviving well disease-freely. CONCLUSIONS: CDI is useful for the diagnosis of early HAT after liver transplantation. Contributes to recovery of the hepatic artery flow, continuous infusion of urokinase through hepatic artery with catheter may be the first choice of the therapy for early HAT. PMID- 16251069 TI - [Effect of heme oxugenase-1 on delayed xenograft rejection: experiment of guinea pig-to-rat liver xenotransplantation]. AB - OBJECTIVE: To investigate the effects and mechanism of heme oxygenase-1 (HO-1) in liver xenotransplantation and mechanism thereof. METHODS: Thirty male guinea-pigs used as donors were injected intravenously with cobra venom factor (CVF) and then randomly divided into 3 groups 24 hours after: Group A injected intraperineally with NaCl, Group B injected intraperineally with cobalt-protoporphyrin (CoPP), heme oxygenase-1 inducer, and Group C injected intraperineally with CoPP and zinc protoporphyrin (ZnPP), HO-1 inhibitor zinc before their livers were harvested. Thirty male SD rats used as recipients underwent the above-mentioned treatment 24 hours before receiving the xenografts. Five pairs of guinea pigs and rats in each group underwent collection of blood and liver tissues 3 hours after the recovery of blood perfusion in the transplanted livers for detection of serum enzymes by biochemical methods and expression of HO-1 mRNA and protein in the transplanted livers by RT-PCR and Western blotting respectively. The other 5 pairs in each group were used to observe the survival time. RESULTS: The survival time of Group B was 15.5 h +/- 3.8 h, significantly longer than those of Group A (7.3 h +/- 2.1 h) and Group C (6.7 h +/- 2.9 h, both P < 0.01). The values of ALT and LDH of Group B were significantly lower than those of Group A and C (all P < 0.05). HOI 1 mRNA expression was not detected or only expressed in trace amount in the livers of normal guinea pigs, expressed in a small amount in the transplanted livers of Group A. The expression of HO-1 mRNA and that of HO-1 protein in the transplanted livers of Group B were significantly higher than those of Group A (both P < 0.01), and the expression of HO-1 mRNA and that of HO-1 protein in the transplanted livers of Group C were not significantly different from those of Group A (both P > 0.05). Remarkable NF-kB band was detected in Groups A and C, and only weak NF-kB band was seen in Group B. The E-selectin expression was significantly lower in the transplanted livers of Group B than in those of Group A and C (both P < 0.05). CONCLUSION: HO-1 delays the occurrence of delayed xenograft rejection in liver xenotransplantation. This effect depends, at least in part, on HO-1-mediated inhibition of endothelium activation in xenografts. PMID- 16251071 TI - [The clinical study of the relationship between allergic rhinitis and allergic asthma in the patients with autumnal pollinosis]. AB - OBJECTIVE: To investigate the relationship between allergic rhinitis and allergic asthma in the patients with autumnal pollinosis. METHODS: 1120 patients with autumnal pollinosis, aged 5 - 75, excluding those with typical symptoms of seasonal rhinitis or asthma but with positive skin test and serum IgE specific to dustmite and fungi, underwent standardized clinical questionnaire survey, including the onset age, onset time, and symptoms as well as the severity of asthma, skin tests, and examination of serum IgE specific to autumnal pollens. RESULTS: The average onset age of the allergic rhinitis patients induced by autumnal pollens was 27.9 years, significantly younger than that of the allergic asthma patients (32.6 years, P < 0.001). Out of the 1120 patients 1096 (97.9%) had allergic rhinitis, 602 (53.8%) had asthma, 507 (45.3%) had allergic rhinitis only, and 10 (0.9%) had allergic asthma only. Among the 1096 patients with allergic rhinitis 585 (53.4%) suffered from seasonal asthma. Among the 602 patients with asthma 585 (97.2%) suffered from seasonal rhinitis, and 183 of the 602 patients (30.8%) needed emergency treatment. CONCLUSION: Autumnal pollens are very important causes which induce asthma during autumnal season in northern China. PMID- 16251070 TI - [The study of inhibitory peptides on T cell activation in rheumatoid arthritis]. AB - OBJECTIVE: To study the inhibitory role of altered HA308 - 317 peptides in T cell responses in patients with rheumatoid arthritis (RA). METHODS: Peripheral blood mononuclear cells (PBMC) were obtained from 27 HLA-DRB1 positive RA patients. T cell responses to altered HA308-317 peptides were examined by using (3)H incorporation assay. The level of IL-2 and IFNgamma in the supernatants was identified by an enzyme-linked immunosorbent assay. The CD69 expression on T cell surface was studied by using flow cytometry. RESULTS: Compared to wild type CII263 - 272 or HA308 - 317, altered HA308 - 317 peptides failed to stimulate T cell proliferation (P < 0.05) and production of IL-2 as well as IFNgamma and resulted in lower expression of CD69 in RA patients (P < 0.05). SI values for T cell coincubated with altered HA308 - 317 peptides and CII263 - 272 or wild type HA308 - 317 were significantly lower than coincubated with CII263 - 272 and wild type HA308 - 317 alone (P < 0.05). CONCLUSION: Substitution of TCR-binding residue of HA308 - 317 yielded weak or non-T-cell stimulating peptides, which might be potentially effective in blocking abnormal T cell responses in RA. PMID- 16251073 TI - [Application of intraoperative real-time ultrasound in the surgical treatment of cerebral arteriovenous malformation]. AB - OBJECTIVE: To investigate the value of intraoperative real-time ultrasound during surgery of cerebral arteriovenous malformations (AVMs). METHODS: Thirteen patients with cerebral AVMs, 12 supratentorial and 1 infratentorial, underwent microsurgical treatment. Intraoperative ultrasound (IOUS) with multimodality was performed before the starting of resection in order to: (1) localize and delineate the AVM, (2) differentiate the embolized and perfused parts of AVM, and (3) identify the feeding arteries and draining veins. After removal of the lesion, sonographic examination was repeated to detect the residual AVM tissue and the findings were compared with the intra- and/or post-operative angiographic finding. RESULTS: Total resection was achieved in all 13 patients with AVM under real-time ultrasonographic navigation. The niduses of AVM were localized and defined precisely by IOUS due to their dichromatic flow pattern. The embolized parts of AVM exhibited hyperechogenicity without blood signals. The feeding and draining vessels were distinguished from normal ones morphologically and hemodynamically. Complete removal of the AVM determined sonographically was then confirmed by angiography. CONCLUSIONS: Intraoperative real-time ultrasound allows accurate localization and navigation of the cerebral AVMs, provides valuable information during surgery, and helps determine the degree of removal of the cerebral AVM. PMID- 16251074 TI - [Clinical value of computer aided detection system in direct full-field digital mammography: a preliminary evaluation]. AB - OBJECTIVE: To study the clinical value of computer aided detection (CAD) in direct full-field digital mammography. METHODS: 480 digital mammograms of 120 patients with pathologically proved breast cancer were randomly selected and interpreted by an expert radiologist without knowing the pathologic diagnosis, then analyzed by CAD system, and finally read by the radiologist again to make the final diagnosis. The sensitivity for breast cancer detection of the radiologist in diagnosis of breast cancer was calculated before and after using CAD. Data of the breast density, diameters of mass and calcification, being spiculate or not, and pathology of each case were reviewed to determine how they impacted on the improvement of sensitivity with the assistance of CAD. RESULTS: The sensitivity for breast cancer detection by the radiologist before using CAD was 74.2%, significantly lower than that after using CAD (89.2%, chi(2) = 37.628, P = 0.000). Breast density (chi(2) = 7.352, P = 0.007), diameter of calcification (chi(2) = 10.549, P = 0.014) and pathologic pattern (chi(2) = 13.751, P = 0.003) were highly correlated with the improvement of sensitivity. Use of CAD might overcome type 1 missed diagnosis, and not type 2 missed diagnosis. CONCLUSION: Computer-aided detection helps radiologist improve their diagnostic accuracy of breast cancer, however, is not capable of differentiating benign from malignant lesions. PMID- 16251075 TI - [Influence of breast carcinoma cells on normal endothelial cells: experimental study with a co-culture system]. AB - OBJECTIVE: To study the influence of breast carcinoma cells on normal endothelial cells. METHODS: Human endothelial cells were isolated from umbilical cord blood. Medium Z-MCF-7-EC was established and was used to co-culture the normal endothelium cells (ECs) and human breast cancer cells of the line MCF-7. Normal endothelial cells cultured by itself were used as control. Light microscopy and transmission electron microscopy were used to observe the morphology of the 2 kinds of endothelial cells. Expression of the genes ESM, IGFBP-3, alphavbeta3, VE C, and Tie-2-2 was analyzed by semi-quantitative RT-PCR using a house-keeping gene beta(2m) as inner reference gene. RESULTS: The ECs co-cultured with MCF-7 breast cancer cells were abnormal in shape with increased size of nucleus and nucleolus, increased size ratio of nuclear to nucleoplasm, increased depth of surface fenestration, loosed and distorted endoplasm, increased size of cell-cell junctions, decreased number of surface microvilli, and tubules formed by ECs. The expression values of the genes ESM, IGFBP-3, alphavbeta3, VE-C, and Tie-2-2 in the ECs co-cultured with MCF-7 breast cancer cells were all up-regulated in comparison with those in the controls (P < 0.01, P < 0.05, P < 0.01, P < 0.01, and P < 0.05 respectively). CONCLUSION: Breast cancer cells promote formation of new vessels with endothelial cells different from the normal ECs in character and behavior. PMID- 16251076 TI - [Influence of elemene on the expression of Bcl-2 family genes in rat C6 glioma cells]. AB - OBJECTIVE: To observe the effects of elemene on the induction of apoptosis in rat C6 glioma cells and its influence on expression of Bcl-2 family genes. METHODS: Rat C6 glioma cells were cultured. Elemene of the concentrations of 0, 20, 40, 60, and 80 microg/ml were added for 12, 24, 36, 48, and 72 hours respectively. RT PCR was used to detect the mRNA expression of Bcl-2/Bcl-x/1 genes. Western blotting was used to detect the protein expression of Bcl-2/Bcl-x/1 genes. The apoptosis of the cells was examined by flow cytometry. RESULTS: The cell counts of the 20, 40, 60, and 80 microg/ml elemene groups were 536 +/- 9, 375 +/- 10, 246 +/- 9, and 112 +/- 10/visual field respectively, all significantly lower than that of the 0 microg/ml elemene group (all F = 1292.416, P < 0.05) and the apoptotic rates of the 20, 40, 60, and 80 microg/ml elemene groups were (27 +/- 2)%, (29 +/- 4)%, (32 +/- 3)%, and (35 +/- 5)% respectively with an Ap peak. The protein expression of Bcl-2/Bcl-x/l genes was decreased in the elemene groups dose and time-dependently. The expression of Bax protein was decreased in the elemene groups too, however, not dose and time-dependently. CONCLUSION: Apoptosis caused by elemene may be associated with the down-regulation of Bcl-2/Bcl-x/l genes. PMID- 16251077 TI - [Effect of human placenta derived mesenchymal stem cells on cord blood lymphocyte transformation]. AB - OBJECTIVE: To study the effect of human placenta derived mesenchymal stem cells (MSCs) on the immune function of lymphocytes derived from human umbilical cord blood. METHODS: Mononucleated cells (MNC) were isolated from human placenta tissue perfusate by density gradient fractionation. Individual colonies were selected and cultured. The culture-expanded cells were characterized by immune phenotyping so as to identify the MSCs. The MSCs were cultured under conditions promoting differentiation to osteoblasts or adipocytes. MNCs were isolated from adult peripheral blood and human umbilical cord blood and cultured, then the adherent cells were excluded and the suspended cells, lymphocytes, were inoculated in the culture fluids of MSCs of different concentrations and phytohemagglutinin (PHA), a nonspecific mitogenic stimulant, was added for 84 hours (MSC + PHA groups), then (3)H-thymidine deoxyribose ((3)H-TdR) was added for 12 hours. The cells were collected and scintillation counter was used to calculate the counts per minute (cpm). Pure lymphocytes without MSC and stimulated by PHA were used as control group (non-MSC Group) and pure lymphocytes and pure MSCs without PHA were used as blank control groups (non-PHA Group). RESULTS: From human placenta MSCs were successfully isolated and exhibited fibroblast-like morphology. Flow cytometric analysis showed that the placental MSCs were a homogeneous cell population devoid of hematopoietic cells positive for CD29, CD44, CD73, CD105, CD166, and HLA-ABC positive and negative for CD34, CD45, and HLA-DR. They could be induced into adipocytes or osteocytes. The cpm value of the non-MSC Group was 171 855 +/- 31 454, significantly higher than that of non-PHA Group (26 453 +/- 5268). The cpm values of the different concentrations MSC + PHA groups were all significantly lower than that of non-MSC Group in a dose-dependent manner; when the dose of MSCs was 2 x 10(5) the suppression rate was 79.97% in PB and 64.06% in UCB. CONCLUSION: MSCs derived from postpartum human placenta, an important and novel source of multipotent stem cells, suppress blood lymphocyte proliferation, thus may be used to reduce graft versus-host disease (GVHD) in recipients. PMID- 16251079 TI - [Effects of testis murine cytomegalovirus infection on sperm acrosome reaction and spermatic function of membrane in mice]. AB - OBJECTIVE: To explore the effects of testis murine cytomegalovirus (MCMV) infection on mature sperm acrosome reaction and spermatic function of membrane in mice. METHODS: BALB/c mice without MCMV infection were randomly devided into two groups: an experimental group (48 mice) and a control group (30 mice). The mice in the control group were treated by inoculation DMEM without MCMV into testis, while those in the experimental one were directly inoculated with MCMV into testis. Mice in two groups were sacrificed separately at 1, 2, 4, 6, 9, 14 d post inoculation (D1, 2, 4, 6, 9, 14 PI), and the MCMV M83 mRNA gene was detected inside the testes by in situ hybridization (ISH) with one episode MCMV late-mRNA probe labeled with digoxin, meanwhile acrosome reaction and the function of membrane of mature sperms in the epididymis tails was measured. RESULTS: The positive signal of ISH of MCMV was mainly founded in the two kinds of testicular cells (spermatogenic cells and Leydig cells) in the experimental group. Compared with that of the control group, the sperm acrosome reaction in the experimental group was decreased significantly by the rate from (71 +/- 6)%, (70 +/- 7)% to (58 +/- 9)%, (56 +/- 9)% (P < 0.05) separately on D2 PI and D4 PI. And the sperm membrane hypo-osmotic swelling was decreased significantly by the rate from (60 +/- 7)%, (50 +/- 4)% to (48 +/- 9)%, (38 +/- 8)% (P < 0.05) separately also on D2 PI and D4 PI. CONCLUSION: The model of CMV infection in murine testis was established. The sperm acrosome reaction and function of membrane in mice might be descented significantly by MCMV infection in the early period, which shows that MCMV infection might influent the sperm's function. PMID- 16251078 TI - [The effect of posttreatment with isoflurane versus propofol on pulmonary alveolar capillary barrier in endotoxemic rats]. AB - OBJECTIVE: To compare the effects of posttreatment with isoflurane versus propofol on pulmonary alveolar capillary barrier (PACB) in endotoxemic rats. METHODS: Ninety-six male Sprague-Dawley rats weighing 250 to 350 g were randomly divided into 6 equal groups to be administered with intravenous lipopolysaccharide (LPS, 4 mg/kg) or equivalent volume of normal saline (NS) and treated with intravenous pentobarbital (C, 20 mg x kg(-1) x h(-1)), inhaled isoflurane (I, 0.55%) or intravenous propofol (P, 10 mg/kg bolus, 40 mg x kg(-1) x h(-1)) 2 hours after the establishment of endotoxemic model: NS-C, NS-I, NS-P, LPS-C, LPS-I, and LPS-P groups. The rats were ventilated for 2 h. The mean arterial pressure (MAP) and arterial blood gas were assessed hourly for 4 h. One cohort (n = 8 per group) was subjected to bronchial alveolar lavage in the left lungs for determination of protein concentration in the bronchial alveolar lavage fluid (BALF) and the lung permeability index (LPI, BALF protein concentration/serum protein concentration). Then the right lungs were removed to undergo light and electron microscopy and measurement of the wet wet-to-dry weight ratio (WW/WD). In a second cohort (n = 8 per group), Evans blue dye (EBD, 30 mg/kg) was injected 30 min before the end of the experiment to determine the EBD content in the lung. RESULTS: The MAP, pH, PaCO(2), and PaO(2) were not different significantly within the three NS groups and the three LPS groups and WW/WD did not show any difference among all six groups. The LPI and EBD content in lung were significantly higher in the LPS-C group [8.1 x 10(-3) +/- 2.4 x 10( 3) and 0.628 +/- 0.082 absorption unit (AU) respectively] than in NS-C group (3.3 x 10(-3) +/- 2.2 x 10(-3) and 0.479 +/- 0.154 AU respectively, both P < 0.05), and were higher in the LPS-P group (9.1 x 10(-3) +/- 2.2 x 10(-3) and 0.664 +/- 0.028 AU respectively) than in the LPS-I group (5.5 x 10(-3) +/- 2.0 x 10(-3) and 0.517 +/- 0.048 AU respectively, both P < 0.05). Light microscopy showed more severe lung injury changes in the LPS groups than in the NS groups, especially in the LPS-P group. Electron microscopy found that the tight junctions between the adjacent pulmonary microvascular endothelial cells in the LPS-P groups were disrupted. CONCLUSION: Compared with posttreatment with propofol, posttreatment with isoflurane protects the PACB function and decreases the pulmonary permeability in endotoxemic rats. PMID- 16251087 TI - [Quality and documentation prior to dermatological laser treatment]. PMID- 16251088 TI - [Children hospitalised with fever following a stay abroad]. AB - INTRODUCTION: The aim of this study was to describe cases of illness in children admitted to hospital with a fever following a stay abroad. MATERIALS AND METHODS: We performed a retrospective study based on journals dating from June 2003 to September 2003. Patients with a temperature of > or = 37.5 degrees C and of whom it was recorded that they had travelled abroad within the previous three months were included, as were demographic data, travel destination, discharge diagnosis and the results of microbiological and serological tests performed, if any. RESULTS: A total of 48 children were included; 56% were immigrants. Fifteen (31%) suffered from diarrhea/gastroenteritis; eight of these cases were caused by Salmonella species. Ten (21%) had a serologically verified hepatitis A infection. There was one case of verified malaria. Nine patients (19%) were discharged with a diagnosis of unspecified viral infection, while nine had a commonly occurring infection with an unidentified focus. CONCLUSION: Immigrant children constituted the majority of the hospitalised children. This group contracts not only more infections but also more serious ones than do children with Danish parents. As for infectious hepatitis, it is remarkable that despite the recommendations of the National Board of Health to vaccinate immigrant children against hepatitis A before a visit to their native country, this seems to be far from the actual practice. PMID- 16251089 TI - [Traveller's diarrhea]. AB - Traveller's diarrhea (TD) is the health problem most frequently encountered by tourists. Neither pretravel advice nor probiotica has shown a consistent prophylactic effect. Antibiotic chemoprophylaxis reduces the incidence of TD by 80-90% but should be recommended only to certain high-risk groups. Vaccination induces only short and partial protection. Standby treatment of TD with rehydration, loperamide and a flouroquinolone is recommended for travellers to most of the high-risk areas of the world. PMID- 16251090 TI - [Imported diseases]. AB - In Denmark, about one third of notifiable diseases are imported, but these have had only a limited spread within the Danish community. For example, imported strains of Mycobacterium tuberculosis remain confined to the immigrant population, and the high prevalence of chronic hepatitis B among certain groups of immigrants has not resulted in a general increase in the incidence of hepatitis B. Exceptions are imported cases of hepatitis A in children, which have resulted in outbreaks in the indigenous Danish population, and the import of drug resistant bacteria, which often pose a serious challenge to hospital hygiene. PMID- 16251091 TI - [Flavivirus infections: yellow fever, dengue fever and Japanese encephalitis]. AB - Flavivirus infections, such as dengue fever, yellow fever and Japanese encephalitis, are untreatable. As a result of the high prevalence of dengue fever in endemic areas, it poses a substantial risk for travellers to those areas. When it comes to Japanese encephalitis and yellow fever, the risk for travellers is limited but the diseases are extremely serious, creating a dilemma for the physician when it comes to deciding whether to vaccinate against them. The vaccines against both potentially have side effects. The indications and contraindications for vaccination are described in detail. PMID- 16251092 TI - [Cerebral malaria after a two-week charter tour to Gambia]. AB - Cerebral malaria is a potentially life-threatening complication of Plasmodium falciparum malaria. We describe a case of severe cerebral malaria in a previously healthy 63-year-old Danish female who had returned from a two-week tourist tour to Gambia. The severe course of the infection described in this case draws attention to two major aspects regarding imported cases of malaria in non-endemic countries: the need for thorough counselling, including chemoprophylaxis, and continuous focus on potential malaria infection after travelling to malaria endemic areas. PMID- 16251093 TI - [Ineffective change of antimalaria prophylaxis to Artemisia vulgaris in a group travelling to West Africa]. AB - Sixteen travellers to West Africa used four kinds of antimalaria chemoprophylaxis. Suspected malaria in three persons and vaginal candidiasis in one caused all seven doxycycline users to change their medication. One of these was persuaded to use Artemisia vulgaris extract. In the course of the three-month journey, there were seven suspected cases of malaria, only two of which could be confirmed by antibody and antigen detection or expert microscopy; both were in travellers who had used A. vulgaris. A. vulgaris had no effect on parasite growth in vitro. The use of natural products for malaria prophylaxis should be discouraged. PMID- 16251095 TI - [Dermatologic problems arising during foreign travel]. AB - The skin is a highly exposed organ during vacation times, especially during travel to countries with subtropical and tropical climates. Prolonged stay in these countries significantly increases the risk of contracting rarely seen dermatoses such as leishmaniasis, larva migrans and myiasis. The bites of various flies may provoke itching and excoriations that may be infected with Staphylococcus aureus and/or hemolytic streptococci, resulting in impetigo, furunculosis or erysipelas. Elderly persons spending weeks in the tropical sun may develop drug-induced phototoxic or photoallergic rash due to concomitant medication for cardiovascular or rheumatic diseases. Acute sunburn is considered a short-lasting problem, but in children it increases the risk of malignant melanoma in later years. Also of concern is chronic UV exposure, which increases the risk of premalignant and malignant skin tumors. Finally, mucocutaneous manifestations arising weeks and months after returning from vacation should raise suspicions of sexually transmitted syphilis and HIV. PMID- 16251094 TI - [Ocular complications of dengue fever]. AB - We report a case of a 32-year-old Danish woman with ocular complications of (IgM verified) dengue fever. After having returned from a trip to Thailand, she became ill with fever and symptoms suggestive of dengue. On the tenth day, when the fever had subsided, she suddenly experienced visual disturbances and was admitted to hospital. Ophthalmologic examination revealed retinal oedema, probably immunologically mediated. Months later, her visual symptoms had still not completely disappeared. Ocular complications of dengue are rarely reported, but in the Third World they are probably common. PMID- 16251096 TI - [Marburg haemorrhagic fever in Angola, 2005]. AB - The current outbreak of Marburg haemorrhagic fever (MHF) in Angola is the largest recorded to date. The article provides an overview of MHF, including historical and clinical aspects, and summarises the status of the present outbreak. Until now, the main components of the public health response in Angola have been effective diagnosis and isolation of patients and contacts, and implementation of hygiene procedures by health workers at local health facilities. Involvement of the local community still constitutes the main challenge for successful control of MHF in Angola. PMID- 16251098 TI - [Potential risks of non-physicians' use of lasers and intense pulsed light in dermatology]. AB - Lasers and light sources are increasingly used by non-physicians, which raises major concern due to the potentially increased risk of errors and complications. Safe treatments depend on optimal preoperative information, examination, and patient selection; optimal performance of the treatments; and optimal postoperative care of wounds and possible complications. Non-physicians do not normally possess these required skills. There is a need for quality assurance in cutaneous laser surgery. This report was initiated by the Danish National Board of Health. PMID- 16251097 TI - [Free versus non-free treatments with laser and intense pulsed light in dermatology: distinguishing medical laser treatments to be provided free of charge from cosmetic self-payment treatments]. AB - Lasers and light sources are increasingly used in dermatology. Due to the limited financial resources of the public health care system, the Danish Dermatological Society has developed recommendations for distinguishing medical laser treatments to be provided free of charge from cosmetic self-payment treatments. Several considerations underlie the recommendations: present legislation; the diagnosis, etiology, severity, and anatomical location of the disease; and the availability of evidence-based treatments. It is suggested that the recommendations constitute the basis for referring patients to dermatological treatment involving laser and intense pulsed light in Denmark. PMID- 16251099 TI - [Picture of the month: thrombus in acute myocardial infarction]. PMID- 16251105 TI - [Commercialization of the medical research is a reality. What is the limit?]. PMID- 16251117 TI - Enzyme-assisted virectomy in enucleated pig eyes. PMID- 16251119 TI - Participation of p38 MAP kinase, but not p44/42 MAP kinase, in stimulation of corneal epithelial migration by substance P and IGF-1. AB - PURPOSE: Substance P and insulin-like growth factor-1 (IGF-1) synergistically promote corneal epithelial migration both in vitro and in vivo. The mechanism of this action was investigated. METHODS: The effects of various inhibitors and activators of intracellular signaling pathways on corneal epithelial migration were examined by measuring the length of the migration path in rabbit corneal blocks in culture. RESULTS: Inhibitors of signaling by p38 or p44/42 isoforms of mitogen-activated protein (MAP) kinase or of phosphatidylinositol (PI) 3-kinase reduced the extent of spontaneous migration of the corneal epithelium, whereas modulators of signaling by cyclic AMP- or cyclic GMP-dependent protein kinases had no effect. The inhibitors of p38 MAP kinase and of PI 3-kinase also abolished the stimulatory effect of substance P and IGF-1 on epithelial migration, whereas inhibitors of signaling by p44/42 MAP kinase or modulators of cyclic nucleotide dependent signaling did not. CONCLUSIONS: These results suggest that various signal transduction systems participate in spontaneous corneal epithelial migration as well as in the combined effect of substance P and IGF-1 on this process. In particular, although both p38 and p44/42 isoforms of MAP kinase appear to regulate spontaneous corneal epithelial migration, the stimulatory effect of substance P and IGF-1 appears to be mediated by p38 MAP kinase but not by p44/42 MAP kinase. PMID- 16251120 TI - N-acetyltransferase 2 phenotype may be associated with susceptibility to age related cataract. AB - Free radicals and oxidative damage play roles in aging and age-related ocular diseases such as cataracts, so defensive mechanisms become important factors for protection. Because N-acetylation is involved in a wide variety of detoxification processes, this study was conducted to examine the relationship between the acetylator phenotypes and genotypes in a group of patients with age-related cataract. Sixty-one cases of age-related cataract and 104 controls were included in this study. Blood was collected in EDTA-containing tubes, and genomic DNA was extracted from the white blood cells by high pure PCR template preparation kit. Genotyping of NAT2 polymorphisms were detected by using a LightCycler-NAT2 mutation detection kit in real-time PCR. There was a significant difference in the distribution of the NAT2*6A acetylator phenotype between cases and the controls. The odds ratio of cataract for the NAT2*6A slow phenotype was 3.8 (95% CI = 1.08 to 13.11, p = 0.032) compared with the fast type. Our results suggest that slow acetylators are at higher risk of developing age-related cataracts than fast acetylators. As NAT2 is an important xenobiotic-metabolizing enzyme and theoretically xenobiotics such as ultraviolet B radiation, smoking, and alcohol use may induce cataract formation, NAT2 gene polymorphisms may be associated with genetic susceptibility of cataract. PMID- 16251121 TI - A mathematical model for ocular tear and solute balance. AB - PURPOSE: In this paper we develop a mathematical model that can predict the steady-state tear film thickness and the dynamic tear film thickness and the solute concentration after instillation of a solute-laden fluid in the eye. METHODS: The mathematical model developed in this paper is based on a balance between the inflow and outflow of tears into the tear film. It incorporates a tear drainage model and a model that relates the tear film thickness to the meniscus radius of curvature. To predict the solute concentrations, the tear balance is coupled with the solute balance. The differential equations for the unsteady balances are solved numerically. RESULTS: The model predicts that the tear film thickness depends on a number of physiological factors, such as rates of tear production and evaporation, geometry and modulus of the canaliculi, and surface tension and viscosity of tears, and varies from about 3 to 15 microm. The model also predicts that the drainage time for an instilled volume of 15 microl is 1283 s. Additionally, the time required for the tracer concentration to decay to 1% of the value immediately after instillation of a drug-laden 40 microl drop is about 2480 s. Similarly, the time for intensity decay for a radioactive tracer after 25 microl instillation is about 1566 s. Also, the model predicts that the fraction of the instilled drug that reaches the cornea is about 1.3% for topical application of timolol. CONCLUSIONS: The predicted results agree reasonably with the reported experimental results, at least qualitatively. The model developed here can serve as a useful tool to develop a more precise understanding of various issues related to tears and also evaluate the effect of various parameters on the tear volume. PMID- 16251122 TI - Morphological changes in the anterior segment of the Abyssinian cat eye with hereditary rod-cone degeneration. AB - PURPOSE: The purpose of this study was to investigate morphological changes of the anterior segment of the eye in Abyssinian cats with progressive rod-cone degeneration and to correlate them with blood flow data obtained in the same animals. METHODS: Sections of the left eyes of six normal cats and of eight cats with different stages of hereditary retinal degeneration were prepared for transmission and scanning electron microscopy. Tangential and sagittal sections were also stained with antibodies against substance P, tyrosine hydroxylase, panneuronal marker PGP9.5, nitric oxide synthase, synaptophysin, and smooth muscle alpha-actin. RESULTS: In Abyssinian cats with hereditary rod-cone degeneration, significant changes were observed in the iris consisting of irregularities in the vascular wall of smaller arteries without changes in their innervation pattern. The ciliary processes were shorter than in normal cats, and their structure appeared more compact and retracted. Slight changes were also observed in the anterior part of the ciliary epithelium. The anterior chamber angle region did not appear to be affected. CONCLUSIONS: Clear morphological correlations to the physiological blood flow data were observed in the anterior eye segment, pointing not only to functional but also morphological vascular abnormalities in this animal model for retinitis pigmentosa. PMID- 16251123 TI - Susceptibility of human corneal endothelial cells to HSV-1 infection. AB - PURPOSE: The purpose of this study was to examine the intrinsic susceptibility of cultured human corneal endothelial cells (HCEC) to herpes simplex virus-1 (HSV-1) infection. We compared HSV-1 adsorption, kinetics of HSV-1 production, and pattern of viral plaque formation in cultured HCEC with those of a cell line used routinely for laboratory HSV propagation (African green monkey kidney fibroblast CV-1 cells). METHODS: Cultured HCEC and CV-1 cells were exposed to the McKrae strain of HSV-1 at 5 and 0.0001 multiplicities of infection (MOI). Using the 50% tissue culture infectious dose (TCID(50)) titration method, viral adsorption (at 5 MOI) and total virus production (at 5 and 0.0001 MOI) were compared to assess both susceptibility to viral attachment and productive viral infection, respectively. Additionally, visual observations were made at 0.0001 MOI using bright-field microscopy and immunofluorescence staining of viral antigens to compare patterns of viral spread in confluent monolayers of both cell types. RESULTS: The percentage of HSV-1 virion particles adsorbed by cultured HCEC and CV-1 cells was similar (35.9% and 33.0%, respectively, p = 0.07, NS), indicating similar susceptibility of the two cell types to initial HSV-1 attachment and adsorption. However, maximum total virus production was more than 3-fold higher for HCEC than for CV-1 cells (p < 0.005), suggesting higher susceptibility of HCEC cells to productive viral infection. Immunofluorescence studies of infected cell monolayers corroborated these quantitative findings, with HCEC monolayers demonstrating more rapid progression of cytopathic effect than CV-1 monolayers. CONCLUSIONS: In comparison to reference CV-1 cells, cultured HCEC show similar susceptibility to HSV-1 adsorption, but higher capacity to support productive HSV 1 infection. Our results suggest that human corneal endothelial cells may be inherently susceptible to HSV-1 infection. PMID- 16251124 TI - Effect of latanoprost on cultured porcine corneal stromal cells. AB - PURPOSE: Latanoprost reduces intraocular pressure mainly by enhancing uveoscleral outflow that may be involved in the decreased of extracellular matrixes such as collagens. However, the effect of latanoprost on corneal stromal cells is not well understood. In the current study, we investigated the changes of cultured porcine corneal stromal cells upon exposure to latanoprost. METHODS: Porcine corneal stromal cells were acquired from primary culture and maintained in fetal bovine serum-containing medium. Cells were estimated on 3H-thymidine, 3H-leucine, 3H-uridine, 3H-proline uptakes and migration. Dead and living cells were estimated with MTT assay. The changes of type 1 collagen and fibronectin proteins were detected by means of immunofluorescent staining and Western blot assay. Intracellular free Ca2+ ([Ca2+]i) mobility was studied by spectrofluorophotometer after loading with fura-2-AM. RESULTS: Latanoprost has remarkable effects inhibiting cultured corneal stromal cells on 3H-thymidine, 3H-leucine, 3H uridine, 3H-proline uptakes and cellular migration. The inhibitory effects are in a dose-dependent manner at concentrations ranging from 10(- 5), 10(- 6), 10(- 7) to 10(- 8) M. The 50% inhibitory dosages (ID50) for latanoprost to corneal stromal cells, as measured by 3H-thymidine uptake, 3H-uridine uptake, 3H-leucine uptake, 3H-proline uptakes and cellular migration were 5.01 x 10(- 6) M, 2.81 x 10(- 6) M, 2.09 x 10(- 6) M, 3.89 x 10(- 7) M and 2.2 x 10(- 6) M, respectively. In the presence of latanoprost, the cellular MTT values were also decreased significantly. Immunofluorescent staining displayed that latanoprost changed type 1 collagen distribution in cultured corneal stromal cells. Western blot assay revealed that latanoprost caused cells to decrease in fibronectin protein. In Ca2+-containing buffer, latanoprost induced a significant rise in [Ca2+]i at 10(- 5) and 10(- 6) M. CONCLUSIONS: These results indicate that latanoprost may induce the morphological and biochemical changes in cultured corneal stromal cells. Long term use of latanoprost needs to be carefully monitored for change in corneal stroma. PMID- 16251125 TI - Production of recombinant human microplasminogen and pilot study in inducing posterior vitreous detachment. AB - PURPOSE: To realize the high production of recombinant human microplasminogen (r mPlg) with Pichia pastoris and demonstrate the efficacy of r-mPlg in inducing posterior vitreous detachment (PVD). METHODS: Recombinant plasmid pAO815-3mPlg was constructed and transformed into SMD1168 cells. Positive recombinant clones were selected with MD plate and cultured in BMG medium, then induced in BMM medium. A protein band corresponding to mPlg with molecular mass of 29 kDa was revealed in SDS-PAGE and confirmed by Western blot. Anion-exchange chromatography and plasminogen activity assay kit were used to obtain purified r-mPlg with biological activity. Twenty eyes of freshly slaughtered pigs were divided into 4 groups, 5 eyes in each group. Group 1 served as normal control. Intravitreal injection of 0.1 ml BSS, 1000 IU/0.1 ml recombinant streptokinase (r-SK) and 1000 IU/0.1 ml r-SK plus 3 U/0.1 ml r-mPlg was done respectively to groups 2, 3, and 4. After incubation at 37 degrees C for 60 min, all eyes were processed for light microscopy, scanning electron microscopy (SEM), and transmission electron microscopy (TEM). RESULTS: r-mPlg, which has potential fibrinolytic activity, was successfully obtained with yield of 30 mg/L and purity of 97%. PVD was demonstrated by SEM in group 4 but not in other three groups. The retina and the inner limiting membrane (ILM) were well preserved in all eyes. CONCLUSION: r mPlg, which has potential fibrinolytic activity, can be produced through Pichia pastoris expression system. Three U of r-mPlg combined with 1000 IU r-SK was effective in producing PVD without damaging the retina. PMID- 16251126 TI - Platelet-activating factor in human normal tears. AB - PURPOSE: To measure the concentrations of platelet-activating factor (PAF) and lyso-PAF in tears of human eyes. METHODS: Unilateral tear samples were collected from the conjunctival cul-de-sac of 12 healthy volunteers without any past histories of ocular surface diseases and 10 patients with allergic conjunctivitis (AC) by graduated disposable microcapillaries. C18:0-PAF, C18:0-lyso-PAF, C16:0 PAF, and C16:0-lyso-PAF levels were determined by liquid chromatography-tandem mass spectrometry (LC/MS/MS). RESULTS: The concentrations of C18:0-PAF, C18:0 lyso-PAF, C16:0-PAF, and C16:0-lyso-PAF in tears from healthy volunteers were 0.44 +/- 0.39, 51.7 +/- 63.4, 61.9 +/- 75.9, and 10.7 +/- 14.7 ng/ml, respectively. Higher, but not significantly different, concentrations of all the four kinds of PAF molecules were detected in tears from AC patients. Significant correlations were demonstrated between the concentrations of C18:0-PAF and C18:0 lyso-PAF (r = 0.906; p < 0.01 in normal healthy volunteers and r = 0.939; p < 0.01 in AC patients), and between those of C16:0-PAF and C16:0-lyso-PAF (r = 0.944; p < 0.01 in normal healthy volunteers and r = 0.806; p = 0.015 in AC patients). Moreover, C18:0-PAF concentrations correlated significantly with those of C16:0-PAF (r = 0.885; p < 0.01 in normal healthy volunteers and r = 0.927; p < 0.01 in AC patients), while C18:0-lyso-PAF concentrations correlated significantly with those of C16:0-lyso-PAF (r = 0.972; p < 0.01 in normal healthy volunteers and r = 0.891; p < 0.01 in AC patients). CONCLUSIONS: To our knowledge, this is the first report of the concentrations of different species of PAF (C18:0-PAF, C18:0-lyso-PAF, C16:0-PAF, and C16:0-lyso-PAF) in human tears. PMID- 16251127 TI - Ocular surface and MUC5AC alterations in atopic patients with corneal shield ulcers. AB - PURPOSE: To describe MUC5AC alterations and the ocular surface disorder in atopic patients with or without corneal ulcers. METHODS: Atopic patients' eyes were divided into two groups according to the presence and absence of corneal ulceration. The subjects underwent corneal sensitivity measurements, Schirmer test, tear film break-up time (BUT), fluorescein and Rose Bengal staining of the ocular surface and conjunctival impression cytology and brush cytology. Impression cytology samples underwent PAS and immunohistochemical staining for MUC5AC. Brush cytology specimens underwent evaluation for inflammatory cell expression and quantitative real-time PCR for MUC5AC mRNA expression. The differences related to the tear function and ocular surface examination parameters between patients with and without corneal ulceration and healthy control subjects were studied. In addition, the differences of the study parameters related to ocular surface epithelial health and inflammatory status between patient eyes with atopic keratoconjunctivitis (AKC) and vernal keratoconjunctivitis (VKC) were investigated. RESULTS: The mean corneal sensitivity and BUT values were significantly lower in atopic patients with corneal ulcers, compared to patients without ulcers and controls (p < 0.001). Brush cytology specimens from patients with corneal ulcers revealed significantly higher expression of inflammatory cells compared to patients without ulcers and controls (p < 0.001). Impression cytology samples from eyes with corneal ulcers showed significant squamous metaplasia and reduction in goblet cell density compared to eyes without ulcers and eyes of control subjects. The mean squamous metaplasia grade was significantly higher in eyes with AKC compared to eyes with VKC (p < 0.02). The mean goblet cell density was significantly lower in eyes with AKC compared to eyes with VKC (p < 0.01). Specimens from eyes with corneal ulcers showed PAS positive mucin pickup and did not stain positive for MUC5AC. MUC5AC mRNA expression was significantly lower in eyes with corneal ulcers compared to eyes without ulcers and eyes of control subjects. MUC5AC mRNA expression was also significantly lower in eyes with AKC compared to eyes with VKC. CONCLUSIONS: Ocular surface inflammation, tear film instability, and decreased conjunctival MUC5AC mRNA expression were thought to be important in the pathogenesis of noninfectious corneal shield ulcers in atopic ocular surface disease. PMID- 16251128 TI - Intraocular metabolites of isopropyl unoprostone. AB - PURPOSE: It is still unknown which metabolite of isopropyl unoprostone is responsible for reducing intraocular pressure. This study was carried out to measure intraocular metabolites of isopropyl unoprostone in the aqueous humor of primate and human eyes. METHODS: Nine monkeys were randomly divided into three groups, all of which received isopropyl unoprostone. In group I, the drug was scheduled to be instilled at 0 hr, in group II at 1 hr, and in group III at 2 hr, prior to aqueous humor aspiration in order to determine metabolite concentration. Furthermore, 27 patients scheduled for cataract surgery and intraocular lens implantation were divided into five groups that received isopropyl unoprostone. In group A, the drug was scheduled to be instilled at 0 hr, in group B at 1 hr, in group C at 2 hr, in group D at 3 hr, and in group E at 4 hr, prior to surgery. At the beginning of the operation, the aqueous humor was aspirated. Metabolites of isopropyl unoprostone in the aqueous humor were determined by high-performance liquid chromatography. RESULTS: M1 (3-[(1R,2R,3R,5S)-3,5-dihydroxy-2-(3 oxodecyl)cyclopentyl]propionic acid) (unoprostone free acid) and M2 ((Z)-7 [(1R,2R,3R,5S)-3,5-dihydroxy-2-(3-oxodecyl)cyclopentyl]hept-5-enoic acid), an intraocular oxidized metabolite of isopropyl unoprostone, were measured. M1:M2 in monkeys was respectively 0:0 ng/ml in group I, 150.2 +/- 45.1:9.5 +/- 1.7 (p < 0.05) in group II, and 74.6 +/- 31.4:19.2 +/- 5.3 (p < 0.01) in group III. M1:M2 in humans was respectively 0:0 ng/ml in group A, 50.6 +/- 22.3:3.2 +/- 1.3 (p < 0.05) in group B, 125.0 +/- 23.1:12.2 +/- 3.4 (p < 0.001) in group C, 144.9 +/- 33.8:24.5 +/- 6.2 (p < 0.01) in group D, and 56.7 +/- 21.5:18.7 +/- 5.3 (p < 0.05) in group E. CONCLUSIONS: A free acid of isopropyl unoprostone is the major intraocular metabolite of isopropyl unoprostone that is expected to act on target tissues in the eyes of both monkeys and humans. PMID- 16251130 TI - Glutathione reductase from human cataract lenses can be revived by reducing agents and by a molecular chaperone, alpha-crystallin. AB - PURPOSE: The aim of this study was to investigate how glutathione reductase (GR) loses its activity during cataract formation and whether it is possible to revive it back to the normal levels. METHOD: In this study, endogenous as well as synthetic reducing systems (GSH, TTase, DTT, captopril) and alpha-crystallin at different concentrations were incubated with the soluble fraction of human cataract lens protein. The activity of glutathione reductase with or without the reducing agents and alpha-crystallin was tested, and the difference in activity gained was calculated. RESULTS: Five agents (GSH, DTT, TTase, captopril, alpha low crystallin) were able to revive the activity of GR from human cataract lenses to different extents. CONCLUSION: This study shows that human lens GR activity was revived by different reducing agents as well as by a molecular chaperone (alpha-crystallin). PMID- 16251129 TI - Effects of dehydration on corneal tissue absorption of topical azithromycin in rabbits. AB - PURPOSE: To assess the corneal tissue absorption of azithromycin in desiccated and normal eyes of rabbits. METHODS: A total of 25 New Zealand Albino rabbits weighing 2-3 kg each were included. One eye of each rabbit was desiccated. The other eye was left as control. Azithromycin 1.5% topical eyedrops were instilled in both eyes. Five rabbits were sacrificed at each of the following time points: 30 min, 3 hr, 6 hr, and 12 hr. Corneal tissues specimens were subjected to high performance liquid chromatography mass spectrometry. A paired t test was used to evaluate the statistical difference in corneal tissue absorptions of azithromycin at each time point. RESULTS: The mean corneal tissue levels of azithromycin in dry eyes were 66.3 microg/ml, 92.6 microg/ml, 117.5 microg/ml, and 179.9 microg/ml, and the mean corneal tissue levels of azithromycin in normal eyes were 42.0 microg/ml, 43.4 microg/ml, 43.3 microg/ml, and 80.0 microg/ml at 30 min, 3 hr, 6 hr, and 12 hr respectively. Both groups showed increase in corneal tissue absorption overtime (p < 0.0001). Significantly higher levels of azithromycin were noted in dry eyes at each time point except at the 30-min time point. CONCLUSIONS: This study demonstrated that corneas exposed to desiccation showed statistically significant increase in azithromycin level compared to normal eyes. PMID- 16251131 TI - Wayfinding in familiar and unfamiliar environments in a case of progressive topographical agnosia. AB - A 71-year-old right-handed man (F.G.) presents with prosopagnosia and with an inability to recognize famous and familiar buildings. Despite his deficit, F.G. obtained normal scores on neuropsychological tests of executive functions, language, praxis and primary visuoperceptual skills. Brain MRI showed atrophy predominantly in the right temporal lobe, particularly in the fusiform gyrus and the parahippocampal cortex. The present study investigated F.G.'s ability to orient himself in familiar and new environments. His wayfinding abilities in a familiar environment (i.e., his hometown) were preserved despite an inability to recognize familiar and famous buildings, monuments and landmarks in this environment. Wayfinding was achieved through a heavy reliance on written indications (e.g., names of restaurants and streets), preservation of a pre existing cognitive map of this familiar environment, and normal executive functions necessary to plan the execution of a given trajectory. In an unfamiliar environment, F.G.'s topographical agnosia resulted in severe wayfinding difficulties and in the inability to build an adequate cognitive spatial representation. F.G.'s topographical agnosia results from a high-level visuoperceptual deficit, characterized by an inability to access a global configuration of complex visual stimuli such as familiar and famous monuments, and an over-reliance on the processing of local features. PMID- 16251132 TI - Left hemispatial visual neglect associated with a combined right occipital and splenial lesion: another disconnection syndrome. AB - Damage to the left occipital lobe and the splenium or forceps major is often associated with pure alexia, thought to be an occipital-temporoparietal disconnection syndrome. A patient with the parallel lesion, a combined right occipital and splenial lesion, showed severe left-sided visual spatial neglect, but no significant neglect in other sensory modalities. This visual neglect might be related to a disconnection between the visual information processed by the left occipital lobe and the right posterior temporal-inferior parietal areas that mediate attention in the left hemispace. PMID- 16251133 TI - Brain damage and cortical compensation in foreign accent syndrome. AB - Foreign Accent Syndrome (FAS) is a well-known neurological deficit whose underlying cause has remained obscure despite almost a century of study. Combining structural and functional imaging, our studies suggest that FAS represents a compensatory response to impaired motor regulation of speech. We describe a patient who acquired FAS as a result of an ischemic stroke in the left basal ganglia. In addition to this case being exceptionally clean, we were able to confirm a specific lesion location as well as provide strong evidence that impaired motor speech regulation resulted in compensation by other areas of the cortical motor speech network. PMID- 16251134 TI - Cognitive and neural mechanisms underlying reading and naming: evidence from letter-by-letter reading and optic aphasia. AB - We report detailed analysis of language performance in a patient, RMI, a 55-year old man who presented with a homonymous hemianopsia, optic aphasia, and alexia without agraphia (with letter-by-letter reading) acutely after stroke. MRI showed infarct in the left occipital and medial temporal lobe and hypoperfusion of the entire posterior cerebral artery territory, including the splenium. Extensive language testing revealed severely impaired picture naming and oral reading, with relatively spared tactile naming and recognition of orally spelled words, consistent with impaired access to lexical and semantic representations from vision. In addition, he had a milder deficit in accessing lexical representations for output from all input modalities. RMI's execution of various language tasks provided considerable insight into the mechanisms that underlie oral reading. His performance indicated that both semantic access and orthographic to phonologic conversion mechanisms were partially intact. When information from these two impoverished systems was coupled (the picture of an object presented with its written name), his ability to read/name improved significantly, consistent with the hypothesis that partially accessed semantic information from vision can combine with partially accessed sublexical orthographic to phonologic conversion mechanisms to access phonological representations for output. Furthermore, his written word and picture recognition improved to normal at a time when magnetic resonance perfusion imaging demonstrated reperfusion of the splenium. We interpret these results, as well as results from previous studies in the literature, within a model of the neural regions critical for various cognitive processes underlying reading. PMID- 16251135 TI - Knowledge of living, nonliving and "sensory quality" categories in semantic dementia. AB - This article reports the findings from 3 patients with semantic dementia (SD) who were given a novel battery of 33 items from sensory quality categories (SQCs) as previously described by Borgo and Shallice (2001; 2003) and Laiacona, Capitani and Caramazza (2003). Their performance on three tasks (two naming, one word-to picture matching) was compared with performance on similar tasks using a conventional semantic battery. At the group level, patients performed worse than age-matched controls overall, but neither group showed any differences in performance between domains (i.e., living, nonliving and SQCs). Individual patient analyses, however, showed contrasting profiles in the three patients. The results are discussed in terms of the SFT (Warrington & Shallice, 1984) and individual differences (Lambon-Ralph et al., 2003) accounts of category specificity in SD. PMID- 16251136 TI - Jargon dyslexia in an individual with semantic dementia: further evidence for task-specificity in phonological output. AB - We report a patient with semantic dementia who demonstrated a very unusual dyslexia. He had a global loss of conceptual knowledge in the context of a fluent dysphasia and intact syntax. However, he did not have the surface dyslexia which is typical of semantic dementia; rather his reading impairment was characterized by speech production errors resulting in multiple neologisms. In a series of experiments it was established that input phonological and input orthographical processing were intact as was output phonology for naming and propositional speech. We demonstrate that our patient has a task-specific phonological deficit and we argue that reading and propositional speech rely upon dissociable phonological output systems. Thus we corroborate our earlier evidence of task specific phonological output stores (Crutch and Warrington, 2001). We also document a greater difficulty with comprehending the written than the spoken word. We account for this pattern of performance in terms of our patient's attempting to read by the indirect phonological route, as with other semantic dementia patients, but suggest that this process is overridden by the task specific speech production deficit. PMID- 16251137 TI - Synaesthesia: discordant male monozygotic twins. AB - Grapheme-color synaesthesia, a condition in which achromatic graphemes elicit vivid experiences of color is believed to be a genetically determined trait. We describe a study of 10-year-old twin brothers who are physically identical in appearance but who have considerably different conscious experiences. A phenotypic analysis that measured the consistency of grapheme-color pairings over test-retest confirmed that one twin has grapheme-color synaesthesia and the other twin does not. A genotypic analysis using sixteen microsatellite loci confirmed that the twins are monozygotic. These findings are problematic for previous suggestions that synaesthesia is an X-linked dominant trait. At the very least, the findings show that the penetrance of the genotype for synaesthesia is incomplete and that any view suggesting that synaesthesia is simply an X-linked dominant trait is therefore also incomplete and possibly even incorrect. The findings also negate a previous suggestion, based on a study of female monozygotic twins, that discordance of synaesthesia in identical twins is due to X-inactivation. In general, the findings raise serious questions regarding whether it is possible at this time to establish the genetic contribution to synaesthesia. PMID- 16251138 TI - Presymptomatic semantic impairment in a case of fronto-temporal lobar degeneration associated with the +16 mutation in MAPT. AB - We describe a patient who came to neurological attention because of his at-risk status for the +16 exon 10 splice mutation in the tau gene (microtubule associated protein tau, MAPT), which had given rise to progressive behavioural disturbances in two of his siblings. The patient began to exhibit early signs of behavioural disturbance at around the age of symptom onset in both of his siblings. Although he did not spontaneously complain of difficulties in the domain of language, he met clinical, radiological and neuropsychological criteria for semantic dementia. On the assumption that his illness is mediated by the same pathological process as those of his siblings, we propose that this clinical picture represents the earliest changes of a semantic impairment - a phase of the illness that is often retrospectively described by patients and their relations, but has never previously been documented at first hand. Although typical of semantic dementia in many respects, the illness had several interesting and atypical features that emerged on detailed testing: first, he exhibited no insight into his difficulties; secondly, progression over a twelve-month interval was unusually slow; thirdly, he evinced a striking and consistent advantage for nonliving over living concepts; fourthly, a differential impairment of distinctive over shared knowledge did not emerge except when items that he could still name were compared with those for which he was anomic. Finally, the availability of post mortem pathological analysis from the brains of both of his affected siblings allowed us to attribute his illness to a specific pathological process which is considered unusual for patients with this clinical phenotype. PMID- 16251139 TI - Short-term motor and sensory outcomes in acquired nonaccommodative esotropia of childhood. AB - Information on the management of acquired nonaccommodative esotropia (ANAET) of childhood is incomplete. Most prior reports combine the results of patients with both accommodative and nonaccommodative esodeviations. The primary objective of this study was to describe the early postoperative motor and sensory results of children with acquired nonaccommodative esotropia. The medical records of 72 consecutive children with ANAET who underwent surgical correction at East Tennessee State University College of Medicine from August 1, 1995 through October 1, 2001 were retrospectively reviewed. The median age at surgery for the 72 study patients was 43.8 months with a median angle of deviation of 30 prism diopters (PD) at both distance and near. Ten patients (13.9%) required a second surgical procedure while one patient (1.4%) required a third. The cumulative probability of a second surgery within two years of the first procedure was 24% (95% CI 4-43%). During a median postoperative follow-up period of 7.1 months, 64 (88.9%) patients were within 8 PD of orthotropia on the last postoperative examination. Normal postoperative stereoacuity was achieved in several children whose deviation began after 30 months of age, as well as in one child who was misaligned for at least two years prior to surgery. Although the postoperative follow-up on these patients with acquired nonaccommodative esotropia managed without prism adaptation is short, the motor results are similar to other studies on patients with acquired esotropia, with sensory outcomes superior to those obtained in children with congenital esotropia. PMID- 16251140 TI - Drift in ocular torsion during sustained head tilt. AB - PURPOSE: A head tilt towards the shoulder (roll) induces an ocular counter-roll (OCR), i.e. torsion in the opposite direction to the head. How this counter rolled position is maintained during a static head tilt is in debate. In a previous study, we reported an OCR-increasing drift subsequent to the head tilt. This finding is in contrast to other reports where no such response was found. The primary aim of this study was to repeat the experiment during a prolonged head-tilt test and to describe the OCR characteristics. A secondary aim was to investigate the influence of spatial visual cues on OCR. METHODS: Five male subjects performed a head tilt (30 degrees ) towards the right shoulder while the eye position was recorded during a 10-minute interval. In test 1, the subjects viewed a target with no cues for spatial orientation. The same head-tilt paradigm was repeated in test 2 with a visual target with spatial cues. Two samples of data were extracted from the start and the end of the recordings for statistical analysis. RESULTS: Subsequent to the head tilt, a slow OCR-increasing drift in the opposite direction to the head roll was found in all subjects. On average, this drift lasted for 30 sec (+/- 5) in test 1 and for 55 sec (+/- 18) in test 2. The drift was then found to change its direction, i.e. the eyes were rotated in the same direction as the head roll. When measured after 10 minutes, the OCR was significantly decreased. CONCLUSIONS: The OCR during static head tilt is not constant. During the first minute there is a gradually increasing OCR. Thereafter, the amplitude of the OCR decreases gradually. These changes are influenced to some extent by spatial visual cues. Possible mechanisms are adaptive responses in otolithic afferents as well as central nervous memory functions related to the semicircular canal system. PMID- 16251141 TI - Asymmetrical vertical phorias in normal subjects: the influence of unbalanced illumination. AB - PURPOSE: One goal of this investigation was to determine whether asymmetrical vertical hetereophorias, which are qualitatively similar to a dissociated vertical deviation (DVD), occur commonly in normal people. A second goal was to establish whether the magnitude or direction of vertical phoria depends on the illumination of the occluded eye, as would be expected if vertical eye position were influenced by a dorsal light reflex. METHODS: Vertical phorias were estimated in 30 normal subjects using a Maddox rod in conjunction with a dark and an illuminated occluder. RESULTS: On average, subjects exhibited a hypophoria of less than 1 min arc with both the dark and the illuminated occluder. Asymmetrical vertical phorias that were consistent with previously published criteria for a minute DVD were found in eight subjects when a dark occluder was used and in four subjects when an illuminated occluder was used. The proportions of subjects whose vertical phorias were consistent with a DVD did not differ significantly under the two conditions of occlusion. CONCLUSIONS: Asymmetrical vertical phorias that mimic a minute DVD do not occur commonly in normal people. Our finding that vertical phorias do not change systematically when an illuminated instead of a dark occluder is used suggests that normal subjects exhibit little or no vestige of the dorsal light reflex. PMID- 16251142 TI - Isolated abducens nerve palsy associated with retinoic acid therapy: a case report. AB - Isolated abducens nerve palsy is a rare complication of treatment with various drugs. Here, the authors report the case of a 23-year-old female with isolated left abducens nerve palsy after long-term retinoic acid therapy. The association is based on the temporal relationship and the exclusion of other possible etiologic factors following extensive laboratory and imaging diagnostics. The authors suggest that isolated abducens nerve palsy may be a presenting sign of a toxic neuropathy associated with retinoic acid therapy. After the exclusion of other organic lesions, especially idiopathic intracranial hypertension, and an assessment of the risk-benefit ratio, discontinuation of treatment must be considered in such cases. PMID- 16251143 TI - Update on chronic progressive external ophthalmoplegia. AB - Mitochondrial encephalomyopathies are clinically and genetically heterogeneous disorders. External ophthalmoplegia is the most frequent symptom. Other frequently involved tissues and organs include the retina, heart, limb muscles, peripheral and central nervous system, inner ear and endocrine system. The diagnosis is based on the finding of elevated serum lactate, the characteristic histopathological changes in the muscle biopsy, and decreased activities of mitochondrial respiratory chain enzymes. In many cases, the underlying molecular defect in the mtDNA can be identified. The efficacy of pharmacological therapies (e.g., coenzyme Q) has not been established so far. Symptomatic ophthalmological treatment includes ptosis and strabismus surgery. Early cardiac pacemaker implantation may be life-saving. PMID- 16251144 TI - Charles de Saint-Yves (1677-1736). PMID- 16251145 TI - On squint. PMID- 16251146 TI - A description of the Gunn pupil in 1722. PMID- 16251147 TI - Marlow occlusion: does it create or eliminate artifact? PMID- 16251148 TI - Postural control and detection of slip/fall initiation in the elderly population. AB - One of the common causes of morbidity and mortality in workplaces is related to slips or falls. Reaction to external disturbances, such as slips or falls, requires a process of perturbation detection and control of motion changes. Postural control is a common mechanism to compensate unexpected displacements of the body. The ability of postural control diminishes with ageing or neuropathy. In this study, two controlled groups, diabetics and non-diabetics in the elderly population, were investigated to compare how different postural control mechanisms would relate to the detection of perturbation and regain of balance. The ultra-low-vibration Sliding Linear Investigative Platform for Analyzing Lower Limb Stability SYSTEM was used to measure the biomechanical changes of posture and perturbation detection. In phase 1 of the analysis, static measures during quiet standing were considered to investigate the relationship between postural stability and perturbation detection capability. In phase 2 of the analysis, dynamic measures during an occurrence of perturbation were analysed. Statistical tests and linear logistic regression models were applied to find differences of postural control mechanisms and to build a predictive model for perturbation detection quantitatively. It is anticipated that the results of this study will contribute to more comprehensive understanding of postural control mechanisms and design of slip/fall prevention programmes. PMID- 16251149 TI - Psychological impact of air velocity variations in a ventilated room. AB - Two experiments investigated the psychological impact of two velocity conditions (constant low velocity (V1) and variations of low and high velocity (V2)) in two temperature conditions (Experiment 1: an air temperature increase from 21 degrees C to 24 degrees C; Experiment 2: an air temperature increase from 25 degrees C to 27 degrees C) in females and males, aged 16 to 18 years, under realistic classroom conditions during an exposure period of 80 min. It was predicted that the V2 room condition compared to the V1 room condition would be more beneficial for subjects' perceived room temperature and air quality, self-reported affect and cognitive performance. The results obtained showed no significant effects on cognitive performance. However and as predicted, in Experiment 1, the subjects in the V2 compared to those in the V1 room condition felt that the air temperature decreased (while it de facto increased) and reported a constant level of high activation. In Experiment 2, the subjects in the V2 room condition felt that the air temperature increased less and reported that their unactivated unpleasantness increased less and activated pleasantness decreased less than it did for subjects in the V1 room condition. All this indicates, as was suggested by Wigo et al. (2002), that a cooling effect, induced by air velocity variations, might be beneficial for subjects in a ventilated room and that their perceived pleasantness of the indoor climate could be met at a higher room temperature than otherwise. PMID- 16251150 TI - Validating task analysis for error identification: reliability and validity of a human error prediction technique. AB - This paper reports on the theoretical and empirical developments for an error prediction methodology called task analysis for error identification (TAFEI). Other researchers have noted the need for theoretically driven approaches that are able to provide practical utility in error prediction. Theoretical developments include the concept of "rewritable routines", which describe the loop between cognitive processing, action and devices states. This has been proposed as a way of unifying ideas from systems theory and cognitive psychology. The empirical research shows that TAFEI is superior to heuristic methods, which supports the idea that structured methods assist in error prediction. The validation study shows that TAFEI reaches acceptable levels in terms of test retest reliability and concurrent validity. It is believed that the method has reached a level of maturity after 10 years of development work. This is demonstrated by the many uses to which the method has been put, including that of a design tool. PMID- 16251151 TI - Visual effects of the luminance surrounding a computer display. AB - The luminance surrounding a computer display can potentially reduce visibility of the display (disability glare), result in sensations of discomfort (discomfort glare) and result in transient adaptation effects from fixating back and forth between the two luminance levels. The study objective was to measure the effects of surround luminance levels upon these functions in younger and older adults to determine recommended surround luminance levels. The younger age group comprised 20 subjects (mean age 27.9 years, range 23 - 39) and the older group 17 subjects (mean age 55.5 years, range 47 - 63). The central task was presented with luminance of 91 cd/M(2), tested surround luminance levels were 1.4, 2.4, 8.9, 25.5, 50, 91, 175, 317, and 600 cd/M(2). Disability glare was tested with low contrast (20%) visual acuity charts, transient adaptation was tested with a task that required regular fixation between the two luminance levels, discomfort was measured with a questionnaire after reading stories with different surround luminance levels, and preferred luminance was measured by method of adjustment. The surround luminance significantly affected transient adaptation (p < 0.0001), optimal performance occurred at 50 cd/M(2) and above for the young group and at 91 cd/M(2) and above for the older group. Neither low contrast acuity (disability glare) nor symptoms when reading were significantly affected by surround luminance. There was wide variation in preferred surround luminance; however, average preferred surround luminance was 86.9 cd/M(2) for the young group and 62.2 cd/M(2) for the older group, slightly below the central luminance of 91 cd/M(2). The effects of the surround luminance within the tested range are not large; however, the data show that the lowest surround luminance levels should be avoided and that surround luminance levels at or slightly below that of the central task are preferred. PMID- 16251152 TI - Multimodal feedback for the acquisition of small targets. AB - This paper examines how multimodal feedback assists small-target acquisition in graphical user interfaces. All combinations of three feedback modes are analysed: non-speech audio; tactile; and pseudo-haptic "sticky" feedback. The tactile conditions used stimulation through vibration (rather than force-feedback), and the sticky conditions were implemented by dynamically reconfiguring mouse control display gain as the cursor entered the target. Results show that for small, discretely located targets all feedback modes reduce targeting times, with stickiness providing substantial improvements. Furthermore, stickiness and tactile appear to combine well. However, the results of a more ecologically oriented menu-selection task show the need for caution, revealing that excessive feedback can damage interaction though "noise" that interferes with the acquisition of neighbouring targets. PMID- 16251153 TI - Toward development of a computer-based methodology for evaluating and reducing medication administration errors. AB - The number of people aged 65 years and older in the United States is increasing. This age group consumes 30% of all prescription drugs. Many elderly people require assistance from caregivers in taking their medication. Medication administration errors can result if caregivers cannot remember to give the medications or do not understand how to give them. This study aims to determine a more effective method of presenting prescription instructions and to determine if the multiple resource hypothesis holds in the context of prescription instructions by evaluating the effect a voice prescription label (which gives audio instructions) has on comprehension and memory of a drug regimen under varying training level, task condition and instruction format. In performing a multivariate ANOVA on data collected among 48 formal and 48 informal caregivers, training level, task condition, sound condition and instruction format were found to significantly affect caregivers' memory and comprehension. There is evidence that audio instructions and the matrix format reduce errors. These results could lead to the development of a medication scheduling management SYSTEM that effectively provides prescription instructions, organizes medicines according to administration time and incorporates decision rules to determine what action should be taken if a dose is missed. PMID- 16251154 TI - Available friction of ladder shoes and slip potential for climbing on a straight ladder. AB - Straight ladder accidents are a major safety problem. As a leading cause of injuries involving straight ladders, slips at the ladder base occur when the required friction exceeds the available friction at the ladder shoe and floor interface. The objectives of this experiment were to measure the available friction at the base of a portable straight ladder in contact with a floor and to estimate the slip potential of the ladder. The results of friction measurements indicated that the measured friction coefficient on the oily surfaces differed among the six commercially available ladder shoes evaluated. A statistical model was used to compare the available friction results from the current study with the friction requirements under different climbing conditions from a previous study based on their stochastic distributions to estimate the slip potential at the base of the ladder. The results showed that different climbing conditions used in the previous study could be supported by available friction on dry surfaces. However, when the ladder was put onto oily surfaces, resulting in a significant reduction in the available friction due to contamination, slip potential was significantly increased. PMID- 16251155 TI - Reducing whole-body vibration and musculoskeletal injury with a new car seat design. AB - A new car seat design, which allows the back part of the seat (BPS) to lower down while a protruded cushion supports the lumbar spine, was quantitatively tested to determine its effectiveness and potentials in reducing whole-body vibration (WBV) and musculoskeletal disorders in automobile drivers. Nine subjects were tested to drive with the seat in: 1) the conventional seating arrangement (Normal posture); and 2) the new seating design (without BPS (WO-BPS) posture). By reducing contact between the seat and the ischial tuberosities (ITs), the new seating design reduced both contact pressure and amplitude of vibrations transmitted through the body. Root-mean-squared values for acceleration along the z-axis at the lumbar spine and ITs significantly decreased 31.6% (p < 0.01) and 19.8% (p < 0.05), respectively, by using the WO-BPS posture. At the same time, vibration dose values significantly decreased along the z-axis of the lumbar spine and ITs by 43.0% (p < 0.05) and 34.5% (p < 0.01). This reduction in WBV allows more sustained driving than permitted by conventional seating devices, by several hours, before sustaining unacceptable WBV levels. Such seating devices, implemented in large trucks and other high-vibration vehicles, may reduce the risk of WBV-related musculoskeletal disorders among drivers. PMID- 16251160 TI - Differences in parenting stress between married and single first time mothers at six to eight weeks after birth. AB - Parenting stress has been shown to affect relationships between mothers and their infants. Single mothers may experience more parenting stress due to exposure to stressors that are unique to their environment. The purpose of this secondary analysis was to determine the differences between married and single first-time mothers on the Parenting Stress Index/Short Form (PSI/SF Abidin, 1995). A convenience sample of 22 single and 52 married first-time mothers were recruited from postpartum units in two large metropolitan hospitals in the southeastern United States. During hospitalization, participants completed a demographic sheet and at six weeks postpartum, were mailed the PSI/SF. T-tests were assessed for differences between mean scores. Study findings indicate significant differences between single and married first-time mothers on the Parental Distress subscale (t = -3.468, p = .001), the Difficult Child subscale (t = -2.032, p = .046), and Defensive Responding subscale (t = -3.720, p = .000), with single mothers consistently scoring higher than married mothers; however, there was not a significant statistical difference between mothers on the Parent-Child Dysfunctional Interaction subscale (t = -1.290, p = .201). Overall, single mothers scored higher on the Total Parenting Stress scores of the PSI/SF than married mothers (t = -2.858, p = .005), suggesting that single women experience more stress as a new mother than married new mothers. It is important to understand the specific stressors of new, single mothers and to develop nursing interventions that decrease parenting stress in single mothers. PMID- 16251156 TI - Reliable exposure assessment strategies for physical ergonomics stressors in construction and other non-routinized work. AB - The objective of this research was to provide guidelines for the reliable assessment of ergonomics exposures in non-routinized work. Using a discrete interval observational sampling approach, two or three observers collected a total of 5852 observations on tasks performed by three construction trades (iron workers, carpenters and labourers) for periods of several weeks. For each observation, nine exposure variables associated with awkward body postures, tool use and load handling were recorded. The frequency of exposure to each variable was calculated for each worker during each of the tasks on each of the days. ANOVA was used to assess the importance of task in explaining between-worker and within-worker variability in exposures across days. A statistical re-sampling method (bootstrap) was used to evaluate the reliability of exposure estimates for groups of workers performing the same task for different sampling periods. Most exposures were found to vary significantly across construction tasks within trade, and between-worker exposure variability was generally smaller than within worker exposure variability within task. Bootstrapping showed that the reliability of the group estimates exposure for the most variable exposures within task tended to improve as the assessment periods approached 5-6 d, with marginal improvements for longer assessment periods. Reliable group estimates of exposure for the least variable exposures within task were obtained with 1 or 2 d of observation. The results of this study demonstrate that an initial estimate of the important environmental or task sources of exposure variability can be used to develop an efficient sampling strategy that provides reliable estimates of ergonomics exposures during non-routinized work. PMID- 16251162 TI - Further psychometric testing and use of the Maternal Confidence Questionnaire. AB - Research results can be better interpreted and used when reliable and valid instruments are used to measure concepts. Researchers and clinicians, especially those from locations outside the United States, have difficulty locating existing instruments for use in their research studies. The dilemma is further exaggerated when such instruments are not published and when the author of the instrument cannot be located. This review looked at the results of 20 studies which have utilized the Maternal Confidence Questionnaire (MCQ) (Zahr, 1991) with the purpose of familiarizing researchers and clinicians with its flexibility, reliability, and validity. Examples of specific published studies which have used the instrument are discussed along with unpublished studies which have used the instrument. PMID- 16251161 TI - School-age children with asthma and their parents: relationships with health care providers. AB - Health care providers, including nurses, physicians, and other personnel, are key figures who design and implement plans of care to help families manage childhood asthma, yet families' perceptions of relationships with these professionals has received limited study. Child and parent perspectives about relationships with their health care providers emerged as themes in a study that explored responsibility sharing between school-age children with asthma and their parents (Buford, 2004). Fourteen school-age children with asthma and 14 of their parents from 11 families participated in the study. Parents and, to a lesser extent, children, described aspects of their relationships with their health care providers that were supports or barriers to asthma management. Implications for nurses and other health care providers stem from these data and include the importance for health care providers to educate themselves and their patients about state-of-the-art asthma care. Education should be directed to both parents and their children. In addition, parents need to receive education about how to coach their children because the children depend on them for information and direction. Finally, nurses and other health care providers need to listen to parents and value their input about their children's conditions. PMID- 16251163 TI - Eating behaviors among school-age children associated with perceptions of stress. AB - Eating has been theorized to be useful as a coping strategy in response to stressful situations. However, investigation of this behavior in children is limited. The present study is a secondary cross-sectional analysis of longitudinal data that were collected from cohorts of fourth-, fifth-, and sixth grader students. Perceived stress was correlated with unhealthy eating behaviors (r = .13, p < .001), as well as with the use of eating as a coping mechanism (r = .24, p < .001). Hispanic children reported using eating as a coping mechanism most frequently, followed by African-American and Caucasian children. School-age children who experience high levels of stress may be at risk for developing unhealthy eating habits in order to cope; continued examination of these relationships is suggested. Future research should focus on the development of interventions to encourage positive coping mechanisms and healthy eating behaviors. PMID- 16251165 TI - Bipolar depression: issues in diagnosis and treatment. AB - Although bipolar affective disorder is defined by the history of manic or hypomanic episodes, depression is arguably a more important facet of the illness. Depressive episodes, on average, are more numerous and last longer than manic or hypomanic episodes, and most suicides occur during these periods. Misdiagnosis of major depressive disorder delays initiation of appropriate therapy, further worsening prognosis. Distinguishing features of bipolar depression include earlier age of onset, a family history of bipolar disorder, presence of psychotic or reverse neurovegetative features, and antidepressant-induced switching. Bipolar I depressions should initially be treated with a mood stabilizer (carbamazapine, divalproex, lamotrigine, lithium, or an atypical antipsychotic); antidepressant monotherapy is contraindicated. More severe or "breakthrough" episodes often require a concomitant antidepressant, such as bupropion or a selective serotonin reuptake inhibitor (SSRI). The first treatment specifically approved for bipolar depression is a combination of the SSRI fluoxetine and the atypical antipsychotic olanzapine. For refractory depressive episodes, venlafaxine, the monoamine oxidase inhibitor tranylcypromine, and ECT are most widely recommended. The optimal duration of maintenance antidepressant therapy has not been established empirically and, until better evidence-based guidelines are established, should be determined on a case-by-case basis. PMID- 16251166 TI - Psychotherapy and social support: unsettling questions. AB - There is an extensive body of research that documents the importance of social support as an independent factor in physical health and longevity. This article reviews published research that addresses the effects of psychotherapy on social connection and social support. The assumption of most psychotherapists that psychotherapy generally improves social connection is unsupported by research data except in very specific diagnostic categories. Evidence that otherwise beneficial psychotherapy can inadvertently damage social connections is also considered. The implications of these findings for further research and current practice are discussed. PMID- 16251167 TI - A source-monitoring account of auditory verbal hallucinations in patients with schizophrenia. AB - Auditory verbal hallucinations are a common and distressing symptom experienced by patients with schizophrenia. They can be understood as arising from an impairment in reality monitoring-the process by which internally and externally generated events are distinguished. This impairment might arise through primary abnormalities in the reality-monitoring mechanism or through secondary mechanisms (abnormalities in the perceptual characteristics of internally generated events or in the perception of externally generated events). This article examines evidence for and against an association between abnormalities in reality monitoring and auditory verbal hallucinations in schizophrenia. A comprehensive review of the psychological literature suggests that there is little evidence for an association between auditory verbal hallucinations and secondary mechanisms leading to abnormalities in reality monitoring. There is some evidence suggesting that hallucinators show a primary reality-monitoring abnormality that is most apparent when patients are required to distinguish self from other in real time. To draw firmer conclusions, however, it is imperative that future studies select patient populations precisely, match control groups, and use consistent criteria for defining hallucinators. PMID- 16251168 TI - The food preoccupations of a nine-year-old girl: what do they portend? PMID- 16251169 TI - The essentials of neuropsychiatry: teaching residents and fellows the interface between psychiatry and neurology. PMID- 16251170 TI - Successful treatment of aspergillus brain abscess in a child with acute lymphoblastic leukemia and liver failure. AB - Invasive fungal infection continues to pose a significant threat to immunocompromised patients, with cerebral aspergillosis being among the most feared ones. The authors describe an adolescent girl with acute lymphoblastic leukemia (ALL) with subsequent acute liver failure, who developed an aspergillus brain abscess. The patient was treated with combined antifungal therapy using amphotericin B local instillation, prolonged systemic amphotericin B colloidal dispersion along with vinca alkaloids-containing chemotherapy, followed by neurosurgical debridement and oral voriconazole in the setting of ongoing antileukemic maintenance chemotherapy. Her ALL remains now in complete remission 30 months from diagnosis, with no evidence of fungal infection. PMID- 16251171 TI - The anemia of familial Mediterranean fever disease. AB - The aim of this study was to evaluate the incidence of anemia detected in familial Mediterranean fever (FMF) and the effect of disease activity and colchicine therapy along with interleukins to laboratory tests, including serum transferrin receptor (TfR), in the diagnosis of anemia seen in FMF in children. After detecting anemia in 63.4% of 172 FMF patients followed up by our rheumatology outpatient polyclinics, it was decided to study 3 groups of patients: group 1, 17 newly diagnosed FMF patients; group 2, 36 FMF patients on colchicine therapy; and group 3, 17 healthy children as control for the symptom of anemia. All 3 groups of patients were investigated for their hematological parameters, iron status, including soluble transferrin receptor (sTFR) concentrations and sTFR index, and IL-6 levels. Anemia ratio was 9/17, 53%; 11/36, 31%; and 1/17, 5% in the groups 1, 2 and 3, respectively. There was a significant difference between hemoglobin (Hb) values in the first group and the second (patients who were on colchicine therapy). Furthermore, in the second group there was a significant difference between the Hb concentrations at the time of diagnosis and after colchicine therapy (p = .003). There was a positive correlation between Hb and plasma iron and transferrin saturation in group 1 and disease beginning age, iron, transferrin saturation, and erythrocyte sedimentation rate (ESR) in the second group. In the first group the anemic patients' iron and transferrin saturation were significantly lower than normal, while ferritin levels were higher. In the second group, a good correlation was found with ESR and Hb levels; the higher ESR values were detected in patients with lower Hb values. Of the anemic and nonanemic patients of the first and second groups, values for interleukin 6 and iron parameters, including sTFR, were found similar. Anemia detected in FMF patients was found related to iron status more than interleukins. Colchicine therapy had a positive effect on anemia as well as on disease activity. Resolution of symptoms of FMF occurred with correction of the anemia, if the patient ESR values also decreased on colchicine therapy. PMID- 16251172 TI - Stepping up versus standard doses of erythropoietin in preterm infants: a randomized controlled trial. AB - In this study, it is hypothesized that a planned increase in the dose of recombinant human erythropoietin (rh-EPO) can prevent transfusion in very low birth weight infants. Two different regimens of rh-EPO were administrated, one consisting in increasing dosage up to 5000 U/kg/wk, according to the individual reticulocytes response, and the second in a standard therapy of 1250 U/kg/wk. Fifty-one infants participated. Despite a significant higher reticulocytosis, the study was prematurely terminated due to the results of an interim analysis showing that transfusion was not avoided by increasing the rh-EPO. No significant differences were found between the two regimens concerning transfusion rate, volume transfused, gain in weight, and adverse effects. Progressive titration of rh-EPO to improve the biological response does not leave premature infants free of transfusion. PMID- 16251174 TI - Discitis following lumbar puncture in non-Hodgkin lymphoma. AB - Discitis in children is a rare disorder of intervertebral disc and vertebral end plate. Infection or trauma, like lumbar puncture, may be the possible causes. Low back pain and gait disturbance are the main symptoms. The most appropriate diagnostic procedure is MRI. Treatment is mainly empirical. Here a case with non Hodgkin lymphoma is discussed. Treatment consisted of strict bed rest and antibiotics. Safe and sterile technique is important in patients with invasive procedures like intrathecal chemotherapy. Although discitis is a self-healing condition, it might cause vertebral osteomyelitis. In this regard, physicians should be aware of this probable complication after lumbar puncture and manage it earlier in children with cancer. PMID- 16251173 TI - Disseminated intravascular coagulation in pediatric patients: clinical and laboratory features and prognostic factors influencing the survival. AB - Although disseminated intravascular coagulation (DIC) has been a well-known disorder for many years, there is lack of sufficient number of clinical trials about incidence, frequency of underlying disorders, and prognosis of DIC in children. The aim of this study was to evaluate the frequency, etiologic factors, and clinical and laboratory findings of DIC and to determine the prognostic factors influencing the mortality in hospitalized pediatric patients. Medical records of 5535 children who were hospitalized were investigated. Sixty-two patients who were diagnosed as acute DIC were enrolled. The frequency of DIC was 1.12%. The underlying etiologic factors were infection in 59 patients (95.2%) and major trauma in 3 patients (4.8%). The frequency of bleeding and thrombosis was 48.8 and 4.8%. Respiratory, cardiovascular, hepatic, renal, neurologic, and gastrointestinal dysfunction was present in 71, 67.7, 35.5, 16.1, 16.1 and 11.3% of patients, respectively. Respiratory and cardiovascular dysfunctions were significantly associated with mortality. Multiorgan dysfunction syndrome (MODS) was present in 85.5% of the patients, and 54.8% of the patients had developed acute respiratory distress syndrome (ARDS). Mortality rate was significantly high in patients with MODS and ARDS. In multivariete logistic regression analysis, only ARDS and cardiovascular dysfunction had predictive and prognostic value on mortality. None of the diagnostic laboratory tests had predictive or prognostic value and the degree of abnormality of these tests did not show any correlation with mortality. In conclusion, DIC is not a rare disorder in hospitalized children, especially in patients with sepsis, and MODS, ARDS, and respiratory and cardiovascular system dysfunctions are poor prognostic factors. PMID- 16251175 TI - Successful treatment of relapsed multifocal nonvisceral infantile myofibromatosis. AB - Infantile myofibromatosis is a very rare tumor in childhood and infancy. The authors report on a 4-year-old boy who presented with two relapses of initially multifocal infantile myofibromatosis without visceral involvement. The lesions of the skull and the abdomen were excised while osteolytic lesions of the limbs were not treated. Chemotherapy or radiation have not been applicated. Three years after initial diagnosis there is no evidence for persistence or recurrence of the tumor. PMID- 16251176 TI - Poor outcome of two children with relapsed state stage IV alveolar rhabdomyosarcoma after allogeneic stem cell transplantation. AB - The authors report on 2 boys, 11(1/2) and 13 years old, who received allogeneic stem cell transplantation (alloSCT) from their HLA-identical sibling after relapse of stage IV alveolar rhabdomyosarcoma. Both patients were transplanted in a non-remission status. After alloSCT both patients experienced disease progression at the primary tumor location sites and died due to the underlying disease 146 and 379 days after transplantation. The authors conclude that an alloSCT derived graft versus tumor effect might not be effective enough to overcome alveolar rhabdomyosarcoma when transplantation is carried out in a nonremission status. PMID- 16251177 TI - Primary testicular Burkitt lymphoma in a child. AB - A 13-year-old boy was referred to the authors' hospital following a right inguinal orchiectomy for a right scrotal mass. Histopathological examination revealed Burkitt lymphoma. The left testis was found to be small with heterogeneous parenchyma by scrotal ultrasound (US) and other systemic investigations were negative for lymphoma involvement. Ultrasound-guided fine needle aspiration biopsy showed no evidence of involvement in the left testis. Considering stage I Burkitt lymphoma, chemotherapy was started. Following the first course, US findings changed: the volume of the left testis decreased and the parenchyma became homogeneous. The left testis was considered to be involved by lymphoma at initial diagnosis and chemotherapy was intensified. At the end of 5 months of chemotherapy the left testis was again heterogeneous in US. A wedge biopsy was negative for lymphoma. The patient is under regular follow-up and is in complete remission 19 months after the end of chemotherapy. Primary testicular lymphoma is quite rare in children and experience is limited. Changes in testicular size and parenchyma by US should not necessarily indicate involvement by lymphoma in pubertal boys. PMID- 16251178 TI - Serum soluble transferrin receptor is a valuable diagnostic tool in iron deficiency of breath-holding spells. AB - Iron-deficiency anemia may be a factor contributing to breath-holding spells. The serum transferrin receptor provides a useful measure of tissue iron deficiency. In this study of 50 breath-holders, while iron-deficiency anemia was detected in 28 (56%) of patients with routine tests, serum transferrin receptor levels were found increased in all patients. A positive correlation was detected between serum soluble transferrin receptor levels and frequency of attacks. It is suggested that the serum transferrin receptor level is useful as a single test for identification of iron deficiency in breath-holders. Moreover, if iron deficiency can be diagnosed earlier, then patients can be treated earlier. PMID- 16251179 TI - Cobalamin C disease presenting with hemophagocytic lymphohistiocytosis. AB - Cobalamin C disease is a rare genetic condition resulting in methylmalonic aciduria, homocystinuria, and hematologic abnormalities. Clinical characteristics include ophthalmologic findings and neurological abnormalities, such as microcephaly, seizure, and mental retardation. The authors report on a 4-month old patient initially diagnosed with hemophagocytic lymphohistiocytosis (HLH), who was later diagnosed with cobalamin C disease. PMID- 16251180 TI - Soft tissue density variations in thalassemia major: a possible pitfall in lumbar bone mineral density measurements by dual-energy X-ray absorptiometry. AB - Osteoporosis is common in patients with thalassemia major. A 16-year-old patient with thalassemia major was referred for evaluation of osteoporosis. The results of dual-energy X-ray absorptiometry in a patient with thalassemia major are presented. The patient underwent measurements of the spine in both AP and lateral position. The DXA scan of the spine in AP projection showed diffusely increased density overlying the lower thoracic and upper lumbar vertebrae. The overall density of L(2)-L(4) was 0.6465 g/cm(2) (4.93 SD as compared with young adults). The DXA scan of the spine in lateral projection showed diffusely increased density in front part of the spine. The BMD of the L(3) vertebra was 0.3669 g/cm(2) (0.30 SD as compared with young adults). It is important to interpret the images visually in order to obtain true values of BMD, and preclude invalid BMD measurements. PMID- 16251182 TI - Inhibition of HIV-1 maturation via drug association with the viral Gag protein in immature HIV-1 particles. AB - The small molecule 3-O-(3',3'-dimethylsuccinyl)-betulinic acid (DSB) potently inhibits human immunodeficiency virus, type 1 (HIV-1) replication by interfering with proteolytic cleavage of the viral Gag protein at a specific site. Here we have demonstrated that the antiviral mechanism involves the association of DSB with Gag at a 1:1 stoichiometry within immature HIV-1 particles. The binding was specific, as mutations in Gag that confer resistance to DSB inhibited the association, which could be competed by DSB but not by the inactive compound betulinic acid. The addition of DSB to purified immature viral cores inhibited the cleavage of Gag at the CA-SP1 junction in vitro, thus reproducing the effect of the drug when present during maturation of HIV-1 particles. Based on these findings, we propose a model in which a trimer of DSB associates with the CA-SP1 junction of adjacent subunits within the Gag polymer. The model may explain the ability of highly similar compounds to specifically target the seemingly unrelated steps of HIV-1 maturation and virus entry. PMID- 16251181 TI - Inhibition of hormone and cytokine-stimulated osteoclastogenesis and bone resorption by interleukin-4 and interleukin-13 is associated with increased osteoprotegerin and decreased RANKL and RANK in a STAT6-dependent pathway. AB - Interleukin (IL)-4 and IL-13 are cytokines that inhibit bone resorption. Data showing an inhibitory effect of IL-4 and IL-13 on RANK mRNA in mouse calvariae were first reported at the 22nd American Society for Bone and Mineral Research Meeting (Lerner, U.H., and Conaway, H. H. 2000) J. Bone Min. Res. 15, Suppl. 1, Abstr. SU 230). In the present study, release of 45Ca from cultured mouse calvarial bones stimulated by different cytokines, peptides, and steroid hormones was inhibited by IL-4 and IL-13. IL-4 and IL-13 decreased receptor activator of nuclear factor-kappaB ligand (RANKL) and RANK mRNA and increased osteoprotegerin (OPG) mRNA in calvariae. Additionally, the cytokines decreased RANKL protein and increased OPG protein in calvarial bones. In osteoblasts isolated from calvariae, both an increase in RANKL mRNA and a decrease in OPG mRNA and protein elicited by vitamin D3 were reversed by IL-4 and IL-13. IL-4 and IL-13 decreased the number of tartrate-resistant acid phosphatase positive multinucleated cells and the mRNA expression of calcitonin receptor, tartrate-resistant acid phosphatase, and cathepsin K in mouse spleen cells and bone marrow macrophages (BMM) treated with macrophage colony-stimulating factor and RANKL. Inhibition of mRNA for RANK and the transcription factor NFAT2 was also noted in spleen cell and BMM cultures treated with IL-4 and IL-13. In addition, RANK mRNA and RANK protein were decreased by IL-4 and IL-13 in RAW 264.7 cells. Osteoblasts, spleen cells, and BMM expressed mRNA for the four proteins making up the IL-4 and IL-13 receptors. No effects by IL-4 on bone resorption and osteoclast formation or on RANKL and RANK mRNA expression were seen in Stat6-/- mice. The data indicate that IL-4 and IL-13, via a STAT6-dependent pathway, inhibit osteoclast differentiation and bone resorption by activating receptors on osteoblasts and osteoclasts that affect the RANKL/RANK/OPG system. PMID- 16251183 TI - Galpha12/13 is essential for directed cell migration and localized Rho-Dia1 function. AB - Scratch-wound assays are frequently used to study directed cell migration, a process critical for embryogenesis, invasion, and tissue repair. The function and identity of trimeric G-proteins in cell behavior during wound healing is not known. Here we show that Galpha12/13, but not Galphaq/11 or Galphai, is indispensable for coordinated and directed cell migration. In mouse embryonic fibroblasts endogenous Rho activity is present at the rear of migrating cells but also at the leading edge, whereas it is undetectable at the cell front of Galpha12/13-deficient mouse embryonic fibroblasts. Spatial activation of Rho at the wound edge can be stimulated by lysophosphatidic acid. Active Rho colocalizes with the diaphanous-related formin Dia1 at the cell front. Galpha12/13-deficient cells lack Dia1 localization to the wound edge and are unable to form orientated, stable microtubules during wound healing. Knock down of Dia1 reveals its requirement for microtubule stabilization as well as polarized cell migration. Thus, we identified Galpha12/13-proteins as essential components linking extracellular signals to localized Rho-Dia1 function during directed cell movement. PMID- 16251184 TI - Wnt proteins prevent apoptosis of both uncommitted osteoblast progenitors and differentiated osteoblasts by beta-catenin-dependent and -independent signaling cascades involving Src/ERK and phosphatidylinositol 3-kinase/AKT. AB - Genetic studies in humans and mice have revealed an important role of the Wnt signaling pathway in the regulation of bone mass, resulting from potent effects on the control of osteoblast progenitor proliferation, commitment, differentiation, and perhaps osteoblast apoptosis. To establish the linkage between Wnts and osteoblast survival and to elucidate the molecular pathways that link the two, we have utilized three cell models: the uncommitted bipotential C2C12 cells, the pre-osteoblastic cell line MC3T3-E1, and bone marrow-derived OB 6 osteoblasts. Serum withdrawal-induced apoptosis was prevented by the canonical Wnts (Wnt3a and Wnt1) and the noncanonical Wnt5a in all cell types. Wnt3a induced LRP5-independent transient phosphorylation and nuclear accumulation of ERKs and phosphorylation of Src and Akt. The anti-apoptotic effect of Wnt3a was abrogated by inhibitors of canonical Wnt signaling, as well as by inhibitors of MEK, Src, phosphatidylinositol 3-kinase (PI3K), or Akt kinases, or by the addition of cycloheximide to the culture medium. Wnt3a-induced phosphorylation of GSK-3beta and downstream activation of beta-catenin-mediated transcription required ERK, PI3K, and Akt signaling. Wnt3a increased the expression of the anti-apoptotic protein Bcl-2 in an ERK-dependent manner. Beta-catenin-mediated transcription was permissive for the anti-apoptotic actions of Wnt1 and Wnt3a but was dispensable for the anti-apoptotic action of Wnt5a. However, Src, ERKs, PI3K, and Akt kinases were required for the anti-apoptotic effects of Wnt5a. These results demonstrate for the first time that Wnt proteins, irrespective of their ability to stimulate canonical Wnt signaling, prolong the survival of osteoblasts and uncommitted osteoblast progenitors via activation of the Src/ERK and PI3K/Akt signaling cascades. PMID- 16251185 TI - Kinetics and template nucleation of self-assembled hydroxyapatite nanocrystallites by chondroitin sulfate. AB - Biomineralization is an important process, which is often assisted by biomolecules. In this paper, the effect of chondroitin sulfate on the crystallization of hydroxyapatite was examined quantitatively based on a generic heterogeneous nucleation model. It is found that chondroitin sulfate can suppress the supersaturation-driven interfacial structure mismatch between the hydroxyapatite crystal and the substrate and promote the formation of ordered hydroxyapatite nanocrystallite assemblies. The nucleation mechanism of self aligned hydroxyapatite nanocrystallites was examined from the viewpoints of kinetics and interfacial structure and properties, which contributes to an understanding of the fundamentals of biomineralization of self-assembled structures. The results obtained from this study will provide a basic principle to design and fabricate highly orderly organic-inorganic hybrid materials. PMID- 16251186 TI - Toward a better understanding of the basis of the molecular mimicry of polysaccharide antigens by peptides: the example of Shigella flexneri 5a. AB - Protein conjugates of oligosaccharides or peptides that mimic complex bacterial polysaccharide antigens represent alternatives to the classical polysaccharide based conjugate vaccines developed so far. Hence, a better understanding of the molecular basis ensuring appropriate mimicry is required in order to design efficient carbohydrate mimic-based vaccines. This study focuses on the following two unrelated sets of mimics of the Shigella flexneri 5a O-specific polysaccharide (O-SP): (i) a synthetic branched pentasaccharide known to mimic the average solution conformation of S. flexneri 5a O-SP, and (ii) three nonapeptides selected upon screening of phage-displayed peptide libraries with two protective murine monoclonal antibodies (mAbs) of the A isotype specific for S. flexneri 5a O-SP. By inducing anti-O-SP antibodies upon immunization in mice when appropriately presented to the immune system, the pentasaccharide and peptides p100c and p115, but not peptide p22, were qualified as mimotopes of the native antigen. NMR studies based on transferred NOE (trNOE) experiments revealed that both kinds of mimotopes had an average conformation when bound to the mAbs that was close to that of their free form. Most interestingly, saturation transfer difference (STD) experiments showed that the characteristic turn conformations adopted by the major conformers of p100c and p115, as well as of p22, are clearly involved in mAb binding. These latter experiments also showed that the branched glucose residue of the pentasaccharide was a key part of the determinant recognized by the protective mAbs. Finally, by using NMR-derived pentasaccharide and peptide conformations coupled to STD information, models of antigen-antibody interaction were obtained. Most interestingly, only one model was found compatible with experimental data when large O-SP fragments were docked into one of the mIgA-binding sites. This newly made available system provides a new contribution to the understanding of the molecular mimicry of complex polysaccharides by peptides and short oligosaccharides. PMID- 16251187 TI - Identification of a lipid peroxidation product as a potential trigger of the p53 pathway. AB - The tumor suppressor and transcription factor p53 is a key modulator of cellular stress responses, and activation of p53 can trigger apoptosis in many cell types, including neurons. We found that this nuclear protein was significantly phosphorylated when human neuroblastoma SH-SY5Y cells were exposed to in vitro oxidized polyunsaturated fatty acids. To identify an oxidized lipid that induces p53 phosphorylation, we conducted a screening of lipid peroxidation products in human neuroblastoma SH-SY5Y cells and identified 4-oxo-2-nonenal (ONE), a recently identified aldehyde originating from the peroxidation of omega6 polyunsaturated fatty acids, as a potential inducer of the p53 phosphorylation. We also found that ONE induced the phosphorylation of ataxia telangiectasia mutated, which plays an essential role in transmitting DNA damage signals by the phosphorylation of p53. In addition, exposure of the cells to ONE resulted in an accumulation of ubiquitinated proteins and in a significant inhibition of proteasome activities, suggesting that ONE acted on the ubiquitin-proteasome pathway, a regulatory mechanism of p53 turnover. In addition, the observation that the ONE-induced p53 response was associated with the induction of apoptosis suggested that ONE activated the p53-dependent apoptosis mechanism via activation of the p53 signaling pathway and down-regulation of the p53 turnover. Finally, we observed that the ONE-2'-deoxyguanosine adduct, 7-(2-oxo-heptyl)-substituted 1,N(2)-etheno-2'-deoxyguanosine, was accumulated in the spinal cord motor neurons of patients with sporadic amyotrophic lateral sclerosis. These data may suggest the potential critical role for ONE in the induction of a neuronal apoptosis program during oxidative processes. PMID- 16251188 TI - Interleukin-8 secretion by fibroblasts induced by low density lipoproteins is p38 MAPK-dependent and leads to cell spreading and wound closure. AB - We have previously reported (Dobreva, I., Waeber, G., Mooser, V., James, R. W., and Widmann, C. (2003) J. Lipid Res. 44, 2382-2390) that low density lipoproteins (LDLs) induce activation of the p38 MAPK pathway, resulting in fibroblast spreading and lamellipodia formation. Here, we show that LDL-stimulated fibroblast spreading and wound sealing are due to secretion of a soluble factor. Using an antibody-based human protein array, interleukin-8 (IL-8) was identified as the main cytokine whose concentration was increased in supernatants from LDL stimulated cells. Incubation of supernatants from LDL-treated cells with an anti IL-8 blocking antibody completely abolished their ability to induce cell spreading and mediate wound closure. In addition, fibroblasts treated with recombinant IL-8 spread to the same extent as cells incubated with LDL or supernatants from LDL-treated cells. The ability of LDL and IL-8 to induce fibroblast spreading was mediated by the IL-8 receptor type II (CXCR-2). Furthermore, LDL-induced IL-8 production and subsequent wound closure required the activation of the p38 MAPK pathway, because both processes were abrogated by a specific p38 inhibitor. Therefore, the capacity of LDLs to induce fibroblast spreading and accelerate wound closure relies on their ability to stimulate IL-8 secretion in a p38 MAPK-dependent manner. Regulation of fibroblast shape and migration by lipoproteins may be relevant to atherosclerosis that is characterized by increased LDL cholesterol levels, IL-8 production, and extensive remodeling of the vessel wall. PMID- 16251190 TI - Regulation of the INK4a/ARF locus by histone deacetylase inhibitors. AB - Despite the importance of the INK4a/ARF locus in tumor suppression, its modulation by histone deacetylase inhibitors (HDACis) remains to be characterized. Here, we have shown that the levels of p16INK4a are decreased in human and murine fibroblasts upon exposure to relatively high concentrations of trichostatin A and sodium butyrate. Interestingly, the levels of p19ARF are strongly upregulated in murine cells even at low concentrations of HDACis. Using ARF-deficient cells, we have demonstrated that p19ARF plays an active role in HDACi-triggered cytostasis and the contribution of p19ARF to this arrest is of higher magnitude than that of the well established HDACi target p21Waf1/Cip. Moreover, chemically induced fibrosarcomas in ARF-null mice are more resistant to the therapeutic effect of HDACis than similar tumors in wild type or p21Waf1/Cip null mice. Together, our results have established the tumor suppressor ARF as a relevant target for HDACi chemotherapy. PMID- 16251189 TI - GPX2, encoding a phospholipid hydroperoxide glutathione peroxidase homologue, codes for an atypical 2-Cys peroxiredoxin in Saccharomyces cerevisiae. AB - We have previously reported that Saccharomyces cerevisiae has three glutathione peroxidase homologues (GPX1, GPX2, and GPX3) (Inoue, Y., Matsuda, T., Sugiyama, K., Izawa, S., and Kimura, A. (1999) J. Biol. Chem. 274, 27002-27009). Of these, the GPX2 gene product (Gpx2) shows the greatest similarity to phospholipid hydroperoxide glutathione peroxidase. Here we show that GPX2 encodes an atypical 2-Cys peroxiredoxin which uses thioredoxin as an electron donor. Gpx2 was essentially in a reduced form even in mutants defective in glutathione reductase or glutaredoxin under oxidative stressed conditions. On the other hand, Gpx2 was partially oxidized in a mutant defective in cytosolic thioredoxin (trx1Deltatrx2Delta) under non-stressed conditions and completely oxidized in tert-butyl hydroperoxide-treated cells of trx1Deltatrx2Delta and thioredoxin reductase-deficient mutant cells. Alanine scanning of cysteine residues of Gpx2 revealed that an intramolecular disulfide bond was formed between Cys37 and Cys83 in vivo. Gpx2 was purified to determine whether it functions as a peroxidase that uses thioredoxin as an electron donor in vitro. Gpx2 reduced H2O2 and tert-butyl hydroperoxide in the presence of thioredoxin, thioredoxin reductase, and NADPH (for H2O2, Km= 20 microm, kcat = 9.57 x 10(2) s(-1); for tert-butyl hydroperoxide, Km= 62.5 microm, kcat = 3.68 x 10(2) s(-1)); however, it showed remarkably less activity toward these peroxides in the presence of glutathione, glutathione reductase, and NADPH. The sensitivity of yeast cells to tert-butyl hydroperoxide was found to be exacerbated by the co-existence of Ca2+, a tendency that was most obvious in gpx2Delta cells. Although the redox state of Gpx2 was not affected by Ca2+, the Gpx2 level was markedly increased in the presence of both tert-butyl hydroperoxide and Ca2+. Gpx2 is likely to play an important role in the protection of cells from oxidative stress in the presence of Ca2+. PMID- 16251191 TI - Characterization of a binary tandem domain F-type lectin from striped bass (Morone saxatilis). AB - Among other functions, lectins play an important role in the innate immune response of vertebrates and invertebrates by recognizing exposed glycans on the surface of potential pathogens. Despite the typically weak interaction of lectin domains with their carbohydrate ligands, they usually achieve high avidity through oligomeric structures or by the presence of tandem carbohydrate-binding domains along the polypeptide. The recently described structure of the fucose binding European eel agglutinin revealed a novel lectin fold (the "F-type" fold), which is shared with other carbohydrate-binding proteins and apparently unrelated proteins from prokaryotes to vertebrates, and a unique fucose-binding sequence motif. Here we described the biochemical and molecular characterization of a unique fucose-binding lectin (MsaFBP32) isolated from serum of the striped bass (Morone saxatilis), composed of two tandem domains that exhibit the eel carbohydrate recognition sequence motif, which we designate F-type. We also described a novel lectin family ("F-type") constituted by a large number of proteins exhibiting greater multiples of the F-type motif, either tandemly arrayed or in mosaic combinations with other domains, including a putative transmembrane receptor, that suggests an extensive functional diversification of this lectin family. Among the tandem lectins, MsaFBP32 and other tandem binary homologues appear unique in that although their N-terminal domain shows close similarity to the fucose recognition domain of the eel agglutinin, their C terminal domain exhibits changes that potentially could confer a distinct specificity for fucosylated ligands. In contrast with the amniotes, in which the F-type lectins appear conspicuously absent, the widespread gene duplication in the teleost fish suggests these F-type lectins acquired increasing evolutionary value within this taxon. PMID- 16251192 TI - Regulation of transforming growth factor-beta signaling and PDK1 kinase activity by physical interaction between PDK1 and serine-threonine kinase receptor associated protein. AB - To gain more insights about the biological roles of PDK1, we have used the yeast two-hybrid system and in vivo binding assay to identify interacting molecules that associate with PDK1. As a result, serine-threonine kinase receptor associated protein (STRAP), a transforming growth factor-beta (TGF-beta) receptor interacting protein, was identified as an interacting partner of PDK1. STRAP was found to form in vivo complexes with PDK1 in intact cells. Mapping analysis revealed that this binding was only mediated by the catalytic domain of PDK1 and not by the pleckstrin homology domain. Insulin enhanced a physical association between PDK1 and STRAP in intact cells, but this insulin-induced association was prevented by wortmannin, a phosphatidylinositol 3-kinase inhibitor. In addition, the association between PDK1 and STRAP was decreased by TGF-beta treatment. Analysis of the activities of the interacting proteins showed that PDK1 kinase activity was significantly increased by coexpression of STRAP, probably through the inhibition of the binding of 14-3-3, a negative regulator, to PDK1. Consistently, knockdown of the endogenous STRAP by the transfection of the small interfering RNA resulted in the decrease of PDK1 kinase activity. PDK1 also exhibited an inhibition of TGF-beta signaling with STRAP by contributing to the stable association between TGF-beta receptor and Smad7. Moreover, confocal microscopic study and immunostaining results demonstrated that PDK1 prevented the nuclear translocation of Smad3 in response to TGF-beta. Knockdown of endogenous PDK1 with small interfering RNA has an opposite effect. Taken together, these results suggested that STRAP acts as an intermediate signaling molecule linking between the phosphatidylinositol 3-kinase/PDK1 and the TGF-beta signaling pathways. PMID- 16251193 TI - Centrosomal pericentrin is a direct cleavage target of membrane type-1 matrix metalloproteinase in humans but not in mice: potential implications for tumorigenesis. AB - Membrane type-1 matrix metalloproteinase (MT1-MMP) exhibits distinctive and important pericellular cleavage functions. Recently, we determined that MT1-MMP was trafficked to the centrosomes in the course of endocytosis. Our data suggested that the functionally important, integral, centrosomal protein, pericentrin-2, was a cleavage target of MT1-MMP in human and in canine cells and that the sequence of the cleavage sites were ALRRLLG1156 downward arrow L1157FG and ALRRLLS2068 downward arrow L2069FG, respectively. The presence of Asp-948 at the P1 position inactivated the corresponding site (ALRRLLD948-L949FGD) in murine pericentrin. To confirm that MT1-MMP itself cleaves pericentrin directly, rather than indirectly, we analyzed the cleavage of the peptides that span the MT1-MMP cleavage site. In addition, we analyzed glioma U251 cells, which co-expressed MT1 MMP with the wild type murine pericentrin and the D948G mutant. We determined that the D948G mutant that exhibited the cleavage sequence of human pericentrin was sensitive to MT1-MMP, whereas unmodified murine pericentrin was resistant to proteolysis. Taken together, our results confirm that MT1-MMP cleaves pericentrin 2 in humans but not in mice and that mouse models of cancer probably cannot be used to critically examine MT1-MMP functionality. PMID- 16251194 TI - ORF17 from the clavulanic acid biosynthesis gene cluster catalyzes the ATP dependent formation of N-glycyl-clavaminic acid. AB - (3R,5R)-Clavulanic acid, a clinically used inhibitor of serine beta-lactamases, is produced by fermentation of Streptomyces clavuligerus. The early steps in clavulanic acid biosynthesis leading to the bicyclic beta-lactam intermediate (3S,5S)-clavaminic acid have been defined. However, the mechanism by which (3S,5S)-clavaminic acid is converted to the penultimate intermediate (3R,5R) clavaldehyde is unclear. Disruption of orf15 or orf16, of the clavulanic acid biosynthesis gene cluster, blocks clavulanic acid production and leads to the accumulation of N-acetyl-glycyl-clavaminic acid and N-glycyl-clavaminic acid, suggesting that these compounds are intermediates in the pathway. Two alternative start codons have been proposed for orf17 to encode for two possible polypeptides, one of which has 92 N-terminal residues less then the other. The shorter version of orf17 was successfully expressed in Escherichia coli and purified as a monomeric protein. Sequence analyses predicting the ORF17 protein to be a member of the ATP-grasp fold superfamily were supported by soft ionization mass spectrometric analyses that demonstrated binding of ATP to the ORF17 protein. Semisynthetic clavaminic acid, prepared by in vitro reconstitution of the biosynthetic pathway from the synthetically accessible intermediate proclavaminic acid, was shown by mass spectrometric analyses to be converted to N glycyl-clavaminic acid in the presence of ORF17, ATP, and glycine. Under the same conditions N-acetyl-glycine and clavaminic acid were not converted to N-acetyl glycyl-clavaminic acid. The specificity of ORF17 as an N-glycyl-clavaminic acid synthetase, together with the reported accumulation of N-glycyl-clavaminic acid in orf15 and orf16 disruption mutants, suggested that N-glycyl-clavaminic acid is an intermediate in clavulanic acid biosynthesis. PMID- 16251196 TI - Defective lysosomal arginine transport in juvenile Batten disease. AB - Mutations in the CLN3 gene, which encodes a lysosomal membrane protein, are responsible for the neurodegenerative disorder juvenile Batten disease. A previous study on the yeast homolog to CLN3, designated Btn1p, revealed a potential role for CLN3 in the transport of arginine into the yeast vacuole, the equivalent organelle to the mammalian lysosome. Lysosomes isolated from lymphoblast cell lines, established from individuals with juvenile Batten disease bearing mutations in CLN3, but not age-matched controls, demonstrate defective transport of arginine. Furthermore, we show that there is a depletion of arginine in cells derived from individuals with juvenile Batten disease. We have, therefore, characterized lysosomal arginine transport in normal lysosomes and show that it is ATP-, v-ATPase- and cationic-dependent. This and previous studies have shown that both arginine and lysine are transported by the same transport system, designated system c. However, we report that lysosomes isolated from juvenile Batten disease lymphoblasts are only defective for arginine transport. These results suggest that the CLN3 defect in juvenile Batten disease may affect how intracellular levels of arginine are regulated or distributed throughout the cell. This assertion is supported by two other experimental approaches. First, an antibody to CLN3 can block lysosomal arginine transport and second, expression of CLN3 in JNCL cells using a lentiviral vector can restore lysosomal arginine transport. CLN3 may have a role in regulating intracellular levels of arginine possibly through control of the transport of this amino acid into lysosomes. PMID- 16251195 TI - The Pro-regions of lysyl oxidase and lysyl oxidase-like 1 are required for deposition onto elastic fibers. AB - These studies were undertaken to determine how lysyl oxidase (LOX) and lysyl oxidase like-1 (LOXL) enzymes are targeted to their substrates in the extracellular matrix. Full-length LOX/LOXL and constructs containing just the pro regions of each enzyme localized to elastic fibers when expressed in cultured cells. However, the LOXL catalytic domain without the pro-region was secreted into the medium but did not associate with matrix. Ligand blot and mammalian two hybrid assays confirmed an interaction between tropoelastin and the pro-regions of both LOX and LOXL. Immunofluorescence studies localized both enzymes to elastin at the earliest stages of elastic fiber assembly. Our results showed that the pro-regions of LOX and LOXL play a significant role in directing the deposition of both enzymes onto elastic fibers by mediating interactions with tropoelastin. These findings confirmed that an important element of substrate recognition lies in the pro-domain region of the molecule and that the pro-form of the enzyme is what initially interacts with the matrix substrate. These results have raised the interesting possibility that sequence differences between the pro-domain of LOX and LOXL account for some of the functional differences observed for the two enzymes. PMID- 16251197 TI - TAB2, TRAF6 and TAK1 are involved in NF-kappaB activation induced by the TNF receptor, Edar and its adaptator Edaradd. AB - Activation of the NF-kappaB pathway by the TNF-receptor Edar (Ectodysplasin receptor) and its downstream adaptator Edaradd (Edar-associated death domain) is essential for the development of hair follicles, teeth, exocrine glands and other ectodermal derivatives. Dysfunction of Edar signalling causes hypohidrotic/anhidrotic ectodermal dysplasia (ED), a disorder characterized by sparse hair, lack of sweat glands and malformation of teeth. The Edar signalling pathway stimulates NF-kappaB transcription factors via an activation of the IkappaB kinase (IKK) complex. To gain further insight into the mechanism of IKK activation by Edar and Edaradd, we performed a yeast two-hybrid screen and isolated TAB2 (TAK1-binding protein 2) as a binding partner of Edaradd. TAB2 is an adaptator protein that brigdes TRAF6 (TNF-receptor-associated factor 6) to TAK1 (TGFbeta-activated kinase 1), allowing TAK1 activation and subsequent IKK activation. Here, we show that endogenous and overexpressed TAB2, TRAF6 and TAK1 co-immunoprecipitated with Edaradd in 293 cells. Moreover, we show that dominant negative forms of TAB2, TRAF6 and TAK1 blocked the NF-kappaB activation induced by Edaradd. These results support the involvement of the TAB2/TRAF6/TAK1 signalling complex in the Edar signal transduction pathway and have important implications for our understanding of NF-kappaB activation and EDs in human. PMID- 16251198 TI - Gene expression variation and expression quantitative trait mapping of human chromosome 21 genes. AB - Inter-individual differences in gene expression are likely to account for an important fraction of phenotypic differences, including susceptibility to common disorders. Recent studies have shown extensive variation in gene expression levels in humans and other organisms, and that a fraction of this variation is under genetic control. We investigated the patterns of gene expression variation in a 25 Mb region of human chromosome 21, which has been associated with many Down syndrome (DS) phenotypes. Taqman real-time PCR was used to measure expression variation of 41 genes in lymphoblastoid cells of 40 unrelated individuals. For 25 genes found to be differentially expressed, additional analysis was performed in 10 CEPH families to determine heritabilities and map loci harboring regulatory variation. Seventy-six percent of the differentially expressed genes had significant heritabilities, and genomewide linkage analysis led to the identification of significant eQTLs for nine genes. Most eQTLs were in trans, with the best result (P=7.46 x 10(-8)) obtained for TMEM1 on chromosome 12q24.33. A cis-eQTL identified for CCT8 was validated by performing an association study in 60 individuals from the HapMap project. SNP rs965951 located within CCT8 was found to be significantly associated with its expression levels (P=2.5 x 10(-5)) confirming cis-regulatory variation. The results of our study provide a representative view of expression variation of chromosome 21 genes, identify loci involved in their regulation and suggest that genes, for which expression differences are significantly larger than 1.5-fold in control samples, are unlikely to be involved in DS-phenotypes present in all affected individuals. PMID- 16251199 TI - A segment of the Mecp2 promoter is sufficient to drive expression in neurons. AB - Rett syndrome (RTT) is caused by mutations in the gene encoding methyl CpG binding protein 2 (MeCP2). Although MeCP2 shows widespread expression in both neuronal and non-neuronal tissues, the symptoms of RTT are largely neurological. Herein, we have identified the regulatory region of the mouse Mecp2 gene that is sufficient for its restricted expression in neurons. A segment of the Mecp2 gene (-677/+56) exhibited strong promoter activity in neuronal cell lines and cortical neurons, but was inactive in non-neuronal cells and glia. The region necessary for neuronal-specific promoter activity was located within a 19 bp region (-63/ 45). Several nuclear factors were found to bind to this region and some of these factors were enriched in nuclear extracts prepared from the brain. To examine the activity of the Mecp2 promoter in vivo, we generated transgenic mice expressing the LacZ reporter driven by the -677/+56 region of the Mecp2 gene. The transgene was expressed in the mesencephalon as early as embryonic day 10 and in the hindbrain and spinal cord by E12. Interestingly, a marked induction of transgene expression was observed postnatally throughout the brain, similar to that of endogenous MeCP2. However, expression of the transgene was absent in non-neuronal tissues that are known to express Mecp2. Taken together, these data indicate that the -677/+56 region of the Mecp2 promoter partially recapitulates the native expression pattern of the Mecp2 gene, which possesses restricted expression in neurons of the central nervous system. PMID- 16251200 TI - Fulvestrant, a new treatment option for advanced breast cancer: tolerability versus existing agents. AB - Owing to its favourable tolerability profile versus cytotoxic chemotherapy, endocrine therapy is the treatment of choice for postmenopausal women with hormone receptor-positive advanced breast cancer (ABC). However, tolerability concerns associated with some endocrine treatments and the potential for cross resistance has helped to drive the need for new, effective and better-tolerated agents. Fulvestrant is a new type of oestrogen receptor antagonist with no agonist effects. In phase III trials, fulvestrant has been shown to be at least as effective as the third-generation aromatase inhibitor (AI) anastrozole in the treatment of postmenopausal women with ABC progressing on prior tamoxifen therapy. Fulvestrant is administered as a once-monthly 250 mg intramuscular injection into the gluteus muscle. Here we review the tolerability of fulvestrant in the treatment of postmenopausal women with hormone-sensitive ABC and compare it with that of the four most frequently prescribed endocrine treatments for advanced disease (tamoxifen, anastrozole, letrozole and exemestane). Compared with these agents, fulvestrant is well tolerated and is associated with a lower incidence of joint disorders compared with the non-steroidal AIs and none of the potential androgenic side-effects that are sometimes seen with steroidal AIs. It is also associated with hot flushes compared with tamoxifen. Fulvestrant therefore provides clinicians and patients with a useful, well-tolerated option for the treatment of hormone-sensitive ABC. Integration of such agents into the endocrine treatment sequence may extend the opportunity for using well-tolerated therapies before chemotherapy needs to be considered and thus may improve quality of life for patients with ABC. The overall safety profiles of newer agents such as fulvestrant will become increasingly clear with their ongoing use. PMID- 16251201 TI - Phase II study of tailored chemotherapy for advanced colorectal cancer with either 5-fluouracil and leucovorin or oxaliplatin and irinotecan based on the expression of thymidylate synthase and dihydropyrimidine dehydrogenase. AB - BACKGROUND: Thymidylate synthase (TS) and dihydropyrimidine dehydrogenase (DPD) are essential enzymes for 5-fluorouracil (5-FU) metabolism. In patients with advanced colorectal cancer (ACRC), retrospective studies have shown that low expression levels of TS and DPD correlated with response to 5-FU. We performed a prospective study in which the choice of first-line chemotherapy with either 5-FU or a non-5-FU containing regimen was based on TS and DPD expression. PATIENTS AND METHODS: Fresh-frozen samples of metastases were obtained from 58 previously untreated patients with ACRC. mRNA expression of TS and DPD was quantified using an RT-PCR assay. Patients with low tumor expression of both TS and DPD received weekly bolus 5-FU/leucovorin (LV) 500 mg/m2 (group A); patients with high TS and/or DPD received 3-weekly oxaliplatin 85 mg/m2 and irinotecan 200 mg/m2 (group B). After progression, cross-over to the alternative regimen was attempted. RESULTS: Of 53 eligible patients, 31 had tumors with both low TS and low DPD, and were treated in group A. A response was observed in 11 patients [35%; 95% confidence interval (CI) 19% to 54%]. Cross-over to second-line oxaliplatin/irinotecan resulted in a partial response in two out of 16 patients (13%; 95% CI 1% to 38%). In group B, four out of 22 patients responded (18%; 95% CI 5% to 40%), while no responses were observed in 12 patients after cross-over to 5-FU/LV (0%; 95% CI 0% to 28%). CONCLUSIONS: Prospective selection of 5-FU/LV chemotherapy based on low TS and DPD expression in patients with ACRC did not confirm the high response rates reported in retrospective studies. The procedure of obtaining metastatic tissue and quantitation of enzymes appeared feasible but cumbersome. Before assessing the clinical utility of a predictive marker in a randomized trial, future studies should focus on prospective validation of the assay in a large and well defined population. PMID- 16251202 TI - Correlation between anemia and functional/cognitive capacity in elderly lung cancer patients treated with chemotherapy. AB - BACKGROUND: Elderly cancer patients are often excluded from clinical trials and no data are available on the impact of chemotherapy-related anemia on their functional status and cognitive functions. This observational study investigates the association between hemoglobin (Hb) level and comprehensive geriatric assessment (CGA) variables (MMSE, ADL/IADL, GDS, CIRS and VAS). PATIENTS AND METHODS: We enrolled 42 consecutive lung cancer elderly patients undergoing chemotherapy that were evaluated at baseline and after each CT cycle at least until cycle 2. Hb association with CGA indexes was expressed using Spearman's non parametric coefficient r. RESULTS: Higher Hb values were significantly associated with more favourable values of all indexes measuring mental and functional capacity, depression and comorbidities. For all indexes except IADL, improvements from baseline were significantly related with concomitant Hb increases. In 14 patients given erythropoietin during the first two cycles, mean Hb increased from 9.2 to 10.8 g/dl, and the mean values of all CGA indexes were improved. On the contrary, in 18 patients not given erythropoietin, Hb varied from 13.0 to 11.2 g/dl and a parallel worsening in all CGA indexes was observed. CONCLUSIONS: Chemotherapy-related anemia is associated with impairment of functional status and cognitive functions. In elderly cancer patients anemia correction or maintenance could be useful to preserve functional independency and protect from mental decay. However, the study results need to be confirmed on a larger series of patients within a controlled clinical trial. PMID- 16251203 TI - Diverging trends in breast cancer mortality within Switzerland. AB - BACKGROUND: Substantial regional heterogeneity in the provision of cancer prevention and management services results from the decentralised Swiss healthcare system. MATERIALS AND METHODS: Breast cancer mortality trends between 1980 and 2002 were compared in two French- and in two German-speaking female populations of Switzerland, aged 55-74 years, characterised by different access to, and use of, mammography screening. RESULTS: Since the early 1990s, a 30% fall was observed in the French-speaking regions of Vaud and Geneva, where mammography screening is widespread, with no decline in the German-speaking areas of Basel and Zurich. CONCLUSION: Modification in breast cancer diagnosis and management in selected regions of Switzerland is urgently needed. PMID- 16251204 TI - Long-term cardiac toxicity after adjuvant epirubicin-based chemotherapy in early breast cancer: French Adjuvant Study Group results. AB - BACKGROUND: The aim of the study was to evaluate and compare incidence and risk factors of left ventricular dysfunction (LVD) in early breast cancer patients receiving (E+) or not (E-) epirubicin-based adjuvant chemotherapy. PATIENTS AND METHODS: Among eight FASG trials, 3577 assessable patients were analyzed retrospectively: 2553 received epirubicin, 662 received hormonotherapy alone and 362 had no systemic treatment. Chemotherapy was FEC regimen in 86% of cases (fluorouracil, epirubicin, cyclophosphamide). Epirubicin cumulative dose was < 300 mg/m2 in 1040 patients, 300-600 in 1155, > or = 600 in 279, followed by radiotherapy in 96% of cases. RESULTS: Twenty delayed LVD occurred: two in E- patients and 18 in E+ patients. In E+ patients, 14 patients normalized their cardiac function or did not require further investigations, one patient was stabilized with specific treatment, two patients worsened their functions and one died of congestive heart failure. The 7-year risk of LVD was 1.36% (95% CI 0.85 1.87) in E+ patients and 0.21% (95%CI: 0.00-0.52) in E- patients (P = 0.004). Two significant risk factors were identified: age > or = 65 years and body mass index > 27 kg/m2. CONCLUSION: After a long-term follow-up, epirubicin-related LVD risk was acceptable (1.36%) with one toxic death (0.04%). In 78% of cases, LVD were transient or well controlled. PMID- 16251205 TI - Catheter-associated thrombosis: thromboprophylaxis for cancer patients who carry factor V Leiden? PMID- 16251206 TI - Sequence specificity of Cr(III)-DNA adduct formation in the p53 gene: NGG sequences are preferential adduct-forming sites. AB - Hexavalent chromium [Cr(VI)] is a known etiological factor in human lung cancer. Cr(VI) exposure-related lung cancer has a high mutation incidence in the p53 gene. Upon intake in human cells Cr(VI) is reduced to Cr(III), which is able to conjugate with amino acids and consequently form either binary Cr(III)-DNA or ternary Cr(III)-amino acid-DNA adducts. Both binary and ternary Cr(III)-DNA adducts are mutagenic. We have found that the Escherichia coli nucleotide excision enzyme UvrABC nuclease is able to incise Cr(III)- and Cr(III)-histidine modified plasmid DNA and the extent of incision is proportional to the amount of Cr(III)-DNA adducts in the plasmid. In order to determine the role of Cr(III)-DNA adducts in the mutagenesis of the p53 gene in human cancer using the UvrABC nuclease incision method, we have mapped the Cr(III)-DNA distribution in PCR DNA fragments amplified from exons 5, 7 and 8 of the p53 gene. We have found that the sequence specificities of Cr(III)-DNA and Cr(III)-histidine-DNA adducts in the p53 gene sequence are identical and that both types of adducts are preferentially formed at -NGG- sequences, including codons 245, 248 and 249, the mutational hotspots in human lung cancer. It has been found that Cr(III)-DNA adducts induce mainly G to T mutations. Therefore, these results suggest that Cr(III)-DNA adduct formation contributes to the p53 gene mutations in lung carcinogenesis. PMID- 16251207 TI - Association of the CASP10 V410I variant with reduced familial breast cancer risk and interaction with the CASP8 D302H variant. AB - Dysregulation of apoptosis plays a crucial role in carcinogenesis. As part of death receptor- and mitochondrion-mediated apoptosis, the homologues caspases 10 and 8 may act as low-penetrance breast cancer (BC) susceptibility genes. In death receptor-mediated apoptosis, engagement of death receptors by their ligands involves the assembly of the death-inducing signalling complex (DISC). In mitochondrion-mediated apoptosis, the release of cytochrome c into the cytosol results in apoptosome formation. Recruitment of both caspases 10 and 8 (CASP10 and CASP8, respectively) to DISC and apoptosome leads to their activation by dimerization. We investigated the influence of the coding CASP10 variant V410I (G1228A) by performing a case-control study - using 511 familial BC cases and 547 control subjects - on BC risk and revealed a significant association of V410I with a reduced risk (OR = 0.62, 95% CI = 0.43-0.88, P = 0.0076) related to the number of variant alleles (P(trend) = 0.0039). As CASP10 and CASP8 functionally co-operate during apoptosis, we analysed the mutual effect of both CASP10 V410I and CASP8 D302H, resulting in a significant association between the number of the variant alleles I410 and H302 and a highly decreased familial BC risk (OR = 0.35, P(trend) = 0.007), pointing to the interaction between the CASP10 and CASP8 polymorphisms in breast carcinogenesis. PMID- 16251208 TI - SRC-1, a non-receptor type of protein tyrosine kinase, controls the direction of cell and growth cone migration in C. elegans. AB - Src family tyrosine kinase (SFK) has been implicated in the regulation of cell adhesion and migration during animal development. We show that SRC-1, an ortholog of SFK, plays an essential role in directing cell migration in Caenorhabditis elegans. The mutation in the src-1 gene results in defective distal tip cell (DTC)-directed gonad morphogenesis in an activity-dependent and DTC cell autonomous manners. In the src-1 mutants, DTCs fail to turn and continue their centrifugal migration along the ventral muscles. The effect of the src-1 mutation is suppressed by mutations in genes that function in the CED/Rac pathway, suggesting that SRC-1 in DTCs is an upstream regulator of a Rac pathway that controls cytoskeletal remodeling. In the src-1 mutant, the expression of unc 5/netrin receptor is normally regulated, and neither the precocious expression of UNC-5 nor the mutation in the unc-5 gene significantly affects the DTC migration defect. These data suggest that SRC-1 acts in the netrin signaling in DTCs. The src-1 mutant also exhibits cell-autonomous defects in the migration and growth cone path-finding of Q neuroblast descendants AVM and PVM. However, these roles of SRC-1 do not appear to involve the CED/Rac pathway. These findings show that SRC-1 functions in responding to various extracellular guidance cues that direct the cell migration via disparate signaling pathways in different cell types. PMID- 16251209 TI - Fgf signaling instructs position-dependent growth rate during zebrafish fin regeneration. AB - During appendage regeneration in urodeles and teleosts, tissue replacement is precisely regulated such that only the appropriate structures are recovered, a phenomenon referred to as positional memory. It is believed that there exists, or is quickly established after amputation, a dynamic gradient of positional information along the proximodistal (PD) axis of the appendage that assigns region-specific instructions to injured tissue. These instructions specify the amount of tissue to regenerate, as well as the rate at which regenerative growth is to occur. A striking theme among many species is that the rate of regeneration is more rapid in proximally amputated appendages compared with distal amputations. However, the underlying molecular regulation is unclear. Here, we identify position-dependent differences in the rate of growth during zebrafish caudal fin regeneration. These growth rates correlate with position-dependent differences in blastemal length, mitotic index and expression of the Fgf target genes mkp3, sef and spry4. To address whether PD differences in amounts of Fgf signaling are responsible for position-dependent blastemal function, we have generated transgenic fish in which Fgf receptor activity can be experimentally manipulated. We find that the level of Fgf signaling exhibits strict control over target gene expression, blastemal proliferation and regenerative growth rate. Our results demonstrate that Fgf signaling defines position-dependent blastemal properties and growth rates for the regenerating zebrafish appendage. PMID- 16251210 TI - Retinoic acid regulates the expression of dorsoventral topographic guidance molecules in the chick retina. AB - Asymmetric expression of several genes in the early eye anlagen is required for the dorsoventral (DV) and anteroposterior (AP) patterning of the retina. Some of these early patterning genes play a role in determining the graded expression of molecules that are needed to form the retinotectal map. The polarized expression of retinoic acid synthesizing and degrading enzymes along the DV axis in the retina leads to several zones of varied retinoic acid (RA) activity. This is suggestive of RA playing a role in DV patterning of the retina. A dominant negative form of the retinoic acid receptor alpha (DNhRARalpha) was expressed in the chick retina to block RA activity. RA signaling was found to play a role in regulating the expression of EphB2, EphB3 and ephrin B2, three molecules whose graded expression in the retina along the DV axis is important for establishing the correct retinotectal map. Blocking RA signaling by misexpression of a RA degrading enzyme, Cyp26A1 recapitulated some but not all the effects of DNhRARalpha. It also was found that Vax, a ventrally expressed transcription factor that regulates the expression of the EphB and ephrin B molecules, functions upstream of, or in parallel to, RA. Expression of DNhRARalpha led to increased levels of RA-synthesizing enzymes and loss of RA-degrading enzymes. Activation of such compensatory mechanisms when RA activity is blocked suggests that RA homeostasis is very strictly regulated in the retina. PMID- 16251211 TI - Homeotic factor ATBF1 induces the cell cycle arrest associated with neuronal differentiation. AB - The present study aimed to elucidate the function of AT motif-binding factor 1 (ATBF1) during neurogenesis in the developing brain and in primary cultures of neuroepithelial cells and cell lines (Neuro 2A and P19 cells). Here, we show that ATBF1 is expressed in the differentiating field in association with the neuronal differentiation markers beta-tubulin and MAP2 in the day E14.5 embryo rat brain, suggesting that it promotes neuronal differentiation. In support of this, we show that ATBF1 suppresses nestin expression, a neural stem cell marker, and activates the promoter of Neurod1 gene, a marker for neuronal differentiation. Furthermore, we show that in Neuro 2A cells, overexpressed ATBF1 localizes predominantly in the nucleus and causes cell cycle arrest. In P19 cells, which formed embryonic bodies in the floating condition, ATBF1 is mainly cytoplasmic and has no effect on the cell cycle. However, the cell cycle was arrested when ATBF1 became nuclear after transfer of P19 cells onto adhesive surfaces or in isolated single cells. The nuclear localization of ATBF1 was suppressed by treatment with caffeine, an inhibitor of PI(3)K-related kinase activity of ataxa-telangiectasia mutated (ATM) gene product. The cytoplasmic localization of ATBF1 in floating/nonadherent cells is due to CRM1-dependent nuclear export of ATBF1. Moreover, in the embryonic brain ATBF1 was expressed in the cytoplasm of proliferating stem cells on the ventricular zone, where cells are present at high density and interact through cell-to-cell contact. Conversely, in the differentiating field, where cell density is low and extracellular matrix is dense, the cell-to-matrix interaction triggered nuclear localization of ATBF1, resulting in the cell cycle arrest. We propose that ATBF1 plays an important role in the nucleus by organizing the neuronal differentiation associated with the cell cycle arrest. PMID- 16251212 TI - Cellular and molecular analyses of vascular tube and lumen formation in zebrafish. AB - Tube and lumen formation are essential steps in forming a functional vasculature. Despite their significance, our understanding of these processes remains limited, especially at the cellular and molecular levels. In this study, we analyze mechanisms of angioblast coalescence in the zebrafish embryonic midline and subsequent vascular tube formation. To facilitate these studies, we generated a transgenic line where EGFP expression is controlled by the zebrafish flk1 promoter. We find that angioblasts migrate as individual cells to form a vascular cord at the midline. This transient structure is stabilized by endothelial cell cell junctions, and subsequently undergoes lumen formation to form a fully patent vessel. Downregulating the VEGF signaling pathway, while affecting the number of angioblasts, does not appear to affect their migratory behavior. Our studies also indicate that the endoderm, a tissue previously implicated in vascular development, provides a substratum for endothelial cell migration and is involved in regulating the timing of this process, but that it is not essential for the direction of migration. In addition, the endothelial cells in endodermless embryos form properly lumenized vessels, contrary to what has been previously reported in Xenopus and avian embryos. These studies provide the tools and a cellular framework for the investigation of mutations affecting vasculogenesis in zebrafish. PMID- 16251213 TI - Cholinergic challenge in Alzheimer patients and mild cognitive impairment differentially affects hippocampal activation--a pharmacological fMRI study. AB - Pharmacological functional MRI (phMRI) examines the impact of pharmacologically induced neurochemical changes on brain function at a system level. The current phMRI study directly compared effects of cholinergic stimulation on brain function between patients with Alzheimer's disease and mild cognitive impairment, a disease stage preceding the development of Alzheimer's disease. Brain function during recognition of (un)familiar information was examined for changes after exposure to galantamine, a cholinesterase inhibitor used for treating memory deficits in Alzheimer's disease. Alzheimer patients [n = 18; age 74.5 years +/- 8.2; Mini-Mental State Examination (MMSE) 22.5 +/- 2.4] and patients with mild cognitive impairment (n = 28; mean age 73.6 +/- 7.5; MMSE 27.0 +/- 1.2) were scanned during face recognition under three different conditions: at baseline, and after acute (single dose) and prolonged exposure (5 days) to galantamine. Functional data were analysed in an event-related fashion. In both groups, acute exposure produced strong increases in brain activation (Z > 3.1). Prolonged exposure produced less strong effects that mainly involved decreases in activation (Z > 3.1). In mild cognitive impairment, acute exposure increased activation in posterior cingulate, left inferior parietal, and anterior temporal lobe. Prolonged exposure decreased activation in similar posterior cingulate areas, and in bilateral prefrontal areas. Effects were stronger for positive ('familiar') than for negative ('unfamiliar') decisions, indicating that the effect was specific to memory retrieval. In Alzheimer patients, acute exposure increased activation bilaterally in hippocampal areas, whereas prolonged exposure decreased activation in these areas. Effects were more pronounced for negative than for positive decisions, suggesting a preferential effect on memory encoding. Unique profiles of signal reactivity were found in a number of areas, including left inferior parietal lobe and left hippocampus proper. The reactivity of posterior cingulate and hippocampal structures to cholinergic challenge suggests a key role of the cholinergic system in the functional processes that lead to Alzheimer's disease. The differential response to cholinergic challenge in mild cognitive impairment and Alzheimer patients may reflect a difference in the functional status of the cholinergic system between both groups, which is in line with recent results showing a differential clinical response to cholinergic treatment. PMID- 16251215 TI - Type and frequency of mutations in the LRRK2 gene in familial and sporadic Parkinson's disease*. PMID- 16251214 TI - Reduced brain functional reserve and altered functional connectivity in patients with multiple sclerosis. AB - Cognitive dysfunction (affecting particularly attention and working memory) occurs early in patients with multiple sclerosis. Previous studies have focused on identifying potentially adaptive functional reorganization through recruitment of new brain regions that could limit expression of these deficits. However, lesion studies remind us that functional specializations in the brain make certain brain regions necessary for a given task. We therefore have asked whether altered functional interactions between regions normally recruited provide an alternative adaptive mechanism with multiple sclerosis pathology. We used a version of the n-back task to probe working memory in patients with early multiple sclerosis. We applied a functional connectivity analysis to test whether relationships between relative activations in different brain regions change in potentially adaptive ways with multiple sclerosis. We studied 21 patients with relapsing-remitting multiple sclerosis and 16 age- and sex-matched healthy controls with 3T functional MRI. The two groups performed equally well on the task. Task-related activations were found in similar regions for patients and controls. However, patients showed relatively reduced activation in the superior frontal and anterior cingulate gyri (P > 0.01). Patients also showed a variable, but generally substantially smaller increase in activation than healthy controls with greater task complexity, depending on the specific brain region assessed (P < 0.001). Functional connectivity analysis defined further differences not apparent from the univariate contrast of the task-associated activation patterns. Control subjects showed significantly greater correlations between right dorsolateral prefrontal and superior frontal/anterior cingulate activations (P < 0.05). Patients showed correlations between activations in the right and left prefrontal cortices, although this relationship was not significant in healthy controls (P < 0.05). We interpret these results as showing that, while cognitive processing in the task appears to be performed using similar brain regions in patients and controls, the patients have reduced functional reserve for cognition relevant to memory. Functional connectivity analysis suggests that altered inter hemispheric interactions between dorsal and lateral prefrontal regions may provide an adaptive mechanism that could limit clinical expression of the disease distinct from recruitment of novel processing regions. Together, these results suggest that therapeutic enhancement of the coherence of interactions between brain regions normally recruited (functional enhancement), as well as recruitment of alternative areas or use of complementary cognitive strategies (both forms of adaptive functional change), may limit expression of cognitive impairments in multiple sclerosis. PMID- 16251216 TI - SCA28, a novel form of autosomal dominant cerebellar ataxia on chromosome 18p11.22-q11.2. AB - We describe a four-generation Italian family with a novel form of juvenile-onset, slowly progressive, autosomal dominant cerebellar ataxia. Eleven affected family members have been evaluated. The mean age at onset was 19.5 years with no evidence of anticipation. The first symptoms were invariably unbalanced standing and mild gait incoordination. Gaze-evoked nystagmus was prominent at onset, while patients with longer disease duration developed slow saccades, ophthalmoparesis and, often, ptosis. Deep tendon reflexes in lower limbs were increased in 80% of the cases. Genetic analysis excluded the presence of pathological repeat expansions in spinocerebellar ataxia (SCA) types 1-3, 6-8, 10, 12 and 17, and DRPLA genes. Linkage exclusion tests showed no evidence of association with other known SCA loci. A genome-wide screen analysis identified linkage with chromosome 18 markers. A maximum two-point limit of determination score of 4.20 was found for marker D18S53. Haplotype analysis refined a critical region of 7.9 Mb between markers D18S1418 and D18S1104. This new SCA locus on 18p11.22-q11.2 has been designated SCA28. Candidate genes within the critical interval are currently screened for mutations. PMID- 16251218 TI - Olfactory ensheathing glial co-grafts improve functional recovery in rats with 6 OHDA lesions. AB - Olfactory ensheathing cells (OEC) transplanted to the site of a spinal cord injury can promote axonal sparing/regeneration and functional recovery. The purpose of this study was to investigate if OEC enhance the effects of grafted dopamine-neuron-rich ventral mesencephalic tissue (VM) in a rodent model of Parkinson's disease. We co-grafted VM with either OEC or astrocytes derived from the same olfactory bulbs as the OEC to rats with a unilateral 6-hydroxydopamine lesion of the nigrostriatal system. Co-grafting fetal VM with OEC, but not with astrocytes enhanced dopamine cell survival, striatal reinnervation and functional recovery of amphetamine- and apomorphine-induced rotational behaviour compared with grafting embryonic VM alone. Grafting OEC or astrocytes alone had no effects. Intriguingly, only in the presence of OEC co-grafts, did dopamine neurons extend strikingly long neurites that reached peripheral striatal compartments. Comparable results were observed in a co-culture system where OEC promoted dopamine cell survival and neurite elongation through a mechanism involving both releasable factors and direct contact. Cell type analysis of fetal VM grafts suggested that dopamine neurons of the substantia nigra rather than of the ventral tegmental area were increased in the presence of OEC co-grafts. We conclude that the addition of OEC enhances efficacy of grafted immature dopamine neurons in a rat Parkinson's disease model. PMID- 16251217 TI - Rhinal-hippocampal connectivity determines memory formation during sleep. AB - Compared with waking state attention, volition and semantic processing play a minor role during sleep. Thus, investigating declarative memory formation during sleep may allow us to isolate mnemonic core processes. The most feasible approach to memory formation during sleep is the analysis of dream memories. Lesion and imaging studies have demonstrated that encoding of declarative memories, i.e. consciously accessible events and facts, depends on operations within the rhinal cortex and the hippocampus, two substructures of the medial temporal lobe. Successful memory formation is accompanied by a transient rhinal-hippocampal interaction. Consequently, the ability to memorize dreams may be related to mediotemporal connectivity. Therefore, we recorded EEG during sleep from rhinal and hippocampal depth electrodes implanted in 12 epilepsy patients (eight women, mean age 41.1 +/- 6.4 years). They were awakened during rapid eye movement sleep (REM) and asked to recall their dream. Via coherence analyses we show that rhinal hippocampal connectivity values are approximately twice as large for patients with good dream recall versus those patients with poor recall. This suggests that rhinal-hippocampal connectivity is a key factor in determining declarative memory formation. PMID- 16251219 TI - IL-15 and IL-15R in leucocytes from patients with systemic lupus erythematosus. AB - OBJECTIVE: To assess the functional status of the IL-15/IL-15Ralpha cytokine system in different leucocyte subsets from patients with systemic lupus erythematosus (SLE). METHODS: Eighteen patients with SLE (10 with inactive and eight with active disease) and 14 healthy individuals were studied. Serum levels and in vitro production of IL-15 were determined. In addition, the expression of IL-15 receptor alpha (IL-15Ralpha) and membrane-bound IL-15 was assessed and the in vitro effects of IL-15 on CD69 and CD64 expression, interferon-gamma and TNF alpha synthesis, respiratory burst induction and apoptosis were studied. RESULTS: Serum levels of IL-15 were significantly increased in inactive and active patients with SLE. Accordingly, the in vitro synthesis and release of IL-15 by monocytes in response to IFN-gamma+lipopolysaccharide was significantly enhanced in SLE patients with active disease, as was the percentage of membrane-bound IL 15+ monocytes. On the other hand, enhanced basal expression of IL-15Ralpha was detected in leucocytes from SLE patients, with defective induction upon stimulation with phytohaemagglutinin or phorbol myristate acetate/ionomycin. Furthermore, diminished induction of CD69 expression and interferon-gamma and TNF alpha synthesis by recombinant human IL-15 was detected in peripheral blood mononuclear cells from SLE, and there was defective induction of CD64 and priming for respiratory burst in neutrophils. The anti-apoptotic effect of IL-15 was diminished in leucocytes from SLE patients. CONCLUSION: Our data indicate that there is enhanced synthesis of IL-15 by immune cells from SLE patients, with a poor response to this cytokine by different leucocyte subsets. This abnormal function of IL-15/IL-15Ralpha may contribute significantly to the pathogenesis of SLE. PMID- 16251220 TI - Regio- and enantioselectivities in epoxide conjugations are modulated by residue 210 in Mu class glutathione transferases. AB - The homologous human glutathione transferases (GSTs) M1-1 and M2-2 have similar catalytic activities with many electrophilic substrates, but differ strikingly in their conjugation of epoxides with glutathione. Residue 210, Thr in GST M2-2 and Ser in GST M1-1, is a key active-site component in determining the activity profile with epoxide substrates. This residue is hypervariable in Mu class GSTs, suggesting that it has special significance in the evolution of new functions. The present study shows that minor modifications of this residue can have major consequences for the enzyme-catalyzed epoxide conjugations. In general, a Ser at position 210 gives the highest catalytic efficiency, but the relatively high activity with an Ala placed on this position demonstrates that a hydroxyl group is not required. In contrast, a Thr residue suppresses the activity with epoxides by several orders of magnitude without major effects on the activity with alternative GST substrates. Residue 210 influences both the regio- and enantioselectivity with chiral and prochiral epoxides of stilbene and styrene and influences the distribution of isomeric glutathione conjugates. Thus, residue 210 contributes to both stereoselective recognition of the substrates and to partitioning of the isomeric reactants to the alternative transition states leading to separate chiral products. PMID- 16251221 TI - Generation of GPI-linked CCL5 based chemokine receptor antagonists for the suppression of acute vascular damage during allograft transplantation. AB - Limiting the acute vascular damage associated with leukocyte infiltration is a central issue in solid organ transplantation. The family of chemotactic cytokines (chemokines) helps to regulate leukocyte recruitment. Systemic treatment with the chemokine ligand-5 (CCL5) based antagonist Met-RANTES has previously shown to suppress acute damage to transplanted kidneys by blocking effector cell recruitment. To address problems associated with systemic long-term administration of chemokine receptor antagonists, a chemokine based reagent was designed to be integrated into endothelial surfaces of the organ just before transplantation. Proteins anchored by glycosylphosphatidylinositol (GPI), when purified and added to cells, are efficiently incorporated into their cell surface membranes. A series of modifications were introduced into the CCL5 protein to generate a functional antagonist. These included the addition of an N-terminal methionine group, a mutation to render the protein a dimer and a GPI signal sequence for surface expression. The resultant protein was stably expressed in CHO cells, GPI anchorage was confirmed and the protein purified by FPLC. Exogenously administered Met-CCL5(dimer)-GPI was efficiently inserted into the membrane of microvascular endothelial cells. The reagent is being tested in murine models of renal transplantation. The effect on subsequent immune induced damage will be assessed. PMID- 16251222 TI - The role of extra-membranous inter-helical loops in helix-helix interactions. AB - The effect of a short loop connecting two transmembrane alpha-helices was studied using molecular dynamics simulations. Helices F and G from bacteriorhodopsin and two corresponding polyalanine helices were embedded in octane and POPC membranes in a transmembrane configuration both with and without the inter-helical loop. The results indicate that the membrane environment and the sequence of the loop are more influential on the dynamics and structure of the motif than the presence of a loop as such, at least for the time-scales investigated. The four residues in the FG loop are stabilized by four hydrogen bonds. These hydrogen bonds are not present in the polyalanine loop, causing it to be more flexible than the FG loop. This effect was observed independently of the protein environment, stressing the importance of the sequence. The structural analysis indicates that the loop has weak stabilizing properties in all environments. The stabilization due to the presence of the loop was strongest in a simulation of the FG fragment in a membrane-mimetic octane slab. In the simulations of the helix-loop-helix motif embedded in an explicit lipid bilayer model, the lipid bilayer interface compensates to a large extent for the absence of the loop. PMID- 16251223 TI - Young people and healthy eating: a systematic review of research on barriers and facilitators. AB - A systematic review was conducted to examine the barriers to, and facilitators of, healthy eating among young people (11-16 years). The review focused on the wider determinants of health, examining community- and society-level interventions. Seven outcome evaluations and eight studies of young people's views were included. The effectiveness of the interventions was mixed, with improvements in knowledge and increases in healthy eating but differences according to gender. Barriers to healthy eating included poor school meal provision and ease of access to, relative cheapness of and personal taste preferences for fast food. Facilitators included support from family, wider availability of healthy foods, desire to look after one's appearance and will power. Friends and teachers were generally not a common source of information. Some of the barriers and facilitators identified by young people had been addressed by soundly evaluated effective interventions, but significant gaps were identified where no evaluated interventions appear to have been published (e.g. better labelling of food products), or where there were no methodologically sound evaluations. Rigorous evaluation is required particularly to assess the effectiveness of increasing the availability of affordable healthy food in the public and private spaces occupied by young people. PMID- 16251224 TI - Non-invasive half millimetre 32 detector row computed tomography angiography accurately excludes significant stenoses in patients with advanced coronary artery disease and high calcium scores. AB - OBJECTIVE: To show an overall diagnostic accuracy > or = 90% for detection of > or = 50% stenoses by coronary half millimetre 32 detector row computed tomography angiography (32 x 0.5-MDCTA) in patients with advanced coronary artery disease (CAD) and a high likelihood of raised calcium scores. METHODS: ECG gated 32 x 0.5 MDCTA (32 x 0.5 mm cross sections, 0.35 x 0.35 x 0.35 mm3 isotropic voxels, 400 ms rotation) was performed after injection of iodixanol (120 ml, 320 mg/ml) in 30 consecutive patients (25 men, mean (SD) age 59 (13) years, body mass index 26.2 (4.9) kg/m2). Native arteries, including > or = 1.5 mm branches, and bypass grafts were screened for > or = 50% stenoses. Stents were excluded. Conventional coronary angiography (performed 18 (12) days before 32 x 0.5-MDCTA) was analysed by quantitative coronary angiography. RESULTS: Median Agatston calcium score was 510 (range 3-5066). Sensitivity, specificity, and positive and negative predictive values for detection of > or = 50% stenoses in native arteries were 76% (29 of 38), 94% (190 of 202), 71% (29 of 41), and 96% (190 of 199), respectively. Overall diagnostic accuracy was 91% (219 of 240). Due to the following artefacts 20% (69 of 352) of the vessels were excluded: motion, noise, and low contrast enhancement isolated or in combination (45 of 69 (65%)); image distortion by implantable cardioverter-defibrillator or pacemaker leads (18 of 69 (26%)); and blooming secondary to severe calcification (6 of 69 (9%)). CONCLUSIONS: Coronary 32 x 0.5-MDCTA accurately excludes > or = 50% stenoses in patients with advanced CAD and high calcium scores with an overall diagnostic accuracy of 91%. PMID- 16251227 TI - Natriuretic peptides in heart valve disease. AB - Synthesis and release of B-type natriuretic peptide (BNP) are increased in heart failure, and plasma concentrations provide important therapeutic and prognostic information. Recent studies have shown that BNP concentrations are also increased with disease of the mitral and aortic valves. The extent of the increase is broadly related to the severity of the valve abnormality and the degree of consequent cardiac remodelling. BNP concentrations appear to relate to prognosis in these patients and might have a role in identifying suitable candidates for cardiac surgery. This paper reviews the current literature and identifies areas where further research is required if assessment of BNP is to be of practical use. PMID- 16251226 TI - Impact of intracoronary aspiration thrombectomy during primary angioplasty on left ventricular remodelling in patients with anterior ST elevation myocardial infarction. AB - OBJECTIVE: To evaluate prospectively the impact on left ventricular (LV) remodelling of an intracoronary aspiration thrombectomy device as adjunctive therapy in primary percutaneous coronary intervention (PCI) in patients with anterior ST elevation myocardial infarction (STEMI). METHODS: 76 consecutive patients with anterior STEMI (65.3 (11.2) years, 48 men) were randomly assigned to intracoronary thrombectomy and stent placement (n = 38) or to conventional stenting (n = 38) of the infarct related artery. Each patient underwent transthoracic echocardiography immediately after PCI and at six months. At the time of echocardiographic control, major adverse cardiovascular events (MACE) in terms of death, new onset of myocardial infarction, and hospitalisation for heart failure were also evaluated. RESULTS: After a successful primary PCI, patients in the thrombectomy group achieved a higher rate of post-procedure myocardial blush grade 3 (36.8% v 13.1%, p = 0.03) and effective ST segment resolution at 90 minutes (81.6% v 55.3%, p = 0.02). Six months after the index intervention, 19 patients (26.8%) developed LV dilatation, defined as an increase in end diastolic volume (EDV) >or= 20%: 15 in the conventional group and four in the thrombectomy group (p = 0.006). Accordingly, at six months patients treated conventionally had significantly higher end systolic volumes (82 (7.7) ml v 75.3 (4.9) ml, p < 0.0001) and EDV (152.5 (18.1) ml v 138.1 (10.7) ml, p < 0.0001) than patients treated with thrombectomy. No differences in cumulative MACE were observed (10.5% in the conventional group v 8.6% in the thrombectomy group, not significant). CONCLUSION: Compared with conventional stenting, adjunctive aspiration thrombectomy in successful primary PCI seems to be associated with a significantly lower incidence of LV remodelling at six months in patients with anterior STEMI. PMID- 16251225 TI - Mitral repair best practice: proposed standards. AB - OBJECTIVES: To define best practice standards for mitral valve repair surgery. DESIGN: Development of standards for process and outcome by consensus. SETTING: Multidisciplinary panel of surgeons, anaesthetists, and cardiologists with interests and expertise in caring for patients with severe mitral regurgitation. MAIN OUTCOME MEASURES: Standards for best practice were defined including the full spectrum of multidisciplinary aspects of care. RESULTS: 19 criteria for best practice were defined including recommendations on surgical training, intraoperative transoesophageal echocardiography, surgery for atrial fibrillation, audit, and cardiology and imaging issues. CONCLUSIONS: Standards for best practice in mitral valve repair were defined by multidisciplinary consensus. This study gives centres undertaking mitral valve repair an opportunity to benchmark their care against agreed standards that are challenging but achievable. Working towards these standards should act as a stimulus towards improvements in care. PMID- 16251228 TI - Admission N-terminal pro-brain natriuretic peptide and its interaction with admission troponin T and ST segment resolution for early risk stratification in ST elevation myocardial infarction. AB - OBJECTIVE: To assess the long term prognostic value of N-terminal pro-brain natriuretic peptide (NT-proBNP) on admission and its prognostic interaction with both admission troponin T (TnT) concentrations and resolution of ST segment elevation in fibrinolytic treated ST elevation myocardial infarction (STEMI). DESIGN AND SETTING: Substudy of the ASSENT (assessment of the safety and efficacy of a new thrombolytic) -2 and ASSENT-PLUS trials. PATIENTS: NT-proBNP and TnT concentrations were determined on admission in 782 patients. According to NT proBNP concentrations, patients were divided into three groups: normal concentration (for patients < or = 65 years, < or = 184 ng/l and < or = 268 ng/l and for those > 65 years, < or = 269 ng/l and < or = 391 ng/l in men and women, respectively); higher than normal but less than the median concentration (742 ng/l); and above the median concentration. For TnT, a cut off of 0.1 microg/l was used. Of the 782 patients, 456 had ST segment resolution (< 50% or > or = 50%) at 60 minutes calculated from ST monitoring. MAIN OUTCOME MEASURES: All cause one year mortality. RESULTS: One year mortality increased stepwise according to increasing concentrations of NT-proBNP (3.4%, 6.5%, and 23.5%, respectively, p < 0.001). In receiver operating characteristic analysis, NT-proBNP strongly trended to be associated more with mortality than TnT and time to 50% ST resolution (area under the curve 0.81, 95% confidence interval (CI) 0.72 to 0.9, 0.67, 95% CI 0.56 to 0.79, and 0.66, 95% CI 0.56 to 0.77, respectively). In a multivariable analysis adjusted for baseline risk factors and TnT, both raised NT-proBNP and ST resolution < 50% were independently associated with higher one year mortality, whereas raised TnT contributed independently only before information on ST resolution was added to the model. CONCLUSION: Admission NT-proBNP is a strong independent predictor of mortality and gives, together with 50% ST resolution at 60 minutes, important prognostic information even after adjustment for TnT and baseline characteristics in STEMI. PMID- 16251230 TI - Changes in circulating mesenchymal stem cells, stem cell homing factor, and vascular growth factors in patients with acute ST elevation myocardial infarction treated with primary percutaneous coronary intervention. AB - OBJECTIVE: To investigate the spontaneous occurrence of circulating mesenchymal stem cells (MSC) and angiogenic factors in patients with ST elevation acute myocardial infarction (STEMI) treated with primary percutaneous coronary intervention (PCI). DESIGN: In 20 patients with STEMI, blood samples were obtained on days 1, 3, 7, 14, 21, and 28 after the acute PCI. Fifteen patients with a normal coronary angiography formed a control group. MSC (CD45-/CD34-), plasma stromal derived factor 1 (SDF-1), vascular endothelial growth factor A (VEGF-A), and fibroblast growth factor 2 (FGF-2) were measured by multiparametric flow cytometry and enzyme linked immunosorbent assay (ELISA). RESULTS: Circulating CD45-/CD34- cells were significantly decreased on day 7 compared with day 3. Cell counts normalised one month after the acute onset of STEMI. The changes were mainly seen in patients with a large infarction. Plasma SDF-1 increased significantly from day 3 to day 28, and VEGF-A and FGF-2 increased significantly from day 7 to day 28. CONCLUSIONS: Spontaneous sequential fluctuations in MSC and the increase in vascular growth factor concentrations after STEMI suggest that the optimal time for additional stem cell therapy is three weeks after a myocardial infarction to obtain the maximum effects by stimulating endogenous growth factors on the delivered stem cells. PMID- 16251229 TI - Efficacy of drug eluting stents in patients with and without diabetes mellitus: indirect comparison of controlled trials. AB - OBJECTIVE: To examine whether polymer based coronary stents eluting sirolimus or paclitaxel are equally effective in patients with and without diabetes. METHODS: Systematic review and meta-analysis by indirect comparison of randomised controlled trials comparing stents eluting sirolimus or paclitaxel with conventional bare metal stents. The overall study population and patients with and without diabetes were analysed separately by using the ratio of incidence rate ratios (RIRR). RESULTS: The analysis was based on 10 trials (six with sirolimus, four with paclitaxel), 4513 patients (1146 patients with diabetes), 5755 years of follow up, and 2464 events. In patients without diabetes sirolimus eluting stents were superior to paclitaxel eluting stents with respect to in stent (RIRR 0.21, 95% confidence interval (CI) 0.10 to 0.48, p < 0.001) and in segment restenosis (RIRR 0.47, 95% CI 0.24 to 0.92, p = 0.027), target lesion revascularisation (RIRR 0.54, 95% CI 0.30 to 0.99, p = 0.045), and major adverse cardiac events (RIRR 0.46, 95% CI 0.26 to 0.83, p = 0.010). In patients with diabetes the two drug eluting stents did not differ significantly in any of these end points. Meta-regression analysis showed a significant difference between patients with and without diabetes (tests for interaction for in-stent and in segment restenosis, p = 0.036 and p = 0.016). CONCLUSION: Indirect evidence indicates that sirolimus eluting stents are superior to paclitaxel eluting stents in patients without diabetes but not in patients with diabetes. PMID- 16251231 TI - Prevention of cardiovascular diseases: focus on modifiable cardiovascular risk. AB - OBJECTIVE: To determine whether the use of a 20% absolute risk threshold for cardiovascular disease as recommended in current guidelines leads to exclusion of patients with a substantial modifiable risk (> or = 5%). METHODS: Data collected within the framework of a randomised controlled trial in three primary health care centres located in deprived neighbourhoods were analysed. The 10 year absolute risk and the modifiable part of risk were calculated by using the Framingham risk equation. Among patients with a modifiable risk reduction of > or = 5% (number needed to treat < or = 20) the characteristics and risk factors of patients with an absolute risk > or = 20% and those with an absolute risk < 20% were compared. RESULTS: 293 patients aged 30-70 years at risk of developing cardiovascular disease were included, of whom 66% were women and 36% were of Dutch origin. Of all patients, 33% had an absolute risk > or = 20% and 61% had a modifiable risk > or = 5%. Of those at > or = 20% absolute risk, a vast majority (98%) had a modifiable risk > or = 5%. Among those with an absolute risk < 20%, 43% had a modifiable risk > or = 5%; this group, who were relatively young and predominantly women, constituted 29% of the entire study population. CONCLUSIONS: Targeting preventive strategies at a 10 year absolute risk > or = 20% leads to exclusion of a large group of relatively young, predominantly female patients. In total, about one quarter had an absolute risk < 20% but a modifiable risk > or = 5% and should therefore benefit from intervention. PMID- 16251233 TI - Prognostic value of systolic mitral annular velocity measured with Doppler tissue imaging in patients with chronic heart failure caused by left ventricular systolic dysfunction. AB - OBJECTIVE: To assess the prognostic value of various conventional and novel echocardiographic indices in patients with chronic heart failure (CHF) caused by left ventricular (LV) systolic dysfunction. METHODS: 185 patients with a mean (SD) age of 67 (11) years with CHF and LV ejection fraction < 45% despite optimal pharmacological treatment were prospectively enrolled. The patients underwent two dimensional echocardiography with tissue harmonic imaging to assess global LV systolic function and obtain volumetric data. Transmitral flow was assessed with conventional pulse wave Doppler. Systolic (Sm), early, and late diastolic mitral annular velocities were measured with the use of colour coded Doppler tissue imaging. RESULTS: During a median follow up of 32 months (range 24-38 months in survivors), 34 patients died and one underwent heart transplantation. Sm velocity (hazard ratio (HR) 0.648, 95% confidence interval (CI) 0.463 to 0.907, p = 0.011), diastolic arterial pressure (HR 0.965, 95% CI 0.938 to 0.993, p = 0.015), serum creatinine (HR 1.006, 95% CI 1.001 to 1.011, p = 0.023), LV ejection fraction (HR 0.945, 95% CI 0.899 to 0.992, p = 0.024), age (HR 1.035, 95% CI 1.000 to 1.071, p = 0.052), LV end systolic volume index (HR 1.009, 95% CI 0.999 to 1.019, p = 0.067), and restrictive pattern of transmitral flow (HR 0.543, 95% CI 0.278 to 1.061, p = 0.074) predicted the outcome of death or transplantation on univariate analysis. On multivariate analysis, only Sm velocity (HR 0.648, 95% CI 0.460 to 0.912, p = 0.013) and diastolic arterial pressure (HR 0.966, 95% CI 0.938 to 0.994, p = 0.016) emerged as independent predictors of outcome. CONCLUSIONS: In patients with CHF and LV systolic dysfunction despite optimal pharmacological treatment, the strongest independent echocardiographic predictor of prognosis was Sm velocity measured with quantitative colour coded Doppler tissue imaging. PMID- 16251234 TI - Autotopes and allotopes. PMID- 16251232 TI - Prosthesis-patient mismatch: definition, clinical impact, and prevention. AB - Prosthesis-patient mismatch (PPM) is present when the effective orifice area of the inserted prosthetic valve is too small in relation to body size. Its main haemodynamic consequence is to generate higher than expected gradients through normally functioning prosthetic valves. This review updates the present knowledge about the impact of PPM on clinical outcomes. PPM is common (20-70% of aortic valve replacements) and has been shown to be associated with worse haemodynamic function, less regression of left ventricular hypertrophy, more cardiac events, and lower survival. Moreover, as opposed to most other risk factors, PPM can largely be prevented by using a prospective strategy at the time of operation. PMID- 16251235 TI - Effect of pravastatin in people with diabetes and chronic kidney disease. AB - Although diabetes is a major cause of chronic kidney disease (CKD), limited data describe the cardiovascular benefit of hydroxymethyl glutaryl CoA reductase inhibitors (statins) in people with both of these conditions. This study sought to determine whether pravastatin reduced the incidence of first or recurrent cardiovascular events in people with non-dialysis-dependent CKD and concomitant diabetes, using data from three randomized trials of pravastatin 40 mg daily versus placebo. CKD was defined by estimated GFR <60 or 60 to 89.9 ml/min per 1.73 m2 with proteinuria. Of 19,737 patients, 4099 (20.8%) had CKD but not diabetes at baseline, 873 (4.4%) had diabetes but not CKD, and 571 (2.9%) had both conditions. The primary composite outcome was time to myocardial infarction, coronary death, or percutaneous/surgical coronary revascularization. Median follow-up was 64 mo. After adjustment for trial and random treatment assignment, the incidence of the primary outcome was lowest in individuals with neither CKD nor diabetes (15.2%), intermediate in individuals with only CKD (18.6%) or only diabetes (21.3%), and highest in individuals with both characteristics (27.0%). Pravastatin reduced the relative likelihood of the primary outcome to a similar extent in subgroups defined by the presence or absence of CKD and diabetes. For example, pravastatin was associated with a significant reduction in the relative risk of the primary outcome by 25% in patients with CKD and concomitant diabetes and by 24% in individuals with neither characteristic. However, the absolute reduction in the risk of the primary outcome as a result of pravastatin use was highest in patients with both CKD and diabetes (6.4%) and lowest in individuals with neither characteristic (3.5%). In conclusion, stage 2 or early stage 3 CKD and diabetes both are associated with higher cardiovascular risk, and pravastatin reduces cardiovascular event rates in people with neither, one, or both characteristics. Given the high absolute benefit of pravastatin in patient with diabetes and stage 2 or early stage 3 CKD, this population in particular should be targeted for widespread use of statins. Additional studies are needed to determine whether these benefits apply to patients with more severe CKD, and recruitment to such studies should be given high priority. PMID- 16251236 TI - Mutational and Biological Analysis of alpha-actinin-4 in focal segmental glomerulosclerosis. AB - Mutations in the alpha-actinin-4 gene (ACTN4) cause an autosomal dominant form of focal segmental glomerulosclerosis (FSGS). A mutational analysis was performed of ACTN4 in DNA from probands with a family history of FSGS as well as in individuals with nonfamilial FSGS. The possible contribution of noncoding variation in ACTN4 to the development of FSGS also was assessed. Multiple nucleotide variants were identified in coding and noncoding sequence. The segregation of nonsynonymous coding sequence variants was examined in the relevant families. Only a small number of nucleotide changes that seemed likely to be causing (or contributing to) disease were identified. Sequence changes that predicted I149del, W59R, V801M, R348Q, R837Q, and R310Q changes were identified. For studying their biologic relevance and their potential roles in the pathogenesis of FSGS, these variants were expressed as GFP-fusion proteins in cultured podocytes. F-actin binding assays also were performed. Three of these variants (W59R, I149del, and V801M) showed clear cellular mislocalization in the form of aggregates adjacent to the nucleus. Two of these mislocalized variants (W59R and I149del) also showed an increased actin-binding activity. The I149del mutation segregated with disease; W59R was found to be a de novo mutation in the proband. A total of five ACTN4 mutations that are believed to be disease causing (three reported previously and two novel) as well as a number of variants with unclear contribution to disease now have been identified. The possibility that some of these other variants increase the susceptibility to FSGS cannot be excluded. ACTN4 mutations seem to account for approximately 4% of familial FSGS. PMID- 16251237 TI - Uric acid-induced C-reactive protein expression: implication on cell proliferation and nitric oxide production of human vascular cells. AB - Recent experimental and human studies have shown that hyperuricemia is associated with hypertension, systemic inflammation, and cardiovascular disease mediated by endothelial dysfunction and pathologic vascular remodeling. Elevated levels of C reactive protein (CRP) have emerged as one of the most powerful independent predictors of cardiovascular disease. In addition to being a marker of inflammation, recent evidence suggests that CRP may participate directly in the development of atherosclerotic vascular disease. For investigating whether uric acid (UA)-induced inflammatory reaction and vascular remodeling is related to CRP, the UA-induced expression of CRP in human vascular smooth muscle cells (HVSMC) and human umbilical vein endothelial cells (HUVEC) was examined, as well as the pathogenetic role of CRP in vascular remodeling. It is interesting that HVSMC and HUVEC expressed CRP mRNA and protein constitutively, revealing that vascular cells are another source of CRP production. UA (6 to 12 mg/dl) upregulated CRP mRNA expression in HVSMC and HUVEC with a concomitant increase in CRP release into cell culture media. Inhibition of p38 or extracellular signal regulated kinase 44/42 significantly suppressed UA-induced CRP expression, implicating these pathways in the response to UA. UA stimulated HVSMC proliferation whereas UA inhibited serum-induced proliferation of HUVEC assessed by 3H-thymidine uptake and cell counting, which was attenuated by co-incubation with probenecid, the organic anion transport inhibitor, suggesting that entry of UA into cells is responsible for CRP expression. UA also increased HVSMC migration and inhibited HUVEC migration. In HUVEC, UA reduced nitric oxide (NO) release. Treatment of vascular cells with anti-CRP antibody revealed a reversal of the effect of UA on cell proliferation and migration in HVSMC and NO release in HUVEC, which suggests that CRP expression may be responsible for UA-induced vascular remodeling. This is the first study to show that soluble UA, at physiologic concentrations, has profound effects on human vascular cells. The observation that UA alters the proliferation/migration and NO release of human vascular cells, mediated by the expression of CRP, calls for careful reconsideration of the role of UA in hypertension and vascular disease. PMID- 16251238 TI - Expression of surfactant protein-C, S100A8, S100A9, and B cell markers in renal allografts: investigation of the prognostic value. AB - The intent of this study was to identify genes of which expression during acute rejection is associated with progression to chronic allograft nephropathy using gene expression profiling. Ten patients who had graft loss through chronic allograft nephropathy (progression [PR] group) and 18 patients who had stable graft function over time (nonprogression [NP] group) were studied. Rejection severity and extent of infiltrating leukocytes in acute rejection biopsies were similar for both groups. Microarray analysis and real-time PCR validation showed that surfactant protein-C (SP-C), S100 calcium-binding protein A8 (S100A8), S100A9, and beta-globin levels distinguished the two groups. Relationship between expression of B cell markers and prognosis was also examined. Location in the graft of the protein and mRNA expression of candidate genes was investigated. The prognostic value of mRNA transcripts was tested in an independent cohort of 43 rejection biopsies. mRNA and protein expression of S100A8 and S100A9 in infiltrating cells was significantly higher in the NP group compared with the PR group. Expression of SP-C was four-fold higher in the PR group and was detected in glomeruli. No association between B cell clusters and outcome was found. In the second group of acute rejection biopsies, SP-C mRNA levels predicted renal function course beyond 6 mo in multivariate analysis. Relatively high expression of S100A8 and S100A9 during acute rejection is associated with a favorable prognosis, and high SP-C expression is associated with an unfavorable prognosis. Messenger RNA transcripts complement the biopsy in the prediction of graft function deterioration. PMID- 16251239 TI - Kidney function as a predictor of noncardiovascular mortality. AB - Chronic kidney disease is associated with a higher risk for cardiovascular mortality, as well as all-cause mortality. Whether chronic kidney disease is a predictor of noncardiovascular mortality is less clear. To further explore the latter, the association of kidney function with total noncardiovascular mortality and cause-specific mortality was assessed in the Cardiovascular Health Study, a community-based cohort of older individuals. Kidney disease was assessed using cystatin C and estimated GFR in 4637 participants in 1992 to 1993. Participants were followed until June 30, 2001. Deaths were adjudicated as cardiovascular or noncardiovascular disease by committee, and an underlying cause of death was assigned. The associations of kidney function with total noncardiovascular mortality and cause-specific mortality were analyzed by proportional hazards regression. Noncardiovascular mortality rates increased with higher cystatin C quartiles (16.8, 17.1, 21.6, and 50.0 per 1000 person-years). The association of cystatin C with noncardiovascular mortality persisted after adjustment for demographic factors; the presence of diabetes, C-reactive protein, hemoglobin, and prevalent cardiovascular disease; and measures of atherosclerosis (hazard ratio 1.69; 95% confidence interval 1.33 to 2.15, for the fourth quartile versus the first quartile). Results for estimated GFR were similar. The risk for noncardiac deaths attributed to pulmonary disease, infection, cancer, and other causes was similarly associated with cystatin C levels. Kidney function predicts noncardiovascular mortality from multiple causes in the elderly. Further research is needed to understand the mechanisms and evaluate interventions to reduce the high mortality rate in chronic kidney disease. PMID- 16251240 TI - IL-10 suppresses chemokines, inflammation, and fibrosis in a model of chronic renal disease. AB - IL-10 is a pluripotent cytokine that plays a pivotal role in the regulation of immune and inflammatory responses. Whereas short-term administration of IL-10 has shown benefit in acute glomerulonephritis, no studies have addressed the potential benefits of IL-10 in chronic renal disease. Chronically elevated blood levels of IL-10 in rats were achieved by administration of a recombinant adeno associated virus serotype 1 IL-10 (rAAV1-IL-10) vector. Control rats were given a similar dose of rAAV1-GFP. Four weeks after injection, IL-10 levels in serum were measured by ELISA, and chronic renal disease was induced by a 5/6 nephrectomy (n = 6 in each group). Eight weeks later, rats were killed and renal tissue was obtained for RNA, protein, and immunohistochemical analysis. Serum levels of IL 10 were 12-fold greater in the rAAV1-IL-10 group by 4 wk after rAAV1-IL-10 administration (345 +/- 169 versus 28 +/- 15 pg/ml; P = 0.001), and levels were maintained throughout the experiment. rAAV1-IL-10 treatment resulted in less proteinuria (P < 0.05), lower serum creatinine (P < 0.05), and higher creatinine clearances (P < 0.01) compared with rAAV1-GFP-treated rats. Renal interstitial infiltration was significantly attenuated by rAAV1-IL-10 administration as assessed by numbers of CD4+, CD8+, monocyte-macrophages (ED-1+) and dendritic (OX 62+) cells (P < 0.05), and this correlated with reductions in the renal expression of monocyte (renal monocyte chemoattractant protein-1 mRNA and protein) and T cell (RANTES mRNA) chemokines. rAAV1-IL-10 administration decreased mRNA levels of IFN-gamma and IL-2 in the kidney. The reduction in inflammatory cells was associated with a significant reduction in glomerulosclerosis and interstitial fibrosis. It is concluded that IL-10 blocks inflammation and improves renal function in this model of chronic renal disease. The feasibility of long-term overexpression of a gene using the AAV serotype 1 vector system in a model of renal disease is also demonstrated. PMID- 16251241 TI - Kappa-opioid system in uremic pruritus: multicenter, randomized, double-blind, placebo-controlled clinical studies. AB - Uremic pruritus is a very common and frustrating condition for both patients and clinicians because no treatment has been demonstrated to be effective in relieving the itch. In this report, nalfurafine, a new kappa-opioid receptor agonist, was used to treat uremic pruritus in patients who were undergoing routine hemodialysis. Two multicenter, randomized, double-blind, placebo controlled studies enrolled 144 patients with uremic pruritus to postdialysis intravenous treatment with either nalfurafine or placebo for 2 to 4 wk. A meta analysis approach was used to assess the efficacy of nalfurafine. Statistically significant reductions in worst itching (P = 0.0212), itching intensity (P = 0.0410), and sleep disturbances (P = 0.0003) were noted in the nalfurafine group as compared with placebo. Improvements in itching (P = 0.0025) and excoriations (P = 0.0060) were noted for the nalfurafine-treated patients. Nalfurafine showed similar types and incidences of drug-related adverse events as did placebo. Nalfurafine was shown to be an effective and safe compound for use in this severely ill patient population. PMID- 16251242 TI - Extracellular ATP determines 11beta-hydroxysteroid dehydrogenase type 2 activity via purinergic receptors. AB - Hypertension and sodium retention are features of a diminished 11beta hydroxysteroid dehydrogenase type 2 (11beta-HSD2). The activity of this enzyme is reduced in various disease states with abnormal renal sodium retention and hypertension, including preeclampsia. ATP release to the extracellular compartment is observed with shear stress, inflammation, and placental ischemia. It was hypothesized that ATP downregulates 11beta-HSD2 activity. For that purpose, cell lines from different tissues that previously were used to study the regulation of 11beta-HSD2 were investigated: JEG-3, a vascular trophoblastic; LLCPK1, a renal tubular; and SW620, a colonic epithelial cell line. The 11beta HSD2 activity, assessed by the conversion of 3H-cortisol to cortisone, was reversibly reduced during incubation with ATP or its stable analogue ATPgammaS in intact JEG-3 and LLCPK1, but not in SW620 cells. In JEG-3 cells, the purinergic antagonist pyridoxalphosphate-6-azophenyl-2',4'-disulphonic acid but not suramin reversed the inhibition. Incubation with UTP and ADP and their degradation products including adenosine and alpha,beta-methylene-ATP did not inhibit 11beta HSD2 activity. In contrast, 11beta-HSD2 activity increased almost 2.5-fold after incubation with 2'-methylthio-ATP. This indicates a bidirectional regulation by nucleotides via purinergic receptors. In JEG-3 cells, ATP/ATPgammaS did not alter 11beta-HSD2 promoter activity but reduced 11beta-HSD2 protein and mRNA concentration and half-life, suggesting a posttranscriptional regulation. In conclusion, ATP inhibits cell type specifically via purinergic receptors the expression and activity of the 11beta-HSD2 by a posttranscriptional mechanism. PMID- 16251244 TI - Coronary heart disease in patients with chronic kidney disease. PMID- 16251243 TI - Tissue kallikrein-deficient mice display a defect in renal tubular calcium absorption. AB - Renal tubular calcium (RTCa) transport is one of the main factors that determine serum Ca concentration and urinary Ca excretion. The distal convoluted and connecting tubules reabsorb a significant fraction (10%) of filtered Ca. These tubule segments also synthesize in large abundance tissue kallikrein (TK), a major kinin-forming enzyme. Tested was the hypothesis that TK and kinins are involved in controlling RTCa transport by studying TK (TK-/-) or kinin B2 receptor (B2-/-)-deficient mice on different Ca diets. On a 0.9% wt/wt Ca diet, 129Sv or C57Bl/6 TK-/- mice excreted significantly more Ca in urine than their wild-type (WT) littermates. There was no difference between TK-/- and WT mice for plasma concentrations of Ca, Mg, creatinine, parathyroid hormone, or 1,25 dihydroxyvitamin D. On a low Ca (LCa) diet (0.01% wt/wt), urinary Ca excretion decreased in both TK-/- and WT mice but still remained higher in TK-/- mice compared with WT. The plasma Ca concentration was unchanged in C57Bl/6 TK-/- mice but decreased significantly in 129Sv TK-/- mice. Taken together, these data demonstrate that TK deficiency led to impaired RTCa absorption. On the LCa diet, renal TK gene expression doubled in WT mice. No change in urinary Ca excretion was observed in B2-/- mice, even after treatment with a kinin B1-receptor antagonist, and these mice adapted normally to the LCa diet. TK deficiency had no effect on the renal abundance of distal Ca transporter mRNA. These data suggest that TK may be a physiologic regulator of RTCa transport, acting through a non kinin-mediated mechanism. PMID- 16251245 TI - Control of hyperphosphatemia among patients with ESRD. AB - Derangements of mineral metabolism occur during the early stages of chronic kidney disease (CKD). Hyperphosphatemia develops in the majority of patients with ESRD and has long been associated with progression of secondary hyperparathyroidism and renal osteodystrophy. More recent observational data have associated hyperphosphatemia with increased cardiovascular mortality among dialysis patients. Adequate control of serum phosphorus remains a cornerstone in the clinical management of patients with CKD not only to attenuate the progression of secondary hyperparathyroidism but also possibly to reduce the risk for vascular calcification and cardiovascular mortality. These measures include dietary phosphorus restriction, dialysis, and oral phosphate binders. Dietary restriction is limited in advanced stages of CKD. Phosphate binders are necessary to limit dietary absorption of phosphorus. Aluminum hydroxide is an efficient binder; however, its use has been nearly eliminated because of concerns of toxicity. Calcium salts are inexpensive and have been used effectively worldwide as an alternative to aluminum. Concerns of calcium overload have led to the investigation of alternatives. Currently, only two Food and Drug Administration approved noncalcium, nonaluminum binders are available. Sevelamer hydrochloride is an exchange resin and was not as effective as calcium acetate in meeting new guideline recommendations in one double-blind clinical trial. Lanthanum carbonate is a rare earth element and has been studied less extensively. Concerns of long term administration and toxicity exist. Furthermore, these agents are significantly more expensive than calcium salts, which may contribute to patient noncompliance. PMID- 16251246 TI - Effect of lipid modification on progression of coronary calcification. AB - Coronary artery calcification (CAC) reflects the anatomic presence of coronary atherosclerosis and the relative burden of coronary artery disease (CAD). Higher levels of CAC are seen in the presence of CAD risk factors, older age, and chronic kidney disease. The lipid profile (primarily low HDL cholesterol, elevated triglycerides, elevated LDL cholesterol, and elevated total cholesterol) are important factors in the calcification process. The annual progression of CAC can be reduced from 25 to 30% to 0 to 6% with LDL cholesterol reduction caused by statins and possibly sevelamer. At treated LDL cholesterol levels somewhere below 100 mg/dl, several sources of data suggest the anatomic burden of CAD, including CAC, regresses. Additional supportive studies indicate that carotid intimal medial thickness and the volume of coronary atheroma also can be reduced by LDL cholesterol reduction in concert with elevation of HDL cholesterol. This article reviews the data in support of altering the natural history of CAC with lipid modification. PMID- 16251247 TI - Strategies for improving long-term survival in patients with ESRD. AB - In 2003, more than 320,000 people in the United States were receiving dialysis for ESRD, with predicted increases to 650,000 by 2010 and 2 million by 2030. Mortality from cardiovascular disease (CVD) in patients with ESRD is 10 to 30 times higher than in the general population. The exact mechanism of accelerated CVD in patients with kidney disease is unknown. Treatment costs for ESRD are in excess of $14 billion annually (6.4% of Medicare budget). Strategies to improve long-term outcomes include aggressive risk factor modification, minimization of dialysis complications, and kidney transplantation. Because abnormalities of mineral metabolism contribute to mortality risk, phosphate binder therapy is fundamental. More expensive non-calcium-containing phosphate binders such as sevelamer have been recommended to reduce cardiovascular calcification. However, the lack of outcome data and the $2 to $3 billion annual cost make it difficult to justify widespread utilization of newer binders as first-line therapy. Conversely, kidney transplantation is known to improve survival in ESRD. Progression of atherosclerosis and CVD in patients with renal failure is largely due to loss of renal function per se, and provision of a functioning kidney through renal transplantation halts the progression of CVD and dramatically reduces mortality. Despite this fact, many patients lose Medicare funding for immunosuppressive therapy 3 yr posttransplantation. To achieve the goal of prevention of cardiovascular mortality in patients with ESRD, it clearly would be more prudent, efficacious, and cost-effective to use Medicare prescription drug dollars to provide full coverage for life-long immunosuppressive drug therapy after renal transplantation. PMID- 16251248 TI - Protecting calcium and phosphate balance in chronic renal disease. PMID- 16251249 TI - Reducing the burden of cardiovascular calcification in patients with chronic kidney disease. AB - Patients with chronic kidney disease (CKD) have a higher burden of atherosclerotic coronary artery disease compared with age- and gender-matched individuals with normal renal function. Cardiovascular calcification (CVC), a marker of atherosclerosis, is also more prevalent in these patients and is associated with serious clinical consequences. The pathogenesis of CVC is complex and includes factors that promote calcification and others that inhibit calcification. Thus, multiple therapeutic interventions should be used simultaneously to reduce the burden of calcification in patients with CKD. Thus far, interventional attempts have focused on curtailing the effects of factors that promote calcification such as management of known traditional factors for atherosclerotic coronary artery disease and on adopting specific approaches to normalize mineral metabolism, deliver adequate dialysis, and control serum cholesterol level. By contrast, interventions that may bolster the effects of inhibitors of calcification have not yet been studied well but are beginning to attract attention. Ideally, the goal of interventions is not only to slow or halt progression of calcification but also to reverse pre-existing calcification. Whether this goal is achievable is not currently known. This review examines the potential of various therapeutic interventions in reducing the CVC burden in patients with CKD. Moreover, the review is intended to stimulate more research in this area because the efficacy of these interventions has not been examined in controlled clinical trials. PMID- 16251251 TI - System B0,+ amino acid transport regulates the penetration stage of blastocyst implantation with possible long-term developmental consequences through adulthood. AB - Amino acid transport system B(0,+) was first characterized in detail in mouse blastocysts over two decades ago. Since then, this system has been shown to be involved in a wide array of developmental processes from blastocyst implantation in the uterus to adult obesity. Leucine uptake through system B(0,+) in blastocysts triggers mammalian target of rapamycin (mTOR) signalling. This signalling pathway selectively regulates development of trophoblast motility and the onset of the penetration stage of blastocyst implantation about 20 h later. Meanwhile, system B(0,+) becomes inactive in blastocysts a few hours before implantation in vivo. System B(0,+) can, however, be activated in preimplantation blastocysts by physical stimuli. The onset of trophoblast motility should provide the physiological physical stimulus activating system B(0,+) in blastocysts in vivo. Activation of system B(0,+) when trophoblast cells begin to penetrate the uterine epithelium would cause it to accumulate its preferred substrates, which include tryptophan, from uterine secretions. A low tryptophan concentration in external secretions next to trophoblast cells inhibits T-cell proliferation and rejection of the conceptus. Suboptimal system B(0,+) regulation of these developmental processes likely influences placentation and subsequent embryo nutrition, birth weight and risk of developing metabolic syndrome and obesity. PMID- 16251252 TI - Acyl-coenzyme a formation of simvastatin in mouse liver preparations. AB - Formation of an acyl-CoA thioester has been proposed, but not directly demonstrated, to be a key step in mediating both lactonization and atypical beta oxidation of 3-hydroxy-3-methylglutaryl-CoA reductase inhibitors. Here, we describe studies to characterize formation of acyl-CoA thioesters in vitro in mouse liver preparations using the hydroxy acid form of simvastatin (SVA) as a model substrate. With an optimized chromatography method, three new products were detected in addition to the dehydration product (P1) and the lactone form of simvastatin, which have been characterized previously (Prueksaritanont et al., 2001). Based on high-pressure liquid chromatography analysis, UV spectroscopy, mass spectrometry, and NMR spectral characterization, two metabolites were identified as acyl-CoA thioester conjugates of SVA and P1, respectively, whereas the third metabolite (M1) was confirmed to be the L-beta-hydroxy isomer of simvastatin. M1 was probably formed by stereospecific hydration, a previously reported reaction, and subsequent lactonization of P1-S-acyl CoA. Among all the mouse liver subcellular fractions, microsomes exhibited the highest capacity to catalyze the CoASH-dependent metabolism of SVA, whereas such activity was totally absent in cytosol. Together, these results provide direct experimental evidence that SVA (and conceivably other statins as well) is able to form an acyl-CoA thioester, possibly by microsomal long-chain acyl-CoA synthetase(s), leading to formation of two parallel metabolic pathways, one resulting in the two diastereomers of statin lactones (simvastatin and M1) and the other to the beta oxidation pathway of statin hydroxy acids. PMID- 16251254 TI - Metabolism of mometasone furoate and biological activity of the metabolites. AB - To better evaluate the pharmacokinetic and pharmacodynamic properties of the new inhaled glucocorticoid mometasone furoate (MF), the metabolism of MF was evaluated in rat and human tissues and in rat after i.v. administration. Metabolic studies with 3H-MF in human and rat plasma and S9 fractions of human and rat lung showed relatively high stability and a degradation pattern similar to that seen in buffer systems. MF was efficiently metabolized into at least five metabolites in S9 fractions of both rat and human liver. There were, however, quantitative differences in the metabolites between the two species. The apparent half-life of MF in the S9 fraction of human liver was found to be 3 times greater compared with that in rat. MET1, the most polar metabolite, was the major metabolite in rat liver fractions, whereas both MET1 and MET2 were formed to an equal extent in human liver. Metabolism and distribution studies in rats after intravenous and intratracheal administration of [1,2-(3)H]MF revealed that most of the radioactivity (approximately 90%) was present in the stomach, intestines, and intestinal contents, suggesting biliary excretion of MF and its metabolites. Radiochromatography showed that most radioactivity was associated with MET1, MET2, and MET 3. Fractionation of the high-performance liquid chromatography eluate (MET1-5) revealed that only MF [relative binding affinity (RBA) 2900] and MET2 (RBA 700) had appreciable glucocorticoid receptor binding affinity. These results suggest that MF undergoes distinct extrahepatic metabolism but generates active metabolites that might be in part responsible for the systemic side effects of MF. PMID- 16251253 TI - Nc100668, a new tracer for imaging of venous thromboembolism: disposition and metabolism in rats. AB - The 99mTc-complex of NC100668 [Acetyl-Asn-Gln-Glu-Gln-Val-Ser-Pro-Tyr(3-iodo)-Thr Leu-Leu-Lys-Gly-NC100194] is being evaluated for nuclear medical imaging of venous thromboembolism. NC100668 is a 13-amino acid peptide with a Tc-binding chelator [NC100194; -NH-CH2-CH2-N(CH2-CH2-NH-C(CH3)2-C(CH3)=N-OH)2] linked to the C-terminal end. Following injection in rats of [Asn-U-14C]NC100668 (labeling of the N-terminal amino acid), approximately 70% of the radioactivity was recovered in urine within 3 days. Following injection of [Lys-U-14C]NC100668 (labeling close to the C-terminal amino acid), radioactivity was cleared more slowly, with only 8% recovered in urine and approximately 80% of the radioactivity present in the body after 3 days. The highest concentration of radioactivity in the body following injection of [Lys-U-14C]NC100668 was observed in the kidney inner cortex; this probably represents 14C-labeled Lys, which is reabsorbed in the kidney tubules and incorporated into protein metabolism. Metabolite profiling by high-performance liquid chromatography with radiochemical detection revealed that following injection of [Asn-U-14C]NC100668, there is a rapid appearance in blood of one peak containing radioactive metabolite(s) originating from the N-terminal part of the molecule. In urine samples, only this radioactive peak was observed with no intact NC100668 remaining; this very hydrophilic N-terminal metabolite was probably either the N-terminal amino acid or a very short peptide. Liquid chromatography-mass spectrometry analyses of rat urine samples obtained after injection of nonlabeled NC100668 confirmed the identity of two metabolites generated from the C-terminal end of the molecule; Gly-NC100194 was identified as the major of these metabolites and NC100194 as a minor metabolite present at approximately one-tenth the amount of Gly-NC100194. No other metabolites were identified. PMID- 16251255 TI - Evidence for the bioactivation of zomepirac and tolmetin by an oxidative pathway: identification of glutathione adducts in vitro in human liver microsomes and in vivo in rats. AB - Although zomepirac (ZP) and tolmetin (TM) induce anaphylactic reactions and form reactive acyl glucuronides, a direct link between the two events remains obscure. We report herein that, in addition to acyl glucuronidation, both drugs are subject to oxidative bioactivation. Following incubations of ZP with human liver microsomes fortified with NADPH and glutathione (GSH), a metabolite with an MH+ ion at m/z 597 was detected by LC/MS/MS. On the basis of collision-induced dissociation and NMR evidence, the structure of this metabolite was determined to be 5-[4'-chlorobenzoyl]-1,4-dimethyl-3-glutathionylpyrrole-2-acetic acid (ZP-SG), suggesting that the pyrrole moiety of ZP had undergone oxidation to an epoxide intermediate, followed by addition of GSH and loss of the elements of H2O to yield the observed conjugate. The oxidative bioactivation of ZP most likely is catalyzed by cytochrome P450 (P450) 3A4, since the formation of ZP-SG was reduced to approximately 10% of control values following pretreatment of human liver microsomes with ketoconazole or with an inhibitory anti-P450 3A4 IgG. A similar GSH adduct, namely 5-[4'-methylbenzoyl]-1-methyl-3-glutathionylpyrrole-2-acetic acid (TM-SG), was identified when TM was incubated with human liver microsomal preparations. The relevance of these in vitro findings to the in vivo situation was established through the detection of the same thiol adducts in rats treated with ZP and TM, respectively. Taken together, these data suggest that, in addition to the formation of acyl glucuronides, oxidative metabolism of ZP and TM affords reactive species that may haptenize proteins and thereby contribute to the drug-mediated anaphylactic reactions. PMID- 16251256 TI - Real epidemiologists don't do ecological studies? PMID- 16251257 TI - Long-term mortality amongst Gulf War Veterans: is there a relationship with experiences during deployment and subsequent morbidity? AB - BACKGROUND: Gulf War Veterans have previously been shown to have, in the short term, an excess risk of death from 'external' (i.e. non-disease) causes of death. This study aims to determine whether there remains an excess of non-disease related deaths in Gulf Veterans, 13 years after deployment, and, for the first time, to determine whether there is a relationship between experiences reported in the Gulf, post-war symptoms, and subsequent mortality experience. METHODS: We conducted a cohort study with follow-up from April 1, 1991 (the end of the Gulf War) to June 30, 2004. Participants were 53 462 Gulf War Veterans and a cohort of military personnel, matched for age-group, sex, rank, service and level of fitness, who were not deployed to the Gulf. The outcome measure used was mortality as recorded on the NHS central register. RESULTS: There is no difference, 13 years after the end of the Gulf War, in the overall mortality experience of Gulf War Veterans. The excess in non-disease-related deaths previously reported is confined to the initial 7 years of follow-up [mortality rate ratio (MRR) 1.31, 95% confidence interval (CI) 1.06-1.63] rather than the more recent period (MRR 1.05, 95% CI 0.83-1.33). Overall experiences reported during Gulf deployment did not influence subsequent risk of dying, but there was non-significant increased risk of dying from a disease-related death (MRR 1.99, 95% CI 0.98-4.04) associated with reported exposure to depleted uranium and of a non-disease-related death associated with reporting handling of pesticides (MRR 2.05, 95% CI 0.91-4.61). Reporting of morbidity in the health surveys conducted was not related to future risk of death. CONCLUSION: The higher rates of non disease-related deaths in Gulf War Veterans is not evident in the period of follow-up since 1997. Neither the excess morbidity reported in health surveys nor the experiences during deployment significantly influenced future mortality. The two non-significant associations found (reported depleted uranium exposure and disease death, reporting handling pesticides and non-disease deaths) need to be considered in the context of the number of possible associations examined and potential biases-although they are biologically plausible. PMID- 16251258 TI - NSF for CHD: 3 years of 12-month follow-up audit after cardiac rehabilitation. AB - BACKGROUND: The coronary heart disease (CHD) National Service Framework (NSF) sets standards and milestones. For acute myocardial infarction (AMI) or coronary revascularization, 'Milestone 3, of Standard 12 requires that, by April 2002, every hospital should have clinical audit data no more than 12 months old showing 'total number and % of those recruited to cardiac rehabilitation who, one year after discharge, report: regular physical activity of at least 30 minutes duration on average five times a week, not smoking and a Body Mass Index (BMI) of <30 kg/m2'. This study looked at cost, method and practicalities of retrieving this data. METHODS: A postal questionnaire was used to follow-up coronary patients who started our cardiac rehabilitation programme between 1 April 2001 and 31 March 2004. The project was costed. RESULTS: Three hundred and seventy five (33 per cent) AMI patients, 412 (36 per cent) coronary artery bypass grafting (CABG) patients and 343 (30 per cent) percutaneous coronary intervention (PCI) patients entered the cardiac rehabilitation programme over 3 years. Completed questionnaires were received from 903 (80 per cent). Post-AMI patients or those stratified as high risk for further cardiac events were least likely to respond. Of responders, 74 per cent were exercising regularly, 95 per cent were not smoking and 79 per cent had a BMI <30 kg/m2. CONCLUSION: Targets for smoking and BMI set by the NSF are too low and were achieved by most patients before the start of cardiac rehabilitation. Patients who are post-AMI or are stratified as high risk need to be targeted if a high level of follow-up is to be achieved. PMID- 16251259 TI - Anticipatory activity of motor cortex in relation to rhythmic whisking. AB - Rats characteristically generate stereotyped exploratory (5-12 Hz) whisker movements, which can also be adaptively modulated. Here we tested the hypothesis that the vibrissal representation in motor cortex (vMCx) initiates and modulates whisking by acting on a subcortical whisking central pattern generator (CPG). We recorded local field potentials (LFPs) in vMCx of behaving Sprague-Dawley rats while monitoring whisking behavior through mystacial electromyograms (EMGs). Recordings were made during free exploration, under body restraint, or in a head fixed animal. LFP activity increased significantly prior to the onset of a whisking epoch and ended prior to the epoch's termination. In addition, shifts in whisking kinematics within a whisk epoch were often reflected in changes in LFP activity. These data support the hypothesis that vMCx may initiate and modulate whisking behavior through its action on a subcortical CPG. PMID- 16251260 TI - Olfactory cortical adaptation facilitates detection of odors against background. AB - Detection and discrimination of odors generally, if not always, occurs against an odorous background. On any given inhalation, olfactory receptor neurons will be activated by features of both the target odorant and features of background stimuli. To identify a target odorant against a background therefore, the olfactory system must be capable of grouping a subset of features into an odor object distinct from the background. Our previous work has suggested that rapid homosynaptic depression of afferents to the anterior piriform cortex (aPCX) contributes to both cortical odor adaptation to prolonged stimulation and habituation of simple odor-evoked behaviors. We hypothesize here that this process may also contribute to figure-ground separation of a target odorant from background stimulation. Single-unit recordings were made from both mitral/tufted cells and aPCX neurons in urethan-anesthetized rats and mice. Single-unit responses to odorant stimuli and their binary mixtures were determined. One of the odorants was randomly selected as the background and presented for 50 s. Forty seconds after the onset of the background stimulus, the second target odorant was presented, producing a binary mixture. The results suggest that mitral/tufted cells continue to respond to the background odorant and, when the target odorant is presented, had response magnitudes similar to that evoked by the binary mixture. In contrast, aPCX neurons filter out the background stimulus while maintaining responses to the target stimulus. Thus the aPCX acts as a filter driven most strongly by changing stimuli, providing a potential mechanism for olfactory figure-ground separation and selective reading of olfactory bulb output. PMID- 16251261 TI - Alpha-adrenoceptive dual modulation of inhibitory GABAergic inputs to Purkinje cells in the mouse cerebellum. AB - Noradrenaline (NA) modulates synaptic transmission in various sites of the CNS. In the cerebellar cortex, several studies have revealed that NA enhances inhibitory synaptic transmission by beta-adrenoceptor-and cyclic AMP-dependent pathways. However, the effects of alpha-adrenoceptor activation on cerebellar inhibitory neurotransmission have not yet been fully elucidated. Therefore we investigated the effects of the alpha1- or alpha2-adrenoceptor agonist on inhibitory postsynaptic currents (IPSCs) recorded from mouse Purkinje cells (PCs). We found that the nonselective alpha-adrenoceptor agonist 6-fluoro norepinephrine increased both the frequency and amplitude of spontaneous IPSCs (sIPSCs). This enhancement was mostly mimicked by the selective alpha1 adrenoceptor agonist phenylephrine (PE). PE also enhanced the amplitude of evoked IPSCs (eIPSCs) and increased the frequency but not the amplitude of miniature IPSCs (mIPSCs). Moreover, PE decreased the paired-pulse ratio of eIPSCs and did not change gamma-aminobutyric acid (GABA) receptor sensitivity in PCs. Conversely, the selective alpha2-adrenoceptor agonist clonidine significantly reduced both the frequency and the amplitude of sIPSCs. Neither eIPSCs nor mIPSCs were affected by clonidine. Furthermore, presynaptic cell-attached recordings showed that spontaneous activity of GABAergic interneurons was enhanced by PE but reduced by clonidine. These results suggest that NA enhances inhibitory neurotransmitter release by alpha1-adrenoceptors, which are expressed in presynaptic terminals and somatodendritic domains, whereas NA suppresses the excitability of interneurons by alpha2-adrenoceptors, which are expressed in presynaptic somatodendritic domains. Thus cerebellar alpha-adrenoceptors play roles in a presynaptic dual modulation of GABAergic inputs from interneurons to PCs, thereby providing a likely mechanism for the fine-tuning of information flow in the cerebellar cortex. PMID- 16251262 TI - Primate upper limb muscles exhibit activity patterns that differ from their anatomical action during a postural task. AB - The present study examined muscular activity in the primate proximal forelimb during a posture task. By applying loads selectively to the shoulder, elbow, or both joints, we observed that monoarticular shoulder and elbow muscles varied their activity with loads at the unspanned joint. Shoulder monoarticulars changed activity with elbow torque and elbow monoarticulars changed activity with shoulder torque. Due to this additional modulation, the maximal activation of monoarticular muscles was deviated from their anatomical action toward either shoulder-extension/elbow-flexion or shoulder-flexion/elbow-extension. Biarticular muscles also expressed deviations in their preferred torque direction toward either shoulder-extension/elbow-flexion or shoulder-flexion/elbow-extension. The biased distribution of preferred torque directions in proximal forelimb muscles could be modeled by the minimization of a global measure of muscle activity. Moreover, arm-related neurons of primary motor cortex exhibit a similar bias in preferred torque directions consistent with the intimate relationship between the primary motor cortex and the motor periphery. PMID- 16251263 TI - Corticospinal excitability is lower during rhythmic arm movement than during tonic contraction. AB - Humans perform rhythmic, locomotor movements with the arms and legs every day. Studies using reflexes to probe the functional role of the CNS suggest that spinal circuits are an important part of the neural control system for rhythmic arm cycling and walking. Here, by studying motor-evoked potentials (MEPs) in response to transcranial magnetic stimulation (TMS) of the motor cortex, and H reflexes induced by electrical stimulation of peripheral nerves, we show a reduction in corticospinal excitability during rhythmic arm movement compared with tonic, voluntary contraction. Responses were compared between arm cycling and tonic contraction at four positions, while participants generated similar levels of muscle activity. Both H-reflexes and MEPs were significantly smaller during arm cycling than during tonic contraction at the midpoint of arm flexion (F = 13.51, P = 0.006; F = 11.83, P = 0.009). Subthreshold TMS significantly facilitated the FCR H-reflex during tonic contractions, but did not significantly modulate H-reflex amplitude during arm cycling. The data indicate a reduction in the responsiveness of cells constituting the fast, monosynaptic, corticospinal pathway during arm cycling and suggest that the motor cortex may contribute less to motor drive during rhythmic arm movement than during tonic, voluntary contraction. Our results are consistent with the idea that subcortical regions contribute to the control of rhythmic arm movements despite highly developed corticospinal projections to the human upper limb. PMID- 16251264 TI - Excitatory GABAergic effects in striatal projection neurons. AB - The ability of synaptically released GABA to facilitate action potential generation in striatal projection neurons was studied in brain slices using current-clamp, gramicidin-perforated whole cell recordings. Evoked GABAergic postsynaptic potentials (PSPs) were pharmacologically isolated with ionotropic glutamate receptor antagonists. Subthreshold depolarizing current injections were paired with GABAergic PSPs at different intervals. GABAergic PSPs were able to convert current injection-induced depolarizations from subthreshold to suprathreshold, but only when they preceded the current injection by an appropriate interval; accordingly, action potentials were observed 4-140 ms after the onset of the GABAergic PSP, and their likelihood was maximal after 50-60 ms. The GABAergic excitatory effects were fully blocked by the GABA(A) receptor antagonist bicuculline. Appropriately timed GABA PSPs decreased the time taken by current injections to depolarize projection neurons, causing an apparent reduction in the spike threshold. In control solution, the ability of evoked PSPs (comprising both glutamatergic and GABAergic components) to reach spike threshold was often impaired by bicuculline. We conclude that GABAergic PSPs can exert excitatory effects on projection neurons and that this ability crucially depends on the timing between the GABAergic event and a concomitant depolarizing input. PMID- 16251265 TI - Functional properties of grasping-related neurons in the ventral premotor area F5 of the macaque monkey. AB - We investigated the motor and visual properties of F5 grasping neurons, using a controlled paradigm that allows the study of the neuronal discharge during both observation and grasping of many different three-dimensional objects with and without visual guidance. All neurons displayed a preference for grasping of an object or a set of objects. The same preference was maintained when grasping was performed in the dark without visual feedback. In addition to the motor-related discharge, about half of the neurons also responded to the presentation of an object or a set of objects, even when a grasping movement was not required. Often the object evoking the strongest activity during grasping also evoked optimal activity during its visual presentation. Hierarchical cluster analysis indicated that the selectivity of both the motor and the visual discharge of the F5 neurons is determined not by the object shape but by the grip posture used to grasp the object. Because the same paradigm has been used to study the properties of hand grasping neurons in the dorsal premotor area F2, and in the anterior intraparietal area (AIP), a comparison of the functional properties of grasping related neurons in the three cortical areas (F5, F2, AIP) is addressed for the first time. PMID- 16251266 TI - Central representation of dynamics when manipulating handheld objects. AB - To explore the neural mechanisms related to representation of the manipulation dynamics of objects, we performed whole-brain fMRI while subjects balanced an object in stable and highly unstable states and while they balanced a rigid object and a flexible object in the same unstable state, in all cases without vision. In this way, we varied the extent to which an internal model of the manipulation dynamics was required in the moment-to-moment control of the object's orientation. We hypothesized that activity in primary motor cortex would reflect the amount of muscle activation under each condition. In contrast, we hypothesized that cerebellar activity would be more strongly related to the stability and complexity of the manipulation dynamics because the cerebellum has been implicated in internal model-based control. As hypothesized, the dynamics related activation of the cerebellum was quite different from that of the primary motor cortex. Changes in cerebellar activity were much greater than would have been predicted from differences in muscle activation when the stability and complexity of the manipulation dynamics were contrasted. On the other hand, the activity of the primary motor cortex more closely resembled the mean motor output necessary to execute the task. We also discovered a small region near the anterior edge of the ipsilateral (right) inferior parietal lobule where activity was modulated with the complexity of the manipulation dynamics. We suggest that this is related to imagining the location and motion of an object with complex manipulation dynamics. PMID- 16251267 TI - Effect of benzodiazepine hypnotic triazolam on relationship of blood pressure and Paco2 to cerebral blood flow during human non-rapid eye movement sleep. AB - We sought to clarify the effect of short-acting benzodiazepine hypnotic on the relationship of arterial blood pressure and arterial partial pressure of carbon dioxide (Paco2) to regional cerebral blood flow (rCBF) during human non-rapid-eye movement (non-REM) sleep. Nine young normal volunteers were treated in a randomized, crossover design with triazolam or placebo and underwent positron emission tomography at night. During wakefulness and stage 2 and slow wave (stages 3 and 4) sleep, we measured mean arterial blood pressure (MAP), Paco2, and absolute CBF. With triazolam compared to placebo, MAP reduced gradually. During stage 2 sleep, Paco2 increased and whole-brain mean CBF decreased. With triazolam, relative rCBF of the left orbital basal forebrain decreased more during stage 2 than slow wave sleep, whereas absolute CBF of the occipital cortex and cerebral white matter remained constant. During triazolam-induced stage 2 sleep, absolute CBF of the cerebral white matter correlated more strongly to both MAP and Paco2 than during placebo sleep and also correlated more strongly to both MAP and Paco2 than absolute CBF of the occipital cortex. In the frontal white matter, during triazolam-induced stage 2 sleep compared to wakefulness, absolute CBF was significantly better correlated to MAP, but not to Paco2. During triazolam-induced stage 2, the cerebral white matter may receive a modulated CBF regulation having the strengthened relationship of Paco2 to CBF and, more locally, the frontal white matter may depend precariously on CBF regulation. PMID- 16251268 TI - Building native protein conformation from highly approximate backbone torsion angles. AB - Reconstructing a protein in three dimensions from its backbone torsion angles is an ongoing challenge because minor inaccuracies in these angles produce major errors in the structure. As a familiar example, a small change in an elbow angle causes a large displacement at the end of your arm, the longer the arm, the larger the displacement. Even accurate knowledge of the backbone torsions and Psi is insufficient, owing to the small, but cumulative, deviations from ideality in backbone planarity, which, if ignored, also lead to major errors in the structure. Against this background, we conducted a computational experiment to assess whether protein conformation can be determined from highly approximate backbone torsion angles, the kind of information that is now obtained readily from NMR. Specifically, backbone torsion angles were taken from proteins of known structure and mapped into 60 degrees x 60 degrees grid squares, called mesostates. Side-chain atoms beyond the beta -carbon were discarded. A mesostate representation of the protein backbone was then used to extract likely candidates from a fragment library of mesostate pentamers, followed by Monte Carlo-based fragment-assembly simulations to identify stable conformations compatible with the given mesostate sequence. Only three simple energy terms were used to gauge stability: molecular compaction, soft-sphere repulsion, and hydrogen bonding. For the six representative proteins described here, stable conformers can be partitioned into a remarkably small number of topologically distinct clusters. Among these, the native topology is found with high frequency and can be identified as the cluster with the most favorable energy. PMID- 16251269 TI - A model of anthrax toxin lethal factor bound to protective antigen. AB - Anthrax toxin is made up of three proteins: the edema factor (EF), lethal factor (LF) enzymes, and the multifunctional protective antigen (PA). Proteolytically activated PA heptamerizes, binds the EF/LF enzymes, and forms a pore that allows for EF/LF passage into host cells. Using directed mutagenesis, we identified three LF-PA contact points defined by a specific disulfide crosslink and two pairs of complementary charge-reversal mutations. These contact points were consistent with the lowest energy LF-PA complex found by using Rosetta protein protein docking. These results illustrate how biochemical and computational methods can be combined to produce reliable models of large complexes. The model shows that EF and LF bind through a highly electrostatic interface, with their flexible N-terminal region positioned at the entrance of the heptameric PA pore and thus poised to initiate translocation in an N- to C-terminal direction. PMID- 16251270 TI - Regulation of the Caenorhabditis elegans oxidative stress defense protein SKN-1 by glycogen synthase kinase-3. AB - Oxidative stress plays a central role in many human diseases and in aging. In Caenorhabditis elegans the SKN-1 protein induces phase II detoxification gene transcription, a conserved oxidative stress response, and is required for oxidative stress resistance and longevity. Oxidative stress induces SKN-1 to accumulate in intestinal nuclei, depending on p38 mitogen-activated protein kinase signaling. Here we show that, in the absence of stress, phosphorylation by glycogen synthase kinase-3 (GSK-3) prevents SKN-1 from accumulating in nuclei and functioning constitutively in the intestine. GSK-3 sites are conserved in mammalian SKN-1 orthologs, indicating that this level of regulation may be conserved. If inhibition by GSK-3 is blocked, background levels of p38 signaling are still required for SKN-1 function. WT and constitutively nuclear SKN-1 comparably rescue the skn-1 oxidative stress sensitivity, suggesting that an inducible phase II response may provide optimal stress protection. We conclude that (i) GSK-3 inhibits SKN-1 activity in the intestine, (ii) the phase II response integrates multiple regulatory signals, and (iii), by inhibiting this response, GSK-3 may influence redox conditions. PMID- 16251271 TI - Formation of apoptosome is initiated by cytochrome c-induced dATP hydrolysis and subsequent nucleotide exchange on Apaf-1. AB - Apoptosis in metazoans is executed by a group of intracellular proteases named caspases. One of the caspase-activating pathways in mammals is initiated by the release of cytochrome c from mitochondria to cytosol, where it binds to Apaf-1 to form a procaspase-9-activating heptameric protein complex named apoptosome. We report here the reconstitution of this pathway with purified recombinant Apaf-1, procaspase-9, procaspase-3, and cytochrome c from horse heart. Apaf-1 contains a dATP as a cofactor. Cytochrome c binding to Apaf-1 induces hydrolysis of dATP to dADP, which is subsequently replaced by exogenous dATP. The dATP hydrolysis and exchange on Apaf-1 are two required steps for apoptosome formation. PMID- 16251273 TI - Mouse lacking COUP-TFII as an animal model of Bochdalek-type congenital diaphragmatic hernia. AB - Congenital diaphragmatic hernia (CDH), a life-threatening anomaly, is a major cause of pediatric mortality. Although the disease was described >350 years ago, the etiology of CDH is poorly understood. Here, we show that tissue-specific null mutants of COUP-TFII exhibit Bochdalek-type CDH, the most common form of CDH. COUP-TFII, a member of orphan nuclear receptors, is expressed in regions critical for the formation of the diaphragm during embryonic development. Ablation of COUP TFII in the foregut mesenchyme, including the posthepatic mesenchymal plate (PHMP), results in the malformation of the diaphragm and the failure of appropriate attachment of the PHMP to the body wall. Thus, both the stomach and liver enter the thoracic cavity, leading to lung hypoplasia and neonatal death. Recently a minimally deleted region for CDH has been identified on chromosome 15q26.1-26.2 by CGH array and FISH analysis. COUP-TFII is one of the four known genes residing within this critical region. Our finding suggests that COUP-TFII is a likely contributor to the formation of CDH in individuals with 15q deletions, and it may also be a potential contributor to some other Bochdalek type of CDH. PMID- 16251274 TI - Adhesion controls bacterial actin polymerization-based movement. AB - As part of its infectious life cycle, the bacterial pathogen Listeria monocytogenes propels itself through the host-cell cytoplasm by triggering the polymerization of host-cell actin near the bacterial surface, harnessing the activity of several cytoskeletal proteins used during actin-based cell crawling. To distinguish among several classes of biophysical models of actin-based bacterial movement, we used a high-throughput tracking technique to record the movement of many individual bacteria during temperature shifts. The speed of each bacterium varied strongly with temperature, closely following the Arrhenius rate law. Among bacteria, the prefactor A of the Arrhenius dependence unexpectedly varied exponentially with apparent activation energy, E(a), over a wide range (8 21 kcal/mol), reminiscent of the "rate compensation effect" of classical catalytic reactions. Average E(a) were increased for mutant bacteria deficient in binding Ena/VASP proteins and bacteria moving in diluted extract. These two effects were additive. The observed temperature and rate compensation effects are consistent with a class of simple kinetic models in which the bacterium advances through the thermally driven, cooperative breakage of groups of adhesive bonds on its surface. The estimated number of coupled adhesive bonds N on the bacterial surface varies between 10 and 40 bonds. In contrast to other models, this model correctly predicts an experimentally observed negative correlation between bacterial speed and actin gel density. The idea that speed depends on adhesion, rather than polymerization, suggests several alternative mechanisms by which known cytoskeletal regulatory proteins could control cellular movement. PMID- 16251272 TI - Regulation of RNA splicing by the methylation-dependent transcriptional repressor methyl-CpG binding protein 2. AB - Rett syndrome (RTT) is a postnatal neurodevelopmental disorder characterized by the loss of acquired motor and language skills, autistic features, and unusual stereotyped movements. RTT is caused by mutations in the X-linked gene encoding methyl-CpG binding protein 2 (MeCP2). Mutations in MECP2 cause a variety of neurodevelopmental disorders including X-linked mental retardation, psychiatric disorders, and some cases of autism. Although MeCP2 was identified as a methylation-dependent transcriptional repressor, transcriptional profiling of RNAs from mice lacking MeCP2 did not reveal significant gene expression changes, suggesting that MeCP2 does not simply function as a global repressor. Changes in expression of a few genes have been observed, but these alterations do not explain the full spectrum of Rett-like phenotypes, raising the possibility that additional MeCP2 functions play a role in pathogenesis. In this study, we show that MeCP2 interacts with the RNA-binding protein Y box-binding protein 1 and regulates splicing of reporter minigenes. Importantly, we found aberrant alternative splicing patterns in a mouse model of RTT. Thus, we uncovered a previously uncharacterized function of MeCP2 that involves regulation of splicing, in addition to its role as a transcriptional repressor. PMID- 16251275 TI - Integration and evolution of the cichlid mandible: the molecular basis of alternate feeding strategies. AB - African cichlid fishes have repeatedly evolved highly specialized modes of feeding through adaptations of their oral jaws. Here, we explore the molecular genetic basis of the opening and closing lever mechanisms of the cichlid lower jaw, which have traditionally been used to describe the mechanics of feeding behavior in bony fishes. Quantitative genetic analyses demonstrate that the opening and closing mechanisms are genetically modular and therefore free to evolve independently. Bmp4 (bone morphogenetic protein 4) is one of two loci that segregate with the mechanical advantage of closing and that together account for >30% of the phenotypic variance in this trait. Species-specific differences in jaw shape are obvious early in cichlid larval development and are correlated with patterns of bmp4 expression in the mandibular primordium. When bmp4 is overexpressed in the obligate suction feeder Danio rerio, mandibular morphology exhibits specific transformations of opening and closing lever ratios. We conclude that patterns of morphological integration of the cichlid jaw reflect a balance among conflicting functional demands. Further, we demonstrate that bmp4 has the potential to alter mandibular morphology in a way that mimics adaptive variation among fish species. PMID- 16251276 TI - Mutagenesis-based definitions and probes of residue burial in proteins. AB - Every residue of the 101-aa Escherichia coli toxin CcdB was substituted with Ala, Asp, Glu, Lys, and Arg by using site-directed mutagenesis. The activity of each mutant in vivo was characterized as a function of Controller of Cell Division or Death B protein (CcdB) transcriptional level. The mutation data suggest that an accessibility value of 5% is an appropriate cutoff for definition of buried residues. At all buried positions, introduction of Asp results in an inactive phenotype at all CcdB transcriptional levels. The average amount of destabilization upon substitution at buried positions decreases in the order Asp>Glu>Lys>Arg>Ala. Asp substitutions at buried sites in two other proteins, maltose-binding protein and thioredoxin, also were shown to be severely destabilizing. Ala and Asp scanning mutagenesis, in combination with dose dependent expression phenotypes, was shown to yield important information on protein structure and activity. These results also suggest that such scanning mutagenesis data can be used to rank order sequence alignments and their corresponding homology models, as well as to distinguish between correct and incorrect structural alignments. With continuous reductions in oligonucleotide costs and increasingly efficient site-directed mutagenesis procedures, comprehensive scanning mutagenesis experiments for small proteins/domains are quite feasible. PMID- 16251277 TI - Dual targeting is the rule for organellar aminoacyl-tRNA synthetases in Arabidopsis thaliana. AB - In plants, protein synthesis occurs in the cytosol, mitochondria, and plastids. Each compartment requires a full set of tRNAs and aminoacyl-tRNA synthetases. We have undertaken a systematic analysis of the targeting of organellar aminoacyl tRNA synthetases in the model plant Arabidopsis thaliana. Dual targeting appeared to be a general rule. Among the 24 identified organellar aminoacyl-tRNA synthetases (aaRSs), 15 (and probably 17) are shared between mitochondria and plastids, and 5 are shared between cytosol and mitochondria (one of these aaRSs being present also in chloroplasts). Only two were shown to be uniquely chloroplastic and none to be uniquely mitochondrial. Moreover, there are no examples where the three aaRS genes originating from the three ancestral genomes still coexist. These results indicate that extensive exchange of aaRSs has occurred during evolution and that many are now shared between two or even three compartments. The findings have important implications for studies of the translation machinery in plants and on protein targeting and gene transfer in general. PMID- 16251279 TI - Population pharmacokinetics of rifapentine and its primary desacetyl metabolite in South African tuberculosis patients. AB - This study was designed to describe the population pharmacokinetics of rifapentine (RFP) and 25-desacetyl RFP in a South African pulmonary tuberculosis patient population. Special reference was made to studying the influence of previous exposure to rifampin (RIF) and the variability in pharmacokinetic parameters between patients and between occasions and the influence of different covariates. Patients were included in the study if they had been receiving first line antimycobacterial therapy (rifampin, isoniazid, pyrazinamide, and ethambutol) for not less than 4 weeks and not more than 6 weeks and were divided into three RFP dosage groups based on weight: 600 mg, <45 kg; 750 mg, 46 to 55 kg; and 900 mg, >55 kg. Participants received a single oral dose of RFP together with concomitant antimycobacterial agents, excluding RIF, on study days 1 and 5 after they ingested a soup-based meal. The RFP and 25-desacetyl RFP concentration time data were analyzed by nonlinear mixed-effect modeling using NONMEM. The pharmacokinetics of the parent drug were modeled separately, and the individual pharmacokinetic parameters were used as inputs for the 25-desacetyl RFP pharmacokinetic model. A one-compartment disposition model was found to best describe the data for both the parent and the metabolite, and the metabolite was assumed to be formed only from the central compartment of the parent drug. Prior treatment with RIF did not alter the pharmacokinetics of RFP but appeared to increase the excretion of 25-desacetyl RFP in a nonlinear fashion. The RFP oral clearance and volume of distribution were found to increase by 0.049 liter/h and 0.691 liter, respectively, with a 1-kg increase from the median weight of 50 kg. The oral clearance of 25-desacetyl RFP was found to be 35% lower in female patients. The model developed here describes the population pharmacokinetics of RFP and its primary metabolite in tuberculosis patients and includes the effects of prior administration with RIF and covariate factors. PMID- 16251278 TI - A genetic approach to access serotonin neurons for in vivo and in vitro studies. AB - Serotonin (5HT) is a critical modulator of neural circuits that support diverse behaviors and physiological processes, and multiple lines of evidence implicate abnormal serotonergic signaling in psychiatric pathogenesis. The significance of 5HT underscores the importance of elucidating the molecular pathways involved in serotonergic system development, function, and plasticity. However, these mechanisms remain poorly defined, owing largely to the difficulty of accessing 5HT neurons for experimental manipulation. To address this methodological deficiency, we present a transgenic route to selectively alter 5HT neuron gene expression. This approach is based on the ability of a Pet-1 enhancer region to direct reliable 5HT neuron-specific transgene expression in the CNS. Its versatility is illustrated with several transgenic mouse lines, each of which provides a tool for 5HT neuron studies. Two lines allow Cre-mediated recombination at different stages of 5HT neuron development. A third line in which 5HT neurons are marked with yellow fluorescent protein will have numerous applications, including their electrophysiological characterization. To demonstrate this application, we have characterized active and passive membrane properties of midbrain reticular 5HT neurons, which heretofore have not been reported to our knowledge. A fourth line in which Pet-1 loss of function is rescued by expression of a Pet-1 transgene demonstrates biologically relevant levels of transgene expression and offers a route for investigating serotonergic protein structure and function in a behaving animal. These findings establish a straightforward and reliable approach for developing an array of tools for in vivo and in vitro studies of 5HT neurons. PMID- 16251280 TI - Application of quantitative real-time reverse transcription-PCR in assessing drug efficacy against the intracellular pathogen Cryptosporidium parvum in vitro. AB - We report here on a quantitative real-time reverse transcription-PCR (qRT-PCR) assay for assessing drug efficacy against the intracellular pathogen Cryptosporidium parvum. The qRT-PCR assay detects 18S rRNA transcripts from both parasites, that is, the cycle threshold for 18S rRNA from parasites (C(T)([P18S])) and host cells (C(T)([H18S])), and evaluates the relative expression between parasite and host rRNA levels (i.e., deltaC(T) = C(T)([P18S]) C(T)([H18S])) to minimize experimental and operational errors. The choice of qRT PCR over quantitative PCR (qPCR) in this study is based on the observations that (i) the relationship between the logarithm of infected parasites (log[P]) and the normalized relative level of rRNA (deltadeltaC(T)) is linear, with a fourfold dynamic range, by qRT-PCR but sigmoidal (nonlinear) by qPCR; and (ii) the level of RNA represents that of live parasites better than that of DNA, because the decay of RNA (99% in approximately 3 h) in dead parasites is faster than that of DNA (99% in approximately 24 to 48 h) under in vitro conditions. The reliability of the qRT-PCR method was validated by testing the efficacies of nitazoxanide and paromomycin on the development of two strains of C. parvum (IOWA and KSU-1) in HCT-8 cells in vitro. Both compounds displayed dose-dependent inhibitions. The observed MIC50 values for nitazoxanide and paromomycin were 0.30 to 0.45 micro/ml and 89.7 to 119.0 microg/ml, respectively, comparable to the values reported previously. Using the qRT-PCR assay, we have also observed that pyrazole could inhibit C. parvum development in vitro (MIC50 = 15.8 mM), suggesting that the recently discovered Cryptosporidium alcohol dehydrogenases may be explored as new drug targets. PMID- 16251281 TI - TEM-109 (CMT-5), a natural complex mutant of TEM-1 beta-lactamase combining the amino acid substitutions of TEM-6 and TEM-33 (IRT-5). AB - Escherichia coli CF349 exhibited a complex beta-lactam resistance phenotype, including resistance to amoxicillin and ticarcillin alone and in combination with clavulanate and to some extended-spectrum cephalosporins. The double-disk synergy test was positive. CF349 harbored an 85-kb conjugative plasmid which encoded a beta-lactamase of pI 5.9. The corresponding bla gene was identified by PCR and sequencing as a bla(TEM) gene. The deduced protein sequence revealed a new complex mutant of TEM-1 beta-lactamase designated TEM-109 (CMT-5). TEM-109 contained both the substitutions Glu104Lys and Arg164His of the expanded-spectrum beta-lactamase (ESBL) TEM-6 and Met69Leu of the inhibitor-resistant TEM-33 (IRT 5). TEM-109 exhibited hydrolytic activity against ceftazidime similar to that of TEM-6 (k(cat), 56 s(-1) and 105 s(-1), respectively; K(m) values, 226 and 247 microM, respectively). The 50% inhibitory concentrations of clavulanate and tazobactam (0.13 microM and 0.27 microM, respectively) were 5- to 10-fold higher for TEM-109 than for TEM-6 (0.01 and 0.06 microM, respectively) but were almost 10-fold lower than those for TEM-33. The characterization of this novel CMT, which exhibits a low level of resistance to inhibitors, highlights the emergence of this new ESBL type. PMID- 16251282 TI - Antibiotic abscess penetration: fosfomycin levels measured in pus and simulated concentration-time profiles. AB - The present study was performed to evaluate the ability of fosfomycin, a broad spectrum antibiotic, to penetrate into abscess fluid. Twelve patients scheduled for surgical or computer tomography-guided abscess drainage received a single intravenous dose of 8 g of fosfomycin. The fosfomycin concentrations in plasma over time and in pus upon drainage were determined. A pharmacokinetic model was developed to estimate the concentration-time profile of fosfomycin in pus. Individual fosfomycin concentrations in abscess fluid at drainage varied substantially, ranging from below the limit of detection up to 168 mg/liter. The fosfomycin concentrations in pus of the study population correlated neither with plasma levels nor with the individual ratios of abscess surface area to volume. This finding was attributed to highly variable abscess permeability. The average concentration in pus was calculated to be 182 +/- 64 mg/liter at steady state, exceeding the MIC(50/90)s of several bacterial species which are commonly involved in abscess formation, such as streptococci, staphylococci, and Escherichia coli. Hereby, the exceptionally long mean half-life of fosfomycin of 32 +/- 39 h in abscess fluid may favor its antimicrobial effect because fosfomycin exerts time-dependent killing. After an initial loading dose of 10 to 12 g, fosfomycin should be administered at doses of 8 g three times per day to reach sufficient concentrations in abscess fluid and plasma. Applying this dosing regimen, fosfomycin levels in abscess fluid are expected to be effective after multiple doses in most patients. PMID- 16251283 TI - Fitness cost due to mutations in the 16S rRNA associated with spectinomycin resistance in Chlamydia psittaci 6BC. AB - The fitness cost of a resistance determinant is the primary parameter that determines its frequency in vivo. As a model for analysis of the impact of drug resistance mutations on the intracellular life cycle of Chlamydia spp., we studied the growth of four genetically defined spectinomycin-resistant (Spc(r)) clonal variants of Chlamydia psittaci 6BC isolated in the plaque assay. The development of each variant was monitored over 46 h postinfection in the absence of drug, either in pure culture or in 1:1 competition with the parent strain. Spc(r) mutations in the 16S rRNA gene at positions 1191 and 1193 were associated with a marked impairment of C.psittaci biological fitness, and the bacteria were severely out-competed by the wild-type parent. In contrast, mutations at position 1192 had minor effects on the bacterial life cycle, allowing the resistant isolates to compete more efficiently with the wild-type strain. Thus, mutations with a wide range of fitness costs can be selected in the plaque assay, providing a new strategy for prediction and monitoring of the emergence of antibiotic resistance in chlamydiae. So far, drug resistance has not been a serious threat for the treatment of chlamydial infections. Tetracycline is an effective antichlamydial drug that targets 16S rRNA. Attempts to isolate spontaneous tetracycline-resistant mutants of C. psittaci 6BC revealed a frequency <3 x 10( 9). We suggest that the rarity of genotypic antibiotic resistance among chlamydial clinical isolates reflects the deleterious effects of such mutations on the fitness of these obligate intracellular bacteria in the host. PMID- 16251284 TI - Anti-human immunodeficiency virus type 1 activity of the nonnucleoside reverse transcriptase inhibitor GW678248 in combination with other antiretrovirals against clinical isolate viruses and in vitro selection for resistance. AB - GW678248, a novel nonnucleoside reverse transcriptase inhibitor, has been evaluated for anti-human immunodeficiency virus activity in a variety of in vitro assays against laboratory strains and clinical isolates. When GW678248 was tested in combination with approved drugs in the nucleoside and nucleotide reverse transcriptase inhibitor classes or the protease inhibitor class, the antiviral activities were either synergistic or additive. When GW678248 was tested in combination with approved drugs in the nonnucleoside reverse transcriptase inhibitor class, the antiviral activities were either additive or slightly antagonistic. Clinical isolates from antiretroviral drug-experienced patients were selected for evaluation of sensitivity to GW678248 in a recombinant virus assay. Efavirenz (EFV) and nevirapine (NVP) had > or = 10-fold increases in their 50% inhibitory concentrations (IC50s) for 85% and 98% of the 55 selected isolates, respectively, whereas GW678248 had a > or = 10-fold increase in the IC50 for only 17% of these isolates. Thus, 81 to 83% of the EFV- and/or NVP resistant viruses from this data set were susceptible to GW678248. Virus populations resistant to GW678248 were selected by in vitro dose-escalating serial passage. Resistant progeny viruses recovered after eight passages had amino acid substitutions V106I, E138K, and P236L in the reverse transcriptase coding region in one passage series and amino acid substitutions K102E, V106A, and P236L in a second passage series. PMID- 16251285 TI - Photodynamic sensitization of Leishmania amazonensis in both extracellular and intracellular stages with aluminum phthalocyanine chloride for photolysis in vitro. AB - Leishmania amazonensis, a causative agent of cutaneous leishmaniasis, is susceptible in vitro to light-mediated cytolysis in the presence of or after pretreatment with the photosensitizer aluminum phthalocyanine chloride. Cytolysis of both promastigotes and axenic amastigotes required less photosensitizer (e.g., one microg.ml(-1)) and a lower light dose (e.g., 1.5 J.cm(-2)) than did the mammalian cells examined for comparison. Exposure of Leishmania cells to the photosensitizer alone had little effect on their viability, as judged from their motility, growth, and/or retention of green fluorescent proteins genetically engineered for episomal expression. Fluorimetric assays for cell-associated and released green fluorescence proteins proved to be even more sensitive for the evaluation of cell viability than microscopy for the evaluation of motility and/or integrity. Axenic amastigotes pretreated with the photosensitizer infected macrophages of the J774 line but were lysed intracellularly when the infected cells were exposed to light. Addition of the photosensitizer to the already infected cells produced no effect on their intracellular parasites. However, light irradiation lysed these macrophages and also those infected with parasites preincubated with the photosensitizer at a concentration of 5 microg.ml(-1) or higher. Photosensitized Leishmania cells are highly susceptible to cytolysis, apparently due to the generation of reactive oxidative species on light illumination, suggestive of inefficiency of their antioxidant mechanisms. Efficient delivery of photosensitizers to intracellular Leishmania is expected to increase their therapeutic potentials against leishmaniasis. PMID- 16251286 TI - Novel acquired metallo-beta-lactamase gene, bla(SIM-1), in a class 1 integron from Acinetobacter baumannii clinical isolates from Korea. AB - Carbapenem resistance mediated by acquired carbapenemase genes has been increasingly reported, particularly for clinical isolates of Pseudomonas aeruginosa and Acinetobacter spp. Of 1,234 non-duplicate isolates of carbapenem resistant Pseudomonas spp. and Acinetobacter spp. isolated at a tertiary-care hospital in Seoul, Korea, 211 (17%) were positive for metallo-beta-lactamase (MBL). Of these, 204 (96%) had either the bla(IMP-1) or bla(VIM-2) allele. In addition, seven Acinetobacter baumannii isolates were found to have a novel MBL gene, which was designated bla(SIM-1). The SIM-1 protein has a pI of 7.2, is a new member of subclass B1, and exhibits 64 to 69% identity with the IMP-type MBLs, which are its closest relatives. All SIM-1-producing isolates exhibited relatively low imipenem and meropenem MICs (8 to 16 microg/ml) and had a multidrug resistance phenotype. Expression of the cloned bla(SIM-1) gene in Escherichia coli revealed that the encoded enzyme is capable of hydrolyzing a broad array of beta-lactams, including penicillins, narrow- to expanded-spectrum cephalosporins, and carbapenems. The bla(SIM-1) gene was carried on a gene cassette inserted into a class 1 integron, which included three additional cassettes (arr-3, catB3, and aadA1). The strains were isolated from sputum and urine specimens from patients with pneumonia and urinary tract infections, respectively. All patients had various underlying diseases. Pulsed-field gel electrophoresis of SmaI-digested genomic DNAs showed that the strains belonged to two different clonal lineages, indicating that horizontal transfer of this gene had occurred and suggesting the possibility of further spread of resistance in the future. PMID- 16251288 TI - Identification of a dithiazoline inhibitor of Escherichia coli L,D carboxypeptidase A. AB - The enzyme L,D-carboxypeptidase A is involved in the recycling of bacterial peptidoglycan and is essential in Escherichia coli during stationary phase. By high-throughput screening, we have identified a dithiazoline inhibitor of the enzyme with a 50% inhibitory concentration of 3 microM. The inhibitor appeared to cause lysis of E. coli during stationary phase, behavior that is similar to a previously described deletion mutant of L,D-carboxypeptidase A (M. F. Templin, A. Ursinus, and J.-V. Holtje, EMBO J. 18:4108-4117, 1999). As much as a one-log drop in CFU in stationary phase was observed upon treatment of E. coli with the inhibitor, and the amount of intracellular tetrapeptide substrate increased by approximately 33%, consistent with inhibition of the enzyme within bacterial cells. Stationary-phase targets such as L,D-carboxypeptidase A are largely underrepresented as targets of the antibiotic armamentarium but provide potential opportunities to interfere with bacterial growth and persistence. PMID- 16251287 TI - Relationship of pleconaril susceptibility and clinical outcomes in treatment of common colds caused by rhinoviruses. AB - Pleconaril, a specific inhibitor of human picornaviruses, showed therapeutic efficacy against community-acquired colds caused by rhinoviruses in two placebo controlled trials. Virological assessments were conducted during these trails, including virus culture and drug susceptibility testing. Nasal mucus samples collected from the enrolled patients were tested for the presence of picornavirus by reverse transcriptase PCR and culture. In total, 827 baseline nasal mucus samples were positive by virus culture (420 in the placebo group and 407 in the pleconaril group). Pleconaril treatment was associated with a more rapid loss of culturable virus. By study day 3, the number of samples positive by culture fell to 282 for the placebo-treated subjects and 202 for the pleconaril-treated subjects (P < 0.0001); and by day 6, the number of samples in the two groups positive by culture fell to 196 and 165, respectively (P = 0.07). The clinical benefit correlated strongly with the pleconaril susceptibility of the baseline virus isolate. Pleconaril-treated subjects infected with the more highly susceptible viruses (50% effective concentration < or = 0.38 microg/ml) experienced a median 1.9- to 3.9-day reduction in symptom duration compared with that for the placebo-treated subjects. By contrast, subjects whose baseline virus isolate susceptibility was >0.38 microg/ml did not benefit from pleconaril treatment. These results indicate that the magnitude of symptomatic improvement in pleconaril-treated subjects with community-acquired colds is related to the drug susceptibility of the infecting virus, clearly linking the antiviral effects of the drug to clinical efficacy. Post-baseline virus isolates with reduced susceptibility or full resistance to pleconaril were recovered from 10.7% and 2.7% of drug-treated subjects, respectively. These patients shed low levels of virus and had no unusual clinical outcomes. Nevertheless, studies on the biologic properties and transmissibility of these variant viruses are warranted. PMID- 16251289 TI - Randomized, placebo-controlled trial of nonpegylated and pegylated forms of recombinant human alpha interferon 2a for suppression of dengue virus viremia in rhesus monkeys. AB - Dengue fever and dengue hemorrhagic fever are caused by infection with any one of the four dengue viruses (DVs) and are significant public health burdens throughout the tropics. Higher viremia levels are associated with greater dengue disease severity. A therapeutic intervention to suppress viremia early in DV infection could potentially ameliorate severe disease. Recombinant alpha interferon 2a (rIFN-alpha-2a, Roferon-A) suppressed DV replication in human peripheral blood mononuclear cells in vitro. We therefore examined the effects of rIFN-alpha-2a and pegylated recombinant IFN-alpha-2a (PEG-rIFN-alpha-2a, PEGASYS) on DV serotype 2 (DV-2) viremia in rhesus monkeys. Flavivirus-naive monkeys were inoculated with DV-2 and randomized to receive a single dose of rIFN-alpha-2a (10 million international units/m2) versus placebo or PEG-rIFN-alpha-2a (6 microg/kg) versus placebo 1 day after the onset of viremia. Serial daily viremia levels were measured, and convalescent-phase DV-2 neutralizing antibody titers were determined. Compared to placebo, a single injection of rIFN-alpha-2a temporarily suppressed DV-2 replication and delayed the time to peak viremia by a median of 3 days. However, measures of total viral burden were not different between the two groups. A single injection of PEG-rIFN-alpha-2a significantly lowered daily viremia levels and improved virus clearance, starting 48 h after administration. There were no significant differences in DV-2 neutralizing antibody titers between the treatment and placebo groups at 30 and 90 days postinfection. Based on their individual effects, future studies should investigate a combination of rIFN-alpha-2a and PEG-rIFN-alpha-2a for suppression of dengue virus viremia and as a potential therapeutic intervention. PMID- 16251290 TI - Susceptibilities of antiviral-resistant influenza viruses to novel neuraminidase inhibitors. AB - The susceptibilities of five zanamivir-resistant and six oseltamivir-resistant influenza viruses were assessed against four neuraminidase (NA) inhibitors, including peramivir and A-315675, by a fluorometric NA activity inhibition assay. The enzyme activity of a majority of the variants was effectively inhibited by either A-315675 or both peramivir and A-315675 (50% inhibitory concentration, <10 nM). A novel oseltamivir-resistant influenza virus B variant carrying substitution at residue 198 (Asp-->Asn) (N2 numbering) retained susceptibility to peramivir and A-315675. In vivo, the Asn198 variant showed no apparent fitness impairment as judged by its recovery on day 5 from the nasal washes of ferrets coinfected with equal doses of the wild-type virus and the Asn198 variant. Based on the sequence analysis of the virus in the nasal washes, oseltamivir treatment (5 mg/kg twice daily for 5 days) did not provide growth advantage to the Asn198 variant. Nevertheless, treatment with A-315675 (prodrug A-322278) reduced the number of the animals (two of seven) shedding the Asn198 variant. These studies indicate that different patterns of susceptibility and cross-resistance between NA inhibitors may prove important if antiviral resistance to zanamivir and oseltamivir were to emerge. PMID- 16251291 TI - Use of ribotyping to retrospectively identify methicillin-resistant Staphylococcus aureus isolates from phase 3 clinical trials for tigecycline that are genotypically related to community-associated isolates. AB - A retrospective study was performed to identify methicillin-resistant Staphylococcus aureus (MRSA) isolates obtained from patients enrolled in phase 3 clinical trials for tigecycline that were genotypically similar to known community-associated MRSA (CA-MRSA) strains. The clinical trials were double blind comparator studies for complicated skin and skin structure infections or complicated intra-abdominal infections. We obtained 85% of the MRSA isolates from patients with complicated skin and skin structure infections. Using ribotyping, MRSA isolates were compared with well-characterized North American CA-MRSA strains and negative-control hospital-associated (HA) MRSA strains by cluster analysis; 91 of the 173 isolates clustered with two groups of known CA-MRSA strains, 60% of which shared an indistinguishable ribotype. These isolates were subsequently tested for the presence of SCCmec type IV and the Panton-Valentine leukocidin (PVL)-encoding genes as well as susceptibility to clindamycin, characteristics that are typically associated with CA-MRSA; 89 of the 91 isolates carried the type IV SCCmec element and 76 were also positive for the PVL-encoding genes; 73 of these isolates were susceptible to clindamycin. A similar analysis performed on 26 nonclustering isolates identified only four with these characteristics; 89 of the 91 clustering isolates were inhibited by tigecycline at MICs of < or = 0.5 microg/ml. On the basis of clustering information and preliminary genetic characterization, it appears that ribotyping is a useful tool in identifying potential CA-MRSA isolates and 76 MRSA isolates from patients enrolled in the tigecycline phase 3 trials have genetic markers typically associated with CA-MRSA. PMID- 16251292 TI - Evaluation of antibiotic treatment against "Candidatus Helicobacter suis" in a mouse model. AB - "Helicobacter heilmannii" (proposed name) type 1 colonizes the human stomach. It has been shown to be identical to "Candidatus Helicobacter suis," a Helicobacter species colonizing the stomachs of >60% of slaughter pigs. This bacterium has not been isolated in vitro until now. Antibiotic susceptibility testing of "Candidatus Helicobacter suis" has not been carried out so far. For the present study, a mouse model was adopted to evaluate the antibiotic susceptibility of this organism. Mice infected with "Candidatus Helicobacter suis" were treated with amoxicillin and omeprazole, a therapy which is used to treat H. heilmannii infections in humans. Two different isolates of "Candidatus Helicobacter suis" were tested. The excretion of bacterial DNA was assessed during treatment, using PCR on fecal samples. At the end of the experiment, 8 days after the cessation of treatment, the presence of infection was evaluated using a urease test and a PCR test on stomach samples. A marked decrease in the excretion of bacterial DNA was observed a few days after the onset of treatment, and the level remained low until the end of the experiment. A difference in susceptibility between the two "Candidatus Helicobacter suis" isolates was pointed out. The in vivo mouse model infected with "Candidatus Helicobacter suis" will be useful for further screening of potential therapeutic regimens. PMID- 16251293 TI - Pharmacokinetics, safety, and tolerability of caspofungin in children and adolescents. AB - Caspofungin is a parenteral antifungal that inhibits beta-1,3-D-glucan synthesis. Although licensed for adult use, the appropriate caspofungin dosing regimen in pediatric patients is not yet known. We therefore investigated the pharmacokinetics and safety of caspofungin in pediatric patients. Thirty-nine children (ages 2 to 11 years) and adolescents (ages 12 to 17 years) with neutropenia were administered caspofungin using either a weight-based regimen (1 mg/kg of body weight/day) or a body surface area regimen (50 mg/m2/day or 70 mg/m2/day). Plasma samples for caspofungin profiles were collected on days 1 and 4. These results were compared to those from adults treated with either 50 or 70 mg/day for mucosal candidiasis. In children receiving 1 mg/kg/day (maximum, 50 mg/day), the area under the concentration-time curve over 24 h (AUC(0-24)) was significantly smaller (46% after multiple doses) than that observed in adults receiving 50 mg/day (P < 0.001). In children and adolescents receiving 50 mg/m2/day (maximum, 70 mg/day), the AUC(0-24) following multiple doses was similar to that for the exposure in adults receiving 50 mg/day. The AUC(0-24) and concentration trough (at 24 h) in pediatric patients receiving the 50-mg/m2 daily regimen were consistent across the range of ages. Caspofungin was generally well tolerated in this study. None of the patients developed a serious drug-related adverse event or were discontinued for toxicity. These results demonstrate that caspofungin at 1 mg/kg/day in pediatric patients is suboptimal. Caspofungin administration at 50 mg/m2/day provides a comparable exposure to that of adult patients treated with 50 mg/day. PMID- 16251294 TI - High potency of indolyl aryl sulfone nonnucleoside inhibitors towards drug resistant human immunodeficiency virus type 1 reverse transcriptase mutants is due to selective targeting of different mechanistic forms of the enzyme. AB - Indolyl aryl sulfone (IAS) nonnucleoside inhibitors have been shown to potently inhibit the growth of wild-type and drug-resistant human immunodeficiency virus type 1 (HIV-1), but their exact mechanism of action has not been elucidated yet. Here, we describe the mechanism of inhibition of HIV-1 reverse transcriptase (RT) by selected IAS derivatives. Our results showed that, depending on the substitutions introduced in the IAS common pharmacophore, these compounds can be made selective for different enzyme-substrate complexes. Moreover, we showed that the molecular basis for this selectivity was a different association rate of the drug to a particular enzymatic form along the reaction pathway. By comparing the activities of the different compounds against wild-type RT and the nonnucleoside reverse transcriptase inhibitor-resistant mutant Lys103Asn, it was possible to hypothesize, on the basis of their mechanism of action, a rationale for the design of drugs which could overcome the steric barrier imposed by the Lys103Asn mutation. PMID- 16251295 TI - Prior antimicrobial therapy and risk for hospital-acquired Candida glabrata and Candida krusei fungemia: a case-case-control study. AB - The incidence of infections caused by Candida glabrata and Candida krusei, which are generally more resistant to fluconazole than Candida albicans, is increasing in hospitalized patients. However, the extent to which prior exposure to specific antimicrobial agents increases the risk of subsequent C. glabrata or C. krusei candidemia has not been closely studied. A retrospective case-case-control study was performed at a university hospital. From 1998 to 2003, 60 patients were identified with hospital-acquired non-C. albicans candidemia (C. glabrata or C. krusei; case group 1). For comparison, 68 patients with C. albicans candidemia (case group 2) and a common control group of 121 patients without candidemia were studied. Models were adjusted for demographic and clinical risk factors, and the risk for candidemia associated with exposure to specific antimicrobial agents was assessed. After adjusting for both nonantimicrobial risk factors and receipt of other antimicrobial agents, piperacillin-tazobactam (odds ratio [OR], 4.15; 95% confidence interval [CI], 1.04 to 16.50) and vancomycin (OR, 6.48; CI, 2.20 to 19.13) were significant risk factors for C. glabrata or C. krusei candidemia. For C. albicans candidemia, no specific antibiotics remained a significant risk after adjusted analysis. Prior fluconazole use was not significantly associated with either C. albicans or non-C. albicans (C. glabrata or C. krusei) candidemia. In this single-center study, exposure to antibacterial agents, specifically vancomycin or piperacillin-tazobactam, but not fluconazole, was associated with subsequent hospital-acquired C. glabrata or C. krusei candidemia. Further studies are needed to prospectively analyze specific antimicrobial risks for nosocomial candidemia across multiple hospital centers. PMID- 16251296 TI - Antibacterial activity of human neutrophil defensin HNP-1 analogs without cysteines. AB - The antibacterial activity of human neutrophil defensin HNP-1 analogs without cysteines has been investigated. A peptide corresponding to the HNP-1 sequence without the six cysteines (HNP-1deltaC) exhibited antibacterial activity toward gram-negative and gram-positive bacteria. Truncated analogs wherein the nine N terminal residues of HNP-1 and the remaining three cysteines were deleted (HNP 1deltaC18) or the G was replaced with A (HNP-1deltaC18A) also exhibited antibacterial activity. Substantial activity was observed for HNP-1deltaC and HNP 1deltaC18 in the presence of 100 mM NaCl, except in the case of Pseudomonas aeruginosa. The linear peptides were active in the presence of carbonyl cyanide m chlorophenylhydrazone (CCCP), indicating that proton motive force was not essential for killing of bacteria by the peptides. In fact, in the presence of CCCP, the peptides were active against P. aeruginosa even in the presence of 100 mM NaCl. The antibacterial activity of HNP-1deltaC, but not that of the shorter, 18-residue peptides, was attenuated in the presence of serum. The generation of defensins without cysteines would be easier than that of disulfide-linked defensins. Hence, linear defensins could have potential as therapeutic agents. PMID- 16251297 TI - Pseudomonas aeruginosa AmpR is a global transcriptional factor that regulates expression of AmpC and PoxB beta-lactamases, proteases, quorum sensing, and other virulence factors. AB - In members of the family Enterobacteriaceae, ampC, which encodes a beta lactamase, is regulated by an upstream, divergently transcribed gene, ampR. However, in Pseudomonas aeruginosa, the regulation of ampC is not understood. In this study, we compared the characteristics of a P. aeruginosa ampR mutant, PAOampR, with that of an isogenic ampR+ parent. The ampR mutation greatly altered AmpC production. In the absence of antibiotic, PAOampR expressed increased basal beta-lactamase levels. However, this increase was not followed by a concomitant increase in the P(ampC) promoter activity. The discrepancy in protein and transcription analyses led us to discover the presence of another chromosomal AmpR-regulated beta-lactamase, PoxB. We found that the expression of P. aeruginosa ampR greatly altered the beta-lactamase production from ampC and poxB in Escherichia coli: it up-regulated AmpC but down-regulated PoxB activities. In addition, the constitutive P(ampR) promoter activity in PAOampR indicated that AmpR did not autoregulate in the absence or presence of inducers. We further demonstrated that AmpR is a global regulator because the strain carrying the ampR mutation produced higher levels of pyocyanin and LasA protease and lower levels of LasB elastase than the wild-type strain. The increase in LasA levels was positively correlated with the P(lasA), P(lasI), and P(lasR) expression. The reduction in the LasB activity was positively correlated with the P(rhlR) expression. Thus, AmpR plays a dual role, positively regulating the ampC, lasB, and rhlR expression levels and negatively regulating the poxB, lasA, lasI, and lasR expression levels. PMID- 16251298 TI - Nystatin biosynthesis and transport: nysH and nysG genes encoding a putative ABC transporter system in Streptomyces noursei ATCC 11455 are required for efficient conversion of 10-deoxynystatin to nystatin. AB - The genes nysH and nysG, encoding putative ABC-type transporter proteins, are located at the flank of the nystatin biosynthetic gene cluster in Streptomyces noursei ATCC 11455. To assess the possible roles of these genes in nystatin biosynthesis, they were inactivated by gene replacements leading to in-frame deletions. Metabolite profile analysis of the nysH and nysG deletion mutants revealed that both of them synthesized nystatin at a reduced level and produced considerable amounts of a putative nystatin analogue. Liquid chromatography-mass spectrometry and nuclear magnetic resonance structural analyses of the latter metabolite confirmed its identity as 10-deoxynystatin, a nystatin precursor lacking a hydroxyl group at C-10. Washing experiments demonstrated that both nystatin and 10-deoxynystatin are transported out of cells, suggesting the existence of an alternative efflux system(s) for the transport of nystatin related metabolites. This notion was further corroborated in experiments with the ATPase inhibitor sodium o-vanadate, which affected the production of nystatin and 10-deoxynystatin in the wild-type strain and transporter mutants in a different manner. The data obtained in this study suggest that the efflux of nystatin related polyene macrolides occurs through several transporters and that the NysH NysG efflux system provides conditions favorable for C-10 hydroxylation. PMID- 16251299 TI - TAK-652 inhibits CCR5-mediated human immunodeficiency virus type 1 infection in vitro and has favorable pharmacokinetics in humans. AB - The first small-molecule CCR5 antagonist, TAK-779, could not be developed as an anti-human immunodeficiency virus type (anti-HIV-1) agent because of its poor oral bioavailability. TAK-652 is an orally bioavailable TAK-779 derivative with potent anti-HIV-1 activity. TAK-652 inhibited the binding of RANTES (regulated on activation, normal T-cell expressed and secreted), macrophage inflammatory protein 1alpha (MIP-1alpha), and MIP-1beta to CCR5-expressing cells at nanomolar concentrations. TAK-652 could also suppress the binding of monocyte chemotactic protein 1 (MCP-1) to CCR2b-expressing cells. However, its inhibitory effect on ligand binding to other chemokine receptors was limited. TAK-652 was active against CCR5-using (R5) HIV-1 but totally inactive against CXCR4-using (X4) HIV 1. The compound was active against R5 HIV-1 clinical isolates containing reverse transcriptase and protease inhibitor-resistant mutations, with a mean 50% effective concentration (EC50) and EC90 of 0.061 and 0.25 nM, respectively. In addition, recombinant R5 viruses carrying different subtype (A to G) envelope proteins were equally susceptible to TAK-652. A single oral administration of TAK 652 up to 100 mg was safe and well tolerated in humans. The compound displayed favorable pharmacokinetics, and its plasma concentration was 7.2 ng/ml (9.1 nM) even 24 h after the administration of 25 mg. Thus, TAK-652 is a promising candidate as a novel entry inhibitor of HIV-1. PMID- 16251300 TI - In vitro assessment of methylene blue on chloroquine-sensitive and -resistant Plasmodium falciparum strains reveals synergistic action with artemisinins. AB - Methylene blue (MB) represents a promising antimalarial drug candidate for combination therapies against drug-resistant parasite strains. To support and facilitate the application of MB in future field trials, we studied its antiparasitic effects in vitro. MB is active against all blood stages of both chloroquine (CQ)-sensitive and CQ-resistant P. falciparum strains with 50% inhibitory concentration (IC50) values in the lower nanomolar range. Ring stages showed the highest susceptibility. As demonstrated by high-performance liquid chromatography-tandem mass spectrometry on different cell culture compartments, MB is accumulated in malarial parasites. In drug combination assays, MB was found to be antagonistic with CQ and other quinoline antimalarials like piperaquine and amodiaquine; with mefloquine and quinine, MB showed additive effects. In contrast, we observed synergistic effects of MB with artemisinin, artesunate, and artemether for all tested parasite strains. Artemisinin/MB combination concentration ratios of 3:1 were found to be advantageous, demonstrating that the combination of artemisinin with a smaller amount of MB can be recommended for reaching maximal therapeutic effects. Our in vitro data indicate that combinations of MB with artemisinin and related endoperoxides might be a promising option for treating drug-resistant malaria and should be studied in future field trials. Resistance development under this drug combination is unlikely to occur. PMID- 16251301 TI - Phosphinothricin tripeptide synthetases in Streptomyces viridochromogenes Tu494. AB - The tripeptide backbone of phosphinothricin (PT) tripeptide (PTT), a compound with herbicidal activity from Streptomyces viridochromogenes, is assembled by three stand-alone peptide synthetase modules. The enzyme PhsA (66 kDa) recruits the PT-precursor N-acetyl-demethylphosphinothricin (N-Ac-DMPT), whereas the two alanine residues of PTT are assembled by the enzymes PhsB and PhsC (129 and 119 kDa, respectively). During or after assembly, the N-Ac-DMPT residue in the peptide is converted to PT by methylation and deacetylation. Both phsB and phsC appear to be cotranscribed together with two other genes from a single promoter and they are located at a distance of 20 kb from the gene phsA, encoding PhsA, in the PTT biosynthesis gene cluster of S. viridochromogenes. PhsB and PhsC represent single nonribosomal peptide synthetase elongation modules lacking a thioesterase domain. Gene inactivations, genetic complementations, determinations of substrate specificity of the heterologously produced proteins, and comparison of PhsC sequence with the amino terminus of the alanine-activating nonribosomal peptide synthetase PTTSII from S. viridochromogenes confirmed the role of the two genes in the bialanylation of Ac-DMPT. The lack of an integral thioesterase domain in the PTT assembly system points to product release possibly involving two type II thioesterase genes (the1 and the2) located in the PTT gene cluster alone or in conjunction with an as yet unknown mechanism of product release. PMID- 16251302 TI - Mechanisms of azole resistance in a clinical isolate of Candida tropicalis. AB - Azole resistance has been insufficiently investigated in the yeast Candida tropicalis. Here we determined the molecular mechanisms responsible for azole resistance in a clinical isolate of this pathogenic yeast. Antifungal susceptibility testing performed by a disk diffusion method showed resistance or markedly decreased susceptibility to azoles, which was confirmed by determination of MICs. Considering the relationship between azole susceptibility and the respiration reported for other yeast species, the respiratory activity of this isolate was investigated. Flow cytometry using rhodamine 123 and oxygraphy demonstrated an increased respiratory activity, which was not linked to an overexpression or increased number of copies of the mitochondrial genome. Among previously described resistance mechanisms, an increased activity of efflux pumps was investigated by flow cytometry using rhodamine 6G. However, the efflux of rhodamine 6G was lower in the resistant isolate than in susceptible ones. Likewise, real-time reverse transcription-PCR quantification of the expression of C. tropicalis MDR1 (CtMDR1), which encodes an efflux protein belonging to the major facilitator superfamily, did not show overexpression of this gene. In contrast, the resistant isolate overexpressed the CtERG11 gene coding for lanosterol 14alpha-demethylase. This was in agreement with the larger amount of ergosterol found in this isolate. Moreover, sequencing of CtERG11 showed a point mutation leading to a tyrosine substitution in the protein sequence, which might lead to decreased binding affinity for azoles. In conclusion, overexpression of CtERG11 associated with a missense mutation in this gene seemed to be responsible for the acquired azole resistance of this clinical isolate. PMID- 16251303 TI - Gene expression analysis of the mechanism of natural Sb(V) resistance in Leishmania donovani isolates from Nepal. AB - Control of visceral leishmaniasis (VL) is being challenged by the emergence of natural resistance against the first line of treatment, pentavalent antimonials [Sb(V)]. An insight into the mechanism of natural Sb(V) resistance is required for the development of efficient strategies to monitor the emergence and spreading of Sb(V) resistance in countries where VL is endemic. In this work, we have focused on the mechanism of natural Sb(V) resistance emerging in Nepal, a site where anthroponotic VL is endemic. Based on the current knowledge of Sb(V) metabolism and of the in vitro trivalent antimonial [Sb(III)] models of resistance to Leishmania spp., we selected nine genes for a comparative transcriptomic study on natural Sb(V)-resistant and -sensitive Leishmania donovani isolates. Differential gene expression patterns were observed for the genes coding for 2-thiol biosynthetic enzymes, gamma-glutamylcysteine synthetase (GCS) and ornithine decarboxylase (ODC), and for the Sb(III) transport protein aquaglyceroporin 1 (AQP1). The results indicate that the mechanism for natural Sb(V) resistance partially differs from the mechanism reported for in vitro Sb(III) resistance. More specifically, we hypothesize that natural Sb(V) resistance results from (i) a changed thiol metabolism, possibly resulting in inhibition of Sb(V) activation in amastigotes, and (ii) decreased uptake of the active drug Sb(III) by amastigotes. PMID- 16251304 TI - Amino acid mutations essential to production of an altered PBP 2X conferring high level beta-lactam resistance in a clinical isolate of Streptococcus pneumoniae. AB - Altered penicillin-binding protein 2X (PBP 2X) is a primary beta-lactam antibiotic resistance determinant and is essential to the development of penicillin and cephalosporin resistance in the pneumococcus. We have studied the importance for resistance of 23 amino acid substitutions located in the transpeptidase domain (TD) of PBP 2X from an isolate with high-level resistance, isolate 3191 (penicillin MIC, 16 mug/ml; cefotaxime MIC, 4 microg/ml). Strain R6(2X/2B/1A/mur) (for which the MICs are as described for isolate 3191) was constructed by transforming laboratory strain R6 with all the necessary resistance determinants (altered PBPs 2X, 2B, and 1A and altered MurM) from isolate 3191. Site-directed mutagenesis was used to reverse amino acid substitutions in altered PBP 2X, followed by investigation of the impact of these reversions on resistance levels in R6(2X/2B/1A/mur). Of the 23 substitutions located in the TD of PBP 2X, reversals at six positions decreased the resistance levels in R6(2X/2B/1A/mur). Reversal of the Thr338Pro and Ile371Thr substitutions individually decreased the penicillin and cefotaxime MICs to 2 and 1 microg/ml, respectively, and individually displayed the greatest impact on resistance. To a lesser extent, reversal of the Leu364Phe, Ala369Val, Arg384Gly, and Tyr595Phe substitutions individually also decreased the penicillin and cefotaxime MICs. Reversal at all six positions collectively decreased both the penicillin and the cefotaxime MICs of R6(2X/2B/1A/mur) to 0.06 microg/ml. This study confirms the essential role of altered PBP 2X as a resistance determinant. Our data reveal that, for isolate 3191, the six amino acid substitutions described above are collectively essential to the production of an altered PBP 2X required for high level resistance to penicillin and cefotaxime. PMID- 16251305 TI - Inhibitory activities of epidermal growth factor receptor tyrosine kinase targeted dihydroxyisoflavone and trihydroxydeoxybenzoin derivatives on Sarcocystis neurona, Neospora caninum, and Cryptosporidium parvum development. AB - Several gene sequences of parasitic protozoa belonging to protein kinase gene families and epidermal growth factor (EGF)-like peptides, which act via binding to receptor tyrosine kinases of the EGF receptor (EGFR) family, appear to mediate host-protozoan interactions. As a clue to EGFR protein tyrosine kinase (PTK) mediation and a novel approach for identifying anticoccidial agents, activities against Sarcocystis neurona, Neospora caninum, and Cryptosporidium parvum grown in BM and HCT-8 cell cultures of 52 EGFR PTK inhibitor isoflavone analogs (dihydroxyisoflavone and trihydroxydeoxybenzoine derivatives) were investigated. Their cytotoxicities against host cells were either absent, mild, or moderate by a nitroblue tetrazolium test. At concentrations ranging from 5 to 10 microg/ml, 20 and 5 analogs, including RM-6427 and RM-6428, exhibited an in vitro inhibitory effect of > or = 95% against at least one parasite or against all three, respectively. In immunosuppressed Cryptosporidium parvum-infected Mongolian gerbils orally treated with either 200 or 400 mg of agent RM-6427/kg of body weight/day for 8 days, fecal microscopic oocyst shedding was abolished in 6/10 animals (P of <0.001 versus untreated controls) and mean shedding was reduced by 90.5% (P of <0.0001) and 92.0% (P of <0.0001), respectively, higher levels of inhibition than after nitazoxanide (200 mg/kg/day for 8 days) or paromomycin (100 mg/kg/day for 8 days) treatment (55.0%, P of <0.001, and 17.5%, P of >0.05, respectively). After RM-6427 therapy (200 mg/kg/day for 8 days), the reduction in the ratio of animals with intracellular parasites was nearly significant in ileum (P = 0.067) and more marked in the biliary tract (P < 0.0013) than after nitazoxanide or paromomycin treatment (0.05 < P < 0.004). RM-6428 treatment at a regimen of 400 mg/kg/day for 12 days inhibited oocyst shedding, measured using flow cytometry from day 4 (P < 0.05) to day 12 (P < 0.02) of therapy, when 2/15 animals had no shedding (P < 0.0001) and 11/15 were free of gut and/or biliary tract parasites (P < 0.01). No mucosal alteration was microscopically observed for treated or untreated infected gerbils. To our knowledge, this report is the first to suggest that the isoflavone class of agents has the potential for anticoccidial therapy. PMID- 16251306 TI - Expression of the mef(E) gene encoding the macrolide efflux pump protein increases in Streptococcus pneumoniae with increasing resistance to macrolides. AB - Active macrolide efflux is a major mechanism of macrolide resistance in Streptococcus pneumoniae in many parts of the world, especially North America. In Canada, this active macrolide efflux in S. pneumoniae is predominantly due to acquisition of the mef(E) gene. In the present study, we assessed the mef(E) gene sequence as well as mef(E) expression in variety of low- and high-level macrolide resistant, clindamycin-susceptible (M-phenotype) S. pneumoniae isolates (erythromycin MICs, 1 to 32 microg/ml; clindamycin MICs, < or = 0.25 microg/ml). Southern blot hybridization with mef(E) probe and EcoRI digestion and relative real-time reverse transcription-PCR were performed to study the mef(E) gene copy number and expression. Induction of mef(E) expression was analyzed by Etest susceptibility testing pre- and postincubation with subinhibitory concentrations of erythromycin, clarithromycin, azithromycin, telithromycin, and clindamycin. The macrolide efflux gene, mef(E), was shown to be a single-copy gene in all 23 clinical S. pneumoniae isolates tested, and expression post-macrolide induction increased 4-, 6-, 20-, and 200-fold in isolates with increasing macrolide resistance (erythromycin MICs 2, 4, 8, and 32 microg/ml, respectively). Sequencing analysis of the macrolide efflux genetic assembly (mega) revealed that mef(E) had a 16-bp deletion 153 bp upstream of the putative start codon in all 23 isolates. A 119-bp intergenic region between mef(E) and mel was sequenced, and a 99-bp deletion was found in 11 of the 23 M-phenotype S. pneumoniae isolates compared to the published mega sequence. However, the mef(E) gene was fully conserved among both high- and low-level macrolide-resistant isolates. In conclusion, increased expression of mef(E) is associated with higher levels of macrolide resistance in macrolide-resistant S. pneumoniae. PMID- 16251307 TI - Horizontal transfer of iturin A operon, itu, to Bacillus subtilis 168 and conversion into an iturin A producer. AB - Iturin A and its derivatives are lipopeptide antibiotics produced by Bacillus subtilis and several closely related bacteria. Three iturin group operons (i.e., iturin A, mycosubtilin, and bacillomycin D) of those antibiotic-producing strains have been cloned and sequenced thus far, strongly implying the horizontal transfer of these operons. To examine the nature of such horizontal transfer in terms of antibiotic production, a 42-kb region of the B. subtilis RB14 genome, which contains a complete 38-kb iturin A operon, was transferred via competent cell transformation to the genome of a non-iturin A producer, B. subtilis 168, using a method based on double-crossover homologous recombination with two short landing pad sequences (LPSs) in the genome. The recombinant was positively selected by confirming the elimination of the cI repressor gene, which was localized between the two LPSs and substituted by the transferred segment. The iturin A operon-transferred strain 168 was then converted into an iturin A producer by the introduction of an sfp gene, which encodes 4'-phosphopantetheinyl transferase and is mutated in strain 168. By inserting the pleiotropic regulator degQ, the productivity of iturin A increased sevenfold and was restored to about half that of the donor strain RB14, without the transfer of additional genes, such as regulatory or self-resistance genes. PMID- 16251308 TI - A class of pantothenic acid analogs inhibits Plasmodium falciparum pantothenate kinase and represses the proliferation of malaria parasites. AB - The growth and proliferation of the human malaria parasite Plasmodium falciparum are dependent on the parasite's ability to obtain essential nutrients. One nutrient for which the parasite has an absolute requirement is the water-soluble vitamin pantothenic acid (vitamin B5). In this study, a series of pantothenic acid analogs which retain the 2,4-dihydroxy-3,3-dimethylbutyramide core of pantothenic acid but deviate in structure from one another and from pantothenic acid in the nature of the substituent attached to the amide nitrogen were synthesized using an efficient single-step synthetic route. Eight of 10 analogs tested inhibited the proliferation of intraerythrocytic P. falciparum parasites in vitro, doing so with 50% inhibitory concentrations between 15 and 200 microM. The compounds were generally selective, inhibiting the proliferation of a human cell line (the Jurkat cell line) only at concentrations severalfold higher than those required for inhibition of parasite growth. It was demonstrated that compounds in this series inhibited the phosphorylation of pantothenic acid by pantothenate kinase, the first step in the parasite's biosynthesis of the essential enzyme cofactor coenzyme A, doing so competitively, with K(i) values in the nanomolar range. PMID- 16251309 TI - Safety and efficacy of tigecycline in treatment of skin and skin structure infections: results of a double-blind phase 3 comparison study with vancomycin aztreonam. AB - In a randomized, double-blind, controlled trial, 546 patients with complicated skin and skin structure infections received tigecycline 100 mg/day (a 100-mg initial dose and then 50 mg intravenously twice daily) or the combination of vancomycin 2 g/day (1 g intravenously twice daily) and aztreonam 4 g/day (2 g intravenously twice daily) for up to 14 days. The primary end point was the clinical response in the clinical modified intent-to-treat (c-mITT) and clinically evaluable (CE) populations at the test-of-cure visit 12 to 92 days after the last dose. The microbiologic response at the test-of-cure visit was also assessed. Safety was assessed by physical examination, laboratory results, and adverse event reporting. Five hundred twenty patients were included in the c mITT population (tigecycline group, n = 261; combination group, n = 259), and 436 were clinically evaluable (tigecycline group, n = 223; combination group, n = 213). The clinical responses in the tigecycline and the combination vancomycin and aztreonam groups were similar in the c-mITT population (84.3% versus 86.9%; difference, -2.6% [95% confidence interval, -9.0, 3.8]; P = 0.4755) and the CE population (89.7% versus 94.4%; difference, -4.7% [95% confidence interval, 10.2, 0.8]; P = 0.1015). Microbiologic eradication (documented or presumed) occurred in 84.8% of the patients receiving tigecycline and 93.2% of the patients receiving vancomycin and aztreonam (difference, -8.5 [95% confidence interval, 16.0, -1.0]; P = 0.0243). The numbers of patients reporting adverse events were similar in the two groups, with increased nausea and vomiting rates in the tigecycline group and an increased incidence of rash and increases in alanine aminotransferase and aspartate aminotransferase levels in the combination vancomycin and aztreonam group. Tigecycline was shown to be safe and effective for the treatment of complicated skin and skin structure infections. PMID- 16251310 TI - Virulence factors of Escherichia coli isolates that produce CTX-M-type extended spectrum beta-lactamases. AB - This study determined the phylogenetic groups and virulence factors of 37 Escherichia coli isolates producing types of CTX-M compared with those of 19 isolates producing different types of extended-spectrum beta-lactamases (ESBLs) in a well-defined North American population. Most CTX-M-14 producers (97%) were from phylogenic group D; 67% of the CTX-M-15 producers were from group B2. A single CTX-M-14-producing strain belonged to clonal group A. There were significant prevalence differences for individual virulence factors among CTX-M producers and nonproducers; however, aggregate virulence factor scores were similar. CTX-M producers more commonly caused repeat urinary tract infections. Our results indicate that CTX-M type predicts phylogenetic background, and the virulence potential of ESBL-producing E. coli isolates is a complex issue, requiring further study and ongoing surveillance. PMID- 16251311 TI - Organotypic epithelial raft cultures as a model for evaluating compounds against alphaherpesviruses. AB - The course of herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) and varicella-zoster virus (VZV) infections in squamous epithelial cells cultured in a three-dimensional organotypic raft culture was tested. In these raft cultures, normal human keratinocytes isolated from neonatal foreskins grown at the air liquid interface stratified and differentiated, reproducing a fully differentiated epithelium. Typical cytopathic changes identical to those found in the squamous epithelium in vivo, including ballooning and reticular degeneration with the formation of multinucleate cells, were observed throughout the raft following infection with HSV and VZV at different times after lifting the cultures to the air-liquid interface. For VZV, the aspects of the lesions depended on the stage of differentiation of the organotypic cultures. The activity of reference antiviral agents, acyclovir (ACV), penciclovir (PCV), brivudin (BVDU), foscarnet (PFA), and cidofovir (CDV), was evaluated against wild type and thymidine kinase (TK) mutants of HSV and VZV in the raft cultures. ACV, PCV, and BVDU protected the epithelium against cytopathic effect induced by wild type viruses in a concentration-dependent manner, while treatment with CDV and PFA proved protective against the cytodestructive effects induced by both TK+ and TK- strains. The quantification of the antiviral effects in the rafts were accomplished by measuring viral titers by plaque assay for HSV and by measuring viral DNA load by real-time PCR for VZV. A correlation between the degree of protection as determined by histological examination and viral quantification could be demonstrated The three-dimensional epithelial raft culture represents a novel model for the study of antiviral agents active against HSV and VZV. Since no animal model is available for the evaluation of antiviral agents against VZV, the organotypic cultures may be considered a model to evaluate the efficacy of new anti-VZV antivirals before clinical trials. PMID- 16251312 TI - DNA sequence and comparative genomics of pAPEC-O2-R, an avian pathogenic Escherichia coli transmissible R plasmid. AB - In this study, a 101-kb IncF plasmid from an avian pathogenic Escherichia coli (APEC) strain (APEC O2) was sequenced and analyzed, providing the first completed APEC plasmid sequence. This plasmid, pAPEC-O2-R, has functional transfer and antimicrobial resistance-encoding regions. The resistance-encoding region encodes resistance to eight groups of antimicrobial agents, including silver and other heavy metals, quaternary ammonium compounds, tetracycline, sulfonamides, aminoglycosides, trimethoprim, and beta-lactam antimicrobial agents. This region of the plasmid is unique among previously described IncF plasmids in that it possesses a class 1 integron that harbors three gene cassettes and a heavy metal resistance operon. This region spans 33 kb and is flanked by the RepFII plasmid replicon and an assortment of plasmid maintenance genes. pAPEC-O2-R also contains a 32-kb transfer region that is nearly identical to that found in the E. coli F plasmid, rendering it transferable by conjugation to plasmid-less strains of bacteria, including an APEC strain, a fecal E. coli strain from an apparently healthy bird, a Salmonella enterica serovar Typhimurium strain, and a uropathogenic E. coli strain from humans. Differences in the G+C contents of individual open reading frames suggest that various regions of pAPEC-O2-R had dissimilar origins. The presence of pAPEC-O2-R-like plasmids that encode resistance to multiple antimicrobial agents and that are readily transmissible from APEC to other bacteria suggests the possibility that such plasmids may serve as a reservoir of resistance genes for other bacteria of animal and human health significance. PMID- 16251313 TI - Mouse adenovirus type 1 infection in SCID mice: an experimental model for antiviral therapy of systemic adenovirus infections. AB - The importance of human adenovirus infections in immunocompromised patients urges for new and adequate antiadenovirus compounds. Since human adenoviruses are species specific, animal models for systemic adenovirus infections rely on a nonhuman adenovirus. We established mouse adenovirus type 1 (MAV-1) infection of BALB/c SCID mice as a model for the evaluation of antiadenovirus therapy. In vitro studies with mouse embryonic fibroblasts pointed to the acyclic nucleoside phosphonate cidofovir and the N-7-substituted acyclic derivative 2-amino-7-(1,3 dihydroxy-2-propoxymethyl)purine (S-2242) as markedly active compounds against MAV-1. SCID mice, infected intranasally with MAV-1, developed a fatal disseminated infection after approximately 19 days, characterized by hemorrhagic enteritis. Several techniques were optimized to monitor viral, immunological, and pathological aspects of MAV-1 infection. Real-time PCR quantification of viral DNA revealed that after replication in the lungs, virus disseminated to several organs, including the brain, liver, spleen, intestine, heart, and kidneys (resulting in viruria). Immunohistochemical staining showed that MAV-1 was localized in the endothelial cells of the affected organs. Using reverse transcription-PCR, tissue levels of proinflammatory cytokines (i.e., interleukin 1beta and tumor necrosis factor alpha) were found to be markedly increased. The MAV-1/SCID model appears to be an appropriate model for in vivo evaluation of antiadenovirus agents. Treatment with cidofovir or S-2242 at a dose of 100 mg per kg of body weight resulted in a significant delay in MAV-1-related death, although these antivirals were unable to completely suppress virus replication despite continued drug treatment. These findings suggest that complete virus clearance during antiviral therapy for disseminated adenovirus infection may require an efficient adaptive immune response from the host. PMID- 16251314 TI - Comparative effects of two neutralizing anti-respiratory syncytial virus (RSV) monoclonal antibodies in the RSV murine model: time versus potency. AB - Respiratory syncytial virus (RSV) is the leading viral pathogen responsible for bronchiolitis and pneumonia in infants and young children worldwide. We have previously shown in the mouse model that treatment with an anti-RSV neutralizing monoclonal antibody (MAb) against the F glycoprotein of RSV, palivizumab, decreased lung inflammation, airway obstruction, and postmethacholine airway hyperresponsiveness. MEDI-524, or Numax, is a new MAb derived from palivizumab with enhanced neutralizing activity against RSV. We compared the effects of these two MAbs on different markers of disease severity using the murine model of RSV infection. BALB/c mice were intranasally inoculated with RSV A2. Palivizumab or MEDI-524 was administered once at either 24 h before or 48 h after RSV inoculation. Regardless of the time of administration, all treated mice showed significantly decreased RSV loads in bronchoalveolar lavage samples measured by plaque assay. Only MEDI-524 given at -24 h significantly decreased lung RSV RNA loads on days 5 and 28 after RSV inoculation. Pulmonary histopathologic scores, airway obstruction, and postmethacholine airway hyperresponsiveness were significantly reduced in mice treated with MEDI-524 at 24 h before inoculation, compared with untreated controls and the other regimens evaluated. MEDI-524 was superior to palivizumab on several outcome variables of RSV disease assessed in the mouse model: viral replication, inflammatory and clinical markers of acute disease severity, and long-term pulmonary abnormalities. PMID- 16251315 TI - Analysis of binding sites for the new small-molecule CCR5 antagonist TAK-220 on human CCR5. AB - G protein-coupled receptor CCR5 is the main coreceptor for macrophage-tropic human immunodeficiency virus type 1 (HIV-1), and various small-molecule CCR5 antagonists are being developed to treat HIV-1 infection. It has been reported that such CCR5 antagonists, including TAK-779, bind to a putative binding pocket formed by transmembrane domains (TMs) 1, 2, 3 and 7 of CCR5, indicating the importance of the conformational changes of the TMs during virus entry. In this report, using a single-round infection assay with human CCR5 and its substitution mutants, we demonstrated that a new CCR5 antagonist, TAK-220, shares the putative interacting amino acid residues Asn252 and Leu255 in TM6 with TAK-779 but also requires the distinct residues Gly163 and Ile198 in TMs 4 and 5, respectively, for its inhibitory effect. We suggested that, together with molecular models of the interactions between the drugs and CCR5, the inhibitory activity of TAK-220 could involve direct interactions with amino acid residues in TMs 4, 5, and 6 in addition to those in the previously postulated binding pocket. The possible interaction of drugs with additional regions of the CCR5 molecule would help to develop a new small-molecule CCR5 antagonist. PMID- 16251316 TI - Molecular analysis of the simultaneous production of two SHV-type extended spectrum beta-lactamases in a clinical isolate of Enterobacter cloacae by using single-nucleotide polymorphism genotyping. AB - Bacteria that simultaneously produce multiple extended-spectrum beta-lactamases are frequently isolated. We report an Enterobacter cloacae isolate, ES24, producing four different beta-lactamases (AmpC type beta-lactamase, TEM-1, SHV-7, and a novel extended-spectrum beta-lactamase, SHV-30). Direct sequencing of bla(SHV) gene products gave a "double peak" at position 703, suggesting the presence of more than one allele. Using fluorescence resonance energy transfer real-time PCR to detect single-nucleotide polymorphisms, we were able to distinguish two different bla(SHV) genes in a single isolate. This may prove to be a useful technique in surveys of beta-lactamase production in contemporary clinical isolates. PMID- 16251317 TI - Maraviroc (UK-427,857), a potent, orally bioavailable, and selective small molecule inhibitor of chemokine receptor CCR5 with broad-spectrum anti-human immunodeficiency virus type 1 activity. AB - Maraviroc (UK-427,857) is a selective CCR5 antagonist with potent anti-human immunodeficiency virus type 1 (HIV-1) activity and favorable pharmacological properties. Maraviroc is the product of a medicinal chemistry effort initiated following identification of an imidazopyridine CCR5 ligand from a high-throughput screen of the Pfizer compound file. Maraviroc demonstrated potent antiviral activity against all CCR5-tropic HIV-1 viruses tested, including 43 primary isolates from various clades and diverse geographic origin (geometric mean 90% inhibitory concentration of 2.0 nM). Maraviroc was active against 200 clinically derived HIV-1 envelope-recombinant pseudoviruses, 100 of which were derived from viruses resistant to existing drug classes. There was little difference in the sensitivity of the 200 viruses to maraviroc, as illustrated by the biological cutoff in this assay (= geometric mean plus two standard deviations [SD] of 1.7 fold). The mechanism of action of maraviroc was established using cell-based assays, where it blocked binding of viral envelope, gp120, to CCR5 to prevent the membrane fusion events necessary for viral entry. Maraviroc did not affect CCR5 cell surface levels or associated intracellular signaling, confirming it as a functional antagonist of CCR5. Maraviroc has no detectable in vitro cytotoxicity and is highly selective for CCR5, as confirmed against a wide range of receptors and enzymes, including the hERG ion channel (50% inhibitory concentration, >10 microM), indicating potential for an excellent clinical safety profile. Studies in preclinical in vitro and in vivo models predicted maraviroc to have human pharmacokinetics consistent with once- or twice-daily dosing following oral administration. Clinical trials are ongoing to further investigate the potential of using maraviroc for the treatment of HIV-1 infection and AIDS. PMID- 16251318 TI - Molecular mechanisms of beta-lactam resistance mediated by AmpC hyperproduction in Pseudomonas aeruginosa clinical strains. AB - The molecular mechanisms of beta-lactam resistance mediated by AmpC hyperproduction in natural strains of Pseudomonas aeruginosa were investigated in a collection of 10 isogenic, ceftazidime-susceptible and -resistant pairs of isolates, each sequentially recovered from a different intensive care unit patient treated with beta-lactams. All 10 ceftazidime-resistant mutants hyper produced AmpC (beta-lactamase activities were 12- to 657-fold higher than those of the parent strains), but none of them harbored mutations in ampR or the ampC ampR intergenic region. On the other hand, six of them harbored inactivating mutations in ampD: four contained frameshift mutations, one had a C-->T mutation, creating a premature stop codon, and finally, one had a large deletion, including the complete ampDE region. Complementation studies revealed that only three of the six ampD mutants could be fully trans-complemented with either ampD- or ampDE harboring plasmids, whereas one of them could be trans-complemented only with ampDE and two of them (including the mutant with the deletion of the ampDE region and one with an ampD frameshift mutation leading to an ampDE-fused open reading frame) could not be fully trans-complemented with any of the plasmids. Finally, one of the four mutants with no mutations in ampD could be trans-complemented, but only with ampDE. Although the inactivation of AmpD is found to be the most frequent mechanism of AmpC hyperproduction in clinical strains, our findings suggest that for certain types of mutations, AmpE plays an indirect role in resistance and that there are other unknown genes involved in AmpC hyperproduction, with at least one of them apparently located close to the ampDE operon. PMID- 16251319 TI - Spontaneous conversion to quinolone and fluoroquinolone resistance among wild type Escherichia coli isolates in relation to phylogenetic background and virulence genotype. AB - Human clinical isolates of Escherichia coli that are resistant to quinolone or fluoroquinolone agents typically exhibit fewer extraintestinal virulence factors (VFs) than susceptible isolates, along with a different phylogenetic background. To experimentally assess the basis for this as-yet-unexplained phenomenon, 40 E. coli strains (20 E. coli Reference collection members and 20 Israeli cystitis isolates) were subjected to serial selective passaging to obtain derivatives resistant to nalidixic acid (NA) and ciprofloxacin (C). PCR-based VF profiling and phylotyping were performed on the parents and their respective resistant derivatives. All 40 susceptible parent strains yielded NA- and C-resistant derivatives after a median of 6 (range, 4 to 12) serial selective passages on agar plates containing increasing concentrations of NA and C. The numbers of passages required for resistance did not differ by collection origin, phylogenetic group, basal VF profile, source (urine versus fecal), or host group (human versus animal). With the development of C resistance, only one VF was lost in a single strain. Resistant derivatives exhibited the same phylotype as their susceptible parents. These findings suggest that the sparse VF profiles and the low-virulence phylogenetic background of NA- and C-resistant E. coli clinical isolates probably are not attributable to the loss of VFs from intrinsically high virulence strains during conversion to resistance or to enhanced emergence of drug resistance among intrinsically low-virulence strains. A more likely explanation is the importation of resistant strains from an as-yet-undefined low virulence external selection reservoir. PMID- 16251320 TI - CTX-M-2 and a new CTX-M-39 enzyme are the major extended-spectrum beta-lactamases in multiple Escherichia coli clones isolated in Tel Aviv, Israel. AB - The rate of occurrence of the extended-spectrum beta-lactamase (ESBL)-producing phenotype among Escherichia coli isolates in Tel Aviv is 12% (22). The aim of this study was to understand the molecular epidemiology of E. coli ESBL producers and to identify the ESBL genes carried by them. We studied 20 single-patient ESBL producing E. coli clinical isolates. They comprised 11 distinct nonrelated pulsed field gel electrophoresis (PFGE) genotypes: six isolates belonged to the same PFGE clone, four other clones included two isolates each, and six unrelated clones included only one isolate. All isolates produced various beta-lactamases with pIs ranging from 5.2 to 8.2, varying within similar PFGE clones. The most prevalent ESBL gene was bla(CTX-M); 16 isolates carried bla(CTX-M-2) and three carried a new ESBL gene designated bla(CTX-M-39). Three strains carried bla(SHV) (two bla(SHV-12) and one bla(SHV-5)), and two strains carried inhibitor-resistant ESBL genes, bla(TEM-33) and bla(TEM-30); 18 strains carried bla(TEM-1) and eight strains carried bla(OXA-2). Plasmid mapping and Southern blot analysis with a CTX M-2 probe demonstrated that bla(CTX-M-2) is plasmid borne. The wide dissemination of ESBLs among E. coli isolates in our institution is partly related to clonal spread, but more notably to various plasmid-associated ESBL genes, occurring in multiple clones, wherein the CTX-M gene family appears almost uniformly. We report here a new CTX-M gene, designated bla(CTX-M-39), which revealed 99% homology with bla(CTX-M-26), with a substitution of arginine for glutamine at position 225. PMID- 16251321 TI - Combination therapy with terbinafine and amphotericin B in a rabbit model of experimental invasive aspergillosis. AB - Antagonistic effects of combination therapy using amphotericin B (AmB) with agents which block ergosterol synthesis are a concern. Terbinafine was evaluated with AmB to assess antagonism or synergy in a rabbit model of invasive aspergillosis. Terbinafine had relatively little activity but did not demonstrate antagonism against AmB in our model. PMID- 16251322 TI - Increased mutation frequencies in Escherichia coli isolates harboring extended spectrum beta-lactamases. AB - Hypermutable (mutation frequency [f], > or = 4 x 10(-8)) Escherichia coli strains were more frequently found (43%) in a collection of 89 extended-spectrum beta lactamase (ESBL)-producing isolates from different patients (77 pulsed-field gel electrophoresis clones, 12 ESBL types) than in non-ESBL E. coli (26%) strains (P = 0.03). Among urinary tract isolates, the frequency of hypermutation was 40% in ESBL versus 26% in non-ESBL isolates (P = 0.03). PMID- 16251323 TI - Activities of flucytosine, fluconazole, amphotericin B, and micafungin in a murine model of disseminated infection by Candida glabrata. AB - We compared the efficacies of amphotericin B, fluconazole, flucytosine, and micafungin in a systemic murine infection by three isolates of Candida glabrata. Amphotericin B showed the best results, although none of the drugs dramatically reduced mortality or tissue burden in liver or spleen. PMID- 16251324 TI - Kinetics study of KPC-3, a plasmid-encoded class A carbapenem-hydrolyzing beta lactamase. AB - The kinetic activity of KPC-3, a plasmid-encoded class A carbapenemase, was studied. It hydrolyzed penicillins, cephalosporins, carbapenems, and even sulbactam. The best substrate was cephalothin (k(cat/K)m = 3.48 microM(-1) s( 1)). The efficiency of the enzyme was similar for imipenem and meropenem (k(cat)/K(m), 1.4 and 1.94 microM(-1) s(-1), respectively). PMID- 16251325 TI - AmpC and OprD are not involved in the mechanism of imipenem hypersusceptibility among Pseudomonas aeruginosa isolates overexpressing the mexCD-oprJ efflux pump. AB - Pseudomonas aeruginosa strains that overexpress mexCD-oprJ become hypersusceptible to imipenem. Disruption of AmpC induction has been suggested to cause this phenotype. However, data from this study demonstrate that hypersusceptibility to imipenem can develop without changes in ampC expression or AmpC activity. Furthermore, hypersusceptibility is not caused by changes in expression of the outer membrane porin, OprD. PMID- 16251326 TI - TLA-2, a novel Ambler class A expanded-spectrum beta-lactamase. AB - Beta-lactamase TLA-2 is encoded by a 47-kb plasmid isolated from an unidentified bacterial strain from a wastewater treatment plant. TLA-2 is an Ambler class A beta-lactamase that shares 52% amino acid identity with CGA-1 from Chryseobacterium gleum and 51% with TLA-1 from Escherichia coli. The enzyme hydrolyzes mostly cephalosporins. PMID- 16251327 TI - Recombination between the dfrA12-orfF-aadA2 cassette array and an aadA1 gene cassette creates a hybrid cassette, aadA8b. AB - Homologous recombination between closely related gene cassettes, such as aadA1 and aadA2, which are 89% identical, can create hybrid cassettes and hybrids of existing cassette arrays. A new cassette array, dfrA12-orfF-aadA8b, which was created by such a recombination event occurring within the aadA2 cassette in the dfrA12-orfF-aadA2 array, has been identified. PMID- 16251328 TI - mmpL7 gene of Mycobacterium tuberculosis is responsible for isoniazid efflux in Mycobacterium smegmatis. AB - The Mycobacterium tuberculosis mmpL7 gene, encoding a hypothetical resistance nodulation division transporter, confers a high resistance level to isoniazid when overexpressed in Mycobacterium smegmatis. The resistance level decreased in the presence of the efflux pump inhibitors reserpine and CCCP (carbonyl cyanide m chlorophenylhydrazone). Energy-dependent efflux of isoniazid from M. smegmatis cells expressing the mmpL7 gene was observed. PMID- 16251329 TI - Differential antibiotic susceptibilities of starved Mycobacterium tuberculosis isolates. AB - We compared the efficacies of 17 agents against rapidly growing and starved Mycobacterium tuberculosis H37Rv. Few compounds have significant activity at attainable concentrations. However, two phenothiazine compounds at similar concentrations were bactericidal for starved and growing cells. These drugs appear to target a process important in both replicating and nonreplicating bacteria. PMID- 16251330 TI - Use of an efflux-deficient streptococcus pneumoniae strain panel to identify ABC class multidrug transporters involved in intrinsic resistance to antimicrobial agents. AB - Thirteen derivatives of the Streptococcus pneumoniae TIGR4 strain in which putative drug efflux pumps were genetically inactivated were constructed and characterized. The results indicate that two linked genes encoding the ABC-type transporters SP2073 and SP2075 function together to confer intrinsic resistance to a series of structurally unrelated compounds, including certain fluoroquinolones. PMID- 16251331 TI - High prevalence of glycopeptide resistance genes vanB, vanD, and vanG not associated with enterococci in human fecal flora. AB - The presence of Enterococcus-associated vancomycin resistance genes vanA, vanB, vanD, vanE, and vanG in rectal swabs was investigated in two hospitals using PCR. All vanA genes detected were associated with vancomycin-resistant enterococci (VRE), whereas VRE-associated vanB genes were detected in only one hospital (4.7%). However, in both hospitals, high prevalences of vanB (6.2 and 2.3%), vanD (43.8 and 26.7%), and vanG (10.5 and 6.9%) genes not associated with enterococci were found. PMID- 16251332 TI - Efficacy of penicillin versus cefdinir in eradication of group A streptococci and tonsillar flora. AB - Core tonsillar cultures were obtained from 40 children with recurrent tonsillitis treated with either penicillin or cefdinir. Group A beta-hemolytic streptococci were isolated from 11 penicillin- and 3 cefdinir-treated (P < 0.001) patients. beta-Lactamase producers were recovered from 17 penicillin- and 3 cefdinir treated (P < 0.01) patients. Inhibiting alpha-hemolytic streptococci were isolated less often from penicillin-treated patients than from cefdinir-treated patients. PMID- 16251333 TI - Therapeutic effects of bacteriophage Cpl-1 lysin against Streptococcus pneumoniae endocarditis in rats. AB - Cpl-1, a pneumococcal phage lytic enzyme, was tested in rats with experimental endocarditis due to Streptococcus pneumoniae WB4. High-dose regimen Cpl-1 eliminated pneumococci from blood within 30 min and decreased bacterial titers in vegetations (>4 log10 CFU/g) within 2 h. Rapid bacterial lysis induced by Cpl-1 treatment increased cytokine secretion noticeably. PMID- 16251334 TI - Recovery of interfering bacteria in the nasopharynx following antimicrobial therapy of acute maxillary sinusitis with telithromycin or amoxicillin clavulanate. AB - The effect on the nasopharyngeal flora of 7 days of amoxicillin-clavulanate or 5 days of 800 mg once a day was studied in 50 adults with acute sinusitis. The numbers of potential pathogens and interfering alpha-hemolytic streptococci were equally reduced after both therapies. However, the number of interfering Prevotella spp. declined more significantly after amoxicillin-clavulanate treatment. PMID- 16251335 TI - Effectiveness of anidulafungin in eradicating Candida species in invasive candidiasis. AB - In a phase 2 open-label, dose-ranging study in patients with candidemia, anidulafungin was effective in eradicating Candida albicans and other species of Candida. The anidulafungin MIC distribution showed that Candida albicans and C. glabrata were the most susceptible species and C. parapsilosis was the least susceptible species. PMID- 16251336 TI - Multiplex PCR for simultaneous detection of macrolide and tetracycline resistance determinants in streptococci. AB - Resistance to macrolides and tetracyclines is increasing among streptococci and co-occurs as their resistance determinants are carried on the same mobile element. We developed a multiplex PCR to facilitate simultaneous and specific detection of resistance determinants for both macrolides [erm(A), erm(B), and mef(A/E)] and tetracyclines [tet(M), tet(O), tet(K), and tet(L)] in streptococci. PMID- 16251337 TI - New Mycobacterium avium antifolate shows synergistic effect when used in combination with dihydropteroate synthase inhibitors. AB - Mycobacterium avium complex (MAC) is resistant to trimethoprim, an inhibitor of bacterial dihydrofolate reductase (DHFR). A previously identified selective inhibitor of MAC DHFR, SRI-8858, was shown to have synergistic activity in combination with dapsone and sulfamethoxazole, two drugs that inhibit bacterial dihydropteroate synthase. PMID- 16251338 TI - Impact of mass distribution of azithromycin on the antibiotic susceptibilities of ocular Chlamydia trachomatis. AB - In a community of Tanzania where trachoma is endemic, we cultured conjunctival swabs from all residents who had active trachoma and were PCR positive for ocular Chlamydia trachomatis, both before (43 isolates) and 2 months after (9 isolates) mass antibiotic treatment. No clinically or programmatically significant increase in azithromycin or tetracycline resistance was observed. PMID- 16251339 TI - Mutations conferring drug resistance in malaria parasite drug transporters Pgh1 and PfCRT do not affect steady-state vacuolar Ca2+. PMID- 16251340 TI - First outbreak of Klebsiella pneumoniae clinical isolates producing GES-5 and SHV 12 extended-spectrum beta-lactamases in Korea. PMID- 16251341 TI - Clinical Isolates of the Spain14-5 clone of Streptococcus pneumoniae carry a recombinant rpoB gene. PMID- 16251342 TI - Pharmacokinetics of colistin methanesulfonate and colistin in a critically ill patient receiving continuous venovenous hemodiafiltration. PMID- 16251343 TI - Improved outcomes with linezolid for methicillin-resistant Staphylococcus aureus infections: better drug or reduced vancomycin susceptibility? PMID- 16251346 TI - Cortical recruitment and nuclear-cytoplasmic shuttling of Scd5p, a protein phosphatase-1-targeting protein involved in actin organization and endocytosis. AB - Scd5p regulates endocytosis and cortical actin organization as a targeting subunit for the Ser/Thr protein phosphatase-1 (PP1) in yeast. To identify localization signals in Scd5p required for cell surface recruitment, visualization of GFP-tagged Scd5 truncations and deletions was performed. Scd5p contains a PP1 binding site, a 3-repeat region of 20 amino acids (3R), and a 9 repeat region of 12 amino acids (9R). We found that the 9R is critical for cortical localization of Scd5p, but cortical recruitment is not essential for Scd5p's function in actin organization and endocytosis. We propose that Scd5p can target PP1 to endocytic factors in the cytoplasm that have been disassembled and/or inactivated by phosphorylation. We also found that Scd5p undergoes nuclear cytoplasmic shuttling in a Crm1p-dependent manner. Scd5p-DeltaCT lacking the 9R region and its nuclear export signal (NES) accumulates in the nucleus, causing cortical actin and endocytic defects. Cytoplasmic localization and function of Scd5p-DeltaCT is restored by NES addition. However, removal of Scd5p's nuclear localization signal prevents nuclear entry, but endocytosis and actin organization remain relatively normal. These results indicate that nuclear cytoplasmic shuttling is not required for regulation of Scd5p's cortical function and suggest that Scd5p has an independent nuclear function. PMID- 16251347 TI - Type I collagen structure regulates cell morphology and EGF signaling in primary rat hepatocytes through cAMP-dependent protein kinase A. AB - Adhesion to type 1 collagen elicits different responses dependent on whether the collagen is in fibrillar (gel) or monomeric form (film). Hepatocytes adherent to collagen film spread and proliferate, whereas those adherent to collagen gel remain rounded and growth arrested. To explore the role of potential intracellular inhibitory signals responsible for collagen gel-mediated growth arrest, cAMP-dependent protein kinase A (PKA) was examined in hepatocytes adherent to collagen film or gel. PKA activity was higher in hepatocytes on collagen gel than on film during G1 of the hepatocyte cell cycle. Inhibition of PKA using H89 increased cell spreading on collagen gel in an EGF-dependent manner, whereas activation of PKA using 8-Br-cAMP decreased cell spreading on collagen film. PKA inhibition also restored ERK activation, cyclin D1 expression and G1-S progression on collagen gel, but had no effect on cells adherent to collagen film. Analysis of EGF receptor phosphorylation revealed that adhesion to collagen gel alters tyrosine phosphorylation of the EGF receptor, leading to reduced phosphorylation of tyrosine residue 845, which was increased by inhibition of PKA. These results demonstrate that fibrillar type 1 collagen can actively disrupt cell cycle progression by inhibiting specific signals from the EGF receptor through a PKA-dependent pathway. PMID- 16251350 TI - Unique suppression of prostaglandin H synthase-2 expression by inhibition of histone deacetylation, specifically in human amnion but not adjacent choriodecidua. AB - The key molecular regulatory mechanisms that govern and coordinate the molecular alterations that underpin the process of human labor remain incompletely understood although enhanced intrauterine prostaglandin production is known to be requisite. Studies from cancer tissues have indicated that at least one key enzyme of prostaglandin biosynthesis can have its activity severely reduced by increased histone deacetylation and enhanced DNA methylation status. We have advanced the hypothesis that similar regulation may occur in intrauterine tissues during pregnancy to prevent inadvertent activation of this powerful initiating signal by dampening responses to premature activation by agents such as cytokines. Our studies have shown that responsiveness of amnion, a key intrauterine tissue, to interleukin-1beta is abrogated by inhibition of histone deacetylation, whereas PGDH amounts were increased basally. The findings do integrate well with others concerning progesterone (inhibitory) actions such that a decrease in the level of histone acetylation in human gestational tissues near term might herald a coordinated series of events that all result in a positive drive for parturition. Hence, a new level of regulatory action and potential therapeutic targets for pathologies such as preterm labor can flow from these findings. PMID- 16251348 TI - The homologous putative GTPases Grn1p from fission yeast and the human GNL3L are required for growth and play a role in processing of nucleolar pre-rRNA. AB - Grn1p from fission yeast and GNL3L from human cells, two putative GTPases from the novel HSR1_MMR1 GTP-binding protein subfamily with circularly permuted G motifs play a critical role in maintaining normal cell growth. Deletion of Grn1 resulted in a severe growth defect, a marked reduction in mature rRNA species with a concomitant accumulation of the 35S pre-rRNA transcript, and failure to export the ribosomal protein Rpl25a from the nucleolus. Deleting any of the Grn1p G-domain motifs resulted in a null phenotype and nuclear/nucleolar localization consistent with the lack of nucleolar export of preribosomes accompanied by a distortion of nucleolar structure. Heterologous expression of GNL3L in a Deltagrn1 mutant restored processing of 35S pre-rRNA, nuclear export of Rpl25a and cell growth to wild-type levels. Genetic complementation in yeast and siRNA knockdown in HeLa cells confirmed the homologous proteins Grn1p and GNL3L are required for growth. Failure of two similar HSR1_MMR1 putative nucleolar GTPases, Nucleostemin (NS), or the dose-dependent response of breast tumor autoantigen NGP 1, to rescue deltagrn1 implied the highly specific roles of Grn1p or GNL3L in nucleolar events. Our analysis uncovers an important role for Grn1p/GNL3L within this unique group of nucleolar GTPases. PMID- 16251349 TI - Mitochondrial AKAP121 links cAMP and src signaling to oxidative metabolism. AB - AKAP121 focuses distinct signaling events from membrane to mitochondria by binding and targeting cAMP-dependent protein kinase (PKA), protein tyrosine phosphatase (PTPD1), and mRNA. We find that AKAP121 also targets src tyrosine kinase to mitochondria via PTPD1. AKAP121 increased src-dependent phosphorylation of mitochondrial substrates and enhanced the activity of cytochrome c oxidase, a component of the mitochondrial respiratory chain. Mitochondrial membrane potential and ATP oxidative synthesis were enhanced by AKAP121 in an src- and PKA dependent manner. Finally, siRNA-mediated silencing of endogenous AKAP121 drastically impaired synthesis and accumulation of mitochondrial ATP. These findings indicate that AKAP121, through its role in enhancing cAMP and tyrosine kinase signaling to distal organelles, is an important regulator in mitochondrial metabolism. PMID- 16251351 TI - Phosphatidylinositol 3-phosphate indirectly activates KCa3.1 via 14 amino acids in the carboxy terminus of KCa3.1. AB - KCa3.1 is an intermediate conductance Ca2+-activated K+ channel that is expressed predominantly in hematopoietic cells, smooth muscle cells, and epithelia where it functions to regulate membrane potential, Ca2+ influx, cell volume, and chloride secretion. We recently found that the KCa3.1 channel also specifically requires phosphatidylinositol-3 phosphate [PI(3)P] for channel activity and is inhibited by myotubularin-related protein 6 (MTMR6), a PI(3)P phosphatase. We now show that PI(3)P indirectly activates KCa3.1. Unlike KCa3.1 channels, the related KCa2.1, KCa2.2, or KCa2.3 channels do not require PI(3)P for activity, suggesting that the KCa3.1 channel has evolved a unique means of regulation that is critical for its biological function. By making chimeric channels between KCa3.1 and KCa2.3, we identified a stretch of 14 amino acids in the carboxy-terminal calmodulin binding domain of KCa3.1 that is sufficient to confer regulation of KCa2.3 by PI(3)P. However, mutation of a single potential phosphorylation site in these 14 amino acids did not affect channel activity. These data together suggest that PI(3)P and these 14 amino acids regulate KCa3.1 channel activity by recruiting an as yet to be defined regulatory subunit that is required for Ca2+ gating of KCa3.1. PMID- 16251352 TI - The CNA1 histone of the ciliate Tetrahymena thermophila is essential for chromosome segregation in the germline micronucleus. AB - Ciliated protozoans present several features of chromosome segregation that are unique among eukaryotes, including their maintenance of two nuclei: a germline micronucleus, which undergoes conventional mitosis and meiosis, and a somatic macronucleus that divides by an amitotic process. To study ciliate chromosome segregation, we have identified the centromeric histone gene in the Tetrahymena thermophila genome (CNA1). CNA1p specifically localizes to peripheral centromeres in the micronucleus but is absent in the macronucleus during vegetative growth. During meiotic prophase of the micronucleus, when chromosomes are stretched to twice the length of the cell, CNA1p is found localized in punctate spots throughout the length of the chromosomes. As conjugation proceeds, CNA1p appears initially diffuse, but quickly reverts to discrete dots in those nuclei destined to become micronuclei, whereas it remains diffuse and is gradually lost in developing macronuclei. In progeny of germline CNA1 knockouts, we see no defects in macronuclear division or viability of the progeny cells immediately following the knockout. However, within a few divisions, progeny show abnormal mitotic segregation of their micronucleus, with most cells eventually losing their micronucleus entirely. This study reveals a strong dependence of the germline micronucleus on centromeric histones for proper chromosome segregation. PMID- 16251353 TI - DNA replication origins fire stochastically in fission yeast. AB - DNA replication initiates at discrete origins along eukaryotic chromosomes. However, in most organisms, origin firing is not efficient; a specific origin will fire in some but not all cell cycles. This observation raises the question of how individual origins are selected to fire and whether origin firing is globally coordinated to ensure an even distribution of replication initiation across the genome. We have addressed these questions by determining the location of firing origins on individual fission yeast DNA molecules using DNA combing. We show that the firing of replication origins is stochastic, leading to a random distribution of replication initiation. Furthermore, origin firing is independent between cell cycles; there is no epigenetic mechanism causing an origin that fires in one cell cycle to preferentially fire in the next. Thus, the fission yeast strategy for the initiation of replication is different from models of eukaryotic replication that propose coordinated origin firing. PMID- 16251354 TI - Rac1 leads to phosphorylation-dependent increase in stability of the p66shc adaptor protein: role in Rac1-induced oxidative stress. AB - The rac1 GTPase and the p66shc adaptor protein regulate intracellular levels of reactive oxygen species (ROS). We examined the relationship between rac1 and p66shc. Expression of constitutively active rac1 (rac1V12) increased phosphorylation, reduced ubiquitination, and increased stability of p66shc protein. Rac1V12-induced phosphorylation and up-regulation of p66shc was suppressed by inhibiting p38MAPK and was dependent on serine 54 and threonine 386 in p66shc. Phosphorylation of recombinant p66shc by p38MAPK in vitro was also partly dependent on serine 54 and threonine 386. Reconstitution of p66shc in p66shc-null fibroblasts increased intracellular ROS generated by rac1V12, which was significantly dependent on the integrity of residues 54 and 386. Overexpression of p66shc increased rac1V12-induced apoptosis, an effect that was also partly dependent on serine 54 and threonine 386. Finally, RNA interference mediated down-regulation of endogenous p66shc suppressed rac1V12-induced cell death. These findings identify p66shc as a mediator of rac1-induced oxidative stress. In addition, they suggest that serine 54 and threonine 386 are novel phosphorylatable residues in p66shc that govern rac1-induced increase in its expression, through a decrease in its ubiquitination and degradation, and thereby mediate rac1-stimulated cellular oxidative stress and death. PMID- 16251355 TI - The thioredoxin system protects ribosomes against stress-induced aggregation. AB - We previously showed that thioredoxins are required for dithiothreitol (DTT) tolerance, suggesting they maintain redox homeostasis in response to both oxidative and reductive stress conditions. In this present study, we screened the complete set of viable deletion strains in Saccharomyces cerevisiae for sensitivity to DTT to identify cell functions involved in resistance to reductive stress. We identified 195 mutants, whose gene products are localized throughout the cell. DTT-sensitive mutants were distributed among most major biological processes, but they particularly affected gene expression, metabolism, and the secretory pathway. Strikingly, a mutant lacking TSA1, encoding a peroxiredoxin, showed a similar sensitivity to DTT as a thioredoxin mutant. Epistasis analysis indicated that thioredoxins function upstream of Tsa1 in providing tolerance to DTT. Our data show that the chaperone function of Tsa1, rather than its peroxidase function, is required for this activity. Cells lacking TSA1 were found to accumulate aggregated proteins, and this was exacerbated by exposure to DTT. Analysis of the protein aggregates revealed that they are predominantly composed of ribosomal proteins. Furthermore, aggregation was found to correlate with an inhibition of translation initiation. We propose that Tsa1 normally functions to chaperone misassembled ribosomal proteins, preventing the toxicity that arises from their aggregation. PMID- 16251356 TI - A genomewide screen reveals a role of mitochondria in anaerobic uptake of sterols in yeast. AB - The mechanisms that govern intracellular transport of sterols in eukaryotic cells are not well understood. Saccharomyces cerevisiae is a facultative anaerobic organism that becomes auxotroph for sterols and unsaturated fatty acids in the absence of oxygen. To identify pathways that are required for uptake and transport of sterols, we performed a systematic screen of the yeast deletion mutant collection for genes that are required for growth under anaerobic conditions. Of the approximately 4800 nonessential genes represented in the deletion collection, 37 were essential for growth under anaerobic conditions. These affect a wide range of cellular functions, including biosynthetic pathways for certain amino acids and cofactors, reprogramming of transcription and translation, mitochondrial function and biogenesis, and membrane trafficking. Thirty-three of these mutants failed to grow on lipid-supplemented media when combined with a mutation in HEM1, which mimics anaerobic conditions in the presence of oxygen. Uptake assays with radio- and fluorescently labeled cholesterol revealed that 17 of the 33 mutants strongly affect uptake and/or esterification of exogenously supplied cholesterol. Examination of the subcellular distribution of sterols in these uptake mutants by cell fractionation and fluorescence microscopy indicates that some of the mutants block incorporation of cholesterol into the plasma membrane, a presumably early step in sterol uptake. Unexpectedly, the largest class of uptake mutants is affected in mitochondrial functions, and many of the uptake mutants show electron-dense mitochondrial inclusions. These results indicate that a hitherto uncharacterized mitochondrial function is required for sterol uptake and/or transport under anaerobic conditions and are discussed in light of the fact that mitochondrial import of cholesterol is required for steroidogenesis in vertebrate cells. PMID- 16251357 TI - Essential roles in development and pigmentation for the Drosophila copper transporter DmATP7. AB - Defects in the mammalian Menkes and Wilson copper transporting P-type ATPases cause severe copper homeostasis disease phenotypes in humans. Here, we find that DmATP7, the sole Drosophila orthologue of the Menkes and Wilson genes, is vital for uptake of copper in vivo. Analysis of a DmATP7 loss-of-function allele shows that DmATP7 is essential in embryogenesis, early larval development, and adult pigmentation and is probably required for copper uptake from the diet. These phenotypes are analogous to those caused by mutation in the mouse and human Menkes genes, suggesting that like Menkes, DmATP7 plays at least two roles at the cellular level: delivering copper to cuproenzymes required for pigmentation and neuronal function and removing excess cellular copper via facilitated efflux. DmATP7 displays a dynamic and unexpected expression pattern in the developing embryo, implying novel functions for this copper pump and the lethality observed in DmATP7 mutant flies is the earliest seen for any copper homeostasis gene. PMID- 16251358 TI - Interactions between EHD proteins and Rab11-FIP2: a role for EHD3 in early endosomal transport. AB - Eps15 homology domain (EHD) 1 enables membrane recycling by controlling the exit of internalized molecules from the endocytic recycling compartment (ERC) en route to the plasma membrane, similar to the role described for Rab11. However, no physical or functional connection between Rab11 and EHD-family proteins has been demonstrated yet, and the mode by which they coordinate their regulatory activity remains unknown. Here, we demonstrate that EHD1 and EHD3 (the closest EHD1 paralog), bind to the Rab11-effector Rab11-FIP2 via EH-NPF interactions. The EHD/Rab11-FIP2 associations are affected by the ability of the EHD proteins to bind nucleotides, and Rab11-FIP2 is recruited to EHD-containing membranes. These results are consistent with a coordinated role for EHD1 and Rab11-FIP2 in regulating exit from the ERC. However, because no function has been attributed to EHD3, the significance of its interaction with Rab11-FIP2 remained unclear. Surprisingly, loss of EHD3 expression prevented the delivery of internalized transferrin and early endosomal proteins to the ERC, an effect differing from that described upon EHD1 knockdown. Moreover, the subcellular localization of Rab11-FIP2 and endogenous Rab11 were altered upon EHD3 knockdown, with both proteins absent from the ERC and retained in the cell periphery. The results presented herein promote a coordinated role for EHD proteins and Rab11-FIP2 in mediating endocytic recycling and provide evidence for the function of EHD3 in early endosome to ERC transport. PMID- 16251360 TI - Actin is required for endocytosis at the apical surface of Madin-Darby canine kidney cells where ARF6 and clathrin regulate the actin cytoskeleton. AB - In epithelial cell lines, apical but not basolateral clathrin-mediated endocytosis has been shown to be affected by actin-disrupting drugs. Using electron and fluorescence microscopy, as well as biochemical assays, we show that the amount of actin dedicated to endocytosis is limiting at the apical surface of epithelia. In part, this contributes to the low basal rate of clathrin-dependent endocytosis observed at this epithelial surface. ARF6 in its GTP-bound state triggers the recruitment of actin from the cell cortex to the clathrin-coated pit to enable dynamin-dependent endocytosis. In addition, we show that perturbation of the apical endocytic system by expression of a clathrin heavy-chain mutant results in the collapse of microvilli. This phenotype was completely reversed by the expression of an ARF6-GTP-locked mutant. These observations indicate that concomitant to actin recruitment, the apical clathrin endocytic system is deeply involved in the morphology of the apical plasma membrane. PMID- 16251359 TI - Cis-Golgi matrix proteins move directly to endoplasmic reticulum exit sites by association with tubules. AB - The role of cis-medial Golgi matrix proteins in retrograde traffic is poorly understood. We have used imaging techniques to understand the relationship between the cis-medial Golgi matrix and transmembrane proteins during retrograde traffic in control and brefeldin A (BFA)-treated cells. All five of the cis medial matrix proteins tested were associated with retrograde tubules within 2-3 min of initiation of tubule formation. Then, at later time points (3-10 min), transmembrane proteins are apparent in the same tubules. Strikingly, both the matrix proteins and the transmembrane proteins moved directly to endoplasmic reticulum (ER) exit sites labeled with p58 and Sec13, and there seemed to be a specific interaction between the ER exit sites and the tips or branch points of the tubules enriched for the matrix proteins. After the initial interaction, Golgi matrix proteins accumulated rapidly (5-10 min) at ER exit sites, and Golgi transmembrane proteins accumulated at the same sites approximately 2 h later. Our data suggest that Golgi cis-medial matrix proteins participate in Golgi-to-ER traffic and play a novel role in tubule formation and targeting. PMID- 16251361 TI - Proteolytic cleavage and phosphorylation of a tumor-associated ErbB4 isoform promote ligand-independent survival and cancer cell growth. AB - The ErbB1 and ErbB2 receptors are oncogenes with therapeutic significance in human cancer, whereas the transforming potential of the related ErbB4 receptor has remained controversial. Here, we have addressed whether four alternatively spliced ErbB4 isoforms differ in regulating cellular responses relevant for tumor growth. We show that the two tumor necrosis factor-alpha converting enzyme (TACE) cleavable ErbB4 isoforms (the juxtamembrane [JM]-a isoforms) were overexpressed in a subset of primary human breast cancers together with TACE. The overexpression of the JM-a cytoplasmic (CYT)-2 ErbB4 isoform promoted ErbB4 phosphorylation, survival of interleukin-3-dependent cells, and proliferation of breast cancer cells even in the absence of ligand stimulation, whereas activation of the other three ErbB4 isoforms required ligand stimulation. Ligand-independent cellular responses to ErbB4 JM-a CYT-2 overexpression were regulated by both tyrosine kinase activity and a two-step proteolytic generation of an intracellular receptor fragment involving first a TACE-like proteinase, followed by gamma-secretase activity. These data suggest a novel transforming mechanism for the ErbB4 receptor in human breast cancer that is 1) specific for a single receptor isoform and 2) depends on proteinase cleavage and kinase activity but not ligand activation of the receptor. PMID- 16251362 TI - Visualization of vacuolar acidification-induced transcription of genes of pathogens inside macrophages. AB - The objective of these studies was to analyze the role of the ionic environment of phagosomal vacuoles in the control of pathogens by macrophages. Digital imaging and flow cytometry were used to follow the induction of the phoP promoter of Salmonella enterica Typhimurium within live macrophages. Manipulating the Mg2+ concentration within the Salmonella-containing vacuole (SCV) was without effect on the early induction of PhoPQ. Moreover, direct measurement of [Mg2+] within the SCV using nanosensor particles showed that, during this initial period of phoP activation, the concentration of the divalent cation is rapidly regulated and stabilizes around 1 mm. Extrusion of other divalent cations via the Nramp1 efflux pump was similarly ruled out as an important contributor to the activation of the regulon. By contrast, induction of PhoP was greatly attenuated when the pH gradient across the SCV membrane was dissipated. A second, more modest pH independent component of PhoP induction was unmasked by inhibition of the vacuolar proton pump. This second component was eliminated by pretreatment of cells with IFNgamma, even though the cytokine augmented the overall PhoP response. These findings demonstrate the existence of at least three separate activators of phoP transcription: resting and IFNgamma-stimulated pH-sensitive components, plus a pH-independent component. PMID- 16251363 TI - Undistorted structural analysis of soluble proteins by attenuated total reflectance infrared spectroscopy. AB - Water from the solvent very strongly absorbs light in the frequency range of interest for studying protein structure by infrared (IR) spectroscopy. This renders handling of the observation cells painstaking and time consuming, and limits the reproducibility of the measurements when IR spectroscopy is applied to proteins in aqueous solutions. These difficulties are circumvented by the use of an Attenuated Total Reflectance (ATR) accessory. However, when protein solutions are studied, ATR spectroscopy suffers from several drawbacks, the most severe being nonproportionality of the observed absorbance with the protein concentration and spectral distortions that vary from protein to protein and from sample to sample. In this study, we show (1) that the nonproportionality is due to adsorption of the protein on the ATR crystal surface; (2) that the contribution of the crystal-adsorbed protein can easily be taken into account, rendering the corrected absorbance proportional to the protein concentration; (3) that the observed variable base line distortions, likely due to changes in the penetration depth of the light beam in solutions with the refractive index that depends on the protein concentration, can be easily eliminated; and (4) that ATR IR spectra thus corrected for protein adsorption and light penetration can be used to properly analyze the secondary structure of proteins in solution. PMID- 16251364 TI - A novel representation of protein sequences for prediction of subcellular location using support vector machines. AB - As the number of complete genomes rapidly increases, accurate methods to automatically predict the subcellular location of proteins are increasingly useful to help their functional annotation. In order to improve the predictive accuracy of the many prediction methods developed to date, a novel representation of protein sequences is proposed. This representation involves local compositions of amino acids and twin amino acids, and local frequencies of distance between successive (basic, hydrophobic, and other) amino acids. For calculating the local features, each sequence is split into three parts: N-terminal, middle, and C terminal. The N-terminal part is further divided into four regions to consider ambiguity in the length and position of signal sequences. We tested this representation with support vector machines on two data sets extracted from the SWISS-PROT database. Through fivefold cross-validation tests, overall accuracies of more than 87% and 91% were obtained for eukaryotic and prokaryotic proteins, respectively. It is concluded that considering the respective features in the N terminal, middle, and C-terminal parts is helpful to predict the subcellular location. PMID- 16251365 TI - Insights into the role of the metal binding site in methionine-R-sulfoxide reductases B. AB - Methionine sulfoxide reductases B (MsrBs) catalyze the reduction of methionine-R sulfoxide via a three-step chemical mechanism including a reductase step, formation of an intradisulfide bond followed by a thioredoxin recycling process. Fifty percent of the MsrBs, including the Escherichia coli enzyme, possess a metal binding site composed of two CXXC motifs of unknown function. It is located on the opposite side of the active site. The overexpressed E. coli MsrB tightly binds one atom of zinc/iron. Substitution of the cysteines of E. coli MsrB results in complete loss of bound metal and reductase activity, and leads to a low-structured conformation of the protein as shown by CD, fluorescence, and DSC experiments. Introduction of the two CXXC motifs in Neisseria meningitidis MsrB domain leads to a MsrB that tightly binds one atom of zinc/iron, shows a strongly increased thermal stability and displays a reductase activity similar to that of the wild-type but lacking thioredoxin recycling activity. These results demonstrate the stabilizing effect of the metal and the existence of a preformed metal binding site in the nonbound metal MsrB. The data also indicate that metal binding to N. meningitidis MsrB induces subtle structural modifications, which prevent formation of a competent binary complex between oxidized MsrB and reduced thioredoxin but not between reduced MsrB and substrate. The fact that the E. coli and the N. meningitidis MsrBs exhibit a similar thermal stability suggests the existence of other structural factors in the nonbound metal MsrBs that compensate the metal bound stabilizing effect. PMID- 16251367 TI - Hydrogen-bonded turns in proteins: the case for a recount. AB - Beta-turns are sites at which proteins change their overall chain direction, and they occur with high frequency in globular proteins. The Protein Data Bank has many instances of conformations that resemble beta-turns but lack the characteristic N-H(i) --> O=C(i - 3) hydrogen bond of an authentic beta-turn. Here, we identify potential hydrogen-bonded beta-turns in the coil library, a Web accessible database utility comprised of all residues not in repetitive secondary structure, neither alpha-helix nor beta-sheet (http://www.roselab.jhu.edu/coil). In particular, candidate turns were identified as four-residue segments satisfying highly relaxed geometric criteria but lacking a strictly defined hydrogen bond. Such candidates were then subjected to a minimization protocol to determine whether slight changes in torsion angles are sufficient to shift the conformation into reference-quality geometry without deviating significantly from the original structure. This approach of applying constrained minimization to known structures reveals a substantial population of previously unidentified, stringently defined, hydrogen-bonded beta-turns. In particular, 33% of coil library residues were classified as beta-turns prior to minimization. After minimization, 45% of such residues could be classified as beta-turns, with another 8% in 3(10) helixes (which closely resemble type III beta-turns). Of the remaining coil library residues, 37% have backbone dihedral angles in left-handed polyproline II structure. PMID- 16251368 TI - Women's occupational health. PMID- 16251369 TI - Mortality and cancer incidence in Ontario glass fiber workers. AB - BACKGROUND: In a previous cohort study of glass fiber manufacturing, we found a significant increase in lung cancer. This study extends the follow-up period. METHODS: We conducted a historical prospective study of 2557 men employed in producing glass wool. We obtained work histories, causes and dates of death, and date and site of cancer diagnoses. We computed standardized mortality and incidence ratios (SMR, SIR). RESULTS: The overall SMR for lung cancer was 163 (95% CI = 118-221). The SMR did not vary consistently by duration of employment and time since first employment. However, plant workers with >20 years' employment and >40 years since first exposure had an SMR for lung cancer of 282 (95% CI = 113-582). The SMR dropped with later date of first exposure, but the trend was non-significant. There was an unexpected overall increase in kidney cancer incidence. DISCUSSION: The increase in lung cancer is greater than in other cohorts of glass fiber workers. Since exposure data are lacking from the early years of the plant, we cannot state if the excess was due to glass fibers, other work exposures or other reasons. PMID- 16251366 TI - Solution structure of Archaeglobus fulgidis peptidyl-tRNA hydrolase (Pth2) provides evidence for an extensive conserved family of Pth2 enzymes in archea, bacteria, and eukaryotes. AB - The solution structure of protein AF2095 from the thermophilic archaea Archaeglobus fulgidis, a 123-residue (13.6-kDa) protein, has been determined by NMR methods. The structure of AF2095 is comprised of four alpha-helices and a mixed beta-sheet consisting of four parallel and anti-parallel beta-strands, where the alpha-helices sandwich the beta-sheet. Sequence and structural comparison of AF2095 with proteins from Homo sapiens, Methanocaldococcus jannaschii, and Sulfolobus solfataricus reveals that AF2095 is a peptidyl-tRNA hydrolase (Pth2). This structural comparison also identifies putative catalytic residues and a tRNA interaction region for AF2095. The structure of AF2095 is also similar to the structure of protein TA0108 from archaea Thermoplasma acidophilum, which is deposited in the Protein Data Bank but not functionally annotated. The NMR structure of AF2095 has been further leveraged to obtain good quality structural models for 55 other proteins. Although earlier studies have proposed that the Pth2 protein family is restricted to archeal and eukaryotic organisms, the similarity of the AF2095 structure to human Pth2, the conservation of key active-site residues, and the good quality of the resulting homology models demonstrate a large family of homologous Pth2 proteins that are conserved in eukaryotic, archaeal, and bacterial organisms, providing novel insights in the evolution of the Pth and Pth2 enzyme families. PMID- 16251370 TI - Mortality of UK electricity generation and transmission workers, 1973-2002. AB - OBJECTIVE: To examine mortality from cancer and non-malignant causes among a large cohort of UK electricity generation and transmission workers. METHODS: The mortality experienced by a cohort of 83,923 employees of the former Central Electricity Generating Board of England and Wales was investigated for the period 1973-2002. All employees had worked for at least 6 months with some employment between 1973 and 1982. Standardized mortality ratios (SMRs) were used to assess mortality in the total cohort and in three sub-cohorts: power station workers, substation and transmission workers and workers at non-operational locations. These classifications were based on the place of work of the first known job. RESULTS: Overall mortality was significantly below that expected, based on national rates [males: observed (Obs) 18,773, expected (Exp) 22,497.9, SMR 83; females: Obs 1122, Exp 1424.9, SMR 79]. Statistically significant deficits of deaths were also found for most of the major disease groupings. However, significant excesses of deaths were found in male power station workers for cancer of the pleura (Obs 129, Exp 30.3, SMR 426) and in male workers from non operational locations for cancer of the brain (Obs 55, Exp 36.0, SMR 153). There was also a non-significant excess of deaths from cancer of the breast in male power station workers (Obs 10, Exp 5.3, SMR 190). CONCLUSIONS: Mortality was exceptionally low for most causes of death but late health effects from earlier asbestos exposure were still in evidence. PMID- 16251371 TI - Does having an occupational mental health service make any difference? AB - BACKGROUND: The effects of mental disorder on UK's workforce are increasing, yet most occupational health (OH) providers have limited expertise in dealing with mental health issues. AIMS: To examine the effectiveness in terms of organizational and clinical outcomes, of an OH liaison psychiatry service in an inner city area. METHODS: A retrospective survey of case notes from the first 2 years of an OH liaison psychiatry service was carried out. RESULTS: Seventy-six cases were identified and 68% were seen within 1 month of referral. After assessment, females were significantly more likely to be given a formal psychiatric diagnosis than males. Overall, 45% of patients had mood disorders, 14% anxiety disorders and two cases of psychosis were identified. Advice was offered to the referring OH practitioner in 80% of cases. CONCLUSIONS: This study found that liaison psychiatrists were able to fulfil a useful role in an OH department. Most of the cases seen within the department would not be classed as serious mental illness and therefore it is unlikely that community mental health teams would become involved in their routine care. Specialist mental health support from an 'in house' service is likely to be of considerable benefit from both occupational and medical perspectives. PMID- 16251372 TI - Prevalence, management and impact of urinary incontinence in the workplace. AB - BACKGROUND: The few prior studies of urinary symptoms in the workplace have been small investigations of women in specific occupations (e.g. nursing) or industries (e.g. pottery manufacture). In this study, the aims were to describe the prevalence, management and impact of urinary incontinence for a large cross section of employed women in the USA. METHODS: Five-page questionnaires were mailed to 5130 American households selected from the National Family Opinion survey panel during the spring of 2004. Usable questionnaires were returned by 3364 women in the target age range of 18-60 years. RESULTS: About 37% of the 2326 employed respondents reported urine loss during the last 30 days. The most common strategies for managing incontinence at work included frequent bathroom breaks and wearing pads. The use of urine control methods increased with the severity of urine loss. The effect of incontinence on workplace activities also increased with the severity of urine loss: 88% of employed women with the most severe symptoms reported at least some negative impact on concentration, performance of physical activities, self-confidence or the ability to complete tasks without interruption. CONCLUSION: Urinary incontinence is prevalent among employed women. Those who experience severe symptoms report that it has a negative effect on aspects of work. Programs on the prevalence and impact of urinary incontinence would help educate both employers and employees, and may lead to the development of better management techniques in the workplace. PMID- 16251373 TI - Health-related quality of life and sickness absence in community nursing home employees: randomized controlled trial of physical exercise. AB - BACKGROUND: It is a common belief that physical exercise at the workplace decreases subjective health complaints and reduces sickness absence, but this is not supported by previous randomized studies. AIMS: To evaluate the effectiveness of physical exercise at the workplace. METHODS: One hundred and twenty-nine employees in a community-based nursing home for the elderly were randomized into physical exercise or control groups. A weekly exercise class consisting of light aerobic exercise, muscle strengthening and stretching was held for a 6-month period. The control group was told to continue their ordinary activity. The main outcome measures were aerobic fitness (UKK, walking test), health-related quality of life (COOP/WONCA) and sickness absence. Blinded assessments were carried out at baseline and following the 6-month intervention. Complete sickness absence data were collected from a community register for two comparable 7-month periods. RESULTS: The average number of exercise sessions was 12 (0-26). Self-reported physical activity increased in the intervention group compared with the control group (P < 0.01). Aerobic fitness improved in both groups (P < 0.01). Mean sickness absence increased from 6.8 to 15.6 days in the exercise group and from 10.4 to 14.5 in the control group. No differences between groups were found for aerobic fitness, health-related quality of life or sickness absence. CONCLUSION: The intervention neither improved health-related quality of life nor reduced sickness absence. PMID- 16251374 TI - From Jenner to modern smallpox vaccines. PMID- 16251375 TI - Risk factors for cytomegalovirus seropositivity in a population of day care educators in Montreal, Canada. AB - BACKGROUND: Horizontal transmission of cytomegalovirus (CMV) is common in the day care setting. Day care educators appear to be at a high risk of infection; however, studies are limited. AIMS: To determine the proportion of CMV seropositive female educators in the day care setting and to identify associated risk factors. METHODS: Educator- and day care-level risk factors for CMV seropositivity were obtained by questionnaires from day care educators and directors, respectively. Sera were collected from educators and analyzed by enzyme-linked immunosorbent assay. Significant independent risk factors for CMV seropositivity were determined using a multivariable logistic regression model which was fitted using the generalized estimating equation method. RESULTS: CMV seroprevalence in 473 female educators from 81 day care centers in Montreal, Canada, was 57%. Significant risk factors for CMV seropositivity were (i) increasing age (OR5-yr = 1.19; 95% CI = 1.05-1.35), (ii) low-income country of birth (OR = 10.23; 95% CI = 2.64-39.50) or middle-income country of birth (OR = 4.99; 95% CI = 2.39-10.40), (iii) having > or =2 children of their own (OR = 1.98; 95% CI = 1.19-3.31) and (iv) child-to-educator ratio >6 (18-35 months old) in a day care center (OR = 1.87; 95% CI = 1.25-2.81). CONCLUSIONS: Day care educators have risks for CMV infection related to their work in the day care setting, as well as personal risk factors. A review of current guidelines for the prevention of CMV infection in day care is needed to ensure that recommendations are evidence based. PMID- 16251376 TI - Psychosocial work conditions, unemployment and self-reported psychological health: a population-based study. AB - AIM: To investigate the association between psychosocial conditions at work, unemployment and self-reported psychological health. METHODS: A cross-sectional postal questionnaire for the 2000 public health survey in Scania was administered to both working and unemployed people aged 18-64 years. Logistic regression models were used to investigate the association between psychosocial factors at work/unemployment and self-reported psychological health (General Health Questionnaire 12). Psychosocial conditions at work were classified according to the Karasek-Theorell demand-control/decision latitudes into relaxed, active, passive and job strain. The multivariate analyses included age, country of origin, education, economic stress and social participation. RESULTS: A total of 5180 people returned their questionnaire, giving a participation rate of 59%. Fifteen per cent of men and 20% of women reported poor psychological health. Those with high demands and high control (active category), those with high demands and low control (job strain category) and the unemployed had significantly higher odds ratios of poor psychological health compared to those with low demands and high control (relaxed category). Those with low demands and low control (passive category) did not differ significantly from the relaxed category. The associations remained in the multivariate analyses. CONCLUSIONS: The study found that certain psychosocial work factors are associated with higher levels of self-reported psychological ill-health and illustrates the great importance of psychosocial conditions in determining psychological health at the population level. As found elsewhere, being unemployed was an even stronger predictor of psychological ill-health. PMID- 16251377 TI - Glutaraldehyde-induced occupational asthma: BALF components and BALF and serum Clara cell protein (CC16) changes due to specific inhalatory provocation test. AB - OBJECTIVES: The purpose of this study was to evaluate bronchoalveolar lavage fluid (BALF) components and Clara cell protein (CC16) concentration in serum and BALF in patients with glutaraldehyde (GA)-induced asthma, before and after a specific inhalatory provocation test (SIPT) with GA, in comparison to atopic asthmatics and healthy individuals. METHODS: Spirometry and bronchoalveolar lavage were performed before and after SIPT. The serum and BALF concentrations of CC16 and cytogram content in BALF were evaluated. RESULTS: In GA-sensitized asthmatics, the level of CC16 in BALF and serum was significantly lower at 24 h after SIPT in comparison with the values recorded prior to the experiment. There was a significant increase in the proportion of eosinophils, basophils and lymphocytes in BALF of GA-sensitized asthmatics obtained after SIPT. CONCLUSIONS: The determination of CC16 either in serum or in BALF is a non-invasive test to detect Clara cell damage. PMID- 16251378 TI - Evidence-based decision making in an endoscopy nurse with respiratory symptoms exposed to the new ortho-phthalaldehyde (OPA) disinfectant. AB - BACKGROUND: ortho-Phthalaldehyde (OPA) can cause mucous irritation, respiratory symptoms and IgE-mediated hypersensitivity reactions. Very little information is available about OPA-related effects in health personnel. AIM: To report the decision-making process for the case of an endoscopy nurse complaining of cough and burning of the nose and throat during OPA exposure at work. METHODS: The problem focused on the relationship between OPA exposure and the respiratory symptoms and was investigated using an evidence-based (EB) medicine paradigm. RESULTS: A literature search was performed using the database Medline and the search engine Google. Papers and guidelines were assessed for their suitability in the EB case identification of suspected occupational asthma (OA). A multistep approach suggested by a guideline was considered most appropriate for practical use. The nurse shared the decision-making process and underwent evaluation of the clinical suspicion index and interventions for diagnosis of OA. Despite the high clinical suspicion index, the diagnosis of OA was excluded and any work restriction was avoided. Health surveillance follow-up showed a good clinical outcome and prompt recovery from respiratory symptoms after improvement of environmental control measures. CONCLUSION: The case study shows that the implementation of EB guidelines provides the occupational physician with an appropriate decision-making process for the identification and management of workers with suspected OA. Screening out of OA is highly relevant because diagnosis of disease requires removal from exposure and frequently impacts negatively on worker employment. PMID- 16251379 TI - Re: mortality experience of male workers at a UK tin smelter. PMID- 16251380 TI - Latent period for symptomatic sensitization in bakeries. PMID- 16251381 TI - Expression of the UDP-GalNAc: polypeptide N-acetylgalactosaminyltransferase family is spatially and temporally regulated during Drosophila development. AB - The UDP-GalNAc : polypeptide N-acetylgalactosaminyltransferase (ppGaNTase or ppGalNAcT or pgant) enzyme family is responsible for the first committed step in the synthesis of mucin-type O-glycans on protein substrates. Previous work from our group has demonstrated both sequence and functional conservation between members of this family in mammals and the fruit fly, Drosophila melanogaster. One member of this family in Drosophila has been shown to be essential for viability and development. In an effort to understand the developmental stages and processes in which O-glycosylation is involved, we have determined the expression pattern of each functional family member as well as putative isoforms during Drosophila development. Our studies indicate that isoforms are expressed in discrete spatial and temporal fashions during development, with some isoforms being found uniquely in restricted areas of the developing embryo (brain, trachea, pharynx, esophagus, proventriculus, and amnioserosa), whereas others are found in multiple regions and overlap with the expression of other isoforms (salivary glands, posterior midgut, anterior midgut, and the fore-/hindgut) during embryogenesis. Additionally, we examined expression patterns in imaginal discs from third instar larvae, which will become the adult structures. Most isoforms are also expressed in the imaginal discs, with some showing unique transcript localization and spatial regulatory control. Thus, this report provides insight into the specific regions during Drosophila development that may require O-linked glycosylation in vivo as well as which isoforms may act cooperatively in certain tissues and which may be uniquely responsible for glycosylation in others. PMID- 16251382 TI - Predicting RNA folding thermodynamics with a reduced chain representation model. AB - Based on the virtual bond representation for the nucleotide backbone, we develop a reduced conformational model for RNA. We use the experimentally measured atomic coordinates to model the helices and use the self-avoiding walks in a diamond lattice to model the loop conformations. The atomic coordinates of the helices and the lattice representation for the loops are matched at the loop-helix junction, where steric viability is accounted for. Unlike the previous simplified lattice-based models, the present virtual bond model can account for the atomic details of realistic three-dimensional RNA structures. Based on the model, we develop a statistical mechanical theory for RNA folding energy landscapes and folding thermodynamics. Tests against experiments show that the theory can give much more improved predictions for the native structures, the thermal denaturation curves, and the equilibrium folding/unfolding pathways than the previous models. The application of the model to the P5abc region of Tetrahymena group I ribozyme reveals the misfolded intermediates as well as the native-like intermediates in the equilibrium folding process. Moreover, based on the free energy landscape analysis for each and every loop mutation, the model predicts five lethal mutations that can completely alter the free energy landscape and the folding stability of the molecule. PMID- 16251383 TI - Functional characterization of the SOFA delta ribozyme. AB - Molecular engineering has led to the development of a novel target-dependent riboswitch that increases deltaribozyme fidelity. This delta ribozyme possesses a specific on/off adapter (SOFA) that switches the cleavage activity from off (a "safety lock") to on solely in the presence of the desired RNA substrate. In this report, we investigate the influence of both the structure and the sequence of each domain of the SOFA module. Analysis of the cleavage activity, using a large collection of substrates and SOFA-ribozyme mutants, together with RNase H probing provided several insights into the nature of the sequence and the optimal design of each domain of the SOFA module. For example, we determined that (1) the optimal size of the blocker sequence, which keeps the ribozyme off in the absence of the substrate, is 4 nucleotides (nt); (2) a single nucleotide difference between the substrate and the biosensor domain, which is responsible for the initial binding of the substrate that subsequently switches the SOFA-ribozyme on, is sufficient to cause non-recognition of the appropriate substrate; (3) the stabilizer, which joins the 5' and 3' ends of the SOFA-ribozyme, plays only a structural role; and (4) the optimal spacer sequence, which serves to separate the binding regions of the biosensor and catalytic domain of the ribozyme on the substrate, is from 1 to 5 nt long. Together, these data should facilitate the design of more efficient SOFA-ribozymes with significant potential for many applications in gene-inactivation systems. PMID- 16251384 TI - Optimizing splinted ligation of highly structured small RNAs. AB - The synthesis of highly structured small RNAs containing nonstandard nucleotides is of high interest for structural and functional investigations. A general approach is the joining, by T4 DNA ligase-mediated splinted ligation, of two or more RNA fragments, each of which may contain its own set of modified nucleotides. The RNA fragments hybridize with a complementary DNA splint to form a ternary ligation-competent-complex (LCC), which is then turned over by the DNA ligase. We studied the formation of the LCC and its precursors using size exclusion chromatography combined with a fluorescence detector. The spatial proximity of two cyanine-dye-labeled RNA fragments in LCCs was detected by monitoring FRET. An observed correlation of LCC formation and ligation yields suggests the use of long splints to stabilize LCCs. Splint oligos of increasing length, which in general appear to reduce the number of different hybridization intermediate species found in a reaction mixture, were applied to the synthesis by T4-DNA-ligation of two highly structured target molecules, one a 73 mer tRNA, the other a 49 mer synthetic ribozyme. A stable LCC could be isolated and turned over with>95% ligation efficiency. In conclusion, the use of long splints presents a generally applicable means to overcome the low propensity of highly structured RNAs for hybridization, and thus to significantly improve ligation efficiencies. PMID- 16251386 TI - Further evidence that ribavirin interacts with eIF4E. AB - This commentary discusses the recent reports in RNA by Yan and colleagues and Westman and colleagues of the apparent failure of ribavirin to bind to recombinant eIF4E and inhibit 7-methyl guanosine cap-dependent exogenous mRNA translation of cell extracts in vitro. Measuring binding by using affinity chromatography of matrix-immobilized proteins and by using protein emission fluorescence spectroscopy in the presence of nucleotide ligands, as well as limitations of using cell extracts for the assessment of mechanisms of mRNA translation are discussed. Possible reasons for the discordant findings of Yan and colleagues and Westman and colleagues are suggested, and direct observation of the specific binding of ribavirin to eIF4E by using mass spectrometry is presented. PMID- 16251385 TI - Differences and similarities between Drosophila and mammalian 3' end processing of histone pre-mRNAs. AB - We used nuclear extracts from Drosophila Kc cells to characterize 3' end processing of Drosophila histone pre-mRNAs. Drosophila SLBP plays a critical role in recruiting the U 7 snRNP to the pre-mRNA and is essential for processing all five Drosophila histone pre-mRNAs. The Drosophila processing machinery strongly prefers cleavage after a fourth nucleotide following the stem-loop and favors an adenosine over pyrimidines in this position. Increasing the distance between the stem-loop and the HDE does not result in a corresponding shift of the cleavage site, suggesting that in Drosophila processing the U 7 snRNP does not function as a molecular ruler. Instead, SLBP directs the cleavage site close to the stem loop. The upstream cleavage product generated in Drosophila nuclear extracts contains a 3' OH, and the downstream cleavage product is degraded by a nuclease dependent on the U 7 snRNP, suggesting that the cleavage factor has been conserved between Drosophila and mammalian processing. A 2'O-methyl oligonucleotide complementary to the first 17 nt of the Drosophila U 7 snRNA was not able to deplete the U 7 snRNP from Drosophila nuclear extracts, suggesting that the 5' end of the Drosophila U 7 snRNA is inaccessible. This oligonucleotide selectively inhibited processing of only two Drosophila pre-mRNAs and had no effect on processing of the other three pre-mRNAs. Together, these studies demonstrate that although Drosophila and mammalian histone pre-mRNA processing share common features, there are also significant differences, likely reflecting divergence in the mechanism of 3' end processing between vertebrates and invertebrates. PMID- 16251387 TI - MAASE: an alternative splicing database designed for supporting splicing microarray applications. AB - Alternative splicing is a prominent feature of higher eukaryotes. Understanding of the function of mRNA isoforms and the regulation of alternative splicing is a major challenge in the post-genomic era. The development of mRNA isoform sensitive microarrays, which requires precise splice-junction sequence information, is a promising approach. Despite the availability of a large number of mRNAs and ESTs in various databases and the efforts made to align transcript sequences to genomic sequences, existing alternative splicing databases do not offer adequate information in an appropriate format to aid in splicing array design. Here we describe our effort in constructing the Manually Annotated Alternatively Spliced Events (MAASE) database system, which is specifically designed to support splicing microarray applications. MAASE comprises two components: (1) a manual/computational annotation tool for the efficient extraction of critical sequence and functional information for alternative splicing events and (2) a user-friendly database of annotated events that allows convenient export of information to aid in microarray design and data analysis. We provide a detailed introduction and a step-by-step user guide to the MAASE database system to facilitate future large-scale annotation efforts, integration with other alternative splicing databases, and splicing array fabrication. PMID- 16251389 TI - Immune-related conditions and immune-modulating medications as risk factors for non-Hodgkin's lymphoma: a case-control study. AB - In immunosuppressed or autoimmune disease states, disordered immune responses may lead to non-Hodgkin's lymphoma (NHL). In a US population-based case-control study of NHL (1998-2000), the authors collected personal histories of immune-related conditions and use of immune-modulating therapies as well as family histories of autoimmune conditions. The study included 1,321 NHL cases and 1,057 controls; only half received some questionnaire components. NHL was associated with Sjogren's syndrome (odds ratio (OR) = 13, 95% confidence interval (CI): 1.7, 100) and lupus (OR = 4.2, 95% CI: 1.2, 15). Two specific NHL subtypes were strongly associated with Sjogren's syndrome: salivary gland (OR = 290, 95% CI: 33, 2600) and marginal zone (OR = 75, 95% CI: 9.1, 610). NHL was less convincingly associated with receipt of an organ transplant (OR = 2.0, 95% CI: 0.4, 11). Other autoimmune conditions were too rare to evaluate or not associated with NHL. Corticosteroid use was unrelated to NHL (OR = 1.0, 95% CI: 0.8, 1.2), but methotrexate use was marginally associated (OR = 2.3, 95% CI: 0.7, 7.5). Family history of dermatomyositis was associated with NHL (7 cases vs. 0 controls, OR = infinite; two-sided p = 0.02), but dermatomyositis was absent in cases themselves. Family history of remaining conditions was unrelated to NHL. Results suggest that disordered immunity in some immune-related conditions can lead to NHL. PMID- 16251388 TI - Characteristics and regulatory elements defining constitutive splicing and different modes of alternative splicing in human and mouse. AB - Alternative splicing is a major contributor to genomic complexity, disease, and development. Previous studies have captured some of the characteristics that distinguish alternative splicing from constitutive splicing. However, most published work only focuses on skipped exons and/or a single species. Here we take advantage of the highly curated data in the MAASE database (see related paper in this issue) to analyze features that characterize different modes of splicing. Our analysis confirms previous observations about alternative splicing, including weaker splicing signals at alternative splice sites, higher sequence conservation surrounding orthologous alternative exons, shorter exon length, and more frequent reading frame maintenance in skipped exons. In addition, our study reveals potentially novel regulatory principles underlying distinct modes of alternative splicing and a role of a specific class of repeat elements (transposons) in the origin/evolution of alternative exons. These features suggest diverse regulatory mechanisms and evolutionary paths for different modes of alternative splicing. PMID- 16251390 TI - Model of risk of cortical cataract in the US population with exposure to increased ultraviolet radiation due to stratospheric ozone depletion. AB - The authors modeled the possible consequences for US cataract incidence of increases in ultraviolet B radiation due to ozone depletion. Data on the dose response relation between ocular exposure to ultraviolet B radiation and cortical cataract were derived from a population-based study (the Salisbury Eye Evaluation Project, Salisbury, Maryland) in which extensive data on cataract and ultraviolet radiation were collected in persons aged 65-84 years. Exposure estimates for the US population were derived using estimated ultraviolet radiation fluxes as a function of wavelength. US Census data were used to obtain the age, ethnicity, and sex distribution of the population. Predicted probabilities of cataract were derived from the age-, sex-, and ethnicity-specific ocular ultraviolet exposure data and were modeled under conditions of 5-20% ozone depletion. The analysis indicated that by 2050, the prevalence of cortical cataract will increase above expected levels by 1.3-6.9%. The authors estimate that with 5-20% ozone depletion, there will be 167,000-830,000 additional cases of cortical cataract by 2050. Because of the high prevalence of cataract in older persons, at a 2003 cost of 3,370 dollars per cataract operation, this increase could represent an excess cost of 563 million dollars to 2.8 billion dollars. PMID- 16251391 TI - Imaging of the patient with non-small cell lung cancer. AB - Lung cancer is the most common type of cancer and is the leading cause of cancer deaths in the United States for both men and women. Even though the 5-year survival rate of patients with lung cancer remains dismal at 14% for all cancer stages, treatments are improving and newer agents for lung cancer appear promising. Therefore, an accurate assessment of the extent of disease is critical to determine whether the patient is treated with surgical resection, radiation therapy, chemotherapy, or a combination of these modalities. Radiologic imaging plays an important role in the staging evaluation of the patient; however, radiologists need to be aware that there are also important differences in what each specialist needs from imaging to provide appropriate treatment. This article reviews the role of imaging in patients with non-small cell lung cancer, with an emphasis on the radiologic imaging findings relevant for each specialty. PMID- 16251392 TI - Interlobular septal thickening in idiopathic bronchiectasis: a thin-section CT study of 94 patients. AB - PURPOSE: To retrospectively establish the prevalence of interlobular septal thickening at thin-section computed tomography (CT) in patients with bronchiectasis and to retrospectively examine the relationship between septal thickening and various CT features of bronchiectasis and functional parameters. MATERIALS AND METHODS: Institutional review board approval or informed consent are not required for such a retrospective study. Thin-section CT scans of 94 patients (29 men, 65 women; age range, 18-82 years) with idiopathic bronchiectasis confirmed at thin-section CT and without any history or condition that causes bronchiectasis were assessed independently by two observers. The presence and signs of bronchiectasis, extent and distribution of septal thickening at global and lobar levels, and presence of emphysema were recorded. Univariate and multiple regression analyses were used to identify relationships between individual parameters of bronchiectasis, the extent of septal thickening, and functional indices. RESULTS: Interlobular septal thickening was present in 56 (60%) of 94 patients with idiopathic bronchiectasis, excluding those with trivial septal thickening (34 of 94, 36%). At a lobar level, 69% (514 of 748) of lobes with bronchiectasis had septal thickening. There was strong correlation between the extent of septal thickening and the extent (r = 0.61, P < .001) and severity (r = 0.54, P < .001) of bronchiectasis. Stepwise regression analysis showed a relationship between the extent of septal thickening and the extent of bronchiectasis (P < .001). Septal thickening was not linked to functional indices of obstruction or restriction. CONCLUSION: At thin-section CT, interlobular septal thickening is a frequent finding in patients with idiopathic bronchiectasis. PMID- 16251393 TI - Intraatrial repair of transposition of the great arteries: use of MR imaging after exercise to evaluate regional systemic right ventricular function. AB - PURPOSE: To prospectively assess regional systemic right ventricular (RV) function at rest and in response to exercise by using magnetic resonance (MR) imaging in patients who have undergone surgical correction at the atrial level for transposition of the great arteries (TGA). MATERIALS AND METHODS: Informed consent was obtained, and the medical review board approved this study. In 25 adult patients (mean age, 25.8 years +/- 4.7 [standard deviation]; 13 men) who had undergone correction for TGA (23.4 years +/- 4.9 after surgery) and 11 healthy volunteers (mean age, 27.4 years +/- 2.7; six men), systemic ventricular function was assessed with MR imaging (turbo field echo-planar imaging) at rest and during supine bicycle exercise. Regional wall thickness and wall thickening of the systemic RV were assessed and compared with those of the left ventricle in healthy volunteers by two investigators working together. Regional wall parameters were calculated by using the three-dimensional centerline method. Independent-samples t test and paired-samples t test were used for statistical analysis. RESULTS: Ejection fraction of the systemic RV did not increase after exercise (56% +/- 8 at rest to 55% +/- 7 after exercise, P = .196). Mean RV wall thickening was impaired in patients with TGA at all levels both at rest and in response to exercise (P < .05). Moreover, the free wall and the anterior wall of the systemic RV had a smaller end-systolic thickness and a diminished thickening at rest and after exercise compared with findings in the normal left ventricle (P < .05). CONCLUSION: The systemic RV of patients after intraatrial correction for TGA reveals regional functional disturbances at rest and in response to exercise. PMID- 16251394 TI - T staging of gastric cancer: role of multi-detector row CT. AB - PURPOSE: To evaluate retrospectively the accuracy of multi-detector row computed tomography (CT) in the assessment of serosal invasion in patients with gastric cancer. MATERIALS AND METHODS: The Ethics Committee does not require approval or informed consent for retrospective studies. Forty-one consecutive patients (24 men, 17 women; mean age, 68 years) with gastric cancer were included in this study. All patients were given 600 mL of tap water to drink and were positioned prone or supine on the scanning table. The detector row configuration included four detector rows, a section thickness of 1.25 mm, a pitch of 6, and a reconstruction interval of 0.63 mm. Transverse and multiplanar reconstruction images were simultaneously evaluated by two independent observers to assess the depth of tumor invasion in the gastric wall (ie, T stage). T staging at multi detector row CT was compared with T staging at histologic evaluation (reference standard), which was performed by means of surgical or histologic examination of the resected specimen. We also calculated the sensitivity, specificity, and accuracy of multi-detector row CT for each observer in the assessment of serosal invasion. RESULTS: Analysis of interobserver agreement showed substantial or almost perfect agreement (nonweighted kappa value of 0.78 and weighted kappa value of 0.85). Correct assessment of gastric wall invasion was 80% and 85% for observers 1 and 2, respectively. The sensitivity, specificity, and accuracy of multi-detector row CT in the assessment of serosal invasion were 90%, 95%, and 93%, respectively, for observer 1 and 80%, 97%, and 93%, respectively, for observer 2. Overstaging occurred in six patients, and understaging occurred in five patients. All understaged tumors were scirrhous subtype gastric cancer. CONCLUSION: Multi-detector row CT scanning of patients with gastric cancer gave 93% accuracy in the assessment of serosal invasion in patients with gastric cancer. PMID- 16251395 TI - Clinical course of knees with asymptomatic meniscal abnormalities: findings at 2 year follow-up after MR imaging-based diagnosis. AB - PURPOSE: To prospectively evaluate the clinical course of asymptomatic meniscal lesions diagnosed by using magnetic resonance (MR) imaging. MATERIALS AND METHODS: Institutional review board approval and informed consent were obtained. The clinical courses of meniscal lesions in 84 asymptomatic knees (in 48 men and 36 women; mean age, 43.6 years; age range, 18-73 years) were assessed. Thirty-one asymptomatic meniscal lesions were depicted among the 84 knees at MR imaging. The follow-up period was at least 2 years (mean, 29.8 months; range, 24-36 months). Knee pain, stiffness, and function during daily and sports activities were assessed by using a visual analogue scale (VAS), on which a score of 0 indicated no pain or complaints and a score of 100 indicated maximal pain and/or complaints. The chi2 test was used for statistical analysis. RESULTS: At follow up, 12 (39%) of the 31 patients with and 10 (19%) of the 53 patients without meniscal lesions reported having knee pain (P = .046). Nine (29%) patients with and five (9%) patients without meniscal lesions reported having knee stiffness (P = .02). Ten (32%) patients with and five (9%) patients without meniscal lesions reported having impaired function during daily activities (P = .008). Thirteen (42%) patients with and 15 (28%) patients without meniscal lesions reported having impaired function during sports activities (P = .20). Mean VAS scores ranged from 22 to 30; mean VAS scores for pain, stiffness, impaired daily function, and impaired sports function were 26, 22, 27, and 30, respectively. None of the initially asymptomatic knees was treated with surgery during the follow-up period. CONCLUSION: The clinical course of knees with initially asymptomatic meniscal lesions, compared with the clinical course of knees without meniscal lesions, is characterized by an increased frequency of complaints. However, severities of pain and knee function impairment remain low. PMID- 16251396 TI - Prostate cancer: three-dimensional sonoelastography for in vitro detection. AB - PURPOSE: To prospectively evaluate the accuracy of three-dimensional (3D) sonoelastographic imaging, relative to that of gray-scale ultrasonography (US), in the in vitro detection of prostate cancer. MATERIALS AND METHODS: The study was approved by the institutional review board and was HIPAA compliant. Informed consent was obtained from all patients. Nineteen prostatectomy specimens from patients aged 46-70 years with biopsy-proved prostate cancer were scanned in three dimensions by using conventional B-mode US and sonoelastography with vibrations of more than 100 Hz. Step-sectioned whole-mount histologic specimens were used to create a 3D volume of the prostate and the tumors within it. B-mode US scans and regions of low vibration on the sonoelastographic images (hard regions) were formatted in three dimensions. The lesions in the 19 cases were classified into two groups, as follows: G1 lesions were pathologically confirmed tumors with a volume of at least 1.0 cm3, and G2 lesions were pathologically confirmed tumors smaller than 1.0 cm3. G1 lesions were evaluated with B-mode US and sonoelastography and classified as true-positive, false-positive, true negative, or false-negative; G2 lesions were evaluated only with sonoelastography. Findings at histologic examination were used as the reference standard. True-positive findings necessitated 3D lesion correlation between pathologic and imaging data. Conventional definitions of accuracy and sensitivity were used for statistical analysis. RESULTS: For G1 lesions (seven lesions with a volume of at least 1.0 cm3), sonoelastography had an accuracy of 55% and a sensitivity of 71% and B-mode US had an accuracy of 17% and a sensitivity of 29%. The mean tumor volume was 3.1 cm3 +/- 2.1 (standard deviation). For G2 lesions (22 lesions with a volume of less than 1.0 cm3), the mean tumor volume was 0.32 cm3 +/- 0.21. Sonoelastography had an accuracy of 34% and a sensitivity of 41%; there were six false-positive findings. CONCLUSION: Sonoelastography performed considerably better than did gray-scale US in the depiction of prostate cancer for tumors with volumes of more than 1 cm3. PMID- 16251397 TI - Arterial blood supply to the posterior aspect of segment IV of the liver from the caudate branch: demonstration at CT after iodized oil injection. AB - PURPOSE: To retrospectively evaluate the arterial blood supply to the posterior aspect of segment IV of the liver with computed tomography (CT) after transcatheter arterial chemoembolization (TACE) with iodized oil through the caudate arterial branch of the liver for treatment of hepatocellular carcinoma (HCC). MATERIALS AND METHODS: Institutional review board approval and patient informed consent were not required for this retrospective study. Twenty-four patients (11 men and 13 women; mean age, 68 years) with HCC originating in the caudate lobe (n = 23) or posterior aspect of segment IV (n = 1) were selected. TACE of the caudate arterial branch was performed in all patients, including one patient with HCC in the posterior aspect of segment IV who underwent TACE of the caudate arterial branch after CT helped confirm that iodized oil was not distributed in the tumor after TACE of the medial segmental artery. The distribution of iodized oil in the posterior aspect of segment IV was analyzed with CT 1 week after TACE. The number and origin of all arteries supplying the caudate lobe and the number of arteries embolized were determined. RESULTS: Thirty-three caudate arterial branches were embolized. Twenty-nine branches were derived from the right hepatic artery and four were derived from the left hepatic artery. A single branch was seen in 17 patients, two branches were seen in five, and three branches were seen in two. Eight patients simultaneously underwent additional TACE of branches of the right hepatic artery (n = 6) or right inferior phrenic artery (n = 2). At CT, iodized oil was seen to be distributed entirely (n = 19) or partially (n = 5) in the caudate lobe. Distribution of iodized oil at the posterior aspect of segment IV was observed in 16 patients (67%), including 13 (54%) whose caudate arterial branches were derived entirely from the right hepatic artery. CONCLUSION: The results of this study suggest that the caudate arterial branch, which is mainly derived from the right hepatic artery, frequently supplies the posterior aspect of segment IV. This knowledge is important for managing HCC in the posterior aspect of segment IV by means of TACE. PMID- 16251398 TI - Osteoarthritis of the knee: comparison of MR imaging findings with radiographic severity measurements and pain in middle-aged women. AB - PURPOSE: To prospectively compare magnetic resonance (MR) imaging-defined abnormalities of osteoarthritis (OA) of the knee with radiographic severity measurements of OA of the knee and self-reported pain. MATERIALS AND METHODS: This study was approved by the institutional review board of University of Michigan. Informed consent was obtained for this HIPAA-compliant study. Knee MR imaging was performed in 117 women (mean age, 46 years; range, 32-56 years) from a community-based arthritis study (n = 1053) with 30 women in each of four categories: (a) no pain and no OA of the knee, (b) no pain and OA of the knee, (c) pain and no OA of the knee, and (d) pain and OA of the knee. OA of the knee was defined from radiographs. Two hundred thirty-two eligible knees had Kellgren Lawrence scores for OA of the knee as follows: grade 0, 115 (49.6%); grade 1, 33 (14.2%); grade 2, 66 (28.4%); grade 3, 17 (7.3%); and grade 4, one (0.4%). MR images were assessed for location and severity of defects of cartilage, bone marrow edema (BME), osteophytes, subchondral cysts, sclerosis, meniscal and/or ligamentous tears, joint effusion, synovial cysts, and synovitis. MR imaging findings were compared with radiographic severity of OA of the knee (Kellgren Lawrence scale) and self-reported pain with analysis of variance, t tests, and contingency table analyses. RESULTS: Defects of cartilage (higher than grade IIA) were found in 75% of knees; BME was found in 57% of knees (<1 cm, 41%; >1 cm, 16%). Large BME lesions were common in the pain and OA of the knee group (P = .001); this group was significantly more likely to have defects of cartilage (P = .001); meniscal tears (P = .001); and osteophytes, subchondral cysts, sclerosis, joint effusion, and synovitis (P < .001). Defects of cartilage, osteophytes, sclerosis, meniscal or ligamentous tears, joint effusion, and synovitis were strongly related to increasing Kellgren-Lawrence grade (P < .001). CONCLUSION: In middle-aged women, there were significant associations between pain, radiographic severity of OA of the knee, and seven MR imaging-identified parameters. PMID- 16251399 TI - Blunt cerebrovascular injury in patients with blunt multiple trauma: diagnostic accuracy of duplex Doppler US and early CT angiography. AB - PURPOSE: To retrospectively evaluate the frequency of blunt cerebrovascular injury (BCVI) in patients with multiple trauma and to retrospectively compare the diagnostic accuracy of duplex Doppler ultrasonography (US) and computed tomographic (CT) angiography by using clinical follow-up and subsequent imaging as reference standards. MATERIALS AND METHODS: The institutional review board approved this study; informed consent was not required. Charts and images of consecutive patients treated for multiple trauma (injury severity score, >16) between January 1998 and October 2003 were reread by an experienced radiologist. Until October 2002, subjects were screened for BCVI with US. Since November 2002, patients underwent CT angiography of the carotid and vertebral arteries. Sensitivity and specificity of US and CT angiography were calculated with 95% confidence intervals (CIs). RESULTS: The early cohort included 1471 patients (mean age, 35.8 years +/- 17.7 [standard deviation]), and the late cohort included 407 patients (mean age, 39.2 years +/- 18.8). US depicted five blunt vessel injuries but later missed another eight, which led to cerebral ischemia. With a BCVI frequency of 0.9%, sensitivity and specificity of US were 38.5% (95% CI: 13.9%, 68.4%) and 100% (lower 95% confidence limit, 99.7%), respectively. In the second cohort, the BCVI rate was 2.7%. CT angiography depicted BCVI in 11 patients, with a sensitivity of 100% (lower 95% confidence limit, 71.5%), but produced one false-positive result. CONCLUSION: Injuries to the cervical arteries among blunt trauma patients are more common than previously reported. Duplex Doppler US has inadequate sensitivity to help rule out this condition. The notable morbidity with missed dissections warrants routine contrast material enhanced studies of the carotid and vertebral vessels if patients are scheduled for CT of the cervical spine. PMID- 16251400 TI - In vivo and in vitro studies of Mgs1 suggest a link between genome instability and Okazaki fragment processing. AB - The non-essential MGS1 gene of Saccharomyces cerevisiae is highly conserved in eukaryotes and encodes an enzyme containing both DNA-dependent ATPase and DNA annealing activities. MGS1 appears to function in post-replicational repair processes that contribute to genome stability. In this study, we identified MGS1 as a multicopy suppressor of the temperature-sensitive dna2Delta405N mutation, a DNA2 allele lacking the N-terminal 405 amino acid residues. Mgs1 stimulates the structure-specific nuclease activity of Rad27 (yeast Fen1 or yFen1) in an ATP dependent manner. ATP binding but not hydrolysis was sufficient for the stimulatory effect of Mgs1, since non-hydrolyzable ATP analogs are as effective as ATP. Suppression of the temperature-sensitive growth defect of dna2Delta405N required the presence of a functional copy of RAD27, indicating that Mgs1 suppressed the dna2Delta405N mutation by increasing the activity of yFen1 (Rad27) in vivo. Our results provide in vivo and in vitro evidence that Mgs1 is involved in Okazaki fragment processing by modulating Fen1 activity. The data presented raise the possibility that the absence of MGS1 may impair the processing of Okazaki fragments, leading to genomic instability. PMID- 16251402 TI - Covariate-adjusted varying coefficient models. AB - Covariate-adjusted regression was recently proposed for situations where both predictors and response in a regression model are not directly observed, but are observed after being contaminated by unknown functions of a common observable covariate. The method has been appealing because of its flexibility in targeting the regression coefficients under different forms of distortion. We extend this methodology proposed for regression into the framework of varying coefficient models, where the goal is to target the covariate-adjusted relationship between longitudinal variables. The proposed method of covariate-adjusted varying coefficient model (CAVCM) is illustrated with an analysis of a longitudinal data set containing calcium absorbtion and intake measurements on 188 subjects. We estimate the age-dependent relationship between these two variables adjusted for the covariate body surface area. Simulation studies demonstrate the flexibility of CAVCM in handling different forms of distortion in the longitudinal setting. PMID- 16251403 TI - Rous-Whipple Award Lecture. The Alzheimer's brain: finding out what's broken tells us how to fix it. PMID- 16251401 TI - Zinc finger nucleases: custom-designed molecular scissors for genome engineering of plant and mammalian cells. AB - Custom-designed zinc finger nucleases (ZFNs), proteins designed to cut at specific DNA sequences, are becoming powerful tools in gene targeting--the process of replacing a gene within a genome by homologous recombination (HR). ZFNs that combine the non-specific cleavage domain (N) of FokI endonuclease with zinc finger proteins (ZFPs) offer a general way to deliver a site-specific double strand break (DSB) to the genome. The development of ZFN-mediated gene targeting provides molecular biologists with the ability to site-specifically and permanently modify plant and mammalian genomes including the human genome via homology-directed repair of a targeted genomic DSB. The creation of designer ZFNs that cleave DNA at a pre-determined site depends on the reliable creation of ZFPs that can specifically recognize the chosen target site within a genome. The (Cys2His2) ZFPs offer the best framework for developing custom ZFN molecules with new sequence-specificities. Here, we explore the different approaches for generating the desired custom ZFNs with high sequence-specificity and affinity. We also discuss the potential of ZFN-mediated gene targeting for 'directed mutagenesis' and targeted 'gene editing' of the plant and mammalian genome as well as the potential of ZFN-based strategies as a form of gene therapy for human therapeutics in the future. PMID- 16251404 TI - Opiate abuse and viral replication in hepatitis C. PMID- 16251405 TI - Origin of interstitial fibroblasts in an accelerated model of angiotensin II induced renal fibrosis. AB - To determine whether previous renal injury accelerates the progression of glomerulosclerosis and interstitial fibrosis, we examined the effect of treating rats with angiotensin II after Habu venom injury. After initiating disease, we examined the origin of interstitial myofibroblasts by locating alpha-smooth muscle actin (alpha-SMA)-positive and Na+,K+-ATPase-positive cells relative to interstitial space, tubular epithelial cells, the tubular basement membrane (TBM), and vascular structures. Tubular epithelial-mesenchymal transition was also assessed by examining TBM integrity and by using Texas Red (TR)-dextran in intravital tracking experiments. The staining of alpha-SMA-positive myofibroblasts dramatically increased in peritubular interstitial spaces 48 hours after Habu venom plus angiotensin II, particularly in and around perivascular and periglomerular regions, while tubular epithelial cells were alpha-SMA-negative. Na+,K+-ATPase-positive and TR-dextran-labeled cells were restricted to the tubular epithelium and excluded from the interstitium. By 7 and 14 days, expanded interstitial space contained only alpha-SMA-positive myofibroblasts without TR dextran endocytic particles. Epithelium of atrophic tubules containing TR-dextran remained confined by surrounding interstitium and myofibroblasts. These studies indicate that early expansion of alpha-SMA-positive cells in the interstitium and loss of tubular area occur via encroachment of interstitial myofibroblasts from perivascular into atrophic tubular spaces rather than via epithelial-mesenchymal transition and migration of tubular cells through the TBM into the interstitium. PMID- 16251406 TI - Conditional ablation of macrophages halts progression of crescentic glomerulonephritis. AB - The presence of macrophages in inflamed glomeruli of rat kidney correlates with proliferation and apoptosis of resident glomerular mesangial cells. We assessed the contribution of inflammatory macrophages to progressive renal injury in murine crescentic glomerulonephritis (GN). Using a novel transgenic mouse (CD11b DTR) in which tissue macrophages can be specifically and selectively ablated by minute injections of diphtheria toxin, we depleted renal inflammatory macrophages through days 15 and 20 of progressive crescentic GN. Macrophage depletion reduced the number of glomerular crescents, improved renal function, and reduced proteinuria. Morphometric analysis of renal tubules and interstitium revealed a marked attenuation of tubular injury that was associated with reduced proliferation and apoptosis of tubular cells. The population of interstitial myofibroblasts decreased after macrophage depletion and interstitial fibrosis also decreased. In the presence of macrophages, interstitial myofibroblasts exhibited increased levels of both proliferation and apoptosis, suggesting that macrophages act to support a population of renal myofibroblasts in a high turnover state and in matrix deposition. Finally, deletion of macrophages reduced CD4 T cells in the diseased kidney. This study demonstrates that macrophages are key effectors of disease progression in crescentic GN, acting to regulate parenchymal cell populations by modulating both cell proliferation and apoptosis. PMID- 16251407 TI - Susceptibility of signal transducer and activator of transcription-1-deficient mice to pulmonary fibrogenesis. AB - The signal transducer and activator of transcription (Stat)-1 mediates growth arrest and apoptosis. We postulated that lung fibrosis characterized by excessive proliferation of lung fibroblasts would be enhanced in Stat1-deficient (Stat1-/-) mice. Two weeks after bleomycin aspiration (3 U/kg), Stat1-/- mice exhibited a more severe fibroproliferative response and significantly elevated total lung collagen compared to wild-type mice. Growth factors [epidermal growth factor (EGF) or platelet-derived growth factor (PDGF)] enhanced [3H]thymidine uptake in lung fibroblasts isolated from Stat1-/- mice compared to wild-type mice. Interferon (IFN)-gamma, which signals growth arrest via Stat1, inhibited EGF- or PDGF-stimulated mitogenesis in wild-type fibroblasts but enhanced [3H]thymidine uptake in Stat1-/- fibroblasts. Moreover, IFN-gamma treatment in the absence of growth factors induced a concentration-dependent increase in [3H]thymidine uptake in Stat1-/- but not wild-type fibroblasts. Mitogen-activated protein kinase (ERK 1/2) phosphorylation in response to PDGF or EGF did not differ among Stat1-/- and wild-type fibroblasts. However, Stat3 phosphorylation induced by PDGF, EGF, or IFN-gamma increased twofold in Stat1-/- fibroblasts compared to wild-type fibroblasts. Our findings indicate that Stat1-/- mice are more susceptible to bleomycin-induced lung fibrosis than wild-type mice due to 1) enhanced fibroblast proliferation in response to growth factors (EGF and PDGF), 2) stimulation of fibroblast growth by a Stat1-independent IFN-gamma signaling pathway, and 3) increased activation of Stat3. PMID- 16251408 TI - Natural killer T (NKT) cells attenuate bleomycin-induced pulmonary fibrosis by producing interferon-gamma. AB - Pulmonary fibrosis is a progressive illness characterized by interstitial fibrosis. Although the precise mechanism for pulmonary fibrosis is not completely understood, an immune response involving interferon (IFN)-gamma appears to play a role. Therefore, we examined the functional roles of natural killer T (NKT) cells, which produce IFN-gamma and interleukin-4 on activation, in bleomycin induced pulmonary fibrosis. In NKT cell-deficient mice, pulmonary fibrosis was worse in terms of histology, hydroxyproline levels, and mortality than in control mice. The transforming growth factor (TGF)-beta1 levels were higher in the lung after injecting bleomycin, and blockade of TGF-beta1 by neutralizing monoclonal antibody attenuated the pulmonary fibrosis in CD1d-/- mice. In contrast, the production of IFN-gamma was reduced in lungs from CD1d-/- mice. Moreover, the adoptive transfer of NKT cells into CD1d-/- mice increased IFN-gamma and reduced TGF-beta1 production, attenuating pulmonary fibrosis. An in vitro assay demonstrated that IFN-gamma was involved in suppressing TGF-beta1 production in cells collected from bronchoalveolar lavage. The adoptive transfer of NKT cells from IFN-gamma-/- mice did not reverse pulmonary fibrosis or TGF-beta1 production in lungs of CD1d-/- mice whereas NKT cells from B6 control mice attenuated fibrosis and reduced TGF-beta1 production. In conclusion, IFN-gamma-producing NKT cells play a novel anti-fibrotic role in pulmonary fibrosis by regulating TGF beta1 production. PMID- 16251409 TI - Gene expression profiles reveal increased mClca3 (Gob5) expression and mucin production in a murine model of asbestos-induced fibrogenesis. AB - To elucidate genes important in development or repair of asbestos-induced lung diseases, gene expression was examined in mice after inhalation of chrysotile asbestos for 3, 9, and 40 days. We identified changes in the expression of genes linked to proliferation (cyclin B2, CDC20, and CDC28 protein kinase regulatory subunit 2), inflammation (CCL9, CCL6, complement component 1, chitinase3-like 3, TNF superfamily member 10, and IL-1B), and matrix remodeling (MMP12, MMP3, integrin alphaX, and cathepsins K, Z, B, and S). The most highly induced gene at all time points was mclca3 (gob5), a putative calcium-activated chloride channel involved in the regulation of mucus production and/or secretion. Using histochemistry, we demonstrated accumulation of mucus and increased mClca3 protein in the bronchiolar epithelium of asbestos-exposed mice at all time points but peaking at 9 days. Cytokine levels (interleukin-1beta, interleukin-4, interleukin-6) in bronchoalveolar lavage fluid also increased at 9 days, suggesting Th2-mediated immunity may play a role in asbestos-induced mucus production. In contrast, levels of cathepsin K, a potent elastase, increased between 3 and 40 days at both the mRNA and protein levels, localizing primarily in CD45-positive leukocytes and interstitial cells. Identification of genes involved in lung injury and remodeling after asbestos exposure could aid in defining mechanisms of airborne particulate-induced disease and in developing therapeutic strategies. PMID- 16251410 TI - Novel function for vascular endothelial growth factor receptor-1 on epidermal keratinocytes. AB - Vascular endothelial growth factor (VEGF-A), a potent stimulus for angiogenesis, is up-regulated in the skin after wounding. Although studies have shown that VEGF is important for wound repair, it is unclear whether this is based solely on its ability to promote angiogenesis or if VEGF can also promote healing by acting directly on non-endothelial cell types. By immunohistochemistry and reverse transcriptase-polymerase chain reaction, expression of VEGF receptor-1 (VEGFR-1), but not VEGFR-2, was detected in murine keratinocytes during wound repair and in normal human epidermal keratinocytes (NHEKs). The presence of VEGF receptors on NHEKs was verified by binding studies with 125I-VEGF. In vitro, VEGF stimulated the proliferation of NHEKs, an effect that could be blocked by treatment with neutralizing VEGFR-1 antibodies. A role for VEGFR-1 in keratinocytes was also shown in vivo because treatment of excisional wounds with neutralizing VEGFR-1 antibodies delayed re-epithelialization. Treatment with anti-VEGFR-1 antibodies also reduced the number of proliferating keratinocytes at the leading edge of the wound, suggesting that VEGF sends a proliferative signal to these cells. Together, these data describe a novel role for VEGFR-1 in keratinocytes and suggest that VEGF may play several roles in cutaneous wound repair. PMID- 16251411 TI - Increased and prolonged pulmonary fibrosis in surfactant protein C-deficient mice following intratracheal bleomycin. AB - Recent reports have linked mutations in the surfactant protein C gene (SFTPC) to familial forms of pulmonary fibrosis, but it is uncertain whether deficiency of mature SP-C contributes to disease pathogenesis. In this study, we evaluated bleomycin-induced lung fibrosis in mice with genetic deletion of SFTPC. Compared with wild-type (SFTPC+/+) controls, mice lacking surfactant protein C (SFTPC-/-) had greater lung neutrophil influx at 1 week after intratracheal bleomycin, greater weight loss during the first 2 weeks, and increased mortality. At 3 and 6 weeks after bleomycin, lungs from SFTPC-/- mice had increased fibroblast numbers, augmented collagen accumulation, and greater parenchymal distortion. Furthermore, resolution of fibrosis was delayed. Although remodeling was near complete in SFTPC+/+ mice by 6 weeks, SFTPC-/- mice did not return to baseline until 9 weeks after bleomycin. By terminal dUTP nick-end labeling staining, widespread cell injury was observed in SFTPC-/- and SFTPC+/+ mice 1 week after bleomycin; however, ongoing apoptosis of epithelial and interstitial cells occurred in lungs of SFTPC-/- mice, but not SFTPC+/+ mice, 6 weeks after bleomycin. Thus, SP-C functions to limit lung inflammation, inhibit collagen accumulation, and restore normal lung structure after bleomycin. PMID- 16251412 TI - Mouse fetal liver cells in artificial capillary beds in three-dimensional four compartment bioreactors. AB - Bioreactors containing porcine or adult human hepatocytes have been used to sustain acute liver failure patients until liver transplantation. However, prolonged function of adult hepatocytes has not been achieved due to compromised proliferation and viability of adult cells in vitro. We investigated the use of fetal hepatocytes as an alternative cell source in bioreactors. Mouse fetal liver cells from gestational day 17 possessed intermediate differentiation and function based on their molecular profile. When cultured in a three-dimensional four compartment hollow fiber-based bioreactor for 3 to 5 weeks these cells formed neo tissues that were characterized comprehensively. Albumin liberation, testosterone metabolism, and P450 induction were demonstrated. Histology showed predominant ribbon-like three-dimensional structures composed of hepatocytes between hollow fibers. High positivity for proliferating cell nuclear antigen and Ki-67 and low positivity for terminal dUTP nick-end labeling indicated robust cell proliferation and survival. Most cells within these ribbon arrangements were albumin-positive. In addition, cells in peripheral zones were simultaneously positive for alpha-fetoprotein, cytokeratin-19, and c-kit, indicating their progenitor phenotype. Mesenchymal components including endothelial, stellate, and smooth muscle cells were also observed. Thus, fetal liver cells can survive, proliferate, differentiate, and function in a three-dimensional perfusion culture system while maintaining a progenitor pool, reflecting an important advance in hepatic tissue engineering. PMID- 16251413 TI - Predisposition to colorectal cancer in rats with resolved colitis: role of cyclooxygenase-2-derived prostaglandin d2. AB - Colitis markedly increases the risk of developing colon cancer, but the underlying mechanisms are not fully understood. In a rat model of colitis, alterations in epithelial secretion, proliferation, and barrier function persist long after healing has occurred. In the present study, we examined whether rats that have recovered from a bout of colitis are more susceptible to preneoplastic lesions and whether this susceptibility is mediated by cyclooxygenase (COX)-2 derived prostaglandin (PG) D2. Colitis was induced by intracolonic administration of trinitrobenzenesulfonic acid. Six weeks later, weekly treatment with the carcinogen azoxymethane was initiated. Postcolitis rats exhibited significantly more aberrant crypt foci after azoxymethane treatment than controls. The postcolitis rats also exhibited markedly increased colonic PGD2 synthesis and elevated COX-2, H-PGD synthase, and beta-catenin expression. Treatment for 1 week with a selective COX-2 inhibitor or with a selective PGD2 receptor (DP1) antagonist significantly reduced susceptibility of postcolitis rats to aberrant crypt foci development, beta-catenin expression, and mucosal thickness. The results from this animal model suggest that prolonged elevation of COX-2-derived PGD2 synthesis after resolution of colitis may contribute significantly to colitis-associated increases in colon cancer incidence. PGD2 may therefore represent a rational target for therapies directed at reducing the incidence of colitis-associated colorectal cancer. PMID- 16251414 TI - Matched skin and sentinel lymph node samples of melanoma patients reveal exclusive migration of mature dendritic cells. AB - Mature and immature myeloid dendritic cells (DCs) are thought to differentially modulate T-cell responses in secondary lymphoid tissues. Although mature DCs are believed to induce T-cell activation under proinflammatory conditions, immature DCs are believed to maintain a state of T-cell tolerance under steady state conditions. However, little is known about the actual activation state of human DCs under these different conditions. Here, we compare the frequency and activation state of human DCs between matched skin and sentinel lymph node (SLN) samples, after intradermal administration of either granulocyte/macrophage colony stimulating factor (GM-CSF) or saline, at the excision site of stage I primary melanoma. Although DCs remained immature (CD1a+CD83-) and mostly situated in the epidermis of the saline-injected skin (fully consistent with a quiescent steady state), mature (CD1a+CD83+) DC frequencies significantly increased in the GM-CSF injected skin and correlated with the number of mature DCs in the SLN, indicative of increased DC migration. Interestingly, irrespective of GM-CSF or saline administration, all CD1a+ myeloid DCs in the SLN were phenotypically mature (ie, CD83+). These data are indicative of migration of small numbers of phenotypically mature DCs to lymph nodes under steady state conditions. PMID- 16251415 TI - Bacillus anthracis edema toxin causes extensive tissue lesions and rapid lethality in mice. AB - Bacillus anthracis edema toxin (ET), an adenylyl cyclase, is an important virulence factor that contributes to anthrax disease. The role of ET in anthrax pathogenesis is, however, poorly understood. Previous studies using crude toxin preparations associated ET with subcutaneous edema, and ET-deficient strains of B. anthracis showed a reduction in virulence. We report the first comprehensive study of ET-induced pathology in an animal model. Highly purified ET caused death in BALB/cJ mice at lower doses and more rapidly than previously seen with the other major B. anthracis virulence factor, lethal toxin. Observations of gross pathology showed intestinal intralumenal fluid accumulation followed by focal hemorrhaging of the ileum and adrenal glands. Histopathological analyses of timed tissue harvests revealed lesions in several tissues including adrenal glands, lymphoid organs, bone, bone marrow, gastrointestinal mucosa, heart, and kidneys. Concomitant blood chemistry analyses supported the induction of tissue damage. Several cytokines increased after ET administration, including granulocyte colony stimulating factor, eotaxin, keratinocyte-derived cytokine, MCP-1/JE, interleukin 6, interleukin-10, and interleukin-1beta. Physiological measurements also revealed a concurrent hypotension and bradycardia. These studies detail the extensive pathological lesions caused by ET and suggest that it causes death due to multiorgan failure. PMID- 16251416 TI - Ex vivo expanded dendritic cells home to T-cell zones of lymphoid organs and survive in vivo after allogeneic bone marrow transplantation. AB - Little is known about adoptive transfer of allogeneic ex vivo expanded dendritic cells (eDCs). We investigated the trafficking pattern of eDCs in mice after allogeneic bone marrow transplantation by using bioluminescence imaging. eDCs were expanded from bone marrow precursors in the presence of GM-CSF, interleukin 4, and Flt3L and retrovirally transduced to express luciferase (luc) and green fluorescence protein (gfp). Flow cytometry showed polyclonal DC populations after expansion that consisted of CD11c+CD11b+ and CD11c-CD11b+ cells that co-expressed CD40, CD80, CD86, and MHCII. eDCs were functional in mixed lymphocyte reactions and produced tumor necrosis factor-alpha on phytohemagglutinin stimulation. The eDCs were then injected intravenously into BALB/c recipient mice that had received allogeneic bone marrow transplantation 6 weeks previously. On day 1 after transfer, eDCs were detected by bioluminescence imaging throughout the lungs and spleen. In the later course, signals were observed throughout thymus, lower abdomen, and spleen throughout a period of more than 42 days. Immunofluorescence microscopy confirmed CD11c positivity on the gfp+ donor cells, which localized in T-cell zones of mesenteric lymph nodes, Peyer's patches, spleen, and thymus. These findings are important for adoptive immunotherapies because they indicate that eDCs migrate efficiently in vivo and are capable of surviving long term. PMID- 16251417 TI - Morphine withdrawal enhances hepatitis C virus replicon expression. AB - We previously demonstrated that morphine enhances hepatitis C virus (HCV) replication in human hepatic cells. Here we describe the impact of morphine withdrawal (MW), a recurrent event during the course of opioid abuse, on HCV replicon expression in human hepatic cells. MW enhanced both viral RNA and protein expression in HCV replicon cells. Blocking opioid receptors by treatment with naloxone after morphine cessation (precipitated withdrawal, PW) induced greater HCV replicon expression than MW. Investigation of the mechanism responsible for MW- or PW-mediated HCV enhancement showed that both MW and PW inhibited the expression of endogenous interferon-alpha (IFN-alpha) in the hepatic cells. This down-regulation of intracellular IFN-alpha expression was due to the negative impact of MW or PW on IFN-alpha promoter activation and on the expression of IFN regulatory factor 7 (IRF-7), a strong transactivator of the IFN alpha promoter. In addition, both MW and PW inhibited the anti-HCV ability of recombinant IFN-alpha in the hepatic cells. These in vitro observations support the concept that opioid abuse favors HCV persistence in hepatic cells by suppressing IFN-alpha-mediated intracellular innate immunity and contributes to the development of chronic HCV infection. PMID- 16251418 TI - Osteoclasts from patients with autosomal dominant osteopetrosis type I caused by a T253I mutation in low-density lipoprotein receptor-related protein 5 are normal in vitro, but have decreased resorption capacity in vivo. AB - Autosomal dominant osteopetrosis type I (ADOI) is presumably caused by gain-of function mutations in the LRP5 gene. Patients with a T253I mutation in LRP5 have a high bone mass phenotype, characterized by increased mineralizing surface index but abnormally low numbers of small osteoclasts. To investigate the effect of the T253I mutation in LRP5 on osteoclasts, we isolated CD14+ monocytes from ADOI patients and assessed their ability to generate osteoclasts when treated with RANKL and M-CSF compared to that of age- and sex-matched control osteoclasts. We found normal osteoclastogenesis, expression of osteoclast markers, morphology, and localization of proteins involved in bone resorption, such as ClC-7 and cathepsin K. The ability to resorb bone was also normal. In vivo, we compared the bone resorption and bone formation response to T3 in ADOI patients and age- and sex-matched controls. We found attenuated resorptive response to T3 stimulation, despite a normal bone formation response, in alignment with the reduced number of osteoclasts in vivo. These data demonstrate that ADOI osteoclasts are normal with respect to all aspects investigated in vitro. We speculate that the mutations causing ADOI alter the osteoblastic phenotype toward a smaller potential for supporting osteoclastogenesis. PMID- 16251419 TI - Temporal exposure of cryptic collagen epitopes within ischemic muscle during hindlimb reperfusion. AB - Chronic limb-threatening ischemia is a devastating disease with limited surgical options. However, inducing controlled angiogenesis and enhancing reperfusion holds therapeutic promise. To gain a better understanding of the mechanisms that contribute to limb reperfusion, we examined the temporal biochemical and structural changes occurring within the extracellular matrix of ischemic skeletal muscle. Both the latent and active forms of MMP-2 and -9 significantly increased during the active phase of limb reperfusion. Moreover, small but significant alterations in tissue inhibitors of metalloproteinase levels also occurred during a similar time course, consistent with a net increase in extracellular matrix remodeling. This temporal increase in MMP activity coincided with enhanced exposure of the unique HU177 cryptic collagen epitope. Although the HUIV26 cryptic collagen epitope has been implicated in angiogenesis, little is known concerning such epitopes within ischemic muscle tissue. Here, we provide the first evidence that a functionally distinct cryptic collagen epitope (HU177) is temporally exposed in ischemic muscle tissue during the active phase of reperfusion. Interestingly, the exposure of the HU177 epitope was greatly diminished in MMP-9 null mice, corresponding with significantly reduced limb reperfusion. Therefore, the regulated exposure of a unique cryptic collagen epitope within ischemic muscle suggests an important role for collagen remodeling during the active phase of ischemic limb reperfusion. PMID- 16251420 TI - Impaired rapid eye movement sleep in the Tg2576 APP murine model of Alzheimer's disease with injury to pedunculopontine cholinergic neurons. AB - Impaired rapid eye movement sleep (REMS) is commonly observed in Alzheimer's disease, suggesting injury to mesopontine cholinergic neurons. We sought to determine whether abnormal beta-amyloid peptides impair REMS and injure mesopontine cholinergic neurons in transgenic (hAPP695.SWE) mice (Tg2576) that model brain amyloid pathologies. Tg2576 mice and wild-type littermates were studied at 2, 6, and 12 months by using sleep recordings, contextual fear conditioning, and immunohistochemistry. At 2 months of age, REMS was indistinguishable by genotype but was reduced in Tg2576 mice at 6 and 12 months. Choline acetyltransferase-positive neurons in the pedunculopontine tegmentum of Tg2576 mice at 2 months evidenced activated caspase-3 immunoreactivity, and at 6 and 12 months the numbers of pedunculopontine tegmentum choline acetyltransferase positive neurons were reduced in the Tg2576 mice. Other cholinergic groups involved in REMS were unperturbed. At 12 months, Tg2576 mice demonstrated increased 3-nitrotyrosine immunoreactivity in cholinergic projection sites but not in cholinergic soma. We have identified a population of selectively compromised cholinergic neurons in young Tg2576 mice that manifest early onset REMS impairment. The differential vulnerability of these cholinergic neurons to Abeta injury provides an invaluable tool with which to understand mechanisms of sleep/wake perturbations in Alzheimer's disease. PMID- 16251421 TI - Absence of 12/15 lipoxygenase reduces brain oxidative stress in apolipoprotein E deficient mice. AB - The enzyme 12/15 lipoxygenase (12/15LO) has been implicated in the oxidative modification of lipoproteins and phospholipids in vivo. In addition, mice deficient in apolipoprotein E (ApoE-/-) are characterized by spontaneous hypercholesterolemia and a systemic increase in oxidative stress. Whereas the absence of 12/15LO reduces lipid peroxidation in the plasma and urine of ApoE-/- mice, the relative contribution of this enzyme to oxidative stress in the central nervous system remains unknown. Here, we provide the first in vivo evidence that 12/15LO modulates brain oxidative stress reactions using ApoE-/- mice crossbred with 12/15LO-deficient (12/15LO-/-) mice (12/15LO-/-/ApoE-/-). In chow-fed 12 month-old 12/15LO-/-/ApoE-/- mice, the amount of brain isoprostane iPF2alpha-VI, a marker of lipid peroxidation, and carbonyls, markers of protein oxidation, were significantly reduced when compared with 12/15LO-expressing controls (12/15LO+/+/ApoE-/-). These results were observed despite the fact that cholesterol, triglyceride, and lipoprotein levels were similar to those of ApoE-/ mice. These data indicate a functional role for 12/15LO in the modulation of oxidative reactions in the central nervous system, supporting the hypothesis that inhibition of this enzymatic pathway may be a novel therapeutic target in clinical settings involving increased brain oxidative stress. PMID- 16251422 TI - In situ analysis of integrin and growth factor receptor signaling pathways in human glioblastomas suggests overlapping relationships with focal adhesion kinase activation. AB - Deregulated integrin signaling is common in cancers, including glioblastoma. Integrin binding and growth factor receptor signaling activate focal adhesion kinase (FAK) and subsequently up-regulate extracellular regulated kinases (ERK 1/2), leading to cell-cycle progression and cell migration. Most studies of this pathway have used in vitro systems or tumor lysate-based approaches. We examined these pathways primarily in situ using a panel of 30 glioblastomas and gene expression arrays, immunohistochemistry, and fluorescence in situ hybridization, emphasizing the histological distribution of molecular changes. Within individual tumors, increased expression of FAK, p-FAK, paxillin, ERK-1/2, and p-ERK-1/2 occurred in regions of elevated EGFR and/or PDGFRA expression. Moreover, FAK activation levels correlated with EGFR and PDGFRA expression, and p-FAK and EGFR expression co-localized at the single-cell level. In addition, integrin expression was enriched in EGFR/PDGFRA-overexpressing areas but was more regionally confined than FAK, p-FAK, and paxillin. Integrins beta8 and alpha5beta1 were most commonly expressed, often in a perinecrotic or perivascular pattern. Taken together, our data suggest that growth factor receptor overexpression facilitates alterations in the integrin signaling pathway. Thus, FAK may act in glioblastoma as a downstream target of growth factor signaling, with integrins enhancing the impact of such signaling in the tumor microenvironment. PMID- 16251423 TI - Rapid vessel regression, protease inhibition, and stromal normalization upon short-term vascular endothelial growth factor receptor 2 inhibition in skin carcinoma heterotransplants. AB - Vascular endothelial growth factor (VEGF) plays a key role in tumor angiogenesis, and blockade of VEGF receptor 2 (VEGFR-2), with the monoclonal antibody DC101, inhibits angiogenesis and tumor growth. To examine the short-term effects of DC101, we surface transplanted the squamous cell carcinoma cell line A5-RT3 onto nude mice. After short-term treatment with DC101, we observed rapid reduction in vascularization and reversion of the tumor phenotype. Beginning 24 hours after treatment, VEGFR-2 inhibition resulted in decreased vessel density within the tenascin-c-staining tumor-associated stroma and reduced endothelial cell proliferation. Stromal expression of matrix metalloproteinase-9 and -13 was drastically reduced 96 hours after VEGFR-2 inhibition as detected by in situ hybridization and in situ zymography. Moreover, the morphology of the tumor stroma border changed from a highly invasive carcinoma to a well-demarcated, premalignant phenotype. The latter was characterized by the appearance of a regular basement membrane in immunostaining and ultrastructural analyses. These findings suggest that VEGFR-2 inhibition by DC101 evokes very rapid reduction of preformed vessels and decreases both stromal protease expression and gelatinolytic activity, resulting in the modulation of the tumor-stroma border zone and reversion of the tumor phenotype. Thus, short-term inhibition of VEGF signaling results in complex stromal alterations with crucial consequences for the tumor phenotype. PMID- 16251424 TI - Stromal regulation of neoplastic development: age-dependent normalization of neoplastic mammary cells by mammary stroma. AB - There is mounting evidence that the stroma plays a crucial role in mammary gland carcinogenesis. Here, we report that mammary gland stroma from mature and multiparous rats prevents neoplastic development and encourages normal ductal growth of grafted epithelial cancer cells. Fifty thousand epithelial cancer cells were injected into the cleared fat pads of virgin hosts at 24, 52, 80, and 150 days of age and of hosts that had undergone two cycles of pregnancy, lactation, and involution. Six months after inoculation, tumor incidence was 75%, 100%, 50%, and 18.2% in 24-, 52-, 80-, and 150-day-old virgin rats, respectively, and 0% in the twice-parous animals. Most remarkably, these neoplastic cells appeared to form normal ducts in all hosts-Ha-ras-1 mutation served as a marker to identify the tumor origin of the outgrowths. The tumor development pattern suggests a parallel to the phenomenon of age- and reproductive state-dependent susceptibility and resistance to chemical carcinogens. As susceptibility to carcinogenesis decreases, the ability of the stroma to reprogram neoplastic epithelial cells increases. Thus, the neoplastic phenotype is context-dependent, and it therefore offers the intriguing possibility that the process of carcinogenesis is amenable to normalization or cure once the mechanisms of stroma mediated normalization are elucidated and manipulated. PMID- 16251425 TI - Simultaneous expression of caveolin-1 and E-cadherin in ovarian carcinoma cells stabilizes adherens junctions through inhibition of src-related kinases. AB - Cadherin-mediated adhesion plays an important role in maintaining cell-cell contacts and reducing tumor metastasis. However, neo-expression of E-cadherin in ovarian carcinoma does not prevent the release and spread of cells from the primary tumor. Because caveolin-1 is down-regulated concomitantly with E-cad expression, we investigated whether the stability of adherens junctions in ovarian carcinoma was affected by caveolin-1 expression. We used IGROV1 cells transfected with caveolin-1 (IGtC3), mock-transfected control cells (IGtM87), and SKOV3 cells that endogenously express caveolin-1. Simultaneous expression of caveolin-1 and E-cadherin favored membrane distribution of E-cadherin and its associated catenin (p120ctn), even when caveolin-1 was only focally associated with adherens junctions. Silencing of caveolin-1 induced intracellular E-cadherin redistribution in IGtC3 and SKOV3 cells. Treatment with the specific src kinase inhibitor PP1 increased E-cadherin expression in IGtM87 and SKOV3 cells and enhanced membrane localization of both E-cadherin and p120ctn. However, PP1 could not completely reverse the detrimental effects on cell-cell adhesion induced by Ca2+ depletion in IGtM87 cells. Together, our data suggest that caveolin-1 expression indirectly promotes cell-cell adhesion in ovarian carcinoma cells by a mechanism involving inhibition of src-related kinases. Thus, down-regulation or loss of caveolin-1 might contribute significantly to the spread of tumor cells from the primary tumor. PMID- 16251426 TI - Angiotensin II activates matrix metalloproteinase type II and mimics age associated carotid arterial remodeling in young rats. AB - Increased angiotensin II (Ang II), matrix metalloproteinase type II (MMP2), and sympathetic activity accompany age-associated arterial remodeling. To analyze this relationship, we infused a low subpressor dose of Ang II into young (8 months old) rats. This increased carotid arterial MMP2 transcription, translation, and activation, as well as transforming growth factor-beta1 activity and collagen deposition. A higher Ang II concentration, which increased arterial pressure to that of old (30 months old) untreated rats, produced carotid media thickening and intima infiltration by vascular smooth muscle cells (VSMCs). Ex vivo, Ang II increased MMP2 activity in carotid rings from young rats to that of untreated old rats. Ang II also increased the ability of early passage VSMCs from young rats to invade a synthetic basement membrane, similar to that of untreated VSMCs from old rats. The MMP inhibitor GM6001 and the AT1 receptor antagonist Losartan inhibited these effects. The alpha-adrenoreceptor agonist phenylephrine increased arterial Ang II protein, causing MMP2 activation and intima and media thickening. Exposure of young VSMCs to phenylephrine in vitro increased Ang II protein and MMP2 activity to the levels of old VSMCs; Losartan abolished these effects. Thus, Ang II-induced effects on MMP2, transforming growth factor-beta1, collagen, and VSMCs are central to the arterial remodeling that accompanies advancing age. PMID- 16251427 TI - Mechanisms of abruption-induced premature rupture of the fetal membranes: thrombin-enhanced interleukin-8 expression in term decidua. AB - Recent evidence has linked preterm premature rupture of the fetal membranes (PPROM) to placental abruption. Because neutrophils are a rich source of proteases that can degrade extracellular matrix in abruption-associated PPROM, we examined whether decidual neutrophil infiltration complicates abruption associated PPROM. Accordingly, immunostaining for the neutrophil marker CD15 was performed in placentas obtained after overt abruption (decidual hemorrhage) with or without PPROM and in control placentas. Abruptions were associated with a marked decidual neutrophil infiltration that peaked after PPROM, whereas decidua from gestational age-matched controls were virtually devoid of neutrophils. Neutrophil infiltrates co-localized with fibrin deposition. Because abruptions elicit intense decidua-enhanced thrombin production, we examined the regulation of abruption-induced neutrophil infiltration. Expression of the primary neutrophil chemoattractant interleukin-8 (IL-8) was evaluated in leukocyte-free term decidual cells incubated with estradiol (E2; control) or with E2+medroxyprogesterone acetate (to mimic pregnancy)+/-thrombin. After 24 hours, enzyme-linked immunosorbent assay measurements indicated that thrombin (0.1 to 2.5 U/ml) elicited a dose-dependent elevation in secreted IL-8 (P<0.05) with 2.5 U/ml of thrombin increasing IL-8 levels by >14-fold in E2 and E2+medroxyprogesterone incubations. Results were validated by Western blot and quantitative reverse transcriptase-polymerase chain reaction. In summary, thrombin-enhanced IL-8 expression in term decidual cells may explain how abruption-associated PPROM promotes decidual neutrophil infiltration. PMID- 16251428 TI - Vascular endothelial growth factor expression in the retinal pigment epithelium is essential for choriocapillaris development and visual function. AB - The choroid in the eye provides vascular support for the retinal pigment epithelium (RPE) and the photoreceptors. Vascular endothelial growth factor (VEGF) derived from the RPE has been implicated in the physiological regulation of the choroidal vasculature, and overexpression of VEGF in this epithelium has been considered an important factor in the pathogenesis of choroidal neovascularization in age-related macular degeneration. Here, we demonstrate that RPE-derived VEGF is essential for choriocapillaris development. Conditional inactivation of VEGF expression in the RPE (in VEGFrpe-/- mice) results in the absence of choriocapillaris, occurrence of microphthalmia, and the loss of visual function. Severe abnormalities of RPE cells are already observed when VEGF expression in the RPE is only reduced (in VEGFrpe+/- mice), despite the formation of choroidal vessels at these VEGF levels. Finally, using Hif1arpe-/- mice we demonstrate that these roles of VEGF are not dependent on hypoxia-inducible factor-1alpha-mediated transcriptional regulation of VEGF expression in the RPE. Thus, hypoxia-inducible factor-1alpha-independent expression of VEGF is essential for choroid development. PMID- 16251429 TI - A TOP at the Synapse. PMID- 16251430 TI - Regulation of Kv7 (KCNQ) K+ channel open probability by phosphatidylinositol 4,5 bisphosphate. AB - Voltage-gated Kv7 (KCNQ) channels underlie important K+ currents, including the neuronal M current, and are thought to be sensitive to membrane phosphatidylinositol 4,5-bisphosphate (PIP2) and PIP2 depletion to underlie muscarinic receptor inhibition. We studied regulation of Kv7.2-7.4 channels by PIP2 in Chinese hamster ovary (CHO) cells using single-channel and whole-cell patch clamp and biochemical analysis. Maximal open probabilities (Po) of Kv7.2 Kv7.4 homomultimers and of Kv7.2/7.3 heteromultimers were found to be strongly dependent on the [diC8-PIP2] applied to inside-out patches, with differential apparent affinities that correlate with their maximal Po in on-cell mode. Unitary conductance was not affected by PIP2. Raising tonic [PIP2] by coexpression of phosphatidylinositol (4)5-kinase increased the maximal Po of both Kv7.2 and Kv7.2/7.3 channels studied in on-cell patches and increased whole-cell Kv7.2, but not Kv7.3, current amplitudes. In cells coexpressed with muscarinic M1 receptors, bath application of muscarinic agonist reduced the maximal Po of Kv7.2/7.3 channels isolated in on-cell patches. Coexpression of a PIP2 sequestering construct moderately reduced whole-cell Kv7.2/7.3 currents, and coexpression of a construct containing a PIP2 phosphatase nearly abolished them. Finally, biochemical analysis of anionic phospholipids in CHO cells stably expressing M1 receptors shows that PIP2 and PIP are nearly depleted 1 min after muscarinic stimulation, with an unexpected rebound after 10 min. These results strongly support the direct regulation of Kv7 channels by PIP2 and its depletion as the mechanism of muscarinic suppression of M channels. Divergent apparent affinities of Kv7.2-7.4 channels for PIP2 may underlie their highly differential maximal Po observed in cell-attached patches. PMID- 16251431 TI - Alpha7 neuronal nicotinic acetylcholine receptors are negatively regulated by tyrosine phosphorylation and Src-family kinases. AB - Nicotine, a component of tobacco, is highly addictive but possesses beneficial properties such as cognitive improvements and memory maintenance. Involved in these processes is the neuronal nicotinic acetylcholine receptor (nAChR) alpha7, whose activation triggers depolarization, intracellular signaling cascades, and synaptic plasticity underlying addiction and cognition. It is therefore important to investigate intracellular mechanisms by which a cell regulates alpha7 nAChR activity. We have examined the role of phosphorylation by combining molecular biology, biochemistry, and electrophysiology in SH-SY5Y neuroblastoma cells, Xenopus oocytes, rat hippocampal interneurons, and neurons from the supraoptic nucleus, and we found tyrosine phosphorylation of alpha7 nAChRs. Tyrosine kinase inhibition by genistein decreased alpha7 nAChR phosphorylation but strongly increased acetylcholine-evoked currents, whereas tyrosine phosphatase inhibition by pervanadate produced opposite effects. Src-family kinases (SFKs) directly interacted with the cytoplasmic loop of alpha7 nAChRs and phosphorylated the receptors at the plasma membrane. SFK inhibition by PP2 [4-amino-5-(4 chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine] or SU6656 (2,3-dihydro-N,N dimethyl-2-oxo-3-[(4,5,6,7-tetrahydro-1H-indol-2-yl)methylene]-1H-indole-5 sulfonamide) increased alpha7 nAChR-mediated responses, whereas expression of active Src reduced alpha7 nAChR activity. Mutant alpha7 nAChRs lacking cytoplasmic loop tyrosine residues because of alanine replacement of Tyr-386 and Tyr-442 were more active than wild-type receptors and insensitive to kinase or phosphatase inhibition. Because the amount of surface alpha7 receptors was not affected by kinase or phosphatase inhibitors, these data show that functional properties of alpha7 nAChRs depend on the tyrosine phosphorylation status of the receptor and are the result of a balance between SFKs and tyrosine phosphatases. These findings reveal novel regulatory mechanisms that may help to understand nicotinic receptor-dependent plasticity, addiction, and pathology. PMID- 16251432 TI - Brainstem timing: implications for cortical processing and literacy. AB - The search for a unique biological marker of language-based learning disabilities has so far yielded inconclusive findings. Previous studies have shown a plethora of auditory processing deficits in learning disabilities at both the perceptual and physiological levels. In this study, we investigated the association among brainstem timing, cortical processing of stimulus differences, and literacy skills. To that end, brainstem timing and cortical sensitivity to acoustic change [mismatch negativity (MMN)] were measured in a group of children with learning disabilities and normal-learning children. The learning-disabled (LD) group was further divided into two subgroups with normal and abnormal brainstem timing. MMNs, literacy, and cognitive abilities were compared among the three groups. LD individuals with abnormal brainstem timing were more likely to show reduced processing of acoustic change at the cortical level compared with both normal learning individuals and LD individuals with normal brainstem timing. This group was also characterized by a more severe form of learning disability manifested by poorer reading, listening comprehension, and general cognitive ability. We conclude that abnormal brainstem timing in learning disabilities is related to higher incidence of reduced cortical sensitivity to acoustic change and to deficient literacy skills. These findings suggest that abnormal brainstem timing may serve as a reliable marker of a subgroup of individuals with learning disabilities. They also suggest that faulty mechanisms of neural timing at the brainstem may be the biological basis of malfunction in this group. PMID- 16251433 TI - Postsynaptically synthesized prostaglandin E2 (PGE2) modulates hippocampal synaptic transmission via a presynaptic PGE2 EP2 receptor. AB - Increasing evidence suggests that cyclooxygenase-2 (COX-2) is involved in synaptic transmission and plasticity, and prostaglandin E2 (PGE2) is a key molecule in COX-2-meduated synaptic modification. However, the precise mechanisms, in particular, which subtypes of PGE2 receptors (EPs) mediate the PGE2-induced synaptic response, are not clear. Recently, we demonstrated that EPs are expressed heterogeneously in the hippocampus, and EP2/4 are mainly expressed in presynaptic terminals. Here, we report that PGE2 increased synaptic stimulus evoked amplitudes of EPSPs in hippocampal slices and frequency of miniature EPSCs (mEPSCs) in hippocampal neurons in culture. These actions were mimicked by an EP2 agonist and attenuated by protein kinase A inhibitors. Decrease of EP2 expression through silencing the EP2 gene eliminated PGE2-induced increase of the frequency of mEPSCs. COX-2 and microsomal PGE synthase-1 (mPGES-1) and mPGES-2 are present in postsynaptic dendritic spines, because they are colocalized with PSD-95 (postsynaptic density-95), a postsynaptic marker. In addition, the frequency of mEPSCs was enhanced in neurons pretreated with interleukin-1beta or lipopolysaccharide, which elevated expression of COX-2 and mPGES-1 and produced PGE2, and this enhancement was inhibited by a COX-2 inhibitor that inhibited production of PGE2. Our results suggest that PGE2 synthesized by postsynaptically localized COX-2 functions as a retrograde messenger in hippocampal synaptic signaling via a presynaptic EP2 receptor. PMID- 16251434 TI - Impaired regulation of thalamic pacemaker channels through an imbalance of subunit expression in absence epilepsy. AB - The role of hyperpolarization-activated, cyclic nucleotide-modulated (HCN) channel isoforms and hyperpolarization-activated cation current (Ih) for seizure related burst firing in thalamocortical (TC) neurons was investigated in a rat genetic model of absence epilepsy [Wistar Albino Glaxo rats, bred in Rijswijk (WAG/Rij)]. Burst discharges in TC neurons locked to seizure activity in vivo were prolonged during blockade of Ih by Cs+ and ZD7288 (4-ethylphenylamino-1,2 dimethyl-6-methylaminopyrimidinium chloride). In vitro analyses revealed a hyperpolarizing shift of half-maximal Ih activation (Vh) in WAG/Rij (Vh = -93.2 mV) compared with nonepileptic controls [August x Copenhagen-Irish (ACI) (Vh = 88.0 mV)]. This effect is explained by a shift of the responsiveness of Ih to cAMP toward higher concentrations in TC neurons from WAG/Rij, as revealed by application of 8-bromo-cAMP and the phosphodiesterase inhibitor IBMX. During blockade of adenylyl cyclase activity, Ih activation was similar in the two strains, whereas the difference in cAMP responsiveness persisted, thereby voting against different ambient cAMP levels between strains. Increasing the intracellular cAMP level and shifting Ih activation led to a change from burst to tonic firing mode in WAG/Rij but not in ACI rats. Furthermore, HCN1 expression was significantly increased on mRNA and protein levels, with no changes in HCN2-4 expression. In conclusion, there is an increase in HCN1 expression in the epileptic thalamus, associated with a decrease in cAMP responsiveness of Ih in TC neurons and resulting impairment to control the shift from burst to tonic firing, which, in turn, will prolong burst activity after recruitment of Ih during absence seizures. PMID- 16251435 TI - Role of hippocampal Cav1.2 Ca2+ channels in NMDA receptor-independent synaptic plasticity and spatial memory. AB - Current knowledge about the molecular mechanisms of NMDA receptor (NMDAR) independent long-term potentiation (LTP) in the hippocampus and its function for memory formation in the behaving animal is limited. NMDAR-independent LTP in the CA1 region is thought to require activity of postsynaptic L-type voltage dependent Ca2+ channels (Cav1.x), but the underlying channel isoform remains unknown. We evaluated the function of the Cav1.2 L-type Ca2+ channel for spatial learning, synaptic plasticity, and triggering of learning-associated biochemical processes using a mouse line with an inactivation of the CACNA1C (Cav1.2) gene in the hippocampus and neocortex (Cav1.2(HCKO)). This model shows (1) a selective loss of protein synthesis-dependent NMDAR-independent Schaffer collateral/CA1 late-phase LTP (L-LTP), (2) a severe impairment of hippocampus-dependent spatial memory, and (3) decreased activation of the mitogen-activated protein kinase (MAPK) pathway and reduced cAMP response element (CRE)-dependent transcription in CA1 pyramidal neurons. Our results provide strong evidence for a role of L-type Ca2+ channel-dependent, NMDAR-independent hippocampal L-LTP in the formation of spatial memory in the behaving animal and for a function of the MAPK/CREB (CRE binding protein) signaling cascade in linking Cav1.2 channel-mediated Ca2+ influx to either process. PMID- 16251436 TI - Transgenic expression of a dominant-negative ASIC3 subunit leads to increased sensitivity to mechanical and inflammatory stimuli. AB - Molecular and behavioral evidence suggests that acid-sensing ion channels (ASICs) contribute to pain processing, but an understanding of their precise role remains elusive. Existing ASIC knock-out mouse experiments are complicated by the heteromultimerization of ASIC subunits. Therefore, we have generated transgenic mice that express a dominant-negative form of the ASIC3 subunit that inactivates all native neuronal ASIC-like currents by oligomerization. Using whole-cell patch clamp recordings, we examined the response properties of acutely isolated dorsal root ganglion neurons to protons (pH 5.0). We found that whereas 33% of the proton-responsive neurons from wild-type mice exhibited an ASIC-like transient response, none of the neurons from the transgenic mice exhibited a transient inward current. Capsaicin-evoked responses mediated by the TRPV1 receptor were unaltered in transgenic mice. Adult male wild-type and transgenic mice were subjected to a battery of behavioral nociceptive assays, including tests of thermal, mechanical, chemical/inflammatory, and muscle pain. The two genotypes were equally sensitive to thermal pain and to thermal hypersensitivity after inflammation. Compared with wild types, however, transgenic mice were more sensitive to a number of modalities, including mechanical pain (von Frey test, tail-clip test), chemical/inflammatory pain (formalin test, 0.6% acetic acid writhing test), mechanical hypersensitivity after zymosan inflammation, and mechanical hypersensitivity after intramuscular injection of hypotonic saline. These data reinforce the hypothesis that ASICs are involved in both mechanical and inflammatory pain, although the increased sensitivity of transgenic mice renders it unlikely that they are direct transducers of nociceptive stimuli. PMID- 16251437 TI - Cooperative glutamatergic and cholinergic mechanisms generate short-term modifications of synaptic effectiveness in prepositus hypoglossi neurons. AB - To maintain horizontal eye position on a visual target after a saccade, extraocular motoneurons need a persistent (tonic) neural activity, called "eye position signal," generated by prepositus hypoglossi (PH) neurons. We have shown previously in vitro and in vivo that this neural activity depends, among others mechanisms, on the interplay of glutamatergic transmission and cholinergic synaptically triggered depolarization. Here, we used rat sagittal brainstem slices, including PH nucleus and paramedian pontine reticular formation (PPRF). We made intracellular recordings of PH neurons and studied their synaptic activation from PPRF neurons. Train stimulation of the PPRF area evoked a cholinergic-sustained depolarization of PH neurons that outlasted the stimulus. EPSPs evoked in PH neurons by single pulses applied to the PPRF presented a short term potentiation (STP) after train stimulation. APV (an NMDA-receptor blocker) or chelerythrine (a protein kinase-C inhibitor) had no effect on the sustained depolarization, but they did block the evoked STP, whereas pirenzepine (an M1 muscarinic antagonist) blocked both the sustained depolarization and the STP of PH neurons. Thus, electrical stimulation of the PPRF area activates both glutamatergic and cholinergic axons terminating in the PH nucleus, the latter producing a sustained depolarization probably involved in the genesis of the persistent neural activity required for eye fixation. M1-receptor activation seems to evoke a STP of PH neurons via NMDA receptors. Such STP could be needed for the stabilization of the neural network involved in the generation of position signals necessary for eye fixation after a saccade. PMID- 16251438 TI - The temporal impulse response underlying saccadic decisions. AB - Models of perceptual decision making often assume that sensory evidence is accumulated over time in favor of the various possible decisions, until the evidence in favor of one of them outweighs the evidence for the others. Saccadic eye movements are among the most frequent perceptual decisions that the human brain performs. We used stochastic visual stimuli to identify the temporal impulse response underlying saccadic eye movement decisions. Observers performed a contrast search task, with temporal variability in the visual signals. In experiment 1, we derived the temporal filter observers used to integrate the visual information. The integration window was restricted to the first approximately 100 ms after display onset. In experiment 2, we showed that observers cannot perform the task if there is no useful information to distinguish the target from the distractor within this time epoch. We conclude that (1) observers did not integrate sensory evidence up to a criterion level, (2) observers did not integrate visual information up to the start of the saccadic dead time, and (3) variability in saccade latency does not correspond to variability in the visual integration period. Instead, our results support a temporal filter model of saccadic decision making. The temporal impulse response identified by our methods corresponds well with estimates of integration times of V1 output neurons. PMID- 16251439 TI - Hippocampal neurons express a calcineurin-activated adenylyl cyclase. AB - Ca2+-stimulated adenylyl cyclases are important for several forms of neuroplasticity because they couple activity-dependent Ca2+ increases to cAMP in neurons. For example, the calmodulin-stimulated adenylyl cyclases, AC1 and AC8, are required for hippocampus-dependent memory and long-lasting long-term potentiation. To identify other mechanisms for Ca2+ stimulation of adenylyl cyclases, cultured hippocampal neurons from transgenic mice lacking both AC1 and AC8 [double knock-out (DKO) mice] were analyzed for Ca2+ stimulation of intracellular cAMP. Surprisingly, neurons from DKO mice showed significant Ca2+ stimulated cAMP accumulation that was blocked by inhibitors of calcineurin [PP2B (protein phosphatase 2B)], a Ca2+-activated protein phosphatase. Analysis of cultured neurons from calcineurin(-/-) mice confirmed that hippocampal neurons exhibit a calcineurin-dependent cAMP increase, which may contribute to some forms of neuroplasticity. PMID- 16251440 TI - Neural correlates of reach errors. AB - Reach errors may be broadly classified into errors arising from unpredictable changes in target location, called target errors, and errors arising from miscalibration of internal models (e.g., when prisms alter visual feedback or a force field alters limb dynamics), called execution errors. Execution errors may be caused by miscalibration of dynamics (e.g., when a force field alters limb dynamics) or by miscalibration of kinematics (e.g., when prisms alter visual feedback). Although all types of errors lead to similar on-line corrections, we found that the motor system showed strong trial-by-trial adaptation in response to random execution errors but not in response to random target errors. We used functional magnetic resonance imaging and a compatible robot to study brain regions involved in processing each kind of error. Both kinematic and dynamic execution errors activated regions along the central and the postcentral sulci and in lobules V, VI, and VIII of the cerebellum, making these areas possible sites of plastic changes in internal models for reaching. Only activity related to kinematic errors extended into parietal area 5. These results are inconsistent with the idea that kinematics and dynamics of reaching are computed in separate neural entities. In contrast, only target errors caused increased activity in the striatum and the posterior superior parietal lobule. The cerebellum and motor cortex were as strongly activated as with execution errors. These findings indicate a neural and behavioral dissociation between errors that lead to switching of behavioral goals and errors that lead to adaptation of internal models of limb dynamics and kinematics. PMID- 16251441 TI - Dysfunction of the cholesterol biosynthetic pathway in Huntington's disease. AB - The expansion of a polyglutamine tract in the ubiquitously expressed huntingtin protein causes Huntington's disease (HD), a dominantly inherited neurodegenerative disease. We show that the activity of the cholesterol biosynthetic pathway is altered in HD. In particular, the transcription of key genes of the cholesterol biosynthetic pathway is severely affected in vivo in brain tissue from HD mice and in human postmortem striatal and cortical tissue; this molecular dysfunction is biologically relevant because cholesterol biosynthesis is reduced in cultured human HD cells, and total cholesterol mass is significantly decreased in the CNS of HD mice and in brain-derived ST14A cells in which the expression of mutant huntingtin has been turned on. The transcription of the genes of the cholesterol biosynthetic pathway is regulated via the activity of sterol regulatory element-binding proteins (SREBPs), and we found an approximately 50% reduction in the amount of the active nuclear form of SREBP in HD cells and mouse brain tissue. As a consequence, mutant huntingtin reduces the transactivation of an SRE-luciferase construct even under conditions of SREBP overexpression or in the presence of an exogenous N-terminal active form of SREBP. Finally, the addition of exogenous cholesterol to striatal neurons expressing mutant huntingtin prevents their death in a dose-dependent manner. We conclude that the cholesterol biosynthetic pathway is impaired in HD cells, mice, and human subjects, and that the search for HD therapies should also consider cholesterol levels as both a potential target and disease biomarker. PMID- 16251443 TI - Identification of nicotinic acetylcholine receptor recycling and its role in maintaining receptor density at the neuromuscular junction in vivo. AB - In the CNS, receptor recycling is critical for synaptic plasticity; however, the recycling of receptors has never been observed at peripheral synapses. Using a novel imaging technique, we show here that nicotinic acetylcholine receptors (AChRs) recycle into the postsynaptic membrane of the neuromuscular junction. By sequentially labeling AChRs with biotin-bungarotoxin and streptavidin-fluorophore conjugates, we were able to distinguish recycled, preexisting, and new receptor pools at synapses in living mice. Time-lapse imaging revealed that recycled AChRs were incorporated into the synapse within hours of initial labeling, and their numbers increased with time. At fully functional synapses, AChR recycling was robust and comparable in magnitude with the insertion of newly synthesized receptors, whereas chronic synaptic activity blockade nearly abolished receptor recycling. Finally, using the same sequential labeling method, we found that acetylcholinesterase, another synaptic component, does not recycle. These results identify an activity-dependent AChR-recycling mechanism that enables the regulation of receptor density, which could lead to rapid alterations in synaptic efficacy. PMID- 16251442 TI - Vascular development of the brain requires beta8 integrin expression in the neuroepithelium. AB - We showed previously that loss of the integrin beta8 subunit, which forms alphavbeta8 heterodimers, results in abnormal vascular development in the yolk sac, placenta, and brain. Animals lacking the integrin beta8 (itgbeta8) gene die either at midgestation, because of insufficient vascularization of the placenta and yolk sac, or shortly after birth with severe intracerebral hemorrhage. To specifically focus on the role of integrins containing the beta8 subunit in the brain, and to avoid early lethalities, we used a targeted deletion strategy to delete itgbeta8 only from cell types within the brain. Ablating itgbeta8 from vascular endothelial cells or from migrating neurons did not result in cerebral hemorrhage. Targeted deletion of itgbeta8 from the neuroepithelium, however, resulted in bilateral hemorrhage at postnatal day 0, although the phenotype was less severe than in itgbeta8-null animals. Newborn mice lacking itgbeta8 from the neuroepithelium had hemorrhages in the cortex, ganglionic eminence, and thalamus, as well as abnormal vascular morphogenesis, and disorganized glia. Interestingly, adult mice lacking itgbeta8 from cells derived from the neuroepithelium did not show signs of hemorrhage. We propose that defective association between vascular endothelial cells and glia lacking itgbeta8 is responsible for the leaky vasculature seen during development but that an unidentified compensatory mechanism repairs the vasculature after birth. PMID- 16251444 TI - Na+-dependent sources of intra-axonal Ca2+ release in rat optic nerve during in vitro chemical ischemia. AB - The contribution of intracellular stores to axonal Ca2+ overload during chemical ischemia in vitro was examined by confocal microscopy. Ca2+ accumulation was measured by fluo-4 dextran (low-affinity dye, KD approximately 4 microM) or by Oregon Green 488 BAPTA-1 dextran (highaffinity dye, KD approximately 450 nM). Axonal Na+ was measured using CoroNa Green. Ischemia in CSF containing 2 mM Ca2+ caused an approximately 3.5-fold increase in fluo-4 emission after 30 min, indicating a large axonal Ca2+ rise well into the micromolar range. Axonal Na+ accumulation was enhanced by veratridine and reduced, but not abolished, by TTX. Ischemia in Ca2+-free (plus BAPTA) perfusate resulted in a smaller but consistent Ca2+ increase monitored by Oregon Green 488 BAPTA-1, indicating release from intracellular sources. This release was eliminated in large part when Na+ influx was reduced by replacement with N-methyl-D-glucamine (NMDG+; even in depolarizing high K+ perfusate), Li+, or by the application of TTX and significantly increased by veratridine. Intracellular release also was reduced significantly by neomycin or 1-(6-[(17beta-methoxyestra-1,3,5 [10]-trien-17-yl) amino] hexyl)-1H-pyrrole 2,5-dione (U73122 [GenBank]) (phospholipase C inhibitors), heparin [inositol trisphosphate (IP3) receptor blocker], or 7-chloro-5-(2-chlorophenyl)-1,5-dihydro 4,1-benzothiazepin-2(3H)-one (CGP37157; mitochondrial Na+/Ca2+ exchange inhibitor) as well as ryanodine. Combining CGP37157 with U73122 [GenBank] or heparin decreased the response more than either agent alone and significantly improved electrophysiological recovery. Our conclusion is that intra-axonal Ca2+ release during ischemia in rat optic nerve is mainly dependent on Na+ influx. This Na+ accumulation stimulates three distinct intra-axonal sources of Ca2+: (1) the mitochondrial Na+/Ca2+ exchanger driven in the Na+ import/Ca2+ export mode, (2) positive modulation of ryanodine receptors, and (3) promotion of IP3 generation by phospholipase C. PMID- 16251445 TI - Deterministic multiplicative gain control with active dendrites. AB - Multiplicative gain control is a vital component of many theoretical analyses of neural computations, conferring the ability to scale neuronal firing rate in response to synaptic inputs. Many theories of gain control in single cells have used precisely balanced noisy inputs. Such noisy inputs can degrade signal processing. We demonstrate a deterministic method for the control of gain without the use of noise. We show that a depolarizing afterpotential (DAP), arising from active dendritic spike backpropagation, leads to a multiplicative increase in gain. Reduction of DAP amplitude by dendritic inhibition dilutes the multiplicative effect, allowing for divisive scaling of the firing rate. In contrast, somatic inhibition acts in a subtractive manner, allowing spatially distinct inhibitory inputs to perform distinct computations. The simplicity of this mechanism and the ubiquity of its elementary components suggest that many cell types have the potential to display a dendritic division of neuronal output. PMID- 16251446 TI - Two-dimensional time coding in the auditory brainstem. AB - Avian nucleus magnocellularis (NM) spikes provide a temporal code representing sound arrival times to downstream neurons that compute sound source location. NM cells act as high-pass filters by responding only to discrete synaptic events while ignoring temporally summed EPSPs. This high degree of input selectivity insures that each output spike from NM unambiguously represents inputs that contain precise temporal information. However, we lack a quantitative description of the computation performed by NM cells. A powerful model for predicting output firing rate given an arbitrary current input is given by a linear/nonlinear cascade: the stimulus is compared with a known relevant feature by linear filtering, and based on that comparison, a nonlinear function predicts the firing response. Spike-triggered covariance analysis allows us to determine a generalization of this model in which firing depends on more than one spike triggering feature or stimulus dimension. We found two current features relevant for NM spike generation; the most important simply smooths the current on short time scales, whereas the second confers sensitivity to rapid changes. A model based on these two features captured more mutual information between current and spikes than a model based on a single feature. We used this analysis to characterize the changes in the computation brought about by pharmacological manipulation of the biophysical properties of the neurons. Blockage of low threshold voltage-gated potassium channels selectively eliminated the requirement for the second stimulus feature, generalizing our understanding of input selectivity by NM cells. This study demonstrates the power of covariance analysis for investigating single neuron computation. PMID- 16251447 TI - The p75 neurotrophin receptor negatively modulates dendrite complexity and spine density in hippocampal neurons. AB - The correlation between functional and structural neuronal plasticity is by now well documented. However, the molecular mechanisms translating patterns of neuronal activity into specific changes in the structure of neurons remain unclear. Neurotrophins can be released in an activity-dependent manner, and they are capable of controlling both neuronal morphology and functional synaptic changes. They are thus attractive molecules to be studied in the context of synaptic plasticity. In the CNS, most of the work so far has focused on the role of BDNF and of its tyrosine kinase B receptor (TrkB), but relatively little is known about the function of the pan-neurotrophin receptor p75NTR. In this study, we show in loss-of-function experiments that postnatal hippocampal pyramidal cells in two mutant lines of p75NTR have a higher spine density and greater dendritic complexity than wild-type (WT) mice. Conversely, in a gain-of-function approach, p75NTR overexpression in WT neurons significantly reduces dendritic complexity, as well as spine density in all dendritic compartments. These results show that p75NTR negatively modulates dendritic morphology in adult hippocampal pyramidal neurons and documents a new case of functional antagonism between Trk and p75NTR signaling. PMID- 16251448 TI - Protease-activated receptor-1 and platelet-derived growth factor in spinal cord neurons are implicated in neuropathic pain after nerve injury. AB - Recently, it has been reported that both thrombin-sensitive protease-activated receptor 1 (PAR-1) and platelet-derived growth factor (PDGF) are present not only in platelets, but also in the CNS, which indicates that they have various physiological functions. In this study, we evaluated whether PAR-1/PDGF in the spinal cord could contribute to the development of a neuropathic pain-like state in mice. Thermal hyperalgesia and tactile allodynia induced by sciatic nerve ligation were significantly suppressed by repeated intrathecal injection of hirudin, which is characterized as a specific and potent thrombin inhibitor. Furthermore, a single intrathecal injection of thrombin produced long-lasting hyperalgesia and allodynia, and these effects were also inhibited by hirudin in normal mice. In nerveligated mice, the increase in the binding of [35S]GTPgammaS to membranes of the spinal cord induced by thrombin and PAR-1-like immunoreactivity (IR) in the spinal cord were each greater than those in sham operated mice. Thermal hyperalgesia and tactile allodynia induced by sciatic nerve ligation were also suppressed by repeated intrathecal injection of either the PDGF alpha receptor (PDGFRalpha)/Fc chimera protein or the PDGFR-dependent tyrosine kinase inhibitor AG17 [(3,5-di-tert-butyl-4-hydroxybenzylidene) malononitrile]. Moreover, thermal hyperalgesia and tactile allodynia induced by thrombin in normal mice were virtually eliminated by intrathecal pretreatment with PDGFRalpha/Fc. In immunohistochemical studies, PAR-1-like IR-positive cells in the spinal dorsal horn were mostly colocated on PDGF-like IR-positive neuronal cells. These data provide novel evidence that PAR-1 and PDGF-A-mediated signaling pathway within spinal cord neurons may be directly implicated in neuropathic pain after nerve injury in mice. PMID- 16251449 TI - Tracking the fear engram: the lateral amygdala is an essential locus of fear memory storage. AB - Although it is believed that different types of memories are localized in discreet regions of the brain, concrete experimental evidence of the existence of such engrams is often elusive. Despite being one of the best characterized memory systems of the brain, the question of where fear memories are localized in the brain remains a hotly debated issue. Here, we combine site-specific behavioral pharmacology with multisite electrophysiological recording techniques to show that the lateral nucleus of the amygdala, long thought to be critical for the acquisition of fear memories, is also an essential locus of fear memory storage. PMID- 16251450 TI - Multiple and plastic receptors mediate tonic GABAA receptor currents in the hippocampus. AB - Persistent activation of GABAA receptors by extracellular GABA (tonic inhibition) plays a critical role in signal processing and network excitability in the brain. In hippocampal principal cells, tonic inhibition has been reported to be mediated by alpha5-subunit-containing GABAA receptors (alpha5GABAARs). Pharmacological or genetic disruption of these receptors improves cognitive performance, suggesting that tonic inhibition has an adverse effect on information processing. Here, we show that alpha5GABAARs contribute to tonic currents in pyramidal cells only when ambient GABA concentrations increase (as may occur during increased brain activity). At low ambient GABA concentrations, activation of delta-subunit containing GABAA receptors predominates. In epileptic tissue, alpha5GABAARs are downregulated and no longer contribute to tonic currents under conditions of raised extracellular GABA concentrations. Under these conditions, however, the tonic current is greater in pyramidal cells from epileptic tissue than in pyramidal cells from nonepileptic tissue, implying substitution of alpha5GABAARs by other GABAA receptor subtypes. These results reveal multiple components of tonic GABAA receptor-mediated conductance that are activated by low GABA concentrations. The relative contribution of these components changes after the induction of epilepsy, implying an adaptive plasticity of the tonic current in the presence of spontaneous seizures. PMID- 16251451 TI - Channel noise is essential for perithreshold oscillations in entorhinal stellate neurons. AB - Previous experimental and computational work (for review, see White et al., 2000) has suggested that channel noise, generated by the stochastic flicker of voltage gated ion channels, can be a major contributor to electrical membrane noise in neurons. In spiny stellate neurons of the entorhinal cortex, we remove the primary source of channel noise by pharmacologically blocking the native persistent Na+ conductance. Via the dynamic-clamp technique (Robinson and Kawai, 1993; Sharp et al., 1993), we then introduce virtual persistent Na+ channels into the membranes of the stellate neurons. By altering the mathematical properties of these virtual "knock-ins," we demonstrate that stochastic flicker of persistent Na+ channels is necessary for the existence of slow perithreshold oscillations that characterize stellate neurons. Channel noise also alters the ability of stellate neurons to phase lock to weak sinusoidal stimuli. These results provide the first direct demonstration that physiological levels of channel noise can produce qualitative changes in the integrative properties of neurons. PMID- 16251452 TI - Partial mitochondrial inhibition causes striatal dopamine release suppression and medium spiny neuron depolarization via H2O2 elevation, not ATP depletion. AB - Mitochondrial dysfunction is a potential causal factor in Parkinson's disease. We show here that acute exposure to the mitochondrial complex I inhibitor rotenone (30-100 nM; 30 min) causes concentration-dependent suppression of single-pulse evoked dopamine (DA) release monitored in real time with carbon-fiber microelectrodes in guinea pig striatal slices, with no effect on DA content. Suppression of DA release was prevented by the sulfonylurea glibenclamide, implicating ATP-sensitive K+ (KATP) channels; however, tissue ATP was unaltered. Because KATP channels can be activated by hydrogen peroxide (H2O2), as well as by low ATP, we examined the involvement of rotenone-enhanced H2O2 generation. Confirming an essential role for H2O2, the inhibition of DA release by rotenone was prevented by catalase, a peroxide-scavenging enzyme. Striatal H2O2 generation during rotenone exposure was examined in individual medium spiny neurons using fluorescence imaging with dichlorofluorescein (DCF). An increase in intracellular H2O2 levels followed a similar time course to that of DA release suppression and was accompanied by cell membrane depolarization, decreased input resistance, and increased excitability. Extracellular catalase markedly attenuated the increase in DCF fluorescence and prevented rotenone-induced effects on membrane properties; membrane changes were also largely prevented by flufenamic acid, a blocker of transient receptor potential (TRP) channels. Thus, partial mitochondrial inhibition can cause functional DA denervation via H2O2 and KATP channels, without DA or ATP depletion. Furthermore, amplified H2O2 levels and TRP channel activation in striatal spiny neurons indicate potential sources of damage in these cells. Overall, these novel factors could contribute to parkinsonian motor deficits and neuronal degeneration caused by mitochondrial dysfunction. PMID- 16251453 TI - In vivo magnetic resonance microimaging of individual amyloid plaques in Alzheimer's transgenic mice. AB - The ability to detect individual Alzheimer's amyloid plaques in vivo by magnetic resonance microimaging (MRI) should improve diagnosis and also accelerate discovery of effective therapeutic agents for Alzheimer's disease (AD). Here, we perform in vivo and ex vivo MRI on double transgenic AD mice as well as wild-type mice at varying ages and correlate these with thioflavin-S and iron staining histology. Quantitative counts of individual plaques on MRI increase with age and correlate with histologically determined plaque burden. Plaques 20 microm in diameter can be detected in AD mice as young as 3 months of age with ex vivo MRI. Plaques 35 microm in diameter can be detected by 9 months of age with in vivo MRI. In vivo MRI of individual Alzheimer's amyloid plaques provides a noninvasive estimate of plaque burden in transgenic AD mice that might be useful in assessing the efficacy of amyloid reduction therapies. PMID- 16251454 TI - A logarithmic, scale-invariant representation of speed in macaque middle temporal area accounts for speed discrimination performance. AB - Human speed discrimination thresholds follow Weber's law over a large range of reference (i.e., pedestal) speeds, that is, the just-noticeable-difference in speed scales in proportion to the reference speed. We analyzed the neural representation of speed information in macaque middle temporal visual area (MT) to determine whether this representation can account for the basic form of psychophysical data. Based on theoretical considerations, we hypothesized: (1) that the speed tuning curves of MT neurons should be bell-shaped (Gaussian) as a function of the logarithm of speed, (2) that the set of speed-tuning curves should be approximately scale-invariant, (3) that the distribution of speed preferences should be approximately uniform in log speed, and (4) that response variability should be independent of speed preference. Our quantitative analysis of data from 501 MT neurons shows that the neural representation of speed approximately obeys these constraints, with modest deviations particularly at slow speeds. We then used the MT data to predict how speed discrimination thresholds should depend on pedestal speed. The shape of this prediction matches very closely to that of human psychophysical data, accounting for constant Weber fractions over a large range of intermediate speeds as well as a marked departure from Weber's law at slow speeds. Moreover, we show that deviations of the MT representation from the above constraints are important for predicting how psychophysical thresholds depart from Weber's law at slow speeds. These findings support the notion that a logarithmic, approximately scale-invariant representation of speed in area MT limits perceptual speed discrimination. PMID- 16251455 TI - Determinants of the success of whole-genome association testing. PMID- 16251456 TI - Measures of human population structure show heterogeneity among genomic regions. AB - Estimates of genetic population structure (F(ST)) were constructed from all autosomes in two large SNP data sets. The Perlegen data set contains genotypes on approximately 1 million SNPs segregating in all three samples of Americans of African, Asian, and European descent; and the Phase I HapMap data set contains genotypes on approximately 0.6 million SNPs segregating in all four samples from specific Caucasian, Chinese, Japanese, and Yoruba populations. Substantial heterogeneity of F(ST) values was found between segments within chromosomes, although there was similarity between the two data sets. There was also substantial heterogeneity among population-specific F(ST) values, with the relative sizes of these values often changing along each chromosome. Population structure estimates are often used as indicators of natural selection, but the analyses presented here show that individual-marker estimates are too variable to be useful. There is inherent variation in these statistics because of variation in genealogy even among neutral loci, and values at pairs of loci are correlated to an extent that reflects the linkage disequilibrium between them. Furthermore, it may be that the best indications of selection will come from population specific F(ST) values rather than the usually reported population-average values. PMID- 16251457 TI - Dynamic structure of the SPANX gene cluster mapped to the prostate cancer susceptibility locus HPCX at Xq27. AB - Genetic linkage studies indicate that germline variations in a gene or genes on chromosome Xq27-28 are implicated in prostate carcinogenesis. The linkage peak of prostate cancer overlies a region of approximately 750 kb containing five SPANX genes (SPANX-A1, -A2, -B, -C, and -D) encoding sperm proteins associated with the nucleus; their expression was also detected in a variety of cancers. SPANX genes are >95% identical and reside within large segmental duplications (SDs) with a high level of similarity, which confounds mutational analysis of this gene family by routine PCR methods. In this work, we applied transformation-associated recombination cloning (TAR) in yeast to characterize individual SPANX genes from prostate cancer patients showing linkage to Xq27-28 and unaffected controls. Analysis of genomic TAR clones revealed a dynamic nature of the replicated region of linkage. Both frequent gene deletion/duplication and homology-based sequence transfer events were identified within the region and were presumably caused by recombinational interactions between SDs harboring the SPANX genes. These interactions contribute to diversity of the SPANX coding regions in humans. We speculate that the predisposition to prostate cancer in X-linked families is an example of a genomic disease caused by a specific architecture of the SPANX gene cluster. PMID- 16251458 TI - Traffic of genetic information between segmental duplications flanking the typical 22q11.2 deletion in velo-cardio-facial syndrome/DiGeorge syndrome. AB - Velo-cardio-facial syndrome/DiGeorge syndrome results from unequal crossing-over events between two 240-kb low-copy repeats termed LCR22 (LCR22-2 and LCR22-4) on Chromosome 22q11.2, comprised of modules, each of which are >99% identical in sequence. To delineate regions in the LCR22s that might contain hotspots for 22q11.2 rearrangements, we scanned the interval for increased rates of recombination with the hypothesis that these regions might be more prone to breakage. We generated an algorithm to detect sites of altered recombination by searching for single nucleotide polymorphic positions in BAC clones from different libraries mapped to LCR22-2 and LCR22-4. This method distinguishes single nucleotide polymorphisms from paralogous sequence variants and complex polymorphic positions. Sites of shared polymorphism are considered potential sites of gene conversion or double cross-over between the two LCR22s. We found an inverse correlation between regions of paralogous sequence variants that are unique to a given position within one LCR22 and clusters of shared polymorphic sites, suggesting that these clusters depict altered recombination and not remnants of ancestral single nucleotide polymorphisms. We postulate that most shared polymorphic sites are products of past transfers of DNA information between the LCR22s, suggesting that frequent traffic of genetic material may induce genomic instability in the two LCR22s. We also found that gaps up to 1.5 kb long can be transferred between LCR22s. PMID- 16251459 TI - Ascertainment bias in studies of human genome-wide polymorphism. AB - Large-scale SNP genotyping studies rely on an initial assessment of nucleotide variation to identify sites in the DNA sequence that harbor variation among individuals. This "SNP discovery" sample may be quite variable in size and composition, and it has been well established that properties of the SNPs that are found are influenced by the discovery sampling effort. The International HapMap project relied on nearly any piece of information available to identify SNPs-including BAC end sequences, shotgun reads, and differences between public and private sequences-and even made use of chimpanzee data to confirm human sequence differences. In addition, the ascertainment criteria shifted from using only SNPs that had been validated in population samples, to double-hit SNPs, to finally accepting SNPs that were singletons in small discovery samples. In contrast, Perlegen's primary discovery was a resequencing-by-hybridization effort using the 24 people of diverse origin in the Polymorphism Discovery Resource. Here we take these two data sets and contrast two basic summary statistics, heterozygosity and F(ST), as well as the site frequency spectra, for 500-kb windows spanning the genome. The magnitude of disparity between these samples in these measures of variability indicates that population genetic analysis on the raw genotype data is ill advised. Given the knowledge of the discovery samples, we perform an ascertainment correction and show how the post-correction data are more consistent across these studies. However, discrepancies persist, suggesting that the heterogeneity in the SNP discovery process of the HapMap project resulted in a data set resistant to complete ascertainment correction. Ascertainment bias will likely erode the power of tests of association between SNPs and complex disorders, but the effect will likely be small, and perhaps more importantly, it is unlikely that the bias will introduce false-positive inferences. PMID- 16251460 TI - Genetically indistinguishable SNPs and their influence on inferring the location of disease-associated variants. AB - As part of a recent high-density linkage disequilibrium (LD) study of chromosome 20, we obtained genotypes for approximately 30,000 SNPs at a density of 1 SNP/2 kb on four different population samples (47 CEPH founders; 91 UK unrelateds [unrelated white individuals of western European ancestry]; 97 African Americans; 42 East Asians). We observed that approximately 50% of SNPs had at least one genetically indistinguishable partner; i.e., for every individual considered, their genotype at the first locus was identical to their genotype at the second locus, or in LD terms, the SNPs were in "perfect" LD (r2 = 1.0). These "genetically indistinguishable SNPs" (giSNPs) formed into clusters of varying size. The larger the cluster, the greater the tendency to be located within genes and to overlap with giSNP clusters in other population samples. As might be expected for this map density, many giSNPs were located close to one another, thus reflecting local regions of undetected recombination or haplotype blocks. However, approximately 1/3 of giSNP clusters had intermingled, non indistinguishable SNPs with incomplete LD (D' and r2 <1), sometimes spanning hundreds of kilobases, comprising up to 70 indistinguishable markers and overlapping multiple haplotype blocks. These long-range, nonconsecutive giSNPs have implications for disease gene localization by allelic association as evidence for association at one locus will be indistinguishable from that at another locus, even though both loci may be situated far apart. We describe the distribution of giSNPs on this map of chromosome 20 and illustrate the potential impact they can have on association mapping. PMID- 16251461 TI - Genome-wide definitive haplotypes determined using a collection of complete hydatidiform moles. AB - We present genome-wide definitive haplotypes, determined using a collection of 74 Japanese complete hydatidiform moles, each carrying a genome derived from a single sperm. The haplotypes incorporate 281,439 common SNPs, genotyped with a high throughput array-based oligonucleotide hybridization technique. Comparison of haplotypes inferred from pseudoindividuals (constructed from randomized mole pairs) with those of moles showed some switch errors in resolution of phases by the computational inference method. The effects of these errors on local haplotype structure and selection of tag SNPs are discussed. We also show that definitive haplotypes of moles may be useful for elucidation of long-range haplotype structure, and should be more effective for detecting extended haplotype homozygosity indicative of positive selection. PMID- 16251462 TI - Sequence features in regions of weak and strong linkage disequilibrium. AB - We use genotype data generated by the International HapMap Project to dissect the relationship between sequence features and the degree of linkage disequilibrium in the genome. We show that variation in linkage disequilibrium is broadly similar across populations and examine sequence landscape in regions of strong and weak disequilibrium. Linkage disequilibrium is generally low within approximately 15 Mb of the telomeres of each chromosome and noticeably elevated in large, duplicated regions of the genome as well as within approximately 5 Mb of centromeres and other heterochromatic regions. At a broad scale (100-1000 kb resolution), our results show that regions of strong linkage disequilibrium are typically GC poor and have reduced polymorphism. In addition, these regions are enriched for LINE repeats, but have fewer SINE, DNA, and simple repeats than the rest of the genome. At a fine scale, we examine the sequence composition of "hotspots" for the rapid breakdown of linkage disequilibrium and show that they are enriched in SINEs, in simple repeats, and in sequences that are conserved between species. Regions of high and low linkage disequilibrium (the top and bottom quartiles of the genome) have a higher density of genes and coding bases than the rest of the genome. Closer examination of the data shows that whereas some types of genes (including genes involved in immune response and sensory perception) are typically located in regions of low linkage disequilibrium, other genes (including those involved in DNA and RNA metabolism, response to DNA damage, and the cell cycle) are preferentially located in regions of strong linkage disequilibrium. Our results provide a detailed analysis of the relationship between sequence features and linkage disequilibrium and suggest an evolutionary justification for the heterogeneity in linkage disequilibrium in the genome. PMID- 16251463 TI - Segmental duplications and gene conversion: Human luteinizing hormone/chorionic gonadotropin beta gene cluster. AB - Segmental duplicons (>1 kb) of high sequence similarity (>90%) covering >5% of the human genome are characterized by complex sequence variation. Apart from a few well-characterized regions (MHC, beta-globin), the diversity and linkage disequilibrium (LD) patterns of duplicons and the role of gene conversion in shaping them have been poorly studied. To shed light on these issues, we have re sequenced the human Luteinizing Hormone/Chorionic Gonadotropin beta (LHB/CGB) cluster (19q13.32) of three population samples (Estonians, Mandenka, and Han). The LHB/CGB cluster consists of seven duplicated genes critical in human reproduction. In the LHB/CGB region, high sequence diversity, concentration of gene-conversion acceptor sites, and strong LD colocalize with peripheral genes, whereas central loci are characterized by lower variation, gene-conversion donor activity, and breakdown of LD between close markers. The data highlight an important role of gene conversion in spreading polymorphisms among duplicon copies and generating LD around them. The directionality of gene-conversion events seems to be determined by the localization of a predicted recombination "hotspot" and "warm spot" in the vicinity of the most active acceptor genes at the periphery of the cluster. The data suggest that enriched crossover activity in direct and inverted segmental repeats is in accordance with the formation of palindromic secondary structures promoting double-strand breaks rather than fixed DNA sequence motifs. Also, this first detailed coverage of sequence diversity and structure of the LHB/CGB gene cluster will pave the way for studying the identified polymorphisms as well as potential genomic rearrangements in association with an individual's reproductive success. PMID- 16251464 TI - Large-scale recombination rate patterns are conserved among human populations. AB - In humans, most recombination events occur in a small fraction of the genome. These hotspots of recombination show considerable variation in intensity and/or location across species and, potentially, across human populations. On a larger scale, the patterns of recombination rates have been mostly investigated in individuals of European ancestry, and it remains unknown whether the results obtained can be directly applied to other human populations. Here, we investigate this question using genome-wide polymorphism data. We show that population recombination rates recapitulate a large part of the genetic map information, regardless of the population considered. We also show that the ratio of the population recombination rate estimate of two populations is overall constant along the chromosomes. These two observations support the hypothesis that large scale recombination patterns are conserved across human populations. Local deviations from the overall pattern of conservation of the recombination rates can be used to select candidate regions with large polymorphic inversions or under local selection. PMID- 16251465 TI - Genomic regions exhibiting positive selection identified from dense genotype data. AB - The allele frequency spectrum of polymorphisms in DNA sequences can be used to test for signatures of natural selection that depart from the expected frequency spectrum under the neutral theory. We observed a significant (P = 0.001) correlation between the Tajima's D test statistic in full resequencing data and Tajima's D in a dense, genome-wide data set of genotyped polymorphisms for a set of 179 genes. Based on this, we used a sliding window analysis of Tajima's D across the human genome to identify regions putatively subject to strong, recent, selective sweeps. This survey identified seven Contiguous Regions of Tajima's D Reduction (CRTRs) in an African-descent population (AD), 23 in a European-descent population (ED), and 29 in a Chinese-descent population (XD). Only four CRTRs overlapped between populations: three between ED and XD and one between AD and ED. Full resequencing of eight genes within six CRTRs demonstrated frequency spectra inconsistent with neutral expectations for at least one gene within each CRTR. Identification of the functional polymorphism (and/or haplotype) responsible for the selective sweeps within each CRTR may provide interesting insights into the strongest selective pressures experienced by the human genome over recent evolutionary history. PMID- 16251466 TI - Genomic scans for selective sweeps using SNP data. AB - Detecting selective sweeps from genomic SNP data is complicated by the intricate ascertainment schemes used to discover SNPs, and by the confounding influence of the underlying complex demographics and varying mutation and recombination rates. Current methods for detecting selective sweeps have little or no robustness to the demographic assumptions and varying recombination rates, and provide no method for correcting for ascertainment biases. Here, we present several new tests aimed at detecting selective sweeps from genomic SNP data. Using extensive simulations, we show that a new parametric test, based on composite likelihood, has a high power to detect selective sweeps and is surprisingly robust to assumptions regarding recombination rates and demography (i.e., has low Type I error). Our new test also provides estimates of the location of the selective sweep(s) and the magnitude of the selection coefficient. To illustrate the method, we apply our approach to data from the Seattle SNP project and to Chromosome 2 data from the HapMap project. In Chromosome 2, the most extreme signal is found in the lactase gene, which previously has been shown to be undergoing positive selection. Evidence for selective sweeps is also found in many other regions, including genes known to be associated with disease risk such as DPP10 and COL4A3. PMID- 16251467 TI - Calibrating a coalescent simulation of human genome sequence variation. AB - Population genetic models play an important role in human genetic research, connecting empirical observations about sequence variation with hypotheses about underlying historical and biological causes. More specifically, models are used to compare empirical measures of sequence variation, linkage disequilibrium (LD), and selection to expectations under a "null" distribution. In the absence of detailed information about human demographic history, and about variation in mutation and recombination rates, simulations have of necessity used arbitrary models, usually simple ones. With the advent of large empirical data sets, it is now possible to calibrate population genetic models with genome-wide data, permitting for the first time the generation of data that are consistent with empirical data across a wide range of characteristics. We present here the first such calibrated model and show that, while still arbitrary, it successfully generates simulated data (for three populations) that closely resemble empirical data in allele frequency, linkage disequilibrium, and population differentiation. No assertion is made about the accuracy of the proposed historical and recombination model, but its ability to generate realistic data meets a long standing need among geneticists. We anticipate that this model, for which software is publicly available, and others like it will have numerous applications in empirical studies of human genetics. PMID- 16251468 TI - Survey of allelic expression using EST mining. AB - Cis-acting allelic variation in gene regulation is a source of phenotypic variation. Consequently, recent studies have experimentally screened human genes in an attempt to initiate a catalog of genes possessing cis-acting variants. In this study, we use human EST data in dbEST as the source of allelic expression data, and the HapMap database to provide expected allele frequencies in human populations. We demonstrate a greater concordance of allele frequencies estimated from human ESTs in dbEST with those derived from the CEPH HapMap sample representing Caucasians from northern and western Europe, than population samples obtained in Asia and Africa. Deviations between allele frequencies observed in EST databases and the ones obtained from the CEPH HapMap samples may result from common heritable cis-acting variants altering the relative allele distribution in RNA. We provide in silico as well as experimental evidence that this strategy does allow significant enrichment of genes harboring common heritable cis-acting polymorphisms in linkage disequilibrium with expressed alleles. PMID- 16251469 TI - The International HapMap Project Web site. AB - The HapMap Web site at http://www.hapmap.org is the primary portal to genotype data produced as part of the International Haplotype Map Project. In phase I of the project, >1.1 million SNPs were genotyped in 270 individuals from four worldwide populations. The HapMap Web site provides researchers with a number of tools that allow them to analyze the data as well as download data for local analyses. This paper presents step-by-step guides to using those tools, including guides for retrieving genotype and frequency data, picking tag-SNPs for use in association studies, viewing haplotypes graphically, and examining marker-to marker LD patterns. PMID- 16251470 TI - Inference and analysis of haplotypes from combined genotyping studies deposited in dbSNP. AB - In the attempt to understand human variation and the genetic basis of complex disease, a tremendous number of single nucleotide polymorphisms (SNPs) have been discovered and deposited into NCBI's dbSNP public database. More than 2.7 million SNPs in the database have genotype information. This data provides an invaluable resource for understanding the structure of human variation and the design of genetic association studies. The genotypes deposited to dbSNP are unphased, and thus, the haplotype information is unknown. We applied the phasing method HAP to obtain the haplotype information, block partitions, and tag SNPs for all publicly available genotype data and deposited this information into the dbSNP database. We also deposited the orthologous chimpanzee reference sequence for each predicted haplotype block computed using the UCSC BLASTZ alignments of human and chimpanzee. Using dbSNP, researchers can now easily perform analyses using multiple genotype data sets from the same genomic regions. Dense and sparse genotype data sets from the same region were combined to show that the number of common haplotypes is significantly underestimated in whole genome data sets, while the predicted haplotypes over the common SNPs are consistent between studies. To validate the accuracy of the predictions, we bench-marked HAP's running time and phasing accuracy against PHASE. Although HAP is slightly less accurate than PHASE, HAP is over 1000 times faster than PHASE, making it suitable for application to the entire set of genotypes in dbSNP. PMID- 16251471 TI - Decline of contractility during ischemia-reperfusion injury: actin glutathionylation and its effect on allosteric interaction with tropomyosin. AB - The severity and duration of ischemia-reperfusion injury is hypothesized to play an important role in the ability of the heart subsequently to recover contractility. Permeabilized trabeculae were prepared from a rat model of ischemia-reperfusion injury to examine the impact on force generation. Compared with the control perfused condition, the maximum force (F(max)) per cross sectional area and the rate of tension redevelopment of Ca(2+)-activated trabeculae fell by 71% and 44%, respectively, during ischemia despite the availability of a high concentration of ATP. The reduction in F(max) with ischemia was accompanied by a decline in fiber stiffness, implying a drop in the absolute number of attached cross bridges. However, the declines during ischemia were largely recovered after reperfusion, leading to the hypothesis that intrinsic, reversible posttranslational modifications to proteins of the contractile filaments occur during ischemia-reperfusion injury. Examination of thin-filament proteins from ischemic or ischemia-reperfused hearts did not reveal proteolysis of troponin I or T. However, actin was found to be glutathionylated with ischemia. Light-scattering experiments demonstrated that glutathionylated G actin did not polymerize as efficiently as native G-actin. Although tropomyosin accelerated the time course of native and glutathionylated G-actin polymerization, the polymerization of glutathionylated G-actin still lagged native G-actin at all concentrations of tropomyosin tested. Furthermore, cosedimentation experiments demonstrated that tropomyosin bound glutathionylated F-actin with significantly reduced cooperativity. Therefore, glutathionylated actin may be a novel contributor to the diverse set of posttranslational modifications that define the function of the contractile filaments during ischemia-reperfusion injury. PMID- 16251472 TI - Bipolar assembly of caveolae in retinal pigment epithelium. AB - Caveolae and their associated structural proteins, the caveolins, are specialized plasmalemmal microdomains involved in endocytosis and compartmentalization of cell signaling. We examined the expression and distribution of caveolae and caveolins in retinal pigment epithelium (RPE), which plays key roles in retinal support, visual cycle, and acts as the main barrier between blood and retina. Electron microscopic observation of rat RPE, in situ primary cultures of rat and human RPE and a rat RPE cell line (RPE-J) demonstrated in all cases the presence of caveolae in both apical and basolateral domains of the plasma membrane. Caveolae were rare in RPE in situ but were frequent in primary RPE cultures and in RPE-J cells, which correlated with increased levels in the expression of caveolin-1 and -2. The bipolar distribution of caveolae in RPE is striking, as all other epithelial cells examined to date (liver, kidney, thyroid, and intestinal) assemble caveolae only at the basolateral side. This might be related to the nonpolar distribution of both caveolin-1 and 2 in RPE because caveolin-2 is basolateral and caveolin-1 nonpolar in other epithelial cells. The bipolar localization of plasmalemmal caveolae in RPE cells may reflect specialized roles in signaling and trafficking important for visual function. PMID- 16251473 TI - Type II skeletal myofibers possess unique properties that potentiate mitochondrial H(2)O(2) generation. AB - Mitochondrial dysfunction is implicated in a number of skeletal muscle pathologies, most notably aging-induced atrophy and loss of type II myofibers. Although oxygen-derived free radicals are thought to be a primary cause of mitochondrial dysfunction, the underlying factors governing mitochondrial superoxide production in different skeletal myofiber types is unknown. Using a novel in situ approach to measure H(2)O(2) production (indicator of superoxide formation) in permeabilized rat skeletal muscle fiber bundles, we found that mitochondrial free radical leak (H(2)O(2) produced/O(2) consumed) is two- to threefold higher (P < 0.05) in white (WG, primarily type IIB fibers) than in red (RG, type IIA) gastrocnemius or soleus (type I) myofibers during basal respiration supported by complex I (pyruvate + malate) or complex II (succinate) substrates. In the presence of respiratory inhibitors, maximal rates of superoxide produced at both complex I and complex III are markedly higher in RG and WG than in soleus muscle despite approximately 50% less mitochondrial content in WG myofibers. Duplicate experiments conducted with +/-exogenous superoxide dismutase revealed striking differences in the topology and/or dismutation of superoxide in WG vs. soleus and RG muscle. When normalized for mitochondrial content, overall H(2)O(2) scavenging capacity is lower in RG and WG fibers, whereas glutathione peroxidase activity, which is largely responsible for H(2)O(2) removal in mitochondria, is similar in all three muscle types. These findings suggest that type II myofibers, particularly type IIB, possess unique properties that potentiate mitochondrial superoxide production and/or release, providing a potential mechanism for the heterogeneous development of mitochondrial dysfunction in skeletal muscle. PMID- 16251474 TI - Nongenomic regulation by aldosterone of the epithelial NHE3 Na(+)/H(+) exchanger. AB - The relevance of nongenomic pathways to regulation of epithelial function by aldosterone is poorly understood. Recently, we demonstrated that aldosterone inhibits transepithelial HCO(3)(-) absorption in the renal medullary thick ascending limb (MTAL) through a nongenomic pathway. Here, we examined the transport mechanism(s) responsible for this regulation, focusing on Na(+)/H(+) exchangers (NHE). In the MTAL, apical NHE3 mediates H(+) secretion necessary for HCO(3)(-) absorption; basolateral NHE1 influences HCO(3)(-) absorption by regulating apical NHE3 activity. In microperfused rat MTALs, the addition of 1 nM aldosterone rapidly decreased HCO(3)(-) absorption by 30%. This inhibition was unaffected by three maneuvers that inhibit basolateral Na(+)/H(+) exchange and was preserved in MTALs from NHE1 knockout mice, ruling out the involvement of NHE1. In contrast, exposure to aldosterone for 15 min caused a 30% decrease in apical Na(+)/H(+) exchange activity over the intracellular pH range from 6.5 to 7.7, due to a decrease in V(max). Inhibition of HCO(3)(-) absorption by aldosterone was not affected by 0.1 mM lumen Zn(2+) or 1 mM lumen DIDS, arguing against the involvement of an apical H(+) conductance or apical K(+)-HCO(3)(-) cotransport. These results demonstrate that aldosterone inhibits HCO(3)(-) absorption in the MTAL through inhibition of apical NHE3, and identify NHE3 as a target for nongenomic regulation by aldosterone. Aldosterone may influence a broad range of epithelial transport functions important for extracellular fluid volume and acid-base homeostasis through direct regulation of this exchanger. PMID- 16251475 TI - Regulation of homocysteine-induced MMP-9 by ERK1/2 pathway. AB - Homocysteine (Hcy) induces matrix metalloproteinase (MMP)-9 in microvascular endothelial cells (MVECs). We hypothesized that the ERK1/2 signaling pathway is involved in Hcy-mediated MMP-9 expression. In cultured MVECs, Hcy induced activation of ERK, which was blocked by PD-98059 and U0126 (MEK inhibitors). Pretreatment with BAPTA-AM, staurosporine (PKC inhibitor), or Go6976 (specific inhibitor for Ca(2+)-dependent PKC) abrogated ERK phosphorylation, suggesting the role of Ca(2+) and Ca(2+)-dependent PKC in Hcy-induced ERK activation. ERK phosphorylation was suppressed by pertussis toxin (PTX), suggesting the involvement of G protein-coupled receptors (GPCRs) in initiating signal transduction by Hcy and leading to ERK activation. Pretreatment of MVECs with genistein, BAPTA-AM, or thapsigargin abrogated Hcy-induced ERK activation, suggesting the involvement of the PTK pathway in Hcy-induced ERK activation, which was mediated by intracellular Ca(2+) pool depletion. ERK activation was attenuated by preincubation with N-acetylcysteine (NAC) and SOD, suggesting the role of oxidation in Hcy-induced ERK activation. Pretreatment with an ERK1/2 blocker (PD-98059), staurosporine, folate, or NAC modulated Hcy-induced MMP-9 activation as measured using zymography. Our results provide evidence that Hcy triggers the PTX-sensitive ERK1/2 signaling pathway, which is involved in the regulation of MMP-9 in MVECs. PMID- 16251476 TI - ERK/MAPK regulates the Kv4.2 potassium channel by direct phosphorylation of the pore-forming subunit. AB - Kv4.2 is the primary pore-forming subunit encoding A-type currents in many neurons throughout the nervous system, and it also contributes to the transient outward currents of cardiac myocytes. A-type currents in the dendrites of hippocampal CA1 pyramidal neurons are regulated by activation of ERK/MAPK, and Kv4.2 is the likely pore-forming subunit of that current. We showed previously that Kv4.2 is directly phosphorylated at three sites by ERK/MAPK (T602, T607, and S616). In this study we determined whether direct phosphorylation of Kv4.2 by ERK/MAPK is responsible for the regulation of the A-type current observed in neurons. We made site-directed mutants, changing the phosphosite serine (S) or threonine (T) to aspartate (D) to mimic phosphorylation. We found that the T607D mutation mimicked the electrophysiological changes elicited by ERK/MAPK activation in neurons: a rightward shift of the activation curve and an overall reduction in current compared with wild type (WT). Surprisingly, the S616D mutation caused the opposite effect, a leftward shift in the activation voltage. K(+) channel-interacting protein (KChIP)3 ancillary subunit coexpression with Kv4.2 was necessary for the T607D effect, as the T607D mutant when expressed in the absence of KChIP3 was not different from WT Kv4.2. These data suggest that direct phosphorylation of Kv4.2 at T607 is involved in the dynamic regulation of the channel function by ERK/MAPK and an interaction of the primary subunit with KChIP is also necessary for this effect. Overall these studies provide new insights into the structure-function relationships for MAPK regulation of membrane ion channels. PMID- 16251477 TI - Multiple eicosanoid-activated nonselective cation channels regulate B-lymphocyte adhesion to integrin ligands. AB - Arachidonic acid (AA) is a substrate for a variety of proinflammatory mediators, which are generated by cyclooxygenases (COXs), lipoxygenases (LOXs), and cytochrome P-450 (CYP450) enzymes. COX (e.g., PGs and prostacyclins) and LOX (e.g., leukotrienes) products have well-established proinflammatory roles; however, little is known about the functions of CYP450 products in leukocytes. We previously found that mechanical strain generated by subjecting lymphocytes to hypotonic challenge triggered AA production and that two CYP450 products of AA, 5,6-epoxyeicosatrienoic acid (5,6-EET) and 20-hydroxyeicosatetraenoic acid (20 HETE), as well as a product of LOX, 5-(S)-hydroperoxyeicosatetrenoic acid (5 HPETE), induced Ca(2+) entry into primary B cells. The main goal of the present studies, therefore, was to define the biophysically properties of eicosanoid activated channels responsible for Ca(2+) entry and the physiological consequences of activating these channels, including their role in mechanical signaling. We found that 5,6-EET, 20-HETE, and 5-HPETE each activated distinct Ca(2+)-permeant nonselective cation channels (NSCCs) in primary B cells. These NSCCs each regulate plasma membrane potential and B-cell adhesion to integrin ligands ICAM-1 and VCAM-1. Thus our data demonstrate that proinflammatory mediators produced in response to osmotic and/or physical stress play a direct role in regulating the B-cell membrane potential and their adhesion to specific ECM proteins. These results not only have important implications for understanding normal mechanisms of B-cell activation, differentiation, and trafficking but also point to novel targets for modulating the pathogenesis of B cell-mediated inflammatory diseases. PMID- 16251478 TI - Innate immune responses of human tracheal epithelium to Pseudomonas aeruginosa flagellin, TNF-alpha, and IL-1beta. AB - We measured innate immune responses by primary human tracheal epithelial (HTE) cells grown as confluent, pseudostratified layers during exposure to inflammatory activators on apical vs. basolateral surfaces. Apical Pseudomonas aeruginosa strain PAK (but not flagellin mutant PAK.fliC), flagellin, and flagellin + PAK.fliC activated NF-kappaB and IL-8 expression and secretion. In contrast, HTE cells were insensitive to LPS compared to flagellin. Flagellin activated NF kappaB in columnar but not basal cells. IL-1beta + TNF-alpha elicited responses similar to those of flagellin. Basolateral flagellin or IL-1beta + TNF-alpha caused 1.5- to 4-fold larger responses, consistent with the fact that NF-kappaB activation occurred in both columnar and basal cells. MyD88 (toll receptor associated adapter), IL-1 receptor (IL1R)1, and TNF-alpha receptor (TNFR)1 were expressed in columnar and basal cells. ZO-1 was localized to tight junctions of columnar cells but not to basal cells. We infer the following. 1) Flagellin is necessary and sufficient to trigger inflammatory responses in columnar cells during accumulation of P. aeruginosa in the airway surface liquid (ASL); columnar cells express toll-like receptor 5 and MyD88, often associated with flagellin activated cell signaling. 2) IL-1beta + TNF-alpha in the ASL also activate columnar cells, and these cells also express IL1R1 and TNFR1. 3) Apical flagellin, IL-1beta, and TNF-alpha do not activate basal cells because tight junctions between columnar cells prevent access from the apical surface to the basal cells. 4) Exposure of basolateral surfaces to inflammatory activators elicits larger responses because both columnar and basal cells are activated, likely because both cell types express receptors for flagellin, IL-1beta, and TNF alpha. PMID- 16251479 TI - Psychological distress and the impact of social support on fathers and mothers of pediatric cancer patients: long-term prospective results. AB - OBJECTIVE: To explore the effects of social support on psychological distress of parents of pediatric cancer patients using a prospective design over a 5-year period. METHODS: Parents of children diagnosed with cancer participated at diagnosis (T1), 6 months (T2), 12 months (T3), and 5 years later (T4). Instruments The General Health Questionnaire and the Social Support List (SSL) measuring amount of support, (dis)satisfaction with support, and negative interactions were administered. RESULTS: Psychological distress and amount of support received decreased significantly from diagnosis to T4. No significant change in (dis)satisfaction with support and negative interactions was found. Social support variables did not show any concurrent or prospective significant effect on mothers' distress at T4. Dissatisfaction with support showed a significant unique concurrent effect on fathers' distress at T4 and negative interactions had a prospective unique effect. CONCLUSIONS: Dissatisfaction with support and negative interactions that fathers experienced significantly affected their levels of psychological distress. No such effect was found for mothers. PMID- 16251480 TI - Commentary: a look at ourselves in the mirror. PMID- 16251481 TI - Cell cycle inhibition by sodium arsenite in primary embryonic rat midbrain neuroepithelial cells. AB - Arsenite (As3+) exposure during development has been associated with neural tube defects and other structural malformations, and with behavioral alterations including altered locomotor activity and operant learning. The molecular mechanisms underlying these effects are uncertain. Because arsenic can cross the placenta and accumulate in the developing neuroepithelium, we examined cell cycling effects of sodium arsenite (As3+ 0, 0.5, 1, 2, and 4 microM) on embryonic primary rat midbrain (gestational day [GD] 12) neuroepithelial cells over 48 h. There was a concentration- and time-dependent As3+-induced reduction in cell viability assessed by neutral red dye uptake assay but minimal apoptosis at concentrations below 4 microM. Morphologically, apoptosis was not apparent until 4 microM at 24 h, which was demonstrated by a marginal but statistically significant increase in cleaved caspase-3/7 activity. Cell cycling effects over several rounds of replication were determined by continuous 5-bromo-2' deoxyuridine (BrdU) labeling and bivariate flow cytometric Hoechst-Propidium Iodide analysis. We observed a time- and concentration-dependent inhibition of cell cycle progression as early as 12 h after exposure (> or =0.5 microM). In addition, data demonstrated a concentration-dependent increase in cytostasis within all cell cycle phases, a decreased proportion of cells able to reach the second cell cycle, and a reduced cell cycle entry from gap 1 phase (G1). The proportion of affected cells and the severity of the cell cycle perturbation, which ranged from a decreased transition probability to complete cytostasis in all cell cycle phases, were also found to be concentration-dependent. Together, these data support a role for perturbed cell cycle progression in As3+ mediated neurodevelopmental toxicity. PMID- 16251482 TI - Cisplatin-induced hepatotoxicity is enhanced by elevated expression of cytochrome P450 2E1. AB - In this study, the possible potentiation of cisplatin-induced hepatotoxicity by cytochrome P450 2E1 (CYP2E1) was examined both in vitro and in vivo. Transfected HepG2 cells expressing CYP2E1 (E47 cells) and not expressing CYP2E1 (C34 cells) were used as an in vitro model, and mice drinking 2% acetone for 7 days to induce CYP2E1 were used as an in vivo model. Exposure of E47 cells to cisplatin caused a much greater loss of cell viability, more striking depletion of reduced glutathione (GSH), and higher reactive oxygen species (ROS) production as compared with C34 cells. The prooxidant L-buthionine-[R,S]-sulfoximine (BSO), which depletes GSH, enhanced cisplatin-induced loss of cell viability, whereas the antioxidant glutathione ethyl ester, or the iron chelator deferoxamine mesylate (DFO) protected against the cisplatin-induced loss of E47 cell viability. Diallyl sulfide (DAS), an inhibitor of CYP2E1, also protected against the cisplatin toxicity in the E47 cells. After being injected with cisplatin (ip, 45 mg/kg), mice drinking 2% acetone with increased CYP2E1 levels exhibited elevated levels of serum ALT and AST, liver caspase-3 activity and positive staining of TUNEL increased, and histopathology indicated the presence of necrotic foci in livers of acetone plus cisplatin-treated mice. Lipid peroxidation and protein oxidation as indicated by carbonyl formation, staining of 3-nitrotyrosine (3-NT) and iron were higher in the cisplatin plus acetone group, compared with cisplatin alone group. Both in vitro and in vivo results indicate that elevated CYP2E1 enhances cisplatin-induced hepatotoxicity, and the mechanism may involve increased production of ROS and oxidative stress. PMID- 16251483 TI - DNA hypermethylation of promoter of gene p53 and p16 in arsenic-exposed people with and without malignancy. AB - Chronic arsenic exposure is known to produce arsenicosis and cancer. To ascertain whether perturbation of methylation plays a role in such carcinogenesis, the degree of methylation of p53 and p16 gene in DNA obtained from blood samples of people chronically exposed to arsenic and skin cancer subjects was studied. Methylation-specific restriction endonuclease digestion followed by polymerase chain reaction (PCR) of gene p53 and bisulfite treatment followed by methylation sensitive PCR of gene p16 have been carried out to analyze the methylation status of the samples studied. Significant DNA hypermethylation of promoter region of p53 gene was observed in DNA of arsenic-exposed people compared to control subjects. This hypermethylation showed a dose-response relationship. Further, hypermethylation of p53 gene was also observed in arsenic-induced skin cancer patients compared to subjects having skin cancer unrelated to arsenic, though not at significant level. However, a small subgroup of cases showed hypomethylation with high arsenic exposure. Significant hypermethylation of gene p16 was also observed in cases of arsenicosis exposed to high level of arsenic. In man, arsenic has the ability to alter DNA methylation patterns in gene p53 and p16, which are important in carcinogenesis. PMID- 16251484 TI - Inorganic mercury inhibits the activation of LAT in T-cell receptor-mediated signal transduction. AB - Little is known as to the molecular mechanisms involved with mercury intoxication at very low levels. Although the mechanism is not known, animal studies have nevertheless shown that low levels of mercury may target the immune system. Inorganic mercury (Hg2+) at very low (but non-toxic) levels can disrupt immune system homeostasis, in that genetically susceptible rodents develop idiosyncratic autoimmune disease, which is associated with defective T-cell function. T lymphocyte function is intimately coupled to the T-cell receptor. We have previously reported that on a molecular level, low concentrations of Hg2+ disrupt signaling from the T-cell receptor by interfering with activation of Ras and ERK MAP kinase. In this report we expand upon those results by showing that in T lymphocytes exposed to low concentration of Hg2+, Ras fails to become properly activated because upstream of Ras in the T cell signal transduction pathway, the important scaffolding element Linker for Activation of T Cells (LAT) fails to become properly phosphorylated. Hypo-phosphorylation of LAT occurs, because upstream of LAT, the LAT reactive tyrosine kinase ZAP-70 is also not properly activated in Hg2+ treated cells. PMID- 16251485 TI - A toxicogenomics approach to identify new plausible epigenetic mechanisms of ochratoxin a carcinogenicity in rat. AB - Ochratoxin A (OTA) is a mycotoxin occurring naturally in a wide range of food commodities. In animals, it has been shown to cause a variety of adverse effects, nephrocarcinogenicity being the most prominent. Because of its high toxic potency and the continuous exposure of the human population, OTA has raised public health concerns. There is significant debate on how to use the rat carcinogenicity data to assess the potential risk to humans. In this context, the question of the mechanism of action of OTA appears of key importance and was studied through the application of a toxicogenomics approach. Male Fischer rats were fed OTA for up to 2 years. Renal tumors were discovered during the last 6 months of the study. The total tumor incidence reached 25% at the end of the study. Gene expression profile was analyzed in groups of animals taken in intervals from 7 days to 12 months. Tissue-specific responses were observed in kidney versus liver. For selected genes, microarray data were confirmed at both mRNA and protein levels. In kidney, several genes known as markers of kidney injury and cell regeneration were significantly modulated by OTA. The expression of genes known to be involved in DNA synthesis and repair, or genes induced as a result of DNA damage, was only marginally modulated. Very little or no effect was found amongst genes associated with apoptosis. Alterations of gene expression indicating effects on calcium homeostasis and a disruption of pathways regulated by the transcription factors hepatocyte nuclear factor 4 alpha (HNF4alpha) and nuclear factor-erythroid 2 related factor 2 (Nrf2) were observed in the kidney but not in the liver. Previous data have suggested that a reduction in HNF4alpha may be associated with nephrocarcinogenicity. Many Nrf2-regulated genes are involved in chemical detoxication and antioxidant defense. The depletion of these genes is likely to impair the defense potential of the cells, resulting in chronic elevation of oxidative stress in the kidney. The inhibition of defense mechanism appears as a highly plausible new mechanism, which could contribute to OTA carcinogenicity. PMID- 16251486 TI - Editorial: developmental and situational transitions. PMID- 16251487 TI - Editorial: aesthetics, art, and holistic nursing. PMID- 16251488 TI - Health care choices: complementary therapy, chronic illness, and older rural dwellers. AB - PURPOSE: To better understand elderly rural dwellers use of complementary and alternative therapies (CAM), perception of efficacy, and their sources of information. METHOD: Ten participants between ages 60 and 80 years were interviewed by telephone. Qualitative analysis for content and themes was done. FINDINGS: Self-directed practices such as vitamin, mineral, and herbal supplements, taken to compensate for perceived dietary deficiencies, were cited most frequently. Therapies used included biofeedback, massage, chiropractic, and podiatry. Information was obtained from the physician or nurse practitioner and supplemented with material from other sources. CONCLUSIONS: Respondents were discriminating users of the products they selected and were satisfied with the results, although they did not always have accurate understanding of the effects of the supplement or therapy. IMPLICATIONS: Health professionals need to have knowledge about CAM and to include questions about use during patient encounters. Providers must take time to answer questions and provide user friendly information. PMID- 16251489 TI - Efficacy of frequent mantram repetition on stress, quality of life, and spiritual well-being in veterans: a pilot study. AB - PURPOSE: Silent, frequent repetition of a mantram-a word or phrase with spiritual significance, sometimes called a Holy Name-is an ancient form of prayer that may reduce stress and related symptoms. The authors tested the feasibility and efficacy of a 5-week (90-min per week) intervention on mantram repetition in a sample of ambulatory veterans. METHOD: Aconvenience sample (N= 62) of outpatient veterans participated in the study by completing pre-and posttest self-report questionnaires on stress, anxiety, anger, quality of life, and spiritual well being. Wrist-worn counters were provided to track mantram practice. FINDINGS: Mantram repetition significantly reduced symptoms of stress and anxiety and improved quality of life and spiritual well-being. CONCLUSION: Additional research using a larger sample size and control group is needed to further substantiate the benefits of this intervention. IMPLICATIONS: Frequent, silent mantram repetition is easily taught and could be used by nurses and patients for managing stress and increasing well-being. PMID- 16251490 TI - Utility of nutraceutical products marketed for cognitive and memory enhancement. AB - This article identifies a convenience sample of 14 memory-enhancing herbal products that were found to be available commercially, examines their active ingredients, states their claims, and evaluates the available evidence to determine their efficacy. The analyses identified four problematic areas. First, a majority of the products use cognitive terminology, which leads consumers to anticipate an intended cognitive benefit. Second, some ingredients are completely homeopathic and contain components not known outside of the homeopathic field. Third, the evidence of treatment efficacy is often contradictory, because products are recommended for purposes other than cognitive or memory loss. Finally, the manufacturers of the product have usually conducted the research on individual products. Until more research is available, it is suggested that holistic nursing professionals exercise caution in recommending nutraceuticals to their patients/clients for the use of cognitive improvement or memory enhancement. PMID- 16251491 TI - A journey in holistic growth. AB - New ways of learning are necessary for novices who desire to mature in holistic knowledge and educational methods. The purpose of this article is to share an experience in holistic growth, gained through an academic exercise. Students in a nontraditional nursing course titled "Women, Witches, and Healing" were asked to describe "What it means to be a woman (or man) in your world." The exercise, done spontaneously at the end of an all-day seminar, yielded impressive responses. Using qualitative methods, the responses were compiled into single writings. Because of the small number of men (4) only the writings from the women were useful. The writings from the women (26) were revised several times before acceptable by all. The final composition is a tribute not only to those women but also to all of us who desire to be more than our usual roles suggest. PMID- 16251492 TI - My life as a holistic nurse. AB - This article explores a nurse's personal experience with holistic nursing. The author describes aspects of her physical, emotional, and spiritual transformation to foster her own self-care as well as her use of human-to-human interactions and the holistic caring process to create a healing environment for her clients. PMID- 16251493 TI - Midwives at the exit. PMID- 16251495 TI - Low oxygen concentrations inhibit trophoblast cell invasion from early gestation placental explants via alterations in levels of the urokinase plasminogen activator system. AB - Extravillous trophoblast cell (EVT) invasion in early pregnancy occurs in a relatively low-oxygen environment. The role of oxygen in regulation of EVT invasion remains controversial. We hypothesized that 1) culture in 3% oxygen inhibits EVT invasion compared with culture at 8% or 20% oxygen and 2) inhibition of invasion is due to alterations in levels of components of the urokinase plasminogen activator (PLAU, uPA) system rather than through increased apoptosis and/or decreased proliferation. Placental samples (8-10, 12-14, and 16-20 wk gestation) were obtained from women undergoing elective surgical termination of pregnancy or after cesarean section delivery (term) at the Royal Victoria Infirmary, Newcastle upon Tyne, U.K. EVT invasion from placental explants cultured at 3%, 8%, or 20% oxygen was assessed using Matrigel invasion assays. Invasion was assessed on Day 6, explants were harvested for analysis of apoptosis and proliferation, and medium was stored for analysis of PLAU system components by ELISA and casein zymography. Culture at 3% oxygen inhibited EVT invasion. PLAU receptor and plasminogen activator inhibitor-2 protein levels were increased and PLAU activity decreased in these cultures. There was no difference in the proliferation in explants cultured at the three different oxygen concentrations. Apoptosis, assessed by M30 immunostaining, was increased in EVT at both 3% and 8% oxygen. The reduction in the invasive capacity of EVT cultured at 3% oxygen appears to be mediated both by a general inhibition of the PLAU system and a decrease in the number of cells available to invade. PMID- 16251496 TI - Serine peptidase HTRA3 is closely associated with human placental development and is elevated in pregnancy serum. AB - HTRA3 is a newly identified serine peptidase of the mammalian HTRA (high temperature requirement factor A) family, that is upregulated dramatically during mouse placental development. The current study determined whether HTRA3 was involved in human placentation. During the menstrual cycle, HTRA3 was expressed primarily in the endometrial glands, being significantly upregulated toward the mid- to late secretory phases; prominent expression in the stroma detected only in the decidual cells in the late secretory phase. Thus, overall endometrial HTRA3 expression was highest in the late secretory phase, when the endometrium is prepared for maternal-trophoblast interaction. During the first trimester of pregnancy, both glandular and decidual HTRA3 expression increased further with the decidual upregulation being highly significant. The strong link between HTRA3 expression and endometrial stromal cell decidualization was further established in an in vitro model using primary endometrial stromal cells. HTRA3 was also expressed by certain trophoblast subtypes in the first-trimester placenta: strongly in the villous syncytiotrophoblast, trophoblast shell, and endovascular trophoblast and weakly in the distal portion of the trophoblast cell columns but not in villous cytotrophoblast, the proximal region of the cell columns, or interstitial trophoblast. Upregulation of HTRA3 expression in association with placental development was revealed by a significant elevation of this protein in the maternal serum during the first trimester. We thus propose that HTRA3 is a previously unrecognized factor closely associated with and potentially important for human placentation. This study established crucial groundwork for future investigations toward establishing the physiological roles of HTRA3 in human placentation. PMID- 16251497 TI - An epididymal form of cauxin, a carboxylesterase-like enzyme, is present and active in mammalian male reproductive fluids. AB - Mass spectrometric analysis of a prion protein (PrP)-containing complex isolated from ram cauda epididymal fluid revealed a protein that showed homology to a carboxylesterase-like protein previously identified in cat urine (cauxin). Using anti-cauxin antibodies, immunoreactive bands were detected in corpus and cauda epididymal fluid from all mammals tested (ram, boar, mouse, and cat). In the ram, the protein was also present in seminal fluid but not found to be associated with sperm. The bands reacting with the anti-cauxin antibody coincided with those having esterase activity in a zymographic assay and its levels paralleled the esterase activity of native epididymal fluids. A partial nucleotide sequence of 1143 bp, corresponding to 380 amino acids, was obtained by RT-PCR amplification from total RNA from the corpus epididymis (zone 6). The deduced protein sequence shows a high degree of homology (up to 90%) with the different cauxin proteins found in databases but only up to 60% with other known carboxylesterases. By PCR, strong mRNA expression was found in the corpus and cauda epididymis, while the testis, kidney, and caput epididymis had low expression. No mRNA was detected in the lung, heart, or liver. These data demonstrate that an epididymal form of the cauxin enzyme is secreted into mammalian epididymal fluid. In the ram, it is associated with a high molecular-weight PrP-associated complex and may be responsible for the majority of the esterase activity in the cauda epididymal fluid of this species. PMID- 16251499 TI - Trophoblast CD274 (B7-H1) is differentially expressed across gestation: influence of oxygen concentration. AB - Modulation of the maternal immune system by the placenta is a mechanism by which the fetus ensures its own survival in a genetically foreign environment. The immunoinhibitor CD274 (also called B7-H1 or PD-L1) is highly expressed in the placenta, positioned to interact with maternal leukocytes. Further, immunoblot analysis of first- and second-trimester placental lysates showed that CD274 expression is low in the first trimester but dramatically rises around the onset of the second trimester. As this coincides with the expected onset of maternal blood flow to the placenta and a corresponding rise in local oxygen tension, we explored the possibility that oxygen regulates CD274 expression in trophoblast cells by culturing term trophoblast cells under oxygen concentrations similar to those found in vivo. Indeed, CD274 protein levels paralleled the in vivo situation: expression increased with rising oxygen concentrations. Furthermore, downregulation of CD274 mRNA by low oxygen was rapid, occurring within 4-12 h. We conclude that oxygen is a potential mediator of CD274 expression in vivo such that it is induced coincidentally on exposure of fetal tissues to maternal blood. Further, the regulation of this immunomodulator by oxygen may implicate its alteration during and involvement in the pathogenesis of complications of pregnancy such as preeclampsia. PMID- 16251500 TI - Equine sperm membrane phase behavior: the effects of lipid-based cryoprotectants. AB - The plasma membrane of sperm can undergo lipid phase separation during freezing, resulting in irreversible damage to the cell. The objective of our study was to examine the membrane phase behavior of equine spermatozoa in the absence and presence of lipid-based cryoprotectants. Biophysical properties of sperm membranes were investigated with Fourier-transform infrared spectroscopy. Compared to fresh untreated sperm, postthaw untreated sperm showed extensive lipid phase separation and rearrangement. In contrast, postthaw sperm that were cryopreserved in egg phosphatidylcholine (egg PC)- or soy phosphatidylcholine (soy PC)-based diluents showed similar lipid phase behavior to that of fresh, untreated sperm. Studies with a deuterium-labeled PC lipid (POPCd-31) suggest that exogenous lipid from the diluents are strongly associated with the sperm membrane, and scanning electron microscopy images of treated sperm show the presence of lipid aggregates on the membrane surface. Thus, the exogenous lipid does not appear to be integrated into the sperm membrane after cryopreservation. When compared to a standard egg-yolk-based diluent (INRA 82), the soy and egg PC media preserved viability and motility equally well in postthaw sperm. A preliminary fertility study determined that sperm cryopreserved in the soy PC based medium were capable of fertilization at the same rate as sperm frozen in the conventional INRA 82 medium. Our results show that pure lipid-based diluents can prevent membrane damage during cryopreservation and perform as well as a standard egg-yolk-based diluent in preserving sperm viability, motility, and fertility. PMID- 16251501 TI - Age-associated changes in mouse oocytes during postovulatory in vitro culture: possible role for meiotic kinases and survival factor BCL2. AB - To elucidate molecular mechanisms underlying oocyte senescence, we investigated whether oocytes from female mice of advanced reproductive age exhibit a precocious postovulatory aging that, in turn, may be responsible for the precocious activation of an apoptotic program. During a 9-h in vitro culture, the frequency of oocytes showing MII aberrations, spontaneous activation, and cellular fragmentation increased in old oocytes (P < 0.05), whereas it did not change in the young group. In old oocytes, the activities of MPF (a complex of the cyclin-dependent kinase cdc2 and cyclin B1) and MAPK (mitogen-activated protein kinase) decreased precociously, showing a first drop as early as 3 h after the beginning of in vitro culture (P < 0.05). Immunoblotting and immunocytochemical analysis revealed that, in oocytes of the old group, reduction of BCL2 expression at protein level occurred earlier than in the young group (P < 0.05) and was not associated to the loss of BCL2 transcripts detected by RT-PCR. These changes are followed by an abrupt increase of the rate of TUNEL-positive oocytes after 24 h of culture to a value of 67% +/- 6%. Exposure of young oocytes to 20 microM roscovitine or 20 microM U0126, specific inhibitors of MPF and MAPK, resulted in the decreased percentage of oocytes showing positive immunostaining for BCL2 and in an increased rate of DNA fragmentation. Present results suggest that the developmental competence of oocytes ovulated by aging mice may be negatively influenced by a downregulation of MPF and MAPK activities that in turn induces the activation of a proapoptotic signaling pathway. PMID- 16251503 TI - High-frequency stimulation together with adrenoceptor activation facilitates the maintenance of long-term potentiation at visual cortical inhibitory synapses. AB - Long-term potentiation (LTP) at inhibitory synapses of rat visual cortex requires firing of presynaptic cells for maintenance, at least at a low frequency. We examined the roles of adrenoceptors in this LTP maintenance. Although high frequency stimulation (HFS) failed to produce LTP in normal Ca2+ medium, it produced pathway-specific LTP with addition of noradrenaline to the medium soon after HFS. However, this LTP disappeared after washout of noradrenaline. HFS applied during noradrenaline application produced LTP persisting even after washout, indicating that HFS together with adrenoceptor activation makes the adrenergic facilitation enduring. After washout, LTP was produced further by HFS of the conditioned, but not the unconditioned, pathway by the first HFS. Pharmacological examination demonstrated that alpha2 and beta, but not alpha1, receptors facilitated LTP maintenance synergistically. Bath application, but not postsynaptic loading, of either the adenylyl cyclase activator forskolin or the protein kinase C (PKC) activator phorbol ester facilitated LTP maintenance. These results suggest that adrenergic facilitation of LTP maintenance is mediated by presynaptic adrenoceptors via a subfamily of adenylyl cyclases stimulated by Gsalpha, Gibetagamma, and PKC. Thus, it is likely that the activity of noradrenergic afferents takes part in the control of LTP duration at visual cortical inhibitory synapses. PMID- 16251502 TI - Exogenous nitric oxide stimulates cell proliferation via activation of a mitogen activated protein kinase pathway in ovine fetoplacental artery endothelial cells. AB - Sodium nitroprusside (SNP), a nitric oxide (NO) donor and a nitrovasodilator drug used for patients with hypertensive crisis, has been shown to promote angiogenesis. However, direct evidence showing the involvement of NO in the SNP induced angiogenesis is not available. Accordingly, we assessed whether NO generated from SNP-stimulated ovine fetoplacental artery endothelial (OFPAE) cell proliferation via activation of mitogen-activated protein kinase 3/1 (MAPK3/1, also termed ERK1/2). We observed that SNP dose dependently stimulated (P < 0.05) cell proliferation with a maximal effect at 1 microM and that SNP rapidly ( 75% of PPHa. Only a small number of headaches (4%) were incapacitating. Postdural puncture headache accounted for 4.7% of all PPHa. Significant risk factors for the development of PPHa were: known inadvertent dural puncture [odds ratio (OR)adj = 6.36; 95% confidence interval (CI) 1.29, 31.24]; previous headache history (1-12/yr-OR(adj) = 1.57; 95% CI 1.01, 2.44; > 12/yr-OR(adj) = 2.25; 95% CI 1.63, 3.11); multiparity (OR(adj) = 1.37; 95% CI 1.03, 1.82) and increasing age (OR(adj) = 1.03/yr; 95% CI 1.00, 1.06). CONCLUSIONS: Postpartum headaches are common, often first noted after discharge from hospital. The majority are related to primary headache disorders. Increased awareness of this epidemiological relationship and improved diagnosis of primary headache conditions may lead to improved headache-specific therapy and avoidance of unnecessary investigations or read-mission to hospital. PMID- 16251566 TI - Esophageal Doppler and thermodilution are not interchangeable for determination of cardiac output. AB - PURPOSE: This study compares thermodilution cardiac output (TD-CO) and esophageal Doppler cardiac output (ED-CO) during periods of hemodynamic stability and after heart stabilization during off-pump coronary artery bypass (OPCAB) surgery. METHODS: After Institutional Review Board approval, 58 patients undergoing OPCAB had simultaneous comparison of TD-CO and ED-CO at three time periods. Measurements were recorded, in a blinded manner, after probe insertion (T0), immediately before and after (T1,T2) heart displacement and before starting any pharmacological treatment (if needed) to maintain systolic blood pressure to its value before heart mobilization. Measurements were also taken before sternal closure (Tfinal). RESULTS: Three hundred and two pairs of data were analyzed using the Bland and Altman method. Bias, standard deviation (SD) of the bias (precision), and degree of agreement (bias +/- 2 SD) were calculated. Based on published literature, we considered that the highest degree of agreement should be < 0.5 L.min(-1) to consider both methods as interchangeable. At T0, bias and SD of bias between TD-CO and ED-CO were -0.1 +/- 1.0 L.min(-1). Immediately before heart stabilization, bias +/- SD was 0.6 +/- 1.0 L.min(-1) and after heart displacement, 0.5 +/- 0.8 L.min(-1). At Tfinal, bias +/- SD was 0.7+/- 0.7 L.min( 1). CONCLUSION: Because the degree of agreement was > 0.5 L.min(-1) at all measurement periods except T0, we conclude that TD and ED are not interchangeable at any time during OPCAB surgery. PMID- 16251567 TI - Lung mechanics and gas exchange in one-lung ventilation following contralateral resection. AB - PURPOSE: To describe the anesthetic management of a patient with previous left lower lobe resection who was submitted to a right upper lobectomy and review the changes in gas exchange and respiratory mechanics which occurred intraoperatively. CLINICAL FEATURES: A 69-yr-old male with lung cancer, emphysema and obstructive sleep apnea, presented for a right upper lobectomy. His history was also positive for a left lower lobectomy six years previously. Intraoperative lung isolation was achieved using a 41 F left double-lumen tube (DLT). Monitoring the respiratory mechanics allowed for continuous adjustment of ventilator settings during the various phases of the surgery avoiding the risks of barotrauma and volutrauma. Problems with oxygenation occurred during one-lung ventilation. CONCLUSION: This case report shows that a severe level of hypoxemia and hypercarbia associated to lung mechanical property changes can be observed during the OLV phase. Application of continuous positive airway pressure on the non-dependent lung partially corrected blood oxygenation. Lobe isolation techniques should be considered as useful options for intraoperative airway management for these patients. PMID- 16251568 TI - Congenital tracheal stenosis in a boy with Rubinstein-Taybi syndrome. PMID- 16251569 TI - Success of the Cobra after failure of the Fastrach in the difficult airway. PMID- 16251570 TI - "The critical airway". PMID- 16251571 TI - Anesthetic management of bronchopleurocutaneous fistula--an alternate approach. PMID- 16251572 TI - "Keep out of trouble" airway algorithm. PMID- 16251573 TI - Bilateral foot drop: looking for the needle in the wrong haystack? PMID- 16251574 TI - Does bilateral thoracic sympathectomy predispose to reflex bronchospasm following tracheal intubation? PMID- 16251575 TI - Retained and cut stimulating infraclavicular catheter. PMID- 16251576 TI - Effectiveness of bolus landiolol on paroxysmal atrial tachycardia. PMID- 16251577 TI - Mean arterial blood pressure estimation and its limitation. PMID- 16251578 TI - Continuous renal replacement therapy. PMID- 16251579 TI - Sulfadiazine-induced methemoglobinemia in a boy with thalassemia. PMID- 16251580 TI - Marital quality, health, and aging: gender equity? AB - Recent research shows that poor marital quality adversely affects trajectories of physical health over time and that these adverse effects are similar for men and women. These studies test the possibility of gender differences in vulnerability to poor marital quality, but they fail to take into account possible gender differences in exposure to poor marital quality. We present longitudinal evidence to show that although the impact of marital quality on physical health trajectories may be similar for married men and women, generally lower levels of marital quality experienced by women may translate into a sustained disadvantage for the health of married women over the life course. These findings frame the call for renewed theoretical work on gender and marriage that takes into account both gender similarity in response to marital quality as well as gender differences in the experience of marriage over the life course. PMID- 16251581 TI - Family structure, gender, and health in the context of the life course. AB - We estimate the effects of single parenthood on parental health and determine whether such effects are similar for all single parents or whether there are variations by gender among young, middle-aged, and older adults. The results of our analyses of the Canadian National Population Health Survey (NPHS) reveal that single parenthood is associated with elevated psychological distress and alcohol consumption among women, especially among those who are in younger age groups. Although we find no such differences among men, there is some indication of elevated distress among younger fathers. We discuss the implications of these findings for thinking about single parenthood at various life stages. PMID- 16251582 TI - Processes of cumulative adversity: childhood disadvantage and increased risk of heart attack across the life course. AB - OBJECTIVES: This article examines how processes of cumulative adversity shape heart attack risk trajectories across the life course. METHODS: Our sample includes 9,760 Health and Retirement Study respondents born between 1931 and 1941. Using self-reported retrospective measures of respondents' early background, we first identify three latent classes with differential exposure to childhood disadvantage. Intervening covariates associated with educational attainment, employment status, income attainment, marital history, and health behaviors are added to capture sequential processes of adversity. Final latent class cluster models estimate the cumulative impact of these covariates on three different heart attack risk trajectories between 1992 and 2002: high, increasing, and low. RESULTS: Early disadvantage and childhood illness have severe enduring effects and increase the risk for heart attack. Adult pathways, however, differentially influence trajectories of heart attack risk and mediate the effects of early disadvantage. DISCUSSION: Findings suggest that future research should consider how processes of cumulative adversity initiated in childhood influence health outcomes in older ages. PMID- 16251583 TI - Early adversity and later health: the intergenerational transmission of adversity through mental disorder and physical illness. AB - OBJECTIVES: The authors' objective was to investigate processes that account for the transmission of socioeconomic adversity from one generation to the next through mental disorder and physical illness. METHODS: The present longitudinal study of 485 youth used structural equation models to test an intergenerational model proposing that: (a) stressful childhood experiences in the family of origin contribute to the development of mental disorder and physical illness during adolescence both directly and indirectly through disruption in an adolescent's transition to young adulthood; (b) during the transition to adulthood, mental disorders and physical illnesses increase in part through reciprocal influence; and (c) both the levels of and changes in mental disorder and physical illness are independently associated with adverse life circumstances during early adulthood. RESULTS: Findings generally supported the hypothesized model. Family of origin adversity contributed to the impaired mental and physical health of adolescents. This influence was largely mediated through adolescents' disrupted transition to young adulthood. Levels of both mental and physical illnesses independently contributed to young adult adversity. Levels of physical health problems influenced changes in mental disorders. Changes in both mental and physical illnesses are also associated with young adult adversity. DISCUSSION: The study demonstrates key mediating pathways in the intergenerational transmission of social adversity and also highlights the importance of improving both socioeconomic and health resources for adolescents. PMID- 16251584 TI - Conceptualizing and identifying cumulative adversity and protective resources: implications for understanding health inequalities. AB - This article focuses on cumulative adversity and protective resources, both social and biological, that interrupt or deflect individuals from optimal life course trajectories and contribute to widening gaps in health. Under the guiding framework of cumulative adversity and/or advantage, this narrative discusses the theoretical framework of cumulative adversity, presents identified sources of cumulative adversity and protective resources, and highlights the utilization of the life-course approach. Numerous social and biological adverse conditions are identified across multiple domains. Utilizing the life-course perspective in identifying early life determinants and the paucity of information regarding identified protective factors are discussed. Understanding health inequalities requires attention paid to heterogeneity in the impact of social statuses as well as sources of cumulative adversity and protective resources within diverging trajectories across the life course. Intervention implications are discussed, and suggestions for future research are made. PMID- 16251585 TI - Socioeconomic status and health across the life course: progress and prospects. PMID- 16251586 TI - Continuity and change in the social stratification of aging and health over the life course: evidence from a nationally representative longitudinal study from 1986 to 2001/2002 (Americans' Changing Lives Study). AB - OBJECTIVES: This article overviews previously published and ongoing research from the Americans' Changing Lives (ACL) Study, a longitudinal study of a nationally representative sample of 3,617 adults aged 25 years and older when first interviewed in 1986, focusing on socioeconomic disparities in the way health changes with age during middle and later life, especially in terms of compression of morbidity/functional limitations. METHODS: A variety of descriptive and multivariate regression and growth curve analyses are done on the ACL sample, now surveyed over four waves spanning 15.5 years between 1986 and 2001/2002 with continuing mortality ascertainment via the National Death Index, death certificate searches, and informant reports. RESULTS: Both cross-sectional and longitudinal analyses indicate that socioeconomic disparities in health are small in early adulthood, increase through middle and early old age, and then lessen again in later old age. In other terms, compression of morbidity/functional limitations into the later stages of the life course is realized to a much greater degree among the better educated compared with the less educated. Cross sectional evidence suggests that this reflects differential exposure to or experience of a wide range of psychosocial, environmental, and biomedical risk factors for health (and perhaps their differential impact at different ages and life stages), as well as variations in biological robustness and frailty and also perhaps in the strength of social welfare supports for health at different life stages. Longitudinal analyses reveal several new insights: (a) The flow of causality is much greater from socioeconomic position to health than vice versa; (b) education plays a greater role relative to income in the onset of functional limitations, whereas income has much stronger effects on their progression or course; and (c) educational disparities in the onset and hence of compression of functional limitations over the life course have increased strikingly in later middle and early old age (ages 55-84 years) since 1986. DISCUSSION: The results indicate that understanding and alleviating social disparities in health are both theoretically and methodologically quintessential problems of life course analysis and research. PMID- 16251587 TI - Controlling disease and creating disparities: a fundamental cause perspective. AB - The United States and other developed countries experienced enormous improvements in population health during the 20th century. In the context of this dramatic positive change, health disparities by race and socioeconomic status emerged for several potent killers. Any explanation for current health disparities must take these changing patterns into account. Any explanation that ignores large improvements in population health and fails to account for the emergence of disparities for specific diseases is an inadequate explanation of current disparities. We argue that genetic explanations and some prominent social causation explanations are incompatible with these facts. We propose that the theory of "fundamental causes" can account for both vast improvements in population health and the creation of large socioeconomic and racial disparities in mortality for specific causes of death over time. Specifically, we argue that it is our enormously expanded capacity to control disease and death in combination with existing social and economic inequalities that create health disparities by race and socioeconomic status: When we develop the ability to control disease and death, the benefits of this new-found ability are distributed according to resources of knowledge, money, power, prestige, and beneficial social connections. We present data on changing mortality patterns by race and socioeconomic status for two types of diseases: those for which our capacity to prevent death has increased significantly and those for which we remain largely unable to prevent death. Time trends in mortality patterns are consistent with the fundamental cause explanation. PMID- 16251588 TI - Do daily stress processes account for socioeconomic health disparities? AB - OBJECTIVES: The present study examined the extent to which daily stressor severity and appraisals of the stressors accounted for socioeconomic disparities in health. METHODS: Data from the National Study of Daily Experiences and the Midlife in the United States Survey were combined for the current analyses, resulting in 1,031 respondents who reported on 7,229 days. RESULTS: Respondents without a high school degree experienced more severe stressors and appraised stressors as posing greater risk to their financial situation and to their self concept than respondents with a high school or college degree. Differences in severity and stressor appraisal accounted for education differences in psychological distress and physical health symptoms. DISCUSSION: Findings suggest the importance of considering variation across stressors, particularly implications for self-concept, in understanding sources of differential stressor vulnerability. PMID- 16251589 TI - Rethinking gender differences in health: why we need to integrate social and biological perspectives. AB - The complexity of gender differences in health (i.e., men's lower life expectancy and women's greater morbidity) extends beyond notions of either social or biological disadvantage. Gaps remain in understanding the antecedents of such differences and the issues this paradox raises regarding the connections between social and biological processes. Our goals in this analytic essay are to make the case that gender differences in health matter and that understanding these differences requires an explanation of why rational people are not effective in making health a priority in their everyday lives. We describe some salient gender health differences in cardiovascular disease, immune function and disorders, and depression and indicate why neither social nor biological perspectives alone are sufficient to account for them. We consider the limitations of current models of socioeconomic and racial/ethnic health disparities to explain the puzzling gender differences in health. Finally, we discuss constrained choice, a key issue that is missing in the current understanding of these gender differences, and call on the social science community to work with biomedical researchers on the interdisciplinary work required to address the paradoxical differences in men's and women's health. PMID- 16251590 TI - Media portrayals and health inequalities: a case study of characterizations of Gene x Environment interactions. AB - OBJECTIVES: This article examines how genetic and environmental interactions associated with health inequalities are constructed and framed in the presentation of scientific research. METHODS: It uses the example of a major article about depression in a longitudinal study of young adults that appeared in Science in 2003. RESULTS: This portrayal of findings related to health inequalities uses a genetic lens that privileges genetic influences and diminishes environmental ones. DISCUSSION: The emphasis on the genetic side of Gene x Environment interactions can serve to deflect attention away from the important impact of social inequalities on health. PMID- 16251591 TI - The health of U.S. racial and ethnic populations. AB - This article provides an overview of racial and ethnic disparities in health in the United States. It describes limitations linked to the quality and method of presentation of the available data. It also considers the complex ways in which immigrant status, race, and SES combine to affect health and outlines important directions for research that would enhance our understanding of the ways in which social factors can lead to changes in health status. PMID- 16251592 TI - Proceedings of the conference: Health Inequalities Across the Life Course. June 2004, University Park, Pennsylvania, USA. PMID- 16251593 TI - Health inequalities among minority populations. AB - There are several challenges facing scholars studying health inequalities among minority populations. Primary among these challenges are developing adequate measures of social inequality and introducing appropriate strategies for eliminating health disparities. More research is sorely needed on both of these fronts as evidenced by the health paradox facing black, middle class men and women. This effort, however, can best move the study of health inequalities forward when juxtaposed against theoretical paradigms that embrace the complexity of the intersection of race, class, and gender. PMID- 16251594 TI - Aging, migration, and mortality: current status of research on the Hispanic paradox. AB - OBJECTIVE: We reviewed recent evidence on the apparent Hispanic mortality paradox. METHODS: Recent studies using vital statistics, national community surveys linked to the National Death Index, Medicare data linked to application records for social security cards maintained in the Social Security Administration NUDIMENT file, and mortality follow-up by regional studies are reviewed critically. RESULTS: Data based on vital statistics show the greatest mortality advantage compared with non-Hispanic Whites for all Hispanics combined. The advantage is greatest among older people. National Community Surveys linked to the National Death Index show a narrowing of the advantage, and one study suggests that the Mexican Origin mortality advantage can be attributed to selective return migration of less healthy immigrants to Mexico. The Medicare NUDIMENT data that avoid problems of other data sets also show an advantage in mortality among Hispanic elders, although the advantage is considerably lower than is found using the vital statistics method. DISCUSSION: Although some research has recently begun to question whether indeed all Hispanic groups enjoy a mortality advantage, the majority of the evidence continues to support a mortality advantage at a minimum among Mexican Americans and especially in old age, at least among men, which may provide partial, albeit indirect, support for a selective return migration or "salmon bias" effect. There is a need to further explore the existence of a selective return migration effect with expanded data bases that include more subjects from the various Hispanic origins. To date, the majority of the evidence continues to support the Hispanic paradox at least among people of Mexican origin and calls for additional attention to this interesting and highly important phenomenon. PMID- 16251595 TI - A life-span developmental perspective on social status and health. AB - This article presents a life-span developmental (LSD) perspective on the linkages between social status and health. The objective is to develop a conceptual framework that is useful in understanding why people are differentially exposed to risks of disease or protective factors and the social conditions that link the effects of risk and protective factors to the social environment over the life span. The discussion distinguishes between the complementary concepts of "life span," "life cycle," and "life course," critical theoretical distinctions that may help refine hypotheses about the relationship between health and social status. We argue that life-cycle and life-course concepts can be viewed as embedded in a more general LSD perspective. Using the theoretical principles derived from this perspective, the review examines (a) gender differences, (b) race-ethnic experiences, (c) childhood experiences, (d) educational levels, (e) socioeconomic differences, and (f) age differences. The emphasis in the review is to highlight the value of a broader LSD perspective in the study of health inequalities. The article ends with a brief summary of where future research is headed and novel developments in the study of social status and health. PMID- 16251596 TI - Economic status over the life course and racial disparities in health. AB - OBJECTIVES: Racial and socioeconomic disparities in health have become a prominent feature of American society, though our understanding of the processes leading to such persistent disparities is still relatively limited. In this study, we focus on the impact of social and economic advantages and disadvantages over the life course on health disparities at older ages. In particular, we look at the roles of both cumulative and current financial resources and financial strains as determinants of a range of subjective and objective health assessments of physical conditions, functional impairment, and mental health. METHODS: Our data come from the 2001 Aging, Stress, and Health Study, which interviewed over 1,100 White and African American adults aged 65 years and older living in the Washington, D.C., metropolitan area. RESULTS AND DISCUSSION: Our results show that although racial and socioeconomic disparities in health do not follow a simple explanation, we do provide strong support for the fundamental importance of social and economic resources. Unlike previous studies that emphasize the role of financial resources such as income and wealth, we show that the lack of these resources, as indicated by high levels of financial strain, provides an important clue to how economic resources influence health. PMID- 16251597 TI - Social status and risky health behaviors: results from the health and retirement study. AB - OBJECTIVES: We focus on a hypothesized mechanism that may underlie the well documented link between social status and health-behavioral health risks. METHODS: We use longitudinal data from representative samples of 6,106 middle aged and 3,636 older adults from the Health and Retirement Study to examine the relationships between social status-including early life social status (e.g., parental schooling), ascribed social status (e.g., sex, race-ethnicity), and achieved social status (e.g., schooling, economic resources)-and behavioral health risks (e.g., weight, smoking, drinking, physical activity) to (1) assess how early life and ascribed social statuses are linked to behavioral health risks, (2) investigate the role of achieved factors in behavioral health risks, (3) test whether achieved status explains the contributions of early life and ascribed status, and (4) examine whether the social status and health risk relationships differ at midlife and older age. RESULTS: We find that early life, achieved, and ascribed social statuses strongly predict behavioral health risks, although the effects are stronger in midlife than they are in older age. DISCUSSION: Ascribed social statuses (and interactions of sex and race ethnicity), which are important predictors of behavioral health risks even net of early life and achieved social status, should be explored in future research. PMID- 16251598 TI - Mental health disparities across education and sex: a prospective analysis examining how they persist over the life course. AB - OBJECTIVES: Higher levels of psychopathology among people with lower socioeconomic status and among women persist as cohorts age. In this analysis, we examine whether the persistence of these disparities as a cohort ages results from (a) a single set of people within a disadvantaged group who have chronic psychopathology or (b) continually changing sets of people within a disadvantaged group who have psychopathology of short duration. METHODS: Data for this analysis come from the Epidemiologic Catchment Area Study, which includes two psychological assessments (depressive syndrome and psychological distress) in a population sample of adults in Baltimore, Maryland, collected 13 years apart. RESULTS AND DISCUSSION: Results indicate that the persistence of disparities across education resulted primarily from one single set of respondents with chronic psychopathology over the 13 years of the survey, while the persistence of disparities across sex involved new sets of women as the cohort aged. We discuss implications of these results for theory and policy. PMID- 16251599 TI - Gender disparities in health: strategic selection, careers, and cycles of control. AB - This article proposes a dynamic model of the intersections between gender, health, and the life course incorporating processes of strategic selection--of roles, relationships, and behavior. Men and women make decisions within a tangled web of multilayered, often contradictory, and frequently outdated institutional contexts of opportunity and constraint. Both their decisions and the institutions shaping them reflect prior as well as ongoing socialization and allocation mechanisms. These institutionalized scripts and regimes tend to reproduce gendered biographical paths around two central life foci: paid work (or careers) and unpaid family work (or careers). The gendered nature of occupational and family-care paths, in turn, produces patterned disparities in a constellation of health-related resources, relationships, and risks, as well as feelings of mastery and control. We call for research charting alternative constellations of these gendered health careers, their antecedents, temporal patterning, and consequences. PMID- 16251600 TI - Comparative aspects of lipid metabolism: impact on contemporary research and use of animal models. AB - The emerging obesity crisis and consequent concerns for corrective measures and appropriate public policy have stimulated research into causes, prevention, remediation, and health consequences of obesity and associated maladies. Such research areas include eating behavior, appetite control, and food intake regulation as well as the regulation of lipid metabolism, cardiovascular function, endocrine function, and dyslipidemia states utilizing various animal models and cell culture systems. Although the liver has a central role in lipid/fatty acid synthesis and glucose is the precursor for de novo fatty acid synthesis in rodents and humans, in many other species, adipose tissues are the primary sites of lipogenesis. In addition, many species utilize acetic acid as a precursor for fatty acid synthesis. This fundamental difference in the site of fatty acid synthesis and the pattern of consequent lipid trafficking influences overall animal lipid metabolism and the role of regulatory hormones and transcription factors. Researchers utilizing various animal species in targeted biomedical research should be aware of these species differences when interpreting their data. In addition, many animal species are used for food production, recreational, and companion purposes. Understanding the lipid metabolism regulatory mechanisms of such species from a comparative perspective is important for the proper nutrition and health of these animals. PMID- 16251601 TI - Fatty acid regulation of hepatic gene transcription. AB - Dietary fat regulates gene expression by controlling the activity or abundance of key transcription factors. In vitro binding and cell culture studies have identified many transcription factors as prospective targets for fatty acid regulation, including peroxisome proliferator-activated receptors (PPARalpha, beta, gamma1, and gamma2), sterol regulatory element binding protein-1c (SREBP 1c), hepatic nuclear factors (HNF-4alpha and gamma), retinoid X receptor (RXRalpha), liver X receptor (LXRalpha), and others. In vivo studies established that PPARalpha- and SREBP-1c-regulated genes are key targets for PUFA control of hepatic gene expression. PUFA activate PPARalpha by direct binding, leading to the induction of hepatic fatty acid oxidation. PUFA inhibit hepatic fatty acid synthesis by suppressing SREBP-1c nuclear abundance through several mechanisms, including suppression of SREBP-1c gene transcription and enhancement of proteasomal degradation and mRNA(SREBP1c) decay. Changes in intracellular nonesterified fatty acids (NEFA) correlate well with changes in PPARalpha activity and mRNA(SREBP-1c) abundance. Several mechanisms regulate intracellular NEFA composition, including fatty acid transport, acyl CoA synthetases and thioesterases, fatty acid elongases and desaturases, neutral and polar lipid lipases, and fatty acid oxidation. Many of these mechanisms are regulated by PPARalpha or SREBP-1c. Together, these mechanisms control hepatic lipid composition and affect whole-body lipid composition. PMID- 16251602 TI - Astaxanthin diminishes gap junctional intercellular communication in primary human fibroblasts. AB - Astaxanthin is a carotenoid found in plants and algae; it provides the color of marine seafood such as salmon, lobster, or shrimp. Carotenoids are antioxidants and exhibit other biological functions, including effects on gap junctional communication important for homeostasis, growth control, and development of cells. Cancer cells have an impaired gap junctional intercellular communication. The objective of the present study was to determine the effects of astaxanthin and canthaxanthin on gap junctional intercellular communication in vitro. Primary human skin fibroblasts were exposed to carotenoids from 0.001 to 10 micromol/L, and gap junctional communication was measured with a dye transfer assay. After incubation with canthaxanthin for 24 and 72 h, intercellular communication increased, whereas it was strongly diminished by astaxanthin at levels > 0.1 micromol/L. Inhibition was reversed when astaxanthin was withdrawn. Western blot analysis showed that after exposure to canthaxanthin, the amount of the gap junction protein connexin43 was increased. Incubation with astaxanthin led to a change in the phosphorylation pattern of connexin43, shifting from higher to lower phosphorylation states. We suggest that astaxanthin affects channel function by changing the phosphorylation pattern of connexin43. PMID- 16251603 TI - Megalin-mediated reuptake of retinol in the kidneys of mice is essential for vitamin A homeostasis. AB - The reuptake of retinol (ROH) and retinol-binding protein (RBP) in the kidneys is mediated by the endocytic receptor megalin, suggesting an important role for this receptor in vitamin A (VA) metabolism. We examined the extent to which megalin deficiency may affect urinary ROH excretion, levels of ROH and RBP in plasma, as well as storage of VA in liver and kidney. For this purpose, mice with a kidney specific megalin gene defect (megalin(lox/lox); apoE(Cre)) and control mice (megalin(lox/lox)) were fed either a basal diet containing 4500 retinol equivalents (RE)/kg diet or a diet without VA during experimental periods of 42 and 84 d. Urinary ROH excretion was observed only in megalin(lox/lox); apoE(Cre) mice (P < 0.0001, 2-way ANOVA) and not in the controls. Plasma ROH and RBP differed only by diet (P < 0.05), but not genotype (P = 0.615). A major effect of megalin deficiency, however, was evident in retinyl ester levels in the liver (P < 0.05), which were approximately 37% lower than those in megalin(lox/lox) controls (P < 0.05, Student's t test) during the 84-d period of dietary VA deprivation. Kidney levels of VA were not affected by the receptor gene defect. The findings demonstrate that urinary ROH excretion caused by megalin deficiency requires accelerated mobilization of hepatic VA stores to maintain normal plasma ROH levels, which suggests that megalin plays an essential role in systemic VA homeostasis. PMID- 16251604 TI - Reduced adiposity in bitter melon (Momordica charantia)-fed rats is associated with increased lipid oxidative enzyme activities and uncoupling protein expression. AB - To further explore the antiobesity effect of freeze-dried bitter melon (BM) juice, activities of mitochondrial lipid oxidative enzymes as well as the expression of uncoupling proteins and their transcription coactivator peroxisome proliferator-activated receptor-gamma coactivator-1 alpha (PGC-1alpha) were determined in diet-induced obese (DIO) rats. Rats were fed high-fat (HF) diets to induce obesity, and the effect of BM was assessed at doses of 0.75, 1.0, or 1.25% (wt:wt). In a dose-response experiment, BM-supplemented rats had lower energy efficiency (g weight gained/kJ consumed), visceral fat mass, serum glucose, and insulin resistance index, but higher plasma norepinephrine than unsupplemented rats (P < 0.05). Hepatic and skeletal muscle triglyceride concentrations were lower in supplemented HF diet-fed rats than in unsupplemented HF diet-fed rats (P < 0.05). An HF diet supplemented with BM elevated activities of hepatic and muscle mitochondrial carnitine palmitoyl transferase-I (CPT-I) and acyl-CoA dehydrogenase (AD) (P < 0.05). In another experiment, BM (1.0 g/100 g) lowered visceral fat mass but increased serum adiponectin concentration in HF diet-fed rats (P < 0.05). In the final study, rats were fed the HF diet with 0, 1.0 or 1.25% BM. Both groups of BM-supplemented rats had higher uncoupling protein 1 in brown adipose tissue (P < 0.05) and uncoupling protein 3 in red gastrocnemius muscle (P < 0.05), measured by Western blotting and RT-PCR, than the controls. The expression of the transcription coactivator PGC-1alpha in both tissues was also significantly elevated in the BM-supplemented rats (P < 0.05). The present results suggest that decreased adiposity in BM-supplemented rats may result from lower metabolic efficiency, a consequence of increased lipid oxidation and mitochondrial uncoupling. PMID- 16251605 TI - Proteomic analysis reveals changes in the liver protein pattern of rats exposed to dietary folate deficiency. AB - Epidemiologic and experimental studies showed that folate deficiency is associated with increased risk of degenerative diseases by enhancing abnormal one carbon metabolism. We studied the changes in the proteome of liver, the main tissue of folate storage and metabolism, in a rat model of dietary folate depletion. Four-month-old rats were fed for 4 wk an amino acid-defined diet without folate and compared with pair-fed rats given the same diet adequately supplemented with folic acid. Folate deprivation decreased plasma and hepatic folate concentrations dramatically, while increasing homocysteinemia significantly. Using 2-dimensional electrophoresis and matrix-assisted laser desorption/ionization time-of-flight MS, we identified 9 spots corresponding to differentially expressed proteins in the liver of folate-deficient rats compared with controls. Among those spots, 4 had a significantly increased volume, whereas the volume of the 5 other spots was decreased. Upregulated proteins included glutathione peroxidase (GPx) 1 and peroxiredoxin 6, 2 enzymes involved in the response to oxidative stress, and MAWD binding protein (MAWDBP), which has been associated with cancer. MAWDBP was simultaneously identified as a second spot with a lower isoelectric point (pI) that vanished almost completely after folate deficiency. Decreased abundance was also observed for cofilin 1, a protein linked to tumorigenesis, and for the GRP 75 precursor and preproalbumin, both of which are responsive to oxidative stress and/or inflammation. Moreover, an enzyme activity assay and/or Western blot analysis of GPx-1 and MAWDBP confirmed the proteomic findings. Our results show that folate deficiency modifies the abundance of several liver proteins consistently with adaptive tissue responses to oxidative and degenerative processes. PMID- 16251606 TI - Scurvy leads to endoplasmic reticulum stress and apoptosis in the liver of Guinea pigs. AB - Insufficient ascorbate intake causes scurvy in certain species. Beyond its known functions, it has been suggested that ascorbate participates in oxidative protein folding in the endoplasmic reticulum (ER). Because redox imbalance in this organelle might cause ER stress and apoptosis, we hypothesized that this might contribute to the pathology of scurvy. Guinea pigs were divided into 7 groups: the control group was fed a commercial guinea pig food containing 0.1 g/100 g ascorbate for 4 wk, 5 groups consumed an ascorbate-free food for 0, 1, 2, 3, or 4 wk and 1 group was fed this scorbutic diet for 2 wk and then the commercial food plus 1 g/L ascorbate in drinking water for 2 wk. TBARS generation and the expression of some ER chaperones and foldases were determined in hepatic microsomes. The apoptotic index was assessed in histological sections. Although ascorbate, measured by HPLC, was undetectable in the livers of the guinea pigs after they had consumed the scorbutic diet for 2 wk, the microsomal TBARS level was elevated relative to the initial value only at wk 4. Western blot revealed the induction of GRP78, GRP94, and protein disulfide isomerase at wk 3 and 4. Apoptosis was greater than in the control, beginning at wk 3. None of the alterations occurred in the groups fed the commercial guinea pig food or ascorbate-free food followed by ascorbate supplementation. Therefore, persistent ascorbate deficiency leads to ER stress, unfolded protein response, and apoptosis in the liver, suggesting that insufficient protein processing participates in the pathology of scurvy. PMID- 16251607 TI - The apolipoprotein E gene promoter (-219G/T) polymorphism determines insulin sensitivity in response to dietary fat in healthy young adults. AB - Insulin sensitivity (IS) is determined by genetic and environmental factors, including diet. The apoE gene promoter -219G/T polymorphism is associated with coronary heart disease and increased postprandial triacylglycerol-rich lipoprotein concentration, circumstances related to insulin resistance. Thus, our aim was to determine whether this polymorphism modified the IS response to dietary fat in healthy young adults. Volunteers (n = 43) with the apoE3/E3 genotype (8 GG, 25 GT and 10 TT) completed 3 dietary periods, each lasting 4 wk. They first consumed a SFA-rich diet [38% fat (% of energy in the total diet), 20% SFA (% of energy in the total diet)], and then, in a randomized, crossover design, a carbohydrate (CHO)-rich diet (30% fat, 55% CHO) or a monounsaturated fatty acid (MUFA)-rich diet (38% fat, 22% MUFA). After each diet period, we investigated peripheral IS using the insulin suppression test. The steady-state plasma glucose (SSPG) concentration was lower (P < 0.05) in GG subjects than in GT and TT individuals, regardless of the diet consumed. Significant diet x genotype interactions were found for SSPG and plasma nonesterified FFA (NEFA) concentrations. Thus, the shift from the SFA-rich diet to the MUFA- or CHO-rich diets decreased (P < 0.05) the SSPG and NEFA concentrations in GG and GT, but not in TT subjects. In conclusion, carriers of the -219T allele are less insulin sensitive than GG individuals. Furthermore, only carriers of the -219G allele have improved IS when MUFA- or CHO-rich diets are consumed instead of a SFA-rich diet. PMID- 16251608 TI - Long-chain polyunsaturated fatty acids upregulate LDL receptor protein expression in fibroblasts and HepG2 cells. AB - The objective of this study was to investigate the effect of individual PUFAs on LDL receptor (LDLr) expression in human fibroblasts and HepG2 cells, and to evaluate whether acyl CoA:cholesterol acyltransferase (ACAT) and sterol regulatory element-binding protein 1 (SREBP-1) were involved in the regulation of LDLr expression by fatty acids. When fibroblasts and HepG2 cells were cultured with serum-free defined medium for 48 h, there was a 3- to 5-fold (P < 0.05) increase in LDLr protein and mRNA levels. Incubation of fibroblasts and HepG2 cells in serum-free medium supplemented with 25-hydroxycholesterol (25OH cholesterol, 5 mg/L) for 24 h decreased LDLr protein and mRNA levels by 50-90% (P < 0.05). Arachidonic acid [AA, 20:4(n-6)], EPA [20:5(n-3)], and DHA [22:6(n-3)] antagonized the depression of LDLr gene expression by 25OH-cholesterol and increased LDLr protein abundance 1- to 3-fold (P < 0.05), but had no significant effects on LDLr mRNA levels. Oleic (18:1), linoleic (18:2), and alpha-linolenic acids [18:3(n-3)] did not significantly affect LDLr expression. ACAT inhibitor (58-035, 1 mg/L) attenuated the regulatory effect of AA on LDLr protein abundance by approximately 40% (P < 0.05), but did not modify the regulatory effects of other unsaturated fatty acids in HepG2 cells. The present results suggest that AA, EPA, and DHA increase LDLr protein levels, and that ACAT plays a role in modulating the effects of AA on LDLr protein levels. Furthermore, the effects of the fatty acids appeared to be independent of any change in SREBP-1 protein. PMID- 16251609 TI - Vegetables affect the expression of genes involved in carcinogenic and anticarcinogenic processes in the lungs of female C57BL/6 mice. AB - Worldwide, lung cancer is the most prevalent and lethal malignant disease. In addition to avoidance of the most predominant risk factor, i.e., tobacco use, consumption of high amounts of vegetables and fruits could be an effective means of preventing lung cancer. However, the molecular mechanisms underlying lung cancer risk reduction by vegetables are not clear. In the present study, the effect of vegetables on gene expression changes in the lungs of female C57Bl/6 mice was investigated using cDNA microarray technology. The mice were fed 1 of 8 diets for 2 wk: a control diet containing no vegetables (diet 1); a diet containing a vegetable mixture at 100 (diet 2, 10% dose), 200 (diet 3, 20% dose), or 400 (diet 4, 40% dose) g/kg; or a diet containing cauliflower at 70 (diet 5, 7% dose); carrots at 73 (diet 6, 7.3% dose); peas at 226 (diet 7, 22.6% dose); or onions at 31 (diet 8, 3.1% dose) g/kg. The vegetable mixture consisted of these 4 individual vegetables. After the mice were killed, the lungs were removed and total RNA was isolated from the lungs for expression analysis of 602 genes involved in pathways of (anti)-carcinogenesis. The results of this study suggest that individual vegetables have a higher potential of modulating genes (5 from the 8 modulated genes) in favor of lung cancer risk prevention, in comparison with the vegetable mixture (2 from the 7 modulated genes); the other gene modulations are expected to enhance lung cancer risk. The pathways involved were miscellaneous and included cell growth, apoptosis, biotransformation, and immune response. Furthermore, carrots were able to modulate most gene expressions, and most of these effects occurred in processes that favored lung cancer risk prevention. The current study provides more insight into the genetic mechanisms by which vegetables, in particular carrots, can prevent lung cancer risk. PMID- 16251610 TI - Insulin-like growth factor-I (IGF-I) attenuates jejunal atrophy in association with increased expression of IGF-I binding protein-5 in parenterally fed mice. AB - Total parenteral nutrition (TPN) induces dramatic mucosal hypoplasia in rat small intestine that is attenuated by insulin-like growth factor-I (IGF-I). Our aim was to determine the extent of TPN-induced intestinal atrophy and its response to infusion of IGF-I in mice. Male C57BL/6 mice (18-22 g) were maintained with TPN, TPN plus co-infusion of recombinant human IGF-I [2.5 mg/(kg . d)] or oral feeding for 5 d. Body weights did not differ among the groups although serum IGF-I was increased by 78% with IGF-I infusion. IGF-I prevented the significant 25% reduction in mass of the intact small intestine due to TPN compared with oral feeding. Greater TPN-induced atrophy was noted in duodenum and jejunum than ileum. Jejunal atrophy induced by TPN reflected significant decreases in muscularis mass and concentrations of protein and DNA; mucosal cellularity was not altered by TPN. TPN induced a significant decrease in jejunal muscularis width that was reversed by IGF-I with no differences in mucosal villus height and crypt depth. Local expression of IGF-I binding protein (IGFBP)-5 positively modulates the intestinotrophic effects of IGF-I. Jejunal atrophy due to TPN and growth due to IGF-I were directly associated with expression of IGFBP-5 mRNA. TPN decreased IGFBP-5 mRNA by 60% and IGF-I increased IGFBP-5 mRNA by 200% with no change in IGF-I mRNA compared with oral feeding. In summary, TPN induces significant 25% atrophy of the mouse small intestine that is attenuated by IGF-I in association with increased expression of IGFBP-5. Compared with rats, TPN induced atrophy is less severe and occurs primarily in the jejunal muscularis layer in mice. PMID- 16251611 TI - Garlic organosulfur compounds upregulate the expression of the pi class of glutathione S-transferase in rat primary hepatocytes. AB - The chemopreventive property of garlic is related in part to its induction of phase II detoxification enzymes. In the present study, we investigated the modulatory effect of 3 garlic organosulfur compounds, i.e., diallyl sulfide (DAS), diallyl disulfide (DADS), and diallyl trisulfide (DATS), which differ in their number of sulfur atoms, on the gene expression of the pi class of glutathione S-transferase (GSTP). Hepatocytes isolated from male Sprague-Dawley rats were cultured with 50-200 micromol/L of DAS, DADS, or DATS for 24 h. DADS and DATS increased GST activity toward ethacrynic acid by 40 and 66%, respectively (P < 0.05). Moreover, both garlic allyl sulfides dose dependently induced GSTP mRNA and protein expression. DATS increased the protein level more than DADS (P < 0.05). In contrast, DAS did not affect the activity or the protein or mRNA levels of this phase II drug-metabolizing enzyme. In Clone 9 liver cells, the pTA-luciferase reporter assay showed that luciferase activity in DADS- and DATS-treated cells was 2.8- and 3.9-fold higher than that in control cells, respectively (P < 0.05). Again, luciferase activity was not affected by treatment with DAS. Deletion of -2.7 to -2.6 kb in the GSTP promoter region, which contains the GSTP enhancer (GPE) I element, abolished the upregulation of GSTP transcription by DADS and DATS. Deletion of GPE II, however, did not affect the induction of reporter activity. In conclusion, the effectiveness of 3 garlic allyl sulfides on GSTP expression was related to the number of sulfur atoms in the molecules, and GPE I was responsible for this upregulation. PMID- 16251612 TI - Arachidonic acid predominates in the membrane phosphoglycerides of the early and term human placenta. AB - The aim of this study was to determine whether the high concentration of arachidonic acid (AA) in term placentae accumulates during pregnancy or is an inherent characteristic of placental lipids. We investigated the lipid content and fatty acid composition of the human placental phospholipids at 2 gestational periods, early in pregnancy (8-14 wk, n = 48) and at term (38-41 wk of gestation, n = 19). The subjects were healthy, normotensive, and free of medical and obstetric complications. The lipid concentration of placentae increased from 0.8% in early gestation to 1.4% at term (P < 0.0001). The mean proportions of AA were lower in the choline (P < 0.05), inositol (P < 0.0001), and ethanolamine (P < 0.0001) phosphoglycerides of the term compared with the early placenta. In contrast, the proportions of the immediate precursor of AA, dihomo-gamma linolenic acid (DGLA), were higher in the term placenta, particularly in the inositol and serine phosphoglycerides (P < 0.0001). In sphingomyelin, the percentage of lignoceric acid was increased and that of nervonic acid was reduced at term (P < 0.01). The dominance of AA, particularly in the early placenta, suggests that it has an important role for placental development, i.e., organogenesis and vascularization. There was no evidence of an accumulation of AA in the placenta toward term, which might be a trigger for parturition. In contrast, the increased proportion of DGLA (precursor of the vasorelaxant and anticoagulant prostaglandin E(1)) at term is more consistent with a profile favoring optimal blood flow to nourish the fetal growth spurt. PMID- 16251613 TI - Iron deficiency anemia and depleted body iron reserves are prevalent among pregnant African-American adolescents. AB - Anemia is prevalent among pregnant adolescents, but few data exist on biochemical indicators of iron status in this group. We hypothesized that among an at-risk population of African-American, pregnant adolescents, the degree of iron depletion and deficiency would be marked, and that iron deficiency anemia would comprise the majority of the observed anemia. To examine this, blood samples were collected from 80 girls (< or =18 y old) attending an inner city maternity clinic, 23 of whom were studied longitudinally in the 2nd and 3rd trimesters depending on contact at the clinic. Sample sizes for the biomarkers varied according to the blood volume available at the time the assays were completed. Descriptive statistics were applied to characterize iron status, and multivariate regression and logistic analyses were used to identify significant determinants of iron status. Depleted iron stores (ferritin < or = 15 microg/L) were indicated for 25% (n = 44) and 61% (n = 59) of adolescents during the 2nd and 3rd trimesters, respectively. Serum folate (39.3 +/- 15.4 nmol/L, n = 60), RBC folate (2378 +/- 971 nmol/L, n = 60), and serum vitamin B-12 concentrations (313 +/- 163 pmol/L, n = 60) were within normal ranges. Adolescents with serum transferrin receptor:serum ferritin ratios (R:F ratio) > 300 during the 2nd trimester were 12.5 times (95% CI 2.83, 55.25) more likely to be classified with iron deficiency anemia during the 3rd trimester (P = 0.0002) than those with lower ratios. Estimates of body iron were lower in those tested after wk 26 of gestation (P < 0.0001), and reserves were depleted in 5.0% vs. 31.3% of the 2nd (n = 40) and 3rd (n = 48) trimester cohorts, respectively. In conclusion, iron-deficiency anemia was prevalent among these pregnant minority adolescents. Targeted screening and interventions to improve diet and compliance with prenatal iron supplementation are warranted for this at-risk group. PMID- 16251614 TI - Calcium absorption from corn tortilla is relatively high and is dependent upon calcium content and liming in Mexican women. AB - Corn tortillas are the staple food of Mexico. During their preparation, calcium is added to the tortillas; therefore, tortillas are the main source of calcium for a large proportion of the population. The bioavailability of calcium from lime-treated tortillas in humans is not known. The objectives of the present study were to determine calcium absorption from corn tortilla, to determine the effect of lime treatment on calcium absorption from corn tortilla, and to compare calcium absorption from tortilla prepared with a commercial corn flour and tortillas prepared with the traditional lime treatment at home. Nonpregnant, nonlactating women (n = 9) were administered 3 different treatments: 1) 180 g of corn tortilla prepared from corn flour with no lime treatment (CF), 2) 180 g of corn tortilla prepared from lime-treated commercial corn flour (LTCCF), or 3) 180 g of corn tortillas prepared from lime-treated home-prepared corn flour (LTHCF). Calcium absorption was measured using an established dual-tracer stable isotope technique. Calcium absorption of CF, LTCCF, and LTHCF was (mean +/- SD): 44 +/- 3.2, 32 +/- 4.4, and 30 +/- 2.4%, respectively; the fractional calcium absorption from CF differed from that of either LTCCF or LTHCF (P < 0.01). The total amount of calcium absorbed per treatment was higher in LTHCF (98.10 mg +/- 21.7) than in LTCCF (59.9 mg +/- 23.7, P < 0.001) and CF (3.78 mg +/- 0.9, P < 0.0.001). In conclusion, calcium absorption from corn tortillas is high and dependent on calcium concentration. The addition of calcium during lime treatment increases calcium concentration and total calcium absorption. PMID- 16251615 TI - Anthocyanins exist in the circulation primarily as metabolites in adult men. AB - Anthocyanins are reported to have many "health promoting" properties; however, despite numerous reports of their bioactivities, their absorption and metabolism in humans are poorly understood. The objective of this research was to detail the pharmacokinetic parameters of anthocyanins after the administration of a 721-mg oral dose of cyanidin 3-glycosides from chokeberry extract to human subjects. Solid-phase extraction, preparative-HPLC, preparative-TLC, HPLC-diode array detection, HPLC-MS, and NMR were utilized to isolate, identity, and quantify anthocyanins in 0- to 7-h (0, 1, 2, 3, 4, 5, 6, 7 h) serum and 0- to 24-h urine samples (total individual urine voids over 24 h). The cumulative concentration of total anthocyanins (parent and metabolites) detected in the serum (0-7 h) was 376.65 +/- 16.20 (nmol x h)/L (area under the concentration time curve), reaching a maximum concentration (C(max) = 96.08 +/- 6.04 nmol/L) within 2.8 h. The parent anthocyanins represented only 32.0% [120.63 +/- 2.85 (nmol x h)/L] of the total anthocyanins detected with 68.0% [256.02 +/- 5.23 (nmol x h) identified as conjugated metabolites. Additionally, the total urinary excretion of anthocyanins over 24 h was 1071.54 +/- 375.46 microg, reaching a maximal rate of excretion (R(max) = 202.74 +/- 85.06 microg/h) at 3.72 +/- 0.83 h. Parallel to the serum data, only 32.5% (347.85 +/- 60.61 microg) of the anthocyanins excreted in the urine (total 24 h) were the parent compounds with 67.5% (723.69 +/- 92.59 microg) occurring as conjugated metabolites. The metabolites were identified as glucuronidated and methylated derivatives of the parent cyanidin 3-glycosides. The above results indicate that cyanidin 3-glycosides are rapidly absorbed and metabolized extensively following a moderate-to-high oral dose in humans. PMID- 16251616 TI - Breast-feeding influences cognitive development in Filipino children. AB - The importance of breast-feeding (BF) for cognitive development has been researched widely over the past several decades. Although scholars agree that children who breast-feed are generally more intelligent, it is uncertain whether this advantage is due to BF effects or to other accompanying healthy characteristics of women who breast-feed. This is a problem in nearly every study, and even in studies controlling for known confounding variables, residual confounding remains a concern. This study tried a new approach, evaluating the relation between BF and cognitive development or ability in a population in which BF was inversely correlated with socioeconomic advantages and other healthy maternal behaviors. Normal birthweight (NBW, n = 1790) and low birthweight (LBW, n = 189) (<2500 g) infants born in 1983-84 in Metropolitan Cebu, Philippines were followed from birth through middle childhood. Cognitive ability was assessed at ages 8.5 and 11.5 y with the Philippines Nonverbal Intelligence Test. Multivariable linear regressions were created to estimate crude and adjusted relations of various BF measures and later cognitive ability. After controlling for confounding variables, scores at 8.5 y were higher for infants breast-fed longer (1.6 points and 9.8 points higher among NBW and LBW infants, respectively, breast-fed for 12 to <18 mo vs. <6 mo). BF coefficients in both NBW and LBW 11.5 y models were attenuated but remained positive. This analysis highlights the importance of long-term BF after initial introduction of complementary foods, particularly in LBW infants born close to term. PMID- 16251617 TI - The use of NaFeEDTA-fortified fish sauce is an effective tool for controlling iron deficiency in women of childbearing age in rural Vietnam. AB - One cost-effective strategy for controlling iron deficiency is the fortification of staple foods or condiments with iron. We evaluated the effectiveness of fortifying fish sauce with NaFeEDTA for improving iron status in women of childbearing age in Vietnam in a double-blind intervention with randomization by village. All families in the selected villages were supplied with fish sauce that was either unfortified (Group C, 10 villages) or fortified with NaFeEDTA [9 mmol (500 mg) Fe/L, Group F, 11 villages] for 18 mo. The effect of fortification was assessed in the 576 women (n = 288/group) by measuring hemoglobin and serum ferritin (SF) at 6, 12, and 18 mo. Analysis of the group x time interaction using a repeated-measures test for each response demonstrated a significant effect of fortification on hemoglobin (P = 0.039) and log SF (P < 0.0001) in Group F with no significant changes in Group C. The prevalence of iron deficiency (SF < 12 microg/L) decreased from 22.3 to 4.0% and the prevalence of anemia (hemoglobin < 120 g/L) from 24.7 to 8.5% in Group F with no significant changes in Group C. NaFeEDTA fortification of fish sauce is an effective method for reducing the prevalence of iron deficiency in women in Vietnam. PMID- 16251618 TI - Season and ethnicity are determinants of serum 25-hydroxyvitamin D concentrations in New Zealand children aged 5-14 y. AB - New Zealand children, particularly those of Maori and Pacific ethnicity, may be at risk for low vitamin D status because of low vitamin D intakes, the country's latitude (35-46 degrees S), and skin color. The aim of this study was to determine 25-hydroxyvitamin D concentrations and their determinants in a national sample of New Zealand children aged 5-14 y. The 2002 National Children's Nutrition Survey was designed to survey New Zealand children, including oversampling of Maori and Pacific children to allow ethnic-specific analyses. A 2 stage recruitment process occurred using a random selection of schools, and children within each school. Serum 25-hydroxyvitamin D concentration [mean (99% CI) nmol/L] in Maori children (n = 456) was 43 (38,49), in Pacific (n = 646) 36 (31,42), and in New Zealand European and Others (NZEO) (n = 483) 53 (47,59). Among Maori, Pacific, and NZEO, the prevalence (%, 99% CI) of serum 25 hydroxyvitamin D deficiency (<17.5 nmol/L) was 5 (2,12), 8 (5,14), and 3 (1,7), respectively. The prevalence of insufficiency (<37.5 nmol/L) was 41 (29,53), 59 (42,75), and 25 (15,35), respectively. Multiple regression analysis found that 25 hydroxyvitamin D concentrations were lower in winter than summer [adjusted mean difference (99% CI) nmol/L; 15 (8,22)], lower in girls than boys [5 (1,10)], and lower in obese children than in those of "normal" weight [6 (1,11)]. Relative to NZEO, 25-hydroxyvitamin D concentrations were lower in Maori [9 (3,15)] and Pacific children [16 (10,22)]. Ethnicity and season are major determinants of serum 25-hydroxyvitamin D. There is a high prevalence of vitamin D insufficiency in New Zealand children, which may or may not contribute to increased risk of osteoporosis and other chronic disease. There is a pressing need for more convincing evidence concerning the health risks associated with the low vitamin D status in New Zealand children. PMID- 16251619 TI - The contribution of body protein to the supply of energy in starved newborn piglets is not preferentially suppressed by intravenous provision of glucose and fat. AB - Newborn piglets were used to study body protein preservation because it is critical to the survival of premature infants. Quantitative estimates of endogenous fuel use were obtained from 12 to 72 h of age in male piglets. Of the 40 piglets used (1300 +/- 205 g, mean +/- SD), 16 served as a 12-h-old body composition reference (R), 16 were starved (S) and received water only, and 8 received supplemental energy (E), obtaining 70% [210 kJ/(kg x d)] of their resting energy requirement as an i.v. mixture of glucose and Intralipid (65:35 energy ratio). Urine was collected continuously from the bladder via an umbilical urachal catheter. Total body water, glycogen, lipid, ash, and Kjeldahl-N were determined on whole-pig homogenates. Comparative slaughter was used to estimate the disappearance of body constituents of S and E pigs from 12 to 72 h of age. Midpoint body weight was used in these calculations. Supplemental energy decreased use of all body energy sources as indicated by the decrease in body dry matter disappearance, 41.6 +/- 8.8 vs. 25.5 +/- 5.9 g/kg (P = 0.0021) and protein (urinary N excretion), 995 +/- 508 vs. 329 +/- 135 mg/kg (P = 0.0119) over 60 h. Supplemental energy did not preferentially spare the percentage of the resting energy expenditure supplied by endogenous body protein (protein 37.6% +/- 9.6 vs. 41.7% +/- 10.4; lipid 25.7% +/- 5.2 vs. 20% +/- 4.1; glycogen 36.8% +/- 7.5 vs. 38.3% +/- 9.9; S vs. E) because it made up approximately 40% of the total in food deprived and supplemented piglets. PMID- 16251620 TI - Novel lycopene metabolites are detectable in plasma of preruminant calves after lycopene supplementation. AB - Appropriate animal models such as preruminant calves are necessary to study the complex physiological functions of carotenoids and to relate them to possible health effects in humans. In this study, the bioavailability and metabolism of lycopene from 2 dietary supplements were compared. LycoVit containing synthetic lycopene and Lyc-O-Mato containing natural tomato oleoresin were administered to 2 groups of preruminant calves (each n = 8) for 14 d in daily doses of 15 mg of lycopene. Plasma was analyzed for carotenoids before the intervention period, directly after, and each day for 5 d after the end of the intervention. All-trans and 5-cis lycopene, and 3 lycopene metabolites not previously found in calf plasma were detected. These metabolites contributed 52% of the total lycopene content measured at the end of the intervention period. Based on spectroscopic data, they might be hydrogenation products, which are formed from all-trans and/or 5-cis lycopene. In the LycoVit group, total lycopene concentrations were approximately 300% higher (286 +/- 89 nmol/L) than in the Lyc-O-Mato group (72 +/ 33 nmol/L) (P < 0.001). This indicates that, unlike in humans, lycopene from LycoVit and Lyc-O-Mato does not have equal bioavailabilities in preruminant calves. Therefore, the preruminant calf may not be a suitable animal model with which to study the biological and physiological effects of lycopene. PMID- 16251621 TI - Twice the amount of alpha-carotene isolated from carrots is as effective as beta carotene in maintaining the vitamin A status of Mongolian gerbils. AB - The vitamin A (VA) value of carotenoids from fruits and vegetables is affected by many factors. This study determined the VA value of alpha-carotene isolated from carrots compared with beta-carotene and retinyl acetate supplements fed to Mongolian gerbils (Meriones unguiculatus). Gerbils (n = 38) were fed a VA-free diet for 4 wk. At baseline, 6 gerbils were killed to determine liver VA. Gerbils were divided into 3 treatment groups (n = 9/group) and given 35, 35, or 17.5 nmol retinyl acetate, alpha-carotene or beta-carotene, respectively, in 2 divided doses 5 h apart each day. The remaining 5 gerbils received oil vehicle. Gerbils were killed after 3 wk of supplementation. Serum samples and livers were collected and analyzed for VA. Liver extracts were subsequently saponified to quantify alpha-retinol. Serum retinol concentrations did not differ among the groups. Liver retinyl palmitate concentrations were significantly higher in the retinyl acetate treatment group (0.198 +/- 0.051 micromol/g; P < 0.05) than in all other groups. The alpha- and beta-carotene treatments resulted in similar retinyl palmitate concentrations, i.e., 0.110 +/- 0.026 and 0.109 +/- 0.051 micromol/g, respectively, which did not differ from the concentrations in gerbils killed at baseline (0.123 +/- 0.024 micromol/g). The oil group had significantly less retinyl palmitate (0.061 +/- 0.029 micromol/g; P < 0.05) than all other groups. alpha-Retinol was detected in livers of the alpha-carotene group (0.062 +/- 0.013 micromol/g). Thus, twice the amount of purified alpha-carotene maintained VA status as well as beta-carotene in VA-depleted gerbils. Conversion factors were approximately 5.5 microg alpha-carotene or approximately 2.8 mug beta-carotene to 1 microg retinol. PMID- 16251622 TI - Selenium bioavailability from buckwheat bran in rats fed a modified AIN-93G torula yeast-based diet. AB - Selenium (Se) is an essential nutrient for humans and animals. The Se RDA for adult humans is 55 mug/d; however, dietary amounts as high as 200 mug/d in the highly available form of selenomethionine in yeast were shown to reduce the incidence of certain cancers. A number of natural foods contain relatively high amounts of Se; for the most part, however, the availability of food Se for absorption and utilization is unknown. This experiment was conducted to determine the bioavailability of Se from a high-protein, high-fiber bran-isolate of buckwheat groats that contains Se. The method used was based on the ability of Se from buckwheat bran to restore Se-dependent enzyme activities and tissue Se concentrations in Se-deficient rats. The responses produced from buckwheat bran Se were compared with a standard response curve generated by feeding graded amounts of Se as sodium selenite (Na(2)SeO(3); Na selenite) or selenomethionine (SeMet) in a newly reformulated AIN-93G-Torula yeast diet with a more balanced nutrient composition than older diets of this nature. Relative bioavailability was determined by using the slope-ratio assay method for enzyme data, or the parallel lines assay method for tissue Se concentration data. Results showed that Se availability from buckwheat bran based on the restoration of plasma Se was 70 80% as high as Na selenite or SeMet. However, when based on the restoration of muscle Se, buckwheat bran was 90% as high as Na selenite, but only 60% as high as SeMet. When using the ability of dietary Se to restore whole blood and liver glutathione peroxidase activity, buckwheat bran Se was 75-80% as high as Na selenite or SeMet. However, for the restoration of liver thioredoxin reductase, buckwheat bran Se was only 40% as high as Na selenite and 70% as high as SeMet. The relative bioavailability of Se from buckwheat bran with all variables considered was approximately 73% whether measured against Na selenite or SeMet. Although some variables indicated low bioavailability of Se from buckwheat bran, other factors such as Se speciation in the bran, digestibility of the bran, the cooking process, and combinations with other foods in the diet should be considered and analyzed before firm conclusions can be reached. PMID- 16251623 TI - Quantitative analysis of the benefits and risks of consuming farmed and wild salmon. AB - Contaminants in farmed Atlantic and wild Pacific salmon raise important questions about the competing health benefits and risks of fish consumption. A benefit-risk analysis was conducted to compare quantitatively the cancer and noncancer risks of exposure to organic contaminants in salmon with the (n-3) fatty acid associated health benefits of salmon consumption. Recommended levels of (n-3) fatty acid intake, as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), may be achieved by consuming farmed or wild salmon while maintaining an acceptable level of noncarcinogenic risk. However, the recommended level of EPA+DHA intake cannot be achieved solely from farmed or wild salmon while maintaining an acceptable level of carcinogenic risk. Although the benefit-risk ratio for carcinogens and noncarcinogens is significantly greater for wild Pacific salmon than for farmed Atlantic salmon as a group, the ratio for some subgroups of farmed salmon is on par with the ratio for wild salmon. This analysis suggests that risk of exposure to contaminants in farmed and wild salmon is partially offset by the fatty acid-associated health benefits. However, young children, women of child-bearing age, pregnant women, and nursing mothers not at significant risk for sudden cardiac death associated with CHD but concerned with health impairments such as reduction in IQ and other cognitive and behavioral effects, can minimize contaminant exposure by choosing the least contaminated wild salmon or by selecting other sources of (n-3) fatty acids. PMID- 16251624 TI - Nutritional status predicts primary subclasses of T cells and the lymphocyte proliferation response in healthy older women. AB - Aging is often associated with a dysregulation in immune function, particularly in T-cell responses, even in the healthy elderly. Adequate nutrition is important for optimal immune function. The literature on the relation of nutritional status with immune function in the elderly offers mixed findings. Because several nutrients can influence immune response, and there are interactions among nutrients, examining the association of various nutrients measured simultaneously with tests of immune function is important. We examined the association of protein, iron, zinc, vitamin B-12, and folic acid with tests of acquired immunity in healthy older women (76.7 +/- 7.0 y; n = 130). Discriminant analysis was used to identify the predictive subset of nutrients that could correctly classify subjects into the lowest or highest quartiles (< or =25th or >75th percentile) on various immune function tests (T cells and subsets and lymphocyte proliferation in response to culture with mitogens). Protein and iron status variables were identified in the predictive subset for all immune tests; in addition, zinc emerged in the predictive model for T cells and their subsets as well as for the proliferation response to concanavalin A. The probability of correctly classifying women into the lowest or highest quartiles of immune tests by the predictive subset of nutrition variables was high, i.e., 62.8-83.5% for T cells and their subsets, and 79.3-89.7% for the proliferation response to mitogens. In conclusion, protein, iron, and zinc were significant predictors of immune function in older women. Adequate status of these nutrients may help maintain immunity in older adults. PMID- 16251625 TI - Murine antigen-presenting cells are multifunctional in vitro biosensors for detecting the immunoactive potential of bovine milk products. AB - Antigen-presenting cells (APCs) are multifunctional components of the immune defense system. In this study, murine APCs were used as biosensors to detect immunologically active components of bovine milk and colostrum. By measuring changes in cell surface protein markers [major histocompatibility complex II, cluster designation (CD)40, CD86] and cytokines (tumor necrosis factor-alpha and interleukin-10) associated with APC activation, we identified a number of compounds that are immunoactive. The mouse macrophage cell line MH-S offered a simple and robust target for identification of immunoactives. The assay was shown to be adaptable for measuring immunoenhancing or immunosuppressive substances. Large-scale screening of milk extracts using this bioassay has the potential to identify substances that could be developed into nutraceuticals or pharmaceutical grade immunotherapeutics. PMID- 16251626 TI - The intestinal trophic response to enteral food is reduced in parenterally fed preterm pigs and is associated with more nitrergic neurons. AB - In term neonates, total parenteral nutrition (TPN) induces mucosal atrophy, whereas the first intake of milk is followed by intestinal growth. This may be explained in part by an NO-mediated increased blood flow. We hypothesized that the immature gut has an altered response to TPN and enteral nutrition. In Expt. 1, preterm caesarean-delivered pigs were administered elemental nutrients for 3 d, infused parenterally (TPN, n = 7) or enterally (TENT, n = 7). In Expt. 2, preterm pigs were fed sow's colostrum, cow's colostrum, or infant formula for 2 d after a 3-d TPN period (TPN-SOW, TPN-COW, TPN-FORM, n = 8-11). Intestinal morphology and the number of enteric neurons containing nitric oxide synthase-1 (NOS-1) were quantified. Both the TPN and TENT groups had increases in intestinal mass, circumference, and mucosal mass, volume, and surface density, relative to values at birth (+30-50%, P < 0.05). In Expt. 2, the magnitudes of the intestinal trophic responses to feeding were similar to those in Expt. 1, but were also associated with an increased number of nitrergic myenteric neurons and some mucosal damage, most frequently observed for the formula group. We conclude that 1) a short period of TPN does not induce mucosal atrophy in preterm pigs, whereas elemental nutrients infused luminally do not mimic the trophic response seen with milk diets, 2) enteral feeding of preterm pigs after a short period of TPN is associated with a modest, diet-dependent trophic response that may be related in part to the actions of an increased population of enteric NOS-1 neurons. PMID- 16251627 TI - Iron status and risk of cancers in the SU.VI.MAX cohort. AB - The aim of the present study was to evaluate the relation between iron status and cancer in a population of middle-aged adults living in France where iron supplementation and iron-fortified foods are rarely used. The SU.VI.MAX study is a randomized, double-blind, placebo-controlled primary prevention trial evaluating the effect of antioxidant supplementation on chronic diseases in women aged 35-60 and men aged 45-60 y. At baseline, concentrations of hemoglobin, serum transferrin and serum ferritin were measured in 10,197 subjects. Data on dietary intake were estimated from six 24-h dietary records completed during the first 2 study years and available for 5287 subjects. All cancer cases that occurred during the 7.5-y follow-up were validated. In men, baseline serum transferrin and serum ferritin concentrations did not differ between subjects with cancers (n = 467) and those without. In women, serum ferritin was higher (P < 0.0001) and serum transferrin tended to be lower (P < 0.08) in cancer cases. Iron status was not related to cancer risk in men, but women with serum ferritin concentrations > 160 microg/L had an increased risk of cancer (odds ratio = 1.88, 95% CI: 1.05,3.35). No relation was found between dietary iron intake and risk of all cancer sites combined for either men or women. Our results suggest that iron status is not a predictor of cancer risk in men, whereas a serum ferritin concentration > 160 microg/L may be associated with an increase in cancer risk in women. PMID- 16251628 TI - Vitamin a supplementation does not affect infants' immune responses to polio and tetanus vaccines. AB - It has been suggested that administering vitamin A with the measles vaccine may reduce the vaccine's immunogenicity. This trial examined the effect of supplementing vitamin A during the early months of life on infants' immune responses to tetanus and polio vaccines. Young infants (n = 1085) were enrolled and individually randomized into 1 of 4 groups in a factorial, double-blind, placebo-controlled trial. Three vitamin A supplementation strategies were investigated: 1) supplementation of breast-feeding mothers with 60 mg retinol equivalent (RE) vitamin A within 4 wk of delivery; 2) Expanded Program on Immunization (EPI)-linked supplementation of infants with 7.5 mg RE vitamin A at 6, 10, and 14 wk; and 3) combined mother and child supplementations. A 4th group in which mother and child were given placebos served as controls. Blood samples were collected from each child at 6 wk and 6 mo of age to measure antipolio antibody titer, antitetanus toxoid antibodies, and avidity of antibodies to tetanus. Of the infants randomized into the 4 arms of the study, 767 (71%) completed follow-up at 6 mo of age. Follow-up rates were similar in all 4 arms (69-72%, P = 0.8). Antibody titers were relatively high in all 4 groups at both 6 wk and 6 mo of age, with no differences among the groups. We found no evidence that vitamin A supplementation affects infants' antibody responses to tetanus toxoid or oral polio vaccine delivered at EPI contacts. PMID- 16251629 TI - The type of oil used for cooking is associated with the risk of nonfatal acute myocardial infarction in costa rica. AB - Palm oil and soybean oil are the 2 most widely used cooking oils in the world. Palm oil is consumed mainly in developing countries, where morbidity and mortality due to cardiovascular disease (CVD) are on the rise. Although claims about adverse or protective effects of these oils are commonly made, there are no epidemiologic studies assessing the association between these oils and cardiovascular disease endpoints. We examined whether consumption of palm oil relative to soybean oil and other unsaturated oils (predominantly sunflower) is associated with myocardial infarction (MI) in Costa Rica. The cases (n = 2111) were survivors of a first acute MI and were matched to randomly selected population controls (n = 2111). Dietary intake was assessed with a validated semiquantitative FFQ. Adipose tissue profiles of essential fatty acids were assessed to validate cooking oil intake and found to be consistent with self reported major oils used for cooking. The data were analyzed using conditional logistic regression. Palm oil users were more likely to have an MI than users of soybean oil [odds ratio (OR) = 1.33; 95% CI: 1.08-1.63] or other cooking oils (OR = 1.23; CI: 0.99-1.52), but they did not differ from users of soybean oil with a high trans-fatty acid content (OR = 1.14; CI: 0.84-1.56). These data suggest that as currently used in Costa Rica, and most likely in many other developing countries, the replacement of palm oil with a polyunsaturated nonhydrogenated vegetable oil would reduce the risk of MI. PMID- 16251630 TI - Polymorphisms in the CYP19 gene may affect the positive correlations between serum and urine phytoestrogen metabolites and plasma androgen concentrations in men. AB - Phytoestrogens have been hypothesized to protect against prostate cancer via modulation of circulating androgen concentrations. We conducted a cross-sectional study of 267 men in the Norfolk arm of the European Prospective Investigation into Cancer and Nutrition (EPIC) cohort with 2 aims: first, to investigate the association between phytoestrogen exposure (measured from diet, urine, and serum) and plasma concentrations of sex hormone-binding globulin (SHBG), androstanediol glucuronide, testosterone and Free Androgen Index (FAI); and second, whether the association may be modified by polymorphisms in CYP19 and SHBG genes. Dietary daidzein and genistein intakes were obtained from food diaries and computed using an in-house food composition database. Urinary and serum concentrations of 3 isoflavones (daidzein, genistein, glycitein), 2 daidzein metabolites O desmethylangolensin (O-DMA) and 2 lignan metabolites (enterodiol and enterolactone) were measured using mass spectrometry. There was no association between dietary, urinary, and serum phytoestrogens and plasma SHBG concentrations. Enterolactone was positively associated with plasma androstanediol glucuronide concentrations (urinary enterolactone: r = 0.127, P = 0.043; serum enterolactone: r = 0.172, P = 0.006) and FAI (urinary enterolactone: r = 0.115, P = 0.067; serum enterolactone: r = 0.158, P = 0.011). Both urinary and serum equol were associated with plasma testosterone (urinary equol: r = 0.332, P = 0.013; serum equol: r = 0.318, P = 0.018) and FAI (urinary equol: r = 0.297, P = 0.027; serum equol: r = 0.380, P = 0.004) among men with the TT genotype but not the CC or CT genotypes (r = -0.029 to -0.134, P = 0.091-0.717) for the CYP19 3'untranslated region (UTR) T-C polymorphism. Urinary and serum enterolactone showed similar genotype-dependent associations with testosterone but not with FAI. In this first study on phytoestrogen-gene associations in men, we conclude that enterolactone and equol are positively associated with plasma androgen concentrations, and interactions with CYP19 gene may be involved. PMID- 16251631 TI - Daidzein-sulfate metabolites affect transcriptional and antiproliferative activities of estrogen receptor-beta in cultured human cancer cells. AB - Daidzein (D), a soy isoflavone, is almost completely metabolized in the gut and liver. This biotransformation converts D to more water-soluble products and may affect its biological activity. The ability of daidzein metabolites to modulate 17beta-estradiol (E2)-sensitive gene transcription, cell growth, and a proapoptotic cascade was determined in human cancer cells devoid of any estrogen receptor (ER) and rendered E2 sensitive after transfection with ERbeta. The data show that D and some but not all of its metabolites 1) induce promoter activity, 2) reduce proliferation, 3) promote p38/mitogen-activated protein kinase (MAPK) phosphorylation, and 4) activate a proapoptotic cascade involving the cleavage of caspase-3 and its substrate poly(ADP-ribose)polymerase (PARP) in human cancer cells in an ERbeta-dependent manner. Pretreatment of cells with ICI 182,780, a pure antiestrogen, completely prevented the actions of D and its metabolites. These findings highlight the important and complex influence of metabolic transformation on key physiological effects of isoflavones and demonstrate the need to take biotransformation into account when assessing the potential health benefits of consuming soy isoflavones. PMID- 16251632 TI - Nutritional "omics" technologies for elucidating the role(s) of bioactive food components in colon cancer prevention. AB - Evidence continues to implicate dietary components and genetic susceptibilities as important determinants of cancer risk and tumor behavior. Variation in cancer incidence among and within populations with similar dietary patterns suggests that an individual's response may reflect interactions with genetic factors, which may modify gene, protein, and metabolite expression patterns. Nutrigenomics, defined as the interaction between nutrition and an individual's genome, will likely provide important clues about responders and nonresponders. In this symposium, the role of bioactive food components in colon cancer susceptibility was used to exemplify the application of "omic" technologies for cancer prevention. Topics that were addressed included dietary changes and gene polymorphisms (nutrigenetics), DNA methylation (nutritional epigenomics), gene expression (nutritional transcriptomics), altered formation or bioactivation of proteins (proteomics), and characterizing how the quantity and timing of exposure influence small molecular weight cellular constituents (metabolomics). The final presentation focused on exfoliated cells as a surrogate sample for the evaluation of bioactive food components in cancer prevention. The goal of the symposium was to provide an example of each of the "omic" technologies as they relate to nutrition, cancer risk, and tumor behavior, and to help the participants understand that an integrated framework that simultaneously examines all of the "omic" technologies is needed. PMID- 16251633 TI - Nutrigenetics in cancer research--folate metabolism and colorectal cancer. AB - The B vitamin folate is essential for one-carbon transfer reactions, including those related to the methylation of DNA or other substrates and nucleotide synthesis. Epidemiologic and experimental studies implicate low-folate intakes in elevated risk of colorectal neoplasia and suggest that biologic mechanisms underlying this relation include disturbances in DNA methylation patterns or adverse effects on DNA synthesis and repair. With the completion of the Human Genome Project, a vast amount of data on inherited genetic variability has become available. This genetic information can be used in studies of molecular epidemiology to provide information on multiple aspects of folate metabolism. First, studies linking polymorphisms in folate metabolism to an altered risk of cancer provide evidence for a causal link between this pathway and colorectal carcinogenesis. Second, studies on genetic characteristics can help clarify whether certain individuals may benefit from higher or lower intakes of folate or nutrients relevant to folate metabolism. Third, studies on genetic polymorphisms can generate hypotheses regarding possible biologic mechanisms that connect this pathway to carcinogenesis. Last, genetic variability in folate metabolism may predict survival after a cancer diagnosis, possibly via pharmacogenetic effects. To solve the puzzle of the folate-cancer relation, a transdisciplinary approach is needed that integrates knowledge from epidemiology, clinical studies, experimental nutrition, and mathematical modeling. This review illustrates knowledge that can be gained from molecular epidemiology in the context of nutrigenetics, and the questions that this approach can answer or raise. PMID- 16251634 TI - Nutritional epigenetics: impact of folate deficiency on DNA methylation and colon cancer susceptibility. AB - The inheritance of information based on gene expression levels is known as epigenetics, as opposed to genetics, which refers to information transmitted on the basis of gene sequence. In contrast to genetic changes observed in cancer, epigenetic changes are gradual in onset and are progressive, their effects are dose-dependent and are potentially reversible. These observations present new opportunities in cancer-risk modification and prevention using dietary and lifestyle factors and potential chemopreventive drugs. In this regard, folate, a water-soluble B vitamin, has been a focus of intense interest because of an inverse association between folate status and the risk of several malignancies (in particular, colorectal cancer) and of its potential ability to modulate DNA methylation. DNA methylation is an important epigenetic determinant in gene expression, in the maintenance of DNA integrity and stability, in chromosomal modifications, and in the development of mutations. Aberrant patterns and dysregulation of DNA methylation are mechanistically related to colorectal carcinogenesis. Folate plays an essential role in one-carbon transfer involving re-methylation of homocysteine to methionine, thereby ensuring the provision of S adenosylmethionine, the primary methyl group donor for most biological methylation reactions. The portfolio of evidence from animal, human, and in vitro studies suggests that the effects of folate deficiency and supplementation on DNA methylation are gene and site specific, and appear to depend on cell type, target organ, stage of transformation, and the degree and duration of folate depletion. PMID- 16251635 TI - Dietary components modify gene expression: implications for carcinogenesis. AB - Mouse genetic models that probe important pathways in intestinal cell maturation, such as cell-cycle regulation, apoptosis, and, especially, lineage specific differentiation, have provided profound insight into the underlying mechanisms of intestinal tumor formation and progression. However, a wealth of epidemiological and experimental data indicates that environment, especially the diet, is a principal determinant of relative risk for tumor development. We have demonstrated that even in mouse models in which tumor incidence is strongly initiated by genetic manipulation of genes, such as Apc, p21(WAF1/cip1), and p27(Kip1), a Western-style diet that is high in fat and low in calcium and vitamin D can dramatically increase and accelerate tumor formation. Moreover, experiments show that modulation of calcium and vitamin D levels can substantially influence tumor formation in both the mouse genetic models, as well as in a new dietary model that appears to mimic the development of sporadic colon cancer. Finally, analysis of gene expression profiles provides important insights into how diets may alter metabolic profiles and regulatory pathways that influence probability of tumor formation in the histologically and physiologically normal intestinal mucosa. PMID- 16251636 TI - New nutrition, proteomics, and how both can enhance studies in cancer prevention and therapy. AB - The increased application of MS technologies to nutrition and cancer prevention research has enabled unique insights into the health benefits of polyphenols. Polyphenols are phytochemicals that appear to have chemical properties that provide valuable health benefits when ingested. In particular, experiments have suggested that grape seed proanthocyanidins, oligomers of the catechin family of polyphenols, may have health benefits, possibly due to their capacity to be oxidized. Two-dimensional gel proteomics technology identified specific rat brain proteins that were differentially affected after ingestion of grape seed extract. Beneficial changes in the expression of these proteins were observed relative to changes seen in the brains of Alzheimer disease patients at autopsy and of transgenic mouse models of dementia. These findings were consistent with the hypothesis that grape seed polyphenols may have neuroprotective activity. Previous experiments showed that grape seed extract was significantly chemopreventive in a rat model of breast cancer, but this depended on the specific diet in which the grape seed was administered. Thus, phytochemicals such as polyphenols may have health benefits in mammalian tissues unrelated to classical nutritional deficiency models. This report illustrates how experimental approaches that combine proteomics technologies with a dietary intervention with specific phytochemicals in normal animals can enhance studies on cancer prevention and treatment. PMID- 16251637 TI - Exfoliated colonic epithelial cells: surrogate targets for evaluation of bioactive food components in cancer prevention. AB - There is significant evidence supporting the hypotheses that lifestyle, diet, and bioactive components in foods are important modifiers of cancer risk. However, our ability to assess host response noninvasively is limited. To overcome this, we have developed a technology to isolate several million viable exfoliated somatic colonic cells from a small sample of stool (0.5-1.0 g) by a procedure known as somatic cell sampling and recovery (SCSR). Orally administered carotenoids appear in these cells several days after consuming the supplement, usually showing a peak concentration between 5-7 d after their ingestion. The time lag observed for the appearance of orally administered carotenoids in SCSR cells corresponds to the turnover rate of the colonic mucosa. This is an example of how changes in cell turnover rates can be carefully assessed using the SCSR system. The specific mechanisms by which individual constituents of diet affect the cancer process are not fully understood. However, host response to dietary constituents may be investigated, noninvasively, by following the modulation of tumor-associated molecular markers in these exfoliated SCSR cells. We have demonstrated the feasibility of using SCSR cells to detect the expression of carcinoembryonic antigen, CD44, and its splice variants, c-myc, c-erbb2, and mutations in the p53 gene. In this regard, SCSR cells are a readily available surrogate cellular target that may serve to monitor changes in cell turnover, differentiation, and expression of cancer-associated biomarkers that are likely to be modulated by bioactive food components. PMID- 16251638 TI - The influence of vitamin D on bone health across the life cycle. PMID- 16251639 TI - Developmental origins of osteoporotic fracture: the role of maternal vitamin D insufficiency. AB - Osteoporosis is a major cause of morbidity and mortality through its association with age-related fractures. Although most efforts in fracture prevention have been directed at retarding the rate of age-related bone loss and reducing the frequency and the severity of trauma among elderly people, evidence is growing that peak bone mass is an important contributor to bone strength during later life. The normal patterns of skeletal growth have been well characterized in cross-sectional and longitudinal studies. It has been confirmed that boys have higher bone-mineral content, but not volumetric bone density, than girls. Furthermore, there is a disassociation between the peak velocities for height gain and bone mineral accrual in both genders. Puberty is the period during which volumetric density appears to increase in both axial and appendicular sites. Many factors influence the accumulation of bone mineral during childhood and adolescence, including heredity, gender, diet, physical activity, endocrine status, and sporadic risk factors (e.g., cigarette smoking). In addition to these modifiable factors during childhood, evidence has also accrued that fracture risk might be programmed during intrauterine life. Epidemiological studies have demonstrated a relationship between birthweight, weight in infancy, and adult bone mass. This appears to be mediated through modulation of the set-point for basal activity of endocrine systems such as the GH/IGF-1 and parathyroid hormone/vitamin D axes. Maternal vitamin D insufficiency is associated with reduced bone mineral acquisition during intrauterine and early postnatal life. Furthermore, both low birth size and poor childhood growth are directly linked to the later risk of hip fracture. The optimization of maternal nutrition and intrauterine growth should also be included within preventive strategies against osteoporotic fracture, albeit for future generations. PMID- 16251640 TI - Vitamin D, parathyroid hormone, and bone mass in adolescents. AB - This article provides a review of the evidence identifying the factors related to vitamin D status in adolescents. The prevalence of vitamin D deficiency based on 25-hydroxyvitamin D [25(OH)D] of <25 nmol/L ranges from 0 to 32% depending on the season measured and the latitude of the population assessed. The factors that have been reported to affect serum 25(OH)D in adolescents include ethnicity, gender, puberty stage, parathyroid hormone (PTH), dietary vitamin D intake, and sun exposure. Vitamin D supplementation studies are limited to small populations and with supplementation focused on winter months when sunlight may be inadequate. The effects of vitamin D status and supplementation on bone assessment provide varied results. Differences in study design, modalities of bone assessment, and stage of puberty could contribute to disparate findings. Overall, the results from the available literature provide more questions than answers concerning the role of vitamin D in bone accrual in adolescents. PMID- 16251641 TI - The vitamin D epidemic and its health consequences. AB - Vitamin D deficiency is now recognized as an epidemic in the United States. The major source of vitamin D for both children and adults is from sensible sun exposure. In the absence of sun exposure 1000 IU of cholecalciferol is required daily for both children and adults. Vitamin D deficiency causes poor mineralization of the collagen matrix in young children's bones leading to growth retardation and bone deformities known as rickets. In adults, vitamin D deficiency induces secondary hyperparathyroidism, which causes a loss of matrix and minerals, thus increasing the risk of osteoporosis and fractures. In addition, the poor mineralization of newly laid down bone matrix in adult bone results in the painful bone disease of osteomalacia. Vitamin D deficiency causes muscle weakness, increasing the risk of falling and fractures. Vitamin D deficiency also has other serious consequences on overall health and well-being. There is mounting scientific evidence that implicates vitamin D deficiency with an increased risk of type I diabetes, multiple sclerosis, rheumatoid arthritis, hypertension, cardiovascular heart disease, and many common deadly cancers. Vigilance of one's vitamin D status by the yearly measurement of 25 hydroxyvitamin D should be part of an annual physical examination. PMID- 16251642 TI - 13th EUPHA (European Public Health Association) Conference, Graz, Austria, 10-12 November 2005. Abstracts. PMID- 16251720 TI - Cell surface-expressed moesin-like HDL/apoA-I binding protein promotes cholesterol efflux from human macrophages. AB - HDL and its major component, apolipoprotein A-I (apoA-I), play a central role in reverse cholesterol transport. We recently reported the involvement of a glycosylphosphatidylinositol anchor (GPI anchor) in the binding of HDL and apoA-I on human macrophages, and purified an 80 kDa HDL/apoA-I binding protein. In the present study, we characterized the GPI-anchored HDL/apoA-I binding protein from macrophages. The HDL/apoA-I binding protein was purified from macrophages and digested with endopeptidase, and the resultant fragments were sequenced. Cholesterol efflux, flow cytometry, immunoblotting, and immunohistochemical analyses were performed to characterize the HDL/apoA-I binding protein. Two parts of seven amino acid sequences completely matched those of moesin. Flow cytometry, immunoblotting, and immunohistochemistry using anti-moesin antibody showed that the HDL/apoA-I binding protein was N-glycosylated and expressed on the cell surface. It was termed moesin-like protein. Treatment of macrophages with anti moesin antibody blocked the binding of HDL/apoA-I and suppressed cholesterol efflux. The moesin-like protein was exclusively expressed on macrophages and was upregulated by cholesterol loading and cell differentiation. Our results indicate that the moesin-like HDL/apoA-I binding protein is specifically expressed on the surface of human macrophages and promotes cholesterol efflux from macrophages. Matsuyama, A, N. Sakai, H. Hiraoka, K-i. Hirano, and S. Yamashita. Cell surface expressed moesin-like HDL/apoA-I binding protein promotes cholesterol efflux from human macrophages. PMID- 16251723 TI - Caught with their proboscises in the sugar water. PMID- 16251721 TI - FXR regulates organic solute transporters alpha and beta in the adrenal gland, kidney, and intestine. AB - Expression of the farnesoid X receptor (FXR; NR1H4) is limited to the liver, intestine, kidney, and adrenal gland. However, the role of FXR in the latter two organs is unknown. In the current study, we performed microarray analysis using RNA from H295R cells infected with constitutively active FXR. Several putative FXR target genes were identified, including the organic solute transporters alpha and beta (OSTalpha and OSTbeta). Electromobility shift assays and promoter reporter studies identified functional farnesoid X receptor response elements (FXREs) in the promoters of both human genes. These FXREs are conserved in both mouse genes. Treatment of wild-type mice with 3-(2,6-dichlorophenyl)-4-(3' carboxy-2-chloro-stilben-4-yl)-oxymethyl-5-isopropyl-isoxazole (GW4064), a synthetic FXR agonist, induced OSTalpha and OSTbeta mRNAs in the intestine and kidney. Both mRNAs were also induced when wild-type, but not FXR-deficient (FXR-/ ), adrenals were cultured in the presence of GW4064. OSTalpha and OSTbeta mRNA levels were also induced in the adrenals and kidneys of wild-type, but not FXR-/ , mice after the increase of plasma bile acids in response to the hepatotoxin alpha-naphthylisothiocyanate. Finally, overexpression of human OSTalpha and OSTbeta facilitated the uptake of conjugated chenodeoxycholate and the activation of FXR target genes. These results demonstrate that OSTalpha and OSTbeta are novel FXR target genes that are expressed in the adrenal gland, kidney, and intestine. PMID- 16251724 TI - Trapped inside. PMID- 16251722 TI - Absorption and lipoprotein transport of sphingomyelin. AB - Dietary sphingomyelin (SM) is hydrolyzed by intestinal alkaline sphingomyelinase and neutral ceramidase to sphingosine, which is absorbed and converted to palmitic acid and acylated into chylomicron triglycerides (TGs). SM digestion is slow and is affected by luminal factors such as bile salt, cholesterol, and other lipids. In the gut, SM and its metabolites may influence TG hydrolysis, cholesterol absorption, lipoprotein formation, and mucosal growth. SM accounts for approximately 20% of the phospholipids in human plasma lipoproteins, of which two-thirds are in LDL and VLDL. It is secreted in chylomicrons and VLDL and transferred into HDL via the ABCA1 transporter. Plasma SM increases after periods of large lipid loads, during suckling, and in type II hypercholesterolemia, cholesterol-fed animals, and apolipoprotein E-deficient mice. SM is thus an important amphiphilic component when plasma lipoprotein pools expand in response to large lipid loads or metabolic abnormalities. It inhibits lipoprotein lipase and LCAT as well as the interaction of lipoproteins with receptors and counteracts LDL oxidation. The turnover of plasma SM is greater than can be accounted for by the turnover of LDL and HDL particles. Some SM must be degraded via receptor-mediated catabolism of chylomicron and VLDL remnants and by scavenger receptor class B type I receptor-mediated transfer into cells. PMID- 16251725 TI - Neurochemical insights. AB - The 20th biennial meeting of the International Society for Neurochemistry was recently held in Innsbruck, Austria. This meeting gave an overview of the latest findings in the field of molecular mechanisms and diagnosis of neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis, and prion disease. There was a focus on the molecular pathogenesis of protein misfolding in these disorders as well as on the association between oxidative metabolism and neurological diseases. RNA interference, metal chelators, and the use of metallopeptidases were discussed as possible therapeutic strategies. PMID- 16251726 TI - Behavioral and neurophysiological analyses of dynamic learning processes. AB - In this article, the authors address two topics relevant to the study of the brain basis of associative learning. In Part 1, they compare and contrast the patterns and time course of dynamic learning-related neural activity that have been reported in the medial temporal lobe, premotor cortex, prefrontal cortex, and striatum during various associative learning tasks. In Part 2, they examine the statistical methodologies that have been used to analyze both behavioral learning and learning-related neural activity. They describe a state-space model of behavioral learning that provides accurate estimates of dynamic learning processes and a point-process filter algorithm that tracks the dynamic changes in neural activity on a millisecond time scale. Future challenges for these statistical methodologies and their application to the study of the brain basis of associative learning are discussed. PMID- 16251727 TI - Transformations of visuospatial images. AB - Transformations of visuospatial mental images are important for action, navigation, and reasoning. They depend on representations in multiple spatial reference frames, implemented in the posterior parietal cortex and other brain regions. The multiple systems framework proposes that different transformations can be distinguished in terms of which spatial reference frame is updated. In an object-based transformation, the reference frame of an object moves relative to those of the observer and the environment. In a perspective transformation, the observer's egocentric reference frame moves relative to those of the environment and of salient objects. These two types of spatial reference frame updating rely on distinct neural processing resources in the parietal, occipital, and temporal cortex. They are characterized by different behavioral patterns and unique individual differences. Both object-based transformations and perspective transformations interact with posterior frontal cortical regions subserving the simulation of body movements. These interactions indicate that multiple systems coordinate to support everyday spatial problem solving. PMID- 16251728 TI - Current models and future directions for understanding the neural circuitries of maternal behaviors in rodents. AB - Maternal behaviors in rodents include a number of subcomponents, such as nursing, nest building, licking and grooming of pups, pup retrieval, and maternal aggression. Because each behavior involves a unique motor pattern, a unique ensemble neural circuitry must underlie each behavior. To what extent there is overlap in terms of brain regions and specific neurons for each circuit is being actively investigated. This review will first examine overlapping and separate components of pup retrieval and maternal aggression circuitries while examining a central role for medial preoptic area (MPA) in both behaviors. With an emphasis on experimental approaches, the review will then highlight recent findings and propose future directions for understanding maternal behavior regulation. Finally, examples for why studying the neural basis of maternal behaviors can bring insights to other areas of neuroscience, such as feeding, addiction, and anxiety and aggression regulation will be provided. PMID- 16251731 TI - Methodologies to study implantation in mice. AB - Pregnancy begins with fertilization of the ovulated oocyte by the sperm. After fertilization, the egg undergoes time-dependent mitotic division while trying to reach the blastocyst stage and the uterus for implantation. Uterine preparation for implantation is regulated by coordinated secretions and functions of ovarian sex steroids. The first sign of contact between the blastocyst and the uterus can be detected experimentally by an intravenous blue dye injection as early as the end of day 4 or the beginning of day 5 of pregnancy. This blastocyst-uterine attachment reaction leads to stromal decidual reaction only at sites of implantation. The process of implantation can be postponed and reinstated experimentally by manipulating ovarian estrogen secretion. Stromal decidualization can also be induced experimentally in the hormonally prepared uterus in response to stimuli other than the embryo. Fundamental biological questions surrounding these essential features of early pregnancy can be addressed through the application of various techniques and manipulation of this period of early pregnancy. This chapter describes the routine laboratory methodologies to study the events of early pregnancy, with special emphasis on the implantation process in mice. PMID- 16251732 TI - Blastocyst culture. AB - Experimental models of blastocyst development based on in vitro culture have played a prominent role in advancing our understanding of peri-implantation development, a process that is relatively inaccessible in vivo. Blastocyst culture provides a robust approach for examining embryonic interactions with the microenvironment under highly controlled conditions. Major events that occur in utero can be followed in vitro, including blastocyst expansion, hatching, and adhesion to extracellular matrices. This chapter will describe a method for obtaining and culturing mouse blastocysts. Morphological changes that occur during blastocyst culture will be discussed and related to the corresponding development in utero. Finally, quantitative assays will be detailed for monitoring peri-implanatation development of the trophoblast in vitro. PMID- 16251733 TI - Isolation of hormone responsive uterine stromal cells: an in vitro model for stromal cell proliferation and differentiation. AB - The female sex hormones estrogen and progesterone stimulate proliferation and differentiation of human and rodent uterine cells. The purpose of this chapter is to provide a method for isolating hormone-responsive rat uterine stromal cell lines that can be used to study steroid control of the cell cycle. Uteri from ovariectomized rats are differentially digested with trypsin to separate epithelial and stromal cells. The stromal cells are cultured in a standard growth medium containing 10% fetal bovine serum. After several passages, the purity of the stromal cell lines is determined using immunocytochemistry. Cell proliferation is studied by culturing the stromal cells in serum-free medium containing sex steroids and other mitogens. Cell cycle progression is assessed by flow cytometry, 3H-thymidine and BrdU incorporation, whereas proliferation is monitored using the MTT assay. Cell cycle regulators are visualized by Northern and Western blotting whereas cyclin-cyclin-dependent kinase activity is monitored using immune complex kinase assays. Uterine stromal cell lines isolated using the methods reported in this chapter provide a suitable model system to investigate the signal transduction events that stimulate hormone-dependent control of the cell cycle. PMID- 16251734 TI - Rat decidual cell cultures. AB - Pregnancy requires profound reorganization of the different tissues forming the uterus. Growth and differentiation of the uterine endometrial cells give rise to the decidual tissue, a transitory organ, which plays a key role in fetal survival. In this chapter, we describe a technique for the dispersion and the separation of the two different decidual cell subpopulations with high yield and viability. We also detail a cell culture method, which allows the maintenance of the function and life span of these highly purified decidual cells when cultured either separately or in a co-culture system. PMID- 16251735 TI - The immortalization of human endometrial cells. AB - The loss of replicative potential with each cell division has been attributed to the progressive shortening of telomeres. This "mitotic clock" occurs because most normal human cells are telomerase-negative. Telomerase is a multicomponent enzyme that prevents loss of telomeric DNA associated with normal cell division. Transfection of cells with vectors expressing the catalytic subunit of human telomerase (hTERT) is often sufficient for immortalization. In this article, we will address this approach in the establishment of immortalized endometrial cells and its value in facilitating in vitro studies. PMID- 16251736 TI - Sheep uterine gland knockout (UGKO) model. AB - Endometrial gland development is a postnatal event in the ovine uterus that can be inhibited epigenetically by chronic exposure of ewe lambs to a synthetic progestin after birth. The uterus of neonatally progestinized ewes lack endometrial glands and display a uterine gland knockout (UGKO) phenotype. Progestin ablation of endometrial gland development is specific, because it does not affect development of extra-uterine reproductive tract structures or the hypothalamic-pituitary-ovarian axis. The UGKO ewe is a useful model for study of uterine development and the role of endometrial glands in uterine function during the estrous cycle and pregnancy. UGKO ewes exhibit altered estrous cycles due to the inability of the uterus to produce luteolytic pulses of prostaglandin F2alpha. UGKO ewes are infertile, and blastocysts hatch normally but fail to survive or elongate during early pregnancy. This pregnancy defect is primarily due to the absence of endometrial glands and their secretions rather than alterations in expression of either anti-adhesive or adhesive molecules on the endometrial epithelium. Genomics and proteomics are being used to identify specific components of histotroph that are absent or diminished in the UGKO ewe and will serve as markers of endometrial function and uterine receptivity. PMID- 16251737 TI - A baboon model for inducing endometriosis. AB - Endometriosis is a disease that is associated with severe pelvic pain and is a major cause of infertility in women. It is an enigmatic disease whose etiology and pathophysiology has been studied to a limited extent. The events associated with the establishment of the disease and mechanisms associated with infertility are difficult to assess in a systemic manner in women. In order to understand the early and progressive events associated with the establishment of the disease, we have developed a baboon model in which the disease can be induced. This induction manifests itself in a manner that recapitulates the spontaneous disease. The advantage of the induced model is that the progressive changes in both the ectopic and eutopic endometrium can be studied in a nonhuman primate model at specific times during the menstrual cycle and as the disease process continues. PMID- 16251738 TI - A baboon model for simulating pregnancy. AB - Estrogen and progesterone secreted by the corpus luteum regulate the function of the uterine endometrium in preparation for pregnancy. Embryonic signals superimposed on this steroid hormone-primed uterus further modulate the uterine environment to make it conducive to pregnancy. Understanding the signaling mechanisms that initiate the embryonic-maternal dialog in humans is not feasible. In an effort to elucidate the role of chorionic gonadotropin as a mediator of endometrial function in addition to its luteotrophic role, we have developed a simulated pregnant model in the baboon. Infusion of chorionic gonadotropin in a manner that mimics blastocyst transit induces major changes in the morphology and secretory activity of the endometrium. This model provides a method by which the function of various embryonic factors on endometrial can be tested in an in vivo model. PMID- 16251739 TI - The common marmoset monkey as a model for implantation and early pregnancy research. AB - This chapter describes methods used to investigate implantation in the common marmoset monkey, Callithrix jacchus. A reverse-transcriptase polymerase chain reaction-strategy with which to detect transcripts for steroid receptors and enzymes involved in estradiol biosynthesis is described, and an immunohistochemistry approach for detecting proteins within the implantation site is presented. PMID- 16251740 TI - Mouse trophoblast stem cells. AB - The trophectoderm is one of the earliest cell types to differentiate in the forming mammalian embryo. It is responsible for the initial implantation and the formation of the trophoblast components of the placenta, an organ essential for nutrient and waste exchange between the fetus and its mother. The trophoblast can be modeled in vitro using trophoblast stem cells. Trophoblast stem cells require fibroblast growth factor (FGF)4, heparin, and contact with embryonic fibroblasts, or fibroblast-conditioned medium. They grow as tight epithelial colonies, which express markers of the early trophectoderm and have been shown to contribute to all of the components of the placenta through chimera studies. These cells can be passaged indefinitely and can be differentiated by removal of FGF4 and fibroblasts and will express genetic markers of later placental cell types. This chapter will discuss the initial derivation of trophoblast stem cells from the blastocyst stage, maintenance, differentiation, flow cytometry and transfection techniques that can be used with these cells. PMID- 16251742 TI - Rcho-1 trophoblast stem cells: a model system for studying trophoblast cell differentiation. AB - The biology of trophoblast cell development can be investigated using in vitro model systems. The Rcho-1 trophoblast stem cell line was derived from a rat choriocarcinoma and is an effective tool for elucidating regulatory mechanisms controlling trophoblast cell differentiation. In this chapter, we describe methods used in the maintenance and manipulation of the Rcho-1 trophoblast cell line. PMID- 16251741 TI - Connexins and trophoblast cell lineage development. AB - The mouse is a valuable model for studying basic mechanisms of gene regulation in trophoblast cell lineage differentiation. Elements of placental development are conserved across species, including trophoblast proliferation, differentiation, migration, and vessel invasion. Among the regulatory processes, direct intercellular communication between trophoblast cells via gap junction channels seems to play a crucial role in placental development and physiology. Here we describe in detail the generation of trophoblast stem (TS) cell lines from connexin-deficient mice. The design of differentiation and proliferation assays are specified including marker gene sets which are important for analyzing and comparing the differentiation capacity of the connexin-deficient TS cell lines. Furthermore, we show that TS cells are capable of forming tumors after subcutaneous injection into nude mice, providing the opportunity to investigate trophoblast invasion into host vessels in vivo. PMID- 16251743 TI - Bovine trophoblast cell culture systems: a technique to culture bovine trophoblast cells without feeder cells. AB - Bovine trophoblastic cells are the first cells to differentiate during embryogenesis and play pivotal role in morphological and physiological development of the placenta. We have developed culture systems for bovine trophoblast stem cells isolated from in vitro fertilized blastocysts in the absence of feeder cells. These cells continuously proliferate in Dulbecco's modified Eagle's/F12 culture medium supplemented with bovine endometrial fibroblast-conditioned medium. The cells possess epithelial morphology, express cytokeratin, and form dome-like structures (vesicles). Methods for the maintenance, subculture, storage, and measurement of bovine trophoblast stem cell growth are described. The cells exhibit characteristics of bovine trophoblastic stem cells and possess the ability to differentiate into binucleate cells and express placental lactogen, prolactin-related protein-1, pregnancy-associated glycoprotein-1, and interferon tau. PMID- 16251744 TI - In vitro induction of trophoblast from human embryonic stem cells. AB - Human embryonic stem (ES) cells can proliferate without a known limit and can form advanced derivatives of all three embryonic germ layers. What is less widely appreciated is that human ES cells can also form the extra-embryonic tissues that differentiate from the embryo before gastrulation. The use of human ES cells to derive early human trophoblast is particularly valuable, because it is difficult to obtain from other sources and is significantly different from mouse trophoblast. Here we describe a method by using bone morphogenetic protein (BMP)4, a member of the transforming growth factor (TGF)-beta superfamily, to induce the differentiation of human ES cells to trophoblast. Immunoassays (as well as DNA microarray and reverse-transcription polymerase chain reaction analyses--data not shown) demonstrate that the differentiated cells express a range of trophoblast markers and secrete placental hormones. When plated at low density, the BMP4-treated cells form syncytia that express chorionic gonadotrophin (CG). This technique underscores fundamental differences between human and mouse ES cells, which differentiate poorly, if at all, to trophoblast. Human ES cells thus provide a tool for studying the differentiation and function of early human trophoblast and could provide a new understanding of some of the earliest differentiation events of human postimplantation development. PMID- 16251745 TI - Isolation and culture of term human trophoblast cells. AB - Experimentation with most human cell types is restricted to the use of cell lines, and this limits our ability to extrapolate interpretations to the in vivo condition. However, in studying human trophoblast cells, we have a unique opportunity to obtain large quantities of readily available human tissue. In this chapter, we outline the methodology for purification of human trophoblast cells from term placentas. The procedures are based on enzymatic dissociation of villous placental tissue, followed by gradient centrifugation and immunomagnetic bead purification. Purity may be assessed by immunocytochemistry or flow cytometry using a number of markers to identify both cytotrophoblast cells and cellular contaminants. The resulting cytotrophoblast cell populations have excellent viability and purity, and may be subjected to long-term culture. PMID- 16251746 TI - Production of human trophoblast cell lines. AB - Our understanding of important biological phenomena such as invasion, migration, and apoptosis has advanced greatly through the prudent use of in vitro models based on isolated cells in culture. Established cell lines are readily manipulated using simple molecular biological techniques and their abundance as homogenous populations allows the rapid accumulation of statistically significant data. The study of human trophoblast function in vitro has been hampered by the difficulties associated with obtaining and culturing primary human trophoblasts including the paucity of material and contamination with other cell types. This chapter describes a cheap and simple method for the production of human trophoblast cell lines using poly-L-ornithine. It details the production, isolation and initial characterization of these cells and provides simple tips on how to store and maintain a bank of cells for future needs. PMID- 16251747 TI - Culture and transfection of human choriocarcinoma cells. AB - In vitro models for human trophoblasts were initially established more than three decades ago from isolated choriocarcinomas. They have proven to be extremely valuable for the study of the cellular, molecular, and endocrine aspects of human trophoblasts. This chapter describes basic methods for culture and maintenance of the Jeg-3, Jar, and BeWo human choriocarcinoma cell lines as well as an effective paradigm for introducing DNA into the cells. PMID- 16251748 TI - In vitro methods for studying vascularization of the murine allantois and allantoic union with the chorion. AB - Despite the importance of the definitive chorio-allantoic placenta in fetal survival, fetal development, and long-term health of the adult, little is known about how the placenta's individual components, the allantois and the chorion, proliferate and develop. In this chapter, two techniques will be described: (1) explanting murine allantoises for culture in isolation, and (2) grafting murine allantoises into living whole mouse embryos. Together, these will enable study of differentiation of allantoic mesoderm into the umbilical vasculature, and the mechanism(s) by which the allantois unites with the chorion to form the chorio allantoic placenta. PMID- 16251749 TI - Phenotypic analysis of the mouse placenta. AB - Placental development is a dynamic and complex process and much of our current understanding of the underlying molecular processes comes from analysis of targeted gene mutations in mice. There are more than 50 strains of mutant mice that have placental defects, and it has become widely appreciated that placental defects should be suspected in cases where embryonic lethality is observed. The degree to which these phenotypes are investigated is highly variable, owing to a general lack of expertise in the field. However, there has been considerable progress in developing techniques and reagents for analyzing placental phenotypes that are relatively simple to apply and that should be accessible to all investigators. This chapter provides a basic outline of the strategies for the general identification and then the subsequent detailed investigation of placental phenotypes. PMID- 16251751 TI - Analysis of the structure of the ruminant placenta: methods of fixation, embedding, and antibody localization at light and electron microscope levels. PMID- 16251750 TI - Phenotypic analysis of the rat placenta. AB - The rat is an important model for studying the biology of trophoblast-uterine development. This chapter describes methods that are useful for the characterization of the rat uteroplacental compartment. PMID- 16251752 TI - Characterization of the bovine placenta by cytoskeleton, integrin receptors, and extracellular matrix. AB - The cytoskeleton together with integrin receptors and proteins of the extracellular matrix provide sensitive indices of the development and organization of the bovine placenta. The bovine placenta is classified as synepitheliochorial because migrating trophoblast giant cells fuse with single uterine epithelial cells. This phenomenon may be interpreted as a restricted trophoblast invasion. Bovine placentomes from early placentation until term can be characterized by indirect immunohistochemical methods. In order to do so, placental tissues are snap-frozen in liquid nitrogen or perfusion-fixed in formalin and embedded in paraffin. Depending on the antibodies used, the different cell types within the cow placenta are identified either on frozen sections or on paraffin sections according to the expression of different cytoskeletal filaments (alpha smooth muscle actin, different cytokeratins, desmin and vimentin). The specific expression of integrin receptors (subunits alpha1, alpha2, alpha3, alpha4, alpha5, alpha6, alpha(v), beta1, beta3, and alpha4) as well as proteins of the extracellular matrix (collagens type I and IV, fibronectin, and laminin) in the different cell populations is also examined. PMID- 16251753 TI - Molecular markers for human placental investigation. AB - The human placenta is a source for a variety of growth factors, hormones, and other proteins. The cellular source of the proteins can be best determined by immunohistochemistry. Furthermore, immunohistochemistry can also be used to identify a specific cell type and to differentiate it from other types of cells. Thus, there is the need for specific markers of those cell types that are present in the placenta. In this chapter, the basic protocols for the identification of proteins in a tissue section are described. This chapter focuses on methods that are available in the majority of laboratories, and therefore concentrates on methods that are used together with light microscopy. PMID- 16251754 TI - Correlative microscopy of ultrathin cryosections in placental research. AB - In this chapter, we describe procedures for correlative microscopy in immunocytochemical studies on the human placenta. We have adapted ultrathin cryosections for use in high-resolution immunofluorescence microscopy (IFM) and for correlative immunocytochemical localization using fluorescence and electron microscopy. High-resolution IFM of ultrathin cryosections (50-100 nm in thickness) can be important because these physical sections minimize the potential for false co-localization in the z-dimension. In addition, IFM of these sections affords greater sampling efficiency than does immunoelectron microscopy (IEM). These ultrathin cryosections are compatible with conventional electron microscopy because a relatively low-voltage electron beam can penetrate them. Thus, the same ultrathin cryosections of placenta can be viewed in both fluorescence and electron microscopes. This latter point can be of importance because it may be necessary to know the true size and shape of objects observed by IFM; this can be determined best by IEM. Additionally, IEM can provide the "reference space" lacking in IFM. The use of ultrathin cryosections is a powerful approach for placental research, especially for the investigation of the in situ localization of antigens in the complex structure of the human placenta. PMID- 16251755 TI - Vascular corrosion casting of the uteroplacental and fetoplacental vasculature in mice. AB - This chapter describes methods for making vascular corrosion casts of the uteroplacental and fetoplacental vasculature of the mouse placenta. A catheter placed in the ascending thoracic aorta of a pregnant mouse permits the introduction of a methyl methacrylate casting compound into the lower body vasculature, including the uterus and placenta. A fine-tipped glass cannula attached to a double-lumen catheter is used to instill the same casting compound in the fetoplacental vessels of mouse placentas. Following polymerization of the casting compound, tissue is digested off of the placental casts using 20% KOH. The washed and dried casts are then available for light or scanning electron microscopy. The methods described have been used to cast the mouse uteroplacental vasculature from 5.5-18.5 d gestation and the fetoplacental vasculature from 12.5 d gestation to term. PMID- 16251756 TI - Analysis of fetal and maternal microvasculature in ruminant placentomes by corrosion casting. AB - Vascular corrosion casting is a useful tool for studying the vascular architecture of complex organs. The synepitheliochorial placenta of ruminants is composed of two closed blood circuits, a fetal and a maternal one. The microvasculature of each circuit has the shape of the corresponding cotyledon (villous trees) and caruncle (crypts). These two compartments interdigitate with each other in a complementary fashion. Understanding three-dimensional vascular arrangements is facilitated by scanning electron microscopy of vascular corrosion casts. Methods to be used in the generation of vascular casts from fetal and maternal placentomal blood vessels are described, with special emphasis on casting resins and corrosion using potassium hydroxide. The procedure of splitting larger casts following gelatin embedding and freezing is also presented. PMID- 16251757 TI - Microarray analysis of trophoblast cells. AB - A complex repertoire of trophoblast gene products governs the multifaceted functions performed by the placenta during the relatively short period of pregnancy. Cloning and sequencing the human as well as other mammalian genomes allow investigators to gain better insight into the function of trophoblast genes. Our ability to identify transcripts by their nucleotide sequences and determine their expression patterns enables us to glean information on gene function. Although the molecular principles underlying microarray are not new to biology, the high throughput, low reaction volumes, fluorescent labeling, accurate detection, and robust analysis software makes this approach most appealing to today's researchers, when compared with standard filter blotting techniques. This chapter focuses on DNA microarray of the human placental transcriptome as a means to identify alterations in gene expression in different physiological or pathological conditions. PMID- 16251758 TI - Gene expression microarray data analysis of decidual and placental cell differentiation. AB - Gene expression analysis using DNA microarray approaches have provided new insights into the physiology and pathophysiology of many biological processes. These include identification of genetic programs and pathways that underlie cell and tissue differentiation and gene expression programs responsive to genetic perturbations, drugs, toxins, and infectious agents. In this chapter, we present methods for the analysis of microarray data using earlier investigations from our laboratory as examples of how gene expression patterns for cellular differentiation may be detected and analyzed for biological significance and how regulated genes may be classified into functional categories and pathways. PMID- 16251759 TI - Assays to determine allelic usage of gene expression in the placenta. AB - Mammalian placentas express a large number of so-called imprinted genes. Imprinting refers to mono-allelic or biased expression based on which parent contributed the allele. Many of these imprinted loci encode factors involved in growth and cell-cycle regulation, as well as maternal behavior. In general, paternally expressed genes tend to enhance growth, whereas maternally expressed genes inhibit growth. Methods are described for developing assays to test the allelic usage of a gene. The approaches described are best utilized within a system where multiple strains are available, and it is possible to perform reciprocal crosses. Only polymerase chain reaction-based methods are examined in any detail. PMID- 16251760 TI - Adenoviral-mediated gene delivery to trophoblast cells. AB - This chapter focuses on technology for construction of recombinant adenoviruses containing reporter genes under the control of putative regulatory regions of the human (h)CYP19 (aromatase) gene, as well as expression vectors. These recombinant adenoviruses have been used in primary cultures of human placental cells to characterize regulatory regions of the hCYP19 gene and to analyze the function of transcription factors on hCYP19 expression and on trophoblast differentiation. PMID- 16251762 TI - Asthma adherence and the athlete. AB - Although much has been written on the topic of adherence (also known as compliance) in asthma, little is known about adherence in the asthmatic athlete. There are shared reasons for poor adherence to treatment among athletes of varying ages and levels of competition. The Shared Decision-Making model is considered to be the optimal approach to enhancing adherence among all patients with chronic disease but is particularly well applied to athletes with asthma. This article discusses athlete-related, physician-related, and treatment-related barriers to adherence, as well as how to overcome these barriers to promote optimal disease management. Special considerations for the elite or professional athlete are also addressed. PMID- 16251763 TI - Differential diagnosis of chest symptoms in the athlete. AB - Chest pain is a common complaint of athletes in all age groups. In athletes, chest pain is often attributed to "chest tightness," and treatment for bronchospasm is considered. However, the causes of the pain are wide and varied, and the pain is referable to the many organ systems that localize to the thorax. Therefore, when treatment with bronchodilators fails, it becomes important to consider other nonasthmatic causes of the pain. These causes can be organized by system and are explained in this article. Cardiac causes are the most feared and, fortunately, are very rare in the adolescent setting. With a thorough knowledge of etiologies of chest pain, the physician can often make a diagnosis with only a history and a physical exam. PMID- 16251764 TI - Medical screening of the athlete: How does asthma fit in? AB - There are 4 million high school athletes in the United States, and there are 30 million athletes under age 18 yr competing in organized sports. The prevalence number of emergency room visits and deaths from asthma are significant. Medical clearance is required by all states for participation in high school sports. This article discusses the issues regarding the medical screening of the athlete and how asthma fits into the process. The preparticipation sports physical is an overlooked and underutilized opportunity to improve asthma diagnosis and management in adolescents. PMID- 16251765 TI - Making the diagnosis of asthma in the athlete. AB - This article encompasses a discussion of the clinical presentation and features as various diagnostic modalites of bronchoprovocation and differential diagnosis used in screening for asthma in the athlete. A comparative analysis of these modalities, including questionnaire, treadmill, eucapneic hyperventilation, as well as field challenge, is provided with sensitivities and specificities, predictive value, and application to clinical practice. The indication and mechanics of each diagnostic modality is discussed, providing the clinician with a comprehensive understanding of the principles and procedures used to diagnosis asthma in the athlete. PMID- 16251766 TI - Asthma deaths and the athlete. AB - Asthma has recently become more prevalent, more severe, and more deadly. Approximately 4500 individuals die from asthma annually in the United States, an unacceptable number for a condition that can be managed effectively. Sudden death with exercise may result from a variety of causes, including previously unrecognized cardiac conditions. Asthma has also been recognized as a cause of death in association with sports. Recent data indicate those who suffer from mild to moderate asthma are also at risk for asthma fatality. The absolute magnitude of the increase in risk of death from asthma during sports, however, is very small. For this reason, individuals with asthma should not be discouraged from active participation in sports. Rather, this should reinforce the message that asthma is a condition that may be potentially serious, but can, and should be, well controlled with proper management. PMID- 16251767 TI - Asthma and the diver. AB - Self-contained underwater breathing apparatus (scuba) diving has grown in popularity, with nearly 9 million sport divers in the United States alone. Approximately 7% of the population has been diagnosed with asthma, which is similar to the percentage of divers admitting they have asthma. Numerous concerns exist regarding subjects with asthma who choose to participate in recreational diving. Among these concerns are pulmonary barotrauma, pneumomediastinum, pneumothorax, arterial gas embolism, ear barotrauma, sinus barotrauma, and dental barotrauma. Despite these concerns, a paucity of information exists linking asthma to increased risk of diving complications. However, it has long been the norm to discourage individuals with asthma from participating in recreational scuba diving. This article examines the currently available literature to allow for a more informed decision regarding the possible risks associated with diving and asthma. It examines the underlying physiological principles associated with diving, including Henry's law and Boyle's law, to provide a more intimate understanding on physiological changes occurring in the respiratory system under compressive stress. Finally, this article offers a framework for guiding the patient with asthma who is interested in scuba diving. Under the right circumstances, the patient with asthma can safely participate in recreational diving without apparent increased risk of an asthma-related event. PMID- 16251768 TI - Asthma and athletes: therapy to compete. AB - Asthma presents special challenges to both the athletes who have it and to their health care providers. This article briefly reviews the problem of asthma- especially exercise-induced asthma--in the competitive athlete, and then describes treatments that are effective in controlling asthma. Drug-doping regulations are explained, as is the worldwide impact of drug doping on competitive athletes who have asthma. This review concludes with recommendations for competitive athletes and their health care providers regarding how to deal with asthma in this patient population. PMID- 16251769 TI - Nasal obstruction, the airway, and the athlete. AB - Rhinitis is a common condition that affects a significant proportion of the general population, as well as a high proportion of athletes. Nasal congestion is a predominate symptom of the late-phase reaction in allergic rhinitis and can have far-reaching effects that extend through the airway and beyond the nose. Rhinitis is often found in conjunction with asthma and is a risk factor for asthma. Nasal obstruction, which does not permit conditioning of inspired air by the nasal turbinates, may contribute to asthma symptoms and the development of asthma. These adverse conditions may be especially troublesome for the high performance athlete who has increased nasal airflow turbulence and who competes under extreme conditions that may worsen rhinitis and asthma. Under the theory of the unified airway, an immune response induced in the nose may extend into the lungs via cytokines and other inflammatory mediators. Nasal congestion can significantly contribute to sleep dysfunction, leading to daytime fatigue and decreased performance. Treatment of allergic rhinitis can improve sleep and foster productivity. Control of rhinitis and nasal congestion, which is obtained by various therapies, may reverse lower airway tendency to bronchoconstriction. PMID- 16251771 TI - Healthy weight, healthy shape. PMID- 16251770 TI - Asthma and activities of daily living. AB - The provider's ability to judge the severity of a patient's illness is a vital part of the care of chronic disorders. In the treatment of asthma, it should extend beyond the objectively quantifiable measures of obstruction or inflammation. "Interference with activities of daily living" is a highly subjective assessment that may not allow quantitative comparisons, but it matters to the patient. Interventions aimed at lessening the burden of asthma on the daily life of the patient and his or her family may hold the key to improvement in a broader range of outcome measures. PMID- 16251772 TI - Coronary risk variables in young asymptomatic smokers. PMID- 16251773 TI - Hantaviruses are here in India. PMID- 16251774 TI - Optimizing treatment in head and neck cancers -- are molecular markers the answer? PMID- 16251775 TI - Endosymbiotic Wolbachia of parasitic filarial nematodes as drug targets. AB - The parasitic nematodes Wuchereria bancrofti, Brugia malayi and B. timori cause a dreadful disease in humans known as lymphatic filariasis, which afflicts more than 120 million people worldwide. As per recent epidemiologic estimates on prevalence of W. bancrofti and B. malayi, about 428 million people are at risk, with 28 million microfilaria carriers and 21 million clinical cases spread out in 13 States and 5 Union Territories of India. The Indian subcontinent that comprises Bangladesh, India, Maldives, Nepal and Sri Lanka harbours 50 per cent of the world's lymphatic filarial disease burden. Recently, an endobacterium of Wolbachia species that belongs to the family Rickettsiaceae was found in all life cycle stages of these nematodes and the transmission is exclusively vertical through the embryonic stages of the female worms. People with filariasis have been exposed to these Wolbachia bacteria or their proteins by the natural killing of parasites. Wolbachia have also been identified occasionally in body fluids of infected patients. Evidence suggests that these Wolbachia are mutualistic symbionts and can be cured from the nematodes by several antibiotics having antirickettsial properties. Treatment of nematodes with tetracyclines affect Wolbachia and they get cleared from worm tissues; and this elimination causes reproductive abnormalities in worms and affect worm's embryogenesis, resulting in sterility. Although it is impractical, prolonged treatment with doxycycline significantly reduces the numbers of microfilaria in circulation, which is an important strategy to control transmission of filariasis by mosquito vectors. In this review, the current knowledge of Wolbachia as a drug target and potential ways to reduce the infection through anti-Wolbachia treatments is discussed. PMID- 16251776 TI - Coronary risk variables in young asymptomatic smokers. AB - BACKGROUND AND OBJECTIVES: Smoking plays a dominant role in premature atherosclerosis particularly among males in South Asian countries. It initiates and promotes atherosclerosis by altering cardiac haemodynamics, causing dyslipidaemia and producing oxidative damage. Not much information is available from our country. We therefore undertook this study to see the effect of smoking on electrocardiogram (ECG), blood pressure, lipids, apolipoprotein B level and free radical activity in young asymptomatic male smokers. METHODS: The study included 100 consecutive male subjects (50 smokers and 50 non smokers) aged 30-40 yr. Smoking profile, detailed cardiovascular assessment including ECG and lipid profile were evaluated in each subject. RESULTS: Of the 50 smokers, 22 (44%) had grade I hypertension as against 5 of 50 non smokers. Sinus tachycardia (10%) and P-pulmonale (8%) were the only notable ECG abnormalities. Dyslipidaemia was detected in 92 per cent smokers and 48 per cent non smokers (P<0.001). Total serum cholesterol, low density lipoprotein (LDL)-cholesterol, triglycerides and apolipoprotein B levels were significantly higher (P<0.001) in smokers compared to non smokers. LDL-cholesterol was > or =135 mg/dl in 94 per cent dyslipidaemic smokers. However, no significant difference was found in high density lipoprotein (HDL)-cholesterol. Smokers had significantly higher serum malondialdehyde levels (P<0.001) and low superoxide dismutase (P<0.001) compared to non smokers. INTERPRETATION AND CONCLUSION: Our data indicate that young asymptomatic male smokers tend to have hypertension, dyslipidaemia and increased production of free oxygen radicals, perhaps by attenuation of oxidative stress by cigarette smoking. This makes them prone for premature coronary artery disease. However, the findings need to be confirmed on a larger sample. PMID- 16251777 TI - A pilot study for serological evidence of hantavirus infection in human population in south India. AB - BACKGROUND AND OBJECTIVES: Hantaviruses are rodent-borne viruses of the family Bunyaviridae that have been identified as aetiological agents of two human diseases, haemorrhagic fever with renal syndrome (HFRS) and hantavirus pulmonary syndrome (HPS). There are no reports of hantavirus infections in humans from India, hence this pilot study was undertaken to provide the serological evidence of hantavirus infections in humans in south India. METHODS: Serum samples were obtained from individuals with acute febrile illness and from voluntary blood donors, majority of whom were from south India. Serum samples were tested for anti-hantavirus IgM using a commercial enzyme immunoassay (EIA). Samples found positive by the EIA were tested by an indirect immunofluorescence assay (IFA) using slides coated with Seoul virus (SEOV) infected cells as substrate. RESULTS: Of the 152 serum samples from individuals with pyrexic illness, 23 (14.7%) were positive for anti-hantavirus IgM by EIA. In contrast, only 5.7 per cent of healthy blood donors were positive by this assay. Eighteen of the 22 (82%) EIA positive samples from patients were positive by the IFA assay. In contrast, only 2 of the 5 (40%) blood donor EIA positive samples were positive in the IFA assay. INTERPRETATION AND CONCLUSION: The finding of this study indicated the possible presence of hantavirus infections in the human population of India presenting both as asymptomatic and symptomatic infections. Further studies need to be done to confirm the findings on a larger sample using molecular techniques. PMID- 16251778 TI - Association of DNA pattern of metastatic lymph node with disease free survival in patients with intraoral squamous cell carcinoma. AB - BACKGROUND AND OBJECTIVE: Intraoral squamous cell carcinoma (OSCC) is one of the common tobacco related cancers affecting Indian population. These tumours are slow growing, endophytic and are mostly well differentiated. Cervical lymph node is the common site of metastasis of these tumours. In most of the patients cervical lymph node metastasis rather than the primary tumour, affects prognosis. However, no reports are available on the DNA pattern of the metastatic lymph nodes in patients with intraoral squamous cell carcinoma. Therefore, the present study was undertaken to observe DNA pattern of primary tumours and their corresponding metastatic lymph nodes and its association with the clinicopathological parameters and prognosis. METHODS: DNA flow cytometry was successfully carried out on 68 paraffin embedded specimens of the primary tumours and their 22 corresponding metastatic cervical lymph nodes. The findings were evaluated for their possible association with clinicopathological features of the tumour and disease free survival of patients with intraoral carcinoma. RESULTS: Analysis of nuclear DNA patterns revealed 32 (47.0%) diploidy and 36 (52.9%) aneuploidy in primary tumours whereas metastatic lymph nodes showed 7 (31.8%) diploidy and 15 (68.1%) aneuploidy. The aneuploidy group in metastatic lymph node had significantly higher S phase fraction (SPF) (P<0.01) and poor histological grade (P<0.002) as compared to their counterparts with diploidy. DNA pattern of metastatic lymph node further showed a significant association with disease free survival in the log rank test. Aneuploidy and high SPF in metastatic lymph node was found to be associated with early recurrence while DNA pattern of the primary tumour did not show significant association with the disease free survival. INTERPRETATION AND CONCLUSION: It may be concluded that aneuploidy and high SPF in metastatic lymph node might be considered as an important discriminatory risk factor in patients with similarly staged intraoral squamous cell carcinoma. PMID- 16251779 TI - AmpC beta-lactamase producing bacterial isolates from Kolkata hospital. AB - BACKGROUND AND OBJECTIVE: The widespread use of beta-lactam antibiotics has lead to the development of resistance to this group of antibiotics in bacterial pathogens due to beta-lactamase production. Information on such pathogens is not available from eastern region of India. This study was undertaken to determine the AmpC beta-lactamase production in pathogens isolated from hospitalized patients in Kolkata. METHODS: Non-repeat clinical isolates (284) from pus, urine, sputum and other clinical specimens of hospitalized patients were taken. Disk agar diffusion (DAD) and minimum inhibitory concentration (MIC) with different beta-lactam antibiotics, and double disc synergy test (DDST) with clavulanic acid and sulbactam were done. Disk antagonism test (DAT) and three-dimensional extract test (TDET) were conducted for phenotypic confirmation of AmpC and inducible AmpC beta-lactamase production. Nitrocefin spot test and microiodometric assay of beta lactamase were also performed. RESULTS: Twenty seven isolates were found to be resistant to cefoxitin, a alpha-methoxy-beta-lactam. Of these, 19 were observed to be AmpC beta-lactamase producers and 4 were inducible AmpC beta- lactamase producers by DDST, DAT and TDET. Remaining 4 were non AmpC beta-lactamase producers. Of the 23 AmpC beta-lactamase producers, the distribution of different species was as follows: Escherichia coli 11 (47.8%), Pseudomonas aeruginosa 4 (17.3%) Klebsiella pneumoniae 3 (13%), and Klebsiella aeruginosa 1 (4.3%). INTERPRETATION AND CONCLUSION: Our finding showed 6.7 per cent AmpC beta lactamase and 1.4 per cent inducible AmpC beta-lactamase producing clinical isolates from Kolkata. AmpC beta-lactamase producing bacterial pathogens may cause a major therapeutic failure if not detected and reported in time. PMID- 16251780 TI - Prevalence of HIV and VDRL seropositivity in blood donors of Delhi. AB - Blood transfusion has been the transmission mechanism in 15 per cent of total patients infected with human immunodeficiency virus (HIV). A few reports are available regarding the trend of HIV seropositivity in northern India. Prevalence of VDRL (venereal disease research laboratory) reactivity varies from 0.8-15 per cent in blood donors. We present data on the prevalence and trends of infection with HIV 1 and 2 and VDRL reactivity in blood donors in Delhi. Between 2000-2002, a total of 76089 (voluntary and replacement) donors were screened. Majority (82.4%) were replacement donors. Seropositivity for HIV and VDRL was seen in 0.54 and 2.6 per cent of donors respectively. The percentage of seropositivity for HIV and VDRL was significantly higher in replacement donors (P<0.001). It is suggested that extensive donor selection and a voluntary donor service would reduce the number of infectious donors significantly. Non-renumerated repeat voluntary donor services are urgently required to lower the prevalence of transmissible infections. While the need to change to a voluntary donor service and devising effective donor screening cannot be over emphasized, there is also a need to mandate HIV antigen detection in India. PMID- 16251781 TI - Isolation, phage typing and antibiogram of Salmonella from man and animals in northeastern India. AB - BACKGROUND AND OBJECTIVE: Salmonella is an important zoonotic pathogen and its prevalence in the animals acts as a continuous threat to man. The present study was carried out to report the isolation along with the serotypes, phage types and antibiogram pattern of Salmonella among man, livestock and poultry in the northeastern India. METHODS: A total of 654 samples from diarrhoeic livestock and humans were processed for the isolation of Salmonella. All the isolates were subjected to antibiogram studies against 15 antimicrobials. Representative isolates of S. Typhimurium and S. Enteritidis were phage typed. RESULTS: Ninety five isolates of Salmonella enterica belonging to 5 serotypes- S. Typhimurium, S. Enteritidis, S. Gallinarum, S. Paratyphi B and S. Bareilly were obtained with an overall prevalence rate of 14.40 per cent. S. Typhimurium isolates were distributed among four phages- DT003, DT004, DT096 and DT193 and all the S. Enteritidis isolates belonged to a single phage type, PT13a/7. Interspecies sharing of the phages was observed. Norfloxacin, enrofloxacin, gentamycin and ciprofloxacin were most effective, whereas, doxycycline, ampicillin, amoxycillin and tetracycline were relatively less effective. INTERPRETATION AND CONCLUSION: Our findings showed that three of the five serovars as well as some of the phage types of these serovars were shared by animals and humans indicating the zoonotic potential of the organism. Thus, it is imperative that salmonellosis control measures adopted for humans should give adequate importance to its control in the animals particularly their products. PMID- 16251782 TI - Estimation of burden of tuberculosis in India for the year 2000. AB - BACKGROUND AND OBJECTIVE: Data on the burden of tuberculosis (TB) in India are vital for programme planners to plan the resource requirements and for monitoring the nation-wide TB control programme. There was a need to revise the earlier estimate on the burden of TB in India based on the increase in population and current epidemiological data. This study estimates the burden of disease for the year 2000 based on recent prevalence of TB and annual risk of tuberculosis infection (ARTI) estimates. METHODS: Data on prevalence generated among adults by the Tuberculosis Research Centre (TRC), Chennai, among children by National Tuberculosis Institute (NTI), Bangalore, and the ARTI estimates from the nation wide sample survey by NTI and TRC were used for the estimation. The prevalence of disease corresponding to 1 per cent ARTI was extrapolated to different parts of the country using the estimates of ARTI and the population in those areas and added together to get the total cases. Abacillary cases that required treatment were estimated from X-ray abnormals. The estimates of bacillary, abacillary and extrapulmonary cases were then combined to get the national burden. RESULTS: The estimated number of bacillary cases was 3.8 million (95% CI: 2.8-4.7). The number of abacillary cases was estimated to be 3.9 million and that for extrapulmonary cases was 0.8 million giving a total burden of 8.5 million (95% CI: 6.3-10.4) for 2000. INTERPRETATION AND CONCLUSION: The present estimate differs from the earlier estimates because we have included the disease burden of X-ray cases that are likely to breakdown to bacillary cases in a one year period, and extrapulmonary TB cases. The current estimates provided baseline information for advocacy and planning resource allocation for TB control activities. Also, these estimates can be compared with that in future years to measure the long term impact of TB control activities in India. PMID- 16251783 TI - Neurosyphilis in a tertiary care hospital in north India. AB - BACKGROUND AND OBJECTIVES: The clinical diagnosis of neurosyphilis is very rarely encountered today in the developed world although syphilis remains a significant health problem in few areas of the industrialized countries and in most of the third world nations. This apparent decline may be due to increase in number of asymptomatic neurosyphilis and cases presenting as subtle, illdefined syndromes rather than classic presentation of tabes dorsalis and general paresis in the post penicillin era. This retrospective study was carried out to report the neurosyphilis cases diagnosed at a tertiary care hospital in North India, and to analyse the laboratory and clinical parameters of these cases. METHODS: Suspected cases of neurosyphilis presenting at Postgraduate Institute of Medical Education and Research (PGIMER), Chandigarh over a period of 13 yr (January 1990 to December 2002) were identified. Diagnosis of neurosyphilis was based on clinical presentation, prior history of syphilis, routine CSF biochemistry (protein and leukocytes) and serological evidence [serum and CSF venereal disease research laboratory (VDRL) and Treponema pallidum particle agglutination (TPPA) tests]. RESULTS: A total of 25 cases of neurosyphilis were identified, 18 (72%) with reactive CSF-VDRL, 22 (88%) with elevated CSF protein and 24 (96%) with CSF mononuclear leukocytosis. Serum VDRL was reactive in all 25 cases. Three patients were asymptomatic (2 primary syphilis; 1 early latent stage), 8 had secondary and 14 had tertiary syphilis. Two of the neurosyphilis cases were also seropositive for HIV. Radiology was abnormal in 7 (28%) patients. INTERPRETATION AND CONCLUSION: Neurosyphilis still remains a problem in a country like India and a high index of suspicion and clinical expertise are required for appropriate diagnosis and proper management especially in the era of AIDS pandemic. PMID- 16251784 TI - Seroprevalence of hepatitis D virus in patients with hepatitis B virus-related liver diseases. AB - BACKGROUND AND OBJECTIVE: Several reports indicated a declining trend in the occurrence of hepatitis D virus (HDV) infection in some geographical areas. However, no study has been conducted in India to evaluate whether a similar epidemiological change is occurring in this part of the world. The present study was undertaken to evaluate the seroprevalence of HDV in patients with hepatitis B virus (HBV) related liver diseases attending a Government hospital in New Delhi, and to assess any change in its epidemiology by comparing the results with seroprevalence figures reported in the past. METHODS: A total of 123 patients with HBV-related liver diseases comprising 32 cases of acute viral hepatitis (AVH), 5 of fulminant hepatic failure (FHF), 37 of chronic hepatitis (CH), 46 of cirrhosis and 3 of hepatocellular carcinoma (HCC). All patients were evaluated for the presence of delta antibodies using commercially available ELISA kits. Both IgM and IgG anti-delta assays were performed to differentiate between active and convalescent infection. RESULTS: The mean age of the patients was 35.6 +/- 3.3 yr with a male : female ratio of 11:5. Of the 123 patients, serological evidence of delta virus infection was seen in 13 subjects (10.6%); 9 (7.3%) had evidence of past infection (IgG positive, IgM negative) and the remaining 4 (3.3%) recent infection (IgM anti-delta antibody positive). Evidence of HDV infection in acute viral hepatitis, fulminant hepatitis, chronic hepatitis, cirrhosis and hepatocellular carcinoma groups was found in 3.1, 20, 8.1, 15.2 and 33.3 patients, respectively. INTERPRETATION AND CONCLUSION: Our results suggest that delta infection may not be very common in Indian patients with HBV-related liver diseases. It is also possible that HDV epidemiology in this part of the world may be undergoing a transition towards decreasing prevalence. PMID- 16251785 TI - HIV-related knowledge, attitudes and risk perception amongst nurses, doctors and other healthcare workers in rural India. AB - BACKGROUND AND OBJECTIVE: People with HIV in India frequently encounter discrimination while seeking and receiving healthcare services. The knowledge and attitudes of healthcare workers (HCWs) influences the willingness and ability of people with HIV to access care, and the quality of the care they receive. Previous studies of HIV-related knowledge and attitudes amongst Indian HCWs have been conducted primarily in large urban hospitals. The objective of this study was to asses HIV-related knowledge, attitudes and risk perception among a group of rural north Indian HCWs, and to identify predictors of willingness to provide care for patients with HIV infection. METHODS: A cross-sectional survey of 266 HCWs (78% female) from seven rural north Indian health settings was undertaken in late 2002. A self-administered written questionnaire was made available in English and Hindi, and the response rate was 87 per cent. Information was gathered regarding demographic details (age, sex, duration of employment, job category); HIV-related knowledge and attitudes; risk perception; and previous experience caring for HIV-positive patients. Logistic regression modelling was undertaken to identify factors associated with willingness to care for patients with HIV. RESULTS: The HCWs in this study generally had a positive attitude to caring for people with HIV. However, this was tempered by substantial concerns about providing care, and the risk of occupational infection with HIV was perceived by most HCWs to be high. After controlling for confounding, HCWs willingness to provide care for patients with HIV was strongly associated with having previously cared for patients with HIV (P = 0.001). Knowledge of HIV transmission and perception of risk were not associated with willingness to provide care. INTERPRETATION AND CONCLUSION: The findings of this study showed a general willingness of HCWs to provide care for patients with HIV, tempered by concerns regarding provision of such care. Strategies to address HCWs concerns are likely to ameliorate the discrimination experienced by people with HIV when accessing healthcare services. These include the development of programmes to promote occupational safety of HCWs and involving people with HIV in awareness training of HCWs. PMID- 16251786 TI - Severe vitamin A deficiency in India during pulse polio immunization. PMID- 16251787 TI - Malnutrition and dental caries: a review of the literature. AB - Protein-energy malnutrition occurs when there are deficiencies in protein, energy foods or both, relative to a body's needs. This paper reviews the association of early childhood malnutrition with: (1) dental caries, (2) enamel hypoplasia, (3) salivary gland hypofunction, and (4) delayed eruption. Studies suggest that caries of the primary dentition is associated with early childhood malnutrition, though the effect on caries of the permanent dentition has essentially not been studied. Enamel hypoplasia, salivary glandular hypofunction and saliva compositional changes may be mechanisms through which malnutrition is associated with caries, while altered eruption timing may create a challenge in the analysis of age-specific caries rates. PMID- 16251788 TI - A mouse caries model and evaluation of aqp5-/- knockout mice. AB - Current techniques to alter gene expression in mice allow direct analysis of the net role of a host factor in caries development. Towards this goal we first established protocols to induce and score caries in NFS/N mice and determined caries susceptibility in mice with targeted deletion of the gene encoding aquaporin-5 (Aqp5-/-), a water channel involved in the production of saliva. In the NFS/N strain of mice total sulcal caries and severity scores were consistent between experiments, whereas smooth surface caries scores were lower, more variable but distributed fairly evenly among the buccal, lingual and sulcal surfaces. In Black Swiss/129SvJ mice (genetic background of Aqp5-/- mice) caries scores were 50-75% lower compared to NFS/N mice, suggesting strain variation in caries susceptibility under our experimental conditions. In Aqp5-/- mice, in which the volume of total salivary secretion is reduced by 60-65%, there was a significant increase in caries, primarily on the buccal and sulcal surfaces. Results indicate that caries susceptibility increases with a reduced salivary flow that is associated with decreased water content of saliva. PMID- 16251789 TI - Outcomes of a non-operative caries treatment programme for children and adolescents. AB - This study assessed the effectiveness and performance of a non-operative caries treatment programme (NOCTP) used since 1987 in the municipality of Nexo in Denmark. The NOCTP emphasizes mechanical plaque control and considers the eruption period of molar teeth as a risk factor. The mean DMF-S among 18-year olds in 1999 and 2000 in Nexo was 1.23 +/- 2.26 and 1.25 +/- 2.01 (medians 0); 55 and 56% had DMF-S = 0. The mean numbers of sealed surfaces were 4.6 +/- 3.25 and 4.0 +/- 3.22, respectively. The cost per child per year was marginally and significantly reduced in the years with the NOCTP compared to that before 1988 (p = 0.05). In 4 comparison municipalities with very low caries experience, mean DMF S scores among 18-year-olds in 1999 were 2.73-3.25 (medians 1-2) and were significantly higher than in Nexo (p < 0.001). The NOCTP differed from the preventive programmes used in the comparison municipalities in the period 1988 1999 in emphasising care for the erupting molars, the use of a firm guideline and stated goals to be achieved, but with less emphasis on diet. The effectiveness and performance of the NOCTP were both considered high, as very low DMF-S and high %DMF-S = 0 had been achieved by 1999, and 18-year-olds in Nexo had significantly less caries than in the comparison municipalities. The latter difference could not be explained by difference in caries-related background variables. PMID- 16251791 TI - The association of acidic reflux above the upper oesophageal sphincter with palatal tooth wear. AB - This case-controlled clinical study, conducted at a secondary care unit, compared pH measurement at 2 cm above the upper oesophageal sphincter (UOS) in 31 patients complaining of extra-oesophageal symptoms of reflux to 7 control subjects. Twenty four-hour ambulatory pH measurements were recorded and analysed at 5 cm above the lower oesophageal sphincter (LOS) and 2 cm above the UOS. In reflux patients the proportion of supine time when pH at 5 cm above the LOS was <4 was significantly greater than in controls. The proportion of time when pH at 2 cm above the UOS was <5.5 was also significantly greater in patients than in controls. Palatal tooth wear observed in the patient group correlated with acid reflux at night. In this group of patients presenting with symptoms of reflux, gastric acid passed through the upper oesophageal sphincter and increased the potential for erosion. PMID- 16251790 TI - Effects of sucking acidic candy on whole-mouth saliva composition. AB - Limited information is available on the effects of sucking acidic candies on saliva composition and the protective role of saliva in this relation. Therefore the aim of this study was to determine salivary effects of sucking acidic candies in vivo in relation to individual variations in whole-saliva flow rate (WSFR) and buffer capacity (WSbeta). Ten healthy young males (24 +/- 2 years) sucked a rhubarb-flavoured acidic hard-boiled candy with tartaric acid available on the Danish market. The whole saliva was collected into a closed system, regarding CO2, at different times as follows: firstly, unstimulated saliva for 5 min (baseline), secondly stimulated saliva for 4 min upon sucking the candy, and finally post-stimulated saliva for 10 min. Saliva pH was determined on a blood gas analyser and WSbeta was estimated from the saliva bicarbonate concentration obtained by the analyser and by ionic balance calculation. The erosive potential of the candy in saliva was estimated from the saliva pH values and degree of saturation with respect to hydroxyapatite (DS(HAp)). The results showed that saliva pH dropped from 6.5 (baseline) down to 4.5 at the fourth minute of sucking the candy, and returned to pH 6.5 five minutes after stimulation (post stimulated). DS(HAp) decreased upon sucking the candy and saliva from all subjects became undersaturated with respect to HAp. Significant positive correlations were obtained between pH and WSFR (r(s) = 0.47; p < 0.05) and between pH and WSbeta (r(s) = 0.65; p < 0.01). In relation to WSbeta we found that 70% of the buffer capacity originating from the bicarbonate buffer system upon sucking the candy was exerted as phase buffering. We conclude that sucking this type of acidic candies changes whole-mouth saliva composition so that it may have erosive potential and that high WSFR and WSbeta have protective effects against these salivary changes. PMID- 16251792 TI - Cariogenicity of the probiotic bacterium Lactobacillus salivarius in rats. AB - Probiotic bacteria such as lactobacilli and bifidobacteria are considered to be non-pathogenic and non-toxigenic on the basis of long years of safe usage. However, some species of lactobacilli are thought to be associated with the development of dental caries. The purpose of the present study was to examine the cariogenicity of the probiotic bacterium Lactobacillus salivarius in rats. Rats were divided into six groups, and infected with L. salivarius LS1952R and/or Streptococcus mutans MT8148R. L. salivarius LS1952R became established in the oral cavity of rats and induced significant level of dental caries even when infected for only 5 days from 18 to 22 days of age. In addition, the caries scores of rats superinfected with both Streptococcus mutans MT8148R and L. salivarius LS1952R from 18 days of age were significantly higher than those infected with either L. salivarius LS1952R or S. mutans MT8148R alone. Since strain LS1952R can adhere to saliva-coated hydroxyapatites, it is concluded that L. salivarius strain LS1952R possesses an inherent cariogenic activity following adherence to the tooth surface. PMID- 16251793 TI - The persistence of Streptococcus mutans in nasopharyngeal carcinoma patients after radiotherapy. AB - The aim of the study was to examine the persistence of oral Streptococcus mutans in nasopharyngeal carcinoma patients after radiotherapy. Ten subjects, ranging in age from 20 to 67 years, participated. DMFT/DMFS, salivary level of mutans streptococci and oral health status were recorded. Pooled plaque samples were obtained from the cervical margins and the interproximal regions of all the teeth and the occlusal surfaces of the molars prior to, immediately after, 3 and 6 months after the completion of radiotherapy. At least 10 colonies of S. mutans were isolated from each subject and totally 645 isolates were genotyped by restriction endonuclease analysis. The results showed that the salivary level of S. mutans increased significantly with the reduction of salivary flow rate after radiotherapy. Each subject had at least 1 genotype of S. mutans isolated throughout the follow-up period. In 3 subjects who initially carried 2 or more genotypes, 1 or 2 genotypes of S. mutans could not be detected 3 months after treatment. Moreover, the genotypes that became undetectable were predominant bacteria in the first sampling. The result indicated that most S. mutans genotypes were persistent after radiotherapy but some genotypes that might not adapt to the alteration of oral environment became undetectable. PMID- 16251794 TI - Clinical effectiveness of laser fluorescence, visual inspection and radiography in the detection of occlusal caries. AB - The aim of this in vivo study was to compare a laser fluorescence (LF) device with Ekstrand's visual scoring system and radiographic assessment for detection of occlusal caries. Thirty-eight adults aged 19-35 years participated in the study; a total of 57 third molars with macroscopically intact occlusal surfaces were selected. Two examiners assessed 110 sites by visual inspection (VI), bitewing radiography (BW) and LF. Teeth were then extracted and caries extent assessed by histology. The detection methods were compared by means of sensitivity, specificity, inter-examiner reproducibility (kappa statistics) and area under the ROC curve. VI and LF had similar (p > 0.05) and superior sensitivities than BW (p < 0.05). VI and BW showed similar specificities, which was superior to LF. The inter-examiner reproducibility was good for VI and BW and moderate for LF. The area under ROC curves showed that VI was better than LF. It was concluded that Ekstrand's visual scoring system is the most valid method for caries diagnosis. LF should be considered an adjunct to caries diagnosis. PMID- 16251795 TI - Evaluation of selective caries removal by a fluorescence feedback-controlled Er:YAG laser in vitro. AB - AIM: To establish a fluorescence threshold level that could guide a therapeutic Er:YAG laser through a caries lesion to determine a therapeutic endpoint of caries removal. MATERIALS AND METHODS: A total of 65 extracted human teeth, 35 with dentine caries and 30 healthy, were used for this study. An Er:YAG laser system that emitted at a wavelength of 2.94 microm was used. The laser was equipped with a laser fluorescence feedback system, excitation wavelength 655 nm, to control the irradiation by the Er:YAG laser. The evaluated threshold levels of the fluorescence feedback system were 3, 7, 8, 10, 12, 15 and 20. After treatment the teeth were prepared for histological staining according to the method of Brown and Brenn for the identification of bacteria. The specimens were subjected to a quantitative evaluation of residual bacteria on the treated dentine surface. In addition, the internal fluorescence of dentine and potential fluorescence changes of dentine after laser irradiation were evaluated. RESULTS: About 80% of the irradiated dentine surface showed residual bacteria with threshold levels of 20, 15, 12, and 10. Residual bacteria were not found with threshold levels of 7 and 3. The study revealed a significant increase in dentine fluorescence after laser irradiation. CONCLUSION: The results of the present in vitro study indicate that a fluorescence threshold level of 7 or 8 units can guide an Er:YAG laser to a complete removal of carious dentine. PMID- 16251796 TI - Fluoride concentrations in enamel and dentin of primary teeth after pre- and postnatal fluoride exposure. AB - The aim of this study was to determine any existing difference in the amount of fluoride incorporated in the surface, body enamel and dentin of two groups of deciduous teeth, either exposed to pre- and postnatal fluoride supplements or to postnatal fluoride only. One hundred and eighty five subjects with intact deciduous incisors were selected from a randomized, double blind study of the caries preventive efficacy of prenatal fluoride (F) supplementation. Surface and body enamel samples were obtained by the acid etch biopsy technique. Dentin microsamples were obtained by drilling to a depth of 100 microm using the microdrill biopsy technique. It was concluded that fluoride exposure during the prenatal period offered no additional measurable fluoride uptake by dental tissues other than that attributable to postnatal fluoride alone. PMID- 16251797 TI - Effect of fluoride-releasing filling materials on underlying dentinal lesions in vitro. AB - Fluoride-releasing materials placed over carious tissue are assumed to enhance remineralisation of the underlying lesion. This remineralisation, however, also depends on the availability of calcium and phosphate, which may be supplied by the pulpal fluid. The aim of this study was to measure the fluoride release of glass ionomer cements (GICs) into underlying dentin and to measure the effect of the released fluoride on the remineralisation of the underlying dentinal lesions using transversal microradiography. Discs of fluoride-releasing GIC were placed on top of dentinal lesions in an in vitro model. The discs and the dentin slabs were covered completely by a protective layer of nail varnish, leaving only the pulpal side of the dentin slab open, and hence the dentinal tubules as the pathway for the incubation fluid to the GIC disc. Specimens were incubated in a remineralisation buffer. The materials tested were a conventional GIC, an experimental GIC that was designed to have a high fluoride release, and an inert material. Fluoride was found to penetrate through the dentin slab into the surrounding fluid. Fluoride uptake from the experimental GIC was higher than from the conventional GIC. Mineral content-depth profiles after 10 weeks' remineralisation revealed that in the outer 30 microm of the lesion a higher mineral deposition occurred for the experimental GIC than in both other groups. No differences in the overall change of integrated mineral loss were found for the tested materials. We conclude that high fluoride release from filling materials only results in superficially increased remineralisation of underlying demineralised dentin. PMID- 16251798 TI - Fluoride dose response in pH-cycling models using bovine enamel. AB - The aim of this study was to establish methodologies for verification of the fluoride solution dose-response relationship using bovine enamel and pH-cycling models. Six models of the cariogenic challenge were performed, varying the time of demineralization and pH, time of remineralization, composition of de- and remineralization solutions, frequency and time of application of treatment solutions and pH-cycling duration. For the evaluation of the fluoride effect on caries dynamics, two proposed models provided for improvement in standardization of methods leading to a higher level of precision, demonstrating a dose response between treatments with regard to surface microhardness and DeltaZ. For the evaluation of the fluoride effect on enamel remineralization, the addition of fluoride to the de- and remineralization solutions and the reduction of frequency and time of application of fluoride solutions led to a more suitable pH-cycling model. PMID- 16251799 TI - Synergistic inhibition by combination of fluoride and xylitol on glycolysis by mutans streptococci and its biochemical mechanism. AB - The purpose of this study was to evaluate the combined inhibitory effect of fluoride and xylitol on acid production by mutans streptococci, Streptococcus mutans NCTC10449 and Streptococcus sobrinus 6715, from glucose under strictly anaerobic conditions at fixed pH 5.5 and 7.0. The bacteria were grown in a tryptone-yeast extract broth under strictly anaerobic conditions (N2: 80%; H2: 10%; CO2: 10%). Reaction mixtures for acid production from glucose contained bacterial cells with fluoride (0-6.4 mM) and/or xylitol (60 mM). Acidic end products of glucose fermentation and intracellular glycolytic intermediates were assayed. The combination of fluoride and xylitol inhibited acid production more effectively than fluoride or xylitol alone. In the presence of fluoride and xylitol, the proportion of lactic acid in the total amount of acidic end products decreased, while the proportion of formic and acetic acids increased. Analyses of intracellular glycolytic intermediates revealed that xylitol inhibited the upper part of the glycolytic pathway, while fluoride inhibited the lower part. This study indicates that fluoride and xylitol together have synergistic inhibitory effects on the acid production of mutans streptococci and suggests that xylitol has the potential to enhance inhibitory effects of low concentrations of fluoride. PMID- 16251800 TI - Approximal caries development in adolescents with low to moderate caries risk after different 3-year school-based supervised fluoride mouth rinsing programmes. AB - The aim was to evaluate a 3-year randomised controlled trial of school-based fluoride mouth rinsing (FMR) on approximal caries development in 13- to 16-year olds with low to moderate caries risk. The adolescents used F toothpaste at home and underwent prophylactic treatment at yearly check-ups at public dental clinics. Out of 788 randomly selected 13-year-olds, 622 completed the trial, carried out in 1999-2003. Supervised by a dental nurse, the subjects rinsed with a 0.2% NaF solution at different intervals. Group 1 rinsed their teeth on the first three schooldays every semester; group 2 on the first three and the last three schooldays every semester; group 3 on three consecutive days once a month during semesters; group 4 once every fortnight during semesters, and group 5 (control) did not rinse. Radiographic recording of approximal caries was performed. FMR on the three first and the three last schooldays every semester (group 2) had a prevented fraction of 59%, with approximal enamel lesions as a diagnostic threshold. Corresponding figures for groups 1, 3 and 4 were 30, 47 and 41%, respectively. The control group differed statistically from groups 2-4 for new enamel and dentin lesions and fillings (p < 0.01). Enamel lesions constituted more than 90% of the new caries lesions. Caries progression was low for all groups and no significant differences were found between groups. The main conclusion from this randomised controlled trial is that school-based FMR, as a supplement to the daily use of F toothpaste, reduces caries incidence on approximal surfaces in adolescents with low to moderate caries risk. PMID- 16251801 TI - Non-invasive fluorescence imaging of cell death in fresh human colon epithelia treated with 5-Fluorouracil, CPT-11 and/or TRAIL. AB - Apoptosis is instrumental in several physiological/pathophysiological processes and is a frequently used end-point in the development of anti-neoplastic compounds. Despite ample data on several colon cancer cell lines, little is known about the susceptibility of human colon to apoptosis following treatment with established chemotherapeutics. By treating fresh human colonic explants with 5 Fluorouracil (200 microg/ml), CPT-11 (100 microg/ml) and/or TRAIL (100 ng/ml) we readily detected a signal in situ using FITC-VAD-FMK at different time points, whereas labeling of colonic explants with EGFP-conjugated Annexin V proved less specific. Although TRAIL treatment alone appeared to cause little apoptosis in human colonic epithelia versus the control, we observed a greater number of cells undergoing apoptosis when a combination of CPT-11 and TRAIL was used as compared to either agent alone. This is the initial demonstration of TRAIL-induced apoptosis with or without a chemotherapeutic agent in fresh primary human colon epithelia explants. Thus, human colonic explants may provide a valuable reference point when candidate therapeutic compounds triggering apoptosis in colon cancer cell lines, xenografts or mouse models are developed. The results support the feasibility of developing non-invasive optical imaging strategies to detect apoptosis through direct visualization of injury to human colonic epithelia in vivo. PMID- 16251802 TI - BRCA2-deficient CAPAN-1 cells are extremely sensitive to the inhibition of Poly (ADP-Ribose) polymerase: an issue of potency. AB - We have previously demonstrated that deficiency of either the BRCA1 or BRCA2 breast cancer susceptibility proteins confers substantial cellular sensitivity to the inhibition of Poly(ADP-Ribose) polymerase (PARP). PARP is a key enzyme in the repair of single strand DNA damage via the Base Excision Repair pathway. We suggested that PARP inhibition produces persistent single-strand DNA breaks or gaps which degenerate into stalled replication forks and double-strand breaks, which may be repaired by homologous recombination, a process partially dependent on BRCA1 and BRCA2. It has recently been suggested that our results might be limited to certain BRCA2 mutations as the CAPAN-1 cell line, which carries a naturally occurring 6174delT mutation in one BRCA2 allele accompanied by loss of the wild-type allele, is apparently insensitive to two PARP inhibitors 3 aminobenzamide (IC50 33 microM) and NU1025 (IC50 400 nM). Here we show that CAPAN 1 cells are in fact very sensitive to the potent PARP inhibitors KU0058684 (IC50 3.2 nM) and KU0058948 (IC50 3.4 nM). In contrast, our results reveal much less sensitivity to a chemically related but much less active compound KU0051529 (IC50 730 nM) and to NU1025. These results confirm that treatment with potent PARP inhibitors remains an exciting potential therapy for cancers involving BRCA1 or BRCA2 deficiency. PMID- 16251804 TI - HIV/hepatitis C virus co-infection: its human face. PMID- 16251803 TI - Whole-genome expression profiling of the melanoma progression pathway reveals marked molecular differences between nevi/melanoma in situ and advanced-stage melanomas. AB - Over the past two decades, several known genes have been shown to govern important functions in the development of primary and metastatic melanomas. However, from this limited number of genes, it is not possible to establish detailed molecular profiles for the early and advanced stages of melanoma development. To gain insights into the genetic profile of every stage of the melanoma progression pathway, and to determine to what extent these profiles are similar or distinct, we performed whole-genome expression profiling of tissue specimens representing normal skin, benign and atypical nevi, and early and advanced-stage melanomas. The results of this study provide first-time evidence that significant molecular changes occur distinctly at the border of/transition from melanoma in situ to primary melanoma, and that genes involved in mitotic cell cycle regulation and cell proliferation constitute the two leading categories of genes associated with these changes. PMID- 16251805 TI - Integrated hepatitis C virus treatment: addressing comorbid substance use disorders and HIV infection. AB - OBJECTIVES: To examine hepatitis C virus (HCV) and HIV testing patterns within the Northwest Veterans Integrated Service Network (VISN 20). METHODS: Using a comprehensive VISN 20 database, we retrospectively reviewed medical records of 293,445 veterans. RESULTS: 32.8% of patients were tested for HCV, 5.5% were tested for HIV, and 4.3% were co-tested. Of those tested, 12.3% were HCV positive, 5.4% were HIV positive, and 1.6% were co-infected. 79.1% of HIV positive patients were tested for HCV, 29.2% of whom tested positive. 34.8% of HCV-positive patients were tested for HIV, 4.9% of whom tested positive. Of those tested, HCV-positive patients were significantly more likely than HCV-negative patients to test positive for HIV; HIV-positive patients were no more likely to test positive for HCV than HIV-negative patients. HIV-positive patients with substance use disorders (SUD) were significantly more likely to test HCV positive than those without. Within the total sample, veterans with SUD were significantly more likely to be tested for both diseases and to test positive for HCV but not HIV. After controlling for other categories of SUD, veterans with a history of cocaine abuse compared with those without were at an increased risk of HIV infection and co-infection. CONCLUSION: 79.1% of HIV-positive but only 34.8% of HCV-positive veterans were co-tested, suggesting barriers to HIV testing may exist in VISN 20. Results also indicate that HCV-positive patients are at increased risk for HIV infection and that HIV-positive patients with SUD are at increased risk of HCV infection; routine co-testing for these patients is therefore warranted. Given significant co-infection rates, HCV and HIV screening and testing should be increasingly integrated. Increased infection rates among patients with SUD also warrant integration of HCV and HIV screening and testing into mental health and addiction programmes. PMID- 16251806 TI - Effects of hepatic function and hepatitis C virus on the nervous system assessment of advanced-stage HIV-infected individuals. AB - OBJECTIVES: To examine the effects of liver function and hepatitis C virus (HCV) serostatus on neurological, neuropsychological, and psychiatric abnormalities in an advanced-stage HIV-infected cohort. DESIGN: A correlational analysis of baseline data accumulated on 137 participants in the Manhattan HIV Brain Bank, a longitudinal study of HIV-infected individuals. METHODS: Patients underwent a battery of neuropsychological tests, a semi-structured psychiatric interview, and a neurological examination. The resulting diagnostic data were correlated with biochemical indices of hepatic function and HCV serostatus. RESULTS: Biochemical indices of liver function correlated with motor dysfunction determined by neurological evaluation, but not with neuropsychological or psychiatric disorders. Discrete neurological diagnostic entities showed no relationship with biochemical indices, with one exception: patients with cryptococcal leptomeningitis had worse liver function than those without. HCV had no relationship with any neurological disorder or symptom complex. In contrast, HCV serostatus was related to neuropsychological and psychiatric abnormalities, and indices of liver function were not. HCV-seropositive patients were more likely to have histories of opiate, cocaine or stimulant dependency, to have greater impairment in executive functioning, and to meet diagnostic criteria for AIDS dementia, compared with HCV-negative individuals of similar immunological and virological status. CONCLUSIONS: HCV and biochemical indices of liver function associate differentially with nervous system abnormalities in this HIV-infected population. Neurological abnormalities correlate with biochemical indices of liver function, whereas neuropsychological and psychiatric dysfunction are linked to HCV infection. We postulate that multifactorial impacts of HCV and liver disease on HIV-related nervous system disorders may originate in different anatomical and cellular compartments. PMID- 16251807 TI - Extrahepatic immunological complications of hepatitis C virus infection. AB - Chronic hepatitis C virus (HCV) infection is frequently associated with a number of extrahepatic complications. In the majority of cases the underlying pathogenetic mechanisms are immune mediated, as evidenced by the presence of circulating autoantibodies (mixed cryoglobulinemia), whereas for others a localized host cellular immune response is implicated (e.g. sialadenitis, lichen planus). In this review, the latest data on the pathogenesis and clinical manifestations of the most common autoimmune extrahepatic manifestations of chronic HCV infection are presented. PMID- 16251808 TI - Hepatitis C virus infection and mixed cryoglobulinaemia vasculitis: a review of neurological complications. AB - Chronic liver disease caused by hepatitis C virus (HCV) infection is commonly associated with extrahepatic manifestations, mainly mixed cryoglobulinaemia. Neurological complications in HCV-infected patients occur predominantly in the peripheral nervous system. Peripheral neuropathy in HCV infection is primarily associated with mixed cryoglobulinaemia. Central nervous system (CNS) involvement is more rarely reported. In this review, peripheral and CNS involvement associated with chronic HCV infection are described. The underlying mechanisms and treatment possibilities are discussed. PMID- 16251809 TI - HIV, hepatitis C and HIV/hepatitis C virus co-infection in vulnerable populations. AB - OBJECTIVE: To describe basic patient demographic and clinical characteristics of HIV-infected and HIV/hepatitis C virus (HCV)-co-infected patients receiving care in the Department of Veterans Affairs (VA) with a focus on some patient factors that place such patients at an increased risk of poor health outcomes. DESIGN: An observational retrospective cohort study. METHODS: The study cohort consisted of veterans in the VA Immunology Case Registry who received care in the VA in 2002. RESULTS: Of 18,349 HIV-infected patients, 6782 (37.0%) were HCV seropositive. Compared with HIV-alone-infected patients, HIV/HCV-co-infected patients were older, more likely to be men, more likely to be black or Hispanic, and more likely to report intravenous drug use as a risk factor for HIV acquisition. HIV/HCV-co-infected patients were more likely to have diagnoses of mental health illness, depression, alcohol abuse, substance abuse and hard drug abuse compared with HIV-alone-infected patients. Co-infected patients were less likely to have a history of an AIDS opportunistic infection ever and were less likely to have received HIV antiretroviral drugs in 2002. CONCLUSION: The VA's HIV and HIV/HCV co-infected patient populations have very high rates of additional comorbid conditions that complicate both the pharmacological therapy and clinical course of both HIV and HCV infections. Given the overlap in viral illness and comorbidities, optimal models of integrated care need to be developed for populations with HIV, HCV, and HIV/HCV co-infection and who need substance abuse treatment or mental healthcare. PMID- 16251810 TI - Peripheral neuropathies associated with HIV and hepatitis C co-infection: a review. AB - Co-infection with HIV and hepatitis C has become increasingly prevalent. It is a major source of morbidity in HIV-infected populations. Distal symmetric polyneuropathy is the most common form of peripheral neuropathy in HIV as well as hepatitis C mono-infection. There is considerable overlap in the symptoms and signs of HIV and hepatitis C neuropathy. It is not known whether there are additive or synergistic effects on the peripheral nerve by these two viruses. There is a need for studies to further elucidate the mechanisms involved. PMID- 16251811 TI - Emerging evidence of hepatitis C virus neuroinvasion. AB - It has been reported that hepatitis C virus (HCV) infection is associated with cognitive dysfunction, fatigue and depression, which do not correlate with the severity of liver disease and cannot be accounted for by hepatic encephalopathy or drug abuse. There is also emerging evidence that HCV infection can have negative neurocognitive effects in HIV-infected cohorts. Magnetic resonance spectroscopy has suggested the likely existence of a biological basis for these effects. HCV replicative forms have recently been detected in autopsy brain tissue and the infected cells have been identified as CD68-positive (macrophages/microglia). These findings raise the possibility that HCV infection of the brain could be directly related to the reported neuropsychological and cognitive changes. HCV is not strictly hepatotropic, as it can also replicate in leukocytes, including monocytes/macrophages. The latter cells could provide access of HCV into the central nervous system ('Trojan horse' mechanism) in a process similar to that postulated for HIV-1. In support of this hypothetical mechanism come reports showing a close relationship between HCV sequences present in the brain and cerebrospinal fluid and sequences found in lymph nodes and peripheral blood mononuclear cells. However, despite some similarities there is a fundamental difference between HIV-1 and HCV infection as the latter does not progress into AIDS-type dementia. PMID- 16251812 TI - Differential display analysis of gene expression in brains from hepatitis C infected patients. AB - OBJECTIVES: Hepatitis C virus (HCV) infection is often associated with cognitive dysfunction, fatigue and depression. The current study was undertaken to determine whether HCV infection affects gene expression in brain tissue. DESIGN: We analysed the gene expression pattern in brain tissue in a group of HCV infected patients compared with HCV-negative controls. METHODS: Brain tissue samples were obtained at autopsy from three HCV-positive patients and three HCV negative control patients. The analysis of gene expression was conducted using differential display and reverse Northern hybridization. Only those genes that were up or downregulated more than 1.8 times were considered to be differentially expressed. RESULTS: Altogether, 29 differentially expressed genes were identified by differential display and subsequently confirmed by reverse Northern hybridization. A prominent finding was the downregulation of mitochondrial oxidative phosphorylation genes in HCV-infected patients. The impairment of brain oxidative/energy metabolism has previously been suggested to be the proximate cause of many disorders that impair mentation. Another finding was the downregulation of some ribosomal protein genes and several genes involved in transcription regulation, perhaps reflecting reduced metabolic activities. CONCLUSION: Our findings suggest for the first time that there may be a biological basis for the neuropsychiatric symptoms and cognitive impairment associated with HCV infection. PMID- 16251813 TI - Hepatitis C virus populations in the plasma, peripheral blood mononuclear cells and cerebrospinal fluid of HIV/hepatitis C virus-co-infected patients. AB - BACKGROUND: Chronic hepatitis C virus (HCV) infection has occasionally been associated with diseases of the central nervous system, thus suggesting that the virus has a direct effect on cerebral function. OBJECTIVE: To investigate the presence and heterogeneity of HCV populations in the cerebrospinal fluid (CSF), plasma and peripheral blood mononuclear cells (PBMC) of HIV-infected and anti-HCV positive patients. METHODS: Reverse transcriptase-polymerase chain reaction was used to detect HCV RNA in 21 paired CSF/plasma samples and 20 PBMC samples from HIV/HCV-positive patients. HCV was genotyped by means of a hybridization assay, and its compartmental heterogeneity was analysed by cloning the related amplicons. RESULTS: All of the plasma samples, 14 out of 20 PBMC samples and five out of 21 CSF samples were positive for HCV RNA. Of the five patients with HCV positive CSF, two had the same genotype in the plasma/CSF/PBMC samples, two had a different genotype in the CSF from that in plasma and PBMC, and one had the same genotype in paired CSF/plasma samples. An analysis of the HCV populations showed that two patients seemingly infected with the same genotype in different compartments harbored mixed infection in the CSF but not in the plasma and PBMC samples. CONCLUSIONS: These findings of different HCV genetic diversification in different compartments suggest that CSF is an independent site of viral replication and persistence. PMID- 16251814 TI - Treatment of hepatitis C virus in HIV patients: a review. AB - BACKGROUND: Survival in HIV infection has improved dramatically in the last decade due to antiretroviral therapy (ART). Due to shared routes of transmission, HCV is found in approximately 30% of HIV infected patients. HCV infection has emerged as a major issue in this population as manifest by a major increase in liver-related mortality. ART-associated hepatotoxicity has been demonstrated to occur more frequently in co-infected individuals and may be severe or fatal in some instances. Thus treatment of HCV has become a priority in HIV infected individuals. OBJECTIVES: The main aims of this review are to summarise and illustrate the current evidence based management of anti-HCV therapy in HIV infected patients. METHODS: A systematic review of the literature was performed using Pubmed and Medline searches. CONCLUSION: All HIV-infected patients should be screened for HCV infection via anti-HCV antibody and HCV RNA polymerase chain reaction. If HCV infection is present, treatment should be considered in those patients with evidence of liver inflammation and fibrosis. Recent studies have demonstrated the safety and efficacy of anti-HCV therapy in HIV-infected individuals with pegylated interferon and ribavirin combination therapy. HCV genotype is predictive of response to therapy and increasing the duration of therapy to 48 weeks has proven to be more effective in patients with genotypes 2 and 3. HCV treatment with interferon based therapy is associated with many unique side effects and toxicities in this population of which the clinician must be aware. PMID- 16251815 TI - Interferon-induced depression and cognitive impairment in hepatitis C virus patients: a 72 week prospective study. AB - OBJECTIVES: This study assessed the rates and course of depressive symptomatology and neurocognitive deficits in hepatitis C virus (HCV) patients undergoing interferon treatment, and explored possible predictors of depression and neurocognitive deficits. DESIGN: In order to obtain objective assessments of depression, and to evaluate cognitive impairment, a 72-week prospective study, comprising 48 weeks of treatment and 24 weeks of post-treatment follow-up was utilized. METHODS: A total of 50 HCV patients were assessed at baseline, and 14 times during pegylated interferon plus ribavirin treatment. Patients were also assessed on four timepoints after the termination of treatment. All patients have previously been treated for hepatitis C infection with interferon and were judged to be treatment resistant in these treatments. Depression was assessed using the Center for Epidemiological Studies Depression (CES-D) questionnaire, and patients were interviewed regarding problems with memory, attention and concentration. RESULTS: Eighty-two per cent of interferon-treated patients developed severe enough depressive symptoms to meet the CES-D criteria for possible major depressive disorder (MDD). Possible MDD onset was most frequent by the first week of treatment, and almost all possible MDD cases were observed by week 8. More severe depressive symptoms at baseline were associated with higher depressive symptoms during interferon treatment. Thirty per cent of patients complained about cognitive problems. In half of these patients cognitive impairments were still reported after the termination of treatment. There was no association between depression during interferon treatment and subjective cognitive complaints. CONCLUSIONS: The findings suggest that depression and cognitive impairments are frequent and persistent side-effects of interferon treatment in treatment-resistant patients. PMID- 16251816 TI - A framework for understanding factors that affect access and utilization of treatment for hepatitis C virus infection among HCV-mono-infected and HIV/HCV-co infected injection drug users. AB - Treatment for hepatitis C virus (HCV) is rarely received by injection drug users (IDU), particularly those co-infected with HIV. We propose a framework for understanding factors that affect utilization and adherence to HCV therapy among HCV mono-infected and HIV/HCV-co-infected IDU. Provision of treatment requires calculation of risks and benefits including evaluation of a number of time varying factors that collectively determine a gradient of treatment eligibility, advisability and acceptability, the relative importance of which may differ in co infected and mono-infected IDU. Treatment eligibility is determined by a number of non-modifiable and modifiable contraindications, the latter of which can change over time rendering patients who were once ineligible eligible. Among those eligible, treatment need can be assessed by liver biopsy and therapy may be deferred in those with no liver disease and started in those with significant liver disease. Among those with moderate disease, further consideration of treatment advisability (medical factors that affect treatment response) and acceptability (individual, provider and environmental barriers) is needed before treatment decisions are made. These factors are dynamic and thus should be continually evaluated even among those who may not initially appear to be ready for treatment. An evaluation of this framework is needed to determine applicability and feasibility. Until then, treatment decisions should be made on an individual basis after careful consideration of these issues by provider and patient and efforts to develop novel strategies for identifying IDU who need treatment most (alternatives to liver biopsy) and multidimensional approaches to deliver treatment for HCV while addressing other factors including HIV infection, depression and drug use should be continued. PMID- 16251817 TI - Hepatitis C virus treatment decision-making in the context of HIV co-infection: the role of medical, behavioral and mental health factors in assessing treatment readiness. AB - Hepatitis C virus (HCV)-related liver disease is among the leading causes of mortality among HIV patients, yet very few co-infected patients receive pegylated interferon and ribavirin combination therapy, the standard of care for chronic HCV. Whereas factors related to the provider, patient and clinic setting all contribute to HCV treatment decision-making, the decision of the provider to recommend or defer treatment is perhaps the most critical determinant of whether a patient receives treatment. This paper reviews the literature related to the medical, behavioral and mental health variables that contribute to providers' assessment of treatment readiness, and associations with treatment response, adherence and retention. A greater understanding of the multilevel factors contributing to HCV treatment decision-making, as well as patient characteristics that predict treatment outcome and adherence, can inform the development of interventions aimed at improving HCV care for HIV patients. PMID- 16251818 TI - HIV/hepatitis C virus co-infection in drug users: risk behavior and prevention. AB - Studies of HIV-positive patients have consistently shown that drug users, in particular injection drug users (IDU), are far more likely to have hepatitis C virus (HCV) infection than other patient groups. HIV incidence and prevalence in IDU has declined in recent years, but HCV remains endemic in this population. HCV antibody prevalence among non-injection users of drugs such as heroin and cocaine is between 5 and 30%, although there are scant data on specific transmission risk behavior. The control of HIV/HCV co-infection must address HCV prevention. Epidemiological studies have suggested that HCV prevalence in IDU is subject to various influences, some of which may be modifiable by interventions. However, studies have not shown consistent effects of various prevention strategies on HCV transmission, including studies of HCV screening and education, drug treatment or needle exchange. Although some large cross-sectional studies in regions where needle exchange is available to a large number of drug injectors have reported declining HCV prevalence, the scale of services needed is a matter of considerable debate and has not been systematically quantified. Priorities for research related to the prevention of HIV/HCV co-infection should include estimating the effect on disease occurrence of eliminating specific risk factors, and specifying the level of resources needed to alter HCV incidence. PMID- 16251819 TI - Reductions in hepatitis C virus and HIV infections among injecting drug users in New York City, 1990-2001. AB - OBJECTIVE: To assess trends in HIV, hepatitis C virus (HCV) and HIV/HCV infection among injecting drug users (IDU) from 1990 to 2001 in New York City. The 1990 2001 time period included a very large expansion of syringe exchange in New York City, from 250,000 to 3,000,000 syringes exchanged annually. METHODS: Cross sectional seroprevalence surveys of IDU entering drug abuse treatment in New York City, with sample sizes for HCV of 72 in 1990-1991 and 412 in 2000-2001. A structured risk behavior questionnaire was administered, and HIV and HCV testing were conducted. HCV testing was performed on de-linked stored serum samples. RESULTS: Over the 1990-2001 period, HIV prevalence declined from 54 to 13%. HCV prevalence declined from 80 to 59% among HIV-seronegative individuals, and from 90 to 63% overall. The estimated HCV incidence in 2000-2001 among new injectors was 18 per 100 person-years at risk. CONCLUSIONS: The large-scale expansion of syringe exchange was temporally associated with large reductions in both HIV and HCV prevalence. The prevalence and incidence of HCV, however, still remain at high levels among IDU in New York City. PMID- 16251820 TI - Provider assessment of eligibility for hepatitis C treatment in HIV-infected homeless and marginally housed persons. AB - PURPOSE: To characterize the group of providers delivering medical care to HIV and hepatitis C (HCV) co-infected homeless and marginally housed individuals in San Francisco and to assess factors affecting provider decisions to initiate HCV treatment in this population. SUBJECTS AND METHODS: The Research in Access to Care for the Homeless (REACH) cohort is a representative sample of HIV-infected homeless and marginally housed individuals identified from single room occupancy hotels, homeless shelters and free lunch programs in San Francisco. Primary care providers (PCP) for active, HIV/HCV co-infected REACH cohort participants were administered face-to-face, semi-structured interviews. REACH participants were administered quarterly face-to-face structured interviews. RESULTS: 52/62 (83.9%) providers were interviewed regarding 133/155 (85.8%) active, HIV/HCV co-infected patients. Providers classified 94/133 (70.7%) patients as ineligible for HCV treatment. The mean number of reasons for ineligibility was 3.2. Most frequent reasons for provider determination of ineligibility included likelihood of poor medication adherence, depression, active injection drug use and patient disinterest in treatment. In addition, structural barriers to treatment included poor access to testing, delays in evaluation by a gastroenterologist and exclusion from treatment of patients with comorbidities. CONCLUSIONS: While HCV infection is common, HCV treatment is rare in the HIV/HCV coinfected urban poor. On average, the PCP in this study are experienced and are familiar with this patient population. There are many reasons for providers classifying patients as ineligible for HCV treatment. While these reasons indicate that treatment is difficult given chaotic lifestyle and concurrent medical conditions of this population, they are not insurmountable barriers. New treatments and strategies are necessary to treat this population with high rates of hepatitis C infection. PMID- 16251821 TI - Supported medical care: a multi-faceted approach to helping HIV/hepatitis C virus co-infected adults with serious mental illness. AB - This paper presents a multi-faceted supported rehabilitation and health management model to inform the development of interventions for mentally ill adults who are HIV/hepatitis C virus co-infected. The model, referred to as 'supported medical care' (SMC), calls for the combination of chronic illness self management skills training and enhanced case management. Illustrative examples of each SMC intervention component are provided. Integrated delivery of these intervention components shows promise for improving the coordination of psychiatric and medical care. Research and related policy challenges regarding ongoing quality improvement efforts are also discussed. PMID- 16251822 TI - Integration of pharmacotherapy for opioid addiction into HIV primary care for HIV/hepatitis C virus-co-infected patients. AB - Pharmacotherapy for substance abuse is a rapidly evolving field comprising both old and new effective treatments for substance use. Opiate agonist therapy has been shown to diminish and often eliminate opiate use. This behavior change has resulted in the reduced transmission of many infections, including HIV, hepatitis C virus (HCV), and an enhanced quality of life. For the past 35 years, the provision of opioid agonist therapy has been limited to opioid treatment programmes. Opioid treatment programmes treat approximately 200,000 of the estimated million opiate-addicted individuals in the United States. With the need to increase the number of treatment opportunities available for opioid-dependent patients, Congress passed the Drug Addiction Treatment Act of 2000, which allows for the treatment of opioid dependence using buprenorphine by a properly licensed physician, including HIV primary care physicians. The integration of buprenorphine treatment for opioid addiction into HIV primary care thus provides a new treatment paradigm to address substance abuse in patients with HIV and HCV infections. PMID- 16251823 TI - Integrating care for patients with infectious, psychiatric, and substance use disorders: concepts and approaches. AB - Patients with chronic viral infections such as HIV/AIDS or hepatitis C often have multiple co-existing problems such as psychiatric and addictive disorders, as well as social problems such as lack of housing, transportation and income that present challenging obstacles to successful management. Because services for these different problems are usually provided by different disciplines in varying locations, fragmentation of care can lead to treatment dropouts, lack of adherence, and poor outcomes. Integration strategies, ranging from simple efforts to improve communication and coordinate care to fully integrated multidisciplinary teams have been used to improve disease management. Although evidence for effectiveness is comprised primarily of observational studies of demonstration programmes, integration may be desirable on a pragmatic basis alone. Quality improvement strategies are attractive vehicles for implementing care integration and measuring its impact. Careful assessment of the problem to be solved and the development of targeted strategies will maximize chances of a successful outcome. PMID- 16251824 TI - Hepatitis C virus and HIV co-infection in people with severe mental illness and substance use disorders. AB - OBJECTIVES: The 5-7% of adults in the United States with severe mental illness (SMI), especially the 50% who are 'dually diagnosed' with co-occurring substance use disorders (SUD), are at an elevated risk of HIV and hepatitis C virus (HCV). However, little is known about HIV/HCV co-infection in this population. This paper examines the prevalence and correlates of HIV, hepatitis C, and HIV/HCV co infection in a large, multisite sample of SMI clients. DESIGN: We conducted a re analysis of data on prevalence and correlates of blood-borne infections in a multisite sample of SMI clients. METHODS: In 1997-1998, 755 SMI clients were tested for HIV, hepatitis B virus and HCV, and assessed for demographic, illness related and other behavioral risk factors for blood-borne infections. The prevalence and correlates of co-infection were examined, as well as the knowledge, attitudes and risk behaviors of individuals with HCV mono-infection. RESULTS: Of the 755 participants, 623 (82.5%) were negative for both HIV and HCV, 23 (3.0%) were positive for HIV, 109 (14.4%) were positive for HCV, and 13 (1.7%) were co-infected with HIV and HCV. Overall, 2.5% of dually diagnosed participants were co-infected, whereas only 0.6% of SMI participants without a comorbid SUD diagnosis were co-infected. Co-infection was associated with psychiatric illness severity, ongoing drug abuse, poverty, homelessness, incarceration, urban residence and minority status. HCV-mono-infected clients continued to engage in high levels of risk behavior for HIV. CONCLUSION: In addition to efforts to identify and treat SMI patients with HIV/HCV co-infection, HCV-mono-infected clients should be targeted for prevention interventions. PMID- 16251825 TI - HIV/hepatitis C virus co-infection: basic, behavioral and clinical research in mental health and drug abuse. PMID- 16251826 TI - Hepatitis C virus infection, substance use and mental illness among homeless youth: a review. AB - OBJECTIVES: Homeless youth are at a high risk of substance abuse, mental illness and blood-borne infections, such as hepatitis C. In this paper, we review the implications of these conditions, discuss the unique challenges faced by homeless youth, and explore potential strategies for harm reduction and intervention in this vulnerable population. RESULTS: Interventions that combine youth-centered, service-based care, street outreach, case management, and motivational interviewing with integrated health services such as hepatitis A/B vaccination, and mental health and substance abuse programmes, are presented as innovative approaches to address the healthcare needs of homeless youth. CONCLUSION: Recommendations for age-appropriate interventions and further research are made. PMID- 16251827 TI - Hepatitis B, hepatitis C, and HIV in correctional populations: a review of epidemiology and prevention. AB - The 2 million persons incarcerated in US prisons and jails are disproportionately affected by hepatitis B virus (HBV), hepatitis C virus (HCV) and HIV, with prevalences of infection two to ten times higher than in the general population. Infections are largely due to sex- and drug-related risk behaviors practised outside the correctional setting, although transmission of these infections has also been documented inside jails and prisons. Public health strategies to prevent morbidity and mortality from these infections should include hepatitis B vaccination, HCV and HIV testing and counseling, medical management of infected persons, and substance abuse treatment in incarcerated populations. PMID- 16251828 TI - Health disparities among African-American and Hispanic drug injectors--HIV, AIDS, hepatitis B virus and hepatitis C virus: a review. AB - Disparities in healthcare access, medical outcomes, and specific chronic diseases have been documented for African-American and Hispanic individuals in comparison with non-Hispanic whites. What may be less well known are those health disparities related to common blood-borne pathogens such as hepatitis B virus (HBV) and hepatitis C virus (HCV). Several studies have shown that African American and Hispanic injection drug users (IDU) have higher prevalence rates of these blood-borne pathogens, in addition to higher prevalence rates of HIV infection and AIDS cases. These blood-borne pathogens may contribute to the increased morbidity and mortality among African-American and Hispanic IDU, and perhaps also that of their sexual partners. This article reviews some of what is currently known about the epidemiology of HIV, AIDS, HBV, and HCV among African American and Hispanic individuals, in general, and IDU in particular. In order to reduce or eliminate these health disparities a comprehensive approach is required that includes case finding, pre and post-test counseling, clinical treatment and management, and community-based behavioral or structural interventions. PMID- 16251829 TI - A review of cognitive impairment and cerebral metabolite abnormalities in patients with hepatitis C infection. AB - Numerous studies have reported associations between chronic hepatitis C virus (HCV) infection and fatigue, depression and impairments in health-related quality of life, which are independent of the severity of liver disease. Although there are a large number of potential explanations for these symptoms, including a history of substance abuse and associated personality types, or the effect of the diagnosis of HCV infection itself, there has been recent interest in the possibility of a biological effect of HCV infection on cerebral function. There is emerging evidence of mild, but significant neurocognitive impairment in HCV infection, which cannot be wholly attributed to substance abuse, co-existent depression or hepatic encephalopathy. Impairments are predominantly in the domains of attention, concentration and information processing speed. Furthermore, in-vivo cerebral magnetic resonance spectroscopy studies in patients with hepatitis C and normal liver function have reported elevations in cerebral choline-containing compounds and reductions in N-acetyl aspartate, suggesting that a biological mechanism may underlie the cognitive findings. The recent detection of HCV genetic sequences in post-mortem brain tissue raises the intriguing possibility that HCV infection of the central nervous system may be related to the reported neuropsychological symptoms and cognitive impairment. PMID- 16251830 TI - The neuropsychological and neurological impact of hepatitis C virus co-infection in HIV-infected subjects. AB - OBJECTIVE: To evaluate the effect of hepatitis C virus (HCV)/HIV co-infection on neuropsychological performance and neurological status in HIV/HCV treatment-naive HIV-1-infected individuals we conducted a cross-sectional study using baseline data from an HIV therapy trial. METHODS: HCV status was determined by the presence of anti-HCV antibodies. Neuropsychological function was evaluated by Trailmaking tests, and the Digit Symbol Task. Depression was assessed using the Center for Epidemiologic Studies--Depression Scale. Sleep quality was evaluated by the Pittsburgh Sleep Quality Index and anxiety by the State-Trait Anxiety Inventory for Adults. A questionnaire was designed grading the severity of a variety of symptoms. RESULTS: Of 264 patients with HCV status data, 30 were HCV positive and 234 were HCV negative. Both groups were comparable except that HCV positive individuals had a higher prevalence of intravenous drug use and lower educational level. The HCV-positive group had a significantly lower neuropsychological performance overall. Multivariate modeling supported an association between HCV infection status with test performance in the Digit Symbol Task and mood parameters even when controlling for potentially confounding variables. Marginal differences were noted with respect to symptom questionnaire scores and global sleep. No differences were noted with respect to anxiety. CONCLUSION: The findings suggest that HCV/HIV co-infection has an adverse impact on neuropsychological function. HCV may also be associated with depressed mood, particularly somatic depressive symptoms. Although confounding contributors to neuropsychological performance are difficult to exclude, exploratory modeling supports the association between HCV infection status and some impairment of neuropsychological performance and depressed mood. PMID- 16251831 TI - The effects of hepatitis C, HIV, and methamphetamine dependence on neuropsychological performance: biological correlates of disease. AB - OBJECTIVE: To determine the effects of hepatitis C virus (HCV) infection on neuropsychological (NP) performance. DESIGN: Cross-sectional analysis of a prospectively enrolled cohort. METHODS: A total of 239 HIV-seropositive and 287 HIV-seronegative subjects enrolled in prospective cohort studies at a single center. Subjects underwent standardized assessments, including comprehensive neuropsychological testing, substance use inventory neuromedical examination, venipuncture, and lumbar puncture. HCV antibody was measured in serum. In seropositive individuals, HCV RNA was measured in plasma and cerebrospinal fluid (CSF). RESULTS: HCV-seropositive subjects performed worse on neuropsychological testing and were almost twice as likely to be diagnosed as globally impaired, compared with those who were HCV seronegative. In a multivariate analysis, HCV, HIV, and methamphetamine dependence were independently associated with worse performance, even after adjusting for Centers for Disease Control stage and antiretroviral use. HCV-RNA levels in plasma were higher in those with memory, but not global, impairment. In cerebrospinal fluid, HCV RNA was below 100 copies/ml in all specimens. In HIV-infected subjects, HCV was associated with higher levels of HIV RNA in CSF, but not in plasma. HCV was also associated with higher levels of monocyte chemotactic protein 1, TNF-alpha, and soluble TNF receptor II. HCV-seropositive subjects did not appear to have advanced liver disease. CONCLUSIONS: HIV, HCV, and methamphetamine independently injure the central nervous system, leading to global neuropsychological impairment. HCV may injure the brain by viral or immune-mediated mechanisms. HCV-associated brain injury may be preventable or reversible because HCV infection is potentially curable. PMID- 16251832 TI - Neuropsychological test performance in patients co-infected with hepatitis C virus and HIV. AB - OBJECTIVE: To determine the effect of co-infection on neuropsychological performance in relatively healthy hepatitis C virus (HCV)-alone patients when compared with HCV/HIV-co-infected patients. DESIGN: To test whether the burden of co-infection with HCV and HIV on the central nervous system results in increased cognitive deficits, we tested 47 HCV-alone and 29 HCV/HIV-co-infected patients on a neuropsychological screening battery of tests of attention, concentration and psychomotor speed. METHODS: The neuropsychological test performance of HCV-alone and HCV/HIV-co-infected patients was compared with normative samples. The test performance between HCV-alone and HCV/HIV-co-infected patients was also assessed. Patients with chronic liver disease were divided on the basis of disease severity as determined by fibrosis stage, according to the METAVIR system. Neuropsychological test performance was correlated with fibrosis stage. RESULTS: As previously reported, HCV patients independent of co-infection status demonstrated deficits on neuropsychological measures of attention, concentration and psychomotor speed. No significant differences were found between patients with HCV-alone and HCV/HIV-co-infected patients on the neuropsychological measures. There was a relationship between neuropsychological test performance and fibrosis stage. CONCLUSION: Relatively healthy patients with HCV (either alone or when co-infected with HIV) may have deficits in the domains of attention, concentration and psychomotor speed. In this study no significant differences were found between patients with HCV alone and HCV/HIV-co-infected patients on neuropsychological measures, but as previously demonstrated, greater fibrosis was associated with poorer performance. PMID- 16251833 TI - Perspectives on HIV/hepatitis C virus co-infection, illicit drug use and mental illness. PMID- 16251834 TI - Relative impact of fatigue and subclinical cognitive brain dysfunction on health related quality of life in chronic hepatitis C infection. AB - OBJECTIVES: To assess the relative impact of fatigue and subclinical cognitive brain dysfunction on the impairment of health-related quality of life (HRQL) in hepatitis C virus (HCV) infection. DESIGN AND METHODS: We performed a cross sectional study in 120 patients with untreated chronic HCV infection to test the hypothesis that the severity of fatigue had an independent effect on HCV associated impairment of HRQL. Patients were investigated using the short-form-36 questionnaire, the fatigue impact scale, the brief fatigue inventory, and P300 event-related potentials, as an objective correlate of neurocognitive function. Patients with decompensated cirrhosis or clinical depression were excluded. RESULTS: Relative to healthy controls, HCV-infected patients showed significant levels of fatigue (Fatigue Impact Scale, 49 versus 26 points, brief fatigue inventory, 3.0 versus 1.6 points, P < 0.001). Fatigue impact scale and brief fatigue inventory scores were highly correlated (r = 0.77, P < 0.001), demonstrating concurrent validity. Severity of fatigue and age were the only factors independently associated with the impairment of HRQL (P < 0.001). Fatigue was not related to the severity of hepatitis or the degree of subclinical brain dysfunction. CONCLUSION: In untreated patients with chronic HCV infection, fatigue severity and age but not neurocognitive dysfunction or hepatic function are independently associated with impaired HRQL. Both the fatigue impact scale and the brief fatigue inventory are suitable tools to assess the subjective burden of fatigue. Our findings stress the need for effective therapeutic interventions to reduce the burden of fatigue in patients with HCV infection. PMID- 16251835 TI - Neurological and neuropsychiatric syndromes associated with liver disease. AB - The clinical presentation of acute liver failure and hepatic encephalopathy (HE) in patients with cirrhosis differs significantly. The most serious neurological complication of acute liver failure is the development of devastating brain oedema. Therefore, intracranial pressure monitoring is urgently needed in these patients. Brain oedema is amplified by hypoglycemia, hypoxia and seizures, which are also frequent complications of acute liver failure. Therefore, these parameters must also be monitored. In contrast to acute liver failure in which cerebral dysfunction progresses rapidly, cognitive decline may be clinically undetectable for a long time in cirrhotic patients, until clinically overt symptoms such as psychomotor slowing, disorientation, confusion, extrapyramidal and cerebellar symptoms or a decrease in consciousness occur. Clinically, overt HE is preceded by minimal alterations of cerebral function that can only be detected by neuropsychological or neurophysiological measures, but which nevertheless interfere with the patient's daily living. Rapidly progressing spastic paraparesis (hepatic myelopathy) is a rare complication of cirrhosis. In contrast to HE, it does not respond to blood ammonia lowering therapies but must be considered as an indication for urgent liver transplantation. Cognitive dysfunction has recently been detected in hepatitis C virus (HCV)-infected patients with normal liver function. The patients presented with severe fatigue, cognitive dysfunction and mood disorders. Alterations in brain metabolites, as detected by magnetic resonance spectroscopy, indicated central nervous system alteration in these patients. In contrast to patients with HE, HCV-infected patients did not show motor symptoms or deficits in visual perception, but considerable deficits in attention and concentration ability. PMID- 16251836 TI - Relative prevalence of comorbidities and treatment contraindications in HIV-mono infected and HIV/HCV-co-infected veterans. AB - OBJECTIVES: To determine the prevalence of hepatitis C virus (HCV) co-infection among HIV-infected veterans, assess the prevalence of comorbid conditions that may complicate or limit treatment options, and ascertain whether comorbid conditions were more common in co-infected veterans. DESIGN AND METHODS: We used the Veterans Administration electronic medical records system to identify all veterans receiving care for HIV during fiscal years 1997-2002. Demographic data and diagnostic codes for HIV, HCV, and comorbid conditions were extracted. The validity of using diagnostic codes was assessed by calculating the agreement between chart extraction and electronic data on a separate sample of veterans. Factor analysis was used to identify the structure underlying the intercorrelation between comorbid conditions. Logistic regression was used to compare the prevalence of comorbid conditions and factors between HIV/HCV-co infected and HIV-mono-infected veterans, adjusting for age and race. RESULTS: We identified 25,116 HIV-infected veterans in care, of whom 4489 (18%) were HCV co infected. A validity assessment revealed moderate agreement between chart extraction and electronic data for each of the comorbid conditions assessed. HIV/HCV-co-infected veterans were significantly more likely to have each of the comorbid conditions, and to have significantly more comorbid conditions. Factor analysis revealed three dimensions of comorbidity: mental disorders, medical disorders, and alcohol-related complications. Veterans with co-infection were significantly more likely to have mental disorders and alcohol-related complications. CONCLUSIONS: HIV/HCV-co-infected veterans had a higher prevalence of comorbid conditions that may complicate and limit treatment options for HIV and for HCV co-infection. Strategies to improve treatment options for co-infected patients with comorbidities must be developed. PMID- 16251843 TI - Renal artery stenosis. Introduction. PMID- 16251838 TI - Spinal cord injury clinical trials for neurologic restoration: improving care through clinical research. PMID- 16251844 TI - Medical treatment of renal artery stenosis: is it effective and appropriate? AB - Recent advances in endovascular technology have radically changed the options available for the clinical management of the patient with renovascular disease. These treatment options have fueled an ongoing debate concerning the appropriateness of interventional endovascular therapy for the stenotic renal artery versus conservative medical management. This review examines a typical clinical case scenario and analyzes relevant published literature and the recent guidelines from the Seventh Report of the Joint National Committee on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure (JNC 7) highlighting the significant shortcomings of evidence-based data when it comes to the management of this complex patient population. Early diagnosis provides the best opportunity for appropriate utilization of therapeutic options and rational timing of deployment of interventional techniques. Recommendations for conservative medical management are made based on the review of the medical management arms of the published interventional series. In addition, suggestions are made for practical modifications to the JNC 7 hypertension management protocol to better address the challenging diagnostic and management issues raised by the renovascular patient. PMID- 16251845 TI - Intervention for renal artery stenosis: endovascular and surgical roles. AB - The treatment options for renal artery stenosis include bypass surgery, surgical endarterectomy, or balloon angioplasty with/without stenting. Each of these procedures is delivered today with differing frequency, morbidity/mortality, and outcomes. The procedure most applicable to patients with atherosclerotic disease is percutaneous transluminal renal angioplasty with stenting. Stents prevent plaque recoil, minimizing early restenosis, and the relatively large size of the renal artery (5-7 mm) minimizes late stent restenosis rates. The clinical features that help predict a favorable response to intervention are reviewed. In short, intervention provides a durable means to control renovascular hypertension, ischemic nephropathy, and congestive heart failure due to poor renal volume control. PMID- 16251846 TI - Renal artery stenosis. Conclusions. PMID- 16251847 TI - Atherosclerotic renal artery stenosis: how big is the problem, and what happens if nothing is done? AB - Renal artery stenosis is a common problem, particularly for patients with other manifestations of atherosclerosis. Wide practice variations are apparent regarding how best to manage this disorder. Part of this variation is based on a broad range of clinical presentation, from incidentally identified disease of no clinical importance to rapidly progressive hypertension, renal failure, and refractory congestive heart failure. Advances in antihypertensive therapy, particularly as a result of angiotensin-converting enzyme inhibition and angiotensin receptor blockade, have led to improved blood pressure control and delayed recognition of renal artery disease. As a result, patients now sent for revascularization are older than before and have high comorbid disease risk, primarily related to cardiovascular events. Clinicians need to be vigilant for evidence of unsuspected renal artery stenosis as a cause of treatment-resistant hypertension and/or renal failure. Renal revascularization should be considered in viable individuals before the development of advanced renal insufficiency. PMID- 16251850 TI - A few memories from the beginning... PMID- 16251851 TI - Mechanisms of action of mycophenolate mofetil in preventing acute and chronic allograft rejection. AB - Mycophenolate mofetil (MMF), a prodrug of mycophenolic acid (MPA), an inhibitor of inosine-5'-monophosphate dehydrogenase, has several immunosuppressant actions. MPA depletes guanosine and deoxyguanosine nucleotides preferentially in T and B lymphocytes, inhibiting proliferation and suppressing cell-mediated immune responses and antibody formation, major factors in acute and chronic rejection. MPA also can induce T-lymphocyte apoptosis. MPA suppresses dendritic cell maturation and can induce human monocyte-macrophage cell line differentiation, decreasing the expression of interleukin (IL)-1 and enhancing expression of the IL-1 receptor antagonist. In addition, MPA inhibits adhesion molecule glycosylation and expression and lymphocyte and monocyte recruitment. Activated macrophages produce nitric oxide (NO) and superoxide, which combine to generate tissue-damaging peroxynitrite. MPA depletes tetrahydrobiopterin and decreases NO production by inducible NO synthase without affecting constitutive NO synthase activity. By these mechanisms, MMF exerts anti-inflammatory activity, which could attenuate both acute and chronic rejection. Unlike calcineurin inhibitors, MMF is nonnephrotoxic and does not induce transforming growth factor-beta production, which is fibrogenic. MMF inhibits arterial smooth muscle cell proliferation, a contributor to graft proliferative arteriopathy, and does not increase blood pressure, cholesterol, or triglyceride levels. By decreasing high-density lipoprotein oxidation and macrophage recruitment, MMF also may delay onset/progression of graft atherosclerosis. Thus, MMF may prevent chronic rejection by several mechanisms. MMF activity is synergistic with that of other agents such as valganciclovir for treating cytomegalovirus infection. MMF also has synergistic activity with angiotensin-converting enzyme inhibitors or angiotensin II receptor antagonists in the treatment of some nephropathies in experimental animals. This combination may prevent progression toward end-stage renal disease in humans with chronic allograft, lupus, and diabetic nephropathies. PMID- 16251852 TI - Review of major clinical trials with mycophenolate mofetil in renal transplantation. AB - Mycophenolate mofetil (MMF) was approved for the prevention of acute rejection following renal transplantation based on the results of three groundbreaking, large, clinical trials that demonstrated a significantly reduced risk of acute rejection in patients receiving MMF when compared with those receiving placebo or azathioprine. These three multicenter, prospective, double-blind trials performed at 55 transplant centers on three continents were the largest immunosuppressive drug trials ever attempted and the first prospective, randomized, double-blind trials ever performed in transplantation. These pivotal trials established a foundation for widespread acceptance of MMF in combination with cyclosporine and steroids as a maintenance regimen for renal transplant patients. The findings of these initial trials that led to the approval of MMF for renal transplantation, including long-term follow-up data, will be reviewed in this paper. The expanding scope of major trials of MMF, including trials in pediatric patients, combination regimens with novel induction therapies or other maintenance agents, and trials in special patient populations such as those at high immunological risk or with deteriorating kidney function, will also be discussed. PMID- 16251853 TI - Mycophenolate mofetil in pediatric renal transplantation. AB - Mycophenolate mofetil (MMF) has become a popular immunosuppressive agent in pediatric renal transplantation and has helped to address the unique challenges of transplantation in this population. This paper reviews the early studies that proved MMF's efficacy in reducing the risk of acute renal rejection, as well as its safety and tolerability in comparison to azathioprine. Key conclusions about the pharmacokinetics of MMF from studies of MMF alone and in combination with other immunosuppressive therapies are outlined. The importance of therapeutic drug monitoring (TDM) in this population is also explored in this review. Finally, recent studies showing that newer agents used in combination with MMF can further increase efficacy and reduce the risk of adverse events such as posttransplant lymphoproliferative disease are discussed. PMID- 16251854 TI - The impact of mycophenolate mofetil on long-term outcomes in kidney transplantation. AB - Mycophenolate mofetil (MMF) has been used in kidney and pancreas transplantation for almost 10 years. In the pivotal phase III trials, MMF use was accompanied by a dramatic reduction of rejection rates in kidney transplantation; however, the impact on graft and patient was undetermined. Analyses of the United States Renal Data System and the Scientific Registry of Transplant Recipients databases later provided a valuable measure of the impact of MMF in improving outcomes. In this review, we provide a synopsis of the prospective studies, including but not limited to the pivotal MMF approval trials, and analyses of the national transplant registries relevant to the long-term impact of MMF in kidney transplantation. Indeed, a substantial body of evidence has shown MMF treatment improves patient survival, graft survival, and death-censored graft survival in kidney transplantation. The beneficial effects of MMF have been particularly notable in high-risk recipients such as African Americans. In coming years, these benefits will require reevaluation in the context of the growing use of novel protocols combining MMF with tacrolimus or sirolimus. PMID- 16251855 TI - Toxicity-sparing protocols using mycophenolate mofetil in renal transplantation. AB - The introduction of triple-therapy regimens that include a calcineurin inhibitor (CNI), steroids, and azathioprine greatly reduced the risk of acute rejection in renal transplantation. However, the long-term use of both CNIs and steroids is associated with serious toxicities that ultimately can impact patient/graft survival. Mycophenolate mofetil (MMF), a highly effective immunosuppressant with no known nephrotoxicity, has been shown to provide benefits in preserving long term renal allograft function relative to azathioprine. For these reasons, MMF has become an integral component of toxicity-sparing maintenance regimens that seek to minimize patient exposure to CNIs and steroids. This paper provides an overview of current strategies for reducing the toxicities associated with these agents, which include both withdrawal and avoidance regimens with or without induction therapy. Data are accumulating that toxicity-sparing regimens involving MMF are safe and decrease the risk of side effects that accompany the use of CNIs and steroids. Future studies will determine how to best implement these regimens in the renal transplant population. PMID- 16251856 TI - Review of major clinical trials with mycophenolate mofetil in cardiac transplantation. AB - Over the past 10 years, the addition of mycophenolate mofetil (MMF) to combination immunosuppressive regimens in cardiac transplant patients has resulted in significant outcomes benefits. Randomized trials and other studies have demonstrated that the use of MMF is associated with a decreased risk of rejection and improved survival. This article will provide an overview of these trials, as well as those evaluating MMF in renal-sparing regimens and in pediatric cardiac transplant recipients. In addition, emerging evidence demonstrating that MMF may provide long-term benefits in reducing cardiac allograft vasculopathy and those evaluating the role of MMF therapeutic drug monitoring in cardiac transplant recipients will be discussed. PMID- 16251857 TI - The rationale for and limitations of therapeutic drug monitoring for mycophenolate mofetil in transplantation. AB - The addition of mycophenolate mofetil (MMF) to calcineurin inhibitor-based regimens reduces the incidence of acute rejection after kidney transplantation. The interpatient variability, changes over time of pharmacokinetic parameters, and the potential for drug interactions make the systemic exposure of mycophenolic acid (MPA) unpredictable at a fixed-dose regimen. An increase in plasma concentration of MPA significantly correlates with a decreased likelihood of an acute rejection after kidney or heart transplantation; therefore, a strategy of therapeutic drug monitoring for MMF therapy could improve outcome. Two large randomized, multicenter, prospective trials investigating the added value of therapeutic drug monitoring for MPA, by comparing fixed-dose treatment with concentration-controlled MMF treatment in kidney transplant recipients, are currently ongoing. More data are needed to fully establish the meaning of the reported prognostic value of preoperative inosine monophosphate dehydrogenase (IMPDH) activity, and longitudinal studies monitoring IMPDH activity after transplantation are eagerly awaited. PMID- 16251858 TI - Malignancy after transplantation. AB - As newer immunosuppressive regimens have steadily reduced the incidence of acute rejection and have extended the life expectancy of allograft recipients, posttransplant malignancy has become an important cause of mortality. In fact, it is expected that cancer will surpass cardiovascular complications as the leading cause of death in transplant patients within the next 2 decades. An understanding of the underlying pathobiology and how to minimize cancer risks in transplant recipients are essential. The etiology of posttransplant malignancy is believed to be multifactorial and likely involves impaired immunosurveillance of neoplastic cells as well as depressed antiviral immune activity with a number of common posttransplant malignancies being viral-related. Although calcineurin inhibitors and azathioprine have been linked with posttransplant malignancies, newer agents such as mycophenolate mofetil and sirolimus have not and indeed may have antitumor properties. Long-term data are needed to determine if the use of these agents will ultimately lower the mortality due to malignancy for transplant recipients. PMID- 16251859 TI - Use of mycophenolate mofetil in autoimmune and renal diseases. AB - Progress in understanding the pathogenesis and treatment of rheumatologic and glomerular diseases such as systemic lupus erythematosus and particularly lupus nephritis has been closely linked with the development of newer immunosuppressive agents. With improved patient survival following the institution of cyclophosphamide and corticosteroid therapy, longer-term management issues came to the forefront, especially how to decrease adverse effects of the immunosuppressive regimen. Many of the immunosuppressive regimens used in lupus patients were first established as efficacious and safe through their use in solid organ transplantation. Mycophenolate mofetil (MMF) is now widely used in the field of transplantation. Following anecdotal reports describing benefits of MMF in lupus and lupus nephritis patients, small studies and finally large randomized, controlled trials have established the use of MMF in these patients, particularly those with lupus nephritis. MMF use in other rheumatologic and renal diseases has been evaluated in only smaller studies and very few randomized controlled trials. Nevertheless, many studies currently are ongoing with this immunosuppressive agent. This article will review the published data and the experience of two major New York medical centers with the use of MMF in autoimmune and renal diseases. PMID- 16251862 TI - Import-associated measles outbreak--Indiana, May-June 2005. AB - On May 29, 2005, the Indiana State Department of Health (ISDH) was notified of suspected measles in a female Indiana resident aged 6 years who was hospitalized in Cincinnati, Ohio, where she had been visiting relatives. Serologic analyses performed by the Ohio State Department of Health Laboratory and a private reference laboratory confirmed the diagnosis of measles. The hospital in Cincinnati and the girl's parents told ISDH she had been at a church gathering in northwestern Indiana on May 15 where a fellow attendee had been ill. This fellow attendee was an adolescent girl aged 17 years, an Indiana resident who had not been vaccinated for measles and who had worked during May 4--14 as a missionary in an orphanage and hospital in Bucharest, Romania, where a large measles outbreak was subsequently reported. The teen had returned to the United States with prodromal fever, cough, conjunctivitis, and coryza, traveling on international and domestic commercial airliners on May 14. The next day the teen attended the church gathering along with others who had not been vaccinated because of nonmedical exemptions. Family members recalled that the teen had a rash on May 16; measles was diagnosed retrospectively, and the teen was identified as the index patient. An outbreak investigation was conducted by ISDH and CDC. This report summarizes 1) the results of that investigation, which identified 34 persons with measles, including three who required hospitalization, 2) the measures taken to control and prevent measles transmission, and 3) recommendations to prevent future cases of measles. PMID- 16251860 TI - Mycophenolate mofetil: a decade of clinical experience. PMID- 16251863 TI - Progress toward global eradication of dracunculiasis, January 2004-July 2005. AB - In 1986, an estimated 3.5 million cases of dracunculiasis occurred in 20 countries, and 120 million persons were at risk for the disease. That year, the World Health Assembly adopted a resolution calling for the eradication of dracunculiasis, also known as Guinea worm disease. This report describes the status of the global dracunculiasis eradication program as of July 2005, indicating that, during January-July 2005, a total of 8,191 indigenous cases of dracunculiasis were reported from nine countries, with at least 150 million persons at risk. Despite the substantial reductions in dracunculiasis cases since 1986, eradication of dracunculiasis will require international commitment and ongoing surveillance and intensified interventions at national, state, and local levels. PMID- 16251864 TI - Estimated exposure of adolescents to state-funded anti-tobacco television advertisements--37 states and the District of Columbia, 1999-2003. AB - The majority of persons who become regular smokers begin smoking during adolescence, making this period critical for preventing tobacco use. Evidence suggests that anti-tobacco mass media campaigns that include paid television advertising reduce youth smoking. With development of anti-tobacco programs in all 50 states during the 1990s, spurred by funding from the 1998 Master Settlement Agreement with major cigarette manufacturers, CDC, and other sources, an increasing number of states instituted anti-tobacco media campaigns. This report summarizes trends in median state estimates for the average number of state-funded anti-tobacco television advertisements to which adolescents aged 12 17 years were exposed per month in 37 states and the District of Columbia (DC) during 1999-2003. The findings indicate that the median state estimate of the number of advertisement exposures per month increased from 0.04 in 1999 to 0.80 in 2002 but declined to 0.63 in 2003. The decline in estimated exposure from 2002 to 2003 is consistent with cutbacks in funding for state tobacco-prevention and control programs during this period. Reduced exposure to state-funded anti tobacco advertising might be contributing to the recent lack of substantial change in youth smoking prevalence from 2002 to 2004, which had been declining substantially since 1997. The majority of states need to implement additional measures to ensure that adolescents are adequately exposed to effective paid anti tobacco advertisements as part of tobacco-prevention activities. PMID- 16251865 TI - Prevalence of epilepsy and health-related quality of life and disability among adults with epilepsy--South Carolina, 2003 and 2004. AB - Epilepsy is a common neurologic disorder and poses substantial burdens on physical and mental health. Epilepsy can interfere with social functioning by limiting employment, educational opportunities, and interpersonal relationships and can increase the risk for death. The annual cost of cases of epilepsy in the United States, including direct medical costs and productivity losses, was estimated at $12.5 billion in 1995. Depending on case definitions and populations studied, epilepsy affects an estimated 0.4%-1.0% of the population with a lifetime prevalence of 1.8%-2.6% in certain state populations. This report analyzes data from the 2003 and 2004 South Carolina Behavioral Risk Factor Surveillance System (BRFSS) surveys, which included questions on epilepsy, health related quality of life (HRQOL), and disability. This report summarizes the results of that analysis, which determined that 2.2% of adults in South Carolina had ever been told they had epilepsy, 1.1% had active epilepsy, and both groups reported worse HRQOL and higher prevalence of disability than those who had never had epilepsy. Health-care providers should screen epilepsy patients for cognitive, emotional, and physical health problems that might negatively affect HRQOL. Patients with active epilepsy and recent seizures should be targeted with interventions that will decrease the risk for adverse physical (e.g., injury) and psychosocial (e.g., unemployment) outcomes that accompany continued seizures. PMID- 16251866 TI - Guidelines for identifying and referring persons with fetal alcohol syndrome. AB - Fetal alcohol syndrome (FAS) results from maternal alcohol use during pregnancy and carries lifelong consequences. Early recognition of FAS can result in better outcomes for persons who receive a diagnosis. Although FAS was first identified in 1973, persons with this condition often do not receive a diagnosis. In 2002, Congress directed CDC to update and refine diagnostic and referral criteria for FAS, incorporating recent scientific and clinical evidence. In 2002, CDC convened a scientific working group (SWG) of persons with expertise in FAS research, diagnosis, and treatment to draft criteria for diagnosing FAS. This report summarizes the diagnostic guidelines drafted by the SWG, provides recommendations for when and how to refer a person suspected of having problems related to prenatal alcohol exposure, and assesses existing practices for creating supportive environments that might prevent long-term adverse consequences associated with FAS. The guidelines were created on the basis of a review of scientific evidence, clinical expertise, and the experiences of families affected by FAS regarding the physical and neuropsychologic features of FAS and the medical, educational, and social services needed by persons with FAS and their families. The guidelines are intended to facilitate early identification of persons affected by prenatal exposure to alcohol so they and their families can receive services that enable them to achieve healthy lives and reach their full potential. This report also includes recommendations to enhance identification of and intervention for women at risk for alcohol-exposed pregnancies. Additional data are needed to develop diagnostic criteria for other related disorders (e.g., alcohol-related neurodevelopmental disorder). PMID- 16251867 TI - Health-related quality of life surveillance--United States, 1993-2002. AB - PROBLEM/CONDITION: Population-based surveillance of health-related quality of life (HRQOL) is needed to promote the health and quality of life of U.S. residents and to monitor progress in achieving the two overall Healthy People 2010 goals: 1) increase the quality and years of healthy life and 2) eliminate health disparities. REPORTING PERIOD: This report examines surveillance-based HRQOL data from 1993 through 2002. DESCRIPTION OF SYSTEM: Survey data from a validated set of HRQOL measures (CDC HRQOL-4) were analyzed for 1993-2001 from the Behavioral Risk Factor Surveillance System (BRFSS) surveys for the 50 states and the District of Columbia (DC) and for 2001-2002 from the National Health and Nutrition Examination Survey (NHANES). These measures assessed self-rated health; physically unhealthy days (i.e., the number of days during the preceding 30 days for which physical health, including physical illness and injury, was not good); mentally unhealthy days (i.e., the number of days during the preceding 30 days for which mental health, including stress, depression, and problems with emotions, was not good); and days with activity limitation (i.e., number of days during the preceding 30 days that poor physical or mental health prevented normal daily activities). A summary measure of overall unhealthy days also was computed from the sum of a respondent's physically unhealthy and mentally unhealthy days, with a maximum of 30 days. RESULTS AND INTERPRETATION: During 1993-2001, the mean number of physically unhealthy days, mentally unhealthy days, overall unhealthy days, and activity limitation days was higher after 1997 than before 1997. During 1993-1997, the percentage of respondents with zero overall unhealthy days was stable (51%-53%) but declined to 48% by 2001. The percentage of respondents with >/=14 overall unhealthy days increased from 15%-16% during 1993-1997 to 18% by 2001. Adults increasingly rated their health as fair or poor and decreasingly rated it as excellent or very good. Women, American Indians/Alaska Natives, persons of "other races," separated or divorced persons, unmarried couples, unemployed persons, those unable to work, those with a <$15,000 annual household income, and those with less than a high school education reported worse HRQOL (i.e., physically unhealthy days, mentally unhealthy days, overall unhealthy days, and activity limitation days). Older adults reported more physically unhealthy days and activity limitation days, whereas younger adults reported more mentally unhealthy days. A seasonal pattern was observed in physically unhealthy days and overall unhealthy days. During 1993-2001, BRFSS respondents in 13 states reported increasing physically unhealthy days; respondents in 13 states and DC reported increasing mentally unhealthy days; respondents in Alabama, Connecticut, Maine, New Jersey, New Mexico, North Carolina, and Oregon reported both increasing physically and mentally unhealthy days; and respondents in 16 states and DC reported increasing activity limitation days. During 2001-2002, NHANES respondents with one or more medical conditions (e.g., arthritis or stroke) reported worse HRQOL than those without such conditions, and those with an increasing number of medical conditions reported increasingly worse HRQOL. PUBLIC HEALTH ACTION: Policy makers and researchers should continue to monitor HRQOL and its correlates in the U.S. population. In addition, public health professionals should expand monitoring to populations currently missed by existing surveys, including institutionalized and homeless persons, adolescents, and children. A key aspect is to study and identify the personal and community determinants of HRQOL in prevention research and population studies, to understand how to improve HRQOL, and to reduce HRQOL disparities. In addition, population health assessment professionals should continue to refine and validate HRQOL, functional status, and self-reported health measures. PMID- 16251868 TI - Phase I and pharmacological study of the farnesyltransferase inhibitor tipifarnib (Zarnestra, R115777) in combination with gemcitabine and cisplatin in patients with advanced solid tumours. AB - This phase I trial was designed to determine the safety and maximum tolerated dose (MTD) of tipifarnib in combination with gemcitabine and cisplatin in patients with advanced solid tumours. Furthermore, the pharmacokinetics of each of these agents was evaluated. Patients were treated with tipifarnib b.i.d. on days 1-7 of each 21-day cycle. In addition, gemcitabine was given as a 30-min i.v. infusion on days 1 and 8 and cisplatin as a 3-h i.v. infusion on day 1. An interpatient dose-escalation scheme was used. Pharmacokinetics was determined in plasma and white blood cells. In total, 31 patients were included at five dose levels. Dose-limiting toxicities (DLTs) consisted of thrombocytopenia grade 4, neutropenia grade 4, febrile neutropenia grade 4, electrolyte imbalance grade 3, fatigue grade 3 and decreased hearing grade 2. The MTD was tipifarnib 200 mg b.i.d., gemcitabine 1000 mg m(-2) and cisplatin 75 mg m(-2). Eight patients had a confirmed partial response and 12 patients stable disease. No clinically relevant pharmacokinetic interactions were observed. Tipifarnib can be administered safely at 200 mg b.i.d. in combination with gemcitabine 1000 mg m(-2) and cisplatin 75 mg m(-2). This combination showed evidence of antitumour activity and warrants further evaluation in a phase II setting. PMID- 16251869 TI - Phase II study of concurrent chemoradiotherapy with capecitabine and cisplatin in patients with locally advanced squamous cell carcinoma of the head and neck. AB - We aimed to evaluate the efficacy and safety of concurrent chemoradiotherapy with capecitabine and cisplatin in patients with locally advanced squamous cell carcinoma of the head and neck (SCCHN). In total, 37 patients with stage III or IV SCCHN were enrolled on the study. The chemotherapy consisted of two cycles of intravenous cisplatin of 80 mg m(-2) on day 1 and oral capecitabine 825 mg m(-2) twice daily from day 1 to day 14 at 3-week intervals. The radiotherapy (1.8-2.0 Gy 1 fraction day(-1) to a total dose of 70-70.2 Gy) was delivered to the primary tumour site and neck. The primary tumour sites were as follows: oral cavity (n=6), oropharynx (n=11), hypopharynx (n=8), larynx (n=3), nasopharynx (n=6), and paranasal sinus (n=3). After the chemoradiotherapy, 29 complete responses (78.4%) and 6 partial responses (16.2%) were confirmed. Grade 3 or 4 neutropenia occurred only in two patients, plus grade 3 febrile neutropenia was observed only in one patient. At a median follow-up duration of 19.8 months, the estimated overall survival and progression-free survival rate at 2-year was 76.8 and 57.9%, respectively. Concurrent chemoradiotherapy with capecitabine and cisplatin was found to be well tolerated and effective in patients with locally advanced SCCHN. PMID- 16251870 TI - Successful receptor-mediated radiation therapy of xenografted human midgut carcinoid tumour. AB - Somatostatin receptor (sstr)-mediated radiation therapy is a new therapeutic modality for neuroendocrine (NE) tumours. High expression of sstr in NE tumours leads to tumour-specific uptake of radiolabelled somatostatin analogues and high absorbed doses. In this study, we present the first optimised radiation therapy via sstr using [(177)Lu-DOTA(0)-Tyr(3)]-octreotate given to nude mice xenografted with the human midgut carcinoid GOT1. The tumours in 22 out of 23 animals given therapeutic amounts showed dose-dependent, rapid complete remission. The diagnostic amount (0.5 MBq [(177)Lu-DOTA(0)-Tyr(3)]-octreotate) did not influence tumour growth and was rapidly excreted. In contrast, the therapeutic amount (30 MBq [(177)Lu-DOTA(0)-Tyr(3)]-octreotate) induced rapid tumour regression and entrapment of (177)Lu so that the activity concentration of (177)Lu remained high, 7 and 13 days after injection. The entrapment phenomenon increased the absorbed dose to tumours from 1.6 to 4.0 Gy MBq(-1) and the tumours in animals treated with 30 MBq received 120 Gy. Therapeutic amounts of [(177)Lu-DOTA(0) Tyr(3)]-octreotate rapidly induced apoptosis and gradual development of fibrosis in grafted tumours. In conclusion, human midgut carcinoid xenografts can be cured by receptor-mediated radiation therapy by optimising the uptake of radioligand and taking advantage of the favourable change in biokinetics induced by entrapment of radionuclide in the tumours. PMID- 16251871 TI - A quantitative analysis of lymphatic vessels in human breast cancer, based on LYVE-1 immunoreactivity. AB - This study was undertaken to determine the highly sensitive method for detecting tumour lymphatic vessels in all the fields of each slide (LV), lymphatic microvessel density (LMVD) and lymphatic vessel invasion (LVI) and to compare them with other prognostic parameters using immunohistochemical staining with polyclonal (PCAB) and monoclonal antibodies (MCAB) to the lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1), and the pan-endothelial marker factor VIII in a series of 67 human breast cancers. In all LYVE-1-stained sections, LV (some of which contained red blood cells) were frequently found localised in extralobular stroma, dermis, connective tissue stroma and adjacent to artery and vein, but were rare within the intralobular stroma or the tumour body (3/67 cases) or areas of widespread invasion. In contrast small blood vessels were observed in intra- and extralobular stroma in the factor VIII-stained sections. Quantitation of vessel numbers revealed that LYVE-1/PCAB detected a significantly larger number of LV than either H&E or LYVE-1/MCAB (P<0.0001). LYVE-1/PCAB detected LVI in 25/67 cases (37.3%) and their presence was significantly associated with both lymph node metastasis (chi(2)=4.698, P=0.0248) and unfavourable overall survival (OS) (P=0.0453), while not relapse- free survival (RFS) (P=0.2948). LMVD had no influence for RFS and OS (P=0.4879, P=0.1463, respectively). Our study demonstrates that immunohistochemistry with LYVE-1/PCAB is a highly sensitive method for detecting tumour LV/LVI in breast cancer and LVI is a useful prognostic indicator for lymphatic tumour dissemination. PMID- 16251872 TI - Early neoplastic and metastatic mammary tumours of transgenic mice detected by 5 aminolevulinic acid-stimulated protoporphyrin IX accumulation. AB - A photodynamic technique for human breast cancer detection founded upon the ability of tumour cells to rapidly accumulate the fluorescent product protoporphyrin IX (PpIX) has been applied to transgenic mouse models of mammary tumorigenesis. A major goal of this investigation was to determine whether mouse mammary tumours are reliable models of human disease in terms of PpIX accumulation, for future mechanistic and therapeutic studies. The haeme substrate 5-aminolevulinic acid (5-ALA) (200 mg kg(-1)) was administered to mouse strains that develop mammary tumours of various histological subtypes upon expression of the transgenic oncogenes HRAS, Polyoma Virus middle T antigen, or Simian Virus 40 large T antigen in the mammary gland. Early neoplastic lesions, primary tumours and metastases showed consistent and rapid PpIX accumulation compared to the normal surrounding tissues, as evidenced by red fluorescence (635 nm) when the tumours were directly illuminated with blue light (380-440 nm). Detection of mouse mammary tumours at the stage of ductal carcinoma in situ by red fluorescence emissions suggests that enhanced PpIX synthesis is a good marker for early tumorigenic processes in the mammary gland. We propose the mouse models provide an ideal experimental system for further investigation of the early diagnostic and therapeutic potential of 5-ALA-stimulated PpIX accumulation in human breast cancer patients. PMID- 16251873 TI - Engineering T cells for cancer therapy. AB - It is generally accepted that the immune system plays an important role in controlling tumour development. However, the interplay between tumour and immune system is complex, as demonstrated by the fact that tumours can successfully establish and develop despite the presence of T cells in tumour. An improved understanding of how tumours evade T-cell surveillance, coupled with technical developments allowing the culture and manipulation of T cells, has driven the exploration of therapeutic strategies based on the adoptive transfer of tumour specific T cells. The isolation, expansion and re-infusion of large numbers of tumour-specific T cells generated from tumour biopsies has been shown to be feasible. Indeed, impressive clinical responses have been documented in melanoma patients treated with these T cells. These studies and others demonstrate the potential of T cells for the adoptive therapy of cancer. However, the significant technical issues relating to the production of natural tumour-specific T cells suggest that the application of this approach is likely to be limited at the moment. With the advent of retroviral gene transfer technology, it has become possible to efficiently endow T cells with antigen-specific receptors. Using this strategy, it is potentially possible to generate large numbers of tumour reactive T cells rapidly. This review summarises the current gene therapy approaches in relation to the development of adoptive T-cell-based cancer treatments, as these methods now head towards testing in the clinical trial setting. PMID- 16251874 TI - Microarray comparative genomic hybridisation analysis of intraocular uveal melanomas identifies distinctive imbalances associated with loss of chromosome 3. AB - Defining regions of genomic imbalance can identify genes involved in tumour development. Conventional cytogenetics has identified several nonrandom copy number alterations (CNA) in uveal melanomas (UVM), which include monosomy 3, chromosome 6 abnormalities and gain of 8q. To gain further insight into the CNAs and define the regions involved more precisely we analysed 18 primary UVMs using 1 Mb BAC microarray comparative genomic hybridisation (CGH). Our analysis showed that the most common genomic imbalances were 8q gain (78%), 6p gain (67%) and monosomy 3 (56%). Two distinct CGH profiles could be delineated on the basis of the chromosome 3 status. The most common genetic changes in monosomy 3 tumours, in our study, were gain of 8q11.21-q24.3, 6p25.1-p21.2, 21q21.2-q21.3 and 21q22.13-q22.3 and loss of 1p36.33-p34.3, 1p31.1-p21.2, 6q16.2-q25.3 and 8p23.3 p11.23. In contrast, disomy 3 tumours showed recurrent gains of only 6p25.3-p22.3 and 8q23.2-q24.3. Our approach allowed definition of the smallest overlapping regions of imbalance, which may be important in the development of UVM. PMID- 16251876 TI - Restoration of TGF-beta signalling reduces tumorigenicity in human lung cancer cells. AB - Members of the transforming growth factor-beta (TGF-beta) family regulate a wide range of biological processes including cell proliferation, migration, differentiation, apoptosis, and extracellular matrix deposition. Resistance to TGF-beta-mediated tumour suppressor function in human lung cancer may occur through the loss of type II receptor (TbetaRII) expression. In this study, we investigated the expression pattern of TbetaRII in human lung cancer tissues by RT-PCR and Western blot analyses. We observed downregulation of TbetaRII in 30 out of 46 NSCLC samples (65%) by semiquantitative RT-PCR. Western blot analyses with tumour lysates showed reduced expression of TbetaRII in 77% cases. We also determined the effect of TbetaRII expression in lung adenocarcinoma cell line (VMRC-LCD) that is not responsive to TGF-beta due to lack of TbetaRII expression. Stable expression of TbetaRII in these cells restored TGF-beta-mediated effects including Smad2/3 and Smad4 complex formation, TGF-beta-responsive reporter gene activation, inhibition of cell proliferation and increased apoptosis. Clones expressing TbetaRII showed reduced colony formation in soft-agarose assay and significantly reduced tumorigenicity in athymic nude mice. Therefore, these results suggest that reestablishment of TGF-beta signalling in TbetaRII null cells by stable expression of TbetaRII can reverse malignant behaviour of cells and loss of TbetaRII expression may be involved in lung tumour progression. PMID- 16251875 TI - Adrenomedullin and tumour angiogenesis. AB - The angiogenic activity of peptide adrenomedullin (AM) was first shown in 1998 . Since then, a number of reports have confirmed the ability of AM to induce the growth and migration of isolated vascular endothelial and smooth muscle cells in vitro and to promote angiogenesis in xenografted tumours in vivo. In addition, knockout murine models point to an essential role for AM in embryonic vasculogenesis and ischaemic revascularisation. AM expression is upregulated by hypoxia (a typical feature of solid tumours) and a potential role as a regulator of carcinogenesis and tumour progression has been proposed based on studies in vitro and in animal models. Nevertheless, translational research on AM, and in particular, confirmation of its importance in the vascularisation of human tumours has lagged behind. In this commentary, we review current progress and potential directions for future research into the role of AM in tumour angiogenesis. PMID- 16251877 TI - Intensive induction chemotherapy with C-BOP/BEP for intermediate- and poor-risk metastatic germ cell tumours (EORTC trial 30948). AB - New chemotherapy regimens are continuously explored in patients with high-risk malignant germ cell tumours (MGCTs). This multicentre phase II trial assessed the efficacy and toxicity of C-BOP/BEP chemotherapy in intermediate and poor prognosis MGCT (IGCCCG criteria). C-BOP/BEP treatment consisted of cycles of cisplatin, vincristine, bleomycin and carboplatin, followed by one cycle of vincristine and bleomycin and three cycles of BEP (bleomycon, etoposide, cisplatin). The trial was designed to demonstrate a 1-year progression-free survival rate of 80%, that is, to exclude a 1-year rate of 70% or less, with a one-sided significance level of 5%. Secondary end points included toxicity, overall survival and the postchemotherapy complete response rate. In total, 16 European hospitals entered 66 eligible patients (intermediate prognosis group: 37; poor prognosis group: 29). A total of 45 patients (68.2%, 95% confidence interval (95% CI): 56.9-79.4%) achieved a complete response (intermediate prognosis: 30; poor prognosis: 15). After a median observation time of 40.4 months (range: 13.7-66.3), the 1-year progression-free survival rate was 81.8% 95% CI: 72.5-91.1%). The 2-year overall survival was 84.5% (95% CI: 75.6-93.3%). In all, 51 patients experienced at least one episode of WHO grade 3/4 leucopenia, and at least one event of grade 3/4 thrombocytopenia occurred in 30 patients. There was no toxic death. With an 82% 1-year progression-free survival and a lower limit of the 95% CI above 70%, the efficacy of C-BOP/BEP is comparable to that of published alternative chemotherapy schedules in high-risk MGCT patients. The treatment's toxicity is manageable in a multicentre setting. In poor prognosis patients, C-BOP/BEP should be compared to standard chemotherapy of four cycles of BEP. PMID- 16251878 TI - Angiogenesis and hypoxia in lymph node metastases is predicted by the angiogenesis and hypoxia in the primary tumour in patients with breast cancer. AB - Hypoxia and angiogenesis are important factors in breast cancer progression. Little is known of hypoxia and angiogenesis in lymph node metastases of breast cancer. The aim of this study was to quantify hypoxia, by hypoxia-induced marker expression levels, and angiogenesis, by endothelial cell proliferation, comparing primary breast tumours and axillary lymph node metastases. Tissue sections of the primary tumour and a lymph node metastasis of 60 patients with breast cancer were immunohistochemically stained for the hypoxia-markers carbonic anhydrase 9 (CA9), hypoxia-inducible factor-1alpha (Hif-1alpha) and DEC-1 and for CD34/Ki-67. Endothelial cell proliferation fraction (ECP%) and tumour cell proliferation fraction (TCP%) were assessed. On haematoxylin-eosin stain, the growth pattern and the presence of a fibrotic focus were assessed. Hypoxia-marker expression, ECP% and TCP% in primary tumours and in lymph node metastases were correlated to each other and to clinico-pathological variables. Median ECP% and TCP% in primary tumours and lymph node metastases were comparable (primary tumours: ECP%=4.02, TCP%=19.54; lymph node metastases: ECP%=5.47, TCP%=21.26). ECP% correlated with TCP% (primary tumours: r=0.63, P<0.001; lymph node metastases: r=0.76, P<0.001). CA9 and Hif-1alpha expression were correlated (primary tumours P=0.005; lymph node metastases P<0.001). In primary tumours, CA9 and Hif-1alpha expression were correlated with DEC-1 expression (P=0.05), presence of a fibrotic focus (P<0.007) and mixed/expansive growth pattern (P<0.001). Primary tumours and lymph node metastases with CA9 or Hif-1alpha expression had a higher ECP% and TCP% (P<0.003); in primary tumours, mixed/expansive growth pattern and fibrotic focus were characterised by higher ECP% (P=0.03). Furthermore, between primary tumours and lymph node metastases a correlation was found for ECP%, TCP%, CA9 and Hif 1alpha expression (ECP% r=0.51, P<0.001; TCP r=0.77, P<0.001; CA9 and Hif-1alpha P<0.001). Our data demonstrate that the growth of breast cancer lymph node metastases is angiogenesis dependent and that angiogenesis and hypoxia in the primary tumour predict angiogenesis and hypoxia in the lymph node metastases. Together with previous findings in breast cancer liver metastases, which grow in 96% of cases angiogenesis independently, these data suggest that both the intrinsic growth characteristics and angiogenic potential of breast cancer cells and the site-specific tumour microenvironment determine angiogenesis and hypoxia in breast cancer. PMID- 16251879 TI - Front-line paclitaxel and irinotecan combination chemotherapy in advanced non small-cell lung cancer: a phase I-II trial. AB - Our purpose was to determine the efficacy of irinotecan plus paclitaxel administered on day 1, repeated every 2 weeks, in untreated patients with advanced or metastatic non-small-cell lung cancer (NSCLC). In total, 56 patients with inoperable or metastatic stage III and IV NSCLC with a histologically or cytologically confirmed diagnosis were enrolled. None of the patients had undergone prior chemotherapy or radiation therapy. Treatment involved irinotecan 125 mg m(-2) and paclitaxel 135 mg m(-2) administered on day 1 and repeated every 2 weeks for a planned number of nine cycles. With a standard dose of paclitaxel at 135 mg m(-2), the dosage of irinotecan was escalated at four levels: 75, 100, 125 and 150 mg m(-2); 125 mg m(-2) was established as the maximum tolerated dose; this dosage was administered to 46 patients. A total of 52 patients (median age 65 years, range 38-77 years) were assessable for toxicity and survival and 46 for response rate. Out of 46 evaluable patients, 19 achieved partial response (41.3%), 17 had stable disease (37%) and 10 (21.7%) experienced disease progression. The median duration of response was 6 months (range 2-9+ months). The main adverse reactions were myelotoxicity (grades 3 and 4) in 10 (19.2%) patients and diarrhoea (grade 3) in four (7.7%) patients. Irinotecan combined with paclitaxel, administered every 2 weeks, appears to be an effective treatment for advanced-stage NSCLC. PMID- 16251880 TI - Galectin-3 immunodetection in follicular thyroid neoplasms: a prospective study on fine-needle aspiration samples. AB - Fine-needle aspiration cytology, which is well established to be accurate for the diagnosis of thyroid cancer, may be inconclusive for the follicular thyroid neoplasms. As galectin-3 was suggested to be a marker of malignant thyrocytes, we investigated whether this protein might be helpful in the diagnosis of aspirates classified as undeterminate by cytology. After establishing an easy processing of aspirates for galectin-3 immunodetection, a series of aspirates categorised as benign (n=63), malignant (n=17) or undeterminate (n=34) was prospectively analysed for galectin-3. Only the patients with malignant or undeterminate lesions underwent surgery. Most lesions (86%) diagnosed as malignant by cytology or after surgery were positive for galectin-3. The majority of lesions (94%) classified as benign by cytology or after surgery was negative for galectin-3. The positive and negative predictive values were 83 and 95%, respectively. When focusing on the undeterminate lesions, the sensitivity and specificity were 75 and 90%, respectively, while the positive and negative predictive values were 82 and 87%, respectively. The specificity and the positive predictive value were higher (100%) when considering the percentage of stained cells. Altogether these results show that galectin-3 constitutes a useful marker in the diagnosis of thyroid lesions classified as undeterminate by conventional cytology. PMID- 16251881 TI - A combination of fibre-rich rye bread and yoghurt containing Lactobacillus GG improves bowel function in women with self-reported constipation. AB - OBJECTIVE: The aim of the study was to investigate the effects of fibre-rich rye bread and yoghurt containing Lactobacillus GG (LGG) on intestinal transit time and bowel function, and to test whether they have an interaction in cases of self reported constipation. DESIGN: The study was carried out as a two-by-two factorial design. SETTING: Free-living subjects. SUBJECTS: A total of 59 healthy women with self-reported constipation, recruited by advertisement. INTERVENTIONS: After a baseline period, the subjects were randomized into four diet groups: (1) rye bread+LGG yoghurt, (2) rye bread, (3) LGG yoghurt, and (4) control. The 3 week dietary intervention was followed by a 3-week follow-up period. During each period, total intestinal transit time was measured and the subjects recorded faecal frequency and consistency, difficulty in defecation and gastrointestinal symptoms. RESULTS: The rye bread shortened total intestinal transit time (mean difference, -0.7; CI(95), -1.1 to -0.2; P=0.007), increased faecal frequency (0.3; CI(95), 0.1 to 0.5; P=0.001), softened faeces (-0.3; CI(95), -0.4 to -0.2; P<0.001) and made defecation easier (-0.4; CI(95), -0.5 to -0.2; P<0.001), but also increased gastrointestinal symptoms (1.6; CI(95), 0.7 to 2.4; P<0.001) compared to the low-fibre toast consumed in the LGG and control groups. There were fewer symptoms in the rye bread+LGG group compared to the rye bread group ( 1.3; CI(95), -2.4 to -0.2; P=0.027). CONCLUSIONS: Fibre-rich rye bread can be recommended in the treatment of constipation, and the simultaneous consumption of LGG yoghurt relieves the adverse gastrointestinal effects associated with increased intake of fibre. SPONSORSHIP: Valio Ltd, R&D, and Fazer Bakeries Ltd. PMID- 16251882 TI - Research goals for folate and related B vitamin in Europe. AB - In the past decade, the understanding of folate bioavailability, metabolism and related health issues has increased, but several problems remain, including the difficulty of delivering the available knowledge to the populations at risk. Owing to the low compliance of taking folic acid supplements, for example, among women of child-bearing age who could lower the risk of having a baby with a neural tube defect, food-based strategies aimed at increasing the intake of folate and other B-group vitamins should be a priority for future research. These should include the development of a combined strategy of supplemental folate (possibly with vitamin B(12)), biofortification using engineered plant-derived foods and micro-organisms and food fortification for increasing folate intakes in the general population. Currently, the most effective population-based strategy to reduce NTDs remains folic acid fortification. However, the possible adverse effect of high intakes of folic acid on neurologic functioning among elderly persons with vitamin B(12) deficiency needs urgent investigation. The results of ongoing randomized controlled studies aimed at reducing the prevalence of hyperhomocysteinemia and related morbidity must be available before food-based total population approaches for treatment of hyperhomocysteinemia can be recommended. Further research is required on quantitative assessment of folate intake and bioavailability, along with a more thorough understanding of physiological, biochemical and genetic processes involved in folate absorption and metabolism. PMID- 16251883 TI - Effect of lifelong iodine supplementation on thyroid 131-I uptake: a decrease in uptake in euthyroid but not hyperthyroid individuals compared to observations 50 years ago. AB - BACKGROUND: In Sweden, iodine has been added to table salt (10 mg/kg) since 1936; this amount was increased in 1966 to 50 mg/kg. OBJECTIVE: To investigate a euthyroid Swedish population (n = 44, 60-65 years) with its entire lifespan with iodine supplementation as for 24-h 131-I uptake (24 h IU) and thyroid nodularity (thyroid scintigraphy). To compare the euthyroid 24 h IU with uptake of thyrotoxic individuals, and with observations from 1955. METHODS: The 24 h IU was used in euthyroid individuals after oral administration of 0.1 MBq/2.7 microCi radioiodine and imaging of the thyroid gland was carried out using 99mTc pertechnetate. RESULTS: In 1999-2000, the mean 24 h IU in the euthyroid individuals was 21% (range 11-33%) and the normal (central 95%) reference interval was 14-30%. Scintigraphy suggested multinodular goitre in three euthyroid individuals. In Graves' patients (n = 53, 50-65 years), the mean 24 h IU was 61% (range 29-89%). In 1955, the 24 h IU in euthyroid individuals was higher (38%, range 10-70%), while hyperthyroid patients had uptake values similar to those recorded in the present investigation (mean 62%, range 40-90%). CONCLUSIONS: The population sample studied had to be small for ethical reasons. We conclude that the reference interval for 24 h IU is 14-30% in this population that had spent its entire lifespan with iodine supplementation. This is lower than that recorded in a Swedish euthyroid population half a century ago having had low-grade table-salt iodine supplementation for 20 years. Values for hyperthyroid patients, however, do not appear to have been affected likewise. PMID- 16251884 TI - Evaluation of laser speckle flowmetry for imaging cortical perfusion in experimental stroke studies: quantitation of perfusion and detection of peri infarct depolarisations. AB - Laser speckle imaging of the exposed cerebral cortex allows detailed examination of the time course and topography of perfusion under different experimental conditions. Here we examine the quantitative capacity of the method and its sensitivity for the detection of peri-infarct depolarisations (PIDs). In four cats anaesthetised with chloralose, the right hemisphere was exposed and the right middle cerebral artery was occluded. The brain was illuminated with a laser diode, the speckle pattern was imaged, and images of inverse speckle correlation time (ICT) were derived from the calculated speckle contrast images. We examined the relationship of ICT with perfusion, as imaged quantitatively using umbelliferone clearance (CBF(umb)). Values of ICT and CBF(umb) were compared and regression parameters were calculated for each experiment. In eight cats, cortical surface direct current (DC) potential was monitored at two locations and detection of PIDs by DC potential and ICT change was compared. ICT- and CBF(umb) derived values of perfusion were closely correlated, with a high degree of significance (P<0.0001). Overall, monitoring of DC potential detected 90% of PIDs, whereas ICT detected 56%. We conclude that (1) laser speckle imaging provides an index of perfusion that has a linear relationship with the clearance rate of umbelliferone within the range of levels of perfusion examined; (2) this relationship is relatively stable between experiments; and (3) the method's ability to detect blood flow changes associated with PIDs likely depends on the noise level of the speckle measurements. PMID- 16251885 TI - Reduction of the cell cycle length by decreasing G1 phase and cell cycle reentry expand neuronal progenitor cells in the subventricular zone of adult rat after stroke. AB - A critical determinant of proliferation of progenitor cells is the duration of the cell division cycle. Stroke increases proliferation of progenitor cells in the subventricular zone (SVZ). Using cumulative and single S-phase labeling with 5-bromo-2'-deoxyuridine, we examined cell cycle kinetics of neural progenitor cells in the SVZ after stroke. In nonstroke rats, 20% of the SVZ cell population was proliferating. However, stroke significantly increased dividing cells up to 31% and these cells had a cell cycle length (T(C)) of 15.3 h, significantly (P < 0.05) shorter than the 19 h Tc in nonstroke SVZ cells. Few terminal deoxynucleotidyl transferase-mediated biotinylated UTP nick end labeling-positive cells were detected in the SVZ cells of nonstroke and stroke groups, suggesting that the majority of dividing cells in the SVZ do not undergo apoptosis. Cell cycle phase analysis revealed that stroke substantially shortened the length of the G1 phase (9.6 h) compared with the G1 phase of 12.6 h in nonstroke SVZ cells (P < 0.03). This reduction in G1 contributes to stroke-induced reduction of T(C) because no significant changes were detected on the length of S, G2 and M phases between two groups. Furthermore, compared with progenitor cells in nonstroke SVZ (10%), a greater proportion (14%) of progenitor cells in stroke SVZ reentered the cell cycle after mitosis (P < 0.05). These results show that an increase in proliferating progenitor cells in the SVZ contributes to stroke-induced neurogenesis and this increase is regulated by shortening the length of the cell cycle, decreasing the G1 phase and increasing cell cycle reentry. PMID- 16251886 TI - ERK1/2 inhibition attenuates cerebral blood flow reduction and abolishes ET(B) and 5-HT(1B) receptor upregulation after subarachnoid hemorrhage in rat. AB - Upregulation of endothelin B (ET(B)) and 5-hydroxytryptamine 1B (5-HT(1B)) receptors via transcription has been found after experimental subarachnoid hemorrhage (SAH), and this is associated with enhanced phosphorylation of the mitogen-activated protein kinase (MAPK) extracellular signal-regulated kinase (ERK1/2). In the present study, we hypothesized that inhibition of ERK1/2 alters the ET(B) and 5-HT(1B) receptor upregulation and at the same time prevents the sustained cerebral blood flow (CBF) reduction associated with SAH. The ERK1/2 inhibitor SB386023-b was injected intracisternally in conjunction with and after the induced SAH in rats. At 2 days after the SAH, cerebral arteries were harvested for quantitative real-time polymerase chain reaction, immunohistochemistry and analysis of contractile responses to endothelin-1 (ET-1; ET(A) and ET(B) receptor agonist) and 5-carboxamidotryptamine (5-CT; 5-HT1 receptor agonist) in a sensitive myograph. To investigate if ERK1/2 inhibition had an influence on the local and global CBF after SAH, an autoradiographic technique was used. At 48 h after induced SAH, global and regional CBF were reduced by 50%. This reduction was prevented by treatment with SB386023-b. The ERK1/2 inhibition also decreased the maximum contraction elicited by application of ET-1 and 5-CT in cerebral arteries compared with SAH. In parallel, ERK1/2 inhibition downregulated ET(B) and 5-HT(1B) receptor messenger ribonucleic acid and protein levels compared with the SAH. Cerebral ischemia after SAH involves vasoconstriction and subsequent reduction in the CBF. The results suggest that ERK1/2 inhibition might be a potential treatment for the prevention of cerebral vasospasm and ischemia associated with SAH. PMID- 16251888 TI - A genome sequencing center in every lab. PMID- 16251889 TI - Association of beta2 adrenergic receptor polymorphisms and related haplotypes with triglyceride and LDL-cholesterol levels. AB - Adrenergic receptors regulate lipid mobilization, energy expenditure and glycogen breakdown. The beta(2) adrenergic receptor (beta(2)-AR) gene may constitute a potential candidate gene to explain part of the genetic predisposition to human obesity and correlated traits. With regard to the association between beta(2)-AR gene polymorphisms and obesity-related metabolic disorders, published reports give conflicting results. We investigated the role of three polymorphisms, and related haplotypes of the beta(2)-AR in the obesity and related traits in a cohort of overweight/obese subjects. We characterized one single nucleotide polymorphism (SNP) in the promoter region (5'LC-Cys19Arg) and two in the coding region (Gly16Arg and Gln27Glu) of the beta(2)-AR in 642 consecutively recruited overweight/obese subjects in whom extensive clinical and biochemical analysis was performed. The effect of the polymorphisms on quantitative variables was investigated using multiple linear regression analysis. 5'LC-Cys19 homozygous showed higher triglyceride and LDL-cholesterol levels compared to 5'LC-Arg19 homozygous (P=0.03 and P=0.01, respectively). Similar increase in triglyceride and LDL-cholesterol levels was observed for Arg/Arg genotype compared to Gly/Gly genotype of Gly16Arg polymorphism (P=0.02 and P=0.01, respectively) and for Gln/Gln genotype compared to Glu/Glu genotype of the Gln27Glu polymorphism (P=0.01 and P=0.03, respectively). The 5'LC-Cys(19)Arg(16)Gln(27) haplotype determined a significant increase in triglyceride and LDL-cholesterol levels compared to 5'LC-Arg(19)Gly(16)Glu(27) haplotype (P=0.05 and P=0.02, respectively). Our findings provide additional weight to previous observations on the influence of these three genetic variants on lipid phenotypes; particularly on the increase of triglycerides and LDL-cholesterol levels in overweight/obese subjects carrying the 5'LC-Cys(19)Arg(16)Gln(27) haplotype. PMID- 16251887 TI - Normoxic resuscitation after cardiac arrest protects against hippocampal oxidative stress, metabolic dysfunction, and neuronal death. AB - Resuscitation and prolonged ventilation using 100% oxygen after cardiac arrest is standard clinical practice despite evidence from animal models indicating that neurologic outcome is improved using normoxic compared with hyperoxic resuscitation. This study tested the hypothesis that normoxic ventilation during the first hour after cardiac arrest in dogs protects against prelethal oxidative stress to proteins, loss of the critical metabolic enzyme pyruvate dehydrogenase complex (PDHC), and minimizes subsequent neuronal death in the hippocampus. Anesthetized beagles underwent 10 mins ventricular fibrillation cardiac arrest, followed by defibrillation and ventilation with either 21% or 100% O2. At 1 h after resuscitation, the ventilator was adjusted to maintain normal blood gas levels in both groups. Brains were perfusion-fixed at 2 h reperfusion and used for immunohistochemical measurements of hippocampal nitrotyrosine, a product of protein oxidation, and the E1alpha subunit of PDHC. In hyperoxic dogs, PDHC immunostaining diminished by approximately 90% compared with sham-operated dogs, while staining in normoxic animals was not significantly different from nonischemic dogs. Protein nitration in the hippocampal neurons of hyperoxic animals was 2-3 times greater than either sham-operated or normoxic resuscitated animals at 2 h reperfusion. Stereologic quantification of neuronal death at 24 h reperfusion showed a 40% reduction using normoxic compared with hyperoxic resuscitation. These results indicate that postischemic hyperoxic ventilation promotes oxidative stress that exacerbates prelethal loss of pyruvate dehydrogenase and delayed hippocampal neuronal cell death. Moreover, these findings indicate the need for clinical trials comparing the effects of different ventilatory oxygen levels on neurologic outcome after cardiac arrest. PMID- 16251890 TI - Stability of BAT26 in tumours of hereditary nonpolyposis colorectal cancer patients with MSH2 intragenic deletion. AB - Colon cancers arising in most patients with hereditary nonpolyposis colorectal cancer (HNPCC) show microsatellite instability (MSI). BAT26, a quasimonomorphic polyA stretch located just 3' of MSH2 exon 5, is considered the most sensitive and specific marker of MSI. A total of 10 HNPCC families with large intragenic MSH2 deletions, encompassing exon 5 and intron 5, identified by multiplex ligation-dependent probe amplification (MLPA) were included in this study. The deletions under study were del1-16, del1-8, del1-7, del1-6, and del3-6, detected in 3, 1, 2, 3, and 1 families, respectively. Although all patients examined from these 10 families developed unstable tumours, 13/19 MSI-H tumours (68 %) surprisingly showed stability of BAT26. By MLPA and MSH2 sequence analyses of the BAT26-stable tumours, we demonstrated that the wild-type MSH2 allele was somatically inactivated by an identical large deletion, with complete loss of intron 5/BAT26 sequences at the tumour DNA level. We could infer that the apparent stability of BAT26 was due to the complete absence of target BAT26 sequences in the tumour sample, which results in exclusive amplification of contaminant normal DNA, containing a single copy of a wild-type stable BAT26 sequence. Identification of a subset of MSH2-related unstable tumours that are not recognized by analysis of BAT26 instability indicates that this marker should never be used alone for rapid MSI screening of HNPCC tumours. Moreover, our findings indicate that BAT26 stability in the context of MSI is strongly suggestive of the presence of a large intragenic MSH2 deletion. PMID- 16251891 TI - Complex disease: pleiotropic gene effects in obesity and type 2 diabetes. PMID- 16251892 TI - On the association of the oxidised LDL receptor 1 (OLR1) gene in patients with acute myocardial infarction or coronary artery disease. AB - The human oxidised low-density lipoprotein receptor 1 (OLR1) gene is a functional candidate for atherosclerosis. An association of the OLR1 gene with acute myocardial infarction (AMI) or coronary artery disease (CAD) has recently been reported. In the present study a total of 677 Italian subjects, 327 CAD-free, 350 CAD, of which 190 with AMI and 160 AMI-free, was genotyped for the following four OLR1 single nucleotide polymorphisms: exon 4 K167N, IVS4 -73C>T, IVS4 -14A>G, and 3'UTR 188 C>T. No statistically significant difference was observed in allele or genotype distribution of the exon 4, intron 4, or 3'UTR SNPs in CAD patients compared to CAD-free subjects, or within CAD, in AMI patients compared to AMI free patients. A correlation was found between the K167N G/G genotype and the increased number of obstructed vessels. Even if the OLR1 genotype frequency distribution data in CAD or AMI subjects here reported do not fully confirm the positive results of some other association studies, an association with a marker of CAD severity was observed. PMID- 16251893 TI - Nonlinear association between CGG repeat number and age of menopause in FMR1 premutation carriers. AB - FMR1 premutations are known to be associated with premature ovarian failure (POF), but the underlying mechanism is unknown. We present evidence for a nonlinear association between menopause age and premutation size suggesting that premutations in the mid-size range are at greatest risk for POF, while larger premutations are at lower risk. PMID- 16251894 TI - Evidence of genetic enrichment for exceptional survival using a family approach: the Leiden Longevity Study. AB - We conducted a sib pair study in very old subjects for the purpose of mapping longevity loci. In the present analysis, we explore whether our recruitment strategy has resulted in a population enriched for a heritable component for exceptional longevity. Our study includes families with at least two long-living siblings (men aged 89 years or above; women aged 91 years or above). Data were collected on date of birth and, if applicable, date of death of parents, brothers and sisters, offspring, and spouses of the long-living participants. Standardised mortality ratios (SMRs) compared with the general Dutch population, were calculated. The SMR for all siblings of the long-living participants was 0.66 (95% CI 0.60-0.73). A similar survival benefit was also observed in the parents (SMR=0.76, 95% CI 0.66-0.87) and in the offspring of the long-living subjects (SMR=0.65, 95% CI 0.51-0.80). The SMR of the spouses of the long-living subjects was 0.95 (95% CI 0.82-1.12). The familial clustering of extended survival is unlikely to be caused by ascertainment bias, because in all analyses the long living participants were excluded. Moreover, it is also unlikely to be caused by environmental factors, because the spouses of the long-living participants had a mortality risk comparable with the general Dutch population, whereas they share the same environment. We conclude that our sample is genetically enriched for extreme survival. PMID- 16251896 TI - Genetics of human heterotaxias. AB - The past decade has seen remarkable advances in defining the molecular mechanisms underlying formation of the embryonic left right (LR) axis. This information is slowly transforming our understanding of human birth defects that are caused by disturbed LR axis patterning. Reversals, isomerisms, or segmental discordances of thoraco-abdominal organ position, that is, classic heterotaxy, clearly indicate embryonic disruption of normal LR patterning. Other isolated birth defects, particularly cardiovascular malformations, may be caused by deficiencies in the same pathways. Here, we review the distinctive clinical features of human heterotaxias and try to summarize the known connections between them and the corresponding developmental pathways. PMID- 16251895 TI - Saethre-Chotzen syndrome caused by TWIST 1 gene mutations: functional differentiation from Muenke coronal synostosis syndrome. AB - The Saethre-Chotzen syndrome (SCS) is an autosomal dominant craniosynostosis syndrome with uni- or bilateral coronal synostosis and mild limb deformities. It is caused by loss-of-function mutations of the TWIST 1 gene. In an attempt to delineate functional features separating SCS from Muenke's syndrome, we screened patients presenting with coronal suture synostosis for mutations in the TWIST 1 gene, and for the Pro250Arg mutation in FGFR3. Within a total of 124 independent pedigrees, 39 (71 patients) were identified to carry 25 different mutations of TWIST 1 including 14 novel mutations, to which six whole gene deletions were added. The 71 patients were compared with 42 subjects from 24 pedigrees carrying the Pro250Arg mutation in FGFR3 and 65 subjects from 61 pedigrees without a detectable mutation. Classical SCS associated with a TWIST 1 mutation could be separated phenotypically from the Muenke phenotype on the basis of the following features: low-set frontal hairline, gross ptosis of eyelids, subnormal ear length, dilated parietal foramina, interdigital webbing, and hallux valgus or broad great toe with bifid distal phalanx. Functional differences were even more important: intracranial hypertension as a consequence of early progressive multisutural fusion was a significant problem in SCS only, while mental delay and sensorineural hearing loss were associated with the Muenke's syndrome. Contrary to previous reports, SCS patients with complete loss of one TWIST allele showed normal mental development. PMID- 16251897 TI - Replication of IGF2-INS-TH*5 haplotype effect on obesity in older men and study of related phenotypes. AB - Interindividual variation of the IGF2-INS-TH region influences risk of a variety of diseases and complex traits. Previous studies identified a haplotype (designated IGF2-INS-TH(*)5 and tagged by allele A of IGF2 ApaI, allele 9 of TH01 and class I alleles of INS VNTR) associated with low body mass index (BMI) in a cohort of UK men. We aimed here both to study whether previous findings relating (*)5 with weight are replicated in a different cohort of men (East Hertfordshire) characterised in more phenotypic detail and to test the effect of this haplotype on related subphenotypes. The PHASE program was used to identify (*)5 and not(*)5 haplotypes. A total of 490 haplotypes were derived from 131 men and 114 women, the frequency of (*)5 being around 9%. Specific tests of (*)5 haplotype (vs not(*)5 haplotypes) conducted included Student's t-test and multiple regression analyses. We observed replication of weight effect for the (*)5 haplotype in men: significant associations with lower BMI (-1.81 kg/m(2), P=0.009), lower waist circumference (-6.3 cm, P=0.001) and lower waist-hip ratio (-5%, P<0.001). This haplotype also marks nearly two-fold lower 120 min insulin (P=0.004) as well as low baseline insulin (-11.02 pmol/l, P=0.043) and low 30 min insulin (-64.44 pmol/l, P=0.072) in a glucose tolerance test. No association between (*)5 and these traits was found in women. Our results, taken together with other data on IGFII levels and TH activity, point to the importance of (*)5 as an integrated polygenic haplotype relevant to obesity and insulin response to glucose in men. PMID- 16251898 TI - Analysing DNA patents in relation with diagnostic genetic testing. AB - In the ongoing debate concerning DNA patents, there is a need for empirical data. We aim at creating this data set for DNA patents related to diagnostic genetic testing. To this end we developed two tools to facilitate this process. First, we set up a search strategy to find the relevant patents. Second, we provide a claim classification template to assist the user in the assessment of the subject matter covered by the patent claims and in creating a comprehensive overview of the patent situation within this field. These tools have been used in a pilot study on 11 selected hereditary disorders. In addition, a detailed analysis of the familial breast and ovarian cancer genes patents retrieved by the developed search strategy and their claim classification, after meticulous reading of the documents, allowed us to better describe the problems which medical geneticists and researchers might face when dealing with the patented technology. PMID- 16251899 TI - Two novel and one known mutation of the TGFBR2 gene in Marfan syndrome not associated with FBN1 gene defects. AB - TGF-beta-receptor 2 (TGFBR2) gene defects have been recently associated with Marfan syndrome (MFS) with prominent cardio-skeletal phenotype in patients with negative fibrillin-1 (FBN1) gene screening. Four mutations have been identified to date in five unrelated families. We screened TGFBR2 gene by direct automated sequencing in two adult patients diagnosed with MFS according to Ghent criteria, and in one girl clinically suspected as affected on the basis of a major cardiovascular criterion and skeletal involvement, all proven not to carry mutations in the exon-intron boundaries of FBN1 gene. We identified two novel and one known TGFBR2 gene mutations in the three unrelated probands. The D446N was identified in a 4-year-old girl with de novo disease characterized by severe cardiovascular disease and skeletal involvement. The M425V and R460H mutations were identified in two familial, autosomal dominant MFSs, both characterized by major cardio-skeletal signs and absence of major ocular signs. The mutation R460H has been recently reported in a family with thoracic aortic aneurysms and dissection. The three mutations are absent in 192 controls and affect evolutionarily conserved residues of the serine/threonine kinase domain (exon 5). Our data support the recently reported association between TGFBR2 gene and MFS without major ocular signs (MFS2). The number of genotyped cases however is too low to confirm that major ocular signs are characteristically absent in MFS2. Accordingly, all patients proven or suspected to be affected by MFS with negative FBN1 gene screening could benefit from rapid investigation of the TGFBR2 gene. PMID- 16251900 TI - Two candidate genes for low platelet count identified in an Asian Indian kindred by genome-wide linkage analysis: glycoprotein IX and thrombopoietin. AB - A genome-wide linkage analysis of platelet count was carried out in a large Asian Indian kindred. Linkage analysis showed one marker (D3S1309) on chromosome 3q with a lod score of 3.26 and another (D3S1282) approximately 30 cM centromeric, with a lod score of 2.52. Multipoint analysis of chromosome 3q identified two peaks with maximum multipoint lod scores of 3.52 and 4.11 under markers D3S1309 and D3S1282, respectively. Two strong candidate genes for platelet variation were identified in the linked region; thrombopoietin (THPO) and glycoprotein IX (GPIX). Resequencing of four individuals revealed five single-nucleotide polymorphisms (SNPs) in THPO and one mutation in the transmembrane region of GPIX. Analysis of variance showed that the GPIX mutation and one THPO SNP accounted for 6 and 4% of the variation in platelet count, respectively. The THPO SNP lies in the 3' untranslated region of the gene and has not been previously reported. The G to A transition at nucleotide 653 resulted in an Ala 156 (GCC) to Thr (ACC) replacement in the GPIX protein. The GPIX mutation was recently identified in a Chinese patient with Bernard-Soulier syndrome (BSS), a rare recessive bleeding disorder characterized by thrombocytopenia and giant platelets. One copy of the GPIX mutation was found in 300 European individuals with platelet counts within the normal range. The results suggest that two QTLs on chromosome 3q influence platelet count variation in the Asian Indian kindred, with the GPIX transmembrane mutation and the 3' UTR SNP in THPO being strong candidates. PMID- 16251901 TI - Haplotype block structure study of the CFTR gene. Most variants are associated with the M470 allele in several European populations. AB - An average of about 1700 CFTR (cystic fibrosis transmembrane conductance regulator) alleles from normal individuals from different European populations were extensively screened for DNA sequence variation. A total of 80 variants were observed: 61 coding SNSs (results already published), 13 noncoding SNSs, three STRs, two short deletions, and one nucleotide insertion. Eight DNA variants were classified as non-CF causing due to their high frequency of occurrence. Through this survey the CFTR has become the most exhaustively studied gene for its coding sequence variability and, though to a lesser extent, for its noncoding sequence variability as well. Interestingly, most variation was associated with the M470 allele, while the V470 allele showed an 'extended haplotype homozygosity' (EHH). These findings make us suggest a role for selection acting either on the M470V itself or through an hitchhiking mechanism involving a second site. The possible ancient origin of the V allele in an 'out of Africa' time frame is discussed. PMID- 16251902 TI - The AID antibody diversification enzyme is regulated by protein kinase A phosphorylation. AB - Antibodies, which are produced by B-lineage cells, consist of immunoglobulin heavy (IgH) and light (IgL) chains that have amino-terminal variable regions and carboxy-terminal constant regions. In response to antigens, B cells undergo two types of genomic alterations to increase antibody diversity. Affinity for antigen can be increased by introduction of point mutations into IgH and IgL variable regions by somatic hypermutation. In addition, antibody effector functions can be altered by changing the expressed IgH constant region exons through IgH class switch recombination (CSR). Somatic hypermutation and CSR both require the B-cell specific activation-induced cytidine deaminase protein (AID), which initiates these reactions through its single-stranded (ss)DNA-specific cytidine deaminase activity. In biochemical assays, replication protein A (RPA), a ssDNA-binding protein, associates with phosphorylated AID from activated B cells and enhances AID activity on transcribed double-stranded (ds)DNA containing somatic hypermutation or CSR target sequences. This AID-RPA association, which requires phosphorylation, may provide a mechanism for allowing AID to access dsDNA targets in activated B cells. Here we show that AID from B cells is phosphorylated on a consensus protein kinase A (PKA) site and that PKA is the physiological AID kinase. Thus, AID from non-lymphoid cells can be functionally phosphorylated by recombinant PKA to allow interaction with RPA and promote deamination of transcribed dsDNA substrates. Moreover, mutation of the major PKA phosphorylation site of AID preserves ssDNA deamination activity, but markedly reduces RPA dependent dsDNA deamination activity and severely impairs the ability of AID to effect CSR in vivo. We conclude that PKA has a critical role in post translational regulation of AID activity in B cells. PMID- 16251908 TI - Is the city safe? PMID- 16251904 TI - Sporting contests: seeing red? Putting sportswear in context. AB - The shirt colour worn by sportsmen can affect the behaviour of the competitors, but Hill and Barton show that it may also influence the outcome of contests. By analysing the results of men's combat sports from the Athens 2004 Olympics, they found that more matches were won by fighters wearing red outfits than by those wearing blue; they suggest that red might confer success because it is a sign of dominance in many animal species and could signal aggression in human contests. Here we use another data set from the 2004 Olympics to show that similar winning biases occur in contests in which neither contestant wears red, indicating that a different mechanism may be responsible for these effects. PMID- 16251909 TI - Waking up to the importance of sleep. PMID- 16251910 TI - Free tips. PMID- 16251913 TI - Trial aims to measure social effects of choosing babies' sex. PMID- 16251912 TI - Migration threatens to send flu south. PMID- 16251915 TI - Europe revamps visa rules to attract world's best minds. PMID- 16251916 TI - Advisers knock Katrina health tests. PMID- 16251918 TI - Graphic detail: twelve years of nuclear traffic. PMID- 16251917 TI - Ministers agree to act on warnings of soaring temperatures in Africa. PMID- 16251920 TI - Neuroscience: while you were sleeping. PMID- 16251921 TI - Physics: far from the frontier. PMID- 16251922 TI - Microbiology: pipe dreams. PMID- 16251923 TI - Patent reform prompts intellectual tug-of-war. PMID- 16251926 TI - Later results don't confirm antidepressant suicide link. PMID- 16251927 TI - Internet forest-watchers a new force for conservation. PMID- 16251928 TI - Noting Croats' difference from other Slavs isn't racist. PMID- 16251929 TI - Call for a cull of pointlessly different reference styles. PMID- 16251930 TI - Deeper into the genome. PMID- 16251936 TI - Exploring life's sweet spot. PMID- 16251937 TI - Genomics: understanding human diversity. PMID- 16251938 TI - Chemistry: a cleaner way to nylon? PMID- 16251939 TI - Solid-state physics: silicon's new shine. PMID- 16251940 TI - Biophysics: helicase snaps back. PMID- 16251941 TI - Materials science: changing face of the chameleon. PMID- 16251943 TI - Cell biology: helices sculpt membrane. PMID- 16251944 TI - Mycology: the whiff of danger. PMID- 16251945 TI - Solid-state physics: spin in the slow lane. PMID- 16251946 TI - Human dynamics: Darwin and Einstein correspondence patterns. AB - In an era when letters were the main means of exchanging scientific ideas and results, Charles Darwin (1809-82) and Albert Einstein (1879-1955) were notably prolific correspondents. But did their patterns of communication differ from those associated with the instant-access e-mail of modern times? Here we show that, although the means have changed, the communication dynamics have not: Darwin's and Einstein's patterns of correspondence and today's electronic exchanges follow the same scaling laws. However, the response times of their surface-mail communication is described by a different scaling exponent from e mail communication, providing evidence for a new class of phenomena in human dynamics. PMID- 16251947 TI - Protein glycosylation: chaperone mutation in Tn syndrome. AB - Tn syndrome is a rare autoimmune disease in which subpopulations of blood cells in all lineages carry an incompletely glycosylated membrane glycoprotein, known as the Tn antigen. This truncated antigen has the sugar N-acetylgalactosamine alpha-linked to either a serine or threonine amino-acid residue, whereas the correct T antigen has an additional terminal galactose; the defect may be due to a malfunction of the glycosylating enzyme T-synthase. Here we show that Tn syndrome is associated with a somatic mutation in Cosmc, a gene on the X chromosome that encodes a molecular 'chaperone' that is required for the proper folding and hence full activity of T-synthase. The production of the autoimmune Tn antigen by a glycosyltransferase enzyme rendered defective by a disabled chaperone may have implications for other Tn-related disorders such as IgA nephropathy, a condition that can result in renal failure. PMID- 16251949 TI - Sleep is of the brain, by the brain and for the brain. AB - Sleep is a widespread biological phenomenon, and its scientific study is proceeding at multiple levels at the same time. Marked progress is being made in answering three fundamental questions: what is sleep, what are its mechanisms and what are its functions? The most salient answers to these questions have resulted from applying new techniques from basic and applied neuroscience research. The study of sleep is also shedding light on our understanding of consciousness, which undergoes alteration in parallel with sleep-induced changes in the brain. PMID- 16251950 TI - Hypothalamic regulation of sleep and circadian rhythms. AB - A series of findings over the past decade has begun to identify the brain circuitry and neurotransmitters that regulate our daily cycles of sleep and wakefulness. The latter depends on a network of cell groups that activate the thalamus and the cerebral cortex. A key switch in the hypothalamus shuts off this arousal system during sleep. Other hypothalamic neurons stabilize the switch, and their absence results in inappropriate switching of behavioural states, such as occurs in narcolepsy. These findings explain how various drugs affect sleep and wakefulness, and provide the basis for a wide range of environmental influences to shape wake-sleep cycles into the optimal pattern for survival. PMID- 16251951 TI - Clues to the functions of mammalian sleep. AB - The functions of mammalian sleep remain unclear. Most theories suggest a role for non-rapid eye movement (NREM) sleep in energy conservation and in nervous system recuperation. Theories of REM sleep have suggested a role for this state in periodic brain activation during sleep, in localized recuperative processes and in emotional regulation. Across mammals, the amount and nature of sleep are correlated with age, body size and ecological variables, such as whether the animals live in a terrestrial or an aquatic environment, their diet and the safety of their sleeping site. Sleep may be an efficient time for the completion of a number of functions, but variations in sleep expression indicate that these functions may differ across species. PMID- 16251952 TI - Sleep-dependent memory consolidation. AB - The concept of 'sleeping on a problem' is familiar to most of us. But with myriad stages of sleep, forms of memory and processes of memory encoding and consolidation, sorting out how sleep contributes to memory has been anything but straightforward. Nevertheless, converging evidence, from the molecular to the phenomenological, leaves little doubt that offline memory reprocessing during sleep is an important component of how our memories are formed and ultimately shaped. PMID- 16251953 TI - Insights from studying human sleep disorders. AB - Problems with sleep are one of the commonest reasons for seeking medical attention. Knowledge gained from basic research into sleep in animals has led to marked advances in the understanding of human sleep, with important diagnostic and therapeutic implications. At the same time, research guided by human sleep disorders is leading to important basic sleep concepts. For example, sleep may not be a global, but rather a local, brain phenomenon. Furthermore, contrary to common assumptions, wakefulness, rapid eye movement (REM) and non-REM sleep are not mutually exclusive states. This striking realization explains a fascinating range of clinical phenomena. PMID- 16251954 TI - What are the memory sources of dreaming? AB - Investigators since Freud have appreciated that memories of the people, places, activities and emotions of daily life are reflected in dreams but are typically so fragmented that their predictability is nil. The mechanisms that translate such memories into dream images remain largely unknown. New research targeting relationships between dreaming, memory and the hippocampus is producing a new theory to explain how, why and when we dream of waking life events. PMID- 16251955 TI - Evolution of indirect reciprocity. AB - Natural selection is conventionally assumed to favour the strong and selfish who maximize their own resources at the expense of others. But many biological systems, and especially human societies, are organized around altruistic, cooperative interactions. How can natural selection promote unselfish behaviour? Various mechanisms have been proposed, and a rich analysis of indirect reciprocity has recently emerged: I help you and somebody else helps me. The evolution of cooperation by indirect reciprocity leads to reputation building, morality judgement and complex social interactions with ever-increasing cognitive demands. PMID- 16251956 TI - Repetitive shuttling of a motor protein on DNA. AB - Many helicases modulate recombination, an essential process that needs to be tightly controlled. Mutations in some human disease helicases cause increased recombination, genome instability and cancer. To elucidate the potential mode of action of these enzymes, here we developed a single-molecule fluorescence assay that can visualize DNA binding and translocation of Escherichia coli Rep, a superfamily 1 DNA helicase homologous to Saccharomyces cerevisiae Srs2. Individual Rep monomers were observed to move on single-stranded (ss)DNA in the 3' to 5' direction using ATP hydrolysis. Strikingly, on hitting a blockade, such as duplex DNA or streptavidin, the protein abruptly snapped back close to its initial position, followed by further cycles of translocation and snapback. This repetitive shuttling is likely to be caused by a blockade-induced protein conformational change that enhances DNA affinity for the protein's secondary DNA binding site, thereby resulting in a transient DNA loop. Repetitive shuttling was also observed on ssDNA bounded by a stalled replication fork and an Okazaki fragment analogue, and the presence of Rep delayed formation of a filament of recombination protein RecA on ssDNA. Thus, the binding of a single Rep monomer to a stalled replication fork can lead to repetitive shuttling along the single stranded region, possibly keeping the DNA clear of toxic recombination intermediates. PMID- 16251957 TI - How Prometheus creates structure in Saturn's F ring. AB - Images of Saturn's narrow and contorted F ring returned by the Cassini spacecraft have revealed phenomena not previously detected in any planetary ring system. The perturbing effect of the inner shepherding satellite, Prometheus, seems to introduce channels through the F ring and a 'streamer'--a line of particles that link the ring to the satellite. The detailed mechanism for the formation of these features has been lacking an explanation. Here we show that these phenomena can be understood in terms of a simple gravitational interaction as Prometheus approaches and recedes from the F ring every 14.7 hours. Our numerical models show that as Prometheus recedes from its closest approach to the F ring, it draws out ring material; one orbital period later, this affected region has undergone keplerian shear and is visible as a channel, in excellent agreement with structures seen in the Cassini images. Prometheus' periodic disruption of the F ring will become more pronounced as the two orbits approach their minimum separation in 2009. The model predicts that the appearance of streamers and the associated channels will vary in a regular fashion on a timescale of one orbital period. PMID- 16251958 TI - Observation of spin Coulomb drag in a two-dimensional electron gas. AB - An electron propagating through a solid carries spin angular momentum in addition to its mass and charge. Of late there has been considerable interest in developing electronic devices based on the transport of spin that offer potential advantages in dissipation, size and speed over charge-based devices. However, these advantages bring with them additional complexity. Because each electron carries a single, fixed value (- e) of charge, the electrical current carried by a gas of electrons is simply proportional to its total momentum. A fundamental consequence is that the charge current is not affected by interactions that conserve total momentum, notably collisions among the electrons themselves. In contrast, the electron's spin along a given spatial direction can take on two values, +/- [planck]/2 (conventionally upward arrow, downward arrow), so that the spin current and momentum need not be proportional. Although the transport of spin polarization is not protected by momentum conservation, it has been widely assumed that, like the charge current, spin current is unaffected by electron electron (e-e) interactions. Here we demonstrate experimentally not only that this assumption is invalid, but also that over a broad range of temperature and electron density, the flow of spin polarization in a two-dimensional gas of electrons is controlled by the rate of e-e collisions. PMID- 16251960 TI - Unidirectional molecular motor on a gold surface. AB - Molecules capable of mimicking the function of a wide range of mechanical devices have been fabricated, with motors that can induce mechanical movement attracting particular attention. Such molecular motors convert light or chemical energy into directional rotary or linear motion, and are usually prepared and operated in solution. But if they are to be used as nanomachines that can do useful work, it seems essential to construct systems that can function on a surface, like a recently reported linear artificial muscle. Surface-mounted rotors have been realized and limited directionality in their motion predicted. Here we demonstrate that a light-driven molecular motor capable of repetitive unidirectional rotation can be mounted on the surface of gold nanoparticles. The motor design uses a chiral helical alkene with an upper half that serves as a propeller and is connected through a carbon-carbon double bond (the rotation axis) to a lower half that serves as a stator. The stator carries two thiol functionalized 'legs', which then bind the entire motor molecule to a gold surface. NMR spectroscopy reveals that two photo-induced cis-trans isomerizations of the central double bond, each followed by a thermal helix inversion to prevent reverse rotation, induce a full and unidirectional 360 degrees rotation of the propeller with respect to the surface-mounted lower half of the system. PMID- 16251959 TI - Strong quantum-confined Stark effect in germanium quantum-well structures on silicon. AB - Silicon is the dominant semiconductor for electronics, but there is now a growing need to integrate such components with optoelectronics for telecommunications and computer interconnections. Silicon-based optical modulators have recently been successfully demonstrated; but because the light modulation mechanisms in silicon are relatively weak, long (for example, several millimetres) devices or sophisticated high-quality-factor resonators have been necessary. Thin quantum well structures made from III-V semiconductors such as GaAs, InP and their alloys exhibit the much stronger quantum-confined Stark effect (QCSE) mechanism, which allows modulator structures with only micrometres of optical path length. Such III-V materials are unfortunately difficult to integrate with silicon electronic devices. Germanium is routinely integrated with silicon in electronics, but previous silicon-germanium structures have also not shown strong modulation effects. Here we report the discovery of the QCSE, at room temperature, in thin germanium quantum-well structures grown on silicon. The QCSE here has strengths comparable to that in III-V materials. Its clarity and strength are particularly surprising because germanium is an indirect gap semiconductor; such semiconductors often display much weaker optical effects than direct gap materials (such as the III-V materials typically used for optoelectronics). This discovery is very promising for small, high-speed, low-power optical output devices fully compatible with silicon electronics manufacture. PMID- 16251961 TI - Low-latitude seasonality of Cretaceous temperatures in warm and cold episodes. AB - The Cretaceous period is generally considered to have been a time of warm climate. Evidence for cooler episodes exists, particularly in the early Cretaceous period, but the timing and significance of these cool episodes are not well constrained. The seasonality of temperatures is important for constraining equator-to-pole temperature gradients and may indicate the presence of polar ice sheets; however, reconstructions of Cretaceous sea surface temperatures are predominantly based on the oxygen isotopic composition of planktonic foraminifera that do not provide information about such intra-annual variations. Here we present intra-shell variations in delta18O values of rudist bivalves (Hippuritoidea) from palaeolatitudes between 8 degrees and 31 degrees N, which record the evolution of the seasonality of Cretaceous sea surface temperatures in detail. We find high maximum temperatures (approximately 35 to 37 degrees C) and relatively low seasonal variability (< 12 degrees C) between 20 degrees and 30 degrees N during the warmer Cretaceous episodes. In contrast, during the cooler episodes our data show seasonal sea surface temperature variability of up to 18 degrees C near 25 degrees N, comparable to the range found today. Such a large seasonal variability is compatible with the existence of polar ice sheets. PMID- 16251962 TI - Cooling of the Earth and core formation after the giant impact. AB - Kelvin calculated the age of the Earth to be about 24 million years by assuming conductive cooling from being fully molten to its current state. Although simplistic, his result is interesting in the context of the dramatic cooling that took place after the putative Moon-forming giant impact, which contributed the final approximately 10 per cent of the Earth's mass. The rate of accretion and core segregation on Earth as deduced from the U-Pb system is much slower than that obtained from Hf-W systematics, and implies substantial accretion after the Moon-forming impact, which occurred 45 +/- 5 Myr after the beginning of the Solar System. Here we propose an explanation for the two timescales. We suggest that the Hf-W timescale reflects the principal phase of core-formation before the giant impact. Crystallization of silicate perovskite in the lower mantle during this phase produced Fe(3+), which was released during the giant impact, and this oxidation resulted in late segregation of sulphur-rich metal into which Pb dissolved readily, setting the younger U-Pb age of the Earth. Separation of the latter metal then occurred 30 +/- 10 Myr after the Moon-forming impact. Over this time span, in surprising agreement with Kelvin's result, the Earth cooled by about 4,000 K in returning from a fully molten to a partially crystalline state. PMID- 16251963 TI - Mammal-like muscles power swimming in a cold-water shark. AB - Effects of temperature on muscle contraction and powering movement are profound, outwardly obvious, and of great consequence to survival. To cope with the effects of environmental temperature fluctuations, endothermic birds and mammals maintain a relatively warm and constant body temperature, whereas most fishes and other vertebrates are ectothermic and conform to their thermal niche, compromising performance at colder temperatures. However, within the fishes the tunas and lamnid sharks deviate from the ectothermic strategy, maintaining elevated core body temperatures that presumably confer physiological advantages for their roles as fast and continuously swimming pelagic predators. Here we show that the salmon shark, a lamnid inhabiting cold, north Pacific waters, has become so specialized for endothermy that its red, aerobic, locomotor muscles, which power continuous swimming, seem mammal-like, functioning only within a markedly elevated temperature range (20-30 degrees C). These muscles are ineffectual if exposed to the cool water temperatures, and when warmed even 10 degrees C above ambient they still produce only 25-50% of the power produced at 26 degrees C. In contrast, the white muscles, powering burst swimming, do not show such a marked thermal dependence and work well across a wide range of temperatures. PMID- 16251964 TI - Reinforcement drives rapid allopatric speciation. AB - Allopatric speciation results from geographic isolation between populations. In the absence of gene flow, reproductive isolation arises gradually and incidentally as a result of mutation, genetic drift and the indirect effects of natural selection driving local adaptation. In contrast, speciation by reinforcement is driven directly by natural selection against maladaptive hybridization. This gives individuals that choose the traits of their own lineage greater fitness, potentially leading to rapid speciation between the lineages. Reinforcing natural selection on a population of one of the lineages in a mosaic contact zone could also result in divergence of the population from the allopatric range of its own lineage outside the zone. Here we test this with molecular data, experimental crosses, field measurements and mate choice experiments in a mosaic contact zone between two lineages of a rainforest frog. We show that reinforcing natural selection has resulted in significant premating isolation of a population in the contact zone not only from the other lineage but also, incidentally, from the closely related main range of its own lineage. Thus we show the potential for reinforcement to drive rapid allopatric speciation. PMID- 16251965 TI - Chimpanzees are indifferent to the welfare of unrelated group members. AB - Humans are an unusually prosocial species-we vote, give blood, recycle, give tithes and punish violators of social norms. Experimental evidence indicates that people willingly incur costs to help strangers in anonymous one-shot interactions, and that altruistic behaviour is motivated, at least in part, by empathy and concern for the welfare of others (hereafter referred to as other regarding preferences). In contrast, cooperative behaviour in non-human primates is mainly limited to kin and reciprocating partners, and is virtually never extended to unfamiliar individuals. Here we present experimental tests of the existence of other-regarding preferences in non-human primates, and show that chimpanzees (Pan troglodytes) do not take advantage of opportunities to deliver benefits to familiar individuals at no material cost to themselves, suggesting that chimpanzee behaviour is not motivated by other-regarding preferences. Chimpanzees are among the primates most likely to demonstrate prosocial behaviours. They participate in a variety of collective activities, including territorial patrols, coalitionary aggression, cooperative hunting, food sharing and joint mate guarding. Consolation of victims of aggression and anecdotal accounts of solicitous treatment of injured individuals suggest that chimpanzees may feel empathy. Chimpanzees sometimes reject exchanges in which they receive less valuable rewards than others, which may be one element of a 'sense of fairness', but there is no evidence that they are averse to interactions in which they benefit more than others. PMID- 16251966 TI - Mapping determinants of human gene expression by regional and genome-wide association. AB - To study the genetic basis of natural variation in gene expression, we previously carried out genome-wide linkage analysis and mapped the determinants of approximately 1,000 expression phenotypes. In the present study, we carried out association analysis with dense sets of single-nucleotide polymorphism (SNP) markers from the International HapMap Project. For 374 phenotypes, the association study was performed with markers only from regions with strong linkage evidence; these regions all mapped close to the expressed gene. For a subset of 27 phenotypes, analysis of genome-wide association was performed with >770,000 markers. The association analysis with markers under the linkage peaks confirmed the linkage results and narrowed the candidate regulatory regions for many phenotypes with strong linkage evidence. The genome-wide association analysis yielded highly significant results that point to the same locations as the genome scans for about 50% of the phenotypes. For one candidate determinant, we carried out functional analyses and confirmed the variation in cis-acting regulatory activity. Our findings suggest that association studies with dense SNP maps will identify susceptibility loci or other determinants for some complex traits or diseases. PMID- 16251967 TI - Wnt signalling regulates adult hippocampal neurogenesis. AB - The generation of new neurons from neural stem cells is restricted to two regions of the adult mammalian central nervous system: the subventricular zone of the lateral ventricle, and the subgranular zone of the hippocampal dentate gyrus. In both regions, signals provided by the microenvironment regulate the maintenance, proliferation and neuronal fate commitment of the local stem cell population. The identity of these signals is largely unknown. Here we show that adult hippocampal stem/progenitor cells (AHPs) express receptors and signalling components for Wnt proteins, which are key regulators of neural stem cell behaviour in embryonic development. We also show that the Wnt/beta-catenin pathway is active and that Wnt3 is expressed in the hippocampal neurogenic niche. Overexpression of Wnt3 is sufficient to increase neurogenesis from AHPs in vitro and in vivo. By contrast, blockade of Wnt signalling reduces neurogenesis from AHPs in vitro and abolishes neurogenesis almost completely in vivo. Our data show that Wnt signalling is a principal regulator of adult hippocampal neurogenesis and provide evidence that Wnt proteins have a role in adult hippocampal function. PMID- 16251968 TI - Regulation of Lethal giant larvae by Dishevelled. AB - The establishment of polarity in many cell types depends on Lgl, the tumour suppressor product of lethal giant larvae, which is involved in basolateral protein targeting. The conserved complex of Par3, Par6 and atypical protein kinase C phosphorylates and inactivates Lgl at the apical surface; however, the signalling mechanisms that coordinate cell polarization in development are not well defined. Here we show that a vertebrate homologue of Lgl associates with Dishevelled, an essential mediator of Wnt signalling, and that Dishevelled regulates the localization of Lgl in Xenopus ectoderm and Drosophila follicular epithelium. We show that both Lgl and Dsh are required for normal apical-basal polarity of Xenopus ectodermal cells. In addition, we show that the Wnt receptor Frizzled 8, but not Frizzled 7, causes Lgl to dissociate from the cortex with the concomitant loss of its activity in vivo. These findings suggest a molecular basis for the regulation of cell polarity by Frizzled and Dishevelled. PMID- 16251969 TI - A heterodimeric complex that promotes the assembly of mammalian 20S proteasomes. AB - The 26S proteasome is a multisubunit protease responsible for regulated proteolysis in eukaryotic cells. It comprises one catalytic 20S proteasome and two axially positioned 19S regulatory complexes. The 20S proteasome is composed of 28 subunits arranged in a cylindrical particle as four heteroheptameric rings, alpha1-7beta1-7beta1-7alpha1-7 (refs 4, 5), but the mechanism responsible for the assembly of such a complex structure remains elusive. Here we report two chaperones, designated proteasome assembling chaperone-1 (PAC1) and PAC2, that are involved in the maturation of mammalian 20S proteasomes. PAC1 and PAC2 associate as heterodimers with proteasome precursors and are degraded after formation of the 20S proteasome is completed. Overexpression of PAC1 or PAC2 accelerates the formation of precursor proteasomes, whereas knockdown by short interfering RNA impairs it, resulting in poor maturation of 20S proteasomes. Furthermore, the PAC complex provides a scaffold for alpha-ring formation and keeps the alpha-rings competent for the subsequent formation of half-proteasomes. Thus, our results identify a mechanism for the correct assembly of 20S proteasomes. PMID- 16251972 TI - Taking addiction research into the clinic. PMID- 16251970 TI - The histone H3.3 chaperone HIRA is essential for chromatin assembly in the male pronucleus. AB - In sexually reproducing animals, a crucial step in zygote formation is the decondensation of the fertilizing sperm nucleus into a DNA replication-competent male pronucleus. Genome-wide nucleosome assembly on paternal DNA implies the replacement of sperm chromosomal proteins, such as protamines, by maternally provided histones. This fundamental process is specifically impaired in sesame (ssm), a unique Drosophila maternal effect mutant that prevents male pronucleus formation. Here we show that ssm is a point mutation in the Hira gene, thus demonstrating that the histone chaperone protein HIRA is required for nucleosome assembly during sperm nucleus decondensation. In vertebrates, HIRA has recently been shown to be critical for a nucleosome assembly pathway independent of DNA synthesis that specifically involves the H3.3 histone variant. We also show that nucleosomes containing H3.3, and not H3, are specifically assembled in paternal Drosophila chromatin before the first round of DNA replication. The exclusive marking of paternal chromosomes with H3.3 represents a primary epigenetic distinction between parental genomes in the zygote, and underlines an important consequence of the critical and highly specialized function of HIRA at fertilization. PMID- 16251974 TI - How neurons keep in touch. PMID- 16251975 TI - Synaptic plasticity and self-organization in the hippocampus. PMID- 16251976 TI - Glial cells under remote control. PMID- 16251977 TI - Senseless makes sense for spinocerebellar ataxia-1. PMID- 16251978 TI - How the brain recovers following damage. PMID- 16251981 TI - The neuroscience of addiction. PMID- 16251982 TI - Neurobiology of addiction: treatment and public policy ramifications. AB - In the United States, efforts to treat addiction are hampered by prejudice and a public view that treats it as a disorder of self-control, not a disease. We highlight select advances in addiction research that, if disseminated to the public, could reverse these misconceptions and facilitate changes in policy to improve treatment access and care delivery for this highly prevalent disease. PMID- 16251983 TI - The role of neuroadaptations in relapse to drug seeking. AB - One of the most difficult problems in treating addiction is not withdrawing addicts from drugs, but preventing relapse. Persistent neuroadaptations are thought to underlie aspects of addiction, including relapse. This commentary assesses the degree to which these neuroadaptations, primarily identified in preclinical studies on cocaine, induce relapse. PMID- 16251984 TI - How do we determine which drug-induced neuroplastic changes are important? AB - Although many drug-induced neural changes are known, progress has been slow in identifying the ones that actually mediate addiction. Identifying changes that are specific to particular elements of the transition from initial to habitual to relapsing drug use may be a fruitful strategy for pinpointing which forms of drug induced plasticity are critical for addiction. PMID- 16251985 TI - Plasticity of reward neurocircuitry and the 'dark side' of drug addiction. AB - Drug seeking is associated with activation of reward neural circuitry. Here we argue that drug addiction also involves a 'dark side'--a decrease in the function of normal reward-related neurocircuitry and persistent recruitment of anti-reward systems. Understanding the neuroplasticity of the dark side of this circuitry is the key to understanding vulnerability to addiction. PMID- 16251986 TI - Is there a common molecular pathway for addiction? AB - Drugs of abuse have very different acute mechanisms of action but converge on the brain's reward pathways by producing a series of common functional effects after both acute and chronic administration. Some similar actions occur for natural rewards as well. Researchers are making progress in understanding the molecular and cellular basis of these common effects. A major goal for future research is to determine whether such common underpinnings of addiction can be exploited for the development of more effective treatments for a wide range of addictive disorders. PMID- 16251987 TI - Genetic influences on impulsivity, risk taking, stress responsivity and vulnerability to drug abuse and addiction. AB - Genetic variation may partially underlie complex personality and physiological traits--such as impulsivity, risk taking and stress responsivity--as well as a substantial proportion of vulnerability to addictive diseases. Furthermore, personality and physiological traits themselves may differentially affect the various stages of addiction, defined chronologically as initiation of drug use, regular drug use, addiction/dependence and potentially relapse. Here we focus on recent approaches to the study of genetic variation in these personality and physiological traits, and their influence on and interaction with addictive diseases. PMID- 16251988 TI - Decision making, impulse control and loss of willpower to resist drugs: a neurocognitive perspective. AB - Here I argue that addicted people become unable to make drug-use choices on the basis of long-term outcome, and I propose a neural framework that explains this myopia for future consequences. I suggest that addiction is the product of an imbalance between two separate, but interacting, neural systems that control decision making: an impulsive, amygdala system for signaling pain or pleasure of immediate prospects, and a reflective, prefrontal cortex system for signaling pain or pleasure of future prospects. After an individual learns social rules, the reflective system controls the impulsive system via several mechanisms. However, this control is not absolute; hyperactivity within the impulsive system can override the reflective system. I propose that drugs can trigger bottom-up, involuntary signals originating from the amygdala that modulate, bias or even hijack the goal-driven cognitive resources that are needed for the normal operation of the reflective system and for exercising the willpower to resist drugs. PMID- 16251989 TI - Nicotine addiction and comorbidity with alcohol abuse and mental illness. AB - The World Health Organization estimates that one-third of the global adult population smokes. Because tobacco use is on the rise in developing countries, death resulting from tobacco use continues to rise. Nicotine, the main addictive component of tobacco, initiates synaptic and cellular changes that underlie the motivational and behavioral alterations that culminate in addiction. Nicotine addiction progresses rapidly in adolescents and is most highly expressed in vulnerable people who have psychiatric illness or other substance abuse problems. PMID- 16251990 TI - Laboratory models of alcoholism: treatment target identification and insight into mechanisms. AB - Laboratory models, including animal tissues and live animals, have proven useful for discovery of molecular targets of alcohol action as well as for characterization of genetic and environmental factors that influence alcohol's neural actions. Here we consider strengths and weaknesses of laboratory models used in alcohol research and analyze the limitations of using animals to model a complex human disease. We describe targets for the neural actions of alcohol, and we review studies in which animal models were used to examine excessive alcohol drinking and to discover genes that may contribute to risk for alcoholism. Despite some limitations of the laboratory models used in alcohol research, these experimental approaches are likely to contribute to the development of new therapies for alcohol abuse and alcoholism. PMID- 16251991 TI - Neural systems of reinforcement for drug addiction: from actions to habits to compulsion. AB - Drug addiction is increasingly viewed as the endpoint of a series of transitions from initial drug use--when a drug is voluntarily taken because it has reinforcing, often hedonic, effects--through loss of control over this behavior, such that it becomes habitual and ultimately compulsive. Here we discuss evidence that these transitions depend on interactions between pavlovian and instrumental learning processes. We hypothesize that the change from voluntary drug use to more habitual and compulsive drug use represents a transition at the neural level from prefrontal cortical to striatal control over drug seeking and drug taking behavior as well as a progression from ventral to more dorsal domains of the striatum, involving its dopaminergic innervation. These neural transitions may themselves depend on the neuroplasticity in both cortical and striatal structures that is induced by chronic self-administration of drugs. PMID- 16251992 TI - Effect of ginseng saponins on enhanced dopaminergic transmission and locomotor hyperactivity induced by nicotine. AB - Several studies have shown that behavioral hyperactivity induced by psychomotor stimulants is prevented by ginseng saponins. In an attempt to investigate whether the effect of ginseng saponins is through their inhibitory action on the enhanced dopaminergic transmission by psychomotor stimulants, we examined the effects of ginseng total saponin (GTS) presynaptically on nicotine-induced dopamine (DA) release in the striatum of freely moving rats using in vivo microdialysis technique and postsynaptically on the in vitro and in vivo binding of [3H]raclopride to DA D2 receptors. Also, we examined the effects of GTS on nicotine-induced locomotor hyperactivity and on nicotine-induced Fos protein expression in the nucleus accumbens and striatum. Systemic pretreatment with GTS (100 and 400 mg/kg, intraperitoneally (i.p.)) resulted in a dose-dependent inhibition of locomotor hyperactivity induced by nicotine. GTS decreased nicotine induced DA release in the striatum in a dose-dependent manner. However, GTS had no effects on resting levels of locomotor activity and extracellular DA in the striatum. GTS inhibited the in vitro binding of [3H]raclopride to rat striatal membranes with an IC50 of 5.14+/-1.09 microM. High doses of GTS (400 and 800 mg/kg, i.p.) resulted in decreases in the in vivo binding of [3H]raclopride in the striatum. GTS decreased nicotine-induced Fos protein expression in the nucleus accumbens and striatum, reflecting the inhibition by GTS of nicotine induced enhancement of dopaminergic transmission. The results of the present study suggest that GTS acts not only on dopaminergic neurons directly or indirectly to prevent nicotine-induced DA release but also postsynaptically by binding to DA D2 receptors. This may explain the blocking effect of GTS on behavioral activation induced by nicotine and conceivably by other psychostimulants. Our data raise the possibility that GTS, by attenuating nicotine-induced enhancement of dopaminergic transmission, may prove to be a useful therapeutic agent for nicotine addiction and warrant further investigation on its effect on nicotine's rewarding property. PMID- 16251993 TI - Neuropsychological profile of acute alcohol intoxication during ascending and descending blood alcohol concentrations. AB - Numerous studies have investigated the effects of alcohol on motor processes during rising and declining blood alcohol concentrations (BAC), however, relatively little research has examined the alcohol-induced impairment of cognitive performance on the two limbs of the BAC curve. This experiment administered a neuropsychological test battery to assess the degree to which rising and declining BACs during an acute dose of alcohol impair nine cognitive processes within an individual. In all, 20 healthy male social drinkers (university students) were assigned to one of two groups (n = 10) who received a beverage containing either 0.0 g/kg (placebo) or 0.65 g/kg alcohol and performed the test battery when BAC was increasing and was decreasing. Comparisons of alcohol and placebo groups revealed impairment (slower response and/or increased errors) in seven of the cognitive processes: long-term verbal memory; information processing; declarative memory; inhibitory control; short-term visual memory; long-term visual memory, and visual-spatial working memory. However, some processes were impaired only during rising BACs whereas the impairment of others during declining BACs was evident only by an increase in errors. These results show cognitive tasks performed by an individual are not similarly affected by rising and declining BACs, and call attention to the importance of assessing both speed and accuracy on both limbs of the BAC curve. The particular cognitive processes differentially affected by rising vs declining BACs raised the possibility that acute alcohol intoxication may impair one cerebral hemisphere to a greater degree than the other, and this could be explored by neuroimaging techniques. PMID- 16251994 TI - Dominant-negative c-Jun inhibits rat cardiac hypertrophy induced by angiotensin II and hypertension. AB - Cardiac activator protein-1 (AP-1), composed of c-Jun, is significantly activated by hypertension or angiotensin II (AngII). This study was undertaken to elucidate whether c-Jun could be the potential target for treatment of cardiac hypertrophy. We constructed recombinant adenovirus carrying dominant-negative mutant of c-Jun (Ad.DN-c-Jun). Using catheter-based technique of adenoviral gene transfer, we achieved global myocardial transduction of DN-c-Jun in rats, to specifically inhibit cardiac AP-1. (1) AngII (200 ng/kg/min) infusion in rats caused cardiac hypertrophy, increased cardiac p70S6 kinase activity by 1.3-fold (P<0.05) and enhanced the gene expression of cardiac hypertrophic markers. Ad.DN-c-Jun, which was transferred to the heart 2 days before AngII infusion, prevented cardiac hypertrophy (P<0.01), decreased p70S6 kinase phosphorylation (P<0.05), and suppressed cardiac gene expression of brain natriuretic peptide, collagen I, III, and IV, monocyte chemoattractant protein-1 (MCP-1) and plasminogen activator inhibitor-1 (PAI-1) (P<0.01). (2) In genetically hypertensive rats with cardiac hypertrophy, cardiac gene transfer of Ad.DN-c-Jun, without affecting hypertension, regressed cardiac hypertrophy (P<0.05), and suppressed p70S6 kinase phosphorylation by 20% (P<0.05) and suppressed the enhanced expression of collagen I, III, and IV, MCP-1 and PAI-1. These results provided the first evidence that in vivo blockade of cardiac c-Jun inhibits pathologic cardiac hypertrophy. PMID- 16251995 TI - Prevention of posterior capsule opacification by the induction of therapeutic apoptosis of residual lens cells. AB - Posterior capsule opacification (PCO) is a common complication of cataract surgery. Using adenovirus(Ad)-mediated gene transfer, we overexpressed the proapoptotic molecules p53, procaspase 3, Bax, and TRAIL to induce therapeutic programmed cell death of residual lens cells to prevent PCO. Overexpressed TRAIL did not induce apoptosis in cultured rabbit lens cells or in human lens cells. Overexpressed p53 induced apoptosis of lens cells in vitro and ex vivo, but was unable to prevent PCO in vivo. Overexpressed procaspase 3 was associated with engagement of many components of the apoptotic pathway, including cleavage of intracellular caspase targets such as PARP and inter-nucleosome DNA fragmentation. Even when only slightly overexpressed, Bax caused apoptosis of transduced rabbit and human lens cells by engaging the mitochondrial pathway, including catalytic activation of the caspases. A single in vivo injection of Ad vectors expressing either Bax or procaspase 3 into the capsular bag at the end of phacoemulsification prevented PCO in rabbits. These experiments show that Ad mediated Bax or procaspase 3 overexpression is capable of inducing therapeutic programmed cell death in vitro and in vivo in residual lens cells and preventing PCO in a rabbit model of PCO. Manipulation of proapoptotic molecule expression could be a novel gene therapy approach for prevention of PCO. PMID- 16251996 TI - Adenoviral GDNF gene transfer enhances neurofunctional recovery after recurrent laryngeal nerve injury. AB - To assess the possibility of gene therapy for recurrent laryngeal nerve (RLN) injury, we examined functional and histological recovery after glial cell line derived neurotrophic factor (GDNF) gene transfer in a rat RLN crush model. Adenoviral vector encoding beta-galactosidase gene (AxCALacZ) or human GDNF gene (AxCAhGDNF) was injected into the crush site of the RLN. Neurons in the nucleus ambiguus on the crushed side were labeled with X-gal or GDNF immnohistochemistry after AxCALacZ or AxCAhGDNF injection. Reverse transcription-polymerase chain reaction analysis revealed expression of human GDNF mRNA transcripts in brainstem tissue containing the nucleus ambiguus on the crushed side after AxCAhGDNF injection. Animals injected with AxCAhGDNF displayed significantly improved motor nerve conduction velocity of the RLN and recovery rate of vocal fold movement at 2 and 4 weeks after treatment as compared to controls. AxCAhGDNF-injected animals showed a significantly larger axonal diameter and improved remyelination in crushed RLN as compared to controls. Adenoviral GDNF gene transfer may thus promote laryngeal function recovery after RLN injury. Inoculation of adenoviral vector containing the GDNF gene at the site of damage soon after nerve injury may assist patients with laryngeal paralysis caused by nerve injury during head and neck surgery. PMID- 16251998 TI - Dose-dependent improvements in outcome with adenoviral expression of interleukin 10 in a murine model of multisystem organ failure. AB - Targeted expression of interleukin-10 (IL-10) has been proposed as a means to suppress acute and chronic inflammation. We explored the capacity of targeted adenoviral expression of human or viral IL-10 to improve outcome in a zymosan induced model of acute lung injury and multisystem organ failure. Intratracheal administration of adenovirus expressing either human or viral IL-10 prior to zymosan administration significantly improved survival at a dose of 10(7) particles (P<0.01), whereas the same recombinant vectors were ineffective at 10(8) particles and increased mortality at 10(9) particles. Improved survival after administration of 10(7) particles of adenovirus expressing viral or human IL-10 was associated with local tissue expression of IL-10 (100-300 pg/g wet wt). In contrast, mortality after administration of 10(9) particles was associated with markedly elevated IL-10 expression, both in the lung (10000-70000 pg/g wet wt) and systemically (1000-3000 pg/ml plasma), with evidence of an exaggerated systemic inflammatory response (plasma IL-6 and TNFalpha). Targeted gene expression of IL-10 can be used to treat acute inflammatory processes, but increased doses resulting in its systemic release are not associated with improvements in outcome, and may actually exacerbate acute inflammatory processes. PMID- 16251999 TI - The epidemiology of blood pressure in East Asia. PMID- 16251997 TI - p27(SJ), a novel protein in St John's Wort, that suppresses expression of HIV-1 genome. AB - Transcription of the HIV-1 genome is controlled by the cooperation of viral regulatory proteins and several host factors which bind to specific DNA sequences within the viral promoter spanning the long terminal repeat, (LTR). Here, we describe the identification of a novel protein, p27(SJ), present in a laboratory callus culture of Hypericum perforatum (St John's Wort) that suppresses transcription of the HIV-1 genome in several human cell types including primary culture of microglia and astrocytes. p27(SJ) associates with C/EBPbeta, a transcription factor that regulates expression of the HIV-1 genome in macrophages and monocytic cells, and the viral transactivator, Tat. The association of p27(SJ) with C/EBPbeta and Tat alters their subcellular localization, causing their accumulation in the perinuclear cytoplasmic compartment of the cells. Fusion of a nuclear localization signal to p27(SJ) forces its entry into the nucleus and diminishes the capacity of p27(SJ) to suppress Tat activity, but does not alter its ability to suppress C/EBPbeta activation of the LTR. Results from binding assays showed the inhibitory effect of p27(SJ) on C/EBPbeta interaction with DNA. Finally, our results demonstrate that expression of p27(SJ) decreases the level of viral replication in HIV-1-infected cells. These observations suggest the potential for the development of a therapeutic advance based on p27(SJ) protein to control HIV-1 transcription and replication in cells associated with HIV-1 infection in the brain. PMID- 16252000 TI - Haplotype association of the adrenergic beta2 receptor gene with essential hypertension. PMID- 16252001 TI - Aim, design and methods of the 'reasons for not intensifying antihypertensive treatment' (RIAT): an international registry in essential hypertension. AB - Failure of physicians to adhere to hypertension guidelines may partly account for the failure to achieve blood pressure (BP) goals in clinical practice. The aim of this trial is a comprehensive description of the approach of physicians in the management of high BP among primary care patients. It will primarily assess what are the Reasons for not Intensifying an Antihypertensive Treatment (RIAT), when predefined individual BP goals are not achieved. Open intervention survey was conducted in 17 countries in Latin America, Eastern Europe, Africa and Asia in family practices, government and private clinics. The registry is based on a three-step epidemiological design. Step one shall identify guidelines and recommendations taken as reference in each country for the management of hypertension. Step two will assess the variance between individual targets defined by physicians in their practice compared to guidelines and recommendations. Step three is a prospective registry where physicians collect patient data at baseline; determine individual target BP values. Several follow up visits are proposed to monitor achievement of these targets. Step three of RIAT aims at providing responses to several key objectives. Recruitment is under way aiming at enrolling 33,000 patients. To identify, what is the BP targeted according to the risk factor profile and what are the reasons for not modifying an antihypertensive treatment when BP goals are not reached, and to analyse the type of antihypertensive drugs prescribed according to compelling indications and to assess the percentage of patients reaching target figures. PMID- 16252002 TI - The amyloid-beta precursor protein: integrating structure with biological function. AB - Proteolytic processing of the amyloid-beta precursor protein (APP) generates the Abeta amyloid peptide of Alzheimer's disease. The biological function of APP itself remains, however, unclear. In the current review, we study in detail the different subdomains of APP and try to assign functional significance to particular structures identified in the protein. PMID- 16252003 TI - Structure of the split PH domain and distinct lipid-binding properties of the PH PDZ supramodule of alpha-syntrophin. AB - Pleckstrin homology (PH) domains play diverse roles in cytoskeletal dynamics and signal transduction. Split PH domains represent a unique subclass of PH domains that have been implicated in interactions with complementary partial PH domains 'hidden' in many proteins. Whether partial PH domains exist as independent structural units alone and whether two halves of a split PH domain can fold together to form an intact PH domain are not known. Here, we solved the structure of the PH(N)-PDZ-PH(C) tandem of alpha-syntrophin. The split PH domain of alpha syntrophin adopts a canonical PH domain fold. The isolated partial PH domains of alpha-syntrophin, although completely unfolded, remain soluble in solution. Mixing of the two isolated domains induces de novo folding and yields a stable PH domain. Our results demonstrate that two complementary partial PH domains are capable of binding to each other to form an intact PH domain. We further showed that the PH(N)-PDZ-PH(C) tandem forms a functionally distinct supramodule, in which the split PH domain and the PDZ domain function synergistically in binding to inositol phospholipids. PMID- 16252004 TI - Critical role of novel Thr-219 autophosphorylation for the cellular function of PKCtheta in T lymphocytes. AB - Phosphopeptide mapping identified a major autophosphorylation site, phospho (p)Thr-219, between the tandem C1 domains of the regulatory fragment in protein kinase C (PKC)theta. Confirmation of this identification was derived using (p)Thr 219 antisera that reacted with endogenous PKCtheta in primary CD3+ T cells after stimulation with phorbol ester, anti-CD3 or vanadate. The T219A mutation abrogated the capacity of PKCtheta to mediate NF-kappaB, NF-AT and interleukin-2 promoter transactivation, and reduced PKCtheta's ability in Jurkat T cells to phosphorylate endogenous cellular substrates. In particular, the T219A mutation impaired crosstalk of PKCtheta with Akt/PKBalpha in NF-kappaB activation. Yet, this novel (p)Thr-219 site did not affect catalytic activity or second-messenger lipid-binding activity in vitro. Instead, the T219A mutation prevented proper recruitment of PKCtheta in activated T cells. The PKCthetaT219A mutant defects were largely rescued by addition of a myristoylation signal to force its proper membrane localization. We conclude that autophosphorylation of PKCtheta at Thr 219 plays an important role in the correct targeting and cellular function of PKCtheta upon antigen receptor ligation. PMID- 16252005 TI - Transactivation of Schizosaccharomyces pombe cdt2+ stimulates a Pcu4-Ddb1-CSN ubiquitin ligase. AB - Cullin-4 forms a scaffold for multiple ubiquitin ligases. In Schizosaccharomyces pombe, the Cullin-4 homologue (Pcu4) physically associates with Ddb1 and the COP9 signalosome (CSN). One target of this complex is Spd1. Spd1 regulates ribonucleotide reductase (RNR) activity. Spd1 degradation during S phase, or following DNA damage of G2 cells, results in the nuclear export of the small RNR subunit. We demonstrate that Cdt2, an unstable WD40 protein, is a regulatory subunit of Pcu4-Ddb1-CSN ubiquitin ligase. cdt2 deletion stabilises Spd1 and prevents relocalisation of the small RNR subunit from the nucleus to the cytoplasm. cdt2+ is periodically transcribed by the Cdc10/DSC1 transcription factor during S phase and transiently transcribed following DNA damage of G2 cells, corresponding to Spd1 degradation profiles. Cdt2 co-precipitates with Spd1, and Cdt2 overexpression results in constitutive Spd1 degradation. We propose that Cdt2 incorporation into the Pcu4-Ddb1-CSN complex prompts Spd1 targeting and subsequent degradation and that Cdt2 is a WD40 repeat adaptor protein for Cullin-4-based ubiquitin ligase. PMID- 16252007 TI - Linking the group II intron catalytic domains: tertiary contacts and structural features of domain 3. AB - Despite its importance for group II intron catalytic activity, structural information on conserved domain 3 (D3) is extremely limited. This domain is known to specifically stimulate the chemical rate of catalysis and to function as a 'catalytic effector'. Of all the long-range tertiary contacts that have been identified within group II introns, none has included D3 residues. Furthermore, little is known about the atoms and functional groups in D3 that contribute to catalysis. Using a nucleotide analog interference mapping assay with an extended repertoire of nucleotide analogs, we have identified functional groups in D3 that are critical for ribozyme activity. These data, together with mutational analysis, suggest the formation of noncanonical base pairs within the phylogenetically conserved internal loop at the base of D3. Finally, a related nucleotide analog interference suppression study resulted in the identification of a direct tertiary interaction between D3 and catalytic domain 5, which sheds new light on D3 function in the group II intron structure and mechanism. PMID- 16252006 TI - Ligand-induced transrepression by VDR through association of WSTF with acetylated histones. AB - We have previously shown that the novel ATP-dependent chromatin-remodeling complex WINAC is required for the ligand-bound vitamin D receptor (VDR)-mediated transrepression of the 25(OH)D3 1alpha-hydroxylase (1alpha(OH)ase) gene. However, the molecular basis for VDR promoter association, which does not involve its binding to specific DNA sequences, remains unclear. To address this issue, we investigated the function of WSTF in terms of the association between WINAC and chromatin for ligand-induced transrepression by VDR. Results of in vitro experiments using chromatin templates showed that the association of unliganded VDR with the promoter required physical interactions between WSTF and both VDR and acetylated histones prior to VDR association with chromatin. The acetylated histone-interacting region of WSTF was mapped to the bromodomain, and a WSTF mutant lacking the bromodomain served as a dominant-negative mutant in terms of ligand-induced transrepression of the 1alpha(OH)ase gene. Thus, our findings indicate that WINAC associates with chromatin through a physical interaction between the WSTF bromodomain and acetylated his tones, which appears to be indispensable for VDR/promoter association for ligand-induced transrepression of 1alpha(OH)ase gene expression. PMID- 16252008 TI - Transcriptome analysis reveals cyclobutane pyrimidine dimers as a major source of UV-induced DNA breaks. AB - Photolyase transgenic mice have opened new avenues to improve our understanding of the cytotoxic effects of ultraviolet (UV) light on skin by providing a means to selectively remove either cyclobutane pyrimidine dimers (CPDs) or pyrimidine (6-4) pyrimidone photoproducts. Here, we have taken a genomics approach to delineate pathways through which CPDs might contribute to the harmful effects of UV exposure. We show that CPDs, rather than other DNA lesions or damaged macromolecules, comprise the principal mediator of the cellular transcriptional response to UV. The most prominent pathway induced by CPDs is that associated with DNA double-strand break (DSB) signalling and repair. Moreover, we show that CPDs provoke accumulation of gamma-H2AX, P53bp1 and Rad51 foci as well as an increase in the amount of DSBs, which coincides with accumulation of cells in S phase. Thus, conversion of unrepaired CPD lesions into DNA breaks during DNA replication may comprise one of the principal instigators of UV-mediated cytotoxicity. PMID- 16252009 TI - Unstable microtubule capture at kinetochores depleted of the centromere associated protein CENP-F. AB - Centromere protein F (CENP-F) (or mitosin) accumulates to become an abundant nuclear protein in G2, assembles at kinetochores in late G2, remains kinetochore bound until anaphase, and is degraded at the end of mitosis. Here we show that the absence of nuclear CENP-F does not affect cell cycle progression in S and G2. In a subset of CENP-F depleted cells, kinetochore assembly fails completely, thereby provoking massive chromosome mis-segregation. In contrast, the majority of CENP-F depleted cells exhibit a strong mitotic delay with reduced tension between kinetochores of aligned, bi-oriented sister chromatids and decreased stability of kinetochore microtubules. These latter kinetochores generate mitotic checkpoint signaling when unattached, recruiting maximum levels of Mad2. Use of YFP-marked Mad1 reveals that throughout the mitotic delay some aligned, CENP-F depleted kinetochores continuously recruit Mad1. Others rebind YFP-Mad1 intermittently so as to produce 'twinkling', demonstrating cycles of mitotic checkpoint reactivation and silencing and a crucial role for CENP-F in efficient assembly of a stable microtubule-kinetochore interface. PMID- 16252011 TI - What standards apply to resuscitation at the borderline of gestational age? PMID- 16252012 TI - Physicians' refusal to resuscitate at borderline gestational age. AB - Most neonatologists believe there is a minimal gestational age, below which it is appropriate to refuse to provide resuscitation or intensive care. Determination of this threshold should involve knowledge of the outcome data, but also an understanding of the potential for misuse of these data. In particular, there is a risk of deception, of the parents and of ourselves, due to the uncertainty of the true gestational age, and the "self-fulfilling prophecy" that may occur when a center refuses to try below a certain gestational age because they have had no survivors below that age. Finally, any refusal to treat requires ethical justification. Concepts such as futility and patient's best interest should play a role in the determination of the gestational age threshold, applied in light of the data's inherent weaknesses. PMID- 16252013 TI - Perinatal/neonatal case presentation: unexpected severe respiratory insufficiency in a newborn with Holt-Oram Syndrome. AB - Holt-Oram syndrome is an autosomal dominant condition characterized by skeletal and cardiac defects. Pulmonary malformation is not reported to belong to the spectrum of this condition. We report a second case of a newborn with Holt-Oram syndrome who developed severe respiratory insufficiency shortly after birth. We discuss possible genetic links between abnormal pulmonary morphogenesis and Holt Oram syndrome. PMID- 16252010 TI - TRAF6-mediated ubiquitination regulates nuclear translocation of NRIF, the p75 receptor interactor. AB - TRAF6 is an E3 ubiquitin ligase that mediates signaling from members of the tumor necrosis factor and Toll-like receptor superfamilies, including the p75 neurotrophin receptor. Recently, TRAF6 was shown to bind to another p75 cytoplasmic interactor, NRIF, and promote its nuclear localization. Here, we demonstrate that NRIF is a substrate for TRAF6-mediated K63 polyubiquitination and that this modification is necessary for its nuclear translocation. Activation of p75 resulted in NRIF polyubiquitination, association with TRAF6 and nuclear localization. NRIF was polyubiquitinated by TRAF6 in vitro and in cultured cells, and this was abrogated by mutation of K19 in the amino-terminus of NRIF. The K19R mutant NRIF displayed reduced TRAF6 association and neurotrophin-dependent nuclear localization. In neurons from traf6-/- mice, NRIF failed to enter the nucleus in response to p75 activation, and polyubiquitination and nuclear localization were attenuated in traf6-/- brain. Finally, unlike wild-type NRIF, the K19R NRIF failed to reconstitute p75-mediated apoptosis in nrif-/- neurons. These results reveal a unique mechanism of p75 signaling and a novel role for K63 linked ubiquitin chains. PMID- 16252014 TI - Disodium disuccinate astaxanthin (Cardax): antioxidant and antiinflammatory cardioprotection. AB - Disodium disuccinate astaxanthin (Cardax), DDA) has cardioprotective effects in the rat, rabbit, and canine models of experimental infarction. It is highly effective by parenteral administration in subchronic and acute dosing regimens. Unpublished data in rats suggest that oral cardioprotection is also readily achievable. DDA-induced myocardial salvage in the canine can reach 100% with a 4 day subchronic dosing regimen. At a single i.v. dose DDA is cardioprotective, when given 2 h before experimental coronary occlusion, but the protection is on the average two-thirds of that achieved with the subchronic regimen in dogs. In conscious animals DDA has no effects on hemodynamic parameters. The primary mechanism of cardioprotection appears to be antioxidant activity involving direct scavenging of superoxide anion, the lynchpin radical in ischemia-reperfusion injury. In addition, modulation of serum complement activity, as well as the reduction in the levels of C-reactive protein (CRP) and the membrane attack complex (MAC) in infarcted tissue suggest a significant antiinflammatory component in the mechanism of cardioprotective action of DDA. Stoichiometric binding of the meso-form of the compound to human serum albumin (HSA) has been demonstrated in vitro. This binding capacity overcomes the supramolecular assembly of the compound in aqueous solution, which by itself improves the stability and shelf life of aqueous formulations. Non-esterified astaxanthin readily enters cardiac tissue after either oral or parenteral administration, providing a reservoir of a cardioprotective agent with a significant half-life due to favorable ADME in mammals. Due to the well-documented safety profile of non-esterified astaxanthin in humans, disodium disuccinate astaxanthin may well find clinical utility in cardiovascular indications in humans following successful completion of preclinical and clinical pharmacology and toxicology studies. PMID- 16252015 TI - The novel antiarrhythmic drug dronedarone: comparison with amiodarone. AB - Dronedarone is a noniodinated benzofuran derivative that has been developed to overcome the limiting iodine-associated adverse effects of the commonly used antiarrhythmic drug, amiodarone. It displays a wide cellular electrophysiological spectrum largely similar to amiodarone, inhibiting the potassium currents I(Kr), I(Ks), I(KI), I(KACh), and I(sus), as well as sodium currents and L-type calcium currents in isolated cardiomyocytes. In addition, dronedarone exhibits antiadrenergic properties. In vivo, dronedarone has been shown to be more effective than amiodarone in several arrhythmia models, particularly in preventing ischemia- and reperfusion-induced ventricular fibrillation and in reducing mortality. However, an increased incidence of torsades de pointes with dronedarone in dogs shows that possible proarrhythmic effects of dronedarone require further evaluation. The clinical trails DAFNE, EURIDIS, and ADONIS indicated safety, antiarrhythmic efficacy and low proarrhythmic potential of the drug in low-risk patients. In contrast, the increased incidence of death in the dronedarone group of the discontinued ANDROMEDA trial raises safety concerns for patients with congestive heart failure and moderate to severe left ventricular dysfunction. Dronedarone appears to be effective in preventing relapses of atrial fibrillation and atrial flutter. Torsades de pointes, the most severe adverse effect associated with amiodarone, has not yet been reported in humans with dronedarone. Unlike amiodarone, dronedarone had little effect on thyroid function and hormone levels in animal models and had no significant effects on human thyroid function in clinical trials. In conclusion, dronedarone could be a useful drug for prevention of atrial fibrillation and atrial flutter relapses in low risk patients. However, further experimental studies and long-term clinical trials are required to provide additional evidence of efficacy and safety of dronedarone. PMID- 16252016 TI - Bimatoprost: mechanism of ocular surface hyperemia associated with topical therapy. AB - Bimatoprost is a safe and well-tolerated intraocular pressure (IOP) lowering drug that was approved in the United States in 2001 for the treatment of glaucoma and ocular hypertension. It is highly efficacious and produces greater mean reductions in IOP than other currently available antiglaucoma drugs. Conjunctival hyperemia is a common side effect of bimatoprost, but the hyperemia is typically mild and transient. No association has been found between signs of inflammation and the presence of hyperemia in bimatoprost-treated patients. Preclinical studies have elucidated the pharmacological mechanism of bimatoprost-related hyperemia and have examined the possible involvement of inflammation. Bimatoprost, as well as the free acid of latanoprost, elicited endothelium dependent vasorelaxation in the rabbit jugular vein preparation, a quantitative in vitro model for ocular surface hyperemia (OSH). The vasorelaxant responses to either bimatoprost or latanoprost free acid were significantly inhibited by L NAME, a nitric oxide synthase inhibitor. Similarly, the in vivo OSH responses to topically applied bimatoprost or latanoprost in dog eyes were significantly inhibited by L-NAME. As predicted, prostaglandin E(2) (PGE(2))-induced conjunctival hyperemia was not inhibited by L-NAME, since PGE(2) has a direct relaxant effect on the vascular smooth muscle. In-life observations and histopathological assessment of ocular surface tissues following bimatoprost treatment were performed for multiple-dose one month, 6 month, or 12 month safety studies in rabbits, dogs, and non-human primates. Results of these studies showed no evidence of bimatoprost-related inflammation in the ocular surface tissues. In summary, OSH related to bimatoprost treatment in laboratory animals occurs by endothelial-derived nitric oxide-mediated vasodilatation and is not associated with inflammation. These studies suggest that conjunctival hyperemia, a side effect of bimatoprost treatment, results from non-inflammatory, pharmacologically based vasodilatation. PMID- 16252017 TI - Role of mitogen-activated protein (MAP) kinases in cardiovascular diseases. AB - Over the last decade important advances have been made in our understanding of the molecular events underlying cellular responses to extracellular signals. Increased understanding of signal transduction mechanisms and gene regulation involved in cardiovascular diseases has created opportunities for the discovery of novel therapeutic compounds useful for the treatment of cardiovascular disorders. One of the best-studied signalling routes is the mitogen activated protein (MAP) kinase signal transduction pathway, which plays a crucial role in many aspects of cardiovascular responses. Here, our current understanding of the MAP kinase pathway is reviewed, as well as recent advances in the design of novel agents that are able to modulate the activity of these signaling cascades. PMID- 16252018 TI - Go 6983: a fast acting protein kinase C inhibitor that attenuates myocardial ischemia/reperfusion injury. AB - Reperfusion injury is characterized by a decrease in endothelial release of nitric oxide within 5 min after reperfusion, increased leukocyte-endothelium interaction, and transmigration of leukocytes into the myocardium, producing cardiac contractile dysfunction. Go 6983 is a fast acting, lipid soluble, broad spectrum protein kinase C inhibitor. When administered at the beginning of reperfusion, it can restore cardiac function within 5 min and attenuate the deleterious effects associated with acute ischemia/reperfusion. Go 6983 may offer greater cardioprotection than other broad-spectrum PKC inhibitors in postischemic reperfusion injury because it inhibits PKC(zeta) as well as four other isoforms. The cardioprotection is associated with decreased leukocyte superoxide release and increased endothelial derived nitric oxide from vascular tissue. In vitro studies of human tissue showed that Go 6983 significantly inhibited antigen induced superoxide release from leukocytes of patients previously sensitized to tree pollen. In human vascular tissue, Go 6983 inhibited intracellular Ca(2+) accumulation, suggesting a mechanism for its vasodilator properties. These studies suggest that Go 6983 would be an effective compound to use in a clinical ischemia/reperfusion setting of organ transplantation and/or cerebral ischemia where inhibiting superoxide release and vasoconstriction in postischemic tissues would allow for better restoration of organ function during reperfusion. However, given the broad-spectrum action of Go 6983, careful titration of the dose regimen would be recommended to ensure a successful outcome in the setting of organ transplantation and/or cerebral ischemia. PMID- 16252019 TI - From Quinlan to Schiavo: medical, ethical, and legal issues in severe brain injury. PMID- 16252020 TI - New frontiers in radiosurgery for the brain and body. AB - Radiosurgery is defined as the use of highly focused beams of radiation to ablate a pathologic target, thus achieving a surgical objective by noninvasive means. Recent advances have allowed a wide variety of intracranial lesions to be effectively treated with radiosurgery, and radiosurgical treatment has been accepted as a standard part of the neurosurgical armamentarium. The advent of frameless radiosurgery now permits radiosurgical treatment to all parts of the body and is being actively explored by many centers. This article reviews some of the modern tools for radiosurgical treatment and discusses the current clinical practice of radiosurgery. PMID- 16252023 TI - Use of PET/CT scanning in cancer patients: technical and practical considerations. AB - This overview of the oncologic applications of positron emission tomography (PET) focuses on the technical aspects and clinical applications of a newer technique: the combination of a PET scanner and a computed tomography (CT) scanner in a single (PET/CT) device. Examples illustrate how PET/CT contributes to patient care and improves upon the previous state-of-the-art method of comparing a PET scan with a separate CT scan. Finally, the author presents some of the results from studies of PET/CT imaging that are beginning to appear in the literature. PMID- 16252024 TI - The variability of platelet response to aspirin and clopidogrel: revisiting the Caprie, Cure, Credo, and Match trials. AB - In a significant number of patients, platelets do not respond to aspirin and/or clopidogrel. Furthermore, this lack of response has recently been shown to affect cardiovascular outcome. At the time of the CAPRIE, CURE, CREDO, and MATCH studies, no in vitro assessment was made to determine platelet response. In vitro platelet response should be considered as an important correctable risk factor for atherosclerotic events. PMID- 16252025 TI - Renal cell carcinoma: diagnosis and treatment, 1994-2003. PMID- 16252026 TI - The troubles with pinkies. PMID- 16252028 TI - Thomas Hodgkin: medical immortal and uncompromising idealist. PMID- 16252029 TI - Osler's role in defining the third corpuscle, or "blood plates". AB - As a young microscopist in 1874, William Osler was probably the first physician to recognize platelets in blood as a single unit and their coalescence when blood was shed. He expounded on the earlier observations of Schultze, who in 1865 noted abundant, irregular masses of colorless globules in normal blood that were almost certainly platelets. The impact of Osler's initial observations were diminished by an unclear relation between this third blood particle and bacteria. Initial observers of this blood element were handicapped by the inadequacy of microscopes, the lack of anticoagulants and blood stains, and the tendency of platelets to clump and undergo morphologic change when blood is shed. PMID- 16252030 TI - Robert Lee Fine, MD: a conversation with the editor. Interview by William Clifford Roberts. PMID- 16252027 TI - Abdominal aortic aneurysm (AAA): cost-effectiveness of screening, surveillance of intermediate-sized AAA, and management of symptomatic AAA. AB - Physicians must make decisions about screening patients for abdominal aortic aneurysms (AAAs), monitoring or referring for surgery patients with AAAs of various sizes, and assessing patients with symptoms that may be related to AAAs. This review article analyzes the evidence for each scenario. The effectiveness and cost-effectiveness of screening for AAA is based on results from four randomized controlled trials. A cost-effectiveness analysis using a Markov model showed that ultrasound screening of white men beginning at age 65 is both effective and cost-effective in preventing AAA-related death. Such screening would have a small but real impact over a 20-year period in these men. For patients with a known AAA-which is often detected incidentally-the evidence clearly suggests periodic ultrasound surveillance for those with small AAAs (3.0 3.9 cm in diameter) and elective surgical repair for those with large AAAs (>or=5.5 cm). Two recent randomized controlled trials have shown that early surgical repair confers no survival benefit compared with periodic surveillance for patients with intermediate-sized AAAs (4.0-5.5 cm in diameter), so those patients can also be monitored. Some centers choose to increase the frequency of monitoring to every 3 to 6 months when the AAA reaches 5.0 cm. Factors to consider in assessing symptomatic patients include the high risk of life threatening conditions, the potential increased risk of death or poor outcome with delay in diagnosis, the limitations of ultrasound in identifying whether symptoms are due to known or suspected AAA, and the timely availability of computed tomography or other imaging tests. If available, computed tomography is preferred in patients with recent or severe symptoms, since it is better at detecting retroperitoneal hemorrhage and other complications and in providing preoperative definition of the anatomy. PMID- 16252031 TI - Development and implementation of a pharmacist-managed inpatient warfarin protocol. AB - Because of the complexities of warfarin administration and its bleeding complications, a pharmacist-managed protocol for warfarin administration was developed at Baylor University Medical Center. The protocol incorporated current clinical guidelines and evidence-based medicine. Clinical outcomes under the protocol were compared with those of usual care, physician management, in a pilot study. Twenty-nine patients were enrolled in the protocol group and 22 in the control group between August 2004 and November 2004. Results showed that patients in both groups achieved therapeutic ranges of warfarin within 6 days. However, the pharmacist-managed patients exhibited a trend toward fewer adverse drug reactions (7% vs 14%) and fewer supratherapeutic international normalized ratios (17% vs 27%) than the control group, although the difference was not statistically significant. Based on these results and the results of similar larger studies showing the effectiveness of pharmacist-managed warfarin administration, Baylor University Medical Center made the protocol available for use in the hospital in May 2005. PMID- 16252032 TI - Painful nodule with induration and spreading erythema. PMID- 16252033 TI - Abnormal electrocardiogram obtained after an automobile accident. PMID- 16252034 TI - Critical left main coronary artery stenosis diagnosed by computed tomographic coronary angiography. PMID- 16252035 TI - Not-so-obvious considerations for professional liability insurance. PMID- 16252037 TI - Tributes to Dale Coln, MD, upon his retirement. PMID- 16252036 TI - Charged to Medicare: how much is too much? PMID- 16252038 TI - Tributes to Maurice Adam, MD. PMID- 16252039 TI - Facts and ideas from anywhere. PMID- 16252040 TI - Highly stable CuO incorporated TiO(2) catalyst for photo-catalytic hydrogen production from H(2)O. AB - A CuO incorporated TiO(2) catalyst was found to be an active photo-catalyst for the reduction of H(2)O under sacrificial conditions. The catalytic activity originates from the photogeneration of excited electrons in the conduction bands of both TiO(2) and CuO resulting in a build-up of excess electrons in the conduction band of CuO. Consequently, the accumulation of excess electrons in CuO causes a negative shift in the Fermi level of CuO. The efficient inter-particle charge transfer leads to a higher catalytic activity and the formation of highly reduced states of TiO(2)/CuO, which are stable even under oxygen saturated condition. Negative shift in the Fermi level of CuO of the catalyst TiO(2)/CuO gains the required over-voltage necessary for efficient water reduction reaction. The function of CuO is to help the charge separation and to act as a water reduction site. The amount of CuO and crystalline structure were found to be crucial for the catalytic activity and the optimum CuO loading was ca. approximately 5-10%(w/w). PMID- 16252041 TI - Photoisomerization and photo-hydration of 3-hydroxystyrylnaphthalenes. AB - Fluorescence and trans-->cis photoisomerization are the main deactivation paths following excitation of trans-1-(2'-naphthyl)-2-(3'-hydroxyphenyl)ethene (trans 2,3NOH) in cyclohexane, methanol and acetonitrile. The quantum yield of both processes is wavelength dependent: this is due to the presence of conformational isomers deriving from rotation of the naphthyl group around the single bond with ethene. Addition of water to acetonitrile quenches the fluorescence (lambda(max)= 380 nm). In CH(3)CN/H(2)O (4/6, v/v) the emission spectrum displays a broad band with maximum at approximately 550 nm besides the original quenched fluorescence. This indicates that 2,3-NOH undergoes acid-base equilibration in the singlet excited state as supported by the enhancement of the fluorescence quantum yield with increasing acidity of the medium. Ground and excited state acidity constants have been determined. The main photochemical process is photo-hydration, i.e. water addition to the ethene bond. Fluorescence and photo-hydration have the same sigmoidal dependence on the acid concentration, which indicates that the undissociated form of singlet excited 2,3NOH is the photoreactive species. Laser flash photolysis experiments allowed identification of the reactive intermediates. The photophysics of trans-1-(1'-naphthyl)-2-(3' hydroxyphenyl)ethene (trans-1,3NOH) is similar to that of 2,3NOH as regards the effect of water on fluorescence and the acid-base behaviour in the ground and first excited singlet state; the main photochemical process is trans-->cis photoisomerization together with photo-cylization to hydroxychrysene in neutral water/acetonitrile, but with lower yield compared to cyclohexane, and photo hydration in strongly acidic medium. PMID- 16252042 TI - Time evolution and competing pathways in photodegradation of trifluralin and three of its major degradation products. AB - The herbicide trifluralin (I)(N,N-di-n-propyl-2,6-dinitro-4 trifluoromethylaniline) decomposes, by the action of UV-Vis light (lambda > 300 nm), to several products, the most important (because they give subsequent photochemical reactions) being N-n-propyl-2,6-dinitro-4-trifluoromethylaniline (VI), 2-ethyl-7-nitro-5-trifluoromethyl-1H-benzimidazole 3-oxide (VII) and 2,6 dinitro-4-trifluoromethylaniline (XII). The time evolution of degradation of trifluralin (I) and the aforementioned three main photoproducts was studied in water and acetonitrile as solvents. The pseudo-first order rate constants allow one to calculate the branching ratios for some of the reactions involved. The preference for either N-dealkylation or cyclization depends on the solvent employed. Dissolved oxygen accelerates the photodegradation, especially the dealkylation. PMID- 16252043 TI - Photochemical rearrangement of dibenzo[1,4]dioxins proceeds through reactive spirocyclohexadienone and biphenylquinone intermediates. AB - Photochemical studies on a range of model dibenzo[1,4]dioxins were performed in aqueous and organic solutions. The compounds were found to undergo a photochemically initiated aryl-ether bond homolysis that yields reactive 2-spiro 6'-cyclohexa-2',4'-dien-1'-one and subsequent 2,2'-biphenylquinone intermediates. Under steady-state irradiation, the 2,2'-biphenylquinones were observed to participate in excited state hydrogen abstraction from the organic solvent to give the corresponding 2,2'-dihydroxybiphenyls. In the absence of continued irradiation, 2,2'-biphenylquinones with electron donating substituents thermally rearrange to the corresponding oxepino[2,3-b]benzofurans, whereas the unsubstituted 2,2'-biphenylquinone and its derivatives with electron withdrawing groups thermally rearrange to the corresponding 1-hydroxydibenzofurans. PMID- 16252044 TI - Synthetic and photochemical studies of substituted 1-acyl-7-nitroindolines. AB - A previous study of substituent effects on the photo-cleavage of 1-acyl-7 nitroindolines has been extended to examine the effects of electron-donating and electron-withdrawing substituents. 1-Acetyl-4,5-methylenedioxy-7-nitroindoline was inert to 350 nm irradiation, reinforcing an earlier finding that excessive electron-donation by substituents can divert the excited state into non productive pathways. By contrast, the 1-acetyl-5,7-dinitro- and 1-acetyl-4 methoxy-5,7-dinitroindolines and respectively both showed improved photolysis efficiency in aqueous solution compared to the 1-acyl-4-methoxy-7-nitro compound . Unlike , both and gave mixed photoproducts, the corresponding dinitroindolines and the 5-nitro-7-nitrosoindoles. These results are interpreted in terms of a previous mechanistic study. Investigation of the 4-methoxy-5,7-dinitroindoline conjugate of L-glutamate showed that the stoichiometry of glutamate release upon photolysis was only 65-77% of the theoretical value, suggesting that photolysis of these dinitro compounds may involve pathways other than the clean photolysis previously observed for mono-nitro compounds such as . PMID- 16252045 TI - Intermediates formed by laser flash photolysis of [PtCl(6)](2-) in aqueous solutions. AB - The stationary photolysis of [PtCl(6)](2-) in aqueous solutions (10(-5)-10(-4) M) at the region of 313 nm leads to its photoaquation with a quantum yield of 0.19. Laser flash photolysis experiments (308 nm) provided evidence of the formation of Pt(iii) intermediates, namely [PtCl(4)(OH)(H(2)O)](2-) and [PtCl(4)](-), and Cl(2) (-) radical anions. The Pt(iii) complexes formed as a result of an intrasphere electron transfer from Cl(-) ligands to the excited Pt(iv) ion. However, the main ( approximately 90%) photolysis channel was not accompanied by the transfer of Cl atoms to the solvent bulk. The photoaquation of [PtCl(6)](2-) results from the back electron transfer in the secondary geminate pair, [PtCl(5)(H(2)O)](2-)-Cl. The relative yield of Pt(iii) intermediates, recorded after the completion of all processes in the geminate pair, was less than 10% of the number of disappearing initial [PtCl(6)](2-) complexes. PMID- 16252046 TI - Nonlinear photodynamic therapy: method of pulsed oxygen depletion. AB - The photodynamic therapy technique involving pulsed oxygen depletion (POD) in tissue by long high-energy pulses of light was studied theoretically. The possibility of creating a uniform distribution of a therapeutic dose throughout a tumor using both surface and interstitial irradiation is shown. Possible thickness of the treated tissue layer is estimated. The comparison with other methods of nonlinear photodynamic therapy is made. PMID- 16252047 TI - Potential of phenothiazine as a thin film dosimeter for UVA exposures. AB - The research reported in this paper on the changes in absorbance and the calibration of a proposed UVA (320-400 nm) dosimeter have established the phenothiazine-mylar combination as a potential UVA dosimeter for population studies of UVA exposures. The change in optical absorbance at 370 nm was employed to quantify the UVA exposures. This change starts to saturate at a change in absorbance of approximately 0.3. This relates to solar UVA exposures at a sub tropical site on a horizontal plane of approximately three to four hours. The shape of this calibration curve varies with the season. This can be overcome in the same manner as for polysulfone where the dosimeter is calibrated for the conditions that it will be employed to measure the UVA exposures. PMID- 16252048 TI - Light distribution and calibration of commercial PDT LED arrays. AB - Commercial photodynamic therapy (PDT) light sources are supplied with no evidence of traceability to national measurements standards to validate indicated delivered dose. Also, the spatial distribution of the radiant energy is not disclosed. This means that there is no way for the user to be able to verify that the required dose is being delivered to the appropriate area of the lesion. To this end, a simple method, traceable to national standards, is described and applied to an investigation of two commercial LED arrays. In one case, the dose fell to 38% of that received at the central area at a distance of only 2 cm. In the other, the output was more uniform over a larger area but the maximum irradiance was not at the centre of the field. All of these inhomogenieties were taken into account when the actual light dose delivered to the patient was calculated in the method described. This ensured transparent traceability in PDT dosimetry. PMID- 16252051 TI - Health-related aerosol measurement: a review of existing sampling criteria and proposals for new ones. AB - Interest in particle size-selective sampling for aerosols in working and ambient living environments began in the early 1900s when it became apparent that the penetration into-and deposition in-the respiratory tract of aerosol-exposed humans of inhaled particles was dependent on particle size. Coarse particles tended to be filtered out during inhalation and in the upper parts of the respiratory tract, so only progressively smaller particles penetrated down to the deep regions of the lung. Over time, following experimental studies with 'breathing' mannequins in wind tunnels and with human volunteer subjects in the laboratory, a clear picture has emerged of the physical, physiological and anatomical factors that control the extent to which particles may or may not reach certain parts of the respiratory tract. Such understanding has increasingly been the subject of discussions about aerosol standards, in particular the criteria by which exposure might be defined in relation to given classes of aerosol-related health effect-and in to turn aerosol monitoring. The ultimate goal has been to develop a set of criteria by which exposure standards are scientifically relevant to the health effects in question. This paper reviews the scientific basis for such criteria. It discusses the criteria that have already been widely discussed and so are either being applied or are on the threshold of practical application in standards. It also discusses how new advanced knowledge may allow us to extend the list of particle size-selective criteria to fractions that have not yet been widely discussed but which may be of importance in the future. PMID- 16252052 TI - Injectors for capillary gas chromatography and their application to environmental analysis. AB - The application of different injectors in capillary gas chromatography (GC) analysis of semi-volatile organic contaminants in environmental samples prepared in organic solvents is reviewed. The injectors examined include a split/splitless injector in splitless mode (SS), cold on-column (COC), and programmable temperature vaporizer (PTV) and adaptations of these injector designs. Key issues when selecting an injector include properties of the analyte, such as potential for thermal degradation or discrimination of high boiling point compounds within the injector, and the ability of the GC systems to handle large volume injections (LVI) primarily to lower detection limits and allow direct coupling with sample preparation techniques such as at-line or on-line solid phase extraction (SPE). LVI also require consideration of matrix interference issues. This review examines only injector chamber modifications that are feasible with a standard GC configuration, however some modifications to the chromatographic system to extend the range of applicability of gas chromatography analysis for environmental samples are also noted. PMID- 16252053 TI - Carbon speciation in airborne particulate matter with C (1s) NEXAFS spectroscopy. AB - Recent and current research activities on the chemical characterization of carbon in airborne carbonaceous particulate matter with near-edge X-ray absorption fine structure (NEXAFS) spectroscopy are reviewed. NEXAFS spectroscopy uses soft X rays from synchrotron radiation facilities and allows for the bulk and surface speciation of particulates smaller than 2.5 micrometres (PM 2.5). This relatively novel technique is often superior to TEM-EELS and FTIR spectroscopy. In the extreme case, one single PM particle is sufficient for characterization. Liquids, extracts, solid core and surface functional groups can be quantified. Preliminary data on combustion derived PM such as diesel soot, wood smoke and tobacco smoke are compared with ambient samples. PMID- 16252054 TI - Municipal landfills exhale newly formed organotins. AB - For the first time we are able to report the identification and quantification of several unexpected alkylated tin compounds such as dimethyldiethyltin, trimethylethyltin and propyltrimethyltin in European municipal waste deposits, by using GC-ICP-MS. Future studies will reveal whether their origin is from the degradation of butyl-, or octyltin compounds or simply products of de novo synthesis within the landfill environment. PMID- 16252055 TI - The study of age influence on human bone lead metabolism by using a simplified model and X-ray fluorescence data. AB - Long term lead metabolism in the human body has never been fully understood due to the lack of human data in this area. The technological improvement of bone lead measurement systems has made bone lead data of substantial populations available. In this study, a set of X-ray fluorescence bone lead data was used to test Leggett's lead metabolism model (R. W. Leggett, Environ. Health Perspect., 1993a, 101, 598-616), especially the model of metabolism in bone. The data set includes the bone lead concentration of 539 occupationally exposed workers, of whom 327 were measured twice in five years. The bone lead concentrations of both cortical bone (tibia) and trabecular bone (calcaneus) were obtained by Cd-109 gamma-ray induced XRF measurement. The histories of blood lead concentration for these workers were used to regulate the input file of the model. The results show that the bone lead concentrations predicted by Leggett's model greatly underestimate the measured values, especially for older workers. This data set was then organized into five age groups. A new simplified model was applied to estimate the lead transfer rates between blood and lead compartments for these age groups. The original transfer rates and the new transfer rates are compared, and the differences are discussed. When the transfer rates derived from measured bone lead data were put into the input file of the model to replace the existing parameters, the predicted values were much closer to the measured values for both cortical bone and trabecular bone. PMID- 16252056 TI - Occurrence of perfluorosulfonates and other perfluorochemicals in dust from selected homes in the city of Ottawa, Canada. AB - A series of perfluorinated compounds (PFCs) including perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA) have been recently measured in a variety of environmental samples and biological matrices. In order to better understand the human exposure routes of these chemicals, levels of PFOS, PFOA, perfluorobutane sulfonate (PFBS), perfluorohexane sulfonate (PFHS) and perfluorooctane sulfonamide (PFOSA) in house dust samples were investigated. The data revealed a correlation between the concentrations of PFCs and the percentage of carpeting in the house; older houses tended to have less carpeting, hence lower levels of these perfluorinated compounds in their dust. PMID- 16252057 TI - Investigation of arsenic speciation on drinking water treatment media utilizing automated sequential continuous flow extraction with IC-ICP-MS detection. AB - Three treatment media, used for the removal of arsenic from drinking water, were sequentially extracted using 10 mM MgCl2(pH 8), 10 mM NaH2PO4(pH 7) followed by 10 mM (NH4)2C2O4(pH 3). The media were extracted using an on-line automated continuous extraction system which allowed the arsenic in each of the extraction fluids to be speciated on-line using IC-ICP-MS. The 10 mM MgCl2 preferentially extracted As(III) from each of the media. The percentage of the arsenic extracted by the MgCl2, relative to a HNO3/H2O2 digestion of the media, ranged from 0.1 2.3% for the three solids. The next sequential extraction fluid, 10 mM NaH2PO4, extracted some of the residual As(III) remaining on each of the media but the predominant species extracted was As(V). The 10 mM NaH2PO4 extracted 15.3 to 42.8% of the total arsenic relative to a total digested concentration for each of the media. The As(III) and As(V) stability studies conducted in these two extraction fluids indicated that conversion between As(III) and As(V) was not significant for the short extraction fluid sample contact time associated with the on-line continuous flow extraction cell. Finally, the 10 mM (NH4)2C2O4 extraction fluid was utilized in an off-line analysis mode because the Fe and As concentrations extracted from the media were not compatible with direct ICP-MS detection. The (NH4)2C2O4 extracted 2.9-29% As(III) for all three media and caused an oxidation of As(III) to As(V) during the extraction period for one of the three media. The sum of the arsenic from each of the three extraction fluids represented 92%, 44% and 53% of the available total arsenic for the three media, respectively. The speciation results for each media were obtained by adding all the speciation results from all three extraction fluids together and the resulting distribution of As(III)/As(V) compared well with the speciation results obtained via XANES. PMID- 16252058 TI - Optimization of fertirrigation efficiency in strawberry crops by application of fuzzy logic techniques. AB - A high level of price support has favoured intensive agriculture and an increasing use of fertilisers and pesticides. This has resulted in the pollution of water and soils and damage to certain eco-systems. The target relationship that must be established between agriculture and environment can be called "sustainable agriculture". In this work we aim at relating strawberry total yield with nitrate concentration in water at different soil depths. To achieve this objective, we have used the Predictive Fuzzy Rules Generator (PreFuRGe) tool, based on fuzzy logic and data mining, by means of which the dose that allows a balance between yield and environmental damage minimization can be determined. This determination is quite simple and is done directly from the obtained charts. This technique can be used in other types of crops permitting one to determine in a precise way at which depth the appropriate dose of nitrate fertilizer must be correctly applied, on the one hand providing the maximum yield but, on the other hand, with the minimum loss of nitrates that leachate through the saturated zone polluting aquifers. PMID- 16252059 TI - Uptake and bioavailability of persistent organic pollutants by plants grown in contaminated soil. AB - This paper assesses the uptake of persistent organic pollutants (POP's) into plants. In particular, uptake of alpha-endosulfan, beta-endosulfan and endosulfan sulfate from lettuce. The lettuce plants were grown on compost that had previously been contaminated at 10 and 50 microg g(-1) per POP. The soil was slurry spiked by adding the appropriate amount of POP in acetone in an approximate ratio of 1 ratio 2, w/v soil ratio solvent. The solvent was left to evaporate at ambient temperature for 24 hours. Lettuce plants were grown under artificial daylight for 12 hours a day. The influence of soil ageing on the recovery of POP's from spiked soil samples was also assessed. The average recovery of endosulfan compounds from slurry spiked soil (10, 20 and 40 microg g( 1)) was consistent (92.9 +/- 4.4% for n= 9). However, ageing of endosulfan compounds on the slurry spiked soil resulted in lower recoveries (average losses were 12.5% after 14 days ageing of slurry spiked soil). The uptake of POP's was assessed by measuring the amount of endosulfan compounds in roots and leaves from lettuce plants after 10, 20 and 33 days. In addition, control plants grown in uncontaminated soil were monitored and analysed. It was found that endosulfan compounds were present in the roots of all lettuce plants irrespective of soil spike level or age of plant. In the 33 day lettuce plants where the soil was spiked at the highest level (50 microg g(-1)) endosulfan compounds were determined in the leaves. The root to leaf ratio was found to be 3.1 for alpha endosulfan, 46.0 for beta-endosulfan, and 24.3 for endosulfan sulfate. Spiked lettuce samples were subjected to in vitro gastrointestinal extraction to assess the bioavailability of endosulfan compounds. No detectable endosulfan compounds were determined in the gastric extracts while small quantities (range 0.06-0.12 microg g(-1)) were found in the intestinal extraction. All samples (soil and lettuce) were extracted using pressurised fluid extraction and analysed using gas chromatography with mass selective detection. PMID- 16252061 TI - Comment on "model for the adsorption of organic compounds at gas-water interfaces" by C. F. Poole, JEM, 2005, 7, 577. PMID- 16252060 TI - Comparison of field-observed and model-predicted plume trends at fuel contaminated sites: implications for natural attenuation rates. AB - Subsequent to modeling of natural attenuation processes to predict contaminant trends and plume dynamics, monitoring data were used to evaluate the effectiveness of natural attenuation at reducing contaminant concentrations in groundwater at seven fuel-contaminated sites. Predicted and observed contaminant trends at seven sites were compared in order to empirically assess the accuracy of some fundamental model input parameters and assumptions. Most of the models developed for the study sites tended to overestimate plume migration distance, source persistence, and/or the time required for the benzene, toluene, ethyl benzene, and xylenes (BTEX) plumes to attenuate. Discrepancies between observed and predicted contaminant trends and plume behavior suggested that the influence of natural attenuation process may not have been accurately simulated. The conservatism of model simulations may be attributed to underestimation of natural source weathering rates, overestimation of the mass of contaminant present in the source area, and/or use of overly conservative first-order solute decay rates. PMID- 16252063 TI - Analysis of the NSD1 promoter region in patients with a Sotos syndrome phenotype. AB - Sotos syndrome (SoS, OMIM#117550) is an overgrowth disorder characterized by excessive growth-especially in the first years of childhood-distinctive craniofacial features, and various degrees of mental retardation. Haploinsufficiency of the nuclear receptor binding SET domain containing protein 1 (NSD1) gene, due to either intragenic mutations or whole-gene microdeletions, is found in the majority of patients with SoS. However, in approximately 10-40% of patients with a typical SoS phenotype, no abnormalities are detected. In this study, hemizygous hypermethylation or genomic sequence abnormalities of the promoter region of NSD1 were hypothesized to be the underlying cause in patients with a SoS phenotype, but without confirmed NSD1 alterations. In 18 patients, including one patient with a reported hepatocellular carcinoma, the promoter region of NSD1 was analyzed. However, no hypermethylation or sequence abnormalities in the promoter region could be detected. It therefore seems unlikely that such abnormalities of NSD1 are a major culprit in patients with phenotypical SoS. Additional methods are necessary for detection of other genetic or epigenetic causes of SoS. PMID- 16252064 TI - Basal ganglia volumetric studies in affective disorder: what did we learn in the last 15 years? AB - Until today, morphometric neuroimaging studies on affective disorders concentrate on the limbic system, especially the hippocampus, amygdala, and anterior cingulate. In most of the studies and reviews available today, the basal ganglia are of secondary interest. It seems that the basal ganglia are interest of neurologist, whereas the limbic system is reserved for psychiatric neuroimaging studies. We follow a different approach in this review, studying all available papers on MRI research of the basal ganglia in unipolar depression and bipolar disorder. We found a possibly larger neostriatum in bipolar and possibly smaller one in unipolar patients. None of the unipolar studies found any larger basal ganglion, and only one out of 12 bipolar studies found smaller basal ganglia. Both findings seemed to depend on age (tendency toward smaller volumes in unipolar and bipolar with older age), sex (men tending to pathology in both disorders) and bipolar patients show a possible influence of medication, which is not assessed so far in unipolar depression. We conclude that several methodological shortcomings in volumetric MRI research on the basal ganglia in affective disorders make it necessary to imply more research in this area. We suggest (a) better MRI methods (we do not have a single volumetric 3 Tesla study in this patient group); (b) studies of medication-naive patients (thus ruling out the medication effect); (c) Studies that directly compare unipolar depressed and bipolar patients are needed to determine whether these apparent differences in morphometric abnormalities, as observed through the mediating comparison with healthy subjects, are real. PMID- 16252065 TI - Synergistic effect of alpha-dihydroergocryptine and L-dopa or dopamine on dopaminergic neurons in primary culture. AB - There is an ongoing controversy about potential toxicity of L-3,4 dihydroxyphenylalanine (L-dopa) to dopaminergic neurons in Parkinson's disease (PD). Neuroimaging data suggest that L-dopa accelerates the loss of dopamine nerve terminals, especially at higher doses. The disputed aspect of toxicity and the frequently observed motor complications accompanying L-dopa therapy have led to an increased use of dopamine agonists during the past two decades. Reports describing their neuroprotective potential to dopaminergic neurons have attracted much attention. Here, we describe the novel finding that the combination of a dopamine (DA) agonist, alpha-dihydroergocryptine (DHEC), with L-dopa or DA exerts a synergistic stimulatory effect on dopaminergic neurons in primary culture, while each substance alone had no or less effect. DA receptor stimulation plays a decisive role. The synergistic effect suggests that a combinatory therapy can be beneficial to slow the degeneration of dopaminergic neurons. PMID- 16252066 TI - Serotonergic mechanisms in Parkinson's disease: opposing results from preclinical and clinical data. AB - Parkinson's disease (PD) is a neuropsychiatric disease affecting approximately 1 2% of the general population. The classical triad of symptoms, tremor, rigidity, and bradykinesia is mainly caused by degeneration of dopaminergic neurons from the substantia nigra. However, other neurotransmitter systems also show signs of degeneration, among which the serotonergic system. The exact role of serotonin in PD remains unclear. We present here a review about functional serotonergic interventions and serotonergic imaging studies in PD, and will go into the importance of combining preclinical and clinical research data in order to gain more insight into the role of serotonin in PD. More specifically, the present review is aimed at bridging the gap between data from animal models of PD and data from human research. PMID- 16252067 TI - Cyclosporine A (CsA) affects the pharmacodynamics and pharmacokinetics of the atypical antipsychotic amisulpride probably via inhibition of P-glycoprotein (P gp). AB - The importance of P-glycoprotein (P-gp) in the pharmacokinetics of amisulpride and the effects of a P-gp inhibitor cyclosporine A (CsA) was investigated both, in vitro and in vivo. In vitro and in vivo results indicated amisulpride as a substrate of P-gp. Amisulpride was not metabolized by rat liver microsomes. Open field behavior showed time dependent abolishment in locomotion by amisulpride (50 mg kg(-1)). Co-administration of CsA (50 mg kg(-1)) resulted in a higher and significantly longer antipsychotic effect (24 h after drug administration). Accordingly, the area under concentration-time curve in serum and brain was higher in CsA co-treated rats (13.5 vs. 29.8 micromol h l(-1) for serum and 2.16 vs 2.98 micromol h l(-1) for brain tissue) while renal clearance was not affected. These results pointed to a pharmacokinetic drug interaction between CsA and amisulpride most likely caused by inhibition of P-gp. PMID- 16252068 TI - DBH*444G/A polymorphism of the dopamine-beta-hydroxylase gene is associated with alcoholism but not with severe alcohol withdrawal symptoms. AB - As the enzyme dopamine-beta-hydroxylase (DbetaH) converts dopamine to norepinephrine and both transmitters seem to be involved in the pathology of alcoholism and severe alcohol withdrawal symptoms, the gene encoding DbetaH (DBH) was applied to explore the genetic background of alcoholism and severe withdrawal symptoms. 102 healthy control subjects and 208 alcoholics, including 97 patients with a history of mild withdrawal symptoms, 57 with a history of alcohol withdrawal seizure (AWS) and 82 with a history of delirium tremens (DT) were genotyped for the DBH*444G/A polymorphism revealing a significantly elevated frequency of genotypes carrying the A-allele (p = 0.02; after Bonferroni adjustment for multiple tests) in alcoholics compared to healthy controls. Frequencies of alleles and genotypes of individuals with mild withdrawal symptoms did not differ significantly from those of patients with DT or AWS. PMID- 16252069 TI - Skeletal muscle HSP72 and norepinephrine response to static magnetic field in rat. AB - The present work was undertaken in order to investigate the noradrenergic system and skeletal muscle heat shock protein 72 (HSP72) response to static magnetic field (MF) in male rats. At thermoneutrality (25 degrees C), the exposition of rats 1 hour/day for 5 consecutive days to MF of 128 mT (m tesla) induced an increase in norepinephrine content in gastrocnemius muscle (+25%, p < 0.05) but had no effect at 67 mT (+1%, p > 0.05), indicating a stimulatory effect of sub acute MF exposure on the noradrenergic system activity. Moreover, exposed rats to MF displayed a non-significant increase of HSP72 levels in gastrocnemius muscles (+29%, p > 0.05). The results indicate that noradrenergic systems in rat's gastrocnemius muscles are affected by MF exposure. Interestingly, sub-acute exposure insufficiency increased HSP72 levels in gastrocnemius muscles. PMID- 16252071 TI - Chronic alcohol intoxication in rats leads to a strong but transient increase in NGF levels in distinct brain regions. AB - Nerve growth factor (NGF), a member of the neurotrophin family, is an essential mediator of neuronal activity and synaptic plasticity of basal forebrain cholinergic neurons. In this study NGF-protein levels were determined in areas of the basal forebrain cholinergic system, its projection areas as well as the striatum and the cerebellum after long-term exposure (6 and 9 months) to ethanol and a phase of withdrawal in male Sprague-Dawley rats. 6-month alcohol treatment led to an increase of NGF to 650-850% of controls in the basal forebrain and the septum and to a 210-485% increase in the cholinergic projection areas (anterior cortex, hippocampus and olfactory bulb). After 9 months exposure to ethanol, a decrease of NGF by 16% in the frontal cortex was observed compared to controls. In the other brain regions no differences in NGF expression were detectable at this time-point. These results support the idea of an endogenous neuroprotective mechanism acting through a transient NGF induction followed by a decrease in NGF levels during the course of further neuronal degeneration. PMID- 16252072 TI - Compulsive exercise acutely upregulates rat hippocampal brain-derived neurotrophic factor. AB - This study was to examine the effects of treadmill exercise on the expression of brain-derived neurotrophic factor (BDNF) in rat hippocampus. After 1-wk treadmill familiarization, animals in exercise groups received a 4-wk exercise training or an acute exercise. They were sacrificed 2 h or 2 d after exercise and their hippocampal BDNF mRNA and protein levels were determined. We demonstrated that 1) hippocampal BDNF mRNA and protein levels were both elevated in response to exercise training at 2 h after the last run but not after 2 d; 2) an acute moderate exercise (1 or 3 d) increased BDNF protein levels; 3) acute severe exercise increased BDNF protein and mRNA levels in animals under a familiarization regimen, while suppressed the BDNF mRNA level in rats without treadmill familiarization, paralleling the stress effect of immobilization/water exposure. We conclude that compulsive treadmill exercise with pre-familiarization acutely upregulates rat hippocampal BDNF gene expression. PMID- 16252070 TI - Testing models of thalamic dysfunction in schizophrenia using neuroimaging. AB - Neural models of schizophrenia have implicated the thalamus in deficits of early sensory processing and multimodal integration. We have reviewed the existing neuroimaging literature for evidence in support of models that propose abnormalities of thalamic relay nuclei, the mediodorsal thalamic nucleus, and large-scale cortico-thalamic networks. Thalamic volume reduction was found in some but not all studies. Studies of the early stages of schizophrenia suggest that thalamic volume reduction is present early in the course of the illness. Functional imaging studies have revealed task related abnormalities in several cortical and subcortical areas including the thalamus, suggesting a disruption of distributed thalamocortical networks. Chemical imaging studies have provided evidence for a loss of thalamic neuronal integrity in schizophrenia. There is, at present, inadequate data to support the hypothesis that schizophrenia is associated with abnormalities of sensory relay or association nuclei. There is evidence for a perturbation of cortico-thalamic networks, but further research is needed to elucidate the underlying mechanisms at the cellular and systems levels. The challenges ahead include better delineation of thalamic structure and function in vivo, the combination of genetic and imaging techniques to elucidate the genetic contributions to a thalamic phenotype of schizophrenia, and longitudinal studies of thalamic structure and function. PMID- 16252073 TI - Interaction analysis between 5-HTTLPR and TNFA -238/-308 polymorphisms in schizophrenia. AB - This study investigated the potential interaction between the polymorphisms of serotonin transporter gene (SLC6A4, a 44 base pair insertion/deletion in the promoter region, 5-HTTLPR) and tumor necrosis factor-alpha gene (TNFA; -238G/A and -308G/A polymorphisms) on the development of schizophrenia, as well as the interaction of the three polymorphisms in relation to symptomatology, family history, onset age and antipsychotic treatment response. Genomic DNA analyses with polymerase chain reaction (PCR) was used for the genotyping. One hundred and fifty-two (152) patients with schizophrenia and 152 normal controls participated in the study. Any associations between the individual polymorphism and schizophrenia were not found. However, marginal association between subjects with both TNFA -238 A allele (genotype AA plus AG) and 5-HTTLPR s allele (ss plus sl) and presence of family history was found (p = 0.023; p = 0.026). The subjects with TNFA -308 AG genotype showed higher change in PANSS total score (p = 0.028). No significant interaction effect between 5-HTTLPR and TNFA -238/-308 polymorphisms either on the development of schizophrenia or on antipsychotics treatment response and psychopathology was found, although a significant interaction effect for subjects carrying TNFA -238 AG and -308 AA genotypes on a positive family history was observed (p = 0.017). These results suggest that the interaction effects between 5-HTTLPR and TNFA -238/-308 polymorphisms gives no significant contribution to the susceptibility to schizophrenia, and is not associated with clinical variables, antipsychotic treatment response and psychopathological features, except for family history of disease, at least in the Korean population. PMID- 16252074 TI - Evidence for a neurophysiologic auditory deficit in children with benign epilepsy with centro-temporal spikes. AB - Benign focal epilepsy in childhood with centro-temporal spikes (BECTS) is one of the most common forms of epilepsy. Recent studies have questioned the benign nature of BECTS, as they have revealed neuropsychological deficits in many domains including language. The aim of this study was to investigate whether the epileptic discharges during the night have long-term effects on auditory processing, as reflected on electrophysiological measures, during the day, which could underline the language deficits. In order to address these questions we recorded base line electroencephalograms (EEG), sleep EEG and auditory event related potentials in 12 children with BECTS and in age- and gender-matched controls. In the children with BECTS, 5 had unilateral and 3 had bilateral spikes. In the 5 patients with unilateral spikes present during sleep, an asymmetry of the auditory event related component (P85-120) was observed contralateral to the side of epileptiform activity compared to the normal symmetrical vertex distribution that was noted in all controls and in 3 the children with bilateral spikes. In all patients the peak to peak amplitude of this event related potential component was statistically greater compared to the controls. Analysis of subtraction waveforms (deviant - standard) revealed no evidence of a mismatch negativity component in any of the children with BECTS. We propose that the abnormality of P85-120 and the absence of mismatch negativity during wake recordings in this group may arise in response to the long-term effects of spikes occurring during sleep, resulting in disruption of the evolution and maintenance of echoic memory traces. These results may indicate that patients with BECTS have abnormal processing of auditory information at a sensory level ipsilateral to the hemisphere evoking spikes during sleep. PMID- 16252075 TI - Lack of glutathione peroxidase-1 exacerbates Abeta-mediated neurotoxicity in cortical neurons. AB - The aetiologies of Alzheimer's disease (AD) are complex and multifactorial. Current therapies are largely ineffective, as the pathophysiological pathways are poorly understood. Observations in AD autopsies, as well as in vivo and in vitro observations in transgenic mice, have implicated oxidative stress as pathogenic in AD. This study used the Glutathione Peroxidase-1 knockout mouse (Gpx1--/--) model to investigate the role of antioxidant disparity in neuropathologies. Cultured neurons from control and Gpx1--/-- embryos were treated with AD-related peptides and the degree of cell loss compared. Results show that antioxidant disparity makes Gpx1--/-- cells more susceptible to Abeta toxicity. Surrogate replacement of Gpx1 with the reactive oxygen species scavenger N-acetyl cysteine and the Gpx1 mimetic ebselen, reverses the Gpx1--/-- increased susceptibility to Abeta toxicity. Such results support a role for oxidative stress in AD-related neuronal loss. This study is the first to report such findings using the Gpx1--/- model, and supports a role for oxidative stress as one of the contributing factors, in development of AD-like pathologies. PMID- 16252076 TI - The antioxidant anethole dithiolethione inhibits monoamine oxidase-B but not monoamine oxidase A activity in extracts of cultured astrocytes. AB - Anethole dithiolethione (ADT) is a clinically available, pluripotent antioxidant proposed as a neuroprotectant for Parkinson's disease (PD). Here, using extracts from cultured astrocytes, containing both monoamine oxidase (MAO) A and B activity, we demonstrate that ADT concentration-dependently inhibits MAO-B activity in a clinically relevant concentration range (0.03-30 microM, IC-50 = 0.5 microM) without affecting MAO A activity. Considering the alleged contribution of MAO activity in general, and MAO-B in particular, to oxidative stress and neurodegeneration in PD, our data further support the neuroprotective potential of ADT. PMID- 16252077 TI - Consensus guidelines for validation of virtual reality surgical simulators. AB - The Work Group for Evaluation and Implementation of Simulators and Skills Training Programmes is a newly formed sub-group of the European Association of Endoscopic Surgeons (EAES). This work group undertook a review of validation evidence for surgical simulators and the resulting consensus is presented in this article. Using clinical guidelines criteria, the evidence for validation for six different simulators was rated and subsequently translated to a level of recommendation for each system. The simulators could be divided into two basic types; systems for laparoscopic general surgery and flexible gastrointestinal endoscopy. Selection of simulators for inclusion in this consensus was based on their availability and relatively widespread usage as of July 2004. Whilst level 2 recommendations were achieved for a few systems, it was clear that there was an overall lack of published validation studies with rigorous experimental methodology. Since the consensus meeting, there have been a number of new articles, system upgrades and new devices available. The work group intends to update these consensus guidelines on a regular basis, with the resulting article available on the EAES website (http://www.eaes-eur.org ). PMID- 16252078 TI - Use of non-invasive phase contrast magnetic resonance imaging for estimation of atrial septal defect size and morphology: a comparison with transesophageal echo. AB - BACKGROUND: Transesophageal echocardiography (TEE) is a trusted method of sizing atrial septal defect (ASD) prior to percutaneous closure but is invasive, uncomfortable, and may carry a small risk of morbidity and mortality. Magnetic resonance imaging (MRI) may be useful non-invasive alternative in such patients who refuse or are unable to tolerate TEE and may provide additional information on the shape of the A0SD. PURPOSE: To validate the accuracy of ASD sizing by MRI compared with TEE. METHOD: Twelve patients (mean age 30 years; range 11-60 years) scheduled for ASD closure underwent TEE, cine balanced fast field echo MRI (bFFE MRI) in four-chamber and sagittal views and phase-contrast MRI (PC-MRI) with reconstruction using the two orthogonal planes of T2-weighted images as planning. The average of the three longest measurements for all imaging modalities was calculated for each patient. RESULTS: Mean maximum ASD length on TEE was 18.8 +/- 4.6 mm, mean length by bFFE-MRI was 20.0 +/- 5.0 mm, and mean length by PC-MRI was 18.3 +/- 3.6 mm. The TEE measurement was significantly correlated with the bFFE-MRI and PC-MRI measurements (Pearson r = 0.69, p = 0.02 and r = 0.59, p = 0.04, respectively). The mean difference between TEE and bFFE-MRI measurements was -1.2mm (95% CI: -3.7, 1.3) and between TEE and PC-MRI was 0.5 mm (95% CI: 1.9, 2.9). Bland-Altman analysis also determined general agreement between both MRI methods and TEE. The ASDs were egg-shaped in two cases, circular in 1 patient and oval in the remaining patients. CONCLUSION: ASD sizing by MRI using bFFE and phase-contrast protocols correlated well with TEE estimations. PC-MRI provided additional information on ASD shapes and proximity to adjacent structures. PMID- 16252080 TI - Failed retrieval of potentially retrievable IVC filters: a report of two cases. AB - Retrievable inferior vena cava filters are being increasingly used for the prevention of life-threatening pulmonary emboli in patients who have temporary contraindications to anticoagulation therapy. We report two cases of failure to remove these devices. PMID- 16252079 TI - PTA versus carbofilm-coated stents in infrapopliteal arteries: pilot study. AB - PURPOSE: To determine the primary success and short-term patency of stent application as a primary treatment modality for high-grade lesions of the infrapopliteal arteries compared with treatment with percutaneous transluminal angioplasty (PTA) in critical limb ischemia in a randomized prospective study. METHODS: Endovascular therapy was performed on 95 lesions in 51 patients (mean age 72.0 years, range 47-80 years) who presented clinically with Fontaine stages III and IV. One patient underwent treatment in both limbs. After angiographic lesion identification, patients were randomized for treatment by PTA (53 lesions in 27 patients) or stent application (42 lesions in 24 patients). Follow-up by clinical investigation and conventional angiography or spiral CT angiography was performed in 37 patients (57 lesions) 6 to 12 months after the procedure, or when clinically indicated. Evaluation was performed by two observers, double-blinded, with thresholds for lesion restenosis of 50% and 70%. Statistical evaluation was performed on a lesion basis by Kaplan-Meier estimated probability rates, and log rank and Wilcoxon tests. The primary endpoint was the angiographic patency rate of treated lesions. RESULTS: The inter-reader agreement was high (kappa = 0.82). For the stent group the cumulative primary patency at 6 months was 83.7% at the 70% restenosis threshold, and 79.7% at the 50% restenosis threshold. For PTA, the primary patency at 6 months was 61.1% at the 70% restenosis threshold and 45.6% at the 50% restenosis threshold. Both results were statistically significant (p < 0.05). CONCLUSION: Infrapopliteal stent application is an effective treatment modality for high-grade lesions in chronic critical limb ischemia. Compared with PTA, higher patency rates can be expected after 6 months. PMID- 16252082 TI - Resolution of large azygos vein aneurysm following stent-graft shunt placement in a patient with Ehlers-Danlos syndrome type IV. AB - Ehlers-Danlos syndrome (EDS) type IV is a rare connective tissue disorder associated with thin-walled, friable arteries and veins predisposing patients to aneurysm formation, dissection, fistula formation, and vessel rupture. Azygos vein aneurysm is an extremely rare condition which has not been reported in association with EDS in the literature. We present a patient with EDS type IV and interrupted inferior vena cava (IVC) with azygos continuation who developed an azygos vein aneurysm. In order to decrease flow through the azygos vein and reduce the risk of aneurysm rupture, a stent-graft shunt was created from the right hepatic vein to the azygos vein via a transhepatic, retroperitoneal route. At 6 month follow-up the shunt was open and the azygos vein aneurysm had resolved. PMID- 16252081 TI - Percutaneous trans-hepatic obliteration for bleeding esophagojejunal varices after total gastrectomy and esophagojejunostomy. AB - A 72-year-old man who had undergone a total gastrectomy with a Roux-en-Y esophagojejunostomy for gastric cancer 6 years earlier presented to our hospital with massive hematemesis and melena. Endoscopic examination indicated esophageal varices with cherry-red spots and hemorrhage arising from beyond the anastomosis. Abdominal contrast-enhanced computed tomography and angiography revealed a dilated vein in the elevated jejunal limb supplying the varices. Percutaneous trans-hepatic obliteration (PTO) of the varices through the jejunal vein was performed using microcoils, ethanolamine oleate, and gelatin sponge cubes. Ten days after the procedure, endoscopic examination revealed reduction and thrombosis of the varices. We consider PTO to be an effective alternative method for treating ruptured esophagojejunal varices after total gastrectomy. PMID- 16252083 TI - Polymorphisms of the DNA mismatch repair gene HMSH2 in breast cancer occurence and progression. AB - The response of the cell to DNA damage and its ability to maintain genomic stability by DNA repair are crucial in preventing cancer initiation and progression. Therefore, polymorphism of DNA repair genes may affect the process of carcinogenesis. The importance of genetic variability of the components of mismatch repair (MMR) genes is well documented in colorectal cancer, but little is known about its role in breast cancer. hMSH2 is one of the crucial proteins of MMR. We performed a case-control study to test the association between two polymorphisms in the hMSH2 gene: an A --> G transition at 127 position producing an Asn --> Ser substitution at codon 127 (the Asn127Ser polymorphism) and a G --> A transition at 1032 position resulting in a Gly --> Asp change at codon 322 (the Gly322Asp polymorphism) and breast cancer risk and cancer progression. Genotypes were determined in DNA from peripheral blood lymphocytes of 150 breast cancer patients and 150 age-matched women (controls) by restriction fragment length polymorphism and allele-specific PCR. We did not observe any correlation between studied polymorphisms and breast cancer progression evaluated by node-metastasis, tumor size and Bloom-Richardson grading. A strong association between breast cancer occurrence and the Gly/Gly phenotype of the Gly322Asp polymorphism (odds ratio 8.39; 95% confidence interval 1.44-48.8) was found. Therefore, MMR may play a role in the breast carcinogenesis and the Gly322Asp polymorphism of the hMSH2 gene may be considered as a potential marker in breast cancer. PMID- 16252084 TI - [Palliative options for esophageal carcinoma]. AB - Considering the limited chances of radically curing esophageal carcinoma, most of these patients are candidates for palliative therapy. Priority should be given to rapid relief of dysphagia. Endoscopic implantation of esophageal prostheses is immediately effective in 90% of patients, whereas the onset of relief is slower with any alternative method. Long-term complications necessitate an endoscopic reintervention in 30% of the prosthesis carriers. Endoscopic prosthesis implantation is also the first-choice treatment for esophagotracheal fistulae. Self-expanding stents need only minimized preceding bougienage, which has lower complication rates than conventional plastic prostheses. Endoscopic laser radiation is better tolerated by patients than prosthesis implantation but is effective only in very short stenoses. Endoluminal brachytherapy with (192)iridium can be justified in patients with a survival expectancy of more than 6 months, the onset of its effect being slower but longer lasting. Palliative chemotherapy and radiochemotherapy are indicated when metastatic dissemination dominates the symptoms. PMID- 16252085 TI - Early effects of iodine on DNA synthesis in sulfur mustard-induced skin lesions. AB - Sulfur mustard (SM) is powerful alkylator and highly cytotoxic blisterogen in both humans and animals. This study in male guinea pigs shows that, at an early stage (5 h) after SM exposure, a marked increase occurred in epithelial nuclear vacuolation, epidermal thickening, and dermal acute inflammation. Topical iodine treatment reduced the severity of these parameters. The rate of DNA synthesis expressed by incorporation of bromodeoxyuridine was reduced upon topical treatment with iodine only or SM only by 46 and 72%, respectively. Iodine treatment following SM exposure exerted an effect similar to that of SM only, indicating that DNA synthesis is not directly involved in the mechanism of action of iodine-induced protection. PMID- 16252086 TI - Separation science in perfume analysis. PMID- 16252087 TI - Pyridinium-based ionic liquid matrices can improve the identification of proteins by peptide mass-fingerprint analysis with matrix-assisted laser desorption/ionization mass spectrometry. AB - Matrix-assisted laser desorption/ionization mass spectrometry has become an indispensable tool for identification of proteins by peptide mass-fingerprint analysis. Selection of the matrix, addition of matrix additives, and sample preparation techniques are known to affect the quality of the spectra and hence protein identification. We investigated the effect of pyridine as matrix additive for the commonly used crystalline matrix alpha-cyano-4-hydroxycinnamic acid (CCA), forming a pyridinium based ionic liquid matrix, on the mass spectra of synthetic peptides and tryptic protein digests. Beside the equimolar mixture of CCA and pyridine, the effect of addition of substoichiometric amounts of the base to the acid was tested. Optimum results in terms of signal-to-noise ratios, reduction of chemical noise, and reduced formation of alkali adducts and matrix clusters were observed for the matrix CCA-pyridine in the molar ratio 2:1. The optimized ionic liquid matrix was used for identification of tryptic digests of six model proteins and for identification of a protein extracted from a two dimensional gel with the proteome of the bacterium Corynebacterium glutamicum, and shown to facilitate protein identification, yielding higher scores and increased sequence coverage compared with pure CCA. Thus CCA-Py 2:1 is a potential alternative for identification and characterization of proteins by peptide mass-fingerprint analysis. PMID- 16252088 TI - Feasibility of FLAG-IDA regimen in cases with relapsed/refractory acute leukemia cases. PMID- 16252090 TI - A model wheat cultivar for transformation to improve resistance to Fusarium Head Blight. AB - Fusarium head blight (FHB), caused primarily by Fusarium graminearum, is a major disease problem in wheat (Triticum aestivum). Genetic engineering holds significant potential to enhance FHB resistance in wheat. Due to the requirement of screening for FHB resistance on flowers at anthesis, the number of screens carried out in a year is limited. Our objective was to evaluate the feasibility of using the rapid-maturing dwarf wheat cultivar Apogee as an alternative genotype for transgenic FHB resistance research. Our transformation efficiency (number of transgenic plants/number of embryos) for Apogee was 1.33%. Apogee was also found to exhibit high FHB susceptibility and reached anthesis within 4 weeks. Interestingly, microsatellite marker haplotype analysis of the chromosome 3BS FHB resistant quantitative trait locus (QTL) region indicated that this region maybe deleted in Apogee. Our results indicate that Apogee is particularly well suited for accelerating transgenic FHB resistance research and transgenic wheat research in general. PMID- 16252089 TI - Characterization of rec15, an early meiotic recombination gene in Schizosaccharomyces pombe. AB - In S. pombe strains mutant for rec15 aberrant ascus morphology, reduced spore viability and severe reduction of meiotic recombination was detected. Genetic and cytological analysis identified frequent interruption of meiosis after the first division, and nondisjunction I, as the main segregation errors in the mutant. Chromosome segregation at meiosis I was not random in rec15, suggesting the presence of a backup system for correct segregation of achiasmate chromosomes. The analysis of meiotic progression in time-course experiments revealed that the major meiotic events, such as the onset of premeiotic DNA synthesis, of horse tail nuclear movement, and of the first meiotic division occurred earlier in rec15 than in wild-type. The early onset of meiotic events is a novel observation for an early recombination mutant and implies a function of rec15 protein already at or before DNA synthesis. PMID- 16252091 TI - Agrobacterium-mediated transformation of seedling-derived maize callus. AB - Efficient production of seedling-derived Type I callus was demonstrated for several corn genotypes including commercial inbred lines. Seeds were germinated on MS-based medium containing 10 mg l(-1) picloram and 3 mg l(-1) 6 benzylaminopurine, which induced the development of axillary buds in the area of coleoptilar node. Nodal sections of 7-10-day old seedlings were isolated, split longitudinally, and placed on callus induction medium supplemented with 2.2 mg l( 1) picloram and 0.5 mg l(-1) 2,4-dichlorophenoxyacetic acid. For lines L4 and L9 the frequency of embryogenic callus induction was 38-42% based on calli per split nodal section. Frequency of callus induction from split nodal sections of seeds germinated on media without growth regulators was 0-3%. Seedling-derived callus of five genotypes was used for Agrobacterium-mediated transformation. Two constructs containing the green fluorescence protein gene and genes for either neomycin phosphotransferase II or glyphosate selection were used in transformation experiments. Transformation frequency varied from 2 to 11% and about 60% of the T(0) plants had 1-2 copies of transgenes. PMID- 16252093 TI - Functional structure of the cell nucleus. PMID- 16252092 TI - Type II topoisomerase activities in both the G1 and G2/M phases of the dinoflagellate cell cycle. AB - Dinoflagellate genomes are large (up to 200 pg) and are encoded in histoneless chromosomes that are quasi-permanently condensed. This unique combination of chromosomal characteristics presents additional topological and cell cycle control problems for a eukaryotic cell, potentially exhibiting novel regulatory requirements of topoisomerase II. The heterotrophic dinoflagellate Crypthecodinium cohnii was used in this study. The topoisomerase II activities throughout its cell cycle were investigated by DNA flow cytometry following enzyme deactivation. Fluorescence microscopy was also used for studying the chromosome morphology of the treated cells. Two classes of topoisomerase II inhibitors were applied in our study, both of which caused G1 delay as well as G2/M arrest in the C. cohnii cell cycle. At high doses, the topoisomerase poisons amsacrine and ellipticine induced DNA fragmentation in C. cohnii cells. Topoisomerase II activities, as measured by the ability to decatenate kinetoplastid DNA (kDNA), are normally detected throughout the cell cycle in C. cohnii. Our results suggest that the requirement of type II topoisomerase activities during the G1 phase of the cell cycle may relate to the unwinding of quasi-permanently condensed chromosomes for the purpose of transcription. This was also the first time that topoisomerase II activity in dinoflagellate cells was detected. PMID- 16252094 TI - Expression of cysteine sulfinate decarboxylase (CSD) in male reproductive organs of mice. AB - Cysteine sulfinate decarboxylase (CSD) is the rate-limiting biosynthetic enzyme of taurine, but it is still controversial whether the male reproductive organs have the function to synthesize taurine through CSD pathway. The present study was thus undertaken to detect CSD expression in male mouse reproductive organs by RT-PCR, Western blot and immunohistochemistry. The results show that CSD is expressed both at the mRNA and protein levels in the testis, epididymis and ductus deferens. The relative levels of both CSD mRNA and protein increase from the testis to the epididymis and to the ductus deferens. Immunohistochemical results demonstrate that the main cell types containing CSD are Leydig cells of testis, epithelial cells and some stromal cells throughout the efferent ducts, epididymis and ductus deferens. These results suggest that male genital organs have the function to produce taurine through the CSD pathway, although quantifying the relation of CSD expression to taurine synthesis and the exact functions of taurine in male genital organs still need to be elucidated in future studies. PMID- 16252096 TI - Paraoxonase, anti-oxidant response and oxidative stress in children with chronic renal failure. AB - Increased oxidative stress is believed to contribute to an increased risk of cardiovascular disease in uraemia. In children with chronic renal failure (CRF), an anti-oxidant enzyme, paraoxonase (PON), that inhibits oxidation of LDL cholesterol, has not been previously investigated. In this study we aimed to investigate PON activity, total anti-oxidant response (TAR), total peroxide (TPX), oxidative stress index (OSI) and some pro-oxidant cytokines in 29 children with CRF [mean age 10.2+/-3.5 years; 19 pre-dialysis, ten on continuous ambulatory peritoneal dialysis (CAPD)] and in 25 control subjects. Children with CRF had lower PON and TAR and higher TPX and OSI values than did controls (P<0.05). Except for lower TAR and serum albumin levels of the CAPD subgroup (P<0.05), other parameters were similar in non-dialysis and CAPD patients (P>0.05). Patients had significant positive correlation between TAR and serum albumin (P<0.05). Serum urea had significant positive correlation with TPX and OSI (P<0.05). Increased oxidative stress and decreased anti-oxidants measured by serum PON activity and TAR were found in children with CRF. We can hypothesize, on the basis of statistical correlations, that low levels of serum albumin and high levels of uraemic metabolites might be responsible for increased oxidative stress in children with CRF. Further studies with larger sample sizes are needed to verify these results. PMID- 16252098 TI - Renal transplant biopsy specimen adequacy in a paediatric population. AB - Updated guidelines on the diagnosis of acute allograft rejection including criteria for biopsy specimen adequacy were published in 1999. We sought to determine the adequacy of specimens in paediatric transplant patients and identify factors influencing adequacy. All renal transplant biopsies performed between 1998 and 2003 were classified as adequate (n =25), minimal (n =19) or inadequate (n =27) in accordance with the Banff 97 criteria, and the histological diagnoses were documented. The effect on specimen adequacy of grade of operator, method of sedation, age of child, needle gauge, number of cores and total core length was then investigated. Overall, a minimal or adequate specimen was obtained in 62% of cases. No histological diagnosis could be made in 30% of all specimens, just over half of which were inadequate. Higher rates of rejection were found in adequate (52%) than inadequate (33%) samples. The grade of operator (p =0.498), the age of the child at the time of biopsy (p =0.815) and type of sedation (p =0.188) did not affect adequacy. More than one core was obtained in 38 (54%) cases, and this was significantly associated with specimen adequacy (p <0.0005) as was longer total core length (p =0.002). Clinical features in isolation are not sufficient for the diagnosis of acute allograft rejection. Renal biopsy remains the gold standard and relies on adequate specimen collection. Our data shows that specimen adequacy according to the Banff 97 guidelines is achievable in children and that more than one core at the time of sampling significantly improves this achievement. Adequate sampling reduces the risk of an inconclusive histological diagnosis. PMID- 16252097 TI - Hypophosphatemic rickets: results of a long-term follow-up. AB - This study reports the benefits and side effects of conventional treatment, phosphate and calcitriol supplementation in patients with heritable hypophosphatemic rickets and a long-term follow-up, median of 60.9 months. The group is composed of 17 patients (ten girls). Sixteen patients presented with bone pain and/or deformities, and in one patient the diagnosis was radiological. All the patients had increased alkaline phosphatase, hypophosphatemia, decreased fractional phosphate tubular reabsorption (TRP) and maximum tubular phosphate reabsorption/glomerular filtration rate ratio (TPO4/GFR). Ten of 17 patients had metabolic acidosis, which was corrected only with the conventional treatment. Potassium citrate was prescribed to the patients who developed hypercalciuria. Excluding one patient with pulmonary dysfunction, the remaining 16 patients were divided into two groups according to the age at treatment onset (T0): group I (GI) > or =4 years (n =9) and GII <4 years (n =7). GI and GII had similar follow up periods and treatment protocols. Seven out of nine GI patients underwent orthopedic surgery, in contrast to none of GII. Anthropometric data results showed that within each group there is no difference in weight and stature z score at T0 and at the end of the observation (Tf), but, when both groups are compared, GII shows higher z-score for stature at T0 (p <0.05) and at Tf (p <0.05). Nephrocalcinosis developed in three cases and correlated with hypercalciuria (p <0.001) and dose of calcitriol (p =0.03). In conclusion, higher stature z-score is associated with early treatment. A careful protocol is recommended to detect such complications as nephrocalcinosis. We suggest potassium citrate for patients with hypercalciuria to avoid calcium precipitation. PMID- 16252095 TI - Report of an NIH task force on research priorities in chronic kidney disease in children. PMID- 16252099 TI - Tissue plasminogen activator for blocked peritoneal dialysis catheters. AB - Tissue plasminogen activator was used for a blocked peritoneal dialysis catheter in a child with no vascular access. The catheter was salvaged using tissue plasminogen activator and dialysis could be carried out without any difficulty. PMID- 16252101 TI - Long-term follow-up of extremely low birth weight infants. PMID- 16252100 TI - Childhood microscopic polyangiitis associated with MPO-ANCA. AB - We reviewed the clinical, histological and serological parameters of microscopic polyangiitis (MPA) associated with antineutrophil cytoplasmic antibodies (ANCA) specific to myeloperoxidase (MPO). Six girls and one boy aged 12.0+/-2.6 years (7 15 years) met the following inclusion criteria: (1) clinical manifestations of systemic small vessel involvement; (2) histological demonstration of pauci-immune necrotizing glomerulonephritis; and (3) serological findings of increased concentration of MPO-ANCA by ELISA test. The main clinical manifestations were: influenza-like symptoms (100%), hematuria/proteinuria (100%), purpura (100%), pulmonary-renal syndrome (57%), acute renal failure (ARF) (29%), ischemic cerebral insults (29%), and necrotizing vasculitis of the skin (29%). All patients underwent renal biopsy examined by immunohistochemistry with expression of alpha-smooth muscle actin (alpha SMA) in glomerular and interstitial spaces. Patients were followed from 6 months to 5.5 years (35.4+/- 23.2 months). None of the patients died. Two of seven children who had ARF progressed to end stage renal disease; one developed chronic renal failure, and four normalized renal function. ARF and central nervous system involvement at presentation were parameters of poor renal outcome. A high score of fibro-cellular glomerular crescents was associated with worse prognosis. Early treatment enables a favorable prognosis of MPO-ANCA-associated MPA in children. PMID- 16252102 TI - Recessive distal renal tubular acidosis in Sarawak caused by AE1 mutations. AB - Mutations of the AE1 (SLC4A1, Anion-Exchanger 1) gene that codes for band 3, the renal and red cell anion exchanger, are responsible for many cases of familial distal renal tubular acidosis (dRTA). In Southeast Asia this disease is usually recessive, caused either by homozygosity of a single AE1 mutation or by compound heterozygosity of two different AE1 mutations. We describe two unrelated boys in Sarawak with dRTA associated with compound heterozygosity of AE1 mutations. Both had Southeast Asian ovalocytosis (SAO), a morphological abnormality of red cells caused by a deletion of band 3 residues 400-408. In addition, one boy had a DNA sequence abnormality of band 3 residue (G701D), which has been reported from elsewhere in Southeast Asia. The other boy had the novel sequence abnormality of band 3 (Q759H) and profound hemolytic anemia. PMID- 16252104 TI - IgA deficiency and membranous glomerulonephritis. PMID- 16252105 TI - Clinical course of prenatally detected primary vesicoureteral reflux. AB - The purpose of this study was to report the clinical course of medium-long-term follow-up of children with prenatally detected vesicoureteral reflux (VUR). Between 1986 and 2004, 53 (41 males) children with VUR detected by investigation of prenatal hydronephrosis were followed up for a mean time of 66 months (range: 6-200 months). Newborns were investigated by ultrasound, voiding cystourethrogram (VCUG) and DMSA scan. Follow-up clinical visits were performed at 6-month intervals. After 24 months patients were investigated by conventional VCUG or direct isotope cystogram. Survival analysis was performed in order to evaluate the resolution of the reflux. Differences between subgroups (mild vs moderate/severe reflux) were assessed by the two-sided log rank test. Thirty (58%) infants presented bilateral VUR, for a total of 83 renal units. There was a predominance of severe reflux (54%). Renal damage was detected in 33.7% of the units on first renal scan. There was a significant correlation between severe reflux and renal damage scars (RR=3.4, 95% confidence interval [CI], 1.4-8, p=0.002). Forty-seven patients were treated with continuous prophylaxis. One patient developed systolic hypertension. Urinary tract infection occurred in 12 (25%) children conservatively managed. VUR resolution was evaluated in 56 renal units. Spontaneous resolution was observed in 25 units (45%). At 48 months after diagnosis, 75% of the cases of mild reflux (I-III) and 37% of severe reflux (IV V) had resolved (log-rank, 5.6, p=0.017). There was an improvement of nutritional parameters between admission and the end of follow-up. In conclusion, the clinical course of prenatally detected VUR followed up on a medium-long-term basis is relatively benign. Our study corroborates the results obtained in other series of infants with reflux that emphasized the heterogeneity of this disorder. PMID- 16252106 TI - Potential cardiovascular risk factors in paediatric renal transplant recipients. AB - Cyclosporin (CsA) therapy is associated with side effects such as hypertension, hyperlipidemia and nephrotoxicity. Tacrolimus (Tac) has been shown to be more favourable in this respect. We retrospectively analysed office blood pressure (BP), serum total cholesterol (TC) and fasting glucose levels, and estimated graft function profiles in paediatric (n =56) and young adult (n =14) renal transplant recipients whose maintenance immunosuppressive regimen was based upon CsA (n =38) or Tac (n =32) given with mycophenolate mofetil and corticosteroids. The analysis was performed at four different time-points: at 1, 6, 12, and 24 months post-transplant, respectively. Baseline characteristics were comparable between treatment groups. Differences for both systolic and diastolic BP, and graft function between treatment groups became significant from month 1 and throughout the 2-year period. Values (mean +/- SD) for CsA-treated and Tac treated recipients at 2 years were 118.8+/-11.1 / 74.6+/-7.4 mmHg vs 109.3+/-11.2 / 67.2+/-7.8 mmHg for systolic and diastolic BP, respectively, p <0.005/0.005; and 72.0+/-18.5 ml/min vs 84.0+/-22.4 ml/min per 1.73 m(2) for graft function, respectively, p <0.01. Office hypertension, defined as the use of antihypertensive medication at month 24, was significantly associated with CsA therapy (chi(2), p <0.01). TC levels became significantly lower at months 6, 12, and 24 in the Tac group compared with the CsA group. Hypercholesterolemia, defined as TC>or=200 mg/dl, was significantly associated with CsA-based immunosuppressive regimen at months 6, 12, and 24 post-transplant (chi(2), p <0.05, p <0.001, and p <0.01, respectively). Although Tac therapy was associated with higher glucose levels, no recipient developed post-transplant diabetes mellitus. The number of recipients who experienced acute rejections was comparable in both groups. In conclusion, Tac-based immunosuppressive therapy was found to be associated with more favourable potential risk-factor profiles for cardiovascular disease and better graft function at 2 years post-transplant compared with CsA-therapy. PMID- 16252107 TI - Strict cysteamine dose regimen is required to prevent nocturnal cystine accumulation in cystinosis. AB - Cystinosis is an autosomal recessive disorder, caused by mutations in the lysosomal cystine carrier cystinosin, encoded by the CTNS gene. The disease generally manifests with Fanconi syndrome during the first year of life and progresses towards end stage renal disease before the age of 10 years. Cysteamine depletes intralysosomal cystine content, postpones the deterioration of renal function and the occurrence of extra-renal organ damage. Based on the pharmacokinetic data, patients with cystinosis are advised to use cysteamine every 6 h. The aim of this study was (1) to evaluate the cysteamine dose regimen in Dutch patients with cystinosis and (2) to determine morning polymorphonuclear (PMN) leukocyte cystine content 6 h vs 9 h after the last evening cysteamine dose. Only 5/22 of Dutch cystinosis patients ingested cysteamine every 6 h. Morning (8 a.m.) PMN cystine content in 11 examined patients was elevated 9 h after 12.5-15 mg/kg evening cysteamine dose compared to the value 6 h after the ingestion of the same dose (0.73+/-0.81 nmol vs 0.44+/-0.52 nmol cystine/mg protein, p =0.02). In conclusion, only the minority of Dutch cystinosis patients follows the recommended strict cysteamine dose regimen. We provide evidence that cysteamine has to be administered every 6 h, including the night, as it has much better effect for maintaining low PMN cystine levels. PMID- 16252108 TI - The efficiency of intraperitoneal high-dose immunoglobulin in experimental nephrotic syndrome. AB - Although it has been reported that high-dose immunoglobulin has beneficial effects in chronic glomerulonephritis, it is not known whether it is effective in the treatment of idiopathic nephrotic syndrome. We have investigated the effects of intraperitoneal immunoglobulin in adriamycin-induced nephrotic syndrome. Adriamycin (2 mg kg(-1) per dose) was given intravenously to sixteen Wistar albino rats (eight control and eight treatment rats) on day 1 and at week 3. At week 5 intraperitoneal immunoglobulin (1 g kg(-1) per dose) was given to the treatment group on two consecutive days whereas the control group received intraperitoneal saline solution. In both treatment and control groups urinary protein excretion was significantly elevated after administration of adriamycin (P=0.018). Urinary protein excretion, serum albumin, and triglyceride levels in the two groups were not significantly different after 5, 8, 12, and 16 weeks. Serum creatinine levels were higher and creatinine clearance was significantly lower in the control group in week 16 (P=0.001 and P=0.049, respectively). Glomerular sclerosis index was significantly lower in the treatment group (P=0.012). Although intraperitoneal high-dose immunoglobulin did not reverse biochemical results, it is encouraging that glomerular sclerosis index was significantly lower in the treatment group. PMID- 16252109 TI - Hypokalemia causing rhabdomyolysis resulting in life-threatening hyperkalemia. AB - Hyperkalemia is commonly associated with renal failure and is rare if renal function is normal. It is rarely caused by rhabdomyolysis and can be life threatening if unrecognized. Rhabdomyolysis is caused by many disorders, including hypokalemia. The available literature, both for human beings and experimental animals, contains evidence of hypokalemia causing rhabdomyolysis and rhabdomyolysis leading to hyperkalemia. Hypokalemia as a cause of rhabdomyolysis often goes unnoticed, because of the counteracting effect of rhabdomyolysis on serum potassium. There are no pediatric reports showing hypokalemia as a cause of rhabdomyolysis leading to life-threatening hyperkalemia. This case emphasizes the vigilance required toward the occurrence of rhabdomyolysis, which if not detected promptly may prove fatal. PMID- 16252110 TI - Stent-related ureteric obstruction in paediatric renal transplantation. AB - The rates of ureteric obstruction and complications for use of externally draining uretero-vesico-cutaneous (external) stents (Group 1: n=39) and the use of internal uretero-vesical (double-J) stents (Group 2: n=16), in 55 of 64 consecutive paediatric renal-transplant recipients, performed at our institution between January 1996 and December 2003, have been compared. Serum creatinine levels pre and post-operatively and pre and post-stent removal were recorded. The diagnosis of ureteric obstruction was based on an increase in serum creatinine of >or=20%, in conjunction with ultrasound evidence of hydronephrosis or hydroureter, where other causes of renal dysfunction were excluded. Ureteric obstruction occurred in 13 of the 39 patients (33.3%) in Group 1, compared with only one case of ureteric obstruction in the 16 patients (6.25%) in Group 2 (OR=7.5, 95% CI=0.8-70, P=0.038). There was no evidence of a difference in the number of urinary tract infections (9/39 in Group 1, 6/16 in Group 2, OR=0.5, 95% CI=0.14 to 1.8, P=0.275) or the mean length of hospital stay (10.9 days in Group 1, 10.1 days in Group 2, 95% CI=-2.3 to 4 days, P=0.565) between the two groups. Glomerular filtration rate (GFR) improved in the week after stent removal in Group 2, but deteriorated in Group 1 (P=0.07). This non-randomised comparison of stent types supports the use of prophylactic double-J stents in paediatric renal transplantation- in terms of decreased ureteric complications and improved renal function post-stent removal. PMID- 16252111 TI - Factors associated with mortality in acute renal failure (ARF) in children. AB - OBJECTIVE: To assess the factors that affect the mortality in acute renal failure (ARF) in children. PATIENTS AND METHODS: We studied 149 patients with ARF and described the findings by age, gender, pathophysiological mechanism of renal damage, and type of renal damage, which can be oligoanuric and/or septic. We used multiple logistic analysis, Cox analysis for survival, and Kaplan-Meier curves. RESULTS: The male/female ratio was 91/58. The most affected age groups were newborns (44.3%) and infants (37.6%). The ARF mechanism was ischemic in 87 cases (58%) and the most frequent clinical type was nonoliguric in 118 cases (79.2%). In the multiple logistic regression analysis, only oliguria (P=0.07) and age group (P=0.049) were associated with mortality. In the survival analysis using the Cox method, oliguria (P=0.003) and sepsis (P=0.03) were associated with mortality. The survival curves showed that the cumulative probability of dying in the first 10, 20, or 40 days after the event was 75, 70, and 45% respectively. When oliguria was present, the survival at day 10 was 47% and when sepsis was present it was 68%. CONCLUSION: Oliguria, age, and sepsis are factors associated with mortality in children with ARF. PMID- 16252112 TI - Hypocitraturia: a risk factor for reduced bone mineral density in idiopathic hypercalciuria? AB - The association between idiopathic hypercalciuria (IH) and reduced bone mineral density (BMD) has been described in adults and children. Frequently, hypocitraturia (HC) is an associated condition. To determine the effect that HC may have on bone metabolism of these patients, we studied 88 children with IH at diagnosis, divided into the following groups: group 1-44 (50%) patients with associated HC; group 2-44 (50%) patients without HC; group 3 (29 subjects), a healthy control group. Urinary and blood electrolytes, as long as urinary N telopeptide, were measured. Lumbar spine (L2-L4) and femoral neck bone mineral density (BMD) and bone mineral content (BMC) were measured by dual energy X-ray absorptiometry. There was no difference in age between the three groups (P=0.80), but weight, height, body mass index, and bone age were lower (P<0.01) and serum intact parathyroid hormone (iPTH) was higher (P<0.05) in group 1 than in groups 2 and 3. N-telopeptide, measured in urine, did not differ between groups. The following bone densitometry parameters: lumbar spine BMC, BMC adjusted for height (BMCh), BMC adjusted for width of vertebrae (BMCw) and BMD, as well as femoral neck BMD, were significantly lower in group 1 than in groups 2 and 3 (P<0.01). When we corrected densitometry parameters for height, BMC was lower in group 1 and not in group 2 when compared with controls. CONCLUSIONS: Children with IH and associated HC may have a higher risk of bone mass loss and consequent osteopenia. Further studies are needed to assess the role that hypocitraturia may have in this form of bone disease. PMID- 16252113 TI - Unilateral renal angiodysplasia in a girl with hypertension. AB - We report a case of very unusual renal vascular anomaly: angiodysplasia. The patient suffered from acute pyelonephritis immediately after birth. Renal ultrasonography, performed at age 1 day, revealed an enlarged left kidney with heterogeneously increased echogenicity, which involuted rapidly in 3 months. At age 10 years, she presented with severe hypertension during a course of acute pyelonephritis. Peripheral plasma renin activity was high. Computed tomographic angiogram revealed a very small but functioning left kidney. A single, narrow, left renal artery did not have focal stenosis. Pathology examination revealed dysplastic arterioles at the subcapsular area. After left nephrectomy, the blood pressure and the plasma renin activity were normalized. In conclusion, this is an unusual case of renal angiodysplasia, which induced renin-dependent hypertension. PMID- 16252116 TI - [Interface pressure measurement in the treatment of pressure sores. Comparison with subcutaneous pressure values measured in healthy volunteers]. AB - BACKGROUND: Pressure relief is an important factor to determine the quality of anti-decubitus devices. Many studies have been carried out to measure the interface pressure, but because the results differed markedly, we made a study to assess the measurement devices. METHODS: The systems FSA, a device for internal use in the Luck company, the X-Sensor and an invasive subcutaneous pressure measurement over the os-sacrum (COACH: piezoelectric probe, MIPM) were tested on 20 healthy volunteers (average age 27 years, average BMI 21.8). RESULTS: The values of interface measurement showed obvious deviations from subcutaneous pressure in two cases (FSA +1.8%, Luck -33%, X-Sensor +65%) and some results showed extreme variations (FSA +18 mmHg, Luck -21 mmHg, X-Sensor +27 mmHg). CONCLUSIONS: The interface pressure measurement (with a few volunteers) is not suitable for a correct assessment of anti-decubitus devices. Furthermore, the use of this method is not necessary to monitor pressure relief in intensive care patients. These systems can, however be useful in the training of nursing staff. PMID- 16252114 TI - [Management of the upper airway in spontaneously breathing children. A challenge for the anaesthetist]. AB - In unconscious, spontaneously breathing and anaesthetised children, a high incidence of partial or complete airway obstruction jeopardizes sufficient oxygenation. In this situation, the most important and efficient manoeuvre is to open up the upper airway. Chin lift, jaw thrust and continuous positive airway pressure (CPAP) are proven and effective methods for opening an obstructed upper airway. In addition to these simple airway manoeuvres, different techniques of body positioning (e.g., lateral positioning or supine position in combination with the "sniffing position") are effective to improve and maintain upper airway patency. PMID- 16252118 TI - Comparison of seedling recruitment under arborescent palms in two Neotropical forests. AB - Certain overlying strata in forests may disproportionately reduce seedling density and species richness. For eight arborescent palm species, we quantified the relative restriction of seedling recruitment under individual palms versus non-palm sites and extended to the landscape scale by quantifying the total area covered by arborescent palms at Barro Colorado Island (BCI), Panama and La Selva Biological Station, Costa Rica. We also examined whether differences among palm species in restricting seedling recruitment were associated with differences in crown architecture, litter depth, and light availability. Woody seedlings had lower mean density/m2 and mean number of species/m2 under individual palms than at non-palm sites for all four palm species at BCI, but for none at La Selva. Estimated species richness for woody seedlings, derived via rarefaction, was lower under palm than non-palm microsites at both BCI and La Selva, but not for non-woody seedlings. Differences in seedling density corresponded to some key architectural characters that differed among the palm species. Light availability was lower under palm than non-palm microsites at both BCI and La Selva, but only estimated species richness of woody seedlings at BCI was strongly correlated with % canopy openness. The coverage of arborescent palms was much lower at BCI than La Selva. Therefore, at BCI, the relative restriction of woody seedling recruitment under individual palms does not accumulate greatly at the landscape scale. At La Selva, for woody seedlings, only estimated species richness was relatively limited under palms, and non-woody seedlings had relatively lower mean density/m2 and mean number of species/m2 under only one palm species. Therefore, the relative restriction of seedling recruitment by arborescent palms at La Selva is limited at both individual and landscape scales. PMID- 16252119 TI - Interactions between a detrital resource pulse and a detritivore community. AB - Detritivore communities influence the decomposition of detrital resources in virtually all natural systems. Conversely, detrital resources can also have considerable bottom-up effects on detritivore communities. While many investigations have examined detritivory and decomposition processes, few have considered interactions between detritivores and detritus as concurrent processes in the same system, or in the context of natural detrital pulses. In many systems, resource pulses contribute substantial detrital inputs to belowground systems. These detrital pulses may influence interactions between the detritivore community and detrital decomposition. I conducted field experiments to investigate interactions between detrital resource pulses of periodical cicada (Magicicada spp.) carcasses and scavenging detritivorous macroarthropods. Cicada litterfall pulses influenced several broad groups in the macroarthropod community, including relatively specialized necrophilous taxa and relatively generalized detritivores, omnivores and predators. Conversely, detritivore activity increased the rate of cicada carcass decomposition by 4,082% compared to caged control carcasses. These results suggest that interactions between pulses of cicada detritus and the detritivore community influence both the persistence of ephemeral detrital resources, and the distribution, abundance and behavior of detritivore populations. PMID- 16252120 TI - Cataloging transcription factor and major signaling molecule genes for functional genomic studies in Ciona intestinalis. AB - The ascidian Ciona intestinalis provides an excellent experimental system for functional genomic studies because (1) its genome has been sequenced, (2) the transcription factor genes and genes for major signal transduction molecules have been extensively screened and annotated on a genome-wide scale using the molecular phylogenetical method, and (3) their embryonic expression profiles have been almost completely determined. However, the entire genetic structure, including the 5' and 3' untranslated regions and the protein-coding regions, of most gene models used in these prior studies is not always supported by cDNA evidence, and thus, these gene models are potentially imprecise. To facilitate functional genomic studies based on precise gene structures, our present study determined 406 cDNA sequences for 357 transcription factor genes and 112 cDNA sequences for 107 signal transduction molecule genes, greatly improving the previous gene models and revealing transcript variants for 44 genes. Considering these data alongside those of previously characterized genes deposited in the DNA Data Bank of Japan/European Molecular Biology Laboratory/GENBANK databases, 95.6% of the catalogued transcription factor genes (373/390) and 98.3% of the catalogued signal transduction molecule genes (117/119) have now been verified by cDNA sequences. Thus, the present study greatly improves the resources available for functional genomic studies in C. intestinalis. PMID- 16252122 TI - An autologous system for culturing head and neck squamous cell carcinomas for the assessment of cellular therapies on the chorioallantois membrane. AB - The evaluation of novel immunotherapeutic anti-cancer strategies requires target and effector cells from the autologous host to avoid false positive results because of allo-reactivity. Head and neck squamous cell carcinoma cells (HNSCC) do not proliferate under standard culture conditions in plastic ware. To create an autologous model, the chorioallantois membrane (CAM) of chicken embryos was employed to culture HNSCC and peripheral blood mononuclear cells (PBMC). Separated tumor cells were co-incubated with a cytostatic agent, PBMC, or mere cell culture medium as a control. Tumor cell lysis was assessed by acridine orange staining or by flow cytometry analysis of iodide marked cells after 24 and 48 h of co-incubation. Incubation with cisplatin resulted in a decrease of viable cells by 49% after 24 h and 48 h. In contrast, incubation with mere culture medium resulted in an increase of viable tumor cells by 5% after 24 h and a decrease of 4% after 48 h; the incubation of tumor cells and PBMC led to a non significant decrease by 14% after 24 h and 16% after 48 h. Taken together, the CAM supports a favorable environment for the co-culture of HNSCC and PBMC, thus providing an approximated in vivo autologous model to assess the efficacy of new cell-therapeutical approaches. PMID- 16252121 TI - Degenerate coding in neural systems. AB - When the dimensionality of a neural circuit is substantially larger than the dimensionality of the variable it encodes, many different degenerate network states can produce the same output. In this review I will discuss three different neural systems that are linked by this theme. The pyloric network of the lobster, the song control system of the zebra finch, and the odor encoding system of the locust, while different in design, all contain degeneracies between their internal parameters and the outputs they encode. Indeed, although the dynamics of song generation and odor identification are quite different, computationally, odor recognition can be thought of as running the song generation circuitry backwards. In both of these systems, degeneracy plays a vital role in mapping a sparse neural representation devoid of correlations onto external stimuli (odors or song structure) that are strongly correlated. I argue that degeneracy between input and output states is an inherent feature of many neural systems, which can be exploited as a fault-tolerant method of reliably learning, generating, and discriminating closely related patterns. PMID- 16252123 TI - Bilateral and unilateral internal carotid artery dissection causing isolated hypoglossal nerve palsy: a case report and review of the literature. AB - Two cases of isolated hypoglossal nerve palsy due to internal carotid artery dissection are reported. In one case, a bilateral dissection was encountered. The clinical presentation and multidisciplinary management are discussed and the literature reviewed. PMID- 16252124 TI - Is laryngopharyngeal reflux an important risk factor in the development of laryngeal carcinoma? AB - Laryngopharyngeal reflux (LPR) as a causative irritating factor in the development of laryngeal carcinoma has been suggested. However, the association between LPR and laryngeal carcinoma remains unclear. The aim of this study was to compare the prevalence and severity of reflux between patients with laryngeal carcinoma and clinical LPR. The intention was to find out if a correlation exists between the laryngeal findings and the level of acid reflux. The study population consisted of 29 patients with laryngeal cancer (group I), 33 LPR patients with normal laryngeal findings (group II) and 70 LPR patients with related laryngeal pathology (group III). The results of 24-h, double-channel ambulatory esophageal pH monitoring were analyzed comparing the three groups. The occurrence and severity of abnormal acid reflux at the upper and lower esophageal segments were evaluated. The incidence of LPR or gastroesophageal reflux (GER) did not vary in any of the three groups (LPR was present in 62, 42 and 56% of the patients, while GER was present in 45, 24 and 37% of the patients, respectively). Patients with LPR or GER from the three groups did not differ significantly in terms of the number of acid reflux episodes and percentage of times when the pH was <4. Our data do not support the hypothesis that LPR can be an independent risk factor in the development of larynx cancers. However, the data also do not thoroughly exclude the possibility. The reason why LPR leads to variable pathologies in the larynx may be uncovered by studies probing the differences between patients via detailed examinations of the local anti-reflux barriers such as epithelial morphology and functions. PMID- 16252125 TI - Reconstruction of massive rotator cuff lesions with a synthetic interposition graft: a prospective study of 41 patients. AB - In the present prospective study, we report about our experience with massive rotator cuff tears treated by means of a non-resorbable transosseously fixated patch combined with subacromial decompression. Forty-one patients were followed clinically and radiographically for a mean duration of 43 months. Their mean preoperative Constant and Murley score improved (P<0.001) from 25.7 preoperatively to 72.1 postoperatively. Substantial pain relief and improvement in the performance of activities of daily living were obtained. Anatomically, the repair resulted in a mean acromiohumeral interval of 8.6 mm. At the latest follow up, three patients presented with a new tear between the inserted mesh and the supraspinatus musculotendinous unit. Reoperations were not performed. For short term periods, restoring a massive rotator cuff tendon defect with synthetic grafts combined with subacromial decompression can give significant pain relief and improvement of range of motion and strength with few complications. PMID- 16252126 TI - [MRI after meniscus and cruciate ligament surgery]. AB - Nowadays, due to its high diagnostic accuracy, MR imaging is the method of choice for the evaluation of knee joint disorders pre- and postoperatively. Accurate diagnosis is sometimes possible only if the reporting radiologist has knowledge of the therapeutic procedures and the surgical report. Frequently, further therapeutic management is strongly influenced by MR examinations and radiological reports. PMID- 16252127 TI - [Improvements in detection of rectal cancer recurrence by multiplanar reconstruction]. AB - PURPOSE: To evaluate the advantages of multiplanar reconstruction and different axial slice thickness in diagnostic of rectal cancer recurrence after operation and radiotherapy. METHOD: We included 83 patients after operation and radiotherapy of rectal cancer in this study. All patients got a minimum of three CT-examinations in their follow-up program. A total of 294 CT-scans were evaluated. Each examination was reviewed by two experienced radiologists in respect to recurrence. Each examination was presented in axial reconstruction with a slice sickness of 8, 5, and 1.25 mm and in multiplanar reconstruction. The sensitivity, specificity, positive predictive value and accuracy were calculated. RESULTS: Multiplanar reconstructions showed better results for the detection of recurrence than axial reconstruction. A reduced slice thickness did not lead to better results in axial reconstruction. Multiplanar reconstruction showed a sensitivity of 0.88, a specificity of 0.98, an accuracy of 0.96 and a positive predictive value of 0,94, for axial reconstruction we calculated: 0.82, 0.97, 0.94 and 0.88, respectively. Sensitivity and accuracy showed a significant increase after the first and second examination. CONCLUSION: Multiplanar reconstructions allow a significant better detection of rectal cancer recurrence when compared to axial reconstructions. Thinner axial slice thickness shows no diagnostic advantage. PMID- 16252128 TI - Comparative chemical characterization of pigmented and less pigmented cell walls of Alternaria tenuissima. AB - Alternaria tenuissima, the parasitic fungus, was obtained from the pruned upper cut surfaces of mulberry stems. This fungus contains dark pigment because of the presence of melanin in the cell wall. To obtain less-pigmented cell walls, this fungus was grown under dark condition. When the pigmented and less-pigmented cell walls were chemically analyzed, no differences were observed in amino-acid composition, hexoses, or pentoses. However, in pigmented cell walls, higher contents of melanin (2.6%) were found than in less-pigmented cell walls (0.3%). Interestingly, a significant difference was observed in the relative fatty-acid compositions between these two types of cell walls. Among the major fatty acids, there were increased concentrations of tetradecanoic acid (C14:0), hexadecanoic acid (C16:0), 9-hexadecenoic acid (C16: 1,Delta 9), and 9-octadecanoic acid (C18:1,Delta 9) and a concomitant decrease in 9,12-octadecadienoic acid (C18:2,Delta 9,12) in less-pigmented compared with pigmented cell walls. This difference in fatty-acid composition may be related to the higher percentage of melanin in the pigmented than the less-pigmented cell walls. Lesser amounts of 9,12-octadecadienoic acid in less-pigmented cell walls may have been caused by the growth of the fungus under environmental stress conditions. An interesting observation was the presence in pigmented cell walls only of methyl-substituted fatty acids with carbon numbers C14 to C17, but their occurrence could not be ascertained in the present study. PMID- 16252129 TI - Discovery of a novel Wolbachia super group in Isoptera. AB - Wolbachia are one of the most abundant groups of bacterial endosymbionts in the biosphere. Interest in these heritable microbes has expanded with the discovery of wider genetic diversity in under-sampled host species. Here, we report on the putative discovery of a new genetic lineage, denoted super group H, which infects the Isopteran species Zootermopsis angusticollis and Z. nevadensis. Evidence for this novel super group is based on portions of new Wolbachia gene sequences from each species spanning 3.5 kilobases of DNA and the following genes: 16S rDNA, dnaA, gltA, groEL, and ftsZ. Single-gene and concatenated maximum likelihood phylogenies establish this new super group and validate the positioning of the other Wolbachia super groups. This discovery is the first example of a termite Wolbachia that is highly divergent from the Isopteran Wolbachia previously described in super group F. This study highlights the importance of multilocus approaches to resolving Wolbachia super group relationships. It also suggests that surveys of Wolbachia in more earlier-originating (and under-sampled) groups of arthropods are more apt to reveal novel genetic diversity. PMID- 16252130 TI - Characterization of exopolysaccharides produced by cyanobacteria isolated from Polynesian microbial mats. AB - Six cyanobacterial isolates recovered from Polynesian microbial mats, called "kopara," were cultured using laboratory-closed photobioreactors and were shown to produce exopolymers as released and capsular exopolysaccharides (EPS). These polymers have been chemically characterized using colorimetric and elemental assays, infrared spectrometry, and gas chromatography. Both capsular and released EPS consisted of 7 to 10 different monosaccharides with neutral sugars predominating. Interestingly, four isolates exhibited sulfate contents ranging from 6% to 19%. On the basis of preliminary data, cyanobacteria from this unusual ecosystem appear to be an important source of novel EPS of a great interest in terms of their biological activities. PMID- 16252131 TI - The major Aeromonas veronii outer membrane protein: gene cloning and sequence analysis. AB - The gene encoding the major outer membrane protein (OMP) from Aeromonas veronii, Omp38, was cloned and characterized. Sequence analysis revealed an open reading frame of 1,047 nucleotides coding for a primary protein of 349 amino acids with a 20-amino-acid signal peptide at the N-terminal and the consensus sequence Ala-X Ala (Ala-Asn-Ala) as the signal peptidase I recognition site. The mature protein is composed of 329 amino acids with a calculated molecular mass of 36,327 Da. The degree of identity of the deduced Omp38 amino acid sequence to porins from enteric bacteria (OmpF, PhoE, and OmpC) was only 30%. Nevertheless, Omp38 possesses typical features of Gram-negative porins, including acidic pI, high glycine and low proline content, no cysteine residues, and a carboxy-terminal Phe. On the basis of PhoE-OmpF three-dimensional structure and the Kyte-Doolittle hydrophobicity analysis, it seems likely that Omp38 secondary structure consists of 16 antiparallel beta-strands and 8 loops. Phylogenetic analyses among Omp38 and related porins from Gram-negative bacteria originate well-defined clusters that agree with the taxonomy of the corresponding organisms. PMID- 16252132 TI - Purification and characterization of Cry1Ac toxin binding proteins from the brush border membrane of Helicoverpa armigera midgut. AB - Several Cry1Ac binding proteins from midgut of Helicoverpa armigera were purified using toxin-affinity chromatography. Enzyme assays showed that the purified proteins had strong aminopeptidase activity. The N-terminal sequences confidently identified a 124-kDa binding protein as an aminopeptidase N (APN), and some similarity suggests that a 162-kDa binding protein may also be an APN. Two minor binding proteins were not characterized. PMID- 16252134 TI - Expression of the Bacillus thuringiensis mosquitocidal toxin Cry11Aa in the aquatic bacterium Asticcacaulis excentricus. AB - A mosquitocidal aquatic bacterium has been developed by introducing an operon containing the cry11Aa, and p20 genes from Bacillus thuringiensis subsp. israelensis (Bti) into the gram-negative aquatic bacterium Asticcacaulis excentricus. After transformation, the cry11Aa gene was successfully expressed in recombinant A. excentricus under the tac promoter, at the level of 0.04 pg/cell. The recombinant bacteria were toxic to Aedes aegypti larvae with an LC(50) of 6.83 x 10(5) cells/mL. We believe that these bacteria may have potential as genetically engineered microorganisms for the control of mosquito larvae. PMID- 16252133 TI - A comparison of a new oral streptogramin XRP 2868 with quinupristin-dalfopristin against antibiotic-resistant strains of haemophilus influenzae, Staphylococcus aureus, and Streptococcus pneumoniae. AB - A new streptogramin antibiotic XRP 2868 was compared with quinupristin dalfopristin for inhibitory activities against antibiotic-resistant Haemophilus influenzae, Staphylococcus aureus, and Streptococcus pneumoniae. In each organism examined, XRP 2868 had an IC(50) that was twofold to fivefold lower than quinupristin-dalfopristin, for inhibition of cell viability, protein synthesis, and ribosomal subunit formation. PMID- 16252135 TI - Docetaxel/Gemcitabine followed by gemcitabine and external beam radiotherapy in patients with pancreatic adenocarcinoma. AB - BACKGROUND: Pancreatic cancer remains highly lethal. Previous attempts with neoadjuvant therapy in this disease have been inconclusive, but a potential for benefit exists. We conducted a phase II trial of dose-intense docetaxel and gemcitabine followed by twice-weekly gemcitabine and external beam radiotherapy in patients with pancreatic adenocarcinoma. METHODS: Patients with stage I to III disease were eligible. Docetaxel 65 mg/m(2) intravenously over 1 hour and gemcitabine 4000 mg/m(2) given intravenously over 30 minutes were given on days 1, 15, and 29. On day 43, radiotherapy was begun at 50.4 Gy with gemcitabine 50 mg/m(2) intravenously over 30 minutes twice weekly for 12 doses. After treatment, patients were considered for resection. RESULTS: Twenty-four assessable patients were recruited onto the trial. All but one patient completed a full 12 weeks of therapy. Grade 3 and 4 hematological and nonhematological toxicities were common but manageable, and neutropenic fever did not occur. No patient had local tumor progression. Twelve patients (50%) responded by Response Evaluation Criteria in Solid Tumors Group (RECIST) criteria, including one radiographic complete response. Seventeen patients underwent resection after therapy. Margin-negative resections were performed in 13 patients, including 9 patients whose disease was borderline or unresectable before treatment. A treatment effect was seen in all resection specimens. There have been no local recurrences of tumor, and several patients remain alive without evidence of disease. CONCLUSIONS: Docetaxel/gemcitabine followed by gemcitabine/radiotherapy is active in the treatment of pancreatic adenocarcinoma, with manageable toxicity. Tumor downstaging occurs in some patients to allow complete resection. Further investigation of this regimen is warranted. PMID- 16252136 TI - The surgical and functional outcome of limb-salvage surgery with vascular reconstruction for soft tissue sarcoma of the extremity. AB - BACKGROUND: This study compared the surgical, oncological, and functional outcomes of patients undergoing limb-salvage surgery for extremity soft tissue sarcoma with vascular resection and reconstruction with the outcomes of those undergoing limb-salvage without vascular reconstruction. METHODS: Nineteen patients were identified from a prospective soft-tissue sarcoma database who underwent vascular resection and reconstruction as part of their limb-salvage surgery and who were followed up for at least 1 year or until death. Each of these 19 patients was case-matched to 2 additional patients on the basis of tumor location, size, and depth; patient age; and timing of radiation. To compare functional outcome, a subset of patients was case-matched with additional criteria including wound-complication status, motor nerve sacrifice, similar preoperative function as determined by the Toronto Extremity Salvage Score, and no metastases at diagnosis or the 1-year follow-up. RESULTS: Patients in the vascular reconstruction group were more likely to require a muscle transfer (53% vs. 18%; P = .008), experience a wound complication (68% vs. 32%; P = .03), experience deep venous thrombosis (26% vs. 0; P = .003), experience significant limb edema (87% vs. 20%; P = .001), and ultimately require an amputation (16% vs. 3%; P = .07). Patients who underwent vascular reconstruction had only slightly lower Toronto Extremity Salvage Score scores 1 year after surgery (78.5 vs. 84.2; P = .35). There were no significant differences in local or systemic tumor relapse between the two groups. CONCLUSIONS: Vascular reconstruction is a feasible option in limb-salvage surgery for soft tissue sarcoma but is associated with an increased risk for postoperative complications, including amputation. Although function is not significantly worse after vascular reconstruction, the results are less predictable. PMID- 16252137 TI - Malignant vascular tumors: clinical presentation, surgical therapy, and long-term prognosis. AB - BACKGROUND: The aim of this study was to analyze the presentation of, surgery for, and prognosis of malignant vascular tumors (MVTs). METHODS: This was an observational single-center study. Patients who underwent operation for MVTs between 1988 and 2004 were included. Data were gathered prospectively in a computerized registry. RESULTS: Of 568 adult patients with soft tissue malignancies, 43 (7.6%) were treated for MVTs. Twenty-four men and 19 women (median age, 55.3 years) were referred for 30 primary tumors and 13 recurrences. Symptoms were observed in 90.7% of the cases (swelling [37.2%], pain [34.9%], extrusion [11.6%], hemorrhage [7%], weight loss [4.7%], loss of energy [4.7%], impaired function [4.7%], and others [30.2%]). Tumors were located in the extremities (n = 16), trunk (n = 3), abdomen (n = 15), retroperitoneum (n = 7), and thyroid gland (n = 2). Twenty-two (51.2%) angiosarcomas, nine (20.9%) malignant hemangiopericytomas, eight (18.6%) malignant epithelioid hemangioendotheliomas, and four (9.3%) lymphangiosarcomas were seen. The median overall survival after surgery was 21.4 months, with 2-, 5-, and 10-year overall survival rates of 41.5%, 38.3%, and 18.8%, respectively. MVTs of the extremities and trunk and localized disease indicated a better prognosis than abdominal or retroperitoneal MVTs (univariate and multivariate analyses: P = .0122 and P = .0287) and metastasized stages (univariate and multivariate analyses: P = .0187 and P = .0287). CONCLUSIONS: A considerable number of patients with soft tissue malignancies undergo surgery for MVT. Various symptoms and a multilocular occurrence are typical. The course of MVTs is aggressive. Tumor site and stage are important prognostic factors. Surgery is potentially curative, especially for localized disease of the extremities and trunk. PMID- 16252138 TI - Improved survival associated with postoperative wound infection in dogs treated with limb-salvage surgery for osteosarcoma. AB - BACKGROUND: Limb-salvage surgery and adjuvant chemotherapy are performed as a treatment of appendicular osteosarcoma in dogs. Approximately 50% of dogs that undergo limb-salvage surgery develop postoperative surgical wound infections. Postoperative surgical infections may affect survival in cancer patients. The purposes of this study were to examine the effect of surgical wound infection on survival, local recurrence, and metastasis in relation to other prognostic factors for dogs with spontaneous osteosarcoma treated with limb-salvage surgery. METHODS: Forty-seven client-owned dogs with osteosarcoma of the distal radius were treated with limb-salvage surgery and adjuvant chemotherapy--either carboplatin or carboplatin and doxorubicin. Hazard ratios were estimated by using the Cox proportional hazard model, and survival functions were estimated by using the Kaplan-Meier product-limit life-table method. RESULTS: Of the 47 dogs in this study, 32 (68%) developed a postoperative wound infection. Infection, dog weight, and extent of the primary tumor (percentage of length) significantly affected survival, and infection and percentage of length significantly affected time to metastasis. None of the variables considered in this study affected local recurrence. Dogs that were diagnosed with an infection were less likely to die (hazard ratio, .446), and dogs with greater body weight and greater percentage length involvement were more likely to die (hazard ratios of 3.37 and 3.66, respectively). CONCLUSIONS: In dogs with osteosarcoma treated with limb-salvage surgery, infection has a positive influence on survival, as does a smaller initial length of radius involved and lower body weight. PMID- 16252139 TI - Low-attenuation oral GI contrast agents in abdominal-pelvic computed tomography. AB - BACKGROUND: We designed and evaluated a low-attenuation oral contrast agent for abdominal-pelvic computed tomography (CT). METHODS: In vitro studies, were performed initially to evaluate the imaging characteristics of multiple solutions. These studies resulted in two solutions being compared with the presently accepted oral CT agents of dilute iodinated contrast and water. Ninety eight consecutive subjects already scheduled for routine outpatient abdominal pelvic CT were enrolled. Subjects were randomized to water (n = 30), fiber solution (n = 32), polyethylene glycol (PEG; n = 11), or dilute iodinated solution (DI; n = 25). Examinations were then evaluated for gastric distention, small bowel distention, small bowel wall visualization, and colonic transit. A questionnaire was given to the study subjects for feedback concerning taste and potential side effects from these agents. RESULTS: PEG tended to provide better bowel distention, wall visualization, and colonic transit compared with water, fiber solution, and DI. Areas of statistical significance included: (1) average bowel diameter in the left upper quadrant for water was 17.50 mm, whereas that for PEG was 21.88 mm (p < 0.05); (2) average bowel diameter in the pelvis for water was 14.79 mm, that for fiber was 15.67 mm, and that for PEG was 18.48 mm (p < 0.05); (3) wall visualization was better with PEG than with fiber (p < 0.05); (4) successful transit of contrast to the colon occurred in every subject who received PEG compared with only 20% of those received water and 39% of those who received fiber (p < 0.05). Similar trends for the superiority of PEG over DI were noted, although many of these did not reach statistical significance. CONCLUSION: PEG solution has imaging characteristics related to bowel wall visualization, luminal distention, and colonic transit that make it an effective oral agent for abdominal pelvic CT examination. PMID- 16252140 TI - Intraluminal duodenal diverticulum: MR findings. AB - We report a case of intraluminal duodenal diverticulum with special reference to its magnetic resonance imaging findings. An intraluminal duodenal fluid collection surrounded by a hypointense rim on T2-weighted or magnetic resonance cholangiopancreatographic images, the shape of which is changed by peristalsis, is considered to be almost diagnostic for intraluminal duodenal diverticulum. PMID- 16252141 TI - CT colonography: contrast enhancement of benign and malignant colorectal lesions versus fecal residuals. AB - We retrospectively reviewed the computed tomographic colonographic datasets of 22 patients. Mean attenuation values of benign polyps before and after contrast administration were 30 +/- 15 HU and 90 +/- 18 HU, respectively. Mean attenuation values of colorectal cancer before and after contrast administration were 43 +/- 15 HU and 124 +/- 18 HU, respectively. The mean attenuation value of solid fecal residuals was 43 +/- 15 HU. The difference in attenuation value between precontrast and postcontrast studies of polyps was statistically significant (mean 60 HU, p < 0.01); the same was true for colorectal cancer (mean 81 HU, p < 0.01). The difference between postcontrast density of polyps and cancer with respect to density of solid fecal residuals was statistically significant (p < 0.01). The use of contrast medium could be of help in computed tomographic colonography for discriminating polypoid benign lesions and colorectal cancer from fecal residuals. PMID- 16252142 TI - Bilateral hemorrhagic adrenal cysts in an incomplete form of Beckwith-Wiedemann syndrome: MRI and prenatal US findings. AB - Benign hemorrhagic adrenal cysts are a cause of subdiaphragmatic mass in the fetus and neonate with Beckwith-Wiedemann syndrome. Characteristic imaging features on ultrasonography, color Doppler, and magnetic resonance imaging help differentiate adrenal hemorrhage from neonatal neuroblastoma and help avoid unnecessary surgery in these patients. Bilateral adrenal hemorrhage is self limiting, and spontaneous resolution is the usual outcome. This report presents this rare condition with prenatal ultrasonographic and magnetic resonance imaging findings and reviews the differential diagnosis of neonatal adrenal masses. PMID- 16252143 TI - Vascular involvement secondary to tuberculosis of the abdomen. AB - Although involvement of the abdomen is a common site of extrapulmonary tuberculosis, involvement of abdominal vessels with complications is rare after infections with Mycobacterium tuberculosis (M. tuberculosis). Vascular complications may result from direct involvement by M. tuberculosis of the vascular wall or may occur as a consequence of local spreading from a tuberculous mass. We describe the imaging findings in two patients with proven tuberculosis of the abdomen and significant vascular complications. In one patient, "dry plastic" peritonitis and tuberculous lymphadenopathy at the level of the porta hepatis with subsequent encasement and compression of the portal vein resulted in prestenotic dilatation and varicose formation in the upper abdomen. In the other patient, bulky necrotic tuberculous lymphadenopathy, located at the splenic hilum, resulted in encasement with stenosis of the splenic vessels and subsequent splenic infarction. PMID- 16252145 TI - Virtual CT colectomy by three-dimensional imaging using multidetector-row CT for laparoscopic colorectal surgery. AB - Laparoscopic colorectal surgery has been attracting attention for its capacity to improve the quality of life (QOL) of patients. However, there are disadvantages to this approach, namely, it is difficult to obtain an image of the entire view of the operative field, and organs and lesions cannot be manipulated directly by the surgeon during surgery. For this reason, it takes a relatively large amount of time to ligate vessel, which can vary between patients. Furthermore, vessels and organs can be damaged during lymph nodes dissection under laparoscopic guidance, leading to heavy bleeding that prevents the surgeon from having access to a good view of the operative field. Then, to assess preoperatively the vascular anatomy, we carried out multiphase, contrast-enhanced examinations using multidetector-row CT (MDCT) on patients with colorectal cancer, and prepared the fused image of 3D images of arteries, veins, the colorectum, organs, and tumor. We called the utilization of 3D imaging virtual CT colectomy, which contributed to rapid and safe manipulation of the origins of the arteries and the veins, as well as lymph nodes dissection, without incurring injury to the involved arteries and veins. PMID- 16252144 TI - Radiologic appearance of recurrent ileal Crohn disease. AB - BACKGROUND: Postoperative recurrence of Crohn disease is a common problem. It has been assumed that the radiologic patterns are similar in de novo and recurrent ileal disease, but there has been little in the literature to actually confirm this belief. METHODS: We retrospectively reviewed the small bowel examinations of 105 consecutive patients with a proven diagnosis of Crohn disease: a control group of 47 patients with no prior surgery and a postoperative group of 58 patients with resections. Of the latter, 22 had ileocecal or ileocolonic resections and 36 patients had ileocecal or ileocolonic and extensive enteric resections. We examined the disease sites and compared the disease patterns in both groups. RESULTS: Fifty-six of 58 (97%) postoperative patients had anastomotic recurrences with proximal extension from 3 to 25 cm, with a mean of 10.5 cm; none showed distal disease extension. Two (3%) had enteric recurrences with neoterminal ileal sparing. There were no statistically significant differences in the length of distal/terminal ileal disease and the frequency of skip lesions in de novo and recurrent disease. There were lower frequencies of mucosal thickening, ulceration/ulceronodular mucosa, sacculation, loop separation, sinuses, and masses and a higher frequency of strictures in recurrent disease than in de novo disease. There was also a lower frequency of ulceration or ulceronodular mucosa after extensive resection than after limited resection. CONCLUSION: Postoperative patients with ileal Crohn disease show a marked preponderance for anastomotic recurrence with proximal disease extension. There are significant differences in disease patterns in patients with de novo and recurrent disease. PMID- 16252146 TI - Biliary obstruction secondary to combat-related foreign bodies: report of two cases. AB - We present two cases with combat-related hepatic injury. These patients presented later with obstructive jaundice as seen on the percutaneous transhepatic cholangiography. Combat-related injuries are a rare cause of obstructive jaundice. PMID- 16252147 TI - Kidney length and scarring in children with urinary tract infection: importance of ultrasound scans. AB - BACKGROUND: Many studies have demonstrated that dimercaptosuccinic acid (DMSA) scintigraphy is the most sensitive diagnostic method in the identification of irreversible renal lesions (scars) in children with previous episodes of acute pyelonephritis (APN). This study assessed the reliability of ultrasound in identifying reflux nephropathy in children with acute pyelonephritis with or without vesicoureteric reflux (VUR). METHODS: Eighty children (45 female and 35 male, age range 5 months to 10 years, average age 2 years 1 month) with a positive history for at least one episode of APN participated in this study. All children underwent voiding cystourethrography, DMSA scintigraphy 4 to 8 months after the most recent episode of APN, and an ultrasound test evaluation less than 2 months after DMSA scintigraphy. RESULTS: Voiding cystourethrograms showed VUR in 52 children (68%); 13 of these were bilateral, for a total of 65 refluxing kidney units of the 154 (42%) evaluated; DMSA scintigram was normal for 108 of 154 kidneys (70%). Of the 65 kidneys with VUR, DMSA scintigram displayed normal findings in 29 cases (45%) and pathologic findings in 36 (55%). In the 79 nonrefluxing kidneys, DMSA scintigram was normal in 69 cases (87%). The relative risk of scarring in VUR kidneys is 2.6. The ultrasound study recorded a maximum longitudinal diameter between the 5th and 95th percentiles in 80 of 89 (81%) kidneys without VUR and in 21 of 65 (32%) with VUR. A significant correlation was found between maximum longitudinal diameters and DMSA scintigraphic findings in kidneys with VUR and those without VUR, respectively. CONCLUSION: This study establishes that ultrasound scans, by means of a simple and reproducible measurement technique, maximum longitudinal diameter, have a predictive value with regard to the presence of scars, with few exceptions. This finding, in our opinion, could lead to a decrease in the number of invasive procedures, in particular DMSA scan, in patients with APN. PMID- 16252148 TI - Pancreaticothoracic fistulas: imaging findings in five patients. AB - BACKGROUND: Pancreaticothoracic fistulas are a rare complication of chronic pancreatitis. This study evaluated the imaging findings in patients with pancreaticothoracic fistulas. METHODS: We retrospectively reviewed the medical records and radiographic images in five patients with pancreaticothoracic fistulas diagnosed at our institution during the previous 6 years. Data from radiography, endoscopic retrograde cholangiopancreatography (ERCP), computed tomography (CT), magnetic resonance cholangiopancreatography, and drainage catheter studies were evaluated. RESULTS: Persistent pleural effusions were present on the radiographs of all patients. ERCP was performed in three patients. Contrast material was injected into drainage catheters placed in a pancreatic pseudocyst in one patient and a pancreatic abscess in another patient. Pancreaticothoracic fistulas were demonstrated on all of these studies. One patient had a CT study immediately after ERCP and another patient had a CT study immediately after contrast material was injected through a drainage catheter placed percutaneously in a pseudocyst. CT demonstrated the full extent of the pancreaticothoracic fistula in both patients. CONCLUSION: Pancreaticothoracic fistulas are rare and may require multiple imaging modalities to establish a diagnosis. PMID- 16252149 TI - Treatment of gastrointestinal hemorrhage. AB - BACKGROUND: We assessed the value of selective arteriography in the diagnosis and management of acute gastrointestinal hemorrhage. METHODS: We reviewed the records of 107 consecutive patients who had gastrointestinal hemorrhage and underwent selective arteriography between January 1992 and October 2003: 10 had upper gastrointestinal bleeding, 79 had lower gastrointestinal bleeding, and 18 had varicose bleeding with portal hypertension. Selective embolization was attempted in 15 patients to obtain hemostasis. Angiographic findings were reviewed and prospective reports were compared with the final diagnosis and outcome. RESULTS: Of 129 angiographic studies, 36 correctly revealed the bleeding site and 93 were negative. Extravasation was seen in 24 cases at the level of stomach (n = 2), duodenum (n = 1), small bowel (n = 5), or colon (n = 16). Indirect signs of bleeding sources were identified in 12 patients (stomach in one, small bowel in four, large bowel in four, liver in three). Transcatheter embolization induced definitive hemostasis in 11 of 15 patients (73%), namely in the stomach (n = 2), small bowel (n = 3), colon (n = 7), and liver (n = 3). Three patients required surgery after embolization. CONCLUSION: Abdominal arteriography may localize gastrointestinal bleeding sources in approximately one-third of cases. Selective embolization may provide definitive hemostasis in most instances. PMID- 16252151 TI - Retroanastomotic hernia after gastrojejunostomy: US and CT findings with an emphasis on the whirl sign. AB - BACKGROUND: We reviewed ultrasound (US) and computed tomographic (CT) findings of retroanastomotic hernia and established whether such findings are characteristic of retroanastomotic hernia in patients who have undergone gastrojejunostomy. METHODS: During a recent 10-year period, we encountered 13 consecutive cases of retroanastomotic hernia. Of the patients involved, 11 underwent US, 10 underwent CT, and three underwent small bowel followthrough imaging. The US and CT scans were retrospectively reviewed to determine abnormal findings. Surgical proof was available in all cases. RESULTS: The efferent loop was herniated through the defect created behind the anastomosis in 10 cases, the efferent and afferent loops in two cases, and the afferent loop in one case. Retroanastomotic hernia was prospectively suggested in 12 of these 13 cases. Among the 10 cases of efferent loop herniation, US and CT signs of retroanastomotic hernia included whirling of mesenteric vessels, jejunal loops, and mesentery in the periumbilical abdomen (10 of 10); mural thickening of herniated bowel loops (six of 10); dilatation of herniated bowel loops (four of 10); decreased peristalsis of herniated bowel loops on US (three of nine); and decreased contrast enhancement of herniated bowel loops on CT (one of seven). In one case, US and CT signs of retroanastomotic hernia of the afferent loop included its dilatation and whirling of a short length of afferent loop behind the anastomosis. In two cases, US and CT signs of retroanastomotic hernia of the afferent and efferent loops included findings of afferent and efferent loop herniations. Three of 13 cases had reversible bowel ischemia, and one had bowel necrosis. CONCLUSION: Retroanastomotic hernia is an important condition, and the US and CT findings we have described might suggest its diagnosis. PMID- 16252150 TI - Spontaneous regression of hepatocellular carcinoma with complete necrosis: case report. AB - Spontaneous regression of hepatocellular carcinoma is rare phenomenon. A 74-year old man was found to have a hepatocellular carcinoma with intrahepatic metastases in the lateral segment of the liver. Before surgery, he developed severe cholangitis due to choledocholithiasis and was treated endoscopically. The tumor marker level decreased markedly, and hepatectomy was performed. The resected tumor demonstrated complete necrosis. PMID- 16252152 TI - Cystadenoma of the liver with ovarian-like stroma: case report. AB - Cystadenomas are rare benign tumors with malignant potential. A 36-year-old woman presented with a 20-cm cystic mass in the left lobe of the liver. Surgery confirmed the diagnostic imaging findings. We present a case of cystadenoma of the liver with ovarian-like stroma and its associated radiologic characteristics. PMID- 16252153 TI - Radiofrequency thermal ablation of hepatic tumors: pitfalls and challenges. AB - Image-guided radiofrequency (RF) thermal ablation has been accepted as a promising interventional technique to control unresectable hepatic tumors. One important key to maximize the efficacy of RF ablation is to adhere to the therapeutic guidelines and to avoid preventable pitfalls. There are also several challenges obstructing successful ablation: poor approach path; small sonic window during multiple overlapping ablations; incomplete ablation due to the heat sink effect; subcapsular mass (too exophytic or the mass is adjacent to the gastrointestinal tract); focal residual tumors; too isoechoic or small masses; and mistargeting to adjacent another lesion. Knowledge of these challenges and pitfalls in RF ablation is helpful for a successful ablation. PMID- 16252156 TI - Importance of blood cellular genomic profile in coronary heart disease. AB - Since receptor/transcription factor family especially peroxisome proliferator activated receptors PPARs (alpha, gamma) and liver X receptor alpha (LXRalpha) have been recognized to play crucial role in both lipid metabolism and inflammation, the present study was addressed to explore the interrelationship between blood cellular genomic expression profile, serum lipid levels and severity of coronary heart disease (CHD) in human subjects. Based upon the demographic and laboratory data, the human subjects were divided into 4 groups. Genomic expression profile in the subjects belonging to these groups was determined by measuring the transcriptional expression of genes coding for PPARs (alpha, gamma), CD36, LXRalpha and low density lipoprotein receptor (LDLR) in their blood mononuclear cells. This genomic expression profile was correlated with serum lipid profile as well as with the severity of CHD (revealed by coronary angiography coupled with modified Gensini score) using standard statistical analytical methods. Further in vitro and in vivo effect of statins on such genomic profile was also explored. Although genes coding for PPARs (alpha, gamma), CD36, LDLR showed correlation with the severity of coronary atherosclerosis , blood cellular LXRalpha genomic profile showed conspicuous negative correlation with the severity of coronary atherosclerosis in subjects with or without hypercholesterolemia. This view was further confirmed in experiments directed to understand the effect of statins on the cellular genomic profile of PPARs (alpha, gamma) and LXRalpha. Based on these reported findings, we propose that blood cellular LXRalpha genomic profile has a protective effect against the development of CHD and hence may be of importance in devising synthetic therapeutic drugs for CHD in future. PMID- 16252159 TI - Editorial. PMID- 16252160 TI - Carbohydrate regulation of leaf development: Prolongation of leaf senescence in Rubisco antisense mutants of tobacco. AB - Previous investigations have shown that increased source strength as a result of elevated CO(2) can alter the timing of the phases of change that occur in photosynthetic rates during dicot leaf ontogeny [Miller et al. (1997) Plant Physiol 115: 1195-1200]. To evaluate the converse situation of decreased source strength, we examined leaf development in rbcS antisense mutants of tobacco. These mutants have depressed Rubisco levels and decreased rates of carbohydrate production. We found that antisense leaves are longer-lived than wild type leaves and that this appeared to be due to a prolongation of the senescence phase of development, as monitored by photosynthetic rates, chlorophyll content, and the abundance and activity of Rubisco. Declines in these parameters during leaf ontogeny in both the wild type and mutant plants were generally accompanied by coordinate reductions in the levels of rbcS mRNA and rbcL mRNA, as well as by reductions in chloroplast rRNA, chloroplast DNA and total protein. We suggest that the prolongation of senescence in the antisense leaves is due to an impact of source strength on leaf developmental programming that occurs, at least in part, at the level of transcript abundance of nuclear and chloroplast genes for chloroplast rRNAs and proteins. We hypothesize that plants are capable of sensing a range of source strength conditions to initiate and modulate leaf developmental programming. PMID- 16252161 TI - Photosystem II efficiency and mechanisms of energy dissipation in iron-deficient, field-grown pear trees (Pyrus communis L.). AB - The dark-adapted Photosystem II efficiency of field-grown pear leaves, estimated by the variable to maximum chlorophyll fluorescence ratio, was little affected by moderate and severe iron deficiency. Only extremely iron-deficient leaves showed a decreased Photosystem II efficiency after dark adaptation. Midday depressions in Photosystem II efficiency were still found after short-term dark-adaptation in iron-deficient leaves, indicating that Photosystem II down-regulation occurred when the leaves were illuminated by excessive irradiance. The actual Photosystem II efficiency at steady-state photosynthesis was decreased by iron deficiency both early in the morning and at midday, due to closure of Photosystem II reaction centers and decreases of the intrinsic Photosystem II efficiency. Iron deficiency decreased the amount of light in excess of that which can be used in photosynthesis not only by decreasing absorptance, but also by increasing the relative amount of light dissipated thermally by the Photosystem II antenna. When compared to the controls, iron-deficient pear leaves dissipated thermally up to 20% more of the light absorbed by the Photosystem II, both early in the morning and at midday. At low light iron-deficient leaves with high violaxanthin cycle pigments to chlorophyll ratios had increases in pigment de-epoxidation, non photochemical quenching and thermal dissipation. Our data suggest that DeltapH could be the major factor controlling thermal energy dissipation, and that large (more than 10-fold) changes in the zeaxanthin plus antheraxanthin to chlorophyll molar ratio caused by iron deficiency were associated only to moderate increases in the extent of photoprotection. PMID- 16252163 TI - Effects of azide on the S(2) state EPR signals from Photosystem II. AB - The anion azide, N(3) (-), has been previously found to be an inhibitor of oxygen evolution by Photosystem II (PS II) of higher plants. With respect to chloride activation, azide acts primarily as a competitive inhibitor but uncompetitive inhibition also occurs [Haddy A, Hatchell JA, Kimel RA and Thomas R (1999) Biochemistry 38: 6104-6110]. In this study, the effects of azide on PS II enriched thylakoid membranes were characterized by electron paramagnetic resonance (EPR) spectroscopy. Azide showed two distinguishable effects on the S(2) state EPR signals. In the presence of chloride, which prevented competitive binding, azide suppressed the formation of the multiline and g = 4.1 signals concurrently, indicating that the normal S(2) state was not reached. Signal suppression showed an azide concentration dependence that correlated with the fraction of PS II centers calculated to bind azide at the uncompetitive site, based on the previously determined inhibition constant. No evidence was found for an effect of azide on the Fe(II)Q(A) (-) signals at the concentrations used. This result is consistent with placement of the uncompetitive site on the donor side of PS II as suggested in the previous study. In chloride-depleted PS II-enriched membranes azide and fluoride showed similar effects on the S(2) state EPR signals, including a notable increase and narrowing of the g = 4.1 signal. Comparable effects of other anions have been described previously and apparently take place through the chloride-competitive site. The two azide binding sites described here correlate with the results of other studies of Lewis base inhibitors. PMID- 16252162 TI - Reduction of Q(A) in the dark: Another cause of fluorescence F(o) increases by high temperatures in higher plants. AB - Increases in the chlorophyll fluorescence F(o) (dark level fluorescence) during heat treatments were studied in various higher plants. Besides the dissociation of light-harvesting chlorophyll a/b protein complexes from the reaction center complex of PS II and inactivation of PS II, dark reduction of Q(A) via plastoquinone (PQ) seemed to be related to the F(o) increase at high temperatures. In potato leaves or green tobacco cultured cells, a part of the F(o) increase was quenched by light, reflecting light-induced oxidation of Q(A) ( ) which had been reduced in the dark at high temperatures. Appearance of the F(o) increase due to Q(A) reduction depended on the plant species, and the mechanisms for this are proposed. The reductants seemed to be already present and formed by very brief illumination of the leaves at high temperatures. A ndhB-less mutant of tobacco showed that complex I type NAD(P)H dehydrogenase is not involved in the heat-induced reduction of Q(A). Quite strong inhibition of the Q(A) reduction by diphenyleneiodonium suggests that a flavoenzyme is one of the electron mediator to PQ from the reductant in the stroma. Reversibility of the heat-induced Q(A) reduction suggests that an enzyme(s) involved is activated at high temperatures and mostly returns to an inactive form at room temperature (25 degrees C). PMID- 16252164 TI - An FTIR study on the structure of the oxygen-evolving Mn-cluster of Photosystem II in different spin forms of the S(2) state. AB - The S(2) state of the oxygen-evolving Mn-cluster of Photosystem II (PS II) is known to have different forms that exhibit the g =2 multiline and g = 4.1 EPR signals. These two spin forms are interconvertible at > 200 K and the relative amplitudes of the two signals are dependent on the species of cryoprotectant and alcohol contained in the medium. Also, it was recently found that the mutiline form can be converted to the g = 4.1 form by absorption of near-infrared light by the Mn-cluster itself at around 150 K [Boussac et al. (1996) Biochemistry 35: 6984-6989]. We have used light-induced Fourier transform infrared (FTIR) difference spectroscopy to study the structural difference in these two S(2) forms. FTIR difference spectra for S(2)/S(1) as well as for S(2)Q(A) (-)/S(1)Q(A) measured at cryogenic temperatures using PS II membranes in the presence of various cryoprotectants, and monohydric alcohols did not show any specific differences except for intensities of amide I bands, which were larger when ethylene glycol or glycerol was present in addition to sucrose. This result was interpreted due to more flexible movement of the protein backbones upon S(2) formation with a higher cryoprotectant content. Light-induced difference spectra measured at 150 K using either blue light without near-infrared light or red plus near-infrared light also did not show any detectable difference. In addition, a different spectrum upon near-infrared illumination at 150 K of the PS II sample in which the S(2) state had been photogenerated at 200 K exhibited no meaningful signals. These results indicate that the two S(2) forms that give rise to the multiline and g = 4.1 signals have only minor differences, if any, in the structures of amino-acid ligands and polypeptide backbones. This conclusion suggests that conversion between the two spin states is caused by a spin-state transition in the Mn(III) ion rather than valence swapping within the Mn-cluster that would considerably affect the vibrations of ligands. PMID- 16252165 TI - Acclimation of photosynthesis to temperature in eight cool and warm climate herbaceous C(3) species: Temperature dependence of parameters of a biochemical photosynthesis model. AB - To determine how parameters of a Farquhar-type photosynthesis model varied with measurement temperature and with growth temperature, eight cool and warm climate herbaceous crop and weed species were grown at 15 and 25 degrees C and single leaf carbon dioxide and water vapor exchange rates were measured over the range of 15 - 35 degrees C. Photosynthetic parameters examined were the initial slope of the response of assimilation rate (A) to substomatal carbon dioxide concentration (C(i)), A at high C(i), and stomatal conductance. The first two measurements allow calculation of V(Cmax), the maximum rate of carboxylation of ribulose bisphosphate carboxylase and J(max), the maximum rate of photosynthetic electron transport, of Farquhar-type photosynthesis models. In all species, stomatal conductance increased exponentially with temperature over the whole range of 15 - 35 degrees C, even when A decreased at high measurement temperature. There were larger increases in conductance over this temperature range in the warm climate species (4.3 x) than in the cool climate species (2.5 x). The initial slope of A vs. C(i) exhibited an optimum temperature which ranged from 20 to 30 degrees C. There was a larger increase in the optimum temperature of the initial slope at the warmer growth temperature in the cool climate species than in the warm climate species. The optimum temperature for A at high C(i) ranged from 25 to 30 degrees C among species, but changed little with growth temperature. The absolute values of both the initial slope of A vs. C(i) and A at high C(i) were increased about 10% by growth at the warmer temperature in the warm climate species, and decreased about 20% in the cool climate species. The ratio of J(max) - V(Cmax) normalized to 20 degrees C varied by more than a factor of 2 across species and growth temperatures, but differences in the temperature response of photosynthesis were more related to variation in the temperature dependencies of J(max) and V(Cmax) than to the ratio of their normalized values. PMID- 16252166 TI - Occurrence of C(3) and C(4) photosynthesis in cotyledons and leaves of Salsola species (Chenopodiaceae). AB - Most species of the genus Salsola (Chenopodiaceae) that have been examined exhibit C(4) photosynthesis in leaves. Four Salsola species from Central Asia were investigated in this study to determine the structural and functional relationships in photosynthesis of cotyledons compared to leaves, using anatomical (Kranz versus non-Kranz anatomy, chloroplast ultrastructure) and biochemical (activities of photosynthetic enzymes of the C(3) and C(4) pathways, (14)C labeling of primary photosynthesis products and (13)C/(12)C carbon isotope fractionation) criteria. The species included S. paulsenii from section Salsola, S. richteri from section Coccosalsola, S. laricina from section Caroxylon, and S. gemmascens from section Malpigipila. The results show that all four species have a C(4) type of photosynthesis in leaves with a Salsoloid type Kranz anatomy, whereas both C(3) and C(4) types of photosynthesis were found in cotyledons. S. paulsenii and S. richteri have NADP- (NADP-ME) C(4) type biochemistry with Salsoloid Kranz anatomy in both leaves and cotyledons. In S. laricina, both cotyledons and leaves have NAD-malic enzyme (NAD-ME) C(4) type photosynthesis; however, while the leaves have Salsoloid type Kranz anatomy, cotyledons have Atriplicoid type Kranz anatomy. In S. gemmascens, cotyledons exhibit C(3) type photosynthesis, while leaves perform NAD-ME type photosynthesis. Since the four species studied belong to different Salsola sections, this suggests that differences in photosynthetic types of leaves and cotyledons may be used as a basis or studies of the origin and evolution of C(4) photosynthesis in the family Chenopodiaceae. PMID- 16252167 TI - Characterization of the Porphyridium cruentum Chl a-binding LHC by in vitro reconstitution: LHCaR1 binds 8 Chl a molecules and proportionately more carotenoids than CAB proteins. AB - The Porphyridium cruentum light harvesting complex (LHC) binds Chl a, zeaxanthin and beta-carotene and comprises at least 6 polypeptides of a multigene family. We describe the first in vitro reconstitution of a red algal light-harvesting protein (LHCaR1) with Chl a/carotenoid extracts from P. cruentum. The reconstituted pigment complex (rLHCaR1) is spectrally similar to the native LHC I, with an absorption maximum at 670 nm, a 77 K fluorescence emission peak at 677 nm (ex. 440 nm), and similar circular dichroism spectra. Molar ratios of 4.0 zeaxanthin, 0.3 beta-carotene and 8.2 Chl a per polypeptide for rLHCaR1 are similar to those of the native LHC I complex (3.1 zeaxanthin, 0.5 beta-carotene, 8.5 Chl a). The binding of 8 Chl a molecules per apoprotein is consistent with 8 putative Chl-binding sites in the predicted transmembrane helices of LHCaR1. Two of the putative Chl a binding sites (helix 2) in LHCaR1 were assigned to Chl b in Chl a/b-binding (CAB) LHC II [Kuhlbrandt et al. (1994) Nature 367: 614-21]. This suggests either that discrimination for binding of Chl a or Chl b is not very specific at these sites or that specificity of binding sites evolved separately in CAB proteins. LHCaR1 can be reconstituted with varying ratios of carotenoids, consistent with our previous observation that the carotenoid to Chl ratio is substantially higher in P. cruentum grown under high irradiance. Also notable is that zeaxanthin does not act as an accessory light-harvesting pigment, even though it is highly likely that it occupies the position assigned to lutein in the CAB LHCs. PMID- 16252168 TI - Photosynthetic acclimation of higher plants to growth in fluctuating light environments. AB - This paper describes a study into the potential of plants to acclimate to light environments that fluctuate over time periods between 15 min and 3 h. Plants of Arabidopsis thaliana (L.) Heynh., Digitalis purpurea L. and Silene dioica (L.) Clairv. were grown at an irradiance 100 mumol m(-2) s(-1). After 4-6 weeks, they were transferred to light regimes that fluctuated between 100 and either 475 or 810 mumol m(-2) s(-1), in a regular cycle, for 7 days. Plants were shown, in most cases, to be able to undergo photosynthetic acclimation under such conditions, increasing maximum photosynthetic rate. The extent of acclimation varied between species. A more detailed study with S. dioica showed that this acclimation involved changes in both Rubisco protein and cytochrome f content, with only marginal changes in pigment content and composition. Acclimation to fluctuating light, at the protein level, did not fully reflect the acclimation to continuous high light - Rubisco protein increased more than would be expected from the mean irradiance, but less than expected from the high irradiance; cytochrome f increased when neither the mean nor the high irradiance would be expected to induce an increase. PMID- 16252169 TI - Obituary: Remembering George Cheniae, who never compromised his high standards of science. PMID- 16252170 TI - Carotenoids and bacterial photosynthesis: The story so far... AB - This concise review describes the current status of research into how carotenoids function in bacterial photosynthesis. This is illustrated by reference to very recent studies on both the photoprotective and antenna functions of carotenoids. The major remaining open questions on the detailed molecular mechanisms involved in these reactions are highlighted. PMID- 16252171 TI - The protochlorophyllide-chlorophyllide cycle. AB - The protochlorophyllide-chlorophyllide cycle was introduced by Sironval [1981, In: Akoyunoglou G (ed) Photosynthesis, Vol 5: Chloroplast Development, pp 3-14] to describe the interconvertion of the Pchlide and Chlide spectral forms occurring when nonilluminated leaves are irradiated for the first time. Since this original publication important progresses have been made at the physiological, biochemical and molecular levels. The understanding of the cycle and of its regulation are critical for the understanding of chlorophyll synthesis in photosynthetic organisms. In this contribution, I present an update version of the cycle in tissues illuminated for the first time, in greening and in green leaves, and in photosynthetic organisms, which are able to synthesize chlorophyll in the dark as well. To the descriptive part, an assay on the regulation of the cycle is added. PMID- 16252172 TI - Heterogeneity of Photosystem I reaction centers in barley leaves as related to the donation from stromal reductants. AB - The light-response curves of P700 oxidation and time-resolved kinetics of P700(+) dark re-reduction were studied in barley leaves using absorbance changes at 820 nm. Leaves were exposed to 45 degrees C and treated with either diuron or diuron plus methyl viologen (MV) to prevent linear electron flow from PS II to PS I and ferredoxin-dependent cyclic electron flow around PS I. Under those conditions, P700(+) could accept electrons solely from soluble stromal reductants. P700 was oxidized under weak far-red light in leaves treated with diuron plus MV, while identical illumination was nearly ineffective in diuron-treated leaves in the absence of MV. When heat-exposed leaves were briefly illuminated with strong far red light, which completely oxidized P700, the kinetics of P700(+) dark reduction was fitted by a single exponential term with half-time of about 40 ms. However, two first-order kinetic components of electron flow to P700(+) (fast and slow) were found after prolonged leaf irradiation. The light-induced modulation of the kinetics of P700(+) dark reduction was reversed following dark adaptation. The fast component (half time of 80-90 ms) was 1.5 larger than the slow one (half time of about 1 s). No kinetic competition occurred between two pathways of electron donation to P700(+) from stromal reductants. This suggests the presence of two different populations of PS I. PMID- 16252173 TI - A comparative study of the thermal stability of plastocyanin, cytochrome c(6) and Photosystem I in thermophilic and mesophilic cyanobacteria. AB - Cytochrome c(6) (Cyt) from the thermophilic cyanobacterium Phormidium laminosum has been purified and characterized. It is a mildly acidic protein, with physicochemical properties very similar to those of plastocyanin (Pc). This is in agreement with the functional interchangeability of the two metalloproteins as electron donors to Photosystem I (PS I). The kinetic analyses of the interaction of Pc and Cyt with Photosystem I show that both metalloproteins reduce PS I with similar efficiencies, according to an oriented collisional kinetic model involving repulsive electrostatic interactions. The thermostability study of the Phormidium Pc/PS I system compared with those from mesophilic cyanobacteria (Synechocystis, Anabaena and Pseudanabaena) reveals that Pc is the partner limiting the thermostability of the Phormidium couple. The cross-reactions between Pc and PS I from different organisms demonstrate not only that Phormidium Pc enhances the stability of the Pc/PS I system using PS I from mesophilic cyanobacteria, but also that Phormidium PS I possesses a higher thermostability than the other photosystems. PMID- 16252174 TI - Inhibition by light of CO(2) evolution from dark respiration: Comparison of two gas exchange methods. AB - Two approaches to determine the fraction (mu) of mitochondrial respiration sustained during illumination by measuring CO(2) gas exchange are compared. In single leaves, the respiration rate in the light ('day respiration' rate R(d)) is determined as the ordinate of the intersection point of A-c(i) curves at various photon flux densities and compared with the CO(2) evolution rate in darkness ('night respiration' rate R(n)). Alternatively, using leaves with varying values of CO(2) compensation concentration (Gamma), intracellular resistance (r(i)) and R(n), an average number for mu can be derived from the linear regression between Gamma and the product r(i)cR(n). Both methods also result in a number c* for that intercellular CO(2) concentration at which net CO(2) uptake rate is equal to R(d). c* is an approximate value of the photocompensation point Gamma* (Gamma in the absence of mitochondrial respiration), which is related to the CO(2)/O(2) specificity factor of Rubisco S(c/o). The presuppositions and limitations for application of both approaches are discussed. In leaves of Nicotiana tabacum, at 22 degrees C, single leaf measurements resulted in mean values of mu = 0.71 and c(*) = 34 mumol mol(-1). At the photosynthetically active photon flux density of 960 mumol quanta m(-2) s(-1), nearly the same numbers were derived from the linear relationship between Gamma and r(i)cR(n). c* and R(d) determined by single leaf measurements varied between 31 and 41 mumol mol(-1) and between 0.37 and 1.22 mumol m(-2) s(-1), respectively. A highly significant negative correlation between c* and R(d) was found. From the regression equation we obtained estimates for Gamma* (39 mumol mol(-1)), S(c/o) (96.5 mol mol(-1)) and the mesophyll CO(2) transfer resistance (7.0 mol(-1) m(2) s). PMID- 16252175 TI - Light acclimation maintains the redox state of the PS II electron acceptor Q(A) within a narrow range over a broad range of light intensities. AB - Chrysanthemum inducum-hybrid 'Coral Charm', Hibiscus rosa-sinensis L. 'Cairo Red' and Spathiphyllum wallisii Regel 'Petit' were grown in natural light in a greenhouse at three levels of irradiance using permanent shade screens. Light acclimation of photosynthesis was characterized using modulated chlorophyll a fluorescence of intact leaves. A close correlation was found between the degree of reduction of the primary electron acceptor Q(A) of Photosystem II (PS II) approximated as the fluorescence parameter 1-q(P), and light acclimation. The action range of 1-q(P) was 0-0.4 from darkness to full irradiance around noon, within the respective light treatments in the greenhouse, indicating that most PS II reaction centres were kept open. In general, the index for electron transport (ETR) measured by chlorophyll fluorescence was higher for high-light (HL) than intermediate-(IL) and low-light (LL) grown plants. However, HL Chrysanthemum showed 40% higher ETR than HL Hibiscus at light saturation, despite identical redox states of Q(A). The light acclimation of the non-radiative dissipation of excess energy in the antenna, NPQ, varied considerably between the species. However, when normalized against q(P), a strong negative correlation was found between thermal dissipation and ETR measured by chlorophyll fluorescence. To be able to accommodate a high flux of electrons through PS II, the plants with the highest light-saturated ETR had the lowest NPQ/q(P). The possibility of using chlorophyll fluorescence for quantification of the energy balance between energy input and utilization in PS II in intact leaves is discussed. PMID- 16252176 TI - Ligation-mediated suppression-PCR as a powerful tool to analyse nuclear gene sequences in the green alga Chlamydomonas reinhardtii. AB - To improve the analysis of unknown flanking DNA sequences adjacent to known sequences in nuclear genomes of photoautotrophic eukaryotic organisms, we established the technique of ligation-mediated suppression-PCR (LMS-PCR) in the green alga Chlamydomonas reinhardtii for (1) walking from a specific nuclear insertion fragment of random knockout mutants into the unknown flanking DNA sequence to identify and analyse disrupted genomic DNA regions and for (2) walking from highly conserved DNA regions derived from known gene iso-forms into flanking DNA sequences to identify new members of protein families. The feasibility of LMS-PCR for these applications was successfully demonstrated in two different approaches. The first resulted in the identification of a genomic DNA fragment flanking a nuclear insertion vector in a random knockout mutant whose phenotype was characterised by its inability to perform functional LHC state transitions. The second approach targeted the cab gene family. An oligonucleotide of a cabII gene, derived from a highly conserved region, was used to identify potential cab gene regions in the nuclear genome of Chlamydomonas. LMS-PCR combined with 3' rapid amplification of cDNA ends (3' RACE) and a PCR based screening of a cDNA library resulted in the identification of the new cabII gene lhcb4. Both results clearly indicate that LMS-PCR is a powerful tool for the identification of flanking DNA sequences in the nuclear genome of Chlamydomonas reinhardtii. PMID- 16252178 TI - Announcement: Calvin and Hill prizes: 2001. PMID- 16252177 TI - Re-identification of the N-terminal amino acid residue and its modification of +/ bbeta-polypeptide of light-harvesting complex I from Rhodospirillum rubrum. PMID- 16252181 TI - Evaluation of oxidant stress in Wilson's disease and non-Wilsonian chronic liver disease in childhood. AB - BACKGROUND/AIMS: Oxygen free radicals have an important role in the pathogenesis of acute and chronic liver disease. Free radical formation and oxidative damage, probably mediated with copper accumulation, are important in Wilson's disease pathogenesis. This study was performed to determine if accumulating copper in Wilson's disease is a cause of further oxidant stress compared to non-Wilsonian liver disease. METHODS: In this study, we investigated plasma malondialdehyde and nitric oxide levels to estimate the oxidant stress and total antioxidant capacity and vitamin E/cholesterol, vitamin C and beta-carotene levels to estimate the antioxidant status of patients. The groups investigated included 24 patients with Wilson's disease (group I), 25 patients with non-Wilsonian chronic liver disease (group II) and 23 healthy controls (group III). Wilson's disease and non-Wilson's disease patients were divided into subgroups according to disease stage (i.e. chronic hepatitis and cirrhosis) and all parameters were compared between subgroups and controls. RESULTS: Malondialdehyde and nitric oxide levels were higher than controls in groups I and II (p=0.013, p=0.01), but these levels did not differ between the Wilson's disease and non-Wilson's disease groups. The parameters were also evaluated with respect to the disease stage (i.e. chronic hepatitis and cirrhosis), and there was no difference between groups I and II. Although malondialdehyde and nitric oxide levels were significantly different between both disease stage groups and the controls, we observed decreased vitamin C and beta-carotene levels only in cirrhosis stage (p=0.01, p=0.01). CONCLUSIONS: We observed the presence of oxidant stress unrelated to the etiology of the liver disorder in our study. Deficiency of the major antioxidants, vitamin C and beta carotene, develops as the disease stage advances from chronic hepatitis to cirrhosis. PMID- 16252180 TI - Apoptosis and fibrosis in non-alcoholic fatty liver disease. AB - Nonalcoholic fatty liver disease is becoming an increasingly common medical problem in the developed countries which, unfortunately, still is associated with the lack of any effective treatment. However, recent data favor a model in which a pathologically increased rate of hepatocytic apoptosis and the subsequent induction and upregulation of inflammation and fibrosis in the liver provide both a rationale for the pathogenesis of nonalcoholic fatty liver disease, as well as a clue for designing first effective therapeutic strategies. In order to illuminate this context, this article focuses on the pathogenesis and possible new therapeutic options in nonalcoholic fatty liver disease. PMID- 16252182 TI - P21 expression and its relation to disease activity and hepatocyte proliferation in chronic hepatitis B virus infection. AB - BACKGROUND/AIMS: P21 protein, a cell cycle regulatory protein expressed in the liver, acts as an inhibitor of cyclin dependent kinase and prevents progression of the cell cycle. In the present study, our aim was to investigate the relationships between P21 protein expression and hepatocyte proliferation, hepatitis B virus replication, and hepatitis activity. METHODS: A total of 66 patients with chronic hepatitis B without cirrhosis were included in the study. These patients were evaluated in three different groups according to the degree of viral replication and the disease activity. Group 1: HBeAg-positive patients with active liver disease and with viral replication, group 2: HBeAg-negative patients with active liver disease and with viral replication, and group 3: HBeAg negative inactive carriers. P21 and proliferating cell nuclear antigen were immunohistochemically stained and a labeling index was calculated for each protein. RESULTS: A total of 32 (48.4%) patients were positive for nuclear P21 expression. All three groups had a similar P21 index. proliferating cell nuclear antigen-labeling index, hepatit B virus DNA levels, ALT levels, and HAI scores were not different in patients with and without P21 staining. Spearman's correlation analysis found no correlation between P21 staining and ALT and hepatit B virus DNA levels, HAI score and proliferating cell nuclear antigen labeling index. CONCLUSIONS: These results suggest that the pattern of P21 expression is not associated with histological activity, hepatocyte proliferation and virus replication in patients with well-compensated chronic hepatitis B. PMID- 16252183 TI - Chronic hepatitis delta virus infection in Van region of eastern Turkey. AB - BACKGROUND/AIMS: Hepatitis delta virus infection is an important cause of liver morbidity and mortality worldwide. In Eastern Turkey, hepatitis B virus infection is the major cause of chronic liver diseases. We aimed to research the role of hepatitis delta virus infection in chronic liver diseases related to hepatitis B virus infection in the Van region of Eastern Turkey. METHODS: Serological markers of hepatitis B virus and hepatitis delta virus infection [HBsAg, HbeAg, Anti-HBe and Anti- hepatitis delta virus total (IgM+IgG)] were determined by ELISA test in patients with chronic hepatitis and cirrhosis. Serum hepatitis B virus DNA was determined by polymerase chain reaction (PCR) method in chronic hepatitis B patients. RESULTS: Hepatitis delta virus infection was detected in 5% (7/138) of asymptomatic hepatitis B virus carriers, in 16% (24/148) of chronic hepatitis B patients and in 45% (34/75) of cirrhotic hepatitis B virus patients. hepatitis delta virus infection showed a three-fold increase in chronic hepatitis (p<0.01) and nine-fold increase in cirrhosis (p<0.001) compared to hepatitis delta virus carriers. Also, it was three times more frequent in cirrhosis (p<0.001) compared to chronic hepatitis. Chronic hepatitis delta virus infection was equally distributed between sexes in patients with chronic hepatitis B virus infection, whereas chronic hepatitis B virus infection alone was three times more frequent in males (p<0.001). Mean ages of hepatitis delta virus carriers, chronic hepatitis D and hepatitis delta virus cirrhosis patients were 30.7+/-8 (14-65), 36+/-13 (19-70) and 44 +/-16 (25-55), respectively. CONCLUSIONS: The higher prevalence of hepatitis delta virus infection in more severe form of hepatitis B virus infection suggests that hepatitis delta virus infection increases the severity of chronic hepatitis B virus infection in the Van region. hepatitis delta virus infection remains a second major cause of chronic liver diseases in Eastern Turkey in spite of its decreasing prevalence in Western countries and in Western Turkey. PMID- 16252184 TI - Treatment of biliary leakages after cholecystectomy and importance of stricture development in the main bile duct injury. AB - BACKGROUND/AIMS: Endoscopic treatment of biliary leakages after cholecystectomy, though widely accepted, has some restrictions. The efficacy and safety of endoscopic treatments in this patient group are evaluated in this study, and the problem of biliary stricture development in time after biliary ductal injuries is also emphasized. METHODS: Seventy-four patients (20 male, 54 female, mean age 50.9+/-21 years) referred for ERCP between 1992-2002 were included in the study. Minor leakages (cystic duct leaks, accessory bile duct leaks) were managed by nasobiliary drainage +/- endoscopic sphincterotomy; major leakages were managed by nasobiliary drainage +/- endoscopic sphincterotomy +/- stenting. RESULTS: Twenty-seven patients with cystic duct leaks and 6 patients with accessory bile duct leaks were successfully treated with nasobiliary drainage. Endoscopic treatment could not be performed on patients with total bile duct obstruction (7 patients) and aberrant bile duct injury (7 patients). All leakages from main bile ducts were closed (27 patients). Six of 27 patients had strictures at the beginning and they were treated by stenting. Twenty-one patients had no strictures at the beginning. Eight of 21 were treated by stenting and only 1 of them developed biliary stricture. Seven of 13 patients who had been treated by nasobiliary drainage developed biliary strictures. There were no mortalities due to procedure. CONCLUSIONS: ERCP is an effective and safe method for diagnosis and management of bile leakages after cholecystectomy. Stricture development in the main bile duct leakages was an important complication. PMID- 16252185 TI - Can lansoprazole, amoxicillin, and clarithromycin combination still be used as a first-line therapy for eradication of helicobacter pylori? AB - BACKGROUND/AIMS: To determine H. pylori eradication rate with lansoprazole amoxicillin-clarithromycin treatment regimen, which is the most frequently used as first-line therapy, in the Kirikkale region. METHODS: One hundred and five patients (44 male, 61 female) with H. pylori infection were included in the study. Patients were divided into two groups based on the endoscopic findings: non-ulcer dyspepsia (n=84, 31 male, 53 female) and acute gastric or duodenal ulcer (n=21, 13 male, 8 female) groups. The diagnosis of H. pylori infection was confirmed if both the urease test and histological examination, which were performed on endoscopic biopsies, were positive. Lansoprazole 30 mg, amoxicillin 1 g, and clarithromycin 500 mg were given twice daily for 14 days to all patients. Endoscopic biopsies were repeated for the evaluation of eradication three months after the treatment. RESULTS: Ninety-six patients completed the study. Eradication rates were found to be 45.8% (44 of 96) in all patients, 42.1% (32 of 76 patients) in the non-ulcer dyspepsia group and 60% (12 of 20 patients) in the gastric or duodenal ulcer group for per protocol analysis, and the difference between non-ulcer dyspepsia and gastric or duodenal ulcer groups was not statistically significant (p=0.208). CONCLUSIONS: Lansoprazole-amoxicillin clarithromycin treatment regimen, the most frequently preferred regimen in H. pylori eradication, is ineffective in our region. The low eradication rates observed with lansoprazole-amoxicillin-clarithromycin, at least in our region, bring into question its use as a first-line therapy. The use of alternative treatment protocols or antibiotic susceptibility test before the treatment may be helpful in achieving successful eradication with first-line therapy. PMID- 16252186 TI - Complete resolution of transplantation-associated thrombotic microangiopathy and hepatic veno-occlusive disease by defibrotide and plasma exchange. AB - Transplantation-associated thrombotic microangiopathy has been associated with significantly reduced survival following allogeneic bone marrow transplantation. We describe here the course of Transplantation-associated thrombotic microangiopathy and hepatic veno-occlusive disease, and response to plasma exchange therapy. A 19-year-old male patient underwent hematopoietic stem cell transplantation (HSCT) from his HLA-matched brother for lymphoblastic lymphoma in the first complete remission. Transplantation-associated thrombotic microangiopathy was diagnosed 17 days after transplantation. At that time, neurological abnormalities were not present. Cyclosporin A (CsA) was discontinued. Hematological stabilization was recorded. On day +20, abdominal distention, painful hepatomegaly and ascites complicated the clinical picture. With a high hepatic venous pressure gradient (18mmH20), veno-occlusive disease of the liver was diagnosed and defibrotide was started, which resulted in a dramatic cessation of pain and increase in urinary output. However, transplantation associated thrombotic microangiopathy-related symptoms progressed and plasma exchange was instituted, which resulted in worsening of veno-occlusive disease symptoms. He was referred to the Intensive Care Unit due to respiratory compromise and was intubated. Plasma exchange was continued in order after hemofiltration. In three days, fever resolved, hemofiltration could be stopped, and ventilator dependence ended. After 19 aphereses, serum LDH level returned to normal and schistocytes were minimal on microscopic examination of the blood film. Platelet count increase was more gradual. Plasma exchange was discontinued. On the 40th day of defibrotide, all symptoms related with veno-occlusive disease were resolved and defibrotide was stopped. We think that our case is important to establish the relation and management strategy of these two small vessel complications of HSCT. PMID- 16252187 TI - Spontaneous bacterial peritonitis due to Brucella Melitensis in a cirrhotic patient. AB - Spontaneous bacterial peritonitis is a well-known entity, with a reported incidence of 15-20% in advanced cirrhotic patients. Escherichia coli and Klebsiella pneumoniae are the most common causes of spontaneous bacterial peritonitis; Brucella is extremely rare. We aimed to present one case of such a rare condition in a cirrhotic patient who also had hepatocellular carcinoma. Routine laboratory tests, abdominal ultrasonography and peritoneal fluid examinations were studied in a cirrhotic patient with ascites. Peritoneal fluid white blood cell count was 1300/mm3, with lymphocyte predominance (80%). Peritoneal fluid and blood culture both yielded Brucella melitensis. The patient also had a mass in the right lobe of the liver confirmed as hepatocellular carcinoma by biopsy. Brucella should be suspected as a cause of spontaneous bacterial peritonitis in cirrhotic patients with no response to standard spontaneous bacterial peritonitis treatments and with immunodeficiency such as hepatocellular carcinoma. PMID- 16252188 TI - Scintigraphic diagnosis of protein-losing enteropathy secondary to amyloidosis. AB - Protein-losing enteropathy is an uncommon syndrome of excessive loss of protein via the gastrointestinal mucosa. 99mTc-dextran scintigraphy was performed on a 42 year-old woman with protein-losing enteropathy. She had secondary amyloidosis due to rheumatoid arthritis. Abnormal leakage of the radiotracer was observed in the mid-abdominal region suggesting the site of protein loss. It is concluded that 99mTc-dextran scintigraphy is useful as a noninvasive and simple test for the imaging and confirmation of diagnosis in protein-losing enteropathy. PMID- 16252189 TI - Restorative proctocolectomy for familial adenomatous polyposis coexisting with colorectal cancer. AB - The aim of this study was to assess whether restorative proctocolectomy was suitable as an initial procedure for selected familial adenomatous polyposis patients with coexisting colorectal cancer. Six malignancy patients who underwent restorative proctocolectomy for familial adenomatous polyposis were reviewed. At the time of restorative proctocolectomy, cancer was not suspected in four patients. The two remaining familial adenomatous polyposis patients had a known associated colorectal cancer. Operative procedures, pathological findings, complications and long-term follow-up were evaluated. All patients were Stage I II cancers. There was no mortality or pouch failure. No evidence of tumor recurrence was found and all the patients are still alive and disease-free in follow-up (mean 28 months). As an initial procedure, restorative proctocolectomy for patients with familial adenomatous polyposis with coexisting Stage I-II colorectal cancer seems to be a dependable procedure. PMID- 16252190 TI - Colonic obstruction due to rectal endometriosis: report of a case. AB - Although endometriosis is a common disease in women of childbearing age, intestinal endometriosis is unusual and may cause clinically significant complications. We report a 46-year-old woman with rectal endometriosis who presented with intestinal obstruction. She was operated on with a preoperative diagnosis of malignancy. The diagnosis of endometriosis was made only after histological examination of the resected specimen. Intestinal endometriosis has a diverse clinical spectrum, with nonspecific features in many patients. In female patients who have unexplained digestive complaints, endometriosis should also be considered in the differential diagnosis. PMID- 16252191 TI - A case of visceral leishmaniasis misdiagnosed as autoimmune hepatitis. PMID- 16252192 TI - Adult intussusception in two cases. PMID- 16252193 TI - Radiological evaluation of internal abdominal hernias. AB - An internal abdominal herniation is the protrusion of a viscus through a normal or abnormal mesenteric or peritoneal aperture. Internal abdominal herniations can either be acquired through a trauma or surgical procedure, or constitutional and related to congenital peritoneal defects. Paraduodenal hernias are the most common type of internal abdominal hernias, accounting for over one-half of reported cases, and thus are a significant clinical entity. Other internal hernias include pericecal, transmesenteric, transomental, intersigmoid, supravesical hernias and herniation through the foramen of Winslow. Because internal abdominal herniations are rare, their diagnosis remains a challenge for both the clinician and the radiologist. Symptoms of internal abdominal herniations are nonspecific. We present our experience with the radiological evaluation of internal abdominal herniations and review the main radiologic findings on barium as well as computed tomography studies. PMID- 16252194 TI - Assessment of lipid peroxidation and antioxidant capacity in non-alcoholic fatty liver disease. AB - BACKGROUND/AIMS: Lipid peroxidation/oxidative stress and/or endotoxin-induced cytokine release are implicated in the non-alcoholic fatty liver disease pathogenesis. Studies in the literature are experimental in nature and are based on histopathological evaluation and peripheral blood findings. In this study, we aimed to investigate the relationships between lipid peroxidation and antioxidant capacity in non-alcoholic fatty liver disease. METHODS: Twenty-six patients with an ethanol consumption of less than 20 g/day who were diagnosed ultrasonographically and histopathologically as non-alcoholic fatty liver disease and 16 healthy control subjects with normal ultrasonographical findings were included in the study. All viral and autoimmune markers in patient and control groups included in the study were negative. Non-alcoholic steatohepatitis samples obtained by fine needle aspiration were evaluated according to Brunt et al. The levels of glutathione, catalase, superoxide dismutase, and malonyldialdehyde in peripheral blood and liver biopsy samples were measured. RESULTS: Of patients with non-alcoholic fatty liver disease, 17 (65%) were detected to have mild, 7 (27%) moderate and 2 (8%) severe steatosis; portal inflammation was found in 17 patients (65%) and stage I fibrosis in 21 patients (80%). Minimal lobular inflammation was observed in all patients. In the patient group, the levels of erythrocytic glutathione, catalase, and superoxide dismutase were significantly lower but malonyldialdehyde levels were higher compared to the control group. It was revealed that hepatocytic reduced glutathione, catalase, and malonyldialdehyde levels were not correlated with peripheral blood levels, but there was a positive correlation between liver malonyldialdehyde level and liver reduced glutathione level. Plasma malonyldialdehyde level and liver glutathione had a negative correlation. CONCLUSIONS: It was discovered that lipid peroxidation and antioxidant capacity suppression due to its overuse were important in the non-alcoholic fatty liver disease pathogenesis, but antioxidant capacity was maintained well at tissue level in the early stages of the disease. Furthermore, it was identified that tissue lipid peroxidation and changes in antioxidant capacity were not reflected in the peripheral blood to the same extent. PMID- 16252195 TI - Wilson disease in southern Iran. AB - BACKGROUND/AIMS: Wilson disease is an autosomal recessive disorder of the brain, liver and cornea. It is fatal if left untreated. This descriptive study attempted to identify the demographic, clinical, and biochemical features of Wilson disease in Fars Province, Southern Iran. METHODS: All the patients with Wilson disease who were admitted to Nemazee Hospital, Shiraz, Iran, from 1990 till 2004 were included in this cross-sectional descriptive study. This is the only hospital in Southern Iran which has pediatric and adult Gastroenterology Wards. Statistical analyses were done by t-test. RESULTS: In total, 111 patients were studied (65 males, 46 females). The mean age was 11+/-7 years; the youngest was three years old. The most common manifestations were hepatic (83.8%), neurological (24.3%) and psychological (23.4%) signs and symptoms. The most common biochemical abnormalities were increased urinary copper (91.4%), increased prothrombin time (84.5%), increased liver enzymes (77-89%), decreased serum ceruloplasmin (75.5%), hyperbilirubinemia (67%) and anemia (62.4%). Family history was positive in 36% of the patients. CONCLUSIONS: In this study, sex ratio (M/F) was greater than one, similar to other studies. Age range was similar to other Asian studies, but less than observed in European and American studies. Clinical manifestations of Wilson disease in our patients were similar to the studies with similar age ranges. Biochemical abnormalities were sometimes different in our patients in comparison to previous studies, possibly due to the delay in the diagnosis in our patients. PMID- 16252196 TI - Investigation of fecal pancreatic elastase-1 levels in type 2 diabetic patients. AB - BACKGROUND/AIMS: Due to the close anatomical position between the endocrine system cells and the exocrine system cells in the pancreas, some interactions could be expected in these two different types of cells. This possible exocrine dysfunction may cause difficulties in the management of blood glucose level because of secondary malabsorption which may have resulted from the exocrine dysfunction. Taking this possibility into account, we aimed to investigate the exocrine function of the pancreas in 32 diabetic patients and in 12 healthy control subjects in this study. METHODS: Fecal pancreatic elastase-1 (PE1), which has a high sensitivity and specificity, was measured in serum samples by ELISA specifically for this purpose. RESULTS: It was found that the exocrine function declined in 28% of type 2 diabetic patients, while there was no decrease in the control subjects. However, there were no significant correlations between pancreatic elastase levels and the duration of diabetes, glycemic control, or consumption of alcohol. CONCLUSIONS: These findings suggest that evaluation of the exocrine function in diabetic patients might be useful for better management of diabetic patients. PMID- 16252197 TI - The value of serum zinc, copper, ceruloplasmin levels in patients with gastrointestinal tract cancers. AB - BACKGROUND/AIMS: Gastrointestinal cancers are frequent diseases of particular importance for surgeons with regard to their management and follow-up. Neoplastic diseases activate antioxidant defense systems. As a result, concentrations of redoxal enzymes and their co-factor elements appear to change. The levels of zinc (Zn), copper (Cu) and ceruloplasmin have been found to be critical parameters. In addition, it seems to be important to consider the Cu/Zn ratio in such cases. In this study, changes in the levels of Cu, Zn and ceruloplasmin in patients with gastrointestinal system cancers was evaluated, and the proportion of Cu to Zn (Cu/Zn) is discussed. METHODS: In this clinical trial, levels of Cu and Zn were defined with calorimetric methods and ceruloplasmin levels were measured with immunohistochemical methods in a control group of 20 healthy individuals and in 25 patients who underwent surgery for gastrointestinal system cancer. The blood samples were taken preoperatively in gastrointestinal system cancer patients. RESULTS: This study has shown that while the decrease in Zn levels and the increase in ceruloplasmin levels in patients with gastrointestinal system cancers were found to be significant (p<0. 001, p=0. 014) when compared to the control group, the increase in the level of Cu was also found to be significant (p=0. 019). In the patient group, the correlations between serum Cu and serum ceruloplasmin proved to be significant (r=991, p<0. 001). The Cu/Zn ratio, when compared with Zn, ceruloplasmin and Cu, showed significant results (r= 0. 562, r= 0. 500, r=0. 506, p<0. 001). CONCLUSIONS: This study shows that serum Cu, Zn and ceruloplasmin levels show changes in gastrointestinal system cancers, but further research is needed to demonstrate the importance and significance of these parameters and their relation with other contributing neoplastic factors. PMID- 16252198 TI - Effects of Momordica charantia L. (Cucurbitaceae) on indomethacin-induced ulcer model in rats. AB - BACKGROUND/AIMS: Fruits of Momordica charantia L.-cucurbitaceae have been frequently used in folk medicine for rapid healing of cutaneous lesions and peptic ulcer, especially in Western Anatolia in Turkey. METHODS: The anti ulcerogenic effect of the oily extract of Momordica charantia fruits was investigated in male Sprague-Dawley rats. Animals were separated into six groups. Distilled water (control group), famotidine (40 mg/kg), oily extracts (5 and 10 ml/kg), and vehicles (olive oil -5 and 10 ml/kg) were given orally (gavage). Thirty minutes later indomethacin (25 mg/kg) was administrated to all the groups. Six hours later, animals were killed with decapitation. For each stomach, ulcerated and total areas were measured (mm2). The ulcer indexes for each stomach and the ulcer inhibition rates for each group were calculated, after which the stomachs were evaluated pathologically (polymorphonuclear leukocytes infiltration). RESULTS: Ulcer inhibition rates were as follows: famotidine 91.54%, oily extract (5 ml/kg) -53.80%, oily extract (10 ml/kg) -98.04%, vehicle (olive oil -5 ml/kg) -18.40%, and vehicle (olive oil -10 ml/kg) -88.02%. According to polymorphonuclear leukocytes infiltration, oily extract (10 ml/kg) and vehicle (10 ml/kg) had similar effects to famotidine. CONCLUSIONS: The olive oil extract of M. charantia fruit did show a protective effect macroscopically. PMID- 16252199 TI - Turkish patients with osteoarthritis: their awareness of the side effects of NSAIDs. AB - BACKGROUND/AIMS: The study aim was to determine the awareness of Turkish osteoarthritis patients of the side effects of non-steroidal anti-inflammatory drugs. METHODS: The patients were interviewed by 138 doctors regarding the level of their knowledge of the side effects of non-steroidal anti-inflammatory drugs. RESULTS: A total of 3,755 patients (female/male: 3/1, 35% > 65 years) were included in the study. 35.5% of the patients were aware of side effects of non steroidal anti-inflammatory drugs. 85.4% and 11.5% were aware of the gastrointestinal and other system-related side effects, respectively. 51% had learned of the side effects from doctors, 19.8% received information from the package inserts, 21.3% had experienced side effects, and 10.0% and 0.8% had learned from their friends and pharmacist, respectively. CONCLUSIONS: Turkish osteoarthritis patients have a moderate level of knowledge of side effects of non steroidal anti-inflammatory drugs. Defining factors for knowledge of side effects of non-steroidal anti-inflammatory drugs were geographical region, socio-economic level and gender. This study reveals the physician's responsibility to educate patients about the side effects of non-steroidal anti-inflammatory drugs. PMID- 16252200 TI - Chronic intestinal pseudoobstruction: report of four pediatric patients. AB - Chronic intestinal pseudoobstruction is a rare disorder of intestinal motility, characterized by recurrence of continuous symptoms and signs of intestinal obstruction in the absence of true mechanical obstruction. Congenital or systemic disorders are the causes of chronic intestinal pseudoobstruction. The term idiopathic is applied when there is no congenital or secondary cause. Early diagnosis of intestinal pseudoobstruction is important to avoid repeated laparotomies. Treatment of chronic intestinal pseudoobstruction is usually supportive. Besides the supportive therapy, prokinetic agents such as erythromycin and octreotide are used in the therapy. In this article, four pediatric patients diagnosed as chronic intestinal pseudoobstruction are discussed with their clinical findings and laboratory abnormalities. The etiology of chronic intestinal pseudoobstruction was visceral myopathy in one patient. Two had idiopathic chronic intestinal pseudoobstruction and the other patient developed chronic intestinal pseudoobstruction after cardiac surgery. Erythromycin was administered to all four patients, one of whom did not respond to this therapy. Octreotide was effective in this case. PMID- 16252201 TI - MRCP findings of biliary ascariasis before and after medical treatment. AB - A 53-year-old woman presented with the complaints of abdominal pain on right upper quadrant, nausea, vomiting, diarrhea, and loss of appetite. Her physical exam was unremarkable except for right upper quadrant tenderness. Abdominal ultrasound revealed a tubular, mobile, non-shadowing echogenic structure within the slightly dilated common bile duct. Examination of feces for ova suggested the diagnosis of ascariasis. In this report, ultrasonographic and magnetic resonance cholangiopancreatographic findings of the case are described before and after medical treatment. PMID- 16252202 TI - The coexistence of hepatitis A and infectious mononucleosis. AB - Hepatitis A is an acute, self-limited disease that spreads predominantly by the fecal-oral route. Hepatitis A characteristically has an acute, sudden influenza like onset with a prominence of myalgia, headache, fever and malaise. Infectious mononucleosis is an acute illness characterized clinically by sore throat, fever and lymphadenopathy. The virus usually spreads from person to person by close contact with nasopharyngeal secretions. In this case the coexistence of both diseases in the same patient is found interesting. PMID- 16252203 TI - Esophageal tuberculosis mimicking esophageal carcinoma. AB - Esophageal tuberculosis is rare. In some cases, the clinical presentation of this infection may mimic esophageal carcinoma. Differential diagnosis may be difficult and may result in an unnecessary surgical therapy such as esophagectomy. In this report we document the endoscopical, radiological, histological and bacteriological features of esophageal tuberculosis in a 62-year-old woman. She was admitted to our hospital complaining of dysphagia and odynophagia. Upper gastrointestinal endoscopy revealed an ulcerovegetant lesion in the right wall of the esophagus suggesting esophageal carcinoma. Further investigation resulted in a diagnosis of esophageal tuberculosis. She was successfully treated by antituberculous chemotherapy. We suggest that esophageal tuberculosis has to be kept in mind in the differential diagnosis of esophageal ulcerovegetant lesions. PMID- 16252204 TI - A new and safe technique for removing cervical esophageal foreign body. AB - Despite the recent advances in instrumentation and anesthesia, removal of esophageal foreign bodies remains a challenge. Endoscopic removal of foreign bodies has yielded a success rate of 80%. Surgical removal of these foreign bodies is necessary when the endoscopic manipulations fail. Localization and size of the bodies play a critical role in the method of treatment. Here we present a patient with a large stone with sharp edges located in the cervico-thoracic region which was removed after being pushed into the hypopharynx through the esophagus rather than being pushed into the stomach. The technique used proved to be effective and safe; this may be the first use of the procedure in the literature. PMID- 16252205 TI - Common variable immunodeficiency (CVID) presenting with malabsorption due to giardiasis. AB - Common variable immunodeficiency is characterized with B-cell and T-cell dysfunction and hypogammaglobulinemia. Recurrent bacterial infections, such as otitis media, chronic sinusitis and recurrent pneumonia due to diminished immunoglobulin (Ig) levels and impaired antibody production are frequently observed in common variable immunodeficiency. Almost half of the patients with common variable immunodeficiency have problems related to the gastrointestinal system. A 39-year-old woman was referred to our department with the complaint of chronic diarrhea. She had experienced diarrhea without mucus or blood in the last year and had lost 30 kg. In her medical history, she had suffered from recurrent upper and lower respiratory infections like sinusitis, otitis media and pneumonia since childhood. Serum immunoglobulin levels were low. There were no parasites or ova in her stool examinations. Esophagogastroduodenoscopy detected widespread macroscopic nodular appearance on duodenum, and biopsies from the duodenum revealed giardiasis invading the tissue. She was diagnosed as common variable immunodeficiency. After metronidazole therapy and intravenous immunoglobulin infusion was started, her diarrhea attacks ceased and she regained her normal weight. Common gastrointestinal system problems in patients with common variable immunodeficiency are lactose intolerance, lymphoid hyperplasia/diffuse lymphoid infiltration, loss of villi and infection, especially with Giardia lamblia. Giardiasis may lead to severe mucosal flattening and sometimes to lymphoid hyperplasia at the lamina propria of the duodenum. Medical history should be evaluated carefully regarding recurrent respiratory infections. In such cases with chronic diarrhea, common variable immunodeficiency should be kept in mind as a possible cause. PMID- 16252206 TI - Morgagni hernia in an adult: atypical presentation and diagnostic difficulties. AB - Foramen of Morgagni hernias are rare diaphragmatic hernias, usually occurring on the right and located in the anterior mediastinum. Adult patients diagnosed with a foramen of Morgagni hernia are usually asymptomatic and associated with obesity, trauma or other causes of increased intraabdominal pressure. Plain pulmonary roentgenogram, radiological studies of the gastrointestinal system with contrast material, computerized tomography and magnetic resonance imaging studies are helpful in diagnosis. In this article, a 78-year-old female case with a Morgagni hernia incidentally diagnosed on chest X-ray is presented. PMID- 16252207 TI - Tuberculous peritonitis and elevated serum CA 125 in a patient with chronic renal failure. PMID- 16252208 TI - [Problems of emergency transfers of patients after a stroke. Results of a telemedicine pilot project for integrated stroke accommodation in southeast Bavaria (TEMPiS)]. AB - BACKGROUND: Specific stroke subtypes like subarachnoid hemorrhages or malignant brain infarcts require immediate interventions, but treatment options are offered mainly in specialized centers. For this reason, interhospital transfers from primary hospitals need to be done without delay. METHODS: The telemedic pilot project for integrative stroke care (TEMPiS) connects 2 stroke centers and 12 regional hospitals in Bavaria (Germany). Core elements are the implementation of stroke wards, telemedic consultation and improvement of emergency interhospital transfers. Organization of patient transports is offered by the central telemedic service. During the first 12 months of the continuing project all interhospital transfers initiated by the central telemedic service were prospectively documented. Emergency transports were analysed according to diagnosis, type of transport, distance and time delays. RESULTS: A total of 252 interhospital transfers were recommended in teleconsultations; finally 221 transports took place. Median total duration of transfers (including the necessary arrangements) was 134 min (interquartile range: 105-219) for intracerebral hemorrhages (N = 58), 138 min (95-157) for subarachnoid hemorrhages (N = 31), 161 min (100-230) for malignant infarcts (N = 22) and 147 min (109-180) for suspected basilar artery occlusion (N = 28). Time from admission in the primary hospital to initiation of interhospital transfer was 135 min (median; interquartile range: 86 172), transport time was 81 min (60-116). Helicopter transport did not save time for transfer distances up to 50 kilometres, compared to transport via ambulance (including assistance of hospital physicians). Transport using a special intensive care vehicle was much more time consuming because of the longer transport preparation time. CONCLUSION: Emergency transfers of stroke patients are time consuming. This may contribute to additional harm being done to severely ill patients. Faster organization and conduct of transports is required. PMID- 16252209 TI - [Permanent hypoparathyroidism due to thyroid cancer surgical procedures in patients exposed to radiation in the Chernobyl, Ukraine, nuclear reactor accident]. AB - BACKGROUND AND OBJECTIVE: Postoperative hypoparathyroidism is a rare, but complex endocrine disorder. The purpose of this case series study was to evaluate the symptoms, signs and sequelae of the disease in a major homogeneous patient group after the Chernobyl accident in the Ukraine. PATIENTS AND METHODS: This study is based on a random sample of 25 patients with symptomatic permanent hypoparathyroidism after surgical procedures on the thyroid for carcinoma following the Chernobyl accident in May 1986. They underwent physical examination and their parathyroid hormone and calcium levels in serum were determined. The signs, symptoms and sequelae of their disease were systematically assessed by a validated specific questionnaire. The extent of individual distress by the symptoms and signs were analysed by using a Likert scale (points between 1 = no and 5 = maximal distress). RESULTS: In 14 patients (56%) (age at time of operation: 18.24 +/- 4.26 years) paresthesias occurred at least every third day. Very frequent general symptoms were joint pains and hair loss (in 17 patients / 68%). 19 patients (76%) had enamel defects, 17 (68%) increasingly dry skin since the operation. Psychical symptoms like excitability (20 patients / 80%) and depressed mood since the operation (16 patients / 64%) were strikingly frequent. The patients were affected more by dry skin (2,76 on the Likert scale) than by paraesthesias (2,56) or joint pains (2,68) and most of all by depressive mood (2.96) and excitability (3.38). 4 patients (16%) had kidney stones, while osteoporosis and osteosclerosis occurred in 7 (28%) respectively 3 persons (12%). 6 patients (24%) had cataracts and 3 (12%) had basal ganglia calcification. Interestingly, 1/3 of all patients had only non-characteristic symptoms of the disease consistent with a latent form of hypoparathyroidism. CONCLUSIONS: Our data partly show a severe form of hypoparathyroidism of very young persons in the Chernobyl region, a finding which strongly supports the need of exact diagnosis and interdisciplinary treatment options of this postoperative disorder. PMID- 16252210 TI - [A case of autochthonous early summer meningoencephalitis in Mecklenberg-Western Pomerania]. AB - HISTORY: A 61-year-old man was bitten by a tick at Lake Woblitz, near the town of Neustrelitz in former East Germany. Nine days later he saw his general practitioner because of fever and headache. Three weeks after the tick bite he was hospitalized with fever (39.2 degrees C) and mental confusion. Because he had taken a Nile cruise six months earlier, malaria was considered and he was transferred to the department of tropical medicine and infectious diseases of the University of Rostock. INVESTIGATIONS: The patient was somnolent, his speech was slurred, and he had amnesic aphasia, as well as impaired fine motor control, but no meningism, focal signs, pyramidal tract or sensory impairment. Cerebrospinal fluid (CSF) showed mild lymphocytosis (9,400 leukocytes per microL; 89% lymphocytes) and elevated protein concentration (1322 mg/L) with blood brain barrier impairment and intrathecal IgM synthesis. Anti-tick-bite encephalitis (TBE) antibodies (ELISA: IgG and IgM) were present in serum and CSF, and serum immunofluorescence showed an eight-fold titer increase within two weeks. These findings confirm the diagnosis of TBE. Other infections (including those with cross-reacting flaviviruses) were excluded by appropriate antibody testing. THERAPY AND CLINICAL COURSE: There is no specific antiviral treatment for TBE, but on symptomatic therapy the patient recovered fully within four weeks. CONCLUSION: The site of the patient's infection is located 10 km to the west of an old TBE focus, but no TBE virus had been detected there after 1975. The case demonstrates that TBE should be included in the differential diagnosis of meningoencephalitis, even if the patient has not been in an acknowledged TBE endemic area. PMID- 16252212 TI - [Treatment of patients with carotid stenosis]. PMID- 16252213 TI - [Lipoprotein(a) in coronary heart disease: clinical significance and therapeutic options]. PMID- 16252214 TI - [The Emperor's New Clothes or: new structures for university hospitals]. PMID- 16252215 TI - [What is the "number at risk?"]. PMID- 16252216 TI - [See me. Feel me. Touch me? Heal me!]. PMID- 16252217 TI - [Efficacy of clonidine in paediatric anaesthesia]. AB - Clonidine is a mixed alpha (2)-/alpha (1)- adrenoceptor agonist. It decreases sympathetic tone and increases parasympathetic tone, which results in a lowering of blood pressure and heart rate. It was introduced as an antihypertensive medication in the late sixties. Yet Clonidine also has a sedative effect due to alpha (2)-adrenoceptor stimulation in the locus coeruleus and antinociceptive action caused through postjunctional noradrenergic pathways in the brainstem and spinal cord. This analgesic property of Clonidine has raised broad interest in its beneficial use in anaesthesiology. In adults and pediatric anaesthesia, clonidine can be safely and effectively used for premedication, intraoperative and postoperative analgesia, postoperative shivering, and for analgo-sedation in intensive care. This article will consider possible indications of Clonidine in children's anaesthesia and intensive care medicine. PMID- 16252218 TI - [Carotid-surgery in ultrasound-guided anesthesia of the regio colli lateralis]. AB - OBJECTIVE: Carotid endarterectomy significantly reduces the risk of stroke in patients with symptomatic and asymptomatic carotid artery stenosis. An increasing number of interventions in carotid surgery are performed under regional anesthesia in conscious patients. Carotid endarterectomy in local anesthesia requires block of the cervical nerves C2-C4, which may be accomplished in different ways. The most frequent method of regional anesthesia in carotid surgery is a combined block of profunda and superficial cervical plexus (by using a nerve stimulator). Ultrasound is frequently used in anesthesia for venous access and peripheral nerve block. By ultrasound, it is possible to visualize puncture needle and spread of local anesthetics at the correct position for block of cervical plexus in carotid surgery. The aim of the case reports was to demonstrate the effectiveness of using ultrasound for regional anesthesia in carotid surgery. METHOD: At the level of carotid bifurcation, scalene muscles were visualized by ultrasound (10-MHz-transducer). At this position, a puncture needle was inserted in the beam of ultrasound. Between the anterior scalene and the sternocleidomastoid muscles the local anesthetics were injected (10 ml prilocaine 1 %, 20 ropivacaine 0.375 %). In addition, superficial cervical plexus was performed by subcutaneous injection (10 ml prilocaine 1 %, 40 ropivacaine 0.375 %). Sedation was performed by application of remifentanil (max. 0.02 microg/kg/min), as needed. RESULTS: Ultrasound guided cervical block was performed in 29 patients. In all cases cervical plexus block was complete. A conversion to general anesthesia during operation was not necessary in any case. CONCLUSION: Ultrasound presents an alternative to cervical block using nerve stimulation, especially in patients with anatomical abnormalities. The method proves to be effective and may improve the approach to profundeal cervical plexus. PMID- 16252219 TI - [Influence of continuous positive airway pressure (CPAP) on blood flow velocity in the venous vascular system of the liver -- duplex sonographic investigations]. AB - OBJECTIVE: In the last years, noninvasive techniques of artificial ventilation like continuous positive airway pressure (CPAP) have become more and more important. However, the effects of such procedures on the circulation of the hepatosplanchnic area are, except for the indocyanine green plasma clearance, not yet representative by means of noninvasive investigation methods. METHODS: We studied maximum blood flow velocity in defined areas of hepatic and portal veins by duplex ultrasonography in 20 healthy volunteers (12 male, 8 female; age: 24,7 +/- 4,7 years [mean +/- SD]) before, during and after application of CPAP (10 cm H(2)O). As many measurements as possible were taken in both vascular systems. After division into five minute intervals arithmetical mean values were calculated for each interval. The experiment was divided into a control interval ("before CPAP"), three "CPAP" intervals and finally two "after CPAP" intervals. Heart rate and blood pressure were taken at the beginning of these intervals. RESULTS: Compared with the control interval, maximum blood flow velocity in the area of the hepatic vein decreased significantly by 13,2 %, 16,8 % and 15,7 % (p < or = 0,001 in each case) during each of the three "CPAP" periods. In the portal venous area blood flow velocity decreased by 4,3 %, 8,8 % and 7,0 %, respectively, with a significant decrease only in the second "CPAP" interval (p < or = 0,005). After CPAP, changes returned to baseline within a few minutes. Heart rate and blood pressure showed no significant differences during the whole experiment. CONCLUSION: Parameters taken in both venous vascular areas of the liver show clear correspondence with the results of former studies, which have been determined by means of more invasive investigation methods. Maximum blood flow velocity seems to be useful for monitoring liver blood flow and its individual time course during CPAP, noninvasively and semiquantitatively. Further trials under conditions of controlled mechanical ventilation should be performed to optimize ventilation strategies for the critically ill. PMID- 16252220 TI - [One-sided acquired pulmonary lobar emphysema -- successful treatment with selective intubation and one-sided high-frequency ventilation]. AB - Mechanical ventilation of preterm and newborn infants is associated with the risk of inducing a pulmonary baro- or volutrauma which may lead to one-sided acquired pulmonary lobar emphysema (APLE). We report a case of a preterm infant with severe APLE compromising hemodynamics due to mediastinal-shift, which was successfully treated with selective one-sided intubation and high-frequency ventilation. We assume that a brief malposition of the endotracheal tube initiated the emphysema of the left pulmonary lobe. We deduce recommendations for avoiding APLE from literature and our experience. PMID- 16252221 TI - [IXth Interdisciplinary CPA Symposium: Optimising the Oxygen Supply (Part I)]. PMID- 16252222 TI - [The optimization of oxygen levels]. PMID- 16252223 TI - [At Which Level is the PEEP Adequate in ARDS?]. PMID- 16252224 TI - [Optimization of oxygen delivery by mechanical ventilation -- new concepts]. PMID- 16252225 TI - [High FIO(2) during mechanical ventilation: pro -- con]. PMID- 16252226 TI - [Outcome improvement through FIO(2) 1.0: reality or myth?]. PMID- 16252227 TI - [Notes on the physiology and pathophysiology of cardiac output]. PMID- 16252228 TI - [Physiology and clinical relevance of hyperoxygenation]. PMID- 16252231 TI - High-resolution whole-genome association study of Parkinson disease. AB - We performed a two-tiered, whole-genome association study of Parkinson disease (PD). For tier 1, we individually genotyped 198,345 uniformly spaced and informative single-nucleotide polymorphisms (SNPs) in 443 sibling pairs discordant for PD. For tier 2a, we individually genotyped 1,793 PD-associated SNPs (P<.01 in tier 1) and 300 genomic control SNPs in 332 matched case-unrelated control pairs. We identified 11 SNPs that were associated with PD (P<.01) in both tier 1 and tier 2 samples and had the same direction of effect. For these SNPs, we combined data from the case-unaffected sibling pair (tier 1) and case unrelated control pair (tier 2) samples and employed a liberalization of the sibling transmission/disequilibrium test to calculate odds ratios, 95% confidence intervals, and P values. A SNP within the semaphorin 5A gene (SEMA5A) had the lowest combined P value (P=7.62 x 10(-6)). The protein encoded by this gene plays an important role in neurogenesis and in neuronal apoptosis, which is consistent with existing hypotheses regarding PD pathogenesis. A second SNP tagged the PARK11 late-onset PD susceptibility locus (P=1.70 x 10(-5)). In tier 2b, we also selected for genotyping additional SNPs that were borderline significant (P<.05) in tier 1 but that tested a priori biological and genetic hypotheses regarding susceptibility to PD (n=941 SNPs). In analysis of the combined tier 1 and tier 2b data, the two SNPs with the lowest P values (P=9.07 x 10(-6); P=2.96 x 10(-5)) tagged the PARK10 late-onset PD susceptibility locus. Independent replication across populations will clarify the role of the genomic loci tagged by these SNPs in conferring PD susceptibility. PMID- 16252233 TI - High-resolution identification of chromosomal abnormalities using oligonucleotide arrays containing 116,204 SNPs. AB - Mutation of the human genome ranges from single base-pair changes to whole chromosome aneuploidy. Karyotyping, fluorescence in situ hybridization, and comparative genome hybridization are currently used to detect chromosome abnormalities of clinical significance. These methods, although powerful, suffer from limitations in speed, ease of use, and resolution, and they do not detect copy-neutral chromosomal aberrations--for example, uniparental disomy (UPD). We have developed a high-throughput approach for assessment of DNA copy-number changes, through use of high-density synthetic oligonucleotide arrays containing 116,204 single-nucleotide polymorphisms, spaced at an average distance of 23.6 kb across the genome. Using this approach, we analyzed samples that failed conventional karyotypic analysis, and we detected amplifications and deletions across a wide range of sizes (1.3-145.9 Mb), identified chromosomes containing anonymous chromatin, and used genotype data to determine the molecular origin of two cases of UPD. Furthermore, our data provided independent confirmation for a case that had been misinterpreted by karyotype analysis. The high resolution of our approach provides more-precise breakpoint mapping, which allows subtle phenotypic heterogeneity to be distinguished at a molecular level. The accurate genotype information provided on these arrays enables the identification of copy neutral loss-of-heterozygosity events, and the minimal requirement of DNA (250 ng per array) allows rapid analysis of samples without the need for cell culture. This technology overcomes many limitations currently encountered in routine clinical diagnostic laboratories tasked with accurate and rapid diagnosis of chromosomal abnormalities. PMID- 16252232 TI - Mutations in TCF8 cause posterior polymorphous corneal dystrophy and ectopic expression of COL4A3 by corneal endothelial cells. AB - Posterior polymorphous corneal dystrophy (PPCD, also known as PPMD) is a rare disease involving metaplasia and overgrowth of corneal endothelial cells. In patients with PPCD, these cells manifest in an epithelial morphology and gene expression pattern, produce an aberrant basement membrane, and, sometimes, spread over the iris and nearby structures in a way that increases the risk for glaucoma. We previously mapped PPCD to a region (PPCD3) on chromosome 10 containing the gene that encodes the two-handed zinc-finger homeodomain transcription factor TCF8. Here, we report a heterozygous frameshift mutation in TCF8 that segregates with PPCD in the family used to map PPCD3 and four different heterozygous nonsense and frameshift mutations in TCF8 in four other PPCD probands. Family reports of inguinal hernia, hydrocele, and possible bone anomalies in affected individuals suggest that individuals with TCF8 mutations should be examined for nonocular anomalies. We detect transcripts of all three identified PPCD genes (VSX1, COL8A2, and TCF8) in the cornea. We show presence of a complex (core plus secondary) binding site for TCF8 in the promoter of Alport syndrome gene COL4A3, which encodes collagen type IV alpha 3, and we present immunohistochemical evidence of ectopic expression of COL4A3 in corneal endothelium of the proband of the original PPCD3 family. Identification of TCF8 as the PPCD3 gene provides a valuable tool for the study of critical gene regulation events in PPCD pathology and suggests a possible role for TCF8 mutations in altered structure and function of cells lining body cavities other than the anterior chamber of the eye. Thus, this study has identified TCF8 as the gene responsible for approximately half of the cases of PPCD, has implicated TCF8 mutations in developmental abnormalities outside the eye, and has presented the TCF8 regulatory target, COL4A3, as a key, shared molecular component of two different diseases, PPCD and Alport syndrome. PMID- 16252234 TI - Multiple correcting COL17A1 mutations in patients with revertant mosaicism of epidermolysis bullosa. AB - Revertant mosaicism by somatic reversion of inherited mutations has been described for a number of genetic diseases. Several mechanisms can underlie this reversion process, such as gene conversion, crossing-over, true back mutation, and second-site mutation. Here, we report the occurrence of multiple corrections in two unrelated probands with revertant mosaicism of non-Herlitz junctional epidermolysis bullosa, an autosomal recessive genodermatosis due to mutations in the COL17A1 gene. Immunofluorescence microscopy and laser dissection microscopy, followed by DNA and RNA analysis, were performed on skin biopsy specimens. In patient 1, a true back mutation, 3781T-->C, was identified in the specimen from the arm, and a second-site mutation, 4463-1G-->A, which compensated for the frameshift caused by the inherited 4424-5insC mutation, was identified in the 3' splice site of exon 55 in a specimen from the middle finger. Patient 2 showed- besides two distinct gene conversion events in specimens from the arm and hand sites, both of which corrected the 1706delA mutation--a second-site mutation (3782G-->C) in an ankle specimen, which prevented the premature ending of the protein by the 3781C-->T nonsense mutation (R1226X). Thus, both inherited mutations, paternal as well as maternal, reverted at least once by different reversion events in distinct cell clusters in the described patients. The occurrence of multiple correcting mutations within the same patient indicates that in vivo reversion is less unusual than was generally thought. Furthermore, in the male patient, mosaic patterns of type XVII collagen-positive keratinocytes were present in clinically unaffected and affected skin. This latter observation makes it likely that reversion may be overlooked and may happen more often than expected. PMID- 16252235 TI - Clinical and molecular findings in osteoporosis-pseudoglioma syndrome. AB - Mutations in the low-density lipoprotein receptor-related protein 5 gene (LRP5) cause autosomal recessive osteoporosis-pseudoglioma syndrome (OPPG). We sequenced the coding exons of LRP5 in 37 probands suspected of having OPPG on the basis of the co-occurrence of severe congenital or childhood-onset visual impairment with bone fragility or osteoporosis recognized by young adulthood. We found two putative mutant alleles in 26 probands, only one mutant allele in 4 probands, and no mutant alleles in 7 probands. Looking for digenic inheritance, we sequenced the genes encoding the functionally related receptor LRP6, an LRP5 coreceptor FZD4, and an LRP5 ligand, NDP, in the four probands with one mutant allele, and, looking for locus heterogeneity, we sequenced FZD4 and NDP in the seven probands with no mutations, but we found no additional mutations. When we compared clinical features between probands with and without LRP5 mutations, we found no difference in the severity of skeletal disease, prevalence of cognitive impairment, or family history of consanguinity. However, four of the seven probands without detectable mutations had eye pathology that differed from pathology previously described for OPPG. Since many LRP5 mutations are missense changes, to differentiate between a disease-causing mutation and a benign variant, we measured the ability of wild-type and mutant LRP5 to transduce Wnt and Norrin signal ex vivo. Each of the seven OPPG mutations tested, had reduced signal transduction compared with wild-type mutations. These results indicate that early bilateral vitreoretinal eye pathology coupled with skeletal fragility is a strong predictor of LRP5 mutation and that mutations in LRP5 cause OPPG by impairing Wnt and Norrin signal transduction. PMID- 16252237 TI - A constrained-likelihood approach to marker-trait association studies. AB - Marker-trait association analysis is an important statistical tool for detecting DNA variants responsible for genetic traits. In such analyses, an analysis model of the mean genetic effects of the genotypes is often specified. For instance, the effect of the disease allele on the trait is often specified to be dominant, recessive, additive, or multiplicative. Although this model-based approach is powerful when the analysis model is correctly specified, it has been found to have low power sometimes when the specified model is incorrect. We introduce an approach that does not require the specification of a particular genetic model. This approach is built upon a constrained maximum likelihood in which the mean genetic effect of the heterozygous genotype is required to not exceed those of the two homozygous genotypes. The asymptotic distribution of the likelihood-ratio statistic is derived for two special cases. A simulation study suggests that this new approach has power comparable to that of the model-based method when the analysis model is correctly specified. This approach uses one marker at a time (i.e., it is a single-marker analysis). However, given the latest findings that powerful inferential procedures for haplotype analyses can be constructed from single-marker analyses, we expect this approach to be useful for haplotype analyses. PMID- 16252236 TI - Handling marker-marker linkage disequilibrium: pedigree analysis with clustered markers. AB - Single-nucleotide polymorphisms (SNPs) are rapidly replacing microsatellites as the markers of choice for genetic linkage studies and many other studies of human pedigrees. Here, we describe an efficient approach for modeling linkage disequilibrium (LD) between markers during multipoint analysis of human pedigrees. Using a gene-counting algorithm suitable for pedigree data, our approach enables rapid estimation of allele and haplotype frequencies within clusters of tightly linked markers. In addition, with the use of a hidden Markov model, our approach allows for multipoint pedigree analysis with large numbers of SNP markers organized into clusters of markers in LD. Simulation results show that our approach resolves previously described biases in multipoint linkage analysis with SNPs that are in LD. An updated version of the freely available Merlin software package uses the approach described here to perform many common pedigree analyses, including haplotyping and haplotype frequency estimation, parametric and nonparametric multipoint linkage analysis of discrete traits, variance-components and regression-based analysis of quantitative traits, calculation of identity-by-descent or kinship coefficients, and case selection for follow-up association studies. To illustrate the possibilities, we examine a data set that provides evidence of linkage of psoriasis to chromosome 17. PMID- 16252238 TI - A novel method with improved power to detect recombination hotspots from polymorphism data reveals multiple hotspots in human genes. AB - We introduce a new method for detection of recombination hotspots from population genetic data. This method is based on (a) defining an (approximate) penalized likelihood for how recombination rate varies with physical position and (b) maximizing this penalized likelihood over possible sets of recombination hotspots. Simulation results suggest that this is a more powerful method for detection of hotspots than are existing methods. We apply the method to data from 89 genes sequenced in African American and European American populations. We find many genes with multiple hotspots, and some hotspots show evidence of being population-specific. Our results suggest that hotspots are randomly positioned within genes and could be as frequent as one per 30 kb. PMID- 16252239 TI - Severely incapacitating mutations in patients with extreme short stature identify RNA-processing endoribonuclease RMRP as an essential cell growth regulator. AB - The growth of an individual is deeply influenced by the regulation of cell growth and division, both of which also contribute to a wide variety of pathological conditions, including cancer, diabetes, and inflammation. To identify a major regulator of human growth, we performed positional cloning in an autosomal recessive type of profound short stature, anauxetic dysplasia. Homozygosity mapping led to the identification of novel mutations in the RMRP gene, which was previously known to cause two milder types of short stature with susceptibility to cancer, cartilage hair hypoplasia, and metaphyseal dysplasia without hypotrichosis. We show that different RMRP gene mutations lead to decreased cell growth by impairing ribosomal assembly and by altering cyclin-dependent cell cycle regulation. Clinical heterogeneity is explained by a correlation between the level and type of functional impairment in vitro and the severity of short stature or predisposition to cancer. Whereas the cartilage hair hypoplasia founder mutation affects both pathways intermediately, anauxetic dysplasia mutations do not affect B-cyclin messenger RNA (mRNA) levels but do severely incapacitate ribosomal assembly via defective endonucleolytic cleavage. Anauxetic dysplasia mutations thus lead to poor processing of ribosomal RNA while allowing normal mRNA processing and, therefore, genetically separate the different functions of RNase MRP. PMID- 16252240 TI - Promoter and 3'-untranslated-region haplotypes in the vitamin d receptor gene predispose to osteoporotic fracture: the rotterdam study. AB - Polymorphisms of the vitamin D receptor gene (VDR) have been shown to be associated with several complex diseases, including osteoporosis, but the mechanisms are unknown and study results have been inconsistent. We therefore determined sequence variation across the major relevant parts of VDR, including construction of linkage disequilibrium blocks and identification of haplotype alleles. We analyzed 15 haplotype-tagging SNPs in relation to 937 clinical fractures recorded in 6,148 elderly whites over a follow-up period of 7.4 years. Haplotype alleles of the 5' 1a/1e, 1b promoter region and of the 3' untranslated region (UTR) were strongly associated with increased fracture risk. For the 16% of subjects who had risk genotypes at both regions, their risk increased 48% for clinical fractures (P = .0002), independent of age, sex, height, weight, and bone mineral density. The population-attributable risk varied between 1% and 12% for each block and was 4% for the combined VDR risk genotypes. Functional analysis of the variants demonstrated 53% lower expression of a reporter construct with the 1e/1a promoter risk haplotype (P = 5 x 10(-7)) in two cell lines and 15% lower mRNA level of VDR expression constructs carrying 3'-UTR risk haplotype 1 in five cell lines (P = 2 x 10(-6)). In a further analysis, we showed 30% increased mRNA decay in an osteoblast cell line for the construct carrying the 3'-UTR risk haplotype (P = .02). This comprehensive candidate-gene analysis demonstrates that the risk allele of multiple VDR polymorphisms results in lower VDR mRNA levels. This could impact the vitamin D signaling efficiency and might contribute to the increased fracture risk we observed for these risk haplotype alleles. PMID- 16252241 TI - Evidence for widespread reticulate evolution within human duplicons. AB - Approximately 5% of the human genome consists of segmental duplications that can cause genomic mutations and may play a role in gene innovation. Reticulate evolutionary processes, such as unequal crossing-over and gene conversion, are known to occur within specific duplicon families, but the broader contribution of these processes to the evolution of human duplications remains poorly characterized. Here, we use phylogenetic profiling to analyze multiple alignments of 24 human duplicon families that span >8 Mb of DNA. Our results indicate that none of them are evolving independently, with all alignments showing sharp discontinuities in phylogenetic signal consistent with reticulation. To analyze these results in more detail, we have developed a quartet method that estimates the relative contribution of nucleotide substitution and reticulate processes to sequence evolution. Our data indicate that most of the duplications show a highly significant excess of sites consistent with reticulate evolution, compared with the number expected by nucleotide substitution alone, with 15 of 30 alignments showing a >20-fold excess over that expected. Using permutation tests, we also show that at least 5% of the total sequence shares 100% sequence identity because of reticulation, a figure that includes 74 independent tracts of perfect identity >2 kb in length. Furthermore, analysis of a subset of alignments indicates that the density of reticulation events is as high as 1 every 4 kb. These results indicate that phylogenetic relationships within recently duplicated human DNA can be rapidly disrupted by reticulate evolution. This finding has important implications for efforts to finish the human genome sequence, complicates comparative sequence analysis of duplicon families, and could profoundly influence the tempo of gene-family evolution. PMID- 16252242 TI - Curcumin treatment abrogates endoplasmic reticulum retention and aggregation induced apoptosis associated with neuropathy-causing myelin protein zero truncating mutants. AB - Mutations in MPZ, the gene encoding myelin protein zero (MPZ), the major protein constituent of peripheral myelin, can cause the adult-onset, inherited neuropathy Charcot-Marie-Tooth disease, as well as the more severe, childhood-onset Dejerine Sottas neuropathy and congenital hypomyelinating neuropathy. Most MPZ-truncating mutations associated with severe forms of peripheral neuropathy result in premature termination codons within the terminal or penultimate exons that are not subject to nonsense-mediated decay and are stably translated into mutant proteins with potential dominant-negative activity. However, some truncating mutations at the 3' end of MPZ escape the nonsense-mediated decay pathway and cause a mild peripheral neuropathy phenotype. We examined the functional properties of MPZ-truncating proteins that escaped nonsense-mediated decay, and we found that frameshift mutations associated with severe disease cause an intracellular accumulation of mutant proteins, primarily within the endoplasmic reticulum (ER), which induces apoptosis. Curcumin, a chemical compound derived from the curry spice tumeric, releases the ER-retained MPZ mutants into the cytoplasm accompanied by a lower number of apoptotic cells. Our findings suggest that curcumin treatment is sufficient to relieve the toxic effect of mutant aggregation-induced apoptosis and may potentially have a therapeutic role in treating selected forms of inherited peripheral neuropathies. PMID- 16252243 TI - Support for the homeobox transcription factor gene ENGRAILED 2 as an autism spectrum disorder susceptibility locus. AB - Our previous research involving 167 nuclear families from the Autism Genetic Resource Exchange (AGRE) demonstrated that two intronic SNPs, rs1861972 and rs1861973, in the homeodomain transcription factor gene ENGRAILED 2 (EN2) are significantly associated with autism spectrum disorder (ASD). In this study, significant replication of association for rs1861972 and rs1861973 is reported for two additional data sets: an independent set of 222 AGRE families (rs1861972 rs1861973 haplotype, P=.0016) and a separate sample of 129 National Institutes of Mental Health families (rs1861972-rs1861973 haplotype, P=.0431). Association analysis of the haplotype in the combined sample of both AGRE data sets (389 families) produced a P value of .0000033, whereas combining all three data sets (518 families) produced a P value of .00000035. Population-attributable risk calculations for the associated haplotype, performed using the entire sample of 518 families, determined that the risk allele contributes to as many as 40% of ASD cases in the general population. Linkage disequilibrium (LD) mapping with the use of polymorphisms distributed throughout the gene has shown that only intronic SNPs are in strong LD with rs1861972 and rs1861973. Resequencing and association analysis of all intronic SNPs have identified alleles associated with ASD, which makes them candidates for future functional analysis. Finally, to begin defining the function of EN2 during development, mouse En2 was ectopically expressed in cortical precursors. Fewer En2-transfected cells than controls displayed a differentiated phenotype. Together, these data provide further genetic evidence that EN2 might act as an ASD susceptibility locus, and they suggest that a risk allele that perturbs the spatial/temporal expression of EN2 could significantly alter normal brain development. PMID- 16252245 TI - Children with idiopathic hemihypertrophy and beckwith-wiedemann syndrome have different constitutional epigenotypes associated with wilms tumor. AB - Idiopathic hemihypertrophy (IH) is a congenital overgrowth syndrome associated with an increased risk of embryonal cancers in childhood. A related developmental disorder is Beckwith-Wiedemann syndrome (BWS), which increases risk for embryonal cancers, including Wilms tumor. Constitutional epigenetic alterations associated with BWS have been well characterized and include epigenetic alterations of imprinted genes on 11p15. The frequency of hypermethylation of H19 in children with IH and Wilms tumor, 20% (3/15), was significantly lower than the frequency in children with BWS and Wilms tumor, 79% (11/14; P = .0028). These results indicate that children with IH and Wilms tumor have different constitutional epigenotypes from those of children with BWS and Wilms tumor. PMID- 16252246 TI - Narrowing the candidate region for congenital diaphragmatic hernia in chromosome 15q26: contradictory results. PMID- 16252244 TI - The heritage of pathogen pressures and ancient demography in the human innate immunity CD209/CD209L region. AB - The innate immunity system constitutes the first line of host defense against pathogens. Two closely related innate immunity genes, CD209 and CD209L, are particularly interesting because they directly recognize a plethora of pathogens, including bacteria, viruses, and parasites. Both genes, which result from an ancient duplication, possess a neck region, made up of seven repeats of 23 amino acids each, known to play a major role in the pathogen-binding properties of these proteins. To explore the extent to which pathogens have exerted selective pressures on these innate immunity genes, we resequenced them in a group of samples from sub-Saharan Africa, Europe, and East Asia. Moreover, variation in the number of repeats of the neck region was defined in the entire Human Genome Diversity Panel for both genes. Our results, which are based on diversity levels, neutrality tests, population genetic distances, and neck-region length variation, provide genetic evidence that CD209 has been under a strong selective constraint that prevents accumulation of any amino acid changes, whereas CD209L variability has most likely been shaped by the action of balancing selection in non-African populations. In addition, our data point to the neck region as the functional target of such selective pressures: CD209 presents a constant size in the neck region populationwide, whereas CD209L presents an excess of length variation, particularly in non-African populations. An additional interesting observation came from the coalescent-based CD209 gene tree, whose binary topology and time depth (approximately 2.8 million years ago) are compatible with an ancestral population structure in Africa. Altogether, our study has revealed that even a short segment of the human genome can uncover an extraordinarily complex evolutionary history, including different pathogen pressures on host genes as well as traces of admixture among archaic hominid populations. PMID- 16252250 TI - Survey of the year 2004 commercial optical biosensor literature. AB - The year 2004 represents a milestone for the biosensor research community: in this year, over 1000 articles were published describing experiments performed using commercially available systems. The 1038 papers we found represent an approximately 10% increase over the past year and demonstrate that the implementation of biosensors continues to expand at a healthy pace. We evaluated the data presented in each paper and compiled a 'top 10' list. These 10 articles, which we recommend every biosensor user reads, describe well-performed kinetic, equilibrium and qualitative/screening studies, provide comparisons between binding parameters obtained from different biosensor users, as well as from biosensor- and solution-based interaction analyses, and summarize the cutting edge applications of the technology. We also re-iterate some of the experimental pitfalls that lead to sub-optimal data and over-interpreted results. We are hopeful that the biosensor community, by applying the hints we outline, will obtain data on a par with that presented in the 10 spotlighted articles. This will ensure that the scientific community at large can be confident in the data we report from optical biosensors. PMID- 16252251 TI - Association of the putative susceptibility gene, transient receptor potential protein melastatin type 2, with bipolar disorder. AB - Disturbed intracellular calcium (Ca(2+)) homeostasis has been implicated in bipolar disorder (BD). Reduced mRNA levels of the transient receptor potential Ca(2+) permeable channel melastatin type 2, TRPM2, in B lymphoblast cell lines (BLCL) from bipolar I disorder (BD-I) patients showing elevated basal intracellular Ca(2+) ([Ca(2+)](B)), an index of altered intracellular Ca(2+) homeostasis, along with its location within a putative BD susceptibility locus (21q22.3), implicates the involvement of this gene in the Ca(2+) abnormalities and the genetic diathesis to BD. We tested this hypothesis by examining the association of selected single nucleotide polymorphisms (SNPs) and their haplotypes, spanning the TRPM2 gene, with BD and BLCL [Ca(2+)](B), in a case control design. The 5' TaqMan SNP assay was used to detect selected SNPs. BLCL [Ca(2+)](B) was determined by ratiometric fluorometry. SNP rs1618355 in intron 18 was significantly associated with BD as a whole (P < 7.0 x 10(-5); odds ratio (OR) = 2.60), and when stratified into BD-I (P < 7.0 x 10(-5), OR = 2.48) and BD II (P = 7.0 x 10(-5), OR = 2.88) subgroups. In addition, the alleles of the individual SNPs forming a seven marker at-risk haplotype were in excess in BD (12.0% in BD vs. 0.9% in controls; P = 2.3 x 10(-12)). A weak relationship was also detected between BLCL [Ca(2+)](B) and TRPM2 SNP rs1612472 in intron 19. These findings suggest genetic variants of the TRPM2 gene increase risk for BD and support the notion that TRPM2 may be involved in the pathophysiology of BD. PMID- 16252253 TI - Interaction of ICAM-1 with beta 2-integrin CD11c/CD18: characterization of a peptide ligand that mimics a putative binding site on domain D4 of ICAM-1. AB - The integrin CD11c/CD18 plays a role in leukocyte and cell matrix adhesion and is highly expressed in certain hematopoietic malignancies. To better characterize ligand binding properties, we panned random peptide phage-display libraries over purified CD11c/CD18. We identified a phage expressing the circular peptide C GRWSGWPADL-C. C-GRWSGWPADL-C phage bound specifically to CD11c/CD18 expressing monocytes but not CD11c/CD18 negative lymphocytes and showed 5 x 10(3)-fold higher binding to purified CD11c/CD18 than control phage, without binding to CD11b/CD18. Peptide sequence analysis revealed a similar sequence in domain D5 of ICAM-1 and an alternative, phase-shifted motif in domain D4. Surface plasmon resonance experiments demonstrated direct interaction of ICAM-1 and CD11c/CD18. A soluble fusion protein containing the extracellular domain of ICAM-1 abolished C GRWSGWPADL-C phage binding to CD11c/CD18. Moreover, synthetic monomeric circular peptide C-GRWSGWPADL-C bound specifically to CD11c/CD18 and inhibited ICAM-1 binding. Its rather low binding affinity and inability to displace pentavalent C GRWSGWPADL-C phage from CD11c/CD18 suggests that a multimeric display of the selected peptide is essential for high affinity binding. Using ICAM-1 deletion constructs, we showed that domain D4 is required for interaction with CD11c/CD18, suggesting that C-GRWSGWPADL-C phage binds specifically to CD11c/CD18 by structurally mimicking the interaction site on D4 of ICAM-1. PMID- 16252254 TI - Dehydroepiandrosterone replacement in patients with Addison's disease has a bimodal effect on regulatory (CD4+CD25hi and CD4+FoxP3+) T cells. AB - Oral replacement of the near-total deficiency of dehydroepiandrosterone (DHEA) in patients with Addison's disease (adrenal insufficiency) enhances mood and well being and reduces fatigue. We studied the immunological effects of 12 wk of oral DHEA treatment in ten patients with Addison's disease receiving their normal mineralo- and glucocorticoid hormone replacement. We found that baseline circulating regulatory T cells were reduced in Addison's disease patients compared to controls, a hitherto unrecognised defect in this disorder. Oral DHEA treatment had a bimodal effect on naturally occurring regulatory (CD4+CD25hiFoxP3+) T cells and lymphocyte FoxP3 expression. Oral DHEA replacement restored normal levels of regulatory T cells and led to increased FoxP3 expression. These effects were probably responsible for a suppression of constitutive cytokine expression following DHEA withdrawal. In contrast, oral DHEA treatment led to reduced FoxP3 expression induced by TCR engagement and so augmented the cytokine response, but without a bias towards the Th1 or Th2 phenotype. NK and NKT cell numbers fell during DHEA treatment, and homeostatic lymphocyte proliferation was increased. We conclude that DHEA replacement in Addison's disease has significant immunomodulatory properties and propose that it has a greater impact on the human immune system than would be expected from its classification as a dietary supplement. PMID- 16252255 TI - Identification and determination of major flavonoids in rat urine by HPLC-UV and HPLC-MS methods following oral administration of Dalbergia odorifera extract. AB - Flavonoids are the main active constituents of Dalbergia odorifera. The excretion of the major flavonoids in rat urine after oral administration of D. odorifera extract was investigated by HPLC-UV and HPLC-MS methods. Utilizing the HPLC-MS technique, 18 flavonoids, including five isoflavones, four isoflavanones, four neoflavones, two flavanones, two chalcones and one isoflavanonol were identified in free form in a urine sample based on the direct comparison of the corresponding tR, UV maximum absorbance (lambda(max)) values and MS data with the authentic standards. The amounts of the prominent flavonoids, (3R)-4'-methoxy 2',3,7-trihydroxyisoflavanone and vestitone, were determined by HPLC-UV with the internal standard method, and the validation procedure confirmed that it afforded reliable analysis of these two analytes in urine after oral administration of D. odorifera extract. PMID- 16252256 TI - Attenuation of half sulfur mustard gas-induced acute lung injury in rats. AB - Airway instillation into rats of 2-chloroethyl ethyl sulfide (CEES), the half molecule of sulfur mustard compound, results in acute lung injury, as measured by the leak of plasma albumin into the lung. Morphologically, early changes in the lung include alveolar hemorrhage and fibrin deposition and the influx of neutrophils. Following lung contact with CEES, progressive accumulation of collagen occurred in the lung, followed by parenchymal collapse. The co instillation with CEES of liposomes containing pegylated (PEG)-catalase (CAT), PEG-superoxide dismutase (SOD), or the combination, greatly attenuated the development of lung injury. Likewise, the co-instillation of liposomes containing the reducing agents, N-acetylcysteine (NAC), glutathione (GSH), or resveratrol (RES), significantly reduced acute lung injury. The combination of complement depletion and airway instillation of liposomes containing anti-oxidant compounds maximally attenuated CEES-induced lung injury by nearly 80%. Delayed airway instillation of anti-oxidant-containing liposomes (containing NAC or GSH, or the combination) significantly diminished lung injury even when instillation was delayed as long as 1 h after lung exposure to CEES. These data indicate that CEES induced injury of rat lungs can be substantially diminished by the presence of reducing agents or anti-oxidant enzymes delivered via liposomes. PMID- 16252257 TI - Effects of interleukins 2 and 12 on TBT-induced alterations of MAP kinases p38 and p44/42 in human natural killer cells. AB - NK cells are lymphocytes in the non-adaptive immune system that protect the body against intracellular pathogens and eliminate tumor cells. Tributyltin (TBT) is a toxic chemical that has been detected in human foods as well as in human blood. The role of TBT in immunosuppression has been described, including inhibition of the human NK-cell cytotoxic function. Previous studies indicated that exposure of NK cells to TBT for 1 h induced progressive and irreversible inhibition of cytotoxic function. However, it was found that if NK cells were incubated in TBT free media with either IL-2 or IL-12, loss of cytotoxic function was prevented/reversed within 24 h. Molecular studies established that loss of cytotoxic function is accompanied by alteration of MAP kinases (MAPKs) p38 and p44/42 phosphorylation. This study examined whether interleukin-mediated recovery of cytotoxicity involved reversal of tributyltin-altered p38 and p44/42 phosphorylation. The results indicated that there was no substantial IL-2 prevention/reversal of the TBT-induced alteration of phosphorylation of either p38 or p44/42 after either a 24 or 48 h recovery period. Additionally, IL-12 caused no substantial prevention/reversal of the TBT-induced alteration of phosphorylation of the MAPKs seen after either 24 or 48 h. These data suggest that IL-2 and/or IL-12-mediated recovery of NK cytotoxic function is not a result of prevention/reversal of TBT-induced phosphorylation of p38 and p44/42 MAPKs at the 24 or 48 h time points. PMID- 16252258 TI - Toxicology of fluoroacetate: a review, with possible directions for therapy research. AB - Fluoroacetate (FA; CH2FCOOR) is highly toxic towards humans and other mammals through inhibition of the enzyme aconitase in the tricarboxylic acid cycle, caused by 'lethal synthesis' of an isomer of fluorocitrate (FC). FA is found in a range of plant species and their ingestion can cause the death of ruminant animals. Some fluorinated compounds -- used as anticancer agents, narcotic analgesics, pesticides or industrial chemicals -- metabolize to FA as intermediate products. The chemical characteristics of FA and the clinical signs of intoxication warrant the re-evaluation of the toxic danger of FA and renewed efforts in the search for effective therapeutic means. Antidotal therapy for FA intoxication has been aimed at preventing fluorocitrate synthesis and aconitase blockade in mitochondria, and at providing citrate outflow from this organelle. Despite a greatly improved understanding of the biochemical mechanism of FA toxicity, ethanol, if taken immediately after the poisoning, has been the most acceptable antidote for the past six decades. This review deals with the clinical signs and physiological and biochemical mechanisms of FA intoxication to provide an explanation of why, even after decades of investigation, has no effective therapy to FA intoxication been elaborated. An apparent lack of integrated toxicological studies is viewed as a limiter of progress in this regard. Two principal ways of developing effective therapies for FA intoxication are considered. Firstly, competitive inhibition of FA interaction with CoA and of FC interaction with aconitase. Secondly, channeling the alternative metabolic pathways by orienting the fate of citrate via cytosolic aconitase, and by maintaining the flux of reducing equivalents into the TCA cycle via glutamate dehydrogenase. PMID- 16252260 TI - Ferritin light chain down-modulation generates depigmentation in human metastatic melanoma cells by influencing tyrosinase maturation. AB - Recently, after the identification of ferritin light chain (L-ferritin) gene and protein over-expression in human metastatic melanoma cells, we engineered, starting from the LM metastatic melanoma cell line, clones in which L-ferritin gene expression was down-regulated by the stable expression of a specific antisense construct. The present investigation started from the observation that L-ferritin down-regulated LM cells displayed a less pigmented phenotype, confirmed by a major decrease of total melanin, when compared to control LM cells. This finding was accompanied by a dramatic decrease in tyrosinase activity, which was not paralleled by a concomitant reduction of the amount of tyrosinase specific mRNA. Western blot analysis of tyrosinase in control LM cells displayed a pattern, which corresponds to the progressive glycosylation of the native protein up to the 80 kDa form, considered the functional one. Tyrosinase pattern assayed in L-ferritin down-regulated LM cells showed the remarkable absence of the 80 kDa form and a prevalence of endoglycosidase H (endo H) sensitive immature (70 kDa) tyrosinase, accumulated in the endoplasmic reticulum (ER), as confirmed by confocal microscopy analysis. These results demonstrate that, in a human metastatic melanoma cell line, the stress condition promoted by L-ferritin down-modulation, can substantially influence proper maturation of tyrosinase. PMID- 16252259 TI - Phenotypic and functional changes of human melanoma xenografts induced by DNA hypomethylation: immunotherapeutic implications. AB - Emerging in vitro evidence points to an immunomodulatory activity of DNA hypomethylating drugs in human malignancies. We investigated the potential of 5 aza-2'-deoxycytidine (5-AZA-CdR) to modulate the expression of cancer testis antigens (CTA) and of HLA class I antigens by melanoma xenografts, and the resulting modifications in immunogenicity of neoplastic cells. Three primary cultures of melanoma cells, selected for immune phenotype and growth rate, were grafted into BALB/c nu/nu mice that were injected intraperitoneally with different dose- and time-schedules of 5-AZA-CdR. Molecular analyses demonstrated a de novo long-lasting expression of the CTA MAGE-1, -2, -3, -4, -10, GAGE 1-6, NY-ESO-1, and the upregulation of MAGE-1, MAGE-3, and NY-ESO-1 levels in melanoma xenografts from 5-AZA-CdR-treated mice. Serological and biochemical analyses identified a de novo expression of NY-ESO-1 protein and a concomitant and persistent upregulation of HLA class I antigens and of HLA-A1 and -A2 alleles. Immunization of BALB/c mice with 5-AZA-CdR-treated melanoma cells generated high titer circulating anti-NY-ESO-1 antibodies. Altogether, the data obtained identify an immunomodulatory activity of 5-AZA-CdR in vivo and strongly suggest for its clinical use to design novel strategies of CTA-based chemo-immunotherapy for melanoma patients. PMID- 16252261 TI - Androgen inducibility of Fgf8 in Shionogi carcinoma 115 cells correlates with an adjacent t(5;19) translocation. AB - Fgf8 (fibroblast growth factor 8) was initially cloned from a mouse mammary tumor cell line derived from the androgen-dependent Shionogi carcinoma 115. The androgen-inducible expression of Fgf8 in this tumor controls its androgen dependent phenotype, thus stimulating interest in this gene as a possible factor in human prostate cancer and other androgen-sensitive cancers. However, apart from Shionogi carcinoma 115, the androgen inducibility of Fgf8 is controversial. In the present study, having not detected androgen-inducible expression of Fgf8 in other mouse mammary cell lines or mouse prostate, we examined the Shionogi carcinoma 115-derived S115 cell line for mouse mammary tumor virus (MMTV) insertions or other nearby DNA rearrangements that might explain the androgen inducibility of Fgf8 in these cells. Southern blotting did not detect MMTV insertions near Fgf8 but did reveal a specific DNA rearrangement 3.7 kb upstream of Fgf8 in S115 cells and in other cells (SC115) independently derived from Shionogi carcinoma 115. Spectral karyotyping of S115 cells and sequencing of the cloned rearrangement junctions indicate that Fgf8 is involved in a t(5;19) translocation. The chromosome 5 sequence joined to Fgf8 is immediately adjacent to Smr2 (submaxillary gland androgen-regulated protein 2) and includes Muc10 (mucin 10), two genes that we show are testosterone inducible in S115 cells, suggesting that the androgen-dependent expression of Fgf8 in Shionogi carcinoma 115 and derivative cells results from this translocation. Together, these results suggest that androgen inducibility is not an inherent property of the Fgf8 gene, which has implications regarding this gene's proposed role in the etiology of hormone-responsive cancers. PMID- 16252262 TI - miR-155/BIC as an oncogenic microRNA. PMID- 16252263 TI - t(3;14)(p14;q32) results in aberrant expression of FOXP1 in a case of diffuse large B-cell lymphoma. AB - Strong expression of Forkhead box-P1 (FOXP1), a winged-helix transcription factor, has been identified as an independent prognostic factor in diffuse large B-cell lymphoma (DLBCL). However, possible mechanisms of deregulation of this gene, on 3p14.1, have yet to be elucidated. We have identified a breakpoint at the IGA1 gene in the immunoglobulin heavy chain (IGH) locus at 14q32 that was juxtaposed to the FOXP1 gene locus in a gastric DLBCL that showed strong expression of FOXP1. This may be one possible mechanism of deregulating FOXP1 expression by placing it under the control of IGH enhancers. PMID- 16252264 TI - Simultaneous determination of rifampicin and sulbactam in mouse plasma by high performance liquid chromatography. AB - A simple and rapid high-performance liquid chromatographic (HPLC) method with ultraviolet detection has been developed and validated for the simultaneous determination of rifampicin and sulbactam in mouse plasma. Plasma samples were deproteinized with acetonitrile and separated by HPLC on a RP-18 (125 x 4 mm, 5 microm) column and gradient elution with potassium dihydrogen phosphate solution (pH 4.5; 50 mm) and acetonitrile at a flow-rate of 1.0 mL/min. Rifampicin and sulbactam were monitored at 230 nm and confirmed by means of their UV spectra using a diode-array detector. The method was linear at plasma levels from 1 to 100 microg/mL for rifampicin and from 5 to 200 microg/mL for sulbactam. The limits of quantification were 0.6 microg/mL for rifampicin and 4.2 microg/mL for sulbactam. The intra- and inter-day precisions of the method (RSD) were lower than 5% for both compounds. Average recoveries of rifampicin and sulbactam from mice plasma were 98.2 and 89.3%, respectively. The developed method was successfully applied to the determination of the pharmacokinetic profile of both compounds in mice. PMID- 16252265 TI - Modelling age-dependent force of infection from prevalence data using fractional polynomials. AB - The force of infection is one of the primary epidemiological parameters of infectious diseases. For many infectious diseases it is assumed that the force of infection is age-dependent. Although the force of infection can be estimated directly from a follow up study, it is much more common to have cross-sectional seroprevalence data from which the prevalence and the force of infection can be estimated. In this paper, we propose to model the force of infection within the framework of fractional polynomials. We discuss several parametric examples from the literature and show that all of these examples can be expressed as special cases of fractional polynomial models. We illustrate the method on five seroprevalence samples, two of Hepatitis A, and one of Rubella, Mumps and Varicella. PMID- 16252266 TI - Molecular evolution of evolutionary novelties: the vagina and uterus of therian mammals. AB - Innovations are an integral part of the evolutionary process if we accept the fact that more complex organisms derived from anatomically simple ones. All major taxa are distinguished not only by their closer genealogical relatedness relative to other species but also by the possession of novel anatomical and physiological features. The question is whether the origin of these novel characters can be simply understood as adaptations, like all other phenotypic differences that arise by natural selection, or whether the origin of these characters requires more profound genetic changes. In this paper, we argue that innovations constitute a distinct class of evolutionary processes that require a research program complementary to the study of adaptation. The distinguishing feature of innovations is the origin of novel organ identity gene functions specific to the novel character. By implication, research into the origin of novel characters has to identify the developmental regulatory links that were involved in the evolution of these characters. We suggest that novel regulatory links will include the evolution of cis-regulatory elements as well as novel protein-protein interactions among transcription factor proteins. The latter hypothesis suggests that innovations should leave a trace in the evolution of the protein coding regions of transcription factor genes. We illustrate this idea with results on the evolution of HoxA-11 and HoxA-13 in the stem lineage of placental mammals. These genes are essential for female reproductive tract development and function. We show that, as predicted, these genes experience strong directional selection in the stem lineage of placental mammals and that these amino acid substitutions affect residues at the surface of the protein, consistent with their expected role in protein-protein interactions. We conclude that a careful analysis of sequence variation in developmental genes can aid in testing which developmental changes were instrumental in the origin of novel morphological characters. PMID- 16252267 TI - Editorial: evolutionary innovation and morphological novelty. PMID- 16252268 TI - Design of multi-centre trials with binary response. AB - We propose a correlated beta-binomial model for the binary response in multi centre trials. The likelihood function in this case has a closed-form and we avoid multivariate numerical integrations in determining the maximum likelihood estimator. Based on derived asymptotic variance-covariance matrix of the MLE, we obtain relatively simple formulae that relate the number of centres, the total number of patients and the precision of the parameter estimate. PMID- 16252269 TI - Confidence intervals for an effect size measure based on the Mann-Whitney statistic. Part 1: general issues and tail-area-based methods. AB - For two random variables X and Y, theta = Pr[Y > X] + (1/2)Pr[Y = X] is advocated as a general measure of effect size to characterize the degree of separation of their distributions. It is estimated by U/mn, a generalization of the Mann Whitney U statistic, derived by dividing U by the product of the two sample sizes. It is equivalent to the area under the receiver operating characteristic curve. It is readily visualized in terms of two Gaussian distributions with appropriately separated peaks. The effect of discretization of a continuous variable is explored. Tail-area-based confidence interval methods are developed which can be applied to very small samples or extreme outcomes. PMID- 16252270 TI - Bayesian analysis of the differences of count data. AB - Paired count data usually arise in medicine when before and after treatment measurements are considered. In the present paper we assume that the correlated paired count data follow a bivariate Poisson distribution in order to derive the distribution of their difference. The derived distribution is shown to be the same as the one derived for the difference of the independent Poisson variables, thus recasting interest on the distribution introduced by Skellam. Using this distribution we remove correlation, which naturally exists in paired data, and we improve the quality of our inference by using exact distributions instead of normal approximations. The zero-inflated version is considered to account for an excess of zero counts. Bayesian estimation and hypothesis testing for the models considered are discussed. An example from dental epidemiology is used to illustrate the proposed methodology. PMID- 16252271 TI - Correcting for selection using frailty models. AB - Chronic diseases are roughly speaking lifelong transitions between the states: relapse and recovery. The long-term pattern of recurrent times-to-relapse can be investigated with routine register data on hospital admissions. The relapses become readmissions to hospital, and the time spent in hospital are gaps between subsequent times-at-risk. However, problems of selection and dependent censoring arise because the calendar period of observation is limited and the study population likely to be heterogeneous. We will theoretically verify that an assumption of conditional independence of all times-at-risk and gaps, given the latent individual frailty level, allows for consistent inference in the shared frailty model. Using simulation studies, we also investigate cases where gaps (and/or staggered entry) are informative for the individual frailty. We found that the use of the shared frailty model can be extended to situations, where gaps are dependent on the frailty, but short compared to the distribution of the times-to-relapse. Our motivating example deals with the course of schizophrenia. We analysed routine register data on readmissions in almost 9000 persons with the disorder. Marginal survival curves of time-to-first-readmission, time-to-second readmission, etc. were estimated in the shared frailty model. Based on the schizophrenia literature, the conclusion of our analysis was rather surprising: one of a stable course of disorder. PMID- 16252272 TI - Statistical evaluation of biomarkers as surrogate endpoints: a literature review. AB - A valid surrogate endpoint allows correct inference to be drawn regarding the effect of an intervention on the unobserved true clinical endpoint of interest. The perceived practical and ethical advantages of substituting a surrogate endpoint for a clinical endpoint have led to a considerable number of statistical methods being proposed for the evaluation of a biomarker as a surrogate endpoint. We review the main statistical schools of thought which have developed and consider how the validation process might be arranged within the regulatory and practical constraints of the drug development process. We conclude by assessing which of the candidate statistical methods offer the best approach for surrogate endpoint evaluation. PMID- 16252274 TI - Multiple hypothesis testing strategies for genetic case-control association studies. AB - The genetic case-control association study of unrelated subjects is a leading method to identify single nucleotide polymorphisms (SNPs) and SNP haplotypes that modulate the risk of complex diseases. Association studies often genotype several SNPs in a number of candidate genes; we propose a two-stage approach to address the inherent statistical multiple comparisons problem. In the first stage, each gene's association with disease is summarized by a single p-value that controls a familywise error rate. In the second stage, summary p-values are adjusted for multiplicity using a false discovery rate (FDR) controlling procedure. For the first stage, we consider marginal and joint tests of SNPs and haplotypes within genes, and we construct an omnibus test that combines SNP and haplotype analysis. Simulation studies show that when disease susceptibility is conferred by a SNP, and all common SNPs in a gene are genotyped, marginal analysis of SNPs using the Simes test has similar or higher power than marginal or joint haplotype analysis. Conversely, haplotype analysis can be more powerful when disease susceptibility is conferred by a haplotype. The omnibus test tracks the more powerful of the two approaches, which is generally unknown. Multiple testing balances the desire for statistical power against the implicit costs of false positive results, which up to now appear to be common in the literature. PMID- 16252273 TI - Turner syndrome: four challenges across the lifespan. AB - Turner syndrome (TS) is a sex chromosome condition that occurs in approximately 1/2,500 live female births. Despite the prevalence of this chromosomal condition, the challenges these women face throughout their lives are not fully understood. This qualitative research study aimed to characterize the subjective experiences of individuals with TS throughout their lifespan, to investigate their concerns and obstacles, and to offer insight into the strengths and weaknesses of health care delivery, as they perceived them. Ninety-seven girls and women with TS and 21 parents consented to participate in this interview study. Interviews were semi structured and open-ended in design. Questions sought to elicit responses relating to existing concerns associated with their condition and positive and negative health care experiences. Participants were divided into four age categories (childhood, adolescence, adulthood, and mature adulthood) to facilitate a comparative analysis across the age spectrum. Regardless of age, infertility was the most frequently cited concern followed closely by short stature. Sexual development and function and general health were also viewed as challenges by a number of participants in each age group. Although the relative weight of these four concerns tended to shift based upon the individual's age and life experiences, all four issues remained significant throughout the lifespan. Enhanced awareness of the evolving physical and psychological challenges faced by girls and women with TS may help health care providers (HCPs) improve the quality of life for these individuals. PMID- 16252275 TI - A mixed effects Markov model for repeated binary outcomes with non-ignorable dropout. AB - In many areas of research, repeated binary measures often represent a two-state stochastic process, where individuals can transition among two states. In a behavioural or physical disability setting, individuals can flow from susceptible or subthreshold state, to an infectious or symptomatic state, and back to a subthreshold state. Quite often the transition among the states happens in continuous time but is observed at discrete, irregularly spaced timepoints which may be unique to each individual. Methods for analyses of such data are typically based on the Markov assumption. Cook (Biometrics 1999; 55:915-920) introduced a conditional Markov model that accommodates the subject-to-subject variation in the model parameters with random effects. We extend this model by adding a non ignorable dropout component to the model. Specification of the distribution of the random effects is made to guarantee a closed form expression of the marginal likelihood. This methodology is illustrated by applications to a data set from a parasitic field infection survey, a data set from a cocaine treatment study, and a data set from an aging study. Simulations suggest that the shared parameter model is robust with respect to at least one alternative non-ignorable model. PMID- 16252276 TI - Evolution and developmental patterning of the vertebrate skeletal muscles: perspectives from the lamprey. AB - The myotome in gnathostome vertebrates, which gives rise to the trunk skeletal muscles, consists of epaxial (dorsal) and hypaxial (ventral) portions, separated by the horizontal myoseptum. The hypaxial portion contains some highly derived musculature that is functionally as well as morphologically well differentiated in all the gnathostome species. In contrast, the trunk muscles of agnathan lampreys lack these distinctions and any semblance of limb muscles. Therefore, the lamprey myotomes probably represent a primitive condition compared with gnathostomes. In this review, we compare the patterns of expression of some muscle-specific genes between the lamprey and gnathostomes. Although the cellular and tissue morphology of lamprey myotomes seems uniform and undifferentiated, some of the muscle-specific genes are expressed in a spatially restricted manner. The lamprey Pax3/7 gene, a cognate of gnathostome Pax3, is expressed only at the lateral edge of the myotomes and in the hypobranchial muscle, which we presume is homologous to the gnathostome hypobranchial muscle. Thus, the emergence of some part of a hypaxial-specific gene regulatory cascade might have evolved before the agnathan/gnathostome divergence, or before the evolutionary separation of epaxial and hypaxial muscles. PMID- 16252277 TI - Bidirectional fusion of the heart-forming fields in the developing chick embryo. AB - It is generally thought that the early pre-tubular chick heart is formed by fusion of the anterior or cephalic limits of the paired cardiogenic fields. However, this study shows that the heart fields initially fuse at their midpoint to form a transitory "butterfly"-shaped, cardiogenic structure. Fusion then progresses bi-directionally along the longitudinal axis in both cranial and caudal directions. Using in vivo labeling, we demonstrate that cells along the ventral fusion line are highly motile, crossing future primitive segments. We found that mesoderm cells migrated cephalically from the unfused tips of the anterior/cephalic wings into the head mesenchyme in the region that has been called the secondary heart field. Perturbing the anterior/cranial fusion results in formation of a bi-conal heart. A theoretical role of the ventral fusion line acting as a "heart organizer" and its role in cardia bifida is discussed. PMID- 16252279 TI - Bulimia nervosa in an elderly male: a case report. AB - OBJECTIVE: There is increasing knowledge regarding demographics, comorbid psychiatric disorders, and symptoms characteristic of males suffering from eating disorders. METHOD: The current case report describes the history, symptom progression, and successful treatment of an elderly bulimic male whose presentation was atypical. RESULTS: This case underscores the need to consider the presence of an eating disorder regardless of age and gender, when a patient complains of unexplained nausea, abdominal pain, or vomiting. PMID- 16252278 TI - Help seeking and barriers to treatment in a community sample of Mexican American and European American women with eating disorders. AB - OBJECTIVE: The study examined treatment seeking for eating disorders in Mexican American and European American women. METHOD: One hundred forty-five women with eating disorders (76 Mexican American, 69 European American) were diagnosed using the Structured Clinical Interview for DSM-IV-TR (SCID-IV) and Eating Disorder Examination (EDE). RESULTS: Only 28% of the sample reported having sought treatment for their eating problems and only 17% had received treatment. Both groups were equally likely to believe they have significant eating problems and to want help. However, Mexican Americans were less likely to have sought treatment and, having sought help, were less likely to have been diagnosed or treated. European Americans were more likely to have utilized psychotherapists, psychiatrists, and psychotropic medications, whereas Mexican Americans largely had sought help from general practitioners for weight concern. The two groups endorsed similar barriers to treatment seeking. CONCLUSION: Results support clinical impressions that eating disorders largely go undetected and untreated. Nonspecialists may be likely to fail to detect eating disorders. PMID- 16252280 TI - Massive gastric dilatation after a single binge in an anorectic woman. AB - OBJECTIVE: Massive gastric dilatation is a very serious condition that is extremely rare in patients with no history of gastrointestinal disease. Several cases have been reported in patients with eating disorders, particularly after a binge. We report here the case of a young woman who developed severe gastric dilatation after a single binge. METHODS: A computed tomographic (CT) scan of the abdomen was done and a psychiatric evaluation was performed. RESULTS: The diagnosis of acute gastric dilatation was confirmed and superior mesenteric artery syndrome was excluded. The patient responded to nasogastric drainage and bowel rest. She was also found to have situational anxiety and depressive symptoms as well as a nonspecified eating disorder. CONCLUSION: This case illustrates the serious sequel of even a single binge in any patient with abnormal dietary habits, and demonstrates the useful role of the CT scan in the diagnosis. PMID- 16252281 TI - Zebrafish id2 developmental expression pattern contains evolutionary conserved and species-specific characteristics. AB - The inhibitor of differentiation or inhibitor of DNA binding (Id) family are members of the helix-loop-helix (HLH) group of transcription factors that play important roles in cell proliferation, differentiation, cell cycle control, and apoptosis. They modulate the formation of active class A-class B basic HLH (bHLH) complexes. Ids lack the amino-terminal associated basic region necessary for DNA binding, thus sequestering the class A factors, inhibiting the formation of active class A-class B heterodimers and, therefore, are considered to act as dominant-negative regulators of differentiation pathways. We isolated zebrafish id2, and its expression during development was characterized. id2, in addition to regions of expression detected in Xenopus and mice, is also expressed in the tegmentum; midbrain-hindbrain boundary; cerebellum; rhombomeres 2,3,4,6; notochord; and corpuscles of Stannius. Furthermore, we show that expression of id2 is repressed in mind bomb mutants, suggesting a role of Notch upstream of Id2. PMID- 16252283 TI - Knowledge-based voting algorithm for automated protein functional annotation. AB - Automated annotation of high-throughput genome sequences is one of the earliest steps toward a comprehensive understanding of the dynamic behavior of living organisms. However, the step is often error-prone because of its underlying algorithms, which rely mainly on a simple similarity analysis, and lack of guidance from biological rules. We present herein a knowledge-based protein annotation algorithm. Our objectives are to reduce errors and to improve annotation confidences. This algorithm consists of two major components: a knowledge system, called "RuleMiner," and a voting procedure. The knowledge system, which includes biological rules and functional profiles for each function, provides a platform for seamless integration of multiple sequence analysis tools and guidance for function annotation. The voting procedure, which relies on the knowledge system, is designed to make (possibly) unbiased judgments in functional assignments among complicated, sometimes conflicting, information. We have applied this algorithm to 10 prokaryotic bacterial genomes and observed a significant improvement in annotation confidences. We also discuss the current limitations of the algorithm and the potential for future improvement. PMID- 16252284 TI - Structural differences between allelic variants of the ovine prion protein revealed by molecular dynamics simulations. AB - We have modeled ovine prion protein (residues 119-233) based on NMR structures of PrP from other mammalian species. Modeling of the C-terminal domain of ovine PrP predicts three helices: helix-1 (residues 147-155), flanked by two short beta strands; helix-2 (residues 176-197), and helix-3 (residues 203-229). Molecular dynamics simulations on this model of ovine PrP have determined structural differences between allelic variants. At neutral pH, limited root mean-squared (RMS) fluctuations were seen in the region of helix-1; between beta-strand-2 and residue 171, and the loop connecting helix-2 and helix-3. At low pH, these RMS fluctuations increased and showed allelic variation. The extent of RMS fluctuation between beta-strand 2 and residue 171 was ARR > ARQ > VRQ. This order was reversed for the loop region connecting helix-2 and helix-3. Although all three variants have the potential to display an extended helix at the C-terminal region of helix-1, the major influence of the VRQ allele was to restrict the conformations of the Asn162 and Arg139 side-chains. Variations observed in the simulations in the vicinity of helix-1 correlated with reactivity of C-terminal specific anti-PrP monoclonal antibodies with peripheral blood cells from scrapie susceptible and -resistant genotypes of sheep: cells from VRQ homozygous sheep showed uniform reactivity, while cells from ARQ and ARR homozygous sheep showed variable binding. Our data show that molecular dynamics simulations can be used to determine structural differences between allelic variants of ovine PrP. The binding of anti-PrP monoclonal antibodies to ovine blood cells may validate these structural predictions. PMID- 16252282 TI - Tyrosine hydroxylase gene regulation in human neuronal progenitor cells does not depend on Nurr1 as in the murine and rat systems. AB - A previous study on the human tyrosine hydroxylase (TH) promoter revealed remarkable differences in the mechanism of TH gene regulation between the human and murine models. Indeed, a low degree of homology was observed in the sequence of TH promoters among human, mouse, and rat systems. Only five short conserved regions (CRs) could be identified among the three species. A human TH minimal promoter was engineered and assembled into a self-inactivating lentiviral vector system. This human TH minimal promoter contained the five CRs plus the first -194 bp from the transcription start of the human TH promoter and the first 35 bp of the untranslated messenger RNA leader of the human TH gene. A significant degree of specificity for this human TH minimal promoter was observed only for human neuronal progenitor cells (hNPCs), but not for TH-positive differentiated mouse primary striatal and substantia nigra cells, indicating a significant difference in TH gene regulation between the human and mouse systems. Not only is the degree of homology between the human and mouse promoters in the range of only 46%, but also those few elements that share a high degree of homology display totally different functions in human and mouse brain-derived cells. In the rodent system, NR4A2 (Nurr1) is required for the transactivation of TH minimal promoters. Intriguingly, neither the dimeric nor the heterodimeric binding sites for Nurr1 are present in the 13 kb DNA sequence that contains the human TH promoter. Instead, the CRs termed one and four of the human TH promoter encode only for a half palindromic binding site sequence for Nurr1, which failed to bind Nurr1 in an in vitro electrophoretic mobility shift assay (EMSA). Additionally, of the three monomeric NGFI-B response element (NBRE) core sites (AGGTCA) and two NBRE related sites present in the human TH promoter, only one core and two NBRE related sites formed protein binding complexes. Interestingly, there was no increase of protein binding complex formation upon TH induction and in no case could antibodies supershift Nurr1 from the complex. These findings, taken together, demonstrate that NBRE-related binding sites for Nurr1 do not play a direct role in mediating an interaction between Nurr1 and the human TH promoter. Likewise, immunohistochemical and Western blot analysis have also confirmed that both endogenous and exogenous Nurr1 expression does not positively correlate with TH gene expression in hNPCs, in contrast to the mouse model. In addition, real time PCR analysis revealed that the downregulation of human Nurr1 gene expression mediated by silencing RNA molecules did not affect human TH gene expression in differentiated hNPCs. A better understanding of human TH gene regulation may have important implications both for the development of novel therapeutic approaches and the study of the pathogenesis of a variety of neurological illnesses, including Parkinson's disease, bipolar disorder, and schizophrenia. PMID- 16252285 TI - Interactions between sleeping position and feeding on cardiorespiratory activity in preterm infants. AB - Infants sleeping in the prone position are at greater risk for sudden infant death syndrome (SIDS). Sleep position-dependent changes in cardiorespiratory activity may contribute to this increased risk. Cardiorespiratory activity is also affected by feeding. Twenty prematurely-born infants were studied at 31-36 weeks postconceptional age while sleeping in the prone and supine positions. Heart rate, respiratory rate, and patterns of variability were recorded during interfeed intervals, and effects of position and time after feeding were analyzed by repeated measures analyses of variance. There were significant effects of both sleeping position and time after feeding. Heart rate is higher and heart period variability is lower in the prone position, and the effects of sleeping position on cardiac functioning are more pronounced during the middle of the intrafeed interval. In preterm infants, autonomic responses to nutrient processing modulate the cardiorespiratory effects of sleeping position. Prone sleeping risk may vary with time after feeding. PMID- 16252286 TI - Fishing for the secrets of vertebrate evolution in threespine sticklebacks. AB - The threespine stickleback (Gasterosteus aculeatus) is rapidly emerging as a new model genetic system to study questions at the interface of evolution and development. The relatively rapid and recent diversification of this small teleost fish, combined with the development of genetic and genomic tools for this fish, provides an unprecedented opportunity to identify the genetic and molecular basis of morphological variation in natural populations of vertebrates. Recently, the genes underlying two different adaptive morphological traits in stickleback have been identified. This work has provided answers to four longstanding questions in the field of evolution and development: (1) How many genes underlie morphological variation in natural populations? (2) What are the genes that underlie morphological variation in natural populations? (3) Do coding or regulatory mutations underlie morphological evolution? (4) What is the molecular and genetic basis of parallel morphological evolution? Because stickleback populations also display natural variation in morphology, life history, physiology, and behavior, extending the approaches used to identify the genetic basis of morphological variation in sticklebacks to other phenotypes is sure to yield further important insights into the genetic and developmental basis of diversity in natural populations. PMID- 16252288 TI - Myron Hofer: an appreciation. PMID- 16252289 TI - Developmental psychobiology for public health: a bridge for translational research. PMID- 16252290 TI - Mother and newborn baby: mutual regulation of physiology and behavior--a selective review. AB - This article reviews 30 years of work demonstrating that interactions between mother and newborn infant in the period just after birth influence the physiology and behavior of both. Close body contact of the infant with his/her mother helps regulate the newborn's temperature, energy conservation, acid-base balance, adjustment of respiration, crying, and nursing behaviors. Similarly, the baby may regulate--i.e., increase--the mother's attention to his/her needs, the initiation and maintenance of breastfeeding, and the efficiency of her energy economy through vagus activation and a surge of gastrointestinal tract hormone release resulting in better exploitation of ingested calories. The effects of some of these changes can be detected months later. Parallels to animal research and possible mechanisms are discussed. PMID- 16252292 TI - Development and evolution of hidden regulators: selective breeding for an infantile phenotype. AB - Mother-infant separation in the rat has been used as an analytical tool to reveal biosocial processes underlying infant physiology and behavior. The same strategy has guided a project in which selective breeding for an infantile behavior has provided insights into how biological systems become recruited and integrated as expressions of temperamental affective responses. Two lines of rats (High and Low USV lines) were selectively bred based on rates of USV emission to maternal separation and isolation at postnatal day (P) 10. After many generations of breeding, the High and Low lines show widespread and distinctly different profiles of physiology and behavior in the first 3 weeks of life. Insights gained from longitudinal studies suggest that selection may work by reorganizing developmental processes, not just a given trait, over the postnatal period. As animal models, the lines have the potential to provide valuable tools for understanding developmental mechanisms underlying genetic and developmental risk for depression/anxiety syndromes in children and adults. PMID- 16252293 TI - Attachment, loss, and complicated grief. AB - Bereavement is a highly disruptive experience that is usually followed by a painful but time-limited period of acute grief. An unfortunate minority of individuals experience prolonged and impairing complicated grief, an identifiable syndrome that differs from usual grief, major depression, and other DSM IV diagnostic entities. Underlying processes guiding symptoms are not well understood for either usual or complicated grief. We propose a provisional model of bereavement, guided by Myron Hofer's question "What exactly is lost when a loved one dies?" We integrate insights about biobehavioral regulation from Hofer's animal studies of infant separation, research on adult human attachment, and new ideas from bereavement research. In this model, death of an attachment figure produces a state of traumatic loss and symptoms of acute grief. These symptoms usually resolve following revision of the internalized representation of the deceased to incorporate the reality of the death. Failure to accomplish this integration results in the syndrome of complicated grief. PMID- 16252294 TI - Blood pressure responses to feeding in infancy: spin-offs of serendipity. AB - This article summarizes results from a program of research that has focused on understanding the characteristics of, and factors that shape, acute cardiovascular responses to feeding in infants. The work developed from behavioral observations in rats suggesting a linkage between variations in maternal behavior and adult blood pressure. However, the hypothesis that specific types of interactions, in particular those associated with feeding, might alter cardiovascular function was greatly influenced by a serendipitous observation made by Myron Hofer several years earlier. Myron discovered that nutrient availability is a key regulator of cardiovascular function in newborn rats. Our spin-off studies established that, in newborn rats and human infants, ingestion of nutrient is associated with robust increases in heart rate and blood pressure. PMID- 16252295 TI - Guidance and differentiation of maternally directed orienting behaviors: probing the origins of filial attachment. AB - Newborn rats seek and attain contact with their mothers by orienting and primitive locomotor behaviors called maternally-directed orienting behaviors (MDOBs). By 2 days of age MDOBs are elicited specifically by maternal features and are heightened by a period of acute separation from the mother. This behavioral regulation by the mother's presence suggests an early form of filial attachment. To probe for other evidence of filial attachment, two aspects of the controls of MDOBs were investigated: (1) whether MDOBs are goal-corrected in relation to the dam's position and (2) whether the performance of MDOBs can be separated from that of suckling. It was found that, during the first 2 days after birth, MDOBs maintained the pup's ventrum-to-ventrum contact with the dam regardless of her position, suggesting that they are goal-corrected. Evidence was not found for independent controls of nipple grasping (NG), the behavior that initiates suckling, and MDOBs during the first two postnatal days. PMID- 16252291 TI - Neurobiology of infant attachment. AB - A strong attachment to the caregiver is critical for survival in altricial species, including humans. While some behavioral aspects of attachment have been characterized, its neurobiology has only recently received attention. Using a mammalian imprinting model, we are assessing the neural circuitry that enables infant rats to attach quickly to a caregiver, thus enhancing survival in the nest. Specifically, the hyper-functioning noradrenergic locus coeruleus (LC) enables pups to learn rapid, robust preference for the caregiver. Conversely, a hypo-functional amygdala appears to prevent the infant from learning aversions to the caregiver. Adult LC and amygdala functional emergence correlates with sensitive period termination. This study suggests the neonatal brain is not an immature version of the adult brain but is uniquely designed to optimize attachment to the caregiver. Although human attachment may not rely on identical circuitry, the work reviewed here suggests a new conceptual framework in which to explore human attachments, particularly attachments to abusive caregivers. PMID- 16252296 TI - Protein flexibility and ligand rigidity: a thermodynamic and kinetic study of ITAM-based ligand binding to Syk tandem SH2. AB - The Syk tandem Src homology 2 domain (Syk tSH2) constitutes a flexible protein module involved in the regulation of Syk kinase activity. The Syk tSH2 domain is assumed to function by adapting the distance between its two SH2 domains upon bivalent binding to diphosphotyrosine ligands. A thermodynamic and kinetic analysis of ligand binding was performed by using surface plasmon resonance (SPR). Furthermore, the effect of binding on the Syk tSH2 structural dynamics was probed by hydrogen/deuterium exchange and electrospray mass spectrometry (ESI MS). Two ligands were studied: 1, a flexible peptide derived from the tSH2 recognition ITAM sequence at the gamma chain of the FcepsilonRI-receptor, and 2, a ligand in which the amino acids between the two SH2 binding motifs in ligand 1 have been replaced by a rigid linker of comparable length. Both ligands display comparable affinity for Syk tSH2 at 25 degrees C, yet a major difference in thermodynamics is observed. Upon binding of the rigid ligand, 2, the expected entropy advantage is not realized. On the contrary, 2 binds with a considerably higher entropy price of approximately 9 kcal mol-1, which is attributed to a further decrease in protein flexibility upon binding to this rigid ligand. The significant reduction in deuterium incorporation in the Syk tSH2 protein upon binding of either 1 or 2, as monitored by ESI-MS, indicates a major reduction in protein dynamics upon binding. The results are consistent with a two-step binding model: after an initial binding step, a rapid structural change of the protein occurs, followed by a second binding step. Such a bivalent binding model allows high affinity and fast dissociation kinetics, which are very important in transient signal-transduction processes. PMID- 16252298 TI - Carbohydrate array analysis of anti-Tn antibodies and lectins reveals unexpected specificities: implications for diagnostic and vaccine development. AB - The Tn antigen is a carbohydrate antigen expressed in most carcinomas, during embryogenesis, on pathogenic parasites, and on HIV. It has been evaluated extensively as a potential diagnostic marker and several Tn-based vaccines are in clinical trials. Based on discrepancies in the literature regarding Tn expression, we began to question whether antibodies and lectins used routinely to detect the Tn antigen were providing accurate information. To investigate this possibility, a carbohydrate microarray and a highly sensitive assay were developed and three frequently used Tn receptors (HBTn1, Bric111, and VVL-B4) were evaluated. Carbohydrate-array analysis revealed unexpected cross-reactivity with other human carbohydrate epitopes. VVL-B4 bound the Tn antigen, GalNAcalpha1 6Gal, and GalNAcalpha1-3Gal. Bric111 bound the Tn antigen, blood group A, GalNAcalpha1-6Gal, and GalNAcalpha1-3Gal. HBTn1 showed the best selectivity, but still displayed moderate binding to blood group A. Implications for the development of Tn-based diagnostics and vaccines are discussed. PMID- 16252299 TI - Structural transition and tuning of tert-butylamine hydrate. PMID- 16252297 TI - Pityriarubins, novel highly selective inhibitors of respiratory burst from cultures of the yeast Malassezia furfur: comparison with the bisindolylmaleimide arcyriarubin A. AB - Pityriasis versicolor is the most common skin mycosis in humans worldwide. Yeasts of the genus Malassezia, particularly M. furfur, a saprophyte occurring widely on human skin, are generally regarded as the causative agents. M. furfur is able to convert tryptophan into a variety of indole alkaloids, some of them showing biological properties that correlate well with certain clinical features of pityriasis versicolor. This suggests a possible role for these compounds in the pathophysiology of the disease. We here report that the novel pityriarubins A, B and C, isolated from cultures of the yeast, inhibit respiratory burst in human neutrophils, activated by various agents, in a highly selective, unexpected manner. The release of 5-lipoxygenase products after challenge of neutrophils with the calcium ionophore A23187 is also inhibited in a dose-dependent manner. These activities reflect the close structural relationship of pityriarubins to bisindolylmaleimides, which have recently gained great interest as protein kinase inhibitors. PMID- 16252300 TI - Stephacidin B, the avrainvillamide dimer: a formidable synthetic challenge. PMID- 16252301 TI - Molecular AND logic gate based on electric dichroism of a photochromic dihydroindolizine. PMID- 16252302 TI - Ethylene oligomerization at coordinatively and electronically unsaturated low valent nickel. PMID- 16252303 TI - Kinetic resolution of chiral secondary alcohols by dehydrogenative coupling with recyclable silicon-stereogenic silanes. PMID- 16252304 TI - Efficient enrichment of intact phosphorylated proteins by modified immobilized metal-affinity chromatography. AB - Phosphoproteome studies are hampered by the lack of methods which allow a comprehensive and fast analysis of intact phosphoproteins. Here we describe an immobilized metal-affinity chromatography (IMAC)-based technique for the enrichment of phosphorylated proteins, which allows recovery of up to 90% of phosphoproteins. This technique is compatible with 2-DE and can be applied to cultured cells and tissues. PMID- 16252305 TI - Differential detection of S100A8 in transitional cell carcinoma of the bladder by pair wise tissue proteomic and immunohistochemical analysis. AB - The search for novel molecular markers of tumor invasion is vital if strategies are to become more effective in the diagnostic and prognostic management of transitional cell carcinoma of the bladder. Up to 50% of tumors detected at stage 1 (pT1) progress to a higher grade even after endoscopic surgical resection, and there are currently no protein markers of this aggressive, invasive phenotype. We have combined SELDI-TOF-MS, ClinProt magnetic bead enrichment, Nano-LC-ESI-ion trap tandem mass spectrometry and immunohistochemical analysis to the study of 12 invasive bladder cancer tissue biopsies paired with normal bladder tissue samples obtained from the same patients for the definition and identification of proteins up-regulated in the tumors. We report the inflammation-associated calcium binding protein S100A8 (MRP-8, calgranulin A) to be highly expressed in tumor cells in contrast to normal urothelium in 50% of the samples, as well as two unidentified protein markers at 5.75 and 6.89 kDa that were differentially detected in 9/12 and 10/12 tumor samples, respectively. These new markers, when fully characterized, may contribute to new target proteins for the prediction of aggressive, invasive bladder tumors. PMID- 16252307 TI - Proteome analysis of cortical neuronal cultures following cycloheximide, heat stress and MK801 preconditioning. AB - Studying endogenous neuroprotective mechanisms induced by preconditioning may provide drug leads to reduce ischemic neuronal death. In this study, we used 2-DE to examine protein expression following cycloheximide, heat stress, and MK801 preconditioning in rat cortical neuronal cultures. Of 150 differentially expressed protein spots selected for identification the protein or tentative protein(s) were identified in 84 cases, representing 50 different proteins. Different protein spots representing the same protein or closely related protein(s) occurred for 21 of the identified proteins and are likely to represent PTMs or proteolytic fragments of the protein. Six protein spots (actin, elongation factor 1-alpha 1, peptidyl-prolyl cis-transisomerase A, Cu/Zn superoxide dismutase, stathmin, tropomyosin) were differentially expressed in all three preconditioning treatments. Twenty-seven protein spots were differentially expressed in two preconditioning treatments, while 51 spots were differentially expressed in one treatment. Three proteins heterogeneous nuclear ribonucleoproteins A2/B1, mitochondrial stress-70 protein, and tropomyosin were detected in control neuronal cultures, but not following one or more preconditioning treatments, while a posttranslational modified form of the voltage dependent anion channel 1 was only detected following cycloheximide preconditioning. In summary, this study has revealed multiple protein changes potentially involved in neuroprotective and neurodamaging pathways, which require further characterization. PMID- 16252306 TI - Oxidation of specific methionine and tryptophan residues of apolipoprotein A-I in hepatocarcinogenesis. AB - Hepatocellular carcinoma (HCC) is the fifth most common neoplasm with more than 500 000 new cases diagnosed yearly. Although major risk factors of HCC are currently known, the identification of biological targets leading to an early diagnosis of the disease is considered one of the priorities of clinical hepatology. In this work we have used a proteomic approach to identify markers of hepatocarcinogenesis in the serum of a knockout mice deficient in hepatic AdoMet synthesis (MAT1A(-/-)), as well as in patients with HCC. Three isoforms of apolipoprotein A-I (Apo A-I) with different pI were identified in murine serum. Isoform 1 is up-regulated in the serum of MAT1A(-/-) mice much earlier than any histological manifestation of liver disease. Further characterization of the differential isoform by electrospray MS/MS revealed specific oxidation of methionine 85 and 216 to methionine sulfoxide while the sequence of the analogous peptides on isoforms 2 and 3 showed the nonoxidized methionine residues. Enrichment of an acidic isoform of Apo A-I was also assessed in the serum of hepatitis B virus patients who developed HCC. Specific oxidation of methionine 112 to methionine sulfoxide and tryptophans 50 and 108 to formylkinurenine were identified selectively in the up-regulated isoform. Although it is not clear at present whether the occurrence of these modifications has a causal role or simply reflects secondary epiphenomena, this selectively oxidized Apo A-I isoform may be considered as a pathological hallmark that may help to the understanding of the molecular pathogenesis of HCC. PMID- 16252308 TI - Proteome imaging: a closer look at life's organization. AB - Imaging the proteome is a term that is used in many different contexts. The term implies that the entire cohort of proteins and their modifications are visualized. This unfortunately is not the case. In this mini-review, a concise overview is provided on different imaging technologies that are currently used to investigate the structure, function and dynamics of proteins and their organization. These techniques have been selected for review based on the unique insights they provide in subsets of the proteome. These techniques have been illustrated with practical examples of their merits. Mass spectrometry-based imaging technologies are playing a key role in proteome research and have been reviewed in more detail. They hold the promise of detailed molecular insight in the spatial organization of living system. PMID- 16252309 TI - Colonic availability of apple polyphenols--a study in ileostomy subjects. AB - Nutrition is thought to play an essential role in the pathogenesis of inflammatory and malignant gastrointestinal diseases. It is well known that plant ingredients such as polyphenols and flavonoids show anticarcinogenic effects both in vitro and in animal experiments, and may thus reduce the risk of colorectal cancer in man. The aim of the study was to determine the amount of polyphenols reaching the colon after oral intake of apple juice. After consumption of a polyphenol-free diet 11 healthy ileostomy volunteers drank 1 L of a polyphenol rich cloudy apple juice. Ileostomy effluent was collected immediately before and 1, 2, 4, 6, and 8 h after consumption of apple juice. A broad spectrum of polyphenols was identified using HPLC-diode array detection (HPLC-DAD) as well as HPLC-ESI-MS/MS; quantitation was performed with HPLC-DAD. Most of the orally administered apple polyphenols were absorbed from or metabolized in the small intestine. Between 0 and 33% of the oral dose was recovered in the ileostomy bags with a maximum of excretion after 2 h. Phloretin glucuronide as product of polyphenol metabolism was detected in the ileostomy effluent. The present results show that most of the apple juice polyphenols are absorbed in the small intestine. Minor amounts of unmetabolized polyphenols are recovered in the ileostomy effluent, which would reach the colon under physiologic circumstances. These data have to be considered when polyphenols are used in model systems to show preventive effects in colorectal carcinogenesis. PMID- 16252310 TI - Solid-phase permethylation of glycans for mass spectrometric analysis. AB - A miniaturized approach was developed for quantitative permethylation of oligosaccharides, which involves packing of sodium hydroxide powder in microspin columns or fused-silica capillaries (500 microm i.d.), permitting effective derivatization in less than a minute at microscale. Prior to mass spectrometry, analytes are mixed with methyl iodide in dimethyl sulfoxide solution containing traces of water before infusing through the microreactors. This procedure minimizes oxidative degradation and peeling reactions and avoids the need of excessive clean-up. Picomole amounts of linear and branched, sialylated and neutral glycan samples were rapidly and efficiently permethylated by this approach and analyzed by matrix-assisted laser desorption/ionization mass spectrometry. PMID- 16252311 TI - Association between quality of life scores and short-term outcome after surgery for cancer of the oesophagus or gastric cardia. AB - BACKGROUND: Evidence suggests that baseline quality of life (QOL) scores are independently prognostic for survival in patients with cancer, but the role of QOL data in predicting short-term outcome after surgery is uncertain. This study assessed the association between QOL scores and short-term outcomes after surgery for oesophageal and gastric cancer. METHODS: Consecutive patients selected for oesophagectomy or total gastrectomy between November 2000 and May 2003 completed the European Organization for Research and Treatment of Cancer's quality of life questionnaire, QLQ-C30. Multivariable regression models, adjusting for known clinical risk factors, were used to investigate relationships between QOL scores, major morbidity, hospital stay and survival status at 6 months. RESULTS: Of 130 patients, 121 completed the questionnaire (response rate 93.1 per cent). There were 29 major complications (24.0 per cent) and 22 patients (18.2 per cent) died within 6 months of operation. QOL scores were not associated with major morbidity but were significantly related to survival status at 6 months after adjusting for known clinical risk factors. A worse fatigue score of 10 points (scale 0-100) corresponded to an increase in the odds of death within 6 months of surgery of 37.4 (95 per cent confidence interval (c.i.) 12.4 to 67.8) per cent (P = 0.002). Pretreatment social function scores were moderately associated with hospital stay (P = 0.021); a reduction in social function by 10 points corresponded to an increase in hospital stay of 0.93 (95 per cent c.i. 0.12 to 1.74) days. CONCLUSION: QOL scores supplement standard staging procedures for oesophageal and gastric cancer by providing prognostic information, but they do not contribute to perioperative risk assessment. PMID- 16252312 TI - Long-term results in patients with T2-3 N0 distal rectal cancer undergoing radiotherapy before transanal endoscopic microsurgery. AB - BACKGROUND: Local excision after radiotherapy for node-negative low rectal cancer may be an alternative to radical excision. This study evaluated the results of local excision in patients with small (less than 3 cm in diameter) T2 and T3 distal rectal tumours following neoadjuvant therapy. METHODS: One hundred patients with rectal cancer (54 uT2 and 46 uT3 uN0 tumours) were enrolled. All patients underwent preoperative radiotherapy followed by local excision by means of transanal endoscopic microsurgery. RESULTS: Definitive histological examination revealed nine pT1, 54 pT2 and 19 pT3 tumours. A complete response (R0) or microscopic residual tumour (R1mic) was found in three and 15 patients respectively. Minor complications occurred in 11 patients and major complications in two. At a median follow-up of 55 (range 7-120) months, the local failure rate was 5 per cent and metastatic disease was found in two patients. The cancer specific survival rate at 90 months' follow-up was 89 per cent, and the overall survival rate 72 per cent. Salvage abdominoperineal resection was performed in three patients, two of whom were disease free at 15 and 19 months. CONCLUSION: Treatment of small uT2 and uT3 uN0 rectal cancers with preoperative high-dose radiotherapy followed by transanal endoscopic microsurgery is an acceptable alternative to conventional radical resection. PMID- 16252313 TI - Systematic review of randomized trials comparing rubber band ligation with excisional haemorrhoidectomy. AB - BACKGROUND AND METHOD: This review compares the two most popular treatments for haemorrhoids, namely rubber band ligation (RBL) and excisional haemorrhoidectomy. Randomized trials were identified from the major electronic databases. Symptom control, retreatment, postoperative pain, complications, time off work and patient satisfaction were assessed. Relative risk (RR) and weighted mean difference with 95 per cent confidence interval (c.i.) were estimated using a random-effects model for dichotomous and continuous outcomes respectively. RESULTS: Three trials met the inclusion criteria and all were of poor methodological quality. Complete remission of haemorrhoidal symptoms was better after haemorrhoidectomy (RR 1.68 (95 per cent c.i 1.00 to 2.83)). There was significant heterogeneity between the studies (I(2) = 90.5 per cent; P < 0.001). Fewer patients required retreatment after haemorrhoidectomy (RR 0.20 (95 per cent c.i 0.09 to 0.40)), but anal stenosis, postoperative haemorrhage and incontinence to flatus were more common with this operation. CONCLUSIONS: Haemorrhoidectomy produced better long-term symptom control in patients with grade III haemorrhoids, but was associated with more postoperative complications than RBL. PMID- 16252314 TI - Systematic review and meta-analysis of the effectiveness of antibiotic prophylaxis in prevention of wound infection after mesh repair of abdominal wall hernia. AB - BACKGROUND: The aim was to determine whether systemic antibiotic prophylaxis prevented wound infection after repair of abdominal wall hernia with mesh. METHODS: This was a systematic review of the available literature identified from multiple databases using the terms 'hernia' and 'antibiotic prophylaxis'. Randomized placebo-controlled trials of antibiotic prophylaxis in abdominal wall mesh hernia repair with explicitly defined wound infection criteria and a minimum follow-up of 1 month were included. After independent quality assessment and data extraction, data were pooled for meta-analysis using a random-effects model. RESULTS: The search process identified eight relevant trials. Two papers on umbilical, incisional or laparoscopic hernias, and six concerning inguinal and femoral (groin) hernias were suitable for meta-analysis. The incidence of infection after groin hernia repair was 38 (3.0 per cent) of 1277 in the placebo group and 18 (1.5 per cent) of 1230 in the antibiotic group. Antibiotic prophylaxis did not significantly reduce the incidence of infection: odds ratio 0.54 (95 per cent confidence interval 0.24 to 1.21); number needed to treat was 74. The number of deep infections was six (0.6 per cent) in the placebo group and three (0.3 per cent) in the antibiotic prophylaxis group: odds ratio 0.50 (95 per cent c.i. 0.12 to 2.09). CONCLUSION: Antibiotic prophylaxis did not prevent the occurrence of wound infection after groin hernia surgery. More trials are needed for complete evidence in other areas of abdominal wall hernia. PMID- 16252315 TI - Rules governing protein identification by mass spectrometry. PMID- 16252317 TI - Multivariate optimization approach for chiral resolution of drugs using human serum albumin in affinity electrokinetic chromatography-partial filling technique. AB - The enantiomeric resolution of chiral compounds using HSA by means of affinity EKC (AEKC)-partial filling technique is the result of a delicate balance between different experimental variables such as protein concentration, running pH (background electrophoretic buffer, protein and compound solutions) and protein solution plug length. In this paper multivariate optimization approaches for chiral separation of four basic drugs (alprenolol, oxprenolol, promethazine and propranolol) using HSA as chiral selector in AEKC-partial filling technique are studied. The experimental conditions to achieve maximum resolution are optimized using the Box-Behnken experimental design. Partial least squares and pareto charts are used to analyse the main effects on the resolution. The experimental resolutions observed for all compounds studied in optimum conditions agree with the estimated values based on response surface models. The results obtained show that the range of experimental conditions that provided enantioresolution narrows as hydrophobicity of analytes decreases. This fact can be explained by assuming that hydrophobicity controls the interaction of basic compounds with HSA. PMID- 16252318 TI - Glycine-based polymeric surfactants with varied polar head group: II. chemical selectivity in micellar electrokinetic chromatography using linear solvation energy relationships. AB - A series of four acyl and four alkenoxy glycinates (i.e., mono-, di-, tri-, and tetraderivatives of polysodium N-undecenoyl glycinate (poly-SUGs) as well as polysodium N-undecenoxy carbonyl glycinates (poly-SUCGs)) were compared for simultaneous separation of nonhydrogen bonding (NHB), hydrogen-bond acceptor (HBA), and hydrogen-bond donor (HBD) solutes. An increase in the number of glycine units in the polar head group of polymeric surfactant decreases both the retention and the migration window of all solutes with some changes in separation selectivity. The poly(sodium N-undecenoxy carbonyl-glycinate) (poly-SUCG1) with one glycine unit was the least polar surfactant and has the lowest phase ratio, but this monoglycinate surfactant provided the best simultaneous separation of 10 NHBs and 8-HBAs. On the other hand, 9-HBDs were well separated using any of the six mono-, di-, and triglycinate surfactants compared to the two tetraglycinates. Linear solvation energy relationships (LSERs) and separation of the geometrical isomers studies were also performed to further envisage the selectivity differences. From LSER studies, the phase ratio and hydrogen-bond-donating strength of the poly-SUG series of surfactant were found to increase with an increase in the size of the head group, but no clear trends were observed for poly-SUCG surfactants. The cohesiveness for all poly-SUG and poly-SUCG was positive, but the values were generally lower (with exception of the poly(sodium N-undecenoyl glycyl-glycyl-glycinate)) at a higher number of glycine units. Finally, the poly(sodium N-undecenoyl glycinate) and poly-SUCG1 were found to be the two best polymeric surfactants as they provided relatively higher shape selectivity for separation of two of the three sets of geometrical isomers. PMID- 16252322 TI - Quantitation in capillary electrophoresis-mass spectrometry. AB - CE-MS has evolved into a strong alternative to LC-MS. Most of CE-MS applications deal with characterization and identification. However, quantitative aspects have gained importance in, e.g., pharmaceutical and biotechnological applications. Here we summarize and evaluate various methodological aspects in order to achieve sensitive and reproducible results. Similar to LC-MS, aspects of matrix influence on the electrospray process need to be carefully addressed when quantitative results are intended by CE-MS. Due to a more complicated coupling special emphasis needs to be put on the CE-MS interface. Generally linearity over more than three orders of magnitude can be achieved by CE-ESI-MS. Furthermore, a literature survey has been performed in order to give an overview over quantitative measurements performed by CE-MS. The precision can be doubled when changing from a structural related to an isotopically labeled internal standard. Thus a level of precision better than 5% RSD can be achieved. PMID- 16252319 TI - Comparison of dodecoxycarbonylvaline microemulsion, solvent-modified micellar and micellar pseudostationary phases for the chiral analysis of pharmaceutical compounds. AB - A direct comparison of dodecoxycarbonylvaline (DDCV) microemulsion, micellar and butanol-modified micellar aggregate systems was performed employing both 2 and 4% DDCV. With respect to either DDCV concentration, use of the micellar system provided the largest elution range, followed by the butanol-modified micellar system and ultimately the microemulsion. Using 2% DDCV, all three aggregate analyses yielded similar values for enantioselectivity, resolution and retention factors that were slightly better using the micellar phase, but efficiencies were consistently better using either the microemulsion or butanol-modified micellar phases. Largely, the microemulsion and butanol-modified micellar phases behaved fairly similar, although use of the butanol-modified micelle provided resolution and efficiency that were slightly better for all but two of the compounds. While reasonable separations were achieved using 2% DDCV, the results using 4% DDCV for the microemulsion system were far superior. Analyses using analogous micellar and butanol-modified micellar aggregates were unstable, making them unsuitable for use at that surfactant concentration. PMID- 16252323 TI - Metal ion speciation and capillary electrophoresis: application in the new millennium. AB - Metal ions are essential for human beings at low concentrations but they are toxic or even carcinogenic at high concentrations. Many metallic ions are found in the environment in different species which are differentiated not only by their physicochemical forms but also by their diverse toxicities with respect to living organisms (speciation). CE has been used for metal ion speciation. The present review article describes the recent trends in metal ion speciation by CE. This article deals with the speciation of metal ions, optimization of the speciation (by independent and dependent variables), hyphenation of CE, validation of the methods, mechanisms of speciation, CE versus chromatography and conclusions. PMID- 16252324 TI - Branched polymeric labels used as drag-tags in free-solution electrophoresis of ssDNA. AB - In the framework of the classical blob theory of end-labeled free-solution electrophoresis of ssDNA, and based on recent experimental data with linear and branched polymeric labels (or drag-tags), the present study puts forward design principles for the optimal type of branching that would give, for a given total number of monomers, the highest effective frictional drag for ssDNA sequencing purposes. The hydrodynamic radii of the linear and branched labels are calculated using standard models like the freely jointed chain model and the Kratky-Porod worm-like chain model. Based on comparisons of the theory with the experimental data, we propose that the design of new branched labels should use either side chains whose length is comparable to the distance between the branching points or two long branches located near the ends of the molecule's backbone. PMID- 16252325 TI - Effects of nonequilibrium on velocity and plate height in reactive capillary electrophoresis. AB - Models for velocity and plate height for reactive CE are developed under the formalism of generalized nonequilibrium theory, as described by Giddings. The resultant equations are consistent with chromatographic theory and validated with an independent stochastic simulation. Moreover, unlike prior methods for CE, this model allows calculation of thermodynamic equilibrium constants and kinetic rate constants from a single, undistorted peak. The theoretical development shows that velocity is directly dependent on the equilibrium constant and is independent of the rate constant. On the other hand, plate height varies little with equilibrium constant and is inversely proportional to rate constant. The ability to evaluate equilibrium constants from velocity and rate constants from plate height is most greatly influenced by electric field strength and mobility difference. The accuracy in calculated equilibrium constants is limited by mobility difference; however, the accuracy in rate constants is limited by plate height and equilibrium constant. PMID- 16252326 TI - Modeling of anti-Langmuirian peaks in micellar electrokinetic chromatography: benzene and naphthalene. AB - Peaks of benzene (bz) and naphthalene (np) having diffuse fronts and steep rears under overload conditions were studied quantitatively in MEKC with SDS surfactant. The retardation factors of these compounds, solubilized at microM to mM concentrations by either 10, 30, or 50 mM SDS, were determined by vacancy MEKC and frontal analysis MEKC. Isotherm coordinates were calculated from the retardation factors, and the equation for the concave upward anti-Langmuir isotherm was fit to them. Peak profiles were computed with the MacCormack algorithm from the isotherm fits and a simplified continuity equation appropriate to MEKC. These profiles were compared to ones generated in normal MEKC from samples of bz and np solubilized at muM to mM concentrations by either 10, 30, or 50 mM SDS. In all cases, the anti-Langmuir isotherm described the asymmetry of experimental peaks. For bz in 30 and 50 mM SDS and np in 10 and 50 mM SDS, good to excellent agreement was found between the experimental and predicted profiles. For bz in 10 mM SDS, the experimental profiles were more broadened than the predicted ones, although their asymmetries agreed. For np in 30 mM SDS, the experimental isotherm predicted greater peak asymmetry than was observed, and the correct anti-Langmuir isotherm for all sample concentrations and field strengths was calculated from the most asymmetrical peak by the inverse method. The relative decrease of zone velocity with increasing analyte concentration was calculated from the isotherm parameters, electrokinetic mobilities, retardation factors, surfactant concentrations, and CMC. The simplification of the continuity equation was justified. PMID- 16252327 TI - Utilizing the pH hysteresis effect for versatile and simple electrophoretic analysis of proteins in bare fused-silica capillaries. AB - The analysis of peptides and proteins by CE is often desirable due to low sample consumption and possibilities for nondenaturing yet highly effective separations. However, adsorption to the inner surfaces of fused-silica capillaries often is detrimental to such analyses. This phenomenon is especially pronounced in the analysis of basic proteins and proteins containing exposed positively charged patches. To avoid wall interactions numerous buffer additives and static and dynamic wall coating principles have been devised. We previously showed (J. Chromatogr. A 2004, 1059, 215-222) that CE of the basic protein beta2 glycoprotein was rendered possible by an acidic pretreatment step, and we attributed this observation to the so-called pH hysteresis effect that influences the time for pH equilibration of the capillary wall and thus the effective wall charge and the electroosmotic mobility. We here investigate the effects of different pretreatment techniques on EOF values and on the rate of the deprotonation of silanol groups when performing the electrophoresis at neutral pH. We show the utility of this simple approach for the CE analysis of a number of basic proteins in plain silica capillaries at physiological pH. PMID- 16252328 TI - Multicapillary electrophoresis of unlabeled DNA fragments with high-sensitive laser-induced fluorescence detection by counter-current migration of intercalation dye. AB - Analysis of PCR fragments for applications, such as screening of nucleotide polymorphisms, detection of somatic mutations, or quantification of reverse transcription PCR products, becomes central in clinical research as well as preventive testing, diagnostic screening, and pharmacogenomic genotyping. A variety of CE techniques, utilizing great potential of multicapillary-array sequencers, is now commonly applied in prevention, diagnosis, and treatment of a wide range of genetic diseases (cancer, cardiovascular, and neurodegenerative diseases, etc.). Costs of fluorescently labeled primers is often a major factor in large-scale projects requiring mutation analysis in hundreds or thousands of samples. In the present paper we introduce a simple approach of detecting unlabeled DNA fragments through intercalation without a need for adding intercalator to the separation polymer matrix. The dye is only added to the anode reservoir, and mixing with the separated DNA fragments takes place upon its migration opposite to the direction of the CE separation. Using two common intercalating dyes (ethidium bromide and SYBR Green II) we present this method as a tool for routine PCR detection and quantification. PMID- 16252329 TI - A sulfonated capillary that gives reproducible migration times for capillary zone electrophoresis and micellar electrokinetic chromatography. AB - To obtain reproducible migration times and rapid analyses of analytes, sulfonate groups were chemically introduced to the inner wall of untreated fused-silica capillary with 2-(4-chlorosulfonylphenyl)ethyltrichlorosilane. The sulfonated capillary showed relatively constant electroosmotic mobility which was greater than that obtained by an untreated fused-silica capillary over the pH range studied (pH 2-9). In both CZE and MEKC, the RSDs of the migration times of analytes with the sulfonated capillary were less than 0.2% which were significantly lower than those obtained with an untreated fused-silica capillary (0.5-3.5%). When BGE were set at pH 7.0 for CZE and MEKC, the analysis times with the sulfonated capillary were about half those obtained with an untreated fused silica capillary. These results indicate that the sulfonated capillary can provide highly reproducible and rapid analyses in CE. PMID- 16252330 TI - Titanium dioxide nanoparticles-coated column for capillary electrochromatographic separation of oligopeptides. AB - A novel column made through the condensation reaction of TiO2 nanoparticles (TiO2 NPs) with silanol groups of the fused-silica capillary is described. EOF measurements under various buffer constitutions were used to monitor the completion of reactions. The results indicated that the EOF was dependent on the interactions between buffers and the bonded TiO2 NPs. With formate/Tris buffer, EOF reversal at pH below 5 and cathodic EOF at pH above 5 were indicated. The pI of the bonded TiO2 NPs was found at approximately ph 5. Only cathodic EOF was illustrated by substituting the mobile phase with either glutamate or phosphate buffer. It was elucidated that both glutamate and phosphate buffer yield a negative charge layer on the surface of TiO2 NPs attributable to the formation of a titanium complex. The CEC performance of the column was tested with angiotensin type oligopeptides. Some parameters that would affect the retention behavior were investigated. The interactions between the bonded phases and the analytes were explicated by epitomized acid-base functional groups of the oligopepetides and the speciation of the surface oxide in different pH ranges. The average separation efficiencies of 3.1 x 10(4) plates/m is readily achieved with a column of 70 cm (50 cm) x 50 mum ID under an applied voltage of 15 kV, phosphate buffer (pH 6.0, 40 mM), and UV detection at 214 nm. PMID- 16252331 TI - A silica-based monolithic column in capillary HPLC and CEC coupled with ESI-MS or electrospray-atmospheric-pressure laser ionization-MS. AB - We describe the successful coupling of CEC and capillary HPLC with the recently developed atmospheric-pressure laser ionization (APLI) method. APLI is suitable for selectively and sensitively ionizing nonpolar aromatic compounds at ambient pressure for subsequent mass-selective detection. The polycyclic aromatic hydrocarbons used as analytes are first separated either by CEC on a silica-based monolithic column or by capillary HPLC. The eluent, along with a sheath flow, is volatilized by microelectrospray and then selectively ionized by excimer laser (KrF*) radiation via two-photon excitation. A QTOF-MS is used as mass-selective detector. This interface combination makes soft ionization of thermally labile nonpolar aromatic analytes possible. PMID- 16252332 TI - Enantiomeric separation of furan derivatives and fused polycycles by cyclodextrin modified micellar capillary electrophoresis. AB - The enantiomeric separations of highly hydrophobic furan derivatives and polycycles were performed and optimized using CD-modified micellar CE. The most effective chiral selector for the enantiomeric separation of these analytes was hydroxypropyl-gamma-CD. The effects of CD and SDS concentration and organic modifier were examined in order to optimize the separation conditions. The ratio of CD to surfactant concentration affected the enantiomeric separation significantly, with increases in the derivatized CD concentration generally enhancing resolution. Addition of an organic solvent modifier to the run buffer served to increase the analytes' solubility and enhance the separation efficiency. A highly acidic pH was necessary to effectively suppress the EOF when operating in the reverse polarity mode. PMID- 16252333 TI - Migration behavior and enantioseparation of hydrobenzoin and structurally related compounds in capillary zone electrophoresis with a dual cyclodextrin system consisting of heptakis-(2,3-dihydroxy-6-O-sulfo)-beta-cyclodextrin and beta cyclodextrin. AB - Migration behavior and enantioseparation of racemic hydrobenzoin and structurally related compounds, including benzoin and benzoin methyl ether, in CZE with a dual CD system consisting of heptakis-(2,3-dihydroxy-6-O-sulfo)-beta-CD (SI-S-beta-CD) and beta-CD as chiral selectors in the presence and absence of borate complexation at pH 9.0 were investigated. The results indicate that enantioseparation of hydrobenzoin is mainly governed by CD complexation of hydrobenzoin-borate complexes with SI-S-beta-CD when SI-S-beta-CD concentration is relatively high. Whereas CD complexation of hydrobenzoin-borate complexes with beta-CD plays a significant role in enantioseparation when SI-S-beta-CD concentration is comparatively low. The (S,S)-enantiomer of the hydrobenzoin borate complex was found to interact more strongly than the corresponding (R,R) enantiomer with both SI-S-beta-CD and beta-CD. These two types of CD show the same chiral recognition pattern, but they exhibit opposite effects on the mobility of the enantiomers of hydrobenzoin-borate complexes. Enantiomer migration reversal of hydrobenzoin occurred in the presence of borate complexation when varying the concentration of beta-CD, while keeping SI-S-beta CD at a relatively low concentration. Binding constants of the enantiomers of benzoin-related compounds to beta-CD and those of hydrobenzoin-borate complexes to SI-beta-CD were evaluated; the mobility contributions of all complex species to the effective mobility of the enantiomers of hydrobenzoin as a function of beta-CD concentration in a borate buffer were analyzed. In addition, comparative studies on the enantioseparation of benzoin-related compounds with SI-S-beta-CD and with randomly sulfate-substituted beta-CD were made. PMID- 16252334 TI - Light-emitting diode-induced fluorescence detection of native proteins in capillary electrophoresis. AB - A continuous-wave 280 nm light-emitting diode (LED) was used as the excitation source for native fluorescence detection of proteins in CE. The operating current and temperature of the LED were optimized in order to achieve high luminescence power. It was found that a forward current of 30 mA and a temperature of approximately 5 degrees C gave the best S/N. By using a set of two ball lenses to focus light from the LED, we achieved a spot of approximately 200 mum with a power of 0.1-0.2 mW on the detection window. Fluorescence was collected with a ball lens at 90 degrees angle through a bandpass filter onto a photomultiplier tube. In CZE an LOD of 20 nM for conalbumin was reached. In capillary gel electrophoresis all eight proteins from a commercial standard kit were detected with high S/N. For a 10 microg/mL total protein mixture, S/N was better than 3 for all proteins in solution. Further improvement in LOD should be possible on utilization of an LED with higher luminescence power. PMID- 16252336 TI - Development of acrylate-based monolithic stationary phases for electrochromatographic separations. AB - Organic monolithic stationary phases were synthesized in fused-silica capillaries. They were prepared by in situ polymerization under UV irradiation of various alkyl acrylates, 1,3-butanediol diacrylate, and 2-acrylamido-2-methyl-1 propanesulfonic acid in a ternary porogenic solvent. The resulting stationary phases were tested in CEC. The influence of UV irradiation energy on the resulting separative performances of the monoliths was studied. It was thus demonstrated that the use of hexyl acrylate rather than butyl acrylate and lauryl methacrylate gives highly efficient monoliths (more than 300 000 plates per meter) with optimized EOF. It was also confirmed that the mobile phase ionic strength may affect significantly the separation efficiency. The influence of the nature of the mobile phase organic modifier (ACN or methanol) on EOF, retention, efficiency, and selectivity was studied and differences were observed. Finally, the performances of monolithic stationary phases developed and optimized for CEC separations were evaluated in nanoLC. PMID- 16252338 TI - Comparative study of Anaplasma parasites in tick carrying buffaloes and cattle. AB - A comparative study on the prevalence of Anaplasma parasite was conducted on ticks carrying buffaloes and cattle. Five hundred blood samples of both animals (250 of each) were collected during February, March and April. Thin blood smears on glass slides were made, fixed in 100% methyl alcohol and examined. Microscopic examination revealed that 205 (41%) animals had Anaplasma parasites, out of which 89, 44 and 72 animals had Anaplasma marginale, Anaplasma centrale and mixed infection respectively. Infected buffaloes and cattle were 75 and 130 respectively. The infection in female was 53 and 92 in buffaloes and cattle respectively. Twenty-two and 92 blood samples of male were found positive in buffaloes and cattle respectively. Comparative study revealed that the cattle were 26.82% more susceptible than buffaloes. The parasite prevailing percentage in female of both animals was slightly higher than that of the male. This investigation was aimed at studying the comparative prevalence of Anaplasma parasite in tick carrying buffaloes and cattle. PMID- 16252339 TI - Dependence of the E. coli promoter strength and physical parameters upon the nucleotide sequence. AB - The energy of interaction between complementary nucleotides in promoter sequences of E. coli was calculated and visualized. The graphic method for presentation of energy properties of promoter sequences was elaborated on. Data obtained indicated that energy distribution through the length of promoter sequence results in picture with minima at -35, -8 and +7 regions corresponding to areas with elevated AT (adenine-thymine) content. The most important difference from the random sequences area is related to -8. Four promoter groups and their energy properties were revealed. The promoters with minimal and maximal energy of interaction between complementary nucleotides have low strengths, the strongest promoters correspond to promoter clusters characterized by intermediate energy values. PMID- 16252340 TI - Protein and hordein fraction content in barley seeds as affected by sowing date and their relations to malting quality. AB - The effect of sowing date on grain protein, hordein fraction content and malting quality of two-rowed spring barley was investigated by using ten commercial cultivars with different grain protein content and the relationships among these traits were examined. The results showed that grain protein content and B hordein content increased as the sowing date postponed and were significantly affected by sowing date, while C and D hordein contents were less influenced by sowing date. There were significant differences in grain protein and hordein fraction content among the ten cultivars. The coefficient of variation of D hordein content was much larger than that of B and C hordein contents, suggesting its greater variation caused by different sowing dates. Beta-amylase activity and diastatic power were also significantly affected by sowing date, with malt extract being less affected. Significant differences in measured malt quality were found among the ten cultivars. Grain protein was significantly correlated with B hordein and malt extract positively and negatively, respectively. There was no significant correlation between beta-amylase activity or diastatic power and grain protein content. B hordein was negatively and significantly correlated with malt extract, but no significant correlations between C hordein, D hordein and malting quality traits. PMID- 16252337 TI - Acute phase reaction and acute phase proteins. AB - A review of the systemic acute phase reaction with major cytokines involved, and the hepatic metabolic changes, negative and positive acute phase proteins (APPs) with function and associated pathology is given. It appears that APPs represent appropriate analytes for assessment of animal health. Whereas they represent non specific markers as biological effect reactants, they can be used for assessing nutritional deficits and reactive processes, especially when positive and negative acute phase variables are combined in an index. When such acute phase index is applied to separate healthy animals from animals with some disease, much better results are obtained than with single analytes and statistically acceptable results for culling individual animals may be reached. Unfortunately at present no cheap, comprehensive and easy to use system is available for assessing various acute phase proteins in serum or blood samples at the same time. Protein microarray or fluid phase microchip technology may satisfy this need; and permit simultaneous analysis of numerous analytes in the same small volume sample and enable integration of information derived from systemic reactivity and nutrition with disease specific variables. Applying such technology may help to solve health problems in various countries not only in animal husbandry but also in human populations. PMID- 16252341 TI - Batch and fed-batch production of butyric acid by clostridium butyricum ZJUCB. AB - The production of butyric acid by Clostridium butyricum ZJUCB at various pH values was investigated. In order to study the effect of pH on cell growth, butyric acid biosynthesis and reducing sugar consumption, different cultivation pH values ranging from 6.0 to 7.5 were evaluated in 5-L bioreactor. In controlled pH batch fermentation, the optimum pH for cell growth and butyric acid production was 6.5 with a cell yield of 3.65 g/L and butyric acid yield of 12.25 g/L. Based on these results, this study then compared batch and fed-batch fermentation of butyric acid production at pH 6.5. Maximum value (16.74 g/L) of butyric acid concentration was obtained in fed-batch fermentation compared to 12.25 g/L in batch fermentation. It was concluded that cultivation under fed-batch fermentation mode could enhance butyric acid production significantly (P<0.01) by C. butyricum ZJUCB. PMID- 16252342 TI - Measurement and analysis of soil nitrogen and organic matter content using near infrared spectroscopy techniques. AB - Near infrared reflectance (NIR) spectroscopy is as a rapid, convenient and simple nondestructive technique useful for quantifying several soil properties. This method was used to estimate nitrogen (N) and organic matter (OM) content in a soil of Zhejiang Province, Hangzhou County. A total of 125 soil samples were taken from the field. Ninety-five samples spectra were used during the calibration and cross validation stage. Thirty samples spectra were used to predict N and OM concentration. NIR spectra of these samples were correlated using partial least square regression. The regression coefficients between measured and predicted values of N and OM was 0.92 and 0.93, and SEP (standard error of prediction) were 3.28 and 0.06, respectively, which showed that NIR method had potential to accurately predict these constituents in this soil. The results showed that NIR spectroscopy could be a good tool for precision farming application. PMID- 16252343 TI - Effective extraction of elastase from Bacillus sp. fermentation broth using aqueous two-phase system. AB - This paper presents the evaluation of an aqueous two-phase system (ATPS) for extracting elastase produced by Bacillus sp. EL31410. The elastase and cell partition behavior in polyethylene glycol (PEG)/salt systems was investigated. The suitable system for elastase extraction was PEG/KH(2)PO(4)-K(2)HPO(4), in which elastase is mainly partitioned into the PEG-rich phase, while the cells remained in the other phase. The influence of defined system parameters (e.g. PEG molecular mass, pH, NaCl addition) on the partitioning behavior of elastase is described. The concentration of phase forming components, PEG and KH(2)PO(4) K(2)HPO(4), was optimized for elastase recovery by means of response surface methodology, and it was found that they greatly influenced extraction recovery. The optimal ATPS was 23.1% (w/w) PEG 2 000 and 11.7% (w/w) KH(2)PO(4)-K(2)HPO(4). The predicted recovery was about 89.5%, so this process is suggested to be a rapid and convenient method for elastase extraction. PMID- 16252344 TI - Identification of rice seed varieties using neural network. AB - A digital image analysis algorithm based color and morphological features was developed to identify the six varieties (ey7954, syz3, xs11, xy5968, xy9308, z903) rice seeds which are widely planted in Zhejiang Province. Seven color and fourteen morphological features were used for discriminant analysis. Two hundred and forty kernels used as the training data set and sixty kernels as the test data set in the neural network used to identify rice seed varieties. When the model was tested on the test data set, the identification accuracies were 90.00%, 88.00%, 95.00%, 82.00%, 74.00%, 80.00% for ey7954, syz3, xs11, xy5968, xy9308, z903 respectively. PMID- 16252345 TI - Use of fluorometry for determination of skim milk powder adulteration in fresh milk. AB - A FAST (fluorescence of advanced Maillard products and Soluble Tryptophan) method for identification of reconstituted milk made from skim milk powder in the fresh milk was developed. Considering milk and skim milk powders variations from different seasons and countries, milk was collected from different dairy farms in different seasons and skim milk powders were collected from different countries to measure the Tryptophan (Trp), advanced Maillard products (AMP) fluorescence values. The results showed that there were differences (P<0.01) between raw and reconstituted milk. The plot of values in each mixed level of raw and reconstituted milk had a correlation coefficient >0.97. The FAST method is a simple, rapid, low-cost and sensitive method enabling the detection of 5% reconstituted milk in fresh milk. The measurement of the Trp, AMP fluorescence values and calculation of the FAST index is a suitable method for large-scale monitoring of fresh milk samples. PMID- 16252346 TI - Liver fibrosis identification based on ultrasound images captured under varied imaging protocols. AB - Diagnostic ultrasound is a useful and noninvasive method in clinical medicine. Although due to its qualitative, subjective and experience-based nature, ultrasound image interpretation can be influenced by image conditions such as scanning frequency and machine settings. In this paper, a novel method is proposed to extract the liver features using the joint features of fractal dimension and the entropies of texture edge co-occurrence matrix based on ultrasound images, which is not sensitive to changes in emission frequency and gain. Then, Fisher linear classifier and support vector machine are employed to test a group of 99 in-vivo liver fibrosis images from 18 patients, as well as other 273 liver images from 18 normal human volunteers. PMID- 16252347 TI - Pretreatment of coking wastewater using anaerobic sequencing batch reactor (ASBR). AB - A laboratory-scale anaerobic sequencing batch reactor (ASBR) was used to pretreat coking wastewater. Inoculated anaerobic granular biomass was acclimated for 225 d to the coking wastewater, and then the biochemical methane potential (BMP) of the coking wastewater in the acclimated granular biomass was measured. At the same time, some fundamental technological factors, such as the filling time and the reacting time ratio (t(f)/t(r)), the mixing intensity and the intermittent mixing mode, that affect anaerobic pretreatment of coking wastewater with ASBR, were evaluated through orthogonal tests. The COD removal efficiency reached 38%-50% in the stable operation period with the organic loading rate of 0.37-0.54 kg COD/(m(3).d) at the optimum conditions of t(f)/t(r), the mixing intensity and the intermittent mixing mode. In addition, the biodegradability of coking wastewater distinctly increased after the pretreatment using ASBR. At the end of the experiment, the microorganism forms on the granulated sludge in the ASBR were observed using SEM (scanning electron microscope) and fluoroscope. The results showed that the dominant microorganism on the granular sludge was Methanosaeta instead of Methanosarcina dominated on the inoculated sludge. PMID- 16252348 TI - The impact of NiO on microstructure and electrical property of solid oxide fuel cell anode. AB - Ni-Ce(0.8)Sm(0.2)O(1.9) (Ni-SDC) cermet was selected as anode material for reduced temperature (800 degrees C) solid oxide fuel cells in this study. The influence of NiO powder fabrication methods for Ni-SDC cermets on the electrode performance was investigated so that the result obtained can be applied to make high-quality anode. Three kinds of NiO powder were synthesized with a fourth kind being available in the market. Four types of anode precursors were fabricated with these NiO powders and Ce(0.8)Sm(0.2)O(1.9) (SDC), and then were reduced to anode wafers for sequencing measurement. The electrical conductivity of the anodes was measured and the effect of microstructure was investigated. It was found that the anode electrical conductivity depends strongly on the NiO powder morphologies, microstructure of the cermet anode and particle sizes, which are decided by NiO powder preparation technique. The highest electrical conductivity is obtained for anode cermets with NiO powder synthesized by NiCO(3).2Ni(OH)(2).4H(2)O or Ni(NO(3))(2).6H(2)O decomposition technique. PMID- 16252349 TI - Dynamic Monte Carlo study on the probability distribution functions of tail-like polymer chain. AB - The configurational properties of tail-like polymer chains with one end attached to a flat surface are studied by using dynamic Monte Carlo technique. We find that the probability distribution of the free end in z direction P(R(z)) and the density profile rho(z) can be scaled approximately by a factor beta to be a length independent function for both random walking (RW) and self-avoiding walking (SAW) tail-like chains, where the factor beta is related to the mean square end-to-end distance . The scaled P(R(z)) of the SAW chain roughly overlaps that of the RW chain, but the scaled rho(z) of the SAW chain locates at smaller betaz than that of the RW chain. PMID- 16252350 TI - Influence of polarized bias and porous silicon morphology on the electrical behavior of Au-porous silicon contacts. AB - This paper reports the surface morphology and I-V curves of porous silicon (PS) samples and related devices. The observed fabrics on the PS surface were found to affect the electrical property of PS devices. When the devices were operated under different external bias (10 V or 3 V) for 10 min, their observed obvious differences in electrical properties may be due to the different control mechanisms in the Al/PS interface and PS matrix morphology. PMID- 16252351 TI - Life-extending hormone found in mice research may apply to all mammals. PMID- 16252352 TI - Kava, valerian no more effective than placebo. PMID- 16252353 TI - Hair dye doesn't up cancer risk. PMID- 16252354 TI - To lower your LDL further, add fiber. PMID- 16252355 TI - Do your heart a favor: eat fish. PMID- 16252356 TI - Chiropractic caveat. PMID- 16252357 TI - New age-related fitness formula for women. PMID- 16252359 TI - I had what my doctor called a mini-stroke. After an extensive evaluation from several other doctors, he concluded that I have a hole in my heart. He called it a PFO. He said I was born with it, but thought it should probably be closed now. What do you think? PMID- 16252358 TI - Results of vitamin E studies mixed. PMID- 16252360 TI - I get very sleepy in the late afternoon and again right after supper when watching television. Then I have trouble falling asleep or wake up early in the morning and can't get back to sleep. I am 77 and have no major health problems, except for being overweight. What would be helpful? PMID- 16252361 TI - The path to success. PMID- 16252362 TI - You say genomics, I say genetics... PMID- 16252363 TI - An academy of their own. PMID- 16252364 TI - [Recurrent laryngeal nerve paralysis after thyroid gland operations]. PMID- 16252365 TI - Giant mucinous cystadenoma of the appendix presenting with contralateral ilio femoral deep venous thrombosis (2005: 7b). PMID- 16252366 TI - Monitoring in polluted environments for integrated water-soil management. PMID- 16252367 TI - Another try. Nemzoff looking to start his own for-profit system. PMID- 16252368 TI - Part E draws support as a Medicare plan that covers all the bases. PMID- 16252369 TI - The neuropsychiatric burden of neurological diseases in the elderly. PMID- 16252370 TI - The concept of quality of life in dementia in the different stages of the disease. AB - In order to conceptually define quality of life (QOL) in dementia, the literature on QOL in the elderly population, in chronic disease and in dementia was studied. Dementia is a progressive, age-related, chronic condition and to avoid omissions within the dementia-specific concept of QOL, a broad orientation was the preferred approach in this literature study. Adaptation is a major outcome in studies investigating interventions aimed at improving QOL in chronic conditions, but to date, it has not been used in the definition of QOL. It is argued that adaptation is an important indication of QOL in people with chronic diseases and therefore also in dementia. Some crucial issues in assessing dementia-related QOL that are relevant to clarify the continuing debate on whether QOL, particularly in dementia, can be measured at all, are discussed. Then the following conceptual definition is offered: dementia-specific QOL is the multidimensional evaluation of the person-environment system of the individual, in terms of adaptation to the perceived consequences of the dementia. PMID- 16252371 TI - Lay perceptions about mental health: where is age and where is Alzheimer's disease? AB - Studies on laypersons' beliefs and knowledge about mental disorders have proliferated in recent years. However, attention has been focused mainly on depression and schizophrenia and on young adults. The aim of this paper is to summarize research in the area, and to discuss the need to expand research in the elderly population. The unique characteristics of older persons in terms of the prevalence and type of mental disorders, especially Alzheimer's disease (AD) and other dementias, as well as in terms of their being victims of "double jeopardy" require special attention and research. The present review has three main objectives. First, it summarizes the findings of studies examining different aspects of mental health literacy. Second, the importance of age in the study of mental health literacy is discussed. Third, findings of the few studies examining laypersons' beliefs in the area of AD are presented. Finally, research directions are suggested with special emphasis on the importance of geriatric mental health and mental health literacy. PMID- 16252372 TI - Attitudes to dementia and dementia care held by nursing staff in U.K. "non-EMI" care homes: what difference do they make? AB - BACKGROUND: There is doubt about the value of training in dementia care in U.K. nursing homes. We decided to estimate the association between nursing staff's attitudes to dementia and dementia care and their recognition of cognitive impairment in residents and other indicators of care practice in nonspecialist nursing homes derived from a probability sample of 445 residents in South-East England, and to relate this to previous training. METHODS: Prospective survey. The most senior nurse on duty was interviewed about each resident sampled, about their own training and experience, their attitude to restriction of egress and covert medication use, and asked to complete the Attitudes to Dementia Questionnaire (ADQ) and the dementia Care Styles Questionnaire (CSQ). Nurses were also asked about care practices in relation to restriction of egress and covert medication use in the home. Residents were interviewed using the Mini-mental State Examination (MMSE). RESULTS: One hundred and fifty-eight nurses were interviewed. Increased person-centered attitudes seem to be associated with better recognition of cognitive impairment independent of training and experience. The espousal of restrictive practices was also associated with better recognition, but only when analysis included nurses reporting on only one impaired resident. CONCLUSIONS: More person-centered attitudes are associated with better recognition of cognitive impairment, despite perverse U.K. regulatory incentives; the need for training and support in developing person-centered dementia care for staff in "non-Elderly Mentally Infirm" ("non-EMI") care homes is supported by these results. PMID- 16252373 TI - The Carers' Needs Assessment for Dementia (CNA-D): development, validity and reliability. AB - BACKGROUND: A variety of interventions are available to support the caregivers of dementia patients. For the purposes of service planning, we developed an instrument to assess the needs of these caregivers and to determine whether needs are met. The reliability and validity of this new instrument was also investigated. METHODS: The development of the Carers' Needs Assessment for Dementia (CNA-D), was based on in-depth interviews and a focus group. The combined inter-rater and test-retest reliability was investigated among 45 dementia caregivers. Correlations of the CNA-D with the Zarit Burden Inventory were used to analyze concurrent validity. Content validity was investigated by performing a separate survey among 40 caregivers and 40 professionals. RESULTS: The CNA-D is a semi-structured research interview including 18 problem areas. For each problem area, the CNA-D offers several possible interventions. The relevance of the problem areas and the interventions (content validity) was confirmed by most of the study participants. Significant positive associations were found between the total score of the Zarit Burden Inventory and the number of problems and the number of unmet needs according to the CNA-D. The agreement between the interviewers was "excellent" (kappa above 0.75) in 73.7% of the problem areas and in 69.9% of the interventions. CONCLUSIONS: The CNA-D is a valid and reliable instrument for comprehensively assessing the needs of dementia caregivers. PMID- 16252374 TI - The effects of the implementation of snoezelen on the quality of working life in psychogeriatric care. AB - BACKGROUND: Dementia among nursing home residents is often accompanied by high care dependency and behavioral disturbances, resulting in an increased workload for the caregivers. Snoezelen, integrated into 24-hour dementia care, is an approach that might improve the quality of working life of dementia caregivers. This study aims to investigate the effectiveness of integrated snoezelen on work related outcomes (workload and psychological outcomes) of caregivers in psychogeriatric nursing homes. METHODS: A quasi-experimental pre- and post-test design was used, comparing six psychogeriatric wards that implemented snoezelen in 24-hour care to six control wards that continued giving usual care. One hundred and twenty-nine Certified Nursing Assistants (CNAs) were included in the pre-test and 127 CNAs in the post-test. The six intervention wards received a 4 day in-house training program. The intervention further consisted of implementation activities on the ward (e.g. stimulus preference screening, workgroup), three in-house follow-up meetings and two general meetings. Measurements on workload, perceived problems, stress reactions, job satisfaction and burnout were performed at baseline and after 18 months. RESULTS: A significant treatment effect in favor of the experimental group was found for time pressure, perceived problems, stress reactions and emotional exhaustion. CNAs of the experimental group also improved on their overall job satisfaction score. In particular, they were more satisfied with the quality of care and with their contact with residents. CONCLUSION: The implementation of snoezelen improved the quality of the working life of dementia caregivers. PMID- 16252375 TI - Effect of antipsychotics on mortality in elderly patients with dementia: a 1-year prospective study in a nursing home. AB - BACKGROUND AND OBJECTIVES: The use of risperidone or olanzapine to treat behavioral problems associated with dementia is no longer recommended in the U.K. because of the increased risk of cerebrovascular adverse effects (CVAEs) and/or mortality. To evaluate the risks and benefits of antipsychotics, we measured the rate of mortality in patients with dementia, Alzheimer's disease (AD) and vascular/mixed dementia and compared mortality rates between those who had received antipsychotics and those who had not received antipsychotics. METHODS: A total of 273 subjects were assessed at baseline, 6 months and 12 months using a 1 year prospective follow-up design. Mortality rates between groups were compared using a Kaplan-Meier curve and log-rank statistics. Relative risks (RRs) were examined by the Cox proportional-hazards model. RESULTS: The overall 1-year mortality rate in dementia was 23.8%. The mortality rate in those who had not received antipsychotics (26.8%) was higher than that in those who had received antipsychotics (20.6%). RR and 95% confidence interval (CI) of mortality, when we compared those who had not received antipsychotics with those who had received antipsychotics, was 1.277 (95% CI 1.134-1.437) after controlling for age, severity of dementia, medical comorbidities, cognitive impairment (measured by the Korean version of the Mini-mental State Examination (MMSE)) and behavioral and psychological symptoms of dementia (BPSD), measured by the Behavioral Pathology in Alzheimer's Disease Rating Scale, Korean version (BEHAVE-AD-K). When those who had not received antipsychotics were compared with those who had received both risperidone and haloperidol, RR (95% CI) was 1.225 (1.101-1.364). CONCLUSION: This study does not support reports that antipsychotics increase mortality in dementia. PMID- 16252376 TI - The relationship between sleep disturbance and morale in Japanese elderly people. AB - BACKGROUND: Complaints of sleep disturbance are common in elderly individuals. The quality of life (QOL) for people who have insomnia is thought to be worse than for those who do not have insomnia. In this study we investigated the influence of disturbed sleep on morale in elderly people who live independently. METHODS: A survey of the necessity of public nursing care for all those aged over 65 years was performed in Kumamoto city, Japan. Three hundred subjects from the elderly population living at home without special care were sampled at random and they filled out a questionnaire regarding sleep, psychiatric symptoms and attitudes towards their own aging. RESULTS: A logistic regression analysis found psychiatric symptoms and problems keeping awake to be independently related to a negative attitude towards one's own aging. Neither sleep, sex nor age demonstrated any relationship with the negative attitudes of elderly individuals. CONCLUSION: Excessive daytime sleepiness is related to poor morale regardless of both the quality and quantity of sleep and psychiatric symptoms. PMID- 16252377 TI - Dementia of acute onset in the Canadian Study of Health and Aging. AB - BACKGROUND: Although most people with dementia experience an insidious onset of symptoms, in some cases onset can be acute. The importance of acute onset is unclear. Some reports suggest that it portends a worse course. METHODS: We performed a secondary analysis of the clinical examination cohort (n=2914) of the Canadian Study of Health and Aging (CSHA). We defined "acute onset of dementia" from the Cambridge Examination for Mental Disorders in the Elderly (CAMDEX) questionnaire, conducted with an informant. People with dementia of acute onset were compared to those with dementia of insidious onset for development of adverse outcomes of death and institutionalization over 5 years. RESULTS: Of the 1132 people who had dementia, 130 (11.5%) met criteria for acute onset. Compared with gradual-onset dementia patients, those with acute-onset dementia were more often men (42% vs. 30%, p < 0.05), resided in nursing homes (75% vs. 63%, p< 0.05), had vascular risk factors (72% vs. 47%, p < 0.05), and a Hachinski Ischemia Scale (HIS) score > or = 7 (64% vs. 19%, p < 0.05). More patients with dementia of acute onset than gradual onset were diagnosed with vascular dementia (55% vs. 13%; p < 0.05). Adjusted hazard ratios (HRs) for survival and institutionalization in the acute-onset group were 0.93 [95% confidence interval (CI) 0.7-1.2] and 0.76 (95% CI 0.4-1.3), respectively, compared with the gradual onset group. CONCLUSIONS: People with acute-onset dementia had more vascular risk factors than those with gradual-onset dementia across all dementia diagnoses, and lower risks of institutionalization but worse survival. Routine inquiry about the onset of dementia might help to better clarify prognoses in patients with dementia. PMID- 16252378 TI - Instrumental activities of daily living scale for dementia screening in elderly people. AB - OBJECTIVE: To develop and validate an Instrumental Activities of Daily Living Scale for elderly people (IADL-E) to use in conjunction with cognitive screening tests for dementia in an educationally and socioculturally heterogeneous population. METHOD: Eleven IADL items were selected and weighted for major factors causing heterogeneity in the population--gender, education, social (rural/urban) setting and age. Each item was rated for its applicability (yes/no), degree of disability (scored from 0 to 2) and causative impairment (cognitive and/or physical). From this a composite index of cognitive (CDI) or physical (PDI) disability was derived. Validation was performed retrospectively on 240 subjects: 135 without and 105 with dementia by DSM-IV. RESULTS: The IADL-E had a high internal consistency (alpha = 0.95). The area under the receiver operating characteristic (ROC) curve was 0.97 (CI = 0.94-0.99). A cutoff score of 16 on CDI provided a sensitivity of 0.91, specificity 0.99 and positive predictive value 0.76 (at 5% base rate). IADL-E correlated highly with clinical (DSM-IV, kappa = 0.89), functional (CDR, 0.82) and cognitive (Mini-mental Status Examination, MMSE, 0.74) diagnoses. It showed good responsiveness, with the change on CDI over a median of 23 months correlating significantly with that on MMSE (coefficient = -0.382, CI = -0.667 to -0.098; p=0.009). Individual items had good interrater and test-retest reliability. CONCLUSIONS: The IADL-E is a reliable, sensitive and responsive scale of functional abilities useful in dementia screening in a socioculturally heterogeneous population. PMID- 16252379 TI - A randomized trial of early psychiatric intervention in residential care: impact on health outcomes. AB - BACKGROUND: The prevalence of psychological and behavioral disturbances among older adults living in residential care facilities is high, and it has been shown previously that people with such symptoms have poorer health outcomes. This study was designed to assess the efficacy of an early psychiatric intervention on the 12-month health outcomes of older adults admitted to residential care facilities in Perth, Western Australia. We hypothesized that subjects in the intervention group would have better mental and physical health outcomes than controls. METHODS: The study was designed as a randomized, single-blinded, controlled trial. All subjects aged 65 years or over admitted to one of the 22/26 participating residential care facilities of the Inner City area of Perth were approached to join the study and were allocated randomly to the intervention or usual care group. Demographic and clinical information (including medications and use of physical restraint) was gathered systematically from all participants at baseline, and at 6 and 12 months. At each assessment, the Geriatric Depression Scale (GDS), the Health of the Nation Outcome Scales for older adults (HoNOS 65+), the Mini-mental State Examination (MMSE) and the Neuropsychiatric Inventory (NPI) were administered. Subjects in the intervention group who screened positive at the baseline assessment for psychiatric morbidity were reviewed within a 2 week period by the Inner City Mental Health Service of Older Adults (ICMHSOA). If clinically appropriate, mental health services were introduced without the involvement of the research team. RESULTS: One hundred and six subjects and their next of kin consented to participate in the study (53 in each group). Mental health screening and early referral to a psychogeriatric service did not significantly change the average number of medical contacts, self-rated health, use of psychotropic or PRN medication, use of physical restraint, 12-month mortality, or mental health outcomes, as measured by the GDS-15, HoNOS 65+ and NPI (p > 0.05 for all relevant outcomes). CONCLUSION: Systematic mental health screening of older adults admitted to residential care facilities and early clinical intervention does not change 12-month health outcomes. More effective interventions to improve the health outcomes of older adults with psychological and behavioral disturbances admitted to residential care facilities are needed. PMID- 16252381 TI - What do subjective cognitive complaints in persons with aging-associated cognitive decline reflect? AB - BACKGROUND: Subjective cognitive complaints have been included in diagnostic concepts such as Aging-Associated Cognitive Decline (AACD) aiming to identify older adults with cognitive impairments at high risk of developing dementia. Although several studies in normal aging have found that subjective cognitive complaints are related to depressive affect and personality factors, little is known as to whether this is also true for older adults with AACD. METHODS: In 123 older adults diagnosed with AACD and 291 controls, the role of actual cognitive performance, depressive affect, neuroticism and conscientiousness in predicting subjective cognitive complaints was investigated. In separate ordinary least squares regression analyses for both groups with gender, age, years of schooling, cognitive performance, depressive affect, neuroticism and conscientiousness as predicting variables, in the control participants, gender, age, depressive affect and neuroticism were related to subjective cognitive complaints, whereas in the AACD participants only gender and neuroticism accounted for variance in subjective cognitive complaints. Testing for group differences in predictive power, revealed differential effects for gender, depressive affect and neuroticism. CONCLUSIONS: As subjective cognitive complaints in the AACD group were related to neuroticism and gender rather than to cognitive performance, their inclusion in diagnostic concepts such as AACD should be revaluated. However, the nature of subjective cognitive complaints might be qualitatively different in persons diagnosed with AACD compared to those stated by normal older adults. PMID- 16252380 TI - A double-blind, randomized clinical trial to assess the augmentation with nimodipine of antidepressant therapy in the treatment of "vascular depression". AB - BACKGROUND: Cerebrovascular disease may cause "vascular depression" (VaD). Calcium channel-blockers are presumed treatments for cerebrovascular disease and might be expected to improve depression and prevent recurrence. OBJECTIVE: To examine the efficacy and tolerability of the use of nimodipine as an augmentation of fluoxetine in the treatment of VaD. DESIGN: A double-blind, randomized clinical trial in which 101 patients with VaD (Alexopoulos criteria) were treated with fluoxetine at standard doses. Patients were randomized to placebo (n=51) or nimodipine (n=50). Treatment outcomes were assessed using the Hamilton Depression Rating Scale (HDRS) regularly up to 8 months after treatment initiation. RESULTS: Depression was reduced in 63% of patients, but those whose treatment was enhanced with nimodipine had greater improvements overall by repeated measures analysis of covariance (ANCOVA) (F(1.80) = 9.76, p=0.001). In addition, a greater proportion of patients treated with fluoxetine-nimodipine (54% vs. 27%) exhibited full remission (chi2(d.f. 1)= 7.3, p = 0.006), with the number needed to treat (NNT) equal to 4 (95% CI 2-12). Of those experiencing full remission in the first 61 days, fewer patients on fluoxetine-nimodipine (3.7%) developed recurrence of major depression as compared to those on fluoxetine alone (35.7%) (chi2(d.f. 1) = 7.56, p = 0.006), NNT 3 (95% CI 2-9). Side-effects were noted in 33.3% of patients in the control group and 48% of the experimental group (chi2(d.f. 1) = 2.25, p = 0.133). CONCLUSIONS: In treating VaD, augmentation of fluoxetine with nimodipine led to better treatment results and lower rates of recurrence. These findings support the argument that augmentation of antidepressant therapy might be helpful in the treatment of cerebrovascular disease, which is involved in the pathogenesis of this type of depression. PMID- 16252382 TI - Is there a need to study behavioral and psychological signs and symptoms of dementia across cultures? PMID- 16252384 TI - [The EEG correlates of delayed mental development in adolescents]. AB - Fifty-eight male adolescents aged 15-17 years have been divided into 3 groups: 31 patients with mental infantilism syndrome were included in the study group, 14 with organic brain disorders--in the comparison group and 13 psychiatrically and neurologically normal subjects--in the control group. EEG was recorded from 12 leads monopolarly at rest, during hemisphere-specific cognitive tasks performance and exposure to aversive sound stimulation. Two-Hz wide EEG spectral ranges--A, theta1, theta2, alpha1 and alpha2--were analyzed. Only in the study group, there were decreased values of alpha2 spectral power (SP) and reduced reactivity to functional tests and insufficient lateral differentiation of the reactions during the cognitive tasks performance. Maximal deviation of the SP reactivity indices as compared to the comparison group was found in alpha2 range. The disturbances obtained indicate the delay of brain maturation and retardation in formation of specific neural networks. The results of the study demonstrate that indices of alpha2 SP may be considered as objective criteria of delayed mental maturation. PMID- 16252385 TI - [Time-organization of EEG patterns' structure in anxiety and phobic disorders]. AB - Thirty-five patients, aged 19-48 years (mean age 38 years) with anxiety and phobic disorders were examined. According to ICD-10 criteria--social phobia (F40.1), panic disorder (F41.0), somatoform autonomic dysfunction (F45.3) were diagnosed. Using electroencephalography data, qualitative and quantitative characteristics of the time- and spatial-organization of brain EEG activity in anxiety and phobic disorders of different severity were established. It were determined 4 types of wave interactions between EEG components, which reflected a different extent of the regulatory mechanisms lesions: 2 structures with one core component (alpha or beta), a structure with two core components and a non organized structure. PMID- 16252383 TI - [Autonomic cardiovascular regulation in patients with tics and Tourette syndrome]. AB - Autonomic cardiovascular regulation has been assessed in patients aged 4-15 years with Tourette syndrome (n = 22) and other tic disorders (n = 48). Symptom significance was estimated by a number of hyperkinetic episodes per 20 minutes, tic scale and variants of the disease course. The functional condition of autonomic nervous system was studied clinically and using spectral analysis of heart rate variability in both upright and supine positions. Negative correlation between the ratio of sympathetic and vagus influences and severity of the disease was found: the severer were tic symptoms, the stronger was a trend to vagotonia (beta = -0.36; p < 0.0025; F > 4.0). In orthostatic test, patients with Tourette syndrome demonstrated an unfavorable hypersympathicotonic type of cardiovascular system reaction. Patients were treated during 4 weeks with glycinum (0.2 +/- 0.1 mg/day), phenibutum (0.5 +/- 0.25 mg/day), clonazepam (1.5 +/- 0.5 mg/day), tiapride (200 +/- 100 mg/day), haloperidol (1-1.5 mg/day), rispolept (2 mg/day). There was no negative effect of the drugs on heart rate variability. On the contrary, the therapy reduced hyperkinetic symptoms and corrected autonomic influences on the sinus rhythm. It is suggested that changes in autonomic cardiovascular regulation might be of secondary character and do not need any special correction. PMID- 16252386 TI - [Dopamine receptor DRD4 gene polymorphism and its association with schizophrenia spectrum disorders and personality traits of patients]. AB - A vast body of associative studies reported a role of highly polymorphic dopamine receptor DRD4 gene in regulation of emotional processes and development of mental disorders. The present study addresses allele, genotype and haplotype distribution of 3 polymorphic DRD4 markers (-809G/A, -616G/C N -521C/T) in Russian patients with schizophrenia spectrum disorders and their relation to the disease and personality traits. A sample included 151 patients with iCD-10 diagnosis of schizophrenia, schizoaffective psychosis and schizotypal personality disorders, 89 their first-degree non-psychotic relatives and 131 mentally healthy individuals. No differences in allele and genotype frequency was found between the patients and the controls. Transmission disiquilibrium test (TDT) did not reveal a preferential transmission of either allele from parents to proband. The 521C/T N -616G/C markers were linked to the disease when the EH program has been used in the analysis. Patients with the GG (-809G/A) and GG (-616G/C) genotypes had higher scores on the Hypomania scale (MMPI) comparing to the GA(-809G/A)+AA( 809G/A) and GC(-616G/C)+CC(-616G/C) genotypes but the association did not reach a level of significance (p = 0.06). The results confirmed the literature reports on the relation of the DRD4 gene to schizophrenia and personality traits related to social activity. PMID- 16252387 TI - [Cognitive disturbances and attempts of their correction in multiple sclerosis]. PMID- 16252388 TI - First whooping cough vaccine for adolescents. PMID- 16252389 TI - New drug extends lives of brain cancer patients. PMID- 16252390 TI - First DNA-based test to detect cystic fibrosis. PMID- 16252391 TI - Sealant after brain surgery. PMID- 16252392 TI - First-of-a-kind device treats aneurysms. PMID- 16252393 TI - Experimental shingles vaccine proves effective in nationwide study. PMID- 16252394 TI - Study shows programs can teach children to eat healthier. PMID- 16252395 TI - Does sex make a difference? PMID- 16252396 TI - Computer-assisted surgery: an update. PMID- 16252397 TI - Antiperspirant awareness: it's mostly no sweat. PMID- 16252398 TI - Reducing the risk of rabies. PMID- 16252401 TI - A new era of hope for people with lupus. PMID- 16252400 TI - Drug name confusion: preventing medication errors. PMID- 16252402 TI - [The 78th congress of the Japanese Biochemical Society. Kobe, Japan. October 19 22, 2005. Abstracts]. PMID- 16252399 TI - Battling lupus. PMID- 16252403 TI - [One-stage plasty of esophagus in extravasal compression of celiac trunk]. PMID- 16252404 TI - [Successful conservative treatment of chylopericardium]. PMID- 16252405 TI - [Differential diagnosis of diseases associated with jaundice]. PMID- 16252406 TI - [Venous system of penis]. PMID- 16252407 TI - [Role of staged reconstruction of anterior abdominal wall in large and gigantic postoperative hernias]. PMID- 16252408 TI - [Role of plants: confined to nutrition and therapy. Proceedings of a conference, 7 May 2004, Rome, Italy]. PMID- 16252409 TI - The Communicative Cardiac Cell. Proceedings of a workshop, January 15-19, 2005, Sintra, Portugal. PMID- 16252411 TI - [36th National Gastroenterology, Hepatology and Endoscopy Symposium]. PMID- 16252410 TI - Summary of the California blue ribbon panel report on anesthesia. AB - At the request of the Dental Board of California, a panel reviewed mortality data from the Dental Board, lawsuits from a major California malpractice insurance company, anesthesia regulations from other states, and the published scientific literature. In California between 1991 and 2000, there were 12 deaths related to general anesthesia permits, 0 deaths related to conscious sedation permits, and 8 deaths related to nonpermit holders (four deaths with oral sedation in children and four deaths with local anesthesia alone). The panel was concerned about the increased use of repeated oral or sublingual doses of sedatives and recommended a certificate process. The panel recommended a standing committee to access significant anesthesia/sedation-related misadventures and to determine how such mishaps could be prevented. The data reviewed and recommendations made are summarized in this report. PMID- 16252412 TI - Where a lump on the back proves to be a warfarin-induced hematoma. PMID- 16252413 TI - Where an unusual lesion is treated aggressively. PMID- 16252414 TI - Benign breast disease and breast cancer. PMID- 16252415 TI - Atorvastatin in patients with type 2 diabetes mellitus undergoing dialysis. PMID- 16252416 TI - Atorvastatin in patients with type 2 diabetes mellitus undergoing dialysis. PMID- 16252417 TI - Quality of care in U.S. hospitals. PMID- 16252418 TI - Quality of care in U.S. hospitals. PMID- 16252419 TI - Diagnosis from the blood smear. PMID- 16252420 TI - Update: West Nile virus activity--United States, 2005. AB - This report summarizes West Nile virus (WNV) surveillance data reported to CDC through ArboNET as of 3 a.m. Mountain Daylight Time, October 25, 2005. PMID- 16252421 TI - Abstracts from the joint annual meeting of the American Epilepsy Society and the American Clinical Neurophysiology Society. Washington DC, USA, December 2-6, 2005. PMID- 16252422 TI - TCT 2005. Abstracts of the 17th Annual Transcatheter Cardiovascular Therapeutics Symposium, October 17-21, 2005, Washington DC, USA. PMID- 16252424 TI - Abstracts of the XLth International Symposium on the Spontaneously Hypertensive Rat and Cardiovascular Risk. September 17-18, 2004, Osaka, Japan. PMID- 16252423 TI - Abstracts of the 3rd European Meeting on Vascular Biology and Medicine, Hamburg, Germany, September 28-30, 2005. PMID- 16252425 TI - Abstracts of the 28th European Conference on Visual Perception, A Coruna, Spain, 22-26 August 2005. PMID- 16252426 TI - Abstracts of the 58th Annual Meeting of the Japanese Association for Thoracic Surgery, Okayama, Japan, October 5-7, 2005. PMID- 16252428 TI - HUPO 2nd and IUBMB XIX World Congress, October 8-11, 2003, Montreal, Canada. Abstracts. PMID- 16252427 TI - EVER 2005. European Association for Vision and Eye Research. October 5-8, 2005, Vilamoura, Portugal. Abstracts. PMID- 16252429 TI - Abstracts of the 36th Annual Meeting of the Austrian Society of Internal Medicine, Salzburg, 29 September-1 October 2005. PMID- 16252431 TI - NoiseCon 2005 and the 150th Meeting of the Acoustical Society of America. Minneapolis, Minnesota, USA, October 17-19, 2005. Abstracts. PMID- 16252430 TI - [Abstracts of the IVth Congress of the French Society of Vascular Medicine, Bordeaux, France, 22-24 September 2005]. PMID- 16252432 TI - The role of health education and behavior in public health genetics. AB - This article highlights the important role of health behavior and health education (HBHE) research in public health genetics. Broadly defined, public health genetics is the integration of genetic advancements and technologies in the study and practice of public health. The potential role of HBHE within this area is presented across two intersecting continua--namely, the continuum between research and practice and the continuum between individual/personalized medicine and population health. The authors begin this article with an overview of current issues arising from the use of genetic information to improve the public's health and provide a framework for understanding the multidimensional role of HBHE research in translating genetic research into medical and public health practice. An introduction to the nine articles and two practice notes included in this special issue is also provided to draw attention to the crosscutting themes and issues presented. PMID- 16252433 TI - Value of oxaliplatin treatment in heavily pretreated patients with non-Hodgkin's lymphoma. AB - In order to assess the efficacy and toxicity profile of oxaliplatin, a third generation platinum derivate active against several solid tumors, we carried out a study in a group of heavily pretreated patients with non-Hodgkin's lymphoma (NHL). Between August 2003 and May 2004, 19 pretreated patients were enrolled in a phase II trial and were treated with oxaliplatin. The drug was administered intravenously on day 1 of a 21-day schedule, at a dose of 130 mg/m2 for a total of 6 cycles. One (5%) patient achieved complete remission (CR) and 5 patients (27%) had partial response (PR), thus giving an overall response rate of 32%. The patient in CR suffered from an aggressive B NHL. One of the 5 patients in PR had an aggressive B NHL, whereas the remaining 4 had an indolent B NHL. The treatment was well tolerated with minimal hematologic and extrahematologic toxicity. These data suggest and confirm the efficacy and low toxicity of oxaliplatin in the treatment of patients with heavily pretreated NHL. Further trials using oxaliplatin alone or in combination with other conventional drugs are needed. PMID- 16252434 TI - [XXIIth Congress of the French Society of Endocrinology, Strasbourg, France, 9-12 October 2005. Abstracts]. PMID- 16252435 TI - The detection of malingering in memory performance: the sensitivity and specificity of four measures in a UK population. AB - OBJECTIVES: To investigate the validity of a clinical neuropsychological battery for the detection of malingering on tests of memory. METHODS: A simulated scenario design was developed to investigate the effectiveness of a battery of four neuropsychological tests in the detection of malingering; the Coin in the Hand Test (CIH), Autobiographical Memory Index (AMI), Rey I 5-Item Test (RIT),and the Wechsler Mental Control Test (MCT). The performances of patients with an acquired brain injury (N = 40) were compared with two groups of controls instructed either to simulate a head injury performance (N = 40) or do their best (N = 40). RESULTS: The CIH and MCT demonstrated good validity and displayed high sensitivity and specificity. The RIT and the AMI was relatively poor in distinguishing between simulators and patients. CONCLUSIONS: The sensitivity and specificity of all four tests to the detection of malingering has been assessed. Two of the tests the CIH and MCT would be useful as a quick and accurate screening tool for detecting malingering. PMID- 16252436 TI - Does exposure to analgesics in utero cause schizophrenia? PMID- 16252437 TI - Emotional development, shame, and adaptation to child maltreatment. PMID- 16252438 TI - Methadone--an old analgesic with new tricks. PMID- 16252439 TI - Abstracts of the XII European Pediatric Rheumatology Congress and annual scientific meeting of PReS, 15-18 September 2005, Versailles, France. PMID- 16252440 TI - Objective assessment of epidural anesthesia using epidural catheter-derived evoked potentials. PMID- 16252441 TI - Preliminary exploration of burning mouth and burning feet: Is there a common etiology? PMID- 16252442 TI - Abstracts of the 2nd COST B22 Congress on Drug Discovery and Development for Parasitic Diseases, Siena, Italy, 29 September-1 October 2005. PMID- 16252443 TI - Applied superconductivity and superfluidity for the exploration of the Moon and Mars. AB - We discuss how superconductivity and superfluidity can be applied to solve the challenges in the exploration of the Moon and Mars. High sensitivity instruments using phenomena of superconductivity and superfluidity can potentially make significant contributions to the fields of navigation, automation, habitation, and resource location. Using the quantum nature of superconductivity, lightweight and very sensitive diagnostic tools can be made to monitor the health of astronauts. Moreover, the Moon and Mars offer a unique environment for scientific exploration. We also discuss how powerful superconducting instruments may enable scientists to seek answers to several profound questions about nature. These answers will not only deepen our appreciation of the universe, they may also open the door to paradigm-shifting technologies. PMID- 16252444 TI - Depressed youth, suicidality and antidepressants. Comment. PMID- 16252446 TI - Universal varicella vaccination. Comment. PMID- 16252445 TI - Magnetic levitation-based Martian and Lunar gravity simulator. AB - Missions to Mars will subject living specimens to a range of low gravity environments. Deleterious biological effects of prolonged exposure to Martian gravity (0.38 g), Lunar gravity (0.17 g), and microgravity are expected, but the mechanisms involved and potential for remedies are unknown. We are proposing the development of a facility that provides a simulated Martian and Lunar gravity environment for experiments on biological systems in a well controlled laboratory setting. The magnetic adjustable gravity simulator will employ intense, inhomogeneous magnetic fields to exert magnetic body forces on a specimen that oppose the body force of gravity. By adjusting the magnetic field, it is possible to continuously adjust the total body force acting on a specimen. The simulator system considered consists of a superconducting solenoid with a room temperature bore sufficiently large to accommodate small whole organisms, cell cultures, and gravity sensitive bio-molecular solutions. It will have good optical access so that the organisms can be viewed in situ. This facility will be valuable for experimental observations and public demonstrations of systems in simulated reduced gravity. PMID- 16252447 TI - [The system spared from exhaustion]. PMID- 16252448 TI - [Clinical ethical advice gains importance]. PMID- 16252449 TI - [Geriatric nurses: the daily passing of time]. PMID- 16252450 TI - [The burden of aging. A society of erroneous utility]. PMID- 16252452 TI - [Hospital nursing beds are much in need]. PMID- 16252451 TI - [Twice the moral self-knowledge]. PMID- 16252453 TI - [Norovirus in the forefront]. PMID- 16252454 TI - [Delegation of medical problems]. PMID- 16252455 TI - [The attitude of the Society of Social Justice on the administration of medications]. PMID- 16252456 TI - [The 2005 health prize for to four clinics from the Administration for Knowledge and Health ]. PMID- 16252457 TI - [5th International Conference for Nurses and Nursing]. PMID- 16252458 TI - [This article does not again mirror the knowledge of nurses]. PMID- 16252459 TI - [Book title: the mood-maker for nurses]. PMID- 16252460 TI - [Patients need influential nurses]. PMID- 16252461 TI - [Electronic competition by E-mail]. PMID- 16252462 TI - [Industrial leadership]. PMID- 16252463 TI - [Mullein helps in hoarseness]. PMID- 16252464 TI - Counting swabs. PMID- 16252465 TI - You find retention a real problem? PMID- 16252466 TI - New procedures in the recovery unit. PMID- 16252467 TI - Awareness during anaesthesia: what can be done to prevent it? AB - Awareness during general anaesthesia is a potentially traumatising risk of any general anaesthetic, which can have lasting effects on the patients who experience it. This article assesses the issues and causes of anaesthetic awareness, together with the current status of research being conducted into its prevention, and the effect of market forces and litigation. PMID- 16252468 TI - The National Patient Safety Agency and theatre practitioners. AB - Theatre practitioners have been involved in patient safety for years. Each time you check a patient into the department you are using a risk management tool: the check-list. Each time you check the position of a patient on the operating table, check their consent form, ensure the sterile field is maintained and that the instrument count is correct, you are intrinsically involved in patient safety. PMID- 16252469 TI - Preoperative visiting: landmarks of the journey. AB - Our fifteen bed, five day recovery unit accepts adult and paediatric patients and serves seven theatres dedicated to eye and ENT surgery. The aim of this article is to provide an insight into the practicalities involved in introducing a preoperative visiting programme and associated leaflet. These initiatives aim to give continuity of care, reduce stress and anxiety, and promote patients' coping strategies, by increasing the patients' knowledge and understanding of their perioperative journey (Malin & Teasdale 1991). PMID- 16252470 TI - Prewarming: preventing intraoperative hypothermia. AB - Perioperative hypothermia can be followed by severe complications. The greatest proportion of temperature decrease is attributed to heat redistribution, which mainly occurs during the first hour of anaesthesia and is difficult to treat intraoperatively. Prewarming, based on active warming techniques, has been proposed. Even a short period of prewarming may significantly increase peripheral tissue temperature, minimise normal core-to-peripheral temperature gradient, and keep core temperature within normal limits. PMID- 16252471 TI - Papers from the COST Action 857 Workshop V Biodiversity and population genetics of the Apicomplexa (Congress Centre of the Bambino Gesu Hospital, Rome, November 8-9, 2004). PMID- 16252472 TI - Molecular epidemiology of human cryptosporidiosis. AB - Species within the genus Cryptosporidium are protozoan parasites that infect a wide range of vertebrates, and represent a significant cause of morbidity and mortality in those animals. In humans, cryptosporidiosis is a common cause of diarrhoeal disease with a global distribution. Unravelling the epidemiology of human infection has proven to be difficult, due to the existence of multiple transmission routes (person-to-person, animal-to-person, waterborne, foodborne and airborne transmission), and to the difficulties in identifying the different species using conventional criteria, such as oocyst morphology. The advent of molecular techniques has had a remarkable impact on the way the epidemiology of cryptosporidiosis can be studied. Molecular investigations have shown that the vast majority of human cases are caused by C. hominis and C. parvum. Interestingly, differences in geographical and temporal distribution, disease presentations and risk factors for infection have been identified for both C. hominis and C. parvum. Further, molecular analyses have revealed that other species, including C. meleagridis, C. felis, C. canis, C. suis, C. muris and two Cryptosporidium genotypes, can infect humans and may be linked to clinical disease, not only in immunocompromised but also in immunocompetent individuals. PMID- 16252473 TI - Cryptosporidium and cryptosporidiosis in Denmark--current status. AB - The genus Cryptosporidium comprises a group of protozoan parasites that infect a broad variety of vertebrates causing severe diarrhoeal illness in immunocompromised as well as immunocompetent hosts. Although molecular heterogeneity of the genus is being increasingly recognised, traditional diagnostic methods do not discriminate all species/subtypes, and population genetic studies of these parasites, using discriminatory molecular markers, have only been published recently. In Denmark, Cryptosporidium research has focussed mainly on detection methods, pathogenicity and veterinary aspects. The present paper gives an overview of recent and ongoing Cryptosporidium research in Denmark with an emphasis on molecular approaches to study epidemiology and transmission. PMID- 16252474 TI - Molecular approaches to monitor parasite genetic complexity in the transmission of Plasmodium falciparum malaria. AB - Determination of the number of parasite clones present in a malaria infection is usually based on Polymerase Chain Reaction (PCR) amplification of Plasmodium polymorphic genomic sequences from peripheral blood. This method however does not provide information on the developmental stages of the parasites detected, or on the potential trasmissibility of the detected genotypes to the Anopheles vector. Reverse Transcriptase-PCR assays on P. falciparum mRNAs produced specifically in sexual stages have been developed in the past few years in order to detect and genotype circulating gametocytes, the parasite transmission stages, and are discussed in this review. Assays based on P. falciparum gamete-specific gene pfs25 and gametocyte-specific polymorphic gene pfg377 can detect presence of subpatent gametocytes in infected blood, can identify the pfg377 allele(s) specifically carried by the sexual stages, and detect coexistance of gametocytes of different genotypes. These assay have been used for the first time in field studies in a region of Sudan where malaria is seasonal, and they characterised parasite clonality and pattern of gametocyte production in the subpatent parasitaemias observed in the long malaria-free season. The method of specifically detecting and genotyping gametocytes in natural infections is proving to be a useful tool in investigating parasite transmission dynamics in field studies. This approach can be further improved by developing a multilocus RT-PCR assay which includes additional polymorphic gametocyte-specific transcripts. Candidate genes can be identified from the available data on the P. falciparum genome sequence and from recent analyses of parasite stage-specific transcriptomes. PMID- 16252475 TI - How old are the extant lineages of Toxoplasma gondii? AB - Most known isolates of Toxoplasma gondii belong to one of only three lineages, which are presumed to be clonal. Three models have been proposed for the evolutionary relationship of these lineages to the other extant lineages: Model (a) proposing that all lineages are derived from a most recent common ancestor (MRCA) in the distant past, Model (b) that all lineages are derived from a MRCA in the very recent past, and Model (c) that the clonal lineages share a recent MRCA but are related to the other lineages only in the distant past. Here, I test these models using DNA intron and coding-sequence data for loci at 14 genes, using three different methods to calculate the time of the MRCA. All of the calculations agree that the MRCA of the clonal lineages was > 70% of the age of the MRCA of all lineages, thus favouring Model (a). The MRCA may have existed approximately 150,000 years ago, with the clonal lineages expanding in prevalence approximately 10,000 years ago. PMID- 16252476 TI - The unusual architecture and predicted function of the mitochondrion organelle in Cryptosporidium parvum and hominis species: the strong paradigm of the structure function relationship. AB - Cryptosporidium spp. is a protozoan parasite that causes widespread diarrhoeal disease in humans and other animals and is responsible for large waterborne outbreaks of cryptosporidiosis. Unlike many organisms belonging to the phylum Apicomplexa, such as Plasmodium spp. and Toxoplasma gondii, there is no clinically proven drug treatment against this parasite. Some aspects of the basic biology of Cryptosporidium spp. such as the understanding of key metabolic pathways or the full description of the organellar compartment are still lacking. Here I present evidence of the anomalous shape and substructure of the mitochondrion organelle in C. parvum and C. hominis, which is closer to the Guillardia theta nucleomorph structure rather than to the canonical mitochondrion of the proximate apicomplexan T gondii. The atypical architecture is accomplished by an altered organellar metabolone, inferred by in silico conceptual prediction and characterized by unusual, partial and/or reduced pathways. However, phylogeneticanalyses of the mitochondrion and mitochondrion-related loci hsp60, hsp70 (dnaK), alternative oxidase (AOX) and superoxide dismutase (SOD) in C. parvum show diversiform evolutionary pathways, suggesting a "chimera" derived organelle. Taken together these data depict peculiar and intriguing aspects of the C. parvum and C. hominis anomalous mitochondrion framework for further comparative analysis of the organelle within the Cryptosporidium spp. order. PMID- 16252477 TI - Prevalence, behavioural and social factors associated with Schistosoma intercalatum and geohelminth infections in Sao Tome and Principe. AB - A pilot study was conducted in schoolchildren from three main districts of Sao Tome to assess the relationship between the prevalence of infections caused by Schistosoma intercalatum or intestinal helminths and individual behaviour and social conditions. Coprological examination revealed an increase of schistosome infections and a persisting high endemicity for ascariasis and trichuriasis. Infection rates were 36.2% for S. intercalatum and 70.8%, 68.5% and 4.6% for Ascaris lumbricoides, Trichuris trichiura and Ancylostomidae, respectively. Out of the 47 children positive for S. intercalatum, 35 (74.5%) were co-infected with one or more geohelminths. Logistic regression analysis of data collected through questionnaire demonstrate that behaviour and/or social conditions in the house were positively associated with S. intercalatum or T. trichiura. Neither sex nor age groups were associated with infections, suggesting that low personal hygiene and sanitation practices were similar for all groups of children. These data are in accordance to those of other studies and highlight the importance of assessing multivariate factors that may contribute to the transmission of these diseases, in order to design integrated control approaches for schistosomiasis and geohelminthiasis which could have more rapid effects on reduction of infections as well as greater cost-effectiveness. PMID- 16252478 TI - Seroprevalence of acquired toxoplasmosis in HIV-infected and apparently healthy individuals in Jos, Nigeria. AB - Toxoplasma gondii IgG antibody seroprevalence was studied in two different populations of 219 HIV-infected patients and 144 apparently healthy individuals (AHIs). Clinical toxoplasmosis was assessed among the HIV-infected patients. Antibodies to T. gondii were detected in 85 (38.8%, 95% CI: 32.36%-45.26%) of the HIV-infected patients and in 30 (20.8%, 95% CI: 14.20%-27.46%) of the AHIs. Among the AIHs, males represented 22.0% of infections compared to females (20.0%) and individuals within age group 21-30 years accounted for the highest prevalence of 33.3% (95% CI: 11.56%-55.10%). There was no significant difference in the trend (Chi-square, P < or = 0.05). Assessment of epidemiological factors showed higher seroprevalence of Toxoplasma antibodies among those who eat rodents (29.6%) and those who constantly have contact with the soil (21.2%). Among the HIV-infected, individuals 31-40-years-old had the highest T. gondii seroprevalence (36.5%). Evaluation of the clinical findings of patients with concomitant toxoplasmosis and HIV infection greatly implicated fever (63.5%), headache (44.7%), rashes (41.2%) and anorexia (34.1%). This study contributes to the development of guidelines for the prevention and management of toxoplasmosis in HIV-infected patients and in apparently healthy individuals in a resource scarce setting. PMID- 16252479 TI - Morphometric discriminant analysis for the classification of Diplectanum (Monogenea: monopisthocotylea), parasites of Sphyraena flavicauda. AB - The authors have examined the morphology of Diplectanum cazauxi and Diplectanum bauchotae through the observation of 23 specimens (10 of D. cazauxi and 13 of D. bauchotae) found during May 2003 on the gills of 2 specimens of Sphyraena flavicauda Ruppell, 1838 collected at Ras Mohammed National Park, Egypt. A discriminant analysis was performed on the morphometric data of opisthaptor to determine which parameters more accurately distinguish the two species. The best discriminant parameters resulted to be the copulatory apparatus and the central bar, which determined the absence of misclassification in the model. The discriminant functions are reported for the two species. PMID- 16252480 TI - Survey on helminthofauna in pheasants from Eastern Europe. AB - In order to assess the parasitological status in imported pheasants, 51 birds (Phasianus colchicus) coming from Poland and Rumania and used in Italy in repopulation interventions for hunting purposes were examined. From each animal the trachea, oesophagus, crop and intestine were collected and examined for the presence of nematodes. The examination of the oesophagi and crops of 5 birds revealed the presence of parasites pertaining to the family Capillariidae: Eucoleus contortus was found in all of the 5 animals, E. annulatus was present along with the previous parasite in one animal coming from Poland. The examination of the tracheae revealed the presence of Syngamus trachea in 5 animals (9.80%). Adult or larval stages of Heterakis gallinarum (37.25% of birds) and Capillariidae (35.29%) were found in the intestinal tracts. Aonchotheca caudinflata was detected only in one bird coming from Poland; the capillarids found in all of the remaining pheasants exhibited morphological characteristics referable to Capillaria phasianina, a species never reported in Italy. The release of game from foreign countries, therefore, may always constitute a risk for the autochthonous one due to the spread of new parasitic infections. PMID- 16252481 TI - Detection of Babesia caballi in Amblyomma variegatum ticks (Acari: Ixodidae) collected from cattle in the Republic of Guinea. AB - A reverse line blot hybridisation (RLB) assay was applied to screen Amblyomma variegatum adult ticks (n = 504) collected from N'Dama cattle in the Republic of Guinea. In a PCR, the V1 hypervariable region of the 16S ribosomal RNA (rRNA) gene was amplified with a set of primers unique for species of the genera Anaplasma and Ehrlichia, and the V4 hypervariable region of the 18S rRNA gene was amplified with primers specific for members of the genera Theileria and Babesia. Amplified PCR products from A. variegatum ticks were hybridised onto a membrane, to which oligonucleotide probes species-specific for Ehrlichia/Anaplasma and Theileria/Babesia parasites were covalently linked. No pathogens belonging to Ehrlichia/Anaplasma species were found, while 10 DNA samples resulted positive for Babesia caballi and 5 samples for Theileria velifera. This is the first report of B. caballi in A. variegatum ticks. One of the B. caballi positive samples was sequenced. This new strain (BcabGuinea) showed a 97% similarity to the Z15104 B. caballi GenBank sequence. PMID- 16252482 TI - Public health risk from avian influenza viruses. AB - Since 1997, avian influenza (AI) virus infections in poultry have taken on new significance, with increasing numbers of cases involving bird-to-human transmission and the resulting production of clinically severe and fatal human infections. Such human infections have been sporadic and are caused by H7N7 and H5N1 high-pathogenicity (HP) and H9N2 low-pathogenicity (LP) AI viruses in Europe and Asia. These infections have raised the level of concern by human health agencies for the potential reassortment of influenza virus genes and generation of the next human pandemic influenza A virus. The presence of endemic infections by H5N1 HPAI viruses in poultry in several Asian countries indicates that these viruses will continue to contaminate the environment and be an exposure risk with human transmission and infection. Furthermore, the reports of mammalian infections with H5N1 AI viruses and, in particular, mammal-to-mammal transmission in humans and tigers are unprecedented. However, the subsequent risk for generating a pandemic human strain is unknown. More international funding from both human and animal health agencies for diagnosis or detection and control of AI in Asia is needed. Additional funding for research is needed to understand why and how these AI viruses infect humans and what pandemic risks they pose. PMID- 16252483 TI - Blackhead disease in turkeys: direct transmission of Histomonas meleagridis from bird to bird in a laboratory model. AB - The spread of Histomonas meleagridis infections through groups of turkeys in the absence of the cecal worm vector (Heterakis gallinarum) was studied in a battery cage model. Battery-reared poults were exposed at 2 wk of age by commingling with infected birds into cages that had the floor lined with paper. One treatment received no exposure, whereas other birds were commingled with two, three, or four birds/cage (25%, 37.5%, or 50%) inoculated per cloaca with cultured H. meleagridis (200,000/bird). Inoculated birds died at 7-13 days postinoculation (DPI) showing typical liver and cecal lesions of histomoniasis. By 14 DPI, 87.5% of the directly inoculated birds died or had severe lesions of histomoniasis. Turkeys commingled with two, three, or four infected birds became infected at the rate of 72%, 80%, or 75%, respectively. In another experiment, two birds/cage (25%) were inoculated with Histomonas from culture and allowed to commingle with other birds for 1, 2, 3, or 4 days. Two of 12 (16.7%) birds had minor cecal lesions after contact with inoculated birds for 1 day, but 87.5%-100% became infected if inoculated birds remained in the cage for 2-4 days. Contemporaneous inoculation with cecal coccidia (Eimeria adenoeides) as a predisposing factor in blackhead infections was studied using the model. Turkey poults directly inoculated with Histomonas were allowed to commingle for 5 days with uninoculated birds that had received inoculation with 0, 10(3), or 10(4) sporulated oocysts. The coccidian infection appeared to interfere with transmission of blackhead infection by 7 DPI, as suggested by lessened severity of cecal lesions and a lower percentage of infected birds. These studies confirm that histomoniasis is transmitted readily from directly exposed young turkeys to others in the absence of the cecal worm vector, and that this phenomenon can be reproduced in battery cages as an experimental model. PMID- 16252484 TI - Viral genotyping of infectious bursal disease viruses isolated from the 2002 acute outbreak in Spain and comparison with previous isolates. AB - An infectious bursal disease (IBD) outbreak occurred in the east region of Spain in the spring of 2002 and rapidly spread thorough the whole country, although proper vaccination programs were applied. In this report, 33 infectious bursal disease viruses (IBDVs) isolated from this outbreak were characterized by nucleotide sequencing of the VP2 gene hypervariable region and were compared with reference IBD strains and the 1990s Spanish IBDVs in order to determine possible emergence of IBDV isolates with modified antigenic or virulent properties. Moreover, histopathologic and immunohistochemical studies of those cases where bursal tissues were available were carried out. Of the 33 isolates, 23 were identified as very virulent IBDVs (vvIBDVs), whereas the other 10 isolates were classified as attenuated or intermediate virulence classical strains and could possibly be IBDV live vaccine strains used in the immunization of these chickens. Results of this study indicate that wIBDV isolates from the 2002 Spanish outbreak are closely related with those from the 1990s outbreak. However, acute IBD cases have not been reported in Spain during these 10 yr. Genetic, management, and environmental factors likely related with IBD reemergence in Spain are discussed. Moreover, our results indicate that good correlation exists between the IBDV subtype present in the field and the degree of lesions in bursa tissue, as well as the immunohistochemistry staining. PMID- 16252485 TI - Development of a polymerase chain reaction-based in vivo method in the diagnosis of subclinical pigeon circovirus infection. AB - This paper describes a polymerase chain reaction (PCR)-based method performed on blood samples and intestinal content to detect subclinical pigeon circovirus (PiCV) infection in live pigeons. In addition, two sets of primers (primer set 1 and 2), designed in two different regions of the viral genome, were used to provide evidence of possible differences in PCR responses. Blood and intestinal content samples were randomly collected from a total of 50 apparently healthy meat pigeons, aged 1 to 5 wk, which came from central Italy. Samples of primary lymphoid organs were also collected. Results showed a high level of PiCV infection, although clinical signs were not present. The results obtained with the two sets of primers showed that primer set 2 was able to detect a higher number of PCR-positive pigeons (45 of 50 pigeons) than primer set 1 (11 of 50 pigeons). In both cases an increase in positive results with pigeon age indicates that the major direction of transmission is likely horizontal. In these circumstances feces can play an important epidemiologic role, as supported by the consistent circovirus detection in intestinal content. The high sensitivity of this PCR test, which is able to detect very low amounts of viral DNA (5.5 x 10( 3) fg of plasmid containing the cloned PiCV genome), makes it suitable for possible application as an epidemiologic tool for identifying virus carriers for subsequent removal from lofts. PMID- 16252486 TI - Partial characterization of an adenovirus-like virus isolated from broiler chickens with transmissible viral proventriculitis. AB - Transmissible viral proventriculitis (TVP) was experimentally reproduced in specific-pathogen-free chickens using a homogenate of proventricular tissue obtained from TVP-affected commercial broiler chickens. Thin-section electron microscopy revealed intranuclear, approximately 70-nanometer (nm), adenovirus like viruses (AdLV) within proventricular lesions. The AdLV, designated AdLV (R11/3), could not be propagated using various avian and mammalian cell cultures or by inoculation of embryonated chicken eggs by yolk, allantoic, or chorioallantoic membrane routes. However, AdLV (R11/3) was successfully propagated by amniotic inoculation of embryonated chicken eggs, with detection of the virus in proventriculi and intestinal contents of hatched 2-day-old chicks (8 days postinoculation). Virus propagation was evident in in ovo-inoculated chicks by (1) gross and microscopic lesions in proventriculi consistent with TVP, (2) immunohistochemical localization of AdLV (R11/3) antigens in proventricular epithelium, (3) thin-section electron microscopic detection of intranuclear, approximately 70-nm AdLVs within proventricular epithelium, and (4) negative stain electron microscopic detection of extracellular, approximately 70-nm AdLVs in intestinal contents. Indirect immunofluorescence and polymerase chain reaction procedures that specifically recognize groups I, II, and III avian adenoviruses failed to recognize AdLV (R11/3). The findings suggest an etiologic role for AdLV (R11/3) in TVP and indicate that this virus is distinct from known avian adenoviruses. PMID- 16252487 TI - Reproduction of proventriculitis in commercial and specific-pathogen-free broiler chickens. AB - Proventriculitis was studied by experimentally reproducing the disease in broiler chickens. One-day-old infectious bursal disease virus (IBDV) and infectious bronchitis virus (IBV) antibody positive commercial broilers and 1-day-old antibody negative specific-pathogen-free (SPF) broilers were orally gavaged with proventricular homogenates produced from the proventriculi of broilers with proventriculitis. At 7 and 14 days, both commercial and SPF broilers had enlargement of the proventriculus with necrosis of the glandular epithelium and lymphocytic infiltrates in the proventricular glands. SPF broilers exposed to the proventricular homogenates developed infectious bursal disease, and IBDV was detected by reverse transcriptase-polymerase chain reaction (RT-PCR). They also were positive by RT-PCR to IBV and developed nephritis. Commercial broilers developed mild nephritis but not bursal disease and were negative for IBDV and positive for IBV by RT-PCR. Both homogenate-inoculated commercial and SPF chickens were negative for reovirus and Newcastle disease virus by RT-PCR and variably positive for adenovirus by PCR. Bacteria were not identified in histologic sections, nor were they isolated from affected proventriculi. Indirect fluorescent antibody assay using convalescent sera detected intracytoplasmic staining in the proventricular glandular epithelial cells. Examination of thin sections of proventriculi using electron microscopy identified virus-like particles approximately 120 nm in diameter within the cytoplasm of these cells at 7 days after inoculation. Passage of proventricular homogenate filtrates in chicken embryos for virus isolation caused stunting, and allantoic fluid from these eggs was positive for IBV by RT-PCR. PMID- 16252488 TI - Effect of growth promotant usage on enterococci species on a poultry farm. AB - The species population of enterococci isolated from four poultry houses for six grow-outs on one farm was determined. Two houses on the farm were control houses and did not use any antimicrobials, while two other houses on the farm used flavomycin, virginiamycin, and bacitracin during different poultry grow-outs. Litter, chick boxliners, feed, and poultry carcass rinses were obtained from each house and cultured for the presence of enterococci. Nine species of enterococci (Enterococcusfaecalis, E. faecium, E. avium, E. casselifiavus, E. cecorum, E. durans, E. gallinarum, E. hirae, and E. malodoratus) were identified from the study. Enterococcus faecalis was isolated more frequently from chick boxliners (n=176; 92%) and carcass rinses (n=491; 69%), whereas E. faecium was found more frequently in litter (n=361; 77%) and feed (n=67; 64%). Enterococcus faecalis (n=763; 52%) and E. faecium (n=578; 39%) were isolated most often from the farm and houses, regardless of antimicrobial treatment. Fifty-two percent of E. faecalis and 39% of E. faecium were isolated from both control (n=389 and 295, respectively) and treatment (n=374 and 283, respectively) samples. This study indicates that antimicrobial usage on this farm did not alter the resident population of enterococci. PMID- 16252489 TI - Comparison of the specificity and sensitivity of PCR, nested PCR, and real-time PCR for the diagnosis of histomoniasis. AB - Blackhead, also known as enterohepatitis, is caused by a protozoan parasite called Histomonas meleagridis. Clinical symptoms are nonspecific. Until now, diagnosis has been mainly based on postmortem lesions and microscopical and histopathological examination. In many cases, especially in layer flocks, these conventional methods are not sufficient, as the lesions are sometimes not clear. The technique for isolation of histomonads in vitro offers many advantages, but the confirmation of histomonads growing in culture may require a time-consuming procedure of rectal inoculation of culture material into chickens or turkeys. The aim of our investigation was to establish a conventional polymerase chain reaction (PCR), a nested PCR, and a real-time PCR, and to examine their specificity as well as sensitivity in the diagnosis of histomoniasis. The obtained results have shown that the conventional PCR is more sensitive than the real-time PCR. Furthermore, the sensitivity of the PCR can be increased by adding the nested PCR. However, the real-time PCR is more specific. PMID- 16252490 TI - Serologic evidence of West Nile virus infection in three wild raptor populations. AB - We assayed for West Nile virus (WNV) antibodies to determine the presence and prevalence of WNV infection in three raptor populations in southeast Wisconsin during 2003-04. This study was conducted in the framework of ongoing population studies that started before WNV was introduced to the study area. For 354 samples, 88% of 42 adult Cooper's hawks (Accipiter cooperii), 2.1% of 96 nestling Cooper's hawks, 9.2% of 141 nestling red-tailed hawks (Buteo jamaicensis), and 12% of 73 nestling great horned owls (Bubo virginianus) tested positive for WNV antibodies by the constant virus-serum dilution neutralization test. Samples that tested positive for WNV antibodies were collected across a wide variety of habitat types, including urban habitats (both high and low density), roads, parking areas, recreational areas, croplands, pastures, grasslands, woodlands, and wetlands. Based on the increased prevalence and significantly higher WNV antibody titers in adults compared with nestlings, we suggest that nestlings with detectable antibody levels acquired these antibodies through passive transmission from the mother during egg production. Low levels of WNV antibodies in nestlings could serve as a surrogate marker of exposure in adult raptor populations. Based on breeding population densities and reproductive success over the past 15 yr, we found no apparent adverse effects of WNV infections on these wild raptor populations. PMID- 16252491 TI - Characterization of the neurotoxin produced by isolates associated with avian botulism. AB - Several varieties of birds are affected by type C botulism. We conducted neutralization tests of culture supernatants of isolates from cases of avian botulism. Whereas the toxin produced by isolates derived from mammalian botulism was neutralized only with type C antitoxin, the toxins of all isolates related to avian botulism were neutralized with both type C and D antitoxins. An analysis of nucleotide sequences with several strains revealed that the neurotoxin gene in the isolates from avian botulism comprises two thirds of the type C neurotoxin gene and one third of the type D neurotoxin gene. This indicates that the neurotoxin of avian isolates is a mosaic of type C and D neurotoxins. We prepared three sets of primers to differentiate the gene for the mosaic form from the conserved genes of type C and D neurotoxins. The results of polymerase chain reaction with these primers indicated that all avian botulism-related isolates and specimens possess the gene for the mosaic form of the neurotoxin. The toxins purified from avian and mammalian isolates exhibited the same degree of lethality in mice, but the former showed greater toxicity to chickens than the latter. These results indicate that the mosaic neurotoxin is probably a pathogenic agent causing some forms of avian botulism. PMID- 16252492 TI - The relationship between the duration of fecal shedding and the production of contaminated eggs by laying hens infected with strains of Salmonella enteritidis and Salmonella Heidelberg. AB - Egg contamination by Salmonella Enteritidis has remained a significant public health problem for nearly two decades, and Salmonella Heidelberg has also been recently implicated in egg-transmitted human illness. Colonization of the intestinal tract is a necessary precursor to the invasion of reproductive organs and subsequent deposition inside eggs laid by infected hens, but the relationship between the persistence of Salmonella in the intestinal tract and the likelihood of egg contamination has been uncertain. In this study, groups of laying hens were inoculated with large oral doses of strains of Salmonella Enteritidis and Salmonella Heidelberg, including variants of the original parent strains that had been reisolated from eggs laid by infected hens in a prior study. The shedding of Salmonella in voided feces was monitored for 6 wk postinoculation, and all eggs laid by infected hens between 5 and 22 days postinoculation were cultured for Salmonella in their contents. The mean duration of fecal shedding was significantly longer for the previously passaged Salmonella strains (26.7 days) than for the original parent strains (17.5 days), and the passaged strains caused a significantly higher frequency of egg contamination (6.4%) than did the parent strains (3.3%). However, the duration of fecal shedding and the frequency of egg contamination were not correlated for any of the Salmonella Enteritidis or Salmonella Heidelberg strains. PMID- 16252493 TI - Monitoring presence and annual variation of trichomoniasis in mourning doves. AB - Information about the annual variation of trichomoniasis in mourning doves (Zenaida macroura) may be important in understanding mechanisms affecting mourning dove populations. The objectives of this study were to monitor the presence and annual variation of Trichomonas gallinae for 6 yr in a local mourning dove population using hunter-killed doves. During 1998-2003, 4052 hunter killed doves were sampled for the presence of T. gallinae; 226 (5.6%) tested positive (4.4%-10.6% range). Results of the monitoring effort were relatively consistent during the 6-yr period, with the presence of T. gallinae being within the range of previously reported estimates for mourning doves. Asymptomatic carriers in one segment of the dove population may provide a mechanism for spreading the disease to other segments of the mourning dove population. PMID- 16252494 TI - Dynamics of Campylobacter spp. spread investigated in 14 broiler flocks in Switzerland. AB - Ten conventional and four extensive outdoor broiler flocks, distributed over nine farms, were investigated twice per week during a 35-58-day rearing period to observe the dynamics of Campylobacter spp. spread within these flocks. Strains isolated during this period were genotyped by restriction fragment length polymorphism analysis of the flaA gene and macrorestriction profiling with pulsed field gel electrophoresis. A total of 4112 samples were collected; 157 (3.8%) of these samples were Campylobacter positive, with all C. jejuni. The positive samples were distributed over three conventional and two extensive outdoor flocks on five farms. These five positive flocks were colonized from the fifth to the seventh week of age and remained colonized until slaughter. Each of the flocks showed a flock-specific genotype of Campylobacter that predominated until slaughter. Presuming different ways of entry, a combination of this fact and the observed dynamics of C. jejuni spread within the flocks indicates that a single source from the environment may have been responsible for the colonization of each flock. These conclusions may serve to further develop combat strategies at farm level. PMID- 16252495 TI - Development of quantitative real-time polymerase chain reaction for duck enteritis virus DNA. AB - Duck enteritis virus (DEV) is a herpesvirus that causes an acute, contagious, and fatal disease. In the present article, we introduce a quantitative real-time polymerase chain reaction (PCR) assay for DEV DNA using TaqMan technology and a two-step protocol. It was confirmed to be rapid, sensitive, and specific for DEV detection. The primers and probe were designed and directed to the DNA polymerase gene of DEV. The method will provide a valuable tool for rapid laboratory diagnosis of DEV infection. By virtue of its high-throughput format and its ability to accurately quantify the viral DNA, the method may be useful for large epidemiological surveys and clarification of pathogenesis, such as latency and reactivation of the virus. PMID- 16252496 TI - Construction and characterization of a fowlpox virus field isolate whose genome lacks reticuloendotheliosis provirus nucleotide sequences. AB - Fowlpox virus (FWPV) has been isolated from vaccinated chicken flocks during subsequent fowlpox outbreaks that were characterized by a high degree of mortality and significant economic losses. This inability of current vaccines to induce adequate immunity in poultry could be reflective of an antigenic and/or biologic distinctiveness of FWPV field isolates. In this regard, whereas an infectious reticuloendotheliosis virus (REV) provirus is present in the majority of the field viruses' genomes, only remnants of REV long terminal repeats (LTR) have been retained in the DNAs of each vaccine strain. Although it has not been demonstrated whether the partial LTRs can provide an avenue for FWPV to reacquire the REV provirus by homologous recombination, utilizing viruses of which genomes lack any known integrated retroviral sequences could resolve concern over this issue. Therefore, such an entity was created by genetically modifying a recently isolated field strain of FWPV. This selection, in lieu of a commercial vaccine virus, as the progenitor was based on the probability that a virus circulating in the environment would be more antigenically similar to others in this locale and thus might be a better candidate for vaccine development. A comparison in vivo of the pathogenic traits of the parental wild-type field isolate, its genetically modified progeny, and a rescue mutant in whose genome the REV provirus was inserted at its previous location, indicated that elimination of the provirus sequence correlated with reduced virulence. However, even with elimination of the parasitic REV, the modified FWPV was still slightly more invasive than a commercial vaccine virus. Interestingly, both types of attenuated FWPV elicited a similar degree of antibody production in inoculated chickens and afforded them protection against a subsequent challenge by a field virus, the origin of which was temporally and geographically distinct from that of the progenitor strain. Due to its antigenicity being retained despite a decrease in virulence, this REV less FWPV could potentially be developed as a vaccine against fowlpox. PMID- 16252497 TI - Phenotypic characterization of ipaH+ Escherichia coli strains associated with yolk sac infection. AB - Seventy-six Escherichia coli serotypes possessing the ipaH gene typical of enteroinvasive E. coli (EIEC) strains were characterized. Biochemical identification of our strains shows positive reactions for lactose fermentation (100% of strains), lysine decarboxylase (98.7% of strains) and motility (67.1% of strains), properties that do not correspond with those described to the EIEC group. The serotypes agree with an initial classification. In this, some common O antigens identified among ipaH+ strains were O2 (n=20), OR (n=11) and non determined O? (n=10). The O2:NM serotype was the most common. Sixty-six percent (n=50) of the ipaH+ E. coli strains were colicin producers, of them, 26 (34%) produced Col V and other colicins, 13 (17%) produced colicins other than Col V, and 11 (14.5%) produced Col V only. Trimethoprim/Sulfa (72%), ampicillin (64.5%), enrofloxacin (55.3%), and ciprofloxacin (47.4%) were the major antimicrobial resistance frequencies observed. Twenty-five different multiresistance patterns were observed, where sixty-six strains (86.8%) were included. A MIC test showed that most of the strains were sensitive to low gentamicin and kanamycin concentrations, whereas most of the strains were resistant to tetracycline. An invasiveness assay showed that the predominant alterations caused to HEp-2 cells were changes in shape and staining, and in most of the specimens, a partial monolayer detachment was also seen. Fifteen strains invaded more than 30% of the monolayer cells, causing the formation of intercellular bridges or filipoidal like protrusions. The results suggest the existence of specific clone complexes derived from EIEC strains adapted to the avian host. To our knowledge, this is the first study that demonstrates the presence of extraintestinal invasive E. coli (ExIEC) strains. PMID- 16252498 TI - Comparison of a Salmonella enteritidis-specific polymerase chain reaction assay to delayed secondary enrichment culture for the detection of Salmonella enteritidis in environmental drag swab samples. AB - The California Egg Quality Assurance Program uses the delayed secondary enrichment culture method for detecting Salmonella Enteritidis in environmental drag swabs obtained from commercial layer complexes. The turnaround time for this method is variable and is dependent on the prevalence of Salmonella, level of the Salmonella identification, and capabilities of the performing laboratory. On a sample basis, a range of 4 to 8 days is required to identify a Salmonella sp. to the serogroup level. Additional time is required to serotype group D Salmonella isolates. A Salmonella Enteritidis-specific polymerase chain reaction (PCR) assay was developed for use on drag swabs and chick box papers, and had a turnaround time of 3-4 days. The delayed secondary enrichment culture method and the Salmonella Enteritidis-specific PCR assay were compared on 942 drag swab and 85 chick box paper samples submitted from 217 and 22 premises, respectively, as part of the California Egg Quality Assurance Program. The PCR assay identified 43 positive Salmonella Enteritidis samples from 22 premises, whereas the culture method identified 24 group D Salmonella-positive samples from 16 premises. There was a significant difference (P = 0.001) in the proportion of positive samples as determined by the two assays. Complete serotyping of the group D Salmonella positive cultures confirmed Salmonella Enteritidis in all but one sample that was identified as Salmonella Jamaica and was negative by the PCR assay. PMID- 16252499 TI - Evaluation of dietary Natustat for control of Histomonas meleagridis in male turkeys on infected litter. AB - Histomoniasis (histomonosis, infectious enterohepatitis, or blackhead) is a disease of turkeys on litter or range caused by the protozoan Histomonas meleagridis, a parasite of worms, primarily spread in feces, in Heterakis gallinarum (cecal worm) eggs, or in Eisenia foetida (earthworms). In this trial, Natustat (Alltech, Inc., Nicholasville, KY), a proprietary plant-derived product, was used at 1.925 kg/tonne and compared with nitarsone in male hybrid turkey diets to 42 days of age on histomonad infected litter (day 7) from broiler breeders. Infected nonsupplemented and uninfected nonsupplemented control groups were also included. Natustat and nitarsone significantly improved 28- and 42-day feed conversion ratios and lowered 28- and 35-day cecal and liver lesion scores compared with infected nonsupplemented turkeys. The body weight at 42 days was greater in the Natustat and nitarsone supplemented groups than in the infected nonsupplemented group. PMID- 16252500 TI - Acquisition of immunity to Eimeria maxima in newly hatched chickens given 100 oocysts. AB - The acquisition of immunity to Eimeria maxima by chicks infected 18 hr after hatch with a single dose of 100 oocysts was investigated. In the first experiment, birds were moved each day to clean cages in order to prevent the possibility of secondary infection resulting from ingestion of oocysts passed in their feces. Immunity was measured at 4 wk of age by calculation of oocyst production following challenge with 500 oocysts or weight gain following challenge with 100,000 oocysts. Large numbers of oocysts were produced by infected birds following challenge, although numbers were significantly less than those from birds that had been reared in the absence of infection (susceptible controls). The weight gain of infected birds following challenge was significantly greater than that of susceptible controls but less than that of unchallenged controls. Thus, only partial protection had been acquired, whether parasite replication or body weight gain was used to assess the extent of immunity development. In a second experiment, acquisition of immunity at 4 wk by chicks infected 18 hr after hatch with 100 oocysts of E. maxima and reared in floor pens in contact with their droppings was investigated. Infected birds produced no oocysts following challenge, and weight gains were not significantly different from the unchallenged controls, which indicates that full immunity had developed by 4 wk. It is concluded that if oocysts of Eimeria species are used to vaccinate day-old chicks, reinfection by oocysts present in the litter is necessary for the establishment of protective immunity. PMID- 16252501 TI - Metastatic adenocarcinoma in the lungs of older laying hens. AB - As part of a study on ovarian and oviductal adenocarcinomas in older laying hens, 676 commercial egg-laying chickens were necropsied at 4 yr of age. Tumors were identified in 305 (45.1%) of the hens. Grossly visible metastatic tumors in the lungs of a few birds prompted collection of lungs from 228 affected hens for histologic examination. Metastatic adenocarcinomas were identified histologically in the lungs of 42 of the 228 hens (18.4%). The number of tumors and extent of involvement of the lung parenchyma varied from minimal to extensive. Tumor emboli were occasionally seen in pulmonary vessels. Metastatic foci showing expansive or invasive growth were identified. These findings indicate that vascular spread of adenocarcinomas is more common than previously thought in chickens, and the lungs are often involved with metastatic tumors, primarily from the reproductive tract. PMID- 16252502 TI - Malaria in an eastern screech owl (Otus asio). AB - Owls are frequent carriers of blood parasites but clinical malaria infections are rare. Various stages of Plasmodium subpraecox were seen in 90% of the erythrocytes of an Eastern screech owl (Otus asio) showing symptoms consistent with malaria 1 wk after admission for traumatic injuries. An additional unidentified blood parasite, either a Plasmodium or a Haemoproteus spp. was found in small numbers of red blood cells on blood films examined at admission and at day 7 postadmission. Combined infestation, trauma-induced stress, and iatrogenic corticosteroid administration are possible factors that could have induced disease. Oral treatment with mefloquine at 30 mg/kg, repeated after 12, 24, and 48 hr, proved successful in eliminating both organisms and signs of clinical disease. PMID- 16252503 TI - Epidemiology, pathology, and immunohistochemistry of layer hens naturally affected with H5N1 highly pathogenic avian influenza in Japan. AB - Epidemiology, pathology, and immunohistochemistry were investigated in layer hens affected with H5N1 highly pathogenic avian influenza, which occurred for the first time in 79 years in Japan. The farm, which had a total of 34,640 chickens, experienced up to 43.3% mortality before the chickens were depopulated. Clinically, the affected chickens exhibited mortality without apparent clinical signs. Histologically, hepatocytic necrosis; necrosis of ellipsoids and follicles with fibrin in the spleen; necrosis with glial nodules in the brain stem, cerebrum, and cerebellum; necrosis of acinar cells in the pancreas; and necrosis of lymphoid tissues in intestinal lamina propria were seen. Occasionally, mild bronchiolitis, degeneration of smooth muscle fibers in the cecum, and mild tubulonephrosis were noted. Immunohistochemically, influenza virus antigens were detected often in the liver and spleen, heart, intestine, gizzard, proventriculus, and oviduct. In addition, antigens were seen also in the brain, kidney, pancreas, and ovary, but seldom in the lung and trachea. Virus antigen was mainly detected in the capillary endothelium and parenchymal cells. This suggests that virus excretion from the respiratory tract was not as prevalent as that from the digestive tract in the present cases. PMID- 16252504 TI - Outbreak of avian mycobacteriosis in a flock of two-year-old domestic pigeons (Columba livia f. domestica). AB - This report describes an outbreak of avian mycobacteriosis in a flock of 100 two yr-old pigeons. Over a 6-mo period, the sick pigeons showed cachexia followed by death. In Columbiformes classic tubercles rarely develop, but in these affected pigeons granulomatous nodular lesions of various sizes, containing numerous acid fast bacilli, were found in the internal organs. The lesions were observed in the liver, spleen, intestine, bone marrow, ovary, and oviduct. Despite their breeding age, atrophy was also found in the ovary and oviduct. Microorganisms belonging to Mycobacterium avium complex were identified in the affected tissues by polymerase chain reaction. PMID- 16252506 TI - Summary of influenza activity, September 2004 - August 2005. PMID- 16252505 TI - Isolation and preliminary characterization of chicken anemia virus from chickens in Nigeria. AB - Chicken anemia virus (CAV) was isolated for the first time from the Nigerian chicken population. The virus was recovered from necropsied birds from broiler and pullet flocks that suffered disease outbreaks tentatively diagnosed as infectious bursal disease. A sensitive polymerase chain reaction (PCR) assay detected CAV DNA in tissues of necropsied birds. Restriction endonuclease analysis performed with the 733-bp PCR product and the Cfo I enzyme indicated at least two different CAVs were circulating among the Nigerian chicken population. Four isolates were obtained from pooled liver and thymus tissues using the MDCC MSB1 cell line. These isolates were found to be antigenically closely related to the Cuxhaven-1 (Cux-1) reference strain of CAV when reacted with four monoclonal antibodies prepared against the Cux-1 virus. One of the isolates (isolate A) induced thymus atrophy, bone marrow aplasia, and low hematocrit values when inoculated into 1-day-old specific-pathogen-free chickens. These findings not only demonstrate that CAV is present in Nigeria, but they also likely represent the first cell culture isolation of the virus in Africa. PMID- 16252507 TI - Eleventh informal consultation of the global polio laboratory network, 30 August 1 September 2005. PMID- 16252508 TI - [Prescription by INN (International Non-Proprietary Name)]. PMID- 16252509 TI - [The endocannabinoid system and the regulation of the metabolism]. AB - The endocannabinoid system modulates many physiological functions by acting on receptors CB1 and CB2. The endocannabinoids are produced only when and where they are needed. They act locally and are immediately metabolised after their action. Overactivation of the endocannabinoid system is observed in obesity, with stimulation of the appetite in the hypothalamus and fat accumulation in the adipocytes with increase of insulin resistance and decrease of adiponectin. Nicotine use overactivates also the endocannabinoid system. CB1 blockade by a specific inhibitor (rimonabant) decreases food intake and weight in animal studies and increases adiponectin and insulin sensitivity. Moreover, rimonabant decreases tobacco dependence. Clinical studies with rimonabant are encouraging. PMID- 16252510 TI - [Rimonabant (Acomplia), specific inhibitor of the endocannabinoid system]. AB - The endocannabinoid system plays a major role in the regulation of body energy by stimulation of the appetite in the hypothalamus and increase of fat accumulation in adipocytes. The blockade of the cannabinoid system (CB1) by the specific inhibitor (rimonabant) decreases food intake and adiposity in animals and in humans. Moreover rimonabant lowers tobacco addiction. Clinical studies (RIO LIPIDS and RIO-EUROPE) have recently confirmed that rimonabant combined with a hypocaloric diet over 1 year, promoted significant decrease of body weight, waist circumference and improvement of dyslipidemia. Rimonabant was well tolerated with mild and transient side effects. The future place of rimonabant in the strategy of obesity is still to be clarified. PMID- 16252511 TI - Piracetam preparations on the Belgian market: a comparative study. AB - Piracetam preparations formulated as capsules, tablets and granules were evaluated with different tests including in vitro dissolution and assay with previously validated methods according to the guideline of the European network of official medicines control laboratories (OMCLs). All examined products complied with the requirements as described here and in the European Pharmacopoeia except for the dissolution test where 3 products did not meet the USP acceptance criteria applied on the established specification. PMID- 16252512 TI - [Ophthalmic preparations from ancient times until today]. PMID- 16252513 TI - A list of the mosquitoes housed in the mosquito museum at the Vector Control Research Centre, Pondicherry, India. AB - The establishment of a mosquito museum at the Vector Control Research Centre, Pondicherry, India, is reported. The collection at present contains more than 14,800 adult mosquitoes, including 8,426 that are individually pinned. The collection also includes some 1,048 male and female genitalia preparations, and 815 larval and 444 pupal exuviae on microscope slides. Mosquitoes of 19 genera, 37 subgenera, and 181 species are housed in the museum, representing about 50% of the known species from India. A list of the species is provided. PMID- 16252514 TI - List of species described in the egg stage of tribe Aedini (Diptera: Culicidae) with their literature citations. AB - Species of Aedini with published morphological descriptions and/or illustrations of eggs are listed with their literature citations. Each species has a notation indicating descriptions are written, illustrated or both. Only approximately 16% of aedine species currently have morphological descriptions published for the egg stage. The new classification of tribe Aedini proposed by Reinert et al. (2004) is utilized. PMID- 16252515 TI - Attraction, feeding, and repellency responses in mutant strains of Aedes aegypti. AB - In a laboratory olfactometer, 12% of female Aedes aegypti with a marker gene for red eye (re), 0.7% of females with a marker gene for white eye (we), and 54.1% of females with normal (norm) eye color were attracted to odor from a human hand. When a synthetic attractant blend was used in place of the hand, the attraction rate was 7%, 0.3%, and 35.4%, respectively. On average, re females required significantly less time (76.8 sec) than we (189.6 sec) or norm (176.7 sec) females to locate, land on, and probe human skin but no difference was found between mosquito strains in the time required for females to bloodfeed to repletion on a restrained guinea pig. Differences among mosquito strains in the repellency of 15% diethyltoluamide (deet), 6.65% deet, and 10% citronella were not significant. PMID- 16252516 TI - Evaluation of oviposition substrates and organic infusions on collection of Culex in Florida. AB - Gravid mosquito traps are commonly used for both arbovirus surveillance and population surveillance of mosquitoes of the genus Culex. Oviposition substrates, used as baits in these traps, were tested against Culex under laboratory and field conditions. In the laboratory all substrates tested as 1% and 10% dilutions in 2-choice bioassays against female Cx. quinquefasciatus were significantly more effective than well water controls in eliciting oviposition. Strongest responses were to dilutions of dairy effluent, followed by larval water and infusions of alfalfa hay, alfalfa pellets, Bermuda hay, oak leaves, and Typha leaves, with lowest responses to cow manure infusion. In the field, few significant differences in collections were obtained between traps baited with different infusions. Significantly more Cx. quinquefasciatus and Cx. nigripalpus were collected in traps baited with cow manure infusion (highest) compared to alfalfa hay infusion (lowest). Responses of Cx. quinquefasciatus to dairy effluent and infusions of Bermuda hay, oak leaves, and Typha leaves were not significantly different from either cow manure infusion or alfalfa hay infusion. Responses of Cx. nigripalpus were highest to cow manure infusion and equally low to infusions of alfalfa hay and Typha leaves; moderate responses were observed to dairy effluent and infusions of Bermuda hay and oak leaves. Gravid females comprised 66.7-81.9% of the collections for each infusion type, with no significant difference among infusions in the proportion of gravid females collected. PMID- 16252517 TI - Relative abundance of tree hole-breeding mosquitoes in Boone County, Missouri, USA, with emphasis on the vector potential of Aedes triseriatus for canine heartworm, Dirofilaria immitis (Spirurida: Filariidae). AB - Aedes (Protomacleaya) triseriatus currently shares its habitat in the USA with the introduced species Aedes (Finlaya) japonicus and Aedes (Stegomyia) albopictus. In the late 1980s, before the introduction of these 2 species, Ae. triseriatus was the dominant tree hole- and artificial container-breeding mosquito in central Missouri. Aedes triseriatus represented 89% of the mosquito immatures collected from water-filled tree holes and artificial containers at 3 forested field sites in central Missouri, from May to October, 1986 to 1988. Laboratory-reared female Ae. triseriatus were able to support larval development of Dirofilaria immitis (canine heartworm) to the infective 3rd larval stage. A blood meal from a microfilaremic Collie-mix dog was sufficient to infect adult female mosquitoes, indicating that Ae. triseriatus is a possible vector of canine heartworm in central Missouri. Confirmation of the vector status of this species depends on the yet-to-be observed transmission of D. immitis by Ae. triseriatus in the field, possibly by experimental infection of dogs by wild-caught mosquitoes. Defining the role of this species in epizootic outbreaks could contribute toward accurate risk assessment as the abundance of Ae. triseriatus increases and decreases in response to the success of Ae. albopictus, Ae. japonicus, or other introduced container-breeding mosquitoes. PMID- 16252518 TI - Comparative susceptibility of three species of Anopheles from Belize, Central America, to Plasmodium falciparum (NF-54). AB - In August of 2000, a comparative susceptibility study was conducted using 3 species of Anopheles mosquitoes from Belize, Central America, and a standard species used in laboratory infection studies, Anopheles stephensi. Test populations were fed human blood infected with cultured Plasmodium falciparum (NF 54 strain) parasites via a membrane feeder. The control species, An. stephensi, exhibited the highest infections, with 73.8% of dissected specimens positive for sporozoites in the salivary glands. The control species also showed heavier sporozoite loads; 74.0% of positive glands having greater than 200 sporozoites. Of species from Belize, Anopheles darlingi was the most susceptibile, e.g., 41.0% of salivary glands were positive, with more than 200 sporozoites per gland. Anopheles vestitipennis had a low salivary gland infection rate (9.3%) and a moderate number of sporozoites in glands (i.e., 85.7% containing 50-250 sporozoites). Anopheles albimanus was the least susceptible species to infection. No specimens of An. albimanus from the Golden Stream population developed sporozoites in the salivary glands, yet 20.7% of dissected specimens had positive midgut infections. The An. albimanus Buena Vista population showed similar results with only a 2.2% salivary gland infection rate and a 21.5% midgut infection rate. Oocysts in An. stephensi increased in size by 20% after day 10. Development peaked at day 12, with a mean oocyst diameter of 58 microm at onset of oocyst differentiation. Oocysts developed more slowly in An. vestitipennis until day 10. After day 10, there was a 53% increase in oocyst development over the previous 10 days. Oocyst differentiation was not observed until day 13 postfeed. As with An. vestitipennis, both populations of An. albimanus showed similar slow rates of oocyst development; however, no dramatic growth increase occurred after day 10. The oocysts in the Golden Stream population exhibited a cessation of growth after day 10, peaking at a mean of 30 microm. The Buena Vista population did not exhibit the same level of reduced oocyst development. A gradual increase in growth continued until days 13 and 14 (36.7 and 35.7 microm, respectively). PMID- 16252519 TI - Low pilot exposure to pyrethrin during ultra-low-volume (ULV) aerial insecticide application for control of adult mosquitoes. AB - Pilot exposure to pyrethrin was monitored after 2 aerial applications of a ultra low-volume (ULV) pyrethrin insecticide for the control of adult mosquitoes. Pyrethrin exposure was estimated by measuring the excretion in urine of a common metabolite, trans-chrysanthemumdicarboxylic acid, of the natural pyrethrin mixture. Pyrethrin exposure estimated by total daily urine volume was well correlated (R2 = 0.8) with exposure estimated by the creatinine-adjusted volume of combined workday urine voids, indicating that a postapplication spot urine sample would be sufficient to measure pyrethrin exposure. Pilot exposure to pyrethrin was very low after both insecticide applications. The highest exposure was found on day 1, with a dose of 2.05 microg pyrethrin equivalents/day or a dosage of 0.03 microg pyrethrin equivalents/kg/day. These exposure rates represent approximately 1/2,800,000th of the low observed adverse effect level and 1/1,000th of the acceptable daily intake for pyrethrin. The aerial application of ULV pyrethrin insecticide for the control of adult mosquitoes does not result in undue exposure to a pilot who is trained and certified to conduct such control operations. PMID- 16252521 TI - Aerial ultra-low-volume assessment of ANVIL 10+10 against Anopheles quadrimaculatus. AB - Against caged field-collected adults of Anopheles quadrimaculatus, optimized aerial applications of ANVIL 10+10 at 18.4 ml (1.63 g active ingredient)/0.4 ha produced a mean mortality of 92% at 1, 12, and 24 h after treatment. These trials were conducted with 7 stacked passes applied 1,067 m upwind of the target site when wind velocity was 18-21 km/h at an application altitude of 46 m and 5-10 km/h at groundlevel. When the wind velocity aloft was greater than might be operationally acceptable (27 km/h), reduced efficacy (64% mortality) was observed in the bioassay site (1,524 m downwind from 11 stacked passes) in an unreplicated application. Droplets from the 2 tail-mounted Beecomist 360-A-12-V rotary atomizers (fitted with wire sleeve cages of size 40 mesh) collected on spinning Teflon-coated slides in the target area averaged a volume mean diameter (VMD) of 29 microm and a droplet density of 39 droplets/cm2 in 18- to 21-km/h winds. With 27-km/h winds aloft and a greater application offset, the VMD and droplet density dropped to 23 microm and 18 droplets/cm2, respectively. Coupled with unreplicated results from a nonoptimized application, these tests provide further documentation of the efficacy of ANVIL 10+10 for wide-area adult mosquito control and for optimization technology. PMID- 16252520 TI - Laboratory and field evaluation of the insect growth regulator pyriproxyfen (Sumilarv 0.5G) against dengue vectors. AB - The insect growth regulator pyriproxyfen was tested against Aedes aegypti at 0.01 and 0.02 mg of active ingredient (AI) per liter of water in 60-liter earthern jars. Both concentrations provided 100% control for 4 months. In additional experiments where 10 liters of water were replaced fortnightly, 100% control was still obtained over 4 months with 0.02 mg AI/liter and greater than 93-100% control was obtained over 4 months with 0.01 mg AI/liter. In less-controlled field-trial conditions, pyriproxyfen at a dosage of 0.02 mg AI/liter provided 100% control for 10 wk against Aedes albopictus even though water was replaced either daily or weekly. Although the activity of pyriproxyfen declines after 10 wk, those tests in the plastic tubs showed much higher levels of sustained residual activity compared to those in the earthern jars. Pyriproxyfen did not have an impact on nontarget organisms. PMID- 16252522 TI - Insecticide resistance in Culex pipiens from New York. AB - Insecticides are the primary means to control Culex pipiens, an enzootic vector of West Nile virus, in the USA. To better understand how the evolution of resistance might impact control of this insect, we investigated the levels of resistance in populations collected from 2 metropolitan areas (Albany and Syracuse, NY) to 4 larvicides (methoprene, phenothrin, Bacillus sphaericus [Bs], and Bacillus thuringiensis israelensis [Bti]) and 1 adulticide (phenothrin) registered for mosquito control in New York State. High levels of resistance were found only to Bti, and only at 1 site (Syracuse). Resistance levels to the other insecticides were less than 10-fold. Given the large difference in Bti resistance between Syracuse and Albany, it appears these populations of Cx. pipiens do not rapidly mix, leading to localization of resistant populations. PMID- 16252523 TI - Development of Triatoma rubida sonoriana, Triatoma barberi, and Meccus mazzottii (Heteroptera, Reduviidae) under laboratory conditions. AB - The objective of this study was to determine some of the most important bionomic parameters related to the capacity of transmission of Trypanosoma cruzi and to estimate the potential transmission capacity and role of 3 groups of recent colonized Mexican Triatoma barberi, T. rubida sonoriana, and Meccus (formerly Triatoma) mazzottii in the prevalence of Chagas disease cases based on the distribution area of each species. Among the studied Triatominae species, the life cycle was shorter in T. r. sonoriana and longer in T. barberi, by as much as 4 times. The 1st and 5th instars had the highest rates of mortality in most of the studied species. Statistically significant shorter duration of feeding of instars was observed for T. r. sonoriana and longer duration was found for M. mazzottii when the species were compared. The mean number of blood meals per nymphal stadium was statistically greater in T. barberi than in T. r. sonoriana and M. mazzottii. Triatoma r. sonoriana and M. mazzotti had shorter defecation delay than in T. barberi and the delay was uniform in all instars. Most of the studied parameters showed that T. r. sonoriana and M. mazzotti could have an important potential role in the prevalence of Chagas disease cases where these species are commonly found. PMID- 16252524 TI - Anopheles triannulatus (Neiva and Pinto): a new Anopheles record from Trinidad, West Indies. AB - We report the first collection of Anopheles triannulatus from Trinidad, West Indies. Adults were captured using human bait while larvae and pupae were collected from a pond located in Valencia, north Trinidad. The associated species was An. oswaldoi. This new record increases the number of mosquito species belonging to the Genus Anopheles to 14 and the total species count from 162 to 163 in Trinidad, West Indies. PMID- 16252525 TI - Introduction and establishment of Aedes (Finlaya) Japonicus japonicus (Theobald) on the island of Hawaii: implications for arbovirus transmission. AB - On November 24, 2003, 1 female adult specimen of Aedes (Finlaya) japonicus japonicus (Theobald) was collected in a New Jersey (NJ) light trap on the island of Hawaii. From June through October, 2004, female and male adults were collected by NJ light traps and gravid traps placed at multiple sites on the island of Hawaii. Larvae were collected in artificial containers and reared to adults for identification. Aedes (Fin.) j. japonicus is the 8th mosquito species to be introduced and established in the State of Hawaii. Currently, this species is known only from the island of Hawaii. Aedes (Fin.) j. japonicus is a competent laboratory vector for a number of arboviruses. Increased quarantine inspections, inspection and treatment of imported used tires and plants, disinsection of airline cargo holds, enhanced vector surveillance, and the development of sanitary corridors around airports and port facilities are necessary to reduce the introduction of vectors and pathogens. PMID- 16252526 TI - First collection of Ochlerotatus japonicus japonicus in the state of Tennessee. AB - This is the 1st report of Ochlerotatus japonicus japonicus in the state of Tennessee, USA. Adults were collected at 5 sites in northwestern Knox County from June 9 through November 3, 2003, and from 1 site in both 2003 and 2004. PMID- 16252527 TI - Identification of Aedes albopictus in urban Nicaragua. AB - Larvae of Aedes albopictus, a mosquito known for transmitting dengue virus, were identified in the city of Leon, Nicaragua, in 2003. Mosquito larvae were collected from a total of 2,225 residences in the 2 largest cities in Nicaragua during the period from June to September of 2003, and larval Ae. albopictus were identified in 4 homes in Leon. This represents the 1st detection of Ae. albopictus in a major Nicaraguan urban center, and increased control efforts appear to have eliminated the mosquito subsequently from Leon. The presence of Ae. albopictus in urban Nicaragua highlights the need for surveillance of areas thought to be free of the mosquito so that early detection and control activities can prevent its spread. PMID- 16252528 TI - Predatory capacity and prey selectivity of nymphs of the dragonfly Pantala hymenaea. AB - Predatory capacity and prey selectivity of nymphs of the dragonfly Pantala hymenaea (Odonata: Libellulidae) were evaluated on larvae of the mosquito Culex quinquefasciatus (Diptera: Culicidae) and larvae of the midge Chironomus plumosus (Diptera: Chironomidae) as prey. With functional response methodology, 7 larval densities were exposed to predator individuals in a glass jar under laboratory conditions. The study was performed in 2 experiments. The 1st was a test system with each prey species alone. The 2nd tested a mixture of both prey species in a 1:1 ratio. Prey selectivity and prey capacity were significantly greater on midge larvae than on mosquito larvae. PMID- 16252530 TI - The only lifetime guarantee is life. PMID- 16252529 TI - Donald James Pletsch--six decades of international commitment. PMID- 16252531 TI - The top 50 prescription medications dispensed in pharmacies in 2004. PMID- 16252532 TI - A literature-based overview of the preparation of teeth. AB - This column has discussed only a few highlights of tooth preparation. Any degree of success is difficult without proper positioning for the operator, patient, and assistant. There are many other aspects to preparation of teeth; these will be discussed in future restorative columns. PMID- 16252534 TI - Digital radiography--should you take the plunge? PMID- 16252533 TI - The dating dentist. PMID- 16252535 TI - Clinical evaluation of indirect resin composite and ceramic onlays over a 24 month period. AB - The purpose of this study was to compare the clinical performance of ceramic and indirect resin composite onlays over a 24-month period. A total of 94 onlay restorations in 47 patients were included in this study. Cavities were prepared, full-arch impressions were taken, and onlays were fabricated in the laboratory. All restorations were placed with a dual-cured luting resin composite system. Restorations were evaluated at baseline and at 6, 12, 18, and 24 months. Differences between Alpha scores were analyzed statistically. The recall rate of this study was 93.6%. At the six-month evaluation, one onlay failed due to pulpitis. Other than the color match, there was no significant difference between indirect resin composite and ceramic onlays (p > 0.05). For indirect resin composites, Alpha score differences pertaining to color match, marginal adaptation, and surface texture were found to be statistically significant (p < 0.05) between the baseline and the 24-month recall. For ceramic restorations, marginal adaptation criteria was the only significant difference over the 24 month period (p < 0.05). At the end of 24 months, both indirect resin composite and ceramic onlays were considered to be successful clinically. PMID- 16252536 TI - Extranodal marginal zone lymphoma: a case report and review of the literature. AB - The World Health Organization (WHO) defines extranodal marginal zone lymphoma (MZL) as an indolent B-cell lymphoma. This disease involves a heterogeneous group of B-cell tumors that occurs primarily within the lymph nodes, spleen, or extranodal tissues. Initial MZL treatment consists of chemotherapy and/or radiotherapy and is associated with a high survival rate when detected early. This article highlights an unusual case of extranodal MZL in the parotid area. PMID- 16252537 TI - Primary oral and perioral amyloidosis associated with multiple myeloma. AB - Three years after being diagnosed with multiple myeloma, a patient sought treatment for swelling on the floor of the mouth, associated with hardening of the soft tissues on the right perioral region, loss of facial expression, and difficulty opening his mouth. The patient reported improvement following an incisional biopsy for microscopic diagnosis. Eighteen months later, the patient showed no clinical alterations. PMID- 16252538 TI - Shear bond strength of direct composite repairs in indirect composite systems. AB - Indirect composite restorations may require repair during clinical service. This study evaluated the shear bond strength of direct composite repairs in indirect composites after four surface treatment procedures. Cylinders of three indirect composites (n = 10 per group) received the following surface treatment: sandpaper finishing (control group), 50 microm aluminum oxide sandblasting, 50 microm aluminum oxide sandblasting (plus 10% fluoridric acid etching), and 30 microm silaned silicon sandblasting. Repair procedure was simulated with the application of an adhesive agent and packing of a direct composite. After storage for one hour in relative humidity and 23 hours in distilled water at 37 degrees C, the specimens were tested for shear bond strength using a universal testing machine at a crosshead speed of 0.5 mm/min. Groups treated with aluminum oxide sandblasting or CoJet Sand had the highest mean values of shear bond strength with no statistically significant difference between the two techniques. PMID- 16252539 TI - Self-report health questionnaire: a necessary and reliable tool in dentistry. AB - This study was designed to assess whether the number and type of medical conditions elicited by self-report on a questionnaire were equivalent to those obtained through verbal inquiry by blinded trained dentists. The study sample consisted of 100 adult patients who were seeking treatment from the University of Kentucky Orofacial Pain Center. Evaluations occurred between September 2003 and January 2004. Patients completed a standardized medical health questionnaire containing 92 questions. Subsequently, the patients were questioned verbally about their medical history by uniformly trained dentists, using a systems review approach as part of routine clinical protocol. The medical histories obtained by both methods were statistically analyzed using Kappa values, paired and independent sample t-tests. The nature of the questionnaire did not distinguish between past and present conditions, and did not give information regarding onset, severity, duration, and impact of the marked conditions. It was concluded that even though a self-reported health questionnaire is precise and consistent, more detailed information can be obtained by verifying marked medical conditions through a verbal inquiry. Every history and examination should include a combination of both a self-reported medical health questionnaire and a verbal inquiry to aid in diagnosis and treatment. PMID- 16252540 TI - The clinical evaluation of a single-bottle adhesive system with three restorative materials in children: six-month results. AB - This study was designed to evaluate the clinical performance of a single-bottle adhesive system with three restorative materials placed in children using the total-etch technique. One hundred thirty restorations were placed on conservative preparations in 36 children using the incremental placement technique in a clinical environment. The restorations were evaluated within a week of placement (baseline) and again after six months. The criteria evaluated included retention, color match, marginal discoloration, marginal integrity, surface roughness, postoperative sensitivity, recurrent caries, and wear. Indirect evaluations of anatomical form, marginal adaptation, and wear also were conducted. Clinical evaluation demonstrated no significant differences between the three restorative materials. Five restorations had to be replaced: two compomer, two packable composite, and one amalgam. There was no clinically detectable wear in any of the restorations. PMID- 16252541 TI - Survey of general dentists regarding posterior restorations, selection criteria, and associated clinical problems. AB - This survey was designed to provide insight concerning the materials that general dentists use for direct and indirect posterior restorations. A stratified random sample of dentists (n = 2,880) from the Academy of General Dentistry's membership list received a questionnaire that elicited information on the clinician's current use of amalgam, direct, and indirect composite materials; reasons for material selection; and reported complications with composite restorations. Data were analyzed using descriptive statistics as well as t-test, Mann-Whitney, and chi square for group comparisons. A total of 714 dentists (26.3%) responded. Direct composite was the material used most commonly for posterior intracoronal restorations. Dentists in amalgam-free practices (31.6%) were significantly more likely (p = 0.001) to use direct composite than dentists whose practices used amalgam. Both groups weighed clinical decision factors and ranked perceived complications differently. Dentists in amalgam-free practices were less likely (p = 0.001) to consider caries rate, patient demand, and moisture control as important clinical decision factors and were more likely to report never experiencing complications. PMID- 16252542 TI - Interradicular radiolucency of maxilla. PMID- 16252543 TI - Ban on sale of non-iodized salt for human consumption: a step in the right direction. PMID- 16252544 TI - Validation of EORTC quality-of-life questionnaire in Indian women with operable breast cancer. AB - BACKGROUND: The European Organization for Research and Treatment of Cancer (EORTC) module QLQ-C30 and the breast cancer-specific module BR-23 have been validated worldwide to assess the quality of life (QOL) in women with breast cancer. No such study has been published on Indian women using EORTC questionnaires. METHODS: QOL was assessed in relation to surgery, adjuvant chemotherapy, radiation therapy and hormone therapy in 299 Indian women with operable breast cancer (OBC) at the Breast Unit of Tata Memorial Hospital (TMH), Mumbai, from October 1998 to September 2001. The QLQ-C30 module was used to assess physical health, emotional, cognitive and social functioning, and the BR 23 module to assess breast cancer treatment-related symptoms. Assessment was done at 3 visits: visit 1 (after surgery); visit 2 (during adjuvant therapy) and visit 3 (on completion of adjuvant therapy). RESULTS: Of the 299 women at first visit, 274 (91.6%) completed the visit 2 questionnaire and 239 (80%) completed the visit 3 questionnaire. Only those women who filled the questionnaires at all 3 visits were included as 'valid visits' for analysis (193 of 299; 64.5%). The reliability and validity of the English and translated versions of the questionnaires were tested by Cronbach alpha (0.61-0.96) and item-scale correlation (0.63-0.93). Women with breast conservation treatment had a superior body image as compared to those with mastectomy (p <0.001). Physical, emotional and cognitive functions were not related to the type of surgery. Global QOL, physical, sexual and role functioning were found to deteriorate with chemotherapy (p < or = 0.01). Radiotherapy had only local adverse effects (p < 0.001 ), while hormone therapy had no adverse impact on QOL. CONCLUSION: QLQ-C30 and BR-23 questionnaires can be used reliably to assess QOL in Indian patients. The translated versions were found to be valid for further use in clinical trials on Indian women with breast cancer. PMID- 16252545 TI - Breast conservation therapy for breast cancer: patient profile and treatment outcome at a tertiary care cancer centre. AB - BACKGROUND: Breast conservation therapy is a well-established treatment modality for early breast cancer. It is not widely practised in developing countries because of a lack of awareness and treatment facilities, and physician and patient bias. We analysed our experience of breast conservation therapy. METHODS: We retrospectively reviewed 102 patients who had undergone breast conservation surgery and axillary dissection for breast cancer. Surgery was followed by 45 Gy of radiation to the whole breast and 15-20 Gy of tumour bed boost. All high risk patients received adjuvant systemic therapy. The disease profile, morbidity and treatment outcome were analysed. RESULTS: Out of 902 patients, 102 underwent breast conservation therapy (90 had early breast cancer and 12 had locally advanced breast cancer). Only 19.6% of patients with early breast cancer received breast conservation therapy. One-third of the patients had had a prior surgical intervention. The mean tumour size was 2.8 cm, 44% had nodal involvement and 29% were oestrogen- and progesterone-receptor negative. At a mean follow up of 32 months, only 1 patient had local recurrence, and the 5-year projected disease free and overall survival were 82% and 88%, respectively. CONCLUSION: Breast conservation therapy should be offered to suitable breast cancer patients. Strict adherence to protocol-based therapy and active multidisciplinary coordination are crucial for a successful breast conservation therapy programme. Education of the patient as well as the physician population is necessary for increasing the breast conservation therapy rates in India. PMID- 16252546 TI - Occult cobalamin and folate deficiency in Indians. AB - BACKGROUND: Our aim was to assess cobalamin and folate levels in normal Indian subjects before undertaking a prospective study of megaloblastic anaemia. METHODS: We took samples from 25 men and 25 women to establish the normal range. The exclusion criteria for subjects were age below 18 years and above 65 years, haemoglobin < 12 g/dl, and those who were pregnant, lactating or on any medication including vitamin supplements. A complete blood count and blood film examination for hypersegmented neutrophils were done. Serum cobalamin and folate assays were performed by a competitive immunoassay. The reference range supplied with the kits for serum cobalamin was 100-700 pg/ml and for serum folate it was 3 22 ng/ml. RESULTS: Since many 'normal' subjects in the sample showed values below the normal reference range, the numbers sampled were increased to 46 men and 50 women. Of all the subjects tested, 46.9% had subnormal values of one of the two vitamins. The normal ranges for serum cobalamin established in this study were- men 100-388 pg/ml and women 105.3-434 pg/ml. Of the 46 men tested, 17 (36.9%) had low cobalamin levels and of the 50 women tested, 23 (46%) had low cobalamin levels. Levels < 50 pg/ml were seen in 46.9% of these subjects. The normal ranges for serum folate in the study were--men 3.1 to > 22 ng/ml, women 3-12.26 ng/ml. In the study group, 8 men (17.3%) and 6 women (12%) had folate deficiency. Eight subjects (17%) had combined deficiency of the two vitamins. The mean corpuscular volume was not informative and was elevated in only 1 subject. Hypersegmentation of neutrophils was present in 75% of deficient subjects. CONCLUSION: We established normal levels for serum cobalamin and folate in our study group. Of the subjects studied, 46.9% had subnormal levels of serum cobalamin or serum folate, cobalamin deficiency being five times more common than folate. Hypersegmentation of neutrophils was a better indicator of occult megaloblastosis than the mean corpuscular volume. PMID- 16252547 TI - In vitro efficacy of ciprofloxacin and gentamicin against a biofilm of Pseudomonas aeruginosa and its free-living forms. AB - BACKGROUND: Infection with Pseudomonas aeruginosa is a major cause of morbidity and mortality in patients with cystic fibrosis (CF). With chronicity of infection, the organism acquires a mucoid phenotype and grows as microcolonies in a biofilm in the respiratory passages of the host. This acts as a protective niche and helps the organism to evade the host immune response. In the biofilm the organism has a high resistance to antibiotics, leading to problems in eradication, and poses a therapeutic challenge. We studied the antimicrobial susceptibility of P. aeruginosa ATCC 27853 in a biofilm and as free-living forms against ciprofloxacin and gentamicin, the commonly used antibiotics in patients with CF. METHODS: Biofilm formation of P. aeruginosa ATCC 27853 was characterized by in vitro biofilm formation assay. The biofilm was detected by light microscopy and quantitated by measuring the absorbance at 575 nm and by viable bacterial counts. After the maximal biofilm was established, the effect of various concentrations of ciprofloxacin (1, 2 and 4 microg/ml) and gentamicin (4, 8, 16, 32 and 64 microg/ml) was observed and the minimal biofilm eradication concentration (MBEC) determined. The minimal bactericidal concentration (MBC) of both the antibiotics was determined against the free-living forms of the organism. The MBEC of the two antibiotics was further compared with the MBC. RESULTS: On microscopic examination, the maximal biofilm of P. aeruginosa was established on a coverslip at 12 hours, the maximum absorbance was at 575 nm and viable counts were observed at 12 hours, which corresponded to the maximal biofilm production. The organisms in the biofilm showed a 4-fold greater resistance against ciprofloxacin and gentamicin as compared to the free-living forms. CONCLUSION: In biofilm, P. aeruginosa shows greater resistance against antibiotics. This renders these antibiotics ineffective, leading to chronic and persistent infections. PMID- 16252548 TI - Prenatal diagnosis of von Willebrand disease in a family. AB - We report the successful prenatal diagnosis of von Willebrand disease (VWD) in a family with type 3 severe VWD by the indirect method of gene tracking using polymorphic markers of intron 40 of the von Willebrand factor (VWF) gene. The couple had a daughter diagnosed to have type 3 VWD. Chorionic villus sampling (CVS) was done in the eleventh week of gestation of a subsequent pregnancy. The 3 VNTR polymorphic markers VWF1, VWF2 and VWF3 of intron 40 of the VWF gene were used for linkage studies. DNA in the affected VWD patient, the father and mother as well as in the CVS using VWF1 and VWF3 polymorphic markers revealed that the foetus was affected. The family chose to abort the foetus. In a subsequent pregnancy, similar investigation revealed a normal foetus. Prenatal diagnosis in families with a diagnosed case of VWD can be used to determine the status of the foetus. The technique is inexpensive. PMID- 16252549 TI - Infections transmitted from donors to recipients following organ transplantation. AB - There is an ever-increasing gap between the number of donors and those waiting for organ grafts, resulting in increased waiting times and mortality on transplant waiting lists. Consequently, every potential donor must be considered for possible transplantation even if they are outside the conventional donor criteria. To address this imbalance, organs are currently transplanted from living donors, older donors, haemodynamically unstable and non-heart-beating donors, and donors with prior infections. There is a potential to transmit infections and, to a lesser extent, malignancy from the donor organ to the immunosuppressed recipient, and this may also have an effect on subsequent organ function in the recipient. Thus, transmission of infections from organ donors to recipients represents low but serious potential risks that must be weighed against a candidate's risk of dying before a transplant becomes available. PMID- 16252550 TI - Early prostate cancer: radical prostatectomy or watchful waiting. PMID- 16252552 TI - Killing for the state: death penalty and the medical profession: a call for action in India. PMID- 16252551 TI - The Tribal Health Initiative model for healthcare delivery: a clinical and epidemiological approach. AB - BACKGROUND: Tribal populations generally have poor health outcomes, often because of a healthcare delivery system that does not cater to their needs. This study evaluates a current healthcare model for tribals, and explores it in combination with the health status of the target population, placing emphasis on the long term sustainability and cross-implementation of the model. METHODS: We examined the health system from the perspective of the base hospital, by concentrating on mortality patterns, inpatient incidence of selected infectious and non-infectious illnesses, and the preventive and curative health services administered by the hospital to the community. RESULTS: Gender susceptibility patterns revealed disparities in anaemia and tuberculosis besides fluctuations in gastrointestinal disorders, tuberculosis and typhoid. A combination of gender- and age susceptibility patterns revealed specific age intervals for mental health-related disorders. Mortality patterns indicated an Increase in youth deaths and suicide, with an overall reduction in infant mortality. However, an increased tribal confidence in allopathic medicine was noted after implementation of the health system. CONCLUSION: The base tribal hospital is important in administering primary and secondary healthcare, health education, disease surveillance, community outreach and for continued confidence in allopathic medicine. Diet based morbidities may be combated via organic farming and banning local alcohol production, while anaemia may be combated through continued iron, salt and folic acid supplementation to women. The formulation of mental health programmes and long term educational initiatives at the village level are critical to reducing suicide and infant mortality. Further epidemiological studies are required to gain a complete picture of health within the population, and successful implementation of the model elsewhere must factor in sociocultural disparities among tribes. PMID- 16252553 TI - Ethical issues in international research, Harvard School of Public Health, Boston, USA, 13-17 June 2005. PMID- 16252555 TI - The great downpour: lessons still to be learnt. PMID- 16252554 TI - Generic drugs. PMID- 16252556 TI - A few words. PMID- 16252557 TI - The Kentucky Medical Association report on the use of electronic medical records by Kentucky physicians. PMID- 16252558 TI - Daniel W. Varga, MD KMA President 2005-2006. PMID- 16252559 TI - Hernation of the heart following radical pneumonectomy: report of a case. PMID- 16252560 TI - Rural Kentucky's physician shortage: strategies for producing, recruiting, and retaining primary care providers within a medically underserved region. PMID- 16252561 TI - August. PMID- 16252563 TI - Rules that work when unions try to organize. PMID- 16252562 TI - Breast Cancer Awareness month. PMID- 16252564 TI - How do empowered patients affect hospitals? PMID- 16252565 TI - [Usefulness of lung and right ventricular thallium-201 uptake during single photon emission computed tomography in exercise testing of patients with coronary artery disease]. AB - OBJECTIVES: Increased pulmonary or right ventricular 201Tl uptake during the exercise test has been used as a marker of multivessel coronary artery disease. The most useful method for assessing the severity of coronary artery disease was evaluated among conventional evaluation of single photon emission computed tomography (SPECT), measurement of lung to heart uptake ratio (L/H), and right ventricular to left ventricular uptake ratio (RV/LV) on 201Tl images during exercise testing. METHODS: Regions-of-interest (4 X 4 pixels) were placed at the lung and the heart, and L/H was defined as mean lung uptake/mean heart uptake. Correspondingly, regions-of-interest (4 X 4 pixels) were placed at the RV and the LV, and RV/LV was defined as maximum RV uptake /maximum LV uptake. L/H and RV/LV on the initial image were analyzed in 216 patients(angiographically normal coronary arteries: 89, single-vessel disease: 82, multivessel disease: 45). The diagnostic value was evaluated using the receiver operating characteristic curve. RESULTS: All methods showed significantly higher values in patients with multivessel disease than in patients with no coronary artery disease or single vessel disease. L/H was significantly higher in patients with prior myocardial infarction and RV/LV was significantly higher in patients without infarction. The sensitivity of only conventional SPECT evaluation for multivessel coronary artery disease was low (sensitivity 53%, specificity 94%). However, addition of evaluation of L/H and RV/LV to SPECT improved the sensitivity for multivessel coronary artery disease (sensitivity 93%, specificity 49%). CONCLUSIONS: The diagnostic sensitivity for multivessel coronary artery disease was improved by adding L/H and RV/LV to conventional evaluation of exercise 201Tl SPECT. L/H and RV/LV during exercise 201Tl imaging may provide additional information regarding the severity of coronary artery disease. PMID- 16252566 TI - [Exercise-induced ST-segment depression and myocardial ischemia in patients with hypertrophic cardiomyopathy: myocardial scintigraphic study]. AB - OBJECTIVES: Patients with hypertrophic cardiomyopathy (HCM) sometimes develop myocardial ischemia during exercise in the absence of coronary lesions. The relationship between myocardial ischemia and ST-segment depression was investigated during exercise testing in patients with HCM. METHODS: Regional hypoperfusion and/or transient left ventricular cavity dilation, a parameter of subendocardial hypoperfusion, were assessed on exercise 99mTc-tetrofosmin myocardial scintigraphy in 42 patients with non-obstructive HCM. The scintigraphic results were further correlated with the ST-segment responses to exercise. RESULTS: Regional hypoperfusion or transient left ventricular cavity dilation were observed in 19 (45%) or 16(38%)patients with HCM, respectively. The incidence of ST-segment depression > or = 0.1 mV during exercise testing was similar in HCM patients with regional hypoperfusion, with transient left ventricular cavity dilation, and without hypoperfusion (42%, 38%, 38%, p = 0.95). Furthermore, exercise-induced ST-segment depression > or = 0.1 mV occurred similarly irrespective of symptoms, exercise tolerance, the degree or the site of hypertrophy, or the presence or absence of resting ST-segment depression. CONCLUSIONS: ST-segment depression during exercise testing was common in patients with HCM, but seems to be an unreliable marker of myocardial ischemia as assessed by exercise scintigraphy. PMID- 16252568 TI - [Acute myocardial infarction with variable clinical manifestations: probable catastrophic primary antiphospholipid antibody syndrome: a case report]. AB - A 62-year-old diabetic man was admitted to our hospital because of acute myocardial infarction. Emergent coronary angiography showed multiple thromboembolic occlusions in the distal circumflex and anterior descending arteries. For the first 2 weeks of hospitalization, he suffered multiple organ manifestations including the gastrointestinal, central nervous, renal and respiratory systems. The anticardiolipin beta2GP1 complex antibody titer on the 15th day was as high as 27.2 U/l (normal value < 3.5). These clinical manifestations and laboratory findings suggested catastrophic antiphospholipid antibody syndrome. He was discharged on the 83rd day with anticoagulant therapy and regular hemodialysis. Acute myocardial infarction is rare as the initial manifestation of catastrophic antiphospholipid antibody syndrome. PMID- 16252569 TI - Beneficial effects of human atrial natriuretic peptide on renal function in a patient with repeated coronary interventions: a case report. AB - Among percutaneous coronary intervention post procedural complications, renal acute dysfunction due to administration of contrast agent is commonly seen in patients with pre-existing renal impairment, especially with co-existent diabetes mellitus and/or congestive heart failure. Although several strategies have been proposed to ameliorate this condition, their effects are inconsistent. A 66-year old diabetic man at high risk for this complication underwent three distinct percutaneous coronary intervention procedures. Different strategies were used during each intervention to prevent the development of contrast medium-induced nephropathy. Dramatic renal protection was observed with human atrial natriuretic peptide administration or hemofiltration, whereas saline hydration had no apparent effect. PMID- 16252570 TI - [Abnormal Doppler flow pattern from the supraclavicular fossa observed in the left internal thoracic artery bypass graft]. PMID- 16252567 TI - Gorlin formulas for aortic regurgitant and mitral regurgitant orifice areas: experimental studies. AB - BACKGROUND AND OBJECTIVES: The measurement of aortic regurgitant and mitral regurgitant orifice areas has recently been pioneered. The present study tried to establish Gorlin formulas for aortic regurgitant and mitral regurgitant orifice areas for evaluating the severity of aortic regurgitation and mitral regurgitation. METHODS: Seventeen stable hemodynamic states for mitral regurgitation and 22 for aortic regurgitation were studied in sheep. Aortic regurgitant and mitral regurgitant orifice areas were determined by dividing regurgitant volume per second by the time integrals of the continuous aortic regurgitant and mitral regurgitant wave velocities. Aortic regurgitant and mitral regurgitant orifice areas calculated by the formulas using echocardiographic data were compared with other measurements by electromagnetic flowmeters. RESULTS: The Gorlin formulas were aortic regurgitant orifice area = per second valve flow/ 50.7 square root mean pressure gradient, and mitral regurgitant orifice area = per second valve flow/ 27.5 square root mean pressure gradient. Simple linear analysis between aortic regurgitant fractions, regurgitant volume per beat and peak aortic regurgitant flow rates, and aortic regurgitant orifice areas derived from the formula showed moderately good relationships (r = 0.73, 0.81 and 0.83). Mitral regurgitant orifice areas calculated by the formula also correlated with mitral regurgitant fractions, regurgitant volume per beat and peak mitral regurgitant flow rates (r = 0.86, 0.92 and 0.74). CONCLUSIONS: The Gorlin formulas for aortic regurgitant and mitral regurgitant orifice area may provide an index for evaluating the severity of regurgitation. PMID- 16252571 TI - Imported drug-resistant severe malaria. PMID- 16252572 TI - Gestational trophoblastic disease: our patients deserve better case detection and follow up. PMID- 16252573 TI - Efficacy of rivastigmine on activities of daily living in Sri Lankan patients with Alzheimer disease and on improving caregiver burden: a prospective study. AB - OBJECTIVE: This open label, parallel group, prospective cohort study investigated the efficacy of rivastigmine treatment on activities of daily living (ADL) in patients with mild to moderate Alzheimer's disease (AD) and the possible benefits of this therapy on caregiver stress levels. METHODS: Thirty eight consecutive patients with mild to moderate AD were recruited; 22 received rivastigmine 3-6 mg twice daily (treatment group) for 20 weeks. Sixteen patients who did not receive rivastigmine served as the control group. The 17-item ADL Index was used to assess ADL and to determine the presence of functional deterioration. Caregivers were evaluated with the Caregiver Stress Scale (CSS). Each patient was required to have a committed caregiver and all caregivers were interviewed and administered the ADL Index and the Caregiver Stress Scale (CSS) at the start of treatment (week 0) and at the end of 20 weeks of treatment (week 20). RESULTS: Patients in the control group showed a significant decline in ADL Index score at 20 weeks compared to rivastigmine-treated patients (difference in mean ADL Index score = 8.5; p < 0.001). At week 20, mean change from baseline scores for CSS total and individual domain scores were better for caregivers in the treatment group than those in the control group (CSS total mean difference = 19.2). CONCLUSION: We conclude that treatment of AD patients with rivastigmine for 20 weeks produces a significant improvement in patient ADL functioning, and lower levels of caregiver stress. PMID- 16252574 TI - Morbidity pattern and household cost of hospitalisation for non-communicable diseases (NCDs): a cross-sectional study at tertiary care level. AB - OBJECTIVE: To determine the pattern of morbidity and the demographic and socioeconomic characteristics of patients seeking in-patient services for noncommunicable diseases (NCDs) in medical units of a tertiary care hospital, and to estimate the economic burden imposed by these admissions on the households. METHODS: A descriptive cross-sectional study was conducted in medical units of the Colombo North Teaching Hospital, Ragama. Data were collected using a pre tested interviewer-administered questionnaire. Morbidity patterns and demographic and socio-economic characteristics of patients with NCDs were determined. Direct and indirect components of the household cost of hospital stay were estimated. RESULTS: Fifty five per cent of the patients men male and the largest age group (11%) was 50-54 years. Seventy per cent were above 40 years of age, and 63% represented social classes 4 and 5. Diseases of the circulatory system were the commonest (31%). Median household cost of the total hospital stay was Rs. 852.00 (inter-quartile range Rs. 351.00-1885.00) of which 70% were direct costs. Median daily cost was Rs. 340.00 (interquartile range Rs.165.00-666.00). Only 44% of patients incurred an indirect cost. Cost of travelling was the main contributor (36%) to the household cost. Laboratory investigations contributed 16%. CONCLUSIONS: Most patients seeking in-patient services were from a poor socioeconomic background. The economic burden imposed by the admission to the household was mainly due to direct costs incurred for travelling and investigations. PMID- 16252575 TI - Clinical, biochemical and histological characteristics of a Sri Lankan population of non-alcoholic steatohepatitis (NASH) patients. AB - BACKGROUND: Non-alcoholic steatohepatitis (NASH) is common and can progress to cirrhosis. It has been regarded as a 'disease of affluence' and there are only a few reports from developing countries. OBJECTIVE: To describe the clinical, biochemical, and histological characteristics of a cohort of NASH patients in Sri Lanka, and to determine their short term outcome following modifications of lifestyle. METHOD: Patients who had a liver biopsy for investigation of raised hepatic enzymes were assessed during the period May 1999 - May 2003. Patients who had an alcohol intake of over 40 g/week were excluded. Detailed clinical and biochemical data of patients with histologically confirmed NASH were compiled. Histological grading and staging was done using the Brunt system. The patients were advised on lifestyle modifications and the control of diseases known to be associated with NASH. They were followed up at 3 -monthly intervals. RESULTS: During the study period liver biopsies were performed on 296 patients and 100 ( 35.1%) were diagnosed as having NASH. (Men = 79, Mean age 37.2 years, SD 10.6). Risk factors for NASH included diabetes mellitus (55%), obesity (52%), hyperlipidaemia (54%), a family history of risk factors (66%) and a high dietary fat intake (66%). However, 44.3% of men and 33.3% of women were not overweight. The histological grading and staging of 80 biopsies showed Grade 1 in 31 (38.8%), Grade 2 in 29 (36.3%), Grade 3 in 20 (25%), Stage 1 in 57 (71.3%) Stage 2 in 13 (16.3%), Stage 3 in 2 (2.5%) and Stage 4 in 8 (10%). In 55/91 (60.4%) patients who were followed up for a median of 2.5 years (range 1-4 years) the serum transaminases returned to normal in a median of 7 months (range 3-14 months). CONCLUSION: The clinical, biochemical and histological features of NASH patients in our series are similar to that reported in western countries. However "lean males" accounted for a significant proportion. In the short term a majority of patients showed improvement in serum transaminases with lifestyle modification. PMID- 16252576 TI - Adverse effects of teenage pregnancy. AB - RATIONALE: Recent studies have suggested that teenage pregnancies are not as hazardous as thought to be earlier. OBJECTIVE: To compare the sociodemographic data, obstetric complications and attitudes towards family planning in teenagers and older women. DESIGN AND SETTING: A prospective cohort study at the University Obstetrics Unit, Teaching Hospital, Galle. SUBJECTS AND METHOD: Sociodemographic data, details of antenatal care and family support, antenatal complications, gestation at delivery, mode of delivery, the proportion of unplanned pregnancies, and the possible effects of contraceptive counselling, in two groups of pregnant teenagers (13-16 years, n = 95 and 17-19 years, n = 250) were compared with a control group of pregnant women (20-24 years, n = 275). RESULTS: The teenagers were from lower socioeconomic strata and the younger teenagers were significantly less educated than the controls. Teenagers had a significantly higher risk of anaemia (Odds Ratio (OR) = 2.3, 95%CI = 1.7-3.3, p < 0.001). The younger teenagers had a significantly higher risk of gestational hypertension (OR = 4.8, 95%CI = 1.8-13.0, p < 0.001) and pre-eclampsia (OR = 5.0, 95%CI = 1-27, p = 0.03). The older teenagers had a significantly higher risk of delivery before 34 weeks of gestation (OR = 13.6, 95%CI = 1.8-287, p = 0.001). There were no significant differences in the mode of delivery. The younger teenagers had a much higher proportion (54%) of unplanned pregnancies compared to the controls (16 %). A significantly higher proportion of younger teenagers (48%) and older teenagers (25 %), if counselled, would have delayed their pregnancies compared to the controls (10 %). CONCLUSION: Teenage pregnancies, especially those below 17 years of age have a significantly higher risk of adverse outcomes. A large proportion of these pregnancies is unplanned and could be prevented by counselling. PMID- 16252577 TI - Picture quiz 1. Miliary tuberculosis. PMID- 16252579 TI - Mirror writing in a patient with Alzheimer disease. PMID- 16252578 TI - Imported quinine resistant Plasmodium falciparum malaria in Sri Lanka. PMID- 16252580 TI - A case of hereditary spastic paraplegia with demyelinating polyneuropathy. PMID- 16252581 TI - An unusual cause of haematuria. PMID- 16252582 TI - Perspective of violence and crime. PMID- 16252583 TI - Enteric fever: do elevated liver enzymes influence outcome? PMID- 16252584 TI - Variations in the presentation of sickle cell beta thalassaemia--a report of two cases. PMID- 16252585 TI - Metanephric adenoma mimicking renal cell carcinoma. PMID- 16252586 TI - Cyclical intravenous pamidronate therapy in children with osteogenesis imperfecta. PMID- 16252587 TI - Use and interpretation of phrases in histopathology reports: interdisciplinary meetings will help to reduce the confusion. PMID- 16252588 TI - Nurse strikes bring nation-wide attention; 'like Samuel Gompers' says Wall Street Journal. PMID- 16252590 TI - ANA report 'challenge of today'. PMID- 16252591 TI - Have nurses changed? PMID- 16252592 TI - Nursing: a service to the public. PMID- 16252593 TI - Nursing and ethics. PMID- 16252594 TI - Professional culpability and cerebral palsy. PMID- 16252595 TI - False antiquities: authentication of art & archaeological objects--the Turin Shroud & the Vinland Map. PMID- 16252596 TI - Murder by insulin. PMID- 16252597 TI - Medical negligence in Italy: the criminal inquiry. AB - In the past decade, there has been an exponential increase in legal proceedings against doctors due to allegations of medical negligence. The via crucis of the doctor under investigation for alleged medical negligence in Italy is little known beyond the Alps. The authors hope this paper will bring to the attention of medical and legal colleagues this complex issue of the process followed by the justice machinery in Italy where the criminal route in pursuit of justice is the norm. PMID- 16252598 TI - No compensation for unwanted healthy child. PMID- 16252599 TI - The consequences upon victims of violent crime including domestic violence. PMID- 16252600 TI - Legal aspects of renal failure. PMID- 16252601 TI - Is there any future for inquests and inquiries? PMID- 16252602 TI - The doctrine of double effect and the law of murder. PMID- 16252603 TI - Parental alienation and the judiciary. PMID- 16252604 TI - Court orders HIV-1 test for baby. PMID- 16252605 TI - From the Association of Lawyers for the Defence of the Unborn. PMID- 16252606 TI - Muriel Driver Memorial Lecture. Participation and occupation. PMID- 16252607 TI - Evaluation of an occupational therapy mentorship program. AB - BACKGROUND: It was identified that there is a lack of evidence for the effectiveness of formal mentorship programs, no evidence supporting mentor groups and little research in mentorship specific to occupational therapy. PURPOSE: The following quantitative study evaluates the mentorship program offered within the occupational therapy curriculum at The University of Western Ontario. METHODS: One hundred and seventy nine surveys were analyzed to evaluate participants' perceptions of the program's strengths, weaknesses, changes needed, effectiveness, and whether it should continue to be offered. RESULTS: In general, mentors and mentees reported that the mentorship program should continue to be offered and that it was effective in developing professionalism. However, the structure of the program, clarification of its requirements, and more support for the mentors were identified as some areas for change. IMPLICATIONS: Results can impact further program development at this university and other occupational therapy programs which may have a similar program or are interested in developing a mentorship program. PMID- 16252608 TI - ADL differences in individuals with unilateral hemispheric stroke. AB - BACKGROUND: Literature regarding the ability of individuals who have a cerebrovascular accident (CVA) to perform activities of daily living (ADL) is inconclusive regarding the impact of gender, age or side of the lesion. PURPOSE: To determine if people with a CVA differ in their abilities to perform ADL tasks and actions as affected by their gender, age, and side of the lesion. METHOD: A descriptive comparison of 3878 people with a right or left CVA included in the Assessment of Motor and Process Skills (AMPS) database. RESULTS: People with stroke demonstrated statistically significant gender, age, and side of CVA differences in overall ADL ability. However, the gender and side of CVA differences were not clinically detectable. Increased age was associated with a gradual decline in ADL ability. CONCLUSION: Individuals with a right or left CVA have similar abilities when performing ADL tasks and actions. PRACTICE IMPLICATIONS: These findings indicate that occupation-based intervention, which focuses on the utilization of intact ADL skills to compensate for ADL skill deficits (vs. the utilization of tests of body function), may be a more efficient and effective means of planning and implementing occupational therapy intervention for individuals with a stroke. PMID- 16252609 TI - [Perceived self-efficacy is associated with social participation in adults with traumatic brain injury]. AB - BACKGROUND: Identification of the factors facilitating the social participation of adults who have sustained a traumatic brain injury can help occupational therapists with the direction for their interventions. Earlier studies centered on identifying the socio-demographic characteristics and the disabilities associated with social participation. PURPOSE: The aim of this study was to examine the association between perceived self-efficacy, a positive concept derived from social cognitive theory and social participation. METHODS: A cross sectional and correlational research design was used with 53 adults who sustained a traumatic brain injury between 1995 and 2000 and lived in their natural environment. Two measuring tools were used: a self-administered questionnaire evaluating the perceived self-efficacy and a questionnaire evaluating social participation, administered by an examiner through a face-to-face interview. RESULTS: The results indicate that the perceived self-efficacy explains 40% of the variance of the social participation. PRACTICE IMPLICATIONS: This association suggests that social cognitive theory can constitute a reference model for occupational therapists working with this clientele. PMID- 16252610 TI - Continuing professional education and the Internet: views of Alberta occupational therapists. AB - BACKGROUND: Occupational therapists have identified barriers to accessing continuing professional education (CPE) in the traditional face-to-face formats. One alternative to traditional, centrally located, face-to-face CPE is course delivery through the Internet. PURPOSE: This study examined Alberta occupational therapists' perceptions of Internet-based CPE. METHOD: A questionnaire was mailed to 800 randomly sampled Alberta occupational therapists (response rate = 35.5%; n = 281). RESULTS: Respondents pursued CPE to increase skills, knowledge and maintain clinical competency. They reported that a face-to-face CPE course was more useful than distance courses. Although almost 90% of respondents had access to computers with an Internet connection at home or at work, and nearly 65% thought that their computer knowledge was sufficient, only 2.9% had previously taken Internet-based CPE. PRACTICE IMPLICATIONS: In order for the Internet to be accepted as a common, useful and alternative delivery tool for CPE in occupational therapy, the perceived barriers such as the lack of personal time, cost, and limited interaction with other learners and instructors will need to be addressed. PMID- 16252611 TI - Stroke prevention, blood cholesterol and statins. AB - Statins have a good overall safety profile to date, with no increase in haemorrhagic stroke or cancer. They have favourable effects in the primary prevention of cardiovascular disease in high-risk young as well as elderly populations. Statins reduce the incidence of stroke in high-risk populations (mainly CHD patients, diabetics and hypertensives) even with a normal baseline blood cholesterol level, which argues for a global cardiovascular risk-based treatment strategy. As for CHD, stroke reduction was mainly observed in studies with large between-group LDL cholesterol difference. In patients with prior strokes, statins reduce the incidence of coronary events, but it is not yet proven that they actually reduce the incidence of recurrent strokes in secondary prevention. From a practical point of view, since there was a favourable treatment effect overall in stroke and TIA patients in HPS, it seems reasonable to treat stroke patients with a statin and total cholesterol >135 mg/dL (3.5 mmol/dL). On-going research is aiming to refine patient selection. As anticipated by current US recommendations, patients who are likely to benefit most are those with carotid atherosclerosis, diabetes mellitus, previous coronary heart disease, hypertension, hypercholesterolaemia, or cigarette smoking and LDL cholesterol > 100 mg/dL. PMID- 16252612 TI - Neurological manifestations in severe acute respiratory syndrome. AB - During the worldwide outbreak of severe acute respiratory syndrome (SARS) in 2002 2003, there were 664 probable SARS patients reported in Taiwan. SARS patients usually present with symptoms related to the respiratory system while neurological manifestations have rarely been described. There were three patients who developed axonopathic polyneuropathy 3-4 weeks after onset of SARS; their clinical condition and electrophysiological studies revealed obvious improvement at follow-up. Two SARS patients have experienced myopathy and three other patients developed rhabdomyolysis. These neuromuscular disorders in SARS patients were considered as critical illness neuropathy and myopathy, but the possibility of direct attack by SARS coronavirus on the nerve and muscle could not be excluded. Large artery ischemic stroke were described in five SARS patients with poor prognosis. Multiple factors contributed to this vascular insult included hypercoagulabe status related to both SARS coronavirous and the usage of intravenous immunoglobulin, septic and cardiogenic shock, and possible vasculitis. The relationship between SARS and above neurological problems still needs further clarification. Pathological and microbiological studies are mandatory to delineate this issue. PMID- 16252613 TI - Homozygous deletion genotype of angiotensin converting enzyme confers protection against migraine in man. AB - Studies have shown that migraine may have a major genetic component. Meanwhile, angiotensin converting enzyme (ACE) gene has been implicated as a genetic factor associated with migraine. We designed a case-control study to investigate the association between ACE and migraine in 240 migraine patients and 200 healthy controls, matched by age and sex. There was no significant difference in allelic frequency (I and D) and genotype polymorphism (DD, DI and II) of the ACE gene in migraine patients and controls. Analysis of the difference in ACE polymorphism stratified by gender revealed that male migraine patients with the homozygote DD genotype (ACE-DD) were significantly fewer than that of male controls (OR = 0.331, p = 0.045). There was no existence of a difference among the frequency and duration of headache in each subgroup of migraine patients stratified by ACE genotype. Our findings indicate that ACE-DD may have a slight protective effect against migraine in male patients. PMID- 16252614 TI - The role of emergency medical services in stroke: a hospital-based study in Kaohsiung, Taiwan. AB - PURPOSE: Emergency medical services (EMS) play an important role in acute stroke therapy. The goal of this study was to investigate the roles and determinants of EMS in stroke in Southern Taiwan, Kaohsiung. METHODS: We enrolled stroke patients who arrived at emergent department (ED) of the study hospital within 48 hours after the onset of symptoms. Patients were categorized into arriving by EMS or not EMS. Potential determinants of EMS use for stroke were examined by multivariable analyses and the role of EMS in stroke was discussed. RESULTS: Among 197 stroke patients enrolled, only 44 (22%) patients arrived by EMS at ED. Multivariable logistic regression analysis revealed that the determinants of EMS use were stroke severity measured by NIHSS and non-family member who decided to seek help. Using EMS or not was not associated with the earlier presentation after stroke within 2 hours after attack. CONCLUSION: EMS use was far from sufficient. The transportation time was not the major component of prehospital delay. Both EMS and other vehicles provided prompt delivery. The public should rush to ED either by EMS or other transportation modes when stroke occurred unless use of EMS is proved to provide better outcome in stroke patient in the future study. PMID- 16252615 TI - Adult Acinetobacter meningitis and its comparison with non-Acinetobacter gram negative bacterial meningitis. AB - Between January 1999 and December 2003, 81 cases of single pathogen-related culture-proven Gram-negative adult bacterial meningitis were identified at Chang Gung Memorial Hospital-Kaohsiung. Of these 81 cases, Acinetobacter infection was found in 13 cases. Clinical and laboratory data of these Acinetobacter meningitis patients were studied and were compared with those of other 68 non-Acinetobacter Gram-negative bacterial meningitis (GNBM) patients. Of the 13 implicated Acinetobacter strains, A. baumannii was the most common (12), and the other was A. lwoffii (1). Eleven of these 13 cases were due to a post-neurosurgical infection. The results of the antibiotic susceptibility test of the 13 Acinetobacter strains from cerebrospinal fluid included ceftriaxone, (1/13, 8%), ciprofloaxin (6/13, 46%), ceftazidime (6/13, 46%), cefepime (7/13, 54%), ampicillin-subtactam (7/13, 54%), imipenem (12/13, 92%) and meropenem (12/13, 92%). One strain with pan-drug resistant A. baumannii (PDRAB) emerged in 2003. A statistically significant difference between Acinetobacter meningitis and non Acinetobacter GNBM included hydrocephalus and ceftazidime-resistance. A mortality rate was 30% (4/13), and 7 of the other 9 survivals had severe neurologic deficits. The emergence of Acinetobacter infections in adult post-neurosurgical infections, multiple antibiotic resistant characteristics, and the emergence of PDRAB strain remained a challenge of the initial management of this specific meningitis. Use of carbapenem, especially meropenem, could be considered as one of the initial empiric antibiotics chosen for the management of adult post neurosurgical meningitis. PMID- 16252616 TI - Understanding of senile dementia by children and adolescents: why grandma can't remember me? AB - BACKGROUND: The present study sought to determine the knowledge and attitudes concerning senile dementia among children and adolescents in Taiwan with the intent of using the results in the design of an educational program about senile dementia for this population. METHODS: A 10-item questionnaire was distributed to all students aged 10 to 15 at 7 geographically selected public schools in Taiwan. The teachers in these schools and a group of 20 health professionals who were specialists in dementia were also invited to complete questionnaire for comparison. RESULTS: The majority of children, adolescents and teachers worried that their parents and grandparents might develop dementia in the future. Seven percent of all the student respondents felt that they would be embarrassed to invite classmates to their homes either because of actual demented family members living at home or if the hypothetical situation arose in which they actually had demented family members living at home. The younger students were more likely to have this feeling (chi square for linear trend = 73.636, df = 1, p = 0.000). CONCLUSION: Younger children were more likely to feel stigmatized by dementia in a family member which suggests that they may need more psychological support in order to effectively deal with dementia related issues. These results have important implications for the development of suitable educational programs about dementia in the curricula of primary and junior high schools. PMID- 16252617 TI - Rhabdomyolysis induced by fenoverine: a case report and literature review. AB - Fenoverine is a derivative of phenothiazine. It is commonly used in the treatment of gastrointestinal and gynecological spasmodic disorders. Myalgia is a common side effect, but rhabdomyolysis has only been scarcely reported before. A 77-year old patient without previous history of liver diseases received fenoverine therapy for four days due to abdominal pain. Acute onset of myalgia, proximal muscle weakness and high creatinine phosphokinase (CK) occurred. The foregoing symptoms and signs and abnormal biochemistry improved gradually after discontinuation of fenoverine use. The pathophysiology of fenoverine-induced rhabdomyolysis is unclear. Some predisposing factors, especially liver cirrhosis, had been reported. However, our patient had none of the well-known precipitating factors. Physicians should be aware of the possibility of rhabdomyolysis in patients receiving fenoverine, whether they are healthy or have musculoskeletal or liver dysfunction. PMID- 16252618 TI - Claude's syndrome associated with supranuclear horizontal gaze palsy caused by dorsomedial midbrain infarction. AB - Claude's syndrome caused by dorsal midbrain lesion is characterized by ipsilateral third nerve palsy and contralateral ataxia. To date, reports in the literature concerning Claude's syndrome associated with the midbrain paresis of horizontal gaze are rare. A 62-year-old man suddenly developed left third cranial nerve palsy, right lateral gaze palsy, and right ataxia. Intact Bell's phenomenon and preserved right horizontal oculocephalic reflex suggested the lateral gaze palsy in the right eye was supranuclear in nature. Magnetic resonance imaging (MRI) revealed an infarction in the left dorsomedial midbrain. Although the red nucleus has often been suggested as the lesion site responsible for Claude's syndrome, a lesion of the superior cerebellar peduncle just below and medial to the red nucleus could be responsible for this syndrome. This case demonstrates neurological heterogeneity of midbrain infarction. PMID- 16252619 TI - [EEG manifestations in metabolic encephalopathy]. AB - Normal brain function depends on normal neuronal metabolism, which is closely related to systemic homeostasis of metabolites, such as glucose, electrolytes, amino acids and ammonia. "Metabolic encephalopathy" indicates diffuse brain dysfunction caused by various systemic derangements. Electroencephalogram (EEG) is widely used to evaluate metabolic encephalopathy since 1937, when Berger first observed slow brain activity induced by hypoglycemia. EEG is most useful in differentiating organic from psychiatric conditions, identifying epileptogenicity, and providing information about the degree of cortical or subcortical dysfunction. In metabolic encephalopathy, EEG evolution generally correlates well with the severity of encephalopathy. However, EEG has little specificity in differentiating etiologies in metabolic encephalopathy. For example, though triphasic waves are most frequently mentioned in hepatic encephalopathy, they can also be seen in uremic encephalopathy, or even in aged psychiatric patients treated with lithium. Spike-and-waves may appear in hyper- or hypo-glycemia, uremic encephalopathy, or vitamin deficiencies, etc. Common principles of EEG changes in metabolic encephalopathy are (1) varied degrees of slowing, (2) assorted mixtures of epileptic discharge, (3) high incidence of triphasic waves, and (4), as a rule, reversibility after treatment of underlying causes. There are some exceptions to the above descriptions in specific metabolic disorders and EEG manifestations are highly individualized. PMID- 16252620 TI - Opalski syndrome. PMID- 16252621 TI - Surveillance behavior and prophylactic surgery after predictive testing for hereditary breast/ovarian cancer. AB - This article describes breast or ovarian cancer surveillance practices and prophylactic surgery involving 34 carriers and 34 noncarriers of a BRCA1/2 mutation within the year after predictive testing. It also evaluates the effect of the predictive test result on cancer screening practices and provides insight into factors important in the decision-making process about health-related behavior. Within the year following predictive testing, 9% (3 of 34) of the carriers decided to have a prophylactic mastectomy. The majority of the carriers was adherent to recommendations regarding regular cancer surveillance following predictive testing. Furthermore, carriers' adherence to clinical breast examination and mammography recommendations significantly increased from pre- to posttest and was significantly higher than noncarriers' utilization after testing. Of the carriers eligible for prophylactic salpingo-oophorectomy, 75% had this operation. All carriers who were advised to have regular surveillance of the ovaries had ovarian ultrasounds. The authors gave major attention to factors playing a part in the decision-making process about health-related behavior. PMID- 16252622 TI - The relationship between self-efficacy and self-reported physical functioning in chronic obstructive pulmonary disease and chronic heart failure. AB - In this study, the authors investigated whether self-reported physical functioning of patients with chronic obstructive pulmonary disease (COPD) and chronic systolic heart failure (CHF) was primarily explained by illness-specific differences related to diagnosis or whether more generic factors also contributed to their physical functioning. Consecutive patients with COPD (n = 56; mean age = 67.8, SD = 8.5) and CHF (n = 65; mean age = 60.0, SD = 10.2)from the outpatient clinics of a university hospital and a general hospital completed a self-report questionnaire, including the Rand-36 Health Survey, Cantril's ladder, the Mastery scale, the Perceived Health Competence Scale, and the Self-efficacy scale. COPD patients scored significantly worse in self-reported physical and psychological functioning and perceived health competence than did patients with CHF Regression analysis revealed that both the diagnosis and the illness severity contributed to self-reported physical functioning, although self-efficacy explained the main part of physical functioning. Therefore, important aims in the treatment of patients with COPD and CHF should be not only improving physical functioning but also enhancing self-efficacy. PMID- 16252623 TI - Mothers' estimates of their children with disorders of language development. AB - The authors' objective in this article was to explore the accuracy of mothers' estimates concerning their children's developmental functioning, especially with respect to vocabulary and gross motor development, by comparing the results of diagnostic tests administered to both the children and their mothers. The authors studied 55 children with disorders of language development (LD) between the ages of 3 and 6 years using several diagnostic scales concerning child development. The authors assessed the mothers' estimates regarding vocabulary and gross motor development by the same scales as those used for their children. These scales were presented as questionnaires and scored like the tests used for the children. There was a significant tendency toward a general overestimation of a child's developmental functioning regarding vocabulary and gross motor skills. Moreover, the accuracy of the mothers' estimates did not seem to be associated with several selected variables. The results did not correspond to those of other studies that have shown that mothers' estimates provide a good indication for the developmental status of a child--at least for the child's developmental status considering vocabulary and gross motor skills in children with LD. The results support the objection regarding the use of maternal estimates as the only source of information concerning the development of a child among scientific studies, especially if they deal with research on the development of vocabulary or gross motor skills. PMID- 16252624 TI - Certification: a credential of professional commitment, competence, and expertise. PMID- 16252625 TI - A review of thyroid hormone administration during adult donor care. AB - Serum concentrations of thyroid hormones tetraiodothyronine and triiodothyronine commonly are low after head injury and brain death. Thyroid hormone replacement therapy, however, is a controversial part of donor management. This article reviews publications in which thyroid hormone administration was evaluated in human donors. A classification of the "quality" of study methods used in those publications is presented as part of the data review. No publications support the routine administration of thyroid hormone for all donors. "Rescue" replacement in support of cardiac inotropic function is supported by some studies, but the experimental design of those investigations is not optimal. Thyroid hormone replacement and its dosing should be decided by organ procurement organizations as part of treatment protocols. PMID- 16252626 TI - Leading the flock: organ donation feelings, beliefs, and intentions among African American clergy and community residents. AB - CONTEXT: Despite a considerable potential role in organ donation for African American clergy, there has been little investigation to date of the beliefs, attitudes, and personal intentions of such clergy regarding donation. OBJECTIVE: To compare the beliefs, attitudes, and behavioral intentions regarding organ donation among African American clergy to those of African American residents of the same large US city. DESIGN: Focus groups and 3 cross-sectional surveys. SETTING: Greater Houston, Tex, metropolitan area. PARTICIPANTS: A total of 761 randomly selected African American community residents and 311 African American clergy. MAIN OUTCOMES MEASURES: Beliefs about the importance of organ donation; how comfortable one is in thinking about donation; whether one believes that organ donation is against one's religion; trust in healthcare professionals regarding death declaration; concerns that donation leads to body mutilation; and the likelihood that one will donate one's own organs upon death. RESULTS: Compared to general African American residents, African American clergy in the Houston area were found more often to believe in the importance of donation; to be more comfortable with thinking about donation; to feel more certain that donation was not against their religion; to believe that they could trust healthcare professionals regarding death declaration; to feel less often that donation leads to mutilation of the body; and to indicate a greater likelihood of donating their own organs upon death. The same was found to be true among clergy and congregants of the largest religious denomination in Houston, the Baptists. PMID- 16252627 TI - Variables during care of adult donors that can influence outcomes of kidney transplantation. AB - Publications that relate characteristics of donors to renal function of recipients are reviewed. Most publications report retrospective observations that relate outcomes to donor variables that cannot be altered during donor care. Factors that can be altered in adult donors in an effort to improve recipients' outcomes include urine output and creatinine level. Increasing urine output to more than 100 mL/h, at least during the hour before explantation, and returning the creatinine level to match its serum concentration when the patient was admitted can improve outcomes. Ways of accomplishing those goals during donor care are discussed, with emphasis on support of renal blood flow. PMID- 16252628 TI - Treatment goals during care of adult donors that can influence outcomes of heart transplantation. AB - Myocardial dysfunction during care of adult donors can result from injury occurring before hospital admission or during the progression of brain death. Few evidence-based data correlate specific hemodynamic goals during donor care with outcomes of heart transplantation, although many recommendations exist. Spontaneous reversal of early heart damage or correction of poor cardiac performance can yield outcomes equivalent to outcomes in recipients who had ideal donors. Hemodynamic goals developed in the operating room can be applied in intensive care to improve outcomes of transplantation. These goals include maintenance of mean arterial pressure greater than 60 mm Hg, central venous pressure less than 12 mm Hg, cardiac output greater than 3.8 L/min, cardiac index greater than 2.1, and systemic vascular resistance between 800 and 1200 dyne x sec x cm(-5). The ejection fraction and other echocardiographic data also provide helpful guidance when determining whether a heart is suitable for transplantation and during therapy. Titration of cardiovascular variables often requires invasive monitoring to ensure that cardiac preload, afterload, and contractility are optimal. PMID- 16252629 TI - Indo-Canadian beliefs regarding organ donation. AB - CONTEXT: There is concern that a disproportionately high number of people of South Asian origin await transplantation in Canada. The donation rate is low in this population, and it is difficult to obtain good tissue matches. OBJECTIVE: To explore the values and beliefs regarding organ donation among Indo-Canadian people living in British Columbia. DESIGN: A naturalistic qualitative study was designed. Individual interviews and focus groups were held to collect data pertaining to beliefs regarding organ donation. SETTING: Lower Mainland of British Columbia, Canada. PARTICIPANTS: A total of 40 Indo-Canadian persons participated; a wide range of ages, religions, and backgrounds were represented. RESULTS: The major themes that emerged from the data related to context (including family/community, religion, trust in the healthcare system, and knowledge about organ donation); and values and beliefs (including intergenerational considerations, death and dying, and the organ donation process). Participants noted that people from their community were reluctant to discuss death and relatedissues such as organ donation. Although there was recognition of the importance of individual decision making pertaining to organ donation, the participants believed that family and community members also should be involved. CONCLUSIONS: Beliefs varied considerably among participants, and one should not make assumptions about the beliefs of any one individual based primarily on that individual's membership in an ethnocultural community. PMID- 16252630 TI - Introduction to the interpretation of chest radiographs during donor care. AB - Organ procurement coordinators often provide independent interpretations of chest radiographs during donor care. Catheter or tube position, lobar atelectasis, extra-alveolar air, air bronchograms, pleural fluid, and other findings are important throughout donor care and when deciding if a lung is acceptable for transplantation. Technical factors, features of a normal chest radiograph, and abnormal radiographic findings are reviewed and examples are presented. PMID- 16252632 TI - Minimally invasive surgery for live kidney donors: techniques and challenges. AB - Live kidney donation is assuming an increasingly prominent role in kidney transplantation programs. The traditional operative approach has been through an incision in the upper quadrant of the abdomen or in the loin, with the attendant potential postoperative complications associated with a large surgical wound. These problems may act as disincentives to prospective donors. The introduction of laparoscopic donor surgery in 1995 heralded a new era offering reduced post operative pain and improved cosmetic result. It is hoped that these benefits may counter some disincentives and thereby increase donation rates. Three minimal access approaches and their advantages and disadvantages are described: classical laparoscopic, hand-assisted laparoscopic, and retroperitoneoscopic surgery. Published reports indicate extensive experience with the first 2 of these approaches and less experience with the latter. All 3 approaches present technical, physiological, and anatomical challenges in the context of retrieving an organ that is fit for transplantation. For minimal-access surgery to be accepted as the procedure of choice for live kidney donors, it must be demonstrated that morbidity is not transferred from donor to recipient when these techniques are used. Some concerns about these procedures are addressed. High level evidence in the form of randomized controlled trials is generally lacking, but experiences of surgeons and patients suggest that, with appropriate modifications, these techniques are safe for both donors and allografts and also benefit donors' recovery. PMID- 16252631 TI - Role of "relatedness" in donation discussions with next of kin: an empirical study of the common wisdom. AB - CONTEXT: Demographic similarities between support personnel and next of kin are of increasing interest. Studies examining like-race requestors have not produced clear and consistent outcomes. No studies have examined demographic relatedness factors for family support personnel who are not requestors. OBJECTIVES: To examine the degree of "relatedness" between family communication coordinator chaplains and next of kin in cases that resulted in consent for donation. To examine "relatedness" in terms of demographic characteristics between primary hospital family support individuals and next of kin. DESIGN: Retrospective nonexperimental descriptive design spanning 1997 to 2004. MAIN OUTCOME MEASURES: Demographic characteristics including gender, race and ethnicity, age, religion, and contact time. RESULTS: The results fail to support the hypothesis that the degree of "relatedness" between the family communicator and next of kin is associated with positive consent decisions. The findings show that gender is a shared quality in 60% of these cases, and race, age, and religion are even more infrequently shared qualities. The findings also suggest an elevated role of women in the donation discussion. CONCLUSIONS: This study fills the void of empirical research through its use of objective outcome measures. The results fail to support the common wisdom and bring to question the foundation of many programs used to promote organ donation. Additionally, the findings emphasize the need for more rigorous investigations and development of more refined, valid measures for examining factors that may influence the donation discussion. PMID- 16252633 TI - Ventricular assist devices: preparing for catastrophic environmental events. AB - In the summer of 2004, Florida experienced 4 major hurricanes in a matter of weeks. These hurricanes left many Floridians without power and passable roads, interrupted communications, and destroyed some homes. During this time, Tampa General Hospital had 1 patient living at home with an implanted ventricular assist device. The patient had been discharged home only 2 weeks before hurricane Frances hit hard. Although the patient was able to stay at home and experienced no major problems with the device, there were several situations that taught us many lessons about caring for patients with ventricular assist devices during environmental catastrophic events. PMID- 16252634 TI - Hand-assisted laparoscopic donor nephrectomy: a low rate of complications. AB - BACKGROUND: Laparoscopic donor nephrectomy is associated with a higher incidence of ureteral complications. Hand-assisted dissection minimizes the use of instruments for intraoperative retraction and handling of periureteric tissue, and may reduce posttransplant complications. OBJECTIVE: To assess the outcome of hand-assisted laparoscopic donor nephrectomy, in particular ureteral complications. METHODS: Records of 143 kidney transplant recipients who received allografts removed using the hand-assisted laparoscopic technique were retrospectively studied. RESULTS: Total operating time was 2.0 +/- 0.55 (range 1.08-4) hours. Warm ischemia time was 1.45 +/- 0.60 (range 0.58-3.00) minutes. Length of artery, vein, and ureter was 2.4 +/- 0.5 cm, 3.0 +/- 0.5 cm, and 10.3 +/- 2.1 cm, respectively. Estimated blood loss averaged 86.3 +/- 55.6 mL. Intraoperative suction was not needed in 65% of patients. Two donors developed incisional hernias and 1 had a postoperative ileus. Four of 143 (2.8%) recipients developed ureteral complications: reoperations for ureteral necrosis (1), stenting for ureteral stenosis (2), and urethral catheterization for ureterovesical leak (1). Graft loss in the first year after transplantation occurred because of renal vein thrombosis, thrombosis of revised arterial anastomosis, arterial thrombosis due to myocardial infarction, vasculitis, focal segmental glomerulosclerosis, and chronic rejection. Delayed graft function developed in 3 recipients. The acute rejection rate was 14.6%. Mean serum creatinine levels at 1 and 3 years were 134 +/- 61 micromol/L (1.52 +/- 0.69 mg/dL) and 121 +/- 35 micromol/L (1.37 +/- 0.40 mg/dL), respectively. CONCLUSIONS: Hand-assisted laparoscopic donor nephrectomy is associated with a low incidence of ureteral complications; may reduce the technical difficulty of the operation and minimize retraction with instruments, resulting in fewer complications for donors and recipients; and minimizes donor blood loss. PMID- 16252636 TI - Coordinating organ transplantation in Turkey: effects of the National Coordination Center. AB - In January 2001, the National Coordination Center, which brought tissue and organ procurement and transplantation under the Turkish Health Ministry, was established in Turkey. The main aims of this organization are to expand cadaveric donation and increase the number of transplantable organs supplied by cadaveric donors. We compared the proportions of cadaveric organ transplantations that were performed in Turkey before and after the national coordination system was established. Of all the cadaveric transplantations completed to date, 91.6% of kidney and 71.5% of liver procedures were done before implementation of the new system, and 8.4% and 28.5%, respectively, were performed after the system was established. The data show that the frequency of cadaveric donation has increased, as well as the number of cadaveric organ transplantations performed annually. The new national transplantation coordination system is making a good start at increasing cadaveric transplantation in Turkey. This system will hopefully lead to a larger organ pool and shorter waiting lists in future. PMID- 16252635 TI - Preoperative predictors for postoperative problems in heart transplantation: psychiatric and psychosocial considerations. AB - The psychiatric and psychosocial evaluation of the heart transplant candidate can identify particular predictors for postoperative problems. These factors, as identified during the comprehensive evaluation phase, provide an assessment of the candidate in context of the proposed transplantation protocol. Previous issues with compliance, substance abuse, and psychosis are clear indictors of postoperative problems. The prolonged waiting list time provides an additional period to evaluate and provide support to patients having a terminal disease who need a heart transplant, and are undergoing prolonged hospitalization. Following transplantation, the patient is faced with additional challenges of a new self image, multiple concerns, anxiety, and depression. Ultimately, the success of the heart transplantation remains dependent upon the recipient's ability to cope psychologically and comply with the medication regimen. The limited resource of donor hearts and the high emotional and financial cost of heart transplantation lead to an exhaustive effort to select those patients who will benefit from the improved physical health the heart transplant confers. PMID- 16252637 TI - Promoting organ donation and transplantation among South Asians in the United Kingdom: the role of social networks in the South Asian community. AB - The percentage of South Asians on the kidney transplant waiting list in the United Kingdom is 3 times their percentage in the general population. Obviously, organ donation and transplantation among South Asians in the United Kingdom needs improvement. In recent years, ethnically targeted campaigns in the mass media have specifically attempted to attract donors from the South Asian communities. A number of pilot studies have been done to evaluate the effectiveness of these initiatives in providing information about organ donation to South Asians. Results indicate that detailed information related to transplantation was learned mainly by people within the community receiving transplants and was transmitted through various informal community networks rather than through the resources provided by the Department of Health. This article provides an overview of who South Asians are and how these community networks were established. Transplant professionals must devise effective strategies to access these community networks, thereby raising the consciousness of transplantation among South Asians in the United Kingdom. PMID- 16252638 TI - The institutional review board and protecting human subjects: 10 frequently asked questions. AB - It is essential that anyone involved in research involving human subjects be familiar with the purpose and role of institutional review boards. Institutional review boards are designed, first and foremost, to protect human research subjects by overseeing the implementation of federal regulations regarding protection of human subjects. The federal government requires institutional review board approval for any human subject research that receives federal funding, and many scholarly journals require proof of institutional review board approval of the research before publication. In this article, the answers to 10 frequently asked questions about the role of institutional review boards highlight the important contributions made by institutional review boards to the conduct of ethically sound research. The aim is to generate a working knowledge of the institutional review board's function that can be used by every researcher contemplating working with human research subjects. This is the first in a series of 3 articles examining common issues in research ethics. PMID- 16252639 TI - Domino kidney transplantation. AB - Kidney transplantation has become a victim of its own success. Despite measures to increase the number of donors, success to date has been limited. At the Western Infirmary, we used an organ that had been transplanted earlier. The patient who received that organ has since been followed up for more than 2 years, and no issues have arisen regarding functioning of the graft. Although it does not increase the donor pool, we believe that domino kidney transplantation ought to be considered when the situation merits it. PMID- 16252640 TI - Role of the independent donor advocacy team in ethical decision making. AB - Adult living donor liver transplantation has developed as a direct result of the critical shortage of deceased donors. Recent regulations passed by New York State require transplant programs to appoint an Independent Donor Advocacy Team to evaluate, educate, and consent to all potential living liver donors. Ethical issues surround the composition of the team, who appoints them, and the role the team plays in the process. Critics of living liver donation have questioned issues surrounding motivation and the ability of donors to provide true informed consent during a time of family crisis. This article will address issues surrounding the controversies and discuss how using the team can effectively evaluate and educate potential living liver donors and improve practice to ensure safety of living donors. PMID- 16252641 TI - Ethical analysis of living organ donation. AB - In 2003, the first 3-way living kidney donor-swap was performed at Johns Hopkins Hospital in Baltimore, Md. Three new donor protocols includingpaired donation now allow unrelated individuals to serve as donors. Some ethicists have suggested that emotionally unrelated individuals not be permitted to donate because they will not experience the same satisfaction that a family member who is a donor experiences. Others who frame living donation as an autonomous choice do not see emotionally unrelated or even nondirected donation as ethically problematic. This article uses an ethical framework of principlism to examine living donation. Principles salient to living donation include autonomy, beneficence, and nonmaleficence. The following criteria are used to evaluate autonomous decision making by living donors, including choices made (1) with understanding, (2) without influence that controls and determines their action, and (3) with intentionality. Empirical work in these areas is encouraged to inform the ethical analysis of the new living donor protocols. PMID- 16252642 TI - Method for embedding optical fibers in an aluminum matrix by ultrasonic consolidation. AB - The overall aim of the research, part of which is outlined in this paper, was to utilize the ultrasonic consolidation (UC) process for the fabrication of smart metal structures, capable of measuring an external stimulus and responding to this stimulus by adapting its structure accordingly through embedding both active and passive functional elements. This paper presents a fundamental study of embedding methods for the fabrication of optical fibers embedded within aluminum structures. The methods considered in this paper produced embedded optical fiber specimens in which large amounts of plastic flow were observed within the matrix. The matrix material deformed around the fibers, resulting in fully embedded optical fibers capable of transmitting a bright light source and without damaging the fibers. Based on light responses, a general process window was drawn to show the range at which optical fibers can be embedded within aluminum structures using the UC process. The outcomes lay down initial investigative principles for the further development of the technology for embedding or cladding of optical fiber sensors, such as fiber Bragg grating devices, within or on metal structures: for example, the cladding of large free-form metal structures or smart "skinned" metal foam or metal honeycomb structures. PMID- 16252643 TI - Sensing of corrosion on aluminum surfaces by use of metallic optical fiber. AB - We present a new method for monitoring aluminum corrosion by determining the kind of light output that is as corrosion occurs. We prepared some metallized multimode optical fibers by physical vacuum deposition of aluminum to monitor metal corrosion. The sensing area was 1-2 cm in length and had an uncladded part. We used scanning-electron microscopy (SEM) to observe the microappearance of the aluminum before and after corrosion by sodium hydroxide or hydrochloric acid. The film's thickness was also measured by SEM. The factors that affect the rate of corrosion were also investigated. PMID- 16252644 TI - Investigation of laser percussion hole drilling by use of speckle correlation. AB - In this paper speckle correlation is introduced as a tool to investigate the heat influenced area during material processing with laser light. Two materials were investigated, a pure silver sheet and a sheet of SiC-diamond composite. The processing laser used in the experiments was a diode-pumped acousto-optical Q switched Nd:YAG laser that allowed percussion hole drilling to be performed using green light through a second-harmonic crystal. The measurements were performed using a continuous-wave He-Ne laser and a digital camera. The experimental results show that the heat-influenced area is approximately 5000 times larger than the actual hole being drilled and that it reaches a steady-state condition toward the end of the processing cycle. PMID- 16252645 TI - Neural network-based image reconstruction for positron emission tomography. AB - Positron emission tomography (PET) is one of the key molecular imaging modalities in medicine and biology. Penalized iterative image reconstruction algorithms frequently used in PET are based on maximum-likelihood (ML) and maximum a posterior (MAP) estimation techniques. The ML algorithm produces noisy artifacts whereas the MAP algorithm eliminates noisy artifacts by utilizing availableprior information in the reconstruction process. The MAP-based algorithms fail to determine the density class in the reconstructed image and hence penalize the pixels irrespective of the density class and irrespective of the strength of interaction between the nearest neighbors. A Hebbian neural learning scheme is proposed to model the nature of interpixel interaction to reconstruct artifact free edge preserving reconstruction. A key motivation of the proposed approach is to avoid oversmoothing across edges that is often the case with MAP algorithms. It is assumed that local correlation plays a significant role in PET image reconstruction, and proper modeling of correlation weight (which defines the strength of interpixel interaction) is essential to generate artifact-free reconstruction. The Hebbian learning-based approach modifies the interaction weight by adding a small correction that is proportional to the product of the input signal (neighborhood pixels) and output signal. Quantitative analysis shows that the Hebbian learning-based adaptive weight adjustment approach is capable of producing better reconstructed images compared with those reconstructed by conventional ML and MAP-based algorithms in PET image reconstruction. PMID- 16252646 TI - Design and fabrication of narrow-frequency sharp angular filters. AB - The theoretical deduction for the frequency variations of the defect mode and its dependence on the incident angle in one-dimensional defective photonic crystals is discussed. Filters with the dual function of narrow frequency passbands and sharp angular pass breadths in visible and near-infrared wave bands, in which both the defect mode and the periodic structure are layers of integral times of quarter-wavelengths, were designed. This kind of filter was fabricated with commonly used coating machines. PMID- 16252647 TI - Thermal cycling and the optical and electrical characterization of self-assembled multilayer nile blue A-gold thin films. AB - Some laser applications produce high power densities that can be dangerous to equipment and operators. We have fabricated thin-film coatings by using molecular electrostatic self-assembly to create a spectrally selective absorbing coating that is able to withstand thermal fluctuations from -20 degrees C to 120 degrees C. We made the thin-film coatings by alternating deposition of an organic dye and gold colloidal nanoparticles onto glass substrates. Nile Blue A perchlorate, with a maximum absorbance slightly above 632 nm, was chosen as the organic dye. Strong coupling between the dye molecules and the gold nanoparticles provides a redshift that increases as the film's thickness is increased. The incorporation of the gold colloidal nanoparticles also decreases the resistivity of the film. The resistivity of the film was measured with a four-point probe and found to be approximately 10 omega/cm for the two samples measured. Atomic-force microscopy was used to show that film thickness increased 2.4 nm per bilayer. The optical properties of the film were measured at the end of every 5 thermal cycles from 20 degrees C to 120 degrees C, and negligible degradation was observed after 30 cycles. PMID- 16252648 TI - Analysis of intrachip electrical and optical fanout. AB - We examine the benefits of electrical isolation in intrachip optical signaling. We calculate the delay and energy metrics of an optical interconnect with fanout driving an electrical load. By examining fanout and including load drivers into delay equations, we make a shift from the general trend of looking at optical interconnects as a replacement for long parasitic wires. Our calculations show that optical fanout provides a large improvement in an Etau2 (energy delay squared) metric and improves performance even at very short intrachip distances. The break-even length corresponds to the wiring length of 250 minimum-size inverters that are compactly laid out. These results provide a compelling reason to further examine the implementation of optical interconnects. PMID- 16252649 TI - Relationship between parameters of bacteriorhodopsin film and behavior of optical novelty filters. AB - To continue our earlier research on novelty filters in a system of incoherent light [Opt. Lett. 30,81 (2005)], we discuss the relationship between parameters of a bacteriorhodopsin film and the quality of a novelty filter image. For both fixed and moving velocities of the input image, differences in the novelty filter's image as a function of thickness, lifetime of the M state, and molecular concentration are displayed, and the optimal ranges of parameters of the bR film that correspond to the entire novelty filter image and obvious gray-level differences in the image are given. The method can be used to design high-quality novelty filter images in incoherent light systems. PMID- 16252650 TI - Analysis of optical channel cross talk for free-space optical interconnects in the presence of higher-order transverse modes. AB - We analyze the effect of cross-talk noise on the performance of free-space optical interconnects (FSOIs). In addition to diffraction-caused cross talk, we consider the effect of stray-light cross-talk noise, an issue that, to the best of our knowledge, was not addressed previously. Simulations were performed on a microlens-based FSOI system using the modal composition and beam profiles experimentally extracted from a commercial vertical-cavity surface-emitting laser. We demonstrate that this cross-talk noise introduces significant degradation to interconnect performance, particularly for multitransverse-mode laser sources. A simple behavioral model is also developed that accurately approximates the cross talk noise for a range of optical sources and interconnect configurations. PMID- 16252651 TI - Atmospheric compensation with a speckle beacon in strong scintillation conditions: directed energy and laser communication applications. AB - Wavefront control experiments in strong scintillation conditions (scintillation index, approximately equal to 1) over a 2.33 km, near-horizontal, atmospheric propagation path are presented. The adaptive-optics system used comprises a tracking and a fast-beam-steering mirror as well as a 132-actuator, microelectromechanical-system, piston-type deformable mirror with a VLSI controller that implements stochastic parallel gradient descent control optimization of a system performance metric. The experiments demonstrate mitigation of atmospheric distortions with a speckle beacon typical for directed energy and free-space laser communication applications. PMID- 16252652 TI - Efficient fabrication of fused-fiber biconical taper structures by a scanned CO2 laser beam technique. AB - The driving mechanism of a scanning mirror can cause significant impairment of expanded beam properties, which we investigated for several scanning waveforms. Engineering on the scanning waveform is then carried out by a scanned CO2 laser beam technique to enlarge the uniform heating region for stretching and sintering of silica fibers. Details of the derivation are given. A simple thermal model is presented to account for the relationship between the scanning beam profile and the taper shape. Fusion profiles are also compared for various scanning waveforms. The corresponding scanned beam power distributions are determined experimentally, which enables us to determine precise power density conditions for CO2 laser fusion. PMID- 16252653 TI - Calibration of a Shack-Hartmann sensor for absolute measurements of wavefronts. AB - We demonstrate a method with which to calibrate a Shack-Hartmann sensor for absolute wavefront measurement of collimated laser beams. Nearly perfect spherical wavefronts originating from a single-mode fiber were used as references. After the calibration, the uncertainty of the wavefront was less than lambda/100 peak to valley across a diameter of 6 mm. For example, this method allowed us to balance aberrations and prepare collimated beams with wavefronts that are plane to lambda/500 across 1 mm. PMID- 16252654 TI - Radiometric validation of NASA's Ames Research Center's Sensor Calibration Laboratory. AB - The National Aeronautics and Space Administration's (NASA's) Ames Research Center's Airborne Sensor Facility (ASF) is responsible for the calibration of several airborne Earth-viewing sensor systems in support of NASA Earth Observing System (EOS) investigations. The primary artifact used to calibrate these sensors in the reflective solar region from 400 to 2500 nm is a lamp-illuminated integrating sphere source. In September 1999, a measurement comparison was made at the Ames ASF Sensor Calibration Facility to validate the radiometric scale, establish the uncertainties assigned to the radiance of this source, and examine its day-to-day repeatability. The comparison was one of a series of validation activities overseen by the EOS Calibration Program to ensure the radiometric calibration accuracy of sensors used in long-term, global, remote-sensing studies. Results of the comparison, including an evaluation of the Ames Sensor Calibration Laboratory (SCL) measurement procedures and assigned radiometric uncertainties, provide a validation of their radiometric scale at the time of the comparison. Additionally, the maintenance of the radiance scale was evaluated by use of independent, long-term, multiyear radiance validation measurements of the Ames sphere source. This series of measurements provided an independent assessment of the radiance values assigned to integrating sphere sources by the Ames SCF. Together, the measurements validate the SCF radiometric scale and assigned uncertainties over the time period from September 1999 through July 2003. PMID- 16252655 TI - Generation of continuously wavelength-tunable optical short pulses by use of two self-seeded Fabry-Perot laser diodes and an optical switch. AB - Generation of wavelength-tunable optical short pulses by use of two self-seeded Fabry-Perot laser diodes in a parallel configuration is described. The system supports continuous wavelength tuning in a relatively wide range of 42 nm. The side-mode suppression ratio achieved is 35 dB across the entire wavelength tuning range. The system is convenient for continuous wavelength tuning. PMID- 16252656 TI - Infrared transmissive, hollow plastic waveguides with inner Ag-Agl coatings. AB - Hollow polycarbonate waveguides with thin-film coatings of Ag-AgI were fabricated by liquid-phase chemistry methods. These hollow waveguides, which have bore sizes ranging from 500 to 2000 microm and lengths as long as 2 m, are transmissive from 2 to more than 20 microm. The lowest loss of 0.02 dB/m was obtained for a straight 2000 microm bore guide at 10.6 microm. This is to our knowledge the lowest loss measured for any IR fiber at CO2 laser wavelengths. The bending losses were found to increase as 1/R, where R is the radius of the bend. These waveguides were able to withstand 18 W of CO2 laser input power for bore sizes greater than 1000 microm. PMID- 16252657 TI - Theoretical models and analytic expressions for buildup time of pulsed confocal unstable optical parametric oscillators with uniform or Gaussian reflectivity mirrors. AB - A theoretical model and a simplified analytic expression are developed to describe the buildup time of a pulsed confocal unstable optical parametric oscillator (OPO) with a uniform-reflectivity mirror (URM) or a Gaussian reflectivity mirror (GRM). Two analytic expressions have been demonstrated to correspond to theoretical models with a sufficient degree of accuracy. The effects of a variety of cavity and pump parameters on the buildup time of an OPO were investigated and analyzed. It was found that a GRM unstable OPO generally exhibits a shorter buildup time than the corresponding URM unstable OPO with equally effective output coupling. PMID- 16252658 TI - Extension of the graphical technique for estimation of particle size distribution parameters for the consistent intercomparison of diverse sets of multiwavelength lidar derived optical coefficients. AB - In applying the graphical technique to the estimation of the particle size distribution (PSD) parameters, determination of proper bounds surrounding the solution space for a particular confidence level is essential to the consistent intercomparison of diverse multiwavelength lidar optical data sets. The graphical technique utilizes ratios of backscatter and/or extinction coefficients, and it is shown that if the correlation between ratios is not taken into account in calculating the error bounds, the solution space will be overestimated, resulting in relatively larger discrepancies for a larger number of optical coefficients. A method for correcting the bounds, to account for the correlation is developed for various numbers of wavelengths. These improved bounds are then applied, for the case of a monomodal lognormal PSD, with an assumed refractive index, to assess the role additional Raman extinction channels play in improving retrieval capability of a typical three-channel backscatter lidar (1064, 532, and 355 nm) under varying noise levels. Applying the same formalism to underlying bimodal distributions of coarse and fine particles can result in false monomodal solutions. However, when both Raman optical extinction channels are available, no solution is obtained. This can potentially serve as a quick and simple method, prior to a more complex regularization analysis, to differentiate between cases in which the fine mode is dominant versus the cases in which the contribution from the coarse mode is significant. PMID- 16252659 TI - Ultraviolet Rayleigh-Mie lidar by use of a multicavity Fabry-Perot filter for accurate temperature profiling of the troposphere. AB - A UV Rayleigh-Mie scattering lidar system at 355 nm has been upgraded for more accurate temperature profiling of the troposphere by use of a new multicavity Fabry-Perot etalon (MCFPE) filter. The MCFPE filter, which was designed to improve the stability and operational characteristics of the lidar system, has three filter bandpass functions and separates one Mie scattering and two Rayleigh scattering signals from the lidar return signal and simultaneously acts as a laser frequency discriminator to lock the laser frequency. Moreover, a high resolution grating is employed to block signal interference from Raman scattering and the solar background. A practical lidar system, which features strong system stabilization and high measurement accuracy, has been built, and the performance of the lidar system has been verified by comparison of temperature profiling between the lidar and a radiosonde. Good agreement between the two instrument measurements was obtained in terms of lapse rate and inversion layer height. Statistical temperature errors of less than 1 K up to a height of 3 km are obtainable with 5 min observation time for daytime measurements. PMID- 16252660 TI - Discrete dipole approximation simulations of scattering by particles with hierarchical structure. AB - We use the discrete dipole approximation (DDA) method to calculate the intensity and the linear polarization degree of light scattered by agglomerated debris particles with hierarchical structure as functions of size parameter (varying from x = 2 to x = 14) and phase angle. Such structures are important, e.g., for cometary and interplanetary dust particles. Calculations for three combinations of refractive index were made, which correspond to regions of water ice, organic matter, and silicates. We examine the photometric and polarization properties of agglomerated particles with prefractal (Whitten-Sander model) and nonfractal porous structures of particle fragments formed by dipoles. We find that the aggregated particles can produce significant negative polarization at small phase angles. Increasing the packing density of dipoles and/or refractive index makes the negative polarization more prominent. The depth of the negative polarization branch depends on the type of internal structure: the negative polarization branch of particles having nonfractal structure is noticeably shallower in comparison with that of those having a prefractal structure. The negative polarization branch depth strongly depends on the imaginary part of the refractive index and increases with decreasing absorption. Polarization phase curves for agglomerated debris particles become smoother as the number of hierarchical levels increases. PMID- 16252661 TI - Principal-components analysis of fluorescence cross-section spectra from pathogenic and simulant bacteria. AB - Principal-components analysis of a new set of highly resolved (< 1 nm) fluorescence cross-section spectra excited at 354.7 nm over the 370-646 nm band has been used to demonstrate the potential ability of UV standoff lidars to discriminate among particular biological warfare agents and simulants over short ranges. The remapped spectra produced by this technique from Bacillus globigii (Bg) and Bacillus anthracis (Ba) spores were sufficiently different to allow them to be cleanly separated, and the Ba spectra obtained from Sterne and Ames strain spores were distinguishable. These patterns persisted as the spectral resolution was subsequently degraded in processing from approximately 1 to 34 nm. This is to the author's knowledge the first time that resolved fluorescence spectra from biological warfare agents have been speciated or shown to be distinguishably different from those normally used surrogates by optical spectroscopy. PMID- 16252662 TI - Detection of sputtered metals with cavity ring-down spectroscopy. AB - We report on use of cavity ring-down spectroscopy (CRDS) as a means to detect and quantify ion sputtering of refractory metal species. CRDS measurements are made with a neodymium:YAG-pumped optical parametric oscillator laser system in the 375 400 nm region. CRDS sputtering measurements are presented for argon ions incident on iron, aluminum, molybdenum, and titanium. The measurements are based on absorption from fine-structure levels of the electronic ground-state multiplets. For each species, characteristic spectra are provided, the dependence of sputtered particle number density on the beam current is examined, measured densities are compared with a sputter model, and detection limits are determined. For iron, aluminum, and titanium we probe multiple fine-structure levels within the ground-state multiplet and obtain information on their relative populations. PMID- 16252663 TI - Femtosecond laser speckles. AB - The concept of femtosecond laser speckles is put forward. The theory of a speckle pattern in light of finite bandwidth is applied to describe femtosecond laser speckles. Basic representations of the contrast and the spectral correlation of femtosecond laser speckles are presented. The relationship between the speckle contrast and the bandwidth of a femtosecond laser is given. Experimental results are given that indicate an obvious difference between the speckle patterns produced by a continuous-wave laser and those produced by a femtosecond laser. PMID- 16252664 TI - Synthesis of novel alpha-L-arabinopyranosides of beta-lactams with potential antimicrobial activity. AB - Synthetic routes toward the synthesis of some novel 1-(2,3,4-tri-O-acetyl-alpha-L arabinopyranosyl)-azetidin-2-ones are described. Antimicrobial screening of three selected compounds revealed their activity against Bacillus subtilis and Escherichia coli PMID- 16252665 TI - Unusual olefin formation by PhSe-F trans-elimination. AB - A new approach to the synthesis of 2',3'-didehydro-2',3-dideoxynucleosides was described in excellent yield through unusual olefin formation by PhSe-F trans elimination. PMID- 16252666 TI - Polysulfide reagent in solid-phase synthesis of phosphorothioate oligonucleotides: greater than 99.8% sulfurization efficiency. AB - A solution of sulfur (0.1 M) and sodium sulfide (0.01M) in 3-picoline, referred to as polysulfide reagent, rapidly converts trialkyl and triaryl phosphite triesters to the corresponding phosphorothioate derivatives. Greater than 99.8% average stepwise sulfurization efficiency is obtained in the solid-phase synthesis of DNA and RNA phosphorothioate olgonucleotides via the phosphoramidite approach. PMID- 16252667 TI - A proposal for a new stereochemical notation for P-chiral nucleotide analogues and related compounds. AB - A new stereochemical notation for P-chiral nucleotide analogues and related compounds is proposed In this notation, the names of configurations, designated as D(P) and L(P), are derived from a geometrical relationship, rather than from priority rules, of substituents at the phosphorus centre. This new stereochemical description offers clear advantages over the CIP R/S nomenclature, particularly when used for comparing the influence of absolute configuration at the phosphorus centre on physicochemical and biological properties of oligonucleotide analogues or in stereochemical correlation analysis of P-chiral nucleotide derivatives. PMID- 16252668 TI - Synthesis and evaluation of RNA transesterification efficiency using stereospecific serinol-terpyridine conjugates. AB - Six novel artificial ribonucleases were synthesized employing a stereochemically pure abasic serinol backbone residue for attachment of the RNA transesterification agent copper(II) terpyridine. These stereochemically pure abasic residues were synthesized as phosphoramidite building blocks from the parent L-serine and D-serine starting building blocks and incorporated into oligonucleotides via solid-phase DNA synthesis. These artificial ribonucleases were constructed to determine if the stereochemistry of the alpha carbon of an abasic serinol residue has influence over RNA transesterification through selective placement of a pendant transesterification agent in either the major or minor groove. The novel artificial ribonucleases and previously synthesized artificial ribonucleases were challenged with a 28-mer and 159-mer RNA substrate. It was determined that the stereochemistry of the carbon atom derived from the alpha-carbon of serine did not influence the extent of cleavage in these studies using copper(II) terpyridine conjugated artificial ribonucleases. PMID- 16252669 TI - New and one pot chemoselective synthesis of nucleoside 5'-H-phosphonate diesters. AB - Arbuzov reaction of phenyl phosphorodichloridite with two equiv of alcohol, or mixture of one equiv of alcohol and one equiv of tert-butyl alcohol led to the corresponding aryl H-phosphonate diesters. Following displacement of the H phosphonate diesters with unprotected nucleosides chemoselectively produced nucleoside 5-H-phosphonate diesters in good yields, respectively. PMID- 16252670 TI - 4-aminometyl-3-nitrobenzoic acid--a photocleavable linker for oligonucleotides containing combinatorial libraries. AB - We detail the design, synthesis, and characterization of an o-nitrobenzyl-based photolabile linker containing amine and carboxyl anchor groups. A model nucleoside monomer modified with an imidazole residue and a precursor unit linked to a heterocyclic base through a photolabile tether is constructed Upon UV irradiation (313- 365 nm), the imidazole containing part of this molecule is released. PMID- 16252671 TI - One-pot synthesis of TBMPS (bis [tert-butyl)-1 pyrenylmethyl-silyl) chloride as a novel fluorescent silicon-based protecting group for protection of 5'-OH nucleosides and its use as purification handle in oligonucleotide synthesis. AB - An efficient and novel synthesis of bis(tert-butyl)- 1-pyrenylmethyl-silyl group (TBMPS) has been reported having fluorescent properties conferred by the pyrenyl group. This silyl group being base labile is efficiently used for one-pot protection of the 5-OH of the nucleosides. While incorporated terminally at the 5 OH of long sequences viz. AA TGG AGC CAG T and GC TAT GTCAGT TCC CCT TGG TTC TC, this group is also helpful in subsequent purification by HPLC as well as PAGE. Besides these, a labeled dimer (T*T) and a labeled tetramer (T*TTT) were also synthesized to compare the fluorescence properties of short and long labeled sequences. Fluorescence properties of these sequences were studied in detail to find the suitability of the approach. PMID- 16252672 TI - Synthesis of some new 1,3/ or 1,4-bis(glucopyranosyl-1,2,4-triazol-5 ylthio)propanes/ or butanes as potential antimicrobial agents. AB - Glucosidation of the appropriate 1,3 or 1,4-bis(4-amino or arylideneamino-2,4 dihydro-3-thioxo-3H-1,2,4-triazol-5-ylthio)propanes or butanes with 2,3,4,6-tetra O-acetyl-alpha-D-glucopyranosyl bromide followed by chromatographic separation gave the corresponding N-, S-, and N,S-bis(glucosides). Chemical transformation leading to new functionalities has been achieved. Antimicrobial screening of 10 selected compounds resulted in their activity against Aspergillus fumigatus, Penicillium italicum, Syncephalastrum racemosum, Candida albicans, Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus subtilis, and Escherichia coli. PMID- 16252673 TI - Chemistry and dynamics of interaction of nucleosides with pentafluoropyridine. AB - Interaction of pentafluoropyridine with hydroxyl groups of thymidine, uridine, adenosine, and deoxyadenosine at room temperature leads to the formation of aryl ethers of nucleosides with a high yield. Under severe conditions, one more tetrafluoropyridine residue is attached to pyrimidine fragments of T and U, while purine heterocycle in A remains intact. Nucleoside derivatives are formed with a quantitative yield and can be used in situ as intermediates for, as an example, molecular design of arene analogs of nucleic acids. The reaction with thymidine is a successive-parallel process, the limited stage being arylation of the secondary hydroxyl group. The presence of the vicinal hydroxyl group in pentose results in the opposite rate ratio of the formation of primary and secondary tetrafluoropyridyl ethers of adenine and uridine. PMID- 16252674 TI - The influence of words as determinants of U.S. international and domestic health policy: part I. PMID- 16252675 TI - Testing for time-based correlates of perceived gender discrimination. AB - Using a sample of 201 medical technologists (MTs) over a five-year period, this study extends initial findings on perceived gender discrimination (PGD) by Blau and Tatum (2000) by applying organizational justice variables and internal external locus of control as hypothesized correlates of PGD. Three types of organizational justice were measured: distributive, procedural, and interactional. General relationships found include locus of control being related to PGD such that internals perceived lower PGD. Also, distributive, procedural, and interactional justice were negatively related to PGD. However, increasing the time interval between these correlates weakened their relationships. The relationship of interactional justice to PGD remained the most "resistant" to attenuation over time. PMID- 16252676 TI - Assessment of allied health graduates' preparation to integrate genetic knowledge and skills into clinical practice. AB - Allied health professionals are in a unique position to address the concerns of and provide information to clients with genetic disorders. This study assessed the preparation of recent graduates of allied health training programs to provide these services by determining their (1) professional practices, (2) confidence in performing skills that require genetic knowledge, (3) extent of genetic training, and (4) interest in genetic topics. A survey was sent to 698 alumni of six allied health training programs who graduated between 1997 and spring 2002 from a midwestern university. A total of 235 alumni responded to the survey (34%). Forty three percent of respondents reported discussing at least one of eight topics, such as patterns of inheritance, recurrence risks, genetic testing, and characteristics of genetic conditions and/or prognosis. Referrals for genetic services were made by 12.6% of respondents. Of the 10 genetic assessment, counseling, and referral skills that were assessed, eliciting family history was the skill most commonly performed. Many respondents discussed the genetic basis of disorders, provided guidance to clients about the impact of their condition, and corrected misconceptions. Only 22% of the respondents rated the amount of genetic knowledge/skills covered in training as satisfactory, and 78% rated it as marginal or none. However, there is strong interest in genetic topics, especially related to common disorders. A correlation was found between the respondents' training and confidence in performing these skills. Allied health professionals are providing genetic-related services in clinical settings. However, sufficient instruction in genetic knowledge and skills is not being provided in their undergraduate and graduate training programs. Preservice and continuing educational interventions designed to prepare allied health professionals for the genetic age are needed. This study establishes baseline data regarding professional practices and perceived clinical confidence related to genetics that can be used to develop and evaluate the effectiveness of future educational interventions. PMID- 16252677 TI - Library-sponsored instruction improves core informatics competencies among allied health students: a research-based case study. AB - Since 1999, a librarian-administered course entitled Computer Literacy for Healthcare Professionals has been required of all incoming occupational, physical, and respiratory therapy students at Stony Brook University. Outcome data from pretests/posttests and student self-assessments over a 3-year period from 2000 to 2002 demonstrate the effectiveness of the training program and of library-sponsored instruction. This study evaluated whether curriculum-integrated informatics training by librarians is effective in enabling allied health students to acquire and retain literature-searching skills. Pretests and posttests measured the impact of instructional intervention upon student acquisition of literature-searching skills; a second posttest ("post5-test") measured skill retention. The intervention group consisted of 179 subjects enrolled in baccalaureate occupational, physical, and respiratory therapy programs between 2000 and 2002; the control group consisted of 48 physician assistant students who had no formal instruction in literature searching. Student self-assessments provided qualitative outcome measures. Paired t test analysis showed that the mean posttest score for the intervention group was considerably greater at the p < 0.01 level (t = 14.868; p < 0.001) than the mean pretest score. The change is statistically significant, and the null hypothesis can be rejected. The results of this study support the conclusion that library-sponsored informatics instruction is effective in improving student acquisition and retention of literature-searching skills. PMID- 16252678 TI - Recruitment and retention of emergency medical technicians: a qualitative study. AB - Emergency medical technicians (EMTs) are critical to out-of-hospital care, but maintaining staff can be difficult. The study objective was to identify factors that contribute to recruitment and retention of EMTs and paramedics. Information was drawn from three focus groups of EMT-Basic, EMT-Intermediate, and EMT Paramedic personnel recruited from participants at an annual conference. Thoughts and feelings of EMTs and paramedics were investigated using eight questions designed to explore entry into emergency medical services, what it is like to be an EMT or paramedic, and the EMT educational process. Data were analyzed at the group level for common themes using NVivo. For a majority of respondents, emergency medical services was not a primary career path. Most respondents entered the industry as an alternate or replacement for a nursing career or as a second career following military medic service. The majority of respondents believed the job was stressful yet rewarding, and although it negatively affected their personal lives, the occupation gave them a sense of accomplishment and belonging. Respondents expressed a preference for EMT education resulting in college credit or licensure versus professional certification. Job-related stress produced by numerous factors appears to be a likely contributor to low employee retention. Recruitment and retention efforts should address study findings, incorporating key findings into educational, evaluation, and job enhancement programs. PMID- 16252679 TI - Challenges of preparing allied health professionals for interdisciplinary practice in rural areas. AB - Meeting the health needs of individuals in rural communities involves addressing the challenges of complex multifaceted health problems, limited local health resources and services, isolation, and distance. Interdisciplinary collaboration can create solutions to health care problems that transcend conventional, discipline-specific methods, procedures, and techniques. This paper reports on the four-pronged approach of the Western Maryland Area Health Education Center used to prepare allied health students to be interdisciplinary team members in rural areas. It describes the development of four interdisciplinary instructional team member training venues (in-class instruction, Web-based modules, service learning programs, and faculty development workshops) that integrate opportunities to develop and practice interdisciplinary health promotion skills in rural communities. Challenges to implementing the model are described, including developing faculty and student training participation, integrating training venues into existing programs at participating institutions, and designing a unified program evaluation. PMID- 16252680 TI - Methods to improve teaching interdisciplinary teamwork through computer conferencing. AB - This study evaluated how initiatives that oriented students to the virtual classroom and faculty feedback that promoted increased student interaction affected teaching teamwork skills in a 6-week, interdisciplinary health professions module. Outcomes from a control group were compared with those from an experimental group that had greater technical support, a print-based study guide, enhanced faculty guidance, and weekly grades rewarding student interaction. Two researchers independently recorded the number of messages each student wrote and coded faculty message content. A t test compared the difference in students' pretest and posttest scores on the Interdisciplinary Education Perception Scale (IEPS) between groups. Fisher exact test was used to compare the total student messages, total faculty messages per team, and faculty message content between modules. The experimental group had greater IEPS belief changes and more faculty messages per team but no difference in total student postings. Faculty messages in the experimental group contained fewer vague messages and more messages containing introductions, restated assignments, reinforcement, and technical information. Providing students with resources to aid their orientation to the virtual classroom and coaching faculty to increase student interaction improved learning outcomes. Faculty messages that restated assignments and contained reinforcement, introductions, and technical information impacted student participation the most. PMID- 16252681 TI - Learning styles of allied health students using Kolb's LSI-IIa. AB - The purpose of this study was to assess the learning style preferences of students enrolled in various allied health professional programs. Five professions were examined: nursing, physician assistant (PA), occupational therapy (OT), physical therapy (PT), and speech-language pathology (SLP). The assessment instrument used was the Kolb Learning Style Inventory LSI-IIa. The study included 89 subjects from the various allied health care programs enrolled at a small midwestern university. Findings indicated similar learning style preferences between nursing, OT, PA, and SLP student groups. Students from these groups exhibited a close balance between all four learning styles. The nursing and SLP groups showed a slight preference for concrete experimentation, whereas the OT and PA groups preferred abstract conceptualization. The learning style of the PT students was that of converger, with a strong tendency toward active experimentation versus reflective observation. An emphasis needs to be placed on student learning styles and its impact in the educational process. An understanding and incorporation of learning styles in the education of health care providers could have a positive impact not only on the teaching and learning process but also on the effectiveness of interdisciplinary team interactions and the patient educational process. PMID- 16252683 TI - [Mechanisms of determining target locator velocity by dolphins and technical applications in hydrolocation and radiolocation]. AB - Dolphin's (Tursiops truncatus) capacity to discriminate between dynamic characteristics of the object location (the target moving radially) was studied. The dolphin sensitivity thresholds for target velocity (2.6 cm/s) and for target acceleration (0.6 cm/s2) were measured. It has been shown that the animal emits two-pulse probe signal to locate the target moving with constant velocity, or three-pulse probe signal--to locate the accelerated target. New highly efficient technical methods of hydro- and radiolocation were suggested on the basis of these peculiarities. PMID- 16252682 TI - [Disorders in learning and memory processes in the monkey model of Alzheimer's disease: the role of the cerebral cortex associative areas]. AB - Processes of novelty learning and keeping the results in Alzheimer's disease in two groups of rhesus-monkeys (three monkeys in each group), were studied: following neurotoxins administrati- on (I group) and saline administration (II group). In two months after the injections (the C1 stage), considerable differences between the groups were revealed in the task of differentiation among contour shapes. For the I group monkeys the learning was difficult: the correct decision making did not reach 85 %, and the probability of refusing to make a decision increased. For the II group monkeys the learning characteristics were not disturbed. In six months after the injections (the C3 stage) the differences between the groups in the task of differentiation among new stimuli (heads of two monkeys) remained at the same level. When studying characteristics of the operative memory associated with keeping the learning results achieved at the C1 stage, a considerable worsening of these characteristics was revealed: diminishing of the correct decision making probability at the C1 stage (actually to the level of 0.5), increase in the probability of refusing to make a decision. The structural-functional organization of interaction between sensory and cognitive processes in learning and keeping the information in the operative memory, is discussed in association with the control of motivation and attention system and the role of the cortex associative areas. PMID- 16252684 TI - [Influence of vibration on the spontaneous neuronal activity of the neurons of the superior vestibular nucleus]. AB - The results of vibration action (5, 10 and 15 daily) on spontaneous neuronal activity of superior vestibular nucleus were studied using software for biological signals. Average histograms and autocorrelograms were drawn up after computerized interspike interval analysis. The results have demonstated that neurons of superior vestibular nucleus have mean frequency 14.0 +/- 1.4 Hz. The results obtained suggest significant reconstruction of spontaneous impulse activity in neurons of the superior vestibular nucleus within postvibration period. PMID- 16252686 TI - [Adrenal glucocorticoid function in rats with various ethanol-induced sleeping time following ethanol administration]. AB - The effect of single and chronic ethanol (Eth) administration (25 % solution, 3.5 g/kg) on functional activity of the hypophyseal-adrenal system in rats with different sensitivity to the hypnotic action of ethanol (short-sleep - SS; non sleep--NS, long-sleep--LS, intermediate group--IG), was studied. It has been shown that, after a single Eth administration, the concentration of corticosterone (K) in LS rat plasma was 1.5-fold higher than that in the NS animals although it did not differ from the K level in SS and Ig those. After repeated ethanol load, the corticosterone contents in the NS rat blood plasma was 3.5-fold and 4.9-fold lower compared to the control and LS groups, respectively. The data obtained indicate that the SS and LS animals had initially different basal blood plasma glucocorticoid level. The SS animals showed a decreased blood plasma K, whereas the LS ones--an increased one. The features of the glucocorticoid status are suggested to be a factor determining the sensitivity of rats to the ethanol hypnotic effect. PMID- 16252685 TI - [Mechanisms of inhibition of the contractile activity in the ileo-caecal zone in rabbits under psychogenic stress]. AB - In experiments on unanaesthetized rabbits, myoelectric activity (contractile activity index) of distal ileum, caecum, and proximal colon in two sites was studied under stress induced by fastening a rabbit to the table in supine position. The stress caused sharp decrease (up to complete disappearance) of the contractile activity in all studied compartments of the ileocaecal intestine with partial or complete restoration after release of the animal. Nonselective blockade of pre- and postsynaptic alpha-adrenoceptor with dihydroergotoxin abolished the initial component of the specified inhibitory response. The latter was caused by "adrenergic inhibition" as a result of action of catecholamines circulating in blood on inhibitory smooth muscle alpha-adrenoceptor. Against the background of muscarinic cholinoceptor blockade, the stressor inhibition of ileocaecal contractile activity observed in control experiments was completely preserved. The periods of supression of ileoceacal contractile activity under stress resistant to blockade of alpha-, beta-adrenoceptor and muscarinic cholinoceptor, are caused by the mechanism of "nonadrenergic noncholinergic inhibition", which is realized at the expence of activation of the enteric inhibitory neurones. PMID- 16252687 TI - [Glucocorticosteroid status and free amino acid level in blood plasma of rats under increased tolerance to ethanol]. AB - The data are presented on the effects of ethanol treatment (3.5 g/kg of a 25 % solution, singly, daily, intraperitoneally for 1, 4, 7, 10 days) on development of tolerance to ethanol as assessed by changes in ethanol-induced sleep, corticosterone level dynamics and free aminoacis content in rat blood plasma. PMID- 16252688 TI - [Effects of caspase-3 inhibition on long-term potentiation in the hippocampus: analysis of paired pulse facilitation]. AB - Incubation of the rat hippocampal slices with caspase-3 inhibitor Z-DEVD-FMK resulted in a time-dependent decrease in long-term potentiation (LTP) magnitude. Analysis of paired pulse facilitation at a 70-msec interval revealed that, after caspase-3 inhibition, the increase in the amplitude of the second response in the pair during LTP that was characteristic for control slices, did not occur. In this situation, the LTP magnitude depended on differences in the amplitudes of the first and second responses before the LTP induction. LTP was absent in slices with initially high efficacy of the afferent stimulation and respective low paired pulse facilitation. The Caspase inhibition seems to prevent structural reorganization during the LTP related to involvement into the response of new synapses and neurons. PMID- 16252689 TI - [Natural killers and biogenic amines: the paracrine regulation in the immune system]. AB - Natural killers are referred to the category of cells well known for their assistance in cyto-differentiation control. Mechanisms of such assistance include cytolytic and secretory effects on the target. In this aspect, the NK capacity to assist in local aminoregulatory processes by adrenalin, serotonin, histamine release contained in these cells granules is considered to be the least studied and the most problematic issue. The solution of this problem demands taking into consideration a number of conditions: the NK ability to receipt biogenic amines and peculiarities of receptor structure; principal differences in mechanism of action of biogenic amines which enter the cell through exogenous pathway or which are synthesized endogenously; correlation between biogenic amines presence in the granules and functional NK state. Hypothesis of generation connected with the problem suggests the NK assistance in cytolysis in concomitant mechanisms of paracrine regulation of cellular microenvironmental proliferative response, induction of local inflammatory and immunomodulating processes. PMID- 16252690 TI - [Influence of local administration of apomorphine on citrulline extracellular level in the striatum: participation of the dopamine D1 and D2 receptors]. AB - By means of in vivo microdialysis combined with HPLC analysis, we have shown that local infusions of 1 mM N-nitro-L-arginine (NO-synthase inhibitors) in the rat striatum reduced, and infusions of 100 microM apomorphine (agonists of the dopamine receptors) increased the level of citrulline (a NO co-product) in extracellular space of this structure. The apomorphine-induced increase in citrulline extracellular levels in the striatum was completely prevented by infusions of N-nitro-L-arginine in this structure, and 10 microM raclopride (dopamine D2 receptor blocker), but not by infusions of 50 microM SCH-23390 (dopamine D1 receptor blocker). The data obtained suggest that the increase in citrulline extracellular levels in striatum resulted from local activation of NO synthase, and this effect is mediated by D2 rather than D1 dopamine receptors. PMID- 16252691 TI - [Interhemispheric connections in the visual cortex of cats reared with bilateral strabismus]. AB - We have determined the spatial distribution of retrograde labelled callosal cells after microiontophoretic horseradish peroxidase injections into the single cortical columns of area 17, 18 in cats reared with bilateral convergent strabismus. The obtained strabismus angle was in the range 10-35 degrees. The zone of labelled cells was located asymmetrically in respect to location of injected column in opposite hemisphere. Some cells were revealed in the transition zone 17/18 and their retinotopic coordinates corresponded to the injected column, as was shown in intact cats. Other labelled cells were located in areas 17, 18, in clusters approximately in 1000 mkm from marginal clusters of transition zone. Analysis of labeling in lateral geniculate nucleus has shown that most of the injected columns were driven by ipsilateral eye. The data obtained may be interpreted as evidence of eye-specificity of monosynaptic callosal connections. The functional role in such connections changes in cats with bilateral strabismus is discussed. PMID- 16252692 TI - [Objective measurements of visual acuity by visual evoked potentials]. AB - Electrophysiological measurements of the threshold spatial frequency were conducted in 26 healthy subjects by using visual evoked potentials with the purpose of objective determination of the visual acuity. For that we proposed a universal method of the visual stimulation and EEG processing (using ICA decomposition in particular) to minimize errors arising on account of individual differences in the visual system functioning. As a result, a correlation of 0.74 and a logarithmic dependence were obtained between spatial resolution measured by electrophysiological and psychophysical methods. The proposed methods of objective measurement of visual resolution has a high effectiveness, does not depend on specificity of individual EEG and domination of different channels in the visual system. Therefore it is possible to determinate objectively the visual acuity in humans independently of their responses. PMID- 16252693 TI - [Effect of rapid and slow cooling on thermoregulatory responses in hypertensive and normotensive rats after calcium administration]. AB - The effect of iontophoretic administration of calcium ions to skin in the area of cold stimulus application on the thermal thresholds and the magnitude of cold defense responses in normotensive Wistar and hypertensive ISIAH rats was studied. In thermoneutral conditions, administration of calciumions wos without effect on the measured thermoregulatory parameters. Under the effect of calcium, the thresholds of all the thermoregulatory responses to cooling (such as heat loss, oxygen consumption, shivering) are lowered and the values of heat loss and shivering thermogenesis are considerably increased. The effects of calcium on thermoregulatory responses depend on the rate of cooling. All changes are more expressive in hypertensive rats. The increased sensitivity of hypertensives to calcium suggests that change in their calcium metabolism may be a cause of the observed shifts in the thermoregulatory response to cold. PMID- 16252694 TI - Potential effect of inter-genic action on peak bone mass (PBM) in Chinese females. AB - Peak bone mass (PBM) is a complex trait, determined by both genetic and environmental factors and also their interactions. Vitamin D receptor (VDR) estrogen receptor alpha (ERalpha), interleukin 6 (IL6), parathyroid hormone (PTH), collagen type I alpha 2 (COL1A2), bone Gla protein (BGP), alpha2-HS glycoprotein (AHSG) are among the important candidate genes of bone metabolism. The study aims to detect significant effect of potential inter-genic action underlying PBM in Chinese females. 361 unrelated healthy premenopausal Chinese females (aged 20 -44 years) with Han ethnicity were recruited from the Shanghai city in China. Bone mineral density (BMD) at the hip and the lumbar spine (L1-4) was measured using a Hologic QDR 2000 + dual-energy X-ray absorptiometry (DXA) scanner. Eight polymorphisms among the seven genes were genotyped, i. e. Apa I in VDR, Pvu II and Xba I in ERa (ERX and ERP, respectively), BsrB I in IL6, BstB I in PTH, Msp I in COL1A2, Hind III in BGP, and Sac I in AHSG, using PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) methods. Two way analysis of variance (ANOVA) showed significant effects of IL6 x ERP interaction on PBM at the total hip (P = 0.019), intertrochanter (P = 0.016), and femoral neck (P =0. 019). The BMD difference between GGPp carriers and GGpp subjects (at these two loci) amounted to 18.0%, 19.5%, and 14.8% at the hip,intertrochanter,and femoral neck,respectively. The potential interaction effect of AHSG x IL6 was observed on femoral neck PBM (P = 0.046). GGSS individuals (at these two loci) had, on average, 18.8% higher femoral neck BMD than those subjects with GGSs genotype. The population-level statistical analysis indicates that IL6 x ERP and AHSG x IL6 have significant inter-genic effect on the genetic determination of PBM in Chinese females. PMID- 16252695 TI - A novel mutation of estrogen receptor gene detected in girls with precocious puberty. AB - Female precocious puberty is caused by premature activation of the hypothalamic pituitary-gonadal axis, exposure to exogenous sex steroid hormones, and the presence of endogenous sex steroids caused by various factors. Estrogen is the final key factor to start onset of puberty. However,in some cases of precocious puberty in girls estrogen elevation could not be detected. The raised sensitivity of estrogen receptor, which may caused by ESR1 mutation or polymorphism, has been frequently mentioned for interpreting the etiology of sporadic low estrogen type cases. But no case evidence has been found in clinical practice. For the purpose of screening possible mutations in estrogen receptor gene, leukocyte genomic DNA were collected from 16 girls with precocious puberty of sporadic low estrogen,and exons of ESR1 were amplified and analysized using PCR-SSCP/silver staining method. A single strand conformation change in exon 8 was found in one of the patients (No. 14). The suspected fragment were cloned to a T vector and sequenced for analysis. Sequencing of these clones revealed that this conformation change is caused by a C to T transition. This mutation results in the replacement of arginine by cystine at position 548 of ESR1 protein. The mutation created an extra Btsl digest site and made it can be readily identified by PCR-PFLP method. Further detection using this method, and sequencing of cloned exon8 colonies from patients proved that the patient No. 14 is Arg548/Cys548 heterozagous in genotype. This mutation increased hydrophobility of the area dramatically. The position and the conservative of this residue in vertebrates suggested Arg548 may play an important role in ESR1 function. For study the role of this mutation in the onset of precocious puberty, a firefly luciferase reporter plasmid pGL3 promoter-ERE was constructed,and a pCR3. 1-hermut pisimid expressing Cys548 ER was constructed based on wild type pCR3. 1her. Co-transfection of reporter and pCR3. 1 -hermut in CMF-7 cell strain proved that Cys548 mutant can significantly increase the transcription activity over the Arg548 wild type. PMID- 16252696 TI - Features of coding and noncoding sequences based on 3-tuple distributions. AB - The origin of non-coding sequences, especially introns,is an outstanding issue that has been receiving continuous debate for the last two decades. In the current work we use a mathematical model to characterize DNA sequences and find that the 3-tuple distributions in different reading frames of a given coding sequence differ sharply from each other, while they are almost identical to each other in introns or other non-coding sequences. SREs (Symmetric relative entropies) decrease progressively from coding sequences of primitive prokaryotes to those of advanced eukaryotes and from non-coding sequences of low eukaryotes to those of high eukaryotes with a correlation coefficient of 0.86. In silico evolution experiments show that SREs typical of higher eukaryotic introns can be achieved from prokaryotic coding sequences as the mutation ratio reaches 2/100. The fact that (a total of 25 introns) from all three different genomes S. pombe, C. elegans and H. sapiens searched are found to share high sequence identity with coding regions indicates that at least some introns may have come directly from CDS (coding sequences). We suggest that SREs may be a useful feature for evolutionary study. PMID- 16252697 TI - A simple method to detect the heterogeneity of nucleotide substitution processes by measuring asymmetry in paired comparison. AB - A simple method is presented, which uses a chi2 statistic to measure asymmetry of the substitution matrix between two DNA sequences in order to test a homogeneity hypothesis of the substitution processes. In theory, this chi2 test holds irrespective of whether there is among-site (i) heterogeneity in substitution rates, (ii) correlation in evolutionary rates/models, and (iii) variation in substitution models. Computer simulations showed that the chi2 test is powerful under a variety of models of sequence evolution. Comparison of the eleven sequenced arthropod mtDNAs by using this test revealed that most of the observed evolutionary models were homogeneous between the two mosquitoes but not between Daphnia pulex or Artemia franciscan and the other arthropods, probably due to shifts to a high AT content. A comparison to Kumar and Gadagkar's test by computer simulation as well as empirical data analysis is also given. PMID- 16252698 TI - Microsatellite analysis of royal jelly producing traits of Italian honeybee (Apis mellifera Liguatica). AB - Genetic variations at 10 microsatellite loci were surveyed to determine the evolutionary relationships and molecular characteristics of three different honeybee (Apis mellifera L.) populations from Italy and China, i. e., native Italian (Ee), Chinese-Italian (Eb) and selected high royal jelly producing bees (Ea). A total of 96 alleles,an average of 9.6 alleles per locus,were scored in Ee,Eb and Ea bees at 10 loci. Out of which 48 (5%) were different. This indicated a high degree of polymorphism and ever, some genetic differentiation among the three populations due to artificial selection and geographical isolation. The polymorphic information contents (PIC) and heterozyosity of the three populations at 10 loci were 0.57, 0.50, 0.57, and 0.60, 0.57, 0.61, for Ee, Eb, Ea populations respectively, neither of which were different. This indicated same gene diversity within the three populations. The genetic distance was shorter between Ee and Eb bees as well as between Eb and Ea bees. Whereas that between Ee and Eb bees was longer. Further analysis indicated that the allele frequency of seven alleles at six loci (159 bp at A29,100 bp and 104 bp at A24; 110 bp at A7; 126 bp at A43, 221 bp at A14 and 221 bp at A113) increased going from Ee to Eb to Ea bees. Paired tests showed significant higher allele frequency between Ea and Eb bees,as well as between Ea and Eb bees. This indicates that these seven alleles are likely molecular markers of the high royal jelly producing bees. In addition,the allele frequency of four alleles at four loci (106 bp at A24,140 bp at A43;215 bp at A113 and 219 bp at A14) decreased going from Ea bees to Eb to Ee. Paired tests indicated significant lower allele frequency between Ea and Ee bees,as well as between Ea and Eb bees. Those four alleles may be the genetic markers for low royal jelly production. PMID- 16252699 TI - [Expressed sequence tags (ESTs) analysis of the oral gland of Lampetra japonica]. AB - A cDNA library (2.1 x 10(6) pfu/mL) was constructed from the oral gland of Lampetra japonica. After random selection of clones for sequencing, 1 323 clones with inserts longer than 100 bp and with good chromato-grams were obtained. Using BlastX and BlastN programs, We found 653 ESTs (49.36%) that shared significant homology with known sequences in protein or nucleotide databases of NCBI, including 328 ESTs homologous to known sequences of Petromazonidae animals. These ESTs were classified into 11 different functional categories,the highest proportion of which was related to protein synthesis. Out of the 1323 ESTs, 162 non-redundant contigs were assembled from 547 sequences after alignment using software at the threshold of more than 90% homology over a minimum of 20 base pairs. There were eight full-length cDNAs with complete open reading frame. Further studies on the ESTs in the library may be helpful to elucidate the components of Lampetra japonica oral gland and to understand the function of those proteins at the molecular level. PMID- 16252700 TI - [Genetic differentiation of domestic goose breeds in China]. AB - The 1 042 bp control region of mitochondrial DNA from 84 geese of 15 domestic goose breeds was sequenced and genetic differentiation was analysed. Results showed that the interpopulation nucleotide divergence was highest (3.805% 4.067%) between Yili and the other 14 breeds. The average nucleotide diversity variation within different domestic breeds was 0 - 0.116%. Excluding the Yili, the interpopulation nucleotide divergence between Huoyan and the remaining breeds, was 0.211% - 0.272%, which was significantly higher than that between any other two breeds (0 -0.094%). During the formation of domestic breeds in China,there is an association between the genetic differentiation of domestic geese and their geographic distribution. The divergence time of Huoyan breed was relatively earlier and genetic drift may have been the main factor to affect the genetic differentiation of the Huoyan breed (Nm = 0.02 -0.54). On the other hand, gene flow is the main reason for the lack of a clear differentiation among the remaining 13 Chinese domestic geese breeds (Nm = 12.0 - 65.33). PMID- 16252701 TI - [Estrogen receptor as a candidate gene for prolificacy of small tail Han sheep]. AB - Single nucleotide polymorphism in exon 1 of the estrogen receptor (ESR) gene was detected by PCR-SSCP in both high fecundity sheep breeds (Small Tail Han sheep, Hu sheep and German Mutton Merino sheep) and low fecundity sheep breeds (Dorset sheep,Suffolk sheep). Results indicated that there were three genotypes (AA, AB and BB) in all three high fecundity sheep breeds, but only two genotypes (AA, AB) in both low fecundity breeds. In Hu sheep,German Mutton Merino sheep, Small Tail Han sheep, Suffolk sheep and Dorset sheep,the frequency of allele A was 0.672, 0.786, 0.846, 0.857 and 0.867, respectively, and the frequency of allele B was 0.328, 0.214, 0.154, 0.143, and 0.133, respectively. Sequencing revealed a C-->G mutation at 363 bp of exon 1 of ESR gene in the BB genotype in comparison to the AA genotype. The genotype distribution was significantly different between Small Tail Han sheep and Hu sheep (P<0.01) and between Dorset sheep and Hu sheep (P <0.05). There was no difference in genotype distribution between other sheep breeds. The Small Tail Han sheep ewes with genotypes AB or BB had 0.51 (P < 0.05) and 0.7 (P < 0.05) more lambs than those with genotype AA, respectively. These results showed that the estrogen receptor locus is either a major gene that influences the prolificacy in Small Tail Han sheep or in close linkage with such a gene. In view of our results, marker-assisted selection using ESR is warranted to increase litter size in sheep and will be of considerable economic value to mutton producers. PMID- 16252702 TI - QTL for yield and its components responded to elevated CO2 in rice (Oryza sativa L.). AB - FACE (free air carbon dioxide enrichment) technology may provide a means by which the environment around growing plants can be modified to realistically simulate the concentration of atmospheric CO2 in the future. The plant growth and its yield of plant species can be enhanced under FACE. Identification of genomic regions influencing the response of yield and its components to elevated CO2 will be useful for understanding the genetics of active response to changed CO2 environment and developing higher yield cultivars, which will be adapted to future enriched atmospheric CO2 environment. A mapping population of 65 indica (IR24) chromosome segment substitution lines (CSSLs) in japonica (Asominori) background and their parents were used to detect QTLs for yield and its components, e. g. number of fertile tillers per plant (FT), 1000-grain weight (TGW), number of grains per panicle (GP) and grain yield per plant (GY) under FACE (200 micromol CO2/mol above current levels) and current CO2 concentration (Ambient, about 370 micromol CO2/mol) in the field experiment. The results showed that, GY, GP and FT of two parents under FACE were significant greater than that under Ambient. The transgressive segregation of the four traits was observed in the CSSLs population under both FACE and Ambient. A total of 20 QTLs for the four traits were detected on chromosome 1, 2, 4, 6, 7, 9 and 12 with LOD (Log10 likelihood ratio) of QTLs ranging from 2.5 to 5.7. Three QTLs were detected under both FACE and Ambient. However,other QTLs were detected only under one level of CO2, which indicated that these loci were sensitive to CO2 concentration. Additionally, two QTLs qFT12 and qGP4 were found for the QTL x Environment (QE) interaction effects. It is suggested that there is a high possibility to improve the yield of rice under elevated CO2 through marker-assisted selection. PMID- 16252703 TI - [Study on early stability by ISSR markers in rice]. AB - In the F2 population crossed from two early stability rice (Oryza sativa L.) with four cultivars, eight uniform strains were recorded. Genetic analysis showed that both uniform strains with uniform agronomic characteristics and segregated strains segregating in Mendelian manner were observed in F2 population of the same combination. Four types of marker-bands were obtained after the F2 uniform strains were marked by 26 ISSR primers:t ype I, maternal marker bands present, and paternal marker bands absent; type II, paternal marker bands present, and maternal marker bands absent; type III, parts of maternal and paternal marker bands present, and the others absent; type IV, new recombined marker bands appeared, and all maternal and paternal marker bands absent. But segregating strains showed heterzygosity marker bands, maternal and paternal marker bands being present. The uniform strains and normal strains in rice might be grouped into two classes on the basis of the 2000 bp marker band amplified by the ISSR marker primers No. 900. This result would provide experimental basis for the study on genetic mechanism of early stability in rice. PMID- 16252704 TI - [Generation of chemical-inducible activation tagging T-DNA insertion lines of Arabidopsis thaliana]. AB - Using an estrogen-inducible expression XVE (LexA-VP16-Estragon Receptor) system, we have generated approximately 40 000 independent T-DNA insertion lines of Arabidopsis thaliana. Segregation analyses of about 18000 lines indicated that 51.6% of them contain single T-DNA insertions and that the average insertion number is 1.38 copies per line. Mutants displaying a variety of morphological alterations were identified, including those that affect development of roots,hypocotyls, leaves, floral organs and seeds as well as the flowering time. PMID- 16252705 TI - [Genetic contribution of agronomic traits to yield in flue-cured tobacco (Nicotiana tabacum L.)]. AB - In order to understand the genetic contribution of six agronomic traits to yield, 14 flue-cured tobacco varieties (or breeding lines) and their 41 F1 crosses were used for multivariable conditional analysis. The contribution of additive variance of plant height to yield was larger than other agronomic traits. The largest contribution of dominant variance to yield was due to the length of middle leaves. All agronomic traits investigated had small contribution to yield due to additive x environment interaction effects and dominant x environment interaction effects. No identical trait of different parents showed the largest contribution to additive effect of yield. This could be resulted from the fact that each parent had its own genetic and developmental characterization. The dominant effects of yield were mainly influenced by length of middle leaves in most crosses. Length of middle leaves could be served as ameasurement to indirectly select the cross parent having high dominant effect of yield. PMID- 16252706 TI - [Comparative analysis of sequences of the 5S rDNA NTS in wild close relatives of barley from Tibet of China]. AB - Tibet is a center of distribution and differentiation of genus Hordeum in China,and has a great deal of species resource. The sequences of the nontranscribed intergenic spacer (NTS) region of 5S nuclear ribosomal DNA was studied in 18 varieties of the close relative of wild barley (Hordeum spontaneum) from different geographical regions in Tibet and Turkmenistan. These sequences were determined by sequencing the clones of PCR products. Alignment of sequences revealed that the 5S rDNA NTS contained two comparatively conservative regions, A and B, and a variable TAG repeat (V). The number of TAG repeats varies from 4-17, also including several transitions and conversions (TAG-->TCG, TAG-->TAC). The total size of the conservative regions was from 168 -169 bp, and the sequence length variation was only 1 bp. The GC content (%) of the conservative sequence was 43.8% and the homologous of that nearly 98.2% -100%. The number of variable sites was 12 (7.1%). In general, there were more transitions than conversions in the variation sites,and the ratios of transition/conversion were 1.0 - 2.0. NTS polymorphism of 5S rDNA was mainly determined by polymorphism of TAG repeats. The molecular system-tree showed that the NTS should be a useful marker to classify the accessions in Hordeum. PMID- 16252707 TI - CPD banding patterns and identification of 45S rDNA sites in tomato. AB - In this study, we performed sequentially combined PI and DAPI (CPD) staining and FISH with two different 45S rDNA clones on meiotic pachytene and mitotic metaphase chromosomes in tomato. Ten red CPD bands were shown on eight pachytene bivalents, and 12 bands were shown on six pairs of mitotic metaphase chromosomes. The CPD bands exhibited on mitotic metaphase chromosomes corresponded to the prominent bands exhibited on the pachytene chromosomes. The distinctive CPD bands, which could be constantly and clearly detected using the CPD staining procedure we improved, provided new landmarks for chromosome identification in tomato. FISH with the tomato 45S rDNA clone revealed very strong signal(s) in the satellite(s) on the short arm of chromosome 2 as well as weak signals in five CPD banded regions at pachytene or four pairs of CPD banded regions at metaphase chromosomes. However, FISH with pTa71 plasmid only revealed signals in the satellite. Considering the difference in sequence between the two rDNA clones, we inferred that only the satellite contains the coding regions of 45S rDNA unit in tomato. The property of CPD bands as well as the DNA sequences probably involved in the five CPD banded regions was discussed. PMID- 16252708 TI - [Progress on genetic susceptibility to ankylosing spondylitis]. AB - Ankylosing spondylitis (AS) is a highly heritable, common inflammatory rheumatic disease. Since 1970s, evidence has increasingly pointed to HLA-B27 as the major gene involved in susceptibility to AS. However, genome-wide scan and association studies have shown, in addition to HLA, regions outside the HLA are involved in susceptibility to AS. Furthermore, there is compelling evidence that non-B27 genes, either within or outside the HLA, are involved in disease aetiology. This review mainly summarized different genes associated with AS, and the current progress in this exciting area. PMID- 16252709 TI - The NHS plan. PMID- 16252710 TI - A clean sweep. PMID- 16252711 TI - Send in the clowns. PMID- 16252712 TI - Winning the lottery. PMID- 16252714 TI - Partners in care. PMID- 16252713 TI - Hope for the future. PMID- 16252715 TI - Spirituality and religion: exploring the relationship. PMID- 16252716 TI - The evidence base for spiritual care. PMID- 16252717 TI - Managing multiple sclerosis: working in partnership. PMID- 16252718 TI - [Effect of rare-earth element Eu3+ on callus growth and flavonoids content in Glycyrrhiza uralensis]. AB - The effect of rare-earth element Eu3+ on callus growth and its flavonoids content in Glycyrrhiza uralensis was studied. The results showed that lower concentration of rare-earth element Eu3+ could promote the callus growth and flavonoids productions. 0.1 mg/l Eu3+ was the most suitable for the biomass accumulation of the callus and flavonoids biosynthesis. In that concentration flavonoids content was 2.7 times and liquiritigenin was 4 times as that of the control. PMID- 16252719 TI - [Pharmacognostic identification of dried twigs and leaves of Taraphochlamys affinis (Giff) Bremekhu]. AB - The pharmacognosy of Taraphochlamys affinis (Giff) Bremekhu was studied by microscopic observation to provide a scientific basis for the identification, development and utilization of its resources. As a result, obvious characteristics for its identification were revealed, which could be used to identify twigs and leaves of Taraphochlamys affinis (Giff) Bremekhu. PMID- 16252720 TI - [Biphenyl ferulate from Glehnia littoralis]. AB - OBJECTIVE: To investigate the prolyl oligopeptidase inhibitory constituents from Glehnia littoralis. METHODS: To determine the prolyl oligopeptidase inhibitory activity in vitro of ethanol extract of G. littoralis and its fractions with Yoshimoto's method, to separate the constituents of the active fractions by repeated silica gel column chromatogrphy. RESULTS: Two ferulate compounds were isolated from EtOAc-soluble fractions of ethanol extract of G. littoralis, which (100 microg/ml) showed mile activity (inhibitory rate: 50%). Compound I, named glehnilate, was a new biphenyl ferulate. Compound II, identified as 2-(4' hydroxyphenol)-glycol mono trans-ferulate, was isolated from this plant for the first time. Their structures were determined on the basis of their spectral data. PMID- 16252721 TI - [Chemical constituents from Corydalis humosa]. AB - OBJECTIVE: To study the chemical constituents of Corydalis humosa Migo. METHODS: Some chromatography methods were used to separate the chemical constituents of this plant. Their structures were determined by NMR and MS spectral data. RESULTS: Six compounds were isolated from the 95% ethanolic extract of the tubers of C. humosa. The structures of these compounds were identified as tetrahydropalmatine (1), jatrorrhizine (2), berberine (3), tetrahydroberberine (4), palmaline (5) and (-)-tetrahydrocoptisine (6) respectively. CONCLUSIONS: Compounds 1-3 were isolated from this plant for the first time. PMID- 16252722 TI - [Studies on the triterpenoids of Cyclocarya paliurus (Batal.) Iljinsk]. AB - Three triterpenes (I-II) were obtained from the leaves of Cyclocarya paliurus (Batal.) Iljinsk. By means of physicochemical and spectral methods, the structures of the three triterpenes were identified as oleanolic acid (I), ursolic acid (II) and epikatonic acid (III) respectively. All of the three triterpenes were isolated for the first time from the plant of Cyclocarya paliurus (Batal.) Iljinsk. PMID- 16252723 TI - [Effect of penetration enhancers on the transdermal penetration of sinomenine liposome patch]. AB - OBJECTIVE: To study the effect of penetration enhancers on the transdermal penetration of sinomenine liposome patch. METHODS: We firstly established the experimental method of transdermal penetration, and the methodology study was carried out at the same time. Furthermore, we added 5% Azone, 10% Oleic acid, 1% PG + 4% Azone, 2% PG + 8% Oleic acid into the patch. The cumulative penetration amounts were determined during the test of transdermal penetration and the part lagged in the skin was determined after the test. RESULTS: The cumulative penetration amounts released from the patch increased, while the part lagged in the skin decreased at the same time. CONCLUSIONS: The results indicate that penetration enhancers can facilitate drug through corneum into dermis, but can't enhance the drug' s accumulation in the corneum. It is concluded that penetration enhancers can destroy the two-molecule structure of liposomes and cause drug leakage, so penetration enhancers should not be added into liposome patch of part functions. PMID- 16252724 TI - [Study on proliferation effect of extracts of Piper longum on mesenchymal stem cells of rat bone marrow and the relationship to chemical functional groups]. AB - OBJECTIVE: To observe the effect of volatile oil and aqueous soluable part of Piper longum on proliferation of rat mesenchymal stem cell (MSC) and the chemical functional groups. METHODS: Mesenchymal stem cells were dissociated from rat bone marrow and marked by Brdu, and the expression of CD44, CD54 and double label of Brdu and CD44. The growth of rat mesenchymal stem cell under volatile oil and aqueous soluable part of Piper longum was observed by means of cell viability measurement (MTT) and morphological observation and Brdu, PCNA immunohistochemical methods. RESULTS: Volatile oil of Piper longum could promote the cell viability of MSC and the number of Brdu, PCNA positive cell in dose dependant. There was significant difference in comparision with control groups, C = C(50.66%), -OH(27.02%) and other functional groups in volatile oil of Piper longum were determined by GC-MS, but the aqueous soluable part of Piper longum could not promote the proliferation of MSC. CONCLUSIONS: Volatile oil of Piper longum which consists of C = C, -OH, and other functional groups has strong effect on enhancing proliferation of MSC. PMID- 16252725 TI - [Inhibitory effect of ardipusilloside-I on Lewis pulmonary carcinoma and hepatocarcinoma SMMC-7721]. AB - OBJECTIVE: To study the antitumor effect of ardipusilloside-I (ADS-I) on Lewis pulmonary carcinoma and hepatocarcinoma SMMC-7721. METHODS: Lewis pulmonary metastasizing model was made by Sc. diluted 3-time 20 microl cancer cells in the right sole of C57BL/6 mice, after we treated the mice 14 days with ADS-I orally, cut the neoplasia-foot and weighed, we calculated the inhibitory rate in situ, then put the mice to death after being administrated 11 days continuously, counted the lung medtastasis colony and calculated the metastasis inhibitory rate; Human being hepatocarcinoma model was made by sc. in the back 5 x 10(6)/ml SMMC-7721 cells per BALB/c/nu nude mice, after we treated them orally 16 days with ADS-I continuously, measured the volume of tumor growth and body weight, and drew the growth curve, we detached the tumor and calculated the inhibitory rate of tumor growth in the end. RESULTS: Lewis pulmonary carcinoma inhibitory rate in situ and lung transfer inhibitory rate of ADS-I (25 mg/kg - 100 mg/kg) were between 40.5% and 54.0%, 45.4% and 69.1% respectively; Inhibitory rate of hepatocarcinoma SMMC-7721 was between 45.6% and 56.3%. CONCLUSIONS: ADS-I has inhibitory effect on Lewis pulmonary metastasis carcinoma and hepatocarcinoma SMMC-7721 carcinoma. PMID- 16252726 TI - [Effect of Ganoderma triterpene on proliferation of dendritic cells from mouse spleen]. AB - OBJECTIVE: To investigate the effect of Ganoderma Triterpene (GT) on proliferation of Dendritic cells (DC) from mouse spleen. METHODS: Compared with the cytokine (GM-CSF + IL-4), we investigated the effect of GT of different concentrations and cytokine + GT of different concentrations on proliferation of DC from mouse spleen by MTT. RESULTS: GT (40-200 microg/ml) could stimulate the proliferation of DC significantly, but the stimulations weren't obvious compared with the cytokine. GT + cytokine, compared with the negative control, had obviously promoting effect and was better than the cytokine. The results above indicated GT could not only stimulate the proliferation of DC but also significantly coorderate with the cytokine. CONCLUSIONS: GT may play the role by its growth factor-like function, and by cooperating with the cytokine in the regulation of DC. PMID- 16252727 TI - [Enhancement of wound healing by taspine and its effect on fibroblast]. AB - OBJECTIVE: To study the effect of taspine on enhancement of skin wound healing and its effect on fibroblast proliferation and autocrine. METHODS: The plerosis effect of taspine on experimental skin wound was observed in vivo. Different concentrations of taspine were added in vitro and MTT technique was applied to observe its effect on fibroblast proliferation, the levels of transforming growth factor-beta1 (TGF-13P) and epidermal growth factor (EGF) were determined by ELISA. RESULTS: In vivo, exo-applied taspine 300 microg and 150 microg accelerated the recovery of skin wound. In vitro, 0.50-0.4 microg/ml taspine could increase autocrine of TGF-beta1and EGF by fibroblast, but it showed no effect on L929 fibroblast proliferation. CONCLUSIONS: Taspine enhances wound healing by increasing the autocrine of TGF-beta1 and EGF by fibroblast. PMID- 16252728 TI - [Experimental study of Oleanolic acid liquid acting on climacteric rats]. AB - OBJECTIVE: To study whether the Oleanolic acid had curative effect on climacteric rats as an independent factor. METHODS: The experimental group was administrated with Oleanolic acid, while the positive group was given with Isoflavniphytoestrogeno, and the climacteric model and adolescent control groups were fed with 0.9% saline water. The effect of Oleanolic acid on rats was observed. In each group the levels of serum E2, SOD, MDA, GSH-Px were tested, the morphology of ovary and adrenal cortex were also studied. RESULTS: It was showed that Oleanolic acid could increase the levels of serum E2, SOD, GSH-Px (P < 0.05), decrease the level of MDA (P < 0.01) and improve the morphology and the functions of ovary and adrenal cortex. CONCLUSIONS: It is concluded that Oleanolic acid has curative effect independently on climacteric rats. PMID- 16252729 TI - [Determination of amino acids in fu fang banlangen granulae by a pre-column derivatization RP-HPLC method]. AB - OBJECTIVE: To establish a pre-column derivatization reversed-phase high performance liquid chromatographic method for the determination of amino acids, which was known as the characteristic constituents of Fu Fang Banlangen Granulae. METHODS: We determined the amino acids in alkalescence by a pre-column derivatization of DNFB. NaAc buffer (pH 6.4)-Acetonitrile (850:150) was used as the mobile phase on a SinoChrom ODS-BP column. The detector was operated at 360 nm. RESULTS: The method was carried out over the range of 0.627 - 5. 016 microg for Arg and 0.874 - 7.000 microg for Pro. The recoveries were 97.7% with the RSD = 2.0% for Arg and 97.7% with the RSD = 1.5% for Pro. CONCLUSIONS: The method has good accuracy and repeatability and it can be used for the quality control of Fu Fang Banlangen Granulae. PMID- 16252730 TI - [Study of HPLC chromatographic fingerprint of Citrus reticulata Blanco cv. Dahongan]. AB - OBJECTIVE: To establish HPLC chromatographic fingerprint of Citrus reticulata Blanco cv. Dahongan. METHODS: The experimental condition of the RP-HPLC method was as follows: Angilent Zorbax C18 column (4.6 mm x 250 mm, 5 microm), with linear gradient elution using (A) methanol and (B) water; The flow rate was 1.0 ml/min, and the injected volume was 20 microl, detected wavelength was 283 nm. RESULTS: Under the qualitative conditions we determined different samples of Citrus reticulata Blanco cv. Dahongan. There were 11 common peaks in chromatographic. CONCLUSIONS: According to analyze and contrast the relative retention values and relative peak area in chromatographic fingerprint, we establish HPLC chromatographic fingerprint of Citrus reticulata Blanco cv. Dahongan to provide a scientific basement for the quality evaluation of it. PMID- 16252731 TI - [Applying grading methods of synthesizing multiple guidelines to optimizing extracting technology for Radix Panacis Quinquefolii]. AB - OBJECTIVE: To optimize the best extracting procedure for Radix Panacis Quinquefolii. METHODS: (1) The optimum alcohol extracting procedure was selected with the content and extraction efficiency of ginsenoside Rg, Re, Rb, from Radix Panacis Quinquefolii by orthogonal test design and synthesizing multiple guidelines method. (2) The content of three ginsenoides Rg1, Re and Rb1 in Radix Panacis Quinquefolii was determined by gradient elution method in RP-HPLC. RESULTS: The optimum alcohol-extracting procedure was determined as 12 times of 70% alcohol refluxing and extracting 3 h for 2 times. CONCLUSIONS: The optimum extracting procedure can increase extraction efficiency of Radix Panacis Quinquefolii. PMID- 16252732 TI - [Determination of oxymatrine content and its related substances in oxymatrine capsule by HPLC]. AB - OBJECTIVE: To determine oxymatrine content and its related substances in oxymatrine capsule. METHODS: A HPLC method was adopted. C18 column was used. The mobile phase consisted of phosphate buffer solution-methanol (85:15). The UV detective wavelength was 210 nm. RESULTS: The calibration curve of oxymatrine was linear in the range of 6.952 - 20.856 microg/ml (r = 0.9999). The calibration curve of N-oxysophocarpine was linear in the range of 0.1837 - 0.5510 microg/ml (r = 0.9997). CONCLUSIONS: The method is simple, rapid and accurate. PMID- 16252733 TI - [The preparation and quality control of Solanum lyratum Thunb Extract gel]. AB - OBJECTIVE: To introduce the preparation and quality control of Solanum lyratum Extract Thunb gel. METHODS: The gel was prepared with Solanum lyratum Thunb Extract as main component and carbopol 940 as base material, and Solasodine was determined by TLCS. RESULTS: The preparation of this gel was feasible, its quality was reliable, its detection method was rapid, stable, and could be used to control the quality of Solanum lyratum Thunb Extract gel. CONCLUSIONS: Studies on Solanum lyratum Thunb Extract gel will find a new path for the treatment of eczema with mono traditional Chinese medicine. PMID- 16252734 TI - [Clinical observation of the effect of xinmaitong tablet on hypertension]. AB - OBJECTIVE: To observe the clinical therapeutic effect of Xinmaitong on hypertension. METHODS: 200 primary hypertension patients were assigned to Xinmaitong group and Nimodipine group randomly. Compare the effective rate between two groups after 6 weeks' therapy. RESULTS: The degree of decreasing blood pressure of Xinmaitong group was obviously higher than Nimodipine group, there was significant difference between them (P < 0.05). CONCLUSIONS: The effect to therapy hypertension of Xinmaitong is better than Nimodipine, and there is no obvious adverse effect. PMID- 16252735 TI - [Clinical study on effect of matrine injection to protect the liver function for patients with primary hepatic carcinoma after trans-artery chemo-embolization (TAE)]. AB - OBJECTIVE: To study effect of Matrine injection to prevent the liver function of patients with primary hepatic carcinoma (PHC) after trans-artery chemo embolization. METHODS: 122 patients with PHC admitted to our hospital between October 1999 and June 2004 were randomly divided into therapy group (62 cases) and control group (60 cases). In therapy group, the patients after trans-artery chemo-embolization (TAE) were infused 150 mg Matrine injection once a day for two weeks, while we used other same hepatinica on patients in control group. The levels of alamine aminotransferase (ALT) , aspartate aminotransferase (AST), serum total bilirubin (STB), Albumin (ALB) were tested before and after one week and two weeks of TAE. RESULTS: In therapy group, the levels of ALT and AST at one week after TAE were much higher than that before TAE (P <0.01) until two weeks later (P > 0.05). In control group, the levels of ALT, AST, STB were all elevated at one week (P < 0.01), and that of the ALT and AST did not come down to the levels at two weeks before TAE. CONCLUSIONS: Matrine injection may be used to protect the liver function for patients with PHC after TAE, to relieve the liver cells damage, and to improve the tolerance of TAE, so as to perform the next TAE in time. PMID- 16252736 TI - The increasing use of alcohol-based hand sanitizers. PMID- 16252737 TI - Office-based anesthesia: requirements for patient safety. PMID- 16252738 TI - Factors influencing the discomfort of intraoral needle penetration. AB - This study with 24 volunteers compared the discomfort produced by needle penetration in different parts of the palatal mucosa. In addition, comparing a fresh needle to one that was used for a previous penetration in the same patient, we assessed the influence of the status of the needle on insertion discomfort during buccal mucosal penetration. The results showed that needle penetration in the anterior hard palate was more uncomfortable than in the posterior palate. Although men could not differentiate between fresh and used needles for a second buccal mucosal penetration, women reported a significant increase in discomfort with used needles. PMID- 16252739 TI - Initial injection pressure for dental local anesthesia: effects on pain and anxiety. AB - This study quantitatively assessed injection pressure, pain, and anxiety at the start of injection of a local anesthetic into the oral mucosa, and confirmed the relationship between injection pressure and pain, as well as between injection pressure and anxiety. Twenty-eight healthy men were selected as subjects and a 0.5-inch (12 mm) 30-gauge disposable needle attached to a computer-controlled local anesthetic delivery system (the Wand) was used. A 0.5 mL volume of local anesthetic solution was injected submucosally at a speed of either 30 or 160 s/mL. Three seconds after the start of local anesthetic injection, injection pressure was measured and pain and anxiety were assessed. Injection pressure was measured continuously in real time by using an invasive sphygmomanometer and analytical software, and pain was assessed on the Visual Analogue Scale and anxiety on the Faces Anxiety Scale. A significant correlation was evident between injection pressure and pain (rs = .579, P = .00124) and between intensity of injection pressure and state anxiety (rs = .479, P = .00979). It is therefore recommended that local anesthetic be injected under low pressure (less than 306 mm Hg) to minimize pain and anxiety among dental patients. PMID- 16252741 TI - Enhancement of stimulated Brillouin scattering of higher-order acoustic modes in single-mode optical fiber. AB - Solving the elastic wave equation exactly for a GeO2-doped silica fiber with a steplike distribution of the longitudinal and shear velocities and density, we have obtained the dispersion, attenuation, and fields of the leaky acoustic modes supported by the fiber. We have developed a model for stimulated Brillouin scattering of these modes in a pump-probe configuration and provided their Brillouin gains and frequencies for an extended range of core sizes and GeO2 doping. Parameter ranges close to cutoff of the acoustic modes and pump depletion enhance the ratio of higher-order peaks to the main peak in the Brillouin spectrum and are suitable for simultaneous strain-temperature sensing. PMID- 16252740 TI - Tiagabine may reduce bruxism and associated temporomandibular joint pain. AB - Tiagabine is an anticonvulsant gamma-aminobutyric acid reuptake inhibitor commonly used as an add-on treatment of refractory partial seizures in persons over 12 years old. Four of the 5 cases reported here indicate that tiagabine might also be remarkably effective in suppressing nocturnal bruxism, trismus, and consequent morning pain in the teeth, masticatory musculature, jaw, and temporomandibular joint areas. Tiagabine has a benign adverse-effect profile, is easily tolerated, and retains effectiveness over time. Bed partners of these patients report that grinding noises have stopped; therefore, the tiagabine effect is probably not simply antinociceptive. The doses used to suppress nocturnal bruxism at bedtime (4-8 mg) are lower than those used to treat seizures. PMID- 16252742 TI - Thermo-optically tunable band-rejection filters using mechanically formed long period fiber gratings. AB - We present tunable mechanically formed long-period fiber gratings whose resonance peaks are shifted by the thermo-optic effect of a medium surrounding the fiber cladding. A novel grating formation method and tuning principles that use periodically arrayed metal wire are described. The performance as an active gain equalizer of an erbium-doped fiber amplifier is also reported. PMID- 16252743 TI - Wideband all-order polarization mode dispersion compensation via pulse shaping. AB - We demonstrate the application of ultrafast pulse-shaping techniques for experimental wideband all-order polarization mode dispersion (PMD) compensation, for the first time to our knowledge. PMD is treated as arbitrary variations of state of polarization and phase versus wavelength, in an all-order sense. Consequently, two pulse shapers are implemented in a serial manner to compensate for the polarization and the phase spectra independently. We report compensation of subpicosecond pulses (14 nm bandwidth around 1550 nm) that are anomalously spread to more than 2 ps as a result of PMD. This PMD compensation scheme can potentially be a powerful and cost-effective solution for fiber optic telecommunication networks. PMID- 16252744 TI - 1.1 MW peak-power, 7 W average-power, high-spectral-brightness, diffraction limited pulses from a photonic crystal fiber amplifier. AB - We report a large-core, Yb-doped photonic crystal fiber amplifier generating diffraction-limited (M2 = 1.04), approximately 0.45 ns, approximately 8 GHz linewidth pulses with energies of 540 microJ, peak power in excess of 1.1 MW, and average power of approximately 7.2 W at a repetition rate of 13.4 kHz. The pulse peak spectral brightness exceeds 10 kW/(sr Hz cm2) and is the highest generated by a fiber source, to our knowledge. PMID- 16252745 TI - Multiport N x N multimode air-clad holey fiber coupler for high-power combiner and splitter. AB - We report 2 x 2 and 4 x 4 multimode air-clad holey fiber couplers fabricated by a novel fusion and tapering technique. The devices showed excellent port-to-port coupling uniformity over a wide spectral range of 800-1650 nm. The 4 x 4 coupler showed a low insertion loss of 9.9 dB and excess loss of 3.9 dB at 1310 nm. Because of its unique air cladding and the high numerical aperture of the holey fiber structure, the device showed strong potential in high-power applications. PMID- 16252746 TI - Long-period fiber gratings as ultrafast optical differentiators. AB - It is demonstrated that a single, uniform long-period fiber grating (LPFG) working in the linear regime inherently behaves as an ultrafast optical temporal differentiator. Specifically, we show that the output temporal waveform in the core mode of a LPFG providing full energy coupling into the cladding mode is proportional to the first derivative of the optical temporal signal (e.g., optical pulse) launched at the input of the LPFG. Moreover, a LPFG providing full energy recoupling back from the cladding mode into the core mode inherently implements second-order temporal differentiation. Our numerical results have confirmed the feasibility of this simple, all-fiber approach to processing optical signals with temporal features in the picosecond and subpicosecond ranges. PMID- 16252747 TI - Tunable highly birefringent photonic bandgap fibers. AB - A novel tunable highly birefringent photonic bandgap fiber (PBGF) is designed theoretically by filling its air holes with high-index material. The transmission band can be continuously tuned by changing the refractive index of the filling material. Accordingly, the tunable modal birefringence and polarization mode dispersion of the PBGFs are investigated by adjusting the refractive index of the filling material. Furthermore, we have also analyzed the effect of surface modes in the photonic bandgap on the characteristics of the tunable PBGFs. The simulation results show the feasibility of constructing birefringence-tunable photonic crystal fibers and related fiber devices in practical applications. PMID- 16252748 TI - Recovering correct phase information in multiwavelength digital holographic microscopy by compensation for chromatic aberrations. AB - We demonstrate experimentally that correct phase imaging without 2pi ambiguity is obtainable in digital holography by using a multiwavelength approach in the microscope configuration. We describe a general approach for removing chromatic aberrations and for controlling the pixel size of the reconstructed phase image in multiwavelength digital holography when the Fourier transform method is adopted for the numerical reconstruction of digital holograms. The retrieved phase is affected by the unavoidable, unwanted chromatic aberration. The correct phase can be obtained by evaluating the phase from the reference holograms reconstructed at different wavelengths to compensate for the chromatic aberration. PMID- 16252749 TI - Sensitivity variation in two-center holographic recording. AB - An experimental study of variation of sensitivity with recording and sensitizing intensities in two-center recording is presented. The experimental results are in good agreement with the theoretical predictions. It is shown experimentally, for what is to our knowledge the first time, that the sensitivity is a function of the ratio of recording to sensitizing intensities and not the absolute intensities. Also, the ratio of recording to sensitizing intensities should be small to obtain high sensitivity values. We also report the highest sensitivity (S=0.15 cm/J) that has been achieved to date for a LiNbO3:Fe:Mn crystal. PMID- 16252750 TI - Laser-tweezer-controlled solid immersion microscopy in microfluidic systems. AB - We describe the creation and implementation of a near-field scanning solid immersion microscope that is specifically tailored for use in microfluidic systems. The microscope comprises a newly fabricated Weierstrass solid immersion lens (SIL), which is detached from its substrate and is free floating in the fluid, and a laser optical tweezer, which serves both as a trapping beam for alignment and positioning of the SIL and as a near-field scanning beam that images the sample through the SIL. A discussion of the SIL's fabrication method is presented along with experimental results that demonstrate the effectiveness of our microscope design. PMID- 16252751 TI - Decomposition of the optical transfer function: wavefront coding imaging systems. AB - We describe the mapping of the optical transfer function (OTF) of an incoherent imaging system into a geometrical representation. We show that for defocused traditional and wavefront-coded systems the OTF can be represented as a generalized Cornu spiral. This representation provides a physical insight into the way in which wavefront coding can increase the depth of field of an imaging system and permits analytical quantification of salient OTF parameters, such as the depth of focus, the location of nulls, and amplitude and phase modulation of the wavefront-coding OTF. PMID- 16252752 TI - Optical edge-projection method for three-dimensional profilometry. AB - A novel optical edge-projection method is proposed for surface contouring of an object with low reflectivity. A structured light edge is projected onto a dark surface, and the image is captured by a CCD camera. The object contour is evaluated with an active triangular projection algorithm, and one obtains a whole field three-dimensional contour of the object by scanning the optical edge over the entire object surface. The proposed method is applied to a black nonreflective object made from woven carbon fiber and is also applied to measure the profile of a small object (a coin). The results show that an accurate profile of the specimen can be obtained. PMID- 16252753 TI - Compact multireference wavefront sensor design. AB - We present a compact optical design for a multireference Shack-Hartmann-based wavefront sensor (WFS) for multiconjugate adaptive optical systems. The key component of this WFS design is a field lenslet array that separates the exit pupil images in the sensing plane for all reference sources. An analytical method for WFS optical design is presented, and the optimal strategy for selecting optical components from a discrete set is outlined. The feasibility of the WFS design has been demonstrated for a prototype WFS system in a laboratory setup with five reference sources and two deformable mirrors representing a wavefront distorting medium. PMID- 16252754 TI - Optimal design of optical reference signals by use of a genetic algorithm. AB - A new technique for the generation of optical reference signals with optimal properties is presented. In grating measurement systems a reference signal is needed to achieve an absolute measurement of the position. The optical signal is the autocorrelation of two codes with binary transmittance. For a long time, the design of this type of code has required great computational effort, which limits the size of the code to approximately 30 elements. Recently, the application of the dividing rectangles (DIRECT) algorithm has allowed the automatic design of codes up to 100 elements. Because of the binary nature of the problem and the parallel processing of the genetic algorithms, these algorithms are efficient tools for obtaining codes with particular autocorrelation properties. We design optimum zero reference codes with arbitrary length by means of a genetic algorithm enhanced with a restricted search operator. PMID- 16252755 TI - High-resolution angular measurement using surface-plasmon-resonance via phase interrogation at optimal incident wavelengths. AB - It is demonstrated that ultrahigh-resolution angular measurement can be achieved via surface-plasmon-resonance excitation in which the phase difference between p- and s-polarized reflected waves is monitored as a function of the incidence angle. Resolutions down to 1.9 x 10(-6) deg are obtained by performing the measurements at optimal incident wavelengths. This represents an order of magnitude improvement compared with previously reported values. PMID- 16252756 TI - 157 nm F2-laser writing of silica optical waveguides in silicone rubber. AB - Silica (SiO2) optical waveguides have been fabricated on the surface of silicone [(SiO(CH3)2)n] rubber by photochemical modification of silicone rubber into silica with 157 nm F2-laser radiation. The 2 mm thick silicone was exposed through a thin (approximately 0.2 mm) air layer to generate oxygen radicals that chemically assisted in the silica transformation. Silica waveguides were defined in 8-16 microm wide exposure strips by a proximity Cr-on-CaF2 photomask. Optimum laser processing conditions are presented for generating crack-free waveguides with good optical transparency at red (635 nm) and infrared (1550 nm) wavelengths. A propagation loss of approximately 6 dB/cm is reported at the 1550 nm wavelength. PMID- 16252758 TI - Three-color photonic crystal demultiplexer based on ultralow-refractive-index metamaterial technology. AB - We numerically demonstrate the operation of a novel class of wavelength-division demultiplexing circuit based on photonic crystal waveguides that are entirely synthesized by ultralow-refractive-index metallic nanopillars. The operational principle of the newly proposed device is based on the phenomenon of total external reflections in ultralow-refractive-index metallic photonic crystal structures (metamaterials). In addition we provide detailed design guidelines for optimum device performance. The low propagation losses and compact size, as well as temperature-insensitive operation over a wide temperature range, are only a few of the advantages of the proposed technology, making this new type of demultiplexer an excellent candidate for applications in the visible spectrum. PMID- 16252759 TI - Integrated-optic variable delay line and its application to a low-coherence reflectometer. AB - We propose a large-scale variable delay line based on planar light-wave circuit technology and its application as a reference arm in an optical low-coherence reflectometer. This variable delay line is composed of 16 asymmetrical delay arm pairs sandwiched between 2 optical switches, which select the path for a needed delay. This configuration enables us to eliminate the need for a moving part in the reflectometer. We can scan the reference arm over a length of 262.1 mm with a step of less than 1.0 microm in air and achieve reflectometer sensitivity of about -47 dB. PMID- 16252757 TI - Micrometer-scale all-optical wavelength converter on silicon. AB - We demonstrate a highly integrated micrometer-scale low-power wavelength converter based on the free-carrier dispersion effect in silicon. The conversion is achieved through all-optical modulation of a silicon ring resonator by use of modulated cw control light. The ring resonator has a radius of 5 microm and a Q of approximately 10,000. Both inverted and noninverted modulation are achieved at a bit rate of 0.9 Gbits/s with a control power of 4.5 mW. The scaling of the required control power for operation with respect to the characteristics of the ring resonator is established. PMID- 16252760 TI - Ultralow-noise mode-locked optical pulse trains from an external cavity laser based on a slab coupled optical waveguide amplifier (SCOWA). AB - We report the generation of optical pulse trains with 8.5 fs timing jitter (10 Hz to 10 MHz) from a mode-locked semiconductor laser, with a slab coupled optical waveguide amplifier used as the gain element. This is, to our knowledge, the lowest residual timing jitter reported to date from an actively mode-locked laser. PMID- 16252761 TI - Single-frequency laser oscillator with watts-level output power at 1.5 microm by use of a twisted-mode technique. AB - We report an all-fiber laser oscillator producing as much as 1.9 W of single frequency direct output at 1.5 microm. Spatial gain hole burning in the active fiber has been eliminated by use of a twisted-mode cavity approach. The two short pieces of a polarization-maintaining fiber that were spliced to the ends of the active fiber served as ultracompact quarter-wave plates. To our knowledge, the use of such a wave plate to manipulate the polarization state of light inside a fiber laser cavity is reported here for the first time. The laser output is linearly polarized and delivered through a polarization-maintaining fiber pigtail. We believe that the output power of our laser is the highest among all single-frequency fiber laser oscillators reported to date. PMID- 16252762 TI - Generation of watt-level single-longitudinal-mode output from cladding-pumped short fiber lasers. AB - We generate as much as 1.6 W of continuous-wave 1550 nm single-longitudinal-mode output from a cladding pumped Er-Yb codoped phosphate fiber laser. This power is to our knowledge among the highest in single-longitudinal-mode fiber lasers. The narrowband fiber Bragg grating output coupler is demonstrated to be an effective element for providing the single-longitudinal-mode selection. PMID- 16252763 TI - Mode cycling in microring optical resonators. AB - Electromagnetic resonators are important not only as realizable models of fundamental concepts in classical and quantum physics, based on the existence and properties of eigenmodes, but also in the practical design of lasers, amplifiers, sensors, filters, and delay lines. Coupled-eigenmode systems may be realized via the multiple eigenmodes of a single resonator or by the coupling of a mode across multiple resonators. Mode cycling is demonstrated as a distinct concept of sequential population transfer in coupled multiple-eigenvalue resonators. Based on this principle, a coupled polymeric microring resonator interferometer is fabricated and characterized; the device achieves greater than 30 dB extinction and (loaded) Q approximately 5.5 x 10(3). PMID- 16252764 TI - 131 W 220 fs fiber laser system. AB - We report on an ytterbium-doped photonic-crystal-fiber-based chirped-pulse amplification system delivering 131 W average power 220 fs pulses at 1040 nm center wavelength in a diffraction-limited beam. The pulse repetition rate is 73 MHz, corresponding to a pulse energy of 1.8 microJ and a peak power as high as 8.2 MW. PMID- 16252765 TI - Curvature sensor for the measurement of the static corneal topography and the dynamic tear film topography in the human eye. AB - A system to measure the topography of the first optical surface of the human eye noninvasively by using a curvature sensor is described. The static corneal topography and the dynamic topography of the tear film can both be measured, and the topographies obtained are presented. The system makes possible the study of the dynamic aberrations introduced by the tear film to determine their contribution to the overall ocular aberrations in healthy eyes, eyes with corneal pathologies, and eyes wearing contact lenses. PMID- 16252767 TI - Integrated-grating-induced control of second-harmonic beams in frequency-doubling crystals. AB - We report a novel type of integrated nonlinear photonic device for controlling the generation of several second-harmonic beams. Two-dimensional diffraction gratings are recorded with femtosecond laser pulses at the entrance surface of a frequency-doubling crystal. This periodic spatial modulation of the material surface induces noncollinear propagation of the fundamental input signal in the crystal. By slightly changing the angle of incidence of the seed beam, collinear and noncollinear phase matching can be achieved between different diffraction orders, in this way allowing the efficient generation of several second-harmonic beams. PMID- 16252766 TI - Spectral- and frequency-encoded fluorescence imaging. AB - A method for obtaining fluorescence images with a high number of resolvable points by using spectral and frequency encoding is presented. Broadband excitation light is encoded with a wavelength-dependent frequency modulation and dispersed onto the sample with a grating to simultaneously illuminate an entire image line. The Fourier transform of the frequency-encoded fluorescence emission provides one line of the image. Mechanical scanning along a direction orthogonal to the wavelength-encoded axis allows creation of the two-dimensional fluorescent image. This method is applicable for developing submillimeter diameter endoscopes. The principles of the technique are validated by imaging indocyanine green fluorescence in microfluidic channels. PMID- 16252768 TI - Investigation of laser annealing parameters for optimal laser-damage performance in deuterated potassium dihydrogen phosphate. AB - Laser annealing via preexposure to laser pulses at sub-damage-threshold fluences is known to improve the resistance of KDP crystals to laser-induced damage. Using a specific damage-testing method, we investigate the laser annealing process as a function of fluence and number of preexposure pulses (at 355 nm, 2.5 ns). Our aim is to reveal the key laser parameters in order to devise a practical and efficient protocol for optimizing performance of the material for operation in laser systems in the near UV. Results suggest that a near twofold improvement to the laser-damage performance can be achieved with a limited number of preexposure pulses. PMID- 16252769 TI - Red picosecond pulses generated by frequency doubling a Raman amplified widely tunable 1.3 microm fiber ring laser. AB - We generated 0.66 microm picosecond pulses by second-harmonic generation of the Raman amplified output of a 1.3 microm actively mode-locked fiber ring laser in a periodically poled potassium titanyl phosphate (PPKTP) waveguide. The ring laser produced 9 ps pulses at a 20 GHz repetition frequency, was tunable over 1284-1330 nm, and was based on a semiconductor optical amplifier and a Mach-Zehnder amplitude modulator. The Raman amplifier served both to amplify the ring laser and to compress the pulses as solitons. The spectral flexibility of the amplifiers and the modulator should enable similar configurations to be made at other wavelengths and facilitate efficient frequency doubling in waveguides to other visible wavelengths. PMID- 16252770 TI - Self-processing solgel material for one-step fabrication of micrometer-period sinusoidal phase gratings using the Lloyd's mirror scheme. AB - A novel self-processing silica-zirconia hybrid solgel material has been developed and employed in one-step fabrication of micrometer-period sinusoidal phase gratings. In the process, the gratings with a sinusoidal profile were corrugated on the surface of the solgel film by UV exposure using the Lloyd's mirror setup. No further development or etching step is needed to reveal the sinusoidal profile because the corrugation is formed due to the self-processing property of the solgel material, which is robust enough to be used as an end product. The period, amplitude, diffraction efficiency of the +/-1 order, and surface roughness of one of the fabricated gratings are 0.99 microm, 330 nm, 30.56%, and 1.27 nm, respectively. The new self-processing material is practical and promising for fabrication of micro-optical elements. PMID- 16252771 TI - Tunable photon lifetime in photonic molecules: a concept for delaying an optical signal. AB - We experimentally and theoretically studied the photon lifetime spectral distribution in two coherently coupled spherical microcavities of 3-16 microm diameter forming a photonic molecule, which shows a multipeak narrowband modal structure resulting from lifting of the mode degeneracy with respect to the azimuthal quantum number. The results demonstrate the feasibility of photonic molecules as a basis for a multichannel, wavelength-tunable optical delay-line device, which can be used for delay times in the range 10 ps to 1 ns. PMID- 16252772 TI - Adiabatically tunable optical delay lines and their performance limitations. AB - A simple tunable optical delay line based on adiabatic zone folding in coupled resonator lines is proposed, and its performance is analyzed. The results, valid for all adiabatically tunable optical buffers based on resonators, indicate that, while at low to medium bit rates the performance is improved substantially compared with that of the fixed delay lines, this improvement vanishes for high bit rates and large storage capacities. PMID- 16252773 TI - Resonant tunneling in frustrated total internal reflection. AB - Anomalous light transmission and resonant tunneling in frustrated total internal reflection (FTIR) are theoretically predicted to occur at periodically curved interfaces. For a low-contrast index and for grazing incidence, it is shown that FTIR resonant tunneling provides an optical realization of field-induced barrier transparency in quantum tunneling. PMID- 16252774 TI - Synthetic spectra from rough-surface scattering. AB - We present a method for designing a one-dimensional, deterministic, perfectly conducting rough surface that scatters light at a fixed scattering angle with an intensity whose dependence on the frequency of a plane wave incident normally upon it reproduces the infrared spectrum of a known substance within a specified region of frequencies. Such a surface can therefore be used in a correlation spectrometer for the identification of unknown substances. PMID- 16252775 TI - Experimental study of the stochastic nature of the pulsation self-starting process in passive mode locking. AB - We present an experimental study of the probabilistic nature of pulsation self starting in passively mode-locked lasers. It is a Poissonian process that results from a noise-activated switching barrier. The switching rate from cw operation to pulsation when the laser pump level is turned on has an exponential dependence that is inversely proportional to the square of the laser power. PMID- 16252776 TI - Distortionless light pulse delay in quantum-well Bragg structures. AB - We describe a reflection scheme that allows Bragg-spaced semiconductor quantum wells to be used to trap, store, and release light. We study the temporal and spectral distortion of delayed light pulses and show that this geometry allows multibit delays and offers a high degree of distortion compensation. PMID- 16252777 TI - Mode locking a 1550 nm semiconductor disk laser by using a GaInNAs saturable absorber. AB - Passive mode locking of an optically pumped, InP-based, 1550 nm semiconductor disk laser by using a GaInNAs saturable absorber mirror is demonstrated. To reduce material heating and enable high-power operation, a 50 microm thick diamond heat spreader is bonded to the surface of the gain chip. The laser operates at a repetition frequency of 2.97 GHz and emits near-transform-limited 3.2 ps pulses with an average output power of 120 mW. PMID- 16252778 TI - Femtosecond pulse-shape modulation at nanosecond rates. AB - We report high-rate, computer-controlled femtosecond pulse shaping by use of an electro-optical gallium arsenide optical phased-array modulator with 2304 controlled waveguides. It provides fast modulation speed of both spectral phases and amplitudes. Limited by the driving electronics of our current setup, we were able to update a pulse shape in approximately 30 ns. This technique paves the way toward individual shaping of every single pulse of typical femtosecond mode locked oscillators. PMID- 16252779 TI - In situ characterization of semiconductor saturable absorber mirrors in an operating Yb:KGW mode-locked laser. AB - We report in situ characterization of a semiconductor saturable absorber mirror (SESAM) in an operating Yb:KGW mode-locked laser. The technique may be described as a pump-probe experiment in which the intracavity beam acts as a pump beam while the output of the same laser is used as a test beam for the SESAM reflectivity. At zero delay, the probe pulse overlaps in time with the subsequent intracavity pulse. The method is an alternative to standard pump-probe measurements in situations where the intracavity parameters such as energy fluence onto the SESAM, pulse length, and center wavelength cannot be achieved simultaneously with available lasers. PMID- 16252780 TI - Terahertz polarization imaging. AB - We present a new method to measure the polarization state of a terahertz pulse by using a modified electro-optic sampling setup. To illustrate the power of this method, we show two examples in which the knowledge of the polarization of the terahertz pulse is essential for interpreting the results: spectroscopy measurements on polystyrene foam and terahertz images of a plastic coin. Both measurements show a sample-induced rotation of the terahertz electric field vector, which is surprisingly large and is a strong function of frequency. A promising aspect of our setup is the possibility of simultaneously measuring both transversal electric field components. PMID- 16252781 TI - Generation of single-cycle THz transients with high electric-field amplitudes. AB - Single-cycle terahertz (THz) transients in the frequency range 0.3-7 THz with electric-field amplitudes of more than 400 kV/cm are generated by four-wave mixing of the fundamental and the second harmonic of 25 fs pulses from a Ti:sapphire amplifier in ionized air. These transients are fully characterized by electro-optic sampling with ZnTe and GaP crystals. One can tune the center frequency of the THz transients by varying the length of the incident pulse. The electric-field amplitude increases linearly with the incident pulse energy. PMID- 16252782 TI - Mode-locked laser diode with an ultrafast integrated uni-traveling carrier saturable absorber. AB - A novel two-section integrated mode-locked laser diode (MLLD) with a separate ultrafast uni-traveling carrier (UTC) saturable absorber section and semiconductor optical amplifier gain section is demonstrated. The UTC absorber is composed of a thin p-InGaAsP absorbing layer and an intrinsic InGaAsP collecting layer. By confining the photoexcitation process to the thin highly doped absorbing layer, the diffusion-limited hole extraction process is greatly enhanced. The investigated MLLD produces 600 fs uncompressed optical pulses at a 42 GHz repetition rate. PMID- 16252783 TI - The South Carolina Nurses Association unveils preliminary findings of the nursing school survey on competencies for entry-level registered nurses related to mass casualty incidents. PMID- 16252784 TI - Nursing summit a success, and a great beginning. PMID- 16252785 TI - Addressing the public health nursing shortage: university-health department collaborative experience for BSN students. PMID- 16252788 TI - Psychiatry and disaster: paradoxical relationship. PMID- 16252786 TI - The University of South Carolina Center for Public Health Preparedness: partnering to enhance nursing capacity and capability. PMID- 16252789 TI - Apocalyptic suicide: from a pathological to an eschatological interpretation. AB - BACKGROUND: The authors examine the possible interpretations of apocalyptic suicide--a form of suicide amongst a group which believes that the world will end imminently. MATERIAL: Through a literature search the authors identify a number of paradigmatic cases exemplifying this phenomenon including Jonestown, Waco, The Order of the Solar Temple and Heaven's Gate. DISCUSSION: Interpretations of apocalyptic suicide might consider individual factors--the most common approach- but also the social and eschatological themes which are found in the group's ideological work. Religious violence (including suicide) may be precipitated in a dialectical interaction between group members and the perceived outside world. We consider here the ideological aspects that commonly lead to a dualist epistemology for the group. CONCLUSION: Apocalyptic suicide involves a shift of personal self and agency to the social self of the charismatic group. PMID- 16252790 TI - Subjective experience of recovery from schizophrenia-related disorders and atypical antipsychotics. AB - AIMS: This article investigates the subjective experience of the process of improvement and recovery from the point of view of persons diagnosed (according to research diagnostic criteria) with schizophrenia and schizo-affective disorders. METHODS: A community study of persons using psychiatric services was conducted for a sample of ninety subjects taking atypical antipsychotic medications. Sociodemographic data and clinical ratings were collected to complement the qualitatively developed Subjective Experience of Medication Interview (SEMI), which elicits narrative data on everyday activities, medication and treatment, management of symptoms, expectations concerning recovery, stigma, and quality of life. RESULTS: Recovery was observed through: (1) relatively low ratings of psychiatrically observed symptomatology through BPRS scores; (2) the subjective sense among the majority (77.4%) of participants that taking medication plays a critical role in managing symptoms and avoiding hospitalization; and (3) the subjective sense articulated by the vast majority (80%) that they would recover from their illness and that the quality of their lives would improve (70.6%). CONCLUSION: The overall quality of improvement and recovery is best characterized as an incremental, yet definitively discernable, subjective process. PMID- 16252791 TI - The troubled relationship between psychiatry and sociology. AB - The alienated relationship between psychiatry and sociology is explored. The two disciplines largely took divergent paths after 1970. On the one side, psychiatry manifested a pre-occupation with methodological questions and sought greater medical respectability, with a biomedical approach returning to the fore. Social psychiatry and its underpinning biopsychosocial model became increasingly marginalised and weakened. On the other side, many sociologists turned away from psychiatry and the epidemiological study of mental health problems and increasingly restricted their interest to social theory and qualitative research. An interdisciplinary void ensued, to the detriment of the investigation of social aspects of mental health. PMID- 16252792 TI - Prevalence and correlates of misconduct among ethnically diverse adolescents of Native Hawaiian/ part-Hawaiian and non-Hawaiian ancestry. AB - BACKGROUND: Research on the prevalence and correlates of adolescent misconduct, arrests, and juvenile delinquency has been greatly neglected for the ethnically diverse adolescent groups in Hawai'i (i.e. Asian Americans, Pacific Islanders). AIMS: The aims of the present study are three-fold: (1) to determine whether there are differential rates of adolescent misconduct based on ethnicity, gender, and socioeconomic (SES) factors; (2) on an exploratory basis, to ascertain the demographic, social-cultural, psychological-psychiatric, and school-related correlates of misconduct, with some focus on cultural identification; and (3) to derive a parsimonious model of the correlates for an ethnically diverse group of adolescents by disentangling cultural identification from other variables including ethnicity. METHODS: Data were analyzed from the Native Hawaiian Mental Health Research Development Program, consisting of 2,732 Asian/Pacific Islander youths. Misconduct was operationally defined as 'was arrested or got in serious trouble with the law' within the past six months via adolescent self-report. Of added importance were the inclusion of a valid measure of Hawaiian cultural identification and actual school data. RESULTS: Significantly higher rates were obtained for Native Hawaiians, males, and adolescents whose main wage earners' educational attainment was at the high school level or less. Absences may be a more important marker for females than males in the prediction of arrests and serious trouble with the law. The most parsimonious model of self-reported arrests/serious trouble with the law consisted of four variables in the following order of importance: substance use, actual grade-point average, aggressive symptoms, and gender. CONCLUSION: This is one of the first major studies to examine the prevalence and risk-protective factors of misconduct for adolescents of Asian/Pacific Islander ancestry, and the first to include a valid measure of cultural identification and actual school data. The findings indicated that gender-specific prevention and intervention programs are needed. However, the issues are also complex whereby the use of substances, school performance, and aggression must be taken into account in deriving successful programs. The lack of sustained association between culturally based factors (e.g., ethnicity, Hawaiian cultural identification) and misconduct in the final parsimonious model suggested that the culturally based variables are not direct causes of misconduct, but rather correlates of misconduct that are over-shadowed by more substantive factors, such as substance use and academic performance, the latter being variables that must be addressed for youths across the United States. The present results reaffirmed the common correlates of misconduct across different ethnic groups. Further research is needed in the causal relationships among these important associations with misconduct. PMID- 16252793 TI - Family burden during exacerbation of schizophrenia: quantification and determinants of additional costs. AB - BACKGROUND/AIMS: Family members of people with schizophrenia are heavily burdened. Thus, we aimed at quantifying the amount of additional efforts during the two weeks before hospitalisation of acute exacerbating psychosis compared to the time before. METHODS: By means of the semi-structured 'Interview for Measuring the Burden on the Family' we interviewed relatives of people with schizophrenia. To quantify additional burden we asked how much time a person who would take over the additional burden has to afford. Subscales of different sources of burden and total scales were correlated. RESULTS: An additional 8.9 hours per day would have to be spent in favour of the patient, which results in additional costs of Euro 3,115-Euro 4,984 (ca. pounds 2,100-3,400) for two weeks. The main sources of additional burden are quantity, quality and nature of contacts to the affected. This includes the frequency of contact to the affected, the whereabouts and symptoms of the affected, as well as the social contacts of the relatives. CONCLUSIONS: Our results clarify the quantitative aspect of care provided by relatives of people with schizophrenia. Apart from the practical and emotional help to relatives the development of a diversified network of community psychiatry services, e.g. day and night hospitals, as well as crisis accommodations should be established. PMID- 16252794 TI - The Social Functioning Questionnaire: a rapid and robust measure of perceived functioning. AB - BACKGROUND: The Social Functioning Questionnaire (SFQ), an eight-item self-report scale (score range 0-24), was developed from the Social Functioning Schedule (SFS), a semi-structured interview which has been used primarily with non psychotic patients and has good test-retest and inter-rater reliability as well as construct validity. The SFQ was developed following the need for a quick assessment of perceived social function. AIMS: To give further details of old and new data sets from studies involving over 4000 subjects assessed with the SFQ illustrating its epidemiological and clinical associations. METHOD: New data were analysed from a national epidemiological study, a comparison of key-worker and subject versions of the SFQ, and reanalysis of data from three earlier clinical studies, of psychiatric emergencies, general practice psychiatric patients and those with recurrent psychotic illnesses. These data were examined further to determine their range, their relationship to other clinical measures, and change over time in clinical trials. RESULTS: The population mean score in 4164 subjects was 4.6 and the data from all studies suggested that a score of 10 or more indicated poor social functioning. Those presenting as psychiatric emergencies had the poorest social function (mean 11.4) and psychiatric patients from general practice the best function (mean 7.7) of the clinical populations. The eight item scores had a normal distribution in psychiatric populations and a skewed one in a normal population; scores were relatively stable over the short (weeks) and long term (months), and were high in the presence of acute mental health disturbance and personality disorder, giving support to the validity of the scale. The results from a UK sample of a randomly selected population specifically weighted for ethnic minorities showed similar social function across groups. PMID- 16252795 TI - The stigma of mental illness in Germany: a trend analysis. AB - BACKGROUND: While there are some indications that the mental health literacy of the public has improved within recent years, the findings concerning its attitudes towards the mentally ill are quite inconsistent. AIMS: The aim of this study is to examine whether any changes have taken place in Germany over the last decade regarding the stigmatisation of mentally ill people. METHOD: In 2001, a representative population survey was carried out among the German adult population, using the same sampling procedure and the same measure for the assessment of perceived stigma as in a previous survey that had been conducted in 1990. RESULTS: Our findings suggest that in 2001, the German public was somewhat less inclined to believe that former mental patients are exposed to stigmatisation than a decade ago. While this holds especially true for the devaluation of patients, the picture is more mixed for discrimination. The trend towards less perceived stigmatisation is particularly pronounced among people who are familiar with psychiatric treatment. CONCLUSIONS: Since a substantial amount of perceived stigma still persists, further efforts to reduce the stigma attached to mental illness are needed. PMID- 16252796 TI - Taoism, mental health and the limits of language. PMID- 16252797 TI - Mental health promotion in forensic & correctional environments. AB - I hope this brief glimpse into the concept of mental health promotion within forensic and correctional environments will challenge nurses to explore creative ways in which a mental health agenda can be formulated and actualized in practice. PMID- 16252798 TI - Fostering hope in incarcerated older adults. PMID- 16252799 TI - Sexually motivated child abductors: forensic evaluation. AB - 1. Forensic nursing evaluations of perpetrators require (at minimum) information on the crime, crime scene photographs (if possible), victim statements, official records, and perpetrator interviews. 2. If multiple crime scenes exist, each needs to be detailed for an evaluation report. 3. Sexually motivated crimes are usually committed based on a well-defined fantasy. PMID- 16252800 TI - Client-centered care for individuals with dual diagnoses in the justice system. AB - 1. The characteristics of people with developmental disabilities, such as poor judgment, low frustration tolerance, lack of impulse control, and naivete, compounded by the symptoms of a mental illness, contribute to their susceptibility to engage in problematic or criminal behaviors. 2. Nurses should recognize that conducting risk assessments for this population may take time and could involve modifying risk assessments to the individuals' abilities. 3. Providing appropriate care for this vulnerable population requires a long-term, interdisciplinary approach and quality community support services PMID- 16252801 TI - ADHD in juvenile offenders: treatment issues nurses need to know. AB - 1. When working with juvenile offenders with ADHD, advanced practice nurses should consider prescribing stimulant medication in conjunction with other psychosocial interventions to achieve maximum treatment success. 2. Nurses working with young people with ADHD in the justice system must carefully coordinate care among all health care providers and juvenile justice personnel, including attorneys, probation/parole officers, and court assessment personnel. 3. Advocating for the individual needs of clients and their families is critical; often, this includes advocating for care that is not only evidence based, but also culturally competent. PMID- 16252802 TI - Total antioxidant status in the blood serum of rats exposed to N-nitroso compounds and nitric oxide synthase inhibitors. AB - In this study, the total antioxidant status (TAS) was assayed in the blood serum of rats pretreated per os with either N-nitrosodiethylamine (NDEA) (0.1 mg/kg b.w./day) or N-methyl-N-nitrosourea (NMU) (0.1 mg/kg b.w./day) for 30 days. The animals were also dosed per os with spermidine (SPR) (10 mg/kg b.w./day) for a first 21 day period, and Nomega)-nitro-L-arginine methyl ester (L-NAME) (10 mg/kg b.w./day) given to animals for 3 days (days 22-24), respectively. Nitric oxide synthase (NOS) inhibitor, L-NAME was found to mitigate TAS levels in the blood serum of rats pretreated with NDEA and NMU. No such changes were found in animals dosed with L-NAME only nor even with L-NAME and spermidine, respectively. Since spermidine, also known as an inhibitor of iNOS synthesis, elevated TAS levels in rats dosed with L-NAME and NDEA/NMU, the polyamine was suggested to modify the NOS/NO origin to serve the physiological level of the total anti-oxidant status in rat blood serum. PMID- 16252803 TI - [Evaluation of the usefulness of SIS and MS/MS mode for the determination of persistent organochlorine compounds and polychlorinated biphenyls in human specimens]. AB - This study aimed of examining of the applicability of the SIS (Selected Ion Storage) and the MS/ MS (Tandem Mass Spectrometry) methods for analyzing organochlorine pesticide (OCs) residues and PCBs in human specimens. The samples of human abdominal adipose tissue, women's breast adipose tissue, and human milk were used as study material. The samples were analyzed using the gas chromatography method with EC and MS detectors. MS/MS and SIS mode allow to eliminate system-based (e.g., column, septum) ions and the biological samples extract impurities' ions from the background to achieve the analytical system sensitive enough for trace analysis of organochlorine compounds in human specimens. PMID- 16252804 TI - [Selected seafoods as the components of a highly nutritional diet]. AB - Shellfish products available on the Polish market were analyzed for the content of protein, fat, carbohydrates, water, ash, energy value and vitamins B. Nutrition value of the analyzed products was determined by analytical methods applying in food chemistry. Concentrations of niacin and vitamin B6 were determined by microbiological methods. The concentration of the selected shellfish in 100 g of the analyzed products and percentage of realization on recommended level of nutritive components by analyzed products for adults were determined. PMID- 16252805 TI - [Popularity of "fast-food" products consumption by school youth in urban and country secondary schools from Mazowsze area]. AB - The aim of this study was the evaluation of frequency intake of "fast-food" products by young people in urban and country secondary schools. The group of 282 pupils (14 years) was investigated. Frequency intake was estimated using questionnaire. In the populations of boys in town frequency of consuming "fast food" products has risen in comparison with boys from country. Three groups comprised adolescent girls (for a change) have not difference in frequency of "fast-food" consumption. PMID- 16252806 TI - [Chosen dietary habits in a cohort of students of the Medical University of Bialystok in the years 2000-2003]. AB - The aim of the study was to estimate chosen dietary habits of a cohort of students in the years 2000-2003. The frequency of the intake of chosen groups of food products, the number of meals and snacks between meals were considered. The highest percentage of both male and female students involved in the questionnaire investigations in the study period consumed three meals a day. The number of students having snacks between meals (every third person) did not change during that period. However, the decrease was observed in drinking by students one glass of milk a day. In the years 2000-2003, the reduction in the frequency of consumption of whole-grain breads and cereals in female students and its increase in male students was noted. The number of men consuming meat and its products was increased. More than half of the students of both sexes consumed fish once a week and this tendency was maintained for the whole study period. An increase was noted in the percentage of female students consuming vegetable salads and in the percentage of male students eating fruit. PMID- 16252807 TI - [Hair vanadium content and nutritional status of students of the Medical University of Bialystok]. AB - The aim of the study was to determine the vanadium content in the hair of students of the Medical University of Bialystok and to find out whether its level in the organism measured basing on its hair content correlates with the nutritional status. The study involved 134, 127 and 125 students in the years 2000-2003, respectively. Hair vanadium content was determined using the atomic absorption spectrophotometry method with atomization in a graphite cuvette (ET AAS) on a Hitachi Z-5000 apparatus. The nutritional status was evaluated based on the body mass index BMI. Higher vanadium content was detected in the hair of female students compared to male students. Negative correlation was noted between vanadium content in the hair of female students and their nutritional status. PMID- 16252808 TI - [Nutritional behavior of pre-school children]. AB - The work presents the analysis of nutritional behavior of pre-school children. The most likely food for children are fast-food, because their like ate in the restaurant, where parents accepted their choice. The parents and the grand parents entertain the sweets than recompense for their good behavior. The parents very often, make mistake thinking that this what is good for them, is good also for their children. PMID- 16252809 TI - [Concentration of several bioelements in bread and other plant supplements]. AB - Concentrations of Ca, P, Fe, Zn and Cu were determined in grain products such as bread and crispbread with natural supplements, and also in seeds and other cereal products available on Polish market, added to improve bread's quality and nutritional value. The contents of minerals in bread were as follow: 35.3-78.4 mg Ca; 88.4-364 mg P; 3.03-5.63 mg Fe; 1.96-3.15 mg Zn and 0.27-0.64 mg Cu in 100 g of product. Among the analyzed products, the highest level of calcium was recorded for poppy and sesame seeds. Based on the data obtained it was possible to estimate the realization of the recommended daily intake of bioelements with the analyzed products for an adult person. PMID- 16252810 TI - [Iron and manganese in selected herbs and herbal-fruit teas]. AB - The aim of this study was the determination iron and manganese contents in selected herbs, herbal-fruit teas and their water extracts. The samples were mineralized at temperature of 450 degrees C. The iron and manganese contents were determined by atomic absorption spectrometry (AAS) directly from the mineralizates using SOLAAR M5 appliance produced by a US-based Thermo Elemental. The iron contents in dry herbs ranged, at average, from 97.66 to 348.65 microg/g and from 48.95 to 118.40 microg/g in herbal-fruit teas, the manganese contents from 43.25 to 604.80 microg/g in herbs and from 106.75 to 197.00 microg/g in herbal-fruit teas. The iron and manganese contents in dry herbs were comparable to results obtained by authors of the other papers. Infusions and decoctions of the examined herbsand herbal-fruit teas contain the amounts of these elements that do not harm the health and may constitute an additional source of iron and manganese for the human organism. PMID- 16252811 TI - [Assessment of zinc and copper contents in the hair as the test of environmental pollution of Gliwice]. AB - This work presents the zinc and copper content in the hair measured by the flame atomic absorption spectrometric (FAAS). Prior to the FAAS analysis mineralized samples in HNO3+H2O2 (2:1 v/v) solution using microwave technique. Early, all samples washed with water-acetone-water, because grease and dust on the surface of hair may contribute of the element concentrations of hair. Hair samples were collected from people who were not exposed to the contact with copper or zinc. The influence of the variable place of residence, age, sex, was also examined. The mean content of copper was 11.51 +/- 1.27 mg/g dry weight and zinc 146.32 +/- 14.93 mg/g dry weight. The places of residence sex and age were variables that significantly affected copper and zinc concentrations. Men's had more copper and zinc in their hair than women's, whereas hair zinc and copper levels were significantly correlated with the place of the residence and age. The methods were free from interferences, reliable and reproducible. For all metals present in real samples, precision was better than 0.4% (RSD). PMID- 16252813 TI - Load characterization and anomaly detection for voice over IP traffic. AB - We consider the problem of traffic anomaly detection in IP networks. Traffic anomalies typically arise when there is focused overload or when a network element fails and it is desired to infer these purely from the measured traffic. We derive new general formulae for the variance of the cumulative traffic over a fixed time interval and show how the derived analytical expression simplifies for the case of voice over IP traffic, the focus of this paper. To detect load anomalies, we show it is sufficient to consider cumulative traffic over relatively long intervals such as 5 min. We also propose simple anomaly detection tests including detection of over/underload. This approach substantially extends the current practice in IP network management where only the first-order statistics and fixed thresholds are used to identify abnormal behavior. We conclude with the application of the scheme to field data from an operational network. PMID- 16252812 TI - [Microbiological quality of indoor air at the School of Building and Environmental Engineering at Bialystok University of Technology]. AB - The investigation of microbiological rate of indoor air pollution on Faculty of Building and Environmental Engineering at Bialystok University of Technology were made by sedimentation method in accordance with Polish standards (PN-89/Z 04111/01,02,03). Six series of measurements were carried out from autumn 2002 to spring 2003. The results show bad microbiological quality of indoor air on Faculty of Building and Environmental Engineering at Bialystok University of Technology. It was found that the number of Staphylococcus, Actinomycetales as well as the total count of bacteria were too high and broke the Polish regulations of the clear air. Because of the students' and other workers' safety, monitoring of microbiological pollution of the indoor air must be done and existing emergency to improve the quality of the air must be lead. PMID- 16252814 TI - Detecting application-level failures in component-based internet services. AB - Most Internet services (e-commerce, search engines, etc.) suffer faults. Quickly detecting these faults can be the largest bottleneck in improving availability of the system. We present Pinpoint, a methodology for automating fault detection in Internet services by: 1) observing low-level internal structural behaviors of the service; 2) modeling the majority behavior of the system as correct; and 3) detecting anomalies in these behaviors as possible symptoms of failures. Without requiring any a priori application-specific information, Pinpoint correctly detected 89%-96% of major failures in our experiments, as compared with 20%-70% detected by current application-generic techniques. PMID- 16252815 TI - Probabilistic approaches to fault detection in networked discrete event systems. AB - In this paper, we consider distributed systems that can be modeled as finite state machines with known behavior under fault-free conditions, and we study the detection of a general class of faults that manifest themselves as permanent changes in the next-state transition functionality of the system. This scenario could arise in a variety of situations encountered in communication networks, including faults occurred due to design or implementation errors during the execution of communication protocols. In our approach, fault diagnosis is performed by an external observer/diagnoser that functions as a finite state machine and which has access to the input sequence applied to the system but has only limited access to the system state or output. In particular, we assume that the observer/diagnoser is only able to obtain partial information regarding the state of the given system at intermittent time intervals that are determined by certain synchronizing conditions between the system and the observer/diagnoser. By adopting a probabilistic framework, we analyze ways to optimally choose these synchronizing conditions and develop adaptive strategies that achieve a low probability of aliasing, i.e., a low probability that the external observer/diagnoser incorrectly declares the system as fault-free. An application of these ideas in the context of protocol testing/classification is provided as an example. PMID- 16252816 TI - Statistical analysis of network traffic for adaptive faults detection. AB - This paper addresses the problem of normal operation baselining for automatic detection of network anomalies. A model of network traffic is presented in which studied variables are viewed as sampled from a finite mixture model. Based on the stochastic approximation of the maximum likelihood function, we propose baselining network normal operation, using the asymptotic distribution of the difference between successive estimates of model parameters. The baseline random variable is shown to be stationary, with mean zero under normal operation. Anomalous events are shown to induce an abrupt jump in the mean. Detection is formulated as an online change point problem, where the task is to process the baseline random variable realizations, sequentially, and raise alarms as soon as anomalies occur. An analytical expression of false alarm rate allows us to choose the design threshold, automatically. Extensive experimental results on a real network showed that our monitoring agent is able to detect unusual changes in the characteristics of network traffic, adapt to diurnal traffic patterns, while maintaining a low alarm rate. Despite large fluctuations in network traffic, this work proves that tailoring traffic modeling to specific goals can be efficiently achieved. PMID- 16252817 TI - Condition monitoring of 3G cellular networks through competitive neural models. AB - We develop an unsupervised approach to condition monitoring of cellular networks using competitive neural algorithms. Training is carried out with state vectors representing the normal functioning of a simulated CDMA2000 network. Once training is completed, global and local normality profiles (NPs) are built from the distribution of quantization errors of the training state vectors and their components, respectively. The global NP is used to evaluate the overall condition of the cellular system. If abnormal behavior is detected, local NPs are used in a component-wise fashion to find abnormal state variables. Anomaly detection tests are performed via percentile-based confidence intervals computed over the global and local NPs. We compared the performance of four competitive algorithms [winner take-all (WTA), frequency-sensitive competitive learning (FSCL), self-organizing map (SOM), and neural-gas algorithm (NGA)] and the results suggest that the joint use of global and local NPs is more efficient and more robust than current single threshold methods. PMID- 16252818 TI - An artificial immune system approach with secondary response for misbehavior detection in mobile ad hoc networks. AB - In mobile ad hoc networks, nodes act both as terminals and information relays, and they participate in a common routing protocol, such as dynamic source routing (DSR). The network is vulnerable to routing misbehavior, due to faulty or malicious nodes. Misbehavior detection systems aim at removing this vulnerability. In this paper, we investigate the use of an artificial immune system (AIS) to detect node misbehavior in a mobile ad hoc network using DSR. The system is inspired by the natural immune system (IS) of vertebrates. Our goal is to build a system that, like its natural counterpart, automatically learns, and detects new misbehavior. We describe our solution for the classification task of the AIS; it employs negative selection and clonal selection, the algorithms for learning and adaptation used by the natural IS. We define how we map the natural IS concepts such as self, antigen, and antibody to a mobile ad hoc network and give the resulting algorithm for classifying nodes as misbehaving. We implemented the system in the network simulator Glomosim; we present detection results and discuss how the system parameters affect the performance of primary and secondary response. Further steps will extend the design by using an analogy to the innate system, danger signal, and memory cells. PMID- 16252819 TI - Adaptive diagnosis in distributed systems. AB - Real-time problem diagnosis in large distributed computer systems and networks is a challenging task that requires fast and accurate inferences from potentially huge data volumes. In this paper, we propose a cost-efficient, adaptive diagnostic technique called active probing. Probes are end-to-end test transactions that collect information about the performance of a distributed system. Active probing uses probabilistic reasoning techniques combined with information-theoretic approach, and allows a fast online inference about the current system state via active selection of only a small number of most informative tests. We demonstrate empirically that the active probing scheme greatly reduces both the number of probes (from 60% to 75% in most of our real life applications), and the time needed for localizing the problem when compared with nonadaptive (preplanned) probing schemes. We also provide some theoretical results on the complexity of probe selection, and the effect of "noisy" probes on the accuracy of diagnosis. Finally, we discuss how to model the system's dynamics using dynamic Bayesian networks (DBNs), and an efficient approximate approach called sequential multifault; empirical results demonstrate clear advantage of such approaches over "static" techniques that do not handle system's changes. PMID- 16252820 TI - Long-term forecasting of internet backbone traffic. AB - We introduce a methodology to predict when and where link additions/upgrades have to take place in an Internet protocol (IP) backbone network. Using simple network management protocol (SNMP) statistics, collected continuously since 1999, we compute aggregate demand between any two adjacent points of presence (PoPs) and look at its evolution at time scales larger than 1 h. We show that IP backbone traffic exhibits visible long term trends, strong periodicities, and variability at multiple time scales. Our methodology relies on the wavelet multiresolution analysis (MRA) and linear time series models. Using wavelet MRA, we smooth the collected measurements until we identify the overall long-term trend. The fluctuations around the obtained trend are further analyzed at multiple time scales. We show that the largest amount of variability in the original signal is due to its fluctuations at the 12-h time scale. We model inter-PoP aggregate demand as a multiple linear regression model, consisting of the two identified components. We show that this model accounts for 98% of the total energy in the original signal, while explaining 90% of its variance. Weekly approximations of those components can be accurately modeled with low-order autoregressive integrated moving average (ARIMA) models. We show that forecasting the long term trend and the fluctuations of the traffic at the 12-h time scale yields accurate estimates for at least 6 months in the future. PMID- 16252821 TI - A fuzzy-based approach to remove clock skew and reset from one-way delay measurement. AB - One-way delay (OWD) traces are important measurements for analyzing end-to-end performance on the Internet. It is still a great challenge to provide a scalable solution for large-scale OWD measurement. Because the clocks at end systems are usually not synchronized, the OWD measurements are often inaccurate. For the more challenging case with clock resets to some reference times during the measurement, the OWD measurements are more inaccurate. Furthermore, the measurement data often exhibit considerable network-induced noise when the network is heavily loaded. All the existing OWD measurement techniques, such as linear programming and convex-hull approach (CHA), try to solve this problem by deterministic mathematics model. However, they often fail to distinguish clock resets from temporary Internet congestion. Based on the fuzzy-clustering analysis, this paper proposes a new algorithm to estimate and remove the clock skews and resets from measurement results. This algorithm has been implemented as a tool called fuzzy-based OWD corrector (FOC). The paper then presents OWD measurements of several Internet paths using FOC. Numerical experiments demonstrate that FOC is more accurate and robust than the existing techniques. FOCs computation complexity O(N) is similar to that of CHA and its computing time is much less than that of convex-hull technique. PMID- 16252822 TI - Sparse basis selection: new results and application to adaptive prediction of video source traffic. AB - Real-time prediction of video source traffic is an important step in many network management tasks such as dynamic bandwidth allocation and end-to-end quality-of service (QoS) control strategies. In this paper, an adaptive prediction model for MPEG-coded traffic is developed. A novel technology is used, first developed in the signal processing community, called sparse basis selection. It is based on selecting a small subset of inputs (basis) from among a large dictionary of possible inputs. A new sparse basis selection algorithm is developed that is based on efficiently updating the input selection adaptively. When a new measurement is received, the proposed algorithm updates the selected inputs in a recursive manner. Thus, adaptability is not only in the weight adjustment, but also in the dynamic update of the inputs. The algorithm is applied to the problem of single-step-ahead prediction of MPEG-coded video source traffic, and the developed method achieves improved results, as compared to the published results in the literature. The present analysis indicates that the adaptive feature of the developed algorithm seems to add significant overall value. PMID- 16252823 TI - Fuzzy-based adaptive bandwidth control for loss guarantees. AB - This paper presents the use of adaptive bandwidth control (ABC) for a quantitative packet loss rate guarantee to aggregate traffic in packet switched networks. ABC starts with some initial amount of bandwidth allocated to a queue and adjusts it over time based on online measurements of system states to ensure that the allocated bandwidth is just enough to attain the specified loss requirement. Consequently, no a priori detailed traffic information is required, making ABC more suitable for efficient aggregate quality of service (QoS) provisioning. We propose an ABC algorithm called augmented Fuzzy (A-Fuzzy) control, whereby fuzzy logic control is used to keep an average queue length at an appropriate target value, and the measured packet loss rate is used to augment the standard control to achieve better performance. An extensive simulation study based on both theoretical traffic models and real traffic traces under a wide range of system configurations demonstrates that the A-Fuzzy control itself is highly robust, yields high bandwidth utilization, and is indeed a viable alternative and improvement to static bandwidth allocation (SBA) and existing adaptive bandwidth allocation schemes. Additionally, we develop a simple and efficient measurement-based admission control procedure which limits the amount of input traffic in order to maintain the performance of the A-Fuzzy control at an acceptable level. PMID- 16252824 TI - Dynamic neural-based buffer management for Queuing systems with self-similar characteristics. AB - Buffer management in queuing systems plays an important role in addressing the tradeoff between efficiency measured in terms of overall packet loss and fairness measured in terms of individual source packet loss. Complete partitioning (CP) of a buffer with the best fairness characteristic and complete sharing (CS) of a buffer with the best efficiency characteristic are at the opposite ends of the spectrum of buffer management techniques. Dynamic partitioning buffer management techniques aim at addressing the tradeoff between efficiency and fairness. Ease of implementation is the key issue when determining the practicality of a dynamic buffer management technique. In this paper, two novel dynamic buffer management techniques for queuing systems accommodating self-similar traffic patterns are introduced. The techniques take advantage of the adaptive learning power of perceptron neural networks when applied to arriving traffic patterns of queuing systems. Relying on the water-filling approach, our proposed techniques are capable of coping with the tradeoff between packet loss and fairness issues. Computer simulations reveal that both of the proposed techniques enjoy great efficiency and fairness characteristics as well as ease of implementation. PMID- 16252825 TI - Neural and fuzzy computation techniques for playout delay adaptation in VoIP networks. AB - Playout delay adaptation algorithms are often used in real time voice communication over packet-switched networks to counteract the effects of network jitter at the receiver. Whilst the conventional algorithms developed for silence suppressed speech transmission focused on preserving the relative temporal structure of speech frames/packets within a talkspurt (intertalkspurt adaptation), more recently developed algorithms strive to achieve better quality by allowing for playout delay adaptation within a talkspurt (intratalkspurt adaptation). The adaptation algorithms, both intertalkspurt and intratalkspurt based, rely on short term estimations of the characteristics of network delay that would be experienced by up-coming voice packets. The use of novel neural networks and fuzzy systems as estimators of network delay characteristics are presented in this paper. Their performance is analyzed in comparison with a number of traditional techniques for both inter and intratalkspurt adaptation paradigms. The design of a novel fuzzy trend analyzer system (FTAS) for network delay trend analysis and its usage in intratalkspurt playout delay adaptation are presented in greater detail. The performance of the proposed mechanism is analyzed based on measured Internet delays. Index Terms-Fuzzy delay trend analysis, intertalkspurt, intratalkspurt, multilayer perceptrons (MLPs), network delay estimation, playout buffering, playout delay adaptation, time delay neural networks (TDNNs), voice over Internet protocol (VoIP). PMID- 16252826 TI - Neural approximation of open-loop feedback rate control in satellite networks. AB - A resource allocation problem for a satellite network is considered, where variations of fading conditions are added to those of traffic load. Since the capacity of the system is finite and divided in finite discrete portions, the resource allocation problem reveals to be a discrete stochastic programming one, which is typically NP-Hard. In practice, a good approximation of the optimal solution could be obtained through the adoption of a closed-form expression of the performance measure in steady-state conditions. Once we have summarized the drawbacks of such optimization strategy, we address two novel optimization approaches. The first one derives from Gokbayrak and Cassandras and is based on the minimization over the discrete constraint set using an estimate of the gradient, obtained through a "relaxed continuous extension" of the performance measure. The computation of the gradient estimation is based on infinitesimal perturbation analysis (IPA). Neither closed forms of the performance measures, nor additional feedbacks concerning the state of the system and very mild assumptions about the stochastic environment are requested. The second one is the main contribution of the present work, and is based on an open-loop feedback control (OLFC) strategy, aimed at providing optimal reallocation strategies as functions of the state of the network. The optimization approach leads us to a functional optimization problem, and we investigate the adoption of a neural network-based technique, in order to approximate its solution. As is shown in the simulation results, we obtain near-optimal reallocation strategies with a small real time computational effort and avoid the suboptimal transient periods introduced by the IPA gradient descent algorithm. PMID- 16252827 TI - Adaptive power control for wireless networks using multiple controllers and switching. AB - Controlling transmitted power in a wireless network is critical for maintaining quality of service, maximizing channel utilization and minimizing near-far effect for suboptimal receivers. In this paper, a general proportional-integral derivative (PID) type algorithm for controlling transmitted powers in wireless networks is studied and a systematic way to adapt or tune the parameters of the controller in a distributed fashion is suggested. The proposed algorithm utilizes multiple candidate PID gains. Depending on the prevailing channel conditions, it selects an optimal PID gain from the candidate gain set at each instant and places it in the feedback loop. The algorithm is data driven and can distinguish between stabilizing and destabilizing controller gains as well as rank the stabilizing controllers based on their performance. Simulation results indicate that the proposed scheme performs better than several candidate controllers, including a well known distributed power control (DPC) algorithm. PMID- 16252828 TI - A self-learning call admission control scheme for CDMA cellular networks. AB - In the present paper, a call admission control scheme that can learn from the network environment and user behavior is developed for code division multiple access (CDMA) cellular networks that handle both voice and data services. The idea is built upon a novel learning control architecture with only a single module instead of two or three modules in adaptive critic designs (ACDs). The use of adaptive critic approach for call admission control in wireless cellular networks is new. The call admission controller can perform learning in real-time as well as in offline environments and the controller improves its performance as it gains more experience. Another important contribution in the present work is the choice of utility function for the present self-learning control approach which makes the present learning process much more efficient than existing learning control methods. The performance of our algorithm will be shown through computer simulation and compared with existing algorithms. PMID- 16252829 TI - Randomized algorithms for stability and robustness analysis of high-speed communication networks. AB - This paper initiates a study toward developing and applying randomized algorithms for stability of high-speed communication networks. The focus is on congestion and delay-based flow controllers for sources, which are "utility maximizers" for individual users. First, we introduce a nonlinear algorithm for such source flow controllers, which uses as feedback aggregate congestion and delay information from bottleneck nodes of the network, and depends on a number of parameters, among which are link capacities, user preference for utility, and pricing. We then linearize this nonlinear model around its unique equilibrium point and perform a robustness analysis for a special symmetric case with a single bottleneck node. The "symmetry" here captures the scenario when certain utility and pricing parameters are the same across all active users, for which we derive closed-form necessary and sufficient conditions for stability and robustness under parameter variations. In addition, the ranges of values for the utility and pricing parameters for which stability is guaranteed are computed exactly. These results also admit counterparts for the case when the pricing parameters vary across users, but the utility parameter values are still the same. In the general nonsymmetric case, when closed-form derivation is not possible, we construct specific randomized algorithms which provide a probabilistic estimate of the local stability of the network. In particular, we use Monte Carlo as well as quasi-Monte Carlo techniques for the linearized model. The results obtained provide a complete analysis of congestion control algorithms for internet style networks with a single bottleneck node as well as for networks with general random topologies. PMID- 16252830 TI - New MPLS network management techniques based on adaptive learning. AB - The combined use of the differentiated services (DiffServ) and multiprotocol label switching (MPLS) technologies is envisioned to provide guaranteed quality of service (QoS) for multimedia traffic in IP networks, while effectively using network resources. These networks need to be managed adaptively to cope with the changing network conditions and provide satisfactory QoS. An efficient strategy is to map the traffic from different DiffServ classes of service on separate label switched paths (LSPs), which leads to distinct layers of MPLS networks corresponding to each DiffServ class. In this paper, three aspects of the management of such a layered MPLS network are discussed. In particular, an optimal technique for the setup of LSPs, capacity allocation of the LSPs and LSP routing are presented. The presented techniques are based on measurement of the network state to adapt the network configuration to changing traffic conditions. PMID- 16252831 TI - Web content management by self-organization. AB - We present a new method for content management and knowledge discovery using a topology-preserving neural network. The method, termed topological organization of content (TOC), can generate a taxonomy of topics from a set of unannotated, unstructured documents. The TOC consists of a hierarchy of self-organizing growing chains (GCs), each of which can develop independently in terms of size and topics. The dynamic development process is validated continuously using a proposed entropy-based Bayesian information criterion (BIC). Each chain meeting the criterion spans child chains, with reduced vocabularies and increased specializations. This results in a topological tree hierarchy, which can be browsed like a table of contents directory or web portal. A brief review is given on existing methods for document clustering and organization, and clustering validation measures. The proposed approach has been tested and compared with several existing methods on real world web page datasets. The results have clearly demonstrated the advantages and efficiency in content organization of the proposed method in terms of computational cost and representation. The TOC can be easily adapted for large-scale applications. The topology provides a unique, additional feature for retrieving related topics and confining the search space. PMID- 16252832 TI - Self-organizing network services with evolutionary adaptation. AB - This paper proposes a novel framework for developing adaptive and scalable network services. In the proposed framework, a network service is implemented as a group of autonomous agents that interact in the network environment. Agents in the proposed framework are autonomous and capable of simple behaviors (e.g., replication, migration, and death). In this paper, an evolutionary adaptation mechanism is designed using genetic algorithms (GAs) for agents to evolve their behaviors and improve their fitness values (e.g., response time to a service request) to the environment. The proposed framework is evaluated through simulations, and the simulation results demonstrate the ability of autonomous agents to adapt to the network environment. The proposed framework may be suitable for disseminating network services in dynamic and large-scale networks where a large number of data and services need to be replicated, moved, and deleted in a decentralized manner. PMID- 16252833 TI - Back pressure based multicast scheduling for fair bandwidth allocation. AB - We study the fair allocation of bandwidth in multicast networks with multirate capabilities. In multirate transmission, each source encodes its signal in layers. The lowest layer contains the most important information and all receivers of a session should receive it. If a receiver's data path has additional bandwidth, it receives higher layers which leads to a better quality of reception. The bandwidth allocation objective is to distribute the layers fairly. We present a computationally simple, decentralized scheduling policy that attains the maxmin fair rates without using any knowledge of traffic statistics and layer bandwidths. This policy learns the congestion level from the queue lengths at the nodes, and adapts the packet transmissions accordingly. When the network is congested, packets are dropped from the higher layers; therefore, the more important lower layers suffer negligible packet loss. We present analytical and simulation results that guarantee the maxmin fairness of the resulting rate allocation, and upper bound the packet loss rates for different layers. PMID- 16252834 TI - Spatial-temporal modeling of malware propagation in networks. AB - Network security is an important task of network management. One threat to network security is malware (malicious software) propagation. One type of malware is called topological scanning that spreads based on topology information. The focus of this work is on modeling the spread of topological malwares, which is important for understanding their potential damages, and for developing countermeasures to protect the network infrastructure. Our model is motivated by probabilistic graphs, which have been widely investigated in machine learning. We first use a graphical representation to abstract the propagation of malwares that employ different scanning methods. We then use a spatial-temporal random process to describe the statistical dependence of malware propagation in arbitrary topologies. As the spatial dependence is particularly difficult to characterize, the problem becomes how to use simple (i.e., biased) models to approximate the spatially dependent process. In particular, we propose the independent model and the Markov model as simple approximations. We conduct both theoretical analysis and extensive simulations on large networks using both real measurements and synthesized topologies to test the performance of the proposed models. Our results show that the independent model can capture temporal dependence and detailed topology information and, thus, outperforms the previous models, whereas the Markov model incorporates a certain spatial dependence and, thus, achieves a greater accuracy in characterizing both transient and equilibrium behaviors of malware propagation. PMID- 16252835 TI - Clinico-immunologic evaluation of allergy to Himalayan tree pollen in atopic subjects in India--a new record. AB - Exposure to local pollen allergens has a direct bearing on the prevalence of allergic symptoms among the inhabiting atopic population. The populations in the Himalayas and around it are exposed to a variety of pollen grains from trees growing in the region, but the pollen-population interaction has not been clinically investigated. Himalayan tree pollen from five different taxa, i.e. Alnus nitida (AN), Betula utilis (BU), Cedrus deodara (CD), Mallotus phillipensis (MP) and Quercus incana (QI) were evaluated for their allergenicity in the Indian population by in vivo (skin prick test) and in vitro (ELISA) clinico immunological methods. The presence of specific IgE against these tree pollen in the sera of skin test positive patients was taken as evidence for sensitization to these pollen. The average skin positivity in atopic populations recorded at different allergy centers in India varied from 2.2% against AN, to 4.7% against MP pollen. Significantly raised specific IgE against these pollen were observed in the sera of hypersensitive patients. The sensitization pattern to Himalayan tree pollen in these atopic populations varied. It was concluded that skin prick test positivity and raised IgE antibodies specific to AN, BU, CD, MP and QI established Himalayan tree pollen as important sensitizers in the atopic populations of India. A high incidence of skin sensitivity was observed to pollen antigens of Cedrus deodara, Mallotus phillipensis and Quercus incana in patients of Chandigarh residing in the hills and foothills of the Himalayas while Alnus nitida, Betula utilis and Cedrus deodara were important sensitizers in Delhi patients. The skin sensitization pattern against these pollen was in accordance with the level of exposure to these pollen of the subjects residing in that part of the country. PMID- 16252837 TI - Assessment of indoor air fungi in Western-Anatolia, Turkey. AB - This study was conducted to determine fungal spores in the indoor air of the houses in the city of Afyon, Western-Anatolia, Turkey. We investigated the seasonal properties of mould spores in 10 houses of Afyon over a period of one year. Viable moulds were recovered from all 10 houses. Twenty seven different moulds were isolated and identified from the indoor air of the houses. The most common genus was Cladosporium spp. (31.9%), followed by Aspergillus spp. (18.6%), Penicillium spp. (15.5%), Altemaria spp. (13.0%) and other species (21.0%). The mould concentration was higher in the kitchens than in other parts of the houses such as the living rooms and bedrooms (p < 0.05). The fungal flora of the air in the Afyon city region has a seasonal variation. All fungal species had their highest prevalence in summer and their lowest in winter, but only Aspergillus spp. had a significant seasonal variation (p = 0.012). Viable moulds are common in the houses of Afyon. Reducing these indoor fungi is necessary to improve the health of individuals with fungal-induced diseases like asthma. PMID- 16252836 TI - Detection of fungi spectrum in industrial and home bakeries and determinated fungal allergy with skin prick test. AB - Airborne fungal pathogens such as Penicillium, Aspergillus, Cladosporium, Trichophyton, and Alternaria may cause health problems. In this research, the fungal flora at different bakeries and their potential allergenic effects on the workers were investigated. We investigated 148 workers at 17 industrial type bakeries and 62 workers at 17 home type bakeries in Afyon. Our study was performed in two different seasons and climates, between January 2004 and June 2004. Fungal flora was detected by using Petri-dish method. In the winter, Penicillium was the dominant genus, while Cladosporium was the dominant genus during the summer, in both types of bakeries. The allergenic properties of dominant culturable fungi on workers involved in the bakeries were determined with the skin-prick test. It was found that with workers in the industrial type bakeries, the most common skin test positivity was caused by Penicillium. In the other hand, the skin test positivity, performed on workers in the home type bakeries, was equally caused by Penicillium, Trichophyton and Aspergillus. PMID- 16252839 TI - Malassezia furfur in infantile seborrheic dermatitis. AB - Our objective was to study both incidence and various strains of Malassezia in infantile seborrheic dermatitis (ISD). Sixty infants between 2 weeks and 2 years old with clinical diagnosis of ISD at the Department of Pediatrics, King Chulalongkorn Memorial Hospital from May 2002 to April 2003 were recruited. Malassezia spp. were isolated from cultured skin samples of the patients, genomic DNA was extracted and the ITS1 rDNA region was amplified. The PCR product was examined by agarose gel electrophoresis and DNA sequences were determined. The ITS1 sequences were also subjected to phylogenetic analysis and species identification. ISD is most commonly found in infants below the age of 2 months (64%), followed by those between 2 and 4 months (28%) old. Cultures yielded yeast like colonies in 15 specimens. PCR yielded 200-bp products (Candida) in 3 patients and 300-bp products (Malassezia furfur) in 12 patients (18%). Sugar fermentation using API 20C aux performed on the three 200-bp PCR products yielded Candida species. M. furfur was the only Malassezia recovered from skin scrapings of children with ISD. PMID- 16252838 TI - Quantitative analysis of antigen specific IgE in tears in comparison to serum samples. AB - We determined pollen specific IgE in tears and compared these results to the concentration of specific IgE in serum samples. We obtained tears (using Schirmer strips) and serum samples from subjects with Japanese cedar (Cryptomeria japonica) pollinosis, and tested for C. japonica pollen specific IgE using a quantitative ELISA. Time kinetic analyses through the pollen season showed that specific IgE levels in tears were found to increase earlier than those in sera and reached their maximum at the end of or after the pollen season, from March to early June. In the C. japonica pollen free season, July to December, the specific IgE levels in tears decreased, although the serum levels remained relatively high. These results indicate that the quantitative assay for specific IgE in tears might be useful to identify specific eye allergens. PMID- 16252840 TI - Increased smooth muscle actin expression from bone marrow stromal cells under retinoic acid treatment: an attempt for autologous blood vessel tissue engineering. AB - Vascular replacement in vital organs is sometimes necessary for human life for example because of atherosclerosis. Blood vessel tissue engineering is applied for autologous transplantations to avoid graft rejections. Stem cells are used for blood vessel tissue engineering because they are the origin of smooth muscle cells, endothelial cells and fibroblasts. This paper shows that bone marrow stromal cells (BMSCs) can be induced to differentiate into the early stage of smooth muscle cells by using 0.01 microM retinoic acid. The differentiation of BMSCs to smooth muscle cells was detected by the expression of smooth muscle alpha actin (SM alpha-actin), the earliest smooth muscle cell marker. The SM alpha-actin marker expression was demonstrated using indirect immunofluorescence technique and Western blot analysis. The induction of BMSC to form early stages of smooth muscle cells in this study is appropriate for blood vessel tissue engineering because the early stage smooth muscle cells may be stimulated to develop vascular walls with endothelial cells using a co-culture system. PMID- 16252841 TI - Serum nitric oxide in children with dengue infection. AB - One hundred and ten patients (M/F = 67/43) from King Chulalongkorn Memorial Hospital and the provincial hospitals of Uttaradit, Ayudhaya, and Sakonnakorn, who were clinically diagnosed with dengue infection and serologically confirmed by ELISA anti-Dengue IgM and IgG were recruited. Their serum NO level was measured using commercially available assay kits to investigate its correlation with the severity of the dengue infection: dengue fever (DF), DHF I/II, and DHF III/IV or dengue shock syndrome (DSS). Serum NO levels were also measured in 38 healthy controls (M/F = 19/19). Serum NO levels in dengue patients were lower than those of the controls (control = 168.18 +/- 24.10 micromol/l, DF = 124.94 +/ 36.79 micromol/l, DHF I/II = 99.69 +/- 33.42 micromol/l, and DHF III/IV = 120.63 +/- 46.26 micromol/l; p < 0.05). Serum NO levels in patients with DHF I/II were significantly lower than in those with DHF III/IV. These preliminary data revealed that levels of serum NO in dengue patients were significantly lower than those of normal controls. Patients with DSS had higher NO levels than those with DHF I/II. The decreased NO in dengue patients could be due to endothelial damage rendering the endothelium incapable of producing NO. Endothelial function seems to play a role in the pathogenesis of dengue infection. Further studies are required to see whether serum NO levels could play a role in the course of the disease and could help predict the severity of dengue infection. PMID- 16252842 TI - Restrictive lung disease and serum TGF-beta1 in thalassemia major children. AB - A cross sectional study was performed in 21 thalassemia major (TM) children at King Chulalongkorn Memorial Hospital during March to August, 2003 to determine whether restrictive lung disease (RLD) was related to serum transforming growth factor-beta 1 (TGF-beta1). All studied patients (57% female, age 11.2 +/- 2.6 yrs, duration of transfusion 7.7 +/- 4.1 yrs) never had desferoxamine treatment and their pulmonary function, serum ferritin and serum TGF-beta1 were evaluated. Five (24%) had RLD. RLD patients had significantly longer durations of transfusion and higher serum ferritin levels than non-RLD patients (9.1 +/- 1.9 vs 5.5 +/- 3.2 yrs; p = 0.03 and 3,816.6 +/- 1,715.9 vs 2,084.5 +/- 1,504.8 ng/ml; p = 0.04, respectively). TM children had lower serum TGF-beta1 levels than normal children (7.9 vs 78.8 pg/ml; p < 0.001). The serum TGF-beta1 level was not different between RLD and non-RLD patients (13.3 vs 4.2 pg/ml; ns), concluding that RLD was related to longer duration of transfusion and higher serum ferritin but not related to serum TGF-beta1 levels. PMID- 16252843 TI - Serodiagnosis of melioidosis by a competitive enzyme-linked immunosorbent assay using a lipopolysaccharide-specific monoclonal antibody. AB - Burkholderia pseudomallei is the causative agent of melioidosis, a severe and potentially fatal infectious disease in humans known to be endemic in Southeast Asia and northern Australia. The infection is also increasingly recognized in various animal species with a potential to spread to humans. With the potential as a biological warfare agent, specific serodiagnosis of melioidosis for surveillance in large populations at risk, humans or animals, would be highly valuable. In this study, a competitive enzyme-linked immunosorbent assay (ELISA) using a lipopolysaccharide-specific monoclonal antibody was developed. The assay provides high specificity, based on a previously described monoclonal antibody to a specific epitope on the lipopolysaccharide (LPS) of B. pseudomallei. The assay sensitivity of 96.0% and specificity of 100% were achieved at a cutoff value of 50% inhibition in human culture-proven melioidosis cases. An optimal cutoff value of 65% inhibition for sera from a melioidosis endemic area was obtained by ROC analysis and resulted in an assay specificity of 86.2%, while maintaining assay sensitivity of 92.0%. A potential application of the assay in the serodiagnosis of melioidosis in animal species was also evaluated usina dolphin sera with satisfactory results. PMID- 16252844 TI - LipL32, an outer membrane protein of Leptospira, as an antigen in a dipstick assay for diagnosis of leptospirosis. AB - Microscopic agglutination test (MAT), as well as other serological assays that aimed at detecting antibodies to Leptospira, supplements the leptospirosis diagnosis based on the clinical features. Nevertheless, false positive results have been occasionally reported when the crude antigen was used in those antibody based tests due either to the presence of antibodies stimulated by other antigenically related pathogens in the patient's serum, or the antibodies in the serum may be stimulated by a previously unrecognized Leptospira infection, especially in the disease endemic areas. Thus, the more refined antigen should improve the serodiagnostic accuracy. Among Leptospira spp., LipL32, which is a pathogenic Leptospira outer membrane protein (OMP), expressed by the bacteria grown both in vitro and in vivo. In this study, recombinant LipL32 protein was tested by a dipstick method for its potential in serodiagnosis of leptospirosis. Preliminary results suggest that the recombinant LipL32 is a good diagnostic detection reagent for specific Leptospira IgG. Diagnostic sensitivity and specificity of the Lip32 dipstick assay, when compared to those of MAT, were 100% and 98.33%, respectively. PMID- 16252845 TI - HIV-1 subtyping using gag/env heteroduplex mobility assay and peptide enzyme linked immunosorbent assay. AB - Two HIV-1 subtypes have accounted for virtually all infections in Thailand: subtype B', found mainly in injection drug users (IDUs), and CRF01_AE (initially subtype E), found in over 90% of sexually infected persons and increasingly in IDUs in recent years. During 1997-1998, 227 blood samples were collected from HIV 1 infected individuals consisting of 92 mothers, 35 children and 100 IDUs. The blood samples were subtyped by heteroduplex mobility assay (HMA) and peptide enzyme-linked immunosorbent assay (PEIA). Using gag and env HMA, CRF01_AE and subtype B' accounted for 96-97% and 3-4% of both the mothers and the children, respectively. In the IDU group, 10% of the plasma samples could only be performed by gag HMA and gave the result as CRF01_AE. CRF01_AE and subtype B' using PEIA accounted for 67% and 33% of the IDUs. There was 100% concordance of the results between gag HMA and env HMA. Ninety-five percentages of concordant results were observed between HMA and PEIA. Of the 6/134 (5%) subjects with discordant results, nucleotide sequencing, used as a gold standard, confirmed the HMA result. In this study, HIV-1 was successfully genotyped by HMA and PEIA. However, a comparison of the subtyping results between HMA and PEIA revealed that HMA was slightly more accurate than PEIA. PMID- 16252846 TI - Prevalence of positive antinuclear antibodies in healthy children. AB - Antinuclear antibodies (ANA) frequently arise in the sera of children with connective tissue disease and is used in the diagnosis of these diseases. Therefore it is also important to know the prevalence of ANA in normal children. The main objective of the present study was to determine the prevalence of antinuclear antibody (ANA) in healthy children. Ninety-nine serum samples from a serum bank and 108 samples from patients who had attended elective surgery and whose blood had been withdrawn for other investigations, were tested for ANA by indirect immunofluorescence method using HEp-2 cells as substrate. Sera from 52 children with SLE were also tested during the same period. It was found that antinuclear antibodies were present in 32 (15%) of the 207 sera of healthy children at a dilution of 1:40 or higher. ANA were positive in 9% at a serum dilution of 1:40, in 3% at 1:80 and in 3% at 1:160. The patterns of immunofluorescence staining were as follows: homogeneous in 46.7%, speckled in 20%, and nucleolar in 10%. In SLE patients, ANA were positive in 91%; 13% at a serum dilution of 1:40, 7% at 1:80, 20% at 1:160, 15% at 1:320, 9% at 1:640, 20% at 1:1,280 and 9% at > or = 1:2,560. It was concluded that the prevalence of positive ANA using the HEp-2 cells as substrate was 15% in healthy children at dilutions of 1:40 or higher. Using the cutoff serum dilution of 1:40, the sensitivity of this test was 91%, the specificity was 85%, the positive predictive value was 57% and the negative predictive value was 97%. PMID- 16252847 TI - DiGeorge syndrome associated with solitary median maxillary central incisor. AB - DiGeorge syndrome is a primary immunodeficiency disease characterized by dysgenesis of the thymus and parathyroid glands, conotruncal cardiac anomalies, and other dysmorphic features. Although most patients have a common microscopic deletion in chromosome 22q11.2, marked clinical variability exists. A solitary median maxillary central incisor (SMMCI) is a rare dental anomaly which may be an isolated occurrence or associated with congenital nasal airway abnormalities or holoprosencephaly. We report a patient with DiGeorge syndrome who was diagnosed at nearly 1 month of age and was later found to have a solitary median central incisor. Initially, the patient presented with recurrent episodes of respiratory distress attributed to partial airway obstruction, one of the phenotypic features of SMMCI. A fluorescence in situ hybridization study showed a chromosome 22q11.2 deletion. PMID- 16252848 TI - Raising rheumatoid factor cutoff helps distinguish rheumatoid arthritis. AB - The presence of rheumatoid factor (RF) is one of the clinical criteria for the diagnosis of rheumatoid arthritis (RA). The cutoff point of RF assays is usually based on a reference level obtained from normal subjects in the same population as the patients. We evaluated 63 rheumatoid arthritis (RA), 25 other arthritis patients and 110 blood donors. Their rheumatoid factors (RF) ranged from < 9.9 to 2,264, < 9.9 to 262, and < 9.9 to 66 mIU/ml, respectively. The sensitivity at different cutoff points of 15, 20, and 25 mIU/ml was 92.1%, 90.5%, and 88.9%, respectively. The specificity at the same cutoff points was 81.5%, 84.4%, and 85.2%, respectively. Having minimally sacrificed the sensitivity, we recommend using a higher RF cutoff to increase specificity. PMID- 16252850 TI - [Population structure and community characteristics of Pseudotaxus chienii in Fengyangshan National Natural Reserve]. AB - Pseudotaxus chienii, an endemic plant in China, is one of the second grade state protection wild plants, and distributes in Fengyangshan Natural Reserve as one of its concentrative dwelling places. A survey in the region was carried out on 10 communities, which were dominated by P. chienii. The analysis on its size structure, spatial distribution pattern and community characteristics showed that the populations of P. chienii could grow in the communities dominated by Rhododendron simiarum, Fokienia hodginsii-R. simiarum, and evergreen broad-leaved forests. In R. simiarum communities, the size structure, survival curve, and overwhelming community distribution pattern of P. chienii showed a sustaining development, while in communities dominated by F. hodginsii-R. simiarum, though the size structure was declining, the survival curve was Deevy- III type. Plenty of plantlets were still existed, and the populations kept steady. Two types were considered to be the most suitable ones for P. chienii populations. The important value of P. chienii reached 5% - 10%. In evergreen broad-leaved forests, the populations of P. chienii showed to be a declining type, and the survival curve was Deevy- II type. The distribution pattern was random, indicating that P. chienii populations could not fit for survive in this kind of communities. Correlation analysis showed that P. chienii populations had a positive correlation with R. simiarum and F. hodginsii, but a negative correlation with Cyclobalanopsis stewardiana and Schima superba. P. chienii could associate to the habits with the community canopy density of about 0.6 - 0.8. From the results mentioned above, the populations of P. chienii could be able to sustain and develop, and the existing habitat in Fengyangshan should be protected effectively. The protection of P. chienii requires more basic work to establish efficient measures to protect its habitat. PMID- 16252849 TI - [Tree species diversity of tropical montane rain forest in Xishuangbanna, Yunnan]. AB - Tropical montane rain forest (TMR), one of the main forest vegetation types in Xishuangbanna of Yunnan Province, occurs horizontally in the north (TMRF I) and vertically in the south (TMRF II). In order to understand its tree species diversity in different zones, six fixed plots (50 m x 50 m) at different sites in Xishuangbanna were established, two (plots 1 and 2) for TMRF I, and four (plots 3, 4, 5 and 6) for TMRF II. Grid method (10 m x 10 m) was used to record all the individuals with DBH greater than 2.0 cm in each plot, and tree species richness and diversity characteristics along DBH class, as well as changing trend of tree species diversity with increasing sampling area were analyzed. The results showed that the tree stem and species richness in the six plots both declined constantly along the DBH class gradient. For the communities of TMRF I (altitude was 850 1000 m), all indices had no clear trend with the gradient, while for the communities of TMRF II (altitude 1 200 - 2000 m), Shannon-Wiener and Simpson indices declined constantly with gradient, but Pielou's evenness index had the opposite trend. In smaller DBH classes (> or =2 cm, > or =5 cm and > or =10 cm), the tree species richness, diversity, and evenness of the communities of TMRF II were obviously higher than those of the communities of TMRF I, but there was no difference between the communities of TMRF I and TMRF II in larger DBH classes (> or = 20.0 cm, > or =30.0 cm and > or =50.0 cm) . With sampling area increased, the tree species richness, Shannon-Wiener index, Simpson index and Pielou's evenness index of the six plots all had the trend to be smooth when the sampling area increased to 2 000 m2. PMID- 16252851 TI - [Root biomass and underground C and N storage of primitive Korean pine and broad leaved climax forest in Changbai Mountains at its different succession stages]. AB - With more than 200 years old primitive Korean pine and broad-leaved climax forest and its two 20 and 80 years old secondary Populus davidiana and Betula platyphylla forests in Changbai Mountains as test objects, this paper studied their root biomass and underground C and N storage. The results showed that with forest succession, the root biomass of 20 years old, 80 years old, and climax forests was 2.437, 2.742 and 4.114 kg x m(-2), respectively. The root C storage was 1.113, 1.323 and 2.023 kg x m(-2), soil C storage was 11.911, 11.943 and 12.587 kg x m(-2), and underground C storage was 13.024, 13.266 and 14.610 kg x m(-2), respectively, while the root N storage was 0.035, 0.032 and 0.038 kg x m( 2), soil N storage was 1.207, 1.222 and 0.915 kg x m(-2), and underground N storage was 1.243, 1.254 and 0.955 kg x m(-2), respectively, which indicated that along with forest succession, forest underground became a potential "carbon sink", whereas underground N storage did not change obviously. PMID- 16252852 TI - [Temporal and spatial dynamics of reproductive modules of endangered plant Disanthus cercidifolius var. longipes natural populations]. AB - Based on field observation data, this paper studied the flowering phenology and the temporal and spatial dynamics of the reproductive modules of endangered plant Disanthus cercidifolius var. longipes natural populations. The results showed that there were certain differences of the flowering phenology among the populations in different communities, e. g., in pure forest, the average flowering duration of individuals was the longest (75 d), while the flowering synchrony was the lowest (0.717 d), but in bamboo forest, the flowering duration of individuals was the shortest (47 d), while its flowering synchrony was the highest. The numbers of reproductive branches and inflorescences were increased with increasing individual age, and reached the highest at the age of 30 - 35 yr. At the elevation of 810 m, the numbers of individual reproductive branches and inflorescences reached the peak, and there were significant differences among the crown levels. The reproductive branch number of individuals and the inflorescences per reproductive branch were the highest in pure forest, being 411.39 and 7. 857, respectively, while the inflorescences per reproductive branch in bamboo forest were notably lower than those in other communities. The abortive ratio of reproductive modules of young individuals was higher than that of the older ones in the flower-bud, flowering, and fruit phases. The abortion of reproductive modules had no correlation with their distributed elevation, but some correlations existed with their development time, distributed crown levels and communities. The major factor affecting the abortive ratio of individual reproduction modules was probably the illumination condition in the distributed communities. PMID- 16252853 TI - [Characteristics of Robinia pseudoacacia water physiological ecology under different habitats in North Shaanxi gully areas of Loess Plateau]. AB - With locust (Robinia pseudoacacia), the main tree species in afforestation, as test material, this paper studied the characteristics of its water physiological ecology and productivity under four habitats, i. e. , sunny, shady, semi-sunny and semi-shady hillsides, in the North Shaanxi gully areas of Loess Plateau. The mean water content in 0 - 500 cm soil layer was 8.87% (shady), 8.06% (semi shady), 7.62% (semi-sunny), and 6.96% (sunny), respectively. There was a significant discrepancy between shady, semi-shady, semi-sunny and sunnyhilisides (a 0.01), and the difference between shady, semi-shady and semi-sunny, as well as between semi-sunny and sunny hillsides was also significant (a 0.05) . Remarkable relationships were found between leaf RWC, locus WSD and soil water content (SWC) . The leaf RWC and locust water potential under sunny hillside were lower, but those under shady hillside were higher. The daily mean transpiration was in order of shady (4.07 microg x cm(-2) x s(-1)) > semi-shady (3.89 microg x cm(-2) x s( 1)) > semi-sunny (3.05 microg x cm(-2) x s(-1)) > sunny (2.70 microg x cm(-2)s( 1)) hillside. The remarkable difference of transpiration appeared at 11:00 and 13:00, and there existed a remarkable relationship between transpiration and light intensity, RH and soil water content. All of these resulted in a diversity of locust biomass under different habitats, the highest under shady hillside, and the lowest under sunny hillside, and the differences between different habitats were all significant. It could be concluded that soil water content was the main factor affecting locust growth. PMID- 16252854 TI - [Relationship between species distribution and habitat effective temperature in subalpine meadow of Mt. Xiaowutai]. AB - By the methods of sugar-inversion, plant community field survey and laboratory analysis, this paper measured the effective temperature of habitats, relative coverage of species, and aboveground biomass in subalpine meadow of Mt.Xiaowutai, and analyzed the relationship between species distribution and habitat effective temperature in landscape scale. The results showed that among 97 vascular plant species registered, the distribution patterns of about 36 % of total species were influenced by the effective temperature of the habitats, and the relative coverage of about 20% of them correlated significantly with the effective temperature. Eight species showed a negative correlation between their relative coverage and effective temperature, and eleven species had a positive correlation between their relative coverage or aboveground biomass and effective temperature. The eleven species with a positive correlation had certain drought resistance feature or a tendency distributing towards warmer habitats. Concerning the effectiveness of species feature applied in the analysis, relative coverage could be used to determine the relationship between species distribution and effective temperature efficiently, if the species frequency ranged from 90 % to 20 %. But, for the dominant species with their frequency over 90 %, aboveground biomass seemed to be more suitable for the analysis. PMID- 16252855 TI - [Estimation models for vegetation water content at both leaf and canopy levels]. AB - Based on spectral indices method, this paper utilized the water content (Cw) and reflectance data of 67 fresh different type leaves from LOPEX' 93 database to establish the statistical model between leaf Cw and spectral indices at leaf level through 47 samples, and to test the model with the other 20 samples. The results suggested that fuel moisture content (FMC) and equivalent water thickness (EWT) as Cw demonstrators were different in reflectance spectral curves. The difference between FMC and EWT was large when they were utilized to retrieve the leaf Cw. The correlation coefficient between EWT and each spectral index was higher than FMC, but the forecast precision of FMC was better than that of EWT. The 7 spectral indices could all retrieve the leaf FMC accurately, but only the Ratio975, II and SR were suitable to estimate the leaf EWT. Spectral indices linear model on the strength of optimal subset regressions had the highest precision to retrieve the leaf Cw. Ratio975 might be the universal spectral index to estimate the leaf Cw. At canopy level, the simulated canopy spectra under different leaf area index (LAI) and Cw were derived from the PROSPECT and SAILH coupling models. In order to eliminate background influence and to precisely retrieve the Cw, soil-adjusted water index (SAWI) was proposed at the first time to indicate the information of near-infrared and short-wave infrared canopy reflectance. The ratio of SAWI and other spectral indices could dramatically eliminate the soil background, and effectively retrieve the vegetation Cw at canopy level. Spectral index (Ratio975 - 0.96)/(SAWI + 0.2) as improved Ratio975 could be used to compute the canopy Cw more precisely when LAI was ranging from 0.3 to 8.0 and Cw from 0.0001 to 0. 07cm. PMID- 16252856 TI - [Haloxylon ammodendron community patterns in different habitats along southeastern edge of Zhunger basin]. AB - Low-lying land, slow and gentle desert, and semi-mobile dune are the three different habitats of natural Haloxylon ammodendron community along the southeastern edge of Zhunger Basin in Xinjiang Uygur Autonomous Region. This paper studied the structural characters of H. ammodendron community from the aspects of species structure, species diversity, biomass, and distribution patterns of natural regeneration sapling. The results showed that the species of H. ammodendron community was the richest on low-lying land, the second on slow and gentle desert, and the least on semi-mobile dune. The number of plant species in the three different habitats was 16, 15 and 12, respectively. The amount of H. ammodendron natural regeneration sapling was the largest (6 687 trees x hm(-2)) on semi-mobile dune, but its distribution was not even. Low-lying land had a slightly smaller amount (5 799 trees x hm(-2)) of H. ammodendron natural regeneration sapling than semi-mobile dune, but the distribution of the sapling was more even. The overall evaluation ion was that the natural regeneration of H. ammodendron community was the best on low-lying land. Its total biomass on low lying land was 19.39 t x hm(-2), while that on slow and gentle desert and semi mobile dune was 9.32 and 6.69 t x hm(-2), respectively. The distribution patterns of H. ammodendron natural regeneration sapling in different habitats were all aggregatice. The ground of low-lying land was fixed, with fairly good soil moisture and fertility, which was appropriate for the growth of H. ammodendron and the development of H. anmmodendron community, while that of slow and gentle desert and semi-mobile dune was easier to suffer from wind erosion, with poor soil moisture and fertility and fairly serious habitat conditions. PMID- 16252857 TI - [Phosphorus and sulphur bio-cycling in alpine tundra ecosystem of Changbai Mountains]. AB - The study with compartment model on the phosphorus (P) and sulphur (S) bio cycling in alpine tundra ecosystem of Changbai Mountains showed that the total storage of P and S was 16 088.6 t and 26 079.4 t, of which, 46.14 t and 64.82 t was in vegetation pool, 89.63 t and 53.16 t in litterfall pool, and 15952.8 t and 26014.6 t in soil pool, respectively. The above- and below-ground vegetation pool stored 21.88 t and 44.21 t, and 24.28 t and 20.61 t of P and S, respectively, and the above-ground vegetation pool had 47.4% of P and 68.2% of S in the vegetation subsystem. The transferable P amount was 24.25 t x yr(-1) through plant absorption and 31.59 t x yr(-1) through litterfall return, while the transferable S amount was 31.18 t x yr(-1), 10.12 t x yr(-1) and 21.06 t x yr(-1) in the aboveground plant, belowground root system, and litterfall return, respectively. The natural return ratio of S was 67.5%. PMID- 16252858 TI - [Early responses of soil fauna in three typical forests of south subtropical China to simulated N deposition addition]. AB - In this paper, simulated N deposition addition (0, 50, 100 and 150 kg x hm(-2) x yr(-1)) by spreading water or NH4NO3 was conducted to study the early responses of soil fauna in three typical native forests (monsoon evergreen broadleaf forest, pine forest, and broadleaf-pine mixed forest) of subtropical China. The results showed that in monsoon evergreen broadleaf forest, N deposition addition had an obviously negative effect on the three indexes for soil fauna, but in pine forest, the positive effect was significant (P < 0. 05), and the soil fauna community could reach the level in mixed forest, even that in monsoon evergreen broadleaf forest at sometime. The responses in mixed forest were not obvious. In monsoon evergreen broadleaf forest, the negative effects were significant (P < 0.05) under medium N deposition, but not under low N deposition. In pine forest, the positive effect was significant (P < 0.05) under high N deposition, especially for the number of soil fauna groups. The results obtained might imply the N saturation-response mechanisms of forest ecosystems in subtropical China, and the conclusions from this study were also consisted with some related researches. PMID- 16252859 TI - [Ecological characteristics and modulate strategy of wintertime climate in self controlled greenhouse]. AB - Based on the five years observation data in self-controlled greenhouse at Donghai vegetable demonstration base in Shanghai, and adopting mathematical statistics methods, this paper analyzed the monthly variation characteristics of ambient illumination, temperature and CO2 concentration during wintertime, disclosed the persisting hours of favorable and unfavorable illumination grade under different weather conditions, appearing cases of different threshold air temperature, persisting hours and characteristics of CO2 shortfall, and variation characteristics of air relative humidity. The correlation equations between daytime average air temperature, CO2 concentration, relative humidity and illumination in greenhouse were set up, the main problems in self-controlled greenhouse during wintertime were pointed out, and some related modulate advises were put forward. In Shanghai area, the key points of obtaining high yields in greenhouse during wintertime should be to insure sufficient illumination, modulate air temperature at nights, raise forenoon air temperature in sunny days, and enlarge air temperature difference between daytime and night to increase net photosynthesis matters. PMID- 16252860 TI - [Comprehensive management patterns of economic fruit forest in Dashan Village and their ecological and economic benefits]. AB - With the theories of economy and ecology, this paper analyzed the main management patterns of economic fruit forest in the Dashan Village, Huoqiu County, Anhui Province. The results showed that 9-year-old mandarin-tea, 5-year-old persimmon potato-watermelon, 7-year-old pear-potato-soybean, and 7-year-old pear balloonflower had obvious social, economic and ecological benefits. Their net economic benefits were 8 700.00, 12 351.00, 12 337.50 and 22 500.00 yuan x hm( 2), respectively, higher than that of single crop planting. In these patterns, the crown density between rows could reach 0.3 - 0.4, and the light utilization rate increased by 20% - 30%. PMID- 16252861 TI - [Effects of rice-duck farming system on Oryza sativa growth and its yield]. AB - The study showed that under rice-duck farming, the number of rice non-productive tiller reduced significantly, the ratio of its effective panicles increased by 8.08 %, and its basal penetration light rate enhanced by 4.05%. At full-heading and maturing stages, the green leaf ratio under rice-duck farming was 6.01% and 10.65% higher than the control, and the leaf chlorophyll content was increased by 2.90% and 17.82%, respectively. Under rice-duck farming, the root vigour at full heading stage and the photosynthesis capability of flag leaf at grain-filling stage were increased by 24.2% and 15.73%, which could accumulate more assimilative matter, and increase the harvest index and yield by 2.87% and 4.93%, respectively. PMID- 16252862 TI - [Effects of genotype and environment on protein and starch quality of wheat grain]. AB - A field trial with 10 winter wheat varieties was conducted at 9 locations under different environmental conditions in Shandong Province to study the effects of wheat genotype and environment on the main qualitative characters of wheat grain. The results indicated that variety and environment had a marked influence on the main qualitative indexes of wheat grain, and the difference of environmental conditions was the main factor affecting the protein and gluten contents of the grain. The variation of farinogramme indexes was mainly due to the interaction between variety and environment. As for the starch content, peak viscosity, and breakdown of the grain, interactive effect was stronger than independent one, and the effect of variety was stronger than that of environment. PMID- 16252863 TI - [Effects of osmotic stress on chlorophyll fluorescence parameters of wheat seedling]. AB - By using chlorophyll fluorescence kinetics technology, this paper determined the chlorophyll fluorescence parameters Fv/Fm, phiPSII, qP, qNP, ETR of winter wheat seedling under simulated osmotic stress. The results showed that with the increase of osmotic stress, Fv/Fm and Fv/Fo had the trend of decrease-increase decrease, indicating that photoinhibition did not happen at the first stage of osmotic stress, but the followed increase of Fv/Fm resulted in the photoinhibition and the decrease of phiPSII and ETR. During the course of osmotic stress, qP and qNP decreased firstly and then increased, which was helpful to enhance the open ratio of PSII reaction center, and made more photosynthetic energy use the photosynthetic electron transport to enhance the electron transport capacity. Meanwhile, the increase of non-photochemical quenching coefficient could be helpful to dissipate excess light energy, which protected the photosynthetic tissue, and mitigated the effect of environment on photosynthesis. All the results indicated that wheat seedling had its protection mechanism. Between the two test winter wheat varieties, Changwu 134 had a higher drought-resistance capacity than Shan 253. PMID- 16252864 TI - [Photo-thermal characteristics of a non-photosensitive and extra-premature winter wheat variety]. AB - In a sowing by stages test with winter wheat variety Jingdong 8 (JD8) as reference, this paper studied the photothermal characteristics of a non photosensitive and extra- premature winter wheat variety Dongzao 5 (DZ5), and the effects of sowing stages on its growth and yield. The results showed that the harvest date of DZ5 was 4-5 days earlier than that of JD8, and its yield with standard sowing date increased by 43.4%. In addition, DZ5 had a shorter thermoperiod for ear differentiation, and didn't need strict vernalization process and photoperiod, which could be sown either before or after winter. PMID- 16252865 TI - [Dynamics of soil active organic matter in Chinese fir plantations]. AB - Soil active organic matter is the main source of soil nutrients, and plays an important role in the formation and stabilization of soil aggregate. Chinese fir (Cunninghamia lanceolata) is the most important fast-growing timber tree species in southern China, but its continuous plantation has caused soil deterioration. The study on the active fractions of soil organic matter under the first and second generation plantations of Chinese fir and the native broad-leaved forest at Huitong Experimental Station of Forest Ecology, Chinese Academy of Sciences, in Hunan Province showed that their contents were higher under native broad leaved forest than under Chinese fir plantations, and higher under the first than under the second generation plantation. The content of soil active organic carbon (SAOC), microbial biomass carbon (MBC), water-soluble organic carbon (WSOC) and water-soluble carbohydrate (WSC) was 18.79 g x kg(-1), 421.7 mg x kg(-1), 252.2 mg x kg(-1) and 136.3 mg x kg(-1) under the first generation plantation of Chinese fir, and 22.31 g x kg(-1), 800.5 mg x kg(-1), 361.1 mg x kg(-1) and 220.1 mg x kg(-1) under the native broad-leaved forest, respectively. The corresponding values under the second generation plantation of Chinese fir were 73.6%, 87.9%, 66.3% and 53.2% of those under the first generation plantation of Chinese fir. The results also indicated that there existed some extent correlations among the active fractions of soil organic matter, and higher correlations between soil microbial biomass carbon and other active fractions of soil organic matter. PMID- 16252866 TI - [Microbial diversity in continuously planted Chinese fir soil]. AB - The study on the microbial diversity of 1st, 2nd, 3rd and 4th generation Chinese fir soil at Huitong Forest Experimental Station showed that with the increase of generation, the numbers of bacteria and actinomyces decreased markedly, while that of fungi increased evidently. It was found by PCR and DGGE that the bacterial diversity changed slightly and its genetic similarity among different generations reached 87 %, but the fungal diversity reduced and its genetic similarity was only 45%. The numbers of pathogenic and deleterious fungi increased markedly with increasing Chinese fir generations. PMID- 16252867 TI - [Effects of different de-farming and reafforestation patterns on changes of soil fertility quality in Karst region of southwestern China]. AB - A ten-year fixed site harnessing was conducted in the severely eroded karst region of Ziyun County, Guizhou Province by adopting four different de-farming and reafforestation patterns, and the properties of soil chemistry, soil microbiology, and soil enzymology after harnessing were determined in 2003. The results showed that the total amount of soil microbes and the individuals of bacteria, fungi and actinomyces were increased obviously, the activities of soil hydrolytic and oxidoreductive enzymes and the soil respiration rate enhanced evidently, and the storage of soil nutrients as well as their supplying intensities promoted pronouncedly, demonstrating that the soil quality in all adopted patterns was improved in varying degrees. The values of soil integrated fertility index (IFI) had an increasing trend. Therefore, proper biological measures and essential supplementary engineering measures were effective in improving the soil fertility quality of severely degraded karst region. PMID- 16252868 TI - [Spatial variability of slope land soil solute transport parameters]. AB - In this paper, the transport parameters of chlorine in a slope land soil were determined in situ by fitting breaking-though curve, and the spatial variability of the parameters in a 41 m x 5 m runoff plot was analyzed by using traditional and geo-statistics. The results showed that the average pore-water velocity increased gradually from the top to the bottom of the slope, and the diffusive dispersive coefficient (D) had no remarkable variation within 20m away from the top but gradually increased with the distance beyond 20m. The dispersivity (alpha) had the same characteristics as D. The pore-water velocity (v) had a floating feature on the slope, while parameters D and alpha showed obvious spatial correlation characteristics, the range of the spatial correlation being 21m and 10m, respectively. PMID- 16252869 TI - [Correlation between inhibition activity of endophytic fungus from Euphorbia pekinensis and its host]. AB - To develop endophytic fungi resources from medicinal plants, this paper studied the correlation between endophytic fungus Fusariium sp. ES from Euphorbia pekinensis and its host through analyzing the inhibition activity of this pathogen. An endophytic strain S12 (Alternaria sp.) from Sapium sebiferum was used as contrast. The results showed that E5 had an inhibitory effect on the host, while S12 and E. pekinensis extract didn't have it. The inhibition activity of E5 could be stimulated by the extract of host leaf, and its inhibitory effect could only be found when it was cultured with enough oxygen. The inhibition activity was affected by pH, temperature, and ultra-violet. These characters of endophytic fungus E5 showed a good correlation with its host, and this correlation was mutualism. PMID- 16252870 TI - [Effects of elevated CO2 concentration on rhizosphere soil microbes under Pinus koraiensis and Pinus sylvestriformis seedlings]. AB - The study showed that under Pinus sylvestriformis seedlings, the amount of rhizosphere soil bacteria, but not fungi and actinomyces, increased significantly (P < or = 0. 001) with CO2 enrichment (700 and 500 micromol x mol(-1)), and the same (P < or = 0.001) was under Pinus koraiensis seedlings, with the exceptions of 700 micromol CO2 x mol(-1) in August and 500 micromol CO2 x mol(-1) in July, compared to the ambient chamber and the ambient in each month. No significant effect of elevated CO2 was found on the amount of rhizosphere soil fungi except in September ( P < or = 0.001), and the response of actinomyces to elevated CO2 was also insignificant. PMID- 16252871 TI - [N2O emission factor for agricultural soils]. AB - Based on the direct measurements of annual N2O emission from 207 field experiments reported in literatures between 1982 and 2003, this paper established a database of annual N2O emission from agricultural fields. Correlation analysis indicated that the N2O emission from agricultural soils was significantly correlated with climatic factors temperature and precipitation, while no significant correlations were observed between N2O emission and edaphic parameters pH, organic carbon and nitrogen. According to the definition of N2O-N emission factor and its quantification by the Intergovernmental Panel on Climate Change, the N2O-N emission factor was modified by annual mean temperature and annual precipitation, respectively. The results suggested that the modification with precipitation might significantly reduce the estimated error of N2O emission by about 16%, while that with temperature did not reduce the error, in comparison with the default emission factor by the Intergovernmental Panel on Climate Change. PMID- 16252872 TI - [Effects of organic manures on CO2 and CH4 fluxes of farmland]. AB - This paper studied the effects of chicken manure, swine waste and cattle manure on the CO2 and CH4 fluxes of a farmland planted with summer maize. The results showed that the CO2 flux had the same trend under different organic manure application, which was influenced by soil temperature and soil water content. The flux was significantly related with air temperature, soil surface temperature and soil temperature (P < 0.05). When soil temperature was not a limiting factor, soil water content was significantly related with soil CO2 flux. At most maize growth stages, soil was a sink of CH4. The variation of CH4 source-sink was influenced by environmental factors, but the correlation was not significant. The mean seasonal flux of CO2 was from 0.5124 to 0.8518 g x m(-2) x h(-1), and that of CH4 was from - 0.0068 to - 0.0484 mg x m(-2) x h(-1). Compared with CK2, maize planting and organic manure application enhanced CO2 emission. The application of organic manures inhibited the CH4 uptake by soil, and higher application rate had a higher inhibitory effect. PMID- 16252873 TI - [Geostatistics analysis on spatial patterns of Myzus persicae and Erigonidium graminicola in plum orchard]. AB - Investigations on the spatial construction and distribution of Myzus persicae and Erigonidium graminicola in a plum orchard were conducted from March 2003 to November 2003. The results indicated that the semivariogram of Myzus persicae could be described by spherical model, except on June 27 and November 22, which should be described by lined model, and that of Erigonidium graminicola could be described by spherical model, except on May 21, May 31, October 19 and November 22, which should be described by lined model. It could be concluded that the amount and spatial distribution of Erigonidium graminicola was closely related to those of Myzus persicae. PMID- 16252874 TI - [Effects of corn cultivar on Tetranychus truncatus ehara (Acarina: Tetranychidae) population parameters]. AB - The laboratory study at 28 degrees C showed that on four different corn cultivars, the durations of ovum, nymph and pre-oviposition, adult longevity, fecundity, egg production per day, net reproductive rate (R0), finite rate of increase (lambda), intrinsic rate of increase (rm), mean generation time (T), and the days needed to double population size (DDP) of Tetranychus truncatus were significantly different. The hatching rate on Zea mays cv. Zhongdan 2, Nongda 108, Chidan 202 and Badan 3 was 95.8%, 94.0%, 90.0% and 84.0%, and the survival rate on these cultivars was 90.5%, 86.0%, 84.0%, and 72.0%, respectively. All the survival curves were of Deevey I . The net reproductive rate (R0) was the highest on Zhongdan 2, and the lowest on Badan 3. The mite displayed a decreasing preference in order of Zhongdan 2 > Nongda 108 > Chidan 202 > Badan 3. PMID- 16252875 TI - [Controlling effects of plant extracts and pesticides on Myzus persicae and Lipaphis erysimi populations]. AB - This paper studied the controlling effects of Xanthium sibiricum and Syngonium podophyllum ethanol extracts, mobile oil emulsion and 0.3% azadirachtin EC on Myzus persicae and Lipaphis erysimi populations both in laboratory and in field. The results showed that in laboratory, X. sibiricum extract was the key deterrent to Myzus persicae, while sprayed it with S. podophyllum extract and with petroleum spray oil or 0.3 % azadirachtin EC, the deterrent effect could be improved. As for pterigota Lipaphis erysimi, the extracts of the two plants were the main deterrents, and their interference effect was non-linear. In field, S. podophyllum extract had a stronger deterrent effect when sprayed independently. L. erysimi population could be successfully controlled by the mixtures of plant extracts and insecticides, with an efficiency of 95.7%. The best treatment against M. persicae population was X. sibiricum extract, with an efficiency of 87%, while S. podophyllum extract mixed with 0.3% azadirachtin EC, and X. sibiricum extract mixed with 0.3% azadirachtin EC had an efficiency of >80%. PMID- 16252876 TI - [Roles of olfaction and vision in orientation behavior of adult Campylomma chinensis Schuh (Hemiptera: Miridae) toward Lantana plants (Verbanaceae)]. AB - Investigations were carried out on the orientation behavior of adult Campylomma chinensis Schuh (Hemiptera: Miridae) toward plant hosts Lantana camara, L. caniara cv. 'Flava', and L. montevidensis (Spreng. ) Briq. (Verbanaceae). Surveys on three Lantana plants showed that the distribution of C. chinensis in inflorescences was not related to floral color (partial correlation coefficient was 0.240, P = 0. 147, n = 40), but to the number of Thrips hawiiensis Morgan (partial correlation coefficient was 0.512, P < 0.0001, n = 40) and the flowers per inflorescence (partial correlation coefficient was 0.451, P < 0.0001, n = 40). In a colour preference experiment, no adult C. chinensis attracted to six different colour plates in longan field. The testing results of four-armed olfactometer indicated that adult C. chinensis had a significantly higher preference for L. camara. It was concluded that olfactory stimuli played an important role in searching for plant hosts of C. chinensis. Solid phase microextraction (SPME) and GC-MS were employed to analyze the volatiles compounds of Lantana, and seven same chemical constituents were detected from the volatiles of three Lantana plants inflorescence. PMID- 16252878 TI - [Population dynamics of Bactrocera dorsalis (Diptera: Tephritidae) in Xishuangbanna of Southern Yunnan]. AB - In the study, an all year round monitoring through lure-traps in 1997, 2000 and 2003 was conducted on the population dynamics of oriental fruit fly, Bactrocera dorsalis (Diptera: Tephritidae) in Xishuangbanna of Southern Yunnan, and the factors in relation with its population fluctuation, including temperature, rainfall, and host species were analyzed. The fruit fly occurred year-round in Xishuangbanna, with a low population level from November to February, and increased steadily from March to July or August. An increasing peak appeared in July or August, depending on the rainfall amount in that year. Afterwards, the fly population declined markedly until to October. Temperature, rainfall, and host fruits were supposed to be the major factors affecting the population fluctuation. The monthly average temperature in Xishuangbanna was in the ranges suitable for the development and reproduction of the fly, but the monthly average minimum temperature from December to February appeared to be lower than the suitable temperature, which was suggested to be responsible for the low populations in the winter months. Rainfall was another essential factor affecting population fluctuation. The population was depressed when the monthly rainfall amount was lower than 50 mm, but increased when the amount ranged between 100 and 200 mm. Under conditions the monthly rainfall amount was higher than 250 mm, the fruit fly population was reduced markedly, which explained the decreasing population in July or August when the heavy rain occurred. Mango, orange, pear, longan, and peach were the main host species of the fly, among which, mango and longan were most preferred by the fly. Therefore, their planted area, fruiting period and productivity exerted essential effects on the fly population fluctuation, and regarded as another major factor affecting the fly population in this area. PMID- 16252877 TI - [Relationships of Aulacophora beetles feeding behavior with cucurbitacin types in host crops]. AB - Aulacophora fermoralis chinensis and Aulacophora cattigarensis are the two related Aulacophora beetles, but their host selectivity and feeding behavior are significantly different. A. fernzoralis chinensis usually feeds upon Cucumis sativus and Cucurbita moschata, but never upon Luffa acutangula, Momordica charantia and Citrullas lanatus. Its feeding behavior on hosts is to snip a circular trench on their leaves, and then, to feed the leaf tissues isolated by the trench. On the contrary, A. cattigarensis only feeds L. acutangula, and its trenching behavior rarely occurs. This study showed that the host selectivity and feeding behavior of the two beetles were significantly correlated with the types of cucurbitacin biosynthesized by host melon crops. C. sativus and C. moschata might be induced to produce cucurbitacin I by A. fermoralis chinensis feeding, and this compound was responsible for feeding deterrent on A. fermoralis chinensis. Therefore, the feeding behavior of A. fermoralis chinensis was to block the translocation of cucurbitacin I to feeding sites. M. charantia and C. lanatus contained deterrent cucurbitacin D, and thus, both beetles never fed upon them, while L. acutangula contained stimulants cucurbitacin B and E, and made A. cattigarensis directly feed upon it without trenching. The results suggested that the taste responses of insects on specific allelochemicals from plants may play an important role in host selectivity and feeding behavior. PMID- 16252879 TI - [Hydrochemical characteristics of three kinds of wetland in Gongbiela Basin]. AB - The study on the hydrochemical characteristics of three representative kinds of wetland in Gongbiela Basin showed that in the water of test wetlands, HCO3 - was the dominant anion, accounting for 81.91% - 85.46% of total anions, and Ca2+ was the dominant cation, accounting for 56.80% - 69.32% of total cations. The hydrochemical type belonged to that of bicarbonate calcium. In the three kinds of wetland, water pH ranged from 6.2 to 7.1, mineralization degree ranged from 112.56 to 461.23 mg x L(-1), and hardness ranged from 14.31 to 148.53 mg x L(-1). On the whole, the water quality of the wetlands met the grade 1 and grade 2 national environmental water quality standards, but the Fe and Mn contents exceeding the standards influenced the water resource quality of this area. The spatial and temporal changes of hydochemical characteristics of the wetlands and the trace element contents in the water were also discussed and analyzed. PMID- 16252880 TI - [Zooplankton in north branch waters of Changiiang Estuary]. AB - Based on the investigation data during the high-water (July, 2003) and low-water (January, 2004) periods, a causal analysis was made on the variation of zooplankton distribution in the north branch waters of the Changjiang Estuary. The results showed that in high-water period, the average of zooplankton biomass was 234.38 mg x m(-3), being 141.35 mg x m(-3) in flood tide and 327.40 mg x m( 3) in ebb tide, while in low-water period, it was 188.81 mg x m(-3), being 184.69 mg x m(-3) in flood tide and 192.93 mg x m(-3) in ebb tide. The biomass increased from the east to the west in flood tide, but a contrary trend was observed in ebb tide. The species number did not change obviously both in flood tide and in ebb tide. The value of diversity index (H') was higher in flood tide than in ebb tide. In high-water period, the biomass near the north shore was higher than that near the south shore, but it was contrary in the ebb tide. The difference between the waters of two shores was not obvious in low-water period as in high-water period, though the trend of biomass variation was similar. The variation of zooplankton distribution in the north branch waters of the Changjiang Estuary had a close relation with the seasonal changes of zooplankton biomass outside the Changjiang Estuary and the tide, but not significantly related with the Changjiang runoff water. Coriolis force accounted for the difference of zooplankton biomass in the waters of two shores via tide movement. PMID- 16252881 TI - [Recruitment dynamics of bloom-forming cyanobacteria in Meiliang Bay of Taihu Lake]. AB - A collection trap was installed on the sediments in Meiliang Bay of Taihu Lake during the period from March to June to investigate the recruitment of phytoplankton, and cyanobacteria in particular. The algal abundance in the trap was monitored, and compared with that in water column and surface sediments. The results showed that the algal recruitment was related to the temperature, irradiance, DO content, and oxidation-reduction potential (ORP) in surface sediment. After over-wintering on the sediments, the colonies of cyanobacteria experienced a prior developmental process before re-invasion of the water column. During the investigation period, the recruitment rates of chlorophyll a and b and PC were 59.84 %, 76.83 % and 466.98%, and the recruitment amounts accounted for 7.18%, 3.71% and 9.33% of their maximum pelagic counterparts in water column, respectively. These results showed that shallow lake was the important seed bank to the pelagic populations, and water bloom was formed as the result of the recruitment and biomass accumulation of cyanobacteria. PMID- 16252882 TI - [Accumulation of non-point source pollutants in ditch wetland and their uptake and purification by plants]. AB - The study on the vertical and horizontal distribution of organic matters and total nitrogen (TN) in the sediment of ditch wetland naturally grown with reed (Phragmites communis) and wild rice (Zizania latifolia) showed that the sediment below 40 cm depth had a significant effectiveness in retaining and accumulating organic matters and TN, but in its surface layer, this effectiveness varied largely with seasons, and the maximum was more than twofold of the minimum. TN was highly correlated with organic matters, the correlation coefficient being 0.9876 in reed wetland and 0.9335 in wild rice wetland, and in water phase, it was positively related to NH4+ -N and NO3-N, indicating that the main composition of TN was organic N, and the mineralization of organic N was the sources of inorganic N. The harvest of reed in each autumn could take away 818 kg x hm(-2) of N and 103.6 kg x hm(-2) of P, and that of wild rice could take away 131 kg x hm(-2) of N and 28.9 kg x hm(-2) of P. Zizania caduci flora had a high assimilation ability of nutrients. Its cultivation in ditch wetland to replace wild helophytes would be a good approach to attain higher absorbing ability of N and P, and to resolve the secondary pollution problem of emerged plants, because farmers could harvest it voluntarily. PMID- 16252883 TI - [Impacts of petroleum-containing wastewater irrigation on microbial population and enzyme activities in paddy soil of Shenfu irrigation area]. AB - The study showed that the upper reaches of main petroleum-containing wastewater irrigation channels had the highest accumulation and distribution of total petroleum hydrocarbon (TPH) (5 213.37 mg x kg(-1) dry soil),and the CFU of soil bacteria and fungi was increased with increasing soil TPH concentration, the correlation coefficient being 0.928 (P < 0.001) and 0.772 (P < 0.05), respectively. The activities of soil dehydrogenase, catalase and polyphenoloxidase had a significantly positive correlation with soil TPH concentration, their correlation coefficient being 0.974 (P < 0.001), 0.957 (P < 0.001) and 0.886 (P < 0.001), respectively, while soil urease activity showed a significantly negative correlation (P = 0.002), which could be used as the most sensitive indicator of petroleum contamination. The substrate-induced respiration (SIR) of polluted soil was significantly correlated with soil TPH concentration (P < 0.001), dehydrogenase activity (P < 0.001), and heterotrophic bacterial CFU (P = 0.006). PMID- 16252885 TI - [Effects of urbanization on landscape pattern of Beijing]. AB - Landscape pattern of Beijing urban areas within the six-ringed road was classified based on 1997 and 2002 remote sensing data. The indices from landscape level were computed using Fragstats 3.3 to study the landscape pattern change of Beijing urban areas from 1997 to 2002. The results showed that the number of patches increased by 106.6%, the mean patch area decreased by 51.6%, the splitting index increased by 94.3%, and the landscape diversity index increased by 17.8%. The landscape pattern tended to be fragmentized and diversified in those five years. The reason of this change was the increased need for construction land due to the growing population and the improvement of living standard. Landscape pattern change of Beijing might influence the changes of substance flow and energy flow. It also provided some fundamental landscape scale data and some suggestions for development and planning of Beijing city. PMID- 16252884 TI - [Effects of environmental factors on cd biotoxicity and phytochelatins production in Triticum aestivum]. AB - In this paper, a solution culture experiment was conducted to study the effects of environmental factors on Cd biotoxicity and phytochelatins (PCs) production in wheat. The results showed that Cd stress had significant inhibitory effects on wheat growth and PCs overproduction. The Cd biotoxicity and Cd uptake by wheat were affected in varying degrees by soil pH, Ca and S, and the levels of PCs production in root were consistent with the changes of Cd biotoxicity. Furthermore, the Cd biotoxicity was decreased with increasing P supply, coinciding with the decrease of PCs level in root. Mg had no obvious effect on both Cd biotoxicity and PCs level in root. The present results further confirmed that the induced PCs production level by Cd was related to Cd biotoxicity in plant, suggesting that PCs could be a promising biomarker for estimating Cd phytotoxicity. PMID- 16252886 TI - [Ecological footprint and available ecological capacity in Chongqing region]. AB - Based on the statistical data of Chongqing, the ecological footprint of Chongqing was calculated in this paper. The results showed that the per capita ecological footprint was 1.653566 hm2, per capita ecological capacity was 0.280393 hm2, and ecological surplus of deficit was 1.373173 hm2. The per capita ecological footprint was 0.5335 hm2 (47.64%) higher but the per capita ecological capacity was 0.5196 hm2 (64.95%) lower, and the ecological surplus of deficit was about 3.43 times of the average national level. These results showed that the ecological footprint of Chongqing was beyond the available ecological capacity, and its social and economic development was not sustainable. The strategies on reducing ecological deficit in this region, such as reducing ecosystem population, increasing public finance income, and controlling environmental pollution, were also put forward. PMID- 16252887 TI - [Animal immunocompetence and its effect on population regulation and life history trade-off]. AB - Immunocompetence is a common conception in biomedical and animal sciences, and refers to the ability of organisms against diseases. In recent years, a new field, ecological immunology, has been received much attention, which deals with the variation of immunocompetence in wildlife by immune methods. This paper introduced the conception of immunocompetence, the factors affecting its performance, and its roles in regulating animal population dynamics. Some hypotheses on population regulation in small mammals, such as immunocompetence handicap hypothesis, immunocompetence selection hypothesis and winter immune enhancement hypothesis were summarized. The relationships among energy metabolism, endocrine controlling mechanism and immune system were introduced, and the recent progress on immunocompetence and life history trade-off was analyzed. The trade-off between survival and immunocompetence, as well as between reproduction and immunocompetence were discussed, and some developing perspectives and new problems in ecological immunology were proposed. PMID- 16252888 TI - [Bacterial flora composition and its dynamics in tidal-flat Sinonovacula constricta aquaculture area]. AB - In this study, the bacteria from the mud in tidal-flat Sinonovacula constricta aquaculture area were isolated each month from March to December, 2002, and the temporal and spatial distribution of heterotrophic bacteria, ammonifying bacteria, denitrifying bacteria, and sulphate reducing bacteria were analyzed. The results showed that all the 515 isolated bacteria mainly belonged to 1 family and 13 genera. The bacterial flora in different layers of the mud was almost consistent, while the composition was different. The predominant genera were Clostridium, Bacillus, Corynebacterium, Photobacterium, and some Enterobacteriaceae. The number of heterotrophic bacteria in the surface layer and the bottom fluctuated in 7.6 x 10(3) cfu x g(-1) - 2.0 x 10(5) and 1.6 x 10(3) - 1.0 x 10(5) cfu x g(-1), ammonifying bacteria fluctuated in 1.5 x 10(6) - 9.0 x 10(7) and 9.0 x 10(5) - 1.0 x 10(7) cfu x g(-1), denitrifying bacteria fluctuated in 9.0 x 10(3) - 4.0 x 10(6) and 5.0 x 10(2) -1.9 x 10(6) cfu x g(-1), and sulphate reducing bacteria fluctuated in 5.0 x 10(4) - 5.0 x 10(6) and 1.9 x 10(4) - 2.0 x 10(6) cfu x g(-1), respectively. The detection rates of ammonifying bacteria, denitrifying bacteria and sulphate reducing bacteria in the mud were all 100%, and these bacteria increased significantly in the second half of the year, indicating that the environment of the Sinonovacula constricta aquaculture area was deteriorated due to the accumulation of NH3, nitrite and H2S, and it is important to regulate the breed capacity and redistribute the breeding environment. PMID- 16252889 TI - Frontotemporal dementia--part III. Clinical diagnosis and treatment. AB - The authors report a comprehensive publication consisting of three parts going into the details of history, prevalence, clinical forms, differential diagnosis, genetics, molecular pathomechanism, and pathology, clinical diagnosis and treatment of frontotemporal dementia (FTD). The third part of the present review focuses on the clinical diagnosis and treatment of FTD. The diagnosis of FTD is problematic even today. Mental status, psychometric testing as well as imaging studies such as PET and SPECT, and laboratory examinations may be helpful in the diagnosis. Unfortunately, bedside tests are generally insensitive to mild executive deficits. Most of FTDs do not have characteristic laboratory abnormalities or brain atrophy sufficient to set up the diagnosis; these only allow to rule out other disorders and assume the diagnosis of FTD. An effective treatment for FTD is still to be established. The improvement of serotonin metabolism has been proposed as a biological treatment. Recent studies suggest that bromocriptine may improve selective frontal symptoms, but this and the efficacy of other dopaminergic drugs need further evaluation. Drugs that prevent the expression or accumulation of tau seem to be the most promising causal approach. In aphasia behavioral therapy may be attempted. In addition, caregiver management is essential, because as with those of Alzheimer's dementia patients, they also carry a significant psychosocial stress. PMID- 16252890 TI - [Regulation of water transport in brain oedema]. AB - The study gives an overview on the regulation of cerebral water content and of brain volume. The molecular mechanisms of the development and resolution of various oedema forms are discussed in detail. The physiological and pathophysiological role of the recently discovered molecular water channel proteins aquaporin-1 (AQP1) and aquaporin-4 (AQP4) as well as the importance of central neuroendocrine regulation by vasopressin and atriopeptin are reviewed based on the relevant literature and personal studies. Quantitative water maps based on the combination of multicompartment-T2, diffusion weighted MRI and T1 studies have proven to be powerful tools for studying new drugs against brain oedema brought about by various neuropathological conditions and for testing their efficacy both in animal experimental and clinical conditions. Non-peptide vasopressin antagonists, atriopeptin agonists and drugs targeting AQP4 are potential new families of oedema-decreasing drugs. PMID- 16252891 TI - [Ischemic preconditioning in the brain]. AB - Stroke-related death is the third most common cause of mortality in Hungary after cardiovascular diseases and cancer. In addition to the unfavourable mortality figures, the consequent development of neurological and psychiatric disorders in stroke patients imposes an enormous burden on the health care system and on the families. Numerous studies are being conducted world wide on the prevention of stroke and other cerebrovascular disorders like chronic hypoperfusion, as well as on acute stroke management and patient rehabilitation issues. As a result, our understanding of the mechanism of hypoxic brain damage steadily increased over the past years and decades which brought along promising achievements both in the field of stroke prevention and therapy. However, these broad-spectrum approaches also made clear that preventing neuronal death and thus reducing neurological damage are complex tasks that cannot be successfully resolved by targeting single mechanisms. Therefore, the development of alternative new drugs and clinically applicable complex neuroprotective strategies is warranted. One of the most promising approaches is to create ischemic tolerance in the brain by using pharmacological preconditioning paradigms. These drugs trigger similar events to those initiated by brief ischemic insults that later can make the cerebral tissue resistant to subsequent otherwise lethal stress (ischemic preconditioning). PMID- 16252892 TI - Neurobehavioral assessment in the neonatal period--the impact of Ferenc Katona. AB - BACKGROUND AND PURPOSE: Identifying and treating CNS-injured infants has always been Ferenc Katona's main goal. Early on, he showed the importance of "elementary motor patterns" to development, and their application in treatment of early brain injury. We report on a neonatal neurobehavioral assessment derived from Katona that has concurrent and predictive validity in the NICU population. It evaluates early behavioral capabilities and dysfunctions in areas often disrupted by CNS injury, such as attention, motor organization, and autoregulation; provides a valuable approach for assessing recovery from CNS injury; and predicts later mental and motor performance. METHODS: A population-based NICU sample (n=3226) studied from 1989-2004 had CNS injury ranging from none to severe. Our neurobehavioral assessment is a categorical clinical evaluation suitable for small, sick, high-risk neonates that is composed of visual, auditory, head control, tone, state, and elicited active motor behaviors designed to challenge the infant. Infants are tested at hospital discharge and one month post term. A series of tasks are administered enabling a trained examiner to make judgments of abnormality on over 20 categories of behavior. RESULTS: Maximum likelihood ordered logistic regression and multiple regression specified the relation of abnormality in neurobehavioral performance to concurrent estimates of CNS injury and to later cognitive and motor outcome. All behavioral categories were significantly associated with CNS abnormality at one or both ages (p's<0.03 0.0001). Multiple regression showed a significant linear relationship (p<0.0001) of CNS injury to number of neurobehavioral abnormalities, which decreased with decreasing injury and across age. CONCLUSIONS: Katona's strong contributions form the basis of neurobehavioral assessment that can detect early abnormalities and help design early interventions. He is unique in his understanding of the neural mechanisms underlying early CNS injury and its behavioral consequences. His concepts allowed us to devise a valid newborn neurobehavioral assessment to evaluate early behavioral capabilities frequently disrupted by CNS injury. PMID- 16252893 TI - [Evolutionary pressure in the development of the unconscious-conscious duality in young children]. PMID- 16252894 TI - Bladder rehabilitation in neurogenic and non-neurogenic detrusor dysfunction with intravesical electrotherapy. PMID- 16252896 TI - At capacity and beyond. Ideas such as 'surge' hospitals are getting a more careful look as healthcare wrestles with planning for large-scale disasters. AB - "Surge" hospitals, which can handle an influx of patients during a large-scale disaster, are being considered by some who think the healthcare industry's plans for such catastrophes need work, says Jim Bentley, left, senior vice president for strategic policy planning of the American Hospital Association. PMID- 16252895 TI - Complex elementary movements of humans--the anthropological approach of Ferenc Katona. PMID- 16252897 TI - Double whammy. Katrina evacuees in Houston flee Rita. PMID- 16252898 TI - An up-Hill battle. Getting money to disaster areas proves challenging. PMID- 16252899 TI - Surgeries' quality questioned. PMID- 16252900 TI - Plans extol community benefits. PMID- 16252901 TI - Traditional plans preferred: poll. PMID- 16252902 TI - Feds subpoena Iasis. A whistle-blower lawsuit may be behind investigation. PMID- 16252903 TI - The real lessons of New Orleans. Along with the blame, how about some perspective and solutions? PMID- 16252904 TI - Creativity rules. Design winners use architectural themes successfully. PMID- 16252905 TI - Opening doors. Harvard Medical School's Joan Reede is working to narrow the racial gaps in healthcare leadership. PMID- 16252906 TI - Bisabolane-type sesquiterpenes: liginvolones A-D from ligusticum involucratum. AB - Four new bisabolane-type sesquiterpenes, liginvolones A-D (1-4), a new coumarin, 4'-octanoyloxyosthenol (5), and 20 known constituents were isolated from Ligusticum involucratum. Their structures were elucidated on the basis of spectroscopic analyses and chemical methods. PMID- 16252907 TI - Antineoplastic agents. 509: synthesis of fluorcombstatin phosphate and related 3 halostilbenes(1). AB - The present SAR study of combretastatin A-3 (3a) focused on replacement of the 3 hydroxyl group by a series of halogens. That approach with Z-stilbenes resulted in greatly enhanced (>10-100-fold) cancer cell growth inhibition against a panel of human cancer cell lines and the murine P388 lymphocytic leukemia cell line. Synthesis of the 3-fluoro-Z-stilbene designated fluorcombstatin (11a) and its potassium 3'-O-phosphate derivative (16c) by the route 7 --> 8a --> 11a --> 14 - > 16c illustrates the general synthetic pathway. The 3'-O-phosphoric acid ester (15) of 3-bromo-Z-stilbene 13a was also converted to representative cation salts to evaluate the potential for improved aqueous solubility, and the potassium salt (16 mg/mL in water) proved most useful. The fluoro (11a), chloro (12a), and bromo (13a) halocombstatins were nearly equivalent to combretastatin A-4 (1a) as inhibitors of tubulin polymerization and of the binding of colchicine to tubulin. The tubulin binding in cell-free systems was also retained in human umbilical vein endothelial cells. All three halocombstatins retained the powerful human cancer cell line inhibitory activity of combretastatin A-4 (1a) and proved superior to combretastatin A-3 (3a). In addition, the halocombstatins targeted Gram-positive bacteria and Cryptococcus neoformans. PMID- 16252908 TI - Effect of benzylic oxygen on the antioxidant activity of phenolic lignans. AB - It has been clarified in the present investigation that a high degree of oxidation at the benzylic position of phenolic lignans bearing a 4-hydroxy-3 methoxybenzyl group reduces their antioxidant activity and that the antioxidant activity of the bis(4-hydroxy-3-methoxybenzyl)tetrahydrofuran lignan 2 is higher than that of the corresponding gamma-butyrolactone lignan 1. This was demonstrated by comparing the antioxidant activities of compounds 1 and 2 with those of the (benzyl)(hydroxybenzyl)tetrahydrofurans 3 and 4, the bis(hydroxybenzyl)tetrahydrofurans 7 and 8, the (benzoyl)(benzyl)tetrahydrofuran 6, and the dibenzoyltetrahydrofuran 9. The activity level of compound 2 was approximately the same potency as that of the tetrahydronaphthalene tetrahydrofuran 5. These compounds possess either a 4-hydroxy-3-methoxybenzyl group or a 4-hydroxy-3-methoxybenzoyl group as the benzyl or benzoyl group. An examination of radical scavenging activity showed differences of activity between diastereomers. To make this comparison possible, compounds 1-9 were synthesized using new synthetic routes for several of these lignans. In this investigation, stereoisomers of the (benzyl)(hydroxybenzyl)tetrahydrofurans 3 and 4 and liovils 7 and 8 were synthesized for the first time. PMID- 16252909 TI - Labdane diterpenes from Aster spathulifolius and their cytotoxic effects on human cancer cell lines. AB - Three new labdane diterpenes (1-3), together with eight known diterpenoids, were isolated from a methanol extract of the aerial parts of Aster spathulifolius. The structures of 1-3 were determined as (13R)-labda-7,14-diene 13-O-beta-d-(4'-O acetyl)fucopyranoside (1), (13R)-labda-7,14-diene 13-O-beta-d-(3'-O acetyl)fucopyranoside (2), and (13R)-labda-14(15)-en-8,13-diol 13-O-beta-d fucopyranoside (3), on the basis of spectroscopic and chemical methods. Compounds 1, 2, and four of the known compounds exhibited generally nonspecific cytotoxicity against human A549, SK-OV-3, SK-MEL-2, XF498, and HCT15 tumor cells. PMID- 16252910 TI - Cytotoxic phenylpropanoid glycosides from the stems of Smilax china. AB - Bioassay-guided fractionation of an ethanol extract of Smilax china led to the isolation of nine phenylpropanoids including six new compounds, smilasides A-F (1 6), and three known phenylpropanoids, smiglaside E, heloniosides B, and 2',6' diacetyl-3,6-diferuloylsucrose. Structural elucidation of isolates 1-6 was based on spectroscopic data analysis. These new phenylpropanoids were evaluated against several human tumor cell lines. PMID- 16252911 TI - Chemical and biological properties of an arabinogalactan from Phyllanthusniruri. AB - Phyllanthus niruri is a well-known herb widely used medicinally in Asia, Africa, and South America. Aqueous extraction of the intact plant provided an acidic arabinogalactan, which was characterized chemically, and its effects on peritoneal macrophage activation were determined. Methylation analyses and (13)C NMR spectroscopy showed it to have a complex structure with a (1-->4)-linked beta Galp main chain, substituted by rhamnose, galacturonic acid, arabinose, xylose, galactose, and glucose-containing side chains, with nonreducing end-units of arabinofuranose, xylopyranose, galactopyranose, and glucopyranose. In immunological studies, the arabinogalactan stimulated superoxide anion production, when tested using peritoneal macrophages of mice, but did not interfere with the nitric oxide pathway. Thus, traditional aqueous extraction methods, such as decoction and infusion, provide a major polysaccharide, which stimulates an intense biological response in macrophages: this could represent an interesting approach in phytotherapeutic treatments. PMID- 16252912 TI - Investigation of brominated tryptophan alkaloids from two thorectidae sponges: Thorectandra and Smenospongia. AB - Chemical investigation of an NCI-DTP collection of Thorectandra sp. and a UCSC collection of Smenospongia sp. yielded six new brominated tryptophan derivatives: 6-bromo-1'-hydroxy-1',8-dihydroaplysinopsin (4), 6-bromo-1'-methoxy-1',8 dihydroaplysinopsin (5), 6-bromo-1'-ethoxy-1',8-dihydroaplysinopsin (6), (-)-5 bromo-N,N-dimethyltryptophan (7), (+)-5-bromohypaphorine (8), and 6-bromo-1H indole-3-carboxylic acid methyl ester (11). Additionally, the known compounds aplysinopsin (1), 1',8-dihydroaplysinopsin (2), 6-bromo-1',8-dihydroaplysinopsin (3), (1H-indole-3-yl)acetic acid (9), and (6-bromo-1H-indol-3-yl)acetic acid methyl ester (10) were also encountered. The structures of 4-8 and 11 were confirmed on the basis of analysis of (1)H and (13)C (1D and 2D) NMR data as well as comparison to known compounds. Compounds 1, 3-8, 10, and 11 were found to inhibit the growth of Staphylococcus epidermidis with either weak or moderate MICs. PMID- 16252913 TI - Steroidal saponins from Smilax medica and their antifungal activity. AB - Three new steroidal saponins (1-3) were isolated from the roots of Smilax medica, together with the known disporoside A (4). The structures of the new compounds were elucidated mainly by extensive spectroscopic analysis (1D and 2D NMR, FABMS, and HRESIMS). Compounds 1, 2, and 4 demonstrated weak antifungal activity against the human pathogenic yeasts Candida albicans, C. glabrata, and C.tropicalis, with MIC values between 12.5 and 50 microg/mL. PMID- 16252914 TI - Cytotoxic principles from the formosan milkweed, Asclepias curassavica. AB - A series of cardenolides and related compounds have been isolated from the aerial parts and roots of the ornamental milkweed, Asclepias curassavica. Their structures were determined by spectroscopic and chemical methods. Among them, three derivatives of calactinic acid methyl ester (13-15), 19-nor-16 alpha acetoxy-10 beta-hydroxyasclepin (16), 20 beta,21-dihydroxypregna-4,6-dien-3-one (19), and 3,4-seco-urs-20(30)-en-3-oic acid (22) are new compounds. The relative configuration of calactinic acid methyl ester (12) has been confirmed by X-ray diffraction analysis on its derivative 13. Most of the cardenolides obtained showed pronounced cytotoxicity against four cancer cell lines (IC(50) 0.01 to 2.0 microg/mL). PMID- 16252915 TI - Rapid extract dereplication using HPLC-SPE-NMR: analysis of isoflavonoids from Smirnowia iranica. AB - A novel hyphenated technique, HPLC-SPE-NMR, was used for accelerated identification of isoflavonoids from the roots of Smirnowia iranica. The extract constituents eluted from a HPLC column were automatically trapped on solid-phase extraction (SPE) cartridges, and NMR spectra were acquired with concentrated solutions after solvent change. The structures of 10 new isoflavonoids (1, 4, 5, 7-10, 12, 13, 16) and of seven previously described constituents (2, 3, 6, 11, 14, 15, 17) were elucidated from NMR spectra acquired in the HPLC-SPE-NMR mode. Multiple peak trapping on the same SPE cartridge increased analyte amounts and provided access to 2D NMR data. It was demonstrated that linear accumulation of material is possible in up to seven repeated trapping steps. The use of HPLC-SPE NMR speeded up dereplication of the S. iranica extract considerably by providing detailed information about the constituents of a complex, essentially crude extract prior to their preparative-scale isolation or extract pre-fractionation, and the information obtained could be used to direct preparative isolation work. In connection with structure elucidation of isoflavonoids containing O-methylated 1,2,3-benzenetriol moieties as the B-ring, O-methylation-induced changes of chemical shifts of aromatic hydrogens were found to depend on the conformation of the resulting methoxy group, i.e., on the number of its ortho substituents. The recognized regularities will be useful in structure determination of partially O methylated polyphenols based on 1D (1)H NMR spectra obtainable from HPLC-SPE-NMR experiments, diminishing dependence on 2D NMR data and (13)C NMR chemical shifts. PMID- 16252916 TI - Nematicidal epipolysulfanyldioxopiperazines from Gliocladium roseum. AB - Five new verticillin-type epipolysulfanyldioxopiperazines, gliocladine A (1), B (2), C (3), D (4), and E (5), were isolated from wheat solid-substrate fermentation of Gliocladium roseum 1A, along with four known compounds, verticillin A (6), 11'-deoxyverticillin A (7), Sch52900 (8), and Sch52901 (9). Their structures were elucidated by extensive 1D and 2D NMR studies, MS, and chemical transformations. In vitro immersion tests showed that all nine compounds exhibited antinematodal activity against Caenorhabditis elegans and Panagrellus redivivus. The monomeric epipolysulfanydioxopiperazines (3-5), with the indole moiety, were found to be less active than the dimeric compounds (1, 2, 6-8). PMID- 16252918 TI - Caucanolides A-F, unusual antiplasmodial constituents from a colombian collection of the gorgonian coral Pseudopterogorgia bipinnata. AB - Six new diterpenoids, caucanolides A-F (1-6), have been isolated from extracts of the gorgonian octocoral Pseudopterogorgia bipinnata collected near the Colombian Southwestern Caribbean Sea. The structures of 1-6 were elucidated by comprehensive analysis of spectroscopic data. The caucanolides showed in vitro antiplasmodial activity against the malaria parasite, Plasmodium falciparum. In addition to possessing structures based on novel carbon skeletons, one of these metabolites, caucanolide B (2), constitutes the only example from nature of a secondary metabolite possessing the N(1),N(1)-dimethyl-N(2)-acylformamidine functionality. PMID- 16252917 TI - Selective cyclooxygenase-2 inhibitors from Calophyllum membranaceum. AB - Chemical investigation of the anti-inflammatory Chinese folk medicine Calophyllum membranaceum has resulted in the isolation and characterization of three new xanthones (1-3), one new biphenyl C-glycoside (4), and one new phenylethanoid glycoside (5) along with 17 known compounds. Their structures were characterized on the basis of spectroscopic and chemical methods. Two xanthones, 2,6-dihydroxy 1,7-dimethoxyxanthone (1) and 3,4-dihydroxyxanthone, were found to exhibit selective inhibitory activity against cyclooxygenase-2 (IC(50)=2.99 and 1.80 microM) in vitro. PMID- 16252919 TI - Glycolipids from sponges. Part 16.(1) discoside, a rare myo-inositol-containing glycolipid from the caribbean sponge Discodermia dissoluta. AB - Discoside (1a), a glycolipid composed of 4,6-O-diacylated mannose attached to the 2-hydroxyl group of a myo-inositol unit, was isolated as a mixture of homologues from the marine sponge Discodermia dissoluta. The complete stereostructure of this new glycolipid was solved by interpretation of mass spectrometric and NMR data and CD analysis of degradation products. PMID- 16252920 TI - Phenolic and triterpene glycosides from the stems of Ilex litseaefolia. AB - Chemical investigation on the stems of Ilex litseaefolia afforded four new phenolic glycosides, litseaefolosides A-D (1-4), and two new triterpene glycosides, spathodic acid 28-O-beta-d-glucopyranoside (5) and (20S)-niga ichigoside F1 (6), along with 28 known compounds. The structures of 1-6 were determined on the basis of chemical and spectroscopic evidence. Litseaefoloside C (3) showed inhibitory activities in vitro for alpha-glucosidase and lipase with IC(50) values of 34.0 and 0.31 microg/mL, respectively. PMID- 16252922 TI - Phrygiasterol, a cytotoxic cyclopropane-containing polyhydroxysteroid, and related compounds from the pacific starfish Hippasteria phrygiana. AB - The new cyclopropane-containing steroid phrygiasterol (1) and steroid glycoside phrygioside B (2), along with previously known borealoside C (2a) and (20R,24S) 5alpha-cholesta-3beta,6alpha,8,15alpha,24-pentaol (3), have been isolated from the Pacific starfish Hippasteria phrygiana. On the basis of spectroscopic analyses, using 1D and 2D NMR techniques and some chemical transformations, the structures of 1 and 2 have been established as (20R,24R,25R)-24,26-cyclo-5alpha cholesta-3beta,6alpha,8,15alpha,16beta,27-hexaol (1) and the sodium salt of (20R,24S)-24-O-(3-O-methyl-4-O-sulfate-beta-d-xylopyranosyl)-5alpha-cholesta 3beta,6alpha,8,15alpha,24-pentaol (2), respectively. Compound 1 inhibited the growth of Ehrlich carcinoma cells with an IC(50) of 50 microg/mL, whereas 2 induced apoptosis of the same cells (EC(50)=70 microg/mL) and inhibited Ca(2+) influx into mouse spleenocytes (EC(50)=20microg/mL). PMID- 16252923 TI - Anti-inflammatory acylphloroglucinol derivatives from Hops (Humulus lupulus). AB - The polyphenol-enriched fraction of an ethanolic hops extract (Humulus lupulus) was separated to provide four acylphloroglucinol-glucopyranosides (1-4). 1-(2 Methylpropanoyl)phloroglucinol-glucopyranoside 1 has been isolated from hops before, whereas 1-(2-methylbutyryl)phloroglucinol-glucopyranoside 2, known as multifidol glucoside, and 1-(3-methylbutyryl)phloroglucinol-glucopyranoside 3 were found in hops for the first time. 5-(2-Methylpropanoyl)phloroglucinol glucopyranoside 4 was identified as a new natural product. The compounds were tested for inhibition of COX-1 activity. The aglycon 5, obtained by acid hydrolysis of 1, was equally effective as phloroglucinol, with an IC(50) of 3.8 microM. The inhibitory potential of the glucosides was 1>2>3 and decreased with increasing length of the acyl side chain. Compound 4 was about 2.5-fold less active than 1 (IC(50): 23.7 and 58.7 microM, respectively). PMID- 16252921 TI - Retinoylserine and retinoylalanine, natural products of the moth Trichoplusia ni. AB - Insect cells convert vitamin A into a number of retinoids that are evolutionarily conserved with those of mammalian cells. However, insect cells also produce additional natural retinoids. Namely, two retinoic acid peptides, N-trans retinoylserine (1) and N-trans-retinoylalanine (2), have been isolated from a cell line of the common cabbage looper, Trichoplusia ni. These are the first examples of naturally occurring retinoic acid linked to amino acids through an amide bond; the amino acid moieties are depicted in the more common l configuration, although the absolute configuration was not determined due to the minuscule sample amount. PMID- 16252925 TI - Halogenated helianane derivatives from the sponge Spirastrella hartmani. AB - Two new halogenated derivatives (1 and 2) of helianane (3) were isolated from the 2-propanol extract of the sponge Spirastrella hartmani. The structures of the new derivatives were determined on the basis of detailed spectroscopic analysis, including (+)-HREIMS and 1D and 2D NMR. Compound 1 showed in vitro cytotoxicity against the human tumor cell lines A549, HT29, and MDA-MB-231. PMID- 16252924 TI - Cytotoxic furostanol saponins and a megastigmane glucoside from tribulus parvispinus. AB - Two new furostanol saponins, (25R)-26-O-beta-D-glucopyranosyl-5alpha-furostan 2alpha,3beta,22alpha,26-tetraol 3-O-{beta-D-galactopyranosyl-(1-->2)-O-[beta-D xylopyranosyl-(1-->3)]-O-beta-D-glucopyranosyl-(1-->4)-beta-D-galactopyranoside} (1) and (25R)-26-O-beta-D-glucopyranosyl-5alpha-furostan-3beta,22alpha,26-triol 3 O-{beta-D-galactopyranosyl-(1-->2)-O-[beta-D-xylopyranosyl-(1-->3)]-O-beta-D glucopyranosyl-(1-->4)-beta-D-galactopyranoside} (2), and their O-methyl derivatives (3 and 4), and a new megastigmane glucoside, (6S,7E,9xi)-6,9,10 trihydroxy-4,7-megastigmadien-3-one 10-O-beta-D-glucopyranoside (6), along with one known spirostanol saponin, gitonin (5), and four known megastigmane glucosides were isolated from the aerial parts of Tribulus parvispinus. Their structures were established by detailed spectroscopic analysis. The cytotoxic activities of 1-6 against U937, MCF7, and HepG2 cells were evaluated. Compounds 2 (IC(50) 0.5 microM) and 5 (IC(50) 0.1 microM) showed the highest activity against U937 cells. PMID- 16252926 TI - Alkaloids from amphibian skin: a tabulation of over eight-hundred compounds. AB - A diverse array of biologically active, lipid-soluble alkaloids have been discovered in amphibian skin. Such alkaloids include the following: the steroidal samandarines from salamanders, the batrachotoxins, histrionicotoxins, gephyrotoxins, and epibatidine from neotropical poison frogs (Dendrobatidae), the pumiliotoxins, allopumiliotoxins, homopumiliotoxins, and decahydroquinolines from certain genera of anurans from four families (Dendrobatidae, Mantellidae, Bufonidae, and Myobatrachidae), a variety of izidines (pyrrolizidines, indolizidines, quinolizidines, lehmizidines), pyrrolidines, piperidines, various tricyclics (related in structures to the coccinellines), and spiropyrrolizidines from the first three of these four families, the pseudophrynamines from one genus of Australian frogs, and a variety of unclassified alkaloids as yet of undetermined structure. With the exception of the samandarines and the pseudophrynamines, all alkaloids appear to be derived from dietary sources. Although only a few of the over 800 amphibian skin alkaloids have been detected in arthropods, putative arthropod sources for the batrachotoxins and coccinelline like tricyclics (beetles), the pumiliotoxins (ants, mites), the decahydroquinolines, izidines, pyrrolidines, and piperidines (ants), and the spiropyrrolizidines (millipedes) have been discovered. Ants are likely sources for histrionicotoxins, lehmizidines, and tricyclic gephyrotoxins. Epibatidines represent an important alkaloid class without a putative dietary source. The structures for many of these alkaloids have been rigorously established, while the structures of others represent tentative proposals, based only on mass spectral and FTIR spectral data, along with analogies to structures of well defined alkaloids. PMID- 16252927 TI - Diaper dermatitis: a review and brief survey of eruptions of the diaper area. AB - Eruptions in the diaper area are the most common dermatologic problem in infancy. Such eruptions can be subdivided into primary diaper dermatitis, an acute inflammation of the skin in the diaper area with an ill-defined and multifactorial etiology, and secondary diaper dermatitis, a term which encompasses eruptions in the diaper area with defined etiologies. The most important factors in the development of primary diaper dermatitis are: (i) water/moisture, (ii) friction, (iii) urine, (iv) feces, and (v) microorganisms (sometimes). Possible treatments include minimizing diaper use and using disposable diapers, barrier creams, mild topical cortisones, and antifungal agents. A variety of other inflammatory and infectious processes can occur in the diaper area and an awareness of these secondary types of diaper dermatitis aids in the accurate diagnosis and treatment of patients. PMID- 16252928 TI - Optimal management of severe plaque form of psoriasis. AB - Psoriasis is a chronic, inflammatory, hyperproliferative skin disease that affects 1-2% of the general population in the UK and US. Plaque psoriasis is the most common form, accounting for approximately 90% of cases. The disease is usually chronic and persistent, although up to 50% of patients may enter spontaneous remission for varying periods of time. There is no cure for psoriasis; therefore, the aim of treatment is to minimize the extent and severity of the disease to the point at which it no longer substantially disrupts the patient's quality of life. First-line therapy of psoriasis usually consists of topical agents, such as emollients, tar, dithranol, and vitamin D3 analogs. In cases of severe, extensive psoriasis, where topical therapy is either impractical or not sufficiently effective, systemic treatment may be warranted at the outset. In these circumstances, the therapeutic options include: (i) intensive inpatient or day center topical therapy; (ii) phototherapy; and/or (iii) systemic agents. There are now a number of systemic agents available for the treatment of severe psoriasis, but all have potential adverse effects. We review the current treatment options, which include the use of phototherapy and systemic agents, and provide recommendations on their use in clinical practice. Importantly, treatment should be tailored to each individual patient depending on concurrent medical problems (which might preclude certain agents), patient choice and acceptance of the risk of adverse effects. PMID- 16252929 TI - Pretibial myxedema: pathophysiology and treatment options. AB - Pretibial myxedema or localized myxedema or thyroid dermopathy is an autoimmune manifestation of Graves' disease. It also occasionally occurs in Hashimoto's thyroiditis. Lesions of thyroid dermopathy are usually asymptomatic and have only cosmetic importance. Advanced forms of dermopathy are associated with elephantiasis or thyroid acropachy. Almost all cases of thyroid dermopathy are associated with relatively severe ophthalmopathy. Usually ophthalmopathy appears first and dermopathy much later. All patients with localized myxedema have high serum concentrations of thyroid-stimulating hormone receptor antibodies, indicating the severity of the autoimmune condition. Occurrence of thyroid dermopathy in areas other than pretibial skin indicates a systemic process. Similar to Graves' ophthalmopathy, thyroid-stimulating hormone receptors in the connective tissue may be the antigen responsible for the immune process. Both humoral and cellular immune mechanisms are involved in the stimulation of fibroblasts and the production of large amounts of glycosaminoglycans. Localization in the pretibial area relates to mechanical factors and dependent position. Diagnosis of thyroid dermopathy is based on signs and typical pretibial skin lesions in association with a history of Graves' hyperthyroidism and ophthalmopathy. In some cases, skin biopsy is needed for confirmation. The lesions are usually mild and are overshadowed by more symptomatic ophthalmopathy. Most cases of thyroid dermopathy do not require any therapy. In mildly severe symptomatic cases and when there is cosmetic concern, topical corticosteroids applied under occlusive dressing are beneficial. In more severe cases, systemic immunomodulation may be necessary; however, conclusive evidence for long-term efficacy of these modalities is lacking. When significant edema and elephantiasis are present, local compressive therapy may have added benefit. In mild cases that do not require treatment, 50% of patients achieve complete remission after several years. Severe cases that receive topical corticosteroids or other therapies do not have a better outcome than untreated milder cases. Current treatment modalities for thyroid dermopathy and acropachy are at best palliative. Better and safer means of immunomodulation are needed. PMID- 16252930 TI - Topical halobetasol propionate in the treatment of plaque psoriasis: a review. AB - Halobetasol propionate (HP) 0.5% ointment and cream are class I topical corticosteroids. We review the efficacy and tolerability of HP for treatment of plaque psoriasis in the English language literature. The efficacy of HP ointment and cream is consistently superior to other super-potent topical corticosteroids. Local adverse events associated with topical HP are similar to those experienced with other super-potent corticosteroids. Combination therapy with calcipotriene (calcipotriol) and tazarotene appears to be superior to monotherapy with topical HP. PMID- 16252931 TI - Current management of herpes zoster: the European view. AB - The overall incidence of herpes zoster in Europe is approximately 3 per 1000 people per year and more than 10 per 1000 people per year in those aged >80 years. Post herpetic neuralgia (PHN) is a common debilitating complication of herpes zoster, particularly in patients aged >50 years, in persons with severe pain or rash at presentation, and in those with significant prodromal symptoms. Antiviral drugs can effectively control acute symptoms and, if used early enough in the course of the illness, can help prevent the development of PHN and other complications. However, despite this, many patients do not receive such treatment. The economic impact of zoster and PHN is largely underestimated in Europe. Furthermore, there is considerable variation throughout Europe in the management of herpes zoster. Use of antiviral therapy including the newer potent antiviral agents such as brivudin, which requires less frequent administration than acyclovir, is improving patient outcomes in some European countries. However, in many countries, patient awareness of herpes zoster and, as a result, overall antiviral use is low. Guidelines recommending the use of antiviral agents, particularly in patients at risk of developing PHN, are available but are not widely used. More needs to be done to educate the general public and increase awareness among primary healthcare providers of the benefits of timely and appropriate pharmacological therapy in patients with herpes zoster. PMID- 16252934 TI - Site-directed deep electronic tunneling through a molecular network. AB - Electronic tunneling in a complex molecular network of N(>2) donor/acceptor sites, connected by molecular bridges, is analyzed. The "deep" tunneling dynamics is formulated using a recursive perturbation expansion, yielding a McConnell-type reduced N-level model Hamiltonian. Applications to models of molecular junctions demonstrate that the donor-bridge contact parameters can be tuned in order to control the tunneling dynamics and particularly to direct the tunneling pathway to either one of the various acceptors. PMID- 16252932 TI - Treatment of pemphigus vulgaris: current and emerging options. AB - BACKGROUND: Pemphigus vulgaris is a rare, chronic, autoimmune mucocutaneous blistering disease. The disease can progress to involve the skin and multiple mucosae. Pemphigus vulgaris can be associated with a high morbidity and significant mortality rate. Treatment of the condition can be challenging. Conventional therapy primarily consists of systemic corticosteroids and immunosuppressant agents. In some patients with pemphigus vulgaris, these agents fail to provide an effective clinical response or have significant adverse effects. METHODS: We evaluated data on 792 patients with pemphigus vulgaris retrieved from PubMed, covering the period 1973-2004. Only patients reported in the English literature were included in this review. Recently, several new therapeutic agents and treatment modalities have been described for the treatment of patients with pemphigus vulgaris. Some therapeutic agents that were used in the past and abandoned have recently regained favor. This review focuses on the therapeutic uses of dapsone, methotrexate, mycophenolate mofetil, chlorambucil, dexamethasone-cyclophosphamide pulse therapy, immunoablative therapy with cyclophosphamide, plasmapheresis, and extracorporeal photochemotherapy. Newer agents, such as intravenous immunoglobulin (IVIg) therapy and rituximab (an anti CD20 chimeric monoclonal antibody), are also discussed. RESULTS AND CONCLUSIONS: Among the oral agents, dapsone may be considered a first-line agent. This is primarily because the risk of potentially fatal adverse effects with this drug is lower than that associated with other available chemotherapeutic agents. In patients who are refractory to oral agents, alternative treatments have been used to prevent further disease progression. Recently, the use of IVIg therapy, with a defined protocol, has been reported to be beneficial. This therapy is promising since it may allow for discontinuation of all other therapies and is safe. The adverse effects from IVIg therapy are minimal. Furthermore, compared with other therapies, it provides a better quality of life. PMID- 16252935 TI - Differences between solid superheating and liquid supercooling. AB - The thermodynamic and kinetic behaviors for solid superheating and liquid supercooling were critically examined and compared via molecular-dynamics simulations. It is shown that the large elastic energy associated with internal melting and solid-liquid interface disorder play important roles in superheating. The growth rate is anisotropic for supercooling, but isotropic for superheating. Supercooling can be well described by the classical nucleation theory, whereas superheating shows many exceptions. The underlying mechanisms for these differences are discussed. PMID- 16252936 TI - A density-functional model of the dispersion interaction. AB - We have recently introduced [J. Chem. Phys. 122, 154104 (2005)] a simple parameter-free model of the dispersion interaction based on the instantaneous in space, dipole moment of the exchange hole. The model generates remarkably accurate interatomic and intermolecular C6 dispersion coefficients, and geometries and binding energies of intermolecular complexes. The model involves, in its original form, occupied Hartree-Fock or Kohn-Sham orbitals. Here we present a density-functional reformulation depending only on total density, the gradient and Laplacian of the density, and the kinetic-energy density. This density-functional model performs as well as the explicitly orbital-dependent model, yet offers obvious computational advantages. PMID- 16252937 TI - A simplified relativistic time-dependent density-functional theory formalism for the calculations of excitation energies including spin-orbit coupling effect. AB - In the present work we have proposed an approximate time-dependent density functional theory (TDDFT) formalism to deal with the influence of spin-orbit coupling effect on the excitation energies for closed-shell systems. In this formalism scalar relativistic TDDFT calculations are first performed to determine the lowest single-group excited states and the spin-orbit coupling operator is applied to these single-group excited states to obtain the excitation energies with spin-orbit coupling effects included. The computational effort of the present method is much smaller than that of the two-component TDDFT formalism and this method can be applied to medium-size systems containing heavy elements. The compositions of the double-group excited states in terms of single-group singlet and triplet excited states are obtained automatically from the calculations. The calculated excitation energies based on the present formalism show that this formalism affords reasonable excitation energies for transitions not involving 5p and 6p orbitals. For transitions involving 5p orbitals, one can still obtain acceptable results for excitations with a small truncation error, while the formalism will fail for transitions involving 6p orbitals, especially 6p1/2 spinors. PMID- 16252938 TI - Correlated one-particle method: numerical results. AB - In a previous paper a correlated one-particle method was formulated, where the effective Hamiltonian was composed of the Fock operator and a correlation potential. The objective was to define a correlated one-particle theory that would give all properties that can be obtained from a one-particle theory. The Fock-space coupled-cluster method was used to construct the infinite-order correlation potential, which yields correct ionization potentials (IP's) and electron affinities (EA's) as the negative of the eigenvalues. The model, however, was largely independent of orbital choice. To exploit the degree of freedom of improving the orbitals, the Brillouin-Brueckner condition is imposed, which leads to an effective Brueckner Hamiltonian. To assess its numerical properties, the effective Brueckner Hamiltonian is approximated through second order in perturbation. Its eigenvalues are the negative of IP's and EA's correct through second order, and its eigenfunctions are second-order Brueckner orbitals. We also give expressions for its energy and density matrix. Different partitioning schemes of the Hamiltonian are used and the intruder state problem is discussed. The results for ionization potentials, electron affinities, dipole moments, energies, and potential curves are given for some sample molecules. PMID- 16252939 TI - Hardness and softness reactivity kernels within the spin-polarized density functional theory. AB - Generalized hardness and softness reactivity kernels are defined within a spin polarized density-functional theory (SP-DFT) conceptual framework. These quantities constitute the basis for the global, local (i.e., r-position dependent), and nonlocal (i.e., r and r'-position dependents) indices devoted to the treatment of both charge-transfer and spin-polarization processes in such a reactivity framework. The exact relationships between these descriptors within a SP-DFT framework are derived and the implications for chemical reactivity in such context are outlined. PMID- 16252940 TI - Convergence of replica exchange molecular dynamics. AB - Replica exchange molecular dynamics (REMD) method is one of the generalized ensemble algorithms which performs random walk in energy space and helps a system to escape from local energy traps. In this work, we studied the accuracy and efficiency of REMD by examining its ability to reproduce the results of multiple extended conventional molecular dynamics (MD) simulations and to enhance conformational sampling. Two sets of REMD simulations with different initial configurations, one from the fully extended and the other from fully helical conformations, were conducted on a fast-folding 21-amino-acid peptide with a continuum solvent model. Remarkably, the two REMD simulation sets started to converge even within 1.0 ns, despite their dramatically different starting conformations. In contrast, the conventional MD within the same time and with identical starting conformations did not show obvious signs of convergence. Excellent convergence between the REMD sets for T>300 K was observed after 14.0 ns REMD simulations as measured by the average helicity and free-energy profiles. We also conducted a set of 45 MD simulations at nine different temperatures with each trajectory simulated to 100.0 and 200.0 ns. An excellent agreement between the REMD and the extended MD simulation results was observed for T>300 K, showing that REMD can accurately reproduce long-time MD results with high efficiency. The autocorrelation times of the calculated helicity demonstrate that REMD can significantly enhance the sampling efficiency by 14.3+/-6.4, 35.1+/-0.2, and 71.5+/-20.4 times at, respectively, approximately 360, approximately 300, and approximately 275 K in comparison to the regular MD. Convergence was less satisfactory at low temperatures (T<300 K) and a slow oscillatory behavior suggests that longer simulation time was needed to reach equilibrium. Other technical issues, including choice of exchange frequency, were also examined. PMID- 16252941 TI - A density-functional study of the structural, electronic, magnetic, and vibrational properties of Ti8C12 metallocarbohedrynes. AB - Calculations are presented for the structural, electronic, and vibrational properties of the different Ti8C12 metallocarbohedrynes. (Please note that we adopt the name "metallocarbohedrynes" instead of "metallocarbohedrenes" to denote the acetylenic nature of C2 units in this class of clusters demonstrated by several contributions in literature.) The density-functional theory (DFT) calculations are performed with the all-electron projector augmented-wave method and generalized gradient approximation for the exchange-correlation functional. We study the seven low-energy isomers of the Ti8C12 metallocarbohedrynes using spin-polarized DFT, where we find a correlation between the number of rotated carbon dimers and the cohesive energy of the structure. The electronic density of states (eDOS) show that C3nu, D*3d, and D3d isomers are spin polarized. The partial eDOS shows that, depending on the dimer orientation, carbon atoms and a subgroup of the metal atoms form a covalent framework while other metal atoms are bonded to this framework more ionically. This picture is further supported by the charge density of the different structures, where we see that the Ti atoms with higher charge density show less contribution to the covalent bonding of the Ti-C framework. The vibrational spectra of the different structures are calculated using the frozen-vibration method. Also, we calculate the vibrational spectra of the C3nu and C2nu structures using molecular-dynamics simulations at two different temperatures. The results of the simulations demonstrate the local stability of the structures beyond the harmonic limit explored by the frozen vibration method. PMID- 16252942 TI - Fragmentation channels of large multicharged clusters. AB - We address unifying features of fragmentation channels driven by long-range Coulomb or pseudo-Coulomb forces in clusters, nuclei, droplets, and optical molasses. We studied the energetics, fragmentation patterns, and dynamics of multicharged (A+)n (n=55, 135, 321) clusters. In Morse clusters the variation of the range of the pair-potential induced changes in the cluster surface energy and in the fissibility parameter X=E(Coulomb)2E(surface). X was varied in the range of X=1-8 for short-range interactions and of X=0.1-1.0 for long-range interactions. Metastable cluster configurations were prepared by vertical ionization of the neutral clusters and by subsequent structural equilibration. The energetics of these metastable ionic clusters was described in terms of the liquid drop model, with the coefficients of the volume and surface energies depending linearly on the Morse band dissociation energy. Molecular-dynamics simulations established two distinct fragmentation patterns of multicharged clusters that involve cluster fission into a small number of large, multicharged clusters for X<1 and Coulomb explosion into a large number of individual ions and small ionic fragments for X>1. The Rayleigh instability limit X=1 separates between spatially anisotropic fission and spatially isotropic Coulomb explosion. Distinct features of the fragmentation energetics and dynamics were unveiled. For fission of n=55 clusters, large kinetic and internal energies of the large fragments are exhibited and the characteristic fragmentation time is approximately 700 fs, while for Coulomb explosion the major energy content of the small fragments involves kinetic energy and the characteristic fragmentation time of approximately 300 fs is shorter. The Rayleigh (X=1) limit, leading to isotropic Coulomb explosion, is transcended by a marked enhancement of the Coulomb energy, which is realized for extremely ionized clusters in ultraintense laser fields, or by a dramatic reduction of the surface energy as is the case for the expansion of optical molasses. PMID- 16252943 TI - Fully dimensional ab initio description of the structure and energetics of azabenzene-argon complexes. AB - The structure and energetics of van der Waals complexes of argon with azabenzenes: pyridine, pyrazine, pyrimidine, pyridazine, s-triazine, and s tetrazine are studied using the second-order Moller-Plesset perturbation theory combined with well-balanced basis sets. The full optimization of the cluster structures and computation of the inter- and intramolecular vibrational frequencies is performed by eliminating the basis set superposition error. The argon equilibrium coordinates are calculated with the accuracy comparable to that reached by standard methods of the structure determination from the spectral data. A simple rule to predict the position of argon with respect to the geometric center of the azabenzene ring is found. The calculated harmonic frequencies of the intermolecular vibrational modes are scaled by the factor of 0.85 to eliminate systematic errors coming from the neglect of anharmonic effects. The scaled frequencies agree with the experimental ones to about 1 cm( 1), except for pyrimidine-argon and tetrazine-argon for which empirical fundamental frequency estimates are problematic. A simple relation connecting the intermolecular bending frequencies and the monomer quadrupole tensor is found. The perturbation of the monomer properties caused by complexation is analyzed. The modification of the monomer structure by the interaction with argon and its influence on the binding energy appears to be negligible in all complexes studied. However, this interaction affects appreciably the intramolecular modes and causes their frequency shifts. As a consequence, the dissociation energy of the complexes increases by about 5 cm(-1). PMID- 16252944 TI - Improving the calculation of rovibrational spectra of five-atom molecules with three identical atoms by using a C3upsilonG6 symmetry-adapted grid: applied to CH3D and CHD3. AB - In this paper we report two improvements on the approach we have used to compute rovibrational levels of methane and apply the new ideas to calculate rovibrational levels of two methane isotopomers CH3D and CHD3. Both improvements make the bend calculation better. The first improvement is a G6-invariant (or C3upsilon-invariant) grid which is designed such that each point on the grid is mapped to another point on the grid by any of the G6 operations. The second improvement is the use of fast Fourier transform (FFT) to compute the bend potential matrix-vector products. The FFT matrix-vector product is about three and ten times faster than the previous sequential summation method for the J=0 and J>0 cases, respectively. The calculated J=1 rovibrational levels of CH3D and CHD3 on the Schwenke and Partridge [Spectrochim. Acta, Part A 57, 887 (2001)] ab initio potential are in good agreement (within 6 cm(-1) for the levels up to 3000 cm(-1)) with the experimental data. The agreement is even better (within 0.1 cm( 1) for the levels up to 6000 cm(-1)) if the associated J=0 energies are subtracted. PMID- 16252945 TI - Photoionization of small krypton clusters in the Kr 3d regime: evidence for site specific photoemission. AB - Kr 3d ionization energies of small, variable size krypton clusters are investigated by photoelectron spectroscopy, where the size regime of clusters with an average size N< or =30 is studied. Characteristic shifts in Kr 3d ionization energies to lower binding energies are found compared to the bare atom. These are also different from those of large krypton clusters. Moreover, we find evidence for photoionization of the krypton dimer. Its 3d ionization energy is barely shifted relative to the atomic value. Results from model calculations considering different isomers and cluster sizes as well as defect sites give evidence that the experimental results can be related to photoionization from different surface sites in variable size krypton clusters. This can be related to site-specific photoemission in small Kr clusters. The results are compared to size effects in Kr 3d near-edge features of variable size Kr clusters as well as recent results on Kr 3d photoionization of large Kr clusters. PMID- 16252946 TI - Concerted and sequential Coulomb explosion processes of N2O in intense laser fields by coincidence momentum imaging. AB - The Coulomb explosion dynamics of N2O in intense laser fields (800 nm, 60 fs, approximately 0.16 PWcm2) is studied by the coincidence momentum imaging method. From the momentum correlation maps obtained for the three-body fragmentation pathway, N2O3+-->N++N++O+, the ultrafast structural deformation dynamics of N2O prior to the Coulomb explosion is extracted. It is revealed that the internuclear N-N and N-O distances stretch simultaneously as the bond angle less than approximately N-N-O decreases. In addition, two curved thin distributions are identified in the momentum correlation maps, and are interpreted well as those originating from the sequential dissociation pathway, N2O3+-->N++NO2+-->N++N++O+. PMID- 16252947 TI - Nanosecond time-resolved IR emission from molecules excited in a supersonic jet: intramolecular dynamics of NO2 near dissociation. AB - IR emission from NO2 cooled in a supersonic jet and excited to a single, B 2B1 state rovibronic level at 22 994.92 cm(-1) above the ground-state zero point was detected with 10(-8)-s time resolution. The IR emission together with the laser induced fluorescence decay measurement allows the deduction of the relaxation dynamics near the dissociation of NO2. Following the excitation this single rovibronic B 2B1 level decays on 1.0-s time scale primarily through electronic radiation. Collisions induce internal conversion with a rate constant of 3.0 x 10(7) Torr(-1) s(-1) to the mixed AX states. Collisions further induce internal conversion of the AX mixed states into highly vibrationally excited levels in the X states with a rate constant at least one order of magnitude slower. This mechanism results in the observation of a double-exponential decay in the laser induced fluorescence and a rise in the IR emission intensity corresponding to the fast decay in the fluorescence intensity. The IR emission rate of the highly vibrationally excited X-state levels is estimated to be about one order of magnitude larger than the isoenergetic AX mixed states and much larger than the B 2B1 level, both with much less vibrational excitation. PMID- 16252948 TI - Continuous-wave cavity ringdown spectroscopy of the 8nu polyad of water in the 25,195-25,340 cm(-1) range. AB - State-of-the-art experiments and calculations are used to record and assign the data obtained in the weakly absorbing blue energy region of the H2O spectrum. Continuous-wave cavity ringdown absorption spectroscopy with Doppler resolution is used to probe the range from 25,195 to 25,470 cm(-1) with an absorption sensitivity of approximately 1 parts per 10(9) (ppb)/cm. 62 lines of the polyad nu(OH)=8 are reported, of which 43 are assigned using variational nuclear calculations. The study includes absorption line intensities (in the range of 10( 28)-10(-26) cmmolecule) for all lines and self-broadening pressure coefficient for a few lines. The newly obtained energy levels are also reported. PMID- 16252949 TI - Competitive charge- and energy-transfer processes following core ionization in the Na- CO cluster. AB - Anion-molecule clusters constitute a very suitable class of systems for studying intermolecular (interatomic) charge-transfer (CT) processes following core ionization. A weakly bound electron of the anion in these clusters can be easily transferred to the core-ionized molecule. The screening effect of this electron may have a dramatic impact on core-level spectra and even account for a breakdown of the quasiparticle picture of core ionization. This is demonstrated here by calculating the O1s(-1) and C1s(-1) core ionization spectra of the Na- CO cluster using an ab initio fourth-order Green's-function method. Interestingly, along with the CT processes in this cluster there exist also very efficient energy transfer (ET) processes favored by the low excitation energies of Na-. These ET processes constitute an appreciable part of the electronic excitations following core ionization of Na- CO and exert thereby a strong influence on the spectra studied. The spectral features attributed to the ET processes are as pronounced as those attributed to the CT processes. Major differences in the behavior of CT and ET satellites as a function of the anion-molecule separation are found and explained. We compare also the O1s(-1) core ionization spectra of the Na- CO and Na- H2O clusters. Along with a certain similarity, these spectra exhibit substantial differences which are essentially attributed to the distinct cluster geometries. PMID- 16252950 TI - Rotational Raman spectroscopy of ethylene using a femtosecond time-resolved pump probe technique. AB - Femtosecond Raman-induced polarization spectroscopy (RIPS) was conducted at low pressure (250 mb at 295 K and 400 mb at 373 K) in ethylene. The temporal signal, resulting from the beating between pure rotational coherences, was measured with a heterodyne detection. The temporal traces were converted to the frequency domain using a Fourier transformation and then analyzed thanks to the D2hTDS software (http://www.u-bourgogne.fr/LPUB/shTDS.html) dedicated to X2Y4 molecules with D2h symmetry. The effective Hamiltonian was expanded up to order 2, allowing the determination of five parameters with an rms of 0.017 cm(-1). Special care was taken in the precise modeling of intensities, taking into account all instrumental effects. Relative intensities were fitted (with an rms of 7.2%) and two polarizability operators were determined. PMID- 16252951 TI - Studies of collisional selection rules in thioformaldehyde (H2CS) by microwave optical double resonance. AB - Individual rotational levels in the A 1A2,v4=1 state of thioformaldehyde (H2CS) are excited by a cw laser and microwave transitions in the region of 8-12 GHz are measured. Some of the microwave frequencies are found to be characteristic of rotational levels other than the level being pumped. Since the microwave frequencies are characteristic of individual rotational levels in the excited state and the excited-state lifetime is approximately 170 micros, information is obtained concerning rotational selection rules during collisions. It is found that J can change by several units and Ka by 0, +/-2, +/-4, and +/-6. The latter result confirms that o-H2CS is not converted to p-H2CS by collisions. Observation of Ka doublets indicates that there are no appreciable differences between the two components. PMID- 16252952 TI - Quasiclassical dynamics simulation of the collision-induced dissociation of Cr(CO)6 + with Xe. AB - Quasiclassical trajectory calculations are employed to investigate the dynamics of collision-induced dissociation (CID) of Cr(CO)6 + with Xe atoms at collision energies ranging from 1.3 to 5.0 eV. The trajectory simulations show that direct elimination of CO ligands, during the collision, becomes increasingly important as the collision energy increases. In a significant number of cases, this shattering mechanism is accompanied with a concomitant formation of a transient Xe-Cr(CO)x +(x<6) complex. The calculated results are in very good agreement with the experimental results presented previously [F. Muntean and P. B. Armentrout, J. Chem. Phys. 115, 1213 (2001)]. In particular, the computed cross sections and scattering maps for the product ions Cr(CO)x +(x=3-5) compare very favorably with the reported experimental data. However, in contrast with the conclusions of the previous study, the present calculations suggest that CID dynamics for this system exhibits a significant impulsive character rather than proceeding via a complex surviving more than a rotational period. PMID- 16252954 TI - Continuation calculations of boron- (aluminum-, titanium-, and nickel-) doped La13 clusters. AB - In this work, we have calculated boron-, aluminum-, titanium-, and nickel-doped La13 clusters by DMOL method based on the density-functional theory. Two doping modes are employed: surface and center doping. The boron, aluminum, and nickel atoms prefer to occupy the surface sites while the titanium atom prefers to occupy the center site. The doped La13 clusters with these four kinds of atoms have lower binding energy than pure La13 clusters. The icosahedral isomers are of lower binding energy than cubotahedral and decahedral isomers for La12B(-1), La12Al(-1), and La12Ni, while doping makes the cubotahedral La12Ti stable with a binding energy a little lower than icosahedral La12Ti. There are electronic shell effects in icosahedral La12B(-1) and La12Al(-1). The icosahedral La12B(-1) is promising to be formed during the doped process experimentally. Furthermore, we have also discussed the distorted structures of center doping by bond lengths, density of states, and charge transfers. PMID- 16252953 TI - An analysis of the electrostatic interaction between nucleic acid bases. AB - Results from several commonly used approximate methods of evaluating electrostatic interactions have been compared to the rigorous, nonexpanded electrostatic energies at both uncorrelated and correlated levels of theory. We examined a number of energy profiles for both hydrogen bonded and stacked configurations of the nucleic acid base pairs. We found that the penetration effects play an extremely important role and the expanded electrostatic energies are significantly underestimated with respect to the ab initio values. Apart from the inability to reproduce the magnitudes of the ab initio electrostatic energy, there are other problems with the available approximate electrostatic models. For example, the distributed multipole analysis, one of the most advanced methods, is extremely sensitive to the basis set and level of theory used to evaluate the multipole moments. Detailed ab initio results are provided that other researchers could use to test their approximate models. PMID- 16252955 TI - Molecular-dynamics study of the density scaling of inert gas condensation. AB - The initial stages of vapor condensation of Ge in the presence of a cold Ar atmosphere were studied by molecular-dynamics simulations. The state variables of interest included the densities of condensing vapor and gas, the density of clusters, and the average cluster size, while the temperatures of the vapor and the clusters were separately monitored with time. Three condensation processes were explicitly identified: nucleation, monomeric growth, and cluster aggregation. Our principal finding is that both the average cluster size and the number of clusters scale with the linear dimension of the computation cell, L, and Ln, with the scaling parameter n approximately 4, corresponding to a reaction order of nu approximately 2.33. This small value of n is explained by an unexpected nucleation path involving the formation of Ge dimers via two-body collisions. PMID- 16252956 TI - Transient cavities and the excess chemical potentials of hard-spheroid solutes in dipolar hard-sphere solvents. AB - Monte Carlo computer simulations are used to study transient cavities and the solvation of hard-spheroid solutes in dipolar hard-sphere solvents. The probability distribution of spheroidal cavities in the solvent is shown to be well described by a Gaussian function, and the variations of fit parameters with cavity elongation and solvent properties are analyzed. The excess chemical potentials of hard-spheroid solutes with aspect ratios x in the range of 15< or =x< or =5, and with volumes between 1 and 20 times that of a solvent molecule, are presented. It is shown that for a given molecular volume and solvent dipole moment (or temperature) a spherical solute has the lowest excess chemical potential and hence the highest solubility, while a prolate solute with aspect ratio x should be more soluble than an oblate solute with aspect ratio 1x. For a given solute molecule, the excess chemical potential increases with increasing temperature; this same trend can be observed in hydrophobic solvation. A scaled particle theory based on the solvent equation of state and a fitted solute solvent interfacial tension shows excellent agreement with the simulation results over the whole range of solute elongations and volumes considered. An information theoretic model based on the solvent density and radial distribution function is less successful, being accurate only for small solute volumes and low solvent densities. PMID- 16252957 TI - On the dielectric susceptibility spectra of supercooled o-terphenyl. AB - Supercooled o-terphenyl has been the subject of many investigations including dielectric relaxation spectroscopy. Due to the low dielectric strength and the tendency to crystallize at elevated temperatures, a detailed shape analysis of the loss profile from the glass transition temperature Tg to approximately 1.2 Tg is not available for the neat glass former. Assessing the origin of the different temperature dependencies of translational and rotational motions in supercooled liquids and its possible connection to heterogeneity requires this knowledge regarding the possible changes in the relaxation-time distribution across the 100 s-100 ns relaxation-time range. This note provides this information for o terphenyl on the basis of a master curve representation: time-temperature superposition applies with a constant stretching exponent of beta=0.5 in the range of interest. PMID- 16252959 TI - Quantum diffusion in liquid water from ring polymer molecular dynamics. AB - We have used the ring polymer molecular-dynamics method to study the translational and orientational motions in an extended simple point charge model of liquid water under ambient conditions. We find, in agreement with previous studies, that quantum-mechanical effects increase the self-diffusion coefficient D and decrease the relaxation times around the principal axes of the water molecule by a factor of around 1.5. These results are consistent with a simple Stokes-Einstein picture of the molecular motion and suggest that the main effect of the quantum fluctuations is to decrease the viscosity of the liquid by about a third. We then go on to consider the system-size scaling of the calculated self diffusion coefficient and show that an appropriate extrapolation to the limit of infinite system size increases D by a further factor of around 1.3 over the value obtained from a simulation of a system containing 216 water molecules. These findings are discussed in light of the widespread use of classical molecular dynamics simulations of this sort of size to model the dynamics of aqueous systems. PMID- 16252958 TI - Spectroscopic characterization of microscopic hydrogen-bonding disparities in supercritical water. AB - The local hydrogen-bonding environment in supercritical water (380 degrees C, 300 bars, density 0.54 gcm3) was studied by x-ray Raman scattering at the oxygen K edge. The spectra are compared to those of the gas phase, liquid surface, bulk liquid, and bulk ice, as well as to calculated spectra. The experimental model systems are used to assign spectral features and to quantify specific local hydrogen-bonding situations in supercritical water. The first coordination shell of the molecules is characterized in more detail with the aid of the calculations. Our analysis suggests that approximately 65% of the molecules in supercritical water are hydrogen bonded in configurations that are distinctly different from those in liquid water and ice. In contrast to liquid water the bonded molecules in supercritical water have four intact hydrogen bonds and in contrast to ice large variations of bond angles and distances are observed. The remaining approximately 35% of the molecules exhibit two free O-H bonds and are thus either not involved in hydrogen bonding at all or have one or two hydrogen bonds on the oxygen side. We determine an average O-O distance of 3.1+/-0.1 A in supercritical water for the H bonded molecules at the conditions studied here. This and the corresponding hydrogen bond lengths are shown to agree with neutron- and x-ray-diffraction data at similar conditions. Our results on the local hydrogen-bonding environment with mainly two disparate hydrogen-bonding configurations are consistent with an extended structural model of supercritical water as a heterogeneous system with small patches of bonded molecules in various tetrahedral configurations and surrounding nonbonded gas-phase-like molecules. PMID- 16252960 TI - Primitive model for cation hydrolysis: a molecular-dynamics study. AB - A model of primitive cation MZ+ in water is introduced in order to clarify the influence of ion charge on the hydration structure and dynamic properties of highly charged cations in aqueous solutions. A flexible nonconstrained model for water molecules is used. The considered model in the case of monovalent cation M+ reduces to the realistic model for the hydration structure of Na+. It is shown that for divalent ion M2+ the strong cation-water electrostatic interaction leads to the formation of stable structures constituted by six water molecules octahedrally arranged around the cation. The cation-oxygen attraction and cation hydrogen repulsion modify the octahedral hydration configuration of the model cations M3+ and M4+ and additional water molecules can join the hydration shell. The increase of cation charge results in the increase of O-H bond length of water molecules in the cation hydration shell. Further increase of ion-water electrostatic interaction causes the loss of some protons from the hydration shell of cations M4+, M5+, and M6+ that is interpreted as a cation hydrolysis effect. For a correct description of this phenomenon the considered model is improved by modeling the effects of the charge redistribution between hydrolysis products, which essentially modified and stabilized the hydrated-hydrolyzed structure of cation. The influence of cation charge on dynamical properties of cation MZ+ and oxygens in its hydration shell was investigated and analyzed. PMID- 16252961 TI - Application of polyamorphism in water to spontaneous crystallization of emulsified LiCl-H2O solution. AB - Aqueous solutions are widely explained by the hydration or the bound waterfree water notion. Amorphous polymorphism (polyamorphism) in pure water, which is presently under vigorous discussion, may provide a different view over the solutions. Here, I changed pressure, P, temperature, T, and concentration, C, of emulsified LiCl-H2O solutions and studied their freezing by detecting its heat evolution. It was experimentally indicated that the homogeneous nucleation of low density crystalline ice I (phase Ih or Ic), in pure water and in solutions, connects to the polyamorphic transition of high-density amorphous ice (HDA) to low-density amorphous ice (LDA). Thus, the polyamorphism of water relates to the phase behavior of aqueous solution. In accordance with the recent simulation result, the nucleation was thought to occur in two stages: the appearance of the LDA-like state and the crystallization. Usefulness of the polyamorphic point of view about the solutions was seen. PMID- 16252962 TI - Molecular-dynamics simulations of dimethylsulfoxide-methanol mixtures. AB - We present molecular-dynamics (MD) computer simulation results for the local structures, hydrogen (H)-bond distribution, and dynamical properties of methanol (MeOH) and dimethylsulfoxide (DMSO) binary mixtures at ambient conditions over the entire composition range. The simulated heat of mixing and site-site pair distribution functions suggest that the intermolecular structures of the pure liquids are not markedly altered upon mixing. Nevertheless, H-bonding statistics show that aggregates of the type 1DMSO:1MeOH are formed and represent the predominant form of molecular association in these mixtures. Only a small fraction (10%) of DMSO molecules in MeOH-rich mixtures (85% in mole) forms H bonding trimers of type 1DMSO:2MeOH. No evidence of other types of interspecies association is found. The self-diffusion coefficient for DMSO (MeOH) increases (decreases) upon mixing. The characteristic reorientation time tau1 of both species increases in the mixture, but the composition dependence is weak. The frequency spectrum of MeOH reorientational time-correlation function shows significant redshifts of the principal librational band as DMSO is added to the system, whereas the librational band of DMSO shows small alterations upon mixing. Our results are discussed in the light of previous simulation analyses for a similar system, DMSO-water mixtures, and compared with available experimental results. PMID- 16252963 TI - Pore nucleation in mechanically stretched bilayer membranes. AB - We report a computer-simulation study of the free-energy barrier for the nucleation of pores in the bilayer membrane under constant stretching lateral pressure. We find that incipient pores are hydrophobic but as the lateral size of the pore nucleus becomes comparable with the molecular length, the pore becomes hydrophilic. In agreement with previous investigations, we find that the dynamical process of growth and closure of hydrophilic pores is controlled by the competition between the surface tension of the membrane and the line tension associated with the rim of the pore. We estimate the line tension of a hydrophilic pore from the shape of the computed free-energy barriers. The line tension thus computed is in a good agreement with available experimental data. We also estimate the line tension of hydrophobic pores at both macroscopic and microscopic levels. The comparison of line tensions at these two different levels indicates that the "microscopic" line tension should be carefully distinguished from the "macroscopic" effective line tension used in the theoretical analysis of pore nucleation. The overall shape of the free-energy barrier for pore nucleation shows no indication for the existence of a metastable intermediate during pore nucleation. PMID- 16252964 TI - T dependence of vibrational dynamics of water in ion-exchanged zeolites A: a detailed Fourier transform infrared attenuated total reflection study. AB - In order to explore the influence of cation substitution on the vibrational dynamics of water molecules in zeolites, the evolution of structural properties of the O-H stretching band of water in fully hydrated Na-A and Mg-exchanged A zeolites has been studied, for different percentages of induced ion exchange, by Fourier transform infrared attenuated total reflection spectroscopy as a function of temperature. The differences revealed in the O-H stretching band shapes have been accounted by fitting the spectra as a sum of four components, corresponding to water molecules exhibiting different types of hydrogen bonding. The dependencies of the relative intensities, peak wave numbers, and bandwidths of the resolved components on temperature and Mg2+ content have been discussed. Evidence of the "structure-maker" role played by a zeolitic surface on physisorbed water, systematically enhanced by increasing the percentage of induced ion exchange, is given in the whole explored temperature range. PMID- 16252965 TI - Atomic force microscopy measurements of topography and friction on dotriacontane films adsorbed on a SiO2 surface. AB - We report comprehensive atomic force microscopy (AFM) measurements at room temperature of the nanoscale topography and lateral friction on the surface of thin solid films of an intermediate-length normal alkane, dotriacontane (n C32H66), adsorbed onto a SiO2 surface. Our topographic and frictional images, recorded simultaneously in the contact mode, reveal a multilayer structure in which one to two layers of molecules adsorb adjacent to the SiO2 surface oriented with their long axis parallel to the interface followed by partial layers of molecules oriented perpendicular to the surface. The thicknesses of the parallel and perpendicular layers that we measured with the AFM agree with those inferred from previous x-ray specular reflectivity measurements on similarly prepared samples. We also observe bulk dotriacontane particles and, in contrast with our previous measurements, are able to determine their location. Above a minimum size, the bulk particles are separated from islands of perpendicularly oriented molecules by regions of exposed parallel layers that most likely extend underneath the particles. We find that the lateral friction is sensitive to the molecular orientation in the underlying crystalline film and can be used effectively with topographic measurements to resolve uncertainties in the film structure. We measure the same lateral friction on top of the bulk particles as on the perpendicular layers, a value that is about 2.5 times smaller than on a parallel layer. Scans on top of parallel layers indicate a constant height but reveal domains having different sublevels of friction. We explain this by the domains having different azimuthal orientations of the molecules. PMID- 16252966 TI - Monte Carlo calculations for the intrinsic viscosity of several dendrimer molecules. AB - We have performed Monte Carlo simulations to reproduce the intrinsic viscosity corresponding to different generation of several types of dendrite molecules: polyamidoamine dendrimers with an ethylendiamine core, polypropylene-imine with a diaminobutane core, and monodendrons and tridendrons of polybenzylether. With this end, we have employed coarse-grained idealizations of the molecules constituted by only two beads in each repeat unit (one in a branching or end unit and one intermediate along the repeat unit) and a simple hard-sphere potential between non-neighboring beads. Our goal is to investigate if this simple model is able to provide a reasonable description of some differences between these systems that have been observed experimentally, in particular, the location of the maximum in the intrinsic viscosity as a function of the generation number. Experimental radii of gyration in a given solvent are reproduced by a fit of the hard-sphere potential diameter. Subsequently, intrinsic viscosities are calculated by the variational approach of Fixman, which yields an accurate lower bound value with an additional hydrodynamic interaction parameter (the friction radius of the beads). The results show a pronounced variation of the maximum location with the value of the friction radius and the structural details that cannot be mimicked with simpler models. The initial conformations for the Monte Carlo procedure are taken from atomistic configurations thermalized by means of a molecular dynamics. PMID- 16252967 TI - Frequency-dependent dielectric permittivity of salt-free charged lamellar systems. AB - We present a new model to analyze dielectric spectroscopy measurements on charged lamellar systems, with the following improvements with respect to the hitherto available models: (i) it does not rely on the hypothesis of local electro neutrality, and allows to treat the salt-free case; (ii) the chemical exchange governing the partition between free and bound ions is properly taken into account; (iii) a fully analytical solution is provided. The variation of the frequency-dependent dielectric permittivity with both thermodynamic and kinetic characteristics of the free-bound ion equilibrium is presented. In particular, the relative weights of both relaxation modes (exchange and transport), and their characteristic frequencies are discussed. This study opens the way to the analysis of systems for which the usual models are irrelevant, such as salt-free clay gels or membranes. PMID- 16252968 TI - Thermodynamics of Go-type models for protein folding. AB - Go-type potentials, based on the inter-residue contacts present in the native structure of a protein, are frequently used to predict dynamic and structural features of the folding pathways through computer simulations. However, the mathematical form used to define the model interactions includes several arbitrary choices, whose consequences are not usually analyzed. In this work, we use a simple off-lattice protein model and a parallel tempering Monte Carlo simulation technique to carry out such analysis, centered in the thermodynamic characteristics of the folding transition. We show how the definition of a native contact has a deep impact on the presence of simple or complex transitions, with or without thermodynamic intermediates. In addition, we have checked that the width of the attractive wells has a profound effect on the free-energy barrier between the folded and unfolded states, mainly through its influence on the entropy of the denatured state. PMID- 16252969 TI - The polymerization of actin: structural changes from small-angle neutron scattering. AB - We present a new analysis of small-angle neutron-scattering data from rabbit muscle actin in the course of the polymerization from G-actin to F-actin as a function of temperature. The data, from Ivkov et al. [J. Chem. Phys. 108, 5599 (1998)], were taken in D2O buffer with Ca2+ as the divalent cation on the G-actin in the presence of ATP and with KCl as the initiating salt. The new analysis of the data using modeling and the method of generalized indirect fourier transform (O. Glatter, GIFT, University of Graz, Austria, http://physchem.kfunigraz.ac.at/sm/) provide shapes and dimensions of the G-actin monomer and of the growing actin oligomer in solution as a function of temperature and salt concentration. This analysis indicates that the G-actin monomer, under the conditions given above, is a sphere 50-54 A in diameter as opposed to the oblate ellipsoid seen by x-ray crystallography. The F-actin dimensions are consistent with x-ray crystal structure determinations. PMID- 16252970 TI - Gel mechanics: a comparison of the theories of Biot and Tanaka, Hocker, and Benedek. AB - This article compares the Biot [J. Appl. Phys. 12, 155 (1941)] and Tanaka, Hocker, and Benedek (THB) [J. Chem. Phys. 59, 5151 (1973)] theories of gel deformation. Biot's theory treats the gel as a continuum with the pore pressure as a state variable whereas the THB theory treats the gel as a mixture of solid and liquid phases. We revisit the problem of gel deswelling and use this example to show that there can be substantial differences between the two theories. The THB theory is not a complete mixture theory since the displacements of the liquid are assumed to be negligibly small in comparison with the displacements of the network. We propose a simple extension of the THB model, which takes into account the momentum transfer of the liquid phase. We show that with this simple addition and some very reasonable assumptions, the extended THB theory is identical to the Biot theory. PMID- 16252971 TI - Topology-dependent protein folding rates analyzed by a stereochemical model. AB - It is an experimental fact that gross topological parameters of the native structure of small proteins presenting two-state kinetics, as relative contact order chi, correlate with the logarithm of their respective folding rate constant kappa(f). However, reported results show specific cases for which the (chi,log kappa(f)) dependence does not follow the overall trend of the entire collection of experimental data. Therefore, an interesting point to be clarified is to what extent the native topology alone can explain these exceptional data. In this work, the structural determinants of the folding kinetics are investigated by means of a 27-mer lattice model, in that each native is represented by a compact self-avoiding (CSA) configuration. The hydrophobic effect and steric constraints are taken as basic ingredients of the folding mechanism, and each CSA configuration is characterized according to its composition of specific patterns (resembling basic structural elements such as loops, sheets, and helices). Our results suggest that (i) folding rate constants are largely influenced by topological details of the native structure, as configurational pattern types and their combinations, and (ii) global parameters, as the relative contact order, may not be effective to detect them. Distinct pattern types and their combinations are determinants of what we call here the "content of secondary type" structure (sigma) of the native: high sigma implies a large kappa(f). The largest part of all CSA configurations presents a mix of distinct structural patterns, which determine the chixlog kappa(f) linear dependence: Those structures not presenting a proper chi-dependent balance of patterns have their folding kinetics affected with respect to the pretense linear correlation between chi and log kappa(f). The basic physical mechanism relating sigma and kappa(f) involves the concept of cooperativity: If the native is composed of patterns producing a spatial order rich in effective short-range contacts, a properly designed sequence undertakes a fast folding process. On the other hand, the presence of some structural patterns, such as long loops, may reduce substantially the folding performance. This fact is illustrated through natives having a very similar topology but presenting a distinct folding rate kappa(f), and by analyzing structures having the same chi but different sigma. PMID- 16252972 TI - Population dynamics simulations of functional model proteins. AB - In order to probe the fundamental principles that govern protein evolution, we use a minimalist model of proteins to provide a mapping from genotype to phenotype. The model is based on physically realistic forces of protein folding and includes an explicit definition of protein function. Thus, we can find the fitness of a sequence from its ability to fold to a stable structure and perform a function. We study the fitness landscapes of these functional model proteins, that is, the set of all sequences mapped on to their corresponding fitnesses and connected to their one mutant neighbors. Through population dynamics simulations we directly study the influence of the nature of the fitness landscape on evolution. Populations are observed to move to a steady state, the distribution of which can often be predicted prior to the population dynamics simulations from the nature of the fitness landscape and a quantity analogous to a partition function. In this paper, we develop a scheme for predicting the steady-state population on a fitness landscape, based on the nature of the fitness landscape, thereby obviating the need for explicit population dynamics simulations and providing some insight into the impact on molecular evolution of the nature of fitness landscapes. Poor predictions are indicative of fitness landscapes that consist of a series of weakly connected sublandscapes. PMID- 16252973 TI - Statistical theory for protein ensembles with designed energy landscapes. AB - Combinatorial protein libraries provide a promising route to investigate the determinants and features of protein folding and to identify novel folding amino acid sequences. A library of sequences based on a pool of different monomer types are screened for folding molecules, consistent with a particular foldability criterion. The number of sequences grows exponentially with the length of the polymer, making both experimental and computational tabulations of sequences infeasible. Herein a statistical theory is extended to specify the properties of sequences having particular values of global energetic quantities that specify their energy landscape. The theory yields the site-specific monomer probabilities. A foldability criterion is derived that characterizes the properties of sequences by quantifying the energetic separation of the target state from low-energy states in the unfolded ensemble and the fluctuations of the energies in the unfolded state ensemble. For a simple lattice model of proteins, excellent agreement is observed between the theory and the results of exact enumeration. The theory may be used to provide a quantitative framework for the design and interpretation of combinatorial experiments. PMID- 16252974 TI - Monte Carlo simulation of Li+ motion in polyethylene based on polarization energy calculations and informed by data compression analysis. AB - We present an n-fold way kinetic Monte Carlo simulation of the hopping motion of Li+ ions in polyethylene on a grid of mesh 0.36 A superimposed on the voids of the rigid polymer. The structure of the polymer is derived from a higher-order simulation, and the energy of the ion at each site is derived by the self consistent polarization field method. The ion motion evolves in time from free flight through anomalous diffusion to normal diffusion, with the average energy tending to decrease with increasing temperature through thermal annealing. We compare the results with those of hopping models with probabilistic energy distributions of increasing complexity by analyzing the mean-square displacement and the average energy of an ensemble of ions. The Gumbel distribution describes the ion energy statistics in this system better than the usual Gaussian distribution does; including energy correlation greatly affects the ion dynamics. The analysis uses the standard data compression program GZIP, which proves to be a powerful tool for data analysis by giving a measure of recurrences in the ion path. PMID- 16252975 TI - Spread spectrum communication system with chaotic frequency modulation. AB - A new spread spectrum communication system utilizing chaotic frequency modulation of sinusoidal signals is discussed. A single phase lock loop (PLL) system in the receiver is used both to synchronize the local chaotic oscillator and to recover the information signal. We study the dynamics of the synchronization process, stability of the PLL system, and evaluate the bit-error-rate performance of this chaos-based communication system. PMID- 16252976 TI - Constructing dynamical systems with specified symbolic dynamics. AB - In this paper we demonstrate how to construct signals (time series) of continuous time dynamical systems that exhibit a given symbolic dynamics. This is achieved without construction of the ordinary differential equations that generate the flow. This construction is of theoretical interest and is useful as a source of dynamical data that can be used to test various data analysis algorithms. PMID- 16252977 TI - Bistability in a complementary metal oxide semiconductor inverter circuit. AB - Radiofrequency signals can disrupt the operation of low frequency circuits. A digital inverter circuit would seem to be immune to such disruption, because its output state usually jumps abruptly between 0 and 5 V. Nevertheless, when driven with a high frequency signal, the inverter can have two coexisting stable states (which are not at 0 and 5 V). Slow switching between these states (by changing the rf signal) will produce a low frequency signal. I demonstrate the bistability in a circuit experiment and in a simple model of the circuit. PMID- 16252978 TI - Multiple time scale chaos in a Schmitt trigger circuit. AB - It is known that stray radio frequency signals can produce nonlinear effects that disrupt the operation of circuits, but the mechanisms by which this disruption occurs are not well known. In this paper, an emitter coupled Schmitt trigger circuit is driven with a high-frequency signal to look for disruptive effects. As the circuit makes a transition between mode locked states (period 2 and period 3, for example), there is a region of chaos in which the largest peak in the power spectrum is in between the mode-locked frequencies, and is not related to the driving frequency by an integer multiple. This chaos resembles the chaos seen during a period adding sequence, except that it contains frequencies ranging over many orders of magnitude, from the driving frequencies on the order of megahertz, down to a few hertz. It is found that only a one-transistor circuit is necessary to produce this extremely broadband chaos, and true quasiperiodicity is not seen in this circuit. The single-transistor circuit is then simulated to confirm the frequency conversion effects. PMID- 16252979 TI - Stickiness in mushroom billiards. AB - We investigate the dynamical properties of chaotic trajectories in mushroom billiards. These billiards present a well-defined simple border between a single regular region and a single chaotic component. We find that the stickiness of chaotic trajectories near the border of the regular region occurs through an infinite number of marginally unstable periodic orbits. These orbits have zero measure, thus not affecting the ergodicity of the chaotic region. Notwithstanding, they govern the main dynamical properties of the system. In particular, we show that the marginally unstable periodic orbits explain the periodicity and the power-law behavior with exponent gamma=2 observed in the distribution of recurrence times. PMID- 16252980 TI - Correlation dimension and integral do not predict epileptic seizures. AB - Reports in the literature have indicated potential value of the correlation integral and dimension for prediction of epileptic seizures up to several minutes before electrographic onset. We apply these measures to over 2000 total hours of continuous electrocortiogram, taken from 20 patients with epilepsy, examine their sensitivity to quantifiable properties such as the signal amplitude and autocorrelation, and investigate the influence of embedding and filtering strategies on their performance. The results are compared against those obtained from surrogate time series. Our conclusion is that neither the correlation dimension nor the correlation integral has predictive power for seizures. PMID- 16252981 TI - Covariant Hamiltonian for the electromagnetic two-body problem. AB - We give a Hamiltonian formalism for the delay equations of motion of the electromagnetic two-body problem with arbitrary masses and with either repulsive or attractive interaction. This dynamical system based on action-at-a-distance electrodynamics appeared 100 years ago and it was popularized in the 1940s by the Wheeler and Feynman program to quantize it as a means to overcome the divergencies of perturbative QED. Our finite-dimensional implicit Hamiltonian is closed and involves no series expansions. As an application, the Hamiltonian formalism is used to construct a semiclassical canonical quantization based on the numerical trajectories of the attractive problem. PMID- 16252982 TI - A new route to chaos: sequences of topological torus bifurcations. AB - We consider a sequence of topological torus bifurcations (TTBs) in a nonlinear, quasiperiodic Mathieu equation. The sequence of TTBs and an ensuing transition to chaos are observed by computing the principal Lyapunov exponent over a range of the bifurcation parameter. We also consider the effect of the sequence on the power spectrum before and after the transition to chaos. We then describe the topology of the set of knotted tori that are present before the transition to chaos. Following the transition, solutions evolve on strange attractors that have the topology of fractal braids in Poincare sections. We examine the topology of fractal braids and the dynamics of solutions that evolve on them. We end with a brief discussion of the number of TTBs in the cascade that leads to chaos. PMID- 16252983 TI - Multiple returns for some regular and mixing maps. AB - We study the distributions of the number of visits for some noteworthy dynamical systems, considering whether limit laws exist by taking domains that shrink around points of the phase space. It is well known that for highly mixing systems such limit distributions exhibit a Poissonian behavior. We analyze instead a skew integrable map defined on a cylinder that models a shear flow. Since almost all fibers are given by irrational rotations, we at first investigate the distributions of the number of visits for irrational rotations on the circle. In this last case the numerical results strongly suggest the existence of limit laws when the shrinking domain is chosen in a descending chain of renormalization intervals. On the other hand, the numerical analysis performed for the skew map shows that limit distributions exist even if we take domains shrinking in an arbitrary way around a point, and these distributions appear to follow a power law decay of which we propose a theoretical explanation. It is interesting to note that we observe a similar behavior for domains wholly contained in the integrable region of the standard map. We also consider the case of two or more systems coupled together, proving that the distributions of the number of visits for domains intersecting the boundary between different regions are a linear superposition of the distributions characteristic of each region. Using this result we show that the real limit distributions can be hidden by some finite size effects. In particular, when a chaotic and a regular region are glued together, the limit distributions follow a Poisson-like law, but as long as the measure of the shrinking domain is not zero, the polynomial behavior of the regular component dominates for large times. Such an analysis seems helpful to understand the dynamics in the regions where ergodic and regular motions are intertwined, as it may occur for the standard map. Finally, we study the distributions of the number of visits around generic and periodic points of the dissipative Henon map. Although this map is not uniformly hyperbolic, the distributions computed for generic points show a Poissonian behavior, as usually occurs for systems with highly mixing dynamics, whereas for periodic points the distributions follow a different law that is obtained from the statistics of first return times by assuming that subsequent returns are independent. These results are consistent with a possible rapid decay of the correlations for the Henon map. PMID- 16252984 TI - Chaos control and synchronization in Bragg acousto-optic bistable systems driven by a separate chaotic system. AB - In this paper we propose a new scheme to achieve chaos control and synchronization in Bragg acousto-optic bistable systems. In the scheme, we use the output of one system to drive two identical chaotic systems. Using the maximal conditional Lyapunov exponent (MCLE) as the criterion, we analyze the conditions for realizing chaos synchronization. Numerical calculation shows that the two identical systems in chaos with negative MCLEs and driven by a chaotic system can go into chaotic synchronization whether or not they were in chaos initially. The two systems can go into different periodic states from chaos following an inverse period-doubling bifurcation route as well when driven by a periodic system. PMID- 16252985 TI - Dissipative-Hamiltonian decomposition of smooth vector fields based on symmetries. AB - We propose a method to decompose a smooth vector field into conservative and dissipative components. The procedure is based on the identification of the kernel of a linear operator associated with a given Hamiltonian combined with the use of Lie transformations for vector fields. Moreover, under certain conditions the nonconservative part of the splitting can be dropped at a given order of the transformation, obtaining after truncation, a Hamilton vector field. The technique is illustrated through the application to the motion of a particle subject to the potential of a champagne bottle plus a small friction. PMID- 16252986 TI - Basic structures of the Shilnikov homoclinic bifurcation scenario. AB - We find numerically small scale basic structures of homoclinic bifurcation curves in the parameter space of the Chua circuit. The distribution of these basic structures in the parameter space and their geometrical properties constitute a complete homoclinic bifurcation scenario of this system. Furthermore, these structures and the scenario are theoretically demonstrated to be generic to a large class of dynamical systems that presents, as the Chua circuit, Shilnikov homoclinic orbits. We classify the complexity of primary and subsidiary homoclinic orbits by their order given by the number of their returning loops. Our results confirm previous predictions of structures of homoclinic bifurcation curves and extend this study to high order primary orbits. Furthermore, we identify accumulations of bifurcation curves of subsidiary homoclinic orbits into bifurcation curves of both primary and subsidiary orbits. PMID- 16252987 TI - Reentry wave formation in excitable media with stochastically generated inhomogeneities. AB - Clinical research shows that the frequency of arrhythmia events depends on the number and area of the border zones of infarct scars. We investigate the possibility that arrhythmia is initiated by reentry waves generated by the inhomogeneity of conduction velocity at the border zone. The interaction of a plane wave with a spatially extended inhomogeneity is simulated in the FitzHugh- Nagumo model. The inhomogeneity is introduced into the model by modifying the spatial dependence of the diffusion coefficient in a stochastic manner. This results in a rich variety of spatial distributions of conductivity. A plane wave propagating through such a system may break up on the regions with low conductivity and produce numerous spiral waves. The frequency of reentry wave formation is studied as a function of the parameters of the inhomogeneity generation algorithm. Three main scenarios of reentry wave formation were found: unidirectional block, main wave-wavelet collision, and wave break up during collision, on a region in which a conduction velocity gradient occurs. These scenarios are likely candidates for the mechanisms of arrhythmia initiation in a damaged tissue, e.g., the border zone of an infarct scar. PMID- 16252988 TI - The semiclassical regime of the chaotic quantum-classical transition. AB - An analysis of the semiclassical regime of the quantum-classical transition is given for open, bounded, one-dimensional chaotic dynamical systems. Environmental fluctuations-characteristic of all realistic dynamical systems-suppress the development of a fine structure in classical phase space and damp nonlocal contributions to the semiclassical Wigner function, which would otherwise invalidate the approximation. This dual regularization of the singular nature of the semiclassical limit is demonstrated by a numerical investigation of the chaotic Duffing oscillator. PMID- 16252989 TI - n-body dynamics of stabilized vector solitons. AB - In this work we study the interactions between stabilized Townes solitons. By means of effective Lagrangian methods, we have found that the interactions between these solitons are governed by central forces, in a first approximation. In our numerical simulations we describe different types of orbits, deflections, trapping, and soliton splitting. Splitting phenomena are also described by finite dimensional reduced models. All these effects could be used for potential applications of stabilized solitons. PMID- 16252990 TI - Scaling properties for a classical particle in a time-dependent potential well. AB - Some scaling properties for a classical particle interacting with a time dependent square-well potential are studied. The corresponding dynamics is obtained by use of a two-dimensional nonlinear area-preserving map. We describe dynamics within the chaotic sea by use of a scaling function for the variance of the average energy, thereby demonstrating that the critical exponents are connected by an analytic relationship. PMID- 16252991 TI - Dynamic chaos and stability of a weakly open Bose-Einstein condensate in a double well trap. AB - We investigate the dynamics of a weakly open Bose-Einstein condensate with attractive interaction in a magneto-optical double-well trap. A set of time dependent ordinary differential equations describing the complex dynamics are derived by using a two-mode approximation. The stability of the stationary solution is analyzed and some stability regions on the parameter space are displayed. In the symmetric well case, the numerical calculations reveal that by adjusting the feeding from the nonequilibrium thermal cloud or the two-body dissipation rate, the system could transit among the periodic motions, chaotic self-trapping states of the Lorenz model, and the steady states with the zero relative atomic population or with the macroscopic quantum self-trapping (MQST). In the asymmetric well case, we find the periodic orbit being a stable two-sided limited cycle with MQST. The results are in good agreement with that of the direct numerical simulations to the Gross-Pitaevskii equation. PMID- 16252993 TI - Dynamics of self-adjusting systems with noise. AB - We study several self-adjusting systems with noise. In our analytical and numerical studies, we find that the dynamics of the self-adjusting parameter can be accurately described with a rescaled diffusion equation. We find that adaptation to the edge of chaos, a feature previously ascribed to self-adjusting systems, is only a long-lived transient when noise is present in the system. In addition, using analytical, numerical, and experimental methods, we find that noise can cause chaotic outbreaks where the parameter reenters the chaotic regime and the system dynamics become chaotic. We find that these chaotic outbreaks have a power law distribution in length. PMID- 16252992 TI - Real-time nonlinear feedback control of pattern formation in (bio)chemical reaction-diffusion processes: a model study. AB - Theoretical and experimental studies related to manipulation of pattern formation in self-organizing reaction-diffusion processes by appropriate control stimuli become increasingly important both in chemical engineering and cellular biochemistry. In a model study, we demonstrate here exemplarily the application of an efficient nonlinear model predictive control (NMPC) algorithm to real-time optimal feedback control of pattern formation in a bacterial chemotaxis system modeled by nonlinear partial differential equations. The corresponding drift diffusion model type is representative for many (bio)chemical systems involving nonlinear reaction dynamics and nonlinear diffusion. We show how the computed optimal feedback control strategy exploits the system inherent physical property of wave propagation to achieve desired control aims. We discuss various applications of our approach to optimal control of spatiotemporal dynamics. PMID- 16252994 TI - Studying vocal fold vibrations in Parkinson's disease with a nonlinear model. AB - A nonlinear model is applied to study pathologic vocal vibratory characteristics and voice treatments of Parkinson's disease. We find that a number of pathologic vocal characteristics commonly observed in Parkinson's disease, including reduced vibratory intensity, incomplete vocal closure, increased phonation threshold pressure, glottal tremor, subharmonics, and chaotic vocal fold vibrations, can be studied with this nonlinear model. We also find that two kinds of clinical voice treatments for Parkinson's disease, including respiratory effort treatment and Lee Silverman voice treatment can be studied with this computer model. Results suggest that respiratory effort treatment, in which subglottal pressure is increased, might aid in enhancing vibratory intensity, improving glottal closure, and avoiding vibratory irregularity. However, the Lee Silverman voice treatment, in which both subglottal pressure and vocal fold adduction are increased, might be better than respiratory effort treatment. Increasing vocal fold thickness would be further helpful to improve these pathologic characteristics. The model studies show consistencies with clinical observations. Computer models may be of value in understanding the dynamic mechanism of disordered voices and studying voice treatment effects in Parkinson's disease. PMID- 16252995 TI - Stability of functions in Boolean models of gene regulatory networks. AB - Boolean networks are used to model large nonlinear systems such as gene regulatory networks. We will present results that can be used to understand how the choice of functions affects the network dynamics. The so called bias-map and its fixed points depict much of the function's dynamical role in the network. We define the concept of stabilizing functions and show that many Post and canalizing functions are also stabilizing functions. Boolean networks constructed using the same type of stabilizing functions are always stable regardless of the average in-degree of network functions. We derive the number of all stabilizing functions and find it to be much larger than the number of Post and canalizing functions. We also discuss the implementation of functions and apply the presented results to biological data that give an approximation of the distribution of regulatory functions in eucaryotic cells. We find that the obtained theoretical results on the number of active genes are biologically plausible. Finally, based on the presented results, we discuss why canalizing and Post regulatory functions seem to be common in cells. PMID- 16252996 TI - Solitons in nonintegrable systems. AB - We introduce the concept of this special focus issue on solitons in nonintegrable systems. A brief overview of some recent developments is provided, and the various contributions are described. The topics covered in this focus issue are the modulation of solitons, bores, and shocks, the dynamical evolution of solitary waves, and existence and stability of solitary waves and embedded solitons. PMID- 16252997 TI - Analytic model for a weakly dissipative shallow-water undular bore. AB - We use the integrable Kaup-Boussinesq shallow water system, modified by a small viscous term, to model the formation of an undular bore with a steady profile. The description is made in terms of the corresponding integrable Whitham system, also appropriately modified by viscosity. This is derived in Riemann variables using a modified finite-gap integration technique for the Ablowitz-Kaup-Newell Segur (AKNS) scheme. The Whitham system is then reduced to a simple first-order differential equation which is integrated numerically to obtain an asymptotic profile of the undular bore, with the local oscillatory structure described by the periodic solution of the unperturbed Kaup-Boussinesq system. This solution of the Whitham equations is shown to be consistent with certain jump conditions following directly from conservation laws for the original system. A comparison is made with the recently studied dissipationless case for the same system, where the undular bore is unsteady. PMID- 16252998 TI - Resolution of a shock in hyperbolic systems modified by weak dispersion. AB - We present a way to deal with dispersion-dominated "shock-type" transition in the absence of completely integrable structure for the systems that one may characterize as strictly hyperbolic regularized by a small amount of dispersion. The analysis is performed by assuming that the dispersive shock transition between two different constant states can be modeled by an expansion fan solution of the associated modulation (Whitham) system for the short-wavelength nonlinear oscillations in the transition region (the so-called Gurevich-Pitaevskii problem). We consider both single-wave and bidirectional systems. The main mathematical assumption is that of hyperbolicity of the Whitham system for the solutions of our interest. By using general properties of the Whitham averaging for a certain class of nonlinear dispersive systems and specific features of the Cauchy data prescription on characteristics we derive a set of transition conditions for the dispersive shock, actually bypassing full integration of the modulation equations. Along with the Korteweg-de Vries (KdV) and modified KdV (mKdV) equations as model examples, we consider a nonintegrable system describing fully nonlinear ion-acoustic waves in collisionless plasma. In all cases our transition conditions are in complete agreement with previous analytical and numerical results. PMID- 16252999 TI - Evolution of two-dimensional lump nanosolitons for the Zakharov-Kuznetsov and electromigration equations. AB - The evolution of lump solutions for the Zakharov-Kuznetsov equation and the surface electromigration equation, which describes mass transport along the surface of nanoconductors, is studied. Approximate equations are developed for these equations, these approximate equations including the important effect of the dispersive radiation shed by the lumps as they evolve. The approximate equations show that lump-like initial conditions evolve into lump soliton solutions for both the Zakharov-Kuznetsov equation and the surface electromigration equations. Solutions of the approximate equations, within their range of applicability, are found to be in good agreement with full numerical solutions of the governing equations. The asymptotic and numerical results predict that localized disturbances will always evolve into nanosolitons. Finally, it is found that dispersive radiation plays a more dominant role in the evolution of lumps for the electromigration equations than for the Zakharov Kuznetsov equation. PMID- 16253000 TI - Averaging of nonlinearity-managed pulses. AB - We consider the nonlinear Schrodinger equation with the nonlinearity management which describes Bose-Einstein condensates under Feshbach resonance. By using an averaging theory, we derive the Hamiltonian averaged equation and compare it with other averaging methods developed for this problem. The averaged equation is used for analytical approximations of nonlinearity-managed solitons. PMID- 16253001 TI - Forced solitary waves and fronts past submerged obstacles. AB - Herein, an efficient numerical method is presented to describe the flow of a liquid in an open channel with various types of bottom configurations. The method is developed for steady two-dimensional potential free surface flows. The resulting nonlinear problem is solved numerically by boundary integral equation methods. In addition weakly nonlinear solutions are derived. New solutions which complement those of Dias and Vanden-Broeck [J. Fluid Mech. 59, 93-102 (2004)] are presented. Furthermore some solutions for channel flows past dips in the bottom are discussed. PMID- 16253002 TI - Solitary wave dynamics in shallow water over periodic topography. AB - The problem of long-wave scattering by piecewise-constant periodic topography is studied both for a linear solitary-like wave pulse, and for a weakly nonlinear solitary wave [Korteweg-de Vries (KdV) soliton]. If the characteristic length of the topographic irregularities is larger than the pulse length, the solution of the scattering problem is obtained analytically for a leading wave in the framework of linear shallow-water theory. The wave decrement in the case of the small height of the topographic irregularities is proportional to delta2, where delta is the relative height of the topographic obstacles. An analytical approximate solution is also obtained for the weakly nonlinear problem when the length of the irregularities is larger than the characteristic nonlinear length scale. In this case, the Korteweg-de Vries equation is solved for each piece of constant depth by using the inverse scattering technique; the solutions are matched at each step by using linear shallow-water theory. The weakly nonlinear solitary wave decays more significantly than the linear solitary pulse. Solitary wave dynamics above a random seabed is also discussed, and the results obtained for random topography (including experimental data) are in reasonable agreement with the calculations for piecewise topography. PMID- 16253003 TI - Solitons in a linearly coupled system with separated dispersion and nonlinearity. AB - We introduce a model of dual-core waveguide with the cubic nonlinearity and group velocity dispersion (GVD) confined to different cores, with the linear coupling between them. The model can be realized in terms of photonic-crystal fibers. It opens a way to understand how solitons are sustained by the interplay between the nonlinearity and GVD which are not "mixed" in a single nonlinear Schrodinger (NLS) equation, but are instead separated and mix indirectly, through the linear coupling between the two cores. The spectrum of the system contains two gaps, semi-infinite and finite ones. In the case of anomalous GVD in the dispersive core, the solitons fill the semi-infinite gap, leaving the finite one empty. This soliton family is entirely stable, and is qualitatively similar to the ordinary NLS solitons, although shapes of the soliton's components in the nonlinear and dispersive cores are very different, the latter one being much weaker and broader. In the case of the normal GVD, the situation is completely different: the semi-infinite gap is empty, but the finite one is filled with a family of stable gap solitons featuring a two-tier shape, with a sharp peak on top of a broad "pedestal." This case has no counterpart in the usual NLS model. An extended system, including weak GVD in the nonlinear core, is analyzed too. In either case, when the solitons reside in the semi-infinite or finite gap, they persist if the extra GVD is anomalous, and completely disappear if it is normal. PMID- 16253004 TI - Generation of Bragg solitons through modulation instability in a Bragg grating structure. AB - In this article, we consider the continuous wave (cw) propagation through the nonlinear periodic structure that consists of alternating layers of both positive and negative Kerr coefficients along the propagation direction. We investigate the modulational instability (MI) conditions required for the generation of ultrashort pulses for the nonlinearity management system. We study the occurrence of MI at the top and bottom edges of the photonic band gap (PBG) where the forward and backward propagating waves are strongly coupled because of the presence of the grating structure. We also study the MI when cw is detuned from the edges of the PBG into the anomalous and normal dispersion regimes. In addition, we discuss the existence of gap solitons for the nonlinearity management system in the upper and lower branches of the dispersion curve through the MI gain spectra. We observe the generation of higher order solitons in the nonlinear periodic structure when the input power is increased beyond a certain critical level. Finally, we discuss the generation of higher order Bragg grating solitons through the intensity evolution of the forward and backward propagating fields. PMID- 16253005 TI - Generation, propagation, and breaking of internal solitary waves. AB - Tidal, two-layer flow over topography generates a kink of the interface separating an upstream interfacial elevation from a depression above the topography. Upstream undular bores and solitary waves of large amplitude are generated from the interfacial kink. The waves propagate upstream when the tide turns. Interfacial simulations of this kind of generation process fit with the observations at Knight Inlet in British Columbia, in the Sulu Sea experiment, and undular bores generated by internal tides in the Strait of Gibraltar. Fully nonlinear interfacial computations compare successfully with experimental observations of solitary waves in the laboratory and in the field for wave amplitudes ranging from small to maximal values. The waves exhibit only minor sensitivity to a finite thickness of the pycnocline. Analytical solitary waves are recaptured in the small amplitude limit. Shear-induced breaking appears first in the top part of the pycnocline and is expressed in terms of the Richardson number. Convective breaking in the top part of the water column occurs beyond a threshold amplitude when a pronounced stratification continues all the way to the ocean surface. PMID- 16253006 TI - Internal solitons in laboratory experiments: comparison with theoretical models. AB - Nonlinear internal solitary waves observed in laboratory experiments are discussed from the standpoint of their relation to different soliton theories, from the classical integrable models such as the Korteweg-de Vries, Gardner, Benjamin-Ono, and Joseph-Kubota-Ko-Dobbs equations and their modifications, through the nonintegrable models describing higher-order nonlinear effects, viscosity, rotation, and cylindrical spreading, to the strongly nonlinear models. First, these theoretical models are briefly described and, then, laboratory data and their comparison with the theory are presented. PMID- 16253007 TI - Water waves as a spatial dynamical system; infinite depth case. AB - We review the mathematical results on traveling waves in one or several superposed layers of potential flow, subject to gravity, with or without surface and interfacial tension, where the bottom layer is infinitely deep. The problem is formulated as a "spatial dynamical system," and it is shown that the linearized operator of the resulting reversible system has an essential spectrum filling the real line. We consider three cases where bifurcation occurs. (i) The first case is when, in moving a parameter, two pairs of imaginary eigenvalues merge into one pair of double eigenvalues, and then split into four symmetric complex conjugate eigenvalues. (ii) The second case is when one pair of imaginary eigenvalues meet in 0, and disappear; (iii) the third case is when the phenomenon described in (ii) is superposed to the presence of another pair of imaginary eigenvalues sitting at finite distance from 0. We give a physical example for each case and more specially study the solitary waves and generalized solitary waves, emphasizing the differences, in the methods and in the results, between these cases and the finite depth case. PMID- 16253008 TI - Steady dark solitary waves emerging from wave-generated meanflow: the role of modulation equations. AB - Various classes of steady and unsteady dark solitary waves (DSWs) are known to exist in modulation equations for water waves in finite depth. However, there is a class of steady DSWS of the full water-wave problem which are missed by the classical modulation equations such as the Hasimoto-Ono, Benney-Roskes, and Davey Stewartson. These steady DSWs, recently discovered by Bridges and Donaldson, are pervasive in finite depth, arise through secondary criticality of Stokes gravity waves, and are synchronized with the Stokes wave. In this paper, the role of DSWs in modulation equations for water waves is reappraised. The intrinsic unsteady nature of existing modulation equations filters out some interesting solutions. On the other hand, the geometry of DSWs in modulation equations is very similar to the full water wave problem and these geometrical properties are developed. A model equation is proposed which illustrates the general nature of the emergence of steady DSWs due to wave-generated mean flow coupled to a periodic wave. Although the existing modulation equations are intrinsically unsteady, it is shown that there are also important shortcomings when one wants to use them for stability analysis of DSWs. PMID- 16253009 TI - Bose-Einstein condensates in the presence of a magnetic trap and optical lattice. AB - In this paper we consider solutions of a nonlinear Schrodinger equation with a parabolic and a periodic potential motivated from the dynamics of Bose-Einstein condensates. Our starting point is the corresponding linear problem which we analyze through regular perturbation and homogenization techniques. We then use Lyapunov-Schmidt theory to establish the persistence and bifurcation of the linear states in the presence of attractive and repulsive nonlinear inter particle interactions. Stability of such solutions is also examined and a count is given of the potential real, complex and imaginary eigenvalues with negative Krein signature that such solutions may possess. The results are corroborated with numerical computations. PMID- 16253010 TI - Stability analysis of embedded solitons in the generalized third-order nonlinear Schrodinger equation. AB - We study the generalized third-order nonlinear Schrodinger (NLS) equation which admits a one-parameter family of single-hump embedded solitons. Analyzing the spectrum of the linearization operator near the embedded soliton, we show that there exists a resonance pole in the left half-plane of the spectral parameter, which explains linear stability, rather than nonlinear semistability, of embedded solitons. Using exponentially weighted spaces, we approximate the resonance pole both analytically and numerically. We confirm in a near-integrable asymptotic limit that the resonance pole gives precisely the linear decay rate of parameters of the embedded soliton. Using conserved quantities, we qualitatively characterize the stable dynamics of embedded solitons. PMID- 16253011 TI - Accumulation of embedded solitons in systems with quadratic nonlinearity. AB - Previous numerical studies have revealed the existence of embedded solitons (ESs) in a class of multiwave systems with quadratic nonlinearity, families of which seem to emerge from a critical point in the parameter space, where the zero solution has a fourfold zero eigenvalue. In this paper, the existence of such solutions is studied in a three-wave model. An appropriate rescaling casts the system in a normal form, which is universal for models supporting ESs through quadratic nonlinearities. The normal-form system contains a single irreducible parameter epsilon, and is tantamount to the basic model of type-I second-harmonic generation. An analytical approximation of Wentzel-Kramers-Brillouin type yields an asymptotic formula for the distribution of discrete values of epsilon at which the ESs exist. Comparison with numerical results shows that the asymptotic formula yields an exact value of the scaling index, -65, and a fairly good approximation for the numerical factor in front of the scaling term. PMID- 16253013 TI - The unmet clinical need for new molecular genetic markers in the prognosis and therapeutic management of breast cancer. PMID- 16253012 TI - Prolonged activation of virus-specific CD8+T cells after acute B19 infection. AB - BACKGROUND: Human parvovirus B19 (B19) is a ubiquitous and clinically significant pathogen, causing erythema infectiosum, arthropathy, transient aplastic crisis, and intrauterine fetal death. The phenotype of CD8+ T cells in acute B19 infection has not been studied previously. METHODS AND FINDINGS: The number and phenotype of B19-specific CD8+ T cell responses during and after acute adult infection was studied using HLA-peptide multimeric complexes. Surprisingly, these responses increased in magnitude over the first year post-infection despite resolution of clinical symptoms and control of viraemia, with T cell populations specific for individual epitopes comprising up to 4% of CD8+ T cells. B19 specific T cells developed and maintained an activated CD38+ phenotype, with strong expression of perforin and CD57 and downregulation of CD28 and CD27. These cells possessed strong effector function and intact proliferative capacity. Individuals tested many years after infection exhibited lower frequencies of B19 specific cytotoxic T lymphocytes, typically 0.05%-0.5% of CD8+ T cells, which were perforin, CD38, and CCR7 low. CONCLUSION: This is the first example to our knowledge of an "acute" human viral infection inducing a persistent activated CD8+ T cell response. The likely explanation--analogous to that for cytomegalovirus infection--is that this persistent response is due to low-level antigen exposure. CD8+ T cells may contribute to the long-term control of this significant pathogen and should be considered during vaccine development. PMID- 16253015 TI - What's new in hereditary colorectal cancer? AB - Precancerous polyposes other than classic familial adenomatous polyposis and the condition hereditary nonpolyposis colorectal cancer, or Lynch syndrome, continue to present major diagnostic challenges for the anatomic pathologist. This editorial highlights the practical significance of novel insights and clinical guidelines in the recent literature, as well as in 4 contributions to this edition of the Archives of Pathology & Laboratory Medicine. The first section will address attenuated familial adenomatous polyposis and a newly recognized type of autosomal-recessive adenomatous polyposis associated with the DNA repair gene MYH. The remainder of the editorial discusses the role of the revised Bethesda guidelines in the diagnosis of hereditary nonpolyposis colorectal cancer and concludes with the recently identified serrated pathway syndrome. PMID- 16253016 TI - Testing for defective DNA mismatch repair in colorectal carcinoma: a practical guide. AB - CONTEXT: Significant bench and clinical data have been generated during the last decade regarding DNA mismatch repair in colorectal carcinoma. OBJECTIVES: To review clinically relevant aspects of defective DNA mismatch repair in colorectal carcinoma and to suggest testing algorithms for identification of these tumors in the sporadic and familial settings. DATA SOURCES: This article is based on literature review and clinical testing experience of more than 2000 patient samples. CONCLUSIONS: Approximately 15% of colorectal carcinomas arise as a result of defective DNA mismatch repair. Ninety percent of these carcinomas are sporadic, arising as a result of methylation of the MLH1 gene promoter, silencing expression. These sporadic carcinomas have improved stage-specific prognosis and can be identified by demonstrating aberrant loss of expression with an MLH1 immunoperoxidase stain. Familial colorectal carcinomas with defective DNA mismatch repair (Lynch syndrome) are due to a germline defect in one of several DNA mismatch repair genes. The familial carcinomas are best identified with a combination of immunohistochemistry and molecular microsatellite analysis. This testing facilitates subsequent directed genetic testing of the proband and family members. PMID- 16253018 TI - High-grade pancreatic intraepithelial neoplasia in a patient with familial adenomatous polyposis. AB - Familial adenomatous polyposis (FAP) is caused by mutation of the adenomatous polyposis coli (APC) gene and is characterized by multiple colorectal adenomas and tumors of other organs and sites. A 58-year-old woman with FAP syndrome and previous total colectomy presented for routine follow-up examination. Abdominal ultrasound and subsequent endoscopic evaluation revealed ampullary and duodenal polyps, as well as inhomogeneity of the pancreatic head. A pancreaticoduodenectomy confirmed multiple duodenal adenomas. In addition, high grade pancreatic intraepithelial neoplasia (PanIN-3) was found in the smaller pancreatic ducts. Pancreatic precancerous lesions have only rarely been described in FAP, including 2 pancreatic duct adenomas and 2 intraductal papillary mucinous neoplasms. A review of the world English literature revealed no reports of PanIN 3 in association with FAP. Further studies are required to determine if patients with FAP are at increased risk for pancreatic premalignant lesions. PMID- 16253017 TI - Performance of the revised Bethesda guidelines for identification of colorectal carcinomas with a high level of microsatellite instability. AB - CONTEXT: Criteria for microsatellite instability (MSI) testing to rule out hereditary nonpolyposis colorectal cancer were recently revised and include parameters such as age and specific histologic features that can be identified by the pathologist, triggering reflex MSI testing. OBJECTIVE: To review the performance of the revised Bethesda guidelines to identify MSI-positive colorectal cancers. DESIGN: Seventy-five patients with colorectal cancer were included; 68 patients younger than 50 years and 7 patients between 50 and 60 years were selected based on histopathologic criteria. Microsatellite instability testing with the National Cancer Institute--recommended panel and immunohistochemistry for hMLH1 and hMSH2 were performed. Tumors were classified into microsatellite instability high (MSI-H), low (MSI-L), or stable (MSS) categories. RESULTS: Overall, 17 (23%) of 75 colorectal cancer cases were classified as MSI-H, including 13 patients younger than 50 years and 4 patients between 50 and 60 years. Among the MSI-H tumors, 10 (59%) were characterized by loss of hMLH1 and 6 (35%) were hMSH2 negative. Histologic features suggestive of MSI-H phenotype were present in 80% of MSI-H and 35% of MSS/MSI-L tumors. The number of positive lymph nodes was higher in MSS/MSI-L adenocarcinomas (P = .04). CONCLUSIONS: By selecting for age and histologic features, we detected MSI-H tumors in approximately one quarter of colorectal cancer cases meeting the revised Bethesda guidelines and identified 17 MSI-H cases, whereas only 8 would have been recognized by the prior guidelines. These data indicate that reflex testing requested by pathologists based on the revised Bethesda guidelines increases the detection of MSI-H and potential hereditary nonpolyposis colorectal cancer cases. PMID- 16253019 TI - Attenuated familial adenomatous polyposis: a case report with mixed features and review of genotype-phenotype correlation. AB - Familial adenomatous polyposis represents approximately 1% of all colorectal cancers and is caused by germline mutations in the adenomatous polyposis coli (APC) gene. Most mutations are located within the first 2000 codons, and several mutational hot spots have been identified. The relative location of the mutation may be associated with the number of polyps and partially predicts specific phenotypic expression. Mutations associated with the attenuated phenotype are found predominantly in the 5' region of the gene or in the last third. We describe a patient with a mutation in codon 161 of the APC gene, which displays a phenotype most closely resembling the attenuated form of familial adenomatous polyposis, and review the literature, the implications of this mutation, and the importance of the molecular testing in the proper and more complete characterization of these patients. Differences in the APC mutation sites alone cannot completely account for intrafamilial and interfamilial variation in the polyposis phenotypes. PMID- 16253020 TI - What's new in the pleura? PMID- 16253021 TI - Immunohistochemical diagnosis of epithelioid mesothelioma: an update. AB - OBJECTIVE: Several new immunohistochemical markers that can assist in the diagnosis of mesotheliomas have been recognized recently. This article reviews the current information available on these markers and also provides a practical approach to the immunohistochemical diagnosis of epithelioid mesotheliomas. DATA SOURCES: Current literature concerning immunohistochemical markers for epithelioid mesotheliomas was collected and reviewed. STUDY SELECTION: Literature emphasizing immunohistochemical diagnosis of epithelioid mesotheliomas was selected. DATA EXTRACTION: Data deemed helpful to the general surgical pathologist for the diagnosis of epithelioid mesothelioma were included in this review. DATA SYNTHESIS: Markers identified as potentially useful in the diagnosis of epithelioid mesothelioma include positive markers (namely, calretinin, keratin 5/6, D2-40, podoplanin, mesothelin, and Wilms tumor 1 protein [WT1]) and negative markers (namely, carcinoembryonic antigen, MOC-31, B72.3, and Ber-EP4). Thyroid transcription factor 1 (TTF-1) can assist in determining the lung origin of a carcinoma, and renal cell carcinoma marker (RCC Ma) may help establish its renal origin. CONCLUSIONS: D2-40 and podoplanin are the 2 most recently recognized markers that have been found to be useful in the diagnosis of epithelioid mesotheliomas. Since D2-40 and podoplanin appear to be highly sensitive and specific for epithelioid mesotheliomas, either may be considered for inclusion in the battery of antibodies currently recommended for distinguishing epithelioid mesotheliomas from metastatic carcinomas. However, it should be kept in mind that their utility has not yet been fully determined in routine diagnostic work. PMID- 16253022 TI - Recognition of histopathologic patterns of diffuse malignant mesothelioma in differential diagnosis of pleural biopsies. AB - CONTEXT: Diffuse malignant mesothelioma is generally divided into 3 basic histologic types--epithelial, biphasic, or sarcomatous. However, a great diversity of histologic patterns exists within these 3 basic histologic types. DESIGN: The diverse histologic patterns of diffuse malignant mesothelioma are briefly described, including infrequently encountered patterns. RESULTS: The histologic patterns of diffuse malignant mesothelioma provide a wide variation of features for the pathologist to consider in the differential diagnosis of a pleural biopsy. CONCLUSION: Pathologists should be aware of the varied histologic patterns of diffuse malignant mesothelioma when evaluating pleural biopsies. PMID- 16253023 TI - Differential diagnosis of benign and malignant mesothelial proliferations on pleural biopsies. AB - CONTEXT: Although much of the pathology literature focuses on differential diagnosis of diffuse malignant mesothelioma from other types of cancer, the primary diagnostic challenge facing the pathologist is often whether a mesothelial proliferation on a pleural biopsy represents a malignancy or a benign reactive hyperplasia. DESIGN: Based on previous medical publications, extensive personal consultations, and experience on the United States-Canadian Mesothelioma Reference Panel and the International Mesothelioma Panel, salient information was determined about interpretation of benign versus malignant mesothelial proliferations on pleural biopsies. RESULTS: Differentiation of benign reactive mesothelial hyperplasia from diffuse malignant mesothelioma is often difficult. Benign reactive mesothelial hyperplasia may mimic many features ordinarily associated with malignancy, and diffuse malignant mesothelioma may be cytologically bland. Entrapment of benign reactive mesothelial cells within organizing pleuritis may mimic tissue invasion. CONCLUSIONS: Various histologic clues favor a benign over a malignant mesothelial proliferation and vice versa. Invasion is the most reliable criterion for determining that a mesothelial proliferation is malignant. When there is any doubt that a pleural biopsy represents a malignancy, we recommend a diagnosis of atypical mesothelial proliferation. PMID- 16253024 TI - Review and update of uncommon primary pleural tumors: a practical approach to diagnosis. AB - OBJECTIVE: We address the current classifications and new changes regarding uncommon primary pleural tumors. Primary pleural tumors are divided according to their behavior and are discussed separately as benign tumors, tumors of low malignant potential, and malignant neoplasms. DATA SOURCES: Current literature concerning primary pleural neoplasms was collected and reviewed. STUDY SELECTION: Studies emphasizing clinical, radiological, or pathologic findings of primary pleural neoplasms were obtained. DATA EXTRACTION: Data deemed helpful to the general surgical pathologist when confronted with an uncommon primary pleural tumor was included in this review. DATA SYNTHESIS: Tumors are discussed in 3 broad categories: (1) benign, (2) low malignant potential, and (3) malignant. A practical approach to the diagnosis of these neoplasms in surgical pathology specimens is offered. The differential diagnosis, including metastatic pleural neoplasms, is also briefly addressed. CONCLUSIONS: Uncommon primary pleural neoplasms may mimic each other, as well as mimic metastatic cancers to the pleura and diffuse malignant mesothelioma. Correct diagnosis is important because of different prognosis and treatment implications for the various neoplasms. PMID- 16253025 TI - Nasal chondromesenchymal hamartoma in older children and adults: series and immunohistochemical analysis. AB - CONTEXT: Nasal chondromesenchymal hamartoma is a benign mass lesion of the nasal cavity predominantly described in young infants. These unusual lesions are composed of a proliferation of mesenchymal and cartilaginous elements. Their pathogenesis is unknown, but they may be derived from embryologic rests. To our knowledge, only 1 case in an older child has been reported, and no cases have been reported in adults. OBJECTIVE: To report 4 cases of nasal chondromesenchymal hamartoma occurring in older children and adults, including immunohistochemical analysis of these unusual lesions. DESIGN: Cases identified from our archives were examined to confirm the diagnosis of nasal chondromesenchymal hamartoma. Immunohistochemical analysis was performed using a panel of antibodies (epithelial membrane antigen, smooth muscle actin, all muscle actin, cytokeratin, S100, and KP1) to evaluate for epithelial, smooth muscle, neural, chondroid, and histiocytic differentiation. RESULTS: Four cases of nasal chondromesenchymal hamartoma in patients of 11, 69, 17, and 25 years of age demonstrated histologic evidence of mesenchymal and cartilaginous elements underlying a chronically inflamed respiratory mucosa. Bony and adipose elements and rare glandular elements were interspersed. Cartilaginous elements stained strongly with S100, whereas mesenchymal regions showed variable and weaker staining. Smooth muscle differentiation was seen primarily in the mesenchymal areas. Epithelial membrane antigen was focally positive in all cases. CONCLUSIONS: Nasal chondromesenchymal hamartomas can rarely occur in the older child and adult. Mesenchymal areas show both myofibroblastic and cartilaginous differentiation. We speculate that inflammation or a recapitulation of developmental signals may be components in the pathogenesis of these lesions. PMID- 16253026 TI - Cell proliferation and inflammation on biopsy samples with multifocal atrophic gastritis before and 1 year after Helicobacter pylori eradication. AB - CONTEXT: Results of clinical trials that have assessed whether gastric cancer is preventable with Helicobacter pylori eradication therapy remain inconclusive. These trials have used atrophy, intestinal metaplasia, and dysplasia as histopathologic end points that reflect possible preneoplastic lesions. Trial results would be more compelling if cell proliferation and inflammatory markers improved simultaneously with histopathologic lesions. OBJECTIVE: To study the presence of cell proliferation markers and type of inflammatory cells in biopsy specimens with gastritis, atrophy, and intestinal metaplasia before and 1 year after H pylori therapy and to determine if immunohistochemistry can be used to study these. DESIGN: We evaluated 12 subjects with gastritis and 16 with gastritis and multiple foci of atrophy and intestinal metaplasia by using immunohistochemical assays for tumor suppressor protein p53, proliferation marker Ki-67, cell cycle regulator cyclin D1, T and B lymphocytes, macrophages, and TUNEL (terminal deoxynucleotide transferase deoxyuridine triphosphate nick end labeling) assay for apoptosis. The biopsy specimens were selected from a randomized clinical trial that studied improvement of histopathologic gastric lesions after H pylori eradication. RESULTS: Groups of surface epithelial cells that expressed p53 and Ki-67 were observed more often in subjects with atrophy and intestinal metaplasia compared with those with gastritis alone. T lymphocytes in the lamina propria were frequently observed 1 year after treatment in subjects with atrophy and intestinal metaplasia. CONCLUSIONS: Immunohistochemical assays for cell proliferation and inflammatory cell markers showed different distribution patterns in these gastric biopsy specimens. The presence of T lymphocytes and groups of cells that expressed proliferation markers in subjects with multiple foci of atrophy and intestinal metaplasia needs further study. PMID- 16253027 TI - The effects of diseases, drugs, and chemicals on the creativity and productivity of famous sculptors, classic painters, classic music composers, and authors. AB - CONTEXT: Many myths, theories, and speculations exist as to the exact etiology of the diseases, drugs, and chemicals that affected the creativity and productivity of famous sculptors, classic painters, classic music composers, and authors. OBJECTIVE: To emphasize the importance of a modern clinical chemistry laboratory and hematology coagulation laboratory in interpreting the basis for the creativity and productivity of various artists. DESIGN: This investigation analyzed the lives of famous artists, including classical sculptor Benvenuto Cellini; classical sculptor and painter Michelangelo Buonarroti; classic painters Ivar Arosenius, Edvard Munch, and Vincent Van Gogh; classic music composer Louis Hector Berlioz; and English essayist Thomas De Quincey. The analysis includes their illnesses, their famous artistic works, and the modern clinical chemistry, toxicology, and hematology coagulation tests that would have been important in the diagnosis and treatment of their diseases. CONCLUSIONS: The associations between illness and art may be close and many because of both the actual physical limitations of the artists and their mental adaptation to disease. Although they were ill, many continued to be productive. If modern clinical chemistry, toxicology, and hematology coagulation laboratories had existed during the lifetimes of these various well-known individuals, clinical laboratories might have unraveled the mysteries of their afflictions. The illnesses these people endured probably could have been ascertained and perhaps treated. Diseases, drugs, and chemicals may have influenced their creativity and productivity. PMID- 16253028 TI - Epithelial displacement in breast lesions: a papillary phenomenon. AB - CONTEXT: Displacement of epithelial cells (DE) in the breast may occur after various types of needling procedures. OBJECTIVE: To determine if specific lesions or entities in the breast are more prone to displacement than others. DESIGN: A review of our computer files from January 1994 to June 2004 yielded 53 cases with DE. Clinical information, including the age of the patient, specific reason for the biopsy, and type of biopsy, was gathered. Histologic review of all hematoxylin-eosin-stained slides (core biopsies and excisions) was performed. RESULTS: Needling procedures included 1 or more of the following: ultrasound guided core biopsy (24 cases), mammotome core biopsies (16), fine-needle aspiration (8), anesthetic injection (3), suture placement (5), and wire localization (1). Procedures were performed in order to investigate a mass (34 cases), calcifications (15), both (3), or nipple discharge (1). The time from needling to surgical procedure yielding a specimen with DE ranged from minutes to 47 days. Displacement of epithelial cells occurred in the following sites: biopsy tract (42 cases), lymphatic channels (5), both biopsy tract and lymphatic channels (4), and breast stroma (2). The diagnoses included intraductal papilloma (6 cases) and intraductal carcinoma (DCIS) (45; 15 with invasive carcinoma). The remaining 2 cases were invasive carcinoma (colloid and papillary types) devoid of DCIS. Of the DCIS cases, either pure or with invasive carcinoma, the pattern was micropapillary in 23, intraductal papilloma involved by DCIS in 32, and both features in 12. The remaining 2 cases of DCIS included comedo DCIS and cribriform DCIS involving a cyst. CONCLUSIONS: With the exception of 3 cases, DE was associated with 1 or more underlying papillary lesions, including pure intraductal papilloma, DCIS involving intraductal papilloma, micropapillary DCIS, and invasive carcinoma. Other etiologies included mucinous carcinoma and cystic lesions, with only 1 case in which a mechanism for DE could not be postulated. PMID- 16253029 TI - The Italian quality control scheme for crossmatching procedures and HLA sera screening: the 2002 pilot study. AB - CONTEXT: The first national quality control (QC) program of histocompatibility serum testing was performed in Italy in 2002. OBJECTIVE: To monitor the performance of HLA typing laboratories while meeting the accreditation requirements of the European Federation for Immunogenetics (EFI), which require HLA typing laboratories to participate in external QC of their crossmatch and antibody analyses. DESIGN: The Turin Transplant Immunology Service was asked to organize a QC survey of 17 HLA typing laboratories in Italy. Each laboratory received 12 serum specimens and 6 blood samples and was required to perform 36 crossmatches and 12 serum antibody specificity determinations. SETTINGS: Data of participating centers were compared to establish whether EFI requirements were satisfied. RESULTS: In crossmatch analysis, the results of 32 of 36 crossmatches reached the 75% consensus target, with all the participating laboratories meeting the standards of the EFI. In antibody analysis, only 7 of 17 laboratories met the EFI standards. CONCLUSION: The first Italian QC program shows that the participating laboratories obtained consistent results in crossmatching, whereas the results were less satisfactory in the determination of serum antibody specificity, where consensus was reached only with monospecific sera and antibody negative samples. PMID- 16253030 TI - Death certification errors at an academic institution. AB - CONTEXT: The correctly completed death certificate provides invaluable personal, epidemiologic, and legal information and should be thorough and accurate. Death certification errors are common and range from minor to severe. OBJECTIVE: To determine the frequency and type of errors by nonpathologist physicians at a university-affiliated medical center. DESIGN: Fifty random patients were identified who died at this academic medical center between January 2002 and December 2003 and did not undergo an autopsy. From medical chart review, clinical summaries were produced. Two pathologists used these summaries to create mock death certificates. The original and mock death certificates were then compared to identify errors in the original certificate. Errors were graded on a I to IV scale, with grade IV being the most severe. RESULTS: Of the 50 death certificates reviewed, grade I, II, and III errors were noted in 72%, 32%, and 30%, respectively. Seventeen certificates (34%) had grade IV errors (wrong cause or manner of death). Multiple errors were identified in 82% of the death certificates reviewed. CONCLUSIONS: The rate of major (grade IV) death certification errors at this academic setting is high and is consistent with major error rates reported by other academic institutions. We attribute errors to house staff inexperience, fatigue, time constraints, unfamiliarity with the deceased, and perceived lack of importance of the death certificate. To counter these factors, we recommend a multifaceted approach, including an annual course in death certification and discussion of the death certificate for each deceased patient during physician rounds. These measures should result in increased accuracy of this important document. PMID- 16253031 TI - Hepatic follicular dendritic cell sarcoma without Epstein-Barr virus expression. AB - Follicular dendritic cell sarcoma of the liver is an uncommon pathologic entity, and only 5 cases have been reported previously. Herein, we report the first case, to our knowledge, of hepatic follicular dendritic cell sarcoma without evidence of Epstein-Barr virus infection. The patient is an elderly man who was found to have an incidental liver mass and then developed weight loss and fever. The diagnosis was based on the typical morphologic appearance of spindle cell proliferation associated with a brisk lymphoplasmacytic infiltrate and strong immunoreactivity to CD21 and CD35. Based on our experience and a review of the published reports, we summarize the clinical and pathologic features of hepatic follicular dendritic cell sarcoma and its surgical management. PMID- 16253032 TI - Clear cell myomelanocytic tumor (PEComa) of the duodenum in a child with a history of neuroblastoma. AB - We report herein a case of digestive clear cell myomelanocytic tumor (PEComa) that is unique in its location and presentation. The lesion, located in the duodenal wall, was diagnosed in a child with a history of cervical neuroblastoma that was in remission after surgical resection and chemotherapy. The diagnosis was obtained by examination of a biopsy specimen taken during laparoscopy. The decision was made to perform surgical resection. Examination of the surgical specimen confirmed the diagnosis of PEComa. No metastasis was found. After 2 years of follow-up, the patient is alive, without evidence of metastasis or recurrence. This case highlights the distinctive characteristics of the cells in PEComa, recognizable even on limited biopsy material. It also suggests a possible association between PEComa and neuroblastoma, 2 unusual tumors that belong to the spectrum of lesions known to occur in patients with tuberous sclerosis and that may share a possible common pathogenetic mechanism. PMID- 16253033 TI - Nonsecretory variant of immunoproliferative small intestinal disease: a case report with pathologic, immunophenotypic, and molecular findings. AB - We report a case of the nonsecretory variant of immunoproliferative small intestinal disease involving the distal small bowel and the mesenteric and retroperitoneal lymph nodes in a 19-year-old woman from Mexico. This variant extranodal marginal zone B-cell lymphoma appeared similar in the different sites of involvement, with more interspersed large cells and greater plasmacytic differentiation present in intestinal specimens. Characteristic lymphoepithelial lesions and follicular colonization were seen in intestinal and lymph node sections, respectively. The neoplastic B cells were cytoplasmic immunoglobulin (Ig) A heavy-chain restricted and lacked surface and cytoplasmic light-chain expression by flow cytometric analysis. Serum and urine protein electrophoresis/immunofixation revealed hypogammaglobulinemia with no paraprotein. Molecular studies showed absence of immunoglobulin heavy-chain (IgH) gene rearrangement, with a nonfunctional clonotypic rearrangement of the kappa light-chain gene. This case highlights the role for kappa light-chain gene evaluation in immunoproliferative small intestinal disease, because IgH gene rearrangement analysis is often negative. PMID- 16253034 TI - Internal auditory canal mass in a 46-year-old woman. PMID- 16253035 TI - A mass of the right lacrimal sac in a 53-year-old man. PMID- 16253036 TI - A rare tricuspid anomaly. PMID- 16253037 TI - A 62-year-old woman with a suspected thyroid nodule. PMID- 16253038 TI - A 16-year-old girl with hypochromic microcytic anemia. PMID- 16253039 TI - A rare prostatic tumor in a 48-year-old man. PMID- 16253040 TI - A 57-year-old man with a dermal mass in his left ring finger. PMID- 16253041 TI - Without scientific integrity, there can be no evidence base. PMID- 16253042 TI - Beyond disclosure: seeking forgiveness. PMID- 16253043 TI - Scapular muscle tests in subjects with shoulder pain and functional loss: reliability and construct validity. AB - BACKGROUND AND PURPOSE: Scapular muscle performance evaluated with a handheld dynamometer (HHD) has been investigated only in people without shoulder dysfunction for test-retest reliability of data obtained with a single scapular muscle test. The purpose of this study was to assess the reliability, error, and validity of data obtained with an HHD for 4 scapular muscle tests in subjects with shoulder pain and functional loss. SUBJECTS AND METHODS: Subjects (N=40) with shoulder pain and functional loss were tested by measuring the kilograms applied with an HHD during 3 trials for muscle tests for the lower trapezius, upper trapezius, middle trapezius, and serratus anterior muscles. Concurrently, surface electromyography (sEMG) data were collected for the 4 muscles. The same procedures were performed 24 to 72 hours after the initial testing by the same tester. Muscle tests were performed 3 times, and the results were averaged for data analysis. RESULTS: Intraclass correlation coefficients for intratester reliability of measurements of isometric force obtained using an HHD ranged from .89 to .96. The standard error of the measure (90% confidence interval [CI]) ranged from 1.3 to 2.7 kg; the minimal detectable change (90% CI) ranged from 1.8 to 3.6 kg. Construct validity assessment, done by comparing the amounts of isometric muscle activity (sEMG) for each muscle across the 4 muscle tests, revealed that the muscle activity of the upper trapezius and lower trapezius muscles was highest during their respective tests. Conversely, the isometric muscle activity of the middle trapezius and serratus anterior muscles was not highest during their respective tests. DISCUSSION AND CONCLUSION: In people with shoulder pain and functional loss, the intrarater reliability and error over 1 to 3 days were established using an HHD for measurement of isometric force for the assessment of scapular muscle performance. Error values can be used to make decisions regarding individual patients. Construct validity was established for the lower and upper trapezius muscle tests; therefore, these tests are advocated for use. However, construct validity was not demonstrated for the serratus anterior and middle trapezius muscle tests as performed in this study. Further investigation of these muscle tests is warranted. PMID- 16253044 TI - Interventions that increase or decrease the likelihood of a meaningful improvement in physical health in patients with sciatica. AB - BACKGROUND AND PURPOSE: The purpose of our study was to determine whether physical therapy interventions predicted meaningful short-term improvement in physical health for patients diagnosed with sciatica. SUBJECTS: We examined data from 1,804 patients (age: mean=52.1 years, SD=15.6 years; 65.7% female, 34.3% male) who had been diagnosed with sciatica and who had completed an episode of outpatient physical therapy. METHODS: Principal components factor analysis was used to define intervention categories from specific treatments applied during the plan of care. A nested-model logistic regression analysis identified intervention categories that predicted meaningful improvement in physical health. Meaningful improvement was defined as a change of 14 or more points on the Physical Component Scale-12 (PCS-12) summary score. RESULTS: Twenty-six percent (n=473) of patients had a meaningful improvement in physical health. Improvement was more likely in patients receiving joint mobility interventions (odds ratio [OR]=2.5, 95% confidence interval [CI]=1.5-4.4) or general exercise (OR=1.5, 95% CI=1.2-2.0). Patients who received spasm reduction interventions were less likely to improve (OR=0.77, 95% CI=0.60-0.98). DISCUSSION AND CONCLUSION: Physical therapists should emphasize the use of joint mobility interventions and exercise when treating patients with sciatica, whereas interventions for spasm reduction should be avoided. PMID- 16253045 TI - Physical therapists' use of interventions with high evidence of effectiveness in the management of a hypothetical typical patient with acute low back pain. AB - BACKGROUND AND PURPOSE: Evidence-based practice aims to improve patient care and service delivery, particularly in the management of individuals with low back pain (LBP), the largest client group seen by outpatient physical therapists. The purpose of this study was to determine the prevalence of use of interventions with evidence of effectiveness in the management of acute nonspecific LBP by physical therapists. SUBJECTS: A multicenter cross-sectional study was conducted on 100 physical therapists working with patients with LBP. METHODS: Using a telephone-administered interview, therapists described their current and desired treatment practices for a typical case of LBP. Each intervention reported was coded according to its evidence of effectiveness (strong, moderate, limited, or none). Information on clinician, workplace, and client characteristics also was obtained. RESULTS: The prevalence of use of interventions with strong or moderate evidence of effectiveness was 68%. However, 90% to 96% of therapists also used interventions for which research evidence was limited or absent. Users of interventions with high evidence of effectiveness, as compared with nonusers, had graduated more recently and had taken a higher number of postgraduate clinical courses. DISCUSSION AND CONCLUSION: Although most therapists use interventions with high evidence of effectiveness, much of their patient time is spent on interventions that are not well reported in the literature. The results indicate the need for improvement in the quality of clinical research as well as its dissemination and implementation in a way that is appealing to therapists, such as through practice-related courses. PMID- 16253046 TI - Management of work-related low back pain: a population-based survey of physical therapists. AB - BACKGROUND AND PURPOSE: Physical therapy often is used in the management of work related low back pain (LBP). Little information, however, is known about the types of interventions used by physical therapists in the management of this condition. The objective of this study was to describe the interventions used by physical therapists in the treatment of workers with acute or subacute LBP, with or without radiating pain below the knee. SUBJECTS: Clinical management questionnaires for workers without and with radiating pain were returned by 190 and 139 physical therapists, respectively. METHODS: For each treatment session, therapists recorded treatment objectives, interventions, and education provided to 2 workers with LBP, 1 with radiating pain and 1 without radiating pain. RESULTS: The majority of physical therapists used stretching and strengthening exercises, spinal mobilization, soft tissue mobilization and massage, manual traction, posture correction, interferential current, ultrasound, heat, and functional activities education. With radiating pain, the majority of the therapists also used cold and the McKenzie approach. Treatment objectives pursued by the majority of the therapists were decrease of pain, increase of range of motion, increase of muscle strength (force-generating capacity of muscle), decrease of muscle tension, and worker education. DISCUSSION AND CONCLUSION: Physical therapists use an array of interventions with workers with LBP. The effectiveness of most interventions reported has not been well studied. PMID- 16253047 TI - A fitness program for children with disabilities. AB - BACKGROUND AND PURPOSE: This case report describes a fitness program for children with disabilities and provides preliminary information about the safety and feasibility of the program. CASE DESCRIPTION: Nine children, 5 to 9 years of age with physical or other developmental disabilities, participated in a 14-week group exercise program held 2 times per week followed by a 12-week home exercise program. Energy expenditure index, leg strength (force-generating capacity of muscle), functional skills, fitness, self-perception, and safety were measured before intervention, after the group exercise program, and again after the home exercise program. OUTCOMES: No injuries occurred, and improvements in many of the outcome measures were observed. More improvements were observed after the group exercise program than after the home program, and adherence was better during the group exercise program. DISCUSSION: This case report demonstrates that a group exercise program of strength and endurance training may be a safe and feasible option for children with disabilities. Further research is needed to evaluate the effectiveness of a group fitness program and optimal training parameters. PMID- 16253049 TI - Progressive resistance exercise in physical therapy: a summary of systematic reviews. AB - Progressive resistance exercise (PRE) is a method of increasing the ability of muscles to generate force. However, the effectiveness and safety of PRE for clients of physical therapists are not well known. The purpose of this article is to review the evidence on positive and negative effects of PRE as a physical therapy intervention. Electronic databases were searched for systematic reviews on PRE and any relevant randomized trials published after the last available review. The search yielded 18 systematic reviews under major areas of physical therapy: cardiopulmonary, musculoskeletal, neuromuscular, and gerontology. Across conditions, PRE was shown to improve the ability to generate force, with moderate to large effect sizes that may carry over into an improved ability to perform daily activities. Further research is needed to determine the potential negative effects of PRE, how to maximize carryover into everyday activities, and what effect, if any, PRE has on societal participation. PMID- 16253048 TI - Patellofemoral pain and asymmetrical hip rotation. AB - BACKGROUND AND PURPOSE: Patellofemoral joint problems are the most common overuse injury of the lower extremity, and altered femoral or hip rotation may play a role in patellofemoral pain. The purpose of this case report is to describe the evaluation of and intervention for a patient with asymmetrical hip rotation and patellofemoral pain. CASE DESCRIPTION: The patient was a 15-year-old girl with an 8-month history of anterior right knee pain, without known trauma or injury. Prior to intervention, her score on the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) was 24%. Right hip medial (internal) rotation was less than left hip medial rotation, and manual muscle testing showed weakness of the right hip internal rotator and abductor muscles. The intervention was aimed at increasing right hip medial rotation, improving right hip muscle strength (eg, the muscle force exerted by a muscle or a group of muscles to overcome a resistance), and eliminating anterior right knee pain. OUTCOMES: After 6 visits (14 days), passive left and right hip medial rotations were symmetrical, and her right hip internal rotator and abductor muscle grades were Good plus. Her WOMAC score was 0%. DISCUSSION: The patient had right patellofemoral pain and an uncommon pattern of asymmetrical hip rotation, with diminished hip medial rotation and excessive hip lateral (external) rotation on the right side. The patient's outcomes suggest that femoral or hip joint asymmetry may be related to patellofemoral joint pain. PMID- 16253050 TI - Thirty-Sixth Mary McMillan Lecture: Never satisfied. PMID- 16253051 TI - 2005 APTA Presidential Address: "For the sake of our patients, it is the right thing to do". PMID- 16253052 TI - Twenty-five years of shockwave lithotripsy: back to the future? PMID- 16253053 TI - Laparoscopy. AB - Urologists around the world are doing more laparoscopy, with renal surgery being the most common procedure. Almost a third devote more than 20% of their practices to laparoscopy. Among those who do not use laparoscopy, lack of training and equipment are the most common reasons. PMID- 16253054 TI - Ureteral metal stents: a tale or a tool? AB - There are four types of ureteral metal stents: self expandable, balloon expandable, covered, and thermoexpandable shape-memory. Insertion of metal stents requires expertise with transurethral and percutaneous techniques. The stricture is traversed with the aid of a guidewire via a percutaneous nephrostomy, and the stenotic segment is dilated using a high-pressure balloon catheter. The stent is then inserted over the guidewire, such that the upper end bypasses the obstruction by at least 3 to 4 cm, while the lower end extends intravesically for 0.5 to 1 cm from the ureteral orifice. If necessary, two or more stents are placed in sequence, overlapping by at least 2 to 3 cm. Metal stents were initially used for the relief of end-stage malignant disease, and their role in the treatment of benign ureteral strictures is still undefined. Patients often complain of abdominal discomfort and mild pain after stent insertion, which soon resolve spontaneously. Hematuria usually stops after a few days and does not necessitate any treatment. Mild urothelial hyperplasia in the stent lumen is common but usually regresses after 4 to 6 weeks. Many authors suggest the use of a double-pigtail catheter for the first 4 to 6 weeks to avoid narrowing of the ureteral lumen. The influence of stents on ureteral peristalsis is a major but poorly documented issue. Encrustation is a significant problem that needs to be addressed. The characteristics of both the patient and the stent influence its likelihood. Migration of coated metal stents was seen in 81% of patients at our center. Virtual endoscopy has recently been introduced as a tool for the follow up of patients with stented ureters. Further design development is necessary to obtain the ideal ureteral metal stent. In a recent study in female pigs, paclitaxel-eluting metal stents engendered less inflammation and hyperplasia of the surrounding tissues. PMID- 16253055 TI - Trends in robotic surgery. AB - Robotic surgery began as a technology-driven innovation but is now becoming a genuine method of improving healthcare effectiveness worldwide. This comprehensive review introduces the current trends, using examples of specific systems to distinguish the various types of robotic surgical devices, from remote handling machines to those performing delicate local interventions. We end by commenting on how to extend existing systems and provide an account of the rapid developments in minimally invasive robotic surgery. PMID- 16253056 TI - A 60-year milestone for Karl Storz company. PMID- 16253057 TI - Controversial cases in endourology. PMID- 16253058 TI - Investigation of acute flank pain: how do practices of U.K.and Irish urologists compare with those of transatlantic and continental European colleagues? AB - BACKGROUND AND PURPOSE: Noncontrast-enhanced spiral CT (NESCT) is more accurate and reliable than intravenous urography (IVU) in diagnosing the cause of acute flank pain. This study aimed to determine the impact of current literature on the choice of imaging modality used to investigate acute flank pain within the UK and the Republic of Ireland. MATERIALS AND METHODS: A questionnaire regarding the preferred investigation for acute flank pain was sent to all consultant urologists registered as British Association of Urological Surgeons members. RESULTS: Of the 548 consultants from 210 institutions surveyed, 293 (54%) from 171 (81.4%) institutions returned their questionnaires. Intravenous urography is used in the majority of institutions (146; 85.4%) for investigating acute flank pain. Only 18 (10.5%) use NESCT, while 4.1% use ultrasonography. Among those using IVU as the investigation of choice, the main reason given was limited CT services (82.4%). Others included familiarity with IVU features (51.2%), limited availability of radiologists for out-of-hours reporting of CT (26%), more rapid procedure (20.8%), lower cost (20%), and lower radiation exposure (19.6%). Only 52.4% of consultants using IVU would prefer NESCT if both were equally available. CONCLUSIONS: Urography remains the commonest modality for the investigation of acute flank pain in the UK and Ireland primarily because of limited CT services and greater familiarity with the images. Given the opportunity, only half of consultant urologists would select NESCT in preference to IVU, suggesting that improving the availability of CT services alone may not lead to practice paralleling that of our transatlantic and continental European colleagues. PMID- 16253059 TI - Impact of heads-up display imaging on endoscopic task performance. AB - BACKGROUND AND PURPOSE: Heads-Up Imaging goggles provide ergonomic advantages to the endourologist. This study was designed to evaluate whether heads-up display impacts task performance for ureteroscopic stone retrieval. MATERIALS AND METHODS: The ability to capture a 5-mm calculus with a Cook N-Circle 2.2F stone basket from an inanimate caliceal model was tested by three experienced and three novice stone-basket operators. Visual display for initial testing for each operator was randomized to the OptiVu HD3 Heads-Up googles or a 20- inch Sony Triniton monitor (TV). Subsequent testing alternated between the two devices. Camera input was provided by the Storz telecam SL-NTSC. The HD3 was set up to align the direction of view with the operator's hands, while the TV was aligned at an angle 45 degrees lateral and 30 degrees superior to the operator's direction of view to approximate the traditional room set-up for an endourologic procedure. Each operator performed five basketing trials with each display set up. RESULTS: Expert operators retrieved calculi more rapidly (9.2 +/- 5.9 seconds) than novice operators (50.7 +/- 48.9 seconds), irrespective of whether a TV monitor or goggle display was utilized as the imaging modality. No significant differences were noted in task performance between the two imaging modalities for the expert (P = b0.60), novice (P = 0.77), or overall (P = 0.91) groups. CONCLUSION: The Optiview Heads-Up goggle display system does not offer advantages in task performance with specific regard to the ability to capture stone fragments with baskets. PMID- 16253060 TI - Percutaneous nephrolithotomy for complete staghorn calculi in preschool children. AB - BACKGROUND AND PURPOSE: To evaluate the safety and efficacy of percutaneous nephrolithotomy (PCNL) for the management of complete staghorn calculi in children 45 degrees C in areas 3 to 4 mm from the antenna. However, the heat pattern and protocol of the TMx-2000 produce necrosis-generating conditions only within a few millimeters of the urethra. PMID- 16253075 TI - Effects of enalaprilat on the renin-angiotensin-aldosterone system and on renal function during CO2 pneumoperitoneum. AB - BACKGROUND AND PURPOSE: Mechanical and hormonal factors have been implicated in pneumoperitoneum-induced renal alterations. The aim of this study was to evaluate the effects of enalaprilat (Vasotec) administration on renal function during CO2 pneumoperitoneum, given that this drug, which is an angiotensin-converting enzyme inhibitor, by inhibiting the renin-angiotensin-aldosterone system, alters hormone induced changes during pneumoperitoneum. MATERIALS AND METHODS: Thirty adult dogs were randomly assigned to one of three groups (N = 10 each): group A (pneumoperitoneum not performed); group B (CO2 + enalaprilat); group C CO2 only. The groups were analyzed with consideration for body weight, hematologic values, hemodynamic parameters, and renal function (plasma renin activity, urinary debt, creatinine clearance, and sodium-excretory fraction). RESULTS: Hemodynamic and acid-basic parameter differences did not influence renal function. Plasma renin activity decreased significantly in group B compared with group C and stayed close to the values in group A. Creatinine clearance remained constant in group B, while in group C, creatinine clearance dropped, and this difference was statistically significant. Urinary debt and sodium-excretory fraction increased in group B during pneumoperitoneum and 60 minutes after this period in comparison with the other groups without reaching statistical significance. CONCLUSION: The decline in urinary debt and in creatinine clearance observed during pneumoperitoneum was less accentuated with administration of enalaprilat. PMID- 16253076 TI - Laparoscopic right nephrectomy using a novel technique to excise an inferior vena caval cuff in a porcine model. AB - BACKGROUND AND PURPOSE: Laparoscopic right nephrectomy has been performed using laparoscopic staplers to clamp and divide the renal vessels. In patients who have a tumor thrombus in the renal vein and in rightkidney donors, it is necessary to excise a cuff of the inferior vena cava (IVC) with the renal vein. We present a new technique for excising a cuff of the IVC and suturing it intracorporally using a new vascular clamp designed in our institution. MATERIALS AND METHODS: The vascular clamp was designed to be completely inserted into the peritoneal cavity through the hand-port incision. The renal vein with a cuff of the IVC was then excised, and the defect in the IVC was sutured with a vascular stitch intracorporally. This procedure was performed in the animal laboratory using a porcine model. RESULTS: A total of 20 hand-assisted right laparoscopic nephrectomies were performed. In the first five operations, the clamp dimensions and angles were modified until the ideal design was reached. In the next five operations, different suture materials and needle sizes were tried to find the best combination for the intracorporal IVC sutures. The first and second operations were not completed because of the difficulty in applying the initial version of the clamp to the IVC. In the 4th and 6th operations, bleeding occurred from the suture line: because of a missed stitch in the back wall of the IVC in one case and a needle stick to the IVC in the other case. The last 14 operations were successful without any bleeding or injuries. Suturing of the IVC was completed in 13 to 22 minutes. CONCLUSION: Right hand-assisted laparoscopic nephrectomy with excision of a cuff of the IVC using an intracorporal vascular clamp is safe and reproducible in a porcine model. In our hands, a learning curve of 10 cases was required. PMID- 16253077 TI - Multiple high-intensity focused ultrasound probes for kidney-tissue ablation. AB - BACKGROUND AND PURPOSE: To investigate kidney-tissue ablation by high-intensity focused ultrasound (HIFU) using multiple and single probes. MATERIALS AND METHODS: Ultrasound beams (1.75 MHz) produced by a piezoceramic element (focal distance 80 mm) were focused at the center of renal parenchyma. One of the three probes (mounted on a jig) could also be used for comparison with a single probe at comparable power ratings. Lesion dimensions were examined in perfused and unperfused ex vivo porcine kidneys at different power levels (40, 60, and 80 W) and treatment times (4, 6, and 8 seconds). RESULTS: At identical power levels, the lesions induced by multiple probes were larger than those induced by a single probe. Lesion size increased with increasing pulse duration and generator power. The sizes and shapes of the lesions were predictably repeatable in all samples. Lesions in perfused kidneys were smaller than those in unperfused kidneys. CONCLUSIONS: Ex vivo, kidney-tissue ablation by means of multiple HIFU probes offers significant advantages over single HIFU probes in respect of lesion size and formation. These advantages need to be confirmed by tests in vivo at higher energy levels. PMID- 16253078 TI - Effect of holmium:YAG laser pulse width on lithotripsy retropulsion in vitro. AB - BACKGROUND AND PURPOSE: The effect of laser pulse width on calculus retropulsion during ureteroscopic lithotripsy is poorly defined because of the limited availability of variable pulse-width lasers. We used an adjustable pulse-width Ho:YAG laser to test the effect of pulse width on in vitro phantom-stone retropulsion and fragmentation efficiency. METHODS AND MATERIALS: An Odyssey 30 Ho:YAG laser (Convergent Laser Technologies, Oakland, CA) with adjustable pulse width (350 or 700 microsec) was used to treat spherical 10-mm plaster calculi in a model ureter (N = 40) and calix (N = 16) utilizing 200- and 400-microm fibers (10 Hz, 1.0 J). Calculi were placed in a waterfilled clear polymer tube, and laser energy was applied continuously in near contact until the stone had moved 8 cm. The time (seconds) and energy (joules) needed to cause the stone to traverse this distance was recorded. Stones were also placed in a stainless-steel mesh calix model in which retropulsion was limited. Laser energy was applied for 5 minutes at each pulse width. A laser-energy meter (Molectron Detector Inc, Portland OR) was used to quantify fiber transmission efficiency after 1 minute of continuous lithotripsy for each fiber at each pulse width. RESULTS: Retropulsion was greater for stones treated at 350 microsec, indicated by a shorter time to traverse the model ureter. For the 200-microrm fiber at 350 microrsec, the average time was 11.5 seconds v 20.3 seconds at 700 microsec (P < 0.001). The average total energy delivered was 114.9 J at 350 microsec v 199.8 J at 700 microsec (P < 0.001). For the 400-microm fiber at 350 microsec, the average time was 5.8 seconds v 11.9 seconds at 700 microsec (P < 0.001). The average total energy was 57.1 J at 350 microsec v 127.3 J at 700 microsec (P < 0.001). In the caliceal model, at 350 and 700 microsec with the 200- and 400-microm fibers, mass loss was 34.9% and 33.4% (P = 0.8) and 14.6% and 21.6% (P = 0.04), respectively. The reduction in energy transmission at 350 microsec and 700 microsec with the 200- microm fiber after 60 seconds of continuous lasing was 8.82% v 9%, respectively (P = 0.95). For the 400-microm fiber, the transmission loss was 18.4% at 350 microsec v 4.4% at 700 microsec (P = 0.0002). CONCLUSION: When treating ureteral calculi, retropulsion can be reduced by using a longer pulse width without compromising fragmentation efficiency. For caliceal calculi, the longer pulse width in combination with a 400-microm fiber provides more effective stone fragmentation. PMID- 16253079 TI - LiteratureWatch. PMID- 16253082 TI - Evaluation of long-term implant success. AB - Implant success criteria, regarding marginal bone loss and other parameters, were first suggested in 1986 and today are still frequently referred to as the gold standard for implant success. However, according to the recent abundance of data on marginal bone loss and a better understanding of bone and soft tissue behavior around the implant neck and body, these criteria are inaccurate for the wide variety of implant systems. The purpose of this article is to review some of the relevant literature regarding marginal bone loss and to propose guidelines for a novel approach to evaluate the long-term success of implants regarding marginal bone loss. Four hypothetical patterns of implant marginal bone loss after the first year are suggested: a low-rate marginal bone loss over the years (Albrektsson's pattern); low-rate marginal bone loss in the first few years followed by a rapid loss of bone support; high-rate marginal bone loss in the first few years followed by almost no bone loss; and continuous high-rate marginal bone loss leading to a complete loss of bone support. PMID- 16253083 TI - Gingipain-specific IgG in the sera of patients with periodontal disease is necessary for opsonophagocytosis of Porphyromonas gingivalis. AB - BACKGROUND: Porphyromonas gingivalis is a primary etiologic agent of generalized aggressive periodontitis (GAgP), and gingipains, a group of cysteine proteinases, are critical virulence factors expressed by this organism. GAgP patients develop specific antibodies to gingipains; however, the function of these antibodies in the clearance of P. gingivalis infection is poorly understood. METHODS: In this study, we defined the levels of gingipain-specific antibodies in GAgP patient sera and examined the ability of gingipain-specific antibodies to facilitate opsonophagocytosis of P. gingivalis by human polymorphonuclear leukocytes (PMNs) using a fluorescent phagocytosis assay. RESULTS: GAgP patient sera possessed elevated levels of P. gingivalis-, arginine-gingipain (Rgp)A-, RgpB-, and lysine gingipain (Kgp)-specific IgG (Kgp > RgpA > P. gingivalis > RgpB). Adsorption of GAgP sera with P. gingivalis whole organisms, RgpA, RgpB, and Kgp conjugated to sepharose beads reduced opsonophagocytosis of P. gingivalis by PMNs. CONCLUSIONS: Our studies demonstrate that GAgP patient sera possess elevated levels of P. gingivalis- and gingipain-specific IgG. Furthermore, we show that gingipain antibodies promote uptake of P. gingivalis by PMNs, and our data suggest that gingipain-specific antibodies may be important for the control of P. gingivalis infections. PMID- 16253084 TI - Regenerative potential of platelet-rich plasma added to xenogenic bone grafts in peri-implant defects: a histomorphometric analysis in dogs. AB - BACKGROUND: The purpose of this investigation was to evaluate the regenerative influence of platelet-rich plasma (PRP) added to xenogenic bone grafts on bone histomorphometric parameters in a dog model. METHODS: Ninety endosseous dental implants were inserted in the mandibles of nine hound dogs. Mesial and distal 3 wall peri-implant defects were surgically created adjacent to the implants. Defects were randomly assigned to three groups: demineralized freeze dried bone with platelet-rich plasma (DFDB + PRP), DFDB alone, and no treatment (NT). Animals were sacrificed at 1, 2, and 3 months according to a previously established randomization schedule, and specimens were subjected to histomorphometric analysis. Percentages of bone area inside the implant threads (BiIT), bone-to-implant contact (BIC), and bone area outside implant threads (BoIT) were recorded. Treatment effects were evaluated using analysis of variance models. RESULTS: The effect of the three treatments on the outcome measures did not differ significantly by healing time (P > 0.05 for the healing time by treatment interaction). However, the average (standard deviation) percentage of BIC and BoIT was significantly different between the treatment groups. In particular, the average percentage of BIC differed between peri-implant defects treated with DFDB + PRP (33.8% [11.0]) and those treated with DFDB alone (28.5% [10.8]; P = 0.042), as well as those in the NT group (27.9% [11.0]; P = 0.024). Furthermore, the average percentage of BoIT differed significantly between defects treated with DFDB + PRP compared to defects in the NT group (51.6% [12.2] versus 43.3% [10.3]; P = 0.005). There was borderline evidence to suggest that the average percentage of BiIT and BIC was significantly different depending on the length of the healing time (P = 0.054 and P = 0.085, respectively). CONCLUSION: This study found that the addition of PRP to xenogenic bone grafts demonstrated a low regenerative potential in this animal model. PMID- 16253085 TI - Proteomic analysis of protein components in periodontal ligament fibroblasts. AB - BACKGROUND: Characterization of periodontal ligament (PDL) fibroblast proteome is an important tool for understanding PDL physiology and regulation and for identifying disease-related protein markers. PDL fibroblast protein expression has been studied using immunological methods, although limited to previously identified proteins for which specific antibodies are available. METHODS: We applied proteomic analysis coupled with mass spectrometry and database knowledge to human PDL fibroblasts. RESULTS: We detected 900 spots and identified 117 protein spots originating in 74 different genes. In addition to scaffold cytoskeletal proteins, e.g., actin, tubulin, and vimentin, we identified proteins implicated with cellular motility and membrane trafficking, chaparonine, stress and folding proteins, metabolic enzymes, proteins associated with detoxification and membrane activity, biodegradative metabolism, translation and transduction, extracellular proteins, and cell cycle regulation proteins. CONCLUSIONS: Most of these identified proteins are closely related to the extensive PDL fibroblasts' functions and homeostasis. Our PDL fibroblast proteome map can serve as a reference map for future clinical studies as well as basic research. PMID- 16253086 TI - The effects of three oral sprays on plaque and gingival inflammation. AB - BACKGROUND: The aim of this clinical trial was to compare the effects of three oral sprays containing chlorhexidine (CHX), benzydamine hydrochloride (B-HCl), and CHX plus B-HCl (CHX/B-HCl) on plaque and gingivitis. METHODS: Fifty-one periodontally healthy volunteers, randomly divided into three spray groups, refrained from all mechanical oral hygiene measures for 7 days and, instead, used one of the randomly assigned sprays twice daily. The plaque index (PI), gingival index (GI), and gingival bleeding time index (GBTI) were assessed at days 0 and 7. Side effects were also evaluated. RESULTS: In all groups, PI, GI, and GBTI showed significant increases from the baseline to day 7. There were no significant side effects for B-HCl spray and CHX spray at day 7, but, for CHX/B HCl, burning sensation significantly increased from the baseline. For all parameters, there were significant differences between B-HCl and CHX/B-HCl and between B-HCl and CHX, but no significant differences were observed between CHX/B HCl and CHX. In addition, a significant difference was found between CHX/B-HCl and CHX with regard to burning sensation. CONCLUSIONS: These results indicate that both CHX and CHX/B-HCl sprays have equal clinical effectiveness, but only B HCl spray has less anti-plaque and anti-gingivitis effects. Furthermore, CHX/B HCl spray causes more side effects. PMID- 16253087 TI - Involvement of Porphyromonas gingivalis fimA genotype in treatment outcome following non-surgical periodontal therapy. AB - BACKGROUND: Porphyromonas gingivalis is considered a critical pathogen in periodontal diseases. It is classified into six genotypes based on diversity of the fimA gene encoding fimbrillin. The present study evaluated the involvement of the fimA genotype in treatment outcome following non-surgical periodontal therapy. METHODS: Chronic periodontitis patients were enrolled in this study; all received clinical and microbiological examinations at baseline. The detection of subgingival species and identification of P. gingivalis fimA genotypes were performed using polymerase chain reaction based methods. In total, 160 P. gingivalis positive sites with bleeding on probing (BOP) and a probing depth of > or =4 mm were accepted. They were followed up after scaling and root planing. RESULTS: Longitudinal investigation indicated that fimA type I positive sites at baseline were followed by a significantly higher frequency of persistent BOP after treatment than type I negative sites (51.6% versus 27.9%), while types Ib and II were not. Type I positive sites also showed more persistence of Tannerella forsythensis and P. gingivalis after treatment than type I negative sites. In post-treatment investigation, type I positive sites showed higher frequencies of BOP and T. forsythensis detection than type I negative sites (77.8% versus 43.5% and 100% versus 76.1%, respectively). CONCLUSIONS: BOP in initially type I positive sites showed little improvement with treatment, and the combined persistence of fimA type I and T. forsythensis seemed to be involved in this poor treatment outcome. The present study demonstrated the potential of P. gingivalis fimA type I as a predictor of persistent BOP after treatment. PMID- 16253088 TI - Effect of recombinant human bone morphogenetic protein-2, -4, and -7 on bone formation in rat calvarial defects. AB - BACKGROUND: Currently, more than 20 bone morphogenetic proteins (BMPs) have been identified, and many trials have been carried out using recombinant human BMPs (rhBMPs) for bone tissue engineering. However, comparative analyses on bone formative activities of rhBMP using a preclinical model have been limited. Therefore, the aim of this study was to evaluate and compare the osteogenic potential of rhBMP-2, -4, and -7 delivered with absorbable collagen sponge (ACS) upon early (2 weeks) and complete (8 weeks) wound healing phases in a critical sized rat calvarial defect model. METHODS: Eight-millimeter critical sized calvarial defects were created in 30 male Sprague-Dawley rats. The animals were divided into three groups of 10 animals each. The defects were treated with 0.025 mg/ml rhBMP-2/ACS, rhBMP-4/ACS, or rhBMP-7/ACS. The rats were sacrificed at either 2 (five rats) or 8 (five rats) weeks after surgery, and the results were evaluated histologically, histomorphometrically, and immunohistometrically. RESULTS: The surgical implantation of rhBMP-2/ACS, rhBMP-4/ACS, or rhBMP-7/ACS resulted in enhanced local bone formation in the rat calvarial defect model at both 2 and 8 weeks. The amount of defect closure, new bone area, and bone density were similar in the three groups at each time point (P > 0.05). In terms of bone density and new bone area, there were statistically significant differences between results obtained at 2 weeks and those obtained at 8 weeks in all groups (P < 0.05). Two-way analysis of variance (ANOVA) revealed that there was no correlation between the time and conditions (P > 0.05), but time was found to have a strong influence on defect closure, new bone area, and bone density (P < 0.05). Irrespective of rhBMP type, positive immunoreactions of osteopontin (OPN) and osteocalcin (OCN) were evident at 2 and 8 weeks. Intense OPN and OCN staining was observed near the newly formed bone as well as in some cells within the new bone. CONCLUSIONS: Within the rhBMP types used, rhBMP concentration, and the observation interval, there appears to be no specific differences in bone regenerative potential. All rhBMPs used in this study may be considered effective factors for inducing bone formation. PMID- 16253089 TI - T cells support osteoclastogenesis in an in vitro model derived from human periodontitis patients. AB - BACKGROUND: Periodontitis is characterized by alveolar bone destruction; however, the mechanisms responsible for bone damage are poorly understood. It has been reported that T cells are implicated in the pathogenesis of periodontitis. It has been also demonstrated that activated T lymphocytes secrete receptor activator of nuclear factor-kappa B ligand (RANKL) and can support the differentiation of monocytes into resorbing osteoclasts (OCs). Therefore, the purpose of this study was to examine the OC formation in periodontitis patients (PP) and the role of T cells in osteoclastogenesis. METHODS: To study OC formation, we used an in vitro model consisting of unstimulated and unfractionated peripheral blood mononuclear cells (PBMCs) from PP and controls. In parallel, T-cell-depleted PBMCs from the same patients were also established. The expression of RANKL and tumor necrosis factor-alpha (TNF-alpha) was analyzed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot in fresh T cells isolated from PP and controls. Functional antibodies, anti-RANKL and anti-TNF-alpha, were utilized to study osteoclastogenesis in PBMC cultures from PP. RESULTS: We showed that, in unfractionated PBMCs from PP, the OCs spontaneously developed in a T-cell dependent way. The addition of macrophage colony stimulating factor (MCSF) and RANKL was necessary to promote the osteoclastogenesis in T-cell-depleted PBMC cultures from PP and in unfractionated PBMCs from periodontally healthy controls. Moreover, freshly isolated T cells from PBMCs of PP overexpressed RANKL and TNF alpha. Finally, functional anti-RANKL and anti-TNF-alpha antibodies significantly inhibited osteoclastogenesis. CONCLUSION: Our data suggest that T cells support spontaneous osteoclastogenesis in PP via RANKL and TNF-alpha overexpression. PMID- 16253090 TI - Repair of Class II furcation defects after a reparative tissue graft obtained from extraction sockets treated with growth factors: a histologic and histometric study in dogs. AB - BACKGROUND: Reparative tissue of extraction sockets was proposed as grafting material in the treatment of periodontal defects. Our hypothesis was that the addition of growth factors to extraction sockets improves the regenerative potential of this tissue when used as a graft. The objective of the present study was to analyze qualitatively and quantitatively the repair of acute Class II furcation defects after they receive this grafting material. METHODS: The second and third upper premolars were extracted from four dogs. Platelet-derived growth factor (PDGF)-BB and insulin-like growth factor (IGF)-I, at concentrations of 6 microg/ml each, were applied to the resulting sockets. After 5 days, 24 acute defects (12 control and 12 test defects) were created in the second, third, and fourth lower premolars. Only the test sites received the graft. The flaps were positioned coronally on both sides and sutured. After 45 days, the specimens were collected, decalcified, and processed histologically in a buccal-lingual plane. The parameters were measured horizontally in the buccal-lingual direction. RESULTS: Repair was histologically and histometrically similar in the two groups. No significant difference was observed between the test and control groups in the parameters connective tissue, new cementum, new bone, and junctional epithelium. CONCLUSION: The use of this graft did not show beneficial effects on the repair of acute Class II furcation defects in dogs. PMID- 16253091 TI - Periodontal regeneration with peptide-enhanced anorganic bone matrix in particulate and putty form in dogs. AB - BACKGROUND: Peptide-enhanced (using the peptide 15 [P-15] synthetic peptide) anorganic bone matrix (ABM) particulate (PPart) grafts have demonstrated clinical and histologic success in human periodontal defects. Dispersion of ABM/P-15 in sodium hyaluronate carrier (PPutty) improves the handling properties of the graft material. The healing of ABM/P-15 particulate and ABM/P-15 putty was compared in critical-sized fenestration defects in 16 mongrel dogs. METHODS: After full thickness flap reflection, 7 mm diameter fenestrations were made with a trephine in the mid-root of both maxillary canines in each dog. Bone, periodontal ligament, and cementum were removed as completely as possible with hand root planing within the trephine-produced notches. By random allocation, each defect was filled with PPart or PPutty, and the flaps were closed with sutures. One dog contributing two defects served as a negative control. Block sections were retrieved at 3 and 8 weeks for histologic processing. Three 6-mu step serial sections in the center of the defects were used for analysis. Parameters measured included the original length of the wound, linear amount of periodontal regeneration including new cementum, bone and connective tissue, and area measurements of new bone and remaining particles. PPutty and PPart results were compared for significant differences using the Wilcoxon rank sum test. RESULTS: Clinical healing was uneventful in all cases. There was no evidence of inflammation or adverse tissue reactions with either material. The controls showed minimal regeneration at the periphery of the defect. Histomorphometric evaluation of the grafted defects revealed the following: at 3 weeks, there was minimal new bone formation (occupying 4.2% of the grafted area for the PPutty and 1.2% for the PPart). The grafted particles occupied 21.2% and 35.6% of the area for the PPutty and PPart, respectively (P = 0.039). At 8 weeks, there was a tendency for greater new bone formation compared to 3 weeks with both materials. There was significantly more new bone with the PPutty (49.3%) compared to the PPart (14.8%) (P = 0.045). The grafted particles occupied 7.9% and 17% of the grafted area for the PPutty and PPart, respectively (no significant difference). There were no significant differences for any of the linear measurements. ABM/P 15 PPutty had superior handling characteristics. CONCLUSIONS: Both ABM/P-15 materials yielded satisfactory healing and resulted in the greater regeneration of fenestration defects in dogs at 8 weeks compared to controls. In addition, AMB/P-15 putty resulted in more bone formation compared to ABM/P-15 particulate. PMID- 16253093 TI - Biological factors involved in the osseointegration of oral titanium implants with different surfaces: a pilot study in minipigs. AB - BACKGROUND: The stability of titanium implants is determined by the rigid load bearing connections that are formed by the bone, a process that involves a complex network of cells, pro- and anti-inflammatory mediators, and growth factors. The osseointegration processes at the interfaces of machined and porous implants were studied using molecular and histological techniques. METHODS: Two machined and two porous titanium implants were inserted into the tibiae of four minipigs. The animals were sacrificed at 15, 30, 60, and 90 days post implantation. The levels of bone morphogenetic protein (BMP)-4, transforming growth factor (TGF)-beta1, and tumor necrosis factor (TNF)-alpha were quantified in the peri-implant osseous samples. The levels of interleukin (IL)-1beta, IL-6, IL-10, and TNF-alpha in the serum were also assessed. RESULTS: Histomorphological analysis showed evidence of bone ossification around the porous implant at 60 days. Surrounding the machined implants, highly sclerotic fibrous pads started the healing response at 90 days, and the levels of TGF-beta1 and BMP-4 began to increase at 60 days, at which time bone ossification around the porous implants was already evident. TNF-alpha was not present in the bone next to the implants. The serum levels of cytokines IL-1beta, IL-6, and IL-10 were not increased. The serum level of TNF-alpha increased during the healing process. CONCLUSIONS: We observed that the levels of BMP-4 and TGF-beta1, which play essential roles in the osteogenesis process, increased earlier around the porous implants than around the machined implants. Similarly, the ossification process was initiated earlier at the surfaces of the porous implants than at the surfaces of the machined implants. PMID- 16253092 TI - The effect of sputtered calcium phosphate coatings of different crystallinity on osteoblast differentiation. AB - BACKGROUND: Coating titanium implants with hydroxyapatite (HA) has been suggested to increase osseointegration by stimulating early osteoblast function. The goal of this study was to determine the extent to which the crystalline content of the HA surface affected osteoblast function in vitro. METHODS: Osteoblasts were isolated from fetal rat calvaria. Titanium coupons were sputter coated and analyzed. Mineralized nodule formation on plastic using von Kossa staining was compared to tetracycline and procion dye labeling. Cell proliferation, adhesion, alkaline phosphatase activity, morphology and spreading, and cytoskeletal arrangement were analyzed. Reverse transcription-polymerase chain reaction (RT PCR) was used to determine the expression of mRNA for specific proteins. RESULTS: The percent crystallinity of coatings was 0% (HA1), 1.9% +/- 0.4% (HA2), and 66.4% +/- 2.8% (HA3). The nodule formation and cell number were greatest on titanium and HA3 compared to HA1 and HA2 (P < 0.01). At weeks 2 to 4, all samples showed strong alkaline phosphatase, osteocalcin, monocyte-colony stimulating factor (M-CSF), and receptor activator of nuclear factor kappa B ligand (RANKL) expression, but the specific activity of alkaline phosphatase decreased. Cell adherence was greater than 60% of applied cells for all surfaces except HA3. The cells were significantly more elongated on titanium, with no difference on the HA coated surfaces. Actin filaments were arranged peripherally at 5 hours but arranged parallel to the long axis of the cell at 20 hours. CONCLUSIONS: Procion labeling is a valid method for evaluating mineralized nodule formation on opaque surfaces. There were no major differences in osteoblast function using titanium or high-crystalline coatings, and most functions were decreased on amorphous or low-crystalline coatings. PMID- 16253094 TI - Overweight and obesity as risk indicators for periodontitis in adults. AB - BACKGROUND: Developed and developing countries are facing an obesity epidemic with various health consequences. Few studies have addressed the relationship between obesity and periodontal health. The present study assessed the association of overweight and obesity with periodontitis in Brazilian adults. METHODS: A representative probability sample comprising 706 subjects aged 30 to 65 years from south Brazil was examined clinically and using a structured interview. Overweight and obesity were assessed by body mass index (BMI) using the World Health Organization (WHO) criteria. Individuals with > or =30% teeth with attachment loss > or =5 mm were classified as having periodontitis. Statistical analysis accounted for survey design, and separate analyses were performed for non-smokers. RESULTS: In this population, 60% and 65% of males and females, respectively, were overweight or obese. Periodontitis was observed in 50.7% and 35.3% of males and females, respectively. The percentage of males with periodontitis was similar in the overweight/obese individuals compared to those with normal weight. In females, there was a positive correlation between the BMI index and the occurrence of periodontitis, with a significantly (P < 0.05) higher prevalence of periodontitis in obese than in normal weight females. The multivariable analysis showed that obese females were significantly more likely (odds ratio = 2.1) to have periodontitis than normal weight females. A separate analysis for non-smokers showed that obese females were approximately 3.4 times more likely to have periodontitis than the normal BMI group. There were no significant differences in the prevalence of periodontitis between BMI groups among smokers of both genders and in male non-smokers. CONCLUSIONS: Obesity was significantly associated with periodontitis in adult, non-smoker women. Overweight was not significantly associated with periodontitis. Smoking may attenuate the association of periodontitis with obesity. PMID- 16253095 TI - Factors affecting the outcomes of coronally advanced flap root coverage procedure. AB - BACKGROUND: The coronally advanced flap (CAF) has been used to treat gingival recession. However, the final outcomes (percentage of root coverage) vary from case to case. Hence, the purpose of this study was to analyze the factors that may affect the results of CAF root coverage procedures. METHODS: Twenty-three systemically healthy patients (mean age, 43.8 +/- 11.9 years) each with one Miller's Class I buccal recession defect were included. Baseline clinical parameters included recession depth (RD), recession width (RW), gingival thickness (GT), width of keratinized tissue (WKT), clinical attachment level (CAL), probing depth (PD), plaque index (PI), and gingival index (GI). CAF root coverage procedures were performed to correct the recession defects. Patients were followed at 2, 4, 12, and 24 weeks post-surgery, at which time wound healing index (WHI) and other measurements were recorded. RESULTS: The mean baseline RD was 2.9 +/- 0.4 mm; RW, 3.4 +/- 0.6 mm; WKT, 2.7 +/- 1.3 mm; and GT, 1.1 +/- 0.3 mm. At mid-buccal, the mean CAL was 4.5 +/- 0.8 mm. Six months after surgery, the average RC was 82.3% +/- 24.7%; RD, 0.5 +/- 0.7 mm; RW, 0.4 +/- 0.9 mm; WKT, 3.2 +/- 0.9 mm; and GT, 1.5 +/- 0.5 mm. At mid-buccal, the mean CAL was 1.8 +/- 1.1 mm. From baseline to the 6-month follow-up, the changes of RC, RD, RW, WKT, GT, and CAL showed statistical significance (P < 0.05). Fourteen patients achieved 100% RC. The mean RC in partial coverage cases was 54.8% +/- 16.8%. Analysis revealed that an initial GT thicker than 1.2 +/- 0.3 mm was associated with complete root coverage at the 6-month follow-up (P < 0.05). CONCLUSIONS: CAF is a predictable procedure to treat Miller's Class I mucogingival defects. Initial GT was the most significant factor associated with complete root coverage. PMID- 16253097 TI - Human gingival fibroblast integrin subunit expression on titanium implant surfaces. AB - BACKGROUND: Implant surface characteristics have been shown to modify cell behavior and regulate integrin expression. Integrin expression and resultant integrin-mediated cellular activity are essential components of tissue healing and homeostasis. Although both osseous and soft tissue healing around dental implants are critical to clinical success, there is limited information available on the effect of implant surfaces on integrin expression in soft tissues. Therefore, the aim of this study was to examine integrin expression for gingival fibroblasts on titanium surfaces and the influence of titanium surface roughness on integrin expression and cell morphology. METHODS: Human gingival fibroblasts were cultured on smooth (polished) and rough (sand-blasted acid-etched) titanium surfaces and a cell culture plastic (control) surface. To analyze integrin expression, total RNA was isolated from experimental and control cells, and levels of integrin subunit mRNA were assessed by reverse transcription-polymerase chain reaction (RT-PCR) using primers specific for the alpha2, alpha4, alpha5, alpha(v), and beta1 integrin subunits and aldolase (internal control). PCR products were analyzed by polyacrylamide gel electrophoresis (PAGE), confirmed via DNA sequencing, and quantified using computer-assisted densitometry. The expression of the integrin subunits was analyzed at the protein level using flow cytometry, as well as fluorescence and confocal laser microscopy. Cell morphology was evaluated using scanning electron microscopy (SEM). RESULTS: Our experiments demonstrated cellular expression of the alpha2, alpha4, alpha5, alpha(v), and beta1 integrin subunits at both mRNA and protein levels on all surfaces. In addition, the alpha4 and beta1 mRNA levels were significantly increased on smooth titanium relative to plastic surfaces (P <.05) with intermediate mRNA levels found on the rough titanium surfaces. The smooth titanium surfaces exhibited a flat monolayer of cells, while rough titanium surfaces showed cells orienting themselves along surface irregularities. CONCLUSIONS: These results demonstrate the presence of multiple integrin subunits in human gingival fibroblasts grown in contact with titanium implant surfaces and that titanium surface roughness alters cellular morphology but appears to have limited effects on integrin expression. This study provides insight into the complicated cellular and molecular events occurring at the implant surface that may be critical to optimizing the soft tissue interactions with the soft tissue-implant interface. PMID- 16253096 TI - Prostaglandin production by human gingival fibroblasts inhibited by triclosan in the presence of cetylpyridinium chloride. AB - BACKGROUND: The effect of triclosan plus the cationic detergent cetylpyridinium chloride (CPC) was evaluated for prostaglandin inhibition in human gingival fibroblasts. Since triclosan has previously been shown to inhibit proinflammatory cytokine induced prostaglandin E2 (PGE2) production, we wanted to determine if triclosan, in the presence of CPC, could enhance these effects. METHODS: Initial studies determined that both triclosan and CPC were cytotoxic to human gingival fibroblasts in concentrations exceeding 1.0 microg/ml for either agent longer than 24 hours in a tissue culture. Therefore, subsequent studies measuring prostaglandin biosynthesis and cyclooxygenase (COX)-1 and COX-2 mRNA expression were performed in concentrations and times that did not significantly affect cell viability. RESULTS: PGE2 biosynthesis was dose dependently inhibited by both triclosan and triclosan and CPC when challenged by tumor necrosis factor (TNF) alpha or interleukin (IL)-1beta. At pharmacologically relevant concentrations, triclosan and CPC inhibited IL-1beta-induced PGE2 production to a greater extent than triclosan alone (P = 0.02). Moreover, enhanced COX-2 mRNA repression was observed with triclosan and CPC in comparison to triclosan alone in IL-1beta and TNF-alpha stimulated cells. No effect on COX-1 gene expression was observed. Further analysis of cell signaling mechanisms of triclosan and CPC indicates that nuclear factor-kappa B (NF-kappaB) and not p38 mitogen-activated protein kinase (MAPK) signaling may be impaired in the presence of triclosan and CPC. CONCLUSION: This study indicates that triclosan and CPC are more effective at inhibiting PGE2 at the level of COX-2 gene regulation, and this combination may offer a potentially better anti-inflammatory agent in the treatment of inflammatory lesions in the oral cavity. PMID- 16253098 TI - Oral health impacts on daily living related to four different treatment protocols for chronic periodontitis. AB - BACKGROUND: The aims of this study were to evaluate the oral health impacts perceived by patients submitted to different treatments of chronic periodontitis and their association with clinical parameters. METHODS: Sixty patients were assigned to one of the following therapeutic groups: control, treated with full mouth scaling and root planing (SRP); test 1, treated with SRP and 400 mg systemically administered metronidazole (MET) three times per day for 10 days; test 2, treated with SRP and professional supragingival plaque removal (PP) every week for 3 months; and test 3, treated with SRP and MET plus PP. Clinical periodontal measurements and data regarding patients' oral health impacts (perceived impacts on bleeding gums, gingival recession, sensitivity to cold, packing foods, aesthetics, bad breath, and tooth mobility) were collected at baseline and 3 months after therapy. RESULTS: All groups presented significant improvement in oral health perceived impacts. There was no statistically significant difference in the improvement of oral health impacts among groups subjected to different treatments. The clinical data of percentage of deep probing depth, deep clinical attachment level, and bleeding on probing were found to be correlated significantly with oral health impacts. CONCLUSIONS: Periodontal treatment leads to a significant reduction of self-perceived impacts regardless of the non-surgical treatment protocol employed. Most of the clinical data were associated with oral health impacts. PMID- 16253099 TI - A comparison of three techniques to obtain root coverage on mandibular incisors. AB - BACKGROUND: Multiple variations on the subepithelial connective tissue graft have been shown to produce good clinical results. The goal of this study was to compare three variations in the treatment of recession on mandibular incisors. METHODS: An a priori analysis was done to determine the sample size. Three groups of patients received root coverage procedures on mandibular incisors. Each group was treated with a different variation of a subepithelial graft. The first group received a connective tissue (CT) graft with a coronally positioned flap (CPF) (CPF + CT). The second group received a connective tissue graft with a double pedicle (DP) graft (DP + CT). The third group received a connective tissue graft with a tunneling (TUN) procedure and a laterally positioned (LAT) pedicle (TUN LAT + CT). The clinical results of these procedures were compared to determine if one procedure offered an advantage over the other procedures. RESULTS: All of the procedures produced a statistically significant improvement in the clinical parameters. The groups treated with the DP + CT and TUN-LAT + CT had greater mean root coverage (95.5% and 90.5%, respectively) than the CPF + CT group (80.2%). Additionally, DP + CT produced a greater increase in keratinized tissue (3 mm) than CPF + CT (1.4 mm) or TUN-LAT + CT (1.9 mm). There was no statistically significant difference in the mean root coverage of any technique treating defects <3 mm deep (CPF + CT, 90.9%; DP + CT, 96.4%; and TUN-LAT + CT, 92.1%) or the defects >3 mm deep treated with DP + CT (95.4%) or TUN-LAT + CT (88.3%). However, the CPF + CT produced less mean root coverage (68.4%) when treating defects > or =3 mm deep. When defects treated as isolated defects were compared to cases where multiple defects were treated, the cases with multiple defects treated with the CPF + CT had less mean root coverage (77%) than cases where single defects were treated with the CPF + CT (90.3%), DP + CT (isolated, 96.7% and multiple, 95.6%), and TUN-LAT + CT (isolated, 97.2% and multiple, 87.8%). CONCLUSIONS: All three of the procedures were effective in obtaining root coverage and improved clinical parameters on mandibular incisors. Overall, the DP + CT and TUN-LAT + CT procedures had greater mean root coverage than the CPF + CT technique. Based on this study, when treating defects > or =3 mm deep, one should consider using the DP + CT or TUN-LAT + CT rather than the CPF + CT. Additionally, when treating multiple defects at a time, one should consider using the DP + CT or TUN-LAT + CT rather than the CPF + CT. In cases where an increased amount of keratinized tissue is desired, based on this study, the DP + CT may be the best procedure to use. PMID- 16253101 TI - Biological evaluation of chitosan nanofiber membrane for guided bone regeneration. AB - BACKGROUND: Chitosan is known as a biodegradable and non-toxic natural polymer that enhances wound healing and bone formation. The aims of this study are to evaluate the biocompatibility of chitosan nanofiber membranes and to examine the effect of the chitosan nanofiber membranes on bone regeneration in rabbit calvarial defects. METHODS: In vitro cell proliferation tests using human osteosarcoma cell line MG63 and reverse transcription-polymerase chain reaction (RT-PCR) to evaluate the expression of alkaline phosphatase (ALP), collagen, osteocalcin (OCN), and GAPDH were done on chitosan nanofiber membranes. Chitosan nanofiber membranes were implanted in subcutaneous connective tissue and observed for 2, 4, and 6 weeks in rats. Ten-millimeter diameter round cranial defects were made in rabbits and covered by chitosan nanofiber membranes for 4 weeks. Specimens were decalcified and observed by a light microscope. RESULTS: MG63 cells proliferated for 28 days on the chitosan nanofiber membranes and expressed ALP, collagen, OCN, and GAPDH at 2 weeks. Chitosan nanofiber membranes that were grafted into rat subcutaneous tissue maintained their shape and space for bone regeneration for as long as 6 weeks. No inflammation could be seen on the membrane surface or in the surrounding tissues. In the rabbit calvarial defects, new bone filled the defects and fused to the original old bone in 4 weeks. CONCLUSIONS: The biocompatibility of the chitosan nanofiber membrane was confirmed, with enhanced bone regeneration and no evidence of an inflammatory reaction. This experiment shows that the novel biodegradable chitosan nanofiber membrane may be useful as a tool for guided bone regeneration. PMID- 16253100 TI - The effect of platelet-rich plasma on the coronally advanced flap root coverage procedure: a pilot human trial. AB - BACKGROUND: Coronally advanced flap (CAF) has been shown to effectively treat gingival recession. Platelet-rich plasma (PRP), containing autologous growth factors, has been shown to promote soft tissue healing. Therefore, the purpose of this study was to evaluate the effects of PRP in combination with CAF. METHODS: Twenty-four systemically healthy patients participated in this study. A single Miller's Class I buccal recession defect per patient was treated. These patients were randomly assigned into CAF or PRP + CAF groups. Clinical parameters included recession depth (RD), recession width (RW), gingival thickness (GT), width of keratinized tissue (WKT), clinical attachment level (CAL), probing depth (PD), plaque index (PI), wound healing index (WHI), and gingival index (GI). PRP was prepared from whole blood drawn prior to surgery and applied to root surfaces. Patients were followed at 2, 4, 12, and 24 weeks post-surgery. RESULTS: Twenty three patients completed the study. The RD at 24 weeks was significantly reduced from 2.9 +/- 0.5 to 0.5 +/- 0.6 mm in the CAF group (P < 0.05) and from 2.8 +/- 0.2 to 0.5 +/- 0.7 mm in the PRP + CAF group (P < 0.05). The mean root coverage was 83.5% +/- 21.8% in the CAF group and 81.0% +/- 28.7% in the CAF + PRP group (P > 0.05). Fourteen out of 23 patients (60.9%) experienced 100% root coverage at the 24-week postoperative follow-up. CONCLUSION: Based on the results of this pilot study, the application of PRP in CAF root coverage procedure provides no clinically measurable enhancements on the final therapeutic outcomes of CAF in Miller's Class I recession defects. PMID- 16253102 TI - A comparison between enamel matrix derivative and a bioabsorbable membrane to enhance healing around transmucosal immediate post-extraction implants. AB - BACKGROUND: This clinical report compares the use of an enamel matrix derivative (EMD) and bioabsorbable barrier membrane to enhance healing following the immediate placement of transmucosal implants into extraction sockets. METHODS: Thirty-two adult patients scheduled for tooth replacement with dental implants agreed to participate. Following the insertion of a transmucosal implant into the extraction site, the subjects were assigned to one of two treatment alternatives of the remaining bone defects around the implants: 1) the residual bone defects were filled with EMD (EMD group) or 2) the residual bone defects were covered with a bioabsorbable membrane (membrane group). Flaps were then coronally positioned around implant cover screws. Patients followed weekly maintenance recalls for the first 6 weeks and then monthly recalls until the final prosthetic restoration was completed (after 6 months). The treatment outcome was evaluated after 12 months by the use of clinical variables. The null hypothesis of no treatment group differences was tested by the use of analysis of variance (ANOVA). RESULTS: At a 12-month follow-up, all of the implants were completely osseointegrated and successfully functioning, showing a success rate of 100%. The membrane group showed a significantly lower mean probing attachment level than the EMD group at proximal (0.60 mm, standard deviation (SD) 0.37 versus 1.19 mm, SD 1.10), buccal (0.80 mm, SD 0.79 versus 1.77 mm, SD 1.16), and lingual sites (0.44 mm, SD 0.52 versus 1.48 mm, SD 1.46). The difference was statistically significant at all sites (P < 0.05). With respect to the position of the soft tissue margin around the implant shoulder, the membrane group showed a consistently higher value than the EMD group at, respectively, proximal (1.30 mm, SD 2.37 versus 1.16 mm, SD 1.0), buccal (0.90 mm, SD 1.29 versus 0.22 mm, SD 1.47), and lingual sites (1.12 mm, SD 1.10 versus 0.55 mm, SD 1.42). CONCLUSIONS: The membrane group obtained more favorable results in terms of both the probing attachment level and peri-implant position of soft tissues compared to the EMD group. The use of a bioabsorbable membrane around immediately placed transmucosal implants enhanced soft and hard tissue healing and might be an advisable treatment choice particularly in areas with high esthetic demands. PMID- 16253103 TI - Iatrogenic trauma to oral tissues. AB - BACKGROUND: Iatrogenic trauma can be defined as any trauma that has been induced by the dentist's activity, manner, or therapy. The aim of this article is to present traumatic oral tissue lesions of iatrogenic origin. METHODS: Thirteen cases of chemical (due to ferric sulfate and formocresol), physical (due to orthodontic wires and appliances), and thermal (due to electrosurgery) injuries to the oral tissues are reported. RESULTS: Chemical, physical, and thermal injuries in the oral, gingival, or palatinal mucosa of iatrogenic origin can exhibit various clinical features. The management of traumatic injuries is dependent on the severity of the involvement in the periodontal tissues. While, in most cases, the elimination of the offending agent and symptomatic therapy were sufficient, in severe cases, or when the injury resulted in permanent defects, periodontal surgery and regenerative therapy may be necessary. CONCLUSIONS: The skill, experience, and up-to-date knowledge of dentists are the main factors to prevent possible iatrogenic traumas. Although "To err is human," careful practice is very important for the principle "Primum non nocere" ("First do no harm"). PMID- 16253104 TI - Squamous cell carcinoma presenting as an endodontic-periodontic lesion. AB - BACKGROUND: Regardless of advances in diagnosis and treatment during the past 40 years, the overall 5-year survival rates for oral and oropharyngeal squamous cancers have only slightly improved and remain around 50%. Thus, the early diagnosis and treatment of carcinoma by health care providers are essential in achieving a good prognosis. We report a case of invasive squamous cell carcinoma that presented as a benign endodontic-periodontic lesion with a 7-mm periodontal pocket on tooth #15 in a 40-year-old, non-smoking woman. The subsequent management of the case is also discussed. The study was conducted in accordance with the Helsinki Declaration of 1975, as revised in 2000. METHODS: Our patient was seen for a comprehensive periodontal examination including a periodontal charting, occlusal analysis, study casts, electronic pulp test for tooth #15, and complete mouth periapical radiographs. As there was a periapical radiolucency, an endodontic consultation was obtained. A periodontal flap surgical procedure was performed on teeth #13 to #15, and as there was bone erosion into the maxillary sinus, a biopsy of the soft tissue was submitted to the local hospital for histological analysis. RESULTS: The biopsied lesion was diagnosed as invasive, moderately differentiated squamous cell carcinoma with focal spindle and clear cell differentiation (grade II to III of IV). Bone invasion was also identified. The treatment of the carcinoma involved a hemimaxillectomy with the removal of the maxillary left posterior teeth. The patient remained free of tumor for 5 years after the initial presentation. CONCLUSIONS: Patient education and periodic oral cancer examinations by dental professionals are necessary to reduce diagnostic delay and improve prognosis. This case report emphasizes the important role of dental professionals, especially periodontists and endodontists, of being aware that squamous cell carcinoma may manifest itself clinically and/or radiographically as a common periodontal or endodontic lesion. PMID- 16253105 TI - Constituting community: creating a place for oneself. PMID- 16253106 TI - The healing power of relationships. PMID- 16253107 TI - The value of social contacts. PMID- 16253108 TI - Community re-entry: development of life skills. PMID- 16253109 TI - Weak ties and schizophrenia: promise and problems. PMID- 16253110 TI - "If you want to bring someone back to reality, you have to enter their world first". PMID- 16253111 TI - More fundamentally human than otherwise. PMID- 16253113 TI - Changes in representations of a self-designated significant other in long-term intensive inpatient treatment of seriously disturbed adolescents and young adults. AB - Blatt and colleagues (1996) found that severity of psychopathology in seriously disturbed, treatment-resistant, hospitalized adolescents at the beginning of treatment was positively correlated with the degree to which these adolescents were involved in describing their parents. At the end of long-term, intensive, psychodynamically oriented, inpatient treatment of these very troubled adolescents, reduction in the severity of psychopathology correlated significantly with increases in the development of the structural organization of descriptions of mother, father, self, and therapist. These findings suggested that treatment of seriously disturbed, treatment-resistant, adolescent and young adult inpatients seems to involve at least two primary dimensions: 1) disengagement from an intense involvement with parents and 2) development in the structural organization of representations of self and a significant new figure, the therapist. The present study extends these earlier findings by examining changes in the description of a "significant other" that each patient elected to describe at the beginning and the end of treatment. Clinical improvement over the course of treatment was significantly correlated with developmental progression of the significant figure each patient selected to describe (from a grandparent to a close friend) as well as with progression in the developmental organization in which this significant other was described. These findings suggest that treatment of seriously disturbed adolescents and young adults involves a disengagement from an intense involvement with primary caregivers to involvement with others outside the family matrix and the developmental elaboration of the representation of these figures. PMID- 16253112 TI - Differences in mental health outcomes among Whites, African Americans, and Hispanics following a community disaster. AB - A number of studies have assessed the association between race and ethnicity and psychological health status following exposure to a stressful event. However, some of these studies indicate racial and ethnic minorities have poorer mental health relative to Whites, while others show no differences or that minorities may actually have better psychological health. One year after the terrorist attacks on the World Trade Center, we collected data on a random sample of city residents (N = 2368). The dependent variables were posttraumatic stress disorder (PTSD), PTSD symptom severity, major depression, panic attack, and general physical and mental well-being. We categorized our respondents as Non-Hispanic White, Non-Hispanic African American, Dominican, Puerto Rican, and Other Hispanics. Bivariate results indicated racial/ethnic differences for PTSD symptom severity, depression, general physical and mental health, and panic attack. Using logistic regression and controlling for possible confounding factors, most of these associations were rendered non-significant. That is, we found no post disaster racial/ethnic differences for PTSD, PTSD symptom severity, or physical health. African Americans and Other Hispanics were less likely to meet criteria for major depression or to be classified as unhealthy on the self-report SF-12 mental health scale compared to Whites. Only for panic attack were African Americans and Puerto Ricans more likely to meet criteria for this outcome. Thus, our study found little support for the hypothesis that Latinos or African Americans consistently suffered from poorer psychological and physical well-being in the aftermath of traumatic events, relative to Whites. PMID- 16253114 TI - War correspondents as responders: considerations for training and clinical services. PMID- 16253117 TI - Generation and characterization of a high-affinity chimaeric antibody against hepatitis B surface antigen. AB - Antibody against HBsAg (hepatitis B surface antigen) is advocated for the passive immunotherapy in certain cases of hepatitis B infections. A recombinant monoclonal antibody against HBsAg would offer several advantages over the currently used polyclonal human hepatitis B immunoglobulin. 5S is a mouse monoclonal antibody that binds to HBsAg with very high affinity. However, this mouse antibody cannot be used for therapeutic purposes, as it may elicit antimouse immune responses. Chimaerization, by replacing mouse constant domains with human counterparts, can reduce the immunogenicity of this molecule. We have cloned the V(H) (heavy-chain variable region) and V(L) (light-chain variable region) genes of this mouse antibody, and fused them with C(H)1 (heavy-chain constant domain 1) of human IgG1 and C(L) (light-chain constant domain) of human kappa chain respectively. These chimaeric genes were cloned into a mammalian expression vector (pFab-CMV), which has a modular cassette coding for part of the hinge, C(H)2 and C(H)3 of human IgG1. The recombinant construct was transfected in CHO (Chinese-hamster ovary) cells to generate a stable transfectoma. The resulting transfectoma was maintained in a serum-free medium and the full-length chimaeric anti-HBsAg antibody was purified from the culture supernatant. The yield of the purified chimaeric antibody was moderate ( approximately 5.5 mg/l). We further characterized the chimaeric antibody using several in vitro techniques. It was observed that the chimaeric molecule was glycosylated and expressed in the expected heterodimeric form. This chimaeric antibody has very high affinity and specificity, similar to that of the original mouse monoclonal antibody. PMID- 16253116 TI - Minimal features of paxillin that are required for the tyrosine phosphorylation of focal adhesion kinase. AB - Tyrosine phosphorylation of FAK (focal adhesion kinase) regulates signalling that results from the interaction of integrins with extracellular matrix and growth factor receptors. A critical step in this process is the phosphorylation of Tyr397 of FAK, which creates a binding site for Src family kinases, PI3K (phosphoinositide 3-kinase) and Shc (Src homology and collagen homology). An intact Tyr397 site is required for FAK-mediated regulation of cell migration, survival signals and full responsiveness to soluble growth factors. We showed previously that the adaptor protein paxillin is required for the overall tyrosine phosphorylation of FAK in embryonic stem cells [Wade, Bohl and Vande Pol (2002) Oncogene 21, 96-107]. In the present paper, we identify the minimal structural features of paxillin that are required to support overall FAK tyrosine phosphorylation and Tyr397 phosphorylation. Paxillin contains N-terminal leucine rich LD motifs that bind directly to FAK and four LIM (Lin-11, Isl-1 and Mec-3) domains in the C-terminus. We show that paxillin LIM domains 1, 2 and 3 are each required for FAK tyrosine phosphorylation, while LIM4 is dispensable. In addition to paxillin LIM domains 1, 2 and 3, a single LD motif on paxillin is required to support FAK tyrosine phosphorylation in embryonic stem cells. Both sequence and spatial requirements exist for LD motifs to support FAK tyrosine phosphorylation. Interestingly, synthetic LD motifs that fail to bind FAK in vitro are able to fully support FAK tyrosine phosphorylation, indicating that minimal interactions of LD motifs with FAK suffice. Our results demonstrate at least four distinct structural domains of paxillin support at least three distinct functions that are each required for FAK tyrosine phosphorylation. PMID- 16253118 TI - The differential role of Smad2 and Smad3 in the regulation of pro-fibrotic TGFbeta1 responses in human proximal-tubule epithelial cells. AB - In chronic renal diseases, progressive loss of renal function correlates with advancing tubulo-interstitial fibrosis. TGFbeta1-Smad (transforming growth factor beta1-Sma and Mad protein) signalling plays an important role in the development of renal tubulo-interstitial fibrosis. Secretion of CTGF (connective-tissue growth factor; CCN2) by PTECs (proximal-tubule epithelial cells) and EMT (epithelial-mesenchymal transdifferentiation) of PTECs to myofibroblasts in response to TGFbeta are critical Smad-dependent events in the development of tubulo-interstitial fibrosis. In the present study we have investigated the distinct contributions of Smad2 and Smad3 to expression of CTGF, E-cadherin, alpha-SMA (alpha-smooth-muscle actin) and MMP-2 (matrix-metalloproteinase-2) in response to TGFbeta1 treatment in an in vitro culture model of HKC-8 (transformed human PTECs). RNA interference was used to achieve selective and specific knockdown of Smad2 and Smad3. Cellular E-cadherin, alpha-SMA as well as secreted CTGF and MMP-2 were assessed by Western immunoblotting. TGFbeta1 treatment induced a fibrotic phenotype with increased expression of CTGF, MMP-2 and alpha SMA, and decreased expression of E-cadherin. TGFbeta1-induced increases in CTGF and decreases in E-cadherin expression were Smad3-dependent, whereas increases in MMP-2 expression were Smad2-dependent. Increases in alpha-SMA expression were dependent on both Smad2 and Smad3 and were abolished by combined knockdown of both Smad2 and Smad3. In conclusion, we have demonstrated distinct roles for Smad2 and Smad3 in TGFbeta1-induced CTGF expression and markers of EMT in human PTECs. This can be of therapeutic value in designing targeted anti-fibrotic therapies for tubulo-interstitial fibrosis. PMID- 16253119 TI - Adjuvant effect of synthetic oligodeoxyribonucleotides (CpG-ODN) from the Paracoccidioides brasiliensis gp43 gene on the Th2-Th1 immunomodulation of experimental paracoccidioidomycosis. AB - Paracoccidioidomycosis (PCM) is caused by the dimorphic fungus Paracoccidioides brasiliensis. Immunostimulatory effects of P. brasiliensis DNA and CpG oligodeoxyribonucleotides (CpG-ODN) have shown a Th2-Th1 immunomodulation of the isogenic murine model of susceptibility, which develops a progressive and disseminating disease. In this study, we investigated the optimum time interval and doses of CpG-ODN which are able to induce Th2-Th1 immunomodulation. The optimum concentrations for the induction of a decrease in antibody production were 0.5 and 1 microg. Mice immunized twice with CpG-ODN and gp43 (5 and 7 days before the challenge) showed a 60% higher chance of survival compared with the control group (nonimmunized), and an increase in Th1 isotype (IgG2a) was also observed. In vitro assays of naive and preimmunized mice showed discrete cellular proliferation when stimulated by suitable concentrations of CpG-ODN. Type 1 cytokines interleukin-12 (IL-12) and interferon-gamma were increased in cell culture supernatants, but no significant difference was found in Th2 IL-4 cytokines in stimulated or nonstimulated cell cultures. Concerning the Th2-Th1 kinetics in experimental PCM models by adjuvant effect of CpG-ODN, there are still many questions to be answered and clarified. However, the gathering of data obtained in this investigation has led us to suggest that the modulation of Th2 Th1 in experimental PCM depends on time and CpG-ODN concentration. PMID- 16253120 TI - Interleukin-2-activated natural killer cells may have a direct role in the control of Leishmania (Leishmania) amazonensis promastigote and macrophage infection. AB - To study the role of Natural Killer (NK) cells in Leishmania infection, peritoneal macrophages from BALB/c mice were infected with Leishmania (Leishmania) amazonensis promastigotes and incubated with interleukin-2 (IL-2) activated NK (A-NK) cells at different ratios of A-NK cells to infected macrophages (5:1, 1:1, 0.2:1). The A-NK cells were added either together with the parasites (0-h group) or 24 h later (24-h group). Morphological studies of the cultures revealed predominance of parasitic debris within macrophages that were in close contact with A-NK cells and the decrease in parasite recovery was directly proportional to the A-NK cell concentration used. Interferon-gamma (IFN gamma) and IL-12 were detected in the supernatant at levels proportional to the A NK cell concentration used. No significant difference was observed between the groups with respect to NO levels in the culture supernatant. When A-NK cells were added directly to the L. (L.) amazonensis promastigote cultures, the parasite recovery decreased proportional to the number of A-NK cells added. In vivo studies demonstrated smaller lesion sizes in animals inoculated with both parasites and A-NK cells compared with parasites alone. Histopathology of the skin lesions from animals receiving A-NK cells together with the parasites showed moderate parasitism and a nodular inflammatory infiltrate formed by mononuclear cells and a few vacuolized macrophages. In contrast, animals inoculated only with the parasites showed a highly parasitized dermis with infiltration of intensely vacuolized macrophages. These results demonstrate the role of A-NK cells in parasite lysis and in resistance of macrophages to L. (L.) amazonensis in the early phase of infection. PMID- 16253121 TI - Two doses of parenterally administered split influenza virus vaccine elicited high serum IgG concentrations which effectively limited viral shedding upon challenge in mice. AB - We have previously found that whole influenza virus vaccine induced a more rapid and stronger humoral response, particularly after the first dose of vaccine, than split virus vaccine in mice. In this study, we have evaluated the protective efficacy of whole and split influenza virus vaccines in mice using a nonlethal upper respiratory tract challenge model. We have also investigated the immunological correlates associated with no or very little viral shedding after viral challenge. Vaccination resulted in reduced viral shedding and shortened the duration of infection by at least 2 days. After one dose of vaccine, whole virus vaccine generally resulted in less viral shedding than split virus vaccine. In contrast, two doses of split virus vaccine, particularly the highest vaccine strengths of 15 and 30 microg HA, most effectively limited viral replication and these mice had high concentrations of prechallenge influenza-specific serum IgG. The vaccine formulation influenced the IgG2a/IgG1 ratio, and this IgG subclass profile was maintained upon challenge to some extent, although it did not influence the level of viral shedding. The concentration of postvaccination serum IgG showed an inverse relationship with the level of viral shedding after viral challenge. Therefore, serum IgG is an important factor in limiting viral replication in the upper respiratory tract upon challenge of an antigenically similar virus. PMID- 16253122 TI - Human IgG but not IgM antibodies can protect mice from the challenge with live O6 Escherichia coli. AB - We evaluated the ability of human anti-lipopolysaccharide (LPS) O6 immunoglobulin G (IgG) and IgM antibodies to protect mice challenged with Escherichia coli serotype O6:K2ac. Purified whole IgG, commercial gammaglobulin, whole IgM effluent, pool of normal human serum (NHS), agammaglobulinaemic serum (test groups) or phosphate-buffered saline (control group) was injected into adult male 18 h before a challenge with viable O6 E. coli. The mortality rate was assessed over a period of 72 h. To determine the opsonic and phagocytic activity of the antibody isotypes, we incubated peritoneal macrophages from the control and test groups collected at different times after challenge with the live bacteria with acridine orange for fluorescent analysis. Tumour necrosis factor (TNF)-alpha and interleukin (IL)-6 were quantified in serum of both the test and control groups. All mice that received commercial gammaglobulin or NHS survived. Purified whole IgG (containing 1.1 mg/l of anti-LPS O6 IgG antibodies) protected 87.5% of the animals tested in this experiment, while whole IgM-enriched effluent with 1.5 mg/l of anti-LPS O6 IgM antibodies protected only 12.5%. The agamma serum showed no protective capacity compared with PBS (serving as control). The minimal concentration of anti-LPS O6 IgG antibodies able to protect 50% of animals was 0.137 mg/l of purified whole IgG. Whole IgM-enriched effluent showed no protective capacity independently of the concentration tested (0.048-17.0 mg/l of anti-LPS O6 IgM antibodies). Fluorescent analysis of peritoneal macrophages from animals pretreated with purified whole IgG showed no bacteria at 8 h after the challenge. By contrast, whole IgM effluent showed an increasing number of live bacteria at the same time. Mice that had received whole IgM effluent (1.5 mg/l of anti-LPS O6 IgM antibodies) before the challenge with LPS O6 presented 20.5 microg/l of IL-6 and 1.5 microg/l of TNF-alpha. Serum from animals pretreated with purified IgG did not present any detectable pro-inflammatory cytokine. Our findings suggest that IgG but not IgM antibodies protect animals from a challenge with E. coli O6 serotype. PMID- 16253123 TI - Activation of natural killer-like YT-INDY cells by oligodeoxynucleotides and binding by homologous pattern recognition proteins. AB - The present study was designed to examine the binding and signalling effects of single base and CpG dinucleotide phosphodiester (Po) oligodeoxynucleotides (ODN) on the human natural killer (NK)-like cell line (YT-INDY). Single base Po ODN composed of 20-mers of guanosine (dG20), adenosine (dA20), cytosine (dC20) or thymidine (dT20) as well as 'conventional' Po CpG ODN were examined for their ability to bind and activate YT-INDY cells. Binding by dG20 and CpG ODN to YT INDY cells was saturable and specific. dG20 binding was competitively inhibited by homologous dG20 and heterologous CpG ODN but not by dC20 and dA20. Two different YT-INDY membrane proteins (18 and 29 kDa) were identified by ligand (Southwestern) blotting with biotinylated dG20 and CpG. The specificity of the ODN-binding protein(s) was further confirmed by ODN depletion experiments using a teleost recombinant protein orthologue [nonspecific cytotoxic cells (NCC) cationic antimicrobial protein-1 (ncamp-1)] known to bind CpG and dG20. Cell proliferation and activation studies showed that dG20 and CpG treatment of YT INDY cells induced cellular DNA synthesis (i.e. G1 to S-phase conversion). This signalling function was accompanied in dG20-treated cells by proliferation 10 h posttreatment. Both dG20 and CpG ODN binding induced a calcium flux in YT-INDY cells within seconds of treatment. These experiments demonstrated that Po single base dG20 and CpG ODN bind to a (potential) new class of cell-surface proteins that mediate the activation of YT-INDY cells. PMID- 16253124 TI - DNA vaccination using bacillus Calmette-Guerin-DNA as an adjuvant to enhance immune response to three kinds of swine diseases. AB - In order to enhance the immune efficacy of DNA vaccination, experiments were conducted to investigate the regulating effects of Bacillus Calmette-Guerin (BCG) DNA as an adjuvant on immune responses of mice against foot-and-mouth disease (FMD), Aujeszky's disease (AjD) and classical swine fever (CSF). BCG-DNA was purified from BCG by ion-exchange chromatography. Three DNA vaccines (pVSG, pVgD and pVE2) against the respective infection were constructed, and BCG-DNA was coimmunized to mice by muscle injection. The results showed that titres of specific immunoglobulin (Ig)G to the vaccines mounted remarkably in the sera of the adjuvant covaccinated mice (P < 0.01). Antibody isotype IgG2a and IgG1 also increased, respectively, in mice coimmunized with BCG-DNA compared with those of the control groups (P < 0.01). Cellular immune cytokine interferon-gamma and cytotoxic T lymphocytes were detected in coimmunized BCG-DNA groups (P < 0.05). Whereas interleukin-4, humoral immune cytokine, was not significant (P > 0.05). These results suggest that codelivery of BCG-DNA with DNA vaccines against FMD, AjD and CSF can enhance the induction of antigen-specific, especially, cell mediated immunity. PMID- 16253125 TI - Effect of rolipram, a phosphodiesterase IV inhibitor, on allergic footpad swelling using various adjuvants in mice. AB - We studied the effect of rolipram, a phosphodiesterase (PDE) IV inhibitor, on allergic footpad swelling in mice. For this study, varying adjuvants including complete Freund's adjuvant (CFA), incomplete Freund's adjuvant (IFA) and Imject Alum (Alum) were used because the extent of antigen-specifically induced T helper type 1 (Th1) and Th2 responses had been shown to depend on adjuvants used. To induce allergic footpad swelling, we immunized mice with ovalbumin (OVA) emulsified in either CFA or IFA, dissolved in Alum or in phosphate-buffered saline (PBS) as a control (day 0), followed by subcutaneous injection of the antigen into footpads on day 21. Rolipram was given orally to the animals daily from days 0-20. Results showed that treatment with rolipram was followed by an increase in early swelling at 0.5 h and a decrease in late swelling at 6 and 24 h in the CFA group. In the IFA group, rolipram significantly enhanced swelling at, but not after, 30 min. In the Alum and the PBS groups, the PDE inhibitor failed to affect the OVA-specific footpad reaction at all times examined. Treatment of the CFA and IFA groups with rolipram significantly inhibited the production of the Th1 antibody anti-OVA immunoglobulin G2a (IgG2a), and the drug enhanced Th2 cell-dependent anti-OVA IgE production. In both groups, rolipram also enhanced the secretion of Th2 cytokines including interleukin-4 (IL-4) and IL-10. These findings suggest that rolipram may facilitate early allergic footpad swelling mediated by Th2 immune responses, while the late phase of swelling associated with Th1 responses may be attenuated by the PDE IV inhibitor. PMID- 16253126 TI - The immunogenicity of a conformationally restricted peptide mimetic of meningococcal lipooligosaccharide. AB - Life-threatening meningitis and septicaemia caused by Neisseria meningitidis are a public health priority, and their prevention by vaccination is a major objective. Meningococcal capsular polysaccharide-based vaccines are effective against the major invasive serogroups, except for serogroup B, the capsule of which mimics human polysaccharides and is poorly immunogenic. An alternative vaccine candidate that has the potential to offer cross-protection against antigenically diverse meningococci is the lipooligosaccharide (LOS). The structurally constrained peptide mimetic, C22, of a bactericidal antibody epitope within LOS was previously shown to elicit cross-reactive antibodies to meningococcal LOS when complexed to NeutrAvidintrade mark as a carrier protein. The immunogenicity of this antigen in H-2(d) (BALB/c) and H-2(k) (C3H/HeN) haplotype mice was further investigated. Anti-LOS immunoglobulin G (IgG) antibody titres increased with the vaccine dose and correlated with the anti-C22 peptide antibody titres in both haplotypes. Antigen-stimulated Th1/Th2 cytokine secretion by splenocytes and antibody isotypes indicated a Th2-type immune response with IgG1 antibodies and a low titre of IgG2b. There was no serum bactericidal activity observed against the meningococcus. PMID- 16253127 TI - An extract of the mushroom Agaricus blazei Murill administered orally protects against systemic Streptococcus pneumoniae infection in mice. AB - The aim was to investigate the antibacterial effect of the biologically active and edible mushroom Agaricus blazei Murill (AbM). A water extract of AbM or PBS control was administered orally before or with challenge to NIH/OlaHsd mice, experimentally infected intraperitoneally with the moderately virulent Streptococcus pneumoniae serotype 6B. End points were bacteraemia and survival rate. The AbM extract, protected against systemic S. pneumoniae 6B infection in the mice. It was most effective when given 24 h before inoculation but did also have protective effects when given together with challenge compared with control. The lack of antibiotic effect on pneumococci in vitro and increased levels of cytokines MIP-2 and TNF-alpha in the serum of mice receiving AbM extract, indicated that the protective effect of AbM was due to the involvement of the native immune system. This is the first report of anti-infection effects of AbM in vivo. Our results suggest that AbM extract may be useful as additional prophylactic and possibly therapeutic treatment against bacterial and possibly other infections in humans. PMID- 16253128 TI - The impact of electrical charge on the viability and physiology of dendritic cells. AB - The use of electrical charge for electroporation or electrofusion is widely applied to customize dendritic cells (DC) and their immunological properties as anticancer vaccines. The aim of this study was to evaluate the influence of various electrical field strengths on the recovery, viability and physiology of DC. Immature DC were transferred into low-conductive medium and electrically charged within a range of 0-1500 V/cm. Viability was assessed by Trypan Blue dye exclusion or staining with impermeant nucleic acid stains and fluorescence activated cell sorter analysis. Additionally, apoptosis was determined by flow cytometry after staining with Annexin-V, endocytosis by uptake of fluorescein isothiocyanate-dextran and metabolic activity by a standardized fluorescent live/dead assay. There was a strong correlation between the electrical field strength and the viability and physiology of DC. Field strengths > or =1000 V/cm significantly impaired viability, metabolism and endocytotic activity. Dual fluorescence with 7-7-amino-actinomycin D and Annexin-V demonstrated that loss of viability was predominantly due to necrosis rather than apoptosis. Field strengths < or =500 V/cm allowed to maintain good cell viability and recovery of DC and did not cause alterations of metabolism and endocytosis. Therefore, the frequently used amplification of field strengths to improve the efficacy of electroporation and electrofusion requires critical re-evaluation. PMID- 16253129 TI - Tissue localization of Toll-like receptors in biopsy specimens of liver from children infected with hepatitis C virus. AB - Toll-like receptors (TLR) are important tools of innate immunity, localized mainly on cells of the immune system, but also have been shown on cells of other origin. In the current study, they have been searched in biopsy specimens of liver from children bearing chronic viral hepatitis of C type (HCV). TLR2, TLR3 and TLR4 were traced by means of polyclonal antibodies and avidin-biotin complex (ABC) immunohistochemistry. Besides, mRNA for TLR was looked for using specific primers and polymerase chain reaction. Several controls, including neutralization of primary antibody with respective blocking peptide, confirmed the specificity of the immunohistochemical reaction. All TLR tested could be visualized in a focal distribution in single hepatocytes and some cells of inflammatory infiltrates. There was no reaction whatsoever in liver samples not infected with hepatotropic virus. In molecular studies, mRNA for TLR2 and TLR4 was detected in both noninfected and hepatitis B virus-infected established cell lines of human hepatoma as well as in HCV(+) biopsy samples. These data indicate that TLR can be traced in liver cells, both at the protein and at the mRNA level. Their irregular and focal distribution in HCV(+), but not in HCV(-), liver suggests some role of TLR in the pathogenesis of chronic viral hepatitis, at least in children. PMID- 16253130 TI - Roles of group IIA phospholipase A2 and complement in killing of bacteria by acute phase serum. AB - The complement system is regarded as an important component of the innate defence system against invading bacteria. However, synergistic actions between the complement and the other components of innate immunity are incompletely known. Human group IIA phospholipase A(2) (hGIIA PLA(2)) is an effective antibacterial enzyme in serum of patients with severe bacterial infections. Our aim was to investigate the significance of complement and hGIIA PLA(2) in acute phase serum. Serum samples were collected from patients with acute bacterial infections and from healthy control subjects. We prepared hGIIA PLA(2)-depleted serum by immunoadsorption and inhibited the activity of complement by a specific inhibitor, compstatin. The bactericidal effects of treated and untreated serum were compared by incubating Staphylococcus aureus and Listeria monocytogenes in the presence of serum. Acute phase serum effectively killed S. aureus and L. monocytogenes, and depletion of hGIIA PLA(2) significantly reduced the antibacterial effect. Complement had a weak bactericidal effect against L. monocytogenes. We conclude that hGIIA PLA(2) is the major antibacterial factor in human acute phase serum against the gram-positive bacteria S. aureus and L. monocytogenes, exceeding complement in efficiency. PMID- 16253132 TI - "There's no place like home" a pilot study of perspectives of international health and social care professionals working in the UK. AB - BACKGROUND: Many countries are reporting health workforce shortages across a range of professions at a time of relatively high workforce mobility. Utilising the global market to supply shortage health skills is now a common recruitment strategy in many developed countries. At the same time a number of countries report a 'brain drain' resulting from professional people leaving home to work overseas. Many health and social care professionals make their way to the UK from other countries. This pilot study utilises a novel 'e-survey' approach to explore the motives, experiences and perspectives of non-UK health and social care professionals who were working or had worked in the UK. The study aims to understand the contributions of international health and social care workers to the UK and their 'home' countries. The purpose of the pilot study is also in part to test the appropriateness of this methodology for undertaking a wider study. RESULTS: A 24-item questionnaire with open-ended and multiple choice questions was circulated via email to 10 contacts who were from a country outside the UK, had trained outside the UK and had email access. These contacts were requested to forward the email to other contacts who met these criteria (and so on). The email was circulated over a one month pilot period to 34 contacts. Responses were from physiotherapists (n = 11), speech therapists (n = 4), social workers (n = 10), an occupational therapist (n = 1), podiatrists (n = 5), and others (n = 3). Participants were from Australia (n = 20), South Africa (n = 10), New Zealand (n = 3) and the Republic of Ireland (n = 1). Motives for relocating to the UK included travel, money and career opportunities. Participants identified a number of advantages and disadvantages of working in the UK compared to working in their home country health system. Respondents generally reported that by working in the UK, they had accumulated skills and knowledge that would allow them to contribute more to their profession and health system on their return home. CONCLUSION: This pilot study highlights a range of issues and future research questions for international learning and comparison for the health and social care professions as a result of international workforce mobility. The study also highlights the usefulness of an e-survey technique for capturing information from a geographically diverse and mobile group of professionals. PMID- 16253133 TI - A framework to evaluate research capacity building in health care. AB - BACKGROUND: Building research capacity in health services has been recognised internationally as important in order to produce a sound evidence base for decision-making in policy and practice. Activities to increase research capacity for, within, and by practice include initiatives to support individuals and teams, organisations and networks. Little has been discussed or concluded about how to measure the effectiveness of research capacity building (RCB) DISCUSSION: This article attempts to develop the debate on measuring RCB. It highlights that traditional outcomes of publications in peer reviewed journals and successful grant applications may be important outcomes to measure, but they may not address all the relevant issues to highlight progress, especially amongst novice researchers. They do not capture factors that contribute to developing an environment to support capacity development, or on measuring the usefulness or the 'social impact' of research, or on professional outcomes. The paper suggests a framework for planning change and measuring progress, based on six principles of RCB, which have been generated through the analysis of the literature, policy documents, empirical studies, and the experience of one Research and Development Support Unit in the UK. These principles are that RCB should: develop skills and confidence, support linkages and partnerships, ensure the research is 'close to practice', develop appropriate dissemination, invest in infrastructure, and build elements of sustainability and continuity. It is suggested that each principle operates at individual, team, organisation and supra-organisational levels. Some criteria for measuring progress are also given. SUMMARY: This paper highlights the need to identify ways of measuring RCB. It points out the limitations of current measurements that exist in the literature, and proposes a framework for measuring progress, which may form the basis of comparison of RCB activities. In this way it could contribute to establishing the effectiveness of these interventions, and establishing a knowledge base to inform the science of RCB. PMID- 16253134 TI - Reliability of old and new ventricular fibrillation detection algorithms for automated external defibrillators. AB - BACKGROUND: A pivotal component in automated external defibrillators (AEDs) is the detection of ventricular fibrillation by means of appropriate detection algorithms. In scientific literature there exists a wide variety of methods and ideas for handling this task. These algorithms should have a high detection quality, be easily implementable, and work in real time in an AED. Testing of these algorithms should be done by using a large amount of annotated data under equal conditions. METHODS: For our investigation we simulated a continuous analysis by selecting the data in steps of one second without any preselection. We used the complete BIH-MIT arrhythmia database, the CU database, and the files 7001-8210 of the AHA database. All algorithms were tested under equal conditions. RESULTS: For 5 well-known standard and 5 new ventricular fibrillation detection algorithms we calculated the sensitivity, specificity, and the area under their receiver operating characteristic. In addition, two QRS detection algorithms were included. These results are based on approximately 330,000 decisions (per algorithm). CONCLUSION: Our values for sensitivity and specificity differ from earlier investigations since we used no preselection. The best algorithm is a new one, presented here for the first time. PMID- 16253135 TI - Age as a determinant of nutritional status: a cross sectional study. AB - BACKGROUND: Undenutrition is known to be prevalent and largely unrecognised in older patients; however, aberrations in indicators of nutritional status may simply reflect effects of age and/or functional disability. OBJECTIVE: The aim of this study was to measure the effect, if any of age on nutritional status in older patients. DESIGN: 445 randomly selected hospitalised patients consented to nutritional status assessment derived from anthropometric, haematological, and biochemical data within 72 hours of admission. Nutritional status was compared between those age < 75 years and those aged 75 years or more. Using multiple regression models, we measured the association between age and nutritional assessment variables after adjusting for disability, chronic illness, medications, smoking and tissue inflammation. RESULTS: Body weight, body mass index, mid-upper arm circumference, haemoglobin, serum albumin and plasma ascorbic acid were all significantly lower in people aged > or = 75 years compared with those < 75 years of age. Although riboflavin (vitamin B2), 25OH VitD3, red-cell folate and vitamin B12 concentrations were lower in those aged > or = 75 years, differences were not statistically significant. After adjusting for disability and co-morbidity in a multivariate analysis, age alone had a significant and independent effect on important anthropometric and biochemical nutritional assessment variables. CONCLUSION: Increasing age is independently associated with poor nutritional status. This may partly explain the poor clinical outcome in older patients. PMID- 16253136 TI - Influence of hypoxia on the domiciliation of mesenchymal stem cells after infusion into rats: possibilities of targeting pulmonary artery remodeling via cells therapies? AB - BACKGROUND: Bone marrow (BM) cells are promising tools for vascular therapies. Here, we focused on the possibility of targeting the hypoxia-induced pulmonary artery hypertension remodeling with systemic delivery of BM-derived mesenchymal stem cells (MSCs) into non-irradiated rats. METHODS: Six-week-old Wistar rats were exposed to 3-week chronic hypoxia leading to pulmonary artery wall remodeling. Domiciliation of adhesive BM-derived CD45- CD73+ CD90+ MSCs was first studied after a single intravenous infusion of Indium-111-labeled MSCs followed by whole body scintigraphies and autoradiographies of different harvested organs. In a second set of experiments, enhanced-GFP labeling allowed to observe distribution at later times using sequential infusions during the 3-week hypoxia exposure. RESULTS: A 30% pulmonary retention was observed by scintigraphies and no differences were observed in the global repartition between hypoxic and control groups. Intrapulmonary radioactivity repartition was homogenous in both groups, as shown by autoradiographies. BM-derived GFP-labeled MSCs were observed with a global repartition in liver, in spleen, in lung parenchyma and rarely in the adventitial layer of remodeled vessels. Furthermore this global repartition was not modified by hypoxia. Interestingly, these cells displayed in vivo bone marrow homing, proving a preservation of their viability and function. Bone marrow homing of GFP-labeled MSCs was increased in the hypoxic group. CONCLUSION: Adhesive BM-derived CD45- CD73+ CD90+ MSCs are not integrated in the pulmonary arteries remodeled media after repeated intravenous infusions in contrast to previously described in systemic vascular remodeling or with endothelial progenitor cells infusions. PMID- 16253137 TI - The effect of performance-related pay of hospital doctors on hospital behaviour: a case study from Shandong, China. AB - BACKGROUND: With the recognition that public hospitals are often productively inefficient, reforms have taken place worldwide to increase their administrative autonomy and financial responsibility. Reforms in China have been some of the most radical: the government budget for public hospitals was fixed, and hospitals had to rely on charges to fill their financing gap. Accompanying these changes was the widespread introduction of performance-related pay for hospital doctors- termed the "bonus" system. While the policy objective was to improve productivity and cost recovery, it is likely that the incentive to increase the quantity of care provided would operate regardless of whether the care was medically necessary. METHODS: The primary concerns of this study were to assess the effects of the bonus system on hospital revenue, cost recovery and productivity, and to explore whether various forms of bonus pay were associated with the provision of unnecessary care. The study drew on longitudinal data on revenue and productivity from six panel hospitals, and a detailed record review of 2303 tracer disease patients (1161 appendicitis patients and 1142 pneumonia patients) was used to identify unnecessary care. RESULTS: The study found that bonus system change over time contributed significantly to the increase in hospital service revenue and hospital cost recovery. There was an increase in unnecessary care and in the probability of admission when the bonus system switched from one with a weaker incentive to increase services to one with a stronger incentive, suggesting that improvement in the financial health of public hospitals was achieved at least in part through the provision of more unnecessary care and drugs and through admitting more patients. CONCLUSION: There was little evidence that the performance-related pay system as designed by the sample of Chinese public hospitals was socially desirable. Hospitals should be monitored more closely by the government, and regulations applied to limit opportunistic behaviour. Otherwise, the containment of government financing for public facilities may result in an increase in the provision of unnecessary care, an increase in health costs to society, and a waste in social resources. PMID- 16253138 TI - Dietary inulin intake and age can significantly affect intestinal absorption of calcium and magnesium in rats: a stable isotope approach. AB - BACKGROUND: previous studies have shown that non-digestible inulin-type fructan intake can increase intestinal mineral absorption in both humans and animals. However, this stimulatory effect on intestinal absorption may depend on experimental conditions such as duration of fermentable fiber intake, mineral diet levels and animals' physiological status, in particular their age. OBJECTIVES: the aim of this study was to determine the effect of inulin intake on Ca and Mg absorption in rats at different age stages. METHODS: eighty male Wistar rats of four different ages (2, 5, 10 and 20 months) were randomized into either a control group or a group receiving 3.75% inulin in their diet for 4 days and then 7.5% inulin for three weeks. The animals were fed fresh food and water ad libitum for the duration of the experiment. Intestinal absorption of Ca and Mg was determined by fecal monitoring using stable isotopic tracers. Ca and Mg status was also assessed. RESULTS: absorption of Ca and Mg was significantly lower in the aged rats (10 and 20 mo) than in the young and adult rat groups. As expected, inulin intake increased Ca and Mg absorption in all four rat groups. However, inulin had a numerically greater effect on Ca absorption in aged rats than in younger rats whereas its effect on Mg absorption remained similar across all four rat age groups. CONCLUSION: the extent of the stimulatory effect of inulin on absorption of Ca may differ according to animal ages. Further studies are required to explore this effect over longer inulin intake periods, and to confirm these results in humans. PMID- 16253139 TI - Dual role of TRBP in HIV replication and RNA interference: viral diversion of a cellular pathway or evasion from antiviral immunity? AB - Increasing evidence indicates that RNA interference (RNAi) may be used to provide antiviral immunity in mammalian cells. Human micro (mi)RNAs can inhibit the replication of a primate virus, whereas a virally-encoded miRNA from HIV inhibits its own replication. Indirect proof comes from RNAi suppressors encoded by mammalian viruses. Influenza NS1 and Vaccinia E3L proteins can inhibit RNAi in plants, insects and worms. HIV-1 Tat protein and Adenovirus VA RNAs act as RNAi suppressors in mammalian cells. Surprisingly, many RNAi suppressors are also inhibitors of the interferon (IFN)-induced protein kinase R (PKR) but the potential overlap between the RNAi and the IFN pathways remains to be determined. The link between RNAi as an immune response and the IFN pathway may be formed by a cellular protein, TRBP, which has a dual role in HIV replication and RNAi. TRBP has been isolated as an HIV-1 TAR RNA binding protein that increases HIV expression and replication by inhibiting PKR and by increasing translation of structured RNAs. A recent report published in the Journal of Virology shows that the poor replication of HIV in astrocytes is mainly due to a heightened PKR response that can be overcome by supplying TRBP exogenously. In two recent papers published in Nature and EMBO Reports, TRBP is now shown to interact with Dicer and to be required for RNAi mediated by small interfering (si) and micro (mi)RNAs. The apparent discrepancy between TRBP requirement in RNAi and in HIV replication opens the hypotheses that RNAi may be beneficial for HIV-1 replication or that HIV-1 may evade the RNAi restriction by diverting TRBP from Dicer and use it for its own benefit. PMID- 16253140 TI - Severe allergic reactions to guinea pig. AB - BACKGROUND: Allergic sensitization and reactions to guinea pig (Cavia porcellus) have been well documented in laboratory animal handlers, primarily manifesting as rhinitis, conjunctivitis, and asthma. Severe allergic reactions, however, are rare. METHODS: We report two patients with severe allergic reactions following non-occupational exposure to guinea pigs. The first patient, an 11-year-old female, developed ocular, nasal, skin and laryngeal edema symptoms immediately after handling a guinea pig. The second patient, a 24-year-old female, developed symptoms of isolated laryngeal edema after cleaning a guinea pig cage. Percutaneous skin testing, RAST, ELISA and ELISA inhibition testing with guinea pig extract were performed. RESULTS: Both patients had IgE-mediated allergy to guinea pig confirmed by ELISA and either RAST or skin testing. ELISA inhibition studies confirmed the specificity of the IgE reactivity to guinea pig. CONCLUSION: Severe IgE-mediated reactions can occur following non-occupational guinea pig exposure. Physicians should be aware of this possibility. PMID- 16253141 TI - Distribution of cytochrome P450 2C, 2E1, 3A4, and 3A5 in human colon mucosa. AB - BACKGROUND: Despite the fact that the alimentary tract is part of the body's first line of defense against orally ingested xenobiotica, little is known about the distribution and expression of cytochrome P450 (CYP) enzymes in human colon. Therefore, expression and protein levels of four representative CYPs (CYP2C(8), CYP2E1, CYP3A4, and CYP3A5) were determined in human colon mucosa biopsies obtained from ascending, descending and sigmoid colon. METHODS: Expression of CYP2C, CYP2E1, CYP3A4, and CYP3A5 mRNA in colon mucosa was determined by RT-PCR. Protein concentration of CYPs was determined using Western blot methods. RESULTS: Extensive interindividual variability was found for the expression of most of the genes. However, expression of CYP2C mRNA levels were significantly higher in the ascending colon than in the sigmoid colon. In contrast, mRNA levels of CYP2E1 and CYP3A5 were significantly lower in the ascending colon in comparison to the descending and sigmoid colon. In sigmoid colon protein levels of CYP2C8 were significantly higher by ~73% than in the descending colon. In contrast, protein concentration of CYP2E1 was significantly lower by ~81% in the sigmoid colon in comparison to the descending colon. CONCLUSION: The current data suggest that the expression of CYP2C, CYP2E1, and CYP3A5 varies in different parts of the colon. PMID- 16253142 TI - Using hexamers to predict cis-regulatory motifs in Drosophila. AB - BACKGROUND: Cis-regulatory modules (CRMs) are short stretches of DNA that help regulate gene expression in higher eukaryotes. They have been found up to 1 megabase away from the genes they regulate and can be located upstream, downstream, and even within their target genes. Due to the difficulty of finding CRMs using biological and computational techniques, even well-studied regulatory systems may contain CRMs that have not yet been discovered. RESULTS: We present a simple, efficient method (HexDiff) based only on hexamer frequencies of known CRMs and non-CRM sequence to predict novel CRMs in regulatory systems. On a data set of 16 gap and pair-rule genes containing 52 known CRMs, predictions made by HexDiff had a higher correlation with the known CRMs than several existing CRM prediction algorithms: Ahab, Cluster Buster, MSCAN, MCAST, and LWF. After combining the results of the different algorithms, 10 putative CRMs were identified and are strong candidates for future study. The hexamers used by HexDiff to distinguish between CRMs and non-CRM sequence were also analyzed and were shown to be enriched in regulatory elements. CONCLUSION: HexDiff provides an efficient and effective means for finding new CRMs based on known CRMs, rather than known binding sites. PMID- 16253143 TI - Clinical presentation and prognostic factors of Streptococcus pneumoniae meningitis according to the focus of infection. AB - BACKGROUND: We conducted a nationwide study in Denmark to identify clinical features and prognostic factors in patients with Streptococcus pneumoniae according to the focus of infection. METHODS: Based on a nationwide registration, clinical information's was prospectively collected from all reported cases of pneumococcal meningitis during a 2-year period (1999-2000). Clinical and laboratory findings at admission, clinical course and outcome of the disease including follow-up audiological examinations were collected retrospectively. The focus of infection was determined according to the clinical diagnosis made by the physicians and after review of the medical records. RESULTS: 187 consecutive cases with S. pneumoniae meningitis were included in the study. The most common focus was ear (30%), followed by lung (18%), sinus (8%), and other (2%). In 42% of cases a primary infection focus could not be determined. On admission, fever and an altered mental status were the most frequent findings (in 93% and 94% of cases, respectively), whereas back rigidity, headache and convulsion were found in 57%, 41% and 11% of cases, respectively. 21% of patients died during hospitalisation (adults: 27% vs. children: 2%, Fisher Exact Test, P < 0.001), and the causes of death were due to neurological- and systemic complications or the combination of both in 8%, 5% and 6% of cases, respectively. Other causes (e.g. gastrointestinal bleeding, incurable cancer) accounted for 2% of cases. 41% of survivors had neurological sequelae (hearing loss: 24%, focal neurological deficits: 16%, and the combination of both: 1%). The mortality varied with the focus of the infection (otogenic: 7%, sinusitic: 33%, pneumonic: 26%, other kind of focus: 50%, no primary infection focus: 21%, Log rank test: P = 0.0005). Prognostic factors associated with fatal outcome in univariate logistic regression analysis were advanced age, presence of an underlying disease, history of headache, presence of a lung focus, absence of an otogenic focus, having a CT scan prior to lumbar puncture, convulsions, requirement of assisted ventilation, and alterations in various CSF parameters (WBC < 500 cells/microL, high protein levels, glucose levels < 1 mmol/L, low CSF/blood glucose levels), P < 0.05. Independent prognostic factor associated with fatal outcome in multivariate logistic regression analysis was convulsions (OR: 4.53, 95%CI: (1.74-11.8), p = 0.002), whereas presence of an otogenic focus was independently associated with a better survival (OR: 6.09, 95%CI: (1.75-21.2), P = 0.005). CONCLUSION: These results emphasize the prognostic importance of an early recognition of a predisposing focus to pneumococcal meningitis. PMID- 16253145 TI - Cultural approach to HIV/AIDS harm reduction in Muslim countries. AB - Muslim countries, previously considered protected from HIV/AIDS due to religious and cultural norms, are facing a rapidly rising threat. Despite the evidence of an advancing epidemic, the usual response from the policy makers in Muslim countries, for protection against HIV infection, is a major focus on propagating abstention from illicit drug and sexual practices. Sexuality, considered a private matter, is a taboo topic for discussion. Harm reduction, a pragmatic approach for HIV prevention, is underutilized. The social stigma attached to HIV/AIDS, that exists in all societies is much more pronounced in Muslim cultures. This stigma prevents those at risk from coming forward for appropriate counseling, testing, and treatment, as it involves disclosure of risky practices. The purpose of this paper is to define the extent of the HIV/AIDS problem in Muslim countries, outline the major challenges to HIV/AIDS prevention and treatment, and discuss the concept of harm reduction, with a cultural approach, as a strategy to prevent further spread of the disease. Recommendations include integrating HIV prevention and treatment strategies within existing social, cultural and religious frameworks, working with religious leaders as key collaborators, and provision of appropriate healthcare resources and infrastructure for successful HIV prevention and treatment programs in Muslim countries. PMID- 16253144 TI - Limitations of mRNA amplification from small-size cell samples. AB - BACKGROUND: Global mRNA amplification has become a widely used approach to obtain gene expression profiles from limited material. An important concern is the reliable reflection of the starting material in the results obtained. This is especially important with extremely low quantities of input RNA where stochastic effects due to template dilution may be present. This aspect remains under documented in the literature, as quantitative measures of data reliability are most often lacking. To address this issue, we examined the sensitivity levels of each transcript in 3 different cell sample sizes. ANOVA analysis was used to estimate the overall effects of reduced input RNA in our experimental design. In order to estimate the validity of decreasing sample sizes, we examined the sensitivity levels of each transcript by applying a novel model-based method, TransCount. RESULTS: From expression data, TransCount provided estimates of absolute transcript concentrations in each examined sample. The results from TransCount were used to calculate the Pearson correlation coefficient between transcript concentrations for different sample sizes. The correlations were clearly transcript copy number dependent. A critical level was observed where stochastic fluctuations became significant. The analysis allowed us to pinpoint the gene specific number of transcript templates that defined the limit of reliability with respect to number of cells from that particular source. In the sample amplifying from 1000 cells, transcripts expressed with at least 121 transcripts/cell were statistically reliable and for 250 cells, the limit was 1806 transcripts/cell. Above these thresholds, correlation between our data sets was at acceptable values for reliable interpretation. CONCLUSION: These results imply that the reliability of any amplification experiment must be validated empirically to justify that any gene exists in sufficient quantity in the input material. This finding has important implications for any experiment where only extremely small samples such as single cell analyses or laser captured microdissected cells are available. PMID- 16253147 TI - Ventilation during cardiopulmonary resuscitation: the right tool for the job. PMID- 16253148 TI - Nitric oxide as a bactericidal agent: is the cure worse than the disease? PMID- 16253146 TI - The expression of HSP60 and HSP10 in large bowel carcinomas with lymph node metastase. AB - BACKGROUND: The involvement of Heat Shock Proteins (HSP) in cancer development and progression is a widely debated topic. The objective of the present study was to evaluate the presence and expression of HSP60 and HSP10 in a series of large bowel carcinomas and locoregional lymph nodes with and without metastases. METHODS: 82 Astler and Coller's stage C2 colorectal cancers, of which 48 well differentiated and 34 poorly-differentiated, were selected along with 661 lymph nodes, including 372 with metastases and 289 with reactive hyperplasia only, from the same tumours. Primitive tumours and both metastatic and reactive lymph nodes were studied; specifically, three different compartments of the lymph nodes, secondary follicle, paracortex and medullary sinus, were also analysed. An immunohistochemical research for HSP60 and HSP10 was performed and the semiquantitative results were analysed by statistical analysis to determine the correlation between HSPs expression and 1) tumour grading; 2) degree of inflammation; 3) number of lymph nodes involved; 4) lymph node compartment hyperplasia. Moreover, western blotting was performed on a smaller group of samples to confirm the immunohistochemical results. RESULTS: Our data show that the expression of HSP60, in both primary tumour and lymph node metastasis, is correlated with the tumoral grade, while the HSP10 expression is not. Nevertheless, the levels of HSP10 are commonly higher than the levels of HSP60. In addition, statistical analyses do not show any correlation between the degree of inflammation and the immunopositivity for both HSP60 and HSP10. Moreover, we find a significant correlation between the presence of lymph node metastases and the positivity for both HSP60 and HSP10. In particular, metastatic lymph nodes show a higher percentage of cells positive for both HSP60 and HSP10 in the secondary follicles, and for HSP10 in the medullary sinuses, when compared with hyperplastic lymph nodes. CONCLUSION: HSP60 and HSP10 may have diagnostic and prognostic significance in the management of this tumour and their overexpression in tumoral cells may be functionally related to tumoral progression. We hypothesise that their expression in follicular and medullary cells of lymph nodes may be induced by formation of metastases. Further studies based on these observations could lead to a better understanding of the HSPs involvement in colorectal cancer progression, as well as other neoplasms. PMID- 16253149 TI - Effects of expiratory rib-cage compression on oxygenation, ventilation, and airway-secretion removal in patients receiving mechanical ventilation. AB - BACKGROUND: Expiratory rib-cage compression, a chest physiotherapy technique, is well known as the "squeezing" technique in Japan. OBJECTIVE: To determine the effects of rib-cage compression on airway-secretion removal, oxygenation, and ventilation in patients receiving mechanical ventilation. SETTING: An intensive care unit of an emergency and critical care center at a tertiary-care teaching hospital in Tokyo, Japan. METHODS: Thirty-one intubated, mechanically ventilated patients in an intensive care unit were studied in a randomized, crossover trial. The patients received endotracheal suctioning with or without rib-cage compression, with a minimum 3-hour interval between the 2 interventions. Rib-cage compression was performed for 5 min before endotracheal suctioning. Arterial blood gas and respiratory mechanics were measured 5 min before endotracheal suctioning (baseline) and 25 min after suctioning. The 2 measurement periods were carried out on the same day. RESULTS: There were no significant differences in the ratio of arterial partial pressure of oxygen to fraction of inspired oxygen, P(aCO2), or dynamic compliance of the respiratory system between the 2 periods (before and after endotracheal suctioning). Moreover, there were no significant differences in airway-secretion removal between the 2 periods. CONCLUSIONS: This study suggests that rib-cage compression prior to endotracheal suctioning does not improve airway-secretion removal, oxygenation, or ventilation after endotracheal suctioning in this unselected population of mechanically ventilated patients. PMID- 16253150 TI - Positive expiratory pressure changes aerosol distribution in patients with cystic fibrosis. AB - HYPOTHESIS: We hypothesized that aerosol distribution in the lungs of patients with cystic fibrosis changes with positive expiratory pressure (PEP). METHODS: Eight patients were randomized to one of 2 conditions. On one study day, patients inhaled saline aerosol containing 99mtechnetium generated by a Pari LC Plus nebulizer and exhaled through a Pari PEP device. On another day, the same patients exhaled through a low-resistance Pari filter (no PEP). Afterwards, they underwent gamma-camera lung imaging. Images were analyzed for lung deposition fraction, expressed as a percent of the initial nebulizer activity, and deposition pattern, expressed in terms of inner-outer and apical-basal ratios. RESULTS: Lung deposition fraction was significantly lower with the Pari PEP device; the mean + SD deposition fraction was 6.10 + 3.05% (median 6.20%) with PEP, compared to 10.76 + 4.52% (median 10.32%) (p = 0.0078) without PEP. The inner-outer ratio was 2.01 + 0.69 (median 2.23) with PEP, which was significantly lower than without PEP (2.76 + 1.33, median 2.55) (p = 0.004). The apical-basal ratio was 0.82 + 0.31 (median 0.80) with PEP, which was not significantly different from no PEP (1.00 + 0.49, median 0.90). CONCLUSION: These results indicate that less aerosol is deposited in the lungs of patients with cystic fibrosis when the Pari LC Plus nebulizer is used with the Pari PEP device, as described in these experiments. Nevertheless, aerosol administration with this nebulizer and PEP device also results in a proportional redistribution of aerosol to the peripheral airways, compared to nebulization without the PEP device. The clinical relevance of this subtle redistribution of aerosol in cystic fibrosis patients will probably depend on the drug administered and disease severity. PMID- 16253151 TI - Comparison of an oxygen-powered flow-limited resuscitator to manual ventilation with an adult 1,000-mL self-inflating bag. AB - BACKGROUND: Positive-pressure ventilation of patients with unprotected airways during cardiopulmonary resuscitation can cause gastric dilation. OBJECTIVE: Determine if there is a significant difference in volume delivered to lungs and stomach while using an adult 1,000-mL disposable bag-valve-mask (BVM) device and the oxygen-powered, flow-limited Oxylator EMX resuscitator. METHODS: We used a bench model to simulate a patient with an unprotected airway, consisting of an intubation manikin, lung analog, and simulated lower esophageal sphincter set at an opening pressure of 20 cm H2O. The BVM and the Oxylator were used to provide mask ventilation at a verbally prompted rate of 12 breaths/min. RESULTS: The volumes delivered with the BVM and the Oxylator to the lungs and stomach were not significantly different: 262 + 112 mL versus 297 + 99 mL and 227 + 199 mL versus 159 + 73 mL, respectively. CONCLUSION: Our study found no significant difference between the Oxylator and BVM when comparing tidal volume delivered to lungs and stomach during ventilation of a simulated unconscious nonintubated patient. More research on BVM use and the Oxylator should be done to validate the American Heart Association's guideline recommendations for ventilating unconscious patients with unprotected airways. Research on gastric dilation during cardiopulmonary resuscitation needs to be done with bench models using manikins that simulate chest excursion, bidirectional airway flow, lung impedance, and gastric compliance. PMID- 16253153 TI - An evaluation of home volume ventilators that support open-circuit mouthpiece ventilation. AB - BACKGROUND: Open-circuit mouthpiece ventilation (MPV) is a form of noninvasive ventilation that can be used to provide portable daytime ventilatory support for neuromuscular patients with chronic respiratory failure. MPV has been reported to reduce the risk of respiratory infection due to tracheostomy, and to improve cough and voice function and patient quality of life. Despite these potential benefits, mouthpiece ventilation is not widely used. This may be due in part to the fact that little information is available as to which ventilators can support this application. OBJECTIVE: To determine which volume-cycled portable home ventilators currently available in the United States will support MPV, and what peak inspiratory flow rates create adequate circuit pressure to prevent low pressure alarming. METHODS: We used a commercially available MPV breathing circuit with a set tidal volume range of 500?N1,000 mL with each of 8 ventilators currently available in the United States. RESULTS: Six of the 8 ventilators supported MPV: Respironics Lifecare PLV-100 and PLV Continuum, Mallinckrodt Achieva PSO2, Pulmonetics LTV800, Newport HT50, and Uni-Vent Eagle 754. PMID- 16253152 TI - The antimicrobial effect of nitric oxide on the bacteria that cause nosocomial pneumonia in mechanically ventilated patients in the intensive care unit. AB - BACKGROUND: Nosocomial pneumonia is the second most frequent nosocomial infection and the leading cause of death from hospital-acquired infection. Endogenously produced nitric oxide is an important component of the body's natural defense mechanism. Recent studies have demonstrated that exogenous gaseous nitric oxide (gNO) is bactericidal and that inhaled gNO is beneficial to bacterial clearance. OBJECTIVE: Determine the antimicrobial effect of exogenous gNO in vitro against organisms from culture collections and pathogens derived from tracheal aspirates of mechanically ventilated patients with pneumonia in an intensive care unit. METHODS: Using bacterial isolates in pure culture, a 0.5 McFarland standard (10(8) colony-forming-units [cfu] per mL) was prepared and further diluted to 1:1,000 with saline, to 10(5) cfu/mL. For each isolate tested, 3 mL was pipetted into each well of a 6-well plate, and placed in a specially designed incubator with compartments for both a treatment arm and a control arm. Both chambers received a continuous flow of heated, humidified gas. The treatment chamber had 200 ppm of gNO in the gas flow, which is higher than the clinically accepted concentration for gNO. Samples were drawn off at time intervals, plated onto Columbia agar base with 5% sheep blood, and placed in a traditional incubator at 35 degrees C for a minimum of 24 h. All tests were performed in duplicate. The colony-forming units were visually counted to determine percentage kill. RESULTS: There was total kill (100% of all colony-forming units) of each bacterial strain subjected to the test conditions at between 2 and 6 h of exposure to 200 ppm gNO. CONCLUSION: gNO is bactericidal against various strains of bacteria suspended in saline, including both Gram-positive and Gram-negative organisms, and those that commonly cause nosocomial pneumonia in mechanically ventilated patients. Future work should focus on developing strategies that maximize the antimicrobial effect while minimizing the effect of these same interventions on host cells. PMID- 16253154 TI - [The renin-angiotensin system in cardiovascular pathogenesis caused renal diseases: a new recognition]. PMID- 16253156 TI - [Antagonists of angiotensin receptors in hypertension therapy: function and status]. PMID- 16253158 TI - [Effects of angiotensin converting enzyme inhibitor with different doses on plasma brain natriuretic peptide and norepinephrine in patients with chronic heart failure]. AB - OBJECTIVE: To investigate the effects of angiotensin converting enzyme inhibitor (ACEI) with different doses on the plasma brain natriuretic peptide and norepinephrine (NE) of patients with chronic heart failure and the feasibility and safety of high dose ACEI treatment. METHODS: Sixty-six patients with chronic heart failure were randomly divided into 2 groups: low-dose group, treated by perindopril with the dosage of 2 mg/d initially and gradually increased up to 2-4 mg/d, and high-dose group, treated by perindopril with the dosage of 2 mg/d initially and gradually increased up to 8 approximately 10 mg/d. Treatment lasted for 12 weeks. Before and after the treatment peripheral blood was collected to test the plasma NE and brain natriuretic peptide levels, and the left ventricular ejection fraction (LVEF), left ventricular end diameter (LVED), blood pressure, and heart rate were examined. The differences of these indexes between the two groups and within the same group before and after the 12-week treatment were analyzed. Thirty healthy subjects were used as control group. RESULTS: The levels of plasma brain natriuretic peptide and NE of the chronic heart failure patients were significantly higher than those of the normal controls, and the higher the NYHA class of heart failure the higher the plasma levels of NE and brain natriuretic peptide. The plasma brain natriuretic peptide level was negatively correlated with LVEF (r = -0.327, P = 0.012) and positively correlated with plasma NE level (r = 0.402, P = 0.002) and LVED (r = 0.42, P = 0.015). The plasma brain natriuretic peptide and NE levels in the high-dose group were 6 microg/L +/ 4 microg/L and 250 ng/L +/- 63 ng/L respectively, both significantly lower than those of the low-dose group (8 microg/L +/- 4 microg/L and 387 ng/L +/- 211 ng/L respectively, both P < 0.05). Most of the patients in these 2 groups tolerated well. CONCLUSION: Plasma brain natriuretic peptide level is negatively correlated with the severity of heart failure and positively correlated with plasma NE. Treatment of CHF by perindopril with gradually increasing dose obviously decreases the plasma brain natriuretic peptide and NE levels and is tolerable for the patients. PMID- 16253159 TI - [Standard treatment of liver malignancies with radiofrequency ablation]. AB - OBJECTIVE: To investigate the role of standard treatment with ultrasound-guided radiofrequency ablation (RFA) in improving the treatment level of liver malignancies. METHODS: 302 patients with 476 liver malignancies were treated with established protocol and adjuvant measures and subjected to efficiency analysis. In the 302 patients, 181 had 282 hepatocellular carcinomas (HCC) with a mean diameter of 4.2 cm, and 121 had 194 metastatic liver carcinomas (MLC) with a mean diameter of 3.9 cm. According to UICC-TNM system 50 patients (27.6%) were in stage I/II and 131 (72.4%) in stage III/IV (including 39 patients with recurrent HCC after surgical resection). A standard protocol and an individualized protocol were used to treat the tumors based on their size, shape and special location such as the distance from diaphragm, gallbladder and gastrointestinal tract. Needle placement method and operation skill for the tumor region adjacent to important structures were described. Some adjuvant measures such as supplementary fine needle localization, local saline injection and feeding vessel ablation were used to improve RFA efficacy in tumors with different features. Local ablation of bleeding site and haemostatic administration systemically were adopted to deal with bleeding. For the patients with tumor adjacent to gastrointestinal tract, prolonged fasting after the RFA procedure was required. the patients were followed up regularly to assess the treatment efficiency, and the tumor was considered complete necrosed if no viability was found on enhanced CT or enhanced US one month after RFA. RESULTS: The tumor necrosis rate was 95.7% (270/282 tumors) for HCC, 94.8% (184/194 tumors) for MLC, 91.1% (51/56 tumors) for tumor near gastrointestinal tract, 88.5% (69/78 tumors) for tumors near diaphragm, and 94.3% (49/52 tumors) for tumor near gallbladder. The local recurrence rate was 10.3% (29/282 tumors) for HCC and 14.4% (28/194 tumors) for MLC. The 1, 2 and 3 year overall survival rates were 87.6%, 67.4% and 58.6% in the HCC patients, and 87.4%, 48.2%, 25.3% in the MLC patients respectively. The 1, 2 and 3 year survival rates of 50 HCC patients in early (I-II) stages were 90.7%, 85.9% and 73.7%, respectively. The incidence of complications was 2.2% (13/583 sessions), including 5 cases of hemorrhage, 1 case colon perforation, 8 cases of injury of adjacent structures. CONCLUSION: Application of proper protocol and adjuvant measures plays an important role in improving tumor ablation rate. Knowledge about possible complications and their control may increase the treatment efficacy and help to promote the use of RFA technique. PMID- 16253161 TI - [The role of hypoxia on expression of cyclooxygenase-2 in synovial fibroblasts derived from human temporomandular joint]. AB - OBJECTIVE: To investigate the effects of hypoxia on expression of COX-2 in synovial fibroblasts derived from human TMJ and to analyze the biological role of COX-2 on TMJ disorders. METHODS: Synovial fibroblasts were induced by hypoxia, COX-2 expression was determined by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting. Moreover, cells were stimulated by hypoxia or celecoxib or both of them, prostaglandin E(2) (PGE(2)), matrix metalloproteinase 2, 9 (MMP-2, MMP-9) release was detected by enzyme-linked immunosorbent assay (ELISA) or gelatin zymography. Cells that were placed under normoxic conditons were used as controls. RESULTS: (1) In response to hypoxia, at 4 h (1.51 +/- 0.34), 8 h (1.39 +/- 0.57), significant elevation of COX-2 mRNA expression occurred, compared with data (0.65 +/- 0.24), (0.71 +/- 0.15) under normoxic condition (both P < 0.05). (2) The elevation of COX-2 protein was detectable at 6 h (0.29 +/- 0.06), 12 h (0.51 +/- 0.09), 24 h (0.68 +/- 0.11), contrasted with normoxia (all P < 0.01). Therefore, it is suggested that COX-2 expression pattern present a time-dependent response to hypoxic conditions. (3) The concentration of PGE(2) released by synovial fibroblasts under 12 h hypoxic condition (7.6 ng/ml +/- 0.8 ng/ml) was significantly higher than those under normoxic condition (2.5 ng/ml +/- 0.4 ng/ml, P < 0.01) or in response to celecoxib combined with hypoxia (4.3 ng/ml +/- 0.4 ng/ml, P < 0.05). (4) The amount of MMP-2, MMP-9 produced by synovial fibroblasts under 12 h hypoxic condition was upregulated compared with that under normoxia or in response to celecoxib combined with hypoxia. CONCLUSIONS: Hypoxia is, at least in part, responsible for the pathogenesis of TMJ disorders, probably by promoting COX-2 expression of synovial fibroblasts derived from TMJ. PMID- 16253162 TI - [Regional modulation of primary motor cortex after peripheral nerve injury: a functional magnetic resonance imaging study]. AB - OBJECTIVE: To map dynamic changes of primary motor cortex after total brachial plexus traction injury by using functional magnetic resonance imaging, and to explore underlying probable mechanisms. METHODS: Five patients with total traumatic root avulsions of the brachial plexus underwent varied kinds of nerve transfer to restore partially shoulder or elbow function. Four of them (cases 1, 3, 4, 5) accepted the first fMRI examination prior to surgery treatment, and four of them (cases 2, 3, 4, 5) accepted second or third or fourth fMRI follow-up re examinations after surgery treatment. Maps of neuronal activation within the motor cortex were generated for both hands in each patient by using BOLD-fMRI and the cluster size and position were recorded. The motor tasks consisted of simple hand grasping of both hands respectively. Patients with paralytic hand were asked to complete task under "virtual" condition. The cluster size and intensity as well as location of motor activation within the primary motor cortex of the affected hand generated were compared with those of unaffected hand generated as reference in single subject, and the resultant maps of follow-up re-examinations were also compared with those of the prior examinations. RESULTS: All patients' unaffected hand movement generated strong signal change within the contralateral primary motor cortex. In contrast, the clusters generated by affected hand showed very small and lower intensity than usual (2 cases) or could not be induced (2 case) in the first examination that prior to surgery treatment and seemed larger gradually in the following re-examination with time increasing. CONCLUSION: Peripheral nerve injury can produced significant changes in the motor cortex of human brain. fMRI is a valuable tool to evaluate neural plasticity in motor cortex after peripheral nerve injury. PMID- 16253163 TI - [Application of oligonucleotide array in single nucleotide polymorphism typing of cytokines and cytokine receptors]. AB - OBJECTIVE: To develop an oligonucleotide array for single nucleotide polymorphism (SNP) typing of cytokines, such as tumor necrosis factor (TNF)-a, interleukin (IL)-10, tumor growth factor (TGF)-betal, IL-4, and IL-6, and their receptors and evaluate its function by direct sequencing. METHODS: According to relevant literature, SNP database of NCBI and SNP500 Cancer database of NCI, SNP loci and sequences of cytokines of clinical importance, TNF-a, IL-10, TGF-bl, IL-4 and IL 6, and matched cytokine receptors were chosen and 59 synthesized oligonucleotide probes were immobilized on a glass support, then the primer and Cy5-dCTP were used in multi-PCR, thus the products were labeled with Cy5. The labeled PCR products were hybridized with the probes in the array, and the signals were scanned by Scanner and then analyzed by Image software. Peripheral blood samples were collected from 80 healthy donors and 50 patients with uremia to isolate lymphocytes and DNA so as to undergo typing by this array, and the results were validated with direct sequencing. RESULTS: All the samples with PCR products except for 10 from uremia patients had been genotyped by cytokine array successfully. No diversity was found in genotyping for three times. Incorrect locus was not found with direct sequencing. CONCLUSION: With high specificity, sensitivity, repetitiveness, and throughout, and easy to manipulate, oligonucleotide array technique is an ideal molecular method for SNP genotyping of cytokine and cytokine receptor. PMID- 16253165 TI - [The molecular mechanism of PML in inducing apoptosis of bladder cancer cell line]. AB - OBJECTIVE: Study the affection of inducing expression of PML in the apoptosis and the molecular mechanism of bladder cancer. METHODS: PMEP4/PML inducible expression vector was transfected into bladder cancer UM-UC-2 cells by lipofectamine 2000 system. The positive clone cells were selected by 300 microg/ml hygromycin B and confirmed by laser confocal imaging system. Then, using the in vitro DNA ladder apoptotic assay and Western blot, the affection of inducing expression of PML on apoptosis and its molecular mechanism of bladder cancer cell was studied. RESULTS: Comparing with the vector control group, PML specific nuclear speckle significantly increased in the PMEP4/PML bladder cells. DNA ladder assay demonstrated bladder cancer cell expressing PML occurred apoptosis while the control vector cells were not influenced. Overexpression of PML could reduce Survivin expression and upregulate caspase3 and cleaved PARP protein expression CONCLUSION: Our results demonstrated that overexpression of PML could induce bladder cancer cell apoptosis through the caspase dependent pathways. PMID- 16253164 TI - [Effect of new photosensitizer CDHS801-mediated photodynamic therapy on bladder cancer: an experimental study]. AB - OBJECTIVE: To investigate the effects of dynamic photodynamic therapy (PDT) on bladder cancer. METHODS: Human bladder cancer cells of the line T24 were co cultured with CDHS801, a photosensitizer, and MitoTracker RED CMXRose and MitoTracker GREEN FM, mitochondria specific fluorescence probe dyes. Laser scanning confocal fluorescence microimaging system was applied to collect the fluorescence of the photosensitizer and the probes. T24 cells were cultured and divided into 4 groups: Group 1 as blank control group, Group 2 undergoing laser irradiation, Group 3, added with CDHS801 for 6h, and Group 4 (PDT group), added with CDHS801 and undergoing laser irradiation. The survival of the cells was examined by MTT colorimetric assay. The morphological changes and apoptosis of the photo-activated T24 cells were investigated by transmission electron microscopy, confocal laser scan microscopy, and flow cytometry. RESULTS: The fluorescence of the photosensitizer and that of the probes were detected in the cytoplasm and in the peri-nuclear region, mainly in the mitochondria, of the T24 cells. 2. The inhibitory rates of PDT on T24 cells were 0%, 7.3%, 10.8%, and 71.4% in the control group, Group 1, Group 2, and Group 3 respectively. T24 cell photo-activated with CDHS801 showed cell size shrinkage, condensed chromatin and formation of apoptotic body. Flow cytometry showed that apoptosis was seen in 55.31% of the photo-activated cells and peaked in the sub-G1 phase. However, no such changes were seen in the control group. CONCLUSION: CDHS801-based PDT can kill bladder cancer T24 cells. CDHS801 is localized in the cytoplasm and peri nuclear region, mainly mitochondria, of tumor cell. CDHS801 based PDT maybe eliminate the T24 cell by the induction of apoptosis. PMID- 16253167 TI - [Expression of amyloid beta protein and amyloid precursor protein after focal brain ischemia in human]. AB - OBJECTIVE: To observe the expression of amyloid beta precursor protein and amyloid beta protein in neurons of hippocampal CA1 region after brain ischemia in human. METHODS: Neuronal damage was examined by using HE staining, and expression of APP and Abeta(1-40) was determined by immunohistochemistry in the hippocampal CA1 region of the brain specimens of 43 patients who died 2 h-6 h, 7 h-24 h, 25 h 48 h, 49 h-72 h, 73 h-96 h, 97 h-144 h, or 145 h-168 h after cerebral ischemia and in 2 specimens of patients who died of other diseases as control group. RESULTS: The expression of Abeta(1-40) was 25.07 +/- 2.79 in the specimens 2 h-6 h after cerebral ischemia, peaked 73 h-96 h after cerebral ischemia (33.22 +/- 2.67), then decreased till 145 h-168 h after cerebral ischemia, however, all higher than that in the control group (2.88 +/- 0.18, all P < 0.05). The expression of beta-APP was 33.30 +/- 0.42 2 h-6 h after cerebral ischemia, was 28.11 +/- 2.03 7 h-24 h after cerebral ischemia, increased to peak value (32.32 +/- 1.36) 73 h-96 h after cerebral ischemia, and then decreased to 28.48 +/- 2.01, all higher than that of the control group (25.90 +/- 1.55) with significant differences between those 2 h-6 h and 73 h-96 h after ischemia and that of the control group (both P < 0.01). The increase of beta-APP was positively correlated with the expression of Abeta(1-40) 24 h after ischemia. CONCLUSION: The expression of beta-APP and that of Abeta(1-40) are up-regulated after cerebral ischemia, thus aggravating cerebral ischemia. PMID- 16253168 TI - [Application of Model for end-stage liver disease to predict the prognosis of and curative effect for chronic severe hepatitis]. AB - OBJECTIVE: To compare the capability in predicting the prognosis of chronic severe hepatitis among 3 prediction systems: Model for end-stage liver disease (MELD), Child-Turcotte-Pugh (CTP) system, and King's College Hospital (KCP) system, and to explore the capability of MELD in predicting the curative effect of artificial liver. METHODS: Scoring was made among 66 patients of chronic severe liver diseases with the underlying disease of hepatitis B, 11 in early stage, 14 in middle stage, 15 in late stage, and 26 unclassified, by MELD, CTP, and KCP systems. The accuracy of each system was evaluated by ROC curve, the differences between the systems was analyzed by Kaplan-Meier survival curve. RESULTS: The MELD score of the patients at admission predicted the mortality within 3 months with the c-statistic of 0.894, higher than those of the CTP and KCP systems (0.703 and 0.89 respectively). The MELD scores of the patients in the early stage was 24 +/- 4, significantly lower than those in the middle and late stages (31.11 +/- 2.90 and 41.38 +/- 9.98 respectively, all P < 0.01). The MELD score was positively correlated with the stage of disease (r = 0.737, P < 0.01). The mortally was 10.7% for the patients with an admission MELD score < or = 30, was 47% for the patients with an admission MELD score of 31 approximately 39, and was 60% the patients with an admission MELD score of > or = 40. CONCLUSION: The predictive capability of MELD system is better than the KCP and CTP systems. Artificial liver support treatment is the best choice e for the patients with an admission MELD score of 31-39. An admission MELD score > 40 is the indication for liver transplantation. PMID- 16253169 TI - [Late course conformal radiotherapy combined with chemotherapy for stage III and IV a nasopharyngeal carcinoma]. AB - OBJECTIVE: To evaluate the effect and acute toxicity of late course conformal radiotherapy combined with chemotherapy for stage III and IV a nasopharyngeal carcinoma (NPC). METHODS: Ninety-six patients with stage III and IV a NPC were randomly divided into 2 groups: test group (n = 46, undergoing late course conformal radiotherapy combined with chemotherapy) and control group (n = 50, undergoing conventional radiotherapy). Both groups were treated with one-period chemotherapy, including cisplantin, 5-fluouracil, and calcium folinate, before and after the radiotherapy. The radiotherapy of the test group consisted of 2 phases: 36.0 approximately 40.0 Gy in 18 approximately 20 fractions over 3.5-4 weeks as the first phase using conventional technique was delivered with 2 lateral opposing faciocervical fields, and then 30.0-46.0 Gy in 15-23 fractions over 3-4.5 weeks as the second phase using three-dimensional conformal radiotherapy (3D-CRT). RESULTS: The rates of complete remission (CR), partial remission (PR), and no change (NC) of the test group were 95.65%, 4.35%, and 0, not significantly different from those in the control group (90.00%, 10.00%, and 0, P = 0.287). The 1-year survival rate of the test group was 100%, not significantly different from that of the control group (96.00%, P = 0.170). The nasopharyngeal 1 year control rate of the test group was 97.83%, significantly higher than that of the control group (78.00%, P = 0.03). The distant metastasis rate of the test group was 8.70%, not significantly different from that of the control group (12.00%, P = 0.596). The incidence rates of radiological caries and irradiation-induced otitis media in the test group was 4.25% and 17.39% respectively, both significantly lower than those in the control group (26.00% and 42.00% respectively, P = 0.004 and P = 0.000). There was no significant difference in the incidence of nausea, vomiting, leukopenia, and severity of acute mucositis between these 2 groups. CONCLUSION: Late course conformal radiotherapy combined with chemotherapy effectively improves the disease control, delays the distant metastasis, and alleviates radioactivity damnification. PMID- 16253180 TI - [To understand male climacteric syndrome (andropause) once again]. PMID- 16253182 TI - [Therapeutic effectiveness of liver transplantation: a single center study of 203 consecutive cases]. AB - OBJECTIVE: To investigate the measures to further improve the therapeutic efficacy of liver transplantation. METHODS: The clinical data of 203 consecutive cases of orthotopic liver transplantation (OLT) performed in Zhongshan Hospital, Fudan University April 2001 to October 2004, with the indications for considering OLT of hepatocellular carcinoma (HCC) (142 cases), liver cirrhosis (36 cases), fulminant hepatic failure (7 cases), Wilson's disease (6 cases), and other end stage liver diseases (12 cases), including 199 cases of cadaveric OLT, inclusive of 2 cases of reduced-size and 1 case of splitting liver transplantation, and 4 cases of living-donor liver transplantation, were retrospectively analyzed. Multivariate analysis using Cox proportional hazards regression model was applied to determine the risk factors predicting liver transplantation prognosis for HCC. RESULTS: For the whole group of the 201 patients, the 1-year and 2-year cumulative survival rates were 85.0% and 82.4% respectively and the rejection rate was 12.3%. In the HCC group the 1- and 2-year cumulative survival rates were 80.2% and 78.4%, and the 1- and 2-year disease-free survival (DFS) rates were 85.3% and 80.3% respectively. HCC recurrence was observed in 20 patients after OLT with a recurrence rate of 14.1%. Multivariate analysis revealed that the tumor size and portal vein tumor thrombus were the most independent and statistically significant factors affecting the DFS. CONCLUSION: OLT may be the most effective treatment option for patients with end-stage liver diseases and may also provide the opportunity of curative treatment or survival improvement for selected patients with hepatic malignancies. PMID- 16253184 TI - [The influence of induced hypertension in acute phase on focal cerebral ischemia injury]. AB - OBJECTIVE: To study the influence of induced hypertension in acute phase on the infarct volume and regional cerebral blood flow (rCBF) during focal cerebral ischemia in rats. METHODS: The adult rat's models of permanent focal cerebral ischemia were established by permanent middle artery occlusion. Thirty-six rats were divided six groups. The rCBF of cortical penumbra during focal cerebral ischemia was determined by Laser-Doppler Flowmetry. After experiment was over, the brain was TTC stained and the infarct volume was calculated by computer image analytic system. RESULTS: The rCBF was resumed by (81.8 +/- 3.1)%, (56.0 +/- 2.1) and (38.8 +/- 2.0)% in induced hypertension groups compared with the basic rCBF. The volume of infarction was (14 +/- 7) mm(3), (90 +/- 24) mm(3) separately in group induced hypertension after 2 and 3 hour ischemia. Induced hypertension during 3 hours could reduce infarct volume significantly. CONCLUSION: To improve rCBF in penumbra may be one of protective mechanism of induced hypertension on brain ischemia. PMID- 16253183 TI - [The characteristics of gene mutations in Chinese patients with Charcot-Marie Tooth disease]. AB - OBJECTIVE: To study the characteristics of gene mutations in Chinese patients with Charcot-Marie-Tooth disease (CMT). METHODS: Real-time quantitative PCR, PCR SSCP, and/or direct sequencing were used to analyze the mutation of the pathogenic genes PMP22, MPZ, CX32, EGR2, GDAP1, NEFL, HSP22 and HSP27 in 113 probands of CMT families, 45 of which had family history, from different provinces in China. The whole family members of the subjects with abnormal electrophoretic bands and 50 normal controls underwent the same examination. RESULTS: Thirty-six cases of PMP22 duplication, 7 cases of CX32 mutation, 1 case of HSP22 mutation, 1 case of HSP27 mutation, 1 case of MPZ mutation, and 1 case of GDAP1 mutation were found in the 113 CMT probands. No point mutation was found in PMP22, EGR2 and NEFL genes. CONCLUSION: Among the Chinese CMT patients 31.9% are caused by PMP22 duplication, 6.2% by CX32, and 0.9% by HSP22, HSP27, MPZ and GDAP1. Point mutations of PMP22, EGR2 and NEFL are rare. PMID- 16253185 TI - [Feasibility of the programme of focusing on early childhood development in impoverished rural areas in China]. AB - OBJECTIVE: To study the feasibility of the programme "Focusing on early childhood development" in impoverished rural areas in China. METHODS: A questionnaire survey was conducted among 100 mothers, about half with junior middle school education, 18% with grade-school education, and 29% being illiterate, of children aged under two years in 7 villages in Changfeng County, Anhui Province, a native impoverished county, to understand their knowledge and attitudes about child healthcare. Then these mothers were randomly divided into 2 equal groups: experimental group, receiving face-to-face guidance in the practice of early healthcare for children at home with a pamphlet developed by WHO as teaching material; and control group without receiving guidance. Two months later the mothers received the second session of guidance. Six months after the same questionnaire survey was conducted among the 100 mothers to investigate the changes in knowledge and practice and Gesell development test was made to observe the changes in mental development. Two-days' training on the technique of early child healthcare in the form of reading, videotape presenting, and exercise, was given to 29 village doctors. Questionnaire survey was conducted among them before and 1 month after the training to understand the changes of their knowledge and attitudes towards early child healthcare. RESULTS: After the guidance the knowledge and fostering skills in terms of early child healthcare were improved significantly among the mothers of the experimental group in comparison with the mothers of the control group (P < 0.01). After the training most of the village doctors managed put the knowledge they had learned into practice, however, the percentage of those who lacked confidence in the smooth implementation of the programme remained unchanged (38% vs. 41%). Before the experiment the abilities in different fields were not significantly different between the children in these 2 groups. Six months after the implementation of the programme the abilities in fine motion, speech, and interpersonal communication were improved significantly among the children of the experimental group (all P < 0.05). CONCLUSION: Economic, simple, and effective, the programme "Focusing on early childhood development" is suitable for the impoverished rural areas in China. PMID- 16253186 TI - [Effects of aldose reductase on the transforming growth factor-beta1-induced expression of fibronectin and collagen IV: experiment with cultured rat mesangial cells]. AB - OBJECTIVE: To study the effects of aldose reductase (AR) on the transforming growth factor (TGF)-beta1-induced expression of fibronectin (FN) and collagen IV. METHODS: Restriction endonucleases digestion and ligation were used to reconstruct the eukaryotic expression plasmid pCDNA3-AR. Rat mesangial cells (MsCs) were isolated, cultured, and transfected with pCDNA3-AR, or blank vector. The AR expression in the MsCs was examined by immunofluorescence analysis. RT-PCR was performed to detect the mRNA expression of AR in the MsCs and Western blotting was used to detect the protein expression of AR. AR inhibitors (ARIs), Sorbinil and Zopolrestat were added and co-incubated, then TGF)-beta1 was added and Western blotting was used to analyze the protein expression of FN, collagen IV (Col IV), and mitogen-activated protein kinases (MAPKs) in the MsCs. RESULTS: Immunofluorescence analysis showed stronger expression of AR in the MsCs transfected with AR then in the normal MsCs and the MsCs transfected with blank vector. In comparison with the normal MsCs and those transfected with blank vector, the MsCs transfected with AR showed stronger protein expression of FN and Col IV (all P < 0.05). After incubation of ARIs the protein expression of FN and Col IV decreased by 1.8 and 2.0 times respectively in the MsCs transfected with AR (all P < 0.05). After stimulation of TGF-beta1, the protein expression of FN and Col IV increased in both the normal MsCs and those transfected with AR (P < 0.05 for the latter). After preincubation with ARIs the protein expression of FN and Col IV in the MsCs transfected decreased significantly (both P < 0.05). After stimulation of TGF-beta1, the normal MsCs showed increased expression of phospho ERK, phospho-JNK, and phospho-p38; the MsCs preincubated with ARIs showed reduced expression of phospho-JNK and phospho-p38; and the MsCs transfected with AR showed increased expression of phosphor-JNK (P < 0.05). CONCLUSIONS: AR can regulates the expression of FN and Col IV with the stimulation of TGF-beta1 as AR gene is one of the responsive genes of TGF-beta1, which may have relations with the activation of JNK-MAPK and p38-MAPK signaling pathways induced by TGF-beta1. AR may play a role in the pathogenesis of glomerulosclerosis. PMID- 16253188 TI - [Effects of modified acellularization process on porcine endogenous retroviruses in porcine aorta valves]. AB - OBJECTIVE: to study the effect of modified acellularization process on porcine endogenous retrovirus (PERV) in porcine aorta valves (PAVs). METHODS: Twenty aortic valves of pig were put into 0.1% trypsin solution, hypotonic and hypertonic TritonX-100, DNAse solution, RNAse solution, and Hanks solution in succession so as to remove the cells. The specimens of PAV were to undergo gross observation and microscopy before and after the acellularization procedure. Fracture test was made. Primers specific for the conservative gag gene of PERV were designed PCR and RT-PCR were used to detect the expression of gag. In addition, 20 samples of native PAV were collected. Peripheral mononuclear cells (PBMCs). Were isolated from 20 samples of porcine peripheral blood. Ten dogs underwent acellularized PAV replacement; 3 months later, samples of the dogs' peripheral blood were collected. Porcine kidney cells of the line PK15 were used as positive controls. RESULTS: Microscopy showed that all the cells were removed from the acellularized PAVs. Histological analysis showed that the major structural components were maintained. There was no significant difference in fracture strength between the native and acellularized PAVs (P > 0.05). PCR and RT-PCR showed a PERV 219 bp DNA fragment, 90%-95% homologous with the published PERV gene, in the genomic DNA of all native PAVs, pig PBMCs, and PK15 cells, but not in the acellularized PAVs and dog PBMCs. CONCLUSION: PERV exists in all native PAVs. The modified acellularization process succeeds in removing all the cell component and PERV in the PAVs, thus preventing cross-species transmission of PERV. PMID- 16253189 TI - [Angiotensin II and aldosterone stimulate alpha1-(I) procollagen mRNA expression in hepatic stellate cells via activation of ERK1/2 and AP-1]. AB - OBJECTIVE: To investigate the signal transduction mechanism underlying the effects of angiotensin II (Ang II) and aldosterone (Aldo) on the signal passageway of active protein-1 (AP-1). METHODS: In vitro, Hepatic stellate cells (HSCs) of the line HSC-T6 were cultured and treated with Ang II or Aldo, the principal effector molecules of the renin-angiotensin-aldosterone system (RAAS) for 10, 30, 60, 120, and 180 minutes respectively. The protein expression of phospho-P42/44 was detected by Western blotting. In addition, HSC-T6 cells were preincubated for 60 min with U0126, an inhibitor of MAPK/ERK kinase, irbesartan, an AT-1 receptor blocker, N-acetylcysteine (NAC), antioxidant, angiotensin converting enzyme inhibitor (ACEI), or tumor necrosis factor alpha (TNFalpha) prior to exposure to Ang II or Aldo. Then the protein expression of phospho P42/44 was measured by Western blotting. The DNA biding activity of AP-1 was analyzed by electrophoretic gel mobility shift assay (EMSA). By means of RT-PCR, the mRNA expression of alpha1 (I) procollagen was detected. RESULTS: The levels of phopho-ERK1/2 protein increased after the treatment of Ang II and Aldo at all time points and both peaked 10 minutes after (both P < 0.01). The levels of phopho-ERK1/2 protein of the irbesartan + Ang II and U0126 + Ang II groups were significantly lower than that of the Ang II group (both P < 0.01). The level of phopho-ERK1/2 protein of the Ang II group was lower than that of the TNFalpha group, however, was especially significantly lower than that of the Ang II + TNFalpha group (P < 0.01). The level of phopho-ERK1/2 protein of the U0126 + Aldo group was significantly lower than that of the Aldo group (P < 0.01). The phopho ERK1/2 protein level of the NAC + Aldo group was not significantly different from that of the Aldo group (P > 0.05). The phopho-ERK1/2 protein level of the Aldo group was lower than that of the TNFalpha group, however, was especially significantly lower than that of the Aldo + TNFalpha group (P < 0.01). The AP-1 DNA binding protein increased after the treatment of Ang II and peaked 30 min after. U0126, irbesartan, and NAC, as well as ACEUI, significantly inhibited the increased AP-1 DNA binding activity induced by Ang II. The AP-1 DNA binding protein increased after the treatment of Aldo and peaked twice, 30 min and 240 min after. U0126 and NAC significantly and NAC partly inhibited the increased AP 1 DNA binding activity induced by Aldo. CONCLUSION: Stimulation of HSC by Ang II and Aldo results in activation of AP-1 via ERK1/2 pathway leading to up regulation of AP-1 target gene alpha1 (I) procollagen mRNA expression. PMID- 16253190 TI - [Effects of rhein on activity of caspase-3 in kidney and cell apoptosis on the progression of renal injury in glomerulosclerosis]. AB - OBJECTIVE: To investigate the effects of rhein on the progression of renal injury and cell apoptosis in glomerulosclerosis, and further explore the protective mechanism of rhein on glomerulosclerosis. METHODS: Glomerulosclerosis models were made for SD rats by unilateral nephrectomy and being injected with Adriamycin into caudal vein, and randomly divided into control group, renal disease group, Rhein treatment group and Benazepril treatment group, and 6 rats in each group were killed at the 6th, 8th, 10th, 12th week respectively. The apoptosis protease 3 (caspase-3) in renal cortex was determined by immunohistochemistry stain method, and the activity of caspase-3 was measured by colorimetry, and the activity of nuclear factor-kappa B (NF-kappaB) was analyzed by gel electrophoretic mobility shift assay (EMSA), and renal tissue cell apoptosis was tested by terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL) in order to observe expressions of caspase-3 and NF-kappaB and cell apoptosis of renal tissue. RESULTS: Renal disease group presented with distinct proteinuria, decreasing of blood albumin content and increasing of cholesterol concentration. Glomerulosclerosis index, apoptosis index, activity of NF-kappaB and expression of caspase-3 in renal disease group were more significantly higher than those in control group (P < 0.05 or P < 0.01) as time passed. Compared with the other time points in renal disease group, there were a great number of TUNEL positive cells observed at the 10th week, slightly higher than that at the 12th week (9.3 +/- 2.3 vs 8.4 +/- 1.2, P > 0.05), the expression of Caspase-3 was also most obvious at the 10th week, significantly higher than that at the 12th week (11.4 +/- 2.5 vs 8.2 +/- 1.7, P < 0.05), which mainly located around capillary vessel in renal cortex, tending to be consistent with apoptosis cells expression. After the 8 weeks treatment of rhein or Benazepril, the number of TUNEL-positive cells significantly decreased and maintained at a certain level, and the activity of NF-kappaB and expression of caspase-3 decreased (P < 0.05), and renal pathological changes and biochemical changes improved magnificently, moreover, the expression of caspase-3 showed positive correlation with apoptosis index (r = 0.836, P < 0.01). CONCLUSION: Rhein could have significant protective effects on the progression of renal injury, and might regulate pathological changes by influencing the activities of NF-kappaB and caspase-3 in the early phase of glomerulosclerosis. Therefore, down-regulating caspase-3 expression in kidney might be one of the molecular mechanisms in the way that rhein could alleviate renal tissue cell apoptosis in glomerulosclerosis. PMID- 16253191 TI - [Expression of cancer-related indices in different types of cystitis glandularis and clinical significance thereof]. AB - OBJECTIVE: To explore the rationality of clinical typing of cystitis glandularis (CG) and the potential of cancerization of different types of CG. METHODS: Flow cytometry was performed to examine the ploidy of 25 fresh bladder specimens of patients with CG resected during operation, 13 high risk type and 12 low risk type, and 7 specimens of normal mucosa of bladder, by calculating the DNA index (DI). Immunohistochemistry was used to detect the expression of proliferating cell nuclear antigen (PCNA), mutant p53, p21ras, Rb, and bcl-2 in other 38 preserved specimens of CG resected during operation, 18 high risk type and 20 low risk type, and 5 specimens of normal bladder. RESULTS: The DI was 1.00 +/- 0.03 in the normal bladder group, 1.05 +/- 0.07 in the high risk type CG group, and 1.01 +/- 0.05 in the low risk type CG, without significant differences between any 2 groups (t = -1.639, P = 0.115). The expression of PCNA and expression of p53 were negative in the low risk group, and PCNA was expressed in 5 specimens of high risk type CG (27.8%), and not expressed in the low risk type CG (P = 0.017). p53 was expressed in 4 specimens of high risk type CG (22.2%), and not expressed in the low risk type CG (P = 0.041). There were no significant differences in the expression of p21, Rb, and bcl-2 between the high and low risk groups. CONCLUSION: High risk type and low risk type of CG are both benign lesions. High risk type CG may be more likely to cancerate. It is reasonable to distinguish different types of CG. p53 gene may play an important role in the canceration of CG. PMID- 16253192 TI - [Prenatal diagnosis of X-linked anhidrotic ectodermal dysplasia with X-chromosome inversion]. AB - OBJECTIVE: To investigate the possibility of prenatal diagnosis of the fetal suspected to be affected by anhidrotic ectodermal dysplasia (EDA) in a family with X-linked EDA so as to provide a basis for prenatal diagnosis and genetic counseling of this disorder. METHODS: Pedigree analysis and genetic counseling were performed in a family after a proband was diagnosed with EDA. The peripheral blood samples were collected from the proband, a 12-year-old boy, his mother, and his 2 aunts, one being pregnant, to undergo chromosome karyotype analysis. The fetus Puncture of umbilical vein was performed to collect the blood of fetus for chromosome examination. Induced abortion was conducted due to the diagnosis of the fetus with EDA. Autopsy, immunohistochemistry of the skin tissues of face, breast, epigastrium, and thigh, and X-ray photography of the lower jawbone were made. RESULTS: Pericentric inversion occurring at one of the X-chromosome [inv (x) (p22q13)] was found in the proband and his nephew (the fetus), both patients, and his mother and his second aunt (the pregnant woman), both carriers. Autopsy of the fetus showed epidermis dysplasia and deficiency of hair follicle and sebaceous gland. Immunohistochemistry showed that epithelial membrane antigen and cytokeratin were negatively expressed in the fetal skin tissues. CONCLUSION: Pedigree analysis and genetic counseling for the family members of EDA patients and prenatal and postpartum examination for the fetus help diagnose EDA. PMID- 16253193 TI - [Prosthetic valve replacement in pediatric patients: analysis of 105 cases]. AB - OBJECTIVE: To summarize the experience in prosthetic valve replacement in pediatric patients. METHODS: The clinical data of consecutive 105 children, 63 male and 42 female, aged 13 +/- 3 (1.5-16), with the underlying diseases of rheumatic heart disease (n = 55), congenital heart diseases (n = 43), and infective endocarditis (n = 5), with the preoperative cardiac function of class II (n = 36), or class III or IV (n = 69), who underwent prosthetic valve replacement, including replacement of mitral valve (n = 58), aortic valve (n = 28), tricuspid valve (n = 5), mitral and aortic valves (n = 13), and mitral and tricuspid valves (n = 1), and tricuspid valvuloplasty (n = 26), from May 1984 to May 2004, were respectively analyzed. Mechanical valves were used and took low dose anticoagulant warfarin was administrated to all cases. Postoperatively prothrombin time (PT) and international normal ratio (INR) were observed. Follow up lasting 10 +/- 4 years (6 months to 20 years) was performed among 100 discharged patients with a follow-up rate of 91.0%. RESULTS: There were 5 early deaths (4.8%), mostly owing to serious low cardiac output syndrome, and 4 late deaths owing to endocarditis (n = 2), heart failure (n = 1), and serious arrhythmia (n = 1). The 87 surviving patients showed their cardiac function of class I-III. No serious complications related to anticoagulation and prosthesis dysfunction had been found. CONCLUSION: Prosthetic valve replacement with mechanical valve can be performed in pediatric patients with good outcome. Larger type of mechanical valve should be used as possible. Additional surgical procedures may be required to correct the concomitant malformations during the operation. Postoperative long-term low dose anticoagulant treatment is safe. PMID- 16253203 TI - Morphology characterization of emulsions by differential scanning calorimetry. AB - This article is a review of some results obtained by Differential Scanning Calorimetry (DSC) for characterizing the morphology of emulsions. In a classical DSC experiment, an emulsion sample is submitted to a regular cooling and heating cycle between temperatures that include freezing and melting of the dispersed droplets. By using the thermograms found in the literature for various emulsions, how to get information about the solidification and melting, the presence of solute, the emulsion type, the transfer of matter, the stability and the droplet size is shown. PMID- 16253202 TI - Evaluation of PCR based coprodiagnosis of human opisthorchiasis. AB - In this study, a recently developed PCR test for the detection of Opisthorchis viverrini in human faecal samples was evaluated using two parasitological methods as references. During a survey of foodborne trematodes (FBT) in the Vientiane Province, Lao PDR, 85 samples were collected and evaluated for FBT eggs by the Kato Katz (KK) technique, the formalin ethyl acetate concentration technique (FECT) and a PCR analysis for the distinction between O. viverrini and other FBT. The two parasitological methods did not differ in the ability of detecting FBT eggs, and a single KK reading was characterized by a sensitivity of 85% when compared to two FECT readings. The PCR tested positive only in cases where eggs had been demonstrated by parasitological examination. However, the PCR tested negative in some samples with very high egg counts. Demonstrating a PCR sensitivity of approximately 50% in samples with faecal egg counts>1000, the previously reported PCR sensitivity based on in vitro studies was not supported. It is believed that technical problems rather than diagnostic reference related issues were responsible for the relatively low PCR performance. Further studies should aim at optimizing DNA extraction and amplification, and future PCR evaluation should include specificity control such as the scanning electron microscopy of eggs in test samples or the expulsion of adult trematodes from PCR tested individuals. PMID- 16253204 TI - Alveolar bone osteoblast differentiation and Runx2/Cbfa1 expression. AB - Alveolar bone cells have a unique origin and functionality, but may resemble skeletal osteoblasts. Osteoblast differentiation and gene expression are regulated by the Runx2/Cbfa1 transcription factor. However, most studies on Runx2/Cbfa1 expression have been on rodent cells and the few studies on human osteoblasts have had differing results. The purpose of this study was to characterize Runx2/Cbfa1 expression in primary cell cultures derived from human alveolar bone. An alveolar bone chip was incubated in alpha-minimum essential medium (alpha-MEM) supplemented with fetal calf serum (10% FCS). Explant cultures were harvested after 3-4 weeks of outgrowth and grown in alpha-MEM with FCS. This media was supplemented with ascorbate, beta-glycerophosphate and dexamethasone to promote osteoblast differentiation over 14 days. RT-PCR analysis and Western blots showed a rapid increase in Runx2/Cbfa1 mRNA (2.1-fold) and protein (2.3 fold) levels in 3 days, followed by a slight decline. There was also a rapid increase in bone sialoprotein expression (2.9-fold) in 3 days, followed by a further increase (3.6-fold) at 14 days. There was a slower increase in alkaline phosphatase expression (1.6-fold) and activity (3.1-fold) over 7 days, followed by a gradual decline. In contrast, collagen mRNA levels showed little change over 14 days. These findings attest to the osteogenic potential of primary cell cultures derived from human alveolar bone. Osteoblastic differentiation in human alveolar bone involves an increase in Runx2/Cbfa1 expression that may be an important component of the differentiation process. PMID- 16253205 TI - Using controlled comparisons in disgust psychopathology research: the case of disgust, hypochondriasis and health anxiety. AB - The present paper describes the results of a study investigating the relationship between measures of disgust and measures of hypochondriasis and health anxiety. The results indicated that (1) there were highly significant correlations between measures of trait disgust and disgust sensitivity and measures of hypochondriasis and health anxiety, (2) the relationship between disgust sensitivity measures and hypochondriasis and health anxiety were still significant even when levels of trait anxiety were controlled for, but (3) controlled comparisons revealed that the measures of disgust also predicted scores on measures of disgust-irrelevant control psychopathologies (claustrophobia and height phobia)--even after trait anxiety had been partialled out. In addition, the series of multiple regressions carried out clearly indicated that trait anxiety and disgust sensitivity appear to be independent constructs each of which have relationships with anxious psychopathologies over and above the effect of the other. The discussion explores the nature of the possible relationships between disgust, hypochondriasis and health anxiety, and also looks at the implications for disgust psychopathology research of using controlled comparisons which indicate the existence of significant relationships between measures of disgust and anxious psychopathologies that, a priori, would be considered to be disgust irrelevant. PMID- 16253206 TI - Treating disgust reactions in contamination-based obsessive-compulsive disorder. AB - Contamination fear associated with obsessive-compulsive disorder (OCD) has features that strongly suggest it is based in part on disgust. In particular, sympathetic magic, or the tendency for disgust-evoking stimuli to transfer that property to previously neutral stimuli, is common in contamination fear. Treatment for OCD typically involves exposure with response prevention for feared stimuli. Unexamined, however, has been the habituation for specific disgust stimuli among individuals with contamination fear. This study is a preliminary investigation with a group of primary contamination fearful participants diagnosed with OCD (C-OC; n = 9) compared to a group with primarily other symptoms of OCD (O-OC; n = 8). All participants were exposed to a set of disgust stimuli that were not associated with anxiety reactions for 30 min across five sessions, and exposed to anxiety-evoking stimuli specific to their OCD symptoms for 30 min. Comparisons showed that, while both groups had reduced disgust reactions, the C-OC group habituated more slowly and to a lesser degree than the O-OC group. No differences existed between the groups for anxiety reduction. The findings suggest that contamination fear is based in part on disgust reactions, and that disgust is amenable to exposure-based interventions. PMID- 16253207 TI - The conserved C-termini contribute to the properties of spider silk fibroins. AB - Spider silk fibroins can adopt different structural states at high protein concentrations. They are soluble within the spinning dope of the glands, but readily converted into insoluble polymers upon extrusion. A contribution of the C termini to the maintenance and conversion of these states is suggested by their predicted secondary structures and biochemical behavior in vitro. Special sequence parts endow the C-termini with the capability to promote both the solubility and aggregation of the fibroins depending on the environmental conditions. PMID- 16253208 TI - Cdh1-APC/C, cyclin B-Cdc2, and Alzheimer's disease pathology. AB - The anaphase-promoting complex/cyclosome (APC/C) is a key E3 ubiquitin ligase complex that functions in regulating cell cycle transitions in proliferating cells and has, as revealed recently, novel roles in postmitotic neurons. Regulated by its activator Cdh1 (or Hct1), whose level is high in postmitotic neurons, APC/C seems to have multiple functions at different cellular locations, modulating diverse processes such as synaptic development and axonal growth. These processes do not, however, appear to be directly connected to cell cycle regulation. It is now shown that Cdh1-APC/C activity may also have a basic role in suppressing cyclin B levels, thus preventing terminally differentiated neurons from aberrantly re-entering the cell cycle. The result of an aberrant cyclin B induced S-phase entry, at least for some of these neurons, would be death via apoptosis. Cdh1 thus play an active role in maintaining the terminally differentiated, non-cycling state of postmitotic neurons--a function that could become impaired in Alzheimer's and other neurodegenerative diseases. PMID- 16253209 TI - Impairment of cardiomyogenesis in embryonic stem cells lacking scaffold protein JSAP1. AB - We previously reported that c-Jun NH(2)-terminal kinase (JNK)/stress-activated protein kinase-associated protein 1 (JSAP1), a scaffold protein for JNK signaling, is important in embryonic stem (ES) cells during neurogenesis. In that study, we also observed the altered expression of mesodermal marker genes, which indicated that JSAP1 is involved in the differentiation of mesodermal lineages. Here, we investigated the function of JSAP1 in cardiomyocyte development using JSAP1-null ES cells, and found that cardiomyogenesis was impaired in the JSAP1 null mutant. The JSAP1 deficiency resulted in lower gene expression of the cardiac transcription factor Nkx2.5 and contractile proteins. In contrast, the mutant showed a significantly higher expression of mesoderm-related markers other than those of the cardiomyocyte lineage. Together, these results suggest that JSAP1 may be important for the differentiation of the mesodermal lineages, functioning as a positive factor for cardiomyocyte differentiation, and as an inhibitory factor for differentiation into other lineages. PMID- 16253210 TI - NRH:quinone reductase 2: an enzyme of surprises and mysteries. AB - Quinone reductase 2 has been discovered in 1961 and rediscovered in 1997. Because of its sequence homology with quinone reductase 1, it has been suspected to detoxify quinones. Ten years later, evidences begin to point to a versatile role of this enzyme. Indeed, QR2 is strongly suspected to be the molecular target of anti-malarian drugs such as chloroquin or paraquine, and of red wine-derived resveratrol that might be responsible for the so-called French paradox. It also is identical to the melatonin binding site MT3, and might therefore be a rationale explanation for the antioxidant role of melatonin. Finally QR2 might be implicated in the toxicity, in vivo, of quinones such as menadione. The present commentary attempts to summarize this information and discusses a series of hypotheses. PMID- 16253211 TI - Role of elevated S-adenosylhomocysteine in rat hepatocyte apoptosis: protection by betaine. AB - Previous studies from our laboratory have shown that ethanol consumption results in an increase in hepatocellular S-adenosylhomocysteine levels. Because S adenosylhomocysteine is a potent inhibitor of methylation reactions, we propose that increased intracellular S-adenosylhomocysteine levels could be a major contributor to ethanol-induced pathologies. To test this hypothesis, hepatocytes isolated from rat livers were grown on collagen-coated plates in Williams' medium E containing 5% FCS and exposed to varying concentrations of adenosine in order to increase intracellular S-adenosylhomocysteine levels. We observed increases in caspase-3 activity following exposure to adenosine. This increase in caspase activity correlated with increases in intracellular S-adenosylhomocysteine levels and DNA hypoploidy. The adenosine-induced changes could be significantly attenuated by betaine administration. The mechanism of betaine action appeared to be via the methylation reaction catalyzed by betaine-homocysteine methyltransferase. To conclude, our results indicate that the elevation of S adenosylhomocysteine levels in the liver by ethanol is a major factor in altering methylation reactions and in increasing apoptosis in the liver. We conclude that ethanol-induced alteration in methionine metabolic pathways may play a crucial role in the pathologies associated with alcoholic liver injury and that betaine administration may have beneficial therapeutic effects. PMID- 16253212 TI - Oxidized LDL, a critical factor in atherogenesis. PMID- 16253213 TI - Questioning the relevance of circulating cardiac progenitor cells in cardiac regeneration. PMID- 16253214 TI - Integrins, membrane-type matrix metalloproteinases and ADAMs: potential implications for cardiac remodeling. AB - The concept of myocardial remodeling links an initial pathological insult to a progressive geometric change of the ventricle. Currently, our concepts of the remodeling process have evolved to include not only changes in ventricular size and shape, but cellular and molecular remodeling, particularly as the ventricle evolves towards failure. In recent years, much attention has focused on the role of cell-extracellular matrix (ECM) connections in this process. In this review, we will specifically delineate how cell membrane-linked molecules of three classes: integrins, membrane-type matrix metalloproteinases, and ADAMs (A Disintegrin And Metalloproteinase) might play crucial roles in myocardial remodeling. These molecules are essential for cell-ECM adhesion, cell signaling, matrix modification, and proteolysis of surface receptors. Our goal is to put forth concepts on how they might interrelate to modulate the remodeling process in the heart. PMID- 16253215 TI - Polyethylene glycosylation prolongs the circulatory stability of recombinant human butyrylcholinesterase. AB - Previous studies in rodents and non-human primates have demonstrated that pretreatment of animals with cholinesterases could provide significant protection against organophosphate (OP) nerve agent toxicity. Gene delivery/therapy is emerging as an approach to achieve high-level expression of proteins in vivo that are very similar to their native counterparts. Recently, adenoviral (Ad) vectors have proven to be excellent vehicles for delivering genes to cells in vitro and in vivo. In this study, we explored the use of the newly designed AdenoVATOR system for the expression of recombinant human butyrylcholinesterase (rHu BChE) in human embryonic kidney 293A (HEK-293A) cells. In these cells, rHu BChE was expressed as mostly tetrameric form by the simultaneous expression of proline rich attachment domain. By optimizing the culture conditions, 1.5-2.0 U/ml of rHu BChE could be expressed in HEK-293A cells. Recombinant Hu BChE was purified to homogeneity by ammonium sulfate fractionation followed by affinity column chromatography using procainamide Sepharose and cobalt Sepharose gels. The enzymatic and physico-chemical properties of purified rHu BChE were similar to those of native serum-derived Hu BChE. To determine the suitability of this preparation for use as an antidote against highly toxic nerve agents, its pharmacokinetics were evaluated in mice. Recombinant Hu BChE exhibited a mean residence time of 18.3 h which was 2.5-fold shorter than that observed for native Hu BChE in mice. However, rHu BChE chemically modified with polyethyleneglycol (PEG) displayed a mean residence time of 36.2 h suggesting that PEG-modification can prolong the circulatory stability of rHu BChE. The efficacy of Ad-Hu BChE to induce the production of therapeutic levels of bioscavenger in vivo is under evaluation. PMID- 16253216 TI - Determination of phase transition temperatures of lipids by light scattering. AB - Various techniques have been proposed to specify the phase transition temperatures of surfactant molecules. The work reported herein deals with a new general method of T(c) determination based on the optical properties' modifications of aqueous surfactant solutions when the phase transitions occur in the phospholipid membrane. The shape alteration of supramolecular systems induced by the phase transition was correlated with the refraction and absorption coefficients of their aqueous dispersion. The mean count rate (average number of photons detected per second) measured with a Zetasizer Nano-S model ZEN1600 Dynamic Light Scattering Instrument, is representative of an emerging macroscopic phenomenon, but not directly size dependent and has been adapted to our expectations. Changes in the measured scattering intensity reflect changes in the optical properties of the material during temperature variations. Thus, this method allowed to specify the phase transition temperature of many natural or synthetic surfactants independently of their polar head or hydrophobic part. PMID- 16253217 TI - Human hepatocytes secrete soluble CD14, a process not directly influenced by HBV and HCV infection. AB - BACKGROUND: Chronic hepatitis B (HBV) and hepatitis C (HCV) patients have elevated plasma levels of soluble CD14 (sCD14). We examined whether human hepatocytes produce sCD14 in vivo, and whether HBV or HCV infections influence this chimeric production. METHODS: uPA-SCID mice were transplanted with primary human hepatocytes and some animals were subsequently infected with HBV or HCV. Plasma from these mice was analyzed for the presence of human sCD14. The liver was examined via immunohistochemistry. RESULTS: A soluble form of human CD14 could be detected in the plasma from successfully transplanted mice, while it was completely absent in non-transplanted control animals. The isoform of this human sCD14 corresponded with the most abundant isoform found in human plasma. CD14 levels in circulation were not significantly different between non-infected, HBV infected and HCV infected animals. CONCLUSIONS: Our data indicate that human hepatocytes produce sCD14 in vivo, and that liver cells might be the major source of sCD14 in normal human plasma. In addition we demonstrate that HBV and HCV infections have no direct influence on the production of sCD14 by human hepatocytes in this chimeric model. PMID- 16253218 TI - Structural and functional analyses of mutations of the human phenylalanine hydroxylase gene. AB - BACKGROUND: Phenylketonuria (PKU) is an inborn error of metabolism that results from a deficiency of phenylalanine hydroxylase (PAH). We demonstrated PAH mutational spectrum from patients with PKU, including 10 novel and 3 tetrahydrobiopterin (BH(4))-responsive mutations. In this study, 11 PAH missense mutations, including 6 novel mutations (P69S, G103S, L293M, G332V, S391I, A447P) found in our previous study, 2 mutations common in east Asian patients with PKU (R243Q, R413P), and 3 tetrahydrobiopterin (BH(4))-responsive mutations (R53H, R241C, R408Q) have been functionally and structurally analyzed. METHODS: A transient protein overexpression system and an in vitro BH(4)-responsiveness study were used. The effects of PAH missense mutations on the PAH protein structure were also analyzed. To determine the conservation of 12 mutated residues, PAH was aligned using BLAST against full genomic sequences of 221 different species. Model structures of PAH protein and the composite tetramer were constructed using the software program, SHEBA. RESULTS: No PAH activity was detected for some mutants. However, the residual activities associated with other mutants ranged over a wide spectrum. The missense mutations responsive to BH(4) were not highly conserved throughout the 43 species in the multiple sequence alignment that encode PAH. The composite model structure of PAH revealed that dimer stability was reduced in the BH(4)-responsive mutants, whereas tetramer stability remained normal. CONCLUSION: This expression study analyzed PAH mutations and model structures of mutant PAH proteins are proposed. Correlation between the proposed mutant PAH structures and functions are suggested. PMID- 16253219 TI - Valid carbohydrate-deficient transferrin testing. PMID- 16253220 TI - Do platelet apoptosis, activation, aggregation, lipid peroxidation and platelet leukocyte aggregate formation occur simultaneously in hyperlipidemia? AB - OBJECTIVES: The circulating lipoproteins may cause some abnormalities in platelet composition and function in hypercholesterolemia. The aim of this study was to investigate whether platelet apoptosis, platelet activation, platelet aggregation, platelet-leukocyte aggregate (PLA) formation and lipid peroxidation occur simultaneously in hyperlipidemia. DESIGN AND METHODS: Expression of GpIIb/IIIa (CD41a), P-selectin (CD62-P), platelet-bound fibrinogen (antifibrinogen), platelet membrane phosphatidylserine (PS), platelet-monocyte aggregates (mono-PLA) and platelet-neutrophil aggregates (neut-PLA) was measured in eight hyperlipidemic and eight normal subjects using flow cytometry. ADP (10 microM) was used to activate platelets. Furthermore, ADP induced platelet aggregation responses, platelet malondialdehyde (MDA) and glutathione (GSH) levels were determined. RESULTS: Before platelet activation, platelet CD62-P, antifibrinogen, annexin-V, mono-PLA, neut-PLA and platelet MDA levels as well as platelet aggregation responses in the hyperlipidemics were significantly higher than those in the controls (P<0.01, P<0.01, P<0.01, P<0.001, P<0.001, P<0.01, P<0.001, respectively), whereas GpIIb/IIIa expression and GSH levels were not different significantly (P > 0.05). In the control group, CD62-P, antifibrinogen and annexin-V levels increased significantly after ADP activation (P<0.05, P<0.05, P<0.01, respectively). In hyperlipidemic subjects, annexin-V expression increased significantly after activation (P<0.01), whereas expression of GpIIb/IIIa, CD62-P and antifibrinogen remained unchanged (P>0.05). The levels of total cholesterol (T-CHO), low density lipoprotein cholesterol (LDL-C), serum fibrinogen (S-FGN) and high density lipoprotein cholesterol (HDL-C) in patients were found to be correlated with platelet CD62-P, antifibrinogen, annexin-V, mono PLA and MDA. CONCLUSIONS: In conclusion, it seems that in hyperlipidemia, some platelets are in an activated state in circulation, and that increased lipid peroxidation, early apoptosis, platelet-leukocytes aggregate formation and platelet aggregation altogether accompany this process. PMID- 16253221 TI - Building an allergens ontology and maintaining it using machine learning techniques. AB - Ontologies are widely used for formalizing and organizing the knowledge of a particular domain of interest. This facilitates knowledge sharing and re-use by both people and systems. Ontologies are becoming increasingly important in the biomedical domain since they enable knowledge sharing in a formal, homogeneous and unambiguous way. Knowledge in a rapidly growing field such as biomedicine is usually evolving and therefore an ontology maintenance process is required to keep ontological knowledge up-to-date. This work presents our methodology for building a formally defined ontology, maintaining it exploiting machine learning techniques and domain specific corpora, and evaluating it using a well-defined experimental setting. The application of this methodology in the allergen domain is then discussed in detail presenting the ontology built, the specific techniques used and the evaluation settings. PMID- 16253222 TI - Probabilistic prediction of protein-protein interactions from the protein sequences. AB - Prediction of protein-protein interactions is very important for several bioinformatics tasks though it is not a straightforward problem. In this paper, employing only protein sequence information, a framework is presented to predict protein-protein interactions using a probabilistic-based tree augmented nai ve (TAN) Bayesian network. Our framework also provides a confidence level for every predicted interaction, which is useful for further analysis by the biologists. The framework is applied to the yeast interaction datasets for predicting interactions and it is shown that our framework gives better performance than support vector machine (SVM). The framework is implemented as a webserver and is available for prediction. PMID- 16253223 TI - Distribution of aliphatic compounds in bivalve mollusks from Galicia after the Prestige oil spill: spatial and temporal trends. AB - The content and distribution of n-alkane (C8-C35) and isoprenoid (pristane and phytane) hydrocarbons were investigated in two species of bivalve mollusk, mussel (Mytilus galloprovincialis) and cockle (Cerastoderma edule), collected at different points of the Galicia littoral zone during the period from December 2002 to February 2003 (after the Prestige oil spill). Samples were analyzed by high-resolution gas chromatography equipped with a flame ionization detector. The highest levels were found in mussels and cockles coming from two estuarine bays, Rias de Arousa and Vigo. Hydrocarbons with carbon chain length > 30 were detected and determined in all samples. The abundance of these hydrocarbons in biota could be interpreted with regard to the feeding and living habits. Chemometric techniques have been employed to analyze data and determine the potential source of hydrocarbon contamination. Differences between mussels and cockles were observed in relation to aliphatic content. According to the data analysis, the main source of hydrocarbon contamination of investigated samples seems to be more related to the intense traffic of vessels in these estuarine bays than to the Prestige oil spill. PMID- 16253224 TI - Different effects of GABAergic receptors located in the ventral tegmental area on the expression of morphine-induced conditioned place preference in rat. AB - In the present study, an unbiased conditioned place preference paradigm was used to study the effects of intra-ventral tegmental area injections of Gama-amino butyric acid (GABA)-A and B (GABA(A) and GABA(B)) receptor agonists and antagonists on the expression of morphine-induced conditioned place preference (CPP) in rats. Subcutaneous (s.c.) injections of morphine sulfate (5 mg/kg) induced CPP. Intra-ventral tegmental area administration of the GABA(A) receptor agonist, muscimol (6 microg/rat) reduced the expression of morphine-induced CPP. Muscimol (25 microg/rat) increased the expression of CPP induced by morphine. A reduction of the expression of morphine-induced CPP was observed on intra-ventral tegmental area injection of GABA(A) receptor antagonist bicuculline (25 microg/rat). Bicuculline (10 microg/rat) increased the expression of CPP induced by morphine. Baclofen (12 microg/rat) increased where as (19 and 25 microg/rat) reduced the expression of morphine-induced CPP. Injection of CGP38345 (10, 19, 25 and 50 microg/rat) into the ventral tegmental area significantly reduced the expression of CPP induced by morphine. It is concluded that GABA(A) and GABA(B) receptor subtypes within the ventral tegmental area may have different effects on the expression of morphine-induced CPP. PMID- 16253225 TI - Role of GABAA alpha5-containing receptors in ethanol reward: the effects of targeted gene deletion, and a selective inverse agonist. AB - GABA(A) receptors containing alpha5 subunits have been suggested to mediate the rewarding effects of ethanol. We tested this hypothesis in mice with deletion of alpha5 subunits. alpha5 knockout mice did not differ from wildtypes in operant responding for 10% ethanol/10% sucrose, but responded less for 10% sucrose. The benzodiazepine (BZ) site inverse agonist, Ro 15-4513, has higher affinity for GABA(A) receptors containing 5 subunits and dose-dependently (0-27 mg/kg, i.p.) reduced lever pressing for ethanol/sucrose in wildtype mice, but had less effect in knockout mice; lever pressing for sucrose was unaffected. These data suggest that alpha5 subunits are not essential for ethanol reward, but the reduction of operant responding for ethanol by Ro 15-4513 is mediated by alpha5-containing GABA(A) receptors. In measures of ethanol consumption, alpha5 knockout mice did not differ from wildtypes at low ethanol concentrations (2-8%), but consumed less ethanol at higher concentrations; these differences were not attributable to increased behavioural disruption of the knockout by ethanol, since no differences were seen in sensitivity to ethanol's sedative or ataxic effects. Ro 15-4513's ability to reduce ethanol consumption was unaffected, suggesting that this effect is not mediated by the alpha5 subtype. Secondly, we tested the ability of a novel alpha5-efficacy-selective benzodiazepine receptor ligand, alpha5IA-II, that possesses greater inverse agonist activity at alpha5- than at alpha1-, a2- or alpha3-containing GABA(A) receptors, to influence operant responding. alpha5IA-II (0.03-3 mg/kg) dose-dependently decreased lever pressing for 10% ethanol, the minimally effective dose of 1 mg/kg, corresponding to over 90% receptor occupancy, but did not affect lever pressing for 4% sucrose. Although inverse agonists acting at alpha5-containing receptors reduce ethanol self administration, alpha5 subunits may not be essential to signaling ethanol reward. PMID- 16253227 TI - Individual differences in drug dependence in rats: the role of genetic factors and life events. AB - Drug dependence and addiction is a chronic mental illness that has far reaching consequences for society in terms of economic loss, health costs and judicial problems. A crucial question in drug addiction, is what factors are involved in its aetiology, and especially what mediates the shit from use to abuse. As in most other mental illnesses, addiction can best be described using the so-called three hit model, which states that a disease results from an interaction between genetic factors, early lie events and late environmental factors. However, the precise nature of these factors still remains to be elucidated. This present review discusses the results from an animal model in which these three different hit are currently being investigated. The apomorphine susceptible (APO-SUS) and apomorphine unsusceptible (APO-UNSUS) rats, originally selected on the basis of their behavioural response to the dopaminergic agonist apomorphine, were recently found to be genetically different in the number of gene copies of a component of the gamma-secretase complex called Aph-1b. Whereas APO-UNSUS rats have three copies of the gene, APO-SUS rats have either 1 or 2 copies. In addition we have shown that these rats show differences in cocaine and alcohol self administration, and that both early life events and late environmental factors can alter this self-administration behaviour. Thus, the data so far support the hypothesis that the APO-SUS and APO-UNSUS rats offer an interesting animal model for drug dependence in which genes and environment interact. We finally propose a theoretical model which can explain this gene-environment interaction. PMID- 16253226 TI - Vasodilatory and anti-inflammatory effects of the 1,2,3,4,6-penta-O-galloyl-beta D-glucose (PGG) via a nitric oxide-cGMP pathway. AB - Vasorelaxant and anti-inflammatory effects of a 1,2,3,4,6-penta-O-galloyl-beta-d glucose (PGG) isolated from the root barks of Paeonia suffruticosa and possible mechanisms responsible were investigated. PGG induced a concentration-dependent relaxation of the phenylephrine-precontracted rat aorta. This effect disappeared with the removal of functional endothelium. Pretreatment of the aortic tissues with either N(G)-nitro-L-arginine methyl ester (L-NAME) or 1H-[1,2,4]-oxadiazole [4,3-alpha]-quinoxalin-1-one (ODQ) inhibited the relaxation induced by PGG. Incubation of human umbilical vein endothelial cells (HUVECs) or carotid arteries isolated from rats with PGG increased the production of cGMP in a dose-dependent manner, but this effect was blocked by pretreatment with L-NAME and ODQ, respectively. PGG treatment attenuated tumor necrosis factor-alpha (TNF-alpha) induced nuclear factor-kappaB (NF-kappaB) p65 translocation in human umbilical vein endothelial cells. In addition, PGG suppressed the expression levels of adhesion molecules including intracellular cell adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) induced by TNF-alpha. TNF-alpha induced monocyte chemoattractant protein-1 (MCP-1) expression was also attenuated by addition of PGG. PGG treatment inhibited cellular adhesion of U937 cells onto human umbilical vein endothelial cells induced by TNF-alpha. Taken together, the present study suggests that PGG dilates vascular smooth muscle and suppresses the vascular inflammatory process via endothelium-dependent nitric oxide (NO)/cGMP signaling. PMID- 16253228 TI - Neural substrates of cocaine-cue associations that trigger relapse. AB - Learned associations that occur during the process of repeated drug use in addiction can later manifest as trigger factors in relapse to renewed drug seeking and drug-taking behavior. The process of conditioned-cued relapse of drug seeking behavior has been successfully modeled in animals using the reinstatement procedure, in which chronic drug self-administration can be extinguished or withheld, and then reinstated using conditioned stimuli previously paired with the drug. Our laboratory has extensively studied the neural circuitry underlying conditioned-cued drug-seeking during the expression of reinstatement. In order to study the learning process of drug-cue pairings, we further developed a procedure whereby discrete cocaine-cue pairings can be conducted in a single pavlovian training session in animals previously trained to self-administer cocaine. Presentation of these cues during later reinstatement trials produces robust responding over extinction levels at levels similar to those seen when animals experience the cues on a daily basis. In a series of experiments, we have shown that reversible pharmacological inactivation of the basolateral complex of the amygdala just prior to acquisition of cocaine-cue associations blocks the ability of cocaine-paired stimuli to elicit conditioned-cued reinstatement. This learning process is mediated in part by muscarinic acetylcholine and dopaminergic inputs to the basolateral complex of the amygdala, as intra-amygdala infusion of selective receptor antagonists at the time of acquisition significantly affects reinstatement. We have also recently found that disruption of neural activity within the basolateral complex of the amygdala at the time of consolidation (just after cocaine-cue pairings) will disrupt reinstatement. Taken together, these results reveal the importance of the amygdala in the acquisition, consolidation, and expression of drug-stimulus learning that drives relapse to drug-seeking behavior. PMID- 16253230 TI - Effect of spermine and N1-dansyl-spermine on epileptiform activity in mouse cortical slices. AB - N(1)-dansyl-spermine is a novel polyamine analogue, which has been demonstrated to have an antagonist action at the stimulatory polyamine site on the N-methyl-D aspartate (NMDA) receptor macrocomplex. Cortical wedges from genetically epilepsy prone DBA/2 mice demonstrate spontaneous epileptiform activity when perfused with Mg(2+)-free artificial cerebrospinal fluid (aCSF). This epileptiform activity has been demonstrated to be primarily mediated through the NMDA receptor. N(1)-dansyl spermine reduced the spontaneous epileptiform activity at a high dose (100 microM) but had no effect at a lower dose (50 microM). The polyamine, spermine (300 microM) caused an increase in the rate of the spontaneous epileptiform discharges. This effect of spermine was antagonised by administration of the low dose of N(1)-dansyl-spermine (50 microM). This further demonstrates the role of the NMDA receptor in the production of spontaneous epileptiform discharges in the cortical wedge preparation and clearly illustrates both the facilitatory action of spermine and the polyamine antagonist action of N(1)-dansyl-spermine at the stimulatory polyamine site on the NMDA receptor. PMID- 16253231 TI - Antidepressant treatments and human aggression. AB - Aggressive behaviour is associated with negative mood and poor impulse control. Serotonin has been specifically associated with impulse regulation and deficiencies in serotonin have been linked to impulsive aggression. However, aggression occurs in a social context and noradrenaline has been implicated in social motivation. Both serotonergic and noradrenergic antidepressants may therefore be effective in reducing aggression. The evidence for the effects of antidepressants on aggression comes from a wide range of sources but there are few controlled trials or experimental studies. Current findings point to decreases in negative mood and anger attacks and positive changes in personality traits after antidepressant treatment. Clinical studies in personality disorder patients have shown some efficacy for serotonergic antidepressants in reducing irritability and impulsive aggression. Experimental work in healthy volunteers has shown both serotonergic and noradrenergic antidepressants to increase assertiveness and affiliative behaviour. Both may therefore decrease aggression through different routes. PMID- 16253229 TI - (2E,6Z,10E)-7-hydroxymethyl-3,11,15-trimethyl-2,6,10,14-hexadecatetraen-1-ol (Plaunotol) increases cyclooxygenase-2 expression via nuclear factor kappaB and cyclic AMP response element in rat gastric epithelial cells. AB - Plaunotol, [(2E,6Z,10E)-7-hydroxymethyl-3,11,15-trimethyl-2,6,10,14 hexadecatetraen-1-ol], a gastroprotective agent, increases the prostaglandin production in the gastric mucosa and accelerates ulcer healing. The precise mechanisms underlying the gastroprotective actions by plaunotol are not known. On the other hand, cyclooxygenase (COX)-2 is a key enzyme in PGE(2) production and its induction is thought to have an important role in ulcer healing. We investigated the mechanism of plaunotol-mediated COX-2 induction in rat gastric epithelial (RGM1) cells. We used a PGE(2) enzyme-linked immunoassay kit and Western blot analysis to measure PGE(2) production and COX-2 induction with plaunotol treatment in serum-starved RGM1 cells. In addition, gel-shift assay, Western blot analysis and a reporter assay were performed to observe which Cox-2 promoter was involved in plaunotol-induced Cox-2 expression. The findings indicated that plaunotol treatment dose-dependently increased COX-2 expression and PGE(2) production. The nuclear factor kappaB (NF-kappaB) and cyclic AMP response element (CRE) sites of the COX-2 gene promoter were critical to plaunotol-mediated COX-2 expression. In conclusion, plaunotol induced COX-2 expression and increased PGE(2) production in serum-starved RGM1 cells via activation of the NF-kappaB and CRE sites of Cox-2 gene promoters. PMID- 16253232 TI - Selective in vivo anti-inflammatory action of the galactolipid monogalactosyldiacylglycerol. AB - The thermophilic blue-green alga ETS-05 colonises the therapeutic thermal muds of Abano and Montegrotto, Italy. Following the isolation, purification and identification of monogalactosyldiacylglycerol (MGDG), digalactosyldiacylglycerol (DGDG), sulphoquinovosyldiacylglycerol (SQDG) and phosphatidylglycerol from ETS 05, we here examine their in vivo anti-inflammatory activities. MGDG, DGDG and SQDG inhibit croton-oil-induced ear oedema in the mouse in a dose-dependent manner. Inhibition by MGDG is greater than that of the reference drug, betamethasone 17,21-dipropionate, and is largely abrogated following acyl group saturation. SQDG is the least potent of these glycoglycerolipids, and shows an early transient effect. In the in vivo carrageenan-induced paw oedema model in the mouse, the inhibitory effects are again dose dependent, with an enhanced efficacy of MGDG over DGDG, SQDG and the reference drug, indomethacin. These compounds are all less toxic than indomethacin. The selective and enhanced inhibitory effects of MGDG over DGDG indicate the mechanisms behind these in vivo anti-inflammatory actions. PMID- 16253234 TI - Novel pharmacotherapeutic targets for the management of drug addiction. AB - Despite individual variation in the liability to the abuse of psychoactive substances, there is substantial commonality shared by drugs of abuse. The knowledge of these common mechanisms together with the continued elucidation of the neurobiological underpinnings of withdrawal symptoms, drug intake, craving, relapse, and co-morbid psychiatric associations are critically important for the development of new therapeutic strategies. The present review will focus on recent advances in the development of innovative pharmacotherapeutic agents, which should promote higher efficacy (abstinence, prevention of relapse, long term recovery) and patient compliance, as well as improved safety profiles. PMID- 16253233 TI - Effect of chymase on intraocular pressure in rabbits. AB - Chymase is a chymotrypsin-like serine protease that is stored exclusively in the secretory granules of mast cells and converts big endothelins to endothelin-1 (1 31). The aim of this study was to evaluate the effect of chymase on intraocular pressure in rabbits. Chymase injection (3 and 10 mU) resulted in a trend toward increased intraocular pressure and a significant increase in intraocular pressure at a dose of 10 mU compared with the control. A specific chymase inhibitor, Suc Val-Pro-Phe(P)(OPh)(2), attenuated the ocular hypertension induced by chymase. Endothelin-1 (1-31) also caused ocular hypertension, which was inhibited by a selective endothelin ET(A) receptor antagonist, cyclo(D-Asp-Pro-D-Val-Leu-D-Trp) (BQ-123). Moreover, chymase-induced ocular hypertension was inhibited by BQ-123. These results suggest that chymase influences the regulation of intraocular pressure, and it is likely that the formation of endothelin-1 (1-31) and subsequent activation of endothelin ET(A) receptors are involved in the development of ocular hypertension induced by chymase. PMID- 16253235 TI - Endotoxin can decrease isolated rat parotid acinar cell amylase secretion in a nitric oxide-independent manner. AB - Salivary mucus and amylase have an anti-bacterial nature. Bacterial endotoxin is considered to decrease mucus secreting cell activity by nitric oxide-dependent mechanisms. In this study, the actions of endotoxin on amylase secreting cell activity have been studied. Endotoxin (Escherichia coli lipopolysaccharide; 3 mg/kg, i.v., 5 h) evoked nitric oxide synthase 2 (NOS2) induction in the rat whole parotid tissue (assessed by Western blot and the citrulline assay) and in rat isolated parotid acinar cells (assessed by Western blot and immunohistochemistry), and reduced basal and acetylcholine-stimulated amylase secretion from these isolated cells. However, N(G)-nitro-L-arginine methyl ester (0.1 mg/ml, 4 days in drinking water, yielding a dose of 25 mg/kg/day) did not affect amylase release under basal or acetylcholine-stimulated conditions, either in control acinar cells or those from endotoxin challenged rats. Thus, basal, acetylcholine-evoked or endotoxin-decreased cellular amylase secretion from rat isolated parotid acinar cells does not appear to be modulated by endogenous nitric oxide. PMID- 16253236 TI - Repeated administration of citalopram and imipramine alters the responsiveness of rat hippocampal circuitry to the activation of 5-HT7 receptors. AB - The effects of a selective serotonin reuptake inhibitor, citalopram, and a tricyclic antidepressant drug, imipramine, administered repetitively for 14 days, were investigated ex vivo in rat hippocampal slices. Spontaneous epileptiform bursts were recorded from the CA3 area in nominally Mg(2+)-free incubation conditions. 5-carboxamidotryptamine (5-CT) dose-dependently increased bursting frequency in the presence of N-[2-[4-(2-methoxyphenyl)-1 piperazinyl]ethyl]-N-2 pyridinylcyclohexanecarboxamide (WAY 100635). This effect could be dose dependently blocked by (2R)-1-[(3-Hydroxyphenyl)sulfonyl]-2-[2-(4-methyl-1 piperidinyl)ethyl]pyrrolidine hydrochloride (SB 269970), thus implicating the involvement of 5-HT(7) receptors. Repeated treatment with citalopram or imipramine resulted in an attenuation of the excitatory effects of the activation of hippocampal 5-HT(7) receptor. PMID- 16253237 TI - Hemodynamic effects of combined sildenafil and L-arginine during acute pulmonary embolism-induced pulmonary hypertension. AB - Sildenafil attenuates acute pulmonary embolism-induced pulmonary hypertension. However, the hemodynamic effects of sildenafil in combination with other vasodilators during acute pulmonary embolism have not been examined yet. In the present study, we examined the hemodynamic effects of combined sildenafil (0.25 mg/kg, i.v.) and L-arginine (100, 200, 500, and 1000 mg/kg/h, i.v.) in an anesthetized dog model of acute pulmonary embolism. Plasma nitrite/nitrate (NO(x)) and cGMP concentrations were determined using an ozone-based chemiluminescence assay and a commercial enzyme immunoassay, respectively. We found that L-arginine alone did not attenuate acute pulmonary embolism-induced pulmonary hypertension. However, significant decreases in mean pulmonary artery pressure were observed 30, 45, 60, and 75 min after the administration of sildenafil alone or after the combined administration of sildenafil and L arginine (all P < 0.05). No significant differences among groups were observed in the respiratory parameters. While L-arginine significantly increased NO(x) concentrations, cGMP concentrations increased only when sildenafil was administered (all P < 0.05). These results suggest that while sildenafil attenuates acute pulmonary embolism-induced pulmonary hypertension, L-arginine does not enhance the beneficial hemodynamic effects of sildenafil. In addition, these findings suggest that stimulation of NO synthesis with L-arginine during acute pulmonary embolism does not produce beneficial effects. PMID- 16253238 TI - Eckol isolated from Ecklonia cava attenuates oxidative stress induced cell damage in lung fibroblast cells. AB - We have investigated the cytoprotective effect of eckol, which was isolated from Ecklonia cava, against oxidative stress induced cell damage in Chinese hamster lung fibroblast (V79-4) cells. Eckol was found to scavenge 1,1-diphenyl-2 picrylhydrazyl radical, hydrogen peroxide (H(2)O(2)), hydroxy radical, intracellular reactive oxygen species (ROS), and thus prevented lipid peroxidation. As a result, eckol reduced H(2)O(2) induced cell death in V79-4 cells. In addition, eckol inhibited cell damage induced by serum starvation and radiation by scavenging ROS. Eckol was found to increase the activity of catalase and its protein expression. Further, molecular mechanistic study revealed that eckol increased phosphorylation of extracellular signal-regulated kinase and activity of nuclear factor kappa B. Taken together, the results suggest that eckol protects V79-4 cells against oxidative damage by enhancing the cellular antioxidant activity and modulating cellular signal pathway. PMID- 16253239 TI - ET-18-O-CH3-induced apoptosis is causally linked to COX-2 upregulation in H-ras transformed human breast epithelial cells. AB - Abnormally elevated expression of cyclooxygenase-2 (COX-2) has been frequently observed in transformed or malignant cells, and certain non-steroidal anti inflammatory drugs with COX-2 inhibitory activity exert anti-neoplastic or chemopreventive effects. Contrary to this notion, we have found that a novel alkylphospholipid type antitumor agent ET-18-O-CH3 (1-O-octadecyl-2-O-methyl glycero-3-phosphocholine) induces COX-2 expression in H-ras transformed human breast epithelial cells (MCF10A-ras) while it causes apoptosis at the same concentration range. The addition of a selective COX-2 inhibitor SC-58635 and COX 2 gene knock down with the siRNA blocked ET-18-O-CH3-induced apoptosis, suggesting that COX-2 induction by this drug is causally linked to its apoptosis inducing activity. ET-18-O-CH3 enhanced the transcriptional activities of cyclic AMP response element which is a key regulator of COX-2 expression. 15-Deoxy Delta(12,14) prostaglandin J2 is, an endogenous ligand for peroxisome proliferator-activated receptor gamma (PPARgamma), has been known to possess proapoptotic potential in diverse cell types. ET-18-O-CH3 treatment resulted in elevated release of 15d-PGJ2 and DNA binding and transcriptional activity of PPARgamma. Based on these findings, it is likely that ET-18-O-CH3 induces COX-2 expression and production of 15d-PGJ2 which may mediate the ET-18-O-CH3-induced apoptosis in MCF10A-ras cells. PMID- 16253240 TI - Desumoylation of homeodomain-interacting protein kinase 2 (HIPK2) through the cytoplasmic-nuclear shuttling of the SUMO-specific protease SENP1. AB - The modification of homeodomain-interacting protein kinase 2 (HIPK2) by small ubiquitin-like modifier 1 (SUMO-1) plays an important role in its targeting into the promyelocytic leukemia body, as well as in its differential interaction with binding partner, but the desumoylation of HIPK2 by SUMO-specific proteases is largely unknown. In this study, we show that HIPK2 is a desumoylation target for the SUMO-specific protease SENP1 that shuttles between the cytoplasm and the nucleus. Mutation analyses reveal that SENP1 contains the nuclear export sequence (NES) within the extreme carboxyl-terminal region, and SENP1 is exported to the cytoplasm in a NES-dependent manner. Sumoylated HIPK2 are deconjugated by SENP1 both in vitro and in cultured cells, and the desumoylation is enhanced either by the forced translocation of SENP1 into the nucleus or by the SENP1 NES mutant. Concomitantly, desumoylation induces dissociation of HIPK2 from nuclear bodies. These results demonstrate that HIPK2 is a target for SENP1 desumoylation, and suggest that the desumoylation of HIPK2 may be regulated by the cytoplasmic nuclear shuttling of SENP1. PMID- 16253241 TI - Transgenic tobacco plants overexpressing glyoxalase enzymes resist an increase in methylglyoxal and maintain higher reduced glutathione levels under salinity stress. AB - The mechanism behind enhanced salt tolerance conferred by the overexpression of glyoxalase pathway enzymes was studied in transgenic vis-a-vis wild-type (WT) plants. We have recently documented that salinity stress induces higher level accumulation of methylglyoxal (MG), a potent cytotoxin and primary substrate for glyoxalase pathway, in various plant species [Yadav, S.K., Singla-Pareek, S.L., Ray, M., Reddy, M.K. and Sopory, S.K. (2005) MG levels in plants under salinity stress are dependent on glyoxalase I and glutathione. Biochem. Biophys. Res. Commun. 337, 61-67]. The transgenic tobacco plants overexpressing glyoxalase pathway enzymes, resist an increase in the level of MG that increased to over 70% in WT plants under salinity stress. These plants showed enhanced basal activity of various glutathione related antioxidative enzymes that increased further upon salinity stress. These plants suffered minimal salinity stress induced oxidative damage measured in terms of the lipid peroxidation. The reduced glutathione (GSH) content was high in these transgenic plants and also maintained a higher reduced to oxidized glutathione (GSH:GSSG) ratio under salinity. Manipulation of glutathione ratio by exogenous application of GSSG retarded the growth of non transgenic plants whereas transgenic plants sustained their growth. These results suggest that resisting an increase in MG together with maintaining higher reduced glutathione levels can be efficiently achieved by the overexpression of glyoxalase pathway enzymes towards developing salinity stress tolerant plants. PMID- 16253242 TI - B7-H1 (CD274) inhibits the development of herpetic stromal keratitis (HSK). AB - The co-signaling molecule B7-H1 (CD274) functions as both a co-inhibitor through programmed death-1 (PD-1) receptor and a co-stimulator via an as-yet-unidentified receptor on T cells. We investigated the physiological role of endogenous B7-H1 in the pathogenesis of herpetic stromal keratitis (HSK) caused by herpes simplex virus type 1 (HSV-1). Following HSV-1 infection of the cornea of mice, B7-H1 expression was up-regulated in the CD11b+ macrophage population in the draining lymph nodes (dLN) and in the inflamed cornea. In addition, HSV-1 infection significantly increased PD-1 expression on CD4+ T cells in the dLN and inflamed cornea. The administration of antagonistic B7-H1 monoclonal antibody resulted in the proliferation of HSV-specific CD4+ T cells that secreted interferon (INF) gamma, and inhibited the apoptosis of HSV-specific CD4+ T cells, which exaggerated HSK. These results strongly suggest that the B7-H1 may be involved in suppression of the development of HSK. PMID- 16253243 TI - Focal adhesion molecules as potential target of lead toxicity in NRK-52E cell line. AB - In this study, we investigated the influence of inorganic lead (Pb(II)), an environmental pollutant having nephrotoxic action, on the focal adhesion (FA) organization of a rat kidney epithelial cell line (NRK-52E). In particular, we evaluated the effects of the metal on the recruitment of paxillin, focal adhesion kinase, vinculin and cytoskeleton proteins at the FAs complexes. We provided evidences that, in proliferating NRK-52E cell cultures, low concentrations of Pb(II) affect the cell adhesive ability and stimulate the disassembly of FAs, thus inhibiting the integrin-activated signalling. These effects appeared to be strictly associated to the Pb-induced arrest of cell cycle at G0/G1 phase also proved in this cell line. PMID- 16253244 TI - The haloperoxidase of the agaric fungus Agrocybe aegerita hydroxylates toluene and naphthalene. AB - The mushroom Agrocybe aegerita secretes a peroxidase (AaP) that catalyzes halogenations and hydroxylations. Phenol was brominated to 2- and 4-bromophenol (ratio 1:4) and chlorinated to a lesser extent to 2-chlorophenol. The purified enzyme was found to oxidize toluene via benzyl alcohol and benzaldehyde into benzoic acid. A second fraction of toluene was hydroxylated to give p-cresol as well as o-cresol and methyl-p-benzoquinone. The UV-Vis absorption spectrum of purified AaP showed high similarity to a resting state cytochrome P450 with the Soret band at 420 nm and additional maxima at 278, 358, 541 and 571 nm; the AaP CO-complex had a distinct absorption maximum at 445 nm that is characteristic for heme-thiolate proteins. AaP regioselectively hydroxylated naphthalene to 1 naphthol and traces of 2-naphthol (ratio 36:1). H2O2 was necessarily required for AaP function and hence the hydroxylations catalyzed by AaP can be designated as peroxygenation and the enzyme as an extracellular peroxygenase. PMID- 16253245 TI - Transgenic mice expressing bovine PrP with a four extra repeat octapeptide insert mutation show a spontaneous, non-transmissible, neurodegenerative disease and an expedited course of BSE infection. AB - Transgenic (Tg) mice carrying four extra octapeptide repeats (OR) in the bovine PrP gene (10OR instead of 6) have been generated. In these mice, neuropathological changes were observed depending upon the level of transgene expression. These changes primarily involved a slowly advancing neurological disorder, characterized clinically by ataxia, and neuropathologically, by vacuolization in different brain areas, gliosis, and loss of cerebellar granule cells. Accumulation of insoluble bovine 10OR-PrP (bo10OR-PrP) was observed depending on the level of expression but no infectivity was found associated with this insoluble form. We also compared the behavior of bo6OR-PrP and bo10OR-PrP Tg mouse lines in response to BSE infection. BSE-inoculated bo10ORTg mice showed an altered course of BSE infection, reflected by reduced incubation times when compared to bo6ORTg mice expressing similar levels of the wild type 6OR-PrP. In BSE-inoculated mice, it was possible to detect PrP(res) in 100% of the animals. While insoluble bo10OR-PrP from non-inoculated bo10ORTg mice was non-infectious, brain homogenates from BSE-inoculated bo10ORTg mice were highly infectious in all the Tg mouse lines tested. This Tg mouse model constitutes a new way of understanding the pathobiology of bovine transmissible spongiform encephalopathy. Its potential applications include the assessment of new therapies against prion diseases. PMID- 16253246 TI - Nitric oxide induces tau hyperphosphorylation via glycogen synthase kinase-3beta activation. AB - Nitric oxide is associated with neurofibrillary tangle, which is composed mainly of hyperphosphorylated tau in the brain of Alzheimer's disease (AD). However, the role of nitric oxide in tau hyperphosphorylation is unclear. Here we show that nitric oxide produced by sodium nitroprusside (SNP), a recognized donor of nitric oxide, induces tau hyperphosphorylation at Ser396/404 and Ser262 in HEK293/tau441 cells with a simultaneous activation of glycogen synthase kinase-3beta (GSK 3beta). Pretreatment of the cells with 10 mM lithium chloride (LiCl), an inhibitor of GSK-3, 1 h before SNP administration inhibits GSK-3beta activation and prevents tau from hyperphosphorylation. This is the first direct evidence demonstrating that nitric oxide induces AD-like tau hyperphosphorylation in vitro, and GSK-3beta activation is partially responsible for the nitric oxide induced tau hyperphosphorylation. It is suggested that nitric oxide may be an upstream element of tau abnormal hyperphosphorylation in AD. PMID- 16253247 TI - The hairy and enhancer of split 1 is a negative regulator of the repressor element silencer transcription factor. AB - Silencing of the transcriptional repressor REST is required for terminal differentiation of neuronal and beta-cells. In this study, we hypothesized that REST expression is controlled by hairy and enhancer of split 1 (HES-1), a transcriptional repressor that plays an important role in brain and pancreas development. We identified several N elements (CTNGTG) within the promoter of REST and confirmed that HES-1 associates with the endogenous promoter of REST. Moreover, using a cells model that overexpress HES-1 and a combination of experimental approaches, we demonstrated that HES-1 reduces endogenous REST expression. Taken together, these results indicate that HES-1 is an upstream negative regulator of REST expression. PMID- 16253248 TI - Claudins upregulation in human colorectal cancer. AB - In colorectal cancer tight junction molecular and morphological alterations are poorly understood. In this study, adenocarcinoma tissues and their paired normal mucosa (n = 12) were analyzed for tight junction alterations molecular. The expression of claudin-1, -3 and -4 was upregulated 5.7-, 1.5- and 2.4-fold, respectively, in colorectal tumor tissues in comparison to the normal ones. Although tight junction remains in the cancerous epithelium, its barrier function was altered. Despite claudins overexpression, paracellular permeability to ruthenium red was increased and a significant disorganization of tight junction strands was observed in freeze fracture replicas. Whereas the functional significance of claudin overexpression in colorectal cancer is unclear, these proteins can become potential markers and targets in colorectal cancer. PMID- 16253249 TI - The acute stress response of red porgy, Pagrus pagrus, kept on a red or white background. AB - The skin colour of red porgy, Pagrus pagrus, can be modified by exposure to different background colours. Red and white background colours brighten the dark skin colour that develops under common culture conditions in red porgy. To assess whether skin colour is also modified by aquaculture related handling stress, we subjected red porgy to 5 min of netting stress combined with air exposure. Fish kept on a white background have: (1) a lighter skin colour, which is not influenced by an acute stressor, (2) a less saturated red colour, which significantly decreases 24h post-handling, and (3) a similar hue as fish kept on a red background. The first plasma parameters to rise after application of the stressor are cortisol, lactate and Na(+); then, glucose levels rose. Other plasma ions (Ca(2+), Cl(-), K(+)) were not affected up to 2h post-stressor, but had decreased at 8 and 24h after handling. Plasma pH decreased over the first 2h post handling, indicative of plasma acidosis upon air exposure. The acidosis then coincided with increases in plasma lactate levels. As alphaMSH levels were not significantly affected by the stressor while cortisol levels showed a five to tenfold increase, we suggest that following acute stress in red porgy, plasma cortisol release is controlled by ACTH, perhaps in combination with a sympathic stimulation. PMID- 16253250 TI - Characterization of turkey and chicken ghrelin genes, and regulation of ghrelin and ghrelin receptor mRNA levels in broiler chickens. AB - Ghrelin, a peptide hormone produced by the stomach in mammals, stimulates growth hormone release and food intake. Recently, ghrelin was identified and characterized in chicken proventriculus and shown to stimulate growth hormone release but inhibit feed intake. The purpose of this work was to identify and further characterize the ghrelin gene in chickens and in turkeys. Using molecular cloning techniques we have sequenced cDNAs corresponding to chicken (White Leghorn) and turkey ghrelin mRNAs. A total of 844 (chicken) or 869 (turkey) bases including the complete coding regions (CDS), and the 5'- and 3'-untranslated regions (UTRs) were determined. Nucleotide sequence (CDS) predicted a 116 amino acid precursor protein (preproghrelin) for both the chicken and the turkey that demonstrated complete conservation of an N-terminal 'active core' (GSSF) including a serine (position 3 of the mature hormone) known to be a modification (acylation) site important for ghrelin bioactivity. Additional nucleotide sequence was found in the 5'-UTRs of both Leghorn and turkey cDNAs that was not present in broilers or the red jungle fowl. The turkey ghrelin gene, sequenced from genomic DNA templates, contained five exons and four introns, a structure similar to mammalian and chicken ghrelin genes. Ghrelin was highly expressed in proventriculus with much lower levels of expression in other tissues such as pancreas, brain, and intestine. RT-PCR was used to quantify ghrelin mRNA levels relative to 18S rRNA in 3-week-old male broiler chickens. The level of ghrelin mRNA increased in proventriculus in response to fasting but did not decline with subsequent refeeding. Plasma ghrelin levels did not change significantly in response to fasting or refeeding and did not appear to reflect changes in proventriculus ghrelin mRNA levels. Ghrelin mRNA levels declined in broiler pancreas after a 48 h fast and increased upon refeeding. Expression of the gene encoding the receptor for ghrelin (growth hormone secretagogue receptor, GHS-R) and a variant form was detected in a variety of tissues collected from 3-week-old male broiler chickens possibly suggesting autocrine/paracrine effects. These results offer new information about the avian ghrelin and ghrelin receptor genes and the potential role that this system might play in regulating feed intake and energy balance in poultry. PMID- 16253251 TI - Salvage of osteoporotic ankle fractures after failed primary fixation with an ankle arthrodesis nail: A report on four cases. PMID- 16253252 TI - Failure of reamed nailing in humeral non-union: an analysis of 26 patients. PMID- 16253253 TI - Dynamic hip screw in the management of reverse obliquity intertrochanteric neck of femur fractures [Injury 2005;36:105-9]. PMID- 16253254 TI - Tension-free vaginal tape for elderly women with stress urinary incontinence. AB - OBJECTIVE: To investigate the efficacy of the tension-free vaginal tape (TVT) procedure for the management of stress urinary incontinence (SUI) in elderly women. METHOD: A total of 55 women aged between 65 and 86 years underwent a TVT procedure for urodynamic SUI. Of these, 15 (27%) had undergone previous surgery for treatment of SUI. Before the TVT procedure, a complete medical history was taken and a gynecologic examination performed. RESULTS: Operating time ranged between 11 and 35 min (excluding the time of concomitant surgery, if any); hospitalization time ranged between 1 and 5 days; and no severe intraoperative or postoperative complications occurred. Cure occurred in 39 (76%) of 51 evaluable patients and its rate was positively associated with bladder neck mobility. Among patients in whom the angle of displacement on the Q-tip test was less than 30 degrees , 42% became continent whereas among those in whom it was 30 degrees or higher, 90% became continent (P<.001). Among those in whom the angle was between 20 degrees and 30 degrees, 57% became continent, and among those in whom it was less than 10 degrees, 80% remained incontinent. CONCLUSION: The TVT procedure in elderly women with SUI offers a satisfactory cure rate; however, in patients with significantly decreased bladder neck mobility (an angle <20 degrees on the Q-tip test), the results are not encouraging. PMID- 16253255 TI - Effect of acupuncture on labor. PMID- 16253256 TI - Extra-amniotic saline infusion vs. cervical traction with Foley catheter for second-trimester pregnancy termination. PMID- 16253257 TI - Rectovaginal fistula repair using fascia graft of autologous abdominal muscles. PMID- 16253258 TI - Persistent scalp bleeding due to fetal coagulopathy following fetal blood sampling. PMID- 16253259 TI - Thriving in nursing homes in Norway: contributing aspects described by residents. AB - BACKGROUND AND OBJECTIVES: Knowledge about residents' perception of what contributes to well-being and thriving in nursing homes is scarce. The aim of this study was to investigate mentally lucid residents' perspective on what contributes to thriving in a nursing home. DESIGN: A qualitative study with a descriptive-exploratory design. SETTINGS: Two nursing homes in Norway. PARTICIPANTS: 26 mentally lucid nursing home residents. METHODS: Data collection comprised participant observation and open-ended interviews. RESULTS: Two core aspects contributing to thriving were identified: The residents' attitude towards living in a nursing home and the quality of care and caregivers. The residents' attitude was the innermost core aspect. Five additional aspects contributing to thriving were identified: Positive peer relationships, participation in meaningful activities, opportunities to go outside the ward or nursing home, positive relationships with family, and qualities in the physical environment. CONCLUSION: Several factors contribute to an experience of thriving. The findings challenge the 'traditional' passive role of residents by documenting their active contributions to their level of thriving in a nursing home. PMID- 16253260 TI - Development and evaluation of a multifaceted ergonomics program to prevent injuries associated with patient handling tasks. AB - PROBLEM STATEMENT: Nurses have one of the highest rates of work-related musculoskeletal injury of any profession. Over the past 30 years, efforts to reduce work-related musculoskeletal disorders in nurses have been largely unsuccessful. SPECIFIC AIMS: The primary goal of this program was to create safer working environments for nursing staff who provide direct patient care. Our first objective was to design and implement a multifaceted program that successfully integrated evidence-based practice, technology, and safety improvement. The second objective was to evaluate the impact of the program on injury rate, lost and modified work days, job satisfaction, self-reported unsafe patient handling acts, level of support for program, staff and patient acceptance, program effectiveness, costs, and return on investment. INTERVENTION: The intervention included six program elements: (1) Ergonomic Assessment Protocol, (2) Patient Handling Assessment Criteria and Decision Algorithms, (3) Peer Leader role, "Back Injury Resource Nurses", (4) State-of-the-art Equipment, (5) After Action Reviews, and (6) No Lift Policy. METHODS: A pre-/post design without a control group was used to evaluate the effectiveness of a patient care ergonomics program on 23 high risk units (19 nursing home care units and 4 spinal cord injury units) in 7 facilities. Injury rates, lost work days, modified work days, job satisfaction, staff , and patient acceptance, program effectiveness, and program costs/savings were compared over two nine month periods: pre-intervention (May 2001-January 2002) and post-intervention (March 2002-November 2002). Data were collected prospectively through surveys, weekly process logs, injury logs, and cost logs. RESULTS: The program elements resulted in a statistically significant decrease in the rate of musculoskeletal injuries as well as the number of modified duty days taken per injury. While the total number of lost workdays decreased by 18% post-intervention, this difference was not statistically significant. There were statistically significant increases in two subscales of job satisfaction: professional status and tasks requirements. Self-reports by nursing staff revealed a statistically significant decrease in the number of 'unsafe' patient handling practices performed daily. Nurses ranked program elements they deemed to be "extremely effective": equipment was rated as most effective (96%), followed by No Lift Policy (68%), peer leader education program (66%), ergonomic assessment protocol (59%), patient handling assessment criteria and decision algorithms (55%), and lastly after action reviews (41%). Perceived support and interest for the program started at a high level for managers and nursing staff and remained very high throughout the program implementation. Patient acceptance was moderate when the program started but increased to very high by the end of the program. Although the ease and success of program implementation initially varied between and within the facilities, after six months there was strong evidence of support at all levels. The initial capital investment for patient handling equipment was recovered in approximately 3.75 years based on annual post-intervention savings of over $200,000/year in workers' compensation expenses and cost savings associated with reduced lost and modified work days and worker compensation. CONCLUSIONS: This multi-faceted program resulted in an overall lower injury rate, fewer modified duty days taken per injury, and significant cost savings. The program was well accepted by patients, nursing staff, and administrators. Given the significant increases in two job satisfaction subscales (professional status and task requirements), it is possible that nurse recruitment and retention could be positively impacted. PMID- 16253261 TI - Attitudes toward Web-based distance learning among public health nurses in Taiwan: a questionnaire survey. AB - BACKGROUND: Public health nurses (PHNs) often cannot receive in-service education due to limitations of time and space. Learning through the Internet has been a widely used technique in many professional and clinical nursing fields. The learner's attitude is the most important indicator that promotes learning. OBJECTIVES: The purpose of this study was to investigate PHNs' attitude toward web-based learning and its determinants. DESIGN: This study conducted a cross sectional research design. SETTINGS: 369 health centers in Taiwan. PARTICIPANTS: The population involved this study was 2398 PHNs, and we used random sampling from this population. Finally, 329 PHNs completed the questionnaire, with a response rate of 84.0%. METHODS: Data were collected by mailing the questionnaire. RESULTS: Most PHNs revealed a positive attitude toward web-based learning (mean+/-SD=55.02+/-6.39). PHNs who worked at village health centers, a service population less than 10,000, PHNs who had access to computer facility and on-line hardware in health centers and with better computer competence revealed more positive attitudes (p<0.01). CONCLUSIONS: Web-based learning is an important new way of in-service education; however, its success and hindering factors require further investigation. Individual computer competence is the main target for improvement, and educators should also consider how to establish a user friendly learning environment on the Internet. PMID- 16253262 TI - Characterization of smooth muscle-like cells in circulating human peripheral blood. AB - Smooth muscle cells play an important role in human vascular diseases. Several lines of evidence demonstrate that circulating smooth muscle precursor cells contribute to intimal hyperplasia in animal models. We obtained large spindle cells expressing alpha-smooth muscle actin (alpha-SMA), denoted here as "smooth muscle-like cells" (SMLC), from human peripheral blood mononuclear cells (PBMC). SMLC derived from human PBMC proliferated readily and expressed pro-inflammatory genes during early culture. After long-term culture, SMLC could contract and express characteristic smooth muscle cell markers. We found peripheral blood mononuclear cell expressing alpha-smooth muscle actin in the circulating blood that bore CD14 and CD105. Sorted CD14/CD105 double-positive PBMC could differentiate into SMLC. The number of CD14-CD105-bearing PBMC increased significantly in patients with coronary artery disease compared to patients without coronary artery disease. These results support the novel concept that smooth muscle precursor cells exist in circulating human blood and may contribute to the pathogenesis of vascular diseases. PMID- 16253263 TI - Reliability of segmental accelerations measured using a new wireless gait analysis system. AB - The purpose of this study was to determine the inter- and intra-examiner reliability, and stride-to-stride reliability, of an accelerometer-based gait analysis system which measured 3D accelerations of the upper and lower body during self-selected slow, preferred and fast walking speeds. Eight subjects attended two testing sessions in which accelerometers were attached to the head, neck, lower trunk, and right shank. In the initial testing session, two different examiners attached the accelerometers and performed the same testing procedures. A single examiner repeated the procedure in a subsequent testing session. All data were collected using a new wireless gait analysis system, which features near real-time data transmission via a Bluetooth network. Reliability for each testing condition (4 locations, 3 directions, 3 speeds) was quantified using a waveform similarity statistic known as the coefficient of multiple determination (CMD). CMD's ranged from 0.60 to 0.98 across all test conditions and were not significantly different for inter-examiner (0.86), intra-examiner (0.87), and stride-to-stride reliability (0.86). The highest repeatability for the effect of location, direction and walking speed were for the shank segment (0.94), the vertical direction (0.91) and the fast walking speed (0.91), respectively. Overall, these results indicate that a high degree of waveform repeatability was obtained using a new gait system under test-retest conditions involving single and dual examiners. Furthermore, differences in acceleration waveform repeatability associated with the reapplication of accelerometers were small in relation to normal motor variability. PMID- 16253264 TI - Microstructural insight into pedestrian pelvic fracture as assessed by high resolution computed tomography. AB - Pelvic and femoral neck bone surface strains were recorded in five full-body human cadaver vehicle-pedestrian impacts. Impacts were performed at 40 km/h using automotive front ends constructed to represent those used in previously reported finite element simulations. While experimental kinematics and bone strains closely matched model predictions, observed pelvic fractures did not consistently agree with the model, and could not be solely explained by vehicle geometry. In an attempt to reconcile injury outcome with factors apart from vehicle design, a proxy measure of subject skeletal health was assessed by high-resolution quantitative computed tomography (HRqCT) of the femoral neck. The incidence of hip/pelvis fracture was found to be consistent with low volumetric bone mineral density and low trabecular bone density. This finding lends quantitative support to the notion that healthy trabecular architecture is crucial in withstanding non physiological impact loads. Furthermore, it is recommended that injury criteria used to assess vehicle safety with regard to pedestrians consider the increased susceptibility of elderly victims to pelvic fracture. PMID- 16253265 TI - Nanoindentation hardness of mineralized tissues. AB - A series elastic and plastic deformation model [Sakai, M., 1999. The Meyer hardness: a measure for plasticity? Journal of Materials Research 14(9), 3630 3639] is used to deconvolute the resistance to plastic deformation from the plane strain modulus and contact hardness parameters obtained in a nanoindentation test. Different functional dependencies of contact hardness on the plane strain modulus are examined. Plastic deformation resistance values are computed from the modulus and contact hardness for engineering materials and mineralized tissues. Elastic modulus and plastic deformation resistance parameters are used to calculate elastic and plastic deformation components, and to examine the partitioning of indentation deformation between elastic and plastic. Both the numerical values of plastic deformation resistance and the direct computation of deformation partitioning reveal the intermediate mechanical responses of mineralized composites when compared with homogeneous engineering materials. PMID- 16253266 TI - Boric acid as a mobile phase additive for high performance liquid chromatography separation of ribose, arabinose and ribulose. AB - A new high performance liquid chromatographic (HPLC) method is described for the analysis of ribose, arabinose and ribulose mixtures obtained from (bio)chemical isomerization processes. These processes gain importance since the molecules can be used for the synthesis of antiviral therapeutics. The HPLC method uses boric acid as a mobile phase additive to enhance the separation on an Aminex HPX-87K column. By complexing with boric acid, the carbohydrates become negatively charged, thus elute faster from the column by means of ion exlusion and are separated because the complexation capacity with boric acid differs from one carbohydrate to another. Excellent separation between ribose, ribulose and arabinose was achieved with concentrations between 0.1 and 10 gL(-1) of discrete sugar. PMID- 16253267 TI - Comparison of two ELISAs for the determination of Hsp70 in serum. AB - We have compared a previously developed in-house Sandwich-ELISA with a commercial kit for the determination of heat shock protein (Hsp) 70 in serum. Samples from 64 participants were tested and there was a significant correlation between results obtained using the two assays (r = 0.807, p < 0.0001). Additionally, when ranking samples on a categorical scale, the agreement was good (72%). In the commercial test system Hsp70 was detectable in 42 (66%) of the sera, compared with 61 (95%) in the in-house ELISA method. The three samples with undetectable levels of Hsp70 in the in-house ELISA were among the 22 samples with undetectable levels of Hsp70 in the commercial ELISA kit. The apparent serum concentrations detected were different in the two systems. This dissimilarity can be ascribed to differences in the matrix used. We conclude that the in-house ELISA is more economical and performs well when measuring physiologically high, as well as low, concentrations of Hsp70. PMID- 16253268 TI - Comparative study on the relationship between photoperiodic host-seeking behavioral patterns and the eye parameters of mosquitoes. AB - Relationships between the ommatidial structure and photoperiodic behavior of several mosquito species were investigated. Host-seeking behavioral patterns of mosquitoes were classified into four main groups based on previously compiled reports on field or laboratory biting activity. These groups were pattern I and I' (nocturnal), pattern II (crepuscular and nocturnal), pattern III (crepuscular and diurnal), and pattern IV (diurnal). Eye parameters (product of facet diameter and interommatidial angle) of mosquitoes that belong to the pattern I and I' group were higher (2.7-4.2) than those of mosquitoes that belong to the pattern IV group (0.8-2.3). Eye parameters of the mosquitoes categorized in the pattern II and III groups were intermediate (2.3-2.6). These results suggest that the crepuscular behavior of mosquitoes undergoes a transition in the course of evolution from nocturnal behavior to diurnal behavior. Large variations in the eye parameters were observed even within the same genus depending on their photoperiodic behavior. Therefore, the ommatidial structure of mosquitoes appears to be determined, not taxonomically, but evolutionarily by the photoenvironment in which the mosquitoes are most active. PMID- 16253269 TI - Genotypic analysis of Nomuraea rileyi collected from various noctuid hosts. AB - The genetic diversity of 79 Nomuraea rileyi isolates from various lepidopteran hosts in Asia, North America, and South America was evaluated using amplified fragment length polymorphism (AFLP) analysis. Cluster analysis separated the N. rileyi isolates into two major groups and seven subgroups. The resulting dendrogram generally classified the N. rileyi isolates based on insect host and geographical region. The haplotypic diversity index of N. rileyi subpopulations from each location and host origin was ranging from 0.8788 to 1.000. However, analysis of molecular variance (AMOVA) demonstrated no significant differences (p =0.3421) among N. rileyi isolates from different continents. Whereas the genetic variation among the N. rileyi populations from the different host insects within each continent was significantly different (p <0.0001). PMID- 16253270 TI - Analysis of putative RNase P RNA from orthopoxviruses. AB - A putative RNase P RNA gene in camelpox virus, one of the orthopoxviruses, was cloned and transcribed in vitro. No RNase P activity could be detected in vitro from camelpox virus RNase P RNA alone, or by addition of the Escherichia coli RNase P protein subunit to reaction mixtures. Camelpox virus RNase P RNA reconstituted in vitro with camel or HeLa cell extracts, which were pre-treated with micrococcal nuclease to degrade the endogenous RNase P RNA, showed no RNase P activity. Vaccinia virus, another orthopoxvirus, showed no RNase P activity in vaccinia-infected HeLa cells, even though transcription of the vaccinia RNase P RNA could be identified in the cells by both Northern blot and RNase protection assay. Camelpox virus RNase P RNA inhibited an endogenous HeLa RNase P activity by 20% in our assays. The 5S RNA showed no significant inhibition in this assay. PMID- 16253271 TI - Design and characterization of viral polypeptide inhibitors targeting Newcastle disease virus fusion. AB - Paramyxovirus infections can be detected worldwide with some emerging zoonotic viruses and currently there are no specific therapeutic treatments or vaccines available for many of these diseases. Recent studies have demonstrated that peptides derived from the two heptad repeat regions (HR1 and HR2) of paramyxovirus fusion proteins could be used as inhibitors of virus fusion. The mechanism underlying this activity is in accordance with that of class I virus fusion proteins, of which human immunodeficiency virus (HIV) and influenza virus fusion proteins are members. For class I virus fusion proteins, the HR1 fragment binds to HR2 to form a six-helix bundle with three HR1 fragments forming the central coiled bundle surrounded by three coiled HR2 fragments in the post fusion conformational state (fusion core). It is hypothesized that the introduced exogenous HR1 or HR2 can compete against their endogenous counterparts, which results in fusion inhibition. Using Newcastle disease virus (NDV) as a model, we designed several protein inhibitors, denoted HR212 as well asHR121 and 5-Helix, which could bind the HR1 or HR2 region of fusion protein, respectively. All the proteins were expressed and purified using a GST-fusion expression system in Escherichia coli. The HR212 or GST-HR212 protein, which binds the HR1 peptide in vitro, displayed inhibitory activity against NDV-mediated cell fusion, while the HR121 and 5-Helix proteins, which bind the HR2 peptide in vitro, inhibited virus fusion from the avirulent NDV strain when added before the cleavage of the fusion protein. These results showed that the designed HR212, HR121 or 5-Helix protein could serve as specific antiviral agents. These data provide additional insight into the difference between the virulent and avirulent strains of NDV. PMID- 16253272 TI - Structural basis for cooperative transcription factor binding to the CBP coactivator. AB - Regulation of transcription requires interactions between transcriptional activators and transcriptional co-activator CREB binding protein (CBP). The KIX domain of CBP can bind simultaneously to two different proteins, providing an additional mechanism for transcriptional regulation. Here we describe the solution structure of the ternary complex formed by cooperative binding of activation domains from the c-Myb and mixed lineage leukemia (MLL) transcription factors to the KIX domain. The MLL and c-Myb domains form helices that bind to two distinct hydrophobic grooves on opposite faces of KIX. Compared to the binary KIX:c-Myb complex, significant changes are observed in the structure of KIX at the MLL binding interface in the ternary complex. Two regions of KIX that are disordered in the binary complex become structured in the ternary complex: a flexible loop forms intimate contacts with bound MLL, and the C-terminal helix is extended and stabilized by MLL binding. This structural change results in the formation of additional electrostatic/polar interactions between KIX and the bound c-Myb, providing a structural basis for the cooperativity observed for the ternary complex. PMID- 16253273 TI - Engineering zinc finger protein transcription factors: the therapeutic relevance of switching endogenous gene expression on or off at command. AB - Modulating gene expression directly at the DNA level represents a novel approach to control cellular processes. In this respect, zinc finger protein DNA-binding domains can be engineered to target virtually any gene. Coupling of a transcription activation or repression domain to these zinc fingers permits regulating gene expression at will, providing a platform of unlimited therapeutic applications. In this review, steps involved in the engineering of zinc finger protein transcription factors are described. In addition, an overview of endogenous genes successfully targeted for modulating expression by engineered zinc finger protein transcription factors is given. So far, research has mainly focused on targeting genes involved in cancer and angiogenesis, with encouraging evaluation in vivo and progression into a clinical trial. Altogether, engineered zinc finger proteins offer a new and exciting direction in the field of medical research with promising prospects. PMID- 16253274 TI - The packaging signal of MLV is an integrated module that mediates intracellular transport of genomic RNAs. AB - Packaging of MLV genomes requires four cis-acting stem-loops. Stem-loops A and B are self-complementary and bind Gag in their dimeric form, while the C and D elements mediate loop-loop interactions that facilitate RNA dimerization. Packaging also requires nuclear export of viral genomes, and their cytoplasmic transport toward the plasma membrane. For MLV, this is mediated by Gag and Env, and occurs on endosomal vesicles. Here, we report that MLV Psi acts at several steps during the transport of genomic RNAs. First, deletion of stem-loop B or C leads to the accumulation of genomic RNAs in the nucleus, suggesting that these elements are involved in export. Second, in chronically infected cells, mutation of the C and D loops impairs endosomal transport. This suggests that RNA dimerization is essential for vesicular transport, consistent with its proposed requirement for Gag binding. Surprisingly, deletion of stem-loop A blocks vesicular transport, whereas removal of stem-loop B has no effects. This suggests that stem-loop A has unique functions in packaging, not predicted from previous in vitro analyses. Finally, in packaging cells that do not express any Psi containing RNA, endosomal RNA transport becomes sequence-independent. This non specific activity of Gag likely promotes packaging of cellular mRNAs. PMID- 16253276 TI - Data mining of tree-based models to analyze freeway accident frequency. AB - INTRODUCTION: Statistical models, such as Poisson or negative binomial regression models, have been employed to analyze vehicle accident frequency for many years. However, these models have their own model assumptions and pre-defined underlying relationship between dependent and independent variables. If these assumptions are violated, the model could lead to erroneous estimation of accident likelihood. Classification and Regression Tree (CART), one of the most widely applied data mining techniques, has been commonly employed in business administration, industry, and engineering. CART does not require any pre-defined underlying relationship between target (dependent) variable and predictors (independent variables) and has been shown to be a powerful tool, particularly for dealing with prediction and classification problems. METHOD: This study collected the 2001-2002 accident data of National Freeway 1 in Taiwan. A CART model and a negative binomial regression model were developed to establish the empirical relationship between traffic accidents and highway geometric variables, traffic characteristics, and environmental factors. RESULTS: The CART findings indicated that the average daily traffic volume and precipitation variables were the key determinants for freeway accident frequencies. By comparing the prediction performance between the CART and the negative binomial regression models, this study demonstrates that CART is a good alternative method for analyzing freeway accident frequencies. IMPACT ON INDUSTRY: By comparing the prediction performance between the CART and the negative binomial regression models, this study demonstrates that CART is a good alternative method for analyzing freeway accident frequencies. PMID- 16253275 TI - Conserved mechanism of Oxa1 insertion into the mitochondrial inner membrane. AB - Oxa1 is the mitochondrial representative of a family of related proteins that mediate the insertion of substrate proteins into the membranes of bacteria, chloroplasts, and mitochondria. Several studies have demonstrated that the bacterial homologue YidC participates both in the direct uptake of proteins from the bacterial cytosol, and in the uptake of nascent proteins from the Sec translocase. Studies on the biogenesis of membrane proteins in mitochondria established that Oxa1 has the capability to receive substrates at the inner surface of the inner membrane. In this study, we asked if Oxa1 may similarly cooperate with a protein translocase within the membrane. Since Oxa1 is involved in its own biogenesis, we used the precursor of Oxa1 as a model protein and investigated its import pathway. We found that immediately after import into mitochondria, Oxa1 initially accumulates at Tim23 that forms the inner membrane protein translocase. Cleavage of the Oxa1 presequence is dependent on mtHsp70, a heat shock protein of the mitochondrial matrix. However, mutant mtHsp70 showing a defect in the release of bound substrate proteins does not interfere with subsequent membrane insertion, indicating that membrane insertion of the mature protein is essentially mtHsp70-independent. We conclude that Oxa1 has the ability to accept preproteins within the membrane. PMID- 16253277 TI - Effect of chronic social stress on nitric oxide synthesis and vascular function in rats with family history of hypertension. AB - Genetic predisposition and psychosocial stress are known risk factors in the aetiology of hypertension. The aim of this study was to investigate the as yet unknown role of nitric oxide (NO) in mechanisms of social stress-induced hypertension in rats with a family history of hypertension. Male adult rats used in the study were offspring of normotensive (Wistar) dams and spontaneously hypertensive sires. The rats were exposed to 6-week crowding stress (5 rats/cage, 200 cm2/rat). Control rats were kept four per cage (480 cm2/rat). Blood pressure was determined non-invasively on the tail. Basal blood pressure of all rats was 131 +/- 2 mm Hg. Crowding stress increased significantly blood pressure (p < 0.02 vs. basal value). Crowding had no influence on NO synthase activity in the left ventricle, adrenal glands and kidney. However, crowding stress reduced significantly NO synthase activity in the aorta by 37% (p < 0.01 vs. control). Acetylcholine-induced relaxation and noradrenaline-induced vasoconstriction of the femoral artery were reduced in stressed rats by 58% (p < 0.001) and 41% (p < 0.003), respectively. On balance then, the results indicate that chronic social stress produced by crowding was associated with reduced vascular NO synthesis and altered vascular function in adult borderline hypertensive rats of normotensive mothers. PMID- 16253278 TI - Effect of ischemic preconditioning on cerebral blood flow after subsequent lethal ischemia in gerbils. AB - Ischemic tolerance, the phenomenon where a sublethal ischemic preconditioning protects the brain against a subsequent lethal ischemia, has been widely studied. Studies have been done on cerebral blood flow levels prior to the lethal ischemia, but the hemodynamic pattern after global ischemia with ischemic preconditioning has not been reported. Sequential changes in regional cerebral blood flow (rCBF) in gerbil hippocampus after 5 min global ischemia with or without 2 min ischemic preconditioning were studied to determine if ischemic preconditioning affects rCBF. Four different treatments were given: (1) sham operated, (2) 2 min ischemia, (3) non-preconditioned, and (4) preconditioned. Groups (1) and (2) (both groups n = 5) were given a 24-h recovery period and the rCBF was measured for baseline values. 24 h after sham-operation (3) and 2 min ischemia (4), gerbils were subjected to 5 min ischemia followed by 1 h, 6 h, 1 day or 7-day reperfusion periods (all groups n = 5). Although no regional difference was observed in the recovery pattern of rCBF, the values of rCBF were significantly higher in the preconditioned group throughout whole brain regions including hippocampus. These results indicate that ischemic preconditioning facilitated the recovery of rCBF after 5 min global ischemia. It needs further study to determine whether the protecting effects of preconditioning relate to the early recovery of rCBF or not. However, our results could be interpreted that the early recovery of rCBF may lead to benefits for cell survival in the CA1 neuron, probably facilitating other protecting mechanisms. PMID- 16253279 TI - Effects of the essential oil from Citrus aurantium L. in experimental anxiety models in mice. AB - Citrus aurantium L. is popularly used to treat anxiety, among other indications suggesting central nervous system action. Previous studies showed anxiolytic effect in the essential oil from peel in mice evaluated on the elevated plus maze [Carvalho-Freitas, M.I.R., Costa, M., 2002. Anxiolytic and sedative effects of extracts and essential oil from Citrus aurantium L. Biological and Pharmaceutical Bulletin 25, 1629-1633.]. In order to better characterize the activity of the essential oil, it was evaluated in two other experimental models: the light-dark box and the marble-burying test, respectively related to generalized anxiety disorder and to obsessive compulsive disorder. Mice were treated acutely by oral route 30 min (single dose) or once a day for 15 days (repeated doses) before experimental procedures. In light-dark box test, single treatment with essential oil augmented the time spent by mice in the light chamber and the number of transitions between the two compartments. There were no observed alterations in the parameters evaluated in light-dark box after repeated treatment. Otherwise, single and repeated treatments with essential oil were able to suppress marble burying behavior. At effective doses in the behavioral tests, mice showed no impairment on rotarod procedure after both single and repeated treatments with essential oil, denoting absence of motor deficit. Results observed in marble burying test, related to obsessive compulsive disorder, appear more consistent than those observed in light-dark box. PMID- 16253280 TI - Different effects of estradiol and various antiestrogens on TNF-alpha-induced changes of biochemical markers for growth and invasion of human breast cancer cells. AB - In the field of estrogen therapy breast cancer risk is one of the most controversially discussed topic. Actually, the as yet largest placebo-controlled study, the Women's Health Initiative, rather showed a risk reduction, in contrast to observational studies. In the present study we have investigated the effect of estradiol on TNF-alpha-induced changes of various markers in human breast cancer cells and compare it with the effect of the antiestrogens tamoxifen and 2 methoxyestradiol. MCF-7 cells were used for the experiments, the incubation time was 96 h. TNF-alpha elicited a 3-4-fold increase of monocyte-attracting protein-1 (MCP-1) and interleukin-8 (IL-8) as compared to the control value, matrix metalloproteinase-9 (MMP-9) and vascular endothelial growth factor (VEGF) were enhanced by 30 to 40%. E2 alone had no effect on MCP-1, slightly reduced the synthesis of MMP-9 and increased VEGF concentrations by about 20%. In combination with TNF, E2 induced a further stimulation of MCP-1, IL-8 and VEGF, whereby the MMP-9 synthesis was not changed. Tamoxifen and the endogenous estradiol metabolite 2-methoxyestradiol seem to be able to partly inhibit the action of TNF alpha and estradiol. Our results suggest that estrogens may slightly increase tumor growth and spreading beyond the effect of chemokines such as TNF-alpha. However, the magnitude of this E2 effect seems to be marginal as compared to the effect of TNF-alpha alone. The risk of recurrence of breast cancer in patients taking hormone therapy after breast cancer may be slightly enhanced by estrogens, but seems mainly to be driven by the potency of still existing tumor cells to secrete chemokines which can stimulate tumor growth and spreading. PMID- 16253281 TI - Antioxidant and antiradical activities of L-carnitine. AB - L-carnitine plays an important regulatory role in the mitochondrial transport of long-chain free fatty acids. In this study, the antioxidant activity of L carnitine was investigated as in vitro. The antioxidant properties of the L carnitine were evaluated by using different antioxidant assays such as 1, 1 diphenyl-2-picryl-hydrazyl free radical (DPPH.) scavenging, total antioxidant activity, reducing power, superoxide anion radical scavenging, hydrogen peroxide scavenging and metal chelating activities. Total antioxidant activity was measured according to ferric thiocyanate method. alpha-tocopherol and trolox, a water-soluble analogue of tocopherol, were used as the reference antioxidant compounds. At the concentrations of 15, 30 and 45 microg/mL, l-carnitine showed 94.6%, 95.4% and 97.1% inhibition on lipid peroxidation of linoleic acid emulsion, respectively. On the other hand, 45 microg/mL of standard antioxidant such as alpha-tocopherol and trolox indicated an inhibition of 88.8% and 86.2% on peroxidation of linoleic acid emulsion, respectively. In addition, L-carnitine had an effective DPPH. scavenging, superoxide anion radical scavenging, hydrogen peroxide scavenging, total reducing power and metal chelating on ferrous ions activities. Also, those various antioxidant activities were compared to alpha tocopherol and trolox as references antioxidants. PMID- 16253282 TI - Hypothalamic 5-HT functional regulation through 5-HT1A and 5-HT2C receptors during pancreatic regeneration. AB - 5-HT receptors are predominantly located in the brain and are involved in pancreatic function and cell proliferation through sympathetic nervous system. The objective of this study was to investigate the role of hypothalamic 5-HT, 5 HT1A and 5-HT2C receptor binding and gene expression in rat model of pancreatic regeneration using 60% pancreatectomy. The pancreatic regeneration was evaluated by 5-HT content, 5-HT1A and 5-HT2C receptor gene expression in the hypothalamus of sham operated, 72 h and 7 days pancreatectomised rats. 5-HT content was quantified by HPLC. 5-HT1A receptor assay was done by using specific agonist [3H]8-OH DPAT. 5-HT2C receptor assay was done by using specific antagonist [3H]mesulergine. The expression of 5-HT1A and 5-HT2C receptor gene was analyzed by RT-PCR. 5-HT content was higher in the hypothalamus of 72 h pancreatectomised rats. 5-HT1A and 5-HT2C receptors were down-regulated in the hypothalamus. RT-PCR analysis revealed decreased 5-HT1A and 5-HT2C receptor mRNA expression. The 5 HT1A and 5-HT2C receptors gene expression in the 7 days pancreatectomised rats reversed to near sham level. This study is the first to identify 5-HT1A and 5 HT2C receptor gene expression in the hypothalamus during pancreatic regeneration in rats. Our results suggest the hypothalamic serotonergic receptor functional regulation during pancreatic regeneration. PMID- 16253283 TI - After effects of noise-induced sleep disturbances on inhibitory functions. AB - The study focuses on possible after effects of noise-induced sleep disturbances on inhibitory brain processes reflecting in performance changes and alternations of inhibition-related components of event-related potentials (ERPs). Following a quiet night and three nights, in which railway noise was presented with different levels, twelve women and ten men (19-28 years) performed a visual Go/Nogo task that contained stimuli either compatible or incompatible with a response. Noise induced sleep disturbances are highly evident in worsening of subjective sleep quality but did not show up in significant changes of reaction time and error rate. A smaller N2 amplitude and longer latency to incompatible than to compatible stimuli as well as an unspecific attenuation of N2 amplitude under Noise were found. The amplitude of the fronto-central P3 was reduced under Noise compared to baseline only in Nogo trials. The amplitude of the parietal P3 in Go trials was smaller to incompatible than to compatible stimuli but was not affected by Noise. Disturbed sleep was associated with a decreased blink rate during task performance. The results suggest that physiological costs to maintain performance are increased after noisy nights. Decisional processes underlying overt responses (Go-P3) are less vulnerable to noise-disturbed sleep than those related to inhibition (Nogo-N2, NoGo-P3). The deficits may have been compensated by increased on-task concentration and thereby did not become apparent in the performance data. Inhibition-related ERPs may be more sensitive indicators of moderate sleep disturbances caused by noise than performance measures. PMID- 16253284 TI - Oxidative stress protection by newly synthesized nitrogen compounds with pharmacological potential. AB - In this study we used new nitrogen compounds obtained by organic synthesis whose structure predicted an antioxidant potential and then an eventual development as molecules of pharmacological interest in diseases involving oxidative stress. The compounds, identified as FMA4, FMA5, FMA7 and FMA8 differ in the presence of hydroxyl groups located in the C-3 and/or C-4 position of a phenolic unit, which is possibly responsible for their free radicals' buffering capacity. Data from the DPPH discoloration method confirm the high antiradical efficiency of the compounds. The results obtained with cellular models (L929 and PC12) show that they are not toxic and really protect from membrane lipid peroxidation induced by the ascorbate-iron oxidant pair. The level of protection correlates with the drug's lipophilic profile and is sometimes superior to trolox and equivalent to that observed for alpha-tocopherol. The compounds FMA4 and FMA7 present also a high protection from cell death evaluated in the presence of a staurosporine apoptotic stimulus. That protection results in a significant reduction of caspase 3 activity induced by staurosporine which by its turn seems to result from a protection observed in the membrane receptor pathway (caspase-8) together with a protection observed in the mitochondrial pathway (caspase-9). Taken together the results obtained with the new compounds, with linear chains, open up perspectives for their use as therapeutical agents, namely as antioxidants and protectors of apoptotic pathways. On the other hand the slight pro-oxidant profile obtained with the cyclic structures suggests a different therapeutic potential that is under current investigation. PMID- 16253285 TI - Inhibition of Naja naja venom hyaluronidase: role in the management of poisonous bite. AB - Hyaluronidase is present virtually in all snake venoms and has been known as a "spreading factor." The enzyme damages the extracellular matrix at the site of the bite, leading to severe morbidity. In this study, the benefits of inhibiting the hyaluronidase activity of Indian cobra (Naja naja) venom have been investigated. Anti-NNH1 and aristolochic acid both inhibited the in vitro activity of the purified hyaluronidase, (NNH1) and the hyaluronidase activity of whole venom in a dose-dependent manner. Both anti-NNH1 and aristolochic acid abolished the degradation of hyaluronan in human skin tissue sections by NNH1 and by whole venom. Aristolochic acid quenched the fluorescent emission of NNH1. A non-competitive mechanism of NNH1 inhibition was observed with aristolochic acid. NNH1 potentiates the toxicity of Daboia russellii VRV-PL-VIII myotoxin and hemorrhagic complex-I. However, the potentiation of toxicity was inhibited dose dependently by anti-NNH1 and aristolochic acid. Further, mice injected with whole venom which had been preincubated with anti-NNH1/aristolochic acid, showed more than a two-fold increase in survival time, compared to mice injected with venom alone. A more moderate increase in survival time was observed when mice were injected with anti-NNH1/aristolochic acid 10 min after whole venom injection. This study illustrates the significance of venom hyaluronidase in the pathophysiology of snake venom poisoning and the therapeutic value of its inhibition. PMID- 16253286 TI - Inhibitory effects of choleretic hydroxyacetophenones on ileal bile acid transport in rats. AB - The effects of the choleretic and cholesterol lowering compound, 2,4,6 trihydroxyacetophenone (THA) and its analog, 2,6-dihydroxyacetophenone (DHA), on ileal bile acid absorption were investigated in rats. THA inhibited taurocholate (TC) uptake into ileal brush-border membrane vesicles (BBMV), showing a maximum inhibition of 50%, whereas DHA completely inhibited TC uptake into ileal BBMV. THA exhibited competitive inhibition with a Ki of 9.88 mM, while DHA showed non competitive inhibition with a Ki of 7.65 mM. Both total and ouabain-sensitive basolateral membrane (BLM) Na+-K+-ATPase activities, which are essential for maintenance of the Na+-gradient for bile acid transport, were inhibited by THA and DHA in a dose-dependent manner. The inhibition of BLM ATPase was uncompetitive with a Ki of 10.1 and 5.0 mM for THA and DHA, respectively. Administration of THA or DHA (400 micromol/kg) twice a day, to hypercholesterolemic rats for 3 weeks caused similar and marked reductions in plasma cholesterol to 60% of the cholesterol-fed controls. The data suggest that the inhibitory actions of THA and DHA on two essential components of ileal bile acid recycling to liver could, in part, contribute to the cholesterol lowering effect of the hydroxyacetophenone compounds. These effects on decreasing bile acid recycling, in combination with their potent choleretic effect, accelerating biliary excretion of bile acids, are responsible for the effective cholesterol lowering capacities of these compounds. PMID- 16253287 TI - Elevated mercury concentrations in failed eggs of Snowy Plovers at Point Reyes National Seashore. PMID- 16253288 TI - An initial economic evaluation of water quality improvements in the Randers Fjord, Denmark. AB - Eutrophication of estuarine and coastal waters has only been recognised as a potential problem since the early 1980s. This problem is being addressed by the European Union through the implementation of recent water quality legislation, which also calls for an economic analysis relating to its implementation. This paper examines the benefits and costs of reduced eutrophication of the Randers Fjord in Denmark, with a primary focus on assessing individual preferences for water quality improvements, using a contingent valuation survey approach. Initial findings of the survey are presented that offer support for funding an action plan to improve the ecological status of the Randers Fjord, as required by the Water Framework Directive. The paper highlights the considerable costs required to abate nutrient emissions into the Fjord, but identifies a need for greater cost evidence at the catchment level. PMID- 16253289 TI - Comparison of mussels and semi-permeable membrane devices as inter-tidal monitors of polycyclic aromatic hydrocarbons at oil spill sites (Boehm et al., 2005). PMID- 16253290 TI - Monitoring the development of intertidal habitats on former agricultural land after the managed realignment of coastal defences at Tollesbury, Essex, UK. AB - The managed realignment of coastal defences and subsequent creation of intertidal habitats is one of several 'soft' engineering options that could reduce the costs of maintaining embankments and at the same time deliver environmental benefits. The managed realignment at Tollesbury was one of the first in the UK, undertaken as an experimental test case to improve understanding of the practical techniques and processes involved. Independent studies were undertaken on the development of soils, benthic invertebrates and vegetation within the site in addition to methods of enhancing the process of natural colonisation of saltmarsh plants. Bathymetric and vegetation monitoring were undertaken on the adjacent estuary to determine any breach effect that may be attributed to the realignment. This paper summarises the results from the vegetation, sedimentation and invertebrate monitoring and discusses the implications for other managed realignment schemes in the UK. PMID- 16253291 TI - Changes in benthic fish assemblages as a consequence of coastal works in a coastal lagoon: The Mar Menor (Spain, Western Mediterranean). AB - The benthic fish assemblage of the Mar Menor consisted of 37 species. Dominant species are: Gobius cobitis, Lipophrys pavo and Tripterygion tripteronotus on infralittoral rocks; Pomatoschistus marmoratus, Callionymus pussillus, Callionymus risso and Solea vulgaris on sandy bottoms and Gobius niger, Syngnathus abaster, Hippocampus ramulosus and Symphodus cinereus on Cymodocea nodosa-Caulerpa prolifera mixed beds. From 1985 to 1989 tourist development has led to the creation of new beaches and the installation of artificial rocky structures for retaining sediments. Dredging for the extraction of sand and subsequent pumping altered sediment characteristics causing a real stress leading to the substitution of typical sandy bottoms communities with Cymodocea nodosa by Caulerpa prolifera communities on mud. Soft bottom fish assemblages responded to changes in vegetation cover and substratum characteristics mainly changing the species composition, while artificial hard substrata contain a similar fish community than natural ones, harbouring even richer and more diverse assemblages. This positive effect of breakwaters should not obscure their likely negative effects on hydrodynamics and the subsequent changes of sediment quality and vegetation cover on the breakwaters' area of influence. PMID- 16253292 TI - Quantification of CD59- mutants in human-hamster hybrid (AL) cells by flow cytometry. AB - Mutation assay is an important approach in evaluating the genotoxic risk of potentially harmful environmental chemicals. The human-hamster hybrid (A(L)) cell mutagenesis system, based on the complement/antibody-mediated cytotoxicity principle, has been used successfully to evaluate the mutagenic potential of a variety of environmental toxicants. The A(L) cells contain a standard set of CHO chromosomes and a single human chromosome 11, which expresses several cell surface proteins including CD59 encoded by the CD59 gene at 11p13.5. A modified mutation assay by flow cytometry was developed to determine the yield of CD59- mutants after either radiation or chemical treatment. After incubation with phycoerythrin-conjugated mouse monoclonal anti-CD59 antibody, the CD59- mutant yields were determined by quantifying the fluorescence of the cells using flow cytometry. This method is faster and eliminates the commonly encountered toxicity problems of the complements with the traditional complement/antibody assay. By comparing the mutant fractions of radiation or chemically treated A(L) cultures using the two methods, we show here that the flow cytometry assay is an excellent substitute in providing an efficient and highly sensitive method in mutant detection for the traditional complement/antibody assay. PMID- 16253293 TI - Bilateral dorsolateral thalamic lesions disrupts conscious recollection. AB - In an earlier study we disputed the claim that the mediodorsal thalamic nucleus is critical for familiarity. We reported patient (QX) who showed a severe deficit in conscious recollection, and behavioural problems (disinhibition, emotional lability) with relative sparing of familiarity-aware memory following a left mediodorsal thalamic lesion. More recent MR imaging has revealed that QX's lesions are more extensive than previously reported and involve both dorsolateral thalamic nuclei, and whilst there is evidence of left mediodorsal thalamic damage, it is not the main focus of damage. This paper reports a full analysis of QX's thalamic pathology alongside a more detailed investigation of his recognition memory, using yes/no and forced-choice procedures, and executive function. The results revealed impairments in yes/no recognition and conscious recollection rates of famous, artist and unknown names. In addition to the previously noted behavioural disinhibition and emotional lability, a deficit in spontaneous planning ability was evident on the Zoo Map Test (subtest of the Bahavioural Assessment of the Dysexecutive Syndrome). Forced-choice recognition, familiarity estimates and remote memory showed higher levels of preservation. The findings indicate that the dorsolateral thalamus is part of the extended hippocampal circuit which is causally critical only for recall and conscious recollection of complex associations rather than for the more automatic processes linked with novelty detection. PMID- 16253294 TI - The role of local and global processing in the recognition of living and nonliving things. AB - We report a study on a patient (DW) with integrative visual agnosia and a category-specific recognition impairment for living things. We assessed DW's local and global processing and tested if his integrative agnosia could have led directly to his category-specific impairment. The main findings were: (i) DW was faster at identifying local compared to global letters. (ii) DW showed no local to-global (or global-to-local) interference effects in selective attention tasks. (iii) DW showed a congruency effect in a divided attention task, suggesting that, when his attention was cued to both levels, he could process information simultaneously and integrate local and global information. (iv) Controls were poorer at naming nonliving compared to living things when presented with silhouettes. These data suggest that local and global information are differentially weighted in the visual recognition of living and nonliving things, and that an impairment in processing the overall shape of an object can lead to a category-specific deficit for living things. Crucially, this implies that category-specific impairments do not necessarily reflect damage to the semantic system, and models of semantic memory based on this assumption need to be revised. PMID- 16253295 TI - Temporal characteristics of feeding behavior in the Munich miniature pig. AB - Correlations between meal size and inter-meal intervals as a characteristic of ingestive behavior are generally calculated utilizing Pearson's correlation coefficient. However, this commonly used method may exhibit disadvantages and methodological problems when dealing with dependent time series. Alternatively, a modified version of Pearson's r (r(diff)) or a nonparametric procedure, Pfanzagl's T combined with an expanded sampling theorem offer a statistical alternative for correlational analysis, robust to stochastic interdependencies between time series. In two studies, porcine self-initiated feeding behavior was observed in a closed economy setup, and a comparison of all three correlation coefficients was performed. STUDY 1: Twelve Munich mini-pigs were observed for 2 weeks. The rate of food ingestion was a stable characteristic of each individual pig, feeding and defecation behavior were temporally associated and light/dark cycle differences were seen for feeding. STUDY 2: Eight Munich mini-pigs were studied for 4 weeks. Kendall's Tau was utilized to test the stability of feeding behavior across weeks. Stability increased over time and feeding was more stable during the light period. Both correlation measures revealed a strong association between meal size and inter-meal intervals. CONCLUSION: Munich-miniature pigs exhibited a stable pattern of self-initiated ingestive and excretory behaviors. In both studies, Pfanzagl's T-values as well as r(diff)-values revealed a corresponding and consistent association between meal size and inter-meal intervals. In combination with the conventional Pearson's r, all three coefficients characterize different aspects of feeding behavior. PMID- 16253296 TI - Sleep deprivation and daily torpor impair object recognition in Djungarian hamsters. AB - Sleep has been shown to play a facilitating role in memory consolidation, whereas sleep deprivation leads to performance impairment both in humans and rodents. The effects of 4-h sleep deprivation on recognition memory were investigated in the Djungarian hamster (Phodopus sungorus). Because sleep during the first hours after daily torpor has many similarities to recovery from sleep deprivation, the effects of spontaneous torpor on object recognition were also assessed. A 4-h sleep deprivation, starting immediately after an object learning task, diminished the ability of the hamsters to: (1) discriminate between an already encountered object (target) and a novel object presented in a novel context, (2) retrieve a target within a complex spatial scene, and (3) detect a spatial rearrangement of familiar objects in a familiar context. Plasma stress hormone levels were similar in sleep-deprived and control hamsters. The occurrence of a daily torpor episode during retention was associated with impaired old-new object discrimination performance in the more effortful complex spatial scene task only, and in a two object choice situation in a novel context no torpor-induced deficit was found. Our results show that post learning sleep deprivation and daily torpor induce a deficit in familiar object retrieval performance in a complex spatial scene, while sparing familiarity-based recognition and novelty processing. Sleep deprivation during the first 4 h of memory consolidation hampered also recency memory for discrete objects. Stress was not a factor contributing to the sleep deprivation-induced impairment. PMID- 16253297 TI - Structural characterization of phytotoxic terpenoids from Cestrum parqui. AB - Isolation, chemical characterization and phytotoxicity of nine polyhydroxylated terpenes (five C13nor-isoprenoids, two sesquiterpenes, a spirostane and a pseudosapogenin) from Cestrum parqui L'Herr are reported. In this work we completed the phytochemical investigation of the terpenic fraction of the plant and described the structural elucidation of polar isoprenoids using NMR methods. All the configurations of the compounds have been assigned by NOESY experiments. Four new structures have been identified as (3S,5R,6R,7E,9R)-5,6,9-trihydroxy-3 isopropyloxy-7-megastigmene, 5alpha-spirostan-3beta,12beta,15alpha-triol, and 26 O-(3'-isopentanoyl)-beta-d-glucopyranosyl-5alpha-furost-20(22)-ene-3beta,26-diol, and as an unusual tricyclic sesquiterpene. The compounds have been assayed for their phytotoxicity on lettuce at the concentrations ranging between 10(-4) and 10(-7)M. The activities of some compounds were similar to that of the herbicide pendimethalin. PMID- 16253298 TI - Quassinoids from the leaves of the Madagascan Simaroubaceae Samadera madagascariensis. AB - An investigation of the leaves of the Madagascan Simaroubaceae Samadera madagascariensis has yielded three C18 quassinoids, 5beta,6-dihydrosamaderine A, 2-chlorosamaderine A, and samaderolactone A, and a C19 quassinoid, 3,4beta dihydrosamaderine C, together with the known quassinoids samaderine A, samaderine B, and cedronin. The compounds isolated displayed little or no anti-tumour activity. PMID- 16253299 TI - Identification of bovine dendritic cell phenotype from bovine peripheral blood. AB - Dendritic cells (DCs) are professional antigen presenting cells, which initiate primary immune responses and also play an important role in the generation of peripheral tolerance. There is no reliable method established for the isolation of bovine peripheral blood DCs, and furthermore, the phenotypes and the functions of bovine DCs are still not fully clear. In the present study, we have attempted to identify bovine peripheral blood DCs by negative-selection. In bovine peripheral blood mononuclear cells (PBMC), we have newly characterized the phenotype of DCs, which is CD11c+/CD172a+. These cells display features of myeloid type DCs. In the thymic medulla, CD11c+/CD172a+ cells were also present and CD1+/CD172a+ cells were additionally detected as a population of DCs. The data suggest that one of the bovine DCs phenotypes from PBMC is derived from myeloid lineages lacking a CD1 molecule, which then drift to several tissues, and that they then may express a CD1 molecule upon their functional differentiation. PMID- 16253300 TI - Rapid assessment for prioritisation of trachoma control at community level in one district of the Kaolack Region, Senegal. AB - The objective of this study was to use a modified Lot Quality Assurance Sampling methodology to classify communities according to prevalence of active trachoma and to estimate the prevalence of trachoma and trichiasis in Nioro department, Kaolack Region, Senegal. A survey was conducted using two-stage cluster sampling to select 50 children aged 2-5 years in each of 33 clusters. In total 1,648 children were examined for active trachoma. Information on trachoma risk factors was collected through interviews with the mother or the household head of the child. Adults (>40 years) with trichiasis were identified through case finding. Nineteen clusters had a low prevalence of active trachoma in children aged 2-5 years (<20%), 11 had medium prevalence (20-40%) and three had high prevalence (>40%). The prevalence of active trachoma in children aged 2-5 years was 17.4% (95% CI 12.9-21.8%). Multivariate-adjusted predictors of active trachoma were: age, facial cleanliness, hygiene practices and keeping cattle in the household. The prevalence of trichiasis in adults aged over 40 years was 1.77% (95% CI 1.24 2.51), equating to 985 adults (95% CI 765-1250) with trichiasis in Nioro department. In conclusion, a survey using rapid methodology showed that trachoma is a problem of public significance in Nioro department, Senegal. PMID- 16253301 TI - Nongenomic effects of estrogen: why all the uncertainty? AB - It is clear that estradiol has profound, rapid effects on the conformation of the estrogen receptors (ERs), ERalpha and ERbeta, which mediate the transcriptional effects of estradiol. Estrogen can elicit many other rapid changes in cells including changes in ion fluxes across membranes and stimulation of kinases and phosphatases. The proteins which are the targets of these actions are the subject of intense investigation. One of the issues that have not been satisfactorily resolved is whether ERalpha or ERbeta can reside in the plasma membrane and participate in the rapid effects of estrogen. In the present commentary, we take a careful look at some of the published data in an attempt to understand why it is so difficult to obtain a definitive answer to this question. PMID- 16253302 TI - HIV-1 Nef stabilizes AP-1 on membranes without inducing ARF1-independent de novo attachment. AB - HIV-1 Nef affects the trafficking of numerous cellular proteins to optimize viral replication and evade host defenses. The adaptor protein (AP) complexes, which form part of the cytoplasmic coat of endosomal vesicles, are key cellular co factors for Nef. Nef binds these complexes and alters their physiologic cycle of attachment and release from membranes. Specifically, while AP-1 normally becomes cytosolic when attachment events are blocked by inhibition of the GTPase cycle of ADP-ribosylation factor-1 (ARF1), the complex remains membrane-associated in Nef expressing cells. To investigate the mechanism of this effect, we used a permeabilized cell system to detect the de novo attachment of exogenous AP-1 to endosomal membranes. Nef did not mediate de novo attachment independently of ARF1, despite its ability to maintain the association of AP-1 with endosomal membranes when the activity of ARF1 was blocked. We conclude that Nef stabilizes AP complexes on endosomal membranes after ARF1-dependent attachment. This stabilization may facilitate coat formation and stimulate the trafficking of multiple cellular proteins. PMID- 16253304 TI - Characterization of phosphorus-releasing bacteria in a small eutrophic shallow lake, Eastern China. AB - Phosphorus contents and phosphorus-releasing bacteria were characterized in a small eutrophic lake, Eastern China. Total phosphorus and water soluble inorganic phosphate (WSIP) in water bodies were as high as the levels of hyper-eutrophic lakes. Calcium bound phosphate and organic phosphorus were 2 major forms of phosphorus existing in the sediments. Enumeration of inorganic phosphate solubilizing bacteria (IPB) and organic phosphorus-mineralizing bacteria (OPB) with culture-dependent method showed that these bacterial groups were not very rich in the ecosystem. Molecular identification and phylogenetic analysis of the predominant IPB and OPB strains indicated that there existed various kinds of bacteria participating in the phosphorus release. Laboratory tests on phosphorus release abilities showed IPB strains could liberate more than 50 microgml(-1), while OPB strains produced less than 2 microgml(-1) WSIP every day. PMID- 16253303 TI - TNF-alpha and IFN-alpha enhance influenza-A-virus-induced chemokine gene expression in human A549 lung epithelial cells. AB - Lung epithelial cells are the primary cellular targets for respiratory virus pathogens such as influenza and parainfluenza viruses. Here, we have analyzed influenza A, influenza B and Sendai virus-induced chemokine response in human A549 lung epithelial cells. Influenza virus infection resulted in low CCL2/MCP-1, CCL5/RANTES, CXCL8/IL-8 and CXCL10/IP-10 production at late times of infection. However, when cells were pretreated with TNF-alpha or IFN-alpha, influenza-A virus-induced chemokine production was greatly enhanced. Cytokine pretreatment resulted in enhanced expression of RIG-I, IKKepsilon, interferon regulatory factor (IRF)1, IRF7 and p50 proteins. Most importantly, influenza-A-virus-induced DNA binding of IRF1, IRF3, IRF7 and NF-kappaB onto CXCL10 ISRE and NF-kappaB elements, respectively, was markedly enhanced in cytokine-pretreated cells. Our results suggest that IFN-alpha and TNF-alpha have a significant role in priming epithelial cells for higher cytokine and chemokine production in influenza A virus infection. PMID- 16253305 TI - Characterization of natural organic matter in conventional water treatment processes for selection of treatment processes focused on DBPs control. AB - Natural organic matter (NOM) from raw and process waters at a conventional water treatment plant was isolated into hydrophobic and hydrophilic fractions by physicochemical fractionation methods to investigate its characteristics. Formation potential of trihalomethanes (THMs) was highly influenced by the hydrophobic fraction, whereas haloacetic acids formation potential (HAAFP) depended more on the hydrophilic fraction. However the hydrophobic fraction was removed more than the hydrophilic fraction through conventional water treatment. Therefore residual hydrophilic NOM after conventional treatment needs to be removed to reduce HAAFP. Feasible additional processes are required to be evaluated by comparing preferential removal efficiency of hydrophilic NOM through pilot tests. The structural and chemical characteristics of hydrophobic NOM (i.e., humic substances (HS)) were further investigated to know how they are influenced by conventional treatment. The phenolic fraction in the hydrophobic NOM was mainly removed compared to the carboxylic fraction through water treatment, and a higher formation potential of THMs resulted from NOM with a higher phenolic content. The Fourier-transform infrared (FT-IR) and proton nuclear magnetic resonance ((1)H-NMR) employed for characterization of NOM through water treatment were insightful revealing that their results were quite close to each other. Decreases of ratio of UV absorbance at 253 and 203 nm, respectively (A(253)/A(203) ratio) and trihalomethane formation potential/dissolved organic carbon (THMFP/DOC) showed consistent trends; therefore, the A(253)/A(203) ratio may be a good indicator of tendency for the formation potential of disinfection by-products (DBPs). PMID- 16253307 TI - Heavy metals effects on forage crops yields and estimation of elements accumulation in plants as affected by soil. AB - Heavy metals (Cu, Cd, Pb, Zn) effect on the productivity of forage crops (clover and perennial cereal grasses) and their accumulation in plants, depending on the concentration of these elements in a soil, has been studied in micro-field experiments on three types of soil. The principle objective was to determine regularities of heavy metals migration in a soil-plant system aiming the estimation of permissible levels of heavy metals content in soils with the following elaboration of methods, which regulate the toxicants transfer to plants. Methods of field experiments, agrochemical and atomic absorption analysis were used. Results were statistically treated by Statistica 6.0, S-Plus 6. Experimental results have shown that the intensity of heavy metals accumulation in plants depends on the type of the soil, the species of plants, the physicochemical properties of heavy metals and their content in the soil. Logarithmic interdependency of heavy metals concentration in soils and their accumulation in plants is suggested. However, the strong correlation between the different heavy metals concentrations in the various soils and the yield of crops was not observed. Toxicants accumulation in crops decreased in time. PMID- 16253306 TI - River water quality of the River Cherwell: an agricultural clay-dominated catchment in the upper Thames Basin, southeastern England. AB - The water quality of the River Cherwell and a tributary of it, the Ray, are described in terms of point and diffuse sources of pollution, for this rural area of the upper Thames Basin. Point sources of pollution dominate at the critical ecological low flow periods of high biological activity. Although the surface geology is predominantly clay, base flow is partly supplied from springs in underlying carbonate-bearing strata, which influences the water quality particularly with regards to calcium and alkalinity. The hydrogeochemistry of the river is outlined and the overall importance of urban point sources even in what would normally be considered to be rural catchments is stressed in relation to the European Unions Water Framework Directive. Issues of phosphorus stripping at sewage treatment works are also considered: such stripping on the Cherwell has reduced phosphorus concentrations by about a factor of two, but this is insufficient for the needs of the Water Framework Directive. PMID- 16253308 TI - Modelling of mercury transport and transformation processes in the Idrijca and Soca river system. AB - In the town of Idrija, Slovenia, the world's second largest mercury mine was active for 500 years and about 37,000 tons of mercury has been lost in the environment. Mercury is still drained from soil, riverbed and floodplains and transported with the Idrijca and Soca Rivers to the Gulf of Trieste. A part of inorganic mercury is methylated either in the river system, or later in the coastal area, and, due to its bioaccumulation and biomagnification represents potential danger to human health. A 1-D aquatic model MeRiMod was used to simulate hydrodynamics and sediment transport in the river system from Idrija to the Soca River mouth. Transport of particle bound and dissolved mercury as well as potential net methylation of mercury in the river system was simulated. The simulation of an observed flood wave with 20-year recurrence period was performed in order to validate the model. Methylation was simulated at lower discharges, as higher methylation rates occur in such conditions. The measurement data and the MeRiMod model were also used to establish a historical mercury mass balance of the Idrijca and Soca Rivers catchment. Sediment core data from the Gulf of Trieste and the measured concentrations from floodplains were used to verify and calibrate the model. Simulations of different high discharges were performed as most of the transport of particulate mercury occurs within flood wave conditions. Compared to the measurements, the results of the model showed an agreement within an order of magnitude, for the transport of total mercury mostly within a factor of 4, and for the methylation within a factor of 5. However, proper trends of the phenomena were obtained by simulations. The combination of modelling and measurements has resulted in some interesting conclusions about the phenomenon of the transport and transformations of mercury in the observed river system. PMID- 16253309 TI - The water quality of the River Thame in the Thames Basin of south/south-eastern England. AB - The water quality of the River Thame, a tributary of the River Thames in the Thames basin, is described in relation to point and diffuse contaminant inputs and runoff from permeable and impermeable bedrock geology with their own characteristic water quality. The data is examined to see if the market town of Aylesbury in the upper part of the catchment influences water quality. Previous studies highlighted the influence of Aylesbury sewage treatment works (STW) on soluble reactive phosphorus (SRP) concentrations in the river before and after phosphorus (P) stripping at the STW. Variations in water quality along the river are described and the study indicates that, apart from SRP, water quality determinants seem to be relatively unaffected by Aylesbury. The Thame water quality is compared with other catchment typologies and it is very similar to that of the main stem of the Thames even though the Thames is mainly Chalk groundwater fed. Differences in water quality largely link to the amount of STW effluent within the rivers and to the endmember compositions of the groundwater and near surface water sources. PMID- 16253310 TI - Urinary mercury in adults in Poland living near a chloralkali plant. AB - We conducted a study within the framework of the interdisciplinary European Mercury Emission from Chloralkali Plants (EMECAP) project to assess exposure to mercury (Hg) and the contribution of Hg emissions from a mercury cell chloralkali plant to urinary mercury (U-Hg) in adults living near the plant. We collected data from questionnaires and first morning urine samples from 75 subjects living near the Tarnow plant in Poland and 100 subjects living in a reference area. Median U-Hg was 0.32 mug/g creatinine (microg/gC) and 0.20 microg/gC, respectively. The median U-Hg was also higher in the amalgam-free subjects living near the plant (0.26 microg/gC) than in the reference group (0.18 microg/gC), but no such association was found in a multivariate analysis. There was a statistically significant positive association between U-Hg and number of teeth with amalgams, a negative association with age and a tendency towards higher U-Hg in female subjects. In the amalgam-free subjects there were statistically significant effects of female sex and fish consumption, and a negative association with age. The additional long-term average air Hg concentration from the plant, based on EMECAP environmental measurements and modelling, was estimated to be 1-3.5 ng/m(3) for the residential study area and should have a very small effect on U-Hg. The other Hg emission sources such as coal combustion facilities located nearby should be taken into account in assessing the overall impact of air Hg on U-Hg in this area. PMID- 16253311 TI - FDA perspectives on supplement use by patients on antithrombotic therapy: dietary supplement regulatory overview. PMID- 16253312 TI - A simple technique to determine thrombopoiesis level using immature platelet fraction (IPF). AB - Immature platelet fraction (IPF) has been measured by fully automated analyzer (XE-2100) as reticulated platelet (RP) which is reflected with thrombopoiesis in bone marrow. IPF value in the healthy volunteers was 3.3% (1.0-10.3) and upper 95% confidential interval (95% CI) of IPF was determined as 7.7%. IPF was significantly high in the patients with idiopathic thrombocytopenic purpura (ITP; 17.4%, 1.2-53.2%) and recovery phase of post-chemotherapy, and significantly low in nadir phase of post-chemotherapy, and within normal range in the patients with ITP in complete remission (CR) and with aplastic anemia (AA). Total count of IPF was significantly low in patients with ITP, AA or post-chemotherapy. Mean platelet volume (MPV) was significantly high in only patients with ITP. IPF 7.7% is best point for highest sensitivity (86.8%) and specificity (92.6%) in diagnosis of ITP and recovery phase of post-chemotherapy. In receiver operating characteristic curve for diagnosis of ITP and recovery phase of post chemotherapy, IPF was significantly more useful than MPV. These results show that IPF reflects the pathology of thrombocytopenic disorders, and that measurement of IPF is useful for the differential diagnosis and analysis of platelet kinetics. PMID- 16253314 TI - Ultrastructural and immunohistochemical analysis of rat uroepithelial cell junctions after partial bladder outlet obstruction and selective COX-2 inhibitor treatment. AB - The present study was undertaken to evaluate alterations in uroepithelial cell junctional complexes in partial bladder outlet obstruction (PBOO) of rat bladders using ultrastructural morphometry and immunohistochemistry, and to determine whether selective COX-2 inhibitors have any effects on these structures. A total of 18 male rats were separated into three groups of six rats each: (1) sham operated animals served as controls; (2) a PBOO group, without further treatment (3) and a group that immediately after PBOO, received treatment for 4 weeks with oral Celecoxib, a selective COX-2 inhibitor. Uroepithelial cell junctions were evaluated using transmission electron microscopy combined with morphometry. Results were also assessed by E-cadherin and alpha-catenin immunohistochemistry. Morphometrical analysis of ultrastructural evaluations revealed that 4 weeks of PBOO caused a significant reduction in the electron density of zonula adherens and zonula occludens junctional complexes. Moreover, some desmosomes located between the deeper cells of the uroepithelium showed signs of disintegration. Selective COX-2 inhibitor treatment during 4 weeks of PBOO showed protective effects on adherens and occludens junctions, as well as on desmosomes. Immunohistochemical analysis of E-cadherin confirmed that the decreased E cadherin immunolabelling in 4 weeks of PBOO was prevented by selective COX-2 inhibitor treatment. Based on ultrastructural morphometrical analysis, we conclude that PBOO alone and in combination with selective COX-2 inhibitors can have considerable effects on uroepithelial cellular junctions. Our findings provide a novel area of investigation regarding the selective use of COX-2 inhibitors following PBOO. PMID- 16253315 TI - American or English spelling in scientific manuscripts? Is it time to ditch the diphthong? PMID- 16253317 TI - Differential effects of simultaneous or sequential administration of paroxetine and WAY-100,635 on ejaculatory behavior. AB - Clinical treatment of depression or anxiety with selective serotonin reuptake inhibitors (SSRIs) often results in delayed ejaculation or anorgasmia. Co treatment with subtype-selective serotonin receptor antagonists may alter the timing of onset of action and potentiate or reduce sexual side effects. Sexual behavior in male Sprague-Dawley rats was examined after acute administration of the SSRI, paroxetine and the serotonin1A antagonist, WAY-100,635. Acute administration of paroxetine alone did not alter male ejaculatory behavior. However, administration of paroxetine plus WAY-100,635 resulted in a significant delay in mounting behavior and increased the time to ejaculation. Simultaneous administration of paroxetine and WAY-100,635 produced a greater delay in initiation of mounting behavior and ejaculation compared to sequential administration of paroxetine followed by WAY-100,635. The differential effect on sexual behavior or addition of specific serotonin receptor antagonists may be relevant for clinical treatment therapies of premature ejaculation. PMID- 16253316 TI - Prolonged topotecan infusion with cisplatin in the first-line treatment of ovarian cancer: an NYGOG and ECOG study. AB - OBJECTIVE: To determine the toxicity and efficacy of combined therapy with cisplatin and prolonged infusion topotecan as front line therapy in women with epithelial ovarian cancer. PATIENTS AND METHODS: Women with previously untreated, measurable and non-measurable epithelial ovarian cancer, stages Ic-IV were eligible. Patients were treated with cisplatin 75 mg/m(2) on day 1, followed by topotecan 0.3 to 0.4 mg/m(2)/day given as a continuous infusion over 14-21 days, every 28 days. Dose levels and duration of infusion were adjusted for toxicity as appropriate. Patients were evaluated for response to treatment and treatment toxicity by standard NYGOG criteria. RESULTS: Sixty patients were enrolled. Among the 20 patients with post-surgical residual disease >2 cm, 80% [95% CI (56.3%, 94.3%)] demonstrated an objective response to therapy. The median progression free survival for all 60 patients enrolled was 19.3 months with a median overall survival of 45.6 months given the median follow-up of 55 months (range 6-81 months). Five year survival is estimated to be 41%. Toxicity was observed in the first four patients treated with topotecan (0.4 mg/m(2)/day x 21 days) and dosing was continued at 0.3 mg/m(2)/day x 14 days thereafter. Of the 56 patients treated at the amended dose level, marrow suppression continued to be dose-limiting, with 86% of patients experiencing grade 3 or 4 neutropenia, 55% experiencing grade 3 or 4 thrombocytopenia and 50% of patients experiencing grade 3 or 4 anemia. Nonetheless, only 11/245 cycles administered were associated with febrile neutropenia and/or infection (8 port-related). Other non-hematologic toxicity was as expected. There were no treatment-related deaths. CONCLUSION: This large, multicenter phase II study of prolonged infusion topotecan in combination with cisplatin demonstrated similar response, time to progression and survival compared with reported results of taxane and platinum combinations. Hematologic toxicity was greater but tolerated. Further studies investigating topotecan in combination with platinum therapy as a first line agent are warranted. PMID- 16253318 TI - Prolonged daily exposure to i.v. cocaine results in tolerance to its stimulant effects. AB - We have previously shown that 1 h, but not 6 h, of daily access to i.v. cocaine induces a sensitized response to i.v. cocaine challenge after 14 days of withdrawal. Here we tried to replicate these results using an i.p. cocaine challenge and adding a group of animals that had 1 h daily access to cocaine, but maintained levels of administration comparable to that of saline animals (i.e. a Coc group). Since addiction-associated neuroadaptations are particularly long lasting, we also tested the response to cocaine challenge after a longer withdrawal period of 60 days. Rats had daily access to i.v. self-administered saline or cocaine for 1 h (Coc1h), or to cocaine for 6 h (Coc6h) over 8 days. Subsequently, after 14 days of withdrawal only Coc animals showed a sensitized locomotor response to cocaine challenge administered i.p. After 60 days of withdrawal, i.p. cocaine failed to produce a sensitized response in Coc1h animals and produced a tolerant response in Coc6h animals. The present data support the notion that 6 h of daily access to cocaine leads to different neuroadaptations than those resulting from 1 h of daily access to the drug. In addition, these data further demonstrate a dissociation between sensitization and addiction to cocaine. PMID- 16253319 TI - Pizotyline effectively attenuates the stimulus effects of N-methyl-3,4 methylenedioxyamphetamine (MDMA). AB - MDMA (N-methyl-3,4-methylenedioxyamphetamine) produces a discriminative stimulus (DS) effect in animals, but attempts to completely block this action with selective neurotransmitter antagonists have not been very successful. Biochemically, MDMA can increase synaptic levels of serotonin, dopamine, and norepinephrine that, conceivably, might interact with multiple populations or subpopulations of neurotransmitter receptors. The present study attempted to antagonize the DS effects of MDMA using the nonselective agents clozapine, cyproheptadine, and pizotyline. An extensive and comparative radioligand binding profile was also obtained for the latter two agents. The purported antagonists were administered in combination with the training dose of MDMA to groups of Sprague-Dawley rats trained to discriminate 1.5 mg/kg of MDMA from saline vehicle in a standard two-lever operant paradigm using a VI-15s schedule of reinforcement. Clozapine was without effect at the doses evaluated, and cyproheptadine only partially attenuated MDMA-appropriate responding. In contrast, pizotyline (AD50=2.5 mg/kg), in combination with the MDMA training dose, resulted in a dose related decrease in percent drug-appropriate responding to saline levels. In a separate group of animals trained to discriminate the structurally-related agent N-methyl-1-(4-methoxyphenyl)-2-aminopropane (PMMA) from vehicle, pretreatment with pizotyline also resulted in a substantial decrease in drug-appropriate responding. The results with cyproheptadine and pizotyline in the binding assays confirmed that these agents display high affinity for multiple subpopulations of serotonergic, dopaminergic, adrenergic, histaminergic, and cholinergic receptors. The overall results of the present investigation indicate that pizotyline, which is clinically available in some countries, might be of clinical utility in the treatment of MDMA overdose. PMID- 16253320 TI - Realizing the promise of IVF in cattle--an overview. AB - The in vitro-produced embryo could play a central role in dairy and beef production systems because of its potential role in genetic selection strategies and crossbreeding schemes, and because it can be integrated into reproductive management strategies for improving pregnancy rates in herds with low fertility. The promise attendant upon use of in vitro-produced embryos is not being fully realized, however. Indeed, there are important technical limitations to their production that reduce the desirability of in vitro-produced embryos because of increased costs, sub-optimal embryonic and fetal survival, and offspring that are occasionally abnormal. Most technical problems associated with in vitro production of embryos can be overcome through research. Among the requirements for a successful research program will be renewed emphasis on conducting embryo transfer trials to determine effects of modifications to embryo production protocols on pregnancy rates and fetal development. Given the promise of in vitro embryo technologies, there is an urgent need for a concerted and sustained investment in research to improve these technologies. Developing a consensus that the study of embryo technology is one of the most important areas of agricultural research should be both an individual and organizational priority. PMID- 16253321 TI - After Dolly--ethical limits to the use of biotechnology on farm animals. AB - The cloning of Dolly the sheep gave rise to a widespread call for limits on interference with life. Until recently, the main limits were technical: what it is possible to do. Now scientists are faced with ethical limits as well: what it is acceptable to do. In this context, we take ethics to involve systematic and rational reflection on moral issues raised in the public sphere. The concerns of the general public are not necessarily valid, but they are the best point of departure if the discussion is to lead to a socially robust framework for setting limits to the use of animal biotechnology. To assess public understanding, we examine two sources of data: Eurobarometer surveys from 1991 to 2002 and a qualitative interview study carried out in Denmark in 2000. Based on these sources, we formulate, and then discuss closely, the following concerns: dangers to human health and the environment, animal welfare, animal integrity, and usefulness. In the final part of the article, it is proposed that a principle of proportionality should be the foundation for socially robust applications of animal biotechnology. Only in cases where the usefulness of the technology can be said to outweigh countervailing moral concerns, as in biomedical research, will applications of animal biotechnology stand up to scrutiny in the public sphere. PMID- 16253322 TI - Ovum pick up and in vitro production in the bovine after use in several generations: a 2005 status. AB - The first In Vitro Produced (IVP) calf was born in 1981 and the non-surgical Ovum Pick Up (OPU) technique for the bovine was adapted from the human in 1987. Since then, considerable research has been aimed at improving both technologies in the bovine. Both OPU and IVP can now be seen as mature technologies. It can be estimated that more than 200,000 IVP calves have been born world wide to date, and when the two technologies are combined they are capable of producing over 50 calves per donor cow per year, albeit with a large variation between donors. Not many new breakthroughs are expected for OPU. For IVP however, automation and miniaturization as well as a greater understanding of the embryo through the application of gene based technologies such as micro-arrays, may provide an in vitro environment that is more in vivo-like than traditional micro drop/well systems. This improved environment should result in higher embryo developmental rates as well as improved quality and welfare of subsequent offspring. The application of OPU/IVP has progressed from treating infertile high genetic multiple ovulation and embryo transfer (MOET) cows in commercial situations to enhancing breeding scheme designs. With the bovine genome being rapidly sequenced and bovine genes for traits of economic interest becoming available in the coming years, OPU/IVP will prove invaluable in rapidly multiplying rare genes or Quantitative Trait Loci (QTL) of high value. In due course, it is anticipated that Marker Assisted Selection or Gene Assisted Selection (MAS/GAS) schemes will be more widely implemented. In addition, OPU, and particularly IVP, provide the basis for more advanced technologies such as cloning and transgenics. This paper is dedicated to celebrate and recognize the significant contributions made by Theo Kruip (1939-2003) to the wide area of bovine OPU and IVP. PMID- 16253323 TI - Tubule orientation and the fatigue strength of human dentin. AB - In this study the influence of tubule orientation on the strength of human dentin under static and cyclic loads was examined. Rectangular beams were sectioned from the coronal dentin of virgin extracted molars (N=83) and then loaded in quasi static 4-point flexure or 4-point flexural fatigue to failure. The flexure strength, energy to fracture and fatigue strength were evaluated for specimens with the dentin tubules aligned parallel (theta=0 degrees ) and perpendicular (theta=90 degrees ) to the plane of maximum normal stress. Results from monotonic loading showed that both the flexural strength and energy to fracture of dentin specimens with theta=0 degrees were significantly greater than those with theta=90 degrees . Furthermore, the apparent endurance strength of dentin with theta=0 degrees (44MPa) was significantly greater than that of the dentin with theta=90 degrees (24MPa). The ratio of apparent endurance strength (for fully reversed loading) to the flexure strength for theta=0 degrees and theta=90 degrees was 0.41 and 0.28, respectively. Although the influence of tubule orientation was most important to mechanical behavior, the flexure strength and energy to fracture also decreased with an increase in tubule density. According to differences in the fatigue strength with tubule orientation, restorative practices promoting large cyclic normal stresses perpendicular to the tubules would be more likely to facilitate fatigue failure in dentin with cyclic loading. PMID- 16253324 TI - The degradative resistance of polyhedral oligomeric silsesquioxane nanocore integrated polyurethanes: an in vitro study. AB - Polymer biostability is one of the critical parameters by which these materials are selected for use as biomedical devices. This is the major rationale for the use of polymers which are highly crystalline and stiff namely expanded polytetrafluoroethylene (ePTFE) and Dacron in particular, as arterial bypass grafts. While this is immaterial in high-flow states, it becomes critically important at lower flows with a greater need for more compliant vessels. Polyurethanes being one of the most compliant polymers known are as such, the natural choice to build such constructs. However, concerns regarding their resistance to degradation have limited their use as vascular prostheses and in order to augment their strength, herein a novel polyhedral oligomeric silsesquioxane integrated poly(carbonate-urea)urethane (POSS-PCU) nanocomposite was synthesised by our group. In the following series of experiments, the POSS PCU nanocomposite samples were exposed to accelerated degradative solutions, in an 'in-house' established model in vitro for up to 70 days before being subjected to infra-red spectroscopy, scanning electron microscopy, stress-strain studies and differential scanning calorimetry. Our results demonstrate that these silsesquioxane nanocores shield the soft segment(s) of the polyurethane, responsible for its compliance and elasticity from all forms of degradation, principally oxidation and hydrolysis. These nanocomposites hence provide an optimal method by which these polymers may be strengthened whilst maintaining their elasticity, making them ideal as vascular prostheses particularly at low flow states. PMID- 16253325 TI - The combined effects of crosslinking and high crystallinity on the microstructural and mechanical properties of ultra high molecular weight polyethylene. AB - Ultra high molecular weight polyethylene (PE) has been used for more than forty years as the bearing surface in total joint replacements. In recent years, there have been numerous advances in processing conditions that have improved the wear resistance of this material. In particular, crosslinking has been shown to dramatically improve the wear behavior of this orthopedic polymer in simulator studies. This benefit to wear resistance, however, is accompanied by a decrease in mechanical properties such as ultimate tensile strength, ductility, toughness and fatigue resistance. This degradation to mechanical properties may have serious implications for devices with high stress concentrations or large cyclic contact stresses. Tailoring microstructure for improved structural performance is essential for implant design. In this work we examined the role of crystallinity and crosslinking on the microstructure and mechanical properties of PE. Crystallinity was increased with a high pressure process and crosslinking was obtained with gamma irradiation. Crystallinity was beneficial to fatigue crack propagation resistance and when coupled with crosslinking a polymer with both wear and fatigue resistance was obtained. PMID- 16253326 TI - Differentiating copper and arsenic toxicity using biochemical biomarkers in Asellus aquaticus and Dreissena polymorpha. AB - Biomarkers of metal exposure are well known, but how a suite of such biomarkers will respond if the metal is also an oxidizing agent or causes oxidative stress is unclear. This study compares the effects of copper and arsenic, two metals with different oxidizing potential, on freshwater invertebrates. Dreissena polymorpha and Asellus aquaticus were exposed to nominal concentrations of copper (100 microg/L) or arsenic (80 microg/L) over 7 days, and physiological stress was examined by measuring metallothionein (MT) induction, thiobarbituric acid reactive substances (TBARS), and Na(+)/K(+)-ATPase activity. Both species showed increased levels of MT during 7-day Cu exposure tests and transient changes in lipid peroxidation (TBARS) which decreased to control levels by day 7. Arsenic had no effect on TBARS and only a transitory effect on MT in D. polymorpha over 7 days, although it initially induced lipid peroxidation in A. aquaticus on day 3. No inhibition of the Na(+)/K(+)-ATPase enzyme was observed for exposed organisms, and baseline values reported here, for A. aquaticus, 1.1 micromol Pi/mg/h, and for D. polymorpha, 0.38 micromol Pi/mg/h, are probably the first reported for these species. PMID- 16253327 TI - Mercury, polybrominated diphenyl ether, organochlorine pesticide, and polychlorinated biphenyl concentrations in fish from lakes along an elevation transect in the French Pyrenees. AB - Concentrations of mercury and persistent organic pollutants in fish were examined in 11 lakes of the French Pyrenees spanning an elevation range of 2 km. All lakes were confined to a relatively small area within a 50-km diameter. Most of the lakes were within the Pyrenees National Park, which is restricted to recreational hiking, angling, and seasonal grazing of livestock, and are not subject to any known point sources of contaminants. Fish collected were mainly of 1 species (Salmo trutta fario), which is stocked regularly in the lakes. With increasing elevation, lake temperatures declined along with electrical conductivity and planktonic chlorophyll a. In contrast, water column nutrients (total phosphorus and total nitrogen) and epilithic periphyton biomass were not correlated with lake elevation. Of the global contaminants measured in fish, mercury, dicholorodiphenyltrichloroethane and its derivatives, and polybrominated diphenyl ethers showed the strongest positive correlation with elevation. Hg levels in some fish exceeded health consumption guidelines in these mountain lakes. Variation in fish contaminant levels was not related to differences in growth rate or to fish trophic position as determined using delta15N stable isotope measurements. This implies that the delivery and/or retention of many of these contaminants increase with lake elevation. PMID- 16253328 TI - Rising rates of depression in today's society: consideration of the roles of effort-based rewards and enhanced resilience in day-to-day functioning. AB - Despite the existence of a vastly improved health care system and a multi-billion dollar antidepressant industry, the rates of depression in the US remain alarmingly high. An exploration of lifestyle changes over the past century suggests that the level of physical activity necessary to provide life's basic resources, referred to as effort-based rewards, has diminished in our industrialized, technologically advanced, service-oriented society. The evolution of the accumbens-striatal-cortical circuitry and its modulating neurochemicals in our ancestors played a significant role in sustaining the continued effort critical for the acquisition of resources such as food, water and shelter; consequently, vast reductions in the degree of physical activity required to obtain necessary resources in today's society likely lead to reduced activation of brain areas essential for reward/pleasure, motivation, problem-solving, and effective coping strategies (i.e. depressive symptomology). Comparative cultural and gender analyses reinforce the significant role of effort-based rewards in mood regulation, suggesting that minimal engagement in such endeavors leads to compromised resilience upon exposure to life's stressful challenges. If physical activity is indeed important in the maintenance of mental health, increased emphasis on behavioral and behavioral/cognitive preventative life strategies, as opposed to an emphasis on psychopharmacological strategies directed at very specific neurochemicals after the onset of depression, should be adopted as protective measures against the onset of depressive symptomology. Thus, strategies that include more global neurobiological activation in the relevant context of directed efforts provide a fresh perspective for depression research. PMID- 16253329 TI - Rituals, stereotypy and compulsive behavior in animals and humans. AB - From a survey of the behavior of animals in the wild, in captivity, under the influence of psychoactive drugs and in a model of obsessive-compulsive disorder (OCD), we identify that the behavioral repertoire invariably includes motor rituals, and that such rituals are performed at a few specific locations/objects in the environment with an orderly transition amongst locations/objects. The concept and parameters of this stable organization of rituals in time and space were used to analyze rituals of OCD patients, compared with control individuals performing the same actions (e.g. car locking). It was found that human rituals also converged to a few places/objects where repetitive acts were performed in a regular order, with the acts in OCD patients overlapping with those of control individuals. Across a very diverse range of animals and conditions, motor rituals are thus characterized by their close linkage to a few environmental locations and the repeated performance of relatively few acts. Such similarity in form may reflect a similarity in the mechanisms that control motor rituals in both animals and humans. PMID- 16253330 TI - Mass spectrometry-based relative quantification of human neutrophil peptides 1, 2, and 3 from biological samples. AB - Human neutrophil peptides (HNPs) are cysteine-rich antimicrobial peptides stored in neutrophils. The similar structure of HNPs -1, -2, and -3 renders them impossible to study individually in biological samples. For the first time, we describe a method of individually identifying the HNPs -1-3 from exudative neutrophils using matrix-assisted laser desorption ionization/time-of-flight (MALDI-TOF) mass spectrometry, and we demonstrate the ability to quantify the relative changes in the peptides found in biological samples. The study includes tracheal aspirates (TA) from infants with respiratory syncytial virus (RSV) illness at intubation for respiratory failure (acute illness) and at extubation (convalescence). In vitro, convalescent and acute illness TAs are labeled with d0 and d3-acrylamides, respectively, and mixed 1:1. TA proteins are separated by one-dimensional gel electrophoresis and then identified by mass spectrometry based peptide mass fingerprinting. The ratio of signal intensities for the isotopically normal (d0-labeled) and heavy (d3-labeled) forms of the peptide reveals the relative increase in each peptide with illness. PMID- 16253331 TI - Psychrobacter bacteraemia in a cirrhotic patient after the consumption of raw geoduck clam. AB - Psychrobacter species rarely cause infections in human. We reported herein a 62 year-old cirrhotic patient who presented with fever and diarrhoea after the consumption of raw geoduck clam. Blood culture grew Psychrobacter phenylpyruvicus which was sensitive to most anti-microbial agents. The patient responded promptly to intravenous antibiotics. This is the first report of human infection with this unusual organism which illustrates the potential health hazards of eating raw geoduck clam in patients with advanced liver disease. PMID- 16253332 TI - Fever of unknown origin: what is remarkable in the elderly in a developing country? AB - OBJECTIVES: To investigate fever of unknown origin (FUO) in 97 patients and compare geriatric and adult population. METHODS: We investigated 97 (22 elderly) patients with FUO using the criteria of Petersdorf and Beeson [Medicine 40 (1961) 1] hospitalized between January 1990 and May 2005 at Hacettepe University Hospital. RESULTS: Infectious diseases were the most common cause in the adult (33.3%) and the elderly (45.5%) patients both. Neoplasms were seen in 18.7; 4.5% and collagen vascular diseases were diagnosed in 9.3; 4.5% of the adults and the elderly respectively. Tuberculosis accounted for 60% of all the infectious causes and empirical anti-tuberculous treatment served as a diagnostic method in 43% of the cases with tuberculosis. Lymphadenopathy was more common among the adults with FUO. A diagnosis could be reached in all the elderly patients with a very high erythrocyte sedimentation rate (ESR>100mm/h). At the end of the hospitalization, 14.7% (11/75) of the adult patients and 13.6% (3/22) of the elderly patients died. CONCLUSION: Geriatric patients with FUO usually have characteristics similar to the adult patients with respect to the hospitalization time, diagnosis, and inpatient mortality. Lymphoid organ hyperplasia might be expected less frequently and very high ESR might be a more reliable indicator of systemic disease in the elderly. Empirical anti-tuberculous treatment plays an important diagnostic role in the developing countries with a higher prevalence of tuberculosis. PMID- 16253333 TI - Systemic inflammatory response syndrome in adult patients with nosocomial bloodstream infection due to Pseudomonas aeruginosa. AB - OBJECTIVES: To evaluate relationships between the inflammatory response, clinical course, and outcome of nosocomial BSI due to Pseudomonas aeruginosa. METHODS: We performed a historical cohort study on 77 adults with P. aeruginosa (Pa) nBSI to define the associated systemic inflammatory response syndrome (SIRS). We examined SIRS scores 2 days prior through 14 days after the first positive blood culture. Imipenem resistant--IRPa (n=20) and susceptible infections--ISPa (n=57) were compared. Variables significant in univariate analysis were entered into a logistic regression model. RESULTS: Seventy-four percent of BSI were ISPa and 26.0% by IRPa. Septic shock occurred in 39.0%. Crude mortality was 48.1%. There was no difference in APACHE II (AP2) scores on days -2, -1 and 0 between the ISPa and IRPa groups. Multivariate analysis revealed that AP2> or =20 at BSI onset (P<0.001) and hematologic failure (P=0.001) independently predicted death. CONCLUSIONS: In patients with P. aeruginosa nBSI, the incidence of septic shock and organ failure is high; patients with IRPa BSI are not more acutely ill prior to infection than those with ISPa BSI and outcome is not significantly different; AP2> or =20 at BSI onset and the development of hematologic failure are independent predictors of death. PMID- 16253334 TI - Prevalence and clinical role of GBV-C infection after cardiac surgery in childhood: a study on 414 patients. AB - OBJECTIVES: GB-virus C (GBV-C) and hepatitis C virus (HCV) share similar modes of transmission. We, therefore, examined the prevalence and clinical role of GBV-C and HCV in patients after cardiac surgery in childhood. METHODS: We analysed blood samples of 414 patients and compared them to 487 controls. Evidence of liver disease and risk factors for infection was analysed. RESULTS: Overall prevalence of GBV-C infection was 22.5% in the patients, compared to 6.2% in the controls (HCV infection 11.3 vs. 0.7%). GBV-C RNA was detected in 8.2% of the patients vs. 3.7% in the controls (HCV RNA in 6 and 0%, respectively). Eleven patients had detectable RNA of GBV-C and HCV. 63.4% of patients infected with GBV C and 46.8% of patients infected with HCV cleared the virus from circulation. GBV C infection was not associated with hepatitis. Liver disease was not more frequent in patients co-infected with HCV and GBV-C. CONCLUSIONS: before 1991 have a substantial risk for HCV and GBV-C infection. However, GBV-C infection seems not to be associated with a liver disease. Co-infection with GBV-C and HCV has no influence on long-term clinical outcome or viral clearance of HCV infection. PMID- 16253335 TI - Measurement of neutrophil membrane CD64 and HLA-Dr in a patient with abdominal sepsis. AB - A patient with abdominal sepsis, had both intra and extracellular bacteria in a blood smear, and high levels of neutrophil membrane CD64 and HLA-Dr. Intracellular bacteria are only observed in the terminal phase of a sepsis. Our patient recovered, suggesting that a high expression of neutrophil CD64 is indicative for a good prognosis. PMID- 16253336 TI - Late-onset eosinophilic chronic meningitis occurring 30 years after Taenia solium infestation in a white Caucasian woman. AB - Unlike solitary parenchymal cysts, chronic meningitis is unusual in patients with neurocysticercosis and may poorly respond to treatment. We report the case of neurocysticercosis characterized by severe headache and chronic eosinophilic meningitis occurring 30 years after infestation with Taenia solium. The patient showed considerable improvement following treatment with albendazole and prednisone. PMID- 16253337 TI - A case of post-Japanese encephalitis with partial hypothalamic dysfunction showing repetitive hyperthermia in summertime. AB - Japanese encephalitis (JE) virus is a mosquito-borne virus belonging to the flavivirus family, including the West Nile and St Louis encephalitis viruses endemic to North America. JE virus is prevalent in East Asian countries and can cause acute lethal encephalitis. Although vaccination programs have decreased the incidence of JE in Japan, the cases that do occur are often fatal or associated with considerable clinical sequelae. We report, for the first time to our knowledge, a patient who had repetitive bouts of hyperthermia in the summertime after recovery from acute JE. An insulin challenge test revealed only marginal increases in the levels of beta-endorphin and growth hormone, indicating partial medial hypothalamic dysfunction. Magnetic resonance imaging showed T2 hyperintensity in both thalamic paraventricular subcortical regions, known to project to the hypothalamic paraventricular nucleus. We thus attributed the episodes of hyperthermia to secondary hypothalamic impairment with thalamic lesions. PMID- 16253338 TI - Carotid compression: investigation of cerebral autoregulative reserve in rats. AB - Easy-to-perform, reversible techniques to analyse cerebral autoregulation are still missing in animal research. The carotid compression technique has been established to investigate dynamic cerebral autoregulation in humans. Adapting the carotid compression technique, we compared data from the new application with that of a classical exsanguination method. Compressing the ipsilateral carotid artery with a non-traumatic clip device for 10s modulated cerebral perfusion pressure. After clip release, the peaking laser-Doppler flow velocity increase over the somatosensory cortex allowed calculation of the transient hyperaemic response ratio (THRR) in relation to baseline. Modulating blood-pressure levels maintenance of cerebral blood-flow velocity was compared with THRR responses. With decreasing blood-pressure levels, the THRR first increased (29+/-16% at 95+/ 10 mmHg to 39+/-13% at 75+/-10 mmHg) before it returned to baseline values at 54+/-10 mmHg (27+/-14%). THRR significantly dropped to 11+/-12% at 34+/-11 mmHg when resting cerebral blood-flow velocity levels also started to decline. Based on the close correlation between blood-flow velocity levels and THRR responses, we have concluded that carotid compression is an alternative technique that can be used to assess cerebral autoregulation in rats. The technique allows less invasive and reversible testing of dynamic autoregulation to be performed, and the technique can easily be applied in conjunction with functional tests to potentially allow deeper insights into cerebral vasoregulative mechanisms. PMID- 16253340 TI - A distributed computing system for multivariate time series analyses of multichannel neurophysiological data. AB - We present a client-server application for the distributed multivariate analysis of time series using standard PCs. We here concentrate on analyses of multichannel EEG/MEG data, but our method can easily be adapted to other time series. Due to the rapid development of new analysis techniques, the focus in the design of our application was not only on computational performance, but also on high flexibility and expandability of both the client and the server programs. For this purpose, the communication between the server and the clients as well as the building of the computational tasks has been realized via the Extensible Markup Language (XML). Running our newly developed method in an asynchronous distributed environment with random availability of remote and heterogeneous resources, we tested the system's performance for a number of different univariate and bivariate analysis techniques. Results indicate that for most of the currently available analysis techniques, calculations can be performed in real time, which, in principle, allows on-line analyses at relatively low cost. PMID- 16253339 TI - Isolation of mitochondria with high respiratory control from primary cultures of neurons and astrocytes using nitrogen cavitation. AB - To study neurons or glia-specific mitochondria one needs to isolate these organelles from primary neuronal or astrocytic cell culture. This work provides novel method for isolation of functional and morphologically intact mitochondria from neurons and astrocytes in cell cultures. In the first step, mitochondria are released from cells by disruption of cell membranes using a nitrogen cavitation technique. This technique is based on rapid decompression of a cell suspension from within a pressure vessel. Mitochondria released from cell bodies are then separated from the rest of cell homogenate by Percoll gradient centrifugation. This is a relatively rapid technique that yields to very well coupled mitochondria that exhibited functional and morphological characteristics comparable to mitochondria isolated from brain tissue using common techniques. This technique thus will allow examination of mitochondria that are exclusively cell specific in origin. PMID- 16253341 TI - Isolation of neuronal substructures and precise neural microdissection using a nanocutting device. AB - We describe a set of microfabricated nanocutting devices with a cutting edge of less than 20 nm radius of curvature that enables high precision microdissection and subcellular isolation of neuronal structures. With these devices, it is possible to isolate functional substructures from neurons in culture such as segments of axons and dendrites, dendritic spines and Nodes of Ranvier. By fine tuning the mechanical compliance of these devices, they can also act as alternatives to costly laser capture microdissection workstations for harvesting specific neuronal populations from tissue sections for analysis. The small size of the device (1 mm2x100 microm) allows convenient insertion into researcher specific experimental set-ups. Its ease of use and possibility for batch fabrication makes this a highly effective and versatile tool for tissue microdissection and the microanalysis of neuronal function. PMID- 16253342 TI - Synthesis, spectra, delivery and potentiometric responses of new styryl dyes with extended spectral ranges. AB - Styryl dyes have been among the most widely used probes for mapping membrane potential changes in excitable cells. However, their utility has been somewhat limited because their excitation wavelengths have been restricted to the 450-550 nm range. Longer wavelength probes can minimize interference from endogenous chromophores and, because of decreased light scattering, improve recording from deep within tissue. In this paper we report on our efforts to develop new potentiometric styryl dyes that have excitation wavelengths ranging above 700 nm and emission spectra out to 900 nm. We have prepared and characterized dyes based on 47 variants of the styryl chromophores. Voltage-dependent spectral changes have been recorded for these dyes in a model lipid bilayer and from lobster nerves. The voltage sensitivities of the fluorescence of many of these new potentiometric indicators are as good as those of the widely used ANEP series of probes. In addition, because some of the dyes are often poorly water soluble, we have developed cyclodextrin complexes of the dyes to serve as efficient delivery vehicles. These dyes promise to enable new experimental paradigms for in vivo imaging of membrane potential. PMID- 16253343 TI - Long versus short oligonucleotide microarrays for the study of gene expression in nonhuman primates. AB - The high degree of sequence similarity between human and nonhuman primate (NHP) genomic DNA suggests that human genome sequence-based DNA microarrays may be used effectively to study gene expression in NHP disease models. In the present study, two distinct commercially available human genome microarray platforms, the Affymetrix HG U133A GeneChip System utilizing Human Genome U133A GeneChips and the Applied Biosystems Expression Array System utilizing the Human Genome Survey Microarray, were used to identify and characterize gene expression changes in the anterior cerebellum of a macaque monkey model of human alcoholism. The Affymetrix microarray consists of eleven short oligonucleotide probe sets for each gene while the Applied Biosystems Microarray uses a single long oligonucleotide per gene. Cross-mapping of probes revealed a total of 11,542 genes that are represented on both microarray platforms. Absolute measures of gene expression ("present calls") from the cerebellum RNA samples were 65-70% (Applied Biosystems Expression Array System) and 27-30% (AffymetrixGeneChip System) among these common gene targets. Analysis of variance (ANOVA; p<0.05; >1.2 fold change; detected on at least 50% of the arrays) indicated 932 and 515 differentially expressed genes for the Applied Biosystems and Affymetrix microarrays, respectively. Significance analysis of microarrays (SAM) identified 255 significant genes at 5% false discovery rate (FDR) for the Applied Biosystems data set and five significant genes at 60% FDR (minimum FDR) for the Affymetrix data set. TaqMan assay-based real-time PCR validation of a number of differentially-expressed genes yielded results that agreed well with the array data in the majority of comparisons. This study demonstrates that human sequence based DNA arrays can be used effectively to detect differential gene expression in an NHP disease model and provides evidence that the use of this long oligonucleotide-based microarray platform may be more suitable for cross-species gene expression studies than a short oligonucleotide-based system. PMID- 16253344 TI - Rapid versus non-rapid cycling as a predictor of response to olanzapine and divalproex sodium for bipolar mania and maintenance of remission: post hoc analyses of 47-week data. AB - BACKGROUND: Rapid cycling in bipolar disorder has been associated with greater morbidity. We examine whether rapid cycling affects treatment response to olanzapine or divalproex in acute mania. METHODS: A post hoc analysis of a 47 week, randomized, double-blind study compared olanzapine (5-20 mg/day) to divalproex sodium (500-2500 mg/day) for bipolar manic or mixed episodes (N=251). Young Mania Rating Scale (YMRS) scores > or = 20 were required for inclusion. Patients were classified at study entry as "rapid cyclers" if they experienced > or = 4 episodes within the last year. A repeated measures analysis of variance was used to analyze YMRS change from baseline. RESULTS: A significant three-way interaction (cycling frequency by medication by visit) was found when modeling change in YMRS total scores. For patients with bipolar I disorder identified as rapid cyclers, mania improvement across the trial did not differ significantly between treatment groups (p=0.181). Among non-rapid cyclers, olanzapine-treated patients had significantly greater YMRS improvement than divalproex-treated patients across the trial (p<0.001) and at most time points. Among olanzapine treated patients, non-rapid cyclers experienced numerically greater YMRS improvement than rapid cyclers throughout the trial; statistically significant differences occurred at weeks 11, 15 and 39. In contrast, among divalproex treated patients, YMRS scores were significantly better in rapid cyclers than non rapid cyclers during the first two study weeks but were comparable thereafter. A similar pattern was seen in Clinical Global Impressions-Mania Severity scores. Hamilton Depression scores in rapid versus non-rapid cycling patients differed at some time points but not over the entire trial and differences by cycling status were not treatment-specific. LIMITATIONS: Apart from the post hoc nature of the analyses, there were high dropout rates in both groups, and cycle frequency was not taken into account. CONCLUSIONS: Rapid cycling patients did less well over long-term treatment than non-rapid cycling patients. Among rapid cycling patients, olanzapine and divalproex appear similarly effective against manic symptoms; however, among non-rapid cycling patients, olanzapine-treated patients experienced superior mania improvement. Olanzapine-treated, non-rapid cyclers experienced greater mania improvement than rapid cyclers. The converse was true of divalproex-treated patients early in treatment. PMID- 16253345 TI - The transdifferentiation potential of limbal fibroblast-like cells. AB - We report the identification and isolation of limbal fibroblast-like cells from adult corneo-limbal tissue possessing self-renewing capacity and multilineage differentiation potential. The cells form cell aggregates or clusters, which express molecular markers, specific for ectoderm, mesoderm and endoderm lineages in vitro. Further, these cells mature into a myriad of cell types including neurons, corneal cells, osteoblasts, chondrocytes, adipocytes, cardiomyocytes, hepatocytes and pancreatic islet cells. Despite originating from a non-embryonic source, they express ESC and other stem cell markers important for maintaining an undifferentiated state. This multipotential capability, relatively easy isolation and high rate of ex vivo proliferation capacity make these cells a promising therapeutic tool. PMID- 16253346 TI - A role of the adrenal gland in stress-induced up-regulation of cytokines in plasma. AB - To reveal a pathway by which psychological/physical stresses influence host defense capability, responses to immobilization stress in mice were investigated, focusing on a multifunctional cytokine, interleukin-18 (IL-18). Immobilization stress induced interleukin-18 accumulation in plasma and in the adrenal gland. Inhibition on ACTH resulted in suppressed levels of IL-18 both in plasma and the adrenal gland. In hemi-adrenalectomized mice, plasma IL-18 levels after stress were lower than in sham-operated mice. This, together with the observation in stressed hemi-adrenalectomized mice that IL-6 levels in plasma were suppressed but up-regulated by recombinant IL-18, showed that the adrenal gland plays a crucial role in stress-related elevation of IL-6 in plasma via IL-18. Adrenal gland is highlighted as an organ connecting the psychological, endocrine, and immune systems. Controlling the secretion of IL-18 from the adrenal gland may serve as a possible preventative means against a stress-related disruption of host defenses. PMID- 16253347 TI - Enhanced FoxP3 expression and Treg cell function in pregnant and estrogen-treated mice. AB - Estrogen (E2) upregulates the FoxP3 gene that marks regulatory CD4+CD25+ T cells (Treg cells). However, E2 also inhibits the ability of antigen presenting cells (APC) to activate T cells. It is possible that these opposing functions might affect the degree of overt suppression during pregnancy and autoimmunity. To evaluate E2 effects on Treg cell function, we quantified FoxP3 levels and Treg suppression in CD4+CD25+ T cells from pregnant and E2-treated mice, and overt Treg suppression in E2- vs. placebo-pretreated mice with autoimmune encephalomyelitis. The data clearly demonstrate that enhanced expression of FoxP3, which occurs in pregnant mice and in mice treated exogenously with E2 pellets, results in a concomitant increase in functional suppression within the CD4+CD25(bright) Treg fraction of splenocytes. The similarities in FoxP3 expression and Treg cell function in E2-treated and pregnant mice implicate E2 as a major contributor for increasing Treg function during pregnancy. Surprisingly, suppression was not enhanced when Treg cells from E2-treated mice were activated with APC and CD4+CD25- responder T cells from the same E2-treated mice, a result consistent with impaired APC activation of Treg cells. In contrast, Treg suppression was strikingly enhanced in combined cell cultures from E2-pretreated mice that were protected from EAE induced with neuroantigen in complete Freund's adjuvant. These results suggest that E2 treatment may have opposing effects on Treg cells vs. APC that both contribute to overt suppression, but such effects are overcome and focused towards enhanced suppression in inflammatory environments produced during pregnancy and EAE. PMID- 16253349 TI - The role of nicotinic acetylcholine receptors in lymphocyte development. AB - The sizes of lymphocyte populations in lymphoid organs of nicotinic acetylcholine receptor knockout and chimera (knockout/wild-type) mice were studied by flow cytometry. The absence of beta2 subunit decreased, while nicotine treatment increased B lymphocyte numbers in the bone marrow. In chimera mice, either beta2 or alpha7 subunits influenced lymphocyte populations in primary lymphoid organs, while in the spleen, only alpha7 receptors were critical. More annexin V-positive B cells were found in the bone marrow of knockout than wild-type animals. We conclude that nicotinic receptors are involved in regulating lymphocyte development and control the B lymphocyte survival. PMID- 16253348 TI - Stress induces the translocation of cutaneous and gastrointestinal microflora to secondary lymphoid organs of C57BL/6 mice. AB - Mammals are colonized by a vast array of bacteria that reside as part of the host's microflora. Despite their enormous levels, these microorganisms tend to be restricted to cutaneous and mucosal surfaces. In the current experiment, only a small percentage of non-stressed mice exhibited detectable levels of bacteria in their inguinal lymph nodes (ILN), spleen, liver, or mesenteric lymph nodes (MLN). However, after experiencing repeated social disruption (SDR), a significant increase in the number of animals having bacteria in their ILN and MLN was found. Since SDR involves fighting in which bite wounds on the skin could provide a portal of entry into the host, it was determined whether experimental wounding (full-thickness skin biopsy), chronic restraint (which is a potent stressor that does not disrupt the skin barrier), or wounding combined with restraint would increase the occurrence of bacteria in secondary lymphoid tissues and liver. Wounding did not significantly increase the prevalence of bacteria in the ILN, MLN, or liver. Interestingly, a larger percentage of restrained and restrained plus wounded mice, in comparison to controls, had bacteria in the ILN, MLN, and liver. Although the stressors increased the number of animals that became colonized, the levels of bacteria in the stressed mice were similar to the levels found in the few non-stressed mice that did become colonized. Our results indicate that psychological components of social stress facilitate the translocation of indigenous bacteria into the host, thus identifying an additional facet through which stressors may impact health. PMID- 16253350 TI - An emerging role for PtdIns(4,5)P2-mediated signalling in human disease. AB - Although an established regulator of many cellular functions, the phosphoinositide phosphatidylinositol (4,5)-bisphosphate [PtdIns(4,5)P2) appears to have evaded the attention of drug-discovery companies. An increasing number of reports have identified potential links between PtdIns(4,5)P2-mediated signalling pathways and the aetiology of many human diseases. Here, we review current knowledge of the regulation and function of PtdIns(4,5)P2 and discuss how aberrant PtdIns(4,5)P2-mediated signalling might contribute to human pathologies such as cardiac failure, bipolar disorder, channelopathies and the genetic disorder Lowe syndrome. PMID- 16253352 TI - Olfactory sensitivity for aliphatic aldehydes in CD-1 mice. AB - Using a conditioning paradigm, the olfactory sensitivity of CD-1 mice for a homologous series of aliphatic aldehydes (n-butanal to n-nonanal) was investigated. With all six odorants, the animals significantly discriminated concentrations below 4 ppb (parts per billion) from the odorless solvent, and with n-butanal, n-heptanal, and n-nonanal the best-scoring animals were even able to detect concentrations below 40 ppt (parts per trillion). The results showed (a) CD-1 mice to have a well-developed olfactory sensitivity for aliphatic aldehydes which for the majority of substances is higher than that of other mammalian species, (b) a limited concentration range of about 1 log unit from threshold to near saturation which is similar to that shown at the single receptor cell level, (c) no significant correlation between perceptibility in terms of olfactory detection thresholds and carbon chain length of the aldehydes tested, and (d) no systematic effect of preexposure to an aldehyde on sensitivity. These findings suggest that CD-1 mice are highly sensitive to non pheromonal odorants that are abundant in plant odors. Further, the results demonstrate that sensitivity for members of a homologous series of substances is not a simple function of vapor pressure as threshold values did not correlate with molecular weight. These threshold data may provide useful information for the choice of adequate stimulus concentrations in electrophysiological or imaging studies of the olfactory system or investigations of the discriminative abilities of the mouse. PMID- 16253351 TI - Activity-driven postsynaptic translocation of CaMKII. AB - Ca2+ influx through the NMDA receptor and subsequent activation of Ca2+/calmodulin-dependent protein kinase II (CaMKII) are crucial for learning and one of its physiological correlates, long-term potentiation (LTP). Ca2+/calmodulin promotes CaMKII binding to several postsynaptic proteins, including the NMDA receptor. These interactions strategically place CaMKII at locations where Ca2+ influx through the NMDA receptor is highest for further activation of CaMKII and for phosphorylation of nearby AMPA receptors and of other proteins that are important for LTP. Ca2+-dependent postsynaptic CaMKII clustering is of specific interest because LTP is synapse specific: only synapses that experience LTP-inducing high-frequency activity exhibit LTP. Ca2+-driven protein binding ensures that CaMKII accumulates only at those synapses undergoing LTP. This selectivity is economical and could contribute to the synapse specificity of LTP because downstream effects of CaMKII will occur mainly at synapses that accumulate CaMKII. In this article, we provide an overview of recent progress in postsynaptic CaMKII anchoring and discuss its implication in synaptic plasticity and the etiology and potential treatments of neurological diseases. PMID- 16253353 TI - Putative midkine family protein up-regulation in Patella caerulea (Mollusca, Gastropoda) exposed to sublethal concentrations of cadmium. AB - A cDNA sequence of a putative midkine (MK) family protein was identified and characterised in the mollusc Patella caerulea. The midkine family consists of two members, midkine and pleiotrophin (PTN), and it is one of the recently discovered cytokines. Our results show that this putative midkine protein is up-regulated in specimens of P. caerulea exposed to sublethal cadmium concentrations (i.e. 0.5 and 1 mg l(-1) Cd) over a 10-day exposure period. Semiquantitative RT-PCR and quantitative Real time RT-PCR estimations indicate elevated expression of midkine mRNA in exposed specimens compared to controls. Moreover, RT-PCR Real time values were higher in the viscera (here defined as the part of the soft tissue including digestive gland plus gills) than in the foot (i.e. foot plus head plus heart) of the limpets. At present, information on the functional signalling significance of the midkine family proteins suggests that the up-regulation of P. caerulea putative midkine family protein is a distress signal likely with informative value on health status of the organism and with potential prognostic capability. PMID- 16253354 TI - RNA interference in Haemonchus contortus: suppression of beta-tubulin gene expression in L3, L4 and adult worms in vitro. AB - We have used RNAi to target two beta-tubulin genes in the parasitic stages of Haemonchus contortus in vitro. Soaking exsheathed-L3, L4 and adult worms in medium containing dsRNA resulted in a significant decrease (greater than 1000 fold in some cases) in the expression of the specific beta-tubulin transcript, as measured by quantitative PCR. During the initial 24h exposure to the dsRNA, the gene suppression effect was quite specific to the targeted gene. Six days after initial exposure to dsRNA, treated L3 worms were less able to migrate through a filter mesh, indicating decreased motility, and showed less development to the L4 stage than control larvae. The gene suppression effect occurs in exsheathed L3 larvae despite the fact that this life stage does not have functioning mouthparts, indicating that uptake of dsRNA does not depend on its ingestion. Suppression occurred with dsRNA presented in 'naked' or liposome-encapsulated forms, indicating that a liposome formulation was not necessary for uptake to occur. Adult worms also showed significant gene suppression, however, they did not show any reduced motility compared to controls over a 3-day period. Adult worms treated with ivermectin to paralyse their pharynx, still showed significant gene suppression, again suggesting that uptake of dsRNA does not require ingestion. We have shown that soaking in dsRNA is an effective method for RNAi with the parasitic stages of H. contortus, and, hence, may offer significant potential as a tool for studying gene function in this parasite species. PMID- 16253355 TI - Poor in-hospital outcome in young women with acute myocardial infarction. Does metabolic syndrome play a role? PMID- 16253356 TI - Patient knowledge and perceptions of atrial fibrillation and anticoagulant therapy: effects of an educational intervention programme. The West Birmingham Atrial Fibrillation Project. AB - OBJECTIVE: Previous research has demonstrated that patients with atrial fibrillation (AF) possess very little knowledge of their disease, the consequences of AF and the benefits/risks of anti-coagulant therapy. The aim of this pilot study was to examine patient's knowledge and perceptions of AF and their anticoagulant treatment before and after a brief educational intervention. MATERIALS AND METHODS: Ninety-three patients (47 male; mean (S.D.) age 67.3 (11.6) years) completed the baseline interview to assess their knowledge and perceptions of AF. All patients were given an information booklet which explained what AF was, associated symptoms, the possible causes and consequences of AF, treatment options and their benefits/risks, what the INR is and what factors may effect it. The booklet was explained to each patient by the researcher and given to the patient to take away for reference. Thirty-three (35.5%) patients (12 men; mean (S.D.) age 68.0 (13.6) years) completed the follow-up assessment to re assess their knowledge and perceptions of AF. RESULTS: Only 49% of patients could name their cardiac condition at baseline, although the majority were aware that AF was an arrhythmia (80% baseline vs. 91% follow-up). Only about half the patients perceived AF as a serious condition or were aware that AF predisposes to thromboembolism at baseline. Following the educational intervention there was a non-significant increase in patient knowledge of the risks associated with AF. Of those who completed both questionnaires, 52% were aware that anticoagulants prevented blood clots, which increased to 70% post-intervention. However, few patients were aware of the benefit of stroke prevention associated with anticoagulants at baseline (21%) or after the educational intervention (27%). The intervention had little effect on increasing awareness of the bleeding risks associated with anticoagulants, although three in five people appeared to appreciate these risks. However, the educational intervention significantly improved patient's knowledge of the target INR range and factors that may affect INR levels (p=0.001 and p=0.014, respectively) for those who completed both questionnaires. CONCLUSIONS: Most patients with AF possess very limited knowledge about their cardiac condition, its consequences, and how anticoagulant treatment can benefit them. This pilot study has demonstrated that a brief educational intervention with an information booklet can help to somewhat improve their knowledge about anticoagulation therapy for AF. PMID- 16253357 TI - Thoracic aorta units: an opened avenue to cardiologists. PMID- 16253358 TI - Hydrochlorothiazide induced pulmonary edema--a rare side effect of common diuretic drug. PMID- 16253359 TI - Cardiac magnetic resonance in outpatients in Germany--indications, complications and protocol suggestions from a high-volume center. AB - BACKGROUND: Cardiac magnetic resonance (CMR) has developed into a routine examination in many centers in cardiology. However, there is little knowledge about its applicability in outpatients as a diagnostic tool for cardiovascular diseases. We report about the experiences in a high-volume cardiac imaging center and in a "mobile setting" in Germany and provide routinely used examination protocols. METHODS: 8976 patients referred for CMR from cardiologists, internal medicine practices and from general practitioners and 2200 patients examined in a "mobile" system by outpatient cardiologists were included in the study. Indications were as follows: 7672 (69%) examinations for myocardial ischemia and viability, 1313 (12%) for cardiac and pericardial inflammatory disease and cardiac mass, 976 (9%) for detection and quantification of heart valve disease and 466 (4%) for congenital heart disease. 697 (6%) were referred for other indication. Two independent readers performed image analysis of the 8976 patients in our center. RESULTS: Image quality was rated "excellent" in 90.6%, "good" in 8%, "fair" in 1.2% and "poor" in 0.2%. 0.0002% of all examinations were not assessable due to low image quality. Minor complications (temporarily, asymptomatic AV-blockade; mild chest pain and/or dyspnea; nausea) could be observed in 12% and resolved within few minutes. One patient experienced a grand mal seizure due to hyperventilation. 0.9% examinations had to be terminated untimely due to claustrophobia. CONCLUSION: CMR in outpatients is a widely used imaging modality in cardiology in Germany. A large variety of clinical questions may be answered by CMR with excellent image quality and without major complication. With user-adapted protocols, a rapid diagnosis is achieved even in outpatients in a "mobile" setting. Hence, CMR will increase its applicability as a routine imaging tool. PMID- 16253360 TI - Detection of bovine viral diarrhea virus (BVDV) in seropositive cattle. AB - Detection of bovine virus diarrhoea virus (BVDV) in one vaccinated beef cattle and three non-vaccinated dairy herds was investigated on peripheral blood leukocytes (PBL) with or without previous treatment followed by a capture ELISA (cELISA). Using the combination of PHA and polycation treatment, PBL from 229 seropositive cattle were studied and could be classified in four different states of BVDV infection. Lysed PBL from four animals were directly positive in cELISA (Category I), PBL of 17 animals were positive after PHA stimulation (Category II), 15 animals were positive only after PHA stimulation plus polycation treatment (Category III), while virus could not be detected in 193 seropositive cattle. Wild-type BVDV strains were isolated by co-culture on polycation-treated MDBK cells from 11 of these seropositive animals. BVDV antibodies of these same animals were able to neutralize their own virus, indicating that virus persists in PBL in spite of strain-specific antibodies. No apparent change of leukocyte subpopulations could be detected in any category of virus-positive animals. Thus, BVDV may be present in the PBL of some cattle, even in the presence of a specific active immune response. PMID- 16253361 TI - Evaluation of MAT, IgM ELISA and PCR methods for the diagnosis of human leptospirosis. AB - Serum samples from 125 patients with clinical suspicion of leptospirosis were tested by the microscopic agglutination test (MAT), IgM ELISA and PCR for the diagnosis of the disease. Most patients were adult males and 74.1% (p<0.001) of the patients exposed to water and 77.6% (p<0.001) of those exposed to animals, were respectively considered confirmed or probable cases by MAT. The clinical symptoms mainly observed among the patients considered confirmed or probable cases were fever (95.6%), jaundice and headache (79.4%), myalgia (77.9%), nausea and vomiting (64.7%). About 63% of the confirmed or probable cases were patients that lived in Belo Horizonte, a big city of the Minas Gerais state, Brazil, showing the occurrence of urban leptospirosis. Among the 47 confirmed cases of leptospirosis diagnosed by MAT, 44 (94%) serum samples were positive by IgM ELISA and 17 (36%) were PCR positive. Among the 33 probable cases, 10 (30%) samples showed positive amplification by PCR. By considering MAT as the standard test, the sensitivity and specificity of IgM ELISA was 96.6% and 93.3%, respectively. A relevant finding in our study was the number of positive cases verified by PCR (13-29%) and IgM ELISA (3-7%) among the 45 unconfirmed cases by MAT, demonstrating the value of PCR in the early diagnosis of human leptospirosis. PMID- 16253362 TI - White biotechnology: differences in US and EU approaches? AB - Several predominantly political movements advocate white, or industrial, biotechnology as a means to alleviate economic, ecological and societal problems in petroleum-dependent industrialized nations worldwide. US and European approaches differ significantly and we believe that, in the long-term, only economic drivers will be able to bring about the broad use of renewable resources and a bio-based economy. As long as the cost of fossil fuel and feedstock for key chemicals have not passed their respective critical thresholds, industrial biotechnology and its products will need political support and funding, particularly in the energy and bulk-chemicals sectors. Other uses of industrial biotechnology, however, such as biocatalytic conversions of fine and specialty chemicals and the manufacture of high-value products, such as nutriceuticals, cosmeceuticals and performance chemicals offer dynamic growth opportunities both for established chemical industries, as well as emerging entrepreneurial enterprises. PMID- 16253363 TI - P3a from visual stimuli: task difficulty effects. AB - The P3a event-related brain potential (ERP) was elicited using a visual three stimulus oddball paradigm (target, standard, distracter) in which participants responded only to the target. Discrimination task difficulty between the target and the standard was manipulated by varying the size of the standard stimulus circle relative to a constant target stimulus circle across three conditions (easy, medium, hard). A large checkerboard pattern was employed for the distracter stimulus across all tasks. Error rate and response time increased with increases in task difficulty, so that the task difficulty manipulation was successful. Distracter P3a amplitude increased and target P3b decreased somewhat with increases in task difficulty. The findings suggest that increased perceptual discrimination difficulty between the target and standard stimuli increases P3a amplitude. Theoretical implications are discussed. PMID- 16253364 TI - Changes in plasma volume associated with mental stress ischemia in patients with coronary artery disease. AB - Psychological stress has been shown to trigger angina and myocardial ischemia in patients with coronary artery disease. However, the mechanisms by which stress may trigger cardiac events has yet to be fully elucidated. Twenty five patients underwent radionuclide ventriculography during a multiple stress challenge. Plasma volume was assessed during rest and at the end of the stress task. Flow mediated dilatation was also measured. Controlling for endothelial function and medications, patients with ischemia had greater reductions in plasma volume than non-ischemic patients. Reduced plasma volume may be one mechanism by which mental stress may increase the risk for acute coronary events. PMID- 16253365 TI - Intracellular physiological events of yeast Rhodotorula glutinis during storage at +4 degrees C. AB - Samples of the cheese yeast Rhodotorula glutinis were analysed during storage at +4 degrees C for cultivability, viability, vitality (metabolic activity), membrane potential state, intracellular pH, and carbohydrate content. The results have allowed to describe cellular events occurring during storage. The loss of vitality came with the decrease of carbohydrate content. The fall of trehalose content under a threshold value induced the deterioration of the membrane potential. Later, when all the cells were depolarised, the intracellular pH decreased and the cultivability dropped, whereas viable cells still decreased slowly. Then, it led to an intermediate physiological state similar to the viable but non-cultivable state. Finally, the fall of viability dropped. In this work, we have defined rapid methods relevant to describe the sequence of intracellular events in the cheese yeast R. glutinis during storage, and we applied them to understand the weak vitality without fall of viability of yeast samples. These methods might allow to rapidly test yeast sample quality before use and to predict, at the moment of the harvesting, the conservation of the yeast. PMID- 16253366 TI - The survival of Escherichia coli O157 on a range of metal surfaces. AB - Escherichia coli O157:H7 is a serious pathogen causing haemorrhagic colitis. It has been responsible for several large-scale outbreaks in recent years. E. coli O157:H7 is able to survive in a range of environments, under various conditions. The risk of infection from contaminated surfaces is recognised, especially due to the low infectious dose required. In this study, a high concentration (10(7) cells) of E. coli O157 was placed onto different metals and survival time measured. Results showed E. coli O157 to survive for over 28 days at both refrigeration and room temperatures on stainless steel. Copper, in contrast, has strong antibacterial properties (no bacteria can be recovered after only 90 min exposure at 20 degrees C, increasing to 270 min at 4 degrees C) but its poor corrosion resistance and durability make it unsuitable for use as a surface material. Other copper-containing alloys, such as copper nickels and copper silvers, have improved durability and anticorrosion properties and greatly reduce bacterial survival times at these two temperatures (after 120 min at 20 degrees C and 360 min at 4 degrees C, no E. coli could be detected on a copper nickel with a 73% copper content). Use of a surface material with antibacterial properties could aid in preventing cross-contamination events in food processing and domestic environments, if standard hygiene measures fail. PMID- 16253367 TI - The comparative ability of four isolates of Bacillus subtilis to ferment soybeans into dawadawa. AB - In an attempt to develop starter cultures for fermenting soybeans into the traditional West African condiment dawadawa, four isolates of Bacillus subtilis: 24BP(2), 72RP(17), 72BP(30), and FpdBP(2), which had been selected from 42 Bacillus cultures in a previous study by the current authors, were used separately to produce soy-dawadawa. The accompanying microbiological and biochemical changes, including enzymatic activities, as well as the organoleptic quality of the products were evaluated including that of a control sample which was fermented spontaneously. Significant differences existed in the ability of the four isolates to hydrolyse the soybean proteins, starch, and fat to produce dawadawa. Bacillus subtilis 24BP(2) recorded the highest protease and amylolytic activities, 101 U/ml and 26.68 mg/ml, respectively, and liberated the most amino acids, 117.64 mg/g dry wt., during fermentation. Bacillus subtilis 24BP(2) also grew to the highest population of cells in the final product. Taste panelists found soybean dawadawa produced by each of the four isolates acceptable and rated soup flavoured with soy-dawadawa produced by Bacillus subtilis FpdBP(2) as the best sample. Panelists scored it higher than the control sample and soy-dawadawa produced by Bacillus subtilis 24BP(2) in that order. PMID- 16253368 TI - Adaptation of Chinese hamster ovary cells to low culture temperature: cell growth and recombinant protein production. AB - Recombinant Chinese hamster ovary (rCHO) cells producing erythropoietin (EPO) and rCHO cells producing follicle-stimulating hormone (FSH) showed a significant increase in specific productivity (q) when grown at 32 degrees C compared to 37 degrees C. However, low culture temperature suppressed cell growth, and therefore, did not increase volumetric productivity as much as q. In an attempt to increase the volumetric productivity through improvement of hypothermic growth, EPO producing rCHO (CHO-EPO) cells and FSH producing rCHO (CHO-FSH) cells were adapted at 32 degrees C in a repeated batch mode using spinner flasks. Cell growth of both CHO-EPO and CHO-FSH gradually improved during adaptation at 32 degrees C. Specific growth rates of CHO-EPO and CHO-FSH cells at 32 degrees C, through adaptation, were increased by 73% and 20%, respectively. During adaptation at 32 degrees C, mRNA levels of cold-inducible RNA-binding protein (CIRP) of both rCHO cell lines did not change significantly, suggesting that CIRP expression may not be the only cause for growth suppression at low culture temperature. Unlike cell growth, the recombinant protein production of both rCHO cell lines was not increased during adaptation due to decreased specific productivities. The specific EPO productivity and specific FSH productivity were decreased by 49% and 22%, respectively. Southern blot analyses showed that the decreased specific productivities were not due to the loss of foreign gene copies. Taken together, improvement of hypothermic cell growth by adaptation does not appear to be applicable for enhanced recombinant protein production, since specific productivity decreases during adaptation to the low culture temperature. PMID- 16253369 TI - Coexpression of TorD enhances the transport of GFP via the TAT pathway. AB - Twin-arginine translocation (Tat) pathway is capable of secreting fully folded proteins into the periplasm of Gram-negative bacteria and may thus be an ideal system for the expression of active cofactor-containing proteins. However, the applications of Tat system for such purpose have been plagued by low translocation efficiencies. In this study, we demonstrate that the coexpression of a soluble chaperone, TorD, in conjunction with the TorA signal peptide, the translocation efficiency of GFP can be enhanced by more than three-fold. The enhancement in translocation efficiency is believed to be a result of reduced proteolysis mediated by the binding of TorD toward the TorA signal peptide. We believe this approach can be further exploited for the expression and secretion of other heterologous proteins as well as traditional Tat substrate proteins. PMID- 16253370 TI - Synthesis of polyhydroxyalkanoates from different short-chain fatty acids by mixed cultures submitted to aerobic dynamic feeding. AB - The production of polyhydroxyalkanoates from acetate and propionate by two mixed cultures well adapted to each of these substrates was evaluated. Sludge fed with acetate (A), produced a homopolymer of hydroxybutyrate (HB), whereas sludge fed with propionate (P) produced a copolymer of HB and HV (hydoxyvalerate). Switching the substrate feeds, propionate to sludge A and acetate to culture P, a terpolymer of HB, HV and hydroxymethylvalerate (HMV) was obtained with culture A and a copolymer of P(HB/HV) by sludge P. Regardless of the population used, the polymer yield and productivity were much higher for acetate than for propionate. Feeding a mixture of acetate and propionate, in equal parts, to both cultures resulted in an increase of HV units produced per C mol of propionate consumed, relative to the situation where only propionate was used. The individual use of butyrate and valerate by culture A was also studied. Butyrate produced a homopolymer whereas valerate was stored as a terpolymer of P(HB/HV/HMV). The polymer yields on acetate and butyrate were higher than those on propionate and valerate. The polymer productivity was higher for acetate and propionate than for butyrate and valerate. Results showed that the polymer composition, and consequently the polymer properties, could be manipulated by varying the volatile fatty acid feed composition and/or the population. PMID- 16253371 TI - Real-time monitoring and automatic density control of large-scale microalgal cultures using near infrared (NIR) optical density sensors. AB - Signals from near infrared (NIR) light transmittance sensors were used for both real-time monitoring of algal biomass density in growing mass cultures (200l tubular biofences), and also as feedback in a system that controlled the density of the culture by automatic injection of fresh growth medium. When operated in a semi-continuous production mode between predefined density values, diurnal growth patterns were recorded on-line that provided information on the dynamics of the microalgal cultures with respect to environmental conditions. The bioreactor system was also programmed to operate in constant biomass density mode, thereby maintaining the culture at the optimal population density (OPD), and sustaining high biomass production levels. The system has potential for operating a dynamic density set point for microalgal cultures where the optimal population density varies as a function of ambient growing conditions. PMID- 16253372 TI - Enzymatic recovery and purification of polyhydroxybutyrate produced by Ralstonia eutropha. AB - Polyhydroxybutyrate (PHB) is the most studied among a wide variety of polyhydroxyalkanoates, bacterial biodegradable polymers known as potential substitutes for conventional plastics. This work aimed at evaluating the use of enzymes to recover and purify the PHB produced by Ralstonia eutropha DSM545. Screening experiments allowed the selection of trypsin, bromelain and lysozyme among six enzymes, based on their efficiency in lysing cells of a non-PHB producing R. eutropha strain. Then, process conditions for high efficiency in PHB purification from the DSM545 cells were searched for the enzymes previously selected. The best result was achieved with 2.0% of bromelain (enzyme mass per biomass), equivalent to 14.1 U ml(-1), at 50 degrees C and pH 9.0, resulting in 88.8% PHB purity. Aiming at improving the process efficiency and reducing the enzyme cost, experiments were carried out with pancreatin, leading to 90.0% polymer purity and an enzyme cost three times lower than the one obtained with bromelain. The molecular mass analysis of PHB showed no polymer degradation. Therefore, this work demonstrates the potential of using enzymes in order to recover and purify PHB and bacterial biopolymers in general. PMID- 16253373 TI - Cloning of a functional Salmonella SPI-1 type III secretion system and development of a method to create mutations and epitope fusions in the cloned genes. AB - Bacterial type III secretion systems have significant potential to be harnessed for beneficial purposes including vaccine development, anti-cancer therapies, strategies to counteract harmful bacteria-host interactions, and evolutionary studies. The ability to clone and manipulate type III secretion systems would allow researchers to perform novel experiments that would progress the biotechnological development of the potentially positive uses of these systems. Here, we report the cloning of the entire Salmonella pathogenicity island 1 (SPI 1) type III secretion system on a single DNA fragment that is contained on a self transmissible plasmid vector for convenient transfer to alternate hosts. We demonstrate that the cloned SPI-1 type III system is functional for secretion and translocation via complementation of an S. typhimurium Delta SPI-1 strain. We also present a convenient method to construct mutations and epitope fusions in the cloned type III genes and demonstrate that the engineered substrate protein fusions are recognized by the cloned type III system. We transferred the cloned SPI-1 type III system into bacterial strains of different genera and found that there is a SPI-1 gene expression defect in these strains. The results describe a novel strategy for cloning and manipulation of bacterial secretion system gene clusters and provide a foundation for future studies to develop the beneficial uses of cloned type III secretion systems. PMID- 16253374 TI - Nineteen nucleotides in the variable region of 3' non-translated region are dispensable for the replication of dengue type 1 virus in vitro. AB - In many flaviruses, first 50-400 nucleotides of 3' non-translated region (3' NTR) exhibit lower conservation level than other regions and are called "variable region". Two dengue type 1 virus (DENV-1) strains, which have 17- and 29-nt deletion in the variable region, were recently isolated from Japanese dengue fever patients. The effect of a small deletion in the 3' NTR was analyzed using two DENV-1 viruses which were prepared from a newly developed infectious cDNA clone. These included a recombinant virus rDENV-1(02-20), without any deletion in 3' NTR, and rDENV-1m10, with 19-nt deletion in the variable region of rDENV-1(02 20). These two viruses were compared for growth kinetics and plaque morphology in Vero, Huh-7 and C6/36 cells. No apparent difference was detected between rDENV 1(02-20) and rDENV-1m10 in replication efficiency and plaque size in these cell lines. The results suggest that the complete variable region of DENV-1 is dispensable for virus replication and propagation in vitro. PMID- 16253375 TI - Mobility of model proteins in hydrogels composed of oppositely charged dextran microspheres studied by protein release and fluorescence recovery after photobleaching. AB - In this paper, the release of proteins from a novel self-gelling hydrogel based on biodegradable dextran microspheres is investigated. The protein-loaded macroscopic gels are obtained by hydration of mixtures of oppositely charged hydroxyethyl methacrylate-derivatized dextran microspheres with a protein solution. In media of low ionic strength (100 mM Hepes pH 7.0) it was found that the release of the entrapped model proteins (lysozyme, BSA and IgG) was slower than in saline (150 mM NaCl, 100 mM Hepes pH 7.0). The reason behind this observation is that substantial adsorption of the proteins onto the microspheres' surface and/or absorption in the microspheres takes place. Confocal images showed that independent of their crosslink density the microspheres are impermeable for BSA and IgG. BSA, bearing a negative charge at neutral pH, was adsorbed onto the surface of positively charged microspheres. Lysozyme, which is positively charged at neutral pH, was able to penetrate into the negatively charged microspheres. In saline, the gels showed continuous release of the different proteins for 25 to 60 days. Importantly, lysozyme was quantitatively and with full preservation of its enzymatic activity released in about 25 days. This emphasizes the protein friendly technology to prepare the protein-loaded gels. Mathematical modeling revealed that protein release followed Fick's second law, indicating that the systems are primarily diffusion controlled. These results show that these hydrogels are very suitable as injectable matrix for diffusion-controlled delivery of pharmaceutically active proteins. PMID- 16253376 TI - Asialoglycoprotein receptor targeted gene delivery using galactosylated polyethylenimine-graft-poly(ethylene glycol): in vitro and in vivo studies. AB - The asialoglycoprotein receptor (ASGP-R) on the hepatocyte membrane is a specific targeting marker for gene and drug delivery. Polyethylenimine (PEI) is a polycationic nonviral vector that is used for gene transfer. We have synthesized galactosylated polyethylenimine-graft-poly(ethylene glycol) (GPP) for performing gene delivery to the hepatocytes. The present study reports on the in vitro and in vivo data that was achieved in hepatoma bearing transgenic mice. The cytotoxicity was decreased with the increasing PEG content. The particle size of the complex was increased with the increasing PEG at an N/P ratio of 3.0, while the zeta potentials were decreased. The (99m)Tc labeled complexes were transfected into HepG2 and HeLa cells, while the GFP reporter genes were mainly expressed in the HepG2 cells. The in vivo data was achieved in ALB/c-Ha-ras transgenic mice. (99m)Tc labeled GPP(50)/DNA was injected into the mice via the tail vein, and the gamma images were acquired at 5, 15 and 30 min. The (99m)Tc labeled complexes were mainly localized in the heart and liver, and they were excreted through the kidneys. The GFP gene was mainly expressed in the proliferating cells at the tumor periphery. This result was confirmed by PCNA staining. The GPP(50)/DNA complexes were bound to ASGP-R of the proliferating hepatocytes in vitro and in vivo. The present results demonstrate the feasibility of nonviral gene transfer using galactosylated PEI-PEG in vivo. PMID- 16253377 TI - Development of novel sustained-release system, disintegration-controlled matrix tablet (DCMT) with solid dispersion granules of nilvadipine. AB - The goal of this study is to develop a novel sustained-release (SR) system for poorly water-soluble drugs by applying solid dispersion (SD) technique for improving the solubility. The developed SR system, disintegration-controlled matrix tablet (DCMT), consists of hydrogenated soybean oil (HSO) as wax and SD granules containing low-substituted hydroxypropylcellulose (L-HPC) as a disintegrant. In this study, nilvadipine (NiD) was chosen as a model compound. Sustained-release profiles of NiD from DCMT were identically controlled in several dissolution mediums in spite of varying pH and agitation speed. The release of NiD from DCMT was sustained more effectively by increasing the amount of wax or by decreasing the amount of disintegrant, and supersaturation of NiD was achieved without any re-crystallization in dissolution medium. The release rate of NiD from DCMT was controlled by the disintegration rate of tablet. The release profile of NiD was described by the Hixson-Crowell's model better than zero-order kinetics, first-order kinetics and Higuchi's model, which supports that the release of NiD from DCMT is regulated by the disintegration of the tablet. From this study, it was clarified that DCMT was one of the promising SR systems applying SD for the poorly water-soluble drugs. PMID- 16253378 TI - TP10, a delivery vector for decoy oligonucleotides targeting the Myc protein. AB - One approach to investigate gene function, by silencing the activity of certain proteins, is the usage of double stranded decoy oligodeoxynucleotides (ds decoy ODNs). Decoy, in this sense, is ds ODNs bearing the consensus binding sequence for a DNA-binding protein. This can be used in clinical settings to attenuate the effect of overexpressed transcription factors in tumor cells. We here choose to target the oncogenic protein Myc. Since oligonucleotides are poorly internalized to cells, a cell-penetrating peptide, TP10, was coupled to the Myc decoy, using two different strategies. Either TP10 was simply mixed with ds decoy ODNs forming complexes through non-covalent electrostatic interactions, or by having a nona nucleotide overhang in one of the decoy strands, and adding a complementary PNA sequence coupled to an NLS sequence and TP10, which could hybridize to the Myc decoy. By using these strategies, uptake was significantly enhanced, especially with the co-incubation approach. Interestingly, various endocytosis inhibitors had no effect on the uptake pattern, suggesting that uptake of these complexes is not mediated via endocytosis. Finally, a decreased proliferative capacity was observed when treating the neuroblastoma cell line N2a with TP10-PNA conjugate hybridized to Myc decoy compared to naked Myc decoy and untreated cells. A dose dependent decrease in proliferation was also observed in MCF-7 cells, when using both strategies. These results suggest an alternative way to efficiently deliver ds ODNs into cells using the cell-penetrating peptide TP10 and prevent tumor growth by targeting the oncogenic protein Myc. PMID- 16253379 TI - Myocardial dysfunction in mitochondrial diabetes treated with Coenzyme Q10. AB - Maternally-inherited diabetes and deafness (MIDD) has been related to an A to G transition in the mitochondrial tRNA Leu (UUR) gene at the base pair 3243. Although some previous articles have reported that this mutation may be a cause of cardiomyopathy in diabetes, the degree of cardiac involvement and a specific treatment has not been established. Here, we reported a case of a patient with MIDD who developed congestive heart failure and the therapeutic usefulness of Coenzyme Q10 (CoQ10). In our patient, after the introduction of Coenzyme Q10 150 mg/day, there was a gradual improvement on left ventricular function evaluated by echocardiography. The fractional shortening (FS) and ejection fraction (EF) increased from 26 to 34% and from 49 to 64%, respectively. No side effects were noted. Three months after CoQ10 discontinuation, the parameters of systolic function evaluated by echocardiography decreased, suggesting that CoQ10 had a beneficial effect. Identification of diabetes and cardiomyopathy due to mitochondrial gene mutation may have therapeutic implications and Coenzyme Q10 is a possible adjunctive treatment in such patients. PMID- 16253380 TI - Improvement in C-reactive protein and advanced glycosylation end-products in poorly controlled diabetics is independent of glucose control. AB - We studied the efficacy of four different treatment regimens (sulphonylurea and metformin+/-acarbose versus glimepiride and rosiglitazone versus glimepiride and bedtime NPH insulin versus multiple actrapid and NPH insulin injections) in poorly controlled type 2 diabetes subjects on hs-CRP, VCAM-1 and AGE at 4, 8 and 12 weeks of treatment. Multiple insulin injections rapidly improved HbA(1c) by 0.6+/-0.9% (p<0.005), 1.2+/-1.3% (p<0.0005) and 1.3+/-1.4% (p<0.0005) at week 4, at week 8 and week 12, respectively. Subjects who continued their existing combination treatment of sulphonylurea, metformin+/-acarbose also showed a significant reduction in HbA(1c) (p<0.05). Although effective in reducing glycemic parameters, there was no reduction in CRP levels in either treatment group. The treatment regimen consisting of rosiglitazone and glimepiride significantly lowered hs-CRP by -2.6 (3.9) mg/L (p<0.05) at week 12 in spite of no improvement in blood glucose. AGE improved in all groups irrespective of type of treatment, glycaemic control and CRP levels. Our data indicate rapid glycaemic control alone does not necessarily result in improvement in markers of inflammation in type 2 diabetes patients. PMID- 16253382 TI - Failure of hepatitis B vaccination to induce either humoral or cellular immune response after liver transplantation for hepatitis B related cirrhosis. PMID- 16253381 TI - The pruritus of cholestasis. PMID- 16253383 TI - Physician dual practice. AB - Physicians employed in government clinics and hospitals also frequently have private practices. The economic theory of such dual practice is relatively limited and recent. We provide a summary and comparison of five models of dual practice, including one we have developed based on total compensation theory and contracting limitations. We also discuss whether theoretical predictions are consistent with empirical evidence from developed and developing countries. We argue that the social trade-off between the benefits and costs of dual practice hinge on the quality of a country's contracting institutions. The conclusion outlines a proposed research agenda for better understanding this widespread phenomenon in the health sector and in other segments of society. PMID- 16253384 TI - Private medical services in the Italian public hospitals: the case for improving HRM. AB - This study explores how Italian public hospitals can use private medical activities run by their employed physicians as a human resources management (HRM) tool. It is based on field research in two acute-care hospitals and a review of Italian literature and laws. The Italian National Health Service (NHS) allows employed physicians to run private, patient-funded activities ("private beds", surgical operations, hospital outpatient clinics, etc.). Basic regulation is set at the national level, but it can be greatly improved at the hospital level. Private activities, if poorly managed, can damage efficiency, equity, quality of care, and public trust in the NHS. On the other hand, hospitals can also use them as leverage to improve HRM, with special attention to three issues: (1) professional evaluation, development, and training; (2) compensation policies; (3) competition for, and retention of, professionals in short supply. The two case studies presented here show great differences between the two hospitals in terms of regulation and organizational solutions that have been adopted to deal with such activities. However, in both hospitals, private activities do not seem to benefit HRM. Private activities are not systematically considered in compensation policies. Moreover, private revenues are strongly concentrated in a few physicians. Hospitals use very little of the information provided by the private activities to improve knowledge management, career development, or training planning. Finally, hospitals do not use private activities management as a tool for competing in the labor market for health professionals who are in short supply. PMID- 16253386 TI - Molecularly imprinted polymers: a bridge to advanced drug delivery. AB - Specific molecular recognition is a fundamental requirement of living systems on which processes as diverse as neural transmittance, respiration, immune defence, cellular differentiation and nutrition rely. It is therefore not surprising that scientists have invested huge amounts of time and effort into harnessing, and more recently mimicking, biological function. A number of synthetic approaches have been developed and one of the most promising of these is molecular imprinting. Molecular imprinted polymers have routinely been used, as robust and effective synthetic molecular receptors, in a diverse range of technologies. But it is perhaps in the area of drug delivery, in particular 'intelligent drug release' and 'magic bullet' drug targeting, that significant future opportunities lie. PMID- 16253385 TI - Neural steroid hormone receptor gene expression in pregnant rats. AB - Estrogen and progesterone play important roles during pregnancy in stimulating the onset of maternal behavior at parturition. The status of receptor expression of these hormones during pregnancy in neural regions that regulate maternal behavior is unclear. The objective of the present study is to characterize changes in neural gene expression of the estrogen receptors alpha and beta (ERalpha and ERbeta) and the progesterone receptor (PR) during the latter part of pregnancy. Brains from primigravid Sprague-Dawley rats were collected on days 15 and 21 of pregnancy. Micropunches of the olfactory bulb (OB), medial preoptic area (MPOA), bed nucleus of the stria terminalis (BnST), hypothalamus (HYP), medial amygdala (MeA), and the temporal cortex (TCx) were analyzed by real-time RT-PCR (Taqmantrade mark) for levels of gene expression. No changes in either ERalpha or ERbeta mRNA levels were detected in any brain region between days 15 and 21 of pregnancy: however, the MPOA had higher levels of both ERalpha and ERbeta than other brain regions. Progesterone receptor mRNA levels, in contrast, declined significantly in the MPOA, HYP, and TCx, between days 15 and 21 of pregnancy (P < 0.05). In addition, the levels of PR mRNA were significantly higher in the HYP and TCx compared to both the OB and MeA. These data indicate that there is a downregulation of PR prepartum and suggest that this decrease may play a role in the disinhibition of maternal behavior at parturition. PMID- 16253387 TI - ES cell technology: an introduction to genetic manipulation, differentiation and therapeutic cloning. AB - ES cells are extraordinary cells, capable of proliferating in a pluripotent state indefinitely and of differentiating spontaneously into all cell types in vivo and many in vitro. However, the manipulation and modification of ES cells by processes such as directed differentiation and genetic modification have placed ES cells at the forefront of many biological studies and could lead to their application in biopharmaceutical areas such as cellular therapy and drug screening. Here we describe some of the ES cell based technologies that have lead to this realisation of ES cell potential. PMID- 16253388 TI - Door-opening motion can potentially lead to a transient breakdown in negative pressure isolation conditions: the importance of vorticity and buoyancy airflows. AB - A patient with severe chickenpox was admitted to a negative-pressure isolation room. He remained sedated, intubated and mechanically ventilated throughout his admission. He was managed only by nurses immune to chickenpox. A non-immune male nurse occasionally handed equipment through the doorway, without entering the room. Ten days later, he also developed chickenpox. Sequencing of viruses from the patient and nurse showed the same rare genotype, indicating nosocomial transmission. An experimental model demonstrated that, despite negative pressure, opening the door could have resulted in transport of infectious air out of the isolation room, leading to a breakdown in isolation conditions. PMID- 16253389 TI - Possible role of calcitonin gene-related peptide in osmoregulation via the endocrine control of the gill in a teleost, the eel, Anguilla anguilla. AB - Osmoregulation is a major challenge in aquatic animals involving a complex endocrine control. We investigated the potential role of calcitonin gene-related peptide (CGRP, a neuromediator in mammals) in the endocrine control of the gill in a teleost, the eel. Transfer from freshwater to seawater induced an hyperosmolality and a concomitant large increase in plasma CGRP levels. Specific CGRP binding sites were characterized in the gill and their number was up regulated after seawater transfer. This study suggests that the endocrine control of gill function during osmoregulation may represent an ancient role of CGRP in vertebrates. PMID- 16253390 TI - Effects of neuropeptide Y on food intake and brain biogenic amines in the rainbow trout (Oncorhynchus mykiss). AB - Neuropeptide Y (NPY) is one of the most potent stimulants of food intake in mammals, but very little is known about NPY actions in fish. The present study investigated the role of NPY in food intake in the rainbow trout (Oncorhynchus mykiss). Food intake was monitored at different times after intracerebroventricular administration of porcine NPY (4 or 8 microg). Both doses significantly increased food intake at 2 and 3 h, and this effect was dose dependent. However, 50 h after administration of NPY, food intake was significantly lower than in control fish, and cumulative food intake had returned to levels similar to those seen in the control group. The NPY antagonist (D Tyr27,36, D-Thr32)-NPY (10 microg) inhibited food intake 2 h after icv administration, but did not block the orexigenic effect of NPY when administered jointly with 4 microg NPY. To identify the NPY receptor subtypes involved in the effects of NPY on food intake, we studied the effects of the Y1 receptor agonist (Leu31, Pro34)-NPY (4 microg), the Y2 receptor agonist NPY(3-36) (4 microg), and the highly specific Y5 receptor agonist (cPP(1-7), NPY19-23, Ala31, Aib32, Gln34) hPP (4 microg). Short-term (2 h) food intake was moderately stimulated by the Y1 agonist, more strongly stimulated by the Y2 agonist, and unaffected by the Y5 agonist. We found that administration of NPY (8 microg icv) had no effect on aminergic systems in several brain regions 2 and 50 h after NPY administration. These results indicate that NPY stimulates feeding in the rainbow trout, and suggest that this effect is cooperatively mediated by Y2- and Y1-like NPY receptors, not by Y5-like receptors. PMID- 16253391 TI - Inflammatory S100A9 and S100A12 proteins in Alzheimer's disease. AB - Inflammation, insoluble protein deposition and neuronal cell loss are important features of the Alzheimer's disease (AD) brain. S100B is associated with the neuropathological hallmarks of AD where it is thought to play a role in neuritic pathology. S100A8, S100A9 and S100A12 comprise a new group of inflammation associated proteins that are constitutively expressed by neutrophils and inducible in numerous inflammatory cells. We investigated expression of S100B, S100A8, S100A9 and S100A12 in brain samples from sporadic and familial (PS-1) AD cases and controls using immunohistochemistry and Western blot analysis. S100B, S100A9 and S100A12, but not S100A8, were consistently associated with the neuropathological hallmarks of AD. Western blot analysis confirmed significant increases in soluble S100A9 in PS-1 AD compared to controls. S100A9 complexes that were resistant to reduction were also evident in brain extracts. A reactive component of a size consistent with hexameric S100A12 was seen in all cases. This study indicates a potential role for pro-inflammatory S100A9 and S100A12 in pathogenesis caused by inflammation and protein complex formation in AD. PMID- 16253392 TI - DHEA improves symptomatic treatment of moderately and severely impaired MPTP monkeys. AB - The steroid dehydroepiandrosterone (DHEA) is abundant in men and women and decreases rapidly during aging. Parkinson's disease (PD) is the second most common neurodegenerative disorder just behind Alzheimer. l-3,4 Dihydroxyphenylalanine (l-Dopa) therapy remains the most effective treatment but many patients develop motor complications. This study investigated the acute effect of DHEA alone and with l-Dopa in 12 females monkeys lesioned with 1-methyl 4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) to model PD. DHEA administration alone improved the mean parkinsonian score at 1, 5 and 15mg/kg in moderately and severely impaired MPTP monkeys and increased blood DHEA concentrations. DHEA with a low dose of l-Dopa increased the l-Dopa effect in moderately and severely impaired MPTP monkeys. DHEA lengthened duration of the effect of the low dose of l-Dopa by 15-45min. DHEA at 1, 5 and 15mg/kg combined with a high dose of l-Dopa did not increase dyskinesias. DHEA could act by reducing inhibitory GABAergic activity in the striatal output pathways. DHEA could also be metabolized into estradiol in the brain and increase acutely dopamine activity. PMID- 16253393 TI - Imaging and detecting molecular interactions of single transmembrane proteins. AB - Single-molecule atomic force microscopy (AFM) provides novel ways to characterize structure-function relationships of native membrane proteins. High-resolution AFM topographs allow observing substructures of single membrane proteins at sub nanometer resolution as well as their conformational changes, oligomeric state, molecular dynamics and assembly. Complementary to AFM imaging, single-molecule force spectroscopy experiments allow detecting molecular interactions established within and between membrane proteins. The sensitivity of this method makes it possible to detect the interactions that stabilize secondary structures such as transmembrane alpha-helices, polypeptide loops and segments within. Changes in temperature or protein-protein assembly do not change the position of stable structural segments, but influence their stability established by collective molecular interactions. Such changes alter the probability of proteins to choose a certain unfolding pathway. Recent examples have elucidated unfolding and refolding pathways of membrane proteins as well as their energy landscapes. We review current and future potential of these approaches to reveal insights into membrane protein structure, function, and unfolding as we recognize that they could help answering key questions in the molecular basis of certain neuro pathological dysfunctions. PMID- 16253394 TI - Radial distribution function descriptors: an alternative for predicting A2 A adenosine receptors agonists. AB - The Radial Distribution Function approach has been applied to the study of the A2 A adenosine receptors agonist effect of 29 adenosine analogues: N6- arylcarbamoyl, 2-arylalkynyl-N6 -arylcarbamoyl, and N6 -carboxamido derivatives. A model able to describe around 85% of the variance in the experimental activity was developed with the use of the mentioned approach. In contrast, no one of nine different approaches, including the use of Galvez Topological Charges indexes, BCUT, Geometrical, 2D autocorrelations, Topological, Randic Molecular profile, WHIM, 3D-MORSE and GETAWAY descriptors were able to explain more than 78% of the variance in the mentioned property with the same number of variables in the equation. Finally, the model support that the bulkiness and stereoselectivity play an important role in the affinity for this receptor in this kind of compounds. PMID- 16253395 TI - [Bioethics at hospital]. PMID- 16253396 TI - Graduate entrants into nursing - are we meeting their needs? AB - Changes to statutory requirements led to the need to redesign the current shortened programme for graduates (SPG) offered by the University of Dundee. A review of the literature revealed very little published material relating to this type of programme and that which did exist was predominantly American in origin. Prior to developing a new programme a study was carried out. The aim of the study was to explore the attitudes and opinions of non-nurse graduates currently undertaking the accelerated pre-registration programme of study offered by the University of Dundee. DESIGN: Purposive sampling was adopted and all students currently undertaking the programme were invited to participate. Data were collected using three focus group interviews with a minimum of five and a maximum of seven participants. All interviews were tape-recorded. The tapes were transcribed and data analysed using a content analysis approach. FINDINGS: Several important themes and sub-themes emerged from the interviews. These were consistent across all groups. The themes were: DISCUSSION: The data offered a rich insight into the attitudes and opinions of students undertaking the current SPG. The findings supported those of previous studies and have been used to inform curriculum development. PMID- 16253402 TI - Insulin regulates protein synthesis rate in leukocytes from young and elderly healthy humans. AB - Immune response is dependent on synthesis and secretion of various proteins. Hence, in vivo determination of protein synthesis rate in leukocytes may reflect the functional activity of these cells under conditions of immune deficiency such as aging. Because (i) insulin stimulates protein turnover in various tissues and (ii) elderly people often suffer from a decrease in insulin sensitivity, we sought to determine the effect of physiological hyperinsulinemia (0.7mU/kg fat free mass/min) on protein synthesis rate in peripheral blood mononuclear cells (PBMCs) and polymorphonuclear neutrophils (PMNs) from 8 adult (24+/-6yr) and 9 elderly (69+/-4yr) healthy subjects under strict euglycemia (0.9g/l). The body mass index was significantly higher in elderly volunteers compared to the younger ones (P<0.05) whereas insulin sensitivity was lower as shown by either the M value (P<0.05) or the glucose disposal rate (GDR)/insulinemia ratio (P<0.05). The density of the insulin receptor at PBMCs and PMNs surfaces was not altered by either age or insulin infusion. In comparison with the baseline values, insulin highly stimulated the fractional synthesis rate (FSR) of mixed PBMCs (4.12+/ 0.56%/d vs. 6.94+/-0.70%/d, P<0.05) and PMNs (1.05+/-0.28%/d vs. 2.44+/-0.41%/d, P<0.05) proteins. No age effect was observed either during the baseline (PBMCs: 4.31+/-0.30%/d, PMNs: 1.55+/-0.28%/d) period or after insulin infusion (PBMCs: 6.86+/-0.69%/d, PMNs: 2.76+/-0.47%/d). We conclude that insulin is able to stimulate protein synthesis in leukocyte populations independently of the age of the subjects. PMID- 16253403 TI - Effects of total enteral nutrition supplemented with a multi-fibre mix on faecal short-chain fatty acids and microbiota. AB - BACKGROUND AND AIMS: Impaired bowel function is frequent in tube-fed patients, and diarrhoea is associated with decreased faecal short-chain fatty acids (SCFAs) concentrations. The aim of this study was to compare the effects of a multi-fibre enriched formula (15 g/l) and a fibre-free isoenergetic and isonitrogenous formula on faecal SCFAs and microbiota in long-term enteral nutrition (EN) patients. METHODS: Fifteen patients [11M/4F, aged 53 (40-73)] on total EN for 43 (1-310) months for dysphagia received a fibre-free formula for 7 days, followed in a random order by either the multi-fibre-enriched formula for 14 days and then the fibre-free formula for 14 days or vice versa. Stool samples were taken at the end of each period for measurement of SCFAs levels and different groups of bacteria. Results were compared with non-parametric tests. RESULTS: After the multi fibre EN, there was a significant median increase in total faecal SCFAs (+84%), butyrate (+20%) and acetate (+147%) compared with baseline. A significant increase in the total number of bacteria as determined with the molecular method was found after the multi-fibre EN period compared with the fibre-free EN period. There were no concomitant changes in the dominant groups of intestinal bacteria. CONCLUSION: In long-term EN patients, a polymeric enteral formula supplemented with a mixture of six fibres increases faecal SCFAs and total number of bacteria, which may contribute to an improved bowel function. PMID- 16253404 TI - Anti-HBV immune responses in rhesus macaques elicited by electroporation mediated DNA vaccination. AB - Electroporation has been shown to be an effective method to improve the efficiency of gene expression and the immunogenicity of DNA vaccines. In order to optimize the procedure and test for its efficacy in more clinically relevant large animal models, we examined the detailed immune responses in rhesus macaques after vaccination intramuscularly with electroporation using the plasmid encoding for HBV preS(2)-S antigen and an adjuvant plasmid encoding for hIL-2 and hIFN gamma. Several important factors were examined, including the dose response relationships, the effect of various prime and boost regimens, and different combinations of electro-pulse parameters. The immune responses were closely followed for more than a year. The results showed that in rhesus macaques, electroporation can significantly enhance the immunogenicity of the DNA vaccines, resulting in greatly improved antibody responses as well as peptide-stimulated IFN-gamma producing T cell responses. In addition, we also reported the different antibody response behaviors resulted from different electro-pulse parameters. The detailed data would be useful to suggest possible optimization strategies for better DNA vaccine efficacy. PMID- 16253405 TI - Radiocesium contamination behavior and its effect on potassium absorption in tropical or subtropical plants. AB - The accumulation and long-term decline of radiocesium contamination in tropical plant species was studied through measurements of gamma-ray spectra from pomegranate (Punica granatum) and chili pepper (Capsicum fructescens) trees. The plants were originally grown at a (137)Cs contaminated site (where a radiological accident occurred in the city of Goiania, Brazil, in 1987), and transplanted to uncontaminated soil, so that the main source of contamination of the new leaves and fruits would be the fraction of the available radiocesium in the body of the plants. Measurements of (137)Cs and (40)K concentrations along the roots, main trunk, twigs, leaves and fruits before and after the transplant process of both plant species indicated a direct competition between Cs and K ions, suggesting that these elements could have a common accumulation mechanism. Cesium transfer factors from soil to pomegranate, green and red chili pepper fruits were evaluated as 0.4 +/- 0.1, 0.06 +/- 0.01 and 0.05 +/- 0.01, respectively. Biological half-life values due to (137)Cs translocation from the tree reservoir (BHL(T)) were calculated as 0.30 years for pomegranate, 0.12 years and 0.07 years for red and green peppers, respectively. PMID- 16253406 TI - Extensor tendon impingement in a gymnast. AB - Wrist injuries in the gymnast are due to the transformation of the upper extremity into a weight bearing entity. Gymnast wrist pain presents a difficult diagnostic and therapeutic challenge. Here, we present a new case of extensor tendon impingement in an elite gymnast. To our knowledge, there is no similar report in the literature. PMID- 16253407 TI - Distribution and characteristics of organic micropollutants in surface sediments from Bohai Sea. AB - Spatial distribution and compositional characteristics of PAHs, DDTs and PCBs in surface sediments from Bohai Sea were investigated. Proportion of LMW PAHs at Jinzhou Bay was significant, due probably to the petrogenic sources from neighboring oil wells and plants, while HMW PAHs were dominant in the other sea areas, inferred pyrogenic origins mainly from coal or petroleum combustion. The average ERL quotient for the PAH species in Qinhuangdao and Liaodong Bay indicated relatively stronger potential ecological risk. The concentration ratios of DDT to metabolites (DDD + DDE) exceeded 1.0 in the coastal areas of Qinhuangdao, Liaodong Bay and Bohai Bay, demonstrated some recent inputs of DDT nearby, and DDD as the major degradation product. The concentrations of PCBs were generally low, however, the contents of DDTs were greater than the ERL guidelines in the coastal areas of Qinhuangdao, Liaodong Bay and Bohai Bay, and suggested the potential ecological risk. PMID- 16253409 TI - Physiotherapy and the shadow of prostitution: the Society of Trained Masseuses and the massage scandals of 1894. AB - In 1894 the Society of Trained Masseuses (STM) formed in response to massage scandals published by the British Medical Journal (BMJ). The Society's founders acted to legitimise massage, which had become sullied by its association with prostitution. This study analyses the discourses that influenced the founders of the Society and reflects upon the social and political conditions that enabled the STM to emerge and prosper. The founders established a clear practice model for massage which effectively regulated the sensual elements of contact between therapist and patient. Massage practices were regulated through clearly defined curricula, examinations and the surveillance of the Society's members. A biomechanical model of physical rehabilitation was adopted to enable masseuses to view the body as a machine rather than as a sensual being. Medical patronage of the Society was courted enabling the Society to prosper amongst competing organisations. Using Foucault's work on power we explore the contingent nature of these events, seeing the massage scandals in context with broader questions of sexual morality, professionalisation and expertise in the late nineteenth century society. We argue that many of the technologies developed by the founders resonate with physiotherapy practice today and enable us to critically analyse the continued relevance of the profession to contemporary healthcare. PMID- 16253408 TI - Field assessment of oxytetracycline exposure to the freshwater macrophytes Egeria densa Planch. and Ceratophyllum demersum L. AB - In a microcosm study, two aquatic macrophytes, Egeria densa and Ceratophyllum demersum were exposed to nominal concentrations of 0, 5, 20, 50, and 250 microg/L oxytetracycline (n=3), plus 20 microg/L oxytetracycline amended with additional nitrogen (N) and phosphorus (P). Responses were monitored bi-weekly over a six week exposure period. Both plant species exhibited a significant decline in growth in the 250 microg/L oxytetracycline and the N- and P-amended units. Decreased light penetration resulting from accumulating oxytetracycline by products appears to be the primary modifier in the growth of these plants. Increased susceptibility to oxytetracycline exposure was noted in some paired plantings (e.g., E. densa root development), relative to individual plants in these treatments, however, no clear explanation for this response is available. Based on the toxicity data generated in this study, we estimate that current concentrations of oxytetracycline in freshwater environments do not pose a direct risk to E. densa and C. demersum. PMID- 16253411 TI - Clinical variables associated with early administration of antipsychotics in bipolar mania. AB - This study evaluated the clinical variables associated with the early administration of antipsychotics for the treatment of bipolar mania. 63 out of 97 inpatients with bipolar I disorder according to the DSM-IV criteria and who were consecutively admitted to the Kangnam St. Mary's Hospital, between September 2002 and March 2004, were included in the study. Regression analysis was run on the clinical variables in order to identify the factors affecting the early administration of antipsychotics. The route of admission and exposure to antipsychotics at the last follow-up were found to be associated with the early administration of antipsychotics (odds ratios=33.0 and 13.1, respectively). This study suggests that some clinical factors in bipolar mania might be linked to the earlier antipsychotic treatment. However, more studies will be needed to confirm this finding. PMID- 16253410 TI - Nutritional aspects modulating brain development and the responses to stress in early neonatal life. AB - Nutrition is one of the critical factors insuring adequate growth and development in all species. In particular, brain development is sensitive to specific nutrient intake such as proteins and lipids, which are important for cell membrane formation and myelinization. Carbohydrate intake insures adequate short term energy supply, but has important effects on the activity of the hypothalamic pituitary-adrenal (HPA) axis to regulate stress responsiveness. This review focuses on the effects of carbohydrates and fat on the activity of the HPA axis as well as other brain-related functions such as pain modulation, neuropeptide and neurotransmitters release, and some aspects related to cognitive functions. The role of leptin, DHA and AA as mediators of the effects of fat on the brain is discussed. PMID- 16253412 TI - Novel approach on the risk assessment of oxidized fats and oils for perspectives of food safety and quality. I. Oxidized fats and oils induces neurotoxicity relating pica behavior and hypoactivity. AB - Food poisoning caused by deteriorated fat and oil in instant noodles was first reported in Japan approximately 40 years ago. In these cases, many people developed neurotoxic symptoms such as emesis and discomfort. The degree of oxidation of the fat and oil in the instant noodles that induced food poisoning was at least 100 meq/kg in peroxide value (PV). No general toxicity studies with animals, however, have examined the toxicity of fat and oil oxidized to that extent. In this study, pica behavior, a behavior characterized by eating a nonfood material such as kaolin and that relates to the degree of discomfort in animals, and alterations of locomotor activity of rats eating deteriorated fat and oil were measured. The groups fed fat and oil with at least 138.5 meq/kg PV consumed significantly more kaolin compared to the control group. Furthermore, rats that ate deteriorated fat and oil with at least 107.2 meq/kg PV had significantly decreased locomotor activity compared to control rats. These phenomena suggest that oxidized fat and oil with at least 100 meq/kg PV induce neurotoxicity. The toxicity of oxidized fat and oil has only been addressed using general toxicity tests, but the present results reveal the importance of evaluating toxicity by using other measures. PMID- 16253413 TI - Preparing not to move: does no-response priming affect advance movement preparation processes in a response priming task? AB - The response-priming paradigm is frequently employed to study motor control by means of behavioural as well as psychophysiological methods. In this paradigm, response performance is studied under various preparation conditions, but in all cases participants are generally primed for action. This setting neglects an essential ability of the motor system in everyday situations, i.e. the efficient execution of a movement with and without warning. In this context the present experiment assessed the behavioural effects of a modified response-priming paradigm, in which participants were asked to execute movements when primed for action, and when the prime predicted that no response would be required. Thirty volunteers participated the experiment, which tested whether the insertion of no response trials would affect the validity effect, and assessed the differences between invalid trials and reactions performed subsequent to no-response primes. The data showed that the validity effect is not modulated by the added no response condition, which suggests that movement preparation mechanisms can be studied with this particular task. In addition, responses in invalid trials were substantially different from responses executed after no-response primes. These results are interpreted in relation to psychophysiological evidence on advance movement preparation. It is proposed that the performance differences between response- and no-response priming conditions may be due to different preactivations of motor regions evoked by the prime stimuli, a hypothesis to be addressed with psychophysiological measures in the future. PMID- 16253414 TI - Antihypertensive therapy in patients with pre-eclampsia: A prospective randomised multicentre study comparing dihydralazine with urapidil. AB - BACKGROUND: Drug treatment is imperative for pregnant women with pregnancy induced hypertension (PIH) and pre-eclampsia. For more than 40 years, dihydralazine has been the drug of choice for this indication. Another particularly effective and better tolerable antihypertensive is urapidil. Yet only a few studies on limited patient collectives have been published on the clinical experience with urapidil in PIH. METHODS: Urapidil was interindividually compared to dihydralazine in a total of 42 patients, at six participating clinical centres. Patients were randomly assigned to the treatment groups. Urapidil was administered at an initial dose of 12.5-25mg, dihydralazine was administered at a uniform initial dose of 5mg. Patients were closely monitored for the initial 24h of therapy. Until delivery and in the postpartal phase, mother and baby underwent four additional follow-up checks at regular intervals. RESULTS: Either drug was effective in lowering BP. Urapidil treatment proved to be better controllable. There were clear differences as to tolerability. In the urapidil group, one patient complained of headaches. In the dihydralazine group, six patients experienced adverse occurrences. Under dihydralazine treatment, some marked HR increases occurred, interpretable as reflectory tachycardia. CONCLUSIONS: Urapidil proved to be equally effective as dihydralazine in lowering BP in patients with pre-eclampsia, but showed a better controllability and tolerability. Urapidil can hence be recommended as a promising alternative for patients with PIH. PMID- 16253415 TI - A comparison between two prokaryotic potassium channels (KirBac1.1 and KcsA) in a molecular dynamics (MD) simulation study. AB - The two potassium ion channels KirBac1.1 and KcsA are compared in a Molecular Dynamics (MD) simulation study. The location and motion of the potassium ions observed in the simulations are compared to those in the X-ray structures and previous simulations. In our simulations several of the crystallography resolved ion sites in KirBac1.1 are occupied by ions. In addition to this, two in KirBac1.1 unresolved sites where occupied by ions at sites that are in close correspondence to sites found in KcsA. There is every reason to believe that the conserved alignment of the selectivity filter in the potassium ion channel family corresponds to a very similar mechanism for ion transport across the filter. The gate residues, Phe146 in KirBac1.1 and Ala111 in KcsA acted in the simulations as effective barriers which never were passed by ions nor water molecules. PMID- 16253416 TI - Effective factors in thermostability of thermophilic proteins. AB - Thermostability of proteins in general and especially thermophilic proteins has been subject of a wide variety of studies based on theoretical and experimental investigation. Thermostability seems to be a property obtained through many minor structural modifications rather than certain amino acids substitution. In comparison with its mesophile homologue in a thermostable protein, usually a number of amino acids are exchanged. A wide variety of theoretical studies are based on comparative investigation of thermophilic proteins characteristics with their mesophilic counterparts in order to reveal their sequences, structural differences and consequently, to relate these observed differences to the thermostability properties. In this work we have compared a dataset of thermophilic proteins with their mesophilic homologues and furthermore, a mesophilic proteins dataset was also compared with its mesophilic homologue. This strategy enabled us first, to eliminate noise or background differences from signals and moreover, the important factors which were related to the thermostability were recognized too. Our results reveal that thermophilic and mesophilic proteins have both similar polar and nonpolar contribution to the surface area and compactness. On the other hand, salt bridges and main chain hydrogen bonds show an increase in the majority of thermophilic proteins in comparison to their mesophilic homologues. In addition, in thermophilic proteins hydrophobic residues are significantly more frequent, while polar residues are less. These findings indicate that thermostable proteins through evolution adopt several different strategies to withstand high temperature environments. PMID- 16253417 TI - Eco-efficiency in extended supply chains: a case study of furniture production. AB - This paper presents a methodology about how eco-efficiency in extended supply chains (ESCs) can be understood and measured. The extended supply chain includes all processes in the life cycle of a product and the eco-efficiency is measured as the relative environmental and value performance in one ESC compared to other ESCs. The paper is based on a case study of furniture production in Norway. Nine different environmental performance indicators are identified. These are based on suggestions from the World Business Council for Sustainable Development and additional indicators that are shown to have significant impacts in the life cycle of the products. Value performance is measured as inverse life cycle costs. The eco-efficiency for six different chair models is calculated and the relative values are shown graphically in XY-diagrams. This provides information about the relative performance of the products, which is valuable in green procurement processes. The same method is also used for analysing changes in eco-efficiency when possible alterations in the ESC are introduced. Here, it is shown that a small and realistic change of end-of-life treatment significantly changes the eco efficiency of a product. PMID- 16253418 TI - Identifying areas of high herpetofauna diversity that are threatened by planned infrastructure projects in Spain. AB - A major task related to conservation is to predict if planned infrastructure projects are likely to threaten biodiversity. In this study we investigated the potential impact of planned infrastructure in Spain on amphibian and reptile species, two highly vulnerable groups given their limited dispersal and current situation of population decline. We used distribution data of both groups to identify areas of high herpetofauna diversity, and compared the locations of these areas with the locations of the planned road, high-speed train railway and water reservoir network. Four criteria were used for this identification: species richness, rarity, vulnerability, and a combined index of the three criteria. From a total of 1441 cells of 20 x 20 km, areas of high diversity were defined as those cells whose ranked values for the different criteria included either all species or all threatened species. The combined index provided the smallest number of cells needed to retain all threatened species (1.7 and 2.6% of the cells for amphibian and reptile species, respectively). Coincidences between these high diversity areas and cells including planned infrastructures denominated 'alert planning units'-were 35.4% for amphibians and 31.2% for reptiles. Mitigation of the potential impacts would include actions such as barriers to animal access to roads and railways and ecoducts under these constructions. Our approach provides conservation authorities information that can be used to make decisions on habitat protection. A technique that identifies threats to herpetofauna before they occur is also likely to improve the chance of herpetofauna being protected. PMID- 16253419 TI - The conservation benefits of cost-effective land acquisition: a case study in Maryland. AB - Economic theory asserts that to achieve maximum conservation benefits land acquisition needs to be cost effective. Yet the most common planning technique used by land conservation organizations is 'benefit-targeting' that focuses only on acquiring parcels with the highest benefits and ignores costs. Unlike most of the literature which focuses on covering problems, this research applies optimization techniques to achieve maximum aggregate conservation benefits for an ongoing land acquisition effort in the Catoctin Mountain Region in central Maryland. For this case study, optimization yields additional conservation benefits worth an estimated 3.1-3.9 million US dollars or achieves the same level of conservation benefits but at a cost savings ranging from 0.9 to 3.5 million US dollars, depending on the initial budget size. Finally, the highest efficiencies are achieved in low budget scenarios, like those most prevalent in conservation efforts. PMID- 16253420 TI - Differential regulation of corticotropin-releasing factor receptor type 1 (CRF1 receptor) mRNA via protein kinase A and mitogen-activated protein kinase pathways in rat anterior pituitary cells. AB - Corticotropin-releasing factor (CRF) receptor type 1 (CRF(1) receptor) mRNA levels are down-regulated by CRF via the cyclic AMP-protein kinase A (PKA) pathway. In this study, we focused on the involvement of both the mitogen activated protein (MAP) kinase pathway and PKA in this regulation. Real-time PCR (RT-PCR) revealed that a MAP kinase, extracellular signal-regulated kinases 1/2, pathway was also involved in the down-regulation of CRF(1) receptor mRNA levels by CRF in the rat anterior pituitary (AP). Down-regulation of CRF(1) receptor mRNA levels was caused by a post-transcriptional system such as mRNA degradation, as incubation with CRF significantly decreased the half-life of CRF(1) receptor mRNA. Furthermore, pre-treatment with a PKA inhibitor completely blocked CRF induced CRF(1) receptor mRNA destabilization, while pre-treatment with an extracellular signal-regulated kinases 1/2 inhibitor had no inhibitory effect. These results suggested that in the rat AP, down-regulation of CRF(1) receptor mRNA levels is caused by mRNA degradation via PKA, but not by the MAP kinase pathway. PMID- 16253421 TI - Cancer chemopreventive effect of orally administrated lupane-type triterpenoid on ultraviolet light B induced photocarcinogenesis of hairless mouse. AB - Ultraviolet light is the most common cause of skin cancers in human and several effects of ultraviolet light B (UVB) are thought to contribute to skin photocarcinogenesis. The generation of seven triterpenoids from the cones of Liquidamber styraciflua (Hamamelidaceae were examined for their nitric oxide (NO) scavenging activity as the first screening of an in vivo mouse skin anti initiating assay. 3beta,25-epoxy-3alpha-hydroxylup-20(29)-en-28-oic acid (1), which showed the strongest inhibitory effect among these compounds, was studied further for its anti-tumor initiating activity on mouse models initiated with ultraviolet-B (UVB) and promoted with 12-O-tetradecanoylphorbol-13-acetate (TPA). Oral administration of the compound 1 during a period before and after the three times of UVB irradiation exhibited a remarkable effect: administration of 0.0025% solution of the compound 1 given orally only to the test group, which started 1 week before and ended 1 week after the irradiation, showed approximately 50% inhibition of tumor incidence and tumor multiplicity in comparison to the control group. Thus, compound 1 could act as a potent anti-tumor initiator in UVB radiation photocarcinogenesis. PMID- 16253422 TI - Kinetics of acid-catalyzed cleavage of cumene hydroperoxide. AB - The cleavage of cumene hydroperoxide, in the presence of sulfuric acid, to form phenol and acetone has been examined by adiabatic calorimetry. As expected, acid can catalyze cumene hydroperoxide reaction at temperatures below that of thermally-induced decomposition. At elevated acid concentrations, reactivity is also observed at or below room temperature. The exhibited reactivity behavior is complex and is significantly affected by the presence of other species (including the products). Several reaction models have been explored to explain the behavior and these are discussed. PMID- 16253423 TI - Effect of betamethasone on neuropathic pain and cerebral expression of NF-kappaB and cytokines. AB - Glucocorticoids have been used to treat neuropathic pain for many years, but the underlying mechanisms are still unknown. Recent studies indicate that pathological pain states may be mediated by cytokines. We, therefore, examined the effect of betamethasone on neuropathic pain and the relationship between pain behavior and the expression of cytokines in the brain. Rats were given epidural injections of betamethasone (Diprospan) after L5 spinal nerve transection. Mechanical allodynia and thermal hyperalgesia were evaluated on post-operative days 1, 3, 7, 14 and 21 with von Frey and Hargreaves tests. Cerebral expression of NF-kappaB, TNFalpha, IL-1beta and IL-10 was quantified using electrophoretic mobility shift assay (EMSA) or enzyme-linked immunosorbent assay (ELISA). We found that spinal nerve injury caused long-lasting mechanical and thermal hyperalgesia in the hind paw and stimulated the expression of NF-kappaB, TNFalpha, IL-1beta and IL-10 in the brain. A single epidural injection of betamethasone at the time of nerve injury partially inhibited the development of neuropathic hyperalgesia and reduced the subsequent elevated levels of pro inflammatory cytokines in the brain, while stimulating the expression of the anti inflammatory cytokine IL-10. These data support our hypothesis that pro inflammatory cytokines in the brain may mediate the hyperalgesic effects of spinal nerve injury and that the long-acting anti-hyperalgesic effects of epidural glucocorticoid treatment are due to an inhibitory effect on pro inflammatory cytokine levels and a stimulatory effect on anti-inflammatory cytokine levels in the brain. PMID- 16253424 TI - Differential regulation of cyclooxygenase-2 in the rat hippocampus after cerebral ischemia and ischemic tolerance. AB - We investigated the temporal changes and cellular localization of cyclooxygenase 2 (COX-2) in the rat hippocampus during the induction of acquired ischemic tolerance by sublethal ischemia, and compared these changes with those occurring following transient forebrain ischemia. Adult male Sprague Dawley rats were subjected to either 10 min of lethal global ischemia with or without 3 min of sublethal ischemic preconditioning, or 3 min of ischemia only. A short (3 min) cerebral ischemia as well as lethal ischemia with preconditioning substantially and significantly upregulated COX-2 expression in dentate granule cells, as confirmed by immunoblot analysis. This became evident by 4 h, peaked at 1-3 days, and returned to the basal level around 7 days. COX-2 expression was also increased in CA2 and CA3 neurons, although with weaker staining intensity, but in CA1 neurons very weak immunoreactivity was transiently observed. In the ischemic hippocampus, however, in agreement with previous reports, COX-2 expression was induced strongly in vulnerable CA1 and hilar neurons as well as in resistant CA3 and dentate granule cells. These data demonstrated that COX-2 expression is upregulated in neuronal subpopulations destined to survive, i.e., in CA3 and dentate granule cells after ischemia and ischemia-tolerance induction, as well as in ischemia-vulnerable neurons, i.e., in CA1 neurons after lethal ischemia, suggesting that hippocampal neuronal subpopulations have differential sensitivity to COX-2 upregulation. PMID- 16253425 TI - CRF is the 'egg and chicken', whereas ACTH and corticosteroids are only 'chickens' in response to stress. PMID- 16253426 TI - Substance use and multiple victimisation among adolescents in South Africa. AB - The aims of the study were to examine the relationship between multiple victimisation and drug use, and the role of drug use and other intra-personal, peer, parental and environmental factors in predicting multiple victimisation among adolescents in South Africa. A cross-sectional design was employed. The participants comprised 1474 male and female adolescents aged between 12 and 17 years, from Durban and Cape Town. They completed questionnaire measures assessing demographic characteristics; self, peer and parental drug use; self and peer delinquency; parental child-centredness and rules; and community drug availability and exposure to violence on television. A measure of multiple victimisation assessed whether or not the respondents had experienced two or more different types of violence in their lifetime. There was a significant association between frequency of tobacco, alcohol and marijuana use and multiple victimisation. Significant predictors of multiple victimisation in multiple logistic regression analyses were variables within intra-personal, peer, parental and environmental domains. Victimisation prevention programmes in South Africa should be comprehensive and target adolescents' drug use as well as their other psychosocial risk factors. PMID- 16253427 TI - Obesity: A reckoning both for genetic immunity to infection and xenogamy. AB - It is hypothesized that the genesis and spread of obesity is predetermined by genetic immunity to infections and subsequent interethnic genetic admixture. As in genetic immunity to infections, obesity is determined by a constitutional incongruence between relevant acting agents and their molecular targets. Because many hormones share their specific receptors with the acting molecules of infectious agents, mutant formation of a life-saving genetic resistance to infection could also create life-threatening immunity to relevant hormones. This kind of individual molecular constitution could spread in a population as a result of natural selection for genetic immunity to infections that had performed differently in ecologically distinct populations. Extraordinary diversity in the obesity course, manifestations and severity of specific affections and their sizes and focal disposition around the body, is created by the inter-ethnic mating of persons with different grades of hormone-receptor incongruence. PMID- 16253428 TI - The role of nuclear factor kappaB on angiogenesis regulation through monocyte chemotactic protein-1 in myeloma. AB - Multiple myeloma is malignant proliferation of plasma cells and plasmacytoid cells. Vascular endothelial growth factor (VEGF) is known to be one of the most important if not the main regulator of physiologic and pathologic angiogenesis which triggers growth, survival and migration of myeloma cells. It has been shown that circulating mature or bone marrow driven endothelial precursor cells play an important role in neovascularisation. In accordance with these observations, current therapeutic approaches to myeloma include VEGF inhibitors. Since angiogenesis inhibitors are heterogeneous in origin and potency, and their growing list includes many products with a different function it would be of benefit to determine the key molecule produced by transformed plasma cells which stimulates bone marrow environment to produce their homing "milieu" secreting different cytokines such as VEGF, IL-6, and monocyte chemotactic protein-1 (MCP 1). This molecule could be nuclear factor kappa B (NF-kappaB). It has been confirmed that myeloma cells express and produce NF-kappaB. It has been established recently that by blocking NF-kappaB production MCP-1 secretion is reduced up to 60%. If so, this would also reduce production of IL-6 and VEGF, since MCP-1 upregulates VEGF and IL-6 production. This way one could make bone marrow bad environment for myeloma cells to settle, followed with no disease progression. Targeting to NF-kappaB intended to inhibits its activation with receptor antagonist would possibly significantly inhibit lipopolysaccharide induced IL-6, MCP-1 and TNF-alpha. All of them being stimulators for VEGF secretion and indirectly activation of angiogenesis. To conclude, angiogenesis could be induced by myeloma cells themselves through NF-kappaB activation pathway and by inhibiting its activation we might prevent myeloma expansion in bone marrow and progression of the disease by decreased MCP-1 secretion. PMID- 16253429 TI - Treatment of acne could lead to the better control of cellular autoimmune disorders. PMID- 16253430 TI - Antihistamines may be used as an "adjuvant" in treatment of all kinds of acne. PMID- 16253431 TI - Is extensive use of the Valsalva maneuver associated with an increased risk for Alzheimer's disease? PMID- 16253432 TI - Donation of supernumerary oocytes for the derivation of nuclear transfer stem cells, in return for the promise of future therapeutic benefits: ethical and moral obligations of medical professionals to the patient. PMID- 16253433 TI - Potential clinical uses of cetirizine in dermatology. PMID- 16253434 TI - Antimicrobial activity of Centaurea diffusa. AB - The methanol soluble fraction of the aqueous extract obtained from the dry heads of Centaurea diffusa demonstrated antimicrobial activity in twelve evaluated microorganisms. PMID- 16253435 TI - Activities of some Brazilian plants against larvae of the mosquito Aedes aegypti. AB - The insecticidal activities of extracts and oils of seventeen medicinal plants of Brazil have been determined using an Aedes aegypti larvicidal bioassay. Oils from Anacardium occidentalis, Copaifera langsdorffii, Carapa guianensis, Cymbopogon winterianus and Ageratum conyzoides showed high activities with LC50 values of 14.5, 41, 57, 98 and 148 microg/l, respectively. The most active ethanolic extract tested was that from the stem of Annona glabra which presented an LC50 value of 27 microg/l. The potential application of cashew nut oil, an industrial by-product with low commercial value, in the control of the vector of dengue and yellow fever, may be proposed. PMID- 16253436 TI - Molluscicidal evaluation of three common plants from India. AB - Mortality caused by the aqueous extract of latex of Thevetia peruviana, Alstonia scholaris and Euphorbia pulcherrima against two harmful freshwater snails, Lymnaea acuminata and Indoplanorbis exustus, is reported. Latices of all the plants at high doses were also lethal to freshwater fish Channa punctatus, which shares the habitat with these snails, but within 24 h, LC90 of snail L. acuminata did not cause any mortality to fishes in a mixed population of snail and fish. Therefore, these plant extracts may eventually be of great value for the control of harmful aquatic snails and other molluscan pests. PMID- 16253437 TI - Anticonvulsant evaluation of safranal and crocin from Crocus sativus in mice. AB - The anticonvulsant activities of Crocus sativus stigma constituents, safranal and crocin, were evaluated in mice using pentylenetetrazole (PTZ)-induced convulsions in mice. Safranal (0.15 and 0.35 ml/kg, i.p.) reduced the seizure duration, delayed the onset of tonic convulsions and protected mice from death. Crocin (200 mg/kg, i.p.) did not show anticonvulsant activity. PMID- 16253438 TI - Antimicrobial and phytochemical studies on Pedilanthus tithymaloides. AB - The antibacterial and antifungal properties of ethanolic extract of the leaves of Pedilanthus tithymaloides and some of its constituents were investigated by the dilution method. PMID- 16253439 TI - Essential oil composition and antimicrobial activity of Oliveria decumbens. AB - The essential oil of Oliveria decumbens was investigated for its components and antimicrobial activity against six bacteria and two fungal strains. The essential oil was characterized by a high amount of oxygenated monoterpene components of which thymol (47.06%) and carvacrol (23.31%) were the major components. The oil exhibited high antimicrobial activity against all tested Gram+ and Gram- bacteria and fungal strains. PMID- 16253440 TI - A time contraction effect of acute tail-pinch stress on the associative learning of rats. AB - The effect of tail-pinch stress interpolated between the saccharin conditioned stimulus (CS) and the illness-inducing unconditioned stimulus (US) during long trace taste-aversion conditioning was examined in young- and old adult rats with a two-cylinder (saccharin versus water) test. A 2 x 2 x 4 factorial ANOVA was performed on percent-preference-for-saccharin data, with age (young, old), stress condition (stressed, non-stressed), and CS-US interval (22.5-, 45-, 90-, and 180 min) being the factors under consideration. The ANOVA yielded only significant main effects of stress condition and CS-US interval. These findings indicate that stress weakens the CS-US association as evidenced by a higher percent preference for saccharin in the stressed rats than in non-stressed rats at all CS-US intervals. A comparison of the stressed and non-stressed conditioned rats with pseudo-conditioned controls showed that the non-stressed rats formed strong aversions up to the 45-min CS-US interval whereas the stressed rats showed no conditioning beyond the 22.5 min CS-US interval, indicating that stress decreases the effective CS-US interval. Results were interpreted in terms of time contraction and an internal biological countdown timer hypothesized to govern processes involved in associative learning over long delays. PMID- 16253441 TI - Decreased cerebral blood flow of the right anterior cingulate cortex in long-term and short-term abstinent methamphetamine users. AB - BACKGROUND: The aim of the current study was to explore changes of relative regional cerebral blood flow (rCBF) in short-term and long-term abstinent methamphetamine (MA) users. METHODS: Relative rCBF in 40 abstinent MA users and 23 healthy comparison subjects was compared by the technetium-99m-hexamethyl propylene amine oxime ((99m)Tc-HMPAO) single photon emission computed tomography (SPECT). Relative rCBF in areas that were found to differ significantly was also compared in groups of MA users with short-term (<6 months) and long-term (>or=6 months) abstinence. RESULTS: MA users showed decreased relative rCBF in the right anterior cingulate cortex (Brodmann area 32) relative to healthy comparison subjects. Long-term abstinent MA users had significantly greater rCBF than short term abstinent MA users. CONCLUSIONS: We report that abstinent MA users have decreased rCBF in the anterior cingulate cortex with smaller relative decreases in subjects with prolonged abstinence. PMID- 16253442 TI - Molecular genetics of Erwinia amylovora involved in the development of fire blight. AB - The bacterial plant pathogen, Erwinia amylovora, causes the devastating disease known as fire blight in some Rosaceous plants like apple, pear, quince, raspberry and several ornamentals. Knowledge of the factors affecting the development of fire blight has mushroomed in the last quarter century. On the molecular level, genes encoding a Hrp type III secretion system, genes encoding enzymes involved in synthesis of extracellular polysaccharides and genes facilitating the growth of E. amylovora in its host plants have been characterized. The Hrp pathogenicity island, delimited by genes suggesting horizontal gene transfer, is composed of four distinct regions, the hrp/hrc region, the HEE (Hrp effectors and elicitors) region, the HAE (Hrp-associated enzymes) region, and the IT (Island transfer) region. The Hrp pathogenicity island encodes a Hrp type III secretion system (TTSS), which delivers several proteins from bacteria to plant apoplasts or cytoplasm. E. amylovora produces two exopolysaccharides, amylovoran and levan, which cause the characteristic fire blight wilting symptom in host plants. In addition, other genes, and their encoded proteins, have been characterized as virulence factors of E. amylovora that encode enzymes facilitating sorbitol metabolism, proteolytic activity and iron harvesting. This review summarizes our understanding of the genes and gene products of E. amylovora that are involved in the development of the fire blight disease. PMID- 16253443 TI - Domain architectural census of eukaryotic gene products containing O-protein phosphatases. AB - Intricate molecular signalling within cellular environment is manifested through phosphorylation of proteins. Regulation of the phosphorylation state is executed through complex networking among kinases and their biochemical antagonists, the protein phosphatases. Protein dephosphorylation in eukaryotic systems is largely performed through four structurally distinct Ser/Thr and Tyr O-protein phosphatase superfamilies. 555 O-protein phosphatases, belonging to the four distinct families, could be identified using sensitive sequence search techniques across five eukaryotic model organisms (yeast, fly, worm, mouse and humans). These phosphatases could be grouped into 49 subfamilies associated with distinct domain architecture and discrete biochemical function. Only five of the architectures are shared across the five eukaryotic genomes. Interestingly, the number of occurrence of tyrosine phosphatases is correlated to the complexity of the genome. Analysis of domain architectures suggests amenability of the tyrosine phosphatases to occur in complex architectures unlike Ser/Thr phosphatases. Domain duplication and shuffling is shown as the customary mechanism for the evolution of the phosphatases. Several architectures are common between humans and other genomes, which are probably non-linearly inherited in humans or specifically lost in several others. PMID- 16253444 TI - Identification of a novel herpesvirus from a California desert tortoise (Gopherus agassizii). AB - Herpesviruses are significant pathogens of tortoises, causing upper respiratory tract disease and necrotizing stomatitis, with infections often associated with high mortality rates. Herpesvirus infection in a captive California desert tortoise (Gopherus agassizii) was detected by light microscopic observation of intranuclear inclusion bodies in various tissues followed by transmission electron microscopic observation of herpesvirus-like particles, and amplification of herpesvirus nucleic acid sequences using polymerase chain reaction. Using an indirect enzyme linked immunosorbent assay, anti-tortoise herpesvirus antibodies were detected one month after initial onset of clinical signs. This novel herpesvirus is distinct from the previously described tortoise herpesvirus (tortoise herpesvirus-1, THV-1) sharing 83% sequence identity of 60 amino acids of a portion of the DNA polymerase gene and 79% sequence identity across 120 amino acids of a portion of the ribonucleotide reductase gene. Similar to THV-1, this novel herpesvirus, tortoise herpesvirus-2 (THV-2), also clusters with the alphaherpesviruses. PMID- 16253445 TI - Changes in gene expression involved in energy utilization during chicken follicle development. AB - Ovarian follicle development in egg-laying species is characterized by rapid growth in 7 days prior to ovulation when DNA and protein synthesis is markedly increased in the granulosa and theca cells. However, energy and substrate sources to facilitate the extensive DNA and protein synthesis necessary for folliculogenesis have not been identified in avian species. The current study was undertaken to investigate the expression profiles of regulatory genes involved in glucose transport, glycolysis and fatty acid oxidation in the follicle membranes from the small white follicle (SWF) to follicle 1 (F1) stages of follicle development. In our analysis of glucose transporter (GLUT) isoform expression, the level of GLUT1 mRNA increased with follicle development while GLUT2, GLUT3 and GLUT8 mRNA levels were unaffected by follicle development. In contrast, the expression patterns of proteins involved in metabolism down-stream of glucose transport, including hexokinase (HK), pyruvate dehydrogenase E1alpha (PDH E1alpha) and citrate synthase (CS), did not vary with the developmental stage of the follicle, even during rapid follicle growth. Expression of genes related to beta-oxidation of fatty acids (carnitine palmityl CoA transferase I and II, l-3 hydroxyacyl CoA dehydrogenase and long-chain acyl-CoA dehydrogenase), for which expression in the ovarian follicles of mammalian species has not previously been studied, was not changed consistently with the follicle development. These results suggest that both glucose and fatty acids might work as energy sources to ensure rapid follicle development in the chicken ovary, even though glycolysis and beta-oxidation are not modulated by follicle development. PMID- 16253446 TI - Increasing the age of gilts at first boar contact improves the timing and synchrony of the pubertal response but does not affect potential litter size. AB - One hundred and ninety-two Large White/Landrace crossbred gilts were used in this study. The study was conducted in two blocks, with 16 gilts allocated to each of six treatments in each block. The treatments compared the effects on puberty attainment of commencing boar exposure at 161, 182 or 203 days of age, and the effect of first mating gilts at either the pubertal or second oestrus on ovulation rate and early embryo survival. Boar contact took place in a detection mating area (DMA), and consisted of 20 min/day of full contact with a vasectomized boar greater than ten months of age. Gilts were artificially inseminated at the allocated oestrus, with the reproductive tracts collected at 22.8+/-0.4 days after first mating (mean+/-S.E.M.), and the numbers of corpora lutea and viable embryos recorded. The age at which gilts attained puberty increased with the age at which boar exposure commenced. Mean gilt ages at puberty were 179.5+/-1.6, 191.7+/-1.2 and 210.3+/-0.9 days, respectively, when boar contact commenced at 161, 182 and 203 days of age, P<0.01. Mean days-to puberty was significantly shorter when boar contact began at 182 and 203 days of age compared to 161 days of age (10.4+/-1.2 and 8.3+/-0.9 days versus 18.9+/-1.5 days, respectively, P<0.01). Similarly, commencing boar exposure at 182 or 203 days of age as opposed to 161 days of age significantly increased the proportion of gilts attaining puberty within 10 days of start of boar exposure (0.67 and 0.70 versus 0.24, P<0.01). Mean days-to-puberty and the synchrony of puberty attainment were similar when boar contact commenced at 182 and 203 days of age. There was no significant effect of mating oestrus or age of gilts at mating on ovulation rate, embryo number or embryo survival. In conclusion, the current data indicate that the timing and synchrony of puberty attainment is significantly improved when first boar exposure of gilts is delayed until 182 days of age. Further, it is evident that within the age range investigated, delaying first mating until the second oestrus does not significantly increase either ovulation rate or embryo number at day 20 post-mating. PMID- 16253448 TI - PEO and MPEG in high drug load extruded and spheronized beads that are devoid of MCC. AB - A means to produce extruded-spheronized beads, devoid of microcrystalline cellulose (MCC) and with a high drug load (greater than 80%, w/w), is presented. Immediate release bead product with a high yield (greater than 60% of 1mm diameter beads) and low friability (mass loss less than 4.0%) that were spherical to the naked eye (roundness score less than 1.20) were obtained. The formulation consists only of water-soluble components, taking advantage of the properties of soluble polyethylene oxide (PEO) and methoxypolyethylene glycol (MPEG). This approach incorporates minimal processing aids, with wetted PEO providing the apparent plasticity and cohesiveness, and MPEG550 providing the apparent self lubricating characteristics necessary for successful extrusion and subsequent spheronization into beads. The success of this approach has important implications in cases where high drug load beads are desired, but where MCC cannot be used due to chemical incompatibility or where complete release cannot be achieved with MCC-containing beads. PMID- 16253447 TI - Melarsoprol-cyclodextrins inclusion complexes. AB - Melarsoprol, a water-insoluble drug, is mainly used in the treatment of trypanosomiasis and has demonstrated an in vitro activity on myeloid and lymphoid leukemia derived cell lines. It is marketed as a very poorly tolerated non aqueous solution (Arsobal). The aim of our work was to develop melarsoprol cyclodextrin complexes in order to improve the tolerability and the bioavailability of melarsoprol. Phase-solubility analysis showed A(L)-type diagrams with beta-cyclodextrin (betaCD), randomly methylated beta-cyclodextrin (RAMEbetaCD) and hydroxypropyl-beta-cyclodextrin (HPbetaCD), which suggested the formation of 1:1 inclusion complexes. The solubility enhancement factor of melarsoprol (solubility in 250 mM of cyclodextrin/solubility in water) was about 7.2x10(3) with both beta-cyclodextrin derivatives. The 1:1 stoichiometry was confirmed in the aqueous solutions by the UV spectrophotometer using Job's plot method. The apparent stability constants K(1:1), calculated from mole-ratio titration plots, were 57 143+/-4 425M(-1) for RAMEbetaCD and 50 761+/-5 070 M(-1) for HPbetaCD. Data from 1H-NMR and ROESY experiments provided a clear evidence of inclusion complexation of melarsoprol with its dithiaarsane extremity inserted into the wide rim of the cyclodextrin torus. Moreover, RAMEbetaCD had a pronounced effect on the drug hydrolysis and the dissolution rate of melarsoprol. However, the cytotoxic properties of melarsoprol on K562 and U937 human leukemia cell lines was not modified by complexation. PMID- 16253449 TI - Microfabricated drug delivery devices. AB - We review newest developments in the design and fabrication of drug delivery devices based on micropatterned structures. Electronic devices have now reached a stage of dimensions comparable to those of biological macromolecules. This raises exciting possibilities for combining microelectronics and biotechnology to develop new technologies with unprecedented power and versatility. While molecular electronics use the unique self-assembly, switching and dynamic capabilities of molecules to miniaturize electronic devices, nanoscale biosystems use the power of microelectronics to design ultrafast/ultrasmall biocompatible devices, including implants, that can revolutionize the field of bioengineering. Thus, in recent years we have seen an explosion in the field of novel microfabricated and nanofabricated devices for drug delivery. Such devices seek to develop a platform of well controlled functions in the micro- or nano-level. They include nanoparticulate systems, recognitive molecular systems, biosensing devices, and microfabricated and microelectronic devices. PMID- 16253450 TI - Simultaneous effects of tocopheryl polyethylene glycol succinate (TPGS) on local hair growth promotion and systemic absorption of topically applied minoxidil in a mouse model. AB - In this study, topical minoxidil solutions supplemented with TPGS in cosolvent systems of various compositions consisting of water, alcohol, and polyethylene glycol 400 were designed to evaluate the efficacy of promoting hair growth after topical application and the safety in terms of the amount of minoxidil absorbed through the skin into the circulation using C57BL/6J mice as a model. The commercial product of 2% Regaine) was used as the positive control. The role, which sulfotransferase activity plays in hair growth with treatment using minoxidil, was determined as well. The results revealed that the addition of 0.5% TPGS was able to enhance the proliferation of hair, but an increase in the amount of TPGS to 2% led to deterioration in the enhancement of hair growth. At the higher added amount (2.0%) of TPGS, the promotion of hair growth was slightly reduced for both cosolvent formulations F1 (100% water) and F3 (100% PEG 400), whereas it was reduced to a greater extent for the cosolvent formulations F8-F10. In comparison, the influences of cosolvent compositions with TPGS amounts of 0.0 and 2.0% on the promotion of hair growth were similar. On the contrary, variability in the promotion of hair growth by different solvent formulations was minimal when the added amount of TPGS was 0.5%. In general, a relationship between hair growth and sulfotransferase activities after topical application of 2% Regaine and minoxidil formulations containing various amounts of TPGS was not demonstrated. Plasma concentrations of minoxidil with 2% Regaine were found to be greater than those of 2% minoxidil in those cosolvent formulations containing various amounts of TPGS, while showing insignificant differences among those 10 cosolvent formulations with a fixed amount of TPGS. A tendency for the plasma concentration of minoxidil to increase after the topical administration of minoxidil formulations containing the higher amount of TPGS (2%) was noted. PMID- 16253451 TI - Antiulcerogenic activity of Alchornea castaneaefolia: effects on somatostatin, gastrin and prostaglandin. AB - The hydroethanolic extract of the leaves (HEL) and bark (HEB) obtained from Alchornea castaneaefolia (Euphorbiaceae) were investigated for their ability to prevent ulceration of the gastric mucosa in animal models. HEL (500 and 1000 mg/kg) and HEB (1000 mg/kg) significantly reduced the gastric injuries induced by the combination of HCl/ethanol and lowered the severity of gastric damage formation induced by indomethacin/bethanechol in mice. Further investigation showed that HEL also inhibited formation of ulcers in mice submitted to stress and pylorus ligature, but HEL did not modify gastric juice parameters in Shay mice. HEL was also effective in promoting the healing process in chronic gastric ulcer induced by acetic acid in rats. An enriched flavonoidic fraction (EFF at dose of 100mg/kg) obtained from HEL reduced gastric lesions induced by HCl/ethanol and indomethacin/bethanechol in mice. Although EFF did not modify the amount of free mucus production by gastric mucosa, it was able to increase prostaglandin production. When administered to rats submitted to ethanol-induced gastric lesions, EFF increased the somatostatin serum levels, while the gastrin serum levels were proportionally decreased. Phytochemical investigation on HEL and EFF led to the isolation of flavonoids glycosides as the main compounds, thus suggesting that these substances may be involved in the observed antiulcer activity. PMID- 16253452 TI - Comparative anti-hyperglycemic potentials of medicinal plants. AB - Validation of the ethnobotanical use of the leaves of Artemisia vulgaris Linn. (Compositae), Eucalyptus tereticornis Sm. (Myrtaceae), Solanum nigrum Linn. (Solanaceae), and Vitex negundo Linn. (Verbenaceae); stems of Nopalea cochinellifera (Linn.) Salm-Dyck (Cactaceae); roots of Imperata cylindrica Beauv. (Gramineae); dried bark of Syzygium cumini (Linn.) Skeels (Myrtaceae) as anti diabetic agents using the oral glucose tolerance test showed that only the bark of Syzygium cumini and the leaves of Vitex negundo and Eucalyptus tereticornis exhibited anti-hyperglycemic activities when fed simultaneously with glucose. At the same dosages of 5 mg/20 g mouse, Syzygium cumini-treated mice showed a significant decrease in blood glucose levels (BGLs) at 30 min (alpha=0.10) and from 45 min onwards at alpha=0.05. Vitex negundo exhibited greater anti hyperglycemic activity than Eucalyptus tereticornis. Both showed a significant decrease in BGLs at 60 min but at alpha=0.05 for Vitex negundo and at alpha=0.07 for Eucalyptus tereticornis. There was no significant lowering in BGLs for Imperata cylindrica and Solanum nigrum while there was even an increase in BGLs for Nopalea cochinellifera and Artemisia vulgaris. PMID- 16253453 TI - Reconstructing shredded documents through feature matching. AB - We describe a procedure for reconstructing documents that have been shredded by hand, a problem that often arises in forensics. The proposed method first applies a polygonal approximation in order to reduce the complexity of the boundaries and then extracts relevant features of the polygon to carry out the local reconstruction. In this way, the overall complexity can be dramatically reduced because few features are used to perform the matching. The ambiguities resulting from the local reconstruction are resolved and the pieces are merged together as we search for a global solution. The preliminary results reported in this paper, which take into account a limited amount of shredded pieces (10-15) demonstrate that feature-matching-based procedure produces interesting results for the problem of document reconstruction. PMID- 16253454 TI - [Mechanisms of resistance to sexual transmission of HIV-1]. AB - Sexual transmission is the most common pathway for HIV-1; nevertheless some individuals remain seronegative despite repeated high risk sexual exposure. These were grouped in cohorts of "highly exposed but persistently seronegative" individuals, mostly prostitutes and flailing couples. Three lines of defence were observed in these cohorts. The first one is the mucosal barrier, the determining factors of which are the type of epithelium (monolayer or multilayer), epithelial integrity, and the pre-existing microflora. The second one is linked to innate immunity directly related to the genetic and/or immune predispositions of the individual: mutations affecting the CCR5 chemokine receptor, secretion of protective soluble factors, and particular HLA alleles. The third one is acquired immunity via the mechanisms of humoral and/or specific cellular immunity. These studies suggest anti HIV-1 vaccinal strategies aiming at a local immunization combining the different types of responses observed in these individuals. PMID- 16253455 TI - [Cryptococcal neuromeningitis in an immunocompetent patient]. AB - The authors report a case of cryptococcal neuromeningitis (CNM) in a 27-year-old man, non HIV-infected, with a normal CD(4) T-lymphocyte count. He had a clinical history of subacute meningitis. The evolution was fatal. CNM is a rare infection the prognosis of which remains bad, even in immunocompetent patient. PMID- 16253456 TI - [Endocarditis due to Pasteurella sp. Two cases]. AB - Human pasteurellosis is, in general, a locoregional infection due to contact with an animal. Systemic infections are rare and endocarditis is exceptionally described. The authors report two new cases of endocarditis due to Pasteurella spp, they then review 29 other published cases. Pasteurella spp. endocarditis presents as an acute form in 64% of cases and affects the aortic as often as the mitral valves. Contact with an animal is documented in 65% of cases. Pasteurella multocida is the most frequent species in this infection. The total death rate is 40% and can reach 57% of cases in case of immunodepression. The bad prognosis of this infection, justifies an early diagnosis and a rapid and adapted but not yet consensual medicosurgical treatment. PMID- 16253457 TI - [Melioidosis]. AB - Melioidosis is an emerging zoonosis, due to Burkholderia pseudomallei, which is a highly invasive, resistant, and resilient soil bacteria, transmitted by cutaneous or airborne route, and is a potential weapon for bioterrorism. Although the agent has been identified all over the world, the human disease is endemic only in SE Asia and Northern Australia, and gained recent interest after the December 2004 tsunami. Human infection can be a very severe systemic disease (mortality 20 to 80%), with protean expression, but the lung is the most affected organ (50%). Pathophysiology remains unclear. Diabetes mellitus is a major risk factor, and is present in half the Asian patients with melioidosis. Recommended antibiotic regimens are expensive, and in severe disease should be prolonged to 20 weeks to reduce the risk of relapse. Prospects for prevention are limited, and no vaccine is available yet. PMID- 16253458 TI - [Evaluation of the relation between consumption of fluoroquinolones and emergence of resistance among Escherichia coli: contribution of observational and quasi experimental studies]. AB - The emergence of Escherichia coli (E. coli) resistance to fluoroquinolones (FQs) increased and spread gradually worldwide since the early 1990s. The selective pressure of FQs is the main mechanism responsible for the emergence of FQ resistance as shown by in vitro studies. Clinical trials are required to prove the causality between exposure to FQs and emergence of resistance. But this would not be ethical in humans. Non experimental studies must answer several principles to establish causality: association, anteriority, and directional change. We described and compared the contribution of observational and quasi-experimental studies implemented to answer several of these principles. Quasi-experimental studies using interventional models (ARIMA models with transfer function), can answer several of these principles, unlike observational studies. Thus, in addition to assessment of the association, they were able to show that the exposure to FQs precedes the emergence of FQ resistance to E. coli. They were also able to estimate the time necessary for the emergence of resistance and the dose effect, and to show if this association was reversible. PMID- 16253459 TI - [Pleuro-pulmonary paragonimiasis]. AB - Paragonimiasis is a food borne zoonosis due to a trematode belonging to the genus Paragonimus. Although present throughout the world, about 90% of the cases occur in Asia where around 20 million people are infected. The parasitic cycle is complex with two different intermediate hosts. Man is infected by ingesting the raw or undercooked flesh of the second host - a freshwater crab or prawn - or possibly of a paratenic mammal host (wild boar), which contains the infective larval stage metacercariae that reaches the lung which is the main target organ. Epidemiological, pathological, and clinical aspects are reviewed. The main symptoms are protracted cough, and recurrent "benign" hemoptysis. Abnormal pleuro pulmonary imaging features are constant, but protean and non-specific, leading to frequent confusion with tuberculosis. Diagnosis is easily achieved by ova search in the sputum or pleural fluid, or by serology. Evolution is usually considered benign, although not well known. Finally, praziquantel is the effective first choice treatment. Some paradoxical aspects of this disease are underlined such as: underdiagnosis despite a very simple diagnostic procedure, or opposite tendencies according to location, either extinction or re-emergence. PMID- 16253460 TI - [Policies for the use of antibiotic in 99 Southwestern French hospitals in 2002]. AB - OBJECTIVE: Increasing antimicrobial resistance in bacteria is a major health problem and requires the implementation of stringent policies to optimize the use of antibiotics. DESIGN: In 2003 the authors conducted a study in southwestern French hospitals, using a questionnaire to assess the implementation of antibiotic policies according to national guidelines issued by the French government in 2002. RESULTS: The most frequent actions quoted by the 99 respondents were: issuing of a list of available antibiotics, issuing of information regarding antibiotic consumption and bacterial resistance, and control of antibiotics dispensation. Local guidelines were available in 45% of hospitals for curative treatment and in 87% for antibioprophylaxis in surgery. The evaluation of antibiotic use and computer links between clinical settings, pharmacy and microbiology lab were the less widespread measures. The number and type of actions were related to hospital size and activity. CONCLUSIONS: These findings support that policies for an appropriate use of antimicrobials should be reinforced by issuing treatment guidelines and specific tools for dispensation and evaluation. This survey also emphasizes the need for appropriate policies relating to the size and medical activities of healthcare institutions. PMID- 16253461 TI - [Infections associated to severe thrombotic events and antiphospholipid antibodies]. AB - The authors present 2 cases of infections in which the presence of antiphospholipid antibodies (APL), anticardiolipin and anti-beta2-GP1, was associated to the occurrence of significant thrombotic events: 1) a 55-year-old male patient whose serology (indirect immunofluorescence) revealed Coxiella burnetii infection (phase 2 antigens) with IgG at 1,600 and IgM at 50 (significant titer: IgG>or=200 and IgM>or=50); 2) and a 20-year-old male patient with a CMV infection confirmed by serology (IgG: 44 U/ml, significant threshold 6, IgM: 2.1 U/ml, significant threshold 0.9). PMID- 16253462 TI - Experimental design for in vitro skin penetration study of liposomal superoxide dismutase. AB - A computer-based technique using a 2((5-2)) fractional factorial design was applied for screening the factors affecting the penetration effectiveness of liposomal recombinant human-Cu/Zn-superoxide dismutase. Unilamellar liposomes, containing recombinant rh-Cu/Zn-SOD in the aqueous core, are aimed at enhancing the penetration of the drug applied topically. Factors that mainly influence the chemical and physical characteristics of liposomes such as charge, molar content of cholesterol, size, surfactant and lipid were evaluated at two levels. In vitro skin penetration studies with pigskin were carried out in Franz-type diffusion cells over a period of 4 and 8h. The response variables, namely the amounts of rh Cu/Zn-SOD penetrated into the different skin layers, were analyzed by ELISA (enzyme linked immunosorbent assay). Analysis of variance showed that the size and the cholesterol content of liposomes as well as the duration of the penetration studies have a statistically significant influence on the amount of protein found in deeper skin layers. PMID- 16253463 TI - Quantitative determination of captopril and prednisolone in tablets by FT-Raman spectroscopy. AB - A procedure for the quantitative determination of captopril and prednisolone in commercial tablets based on partial least squares (PLS) and principal component regression (PCR) treatment of FT-Raman spectroscopic data is described. In the studied medicines active pharmaceutical ingredients (APIs) constitute 4.2-16.7% of the tablet mass. Results obtained from calibration models built using unnormalised spectra were compared with the values found when an internal standard was added to each sample and the spectra were normalised by its selected band intensity at maximum or integrated. To appraise the quality of the models the relative standard error of predictions (RSEPs) were calculated for calibration and testing data sets. For captopril determination these were 1.8 2.2% (2.1-2.3%) and 2.7-3.1% (2.7-3.6%), respectively for the different PLS (PCR) models. For prednisolone these errors amounted to 1.8-2.1% (2.6-3.5%) and 3.2 3.7% (3.7-5.9%), respectively. Three commercial preparations of captopril containing 12.5mg and one 25mg of API per tablet were quantified using developed models. Found captopril contents, calculated versus results of iodometric titration, was equal 99.2-101.2% (99.2-102.0%), for the different PLS (PCR) calibration models and the different preparations. Quantification of prednisolone tablets, declared content 5mg per tablet, on the basis of PLS (PCR) models gave API amount, calculated versus results of UV-vis method, in the 99.0-101.0% (98.0 102.0%) range. PMID- 16253464 TI - The application of LC-NMR and LC-MS for the separation and rapid structure elucidation of an unknown impurity in 5-aminosalicylic acid. AB - This work has demonstrated the usefulness of combining liquid chromatography nuclear magnetic resonance spectroscopy (LC-NMR) and liquid chromatography-mass spectrometry (LC-MS) methodologies for a rapid identification of an unknown impurity (N1) in the drug 5-aminosalycilic acid. Complementary information obtained from the two methods has revealed plenty of structural information and led to the fast on-line structure determination of N1 prior to its isolation and purification. The analysis of LC-NMR and LC-MS spectra revealed that N1 and 5 aminosalycilic acid are structurally closely related compounds. The structure of N1 was later confirmed by high-resolution NMR spectroscopy of the isolated compound and the atom assignment was made. The approach described here has potential for 5-aminosalycilic acid impurity profiling and monitoring the production process. PMID- 16253465 TI - Determination of paeonol and paeoniflorin in Chinese medicine Cortex Moutan and 'Shuangdan' granule by micellar electrokinetic capillary chromatography. AB - An easy, simple and rapid micellar electrokinetic capillary chromatography (MEKC) method was developed for the separation of two active components paeonol (PN) and paeoniflorin (PF) within 7 min. Capillary electrophoresis (CE) was performed using a 50.0 cm (42.0 cm to the detector window) x 75 microm i.d. fused-silica capillary. The optimal running buffer containing 10mM borate and 25 mM SDS at pH 9.54 was employed. The applied voltage 15 kV and the temperature 25 degrees C was used in CE separation. The linearities between peak areas and the concentrations of the analytes were investigated, and they exhibited excellent linear behavior over the investigated concentration ranges (R(2): 0.9945 for PN and 0.9992 for PF). The method was successfully applied to the determination of these two components contained in Cortex Moutan and 'Shuangdan' granule. The average recoveries ranged between 97.6 and 105.3% for PN and 95.3 and 106.1% for PF, respectively. PMID- 16253466 TI - Patient satisfaction at and after discharge. Effect of a time lag. AB - OBJECTIVE: Patient satisfaction is an important outcome measure for evaluating the quality of medical care. It is remarkable that consistently high satisfaction ratings have been reported over the last 30 years. There are indications that the time point of administration of a patient satisfaction questionnaire has an influence on satisfaction ratings. This study aimed at investigating whether the assessment of patient satisfaction at different time points resulted in different outcomes. METHODS: Patient satisfaction was measured twice. The sample consisted of 152 orthopedic patients who filled in the questionnaire at hospital discharge and one to 12 months after discharge. RESULTS: At follow-up, satisfaction ratings decreased significantly. Satisfaction with postoperative information decreased the most after discharge. CONCLUSION: The results of this study indicate that the time point of administration of a patient satisfaction questionnaire does influence satisfaction ratings. PRACTICE IMPLICATIONS: Patient satisfaction outcomes collected during hospitalization and after discharge may not be interpreted similarly. PMID- 16253467 TI - A new educational program for patients suffering from sleep apnea syndrome. AB - OBJECTIVE: Patients' knowledge and beliefs about their chronic illnesses such as OSAS are known to influence a range of health related variables, including treatment compliance. It is for that reason that this interdisciplinary program centred on the therapeutic education of the patients was conceived by two medical teams with the aim of creating a new coverage of the patients affected of OSAS. METHODS: Thirty-five OSAS patients were consecutively collected among those regularly followed at the Service of pneumology outpatient facility. The psycho educational methodology as well as the courses are described in detail. Patients were admitted for 36 h at the Service of Therapeutic Education for Chronic Diseases for both individual and group multidisciplinary approach to their disease. Groups of 3-4 patients discuss benefits, inconvenience as well as CPAP representations and manipulation of the device in small workshops. RESULTS: The therapeutic objectives, the educational methods, the program evaluation are presented. Preliminary results show a clinical improvement of the duration of CPAP use (0.7 +/- 0.2 h/night). 24% of patients increased the CPAP use more than one hour per night 3 months later. The Epworth sleepiness scale is significantly improved 3 months later (p < 0.05). CONCLUSION: We describe a new therapeutic educational interdisciplinary program for patients affected by OSAS. Its efficacy will be tested prospectively in a larger group and in the long term. PRACTICE IMPLICATIONS: The educational methods were applicable to the patients affected of OSAS bringing new specific strategies to improve the CPAP use. Patients had a positive attitude towards the CPAP treatment. PMID- 16253468 TI - The impact of depressive symptoms and psychosocial factors on medication adherence in cardiovascular disease. AB - OBJECTIVE: This study sought to determine the influence of depression and psychosocial factors on medication adherence in cardiovascular disease. METHODS: A questionnaire including measures of depression, beliefs about medicines, health locus of control and adherence to medication (self-report) was completed by 122 outpatients attending a cardiac clinic. RESULTS: Analysis revealed that 14.8% of participants were non-adherent with their cardiovascular medication and 41.7% had scores indicative of depressive symptoms as determined by the Center for Epidemiological Studies Depression Scale (CES-D). Higher scores on this scale and strong concern scores on the Beliefs about Medicines Questionnaire about the potential adverse effects of using medication as prescribed were found to be associated with self-reported non-adherence. DISCUSSION AND CONCLUSION: These findings imply that the relationship between depressive symptoms in cardiovascular patients, together with certain psychosocial factors, could have negative consequences for adherence to medication. PRACTICE IMPLICATIONS: Given that there is emerging evidence to suggest an association between depression and medication non-adherence, healthcare professionals should consider this when dealing with cardiovascular patients. PMID- 16253469 TI - [Regional block for the elbow, wrist and hand]. PMID- 16253470 TI - [Recommendations for the good use of the pneumatic tourniquet in surgery]. PMID- 16253471 TI - [Axillary block]. PMID- 16253472 TI - [Locoregional intravenous anesthesia]. PMID- 16253473 TI - [Systemic toxicity with mepivacaine following axillary block in a patient with terminal kidney failure]. AB - As onset time and duration of sensory block are intermediate, mepivacaine is widely used for regional anaesthesia. Few reports of systemic adverse effects are available following nerve blockade with mepivacaine. We report the case of a 54 year-old patient suffering from terminal renal failure who needs the confection of an arteriovenous shunt under axillary brachial plexus block. At completion of the injection of 25 ml (375 mg) of 1.5% mepivacaine the patient presented dysarthria, mental confusion followed by a loss of verbal contact and agitation, but no convulsion or cardiac dysrythmia. The patient received midazolam and surgery was planned the following day under general anaesthesia. Plasma mepivacaine concentration at time of neurological signs was measured at 5.1 microg/ml. Prevention and treatment of systemic toxic effects after regional anaesthesia are discussed. PMID- 16253474 TI - [Peripheral nerve block: yesterday's facts and tomorrow's challenges]. PMID- 16253475 TI - [Block of the branches of the brachial plexus and brachial canal]. PMID- 16253476 TI - Genetic susceptibility to infectious diseases. AB - The clinical outcome of infectious disease (ID) is determined by a complex interaction between microorganism, host genetic factors and environment. Epidemiological studies have revealed differences within and between populations exposed to the same infectious agent, and in the prevalence or severity of the disease, underlying the relevance of the genetic background. Population genetic studies have estimated the genetic component (susceptibility) in the ID determination and have identified some susceptibility gene(s)/polymorphism(s). This paper describes the methods used in genetic epidemiology. Complex segregation analysis is used to define genetic models. Parametric linkage analysis and association studies are used to identify polymorphisms strongly linked to the disease. Genome-wide scan and microarray technology are used to map and identify major genes for ID. Future developments will identify subgroups of subjects at different risks of developing ID. PMID- 16253477 TI - Interference with CD4+CD25+ T-cell-mediated tolerance to experimental autoimmune thyroiditis by glucocorticoid-induced tumor necrosis factor receptor monoclonal antibody. AB - Experimental autoimmune thyroiditis (EAT), a murine model for Hashimoto's thyroiditis, is inducible with mouse thyroglobulin (mTg), and characterized by mononuclear cell infiltration and destruction of the thyroid gland. Pretreatment with mTg leads to CD4+CD25+ T-cell-mediated resistance to subsequent EAT induction. We have recently demonstrated that in vivo administration of a monoclonal antibody (mAb) to CD137, a member of the tumor necrosis factor receptor (TNFR) family, interferes with both the development and mediation of induced EAT tolerance. Here, we examined the influence of another TNFR family member, glucocorticoid-induced TNFR (GITR), which has been reported to modulate the function of CD4+CD25+ T cells in other models. We found that in vivo administration of GITR mAb inhibited EAT tolerance induction and abrogated established tolerance, enabling thyroiditis induction. In in vitro assays, GITR mAb inhibited suppression of mTg-primed T cells by CD4+CD25+ T cells isolated from mTg-pretreated mice. The target of GITR mAb appears to be CD4+CD25- T cells, rather than CD4+CD25+ T cells from tolerized mice, suggesting that GITR signaling likely interferes with EAT tolerance by enabling thyroiditogenic T cells to circumvent suppression by CD4+CD25+ regulatory T cells. PMID- 16253478 TI - Tardive Dyskinesia: analysis of outpatients with schizophrenia from Africa and the Middle East, Asia, Central and Eastern Europe, and Latin America. AB - The point prevalence of Tardive Dyskinesia (TD) in schizophrenia outpatients (n=6981) participating in a study of health outcomes was 8.9%. Duration of diagnosis, age, and prior use of typical antipsychotics were diagnostic indicators of TD in this population, with male sex further increasing risk. This study provides new data in a non-Western population with a unique regional geography comparison. PMID- 16253479 TI - Psychopathy and institutional outcome in patients with schizophrenia in forensic settings in the UK. AB - This study examined the institutional outcomes (12 weeks post-admission) of 134 male patients with DSM-IV schizophrenia assessed using the Psychopathy Checklist: Screening Version (PCL: SV) in a medium secure unit in the UK. High psychopathy scorers (HP) were more likely to be violent, non-compliant with programmes, engage in substance misuse violations, have criminal attitudes/peers and have low levels of insight into risk and violence. Psychopathy was also a modest predictor of institutional outcome in these domains. PMID- 16253480 TI - Clinical guidelines for the management of cancer patients with neutropenia and unexplained fever. PMID- 16253483 TI - Mapping of temporal and parietal cortex in progressive nonfluent aphasia and Alzheimer's disease using chemical shift imaging, voxel-based morphometry and positron emission tomography. AB - Little and controversial evidence is available from neuroimaging studies in progressive nonfluent aphasia (PNA). The goal of this study was to combine information from different imaging modalities in PNA compared with Alzheimer's disease (AD). Chemical shift imaging (CSI), voxel-based morphometry (VBM) and fluorodeoxyglucose positron emission tomography (FDG-PET) were used in 5 PNA, 10 AD patients and 10 normal subjects. Group comparisons revealed left anterior lateral temporal abnormalities (BA20/21) in PNA using CSI, VBM and PET in comparison to normal subjects. AD patients showed more limited hypometabolism within the same area. In addition left lateral parietal (BA40) abnormalities were demonstrated in our PNA as well as our AD group using PET and VBM (AD group only). Combining information from all imaging modalities on a single case basis revealed pathology within the left anterior lateral temporal and lateral parietal lobe both in PNA and AD. PNA and AD patients differed significantly, however, with respect to the frequency of medial temporal lobe and posterior cingulate/precuneus involvement. Although our results might not be generalizable to all subgroups of PNA, we conclude that medial temporal and posterior cingulate/precuneus cortex pathology as assessed by CSI and VBM or PET distinguish PNA from AD, whereas lateral temporal and parietal areas are involved in both conditions. PMID- 16253482 TI - Quantitative MRI measures of orbitofrontal cortex in patients with chronic schizophrenia or schizoaffective disorder. AB - The relationship between orbitofrontal cortex (OFC) volumes and functional domains in treatment-resistant patients with schizophrenia or schizoaffective disorder is poorly understood. OFC dysfunction is implicated in several of the behaviors that are abnormal in schizophrenia. However, little is known about the relationship between these behaviors and OFC volumes. Forty-nine patients received magnetic resonance imaging scanning as part of a double-blind treatment study in which psychiatric symptomatology, neuropsychological function, and aggression were measured. OFC volumes were manually traced on anatomical images. Psychiatric symptomatology was measured with the Positive and Negative Syndrome Scale (PANSS). Aggression was measured with the Overt Aggression Scale (OAS) and with the PANSS. Neuropsychological function was assessed using a comprehensive test battery. Larger right OFC volumes were associated with poorer neuropsychological function. Larger left OFC gray matter volumes and larger OFC white matter volumes bilaterally were associated with greater levels of aggression. These findings are discussed in the context of potential iatrogenic effects. PMID- 16253485 TI - Pre-attentive and attentive processing of temporal and frequency characteristics within long sounds. AB - Attention effects on the processing of deviations in the duration and the frequency dimension of a long sound were investigated in three conditions: (1) when auditory stimuli were ignored, (2) when they were attended and frequency dimension was task-relevant, and (3) when they were attended and duration dimension was task-relevant. The mismatch negativity (MMN) of the event-related potential (ERP) to infrequent shortenings of a sound (600 ms vs. 1000 ms) and to infrequent frequency modulations at one of nine possible intervals within the sound (change from 440 Hz to 480 Hz and back to 440 Hz, e.g. in the 600-650 ms interval) was measured. Duration MMN was slightly enhanced when directing attention towards the frequency dimension but notably enhanced when attention was focused on duration. The early phase of frequency-modulated MMN was of equal amplitude in all three conditions, and the late phase was equally enlarged in both attend conditions. Interestingly, MMN to frequency-modulated deviants decreased the later the deviation occurred within the sound; there was no indication for an MMN being present in Ignore condition when frequency modulations occurred 400 ms after sound onset or later. Thus, with increasing temporal distance between the onset of a sound and the onset of a deviation within the sound (e.g. frequency modulation or sound offset), MMN for frequency modulations and duration shortenings decreases. This suggests that the initial part of a sound ( approximately 300 ms) contributes more to the unitary sound representation underlying MMN than the later parts. PMID- 16253484 TI - Stress, strain, and NMR. AB - Experimental and ab initio results that demonstrate the effect of stress on the nuclear magnetic resonance spectra of materials are shown. The design of a cell that generates uniaxial compressive stress is presented, and results on gallium phosphide and lead nitrate single crystals that illustrate the observable results of the stress are shown. Tensors that relate stress and strain to changes in the chemical shielding tensors and the electric field gradient tensors are defined formally. The elements of these tensors are then computed by a density functional theory approach that makes use of planewaves and pseudopotentials. The experimental results are interpreted with the aid of the calculations. Extensions to spinning samples and to the interpretation of optical phenomena in materials are discussed. PMID- 16253486 TI - The difference between partitioning and distribution from a thermodynamic point of view: NSAIDs as an example. AB - Solubility and solvation of some NSAIDs were studied in their non-ionic (aqueous buffers of pH 2.0) and ionic molecular form (pH 7.4) over a wide temperature interval. Absolute scale values for the thermodynamic terms (Gibbs energy, enthalpy and entropy) were obtained. Thermodynamic parameters of the transfer of the molecules from one buffer to the other (representing protonation/deprotonation) were derived. It has been found that the thermodynamic characteristics of solvation (hydration) of (+)- and (+/-)-IBP in the buffers show a difference, which is larger than the experimental error. This may be explained by differences in the association states of the molecules in solution. For the other NSAIDs studied, a correlation between the Gibbs energy of transfer, deltaG(tr) (pH 7.4-->pH 2.0) and the pK(a)-value, and a compensation effect between the enthalpic and entropic terms have been revealed. Thermodynamic aspects of the transfer process from the buffers to n-octanol were analysed. The two types of the transfer processes (non-dissociated molecule to octanol (partitioning), and dissociated form to octanol (distribution)) have essentially different driving forces: partitioning is enthalpy driven, whereas the transfer of the ionic form is entropy driven. The following points are discussed: (a) significance of using water-octanol systems (logP as a measure of drug lipophilicity) to describe biological membranes (lipid systems); (b) differences in thermodynamic aspects of the partitioning/distribution processes of these systems; (c) advantages of the present transfer method approach in comparison with temperature dependencies of logP to analyse the driving forces of partitioning/distribution. PMID- 16253487 TI - Biodegradable thermo-sensitive nanoparticles from poly(L-lactic acid)/poly(ethylene glycol) alternating multi-block copolymer for potential anti cancer drug carrier. AB - In order to produce biodegradable thermo-sensitive nanoparticles, alternating multi-block copolymers (MBC) were synthesized by coupling dicarboxylated poly(ethylene glycol) (PEG; M(w) 2000) with poly(L-lactic acid) (PLLA)/PEG/PLLA triblock copolymers. Three different multi-block copolymers were synthesized by varying PLLA molecular weight (800 (MBC1), 1600 (MBC2), and 2800 (MBC3)). The MBC formed self-assembled nanoparticles with a unimodal size distribution during a dialysis process. The nanoparticles (NP) had a spherical shape with a size range of 90-330 nm in diameter and critical aggregation concentrations in a range of 5.6-12.6 microg/mL, depending on PLLA length in MBC. The thermo-sensitivity of MBC NP was monitored by the changes in particle size and interior structure as a function of temperature. The particle size slightly decreased as increasing temperature from 37 to 42 degrees C. The interior structure of the NP responded to temperature by altering microviscosity. The microviscosity, measured by the anisotropy (r value) of a fluorescence probe, of MBC1 NP significantly changed with increasing temperature (r = 0.187 at 25 degrees C and 0.216 at 42 degrees C), while MBC2 and MBC3 showed negligible changes in the microviscosity. This indicates that the temperature-dependent interior structure of the NP relied on the portion of PLLA in MBC. The thermo-sensitivity affected to the drug release behavior and cell cytotoxicity. At 42 degrees C, doxorubicin (DOX) loaded MBC1 NP showed enhanced cytotoxicity (approximately 20 times) against Lewis Lung Carcinoma (LLC) cells when compared to that at 37 degrees C. PMID- 16253488 TI - Improved evaluation of back muscle SEMG characteristics by modelling. AB - Surface EMG (SEMG) as non-invasive method is a valuable tool in functional studies of movement co-ordination. The interpolation of the SEMG power (EMG mapping) gives information about intra- and inter-muscular co-ordination. It has been shown that SEMG maps of low back pain patients and healthy subjects differ. The only major drawback to SEMG is that volume conduction of muscle tissue, fat, and skin decreases the spatial and temporal resolution of signals. To improve the interpretation of SEMG signals, we have applied high pass filtering of cross covariance functions, which has proved to be useful in increasing the spatial resolution, to SEMG data of the back region. Experimental data demonstrate that SEMG signals from the back extensors show only rarely signs of action potential propagation. This behaviour, also described in the literature, can be explained by a model assuming short, deep muscle fibres, having bipolar end effects, with overlapping positions parallel to the fibre direction. This condition is fulfilled by the mm. multifidii et rotatores which are part of the m. erector spinae. Although the model is simplistic, the agreement between simulations and experiments is good. PMID- 16253489 TI - [Is screening for Chlamydia Trachomatis infection in all sexually active adolescent females appropriate?]. PMID- 16253490 TI - FGF upregulates osteopontin in epiphyseal growth plate chondrocytes: implications for endochondral ossification. AB - Fibroblast growth factor receptor 3 (FGFR3) signaling pathways are essential for normal longitudinal bone growth. Mutations in this receptor lead to various human growth disorders, including Achondroplasia, disproportionately short-limbed dwarfism, characterized by narrowing of the hypertrophic region of the epiphyseal growth plates. Here we find that FGF9, a preferred ligand for FGFR3 rapidly induces the upregulation and secretion of the matrix resident phosphoprotein, osteopontin (OPN) in cultured chicken chondrocytes. This effect was observed as early as two hours post stimulation and at FGF9 concentrations as low as 1.25 ng/ml at both mRNA and protein levels. OPN expression is known to be associated with chondrocyte and osteoblast differentiation and osteoclast activation. Unexpectedly, FGF9 induced OPN was accompanied by inhibition of differentiation and increased proliferation of the treated chondrocytes. Moreover, FGF9 stimulated OPN expression irrespective of the differentiation stage of the cells or culture conditions. In situ hybridization analysis of epiphyseal growth plates from chicken or mice homozygous for the Achondroplasia, G369C/mFGFR3 mutation demonstrated co-localization of OPN expression and osteoclast activity, as evidenced by tartarate resistant acid phosphatase positive cells in the osteochondral junction. We propose that FGF signaling directly activates OPN expression independent of chondrocytes differentiation. This may enhance the recruitment and activation of osteoclasts, and increase in cartilage resorption and remodeling in the chondro-osseus border. PMID- 16253491 TI - Changes in expression of extracellular matrix genes, fibrogenic factors, and actin cytoskeletal organization in retinol treated and untreated vocal fold stellate cells. AB - The regulation of extracellular matrix (ECM) constituency is critical in maintaining vocal cord biomechanical viscoelasticity required for phonation. Recently our laboratory successfully isolated and cultured a novel cell called a vocal fold stellate cell (VFSC), thought to play a central role in laryngeal ECM metabolism, aging, scarring and cancer. Our laboratory has shown that these cells undergo transdifferentiation that is partially reversed by exposure to all-trans retinol (ATROH). Here we make the first report on the expression of various ECM components, MMPs, TIMPs, pro-fibrogenic cytokines, and other ECM modulators in transdifferentiated and deactivated VFSCs. We show that VFSCs maintain an ECM expression pattern similar to laryngeal cancer and scars but distinct from tracheal fibroblasts. Exposure to ATROH differentially affects the VFSC expression of ECM components, matrix-regulating enzymes, and fibrogenic factors suggesting that the inhibitory effects of this synthetic cofactor should be studied further in laryngeal fibrosis and scarring. We also show that increased exposure to retinol induces sequential reorganization of the actin cytoskeleton in activated VFSCs. Our findings demonstrate that VFSCs are capable of regulating vocal fold ECM constituency important throughout normal laryngeal development. Furthermore, our results implicate VFSC activation in ECM misregulation which is a hallmark of several laryngeal pathologies. PMID- 16253492 TI - Dense tissue-like collagen matrices formed in cell-free conditions. AB - A new protocol was developed to produce dense organized collagen matrices hierarchically ordered on a large scale. It consists of a two stage process: (1) the organization of a collagen solution and (2) the stabilization of the organizations by a sol-gel transition that leads to the formation of collagen fibrils. This new protocol relies on the continuous injection of an acid-soluble collagen solution into glass microchambers. It leads to extended concentration gradients of collagen, ranging from 5 to 1000 mg/ml. The self-organization of collagen solutions into a wide array of spatial organizations was investigated. The final matrices obtained by this procedure varied in concentration, structure and density. Changes in the liquid state of the samples were followed by polarized light microscopy, and the final stabilized gel states obtained after fibrillogenesis were analyzed by both light and electron microscopy. Typical organizations extended homogeneously by up to three centimetres in one direction and several hundreds of micrometers in other directions. Fibrillogenesis of collagen solutions of high and low concentrations led to fibrils spatially arranged as has been described in bone and derm, respectively. Moreover, a relationship was revealed between the collagen concentration and the aggregation of and rotational angles between lateral fibrils. These results constitute a strong base from which to further develop highly enriched collagen matrices that could lead to substitutes that mimic connective tissues. The matrices thus obtained may also be good candidates for the study of the three-dimensional migration of cells. PMID- 16253493 TI - Predictors of symptom recurrence after low-dose inhaled corticosteroid cessation in mild persistent asthma. AB - In order to identify predictors of recurrence of asthma symptoms after withdrawal of therapy in mild persistent asthmatics, asymptomatic on low-dose inhaled corticosteroids (ICS), we studied 87 asthmatic patients regularly treated with ICS for at least 6 months. At the enrollment visit (T1), 71 on ICS were asymptomatic over the past 3 months and discontinued asthma treatment. Symptoms and PEF were then monitored for up to 3 months or until symptoms recurred (T2). At T1 and T2, all subjects underwent methacholine challenge and sputum induction. Thirty nine out of 71 patients experienced symptom recurrence. At T1, clinical and functional data and sputum eosinophilia between patients with or without recurrence of symptoms were similar. Age > 40 yr, and disease duration > 5 yr were significantly associated with recurrence of asthma symptoms, while the presence of allergic rhinitis, low baseline FEV(1) and untreated time span > 60 months showed a trend to be associated with symptoms recurrence. At T2, symptoms, pulmonary function, bronchial hyperresponsiveness and sputum eosinophilia deteriorated in patients with symptom recurrence but not in patients without symptom recurrence. In conclusion, age and asthma duration were the best predictors of symptom recurrence in mild persistent asthmatics who withdrew pharmacological therapy, as recommended in the step-down of international guidelines. PMID- 16253494 TI - Usefulness of the British Thoracic Society and the American College of Chest Physicians guidelines in predicting pleural drainage of non-purulent parapneumonic effusions. AB - AIM: To assess the value of the British Thoracic Society (BTS) and the American College of Chest Physicians (ACCP) guidelines to predict which patients with non purulent parapneumonic effusions (PPE) warrant chest tube drainage. METHODS: A retrospective chart review was performed on all patients who underwent thoracentesis because of a PPE over a 10-year period at a Spanish medical center. Classification of PPE as complicated (CPPE) or uncomplicated (UPPE) was based on the clinician's decision to insert a chest tube to resolve the effusion. Empyema was defined as pus in the pleural space. Data collected included patient demographics, size of the effusion, and microbiological and pleural fluid chemistries that might influence the physician's decision to place a chest tube. RESULTS: Of the 240 patients with PPE who entered the study, 85 had UPPE, 67 had CPPE, and 88 had empyema. Individual pleural fluid parameters, namely a pH<7.20, a glucose<40 mg/dL or <60 mg/dL, a LDH>1000 U/L or a positive culture had a relatively high specificity (from 78% for LDH to 94% for glucose<40 mg/dL), but low to moderate sensitivity (from 25% for culture to 73% for LDH) in predicting the need for chest tube placement in non-purulent PPE. While pleural fluid cultures performed poorly in discriminating UPPE from CPPE (likelihood ratio positive 1.7), effusion's size performed the best (likelihood ratio positive 5.7). BTS and ACCP guidelines yielded measures of sensitivity (98% and 97%, respectively), and negative likelihood ratio (0.03 and 0.05, respectively) for identifying a CPPE. CONCLUSIONS: Both guidelines have similar accuracy and perform satisfactorily in distinguishing CPPE from UPPE, albeit at an admissible cost of needlessly increasing chest tube drainage. PMID- 16253495 TI - Pulmonary rehabilitation for COPD. AB - Pulmonary rehabilitation is a therapeutic process, which entails taking a holistic approach to the welfare of the patient with chronic respiratory illness- most commonly chronic obstructive pulmonary disease (COPD). Pulmonary rehabilitation is considered essential throughout the lifetime management of patients with symptomatic chronic respiratory disease. It requires the coordinated action of a multidisciplinary healthcare team in order to deliver an individualised rehabilitation programme to best effect--incorporating multiple modalities, such as advice on smoking cessation, exercise training and patient self-management education, among others. As core components of pulmonary rehabilitation, exercise training and self-management education have been shown to be beneficial in improving health-related quality of life (HRQoL) in patients with chronic respiratory disease. Physical training can help to reduce the muscle de-conditioning that occurs when the activity of patients is restricted by their breathlessness and fatigue, and is often associated with an increase in patient HRQoL. HRQoL can also be improved by the use of self-management education, which is designed to provide the patient with the skills to manage the health consequences of their disease. In doing so, patients are better able to cope with disease symptoms, potentially leading to reduced healthcare costs. A great deal of research has been conducted to try and fully define which patients will benefit most from pulmonary rehabilitation. Although progress has been made, many questions remain as to the best means of delivering rehabilitation, particularly with respect to the optimum programme of physical training and patient self management education. PMID- 16253496 TI - Psychological outcomes of an outpatient pulmonary rehabilitation program in patients with chronic obstructive pulmonary disease. AB - STUDY/PRINCIPLES: The effects of an outpatient pulmonary rehabilitation program on psychological morbidity (anxiety and depressive symptoms) were examined in patients with chronic obstructive pulmonary disease (COPD). METHODS: The 26 rehabilitation patients with COPD were compared with 19 control patients with COPD similar in age, gender, COPD severity and other variables. Initial assessment included lung function testing, health status, exercise tolerance, dyspnea intensity and psychiatric interviews using Hamilton depression rating scale (HAM-D) and Hamilton anxiety rating scale (HAM-A). A pulmonary rehabilitation program was carried out during the following 2 months; psychiatric interviews and measurements of health status, exercise tolerance and dyspnea intensity were done again on completion of the study at 2 months. RESULTS: There was a decrease in HAM-A scores in the rehabilitation group and the decrease was statistically significant (P=0.010). On the contrary the HAM-A scores did not change in control group. The decrease in HAM-A scores in rehabilitation group was also statistically significant compared with the control group (P=0.042). There was no significant difference in HAM-D scores within the two groups and also there was no significant difference between the two groups in HAM-D scores. The health status, exercise tolerance and dyspnea intensity improved significantly in the rehabilitation group compared to the control group. CONCLUSION: This study shows that our outpatient rehabilitation program leads to a benefit in anxiety and depressive symptoms in COPD patients. The benefit was especially significant in anxiety symptoms. In addition to the improvement in psychological symptoms, the health status, exercise tolerance and dyspnea intensity were also significantly improved in COPD patients who underwent the rehabilitation program. This outpatient-based rehabilitation program was well accepted by the patients. The relatively simple design of the program makes it feasible independently of expensive equipment. PMID- 16253497 TI - Solid waste management in Macao: practices and challenges. AB - The rapid economic development and population growth in Macao have resulted in a large increase in refuse generated over the past decade. In 2003, the quantity of solid waste generated reached 249,255 tons, corresponding to 1.52 kg/day per capita. This figure has been gradually increasing. Domestic solid waste is the primary source of solid waste generation. The data showed that a considerable amount of the solid waste generated can be recycled and reutilized. Due to Macao's small geographic area and high cost of land, landfilling has the lowest priority for waste disposal. Solid waste incineration has been given a top priority over other waste disposal methods although it is much more expensive. In the last decade, more than 80% of the total waste in Macao was incinerated. However, the incineration capacity of the Macao Incineration Plant is going to reach its saturation earlier than expected. Waste minimization, the establishment of an effective waste collection and disposal fee system, and alternate ways dealing with the limited capacity of waste treatment facilities are regarded to be major challenges in the future. PMID- 16253498 TI - Acute severe asthma presenting in late pregnancy. AB - Asthma is the commonest pre-existing medical condition to complicate pregnancy. Acute severe asthma in pregnancy is rare, but poses difficult problems. In particular, the decision about when and where to deliver the fetus is complex, since maternal response to asthma treatment is unpredictable. We report the successful management of a parturient presenting with acute severe asthma at 37 weeks' gestation. The controversies involved and the importance of adopting a multi-disciplinary team approach to optimise maternal and neonatal outcomes are discussed. PMID- 16253500 TI - Response to the publication: EORTC guidelines for the use of erythropoietic proteins in anaemic patients with cancer. PMID- 16253501 TI - Why did the study fail? PMID- 16253503 TI - Telomerase inhibition, telomere shortening, cell growth suppression and induction of apoptosis by telomestatin in childhood neuroblastoma cells. AB - Neuroblastoma is a tumour derived from primitive cells of the sympathetic nervous system and is the most common extracranial solid tumour in childhood. Unfavourable tumours are characterised not only by structural changes, including 1p deletion and amplification of the MYCN proto-oncogene, but also by high telomerase activity. Telomeric G-rich single-stranded DNA can adopt in vitro an intramolecular quadruplex structure, which has been shown to inhibit telomerase activity. In this study, we examined telomestatin, a G-quadruplex interactive agent, for its ability to inhibit telomere maintenance of neuroblastoma cells. Telomere length was determined by the terminal restriction fragment method, telomerase activity was measured by a quantitative telomeric repeat amplification protocol, and the expression of human telomerase by quantitative real-time polymerase chain reaction (RT-PCR). Short-term treatment with telomestatin resulted in dose-dependent cytotoxicity and induction of apoptosis. Long-term treatment with telomestatin at non-cytotoxic, but still telomerase activity inhibiting, concentrations resulted in telomere shortening, growth arrest and induction of apoptosis. These results suggest that the effect of telomestatin is dose-dependent and at least 2-fold. Prolonged low-dose treatment with telomestatin limits the cellular lifespan of NB cells through disruption of telomere maintenance. PMID- 16253502 TI - Fludarabine phosphate-CVP in patients over 60 years of age with advanced, low grade and follicular lymphoma: a dose-finding study. AB - The aim of this study was to establish a safe and effective regimen of fludarabine phosphate, cyclophosphamide, vincristine and prednisone (F-CVP) as first-line treatment for elderly patients with advanced, low-grade non-Hodgkin's lymphoma. Twenty-three patients >60 years were assigned successively to eight treatment cycles (Dose level 1: low F, low CV [n=4]; 2A: high F, low CV [n=8]; 2B: low F, high CV [n=4]; 3: high F, high CV [n=7]). High and low levels were: F, 25 and 20mg/m(2), respectively (Days 1-5); C, 750 and 500 mg/m(2), respectively (Day 1); and V, 1.4 and 1mg/m(2), respectively (Day 1). Patients received P at 40 mg/m(2) on Days 1-5. Response was assessed after Cycles 2, 4, 6 and 8. At level 3, dose-limiting toxicity (opportunistic infections and neutropenia) became evident, particularly after Cycle 6. Further patients were recruited at Dose level 2A. All regimens proved effective, with an OR rate of 78% (65% CR), and 3 year survival of 65% (+/-10%). Among 18 responders, 51% were still in response at 3 and 5 years. The study shows that this combination therapy is highly effective. The addition of F to CVP at Dose level 2A was feasible and increased the CR rate, with good tolerability in elderly patients. PMID- 16253504 TI - Dydrogesterone in the reduction of recurrent spontaneous abortion. AB - One hundred and eighty women with a history of recurrent, unexplained spontaneous abortion (mean 3.5 abortions) were randomised to receive oral dydrogesterone (10 mg b.i.d.), intramuscular human chorionic gonadotrophin (hCG; 5000 IU every 4 days) or no additional treatment (controls). Treatment was started as soon as possible after confirmation of pregnancy and continued until the 12th gestational week. All women received standard supportive care. Abortions were significantly (p < or = 0.05) less common in the dydrogesterone group (13.4%) than in the control group (29%); there were no statistically significant differences between the hCG group and the control group. There were no differences between the groups with respect to pregnancy complications or congenital abnormalities. In conclusion, hormonal support with dydrogesterone can increase the chances of a successful pregnancy in women with a history of recurrent spontaneous abortion. PMID- 16253505 TI - Metaplastic carcinoma of the breast with neuroglial differentiation. AB - We present a case of a 34-year-old female who had a rapidly enlarging right breast mass. Biopsy results were consistent with a neuroblastic neoplasm of the breast. Neoadjuvant chemotherapy for a neuroblastoma was performed followed by a modified radical mastectomy was performed. Final pathology revealed small foci of poorly differentiated carcinoma randomly distributed within extensive areas of neuroglial differentiation. Such a lesion has not been described in the current literature to our knowledge, and we propose that this neoplasm is a metaplastic carcinoma with a novel heterogeneous differentiation. Subsequent therapy consisted of standard adjuvant breast cancer chemotherapy and radiation therapy. PMID- 16253506 TI - Leaky guts and lipid rafts. AB - The intestinal epithelium functions as a physical barrier separating luminal microorganisms from the underlying immune system. There is compelling evidence that several intestinal diseases are associated with the translocation of commensal bacteria across the epithelial barrier. Recent work has identified a novel mechanism by which normally non-invasive enteric bacteria breach the intestinal epithelium during periods of inflammation. PMID- 16253507 TI - Stochastic entropy QSAR for the in silico discovery of anticancer compounds: prediction, synthesis, and in vitro assay of new purine carbanucleosides. AB - A Markov model based QSAR is introduced for the rational selection of anticancer compounds. The model discriminates 90.3% of 226 structurally heterogeneous anticancer/non-anticancer compounds in training series. External validation series were used to validate the model; the 91.8% containing 85 compounds, not considered to fit the model, were correctly classified. The model developed is afterwards used in a simulation of a virtual search for anticancer compounds never considered either in training or in predicting series. The 87.7% of the 213 anticancer compounds used in this simulated search were correctly classified. The model also shows high values for specificity (0.89), sensitivity (0.91), and Mathews correlation coefficient (0.79). In addition, the present model compares better-to-similar with respect to other four models elsewhere reported if one takes into consideration 26 comparison parameters. Finally, we exemplify the use of the model in practice with the design of a new series of carbanucleosides. The compounds evaluated with the model were synthesized and experimentally assayed for their antitumor effects on the proliferation of murine leukemia cells (L1210/0) and human T-lymphocyte cells (CEM/0 and Molt4/C8). The more interesting activity was detected for the compound 5a with a predicted probability of 80.2% and IC(50) = 27.0, 27.2, and 29.4 microM, respectively, against the above mentioned cellular lines. These values are comparable to those for the control compound Ara-A. PMID- 16253508 TI - Influence of a Chinese crude drug on Ca2+ influx and efflux in rat visceral organs: investigation and evaluation by 45Ca. AB - The influences of a Chinese crude drug, Herba Epimedii, (HE), on Ca2+ influx and efflux in the isolated rat aorta and some visceral organs were evaluated by using 45Ca as a radioactive tracer. Additionally, its protective effect on myocardial ischemia was investigated in live animals. The results indicated that HE has significant influence on Ca2+ influx and efflux in the isolated rat aorta, heart, and kidney, in that it can markedly block 45Ca entering into cell and can facilitate efflux of intracellular Ca2+. However, among the three kinds of extracts from HE, the alkali extracts have the most obvious effect on calcium channels in visceral organs. Even if the alkali extracts are diluted by water for 10 times, the material still has a rather strong inhibition effect on calcium channels. Fortunately, the three kinds of extracts have favorable protective effect on myocardial ischemia induced by drugs or by the ligation of the coronary artery. This is consistent with the results about the Ca2+ influx and efflux obtained by isotope tracer technique, and implies that the Chinese crude drug has attractive potential for the treatment of heart, cerebrovascular and other diseases. PMID- 16253509 TI - Psychological outcome of patients following treatment of oral cancer and its relation with functional status and coping mechanisms. AB - BACKGROUND: Traditionally health-care providers have measured outcome of treatment of disease by focusing on tumour response and disease-free survival. However, it has become increasingly apparent that the behavioural and functional impact of treatment on the patient is important. This study investigates the psychological outcome and its relationship with functional status and coping mechanisms following treatment of oral cancer patients. MATERIAL AND METHODS: Sixty-eight patients were evaluated 6 months to 6 years after treatment (from October 1992 to October 1997) for oral cancer. The Hospital Anxiety and Depression Scale (HADS) was used for psychological evaluation, the University of Washington Quality of Life Questionnaire (UW-QOL) and The European Organisation for Research and Treatment of Cancer Questionnaire (EORTC QLQ-C30) for evaluating the head and neck specific and general functional status, respectively. Finally, the "Mental Adjustment to Cancer Questionnaire" (MAC-Q) was used for evaluation of coping mechanisms. RESULTS: The incidence of anxiety and/or depression was 25% and the socio-demographic and medical characteristics showed poor correlation with the psychological outcome in this study. The results indicated that there was a strong association between psychological outcome and head and neck specific and general quality-of-life (QOL) domains, and style of coping. The p-value was less than 0.01 for most of the domains and items of functional status and the effective coping style. CONCLUSION: Deteriorated functional status and ineffective coping strategies are strongly associated with poor psychological outcome in patients with oral cancer. PMID- 16253510 TI - Does avascular necrosis of the jaws in cancer patients only occur following treatment with bisphosphonates? AB - INTRODUCTION: In the last decade, bisphosphonates were regularly used to treat osteoporosis and bone pain from diseases such as metastatic breast cancer, multiple myeloma and Paget's disease. Currently, the influence of bisphosphonates in development of avascular osteonecrosis of the jaws has been recognized by various authors. In many cancer patients chemotherapy and medications like steroids have also to be applied. Agreement exists that these drugs can initiate vascular endothelial cell damage and accelerate disturbances in the microcirculation of the jaws possibly resulting in thrombosis of nutrient end arteries. The role of bisphosphonates in cancer patients with previously treated jaws has yet to be elucidated. PATIENTS: Four case reports of 'cancer' patients are described in whom osteonecrosis of the jaws was found. In two patients, the nitrogen-containing bisphosphonate zoledronic acid was prescribed for additional therapy of malignancy for a period of 45 up to 70 months. In another case, supportive treatment of breast cancer was offered using ibandronate. The fourth patient suffered avascular necrosis of the mandible without ever having taken bisphosphonates. In any case, revisional, as well as extended surgery has to be performed for osteonecrosis because neither conservative debridement nor antibiotic therapy have shown long term success, with or without bisphosphonates. No withdrawal of bisphosphonates was performed in view of the information on the direct correlation of total dosage and duration of drug intake to systemic incorporation and the long time for drug release. CONCLUSION: According to our observations, withdrawal of bisphosphonates is not recommended when necrosis of the jaws has occurred. PMID- 16253511 TI - Experimental study of mineralization in mandibular bone distraction with irradiation during the consolidation phase. AB - INTRODUCTION: This study aimed at assessing the effect of radiotherapy on regenerated bone mineralization and consolidation obtained by mandibular bone distraction. The planned application was a new immediate mandibular reconstruction technique in defects following surgery for oral cavity tumours requiring postoperative radiotherapy. MATERIAL AND METHODS: Ten sheep underwent bilateral mandibular bone distraction (control group). A second group of 6 sheep (study group) had bilateral mandibular bone distraction followed by irradiation on the 21st postoperative day. The animals were sacrificed on the 60th postoperative day. Radiographic and histomorphometric studies were performed. RESULTS: In the control group, 9 distraction sites out of 20 were consolidated. In the irradiated group, 9 out of 12 were consolidated. Histomorphometric analysis did not demonstrate any statistically significant difference between the osseous and cartilaginous samples, the trabecular thickness of the new bone, or the osteoid surface of basal bone in the control and test groups (p=0.126, 0.247, 0.792, 0.082). However, the osteoid surface of the regenerated bone in the test group was statistically smaller (p=0.017) than in the control group. CONCLUSION: In this experiment, radiotherapy did not hinder bone mineralization or consolidation following distraction of mandibular bone receiving irradiation on the 21st postoperative day. Bone distraction could be proposed for the repair of mandibular defects following surgery for oral tumours which require early postoperative radiotherapy. PMID- 16253512 TI - Possible problems of moulding the regenerate in mandibular distraction osteogenesis -- experimental aspects in a canine model. AB - INTRODUCTION: Moulding of the regenerate created by distraction osteogenesis has been shown clinically to be efficient and good enough so that for complex three dimensional deformities, final adjustments by moulding the regenerate may be part of the treatment plan. This study assessed possible drawbacks of moulding a regenerate, taking into consideration compressive and tensile forces acting simultaneously on the fresh callus. METHOD: Distraction osteogenesis in 15 Beagle mandibles was performed using custom made devices which allowed for lengthening as well as for angulation. After linear distraction of 10 mm, a defined 20 degrees angulation was performed in one step. The position of the fulcrum of the device allowed simultaneously compression and stretching of the regenerated bone. Effects on bone healing were assessed after 6 and 13 weeks of consolidation respectively and compared with a control group where only linear distraction was performed. RESULTS: Radiological and histological investigations demonstrated that no significant differences between the biological behaviour of the compressed and the stretched zones of the regenerate could be found. However, there were signs showing the more critical character of the stretched area. After 6 weeks of consolidation some specimens revealed delayed ossification of the stretched zone. Under stable conditions, this delay was compensated for after 13 weeks of consolidation and complete osseous healing occurred. CONCLUSION: Under stable conditions, a fresh regenerate can be moulded to a considerable extent without permanently endangering osseous healing. Nevertheless tensile forces acting on the regenerate should be minimized to prevent damage to the new bone. This can be achieved by overdistraction prior to callus moulding or by gradually changing the vector of distraction during the lengthening process. PMID- 16253513 TI - Growth factor expression following clinical mandibular distraction osteogenesis in humans and its comparison with existing animal studies. AB - AIM: Lengthening the mandible by distraction osteogenesis (DO) is nowadays a well recognized technique in maxillofacial surgery. In this study growth factor expression profiles were examined in biopsies taken from six patients undergoing mandibular DO and compared with findings from a sheep model for mandibular DO. STUDY DESIGN: In all patients (and sheep), the ascending ramus was distracted 10 15 mm at a rate of 1mm/day using an intraoral device. Biopsies were taken from the centre of the distraction zone 21 days after completion of distraction. Using standard immunohistochemical techniques, samples were stained for platelet derived growth factor (PDGF), transforming growth factor beta (TGF-beta), basic fibroblast growth factor (bFGF) and bone morphogenetic proteins-2, -4 and -7 (BMP 2, -4, -7), matrix metalloproteinases-1 and -3 (MMP-1, -3), the vascular endothelial growth factor (VEGF), a marker for endothelial cells (CD-31) and type IV collagen (Col IV). RESULTS: Positive staining for PDGF, bFGF, TGF-beta, BMP-2, -4, and -7 was noted in cells and matrix components. There was intense staining for MMP-1. Strong staining for CD-31 and COL IV was observed adjacent to vessels. VEGF staining was less specific. Similar findings were noted in the sheep model. CONCLUSION: Growth factor expression in the human distraction site is similar to that in the sheep model. PMID- 16253514 TI - Cytotoxic and antitumor activity of liposome-incorporated sclareol against cancer cell lines and human colon cancer xenografts. AB - The aim of this study was to design and prepare liposome-incorporated sclareol--a highly lipophilic natural product-to overcome its water insolubility and develop suitable formulations for in vivo administration. The bioactive labdane-type diterpene sclareol was incorporated into liposomes composed of egg phosphatidylcholine and dipalmitoylphosphatidylglycerol prepared by the thin-film hydration method followed by sonication. A formulation of egg phosphatidylcholine/dipalmitoylphosphatidylglycerol/sclareol (9:0.1:5 molar ratio) was developed and characterized. The lipid recovery and the sclareol to lipid molar ratio were measured using high-performance thin-layer chromatography/flame ionization detection. In vitro drug release was measured in supplemented RPMI-1640 at 37 degrees C. The liposomal and the free sclareol were initially tested in vitro for their activity against human cancer cell lines using the sulphorhodamine B assay. Liposomes incorporating sclareol at a drug to lipid molar ratio of 0:43, suggesting an incorporation efficiency of almost 80%, showed reduced growth rate of human colon cancer tumors (HCT116) developed in SCID mice, without any significant side effects. PMID- 16253515 TI - In situ structural characterization of phosphatidylcholines in brain tissue using MALDI-MS/MS. AB - Phosphatidylcholine (PC) is one of the most abundant classes of phospholipids and is a major component of membranes in biological systems. Recently, PCs have been detected by direct tissue analysis using MALDI-TOFMS. However, these studies did not allow for the structural characterization of PCs in tissue. In the current study, an in situ method for detection and structural analysis of PC species in brain tissue was developed using a MALDI-TOF/TOF mass spectrometer. Initial profiling of lipids in tissue is performed by MALDI-TOFMS, which allows for the assignment of PC species. However, to confirm the structure of the PC species detected in tissue, MALDI-MS/MS analysis was employed. In this work, protonated, sodiated, and potassiated PC species were detected in brain tissue using DHA matrix. MALDI-MS/MS analysis of these species yielded fragments that verified a phosphocholine head group, but did not supply any fragments that would permit the identification of acyl substituents. To obtain more structural information, lithium adducts of PC species were produced using DHA matrix dissolved in 100 mM lithium chloride. MALDI-MS/MS analysis of lithiated PC species produced fragments that allowed for the identification and positional assignment of acyl groups in PC species. PMID- 16253516 TI - New and automated MSn approaches for top-down identification of modified proteins. AB - An automated top-down approach including data-dependent MS(3) experiment for protein identification/characterization is described. A mixture of wild-type yeast proteins has been separated on-line using reverse-phase liquid chromatography and introduced into a hybrid linear ion trap (LTQ) Fourier transform ion cylclotron resonance (FTICR) mass spectrometer, where the most abundant molecular ions were automatically isolated and fragmented. The MS(2) spectra were interpreted by an automated algorithm and the resulting fragment mass values were uploaded to the ProSight PTM search engine to identify three yeast proteins, two of which were found to be modified. Subsequent MS(3) analyses pinpointed the location of these modifications. In addition, data-dependent MS(3) experiments were performed on standard proteins and wild-type yeast proteins using the stand alone linear trap mass spectrometer. Initially, the most abundant molecular ions underwent collisionally activated dissociation, followed by data dependent dissociation of only those MS(2) fragment ions for which a charge state could be automatically determined. The resulting spectra were processed to identify amino acid sequence tags in a robust fashion. New hybrid search modes utilized the MS(3) sequence tag and the absolute mass values of the MS(2) fragment ions to collectively provide unambiguous identification of the standard and wild-type yeast proteins from custom databases harboring a large number of post-translational modifications populated in a combinatorial fashion. PMID- 16253517 TI - Investigation of the presence of b ions in electron capture dissociation mass spectra. AB - Previously, we have indicated (Cooper, H.J., et al. Int. J. Mass Spectrom., 2003, 228, 723-728) that electron capture dissociation (ECD) of the doubly protonated peptides, Leu(4)-Sar-Leu(3)-Lys-OH, Leu(4)-Ala-Leu(3)-Lys-OH, Gly(4)-Sar-Gly(3) Lys-NH(2), and Gly(3)-Pro-Sar-Gly(3)-Lys-NH(2), results in abundant b ions, which derive from fragmentation of backbone amide bonds, a nonstandard fragmentation channel in ECD. The instrumental conditions were such that the possibility that collision-induced dissociation processes were contributing to the observed spectra was eliminated. In a separate study (Fung, Y.M.E., et al. Eur. J. Mass Spectrom., 2004, 10, 449-457. ECD of peptides Arg-(Gly)(n)-Xxx-(Gly)(n)-Arg, where Xxx is the amino acid of interest, did not result in b ions. The variation in ECD observed for strikingly similar peptides suggests that the nature of the charge carrier (Arg or Lys) is instrumental in governing the fragmentation channels. Here, we describe the ECD behavior of a suite of model peptides designed such that the nature and position of the charge carrier could be probed. The results suggest that the presence of b ions in ECD spectra is a consequence of both charge carrier and peptide structure. Possible mechanisms for the formation of b ions following electron capture are discussed. PMID- 16253518 TI - Origin and evolution of the Trk family of neurotrophic receptors. AB - Among the numerous tyrosine kinase receptors, those belonging to the Trk family are distinctively involved in the development of complex traits within the vertebrate nervous system. Until recently, the lack of a proper Nt/Trk system in invertebrates has lead to the belief that they were a vertebrate innovation. Recent data, however, have challenged the field, and proved that bona fide Trk receptors do exist in invertebrates. Here, we review and discuss the evolutionary history of the Trk receptor family, and draw a comprehensive scenario that situates the origin of the Nt/Trk signalling prior to the origin of vertebrates. Probably, a ProtoTrk receptor was invented by means of domain and exon shuffling from pieces of ancient genes, generating the unique combination of domains found in extant Trk receptors. It is suggestive to propose that subtle protein mutations, gene duplications, and co-options in particular territories of a primitive Nt/Trk system were instrumental to the development of a complex vertebrate nervous system. PMID- 16253519 TI - Characterization of the mechanical and neural components of spastic hypertonia with modified H reflex. AB - As the H reflex remains unable to assess mechanical changes intrinsic to a muscle, the aim of this study was to modify the H reflex techniques and to characterize the neural and mechanical components of muscle spasticity, relating the two components to clinical observations. Thirty-four patients featuring either a spinal-cord lesion (n=15) or stroke (n=19) and 23 neurologically normal subjects were recruited. Soleus H reflex and maximal M response (M(max)) were measured with electromyography and mechanomyography (MMG). The motoneuronal excitability was represented with the adjusted ratio of the H reflex to the M(max) (H/M(max)) and the ratio of the paired H reflexes (H(2)/H(1)). Muscle mechanical properties were characterized by the amplitude and median frequency of maximal M response recorded with MMG (MMG(Mmax)). The results showed that spastic patients exhibited a larger H/M(max), H(2)/H(1) and amplitude of MMG(Mmax) than the control group. H/M(max) and amplitude of MMG(Mmax) accounted for 55.7% of the variance in the Modified Ashworth Scale, the clinical hypertonia assessment. The amplitude of MMG(Mmax) correlated with functional impairments, as assessed with the Barthel index and Fugl-Meyer motor-assessment scale. It was concluded that spastic hypertonia involved an atypical increase in motoneuronal excitability and muscle mechanical properties, while impairment of functional performance and daily activity was attributable primarily to altered mechanical properties of a spastic muscle. PMID- 16253520 TI - Attention modulates hemispheric differences in functional connectivity: evidence from MEG recordings. AB - The present study examined intrahemispheric functional connectivity during rest and dichotic listening in 8 male and 9 female healthy young adults measured with magnetoencephalography (MEG). Generalized synchronization within the separate hemispheres was estimated by means of the synchronization likelihood that is sensitive to linear as well as non-linear coupling of MEG signals. We found higher functional intrahemispheric connectivity of frontal and temporal areas within the right as compared to the left hemisphere in the lower and higher theta band during rest and in the lower theta band during dichotic listening. In addition, higher synchronization in the lower theta band correlated with better task performance. In the upper alpha band, hemispheric differences in intrahemispheric connectivity of the frontal regions were found to be modulated by focused attention instructions. That is, attention to the right ear exaggerates the pattern of higher synchronization likelihood for the right frontal region, while attention to the left ear has an opposite effect. We found higher intrahemispheric connectivity in males compared to females as shown by higher synchronization in the lower alpha band. Taken together, our results reflect a physiological basis for functional hemispheric laterality and support the general assumption of sex differences in brain organization. Furthermore, in addition to studies that show that controlled attention processes modulate activation of the frontal areas, our study indicates that attention modulates ipsilateral functional connectivity in the frontal areas. This supports the idea of a supervisory role for the frontal cortex in attention processes. PMID- 16253521 TI - Gray matter reduction is correlated with white matter hyperintensity volume: a voxel-based morphometric study in a large epidemiological sample. AB - Both brain atrophy and T2-weighted white matter hyperintensities (WMH) are common findings in the brains of asymptomatic elderly individuals as well as in disease specific brains. The study of the relationship between these two salient features is therefore important. To investigate such a relationship, we performed a brain magnetic resonance imaging (MRI) study on 397 asymptomatic individuals aged between 60 and 64 years, who were recruited randomly from a large community sample. WMH were delineated on T2-weighted fluid attenuation inversion recovery (FLAIR) whole brain scans using an automated procedure. The results showed that gray matter reduction, subarachnoid CSF (SA-CSF) increase and lateral ventricular dilation were significantly correlated with WMH load. Deep white matter hyperintensity (DWMH) had significant correlation with all three global atrophy indices, but periventricular white matter hyperintensity (PVWMH) was correlated only with gray matter volume. Voxel-based morphometric (VBM) analysis showed that regional gray matter reduction correlated more closely with WMH load of the proximate region than with WMH elsewhere. The results suggest that WMH have a relationship with brain atrophy in middle age, although the study cannot determine which process, i.e. the development of WMH or atrophy, is primary. The study also demonstrates that DWMH has a more significant relationship with structural brain changes, and may therefore be more functionally relevant than PVWMH. Further delineation of this relationship needs a longitudinal study of the changes in both WMH and indices of brain atrophy. PMID- 16253522 TI - Enhancing BOLD response in the auditory system by neurophysiologically tuned fMRI sequence. AB - Auditory neuroscience has not tapped fMRI's full potential because of acoustic scanner noise emitted by the gradient switches of conventional echoplanar fMRI sequences. The scanner noise is pulsed, and auditory cortex is particularly sensitive to pulsed sounds. Current fMRI approaches to avoid stimulus-noise interactions are temporally inefficient. Since the sustained BOLD response to pulsed sounds decreases with repetition rate and becomes minimal with unpulsed sounds, we developed an fMRI sequence emitting continuous rather than pulsed gradient sound by implementing a novel quasi-continuous gradient switch pattern. Compared to conventional fMRI, continuous-sound fMRI reduced auditory cortex BOLD baseline and increased BOLD amplitude with graded sound stimuli, short sound events, and sounds as complex as orchestra music with preserved temporal resolution. Response in subcortical auditory nuclei was enhanced, but not the response to light in visual cortex. Finally, tonotopic mapping using continuous sound fMRI demonstrates that enhanced functional signal-to-noise in BOLD response translates into improved spatial separability of specific sound representations. PMID- 16253523 TI - NG2 expression regulates vascular morphology and function in human brain tumours. AB - Tumour angiogenesis is a tightly regulated process involving cross-talk between tumour cells and the host tissue. The underlying mechanisms that regulate such interactions remain largely unknown. NG2 is a transmembrane proteoglycan whose presence on transformed cells has been demonstrated to increase proliferation in vitro and angiogenesis in vivo. To study the effects of NG2 during tumour growth and progression, we engineered an NG2 positive human glioma cell line (U251-NG2) from parental NG2 negative cells (U251-WT) and implanted both cell types stereotactically into immunodeficient nude rat brains. The tumours were longitudinally monitored in vivo using multispectral MRI employing two differently sized contrast agents (Gd-DTPA-BMA and Gadomer) to assess vascular leakiness, vasogenic oedema, tumour volumes and necrosis. Comparisons of Gd-DTPA BMA and Gadomer revealed differences in their spatial distribution in the U251 NG2 and U251-WT tumours. The U251-NG2 tumours exhibited a higher leakiness of the larger molecular weight Gadomer and displayed a stronger vasogenic oedema (69.9 +/- 15.2, P = 0.018, compared to the controls (10.7 +/- 7.7). Moreover, immunohistochemistry and electron microscopy revealed that the U251-NG2 tumours had a higher microvascular density (11.81 +/- 0.54; P = 0.0010) compared to controls (5.76 +/- 0.87), with vessels that displayed larger gaps between the endothelial cells. Thus, tumour cells can regulate both the function and structure of the host-derived tumour vasculature through NG2 expression, suggesting a role for NG2 in the cross-talk between tumour-host compartments. PMID- 16253524 TI - Neural representations of two-digit numbers: a parametric fMRI study. AB - When participants are asked to decide which of a pair of two-digit Arabic numbers is larger, they compare units and decades, even when units are irrelevant. Typically, behavioral responses are slower when units are incongruent with the decade comparison (e.g. 81_26, because 8 > 2 but 1 < 6; unit-decade compatibility effect, [Nuerk, H.-C., Weger, U., Willmes, K. 2001. Decade breaks in the mental number line? Putting the tens and units back in different bins. Cognition 82, B25 B33.]). We defined parametric regressors to examine the effect of decade digit distance and unit distance-based compatibility processing on the fMRI signal to investigate the neural correlates of two-digit magnitude processing. Fourteen male right-handed volunteers (mean age = 27, range 21-38 years) took part in the study. In a rapid event-related design, participants had to decide which of two two-digit Arabic numbers was larger. Data were preprocessed and analyzed statistically in SPM2. Activation in the anterior portion of the right IPS was significantly modulated by compatibility-based unit distance processing. Furthermore, decade distance predicted an increase in the fMRI signal from cortex around the left IPS, right anterior and right posterior IPS. These results indicate that magnitude representations of unit-decade compatibility and decade distance are subserved by the intraparietal cortex and that the symbolic structure of the Arabic number system is an important determinant of multi-digit number magnitude processing. Implications of the present results in terms of general symbolic information processing are discussed. PMID- 16253525 TI - Human brain structures related to plantar vibrotactile stimulation: a functional magnetic resonance imaging study. AB - The purpose of this study was to investigate the sensorimotor cortex response to plantar vibrotactile stimulation using a newly developed MRI compatible vibration device. Ten healthy subjects (20-45 years) were investigated. Vibrotactile stimulation of the sole of the foot with a frequency of 50 Hz and a displacement of 1 mm was performed during fMRI (echo-planar imaging sequence at 1.5 T) using an MRI compatible moving magnet actuator that is able to produce vibration frequencies between 0 and 100 Hz and displacement amplitudes between 0 and 4 mm. The fMRI measurement during vibrotactile stimulation of the right foot revealed brain activation contralaterally within the primary sensorimotor cortex, bilaterally within the secondary somatosensory cortex, bilaterally within the superior temporal, inferior parietal, and posterior insular region, bilaterally within the anterior and posterior cingular gyrus, bilaterally within the thalamus and caudate nucleus, contralaterally within the lentiform nucleus, and bilaterally within the anterior and posterior cerebellar lobe. The advantages of the new MRI compatible vibration device include effective transmission of the stimulus and controlled vibration amplitudes, frequencies, and intensities. The results indicate that plantar vibration can be a suitable paradigm to observe activation within the sensorimotor network in fMRI. Furthermore, the method may be used to determine the optimal responsiveness of the individual sensorimotor network. PMID- 16253526 TI - Functional localization in the human cerebellum based on voxelwise statistical analysis: a study of 90 patients. AB - The aim of the present study was to examine somatotopy in the cerebellar cortex and a possible differential role of the cerebellar cortex and nuclei in functional outcome. Clinical findings and 3D MRI-based cerebellar lesions site were compared in a group of 90 patients with focal cerebellar lesion using International Cooperative Ataxia Rating Scale (ICARS) and voxel-based lesion symptom mapping (VLSM). Separate analysis was performed in patients with acute and chronic ischemic lesions (n=43) and patients with acute and chronic surgical lesions (n=47). Thirty-eight patients were included after resection of a cerebellar tumor in childhood or adolescence. The most significant lesion symptom correlations were observed in the subgroup with acute ischemic lesions. Limb ataxia was significantly correlated with lesions of the interposed (NI) and part of the dentate nuclei (ND), ataxia of posture and gait with lesions of the fastigial nuclei (NF) including NI. Correlations with cortical lesions were less significant and present in the superior cerebellum only. Upper limb ataxia was correlated with lesions of vermal, paravermal and hemispheral lobules IV-V and VI, lower limb ataxia with lesions of vermal, paravermal and hemispheral lobules III and VI, dysarthria with lesions of paravermal and hemispheral lobules V and VI and ataxia of posture and gait with lesions of vermal and paravermal lobules II, III and IV. In the subgroups with chronic focal lesions, similar correlations were observed with lesions of the cerebellar nuclei, but significantly less correlations with lesions of the cerebellar cortex. Functional localization based on VLSM backs findings in previous animal and functional brain images studies in healthy human subjects. The lesion site appears to be critical for motor recovery. Lesions affecting the cerebellar nuclei are not fully compensated at any age and independent of the pathology in humans. PMID- 16253527 TI - Differential components of sentence comprehension: beyond single word reading and memory. AB - A number of studies have used functional neuroimaging to examine the neural mechanisms of sentence comprehension; however, few fMRI studies have examined activation patterns associated with sentence comprehension after accounting for activation attributable to single-word-level tasks important for sentence comprehension. To investigate the patterns of activation associated with sentence comprehension after controlling for single word reading and maintaining single words in memory, 20 unimpaired adult readers completed a block design paradigm which included sentence comprehension, single word reading, and short-term memory (for words) tasks. Results indicated that, regardless of the aspect of sentence comprehension being controlled for, activation was observed in bilateral temporal lobes (left > right) as well as bilateral occipital lobes and middle frontal gyri. Additional findings showed that bilateral superior parietal lobe activation was greatest for short-term memory for words, while left anterior inferior frontal gyri activation (centered around Brodmann's area 47) was greatest for single word reading. Results suggest that temporal cortex (left > right) is a core region important for sentence comprehension beyond the short-term memory and semantic requirements inherent in processing sentences. PMID- 16253529 TI - Gastrointestinal parasitism reduces the plasma availability of doramectin in lambs. AB - A study was undertaken to investigate the effect of parasitism on plasma availability and pharmacokinetic behaviour of doramectin (DRM) in lambs. Fourteen parasitised grey face Suffolk lambs (26.9 +/- 1.5 kg bodyweight) were selected for the study. Seven pairs of lambs were allocated to two groups to obtain an approximately even weight distribution. Group I (non-parasitised) was pre-treated with three repeated administrations of 5 mg/kg fenbendazole to maintain a parasite free condition. In group II (parasitised), the lambs did not receive any anthelmintic treatment. After the 85-day pre-treatment period, both groups of animals were treated with DRM by subcutaneous (SC) injection in the shoulder area at 200 microg/kg. Throughout the experimental period, both groups were maintained together under similar feeding and management conditions. Blood samples were collected by jugular venepuncture at different set times between 0.5 h and 60 days post-treatment. After plasma extraction and derivatisation, samples were analysed by high performance liquid chromatography (HPLC) with fluorescence detection. A computerised kinetic analysis was performed and the data were compared using the Student's paired t test. The parent molecule was detected in plasma between 30 min and either day 20 (parasitised) or day 35 (non-parasitised) post-DRM treatment. The AUC values of the parasitised group (143.0 +/- 18 ng d/mL) were significantly lower (P<0.05) than those observed in the parasitically naive animals (229.6 +/- 21.7 ng d/mL). The mean residence time (MRT) in the parasitised group (3.4 +/- 0.3 days) was significantly shorter (P<0.05) than in the healthy group (6.6 +/- 0.6 days). Study results have shown that parasitic disease, through alteration in the body condition, can produce significant changes in the plasma disposition of DRM when administered SC to parasitised lambs. PMID- 16253528 TI - Bcl-xL overexpression restricts gamma-radiation-induced apoptosis. AB - Bcl-xL belongs to a family of proteins which inhibit apoptosis in a number of stimuli including ionizing radiation. To better understand the effects and mechanisms of Bcl-xL on the apoptosis of lymphocytes and provide experimental basis to treat immune injury induced by radiation, we used normal human lymphoblastoid AHH-1 cells that were engineered to overexpress Bcl-xL proteins. Our results showed that overexpressed Bcl-xL reduced time-dependent increase of apoptosis induced by ionizing radiation. Reactive oxygen species (ROS) generation and Bax protein expression in the transfected AHH1-Bcl-xL cells were also lower compared to parental AHH-1 cells. Unexpectedly, the fluorescence intensity of Rhodomine 123 (Rh 123) for measuring mitochondrial membrane potential (MMP) did not change at all detected time points. These results possess a vital significance for insights into a new strategy for gene therapy of radiation induced immune injury. PMID- 16253530 TI - Measurement of magnetic susceptibility and calculation of shape factor of NMR samples. AB - A method for accurate measurement of magnetic susceptibility and determination of the shape factor in an NMR tube is shown. The combination of accurate shape factor determination with susceptibility measurement leads to improved accuracy when measuring chemical shift. This is important for comparing samples in different solvents or under different conditions, such as temperature, solvent, and pH. PMID- 16253531 TI - Kinase inhibitor recognition by use of a multivariable QSAR model. AB - We have applied a retrosynthetic program to determine the scaffold and R-group chemical space seen within a library of known kinase inhibitors and non-kinase drug-like molecules. Comparison of the differences quickly revealed that kinase inhibitors are distinct in several chemical fragment and physical properties. We then applied these descriptors in a multivariable quantitative structure-activity relationship (QSAR) model with the goal to distinguish kinase inhibitors from non kinase drug-like molecules. This model is heuristic in that it was trained over a dataset of 258 known kinase inhibitors and 230 non-kinase drug molecules. The final model recognized 98% of the training set as being kinase inhibitors and had a false positive rate of 15%. This trait for false positives was accepted out of a desire to maintain diversity and not miss possible good kinase inhibitors for screening. The model was validated by reserving a portion of the datasets as test sets, which were not included in the QSAR model building stage. This was done repetitively for different percentiles of the total dataset population. It was seen that model recognition and false positive were only slightly damaged well down to a 70% reserve (30% dataset used for QSAR model training while 70% used for reserve test set). Beyond 70%, the QSAR models were inconsistent, signifying that the training sets were inadequately diverse to represent the greater reserve test sets. We applied this model to evaluate the commercial kinase libraries available from Asinex, BioFocus, ChemDiv and LifeChemicals to facilitate purchase decisions for compounds for HTS for lead compounds. We observed that there are significant differences in populations of recognizable kinase inhibitors across the vendors analyzed, with BioFocus showing the greatest population of kinase like molecules. PMID- 16253532 TI - Molecular mechanism of the photoperiodic response of gonads in birds and mammals. AB - Appropriate timing of various seasonal processes is crucial to the survival and reproductive success of animals living in temperate regions. When seasonally breeding animals are subjected to annual changes in day length, dramatic changes in neuroendocrine-gonadal activity take place. However, the molecular mechanism underlying the photoperiodic response of gonads remains unknown for all living organisms. It is well known that a circadian clock is somehow involved in the regulation of photoperiodism. Recently, rhythmic expression of circadian clock genes was observed in the mediobasal hypothalamus (MBH) of Japanese quail. The MBH is believed to be the center for photoperiodism. In addition, long-day induced hormone conversion of the prohormone thyroxine (T(4)) to the bioactive triiodothyronine (T(3)) by deiodinase in the MBH has been proven to be important to the photoperiodic response of the gonads. Although the regulating mechanism for the photoperiodic response of gonads in birds and mammals has long been considered to be quite different, the long-day-induced expression of the deiodinase gene in the hamster hypothalamus suggests the existence of a conserved regulatory mechanism in avian and mammalian photoperiodism. PMID- 16253533 TI - Enzymic analysis of NADPH metabolism in beta-lactam-producing Penicillium chrysogenum: presence of a mitochondrial NADPH dehydrogenase. AB - Based on assumed reaction network structures, NADPH availability has been proposed to be a key constraint in beta-lactam production by Penicillium chrysogenum. In this study, NADPH metabolism was investigated in glucose-limited chemostat cultures of an industrial P. chrysogenum strain. Enzyme assays confirmed the NADP(+)-specificity of the dehydrogenases of the pentose-phosphate pathway and the presence of NADP(+)-dependent isocitrate dehydrogenase. Pyruvate decarboxylase/NADP(+)-linked acetaldehyde dehydrogenase and NADP(+)-linked glyceraldehyde-3-phosphate dehydrogenase were not detected. Although the NADPH requirement of penicillin-G-producing chemostat cultures was calculated to be 1.4 1.6-fold higher than that of non-producing cultures, in vitro measured activities of the major NADPH-providing enzymes were the same. Isolated mitochondria showed high rates of antimycin A-sensitive respiration of NADPH, thus indicating the presence of a mitochondrial NADPH dehydrogenase that oxidises cytosolic NADPH. The presence of this enzyme in P. chrysogenum might have important implications for stoichiometric modelling of central carbon metabolism and beta-lactam production and may provide an interesting target for metabolic engineering. PMID- 16253534 TI - Immunocharacterization of the mucin-type proteins from the intracellular stage of Trypanosoma cruzi. AB - The surface of Trypanosoma cruzi is covered by different groups of mucins that are differentially expressed during the parasite life cycle. We have previously identified the major mucins from the bloodstream trypomastigote stage. Here, we present additional evidence that together with our previous observations allows for the identification of a second mucin group also expressed in the mammal dwelling stages, but predominant in the intracellular amastigote. These mucins are encoded by many genes, are mostly composed of tandem repeats and are highly conserved except for an exposed hypervariable (HV) N-terminal peptide. Antibodies against HV-peptides are restricted to approximately 50% of the chronically infected human population, are monospecific (i.e. directed towards a single HV), and display low-avidity. In contrast, immunization with a single HV-peptide triggers high-avidity, cross-reacting humoral responses against multiple HV sequences, but not against other T. cruzi surface antigens. The diversity present in the HV regions and the characteristics of the antibody response against them suggest a role of these molecules in eluding and/or modulating the mammalian host immune system. PMID- 16253535 TI - Chimeric epitopes delivered by polymeric synthetic linear peptides induce protective immunity to malaria. AB - Polymeric linear peptide chimeras (LPCs) that incorporate Plasmodium vivax promiscuous T cell epitopes and the P. falciparum circumsporozoite protein B cell epitope have been shown to induce a high level of immunogenicity and overcome genetic restriction when tested as vaccine immunogens in BALB/c mice. The present study evaluates the biological relevance of several LPCs using a well characterized rodent malaria model. Polymeric peptide constructs based on P. berghei and P. yoelii sequences, and orthologous to the human malaria sequences included in the original LPCs, were designed and tested for immunogenicity in mice of different H-2 haplotypes. We demonstrate that robust immune responses are induced and that peptides containing the orthologous rodent Plasmodium sequences exhibited similar immunogenic capabilities. Unique to this report, we show that LPCs can also prime MHC class I-restricted cytotoxic T lymphocytes (CTLs) and, most relevantly, that a peptide construct prototype incorporating single B, T and CTL epitopes induced protection against an experimental challenge with P. berghei or P. yoelii sporozoites. Collectively, these results suggest that polymeric polypeptide chimeras can be used as a platform to deliver subunit vaccines. PMID- 16253536 TI - Amyopathic dermatomyositis. AB - Amyopathic dermatomyositis (ADM) is a rare condition characterized by skin lesions suggestive of dermatomyositis (DM) without detectable muscle abnormalities after at least 2 years of follow-up. Pulmonary fibrosis is uncommon in patients with ADM. CASE REPORT: A 64-year-old woman presented with a 2 years and 6 months history of nondestructive polyarthritis. She had skin changes suggestive of DM, including a pink rash over the face, neck, and forearms; Gottron's papules over the metacarpophalangeal joints; and heliotrope edema of the eyelids. She reported no muscle symptoms. Findings were normal from muscle enzyme assays, electromyography, and muscle biopsies. A diagnosis of ADM was given. Early lung fibrosis was found. Investigations for a tumor were negative. DISCUSSION: ADM is a rare condition that may be an abortive form of DM with a favorable outcome and a lower risk of malignancy compared to classic DM. However, the development of pulmonary fibrosis may cloud the prognosis. PMID- 16253537 TI - The numerical simulation of osteophyte formation on the edge of the vertebral body using quantitative bone remodeling theory. AB - OBJECTIVES: To extend the quantitative prediction of the external shape of bone structure to the simulation of osteophyte formation on the edge of vertebral body. METHODS: The high-order nonlinear equation of bone remodeling was used to control osteophyte formation process. The idea of topology optimization in engineering was imported to allow the outgrowth of osteophyte. Osteophyte on the edge of a vertebral body in its mid-sagittal plane was simulated numerically. RESULTS: Osteophyte formation is an adaptive bone remodeling process in response to the progressive changes of mechanical environment, which were mainly caused by intervertebral disc degeneration. CONCLUSION: The numerical simulation in this paper extended the structural simulation based on the quantitative bone remodeling theory to bone morphological abnormity in orthopaedics, which can help to better understand the relationship between bone morphological abnormity and the mechanical environment. PMID- 16253538 TI - Evaluation of finger skin temperature in scleroderma patients cyclically treated with iloprost. AB - OBJECTIVES: To evaluate by computerized digital thermometry (CDT) the effect of cyclic iloprost infusions on finger skin temperature (FST) in scleroderma patients. METHODS: Twenty-two scleroderma outpatients (one man, 21 women, mean age 54.2 years) underwent three computerized digital thermometries, the first one 3 days before a 5-day cycle of iloprost infusion, the second the day of the fifth infusion soon after the end of the therapy and the third 1 month after. FST was evaluated under basal conditions, immediately after a cold test and after an 18 min recovery period. For comparison CDT was performed in 10 sex and age matched control subjects. RESULTS: FST was lower in scleroderma patients before iloprost infusion than in the control group either after cold test either at the end of the recovery period while no difference was found before the cold test. Soon after iloprost infusion and 1 month later the FST at the end of the recovery period increased from the pre-treatment value of 27.8 +/- 5.3 to 30.4 +/- 3.5 degrees C and to 30.0 +/- 4.5 degrees C, respectively (P < 0.05 for both values). No substantial differences were found concerning FST before and at the end of the cold test. CONCLUSION: Iloprost administration for 5 days allows to normalize the FST value after cold exposition not only immediately after the infusions but even at 1 month distance from the therapy. Despite its short half-life the effect of the drug on endothelium is protracted. PMID- 16253539 TI - Comparison of simultaneous continuous intracranial pressure (ICP) signals from a Codman and a Camino ICP sensor. AB - Simultaneous continuous intracranial pressure (ICP) signals from two different sensors were compared. Continuous ICP monitoring from two ICP sensors (i.e. Codman ICP MicroSensor; Johnson & Johnson, Raynham, MA and Camino OLM ICP; Camino Laboratories, San Diego, CA) placed within the brain parenchyma was performed in three patients within the intensive care unit (ICU) as part of routine management of severe subarachnoid hemorrhage. For each 6s time window mean ICP was computed, showing large differences in mean ICP values between the signals. Differences above 5 mmHg were observed in 13% of the 128,425 time windows derived from 214 h ICP recordings in these three patients. In one patient, mean ICP differed more than 10 mmHg in 23% of the time windows. Comparisons of 675,503 individual single pressure wave pairs of these 128,425 time windows revealed marginal differences in single wave amplitude (dP, i.e. pulse pressure) and latency (dT, i.e. rise time) values, suggesting that differences in mean ICP were caused by differences in baseline pressure. For the individual time windows were computed the mean wave amplitude and mean wave latency values according to a new algorithm. There were as well marginal differences between signals of mean wave amplitude and latency values. Thus, changes in baseline pressure affect mean ICP but not single pressure wave characteristics such as amplitude (dP) and (dT) latency values. PMID- 16253540 TI - The link between obesity and the built environment. Evidence from an ecological analysis of obesity and vehicle miles of travel in California. AB - AIMS: Obesity and physical inactivity are known to be risk factors for many chronic diseases including hypertension, coronary artery disease, diabetes, and cancer. We sought to explore the association between an indicator of transportation data (Vehicle Miles of Travel, VMT) at the county level as it relates to obesity and physical inactivity in California. METHODS: Data from the California Health Interview Survey 2001 (CHIS 2001), the US 2000 Census, and the California Department of Transportation were merged to examine ecological correlations between vehicle miles of travel, population density, commute time, and county indicators of obesity and physical inactivity. Obesity was measured by body mass index (BMI). Physical inactivity was based on self-reported behaviors including walking, bicycling, and moderate to vigorous activity. The unit of analysis was the county. Thirty-three counties in California with population size greater than 100,000 persons per county were retained in the analyses. RESULTS: CHIS 2001 statewide obesity prevalence ranged from 11.2% to 28.5% by county. Physical inactivity ranged from 13.4% to 35.7%. Daily vehicle miles of travel ranged from 3.3 million to 183.8 million per county. By rank bivariate correlation, obesity and physical inactivity were significantly associated (p<0.01). Furthermore, by rank analysis of variance, the highest mean rank obesity was associated with the highest rank of VMT (p<0.01). Similar rank patterns were observed between obesity and physical inactivity and commute time. Associations between VMT and physical inactivity were examined but failed to reach statistical significance. CONCLUSION: This analysis adds to the growing evidence supporting the association between VMT (a measure of automobile transportation) and obesity. An urban design characterized by over dependence on motorized transportation may be related to adverse health effects. PMID- 16253541 TI - Impact evaluation of a Dutch community intervention to improve health-related behaviour in deprived neighbourhoods. AB - This study investigates the impact of a 2-year community intervention on health related behaviour among adults aged 18-65 years living in deprived neighbourhoods in Eindhoven, The Netherlands. The intervention is evaluated in a community intervention trial with a quasi-experimental design in a longitudinal cohort survey (n=1926 and attrition rate: 31%) using postal questionnaires. In the 2 year implementation phase, more than 40 intervention activities were planned and delivered by intersectoral neighbourhood coalitions. Outcome measures were fruit consumption, vegetable consumption, physical activity, smoking, alcohol consumption and intermediate outcomes of behaviour (i.e. attitudes, self efficacy, awareness, knowledge and stages of change). The intervention demonstrated no evidence for an impact on vegetable consumption, physical activity, smoking and alcohol consumption and weak evidence for a small impact on (intermediate) outcomes of fruit consumption. PMID- 16253542 TI - Does living in a disadvantaged area entail limited opportunities to purchase fresh fruit and vegetables in terms of price, availability, and variety? Findings from the Brisbane Food Study. AB - Understanding the role environmental factors may play in the dietary behaviours of socioeconomic groups is relevant for efforts to reduce health inequalities. In contrast with international research, earlier findings from the Brisbane Food Study (BFS), Australia, found no relationship between area socioeconomic characteristics and dietary behaviours or location of food shops. This paper examines whether the price and availability of fruits and vegetables are socioeconomically patterned using data from the BFS. Fifty census collection districts were randomly sampled and all local (i.e. within 2.5 km) supermarkets, greengrocers and convenience stores were observed. Little or no differences in price and availability were found on the basis of area socioeconomic characteristics. PMID- 16253543 TI - Animal models of type 2 diabetes with reduced pancreatic beta-cell mass. AB - Type 2 diabetes is increasingly viewed as a disease of insulin deficiency due not only to intrinsic pancreatic beta-cell dysfunction but also to reduction of beta cell mass. It is likely that, in diabetes-prone subjects, the regulated beta-cell turnover that adapts cell mass to body's insulin requirements is impaired, presumably on a genetic basis. We still have a limited knowledge of how and when this derangement occurs and what might be the most effective therapeutic strategy to preserve beta-cell mass. The animal models of type 2 diabetes with reduced beta-cell mass described in this review can be extremely helpful (a) to have insight into the mechanisms underlying the defective growth or accelerated loss of beta-cells leading to the beta-cell mass reduction; (b) to investigate in prospective studies the mechanisms of compensatory adaptation and subsequent failure of a reduced beta-cell mass. Furthermore, these models are of invaluable importance to test the effectiveness of potential therapeutic agents that either stimulate beta-cell growth or inhibit beta-cell death. PMID- 16253544 TI - MRI findings of carcinosarcoma in the parotid gland. AB - True malignant mixed tumours of the salivary gland (carcinosarcoma) are rare tumours composed of both carcinomatous and sarcomatous components. They predominantly occur in the parotid gland. This report presents the clinical, histological and imaging findings of a carcinosarcoma arising in the deep lobe of the parotid. We present magnetic resonance imaging (MRI) findings of this tumour. MRI demonstrated heterogenous intermediate T2-W signal with a thick irregular enhancing rim was suggestive of a malignant lesion. PMID- 16253545 TI - From replication to cultivation: hot news from Haloarchaea. AB - Haloarchaea have developed into model organisms that are utilized to study many biological processes. Examples are the mechanisms of chromosome maintenance, gene expression and its regulation, protein export and degradation, and motility and sensing. In addition to the analysis of model species like Halobacterium salinarum and Haloferax volcanii, natural communities have been characterized. Halophilic Archaea were found in low-salt environments and are thus more widespread than previously thought. PMID- 16253546 TI - Flexible information storage in MUDR(II) EHR. AB - An important research task of the EuroMISE Centre is the applied research in the field of electronic health record (EHR) design including electronic medical guidelines and intelligent systems for data mining and decision support. The research in this field was inspired by several European projects. We have proposed a mathematical meta-description of a flexible information storage model based on the experience gathered in cooperation in those projects. In this model, we use two basic structures called a knowledge base and data files. We describe those two structures using the graph theory concepts. Furthermore, we use logical formulas to express conditions that should be valid. Additionally, we present a description of a global system architecture of a 3-tier EHR application with interfaces based on the latest technologies; predominately on Web Services, SOAP, XML, HTTP, CORBA, etc. According to our experience and test results gained from the MUDR EHR usage, we describe an open universal solution, which can be applied as the EHR kernel of hospital information systems. To realize this approach in a daily practice for health professionals we have started a co-operative project with clinical information systems developers. Within that project we are developing a new system for continual shared health care. PMID- 16253547 TI - Rat hepatic stellate cells become retinoid unresponsive during activation. AB - Hepatic stellate cells (HSC) play an essential role in fibrogenesis. Many stimuli cause HSC to activate, lose their Vitamin A and produce collagen. It is unclear whether Vitamin A loss causes activation, potentiates it or is simply an event in the cascade of activation changes. We determine if exogenous retinoids prevent the activation of freshly isolated rat HSC activated by plating on plastic. We also determine if retinoids: (1) reverse HSC activation; (2) maintain/restore HSC intracellular retinoid levels; (3) maintain expression of HSC nuclear receptors for retinoic acid (RAR) in HSC that are becoming activated or are chronically activated. Markers of activation in freshly isolated HSC were decreased by either retinol or retinoic acid without increases in HSC retinoid concentration. mRNA levels for RAR-alpha, RAR-beta and RAR-gamma, the nuclear receptors for retinoic acid, decreased during activation of freshly isolated HSC even with retinoid supplementation. RAR-alpha, RAR-beta and RAR-gamma mRNA and RAR-beta protein was undetectable in chronically activated HSC and remained absent after retinoic acid supplementation. Activation markers in chronically activated HSC were only slightly decreased after retinoid exposure. We conclude that exposure of HSC to extracellular retinoids diminishes some markers of activation but does not prevent HSC activation. PMID- 16253548 TI - Increased levels of HIV RNA detected in samples with viral loads close to the detection limit collected in Plasma Preparation Tubes (PPT). AB - BACKGROUND: The accurate and reliable quantification of HIV RNA is an essential part of the management of HIV infected individuals, and elucidation of factors that may affect HIV RNA measurements, such as the use of Vacutainer Plasma Preparation Tubes (PPT), is crucial. OBJECTIVES: The objective of this study was to determine if plasma samples with viral loads close to the lower limit of the dynamic range of the assay collected in PPT tubes had increased levels of HIV RNA as compared to samples collected in standard EDTA tubes. STUDY DESIGN: HIV RNA levels were compared in 112 paired plasma samples collected in PPT and standard EDTA tubes. All samples had been frozen prior to testing. RESULTS: Discrepancies between PPT and EDTA tubes did not occur for samples with high viral loads. However, in samples with viral loads close to the lower limit of the dynamic range, levels of HIV RNA detected were higher in a large proportion of PPT as compared to the corresponding EDTA plasma samples. Forty percent of plasma pairs had no detectable HIV RNA in the EDTA aliquot, but had low levels of HIV RNA in the corresponding PPT aliquot. CONCLUSIONS: This prospective study underlines the need for cautious interpretation of small transient viral load changes in samples with values close to the detection limit. PMID- 16253549 TI - Is there a role of clinical neurophysiology in primary headache? PMID- 16253550 TI - Focal cortical high-frequency oscillations trigger epileptic spasms: confirmation by digital video subdural EEG. AB - OBJECTIVE: To localize high-frequency oscillations (HFOs) on the cortex during epileptic spasms using video subdural EEG and Multiple Band Frequency Analysis (MBFA). METHODS: Using video subdural EEG sampled at 1 kHz, we studied a 14-year old boy with asymmetric epileptic spasms of possible left frontal origin. We identified HFOs, then analyzed and localized their distributions by MBFA. We correlated HFO distribution to clinical spasm intensity. RESULTS: Ictal subdural EEG recorded HFOs at 60-150 Hz lasting 0.3-4 s. MBFA showed extensive but noncontiguous distribution of HFOs predominantly over the left frontal and temporal regions. HFOs began and became quasiperiodic before manifestation of clinical spasms. As clinical spasms intensified, HFOs persisted in regions where they initiated subclinically but were of higher frequency and greater power than HFOs in other regions. We performed cortical resections over the left frontal and temporal regions with predominant HFOs. Six months after surgery, the patient remained seizure free. CONCLUSIONS: HFOs were present over the ictal onset zone during epileptic spasms. Periodic spasms in this patient had the characteristics of partial seizures. SIGNIFICANCE: We show that HFOs occurred over the cerebral cortex during epileptic spasms, and we suggest that these focal cortical HFOs triggered the spasms. PMID- 16253551 TI - Source localization of mesial temporal interictal epileptiform discharges: correlation with intracranial foramen ovale electrode recordings. AB - OBJECTIVE: We have investigated the localization accuracy of low-resolution electromagnetic tomography (LORETA) for mesial temporal interictal epileptiform discharges (IED) on a statistical basis by using clinical electroencephalographic (EEG) data of simultaneous scalp and intracranial foramen ovale (FO) electrode recordings. METHODS: We retrospectively analyzed the IED of 15 patients who underwent presurgical assessment for intractable temporal lobe epilepsy. All patients have subsequently undergone amygdalohippocampectomy. The scalp signals were averaged time-locked to the peak activity in bilateral 10-contact FO electrode recordings. Source modeling was carried out by using statistical non parametric mapping (SNPM) of LORETA values and by calculating raw LORETA values of averaged IED. The results were compared to intracranial data obtained from FO electrode recordings. RESULTS: Two thousand six hundred and fifteen discharges could be attributed to 19 different patterns of intracranial mesial temporal IED. SNPM of LORETA revealed confined ipsilateral mesial temporal solutions for 14 (73.7%) and no significant solutions for five (26.3%) of these patterns. Raw LORETA current density distributions of the 19 averaged IED patterns revealed ipsilateral basal to lateral temporal solutions for the 14 IED patterns with a sufficient signal to noise ratio (SNR), but spurious results for those five IED with a low SNR. CONCLUSIONS: SNPM of LORETA but not LORETA analysis of averaged IED patterns accurately localizes the source generators of mesial temporal IEDs. SIGNIFICANCE: SNPM of raw LORETA values might be appropriate for localizing restricted mesial temporal lobe sources. PMID- 16253552 TI - Extrauterine life duration and ontogenic EEG parameters in preterm newborns with and without major ultrasound brain lesions. AB - OBJECTIVE: To determine whether extrauterine life duration is associated with different ontogenic electroencephalographic (EEG) patterns in premature infants compared with neonates born at later gestational ages (GAs). METHODS: We included 97 preterm infants of 24-36 weeks GA. We performed neurosonography and EEG recordings at week 1 and 2 of postnatal life, then every two weeks until term conceptional age (CA). We analyzed background EEG and sleep states and compared them at matched CA. RESULTS: In the group of infants without major ultrasound lesion (MUL), those <29 weeks GA showed increased sawtooth at 32-36 weeks CA and delta brushes from 34 to 40 weeks CA compared to infants > or =29 weeks GA. Infants with MUL compared to those without MUL showed significantly longer interburst intervals, shorter minimum burst duration at 2 and 4 weeks postnatal life, and higher incidence of sawtooth and delta brushes from 34 weeks CA to term. In addition, those of <29 weeks GA had higher rates of indeterminate sleep until 32 weeks CA. The type of MUL most frequently associated with dysmature EEG patterns was persistent ventricular dilatation. CONCLUSIONS: Premature infants without MUL showed prolonged dysmature EEG patterns as from early CAs (> or =29 weeks). This finding was more evident in those with MUL. SIGNIFICANCE: Extrauterine EEG development in prematures may differ from that of babies born at a later GA. PMID- 16253553 TI - Dynamics of the EEG slow-wave synchronization during sleep. AB - OBJECTIVE: To study the dynamics of spatial synchronization of the slow-wave activity recorded from different scalp electrodes during sleep in healthy normal controls. METHODS: We characterized the different levels of EEG synchronization during sleep (in the 0.25-2.5 Hz band) of five healthy subjects by means of the synchronization likelihood (SL) algorithm and analyzed its long-range temporal correlations by means of the detrended fluctuation analysis (DFA). RESULTS: We found higher levels of interregional synchronization during 'cyclic alternating pattern' (CAP) sleep than during nonCAP with a small but significant difference between its A and B phases. SL during CAP showed fluctuations probably corresponding to the single EEG slow-wave elements. DFA showed the presence of two linear scaling regions in the double-logarithmic plot of the fluctuations of SL level as a function of time scale. This indicates the presence of a characteristic time scale in the underlying dynamics which was very stable among the different subjects (1.23-1.33 s). We also computed the DFA exponent of the two scaling regions; the first, with values approximately 1.5, corresponded to fluctuations with period 0.09-0.75 s and the second, with values approximately 1, corresponded to fluctuations with period 1.5-24.0 s. Only the first exponent showed different values during the different sleep stages. CONCLUSIONS: All these results indicate a different role for each sleep stage and CAP condition in the EEG synchronization processes of sleep which show a complex time structure correlated with its neurophysiological mechanisms. SIGNIFICANCE: Very slow oscillations in spatial EEG synchronization might play a critical role in the long-range temporal EEG correlations during sleep which might be the chain of events responsible for the maintenance and correct complex development of sleep structure during the night. PMID- 16253555 TI - Human EEG gamma oscillations in neuropsychiatric disorders. AB - Due to their small amplitude, the importance of high-frequency EEG oscillations with respect to cognitive functions and disorders is often underestimated as compared to slower oscillations. This article reviews the literature on the alterations of gamma oscillations (about 30-80 Hz) during the course of neuropsychiatric disorders and relates them to a model for the functional role of these oscillations for memory matching. The synchronous firing of neurons in the gamma-band has been proposed to bind multiple features of an object, which are coded in a distributed manner in the brain, and is modulated by cognitive processes such as attention and memory. In certain neuropsychiatric disorders the gamma activity shows significant changes. In schizophrenic patients, negative symptoms correlate with a decrease of gamma responses, whereas a significant increase in gamma amplitudes is observed during positive symptoms such as hallucinations. A reduction is also observed in Alzheimer's Disease (AD), whereas an increase is found in epileptic patients, probably reflecting both cortical excitation and perceptual distortions such as deja vu phenomena frequently observed in epilepsy. ADHD patients also exhibit increased gamma amplitudes. A hypothesis of a gamma axis of these disorders mainly based on the significance of gamma oscillations for memory matching is formulated. PMID- 16253556 TI - The measure of randomness by leave-one-out prediction error in the analysis of EEG after laser painful stimulation in healthy subjects and migraine patients. AB - OBJECTIVE: We aimed to perform a quantitative analysis of event-related modulation of EEG activity, resulting from a not-warned and a warned paradigm of painful laser stimulation, in migraine patients and controls, by the use of a novel analysis, based upon a parametric approach to measure predictability of short and noisy time series. METHODS: Ten migraine patients were evaluated during the not-symptomatic phase and compared to seven age and sex matched controls. The dorsum of the right hand and the right supraorbital zone were stimulated by a painful CO(2) laser, in presence or in absence of a visual warning stimulus. An analysis time of 1s after the stimulus was submitted to a time-frequency analysis by a complex Morlet wavelet and to a cross-correlation analysis, in order to detect the development of EEG changes and the most activated cortical regions. A parametric approach to measure predictability of short and noisy time series was applied, where time series were modeled by leave-one-out (LOO) error. RESULTS: The averaged laser-evoked potentials features were similar between the two groups in the alerted and not alerted condition. A strong reset of the beta rhythms after the painful stimuli was seen for three groups of electrodes along the midline in patients and controls: the predictability of the series induced by the laser stimulus changed very differently in controls and patients. The separation was more evident after the warning signal, leading to a separation with P-values of 0.0046 for both the hand and the face. DISCUSSION: As painful stimulus causes organization of the local activity in cortex, EEG series become more predictable after stimulation. This phenomenon was less evident in migraine, as a sign of an inadequate cortical reactivity to pain. SIGNIFICANCE: The LOO method enabled to show in migraine subtle changes in the cortical response to pain. PMID- 16253557 TI - Differential modulations of intracortical neural circuits between two intrinsic hand muscles. AB - OBJECTIVE: To investigate whether the intracortical inhibitory (ICI) and facilitatory (ICF) circuits in the primary motor cortex between the first dorsal interosseous (FDI) and abductor digiti minimi (ADM) muscles are modulated differently. METHODS: We conducted paired-pulse transcranial magnetic stimulation in combination with different current directions (anterior-medially: AM, and posterior-laterally: PL) under relaxed and active muscle conditions with interstimulus intervals (ISIs) between 2 and 16 ms. RESULTS: In both muscle conditions, the conditioned motor-evoked potential (MEP) responses obtained with the AM current direction (preferentially eliciting early I-waves) were similar between the two muscles at all ISIs, but the MEP responses obtained with the PL current direction (preferentially eliciting late I-waves) were different between FDI and ADM muscles, in that the conditioned MEP responses in FDI muscle were inhibited at all ISIs under both muscle conditions, whereas those in ADM muscle were suppressed at only short ISIs (2-4 ms). CONCLUSIONS: These results indicate that the inhibitory connections operating for the corticospinal tract neurons in FDI muscle are more potent, and, conversely, that those in ADM muscle are weaker. SIGNIFICANCE: The different modulations of ICI circuits between FDI and ADM muscles is an important neural mechanism which may contribute to different functional demands (finger dexterity). PMID- 16253559 TI - The search for a tuberculosis vaccine: an elusive quest? AB - The precise mechanisms of protective immunity and pathogenesis of tuberculosis (TB) are poorly understood. Yet, many efforts are underway to develop new, more effective vaccines against Mycobacterium tuberculosis. This development appears to be predicated upon the assumption that natural acquired immunity to TB exists. In this paper we review the evidence for such immunity, explore the hypothesis that disease progression is due to some, often transient, immunological dysfunction, and discuss its relevance for vaccine development. Several mechanisms for such an immune dysfunction are proposed, including the recently suggested involvement of viral co-infections. Developing vaccines for such co infections may be a new challenge for TB control. PMID- 16253558 TI - Vaccine-induced cytokine responses in a guinea pig model of pulmonary tuberculosis. AB - Guinea pigs exposed to very small numbers of virulent tubercle bacilli by the respiratory route develop a disease which mimics many of the important features of the pathogenesis of human tuberculosis (TB), including the expression of strong protective immunity following vaccination with BCG. In order to elucidate the precise immunological mechanisms of vaccine-induced resistance in this model, both mRNA and protein assays for several guinea pig cytokines and chemokines have been developed. The coordinated expression of cytokine and chemokine mRNA and protein was examined in various leukocyte populations and in inflammatory cells and fluid collected following the induction of tuberculous pleurisy in BCG vaccinated guinea pigs. Real-time RT-PCR assays revealed that the mRNA levels for IFNgamma, TNFalpha, and IL-8 rose over the first few days of TB pleuritis and then declined over the 9 days of the study. Injection of anti-TGFbeta on day 8 following pleurisy induction resulted in significant changes in cytokine mRNA levels and PPD-induced proliferation in pleural effusion lymphocytes taken 24h later. BCG vaccination induced significantly higher levels of bioactive TNFalpha protein in the supernatants of alveolar, peritoneal and splenic cells from BCG vaccinated guinea pigs cultured in the presence of attenuated or virulent mycobacteria. In sharp contrast, following virulent challenge, all three cell types from BCG-vaccinated guinea pigs produced significantly less TNFalpha. Thus, BCG vaccination appears to modulate the potentially harmful effects of TNFalpha in this model of pulmonary TB. Levels of mRNA for IL-12p40 were upregulated by exposure of infected and uninfected macrophages to recombinant guinea pig (rgp)TNFalpha. The intracellular survival of mycobacteria was enhanced when endogeous TNFalpha activity was neutralized with anti-rgpTNFalpha antiserum. rgp RANTES (CCL5) upregulated mRNA levels for TNFalpha, IL-1beta, MCP-1 (CCL2), and IL-8 (CXCL8) in alveolar and peritoneal macrophages. These results illustrate the profound effects of prior vaccination with BCG on the cytokine and chemokine responses of distinct cell populations in the guinea pig following exposure to attenuated and virulent strains of M. tuberculosis. PMID- 16253560 TI - Bovine tuberculosis in India: potential basis for zoonosis. AB - Our laboratory has designed a specific nested-PCR (N-PCR) assay, based on the hupB gene of Mycobacterium tuberculosis (Rv2986c) and Mycobacterium bovis (Mb3010c) as a method to differentiate these closely related species. The present paper deciphers the utility of this assay for identification of pathogenic Mycobacteria in clinical samples. Extra-pulmonary clinical samples obtained from cattle and humans were investigated. Pre-dominance of M. tuberculosis (15.7%) and M. bovis (26.8%) was seen in humans and cattle, respectively. However, more importantly, both mycobacterial pathogens (mixed infection) were identified in a number of samples. In humans 8.7% of the samples and 35.7% in cattle were classified as mixed infection. The detection of mixed infection with the mycobacterial pathogenic duo in humans and bovines denotes the prospect of potential transmission of these pathogens from humans to cattle (zoonosis) and vice versa (reverse zoonosis). PMID- 16253561 TI - Recombinant and synthetic peptides to identify Mycobacterium tuberculosis antigens and epitopes of diagnostic and vaccine relevance. AB - The failures of Bacillus Calmette Guerin (BCG) as a vaccine and purified protein derivative as a diagnostic reagent in controlling the worldwide prevalence of tuberculosis (TB) have accelerated the research to identify Mycobacterium tuberculosis-specific antigens that could be useful as new vaccines and diagnostic reagents against TB. In the recent years, the comparative analyses of M. tuberculosis genome with the genomes of other mycobacteria have led to the identification of several genomic regions of M. tuberculosis that are deleted in BCG and other mycobacteria. These deleted regions (RDs) are predicted to encode over 100 proteins. If found immunologically reactive, the proteins encoded by M. tuberculosis-specific RDs could be useful in the specific diagnosis of TB and developing new vaccines. Among the approaches available for immunological characterization of the predicted M. tuberculosis-specific proteins are the evaluations of recombinant proteins and/or overlapping synthetic peptides, covering the sequence of each protein, for antibody and/or Th1 cell reactivity. These approaches have resulted into the identification of several antigenic proteins of M. tuberculosis encoded by genes located in RD1 with potentials in specific diagnosis of TB in low endemic areas and/or development of new vaccines, e.g. ORF14, ESAT6, CFP10, PE, PPE proteins, etc. In addition, prediction programs to identify peptides that could bind several HLA molecules, and presented to T cells in a promiscuous manner, have been developed. These programs have been used, on a limited scale, to identify the promiscuous peptides encoded by the genes spanning the M. tuberculosis-specific sequence. The promiscuous antigens/peptides recognized by T-cells in cell mediated immunity assays may have potentials in developing peptide-based vaccines and diagnostic reagents against TB. PMID- 16253563 TI - Superior virulence of Mycobacterium bovis over Mycobacterium tuberculosis (Mtb) for Mtb-resistant and Mtb-susceptible mice is manifest as an ability to cause extrapulmonary disease. AB - Mice of a Mycobacterium tuberculosis-resistant (BALB/c) and of a M. tuberculosis susceptible (DBA/2) strain proved considerably more susceptible, and equally so, to infection with Mycobacterium bovis than with M. tuberculosis when infection was initiated via the iv route. Infection with M. tuberculosis was eventually controlled at an approximately stationary level in the lungs, livers, spleens and kidneys of BALB/c mice, and in all of these organs except the lungs in DBA/2 mice. M. tuberculosis-infected DBA/2 mice died with a much shorter median survival time (MST) than M. tuberculosis-infected BALB/c mice. By contrast, infection with M. bovis killed mice of both strains with the same and much shorter MST. Unexpectedly, M. bovis caused progressive infection and pathology in the livers of BALB/c mice, but not in this organ in DBA/2 mice. More importantly, this pathogen caused progressive infection and infection-induced pathology in the kidneys and adrenal glands of both strains of mice. It is proposed that disease of the adrenal glands might serve to explain why M. bovis caused mice of both strains to die with the same much shorter MST. PMID- 16253562 TI - Revised national TB control programme in India. AB - The Revised National TB Control Programme (RNTCP), an application in India of the Directly Observed Treatment, Short Course (DOTS) strategy to control TB is being implemented in the country since 1997. The goal of the RNTCP is to decrease mortality and morbidity due to TB and cut transmission of infection until TB ceases to be a major public health problem in India Since 1999, achievements of the RNTCP have largely determined the global DOTS progress. It has been cited as the fastest expansion in the history of DOTS and in terms of patients being treated, it is the largest programme in the world. More than 50-fold expansion in RNTCP coverage has occurred since 1998 thus making DOTS accessible to more than 83% of the country's population. Quality of services has been maintained during this rapid expansion. As a result, the proportion of sputum positive cases confirmed in the laboratory is on par with international standards. By September 2005, the programme has initiated more than 3.5 million patients on treatment, thus saving over 600,000 additional lives. In 2003, new sputum positive case detection rate of 69% was achieved against target of at least 70% and treatment success rate of 86% has been achieved above the target of 85% Aggressive steps are being taken to meet global TB control targets by covering the entire country with RNTCP by 2005. Despite these achievements, there are many challenges for the RNTCP. Implementing DOTS in a diverse and large country, maintaining the quality of services during rapid expansion phase, decentralization of programme management to the states and, widening the reach of the programme to reach all sections of the society are some of the major challenges. PMID- 16253564 TI - The unusual chaperonins of Mycobacterium tuberculosis. AB - Heat shock proteins (Hsps), also known as molecular chaperones, are a diverse set of proteins that mediate the correct folding, assembly, transport and degradation of other proteins. In addition, Hsps have been shown to play a variety of important roles in immunity, thereby representing prominent antigens in the humoral and cellular immune response. Chaperonins form a sub-group of molecular chaperones that are found in all domains of life. Chaperonins in all bacteria are encoded by the essential groEL and groES genes, also called cpn60 and cpn10 arranged on the bicistronic groESL operon. Interestingly, Mycobacterium tuberculosis contains two copies of the cpn60 genes. The existence of a duplicate set of cpn60 genes in M. tuberculosis, however, has been perplexing. Cpn10 and Cpn60s of M. tuberculosis have been shown to be highly antigenic in nature, eliciting strong B- and T-cell immune responses. Recent work has shown intriguing structural, biochemical and signaling properties of the M. tuberculosis chaperonins. This review details the recent developments in the study of the M. tuberculosis chaperonins. PMID- 16253565 TI - Double-stranded RNA induces production of RANTES and IL-8 by human nasal fibroblasts. AB - Double-stranded RNA (dsRNA) and the viral RNA mimic, polyinosine-polycytidylic acid (poly(I:C)), are recognized by toll-like receptor 3 (TLR3) that mediates the innate immune response to viral infections. In this study, we investigated the effects of poly(I:C) on the production of chemokines (IL-8, RANTES, and eotaxin), Type I IFNs (IFNalpha and IFNbeta), Th1-cytokines (IL-12 and IFNgamma), and pro inflammatory cytokines (TNF-alpha and IL-1beta) by human nasal mucosa-derived fibroblasts. Human nasal fibroblasts were treated with poly(I:C), and levels of cytokines and chemokines were measured by ELISA. Incubation with poly(I:C) significantly enhanced the secretion of RANTES and IL-8. However, eotaxin, IL 1beta, TNF-alpha, IFNalpha, IFNgamma, and IL-12 were not secreted from nasal fibroblasts stimulated with poly(I:C). The JNK inhibitor SP600125 and the PI3 kinase inhibitor LY294002 significantly blocked the poly(I:C)-induced release of RANTES and IL-8, whereas the p38 MAP kinase inhibitor SB203580 suppressed poly(I:C)-induced secretion of IL-8, but not RANTES. Nasal fibroblasts play an important role in initiating antiviral responses and inflammation of the nasal cavity by producing chemokines leading to enhanced inflammatory cell recruitment. PMID- 16253566 TI - Risk factors for psychogenic nonepileptic seizures in children and adolescents with epilepsy. AB - There is evidence that psychogenic nonepileptic seizures (PNES) remain underdiagnosed, especially in children and adolescents. Diagnosis of such events is even more difficult in patients that do have epilepsy, leading to delayed diagnosis and treatment and, consequently, iatrogenic complications. This study aimed to evaluate possible risk factors in children with epilepsy who had PNES. Seizures and epileptic syndromes were classified according to International League Against Epilepsy guidelines. Patients were evaluated with a structured psychiatric anamnesis and classified according to the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition; Classification of Mental and Behavioral Disorders: Diagnostic Criteria for Research; and Schedule for Affective Disorders and Schizophrenia for School-Age Children--Epidemiological Version. Risk factors such as head trauma, physical, sexual and psychological abuse, and psychiatric diagnoses, among others, were investigated. Family history of epilepsy and psychiatric illness were detected by review of medical records and/or follow-up interviews. Gender was not a predictive factor, and although older children had a higher risk for PNES, younger children also presented truly psychogenic events mimicking epileptic seizures. The most common associated psychiatric diagnosis was depression. Family histories for epilepsy and psychiatric illness were a frequent finding. An inadequate family environment was more common than sexual or physical abuse. Current knowledge obtained from adults with PNES has been used to understand children with PNES. However, this study of children with epilepsy revealed some similarities and many differences. These features may help to identify predictive factors in a population in need of adequate diagnosis of and therapy for this long-lasting pathology. PMID- 16253567 TI - Social cognition and epilepsy surgery. AB - Human social behavior depends on a set of perceptive, mnemonic, and interpretive abilities that together may be termed social cognition. Lesion and functional imaging studies of social cognitive functions implicate the temporal lobes (in particular, the nondominant temporal lobe) and mesial temporal structures as critical at the front end of social cognitive processes. The frontal lobes, in turn, function to interpret and to modulate these processes via top-down control. Damage to frontal regions is associated with specific derangements in social behavior. Chronic focal-onset epilepsy and its surgical treatment commonly affect these neuroanatomic regions and might therefore impact social function. Postoperative social function helps determine quality of life for both patients and families. There is some evidence that resective seizure surgery affects social cognition, but there are significant weaknesses in our current knowledge that can be overcome with comprehensive longitudinal research. PMID- 16253568 TI - Effects of antiepileptic drugs on attention as assessed by a five-choice serial reaction time task in rats. AB - Patients with epilepsy can have impaired cognitive abilities. Antiepileptic drugs (AEDs) may contribute to the cognitive deficits observed in patients with epilepsy, and have been shown to induce cognitive impairments in healthy individuals. However, there are few systematic data on the effects of AEDs on specific cognitive domains. We have previously evaluated a number of AEDs with respect to their effects on working memory. The purpose of the present study was to evaluate the effects of AEDs on attention as measured by five-choice serial reaction time behavior in nonepileptic rats. The GABA-related AEDs triazolam, phenobarbital, and chlordiazepoxide significantly disrupted performance by increasing errors of omission, whereas tiagabine, valproate, and gabapentin did not. The sodium channel blocker carbamazepine increased errors of omission at relatively high doses, whereas the sodium channel blockers phenytoin, topiramate, and lamotrigine were without significant effect. Levetiracetam had no effect on attention. The disruptions produced by triazolam, phenobarbital, chlordiazepoxide, and carbamazepine were similar in magnitude to the effects of the muscarinic cholinergic receptor antagonist scopolamine. The present results indicate that AEDs can disrupt attention, but there are differences among AEDs in the magnitude of the disruption in nonepileptic rats, with drugs that enhance GABA receptor function producing the most consistent disruption of attention. PMID- 16253569 TI - Profiling biochemical and hemodynamic markers using chronically instrumented, conscious and unrestrained rats undergoing severe, acute controlled hemorrhagic hypovolemic shock as an integrated in-vivo model system to assess new blood substitutes. AB - The aim of the present study was to assess several biochemical and physiological endpoint parameters alongside controlled hemorrhagic and recovery phases of chronically instrumented, conscious and unrestrained healthy rats. Male Sprague Dawley rats (12-14 weeks; 430+/-20 g; n=22-18) were instrumented with a saline perfused femoral arterial catheter and placed individually in a metabolic cage for up to 20 days, allowing instant assessments of the hemodynamic profile and blood and urine sampling for hematological profile and biochemical measurements to assess hepatic, renal and metabolic functions. In addition, body weight, food and water intake, and diuresis were monitored daily. After a 7-day stabilization period, the rats underwent severe and acute hemorrhagic shock (HS) (removal of 50% of total circulating blood volume), kept in hypovolemic shock for an ischemic period of 50 min and then resuscitated over 10 min. Gr. 1 was re-infused with autologous shed blood (AB; n=10) whereas Gr. 2 was infused 1:1 with a solution of sterile saline-albumin (SA; 7% w/v) (n=8-12). Ischemic rats recovered much more rapidly following AB re-infusion than those receiving SA. Normal hemodynamic and biochemical profiles were re-established after 24 h. Depressed blood pressure lasted 4-5 days in SA rats. The hematological profile in the SA resuscitated rats was even more drastically affected. Circulating plasma concentrations of hemoglobin (-40%), hematocrit (-50%), RBC (-40%) and platelets (-41%) counts were still severely decreased 24 h after the acute ischemic event whereas WBC counts increased 2.2-fold by day 4. It took 5-9 days for these profiles to normalize after ischemia-reperfusion with SA. Diuresis increased in both groups (by 45+/-7% on day 1) but presented distinct electrolytic profiles. Hepatic and renal functions were normal in AB rats whereas altered in SA rats. The present set of experiments enabled us to validate a model of HS in conscious rats and the use of an integrated in vivo platform as a valuable tool to characterize HS-induced stress and to test new classes of blood substitutes in real time, post-event, over days. PMID- 16253570 TI - Voltammetry of the interaction of metronidazole with DNA and its analytical applications. AB - Voltammetric methods were used to probe the interaction of antimicrobial drug metronidazole (MTZ) with calf thymus DNA. Binding constants (K) and binding site sizes (s) were determined from the voltammetric data, i.e., shifts in potential and changes in limiting current with the addition of DNA. MTZ showed appreciable electrostatic binding to DNA in solution with K=2.2(+/- 1.3) x 10(4) M(-1) and s=0.34 bp. One reduction peak of MTZ at the bare glassy carbon electrode (GCE) split into two peaks at the DNA modified GCE (DNA/GCE). These changes in the cyclic voltammogram can only be due to the interaction of MTZ with the surface confined DNA. In addition, the peak current of MTZ at the DNA/GCE was nearly 8 fold of the response at the bare GCE. The low detection limit of 2.0 x 10(-8) M made the DNA/GCE a promising biosensor for MTZ determination. And this method was successfully applied with high precision and accuracy compared with spectroscopic methods (relative error < 6%) for estimation of the total MTZ drug content in pharmaceutical dosage forms. PMID- 16253571 TI - Detection of oxaliplatin-induced DNA crosslinks in vitro and in cancer patients using the alkaline comet assay. AB - Oxaliplatin is frequently used in the therapy of cancer. In DNA, oxaliplatin induces, like cisplatin, the formation of crosslinks, which are commonly accepted as being responsible for the cytotoxicity of platinum agents. The detection of oxaliplatin-induced DNA crosslink formation and repair could be a good measure of assessing how a patient is responding to the agent. In this study, we used a validated modification of the alkaline comet assay for detecting the presence of these crosslinks in vitro and in cancer patients. The H460 tumour cell line was treated in vitro with a range of oxaliplatin and cisplatin doses, and the subsequent crosslink formation and repair compared between the two agents. In addition, lymphocytes from cancer patients undergoing oxaliplatin-based chemotherapy were assayed for the formation and repair of oxaliplatin-induced crosslinks. A dose-response was observed in the in vitro samples, with cisplatin producing more crosslinks than oxaliplatin at equimolar concentrations and lesions induced by both agents showing different repair efficiencies. Furthermore, evidence of crosslink formation and repair was observed in the peripheral blood lymphocytes of all cancer patients studied, along with the detection of interindividual variability in crosslink formation and repair efficiencies. To the best of our knowledge, this is the first time that oxaliplatin DNA crosslinks have been detected either in vitro or in patient samples using the alkaline comet assay. Due to its sensitivity, rapidity, small cell sample and low cost, the alkaline comet assay is a good method for the detection of oxaliplatin-induced crosslinks and their subsequent repair and, in future clinical studies, could prove to be a valuable tool in assessing/predicting a patient's response to chemotherapy. PMID- 16253572 TI - To treat or not to treat with antifungal agents? PMID- 16253573 TI - National scientific associations should have a key role in adapting and implementing standard of care guidelines in European countries. PMID- 16253574 TI - Receptor-ligand binding assays: technologies and applications. AB - Receptor-ligand interactions play a crucial role in biological systems and their measurement forms an important part of modern pharmaceutical development. Numerous assay formats are available that can be used to screen and quantify receptor ligands. In this review, we give an overview over both radioactive and non-radioactive assay technologies with emphasis on the latter. While radioreceptor assays are fast, easy to use and reproducible, their major disadvantage is that they are hazardous to human health, produce radioactive waste, require special laboratory conditions and are thus rather expensive on a large scale. This has led to the development of non-radioactive assays based on optical methods like fluorescence polarization, fluorescence resonance energy transfer or surface plasmon resonance. In light of their application in high throughput screening environments, there has been an emphasis on so called "mix and-measure" assays that do not require separation of bound from free ligand. The advent of recombinant production of receptors has contributed to the increased availability of specific assays and some aspects of the expression of recombinant receptors will be reviewed. Applications of receptor-ligand binding assays described in this review will relate to screening and the quantification of pharmaceuticals in biological matrices. PMID- 16253575 TI - High-performance liquid chromatographic determination of vertilmicin in rat plasma using sensitive fluorometric derivatization. AB - A sensitive and reliable high-performance liquid chromatographic method was developed for the determination of vertilmicin in rat plasma. Derivatization with 9-fluorenylmethyl chloroformate (FMOC-Cl) followed by C(18) reversed-phase chromatography allowed the fluorimetric detection of vertilmicin. Optimal conditions for the derivatization of vertilmicin are described. The limit of quantification was 0.02 mg/L. The pharmacokinetics of vertilmicin was studied in 24 rats following intramuscular injection (i.m.) of different doses (4, 8, 16, 32 mg/kg of body weight). The pharmacokinetic parameter values were estimated by use of 3P97 program. In this study, we assessed the dose proportionality of vertilmicin after single intramuscular injection doses and obtained new information on the pharmacokinetics of the compound. PMID- 16253577 TI - Quantitative scanning acoustic microscopy compared to microradiography for assessment of new bone formation. AB - Recently, it has been shown that quantitative scanning acoustic microscopy (SAM) is a powerful tool to image the acoustic impedance of even inhomogeneous materials like bone. Therefore, the aim of our study was to compare SAM to conventional microradiography with respect to histomorphometrical assessment of undecalcified sections of newly formed bone. Forty specimens were harvested 12 weeks after implantation of either autogenous cancellous bone graft or 5.0 mg of Osteogenic Protein-1 (BMP-7) in a critical-sized defect model in sheep. Undecalcified transverse bone sections of 500 microm thickness were investigated with conventional microradiography and SAM. Linear regression analysis was carried out to compare the measurements of the area of new bone formation within the defect sites. Both methods allowed for good discrimination between newly formed bone and cortical bone at the edges of the former defect. Images obtained with SAM revealed a better resolution and sharpness compared to that of microradiographs since SAM imaging unlike microradiography does not depend on the thickness of bone sections. The results of quantitative histomorphometric analysis obtained by both methods showed no significant differences, and it was possible to predict 90% of the variability of each method (coefficient of determination r2 = 0.90; P < 0.0001). In conclusion, SAM offers comparable quantitative histomorphometric information with a better spatial resolution than conventional microradiography. Thus, SAM is a promising new micro-visualizing technique for basic bone research. PMID- 16253576 TI - Validation and implementation of a liquid chromatography/tandem mass spectrometry assay to quantitate dimethyl benzoylphenylurea (BPU) and its five metabolites in human plasma and urine for clinical pharmacology studies. AB - A method has been developed for the quantitation of N-[4-(5-bromo-2 pyrimidinyloxy)-3-methylphenyl]-N'-(2-dimethylamino-benzoyl)urea (BPU) and its metabolites in human plasma and urine. BPU and metabolites were separated on a C18 column with acetonitrile-water mobile phase containing 0.1% formic acid using isocratic flow for 5 min. The analytes were monitored by tandem mass spectrometry. Calibration curves were generated over the range of 2.5-500 ng/mL for BPU, mmBPU, and aminoBPU in plasma; and 0.1-20, 0.1-20, 0.5-100, 10-2000, 1 200, and 3-600 ng/mL for BPU, mmBPU, aminoBPU, G280, G308, and G322 in urine, respectively. The method has been successfully applied to study the pharmacokinetics of BPU. PMID- 16253578 TI - Effect of depression on five-year mortality after an acute coronary syndrome. AB - Previous research has established a relation between depression at the time of cardiac hospitalization and patient mortality. The objective of this study was to examine the role of depressive history and symptomatology during hospitalization on 5-year all-cause mortality after admission for an acute coronary syndrome. We recruited 750 patients who had unstable angina pectoris and myocardial infarction from 12 coronary care units between 1997 and 1999. Measurements included sociodemographic and clinic data and the Beck Depression Inventory (BDI). Data were linked to an administrative database to determine 5-year all-cause mortality. Survival data were adjusted using a Cox's proportional hazards model. One hundred seventy-four participants (23.2%) self-reported a history of depressed mood for >2 weeks, 235 (31.3%) had elevated BDI scores at index hospitalization, with 105 (14.0%) reporting persistent depressive symptomatology. One hundred fifteen participants (15.3%) died by 5 years after hospitalization. After adjusting for prognostic indicators, such as cardiac disease severity, medical history, and smoking, depressive symptomatology during hospitalization was significantly predictive of mortality, but depressive history was not. Hazard ratios associated with BDI scores <10 versus those > or =10 at hospitalization ranged from 1.90 (95% confidence interval 1.12 to 3.24) at 2 years to 1.53 (95% confidence interval 1.04 to 2.24) at 5 years. In conclusion, the significance of depressive symptomatology at the time of, but not before, hospitalization underlines the need for early identification of increased distress and renews calls to identify treatments that not only improve quality of life but also decrease the risk of mortality. PMID- 16253579 TI - Usefulness of left ventricular diastolic wall motion abnormality as an early predictor of left ventricular dilation after a first acute myocardial infarction. AB - To determine the relation between regional diastolic wall motion abnormality and left ventricular remodeling after acute myocardial infarction (AMI), Doppler echocardiography and color kinesis with assessment of global and regional systolic and diastolic functions were performed in 84 patients who developed AMI within 24 hours of admission. In a multivariate logistic regression analysis, the percentage of left ventricular myocardial segments with diastolic wall motion abnormality (p = 0.008), absence of myocardial viability (p = 0.01), and overall diastolic function (p = 0.001) were predictors of remodeling after AMI. PMID- 16253580 TI - A multistate comparison of patient characteristics, outcomes, and treatment practices in acute myocardial infarction. AB - The primary purpose of this study was to examine variations in patient characteristics, outcomes, and treatment practices in acute myocardial infarction (AMI) across 11 states. Data from 11 states were extracted from the Healthcare Cost and Utilization Project State Inpatient Dataset. Patients who had a primary diagnosis of AMI (International Classification of Diseases, Ninth Revision, Clinical Modification, code 410.x1) from 11 states were extracted from the Healthcare Cost and Utilization Project 1999 dataset. Bivariate comparisons were conducted to examine the characteristics, treatment practices, and outcomes of patients who had AMI. Multivariate regression models were used to examine the association between geographic location (and other factors) and the likelihood of in-hospital mortality, undergoing coronary artery bypass grafting (CABG), or percutaneous coronary interventions (PCIs). Results revealed considerable variations across states in practice patterns and treatment outcomes. New York had the highest average length of stay (8.2 days, p <0.01), rate of patients who had AMI being transferred (20.7%, p <0.01), and in-hospital case fatality rate (10.7%, p <0.01) and the lowest rate of alive discharges being routine (65.6%, p <0.01). PCI was performed 2 times as often as CABG for patients who had AMI (23.9% vs 11.3%, p <0.01), with patients who underwent CABG being transferred more often. Multivariate analyses showed that state of residence, age, female gender, transfer status, and number of co-morbidities were predictors of in hospital mortality and the likelihood of undergoing CABG or PCI. In conclusion, large differences in practice patterns and treatment outcomes exist across states. PMID- 16253581 TI - N-terminal pro-brain natriuretic peptide and tombstoning ST-segment elevation in patients with anterior wall acute myocardial infarction. AB - Tombstoning ST-segment elevation myocardial infarction (STEMI) has been associated with a poor prognosis ever since Wimalaratna's first description of this clinical entity, and the reasons for this are not fully understood. We studied 87 consecutive patients who had anterior STEMI (January 2004 to September 2004) to compare the form of STEMI, coronary anatomy, and initial N-terminus pro brain natriuretic peptide (NT-pro-BNP) level. Patients who had tombstoning STEMI had higher levels of NT-pro-BNP despite significant differences in cardiac enzyme levels or extent of coronary disease. This finding suggests that, in addition to changing the shape of repolarization, decreased microcirculation plays a role in the development of increased wall tension. Increased wall tension in turn is the probable cause of higher NT-pro-BNP levels and increased mortality. PMID- 16253582 TI - Impact of bleeding severity on clinical outcomes among patients with acute coronary syndromes. AB - Bleeding is a complication of current therapies for acute coronary syndrome (ACS). No studies have examined the effect of bleeding events on clinical outcomes. We analyzed pooled data from 4 multicenter, randomized clinical trials of patients who had ACS (n = 26,452) to determine an association between bleeding severity as measured by the GUSTO scale and 30-day and 6-month mortality rates using Cox proportional hazards modeling that incorporated bleeding as a time dependent covariate. The analysis was repeated to examine procedure- and non procedure-related bleeding and after censoring at the time of coronary artery bypass grafting. Of all the patients included, 27.6% had > or =1 bleeding episode. Patients who bled were older and sicker at presentation than were those who did not bleed. Unadjusted rates of 30-day and 6-month mortality increased as bleeding severity increased. There were stepwise increases in the adjusted hazards of 30-day mortality (mild bleeding, hazard ratio [HR] 1.6, 95% confidence interval [CI] 1.3 to 1.9; moderate bleeding, HR 2.7, 95% CI l 2.3 to 3.4; severe bleeding, HR 10.6, 95% CI 8.3 to 13.6) and 6-month mortality (mild bleeding, HR 1.4, 95% CI 1.2 to 1.6; moderate bleeding, HR 2.1, 95% CI 1.8 to 2.4; severe bleeding, HR 7.5, 95% CI 6.1 to 9.3) as bleeding severity increased. Results were consistent after censoring for coronary artery bypass grafting and for procedure- and non-procedure-related bleeds. In conclusion, the GUSTO bleeding classification identifies patients who are at risk for short- and long-term adverse events. Therapies that minimize bleeding risk and maintain an anticoagulant effect may improve outcomes among patients who have ACS. PMID- 16253584 TI - Frequency of abdominal aortic aneurysm in patients >60 years of age with coronary artery disease. AB - Using B-mode ultrasound, we studied the prevalence of abdominal aortic aneurysm (AAA; diameter > or =3 cm) and its predictive risk factors in 109 consecutive patients who were >60 years of age and had coronary artery disease (CAD). A group of 60 age-matched patients who did not have CAD served as controls. The prevalence of AAA was higher in the CAD group than in the control group (14%, 16 of 109, vs 3%, 2 of 60, p <0.05). By multivariate analysis, only smoking was strongly associated with AAA (odds ratio 4.86, 95% confidence interval 1.55 to 15.25). In contrast, presence of diabetes mellitus was negatively associated with AAA in univariate analysis (odds ratio 0.11, 95% confidence interval 0.01 to 0.83) and a strong trend of inverse association remained in multivariate analysis (odds ratio 0.12, 95% confidence interval 0.01 to 1.03). Thus, systematic screening can detect AAA in 1 of 7 patients who are >60 years of age and have CAD. AAA shares some, but not all, risk factors of atherosclerosis. PMID- 16253583 TI - Intensity of lipid lowering with statins and brachial artery vascular endothelium reactivity after acute coronary syndromes (from the BRAVER trial). AB - The time course and differential effects of statin regimens on endothelial function after acute coronary syndromes (ACSs) are unknown and could contribute to the superiority of a more intense strategy. A subset of subjects who were enrolled in the PROVE IT-TIMI 22 trial (n = 50) underwent evaluation of vascular reactivity by high-resolution brachial ultrasound. Endothelium-dependent flow mediated dilation (FMD) and endothelium-independent sublingual nitroglycerin mediated dilation (NMD) were measured at baseline and at 48 hours, 1 month, and 4 months after the initiation of 40 mg of pravastatin (n = 26) or 80 mg of atorvastatin (n = 24). After 4 months, low-density lipoprotein cholesterol was decreased by 32% in the atorvastatin group but was not different from baseline after ACS in the pravastatin group. C-reactive protein decreased similarly in the 2 groups. Brachial artery diameters at rest were similar in the 2 groups and at each time point of the trial. FMD and NMD increased significantly after 4 months by 27% and 24%, respectively (p <0.05), with no difference between groups. There was no correlation between the change in FMD and the change in lipids or C reactive protein. In subjects who had received previous statin therapy (n = 15), there was no significant variation in FMD (p = 0.140) and NMD (p = 0.129). In conclusion, initiation of statin therapy soon after ACS is associated with improvements in endothelium-dependent and independent vascular reactivities after 4 months. PMID- 16253585 TI - Gender differences in carotid intima-media thickness in patients with suspected coronary artery disease. AB - Selecting patients who have suspected coronary artery disease (CAD) for coronary angiography remains difficult in some subgroups (e.g., women have often atypical symptoms and false-positive findings on noninvasive tests). This study evaluated gender differences and the clinical value of carotid intima-media thickness (IMT) in predicting CAD. We evaluated 558 patients who had symptoms and underwent coronary angiography: 91 women (61.2 +/- 9.5 years of age) had CAD (> or =1 lesion with a decrease >50% in luminal diameter of a coronary artery), 29 women (57.9 +/- 7.0 years of age) did not have CAD, 372 men (58.9 +/- 9.2 years of age) had CAD, and 66 men (54.6 +/- 8.7 years of age) did not have CAD. Maximal IMT was assessed bilaterally at the common carotid, bulb, and internal carotid arteries and expressed as mean IMT for each patient. Among patients who did not have CAD, women had lower mean IMT values than men (0.93 +/- 0.15 vs 1.05 +/- 0.19, p < 0.001). This gender difference was not seen in patients who had CAD (1.3 +/- 0.31 vs 1.31 +/- 0.31, p = 0.92). Among women and men, those with CAD had larger IMT values than those without CAD. Multivariable regression analysis showed that age, CAD, hypertension, smoking, and diabetes had the strongest effect on IMT values in women and men. Receiver-operator characteristic analysis showed that women had a significantly lower IMT threshold for likelihood of CAD (p < 0.001) and that a mean IMT of 1.069 mm was highly predictive of concomitant CAD (sensitivity 79%, specificity 90%, positive predictive value 96%); for men, the mean IMT threshold was 1.153 mm (sensitivity 66%, specificity 74%, positive predictive value 93%). In conclusion, carotid IMT assessment may be a valuable tool in selecting patients for coronary angiography to predict the likelihood of CAD. This particularly concerns women in whom sensitivity and specificity of mean IMT are high. PMID- 16253586 TI - Gender-based differences in brachial artery flow-mediated vasodilation as an indicator of significant coronary artery disease. AB - Ultrasound of the brachial artery is widely used to assess endothelial function, but whether brachial artery flow-mediated vasodilation (FMD) differs between women and men who have coronary artery disease (CAD) has not been examined. To investigate gender-based differences in brachial artery FMD as an indicator of significant CAD, FMD was measured in women and men outpatients who had CAD (coronary stenosis >50%, n = 64) and those who did not have significant CAD (n = 145). FMD in women who had CAD (n = 33, 9.1 +/- 0.8%) was higher than that in similarly aged men who had CAD (n = 31, 6.4 +/- 0.5%; p = 0.008). The FMD cutpoint that maximized sensitivity with least effect on specificity for screening CAD was 15% (91% sensitivity, 25% specificity) in women but 10% (90% sensitivity, 43% specificity) in men. If the cutpoint as defined in men were used to evaluate women, brachial artery ultrasound would fail to diagnose 42% of women who do not have significant CAD; thus, a higher FMD cutpoint is required to optimize the sensitivity of FMD for identifying women who have significant CAD compared with similarly aged men. In studies using FMD to evaluate cardiovascular risk, different standards should be applied for women and men. PMID- 16253587 TI - Outcome in patients transferred for percutaneous coronary intervention (a national registry of myocardial infarction 2/3/4 analysis). AB - Hospital transfer is associated with delays in performance of primary percutaneous coronary intervention (PCI) for ST-elevation myocardial infarction. Given the limited number of interventional centers in the United States, the transfer of patients is widely practiced. We compared the outcome of patients who were transferred for primary PCI and determined whether prolonged door-to-balloon times were associated with increased mortality. Patients who presented within 12 hours with ST-segment elevation myocardial infarction and underwent transfer for primary PCI were identified from the National Registry of Myocardial Infarction database (NRMI databases 2, 3, and 4, n = 7,133). The short-term outcome of those who received early (< or =2 hours) was compared with that of those who received delayed primary PCI (>2 hours) using multivariate logistic regression analyses and propensity score methods. The door-to-balloon time for the early PCI group compared with the delayed PCI group was 99 +/- 16 versus 264 +/- 178 minutes, respectively (p <0.0001). The early PCI group had less recurrent ischemia and angina (5.8% vs 10.1%, p <0.001), less cardiogenic shock (5.1% vs 8.9%, p <0.001), and shorter length of hospital stay (4.4 +/- 3.5 vs 5.4 +/- 4.7 days, p <0.001). In-hospital mortality was lower for the early PCI group than for the delayed PCI group (2.7% vs 6.2%, p < 0.001; entire cohort 5.7%). Comparison of patients matched on propensity score (n = 993) showed that mortality was lower in the early than in the delayed PCI group (2.6% vs 4.6%, p = 0.014, c-statistic 0.67). In conclusion, <4% of patients who received PCI were treated within the recommended guideline of < 120 minutes by the American College of Cardiology/American Heart Association. Door-to-balloon times <2 hours in patients who undergo transfer for PCI is associated with a significant decrease in short term mortality, which suggests that efforts must be made to decrease transfer delays. PMID- 16253588 TI - Relation of terminal QRS distortion to left ventricular functional recovery and remodeling in acute myocardial infarction treated with primary angioplasty. AB - The association between admission electrocardiogram and 6-month change in left ventricular function and volume was assessed in 200 patients who had acute myocardial infarction that was treated with primary percutaneous coronary intervention. Logistic regression analysis indicated peak creatine phosphokinase MB, number of Q-wave leads, QRS interval distortion, wall motion score index, and angiographic Thrombolysis In Myocardial Infarction flow grade as predictors of no functional recovery and QRS interval distortion and Thrombolysis In Myocardial Infarction flow grade as predictors of left ventricular remodeling. PMID- 16253589 TI - Impact of intravascular ultrasound lesion characteristics on neointimal hyperplasia following sirolimus-eluting stent implantation. AB - The effect of lesion characteristics on neointimal hyperplasia after sirolimus eluting stent implantation was examined in 45 patients who underwent successful preinterventional intravascular ultrasound. There were no differences in neointimal hyperplasia between the moderate/severe calcified lesion group (calcium arc >120 degrees ) and the non/mild calcified lesion group or between the positive vessel remodeling group (external elastic membrane area at the minimal lumen area site larger than that at the proximal reference site) and negative vessel remodeling group. No correlation between preinterventional plaque burden and neointimal hyperplasia was found. In patients who have coronary artery disease, sirolimus-eluting stents continue to demonstrate striking suppression of neointimal proliferation, irrespective of lesion characteristics previously associated with greater restenotic risk. PMID- 16253591 TI - Neurological complications following percutaneous coronary interventions (a report from the 2000-2001 New York State Angioplasty Registry). AB - Stroke is 1 of the most devastating complications associated with percutaneous coronary intervention. The present study used the combined 2000 to 2001 New York State Angioplasty Registry to compare the clinical characteristics and in hospital outcomes of patients with and without stroke after percutaneous coronary intervention. Of the 76,903 patients who underwent angioplasty, 140 (0.18%) experienced stroke. Multivariate regression analysis revealed age, glycoprotein IIb/IIIa inhibitor use, acute myocardial infarction or congestive heart failure on admission, history of carotid disease, chronic renal disease, and placement of an intra-aortic balloon pump as independent predictors for stroke complicating percutaneous coronary intervention. PMID- 16253590 TI - Impact of coronary culprit lesion calcium in patients undergoing paclitaxel eluting stent implantation (a TAXUS-IV sub study). AB - Randomized clinical trials have shown that paclitaxel-eluting stents significantly reduce restenosis after percutaneous coronary intervention. The impact of lesion calcification on the efficacy of paclitaxel-eluting stents is unknown. In the TAXUS-IV trial, 1,314 patients who underwent percutaneous coronary intervention were randomly assigned to a bare-metal or paclitaxel eluting stent. By core laboratory analysis, 247 lesions (19%) were moderately or severely calcified. At the 9-month angiographic follow-up examination, the paclitaxel-eluting stent had significantly reduced the amount of late loss compared with the control stent (0.26 +/- 0.56 vs 0.51 +/- 0.48 mm, p = 0.015) within the analysis segment in the calcific lesions. The analysis segment restenosis rate was similar in patients with calcified and noncalcified lesions after paclitaxel-eluting stent implantation (7.5% vs 8.0%, respectively; p = 1.0). The rate of ischemia-driven target lesion revascularization (TLR) at 1 year was reduced by 56% in patients with calcified lesions (11.9% vs 5.1%, p = 0.09) and by 75% in noncalcified lesions (15.7% vs 4.3%, p <0.0001). By interaction testing, the efficacy of the paclitaxel-eluting stent in reducing TLR at 1 year was similar in the calcified and noncalcified lesions (p = 0.30). Moreover, by multivariate analysis, implantation of the paclitaxel-eluting stent was a powerful independent predictor of freedom from TLR, with similar hazard ratios for efficacy in calcified and noncalcified lesions (0.30 and 0.26, respectively). In conclusion, implantation of paclitaxel-eluting stents in patients with de novo coronary lesions significantly reduced restenosis in patients with and without calcified lesions. PMID- 16253592 TI - Predictors of edge stenosis following sirolimus-eluting stent deployment (a quantitative intravascular ultrasound analysis from the SIRIUS trial). AB - To study the interaction of the sirolimus-eluting stent and vessel margins, we analyzed the intravascular ultrasound parameters in 317 edges of 167 stents having 18 edge stenoses at 8 months of follow-up from the SIRIUS trial. Of the baseline parameters, a larger reference percentage of plaque area and a larger edge stent area/reference minimum lumen area were associated with edge stenosis in the sirolimus-eluting stent cohort compared with the incidence of edge stenosis in the bare metal stent cohort. Thus, full lesion coverage and matching the stented segment properly to the adjacent segment using intravascular ultrasound guidance may improve sirolimus-eluting stent implantation efficacy further. PMID- 16253593 TI - Relation of early saphenous vein graft failure to outcomes following coronary artery bypass surgery. AB - Up to 20% of saphenous vein grafts (SVGs) fail within 2 years of coronary artery bypass grafting (CABG). The long-term effects of early SVG failure on major clinical events remain undefined in contemporary patient populations. We sought to examine the relation between early SVG failure and long-term outcomes after CABG. Using the Duke Cardiovascular Databank, we examined baseline clinical and angiographic characteristics and clinical outcomes among patients who underwent catheterization 1 to 18 months after their first CABG from 1986 to 2004. Patients were classified on the basis of their worst SVG stenosis as having no (<25%), noncritical (25% to 74%), critical (75% to 99%), or occlusive (100%) SVG disease. Our primary outcome measure was the composite of death, myocardial infarction, or repeat revascularization after catheterization. Of 1,243 patients included in the analysis, 27.9% had no, 11.9% had noncritical, 20.8% had critical, and 39.3% had occlusive SVG disease. At 10 years, the corresponding adjusted composite event rates were 41.2%, 56.2%, 81.2%, and 67.1%, respectively (p <0.0001). Most events occurred immediately after catheterization in patients with critical and occlusive SVG disease and were primarily repeat revascularization. On multivariate analysis, critical, nonocclusive SVG disease was the strongest predictor of the composite outcome (hazard ratio 2.36, 95% confidence interval 2.00 to 2.79, p <0.0001). In conclusion, in contemporary clinical practice, early SVG failure is associated with worse long-term outcomes after CABG. PMID- 16253594 TI - Therapeutic lifestyle changes and drug treatment for high blood cholesterol in China and application of the Adult Treatment Panel III guidelines. AB - The prevalence of elevated blood cholesterol in China has increased during the past several decades. We estimated the percentage of the Chinese population for whom therapeutic lifestyle changes and drug therapy to lower blood cholesterol should be considered by applying the United States' National Cholesterol Education Panel's Adult Treatment Panel III guidelines to a nationally representative sample of the Chinese population from the International Collaborative Study of Cardiovascular Disease in Asia. Serum samples were collected for 14,919 Chinese adults, 35 to 74 years old, in 2000 and 2001, after an overnight fast of > or =8 hours and their low-density lipoprotein (LDL) cholesterol level was calculated using the Freidewald equation. Using the Adult Treatment Panel III guidelines, 85.9 million Chinese adults (18.2%) should initiate therapeutic lifestyle changes to lower their LDL cholesterol and 35.0 million (7.4%) should be considered for lifestyle changes and lipid-lowering drug therapy. Of those for whom drug therapy should be considered, 4.7 million (13.4%) reported having been told they had "high cholesterol" by a healthcare provider and 1.6 million (33.7% of those aware of their high cholesterol) were receiving lipid-lowering medication-leaving 33.4 million Chinese adults with untreated elevated LDL cholesterol (95.5% of those with elevated LDL cholesterol). A 10% population-wide reduction in LDL cholesterol would reduce the number of Chinese adults who should be considered for drug therapy by 45% to 19.3 million (4.1% of adults). In conclusion, most adults in China with an elevated LDL cholesterol remain untreated. PMID- 16253595 TI - Gemfibrozil reduces small low-density lipoprotein more in normolipemic subjects classified as low-density lipoprotein pattern B compared with pattern A. AB - We tested the hypothesis that gemfibrozil has a differential effect on low density lipoprotein (LDL) and high-density lipoprotein (HDL) subclass distributions and postprandial lipemia that is different in subjects classified as having LDL subclass pattern A or LDL pattern B who do not have a classic lipid disorder. Forty-three normolipemic subjects were randomized to gemfibrozil (1,200 mg/day) or placebo for 12 weeks. Lipids and lipoproteins were determined by enzymatic methods. The mass concentrations of lipoproteins in plasma were determined by analytic ultracentrifugation and included the S(f) intervals: 20 to 400 (very LDL), 12 to 20 (intermediate-density lipoprotein), 0 to 12 (LDL), and HDL(2) mass (F(1.20) 3.5 to 9.0) and HDL(3) mass (F(1.20) 0 to 3.5). Postprandial measurements of triglycerides and lipoprotein(a) were taken after the patients consumed a 500 kcal/M(2) test meal. Treatment with gemfibrozil, compared with placebo, significantly reduced fasting plasma triglycerides (difference from placebo +/- SE; -50.2 +/- 20.6 mg/dl, p = 0.02), total cholesterol (-16.4 +/- 7.5 mg/dl, p = 0.04), apolipoprotein B (-16.1 +/- 5.5 mg/dl, p = 0.006), very LDL mass of S(f) 20 to 400 (-50.8 +/- 24.1 mg/dl, p = 0.02), S(f) 20 to 60 (-17.5 +/- 8.5 mg/dl, p = 0.05), S(f) 60 to 100 (-16.2 +/- 8.1 mg/dl, p = 0.05), and increased peak S(F) (0.48 +/- 0.27 Svedberg, p = 0.08). Gemfibrozil reduced the postprandial triglyceride level significantly at 3 (p = 0.04) and 4 (p = 0.05) hours after the test meal. A significantly different subclass response to gemfibrozil was observed in those with LDL pattern A versus B. Those with LDL pattern B had a significantly greater reduction in the small LDL mass S(f) 0 to 7 (p = 0.04), specifically regions S(f) 0 to 3 (p = 0.009) and S(f) 3 to 5 (p = 0.009). In conclusion, normolipemic subjects with either predominantly dense or buoyant LDL respond differently to gemfibrozil as determined by the changes in LDL subclass distribution. Thus, treatment with gemfibrozil may have additional antiatherogenic effects in those with LDL pattern B by decreasing small dense LDL that is not apparent in those with pattern A. PMID- 16253596 TI - Relation of plasma glucose and endothelial function in a population-based multiethnic sample of subjects without diabetes mellitus. AB - To determine whether endothelial dysfunction precedes the clinical diagnosis of diabetes mellitus, we investigated the relation of endothelial flow-mediated dilation (FMD) with fasting plasma glucose among a multiethnic population-based cohort of 579 nondiabetic subjects without previous myocardial infarction or stroke enrolled in the Northern Manhattan Study (age 66 +/- 9 years; 41% men, 16% white, 15% black, and 68% Hispanic). Impaired fasting glucose or prediabetic status, defined as a fasting glucose level of 100 to 125 mg/dl, was present in 95 subjects (16%). Endothelial function was determined using FMD during reactive hyperemia. Multiple linear regression analyses were used to assess the relation between plasma glucose and endothelial function after adjustment for potential confounders. FMD was significantly lower (4.9 +/- 3.8% vs 6.1 +/- 3.7%, p = 0.003) in those with impaired fasting glucose than in subjects with normal fasting glucose. Prediabetic status was significantly associated with impaired FMD (odds ratio 1.9, 95% confidence interval 1.1 to 3.1, p = 0.02). After adjustment for age, gender, body mass index, and hypertensive status, a higher fasting glucose was significantly associated with a lower FMD (beta = -0.024 +/- 0.012, p = 0.04) in a continuous linear relation. Thus, for each 10-mg/dl increase in plasma glucose, a 0.24% decrease occurred in FMD. Impaired FMD was present among prediabetics. An elevated fasting plasma glucose level is associated with impaired endothelial function among nondiabetics. These results further support the role of hyperglycemia in the pathogenesis of vascular dysfunction at different stages of diabetes development and the role of impaired fasting glucose as a risk factor for macrovascular disease. PMID- 16253597 TI - Experience with statin use in patients with chronic hepatitis C infection. AB - We reviewed the liver enzymes of patients with chronic hepatitis C infection currently taking statin drugs. We found no significant elevation of liver enzymes during statin treatment of the 17 patients reviewed. PMID- 16253598 TI - Relation of the prothrombotic state to increasing age (from the Framingham Offspring Study). AB - A greater life expectancy has led to an increasing proportion of elderly patients. Increasing age is an important risk factor for cardiovascular disease, but the mechanism of risk is not well understood. Because thrombosis plays a key role in plaque development and the onset of acute coronary syndromes, the age related increase in cardiovascular risk may be a result of a prothrombotic imbalance. The study aim was to examine the relation between age and thrombotic potential in the Framingham Offspring Cohort. Hemostatic factors previously associated with cardiovascular risk were measured in 3,230 patients (55% women) without evidence of cardiovascular disease who were participating in cycle 5 of the Framingham Offspring Study. The subjects were divided by age into decades. Advancing age was associated with a significant increase in fibrinogen and von Willebrand factor levels and measures of impaired fibrinolytic potential (plasminogen activator inhibitor and tissue plasminogen activator antigens). For men, the mean fibrinogen levels were 21% higher in those > or =70 years versus those aged <40 years (326 vs 268 mg/dl, p <0.001 for linear trend). The mean fibrinogen levels were 15% higher in older than in younger women (330 vs 286 mg/dl, p <0.001). The significant relations persisted after multivariate adjustment. In conclusion, advancing age is associated with elevated levels of hemostatic factors indicative of a prothrombotic state. Because these factors are also associated with endothelial dysfunction, these findings are consistent with an injurious effect of age on the endothelium. Measures to reduce thrombotic potential may be of particular value in the elderly, because they counter the prothrombotic state that develops with aging. PMID- 16253599 TI - Relation of coronary atherosclerosis determined by electron beam computed tomography and plasma levels of n-terminal pro-brain natriuretic peptide in a multiethnic population-based sample (the Dallas Heart Study). AB - Elevated plasma levels of B-type natriuretic peptide (BNP) and N-terminal pro-BNP (NT-pro-BNP) are seen in the setting of cardiac ischemia and are associated with adverse outcomes in patients with coronary artery disease. The mechanisms leading to natriuretic peptide elevation in patients with coronary artery disease, including the contribution of coronary atherosclerosis itself, have not been fully elucidated. Measurement of NT-pro-BNP, electron beam computed tomography, and cardiac magnetic resonance imaging were performed in 2,445 subjects from the Dallas Heart Study who were free of heart failure and renal insufficiency. Electron beam computed tomography-determined coronary artery calcium scores were categorized as none (<10), mild (> or =10 to <100), moderate (> or =100 to <400), and severe (> or =400). NT-pro-BNP levels increased significantly across increasing coronary artery calcium score categories (p <0.0001 for trend). In multivariate models adjusted for age, gender, race, body mass index, hypertension, history of myocardial infarction, angina, angiotensin-converting enzyme inhibitor use, beta-blocker use, left ventricular (LV) ejection fraction, and LV mass, higher coronary artery calcium scores remained independently associated with higher log NT-pro-BNP levels (p = 0.03). This association persisted in similar models excluding patients with low LV ejection fractions, LV hypertrophy, angina pectoris, and a history of myocardial infarction. In conclusion, these findings support the hypothesis that coronary atherosclerosis may directly influence the activation of the cardiac neurohormonal system. PMID- 16253600 TI - Effect of rosuvastatin on C-reactive protein and renal function in patients with chronic kidney disease. AB - The purpose of this 20-week, open-label, randomized clinical trial was to evaluate the effect of rosuvastatin on fasting serum lipids and lipoproteins, high-sensitivity C-reactive protein (hs-CRP), and the glomerular filtration rate (GFR) in 91 patients with chronic kidney disease. Patients were randomized to rosuvastatin 10 mg/day (n = 48) or to no lipid-lowering treatment (n = 43) for 20 weeks. In contrast to patients not receiving rosuvastatin, patients receiving rosuvastatin tended to derive more favorable improvements from baseline values in low-density lipoprotein cholesterol (-43%, p <0.001, vs 7%, p = NS; p <0.001 for change with rosuvastatin treatment vs change with no antilipemic treatment), hs CRP (-47%, p <0.001, vs 7%, p = NS; p <0.001 for change with rosuvastatin treatment vs change with no antilipemic treatment), and GFR (11%, p <0.05, vs 4%, p = NS; p = NS for change with rosuvastatin treatment vs change with no antilipemic treatment). PMID- 16253601 TI - Prognostic significance of nonsustained ventricular tachycardia during dobutamine stress echocardiography. AB - Nonsustained ventricular tachycardia (NSVT) is a well-recognized side effect during dobutamine stress echocardiography (DSE). This study sought to evaluate the prognostic implications of NSVT during DSE on 1,266 consecutive dobutamine stress echocardiograms performed over 1 year. NSVT, defined as > or =3 consecutive ventricular premature beats, occurred in 65 of 1,266 patients (5.1%). There was no absolute increased risk in all-cause mortality between the NSVT and no NSVT groups (22% vs 17%, p = 0.15) during the 3-year follow-up. Survival curves generated by the Kaplan-Meier method also demonstrated no increased risk in mortality between the NSVT and no NSVT groups (p = 0.43). When only studies with negative results for inducible ischemia were taken into account, survival curves showed no significant difference in all-cause mortality (p = 0.26). Studies with negative results for inducible ischemia were also stratified according to the ejection fraction (EF). Patients without inducible ischemia and mildly reduced to normal EFs (>0.45) did not have significant differences in survival between the NSVT and no NSVT groups over the 3-year follow-up (p = 0.86). However, patients without inducible ischemia and moderately reduced EFs (0.35 to 0.45) who had NSVT during DSE had significantly reduced survival over the follow-up (p = 0.01). PMID- 16253603 TI - Effect of dynamic left ventricular dyssynchrony on dynamic mitral regurgitation in patients with heart failure due to coronary artery disease. AB - In patients with heart failure, exercise-induced increases in mitral regurgitation (MR), which convey a poor prognosis, are related to the dynamic distortion of mitral valve geometry. It was hypothesized that dynamic MR may also be related to intermittent changes in left ventricular synchronicity during exercise. PMID- 16253602 TI - Analysis of left ventricular systolic function using midwall mechanics in patients >60 years of age with hypertensive heart disease and heart failure. AB - Normal ejection fraction (EFs) is often equated with normal systolic function. However, midwall mechanics reveal systolic dysfunction in hypertensive heart disease accompanied by hypertrophic remodeling. Midwall mechanics are unstudied in patients with acute diastolic heart failure (HF). This study analyzed left ventricular (LV) midwall stress-shortening relations in 61 patients aged >60 years with hypertensive heart disease, HF, and normal EF. Sixty-one hypertensive patients (mean age 78 +/- 10 years) who presented with HF, each with an EF >50%, underwent echocardiography. Midwall mechanics were compared with those of 79 controls (mean age 75 +/- 8 years) without structural heart disease. Relative wall thickness (0.63 +/- 0.11 vs 0.46 +/- 0.10 mm) and LV mass (237 +/- 67 vs 177 +/- 57 g) were significantly greater in patients with HF compared with controls. Mean EFs were similar in patients with HF and controls (64 +/- 9% vs 67 +/- 9%). Although mean endocardial fractional shortening (35 +/- 7% vs 37 +/- 7%) was not significantly different, midwall shortening in patients with HF was significantly less compared with controls (16 +/- 2% vs 19 +/- 3%, p <0.05). Eighteen of the 61 patients with HF (30%) had midwall shortening that was <95% confidence intervals of the normal midwall stress-shortening relations. By this criterion, these patients had systolic dysfunction despite normal EF; they had smaller LV chambers (in dimension and volume), greater relative wall thickness, and smaller stroke volumes. In conclusion, almost 1/3 of patients hospitalized with diastolic HF had systolic dysfunction, characterized by abnormal midwall stress-shortening relations. PMID- 16253604 TI - Differences in diagnostic value of four electrocardiographic voltage criteria for hypertrophic cardiomyopathy in a genotyped population. AB - The diagnostic value of various classic electrocardiographic (ECG) voltage criteria for hypertrophic cardiomyopathy (HC) has not been established in a genotyped population. This study aimed to determine the most accurate diagnostic definition of classic ECG voltage criteria for detecting carriers of HC. ECG and echocardiographic findings were analyzed in 161 genotyped subjects (97 genetically affected, 64 unaffected) from 20 families with disease-causing mutations in 4 genes. The diagnostic value of 4 voltage criteria (Cornell, Sokolow-Lyon, Romhilt-Estes, and 12-lead QRS voltage) for detecting carriers of HC was investigated. In all subjects, the Romhilt-Estes (point score > or =4) criterion and 12-lead QRS voltage (> or =240 mm) were most sensitive (37% and 36%, respectively), with high specificity (95% each), resulting in the greatest accuracy (60% and 59%, respectively). Using these criteria, in subjects without echocardiographic evidence of left ventricular hypertrophy, voltage abnormalities were found in 22.6% of carriers and 4.7% of noncarriers (p <0.01). In conclusion, these findings suggest that the Romhilt-Estes and the 12-lead QRS voltage criteria may be the most accurate diagnostic definitions for HC on the basis of molecular genetic diagnoses. Furthermore, this study demonstrated that voltage abnormalities may be found in prehypertrophic carriers. Even when genetic testing becomes widely available, it will be difficult to make genetic diagnoses in all patients with HC because of its genetic heterogeneity. Therefore, understanding the diagnostic value of classic ECG voltage criteria may be important in detecting carriers, including those without left ventricular hypertrophy. PMID- 16253605 TI - Usefulness of magnetic resonance angiography for diagnosis of scimitar syndrome in early infancy. AB - This study evaluated the feasibility and accuracy of the magnetic resonance angiography (MRA) evaluation of scimitar syndrome in early infancy in 4 subjects. MRA imaging was successful, with excellent imaging quality to evaluate the main and lobar pulmonary artery branches, lobar pulmonary veins, scimitar vein, and systemic-pulmonary collateral arteries in all subjects. There was good agreement between findings with MRA, x-ray angiography (n = 3), surgical inspection (n = 3), and autopsy (n = 1). PMID- 16253606 TI - Echocardiographic Doppler assessment of the biophysical properties of the aorta in pediatric patients with the Marfan syndrome. AB - In the Marfan syndrome (MS), aortic root involvement is usually clinically monitored according to the aortic sinus of Valsalva dilation. Using an echocardiographic Doppler method in this cross-sectional study, abnormal biophysical properties of the ascending aorta were found in pediatric patients with MS compared with healthy controls but not consistently in association with the aortic sinus of Valsalva dilation. PMID- 16253607 TI - Prospective evaluation of aggressive medical therapy for atherosclerotic renal artery stenosis, with renal artery stenting reserved for previously injured heart, brain, or kidney. AB - Sixty-six patients with atherosclerotic renal artery stenosis (RAS) and serum creatinine < or =2.0 mg/dl were treated with antihypertensive therapy, a statin, and aspirin. Renal stenting was reserved for patients with injuries to the heart, brain, or kidneys. The primary end point was stenotic kidney glomerular filtration rate (GFR) at 21 months; secondary end points included major adverse clinical events, serum creatinine, total GFR, and blood pressure (BP). After baseline evaluation, 26 of 66 patients underwent renal stenting because of injuries to the heart, brain, or kidneys. After 21 months, 6 medical patients required renal stenting, and 5 patients experienced late clinical events (2 medical patients, 3 stent patients). There was no difference in final BP between groups. Whereas medical patients experienced 6% and 8% decreases in total and stenotic kidney GFR, stent patients experienced 7% and 11% increases in total kidney (p = 0.006) and stenotic kidney (p = 0.02) GFR. There was no difference in final serum creatinine. In conclusion, patients with atherosclerotic RAS and baseline creatinine < or =2.0 mg/dl can be safely managed with aggressive medical therapy, with a small decrease in GFR. For patients who develop injuries to the heart, brain, or kidneys, renal artery stenting may further reduce hypertension and improve renal function. PMID- 16253608 TI - Chronotropic incompetence as a predictor of death among patients with normal electrograms taking beta blockers (metoprolol or atenolol). AB - Chronotropic incompetence, or an inability to increase heart rate during exercise, independently predicts death in patients not taking beta blockers. Whether it predicts death in patients taking beta blockers is not known. Consecutive patients (n = 3,736; mean age 58 +/- 11 years; 68% men), who were taking either metoprolol tartrate or atenolol and were referred for symptom limited exercise testing from 1990 to 2002 at a major academic medical center, formed the prospective study cohort. None had heart failure, pacemakers, atrial fibrillation, or any electrocardiographic abnormalities. Patients were followed for a median of 4.5 years for all-cause mortality. Chronotropic response was defined as the percentage of heart rate reserve used. A value of < or =62%, which was noted in 813 patients (22%), was considered abnormal, meaning that chronotropic incompetence was present. There were 173 deaths. After adjusting for age, gender, heart rate at rest, standard risk factors, other medications, Duke treadmill score, and heart rate recovery, chronotropic incompetence predicted death (adjusted hazard ratio 1.94, 95% confidence interval 1.43 to 2.64, p <0.0001). The association of chronotropic incompetence with death was present, irrespective of which drug was taken or the number of half-lives that had elapsed since the last dose. In conclusion, in patients taking beta blockers, chronotropic incompetence is independently predictive of death. PMID- 16253609 TI - Efficacy and safety of sildenafil added to treprostinil in pulmonary hypertension. AB - Pulmonary arterial hypertension (PAH) is characterized by abnormalities in endothelial and smooth muscle cell function. Prostacyclin released by endothelial cells is a potent vasodilator by increasing cyclic adenosine monophosphate. Sildenafil, an inhibitor of phosphodiesterase-5, increases cyclic guanosine monophosphate in the lungs, producing vasodilation. To test for a therapeutic benefit of the combination of a prostacyclin analogue, subcutaneous treprostinil, and sildenafil, a proof-of-concept, open-label investigational trial was initiated. Subjects with PAH in World Health Organization (WHO) functional classes II to IV receiving subcutaneous treprostinil for > or =6 months were evaluated with an exercise treadmill test using the Naughton-Balke protocol at baseline and after 12 weeks. Sildenafil 50 mg 3 times daily was added to the treprostinil. Mean treadmill times in seconds were compared before and after 12 weeks of therapy. Nine subjects enrolled in the trial; 7 were women (mean age 35 years). At baseline, 3 subjects were in WHO functional class II and 6 subjects were in WHO functional class III. The mean treadmill time at baseline was 465 +/- 167 seconds and at 12 weeks was 656 +/- 205 seconds (42% improvement, p = 0.049). All patients had symptomatic improvement. In conclusion, this pilot study of subcutaneous treprostinil with sildenafil for PAH suggests additive beneficial effects. PMID- 16253610 TI - Relation of brachial artery flow-mediated vasodilation to significant coronary artery disease in patients with peripheral arterial disease. AB - In patients at risk for coronary atherosclerosis, brachial artery flow-mediated dilation (FMD) rules out significant coronary artery disease (CAD). However, the value of this approach is unknown in patients with peripheral arterial disease who are at increased risk for CAD. This study assessed whether the noninvasive evaluation of endothelial function by brachial artery FMD rules out significant CAD by dipyridamole myocardial perfusion imaging (MPI) in patients with peripheral arterial disease who are asymptomatic for CAD. Forty-four patients with peripheral arterial disease who were asymptomatic for CAD underwent, in the same day, FMD evaluation and dipyridamole MPI using technetium-99m sestamibi single photon-emission computed tomography. MPI results were abnormal in 17 of 44 patients (39%). FMD was significantly less (6.0 +/- 2.3%) in patients with abnormal MPI results compared with those with normal MPI results (7.3 +/- 1.8%, p = 0.04). By multivariate analysis, FMD was the only significant predictor of abnormal MPI results (odds ratio 0.63, p = 0.02). Receiver-operating characteristic curve analysis assessing the ability of FMD to identify patients with summed stress scores > or =3 yielded an area under the curve of 0.74 (p = 0.009). A FMD value >6% provided 92% negative predictive power to rule out abnormal MPI results, with sensitivity of 79% and specificity of 73%. In conclusion, the noninvasive evaluation of endothelial function by FMD has high negative predictive accuracy and good sensitivity and specificity to detect abnormal MPI results in patients with peripheral arterial disease. Thus, it may represent a valuable screening test to rule out significant CAD in these patients. PMID- 16253611 TI - Predictors of left atrial spontaneous echocardiographic contrast or thrombus formation in stroke patients with sinus rhythm and reduced left ventricular function. AB - The objective of the present study was to identify predictors of left atrial spontaneous echocardiographic contrast (SEC) or thrombus in patients with stroke with sinus rhythm and left ventricular dysfunction. Of 500 consecutive patients with stroke, 48 with sinus rhythm and reduced left ventricular ejection fractions (EFs) < or =45% were examined. Ten patients presented with SEC or thrombus. The patients with SEC or thrombus had larger left atrial diameters (47 +/- 4 vs 42 +/ 6 mm, p <0.05), smaller EFs (30 +/- 9% vs 38 +/- 8%, p <0.01), and slower left atrial appendage (LAA) flow velocities (42 +/- 13 vs 61 +/- 17 cm/s, p <0.01). Multivariate analysis identified EF < or =35% and LAA flow velocity < or =55 cm/s as predictors of SEC or thrombus (p <0.05). Patients with stroke with sinus rhythm and moderate- to high-grade reduction of the left ventricular EF represent a risk group for a left atrial source of embolism and should undergo transesophageal echocardiography. PMID- 16253612 TI - Intrasubject variability of radial artery flow-mediated dilatation in healthy subjects and implications for use in prospective clinical trials. AB - Flow-mediated dilation (FMD) in the brachial artery is a widely used end point in clinical trials despite large within-subject variability and a small dynamic range. Recent studies suggest that the radial artery may be more advantageous for investigating FMD because of an enhanced vasodilator response. This study therefore assessed the validity and repeatability of radial artery FMD (FMD-R) to evaluate its suitability for the noninvasive evaluation of endothelial function. Thirty-three healthy subjects were recruited over a period of 11 months. Intra- and inter-reader reproducibilities were measured with high-resolution ultrasound at 4 time points: twice 1 morning (1 to 3 hours apart) and twice again within 7 days (range 4 to 9 days between visits). Conduit endothelial-dependent and independent vasomotion were assessed by responses to reactive hyperemia and nitroglycerin, respectively. FMD-R measurements demonstrated significant intra- and interday variabilities (intraclass correlation coefficients [ICCs] 0.38 and 0.23, p = 0.04 and 0.12, respectively). Bland-Altman plots confirmed the test retest variation in FMD-R. In contrast, radial artery diameter measurements (intra- and inter-reader) demonstrated a high degree of repeatability (interstudy ICC >0.8, p <0.0001). The number of subjects needed to detect a treatment difference of 2% in FMD-R with a p value of 0.05 and a power of 0.80 would be 118 in a crossover design and 234 in a parallel design for assessing group changes. In conclusion, these findings show that FMD-R is highly variable within subjects, even in a healthy population, after adjusting for multiple technical factors and implies biologic variation in radial artery tone. PMID- 16253613 TI - Studies on the intracellular localization of hHR23B. AB - Yeast Rad23, originally identified as a DNA repair protein, has been proposed to participate in other cellular functions, i.e., the proteasome-degradation pathway, the process of spindle pole body duplication and as a component of the anaphase checkpoint. Two human homologs of yeast Rad23, hHR23A and hHR23B, exhibit high sequence homology with yRad23 and also have been shown to be involved in DNA repair and proteasome-dependent degradation. Previous studies on the intracellular localization of hHR23A and hHR23B revealed their predominant localization in the nucleus during interphase and in the cytoplasm during mitosis. We have analyzed the localization of hHR23B during all the phases of the cell cycle using immunofluorescence. Unlike previous studies, our results suggest localization of hHR23B in the nucleus as well as in the cytoplasm during G1 phase. The nuclear levels of hHR23B decrease during S-phase of the cell cycle. When the cell enters mitosis, hHR23B relocalizes in the cytoplasm without association with chromatin. These results indicate that the intracellular distribution hHR23B is cell cycle dependent. PMID- 16253614 TI - Nobiletin and its related flavonoids with CRE-dependent transcription-stimulating and neuritegenic activities. AB - cAMP response element (CRE) transcription is dysregulated in neurodegenerative disorders in the central nervous system (CNS), including polyglutamine diseases. As the first step to find natural compounds with protective action against neurodegeneration in the CNS, we here examined whether six citrus flavonoids, namely nobiletin, 5-demethylnobiletin, tangeretin, sinensetin, 6 demethoxytangeretin, and 6-demethoxynobiletin, stimulated CRE-dependent transcription and induced neurite outgrowth in PC12D cells. Among the compounds, nobiletin most potently enhanced CRE-dependent transcription and neurite outgrowth by activating ERK/MAP kinase-dependent signalling to increase CREB phosphorylation. The transcription and neurite outgrowth were stimulated by nobiletin in a concentration-dependent manner, with a strong correlation between them. Furthermore, a 11-day oral administration of nobiletin rescued impaired memory in olfactory-bulbectomized mice documented to be accompanied by a cholinergic neurodegeneration. These results suggest that nobiletin with the activity to improve impaired memory may become a potential leading compound for drug development for neurodegenerative disorders exhibiting the dysregulated CRE dependent transcription. PMID- 16253615 TI - Thyroid autoimmunity, depression and anxiety; are there any connections? An epidemiological study of a large population. AB - OBJECTIVE: The aim of the study was to examine the relationship between antithyroid antibodies, depression, and anxiety in a large population. METHODS: In a population of 30,175 individuals aged 40-84 years, thyroid-stimulating hormone (TSH) was assessed in all women and in 50% of the men. Thyroid peroxidase autoantibodies (TPOAb) were measured in almost all samples with TSH > or = 4.0 mU/L (n = 1700) and in randomly selected samples without thyroid disease or biochemical dysfunction (n = 745). The levels of anxiety and depression were screened using the Hospital Anxiety and Depression Scale (HADS). Relations were investigated with multiple logistic regression analyses. RESULTS: In individuals with normal TSH and without known thyroid disorder, the prevalence of TPOAb was 14.2% in women and 4.3% in men. The prevalence of TPOAb in participants with biochemical thyroid dysfunction was 59.0% in women and 38.9% in men. No associations were found between thyroid autoimmunity and depression or anxiety, neither crude nor adjusted for age, gender, TSH, and thyroxine (T4). CONCLUSION: Thyroid autoimmunity is a common disorder in the population, mainly affecting females. In a population-based study, no associations were found between antithyroid antibodies and depression or anxiety. PMID- 16253616 TI - Do depression symptoms predict seizure frequency--or vice versa? AB - OBJECTIVE: The aim of this study was to test a theoretical explanatory model of the relationship between depression symptom scores and seizure frequency in people with epilepsy. METHODS: A community-based sample of adults with active epilepsy provided information on depression symptom scores and seizure frequency at two time points, 1 year apart. RESULTS: One thousand two hundred ten patients completed the initial questionnaire, and 976 of these individuals (80.7%) completed the final questionnaire. Depression scores and seizure frequency were significant predictors of each other, both within (beta = .07, P < .05 and beta = .09, P < .05) and across time (beta = .03, P < .01 and beta = .07, P < .05). CONCLUSION: The relationship between depression symptom scores and seizure frequency in those with epilepsy is bidirectional. PMID- 16253617 TI - A pilot study of the effects of behavioral weight loss treatment on fibromyalgia symptoms. AB - OBJECTIVE: Previous studies have found a relation between weight loss and pain severity in various chronic pain populations. However, there has been little research examining the relation between body mass index (BMI) and fibromyalgia syndrome (FMS). The purpose of this pilot study was to investigate the relationship between BMI and FMS symptoms and to determine if FMS symptoms would decrease following weight loss. METHODS: Overweight and obese women participated in a 20-week behavioral weight loss treatment. RESULTS: Participants, on average, lost 9.2 lbs (4.4% of their initial weight), and there were significant pre postimprovements on several outcome measures. Although weight was not significantly related to pain at baseline, weight loss significantly predicted a reduction in FMS, pain interference, body satisfaction, and quality of life (QOL). CONCLUSION: Findings suggest that behavioral weight loss treatment could be included in the treatment for overweight/obese women with FMS. PMID- 16253619 TI - Fatigue and psychiatric illness in a large community sample of persons with multiple sclerosis. AB - BACKGROUND: Fatigue and depression are among the most common and disabling symptoms in multiple sclerosis (MS), but the nature and extent of the relationships between fatigue and psychiatric disorders in MS patients remain poorly understood. METHODS: A mail survey was completed by 739 members of the King County (WA) MS Association. Fatigue was evaluated by questions from the Modified Fatigue Impact Scale (MFIS), depressive symptoms by the Center for Epidemiologic Studies Depression Scale (CES-D), substance-use disorders from the PRIME-MD. Information on demographics, employment and characteristics of MS was also collected. Logistic regression was used to identify covariates significantly associated with disabling fatigue. Receiver operating characteristic (ROC) curve analysis evaluated the sensitivity and specificity of fatigue for depression. RESULTS: Twenty-five percent of this community-dwelling sample reported that their activities were often or almost always limited by fatigue. Seventy-six percent of subjects with disabling fatigue had significant depressive symptoms (CES-D >16), compared with 31% of those without disabling fatigue. Depression was strongly associated with fatigue, after controlling for age, gender, marital status, and severity, course and duration of illness. In logistic regression analysis, subjects with clinically significant depressive symptoms (CES-D >16) were much more likely to report disabling fatigue: OR = 6.24 (4.16, 9.35). Anxiety and substance-use disorders did not have the same strong associations with fatigue. Fatigue was highly sensitive and specific for clinically significant depressive symptoms. CONCLUSIONS: Disabling fatigue is strongly associated with clinically significant depressive symptoms. Patients who report disabling fatigue should be screened for depression. PMID- 16253618 TI - The prevalence of self-reported chronic fatigue in a U.S. twin registry. AB - OBJECTIVE: To investigate the prevalence and correlates of various definitions of self-reported lifetime fatiguing illness in a U.S. twin registry. METHODS: Data from 4591 female and male twins from the population-based Mid-Atlantic Twin Registry were available for this study. Variables representing different definitions of lifetime fatiguing illness and personal characteristics were obtained through questionnaires. Odds ratios and 95% confidence intervals were calculated as measures of association between fatigue and gender. Kaplan-Meier curves were produced to examine the age at onset for lifetime fatiguing illnesses. RESULTS: Prevalences for different definitions of self-reported lifetime fatigue ranged from 36.7% for any fatigue to 2.7% for chronic fatigue syndrome-like illness. Females were two to three times more likely to report fatigue than males. Gender differences increased as fatigue definitions grew more restrictive. Ages at onset of chronic fatiguing illness were significantly earlier and the number of ancillary symptoms was greater for females than males. People with lifetime fatigue had significantly more compromised functional status than people without lifetime fatigue. CONCLUSION: The prevalence of self-reported lifetime fatiguing illness varied widely depending upon how it was defined. Given the debilitating consequences of fatiguing illnesses, the reasons for the female predominance and the earlier onset in women should receive increased research priority. PMID- 16253620 TI - Dependable social relationships predict overall survival in Stages II and III breast carcinoma patients. AB - OBJECTIVE: The effect of support, from dependable non-household relationships, on breast cancer progression was studied using a prospective, longitudinal design. METHODS: Dependable social support was assessed in women with invasive breast carcinoma Stages II and III within 18 months after diagnosis. Disease outcome was monitored for 8 to 9 years. Cox regression analyses, including the Nottingham Prognostic Index (NPI) score of disease severity, tested the association between the number of dependable support persons and time to death. RESULTS: Ninety participants listed between 1 and 16 dependable nonhousehold support persons (mean = 6, S.D. = 4). There were 21 recurrences and 16 deaths from breast cancer at the end of the study. The number of dependable supports predicted decreased mortality [RR = 0.41 (0.21-0.80), P = .01] after controlling for NPI. NPI predicted increased mortality [RR = 1.6 (1.0, 2.4), P = .05]. CONCLUSIONS: For patients with Stages II and III breast cancer, the number of dependable, nonhousehold relationships predicts decreased mortality, after accounting for disease severity. PMID- 16253621 TI - Support providers of sexual minority women with breast cancer: who they are and how they impact the breast cancer experience. AB - OBJECTIVE: The aim of this study was to identify the factors associated with adjustment to breast cancer among sexual minority women with breast cancer and their support person. METHODS: In a cross-sectional study, sexual minority women with breast cancer and their support provider were asked to self-report social support, distress, and coping, using standardized measures. RESULTS: Twenty-three (77%) women had a support provider participating in the study. Disclosure of sexual orientation, less helpless-hopeless coping, and support provider perception of high fighting spirit were related to lower patient distress. Lower support provider distress was related to more patient disclosure of sexual orientation, a larger social network, and an underestimation of fatalistic patient coping. An overestimation of patients' anxious preoccupation coping was linked to higher support provider distress. CONCLUSIONS: Providing opportunities to sexual minority patients and their support providers to focus on issues such as disclosure of sexual orientation and coping may lower patient and support provider distress. PMID- 16253622 TI - Do illness perceptions predict attendance at cardiac rehabilitation and quality of life following myocardial infarction? AB - OBJECTIVE: The aim of this study was to examine the extent to which illness perceptions predict attendance at cardiac rehabilitation and quality of life following myocardial infarction (MI). METHODS: The illness perceptions of 194 MI patients were assessed whilst the patients were still in hospital following an MI. The mean age was 63.3 years (S.D. = 10.6), and 142 of the patients were men. Cardiac rehabilitation attendance and quality of life were assessed via a postal questionnaire 6 months later. RESULTS: In contrast to previous work reported in this area, illness perceptions were not significantly associated with attendance at cardiac rehabilitation. Illness perceptions measured within 24 h of an acute MI were predictive of quality of life 6 months later. CONCLUSION: Previous reports may have overestimated the extent to which illness perceptions predict attendance at cardiac rehabilitation. The relationship between illness perceptions and quality of life at 6 months suggests that interventions to alter illness perceptions, especially perceptions of consequences, may be useful in improving health-related quality of life (HRQoL) following an MI. PMID- 16253623 TI - Does it matter what patients think? The relationship between changes in patients' beliefs about angina and their psychological and functional status. AB - OBJECTIVE: The aim of this study is to examine the association between changes in misconceived or maladaptive beliefs about angina and patients' functional and psychological status. METHOD: The method used was a prospective follow-up study over 1 year of 133 people with angina. RESULTS: Beliefs about angina were significantly associated with functional and psychological status. People with more misconceived or maladaptive beliefs were more anxious and physically limited than were people with fewer such beliefs, with differences in physical functioning that were clinically significant. Change in angina beliefs over 1 year was the most significant predictor for physical functioning at follow-up, after controlling for the effects of demographic variables and the outcome variable at baseline, whereas change in the frequency of angina did not contribute significantly to this model. CONCLUSION: Misconceived and maladaptive beliefs about angina are associated with reductions in both functional and psychological status. These beliefs are easily and quickly identified using a simple questionnaire and should be corrected. PMID- 16253624 TI - Childhood adversities experienced by working-aged coronary heart disease patients. AB - OBJECTIVE: The aim of this study is to investigate associations between childhood adversities and coronary heart disease (CHD). METHODS: This was a case-control study based on a postal questionnaire addressed to randomly selected working-aged Finns, and response rate was 39% (N = 15,477). The sample comprised 319 CHD patients. Four age- and gender-matched controls were selected for every patient. The participants were asked in six questions to think about their childhood adversities. RESULTS: Fear of some family member and someone in the family being seriously or chronically ill were more common during childhood among working-aged CHD patients than among controls. Likewise, among female CHD patients, serious conflicts in the family and someone in the family having had alcohol problems and, among male CHD patients, long-lasting financial problems were more common than among controls. Odds ratios (OR) varied between 1.27 and 2.66. Adjustment for education had no influence among women, but it had an influence among men. Upon adjustment for conventional risk factors (smoking, obesity, and hypertension), the association mostly disappeared. A family member having been seriously or chronically ill was statistically significant after full adjustment among both genders. CONCLUSION: Working-aged CHD patients have experienced more dramatic events during their childhood than did the control population. This issue cannot be solved in doctors' offices. Health-promoting social policies are of vital importance. PMID- 16253625 TI - Coping and survival in patients with leukemia undergoing allogeneic bone marrow transplantation--long-term follow-up of a prospective study. AB - OBJECTIVE: The exploratory study examined the relationship between coping and survival in patients undergoing bone marrow transplantation (BMT). METHODS: Patients scheduled for BMT were recruited from 1990 until 1995 at the University Hospital of Ulm, Germany. They were interviewed before transplantation, and the corresponding records were checked in December 2002. Seventy-two audiotaped interviews could be analyzed for 34 coping strategies as defined in the Ulm Coping Manual (UCM). Main outcome measure was survival time post-BMT. RESULTS: On average, the patients were 35 years old, 65% were male, and 56% diagnosed acute leukemia (AL). Four coping strategies were found to show a clear trend towards an association with survival time: emotional support, acceptance, taking control, and compensation. The last strategy was associated with shorter, the others with longer survival. CONCLUSION: We found further evidence for an association between coping and survival. Because of the possible wide-reaching consequences for clinical management, replication of the data is essential. PMID- 16253626 TI - Invited commentary to the paper "Coping and survival in patients with leukemia undergoing allogeneic bone marrow transplantation--long term follow-up of a prospective study" by Norbert Grulke et al. (OJPR/2004/3520). PMID- 16253627 TI - Substrate oxidation during exercise at moderate and hard intensity in middle-aged and young athletes vs sedentary men. AB - This study investigated the combined effects of endurance training and aging on substrate oxidation during exercise. Thirty-one healthy male subjects in 4 groups (8 middle-aged trained cyclists, 8 young trained cyclists, 7 middle-aged sedentary men, and 8 young sedentary men) performed 2 50-minute cycle ergometer exercise tests, below and above (+/-15%) their individual ventilatory threshold (VT). Substrate oxidation was evaluated by indirect calorimetry during the steady state tests. Aging decreased carbohydrate (CHO) use (P<.05) in all subjects regardless of fitness status or exercise intensity. However, it declined 2-fold less in the trained men (P<.05) and was associated with a stronger epinephrine response (P<.05). During hard-intensity exercise, endurance training increased by 100% CHO use in the older men (P<.05). In the younger men, training increased fat oxidation but did not change CHO oxidation, resulting in a marked decrease in the ratio between CHO and fat used at high-intensity exercise (-93%; P<.05). These data suggest an age-related decline in the use of CHO as an energy source in exercising men, independent of intensity level. This decline, however, is attenuated in well-trained men for exercise intensities above the VT. In view of these findings, we hypothesize that cycling training performed at a specific exercise intensity (ie, 15% above VT) may improve CHO mobilization and use in middle-aged men. PMID- 16253628 TI - Effect of impaired glucose uptake on postexercise glycogen repletion in skeletal muscles of insulin-treated streptozotocin-diabetic fasted rats. AB - During recovery from intense exercise performed while fasting, the replenishment of muscle glycogen stores from glucose requires the activation of glucose transport. This study examines if insulin-treated streptozotocin (STZ) diabetes in rats impairs the rate of muscle glucose utilization and glycogen repletion when no food is ingested during recovery from high-intensity exercise. Rats fasted for 24 hours were injected with high doses of STZ (150 mg/kg) to cause severe diabetes, and their glycemia was normalized for 10 days with twice-daily insulin injections. High-intensity exercise in these rats resulted in a marked increase in plasma glucose, which remained higher than preexercise levels thereafter, whereas in control animals, the rise in glycemia was only of a short duration. During recovery, the rates of 2-deoxy-[(3)H]glucose utilization in muscles rich in fast twitch red fibers (red and mixed gastrocnemius muscles) were much lower in STZ-diabetic than in control rats, but were not affected by diabetes in muscles comprised mainly of fast twitch white fibers (white gastrocnemius muscle). Despite these effects on glucose utilization, STZ diabetes had no inhibitory effect on the rate and extent of glycogen deposition and fractional velocities of glycogen synthase across all muscles. In conclusion, although insulin-treated STZ diabetes in fasted rats inhibits glucose transport rates in fast twitch red muscle fibers post-intense exercise, this has no effect on muscle glycogen repletion either because glucose transport does not control the rate of glycogen synthesis or because of a compensatory increase in the activity of lactate glyconeogenesis in these muscles. PMID- 16253630 TI - Genome-wide search for genes affecting serum uric acid levels: the Framingham Heart Study. AB - Serum uric acid levels are associated with hypertension, cardiovascular disease, and renal disease. Uric acid has been shown to be heritable; however, genome-wide linkage analyses have not been reported. Genome-wide multipoint variance components linkage analyses with 401 markers spaced at approximately 10 centimorgan (cM) were conducted on 1258 subjects of the Framingham Heart Study, using the average of two serum uric acid measurements obtained in examinations 1 and 2 around 1971 and 1979. Covariates in fully adjusted model included sex, age, body mass index (BMI), serum creatinine, alcohol consumption, diabetes, diuretic treatment, and triglycerides. To investigate possible pleiotropic effects between uric acid and covariates that may have a genetic component, bivariate linkage analyses of uric acid with BMI, triglycerides, and glucose were conducted at the uric acid linkage regions. The heritability of uric acid was 0.63. The highest multipoint log-of-the-odds (LOD) score was 3.3 at 50 cM on chromosome 15 for age sex-adjusted uric acid, but decreased to 1.5 after multivariable adjustment. Additional evidence of linkage was seen on chromosomes 2 (LOD score 1.1 at 4 cM) and 8 (LOD score 1.7 at 6 cM) for multivariable-adjusted uric acid. Pleiotropic effects were only found between uric acid and glucose and BMI at chromosomes 8 and 15 linkage locations, respectively. We have identified several novel loci linked to uric acid. We found possible pleiotropic effects between uric acid and BMI and glucose. Further research is necessary to identify the genes involved in uric acid metabolism and their roles in hypertension, cardiovascular disease, and renal disease. PMID- 16253629 TI - Labeled CO(2) production and oxidative vs nonoxidative disposal of labeled carbohydrate administered at rest. AB - Carbon isotopes (*C) have been extensively used in man to describe oxidative vs nonoxidative disposal of an exogenous load of labeled carbohydrate (*C-CHO) at rest in various experimental situations. It is hypothesized that V*CO(2) reflects *C-CHO oxidation. However, when glycogen is synthesized through the indirect pathway (which is responsible for approximately 50% of glycogen storage), *C could be lost, diluted, and exchanged in the pyruvate-lactate pool, in the pool of tricarboxylic acid cycle intermediates, as well as at the entrance of the tricarboxylic acid cycle, and along the pathway of gluconeogenesis. This could result in a lower *C/C in the glycogen stored than in the CHO administered, in an increased production of *CO(2), and, respectively, in an overestimation and an underestimation of the oxidative and nonoxidative disposal of the CHO load. Results from the present experiment offer a support to this hypothesis. Over a 10 hour period after ingestion of a (13)C-pasta meal (313+/-10 g dry mass or 258+/-8 g of glucose) in 12 healthy subjects (6 men and 6 women), exogenous CHO oxidation computed from V(13)CO(2) (recovery factor, 0.54) significantly exceeded total CHO oxidation computed by indirect respiratory calorimetry corrected for urea excretion: 154.2+/-2.6 vs 133.5+/-3.2 g. In an additional study conducted in rats, (13)C/(12)C in glycogen stores was significantly approximately 50% lower than in the (13)C-CHO ingested, over a wide range of enrichment. These results suggest that because of dilution, loss, and exchange of *C in the indirect pathway of glycogen synthesis, the oxidative vs nonoxidative disposal of exogenous *C-CHO cannot be accurately tracked from V*CO(2). PMID- 16253631 TI - Fasting apoprotein B-48 level and coronary artery disease in a population without frank fasting hypertriglyceridemia. AB - The aim of this study was to test the hypothesis that fasting apoprotein B-48 level might be a surrogate marker of postprandial lipemia in evaluating the risk of coronary artery disease (CAD) in a population without frank abnormality in fasting lipid profile. One hundred twenty-three patients tested by coronary angiography were selected on the criteria of absence of treatment with hypolipidemic drugs, obvious hypertriglyceridemia (>2.85 mmol/L), or other conditions that may interfere with lipoprotein metabolism except diabetes. CAD was defined by more than 50% narrowing of vessel lumen, and its severity is determined by the number of arteries involved. Fasting apoprotein B-48 was measured by a competitive enzyme-linked immunosorbent assay method. There was no difference in fasting apoprotein B-48 levels between the groups with and without CAD (0.123+/-0.096 vs 0.136+/-0.125 microg/mL, respectively), whatever the sex or whether with or without diabetes. The apoprotein B-48 level was not related to the presence or the severity of CAD. There was also no correlation between fasting apoprotein B-48 levels and age, sex, body mass index, and usual fasting lipid parameters in both patients with and without angiographically proven CAD. Finally, among the features of metabolic syndrome, apoprotein B-48 was correlated with fasting triglyceride levels (r=0.357, P<.01) only. In conclusion, the present study shows that in the absence of any major fasting abnormality in plasma lipid parameters, fasting apoprotein B-48 level, which has been associated with postprandial hyperlipidemia, does not predict the risk of CAD. PMID- 16253632 TI - A white blood cell count in the normal concentration range is independently related to cardiorespiratory fitness in apparently healthy Korean men. AB - Despite the documented health benefits of physical activity, the mechanism whereby physical activity prevents cardiovascular disease is incompletely understood. In the present study, we investigated the relationship between white blood cell (WBC) count and cardiorespiratory fitness (V o(2)max) after adjusting for several well-known cardiovascular risk factors. Subjects who visited our health promotion center for a medical checkup and treadmill test (n=8241; age: median, 48 years; range, 16-79 years) were classified into 3 groups based on their WBC counts (group 1, 2200-5300 microL, n=2823; group 2, 5301-6500 microL, n=2709; group 3, 6501-10000 microL, n=2709). After adjusting for age, body mass index, body fat percentage, smoking history, systolic blood pressure, diastolic blood pressure, serum lipid profile, and fasting plasma glucose, V o(2)max still showed a significant association with WBC count (partial r=-0.11, P<.001). In logistic regression analyses, subjects in the highest WBC tertile showed lower V o(2)max compared with those in the lowest WBC tertile after adjusting for age and cardiovascular risk factors (odds ratio, 0.42; 95% confidence interval, 0.36-0.49 for the highest V o(2)max tertile). These results suggest that a WBC count in the normal concentration range is independently related to cardiorespiratory fitness in Korean men. PMID- 16253633 TI - Elevated glucose activates protein synthesis in cultured cardiac myocytes. AB - Diabetes mellitus results in chronic hyperglycemia, a serious metabolic disorder associated with a markedly increased risk of cardiovascular disease. However, the effects of high glucose (HG) on cardiac myocyte growth have not been fully clarified. In this study, the effect of glucose on cardiac myocyte growth was examined using leucine incorporation as an index of protein synthesis. High glucose (HG, 25 mmol/L) increased leucine incorporation (167% +/- 0.2% over normal glucose, n=4, P<.01) compared with a physiological glucose concentration (5.5 mmol/L, normal glucose). The HG-induced increase in leucine incorporation was time- and dose-dependent and was not due to osmotic changes because 25 mmol/L mannitol did not change leucine incorporation. High glucose also significantly reduced elongation factor 2 phosphorylation, an effect known to result in increased protein synthesis at the elongation step. Western blot analysis showed that HG-activated protein kinase B (PKB), also called Akt (PKB/Akt), at 18 hours. High glucose-induced leucine incorporation was attenuated with phosphatidylinositol 3-kinase (PI3K) inhibition using wortmannin and LY294002 and by rapamycin, a mammalian target of rapamycin (mTOR) inhibitor, 72%, 64%, and 65% (P<.05), respectively. High glucose also activated extracellular signal-regulated kinase 1/2 activity with peak stimulation at 5 minutes. In addition, PD98059, an inhibitor of mitogen-activated protein kinase kinase, attenuated HG-induced leucine incorporation. These data show for the first time that elevated glucose increases protein synthesis in cardiac myocytes. The increase appears to be mediated by activation of PI3K-PKB/Akt and/or PI3K-mTOR as well as extracellular signal-regulated kinase 1/2. These results provide new evidence for a direct effect of glucose independent of insulin on cardiac myocyte growth. PMID- 16253634 TI - Elevation of serum adiponectin levels in Basedow disease. AB - The present study was undertaken to determine whether thyroid hormone affects serum adiponectin levels in the patients with Basedow disease. Sixty-four patients with Basedow disease were examined; 32 patients had hyperthyroid state and 32 patients had euthyroid state who had been treated with antithyroid drugs. In addition, 30 age- and sex-matched subjects served as a control. Serum adiponectin, free T4, free T3, thyroid-stimulating hormone, and thyroid stimulating hormone receptor antibody (TRAb) were measured. Serum adiponectin levels were 12.9+/-1.6 microg/mL in the hyperthyroid state, a value significantly greater than that of 8.2 +/- 0.5 microg/mL in the euthyroid state (P<.05) and that of 8.6+/-0.7 microg/mL in the control subjects (P<.05). Serum adiponectin levels had positive correlations with either of serum free T4 (r=0.453, P<.001), free T3 (r=0.47, P< .001), or TRAb (r= 0.491, P<.001), but not with body mass index. Multiple regression analysis showed TRAb had the strongest contribution to serum adiponectin concentration in the patients with Basedow disease. The present findings indicate that hyper-adiponectinemia is closely associated with increases in serum thyroid hormone levels and TRAb in Basedow disease. PMID- 16253635 TI - Hypercholesterolemia, not metabolic syndrome, related to adhesion of monocytes to cultured endothelium in nondiabetic subjects. AB - The interaction of leukocytes and endothelium plays an important role in the development of atherosclerosis. Previous studies found that adhesion of leukocytes to endothelium is greater in subjects with hypercholesterolemia. It is not clear if metabolic syndrome, a contributing risk factor of cardiovascular disease, is related to this adhesion. Therefore, we conducted a study, in which 48 nondiabetic subjects were enrolled, to determine the relationship between leukocyte adhesion and the components of metabolic syndrome. After a 12-hour overnight fast, subjects' fasting blood was obtained for measurement of lipoprotein concentrations and glucose and insulin levels. Results of the number of monocyte adhesion to human umbilical vein endothelial cells were divided into high monocyte adhesion group and low monocyte adhesion group (n=24 in each group). Plasma concentrations of total cholesterol (245 +/- 5 vs 229+/- 4 mg/dL, P=.021) and low-density lipoprotein cholesterol (LDL-C) (162+/- 4 vs 146+/- 3 mg/dL, P=.003) were both higher in the high monocyte adhesion group than in the low monocyte adhesion group. Monocyte adhesion was significantly correlated to plasma concentrations of LDL-C (r=0.407, P=.002) but not to the total cholesterol (r=0.202, P=.085). However, there was no difference in monocyte adhesion to endothelium between subjects with or without metabolic syndrome, based on the modified criteria from the Adult Treatment Panel III of the National Cholesterol Education Program. Insulin resistance index, presented as homeostasis model assessment insulin resistance, and glucose or insulin responses to oral glucose tolerance test were similar between groups. Our study demonstrated that monocyte adhesion to endothelium has a stronger relationship with the plasma concentration LDL-C than with characteristics of metabolic syndrome. PMID- 16253636 TI - Aerobic exercise training improves insulin sensitivity without changes in body weight, body fat, adiponectin, and inflammatory markers in overweight and obese girls. AB - The aim of this study was to examine the effect of aerobic exercise training on insulin sensitivity in overweight and obese girls. Nineteen overweight and obese girls (mean +/- SD: age, 13.1+/-1.8 years; body mass index, 26.8+/-3.9 kg/m(2)) volunteered for this study. Body composition (dual-energy x-ray absorptiometry), insulin sensitivity (oral glucose tolerance test and homeostasis model assessment estimate of insulin resistance; n=15), adiponectin, C-reactive protein (CRP), interleukin (IL) 6, insulin-like growth factor-1, soluble intercellular adhesion molecule-1 and soluble vascular cell adhesion molecule-1 serum levels, and blood lipids and lipoproteins were assessed before and after 12 weeks of aerobic training. Cardiorespiratory fitness increased by 18.8% (P<.05) as a result of training. The area under the insulin concentration curve (insulin area under the curve) decreased by 23.3% (12781.7+/-7454.2 vs 9799.0+/-4918.6 microU.min/mL before and after intervention, respectively; P=.03). Insulin sensitivity was improved without changes in body weight (pre-intervention, 67.9+/-14.5 kg; post intervention, 68.3+/-14.0 kg) or percent body fat (pre-intervention, 41.4% +/- 4.8%; post-intervention, 40.7%+/-5.2%). The lower limb fat-free mass increased by 6.2% (P<.01) as a result of training, and changes in lower limb fat-free mass were correlated with changes in the insulin area under the curve (r= -.68; P< .01). Serum adiponectin, IL-6, and CRP concentrations did not change (pre intervention vs post-intervention: adiponectin, 9.57+/-3.01 vs 9.08+/-2.32 microg/mL; IL-6, 1.67+/-1.29 vs 1.65+/-1.25 pg/mL, CRP, 3.21+/-2.48 vs 2.73+/ 1.88 mg/L) whereas insulin-like growth factor-1 was lower after training (pre intervention, 453.8 +/- 159.3 ng/mL; post-intervention, 403.2+/- 155.1 ng/mL; P<.05). In conclusion, 12 weeks of aerobic training improved insulin sensitivity in overweight and obese girls without change in body weight, percent body fat, and circulating concentrations of adiponectin, IL-6, CRP, and other inflammatory markers. These findings suggest that increased physical activity may ameliorate the metabolic abnormalities associated with obesity in children with a mechanism other than the parameters cited earlier. PMID- 16253637 TI - Raw vegetable food containing high cyclo (his-pro) improved insulin sensitivity and body weight control. AB - Cyclo (his-pro), controlled-energy diet, soy protein hydrolysate (SPH), and raw vegetable food (RVF) are known to improve insulin sensitivity and body weight (BW) control. Enhancement of high cyclo (his-pro) content in SPH (HCS) was performed by refluxing SPH with 1 N KH(2)CO(3) dissolved in 70% ethanol for 2 weeks at room temperature. Using this material, we examined the effects of HCS plus RVF on glucose metabolism and BW control in genetically diabetic Goto Kakizaki (G-K) and insulin-resistant aged overweight Sprague-Dawley (S-D) rats. Thirty 7-week-old G-K rats and 18 16- to 18-month-old S-D rats were divided into 3 groups and treated with normal chow (NC), RVF diet, or HCS diet for 8 weeks. Raw vegetable food diet was made of 1:3 RVF and 2:3 NC; HCS diet was made of 1:27 portion HCS, 8:27 RVF, and 2:3 NC. Oral glucose tolerance significantly improved in both RVF- (P<.01) and HCS-treated (P<.001) G-K rats and worsened in NC-fed rats compared with the baseline values. Similarly, oral glucose tolerance also improved in aged overweight S-D rats when treated with RVF (P<.05) and with HCS (P<.01), compared with the baseline values. Although HCS diet treatment very significantly lowered fed plasma insulin levels compared with NC diet treatment in G-K rats (P<.01), RVF diet treatment alone did not decrease plasma insulin levels. In contrast, there was no change of insulin levels in overweight aged S-D rats after either RVF or HCS diet treatment. Postfeeding glucose levels in G-K rats fed RVF or HCS significantly fell, compared with the rats fed NC (P<.05). Interestingly, fasting blood glucose levels in RVF- or HCS-fed rats were very significantly lower than in NC-fed rats (P<.001). There was no change of blood glucose levels in S-D rats due to treatments with different diet. In G-K rats, food intake did not decrease during the first 3 weeks but fell very significantly from the fifth to eighth weeks with RVF (P<.01) and HCS (P<.001) treatments in G K rats. However, food intake reduction in aged S-D rats was shown only for the HCS-treated rat group (P<.05). Water intake slightly decreased in G-K rats with either RVF or HCS treatment (P<.05) but very significantly decreased in S-D rats with HCS treatment (P<.01). Body weight gain in young G-K rats and BW in aged S-D rats significantly decreased only when rats were treated with HCS diet (P<.05). These data suggest that regular consumption of HCS diet helps to control blood glucose metabolism in diabetic G-K rats and BW control in aged obese S-D rats. PMID- 16253638 TI - Transgenic expression of mutant peroxisome proliferator-activated receptor gamma in liver precipitates fasting-induced steatosis but protects against high-fat diet-induced steatosis in mice. AB - Steatosis is one of the most common liver diseases and is associated with the metabolic syndrome. A line of evidence suggests that peroxisome proliferator activated receptor (PPAR) alpha and PPARgamma are involved in its pathogenesis. Hepatic overexpression of PPARgamma1 in mice provokes steatosis, whereas liver specific PPARgamma disruption ameliorates steatosis in ob/ob mice, suggesting that hepatic PPARgamma functions as an aggravator of steatosis. In contrast, PPARalpha-null mice are susceptible to steatosis because of reduced hepatic fatty acid oxidation. PPARgamma with mutations in its C-terminal ligand-binding domain (L468A/E471A mutant PPARgamma1) have been reported as a constitutive repressor of both PPARalpha and PPARgamma activities in vitro. To elucidate the effect of co suppression of PPARalpha and PPARgamma on steatosis, we generated mutant PPARgamma transgenic mice (Liver mt PPARgamma Tg) under the control of liver specific human serum amyloid P component promoter. In the liver of transgenic mice, PPARalpha and PPARgamma agonist-induced augmentation of the expression of downstream target genes of PPARalpha and PPARgamma, respectively, was significantly attenuated, suggesting PPARalpha and PPARgamma co-suppression in vivo. Suppression of PPARalpha and PPARgamma target genes was also observed in the fasted and high-fat-fed conditions. Liver mt PPARgamma Tg were susceptible to fasting-induced steatosis while being protected against high-fat diet-induced steatosis. The opposite hepatic outcomes in Liver mt PPARgamma Tg as a result of fasting and high-fat feeding may indicate distinct roles of PPARalpha and PPARgamma in 2 different types of nutritionally provoked steatosis. PMID- 16253639 TI - Carriers of the frequent lipoprotein lipase S447X variant exhibit enhanced postprandial apoprotein B-48 clearance. AB - The frequent lipoprotein lipase S447X variant (LPLS447X) is firmly associated with a lower incidence of cardiovascular disease, the mechanisms for which remain to be established. To further unravel these beneficial effects, we studied the consequences of LPLS447X heterozygosity on LPL mass and activity, as well as on the postprandial lipoprotein profile. Fifteen male heterozygous LPLS447X carriers and 15 matched control subjects received an oral fat load (30 g/m(2)). Lipid parameters were evaluated at baseline and 2, 3, 4, and 6 hours after fat loading. LPL concentration and activity were analyzed, and endothelial function was evaluated noninvasively as flow-mediated dilation of the brachial artery. Although baseline apoprotein B-48 (apoB48) levels were similar, the rise in apoB48 was attenuated in LPLS447X carriers with 25% lower peak values compared with controls (P=.04). In conjunction, LPLS447X carriers were characterized by a 2.4-fold increase in pre-heparin LPL mass (P<.0001). The decrease in postprandial flow-mediated dilation was comparable in both groups. LPLS447X carriers exhibit enhanced apoB48 clearance with concomitant increase in pre-heparin LPL mass, without changes in LPL activity. This combination might suggest a role for increased ligand action of LPL in LPLS447X carriers contributing to the cardiovascular protection in carriers of this mutation. PMID- 16253640 TI - Atherogenic lipoprotein subfraction profile in preeclamptic women with and without high triglycerides: different pathophysiologic subsets in preeclampsia. AB - Abnormal lipid metabolism has been proposed as a pathogenic factor of preeclampsia, although whether it is a constant feature in all preeclamptic patients is unclear. We assessed whether plasma triglyceride (TG) levels can distinguish a subgroup of preeclamptic women with alterations in lipoprotein profile from those with normal lipid metabolism and can be used to identify 2 distinct pathogenic groups in preeclampsia. This prospective study included 34 women with preeclampsia and 23 healthy pregnant women. Preeclamptic women were further subclassified into normal-TG (<250 mg/dL) and high-TG (>or=250 mg/dL) groups on the basis of the 90th percentile in our population. Low-density lipoproteins (LDLs) were ultracentrifuged and were separated into 4 subfractions, and lipid distribution in the subfractions was analyzed in all study groups. Vascular cell adhesion molecule-1 was also measured as a marker of endothelial dysfunction. Sixteen women with preeclampsia had high TGs (47% vs 13% in control subjects, P<.001). This subgroup showed a significant shift in lipid distribution, mainly, TGs, toward the small, dense LDL subfractions. However, preeclamptic patients in the normal-TG subgroup showed LDL subfraction lipid distribution similar to that of healthy pregnancies. Vascular cell adhesion molecule-1 levels were significantly elevated in preeclamptic patients in comparison with those in control subjects regardless of TG levels. The presence of a proatherogenic lipoprotein profile, previously described in preeclampsia, is characterized by increased small dense LDL and is exclusive to a subset of preeclamptic patients with high TG levels. These findings support the concept of heterogeneous pathogenic lines in preeclampsia and the use of subclassifications in pathophysiologic research on this condition. PMID- 16253641 TI - Association among serum ferritin, alanine aminotransferase levels, and metabolic syndrome in Korean postmenopausal women. AB - We examined the relationships among serum ferritin, alanine aminotransferase (ALT) levels, and cardiovascular risk factors of metabolic syndrome in Korean postmenopausal women. We conducted a cross-sectional study of 959 postmenopausal women without an apparent cause of liver disease. Metabolic syndrome was defined as the presence of at least 3 of the following: elevated blood pressure, low high density lipoprotein cholesterol, elevated serum triglycerides, elevated plasma glucose, and abdominal obesity. Serum ferritin and ALT levels were found to be correlated (r=0.374, P<.001) and to be associated with the components of metabolic syndrome. Subjects with metabolic syndrome showed significantly higher serum ferritin (74.7+/- 2.0 vs 59.6+/- 2.0 ng/mL, P<.001) and ALT levels (21.3+/ 1.6 vs 18.7+/-1.5 IU/L, P<.001). Moreover, the greater the number of metabolic syndrome components present, the higher were the serum ferritin and ALT levels (P<.001). Multiple regression analysis showed that serum ALT levels are significantly associated with serum ferritin levels, waist circumference, fasting blood glucose, age, and white blood cell count (adjusted R(2)=0.147). Elevated iron stores were positively associated with serum ALT levels and metabolic syndrome in Korean postmenopausal women. PMID- 16253642 TI - Endothelin antagonism improves hepatic insulin sensitivity associated with insulin signaling in Zucker fatty rats. AB - In the present study, we investigated the effects of long-term treatment with the endothelin (ET) antagonist atrasentan, an ET(A)-selective antagonist, on whole body glucose metabolism and insulin signaling in a commonly used model of insulin resistance, the Zucker fatty rat. Zucker lean and fatty rats were maintained for 6 weeks on either control or atrasentan-treated water. Euglycemic hyperinsulinemic clamps (4 mU/kg per minute) were performed at the end of the 6 week treatment on a subset of rats (n=10/treatment). In another subset (n=5/treatment), an insulin tolerance test was performed; liver and muscle tissues were harvested 10 minutes following the challenge for further analysis. Results of the clamps demonstrated that long-term atrasentan treatment significantly increased whole body glucose metabolism in fatty rats compared with vehicle control subjects. Insulin-induced insulin receptor substrate 1 tyrosine and protein kinase B serine phosphorylation were significantly reduced in the liver and muscle of fatty animals compared with their lean littermates. This reduction was overcome with atrasentan treatment in the liver but not in the muscle. There was no difference between lean and fatty animals, however, in insulin receptor substrate 1 and protein kinase B protein expression in the liver and muscle and no effect by atrasentan. In contrast, expression of the regulatory subunit of PI-3 kinase (p85alpha) was significantly increased in the liver but not in the muscle of fatty animals compared with their lean littermates and this was normalized to levels of lean animals with atrasentan treatment. These findings indicate that long-standing ET antagonism improves whole body glucose metabolism in Zucker fatty rats through improvements in insulin signaling in the liver. These results indicate that therapeutic ET antagonism may assist in correcting the insulin-resistant state. PMID- 16253643 TI - Lower serum free thyroxine levels are associated with metabolic syndrome in a Chinese population. AB - Thyroid hormones play an important role in regulating energy homeostasis and lipid and glucose metabolism. This study assessed the relationship between free thyroxine and clinical features of metabolic syndrome (MS). A total of 4,938 Taiwanese subjects (2,891 men and 2,047 women with a mean age of 50.1+/-12.6 years) with normal serum free thyroxine levels were enrolled. A modified National Cholesterol Education Program definition of MS was adopted substituting body mass index (BMI) for waist circumference. Serum free thyroxine concentrations were determined by immunoassay. Overall, 14% of subjects had a high fasting glucose, 27% had high blood pressure, 14% had high serum total triglyceride, 8% had low high-density lipoprotein cholesterol, and 18% were obese. The serum free thyroxine concentrations showed a statistically significant correlation with triglyceride and body mass index, respectively (P<.01), but not with blood pressure, glucose level, or high-density lipoprotein cholesterol level. According to the presence of 0, 1, 2, and 3 or more features of MS, age and sex-adjusted means of serum free thyroxine were 17.8+/-3.7, 17.6+/-3.7, 17.5+/-3.7, and 17.1+/ 3.3 pmol/L, respectively, with a modest, but statistically significant, decreasing trend (P<.05). When comparing subjects in the highest and lowest quartile of free thyroxine, the former group demonstrated a 2-fold decrease in the odds ratio for MS with 3 or more metabolic features. Low circulating free thyroxine levels, albeit normal, were associated with MS in a Chinese population. Further study is necessary to document the role of thyroid hormones in metabolic abnormalities of MS. PMID- 16253644 TI - The influence of sex on the protein anabolic response to insulin. AB - We hypothesize that sex influences whole-body protein anabolism in the postabsorptive state and in response to hyperinsulinemia. Kinetics of 3-(3)H glucose and (13)C-leucine were studied in 16 men and 15 women after energy- and protein-controlled diets, before and during a hyperinsulinemic, euglycemic, isoaminoacidemic clamp. In the postabsorptive state, women had 20% higher rates of leucine Ra (protein breakdown) and nonoxidative Rd (synthesis) adjusted for fat-free mass than men but net leucine balance was as negative. In response to hyperinsulinemia, leucine oxidation rates increased only in women and the change in net leucine balance was less than in men. Net leucine balance during the clamp correlated with rates of glucose disposal. Thus, women showed greater protein turnover rates when adjusted for fat free mass in the postabsorptive state, and lesser insulin sensitivity of protein anabolism and net protein accretion. A relationship exists between the protein anabolic response to insulin and the insulin sensitivity of glucose metabolism. PMID- 16253645 TI - Puerariae radix prevents bone loss in castrated male mice. AB - Puerariae radix (PR) is one of the earliest and most important crude herbs used in Chinese medicine for various medicinal purposes. PR contains a high amount of isoflavonoids, such as daidzein and genistein, which are known to prevent bone loss induced by estrogen deficiency. We have demonstrated that PR not only completely prevents bone loss but also significantly increases the bone mass at high doses in ovariectomized mice without exhibiting estrogenic action in the uterus. In this study, we examined whether PR exhibits effects on bone loss in androgen-deficient male mice similar to estrogen-deficient female mice. Male mice were orchidectomized (ORX) and fed a diet containing low, middle, and high doses (5%, 10%, and 20% of diet, respectively) of PR or normal diet with subcutaneous administration of 17beta-estradiol (E(2), 0.03 microg/d; Sigma, St Louis, Mo), for 4 weeks. In ORX mice, the seminal vesicle weight decreased markedly, and it was not affected by the administration of any doses of PR and E(2). The bone mineral density (BMD) of the whole femur was significantly decreased by ORX, and the decrease in BMD was completely prevented by intake of the diet with the low dose of PR. Intake of the diet with the middle dose of PR further normalized BMD in ORX mice. Furthermore, the high dose of PR administration (PR20) significantly increased BMD in ORX mice, and the potency was similar to that of E(2). Morphometric analysis of the femoral metaphysis showed that intake of the diet with the low dose of PR completely prevented the decrease in bone volume/tissue volume and trabecular number and restored the increase in trabecular separation in ORX mice. In addition, intake of the diet with the high dose of PR further increased bone volume/tissue volume and trabecular number and decreased trabecular separation in ORX mice. These results propose the possibility that estrogenic Chinese herbs such as PR can be one of the candidates for the treatment or prevention of osteoporosis in elderly men with hypogonadism. PMID- 16253646 TI - Ketone body metabolism in lean and obese women. AB - Previous studies have demonstrated decreases in whole-body and muscle fat oxidation in obese individuals. Because muscle also uses ketone bodies, and because the ketone body oxidation pathway differs from that of fatty acid oxidation, this study was initiated to determine whether there were differences in ketone body metabolism between obese and lean subjects. Plasma beta-hydroxy butyrate (beta-OHB) concentration was measured in 47 lean and 47 age-matched obese women, and the rate of beta-OHB oxidation by muscle homogenates was measured in a subset of 8 lean and 8 obese women. Plasma free fatty acid levels, which have been reported to correlate with ketone body production, were higher (P<.05) in the obese than in the lean women (662+/- 46 and 463+/- 44 nmol/L, respectively) as was plasma insulin level. However, the beta-OHB concentration was lower in obese than in lean subjects (235+/-17 and 323+/-29 micromol/L, respectively; P<.05). The rate of beta-OHB oxidation was also lower (P<.05) in muscle of the obese than that of the lean group (139.6+/-12.6 vs 254.6+/-30.0 nmol of CO(2) produced per gram of tissue per hour). These data illustrate that production and use of ketone bodies are lower in obese women than in lean controls. The decreased oxidation of ketone bodies by muscle is consistent with aberrations in muscle metabolism in the obese individuals that most likely relates to a decrease in mitochondrial numbers. PMID- 16253647 TI - Transforming growth factor beta1 release by human adipose tissue is enhanced in obesity. AB - The present studies examined the effect of obesity in humans on the release of transforming growth factor beta1 (TGF-beta1) by human adipose tissue. The regulation of TGF-beta1 release by adipose tissue as well as the question of whether its release is due to the adipocytes or the nonfat cells in adipose tissue was also examined. There was a statistically significant (r=0.50) correlation between the body mass index of the fat donors and the subsequent release of TGF-beta1 release by subcutaneous adipose tissue. There was also a positive correlation between total TGF-beta1 release by adipose tissue explants and body fat content (r=0.69). The question of whether tumor necrosis factor alpha (TNF-alpha) and/or interleukin 1 beta (IL-1 beta) regulate the release of TGF-beta1 was investigated by incubation of adipose tissue explants with a soluble human TNF-alpha receptor (etanercept) and a neutralizing antihuman IL-1 beta antibody. The release of TGF-beta1 over 48 hours by adipose tissue explants was significantly enhanced in the presence of both the inhibitor of TNF-alpha and of IL-1 beta. It is of interest, in view of the elevated circulating insulin in blood of morbidly obese women, that the release of TGF-beta1 by adipose tissue was enhanced in the presence of insulin. The question of whether the release of TGF-beta1 by human adipose tissue explants was primarily due to adipocytes, as is the case for leptin, or the nonfat cells present in human adipose tissue, as is the case for IL-8 and prostaglandin E(2), was examined. The release of TGF-beta1 was primarily by the nonfat cells of human adipose tissue because release by adipocytes was less than 10% of that by the nonfat cells of adipose tissue. PMID- 16253648 TI - The C161-->T polymorphism in peroxisome proliferator-activated receptor gamma, but not P12A, is associated with insulin resistance in Hispanic and non-Hispanic white women: evidence for another functional variant in peroxisome proliferator activated receptor gamma. AB - The P12A variant in the peroxisome proliferator-activated receptor gamma (PPARgamma) gene has been intensely studied for association with obesity-related or type-2 diabetes-related traits; however, the results have been somewhat inconsistent in different populations. We genotyped a large cohort of Hispanic and non-Hispanic white individuals from the San Luis Valley Diabetes Study for P12A and another common variant, C161-->T, in the PPARgamma gene to determine if these sites were associated with fasting glucose, insulin, free fatty acid levels, insulin sensitivity, or body fat. There were no statistically significant frequency differences at these two sites between Hispanic and non-Hispanic individuals. No significant association with the metabolic phenotypes was observed for either of the polymorphisms in men; however, in women, significant associations were shown between the C161-->T variant and fasting insulin (P=.008) and the homeostasis model assessment of insulin resistance (HOMA IR; P=.007). After adjusting for age, smoking, fat mass, and skin reflectance, linear regression showed that C161-->T explained 1.5% of the variation in both fasting insulin (P=.031) and HOMA IR (P=.028) whereas P12A contributed only 0.04% (fasting insulin, P=.268) and 0.02% (HOMA IR, P=.418) to the total trait variation. In the San Luis Valley Diabetes Study female patients, C161-->T appears to be a better predictor of fasting insulin levels and insulin resistance than P12A although the effect of this variant is small. These results support the hypothesis that C161-->T is in linkage disequilibrium with unidentified functional variation in PPARgamma or in a linked gene. This could explain some of the inconsistencies in the P12A association studies as the allele frequency and level of linkage disequilibrium of another functional polymorphism in the region could vary in different populations. PMID- 16253649 TI - Asymptomatic atrial fibrillation. AB - Atrial fibrillation (AF) is a significant cause of morbidity and health care expenditures. Patients with AF suffer a variety of symptoms including chest pain, palpitations, shortness of breath, and fatigue. Some patients have no symptoms, a condition referred to as asymptomatic or "silent" AF. Asymptomatic AF has significant clinical implications. Patients with unrecognized AF may present with devastating thromboembolic consequences or a tachycardia-mediated cardiomyopathy. The incidence of asymptomatic AF is greater than previously perceived. This manuscript provides an overview of the clinical entity of asymptomatic AF including the epidemiology, clinical significance, and the implications it has on the daily management of patients suffering from AF. PMID- 16253650 TI - Cardioversion of atrial fibrillation. PMID- 16253652 TI - Rate control vs rhythm control strategies in atrial fibrillation. AB - Rhythm management for atrial fibrillation is a subject of intense interest. In this review, the two main strategies for rhythm management in atrial fibrillation, the "heart rate control" strategy and the "heart rhythm control" strategy, are discussed under several headings. Some of the basic issues surrounding heart rate control and heart rhythm control and their study in clinical trials are reviewed. Seven published trials that deal with a comparison of these two strategies are reviewed in some detail. In summary, no distinct advantage of the rhythm control strategy has been identified so far. With respect to pharmacologic therapies and the type of patient studied in most of these trials, advantages seem to accrue to the rate control strategy. Those advantages include fewer adverse drug effects, fewer hospitalizations, and lower cost. Two trials studying quite different patients, and in one case quite different drug therapies, are currently in progress, and these two trials are also briefly reviewed. Current guidelines have not taken into account fully the findings of the studies described in this review, and comment is offered on the current guidelines. Finally, some of the possibilities for future research around this question are highlighted. PMID- 16253653 TI - Selection of drugs in pursuit of a rhythm control strategy. AB - A rhythm control strategy based on a combination of antiarrhythmic drugs and electrical cardioversion(s) has emerged as a viable alternative for treatment of atrial fibrillation, particularly when the arrhythmia is associated with symptoms, which may be poorly tolerated, or with congestive heart failure. Several classes of antiarrhythmic drugs are available to restore sinus rhythm by chemical cardioversion and to help maintain it once it has been achieved. In our discussion, we will detail aspects of their efficacy and safety, and attempt to outline a pragmatic clinical approach to their use. Some newer drugs are currently under investigation and hold promise for improved efficacy and/or more favorable side effect profile and will be mentioned in this article. PMID- 16253651 TI - Antithrombotic therapy for stroke prevention in atrial fibrillation. AB - Atrial fibrillation (AF) is the most potent common risk factor for ischemic stroke. The number of Americans with nonvalvular AF is expected to increase markedly over the next several decades, making AF-related stroke an important public health concern. Given the individual and societal burden associated with AF-related stroke, efforts to identify and implement efficacious and acceptably safe therapeutic stroke prevention strategies are paramount. This article reviews the existing randomized trial evidence supporting the efficacy of oral vitamin K antagonists (ie, warfarin) or aspirin for preventing thromboembolism in AF, as well as completed and ongoing studies exploring novel antithrombotic agents including the oral direct thrombin inhibitor, ximelagatran, other antiplatelet agents (eg, clopidogrel), factor Xa inhibitors, and other pharmacological agents and additional therapeutic approaches such as mechanical devices and surgical procedures to obliterate the left atrial appendage. PMID- 16253654 TI - Selection of drugs in pursuit of rate control strategy. AB - Atrial fibrillation is the most common sustained arrhythmia. Based on multiple large randomized trials, rate control therapy has been shown to be safe and effective and is gaining greater acceptance as a frontline alternative to drugs to maintain sinus rhythm. Adequate rate control can be achieved by atrioventricular nodal blocking agents both in the acute and chronic settings. In refractory patients, other methods such as atrioventricular node ablation can be used to control rate. PMID- 16253656 TI - CT radiation dose and image quality. AB - Image quality is proportional to radiation dose. Improvements in image quality come at a cost of increased radiation dose. CT scanners are more robust today than even 5 years ago. X-ray tubes are capable of producing high levels of almost continuous radiation for rapid CT volume acquisitions and angiographic studies. All parameters of modern CT scanners provide rapid subsecond, large-volume CT acquisitions. How high we go needs to be tempered by how high we should go regarding radiation dose from CT examinations. Radiologists, referring physicians, medical physicists, CT technologists, CT equipment manufacturers, and regulators need to evaluate the appropriateness of radiation dose for different CT studies. PMID- 16253657 TI - Multidetector CT angiography of the abdomen. AB - Multidetector CT angiography (MDCTA) is redefining traditional imaging strategies of the vascular structures of the abdomen. Angiographic depiction of normal and variant anatomy is becoming the standard for evaluation and has a significant impact in transplant and oncologic surgery. MDCTA is increasingly being used for assessing diseases affecting the vasculature of the abdominal organs, including the abdominal aorta for treatment planning and post therapy follow-up. PMID- 16253658 TI - Liver and biliary system: evaluation by multidetector CT. AB - CT commonly is indicated for the evaluation of suspected hepatic and biliary pathology. The recent introduction of multidetector CT (MDCT) provides unique capabilities that are valuable especially in hepatic volume acquisitions, combining short scan times, narrow collimation, and the ability to obtain multiphase data. These features result in improved lesion detection and characterization. Concomitant advances in computer software programs have made three-dimensional applications practical for a range of hepatic image analyses and displays. This article discusses the specific areas of hepatic and biliary pathology where MDCT has a significant diagnostic impact. PMID- 16253659 TI - Multidetector CT of the pancreas. AB - CT is the primary imaging modality of the pancreas. This article reviews the multidector CT technique and its current status in the diagnosis of various pancreatic diseases. Special emphasis is given to the impact of multidetector CT and postprocessing imaging techniques on the staging of pancreatic adenocarcinoma. PMID- 16253660 TI - Renal multidector row CT. AB - Multidetector row CT is the most recent advance in CT technology. An increased number of detector rows and more powerful x-ray tubes result in faster scanning time, increased volume coverage, and improved spatial and temporal resolution. MDCT technology allows superior image quality, decreased examination time, and the ability to perform complex multiphase vascular and three-dimensional examinations. PMID- 16253661 TI - Multislice CT colonography: current status and limitations. AB - CT colonography (CTC) is a promising method for colorectal cancer screening because it provides a full structural evaluation of the entire colon. It has a superior safety profile, a low rate of complications, and high patient acceptance. In addition, CTC offers the real possibility of eliminating the cathartic bowel preparation, one of the biggest obstacles to patient compliance with colorectal cancer screening. Results of CTC studies in recently published literature are extremely encouraging, demonstrating that this method of screening can detect lesions equal to or larger than 8 mm with few false-positive findings. PMID- 16253662 TI - Multidetector row CT of the small bowel. AB - Multidetector row CT (MDCT) has become an imaging technique of choice to study routinely the small bowel. Thin collimation and fast scanning allow coverage of the entire abdomen within a single suspended respiration phase allowing the use of multiple enhancement phases after intravenous contrast administration. MDCT of the small bowel can identify and stage most of the common diseases of the small bowel. MDCT is changing the paradigm for diagnosing small bowel disease by becoming the first diagnostic line for almost all small bowel diseases. MDCT has the needed sensitivity and specificity, the availability, and the safety for a front-line diagnostic method. PMID- 16253663 TI - Multidetector CT evaluation of abdominal trauma. AB - CT is the imaging modality of choice to evaluate blunt abdominal trauma. With the advent of multidetector CT (MDCT), scanning times have progressively decreased while image resolution has increased owing to thinner collimation and reduced partial volume and motion artifacts. MDCT also allows high quality two dimensional and three-dimensional multiplanar reformatted images to be obtained, which aid in the diagnosis of the complex multisystem injuries seen in the trauma patient. This article describes the authors' current imaging protocol with 16 detector MDCT, the spectrum of CT findings seen in patients with blunt abdominal injuries, and the role MDCT has in guiding injury management. PMID- 16253664 TI - Multidetector CT of the female pelvis. AB - In the emergency room setting, multidetector detector CT (MDCT) offers rapid, noninvasive, multiplanar evaluation of female patients who have acute pelvic pain. MDCT has been integrated into several of the major trauma centers, and its use may surpass the use of ultrasound in the trauma evaluation of the pregnant patient. In the nonemergent setting, MDCT can be used to stage gynecologic malignancy and to evaluate tumor recurrence. Multiplanar MDCT has received some acceptance for evaluation of small primary tumor volume and small metastatic implants. MDCT also has a role in the evaluation of pelvic varices and suspected pelvic congestion syndrome. PMID- 16253665 TI - Angiographic imaging of the lower extremities with multidetector CT. AB - Multidetector CT (MDCT) has improved imaging of the arteries in the lower extremities. The main advantages of this novel technology are the exceptionally fast scan times, high spatial resolution, increased anatomic coverage, and capability to generate high-quality multiplanar reformations and three dimensional (3-D) renderings from raw data that can be reprocessed easily and quickly. The applications of MDCT in imaging the lower extremities are multiple and varied. They include the evaluation of peripheral arterial occlusive and aneurysmal disease, the patency and integrity of bypass grafts, and arterial injury owing to trauma. This article describes the techniques of lower extremity MDCT angiography and its use in a few clinical applications. PMID- 16253669 TI - The impact of subspecialization on postgraduate medical education in neurosurgery. AB - Medical subspecialization is a response to rapidly expanding technology and knowledge. Although beneficial to patient care, it poses a challenge to the current infrastructure of resident education. This article analyzes the advent of subspecialization, the current template of postgraduate neurosurgical education, the impact of subspecialization on postgraduate neurosurgical education, and, finally, suggests strategies to optimize professional education in the face of an increasingly subspecialized field. PMID- 16253666 TI - Tiamulin resistance in porcine Brachyspira pilosicoli isolates. AB - There are few studies on antimicrobial susceptibility of Brachyspira pilosicoli, therefore this study was performed to investigate the situation among isolates from pigs. The tiamulin and tylosin susceptibility was determined by broth dilution for 93 and 86 porcine B. pilosicoli isolates, respectively. The isolates came from clinical samples taken in Swedish pig herds during the years 2002 and 2003. The tylosin minimal inhibitory concentration (MIC) was >16 microg/ml for 50% (n=43) of the isolates tested. A tiamulin MIC >2 microg/ml was obtained for 14% (n=13) of the isolates and these were also tested against doxycycline, salinomycin, valnemulin, lincomycin and aivlosin. For these isolates the susceptibility to salinomycin and doxycycline was high but the MICs for aivlosin varied. The relationship between the 13 tiamulin resistant isolates was analyzed by pulsed-field gel electrophoresis (PFGE). Among the 13 isolates 10 different PFGE patterns were identified. PMID- 16253680 TI - Radiosurgery for dural arteriovenous fistulas. AB - OBJECTIVE: Dural arteriovenous fistulas (DAVFs) comprise 10% to 15% of all arteriovenous malformations. Recent studies have demonstrated promising results when radiosurgery is used for DAVFs. We retrospectively analyzed our patients with DAVFs who received stereotactic radiosurgery with or without embolization. METHODS: Between 1991 and 2002, 18 patients with 23 angiographically confirmed symptomatic DAVFs underwent gamma knife radiosurgery, either alone (n = 8) or in combination with embolization (n = 10). A retrospective chart review was performed to identify DAVF location, venous drainage pattern, radiosurgery dosimetry, clinical outcomes, and imaging results. The series included 9 men and 9 women with a mean age of 65 (range 50-89) years. Nine patients received particulate, coil, and/or absolute ethanol embolization before radiosurgery, and 1 patient received particulate embolization after radiosurgery. The mean duration of clinical follow-up was 43 (range 2-116) months. The mean margin radiosurgery dose was 20 (range 15-30) Gy. RESULTS: Nine patients had complete resolution of their presenting symptoms, and 9 patients had resolution of all but 1 of their presenting symptoms. Angiographic follow-up (mean 46 months) was performed on 8 patients demonstrating complete obliteration in all the cases. Seven patients evaluated by magnetic resonance angiography or computed tomography angiography showed no evidence of DAVF (4 patients) or decreased DAVF size (3 patients). After radiosurgery, 1 patient developed a temporary hemiparesis. Two permanent neurological deficits occurred after embolization before radiosurgery. No patient had an intracranial hemorrhage after treatment. CONCLUSION: Stereotactic radiosurgery provides effective long-term relief of symptoms in selected patients with DAVFs. PMID- 16253683 TI - Primary central nervous system lymphoma of the brain stem responding favorably to radiosurgery: a case report and literature review. AB - OBJECTIVE: To explore various therapeutic alternatives and evaluate the clinical results of patients with primary central nervous system lymphoma (PCNSL). We report a case of PCNSL with involvement of the brain stem managed with radiosurgery using the Leksell gamma knife as the treatment modality. CASE DESCRIPTION: A 55-year-old white woman presented with complaints of weakness on the left side of her body and double vision associated with a frontal headache. Nervous system examination revealed right-sided oculomotor palsy and left-sided motor paresis, grade 2/5, in both upper and lower limbs. Her Karnofsky performance score was 50. A contrast-enhanced magnetic resonance imaging scan of the brain revealed an irregular space-occupying lesion in the pons that enhanced uniformly with gadolinium. A frame-based stereotactic biopsy was performed revealing a B-cell malignant lymphoma. Subsequently, stereotactic radiosurgery using the Leksell gamma knife was performed. A dose of 11 Gy was delivered to the tumor margin. A follow-up magnetic resonance imaging scan after 2 months revealed a 50% reduction in the size of the enhancing lesion. CONCLUSION: Our own results and limited evidence from the literature suggest stereotactic radiosurgery as a potentially safe and effective treatment option in patients with PCNSL. PMID- 16253685 TI - Gamma knife radiosurgery for cavernous sinus metastases and invasion. AB - BACKGROUND: We evaluated the efficacy of gamma knife radiosurgery for cavernous sinus metastases and invasion. METHOD: We treated and followed up 21 patients with cavernous sinus metastases and invasion using gamma knife radiosurgery. Nine of these patients had nasopharyngeal cancer, and 12 had distant metastases from other cancers. The volume of tumors ranged from 2.9 to 50.0 (median 9.9) mL. and the radiation dose to the tumor margin was 10 to 21 (median 14) Gy. RESULTS: The median follow-up period was 9 months. Clinical symptoms were improved in 48% of the patients after treatment, and tumor growth control was obtained in 67% of the patients at their final follow-up. The actual 1- and 2-year tumor growth control rates were 68% and 43%, respectively. The mean survival time was 13 months. No patient had radiation injury. CONCLUSION: Gamma knife radiosurgery is a very useful therapeutic option for the treatment of cavernous sinus metastases and invasion, either as initial treatment or as an adjunct treatment for recurrences even in preirradiated patients. PMID- 16253687 TI - Application of neuronavigator coupled with an operative microscope and electrocorticography in epilepsy surgery. AB - Application of neuronavigator coupled with an operative microscope and electrocorticography (ECoG) is a new trial for epilepsy surgery for achieving better seizure outcome and better efficiency for lesionectomy. We used a neuronavigator coupled with a microscope to delineate the magnetic resonance image-detected lesion for lesionectomy and used ECoG for evaluation of the epileptogenic foci. There were 46 patients with medically intractable partial seizures who underwent craniotomy for epilepsy surgery. Half of the patients had lesions at the temporal lobe and another half at the extratemporal lobe. Sixty one percent of the patients were seizure-free (grade I) and 22% were nearly seizure-free (grade II). Overall, 83% of the patients had satisfactory seizure control. Complete lesion removal was successful in 37 patients (80%). For lesions with requiring complete removal, neuronavigator coupled with a microscope was 95% effective for lesionectomy. Class A of postresection ECoG had a higher rate of seizure-free outcome (92%) (P < .05). On the other hand, 93% of patients (26/28) with seizure-free outcome (grade I) needed complete lesion resection (P < .05). Lesions at the extratemporal lobe yielded a higher rate of seizure-free outcome (78.2%, 18/23) (P < .05). Application of neuronavigator and ECoG (additional cortical resection) is usually necessary for temporal lobe lesions. All patients with cavernous hemangioma were seizure-free. The complication rate in our study was 8.7%. Neuronavigator coupled with a microscope provides efficacy and safety to complete lesionectomy, which is a key point of seizure outcome. Intraoperative ECoG is valuable for evaluating the epileptogenic foci for epilepsy surgery especially for lesions at the temporal lobe. PMID- 16253690 TI - Frequent association of cortical dysplasia in dysembryoplastic neuroepithelial tumor treated by epilepsy surgery. AB - BACKGROUND: We report on our experience with epilepsy surgery in the treatment of localization-related epilepsy caused by dysembryoplastic neuroepithelial tumor (DNT) aimed at achieving the best seizure control. METHODS: A retrospective analysis was performed on the pathological reports as well as on clinical data of 24 case patients with medically intractable epilepsy with DNT treated surgically between 1995 and 2000 at the Samsung Medical Center. Resective surgery was performed using subdural electrodes or intraoperative electrocorticography in all patients. RESULTS: The mean follow-up period was 57.2 months. Two patients had rare seizures transiently after surgery but remained free from seizures after 6 months. Others remained completely free from seizures. There was a strong tendency of temporal lobe involvement (19 cases; 79.2%). Size of tumors located at medial temporal regions was significantly smaller than those at lateral temporal or frontal lobes (P < .05). A rather radical resection (tumor plus surrounding tissue showing active epileptogenicity) was performed in all but one case where only focal lesionectomy was done. In 20 of the 24 cases (83.3%), association of cortical dysplasia (CD) was found on pathological examination. CONCLUSIONS: We conclude that DNT is frequently associated with CD, with a wide area of epileptogenic activity that might be related to the presence of CD around the DNT. Comprehensive preoperative investigations for accurate localization of epileptogenic activity, meticulous brain mapping, and a rather radical resection of pathological areas might be essential for the achievement of excellent seizure control in DNT-associated epilepsy. PMID- 16253691 TI - Epidermoid cysts of the cavernous sinus. AB - BACKGROUND: Epidermoid cysts involving the cavernous sinus are rare lesions. They can be divided into 3 different categories: extracavernous, interdural, and true intracavernous. METHODS: This classification of cavernous epidermoid cysts is correlated with the extent of tumor resection and the clinical outcome in reported cases here and elsewhere. RESULTS: Patients with cysts invading or compressing the cavernous sinus present with an excellent functional recovery after surgery independent of the extent of tumor resection. In cases of interdural cysts, there is often an adherence of the tumor capsule to the third and fourth cranial nerves. Therefore, in this group, subtotal resection is a safe strategy to prevent surgical morbidity. However, this policy may lead to a higher incidence of tumor recurrence in these cases. In true intracavernous lesions, a higher rate of total removal is possible despite the increased occurrence of internal carotid artery encasement and cranial nerve displacement. CONCLUSION: Postoperative outcome with preservation of neural and vascular structures in cavernous epidermoid cysts is more likely related to surgical strategy than to tumor localization. Independent of the different tumor origins in these cases, a subtotal resection is usually sufficient to achieve postoperative improvement of the symptoms without additional morbidity. PMID- 16253694 TI - Plasma cell tumors of the skull. AB - BACKGROUND: Plasma cell tumors are rare neoplasms, which include extramedullary plasmacytoma, solitary plasmacytoma of the bone, and multiple myeloma. Although indistinguishable histopathologically, these entities need to be differentiated as treatment and prognosis vary. METHODS: This study was conducted by retrospective chart review and correspondence with patients diagnosed to have plasma cell tumors of the skull from 1992 to 2004. The clinico-diagnostic investigations, treatment, and subsequent follow-up of the patients were obtained. RESULTS: We report 5 cases of plasma cell tumors of the skull. The lesion involved the base of skull in 2 and the calvarium in 3 patients. In 2 cases, the histopathology was reported as plasmacytoma and further investigations revealed systemic involvement, indicative of multiple myeloma. In 1 patient, the biopsy was reported as multiple myeloma. The above 3 patients underwent radiation, followed by chemotherapy, and there was improvement in their neurologic status at subsequent follow-up. Another patient with a scalp swelling and symptoms of systemic involvement underwent investigations to detect multiple myeloma and, hence, was not subjected to an invasive procedure for histopathologic diagnosis. Chemotherapy was initiated but was discontinued because of social reasons. The last patient in our series died in the immediate postoperative period and although her histopathology was reported as plasmacytoma, she could not be evaluated for systemic involvement. CONCLUSION: The treatment options are different for the systemic form of disease and the localized disease; hence, it is imperative that a rigorous search for systemic involvement is conducted in a case of solitary or extramedullary plasmacytoma. PMID- 16253696 TI - Percutaneous coagulation of choroid plexus to unblock the ventricular catheter using the Seldinger technique: preliminary report. AB - BACKGROUND: Blockage of the ventricular catheter is the most common reason for shunt malfunction and is usually due to ingrowth of choroid plexus into the catheter lumen. The authors describe a simple technique to unblock the ventricular catheter in situ. METHODS: In children with symptoms and radiological evidence of shunt malfunction, the Rickham reservoir was tapped under a general anesthetic. Manometry with saline was performed to confirm isolated proximal blockage of shunt. A guide wire, insulated to the scalp, was passed into the ventricular catheter using the Seldinger technique. Bursts of monopolar diathermy were applied to the guide wire to coagulate choroids plexus and manometry was repeated. In this preliminary study, all ventricular catheters were subsequently revised. RESULTS: This percutaneous technique was undertaken in 7 children with confirmed proximal shunt malfunction. Successful unblocking of the ventricular catheter was established by increased flow of cerebrospinal fluid around the guide wire. The mean +/- SEM manometry reading post-diathermy (21.9 +/- 5 cm) was higher than the pre-diathermy value (0.6 +/- 0.6 cm; P < .01). CONCLUSIONS: Percutaneous coagulation of the choroids plexus using the Seldinger technique is a simple, inexpensive, minimally invasive method to unblock ventricular catheters and it may be suitable as a stand-alone technique in selected patients. PMID- 16253697 TI - Bilateral chronic subdural hematoma cases showing rapid and progressive aggravation. AB - BACKGROUND: We have analyzed the records of our own hospitalized cases of bilateral chronic subdural hematoma (cSDH) to reveal the prognosis. METHODS: Ninety-eight cases of cSDH were operated at our hospital over a 6-year period, in which 14 cases were classified as being bilateral. Among these 14 cases, 6 cases showed a rapid and aggressive clinical course. Therefore, complicated risk factors, the initial data on coagulofibrinolytic examination, magnetic resonance imaging appearance, and prognosis were analyzed. RESULTS: Of the 6 cases, 5 showed a rapid aggravation as they awaited surgery. The period of the aggravation since the initial diagnosis harboring cSDH was 19 to 54 hours. One case was at first neurologically free from any disturbance but 17 hours later experienced a generalized seizure. All 6 cases experienced consciousness disturbance. In addition, 3 of them manifested oculomotor palsy. Two cases showed an abnormality of coagulofibrinolytic activity. No significant risk factors were revealed. In 4 cases, T(2)-weighted images (T2WIs) revealed the hematoma of a mixed high and low intensity, indicating that the hematoma consisted of both liquid and solid parts of a freshly formed blood clot. In 2 cases, the hematoma showed a low intensity in T(1)-weighted image (T1WI), indicating a recent bleeding of a significant amount. CONCLUSION: The bilateral cases of cSDH should be treated as early as possible with simultaneous decompression of bilateral hematoma pressure, even if the patient shows minimal neurologic deficits. Mixed high and low intensity in T2WI or low intensity in T1WI is the most predictable factor to show rapid aggravation. PMID- 16253699 TI - Urgent treatment of severe subarachnoid hemorrhage caused by ruptured traumatic aneurysm of the cavernous internal carotid artery using coil embolization followed by superficial temporal artery-middle cerebral artery anastomosis: a case report. AB - BACKGROUND: Traumatic aneurysm of the cavernous internal carotid artery (ICA) with extension into the subarachnoid space is associated with increased risk of fatality especially when it is accompanied by severe subarachnoid hemorrhage (SAH). Only cases of patients who survived the acute stage and who were treated in a delayed setting have been reported. There has been no successfully treated case immediately after an injury. CASE DESCRIPTION: We encountered a 48-year-old man who presented with dense SAH immediately after being involved in a motor vehicle accident. Emergent angiography revealed traumatic aneurysm of the left cavernous ICA with extension beyond the superior wall of the cavernous sinus into the subarachnoid space and concomitant direct high-flow carotid cavernous fistula. Detachable platinum coil occlusion of the cavernous ICA followed by superficial temporal artery-middle cerebral artery anastomosis on day 0 and aggressive therapy to SAH, including ventriculocisternal irrigation and drainage, was performed. The patient eventually made a good recovery. CONCLUSION: Considering the extremely poor prognosis and unstable nature of a ruptured traumatic aneurysm with extensive SAH in the acute stage, definitive and immediate prevention of rebleeding in conjunction with proper revascularization would be warranted, such as in the present case. PMID- 16253701 TI - Staged bilateral vertebral artery occlusion for ruptured dissecting aneurysms of the basilar artery: a report of 2 cases. AB - BACKGROUND: Dissecting aneurysm of the basillary artery BA is rare. Although mortality rate is high, management remains controversial. We report 2 cases of dissecting aneurysm of the BA presenting with subarachnoid hemorrhage (SAH), both of which were successfully treated using staged occlusion of bilateral vertebral arteries (VAs). CASE DESCRIPTION: A 64-year-old man and a 34-year-old woman presented with SAH associated with ruptured dissecting aneurysm of the BA. After endovascular occlusion of a single VA, blood flow in the dissected lumen was reduced. However, one aneurysm rebled and the bleb of the other did not change. Vertebral arteries were also occluded using endovascular techniques at 4 and 2 weeks after initial treatment, respectively. On the second intervention, stump pressure ratios of VAs intended for occlusion were 62.5% and 50.6%, respectively. The patients tolerated temporary occlusion of bilateral VAs well. Subsequent permanent occlusion of bilateral VAs resulted in no neurological complication. Complete obliteration of the aneurysmal lumen was demonstrated on magnetic resonance angiography performed 72 and 5 months later, respectively. CONCLUSION: Staged bilateral VA occlusion might be the last recourse to prevent further hemorrhage from BA dissecting aneurysm. The technique can be safely applied when the stump pressure ratio is 50.6% or greater and when the patient tolerates temporary occlusion, which suggests the existence of sufficient collateral flow from the anterior circulation. PMID- 16253705 TI - Spinal aneurysm arising from the feeding pedicle of a thoracic perimedullary arteriovenous fistula: case report. AB - BACKGROUND: Aneurysms of the spinal artery are exceedingly rare. We report on the first case of a pathologically proven aneurysm arising from the feeding pedicle of a perimedullary arteriovenous fistula (AVF). CASE DESCRIPTION: A 42-year-old man presented with subarachnoid hemorrhage (SAH) and spinal angiography demonstrated a thoracic perimedullary AVF fed by the left T8 intercostal artery that also harbored a spinal aneurysm. He underwent laminectomy 18 days postonset. The aneurysm was resected and the fistula was interrupted. CONCLUSION: For appropriate treatment planning, it is necessary to rule out the presence of aneurysms on the feeding vessel in patients with spinal perimedullary AVF who present with SAH. PMID- 16253703 TI - Clinical presentation and surgical management of dissecting posterior inferior cerebellar artery aneurysms: 2 case reports. AB - Intracranial dissection presenting with subarachnoid hemorrhage (SAH) most commonly involves the vertebral artery. The natural history of this lesion suggests frequent early rehemorrhage and need for urgent treatment. Isolated dissection of the posterior inferior cerebellar artery (PICA) is very rare. We present 2 cases of isolated PICA dissections presenting with SAH. Both patients were middle-aged men who presented with transient loss of consciousness, severe headache, and cranial neuropathies. Initial angiography showed dilatation and narrowing of PICA consistent with dissection and aneurysm formation. The vertebral arteries were normal and there was no other cause for the SAH. Repeat angiography 2 weeks after admission revealed significant enlargement of the aneurysmal dilation of the dissected segment of PICA in both patients. Both patients were treated operatively. One patient had clip reconstruction of the PICA with preservation of flow through the PICA. Follow-up angiography one year later showed no recurrence. The other patient underwent direct surgical trapping and resection of the dissected segment of PICA after passing balloon occlusion testing at the vertebral-PICA junction. Both patients have more than 2 years of clinical follow-up and remain well. Isolated PICA dissection seems to have a less ominous natural history compared to vertebral artery dissection. These lesions need to be followed carefully for evidence of aneurysmal enlargement. Direct surgical reconstruction of the dissected segment may be possible. Balloon occlusion testing may be very helpful in determining if the involved PICA segment can be sacrificed. PMID- 16253706 TI - Is there a natural right to medical care? PMID- 16253708 TI - Re: The controversial arcade of Struthers. PMID- 16253709 TI - Report on the surgical clinic at the Hotel-Dieu. 1831. AB - Concerning finger contracture as a result of a condition affecting the palmar fascia. A description of the illness. A surgical operation that is suitable for it. PMID- 16253710 TI - Letters to the editor. PMID- 16253713 TI - Secondary renal amyloidosis due to long-standing tubulointerstitial nephritis in a patient with Sjogren syndrome. AB - A 53-year-old patient with long-standing primary Sjogren syndrome presented with acute renal failure and nephrotic syndrome caused by secondary (AA) renal amyloidosis. Ten years before, he had been admitted because of exacerbation of the systemic disease. At that time, a pseudolymphoma of the kidney was diagnosed. To our knowledge, this is the first report of a patient with primary Sjogren syndrome and secondary (AA) amyloidosis with amyloid deposition in the kidneys causing nephrotic syndrome. PMID- 16253714 TI - Darbepoetin alfa treatment for post-renal transplantation anemia during pregnancy. AB - Anemia is common in patients with chronic kidney disease (CKD) and those who have received a kidney allograft. Anemia is most prevalent in kidney transplant recipients before and immediately after transplantation, but also can occur months after transplantation if the donor kidney begins to fail. Replacement therapy for CKD-related and posttransplantation anemia is effective through the administration of exogenous erythropoiesis-stimulating proteins. Darbepoetin alfa (Aranesp; Amgen Inc, Thousand Oaks, CA) is a unique erythropoiesis-stimulating protein that can be administered at an extended dosing interval relative to recombinant human erythropoietin because of its approximately 3-fold longer serum half-life. Although darbepoetin alfa has been shown to be an effective treatment for patients with anemia of CKD and anemia after kidney transplantation, limited data have been published showing efficacy in treating women with anemia of these conditions during pregnancy. We report a case of successful darbepoetin alfa treatment for severe anemia in a pregnant transplant recipient. PMID- 16253715 TI - Vibrio vulnificus peritonitis after handling of seafood in a patient receiving CAPD. AB - Vibrio vulnificus is a marine bacterium and opportunistic human pathogen. Associated infections have contributed to the majority of seafood-related deaths in the United States. In patients with such predisposed clinical conditions as chronic liver disease, immunocompromised state, and end-stage renal disease, this organism has been associated with the development of life-threatening primary septicemia and severe wound infection. However, continuous ambulatory peritonitis dialysis (CAPD)-related peritonitis caused by V vulnificus has not been reported. We describe a patient receiving CAPD who developed peritonitis caused by V vulnificus after handling seafood. This case highlights the importance of strict aseptic technique during CAPD exchanges and calls for an effort in educating our dialysis patients on precautions about seafood handling. PMID- 16253716 TI - Sudden late onset of gross hematuria in a previous renal transplant recipient 3 months after transplant nephrectomy. AB - Causes of gross hematuria in a patient with end-stage renal disease are limited compared with those in patients with normal renal function. Given the increased likelihood of patients with end-stage renal disease developing renal cell carcinoma, the workup focuses on a careful evaluation of the collecting system. The workup for gross hematuria in a renal transplant recipient is similar; however, the focus shifts toward a more thorough evaluation of the transplanted kidney and bladder because immunosuppression increases the overall risk for malignancy. An immunosuppressed patient also is at risk for infectious processes in the transplanted kidney manifesting as gross hematuria. Concerns for chronic rejection also should be investigated, although microscopic hematuria is more common in this scenario. If this is unrevealing, then close scrutiny of the native kidneys for possible sources of bleeding is warranted. We present an interesting and unusual cause of painless gross hematuria in a patient with end stage renal disease and transplant nephrectomy 3 months before the onset of bleeding. PMID- 16253717 TI - Efficiency of the Genius batch hemodialysis system with low serum solute concentrations: the case of lithium intoxication therapy. AB - BACKGROUND: The Genius batch system consists of a 90-L closed reservoir, from which fresh dialysate is extracted at the top and to which spent dialysate is returned at the bottom. It was shown in long-term hemodialysis patients that almost the entire amount of unspent dialysate can be used before contamination of fresh with spent dialysate occurs. Separation is caused by differences in density, partly because of the presence of uremic solutes in spent dialysate. The question is raised whether this separation can be maintained during dialysis of patients who experience an intoxication without renal failure. METHODS: A patient intoxicated with lithium was dialyzed using the Genius system, prepared at 37 degrees C, during 300 minutes. With dialysate flow set at 300 mL/min (5 mL/s) and in the absence of mixing, urea is not expected at the inlet dialysate tubing before minute 300. RESULTS: In the dialysate inlet tubing, an abrupt increase in lithium and urea concentrations was observed 210 minutes after the start of the session, reflecting contamination of fresh with spent dialysate. At minute 210, only 60.9 L of 90 L of dialysate had crossed the dialyzer. In a control dialysis treatment in a patient with marked renal failure, this mixing occurred only at 300 minutes. CONCLUSION: In the present observation, it is shown that during Genius dialysis in a patient without renal failure, an earlier contamination of fresh with spent dialysate can occur, compared to conditions of renal failure. PMID- 16253718 TI - Use of EPO in critically ill patients with acute renal failure requiring renal replacement therapy. AB - BACKGROUND: Recombinant human erythropoietin (EPO) is used widely to treat anemia in patients with chronic kidney disease, but the benefits of EPO use in patients with acute renal failure (ARF) are unclear. In vitro and animal studies suggest that EPO may promote renal recovery and decrease mortality in ARF. METHODS: We conducted a retrospective cohort study at a tertiary-care center to evaluate the use of EPO in 187 critically ill patients with ARF requiring renal replacement therapy. RESULTS: Compared with patients not administered EPO (n = 116), patients administered EPO (n = 71) were significantly more likely to have baseline chronic kidney disease, have undergone vascular surgery, and have received intermittent hemodialysis, rather than continuous renal replacement therapy. In a propensity adjusted analysis that controlled for differences between the 2 cohorts and baseline hemoglobin level, EPO use did not decrease the transfusion of packed red blood cells. Renal recovery was not more common in patients administered EPO: the odds ratio for renal recovery in the propensity-adjusted analysis was 0.63 (95% confidence interval, 0.30 to 1.3) with EPO use. In-hospital survival was more common in the EPO-treated group, but this potential benefit was not significant in propensity-adjusted analyses. CONCLUSION: Although EPO use was not associated with a decrease in transfusion requirements or with renal recovery in our retrospective study, 37% of critically ill patients with ARF were treated with EPO at varying doses. A randomized controlled trial is needed to evaluate the potential benefits of EPO use in patients with ARF. PMID- 16253719 TI - Canadian randomized trial of hemoglobin maintenance to prevent or delay left ventricular mass growth in patients with CKD. AB - BACKGROUND: This randomized clinical trial is designed to assess whether the prevention and/or correction of anemia, by immediate versus delayed treatment with erythropoietin alfa in patients with chronic kidney disease, would delay left ventricular (LV) growth. Study design and sample size calculations were based on previously published Canadian data. METHODS: One hundred seventy-two patients were randomly assigned. The treatment group received therapy with erythropoietin alfa subcutaneously to maintain or achieve hemoglobin (Hgb) level targets of 12.0 to 14.0 g/dL (120 to 140 g/L). The control/delayed treatment group had Hgb levels of 9.0 +/- 0.5 g/dL (90 +/- 5 g/L) before therapy was started: target level was 9.0 to 10.5 g/dL (90 to 105 g/L). Optimal blood pressure and parathyroid hormone, calcium, and phosphate level targets were prescribed; all patients were iron replete. The primary end point is LV growth at 24 months. RESULTS: One hundred fifty-two patients were eligible for the intention-to-treat analysis: mean age was 57 years, 30% were women, 38% had diabetes, and median glomerular filtration rate was 29 mL/min (0.48 mL/s; range, 12 to 55 mL/min [0.20 to 0.92 mL/s]). Blood pressure and angiotensin-converting enzyme inhibitor/angiotensin receptor blocker use were similar in the control/delayed treatment and treatment groups at baseline. Erythropoietin therapy was administered to 77 of 78 patients in the treatment group, with a median final dose of 2,000 IU/wk. Sixteen patients in the control/delayed treatment group were administered erythropoietin at a median final dose of 3,000 IU/wk. There was no statistically significant difference between groups for the primary outcome of mean change in LV mass index (LVMI) from baseline to 24 months, which was 5.21 +/- 30.3 g/m2 in the control/delayed treatment group versus 0.37 +/- 25.0 g/m2 in the treatment group. Absolute mean difference between groups was 4.85 g/m2 (95% confidence interval, -4.0 to 13.7; P = 0.28). Mean Hgb level was greater in the treatment group throughout the study and at study end was 12.75 g/dL (127.5 g/L in treatment group versus 11.46 g/dL [114.6 g/L] in control/delayed treatment group; P = 0.0001). LV growth occurred in 20.1% in the treatment group versus 31% in the control/delayed treatment group (P = 0.136). In patients with a stable Hgb level, mean LVMI did not change (-0.25 +/- 26.7 g/m2), but it increased in those with decreasing Hgb levels (19.3 +/- 28.2 g/m2; P = 0.002). CONCLUSION: This trial describes disparity between observational and randomized controlled trial data: observed and randomly assigned Hgb level and LVMI are not linked; thus, there is strong evidence that the association between Hgb level and LVMI likely is not causal. Large randomized controlled trials with unselected patients, using morbidity and mortality as outcomes, are needed. PMID- 16253720 TI - The effect of cholesterol reduction with cholestyramine on renal function. AB - BACKGROUND: Epidemiological studies suggest that dyslipidemia is an independent risk factor for the development of renal insufficiency in otherwise healthy individuals. Yet, data on the effect of cholesterol reduction are lacking in this population. We performed a secondary analysis of a large existing cohort to determine whether treatment with cholestyramine improved renal function compared with placebo. METHODS: A total of 3,603 middle-aged men from the Lipid Research Clinics Coronary Primary Prevention Trial comprised the study group: 1,806 men were randomly assigned to treatment with cholestyramine, and 1,797 men, placebo. The primary outcome is difference in glomerular filtration rates between the 2 groups. RESULTS: A total of 1,806 men were randomly assigned to treatment with cholestyramine, and 1,797 men, placebo. For the entire group, the estimated mean difference in glomerular filtration rates between the cholestyramine and placebo groups was 0.39 mL/min/1.73 m2 (0.007 mL/s/1.73 m2; P = 0.28) during a follow-up period of more than 8 years. CONCLUSION: Cholesterol reduction with cholestyramine treatment did not meaningfully affect renal function compared with placebo in the present analysis. Prospective intervention trials are needed to determine whether decreasing serum cholesterol levels benefits kidney function in otherwise healthy individuals. PMID- 16253721 TI - Comparison and interpretation of urinalysis performed by a nephrologist versus a hospital-based clinical laboratory. AB - BACKGROUND: Urinalysis (UA) is considered the most important laboratory test in evaluating patients with kidney disease. Anecdotally, we have observed differences between results of UA performed by nephrologists compared with those performed by certified medical technologists or clinical laboratory scientists that could affect a clinician's diagnosis. Whether there are differences between UA performed by the clinical laboratory and that performed by a nephrologist was determined, and accuracy of diagnosis based on interpretation of the UA was compared. METHODS: Urine samples were obtained from 26 patients with acute renal failure (ARF). An aliquot of urine was sent to the clinical laboratory for UA. Nephrologist A, blinded to the patient's clinical information, performed a UA on the other aliquot of urine, generated a report, and assigned the most likely diagnosis for ARF based on UA findings. Nephrologist B, also blinded to the clinical information, reviewed nephrologist A's UA reports and assigned a diagnosis for ARF to each report. Nephrologists A and B both assigned a diagnosis (or diagnoses) for the ARF based on laboratory UA results. These 4 sets of diagnoses were compared with those assigned by the consult nephrologists. RESULTS: Nephrologist A correctly diagnosed the cause of ARF in 24 of 26 samples (92.3% success rate) based on his performance of the UA. Diagnoses by nephrologists A and B, based on their review of the clinical laboratory UA report, were correct in only 23.1% and 19.2% of the samples, respectively. Accuracy of diagnosis for nephrologist B improved to 69.3% when she reviewed UA reports from nephrologist A. Nephrologist A's review of urine sediment was significantly more accurate than interpretations by nephrologist A or B of clinical laboratory reports (sign test, P < 0.001). Nephrologist A reported a greater number of renal tubular epithelial (RTE) cells (P < 0.0001), granular casts (P = 0.0017), hyaline casts (P = 0.0233), RTE casts (P = 0.0008), and dysmorphic red blood cells. The laboratory noted a greater number of squamous cells (P = 0.0034). CONCLUSION: A nephrologist is more likely to recognize the presence of RTE cells, granular casts, RTE casts, and dysmorphic red blood cells in urine. The laboratory may be reporting RTE cells incorrectly as squamous epithelial cells. Nephrologist-performed UA is superior to laboratory-performed UA in determining the correct diagnosis. PMID- 16253722 TI - Kidney volume and plasma hepatocyte growth factor-transforming growth factor beta1 ratio among children with biliary atresia before and after liver transplantation: the reversibility of nephromegaly. AB - BACKGROUND: We previously showed a positive correlation between nephromegaly and plasma hepatocyte growth factor (HGF)/transforming growth factor beta1 (TGF beta1) ratio in children with biliary atresia. The purpose of this study is to examine the possible reversibility of nephromegaly in patients with biliary atresia. METHODS: We evaluated kidney volume in 13 patients with biliary atresia before and after liver transplantation, 6 patients with hepatoblastoma, and 26 healthy children. Plasma HGF and TGF-beta1 levels were determined for all children. RESULTS: We noted significant nephromegaly in children with biliary atresia before liver transplantation compared with healthy children and children after liver transplantation (P < 0.001 and P = 0.006 for intercepts, P = 0.064 and P = 0.753 for slopes by analysis of covariance, respectively). The highest plasma HGF levels and HGF/TGF-beta1 ratios and the lowest TGF-beta1 concentrations were found in children with biliary atresia before liver transplantation (P < 0.001). No statistically significant nephromegaly was observed in children with biliary atresia after liver transplantation or those with hepatoblastoma despite the presence of a mildly increased plasma HGF level and HGF/TGF-beta1 ratio. Plasma HGF/TGF-beta1 ratio correlated positively with degree of nephromegaly in all patients (r = 0.717; P < 0.001). CONCLUSION: Our data suggest that liver transplantation reverses the nephromegaly present in children with biliary atresia and that plasma HGF/TGF-beta1 ratio may be associated with the development of nephromegaly in patients with biliary atresia. PMID- 16253723 TI - A comprehensive assessment of renal function in patients with Gaucher disease. AB - BACKGROUND: Gaucher disease (GD) is caused by deficiency of acid beta glucocerebrosidase and is the most common lysosomal storage disease. Patients may have massive hepatosplenomegaly, severe bone disease, and, occasionally, pulmonary or neurological involvement. Although other storage diseases, such as Fabry disease, frequently affect the kidneys, reports of renal abnormalities in patients with GD are limited to case reports. Our aim was to perform a comprehensive evaluation of renal function in patients with GD. METHODS: Evaluation was performed at routine clinic visits and included blood pressure recording and renal ultrasound. Serum chemistries, urinalysis, urine electrolytes, total protein, and tubular proteinuria were assessed, and estimated glomerular filtration rate (GFR) was calculated. RESULTS: One hundred sixty-one patients underwent evaluation, including 26 children. GFR was significantly greater in patients with GD than in age- and sex-matched healthy controls (P = 0.01 in men, P < 0.001 in women, P = 0.003 in children). Subgroups of patients with markers of more severe disease had a greater GFR than other patients. No patient had decreased renal function. Significant proteinuria was found only in patients with such comorbidities as diabetes mellitus or multiple myeloma. No evidence of renal tubular abnormalities was found, and kidney sonographic appearance and size were normal. CONCLUSION: Despite the multiorgan nature of the disease, a systematic evaluation did not find renal abnormalities in patients with GD. Glomerular hyperfiltration was observed in a proportion of patients, particularly those with markers of more severe disease. This phenomenon does not seem to be associated with a subsequent decline in renal function. PMID- 16253724 TI - Outcomes of acute coronary syndrome in a large Canadian cohort: impact of chronic renal insufficiency, cardiac interventions, and anemia. AB - BACKGROUND: Chronic renal insufficiency (CRI) has been identified as an important risk factor for cardiac events. Studies in the United States reported decreased survival and decreased use of surgical and medical interventions after myocardial infarction in patients with CRI. METHODS: We studied the impact of renal function on health outcomes in a Canadian cohort of consecutive patients admitted with acute coronary syndrome (ACS) between October 1997 and October 1999. The study design is an observational cohort of 5,549 adult patients who survived to discharge with a discharge diagnosis of ACS. Renal function is classified into 4 levels: (1) normal, glomerular filtration rate (GFR) greater than 80 mL/min/1.73 m2 (>1.33 mL/s); (2) mild CRI, GFR of 60 to 80 mL/min/1.73 m2 (1.00 to 1.33 mL/s); (3) moderate CRI, GFR of 30 to 59 mL/min/1.73 m2 (0.50 to 0.98 mL/s); and (4) severe CRI, GFR less than 30 mL/min/1.73 m2 (<0.50 mL/s). The primary outcome is death. RESULTS: Advanced and moderate CRI independently predicted death (hazard ratio, 1.06; 95% confidence interval [CI], 1.01 to 1.12; and hazard ratio, 1.23; 95% CI, 1.18 to 1.29). Severe anemia (hemoglobin level < 9.0 g/dL [<90 g/L]) also was an independent risk factor for death (hazard ratio, 1.38; 95% CI, 1.18 to 1.61). Use of beta-blockers (hazard ratio, 0.91; 95% CI, 0.86 to 0.97), acetylsalicylic acid (hazard ratio, 0.90; 95% CI, 0.84 to 0.97), lipid lowering therapy (hazard ratio, 0.84; 95% CI, 0.78 to 0.89), and medical thrombolysis (hazard ratio, 0.89; 95% CI, 0.81 to 0.97) were associated with reduced risk for death. Medical interventions with beta-blockers, acetylsalicylic acid, lipid-lowering therapy, and thrombolysis and surgical intervention were significantly less likely to be used in patients with CRI. CONCLUSION: Despite universal access to health care, Canadian patients with CRI are more likely to die after a cardiac event and less likely to receive important interventions. PMID- 16253725 TI - Association between carotid artery intima-media thickness and cardiovascular risk factors in CKD. AB - BACKGROUND: Common carotid artery intima-media thickness (CCA-IMT) correlates with cardiovascular events in the general population and is an independent predictor of cardiovascular mortality in the hemodialysis population. It has not been evaluated extensively in patients with chronic kidney disease. METHODS: CCA IMT was measured by using high-resolution B-mode ultrasonography, and glomerular filtration rate (GFR) was measured by means of EDTA clearance. Cardiovascular risk factors assessed included homocysteine and lipoprotein(a) levels, as well as smoking, blood pressure, and cholesterol level. RESULTS: One hundred fourteen patients were studied; mean measured GFR was 29.6 +/- 18.4 mL/min/1.73 m2 (0.49 +/- 0.31 mL/s). CCA-IMT was significantly elevated (0.59 +/- 0.22 cm) compared with a control group (0.44 +/- 0.08 cm; P = 0.0012). CCA-IMT increases with age (P < 0.0001) and low-density lipoprotein level (P = 0.048) and decreases with high-density lipoprotein level (P = 0.001) and being white (P = 0.014). CONCLUSION: This study suggests that arterial changes occur early in the course of renal disease progression and may be related to dyslipidemia in the early stages. PMID- 16253726 TI - The natural history of chronic renal failure: results from an unselected, population-based, inception cohort in Sweden. AB - BACKGROUND: Mortality rates in patients with chronic renal failure (CRF) are high both before and after start of renal replacement therapy (RRT). However, few studies of mortality and progression have been performed in an unselected CRF population. METHODS: We followed up a population-based inception cohort of 920 men and women aged 18 to 74 years who had CRF (serum creatinine level > 3.4 mg/dL [>300 micromol/L] for men and >2.8 mg/dL [>250 micromol/L] for women) for 55 to 79 months. Relationships between the outcomes (death and start of RRT) and independent variables under study (age, sex, primary renal disease, body mass index [BMI], and glomerular filtration rate [GFR] at entry) were explored by using Cox regression models. RESULTS: Seven hundred thirty-nine patients (80%) started RRT during the follow-up period. As expected, GFR at entry was clearly linked to the incidence of RRT (P < 0.0001). Age was related inversely to incidence of RRT (adjusted relative risk for patients > or =65 years relative to patients <45 years, 0.72; 95% confidence interval, 0.57 to 0.90). Men progressed to RRT more often than women (adjusted relative risk, 1.59; 95% confidence interval, 1.35 to 1.88). BMI was unrelated to RRT incidence. By the end of follow up, 389 patients with CRF (42%) had died, 89 of them (10%) before the start of RRT. The most common primary cause of death was cardiovascular disease (37.5%). Characteristics significantly related to a greater mortality rate included older age, diagnoses of diabetic nephropathy and nephrosclerosis, and low BMI. CONCLUSION: Preuremic characteristics (age, sex, primary renal diagnosis, BMI, and GFR) are predictive of prognosis in unselected patients with CRF. PMID- 16253727 TI - Association between body mass index and CKD in apparently healthy men. AB - BACKGROUND: Overweight and obesity are well-established risk factors for cardiovascular disease and decline in kidney function in individuals with existing chronic kidney disease (CKD). Conversely, their association with the development of CKD is less clear. METHODS: We evaluated the association between body mass index (BMI) and risk for CKD in a cohort of 11,104 initially healthy men who participated in the Physicians' Health Study and provided a blood sample after 14 years. BMI was calculated from self-reported weight and height. We estimated glomerular filtration rate (GFR) by using the abbreviated equation from the Modification of Diet in Renal Disease Study and defined CKD as GFR less than 60 mL/min/1.73 m2 (<1 mL/s/1.73 m2). RESULTS: After an average 14-year follow-up, 1,377 participants (12.4%) had a GFR less than 60 mL/min/1.73 m2 (<1 mL/s/1.73 m2). Higher baseline BMI was associated consistently with increased risk for CKD. Compared with participants in the lowest BMI quintile (<22.7 kg/m2), those in the highest quintile (>26.6 kg/m2) had an odds ratio (OR) of 1.45 (95% confidence interval [CI], 1.19 to 1.76; P trend <0.001) after adjusting for potential confounders. We found similar associations by using different categories of BMI. Compared with men who remained within a +/-5% range of their baseline BMI, those who reported a BMI increase greater than 10% had a significant increase in risk for CKD (OR, 1.27; 95% CI, 1.06 to 1.53). CONCLUSION: In this large cohort of initially healthy men, BMI was associated significantly with increased risk for CKD after 14 years. Strategies to decrease CKD risk might include prevention of overweight and obesity. PMID- 16253728 TI - Ultralate referral and presentation for renal replacement therapy: socioeconomic implications. AB - BACKGROUND: Clinical and metabolic complications of late referral (LR) for dialysis therapy have been well documented, but there is a paucity of data on its socioeconomic implications. This study examines the role of lifestyle and socioeconomic status on referral pattern. METHODS: During a 4-year period (1999 to 2002), we retrospectively reviewed records of all patients who initiated dialysis therapy at an urban tertiary-care center. Patients were classified into 3 categories according to the interval between first contact with a nephrologist and initiation of dialysis therapy: contact time of 3 months or longer indicates early referral (ER); 1 to less than 3 months, LR; and less than 1 month, ultralate referral (ULR). RESULTS: Of 460 patients (97% African Americans, 3% Hispanics), 212 patients (46%) were ULR, 168 patients (37%) were LR, and 80 patients (17%) were ER. Compared with ER and LR patients, those with ULR had significantly (P < 0.0001) lower hematocrits (23% versus 29% and 27%), serum albumin levels (3.1 versus 3.3 and 3.2 g/dL [31 versus 33 and 32 g/L]), and glomerular filtration rates (5 versus 8 and 7 mL/min/1.73 m2 [0.08 versus 0.13 and 0.12 mL/s/1.73 m2]), but greater rates of temporary dialysis catheter use (92% versus 39% and 70%) and mortality (40% versus 15% and 26%, respectively). Logistic regression analysis showed an association between mortality and homelessness (odds ratio, 3.8; P < 0.0001), polysubstance abuse (odds ratio, 2.3; P = 0.013), and alcoholism (odds ratio, 2.2; P = 0.009). Alcoholics (odds ratio, 2.5; P = 0.03), substance abusers (odds ratio, 5.5; P = 0.001), and the homeless/unemployed (odds ratio, 6.0; P = 0.004) were more likely to present as ULR cases. Patient-provided explanations for LR and ULR were denial (45%), unawareness of the presence of chronic kidney disease (30%), and economic difficulties (25%). Denial was more prevalent in LR (52%; P = 0.003) and ULR cases (39%; P = 0.003). CONCLUSION: Poor socioeconomic status is a major contributor to delayed referral. More efforts need to be directed at patient and physician chronic kidney disease educational awareness and improved health care access for inner-city and minority populations. PMID- 16253729 TI - Effect of comorbidity on the increased mortality associated with early initiation of dialysis. AB - BACKGROUND: Current recommendations for initiating dialysis therapy are based on level of kidney function and clinical evidence of uremia. Several studies reported no benefit in patient survival from initiating dialysis therapy with a greater glomerular filtration rate (GFR). Whether this is explained by a greater comorbidity burden or detrimental effect of early initiation remains unclear. We thus undertook an evaluation of the impact of comorbidity on the association between GFR at initiation and death. METHODS: Data from the Center for Medicare & Medicaid Services were used to derive 3 incident dialysis populations: (1) general population aged 18+ years, (2) older patients aged 67+ years, and (3) a "low-risk" subgroup without diabetes, heart failure, or atherosclerotic heart disease. A Cox proportional hazard regression technique was used. RESULTS: Greater GFR at initiation of dialysis therapy was associated with a greater risk for death in all populations, and sequential adjustment for additional covariates attenuated the effect. Patients in the general dialysis population who initiated dialysis therapy at a GFR greater than 10 mL/min/1.73 m2 (>0.17 mL/s) had a 42% increased risk for death compared with patients with a GFR less than 5 mL/min/1.73 m2 (<0.08 mL/s) at initiation of dialysis therapy after adjusting for all covariates. In the older and healthier populations, adjusted increased risks were 25% and 39%, respectively. CONCLUSION: Patients initiating dialysis therapy at greater GFRs have an increased risk for death not fully explained by comorbidity. Additional research is required to determine the reasons for poor survival in patients who start dialysis therapy with significant residual renal function. PMID- 16253730 TI - Prevalence of atrial fibrillation and associated factors in a population of long term hemodialysis patients. AB - BACKGROUND: Hemodialysis (HD) is associated with cardiovascular structural modifications; moreover, during HD, rapid electrolytic changes occur. Both factors may favor the onset of atrial fibrillation. METHODS: To define the prevalence of atrial fibrillation and identify associated factors, 488 patients on long-term HD therapy (age, 66.6 +/- 13.4 years; men, 58.0%; duration of HD, 76.5 +/- 84.3 months) were studied. RESULTS: Atrial fibrillation was reported in 27.0% of patients; paroxysmal in 3.5%, persistent in 9.6%, and permanent in 13.9%. Clinical and echocardiographic variables were considered: patients with atrial fibrillation were older (71.8 +/- 9.3 versus 64.7 +/- 14.2 years; P < 0.01), and its prevalence increased with age. Patients with arrhythmia had a longer duration of dialysis therapy (93.2 +/- 100.5 versus 70.2 +/- 76.7 months; P = 0.02). Atrial fibrillation was associated significantly with ischemic heart disease (P < 0.01), dilated cardiomyopathy (P < 0.01), acute pulmonary edema (P < 0.05), valvular disease (P < 0.05), cerebrovascular accidents (P < 0.05), and predialytic hyperkalemia (P < 0.05). Patients with atrial fibrillation more frequently showed left atrial dilatation (59.8% versus 34.5%; P < 0.0001), and in these subjects, left ventricular ejection fraction was significantly lower (53.9% versus 57.4%; P = 0.029). No association was found between arrhythmia and hypertension or diabetes. Multivariate analysis confirmed that patient age (P < 0.001), duration of HD therapy (P = 0.001), and left atrial dilatation (P < 0.001) were associated with atrial fibrillation. CONCLUSION: Atrial fibrillation is much more frequent in HD patients than in the general population; age, duration of HD history, presence of some heart diseases, and left atrial dilatation are associated with the arrhythmia. PMID- 16253731 TI - Citrate kinetics in patients receiving long-term hemodialysis therapy. AB - BACKGROUND: Regional anticoagulation using sodium citrate is used increasingly in hemodialysis patients at high risk for bleeding. However, citrate metabolism has never been evaluated systematically in hemodialysis patients, and it remains to be shown that citrate is cleared adequately in the presence of renal dysfunction. This study compares the pharmacokinetics of citrate in hemodialysis patients with minimal residual function with that in patients with normal renal function. METHODS: Long-term hemodialysis patients (n = 7) and patients without renal failure (n = 11) were investigated during routine immunoadsorption treatment by using a standardized citrate infusion protocol. Serial analysis of blood samples was performed before, during, and up to 120 minutes after citrate infusion (0.33 mmol/kg/h). Citrate plasma concentrations were measured colorimetrically. In addition, ionized calcium, pH, and bicarbonate were measured by using a blood gas analyzer. RESULTS: Basal (0.09 +/- 0.03 versus 0.12 +/- 0.03 mmol/L; P = not significant) and peak citrate concentrations were similar in both groups (1.24 +/ 0.42 versus 1.19 +/- 0.33 mmol/L; P = not significant). Citrate clearance was similar in patients with renal failure (0.31 +/- 0.06 L/min) and controls (0.35 +/- 0.11 L/min; P = 0.47). Effects on pH were minimal and did not differ between groups. No patient developed complications from citrate infusion. CONCLUSION: Compared with controls, citrate clearance and metabolism in long-term hemodialysis patients is not impaired, and no significant acid-base disorder occurred during citrate anticoagulation. From these data, it is tempting to conclude that citrate anticoagulation can be used safely in patients with chronic renal failure on regular hemodialysis therapy. PMID- 16253732 TI - Continuous venovenous hemofiltration with citrate-based replacement fluid: efficacy, safety, and impact on nutrition. AB - BACKGROUND: Citrate-based continuous venovenous hemofiltration (CVVH) replacement fluids provide effective and simple regional anticoagulation. However, concern over toxicity has limited citrate use, especially at the high filtration rates advocated for better outcomes. We used volumes of 72 L/d in patients at high risk for bleeding and investigated the treatment's efficacy, safety, and clinical results, especially with regard to nutrition supplementation. METHODS: A standard replacement solution (trisodium citrate, 13.3 mmol/L) was infused at up to 3 L/h in predilution CVVH, and ultrafiltration was increased further for net fluid removal. Calcium was repleted centrally. We retrospectively evaluated metabolic control, citrate toxicity, circuit patency, hemorrhagic complications, hemodynamics, vasopressor use, nutrition, renal recovery, and mortality. RESULTS: Seventy-six patients with 766 CVVH patient-days were analyzed. Mean replacement fluid rate was 31 mL/kg/h (35 mmol/h of citrate), with hemofiltration of 35 mL/kg/h (67 +/- 11 L/d). No significant bleeding, citrate toxicity, or hypocalcemia was observed, and 74% required additional alkali therapy. Dialyzer patency was 58% at 48 hours. Control of fluid, electrolytes, and azotemia was excellent (serum creatinine level, 1.7 mg/dL [150 micromol/L]; blood urea nitrogen, 42 mg/dL [15 mmol/L]). Fluid removal permitted protein (1.7 g/kg/d) and calorie (30 kcal/kg/d) nutrition in high fluid volumes. Vasopressor use and central pressures decreased significantly. Cumulative 28-day intensive care unit survival was 58%, and 41% of these patients had renal recovery in the intensive care unit. Thirty percent of the entire cohort survived the hospitalization, and 53% of these patients recovered renal function. CONCLUSION: CVVH with 3 L/h of citrate-based replacement fluid is a safe, efficient, and simple technique in patients at high risk for bleeding. It allows superb control of uremia and fluid balance and thereby permits aggressive nutritional support. PMID- 16253733 TI - Validation of 2 depression screening tools in dialysis patients. AB - BACKGROUND: Depression is the most common psychiatric disorder in long-term dialysis patients and is a risk factor for morbidity and mortality. An efficient and valid method of diagnosing depression might facilitate recognition and treatment. We sought to validate 2 depression assessment tools, the 21-question Beck Depression Inventory (BDI) and the 9-question Patient Health Questionnaire (PHQ-9), in a dialysis population. METHODS: We surveyed patients who had received dialysis for at least 90 days in Portland, OR. We excluded patients with dementia, delirium, or a history of major psychiatric disorders other than depression. The Structured Clinical Interview for Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, a gold-standard measure for depression, was administered by trained psychologists within 2 weeks of the BDI and PHQ-9. RESULTS: Of 62 enrolled subjects, 16 were diagnosed with a depressive disorder, including 12 patients (19%) with major depression, 3 patients with dysthymia, and 1 patient with minor depression. Optimal BDI and PHQ-9 cutoff values for depressive disorders combined was 16 or greater and 10 or greater, respectively. Sensitivities were 91% and 92%, specificities were 86% and 92%, positive predictive values were 59% and 71%, and negative predictive values were both 98%, with kappa values of 0.65 and 0.75, respectively. The difference between the 2 receiver operating characteristic curves was not statistically significant (P > 0.9). CONCLUSION: Our results validate the PHQ-9 and revalidate the BDI against a gold-standard measure for depressive disorders in the dialysis population. Both tools performed equally well. Because depression is prevalent, readily diagnosed, and associated with poor outcomes, screening by means of short and valid measurement tools may lead to better diagnosis and treatment of this modifiable risk factor. This may lead to improved clinical outcomes in dialysis patients. PMID- 16253734 TI - The Kidney Disease Outcomes Quality Initiative (K/DOQI) Guideline for Bone Metabolism and Disease in CKD: association with mortality in dialysis patients. AB - BACKGROUND: In 2003, the National Kidney Foundation-Kidney Disease Outcomes Quality Initiative (K/DOQI) published a guideline recommending tight control of serum calcium, phosphorus, calcium-phosphorus product (Ca x P), and intact parathyroid hormone levels in patients with chronic kidney disease. Within the context of this guideline, we explored associations of these plasma concentrations with all-cause mortality risk in incident dialysis patients in The Netherlands. METHODS: In a large, prospective, multicenter, cohort study (Netherlands Cooperative Study on the Adequacy of Dialysis), we included 1,629 patients new on hemodialysis or peritoneal dialysis therapy between 1997 and 2004. Multivariate Cox regression models containing calcium level, phosphorus level, intact parathyroid hormone level, age, comorbidity, primary kidney disease, nutritional status, albumin level, dialysis dose, and hemoglobin level were used to examine mortality risks. RESULTS: Mean age was 60 +/- 15 (SD) years, 61% were men, and 64% were treated with hemodialysis. In adjusted time-dependent survival analysis, all-cause mortality risk increased in hemodialysis patients by 40% (hazard ratio [HR], 1.4; 95% confidence interval [CI], 1.1 to 1.7) and in peritoneal dialysis patients by 60% (HR, 1.6; 95% CI, 1.1 to 2.4) for plasma phosphorus levels greater than the target. In addition, having elevated plasma Ca x P product levels increased mortality risk by 40% (HR, 1.4; 95% CI, 1.1 to 1.8) in hemodialysis patients and 50% in peritoneal dialysis patients (HR, 1.5; 95% CI, 1.0 to 2.2). In both patient groups, no significant effects were observed for plasma levels less than the targets. CONCLUSION: In time-dependent survival analysis, the presence of plasma phosphorus and Ca x P product concentrations greater than K/DOQI targets increased all-cause mortality risk in hemodialysis and peritoneal dialysis patients. PMID- 16253735 TI - Access flow in arteriovenous accesses by optodilutional and ultrasound dilution methods. AB - BACKGROUND: Most large studies evaluating the diagnostic properties of access blood flow (Qa) in arteriovenous (AV) accesses have used the Transonic HD01 (Transonic Systems Inc, Ithaca, NY) device, and recommended thresholds for angiography are based on data from these studies. There has been little exploration of how the use of other devices might affect the feasibility or performance of screening in AV accesses. METHODS: We compared 2 devices for measuring Qa: the Transonic HD01 and the Crit-Line TQA III (Hemametrics, Salt Lake City, UT). We studied 124 adults with end-stage renal disease and a functioning AV access (fistula or graft). Qa was measured with both devices in immediate succession during a single dialysis treatment. The primary outcome was the technical feasibility of the Qa measurement. We also compared mean Qa values measured by the Crit-Line III and Transonic devices. RESULTS: Qa measurements were less likely to be technically feasible when the Crit-Line III device was used compared with the Transonic device (86.3% versus 100%; P < 0.001). In patients with valid measurements, mean Qa measured using the Crit-Line III was significantly less than that measured using the Transonic HD01 device (886 +/- 557 versus 1,148 +/- 685 mL/min; P < 0.001). The mean difference was 261 mL/min (95% confidence interval [CI], 117 to 405) and was greater at higher levels of Qa. On average, Qa measured by means of the Crit-Line III device was 73% as high as that measured using the Transonic device (95% CI, 63 to 84). There was poor agreement between devices about whether criteria for angiography were met (kappa < 0.1). The proportion of patients for whom angiography was indicated (based on results from the Crit-Line device) was significantly greater than when only results from the Transonic device were considered (40.3% versus 7.3%; P < 0.001). CONCLUSION: Consideration should be given to device-specific Qa thresholds for angiography or, alternatively, standardization of Qa results between manufacturers. Clinicians should be aware that Qa results cannot be compared directly between different devices, and access monitoring should be performed using a single technique in any given patient. Additional studies are required before the Crit-Line TQA device can be recommended for widespread use. PMID- 16253736 TI - Mesenchymal conversion of mesothelial cells as a mechanism responsible for high solute transport rate in peritoneal dialysis: role of vascular endothelial growth factor. AB - BACKGROUND: During peritoneal dialysis (PD), the peritoneum is exposed to bioincompatible dialysis fluids that cause epithelial-to-mesenchymal transition of mesothelial cells, fibrosis, and angiogenesis. Ultrafiltration failure is associated with high transport rates and increased vascular surface, indicating the implication of vascular endothelial growth factor (VEGF). Sources of VEGF in vivo in PD patients remain unclear. We analyzed the correlation between epithelial-to-mesenchymal transition of mesothelial cells and both VEGF level and peritoneal functional decline. METHODS: Effluent mesothelial cells were isolated from 37 PD patients and analyzed for mesenchymal conversion. Mass transfer coefficient for creatinine (Cr-MTC) was used to evaluate peritoneal function. VEGF concentration was measured by using standard procedures. Peritoneal biopsy specimens from 12 PD patients and 6 controls were analyzed immunohistochemically for VEGF and cytokeratin expression. RESULTS: Nonepithelioid mesothelial cells from effluent produced a greater amount of VEGF ex vivo than epithelial-like mesothelial cells (P < 0.001). Patients whose drainage contained nonepithelioid mesothelial cells had greater serum VEGF levels than those with epithelial-like mesothelial cells in their effluent (P < 0.01). VEGF production ex vivo by effluent mesothelial cells correlated with serum VEGF level (r = 0.6; P < 0.01). In addition, Cr-MTC correlated with VEGF levels in culture (r = 0.8; P < 0.001) and serum (r = 0.35; P < 0.05). Cr-MTC also was associated with mesothelial cell phenotype. VEGF expression in stromal cells, retaining mesothelial markers, was observed in peritoneal biopsy specimens from high-transporter patients. CONCLUSION: These results suggest that mesothelial cells that have undergone epithelial-to-mesenchymal transition are the main source of VEGF in PD patients and therefore may be responsible for a high peritoneal transport rate. PMID- 16253737 TI - Allocation of deceased donor kidneys for transplantation: opinions of patients with CKD. AB - BACKGROUND: Deceased donor kidney allocation schemes are designed to balance optimal utility with equity of access. The aim of this single-center survey is to seek patient opinion about the relative importance of factors used to determine the optimal transplant recipient in kidney allocation schemes. METHODS: In each of 8 scenarios, participants were invited to select which 1 of 2 hypothetical patients should receive a kidney. RESULTS: Two hundred thirty-two of 295 invited patients (78.6%) completed the questionnaire: 104 of 153 invited hemodialysis patients (68.0%) and 128 of 142 invited patients with functioning transplants (90.1%). Only 6.0% of participants agreed with current UK Transplant (UKT) and United Network for Organ Sharing (UNOS) allocation to a patient not yet on dialysis therapy who had been on the transplant waiting list longer than a patient already on dialysis therapy. Only 24.6% of participants agreed with the UKT and UNOS schemes that the transplant survival advantage associated with HLA matching warranted allocation of a kidney to a patient who had been waiting 2 years in preference to a patient waiting 7 years. Participants also were opposed to the use of recipient age and balance of exchange agreements (that reward centers with high rates of organ retrieval). The majority agreed with UKT and UNOS that recipient sex should not be used to allocate kidneys and allocation should favor recipients who have waited longer. CONCLUSION: Patients disagreed with several aspects of current allocation systems. Analysis of patient opinion should be taken into consideration when attempting to optimize the use of this scarce health resource. PMID- 16253738 TI - Malignant hypertension with intestinal ischemia secondary to juxtaglomerular cell tumor. AB - Malignant hypertension is a well-defined condition associated with high blood pressure and acute target-organ damage. Although 95% of cases are secondary to essential hypertension, its etiological profile is broad. Juxtaglomerular cell tumor is a rare condition, with only approximately 65 cases reported to date. We describe a patient with malignant hypertension with acute renal failure and intestinal ischemia secondary to a juxtaglomerular cell tumor. We believe this is the first case of juxtaglomerular cell tumor causing malignant hypertension. The diagnostic approach and treatment are discussed. PMID- 16253739 TI - Coronary artery bypass graft mycotic aneurysms in a dialysis patient. AB - Infection is a common problem in dialysis patients and ranks second behind cardiovascular disease as a major cause of death. The major causes of infections, mainly bloodstream infections, often are related to dialysis access. Metastatic infectious complications have been reported frequently in the course of such bacteremias. We report the case of a 79-year-old dialysis patient who was admitted with recurrent catheter-related bacteremia caused by methicillin resistant Staphylococcus aureus. Echocardiography and a computed tomographic scan of her chest showed multiple coronary artery bypass graft mycotic aneurysms. Despite prompt dialysis catheter removal and antibiotic treatment, she had progressive deterioration of her hemodynamic and mental status and eventually died of profound sepsis. An autopsy confirmed computed tomographic findings, plus extensive suppuration involving the left atrial and ventricular myocardium and upper lobe of the left lung. To our knowledge, this is the first report of coronary artery graft aneurysms complicating infective endocarditis in a dialysis patient. PMID- 16253740 TI - EPO therapy during acute kidney disease: to use or not to use, that is the question. PMID- 16253741 TI - Hypothesis versus association: the optimal hemoglobin target debate. PMID- 16253742 TI - The development of kidney allocation policy. PMID- 16253743 TI - Hemodialysis: techniques and prescription. PMID- 16253744 TI - Chronic renal failure and abnormal tubular cells in 2 siblings. PMID- 16253745 TI - Diagnostic discordance for hepatitis C virus infection in hemodialysis: correlations with clinical and laboratory features. PMID- 16253747 TI - Are homocysteine and MTHFR genotype polymorphism associated with arteriovenous fistula patency? PMID- 16253749 TI - Development of an oscillating/rotating/pulsating toothbrush: The Oral-B professionalCaretrade mark series. AB - The Oral-B range of power toothbrushes are based on a clinically proven oscillating/rotating action. This review considers the extensive series of laboratory and clinical studies involved in the development of the Oral-B power toothbrushes with three-dimensional (3D) brush head action (i.e. oscillating/rotating/pulsating), now known as the Oral-B ProfessionalCaretrade mark Series. The unique, highly effective cleaning performance of the 3D action incorporated in the D15 and D17 models is proven in both the laboratory and clinical environments, thereby establishing an evidence-based platform for the development of the most recent ProfessionalCare 7000. In the early development phase, in-vitro studies indicated that the additional pulsating action and advancements in filament technology improved approximal penetration, where plaque removal is most difficult. In-vitro models provide a useful development tool for the rapid assessment of design modifications, while controlling many factors that confound in-vivo studies, but the relevance of their findings needs to be confirmed in the clinical situation by direct improvements in oral health. Results from well-controlled, randomised clinical studies in the development programme have demonstrated the superiority of the D15 and D17 models with 3D action over a manual toothbrush and other power toothbrushes of various actions and designs in the improvement of oral health outcomes such as plaque removal (especially from approximal sites), reduction of gingival inflammation, and control of calculus and stain formation, with no greater potential to cause oral tissue abrasion. Furthermore, user acceptance of the oscillating/rotating/pulsating toothbrush may encourage long-term compliance during normal use. PMID- 16253750 TI - Single-use plaque removal efficacy of three power toothbrushes. AB - OBJECTIVES.: To compare the safety and plaque removal efficacy of two oscillating/rotating/pulsating toothbrushes (Oral-B ProfessionalCaretrade mark 7000 [PC 7000] and Oral-B 3D Excel [3DE]) and a high-frequency toothbrush (Sonicare(R) Advance, Philips Oral Healthcare; SA) in a single-use, examiner blind, three period crossover study. METHODS.: After refraining from all oral hygiene procedures for 23-25 hours, subjects received an oral tissue examination and those with pre-brushing whole mouth mean plaque scores 0.6 based on the Rustogi et al. Modified Navy Plaque Index were randomly assigned to treatment sequence. After brushing with the assigned toothbrush and a commercially available dentifrice for 2 minutes, oral tissues were then re-examined and post brushing plaque scores recorded. Following a brief washout period between two additional visits, the above procedures were repeated with the two alternate toothbrushes. One examiner, blinded to the treatment sequence, performed all clinical measurements. RESULTS.: A total of 79 subjects (28 males and 51 females) were enrolled and completed the study. Each toothbrush was found to be safe and significantly reduced plaque levels after a single brushing. The PC 7000 and 3DE were equally more effective in plaque removal than the SA, at all tooth areas, reducing plaque by 59.0%, 59.7% and 51.8%, respectively on whole mouth surfaces, and by 67.5%, 67.8% and 59.4%, respectively on approximal surfaces. CONCLUSIONS.: The action of the oscillating/rotating/pulsating toothbrushes (Oral-B ProfessionalCare 7000 and Oral-B 3D Excel) was more effective in plaque removal than the high-frequency toothbrush (Sonicare Advance). PMID- 16253751 TI - Plaque removal efficacy of two electric toothbrushes with different brush head designs. AB - OBJECTIVES.: To compare the safety and plaque removal efficacy of two electric toothbrushes, one a rechargeable oscillating/pulsating toothbrush with a small round brush head (Oral-B ProfessionalCaretrade mark 7000; PC 7000), the other a battery-operated toothbrush with a dual moving brush head (Crest(R) SpinBrushtrade mark Pro; SBP). METHODS.: The study had a randomised, examiner blind, two-arm crossover design. All subjects received an oral prophylaxis and used both toothbrushes on alternating days for a two-week practice period. After abstaining from all oral hygiene procedures for 23-25 hours, subjects received an oral tissue examination and those with pre-brushing whole mouth mean plaque scores >/=0.60 measured by the Rustogi et al. Modified Navy Plaque Index were randomly assigned to treatment sequence. Subjects brushed with their assigned toothbrush for 2 minutes using a commercially available dentifrice. Oral tissues were then re-examined and post-brushing plaque scores recorded. After a brief washout period, the above procedures were repeated with the alternate toothbrush. One examiner, blinded to the treatment sequence, performed all clinical measurements. RESULTS.: A total of 70 subjects (24 males and 46 females) were enrolled and completed the study. Each toothbrush was found to be safe and significantly reduced plaque levels after a single brushing. The PC 7000 was significantly more effective in plaque removal than the SBP at all tooth areas, reducing whole mouth plaque by 61% versus 58% and plaque from approximal surfaces by 69% versus 65%, respectively. CONCLUSIONS.: The action of the oscillating/pulsating toothbrush with a small round brush head, Oral-B ProfessionalCare 7000, is more effective in plaque removal than the battery operated Crest SpinBrush Pro toothbrush with a larger dual moving brush head. PMID- 16253752 TI - Three-month assessment of safety and efficacy of two electric toothbrushes. AB - OBJECTIVE.: This randomised, examiner-blind parallel group study was designed to evaluate the safety and efficacy of a rechargeable oscillating/pulsating toothbrush (Oral-B ProfessionalCaretrade mark 7000, Oral-B Laboratories; PC 7000) and a battery-operated toothbrush (Crest(R) SpinBrushtrade mark Pro, Procter & Gamble Company; SBP) in the reduction of gingivitis, bleeding and plaque over a 3 month period. METHODS.: After 12-18 hours of no oral hygiene, subjects had oral tissue examinations, and gingival and plaque assessments to determine eligibility in the study. Subjects were stratified and randomised into treatment groups based on initial whole mouth mean plaque (Turesky modification of Quigley Hein Plaque Index) and gingivitis (Loe & Silness Gingival Index) scores and gender. Subjects were instructed to brush twice daily with their assigned toothpaste and toothbrush. Clinical parameters were assessed at baseline, and after 1 and 3 months of use. Within treatment comparisons from baseline were analysed using t test; between treatment comparisons were analysed using ANOVA. RESULTS.: Data were obtained from 92 subjects (PC 7000 n=45; SBP n=47). No significant differences were found in baseline plaque, gingivitis and bleeding between groups. Both treatment groups had significant reductions from baseline in plaque, gingivitis and bleeding scores. PC 7000 demonstrated significantly greater reductions compared to SBP in whole mouth plaque at 1 month: 0.39+/-0.43 vs. 0.16+/-0.42 and 3 months: 0.32+/-0.48 vs. 0.04+/-0.41. PC 7000 also demonstrated significant reductions compared to SBP in gingivitis at 3 months for whole mouth: 0.14+/-0.09 vs. 0.10+/-0.10 and approximal areas: 0.11+/-0.08 vs. 0.08+/-0.09. There were no significant differences between toothbrushes in bleeding at either time point. Safety examinations revealed no apparent difference in soft and hard tissue abnormalities between groups. CONCLUSION.: The PC 7000 toothbrush demonstrated significantly greater reductions in plaque and gingivitis compared to the SPB over a 3-month period. PMID- 16253754 TI - Efficacy of a novel brush head in the comparison of two power toothbrushes on removal of plaque and naturally occurring extrinsic stain. AB - OBJECTIVES.: To compare the safety and efficacy of an oscillating/pulsating power toothbrush (Oral-B ProfessionalCaretrade mark 7000; PC 7000) fitted with either the standard FlexiSoft (PC 7000/EB17) brush head or the novel Pro Polisher (PC 7000/EB-Prophy) and a high-frequency toothbrush (Philips Sonicare(R) Elite(R); SE), in their relative ability to remove plaque and naturally occurring extrinsic dental stain over a six-week period. METHODS.: This randomised, examiner-blind, parallel group study involved 90 healthy subjects from a general population. All subjects received a baseline plaque (Turesky et al. modified Quigley-Hein Plaque Index), stain (Lobene Stain Index) and tooth shade (VITAPAN(R) Shade Guide) [Vita] assessment and an oral tissue examination. After training in the use of their randomly assigned device, subjects were instructed to brush twice daily for 2 min and returned after 3, 4 and 6 weeks of product use for a repeat of each clinical assessment. RESULTS.: Reductions from baseline in mean plaque and extrinsic dental stain scores were significant at Weeks 3, 4 and 6 in all three treatment groups. By Week 6, mean reductions from baseline in whole mouth plaque scores were 32%, 27% and 14% in the PC 7000/EB-Prophy, PC 7000/EB17 and SE groups, respectively. For the body of the tooth, mean reductions from baseline at Week 6 in total stain were 89%, 89% and 80%, respectively. Between treatment group comparisons consistently revealed that the PC 7000 toothbrush plus the EB Prophy or EB17 brush head removed significantly more plaque and extrinsic stain (total stain, stain area and stain intensity) than the SE toothbrush at 3, 4 and 6 weeks. The EB-Prophy group had a greater proportion of subjects showing a 2-3+ change in Vita shade scores at each time point compared to the other two brushes; at Week 6 the proportions were 67% in the PC 7000/EB-Prophy group, 30% in the PC 7000/EB17 group, and 7% in the SE group. The PC 7000/EB17, PC 7000/EB-Prophy and the SE were found to be safe as used in the study. CONCLUSIONS.: The oscillating/rotating/pulsating PC 7000 (fitted with either the standard EB17 or novel EB-Prophy brush head) is more effective at removal of plaque and naturally occurring extrinsic tooth stain, and the PC 7000 plus EB-Prophy in the improvement of tooth shade, than the high-frequency SE toothbrush. PMID- 16253755 TI - Effects of active MEK1 expression in vivo. AB - Cell transformation is often a result of constitutive activation of genes in signaling pathways that regulate cell proliferation and differentiation. Indeed, the Ras/Raf/MEK/ERK mitogen-activated protein kinase (MAPK) signaling pathway is constitutively activated in a large number of cancers. The extent to which a single-gene mutation can alter cell fate, however, remains questionable. In vitro studies have addressed this issue, but organs are comprised of multiple cell types, and in vitro models often poorly approximate these interactions. In response to these limitations, cell-type specific mouse models have been generated as a means to examine the effect of altering a single element of the MAPK pathway in vivo. This review summarizes data from transgenic murine and human tissue models expressing constitutive active forms of MEK1. PMID- 16253756 TI - Centrosome amplification, chromosome instability and cancer development. AB - During mitosis, two centrosomes form spindle poles and direct the formation of bipolar mitotic spindles, which is an essential event for accurate chromosome segregation into daughter cells. The presence of more than two centrosomes (centrosome amplification), severely disturbs mitotic process and cytokinesis via formation of more than two spindle poles, resulting in an increased frequency of chromosome segregation errors (chromosome instability). Destabilization of chromosomes by centrosome amplification aids acquisition of further malignant phenotypes, hence promoting tumor progression. Centrosome amplification occurs frequently in almost all types of cancer, and is considered as the major contributing factor for chromosome instability in cancer cells. Upon cytokinesis, each daughter cell receives one centrosome, and thus centrosome must duplicate once, and only once, before the next mitosis. If centrosomes duplicate more than once within a single cell cycle, centrosome amplification occurs, which is frequently seen in cells harboring mutations in some tumor suppressor proteins such as p53 and BRCA1. The recent studies have provided critical information for understanding how loss of these proteins allows multiple rounds of centrosome duplication. In this review, how centrosome amplification destabilizes chromosomes, how loss of certain tumor suppressor proteins leads to centrosome amplification, and the role of centrosome amplification in cancer development will be discussed. PMID- 16253757 TI - Association of human herpesvirus type 6 DNA with human bladder cancer. AB - We examined the presence of human herpesvirus type 6 (HHV6) DNA in a series of 74 bladder carcinomas from a Mediterranean population to elucidate their possible role as cofactor in the development of bladder cancer with or without associated human papillomavirus (HPV) infection. HHV-6 type B DNA was present in 5 men (6.8%) out of the 74 tumors investigated; two of them had associated HPV-16 DNA in the same specimen. In one case that had associated urothelial carcinoma in situ, both HHV-6B and HPV-16 DNA were present. In conclusion, the low incidence of HHV-6B in bladder cancer and the ubiquitous nature of HHV-6 infection are more consistent with a bystander role rather than cofactor in the oncogenesis of bladder cancer. PMID- 16253758 TI - Enhancement by acrylamide of N-methyl-N-nitrosourea-induced rat mammary tumor development-possible application for a model to detect co-modifiers of carcinogenesis. AB - Acrylamide (AA) has recently been reported to be spontaneously formed in fried and baked foods with various concentrations. Although carcinogenicity in humans is as yet equivocal, numerous positive genotoxicity data in vitro and in vivo and results of rat long-term carcinogenicity studies demonstrating tumor induction at multiple sites, like the mammary gland, thyroid and testes, suggest the risk with dietary exposure may not be negligible. In the present study, to establish a medium-term carcinogenesis model for screening of agents with the potential to modify AA effects on the mammary gland and thyroid, we pretreated rats with 7,12 dimethylbenz(a)anthracene (DMBA), in combination with N-bis(2 hydroxypropyl)nitrosamine (DHPN), or N-methyl-N-nitrosourea (MNU) alone and then administered AA at 20 and 40 ppm in the drinking water for 30 weeks. The incidence and multiplicity of mammary tumors were increased at the high dose (P<0.05) in MNU- but not DMBA+DHPN-treated rats. No thyroid tumors were induced in any case. The results indicate that the MNU model is suitable for detection of modifiers of AA actions. PMID- 16253759 TI - Cooperative cell-growth inhibition by combination treatment with ZD1839 (Iressa) and trastuzumab (Herceptin) in non-small-cell lung cancer. AB - An important recent advance in anticancer therapy was the development of molecular-targeting drugs, such as the epidermal growth-factor receptor (EGFR) targeting drug ZD1839 (Iressa) and the HER2-trageting anti-HER2 monoclonal antibody trastuzumab (Herceptin). ZD1839 and trastuzumab are reported to improve the therapeutic efficacy of treatment for non-small-cell lung cancer (NSCLC) and breast cancer, respectively, although the effectiveness of either drug alone is not satisfactory. NSCLC cells often express both EGFR and HER2. We therefore investigated whether a combination of ZD1839 and trastuzumab had an additive or synergistic antitumor effect. In culture ZD1839 inhibited the growth of four NSCLC cell lines (A549, NCI-H23, NCI-H727, and NCI-H661) that expressed various levels of EGFR, HER2, HER3, and HER4. A significant cytotoxic effect was observed when ZD1839 was combined with trastuzumab in A549 cells. However, this combination had no apparent effect in NCI-H23 cells. Significant G(1)-phase arrest, increased p27 expression and decreased cyclin E or D1 levels were detected in A549 cells treated with ZD1839 and trastuzumab. No significant effects were detected in NCI-H23 cells examined. The combination treatment significantly inhibited the phosphorylation of EGFR, HER2, retinoblastoma, extracellular signal-regulated kinase-1/2, and protein kinase B/Akt in A549 cells, but not in NCI-H23 cells. Our results indicated that increased levels of constitutive EGFR/HER2 heterodimers were formed in A549 cells in the presence of ZD1839, whereas no heterodimer formation was detected in NCI-H23 cells. We therefore suggest that combination treatment with ZD1839 and trastuzumab might have improved therapeutic efficacy against NSCLC cells expressing both EGFR and HER2. PMID- 16253761 TI - Specific induction of oxidative stress in terminal bronchiolar Clara cells during dimethylarsenic-induced lung tumor promoting process in mice. AB - The induction of oxidative stress in pulmonary cells during the process of lung tumor promotion by dimethylarsinic acid (DMA), a main metabolite of inorganic arsenics in mammals, was examined by immunohistochemical analysis using a specific antibody against 4-hydroxy-2-nonenal (4HNE) adducts, which are major aldehydic metabolites of lipid peroxidation. We demonstrated that 4HNE-modified proteins existed specifically in the secretory granules in terminal bronchiolar Clara cells. Furthermore, the degree of positive staining increased with the duration of DMA administration. Transmission electron microscopy revealed morphological changes in the Clara cells of DMA-treated mice. These results suggest that Clara cells are the major target cell for DMA-induced oxidative stress and that the cells may play an important role in the lung tumor promotion process in mice. PMID- 16253760 TI - Stimulation of cultured colon 26 cells with TNF-alpha promotes lung metastasis through the extracellular signal-regulated kinase pathway. AB - We investigated the influence of TNF-alpha on the metastasis of cancer cells. Treatment of cultured colon 26 cells with TNF-alpha enhanced metastatic properties including production of MMP-9, adhesion, migration and invasion. Cells treated with TNF-alpha in vitro showed marked potential to metastasize to the lung and liver in vivo. U0126, an inhibitor of MEK1/2, inhibited the TNF-alpha induced activation of extracellular signal-regulated kinase 1/2 (ERK1/2) and the metastatic properties in vitro without affecting cell proliferation. In addition, pretreatment with U0126 in vitro completely abrogated the increased lung metastasis of TNF-alpha-treated cells. These results indicate that TNF-alpha induced activation of cancer cells through the ERK pathway is sufficient for the enhanced metastatic potential of colon 26 cells. PMID- 16253762 TI - A novel loss-of-function deletion in sodium/iodide symporter gene in follicular thyroid adenoma. AB - The sodium/iodide symporter (NIS) actively transports iodide into thyrocytes. However, in thyroid carcinoma, down-regulated or mis-targeted NIS expression is commonly found and usually correlates with tumor dedifferentiation and loss of radioiodine uptake capacity. In this study, we screened NIS genes of thyroid tumor tissues from three patients with thyroid carcinoma by using reverse transcription-polymerase chain reaction and nucleotide sequencing. We found a novel exon 6 deletion in NIS gene. We then examined the NIS gene from the blood of this patient. The nucleotide sequences of the flanking region of exon 6 were normal. By transient transfection and I-125 uptake assay, we found that the wild type NIS-expressing HepG2 cells accumulated six times more iodide than mutant and mock HepG2 cells. Our data demonstrated that the exon 6 deletion causes an iodide trapping defect. PMID- 16253763 TI - Regulation of human peptide transporter 1 (PEPT1) in gastric cancer cells by anticancer drugs. AB - Human peptide transporter 1 (PEPT1) mediates the cellular uptake of di- and tripeptides and peptide-like drugs in the small intestine. In the present study, we examined the regulation of PEPT1 by anticancer drugs in the gastric cancer cell line MKN45. PEPT1 was expressed and functioned in MKN45 cells. The transport activity and mRNA expression of the facilitative glucose transporter 1 (GLUT1) were significantly decreased by 5-fluorouracil treatment, but those of PEPT1 were slightly increased. Cisplatin treatment affected neither PEPT1 nor GLUT1 activity. In conclusion, PEPT1 expressed in MKN45 cells are resistant against the cellular injury induced by 5-fluorouracil and cisplatin. PMID- 16253764 TI - Characterization of two new variants of human catechol O-methyltransferase in vitro. AB - Catechol O-methyltransferase (COMT) plays an important role in the inactivation of biologically active and toxic catechols. It has been shown that human soluble COMT (S-COMT) is genetically polymorphic with a wild type and at least one variant in which a valine has been substituted with a methionine at codon 108. This polymorphism has been the subject of intense molecular epidemiological studies because of the important role of COMT in the metabolism of catecholamines and catechol estrogens. Several epidemiological studies have shown that women, homozygous with the Val108Met variant, have an increased risk of developing estrogen-associated cancers. However, some other studies have shown that this COMT polymorphism is not associated with increased risk of developing cancers. These conflicting data suggest that additional COMT genetic variants might contribute to the increased risk of developing cancers. Although two new single nucleotide polymorphisms (SNP) that cause amino acid substitutions Ala22Ser and Ala52Thr have been identified recently, they have not been fully characterized. In the present study, Ala22Ser and Ala52Thr variants of human S-COMT were produced using recombinant DNA techniques, and then COMT properties were measured including enzymatic activity, thermostability, and sensitivity to inhibition mediated by 4-hydroxyequilenin (4-OHEN). The Ala22Ser variant showed lower methylation capacity and higher thermolability. In addition, this variant is sensitive to 4-OHEN mediated irreversible inhibition. Our data indicate that the Ala22Ser polymorphism might also be of functional significance and might play a role in susceptibility to estrogen-associated cancers. PMID- 16253765 TI - Diphenylmethyl selenocyanate inhibits DMBA-croton oil induced two-stage mouse skin carcinogenesis by inducing apoptosis and inhibiting cutaneous cell proliferation. AB - Numerous epidemiological and experimental studies have showed the inverse relationship between dietary selenium intake and different types of cancer. Continuous efforts are going on to develop suitable organoselenium compounds, which can be used as cancer chemopreventive agents for human. In the present study, a synthetic organoselenium compound diphenylmethyl selenocyanate was evaluated for its ability to arrest cell proliferation and to induce apoptosis against 7,12-dimethylbenz[a]anthracene-croton oil induced two-stage mouse skin carcinogenesis model. Reduction in the incidence and number of papilloma, the preneoplastic lesion, was considered to be the mean of assessment. Significant decrease in the level of cell proliferation (p<0.01) and significant enhancement in the level of apoptosis (p<0.01) were found. Caspase-3, which contribute a part in the process of cellular apoptosis to prevent further cellular differentiation was also elevated significantly (P<0.01) during the treatment with the Se compound. These observations seem to be correlated with the significant reduction in the corresponding number of skin papilloma formation after 12 weeks of experiment. Thus the compound, diphenylmethyl selenocyanate may be considered for further research to establish it as an effective cancer chemopreventive agent. PMID- 16253766 TI - Apoptotic activity of isomalabaricane triterpenes on human promyelocytic leukemia HL60 cells. AB - Four isomalabaricane triterpenes were isolated from marine sponge Geodia japonica [W.H. Zhang, C.T. Che, Isomalabaricane-type nortriterpenoids and other constituents of the marine sponge Geodia japonica, J. Nat. Prod. 64 (2001) 1489 1492. ] and their cytotoxicity was evaluated using a human promyelocytic leukemia HL60 cell line. Of the four triterpenes tested, geoditin A was the most cytotoxic to HL60 cells [IC50=3 microg/ml (<6.6 microM)], followed by stellettins A and B, whereas geoditin B exhibited relatively weak cytotoxicity. The treated cells manifested nuclear changes characteristic for apoptosis, and associated with dissipation of mitochondrial membrane potential, activation of caspase 3, and decrease of cytoplasmic proliferating cell nuclear antigen (PCNA), as demonstrated by fluorescence and immunofluorescence microscopy. When the HL60 cells were exposed to geoditin A ranging from 1.25 to 25 microg/ml, a dose dependent increase of reactive oxygen species, a progressive dissipation of mitochondrial membrane potential, and an increase in annexin V-FITC binding were measured by flow cytometry. Taken together our results suggest that geoditin A markedly induced reactive oxygen species, decreased mitochondrial membrane potential and mediated a caspases 3 apoptosis pathway. PMID- 16253767 TI - Induction of apoptosis in human leukemia cells by black tea and its polyphenol theaflavin. AB - Treatment of human leukemic cell lines HL-60 and K-562 with extracts of green and black tea and their polyphenols epigallocatechin gallate and theaflavins, respectively, showed a dose dependent inhibition of growth as a result of cytotoxicity and suppression of cell proliferation. Based on the IC50 values obtained from cytotoxicity data it was clearly evident that black tea was as efficient as green tea. Analysis of polyphenol contents of tea extracts revealed that not only epigallocatechin gallate, which is a predominant polyphenol of green tea, but also theaflavin that is abundantly present in black tea affords significant chemotherapeutic action by imparting cytotoxicity to human leukemic cells. Electrophoretic analysis of fragmented DNA from treated cells displayed characteristic ladder pattern. Flow cytometric analysis revealed the dose dependent increase in sub-G1 peak. These criteria confirmed that cytotoxic activity of green and black tea was due to induction of apoptosis. Such induction was found to be mediated through activation of caspases 3 and 8, particularly caspase 3 and by altering apoptosis related genes as evident by down-regulation of Bcl-2 and up-regulation of Bax proteins. PMID- 16253768 TI - Glucose requirement for hypoxic accumulation of hypoxia-inducible factor-1alpha (HIF-1alpha). AB - Hypoxia-inducible factor-1alpha (HIF-1alpha) is both a potential endogenous marker of tumor hypoxia and therapeutic target, and elevated HIF-1alpha protein levels have been shown to be associated with increased hypoxic radiation resistance in FaDu human pharyngeal carcinoma cells in vitro. Here, we found that in FaDu xenografts, no significant HIF-1alpha protein accumulation was detectable by either flow cytometry or Western blot, despite the presence of hypoxic (pimonidazole-positive, radiation resistant) cells. To investigate the effect of different tumor microenvironment conditions on hypoxic HIF-1alpha accumulation, we performed in vitro hypoxia experiments (0.1% O2, 24 h) with manipulation of pH (7.4 vs. 6.7), glucose (0-5.5 mM) and serum (0 or 10%) availability in FaDu and HT 1080 human fibrosarcoma cells. Hypoxic induction of HIF-1alpha protein was strongly dependent on glucose availability and largely abolished at 0.55 mM glucose or less in both cell lines. This glucose effect was confirmed in a hypoxia-responsive-element (HRE)/enhanced-green-fluorescent-protein (EGFP) reporter assay in transfected HT 1080 cells and possibly explains a lack of HIF 1alpha protein in hypoxic tumor cells. PMID- 16253769 TI - Water-soluble polymeric polyphenols from cinnamon inhibit proliferation and alter cell cycle distribution patterns of hematologic tumor cell lines. AB - To explore possible anti-cancer properties of water-soluble, polymeric polyphenols from cinnamon, three myeloid cell lines (Jurkat, Wurzburg, and U937) were exposed to increasing concentrations of an aqueous extract prepared from cinnamon (CE) for 24 h. Cell growth and cell cycle distribution patterns responded in a dose-dependent manner to CE. That is, an increase in the percentage of cells distributed in G2/M was observed in all three cell lines as the amount of CE increased. At the highest dose of CE, the percentage of Wurzburg cells in G2/M was 1.5- and 2.0-fold higher than those observed for Jurkat and U937 cells, respectively. Wurzburg cells lack the CD45 phosphatase and may be more sensitive to imbalances in signaling through kinase/phosphatase networks that promote growth. The results suggest the potential of CE to interact with phosphorylation/dephosphorylation signaling activities to reduce cellular proliferation in tandem with a block at the G2/M phase of the cell cycle. PMID- 16253770 TI - beta-Catenin, Nf-kappaB and FAS protein expression are independent events in head and neck cancer: study of their association with clinical parameters. AB - In spite of much effort, no good markers have yet been found for predicting prognosis or response to therapy in advanced head and neck squamous cell carcinoma (HNSCCs) patients. beta-catenin, a protein involved in the cytoskeleton, cell-cell adhesion and gene transcription, is a factor associated with tumour progression. Recently, an interaction has been reported between beta catenin, and NF-kappaB coupled with an inverse association of beta-catenin, and FAS (CD95/APO-1) protein expression in breast and colorectal tumours. To confirm these observations and to test their clinical impact in HNSCCs we have evaluated the expression of beta-catenin, NF-kappaB and FAS proteins. We used tissue microarrays to simultaneously analyse the levels of these proteins immunohistochemically in 118 HNSCCs. Among the 113 tumours evaluable for beta catenin, increased and decreased levels were detected in 41 (36%) and 62 (55%) of the tumours, respectively. beta-catenin, protein staining was mainly membranous but 10 tumours (9%) showed the clear presence of protein in the cytoplasm, and none in the nucleus. Moreover, 81% of the tumours had decreased FAS protein expression, indicating that loss of FAS protein is a common feature of HNSCCs. Abnormal or nuclear NF-kappaB staining was observed in 24% of the tumours. No association was detected between the expression levels of the proteins evaluated. Regarding clinical associations, tumours from the hypopharynx had significantly lower levels of beta-catenin expression than those from other locations (P<0.05). Moreover, our data revealed that patients whose tumours had low levels of beta catenin protein expression had decreased survival probability (24.8 months vs. NR, P=0.03) and reduced response to therapy (15.4 vs. 43 months; P=0.01) compared with patients whose tumours had high levels of beta-catenin. Taken together, our observations indicate that beta-catenin, NF-kappaB and FAS expression are independent events during HNSCC development and that levels of beta-catenin protein may identify subsets of advanced HNSCCs patients with different prognosis and response to therapy capabilities. PMID- 16253771 TI - Reply to: Low prevalence of BRAF mutations in radiation-induced thyroid tumors in contrast to sporadic papillary carcinomas. PMID- 16253772 TI - Automatic prostate localization on cone-beam CT scans for high precision image guided radiotherapy. AB - PURPOSE: Previously, we developed an automatic three-dimensional gray-value registration (GR) method for fast prostate localization that could be used during online or offline image-guided radiotherapy. The method was tested on conventional computed tomography (CT) scans. In this study, the performance of the algorithm to localize the prostate on cone-beam CT (CBCT) scans acquired on the treatment machine was evaluated. METHODS AND MATERIALS: Five to 17 CBCT scans of 32 prostate cancer patients (332 scans in total) were used. For 18 patients (190 CBCT scans), the CBCT scans were acquired with a collimated field of view (FOV) (craniocaudal). This procedure improved the image quality considerably. The prostate (i.e., prostate plus seminal vesicles) in each CBCT scan was registered to the prostate in the planning CT scan by automatic 3D gray-value registration (normal GR) starting from a registration on the bony anatomy. When these failed, registrations were repeated with a fixed rotation point locked at the prostate apex (fixed apex GR). Registrations were visually assessed in 3D by one observer with the help of an expansion (by 3.6 mm) of the delineated prostate contours of the planning CT scan. The percentage of successfully registered cases was determined from the combined normal and fixed apex GR assessment results. The error in gray-value registration for both registration methods was determined from the position of one clearly defined calcification in the prostate gland (9 patients, 71 successful registrations). RESULTS: The percentage of successfully registered CBCT scans that were acquired with a collimated FOV was about 10% higher than for CBCT scans that were acquired with an uncollimated FOV. For CBCT scans that were acquired with a collimated FOV, the percentage of successfully registered cases improved from 65%, when only normal GR was applied, to 83% when the results of normal and fixed apex GR were combined. Gray-value registration mainly failed (or registrations were difficult to assess) because of streaks in the CBCT scans caused by moving gas pockets in the rectum during CBCT image acquisition (i.e., intrafraction motion). The error in gray-value registration along the left-right, craniocaudal, and anteroposterior axes was 1.0, 2.4, and 2.3 mm (1 SD) for normal GR, and 1.0, 2.0, and 1.7 mm (1 SD) for fixed apex GR. The systematic and random components of these SDs contributed approximately equally to these SDs, for both registration methods. CONCLUSIONS: The feasibility of automatic prostate localization on CBCT scans acquired on the treatment machine using an adaptation of the previously developed three-dimensional gray value registration algorithm, has been validated in this study. Collimating the FOV during CBCT image acquisition improved the CBCT image quality considerably. Artifacts in the CBCT images caused by large moving gas pockets during CBCT image acquisition were the main cause for unsuccessful registration. From this study, we can conclude that CBCT scans are suitable for online and offline position verification of the prostate, as long as the amount of nonstationary gas is limited. PMID- 16253773 TI - Influence of intravenous amifostine on xerostomia, tumor control, and survival after radiotherapy for head-and- neck cancer: 2-year follow-up of a prospective, randomized, phase III trial. AB - PURPOSE: To evaluate chronic xerostomia and tumor control 18 and 24 months after initial treatment with amifostine in a randomized controlled trial of patients with head-and-neck cancer; at 12 months after radiotherapy (RT), amifostine had been shown to reduce xerostomia without changing tumor control. METHODS AND MATERIALS: Adults with head-and-neck cancer who underwent once-daily RT for 5-7 weeks (total dose, 50-70 Gy) received either open-label amifostine (200 mg/m2 i.v.) 15-30 min before each fraction of radiation (n = 150) or RT alone (control; n = 153). RESULTS: Amifostine administration was associated with a reduced incidence of Grade > or =2 xerostomia over 2 years of follow-up (p = 0.002), an increase in the proportion of patients with meaningful (>0.1 g) unstimulated saliva production at 24 months (p = 0.011), and reduced mouth dryness scores on a patient benefit questionnaire at 24 months (p < 0.001). Locoregional control rate, progression-free survival, and overall survival were not significantly different between the amifostine group and the control group. CONCLUSIONS: Amifostine administration during head-and-neck RT reduces the severity and duration of xerostomia 2 years after treatment and does not seem to compromise locoregional control rates, progression-free survival, or overall survival. PMID- 16253774 TI - Economic consequence of local control with radiotherapy: cost analysis of internal mammary and medial supraclavicular lymph node radiotherapy in breast cancer. AB - PURPOSE: To investigate the financial implications of radiotherapy (RT) to the internal mammary and medial supraclavicular lymph node chain (IM-MS) in postoperative breast cancer. METHODS AND MATERIALS: A cost-effectiveness and cost utility analysis were performed, using Markov models, comparing the early and delayed costs and effects of IM-MS during a 20-year time span from a societal viewpoint. The outcome estimates were based on Level I evidence from postoperative RT literature and the cost estimates on the standard practice of the Leuven University Hospitals, with the RT costs derived from an activity-based costing program developed in the department. RESULTS: On the basis of the assumptions of the model and seen during a 20-year time span, primary treatment including IM-MS RT results in a cost savings (approximately 10,000) compared with a strategy without RT. Because IM-MS RT also results in better clinical effectiveness and greater quality of life, the treatment with IM-MS dominates the approach without IM-MS. Sensitivity analyses confirmed the robustness of these results in all tested circumstances. Although threshold values were found for the cost of IM-MS, the cost at relapse, and the quality of life after treatment, these were substantially different from the baseline estimates, indicating that it is very unlikely that omitting IM-MS would become superior. CONCLUSION: This ex-ante cost evaluation of IM-MS RT showed that the upfront costs of locoregional RT are easily compensated for by avoiding the costs of treating locoregional and distant relapse at a later stage. The cost-sparing effect of RT should, however, be evaluated for a sufficiently long time span and is most specifically found in tumors with a rather slow natural history and a multitude of available systemic treatments at relapse, such as breast cancer. PMID- 16253775 TI - Time-course of radiation-induced chromosomal aberrations in tumor patients after radiotherapy. AB - PURPOSE: Radiation-induced chromosome aberrations are routinely used in biologic dosimetry to monitor radiation exposure. Translocations are considered stable aberrations with time after exposure. This study was performed to determine the temporal persistence of radiation-induced translocations during a 36-month period in therapeutically irradiated testicular seminoma patients who underwent partial body exposure (>10% of bone marrow). METHODS AND MATERIALS: Chromosome analyses were carried out in peripheral lymphocytes of 11 patients with testicular seminoma (n = 9), germinoma (n = 1), or follicular non-Hodgkin's lymphoma (n = 1). All patients received radiotherapy with photons from a linear accelerator; in 1 case, additional electron beams were used. Doses ranged from 26 Gy (seminoma) to 45 Gy (non-Hodgkin's lymphoma). None of the patients received chemotherapy. From each patient, blood samples were taken during the 36 months after irradiation at defined points. Chromosomal aberrations were scored after fluorescence in situ hybridization painting of chromosomes 1, 4, and 12 in combination with a pancentromeric probe. RESULTS: For 9 patients (7 with testicular seminoma, 1 with germinoma, and 1 with non-Hodgkin's lymphoma), a significant temporal decline of translocations, with a mean decline rate of 4.4% +/- 0.4% monthly, could be detected. Two testicular seminoma patients showed no temporal decline of aberration frequencies. CONCLUSION: Most partial body irradiated patients (9 of 11) showed a significant temporal decline of translocation frequencies during a 36-month period. Thus, reciprocal translocations after partial body irradiation cannot be regarded as stable over time. The temporal decline of aberration frequencies has to be taken into account for retrospective dose estimations. PMID- 16253776 TI - Inhibition of phosphatidylinositol-3-kinase causes increased sensitivity to radiation through a PKB-dependent mechanism. AB - PURPOSE: To identify whether inhibition of phosphatidylinositol-3-kinase (PI3K) causes increased radiosensitivity through inhibition of protein kinase B (PKB), implicating PKB as an important therapeutic target in prostate cancer. METHODS AND MATERIALS: The prostate cancer cell line LNCaP was treated with the PI3K inhibitor LY294002, radiation, and combinations of the two therapies. Apoptosis and survival were measured by cell cycle analysis, Western blot analysis for cleaved poly (ADP-ribose) polymerase, and clonogenic survival. To test the hypothesis that inhibition of PKB is responsible for LY294002-induced radiosensitivity, LNCaP cells expressing a constitutively active form of PKB were used. RESULTS: The combination of PI3K inhibition and radiation caused an increase in apoptosis and a decrease in clonogenic survival when compared to either modality alone. The expression of constitutively activated PKB blocked apoptosis induced by combination of PI3K inhibition and radiation and prevented radiosensitization by LY294002. CONCLUSION: These data indicate that PI3K inhibition increases sensitivity of prostate cancer cell lines to ionizing radiation through inactivation of PKB. Therefore, PTEN mutations, which lead to PKB activation, may play an important role in the resistance of prostate cancer to radiation therapy. Targeted therapy against PKB could be beneficial in the management of prostate cancer patients. PMID- 16253777 TI - Clusterin as a therapeutic target for radiation sensitization in a lung cancer model. AB - PURPOSE: Clusterin plays important roles in cell survival and death. Inactivation of clusterin enhances the therapeutic efficacy of chemotherapy in lung cancer models. The purpose of this study was to determine whether inhibition of clusterin by an antisense-based investigative drug enhances radiation sensitization in a lung cancer model. METHODS AND MATERIALS: Cells were transfected with an antisense oligonucleotide (ASO) against clusterin (OGX-011). Apoptosis was determined by 7-aminoactinomycin D staining. Cell survival was examined by 3-(4, 5-methylthiazol-2-yl)-2, 5-diphenyl-tetrazolium bromide (MTT) and clonogenic assay. Xenograft model was used to demonstrate tumor growth and tumor blood flow. RESULTS: OGX-011 specifically attenuated the expression of secreted clusterin (prosurvival), with no apparent effect on the expression of nuclear clusterin (proapoptotic). Apoptosis was significantly increased when H460 lung cancer cells were treated with OGX-011 plus radiation. Inhibition of clusterin followed by radiation greatly decreased cell survival. H460 xenografts that were treated with OGX-011 plus radiotherapy demonstrated growth delay beyond 17 days. Doppler studies showed that tumor blood flow was compromised when mice bearing H460 xenografts were treated with OGX-011 and radiation. CONCLUSION: A combination of radiotherapy and OGX-011 improved control of tumor growth and vascular regression in the H460 lung cancer model. PMID- 16253778 TI - Enhanced efficiency in cell killing at the penetration depths around the Bragg peak of a radioactive 9C-ion beam. AB - PURPOSE: To evaluate the potential importance of radioactive 9C-ion beam in cancer radiotherapy. METHODS AND MATERIALS: Human salivary gland (HSG) cells were exposed to a double-radiation-source 9C beam at different depths around the Bragg peak. Cell survival fraction was determined by standard clonogenic assay. For comparison, the same experiment was conducted for a therapeutic 12C beam. To determine relative biologic effectiveness (RBE) values, HSG cells were also irradiated with 60Co gamma-rays of fractionation scheme as the reference. RESULTS: The 9C beam was more efficient in cell killing at the depths around its Bragg peak than was the 12C beam, which corresponded to the 9C-ion stopping region and where delayed low-energy particles were emitted. The RBE value at 50% survival level for the 9C beam varied from 1.38 to 4.23. Compared with the 12C beam, the RBE values for the 9C beam were always higher; an increase in RBE by a factor of up to 1.87 has been observed at the depths distal to the Bragg peak. CONCLUSION: The potential advantage of radioactive 9C-ion beam in cancer therapy has been revealed at low dose rate in comparison with a therapeutic 12C beam. This observation, however, remains to be investigated at therapeutic dose rates in the future. PMID- 16253779 TI - Changes in the tumor microenvironment during low-dose-rate permanent seed implantation iodine-125 brachytherapy. AB - PURPOSE: There is a lack of data regarding how the tumor microenvironment (e.g., perfusion and oxygen partial pressure [pO2]) changes in response to low-dose-rate (LDR) brachytherapy. This may be why some clinical issues remain unresolved, such as the appropriate use of adjuvant external beam radiation therapy (EBRT). The purpose of this work was to obtain some basic preclinical data on how the tumor microenvironment evolves in response to LDR brachytherapy. METHODS AND MATERIALS: In an experimental mouse tumor, pO2 (measured by electron paramagnetic resonance) and perfusion (measured by dynamic contrast-enhanced magnetic resonance imaging) were monitored as a function of time (0-6 days) and distance (0-2 mm and 2-4 mm) from an implanted 0.5 mCi iodine-125 brachytherapy seed. RESULTS: For most of the experiments, including controls, tumors remained hypoxic at all times. At distances of 2-4 mm from radioactive seeds ( approximately 1.5 Gy/day), however, there was an early, significant increase in pO2 within 24 h. The pO2 in that region remained elevated through Day 3. Additionally, the perfusion in that region was significantly higher than for controls starting at Day 3. CONCLUSION: It may be advantageous to give adjuvant EBRT shortly (approximately 1 to 2 days) after commencement of clinical LDR brachytherapy, when the pO2 in the spatial regions between seeds should be elevated. If chemotherapy is given adjuvantly, it may best be administered just a little later (approximately 3 or 4 days) after the start of LDR brachytherapy, when perfusion should be elevated. PMID- 16253780 TI - Caffeic acid phenethyl ester preferentially sensitizes CT26 colorectal adenocarcinoma to ionizing radiation without affecting bone marrow radioresponse. AB - PURPOSE: Caffeic acid phenethyl ester (CAPE), a component of propolis, was reported capable of depleting glutathione (GSH). We subsequently examined the radiosensitizing effect of CAPE and its toxicity. METHODS AND MATERIALS: The effects of CAPE on GSH level, GSH metabolism enzyme activities, NF-kappaB activity, and radiosensitivity in mouse CT26 colorectal adenocarcinoma cells were determined. BALB/c mouse with CT26 cells implantation was used as a syngeneic in vivo model for evaluation of treatment and toxicity end points. RESULTS: CAPE entered CT26 cells rapidly and depleted intracellular GSH in CT26 cells, but not in bone marrow cells. Pretreatment with nontoxic doses of CAPE significantly enhanced cell killing by ionizing radiation (IR) with sensitizer enhancement ratios up to 2.2. Pretreatment of CT26 cells with N-acetyl-L-cysteine reversed the GSH depletion activity and partially blocked the radiosensitizing effect of CAPE. CAPE treatment in CT26 cells increased glutathione peroxidase, decreased glutathione reductase, and did not affect glutathione S-transferase or gamma glutamyl transpeptidase activity. Radiation activated NF-kappaB was reversed by CAPE pretreatment. In vivo study revealed that pretreatment with CAPE before IR resulted in greater inhibition of tumor growth and prolongation of survival in comparison with IR alone. Pretreatment with CAPE neither affected body weights nor produced hepatic, renal, or hematopoietic toxicity. CONCLUSIONS: CAPE sensitizes CT26 colorectal adenocarcinoma to IR, which may be via depleting GSH and inhibiting NF-kappaB activity, without toxicity to bone marrow, liver, and kidney. PMID- 16253781 TI - Mapping of nodal disease in locally advanced prostate cancer: rethinking the clinical target volume for pelvic nodal irradiation based on vascular rather than bony anatomy. AB - PURPOSE: Toxicity from pelvic irradiation could be reduced if fields were limited to likely areas of nodal involvement rather than using the standard "four-field box." We employed a novel magnetic resonance lymphangiographic technique to highlight the likely sites of occult nodal metastasis from prostate cancer. METHODS AND MATERIALS: Eighteen prostate cancer patients with pathologically confirmed node-positive disease had a total of 69 pathologic nodes identifiable by lymphotropic nanoparticle-enhanced MRI and semiquantitative nodal analysis. Fourteen of these nodes were in the para-aortic region, and 55 were in the pelvis. The position of each of these malignant nodes was mapped to a common template based on its relation to skeletal or vascular anatomy. RESULTS: Relative to skeletal anatomy, nodes covered a diffuse volume from the mid lumbar spine to the superior pubic ramus and along the sacrum and pelvic side walls. In contrast, the nodal metastases mapped much more tightly relative to the large pelvic vessels. A proposed pelvic clinical target volume to encompass the region at greatest risk of containing occult nodal metastases would include a 2.0-cm radial expansion volume around the distal common iliac and proximal external and internal iliac vessels that would encompass 94.5% of the pelvic nodes at risk as defined by our node-positive prostate cancer patient cohort. CONCLUSIONS: Nodal metastases from prostate cancer are largely localized along the major pelvic vasculature. Defining nodal radiation treatment portals based on vascular rather than bony anatomy may allow for a significant decrease in normal pelvic tissue irradiation and its associated toxicities. PMID- 16253782 TI - 3D inverse treatment planning for the tandem and ovoid applicator in cervical cancer. AB - PURPOSE: Three-dimensional treatment planning systems and inverse planning optimization for brachytherapy are becoming commercially available. Guidelines for target delineation and dose constrictions have not been established using this new software. In this study we describe a method of target delineation for the tandem and ovoids applicator. We then compare inverse planning dose distributions with the traditional methods of prescribing dose. METHODS AND MATERIALS: Target and organ-at-risk volumes were defined using systematic guidelines on 15 patients treated in our department with high-dose-rate brachytherapy for cervical cancer using tandem and ovoids. High-dose-rate distributions were created according to three different dose optimization protocols: inverse planning simulated annealing (IPSA), point A, and point A with a normalization of 2 cc of the bladder receiving 80% of the dose (bladder-sparing method). An uniform cost function for dose constraints was applied to all IPSA generated plans, and no manual optimization was allowed for any planning method. RESULTS: Guidelines for target and structure-at-risk volumes, as well as dose constraint cost functions, were established. Dose-volume histogram analysis showed that the IPSA algorithm indicated no difference in tumor coverage compared with point A optimization while decreasing dose to the bladder and rectum. The IPSA algorithm provided better target volume coverage compared with bladder sparing method with equivalent doses to the bladder and rectum. CONCLUSION: This study uses a systematic approach for delineating target and organ-at-risk volumes and a uniform cost function for generating IPSA plans for cervical cancer using tandem and ovoids. Compared with conventional dose prescription methods, IPSA provides a consistent method of optimization that maintains or improves target coverage while decreasing dose to normal structures. Image-guided brachytherapy and inverse planning improve brachytherapy dosimetry. PMID- 16253783 TI - Is a "Puff to the Cuff" enough?: In regard to Alektiar et al. (Int J Radiat Oncol Biol Phys 2005;62: 111-117). PMID- 16253785 TI - In regard to Kao et al.: concomitant radiation therapy and paclitaxel for unresectable locally advanced breast cancer: results from two consecutive phase I/II trials (Int J Radiat Oncol Biol Phys 2005;61:1045-1053). PMID- 16253786 TI - Struggle for implementation of new strategies in intensive care medicine: anticoagulation, insulin, and lower tidal volumes. AB - The management of intensive care patients have changed dramatically in the last years: from merely supportive care, it has moved to evidence-based strategies that have been demonstrated to reduce mortality of the severely ill patients. Clinical research have brought numerous positive clinical trials offering intensive care physicians specific therapies to improve outcome of intensive care patients. Among them were the trials that tested the infusion of activated protein C in patients with severe sepsis, tight glycemic control in surgical intensive care patients, and use of lung protective mechanical ventilation by using small tidal volumes in patients with acute lung injury. Although results of these trials were sufficiently strong to, at least, consider implementation of these strategies in critical care medicine, published and yet unpublished reports show that there is significant struggle with implementation of these therapies. This manuscript focuses on the potential reasons that underlie this problem. PMID- 16253787 TI - Translating evidence into practice in the intensive care unit: the need for a systems-based approach. PMID- 16253788 TI - "Futile" care: do we provide it? Why? A semistructured, Canada-wide survey of intensive care unit doctors and nurses. AB - PURPOSE: To qualitatively explore the process of the provision of futile care in Canadian intensive care units (ICUs). MATERIALS AND METHODS: A mailed, semistructured survey was sent to medical and nursing unit directors of all Canadian ICUs, asking them to estimate the frequency of provision of futile care, when care becomes "futile," the reasons such care is provided, and the resources that are available to help make end-of-life decisions. Nurse/physician agreement was assessed by chi(2) analysis or Fisher exact test. Content analysis to identify common themes was carried out by 4 raters using a Delphi process. RESULTS: The response rate was 72%. The majority reported futile therapy had been provided in their ICU over the last year (nurses, "N"=95%, physicians, "P"=87%, P=.02). The most commonly stated reasons for providing futile care were family request (N=91%, P=91%, P=NS) and attending physician request (N=91%, P=87% P=NS). Physicians were cited to provide futile care because of prognostic uncertainty (N=73%, P=84%, P=.047) and legal pressures (N=84%, P=75%, P=NS). Comment review revealed 8 main reasons why futile care was provided, the most common of which were that "death was perceived as treatment failure," and poor provider-family communication. Few providers were aware of societal (N=26%, P=51%) or local (22%, all) guidelines relating to the provision of futile care, but of those who were aware, the majority found these useful (range, 73%-74%). Twenty-seven percent expressed the need for someone to discuss difficult ethical issues, such an individual with ethics training specifically assigned to the ICU. CONCLUSIONS: Caregivers voice the opinion that provision of futile care occurs, for multiple reasons, not the least of which is provider-driven. Nurses and physicians of Canadian ICUs perceive the need for increased availability of more ICU-directed and ethically trained resources to help them in providing end-of-life care. PMID- 16253789 TI - Barriers to communication regarding end-of-life care: perspectives of care providers. AB - OBJECTIVE: Communication regarding end-of-life care is frequently perceived as suboptimal, despite the intent of both health care providers and patients. We interviewed health care providers to determine their perspective regarding these barriers to communication. MATERIALS AND METHODS: Eleven focus groups with a total of 10 attending physicians, 24 residents, and 33 nurses were convened to explore barriers to end-of-life discussions on the Internal Medicine service at a 600-bed tertiary care hospital in Toronto, Canada. An interview schedule was designed to elicit information regarding the process of end-of-life discussions, barriers to these discussions, and possible interventions for limiting such barriers. Transcripts were qualitatively analyzed by 6 raters who independently identified "themes." Themes were refined using the Delphi technique and classified under broader "categories." RESULTS: Four main categories of barriers emerged, relating to (1) patients, (2) the health care system, (3) health care providers, and (4) the nature of this dialogue. Attending physicians and residents most frequently identified patient-related factors as barriers to discussions, followed by system, dialogue, and provider barriers (43%, 39%, 10%, and 8%, respectively, for attending physicians; 40%, 34%, 13%, and 13%, respectively, for residents). Nurses similarly identified patient-related and system barriers most frequently, but provider barriers were discussed more often than dialogue barriers (46%, 28%, 22%, and 4%, respectively). CONCLUSIONS: Attending physicians, residents, and nurses perceive the recipients of their care, and the system within which they provide this care, to be the major source of barriers to communication regarding end-of-life care. This finding may impact on the effectiveness of quality-improvement initiatives in end-of-life care. PMID- 16253790 TI - Ultrasound guidance during central venous catheterization: a survey of use by house staff physicians. AB - PURPOSE: The objective of this study is to explore the attitudes, knowledge, and behavior of house staff physicians regarding the use of ultrasound guidance (UG) during central venous catheter placement (CP). MATERIALS AND METHODS: A questionnaire-based study was used to survey house staff in multiple training programs at an academic tertiary care center. RESULTS: Fifteen percent of the respondents reported using UG on at least 60% of CP attempts. Those house staff physicians who agreed that UG is faster, easier, and more convenient than the landmark method reported more frequent use. Agreement that UG reduces the number of mechanical complications and placement failures was also associated with increased use. Respondents reporting the most CP experience were least likely to use UG. Most agreed that UG is useful in patients without good landmarks or when the landmark method failed, but these attitudes were not associated with the frequency of UG use. Most respondents were not aware of controlled trials demonstrating the benefits of UG. CONCLUSIONS: The use of UG during CP is infrequent despite the demonstrated benefits of this technology. Multiple knowledge and attitudinal barriers to the greater use of UG during CP must be addressed in the design of an effective UG implementation strategy. PMID- 16253791 TI - Prognosis factors and outcome of community-acquired pneumonia needing mechanical ventilation. AB - PURPOSE: To evaluate the variables associated with mortality of patients with community-acquired pneumonia who require mechanical ventilation and to determine the attributable morbidity and intensive care unit (ICU) mortality of community acquired pneumonia. MATERIAL AND METHODS: Retrospective cohort study carried out in 361 ICUs from 20 countries including 124 patients who required mechanical ventilation on the first day of admission to the hospital due to acute respiratory failure secondary to severe community-acquired pneumonia. To assess the factors associated with outcome, a forward stepwise logistic regression analysis was performed, and to determine the attributable mortality of community acquired pneumonia, a matched study design was used. RESULTS: We found 3 independent variables significantly associated with death in patients with community-acquired pneumonia requiring mechanical ventilation: simplified acute physiological score greater than 45 (odds ratio, 5.5 [95% confidence interval, 1.7-12.3]), shock (odds ratio, 5.7 [95% confidence interval, 1.7-10.1]), and acute renal failure (odds ratio, 3.0 [95% confidence interval, 1.1-4.0]). There was no statistically significant difference in ICU mortality among patients with or without community-acquired pneumonia (32% vs 35%; P=.59). CONCLUSIONS: Community-acquired pneumonia needing mechanical ventilation is not a disease associated with higher mortality. The main determinants of patient outcome were initial severity of illness and the development of shock and/or acute renal failure. PMID- 16253792 TI - Cost-effectiveness of immunoglobulin M-enriched immunoglobulin (Pentaglobin) in the treatment of severe sepsis and septic shock. AB - PURPOSE: To measure the cost-effectiveness of a specific polyclonal intravenous immune globulin preparation (Pentaglobin) in adult patients treated for severe sepsis and septic shock. MATERIALS AND METHODS: Effectiveness data from a meta analysis of 9 randomized trials (N=435) were used to populate a decision model to estimate the cost-effectiveness of Pentaglobin and its comparator standard therapy from the hospital perspective in Germany. PRIMARY OUTCOME: all-cause morality; secondary outcome: intensive care unit (ICU) length of stay. Benefit was expressed as lives saved (LS). Published cost data were applied to assess differences in ICU treatment costs. Cost-effectiveness was calculated as incremental cost per LS. RESULTS: Pentaglobin reduced the risk of mortality (P<.001) but had no effect on ICU length of stay. A baseline risk of mortality of 0.4434 (risk ratio=0.5652; absolute risk reduction=0.1928; number-needed-to treat=5.19) increased ICU treatment costs with Pentaglobin by 2,037 (22,711 vs 24,747) with a cost per LS of 10,565. Sensitivity analyses on baseline mortality risk (95% confidence interval 0.3293-0.5162) and risk ratio (95% confidence interval 0.4306-0.7420) yielded a cost per LS range of 5,715 to 28,443 with a 56.3% probability of cost-effectiveness of 12,000 or less. CONCLUSIONS: Pentaglobin is a promising adjuvant therapy both clinically and economically for treatment of adults with severe sepsis and septic shock. PMID- 16253793 TI - Putting the cart before the horse: a cautious look at the role of IgM-enriched polyclonal immunoglobulin in sepsis therapy. PMID- 16253794 TI - Readiness to change problematic drinking assessed in the emergency department as a predictor of change. AB - OBJECTIVES: The goal of this study was to determine if baseline readiness to change the drinking behavior (pre-contemplation, contemplation, preparation, and action stages) was predictive of change in drinking after unrelated emergency department (ED) visit and screening and interviewing for alcohol problems. METHODS: From August 1998 through December 2000, the Alcohol Use Disorders Identification Test (AUDIT) was administered to all consented ED patients aged 18 to 29 years. A brief motivational interviewing was provided to screen-positive patients (AUDIT score >5 of 40). Outcome at 3-month follow-up was measured as a decrease in the scores within the AUDIT domains of alcohol intake, harm, and dependency. RESULTS: Sixty percent of the screen-positive patients continued to drink at 3 months. Patients became more open to change their drinking behavior. Compared with patients in the pre-contemplation stage, those in the action stage were twice as likely to reduce their alcohol intake (OR, 2.24; 95% CI, 1.06 4.72), nearly 3 times as likely to reduce their alcohol-related harm behavior (OR, 2.80; 95% CI, 1.59-4.91), and almost 4 times more likely to decrease their dependency symptoms (OR, 3.59; 95% CI, 1.97-6.57). Compared with pre contemplation patients, those in the contemplation stage were nearly twice as likely to reduce their alcohol-related harm (OR, 1.85; 95% CI, 1.02-3.33) and those in the preparation stage were more than twice as likely to reduce their dependency symptoms (OR, 2.20, 95% CI, 1.13-4.27). CONCLUSIONS: Stages of change at baseline appeared to be significant predictors of change in alcohol intake, harm, and dependency symptoms among young adult ED patients. PMID- 16253795 TI - The reliability and validity of the therapeutic activity index. AB - PURPOSE: To assess the psychometric properties of the Simplified Therapeutic Intervention Scoring System (TISS 28) scale. MATERIALS AND METHODS: A prospective observational design was used. Patients were recruited from a medical-surgical intensive care unit (ICU) and 4 rehabilitation wards of 2 university-affiliated hospitals in Hong Kong. RESULTS: Data necessary for the calculation of the TISS 28, the Therapeutic Intervention Scoring System (TISS 76), and severity of illness scoring system (Simplified Acute Physiology Score [SAPS II]) were recorded for each patient during the first 24 hours after his/her admission to an ICU. A significant positive correlation was found between the TISS 76 and the TISS 28 scores as well as the TISS 28 and the SAPS II scores. There was a significant difference between the TISS 28 scores among ICU patients and patients in rehabilitation wards. A significant correlation was found between the TISS 28 scores of the first and second set of TISS 28 scores. CONCLUSIONS: Although the findings supported the validity and reliability of the TISS 28, there were limitations of the TISS 28 in measuring nursing workload in ICUs. Hence, continued amendment and validation of the TISS 28 on larger samples in different ICUs would be required so as to provide clinical nurses with a valid and reliable assessment of nursing workload. PMID- 16253796 TI - Health services research in critical care using administrative data. AB - The aim of health services research is to provide unbiased, scientific evidence to influence health services policy at all levels. Secondary analysis of administrative data can be employed for the purpose of evaluating questions relevant to health services research in the field of critical care. This article provides an overview of the topic and specifically reviews the key components to evaluating and performing research in critical care using administrative data, including how to evaluate the quality of administrative data itself, and also how to evaluate the quality of studies that employ administrative data. The article concludes with a step-wise approach to conducting studies of critical care based on administrative data. PMID- 16253797 TI - Medical education in critical care. AB - Patient safety is garnering increasing attention in our health care system, and with a possible trend toward increasing patient adverse events in academic hospitals (CMAJ. 2004;170:1678-86), medical education needs to reevaluate resident training. Within critical care, teaching of trainees in a manner that minimizes medical errors and ensures competency when managing critically ill patients is now a priority. This article addresses 4 areas that are receiving increasing attention in medical education and are relevant to training in the critical care environment: evidence-based medicine, communication, evaluation, and simulation. PMID- 16253798 TI - Incidence of acute respiratory distress syndrome and its relation to age. AB - PURPOSE: The incidence of acute respiratory distress syndrome (ARDS) was previously considered to be relatively low, at less than 10 cases per 100,000 inhabitants per year, but recent reports suggest a higher incidence, especially in elderly patients. The objective was to determine the incidence and mortality of ARDS in our setting, both overall and by age group. MATERIALS AND METHODS: We conducted a prospective, observational study of patients older than 14 years, admitted to the intensive care units of all hospitals in a province of southern Spain (Granada) during a 5-month period in 2001. American-European Consensus Conference criteria for ARDS were used. Patients were divided into 5 age groups, and the hospital mortality was recorded. RESULTS: During the study period, 61 Granada-residing patients developed ARDS criteria. This represents an overall incidence of 23 cases per 100,000 inhabitants per year in the province. The incidence of ARDS in the age groups of 15 to 29, 30 to 44, 45 to 59, 60 to 74, and older than 74 years was 4.6, 13.6, 21.6, 51, and 73.9 cases per 100,000 inhabitants per year, respectively. The overall hospital mortality rate was 66%. CONCLUSIONS: The incidence of ARDS is higher than reported a decade ago and is especially elevated in the elderly. The mortality remains high. PMID- 16253799 TI - Potassium sorbate reduces gastric colonization in patients receiving mechanical ventilation. AB - OBJECTIVES: Tube feeding might increase gastric burden of pathogenic bacteria and predispose patients to ventilator-associated pneumonia. We sought to determine whether a tube feeding formula acidified using potassium sorbate could reduce gastric burden of potentially pathogenic bacteria. DESIGN: Prospective, randomized, double-blind trial. SETTING: RML Specialty Hospital, a facility with expertise in weaning patients from prolonged mechanical ventilation. PATIENTS: Thirty patients recovering from prolonged mechanical ventilation. INTERVENTION: Patients were randomized to receive either a standard tube feeding formula (n=14) or a formula acidified using potassium sorbate to a pH of 4.25 (n=16). MEASUREMENTS AND RESULTS: Weekly quantitative cultures of gastric aspirates. The number of colony-forming units (CFUs) per patient was higher in the control than in the treatment group (53%+/-11% vs 9%+/-3.4%, threshold of >or=100,000 CFU/mL fluid, P=.003). The number of organisms isolated in each patient per week was higher among patients receiving standard tube feeding formula than among patients receiving acidified formula (0.91 +/- 0.20 vs 0.13 +/- 0.05 organisms per patient per week, threshold of >or=100,000 CFU/mL fluid, P=.0014). There was no difference in the incidence of gastrointestinal bleeding or ventilator-associated pneumonia between study groups. CONCLUSION: Tube feeding formula acidified using potassium sorbate was well tolerated and reduced gastric bacterial burden in patients recovering from prolonged mechanical ventilation. PMID- 16253801 TI - Inhaled nitric oxide therapy for persistent pulmonary hypertension of the newborn: when is it enough? PMID- 16253800 TI - Hypoxic respiratory failure in term newborns: clinical indicators for inhaled nitric oxide and extracorporeal membrane oxygenation therapy. AB - PURPOSE: The criteria for starting extracorporeal membrane oxygenation (ECMO) therapy in term newborn patients with hypoxemic respiratory failure consist of an oxygenation index (OI) of 25 or higher and alveolar-arterial oxygen (Aao(2)) gradient of more than 600 at sea level. In such conditions, inhaled nitric oxide (iNO) may improve oxygenation and reduce the need for ECMO therapy. We studied early changes in OI and Aao(2) gradients in response to iNO treatment that may indicate a need to continue iNO treatment or the necessity to start an ECMO therapy. MATERIALS AND METHODS: In this prospective study, we used 34 outborn neonatal patients that were referred to our pediatric critical care unit in a children's hospital for ECMO therapy with diagnosis of hypoxemic respiratory failure. In all patients, iNO therapy, starting at 80 ppm, was instituted either during transport or on arrival to hospital. Response to iNO was assessed after 1 hour, at which time, iNO concentration was reduced to 40 ppm, provided there was more than 20% improvement in either or both oxygenation indices. Patients who did not respond positively to continuous iNO therapy and met ECMO criteria were given ECMO therapy. RESULTS: Inhaled nitric oxide therapy alone was successful in 10 (29%) of 34 patients. Eighteen patients (53%) required ECMO therapy within the first 10 hours of iNO treatment (early ECMO therapy), whereas 6 other neonates (18%) became eligible for ECMO therapy after prolonged (2-4 days) iNO treatment (late ECMO therapy). No mortality occurred with any treatment. Within 4 hours after iNO therapy, patients who required early ECMO therapy had significantly higher OI and Aao(2) gradients than patients who were treated with iNO therapy alone (P<.01, analysis of variance followed by Tukey-Kramer multiple comparison test). Six of 34 patients (18%), categorized as late ECMO therapy, on the average, had initially higher levels of OI and mean airway pressure than neonates in iNO treatment and early ECMO therapy. CONCLUSION: Persisting levels of OI of more than 20 or Aao(2) gradients of more than 600 after 4 hours of iNO therapy could be indicative of an immediate need for ECMO therapy. PMID- 16253802 TI - Arterial thromboembolic complications in critically ill children. AB - PURPOSE: To evaluate incidence and characteristics of arterial thromboembolic complications in critically ill children. MATERIALS AND METHODS: Hospital records of all consecutive patients with arterial thromboembolic events (ATEs) occurring in the pediatric intensive care unit (PICU) from January 1997 to August 2001 were reviewed. Data collected included demographics and location, treatment modalities and outcome of ATEs. RESULTS: Fifty-four ATEs in 51 children (median age, 14 days) were identified, reflecting an incidence of 1.2% of all PICU patients. Arterial thromboembolic events were located in peripheral arteries in 52 (96%) cases and were associated with indwelling arterial catheters (n=26) or cardiac catheterization (n=26). The remaining 2 ATEs were located in the left ventricle and cerebral arteries, respectively. Therapy consisted of heparin (n=51), thrombolysis (n=22), oral anticoagulation (n=12), and aspirin (n=34). Complete resolution was noted in 33 (70%), partial resolution in 10 (21%), and no resolution in 4 (8.5%) cases. Bleeding complications occurred in 1 patient treated with heparin and in 12 (54%) of the 22 patients receiving thrombolytic therapy. CONCLUSIONS: Arterial thromboembolic events are frequent complications of PICU, particularly affecting neonates, and mostly associated with catheters. Studies to determine safe and effective prophylactic and treatment modalities of ATEs in children are required. PMID- 16253803 TI - Preferred stereoselective brain uptake of d-serine--a modulator of glutamatergic neurotransmission. AB - Although it has long been presumed that d-amino acids are uncommon in mammalians, substantial amounts of free d-serine have been detected in the mammalian brain. d Serine has been demonstrated to be an important modulator of glutamatergic neurotransmission and acts as an agonist at the strychnine-insensitive glycine site of N-methyl-d-aspartate receptors. The blood-to-brain transfer of d-serine is thought to be extremely low, and it is assumed that d-serine is generated by isomerization of l-serine in the brain. Stimulated by the observation of a preferred transport of the d-isomer of proline at the blood-brain barrier, we investigated the differential uptake of [3H]-d-serine and [3H]-l-serine in the rat brain 1 h after intravenous injection using quantitative autoradiography. Surprisingly, brain uptake of [3H]-d-serine was significantly higher than that of [3H]-l-serine, indicating a preferred transport of the d-enantiomer of serine at the blood-brain barrier. This finding indicates that exogenous d-serine may have a direct influence on glutamatergic neurotransmission and associated diseases. PMID- 16253804 TI - F-18 Polyethyleneglycol stilbenes as PET imaging agents targeting Abeta aggregates in the brain. AB - This paper describes a novel series of 18F-labeled polyethyleneglycol (PEG) stilbene derivatives as potential beta-amyloid (Abeta) plaque-specific imaging agents for positron emission tomography (PET). In these series of compounds, 18F is linked to the stilbene through a PEG chain, of which the number of ethoxy groups ranges from 2 to 5. The purpose of adding PEG groups is to lower the lipophilicity and improve bioavailability. The syntheses of the "cold" compounds and the 18F-labeled PEG stilbene derivatives are successfully achieved. All of the fluorinated stilbenes displayed high binding affinities in an assay using postmortem AD brain homogenates (K(i)=2.9-6.7 nM). Labeling was successfully performed by a substitution of the mesylate group of 10a-d by [18F]fluoride giving the target compounds [18F]12a-d (EOS, specific activity, 900-1500 Ci/mmol; radiochemical purity >99%). In vivo biodistribution of these novel (18)F ligands in normal mice exhibited excellent brain penetrations and rapid washouts after an intravenous injection (6.6-8.1 and 1.2-2.6 %dose/g at 2 and 60 min, respectively). Autoradiography of postmortem AD brain sections of [18F]12a-d confirmed the specific binding related to the presence of Abeta plaques. In addition, in vivo plaque labeling can be clearly demonstrated with these 18F labeled agents in transgenic mice (Tg2576), a useful animal model for Alzheimer's disease. In conclusion, the preliminary results strongly suggest these fluorinated PEG stilbene derivatives are suitable candidates as Abeta plaque imaging agents for studying patients with Alzheimer's disease. PMID- 16253805 TI - Imaging herpes viral thymidine kinase-1 reporter gene expression with a new 18F labeled probe: 2'-fluoro-2'-deoxy-5-[18F]fluoroethyl-1-beta-d-arabinofuranosyl uracil. AB - The preparation and radiolabeling of 2'-fluoro-2'-deoxy-1-beta-d-arabinofuranosyl 5-(2-fluoroethyl)-uracil (FFEAU) with 18F and its evaluation as a probe for imaging herpes simplex virus 1 thymidine kinase (HSV1-tk) gene expression are described. 2'-Fluoro-2'-deoxy-3',5'-di-O-benzoyl-1-beta-d-arabinofuranosyl-3-N benzoyl-5-(2-[18F]fluoroethyl)-uracil 12 was prepared by nucleophilic substitution of the corresponding tosyl 8 or trifluoroethanesulfonyl 9 derivative with n-Bu4N[18F]F. Base hydrolysis was used to remove the benzoyl protecting groups, followed by HPLC purification, to afford [18F]FFEAU 13. The trifluoroethanesulfonyl substrate 9 appears to be the better labeling precursor. Carrier n-Bu4NF was added to the labeling reaction, which resulted in specific activities of 40-70 Ci/mmol (estimated). Radiochemical purity averaged 94+/-4%. Although [18F]FFEAU was obtained in low radiochemical yield with 9 and further optimization of the radiosynthesis will be required, sufficient product was available for a series of in vitro and in vivo studies. [18F]FFEAU was directly compared with [3H]TdR in a series of in vitro accumulation studies involving a HSV1-tk stably transduced cell line, RG2TK+ and a nontransduced, wild-type RG2 cells. The initial in vitro and in vivo imaging studies are promising; FFEAU has in vitro accumulation and sensitivity characteristics similar to that previously reported for FIAU, but greater selectivity than FIAU due to lower uptake and retention in nontransduced cells and tissues. The animal imaging experiment showed low levels of radioactivity in the lungs, with little or no radioactivity seen in the heart, liver, spleen and intestines. PMID- 16253806 TI - Noninvasive imaging of transplanted living functional cells transfected with a reporter estrogen receptor gene. AB - The transplantation of functional cells such as dopaminergic cells into damaged tissue is now clinically ongoing, but at present the population of surviving cells at the transplantation site mostly cannot be noninvasively examined. To visualize surviving transplanted functional cells using a noninvasive method, we chose the estrogen receptor ligand binding domain (ERL) as a reporter molecule and 16alpha-[18F]-fluoro-17beta-estradiol (FES) for its ligand. We used a mouse embryonic stem (ES) cell line for recipient cells as a model. To obtain ES cells that constitutively or inducibly express ERL, we transfected two types of expression vectors into EB5 parental ES cell line using the lipofection method and obtained about 30 clones for each of the two types of transfectants. Then, to examine the expression level of ERL, we performed Western blotting analysis. Ligand uptake experiments were carried out using [3H]-estradiol with or without excessive unlabeled estradiol for control cells and ERL transfectants. Each selected clone was also used for in vivo positron emission tomography (PET) imaging studies involving FES in nude mice transplanted with control cells and ERL transfectants. In some of the clones transfected with the inducible-type ERL gene, protein was expressed much higher than in the controls. However, constitutive-type ERL gene-transfected ES cells showed no protein production in spite of their gene expression activity being considerably high. All clones also expressed equal levels of the Oct-3/4 gene, a marker of pluripotency, in comparison with the parental cells. Also, the specific uptake of [3H]-estradiol was over 30 times higher in inducer-treated ERL-expressing ES cells compared to untreated control cells. Finally, by performing dynamic PET imaging, we successfully visualized ERL-expressing teratomas using FES. PMID- 16253807 TI - Greater adenosine A(2A) receptor densities in cardiac and skeletal muscle in endurance-trained men: a [11C]TMSX PET study. AB - We examined the densities of adenosine A(2A) receptors in cardiac and skeletal muscles between untrained and endurance-trained subjects using positron emission tomography (PET) and [7-methyl-11C]-(E)-8-(3,4,5-trimethoxystyryl)-1,3,7 trimethylxanthine ([11C]TMSX), a newly developed radioligand for mapping adenosine A(2A) receptors. Five untrained and five endurance-trained subjects participated in this study. The density of adenosine A(2A) receptors was evaluated as the distribution volume of [11C]TMSX in cardiac and triceps brachii muscles in the resting state using PET. The distribution volume of [11C]TMSX in the myocardium was significantly greater than in the triceps brachii muscle in both groups. Further, distribution volumes [11C]TMSX in the trained subjects were significantly grater than those in untrained subjects (myocardium, 3.6+/-0.3 vs. 3.1+/-0.4 ml g(-1); triceps brachii muscle, 1.7+/-0.3 vs. 1.2+/-0.2 ml g(-1), respectively). These results indicate that the densities of adenosine A(2A) receptors in the cardiac and skeletal muscles are greater in the endurance trained men than in the untrained men. PMID- 16253808 TI - Synthesis and evaluation of [11C]FR194921 as a nonxanthine-type PET tracer for adenosine A1 receptors in the brain. AB - This report describes the synthesis of [11C]2-(1-methyl-4-piperidinyl)-6-(2 phenylpyrazolo[1,5-a]pyridin-3-yl)-3(2H)-pyridazinone ([11C]FR194921), a highly selective, nonxanthine-type adenosine A(1) receptor antagonist, used in brain imaging in rats and conscious monkeys as a potential novel PET tracer. [11C]FR194921 was successfully synthesized in 19 min after [11C]CH3I formation. The radiochemical yield was 38+/-3%; and radioactivity was 4.1+/-0.4 GBq, calculated from end of synthesis; radiochemical purity was higher than 99%; and the specific radioactivity was 25.0+/-8.1 GBq micromol(-1) (n=5). In a rat experiment, the distribution of [11C]FR194921 was higher in the hippocampus, striatum and cerebellum regions. This accumulation was significantly decreased by approximately 50% by pretreatment with 8-cyclopentyl-1,3-dipropylxanthine (DPCPX), an adenosine A1 receptor antagonist, which indicated specific binding of the radioligand to adenosine A1 receptors. In conscious monkey PET experiments, [11C]FR194921 accumulated in several regions of the brain, especially in the occipital cortex, thalamus and striatum. These results suggest that [11C]FR194921 can be used as an agent for imaging adenosine A1 receptors in vivo by positron emission tomography (PET). PMID- 16253809 TI - Distribution and pharmacokinetic analysis of angiostatin radioiodine labeled with high stability. AB - OBJECTIVE: Radiotracers of anticancer agents provide important information on its in vivo handling. Angiostatin (AST) is a promising anticancer drug with potent antiangiogenic effects, but reported AST radiotracers suffer from poor in vivo stability. In this study, we synthesized an AST probe radioiodinated via the Bolton-Hunter reagent (125I-BH-AST) and investigated its stability and biokinetics in mice. METHODS: 125I-BH-AST and conventional direct radioiodinated 125I-AST were evaluated for human endothelial cell binding characteristics. In vivo stability of the radiotracers was compared by biodistribution studies in normal ICR mice. Angiostatin pharmacokinetics was analyzed by serial blood sampling after intravenous injection of 125I-BH-AST with varying AST concentrations in mice. RESULTS: Both 125I-AST and 125I-BH-AST retained selective endothelial binding as demonstrated by dose-dependent inhibition by nonradiolabeled AST. 125I-BH-AST was substantially more stable in mice than 125I AST, with 28- and 7-fold lower 24-h thyroid and blood activities, respectively (15.5+/-1.5 vs. 430.9+/-32.2 and 0.1+/-0.0 vs. 0.8+/-0.0 %ID/g; both P<.005). Using (125)I-BH-AST, we found that 24-h AST accumulation was highest in the kidneys, followed by the liver and lungs. Kinetic analysis of 125I-BH-AST revealed AST to have linear pharmacokinetics with a T(1/2) of 5.8+/-2.6 h, volume of distribution (V(d)) of 6.8+/-1.3 ml and clearance of 0.8+/-0.1 ml/h. CONCLUSION: Radioiodine-labeled AST prepared by the BH method provides a radioprobe with superior stability and improved in vivo biokinetics that is useful for distribution and pharmacokinetic studies. PMID- 16253810 TI - Comparative uptakes and biodistributions of internalizing vs. noninternalizing copper-64 radioimmunoconjugates in cell and animal models of colon cancer. AB - Copper-64-labeled monoclonal antibodies (mAbs) have previously demonstrated unexpectedly effective tumor control in rodent models of cancer at relatively low tumor-absorbed radiation doses. This property has been associated with delivery platforms resulting in cellular internalization. The purpose of the present studies was to evaluate the in vitro internalization and in vivo distribution of a two-antibody model of 64Cu radioimmunotherapy (RIT) in the same cell and animal models of cancer. Biodistributions of an internalizing antibody, cBR96, and a noninternalizing antibody, cT84.66, labeled with 64Cu, were obtained in nude mice bearing LS174T colon carcinoma xenografts from 15 min to 48 h. The 64Cu-DOTA cBR96 conjugate demonstrated rapid tumor uptake, reaching 20.2% ID/g at 3 h and peaking at 35.4% ID/g by 24 h. Tumor accumulation of 64Cu-DOTA-cT84.66 was more gradual, 8.19% ID/g at 3 h and 43.8% ID/g by 24 h, but maximum uptake was not statistically different from 64Cu-DOTA-cBR96. Mouse xenograft dosimetry was estimated to be 1128 rad/mCi (304.9 mGy/MBq) for 64Cu-DOTA-cBR96 and 1409 rad/mCi (380.5 mGy/MBq) for 64Cu-DOTA-cT84.66. In LS174T cells, internalized radioactivity increased by a factor of 3.8 over 4 h for 64Cu-DOTA-cBR96, but remained unchanged 64Cu-DOTA-cT84.66. When normalized to uptake at 1 h, cellular efflux of 64Cu was essentially identical for both mAbs. The biodistributions and tumor dosimetry of these internalizing and noninternalizing radiolabeled mAbs were sufficiently similar for direct comparison of the therapeutic efficacies of low doses of 64Cu RIT agents in the same animal model of cancer. PMID- 16253811 TI - Biodistribution of 225Ra citrate in mice: retention of daughter radioisotopes in bone. AB - Alpha-particle-emitting radionuclides have potential for therapy of localized disease due to their high linear energy transformation and short pathlengths. Radiometals that home naturally to bone can be exploited for this purpose, and 223Ra (t(1/2)=11.4 days) recently has been studied for therapy of bone tumors in mice and rats. Actinium-225 (t(1/2)=10 days) is also an attractive radioisotope for endoradiotherapy. In a single decay of a 225Ac nucleus and its subsequent decay daughters, over 27 MeV ( approximately 90% of total energy) is released by sequential emission of four alpha particles, ranging in energy from 5.7 to 8.4 MeV. Although Ac3+ does not home naturally to bone, its parent radioisotope 225Ra (beta(-), t(1/2)=15 days) can be used as an in vivo source for 225Ac. Thus, injection of 225Ra takes advantage of the bone-homing properties of radium coupled with the significant amount of energy released from the 225Ac decay chain. Our data confirm that a large fraction of radium citrate injected intravenously into mice localizes rapidly in bone. Injected doses per gram (ID/g) for 225Ra range from 25% in skull to about 10% in sternum. Once deposited, the 225Ra remains in the bone with a biological half life of >40 days. Furthermore, >95% of the daughter radioisotope, 225Ac, is retained in the bone. However, a significant fraction of one of the daughter radioisotopes, 213Bi, is found in kidney. The biodistribution data indicate that 225Ra injection should be a powerful agent for killing cells associated with bone; however, the toxicity of this radioisotope which is similar to that of other alpha emitters limits the dose that can be tolerated. PMID- 16253812 TI - Whole-body radiation dosimetry of 2-[18F]Fluoro-A-85380 in human PET imaging studies. AB - 2-[18F]Fluoro-A-85380 (2-[18F]fluoro-3-(2(S)-azetidinylmethoxy)pyridine, 2 [18F]FA) is a recently developed PET radioligand for noninvasive imaging of nicotinic acetylcholine receptors. Previous radiation absorbed dose estimates for 2-[18F]FA were limited to evaluation of activity in only several critical organs. Here, we performed 2-[18F]FA radiation dosimetry studies on two healthy human volunteers to obtain data for all important body organs. Intravenous injection of 2.9 MBq/kg of 2-[18F]FA was followed by dynamic PET imaging. Regions of interest were placed over images of each organ to generate time-activity curves, from which we computed residence times. Radiation absorbed doses were calculated from the residence times using the MIRDOSE 3.0 program (version 3.0, ORISE, Oak Ridge, TN). The urinary bladder wall receives the highest radiation absorbed dose (0.153 mGy/MBq, 0.566 rad/mCi, for a 2.4-h voiding interval), followed by the liver (0.0496 mGy/MBq, 0.184 rad/mCi) and the kidneys (0.0470 mGy/MBq, 0.174 rad/mCi). The mean effective dose equivalent is estimated to be 0.0278 mSv/MBq (0.103 rem/mCi), indicating that radiation dosimetry associated with 2-[18F]FA is within acceptable limits. PMID- 16253813 TI - Techniques necessary for multiple tracer quantitative small-animal imaging studies. AB - INTRODUCTION: An increasing number and variety of studies on rodent models are being conducted using small-animal positron emission tomography scanners. We aimed to determine if animal handling techniques could be developed to perform routine animal imaging in a timely and efficient manner and with minimal effect on animal physiology. These techniques need to be reproducible in the same animal while maintaining hemodynamic and physiological stability. METHODS: The necessary techniques include (a) the use of inhalant anesthesia, (b) arterial and venous cannulation for multiple tracer administrations and blood sampling, (c) development of small-volume analytic columns and techniques and (d) measurement of the physiological environment during the imaging session. RESULTS: We provide an example of a cardiac imaging study using four radiotracers (15O-water, 1-[11C] acetate, 1-[11C]-palmitate and 1-[11C]-glucose) injected into normal rats. Plasma substrates, CO2 production and total metabolites were measured. The animals remained anesthetized over the entire imaging session, and their physiological state was maintained. CONCLUSION: The intrastudy stability of the physiological measurements and substrate levels and interstudy reproducibility of the measurements are reported. PMID- 16253814 TI - An improved (99m)Tc-aprotinin kit formulation: quality control analysis of radiotracer stability and cold kit shelf life. AB - (99m)Tc-aprotinin scintigraphy has been demonstrated to be a useful noninvasive imaging technique for amyloid deposits located in extraabdominal regions of patients. The aim of this study was to develop an improved aprotinin cold kit formulation, to validate the kit for long-term stability, as well as to assess the radiotracer stability by novel quality control methods. The aprotinin cold kit formulation of Trasylol, pyrophosphate (PYP)-chelated stannous reductant and an alkaline buffer, was dispensed into nitrogen-filled vials and aliquots frozen at -20 degrees C. After 0, 1, 2, 3 and 6 months of storage, three samples were reconstituted with 750-850 MBq of (99m)Tc-pertechnetate, followed by quality control analyses by paper chromatography methods at 25, 85 and 265 min postreconstitution (pr). Cation-exchange cartridge quality control methods were also investigated. The cold kits proved to be stable to long-term storage for up to 6 months, and the radiotracer was stable for at least 4 h pr. (99m)Tc aprotinin was formed at greater than 95% efficiency at all time points tested with (99m)TcO2 present as the major impurity (1-4%) and (99m)Tc-pertechnetate and (99m)Tc-PYP present in trace amounts. An alternative, rapid, safe and reliable method was found in Oasis MCX-BSA-treated cartridges using saline as the eluting solution to assay for (99m)Tc-aprotinin. PMID- 16253815 TI - Preparation of high specific activity (86)Y using a small biomedical cyclotron. AB - 86Y is an attractive PET radionuclide due to its intermediate half-life. (86)Y was produced via the 86Sr(p,n)86Y nuclear reaction. Enriched SrCO3 or SrO was irradiated with 2-6 microA of beam current for <4 h on a CS-15 cyclotron. It was shown that the SrO target could withstand at least 6 microA of beam current, a significant improvement over a maximum of 2 microA on the SrCO3 target. Average yields of 4.5 mCi/microA.h were achieved with SrO, which represent 71% of the theoretical yield, compared to 2.3 mCi/microA.h with SrCO3. The radioisotopic contaminants were (86m)Y (220%), 87Y (0.27%), (87m)Y (0.43%) and 88Y (0.024%). 86Y was isolated in an electrochemical cell consisting of three Pt electrodes. The solution was electrolyzed at 2000 mA (40 min) using two Pt plate electrodes. A second electrolysis (230 mA for 20 min) was performed using one Pt plate and a Pt wire. On average, 97.1% of the 86Y was recollected on the Pt wire after a second electrolysis. The (86)Y was collected from the Pt wire using 2.8 M HNO3/EtOH (3:1). After evaporation, 86Y was reconstituted in 100 microl of 0.1 M HCl. Target materials were recovered as SrCO3 and then converted to SrO by thermal decomposition at 1150 degrees C. Specific activity of 86Y was determined to be 29+/-19 mCi/microg via titration of 86Y(OAc)3 with DOTA or DTPA. We have established techniques for the routine, economical production of high purity, high specific activity 86Y on a small biomedical cyclotron that are translatable to other institutions. PMID- 16253816 TI - Fully automated synthesis of [18F]fluoromisonidazole using a conventional [18F]FDG module. AB - We developed a new fully automated method for the synthesis of [18F]fluoromisonidazole ([18F]FMISO) by modifying a commercial FDG synthesizer and its disposable fluid pathway. A three-step procedure was used to prepare the tosylate precursor, 1-(2'-nitro-1'-imidazolyl)-2-O-tetrahydrofuranyl-3-O toluenesulfonylpropanediol. Using glycerol as the starting material, the precursor was synthesized with a yield of 21%. The optimal labeling conditions for the automated synthesis of [18F]FMISO was 10 mg of precursor in acetonitrile (2 ml heated at 105 degrees C for 360 s, followed by heating at 75 degrees C for 280 s and hydrolysis with 1 N HCl at 105 degrees C for 300 s. Using 3.7 GBq of [18F]F- as a starting activity, [18F]FMISO was obtained with high end-of synthesis (EOS) radiochemical yields of 58.5+/-3.5% for 60.0+/-5.2 min with high performance liquid chromatography (HPLC) purification. When solid-phase purification steps were added, the EOS radiochemical yields were 54.5+/-2.8% (337+/-25 GBq/micromol) for 70.0+/-3.8 min (n=10 for each group, decay corrected). With a high starting radioactivity of 37.0 GBq, we obtained radiochemical yields of 54.4+/-2.9% and 52.8+/-4.2%, respectively (n=3). The solid-phase purification removed unreacted [18F]fluoride and polar impurities before the HPLC procedure. Long-term tests showed a good stability of 98.2+/ 1.5%. This new automated synthesis procedure combines high and reproducible yields with the advantage of using a disposable cassette system. PMID- 16253817 TI - Improved quality control of [18F]FDG by HPLC with UV detection. AB - A conventional high-performance liquid chromatographic (HPLC) method for the analysis of 2-fluoro-2-deoxy-d-glucose (FDG) and 2-deoxy-2-chloro-d-glucose (ClDG) in [18F]FDG preparations is described. This method was based on a postcolumn derivatization with 2-cyanoacetamide (2-CA) and UV detection. FDG and ClDG were separated on a normal-phase column using acetonitrile/water as the mobile phase. The eluate was mixed with 2-CA in sodium borate buffer solution at the outlet of a PTFE coil (10 m x 0.5 mm id) from the column, and the reaction was carried out at 100 degrees C during the passage through the coil. The UV absorbance of the resultant product was monitored at 276 nm. Under optimum conditions, the detection limits [signal-to-noise (S/N) ratio=3] for FDG and ClDG were 0.31 and 0.17 microg/ml for a 20-microl injection volume, respectively, and the linearity ranges were 0.5-100 microg/ml for both compounds. The intra- and interday reproducibilities were better than 2.2% [relative standard deviation (R.S.D.)]. This HPLC separation procedure is also useful for determining the radiochemical purity of [18F]FDG preparations since it allows the analysis of 2 [18F]fluoro-1,3,4,6-tetra-O-acetyl-d-glucose ([18F]TAG), partially hydrolyzed [18F]TAG and [18F]F-. This method can be used at many positron emission tomography (PET) facilities since it does not require an expensive, sophisticated electrochemical detector. PMID- 16253818 TI - Adjuvant therapies for non-small cell lung cancer: introduction. PMID- 16253819 TI - Molecular staging and the selection of therapy for non-small cell lung cancer. AB - The stage-specific selection of therapy is the standard for patients with non small cell lung cancer. Investigation of the molecular biology of lung cancer has provided pathways and targets that may be used to improve the efficacy of therapy and improve the survival for patients with lung cancer. PMID- 16253820 TI - Surgical strategies and outcomes after induction therapy for non-small cell lung cancer. AB - Surgery as the sole therapy for locally advanced non-small cell lung cancer (NSCLC) is usually not curative. Adjuvant chemotherapy has been evaluated by several randomized Phase III trials and found to confer a survival benefit over surgery alone for stage IB-IIIA NSCLC. Induction therapy applies a cytoreductive and systemic therapy before definitive locoregional therapy. Theoretical advantages include improved diffusion of chemotherapy agents into the tumor, improved compliance, and a higher complete resection rate. Results from multiple Phase II and III studies have been encouraging, but the role of surgery after induction therapy remains inconclusively defined. Randomized trials are underway to better define the role of induction therapy, and enrollment of patients into such trials should be encouraged. PMID- 16253821 TI - Controversial issues regarding the use of induction chemotherapy for lung cancer. AB - Induction chemotherapy has been proven to improve survival in patients with Stage IIIA non-small cell lung cancer, and is under investigation for early stage disease. Controversy still exists regarding the choice of chemotherapy regimens, patient selection, and inclusion of radiation therapy. PMID- 16253822 TI - Adjuvant chemotherapy for non-small cell lung cancer. AB - The survival after complete resection for non-small cell lung cancer (NSCLC) is unsatisfactory. Until recently, the use of adjuvant therapy after resection for early stage disease has not been proven to improve survival. However, the efficacy of adjuvant therapy has been demonstrated in phase III prospective randomized trials. The appropriate use of adjuvant therapy, including biologic therapy, is currently under investigation. PMID- 16253823 TI - Targeted therapy for non-small cell lung cancer. AB - The overall survival for the treatment of lung cancer patients is less than 15%, despite advances in chemotherapy, radiation therapy, and surgery, due to the inability to control metastatic disease. Over the past three decades, the genetics of lung cancer has been progressively delineated. Small molecule drugs or monoclonal antibodies have been developed that target and inactivate specific cancer-related proteins, such as growth factor receptors or their kinases. This article will review the therapeutic implications of molecular changes associated with non-small cell lung cancer and the status of targeted therapies in its treatment. PMID- 16253824 TI - Gene therapy for lung cancer. AB - Over the past three decades, the molecular biology of lung cancer has been progressively delineated. Concurrently, gene therapy techniques have been developed that allow targeting or replacement of dysfunctional genes in cancer cells, such as activated tumor-promoting oncogenes, inactivated tumor suppressing, or apoptosis-promoting genes. This article will review the therapeutic implications of molecular changes associated with non-small cell lung cancer and the status of gene therapy. PMID- 16253825 TI - Contemporary management of aortic dissection: introduction. PMID- 16253826 TI - Diagnostic imaging for aortic dissection. AB - Diagnostic imaging for aortic dissection has dramatically changed in recent years. Previously, imaging consisted of conventional X-ray radiography, followed by invasive catheter angiography. Now imaging of dissection is performed primarily with multidetector CT, and to a lesser extent, with ultrasound and MRI. Catheter angiography is used primarily as a means of treating complications. Which modality to choose depends on patient factors, physician preference, and differences in availability of state-of-the-art equipment. All three modalities are highly accurate in experienced hands and have revolutionized the detection and evaluation of this condition. PMID- 16253827 TI - Indications, timing, and prognosis of operative repair of aortic dissections. AB - Since the first description of aortic dissection in the 1700s, the understanding and treatment of this catastrophic disease has evolved. Aortic dissections are identified as a tear in the aortic intima and inner layer of the media that allows for blood flow within the aortic wall. The area of the vessel involved determines its classification. The classification, in turn, helps to predict outcomes, which allows for appropriate treatment planning. The goal of this article is to outline the operative indications and timing for Stanford type A and type B dissections, based on prior reported data and our own clinical experience with 176 patients treated surgically at the Yale Center for Thoracic Aortic Disease. With this data we will revisit the importance of looking at each patient individually to devise an appropriate operative plan, with the knowledge that treatment for type A dissections is operative and treatment for type B dissections is medical unless patients present with actual or impending rupture, malperfusion, or failure of medical management. PMID- 16253828 TI - Anesthesia and neurocerebral monitoring for aortic dissection. AB - Patients presenting to the operating room for repair of aortic dissection are challenging in all aspects of their care. Without exception, they require a multidisciplinary team approach. This article will review some of the specific challenges faced by anesthesiologists and neurologists when confronted with such a diagnosis. Specifically, we will discuss the myriad anesthetic issues that present in the preoperative stage and continue into the postoperative period. Neurologic complications during dissection repair result in increased morbidity and mortality. A variety of neurophysiologic monitoring techniques exist that may reduce this risk and will be discussed in detail. Finally, we will present some "controversies in care," emphasizing that our respective fields continue to grow, learn, and improve what information we have on the morbidity and mortality of aortic dissection. PMID- 16253829 TI - Operative management of ascending aortic dissections. AB - Surgical management has been the established standard of care for acute ascending aortic dissection (Stanford type A or Debakey type I or II) with a worldwide average operative mortality rate of 23% compared with a 1-month mortality rate of 60% with medical management alone. Improving an institution's operative results depends on the establishment of a comprehensive and specialized team integrating their expertise into a protocol-driven system of care delivery specific for aortic dissections. The operative strategies and techniques for repairing aortic dissections should also be standardized within an institution to optimize outcomes. The primary goals of operative repair of ascending aortic dissection are to restore aortic valvular competency and systemic perfusion, to obliterate false-lumen blood flow, and to prevent rupture, myocardial infarction, stroke, and death. The goals, strategies, and expected outcomes for the operative management of acute ascending aortic dissection are presented. PMID- 16253830 TI - Acute descending aortic dissections: management of visceral, spinal cord, and extremity malperfusion. AB - Acute descending aortic dissection is considered the most catastrophic event affecting the aorta and occurs two to three times more often than rupture of abdominal aortic aneurysms. The therapeutic aim in treating acute dissection is not only directed at the prevention of aneurysmal development and rupture but also to prevent and treat complications such as malperfusion syndrome. According to Lauterbach and coworkers patients with symptomatic malperfusion syndromes have a 51% mortality rate compared with a 29% mortality rate in patients who do not. The surgical in-hospital mortality rate in patients with mesenteric or peripheral vascular ischemic complications may be as high as 89%. Despite an improvement in diagnosing dissections and malperfusion syndromes, and despite improved operative techniques and a better understanding of the significance of perioperative care, the surgical mortality rate can be as high as 50%. Endovascular techniques are constantly evolving that provide an alternative to open procedures. The goal of this article was to review the pathogenesis of malperfusion syndromes in aortic dissection, discuss the current modalities to treat malperfusion of the spinal cord, viscera, and extremities, and examine the results of the treatments used today. PMID- 16253831 TI - Chronic descending aortic dissections. AB - Aortic dissection involving the descending aorta has a predictable and often debilitating course of progressive dilatation that occurs once patients survive the acute phase of the disease. Important factors that impact the rate and degree of dilatation include the persistence of a false lumen channel (and the amount of thrombus), the control of hypertension, and the presence of an underlying connective tissue disorder. The mainstay of management of chronic descending aortic dissections is antihypertensive therapy including beta-blockade until the dissected aorta becomes significantly aneurysmal. Surgical management is recommended at that point. Multiple advances have been made that have improved the results of operative repair of aneurysmal descending aortic dissections including circulation management methodologies, CSF drainage, neurocerebral monitoring, and more recently, endovascular therapies. The presentation, diagnosis and management of chronic descending aortic dissections are discussed. PMID- 16253832 TI - Endovascular repair of descending aortic dissections. AB - Endoluminal aortic stent-grafts have revolutionized the contemporary management of abdominal aortic aneurysms. The application of this treatment option for aortic dissections has been hampered by the difficult anatomic constraints frequently encountered with both acute and chronic forms of the disease. Although the data are sparse, it is rapidly growing as technologic advances are made in the design and delivery of various stent-grafts. Reviewed is our current understanding of the pathophysiology of descending thoracic aortic dissections, available stent-graft designs, techniques of implantation, and alternatives for complex anatomy. The section concludes with an overview of published data on stent-graft repair of both acute and chronic aortic dissections. PMID- 16253833 TI - Heritable disorders predisposing to aortic dissection. AB - Heritable disorders of connective tissue often predispose patients to aortic pathology and in particular aortic dissection. The Marfan syndrome, vascular Ehlers-Danlos syndrome, familial forms of thoracic aortic aneurysms or aortic dissection, and bicuspid aortic valve are all examples of heritable disorders that have associated defects affecting the integrity of the aortic wall, posing a risk of both aneurysmal dilation and dissection. The purpose of this review was to outline the phenotypes of the heritable syndromes that predispose to aortic dissection, present a guideline to their management and surveillance, and to offer insight into some of the surgical pitfalls that occur when repairing ascending aortic dissections in these types of patients. PMID- 16253835 TI - Anatomy of the eyelids. AB - The variety and complexity of periorbital surgical procedures continue to increase. Successful completion of reconstructive and esthetic ocular procedures requires an exacting knowledge of the relevant anatomy. Interestingly, the definition of ocular and periorbital anatomy continues to evolve, including more recent descriptions of the orbitomalar ligament and a new understanding of eyelid lymphatics. This article presents the most recent descriptions of periorbital anatomy. PMID- 16253836 TI - The beautiful eye. AB - Improved esthetic results in the periorbital region can be accomplished with a thorough knowledge of the features of the "beautiful eye." The information provided in this article, culled from the literature and years of clinical experience, should allow the reader to develop an improved esthetic sensibility as applied to the periorbital region. Deviations from the "ideal" can be identified properly and addressed appropriately, leading to improved esthetic outcomes. PMID- 16253837 TI - Evolution of eyelid surgery. AB - Cosmetic rejuvenation of the periorbital area has been an area of continual advancement in surgical knowledge over the past century. It is anticipated that some new currently described advances will be incorporated into continuing practice patterns, with advances in nonsurgical rejuvenation and surgical techniques. PMID- 16253838 TI - Upper lid blepharoplasty. AB - Upper lid blepharoplasty is the most commonly performed operation in patients seeking facial rejuvenation. When evaluating the patient, attention should be focused on the possibility of concomitant eyelid or eyebrow ptosis. Meticulous planning and technique are required to achieve high levels of patient satisfaction. PMID- 16253839 TI - Upper blepharoplasty in the Asian eyelid. AB - Upper eyelid blepharoplasty is a major component of cosmetic surgery in the Asian face. The complicated and variable anatomy of the Asian upper eyelid is reviewed, stressing the major differences between Asian and Caucasian eyes. The authors' preferred surgery for the creation of a double eyelid is discussed, including the preoperative assessment, intraoperative maneuvers, and laser techniques. The article concludes with a discussion of the complications specifically related to Asian blepharoplasty and strategies to avoid them. PMID- 16253840 TI - Ptosis repair for the cosmetic surgeon. AB - The cosmetic surgeon must be familiar with the evaluation and management of the ptotic eyelid. A small amount of unrecognized preoperative eyelid ptosis can seem obtrusive if it is unmasked after blepharoplasty. As such, ptosis repair is an important adjunctive procedure in cosmetic eyelid surgery. This article describes the basics behind the two most common procedures to correct eyelid ptosis. Cosmetic eyelid surgeons should familiarize themselves with these procedures or work in conjunction with an ophthalmic plastic surgeon who can assist in addressing this common eyelid malposition. PMID- 16253841 TI - Lower lid blepharoplasty and canthal surgery. AB - Lower lid blepharoplasty is an effective procedure to address aging changes of the eyes and periorbital regions. Success requires a thorough understanding of the relevant anatomy, proper patient selection, and meticulous surgical technique. When performed skillfully, the transcutaneous and transconjunctival approaches can offer the patient excellent results with a low risk of complications. PMID- 16253842 TI - Lower blepharoplasty: transconjunctival fat repositioning. AB - Traditionally, lower eyelid herniated fat is removed, which may cause a sunken or hollow lid appearance, especially in patients with a tear-trough deformity (nasojugal groove). Lower eyelid transconjunctival fat repositioning, defined as the subperiosteal repositioning of the medial and central lower eyelid herniated orbital fat into the nasojugal fold, may prevent the surgical hollow lower eyelid appearance while treating the herniated fat. Fat repositioning may be combined with an endoscopic subperiosteal midface-lift, transcutaneous skin pinch, and transconjunctival orbicularis oculi excision. This technique offers a powerful tool in the surgical armamentarium of the facial plastic surgeon. PMID- 16253843 TI - Revisional eyelid surgery: treatment of severe postblepharoplasty lower eyelid retraction. AB - A common and challenging complication of lower blepharoplasty is severe lower eyelid retraction. This complication is esthetically displeasing and can result in ocular discomfort, blurred vision, and exposure keratopathy. The hard palate graft and cheek-lift procedures can eliminate the lower lid retraction without creating disfiguring facial scars. The authors discuss their approach to the treatment of severe lower eyelid retraction after cosmetic blepharoplasty. PMID- 16253844 TI - Periorbital ablative and nonablative resurfacing. AB - Ablative resurfacing using laser systems for facial rejuvenation, especially in the periorbital areas, has effectively replaced chemical peels. Refined methods yield outstanding efficacy with significantly fewer side effects as compared with the prototypical lasers of decades past. Despite these technologic refinements, the trend toward minimally invasive rejuvenation techniques has placed a substantial emphasis on the many nonablative modalities available to the practitioner. Although the clinical efficacy to date has been less with the nonablative devices, the lack of substantial morbidity has made the devices marketable to a large population of patients despite their modest results at best and highly unpredictable results at worst. Numerous options, ablative and nonablative, are now available for the patient desiring improvement in periorbital rhytids. PMID- 16253845 TI - Primary and adjunctive uses of botulinum toxin type A in the periorbital region. AB - Chemodenervation with botulinum toxin has become an integral part of the facial plastic armamentarium. Although eyebrow and eyelid cosmetic deformities and asymmetries have traditionally been treated by surgical intervention, Botox can now be incorporated effectively into the treatment plan. When the surgeon has a complete understanding of facial anatomy and muscular interactions, Botox can be used as a primary treatment of the periorbital region or as an adjunct to surgical procedures of the periorbital region. PMID- 16253846 TI - Proceedings from the 2004 International Workshop on Functional Pulmonary Imaging, Philadelphia, Pennsylvania, USA. PMID- 16253847 TI - Current status of functional pulmonary imaging. PMID- 16253848 TI - Hyperpolarized 3He MRI in asthma measurements of regional ventilation following allergic sensitization and challenge in mice--preliminary results. AB - RATIONALE AND OBJECTIVES: Quantitative regional measurement of physiological parameters of lung may improve both early detection of asthma and its response to treatment by elucidating the characteristics of airway obstruction. Recent emergence of hyperpolarized helium-3 magnetic resonance imaging as a sensitive pulmonary imaging tool has shown great potential in capturing important structural and functional aspects of normal and diseased lungs. The objective of this study was to investigate regional ventilation changes in the mouse lung following allergen sensitization and challenge. MATERIALS AND METHODS: A murine model of allergic airway inflammation was created in mice following allergen challenge using Af and IgE-mediated asthma. The creation of model was verified using pulmonary function test and histology. Regional fractional ventilation was then measured in the animals using hyperpolarized 3He MRI on a pixel-by-pixel basis with a planar resolution of 0.24 mm. The sensitized and healthy animals were then compared statistically to assess the potential sensitivity of this technique in detection of such pulmonary abnormalities. RESULTS: In this work, we have demonstrated for the first time the quantitative measurement of regional ventilation in normal and asthmatic mice. Results of this study show significant changes in regional ventilation in murine model of allergic airway sensitization compared with that in normal control animals. CONCLUSION: Further development of this technique can potentially serve as a quantitative marker to investigate the physiology of allergen-induced airway hyperresponsiveness and to assist in disease treatment and prevention. PMID- 16253850 TI - Detection of age-dependent changes in healthy adult lungs with diffusion-weighted 3He MRI. AB - RATIONALE AND OBJECTIVES: To investigate changes in lung microstructure in healthy adult subjects with no smoking history using diffusion-weighted 3He MRI. MATERIALS AND METHODS: Diffusion magnetic resonance imaging using hyperpolarized helium 3 (3He) was applied to healthy volunteers to explore the dependence of lung microstructural changes with age, reflected by changes in the apparent diffusion coefficient (ADC) of 3He in lung air spaces. Data from three sites (University of Virginia (UVa), N = 25; University of Wisconsin (UW), N = 8; University of Nottingham (UN), N = 11) were combined in pooled analysis, including a total of N = 44 subjects (age range, 18-69 years; average age, 41.7 +/- 16.7 years). RESULTS: ADC was found to depend on age at all three sites (UW, R = +0.95, P = .0003; UVa, R = +0.74, P < .0001; UN, R = +0.96, P < .0001). Increases in mean ADCs with age appeared similar across sites (UW, +0.0017 cm2 s( 1) y(-1); UVa, +0.0015 cm2 s(-1) y(-1); pooled, +0.0015 cm2 s(-1) y(-1); P = .71). In a regional analysis performed on UW data, the increase in ADC affected all regions of the lung, but the apical and middle regions showed a greater increase compared with the base of the lung. CONCLUSION: Results suggest the observed age dependence of the ADC may be caused by changes in lung microstructure that increase alveolar volume during the aging process. PMID- 16253849 TI - Static versus prospective gated non-breath hold volumetric MDCT imaging of the lungs. AB - RATIONALE AND OBJECTIVES: The study's aim is to establish lung-imaging methods that provide for the ability to image the lung under dynamic non-breath hold conditions while providing "virtual breath hold" quantifiable volumetric image data sets. Static breath hold images are used as the gold standard for evaluating these virtual breath hold images in both a phantom and sheep. MATERIALS AND METHODS: Axial methods for gating image acquisition to multiple points in the respiratory cycle interleaved with incremental table stepping during multidetector-row computed tomographic (MDCT) scanning were developed. Data sets are generated over multiple breaths, providing volume images representative of multiple points within a respiratory cycle. To determine the reproducibility and accuracy of the methods, six anesthetized sheep were studied by means of MDCT in nongated and airway-pressure (P(awy))-gated modes in which P(awy) was 0, 7, and 15 cm H2O. RESULTS: No significant differences were found between coefficients of variation in air volume measured from repeated static scans (1.74% +/- 1.78%), gated scans: inspiratory (1.2% +/- 0.44%) or expiratory gated (1.39% +/- 0.98%), or between static (1.74% +/- 1.78%) and gated (1.39% +/- 0.98%) scanning at similar P(awy) (P > .1). Measured air volumes were larger from static versus gated scans by 5.85% +/- 3.77% at 7 cm H2O and 4.45% +/- 3.6% at 15 cm H2O of P(awy) (P < .05), consistent with hysteresis. Differences between air volumes at 7 and 15 cm H2O measured from either static or gated scans or that delivered by a super syringe were insignificant (P < .05). Visual accuracy of three-dimensional anatomic geometry was achieved, and landmark certainty was within 1 mm across respiratory cycles. CONCLUSIONS: A method has been shown that provides for accurate gating to respiratory signals during axial scanning. High-resolution volumetric image data sets are achievable while the scanned subject is breathing. Images are quantitatively similar to breath hold images, with differences likely explained by known pressure-volume hysteresis effects. PMID- 16253851 TI - Molecular imaging of the lungs. AB - An emerging suite of new imaging techniques offer the ability to monitor and quantify molecular and cellular processes in the lungs noninvasively. These techniques take advantage of dramatic advances in both imaging technology as well as molecular and cell biology. Molecular imaging is being used with increasing regularity in research protocols, and forms of molecular imaging have found their way into the patient care setting (eg, positron emission tomography imaging in cancer). Such techniques will afford the basic scientist as well as the clinician an unprecedented opportunity for in vivo study of the lung biology that drives normal pulmonary physiology as well as pathophysiology. PMID- 16253852 TI - 3He diffusion MRI of the lung. AB - RATIONALE AND OBJECTIVES: MR imaging of the restricted diffusion of laser polarized 3He gas provides unique insights into the changes in lung microstructure in emphysema. RESULTS: We discuss measurements of ventilation (spin density), mean diffusivity, and the anisotropy of diffusion, which yields the mean acinar airway radius. In addition, the use of spatially modulated longitudinal magnetization allows diffusion to be measured over longer distances and times, with sensitivity to collateral ventilation paths. Early results are also presented for spin density and diffusivity maps made with a perfluorinated inert gas, C3F8. METHODS: Techniques for purging and imaging excised lungs are discussed. PMID- 16253853 TI - Regional ventilation and lung mechanics using X-Ray CT. AB - Advances in computed tomographic (CT) imaging of the lung in the past decade, particularly with increased speed, resolution, gating capability, and rapidly expanding volumetric image acquisition, along with advances in image processing, have expanded the repertoire of imaging methods beyond anatomic visualization into the noninvasive study of regional lung physiological function. Recognizing that significant local disease or dysfunction can exist before global measures begin to deteriorate, the motivation for the development and application of these regional techniques is to further our understanding of the basic pathophysiological characteristics of evolving lung disease and, ultimately, develop sensitive measures for its early detection. This review emphasizes the key elements of ventilation and lung mechanics relevant for regional approaches and CT measurement principles available for their study. Examples of established and evolving methods for imaging regional ventilation and mechanics, including the xenon CT ventilation method; the relationship between changing regional CT density and air volume change; and registration-based methods for examining regional lung expansion and strain, are presented. PMID- 16253854 TI - Hyperpolarized HHe 3 MRI of the lung in cystic fibrosis: assessment at baseline and after bronchodilator and airway clearance treatment. AB - RATIONALE AND OBJECTIVES: The purpose of this study is to determine hyperpolarized helium 3 (HHe) magnetic resonance (MR) findings of the lung in patients with cystic fibrosis (CF) compared with healthy subjects and determine whether HHe MR can detect changes after bronchodilator therapy or mechanical airway mucus clearance treatment. MATERIALS AND METHODS: Thirty-one subjects, 16 healthy volunteers and 15 patients with CF, underwent HHe lung ventilation MR imaging and spirometry at baseline. Eight patients with CF then were treated with nebulized albuterol, after which a follow-up HHe MR scan was obtained. Subsequently, recombinant human deoxyribonuclease (DNase) treatment and chest physical therapy were performed in these eight subjects, followed by a third HHe MR scan. For each MR study, the number of ventilation defects was scored by a human reader. RESULTS: Patients with CF had significantly more HHe MR ventilation defects per image than healthy subjects (mean, 8.2 defects in patients with CF vs 1.6 defects in healthy subjects; P < .05). Even the four subjects with CF with a normal forced expiratory volume in 1 second had significantly more ventilation defects than healthy subjects (mean, 6.5 defects in these patients with CF; P = .0002). After treatment with albuterol, there was a small, but statistically significant, decrease in number of ventilation defects (mean, 9.6-8.0 defects; P = .025). After DNase and chest physical therapy, there was a trend toward increasing ventilation defects (mean, 8.3 defects; P = .096), but with a residual net improvement relative to baseline. CONCLUSION: In patients with CF, HHe MR ventilation defects correlate with spirometry, change with treatment, and are elevated in number in patients with CF with normal spirometry results. Thus, HHe MR appears to possess many of the characteristics required of a biomarker for pulmonary CF and may be useful in the evaluation of CF pulmonary disease severity or progression. PMID- 16253855 TI - Measurements of regional alveolar oxygen pressure using hyperpolarized 3He MRI. AB - RATIONALE AND OBJECTIVES: The aim of this work is to review hyperpolarized (HP) helium-3 (3He) magnetic resonance imaging (MRI) methods to measure regional alveolar oxygen partial pressure (P(A)O2) and oxygen depletion rate (R) in the lung. We point out limitations of the methods and suggest improvements to increase their accuracy. MATERIALS AND METHODS: P(A)O2 and R can be extracted from series of HP gas images acquired during breath hold by making use of the depolarizing effect of oxygen on HP gas. To separate oxygen-induced depolarization from other depolarizing effects, several techniques can be used. We review currently used techniques and point out their advantages and limitations. RESULTS: We show that the precision of oxygen measurements depends on a variety of parameters and can vary within the measurement volume. Accuracy of the measurement also can be influenced by diffusion of oxygen and polarized 3He and generally is different for single-slice and multislice measurements. We present numerical simulations, phantom data, and in vivo data for illustration. CONCLUSION: HP 3He MRI is a noninvasive, nonionizing, and repeatable imaging method that allows for quantitative analysis of lung function. The current techniques for measuring P(A)O2 have the potential to deliver clinically relevant functional images. PMID- 16253856 TI - Enhanced parameter estimation from noisy PET data: Part I--methodology. AB - RATIONALE AND OBJECTIVES: The reliability of positron emission tomographic (PET) images depends on the number of annihilation events that are detected. Short image durations are required to capture rapid tracer dynamics, and the resultant images are noisy. Consequently, direct parameter estimation from time-activity curves at high resolution often is unreliable. If adjacent voxels are combined into larger regions of interest the reliability of parameter estimation may be improved, but at the expense of decreased spatial resolution. In this report, a method is presented that provides an alternative to degrading image resolution. MATERIALS AND METHODS: Following the approach of Kimura et al, voxels are grouped not by spatial proximity, but by the similarity of their kinetics. Parameter estimation is performed on these groups, and derived parameters are assigned to all members of the group. Spatial information thus is preserved, but at the expense of parametric discretization. An improvement to the method of Kimura et al is described, in which data are grouped using principal components derived from artificial data. RESULTS: The application of the method is demonstrated by analysis of PET images of human lungs obtained by the nitrogen-13 infusion washout technique. In a comparison of the accuracy of parameter estimates, the enhanced method is shown to outperform the original method at all noise levels, with the difference increasing as the amount of noise increases. The robustness of this parameter estimation method in the presence of noise is described in part II of this report in this issue of Academic Radiology. CONCLUSION: A method is described that provides demonstrably robust parameter estimates from noisy PET data, while not compromising image resolution. PMID- 16253857 TI - Enhanced parameter estimation from noisy PET data: Part II--evaluation. AB - RATIONALE AND OBJECTIVES: Positron emission tomography (PET) is a minimally invasive imaging modality that provides three-dimensional distribution data for a radioactive tracer concentration within the body. Local functional parameters are estimated from these images by fitting tracer kinetic data with mathematical models. However, in some applications, the reliability of parameter estimates may be hindered by the presence of noise. In the accompanying report in this issue of Academic Radiology, a novel method using principal component analysis (PCA) was presented and used for deriving parametric images of lung function from imaged tracer kinetics of intravenously injected nitrogen 13 (13NN) in saline solution. The PCA method averages 13NN concentrations from groups of voxels (volume elements) selected for their similarity in kinetics, rather than their spatial proximity. The goal of this study is to conduct a Monte Carlo simulation to evaluate the robustness to noise of parameters derived by means of the PCA method. MATERIALS AND METHODS: This evaluation involved: (1) generating "noise free" synthetic PET images from experimental PET data, (2) adding noise to these images, (3) applying the PCA method to yield parametric images, and (4) comparing these parametric images with original noise-free images. RESULTS: Local parameters recovered by using the PCA method deviated from noise-free parameters on average by less than 1% for up to 32-fold of expected noise levels. These deviations were much less than those (>10%) recovered by using a direct curve fitting method. CONCLUSION: The novel PCA approach provides robust parametric lung functional images while preserving the spatial resolution of the original images. PMID- 16253858 TI - Quantitative computed tomography of chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) is described as airflow limitation that is not fully reversible. Quantitative assessment of structural changes within the lung that are responsible for this airflow limitation has relied on the examination of tissue obtained from surgical or postmortem specimens. However, in the past two decades, researchers have developed novel and robust tools to measure the structure of the lung parenchyma and airway wall by using computed tomographic (CT) scans, which do not require the removal of lung tissue. These techniques are extremely important because they allow longitudinal studies of the pathogenesis of COPD and the assessment of therapeutic interventions. Another application of this approach is that it potentially allows phenotyping of individuals who predominately have emphysema or small-airway disease, which may be important for the evaluation of pathogenesis and prescription of treatment options. This review describes some of these CT techniques for quantitative assessment of lung structure. PMID- 16253859 TI - Investigation of the relative effects of vascular branching structure and gravity on pulmonary arterial blood flow heterogeneity via an image-based computational model. AB - RATIONALE AND OBJECTIVES: A computational model of blood flow through the human pulmonary arterial tree has been developed to investigate the relative influence of branching structure and gravity on blood flow distribution in the human lung. MATERIALS AND METHODS: Geometric models of the largest arterial vessels and lobar boundaries were first derived using multidetector row x-ray computed tomography (MDCT) scans. Further accompanying arterial vessels were generated from the MDCT vessel endpoints into the lobar volumes using a volume-filling branching algorithm. Equations governing the conservation of mass and momentum were solved within the geometric model to calculate pressure, velocity, and vessel radius. Blood flow results in the anatomically based model, with and without gravity, and in a symmetric geometric model were compared to investigate their relative contributions to blood flow heterogeneity. RESULTS: Results showed a persistent blood flow gradient and flow heterogeneity in the absence of gravitational forces in the anatomically based model. Comparison with flow results in the symmetric model revealed that the asymmetric vascular branching structure was largely responsible for producing this heterogeneity. Analysis of average results in varying slice thicknesses illustrated a clear flow gradient because of gravity in "lower resolution" data (thicker slices), but on examination of higher resolution data, a trend was less obvious. CONCLUSIONS: Results suggest that although gravity does influence flow distribution, the influence of the tree branching structure is also a dominant factor. These results are consistent with high resolution experimental studies that have demonstrated gravity to be only a minor determinant of blood flow distribution. PMID- 16253862 TI - Why is the recent research regarding non-specific pain so non-specific? PMID- 16253863 TI - The T4 syndrome. PMID- 16253864 TI - Difficult times for Alzheimer's treatments. PMID- 16253865 TI - Weeding out new drugs. PMID- 16253866 TI - Viagra eases symptoms of pulmonary arterial hypertension in children. PMID- 16253867 TI - Unusual suspect for antipsychotic-induced diabetes. PMID- 16253868 TI - Rob Ashley of AmpliMed discusses the discovery and development of Imexon. Interview by Steve Carney. PMID- 16253869 TI - Innovation drives success in Switzerland's biotech scene. PMID- 16253870 TI - Angiogenesis inhibitors: an upcoming therapy for cancer and wet age-related macular degeneration. PMID- 16253871 TI - Broad spectrum immune monitoring in immune-mediated inflammatory disorders. PMID- 16253872 TI - The trend is innovation. PMID- 16253873 TI - A time for reflection: reviewing objectives and revising strategy. PMID- 16253874 TI - Principles and applications of LC-MS in new drug discovery. AB - The use of high-performance liquid chromatography combined with mass spectrometry (HPLC-MS) or tandem mass spectrometry (HPLC-MS-MS) has proven to be the analytical technique of choice for most assays used in various stages of new drug discovery. A summary of the key components of HPLC-MS systems, as well as an overview of major application areas that use this technique as part of the drug discovery process, will be described here. This review will also provide an introduction into the various types of mass spectrometers that can be selected for the multiple tasks that can be performed using LC-MS as the analytical tool. The strategies for optimizing the use of this technique and also the potential problems and how to avoid them will be highlighted. PMID- 16253875 TI - Automation and validation of DNA-banking systems. AB - DNA banking is one of the central capabilities on which modern genetic research rests. The DNA-banking system plays an essential role in the flow of genetic data from patients and genetics researchers to the application of genetic research in the clinic. Until relatively recently, large collections of DNA samples were not common in human genetics. Now, collections of hundreds of thousands of samples are common in academic institutions and private companies. Automation of DNA banking can dramatically increase throughput, eliminate manual errors and improve the productivity of genetics research. An increased emphasis on pharmacogenetics and personalized medicine has highlighted the need for genetics laboratories to operate within the principles of a recognized quality system such as good laboratory practice (GLP). Automated systems are suitable for such laboratories but require a level of validation that might be unfamiliar to many genetics researchers. In this article, we use the AstraZeneca automated DNA archive and reformatting system (DART) as a case study of how such a system can be successfully developed and validated within the principles of GLP. PMID- 16253876 TI - Microfluidic technologies in drug discovery. AB - Microfluidic systems are increasingly used as tools in various stages of the drug discovery process. Microscale systems offer several obvious advantages, such as low sample consumption and significantly reduced analysis or experiment time. These technologies raise the possibility of massive parallelization and concomitant reduction in cost per acquired data point. In addition, fluids in confined spaces display unique behaviors that can be used to acquire information not accessible using macroscopic systems. This article will focus on the implementation of microfluidic systems and technologies in the process of drug discovery. PMID- 16253877 TI - Biology calls the targets: combining RNAi and disease biology. AB - Target-based drug discovery starts with the identification of target genes and their respective protein products (associated with or controlling a disease relevant phenotype) that, when inhibited or activated, ameliorate the associated disease. To identify disease-relevant genes, robust tools are needed to allow biology-driven target discovery and validation. Moreover, insight into the underlying biology of a disease is essential to model a disease in vitro. Key questions are: What are the disease hallmarks? What are, from a biological point of view, the best points for therapeutic intervention? How can scientists model these points in vitro? What is the desired target profile? The closer the cellular models resemble the disease situation, the better the target profile will be. The profile is the set of biological data needed to accept the target for drug discovery. In this review, a focused approach for target discovery and validation is presented. Arrayed adenoviral siRNA libraries and disease-based cellular models are used that generate high-quality and functionally validated targets. PMID- 16253878 TI - Modelling enzyme reaction mechanisms, specificity and catalysis. AB - Modern modelling methods can now give uniquely detailed understanding of enzyme catalyzed reactions, including the analysis of mechanisms and the identification of determinants of specificity and catalytic efficiency. A new field of computational enzymology has emerged that has the potential to contribute significantly to structure-based design and to develop predictive models of drug metabolism and, for example, of the effects of genetic polymorphisms. This review outlines important techniques in this area, including quantum-chemical model studies and combined quantum-mechanics and molecular-mechanics (QM/MM) methods. Some recent applications to enzymes of pharmacological interest are also covered, showing the types of problems that can be tackled and the insight they can give. PMID- 16253880 TI - The Nobel cause. PMID- 16253881 TI - Pandemic avian influenza. PMID- 16253882 TI - Minimising antibiotic resistance. PMID- 16253883 TI - Few cheers for G8: tuberculosis in Asia. PMID- 16253884 TI - Bad breath tied to tongue. PMID- 16253886 TI - The global burden of group A streptococcal diseases. AB - The global burden of disease caused by group A streptococcus (GAS) is not known. We review recent population-based data to estimate the burden of GAS diseases and highlight deficiencies in the available data. We estimate that there are at least 517,000 deaths each year due to severe GAS diseases (eg, acute rheumatic fever, rheumatic heart disease, post-streptococcal glomerulonephritis, and invasive infections). The prevalence of severe GAS disease is at least 18.1 million cases, with 1.78 million new cases each year. The greatest burden is due to rheumatic heart disease, with a prevalence of at least 15.6 million cases, with 282,000 new cases and 233,000 deaths each year. The burden of invasive GAS diseases is unexpectedly high, with at least 663,000 new cases and 163,000 deaths each year. In addition, there are more than 111 million prevalent cases of GAS pyoderma, and over 616 million incident cases per year of GAS pharyngitis. Epidemiological data from developing countries for most diseases is poor. On a global scale, GAS is an important cause of morbidity and mortality. These data emphasise the need to reinforce current control strategies, develop new primary prevention strategies, and collect better data from developing countries. PMID- 16253887 TI - Reducing the burden of malaria in different eco-epidemiological settings with environmental management: a systematic review. AB - The public health and economic significance of malaria is enormous, and its control remains a great challenge. Many established malaria control methods are hampered by drug resistance and insecticide-resistant vectors. Malaria control measures built around environmental management are non-toxic, cost-effective, and sustainable. However, there has been no comprehensive review of the literature or meta-analysis examining the effect of these interventions. We therefore did a systematic literature review and identified 40 studies that emphasised environmental management interventions and reported clinical malaria variables as outcome measures. Of these 40 studies, environmental modification (measures aiming to create a permanent or long-lasting effect on land, water, or vegetation to reduce vector habitats--eg, the installation and maintenance of drains) was the central feature in 27 studies, environmental manipulation (methods creating temporary unfavourable conditions for the vector--eg, water or vegetation management) in four, and nine quantified the effect of modifications of human habitation. Most of the studies (n=34, 85%) were implemented before the Global Malaria Eradication Campaign (1955-69), which mainly relied on indoor residual spraying with dichlorodiphenyltrichloroethane (DDT). In 16 studies that applied environmental modification and in eight studies on modification of human habitation, the risk ratio of malaria was reduced by 88.0% (95% CI 81.7-92.1) and 79.5% (95% CI 67.4-87.2), respectively. We conclude that malaria control programmes that emphasise environmental management are highly effective in reducing morbidity and mortality. Lessons learned from these past successful programmes can inspire sound and sustainable malaria control approaches and strategies. PMID- 16253888 TI - Which delivery systems reach the poor? A review of equity of coverage of ever treated nets, never-treated nets, and immunisation to reduce child mortality in Africa. AB - Insecticide-treated nets (ITNs) and childhood vaccination are two of the most powerful interventions available to prevent childhood mortality in Africa, but ITN coverage is still very low. Current debates about how to increase ITN coverage are concerned with the roles of different supply and delivery systems, in particular whether or not commercial net markets have any useful role. Here, we review data available on coverage and equity of coverage of three interventions to prevent childhood mortality. We compiled and analysed data from nationally representative surveys in 26 African countries to compare equity of coverage of (1) the Expanded Programme on Immunisation (EPI), (2) any net, (3) ever-treated nets (ie, ITNs), and (4) never-treated nets (ie, untreated nets; UTNs). We assumed that ever-treated net coverage mostly reflects the activities of public-health programmes and projects, and that never-treated net coverage mostly reflects the activity of local unsubsidised commercial markets. We discuss the validity, limitations, and possible biases of these assumptions. We estimate that 87% of the 8.4 million children protected by nets used UTNs. We used the concentration index (CI) to assess equity of coverage of the interventions. The data shows that never-treated net coverage is surprisingly equitable: overall, and despite substantial regional variations, it is comparable in equity to EPI (median CI(UTN)=0.166, CI(EPI)=0.075; p=0.3). In almost all countries, coverage of ITNs is strongly concentrated in the least poor households, and significantly more inequitable than both UTNs (median CI(ITN)=0.435, mean CI(UTN)=0.158; p<0.001) and EPI (median CI(ITN)=0.435, CI(EPI)=0.075; p<0.001). These results suggest that the public-health value of commercial net markets has been greatly underestimated, and that these markets have so far contributed more to equitable and sustainable coverage of mosquito nets, and hence to the prevention of malaria in Africa, than have the ITNs delivered by public-health systems and projects. PMID- 16253889 TI - Understanding the symptoms of the common cold and influenza. AB - The common cold and influenza (flu) are the most common syndromes of infection in human beings. These diseases are diagnosed on symptomatology, and treatments are mainly symptomatic, yet our understanding of the mechanisms that generate the familiar symptoms is poor compared with the amount of knowledge available on the molecular biology of the viruses involved. New knowledge of the effects of cytokines in human beings now helps to explain some of the symptoms of colds and flu that were previously in the realm of folklore rather than medicine-eg, fever, anorexia, malaise, chilliness, headache, and muscle aches and pains. The mechanisms of symptoms of sore throat, rhinorrhoea, sneezing, nasal congestion, cough, watery eyes, and sinus pain are discussed, since these mechanisms are not dealt with in any detail in standard medical textbooks. PMID- 16253891 TI - Hydatid cyst of the spine. PMID- 16253890 TI - Targeting the glycans of gp120: a novel approach aimed at the Achilles heel of HIV. AB - The development of drug resistance in HIV compromises the long-term efficacy of current therapies. Furthermore, vaccine development faces huge problems, mainly because of the low antigenicity and immunogenicity of the HIV envelope glycoprotein gp120 and the efficient hiding of highly immunogenic epitopes by its glycans. There is evidence that mutant HIV strains containing glycosylation site deletions trigger the production of specific neutralising antibodies to previously hidden gp120 epitopes. I present a hypothesis that development of resistance against drugs that target the glycans on gp120 would result in a marked enhancement of neutralisation of HIV by the immune system--ie, drugs directed against the carbohydrate component of gp120 will select for mutant virus strains that progressively gain deletions in the glycosylation sites of gp120. Previously hidden epitopes would then be uncovered, and the virus will become highly susceptible to markedly increased immunological neutralisation. I believe this novel approach may become an entirely new therapeutic concept that exploits the high mutation rate of HIV and allows drug therapy to act in concert with a triggered immune response to suppress HIV more efficiently. Moreover, this approach could be applied to treat other chronic infections by viruses that contain a glycosylated envelope (eg, hepatitis B and C). PMID- 16253892 TI - Endocrine tumours of the stomach. AB - Gastric endocrine tumours (gastric carcinoids) usually grow from enterochromaffin like (ECL) cells. Three types of tumour may be distinguished on the basis of the background gastric pathology: type I, which develops in atrophic body gastritis (ABG); type II, which is associated with multiple endocrine neoplasia and Zollinger-Ellison syndrome; and the sporadic type III, which is not associated with any background pathology. This classification plays a major role in determining the optimal approach to these diseases. In fact, type I carcinoids can be considered to be benign lesions, with exceptional risk of metastases. Type II, in contrast, may be associated with distant metastases, which are also common in type III carcinoids. The therapeutic approach is based mainly on endoscopic excision and somatostatin analogues in types I and II, or on surgical resection in type III. Both types I and II grow under the stimulus of hypergastrinaemia through a well-described sequence. However, gastrin is sufficient to cause ECL cell hyperplasia and dysplasia, but not transformation, which is due to menin defects in MEN-I patients, or to other unknown alterations in ABG. Several other candidates--including Bcl2, p53 and MMP9--have been linked with carcinoid initiation and progression. The biology of type III tumours which are not associated with hypergastrinaemia is still poorly understood. PMID- 16253893 TI - Duodenal neuroendocrine tumors: Classification, functional syndromes, diagnosis and medical treatment. AB - Duodenal neuroendocrine tumors (NETs) comprise 2-3% of all GI endocrine tumors and are increasing in frequency. These include gastrinomas, somatostatinomas, nonfunctional NETs, gangliocytic paragangliomas, and poorly differentiated NE carcinomas. Although, the majority are nonfunctional, these tumors are a frequent cause of Zollinger-Ellison syndrome and can cause other clinical hormonal syndromes (carcinoid, Cushing's, etc.). In this chapter, their epidemiology, clinical aspects, localization, diagnosis and medical treatment are reviewed including the latest advances in each area. PMID- 16253894 TI - Surgery and prognosis of duodenal gastrinoma as a duodenal neuroendocrine tumor. AB - It has become increasingly clear that duodenal gastrinomas are the most common cause of Zollinger-Ellison syndrome (ZES). However, attempts to find these tumors before and during surgery for ZES have had limited success until duodenotomy (opening the duodenum) was described. The routine use of duodenotomy in patients with non-familial gastrinoma increases the number of duodenal tumors found, and the immediate and long-term cure-rate. The increase in cure-rate appears to be secondary to increased detection of small, previously undetectable duodenal gastrinomas. Duodenotomy detects small tumors (<1 cm) in the proximal duodenum. It does not detect more duodenal gastrinomas per patient, nor does it detect tumors in unusual duodenal locations. Duodenotomy decreases the death-rate associated with these tumors. However, it has not affected the rate of development of liver metastases. Duodenotomy is a critical method to find duodenal gastrinomas. It should be routinely performed in all surgery to find and remove gastrinoma for cure of ZES. PMID- 16253895 TI - Tumours of the midgut (jejunum, ileum and ascending colon, including carcinoid syndrome). AB - (Neuro-)endocrine tumours of the gastrointestinal tract are also called 'carcinoids'. (Neuro-)endocrine midgut tumours can be categorized according to their clinical behaviour. Most tumours are non-functioning. Functioning tumours are responsible for the carcinoid syndrome. The carcinoid syndrome is almost uniquely associated with midgut carcinoids. Symptoms of the carcinoid syndrome are caused by an excess of biogenic amines, peptides and other factors in the circulation. The typical symptoms of the carcinoid syndrome are diarrhoea, flushing, and carcinoid heart disease. Carcinoid heart disease involves the tricuspid and pulmonary valves and the endocardium. Serum chromogranin A and urinary excretion of 5-hydroxy-indoleacetic acid (5-HIAA) are biochemical markers. Carcinoid tumours express large numbers of high-affinity somatostatin receptors. These can bind the currently available octapeptide somatostatin analogues. In inoperable patients, biotherapy with somatostatin analogues and interferon-alpha is the treatment of choice. Somatostatin analogues and interferon-alpha significantly improve symptoms. PMID- 16253896 TI - Midgut carcinoid tumours: surgical treatment and prognosis. AB - Midgut carcinoids originating in the small intestine are the most common cause of the carcinoid syndrome. These tumours typically progress slowly and have an extended disease course, and although they often present with metastases at diagnosis, surgical treatment has become increasingly important for their management. Surgery should include efforts to remove mesenteric metastases, which may cause severe long-term abdominal complications with typical fibrotic intestinal entrapment and small-bowel ischaemia due to compression of mesenteric vessels. Attempts should also be made to surgically remove or ablate liver metastases, since this may provide considerable palliation of the carcinoid syndrome. For patients with the carcinoid syndrome surgery is combined with continuous biotherapy with long-acting somatostatin analogues, which may alleviate symptoms and stabilize disease or slow progression. Favourable survival and appreciable quality of life can be expected with this combined treatment, even in patients with advanced midgut carcinoids. PMID- 16253897 TI - Neuroendocrine tumours (carcinoids) of the appendix. AB - Neuroendocrine tumours (NETs) of the appendix (formerly 'carcinoids') are rare and are usually detected incidentally after appendectomy. Histopathologically they derive from a subepithelial cell population, which is different from NETs in other sites. They are preferentially located at the tip of the appendix. Tumours <1 cm hardly ever metastasize and are treated by appendectomy. Tumours >2 cm require right hemicolectomy because of a significant risk of metastatic spread. Treatment for lesions 1-2 cm is controversial and needs further characterization of the tumour (i.e. mesoappendiceal invasion, vascular invasion, mitotic activity, proliferation markers) and careful patient risk evaluation. Goblet-cell carcinoids have features resembling both carcinoid and adenocarcinoma and should be treated by hemicolectomy. Overall prognosis of small appendiceal NET is excellent in all ages. PMID- 16253898 TI - Endocrine tumours of the hindgut. AB - Neuroendocrine tumours of the colon and rectum are rare but distinct with regard to clinical symptoms, diagnostic and therapeutic management and prognosis compared to other neuroendocrine tumours of the gut as well as ordinary colorectal cancer. Therapeutic algorithms are proposed depending mainly on analogous TNM categories and grading considering conventional and experimental surgical and non-surgical therapy. Colonic neuroendocrine tumours are often misdiagnosed as undifferentiated adenocarcinoma and are therefore not properly treated with adjuvant and additive chemotherapy. As most rectal neuroendocrine tumours are benign because of submucosal extension only, the size and infiltration depth correlates with lymph-node and distant metastases and therefore with the prognosis. It is unknown whether endoscopic ultrasound can improve the diagnostic accuracy compared to size-related conclusions, and therefore whether it can change therapeutic strategies and improve survival by modern rectal surgery. PMID- 16253899 TI - Endocrine tumours of the pancreas. AB - Endocrine pancreatic tumours (EPTs) are uncommon tumours occurring in approximately 1 in 100,000 of the population, representing 1-2% of all pancreatic neoplasms. Some of the tumours may be part of multiple endocrine neoplasia type one (MEN-1) syndrome or von Hippel-Lindau (vHL) disease. EPTs are classified as functioning or non-functioning tumours on the basis of their clinical manifestation. The biochemical diagnosis of EPT is based on hormones and amines released. Besides specific markers such as insulin, there are also general tumour markers such as chromogranin A, which is the most valuable marker and has been reported to be increased in plasma in 50-80% of patients with EPTs and correlates with tumour burden. The location of endocrine tumours of the pancreas includes different techniques, from endoscopic investigations to scintigraphy (e.g. somatostatin receptor scintigraphy) and positron emission tomography. The medical treatment of endocrine pancreatic tumours consists of chemotherapy, somatostatin analogues and alpha-interferon. None of these can cure a patient with malignant disease. In future, therapy will be custom-made and based on current knowledge of tumour biology and molecular genetics. PMID- 16253900 TI - Insulinoma. AB - Although rare, insulinomas are the most common functioning islet cell tumour of the pancreas. Recognition of the key neuroglycopenic symptoms should trigger the initial investigation. Biochemical proof of endogenous hyperinsulinemic hypoglycemia establishes the diagnosis. Several options are available for imaging and localizing these tumours including ultrasonography, computed tomography, and intra-arterial calcium stimulation with venous sampling. The tumours are usually small, single, benign, well-circumscribed, and evenly distributed throughout the pancreas. This tumour may be a part of the multiple endocrine neoplasia type 1 (MEN-1) syndrome, in which case the tumours are almost always multiple. Surgical treatment is the only curative method, traditionally accomplished with enucleation or partial pancreatic resection. Patients are almost invariably cured lifelong with complete excision of a benign insulinoma. The most recent developments in this area are the recognition of noninsulinoma pancreatogenous hypoglycemia syndrome as a cause of organic hypoglycemia, and the development of laparoscopic techniques to excise these tumours. PMID- 16253901 TI - Surgical treatment and prognosis of gastrinoma. AB - Zollinger-Ellison syndrome (ZES) is a clinical syndrome with severe peptic ulcer disease and diarrhea caused by gastric acid hypersecretion secondary to a neuroendocrine tumour that secretes excessive amounts of the hormone gastrin (gastrinoma). Gastrinomas occur in a familial and a sporadic form. Patients with gastrinoma in the familial setting of Multiple Endocrine Neoplasia type 1 (MEN-1) are seldom, if ever, cured of Zollinger-Ellison syndrome by the current non Whipple operations to remove duodenal and pancreatic gastrinoma. Surgery is currently used in these patients to deal with the malignant nature of pancreatic or duodenal neuroendocrine tumours. Malignant potential is best determined by tumour size. Tumours that are greater than 2 cm in size should be excised. In the sporadic setting, cure occurs in a significant proportion of patients (50%) by surgical resection of gastrinoma. Duodenotomy has improved both the tumour detection rate and the cure rate and should be routinely done. Whipple pancreaticoduodenectomy results in the highest probability of cure in both sporadic and MEN-1 gastrinoma patients as it removes the entire gastrinoma triangle. However, the excellent long-term survival of these patients with lesser operations and the increased operative mortality and long-term morbidity of Whipple make its current role unclear until further studies are done. PMID- 16253902 TI - Rare endocrine tumours of the GI tract. AB - Functional pancreatic endocrine tumors other than gastrinoma and insulinoma are quite rare. The principles of management include the diagnosis and management of the functional hormonal syndrome, and management of the potentially malignant tumor. Optimally, control of the hormonal syndrome is achieved preoperatively to stabilize the patient status for the operation, however, resection may be an important part of the control of the hormonal syndrome. Ultimately, the only curative treatment for these neoplasms is complete tumour resection, when feasible. PMID- 16253903 TI - Pancreatic tumours as part of the MEN-1 syndrome. AB - Pancreaticoduodenal tumours (PETs) occur in a majority of MEN-1 patients, and have appeared as a major cause of disease-related death. Previous discussions about treatment have mainly dealt with management of various functioning tumours and clinical syndromes of hormone excess. However, hormonal syndromes often occur late with MEN-1 pancreatic tumours, and when developed indicate presence of metastases in up to 50% of the patients. Prospective screening is therefore recommended in MEN-1 with biochemical markers and endoscopic ultrasound for early detection of PETs, and early surgery before metastases have developed. Surgery is recommended in patients with or without hormonal syndromes in absence of disseminated liver metastases. The suggested operation includes enucleation of tumours in the head of the pancreas, excision of duodenal gastrinomas together with clearance of lymph node metastases, and distal 80% subtotal pancreatic resection as prophylaxis against tumour recurrence. This strategy with early and aggressive surgery is believed to reduce the risks for malignant progression. PMID- 16253905 TI - The impact of respiration on left atrial and pulmonary venous anatomy: implications for image-guided intervention. AB - BACKGROUND: Image-guided intervention using pre-acquired CT/MR 3-dimensional images is an emerging strategy for atrial fibrillation (AF) ablation but may be limited by its use of static images to depict dynamic physiology. The effect of biologic factors such as respiration on the left atrial-pulmonary venous (LA-PV) anatomy is not well understood but is likely to have important implications. Conventional CT/MR imaging is performed during an inspiratory breath-hold, while electroanatomical mapping (EAM) during "quiet" breathing approximates an expiratory breath-hold. This study examined the effects of respiration on LA-PV anatomy and the error introduced by respiration on the integration of EAM with 3D MR imaging. METHODS: Pre-procedural MRI angiography was performed at both end expiration (EXP) and end-inspiration (INSP) in 20 patients undergoing AF catheter ablation. 3D INSP and EXP surface reconstructions of the LA-PVs were compared. In selected pts, EAM data acquired during the ablation procedure (n=7) were integrated with the 3D MRI datasets. RESULTS: Qualitative assessment of the INSP and EXP 3D images revealed splaying of the PVs and reduction in PV caliber of the right-sided PVs during held inspiration. After aligning these two datasets, the average surface-to-surface distance calculated by region ranged from 1.99mm (right middle PV) to 3.79mm (left superior PV). Registration of the EAM to the MRI models was better for the EXP dataset (2.30+/-0.73mm) than the INSP dataset (3.03+/-0.57mm; p=0.004). CONCLUSION: There are significant changes in LA-PV anatomy with respiration. MR images acquired during standard held inspiration may introduce unnecessary errors in registration during image-guided intervention. PMID- 16253904 TI - Randomized comparison of encircling and nonencircling left atrial ablation for chronic atrial fibrillation. AB - BACKGROUND: Left atrial (LA) circumferential ablation has been reported to eliminate atrial fibrillation (AF). Whether an ablation without encirclement of the pulmonary veins (PVs) is as effective as LA circumferential ablation is not clear. OBJECTIVES: The purpose of this study was to compare the efficacy of LA circumferential ablation and nonencircling linear ablation in patients with chronic AF. METHODS: Eighty patients with chronic AF were randomized to undergo LA circumferential ablation (n = 40) or nonencircling linear ablation (n = 40). In LA circumferential ablation, the PVs were encircled, with additional lines made in the mitral isthmus and posterior wall or roof. In nonencircling linear ablation, 4 +/- 1 ablation lines were created through areas of complex electrograms, with lines in the roof (38), anterior wall (36), septum (40), mitral isthmus (32), and posterior annulus (6). The endpoint of LA circumferential ablation and nonencircling linear ablation was voltage abatement. RESULTS: LA flutter occurred in 15% after LA circumferential ablation and in 18% after nonencircling linear ablation (P = .8). A repeat ablation procedure was performed for recurrent AF in 7 and 11 patients or for atrial flutter in 6 and 4 patients after LA circumferential ablation and nonencircling linear ablation, respectively (P = .8). At 9 +/- 4 months, the prevalence of AF was 28% in the LA circumferential ablation and 25% in the nonencircling linear ablation group (P = .8). Sixty-eight percent and 60% of patients were in sinus rhythm and free of AF and atrial flutter in the absence of antiarrhythmic drug therapy after LA circumferential ablation and nonencircling linear ablation, respectively (P = .5). There were no complications. CONCLUSION: Nonencircling linear ablation and LA circumferential ablation are equally efficacious in eliminating chronic AF. However, the advantage of nonencircling linear ablation is that it eliminates the need for ablation along the posterior wall of the LA. Therefore, nonencircling linear ablation may avoid the small but real risk of atrioesophageal fistula formation associated with LA circumferential ablation. PMID- 16253906 TI - Prolongation of the fast pathway effective refractory period during cryoablation in children: a marker of slow pathway modification. AB - BACKGROUND: The fast pathway effective refractory period (ERP) has been reported to decrease after slow pathway modification with radiofrequency (RF) energy. How the fast pathway ERP changes during the ablation application has not been reported with either RF or cryoenergy. OBJECTIVES: Using the unique features of cryotherapy, this study assesses the short-term changes in fast pathway ERP during cryomodification of the slow pathway and examines whether these changes are a useful marker for successful slow pathway modification in children. METHODS: Nineteen pediatric patients (median age 15.1 years, range 9.6-19.6 years; weight 60.7 kg, range 35.6-130.2 kg) with anterograde dual AV nodal physiology underwent slow pathway modification with catheter-based cryoablation. Programmed stimulation was performed during cryoapplications after reaching -25 degrees C to assess fast pathway and slow pathway conduction. Data were analyzed from 59 of 237 cryoapplications where the fast pathway ERP was measured more than once (n = 13 patients). RESULTS: For 23 of 59 applications where the slow pathway was modified, the fast pathway ERP significantly increased during cryotherapy (Delta = 33.5 ms, P <.0001). The magnitude of fast pathway ERP prolongation during cryotherapy was larger when the slow pathway was modified than when there was no effect on slow pathway conduction (33.5 +/- 30.5 vs 5.8 +/- 18.9 ms, P =.0005). Prolongation of fast pathway ERP by >/=20 ms had 70% sensitivity and 72% specificity for predicting slow pathway modification. Following termination of cryoapplications, which resulted in slow pathway modification, the fast pathway ERP had significantly decreased from baseline (difference 44.5 ms, P <.0001). The effect on fast pathway ERP was not related to changes in cycle length during (R(2) = 0.04, P = .045) or after ablation (R(2) = 0.13, P = .012). CONCLUSION: The fast pathway ERP prolongs during cryoapplications that result in slow pathway modification and shortens after termination of cryoapplications. The magnitude of fast pathway ERP prolongation during cryoapplication may be useful as a marker for successful slow pathway modification. PMID- 16253907 TI - Modification of the slow pathway in atrioventricular nodal reentrant tachycardia: no pain, no gain. PMID- 16253908 TI - Predictors of appropriate implantable defibrillator therapies in patients with arrhythmogenic right ventricular dysplasia. AB - BACKGROUND: Arrhythmogenic right ventricular dysplasia (ARVD) is an inherited cardiomyopathy characterized by ventricular arrhythmias and sudden cardiac death. The risk factors for sudden death and indications for implantable cardioverter defibrillator (ICD) placement in patients with ARVD are not well defined. OBJECTIVES: The purpose of this study was to determine which clinical and electrophysiologic variables best predict appropriate ICD therapies in patients with ARVD. Particular attention focused on whether the ICD was implanted for primary or second prevention. METHODS: We enrolled 67 patients (mean age 36 +/- 14 years) with definite or probable ARVD who had undergone ICD placement. Appropriate ICD therapies were recorded, and Kaplan-Meier analysis was used to compare the event-free survival time between patients based upon the indication for ICD placement (primary vs secondary prevention), results of electrophysiologic testing, and whether the patient had probable or definite ARVD. RESULTS: Over a mean follow-up of 4.4 +/- 2.9 years, 40 (73%) of 55 patients who met task force criteria for ARVD and 4 (33%) of 12 patients with probable ARVD had appropriate ICD therapies for ventricular tachycardia/ventricular fibrillation (VT/VF; P = .027). Mean time to ICD therapy was 1.1 +/- 1.4 years. Eleven of 28 patients who received an ICD for primary prevention (39%) and 33 of 35 patients who received an ICD for secondary prevention (85%) experienced appropriate ICD therapies (P = .001). Electrophysiologic testing did not predict appropriate ICD interventions in patients who received an ICD for primary prevention. Fourteen patients (21%) received ICD therapy for life-threatening (VT/VF >240 bpm) arrhythmias. There was no difference in the incidence of life-threatening arrhythmias in the primary and secondary prevention groups (P = .29). CONCLUSION: Patients who meet task force criteria for ARVD are at high risk for sudden cardiac death and should undergo ICD placement for primary and secondary prevention, regardless of electrophysiologic testing results. Further research is needed to confirm that a low-risk subset of patients who may not require ICD placement can be identified. PMID- 16253909 TI - Reentrant and nonreentrant focal left atrial tachycardias occur after pulmonary vein isolation. AB - BACKGROUND: Both focal and macroreentrant atrial tachycardia (ATs) can occur after pulmonary vein (PV) isolation for treatment of atrial fibrillation (AF). We report the response to pacing and pharmacologic maneuvers performed in patients with stable focal ATs after segmental PV isolation. OBJECTIVES: The purpose of this study was to determine the mechanism of focal ATs occurring after PV isolation. METHODS: Patients with persistent left AT after cessation of antiarrhythmic drug therapy presented for mapping and ablation. Electroanatomic mapping during AT was performed. Entrainment was performed from multiple right and left atrial sites. Single-beat resetting was performed. Adenosine was infused intravenously to determine the effect on the tachycardia. RESULTS: Five patients (3 men and 2 women; age 65 +/- 10 years) had focal left AT that persisted in response to pacing maneuvers. Four patients had ATs (cycle length 265 +/- 18 ms) that demonstrated focal areas of early activation at the septal aspect of the right lower (3 patients) or right upper (1 patient) PV ostium. Resetting demonstrated a flat-plus increasing curve in two patients. Adenosine was infused with transient AV block and no change to the tachycardia cycle length in three patients. Tachycardias were entrained from multiple left atrial sites. Recordings from the ablation catheter at the critical isthmus typically demonstrated mid diastolic or long fractionated potentials. One tachycardia with a longer and more variable cycle length (480-598 ms) did not demonstrate fusion during pacing from distant sites. CONCLUSION: Persistent focal left ATs may occur after segmental PV isolation. Many of these tachycardias are caused by a focal reentrant circuit located at the PV ostium; however, focal nonreentrant rhythms also may occur. PMID- 16253910 TI - Automatic measurement of atrial pacing thresholds in dual-chamber pacemakers: clinical experience with atrial capture management. AB - BACKGROUND: The Medtronic EnPulse pacemaker incorporates the new atrial capture management (ACM) algorithm to automatically measure atrial capture thresholds and subsequently manage atrial pacing outputs. OBJECTIVES: The purpose of this study was to evaluate the clinical performance of ACM. METHODS: Two hundred patients with an indication for a dual-chamber pacemaker underwent implantation. ACM thresholds and manually measured atrial pacing thresholds were assessed at follow up visits. Clinical equivalence was defined as the ACM-measured threshold being within -0.25 V to +0.5 V of the manually measured threshold. The clinician analyzed all ACM measurements performed in-office for evidence of proarrhythmia. RESULTS: All 200 implanted patients had a 1-month visit, and validated manual and in-office ACM threshold data were available for 123 patients. The ACM threshold was 0.595 +/- 0.252 V, and the manual threshold was 0.584 +/- 0.233 V. The mean difference was 0.010 V with a 95% confidence interval of (-0.001, 0.021). The mean difference over all visits was 0.011 V. For all patients, the individual threshold differences were within the range of clinical equivalence at all visits. No atrial arrhythmias were observed during 892 ACM tests in 193 patients. CONCLUSION: This study demonstrated that the ACM algorithm is safe, accurate, and reliable over time. ACM was demonstrated to be clinically equivalent to the manual atrial threshold test in all patients at 1 month and over the entire follow-up period of up to 6 months. ACM ensures atrial capture, may save time during follow-up, and can be used to manage atrial pacing outputs. PMID- 16253911 TI - Extent of myocardial viability predicts response to biventricular pacing in ischemic cardiomyopathy. AB - BACKGROUND: The clinical response to biventricular pacing is unpredictable, especially in patients with ischemic cardiomyopathy. OBJECTIVES: The purpose of this study was to prospectively examine the relationship between the extent of myocardial viability and the response to cardiac resynchronization therapy. METHODS: Twenty-one patients with ischemic left ventricular (LV) dysfunction (left ventricular ejection fraction [LVEF] 21 +/- 5%), New York Heart Association (NYHA) functional class III-IV, and QRS >120 ms received biventricular devices. Myocardial viability was assessed by myocardial contrast echocardiography, and a perfusion score index (PSI) was calculated from summed segmental perfusion scores. LV performance was assessed by echocardiography on the day after implantation and at 6 months. RESULTS: PSI was closely correlated with acute improvement in LVEF (P = .003, r = 0.65), stroke volume (P = .02, r = 0.54), and end-systolic volume (P = .05, r = -0.49). PSI also correlated with early diastolic LV relaxation (E', P < .05, r = 0.50) and global myocardial performance or Tei index (P = .003, r = 0.63). By multiple linear regression analysis, PSI provided incremental predictive value to the degree of dyssynchrony, measured by tissue Doppler imaging, for predicting improvement in LVEF. At 6 months, PSI remained positively correlated with improvement in ventricular performance and with reduction in LV end-diastolic dimension (P = .003, r = -0.68). PSI also influenced the clinical variables of NYHA class, 6-minute walk distance, quality of-life score, and number of hospitalizations for heart failure. CONCLUSION: In patients with ischemic cardiomyopathy, the extent of myocardial viability predicts acute and long-term improvement in LV performance, exercise tolerance, and reduction in LV end-diastolic dimension with biventricular pacing. PMID- 16253912 TI - Targeted mutational analysis of ankyrin-B in 541 consecutive, unrelated patients referred for long QT syndrome genetic testing and 200 healthy subjects. AB - BACKGROUND: Mutations in ANK2-encoded ankyrin-B underlie long QT syndrome type 4 (LQT4) and various other dysrhythmia phenotypes. OBJECTIVES: The purpose of this study was to determine the prevalence and spectrum of ankyrin-B mutations in a large cohort of unrelated patients referred for LQTS genetic testing and among healthy control subjects. METHODS: Between August 1997 and July 2004, 541 consecutive, unrelated patients (358 females, average age at diagnosis 24 years, average QTc 482 ms) were referred to Mayo Clinic's Sudden Death Genomics Laboratory for comprehensive mutational analysis of the five cardiac channel genes implicated in LQTS: KCNQ1 (LQT1), KCNH2 (LQT2), SCN5A (LQT3), KCNE1 (LQT5), and KCNE2 (LQT6). Based on this prior analysis, 269 of 541 cases lacked an identifiable mutation (genotype negative). In this study, targeted mutational analysis of 10 ANK2 exons (36,37,39-46) encoding the critical C-terminal regulatory domain or implicated previously as hosting pathogenic mutations was performed on genomic DNA from 541 patients and 200 control subjects using polymerase chain reaction, denaturing high-performance liquid chromatography, and direct DNA sequencing. RESULTS: Overall, 14 distinct nonsynonymous variants (10 novel) were observed in 9 (3.3%) of 269 genotype-negative LQTS patients, 5 (1.8%) of 272 genotype-positive LQTS cases, 4 (4%) of 100 white controls, and 9 (9%) of 100 black controls. Four variants found in controls (L1622I, T1626N, R1788W, and E1813K) were implicated previously as LQT4-associated mutations and displayed functional perturbations in vitro. All genotype-negative LQTS cases hosting ANK2 variants had been diagnosed as "atypical" or "borderline" cases, most presenting with normal QTc, nonexertional syncope, U waves, and/or sinus bradycardia. CONCLUSION: Nonsynonymous ankyrin-B variants were detected in nearly 3% of unrelated LQTS patients and nearly 7% of healthy control subjects. Genotype negative LQTS patients with a single ANK2 variant displayed nonexertional syncope, U waves, sinus bradycardia, and extracardiac findings. Whether the identification of previously reported functionally significant variants residing in 2% of apparently healthy subjects suggests proarrhythmic potential or potential misclassification warrants further scrutiny. PMID- 16253913 TI - Decrease in fluoroscopic cardiac silhouette excursion precedes hemodynamic compromise in intraprocedural tamponade. AB - BACKGROUND: Acute pericardial tamponade is a life-threatening complication of invasive cardiovascular procedures. Survival depends on early recognition and treatment. A diagnostic test to detect pericardial fluid accumulation before a significant fall in blood pressure and without contamination of the sterile field would be valuable. OBJECTIVE: We tested the hypothesis that fluoroscopic excursion of the cardiac silhouette decreases early in the course of acute pericardial tamponade and precedes hemodynamic compromise. METHODS: The pericardial space of seven pigs was accessed by a sub-xiphoid puncture. Tamponade was produced by intrapericardial saline infusion at 20-25 mL/minute until the pericardial pressure equalized with right ventricular end diastolic pressure or the systolic blood pressure reached 40 mmHg. Supine fluoroscopic images were obtained every 2 minutes in the left anterior oblique view with simultaneous echocardiography. The fluoroscopic heart silhouette was digitized, and the maximum excursion during the cardiac cycle was quantified by custom software. The qualitative excursion of the fluoroscopic heart silhouette on randomly selected video images was also graded by two independent observers who were blinded to the time course of the experiment and the hemodynamics. RESULTS: During progressive pericardial fluid accumulation, the cardiac silhouette excursion quantified by the custom software (p < 0.001) and by video rating (p < 0.0001) was significantly reduced within 2 minutes. A statistically significant fall in blood pressure compared with baseline did not occur until 6 minutes (89 +/- 21 vs. 121 +/- 15 mmHg, p < 0.001). The interobserver agreement was very close, with a kappa statistic of 0.78. The reduction in cardiac silhouette excursion was apparent as soon as the effusion was detected by echocardiography. CONCLUSION: Cardiac silhouette excursion becomes reduced early in the course of acute pericardial fluid accumulation. This fluoroscopic observation can be used to detect impending pericardial tamponade before hemodynamic collapse. PMID- 16253914 TI - Atrioventricular nodal reverse facilitation in connexin40-deficient mice. AB - BACKGROUND: Facilitation is an important physiologic property of the atrioventricular (AV) node. Previous studies demonstrated abnormal AV conduction in connexin (Cx)40-deficient mice. OBJECTIVES: We hypothesize that Cx40-deficient mice display altered patterns of AV nodal facilitation compared with wild-type mice. METHODS: Sixteen 36-week-old mice (eight Cx40(-/-) mice and eight Cx40(+/+) controls) underwent in vivo closed chest electrophysiologic study. A 2Fr octapolar catheter was advanced into the right ventricle to record a His-bundle electrogram. A special facilitation stimulation protocol was performed in each mouse to evaluate facilitation. Following atrial drive pacing (S1S1) at 150 ms, a facilitating beat S2 was delivered prior to the test beat S3. S3H3 was measured for varying S1S2 values at fixed H2S3 intervals. RESULTS: Progressive shortening of S1S2 (from 150 ms to 130, 110, and 90 ms) resulted in gradual prolongation of S2H2. The prolongation was more pronounced in Cx40(-/-) mice for each S1S2 compared with wild-type mice (P <.001). In each wild-type mouse, for a given H2S3 interval, this gradual increase in S2H2 produced progressive shortening of S3H3, so-called AV nodal facilitation phenomenon. However, in each Cx40(-/-) mouse, facilitation was seen only at S1S2 of 130 ms (P <.001 vs S1S2 of 150 ms). Evidence of reverse facilitation was documented at S1S2 of 110 and 90 ms. CONCLUSION: Facilitation is observed in wild-type mice. With similar S1S2 intervals in Cx40-deficient mice, facilitation is seen only at longer S1S2 intervals, whereas reverse facilitation is seen at shorter S1S2 intervals, suggesting that Cx40 is involved in the generation of AV nodal facilitation. PMID- 16253915 TI - Functional assessment of compound mutations in the KCNQ1 and KCNH2 genes associated with long QT syndrome. AB - BACKGROUND: Long QT syndrome (LQTS) is a cardiovascular disorder characterized by prolonged QTc time, syncope, or sudden death caused by torsades de pointes and ventricular fibrillation. We investigated the clinical and electrophysiologic phenotype of individual mutations and the compound mutations in a family in which different genotypes could be found. OBJECTIVES: The purpose of this study was to determine the impact of genotype-based diagnostic assessment in LQTS. METHODS: We used cascade screening and functional analyses to investigate the phenotype in a family with LQTS. The contributions of the compound mutations in the KCNQ1 and KCNH2 genes (KCNQ1 R591H, KCNH2 R328C) were analyzed by heterologous expression in Xenopus laevis oocytes using two-electrode voltage clamp and by confocal imaging. RESULTS: KCNH2 R328C did not show any functional phenotype whereas KCNQ1 R591H resulted in severe reduction of current. Neither wild-type nor mutant channels affected each other functionally in coexpression experiments. Therefore, a direct interaction between KCNQ1 and KCNH2 was ruled out under these conditions. CONCLUSION: Assessment of novel mutational findings in LQTS should include accurate genetic and functional analysis. Notably, appropriate studies are needed if two or more mutations in different genes are present in one proband. Our findings prompt reconsideration of the impact of compound mutations in LQTS families and reinforce the need for thorough functional evaluation of novel ion channel mutations before assignment of pathogenic status. PMID- 16253916 TI - Electrocardiographic imaging (ECGI), a novel diagnostic modality used for mapping of focal left ventricular tachycardia in a young athlete. AB - We report the first clinical application of electrocardiographic imaging (ECGI), a new, noninvasive imaging modality for arrhythmias, in an athlete with focal ventricular tachycardia (VT) originating from a left ventricular (LV) diverticulum. A reconstructed map of the epicardial activation sequence during a single premature ventricular complex (PVC) of an identical QRS morphology to the clinical VT, generated from 224-electrode body surface ECGs and a chest CT (ECGI), localized the PVC to the site of the diverticulum. This correlated with subsequent maps obtained using standard techniques. We describe the first case that used ECGI to guide diagnosis and therapy of a clinical tachyarrhythmia. PMID- 16253917 TI - Inappropriate therapy due to triple counting of the ventricular electrogram in a patient with implantable cardioverter-defibrillator. PMID- 16253918 TI - Spontaneous migration of the site of 2:1 atrioventricular block during atrioventricular nodal reentrant tachycardia. PMID- 16253919 TI - What can nonlinear dynamics teach us about the development of ventricular tachycardia/ventricular fibrillation? PMID- 16253920 TI - Irregular, nonsustained paroxysmal supraventricular tachycardia: what is the mechanism? PMID- 16253921 TI - Delivery of antitachycardia pacing after a full-energy shock during the same ventricular tachycardia episode: appropriate device function? PMID- 16253922 TI - How to analyze T-wave alternans. PMID- 16253923 TI - Intracardiac echocardiographic imaging of the left atrial appendage. PMID- 16253924 TI - Direct visualization of the coronary sinus ostium and branches with the flexible endoscope. PMID- 16253926 TI - Drug-induced proarrhythmia and use of QTc-prolonging agents: clues for clinicians. AB - Use of drugs with the potential for prolongation of the QTc interval and proarrhythmia is a growing challenge facing clinicians. Many pharmaceutical agents have been denied approval for human use, approved with restrictions and warnings regarding proarrhythmia, or withdrawn from the market based upon arrhythmic risk. Despite known risk factors for QTc prolongation and drug-induced arrhythmia, precise prediction of the risk of torsades de pointes (TdP) in an individual patient remains difficult. The mechanism of drug-induced TdP typically involves use of an agent that blocks the I(Kr) cardiac potassium current, often in combination with risk-amplifying factors such as high drug levels, reduced drug metabolism, polypharmacy, and patient-specific factors such as gender, age, and genetic polymorphism. For the clinician, an integrated approach involving appreciation of the risk factors for proarrhythmia combined with computer-based risk assessment is the best method for reducing the risk of drug-induced proarrhythmia in clinical practice. PMID- 16253927 TI - Use of preclinical assays to predict risk of drug-induced torsades de pointes. AB - Numerous medications, including drugs prescribed for noncardiac indications, can induce electrophysiologic changes that trigger the rare, malignant polymorphic ventricular tachyarrhythmia known as torsades de pointes (TdP). Although the exact relationship between electrophysiologic events and the development of TdP is not defined, prolongation of the QT interval and inhibition of the rapidly activating potassium current I(Kr) by drugs may be associated with an increased risk of TdP. The ability of a drug to reduce I(Kr) and prolong the QT interval often is considered to predict the likelihood that the drug will cause TdP in humans. However, these surrogate measures of the drug-induced risk of causing TdP, and therefore of drug safety, now are recognized to be imperfect predictors of drug safety. New preclinical models should be used to assess drug risk, including preparations, conditions, and measurements used by basic research scientists to produce ventricular polymorphic arrhythmias in the laboratory. In this review, we discuss the task of assessing the arrhythmogenic potential of a drug. Assays of drug effect to induce early afterdepolarizations and ectopic beats and/or to increase the dispersion of ventricular repolarization when "repolarization reserve" is reduced appear to be the best predictors of the drug induced risk of TdP. Current experimental models and protocols, especially those using conditions wherein the net repolarizing current is reduced, can detect the potential for a drug to induce TdP, even when the potential is extremely low. PMID- 16253928 TI - Clinical trial design to evaluate the effects of drugs on cardiac repolarization: current state of the art. AB - Prolongation of the QT interval associated with the potentially fatal arrhythmia known as torsades de pointes has been a common cause of the withdrawal of several promising drugs from the market. Many antihistamines, antibiotics, antimalarials, antidepressants, neuroleptics, antipsychotics, and imidazole antifungal agents have been shown to produce torsades, and all by the same mechanism. Advances in basic science and preclinical testing have begun to provide a scientific basis for distinguishing arrhythmogenicity from drug-induced QT effects. Many new techniques have been developed, and many others currently are being developed to facilitate the design of clinical trials to evaluate the effects of drugs on cardiac repolarization. The improvements in clinical trial design may help identify drugs that could induce torsades, halting futile research, potentially saving lives, and saving hundreds of millions of dollars in drug development. In the absence of any completely reliable surrogate measure for the arrhythmogenic potential of a drug, regulators have determined that QT interval prolongation should be intensively investigated in every drug that is developed. This article presents the basic mechanics of QT interval assessment and describes new developments that may make this measure a more accurate predictor of the effects of drugs on cardiac repolarization. It is absolutely essential that trial designs incorporate many ECG recordings, consistent QT interval measurement, and appropriate control or correction of the QT interval for heart rate in order to provide reproducible, scientifically meaningful results. PMID- 16253929 TI - Drug-induced torsades de pointes: the evolving role of pharmacogenetics. AB - Drug-induced torsades de pointes (TdP) is a rare, but potentially lethal, unwanted effect of drugs, including many commonly prescribed noncardiac drugs. Despite its low frequency, drug-induced TdP has generated a great deal of angst among physicians and pharmaceutical companies as well as tragedy, albeit rare, among patients. Although in retrospect many patients who died suddenly as a result of drug-induced TdP had identifiable risk factors, prediction in individual cases remains problematic. Over the past decade, tremendous progress has been made with respect to elucidating the fundamental pathogenic mechanisms that underlie drug-induced TdP. The vast majority of drugs associated with "QT liability" and the potential for drug-induced TdP, including all of the drugs removed from the market because of this side effect, are "HERG (human ether-a-go go-related gene) blockers." These drugs inhibit the KCNH2-encoded HERG potassium channel, which is one of the critical repolarizing forces involved in the exquisite orchestration of the heart's action potential. Consequently, myocyte repolarization is potentially delayed as evidenced by prolongation of the QT interval, thus providing the substrate for drug-induced TdP. Rare mutations in KCNH2 provide the pathogenic substrate for type 2 congenital long QT syndrome (LQTS), thus placing this cardiac potassium channel squarely in the intersection between congenital LQTS (the "Rosetta stone" of the heritable channelopathies) and acquired LQTS (drug-induced TdP). In addition, common polymorphisms residing in the LQTS-causing channel genes may confer heightened arrhythmogenic susceptibility and contribute to the makings of a vulnerable host. This review focuses on the present strategy of identifying "at-risk compounds" and the potential future strategy involving pharmacogenetics to pinpoint "at-risk hosts" in an effort to curb this rare, unintended, but potentially life-threatening side effect. PMID- 16253930 TI - Role of transmural dispersion of repolarization in the genesis of drug-induced torsades de pointes. AB - Torsades de pointes (TdP) is a potentially lethal arrhythmia that develops as a consequence of amplification of electrical heterogeneities intrinsic to the ventricular myocardium. These heterogeneities exist because of differences in the time course of repolarization of the three predominant cell types that make up the ventricular myocardium, giving rise to transmural voltage gradients and a dispersion of repolarization responsible for inscription of the ECG T wave. Antiarrhythmic agents with class III actions and/or the various mutations and cardiomyopathies associated with the long QT syndrome reduce net repolarizing current and amplify the intrinsic spatial dispersion of repolarization, thus creating the substrate for the development of reentry. The result is prolongation of the QT interval, abnormal T waves, and development of polymorphic reentrant ventricular tachycardia displaying characteristics of TdP. Prolongation of the QT interval apparently is not the sole determinant of a drug's potential to cause TdP. Agents that do not increase transmural dispersion of repolarization have little or no potential to induce TdP despite any ability to prolong the QT interval. In addition, drugs such as amiodarone and sodium pentobarbital can cause large QT prolongations but, by reducing transmural dispersion of repolarization, may reduce the likelihood of TdP. Arterially perfused wedge preparations of canine left ventricle can be used to explore the role of transmural dispersion of repolarization in the genesis of TdP. The purpose of this article is to review recent advances that have improved our understanding of these mechanisms, particularly the role of transmural dispersion of repolarization, in the genesis of drug-induced TdP and to examine how these advances can guide us toward the development of safer and more effective drugs. PMID- 16253931 TI - Tobacco outlet density, cigarette smoking prevalence, and demographics at the county level of analysis. AB - The geographic associations between tobacco outlet density, cigarette smoking prevalence, and demographic variables at the county unit of analysis were examined. End of year 2002 data were derived from licenses of 4745 tobacco selling retail outlets in all 99 Iowa counties. The 2000 census and the 2002 Iowa Behavioral Risk Factor Surveillance System (IBRFSS) were used to gather data for demographic variables and smoking prevalence rates. The IBRFSS telephone interviews were conducted from January through December in 2002 with a random sample of 3662 Iowa residents. As expected, results showed that counties with higher density of tobacco outlets and smoking prevalence also tended to have a higher percentage of minority residents. Contrary to previous studies, however, counties with higher tobacco outlet density and smoking prevalence also tended to have higher median household income. Findings are discussed in light of the state's low income inequality. Implications of the study and directions for future research are discussed. PMID- 16253932 TI - Adolescent anabolic steroid use, gender, physical activity, and other problem behaviors*. AB - To test the comparative value of strain theory and problem behavior theory as explanations of adolescent anabolic steroid use, this study examined gender specific relationships among steroid use, physical activity, and other problem behaviors. Based on the United States Centers for Disease Control and Prevention's 1997 Youth Risk Behavior Survey, a nationally representative sample of over 16,000 U.S. public and private high school students, binge drinking, cocaine use, fighting, and sexual risk-taking were associated with higher odds of lifetime steroid use. In gender-specific analyses, steroid use was strongly associated with female fighting and smokeless tobacco use as well as male sexual risk. Neither athletic participation nor strength conditioning predicted odds of steroid use after controlling for problem behaviors, nor did steroid-using athletes report more frequent use than steroid-using nonathletes. The study's limitations and policy implications were noted. These data suggest that other problem behaviors such as substance use, fighting, and sexual risk are better predictors of adolescent steroid use than physical activity. Interventions to prevent steroid use should not be limited to male participants in organized sports programs, but should also target adolescents identified as at risk for other problem behaviors. PMID- 16253933 TI - Street and working children of Delhi, India, misusing toluene: an ethnographic exploration. AB - Our qualitative study explored: the perceptions of street children indulging in whitener fluid misuse; the social, economic, and cultural determinants of use; and users' views regarding effective preventive and control strategies. Forty five in-depth interviews and three focus group discussions were conducted between March and December 2003. A purposive sample of those working children who were using toluene was selected by Snowball sampling. The paper discusses the: determinants of initiation and continued use; drug user social networks; psycho emotional deprivation and frustrations of these children; socio-cultural aspects like work driven need; others' attitudinal response towards them and their work; parental support or the lack of it; and strategies for prevention of this misuse. PMID- 16253934 TI - Injection risk behavior among women syringe exchangers in San Francisco. AB - Women who inject drugs in cities where syringe exchange programs (SEPs) are well established may have different risks for HIV infection. In 1997, we interviewed 149 female syringe exchangers in San Francisco, CA, a city with high rates of injection drug use that is home to one of the largest and oldest SEPs in the United States. In this report, we describe their sociodemographics, health, and risk behavior, and we examine factors associated with recent syringe sharing. Fifty percent of respondents were women of color and the median age was 38 years. Most (86%) injected heroin and nearly half were currently homeless or had recently been incarcerated. One-third of all women reported needle sharing in the prior month. This was higher than the rate of needle sharing reported by a mixed gender sample of San Francisco exchangers in 1993, although it resembled the rate reported by a mixed gender sample in 1992. In a multivariate analysis, syringe sharing was associated with age, housing status, and sexual partnerships. Syringe sharers were more likely to be young, homeless, or have a sexual partner who was also an injection drug user. While wide access to sterile syringes is an important strategy to reduce HIV transmission among injection drug users (IDU), syringe exchange alone cannot eradicate risky injection by female IDU. Additional efforts to reduce risky injection practices should focus on younger and homeless female IDU, as well as address selective risk taking between sexual partners. PMID- 16253935 TI - Emotional intelligence and acculturation to the United States: interactions on the perceived social consequences of smoking in early adolescents. AB - High emotional intelligence (EI) is associated with decreased adolescent smoking. Acculturation to the United States is a risk factor for adolescent smoking. High EI may buffer the relationship between acculturation to the United States and perceptions of the social consequences of smoking (PSC). Emotional intelligence is the ability to: accurately perceive, appraise, and express emotion; access and/or generate feelings in facilitating thought; understand emotion and emotional knowledge; and regulate emotions. Emotional intelligence (measured by the Multifactor Emotional Intelligence Scale, Adolescent Version), acculturation, and PSC were assessed in 2001 from 416 Southern California sixth graders (47% boys; mean age = 11.3 yrs; 32% Hispanic/ Latino, 29% Asian/Pacific Islander, 13% White, 19% Multiethnic, 6% Other). There was a significant EI x US acculturation interaction (p < 0.01) suggesting that those with high EI perceived more social consequences associated with smoking. As the U.S. population becomes increasingly diversified, identifying protective variables and designing effective prevention programs for adolescents of diverse ethnic and cultural backgrounds becomes important. PMID- 16253936 TI - Improving research transfer in the addictions field: a perspective from Canada. AB - This paper discusses how to enhance the research-to-practice exchange in the addictions field, while maintaining a balance with the demands and complexities of program delivery and policy development. It outlines the evolution of the concept of evidence-based practice, discusses the practical limitations and ways to improve transferring research to practice, and provides examples of research transfer activities in Canada. Practical limitations to research transfer include individual, organizational, and community factors. A strategic approach to research transfer includes addressing these limitations by combining dissemination activities with interventions such as individual instruction and incentives; building relationships among researchers, practitioners, and populations served; and obtaining commitments at a systemic level from funding bodies and research organizations to support research transfer. The potential is noted for the concept of workforce development to facilitate research transfer at organizational levels. The conclusion shown in this paper is that the tools and concept of evidence-based practice can lead the way to strengthening addictions programs and policies, and the development of a conceptual model for addiction research transfer in Canada would be a useful next step. PMID- 16253937 TI - Comparison of a population-based sample of "risky drinking" smokers and groups consuming just one substance. AB - This population-based study compares a group of "risky drinking" smokers (n = 137) with groups of either smokers (n = 760) or "risky drinkers" (n = 173) regarding sociodemographic, behavioral and motivational variables. The study took place in a rural and urban region of Germany in 1997. No differences were found in sociodemographics, nicotine-related variables, frequency of alcohol consumption, and most alcohol-related diagnoses. People who drink and smoke show a higher proportion of alcohol dependence (ES: h = .20), drink more alcohol per occasion (ES: d = .39), and are more motivated to decrease alcohol consumption (ES: h = .45). The data suggest focusing research in more alcohol-related clinical settings and examining the relationship between smoking cessation and alcohol reduction. PMID- 16253938 TI - Contrast effects in perceived risk of substance use. AB - Adolescent perceptions of the risks associated with the use of licit and illicit substances have important implications for policy and research. However, the methodological properties of the most popular risk measures in school surveys in Europe and the United States are not well understood. This study examines the potential contrast effects of risk measures of "heavy" and frequent substance use on perceived risks of occasional, moderate, and "experimental" use. Responses to 11 measures of the perceived risk of occasional smoking, moderate drinking, and experimental use of illicit substances were compared between two question forms administered to a split-half sample of all Icelandic ninth (14-15 years of age) and tenth (15-16 years of age) grade students present in class on the day of administration in March 2003 (N = 7099). In one form, only these 11 questions were used, while the other form also contained 13 questions on the perceived risk of heavy smoking, heavy drinking, and regular use of illicit substances. The longer form is found to decrease response rates and suppress estimates of perceived risk of experimental illicit substance use. Question form and perceived risks of heavy and regular use generally do not affect the multivariate effects of perceived risks of occasional, moderate, and experimental substance use on lifetime abstinence from each substance. It is argued that measures of perceived risks of heavy and regular substance use are less useful for prevention policy and research than are corresponding measures of occasional, moderate, and experimental substance use, and including the former in the same instrument may adversely affect the measurement of the latter constructs. PMID- 16253939 TI - Drug addiction in pregnancy and pregnancy outcome: a call for global solutions. PMID- 16253942 TI - Human responses to augmented virtual scaffolding models. AB - This study investigated the effect of adding real planks, in virtual scaffolding models of elevation, on human performance in a surround-screen virtual reality (SSVR) system. Twenty-four construction workers and 24 inexperienced controls performed walking tasks on real and virtual planks at three virtual heights (0, 6 m, 12 m) and two scaffolding-platform-width conditions (30, 60 cm). Gait patterns, walking instability measurements and cardiovascular reactivity were assessed. The results showed differences in human responses to real vs. virtual planks in walking patterns, instability score and heart-rate inter-beat intervals; it appeared that adding real planks in the SSVR virtual scaffolding model enhanced the quality of SSVR as a human - environment interface research tool. In addition, there were significant differences in performance between construction workers and the control group. The inexperienced participants were more unstable as compared to construction workers. Both groups increased their stride length with repetitions of the task, indicating a possibly confidence- or habit-related learning effect. The practical implications of this study are in the adoption of augmented virtual models of elevated construction environments for injury prevention research, and the development of programme for balance control training to reduce the risk of falls at elevation before workers enter a construction job. PMID- 16253943 TI - The development and validation of equations to predict grip force in the workplace: contributions of muscle activity and posture. AB - The inherent difficulty of measuring forces on the hand in ergonomic workplace assessments has led to the need for equations to predict grip force. A family of equations was developed, and validated, for the prediction of grip force using forearm electromyography (six finger and wrist muscles) as well as posture of the wrist (flexed, neutral and extended) and forearm (pronated, neutral, supinated). Inclusion of muscle activity was necessary to explain over 85% of the grip force variance and was further improved with wrist posture but not forearm posture. Posture itself had little predictive power without muscle activity (<1%). Nominal wrist posture improved predictive power more than the measured wrist angle. Inclusion of baseline muscle activity, the activity required to simply hold the grip dynamometer, greatly improved grip force predictions, especially at low force levels. While the complete model using six muscles and posture was the most accurate, the detailed validation and error analysis revealed that equations based on fewer components often resulted in a negligible reduction in predictive strength. Error was typically less than 10% under 50% of maximal grip force and around 15% over 50% of maximal grip force. This study presents detailed error analyses to both improve upon previous studies and to allow an educated decision to be made on which muscles to monitor depending on expected force levels, costs and error deemed acceptable by the potential user. PMID- 16253944 TI - Job and organizational determinants of nursing home employee commitment, job satisfaction and intent to turnover. AB - The purpose of this study was to examine whether job characteristics, the work environment, participation in quality improvement activities and facility quality improvement environment predicted employee commitment and job satisfaction in nursing homes, and whether those same predictors and commitment and satisfaction predicted turnover intention. A total of 6,584 nursing home employees from 76 nursing homes in a midwestern state participated. A self-administered questionnaire was used to collect the data. The results supported the hypotheses that job and organizational factors predicted commitment and satisfaction while commitment and satisfaction predicted turnover intentions. The implications for retaining nursing home employees are discussed. PMID- 16253945 TI - Shiftwork experience, age and cognitive performance. AB - Changes of alertness and cognitive efficiency has been suggested in people whose circadian rhythms are disrupted, e.g. night or shift-workers. Data from field and laboratory studies have demonstrated short-term cognitive disturbances related to circadian rhythm disruption. By contrast, little is known about the long-term consequences of chronic sleep deprivation, as can be observed with shift-work, on cognitive abilities. The present paper is aimed at evaluating, on a large cross sectional sample of workers, the long-term influence of shift-work on verbal memory and speed performances. Participants were 3,237 workers aged 32, 42, 52, and 62 years of various occupational statuses included in the VISAT (Aging, Health and Work) cohort. Data collected by questionnaires included items on working hours and shift-work and sleep disorders. Cognitive abilities were assessed using neuropsychological tests. Current male shift-workers had lower cognitive performance than never exposed workers. In the same population, memory performance tended to decrease with increasing shift-work duration. Among former shift-workers, the cognitive performance of the participant having stopped shiftwork more than 4 years ago seemed to be increased, suggesting a possible reversibility of effects. In conclusion, this study demonstrated that cognitive functioning tends to be impaired by a long-term exposure to SW. As found by other authors, neuropsychological performance tends to decrease with the increases in the duration of exposure to SW. PMID- 16253946 TI - Driver behaviour with adaptive cruise control. AB - This paper reports on the evaluation of adaptive cruise control (ACC) from a psychological perspective. It was anticipated that ACC would have an effect upon the psychology of driving, i.e. make the driver feel like they have less control, reduce the level of trust in the vehicle, make drivers less situationally aware, but workload might be reduced and driving might be less stressful. Drivers were asked to drive in a driving simulator under manual and ACC conditions. Analysis of variance techniques were used to determine the effects of workload (i.e. amount of traffic) and feedback (i.e. degree of information from the ACC system) on the psychological variables measured (i.e. locus of control, trust, workload, stress, mental models and situation awareness). The results showed that: locus of control and trust were unaffected by ACC, whereas situation awareness, workload and stress were reduced by ACC. Ways of improving situation awareness could include cues to help the driver predict vehicle trajectory and identify conflicts. PMID- 16253947 TI - Effect of concrete block weight and wall height on electromyographic activity and heart rate of masons. AB - Work-related musculoskeletal disorders (MSDs) are common among construction workers, such as masons. Few interventions are available to reduce masons' exposure to heavy lifting, a risk factor for MSDs. The purpose of this study was to determine whether one such intervention, the use of light-weight concrete blocks (LWBs), reduces physiological loads compared to standard-weight blocks (SWBs). Using a repeated measures design, 21 masons each constructed two 32-block walls, seven courses (rows) high, entirely of either SWBs or LWBs. Surface electromyography (EMG), from arm and back muscles, and heart rate was sampled. For certain muscles, EMG amplitudes were slightly lower when masons were laying LWBs compared to SWBs. Upper back and forearm extensor EMG amplitudes were greater for the higher wall courses for both block weights. There were no significant differences in heart rate between the two blocks. Interventions that address block weight and course height may be effective for masons. PMID- 16253949 TI - Hyaluronan: pharmaceutical characterization and drug delivery. AB - Hyaluronic acid (HA), is a polyanionic polysaccharide that consists of N-acetyl-D glucosamine and beta-glucoronic acid. It is most frequently referred to as hyaluronan because it exists in vivo as a polyanion and not in the protonated acid form. HA is distributed widely in vertebrates and presents as a component of the cell coat of many strains of bacteria. Initially the main functions of HA were believed to be mechanical as it has a protective, structure stabilizing and shock-absorbing role in the body. However, more recently the role of HA in the mediation of physiological functions via interaction with binding proteins and cell surface receptors including morphogenesis, regeneration, wound healing, and tumor invasion, as well as in the dynamic regulation of such interactions on cell signaling and behavior has been documented. The unique viscoelastic nature of hyaluronan along with its biocompatibility and nonimmunogenicity has led to its use in a number of cosmetic, medical, and pharmaceutical applications. More recently, HA has been investigated as a drug delivery agent for ophthalmic, nasal, pulmonary, parenteral, and dermal routes. The purpose of our review is to describe the physical, chemical, and biological properties of native HA together with how it can be produced and assayed along with a detailed analysis of its medical and pharmaceutical applications. PMID- 16253950 TI - Characteristics and biodistribution of soybean sterylglucoside and polyethylene glycol-modified cationic liposomes and their complexes with antisense oligodeoxynucleotide. AB - A novel cationic liposome modified with soybean sterylglucoside (SG) and polyethylene glycol-distearoylphosphatidylethanolamine (PEG-DSPE) as a carrier of antisense oligodeoxynucleotide (ODN) for hepatitis B virus (HBV) therapy was constructed. Characteristics of the cationic liposomes modified with SG and PEG (SG/PEG-CL) and their complexes with 15-mer phosphorothioate ODN (SG/PEG-CL-ODN complex) were investigated by incorporation efficiency, morphology, electrophoresis, zeta potentials, and size analysis. Antisense activity of the liposomes and ODN complexes was determined as hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg) in HepG2 2.2.15 cells by ELISA. Their tissue and intrahepatic distribution were evaluated following intravenous injection in mice. The complexes gained high incorporation efficiency and intact vesicular structure with mean size at approximately 200 nm. The SG/PEG-CL-ODN complexes enhanced the inhibition of both HBsAg and HBeAg expression in the cultured HepG2 2.2.15 cells relative to free ODN. The uptake of SG/PEG-CL and nonmodified cationic liposomes (CL) was primarily by liver, spleen, and lung. Furthermore, the concentration of SG/PEG-CL was significant higher than that of CL in hepatocytes at 0.5 hr postinjection. The biodistribution of SG/DSPE-CL-ODN complex compare with free ODN showed that liposomes enhanced the accumulation of ODN in the liver and spleen, while decreasing its blood concentration. SG/PEG-CL mediated ODN transfer to the liver is an effective gene delivery method for cell specific targeting, which has a potential for gene therapy of HBV infections. SG and PEG-modified cationic liposomes have proven to be an alternative carrier for hepatocyte-selective drug targeting. PMID- 16253951 TI - Evaluation of the interaction and drug release from alpha,beta-polyaspartamide derivatives to a biomembrane model. AB - This article reports on a comparative study on the ability of various polymers, containing hydrophilic and/or hydrophobic groups, to interact with a biomembrane model using the differential scanning calorimetry (DSC) technique. Multilamellar vesicles of mixed dimyristoylphosphatidylcholine (DMPC) and dimyristoylphosphatidic acid (DMPA) were chosen as a model of cell membranes. The investigated samples were a water soluble polymer, the alpha,beta-poly(N-2 hydroxyethyl)-DL-aspartamide (PHEA) and its derivatives partially functionalized with polyethylene glycol (PEG2000) to obtain PHEA-PEG2000, with hexadecylamine (C16) to obtain PHEA-C16, and with both compounds to obtain PHEA-PEG2000-C16. These polymers are potential candidates to prepare drug delivery systems. In particular, some samples give rise to polymeric micelles able to entrap hydrophobic drugs in an aqueous medium. The migration of drug molecules from these micelles to DMPC/DMPA vesicles also has been evaluated by DSC analysis, by using ketoprofen as a model drug. PMID- 16253952 TI - Bovine serum albumin-loaded pectinate beads as colonic peptide delivery system: preparation and in vitro characterization. AB - Objective of this study was to prepare a drug delivery system for therapeutic peptides that are degraded in the upper part of the gastrointestinal tract due to degradation activity of the enzymes. Delivering peptide to the colon in which enzymatic activity is low is next hope for absorption of these agents. Pectin, a naturally occurring water soluble polysaccharide, as a matrix for peptide delivery was studied. Degradation of pectin by the colonic enzymes makes it suitable for colon-specific delivery of drugs. Bovine serum albumin (BSA) was used as a model peptide. Calcium pectinate beads were prepared by extruding BSA loaded pectin solution to an agitating calcium chloride solution, and gelled spheres were formed instantaneously by an ionotropic gelation reaction. The effect of several factors such as concentration of pectin, concentration of calcium chloride, and total drug loading on the pattern of drug release in the dissolution medium was studied. Prepared beads showed good resistance in the release medium. The entrapment efficiency of the beads was high (between 63% and 99%). Entrapment efficiency of BSA was reversely dependent to the amount of the drug loaded in the beads. The amount of BSA loaded on the beads affects pattern of drug release. The concentration of the pectin showed the highest impact on the rate of drug release. Presence of the pectiolytic enzymes facilitated the drug release from the beads. PMID- 16253953 TI - Novel cationic copolymers of a polyasparthylhydrazide: synthesis and characterization. AB - Alpha,beta-poly(asparthylhydrazide) (PAHy), a water soluble synthetic polymer, was functionalized by using EDCI chemistry with 3-(carboxypropyl)trimethyl ammonium chloride (CPTACl) obtaining carboxypropyltrimethyl ammonium copolymers (PAHy-CPTA). Three PAHy-CPTA copolymers at increasing derivatization degrees (38%, 48%, 58%) were chosen for subsequent investigations. The capability of these copolymers to bind, neutralize, and protect DNA against degradation by DNase II was evalued by gel retardation assay and DNA degradation test at pH 5.5. Zeta potential measurements show that all studied polymers are able to neutralize the anionic charge of DNA at polymer/DNA weight ratio in the range of 0.8/1-5/1. Polyplex dimensional distribution analyses in bistilled water, saline solution NaCl 0.9%, and HEPES pH 7 show that polyplex size is strongly affected by both presence and type of electrolyte and with time incubation. PMID- 16253954 TI - Solid lipid nanoparticles containing tamoxifen characterization and in vitro antitumoral activity. AB - Solid lipid nanoparticles (SLNs) containing tamoxifen, a nonsteroidal antiestrogen used in breast cancer therapy, were prepared by microemulsion and precipitation techniques. Tamoxifen loaded SLNs seem to have dimensional properties useful for parenteral administration, and in vitro plasmatic drug release studies demonstrated that these systems are able to give a prolonged release of the drug in the intact form. Preliminary study of antiproliferative activity in vitro, carried out on MCF-7 cell line (human breast cancer cells), demonstrated that SLNs, containing tamoxifen showed an antitumoral activity comparable to free drug. The results of characterization studies and of in vitro antiproliferative activity strongly support the potential application of tamoxifen-loaded SLNs as a carrier system at prolonged release useful for intravenous administration in breast cancer therapy. PMID- 16253955 TI - Effect of co-solvents on the controlled release of calcitonin polypeptide from in situ biodegradable polymer implants. AB - The objective of this study was to design an in situ biodegradable polymer implant controlled-release drug delivery system, using novel combinations of co solvents and a model polypeptide, calcitonin (CT), and to assess the release of drug as a function of these co-solvents. Formulations were prepared by dissolving/ suspending CT polypeptide in poly-(lactic acid) (PLA) polymer solutions/suspensions containing combinations of a hydrophobic (benzyl benzoate, BB) and a hydrophilic (benzyl alcohol, BA) solvent. The CT-PLA mixtures were each injected into test tubes containing phosphate buffered saline solution to form the in situ implant and sampling was conducted over a 28-day period. The samples were analyzed for drug content using a modified Lowry protein assay procedure. Cumulative drug release demonstrated a rank-order correlation depending on the amount of the hydrophobic (BB) and hydrophilic (BA) solvents within each system. Increasing the amounts of the hydrophobic solvent, BB, in formulations demonstrated a 1.2-4.4-fold increase in CT release. Stability studies of all formulations over a 4-month period showed progressive increase in degradation of the CT polypeptide, especially at 37 degrees C, but a slower degradation pattern prevailed at 4 degrees and 20 degrees C. Differential scanning calorimetric studies revealed a homogenous mixture of drug in the polymer matrix. Overall, these studies demonstrated the feasibility of designing controlled release systems capable of releasing a polypeptide drug as a function of influence of different co-solvent combinations. PMID- 16253956 TI - Niosomes with sorbitan monoester as a carrier for vaginal delivery of insulin: studies in rats. AB - To prepare and investigate the potential of the niosomes vaginal delivery system for systemic treatment of insulin is the goal of this study. Two kinds of vesicles with Span 40 and Span 60 were prepared by lipid phase evaporation methods with sonication. The niosomal entrapment efficiency was determined by column chromatography. The particle size and morphology of the vesicles also were evaluated. The results showed optimized niosomes prepared in this study had niosomal entrapment efficiency 26.68 +/- 1.41% for Span 40 and 28.82 +/- 1.35% for Span 60, respectively. The particle sizes of Span 40 niosomes and Span 60 niosomes were 242.5 +/- 20.5 nm and 259.7 +/- 33.8 nm, respectively. There were no significant differences in appearance between the two types of vesicles. The hypoglycemic effects, and insulin concentrations after vaginal administration of insulin vesicles to rats were investigated. Compared with subcutaneous administration of insulin solution, the relative pharmacological bioavailability and the relative bioavailability of vaginal administration of insulin vesicles were determined. Compared with subcutaneous administration of insulin solution, the relative pharmacological bioavailability and the relative bioavailability of insulin-Span 60 vesicles group were 8.43% and 9.61%, and insulin-Span 40 niosomes were 9.11% and 10.03% (p > 0.05). Span 60 and Span 40 niosomes were both higher than blank Span 40, Span 60 vesicles, and free insulin physical mixture groups (p < 0.05). The results indicates insulin-Span 60, Span 40 niosomes had an enhancing effect on vaginal delivery of insulin. Although the factors controlling the process for penetration of a portion of vaginally administrated niosomes into bloodstream from vaginal tract is still not fully understood, our results demonstrated that after encapsulation in niosomes of definite type, insulin became an active and efficiently therapeutic agent when administrated vaginally and might be a good carrier for vaginal delivery of protein drugs. PMID- 16253957 TI - Encapsulation and in vitro evaluation of amikacin-loaded erythrocytes. AB - The aim of our present work was to establish the effect of the osmolality of the hypotonic buffer on the encapsulated amount and the in vitro properties of Amikacin-loaded erythrocytes. Amikacin was encapsulated in rat erythrocytes using a hypotonic dialysis method with hypotonic buffers of different osmolalities with mean values around 90 and 150 mOsm/kg. Morphological examination of the ghost erythrocytes was accomplished using scanning electron microscopy (SEM). The osmotic fragility of normal and loaded erythrocytes was tested using hypotonic solutions. Evaluation of the hematological parameters of the control and loaded erythrocytes was carried out using a hematology system analyzer. Amikacin release from loaded erythrocytes was tested in autologous plasma at 37 degrees C over a 24-h period. The quantification of Amikacin in loaded erythrocytes and in autologous plasma was performed using an HPLC technique. A higher osmotic fragility of loaded erythrocytes was observed using a low osmolality buffer. Some hematological parameters showed statistically significant differences between the loaded erythrocytes obtained using two buffers of different osmolalities with respect to untreated erythrocytes. According to our results, Amikacin carrier erythrocytes obtained by hypotonic dialysis using a low osmolality buffer (90 mOsm/kg) should afford a good encapsulation yield, appropriate morphological properties, and sustained release in vitro. PMID- 16253958 TI - A role of p44/42 mitogen-activated protein kinases in formyl-peptide receptor mediated phospholipase D activity and oxidant production. AB - Phosphatidylcholine-specific phospholipase D (PLD) is a major cellular source of phosphatidic acid and choline, which regulate various physiopathological processes. PLD activation mediated by chemoattractants involves protein phosphorylation. This study provides pharmacological and biochemical evidence of a major role of p44/42 MAP kinases (ERK1/2) in PLD activation induced by the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine (fMLP). ERK1/2 inhibition by the MEK1/2 antagonist U0126 in neutrophilic HL-60 cells or HEK 293T cells stably expressing fMLP receptors abolished fMLP-mediated PLD activity. Conversely, a constitutively activated MEK1 mutant expressed in HEK 293T cells potentiated fMLP-induced PLD activity. Expression of inactive PLD mutants showed that PLD2, but not PLD1, contributed to fMLP-mediated PLD activity. PLD2 co immunoprecipitated with ERK1/2 and became phosphorylated on MAP kinase consensus sites in fMLP-stimulated cells. In cell-free systems, ERK2 gave rise to strong ATP-dependent PLD activity and directly phosphorylated PLD2 that generated two phosphopeptides only after tryptic digestion. Finally, pharmacological inhibition of ERK activation and the inhibition of PLD expression by antisense oligonucleotides in HL-60 cells suggest that the ERK/PLD2 pathway contributes to fMLP-mediated oxidant production. In conclusion, the fMLP-mediated PLD activity is regulated by ERK1/2, involving a predominant contribution of PLD2. The ERK/PLD2 coupling may provide potential pharmacological targets to control PLD associated cellular dysfunctions. PMID- 16253959 TI - Crystal structure of the E230Q mutant of cAMP-dependent protein kinase reveals an unexpected apoenzyme conformation and an extended N-terminal A helix. AB - Glu230, one of the acidic residues that cluster around the active site of the catalytic subunit of cAMP-dependent protein kinase, plays an important role in substrate recognition. Specifically, its side chain forms a direct salt-bridge interaction with the substrate's P-2 Arg. Previous studies showed that mutation of Glu230 to Gln (E230Q) caused significant decreases not only in substrate binding but also in the rate of phosphoryl transfer. To better understand the importance of Glu230 for structure and function, we solved the crystal structure of the E230Q mutant at 2.8 A resolution. Surprisingly, the mutant preferred an open conformation with no bound ligands observed, even though the crystals were grown in the presence of MgATP and the inhibitor peptide, IP20. This is in contrast to the wild-type protein that, under the same conditions, prefers the closed conformation of a ternary complex. The structure highlights the importance of the electrostatic surface not only for substrate binding and catalysis, but also for the mechanism for closing the active site cleft. This surface mutation clearly disrupts the recognition and binding of substrate peptide so that the enzyme prefers an open conformation that cannot trap ATP. This is consistent with the reinforcing concepts of conformational dynamics and the synergistic binding of ATP and substrate peptide. Another unusual feature of the structure is the observation of the entire N terminus (Gly1-Thr32) assumes an extended alpha-helix conformation. Finally, based on temperature factors, this mutant structure is more stable than the wild-type C-subunit in the apo state. PMID- 16253960 TI - Dietary (n-3) long chain polyunsaturated fatty acids prevent sucrose-induced insulin resistance in rats. AB - This study was designed to determine the effect of substituting (n-3) long-chain PUFAs (LCPUFAs) for linoleic acid and hence decreasing the (n-6):(n-3) fatty acid ratio on sucrose-induced insulin resistance in rats. Weanling male Wistar rats were fed casein-based diets containing 100 g/kg fat for 12 wk. Insulin resistance was induced by replacing starch (ST) with sucrose (SU). The dietary fats were formulated with groundnut oil, palmolein, and fish oil to provide the following ratios of (n-6):(n-3) fatty acids: 210 (ST-210, SU-210), 50 (SU-50), 10 (SU-10), and 5 (SU-5). Compared with starch (ST-210), sucrose feeding (SU-210) significantly increased the plasma insulin and triglyceride concentrations and the plasma insulin area under the curve (AUC) in response to an oral glucose load. Adipocytes isolated from rats fed SU-210 had greater lipolytic rate, lower insulin stimulated glucose transport, and lower insulin-mediated antilipolysis than those from rats fed ST-210. Decreasing the dietary (n-6):(n-3) ratio in sucrose-fed rats (SU-10 and SU-5) normalized the plasma insulin concentration and the AUC of insulin after a glucose load. The sucrose-induced increase in plasma triglyceride concentration was normalized in rats fed SU-50, SU-10 and SU-5. Further, sucrose-induced alterations in adipocyte lipolysis and antilipolysis were partially reversed and glucose transport improved in rats fed diets SU-5 and SU-10. In diaphragm phospholipids, decreasing the (n-6):(n-3) ratio in the diet increased the concentration of (n-3) LCPUFAs with concomitant decreases in the concentration of (n-6) LCPUFAs. These results suggest that (n-3) LCPUFAs at a level of 2.6 g/kg diet [0.56% energy (n-3) LCPUFAs, (n-6):(n-3) ratio = 10] may prevent sucrose-induced insulin resistance by improving peripheral insulin sensitivity. PMID- 16253961 TI - The post-menopausal ovary displays a unique pattern of steroidogenic enzyme expression. AB - BACKGROUND: While menopause results in the loss of cyclic steroid production, evidence exists for persistent, albeit reduced, ovarian androgen production. In order to continue to synthesize ovarian androgens, the steroidogenic enzymes necessary for androgen biosynthesis must be present. Few studies have selectively analysed some of the steroidogenic enzymes present in the post-menopausal ovary (PMO), and a comprehensive study of this matter has never been undertaken. METHODS: RNA and protein were obtained from PMO, pre-menopausal ovarian stroma, corpora lutea (CL), ovarian follicles, placenta, and myometrium. Oligonucleotide microarray analysis was performed to compare the gene expression profiles of PMO with pre-menopausal ovarian stroma. Real-time RT-PCR was performed for LH/HCG receptor (LHCGR), steroidogenic acute regulatory (StAR), cholesterol side-chain cleavage (CYP11A), 3beta-hydroxysteroid dehydrogenase type I (HSD3B1) and type II (HSD3B2, 3betaHSD), 17a-hydroxylase (CYP17), cytochrome b5 (CytB5), and aromatase (CYP19). Western blot analysis was performed for StAR, CYP11A, CYP17,and 3betaHSD. RESULTS: The PMO and pre-menopausal ovarian stroma had a similar pattern of steroidogenic enzyme expression. The PMO had persistent, but reduced, levels of LHCGR and most steroidogenic enzymes. CYP19 and HSD3B2 mRNA were greatly reduced in PMO in comparison with CL (50-fold and 2000-fold less respectively). HSD3B2 was not detectable in PMO by western analysis. CONCLUSIONS: This study supports the idea that the PMO retains some steroidogenic capacity. However, based on steroidogenic enzyme expression, the PMO has a unique pattern of steroidogenic enzyme expression that favors Delta5 steroid formation over Delta4 steroid formation. PMID- 16253962 TI - Tumour necrosis factor-alpha, interleukin-6 and interleukin-8 do not promote adhesion of human endometrial epithelial cells to mesothelial cells in a quantitative in vitro model. AB - BACKGROUND: A key factor in the pathogenesis of endometriosis is the endometrial peritoneal adhesion. To study the pathogenesis of endometriosis, a quantitative in vitro assay (QIVA) was developed to measure in vitro adhesion between human endometrial epithelial cells and mesothelial cells using commercially available cell lines. Using the QIVA, the hypothesis was tested that tumour necrosis factor alpha (TNF-alpha), interleukin-6 (IL-6) and interleukin-8 (IL-8) promote adhesion of endometrial epithelial cells to mesothelial cells. METHODS: Mesothelial cells were pre-treated with TNF-alpha, IL-6 or IL-8 in various concentrations (ranging from 0 to 1000 IU/ml) for 24 h. Confluent endometrial epithelial cells were labelled with [35S]methionine, added to the confluent mesothelial cells and incubated for 1 h. After incubation, non-adhering cells were removed and adherent cells were solubilized and their [35S]methionine radioactivity was counted to quantify the adherence of endometrial epithelial cells to mesothelial cells. RESULTS: The in vitro adhesion of human endometrial epithelial cells to human mesothelial cells was inhibited in a dose-dependent manner by TNF-alpha (P=0.0007), IL-6 (P<0.0001) and IL-8 (P=0.0004). CONCLUSIONS: Using a quantitative in vitro adhesion assay, we were unable to confirm our hypothesis that TNF-alpha, IL-6 and IL-8 promote the in vitro adhesion between endometrial epithelial cells and mesothelial cells. PMID- 16253963 TI - Expression of human serum albumin (HSA) mRNA in human granulosa cells: potential correlation of the 95 amino acid long carboxyl terminal of HSA to gonadotrophin surge-attenuating factor. AB - INTRODUCTION: According to previous studies, gonadotrophin surge-attenuating factor (GnSAF), which is assumed to be produced in human granulosa cells, has a homology with the carboxyl terminal of the human serum albumin (HSA) protein. In an attempt to validate these findings, whole or partial expression of the HSA gene was studied by RT-PCR analysis in human granulosa cells from women undergoing IVF treatment. METHODS: RT-PCR analysis of HSA RNA transcripts in luteinized granulosa cells was done in order to investigate the possible expression of the HSA gene. To ensure the specificity of PCR products, restriction enzyme and sequence analysis were performed. Western blot analysis was carried out to detect the possible expression of the albumin gene in granulosa cells. RESULTS: RT-PCR analysis and sequencing analysis of cDNA from granulosa cells revealed bands identical with those from the positive control for the amino as well as the carboxyl terminal corresponding to HSA gene at the cytoplasmic level. CONCLUSION: We have demonstrated that human granulosa cells express the carboxyl and amino terminal part of the HSA gene in levels comparable to those found in human hepatocytes. It is suggested that the coding gene for GnSAF may be a result of an alternative expression of HSA gene. PMID- 16253964 TI - Drug-induced apoptosis was markedly attenuated in endometriotic stromal cells. AB - BACKGROUND: The survival of endometriotic cells in the ectopic site has been investigated from the aspect of susceptibility of endometriotic tissues to apoptosis. In order to investigate the nature of abnormal survival of endometriotic cells in ectopic locations, we compared drug-induced apoptosis in endometrial and endometriotic cells. METHODS: Endometrial stromal cells were obtained from normal endometrium in 11 patients who underwent hysterectomy for leiomyoma without endometriosis. Endometriotic cells were isolated from the chocolate cyst linings of the ovary in 13 patients who underwent laparoscopic surgery. Cells were cultured in the presence or absence of staurosporine. Apoptotic cell death was evaluated by staining nuclei with propidium iodide and phosphatidylserine (a marker of early apoptotic events) with Annexin V as well as by DNA fragmentation assay. The number of viable cells was estimated by modified MTT [3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide WST-8] assay. RESULTS: After 3 h of exposure to staurosporine, >50% of the endometrial stromal cells became Annexin V positive. In contrast, >30% of the endometriotic cells were Annexin V positive. DNA fragmentation was not clearly induced in the endometriotic cells. Less than 20% of the endometrial cells survived after staurosporine exposure, while >40% of the endometriotic cells survived. Cell death induced by staurosporine was partially blocked by incubation with the caspase inhibitor, N-benzyoxycarbonyl-Val-Ala-Asp(OMe)fluoromethyl-ketone (ZVAD fmk), suggesting that a caspase cascade may play a role in the cell death process. CONCLUSIONS: Attenuated susceptibility to apoptosis in endometriotic stromal cells may be associated with abnormal survival in ectopic sites in an environment that is probably unfavourable. These results may be implicated in the pathophysiology of endometriosis. PMID- 16253965 TI - Ovarian cyst formation following GnRH agonist administration in IVF cycles: incidence and impact. AB - BACKGROUND: The formation of functional ovarian cysts has been recognized as one of the side effects of GnRH agonist administration. The formation of cysts during IVF treatment may be of no clinical significance or may negatively influence its outcome. The objective of this study was to determine the incidence of ovarian cyst formation following GnRH agonist administration and to examine their effect on IVF outcome. METHODS: A prospective study of 1317 IVF patients who developed one or more functional ovarian cysts of >or=15 mm following GnRH agonist treatment was performed. Transvaginal ultrasonographic-guided cyst aspiration was carried out in 76 randomly allocated patients out of 122 patients who were found to have functional ovarian cysts before starting ovarian stimulation with gonadotropins. RESULTS: The incidence of follicular cyst formation was 9.3%. Cyst cycles in comparison with non-cyst cycles had significantly elevated day 3 basal FSH (mean+/-SD of 8.3+/-3.2 versus 5.3+/-2.6 mIU/ml, P<0.05) and required more ampoules of gonadotropins (46.3+/-16.5 versus 35+/-14.6, P<0.01). Furthermore, they showed a statistically significant decrease in the quality and number of oocytes retrieved, fertilization rate, number and quality of embryos, implantation and pregnancy rates, with a significant increase in cancellation and abortion rates. Patients with bilateral cysts had a significantly lower number of oocytes and embryos retrieved, with a lower proportion of metaphase II oocytes. They also had a higher proportion of poor quality embryos. Cyst aspiration was not associated with a significant difference in the above parameters. CONCLUSIONS: The incidence of cyst formation during GnRH agonist treatment is lower than previously reported. In such cases, the quality of oocytes and embryos were significantly compromised, with a significant increase in the cycle cancellation rate and a decrease in the implantation and pregnancy rates. Neither conservative management nor cyst aspiration improved the IVF outcome. PMID- 16253966 TI - Embryogenesis of chimeras, twins and anterior midline asymmetries. AB - Human spontaneous chimerism, with one body built from cells of both twins of a dizygotic (DZ) pair, is supposed to be extremely rare, arising from the exchange of blood cells through placental anastomoses. Mosaicism is supposed to be far more common, arising from single zygotes by embryonic mutation. Because typical diagnosis of mosaicism can neither identify nor exclude chimerism, 'mosaicism' may often be chimerism undiscovered. Evidence shows chimerism arises primarily from DZ embryo fusion and is not rare, although it has negligible probability under the hypothesis of independent double ovulation and independent embryogenesis. If, instead, DZ twin embryos begin development as a single cell mass, chimerism is likely. This would be consistent with observations that DZ twins develop as differently from singletons as monozygotic twins do with regard to embryogenic establishment of asymmetries of midline neural-crest-driven structures of brain, face and heart. Chimerism is a significant component of human embryonic development that deserves closer attention as a mechanism of developmental variation. The 'common knowledge' understanding of twinning mechanisms is at best inadequate. The importance of the difference lies in what we can learn from chimerism about human embryogenesis and the cellular origins of structures and functions basic to the business of becoming human. PMID- 16253967 TI - Antiphospholipid antibodies in serum and follicular fluid--is there a correlation with IVF implantation failure? AB - BACKGROUND: Antiphospholipid antibodies (aPLs) are associated with infertility, but the mechanism underlying this statistical association is currently obscure. We aimed to investigate the finding that aPLs are concentrated in follicular fluid and to establish if this is associated with a poorer outcome from IVF. METHODS AND RESULTS: In 19.2% of 99 women undergoing IVF, at least one aPL was detected in their serum and/or follicular fluids, but the antibody levels in follicular fluid were not higher than in serum. Women with aPLs had a lower implantation rate (14%) than women without these antibodies (24.1%), but this difference was not significant (P=0.127). There was also a non-significant reduction in the live birth rate for women with aPLs. In a parallel investigation, 10 sheep immunized with beta2 glycoprotein I (beta2GPI) or irrelevant control antigens showed strong immune responses, but there were no significant differences between the levels of antibodies in the follicular fluid or serum from beta2GPI or control immunized sheep. CONCLUSION: aPLs do not appear to be selectively concentrated in follicular fluids and, when present, do not adversely affect the reproductive outcome of women undergoing IVF. PMID- 16253968 TI - High density of small nerve fibres in the functional layer of the endometrium in women with endometriosis. AB - BACKGROUND: Endometriosis is a common gynaecological disease and is frequently associated with recurrent and serious pelvic pain such as dysmenorrhoea and dyspareunia, but the mechanisms by which these symptoms are generated are not well understood. METHODS: Histological sections of endometrial tissue were prepared from endometrial curettings and hysterectomies performed on women with endometriosis (n=25 and n=10, respectively) and without endometriosis (n=47 and n=35, respectively). These were stained immunohistochemically for the highly specific polyclonal rabbit anti-protein gene product 9.5 (PGP9.5) and monoclonal mouse anti-neurofilament protein (NF) to demonstrate both myelinated and unmyelinated nerve fibres. RESULTS: Small nerve fibres were identified throughout the basal and functional layers of the endometrium in all endometriosis patients, but were not seen in the functional layer of the endometrium in any of the women without endometriosis (P<0.001). NF-immunoreactive nerve fibres were present in the basal layer in all endometriosis patients but not in non-endometriosis patients, with one exception (P<0.001). CONCLUSIONS: Small nerve fibres detected in the functional layer in all women with endometriosis may have important implications for understanding the generation of pain in these patients. The presence of nerve fibres in an endometrial biopsy may be a novel surrogate marker of clinical endometriosis. PMID- 16253970 TI - Is controlled ovarian stimulation in intrauterine insemination an acceptable therapy in couples with unexplained non-conception in the perspective of multiple pregnancies? AB - BACKGROUND: Controlled ovarian stimulation (COS) with intrauterine insemination (IUI) is a common treatment in couples with unexplained non-conception. Induction of multifollicular growth is considered to improve pregnancy outcome, but it contains an increased risk of multiple pregnancies and ovarian hyperstimulation syndrome. In this study the impact of the number of follicles (>14 mm) on the ongoing pregnancy rate (PR) and multiple PR was evaluated in the first four treatment cycles. METHODS: A retrospective cohort study was performed in all couples with unexplained non-conception undergoing COS-IUI in the Academic Hospital of Maastricht. The main outcome measure was ongoing PR. Secondary outcomes were ongoing multiple PR, number of follicles of >or=14 mm, and order of treatment cycle. RESULTS: Three hundred couples were included. No significant difference was found in ongoing PR between women with one, two, three or four follicles respectively (P=0.54), but in women with two or more follicles 12/73 pregnancies were multiples. Ongoing PR was highest in the first treatment cycle and declined significantly with increasing cycle order (P=0.006), while multiple PR did not change. CONCLUSIONS: In COS-IUI for unexplained non-conception, induction of more than one follicle did not improve the ongoing PR, but increased the risk of multiple pregnancies. Multiple PR remained high in the first four cycles with multifollicular stimulation. Therefore, in order to reduce the number of multiple pregnancies, in all IUI cycles for unexplained non-conception monofollicular growth should be aimed at. PMID- 16253969 TI - Maternal CD46H*2 and IL1B-511*1 homozygosity in T helper 1-type immunity to trophoblast antigens in recurrent pregnancy loss. AB - BACKGROUND: Women with recurrent pregnancy loss (RPL) and T-helper (Th)1-type immunity to trophoblast antigens have an increased frequency of the IL1B-511*1 promoter variant. Since CD46 gene products also regulate maternal immune responses including Th1 immunity, we investigated whether CD46 gene polymorphisms are also associated with RPL in women with and without Th1 immunity to trophoblast, and the possibility of a synergistic effect with the IL1B-511*1 promoter variant. METHODS: A case-controlled study was performed to document HindIII site polymorphism in intron 1 of the CD46 gene in 131 women with RPL and 72 fertile controls. Clinical information, Th1-type immune responsiveness to trophoblast in women with RPL history, and IL1B promoter allelotypes for this cohort were documented in a previous study. RESULTS: The frequency of the CD46H*2 allele and CD46H*2 homozygosity were significantly increased in women with RPL compared with fertile controls (P<0.028 and P<0.011). CD46H*2 homozygosity was highly associated with RPL-Th1(+) (32.4 versus 9.7% in fertile controls, P<0.0045). Logistic regression analysis revealed that women homozygous for both the IL1B-511*1 and CD46H*2 alleles had an extremely high risk of RPL-Th1(+) [exponential coefficients (EC)=24]. Among women with RPL, homozygosity at both alleles, but not each alone, significantly increased the risk of Th1 immunity to trophoblast antigens (EC=16), suggesting a possible genetic interaction between these two alleles in the development of Th1 immunity. CONCLUSIONS: The combination of homozygosity for both IL1B-511*1 and CD46H*2 alleles is a high risk factor for RPL-Th1(+). PMID- 16253971 TI - Hyper-osmotic condition enhances protein tyrosine phosphorylation and zona pellucida binding capacity of human sperm. AB - BACKGROUND: The aim of this study was to determine the effect of culture medium osmolality, in the range known to occur in the male and female reproductive tracts, on human sperm tyrosine phosphorylation and sperm-zona pellucida (ZP) interaction in vitro. METHODS: Motile sperm (2x10(6)), selected by swim-up from semen of normozoospermic men with normal sperm-ZP binding, were incubated with or without four oocytes in 1 ml human tubal fluid (HTF) medium with different osmolalities (150, 200, 280, 350, 400 mOsm/kg) adjusted by variation of the NaCl concentration. After 2 h incubation, the number of sperm bound to the four ZP was examined, sperm motility and velocities were assessed by Hamilton-Thorn Motility Analyzer (IVOS 10) and sperm tyrosine phosphorylation was assessed by both western immunoblotting and immunofluorescence with an anti-phosphotyrosine monoclonal antibody (PY20). The effect of hyper-osmolality (400 mOsm/kg) on the ZP-induced acrosome reaction (AR) was also determined. RESULTS: Incubation of human sperm in hyper-osmotic medium significantly increased tyrosine phosphorylation and the number of sperm bound to the ZP. In contrast, hypo osmotic medium significantly decreased both tyrosine phosphorylation and sperm-ZP binding. Medium with high osmolality (400 mOsm/kg) significantly reduced the ZP induced AR. Both hypo- and hyper-osmotic media significantly decreased average sperm percentage progressive motility and velocities. CONCLUSION: Incubation of human sperm in hyper-osmotic media was associated with significantly increased tyrosine phosphorylation and ZP-binding ability but severely reduced the ZP induced AR. PMID- 16253972 TI - Trends in embryo transfer practices and multiple gestation for IVF procedures in the USA, 1996-2002. AB - BACKGROUND: Increasing use of IVF in the USA has been a major contributor to the rising national multiple birth rate. Many have advocated that reducing the number of embryos transferred is essential for addressing the IVF-associated multiple birth problem. METHODS: A population-based sample of 506 072 IVF transfers performed in the USA in 1996-2002 was used to investigate trends in embryo transfer practices and to determine whether any changes in practice patterns have impacted the multiple gestation risk associated with IVF. RESULTS: The proportion of procedures in which >or=3 embryos were transferred declined significantly for most patient groups between 1996 and 2002. However, declines for some groups were not sizeable (from 79 to 73% and from 76 to 71% for fresh, non-donor procedures among women aged 38-40 and 41-42 years respectively) and transferring >or=3 embryos remained the norm for all groups. As of 2002, single embryo transfer had not increased for most groups and remained uncommon. Some declines in overall multiple gestation rates were observed, although multiple gestation risk associated with 2 embryos transferred increased significantly for all groups. CONCLUSIONS: Despite changes in embryo transfer practices, multiple gestation risk remains high, in part due to increased multiple gestation rates associated with the transfer of two embryos. PMID- 16253973 TI - In unselected patients, elective single embryo transfer prevents all multiples, but results in significantly lower pregnancy rates compared with double embryo transfer: a randomized controlled trial. AB - BACKGROUND: Elective single embryo transfer (eSET) in a selected group of patients (i.e. young patients with at least one good quality embryo) reduces the number of multiple pregnancies in an IVF programme. However, the reduced overall multiple pregnancy rate (PR) is still unacceptably high. Therefore, a randomized controlled trial (RCT) was conducted comparing eSET and double embryo transfer (DET) in an unselected group of patients (i.e. irrespective of the woman's age or embryo quality). METHODS: Consenting unselected patients were randomized between eSET (RCT-eSET) (n = 154) or DET (RCT-DET) (n = 154). Randomization was performed just prior to the first embryo transfer, provided that at least two 2PN zygotes were available. Non-participants received our standard transfer policy [SP-eSET in a selected group of patients (n = 100), otherwise SP-DET (n = 122)]. RESULTS: The ongoing PR after RCT-eSET was significantly lower as compared with RCT-DET (21.4 versus 40.3%) and the twin PR was reduced from 21.0% after RCT-DET to 0% after RCT-eSET. The ongoing PRs after SP-eSET and SP-DET did not differ significantly (33.0 versus 30.3%), with an overall twin PR of 12.9%. CONCLUSION: To avoid twin pregnancies resulting from an IVF treatment, eSET should be applied in all patients. The consequence would be a halving of the ongoing PR as compared with applying a DET policy in all patients. The transfer of one embryo in a selected group of good prognosis patients leads to a less drastic reduction in PR but maintains a twin PR of 12.9%. PMID- 16253974 TI - A prospective study of the forearm bone density of users of etonorgestrel- and levonorgestrel-releasing contraceptive implants. AB - BACKGROUND: The aim of the study was to compare bone mineral density (BMD) before insertion and at 18 months of use of etonorgestrel- and levonorgestrel-releasing contraceptive implants. METHODS: One hundred and eleven women, 19-43 years of age, were randomly allocated to two groups: 56 to etonorgestrel and 55 to levonorgestrel. BMD was evaluated at the midshaft of the ulna and at the distal radius of the non-dominant forearm using dual-energy X-ray absorptiometry before insertion and at 18 months of use. RESULTS: There was no difference in baseline demographic or anthropometric characteristics, or in BMD of users of either model of implant. BMD was significantly lower at 18 months of use at the midshaft of the ulna in both groups of users. However, no difference was found at the distal radius. Multiple linear regression analysis showed that the variables associated with BMD at 18 months of use in both implant groups were baseline BMD, body mass index (BMI) and difference in BMI (0 versus 18 months of use). CONCLUSIONS: Women of 19-43 years of age using either one of the implants showed lower BMD at 18 months of use at the midshaft of the ulna, however, without a difference at the distal radius. PMID- 16253975 TI - Uterine effects of metformin administration in anovulatory women with polycystic ovary syndrome. AB - BACKGROUND: Metformin has been shown to improve fertility in anovulatory patients with polycystic ovary syndrome (PCOS), inducing not only a high ovulation and pregnancy rate but also reducing the incidence of miscarriages. The aim of the present study was to evaluate the uterine effects of metformin in patients with PCOS who ovulated under metformin. METHODS: Thirty-seven non-obese primary infertile anovulatory patients with PCOS and another 30 age- and body mass index matched healthy women (control group) were studied. PCOS patients were treated with metformin (850 mg twice daily) for 6 months, whereas the control group did not receive any treatment. In these PCOS patients who ovulated whilst under metformin treatment (PCOS group) and in controls, uterine, sub-endometrial and endometrial blood flow, and endometrial thickness and pattern were evaluated using serial ultrasonographic assessments. RESULTS: Before treatment, uterine, sub-endometrial and endometrial blood flows were significantly lower in patients with PCOS than in the control group. All indexes of uterine vascularization were significantly improved in the PCOS group with metformin treatment and were not different from the controls. Nor was any difference in endometrial thickness and pattern detected between PCOS and control groups. After grouping the data of PCOS patients who ovulated under metformin for cycles with favourable/unfavourable reproductive outcome, no difference in any parameter was observed. CONCLUSIONS: Metformin improves all surrogate markers of endometrial receptivity in PCOS patients, without difference between patients who had favourable or unfavourable reproductive outcome. PMID- 16253976 TI - Rejection patterns in allogeneic uterus transplantation in the mouse. AB - BACKGROUND: Transplantation of the uterus in the mouse has been developed as a model system for research towards human uterine transplantation. Previous studies in a mouse model have demonstrated that a syngeneic uterus transplant can give rise to normal offspring. The aim of this study was to characterize the time course of rejection in a fully allogeneic mouse uterus transplantation model. METHODS: Uteri of BALB/c mice were transplanted to a heterotopic position in C57BL/6 recipients, whose native uteri were left in situ. The blood flow of the uteri, their gross appearance and general histology and the density of T lymphocytes were examined on postoperative days 2-28. RESULTS: Macroscopic signs of rejection were apparent from day 5. At the light microscopy level, minimal inflammatory changes were seen from day 5 and massive inflammation was seen from day 10 to day 15. At day 28, necrosis and fibrosis were seen. The density of T lymphocytes (CD3(+)) was increased in the grafted uterus from day 2 in the myometrium and from day 5 in the endometrium. Blood flow in the grafted uteri was reduced from day 15. CONCLUSION: A murine model to study rejection of allogeneic uterus transplants was characterized. Signs of rejection were seen from day 2 to day 5 and severe rejection was seen from day 10 to day 15. The data will be useful in future studies on immunosuppressants in this model. PMID- 16253977 TI - Comparison of serum FSH and Inhibin B levels between adult male dizygotic and monozygotic twins. AB - BACKGROUND: FSH hypersecretion occurs in mothers of dizygotic (DZ) twins. Twinning is inherited via both sexes and transmitted through the female. FSH hypersecretion may thus occur in male DZ twins. METHODS: We assayed FSH and its counter-regulatory hormone, Inhibin B, in 108 adult male DZ and 100 monozygotic (MZ) twins (as controls) and compared our results to published norms. RESULTS: Inhibin B was elevated and higher in DZ compared with MZ twins with similar FSH. CONCLUSION: The normal FSH: Inhibin B endocrine feedback axis is different in adult male DZ twins. This contributes to the theory that the answer to human DZ twinning lies in the actions of FSH and Inhibin, and in their mutual interaction. PMID- 16253978 TI - A comparative randomized trial to assess the impact of oral contraceptive pretreatment on follicular growth and hormone profiles in GnRH antagonist-treated patients. AB - BACKGROUND: This randomized controlled trial was designed to assess the impact of oral contraceptive (OC) scheduling with a GnRH antagonist (ganirelix) regimen on the ovarian response of women undergoing recombinant FSH (rFSH) stimulation for IVF, compared with a non-scheduled ganirelix regimen and a long GnRH agonist (nafarelin) protocol. METHODS: A total of 110 women was treated with an OC and ganirelix, 111 with ganirelix alone and 111 with nafarelin. The OC (containing 30 microg ethinylestradiol/150 microg desogestrel) was taken for 14-28 days and stopped 2 days prior to the start of rFSH treatment. Primary efficiency parameters were the number of cumulus-oocyte complexes (per attempt) and the number of grade 1 or 2 embryos (per attempt). RESULTS: In terms of follicular growth and hormone profiles, the OC-scheduled antagonist regimen mimicked the agonist regimen rather than the (non-scheduled) GnRH antagonist regimen. In the OC-scheduled GnRH antagonist group and the nafarelin group (versus the non scheduled antagonist group), pituitary suppression was more profound at the start of stimulation (P < or = 0.001), there was a slower start of follicular growth (P < or = 0.001), longer stimulation was required (11.7 and 10.3 days respectively versus 9.4; P < or = 0.001), and more rFSH was used (2667 and 2222 IU versus 1966 IU; P < or = 0.001). In the three groups, the number of oocytes was similar (13.1, 12.9 and 11.5 respectively; not significant) as well as the number of good quality embryos (5.1, 5.7 and 5.0 respectively; not significant). CONCLUSION: OC treatment prior to the rFSH/ganirelix regimen can be successfully applied to schedule patients, although more days of stimulation and more rFSH are required than with a non-scheduled GnRH antagonist regimen. PMID- 16253979 TI - Establishing standards for the characterization of human embryonic stem cell lines. AB - As of August 2005, 22 human embryonic stem cell (hESC) lines listed on the National Institutes of Health (NIH) hESC Registry were being distributed to investigators. At a June 2005 meeting of NIH-supported hESC researchers, we proposed that a set of shared standards should be available in order to characterize the cells unambiguously in multiple laboratories. Here, we elaborate such a plan to identify a set of standard methods and to initiate collaborative efforts to validate the standards. The standard assays we propose should be comprehensive enough to ensure that hESC banks can provide a consistent and reliable product for NIH researchers, and inexpensive enough that individual laboratories can afford to use at least some of the methods routinely in their laboratories. We expect that as data accumulate and standards evolve, a core set of tests will become the norm for routine assessment of hESC cultures and that these tests will lay the groundwork for clinical applications of these cells. PMID- 16253980 TI - Cultivation of human embryonic stem cells without the embryoid body step enhances osteogenesis in vitro. AB - Osteogenic cultures of embryonic stem cells (ESCs) are predominately derived from three-dimensional cell spheroids called embryoid bodies (EBs). An alternative method that has been attempted and merits further attention avoids EBs through the immediate separation of ESC colonies into single cells. However, this method has not been well characterized and the effect of omitting the EB step is unknown. Herein, we report that culturing human embryonic stem cells (hESCs) without the EB stage leads to a sevenfold greater number of osteogenic cells and to spontaneous bone nodule formation after 10-12 days. In contrast, when hESCs were differentiated as EBs for 5 days followed by plating of single cells, bone nodules formed after 4 weeks only in the presence of dexamethasone. Furthermore, regardless of the inclusion of EBs, bone matrix formed, including cement line matrix and mineralized collagen, which displayed apatitic mineral (PO4) with calcium-to-phosphorous ratios similar to those of hydroxyapatite and human bone. Together these results demonstrate that culturing hESCs without an EB step can be used to derive large quantities of functional osteogenic cells for bone tissue engineering. PMID- 16253981 TI - Human bone marrow stromal cells express a distinct set of biologically functional chemokine receptors. AB - Stromal cells isolated from bone marrow (BMSCs), often referred to as mesenchymal stem cells, are currently under investigation for a variety of therapeutic applications. However, limited data are available regarding receptors that can influence their homing to and positioning within the bone marrow. In the present study, we found that second passage BMSCs express a unique set of chemokine receptors: three CC chemokine receptors (CCR1, CCR7, and CCR9) and three CXC chemokine receptors (CXCR4, CXCR5, and CXCR6). BMSCs cultured in serum-free medium secrete several chemokine ligands (CCL2, CCL4, CCL5, CCL20, CXCL12, CXCL8, and CX3CL1). The surface-expressed chemokine receptors were functional by several criteria. Stimulation of BMSCs with chemokine ligands triggers phosphorylation of the mitogen-activated protein kinase (e.g., extracellular signal-related kinase [ERK]-1 and ERK-2) and focal adhesion kinase signaling pathways. In addition, CXCL12 selectively activates signal transducer and activator of transcription (STAT)-5 whereas CCL5 activates STAT-1. In cell biologic assays, all of the chemokines tested stimulate chemotaxis of BMSCs, and CXCL12 induces cytoskeleton F-actin polymerization. Studies of culture-expanded BMSCs, for example, 12-16 passages, indicate loss of surface expression of all chemokine receptors and lack of chemotactic response to chemokines. The loss in chemokine receptor expression is accompanied by a decrease in expression of adhesion molecules (ICAM-1, ICAM-2, and vascular cell adhesion molecule 1) and CD157, while expression of CD90 and CD105 is maintained. The change in BMSC phenotype is associated with slowing of cell growth and increased spontaneous apoptosis. These findings suggest that several chemokine axes may operate in BMSC biology and may be important parameters in the validation of cultured BMSCs intended for cell therapy. PMID- 16253982 TI - Olig2 overexpression induces the in vitro differentiation of neural stem cells into mature oligodendrocytes. AB - Differentiation induction of neural stem cells (NSCs) into oligodendrocytes during embryogenesis is the result of a complex interaction between local induction factors and intracellular transcription factors. At the early stage of differentiation, in particular, the helix-loop-helix transcription factors Olig1 and Olig2 have been shown to be essential for oligodendrocyte lineage determination. In view of the possible application of NSCs as a source for remyelinating cell transplants in demyelinating diseases (e.g., multiple sclerosis), in vitro procedures need to be developed to drive the oligodendrocyte differentiation process. Mere culture in medium supplemented with major embryonic oligodendrogenic induction factors, such as Sonic hedgehog, results in oligodendrocyte differentiation of only about 10% of NSCs. We previously showed that induction of Olig1 expression by gene transfection could indeed initiate the first stage of oligodendrocyte differentiation in NSCs, but appeared to be unable to generate fully mature, functional oligodendrocytes. In this study, we transfected NSCs isolated from the embryonic mouse brain with the Olig2 gene and found that the introduced overexpression of Olig2 could induce the development of fully mature oligodendrocytes expressing the transcription factor Nkx2.2 and all major myelin-specific proteins. Moreover, Olig2-transfected NSCs, in contrast to nontransfected NSCs, developed into actively remyelinating oligodendrocytes after transplantation into the corpus callo-sum of long-term cuprizonefed mice, an animal model for demyelination. Our results show that transfection of genes encoding for oligodendrogenic transcription factors can be an efficient way to induce the differentiation of NSCs into functional oligodendrocytes. PMID- 16253983 TI - Neural stem cells express non-neural markers during embryoid body coculture. AB - The capacity of neural stem cells (NSC) to transdifferentiate into a wide range of non-neuronal lineages is the subject of debate. One approach to test NSC plasticity is to ectopically place NSCs in permissive or instructive microenvironments in which the signals driving differentiation of multiple cell types are being elicited. Here we produce embryoid body neurosphere aggregates by combining neurosphere derivatives from fetal mice constitutively expressing green fluorescent protein with embryonic stem (ES) cells isolated from Zin40 mice constitutively expressing nuclear beta-galacosidase. Under these conditions, we assess neurosphere-derivative-immunoreactivity to anti-neurofilament heavy chain, anti-pan-cytokeratin, anti-smooth muscle alpha-actinin and anti-alpha-fetoprotein specific antibodies. Furthermore, we determine lineage-specific transgene expression and undertake fluorescence in situ hybridization to assess ES cell neural stem cell-fusion indices. Our data demonstrate that following coculture in hanging drops with ES cells, neurosphere derivatives display immunoreactivity to non-neural markers, in particular smooth muscle, which is not dependent upon cell cell fusion. These results suggest that given an appropriate environment, NSC may lose their in vivo restrictions and display non-neuronal phenotypes. PMID- 16253984 TI - Hypoxia and serum deprivation-induced apoptosis in mesenchymal stem cells. AB - In recent years, the understanding that regeneration progresses at the level of the myocardium has placed stem cell research at the center stage in cardiology. Despite an increasing interest in cell transplant research, relatively little is known about the biochemical regulation of the stem cell itself after transplantation into an ischemic heart. We demonstrated here, using rat mesenchymal stem cells (MSCs), that cells undergo caspase-dependent apoptosis in response to hypoxia and serum deprivation (SD), which are both components of ischemia in vivo. In particular, the treated cells exhibited mitochondrial dysfunction, including cytochrome C release, loss in DeltaPsim, and Bax accumulation, but in a p53-independent manner. Although the cells treated by hypoxia/SD possess the activity of caspase-8, zIEDT-fmk, a specific caspase-8 inhibitor, failed to inhibit cell apoptosis induced in our system. Taken together, our findings indicate that MSCs are sensitive to hypoxia/SD stimuli that involve changes in mitochondrial integrity and function but are potentially independent of caspase-8. PMID- 16253985 TI - Automated comparative proteomics based on multiplex tandem mass spectrometry and stable isotope labeling. AB - Comparative proteomic approaches using isotopic labeling and MS have become increasingly popular. Conventionally quantification is based on MS or extracted ion chromatogram (XIC) signals of differentially labeled peptides. However, in these MS-based experiments, the accuracy and dynamic range of quantification are limited by the high noise levels of MS/XIC data. Here we report a quantitative strategy based on multiplex (derived from multiple precursor ions) MS/MS data. One set of proteins was metabolically labeled with [13C6]lysine and [15N4]arginine; the other set was unlabeled. For peptide analysis after tryptic digestion of the labeled proteins, a wide precursor window was used to include both the light and heavy versions of each peptide for fragmentation. The multiplex MS/MS data were used for both protein identification and quantification. The use of the wide precursor window increased sensitivity, and the y ion pairs in the multiplex MS/MS spectra from peptides containing labeled and unlabeled lysine or arginine offered more information for, and thus the potential for improving, protein identification. Protein ratios were obtained by comparing intensities of y ions derived from the light and heavy peptides. Our results indicated that this method offers several advantages over the conventional XIC-based approach, including increased sensitivity for protein identification and more accurate quantification with more than a 10-fold increase in dynamic range. In addition, the quantification calculation process was fast, fully automated, and independent of instrument and data type. This method was further validated by quantitative analysis of signaling proteins in the EphB2 pathway in NG108 cells. PMID- 16253986 TI - Differential modification of phosducin protein in degenerating rd1 retina is associated with constitutively active Ca2+/calmodulin kinase II in rod outer segments. AB - Retinitis pigmentosa comprises a heterogeneous group of incurable progressive blinding diseases with unknown pathogenic mechanisms. The retinal degeneration 1 (rd1) mouse is a retinitis pigmentosa model that carries a mutation in a rod photoreceptor-specific phosphodiesterase gene, leading to rapid degeneration of these cells. Elucidation of the molecular differences between rd1 and healthy retinae is crucial for explaining this degeneration and could assist in suggesting novel therapies. Here we used high resolution proteomics to compare the proteomes of the rd1 mouse retina and its congenic, wild-type counterpart at postnatal day 11 when photoreceptor death is profound. Over 3000 protein spots were consistently resolved by two-dimensional gel electrophoresis and subjected to a rigorous filtering procedure involving computer-based spot analyses. Five proteins were accepted as being differentially expressed in the rd1 model and subsequently identified by mass spectrometry. The difference in one such protein, phosducin, related to an altered modification pattern in the rd1 retina rather than to changed expression levels. Additional experiments showed phosducin in healthy retinae to be highly phosphorylated in the dark- but not in the light adapted phase. In contrast, rd1 phosducin was highly phosphorylated irrespective of light status, indicating a dysfunctional rd1 light/dark response. The increased rd1 phosducin phosphorylation coincided with increased activation of calcium/calmodulin-activated protein kinase II, which is known to utilize phosducin as a substrate. Given the increased rod calcium levels present in the rd1 mutation, calcium-evoked overactivation of this kinase may be an early and long sought for step in events leading to photoreceptor degeneration in the rd1 mouse. PMID- 16253987 TI - Transmembrane domain-induced oligomerization is crucial for the functions of syndecan-2 and syndecan-4. AB - The syndecans are known to form homologous oligomers that may be important for their functions. We have therefore determined the role of oligomerization of syndecan-2 and syndecan-4. A series of glutathione S-transferase-syndecan-2 and syndecan-4 chimeric proteins showed that all syndecan constructs containing the transmembrane domain formed SDS-resistant dimers, but not those lacking it. SDS resistant dimer formation was hardly seen in the syndecan chimeras where each transmembrane domain was substituted with that of platelet-derived growth factor receptor (PDGFR). Increased MAPK activity was detected in HEK293T cells transfected with syndecan/PDGFR chimeras in a syndecan transmembrane domain dependent fashion. The chimera-induced MAPK activation was independent of both ligand and extracellular domain, implying that the transmembrane domain is sufficient to induce dimerization/oligomerization in vivo. Furthermore, the syndecan chimeras were defective in syndecan-4-mediated focal adhesion formation and protein kinase Calpha activation or in syndecan-2-mediated cell migration. Taken together, these data suggest that the transmembrane domains are sufficient for inducing dimerization and that transmembrane domain-induced oligomerization is crucial for syndecan-2 and syndecan-4 functions. PMID- 16253988 TI - Crystallographic trapping of the glutamyl-CoA thioester intermediate of family I CoA transferases. AB - Coenzyme A transferases are involved in a broad range of biochemical processes in both prokaryotes and eukaryotes, and exhibit a diverse range of substrate specificities. The YdiF protein from Escherichia coli O157:H7 is an acyl-CoA transferase of unknown physiological function, and belongs to a large sequence family of CoA transferases, present in bacteria to humans, which utilize oxoacids as acceptors. In vitro measurements showed that YdiF displays enzymatic activity with short-chain acyl-CoAs. The crystal structures of YdiF and its complex with CoA, the first co-crystal structure for any Family I CoA transferase, have been determined and refined at 1.9 and 2.0 A resolution, respectively. YdiF is organized into tetramers, with each monomer having an open alpha/beta structure characteristic of Family I CoA transferases. Co-crystallization of YdiF with a variety of CoA thioesters in the absence of acceptor carboxylic acid resulted in trapping a covalent gamma-glutamyl-CoA thioester intermediate. The CoA binds within a well defined pocket at the N- and C-terminal domain interface, but makes contact only with the C-terminal domain. The structure of the YdiF complex provides a basis for understanding the different catalytic steps in the reaction of Family I CoA transferases. PMID- 16253989 TI - Effects of HMGN1 on chromatin structure and SWI/SNF-mediated chromatin remodeling. AB - The dynamic modulation of chromatin structure is determined by many factors, including enzymes that modify the core histone proteins, enzymes that remodel the structure of chromatin, and factors that bind to genomic DNA to affect its structure. Previous work indicates that the nucleosome binding family of high mobility group proteins (HMGN) facilitates the formation of a chromatin structure that is more conducive for transcription. SWI/SNF complexes are ATP-dependent chromatin remodeling enzymes that alter nucleosome structure to facilitate the binding of various regulatory proteins to chromatin. Here we examine the structural consequences of reconstituting chromatin with HMGN1 and the resulting effects on hSWI/SNF function. We demonstrate that HMGN1 decreases the sedimentation velocity of nucleosomal arrays in low ionic strength buffers but has little effect on the structure of more highly folded arrays. We further demonstrate that HMGN1 does not affect SWI/SNF-dependent chromatin remodeling on either mononucleosomes or nucleosomal arrays, indicating that SWI/SNF functions independently of HMGN1. PMID- 16253990 TI - Increased proliferation and altered growth factor dependence of human mammary epithelial cells overexpressing the Gab2 docking protein. AB - The docking protein Gab2 is a proto-oncogene product that is overexpressed in primary breast cancers. To determine the functional consequences of Gab2 overexpression, we utilized the immortalized human mammary epithelial cell line MCF-10A. In monolayer culture, expression of Gab2 at levels comparable with those detected in human breast cancer cells accelerated epidermal growth factor (EGF) induced cell cycle progression and was associated with increased basal Stat5 tyrosine phosphorylation and enhanced and/or more sustained EGF-induced Erk and Akt activation. Three-dimensional Matrigel culture of MCF-10A cells resulted in the formation of polarized, growth-arrested acini with hollow lumina. Under these conditions, Gab2 increased cell proliferation during morphogenesis, leading to significantly larger acini, an effect dependent on Gab2 binding to Grb2 and Shp2 and enhanced by recruitment of the p85 subunit of phosphatidylinositol 3-kinase. Pharmacological inhibition of MEK revealed that, in addition to direct activation of phosphatidylinositol 3-kinase, increased Erk signaling also contributed to Gab2-mediated enhancement of acinar size. In addition, Gab2 overcame the proliferative suppression that normally occurs in late stage cultures and conferred independence of the morphogenetic program from exogenous EGF. Finally, higher levels of Gab2 expression led to the formation of large disorganized structures with defective luminal clearance. These findings support a role for Gab2 in mammary tumorigenesis. PMID- 16253991 TI - Retrograde response to mitochondrial dysfunction is separable from TOR1/2 regulation of retrograde gene expression. AB - Retrograde (RTG) signaling senses mitochondrial dysfunction and initiates readjustments of carbohydrate and nitrogen metabolism through nuclear accumulation of the heterodimeric transcription factors, Rtg1/3p. The RTG pathway is also linked to target of rapamycin (TOR) signaling, among whose activities is transcriptional control of nitrogen catabolite repression (NCR)-sensitive genes. To investigate the connections between these two signaling pathways, we have analyzed rapamycin sensitivity of the expression of the RTG target gene CIT2 and of two NCR-sensitive genes, GLN1 and DAL5, in respiratory-competent (rho+) and incompetent (rho0) yeast cells. Here we have presented evidence that retrograde gene expression is separable from TOR regulation of RTG- and NCR-responsive genes. We showed that expression of these two classes of genes is differentially regulated by glutamate starvation whether in response to mitochondrial dysfunction or induced by rapamycin treatment, as well by glutamine or histidine starvation. We also showed that Lst8p, a component of the TOR1/2 complexes and a negative regulator of the RTG pathway, has multiple roles in the regulation of RTG- and NCR-sensitive genes. Lst8p negatively regulates CIT2 and GLN1 expression, whereas DAL5 expression is independent of Lst8p function. DAL5 expression depends on the GATA transcription factors Gln3p and Gat1p. Gat1p is translocated to the nucleus only upon TOR inhibition by rapamycin. Altogether, these data show that Rtg1/3p, Gln3p, and Gat1p can be differentially regulated through different nutrient-sensing pathways, such as TOR and retrograde signaling, and by multiple factors, such as Lst8p, which is suggested to have a role in connecting the RTG and TOR pathways. PMID- 16253992 TI - The transcription factors AP-1 and Ets are regulators of C3a receptor expression. AB - The anaphylatoxin C3a is a proinflammatory mediator generated during complement activation. The tight control of C3a receptor (C3aR) expression is crucial for the regulation of anaphylatoxin-mediated effects. Key factors regulating constitutive expression of the C3aR in the mast cell line HMC-1 and receptor induction by dibutyryl-cAMP in monomyeloblastic U937 cells were determined by functional characterization of the C3aR promoter. Nucleotides -18 to -285 upstream of the translational start site proved to be critical for promoter activity in HMC-1 cells. Binding sites for the transcription factors AP-1 and Ets could be located. Overexpressed c-Jun/c-Fos (AP-1) and Ets-1 led synergistically to increased promoter activity that was substantially reduced by site-directed mutagenesis of the corresponding elements within the C3aR promoter. In HMC-1 cells, Ets interacted directly with the predicted binding motif of the C3aR promoter as determined by electromobility shift assays. AP-1 binding to the C3aR promoter was augmented during C3aR induction in U937 cells. A retroviral gene transfer system was used to express a dominant negative mutant of Ets-1 in these cells. The resulting cells failed to up-regulate the C3aR after stimulation with dibutyryl-cAMP and showed decreased AP-1 binding, suggesting that Ets acts here indirectly. Thus, it was established that Ets and the AP-1 element mediates dibutyryl-cAMP induction of C3aR promoter activity, hence providing a mechanistic explanation of dibutyryl-cAMP-dependent up-regulation of C3aR expression. In conclusion, this study demonstrates an important role of AP-1 and a member of the Ets family in the transcriptional regulation of C3aR expression, a prerequisite for the ability of C3a to participate in immunomodulation and inflammation. PMID- 16253993 TI - The structure of Prp40 FF1 domain and its interaction with the crn-TPR1 motif of Clf1 gives a new insight into the binding mode of FF domains. AB - The yeast splicing factor Prp40 (pre-mRNA processing protein 40) consists of a pair of WW domains followed by several FF domains. The region comprising the FF domains has been shown to associate with the 5' end of U1 small nuclear RNA and to interact directly with two proteins, the Clf1 (Crooked neck-like factor 1) and the phosphorylated repeats of the C-terminal domain of RNA polymerase II (CTD RNAPII). In this work we reported the solution structure of the first FF domain of Prp40 and the identification of a novel ligand-binding site in FF domains. By using chemical shift assays, we found a binding site for the N-terminal crooked neck tetratricopeptide repeat of Clf1 that is distinct and structurally separate from the previously identified CTD-RNAPII binding pocket of the FBP11 (formin binding protein 11) FF1 domain. No interaction, however, was observed between the Prp40 FF1 domain and three different peptides derived from the CTD-RNAPII protein. Indeed, the equivalent CTD-RNAPII-binding site in the Prp40 FF1 domain is predominantly negatively charged and thus unfavorable for an interaction with phosphorylated peptide sequences. Sequence alignments and phylogenetic tree reconstructions using the FF domains of three functionally related proteins, Prp40, FBP11, and CA150, revealed that Prp40 and FBP11 are not orthologous proteins and supported the different ligand specificities shown by their respective FF1 domains. Our results also revealed that not all FF domains in Prp40 are functionally equivalent. We proposed that at least two different interaction surfaces exist in FF domains that have evolved to recognize distinct binding motifs. PMID- 16253994 TI - Crystal structure of the human B-form low molecular weight phosphotyrosyl phosphatase at 1.6-A resolution. AB - The crystal structure of HPTP-B, a human isoenzyme of the low molecular weight phosphotyrosyl phosphatase (LMW PTPase) is reported here at a resolution of 1.6 A. This high resolution structure of the second human LMW PTPase isoenzyme provides the opportunity to examine the structural basis of different substrate and inhibitor/activator responses. The crystal packing of HPTP-B positions a normally surface-exposed arginine in a position equivalent to the tyrosyl substrate. A comparison of all deposited crystallographic coordinates of these PTPases reveals three atomic positions within the active site cavity occupied by hydrogen bond donor or acceptor atoms on bound molecules, suggesting useful design elements for synthetic inhibitors. A selection of inhibitor and activator molecules as well as small molecule and peptide substrates were tested against each human isoenzyme. These results along with the crystal packing seen in HPTP-B suggest relevant sequence elements in the currently unknown target sequence. PMID- 16253995 TI - Defective tumor necrosis factor-alpha-dependent control of astrocyte glutamate release in a transgenic mouse model of Alzheimer disease. AB - The cytokine tumor necrosis factor-alpha (TNFalpha) induces Ca2+-dependent glutamate release from astrocytes via the downstream action of prostaglandin (PG) E2. By this process, astrocytes may participate in intercellular communication and neuromodulation. Acute inflammation in vitro, induced by adding reactive microglia to astrocyte cultures, enhances TNFalpha production and amplifies glutamate release, switching the pathway into a neurodamaging cascade (Bezzi, P., Domercq, M., Brambilla, L., Galli, R., Schols, D., De Clercq, E., Vescovi, A., Bagetta, G., Kollias, G., Meldolesi, J., and Volterra, A. (2001) Nat. Neurosci. 4, 702-710). Because glial inflammation is a component of Alzheimer disease (AD) and TNFalpha is overexpressed in AD brains, we investigated possible alterations of the cytokine-dependent pathway in PDAPP mice, a transgenic model of AD. Glutamate release was measured in acute hippocampal and cerebellar slices from mice at early (4-month-old) and late (12-month-old) disease stages in comparison with age-matched controls. Surprisingly, TNFalpha-evoked glutamate release, normal in 4-month-old PDAPP mice, was dramatically reduced in the hippocampus of 12-month-old animals. This defect correlated with the presence of numerous beta amyloid deposits and hypertrophic astrocytes. In contrast, release was normal in cerebellum, a region devoid of beta-amyloid deposition and astrocytosis. The Ca2+ dependent process by which TNFalpha evokes glutamate release in acute slices is distinct from synaptic release and displays properties identical to those observed in cultured astrocytes, notably PG dependence. However, prostaglandin E2 induced normal glutamate release responses in 12-month-old PDAPP mice, suggesting that the pathology-associated defect involves the TNFalpha-dependent control of secretion rather than the secretory process itself. Reduced expression of DENN/MADD, a mediator of TNFalpha-PG coupling, might account for the defect. Alteration of this neuromodulatory astrocytic pathway is described here for the first time in relation to Alzheimer disease. PMID- 16253996 TI - Cdc42 induces activation loop phosphorylation and membrane targeting of mixed lineage kinase 3. AB - Mixed lineage kinase 3 (MLK3) functions as a mitogen-activated protein kinase kinase kinase to activate multiple mitogen-activated protein kinase pathways. Our current studies demonstrate that lack of MLK3 blocks signaling of activated Cdc42 to c-Jun N-terminal kinase, giving strong support for the idea that Cdc42 is a physiological activator of MLK3. We show herein that Cdc42, in a prenylation dependent manner, targets MLK3 from a perinuclear region to membranes, including the plasma membrane. Cdc42-induced membrane targeting of MLK3 is independent of MLK3 catalytic activity but depends upon an intact Cdc42/Rac-interactive binding motif, consistent with MLK3 membrane translocation being mediated through direct binding of Cdc42. Phosphorylation of the activation loop of MLK3 requires MLK3 catalytic activity and is induced by Cdc42 in a prenylation-independent manner, arguing that Cdc42 binding is sufficient for activation loop autophosphorylation of MLK3. However, membrane targeting is necessary for full activation of MLK3 and maximal signaling to JNK. We previously reported that MLK3 is autoinhibited through an interaction between its N-terminal SH3 domain and a proline-containing sequence found between the leucine zipper and the CRIB motif of MLK3. Thus we propose a model in which GTP-bound Cdc42/Rac binds MLK3 and disrupts SH3-mediated autoinhibition leading to dimerization and activation loop autophosphorylation. Targeting of this partially active MLK3 to membranes likely results in additional phosphorylation events that fully activate MLK3 and its ability to maximally signal through the JNK pathway. PMID- 16253997 TI - CpG-binding protein (CXXC finger protein 1) is a component of the mammalian Set1 histone H3-Lys4 methyltransferase complex, the analogue of the yeast Set1/COMPASS complex. AB - CpG-binding protein (CXXC finger protein 1 (CFP1)) binds to DNA containing unmethylated CpG motifs and is required for mammalian embryogenesis, normal cytosine methylation, and cellular differentiation. Studies were performed to identify proteins that interact with CFP1 to gain insight into the molecular function of this protein. Immunoprecipitation and mass spectrometry reveal that human CFP1 associates with a approximately 450-kDa complex that contains the mammalian homologues of six of the seven components of the Set1/COMPASS complex, the sole histone H3-Lys4 methyltransferase in yeast. In vitro assays demonstrate that the human Set1/CFP1 complex is a histone methyltransferase that produces mono-, di-, and trimethylated histone H3 at Lys4. Confocal microscopy reveals that CFP1 and Set1 co-localize to nuclear speckles associated with euchromatin. A Set1 complex of reduced mass persists in murine embryonic stem cells lacking CFP1. These cells carry elevated levels of methylated histone H3-Lys4 and reduced levels of methylated histone H3-Lys9. Together with the previous finding of reduced levels of cytosine methylation, these data indicate that cells lacking CFP1 contain reduced levels of heterochromatin. Furthermore, ES cells lacking CFP1 exhibit a 4-fold excess of histone H3-Lys4 methylation following induction of differentiation, indicating that CFP1 restricts the activity of the Set1 histone methyltransferase complex. These results reveal a mammalian counterpart to the yeast Set1/COMPASS complex. The presence of CFP1 in this complex implicates this protein as a critical epigenetic regulator of histone modification in addition to cytosine methylation and reveals one mechanism by which this protein intersects with the epigenetic machinery. PMID- 16253998 TI - Prototypical type I E-cadherin and type II cadherin-7 mediate very distinct adhesiveness through their extracellular domains. AB - Using a dual pipette assay that measures the force required to separate adherent cell doublets, we have quantitatively compared intercellular adhesiveness mediated by Type I (E- or N-cadherin) or Type II (cadherin-7 or -11) cadherins. At similar cadherin expression levels, cells expressing Type I cadherins adhered much more rapidly and strongly than cells expressing Type II cadherins. Using chimeric cadherins, we found that the extracellular domain exerts by far the dominant effect on cell adhesivity, that of E-cadherin conferring high adhesivity, and that of cadherin-7 conferring low adhesivity. Type I cadherins were incorporated to a greater extent into detergent-insoluble cytoskeletal complexes, and their cytoplasmic tails were much more effective in disrupting strong adherent junctions, suggesting that Type II cadherins form less stable complexes with beta-catenin. The present study demonstrates compellingly, for the first time, that cadherins are dramatically different in their ability to promote intercellular adhesiveness, a finding that has profound implications for the regulation of tissue morphogenesis. PMID- 16253999 TI - Interaction of SPIN90 with the Arp2/3 complex mediates lamellipodia and actin comet tail formation. AB - The appropriate regulation of the actin cytoskeleton is essential for cell movement, changes in cell shape, and formation of membrane protrusions like lamellipodia and filopodia. Moreover, several regulatory proteins affecting actin dynamics have been identified in the motile regions of cells. Here, we provide evidence for the involvement of SPIN90 in the regulation of actin cytoskeleton and actin comet tail formation. SPIN90 was distributed throughout the cytoplasm in COS-7 cells, but exposing the cells to platelet-derived growth factor (PDGF) caused a redistribution of SPIN90 to the cell cortex and the formation of lamellipodia (or membrane ruffles), both of which were dramatically inhibited in SPIN90-knockdown cells. In addition, the binding of the C terminus of SPIN90 with both the Arp2/3 complex (actin-related proteins Arp 2 and Arp 3) and G-actin activates the former, leading to actin polymerization in vitro. And when coexpressed with phosphatidylinositol 4-phosphate 5 kinase, SPIN90 was observed within actin comet tails. Taken these findings suggest that SPIN90 participates in reorganization of the actin cytoskeleton and in actin-based cell motility. PMID- 16254000 TI - Biochemical and motile properties of Myo1b splice isoforms. AB - Myo1b is a widely expressed myosin-I isoform that concentrates on endosomal and ruffling membranes and is thought to play roles in membrane trafficking and dynamics. Myo1b is alternatively spliced within the regulatory domain of the molecule, yielding isoforms with six (myo1b(a)), five (myo1b(b)), or four (myo1b(c)) non-identical IQ motifs. The calmodulin binding properties of the myo1b IQ motifs have not been investigated, and the mechanical and cell biological consequences of alternative splicing are not known. Therefore, we expressed the alternatively spliced myo1b isoforms truncated after the final IQ motif and included a sequence at their C termini that is a substrate for bacterial biotin ligase. Site-specific biotinylation allows us to specifically attach the myosin to motility surfaces via a biotin-streptavidin linkage. We measured the ATPase and motile properties of the recombinant myo1b splice isoforms, and we correlated these properties with calmodulin binding. We confirmed that calcium-dependent changes in the ATPase activity are due to calcium binding to the calmodulin closest to the motor. We found that calmodulin binds tightly to some of the IQ motifs (Kd < 0.2 microM) and very weakly to the others (Kd > 5 microM), suggesting that a subset of the IQ motifs are not calmodulin bound under physiological conditions. Finally, we found the in vitro motility rate to be dependent on the myo1b isoform and the calmodulin concentration and that the myo1b regulatory domain acts as a rigid lever arm upon calmodulin binding to the high affinity and low affinity IQ motifs. PMID- 16254001 TI - GnRH agonist for triggering final oocyte maturation in the GnRH antagonist ovarian hyperstimulation protocol: a systematic review and meta-analysis. AB - Triggering final oocyte maturation with GnRH agonist during ovarian stimulation is feasible when inhibition of premature LH surge is performed with GnRH antagonists, and we aimed to systematically collate evidence on the clinical efficacy of GnRH agonist triggering in patients undergoing assisted reproduction in GnRH antagonist protocols. Twenty-three publications were identified by a comprehensive literature search that included PubMed, Embase and the Cochrane Library. Three publications out of 23 fulfilled the inclusion criteria for meta analysis, which were (i) prospective, randomized controlled study design; (ii) stimulation with gonadotropins for induction of multifollicular development; (iii) suppression of endogenous LH by a GnRH antagonist; (iv) triggering of final oocyte maturation with GnRH agonist; (v) control group randomized to receive HCG for final oocyte maturation and (vi) any means of luteal phase support other than HCG. The participants were normoovulatory women undergoing IVF. The outcomes assessed were clinical pregnancy per randomized patient; number of oocytes retrieved; proportion of metaphase II oocytes; fertilization rate; embryo quality score; first trimester abortion rate; ovarian hyperstimulation syndrome (OHSS) incidence. Results are presented as combined standardized differences of the mean and combined odds ratios, as appropriate, with 95% confidence intervals. No significant difference was found for the number of oocytes retrieved (-0.94, 0.33-0.14), proportion of metaphase II oocytes (-0.03, -0.58-0.52), fertilization rate (0.15, -0.09-0.38) or embryo quality score (0.05, -0.18-0.29). No OHSS occurred in two of the studies, whereas in one study OHSS incidence was not reported. Thus from the available data, no conclusion can be drawn as regards OHSS incidence after GnRH agonist triggering. In comparison to HCG, GnRH agonist administration is associated with a significantly reduced likelihood of achieving a clinical pregnancy (0.21, 0.05-0.84; P = 0.03). The odds of first trimester pregnancy loss is increased after GnRH agonist triggering; however, the confidence interval crosses unity (11.51, 0.95-138.98; P = 0.05). In conclusion, the use of GnRH agonist to trigger final oocyte maturation in IVF, where inhibition of premature LH surge is achieved with GnRH antagonists, yields a number of oocytes capable to undergo fertilization and subsequent embryonic cleavage, which is comparable to that achieved with HCG. However, the likelihood of an ongoing clinical pregnancy after GnRH agonist triggering is significantly lower as compared to standard HCG treatment. PMID- 16254002 TI - Consequences of mutations in the non-coding RMRP RNA in cartilage-hair hypoplasia. AB - Cartilage-hair hypoplasia (CHH), also known as metaphyseal chondrodysplasia McKusick type (OMIM no. 250250), is an autosomal recessive, multi-systemic disease characterized by disproportionate short stature, fine and sparse hair, deficient cellular immunity and a predisposition to malignancy. It is caused by mutations in RMRP, the RNA component of the ribonucleoprotein complex RNase MRP, and, thus, CHH represents one of few Mendelian disorders caused by mutations in a nuclear encoded, non-coding RNA. While studies in yeast indicate that RMRP contributes to diverse cellular functions, the pathogenesis of the human condition is unknown. Studies of our CHH patient cohort revealed mutations in both the promoter and the transcribed region of RMRP. While mutations in the promoter abolished transcription in vitro, RMRP RNA levels in patients with transcribed mutations were also decreased suggesting an unstable RNA. RMRP mutations introduced into the yeast ortholog, NME1, exhibited normal mitochondrial function, chromosomal segregation and cell cycle progression, while a CHH fibroblast cell line exhibited normal mitochondrial content. However, the most commonly found mutation in CHH patients, 70A>G, caused an alteration in ribosomal processing by altering the ratio of the short versus the long form of the 5.8S rRNA in yeast. Transcriptional profiling of CHH patient RNAs showed upregulation of several cytokines and cell cycle regulatory genes, one of which has been implicated in chondrocyte hypertrophy. These data suggest that alteration of ribosomal processing in CHH is associated with altered cytokine signalling and cell cycle progression in terminally differentiating cells in the lymphocytic and chondrocytic cell lineages. PMID- 16254003 TI - Discovery of 2,5-dimethoxy-substituted 5-bromopyridyl thiourea (PHI-236) as a potent broad-spectrum anti-human immunodeficiency virus microbicide. AB - The increased risk of heterosexual transmission of human immunodeficiency virus-1 (HIV-1) has prompted the search for safe and effective female-controlled vaginal microbicides. Because endogenous reverse transcription is implicated in augmenting the sexual transmission of HIV-1, potential microbicides should have the inherent ability to optimally inhibit both wild-type and drug-resistant mutant strains of HIV-1. N-[2-(2,5-dimethoxyphenylethyl)]-N'-[2-(5-bromopyridyl)] thiourea (PHI-236) is a rationally designed non-nucleoside inhibitor of HIV-1 reverse transcriptase (NNRTI) that was deduced from changes in binding pocket size, shape and residue character that result from clinically observed NNRTI resistance mutations. PHI-236 displayed high-binding affinity (Ludi K(i) = 0.07 microM) for HIV-1 RT and robust anti-HIV activity against the wild type (IC50 = <0.001 microM) as well as primary clinical isolates (IC50 = 0.009-0.04 microM) carrying multiple RT gene mutations associated with NRTI and NNRTI resistance. PHI-236 displayed high-selectivity index against human vaginal and cervical epithelial cells and did not affect human sperm functions. In the humanized severe combined immunodeficient mouse model for HIV/acquired immune deficiency syndrome (AIDS), pretreatment of HIV-1 (BaL)-infected human monocytes and semen with PHI-236 prevented the systemic infection via the vaginal route. PHI-236 has particular clinical utility as a non-spermicidal microbicide as well as a prophylactic antiviral agent to inactivate cell-free and cell-associated HIV-1 in semen before assisted reproductive technology procedures. PMID- 16254004 TI - Universal DNA primers amplify bacterial DNA from human fetal membranes and link Fusobacterium nucleatum with prolonged preterm membrane rupture. AB - A large number of bacterial species have been identified in fetal membranes after preterm labour (PTL) associated with intrauterine infection by microbiological culture. In this study, we have investigated a molecular and bioinformatic approach to organism identification which surmounts the need for specific and diverse microbiological culture conditions required by conventional methods. Samples of fetal membranes were taken from 37 preterm infants, and 6 normal term controls delivered by caesarean section, in which bacteria had been detected by in situ hybridization of 16S ribosomal RNA using a generic probe. Degenerate primers were designed to amplify bacterial 16S ribosomal DNA by PCR and used to amplify bacterial DNA from human fetal membranes. Amplicons were cloned, sequenced and bacteria were identified bioinformatically by comparison of sequences with known bacterial DNA genomes. In situ hybridization using an organism specific probe was then used to confirm the presence of the commonest identified organism in tissue samples. Bacterial DNA amplified from 15/43 samples, all from preterm deliveries, and the bioinformatic approach identified organisms in all cases. Multiple bacteria were identified including Mycoplasma hominis, Pasturella multocida, Pseudomonas PH1, Escherichia coli and Prevotella bivia. The commonest organism Fusobacterium nucleatum was found in 9/15 (60%) of samples. Ten of the 12 samples obtained after prolonged membrane rupture were positive for bacterial DNA, and 7 of these (70%) contained DNA from F. nucleatum. Bacteria from fetal membranes may be identified by molecular and bioinformatic methods. Further work is warranted to investigate the apparent linkage between F. nucleatum, fetal membrane rupture and preterm delivery. PMID- 16254005 TI - Differential regulation of interleukins IL-13 and IL-15 by ovarian steroids, TNF alpha and TGF-beta in human endometrial epithelial and stromal cells. AB - Based on the endometrial spatial and temporal expression of interleukins (ILs) IL 13 and IL-15 during the normal menstrual cycle, we hypothesized that ovarian steroids and non-steroidal factors regulate their expression in a cell-specific manner. To test this hypothesis and determine IL-13/IL-15 actions, we used endometrial epithelial (EEC) and stromal (ESC) cells isolated and cultured under defined conditions. We confirmed the expression of IL-13 and IL-15 in these cells and further demonstrated that 17beta estradiol (E2), medroxyprogesterone acetate (MPA) and their combination differentially regulated their mRNA expression and protein production in a time- and cell-specific manner (P < 0.05). We also showed that tumour necrosis factor-alpha (TNF-alpha; 10 and 25 ng/ml) and transforming growth factor-beta (TGF-beta; 1 and 5 ng/ml), cytokines with inflammatory and immune regulatory functions in a cell- and dose-dependent manner regulate the expression of IL-13 and IL-15 (P < 0.05). Functionally, IL-13 and IL-15 1-100 ng/ml displayed a limited mitogenic activity towards EEC and ESC; however, they regulated the expression of TNF receptor type 1 (TNFR) mRNA and soluble protein in a cell-specific manner (P < 0.05). We conclude that ovarian steroids, TNF alpha and TGF-beta act as key regulators of endometrial IL-13 and IL-15 expression which act locally regulating TNFR expression in a cell-specific manner. Based on these findings, we conclude that IL-13/IL-15, either alone or through their interactions with other cytokines, influence the outcome of endometrial inflammatory/immune responses during the normal menstrual cycle, and due to their altered expression may extend these processes in dysfunctional bleeding and endometriosis. PMID- 16254006 TI - Post-training quality of syndromic management of sexually transmitted infections by chemists and druggists in Pokhara, Nepal: is it satisfactory? AB - OBJECTIVE: Using simulated client and provider interview methods, this study assessed chemists and druggists' post-training management quality of syndromic sexually transmitted infections focusing on the areas of privacy maintaining, encouraging, history taking, counseling, referral practice, partner notification, and drug prescribing and then compared the findings of two methods. DESIGN: Forty five pharmacies from a list of 75 in Pokhara, who collected sexually transmitted infections data during 1999, were selected randomly. First simulated client successfully presented either urethral or vaginal discharge syndrome at 37 pharmacies and recorded the events of whole encounter into an observation form within 20 minutes. Later 39 chemists and druggists were interviewed by a pre tested semi-structured questionnaire. MAIN MEASURE: Results were reported mainly by numbers and corresponding percentages. For comparative purpose, P values were also shown. RESULTS: Overall, interview method revealed satisfactory knowledge of chemists and druggists for management of sexually transmitted infections except drug prescribing but their actual behaviors, revealed by simulated client method, indicated lower quality and differed significantly in the areas of encouraging, history taking, counseling, referral practice, and partner notification. Both methods indicated very poor qualities of drug prescribing. CONCLUSION: Retained knowledge of chemists and druggists for syndromic management of sexually transmitted infections were not applied to simulated client in actual practice. They should not prescribe drugs for patients of sexually transmitted infections, except referring to the doctors/hospitals. Continuous monitoring and further motivations for them may improve syndromic management quality of sexually transmitted infections. Moreover, depending on the purpose of study, various methods should be applied simultaneously to reach a better conclusion. PMID- 16254007 TI - Patent foramen ovale closure in patients with cryptogenic thrombo-embolic events using the Cardia PFO occluder. AB - AIMS: We report our experience with three generations of the Cardia patent foramen ovale (PFO) occluder in patients with cryptogenic thrombo-embolic events (TE). METHODS AND RESULTS: Between 1998 and 2004, interventional PFO closure was attempted in 403 patients. Prior to PFO closure, 605 TE occurred, translating into an annual incidence of 3.1%. PFO closure was successful in all patients. Peri-procedural complications occurred in eight patients (2.0%). At 6 months follow-up, residual shunt was present in 10.8% of patients. Transient thrombi developed on 10 devices (predominantly generation II) and asymptomatic wire fractures were detected in 14 cases (generation I and II). The annual incidence of recurrent TE was 2.0% (n=13). Atrial septal aneurysm and prior device-related thrombus formation were identified as predictors of recurrent TE. CONCLUSION: Owing to technical improvements and modified adjunctive pharmacotherapy, the rate of thrombus formation has declined and wire fractures are virtually absent in generation III devices. The overall rate of recurrent TE is reduced by transcatheter PFO closure with the Cardia PFO occluder, and seems comparable to recurrence rates reported for other devices used for this purpose. PMID- 16254008 TI - Do muscle blood flow detectors link breathing to oxygen consumption in exercise? PMID- 16254009 TI - Catecholamine neurones in rats modulate sleep, breathing, central chemoreception and breathing variability. AB - Brainstem catecholamine (CA) neurones have wide projections and an arousal-state dependent activity pattern. They are thought to modulate the processing of sensory information and also participate in the control of breathing. Mice with lethal genetic defects that include CA neurones have abnormal respiratory control at birth. Also the A6 region (locus coeruleus), which contains CA neurones sensitive to CO(2) in vitro, is one of many putative central chemoreceptor sites. We studied the role of CA neurones in the control of breathing during sleep and wakefulness by specifically lesioning them with antidopamine beta-hydroxylase saporin (DBH-SAP) injected via the 4th ventricle. After 3 weeks there was a 73 84% loss of A5, A6 and A7 tyrosine hydroxylase (TH) immunoreactive (ir) neurones along with 56-60% loss of C1 and C2 phenyl ethanolamine-N-methyltransferase (PNMT)-ir neurones. Over the 3 weeks, breathing frequency decreased significantly during air and 3 or 7% CO(2) breathing in both wakefulness and non-REM (NREM) sleep. The rats spent significantly less time awake and more time in NREM sleep. REM sleep time was unaffected. The ventilatory response to 7% CO(2) was reduced significantly in wakefulness at 7, 14 and 21 days (-28%) and in NREM sleep at 14 and 21 days (-26%). Breathing variability increased in REM sleep but not in wakefulness or NREM sleep. We conclude that CA neurones (1) promote wakefulness, (2) participate in central respiratory chemoreception, (3) stimulate breathing frequency, and (4) minimize breathing variability in REM sleep. PMID- 16254010 TI - Hypoxic pulmonary vasoconstriction: mechanisms and controversies. AB - The pulmonary circulation differs from the systemic in several important aspects, the most important being that pulmonary arteries constrict to moderate physiological (20-60 mmHg PO2) hypoxia, whereas systemic arteries vasodilate. This phenomenon is called hypoxic pulmonary vasoconstriction (HPV), and is responsible for maintaining the ventilation-perfusion ratio during localized alveolar hypoxia. In disease, however, global hypoxia results in a detrimental increase in total pulmonary vascular resistance, and increased load on the right heart. Despite many years of study, the precise mechanisms underlying HPV remain unresolved. However, as we argue below, there is now overwhelming evidence that hypoxia can stimulate several pathways leading to a rise in the intracellular Ca2+ concentration ([Ca2+]i) in pulmonary artery smooth muscle cells (PASMC). This rise in [Ca2+]i is consistently found to be relatively small, and HPV seems also to require rho kinase-mediated Ca2+ sensitization. There is good evidence that HPV also has an as yet unexplained endothelium dependency. In this brief review, we highlight selected recent findings and ongoing controversies which continue to animate the study of this remarkable and unique response of the pulmonary vasculature to hypoxia. PMID- 16254011 TI - Reduced mitochondrial coupling in vivo alters cellular energetics in aged mouse skeletal muscle. AB - The mitochondrial theory of ageing proposes that the accumulation of oxidative damage to mitochondria leads to mitochondrial dysfunction and tissue degeneration with age. However, no consensus has emerged regarding the effects of ageing on mitochondrial function, particularly for mitochondrial coupling (P/O). One of the main barriers to a better understanding of the effects of ageing on coupling has been the lack of in vivo approaches to measure P/O. We use optical and magnetic resonance spectroscopy to independently quantify mitochondrial ATP synthesis and O2 uptake to determine in vivo P/O. Resting ATP demand (equal to ATP synthesis) was lower in the skeletal muscle of 30-month-old C57Bl/6 mice compared to 7-month old controls (21.9 +/- 1.5 versus 13.6 +/- 1.7 nmol ATP (g tissue)(-1) s(-1), P = 0.01). In contrast, there was no difference in the resting rates of O2 uptake between the groups (5.4 +/- 0.6 versus 8.4 +/- 1.6 nmol O2 (g tissue)(-1) s(-1)). These results indicate a nearly 50% reduction in the mitochondrial P/O in the aged animals (2.05 +/- 0.07 versus 1.05 +/- 0.36, P = 0.02). The higher resting ADP (30.8 +/- 6.8 versus 58.0 +/- 9.5 micromol g(-1), P = 0.05) and decreased energy charge (ATP/ADP) (274 +/- 70 versus 84 +/- 16, P = 0.03) in the aged mice is consistent with an impairment of oxidative ATP synthesis. Despite the reduced P/O, uncoupling protein 3 protein levels were not different in the muscles of the two groups. These results demonstrate reduced mitochondrial coupling in aged skeletal muscle that alters cellular metabolism and energetics. PMID- 16254012 TI - Long-QT syndrome-related sodium channel mutations probed by the dynamic action potential clamp technique. AB - Long-QT3 syndrome (LQT3) is linked to cardiac sodium channel gene (SCN5A) mutations. In this study, we used the 'dynamic action potential clamp' (dAPC) technique to effectively replace the native sodium current (I(Na)) of the Priebe Beuckelmann human ventricular cell model with wild-type (WT) or mutant I(Na) generated in a human embryonic kidney (HEK)-293 cell that is voltage clamped by the free-running action potential of the ventricular cell. We recorded I(Na) from HEK cells expressing either WT or LQT3-associated Y1795C or A1330P SCN5A at 35 degrees C, and let this current generate and shape the action potential (AP) of subepicardial, mid-myocardial and subendocardial model cells. The HEK cell's endogenous background current was completely removed by a real-time digital subtraction procedure. With WT I(Na), AP duration (APD) was longer than with the original Priebe-Beuckelmann model I(Na), due to a late I(Na) component of approximately 30 pA that could not be revealed with conventional voltage-clamp protocols. With mutant I(Na), this late component was larger ( approximately 100 pA), producing a marked increase in APD ( approximately 70-80 ms at 1 Hz for the subepicardial model cell). The late I(Na) magnitude showed reverse frequency dependence, resulting in a significantly steeper APD-frequency relation in the mutant case. AP prolongation was more pronounced for the mid-myocardial cell type, resulting in increased APD dispersion for each of the mutants. For both mutants, a 2 s pause following rapid (2 Hz) pacing resulted in distorted AP morphology and beat-to-beat fluctuations of I(Na). Our dAPC data directly demonstrate the arrhythmogenic nature of LQT3-associated SCN5A mutations. PMID- 16254013 TI - Acute response to intracisternal bupivacaine in patients with refractory pain of the head and neck. AB - Continuous intracisternal infusion of bupivacaine for the management of intractable pain of the head and neck is effective in controlling pain in this patient group. With the catheter tip being located at the height of the C1 vertebral body, autonomic regulatory information may also be influenced by the infusion of bupivacaine. By combining direct sampling of cerebrospinal fluid (CSF), via a percutaneously placed catheter in the cisterna magna, with a noradrenaline and adrenaline isotope dilution method for examining sympathetic and adrenal medullary activity, we were able to quantify the release of brain neurotransmitters and examine efferent sympathetic nervous outflow in patients following intracisternal administration of bupivacaine. Despite severe pain, sympathetic and adrenal medullary activities were well within normal range (4.2 +/- 0.6 and 0.7 +/- 0.2 nmol min(-1), respectively, mean +/-S.E.M.). Intracisternal bupivacaine administration caused an almost instantaneous elevation in mean arterial blood pressure, increasing by 17 +/- 7 mmHg after 10 min (P < 0.01). Heart rate increased in parallel (17 +/- 5 beats min(-1)), and these changes coincided with an increase in sympathetic nervous activity, peaking with an approximately 50% increase over resting level 10 min after injection (P < 0.01). CSF levels of GABA were reduced following bupivacaine (P < 0.05). CSF catecholamines and serotonin, and EEG, remained unaffected. These results show that acutely administered bupivacaine in the cisterna magna of chronic pain sufferers leads to an activation of the sympathetic nervous system. The results suggest that the haemodynamic consequences occur as a result of interference with the neuronal circuitry in the brainstem. Although these effects are transient, they warrant caution at the induction of intracisternal local anaesthesia. PMID- 16254014 TI - Multifunction steroid receptor coactivator, E6-associated protein, is involved in development of the prostate gland. AB - In this study we report that deletion of E6-associated protein (E6-AP) in mice results in a smaller prostate gland compared with that in normal wild-type animals. To investigate the mechanism(s) by which E6-AP affects prostate gland growth and development, we carried out both in vitro and in vivo experiments. In this study we show that E6-AP interacts with androgen receptor (AR) in a hormone dependent manner and enhances the transactivation function of AR. Our in vivo data from E6-AP-null prostate glands show that the level of AR protein is elevated while the level of the AR target protein, probasin, is decreased. In contrast, the level of AR protein is decreased, and its target protein is increased in an E6-AP-overexpressing stable cell line, suggesting that E6-AP modulates both the protein level and the activity of AR. In addition, we show that the levels of phosphatidylinositol 3-kinase, total Akt, and phosphorylated Akt are decreased in E6-AP-null prostate, suggesting that E6-AP deletion down regulates the signaling of the phosphatidylinositol 3-kinase-Akt pathway. We also show that RhoA negatively regulates AR function, and RhoA levels are increased in E6-AP-null prostate. Furthermore, expression levels of p53, Bax, active caspases, and apoptotic index are increased in E6-AP-null prostate. Collectively, our data suggest that E6-AP deletion attenuates the growth and development of the prostate gland by interfering with AR function as well as by stimulating p53-mediated apoptosis. PMID- 16254015 TI - Thyroid hormone-regulated target genes have distinct patterns of coactivator recruitment and histone acetylation. AB - Thyroid hormone receptors (TRs) are ligand-regulated transcription factors that bind to thyroid hormone response elements of target genes. Upon ligand binding, they recruit coactivator complexes that increase histone acetylation and recruit RNA polymerase II (Pol II) to activate transcription. Recent studies suggest that nuclear receptors and coactivators may have temporal recruitment patterns on hormone response elements, yet little is known about the nature of the patterns at multiple endogenous target genes. We thus performed chromatin immunoprecipitation assays to investigate coactivator recruitment and histone acetylation patterns on the thyroid hormone response elements of four endogenous target genes (GH, sarcoplasmic endoplasmic reticulum calcium-adenosine triphosphatase, phosphoenolpyruvate carboxykinase, and cholesterol 7alpha hydroxylase) in a rat pituitary cell line that expresses TRs. We found that TRbeta, several associated coactivators (steroid receptor coactivator-1, glucocorticoid receptor interacting protein-1, and TR-associated protein 220), and RNA Pol II were rapidly recruited to thyroid hormone response elements as early as 15 min after T3 addition. When the four target genes were compared, we observed differences in the types and temporal patterns of recruited coactivators and histone acetylation. Interestingly, the temporal pattern of RNA Pol II was similar for three genes studied. Our findings suggest that thyroid hormone regulated target genes may have distinct patterns of coactivator recruitment and histone acetylation that may enable highly specific regulation. PMID- 16254018 TI - The clinical heterogeneity of immune sensory and autonomic neuropathies with (or without) sicca. PMID- 16254019 TI - The leaf size-twig size spectrum of temperate woody species along an altitudinal gradient: an invariant allometric scaling relationship. AB - BACKGROUND AND AIMS: The leaf size-twig size spectrum is one of the leading dimensions of plant ecological variation, and now it is under development. The purpose of this study was to test whether the relationship between leaf size and twig size is isometric or allometric, and to examine the relationship between plant allometric growth and life history strategies in the spectrum. METHODS: Leaf and stem characters-including leaf and stem mass, total leaf area, individual leaf area, stem cross-sectional area, leaf number and stem length-at the twig level for 59 woody species were investigated along an altitudinal gradient on Changbaishan Mountain in the temperate zone of China. The environmental gradient ranges from temperate broad-leaved mixed forest at low altitude, to conifer forest at middle altitude, and to sub-alpine birch forest at high altitude. The scaling relationships between stem cross-sectional area and stem mass, stem mass and leaf mass, and leaf mass and leaf area at the twig level were simultaneously determined. KEY RESULTS: Twig cross-sectional area was found to have invariant allometric scaling relationships with the stem mass, leaf mass, total leaf area and individual leaf area, all with common slopes being significantly larger than 1, for three altitudinal-zoned vegetation types under investigation. However, leaf mass was found to be isometrically related to stem mass and leaf area along the environmental gradient. Based on the predictions of previous models, the exponent value of the relationship between twig cross sectional area and total leaf area can be inferred to be 1.5, which falls between the confidence intervals of the relationship at each altitude, and between the confidence intervals of the common slope value (1.17-1.56) of this study. This invariant scaling relationship is assumed to result from the fractural network and/or developmental constraints of plants. The allometric constants (y intercepts) of the relationships between the stem cross-sectional area and leaf area (both total leaf area and individual leaf area) were found to decrease significantly along the altitudinal gradient. This suggests that the species would support less leaf area at a given twig cross-sectional area with increasing environmental stress. CONCLUSIONS: This study demonstrated that plants respond to the environmental gradient by changing the y-intercepts of the relationship between leaf size-twig size, while keeping the exponent value of the allometric relationship as an invariant constant. The allometric growth in the twig size leaf size spectrum is related to many other components of plant life history strategy, including the well established life history trade-off between efficiency and safety in the hydraulic transport of water. PMID- 16254020 TI - A comparison between the record height-to-stem diameter allometries of Pachycaulis and Leptocaulis species. AB - BACKGROUND AND AIMS: The interspecific allometry of maximum plant height (Hmax) with respect to maximum basal stem diameter (Dmax) has been studied for leptocaulis dicot and conifer tree species. In contrast, virtually nothing is known about the interspecific allometry of pachycaulis species. Here, the interspecific allometries for palms, cacti and cycads are reported and compared with those of leptocaulis dicot and conifer tree species to determine whether pachycauly limits Hmax with respect to Dmax. METHODS: Data for each of a total of 1461 pachycaulis and leptocaulis species were gathered from the primary literature. The scaling exponent and the allometric constant of logHmax vs. logDmax reduced major axis regression curves (and their respective 95 % confidence intervals) were used to compare the four species groups. The stem slenderness ratio (Hmax/Dmax = Rmax) for each species was also computed to compare interspecific trends in trunk shape. KEY RESULTS AND CONCLUSIONS: Each of the four species groups is allometrically unique, i.e. no single 'canonical' maximum plant height to stem diameter allometry exists across all four species groups. Although pachycaulis does not intrinsically limit height, height is nevertheless limited by the size range of basal stem diameter occupied by each species group. Pachycaulis species achieve heights comparable to some leptocaulis species by virtue of very high slenderness ratios attended by an absence or paucity of stem branching. The diversity observed for pachycaulis stem allometries is likely the result of the independent evolutionary origins of this growth habit and the different anatomical strategies used to stiffen stems. PMID- 16254021 TI - Wound-induced oxidative responses in mountain birch leaves. AB - AIMS: The aim of the study was to examine oxidative responses in subarctic mountain birch, Betula pubescens subsp. czerepanovii, induced by herbivory and manual wounding. METHODS: Herbivory-induced changes in polyphenoloxidase, peroxidase and catalase activities in birch leaves were determined. A cytochemical dye, 3,3-diaminobenzidine, was used for the in situ and in vivo detection of H2O2 accumulation as a response to herbivory and wounding. To localize peroxidase activity in leaves, 10 mm H2O2 was applied to the dye reagent. KEY RESULTS: Feeding by autumnal moth, Epirrita autumnata, larvae caused an induction in polyphenoloxidase and peroxidase activities within 24 h, and a concomitant decrease in the activity of antioxidative catalases in wounded leaves. Wounding also induced H2O2 accumulation, which may have both direct and indirect defensive properties against herbivores. Wound sites and guard cells showed a high level of peroxidase activity, which may efficiently restrict invasion by micro-organisms. CONCLUSION: Birch oxidases together with their substrates may form an important front line in defence against herbivores and pathogens. PMID- 16254023 TI - Single agent carboplatin for CS IIA/B testicular seminoma. A phase II study of the German Testicular Cancer Study Group (GTCSG). AB - BACKGROUND: The aim was to investigate the use of single agent carboplatin in patients with seminoma stage IIA/B. PATIENTS AND METHODS: In a prospective phase II trial, single agent carboplatin at a dose of AUC 7 mg.min/ml every 4 weeks for three cycles in stage IIA (n=51) or four cycles in stage IIB (n=57) was given to 108 patients with previously untreated seminoma stage IIA/B. Patients with residual masses of >or=3 cm were scheduled to receive secondary surgery. RESULTS: A complete response (CR) was achieved by 88/108 (81%) patients, 17/108 (16%) achieved a partial response (PR), two of 108 (2%) showed no change, and one patient progressed. In all patients with PR the residual disease was or = 40 years. CONCLUSION: Goserelin given before and during chemotherapy may prevent premature menopause in the majority of patients. The different success rate by age, however, indicates the need of a prospective evidence of the efficacy of such a strategy. PMID- 16254022 TI - Epigenetics and its implications for plant biology. 1. The epigenetic network in plants. AB - BACKGROUND: Epigenetics has rapidly evolved in the past decade to form an exciting new branch of biology. In modern terms, 'epigenetics' studies molecular pathways regulating how the genes are packaged in the chromosome and expressed, with effects that are heritable between cell divisions and even across generations. CONTEXT: Epigenetic mechanisms often conflict with Mendelian models of genetics, and many components of the epigenetic systems in plants appeared anomalous. However, it is now clear that these systems govern how the entire genome operates and evolves. SCOPE: In the first part of a two-part review, how epigenetic systems in plants were elucidated is addressed. Also there is a discussion on how the different components of the epigenetic system--regulating DNA methylation, histones and their post-translational modification, and pathways recognizing aberrant transcripts--may work together. PMID- 16254025 TI - Fetal pituitary gonadotropin as an initial target of dioxin in its impairment of cholesterol transportation and steroidogenesis in rats. AB - Reproductive and developmental disorders are the most sensitive toxic effects caused by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). TCDD is thought to produce many, if not all, of these toxic effects by impairing steroidogenesis and/or steroid action during the prenatal or early postnatal stages. However, the mechanism of the antisex steroid effect of TCDD is not well understood. This study revealed that steroidogenic acute-regulatory protein (StAR), a key transporter of cholesterol for steroidogenesis, in the testes of fetal rats are down-regulated by maternal exposure to TCDD. It was also shown that many mRNAs of steroidogenetic enzymes, including cytochromes P450 11A1, 17, and 11B1 and 3beta hydroxysteroid dehydrogenase, are reduced in fetuses of TCDD-treated dams in a testis-specific manner. The same was also observed for the expression of estrogen alpha receptors and androgen receptors. Whereas StAR expression was not affected by TCDD in cultured fetal testis, the fetal serum content of LH, a pituitary regulator of StAR, was significantly reduced by TCDD. In agreement with this, pituitary expression of LHbeta subunit mRNA in fetuses was reduced by maternal exposure to TCDD, whereas the alpha-subunit remained unchanged. The reduction in LHbeta is suggested to occur by a mechanism different from the reduction in the GnRH level. Direct supply of exogenous gonadotropin to TCDD-exposed fetuses completely abolished the reduction of StAR expression. Taken together, these results demonstrate that TCDD impairs steroidogenesis in the fetus by targeting pituitary gonadotropins. PMID- 16254026 TI - Both early and delayed treatment with melanocortin 4 receptor-stimulating melanocortins produces neuroprotection in cerebral ischemia. AB - Ischemic stroke is one of the main causes of death and disability. We investigated whether melanocortin peptides, which have protective effects in severe hypoxic conditions, also produce neuroprotection in a gerbil model of ischemic stroke. A 10-min period of global cerebral ischemia, induced by occluding both common carotid arteries, caused impairment in spatial learning and memory that was associated with activation of inflammatory and apoptotic pathways, including severe DNA damage and delayed neuronal death, in the hippocampus. Treatment with nanomolar doses of the melanocortin analog [Nle4, D Phe7] alpha-MSH [which activates the melanocortin receptor subtypes (MC) mainly expressed in central nervous system, namely MC3 and MC4] modulated the inflammatory and apoptotic cascades and reduced hippocampus injuries even when delayed up to 9 h after ischemia, with consequent dose-dependent improvement in subsequent functional recovery. The selective MC3 receptor agonist gamma2-MSH had no protective effects. Pharmacological blockade of MC4 receptors prevented the neuroprotective effects of [Nle4, D-Phe7] alpha-MSH and worsened some ischemia outcomes. Together, our findings suggest that MC4 receptor-stimulating melanocortins might provide potential to develop a class of drugs with a broad treatment window for a novel approach to neuroprotection in ischemic stroke. PMID- 16254027 TI - Estrogen regulates transcription of the ovine oxytocin receptor gene through GC rich SP1 promoter elements. AB - Establishment of pregnancy in ruminants results from paracrine signaling by interferon tau (IFNT) from the conceptus to uterine endometrial luminal epithelia (LE) that prevents release of luteolytic prostaglandin F(2alpha) pulses. In cyclic and pregnant ewes, progesterone down-regulates progesterone receptor (PGR) gene expression in LE. In cyclic ewes, loss of PGR allows for increases in estrogen receptor alpha (ESR1) and then oxytocin receptor (OXTR) gene expression followed by oxytocin-induced prostaglandin F(2alpha) pulses. In pregnant ewes, IFNT inhibits transcription of the ESR1 gene, which presumably inhibits OXTR gene transcription. Alternatively, IFNT may directly inhibit OXTR gene transcription. The 5' promoter/enhancer region of the ovine OXTR gene was cloned and found to contain predicted binding sites for activator protein 1, SP1, and PGR, but not for ESR1. Deletion analysis showed that the basal promoter activity was dependent on the region from -144 to -4 bp that contained only SP1 sites. IFNT did not affect activity of the OXTR promoter. In cells transfected with ESR1, E2, and ICI 182,780 increased promoter activity due to GC-rich SP1 binding sites at positions -104 and -64. Mutation analyses showed that the proximal SP1 sites mediated ESR1 action as well as basal activity of the promoter. In response to progesterone, progesterone receptor B also increased OXTR promoter activity. SP1 protein was constitutively expressed and abundant in the LE of the ovine uterus. These results support the hypothesis that the antiluteolytic effects of IFNT are mediated by direct inhibition or silencing of ESR1 gene transcription, thereby precluding ESR1/SP1 from stimulating OXTR gene transcription. PMID- 16254028 TI - Endogenous relaxin regulates collagen deposition in an animal model of allergic airway disease. AB - We examined the relationship among relaxin (a peptide hormone that stimulates collagen degradation), airway fibrosis, other changes of airway remodeling, and airway hyperresponsiveness (AHR) in an animal model of allergic airway disease. Eight- to 10-wk-old relaxin gene-knockout (RLX(-/-)) and wild-type (RLX(+/+)) mice were sensitized with ovalbumin (OVA) or saline ip at d 0 and 14 and challenged three times per week for 6 wk with nebulized 2.5% OVA or saline. Saline-treated control RLX(+/+) and RLX(-/-) mice had equivalent collagen expression and baseline airway responses. OVA-treated RLX(-/-) mice developed airway inflammation equivalent to that in OVA-treated RLX(+/+) mice. However, OVA treated RLX(-/-) mice had markedly increased lung collagen deposition as compared with OVA-treated RLX(+/+) and saline-treated mice (all P < 0.05). Collagen was predominantly deposited in the subepithelial basement membrane region and submucosal regions in both OVA-treated RLX(+/+) and RLX(-/-) mice. The increased collagen measured in OVA-treated RLX(-/-) mice was associated with reduced matrix metalloproteinase (MMP)-9 (P < 0.02) expression and failure to up-regulate matrix metalloproteinase-2 expression, compared with levels in OVA-treated RLX(+/+) mice. Goblet cell numbers were equivalent in OVA-treated RLX(-/-) and RLX(+/+) mice and increased, compared with saline-treated animals. Both OVA-treated RLX(+/+) and RLX(-/-) mice developed similar degrees of AHR after OVA treatment. These findings demonstrate a critical role for relaxin in the inhibition of lung collagen deposition during an allergic inflammatory response. Increased deposition of collagen per se did not influence airway epithelial structure or AHR. PMID- 16254030 TI - Ectopic expression of the gastric inhibitory polypeptide receptor gene is a sufficient genetic event to induce benign adrenocortical tumor in a xenotransplantation model. AB - Aberrant expression of ectopic G protein-coupled receptors (GPCRs) in adrenal cortex tissue has been observed in several cases of ACTH-independent macronodular adrenal hyperplasias and adenomas associated with Cushing's syndrome. Although there is clear clinical evidence for the implication of these ectopic receptors in abnormal regulation of cortisol production, whether this aberrant GPCR expression is the cause or the consequence of the development of an adrenal hyperplasia is still an open question. To answer it, we genetically engineered primary bovine adrenocortical cells to have them express the gastric inhibitory polypeptide receptor. After transplantation of these modified cells under the renal capsule of adrenalectomized immunodeficient mice, tissues formed had their functional and histological characteristics analyzed. We observed the formation of an enlarged and hyperproliferative adenomatous adrenocortical tissue that secreted cortisol in a gastric inhibitory polypeptide-dependent manner and induced a mild Cushing's syndrome with hyperglycemia. Moreover, we show that tumor development was ACTH independent. Thus, a single genetic event, inappropriate expression of a nonmutated GPCR gene, is sufficient to initiate the complete phenotypic alterations that ultimately lead to the formation of a benign adrenocortical tumor. PMID- 16254031 TI - Nongenomic action of progesterone inhibits oxytocin-induced phosphoinositide hydrolysis and prostaglandin F2alpha secretion in the ovine endometrium. AB - Experiments were conducted to characterize the nongenomic effects of progesterone (P4) on binding of oxytocin (OT) to its receptor and signal transduction in the ovine endometrium. The dose-response relationship of P4 to OT binding was examined. Membranes from endometrial tissue of ovariectomized hormone-treated ewes were preincubated in the presence of P4 for 1 h followed by OT receptor analysis. P4 interfered with the binding of OT in a dose-dependent manner. Endometrium was then recovered from cyclic ewes and divided into explants. Treatment consisted of two dosages of P4 and two dosages of OT. Explants were analyzed for total inositol monophosphate, bisphosphate (IP(2)), and trisphosphate (IP(3)) content. Preincubation with P4 for 10 min significantly interfered with OT stimulation of IP(2) and IP(3) synthesis. Oxytocin increased monophosphate production, but there was no detectable effect of P4. In the next experiment, endometrial explants were cultured in the absence or the presence of arachidonic acid. Explants were then exposed for 1 h to medium containing vehicle or P4. After incubation, explants were challenged with OT and the media were collected and analyzed for 13,14 dihydro-15-keto prostaglandin F(2alpha) by RIA. Treatment of explants with AA increased PGF(2alpha) content compared with that of controls. Brief exposure to P4 significantly decreased OT-induced PGF(2alpha) secretion from explants previously exposed to medium or AA. Collectively, these data are interpreted to indicate that the observed reduction in OT-induced IP(2) and IP(3) production and OT-induced PGF(2alpha) secretion was due to P4 inhibition of OT binding to its receptor. PMID- 16254029 TI - Tumor necrosis factor-alpha activates the human prolactin gene promoter via nuclear factor-kappaB signaling. AB - Pituitary function has been shown to be regulated by an increasing number of intrapituitary factors, including cytokines. Here we show that the important cytokine TNF-alpha activates prolactin gene transcription in pituitary GH3 cells stably expressing luciferase under control of 5 kb of the human prolactin promoter. Similar regulation of the endogenous rat prolactin gene by TNF-alpha in GH3 cells was confirmed using real-time PCR. Luminescence microscopy revealed heterogeneous dynamic response patterns of promoter activity in individual cells. In GH3 cells treated with TNF-alpha, Western blot analysis showed rapid inhibitory protein kappaB (IkappaBalpha) degradation and phosphorylation of p65. Confocal microscopy of cells expressing fluorescence-labeled p65 and IkappaBalpha fusion proteins showed transient cytoplasmic-nuclear translocation and subsequent oscillations in p65 localization and confirmed IkappaBalpha degradation. This was associated with increased nuclear factor kappaB (NF-kappaB)-mediated transcription from an NF-kappaB-responsive luciferase reporter construct. Disruption of NF-kappaB signaling by expression of dominant-negative variants of IkappaB kinases or truncated IkappaBalpha abolished TNF-alpha activation of the prolactin promoter, suggesting that this effect was mediated by NF-kappaB. TNF alpha signaling was found to interact with other endocrine signals to regulate prolactin gene expression and is likely to be a major paracrine modulator of lactotroph function. PMID- 16254032 TI - Differential endocrine responses to rosiglitazone therapy in new mouse models of type 2 diabetes. AB - Polygenic mouse models for obesity-induced type 2 diabetes (T2D) more accurately reflect the most common manifestations of the human disease. Two inbred mouse strains (NON/Lt and NZO/HlLt) separately contributed T2D susceptibility- conferring quantitative trait loci to F1 males. Although chronic administration of rosiglitazone (Rosi) in diet (50 mg/kg) effectively suppressed F1 diabetes, hepatosteatosis was an undesired side effect. Three recombinant congenic strains (designated RCS1, -2, and -10) developed on the NON/Lt background carry variable numbers of these quantitative trait loci that elicit differential weight gain and male glucose intolerance syndromes of variable severity. We previously showed that RCS1 and -2 mice responded to chronic Rosi therapy without severe steatosis, whereas RCS10 males were moderately sensitive. In contrast, another recombinant congenic strain, RCS8, responded to Rosi therapy with the extreme hepatosteatosis observed in the F1. Longitudinal changes in multiple plasma analytes, including insulin, the adipokines leptin, resistin, and adiponectin, and plasminogen activator inhibitor-1 (PAI-1) allowed profiling of the differential Rosi responses in steatosis-exacerbated F1 and RCS8 males vs. the resistant RCS1 and RCS2 or moderately sensitive RCS10. Of these biomarkers, PAI-1 most effectively predicted adverse drug responses. Unexpectedly, mean resistin concentrations were higher in Rosi-treated RCS8 and RCS10. In summary, longitudinal profiling of multiple plasma analytes identified PAI-1 as a useful biomarker to monitor for differential pharmacogenetic responses to Rosi in these new mouse models of T2D. PMID- 16254033 TI - Bilirubin can induce tolerance to islet allografts. AB - Induction of heme oxygenase-1 (HO-1) expression in recipients of allogeneic islets can lead to long-term survival (>100 d) of those islets. We tested whether administration of bilirubin would substitute for the beneficial effects of HO-1 expression in islet transplantation. Administering bilirubin to the recipient (B6AF1) or incubating islets in a bilirubin-containing solution ex vivo led to long-term survival of allogeneic islets in a significant percentage of cases. In addition, administering bilirubin to only the donor frequently led to long-term survival of DBA/2 islets in B6AF1 recipients and significantly prolonged graft survival of BALB/c islets in C57BL/6 recipients. Donor treatment with bilirubin up-regulated mRNA expression of protective genes such as HO-1 and bcl-2 and suppressed proinflammatory and proapoptotic genes including monocyte chemoattractant protein-1 and caspase-3 and -8 in the islet grafts before transplantation. Furthermore, treatment of only the donor suppressed the expression of proinflammatory cytokines including TNF-alpha, inducible nitric oxide synthase, monocyte chemoattractant protein-1, and other proapoptotic and proinflammatory genes normally seen in the islets after transplantation. Donor treatment also reduced the number of macrophages that infiltrated the islet grafts in the recipients. Preincubation of betaTC3 cells with bilirubin also protected the cells from lipid peroxidation. Our data suggests that the potent antioxidant and antiinflammatory actions of bilirubin may contribute to islet survival. PMID- 16254034 TI - Inhibition of 11beta-hydroxysteroid dehydrogenase eliminates impaired glucocorticoid suppression and induces apoptosis in corticotroph tumor cells. AB - Cushing's disease is characterized by persistent ACTH secretion under hypercortisolemia. In an attempt to clarify the molecular mechanism, we examined the effect of 11beta-hydroxysteroid dehydrogenase (HSD) inhibition on glucocorticoid suppression of ACTH release using murine corticotroph tumor cells. We found that 11beta-HSD2, as well as -HSD1, was expressed in the cells and that its inhibition by carbenoxolone significantly improved the negative feedback effect of glucocorticoid. Carbenoxolone also enhanced apoptosis induced by cortisol. These effects are most likely attributable to inhibition of 11beta-HSD2 because only cortisol, a substrate of 11beta-HSD2, was present in these experimental conditions. We conclude that ectopic expression of 11beta-HSD2 is, at least in part, responsible for the impaired glucocorticoid suppression in corticotroph adenoma. Inhibition of 11beta-HSD2 may be applicable to the medical therapy for Cushing's disease. PMID- 16254035 TI - The progesterone receptor in human term amniochorion and placenta is isoform C. AB - The mechanism that initiates human parturition has been proposed to be functional progesterone withdrawal whereby the 116-kDa B isoform of the progesterone receptor (PR-B) switches in favor of the 94-kDa A isoform (PR-A) in reproductive tissues. Recently other PR isoforms, PR-S, PR-C, and PR-M generated from the same gene have been identified and partially characterized. Using immunohistochemical, Western blotting, and RT-PCR techniques, evidence is provided that the major PR isoform present in human term fetal membranes (amnion and chorion) and syncytiotrophoblast of the placenta is neither of the classical nuclear PR-B or PR-A isoforms but is the N terminally truncated 60-kDa PR-C isoform. Evidence is also provided that the PR-C isoform resides in the cytoplasm of the expressing cell types. Data are also presented to show that PR-B, PR-A, and PR-S isoforms are essentially absent from the amnion and chorion, whereas PR isoforms A, B, C, and S are all present in the decidua, with PR-A being the major isoform. The syncytiotrophoblast of the placenta contains the cytoplasmic PR-C isoform but not PR-A, PR-B, or PR-S. The major PR isoform in the amnion, chorion, and placenta is PR-C, suggesting that the cytoplasmic PR-C isoform has a specific role in extraembryonic tissues and may be involved in the regulation of human parturition. PMID- 16254036 TI - BRAF mediates RET/PTC-induced mitogen-activated protein kinase activation in thyroid cells: functional support for requirement of the RET/PTC-RAS-BRAF pathway in papillary thyroid carcinogenesis. AB - In human papillary thyroid cancers (PTCs), mutations of RET/PTC, NTRK, RAS, or BRAF are found in about two thirds of cases with practically no overlap, providing genetic evidence that constitutive signaling along RET-RAS-BRAF-MAPK is key to their development. The requirement for BRAF in RET/PTC-mediated MAPK activation and gene expression has not been tested functionally. There are three RAF isoforms: ARAF, BRAF, and CRAF. Compared with the others, ARAF is a much weaker stimulator of MAPK. To determine the key RAF isoform mediating RET/PTC induced ERK phosphorylation, we stably transfected doxycycline-inducible RET/PTC3 expressing thyroid PCCL3 cells with small interfering RNA vectors to induce selective knockdown of BRAF or CRAF. Conditional RET/PTC3 expression induced comparable ERK phosphorylation in CRAF knockdown and control cells but negligible ERK phosphorylation in BRAF knockdown cells. Selective knockdown of BRAF prevented RET/PTC-dependent down-regulation of the sodium iodide symporter, a gene that confers key biological effects of RET/PTC in PTCs. Moreover, microarray analysis revealed numerous RET/PTC-regulated genes showing requirement of BRAF for appropriate expression. These data indicate that BRAF is required for RET/PTC induced MAPK activation in thyroid cells and support the notion that BRAF inactivation may be an attractive target for PTCs. PMID- 16254037 TI - Reduction in voltage-gated K+ currents in primary cultured rat pancreatic beta cells by linoleic acids. AB - Free fatty acids (FFAs), in addition to glucose, have been shown to stimulate insulin release through the G protein-coupled receptor (GPCR)40 receptor in pancreatic beta-cells. Intracellular free calcium concentration ([Ca(2+)](i)) in beta-cells is elevated by FFAs, although the mechanism underlying the [Ca(2+)](i) increase is still unknown. In this study, we investigated the action of linoleic acid on voltage-gated K(+) currents. Nystatin-perforated recordings were performed on identified rat beta-cells. In the presence of nifedipine, tetrodotoxin, and tolbutamide, voltage-gated K(+) currents were observed. The transient current represents less than 5%, whereas the delayed rectifier current comprises more than 95%, of the total K(+) currents. A long-chain unsaturated FFA, linoleic acid (10 microm), reversibly decreased the amplitude of K(+) currents (to less than 10%). This reduction was abolished by the cAMP/protein kinase A system inhibitors H89 (1 microm) and Rp-cAMP (10 microm) but was not affected by protein kinase C inhibitor. In addition, forskolin and 8'-bromo-cAMP induced a similar reduction in the K(+) current as that evoked by linoleic acid. Insulin secretion and cAMP accumulation in beta-cells were also increased by linoleic acid. Methyl linoleate, which has a similar structure to linoleic acid but no binding affinity to GPR40, did not change K(+) currents. Treatment of cultured cells with GPR40-specific small interfering RNA significantly reduced the decrease in K(+) current induced by linoleic acid, whereas the cAMP-induced reduction of K(+) current was not affected. We conclude that linoleic acid reduces the voltage-gated K(+) current in rat beta-cells through GPR40 and the cAMP-protein kinase A system, leading to an increase in [Ca(2+)](i) and insulin secretion. PMID- 16254038 TI - Expression of tumor suppressor and tumor-related proteins in differentiated carcinoma, undifferentiated carcinoma with tubular component and pure undifferentiated carcinoma of the stomach. AB - BACKGROUND: The recent development of tissue microarray (TMA) technology allows high-throughput protein expression profiling of cancer tissues by immunohistochemistry. We attempted to clarify the derivation of undifferentiated type gastric carcinoma with tubular component by using TMA. METHODS: We constructed a TMA system composed of six paraffin blocks in which 274 samples of formalin-fixed gastric carcinoma tissue from 274 patients were embedded. Using this system, we performed immunohistochemical stains for five tumor suppressor and tumor-related proteins, i.e. p53, p16, hMLH1, c-erbB-2 and carcinoembryonic antigen (CEA). The 274 gastric carcinomas were histopathologically divided into the following three groups according to the degree of differentiation: differentiated-type (D-type), undifferentiated-type with tubular component (UT type) and pure undifferentiated-type (UP-type). Immunohistochemical results were then compared with histological types. RESULTS: The percentages of abnormal expression of each protein in D-type, UT-type and UP-type carcinomas were as follows: 27% (38/143), 17% (17/98) and 15% (5/33) for p53; 27% (39/143), 19% (19/98) and 18% (6/33) for p16; 38% (54/143), 44% (43/98) and 24% (8/33) for hMLH1; 15% (22/143), 5% (5/98) and 0% (0/33) for c-erbB-2; and 22% (31/143), 35% (34/98) and 70% (23/33) for CEA. UP-type carcinomas exhibited the lowest frequencies of abnormal expression for p53, p16, hMLH1 and c-erbB-2, but the highest frequencies for CEA. UT-type carcinomas generally showed intermediate frequencies between those of D-type and UP-type carcinomas. Differences between D type and UP-type for c-erbB-2 (P < 0.05) and CEA (P < 0.001) were significant, as were differences between D-type and UT-type for c-erbB-2 (P < 0.05) and CEA (P < 0.05), and differences between UT-type and UP-type for hMLH1 (P < 0.05) and CEA (P < 0.001). CONCLUSIONS: These findings reveal that gastric carcinomas have distinct expression profiles for tumor suppressor and tumor-related proteins depending on histological types, and support the hypothesis that UT-type carcinomas are derived not only from D-type but also from UP-type carcinomas. We also found significant differences between abnormal protein expression and other clinicopathological parameters such as gender, age and status of tumor and nodes. PMID- 16254039 TI - Synchronous adenocarcinoma and mucosa-associated lymphoid tissue (MALT) lymphoma in a single stomach. AB - BACKGROUND: Synchronous gastric adenocarcinoma and primary gastric mucosa associated lymphoid tissue (MALT) lymphoma are rare, and the characteristics of these tumors are still unclear. METHODS: Histology of endoscopic and surgical specimens of 6012 gastric adenocarcinoma and 25 primary gastric MALT lymphoma cases were reviewed. RESULTS: Five cases of synchronous tumors were found among 25 primary gastric MALT lymphoma patients (20.0%) and among 6012 gastric adenocarcinoma patients (8.3%). Helicobacter pylori was detected only in two cases (40.0%) of these five synchronous cases. In only one case, two types of malignant cells were mingled each other as a colliding tumor. In other four cases, two pathologies coexisted separately. These two types of malignant tumors were not distinguished by endoscopic or gross findings. CONCLUSIONS: Although gastric adenocarcinoma coexists with MALT lymphoma at a relatively high frequency, synchronous carcinoma can be misinterpreted as a multifocal separated lesion of MALT lymphoma owing to their macroscopic resemblance. Therefore, gastric MALT lymphoma patients should be carefully examined by endoscopy, and any suspicious area must be biopsied considering the possibility of coexisting adenocarcinoma. In addition, etiologies other than H. pylori should be considered in these synchronous tumors. PMID- 16254040 TI - Ovarian cyst formation in patients using tamoxifen for breast cancer. AB - OBJECTIVE: The purpose of this study was to evaluate patient-related parameters that determine ovarian cyst formation in women using tamoxifen for breast cancer. METHODS: A retrospective review of tamoxifen-treated women with breast cancer who were followed up in the outpatient clinic at Ankara Oncology Hospital between January 2002 and December 2004 was performed. Tamoxifen doses and duration, post treatment menstrual function, adjuvant therapy, ultrasonographic and hormonal [follicle-stimulating hormone and serum estradiol (E(2))] data, details of gynecologic surgical procedure and histopathology were recorded. RESULTS: Twenty nine of 150 tamoxifen-treated patients (19.3%) had ovarian cysts. Cysts were detected in 28 of 57 pre-menopausal women (49.1%) and 1 of 93 post-menopausal women (1.1%). Patients with ovarian cysts had higher serum E(2) levels compared with patients without cysts (24 versus 345 pg/ml; P < 0.001). Patients with ovarian cysts had <1 year amenorrhoea duration (P < 0.001) compared with the patients without cysts. Adjuvant standard chemotherapy did not have relationship between the development of ovarian cysts. Multivariant analysis showed that cyst development is related to high E(2) levels (P < 0.05). CONCLUSIONS: Patients still having a menstrual cycle during tamoxifen had high risk (58.33%) of developing ovarian cysts. We have described an association between pre-menopausal patients using tamoxifen with high E(2) level and ovarian cyst enlargement. PMID- 16254041 TI - Comparison of laryngeal cancer mortality in five countries: France, Italy, Japan, UK and USA from the WHO mortality database (1960-2000). PMID- 16254042 TI - Nitric oxide down-regulates the expression of the catalytic NADPH oxidase subunit Nox1 in rat renal mesangial cells. AB - Glomerular mesangial cells can produce high amounts of nitric oxide (NO) and reactive oxygen species (ROS). Here we analyzed the impact of NO on the ROS generating system, particularly on the NADPH oxidase Nox1. Nox1 mRNA and protein levels were markedly decreased by treatment of mesangial cells with the NO releasing compound DETA-NO in a concentration- and time-dependent fashion. By altering the cGMP signaling system with different inhibitors or activators, we revealed that the effect of NO on Nox1 expression is at least in part mediated by cGMP. Analysis of a reporter construct comprising the 2547 bp of the nox1 promoter region revealed that a stimulatory effect of IL-1beta on nox1 transcription is counteracted by an inhibitory effect of IL-1beta-evoked endogenous NO formation. Moreover, pretreatment of mesangial cells with DETA-NO attenuated platelet-derived growth factor (PDGF)-BB or serum stimulated production of superoxide as assessed by real-time EPR spectroscopy and dichlorofluorescein formation. Transfection of mesangial cells with siRNAs directed against Nox1 and Nox4 revealed that inhibition of Nox1, but not Nox4 expression, is responsible for the reduced ROS formation by NO. Obviously, there exists a fine-tuned crosstalk between NO and ROS generating systems in the course of inflammatory diseases. PMID- 16254043 TI - Impaired fatty acid utilization in thioredoxin binding protein-2 (TBP-2) deficient mice: a unique animal model of Reye syndrome. AB - Thioredoxin binding protein-2 (TBP-2) is a negative regulator of thioredoxin and has multiple regulatory functions in cellular redox, growth, differentiation, apoptosis, and aging. To investigate the function of TBP-2 in vivo, we generated mice with targeted inactivation of TBP-2 (TBP-2-/- mice). Here, we show that TBP 2 expression is markedly up-regulated during fasting in wild-type mice, while TBP 2-/- mice were predisposed to death with bleeding tendency, as well as hepatic and renal dysfunction as a result of 48 h of fasting. The fasting-induced death was rescued by supplementation of glucose but not by that of oleic acid, suggesting that inability of fatty acid utilization plays an important role in the anomaly of TBP-2-/- mice. In these mice, plasma free fatty acids levels are higher, whereas glucose levels are lower than those of wild-type mice. Compared with wild-type mice, TBP-2-/- mice showed increased levels of plasma ketone bodies, pyruvate and lactate, indicating that Krebs cycle-mediated fatty acid utilization is impaired. Because the fatal impairment of fatty acid utilization is a characteristically metabolic feature of Reye (-like) syndrome, TBP-2-/- mouse may represent a novel model for investigating the pathophysiology of these disorders. PMID- 16254044 TI - Regulation of store-operated calcium entries and mitochondrial uptake by minidystrophin expression in cultured myotubes. AB - Defective expression of dystrophin in muscle cells is the primary feature of Duchenne muscular dystrophy (DMD), which is accompanied by fiber necrosis and intracellular calcium mishandling. These features led to the hypothesis that dystrophin could control calcium movements. Calcium mishandling in human DMD myotubes is dependent on contraction and/or calcium release activity, suggesting the involvement of channels being activated during these processes. Forced expression of minidystrophin at the plasma membrane of dystrophin-deficient Sol8 myotubes reactivates appropriate sarcolemmal expression of dystrophin-associated proteins and results in normal calcium homeostasis. In active dystrophic myotubes, store-operated calcium channels could be responsible for a sustained calcium influx in muscle cells. We show here that depletion of calcium stores (sarcoplasmic reticulum) by repetitive activation of calcium release and blockade of SERCA leads to a calcium influx. In myotubes expressing recombinant minidystrophin, these store-dependent influxes were reduced to a level similar to that observed in myotubes expressing native dystrophin. High store-dependent calcium influxes in dystrophin-deficient myotubes were associated with sustained cytosolic calcium transients and high intramitochondrial entries, while lower store-dependent calcium influx in myotubes expressing minidystrophin resulted in shorter calcium transients and reduced calcium uptake into mitochondria. We propose that minidystrophin negatively regulates sarcolemmal store-dependent calcium channels, which reduces store-dependent calcium influx, as well as its mitochondrial uptake. Forced expression of minidystrophin in dystrophic cells might restore the regulation of sarcolemmal store-dependent channels, which could protect against calcium mishandling. PMID- 16254045 TI - A cell-based high-throughput assay for screening inhibitors of human papillomavirus-16 long control region activity. AB - Cervical carcinomas express human papillomavirus (HPV) E6 and E7 oncoproteins, which are required to maintain the proliferative state of cancer cells. Repression of E6 and E7 expression results in acquisition of senescent phenotype and in the rescue of functional p53 and p105(Rb) pathways; therefore, therapies directed against either viral protein may be beneficial. However, the systems to study HPV in vitro are technically difficult and not convenient for screening of antiviral compounds. This has hampered the discovery of drugs against HPV. Here we describe the generation and use of a high-throughput screening system based on keratinocytes stably transfected with a reporter construct containing the regulatory sequence of E6 and E7 gene transcription (LCR) that allows easy detection of inhibitors of E6 and E7 expression in libraries of synthetic or biological compounds. The assay was used to screen a wide panel of cytokines: among them, IL-4, IL-13, TGF-beta1, TGF-beta2, TGF-beta3, TNF-alpha, IFN-alpha, and IFN-beta were found to induce a strong inhibition of the LCR activity. Our assay provides a validated tool in the search for drugs against HPV-associated cervical carcinomas and allowed the first systematic analysis of the effect of cytokines on the HPV-16 LCR transcriptional activity. PMID- 16254046 TI - Involvement of cyclooxygenase-2 in gastric mucosal hypertrophy in gastrin transgenic mice. AB - Gastrin promotes gastric mucosal growth, and hypergastrinemia induces gastric mucosal hypertrophy. Recently, it has been reported that gastrin induces cyclooxygenase-2 (COX-2) in human gastric and colorectal cancer cell lines. However, whether COX-2 is involved in gastrin-induced gastric mucosal growth in vivo is unknown. We investigated the role of COX-2 in gastrin-induced gastric mucosal hypertrophy using gastrin transgenic mice. Hypergastrinemic mice [mice with mutated gastrin under the control of the beta-actin promoter (ACT-GAS mice)] received the COX-2 inhibitor celecoxib (0, 200, or 500 mg/kg of diet) from 5 wk of age and were killed at 16 or 24 wk. Some ACT-GAS mice received celecoxib from 16 wk and were killed at 24 wk. Eighty-week-old ACT-GAS mice without celecoxib treatment were also examined. The thickness of the gastric mucosa, cell populations, COX-2 expression, and PGE(2) levels were evaluated. All ACT-GAS mice showed gastric mucosal hypertrophy, and four of six 80-wk-old ACT-GAS mice developed gastric cancer. COX-2 was expressed in interstitial cells of the hypertrophic gastric mucosa and gastric cancers. Moreover, PGE(2) levels in the gastric mucosa of ACT-GAS mice were significantly higher than those of normal mice. With treatment with celecoxib, PGE(2) levels, the gastric mucosal thickness, and the number of total gastric cells per gastric gland of ACT-GAS mice were significantly decreased. The decrease in gastric mucosal thickness was caused by a reduction of foveolar hyperplasia. The thickness of glandules and the number of Ki67-positive cells were not significantly changed. In conclusion, COX 2 contributes to gastrin-induced mucosal hypertrophy of the stomach. PMID- 16254047 TI - Fatty acid transport and metabolism in HepG2 cells. AB - The mechanism(s) of fatty acid uptake by liver cells is not fully understood. We applied new approaches to address long-standing controversies of fatty acid uptake and to distinguish diffusion and protein-based mechanisms. Using HepG2 cells containing an entrapped pH-sensing fluorescence dye, we showed that the addition of oleate (unbound or bound to cyclodextrin) to the external buffer caused a rapid (seconds) and dose-dependent decrease in intracellular pH (pH(in)), indicating diffusion of fatty acids across the plasma membrane. pH(in) returned to its initial value with a time course (in min) that paralleled the metabolism of radiolabeled oleate. Preincubation of cells with the inhibitors phloretin or triacsin C had no effect on the rapid pH(in) drop after the addition of oleate but greatly suppressed pH(in) recovery. Using radiolabeled oleate, we showed that its esterification was almost completely inhibited by phloretin or triacsin C, supporting the correlation between pH(in) recovery and metabolism. We then used a dual-fluorescence assay to study the interaction between HepG2 cells and cis-parinaric acid (PA), a naturally fluorescent but slowly metabolized fatty acid. The fluorescence of PA increased rapidly upon its addition to cells, indicating rapid binding to the plasma membrane; pH(in) decreased rapidly and simultaneously but did not recover within 5 min. Phloretin had no effect on the PA-mediated pH(in) drop or its slow recovery but decreased the absolute fluorescence of membrane-bound PA. Our results show that natural fatty acids rapidly bind to, and diffuse through, the plasma membrane without hindrance by metabolic inhibitors or by an inhibitor of putative membrane-bound fatty acid transporters. PMID- 16254048 TI - Electroacupuncture at ST-36 accelerates colonic motility and transit in freely moving conscious rats. AB - Acupuncture is useful for functional bowel diseases, such as constipation and diarrhea. However, the mechanisms of beneficial effects of acupuncture on colonic function have scarcely ever been investigated. We tested the hypothesis that electroacupuncture (EA) at ST-36 stimulates colonic motility and transit via a parasympathetic pathway in conscious rats. Hook-shaped needles were inserted at bilateral ST-36 (lower limb) or BL-21 (back) and electrically stimulated at 10 Hz for 20 min. We also studied c-Fos expression in response to EA at ST-36 in Barrington's nucleus of the pons. EA at ST-36, but not BL-21, significantly increased the amplitude of motility at the distal colon. The calculated motility index of the distal colon increased to 132 +/- 9.9% of basal levels (n = 14, P < 0.05). In contrast, EA at ST-36 had no stimulatory effects in the proximal colon. EA at ST-36 significantly accelerated colonic transit [geometric center (GC) = 6.76 +/- 0.42, n = 9, P < 0.001] compared with EA at BL-21 (GC = 5.23 +/- 0.39, n = 7). The stimulatory effect of EA at ST-36 on colonic motility and transit was abolished by pretreatment with atropine. EA-induced acceleration of colonic transit was also abolished by extrinsic nerve denervation of the distal colon (GC = 4.69 +/- 0.33, n = 6). The number of c-Fos-immunopositive cells at Barrington's nucleus significantly increased in response to EA at ST-36 to 8.1 +/- 1.1 cells/section compared with that of controls (2.4 +/- 0.5 cells/section, n = 3, P < 0.01). It is concluded that EA at ST-36 stimulates distal colonic motility and accelerates colonic transit via a sacral parasympathetic efferent pathway (pelvic nerve). Barrington's nucleus plays an important role in mediating EA-induced distal colonic motility in conscious rats. PMID- 16254049 TI - Arp2/3 complex-deficient mouse fibroblasts are viable and have normal leading edge actin structure and function. AB - RNA interference silencing of up to 90% of Arp3 protein expression, a major subunit of the Arp2/3 complex, proportionately decreases the intracellular motility of Listeria monocytogenes and actin nucleation activity ascribable to the Arp2/3 complex in mouse embryonic fibroblasts. However, the Arp2/3-deficient cells exhibit unimpaired lamellipodial actin network structure, translational locomotion, spreading, actin assembly, and ruffling responses. In addition, Arp3 silenced cells expressing neural Wiskott-Aldrich syndrome protein-derived peptides that inhibit Arp2/3 complex function in wild-type cells retained normal PDGF-induced ruffling. The Arp2/3 complex can be dispensable for leading-edge actin remodeling. PMID- 16254050 TI - The eukaryotic initiation factor (eIF) 5 HEAT domain mediates multifactor assembly and scanning with distinct interfaces to eIF1, eIF2, eIF3, and eIF4G. AB - Eukaryotic translation initiation factor (eIF) 5 is crucial for the assembly of the eukaryotic preinitiation complex. This activity is mediated by the ability of its C-terminal HEAT domain to interact with eIF1, eIF2, and eIF3 in the multifactor complex and with eIF4G in the 48S complex. However, the binding sites for these factors on eIF5-C-terminal domain (CTD) have not been known. Here we present a homology model for eIF5-CTD based on the HEAT domain of eIF2Bepsilon. We show that the binding site for eIF2beta is located in a surface area containing aromatic and acidic residues (aromatic/acidic boxes), that the binding sites for eIF1 and eIF3c are located in a conserved surface region of basic residues, and that eIF4G binds eIF5-CTD at an interface overlapping with the acidic area. Mutations in these distinct eIF5 surface areas impair GCN4 translational control by disrupting preinitiation complex interactions. These results indicate that the eIF5 HEAT domain is a critical nucleation core for preinitiation complex assembly and function. PMID- 16254051 TI - Receptor-mediated glutamate release from volume sensitive channels in astrocytes. AB - Several lines of work have shown that astrocytes release glutamate in response to receptor activation, which results in a modulation of local synaptic activity. Astrocytic glutamate release is Ca(2+)-dependent and occurs in conjunction with exocytosis of glutamate containing vesicles. However, astrocytes contain a millimolar concentration of cytosolic glutamate and express channels permeable to small anions, such as glutamate. Here, we tested the idea that astrocytes respond to receptor stimulation by dynamic changes in cell volume, resulting in volume sensitive channel activation, and efflux of cytosolic glutamate. Confocal imaging and whole-cell recordings demonstrated that astrocytes exhibited a transient Ca(2+)-dependent cell volume increase, which activated glutamate permeable channels. HPLC analysis revealed that glutamate was released in conjunction with other amino acid osmolytes. Our observations indicate that volume-sensitive channel may constitute a previously uncharacterized target for modulation of astrocyte-neuronal interactions. PMID- 16254052 TI - Degradation of the membrane-localized virulence activator TcpP by the YaeL protease in Vibrio cholerae. AB - A common mechanism inhibiting the activity of transcription factors is their sequestration to the membrane until they are needed, at which point they are released from the membrane by proteolysis. Acting in contrast to this inhibition mechanism are virulence regulators of Vibrio cholerae, the ToxR and TcpP proteins, which are localized to the inner membrane of the cell, where they bind promoter DNA and activate gene expression. TcpP is rapidly degraded in the absence of another protein, TcpH. We used a genetic screen to identify regulators of TcpP stability and identified the YaeL membrane-localized zinc metalloprotease as responsible for degrading TcpP in the absence of TcpH. In Escherichia coli, DegS and YaeL cooperate to degrade RseA, an antisigma factor that sequesters sigma(E) to the inner membrane, thereby inhibiting the activity of sigma(E). When yaeL was disrupted in a V. cholerae tcpH mutant, we observed accumulation of a lower molecular weight species of TcpP. This observation is consistent with TcpP being partially degraded in the absence of YaeL. A mutant lacking both DegS and YaeL continued to accumulate the TcpP degradation product, indicating that protease other than DegS is acting before YaeL in degrading TcpP. The YaeL dependent degradation pathway is active in TcpH(+) cells under conditions that are not favorable for virulence gene activation. This work expands the knowledge of YaeL-dependent processing in the bacterial cell and reveals an unexpected layer of virulence gene regulation in V. cholerae. PMID- 16254053 TI - Interplay among coactivator-associated arginine methyltransferase 1, CBP, and CIITA in IFN-gamma-inducible MHC-II gene expression. AB - Class II major histocompatibility (MHC-II) genes are prototype targets of IFN gamma. IFN-gamma activates the expression of the non-DNA-binding master regulator of MHC-II, class II transactivator (CIITA), which is crucial for enhanceosome formation and gene activation. This report shows the importance of the histone methyltransferase, coactivator-associated arginine methyltransferase (CARM1/PRMT4), during IFN-gamma-induced MHC-II gene activation. It also demonstrates the coordinated regulation of CIITA, CARM1, and the acetyltransferase cyclic-AMP response element binding (CREB)-binding protein (CBP) during this process. CARM1 synergizes with CIITA in activating MHC-II transcription and synergy is abrogated when an arginine methyltransferase defective CARM1 mutant is used. Protein-arginine methyltransferase 1 has much less effect on MHC-II transcription. Specific RNA interference reduced CARM1 expression as well as MHC-II expression. The recruitment of CARM1 to the promoter requires endogenous CIITA and results in methylation of histone H3-R17; hence, CIITA is an upstream regulator of histone methylation. Previous work has shown that CARM1 can methylate CBP at three arginine residues. Using wild-type CBP and a mutant of CBP lacking the CARM1-targeted arginine residues (R3A), we show that arginine methylation of CBP is required for IFN-gamma induction of MHC-II. A kinetic analysis shows that CIITA, CARM1, and H3-R17 methylation all precede CBP loading on the MHC-II promoter during IFN-gamma treatment. These results suggest functional and temporal relationships among CIITA, CARM1, and CBP for IFN-gamma induction of MHC-II. PMID- 16254054 TI - Induced conformational changes upon Cd2+ binding at photosynthetic reaction centers. AB - Cd(2+) binding at the bacterial photosynthetic reaction center (bRC) from Rhodobacter sphaeroides is known to inhibit proton transfer (PT) from bulk solvent to the secondary quinone Q(B). To elucidate this mechanism, we calculated the pK(a) for residues along the water channels connecting Q(B) with the stromal side based on the crystal structures of WT-bRC and Cd(2+)-bound bRC. Upon Cd(2+) binding, we observed the release of two protons from His-H126/128 at the Cd(2+) binding site and significant pK(a) shifts for residues along the PT pathways. Remarkably, Asp-L213 near Q(B), which is proposed to play a significant role in PT, resulted in a decrease in pK(a) upon Cd(2+) binding. The direct electrostatic influence of the Cd(2+)-positive charge on these pK(a) shifts was small. Instead, conformational changes of amino acid side chains induced electrostatically by Cd(2+) binding were the main mechanism for these pK(a) shifts. The long-range electrostatic influence over approximately 12 A between Cd(2+) and Asp-L213 is likely to originate from a set of Cd(2+)-induced successive reorientations of side chains (Asp-H124, His-H126, His-H128, Asp-H170, Glu-H173, Asp-M17, and Asp L210), which propagate along the PT pathways as a "domino" effect. PMID- 16254055 TI - Comparison of three ways to determine and deliver pressure during nasal CPAP therapy for obstructive sleep apnoea. AB - BACKGROUND: The simplest method of initiating and maintaining therapeutic continuous positive airways pressure (CPAP) therapy for obstructive sleep apnoea (OSA) has not been established. METHODS: Ninety eight subjects with OSA requiring CPAP treatment (more than 10 dips in oxygen desaturation of >4% per hour of sleep study and Epworth Sleepiness Score (ESS) >9) were randomised prospectively to three different methods of CPAP delivery for 6 months: (1) autotitration pressure throughout; (2) autotitration pressure for 1 week followed by fixed pressure (95th centile) thereafter; and (3) fixed pressure determined by algorithm (based on neck size and dip rate). Patients and investigators were blind to group allocation. One week after initiation the patients were routinely reviewed by sleep nurses. Study assessments took place before starting CPAP treatment and 1 and 6 months after to assess ESS, maintenance of wakefulness test, 24 hour blood pressure, general health (SF-36), and sleep apnoea related quality of life. CPAP internal monitoring data were also collected. RESULTS: There were no significant differences in any of the outcome measures or CPAP monitoring data between the three groups. The 95th centile CPAP pressures delivered in the 6 month and 1 week autotitration groups were higher than in the algorithm group, but the median pressures were lowest in the 6 month autotitration group. CONCLUSIONS: The method of determining CPAP pressure for treatment of moderate to severe OSA makes no significant difference to clinical outcome measures. The autotitration CPAP machine used has no advantage in this setting over simpler methods of pressure determination. PMID- 16254056 TI - Risk factors for multidrug resistant tuberculosis in Europe: a systematic review. AB - BACKGROUND: The resurgence of tuberculosis (TB) in western countries has been attributed to the HIV epidemic, immigration, and drug resistance. Multidrug resistant tuberculosis (MDR-TB) is caused by the transmission of multidrug resistant Mycobacterium tuberculosis strains in new cases, or by the selection of single drug resistant strains induced by previous treatment. The aim of this report is to determine risk factors for MDR-TB in Europe. METHODS: A systematic review was conducted of published reports of risk factors associated with MDR-TB in Europe. Meta-analysis, meta-regression, and sub-grouping were used to pool risk estimates of MDR-TB and to analyse associations with age, sex, immigrant status, HIV status, occurrence year, study design, and area of Europe. RESULTS: Twenty nine papers were eligible for the review from 123 identified in the search. The pooled risk of MDR-TB was 10.23 times higher in previously treated than in never treated cases, with wide heterogeneity between studies. Study design and geographical area were associated with MDR-TB risk estimates in previously treated patients; the risk estimates were higher in cohort studies carried out in western Europe (RR 12.63; 95% CI 8.20 to 19.45) than in eastern Europe (RR 8.53; 95% CI 6.57 to 11.06). National estimates were possible for six countries. MDR-TB cases were more likely to be foreign born (odds ratio (OR) 2.46; 95% CI 1.86 to 3.24), younger than 65 years (OR 2.53; 95% CI 1.74 to 4.83), male (OR 1.38; 95% CI 1.16 to 1.65), and HIV positive (OR 3.52; 95% CI 2.48 to 5.01). CONCLUSIONS: Previous treatment was the strongest determinant of MDR-TB in Europe. Detailed study of the reasons for inadequate treatment could improve control strategies. The risk of MDR-TB in foreign born people needs to be re evaluated, taking into account any previous treatment. PMID- 16254057 TI - Combined donor specific transfusion and anti-CD154 therapy achieves airway allograft tolerance. AB - BACKGROUND: The state of tolerance allows long term graft survival without immunosuppressants. Lung transplantation tolerance has not been consistently achieved in either small or large animal models. METHODS: The mechanisms and effectiveness of a tolerance induction protocol consisting of donor specific transfusion (DST; day 0) and a short course of co-stimulatory blockade (anti CD154 antibody; days -7, -4, 0 and +4) were studied in the mouse heterotopic tracheal transplant model of chronic lung rejection. C57BL/6 mice received BALB/c tracheal grafts (day 0) and were treated with DST alone, anti-CD154 alone, the combination (DST/anti-CD154), or no treatment. No non-specific immunosuppressants were used. RESULTS: DST/anti-CD154 in combination, but neither treatment alone, markedly prolonged the lumen patency and survival (>100 days) of fully histo incompatible allografts (p<0.05 versus control allografts at every time point studied up to 16 weeks) without immunosuppression. This protocol was donor antigen specific as third party grafts (C3H) were promptly rejected. In addition, DST/anti-CD154 did not result in mixed chimerism but induced transplantation tolerance via a peripheral mechanism(s), which included significantly reduced cytotoxic T cell activity (p<0.001) and a significantly increased percentage of CD4+CD25+ cells (p = 0.03). CONCLUSIONS: The DST/anti-CD154 protocol successfully induced and maintained long term, donor specific tolerance in the mouse heterotopic airway graft model of chronic lung rejection. This finding may lead us closer to successful tolerance induction in lung transplantation. PMID- 16254058 TI - Reversing hypoxic cell chemoresistance in vitro using genetic and small molecule approaches targeting hypoxia inducible factor-1. AB - The resistance of hypoxic cells to conventional chemotherapy is well documented. Using both adenovirus-mediated gene delivery and small molecules targeting hypoxia-inducible factor-1 (HIF-1), we evaluated the impact of HIF-1 inhibition on the sensitivity of hypoxic tumor cells to etoposide. The genetic therapy exploited a truncated HIF-1alpha protein that acts as a dominant-negative HIF 1alpha (HIF-1alpha-no-TAD). Its functionality was validated in six human tumor cell lines using HIF-1 reporter assays. An EGFP-fused protein demonstrated that the dominant-negative HIF-1alpha was nucleus-localized and constitutively expressed irrespective of oxygen tension. The small molecules studied were quinocarmycin monocitrate (KW2152), its analog 7-cyanoquinocarcinol (DX-52-1), and topotecan. DX-52-1 and topotecan have been previously established as HIF-1 inhibitors. HT1080 and HCT116 cells were treated with either AdHIF-1alpha-no-TAD or nontoxic concentrations (0.1 microM; or = 0.242), indicating that the quantity of contractile machinery was not affected by removing ovarian hormones. EPR spectroscopy showed that the fraction of strong-binding myosin during contraction was 15% lower in EDL muscles from OVX mice compared with shams [0.277 (SD 0.039) vs. 0.325 (SD 0.020); P = 0.004]. These results indicate that the loss of ovarian hormones has detrimental effects on skeletal muscle force-generating capacities that can be explained by altered actin-myosin interactions. PMID- 16254071 TI - On the modeling of breath-by-breath oxygen uptake kinetics at the onset of high intensity exercises: simulated annealing vs. GRG2 method. AB - Modeling in the time domain, the non-steady-state O2 uptake on-kinetics of high intensity exercises with empirical models is commonly performed with gradient descent-based methods. However, these procedures may impair the confidence of the parameter estimation when the modeling functions are not continuously differentiable and when the estimation corresponds to an ill-posed problem. To cope with these problems, an implementation of simulated annealing (SA) methods was compared with the GRG2 algorithm (a gradient-descent method known for its robustness). Forty simulated Vo2 on-responses were generated to mimic the real time course for transitions from light- to high-intensity exercises, with a signal-to-noise ratio equal to 20 dB. They were modeled twice with a discontinuous double-exponential function using both estimation methods. GRG2 significantly biased two estimated kinetic parameters of the first exponential (the time delay td1 and the time constant tau1) and impaired the precision (i.e., standard deviation) of the baseline A0, td1, and tau1 compared with SA. SA significantly improved the precision of the three parameters of the second exponential (the asymptotic increment A2, the time delay td2, and the time constant tau2). Nevertheless, td2 was significantly biased by both procedures, and the large confidence intervals of the whole second component parameters limit their interpretation. To compare both algorithms on experimental data, 26 subjects each performed two transitions from 80 W to 80% maximal O2 uptake on a cycle ergometer and O2 uptake was measured breath by breath. More than 88% of the kinetic parameter estimations done with the SA algorithm produced the lowest residual sum of squares between the experimental data points and the model. Repeatability coefficients were better with GRG2 for A1 although better with SA for A2 and tau2. Our results demonstrate that the implementation of SA improves significantly the estimation of most of these kinetic parameters, but a large inaccuracy remains in estimating the parameter values of the second exponential. PMID- 16254072 TI - Segment-specific resistivity improves body fluid volume estimates from bioimpedance spectroscopy in hemodialysis patients. AB - Discrepancies in body fluid estimates between segmental bioimpedance spectroscopy (SBIS) and gold-standard methods may be due to the use of a uniform value of tissue resistivity to compute extracellular fluid volume (ECV) and intracellular fluid volume (ICV). Discrepancies may also arise from the exclusion of fluid volumes of hands, feet, neck, and head from measurements due to electrode positions. The aim of this study was to define the specific resistivity of various body segments and to use those values for computation of ECV and ICV along with a correction for unmeasured fluid volumes. Twenty-nine maintenance hemodialysis patients (16 men) underwent body composition analysis including whole body MRI, whole body potassium (40K) content, deuterium, and sodium bromide dilution, and segmental and wrist-to-ankle bioimpedance spectroscopy, all performed on the same day before a hemodialysis. Segment-specific resistivity was determined from segmental fat-free mass (FFM; by MRI), hydration status of FFM (by deuterium and sodium bromide), tissue resistance (by SBIS), and segment length. Segmental FFM was higher and extracellular hydration of FFM was lower in men compared with women. Segment-specific resistivity values for arm, trunk, and leg all differed from the uniform resistivity used in traditional SBIS algorithms. Estimates for whole body ECV, ICV, and total body water from SBIS using segmental instead of uniform resistivity values and after adjustment for unmeasured fluid volumes of the body did not differ significantly from gold standard measures. The uniform tissue resistivity values used in traditional SBIS algorithms result in underestimation of ECV, ICV, and total body water. Use of segmental resistivity values combined with adjustment for body volumes that are neglected by traditional SBIS technique significantly improves estimations of body fluid volume in hemodialysis patients. PMID- 16254073 TI - Effect of muscle fatigue on internal model formation and retention during reaching with the arm. AB - The motor system adapts to novel dynamic environments by forming internal models that predict the muscle forces needed to move skillfully. The goal of this study was to determine how muscle fatigue affects internal model formation during arm movement and whether an internal model acquired while fatigued could be recalled accurately after rest. Twelve subjects adapted to a viscous force field applied by a lightweight robot as they reached to a target. They then reached while being resisted by elastic bands until they could no longer touch the target. This protocol reduced the strength of the muscles used to resist the force field by approximately 20%. The bands were removed, and subjects adapted again to the viscous force field. Their adaptive ability, quantified by the amount and time constant of adaptation, was not significantly impaired following fatigue. The subjects then rested, recovering approximately 70% of their lost force-generation ability. When they reached in the force field again, their prediction of the force field strength was different than in a nonfatigued state. This alteration was consistent with the use of a higher level of effort than normally used to counteract the force field. These results suggest that recovery from fatigue can affect recall of an internal model, even when the fatigue did not substantially affect the motor system's ability to form the model. Recovery from fatigue apparently affects recall because the motor system represents internal models as a mapping between effort and movement and relies on practice to recalibrate this mapping. PMID- 16254074 TI - Gene CATCHR--gene cloning and tagging for Caenorhabditis elegans using yeast homologous recombination: a novel approach for the analysis of gene expression. AB - Expression patterns of gene products provide important insights into gene function. Reporter constructs are frequently used to analyze gene expression in Caenorhabditis elegans, but the sequence context of a given gene is inevitably altered in such constructs. As a result, these transgenes may lack regulatory elements required for proper gene expression. We developed Gene Catchr, a novel method of generating reporter constructs that exploits yeast homologous recombination (YHR) to subclone and tag worm genes while preserving their local sequence context. YHR facilitates the cloning of large genomic regions, allowing the isolation of regulatory sequences in promoters, introns, untranslated regions and flanking DNA. The endogenous regulatory context of a given gene is thus preserved, producing expression patterns that are as accurate as possible. Gene Catchr is flexible: any tag can be inserted at any position without introducing extra sequence. Each step is simple and can be adapted to process multiple genes in parallel. We show that expression patterns derived from Gene Catchr transgenes are consistent with previous reports and also describe novel expression data. Mutant rescue assays demonstrate that Gene Catchr-generated transgenes are functional. Our results validate the use of Gene Catchr as a valuable tool to study spatiotemporal gene expression. PMID- 16254075 TI - Partial 13C isotopic enrichment of nucleoside monophosphates: useful reporters for NMR structural studies. AB - Analysis of the 13C isotopic labeling patterns of nucleoside monophosphates (NMPs) extracted from Escherichia coli grown in a mixture of C-1 and C-2 glucose is presented. By comparing our results to previous observations on amino acids grown in similar media, we have been able to rationalize the labeling pattern based on the well-known biochemistry of nucleotide biosynthesis. Except for a few notable absences of label (C4 in purines and C3' in ribose) and one highly enriched site (C1' in ribose), most carbons are randomly enriched at a low level (an average of 13%). These sparsely labeled NMPs give less complex NMR spectra than their fully isotopically labeled analogs due to the elimination of most 13C 13C scalar couplings. The spectral simplicity is particularly advantageous when working in ordered systems, as illustrated with guanosine diphosphate (GDP) bound to ADP ribosylation factor 1 (ARF1) aligned in a liquid crystalline medium. In this system, the absence of scalar couplings and additional long-range dipolar couplings significantly enhances signal to noise and resolution. PMID- 16254076 TI - Cleavage of dsRNAs hyper-edited by ADARs occurs at preferred editing sites. AB - Long double-stranded RNAs (dsRNAs) may undergo covalent modification (hyper editing) by adenosine deaminases that act on RNA (ADARs), whereby up to 50-60% of adenosine residues are converted to inosine. Previously, we have described a ribonuclease activity in various cell extracts that specifically targets dsRNAs hyper-edited by ADARs. Such a ribonuclease may play an important role in viral defense, or may alternatively be involved in down-regulation of other RNA duplexes. Cleavage of hyper-edited dsRNA occurs within sequences containing multiple IU pairs but not in duplexes that contain either isosteric GU pairs or Watson-Crick base pairs. Here, we describe experiments aimed at further characterizing cleavage of hyper-edited dsRNA. Using various inosine-containing dsRNAs we show that cleavage occurs preferentially at a site containing both IU and UI pairs, and that inclusion of even a single GU pair inhibits cleavage. We also show that cleavage occurs on both strands within a single dsRNA molecule and requires a 2'-OH group. Strikingly, we show that ADAR1, ADAR2 or dADAR all preferentially generate the preferred cleavage site when hyper-editing a long dsRNA. PMID- 16254077 TI - Alternative splicing and nonsense-mediated mRNA decay regulate mammalian ribosomal gene expression. AB - Messenger RNAs containing premature stop codons are generally targeted for degradation through nonsense-mediated mRNA decay (NMD). This mechanism degrades aberrant transcripts derived from mutant genes containing nonsense or frameshift mutations. Wild-type genes also give rise to transcripts targeted by NMD. For example, some wild-type genes give rise to alternatively spliced transcripts that are targeted for decay by NMD. In Caenorhabditis elegans, the ribosomal protein (rp) L12 gene generates a nonsense codon-bearing alternatively spliced transcript that is induced in an autoregulatory manner by the rpL12 protein. By pharmacologically blocking the NMD pathway, we identified alternatively spliced mRNA transcripts derived from the human rpL3 and rpL12 genes that are natural targets of NMD. The deduced protein sequence of these alternatively spliced transcripts suggests that they are unlikely to encode functional ribosomal proteins. Overexpression of rpL3 increased the level of the alternatively spliced rpL3 mRNA and decreased the normally expressed rpL3. This indicates that rpL3 regulates its own production by a negative feedback loop and suggests the possibility that NMD participates in this regulatory loop by degrading the non functional alternatively spliced transcript. PMID- 16254078 TI - Depletion of TDP 43 overrides the need for exonic and intronic splicing enhancers in the human apoA-II gene. AB - Exon 3 of the human apolipoprotein A-II (apoA-II) gene is efficiently included in the mRNA although its acceptor site is significantly weak because of a peculiar (GU)16 tract instead of a canonical polypyrimidine tract within the intron 2/exon 3 junction. Our previous studies demonstrated that the SR proteins ASF/SF2 and SC35 bind specifically an exonic splicing enhancer (ESE) within exon 3 and promote exon 3 splicing. In the present study, we show that the ESE is necessary only in the proper context. In addition, we have characterized two novel sequences in the flanking introns that modulate apoA-II exon 3 splicing. There is a G-rich element in intron 2 that interacts with hnRNPH1 and inhibits exon 3 splicing. The second is a purine rich region in intron 3 that binds SRp40 and SRp55 and promotes exon 3 inclusion in mRNA. We have also found that the (GU) repeats in the apoA-II context bind the splicing factor TDP-43 and interfere with exon 3 definition. Significantly, blocking of TDP-43 expression by small interfering RNA overrides the need for all the other cis-acting elements making exon 3 inclusion constitutive even in the presence of disrupted exonic and intronic enhancers. Altogether, our results suggest that exonic and intronic enhancers have evolved to balance the negative effects of the two silencers located in intron 2 and hence rescue the constitutive exon 3 inclusion in apoA-II mRNA. PMID- 16254079 TI - The ErbB3 binding protein Ebp1 interacts with Sin3A to repress E2F1 and AR mediated transcription. AB - Ectopic expression of ebp1, a member of the PA2G4 family, inhibits the proliferation and induces the differentiation of human breast and prostate cancer cell lines. Ebp1 inhibits transcription of E2F1 and androgen receptor regulated genes such as prostate specific antigen (PSA) through its interactions with histone deacetylases (HDACs). To further understand Ebp1's interactions with other components of the transcriptional repression machinery, we examined the association of Ebp1 with the corepressor Sin3A. Ebp1 interacted with Sin3A both in vitro and in vivo as demonstrated by glutathione S-transferase (GST) pull-down and coimmunoprecipitation analysis. The C-terminal domain of Ebp1, responsible for its ability to repress transcription and arrest cell growth, was necessary and sufficient for binding Sin3A. The C-terminal domain of Sin3A, containing the paired amphipathic domain 4 and the HDAC interacting domain, bound Ebp1. Recombinant Sin3A bound Ebp1 directly, but recombinant HDAC2 failed to bind Ebp1. Chromatin immunoprecipitation (ChIP) and DNA affinity precipitation analysis demonstrated that Ebp1 and Sin3A associate at the PSA and E2F1 promoters. Functionally, Sin3A enhanced the ability of Ebp1 to repress transcription of androgen receptor (AR) and E2F1 regulated genes. These results demonstrate that Ebp1 participates in transcriptional regulation via its interaction with the Sin3 HDAC. PMID- 16254080 TI - Induction of unique structural changes in guanine-rich DNA regions by the triazoloacridone C-1305, a topoisomerase II inhibitor with antitumor activities. AB - We recently reported that the antitumor triazoloacridone, compound C-1305, is a topoisomerase II poison with unusual properties. In this study we characterize the DNA interactions of C-1305 in vitro, in comparison with other topoisomerase II inhibitors. Our results show that C-1305 binds to DNA by intercalation and possesses higher affinity for GC- than AT-DNA as revealed by surface plasmon resonance studies. Chemical probing with DEPC indicated that C-1305 induces structural perturbations in DNA regions with three adjacent guanine residues. Importantly, this effect was highly specific for C-1305 since none of the other 22 DNA interacting drugs tested was able to induce similar structural changes in DNA. Compound C-1305 induced stronger structural changes in guanine triplets at higher pH which suggested that protonation/deprotonation of the drug is important for this drug-specific effect. Molecular modeling analysis predicts that the zwitterionic form of C-1305 intercalates within the guanine triplet, resulting in widening of both DNA grooves and aligning of the triazole ring with the N7 atoms of guanines. Our results show that C-1305 binds to DNA and induces very specific and unusual structural changes in guanine triplets which likely plays an important role in the cytotoxic and antitumor activity of this unique compound. PMID- 16254081 TI - Predicting candidate genomic sequences that correspond to synthetic functional RNA motifs. AB - Riboswitches and RNA interference are important emerging mechanisms found in many organisms to control gene expression. To enhance our understanding of such RNA roles, finding small regulatory motifs in genomes presents a challenge on a wide scale. Many simple functional RNA motifs have been found by in vitro selection experiments, which produce synthetic target-binding aptamers as well as catalytic RNAs, including the hammerhead ribozyme. Motivated by the prediction of Piganeau and Schroeder [(2003) Chem. Biol., 10, 103-104] that synthetic RNAs may have natural counterparts, we develop and apply an efficient computational protocol for identifying aptamer-like motifs in genomes. We define motifs from the sequence and structural information of synthetic aptamers, search for sequences in genomes that will produce motif matches, and then evaluate the structural stability and statistical significance of the potential hits. Our application to aptamers for streptomycin, chloramphenicol, neomycin B and ATP identifies 37 candidate sequences (in coding and non-coding regions) that fold to the target aptamer structures in bacterial and archaeal genomes. Further energetic screening reveals that several candidates exhibit energetic properties and sequence conservation patterns that are characteristic of functional motifs. Besides providing candidates for experimental testing, our computational protocol offers an avenue for expanding natural RNA's functional repertoire. PMID- 16254082 TI - Sources of situational variation in ethnic identity and psychological well-being: a Palm Pilot study of Chinese American students. AB - Experience sampling reports collected randomly six times a day for 1 week examined the association between context, ethnic salience, psychological well being, and stable ethnic centrality and ethnic private regard among Chinese American university students (N = 62). At the level of the situation, ethnic composition, family, and language were associated with higher ethnic salience. Stable ethnic centrality moderated the association between ethnic salience and family where higher centrality was related to stronger ethnic salience when students were with family. At the situation level, ethnic salience was associated with fewer depressive symptoms and increased positive mood. Stable ethnic private regard moderated this association where higher private regard was related to fewer depressive symptoms and higher levels of positive mood when ethnicity was salient. PMID- 16254083 TI - Relationship power and betrayal experience as predictors of reactions to infidelity. AB - Explanations for sexual infidelity have been dominated by an evolutionary psychological theory of jealousy that finds its strongest support in research that employs a forced-choice hypothetical infidelity paradigm wherein participants imagine experiencing infidelity and choose whether sexual or emotional infidelity would be more distressing. Robust gender differences that support evolutionary psychological perspectives are consistently found using this paradigm, but recent work suggests that gender differences may be attenuated among actual infidelity victims. However, no research has used the forced-choice paradigm to compare real and hypothetical infidelity. This study uses this paradigm to compare reactions to imagined dating infidelity to those of infidelity victims. No gender differences are observed in response to the forced choice question among victims of infidelity. Gender differences among participants who imagined infidelity are partially mediated by level of relationship power. Difficulties with the hypothetical forced-choice paradigm and implications for the evolutionary psychological theory of jealousy are discussed. PMID- 16254084 TI - Who I am, who we are, and why: links between emotions and causal attributions for self- and group discrepancies. AB - Discrepancies between people's ought selves and their actual selves and their ideal selves and actual selves predict the emotions that individuals experience. The authors predicted that internal versus external causal attributions for self discrepancies should moderate the relationship between self-discrepancies and emotions, resulting in more refined predictions for both agitation- and dejection related emotions and for two additional types of emotion, namely, anger-related and discontent-related emotions. Results of two studies generally supported the predictions that agitation-related emotions and dejection-related emotions were positively associated with actual-ought discrepancies and actual-ideal discrepancies, respectively, only when causal attributions for the discrepancies were internally based. Anger-related emotions and emotions of discontent were positively associated with actual-ought and actual-ideal discrepancies, respectively, primarily when causal attributions were externally based. Study 2, which addressed group discrepancies and group-based emotions, generally replicated the findings when group identification was high, yielding a more complex model of the link between discrepancies and emotions. PMID- 16254085 TI - Concentrating on beauty: sexual selection and sociospatial memory. AB - In three experiments, location memory for faces was examined using a computer version of the matching game Concentration. Findings suggested that physical attractiveness led to more efficient matching for female faces but not for male faces. Study 3 revealed this interaction despite allowing participants to initially see, attend to, and match the attractive male faces in the first few turns. Analysis of matching errors suggested that, compared to other targets, attractive women were less confusable with one another. Results are discussed in terms of the different functions that attractiveness serves for men and women. PMID- 16254086 TI - Miscommunications surrounding efforts to reach out across group boundaries. AB - Two studies examined the communication of friendship interest within versus across group boundaries in relatively intimate exchanges. The main hypothesis was that so long as reasonable levels of friendship interest were in place, individuals would be more apt to exaggerate the clarity of their overtures (i.e., to exhibit a signal amplification bias) in intergroup as compared to within-group interaction. In line with predictions, this pattern was evident for lower prejudice individuals, who were equally interested in ingroup and outgroup members as potential friends, but not for higher-prejudice individuals, who were relatively disinterested in cross-group friendship and instead tended to underestimate the friendship interest they conveyed to outgroup members. In contrast with the implications of past research centering on trait impressions and impersonal exchanges, both of these effects appeared to be driven by heightened feelings of transparency in intergroup as compared to within-group interaction. PMID- 16254087 TI - Organization and predictive power of general and relationship-specific attachment models: one for all, and all for one? AB - Given recent evidence for multiple attachment models, we examined the organization and predictive power of general and relationship-specific attachment representations in two samples using two distinct measures of attachment models. With regard to associations among relationship-specific models, peer models (romantic partner and friend) and parental models (mother and father) were more similar to each other than to any other models, and anxiety/self-model representations were more consistent across relationships than avoidance/other model representations. With regard to links between general and specific models, romantic and friend models made the strongest and independent contributions to general models, and romantic relationship involvement moderated the importance of romantic models to general models. With regard to differential predictive power of multiple models, general, romantic partner, and mother attachment made unique contributions to well-being indicators; relationship outcomes, however, were only predicted by individuals' corresponding relationship-specific models. Implications for the measurement and conceptualization of adult attachment are discussed. PMID- 16254088 TI - Democracy justifies the means: political group structure moderates the perceived legitimacy of intergroup aggression. AB - The present research tested the hypothesis that the political structure of conflicting groups moderates perceived legitimacy of intergroup aggression. In two experiments, participants read scenarios of fictitious summer camps in which members of one group aggressed members of another group. The political structure of both the perpetrator and the victim groups was described as either egalitarian (defined with democratic decision-making procedures) or hierarchical (authoritarian decision-making procedures). Results of both experiments showed that aggressions perpetrated by members of egalitarian groups at the expense of members of hierarchical groups were evaluated as less illegitimate than aggressions committed in the three remaining conditions. This effect is discussed as a function of the higher social value attributed to democratic groups. PMID- 16254089 TI - Moments in time: metacognition, trust, and outcomes in dyadic negotiations. AB - This research tested the relationships between turning points, cognitive and affective trust, and negotiation outcomes. After completing a simulated negotiation, participants identified turning points from videotape. Turning points were then classified as substantive (interest, offer), characterization (positive, negative), or procedural (positive, negative). Prenegotiation affective trust predicted subsequent turning points, whereas prenegotiation cognitive trust did not, suggesting that different cues influence the two types of trust. Postnegotiation cognitive trust was increased by the occurrence of interest, positive characterization, and positive procedural turning points and decreased by negative characterization turning points. Affective trust was increased by positive procedural turning points. Finally, interest turning points resulted in higher joint outcomes, whereas negative characterization turning points resulted in lower joint outcomes. We conclude that there are two paths to building trust and increasing joint gain, one through insight and one through signaling good faith intentions. PMID- 16254090 TI - The "frozen in time" effect in evaluations of the dead. AB - Two experiments tested the hypothesis that evaluations of the dead are more resistant to change than are evaluations of the living. In Experiment 1, perceivers formed an impression of a target person who performed either a moral or an immoral action and then either died or remained alive. Perceivers were later given new inconsistent information about the target's morality. The results revealed that perceivers' original impressions of the target were significantly less likely to change in response to the inconsistent information when the target was believed to be dead than when she was believed to be alive. Experiment 2 replicated the effect in impressions of real-world targets. The implications of these findings for research on posthumous impression processes are discussed. PMID- 16254091 TI - System-justifying beliefs and psychological well-being: the roles of group status and identity. AB - This research examined the relationship between endorsing system-justifying beliefs and psychological well-being among individuals from ethnic groups that vary in social status. System-justifying beliefs are beliefs that imply that status in society is fair, deserved, and merited; examples of system-justifying beliefs in the United States include the beliefs that hard work pays off and that anyone can get ahead regardless of their group membership. We found that endorsing system-justifying beliefs was negatively related to psychological well being among members of low-status groups who were highly identified with their group but positively related to well-being among members of low-status groups who were not highly identified with their ethnic group. In addition, we found that endorsing system-justifying beliefs was positively related to well-being among members of high-status groups, especially among members of high-status groups who were highly identified with the group. PMID- 16254092 TI - The effects of meritocracy beliefs on women's well-being after first-time gender discrimination. AB - This study examined how meritocracy beliefs may buffer women from the negative psychological effects of an acute situation of gender discrimination. Although some research indirectly suggests that believing that meritocracy exists may increase wellbeing, group consciousness theories suggest that disbelieving that meritocracy exists will enhance psychological adjustment to gender discrimination. Women who reported little past experience with discrimination, and either believed or disbelieved that meritocracy exists, were exposed to either a laboratory situation of discrimination or a nondiscrimination failure (control) condition. Consistent with group consciousness theories, women experiencing discrimination reported greater well-being if they disbelieved that meritocracy exists than if they were believers. In contrast, women in the control condition reported greater wellbeing if they believed that meritocracy exists than if they were disbelievers. Implications for coping with discrimination are discussed. PMID- 16254093 TI - Achievement goals, task performance, and interest: why perceived goal difficulty matters. AB - In field studies, mastery goals, which focus on developing skill, often predict task interest but not actual performance. Performance-approach goals, which focus on outperforming others, instead often predict strong performance but not interest. Two experiments tested the hypothesis that these distinct goal effects trace to goal difficulty perceptions. In each study, participants assigned to a performance-approach goal perceived their goal to be harder, and therefore felt more performance pressure, than those assigned to a mastery goal. Among participants low in dispositional achievement orientation, this experience translated into lower task interest when pursuing the performance-approach goal. However, participants in both studies also performed the activity better when pursuing this goal instead of a standard mastery goal, although this was not mediated by self-reported goal difficulty perceptions. Finally, further demonstrating the role of goal difficulty, a mastery goal manipulated to appear more difficult than a standard mastery goal produced effects matching the performance-approach goal. PMID- 16254096 TI - The molecular pathogenetic role of cell adhesion in endocrine neoplasia. AB - It is becoming increasingly evident that cell adhesion is an important determinant of organised growth and the maintenance of architectural integrity. Indeed, reduced adhesiveness between cells and with the extracellular matrix is a hallmark of neoplastic growth. In neuroendocrine tissues, neural cell adhesion molecule is implicated in modulating cell growth, migration, and differentiation. This review will focus on the molecular pathways involving key growth factor receptors that govern normal adhesive forces. The extent to which disruption of these adhesive forces contributes to the tumorigenic process in neuroendocrine tissues will be highlighted. Validation of the functional relevance of these adhesive pathways will be discussed in light of targeted pharmacotherapeutic studies that are unmasking novel approaches to the treatment of neuroendocrine tumours. PMID- 16254099 TI - Terminology for the diagnosis of colitis. PMID- 16254097 TI - The role of enterovirus in chronic fatigue syndrome. AB - Two and a half decades after coining of the term chronic fatigue syndrome (CFS), the diagnosis of this illness is still symptom based and the aetiology remains elusive. Enteroviruses are well known causes of acute respiratory and gastrointestinal infections, with tropism for the central nervous system, muscles, and heart. Initial reports of chronic enteroviral infections causing debilitating symptoms in patients with CFS were met with skeptism, and had been largely forgotten for the past decade. Observations from in vitro experiments and from animal models clearly established a state of chronic persistence through the formation of double stranded RNA, similar to findings reported in muscle biopsies of patients with CFS. Recent evidence not only confirmed the earlier studies, but also clarified the pathogenic role of viral RNA through antiviral treatment. This review summarises the available experimental and clinical evidence that supports the role of enterovirus in chronic fatigue syndrome. PMID- 16254100 TI - Pathology parameters and adjuvant tamoxifen response in a randomised premenopausal breast cancer trial. AB - BACKGROUND: Subgroups of breast cancer that have an impaired response to endocrine treatment, despite hormone receptor positivity, are still poorly defined. Breast cancer can be subdivided according to standard pathological parameters including histological type, grade, and assessment of proliferation. These parameters are the net result of combinations of genetic alterations effecting tumour behaviour and could potentially reflect subtypes that respond differently to endocrine treatment. AIMS: To investigate the usefulness of these parameters as predictors of the response to tamoxifen in premenopausal women with breast cancer. MATERIALS/METHODS: Clinically established pathological parameters were assessed and related to the tamoxifen response in 500 available tumour specimens from 564 premenopausal patients with breast cancer randomised to either two years of tamoxifen or no treatment with 14 years of follow up. Proliferation was further evaluated by immunohistochemical Ki-67 expression. RESULTS: Oestrogen receptor positive ductal carcinomas responded as expected to tamoxifen, whereas the difference in recurrence free survival between control and tamoxifen treated patients was less apparent in the relatively few lobular carcinomas. For histological grade, there was no obvious difference in treatment response between the groups. The relation between proliferation and tamoxifen response seemed to be more complex, with a clear response in tumours with high and low proliferation, whereas tumours with intermediate proliferation defined by Ki-67 responded more poorly. CONCLUSIONS: Clinically established pathology parameters seem to mirror the endocrine treatment response and could potentially be valuable in future treatment decisions for patients with breast cancer. PMID- 16254101 TI - Evidence based criteria for the histopathological diagnosis of toxoplasmic lymphadenopathy. AB - AIMS: To formulate evidence based histopathological criteria for the diagnosis of acquired toxoplasmic lymphadenitis, in an area of high tuberculosis prevalence. METHODS: Multiple histopathological parameters were assessed in a consecutive sample of biopsies from 68 patients presenting with lymphadenopathy with a duration of less than six months. Serum IgM enzyme linked immunosorbent assay was used as the standard reference test for the diagnosis of toxoplasmic lymphadenitis. The sensitivity, specificity, and likelihood ratios of various histological parameters were estimated. RESULTS: The presence of microgranulomas (p < 0.0001), paracortical widening (p = 0.006), paracortical hyperplasia (p = 0.02), monocytoid B cells in sinuses (p = 0.007), lower than grade 2 macrogranuloma (p = 0.002), and the absence of giant cells (p = 0.05) were found to discriminate between IgM positive cases and IgM negative controls. Using a composite criterion-(1) presence of microgranulomas, (2) lower than grade 2 macrogranuloma, (3) absence of giant cells, and (4) follicular hyperplasia toxoplasmic lymphadenitis can be diagnosed with a high degree of sensitivity (100%), specificity (96.6%), and positive likelihood ratio (29). CONCLUSION: Toxoplasma lymphadenitis can be diagnosed with a high degree of confidence using the specific histopathological criteria identified here. PMID- 16254102 TI - Role of mesangial fibrinogen deposition in the pathogenesis of crescentic Henoch Schonlein nephritis in children. AB - AIMS: To clarify the role of mesangial fibrinogen deposition in crescentic Henoch Schonlein nephritis (HSN). METHODS: A retrospective analysis of 21 children with HSN treated with immunosuppressants. Serial renal biopsies were performed before and after treatment. They were divided into two groups according to the immunofluorescent course of fibrinogen deposition: group I (n = 9), no or decreased deposition; group II (n = 12), persistent or increased deposition. RESULTS: There were no differences between the two groups in renal manifestations or laboratory and histological findings at presentation. However, the activity index after immunosuppressive treatment was significantly decreased in group I (mean, 7.9 (SEM, 0.7) v 2.9 (0.4); p = 0.008) and unchanged in group II (mean, 6.8 (SEM, 0.3) v 6.0 (2.1)). The chronicity index was unchanged in group I, but increased in group II (mean, 0.8 (SEM, 0.3) v 1.8 (0.3); p = 0.02). Univariate analysis revealed that the only factor significantly related to persistent or increased fibrinogen deposition was age more than 9 years (p = 0.03). Furthermore, the intensity of fibrinogen deposition at the second biopsy correlated positively with the age at onset (R2= 0.306; p = 0.009) and changes in the percentage of crescents (post-treatment crescents (%) minus pretreatment crescents (%)) correlated positively with the intensity of fibrinogen deposition at the second biopsy (R2= 0.193; p = 0.046). CONCLUSIONS: This study indicates that fibrinogen deposition has an important role to play in renal injury of crescentic HSN and reflects persistent severe histological activity. PMID- 16254103 TI - Immunohistochemical analysis of desmoid tumours. AB - BACKGROUND/AIMS: Although the standard treatment for desmoid tumours is complete surgical resection with wide margins, the optimal adjuvant treatment for recurrent or inoperable disease is unclear, often being based on sporadic immunohistochemical reports with a low number of cases. Therefore, a large immunohistochemical study was performed, to provide a theoretical basis for adjuvant treatment regimens. METHODS: One hundred and sixteen tissue samples from 80 patients (49 female, 31 male; mean age, 34 years; range, 0-83) with desmoid tumours (46 extra-abdominal, 21 abdominal, 13 intra-abdominal) were tested for oestrogen receptors alpha and beta, progesterone and androgen receptors, and somatostatin, in addition to HER2, cathepsin D, Ki-67, and c-KIT by immunohistochemistry. RESULTS: All samples were negative for oestrogen receptor alpha, HER2, and the progesterone receptor. Positive staining for the androgen receptor was found in six extra-abdominal cases. Staining for oestrogen receptor beta was positive in four extra-abdominal, two abdominal, and one intra-abdominal case. Staining for somatostatin was positive in six extra-abdominal, two abdominal, and one intra-abdominal case, and staining for cathepsin D was positive in all cases. Positive staining for Ki-67 was found in 14 extra abdominal, three abdominal, and three intra-abdominal cases. C-KIT was detectable in one abdominal case only. CONCLUSIONS: The data from this immunohistochemical study show that the published effects of antioestrogens and imatinib mesylate in the treatment of aggressive fibromatoses may not be attributable to oestrogen receptor alpha or c-KIT expression. PMID- 16254104 TI - Loss of BCL-2 in the progression of oral cancer is not attributable to mutations. AB - BACKGROUND: BCL-2 and BAX are important in the regulation of apoptosis. There have been reports of loss of BCL-2 in basal cells of oral epithelial dysplasia (OED) and in oral squamous cell carcinoma (OSCC), and suppression of BAX in poorly differentiated OSCC. AIM: To investigate whether loss of BCL-2 in OED and OSCC, and of BAX in poorly differentiated OSCC could be attributed to BCL-2 and BAX mutations. METHODS: Immunohistochemistry and in situ hybridisation were used to confirm BCL-2 and BAX expression. DNA was extracted from archival samples of OED (n = 22) and OSCC (n = 28). The connective tissue part from each section was collected separately and used as the normal reference. RESULTS: No mutations were detected in BCL-2 or BAX that could explain their aberrant expression at the mRNA and protein levels in OED and OSCC. The reported A/G polymorphism at codon 7 of BCL-2 was detected in 18 of 50 samples and a novel C/T polymorphism at codon 100 was detected in three of 50 samples. CONCLUSIONS: No mutations were found that could explain loss of BCL-2 in oral dysplasia and carcinoma. An unreported C/T polymorphism in BCL-2 was detected. Downregulation of BCL-2 in OED and OSCC may be the result of transcriptional regulation. PMID- 16254105 TI - Activation of the extracellular signal regulated kinase (ERK) pathway in human melanoma. AB - BACKGROUND: Several studies suggest that melanoma may be resistant to treatment because of resistance to apoptosis and that this may be the result of activation of the extracellular signal regulated kinase (ERK1/2) pathway. AIMS: To test this hypothesis by examining the expression of ERK1/2 and its activated form in histological sections of melanoma and its relation to known prognostic features of the disease. MATERIALS/METHODS: Immunohistochemistry with antibodies to ERK1/2 and phosphorylated ERK (p-ERK) was performed on formalin fixed sections from 42 primary melanomas, 38 metastases, and 20 naevi. Fourteen of the primary melanomas were in the radial and 28 in the vertical growth phase. RESULTS: ERK1/2 was widely expressed (100%) in all the (pigmented) lesions studied. p-ERK1/2 expression was much lower in compound (32.4%) and dysplastic (54.5%) naevi than in primary melanoma (nodular 78.8%, superficial spreading 67%) and subcutaneous metastases (76.3%). p-ERK expression was much lower in lymph node metastases (48.5%), suggesting that the microenvironment may influence the activation of ERK. There was a (non-significant) trend for p-ERK expression to be higher in thick (>1.0 mm) versus thin (< or =1.0 mm) melanoma (p = 0.23). There was a trend for overall survival to be related to p-ERK expression in patients with melanoma over 1 mm in thickness. CONCLUSIONS: Expression of activated ERK1/2 in melanocytic lesions appears to be related to malignant potential so that activation of ERK1/2 may be important in melanoma progression. These results provide important histological support for the proposal that inhibition of this signalling pathway may be useful in treatment of melanoma. PMID- 16254106 TI - Tumour proliferation, angiogenesis, and ploidy status in human colon cancer. AB - AIMS: Tumour angiogenesis is essential for carcinogenesis and facilitates the process of tumour development and metastasis. Vascular endothelial growth factor (VEGF) is a well characterised angiogenetic factor and is known to play a crucial role in new vessel development. To gain further insight into the effects of microvessel density and VEGF expression in colon cancer, their relation with tumour proliferation, ploidy status, and p53 expression was investigated in colon cancer. METHODS: Tissue samples of 50 archived colon cancers were analysed by immunohistochemistry for VEGF, p53, and the endothelial cell marker, von Willebrand factor (VWF), using specific antibodies. The same samples were re-cut for flow cytometric studies to obtain S phase fraction (SPF) and ploidy status. RESULTS: A positive significant correlation was found between SPF and angiogenesis. The median microvessel count in high SPF tumours was significantly higher than in low SPF ones. No association was found between VEGF expression and SPF. A positive correlation was found between ploidy status and p53 expression and microvessel count. Furthermore, a positive correlation was established between DNA ploidy, VEGF expression, and microvessel count. CONCLUSION: This study provides evidence that in colon cancer, tumour growth may be stimulated by vascular supply, and the lack of a correlation between tumour cell proliferation and VEGF expression indicates that these two parameters may be regulated by separate mechanisms. Furthermore, the positive correlation between microvessel density, VEGF expression, and ploidy status provides more evidence that genetic alterations are involved in tumour angiogenesis. PMID- 16254107 TI - Aberrant expression of DeltaNp73 in benign and malignant tumours of the prostate: correlation with Gleason score. AB - BACKGROUND: The p73 gene is a p53 homologue that induces apoptosis and inhibits cell proliferation. N-terminal truncated isoforms of p73 (DeltaNp73) act as dominant-negative inhibitors of wild-type p53 and TAp73 and result in tumour growth in nude mice. AIMS: To detect DeltaNp73 expression in 24 benign prostatic hyperplasia samples, 33 prostate carcinomas, and five normal samples and to evaluate the relation between DeltaNp73, TAp73 concentrations, and the clinicopathological characteristics of patients with prostate cancer. METHODS: TAp73 was determined by real time polymerase chain reaction (PCR); DeltaNp73 and DeltaN'p73 were assessed using reverse transcription PCR. western blotting was used to analyse protein expression. p53 mutation was determined by immunohistochemistry. RESULTS: A significant increase of DeltaNp73 was seen in 20 of 33 carcinomas and 17 of 24 benign prostate hyperplasia tissues, but in none of the normal samples. None of the specimens expressed DeltaN'p73. No significant relation was found between TAp73 expression and clinical parameters. The incidence of positive expression of DeltaNp73 correlated with the Gleason score in prostate carcinomas. Cancer samples with wild-type p53 had significantly higher expression of DeltaNp73 than p53 mutant cancers. CONCLUSION: These data suggest a potential role for DeltaNp73 in prostate cancer progression. PMID- 16254108 TI - PCR based identification and discrimination of agents of mucormycosis and aspergillosis in paraffin wax embedded tissue. AB - BACKGROUND: Invasive fungal infections are often diagnosed by histopathology without identification of the causative fungi, which show significantly different antifungal susceptibilities. AIMS: To establish and evaluate a system of two seminested polymerase chain reaction (PCR) assays to identify and discriminate between agents of aspergillosis and mucormycosis in paraffin wax embedded tissue samples. METHODS: DNA of 52 blinded samples from five different centres was extracted and used as a template in two PCR assays targeting the mitochondrial aspergillosis DNA and the 18S ribosomal DNA of zygomycetes. RESULTS: Specific fungal DNA was identified in 27 of 44 samples in accordance with a histopathological diagnosis of zygomycosis or aspergillosis, respectively. Aspergillus fumigatus DNA was amplified from one specimen of zygomycosis (diagnosed by histopathology). In four of 16 PCR negative samples no human DNA was amplified, possibly as a result of the destruction of DNA before paraffin wax embedding. In addition, eight samples from clinically suspected fungal infections (without histopathological proof) were examined. The two PCR assays detected a concomitant infection with Absidia corymbifera and A fumigatus in one, and infections with Rhizopus arrhizus and A fumigatus in another two cases. CONCLUSIONS: The two seminested PCR assays described here can support a histopathological diagnosis of mucormycosis or aspergillosis, and can identify the infective agent, thereby optimising antifungal treatment. PMID- 16254109 TI - CD44s is useful in the differentiation of benign and malignant papillary lesions of the breast. AB - BACKGROUND/AIMS: CD44s, the standard form of CD44, has been shown to be downregulated during malignant transformation of breast cancers. It has also been reported recently to be a useful marker in differentiating between benign and malignant papillary lesions of the breast, with high expression in the former. CD44s expression in benign and malignant papillary lesions was evaluated. METHODS: CD44s expression was assessed by immunohistochemistry in 101 benign papillomas and 59 papillary carcinomas (seven invasive papillary carcinomas, 41 papillary ductal carcinomas in situ, and 11 ductal carcinomas involving papillomas). RESULTS: Patients' age and tumour size were significantly different between the papilloma and papillary carcinoma groups (p < 0.0001). CD44s showed positive staining in 45 papillomas (45%) and five papillary carcinomas (8%), and the difference was significant (p < 0.0001). The myoepithelial cells, when present, were also positive for CD44s in both groups, with no observable differences. Using CD44s positive staining to differentiate between benign and malignant papillary lesions gives a sensitivity, specificity, and accuracy of 45%, 92%, and 62%, respectively. CONCLUSIONS: CD44s may be useful as an adjunct in the evaluation of morphologically problematic cases of papillary lesion of the breast. PMID- 16254110 TI - Atrophic gastritis in young children and adolescents. AB - BACKGROUND: Helicobacter pylori associated gastric cancer arises via a multistage process, with atrophic gastritis being the precursor lesion. Helicobacter pylori is typically acquired in childhood, yet little is known of the prevalence of atrophic gastritis in childhood. AIM: To study atrophic gastritis among children from countries with high gastric cancer incidence. METHODS: Sections from topographically mapped gastric biopsy specimens from children undergoing clinically indicated endoscopy in Korea and Colombia were evaluated using visual analogue scales. Atrophy was defined as loss of normal glandular components, including replacement with fibrosis, intestinal metaplasia (IM), and/or pseudopyloric metaplasia of the corpus (identified by the presence of pepsinogen I in mucosa that was topographically corpus but phenotypically antrum). RESULTS: One hundred and seventy three children, 58 from Korea (median age, 14 years) and 115 from Colombia (median age, 13 years), were studied. Helicobacter pylori was present in 85% of Colombian children versus 17% of Korean children (p<0.01). Atrophic mucosa near the antrum-corpus border was present in 16% of children, primarily as pseudopyloric metaplasia (31%, IM; 63%, pseudopyloric metaplasia; 6%, both). The median age of children with corpus atrophy was 15 (range, 7-17) years. CONCLUSION: Gastric atrophy occurs in H pylori infected children living in countries with high gastric cancer incidence. Identification and characterisation of the natural history of H pylori gastritis requires targeted biopsies to include the lesser and greater curve of the corpus, starting just proximal to the anatomical antrum-corpus junction, in addition to biopsies targeting the antrum and cardia. PMID- 16254111 TI - Interobserver reproducibility of histological features in cutaneous malignant melanoma. AB - AIMS: To assess the interobserver reproducibility of certain histological features proposed for the diagnosis of melanoma. METHODS: In a series of melanomas, 13 histological parameters were analysed: dimension > 6 mm, asymmetry, poor circumscription, irregular confluent nests, single melanocytes predominating, absence of maturation, suprabasal melanocytes, asymmetrical melanin, melanin in deep cells, cytological atypia, mitoses, dermal lymphocytic infiltrate, and necrosis. RESULTS: The agreement (reproducibility) between the nine observers was excellent (kappa > 0.75) for 10 of the 13 examined features (dimension > 6 mm, poor circumscription, irregular confluent nests, single melanocytes predominating, absence of maturation, suprabasal melanocytes, asymmetrical melanin, melanin in deep cells, mitoses, and necrosis). The agreement for asymmetry was very close to excellence (kappa = 0.74), and that for cytological atypia (kappa = 0.65) and dermal lymphocytic infiltrate (kappa = 0.47) was slightly lower, but in the fair to good agreement range. The kappa values obtained by comparison with the majority diagnosis were generally high (> or = 0.85); the mean value of kappa was lower (0.70) for only one parameter (dermal lymphocytic infiltrate). CONCLUSIONS: The parameters investigated showed an overall good reproducibility. PMID- 16254112 TI - Prognostic value of activated Akt expression in oral squamous cell carcinoma. AB - BACKGROUND: Akt is a serine/threonine kinase that plays an important role in tumorigenesis and influences prognosis in several cancers. However, its importance in oral squamous cell carcinomas (OSCC) has not been elucidated. AIM: To investigate the association between the expression of activated Akt, clinicopathological factors, and E-cadherin, PCNA (proliferating cell nuclear antigen), and VEGF (vascular endothelial growth factor) expression to verify the validity of Akt as a prognostic factor in OSCC. METHODS: Phosphorylated Akt (p Akt), E-cadherin, PCNA, and VEGF expression were assessed immunohistochemically in 84 OSCCs. The results were analysed in relation to clinicopathological factors. RESULTS: p-Akt was expressed in 29 cases. It was significantly correlated with lymph node metastasis, TNM stage, and E-cadherin expression. Univariate analysis showed that p-Akt expression, E-cadherin expression, PCNA expression, differentiation, tumour size, lymph node metastasis, TNM stage, and recurrence correlated with prognosis. Multivariate analysis showed that p-Akt expression is an independent prognostic factor in patients with OSCC. CONCLUSIONS: This study revealed that Akt activation is a significant prognostic indicator for OSCC and is correlated with E-cadherin expression. The inhibition of Akt is a possible molecular approach to the treatment of OSCC. PMID- 16254114 TI - Alveolar adenoma of the lung: a clinicopathological description of a case of this very unusual tumour. AB - Alveolar adenomas are extremely rare, and are probably benign lung tumours of unknown histogenesis. This report describes a case of alveolar adenoma in a 43 year old white man, who presented with pleuritic chest pain. A chest x ray and computerised tomography scan demonstrated a solitary left lower lobe lung nodule. Although a positron emission tomography scan seemed to document the benign nature of the lesion, a thoracoscopic wedge resection was performed to alleviate the symptoms and verify the diagnosis. Histologically, the lesion was well demarcated, dominated by large and small cysts with no normal lung parenchyma. The interstitial cellular component consisted of both epithelioid and vaguely spindle shaped cells. The cystic cell linings were mostly indistinct, although areas of cuboidal epithelial cells were seen. Multiple histochemical and immunohistochemical tests were performed. There were no histological signs of malignancy and the patient is doing well one and a half years postoperatively. PMID- 16254113 TI - Grading melanocytic dysplasia in paraffin wax embedded tissue by the nucleic acid index. AB - BACKGROUND: Although nucleic acid derangements are the hallmark of melanocytic dysplasia, the gold standard for its diagnosis remains the microscopic evaluation of haematoxylin and eosin stained slides. However, light microscopy is subjective and crucial genomic changes do not always show as changes in histology. AIMS: To introduce the nucleic acid index (NAI) as a means of analysing nucleic acid derangements in histological sections at the level of the individual cell and within the context of its microenvironment. METHODS: Confocal laser scanning microscopy was performed on melanocytic lesions stained with acridine orange (AO), a fluorescent stain for DNA and RNA. The NAI, calculated by measuring the fluorescence intensities of AO in nuclei relative to the surrounding cytoplasm, reflects the concentration of DNA relative to RNA. RESULTS: When applied to benign naevi, dysplastic naevi, and melanoma, a very strong significant association was seen between lower NAI and malignant potential (p < 0.0001). Strong inverse correlations were found between NAI and both mitotic index and Breslow thickness. Interestingly, the NAI for dysplastic naevi is between that of melanoma and most benign naevi, consistent with their intermediate biological behaviour and histological appearance. CONCLUSION: By providing a quantitative measure for melanocytic neoplasia, the NAI may improve the diagnosis of melanocytic lesions and the selection of treatment. PMID- 16254116 TI - Mucinous metaplasia of the vulva in a case of lichen sclerosus. A case report. AB - Mucinous metaplasia of the genital area is a rare condition characterised by the emergence of mucin containing cells in stratified squamous epithelium. This report describes a unique case of benign mucinous metaplasia of the vulva associated with lichen sclerosus in a 60 year old woman. Histopathology revealed cervical type metaplasia with otherwise typical lichen sclerosus. This report discusses the pathogenesis and differential diagnosis of mucinous epithelium. PMID- 16254115 TI - Rat bite fever caused by Streptobacillus moniliformis in a child: human infection and rat carriage diagnosed by PCR. AB - A child owning pet rats developed an eruptive fever with blisters, polyarthritis, and spectacular desquamation of the hands. Streptobacillus moniliformis was identified after culture of the child's blister fluid and was detected in rat samples by molecular methods. Such detection in the pet of a human victim of rat bite fever has not been reported previously. PMID- 16254117 TI - An unusual case of hyponatraemia in diabetic ketoacidosis. AB - This report outlines a case of diabetic ketoacidosis associated hyponatraemia in an 18 year old woman with type 1 diabetes who presented to the accident and emergency department and was quickly admitted to the intensive treatment unit. Causes of hyponatraemia include sodium depletion, pseudohyponatraemia, and extracellular hypertonicity. Hypertonicity secondary to hyperglycaemia is thought to be the major cause of hyponatraemia in diabetic ketoacidosis. Indirect and direct sodium measurements were performed until the glucose concentration stabilised. The large difference between the presenting sodium concentrations is consistent with pseudohyponatraemia. However, the causes of pseudohyponatraemia (large increases in total protein, triglyceride, and cholesterol concentrations) were excluded. Analytical error should always be considered when the laboratory results do not agree with the clinical picture. Sometimes, however, even after excluding all known effects, the cause may remain unexplained, as in this case. PMID- 16254119 TI - Solitary ovarian splenosis. AB - This report describes the case of an asymptomatic, solitary splenotic nodule in the right ovary, revealed incidentally at histopathological examination. In gynaecological patients, splenosis most often presents as multiple pelvic nodules mimicking endometriosis. Two cases of ovarian splenosis accompanying pelvic and serosal splenotic nodules have been reported previously. However, this is the first documented case of solitary intraovarian splenosis. PMID- 16254118 TI - Serous adenocarcinoma of the sigmoid mesentery arising in cystic endosalpingiosis. AB - This case report describes a Mullerian serous adenocarcinoma arising within a multoloculated cyst lined by ciliated serous-type epithelium located in the sigmoid mesentery. Twenty years previously the patient underwent a hysterectomy, bilateral salpingo-oophorectomy, and omentectomy. The ovaries contained bilateral serous cystadenofibromas, and multiple cysts lined by ciliated serous-type epithelium were present in the omentum. The resection specimen 20 years later contained a 14 cm multiloculated cyst located in the sigmoid mesentery. This was lined largely by benign ciliated serous-type epithelium but a focus of well differentiated serous adenocarcinoma projected into the lumen. Two further peritoneal cysts were present, both of which were lined by ciliated serous-type epithelium. There was a coincidental renal cell carcinoma. This is a unique case of multiple omental, peritoneal, and retroperitoneal cysts (classified as cystic endosalpingiosis), one of which developed a focus of serous adenocarcinoma. Although rarely serous adenocarcinomas, similar to those occurring within the ovary, arise in the retroperitoneum, this is the first reported occurrence in association with a pre-existing benign lesion. PMID- 16254120 TI - Solitary epithelial cells in B cell gastric MALT lymphoma. AB - BACKGROUND: Gastric mucosa associated lymphoid tissue (MALT) lymphoma is a low grade B cell lymphoma histologically characterised by neoplastic B cells surrounding follicles in a marginal zone pattern and selectively infiltrating epithelium to form characteristic lymphoepithelial lesions. AIMS: To identify solitary epithelial cells in gastric MALT lymphoma and investigate their nature. METHODS: Anonymised endoscopic biopsies from eight B cell gastric MALT lymphomas and 10 control biopsies from chronic atrophic gastritis were selected. The numbers of solitary cytokeratin positive epithelial cells were assessed both semiquantitatively and quantitatively in immunostained sections. Chromogranin A expression was studied in sections consecutive to those stained for cytokeratin. RESULTS: Statistical analysis of the quantitative data confirmed that solitary epithelial cells were significantly more common in the lymphomas. The study of consecutive sections showed that the single cells express chromogranin A. CONCLUSIONS: The presence of solitary, cytokeratin positive epithelial cells within the neoplastic infiltrate is a characteristic feature of gastric B cell lymphoma. These solitary epithelial cells are of neuroendocrine origin. PMID- 16254121 TI - Rare allelic imbalances, but no mutations of the PRDX1 gene in human hepatocellular carcinomas. AB - Allelic losses on chromosome 1p are frequent in hepatocellular carcinoma (HCC), suggesting the presence of a tumour suppressor gene in this region. The gene for peroxiredoxin 1 (PRDX1), an antioxidant enzyme, has been mapped to 1p34.1. Mice lacking PRDX1 develop HCC with high frequency. Because oxidative stress has been implicated in the pathogenesis of HCC, this study was designed to determine whether the PRDX1 gene is mutated in human HCC using loss of heterozygosity (LOH) analysis, polymerase chain reaction/denaturing gradient gel electrophoresis, and DNA sequencing. LOH of at least one of four microsatellite markers within 0.8 Mb of the PRDX1 gene was seen in three of 34 informative HCCs, but no mutations or polymorphisms in the translated exons 2-6 of the PRDX1 gene were found. These results suggest that genetic alterations of the PRDX1 locus are rare events in human HCC, indicating that other genes on chromosome 1p contribute to liver carcinogenesis. PMID- 16254122 TI - Cystic atrioventricular node tumour: not a mesothelioma. PMID- 16254123 TI - Role of nitrosyl factors in the hindlimb vasodilation elicited by baroreceptor afferent nerve stimulation. AB - This study determined whether electrical stimulation (ES) of the baroreceptor afferent fibers in the aortic depressor nerve (ADN) produces hindlimb vasodilation in pentobarbital-anesthetized rats via the release of nitric oxide (NO)-containing (nitrosyl) factors from NO synthase-positive lumbar sympathetic nerve terminals. ES of the ADN (1-10 Hz for 15 s) produced frequency-dependent reductions in mean arterial blood pressure (MAP) and mesenteric and hindlimb vascular resistance (MR and HLR, respectively). The falls in resistance were substantially smaller in hindlimb beds in which the ipsilateral lumbar sympathetic chain had been transected 7-10 days previously. The maximal falls in MR and hindquarter vascular resistance (HQR) produced by 1- to 10-Hz ES of the ADN were unaffected by the specific inhibitor of neuronal NO synthase 7 nitroindazole (7-NI, 45 mg/kg iv). However, the total falls in HQR (mmHg.kHz( 1).s) produced by these stimuli were significantly diminished by 7-NI, whereas the total falls in MR were not affected. Four successive episodes of 10-Hz ES produced equivalent reductions in MAP, MR, and HQR. The peak changes in these parameters were not affected by 7-NI. However, the total falls in HQR progressively diminished with each successive stimulus, whereas the total falls in MR remained unchanged. These results provide evidence that the hindlimb vasodilation produced by ES of baroreceptor afferents within the ADN may involve the activation of postganglionic lumbar sympathetic vasodilator fibers, which release newly synthesized and preformed nitrosyl factors. PMID- 16254125 TI - Sprint performance-duration relationships are set by the fractional duration of external force application. AB - We hypothesized that the maximum mechanical power outputs that can be maintained during all-out sprint cycling efforts lasting from a few seconds to several minutes can be accurately estimated from a single exponential time constant (k(cycle)) and two measurements on individual cyclists: the peak 3-s power output (P(mech max)) and the maximum mechanical power output that can be supported aerobically (P(aer)). Tests were conducted on seven subjects, four males and three females, on a stationary cycle ergometer at a pedal frequency of 100 rpm. Peak mechanical power output (P(mech max)) was the highest mean power output attained during a 3-s burst; the maximum power output supported aerobically (P(aer)) was determined from rates of oxygen uptake measured during a progressive, discontinuous cycling test to failure. Individual power output duration relationships were determined from 13 to 16 all-out constant load sprints lasting from 5 to 350 s. In accordance with the above hypothesis, the power outputs measured during all-out sprinting efforts were estimated to within an average of 34 W or 6.6% from P(mech max), P(aer), and a single exponential constant (k(cycle) = 0.026 s(-1)) across a sixfold range of power outputs and a 70-fold range of sprint trial durations (R2 = 0.96 vs. identity, n = 105; range: 180 to 1,136 W). Duration-dependent decrements in sprint cycling power outputs were two times greater than those previously identified for sprint running speed (k(run) = 0.013 s(-1)). When related to the respective times of pedal and ground force application rather than total sprint time, decrements in sprint cycling and running performance followed the same time course (k = 0.054 s(-1)). We conclude that the duration-dependent decrements in sprinting performance are set by the fractional duration of the relevant muscular contractions. PMID- 16254124 TI - Estradiol improves cardiac and hepatic function after trauma-hemorrhage: role of enhanced heat shock protein expression. AB - Although studies indicate that 17beta-estradiol administration after trauma hemorrhage (T-H) improves cardiac and hepatic functions, the underlying mechanisms remain unclear. Because the induction of heat shock proteins (HSPs) can protect cardiac and hepatic functions, we hypothesized that these proteins contribute to the salutary effects of estradiol after T-H. To test this hypothesis, male Sprague-Dawley rats ( approximately 300 g) underwent laparotomy and hemorrhagic shock (35-40 mmHg for approximately 90 min) followed by resuscitation with four times the shed blood volume in the form of Ringer lactate. 17beta-estradiol (1 mg/kg body wt) was administered at the end of the resuscitation. Five hours after T-H and resuscitation there was a significant decrease in cardiac output, positive and negative maximal rate of left ventricular pressure. Liver function as determined by bile production and indocyanine green clearance was also compromised after T-H and resuscitation. This was accompanied by an increase in plasma alanine aminotransferase (ALT) levels and liver perfusate lactic dehydrogenase levels. Furthermore, circulating levels of TNF-alpha, IL-6, and IL-10 were also increased. In addition to decreased cardiac and hepatic function, there was an increase in cardiac HSP32 expression and a reduction in HSP60 expression after T-H. In the liver, HSP32 and HSP70 were increased after T-H. There was no change in heart HSP70 and liver HSP60 after T-H and resuscitation. Estradiol administration at the end of T-H and resuscitation increased heart/liver HSPs expression, ameliorated the impairment of heart/liver functions, and significantly prevented the increase in plasma levels of ALT, TNF-alpha, and IL-6. The ability of estradiol to induce HSPs expression in the heart and the liver suggests that HSPs, in part, mediate the salutary effects of 17beta-estradiol on organ functions after T-H. PMID- 16254126 TI - Multiple Toll-like receptor ligands induce an IL-6 transcriptional response in skeletal myocytes. AB - Toll-like receptors (TLRs) comprise a critical sentinel that monitors body compartments for the presence of pathogens. Skeletal muscle expresses TLRs and responds to pathogen-associated molecular patterns (PAMPs), such as lipopolysaccharide (LPS), by mounting an innate immune response. In the present study, we used C2C12 myocytes as a model system for skeletal muscle during infection. C2C12 cells responded to LPS in a time frame and with a pattern of gene expression that faithfully mimicked the response of skeletal muscle to LPS in vivo. LPS from a variety of Escherichia coli serotypes stimulated IL-6 synthesis. C2C12 cells expressed TLR1-7, but not TLR8 or TLR9, mRNA by RT-PCR. A synthetic tripalmitoylated cysteine-, serine-, and lysine-containing peptide (Pam) and LPS from Porphyromonas gingivalis, two TLR2 ligands, also stimulated IL 6 expression. LPS and Pam stimulated luciferase activity driven from NF-kappaB and IL-6 promoter-containing plasmids, and this response was blunted when the NF kappaB binding site was mutated. LPS- and Pam-stimulated IL-6 expression was inhibited by the proteasome inhibitor MG-132 and the IkappaB kinase-2 (IKK2) inhibitor 2-[(aminocarbonyl)amino]-5-(4-fluorophenyl)-3-thiophenecarboxamide (TPCA-1). Pam-stimulated NF-kappaB and IL-6 promoter activities were disrupted by a dominant-negative form of TLR2, but not TLR4. Local injection of LPS or Pam into the gastrocnemius muscle stimulated IL-6 mRNA expression in the injected, but not the contralateral, muscle. The LPS- but not Pam-stimulated expression of IL-6 mRNA was blunted in skeletal muscle of mice carrying an inactivating mutation in TLR4. The data suggest that skeletal muscle and muscle cells recognize pathogen-associated molecules with specific TLRs to initiate an IL-6 transcriptional response. PMID- 16254127 TI - Adenoviral PR39 improves blood flow and myocardial function in a pig model of chronic myocardial ischemia by enhancing collateral formation. AB - Angiogenic therapy with individual growth factors or "master switch" genes is being evaluated for treatment of advanced coronary artery disease. In this study, we investigated the efficacy and mechanism of PR39, a gene capable of activating VEGF and fibroblast growth factor (FGF)-2-dependent pathways. PR39 enhances hypoxia-inducible factor-1alpha (HIF-1alpha)-dependent gene expression by selectively inhibiting proteasome degradation of this transcription factor. In addition, PR39 also stimulates expression of the FGF receptors (FGFR)-1 and syndecan-4. In a pig model of chronic myocardial ischemia, we used angiography, MRI, and microsphere regional blood flow to evaluate the efficacy of intramyocardial adenoviral protein arginine-rich peptide (Ad-PR39) injections. Ad PR39 improved collateral scores, regional perfusion, and regional function in a dose-dependent manner. Local VEGF, VEGFR-1, VEGFR-2, syndecan, and FGFR-1 levels were 16-75% upregulated after Ad-PR39 injections as assessed by real-time PCR, suggesting upregulation of VEGF and FGF pathways. PR39 is an angiogenic peptide that improves perfusion and function of ischemic myocardium, at least in part, through collateral formation. The dual mechanism, i.e., stimulation of HIF-1alpha and FGF receptor expression, likely accounts for the functional benefits of PR39. PMID- 16254128 TI - Chronic excessive erythrocytosis induces endothelial activation and damage in mouse brain. AB - Excessive erythrocytosis results in severely increased blood viscosity, which may have significant detrimental effects on endothelial cells and, ultimately, function of the vascular endothelium. Because blood-brain barrier stability is crucial for normal physiological function, we used our previously characterized erythropoietin-overexpressing transgenic (tg6) mouse line (which has a hematocrit of 0.8-0.9) to investigate the effect of excessive erythrocytosis on vessel number, structure, and integrity in vivo. These mice have abnormally high levels of nitric oxide (NO), a potent proinflammatory molecule, suggesting altered vascular permeability and function. In this study, we observed that brain vessel density of tg6 mice was significantly reduced (16%) and vessel diameter was significantly increased (15%) compared with wild-type mice. Although no significant increases in vascular permeability under normoxic or acute hypoxic conditions (8% O2 for 4 h) were detected, electron-microscopic analysis revealed altered morphological characteristics of the tg6 endothelium. Tg6 brain vascular endothelial cells appeared to be activated, with increased luminal protrusions reminiscent of ongoing inflammatory processes. Consistent with this observation, we detected increased levels of intercellular adhesion molecule-1 and von Willebrand factor, markers of endothelial activation and damage, in brain tissue. We propose that chronic excessive erythrocytosis and sustained high hematocrit cause endothelial damage, which may, ultimately, increase susceptibility to vascular disease. PMID- 16254129 TI - Restraint stress augments postprandial gastric contractions but impairs antropyloric coordination in conscious rats. AB - Central corticotropin-releasing factor (CRF) plays an important role in mediating restraint stress-induced delayed gastric emptying. However, it is unclear how restraint stress modulates gastric motility to delay gastric emptying. Inasmuch as solid gastric emptying is regulated via antropyloric coordination, we hypothesized that restraint stress impairs antropyloric coordination, resulting in delayed solid gastric emptying in conscious rats. Two strain gauge transducers were sutured onto the serosal surface of the antrum and pylorus, and postprandial gastric motility was monitored before, during, and after restraint stress. Antropyloric coordination, defined as a propagated single contraction from the antrum to the pylorus within 10 s, was followed by > or = 20 s of quiescence. Restraint stress enhanced postprandial gastric motility in the antrum and pylorus to 140 +/- 9% and 134 +/- 9% of basal, respectively (n = 6). The number of episodes of antropyloric coordination before restraint stress, 2.4 +/- 0.4/10 min, was significantly reduced to 0.6 +/- 0.3/10 min by restraint stress. Intracisternal injection of the CRF type 2 receptor antagonist astressin 2B (60 microg) or guanethidine partially restored restraint stress-induced impairment of antropyloric coordination (1.6 +/- 0.3/10 min, n = 6). The restraint stress induced augmentation of antral and pyloric contractions was increased by astressin 2B and guanethidine but abolished by atropine, hexamethonium, and vagotomy. Restraint stress enhanced postprandial gastric motility via a vagal cholinergic pathway. Restraint stress-induced delay of solid gastric emptying is due to impairment of antropyloric coordination. Restraint stress-induced impairment of antropyloric coordination might be mediated via a central CRF pathway. PMID- 16254130 TI - Patients with solid tumors treated with high-temperature whole body hyperthermia show a redistribution of naive/memory T-cell subtypes. AB - An activation of the immune system might contribute to the therapeutic effect of whole body hyperthermia (WBH) in cancer patients. We explored immune and endocrine responses in patients undergoing high-temperature WBH. Identical parameters were investigated in a separate group of healthy volunteers undergoing physical exercise to rule out effects of sympathetic activation. Lymphocyte subpopulations, lymphocytic expression of a range of adhesion molecules, and serum concentrations of a variety of hormones and cytokines were assessed in cancer patients undergoing high-temperature (60 min at 41.0-41.8 degrees C) WBH (n = 25) and in a separate group of healthy volunteers (n = 10) performing strenuous physical exercise. WBH induced an increase in human growth hormone (hGH), ACTH, and cortisol as well as in TNF-alpha, IL-6, IL-8, and IL-12R. We observed an increase in natural killer (NK) cells and CD56+ NK T cells shortly after initiation of WBH. In contrast, we found a decrease in T cells expressing L selectin (CD62L) or alpha4beta7 integrin adhesion molecules mediating homing to lymphatic tissues. Accordingly, we observed a decrease in CD45RA+CCR7+ naive and CD45RA-CCR7+ central memory T cells. Numbers of CD45RA-CCR7- memory effector and CD45RA+CCR7 terminally differentiated T cells, on the other hand, remained unchanged. No comparable changes were observed in the group of healthy volunteers. In conclusion, patients with solid tumors treated with WBH show an increase in NK and NK T cells. In a later phase, plasma concentrations of IL-8, hGH, and cortisol increase, correlated with an influx of neutrophils into the peripheral blood. The alterations in T-cell populations suggest that WBH may induce naive and central-memory T cells to enter lymphatic tissue to await antigen exposure and effector T cells to migrate into peripheral tissues to exert their effector function. Although the exercise group may not be an appropriate control to proof the effect of WBH, these changes were not seen in the healthy volunteers performing physical exercise. PMID- 16254131 TI - High-efficacy 5-hydroxytryptamine 1A receptor activation counteracts opioid hyperallodynia and affective conditioning. AB - Pain may become intractable as tolerance develops to opioids and the opioids, paradoxically, induce pain. We examined the hypothesis that the analgesia produced by the novel analgesic and high-efficacy 5-hydroxytryptamine (5-HT)(1A) receptor agonist (3-chloro-4-fluoro-phenyl)-[4-fluoro-4-[[(5-methyl-pyridin-2 ylmethyl)-amino]methyl]piperidin-1-yl]methanone, fumaric acid salt (F 13640) may counteract opioid-induced pain. In studies of the somatosensory quality of pain in infraorbital nerve-injured rats, morphine infusion (5 mg/day) by means of osmotic pumps initially caused analgesia (i.e., decreased the behavioral response to von Frey filament stimulation), followed by hyperallodynia and analgesic tolerance. Infusion of F 13640 (0.63 mg/day) prevented the development of opioid hyperallodynia and reversed opioid hyperallodynia once established. In studies of the affective/motivational quality of pain, F 13640 both prevented and reversed the conditioned place aversion induced by naloxone (0.04 mg/kg i.p.) in morphine infused rats; F 13640 also prevented and reversed the conditioned place preference induced by morphine injections (7.5 mg/kg i.p.). The data confirm that opioids produce bidirectional hypo- and proalgesic actions, and offer initial evidence that high-efficacy 5-HT(1A) receptor activation counteracts both the sensory and the affective/motivational qualities of opioid-induced pain. The data also indicate that F 13640 may be effective with opioid-resistant pain. It further is suggested that opioid addiction may represent self-therapy of opioid induced pathological pain. PMID- 16254132 TI - The relaxant activity of 4,7-dimethyl-1,2,5-oxadiazolo[3,4-d]-pyridazine 1,5,6 trioxide in the mouse corpus cavernosum. AB - We have studied the effect of an activator of soluble guanylate cyclase 4,7 dimethyl-1,2,5-oxadiazolo[3,4-d]pyridazine 1,5,6-trioxide (FPTO) on the tone and nitrergic relaxation responses of mouse cavernous strips and compared FPTO to a known nitric oxide donor sodium nitroprusside. FPTO thiol-dependently generated nitric oxide measured by polarography and activated purified human soluble guanylate cyclase. FPTO and sodium nitroprusside relaxed the cavernous tissue in a concentration-dependent manner. A nitric-oxide synthase inhibitor N(omega) nitro-L-arginine did not alter the relaxations to FPTO or sodium nitroprusside, whereas soluble guanylate cyclase inhibitor 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin 1-one (ODQ) suppressed relaxation to FPTO and sodium nitroprusside. Exogenously added thiols L-cysteine or dithiothreitol inhibited the relaxant responses to FPTO but not to sodium nitroprusside, whereas glutathione did not influence the responses to both agents. Thiol alkylation agent N-ethylmaleimide significantly enhanced FPTO-induced relaxation, and thiol-modifying agent diamide inhibited relaxation to FPTO. The potentiating effect of N-ethylmaleimide was neutralized by coadministration of N-ethylmaleimide with glutathione, L-cysteine, dithiothreitol, or ODQ. N-Ethylmaleimide but not diamide significantly inhibited relaxation induced by sodium nitroprusside. FPTO potently suppressed contraction to electrical field stimulation, which was prevented by glutathione or L cysteine. In addition, FPTO did not affect relaxation produced by electrical field stimulation in phenylephrine-precontracted tissue. Our results show that FPTO can relax mouse corpus cavernosum and that the relaxant activity of this agent is thiol- and soluble guanylate cyclase-dependent. This effect could be potentiated by N-ethylmaleimide. FPTO does not potentiate nitrergic relaxation induced by electrical field stimulation. PMID- 16254133 TI - Selective inhibition of phosphodiesterase-4 ameliorates chronic colitis and prevents intestinal fibrosis. AB - The phosphodiesterase-4 (PDE4) inhibitors may be an important target in the treatment of several inflammatory conditions. The anti-inflammatory effect of PDE4 inhibitors bears similarities with that of steroids, without interfering with the hypophysary-adrenal-axis. We compared the effect of rolipram, a selective PDE4 inhibitor, with steroids on the clinical course of experimental colitis induced by 2,4,6-trinitrobenzenesulfonic acid (TNBS). Three groups of rats (n = 20) received TNBS. One group received methylprednisolone from day 7, another group received rolipram from the same day, and control group received no further treatment. On days 14 and 21 after TNBS instillation, sets of 10 rats underwent colonic dialysis to measure eicosanoid release. Colonic lesions were blindly scored, and colons were homogenized for quantification of myeloperoxidase (MPO) activity and collagen content. Concentration of tumor necrosis factor alpha (TNF-alpha) and transforming growth factor beta1 (TGF-beta1) in colonic tissue was also measured. Both treatments reduced significantly the eicosanoid release and MPO activity. On day 14, both rolipram and methylprednisolone significantly reduced TNF-alpha content, but TGF-beta1 was only inhibited by rolipram. On day 21, lesion scores and collagen content were significantly reduced only in rolipram-treated group. In conclusion, PDE4 inhibition by rolipram markedly ameliorates the course of chronic colitis and it is superior to methylprednisolone in preventing late collagen deposition. PMID- 16254134 TI - KIT-D816 mutations in AML1-ETO-positive AML are associated with impaired event free and overall survival. AB - Mutations in codon D816 of the KIT gene represent a recurrent genetic alteration in acute myeloid leukemia (AML). To clarify the biologic implication of activation loop mutations of the KIT gene, 1940 randomly selected AML patients were analyzed. In total, 33 (1.7%) of 1940 patients were positive for D816 mutations. Of these 33 patients, 8 (24.2%) had a t(8;21), which was significantly higher compared with the subgroup without D816 mutations. Analyses of genetic subgroups showed that KIT-D816 mutations were associated with t(8;21)/AML1-ETO and other rare AML1 translocations. In contrast, other activating mutations like FLT3 and NRAS mutations were very rarely detected in AML1-rearranged leukemia. KIT mutations had an independent negative impact on overall (median 304 vs 1836 days; P = .006) and event-free survival (median 244 vs 744 days; P = .003) in patients with t(8;21) but not in patients with a normal karyotype. The KIT-D816V receptor expressed in Ba/F3 cells was resistant to growth inhibition by the selective PTK inhibitors imatinib and SU5614 but fully sensitive to PKC412. Our findings clearly indicate that activating mutations of receptor tyrosine kinases are associated with distinct genetic subtypes in AML. The KIT-D816 mutations confer a poor prognosis to AML1-ETO-positive AML and should therefore be included in the diagnostic workup. Patients with KIT-D816-positive/AML1-ETO-positive AML might benefit from early intensification of treatment or combination of conventional chemotherapy with KIT PTK inhibitors. PMID- 16254135 TI - Radiofrequency ablation of lymphoma. AB - Percutaneous minimally invasive radiofrequency (RF) ablation has not been described for lymphoma. This image-guided modality is presented in 3 different settings for the treatment of refractory lymphoma. The first patient received RF ablation for the curative treatment of a solitary residual hepatic mass following rituximab-based chemotherapy for a posttransplantation lymphoproliferative disorder (PTLD) and is disease-free 4 years later. The second patient received RF ablation for successful palliation of progressive follicular lymphoma adjacent to the bladder wall following chemotherapy and maximum radiation. The third patient received RF ablation for prevention of airway obstruction from progressive diffuse large B-cell lymphoma of the right neck following chemotherapy and maximum radiation. RF ablation may be clinically beneficial and should be considered for the treatment of local lymphoma that is refractory or not amenable to standard approaches. PMID- 16254137 TI - Anaplastic lymphoma kinase activity is essential for the proliferation and survival of anaplastic large-cell lymphoma cells. AB - The roles of aberrant expression of constitutively active ALK chimeric proteins in the pathogenesis of anaplastic large-cell lymphoma (ALCL) have been well defined; nevertheless, the notion that ALK is a molecular target for the therapeutic modulation of ALK+ ALCL has not been validated thus far. Select fused pyrrolocarbazole (FP)-derived small molecules with ALK inhibitory activity were used as pharmacologic tools to evaluate whether functional ALK is essential for the proliferation and survival of ALK+ ALCL cells in culture. These compounds inhibited interleukin 3 (IL-3)-independent proliferation of BaF3/NPM-ALK cells in an ALK inhibition-dependent manner and significantly blocked colony formation in agar of mouse embryonic fibroblast (MEF) cells harboring NPM-ALK. Inhibition of NPM-ALK phosphorylation in the ALK+ ALCL-derived cell lines resulted in significant inhibition of cell proliferation and induction of apoptotic-cell death, while having marginal effects on the proliferation and survival of K562, an ALK- leukemia cell line. ALK inhibition resulted in cell-cycle G1 arrest and inactivation of ERK1/2, STAT3, and AKT signaling pathways. Potent and selective ALK inhibitors may have therapeutic application for ALK+ ALCL and possibly other solid and hematologic tumors in which ALK activation is implicated in their pathogenesis. PMID- 16254138 TI - The inhibitory receptor IRp60 (CD300a) suppresses the effects of IL-5, GM-CSF, and eotaxin on human peripheral blood eosinophils. AB - Allergic, inflammatory, and immune responses carried out by eosinophils are regulated by the cross talk between activatory and inhibitory signals. While much data has been obtained on activatory signals, inhibitory receptors on these cells have received scant attention. Therefore, we screened the surface of human peripheral blood eosinophils for inhibitory receptors using monoclonal antibodies (mAbs) previously generated to recognize receptors on human natural killer cells. Eosinophils from all of the donors examined expressed the inhibitory receptors IRp60, LIR3/ILT5, FcgammaRIIB, and p75/AIRM but not LIR1/ILT2, p58.1, p58.2, p70, or NKG2A/CD94 (n = 15). Interestingly, 25% of the donors expressed p140. IRp60 cross-linking inhibited eotaxin-dependent transmigration of eosinophils in a calcium-independent fashion. In addition, cross-linking of IRp60 on the eosinophils in the presence of IL-5/GM-CSF inhibited the antiapoptotic effect of these cytokines and blocked the release of TNF-alpha, IL-1beta, IFN-gamma, IL-4, and 3T3 fibroblast proliferation. Cross-linking of IRp60 inhibited IL-5-mediated JAK2 phosphorylation as well as eotaxin- and IL-5/GM-CSF-mediated ERK1/2 and p38 phosphorylation. Furthermore, upon cross-linking, IRp60 underwent tyrosine phosphorylation and recruited SHP-1 but not SHP-2. These findings demonstrate a novel pathway for suppressing the activity of human eosinophils, thus indicating IRp60 as a future potential target for the treatment of allergic and eosinophil associated diseases. PMID- 16254136 TI - Designer blood: creating hematopoietic lineages from embryonic stem cells. AB - Embryonic stem (ES) cells exhibit the remarkable capacity to become virtually any differentiated tissue upon appropriate manipulation in culture, a property that has been beneficial for studies of hematopoiesis. Until recently, the majority of this work used murine ES cells for basic research to elucidate fundamental properties of blood-cell development and establish methods to derive specific mature lineages. Now, the advent of human ES cells sets the stage for more applied pursuits to generate transplantable cells for treating blood disorders. Current efforts are directed toward adapting in vitro hematopoietic differentiation methods developed for murine ES cells to human lines, identifying the key interspecies differences in biologic properties of ES cells, and generating ES cell-derived hematopoietic stem cells that are competent to repopulate adult hosts. The ultimate medical goal is to create patient-specific and generic ES cell lines that can be expanded in vitro, genetically altered, and differentiated into cell types that can be used to treat hematopoietic diseases. PMID- 16254139 TI - Redirecting differentiation of hematopoietic progenitors by a transcription factor, GATA-2. AB - GATA-2 is a zinc finger transcription factor essential for differentiation of immature hematopoietic cells. We analyzed the function of GATA-2 by a combined method of tetracycline-dependent conditional gene expression and in vitro hematopoietic differentiation from mouse embryonic stem (ES) cells using OP9 stroma cells (OP9 system). In the presence of macrophage colony-stimulating factor (M-CSF), the OP9 system induced macrophage differentiation. GATA-2 expression in this system inhibited macrophage differentiation and redirected the fate of hematopoietic differentiation to other hematopoietic lineages. GATA-2 expression commencing at day 5 or day 6 induced megakaryocytic or erythroid differentiation, respectively. Expression levels of PU.1, a hematopoietic transcription factor that interferes with GATA-2, appeared to play a critical role in differentiation to megakaryocytic or erythroid lineages. Transcription of PU.1 was affected by histone acetylation induced by binding of GATA-2 to the PU.1 promoter region. This study demonstrates that the function of GATA-2 is modified in a context-dependent manner by expression of PU.1, which in turn is regulated by GATA-2. PMID- 16254140 TI - 131I-anti-CD45 antibody plus busulfan and cyclophosphamide before allogeneic hematopoietic cell transplantation for treatment of acute myeloid leukemia in first remission. AB - In an attempt to improve outcomes for patients with acute myeloid leukemia (AML) after allogeneic hematopoietic cell transplantation (HCT), we conducted a phase 1/2 study in which targeted irradiation delivered by 131I-anti-CD45 antibody was combined with targeted busulfan (BU; area-under-curve, 600-900 ng/mL) and cyclophosphamide (CY; 120 mg/kg). Fifty-two (88%) of 59 patients receiving a trace 131I-labeled dose of 0.5 mg/kg anti-CD45 murine antibody had higher estimated absorbed radiation in bone marrow and spleen than in any other organ. Forty-six patients were treated with 102 to 298 mCi (3774-11 026 MBq) 131I, delivering an estimated 5.3 to 19 (mean, 11.3) Gy to marrow, 17-72 (mean, 29.7) Gy to spleen, and 3.5 Gy (n = 4) to 5.25 Gy (n = 42) to the liver. The estimated 3-year nonrelapse mortality and disease-free survival (DFS) were 21% and 61%, respectively. These results were compared with those from 509 similar International Bone Marrow Transplant Registry patients who underwent transplantation using BU/CY alone. After adjusting for differences in age and cytogenetics risk, the hazard of mortality among all antibody-treated patients was 0.65 times that of the Registry patients (95% CI 0.39-1.08; P = .09). The addition of targeted hematopoietic irradiation to conventional BU/CY is feasible and well tolerated, and phase 2 results are sufficiently encouraging to warrant further study. PMID- 16254141 TI - AKT induces erythroid-cell maturation of JAK2-deficient fetal liver progenitor cells and is required for Epo regulation of erythroid-cell differentiation. AB - AKT serine threonine kinase of the protein kinase B (PKB) family plays essential roles in cell survival, growth, metabolism, and differentiation. In the erythroid system, AKT is known to be rapidly phosphorylated and activated in response to erythropoietin (Epo) engagement of Epo receptor (EpoR) and to sustain survival signals in cultured erythroid cells. Here we demonstrate that activated AKT complements EpoR signaling and supports erythroid-cell differentiation in wild type and JAK2-deficient fetal liver cells. We show that erythroid maturation of AKT-transduced cells is not solely dependent on AKT-induced cell survival or proliferation signals, suggesting that AKT transduces also a differentiation specific signal downstream of EpoR in erythroid cells. Down-regulation of expression of AKT kinase by RNA interference, or AKT activity by expression of dominant negative forms, inhibits significantly fetal liver-derived erythroid cell colony formation and gene expression, demonstrating that AKT is required for Epo regulation of erythroid-cell maturation. PMID- 16254142 TI - Zebrafish to humans: evolution of the alpha3-chain of type IV collagen and emergence of the autoimmune epitopes associated with Goodpasture syndrome. AB - Goodpasture syndrome is an autoimmune vascular disease associated with kidney and lung failure, with pathogenic circulating autoantibodies targeted to a set of discontinuous epitope sequences within the noncollagenous domain-1 (NC1) of the alpha3 chain of type IV collagen (alpha3(IV)NC1), the Goodpasture autoantigen. We demonstrate that basement membrane extracted NC1 domain preparations from Caenorhabditis elegans, Drosophila melanogaster, and Danio rerio do not bind Goodpasture autoantibodies, while Xenopus laevis, chicken, mouse and human alpha3(IV)NC1 domains bind autoantibodies. The alpha3(IV) chain is not present in C elegans and Drosophila melanogaster, but is first detected in the Danio rerio. Interestingly, native Danio rerio alpha3(IV)NC1 does not bind Goodpasture autoantibodies. Next, we cloned, sequenced, and generated recombinant Danio rerio alpha3(IV)NC1 domain. In contrast to recombinant human alpha3(IV)NC1 domain, there was complete absence of autoantibody binding to recombinant Danio rerio alpha3(IV)NC1. Three-dimensional molecular modeling from existing x-ray coordinates of human NC1 domain suggest that evolutionary alteration of electrostatic charge and polarity due to the emergence of critical serine, aspartic acid, and lysine residues, accompanied by the loss of asparagine and glutamine, contributes to the emergence of the 2 major Goodpasture epitopes on the human alpha3(IV)NC1 domain, as it evolved from the Danio rerio over 450 million years. PMID- 16254143 TI - Efficacy and safety of rituximab in B-cell post-transplantation lymphoproliferative disorders: results of a prospective multicenter phase 2 study. AB - B-cell posttransplantation lymphoproliferative disorder (B-PTLD) is a rare but severe complication of transplantation, with no consensus on best treatment practice. This prospective trial, the first to test a treatment for PTLD, was designed to evaluate the efficacy and safety of rituximab in patients with B-PTLD after solid organ transplantation (SOT). Forty-six patients were included and 43 patients were analyzed. Patients were eligible if they had untreated B-PTLD that was not responding to tapering of immunosuppression. Treatment consisted of 4 weekly injections of rituximab at 375 mg/m2. At day (d) 80, 37 (86%) patients were alive, and the response rate was 44.2%, including 12 complete response/unconfirmed complete response (CR/CRu). The only factor predictive of a response at d80 was a normal lactate dehydrogenase level (P = .007, odds ratio [OR] = 6.9). At d360, responses were maintained in 68% of patients, and 56% of patients were alive. The overall survival rate at 1 year was 67%. We conclude that rituximab is effective and safe in PTLD, with stable responses at 1 year. The response rate and overall survival might be improved by combining rituximab with other treatments. PMID- 16254144 TI - The epidemiology of aplastic anemia in Thailand. AB - Aplastic anemia has been linked to environmental exposures, from chemicals and medical drugs to infectious agents. The disease occurs more frequently in Asia than in the West, with incidence rates 2- to 3-fold higher. We report updated results of an epidemiologic study conducted in Thailand from 1989 to 2002, in which 541 patients and 2261 controls were enrolled. Exposures were determined by in-person interview. We observed significantly elevated relative risk estimates for benzene (3.5) and other solvents (2.0) and for sulfonamides (5.6), thiazides (3.8), and mebendazole (3.0). Chloramphenicol use was infrequent, and no significant association was observed. Agricultural pesticides were implicated in Khonkaen (northeastern Thailand). There were significant associations with organophosphates (2.1), DDT (6.7), and carbamates (7.4). We found significant risks for farmers exposed to ducks and geese (3.7) and a borderline association with animal fertilizer (2.1). There was a significant association in Khonkaen with drinking other than bottled or distilled water (2.8). Nonmedical needle exposure was associated in Bangkok and Khonkaen combined (3.8). Most striking was the large etiologic fraction in a rural region accounted for by animal exposures and drinking of water from sources such as wells, rural taps, and rainwater, consistent with an infectious etiology for many cases of aplastic anemia in Thailand. PMID- 16254145 TI - Regulation of survivin expression through Bcr-Abl/MAPK cascade: targeting survivin overcomes imatinib resistance and increases imatinib sensitivity in imatinib-responsive CML cells. AB - KBM5 cells, derived from a patient with blast crisis Philadelphia chromosome positive (Ph+) chronic myelogenous leukemia (CML), and imatinib-resistant KBM5 (KBM5-STI571) cells were found to express high levels of survivin. Inhibition of Bcr-Abl by imatinib significantly decreased survivin expression and cell viability in KBM5, but much less so in KBM5-STI571 cells. Inhibition of MEK, downstream of the Bcr-Abl signaling cascade decreased survivin expression and cell viability in both KBM5 and KBM5-STI571 cells. In addition, down-regulation of survivin by a survivin antisense oligonucleotide (Sur-AS-ODN) inhibited cell growth and induced maximal G2M block at 48 hours, whereas cell death was observed only at 72 hours in both KBM5 and KBM5-STI571 cells as shown by annexin V staining. Further, the combination of Sur-AS-ODN and imatinib induced more cell death in KBM5 cells than did either treatment alone. Down-regulating survivin also decreased colony-forming units (CFUs) in blast crisis CML patient samples. Our data therefore suggest that survivin is regulated by the Bcr-Abl/MAPK cascade in Ph+ CML. The facts that down-regulating survivin expression induced cell growth arrest and subsequent cell death regardless of the cell response to imatinib and enhanced the sensitivity to imatinib suggest the potential therapeutic utility of this strategy in patients with CML, both imatinib sensitive and resistant. PMID- 16254146 TI - MHC class II expression through a hitherto unknown pathway supports T helper cell dependent immune responses: implications for MHC class II deficiency. AB - MHC class II (MHCII) deficiency or bare lymphocyte syndrome (BLS) is a severe immunodeficiency characterized by deficient T helper (Th)-cell-dependent immunity. The disease is caused by defects of the MHCII promoter complex resulting in low or absent MHCII expression. We demonstrate in a murine model of MHCII deficiency (RFX5- or CIITA-deficient mice) that residual MHCII expression by professional antigen-presenting cells (APCs) is sufficient to support activation of adoptively transferred Th cells. Furthermore, upon transplantation of WT thymic epithelium, we observed development of endogenous Th cells with restoration of Th-cell-dependent antibody responses and immunity to cytomegalovirus infection, thus opening the possibility of an alternative treatment regimen for BLS. Residual MHCII expression was further induced by the presence of Th cells and also other stimuli. Analysis of CIITA/RFX5 double deficient animals revealed that this inducible MHCII expression is genetically independent of the known promoter complex and thus constitutes an alternative MHCII expression pathway. In these experiments, we also detected a novel repressive function of the RFX complex in the absence of CIITA. PMID- 16254148 TI - Logic of the yeast metabolic cycle: temporal compartmentalization of cellular processes. AB - Budding yeast grown under continuous, nutrient-limited conditions exhibit robust, highly periodic cycles in the form of respiratory bursts. Microarray studies reveal that over half of the yeast genome is expressed periodically during these metabolic cycles. Genes encoding proteins having a common function exhibit similar temporal expression patterns, and genes specifying functions associated with energy and metabolism tend to be expressed with exceptionally robust periodicity. Essential cellular and metabolic events occur in synchrony with the metabolic cycle, demonstrating that key processes in a simple eukaryotic cell are compartmentalized in time. PMID- 16254147 TI - Randomized use of cyclosporin A (CsA) to modulate P-glycoprotein in children with AML in remission: Pediatric Oncology Group Study 9421. AB - Relapse is a major obstacle in the cure of acute myeloid leukemia (AML). The Pediatric Oncology Group AML Study 9421 tested 2 different strategies to improve event-free survival (EFS) and overall survival (OS). Patients were randomized to receive standard-dose DAT (daunorubicin, cytarabine, and thioguanine) or high dose DAT during induction. To interfere with P-glycoprotein (P-gp)-dependent drug efflux, the second randomization tested the benefit of cyclosporine (CsA) added to consolidation chemotherapy. Of the 282 children randomly assigned to receive standard DAT induction, 248 (87.9%) achieved remission compared to 253 (91%) of the 278 receiving high-dose DAT (P = ns). Children with HLA-identical sibling donors who achieved a complete remission received an allogeneic bone marrow transplant as consolidation. For the 83 patients receiving a matched related donor bone marrow transplantation (BMT), the 3-year disease-free survival (DFS) is 67%. Of the 418 children who achieved remission and went on to consolidation with and without CsA, the DFS was 40.6% and 33.9%, respectively (P = .24). Overexpression of P-gp was infrequent (14%) in this pediatric population. In this study, intensifying induction with high-dose DAT and the addition of CsA to consolidation chemotherapy did not prolong the durations of remission or improve overall survival for children with AML. PMID- 16254149 TI - Golgi duplication in Trypanosoma brucei requires Centrin2. AB - Centrins are highly conserved components of the centrosome, which in the parasitic protozoan T. brucei comprises the basal body and nucleates the flagellum used for locomotion. Here, we found TbCentrin2 in an additional bi lobed structure near to the Golgi apparatus. One lobe was associated with the old Golgi, and the other became associated with the newly forming Golgi as the cell grew. Depletion of TbCentrin1 inhibited duplication of the basal body, whereas depletion of TbCentrin2 also inhibited duplication of the Golgi. Thus, a Centrin2 containing structure distinct from the basal body appears to mark the site for new Golgi assembly. PMID- 16254150 TI - A thiolate-ligated nonheme oxoiron(IV) complex relevant to cytochrome P450. AB - Thiolate-ligated oxoiron(IV) centers are postulated to be the key oxidants in the catalytic cycles of oxygen-activating cytochrome P450 and related enzymes. Despite considerable synthetic efforts, chemists have not succeeded in preparing an appropriate model complex. Here we report the synthesis and spectroscopic characterization of [FeIV(O)(TMCS)]+ where TMCS is a pentadentate ligand that provides a square pyramidal N4(SR)apical, where SR is thiolate, ligand environment about the iron center, which is similar to that of cytochrome P450. The rigidity of the ligand framework stabilizes the thiolate in an oxidizing environment. Reactivity studies suggest that thiolate coordination favors hydrogen-atom abstraction chemistry over oxygen-atom transfer pathways in the presence of reducing substrates. PMID- 16254151 TI - Ecosystem service supply and vulnerability to global change in Europe. AB - Global change will alter the supply of ecosystem services that are vital for human well-being. To investigate ecosystem service supply during the 21st century, we used a range of ecosystem models and scenarios of climate and land use change to conduct a Europe-wide assessment. Large changes in climate and land use typically resulted in large changes in ecosystem service supply. Some of these trends may be positive (for example, increases in forest area and productivity) or offer opportunities (for example, "surplus land" for agricultural extensification and bioenergy production). However, many changes increase vulnerability as a result of a decreasing supply of ecosystem services (for example, declining soil fertility, declining water availability, increasing risk of forest fires), especially in the Mediterranean and mountain regions. PMID- 16254152 TI - Chess game at the FDA. PMID- 16254154 TI - Astronomy. Science takes back seat as Hubble shoots the Moon. PMID- 16254155 TI - Genomics. New haplotype map may overhaul gene hunting. PMID- 16254153 TI - Scientific community. 'Security breach' leaks NIH grant applications onto Web. PMID- 16254156 TI - Neuroscience. Does brain cell growth drive weight loss? PMID- 16254157 TI - Science policy. French agency cited for lack of women. PMID- 16254158 TI - Medicine. Fused genes may help explain the origins of prostate cancer. PMID- 16254159 TI - European science. Aggrieved Turkish scientists welcome an E.U. review. PMID- 16254160 TI - U.S. workforce. A glass ceiling for Asian scientists? PMID- 16254161 TI - Profile. South Africa's bone man: 80 and still digging into the past. PMID- 16254162 TI - Antarctic drilling. The plan to unlock Lake Vostok. PMID- 16254163 TI - Archaeology. 'Deviant' burials reveal death on the fringe in ancient societies. PMID- 16254164 TI - Astronomy. Astronomers sweep space for the sources of cosmic dust. PMID- 16254166 TI - Revisiting the Grand Canyon. PMID- 16254165 TI - A plea to save the Voyager mission. PMID- 16254167 TI - Deja vu all over again for nuclear power? PMID- 16254168 TI - Issues surrounding nuclear power. PMID- 16254169 TI - The benefits of solar thermal energy. PMID- 16254170 TI - A "chick-a-dee" or a "co-qui"? PMID- 16254171 TI - Regulating commercial cloning of animals. PMID- 16254172 TI - Density is only relative. PMID- 16254173 TI - Policy. OPV cessation--the final step to a "polio-free" world. AB - The 20-year, U.S. $4 billion-dollar international effort to eradicate wild polioviruses now includes monovalent oral poliovirus type 1 vaccine (mOPV1), which was first given to 40 million children in India in April 2005. As this vaccine is being introduced to eliminate some of the last poliovirus reservoirs, the Global Polio Eradication Initiative is planning for the eventual synchronized worldwide cessation of the routine use of all oral poliovirus vaccines (OPVs) because of their capacity to cause, rarely, outbreaks of paralytic poliomyelitis. This Policy Forum reviews the feasibility and implications of eventually stopping the use of live polio vaccines. PMID- 16254174 TI - AtmospheRIC science. Tipping points in the tundra. PMID- 16254175 TI - Virology. What links bats to emerging infectious diseases? PMID- 16254176 TI - Developmental biology. Less steroids make bigger flies. PMID- 16254177 TI - Physics. The observation of matter wave fluctuations. PMID- 16254178 TI - Retrospective. Joseph Rotblat (1908-2005). PMID- 16254179 TI - Progress in modeling of protein structures and interactions. AB - The prediction of the structures and interactions of biological macromolecules at the atomic level and the design of new structures and interactions are critical tests of our understanding of the interatomic interactions that underlie molecular biology. Equally important, the capability to accurately predict and design macromolecular structures and interactions would streamline the interpretation of genome sequence information and allow the creation of macromolecules with new and useful functions. This review summarizes recent progress in modeling that suggests that we are entering an era in which high resolution prediction and design will make increasingly important contributions to biology and medicine. PMID- 16254180 TI - The Asian tsunami: a protective role for coastal vegetation. AB - The 26 December 2004 Indian Ocean tsunami had major effects on coastal communities and ecosystems. An assessment of coastlines after the tsunami indicates that coastal vegetation such as mangroves and beach forests helped to provide protection and reduce effects on adjacent communities. In recent years, mangroves and other coastal vegetation have been cleared or degraded along many coastlines, increasing their vulnerability to storm and tsunami damage. Establishing or strengthening greenbelts of mangroves and other coastal forests may play a key role in reducing the effect of future extreme events. PMID- 16254182 TI - Quantum coherence in an optical modulator. AB - Semiconductor quantum well electroabsorption modulators are widely used to modulate near-infrared (NIR) radiation at frequencies below 0.1 terahertz (THz). Here, the NIR absorption of undoped quantum wells was modulated by strong electric fields with frequencies between 1.5 and 3.9 THz. The THz field coupled two excited states (excitons) of the quantum wells, as manifested by a new THz frequency- and power-dependent NIR absorption line. Nonperturbative theory and experiment indicate that the THz field generated a coherent quantum superposition of an absorbing and a nonabsorbing exciton. This quantum coherence may yield new applications for quantum well modulators in optical communications. PMID- 16254181 TI - Recurrent fusion of TMPRSS2 and ETS transcription factor genes in prostate cancer. AB - Recurrent chromosomal rearrangements have not been well characterized in common carcinomas. We used a bioinformatics approach to discover candidate oncogenic chromosomal aberrations on the basis of outlier gene expression. Two ETS transcription factors, ERG and ETV1, were identified as outliers in prostate cancer. We identified recurrent gene fusions of the 5' untranslated region of TMPRSS2 to ERG or ETV1 in prostate cancer tissues with outlier expression. By using fluorescence in situ hybridization, we demonstrated that 23 of 29 prostate cancer samples harbor rearrangements in ERG or ETV1. Cell line experiments suggest that the androgen-responsive promoter elements of TMPRSS2 mediate the overexpression of ETS family members in prostate cancer. These results have implications in the development of carcinomas and the molecular diagnosis and treatment of prostate cancer. PMID- 16254183 TI - Dating the growth of oceanic crust at a slow-spreading ridge. AB - Nineteen uranium-lead zircon ages of lower crustal gabbros from Atlantis Bank, Southwest Indian Ridge, constrain the growth and construction of oceanic crust at this slow-spreading midocean ridge. Approximately 75% of the gabbros accreted within error of the predicted seafloor magnetic age, whereas approximately 25% are significantly older. These anomalously old samples suggest either spatially varying stochastic intrusion at the ridge axis or, more likely, crystallization of older gabbros at depths of approximately 5 to 18 kilometers below the base of crust in the cold, axial lithosphere, which were uplifted and intruded by shallow level magmas during the creation of Atlantis Bank. PMID- 16254184 TI - The V-antigen of Yersinia forms a distinct structure at the tip of injectisome needles. AB - Many pathogenic bacteria use injectisomes to deliver effector proteins into host cells through type III secretion. Injectisomes consist of a basal body embedded in the bacterial membranes and a needle. In Yersinia, translocation of effectors requires the YopB and YopD proteins, which form a pore in the target cell membrane, and the LcrV protein, which assists the assembly of the pore. Here we report that LcrV forms a distinct structure at the tip of the needle, the tip complex. This unique localization of LcrV may explain its crucial role in the translocation process and its efficacy as the main protective antigen against plague. PMID- 16254185 TI - Neurogenesis in the hypothalamus of adult mice: potential role in energy balance. AB - Ciliary neurotrophic factor (CNTF) induces weight loss in obese rodents and humans, and for reasons that are not understood, its effects persist after the cessation of treatment. Here we demonstrate that centrally administered CNTF induces cell proliferation in feeding centers of the murine hypothalamus. Many of the newborn cells express neuronal markers and show functional phenotypes relevant for energy-balance control, including a capacity for leptin-induced phosphorylation of signal transducer and activator of transcription 3 (STAT3). Coadministration of the mitotic blocker cytosine-beta-d-arabinofuranoside (Ara-C) eliminates the proliferation of neural cells and abrogates the long-term, but not the short-term, effect of CNTF on body weight. These findings link the sustained effect of CNTF on energy balance to hypothalamic neurogenesis and suggest that regulated hypothalamic neurogenesis in adult mice may play a previously unappreciated role in physiology and disease. PMID- 16254186 TI - NPY/AgRP neurons are essential for feeding in adult mice but can be ablated in neonates. AB - Hypothalamic neurons that express neuropeptide Y (NPY) and agouti-related protein (AgRP) are thought to be critical regulators of feeding behavior and body weight. To determine whether NPY/AgRP neurons are essential in mice, we targeted the human diphtheria toxin receptor to the Agrp locus, which allows temporally controlled ablation of NPY/AgRP neurons to occur after an injection of diphtheria toxin. Neonatal ablation of NPY/AgRP neurons had minimal effects on feeding, whereas their ablation in adults caused rapid starvation. These results suggest that network-based compensatory mechanisms can develop after the ablation of NPY/AgRP neurons in neonates but do not readily occur when these neurons become essential in adults. PMID- 16254187 TI - The role of the BRCA1 tumor suppressor in DNA double-strand break repair. AB - The tumor suppressor gene BRCA1 was cloned in 1994 based on its linkage to early onset breast and ovarian cancer. Although the BRCA1 protein has been implicated in multiple cellular functions, the precise mechanism that determines its tumor suppressor activity is not defined. Currently, the emerging picture is that BRCA1 plays an important role in maintaining genomic integrity by protecting cells from double-strand breaks (DSB) that arise during DNA replication or after DNA damage. The DSB repair pathways available in mammalian cells are homologous recombination and nonhomologous end-joining. BRCA1 function seems to be regulated by specific phosphorylations in response to DNA damage and we will focus this review on the roles played by BRCA1 in DNA repair and cell cycle checkpoints. Finally, we will explore the idea that tumor suppression by BRCA1 depends on its control of DNA DSB repair, resulting in the promotion of error-free and the inhibition of error prone recombinational repair. PMID- 16254188 TI - EphA2 as a novel molecular marker and target in glioblastoma multiforme. AB - We investigated the presence of EphA2, and its ligand, ephrinA1, in glioblastoma multiforme (GBM), a malignant neoplasm of glial cells, and normal brain. We also initially examined the functional importance of the interaction between EphA2 and ephrinA1 in glioma cells. Expression and localization of EphA2 and ephrinA1 in human GBM and normal brain were examined using Western blotting, immunofluorescence, and immunohistochemistry. A functional role for EphA2 was investigated by assessing the activation status of the receptor and the effect of ephrinA1 on the anchorage-independent growth and invasiveness of GBM cells. We found EphA2 to be elevated in approximately 90% of GBM specimens and cell lines but not in normal brain, whereas ephrinA1 was present at consistently low levels in both GBM and normal brain. EphA2 was activated and phosphorylated by ephrinA1 in GBM cells. Furthermore, ephrinA1 induced a prominent, dose-dependent inhibitory effect on the anchorage-independent growth and invasiveness of GBM cells highly overexpressing EphA2, which was not seen in cells expressing low levels of the receptor. Thus, EphA2 is both specifically overexpressed in GBM and expressed differentially with respect to its ligand, ephrinA1, which may reflect on the oncogenic processes of malignant glioma cells. EphA2 seems to be functionally important in GBM cells and thus may play an important role in GBM pathogenesis. Hence, EphA2 represents a new marker and novel target for the development of molecular therapeutics against GBM. PMID- 16254189 TI - Fractionated low-dose radiation exposure leads to accumulation of DNA damage and profound alterations in DNA and histone methylation in the murine thymus. AB - Thymus, an important component of hematopoietic tissue, is a well-documented "target" of radiation carcinogenesis. Both acute and fractionated irradiation result in a high risk of leukemia and thymic lymphoma. However, the exact mechanisms underlying radiation-induced predisposition to leukemia and lymphoma are still unknown, and the contributions of genetic and epigenetic mechanisms in particular have yet to be defined. Global DNA hypomethylation is a well-known characteristic of cancer cells. Recent studies have also shown that tumor cells undergo prominent changes in histone methylation, particularly a substantial loss of trimethylation of histone H4-Lys20 and demethylation of genomic DNA. These losses are considered a universal marker of malignant transformation. In the present study, we investigated the effect of low-dose radiation exposure on the accumulation of DNA lesions and alterations of DNA methylation and histone H4 Lys20 trimethylation in the thymus tissue using an in vivo murine model. For the first time, we show that fractionated whole-body application of 0.5 Gy X-ray leads to decrease in histone H4-Lys20 trimethylation in the thymus. The loss of histone H4-Lys20 trimethylation was accompanied by a significant decrease in global DNA methylation as well as the accumulation of DNA damage as monitored by persistence of histone gammaH2AX foci in the thymus tissue of mice exposed to fractionated irradiation. Altered DNA methylation was associated with reduced expression of maintenance (DNMT1) and, to a lesser extent, de novo DNA methyltransferase DNMT3a in exposed animals. Expression of another de novo DNA methyltransferase DNMT3b was decreased only in males. Irradiation also resulted in approximately 20% reduction in the levels of methyl-binding proteins MeCP2 and MBD2. Our results show the involvement of epigenetic alterations in radiation induced responses in vivo. These changes may play a role in genome destabilization that ultimately leads to cancer. PMID- 16254190 TI - Induction of invasive mouse skin carcinomas in transgenic mice with mutations in both H-ras and p53. AB - Synergistic interaction between H-ras and p53 were systematically examined during skin tumorigenesis. Concurrent expression of an activated H-ras gene and a mutant p53 gene was accomplished by crossing p53(Val135/wt) mice with TG.AC mice. Topical application to wild-type mice with benzo(a)pyrene (BaP) alone produced approximately 26% skin tumor incidence, whereas BaP treatment of p53(wt/wt)Hras(TG.AC/wt), p53(Val135/wt)Hras(wt/wt), and p53(Val135/wt)Hras(TG.AC/wt) mice produced a 75%, 77%, and 100% incidence of skin tumors, respectively. An average of 0.33 tumor per mouse was observed in wild type (p53(wt/wt)Hras(wt/wt)) mice, whereas approximately 1.54, 1.96, and 3.08 tumors per mouse were seen in BaP-treated p53(wt/wt)Hras(TG.AC/wt), p53(Val135/wt)Hras(wt/wt), and p53(Val135/wt)Hras(TG.AC/wt) mice, respectively. The effects on total tumor volume were even more striking with 7-, 48-, and 588 fold increases in tumor volume compared with wild-type (p53(wt/wt)Hras(wt/wt)) in p53(wt/wt)Hras(TG.AC/wt), p53(Val135/wt)Hras(wt/wt), and p53(Val135/wt)Hras(TG.AC/wt) mice, respectively. Histopathologically, all tumors from p53(wt/wt)Hras(wt/wt) mice were either papillomas or well-differentiated squamous cell carcinomas, whereas the tumors in p53(wt/wt)Hras(TG.AC/wt), p53(Val135/wt)Hras(wt/wt), and p53(Val135/wt)Hras(TG.AC/wt) mice were principally squamous cell carcinomas with varying degree of invasiveness. Particularly, tumors in p53(Val135/wt)Hras(TG.AC/wt) mice exhibited the most rapid growth and the extreme form of tumor invasion. Microarray analysis revealed that dominant negative p53 (Val135) and activated H-ras affected several cellular processes involved in tumorigenesis possibly through its effects on apoptosis, cell cycle arrest, and Ras-mitogen-activated protein kinase pathways. The present study provides the first in vivo evidence that a germ line p53 mutation and activated H ras act synergistically to profoundly enhance tumor progression. PMID- 16254191 TI - A mechanism of COOH-terminal binding protein-mediated repression. AB - The E2F4 and E2F5 proteins specifically associate with the Rb-related p130 protein in quiescent cells to repress transcription of various genes encoding proteins important for cell growth. A series of reports has provided evidence that Rb-mediated repression involves both histone deacetylase (HDAC)-dependent and HDAC-independent events. Our previous results suggest that one such mechanism for Rb-mediated repression, independent of recruitment of HDAC, involves the recruitment of the COOH-terminal binding protein (CtBP) corepressor, a protein now recognized to play a widespread role in transcriptional repression. We now find that CtBP can interact with the histone acetyltransferase, cyclic AMP- responsive element--binding protein (CREB) binding protein, and inhibit its ability to acetylate histone. This inhibition is dependent on a NH2-terminal region of CtBP that is also required for transcription repression. These results thus suggest two complementary mechanisms for E2F/p130-mediated repression that have in common the control of histone acetylation at target promoters. PMID- 16254193 TI - Dipeptidyl-peptidase IV converts intact B-type natriuretic peptide into its des SerPro form. AB - BACKGROUND: Analysis of plasma B-type natriuretic peptide (BNP) has suggested the in vivo formation of a truncated form, BNP (3-32), also called des-SerPro-BNP. The objectives of this study were to investigate (a) whether BNP and other natriuretic peptides are truncated by dipeptidyl-peptidase IV (DPP IV/CD26; EC 3.4.14.5) and (b) whether this truncation affects the susceptibility to cleavage by neutral endopeptidase (NEP; EC 3.4.24.11). METHODS: Human BNP (1-32), A-type natriuretic peptide 1-28 (ANP 1-28), and related peptides were incubated with purified DPP IV and with human plasma. In addition, BNP (1-32), BNP (3-32), and ANP (1-28) were subjected to hydrolysis by NEP. Cleavage products were analyzed by mass spectrometry. RESULTS: BNP (1-32) was cleaved by purified DPP IV with a specificity constant of 0.37 x 10(6) L.mol(-1).s(-1). The DPP IV activity in EDTA plasma was able to truncate BNP (1-32) ex vivo. Addition of Vildagliptin, a specific DPP IV inhibitor, prevented this truncation in a concentration-dependent manner. Under in vitro circumstances in which ANP was hydrolyzed extensively, BNP (1-32) and BNP (3-32) were very resistant to NEP-mediated cleavage. CONCLUSIONS: DPP IV cleaves BNP (1-32) with an efficiency higher than or comparable to several known in vivo substrates of the enzyme. Even after loss of the amino-terminal dipeptide, BNP remains highly resistant to cleavage by NEP. PMID- 16254192 TI - CCAAT/enhancer binding protein alpha (C/EBPalpha) and C/EBPalpha myeloid oncoproteins induce bcl-2 via interaction of their basic regions with nuclear factor-kappaB p50. AB - The CEBPA gene is mutated in 10% of acute myeloid leukemia (AML) cases. We find that CEBPA and Bcl-2 RNA levels correlate highly in low-risk human AMLs, suggesting that inhibition of apoptosis via induction of bcl-2 by CCAAT/enhancer binding protein alpha (C/EBPalpha) or its mutant variants contributes to transformation. C/EBPalphap30, lacking a NH2-terminal transactivation domain, or C/EBPalphaLZ, carrying in-frame mutations in the leucine zipper that prevent DNA binding, induced bcl-2 in hematopoietic cell lines, and C/EBPalpha induced bcl-2 in normal murine myeloid progenitors and in the splenocytes of H2K-C/EBPalpha-Emu transgenic mice. C/EBPalpha protected Ba/F3 cells from apoptosis on interleukin-3 withdrawal but not if bcl-2 was knocked down. Remarkably, C/EBPalphaLZ oncoproteins activated the bcl-2 P2 promoter despite lack of DNA binding, and C/EBPalphap30 also activated the promoter. C/EBPalpha and the C/EBPalpha oncoproteins cooperated with nuclear factor-kappaB (NF-kappaB) p50, but not p65, to induce bcl-2 transcription. Endogenous C/EBPalpha preferentially coimmunoprecipitated with p50 versus p65 in myeloid cell extracts. Mutation of residues 297 to 302 in the C/EBPalpha basic region prevented induction of endogenous bcl-2 or the bcl-2 promoter and interaction with p50 but not p65. These findings suggest that C/EBPalpha or its mutant variants tether to a subset of NF-kappaB target genes, including Bcl-2, via p50 to facilitate gene activation and offer an explanation for preferential in-frame rather than out-of-frame mutation of the leucine zipper with sparing of the basic region in C/EBPalphaLZ oncoproteins. Targeting interaction between C/EBPalpha basic region and NF-kappaB p50 may contribute to the therapy of AML and other malignancies expressing C/EBPs. PMID- 16254194 TI - Application of proteomic technology in identifying pancreatic secretory trypsin inhibitor variants in urine of patients with pancreatitis. AB - BACKGROUND: Although the analysis of genetic variability has traditionally been performed with molecular genetic techniques, the development of proteomic technology has raised the possibility of analyzing genetic variants at the protein level. This method provides additional information about posttranslational modifications and differences in expression. We used mass spectrometry to characterize 3 variants of the peptide encoded by the serine protease inhibitor Kazal type 1 (SPINK1) gene, pancreatic secretory trypsin inhibitor (PSTI). A genetic variant of PSTI, N34S, is associated with the development of pancreatitis. METHODS: We used a quadrupole/time-of-flight hybrid mass spectrometer equipped with an electrospray ionization source to analyze the molecular identity of PSTI purified from the urine of 12 patients with pancreatitis and from 3 controls. We also developed a rapid small-scale capture procedure to isolate and analyze PSTI from small volumes of urine. RESULTS: The mutations responsible for mass shifts of different PSTI variants could be verified. We observed differences in the expression of different variants as well as a novel proteolytic fragment of PSTI. Small-scale magnetic bead-mediated immunoaffinity chromatography PSTI enabled easy and rapid purification from small urine volumes, facilitating mass spectrometric analysis with adequate sensitivity. CONCLUSIONS: Pancreatitis-related PSTI variants occurring at nanomolar concentrations in urine can be detected and quantified by immunoaffinity purification and mass spectrometry. In addition, the N34S variant occurs at higher concentrations than the wild type. This finding casts new light on the possible role of PSTI as a cause of hereditary pancreatitis. PMID- 16254195 TI - Population differences in the polyalanine domain and 6 new mutations in HLXB9 in patients with Currarino syndrome. AB - BACKGROUND: The combination of partial absence of the sacrum, anorectal anomalies, and presacral mass constitutes Currarino syndrome (CS), which is associated with mutations in HLXB9. METHODS: We analyzed 5 CS families and 6 sporadic cases for HLXB9 mutations by direct sequencing. Potentially pathologic expansions of HLXB9 GCC repeats were analyzed in patients, 4 general populations [Chinese, Japanese, Yoruba, and Centre du Etude Polymorphisme Human (CEPH)] from the HapMap project, and 145 healthy Chinese. RESULTS: We identified 6 novel mutations affecting highly conserved residues (Ser185X, Trp215X, Ala26fs, Ala75fs, Met1Ile, and Arg273Cys). GCC allele and genotype distributions showed marked statistically significant differences. (GCC)11 was the most common allele overall; its frequency ranged from 90% in CEPH to 68% in Yoruba and 50% in Chinese and Japanese populations. (GCC)9 was almost as common as (GCC)11 in Chinese and Japanese populations, whereas its frequency was <10% in Yoruba and CEPH populations. The Yoruba population had the highest frequency of the largest alleles [(GCC)12 and (GCC)13], which were almost absent in the other groups. CONCLUSIONS: Lack of HLXB9 mutations in some patients and the presence of variable phenotypes suggest DNA alterations in HLXB9 noncoding regions and/or in other genes encoding HLXB9 regulatory molecules or protein partners. If HLXB9, like other homeobox genes, has a threshold beyond which triplet expansions are pathologic, those populations enriched with larger alleles would be at a higher risk. The data illustrate the importance of ethnicity adjustment if these polymorphic markers are to be used in association studies. PMID- 16254196 TI - Thyroid function and prevalence of anti-thyroperoxidase antibodies in a population with borderline sufficient iodine intake: influences of age and sex. AB - BACKGROUND: We present a large European population-based study of thyroid function, performed in a population with longstanding borderline sufficient iodine intake. METHODS: The Nijmegen Biomedical Study is a population-based survey conducted in the eastern part of The Netherlands. Randomly selected inhabitants received a postal questionnaire on lifestyle and medical history, which was filled out by 9371 individuals (41.7%). We measured serum thyroid stimulating hormone (TSH), free thyroxine (FT4), and anti-thyroperoxidase antibodies (TPOAbs) in 6434 responders. A reference population of 5167 individuals was selected by excluding those at risk for thyroid disease. RESULTS: Overt thyrotoxicosis was found in 0.4% of the total population and subclinical thyrotoxicosis in 0.8%. Overt hypothyroidism was found in 0.4% and subclinical hypothyroidism in 4.0%. In individuals older than 60 years, mean FT4 concentrations increased with age. Mean TSH decreased with age, from 1.46 mIU/L at 18-24 years to 1.07 mIU/L after 85 years. The mean TSH in the total population did not differ from the mean TSH in the reference population; the exclusion of those at risk for thyroid disease, however, lowered the upper limit of the TSH reference interval considerably. In the total population, 8.6% of males and 18.5% of females had positive TPOAbs. The presence of TPOAbs was associated with abnormally high and low TSH concentrations. CONCLUSION: In inhabitants of the eastern part of The Netherlands, serum TSH gradually decreases with age, whereas after age 60, serum FT4 increases, possibly because of the development of thyroid autonomy after longstanding borderline sufficient iodine intake. PMID- 16254197 TI - Interaction of the -308G/A promoter polymorphism of the tumor necrosis factor alpha gene with single-nucleotide polymorphism 45 of the adiponectin gene: effect on serum adiponectin concentrations in a Spanish population. AB - BACKGROUND: We investigated whether interactions of the -308G/A polymorphism in the promoter region of the tumor necrosis factor-alpha (TNF-alpha) gene with single-nucleotide polymorphisms (SNPs) 45 and 276 of the adiponectin gene are associated with circulating adiponectin and soluble TNF-alpha receptor 2 (sTNFR2) concentrations in a Spanish population. METHODS: We performed anthropometric and physiologic measurements in 809 unrelated participants recruited with a simple random sampling approach from respondents to a cross-sectional population-based epidemiologic survey in the province of Segovia in central Spain (Castille). RESULTS: The 2-h postload glucose and serum insulin concentrations were higher in -308A allele carriers than in -308G/G individuals homozygous for the TNF-alpha gene. Plasma concentrations of sTNFR2 were positively correlated with body mass index, waist-to-hip ratio, and sagittal abdominal diameter among individuals with type 2 diabetes. Individuals with type 2 diabetes and the -308A allele had higher sTNFR2 and lower adiponectin concentrations than -308G homozygotes. Moreover, individuals carrying both the TNF-alpha -308A allele and the G allele of SNP 45 in the adiponectin gene had the highest prevalence of impaired glucose tolerance (adjusted odds ratio, 1.26; 95% confidence interval, 1.01-1.56; P = 0.038) and had lower adiponectin concentrations (beta = -0.090; P = 0.005) than individuals without these genotypes. CONCLUSIONS: Our findings are the first to indicate that a higher incidence of impaired glucose tolerance and low circulating adiponectin concentration may be associated with interaction between the -308G/A promoter polymorphism of the TNF-alpha gene and SNP 45 in the adiponectin gene. PMID- 16254198 TI - Distinct cellular loci for the ABCA1-dependent and ABCA1-independent lipid efflux mediated by endogenous apolipoprotein E expression. AB - OBJECTIVE: Macrophage expression of both apolipoprotein E (apoE) and ABCA1 have been shown to modulate lipid efflux from these cells and to play an important atheroprotective role in vivo. We evaluated the relationship between apoE and ABCA1 for regulating cellular sterol efflux. METHODS AND RESULTS: ApoE-mediated, but ABCA1-independent, lipid efflux was demonstrated in 3 model systems. First, adenoviral-mediated expression of apoE in dermal fibroblasts isolated from ABCA1( /-) mice significantly increased both sterol and phospholipid efflux. Second, expression of human apoE in a macrophage cell line increased sterol efflux, and this increment in efflux was not reduced by suppressing ABCA1 expression. Third, reduction of apoE expression using an apoE small interfering RNA significantly reduced sterol efflux from ABCA1(-/-) mouse peritoneal macrophages. ApoE mediated, but ABCA1-independent, lipid efflux could be differentiated from lipid efflux that was dependent on the extracellular accumulation of secreted apoE, because exogenous cell-derived apoE stimulated efflux only from cells expressing ABCA1. Sterol efflux was usually highest in cells expressing both ABCA1 and apoE, likely representing a summation of the ABCA1-dependent and -independent pathways for apoE-mediated sterol efflux. CONCLUSIONS: ABCA1 expression is required for apoE-mediated efflux when endogenously synthesized apoE accumulates extracellularly. Our results, however, establish the existence of an ABCA1 independent pathway for lipid efflux that requires the intracellular synthesis and/or transport of apoE. PMID- 16254199 TI - Low-density lipoprotein cholesterol determines oxidative stress and endothelial dysfunction in saphenous veins from patients with coronary artery disease. AB - OBJECTIVE: There is evidence for a relationship between endothelial dysfunction and cardiovascular disease, but a causative role for oxidative stress remains to be determined. METHODS AND RESULTS: We studied 188 patients with severe coronary artery disease (CAD), of whom 51 were age and sex matched with 51 healthy controls undergoing varicose vein surgery. Relaxation of saphenous vein to calcium ionophore, apocynin, and allopurinol was studied together with the markers of oxidative stress, total antioxidant capacity and reduced/oxidized glutathione ratio. Vascular superoxide levels were measured using lucigenin chemiluminescence and hydroethidine. Relaxation to calcium ionophore was decreased in CAD compared with control patients (maximum relaxation 26+/-2% versus 60+/-1%; P<0.001). Total superoxide production was increased (0.89+/-0.09 versus 0.56+/-0.06 nmol/mg per min; P=0.008), whereas superoxide inhibition with apocynin or allopurinol had a greater effect on vasorelaxation in CAD patients. Low-density lipoprotein (LDL) cholesterol predicted relaxation to calcium ionophore (P<0.001) and oxidative stress markers (P<0.001) in CAD patients. CONCLUSIONS: Endothelial dysfunction is associated with raised levels of superoxide and biomarkers of oxidative stress in saphenous veins from CAD patients. LDL cholesterol is a major determinant of endothelial dysfunction and oxidative stress in these patients. These results support intensive LDL cholesterol-lowering therapy as suggested by recent clinical trials. PMID- 16254200 TI - Role of blood cell-associated AT1 receptors in the microvascular responses to hypercholesterolemia. AB - OBJECTIVE: Hypercholesterolemia elicits a proinflammatory and prothrombogenic phenotype in the microvasculature that is characterized by activation and adhesion of blood cells. The angiotensin II receptor-1 antagonist Losartan prevents the induction of these responses. The objective of this study was to determine the relative contributions of blood cell-associated versus endothelium associated AT1a-R to these hypercholesterolemia-induced microvascular alterations. METHODS AND RESULTS: Leukocyte adhesion and emigration and platelet adhesion were quantified by intravital microscopy in postcapillary venules. C57Bl/6 mice were placed on a normal (ND) or high-cholesterol (HCD) diet for 2 weeks. AT1a-R bone marrow chimeras that express AT1a-R on the vessel wall but not blood cells and AT1a-R knockouts were placed on HCD. Venular shear rate was comparable in all groups. Platelet and leukocyte adhesion and leukocyte emigration were significantly increased in HCD mice versus ND. Leukocyte recruitment was significantly reduced in the HCD-AT1a-R bone marrow chimera group, whereas platelet adhesion remained at HCD levels. However, in HCD-AT1a-R knockout mice, platelet and leukocyte adhesion were reduced to ND levels. CONCLUSIONS: These data indicate that the platelet-vessel wall adhesion elicited by hypercholesterolemia is mediated by AT1a-R engagement on the endothelial cell rather than the platelet, whereas leukocyte recruitment is mediated by blood cell associated AT1a-R. PMID- 16254201 TI - What does it take to make the perfect clot? AB - The coagulation process has been conceptualized as being primarily dependent on adequate levels of the coagulation proteins. This concept was based on the clear relationship between the bleeding tendency and factor levels in hemophilia. The field is now evolving toward conceptualizing coagulation as being actively regulated by the specialized cellular components of the process. Rather than conceiving coagulation as only a "cascade" of proteolytic reactions, the coagulation reactions occur as overlapping steps on cell surfaces. Components of the old "extrinsic'" and "intrinsic" pathways of coagulation can be thought of as participating in the initiation and propagation of coagulation reactions, respectively. Thus, these pathways are not redundant as they are portrayed in the cascade model, but play distinct and complementary roles. Our understanding of how specific cellular features control the processes of hemostasis and thrombosis is developing rapidly. This review discusses some aspects of the cellular control of coagulation. PMID- 16254202 TI - Platelet nitric oxide responsiveness: a novel prognostic marker in acute coronary syndromes. AB - OBJECTIVE: Nitric oxide (NO) is critically important in the regulation of vascular tone and the inhibition of platelet aggregation. We have shown previously that patients with acute coronary syndromes (ACS) or stable angina pectoris have impaired platelet responses to NO donors when compared with normal subjects. We tested the hypotheses that platelet hyporesponsiveness to NO is a predictor of (1) cardiovascular readmission and/or death and (2) all-cause mortality in patients with ACS (unstable angina pectoris or non-Q-wave myocardial infarction). METHODS AND RESULTS: Patients (n=51) with ACS had evaluation of platelet aggregation within 24 hours of coronary care unit admission using impedance aggregometry. Patients were categorized as having "normal" (> or =32% inhibition of ADP-induced aggregation with the NO donor sodium nitroprusside; 10 micromol/L; n=18) or "impaired" (<32% inhibition of ADP-induced aggregation; n=33) NO responses. We then compared the incidence of cardiovascular readmission and death during a median of 7 years of follow-up in these 2 groups. Using a Cox proportional hazards model adjusting for age, sex, index event, postdischarge medical treatment, revascularization status, left ventricular systolic dysfunction, concurrent disease states, and cardiac risk factors, impaired NO responsiveness was associated with an increased risk of the combination of cardiovascular readmission and/or death (relative risk, 2.7; 95% CI, 1.03 to 7.10; P=0.041) and all-cause mortality (relative risk, 6.3; 95% CI, 1.09 to 36.7; P=0.033). CONCLUSIONS: Impaired platelet NO responsiveness is a novel, independent predictor of increased mortality and cardiovascular morbidity in patients with high-risk ACS. PMID- 16254203 TI - Stimulation of liver-directed cholesterol flux in mice by novel N acetylgalactosamine-terminated glycolipids with high affinity for the asialoglycoprotein receptor. AB - OBJECTIVE: Interventions that promote liver-directed cholesterol flux can suppress atherosclerosis, as demonstrated for scavenger receptor-BI overexpression in hypercholesterolemic mice. In analogy, we speculate that increasing lipoprotein flux to the liver via the asialoglycoprotein receptor (ASGPr) may be of therapeutic value in hypercholesterolemia. METHODS AND RESULTS: A bifunctional glycolipid (LCO-Tyr-GalNAc3) with a high-nanomolar affinity for the ASGPr (inhibition constant 2.1+/-0.2 nmol/L) was synthesized that showed rapid association with lipoproteins on incubation with serum. Prior incubation of LCO-Tyr-GalNAc3 with radiolabeled low-density lipoprotein or high-density lipoprotein (0.5 microg/microg of protein) resulted in a dramatic induction of the liver uptake of these lipoproteins when injected intravenously into mice (70+/-3% and 78+/-1%, respectively, of the injected dose at 10 minutes of low density lipoprotein and high-density lipoprotein), as mediated by the ASGPr on hepatocytes. Intravenously injected LCO-Tyr-GalNAc3 quantitatively incorporated into serum lipoproteins and evoked a strong and persistent (> or =48 hour) cholesterol-lowering effect in normolipidemic mice (37+/-2% at 6 hours) and hyperlipidemic apoE(-/-) mice (32+/-2% at 6 hours). The glycolipid was also effective on subcutaneous administration. CONCLUSIONS: LCO-Tyr-GalNAc3 is very effective in promoting cholesterol uptake by hepatocytes and, thus, may be a promising alternative for the treatment of those hyperlipidemic patients who do not respond sufficiently to conventional cholesterol-lowering therapies. PMID- 16254204 TI - Chemokines in children with heterozygous familiar hypercholesterolemia: selective upregulation of RANTES. AB - OBJECTIVE: Increasing data support the involvement of chemokines in atherogenesis. However, although several studies have shown increased chemokine levels in adult patients, the literature is virtually devoid of data on chemokines in children with hypercholesterolemia. METHODS AND RESULTS: We examined the gene expression of chemokines in peripheral blood mononuclear cells (PBMCs) from clinically healthy children with and without heterozygous familial hypercholesterolemia (FH). Our main findings were: (1) compared with healthy controls, PBMCs from FH children showed significantly higher mRNA levels of RANTES, but not of the other examined chemokines; (2) an opposite pattern was seen in adult FH subjects, with markedly enhanced expression of macrophage inflammatory peptide-1alpha, but not of RANTES; (3) this increased gene expression of RANTES in PBMCs from FH children seemed to reflect enhanced RANTES expression in monocytes but not in T cells; (4) FH children also had raised serum levels of neopterin, additionally suggesting monocyte/macrophage activation in these children; and (5) PBMCs from both FH children and controls showed enhanced release of interleukin 8 on RANTES stimulation in vitro. CONCLUSIONS: Our findings support a role of inflammation also in the early stages of atherogenesis possibly involving monocyte-derived RANTES as an important mediator. PMID- 16254205 TI - Cholesteryl esters of aggregated LDL are internalized by selective uptake in human vascular smooth muscle cells. AB - OBJECTIVE: Low-density lipoprotein (LDL) receptor-related protein (LRP1) mediates the internalization of aggregated LDL (agLDL)-LDL trapped in the arterial intima bound to proteoglycans-into human vascular smooth muscle cells (VSMC). LRP1 mediated agLDL uptake induces high-intracellular cholesteryl ester (CE) accumulation. The aim of this study was to characterize the mechanism of agLDL internalization in human VSMC. METHODS AND RESULTS: The lipidic component of LDL was labeled with [3H] and the apolipoprotein component with [(125)I]. We found that >90% of intracellular CE derived from agLDL uptake was not associated with apoB100 degradation but was selectively taken up from agLDL. The inhibition of LRP1 expression by small interfering RNA treatment led to a decrease of 80+/ 0.05% in agLDL-CE selective uptake. AgLDL induced intracellular CE accumulation without a concomitant CE synthesis. Cytosolic and cytoskeletal proteins were not required for CE transport. Electron and confocal microscopy experiments indicate that CE derived from agLDL accumulated in adipophilin-stained lipid droplets that were not removable by high-density lipoprotein. CONCLUSIONS: Taken together, these results demonstrate that LRP1 mediates the selective uptake of CE from agLDL and that CE derived from agLDL is not intracellularly processed but stored in lipid droplets in human VSMC. PMID- 16254206 TI - Beta-catenin overexpression augments angiogenesis and skeletal muscle regeneration through dual mechanism of vascular endothelial growth factor mediated endothelial cell proliferation and progenitor cell mobilization. AB - OBJECTIVE: Beta-catenin plays a critical role in directing cell fate during embryogenesis, and uncontrollable activation leads to cancers, suggesting its importance in cell survival and proliferation. However, little is known regarding its role in endothelial cell (EC) and skeletal muscle proliferation and progenitor cell mobilization. METHODS AND RESULTS: Beta-catenin enhanced ECs proliferation, protected ECs from apoptosis, and increased the capillary forming capabilities, which was completely blocked by inhibition of its nuclear translocation. In addition, the increased proliferation by beta-catenin was associated with increased expression of cyclin E2. In skeletal myocytes, beta catenin overexpression increased proliferation with cyclin D1 expression, decreased apoptosis, and induced hypertrophy. Furthermore, beta-catenin induced the expression of vascular endothelial growth factor (VEGF) in skeletal myocytes, resulting in EC proliferation. In a mouse hindlimb ischemia model, beta-catenin significantly increased recovery of blood perfusion, capillary density along with enhanced VEGF expression, and the number of proliferating ECs and myocytes. Local delivery of beta-catenin also promoted angiogenic progenitor cell mobilization and increased the number of satellite cells. CONCLUSIONS: Beta-catenin may be an important modulator of angiogenesis and myocyte regeneration not only by directly enhancing proliferation and survival of ECs and skeletal myocytes but also by inducing VEGF expression and promoting angiogenic progenitor cell mobilization and muscle progenitor cell activation. PMID- 16254207 TI - The glycoprotein VI-phospholipase Cgamma2 signaling pathway controls thrombus formation induced by collagen and tissue factor in vitro and in vivo. AB - OBJECTIVE: Both collagen and tissue factor can be initiating factors in thrombus formation. We investigated the signaling pathway of collagen-induced platelet activation in interaction with tissue factor-triggered coagulation during the thrombus-forming process. METHODS AND RESULTS: In murine blood flowing over collagen, platelet exposure of phosphatidylserine and procoagulant activity, but not adhesion, completely relied on each of the following signaling modules: glycoprotein VI (GPVI), FcR gamma-chain, Src kinases, adaptor protein LAT, and phospholipase Cgamma2 (PLCgamma2). On flow in the presence of tissue factor, these signaling components were essential for platelet aggregation and greatly enhanced fibrin clot formation. Collagen-stimulated thrombin generation relied on the presence and activity of GPVI, FcR gamma-chain, Src kinase, LAT, and PLCgamma2. The physiological importance of this GPVI pathway was shown in a FeCl3 induced in vivo murine thrombosis model. In both venules and arterioles, signaling through GPVI, FcR gamma-chain, and Src kinases enhanced the formation of phosphatidylserine-exposing and fibrin-rich thrombi. CONCLUSIONS: The GPVI PLCgamma2 activation pathway regulates collagen-dependent coagulation in venous and arterial thrombus formation. PMID- 16254208 TI - Expression of neutrophil gelatinase-associated lipocalin in atherosclerosis and myocardial infarction. AB - OBJECTIVE: Neutrophil gelatinase-associated lipocalin (NGAL) modulates the activity of matrix metalloproteinase (MMP) 9, an important mediator of vascular remodeling and plaque instability in atherosclerosis. This study aimed to analyze the expression of NGAL in atherosclerotic plaques and myocardial infarction (MI). METHODS AND RESULTS: Atherosclerotic apolipoprotein E (apoE)(-/-) x low-density lipoprotein receptor (LDLR)(-/-) and C57BL/6J control mice were exposed to brief hypoxic stress (10 minutes of 10% oxygen). Expression of the mouse NGAL homolog (24p3) and MMP-9 was analyzed 48 hours later by quantitative RT-PCR, immunohistochemistry, and zymography. Hypoxic stress increased NGAL/24p3 mRNA in the cardiac vasculature. NGAL/24p3 was also increased in atherosclerotic plaques of apolipoprotein E(-/-) x LDLR(-/-) mice compared with C57BL/6J mice. Mice developing MI exhibited the highest plaque mRNA expression of NGAL/24p3 and MMP 9. Zymography revealed strong proteolytic activity in areas rich in 24p3 and MMP 9 protein. Immunohistochemistry performed on human carotid endarterectomy specimens and control tissue from the internal mammary artery showed colocalization of MMP-9 and NGAL with macrophages in the atherosclerotic plaques. CONCLUSIONS: NGAL/24p3 is increased in atherosclerotic plaques and MI. Colocalization with MMP-9 in areas with high-proteolytic activity suggests a role for NGAL/24p3 in modulating the MMP-9-mediated remodeling of plaques and infarcted hearts. PMID- 16254209 TI - Glucocorticoid receptor regulates ATP-binding cassette transporter-A1 expression and apolipoprotein-mediated cholesterol efflux from macrophages. AB - OBJECTIVE: The ATP-binding cassette transporter-A1 (ABCA1) regulates cholesterol efflux from cells and is involved in high-density lipoprotein metabolism and atherogenesis. The objective of this study was to investigate the effect of dexamethasone (Dex) and other glucocorticoid receptor (GR) ligands on apolipoprotein AI-mediated cholesterol efflux from macrophages and ABCA1 expression in them. METHODS AND RESULTS: Dex, a GR agonist, decreased ABCA1 mRNA levels in a dose- and time-dependent fashion, and RU486, a GR antagonist, reversed the inhibitory effect of Dex. The effects of Dex and RU486 on ABCA1 protein levels and apolipoprotein AI-mediated cholesterol efflux from the macrophages were consistent with these changes in mRNA levels. Transfected RAW264.7, together with a human ABCA1 promoter-luciferase construct, inhibited transcriptional activity by Dex and overexpression of human GR. Transrepression by GR was not mediated by liver X receptor (LXR), because there were no differences in the effects of the GR ligands on promoter activity between a reporter construct with mutations at the LXR binding site and one without the mutations, and no changes were brought about in ABCG1 and ABCG4 expression by GR ligands. CONCLUSIONS: Our results showed that GR ligands affected ABCA1 expression and cholesterol efflux from macrophages, which are regulated by GR through a LXR-independent mechanism. PMID- 16254210 TI - Nicotine induces proinflammatory responses in macrophages and the aorta leading to acceleration of atherosclerosis in low-density lipoprotein receptor(-/-) mice. AB - OBJECTIVE: We investigated the molecular mechanism of nicotine-accelerated atherosclerosis in a hyperlipidemic low-density lipoprotein receptor(-/-) mouse model. METHODS AND RESULTS: Low-density lipoprotein receptor(-/-) mice received time-release nicotine or placebo pellets for 90 days. Aortic lesion size was 2.5 times larger in nicotine-treated than in placebo-treated mice (P<0.001). A mild increase in lipids was seen in treated mice. We quantified 18 different serum cytokines and found a significant increase of tumor necrosis factor alpha, interleukin 1beta, and keratinocyte-derived chemokine in nicotine-treated mice. Among 107 nuclear factor kappaB (NF-kappaB) target genes screened from the aorta, we found that nicotine treatment upregulated only 4 atherogenic genes including vascular adhesion molecule 1 and cyclooxygenase 2 on day 60 and platelet-derived growth factor B and platelet 12-lipoxygenase on day 90. At the cellular level, nicotine induced tumor necrosis factor alpha and inducible nitric oxide synthase expression in RAW264.7 cells via the nicotinic acetylcholine receptors. Induction was confirmed in peritoneal macrophages isolated from nicotine-treated mice. Finally, we showed that preconditioned medium from nicotine-treated RAW264.7 cells activated NF-kappaB in human smooth muscle cells and vascular endothelial cells as evidenced by nuclear localization and electromobility shift assay. CONCLUSIONS: Chronic nicotine exposure augments atherosclerosis by enhancing the production of proinflammatory cytokines by macrophages, which, in turn, activate atherogenic NF-kappaB target genes in the aortic lesions. PMID- 16254211 TI - Nuclear RNA foci in the heart in myotonic dystrophy. AB - The disease mechanism underlying myotonic dystrophy type 1 (DM1) pathogenesis in skeletal muscle may involve sequestration of RNA binding proteins in nuclear foci of expanded poly(CUG) RNA. Here we report evidence for a parallel mechanism in the heart. Accumulation of expanded poly(CUG) RNA in nuclear foci is associated with sequestration of muscleblind proteins and abnormal regulation of alternative splicing in DM1 cardiac muscle. A toxic effect of RNA with an expanded repeat may contribute to cardiac disease in DM1. PMID- 16254212 TI - Activation of the endothelial store-operated ISOC Ca2+ channel requires interaction of protein 4.1 with TRPC4. AB - Store-operated calcium (SOC) entry represents the principal Ca2+ entry pathway into nonexcitable cells. Despite intensive investigation, mechanisms underlying activation of SOC entry have remained elusive. The endothelial ISOC channel is a Ca2+-selective SOC entry channel to which the transient receptor potential (TRP) proteins TRPC1 and TRPC4 contribute subunits. Activation of ISOC is specifically regulated by the spectrin-actin membrane skeleton; however, the nature of coupling between the ISOC channel and membrane skeleton is unknown. Here we demonstrate that protein 4.1 is an essential component of the ISOC channel gating mechanism. Protein 4.1 interacts with TRPC4 and the membrane skeleton. Deletion of the protein 4.1 binding domain on TRPC4 or peptide competition to the protein 4.1 binding domain prevents ISOC activation. These findings reveal that interaction of protein 4.1 with TRPC4 is required for activation of the endothelial ISOC channel. PMID- 16254213 TI - Impaired CXCR4 signaling contributes to the reduced neovascularization capacity of endothelial progenitor cells from patients with coronary artery disease. AB - Transplantation of bone marrow cells as well as circulating endothelial progenitor cells (EPC) enhances neovascularization after ischemia. The chemokine receptor CXCR4 is essential for migration and homing of hematopoietic stem cells. Therefore, we investigated the role of CXCR4 and its downstream signaling cascade for the angiogenic capacity of cultured human EPC. Ex vivo, differentiated EPC derived from peripheral blood abundantly expressed CXCR4. Incubation of EPC from healthy volunteers with neutralizing antibodies against CXCR4 profoundly inhibited vascular endothelial growth factor- and stromal-derived factor-1 induced migration as well as EPC-induced angiogenesis in an ex vivo assay. Preincubation of transplanted EPC with CXCR4 antibody reduced EPC incorporation and impaired blood-flow recovery in ischemic hindlimbs of nude mice (57+/-4% of normal perfusion versus untreated EPC: 80+/-11%, P<0.001). Bone marrow mononuclear cells (BM-MNC) or EPC of heterozygous CXCR4(+/-) mice displayed reduced CXCR4 expression and disclosed impaired in vivo capacity to enhance recovery of ischemic blood flow in nude mice (blood flow 27+/-11% versus 66+/-25% using wild-type cells, P<0.01). Importantly, impaired blood flow in ischemic CXCR4(+/-) mice was rescued by injection of wild-type BM-MNC. Next, we investigated the role of CXCR4 for functional capacities of EPC from patients with coronary artery disease (CAD). Surface expression of CXCR4 was similar in EPC from patients with CAD compared with healthy controls. However, basal Janus kinase (JAK)-2 phosphorylation was significantly reduced and less responsive to stromal-derived factor-1 in EPC from patients with CAD compared with healthy volunteers, indicating that CXCR4-mediated JAK-2 signaling is dysregulated in EPC from patients with CAD. The CXCR4 receptor signaling profoundly modulates the angiogenic activity and homing capacity of cultured human EPC. Disturbance of CXCR4 signaling, as demonstrated by reduced JAK-2 phosphorylation, may contribute to functional impairment of EPC from patients with CAD. Stimulating CXCR4 signaling might improve functional properties of EPC and may rescue impaired neovascularization capacity of EPC derived from patients with CAD. PMID- 16254215 TI - Targeted uptake-1 carrier to rescue the failing heart. PMID- 16254216 TI - NAD(P)H oxidases and TGF-beta-induced cardiac fibroblast differentiation: Nox-4 gets Smad. PMID- 16254214 TI - Proinflammatory effects of bacterial recombinant human C-reactive protein are caused by contamination with bacterial products, not by C-reactive protein itself. AB - Intravenous administration to human volunteers of a commercial preparation of recombinant human C-reactive protein (CRP) produced in Escherichia coli was recently reported in this journal to induce an acute phase response of serum amyloid A protein (SAA) and of CRP itself, and to activate the coagulation system. The authors concluded that CRP is probably a mediator of atherothrombotic disease. Here we confirm that this recombinant CRP preparation was proinflammatory both for mouse macrophages in vitro and for mice in vivo, but show that pure natural human CRP had no such activity. Furthermore mice transgenic for human CRP, and expressing it throughout their lives, maintained normal concentrations of their most sensitive endogenous acute phase reactants, SAA and serum amyloid P component (SAP). The patterns of in vitro cytokine induction and of in vivo acute phase stimulation by the recombinant CRP preparation were consistent with contamination by bacterial products, and there was 46.6 EU of apparent endotoxin activity per mg of CRP in the bacterial product, compared with 0.9 EU per mg of our isolated natural human CRP preparation. The absence of any proinflammatory activity in natural CRP for macrophages or healthy mice strongly suggests that the in vivo effects of the recombinant preparation observed in humans were attributable to proinflammatory bacterial products and not human CRP. PMID- 16254217 TI - Recent developments in vascular endothelial cell transient receptor potential channels. AB - Among the 28 identified and unique mammalian TRP (transient receptor potential) channel isoforms, at least 19 are expressed in vascular endothelial cells. These channels appear to participate in a diverse range of vascular functions, including control of vascular tone, regulation of vascular permeability, mechanosensing, secretion, angiogenesis, endothelial cell proliferation, and endothelial cell apoptosis and death. Malfunction of these channels may result in disorders of the human cardiovascular system. All TRP channels, except for TRPM4 and TRPM5, are cation channels that allow Ca2+ influx. However, there is a daunting diversity in the mode of activation and regulation in each case. Specific TRP channels may be activated by different stimuli such as vasoactive agents, oxidative stress, mechanical stimuli, and heat. TRP channels may then transform these stimuli into changes in the cytosolic Ca2+, which are eventually coupled to various vascular responses. Evidence has been provided to suggest the involvement of at least the following TRP channels in vascular function: TRPC1, TRPC4, TRPC6, and TRPV1 in the control of vascular permeability; TRPC4, TRPV1, and TRPV4 in the regulation of vascular tone; TRPC4 in hypoxia-induced vascular remodeling; and TRPC3, TRPC4, and TRPM2 in oxidative stress-induced responses. However, in spite of the large body of data available, the functional role of many endothelial TRP channels is still poorly understood. Elucidating the mechanisms regulating the different endothelial TRP channels, and the associated development of drugs selectively to target the different isoforms, as a means to treat cardiovascular disease should, therefore, be a high priority. PMID- 16254218 TI - Risk of myocardial infarction and vascular death after transient ischemic attack and ischemic stroke: a systematic review and meta-analysis. AB - BACKGROUND: Whether stroke patients should be investigated for asymptomatic coronary artery disease remains matter of debate. Absolute risks of myocardial infarction (MI) and vascular death after a stroke have not been accurately assessed. We performed a systematic review and a meta-analysis to determine the risk of MI and nonstroke vascular death after transient ischemic attack (TIA) and ischemic stroke. Cohort studies of TIA or ischemic stroke patients were included if they were published between 1980 and March 2005, reported risk of MI and nonstroke vascular death, enrolled >100 patients, and had at least 1 year of follow-up. We included 39 studies in a total of 65,996 patients with mean follow up of 3.5 years. Two reviewers independently carried out data extraction using a standardized form. Absolute annual risks were estimated through weighted meta regressions with a random effect. To test the predictions of expected event rates derived from our analysis, we used individual patient data. SUMMARY OF REVIEW: The annual risks were 2.1% (CI 95%: 1.9 to 2.4) for nonstroke vascular death, 2.2% (1.7 to 2.7) for total MI, 0.9% (0.7 to 1.2) for nonfatal MI and 1.1% (0.8 to 1.5) for fatal MI. The time course of risk was linear. Estimated risks fitted well with observed risks at the individual level. There was no heterogeneity in the absolute risks according to baseline study characteristics. CONCLUSIONS: Patients with TIA or stroke have a relatively high risk of MI and nonstroke vascular death. Additional research is needed to identify the determinants of coronary artery disease in stroke patients. PMID- 16254219 TI - Mitral annular calcification, aortic valve sclerosis, and incident stroke in adults free of clinical cardiovascular disease: the Strong Heart Study. AB - BACKGROUND AND PURPOSE: Mitral annular calcification (MAC) and aortic valve (AV) sclerosis have each been linked to cardiovascular disease. Whether MAC and AV sclerosis are risk factors for stroke independent of other echocardiographic or laboratory predictors has not been established. We evaluated the relationship between MAC, AV sclerosis, and first stroke events in a population-based cohort. METHODS: Our study cohort consisted of 2723 American Indians participating in the Strong Heart Study who were free of prevalent cardiovascular disease. Participants underwent standardized clinical, echocardiographic, and laboratory evaluation, and incident stroke was ascertained using validated methods. RESULTS: During a median follow-up of 7 years, 86 strokes occurred. Age- and sex-adjusted incidence rates of stroke were significantly increased for MAC (rate ratio [RR], 3.12; 95% CI, 1.77 to 5.25) but not for AV sclerosis (RR, 1.15; 95% CI, 0.45 to 2.49). MAC was also associated with a reduced time to first stroke events after adjustment for clinical variables and the inflammatory markers C-reactive protein and fibrinogen (hazard ratio [HR], 2.42; 95% CI, 1.39 to 4.21) or the echocardiographic covariates left ventricular hypertrophy and left atrial enlargement (HR, 1.89; 95% CI, 1.04 to 3.41). Individuals with and without AV sclerosis showed no significant difference in stroke-free survival in unadjusted analyses (P=0.698). Crossing of the survival curves precluded multivariable analysis using Cox models. CONCLUSIONS: In this cohort of American Indians without clinical cardiovascular disease, the presence of MAC, but not AV sclerosis, proved to be a strong risk factor for incident stroke after extensive adjustment for other predictors. Individuals exhibiting MAC may benefit from aggressive risk factor modification, but this will require further investigation. PMID- 16254220 TI - Characteristics of blood pressure profiles as predictors of long-term outcome after acute ischemic stroke. AB - BACKGROUND AND PURPOSE: Most patients have elevated blood pressure (BP) in the early phase after an acute ischemic stroke. Mechanism and effects of this BP elevation are not well understood. The benefits of intervention by lowering the initial BP or waiting for spontaneous return to normal values remain debated. We studied the hypothesis that increased BP level and profile variability will adversely affect long-term outcome after stroke with and without thrombolytic treatment. METHODS: We studied the 615 patients with acute ischemic hemispheric stroke in the first European Cooperative Acute Stroke Study (ECASS). BP was measured at 2-hour intervals during the first 20 hours after randomization, and then every 4 hours, up to 72 hours after admission. Studied features of individual 0- to 72-hour BP profiles were: baseline BP, maximum and minimum BP, mean level, and successive variation in the BP profile. The end point was good functional recovery (modified Rankin Scale [mRS] score of 0 to 1) at 90 days. Logistic regression was used to adjust for known prognostic factors, demographic, initial stroke severity, disease and medication histories, and computed tomography signs. RESULTS: Higher systolic BP or diastolic BP at baseline were associated with favorable outcome assessed on modified mRS at 90 days (adjusted odds ratio [OR], 1.22; 95% CI, 1.01 to 1.49; and OR, 1.22; 95% CI, 1.01 to 1.49 per 10 mm Hg), lower within-patient 0- to 72-hour average systolic BP (SBP), or DBP implied favorable outcome (OR, 0.74; 95% CI, 0.61 to 0.90; and OR, 0.61; 95% CI, 0.41 to 0.90 per 10 mm Hg). Reduced variability of 0- to 72-hour DBP profile was an independent predictor of favorable outcome (OR, 0.58; 95% CI, 0.39 to 0.85 per 5 mm Hg). CONCLUSIONS: Higher baseline SBP or DBP was associated with favorable outcome after stroke. Other characteristics of first 72-hour BP profiles: lower mean level of SBP or DBP and reduced successive variability of DBP profile were independent predictors of favorable outcome at 90 days. PMID- 16254221 TI - The role of protein kinase C in cerebral ischemic and reperfusion injury. AB - BACKGROUND AND PURPOSE: Stroke is a leading cause of disability and death in the United States, yet limited therapeutic options exist. The need for novel neuroprotective agents has spurred efforts to understand the intracellular signaling pathways that mediate cellular response to stroke. Protein kinase C (PKC) plays a central role in mediating ischemic and reperfusion damage in multiple tissues, including the brain. However, because of conflicting reports, it remains unclear whether PKC is involved in cell survival signaling, or mediates detrimental processes. SUMMARY OF REVIEW: This review will examine the role of PKC activity in stroke. In particular, we will focus on more recent insights into the PKC isozyme-specific responses in neuronal preconditioning and in ischemia and reperfusion-induced stress. CONCLUSIONS: Examination of PKC isozyme activities during stroke demonstrates the clinical promise of PKC isozyme specific modulators for the treatment of cerebral ischemia. PMID- 16254222 TI - Presence of early ischemic changes on computed tomography depends on severity and the duration of hypoperfusion: a single photon emission-computed tomographic study. AB - BACKGROUND AND PURPOSE: To evaluate the clinical significance of early ischemic change (EIC) on computed tomography (CT), pertinent factors that contribute to the appearance of EIC. METHODS: Both CT and technetium-99m hexamethylpropylene amine oxime were performed on a total of 53 patients (34 men, 19 women, mean 69.7 years old) with acute embolic stroke within 6 hours of onset. Patients were excluded if they showed definite clinical recovery or were administered thrombolytic agents. EIC was evaluated using Alberta Stroke Program Early CT Score (ASPECTS). Residual cerebral blood flow (CBF) was determined on SPECT. Variables that were considered pertinent were patients' age, gender, neurological severity, symptom duration, and residual CBF. Using significant pertinent factors for EIC, separate analyses of brain swelling without hypoattenuation and parenchymal hypoattenuation were performed. RESULTS: Patients with EIC (n=37) showed severe neurological deficits, a longer duration and severe hypoperfusion. A positive correlation was observed between ASPECTS and residual CBF (P=0.002; Kruskal-Wallis test). A logistic-regression analysis revealed that both symptom duration (r=0.024, P=0.006) and severity of hypoperfusion (r= -12.167, P=0.006) are independent factors related to EIC. Symptom duration and residual CBF were significantly different among patients with parenchymal hypoattenuation (n=32), brain swelling without hypoattenuation (n=5), and no EIC (P=0.018 and P=0.001, respectively; one-way ANOVA). CONCLUSIONS: The presence of EIC is determined by the duration and the degree of hypoperfusion. This finding supports the hypothesis that tissue damage may be evaluated by a combination of onset time and the presence of EIC. PMID- 16254223 TI - Air pollution and hospital admissions for ischemic and hemorrhagic stroke among medicare beneficiaries. AB - BACKGROUND AND PURPOSE: The association between short-term elevations in ambient air particles and increased cardiovascular morbidity and mortality is well documented. Ambient particles may similarly increase the risk of stroke. METHODS: We evaluated the association between daily levels of respirable particulate matter (aerodynamic diameter < or =10 microm, PM10) and hospital admission for ischemic and hemorrhagic stroke among Medicare recipients (age > or =65 years) in 9 US cities using a 2-stage hierarchical model. In the first stage, we applied the time-stratified case-crossover design to estimate the effect of PM10 in each city. We used a 3-day unconstrained, distributed lag model to simultaneously estimate the effect of PM10 0 to 2 days before the admission day and controlled for meteorological covariates in all of the models. In the second stage, we used random-effects metaanalytic techniques to combine the city-specific effect estimates. RESULTS: Ischemic (n=155,503) and hemorrhagic (19,314) stroke admissions were examined separately. For ischemic stroke, an interquartile range increase in PM10 was associated with a 1.03% (95% CI, 0.04% to 2.04%) increase in admissions on the same day only. Similar results were observed with CO, NO2, and SO2. For hemorrhagic stroke, no association was observed with any pollutant 0 to 2 days before admission. CONCLUSIONS: These results suggest that elevations in ambient particles may transiently increase the risk of ischemic, but not hemorrhagic, stroke. Studies with more accurate assessment of timing of stroke onset are necessary to confirm or refute these findings. PMID- 16254224 TI - Repetitive transcranial magnetic stimulation of contralesional primary motor cortex improves hand function after stroke. AB - BACKGROUND AND PURPOSE: A recent report has demonstrated that the contralesional primary motor cortex (M1) inhibited the ipsilesional M1 via an abnormal transcallosal inhibition (TCI) in stroke patients. We studied whether a decreased excitability of the contralesional M1 induced by 1 Hz repetitive transcranial magnetic stimulation (rTMS) caused an improved motor performance of the affected hand in stroke patients by releasing the TCI. METHODS: We conducted a double blind study of real versus sham rTMS in stroke patients. After patients had well- performed motor training to minimize the possibility of motor training during the motor measurement, they were randomly assigned to receive a subthreshold rTMS at the contralesional M1 (1 Hz, 25 minutes) or sham stimulation. RESULTS: When compared with sham stimulation, rTMS reduced the amplitude of motor-evoked potentials in contralesional M1 and the TCI duration, and rTMS immediately induced an improvement in pinch acceleration of the affected hand, although a plateau in motor performance had been reached by the previous motor training. This improvement in motor function after rTMS was significantly correlated with a reduced TCI duration. CONCLUSIONS: We have demonstrated that a disruption of the TCI by the contralesional M1 virtual lesion caused a paradoxical functional facilitation of the affected hand in stroke patients; this suggests a new neurorehabilitative strategy for stroke patients. PMID- 16254225 TI - The Third Stroke Registry in Tartu, Estonia: decline of stroke incidence and 28 day case-fatality rate since 1991. AB - BACKGROUND AND PURPOSE: The purpose of the present study was to estimate the time trends of stroke during the last 10 years in an Estonian population by comparison of the results from the 2 previous stroke registries from Tartu. METHODS: The Third Stroke Registry in Tartu was conducted from January 12, 2001, to November 30, 2003. The previous registry was composed from January 1, 1991, to December 31, 1993. The design of both studies is similar, using the same study criteria and classification schemes. RESULTS: A total of 1280 patients with first-ever stroke were registered during the 5-year study period. The overall incidence rate of 230 per 100,000 declined between the studies to 188 per 100,000 age standardized to the European standard population. The age-adjusted incidence rate for women decreased from 204 to 164 per 100,000 between the 2 periods. In most of the age groups, the overall case-fatality rates declined during the second period; the trend in the age group 75 to 84 years was statistically significant. CONCLUSIONS: The incidence of first-ever stroke in Tartu has declined significantly during the past decade and reached the level of the first registry. The 28-day case-fatality rate has declined from 30% to 26%. The prevalence of cardiovascular risk factors, incidence of stroke, and ischemic heart disease has been high in Eastern European countries. Our data show that the situation has improved. PMID- 16254226 TI - Association of serum-soluble heat shock protein 60 with carotid atherosclerosis: clinical significance determined in a follow-up study. AB - BACKGROUND AND PURPOSE: Previous work has shown that soluble heat shock protein 60 (HSP60; sHSP60), present in circulating blood, is associated with carotid atherosclerosis. In the current evaluation, we tested the hypothesis that sHSP60 levels are associated with the progression of carotid arteriosclerosis, prospectively. METHODS: The association of sHSP60 with early atherogenesis (5 year development and progression of nonstenotic carotid plaques) was investigated as part of the population-based prospective Bruneck Study. The current study focused on the follow-up period between 1995 and 2000 and, thus, included 684 subjects. RESULTS: sHSP60 levels measured in 1995 and 2000 were highly correlated (r=0.40; P<0.001), indicating consistency over a 5-year period. Circulating HSP60 levels were significantly correlated with antilipopolysaccharide and anti-HSP60 antibodies. It was also elevated in subjects with chronic infection (top quintile group of HSP60, among subjects with and without chronic infection: 23.8% versus 17.0%; P=0.003 after adjustment for age and sex). HSP60 levels were significantly associated with early atherogenesis, both in the entire population (multivariate odds ratio, for a comparison between quintile group V versus I+II: 2.0 [1.2 to 3.5] and the subgroup free of atherosclerosis at the 1995 baseline: 3.8 [1.6 to 8.9]). The risk of early atherogenesis was additionally amplified when high sHSP60 and chronic infection were present together. CONCLUSIONS: Our study provides the first prospective data confirming an association between high levels of sHSP60 and early carotid atherosclerosis. This possibly indicates an involvement of sHSP60 in activating proinflammatory processes associated with early vessel pathology. PMID- 16254227 TI - Evolution of cognitive impairment after stroke and risk factors for delayed progression. AB - BACKGROUND AND PURPOSE: Cognitive decline occurs in approximately 30% of stroke patients. Acute risk factors have been identified, but long-term risk has not been examined in large samples. The purpose of this research was to determine factors associated with the progression of cognitive impairment after stroke. METHODS: Consecutive stroke patients (193) without previous dementia were assessed 3 months after stroke with an extensive neuropsychological battery and diagnosed according to the Diagnostic and Statistical Manual of Mental Disorders Fourth Edition criteria and the Clinical Dementia Rating as normal (139), cognitive decline without dementia (18), or dementia (18 mild, 10 moderate, and 8 severe). After a 24-month follow-up, they were classified as stable, progressing, or improving, according to change in Clinical Dementia Rating score. The determinants of progression of cognitive decline were ascertained by logistic regression analysis of all clinical, neuroimaging, and complementary data. RESULTS: Cognitive status at 24 months was stable in most cases (151; 78.2%), decline progressed in 27 (14%; 6 demented and 21 nondemented), and improved in 15 (7.8%; 7 demented and 8 nondemented). Seven nondemented patients became demented at 24 months, and 5 demented became nondemented. The age (odds ratio [OR], 1.05; 95% CI, 1.01 to 1.1), mental decline before stroke (OR, 1.14; 95% CI, 1.02 to 1.27), number of prescribed drugs (OR, 1.34; 95% CI, 1.05 to 1.72), diastolic blood pressure on admission (OR, 0.96; 95% CI, 0.93 to 0.99), and episodes of hypotension during admission (OR, 7.61; 95% CI, 1.11 to 52.1) were significantly associated with cognitive deterioration. CONCLUSIONS: Cognition is rather stable for 2 years after stroke. Both progression and improvement of cognitive impairment are frequent in demented patients. Age, previous cognitive decline, polypharmacy, and hypotension during admission are risk factors for progression. PMID- 16254228 TI - Dynamic cerebral autoregulation in acute lacunar and middle cerebral artery territory ischemic stroke. AB - BACKGROUND AND PURPOSE: We addressed whether dynamic cerebral autoregulation (dCA) is affected in middle cerebral artery (MCA) territory (MCAS) and lacunar ischemic stroke (LS). METHODS: Blood pressure (MAP) and MCA velocity (V) were measured in 10 patients with large MCAS (National Institutes of Health Stroke score, 17+/-2; mean+/-SEM), in 10 with LS (score, 9+/-1), and in 10 reference subjects. dCA was evaluated in time (delay of the MCA Vmean counter-regulation during changes in MAP) and frequency domains (cross-spectral MCA Vmean-to-MAP phase lead). RESULTS: In reference subjects, latencies for MAP increments (5.3+/ 0.5 seconds) and decrements (5.6+/-0.5 seconds) were comparable, and low frequency MCA Vmean-to-MAP phase lead was 56+/-5 and 59+/-5 degrees (left and right hemisphere). In MCAS, these latencies were 4.6+/-0.7 and 5.6+/-0.5 seconds in the nonischemic hemisphere and not detectable in the ischemic hemisphere. In the unaffected hemisphere, phase lead was 61+/-6 degrees versus 26+/-6 degrees on the ischemic side (P<0.05). In LS, no latency and smaller phase lead bilaterally (32+/-6 and 33+/-5 degrees) conformed to globally impaired dCA. CONCLUSIONS: In large MCAS infarcts, dynamic cerebral autoregulation was impaired in the affected hemisphere. In LS, dynamic cerebral autoregulation was impaired bilaterally, a finding consistent with the hypothesis of bilateral small vessel disease in patients with lacunar infarcts. PMID- 16254229 TI - Improving the health and wellness of persons with disabilities: a call to action. PMID- 16254230 TI - State and metropolitan-area estimates of disability in the United States, 2001. AB - OBJECTIVES: We sought to provide estimates of disability prevalence for states and metropolitan areas in the United States. METHODS: We analyzed Behavioral Risk Factor Surveillance System data from 2001 for all 50 states and the District of Columbia and 103 metropolitan areas. We performed stratified analyses by demographics for 20 metropolitan areas with the highest prevalence of disability. RESULTS: State disability estimates ranged from 10.5% in Hawaii to 25.9% in Arizona. Metropolitan disability estimates ranged from 10.2% in Honolulu, Hawaii to 27.1% in Tucson, Ariz. Regional metropolitan medians for disability (range, 17.0-19.7%) were similar across the Northeast, Midwest, and South and were highest in the West. In the 20 metropolitan areas with the highest disability estimates, the prevalence of disability generally increased with age and was higher for women and those with a high-school education or less. CONCLUSIONS: State and metropolitan-area estimates may be used to guide state and local efforts to prevent, delay, or reduce disability and secondary conditions in persons with disabilities. PMID- 16254231 TI - Widening social inequalities in risk for sudden infant death syndrome. AB - OBJECTIVES: In 1994, the US Public Health Service launched the "Back to Sleep" campaign, promoting the supine sleep position to prevent sudden infant death syndrome (SIDS). Studies of SIDS in the United States have generally found socioeconomic and race disparities. Our objective was to see whether the "Back to Sleep" campaign, which involves an effective, easy, and free intervention, has reduced social class inequalities in SIDS. METHODS: We conducted a population based case-cohort study during 2 periods, 1989 to 1991 and 1996 to 1998, using the US Linked Birth/Infant Death Data Sets. Case group was infants who died of SIDS in infancy (N = 21 126); control group was a 10% random sample of infants who lived through the first year and all infants who died of other causes (N=2241218). Social class was measured by mother's education level. RESULTS: There was no evidence that inequalities in SIDS were reduced after the Back to Sleep campaign. In fact, odds ratios for SIDS associated with lower social class increased between 1989-1991 and 1996-1998. The race disparity in SIDS increased after the Back to Sleep campaign. CONCLUSIONS: The introduction of an inexpensive, easy, public health intervention has not reduced social inequalities in SIDS; in fact, the gap has widened. Although the risk of SIDS has been reduced for all social class groups, women who are more educated have experienced the greatest decline. PMID- 16254232 TI - Incidence of disability among preretirement adults: the impact of depression. AB - OBJECTIVES: We evaluated the effect of depression on risk, on the basis of standardized assessment, for developing activities of daily living (ADL) disability. METHODS: Depression-related risk on 2-year ADL disability is estimated from 6871 participants in a population-based national sample aged 54-65 years and free of baseline ADL disability. We evaluated the effects of factors amenable to clinical and public health intervention that may explain the relationship between depression and incident disability. RESULTS: The odds of ADL disability were 4.3 times greater for depressed adults than their non-depressed peers (95% confidence interval=3.1, 6.0). Among depressed adults, 18.7% of African Americans, 8.0% of Whites, and 7.8% of His-panics developed disability within 2 years. The attributable population fraction because of depression is 17.3% (95% confidence interval=11%, 24%). Concurrent health factors moderated depression-associated risk. CONCLUSIONS: Elevated risk of ADL disability onset because of depression, in a cohort whose medical costs will imminently be covered via Medicare, is attenuated by factors amenable to public health intervention. Prevention and/or public health/policy programs that lead to more accessible and effective mental health and medical care could reduce the development of ADL disability among depressed adults. PMID- 16254233 TI - Mental disorder and violent victimization in a total birth cohort. AB - OBJECTIVE: We examined the association between mental disorder and violent victimization in a general population sample. METHODS: We performed a multivariate analysis of violent victimization in a 12-month period on a total birth cohort with follow-up data that assessed, during their 21st year, males and females born in Dunedin, New Zealand, in the early 1970s. RESULTS: Compared with people with no mental disorder, (1) people with anxiety disorders experienced more sexual assaults, (2) people with schizophreniform disorders experienced more threatened and completed physical assaults, (3) people with alcohol dependence disorders experienced more completed physical assaults, and (4) people with marijuana dependence disorders experienced more attempted physical assaults. These results held after control for psychiatric comorbidity, demographic characteristics, and the study participants' own violent behavior. CONCLUSION: Mentally disordered young adults tend to experience more violent victimization in the community than those without a mental disorder. PMID- 16254234 TI - Accessibility of health clubs for people with mobility disabilities and visual impairments. AB - OBJECTIVE: We sought to examine the accessibility of health clubs to persons with mobility disabilities and visual impairments. METHODS: We assessed 35 health clubs and fitness facilities as part of a national field trial of a new instrument, Accessibility Instruments Measuring Fitness and Recreation Environments (AIMFREE), designed to assess accessibility of fitness facilities in the following domains: (1) built environment, (2) equipment, (3) swimming pools, (4) information, (5) facility policies, and (6) professional behavior. RESULTS: All facilities had a low to moderate level of accessibility. Some of the deficiencies concerned specific Americans with Disabilities Act guidelines pertaining to the built environment, whereas other deficiency areas were related to aspects of the facilities' equipment, information, policies, and professional staff. CONCLUSIONS: Persons with mobility disabilities and visual impairments have difficulty accessing various areas of fitness facilities and health clubs. AIMFREE is an important tool for increasing awareness of these accessibility barriers for people with disabilities. PMID- 16254237 TI - Cell migration at a glance. PMID- 16254238 TI - Talin phosphorylation sites mapped by mass spectrometry. PMID- 16254235 TI - Persistent and growing socioeconomic disparities in disability among the elderly: 1982-2002. AB - OBJECTIVE: We sought to determine whether socioeconomic and racial/ethnic disparities in prevalence of disability over age 70 have widened or narrowed during the past 2 decades. METHODS: We used data from the 1982-2002 National Health Interview Surveys, which are nationally representative cross-sectional surveys of the noninstitutionalized population of the United States. Participants included 172227 people aged 70 years and older. The primary outcome measure was the average annual percentage change in the prevalence of 2 self-reported disability measures: the need for help with activities of daily living ("ADL disability") and need for help with either ADL or instrumental activities of daily living ("any disability"). RESULTS: All groups experienced declines in the age- and gender-adjusted prevalence of any disability during the 1982 to 2002 period. However, the average annual percent declines were smaller for the least advantaged socioeconomic groups. Differences in trends across racial/ethnic groups were not statistically significant. ADL disability prevalence decreased for the more advantaged groups but increased among the lowest income and education groups. Non-Hispanic Whites and minorities experienced similar average annual percent declines in ADL disability. CONCLUSIONS: Racial/ethnic disparities in old-age disability have persisted over the last 20 years, whereas socioeconomic disparities have increased. PMID- 16254239 TI - Paxillin phosphorylation sites mapped by mass spectrometry. PMID- 16254240 TI - FAK phosphorylation sites mapped by mass spectrometry. PMID- 16254241 TI - CZH proteins: a new family of Rho-GEFs. AB - The Rho family of small GTPases are important regulators of multiple cellular activities and, most notably, reorganization of the actin cytoskeleton. Dbl homology (DH)-domain-containing proteins are the classical guanine nucleotide exchange factors (GEFs) responsible for activation of Rho GTPases. However, members of a newly discovered family can also act as Rho-GEFs. These CZH proteins include: CDM (Ced-5, Dock180 and Myoblast city) proteins, which activate Rac; and zizimin proteins, which activate Cdc42. The family contains 11 mammalian proteins and has members in many other eukaryotes. The GEF activity is carried out by a novel, DH-unrelated domain named the DOCKER, CZH2 or DHR2 domain. CZH proteins have been implicated in cell migration, phagocytosis of apoptotic cells, T-cell activation and neurite outgrowth, and probably arose relatively early in eukaryotic evolution. PMID- 16254242 TI - Scale-free networks in cell biology. AB - A cell's behavior is a consequence of the complex interactions between its numerous constituents, such as DNA, RNA, proteins and small molecules. Cells use signaling pathways and regulatory mechanisms to coordinate multiple processes, allowing them to respond to and adapt to an ever-changing environment. The large number of components, the degree of interconnectivity and the complex control of cellular networks are becoming evident in the integrated genomic and proteomic analyses that are emerging. It is increasingly recognized that the understanding of properties that arise from whole-cell function require integrated, theoretical descriptions of the relationships between different cellular components. Recent theoretical advances allow us to describe cellular network structure with graph concepts and have revealed organizational features shared with numerous non biological networks. We now have the opportunity to describe quantitatively a network of hundreds or thousands of interacting components. Moreover, the observed topologies of cellular networks give us clues about their evolution and how their organization influences their function and dynamic responses. PMID- 16254243 TI - Set1- and Clb5-deficiencies disclose the differential regulation of centromere and telomere dynamics in Saccharomyces cerevisiae meiosis. AB - The entry into meiosis is characterized by a lengthy premeiotic S phase and a reorganization of the nuclear architecture. Analysis of centromere and telomere dynamics in wild-type Saccharomyces cerevisiae meiosis suggests that resolution of vegetative centromere and telomere clusters are independent events differently connected to premeiotic S phase. Absence of the B-type cyclin Clb5 or the Set1 histone methyltransferase leads to a delay of premeiotic S phase by separate mechanisms. In clb5Delta cells, centromere cluster resolution appears normal, whereas dissolution of the vegetative telomere clusters is impaired and meiosis specific clustering of telomeres, i.e. bouquet formation, is grossly delayed. In set1Delta cells, centromere and telomere redistribution are both impaired and bouquet nuclei are absent, despite proper location of the meiosis-specific telomere protein Ndj1. Thus, centromere and telomere redistribution at the onset of prophase I is differentially regulated, with centromere dispersion occurring independently of premeiotic S phase. The normal kinetics of dissolution of the vegetative telomere clusters in a set1Delta mec1-1 mutant suggests the presence of a checkpoint that limits the dispersion of telomeres in absence of Set1. PMID- 16254244 TI - Nuclear levels and patterns of histone H3 modification and HP1 proteins after inhibition of histone deacetylases. AB - The effects of the histone deacetylase inhibitors (HDACi) trichostatin A (TSA) and sodium butyrate (NaBt) were studied in A549, HT29 and FHC human cell lines. Global histone hyperacetylation, leading to decondensation of interphase chromatin, was characterized by an increase in H3(K9) and H3(K4) dimethylation and H3(K9) acetylation. The levels of all isoforms of heterochromatin protein, HP1, were reduced after HDAC inhibition. The observed changes in the protein levels were accompanied by changes in their interphase patterns. In control cells, H3(K9) acetylation and H3(K4) dimethylation were substantially reduced to a thin layer at the nuclear periphery, whereas TSA and NaBt caused the peripheral regions to become intensely acetylated at H3(K9) and dimethylated at H3(K4). The dispersed pattern of H3(K9) dimethylation was stable even at the nuclear periphery of HDACi-treated cells. After TSA and NaBt treatment, the HP1 proteins were repositioned more internally in the nucleus, being closely associated with interchromatin compartments, while centromeric heterochromatin was relocated closer to the nuclear periphery. These findings strongly suggest dissociation of HP1 proteins from peripherally located centromeres in a hyperacetylated and H3(K4) dimethylated environment. We conclude that inhibition of histone deacetylases caused dynamic reorganization of chromatin in parallel with changes in its epigenetic modifications. PMID- 16254245 TI - Differential contributions of connexin37 and connexin43 to oogenesis revealed in chimeric reaggregated mouse ovaries. AB - The gap junction proteins connexin37 and connexin43 are required for ovarian folliculogenesis in the mouse. To define their respective roles in oogenesis, chimeric ovaries containing either null mutant oocytes and wild-type granulosa cells or the reverse combination were grafted to the renal capsules of immunodeficient female mice. After three weeks, the oocytes were tested for meiotic competence and fertilizability in vitro. Ovaries composed of connexin43 deficient oocytes and wild-type granulosa cells produced antral follicles enclosing oocytes that could develop to at least the two-cell stage, demonstrating that oocytes need not express connexin43 to reach maturity. Conversely, both follicle development and oocyte maturation were impaired in ovaries containing either wild-type oocytes and connexin43-deficient granulosa cells or connexin37-deficient oocytes and wild-type granulosa cells. Thus absence of connexin43 from granulosa cells or connexin37 from oocytes is sufficient to compromise both oocyte and follicle development. Wild-type oocytes paired with connexin37-deficient granulosa cells generated antral follicles containing oocytes that developed to at least the two-cell stage. Therefore, connexin37 absence from granulosa cells need not impair fertility in mice. Dye transfer experiments revealed persistent oocyte-granulosa cell coupling in those follicles, indicating functional compensation by another connexin. The results indicate that mouse oocytes do not need to express connexin43 in order to develop into meiotically competent, fertilizable gametes, but must express connexin37 for communication with granulosa cells, a requirement for oogenesis. PMID- 16254246 TI - The JIL-1 kinase interacts with lamin Dm0 and regulates nuclear lamina morphology of Drosophila nurse cells. AB - We have used a yeast two-hybrid screen to identify lamin Dm0 as an interaction partner for the nuclear JIL-1 kinase. This molecular interaction was confirmed by GST-fusion protein pull-down assays and by co-immunoprecipitation experiments. Using deletion construct analysis we show that a predicted globular domain of the basic region of the COOH-terminal domain of JIL-1 was sufficient for mediating the molecular interactions with lamin Dm0. A reciprocal analysis with truncated lamin Dm0 constructs showed that the interaction with JIL-1 required sequences in the tail domain of lamin Dm0 that include the Ig-like fold. Further support for a molecular interaction between JIL-1 and lamin Dm0 in vivo was provided by genetic interaction assays. We show that nuclear positioning and lamina morphology were abnormal in JIL-1 mutant egg chambers. The most common phenotypes observed were abnormal nurse cell nuclear lamina protrusions through the ring canals near the oocyte, as well as dispersed and mislocalized lamin throughout the egg chamber. These phenotypes were completely rescued by a full-length JIL-1 transgenic construct. Thus, our results suggest that the JIL-1 kinase is required to maintain nuclear morphology and integrity of nurse cells during oogenesis and that this function may be linked to molecular interactions with lamin Dm0. PMID- 16254247 TI - Phosphorylation of Nlp by Plk1 negatively regulates its dynein-dynactin-dependent targeting to the centrosome. AB - When cells enter mitosis the microtubule (MT) network undergoes a profound rearrangement, in part due to alterations in the MT nucleating and anchoring properties of the centrosome. Ninein and the ninein-like protein (Nlp) are centrosomal proteins involved in MT organisation in interphase cells. We show that the overexpression of these two proteins induces the fragmentation of the Golgi, and causes lysosomes to disperse toward the cell periphery. The ability of Nlp and ninein to perturb the cytoplasmic distribution of these organelles depends on their ability to interact with the dynein-dynactin motor complex. Our data also indicate that dynactin is required for the targeting of Nlp and ninein to the centrosome. Furthermore, phosphorylation of Nlp by the polo-like kinase 1 (Plk1) negatively regulates its association with dynactin. These findings uncover a mechanism through which Plk1 helps to coordinate changes in MT organisation with cell cycle progression, by controlling the dynein-dynactin-dependent transport of centrosomal proteins. PMID- 16254248 TI - Dimerization of the cytokine receptors gp130 and LIFR analysed in single cells. AB - The cytokine receptor gp130 is the shared signalling subunit of the IL-6-type cytokines. Interleukin-6 (IL-6) signals through gp130 homodimers whereas leukaemia inhibitory factor (LIF) exerts its action through a heterodimer of gp130 and the LIF receptor (LIFR). Related haematopoietic receptors such as the erythropoietin receptor have been described as preformed dimers in the plasma membrane. Here we investigated gp130 homodimerization and heterodimerization with the LIFR by fluorescence resonance energy transfer (FRET) and bimolecular fluorescence complementation (BiFC). We detected a FRET signal between YFP- and CFP-tagged gp130 at the plasma membrane of unstimulated cells that does not increase upon IL-6 stimulation. However, FRET between YFP-tagged gp130 and CFP tagged LIFR considerably increased upon LIF stimulation. Using a BiFC approach that detects stable interactions we show that fluorescence complementation of gp130 constructs tagged with matching 'halves' of fluorescent proteins increases upon IL-6 stimulation. Taken together, these findings suggest that transient gp130 homodimers on the plasma membrane are stabilized by IL-6 whereas heterodimerization of gp130 with the LIFR is mainly triggered by the ligand. This view is supported by the observation that the simultaneous action of two IL-6 binding domains on two gp130 molecules is required to efficiently recruit a fluorescent IL-6 (YFP-IL-6) to the plasma membrane. PMID- 16254249 TI - Assigning functions to distinct regions of the N-terminus of the prion protein that are involved in its copper-stimulated, clathrin-dependent endocytosis. AB - The cellular prion protein (PrP(C)) is essential for the pathogenesis and transmission of prion diseases. Although PrP(C) is known to be located in detergent-insoluble lipid rafts at the surface of neuronal cells, the mechanism of its internalisation is unclear, with both raft/caveolae-based and clathrin mediated processes being proposed. We have investigated the mechanism of copper induced internalisation of PrP(C) in neuronal cells by immunofluorescence microscopy, surface biotinylation assays and buoyant sucrose density gradient centrifugation in the presence of Triton X-100. Clathrin-mediated endocytosis was selectively blocked with tyrphostin A23, which disrupts the interaction between tyrosine motifs in the cytosolic domains of integral membrane proteins and the adaptor complex AP2, and a dominant-negative mutant of the adaptor protein AP180. Both these agents inhibited the copper-induced endocytosis of PrP(C). Copper caused PrP(C) to move laterally out of detergent-insoluble lipid rafts into detergent-soluble regions of the plasma membrane. Using mutants of PrP(C) that lack either the octapeptide repeats or the N-terminal polybasic region, and a construct with a transmembrane anchor, we show that copper binding to the octapeptide repeats promotes dissociation of PrP(C) from lipid rafts, whereas the N-terminal polybasic region mediates its interaction with a transmembrane adaptor protein that engages the clathrin endocytic machinery. Our results provide an experimental basis for reconciling the apparently contradictory observations that the prion protein undergoes clathrin-dependent endocytosis despite being localised in lipid rafts. In addition, we have been able to assign distinct functions in the endocytic process to separate regions of the protein. PMID- 16254251 TI - Cigarette smoking induces overexpression of hepatocyte growth factor in type II pneumocytes and lung cancer cells. AB - We examined gene expression of hepatocyte growth factor (HGF) and HGF receptor (HGFR), or product of proto-oncogene c-met (c-met), in smokers and nonsmokers with adenocarcinoma (ADC) by suppression subtractive hybridization and microarray techniques. Expression of HGF and c-met was confirmed by RT-PCR. HGF content in the respective tumor mass and nontumor lung tissue was measured by ELISA. HGF in pathologic samples was localized by immunohistochemistry and in situ hybridization. Our results indicate that overexpression of HGFR was frequently detected in ADC cells, whereas overexpression of HGF was detected in alveolar type II (ATII) cells. Overexpression of HGF was correlated with cigarette smoking and tumor stages. In vitro, HGF expression was evaluated in isolated murine ATII cells and in 12 ADC cell lines, and we found that nicotine activated HGF expression in ATII cells and lung cancer cells. PMID- 16254252 TI - Cardiovascular complications of non-steroidal anti-inflammatory drugs. AB - Coxibs, such as rofecoxib, celecoxib, and valdecoxib, selectively inhibit cyclooxygenase (COX)-2, the mainly inducible, pro-inflammatory COX isoform. Unlike traditional non-steroidal anti-inflammatory drugs (NSAIDs) most coxibs do not significantly inhibit COX-1 and are therefore less toxic to the gastrointestinal tract. Hence, coxibs widely replaced traditional NSAIDs for treatment of arthritis and other painful inflammatory conditions. In many, but not all, clinical studies, coxibs became associated with higher risks of myocardial infarction (MI) and stroke. Several mechanisms may be involved in the pathogenesis of such complications. First, selective inhibition of COX-1 lowers platelet synthesis of thromboxane (TXA(2)), a thrombogenic and atherogenic eicosanoid. Selective inhibition of COX-2 limits endothelial cell synthesis of prostacyclin (PGI(2)), an arachidonic acid product that opposes the effects of thromboxane. In apoE-/- mice, interruption of TXA(2) signaling by deletion of its receptor (TP) limits atherogenesis, whereas interruption of PGI2 signaling by deletion of its receptor (IP) accelerates atherogenesis. This suggests that selective inhibition of COX-2 can disrupt the physiological balance between thromboxane and prostacyclin and thus increase atherosclerosis, thrombogenesis, and the risk of cardiovascular complications. Second, COX inhibition can raise levels of arachidonic acid, which can inhibit mitochondrial oxidative phosphorylation (OXPHOS) and increase OXPHOS generation of reactive oxygen species. Several NSAIDs, including coxibs and meloxicam, directly uncouple or inhibit OXPHOS. Studies of apoE-/- mice indicate that mitochondrial dysfunction plays an early role in atherogenesis. Third, many NSAIDs exhibit COX-independent properties. For example, in animal models, short-term treatment with celecoxib reduces monocyte chemotaxis by reducing expression of monocyte chemoattractant protein (MCP)-1. However, long-term treatment results in the opposite effect and accelerates atherogenesis. In conclusion, to reduce the risk of cardiovascular complications during long-term coxib therapy, low-dose aspirin supplementation should be considered. An alternative is to use a less COX-2-selective inhibitor such as meloxicam. Genotyping of -765 alleles of the COX-2 gene promoter and examining the polymorphism of other genes involved in eicosanoid metabolism or NSAID degradation may become helpful in predicting patients who are at higher risk of cardiovascular complications during selective COX-2 inhibitor therapy. PMID- 16254253 TI - The autopsy in a tertiary teaching hospital in Brazil. AB - A high autopsy rate allows accurate epidemiological studies and quality control of medical care. This study aims to analyze all autopsies performed in a university teaching hospital in Brazil during 52 consecutive wk. The following data were retrieved from individual autopsy records: gender, age, time of death (hr, day, and month), and the main cause of death. There were 1419 autopsies (79% adults and 21% pediatrics, 60% male and 40% female). Those performed during working days summed up to 67.5%, the remaining 32.5% were performed during weekends or holidays. Autopsies were more frequent during the nocturnal than diurnal period (52.6% vs 47.4%, respectively). The causes of death distributed among the ICD-10 categories were: cardiovascular diseases 21.3%, infectious diseases 19.2%, neoplasms 12.8%, perinatal conditions 10.8%, respiratory diseases 6.6%, gastrointestinal diseases 6.0%, congenital anomalies 4.7%, CNS diseases 3.8%, genitourinary diseases 1.8%, and others 13.0%. There was coexistence of the diseases typical of both industrialized and developing countries, indicating the epidemiological transition in our country. Our data indicate that staff supervision of undergraduate medical students and residents is adequate. Along with a brief discussion of the historical, cultural, and legal factors that allow a high autopsy rate, this study reinforces the importance of the autopsy in a tertiary teaching hospital. PMID- 16254254 TI - Apolipoprotein E genotype in matched men and women with coronary heart disease. AB - Apolipoprotein E (apo E) plays an important role in lipid metabolism and its polymorphism may be a risk determinant of coronary heart disease (CHD). Since evidence suggested a gender-specific effect of apo E polymorphism, we studied the influence of gender-specific interaction of the polymorphism on CHD. From a total of 463 Greek Caucasians (314 men and 149 postmenopausal women) with angiographically documented CHD, we selected 79 women (68+/- 9 yr old) and 79 men (66+/- 9 yr old) who were matched for clinical characteristics. Apo E genotyping was performed by PCR and RFLP analysis. Biochemical parameters were also measured. The results were as follows: the E3/3 genotype occurred in 78.5% of the patients, followed by E3/4, E2/3, E2/4, and E4/4 genotypes, which occurred in 9.5%, 9.5%, 1.9%, and 0.6% of the patients, respectively. No significant differences were observed in the apo E allele or apo E genotype distributions between the matched Greek men and women with CHD. The E3/3 men patients were more frequently part of a family with a history of CHD, compared to women (p=0.035). PMID- 16254255 TI - Histone deacetylase inhibitor pharmacodynamic analysis by multiparameter flow cytometry. AB - Histone deacetylase (HDAC) inhibitors are a promising new class of anticancer drug. The aim of this study was to develop a versatile and sensitive technique for the pharmacodynamic (PD) assessment of HDAC inhibitor activity as monotherapy and in combination therapy. A multiparameter flow cytometric assay was developed initially in healthy donor lymphocytes and leukemia cell lines, and then tested in peripheral blood of solid tumor patients and in bone marrow aspirates of leukemia patients on phase I trials of the HDAC inhibitor MS-275. A technique was developed that allows highly sensitive single parameter determination of HDAC inhibitor activity in as little as 50 microl of whole blood. Multiparameter analysis enabled correlation on a single cell basis of protein acetylation with biologically relevant markers including cell lineage antigens, an apoptosis marker, and PD markers of other anti-cancer agents. The level of protein acetylation can be readily detected and quantified in peripheral blood or in bone marrow aspirates by flow cytometric analysis. The technique described has significant advantages for the PD assessment of HDAC inhibitors as monotherapy and as a component of combination therapy trials. PMID- 16254256 TI - Significance of cerebrospinal fluid sIL-2R level as a marker of CNS involvement in acute lymphoblastic leukemia. AB - Soluble IL-2 receptor (sIL-2R), total protein, uric acid, glucose, aspartate aminotransferase (AST) and lactate dehydrogenase (LD) levels were analyzed in 153 (19 cytology(+), 134 cytology(-)) pairs of CSF and serum samples and the data were compared with the results of cytologic examination to find new CSF markers of CNS involvement in 77 patients with acute lymphoblastic leukemia (ALL). The CSF leukocyte count of cytology(+) samples averaged 107.6+/-362.4 cells/microl, and was higher than that of cytology(-) samples (1.0+/-3.4 cells/microl, p=0.001). The CSF sIL2-R level of cytology(+) samples averaged 162.1+/-247.7 U/ml, and was higher than that of cytology(-) samples (11.2+/-44.6 U/ml, p <0.001). The CSF total protein, uric acid, glucose, AST, and LD levels were not significantly different in cytology(+) and cytology(-) samples (p >0.05). ROC curves showed that the discrimination power of CSF sIL2-R for the presence of leukemic blasts was better than that of CSF leukocyte counts. With a cut-off value for CSF sIL2-R at 10 U/ml, the sensitivity was 89.5% and the specificity was 89.6%. With a cut-off value for CSF leukocyte count at 4 cells/microl, the sensitivity and specificity were 47.4% and 63.2%, respectively. In conclusion, CSF sIL2-R level is a valuable marker of CNS involvement in ALL patients; a level of >10 U/ml may serve as an objective indicator of CNS involvement in conjunction with conventional cytology and the CSF leukocyte count. PMID- 16254257 TI - p-p70S6K (Thr 389) Expression in nodular sclerosing hodgkin's disease as evidence for receptor tyrosine kinase signaling. AB - Platelet-derived growth factor receptor-alpha (PDGFR-alpha) expression in the Hodgkin/Reed-Sternberg cells of nodular sclerosing Hodgkin's disease has been demonstrated in 2 studies. Additional receptor tyrosine kinases have also now been documented. This communication reports the correlative expression in nodular sclerosing Hodgkin's disease of activated (phosphorylated) p70S6K, one of the putative downstream effectors common to receptor tyrosine kinase signaling. PMID- 16254258 TI - Association between plasma interleukin-18 levels and liver injury in chronic hepatitis C virus infection and non-alcoholic fatty liver disease. AB - There is significant upregulation of interleukin-18 (IL-18) expression in viral infectious diseases and in some chronic hepatic diseases, especially (i) hepatitis C virus (HCV) infection, (ii) HCV infection with persistently normal ALT levels (PNAL), and (iii) non-alcoholic fatty liver disease (NAFLD). The aim of this study was a better understanding of the implications of plasma IL-18 levels in the above-mentioned liver diseases. Thirty-four patients with HCV infection, 13 with NAFLD, and 10 controls were enrolled. The HCV-RNA and HCV genotypes and the serum or plasma levels of IL-18, aspartate aminotransferase (AST), alanine aminotransferase (ALT), gamma-glutamyltranspeptidase (gamma-GT), alkaline phosphatase, total cholesterol, triglycerides, alpha(1)-fetoprotein, and ferritin were evaluated. Patients with HCV showed higher levels of IL-18 than the NAFLD patients (p <0.01) and the controls (p <0.005). Patients with NAFLD showed higher values of body mass index and liver disease parameters, compared to HCV infected subjects or controls. These data confirm previous reports of enhanced expression of IL-18 in patients with HCV and NAFLD, compared to healthy subjects, and suggest that IL-18 is important as a marker of liver diseases. PMID- 16254259 TI - Serum magnesium levels and acute exacerbation of chronic obstructive pulmonary disease: a retrospective study. AB - A decrease in serum Mg(+2) is associated with airway hyper-reactivity and impaired pulmonary function. The purpose of this study was to determine if decreased serum Mg(+2) levels in patients with chronic obstructive pulmonary disease (COPD) are associated with acute exacerbations. In a retrospective study, the charted serum Mg(+2) levels in 100 COPD patients were examined. These included 50 patients who presented with an acute exacerbation of COPD and 50 stable patients. Chart review was sequential within both groups. Serum Mg(2+) levels in the stable COPD patients averaged 0.91+/- 0.10 mmol/L (mean+/- SD) with a 95% CI of 0.88 - 0.94 mmol/L. Patients undergoing an exacerbation had significantly lower serum Mg(+2) levels (0.77+/- 0.10 mmol/L; CI, 0.74 - 0.79; p<0.0001). Logistic regression of the dichotomous outcomes as a function of serum Mg(+2) concentration demonstrated a highly significant association (chi(2)=41.26; p<10(-5)). These data were subjected to receiver-operator characteristic (ROC) analysis for decision levels (DL) and the area under the ROC curve was determined to be 0.85+/- 0.04 (CI, 0.78 - 0.93). The optimum DL was determined to lie between 0.80 mmol/L (OR=14.33; sensitivity 70%; specificity 86%) and 0.84 mmol/L (OR=11.16; sensitivity 84%; specificity 68%). These data suggest that at the lower range of the reference interval, serum Mg(+2) levels are associated with an increased risk of exacerbation of symptoms in COPD patients. Furthermore, they suggest a DL that is useful for predicting clinical outcomes in these patients and serving as a target value for therapy. PMID- 16254260 TI - Relationships of lead, copper, zinc, and cadmium levels versus hematopoiesis and iron parameters in healthy adolescents. AB - To investigate the relationships of trace element concentrations vs hematopoiesis and iron parameters, we measured lead, copper, zinc, cadmium, and ferritin levels in 251 healthy adolescents. Concentrations of trace metals were determined by atomic absorption spectrophotometry. There were no significant gender-related differences in serum copper or serum cadmium concentrations. However, blood lead and serum zinc levels were significantly higher in males than females (3.82+/- 1.24 and 118.4 +/- 43.7 microg/dl vs 2.86+/- 1.06 and 83.5+/- 35.2 microg/dl, p<0.05, respectively). Subjects with elevated lead and copper concentrations exhibited significantly higher leukocyte counts and significantly lower serum iron levels than those with decreased lead and copper concentrations, but no significant differences were observed in blood erythrocyte counts or hemoglobin levels between the 2 groups. Blood lead concentrations were 2-fold higher in male adolescents with leukocytes >9.1 x 10(3)/microl than in those with leukocytes <4.3 x 10(3)/microl (5.04+/- 1.67 microg/dl vs 2.51+/- 0.75 microg/dl, p<0.05). Leukocyte counts had significant correlations with blood lead (r=0.39, p<0.05) and serum copper (r=0.26, p<0.05) in males and zinc (r=0.28, p <0.05) in females. Serum iron levels were inversely correlated with blood lead and serum copper concentrations but were not correlated with serum zinc or cadmium levels. In short, blood lead and serum copper concentrations have important relationships to leukocyte counts and iron parameters in adolescents. PMID- 16254261 TI - Sensitivity, specificity, and predictive value of serum soluble transferrin receptor at different stages of iron deficiency. AB - This study investigated the efficiency of serum soluble transferrin receptor (sTfR) for assessing body iron status at different stages of iron deficiency. Among 72 patients with advanced iron-deficiency anemia (IDA), the sensitivity and specificity of sTfR (at a diagnostic cutoff of 3.24 mg/L) were 70.8% and 90.6%, respectively, with a positive predictive value of 85.0%. Sensitivities of sTfR in patients at the earliest stage of iron deficiency (n=41) and the intermediate stage of iron-deficient erythropoiesis (n=15) were 21.9% and 26.7%, respectively, at the same cutoff value of sTfR. Serum ferritin concentrations averaged 6.7+/ 1.9 microg/L in IDA patients with sTfR <3.24 mg/L, which were significantly above the values in IDA patients with sTfR >or=3.24 mg/L (4.8+/-1.2 microg/L, p<0.05). In healthy controls, blood reticulocyte counts were significantly higher in subjects with sTfR >or=3.24 mg/L than in those with sTfR <3.24 mg/L (0.045+/- 0.013 (10(12)/L) vs 0.034+/- 0.011 (10(12)/L), p<0.05]. In conclusion, sTfR level is not a sensitive indicator for the early or intermediate stages of iron deficiency, although sTfR assay can be a useful aid in the diagnosis of advanced IDA. Serum sTfR concentration has significant relationships with blood reticulocyte counts in healthy subjects and with serum ferritin levels in IDA patients. PMID- 16254262 TI - In vivo effects of caffeic acid phenethyl ester on myocardial ischemia reperfusion injury and apoptotic changes in rats. AB - Ischemia/reperfusion (I/R) has been reported to induce apoptotic cellular death in myocardium. This study tested the hypothesis that caffeic acid phenethyl ester (CAPE), one of the active components of propolis, may ameliorate myocardial apoptosis and oxidative myocardial injury. Wistar rats were divided into 4 groups: (i) sham operated, (ii) I/R, (iii) I/R+CAPE, and (iv) I/R+glutathione (GSH). CAPE (10 micromol/kg) was infused iv 10 min before occlusion of the left anterior descending coronary artery (30 min) followed by reperfusion (120 min). GSH (5 mg/kg) was infused iv after the occlusion and immediately before reperfusion. The TdT-mediated in situ nick end-labeling (TUNEL) method was used to evaluate apoptotic activity. I/R resulted in myocardial apoptosis, alterations of antioxidant status, elevation of serum creatine kinase (CK) and aspartate aminotransferase (AST) activities, evidence of lipid peroxidation, and elevated nitric oxide levels, compared to the sham-operation group. No apoptotic cells were found in the myocardial tissue of sham-operated rats. The TUNEL-positive myocardial cells averaged 60%, 30%, and 40% in the I/R, I/R+CAPE, and I/R+GSH groups, respectively. This study demonstrates that pretreatment with CAPE provides cardio-protection from I/R injury. The I/R+CAPE group showed reduced apoptosis, attenuated NO production, elevated myocardial superoxide dismutase (SOD) activity, and diminished serum CK and AST activities, compared to the I/R group. PMID- 16254263 TI - Effect of femoral nutrient artery ligature on radionuclide uptake in rabbits. AB - To elucidate the effect of damaging the nutrient artery to a long bone, we used radionuclide imaging to survey the change of femoral blood supply after ligating the nutrient artery in rabbits. The radionuclide uptake of the femoral shaft, upper and lower metaphysis, and epiphysis decreased significantly at 1 hr postoperation (0.66, 0.74, 0.81, respectively, p <0.05), were close to normal on day 4 (0.96, 0.98, 1.02, respectively, p >0.05), slightly exceeded the contralateral level at 9 and 12 days, and returned to normal on day 16. This study shows that ligation of the nutrient artery of a long bone leads to an immediate decrease in the bone blood flow. However, so long as anastomoses among other bone vessels are intact, the recovery rate is speedy, and long-term, serious disorders of the bone blood supply do not ensue. PMID- 16254264 TI - Comparison of cardiac troponin I levels in serum and sodium-citrate plasma using the ACCESS 2 immunoassay. AB - The aim of this study was to determine if sodium-citrate plasma is a suitable substitute for serum for cardiac troponin I (cTnI) measurement using the Beckman ACCESS 2 immunoassay. Linear regression analysis of cTnI levels in paired serum and sodium-citrate plasma samples from 80 patients provided a y-intercept of 0.0193 and a slope of 1.044. The correlation was good [r=0.995; 95% confidence interval 1.017 to 1.068, p <0.0001]. The paired results were not significantly different by Student's t-test. Serum and sodium-citrate plasma appear to be interchangeable as samples for cTnI measurement using the ACCESS 2 assay. Furthermore, the use of plasma reduces the turnaround time and may avoid false positive cTnI results due to residual fibrin. PMID- 16254265 TI - Preservation of clinical isolates of Mycobacterium tuberculosis complex directly from MGIT culture tubes. AB - Preservation of M. tuberculosis complex strains isolated from clinical specimens is important for epidemiological investigations related to tuberculosis. In this study the efficacy of preservation was evaluated by calculating the recovery rate of preserved strains, with various patterns of resistance, after periods of storage and subculture. The recovery rates from strains preserved in enriched solid medium were >90% for storage periods 90% of 'viable' hES cell clumps with such morphology adhered after a further 24 hr of culture on newly-plated MEF. The remaining non-adhered cell clumps that had ill-defined jagged edges could represent either non-viable hES cells or senescent MEF carried over from the previous sub-culture. Virtually none of these adhered to the newly plated MEF feeder layer. The overwhelming majority of 'viable,' late-adhering hES cell clumps (typically >90%) gave rise to morphologically 'normal-looking' hES colonies, which appeared to remain undifferentiated. Additionally, hES cell clumps that were attached together with the majority of the passaged colonies, but were somewhat delayed in the adhesion process, compared to neighbouring colonies, also gave rise to morphologically 'normal-looking' undifferentiated colonies that were virtually indistinguishable from neighbouring colonies. Hence, it may be useful to retain late-adhering hES cell clumps during serial passage of hES cells, rather than discarding them. PMID- 16254267 TI - Entries on laboratory medicine in the first illustrated medical dictionary. PMID- 16254268 TI - The beta-agonist lung injury trial (BALTI): a randomized placebo-controlled clinical trial. AB - RATIONALE: Experimental data suggest that manipulation of alveolar fluid clearance with beta-agonists can accelerate the resolution of alveolar edema and improve survival. OBJECTIVE: To determine if a sustained infusion of intravenous salbutamol (albuterol) would accelerate the resolution of alveolar edema in adult patients with acute lung injury (ALI) or acute respiratory distress syndrome (ARDS). METHODS: This was a single-center, double-blind, randomized controlled trial. Patients with ALI/ARDS were randomized to treatment with intravenous salbutamol (15 microg kg(-1) h(-1)) or placebo for 7 d. The primary endpoint was extravascular lung water measured by thermodilution (PiCCO) at Day 7. MEASUREMENTS AND MAIN RESULTS: Sixty-six patients were screened; of these, 40 met the inclusion criteria and were enrolled during 2001-2003. Patients in the salbutamol group had significantly lower lung water at Day 7 than the placebo group (9.2 +/- 6 vs. 13.2 +/- 3 ml kg(-1); 95% confidence interval difference, 0.2-8.3 ml kg(-1); p = 0.038). Plateau airway pressure was lower at Day 7 in the salbutamol group (23.9 +/- 3.8 cm H2O) versus placebo (29.5 +/- 7.2 cm H2O; p = 0.049). There was a trend toward lower Murray lung injury score at Day 7 in the salbutamol group (1.7 +/- 0.9) versus placebo (2.0 +/- 0.6; p = 0.2). Patients in the salbutamol group had a higher incidence of supraventricular arrhythmias (26 vs. 10%; p = 0.2). CONCLUSION: Although further research is required to confirm the efficacy and safety of intravenous salbutamol in ALI/ARDS, this trial provides the first proof of principle that, in humans with ALI/ARDS, sustained treatment with intravenous beta-agonists reduces extravascular lung water. PMID- 16254269 TI - Partial liquid ventilation in adult patients with acute respiratory distress syndrome. AB - RATIONALE: Despite recent clinical trials demonstrating improved outcome in acute respiratory distress syndrome (ARDS), mortality remains high. Partial liquid ventilation (PLV) using perfluorocarbons has been shown to improve oxygenation and decrease lung injury in various animal models. OBJECTIVE: To determine if PLV would have an impact on outcome in patients with ARDS. METHODS: Patients with ARDS were randomized to (1) conventional mechanical ventilation (CMV; n=107), (2) "low-dose" perfluorocarbon (10 ml/kg; n=99), and (3) "high-dose" perfluorocarbon (20 ml/kg; n=105). Patients in all three groups were ventilated using volume ventilation, Vt or= 0.5, and positive end expiratory pressure >or= 13 cm H(2)O. RESULTS: The 28-d mortality in the CMV group was 15%, versus 26.3% in the low-dose (p=0.06) and 19.1% in the high-dose (p=0.39) PLV groups. There were more ventilator-free days in the CMV group (13.0+/-9.3) compared with both the low-dose (7.4+/-8.5; p<0.001) and high-dose (9.9+/-9.1; p=0.043) groups. There were more pneumothoraces, hypoxic episodes, and hypotensive episodes in the PLV patients. CONCLUSIONS: PLV at both high and low doses did not improve outcome in ARDS compared with CMV and cannot be recommended for patients with ARDS. PMID- 16254270 TI - Association of indoor nitrogen dioxide exposure with respiratory symptoms in children with asthma. AB - RATIONALE: Chronic exposure to indoor nitrogen dioxide (NO2) is a public health concern. Over half of U.S. households have a source of NO2, and experimental data suggest potential for adverse respiratory effects. OBJECTIVE: To examine associations of indoor NO2 exposure with respiratory symptoms among children with asthma. METHODS: NO2 was measured using Palmes tubes, and respiratory symptoms in the month before sampling were collected during home interviews of mothers of 728 children with active asthma. All were younger than 12 yr, lived at the sampled home for at least 2 mo, and had asthma symptoms or used maintenance medication within the previous year. MEASUREMENTS: Respiratory symptoms (wheeze, persistent cough, shortness of breath, chest tightness). RESULTS: Mean (SD) NO2 was 8.6 (9.1) ppb in homes with electric ranges and 25.9 (18.1) ppb in homes with gas stoves. In models stratified by housing type (a factor associated with socioeconomic status), gas stove presence and elevated NO2 were each significantly associated with respiratory symptoms, controlling for age, ethnicity, medication, mold/mildew, water leaks, and season of sampling. Among children in multifamily housing, exposure to gas stoves increased likelihood of wheeze (odds ratio [OR], 2.27; 95% confidence interval [95% CI], 1.15, 4.47), shortness of breath (OR, 2.33; 95% CI, 1.12, 5.06), and chest tightness (OR, 4.34; 95% CI, 1.76, 10.69), whereas each 20-ppb increase in NO2 increased both likelihood of any wheeze (OR, 1.52; 95% CI, 1.04, 2.21) or chest tightness (OR, 1.61; 95% CI, 1.04, 2.49), and days of wheeze (rate ratio (RR), 1.33; 95% CI, 1.05, 1.68) or chest tightness (RR, 1.51; 95% CI, 1.18, 1.91). CONCLUSION: Exposure to indoor NO2 at levels well below the Environmental Protection Agency outdoor standard (53 ppb) is associated with respiratory symptoms among children with asthma in multifamily housing. PMID- 16254271 TI - Estimation of cancer mortality associated with repetitive computed tomography scanning. AB - RATIONALE: Low-dose radiation from computed tomography (CT) may increase the risk of certain cancers, especially in children. OBJECTIVE: We sought to estimate the excess all-cause and cancer-specific mortality, which may be associated with repeated CT scanning of patients with cystic fibrosis (CF). METHODS: The radiation dose was calculated for a published CF surveillance CT scanning protocol of biennial CT scans, and the risk per scan was estimated using atom bomb survivor data. A computational model was developed to calculate the excess mortality in a CF cohort associated with radiation from the CT scan and to evaluate the effects of background survival, scanning interval, and level of CT radiation used. The model assumed that there would be no survival benefits associated with repeated surveillance CT scanning. RESULTS: The average radiation dose for the published CT protocol was 1 mSv. Survival reduction associated with annual scans from age 2 yr until death was approximately 1 mo and 2 yr for CF cohorts, with a median survival of 26 and 50 yr, respectively. Corresponding cumulative cancer mortality was approximately 2 and 13% at age 40 and 65 yr, respectively. Biennial CT scanning reduced all-cause and cumulative cancer mortality by half. CONCLUSION: Routine lifelong annual CT scans carry a low risk of radiation-induced mortality in CF. However, as the overall survival increases for patients with CF, the risk of radiation-induced mortality may modestly increase. These data indicate that radiation dose must be considered in routine CT imaging strategies for patients with CF, to ensure that benefits outweigh the risks. PMID- 16254272 TI - Tracheobronchomalacia and air trapping after mustard gas exposure. AB - RATIONALE: Mustard gas primarily affects the eyes, skin, and particularly the respiratory tract. Tracheobronchomalacia (TBM) and air trapping are often observed in high-resolution computerized tomography (HRCT) scans of the chest of mustard gas-exposed patients. OBJECTIVES: To examine the frequency and severity of TBM in a group of Iranian wartime mustard gas-exposed victims, and to investigate the correlation between TBM and air trapping in these cases. MATERIALS AND METHODS: Chest HRCT films obtained from 300 randomly selected subjects who had been exposed to mustard gas 15.5 yr previously were reviewed to determine the existence of TBM and air trapping. The HRCT films of a healthy control group were also analyzed for comparison. RESULTS: Out of 300 reviewed cases, 13 had TBM. From these 13 TBM cases, 11 (85%) showed air trapping with mean score of 5.5. In the control group, 5 (25%) of 20 subjects showed air trapping, with mean score of 0.6. The total air trapping was significantly higher in the TBM group (p < 0.001). There was an association between the severity of tracheomalacia and air trapping in the TBM group (p = 0.01, r = 0.69), but no association was observed between severity of bronchomalacia and air trapping. CONCLUSION: The results show that air trapping and TBM are correlated, both as long-term sequelae in mustard gas-exposed cases. Because air trapping is highly suggestive of bronchiolitis obliterans, we conclude that both bronchiolitis obliterans and TBM are caused by a single underlying process affecting small and large airways, respectively, in this group of patients. PMID- 16254273 TI - Reactive species mediate inhibition of alveolar type II sodium transport during mycoplasma infection. AB - RATIONALE: Mycoplasma pneumoniae is a significant cause of pneumonia in humans. OBJECTIVES: To determine the impact of mycoplasma infection and the host inflammatory response on alveolar type II (ATII) cell ion transport in vivo and in vitro. METHODS: Mice were infected with M. pulmonis for measurements of alveolar fluid clearance (AFC) in vivo and isolation of ATII cells. ATII cells were infected in vivo for determination of epithelial Na+ channel (ENaC) total and cell surface protein levels by biotinylation and Western blot and in vitro for whole cell patch clamp recording and measurement of nitric oxide (NO) production by chemiluminescence. RESULTS: Mycoplasma infection significantly inhibited AFC at 24 h and total and amiloride-sensitive AFC by 48 h postinfection (pi). In contrast, infected myeloperoxidase-deficient mice had similar basal and amiloride-sensitive AFC values to uninfected control mice at 48 h pi. Addition of forskolin restored total and amiloride-sensitive AFC to control values at 48 h pi. ATII cells isolated from infected mice demonstrated normal alpha, beta, and gamma ENaC total protein levels; however, infected whole-lung cell-surface levels of gamma ENaC were significantly decreased. Patch-clamp recordings demonstrated a significant decrease in total and amiloride-sensitive Na+ currents at 24 h pi. ATII cells demonstrated a significant increase in the production of NO at 24 h pi and inhibition of NO by ATII cells before infection reversed the decrease in total Na+ currents. CONCLUSIONS: These data indicate that mycoplasma infection results in decreased AFC and functional ENaC via the production of reactive oxygen nitrogen intermediates. PMID- 16254274 TI - HLA-C and killer cell immunoglobulin-like receptor genes in idiopathic bronchiectasis. AB - RATIONALE: In idiopathic bronchiectasis, lung inflammation and chronic bacterial infection lead to progressive lung damage. A possible role for natural killer (NK) cells is suggested by the observation that familial bronchiectasis occurs in a rare group of individuals with impaired HLA class I expression and consequent NK cell dysfunction. OBJECTIVE: Because the HLA-C locus and killer cell immunoglobulin-like receptors (KIRs) are of key importance for NK cell recognition, we analyzed HLA-C/KIR combinations by genotyping patients with idiopathic bronchiectasis. METHODS: Genomic DNA from 96 individuals with idiopathic bronchiectasis and 101 control subjects was analyzed by polymerase chain reaction with sequence-specific primers. High-resolution HLA-C genotyping was performed in addition to KIR analysis. RESULTS: HLA-Cw*03 alleles and, in particular, HLA-C group 1 homozygosity are associated with the presence of bronchiectasis. Analysis of the relationship between HLA-C and KIR genes suggests a shift to activatory NK cell function. CONCLUSION: This is the first demonstration of genetic susceptibility in idiopathic bronchiectasis. The association with HLA-C group 1 homozygosity, and the interplay between HLA-C and KIR genes, argue for a role for NK cells in the progressive lung damage seen in this disease. This will require further investigation using functional studies. PMID- 16254275 TI - Improving surveillance of MRSA bacteraemia. PMID- 16254276 TI - The need for outcome measures in medical education. PMID- 16254277 TI - Detecting fabricated or induced illness in children. PMID- 16254278 TI - Near patient tests for smoking cessation. PMID- 16254279 TI - Bird flu poses no immediate threat to Europe, leading virologist claims. PMID- 16254281 TI - Stem cell foundation greeted by cautious optimism. PMID- 16254280 TI - Roche considers licensing companies to make oseltamivir. PMID- 16254284 TI - Patients with aortic aneurysm deserve higher priority. PMID- 16254290 TI - Judge over-rules earlier decision on Charlotte Wyatt. PMID- 16254292 TI - Total smoking ban best way to narrow health gap between rich and poor, MPs told. PMID- 16254294 TI - Health experts gather to step up fight against malaria. PMID- 16254298 TI - Charity calls for improved breast awareness in UK ethnic communities. PMID- 16254301 TI - Combating the free movement of micro-organisms. PMID- 16254300 TI - Experts disagree over who should get avian influenza vaccine. PMID- 16254302 TI - The patient's journey: the progressive ataxias. PMID- 16254303 TI - How computers help make efficient use of consultations. PMID- 16254304 TI - Problems in assessing rates of infection with methicillin resistant Staphylococcus aureus. PMID- 16254305 TI - Orphan drugs and the NHS: should we value rarity? PMID- 16254306 TI - Commissioning for rare diseases: view from the frontline. PMID- 16254307 TI - ASCOT: a tale of two treatment regimens: odds are that it's hype. PMID- 16254308 TI - ASCOT: a tale of two treatment regimens: is ASCOT-BPLA being hyped-up? PMID- 16254309 TI - ASCOT: a tale of two treatment regimens: don't lose your head over ASCOT. PMID- 16254310 TI - Left handedness and breast cancer: correlation is spurious. PMID- 16254311 TI - Trastuzumab for early breast cancer raises important issues. PMID- 16254312 TI - ASCOT: a tale of two treatment regimens: is ASCOT all it's cracked up to be? PMID- 16254313 TI - Scientific evidence was ignored in CAM and the NHS. PMID- 16254314 TI - Left handedness and breast cancer: should left handed premenopausal women be screened? PMID- 16254315 TI - Speed networking works for postgraduate research. PMID- 16254316 TI - Proposed guidelines for severe imported malaria in children need more evidence. PMID- 16254317 TI - Disappointing article, not disappointing biotech. PMID- 16254318 TI - Identification of quantitative trait loci that regulate obesity and serum lipid levels in MRL/MpJ x SJL/J inbred mice. AB - The total body fat mass and serum concentration of total cholesterol, HDL cholesterol, and triglyceride (TG) differ between standard diet-fed female inbred mouse strains MRL/MpJ (MRL) and SJL/J (SJL) by 38-120% (P < 0.01). To investigate genetic regulation of obesity and serum lipid levels, we performed a genome-wide linkage analysis in 621 MRLx SJL F2 female mice. Fat mass was affected by two significant loci, D11Mit36 [43.7 cM, logarithm of the odds ratio (LOD) 11.2] and D16Mit51 (50.3 cM, LOD 3.9), and one suggestive locus at D7Mit44 (50 cM, LOD 2.4). TG levels were affected by two novel loci at D1Mit43 (76 cM, LOD 3.8) and D12Mit201 (26 cM, LOD 4.1), and two suggestive loci on chromosomes 5 and 17. HDL and cholesterol concentrations were influenced by significant loci on chromosomes 1, 3, 5, 7, and 17 that were in the regions identified earlier for other strains of mice, except for a suggestive locus on chromosome 14 that was specific to the MRL x SJL cross. In summary, linkage analysis in MRL x SJL F2 mice disclosed novel loci affecting TG, HDL, and fat mass, a measure of obesity. Knowledge of the genes in these quantitative trait loci will enhance our understanding of obesity and lipid metabolism. PMID- 16254319 TI - Redundant roles for nucleocapsid and matrix RNA-binding sequences in human immunodeficiency virus type 1 assembly. AB - RNA appears to be required for the assembly of retroviruses. This is likely due to binding of RNA by multiple Gags, which in turn organizes and stabilizes the Gag-Gag interactions that form the virion. While the nucleocapsid (NC) domain is the most conspicuous RNA-binding region of the human immunodeficiency virus type 1 (HIV-1) Gag polyprotein, we have previously shown that NC is not strictly required for efficient particle production. To determine if an RNA requirement for HIV-1 assembly exists, we analyzed virions produced by an NC deletion mutant for the presence of RNA. The results revealed that virions without NC still contained significant amounts of RNA. Since these packaged RNAs are probably incorporated by other RNA-binding sequences in Gag, an RNA-binding site in the matrix protein (MA) of Gag was mutated. While this mutation did not interfere with HIV-1 replication, a construct with both MA and NC mutations (MX/NX) failed to produce particles. The MX/NX mutant was rescued in trans by coassembly with several forms of Gag: wild-type Gag, either of the single-mutant Gags, or Gag truncations that contain MA or NC sequences. Addition of basic sequences to the MX/NX mutant partially restored particle production, consistent with a requirement for Gag-RNA binding in addition to Gag-Gag interactions. Together, these results support an RNA-binding requirement for Gag assembly, which relies on binding of RNA by MA or NC sequences to condense, organize, and stabilize the HIV-1 Gag-Gag interactions that form the virion. PMID- 16254320 TI - Assembly of severe acute respiratory syndrome coronavirus RNA packaging signal into virus-like particles is nucleocapsid dependent. AB - The severe acute respiratory syndrome coronavirus (SARS-CoV) was recently identified as the etiology of SARS. The virus particle consists of four structural proteins: spike (S), small envelope (E), membrane (M), and nucleocapsid (N). Recognition of a specific sequence, termed the packaging signal (PS), by a virus N protein is often the first step in the assembly of viral RNA, but the molecular mechanisms involved in the assembly of SARS-CoV RNA are not clear. In this study, Vero E6 cells were cotransfected with plasmids encoding the four structural proteins of SARS-CoV. This generated virus-like particles (VLPs) of SARS-CoV that can be partially purified on a discontinuous sucrose gradient from the culture medium. The VLPs bearing all four of the structural proteins have a density of about 1.132 g/cm(3). Western blot analysis of the culture medium from transfection experiments revealed that both E and M expressed alone could be released in sedimentable particles and that E and M proteins are likely to form VLPs when they are coexpressed. To examine the assembly of the viral genomic RNA, a plasmid representing the GFP-PS580 cDNA fragment encompassing the viral genomic RNA from nucleotides 19715 to 20294 inserted into the 3' noncoding region of the green fluorescent protein (GFP) gene was constructed and applied to the cotransfection experiments with the four structural proteins. The SARS-CoV VLPs thus produced were designated VLP(GFP-PS580). Expression of GFP was detected in Vero E6 cells infected with the VLP(GFP-PS580), indicating that GFP-PS580 RNA can be assembled into the VLPs. Nevertheless, when Vero E6 cells were infected with VLPs produced in the absence of the viral N protein, no green fluorescence was visualized. These results indicate that N protein has an essential role in the packaging of SARS-CoV RNA. A filter binding assay and competition analysis further demonstrated that the N-terminal and C-terminal regions of the SARS-CoV N protein each contain a binding activity specific to the viral RNA. Deletions that presumably disrupt the structure of the N-terminal domain diminished its RNA binding activity. The GFP-PS-containing SARS-CoV VLPs are powerful tools for investigating the tissue tropism and pathogenesis of SARS-CoV. PMID- 16254321 TI - Kinetic analyses of the surface-transmembrane disulfide bond isomerization controlled fusion activation pathway in Moloney murine leukemia virus. AB - The surface (SU) and transmembrane (TM) subunits of Moloney murine leukemia virus (Mo-MLV) Env are disulfide linked. The linking cysteine in SU is part of a conserved CXXC motif in which the other cysteine carries a free thiol. Recently, we showed that receptor binding activates its free thiol to isomerize the intersubunit disulfide bond into a disulfide within the motif instead (M. Wallin, M. Ekstrom and H. Garoff, EMBO J. 23:54-65, 2004). This facilitated SU dissociation and activation of TM for membrane fusion. The evidence was mainly based on the finding that alkylation of the CXXC-thiol prevented isomerization. This arrested membrane fusion, but the activity could be rescued by cleaving the intersubunit disulfide bond with dithiothreitol (DTT). Here, we demonstrate directly that receptor binding causes SU-TM disulfide bond isomerization in a subfraction of the viral Envs. The kinetics of the isomerization followed that of virus-cell membrane fusion. Arresting the fusion with lysophosphatidylcholine did not arrest isomerization, suggesting that isomerization precedes the hemifusion stage of fusion. Our earlier finding that native Env was not possible to alkylate but required isomerization induction by receptor binding intimated that alkylation trapped an intermediate form of Env. To further clarify this possibility, we analyzed the kinetics by which the alkylation-sensitive Env was generated during fusion. We found that it followed the fusion kinetics. In contrast, the release of fusion from alkylated, isomerization-blocked virus by DTT reduction of the SU-TM disulfide bond was much faster. These results suggest that the alkylation-sensitive form of Env is a true intermediate in the fusion activation pathway of Env. PMID- 16254322 TI - Transcriptional regulation of early transposon elements, an active family of mouse long terminal repeat retrotransposons. AB - While early transposon (ETn) endogenous retrovirus (ERV)-like elements are known to be active insertional mutagens in the mouse, little is known about their transcriptional regulation. ETns are transcribed during early mouse embryogenesis in embryonic stem (ES) and embryonic carcinoma (EC) cell lines. Despite their lack of coding potential, some ETns remain transposition competent through their use of reverse transcriptase encoded by a related group of ERVs-MusD elements. In this study, we have confirmed high expression levels of ETn and MusD elements in ES and EC cells and have demonstrated an increase in the copy number of ETnII elements in the EC P19 cell line. Using transient transfections, we have shown that ETnII and MusD LTRs are much more active as promoters in P19 cells than in NIH 3T3 cells, indicating that genomic context and methylation are not the only factors determining endogenous transcriptional activity of ETns. Three sites in the 5' part of the long terminal repeat (LTR) were demonstrated to bind Sp1 and Sp3 transcription factors and were found to be important for high LTR promoter activity in P19 cells, suggesting that as yet unidentified Sp binding partners are involved in the regulation of ETn activity in undifferentiated cells. Finally, we found multiple transcription start sites within the ETn LTR and have shown that the LTR retains significant promoter activity in the absence of its noncanonical TATA box. These findings lend insight into the transcriptional regulation of this family of mobile mouse retrotransposons. PMID- 16254323 TI - Stress-activated protein kinases are involved in coxsackievirus B3 viral progeny release. AB - Stress-activated protein kinases (SAPKs), consisting of c-Jun N-terminal kinase (JNK) and p38 mitogen-activated protein kinase (p38 MAPK), are activated upon various environmental stimuli, including viral infections. Cellular survival and death signaling events following coxsackievirus B3 (CVB3) infection have been studied in relationship to viral replication, but the role of SAPKs has not been scrutinized. In this study, we found that the phosphorylation of JNK1/2 and p38 MAPK was increased during active replication of CVB3 and that their phosphorylation was independent of CVB3-induced caspase activation or production of reactive oxygen species. The roles of these kinases in CVB3 infection were further evaluated using specific inhibitors: SP600125 for JNK1/2 and SB203580 for p38 MAPK. JNK1/2 inhibitors reduced CVB3-induced phosphorylation of activating transcription factor 2, and the p38 MAPK inhibitor reduced CVB3-induced phosphorylation of heat shock protein 27. Although inhibition of these kinases by specific inhibitors did not affect CVB3 viral protein synthesis, inhibition of p38 MAPK but not of JNK1/2 resulted in significant reduction of viral progeny release, suppression of CVB3-induced cell death, and blockage of CVB3-induced caspase-3 activation in infected cells. We conclude that SAPK pathways play critical roles in the life cycle of CVB3, particularly in viral progeny release. PMID- 16254324 TI - Viral protein VP4 is a target of human antibodies enhancing coxsackievirus B4- and B3-induced synthesis of alpha interferon. AB - Coxsackievirus B4 (CVB4)-induced production of alpha interferon (IFN-alpha) by peripheral blood mononuclear cells (PBMC) is enhanced in vitro by nonneutralizing anti-CVB4 antibodies from healthy subjects and, to a higher extent, from patients with insulin-dependent diabetes mellitus. In this study, we focused on identification of the viral target of these antibodies in CVB systems. High levels of IFN-alpha were obtained in supernatants of PBMC incubated with CVB4E2 or CVB3 and plasma from healthy subjects and, to a higher extent, from patients. The VP4 capsid proteins dissociated by heating at 56 degrees C from CVB4E2 (VP4(CVB4)) and CVB3 (VP4(CVB3)) but not H antigen preincubated with plasma from healthy subjects or patients inhibited the plasma-dependent enhancement of CVB4E2 and CVB3-induced IFN-alpha synthesis. There was no cross-reaction between VP4(CVB4) and VP4(CVB3) in the inhibiting effect. IFN-alpha levels in culture supernatants showed dose-dependent correlation with anti-VP4 antibodies eluted from plasma specimens using VP4-coated plates. There were higher index values for anti-VP4 antibodies detected by enzyme-linked immunosorbent assay (ELISA) and higher proportions of positive detection in 40 patients than in 40 healthy subjects (80% versus 15% for anti-VP4(CVB4)). There was no relationship between the levels of anti-CVB neutralizing antibodies and the detection of anti-VP4 antibodies by ELISA. The CVB plasma-induced IFN-alpha levels obtained in PBMC cultures in the anti-VP4 antibody-positive groups were significantly higher than those obtained in the anti-VP4 antibody-negative groups regardless of the titers of anti-CVB neutralizing antibodies. These results show that VP4 is the target of antibodies involved in the plasma-dependent enhancement of CVB4E2- and CVB3 induced IFN-alpha synthesis by PBMC. PMID- 16254325 TI - Inhibition of polyprotein processing and RNA replication of human rhinovirus by pyrrolidine dithiocarbamate involves metal ions. AB - Pyrrolidine dithiocarbamate (PDTC) is an antiviral compound that was shown to inhibit the replication of human rhinoviruses (HRVs), poliovirus, and influenza virus. To elucidate the mechanism of PDTC, the effects on the individual steps of the infection cycle of HRV were investigated. PDTC did not interfere with receptor binding or internalization by receptor mediated endocytosis of HRV2 particles into HeLa cells. But we demonstrate that the processing of the viral polyprotein was prevented by PDTC treatment in HeLa cells infected with HRV2. Furthermore, PDTC inhibited the replication of the viral RNA, even when added four hours post infection. As PDTC is described as a metal ion binding agent, we investigated the effect of other metal chelators on the multiplication of HRV2. We show that EDTA, omicron-phenanthroline, and bathocuproine disulfonic acid do not exhibit any antiviral properties. Surprisingly, these substances, coadministered with PDTC, abolished the antiviral effect of PDTC, suggesting that metal ions play a pivotal role in the inhibition of virus multiplication. These results suggest that PDTC inhibits the activity of the viral proteases in a metal ion dependent way. PMID- 16254326 TI - Expression and mutational analysis of Autographa californica nucleopolyhedrovirus HCF-1: functional requirements for cysteine residues. AB - The host cell-specific factor 1 gene (hcf-1) of the baculovirus Autographa californica multiple nucleopolyhedrovirus is required for efficient virus growth in TN368 cells but is dispensable for virus replication in SF21 cells. However, the mechanism of action of hcf-1 is unknown. To begin to understand its function in virus replication we have investigated the expression and localization pattern of HCF-1 in infected cells. Analysis of virus-infected TN368 cells showed that hcf-1 is expressed at an early time in the virus life cycle, between 2 and 12 h postinfection, and localized the protein to punctate nuclear foci. Through coprecipitation experiments we have confirmed that HCF-1 self-associates into dimers or higher-order structures. We also found that overexpression of HCF-1 repressed expression from the hcf-1 promoter in transient reporter assays. Mutagenesis of cysteine residues within a putative RING finger domain in the amino acid sequence of HCF-1 abolished self-association activity and suggests that the RING domain may be involved in this protein-protein interaction. A different but overlapping set of cysteine residues were required for efficient gene repression activity. Functional analysis of HCF-1 mutants showed that the cysteine amino acids required for both self-association and gene repression activities of HCF-1 were also required for efficient late-gene expression and occlusion body formation in TN368 cells. Mutational analysis also identified essential charged and hydrophobic amino acids located between two of the essential cysteine residues. We propose that HCF-1 is a RING finger-containing protein whose activity requires HCF-1 self-association and gene repression activity. PMID- 16254327 TI - Modulation of the immune response to the severe acute respiratory syndrome spike glycoprotein by gene-based and inactivated virus immunization. AB - Although the initial isolates of the severe acute respiratory syndrome (SARS) coronavirus (CoV) are sensitive to neutralization by antibodies through their spike (S) glycoprotein, variants of S have since been identified that are resistant to such inhibition. Optimal vaccine strategies would therefore make use of additional determinants of immune recognition, either through cellular or expanded, cross-reactive humoral immunity. Here, the cellular and humoral immune responses elicited by different combinations of gene-based and inactivated viral particles with various adjuvants have been assessed. The T-cell response was altered by different prime-boost immunizations, with the optimal CD8 immunity induced by DNA priming and replication-defective adenoviral vector boosting. The humoral immune response was enhanced most effectively through the use of inactivated virus with adjuvants, either MF59 or alum, and was associated with stimulation of the CD4 but not the CD8 response. The use of inactivated SARS virus with MF59 enhanced the CD4 and antibody response even after gene-based vaccination. Because both cellular and humoral immune responses are generated by gene-based vaccination and inactivated viral boosting, this strategy may prove useful in the generation of SARS-CoV vaccines. PMID- 16254328 TI - NS1 protein secretion during the acute phase of West Nile virus infection. AB - The West Nile virus (WNV) nonstructural protein NS1 is a protein of unknown function that is found within, associated with, and secreted from infected cells. We systematically investigated the kinetics of NS1 secretion in vitro and in vivo to determine the potential use of this protein as a diagnostic marker and to analyze NS1 secretion in relation to the infection cycle. A sensitive antigen capture enzyme-linked immunosorbent assay (ELISA) for detection of WNV NS1 (polyclonal-ACE) was developed, as well as a capture ELISA for the specific detection of NS1 multimers (4G4-ACE). The 4G4-ACE detected native NS1 antigens at high sensitivity, whereas the polyclonal-ACE had a higher specificity for recombinant forms of the protein. Applying these assays we found that only a small fraction of intracellular NS1 is secreted and that secretion of NS1 in tissue culture is delayed compared to the release of virus particles. In experimentally infected hamsters, NS1 was detected in the serum between days 3 and 8 postinfection, peaking on day 5, the day prior to the onset of clinical disease; immunoglobulin M (IgM) antibodies were detected at low levels on day 5 postinfection. Although real-time PCR gave the earliest indication of infection (day 1), the diagnostic performance of the 4G4-ACE was comparable to that of real time PCR during the time period when NS1 was secreted. Moreover, the 4G4-ACE was found to be superior in performance to both the IgM and plaque assays during this time period, suggesting that NS1 is a viable early diagnostic marker of WNV infection. PMID- 16254329 TI - A live attenuated vaccine for Lassa fever made by reassortment of Lassa and Mopeia viruses. AB - Lassa virus (LASV) and Mopeia virus (MOPV) are closely related Old World arenaviruses that can exchange genomic segments (reassort) during coinfection. Clone ML29, selected from a library of MOPV/LASV (MOP/LAS) reassortants, encodes the major antigens (nucleocapsid and glycoprotein) of LASV and the RNA polymerase and zinc-binding protein of MOPV. Replication of ML29 was attenuated in guinea pigs and nonhuman primates. In murine adoptive-transfer experiments, as little as 150 PFU of ML29 induced protective cell-mediated immunity. All strain 13 guinea pigs vaccinated with clone ML29 survived at least 70 days after LASV challenge without either disease signs or histological lesions. Rhesus macaques inoculated with clone ML29 developed primary virus-specific T cells capable of secreting gamma interferon in response to homologous MOP/LAS and heterologous MOPV and lymphocytic choriomeningitis virus. Detailed examination of two rhesus macaques infected with this MOPV/LAS reassortant revealed no histological lesions or disease signs. Thus, ML29 is a promising attenuated vaccine candidate for Lassa fever. PMID- 16254330 TI - Overexpression of the M2-2 protein of respiratory syncytial virus inhibits viral replication. AB - The M2-2 protein of respiratory syncytial virus (RSV) is involved in regulation of viral RNA transcription and replication. Encoded by the next-to-last gene of RSV, the M2-2 open reading frame (ORF) overlaps with the upstream M2-1 ORF, suggesting that the production of the M2-2 protein might be tightly regulated during virus replication. To evaluate the effect of M2-2 overexpression on RSV replication, the M2-2 gene was separated from M2-1 by leaving it at the position prior to the M2-1 or moving it to the promoter proximal position as an independent transcriptional unit in the RSV A2 genome. Although recombinant viruses bearing the shuffled M2-2 gene were recovered and expressed higher levels of M2-2, most of these viruses grew poorly in HEp-2 cells. Sequence analysis revealed that various mutations (substitution, insertion, and deletion) occurred in the M2-2 gene, resulting in reduced M2-2 activity as measured by the RSV minigenome system. Further examination of the M2-2 sequence and its function showed that either one of the first two AUG codons located at the 5' end of M2-2 could be used to produce a functional M2-2 protein and that deletion of the first six amino acids from its N terminus or four amino acids from its C terminus greatly reduced its function. The effect of M2-2 protein on RSV replication was also studied by examining RSV replication in cells transiently expressing M2-2. The M2-2 protein expressed at a high level completely inhibited RSV replication. These results strongly suggested that the level of the M2-2 protein produced in the infected cells is critical to RSV replication. PMID- 16254331 TI - Sexual transmission of single human immunodeficiency virus type 1 virions encoding highly polymorphic multisite cytotoxic T-lymphocyte escape variants. AB - Antigenic variation inherent in human immunodeficiency virus type 1 (HIV-1) virions that successfully instigate new infections transferred by sex has not been well defined. Yet this is the viral "challenge" which any vaccine-induced immunity must deal with. Closely timed comparisons of the virus circulating in the "donor" and that which initiates new infection are difficult to carry out rigorously, as suitable samples are very hard to get in the face of ethical hurdles. Here we investigate HIV-1 variation in four homosexual couples where we sampled blood from both parties within several weeks of the estimated transmission event. We analyzed variation within highly immunogenic HIV-1 internal proteins encoding epitopes recognized by cytotoxic T lymphocytes (CTLs). These responses are believed to be crucial as a means of containing viral replication. In the donors we detected virions capable of evading host CTL recognition at several linked epitopes of distinct HLA class I restriction. When a donor transmitted escape variants to a recipient with whom he had HLA class I molecules in common, the recipient's CTL response to those epitopes was prevented, thus impeding adequate viral control. In addition, we show that even when HLA class I alleles are disparate in the transmitting couple, a single polymorphism can abolish CTL recognition of an overlapping epitope of distinct restriction and so confer immune escape properties to the recipient's seroconversion virus. In donors who are themselves controlling an early, acute infection, the precise timing of onward transmission is a crucial determinant of the viral variants available to compose the inoculum. PMID- 16254332 TI - Robust production of infectious hepatitis C virus (HCV) from stably HCV cDNA transfected human hepatoma cells. AB - Hepatitis C virus (HCV) chronically infects approximately 170 million people worldwide, with an increased risk of developing cirrhosis and hepatocellular carcinoma. The study of HCV replication and pathogenesis has been hampered by the lack of an efficient stable cell culture system and small-animal models of HCV infection and propagation. In an effort to develop a robust HCV infection system, we constructed stable human hepatoma cell lines that contain a chromosomally integrated genotype 2a HCV cDNA and constitutively produce infectious virus. Transcriptional expression of the full-length HCV RNA genome is under the control of a cellular Pol II polymerase promoter at the 5' end and a hepatitis delta virus ribozyme at the 3' end. The resulting HCV RNA was expressed and replicated efficiently, as shown by the presence of high levels of HCV proteins as well as both positive- and negative-strand RNAs in the stable Huh7 cell lines. Stable cell lines robustly produce HCV virions with up to 10(8) copies of HCV viral RNA per milliliter (ml) of the culture medium. Subsequent infection of naive Huh7.5 cells with HCV released from the stable cell lines resulted in high levels of HCV proteins and RNAs. Additionally, HCV infection was inhibited by monoclonal antibodies specific to CD81 and the HCV envelope glycoproteins E1 and E2, and HCV replication was suppressed by alpha interferon. Collectively, these results demonstrate the establishment of a stable HCV culture system that robustly produces infectious virus, which will allow the study of each aspect of the entire HCV life cycle. PMID- 16254333 TI - Identification of interferon-stimulated gene 15 as an antiviral molecule during Sindbis virus infection in vivo. AB - The innate immune response, and in particular the alpha/beta interferon (IFN alpha/beta) system, plays a critical role in the control of viral infections. Interferons alpha and beta exert their antiviral effects through the induction of hundreds of interferon-induced (or -stimulated) genes (ISGs). While several of these ISGs have characterized antiviral functions, their actions alone do not explain all of the effects mediated by IFN-alpha/beta. To identify additional IFN induced antiviral molecules, we utilized a recombinant chimeric Sindbis virus to express selected ISGs in IFN-alpha/beta receptor (IFN-alpha/betaR)(-/-) mice and looked for attenuation of Sindbis virus infection. Using this approach, we identified a ubiquitin homolog, interferon-stimulated gene 15 (ISG15), as having antiviral activity. ISG15 expression protected against Sindbis virus-induced lethality and decreased Sindbis virus replication in multiple organs without inhibiting the spread of virus throughout the host. We establish that, much like ubiquitin, ISG15 requires its C-terminal LRLRGG motif to form intracellular conjugates. Finally, we demonstrate that ISG15's LRLRGG motif is also required for its antiviral activity. We conclude that ISG15 can be directly antiviral. PMID- 16254334 TI - Epstein-Barr virus lytic infection is required for efficient production of the angiogenesis factor vascular endothelial growth factor in lymphoblastoid cell lines. AB - Although Epstein-Barr virus (EBV)-associated malignancies are primarily composed of cells with one of the latent forms of EBV infection, a small subset of tumor cells containing the lytic form of infection is often observed. Whether the rare lytically infected tumor cells contribute to the growth of the latently infected tumor cells is unclear. Here we have investigated whether the lytically infected subset of early-passage lymphoblastoid cell lines (LCLs) could potentially contribute to tumor growth through the production of angiogenesis factors. We demonstrate that supernatants from early-passage LCLs infected with BZLF1-deleted virus (Z-KO LCLs) are highly impaired in promoting endothelial cell tube formation in vitro compared to wild-type (WT) LCL supernatants. Furthermore, expression of the BZLF1 gene product in trans in Z-KO LCLs restored angiogenic capacity. The supernatants of Z-KO LCLs, as well as supernatants from LCLs derived with a BRLF1-deleted virus (R-KO LCLs), contained much less vascular endothelial growth factor (VEGF) in comparison to WT LCLs. BZLF1 gene expression in Z-KO LCLs restored the VEGF level in the supernatant. However, the cellular level of VEGF mRNA was similar in Z-KO, R-KO, and WT LCLs, suggesting that lytic infection may enhance VEGF translation or secretion. Interestingly, a portion of the vasculature in LCL tumors in SCID mice was derived from the human LCLs. These results suggest that lytically infected cells may contribute to the growth of EBV associated malignancies by enhancing angiogenesis. In addition, as VEGF is a pleiotropic factor with effects other than angiogenesis, lytically induced VEGF secretion may potentially contribute to viral pathogenesis. PMID- 16254335 TI - Epstein-Barr virus lytic infection contributes to lymphoproliferative disease in a SCID mouse model. AB - Most Epstein-Barr virus (EBV)-positive tumor cells contain one of the latent forms of viral infection. The role of lytic viral gene expression in EBV associated malignancies is unknown. Here we show that EBV mutants that cannot undergo lytic viral replication are defective in promoting EBV-mediated lymphoproliferative disease (LPD). Early-passage lymphoblastoid cell lines (LCLs) derived from EBV mutants with a deletion of either viral immediate-early gene grew similarly to wild-type (WT) virus LCLs in vitro but were deficient in producing LPD when inoculated into SCID mice. Restoration of lytic EBV gene expression enhanced growth in SCID mice. Acyclovir, which prevents lytic viral replication but not expression of early lytic viral genes, did not inhibit the growth of WT LCLs in SCID mice. Early-passage LCLs derived from the lytic defective viruses had substantially decreased expression of the cytokine interleukin-6 (IL-6), and restoration of lytic gene expression reversed this defect. Expression of cellular IL-10 and viral IL-10 was also diminished in lytic defective LCLs. These results suggest that lytic EBV gene expression contributes to EBV-associated lymphoproliferative disease, potentially through induction of paracrine B-cell growth factors. PMID- 16254336 TI - Adenovirus exploits the cellular aggresome response to accelerate inactivation of the MRN complex. AB - Results reported here indicate that adenovirus 5 exploits the cellular aggresome response to accelerate inactivation of MRE11-RAD50-NBS1 (MRN) complexes that otherwise inhibit viral DNA replication and packaging. Aggresomes are cytoplasmic inclusion bodies, observed in many degenerative diseases, that are formed from aggregated proteins by dynein-dependent retrograde transport on microtubules to the microtubule organizing center. Viral E1B-55K protein forms aggresomes that sequester p53 and MRN in transformed cells and in cells transfected with an E1B 55K expression vector. During adenovirus infection, the viral protein E4orf3 associates with MRN in promyelocytic leukemia protein nuclear bodies before MRN is bound by E1B-55K. Either E4orf3 or E4orf6 is required in addition to E1B-55K for E1B-55K aggresome formation and MRE11 export to aggresomes in adenovirus infected cells. Aggresome formation contributes to the protection of viral DNA from MRN activity by sequestering MRN in the cytoplasm and greatly accelerating its degradation by proteosomes following its ubiquitination by the E1B 55K/E4orf6/elongin BC/Cullin5/Rbx1 ubiquitin ligase. Our results show that aggresomes significantly accelerate protein degradation by the ubiquitin proteosome system. The observation that a normal cellular protein is inactivated when sequestered into an aggresome through association with an aggresome-inducing protein has implications for the potential cytotoxicity of aggresome-like inclusion bodies in degenerative diseases. PMID- 16254337 TI - The p domain of norovirus capsid protein forms a subviral particle that binds to histo-blood group antigen receptors. AB - Norovirus is the most important cause of nonbacterial acute gastroenteritis. We have shown previously that the isolated P domain containing the hinge forms a dimer and binds to histo-blood group antigen (HBGA) receptors with a low affinity (M. Tan, R. S. Hegde, and X. Jiang, J. Virol. 78:6233-6242, 2004). Here, we reported that the P domain of VA387 without the hinge forms a small particle with a significantly increased receptor binding affinity. An end-linked oligopeptide containing one or more cysteines promoted P-particle formation by forming intermolecular disulfide bridges. The binding sensitivity of the P particle to HBGAs was enhanced >700-fold compared to the P dimer, which was comparable to that of virus-like particles. The binding specificity of the P particle was further confirmed by strong binding to the Caco-2 cells, a human colon carcinoma cell line. This binding enhancement was observed in the P particles of both norovirus GI and GII strains. The P particle is estimated to contain 12 P dimers, in which the P2 subdomain builds up the outer layer, while the P1 subdomain forms the internal core. Taken together, our data indicate that the P domain is involved not only in dimerization but also in polymerization of the protein during the capsid assembling. The enhanced receptor binding of the P particle reflects the intrinsic feature of the viral capsid. The easy production of the P particle and its strong binding to HBGAs suggest that the P particle is useful in studying pathogenesis and morphogenesis of norovirus and candidates for antiviral or vaccine development. PMID- 16254338 TI - The vaccinia virus F1L protein interacts with the proapoptotic protein Bak and inhibits Bak activation. AB - Many viruses have evolved strategies to counteract cellular immune responses, including apoptosis. Vaccinia virus, a member of the poxvirus family, encodes an antiapoptotic protein, F1L. F1L localizes to mitochondria and inhibits apoptosis by preventing the release of cytochrome c by an undetermined mechanism (S. T. Wasilenko, T. L. Stewart, A. F. Meyers, and M. Barry, Proc. Natl. Acad. Sci. USA 100:14345-14350, 2003; T. L. Stewart, S. T. Wasilenko, and M. Barry, J. Virol. 79:1084-1098, 2005). Here, we show that in the absence of an apoptotic stimulus, F1L associates with Bak, a proapoptotic member of the Bcl-2 family that plays a pivotal role in the release of cytochrome c. Cells infected with vaccinia virus were resistant to Bak oligomerization and the initial N-terminal exposure of Bak following the induction of apoptosis with staurosporine. A mutant vaccinia virus missing F1L was no longer able to inhibit apoptosis or Bak activation. In addition, the expression of F1L was essential to inhibit tBid-induced cytochrome c release in both wild-type murine embryonic fibroblasts (MEFs) and Bax-deficient MEFs, indicating that F1L could inhibit apoptosis in the presence and absence of Bax. tBid-induced Bak oligomerization and N-terminal exposure of Bak in Bax deficient MEFs were inhibited during virus infection, as assessed by cross linking and limited trypsin proteolysis. Infection with the F1L deletion virus no longer provided protection from tBid-induced Bak activation and apoptosis. Additionally, infection of Jurkat cells with the F1L deletion virus resulted in cellular apoptosis, as measured by loss of the inner mitochondrial membrane potential, caspase 3 activation, and cytochrome c release, indicating that the presence of F1L was pivotal for inhibiting vaccinia virus-induced apoptosis. Our data indicate that F1L expression during infection inhibits apoptosis and interferes with the activation of Bak. PMID- 16254339 TI - Immunodeficiency in the absence of high viral load in pig-tailed macaques infected with Simian immunodeficiency virus SIVsun or SIVlhoest. AB - Simian immunodeficiency virus (SIV) is known to result in an asymptomatic infection of its natural African monkey host. However, some SIV strains are capable of inducing AIDS-like symptoms and death upon experimental infection of Asian macaques. To further investigate the virulence of natural SIV isolates from African monkeys, pig-tailed (PT) macaques were inoculated intravenously with either of two recently discovered novel lentiviruses, SIVlhoest and SIVsun. Both viruses were apparently apathogenic in their natural hosts but caused immunodeficiency in PT macaques. Infection was characterized by a progressive loss of CD4(+) lymphocytes in the peripheral blood and lymph nodes, generalized lymphoid depletion, a wasting syndrome, and opportunistic infections, such as Mycobacterium avium or Pneumocystis carinii infections. However, unlike SIVsm/mac infection of macaques, SIVlhoest and SIVsun infections in PT macaques were not accompanied by high viral loads during the chronic disease stage. In addition, no significant correlation between the viral load at set point (12 weeks postinfection) and survival could be found. Five out of eight SIVlhoest-infected and three out of four SIVsun-infected macaques succumbed to AIDS during the first 5 years of infection. Thus, the survival of SIVsun- and SIVlhoest-infected animals was significantly longer than that of SIVagm- or SIVsm-infected macaques. All PT macaques maintained strong SIV antibody responses despite progression to SIV-induced AIDS. The development of immunodeficiency in the face of low viremia suggests that SIVlhoest and SIVsun infections of macaques may model unique aspects of the pathogenesis of human immunodeficiency virus infection in humans. PMID- 16254340 TI - Phosphorylation of the herpes simplex virus tegument protein VP22 has no effect on incorporation of VP22 into the virus but is involved in optimal expression and virion packaging of ICP0. AB - Herpes simplex virus VP22 is a major tegument protein of unknown function. Very recently, we reported that the predominant effect of deleting the VP22 gene was on the expression, localization, and virion incorporation of ICP0. In addition, the Delta22 virus replicated poorly in epithelial MDBK cells. We have also previously shown that VP22 interacts with the tegument protein VP16 and the cellular microtubule network. While the majority of VP22 in infected cells is highly phosphorylated, the nonphosphorylated form of VP22 is the predominant species in the virion, suggesting a differential requirement for phosphorylation through virus replication. Hence, to study the significance of VP22 phosphorylation, we have now constructed two recombinant viruses expressing green fluorescent protein-VP22 (G22) in which the previously identified serine phosphorylation sites have been mutated either to alanine to abolish the phosphorylation status of VP22 (G22P-) or to glutamic acid to mimic permanent phosphorylation (G22P+). Localization studies indicated that the G22P- protein associated tightly with microtubules in some infected cells, suggesting that VP22 phosphorylation may control its interaction with the microtubule network. By contrast, VP22 phosphorylation had no effect on its ability to interact with VP16 and, importantly, had no effect on the relative packaging of VP22. Intriguingly, virion packaging of ICP0 was reduced in the G22P+ virus while ICP0 expression was reduced in the G22P- virus, suggesting that these two ICP0 defects, previously observed in the Delta22 virus, were attributable to different forms of VP22. Furthermore, the Delta22 virus replication defect in MDBK cells correlated with the expression of constitutively charged VP22 in the G22P+ virus. Taken together, these results suggest an important role for VP22 phosphorylation in its relationship with ICP0. PMID- 16254342 TI - Laser-capture microdissection: refining estimates of the quantity and distribution of latent herpes simplex virus 1 and varicella-zoster virus DNA in human trigeminal Ganglia at the single-cell level. AB - There remains uncertainty and some controversy about the percentages and types of cells in human sensory nerve ganglia that harbor latent herpes simplex virus 1 (HSV-1) and varicella-zoster virus (VZV) DNA. We developed and validated laser capture microdissection and real-time PCR (LCM/PCR) assays for the presence and copy numbers of HSV-1 gG and VZV gene 62 sequences in single cells recovered from sections of human trigeminal ganglia (TG) obtained at autopsy. Among 970 individual sensory neurons from five subjects, 2.0 to 10.5% were positive for HSV 1 DNA, with a median of 11.3 copies/positive cell, compared with 0.2 to 1.5% of neurons found to be positive by in situ hybridization (ISH) for HSV-1 latency associated transcripts (LAT), the classical surrogate marker for HSV latency. This indicates a more pervasive latent HSV-1 infection of human TG neurons than originally thought. Combined ISH/LCM/PCR assays revealed that the majority of the latently infected neurons do not accumulate LAT to detectable levels. We detected VZV DNA in 1.0 to 6.9% of individual neurons from 10 subjects. Of the total 1,722 neurons tested, 4.1% were VZV DNA positive, with a median of 6.9 viral genomes/positive cell. After removal by LCM of all visible neurons on a slide, all surrounding nonneuronal cells were harvested and assayed: 21 copies of HSV-1 DNA were detected in approximately 5,200 nonneuronal cells, while nine VZV genomes were detected in approximately 14,200 nonneuronal cells. These data indicate that both HSV-1 and VZV DNAs persist in human TG primarily, if not exclusively, in a moderate percentage of neuronal cells. PMID- 16254341 TI - Induction of cell cycle arrest by human T-cell lymphotropic virus type 1 Tax in hematopoietic progenitor (CD34+) cells: modulation of p21cip1/waf1 and p27kip1 expression. AB - Human T-cell lymphotropic virus type 1 (HTLV-1) is the etiologic agent of adult T cell leukemia, an aggressive CD4(+) malignancy. Although HTLV-2 is highly homologous to HTLV-1, infection with HTLV-2 has not been associated with lymphoproliferative disorders. Lentivirus-mediated transduction of CD34(+) cells with HTLV-1 Tax (Tax1) induced G(0)/G(1) cell cycle arrest and resulted in the concomitant suppression of multilineage hematopoiesis in vitro. Tax1 induced transcriptional upregulation of the cdk inhibitors p21(cip1/waf1) (p21) and p27(kip1) (p27), and marked suppression of hematopoiesis in immature (CD34(+)/CD38(-)) hematopoietic progenitor cells in comparison to CD34(+)/CD38(+) cells. HTLV-1 infection of CD34(+) cells also induced p21 and p27 expression. Tax1 also protected CD34(+) cells from serum withdrawal-mediated apoptosis. In contrast, HTLV-2 Tax (Tax2) did not detectably alter p21 or p27 gene expression, failed to induce cell cycle arrest, failed to suppress hematopoiesis in CD34(+) cells, and did not protect cells from programmed cell death. A Tax2/Tax1 chimera encoding the C-terminal 53 amino acids of Tax1 fused to Tax2 (Tax(221)) displayed a phenotype in CD34(+) cells similar to that of Tax1, suggesting that unique domains encoded within the C terminus of Tax1 may account for the phenotypes displayed in human hematopoietic progenitor cells. These remarkable differences in the activities of Tax1 and Tax2 in CD34(+) hematopoietic progenitor cells may underlie the sharp differences observed in the pathogenesis resulting from infection with HTLV-1 and HTLV-2. PMID- 16254343 TI - Crystal structure of enteric adenovirus serotype 41 short fiber head. AB - Human enteric adenoviruses of species F contain two fibers in the same virion, a long fiber which binds to coxsackievirus and adenovirus receptor (CAR) and a short fiber of unknown function. We have determined the high-resolution crystal structure of the short fiber head of human adenovirus serotype 41 (Ad41). The short fiber head has the characteristic fold of other known fiber heads but has three unusual features. First, it has much shorter loops between the beta strands. Second, one of the usually well-ordered beta-strands on the distal face of the fiber head is highly disordered and this same region is sensitive to digestion with pepsin, an enzyme occurring naturally in the intestinal tract, the physiological environment of Ad41. Third, the AB loop has a deletion giving it a distinct conformation incompatible with CAR binding. PMID- 16254344 TI - Gene and genome duplication in Acanthamoeba polyphaga Mimivirus. AB - Gene duplication is key to molecular evolution in all three domains of life and may be the first step in the emergence of new gene function. It is a well recognized feature in large DNA viruses but has not been studied extensively in the largest known virus to date, the recently discovered Acanthamoeba polyphaga Mimivirus. Here, I present a systematic analysis of gene and genome duplication events in the mimivirus genome. I found that one-third of the mimivirus genes are related to at least one other gene in the mimivirus genome, either through a large segmental genome duplication event that occurred in the more remote past or through more recent gene duplication events, which often occur in tandem. This shows that gene and genome duplication played a major role in shaping the mimivirus genome. Using multiple alignments, together with remote-homology detection methods based on Hidden Markov Model comparison, I assign putative functions to some of the paralogous gene families. I suggest that a large part of the duplicated mimivirus gene families are likely to interfere with important host cell processes, such as transcription control, protein degradation, and cell regulatory processes. My findings support the view that large DNA viruses are complex evolving organisms, possibly deeply rooted within the tree of life, and oppose the paradigm that viral evolution is dominated by lateral gene acquisition, at least in regard to large DNA viruses. PMID- 16254345 TI - Epstein-Barr virus mRNA export factor EB2 is essential for intranuclear capsid assembly and production of gp350. AB - Most human herpesviruses, including Epstein-Barr virus (EBV), express a protein which functions primarily as an mRNA export factor. Previously, we deleted the gene for the Epstein-Barr virus mRNA export factor EB2 from the EBV genome and then introduced the mutated genome into 293 cells. Using a transcomplementation assay in which ectopic expression of the transcription factor EB1/ZEBRA was sufficient to induce the EBV productive cycle, we showed that Ori-Lyt-dependent replication of the EBV DNA occurs in the absence of EB2, indicating that EB2 is not essential for the expression and export of early mRNAs. However, in the absence of EB2, no infectious viral particles are produced (H. Gruffat, J. Batisse, D. Pich, B. Neuhierl, E. Manet, W. Hammerschmidt, and A. Sergeant, J. Virol. 76:9635-9644, 2002). In this report, we now show that EB2 is essential for the nuclear export of most, but not all, late mRNAs produced from intronless genes that translate into proteins involved in intranuclear capsid assembly and maturation. As a consequence, we show that EB2 is essential for the proper assembly of intranuclear capsids. Interestingly, the late BLLF1 gene contains an intron, and both unspliced and spliced mRNAs must be exported to the cytoplasm to be translated into gp350 and gp220, respectively (M. Hummel, D. A. Thorley Lawson, and E. Kieff, J. Virol. 49:413-417, 1984). Our results also demonstrate that although BLLF1 spliced mRNAs are exported to the cytoplasm independently of EB2, EB2 is essential for the nuclear export of unspliced BLLF1 mRNA. In the same assay, herpes simplex virus 1 ICP27 completely inhibited the nuclear export of BLLF1 spliced mRNAs whereas unspliced BLLF1 mRNAs were exported, confirming that in a physiological assay, ICP27 inhibits splicing. PMID- 16254346 TI - Interferon-induced alterations in the pattern of parainfluenza virus 5 transcription and protein synthesis and the induction of virus inclusion bodies. AB - Although parainfluenza virus 5 (simian virus 5 [SV5]) circumvents the interferon (IFN) response by blocking IFN signaling and by reducing the amount of IFN released by infected cells, its ability to circumvent the IFN response is not absolute. The effects of IFN on SV5 infection were examined in Vero cells, which do not produce but can respond to IFN, using a strain of SV5 (CPI-) which does not block IFN signaling. Thus, by infecting Vero cells with CPI- and subsequently treating the cells with exogenous IFN, it was possible to observe the effects that IFN had on SV5 infection in the absence of virus countermeasures. IFN rapidly (within 6 h) induced alterations in the relative levels of virus mRNA and protein synthesis and caused a redistribution of virus proteins within infected cells that led to the enhanced formation of virus cytoplasmic inclusion bodies. IFN induced a steeper gradient of mRNA transcription from the 3' to the 5' end of the genome and the production of virus mRNAs with longer poly(A) tails, suggesting that the processivity of the virus polymerase was altered in cells in an IFN-induced antiviral state. Additional evidence is presented which suggests that these findings also apply to the replication of strains of SV5, parainfluenza virus type 2, and mumps virus that block IFN signaling when they infect cells that are already in an IFN-induced antiviral state. PMID- 16254347 TI - Acquisition of macrophage tropism during the pathogenesis of feline infectious peritonitis is determined by mutations in the feline coronavirus spike protein. AB - In feline coronavirus (FCoV) pathogenesis, the ability to infect macrophages is an essential virulence factor. Whereas the low-virulence feline enteric coronavirus (FECV) isolates primarily replicate in the epithelial cells of the enteric tract, highly virulent feline infectious peritonitis virus (FIPV) isolates have acquired the ability to replicate efficiently in macrophages, which allows rapid dissemination of the virulent virus throughout the body. FIPV 79 1146 and FECV 79-1683 are two genetically closely related representatives of the two pathotypes. Whereas FECV 79-1683 causes at the most a mild enteritis in young kittens, FIPV 79-1146 almost invariably induces a lethal peritonitis. The virulence phenotypes correlate with the abilities of these viruses to infect and replicate in macrophages, a feature of FIPV 79-1146 but not of FECV 79-1683. To identify the genetic determinants of the FIPV 79-1146 macrophage tropism, we exchanged regions of its genome with the corresponding parts of FECV 79-1683, after which the ability of the FIPV/FECV hybrid viruses to infect macrophages was tested. Thus, we established that the FIPV spike protein is the determinant for efficient macrophage infection. Interestingly, this property mapped to the C terminal domain of the protein, implying that the difference in infection efficiency between the two viruses is not determined at the level of receptor usage, which we confirmed by showing that infection by both viruses was equally blocked by antibodies directed against the feline aminopeptidase N receptor. The implications of these findings are discussed. PMID- 16254349 TI - A single amino acid change in rabies virus glycoprotein increases virus spread and enhances virus pathogenicity. AB - Several rabies virus (RV) vaccine strains containing an aspartic acid (Asp) or glutamic acid (Glu) instead of an arginine (Arg) at position 333 of the RV glycoprotein (G) are apathogenic for immunocompetent mice even after intracranial inoculation. However, we previously showed that the nonpathogenic phenotype of the highly attenuated RV strain SPBNGA, which contains a Glu at position 333 of G, is unstable when this virus is passaged in newborn mice. While the Glu(333) remained unchanged after five mouse passages, an Asn(194)-->Lys(194) mutation occurred in RV G. This mutation was associated with increased pathogenicity for adult mice. Using site-directed mutagenesis to exchange Asn(194) with Lys(194) in the G protein of SPBNGA, resulting in SPBNGA-K, we show here that this mutation is solely responsible for the increase in pathogenicity and that the Asn(194)- >Lys(194) mutation does not arise when Asn(194) is exchanged with Ser(194) (SPBNGA-S). Our data presented indicate that the increased pathogenicity of SPBNGA-K is due to increased viral spread in vivo and in vitro, faster internalization of the pathogenic virus into cells, and a shift in the pH threshold for membrane fusion. These results are consistent with the notion that the RV G protein is a major contributor to RV pathogenesis and that the more pathogenic RVs escape the host responses by a faster spread than that of less pathogenic RVs. PMID- 16254348 TI - Association of human immunodeficiency virus type 1 gag with membrane does not require highly basic sequences in the nucleocapsid: use of a novel Gag multimerization assay. AB - Human immunodeficiency virus type 1 (HIV-1) particle production, a process driven by the Gag polyprotein precursor, occurs on the plasma membrane in most cell types. The plasma membrane contains cholesterol-enriched microdomains termed lipid rafts, which can be isolated as detergent-resistant membrane (DRM). Previously, we and others demonstrated that HIV-1 Gag is associated with DRM and that disruption of Gag-raft interactions impairs HIV-1 particle production. However, the determinants of Gag-raft association remain undefined. In this study, we developed a novel epitope-based Gag multimerization assay to examine whether Gag assembly is essential for its association with lipid rafts. We observed that membrane-associated, full-length Gag is poorly detected by immunoprecipitation relative to non-membrane-bound Gag. This poor detection is due to assembly-driven masking of Gag epitopes, as denaturation greatly improves immunoprecipitation. Gag mutants lacking the Gag-Gag interaction domain located in the N terminus of the nucleocapsid (NC) were efficiently immunoprecipitated without denaturation, indicating that the epitope masking is caused by higher order Gag multimerization. We used this assay to examine the relationship between Gag assembly and Gag binding to total cellular membrane and DRM. Importantly, a multimerization-defective NC mutant displayed wild-type levels of membrane binding and DRM association, indicating that NC-mediated Gag multimerization is dispensable for association of Gag with membrane or DRM. We also demonstrate that different properties of sucrose and iodixanol membrane flotation gradients may explain some discrepancies regarding Gag-raft interactions. This report offers new insights into the association of HIV-1 Gag with membrane and with lipid rafts. PMID- 16254351 TI - Immunogenicity of recombinant fiber-chimeric adenovirus serotype 35 vector-based vaccines in mice and rhesus monkeys. AB - Preexisting immunity to adenovirus serotype 5 (Ad5) has been shown to suppress the immunogenicity of recombinant Ad5 (rAd5) vector-based vaccines for human immunodeficiency virus type 1 (HIV-1) in both preclinical studies and clinical trials. A potential solution to this problem is to utilize rAd vectors derived from rare Ad serotypes, such as Ad35. However, rAd35 vectors have appeared less immunogenic than rAd5 vectors in preclinical studies to date. In this study, we explore the hypothesis that the differences in immunogenicity between rAd5 and rAd35 vectors may be due in part to differences between the fiber proteins of these viruses. We constructed capsid chimeric rAd35 vectors containing the Ad5 fiber knob (rAd35k5) and compared the immunogenicities of rAd5, rAd35k5, and rAd35 vectors expressing simian immunodeficiency virus Gag and HIV-1 Env in mice and rhesus monkeys. In vitro studies demonstrated that rAd35k5 vectors utilized the Ad5 receptor CAR rather than the Ad35 receptor CD46. In vivo studies showed that rAd35k5 vectors were more immunogenic than rAd35 vectors in both mice and rhesus monkeys. These data suggest that the Ad5 fiber knob contributes substantially to the immunogenicity of rAd vectors. Moreover, these studies demonstrate that capsid chimeric rAd vectors can be constructed to combine beneficial immunologic and serologic properties of different Ad serotypes. PMID- 16254350 TI - Alpha/beta interferons regulate murine gammaherpesvirus latent gene expression and reactivation from latency. AB - Alpha/beta interferon (IFN-alpha/beta) protects the host from virus infection by inhibition of lytic virus replication in infected cells and modulation of the antiviral cell-mediated immune response. To determine whether IFN-alpha/beta also modulates the virus-host interaction during latent virus infection, we infected mice lacking the IFN-alpha/beta receptor (IFN-alpha/betaR(-/-)) and wild-type (wt; 129S2/SvPas) mice with murine gammaherpesvirus 68 (gammaHV68), a lymphotropic gamma-2-herpesvirus that establishes latent infection in B cells, macrophages, and dendritic cells. IFN-alpha/betaR(-/-) mice cleared low-dose intranasal gammaHV68 infection with wt kinetics and harbored essentially wt frequencies of latently infected cells in both peritoneum and spleen by 28 days postinfection. However, latent virus in peritoneal cells and splenocytes from IFN alpha/betaR(-/-) mice reactivated ex vivo with >40-fold- and 5-fold-enhanced efficiency, respectively, compared to wt cells. Depletion of IFN-alpha/beta from wt mice during viral latency also significantly increased viral reactivation, demonstrating an antiviral function of IFN-alpha/beta during latency. Viral reactivation efficiency was temporally regulated in both wt and IFN-alpha/betaR( /-) mice. The mechanism of IFN-alpha/betaR action was distinct from that of IFN gammaR, since IFN-alpha/betaR(-/-) mice did not display persistent virus replication in vivo. Analysis of viral latent gene expression in vivo demonstrated specific upregulation of the latency-associated gene M2, which is required for efficient reactivation from latency, in IFN-alpha/betaR(-/-) splenocytes. These data demonstrate that an IFN-alpha/beta-induced pathway regulates gammaHV68 gene expression patterns during latent viral infection in vivo and that IFN-alpha/beta plays a critical role in inhibiting viral reactivation during latency. PMID- 16254352 TI - Phenotypic, functional, and kinetic parameters associated with apparent T-cell control of human immunodeficiency virus replication in individuals with and without antiretroviral treatment. AB - The human immunodeficiency virus (HIV)-mediated immune response may be beneficial or harmful, depending on the balance between expansion of HIV-specific T cells and the level of generalized immune activation. The current study utilizes multicolor cytokine flow cytometry to study HIV-specific T cells and T-cell activation in 179 chronically infected individuals at various stages of HIV disease, including those with low-level viremia in the absence of therapy ("controllers"), low-level drug-resistant viremia in the presence of therapy (partial controllers on antiretroviral therapy [PCAT]), and high-level viremia ("noncontrollers"). Compared to noncontrollers, controllers exhibited higher frequencies of HIV-specific interleukin-2-positive gamma interferon-positive (IL 2(+) IFN-gamma(+)) CD4(+) T cells. The presence of HIV-specific CD4(+) IL-2(+) T cells was associated with low levels of proliferating T cells within the less differentiated T-cell subpopulations (defined by CD45RA, CCR7, CD27, and CD28). Despite prior history of progressive disease, PCAT patients exhibited many immunologic characteristics seen in controllers, including high frequencies of IL 2(+) IFN-gamma(+) CD4(+) T cells. Measures of immune activation were lower in all CD8(+) T-cell subsets in controllers and PCAT compared to noncontrollers. Thus, control of HIV replication is associated with high levels of HIV-specific IL-2(+) and IFN-gamma(+) CD4(+) T cells and low levels of T-cell activation. This immunologic state is one where the host responds to HIV by expanding but not exhausting HIV-specific T cells while maintaining a relatively quiescent immune system. Despite a history of advanced HIV disease, a subset of individuals with multidrug-resistant HIV exhibit an immunologic profile comparable to that of controllers, suggesting that functional immunity can be reconstituted with partially suppressive highly active antiretroviral therapy. PMID- 16254353 TI - Human immunodeficiency virus type 1 can establish latent infection in resting CD4+ T cells in the absence of activating stimuli. AB - Resting CD4(+) T cells are the best-defined reservoir of latent human immunodeficiency virus type 1 (HIV-1) infection, but how the reservoir is formed is unclear. Understanding how the reservoir of latently infected cells forms is critical because it is a major barrier to curing HIV infection. The system described here may provide an in vitro model of latent HIV-1 infection in resting CD4(+) T cells. We demonstrated that HIV-1 integrates into the genomes of in vitro-inoculated resting CD4(+) T cells that have not received activating stimuli and have not entered cell cycle stage G(1b). A percentage of the resting CD4(+) T cells that contain integrated DNA produce virus upon stimulation, i.e., are latently infected. Our results show that latent HIV-1 infection occurs in unstimulated resting CD4(+) T cells and suggest a new route for HIV-1 reservoir formation. PMID- 16254355 TI - Characterization of Parvovirus B19 genotype 2 in KU812Ep6 cells. AB - An infectious parvovirus B19 (B19V) genotype 2 variant was identified as a high titer contaminant in a human plasma donation. Genome analysis revealed a 138-bp insertion within the p6 promoter. The inserted sequence was represented by an additional 30 bp from the end of the inverted terminal repeat adjacent to a 108 bp element found also, in inverted orientation, at the extreme right end of the unique sequence of the genome. However, despite the profound variations in the promoter region, the pattern of gene expression and DNA replication did not differ between genotype 1 and genotype 2 in permissive erythroid KU812Ep6 cells. Capsid proteins of both genotypes differ in their amino acid sequences. However, equivalent kinetics of virus inactivation at 56 degrees C or pH 4 indicated a comparable physicochemical stability of virus capsids. Sera from six individuals infected by B19V genotype 1 were investigated on cross-neutralization of B19V genotype 2 in vitro. Similar neutralization of both B19V genotypes was observed in sera from three individuals, while the sera from three other individuals showed weaker cross-neutralization for genotype 2. In conclusion, the in vitro replication characteristics and physical stability of B19V capsids are very similar between human parvovirus B19 genotypes 1 and 2, and cross-neutralization indicates a close antigenic relation of genotypes 1 and 2. PMID- 16254354 TI - Protective cytotoxic T-cell responses induced by venezuelan equine encephalitis virus replicons expressing Ebola virus proteins. AB - Infection with Ebola virus causes a severe disease accompanied by high mortality rates, and there are no licensed vaccines or therapies available for human use. Filovirus vaccine research efforts still need to determine the roles of humoral and cell-mediated immune responses in protection from Ebola virus infection. Previous studies indicated that exposure to Ebola virus proteins expressed from packaged Venezuelan equine encephalitis virus replicons elicited protective immunity in mice and that antibody-mediated protection could only be demonstrated after vaccination against the glycoprotein. In this study, the murine CD8(+) T cell responses to six Ebola virus proteins were examined. CD8(+) T cells specific for Ebola virus glycoprotein, nucleoprotein, and viral proteins (VP24, VP30, VP35, and VP40) were identified by intracellular cytokine assays using splenocytes from vaccinated mice. The cells were expanded by restimulation with peptides and demonstrated cytolytic activity. Adoptive transfer of the CD8(+) cytotoxic T cells protected filovirus naive mice from challenge with Ebola virus. These data support a role for CD8(+) cytotoxic T cells as part of a protective mechanism induced by vaccination against six Ebola virus proteins and provide additional evidence that cytotoxic T-cell responses can contribute to protection from filovirus infections. PMID- 16254356 TI - Kaposi's sarcoma-associated herpesvirus K8beta is derived from a spliced intermediate of K8 pre-mRNA and antagonizes K8alpha (K-bZIP) to induce p21 and p53 and blocks K8alpha-CDK2 interaction. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) is a lymphotropic DNA tumor virus that induces Kaposi's sarcoma and AIDS-related primary effusion lymphoma. KSHV open reading frame 50 and K8 genes in early viral lytic infection express, respectively, a tricistronic and a bicistronic pre-mRNA, which undergo alternative splicing to create two major spliced mRNA isoforms, alpha and beta, by inclusion (beta) or exclusion (alpha) of an intron at nucleotides 75563 to 75645. This intron contains some suboptimal features, which cause the intron 5' splice site (ss) to interact weakly with U1 snRNA and the 3' ss to bind a U2 auxiliary factor, U2AF, with low affinity. Optimization of this intron in K8 (K8 intron 2) promoted the interaction of the 5' ss with U1 and the 3' ss with U2AF, resulting in a substantial increase in intron splicing. Splicing of K8 intron 2 has also been shown to be stimulated by the splicing of a downstream intron. This was confirmed by the insertion of a human beta-globin intron into the K8beta exon 3-exon 4 splice junction, which promoted splicing of K8beta intron 2 and conversion of the K8beta mRNA to the K8alpha mRNA that encodes a K-bZIP protein. Intron 2 contains a premature termination codon, yet the K8beta mRNA is insensitive to nonsense-mediated mRNA decay, suggesting that the truncated K8beta protein may have a biological function. Indeed, although the truncated K8beta protein is missing only a C-terminal leucine zipper domain from the K-bZIP, its expression antagonizes the ability of the K-bZIP to induce p53 and p21 and blocks K-bZIP-CDK2 interaction through interfering K8alpha mRNA production. PMID- 16254357 TI - Interaction between Brome mosaic virus proteins and RNAs: effects on RNA replication, protein expression, and RNA stability. AB - Brome mosaic virus (BMV) RNA replication has been examined in a number of systems, including Saccharomyces cerevisiae. We developed an efficient T-DNA based gene delivery system using Agrobacterium tumefaciens to transiently express BMV RNAs in Nicotiana benthamiana. The expressed RNAs can systemically infect plants and provide material to extract BMV replicase that can perform template dependent RNA-dependent RNA synthesis in vitro. We also expressed the four BMV encoded proteins from nonreplicating RNAs and analyzed their effects on BMV RNA accumulation. The capsid protein that coinfiltrated with constructs expressing RNA1 and RNA2 suppressed minus-strand levels but increased plus-strand RNA accumulation. The replication proteins 1a and 2a could function in trans to replicate and transcribe the BMV RNAs. None of the BMV proteins or RNA could efficiently suppress posttranscriptional silencing. However, 1a expressed in trans will suppress the production of a recombinant green fluorescent protein expressed from the nontranslated portions of BMV RNA1 and RNA2, suggesting that 1a may regulate translation from BMV RNAs. BMV replicase proteins 1a did not affect the accumulation of the BMV RNAs in the absence of RNA replication, unlike the situation reported for S. cerevisiae. This work demonstrates that the Agrobacterium-mediated gene delivery system can be used to study the cis- and trans-acting requirements for BMV RNA replication in plants and that significant differences can exist for BMV RNA replication in different hosts. PMID- 16254358 TI - Mutation of a single conserved nucleotide between the cloverleaf and internal ribosome entry site attenuates poliovirus neurovirulence. AB - The chemical synthesis of poliovirus (PV) cDNA combined with the cell-free synthesis of infectious particles yielded virus whose mouse neurovirulence was highly attenuated (J. Cello, A. V. Paul, and E. Wimmer, Science 297:1016-1018, 2002). Compared to the wild-type PV1 (Mahoney) [PV1(M)] sequence, the synthetic virus genome harbored 27 nucleotide (nt) changes deliberately introduced as genetic markers. Of the 27 nucleotide substitutions, the UA-to-GG exchanges at nucleotides 102/103, mapping to a region between the cloverleaf and the internal ribosome entry site (IRES) in the 5'-nontranslated region, were found to be involved in the observed attenuation phenotype in mice. The UA/GG mutation at nt 102/103 in the synthetic PV1(M) [sPV1(M)] background conferred also a ts phenotype of replication to the virus in human neuroblastoma cells. Conversely, the exchange of GG to wild-type (wt) UA at 102/103 in an sPV1(M) background restored wt neurovirulence in CD155 transgenic (tg) mice and suppressed the ts phenotype in SK-N-MC cells. All poliovirus variants replicated well in HeLa cells at the two temperatures, regardless of the sequence at the 102/103 locus. Analyses of variants isolated from sPV(M)-infected CD155 tg mice revealed that the G(102)G(103)-to-G(102)A(103) reversion alone reestablished the neurovirulent phenotype. This suggests that a single mutation is responsible for the observed change of the neurovirulence phenotype. sPV1(M) RNA is translated in cell extracts of SK-N-MC cells with significantly lower efficiency than PV1(M) RNA or sPV1(M) RNA with a G(102)-to-A(102) reversion. These studies suggest a function for the conserved nucleotide (A(103)) located between the cloverleaf and the IRES which is important for replication of PV in the central nervous system of CD155 tg mice and in human cells of neuronal origin. PMID- 16254359 TI - Predicted inactivation of viruses of relevance to biodefense by solar radiation. AB - UV radiation from the sun is the primary germicide in the environment. The goal of this study was to estimate inactivation of viruses by solar exposure. We reviewed published reports on 254-nm UV inactivation and tabulated the sensitivities of a wide variety of viruses, including those with double-stranded DNA, single-stranded DNA, double-stranded RNA, or single-stranded RNA genomes. We calculated D(37) values (fluence producing on average one lethal hit per virion and reducing viable virus to 37%) from all available data. We defined "size normalized sensitivity" (SnS) by multiplying UV(254) sensitivities (D(37) values) by the genome size, and SnS values were relatively constant for viruses with similar genetic composition. In addition, SnS values were similar for complete virions and their defective particles, even when the corresponding D(37) values were significantly different. We used SnS to estimate the UV(254) sensitivities of viruses for which the genome composition and size were known but no UV inactivation data were available, including smallpox virus, Ebola, Marburg, Crimean-Congo, Junin, and other hemorrhagic viruses, and Venezuelan equine encephalitis and other encephalitis viruses. We compiled available data on virus inactivation as a function of wavelength and calculated a composite action spectrum that allowed extrapolation from the 254-nm data to solar UV. We combined our estimates of virus sensitivity with solar measurements at different geographical locations to predict virus inactivation. Our predictions agreed with the available experimental data. This work should be a useful step to understanding and eventually predicting the survival of viruses after their release in the environment. PMID- 16254360 TI - Effect of alternating passage on adaptation of sindbis virus to vertebrate and invertebrate cells. AB - Mosquito-borne alphaviruses, which replicate alternately and obligately in mosquitoes and vertebrates, appear to experience lower rates of evolution than do many RNA viruses that replicate solely in vertebrates. This genetic stability is hypothesized to result from the alternating host cycle, which constrains evolution by imposing compromise fitness solutions in each host. To test this hypothesis, Sindbis virus was passaged serially, either in one cell type to eliminate host alteration or alternately between vertebrate (BHK) and mosquito (C6/36) cells. Following 20 to 50 serial passages, mutations were identified and changes in fitness were assessed using competition assays against genetically marked, surrogate parent viruses. Specialized viruses passaged in a single cell exhibited more mutations and amino acid changes per passage than those passaged alternately. Single host-adapted viruses exhibited fitness gains in the cells in which they specialized but fitness losses in the bypassed cell type. Most but not all viruses passaged alternately experienced lesser fitness gains than specialized viruses, with fewer mutations per passage. Clonal populations derived from alternately passaged viruses also exhibited adaptation to both cell lines, indicating that polymorphic populations are not required for simultaneous fitness gains in vertebrate and mosquito cells. Nearly all passaged viruses acquired Arg or Lys substitutions in the E2 envelope glycoprotein, but enhanced binding was only detected for BHK cells. These results support the hypothesis that arbovirus evolution may be constrained by alternating host transmission cycles, but they indicate a surprising ability for simultaneous adaptation to highly divergent cell types by combinations of mutations in single genomes. PMID- 16254361 TI - Noncytopathogenic pestivirus strains generated by nonhomologous RNA recombination: alterations in the NS4A/NS4B coding region. AB - Several studies have demonstrated that cytopathogenic (cp) pestivirus strains evolve from noncytopathogenic (noncp) viruses by nonhomologous RNA recombination. In addition, two recent reports showed the rapid emergence of noncp Bovine viral diarrhea virus (BVDV) after a few cell culture passages of cp BVDV strains by homologous recombination between identical duplicated viral sequences. To allow the identification of recombination sites from noncp BVDV strains that evolve from cp viruses, we constructed the cp BVDV strains CP442 and CP552. Both harbor duplicated viral sequences of different origin flanking the cellular insertion Nedd8*; the latter is a prerequisite for their cytopathogenicity. In contrast to the previous studies, isolation of noncp strains was possible only after extensive cell culture passages of CP442 and CP552. Sequence analysis of 15 isolated noncp BVDVs confirmed that all recombinant strains lack at least most of Nedd8*. Interestingly, only one strain resulted from homologous recombination while the other 14 strains were generated by nonhomologous recombination. Accordingly, our data suggest that the extent of sequence identity between participating sequences influences both frequency and mode (homologous versus nonhomologous) of RNA recombination in pestiviruses. Further analyses of the noncp recombinant strains revealed that a duplication of 14 codons in the BVDV nonstructural protein 4B (NS4B) gene does not interfere with efficient viral replication. Moreover, an insertion of viral sequences between the NS4A and NS4B genes was well tolerated. These findings thus led to the identification of two genomic loci which appear to be suited for the insertion of heterologous sequences into the genomes of pestiviruses and related viruses. PMID- 16254362 TI - Effect of macromolecular crowding agents on human immunodeficiency virus type 1 capsid protein assembly in vitro. AB - Previous studies on the self-assembly of capsid protein CA of human immunodeficiency virus type 1 (HIV-1) in vitro have provided important insights on the structure and assembly of the mature HIV-1 capsid. However, CA polymerization in vitro was previously observed to occur only at very high ionic strength. Here, we have analyzed the effects on CA assembly in vitro of adding unrelated, inert macromolecules (crowding agents), aimed at mimicking the crowded (very high macromolecular effective concentration) environment within the HIV-1 virion. Crowding agents induced fast and efficient polymerization of CA even at low (close to physiological) ionic strength. The hollow cylinders thus assembled were indistinguishable in shape and dimensions from those formed in dilute protein solutions at high ionic strength. However, two important differences were noted: (i) disassembly by dilution of the capsid-like particles was undetectable at very high ionic strength, but occurred rapidly at low ionic strength in the presence of a crowding agent, and (ii) a variant CA from a presumed infectious HIV-1 with mutations at the CA dimerization interface was unable to assemble at any ionic strength in the absence of a crowding agent; in contrast, this mutation allowed efficient assembly, even at low ionic strength, when a crowding agent was used. The use of a low ionic strength and inert macromolecules to mimic the crowded environment inside the HIV-1 virion may lead to a better in vitro evaluation of the effects of conditions, mutations or/and other molecules, including potential antiviral compounds, on HIV-1 capsid assembly, stability and disassembly. PMID- 16254363 TI - Posttranslational modification of alpha-dystroglycan, the cellular receptor for arenaviruses, by the glycosyltransferase LARGE is critical for virus binding. AB - The receptor for lymphocytic choriomeningitis virus (LCMV), the human pathogenic Lassa fever virus (LFV), and clade C New World arenaviruses is alpha-dystroglycan (alpha-DG), a cell surface receptor for proteins of the extracellular matrix (ECM). Specific posttranslational modification of alpha-DG by the glycosyltransferase LARGE is critical for its function as an ECM receptor. In the present study, we show that LARGE-dependent modification is also crucial for alpha-DG's function as a cellular receptor for arenaviruses. Virus binding involves the mucin-type domain of alpha-DG and depends on modification by LARGE. A crucial role of the LARGE-dependent glycosylation of alpha-DG for virus binding is found for several isolates of LCMV, LFV, and the arenaviruses Mobala and Oliveros. Since the posttranslational modification by LARGE is crucial for alpha DG recognition by both arenaviruses and the host-derived ligand laminin, it also influences competition between virus and laminin for alpha-DG. Hence, LARGE dependent glycosylation of alpha-DG has important implications for the virus-host cell interaction and the pathogenesis of LFV in humans. PMID- 16254364 TI - O Mannosylation of alpha-dystroglycan is essential for lymphocytic choriomeningitis virus receptor function. AB - Alpha-dystroglycan (alpha-DG) was identified as a common receptor for lymphocytic choriomeningitis virus (LCMV) and several other arenaviruses including the human pathogenic Lassa fever virus. Initial work postulated that interactions between arenavirus glycoproteins and alpha-DG are based on protein-protein interactions. We found, however, that susceptibility toward LCMV infection differed in various cell lines despite them expressing comparable levels of DG, suggesting that posttranslational modifications of alpha-DG would be involved in viral receptor function. Here, we demonstrate that glycosylation of alpha-DG, and in particular, O mannosylation, which is a rare type of O-linked glycosylation in mammals, is essential for LCMV receptor function. Cells that are defective in components of the O-mannosylation pathway showed strikingly reduced LCMV infectibility. As defective O mannosylation is associated with severe clinical symptoms in mammals such as congenital muscular dystrophies, it is likely that LCMV and potentially other arenaviruses may have selected this conserved and crucial posttranslational modification as the primary target structure for cell entry and infection. PMID- 16254365 TI - The human DEK proto-oncogene is a senescence inhibitor and an upregulated target of high-risk human papillomavirus E7. AB - The human DEK proto-oncogene is a nucleic acid binding protein with suspected roles in human carcinogenesis, autoimmune disease, and viral infection. Intracellular DEK functions, however, are poorly understood. In papillomavirus positive cervical cancer cells, downregulation of viral E6/E7 oncogene expression results in cellular senescence. We report here the specific repression of DEK message and protein levels in senescing human papillomavirus type 16- (HPV16-) and HPV18-positive cancer cell lines as well as in primary cells undergoing replicative senescence. Cervical cancer cell senescence was partially overcome by DEK overexpression, and DEK overexpression was sufficient for extending the life span of primary keratinocytes, supporting critical roles for this molecule as a senescence regulator. In order to determine whether DEK is a bona fide HPV oncogene target in primary cells, DEK expression was monitored in human keratinocytes transduced with HPV E6 and/or E7. The results identify high-risk HPV E7 as a positive DEK regulator, an activity that is not shared by low-risk HPV E7 protein. Experiments in mouse embryo fibroblasts recapitulated the observed E7-mediated DEK induction and demonstrated that both basal and E7 induced regulation of DEK expression are controlled by the retinoblastoma protein family. Taken together, our results suggest that DEK upregulation may be a common event in human carcinogenesis and may reflect its senescence inhibitory function. PMID- 16254366 TI - Human beta-defensins suppress human immunodeficiency virus infection: potential role in mucosal protection. AB - Beta-defensins are small (3 to 5 kDa in size) secreted antimicrobial and antiviral proteins that are components of innate immunity. Beta-defensins are secreted by epithelial cells, and they are expressed at high levels in several mucosae, including the mouth, where the concentration of these proteins can reach 100 microg/ml. Because of these properties, we wondered whether they could be part of the defenses that lower oral transmission of human immunodeficiency virus (HIV) compared to other mucosal sites. Our data show that select beta-defensins, especially human beta-defensin 2 (hBD2) and hBD3, inhibit R5 and X4 HIV infection in a dose-dependent manner at doses that are compatible with or below those measured in the oral cavity. We observed that beta-defensin treatment inhibited accumulation of early products of reverse transcription, as detected by PCR. We could not, however, detect any reproducible inhibition of env-mediated fusion, and we did not observe any modulation of HIV coreceptors following treatment with hBD1 and hBD2, in both resting and phytohemagglutinin-activated cells. Our data instead suggest that, besides a direct inactivation of HIV virions, hBD2 inhibits HIV replication in the intracellular environment. Therefore, we speculate that beta-defensins mediate a novel antiretroviral mechanism that contributes to prevention of oral HIV transmission in the oral cavity. Immunohistochemical data on hBD2 expression in oral mucosal tissue shows that hBD2 is constitutively expressed, forming a barrier layer across the epithelium in healthy subjects, while in HIV-positive subjects levels of hBD2 expression are dramatically diminished. This may predispose HIV-positive subjects to increased incidence of oral complications associated with HIV infection. PMID- 16254368 TI - PrPTSE distribution in a primate model of variant, sporadic, and iatrogenic Creutzfeldt-Jakob disease. AB - Human prion diseases, such as Creutzfeldt-Jakob disease (CJD), are neurodegenerative and fatal. Sporadic CJD (sCJD) can be transmitted between humans through medical procedures involving highly infected organs, such as the central nervous system. However, in variant CJD (vCJD), which is due to human contamination with the bovine spongiform encephalopathy (BSE) agent, lymphoreticular tissue also harbors the transmissible spongiform encephalopathy associated prion protein (PrP(TSE)), which poses a particularly acute risk for iatrogenic transmission. Two blood transfusion-related cases are already documented. In addition, the recent observation of PrP(TSE) in spleen and muscle in sCJD raised the possibility that peripheral PrP(TSE) is not limited to vCJD cases. We aimed to clarify the peripheral pathogenesis of human TSEs by using a nonhuman primate model which mimics human diseases. A highly sensitive enzyme linked immunosorbent assay was adapted to the detection of extraneural PrP(TSE). We show that affected organs can be divided into two groups. The first is peripheral organs accumulating large amounts of PrP(TSE), which represent a high risk of iatrogenic transmission. This category comprises only lymphoreticular organs in the vCJD/BSE model. The second is organs with small amounts of PrP(TSE) associated with nervous structures. These are the muscles, adrenal glands, and enteric nervous system in the sporadic, iatrogenic, and variant CJD models. In contrast to the first set of organs, this low level of tissue contamination is not strain restricted and seems to be linked to secondary centrifugal spread of the agent through nerves. It might represent a risk for iatrogenic transmission, formerly underestimated despite previous reports of low rates of transmission from peripheral organs of humans to nonhuman primates (5, 10). This study provides an additional experimental basis for the classification of human organs into different risk categories and a rational re-evaluation of current risk management measures. PMID- 16254367 TI - Rational attenuation of a morbillivirus by modulating the activity of the RNA dependent RNA polymerase. AB - Negative-strand RNA viruses encode a single RNA-dependent RNA polymerase (RdRp) which transcribes and replicates the genome. The open reading frame encoding the RdRp from a virulent wild-type strain of rinderpest virus (RPV) was inserted into an expression plasmid. Sequences encoding enhanced green fluorescent protein (EGFP) were inserted into a variable hinge of the RdRp. The resulting polymerase was autofluorescent, and its activity in the replication/transcription of a synthetic minigenome was reduced. We investigated the potential of using this approach to rationally attenuate a virus by inserting the DNA sequences encoding the modified RdRp into a full-length anti-genome plasmid from which a virulent virus (rRPV(KO)) can be rescued. A recombinant virus, rRPV(KO)L-RRegfpR, which grew at an indistinguishable rate and to an identical titer as rRPV(KO) in vitro, was rescued. Fluorescently tagged polymerase was visible in large cytoplasmic inclusions and beneath the cell membrane. Subcutaneous injection of 10(4) TCID(50) of the rRPV(KO) parental recombinant virus into cattle leads to severe disease symptoms (leukopenia/diarrhea and pyrexia) and death by 9 days postinfection. Animals infected with rRPV(KO)L-RRegfpR exhibited transient leukopenia and mild pyrexia, and the only noticeable clinical signs were moderate reddening of one eye and a slight ocular-nasal discharge. Viruses that expressed the modified polymerase were isolated from peripheral blood lymphocytes and eye swabs. This demonstrates that a virulent morbillivirus can be attenuated in a single step solely by modulating RdRp activity and that there is not necessarily a correlation between virus growth in vitro and in vivo. PMID- 16254369 TI - Long untranslated regions of the measles virus M and F genes control virus replication and cytopathogenicity. AB - Measles is still a major cause of mortality mainly in developing countries. The causative agent, measles virus (MeV), is an enveloped virus having a nonsegmented negative-sense RNA genome, and belongs to the genus Morbillivirus of the family Paramyxoviridae. One feature of the moribillivirus genomes is that the M and F genes have long untranslated regions (UTRs). The M and F mRNAs of MeV have 426 nucleotide-long 3' and 583-nucleotide-long 5' UTRs, respectively. Though these long UTRs occupy as much as approximately 6.4% of the virus genome, their function remains unknown. To elucidate the role of the long UTRs in the context of virus infection, we used the reverse genetics based on the virulent strain of MeV, and generated a series of recombinant viruses having alterations or deletions in the long UTRs. Our results showed that these long UTRs per se were not essential for MeV replication, but that they regulated MeV replication and cytopathogenicity by modulating the productions of the M and F proteins. The long 3' UTR of the M mRNA was shown to have the ability to increase the M protein production, promoting virus replication. On the other hand, the long 5' UTR of the F mRNA was found to possess the capacity to decrease the F protein production, inhibiting virus replication and yet greatly reducing cytopathogenicity. We speculate that the reduction in cytopathogenicity may be advantageous for MeV fitness and survival in nature. PMID- 16254370 TI - The Toll-like receptor 7 (TLR7) agonist, imiquimod, and the TLR9 agonist, CpG ODN, induce antiviral cytokines and chemokines but do not prevent vaginal transmission of simian immunodeficiency virus when applied intravaginally to rhesus macaques. AB - The initial host response to viral infection occurs after Toll-like receptors (TLRs) on dendritic cells (DC) are stimulated by viral nucleic acids (double stranded RNA, single-stranded RNA) and alpha interferon (IFN-alpha) and IFN-beta are produced. We hypothesized that pharmacologic induction of innate antiviral responses in the cervicovaginal mucosa by topical application of TLR agonists prior to viral exposure could prevent or blunt vaginal transmission of simian immunodeficiency virus (SIV). To test this hypothesis, we treated rhesus monkeys intravaginally with either the TLR9 agonist, CpG oligodeoxynucleotides (ODN), or the TLR7 agonist, imiquimod. Both immune modifiers rapidly induced IFN-alpha and other antiviral effector molecules in the cervicovaginal mucosa of treated animals. However, both CpG ODN and imiquimod also induced proinflammatory cytokine expression in the cervicovaginal mucosa. In the vaginal mucosa of imiquimod-treated monkeys, we documented a massive mononuclear cell infiltrate consisting of activated CD4(+) T cells, DC, and beta-chemokine-secreting cells. After vaginal SIV inoculation, all TLR agonist-treated animals became infected and had plasma vRNA levels that were higher than those of control monkeys. We conclude that induction of mucosal innate immunity including an IFN-alpha response is not sufficient to prevent sexual transmission of human immunodeficiency virus. PMID- 16254371 TI - Role of Notch signal transduction in Kaposi's sarcoma-associated herpesvirus gene expression. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) RTA transcription factor is recruited to its responsive elements through interaction with a Notch-mediated transcription factor, RBP-Jkappa, indicating that RTA mimics cellular Notch signal transduction to activate viral lytic gene expression. To test whether cellular Notch signal transduction and RTA are functionally exchangeable for viral gene expression, human Notch intracellular (hNIC) domain that constitutively activates RBP-Jkappa transcription factor activity was expressed in KSHV-infected primary effusion lymphoma BCBL1 cells (TRExBCBL1-hNIC) in a tetracycline-inducible manner. Gene expression profiling showed that like RTA, hNIC robustly induced expression of a number of viral genes, including viral interleukin 6 (vIL-6), K3, and K5. Unlike RTA, however, hNIC was not capable of evoking the full repertoire of lytic viral gene expression and thereby lytic replication. To further understand the role of Notch signal transduction in KSHV gene expression, vIL-6 growth factor and K5 immune modulator genes were selected for detailed analysis. Despite the presence of multiple RBP-Jkappa binding sites, hNIC targeted the specific RBP-Jkappa binding sites of vIL-6 and K5 promoter regions to regulate their gene expression. These results indicate that cellular Notch signal transduction not only is partially exchangeable with RTA in regard to activation of viral lytic gene expression but also provides a novel expression profile of KSHV growth and immune deregulatory genes that is likely different from that of RTA-independent standard latency program as well as RTA-dependent lytic reproduction program. PMID- 16254372 TI - Establishment and maintenance of Kaposi's sarcoma-associated herpesvirus latency in B cells. AB - Kaposi's sarcoma (KS)-associated herpesvirus (KSHV) is the infectious cause of Kaposi's sarcoma and is also associated with two B-cell lymphoproliferative diseases, primary effusion lymphoma and the plasmablastic form of multicentric Castleman's disease. KSHV is also found in the B-cell fraction of peripheral blood mononucleocytes of some KS patients. Despite in vivo infection of B cells and the ability of KSHV to infect many cell types in culture, to date B cells in culture have been resistant to KSHV infection. However, as shown here, the lack of infection is not due to the inability of B cells to support latent KSHV infection. When KSHV DNA is introduced into B cells, the virus is maintained as an episome and can establish and maintain latency over the course of months. As in all primary effusion lymphoma cell lines, there is a low level of spontaneous lytic replication in latently infected BJAB cells. Importantly, viral gene expression is similar to that of primary effusion lymphoma cell lines. Furthermore, the virus can be reactivated to higher levels with specific stimuli and transmitted to other cells, indicating that this is a productive infection. Thus B cells in culture are capable of establishing, maintaining, and reactivating from latency. These studies provide a controlled system to analyze how KSHV alters B cells during KSHV latency and reactivation. PMID- 16254373 TI - Genomic analysis of anti-hepatitis B virus (HBV) activity by small interfering RNA and lamivudine in stable HBV-producing cells. AB - Hepatitis B virus (HBV) causes acute and chronic hepatitis and hepatocellular carcinoma. Small interfering RNA (siRNA) and lamivudine have been shown to have anti-HBV effects through different mechanisms. However, assessment of the genome wide effects of siRNA and lamivudine on HBV-producing cell lines has not been reported, which may provide a clue to interrogate the HBV-cell interaction and to evaluate the siRNA's side effect as a potential drug. In the present study, we designed seven siRNAs based on the conserved HBV sequences and tested their effects on the expression of HBV genes following sorting of siRNA-positive cells. Among these seven siRNAs, siRNA-1 and siRNA-7 were found to effectively suppress HBV gene expression. We further addressed the global gene expression changes in stable HBV-producing cells induced by siRNA-1 and siRNA-7 by use of human genome wide oligonucleotide microarrays. Data from the gene expression profiling indicated that siRNA-1 and siRNA-7 altered the expression of 54 and 499 genes, respectively, in HepG2.2.15 cells, which revealed that different siRNAs had various patterns of gene expression profiles and suggested a complicated influence of siRNAs on host cells. We further observed that 18 of these genes were suppressed by both siRNA-1 and siRNA-7. Interestingly, seven of these genes were originally activated by HBV, which suggested that these seven genes might be involved in the HBV-host cell interaction. Finally, we have compared the effects of siRNA and lamivudine on HBV and host cells, which revealed that siRNA is more effective at inhibiting HBV expression at the mRNA and protein level in vitro, and the gene expression profile of HepG2.2.15 cells treated by lamivudine is totally different from that seen with siRNA. PMID- 16254374 TI - Novel type of hepatitis B virus mutation: replacement mutation involving a hepatocyte nuclear factor 1 binding site tandem repeat in chronic hepatitis B virus genotype E. AB - The genetic diversity of hepatitis B virus (HBV) strains has evolved through mutations such as point mutations, deletions or insertions, and recombination. We identified and characterized a novel type of mutation which is a complex of external insertion, deletion, and internal duplication in sequences from one of six patients with chronic hepatitis B virus genotype E (HBV/E). We provisionally named this mutation a "replacement mutation"; the core promoter upstream regulatory sequence/basic core promoter was replaced with a part of the S1 promoter covering the hepatocyte nuclear factor 1 (HNF1) binding site, followed by a tandem repeat of the HNF1 site. A longitudinal analysis of the HBV population over 6 years showed the clonal change from wild-type HBV/E to replacement-mutant type, resulting in a lower hepatitis B (HB) e antigen titer, a high HBV DNA level in serum, and progression of liver fibrosis. In an in vitro study using a replication model, the replacement-mutant HBV showed higher replication levels than the wild-type HBV/E replicon, probably mediated by altered transcription factor binding. Additionally, this HNF1 site replacement mutation was associated with excessive HB nucleocapsid protein expression in hepatocytes, in both in vivo and in vitro studies. This novel mutation may be specific to HBV genotype E, and its prevalence requires further investigation. PMID- 16254375 TI - Rabies virus P protein interacts with STAT1 and inhibits interferon signal transduction pathways. AB - Rabies virus P protein is a cofactor of RNA polymerase. We investigated other potential roles of P (CVS strain) by searching for cellular partners using two hybrid screening. We isolated a cDNA encoding the signal transducer and activator of transcription 1 (STAT1) that is a critical component of interferon type I (IFN alpha/beta) and type II (IFN-gamma) signaling. We confirmed this interaction by glutathione S-transferase-pull-down assay. Deletion mutant analysis indicated that the carboxy-terminal part of P interacted with a region containing the DNA binding domain and the coiled-coil domain of STAT1. The expression of P protein inhibits IFN-alpha- and IFN-gamma-induced transcriptional responses, thus impairing the IFN-induced antiviral state. Mechanistic studies indicate that P protein does not induce STAT1 degradation and does not interfere with STAT1 phosphorylation but prevents IFN-induced STAT1 nuclear accumulation. These results indicate that rabies P protein overcomes the antiviral response of the infected cells. PMID- 16254376 TI - Actin cytoskeleton is involved in targeting of a viral Hsp70 homolog to the cell periphery. AB - The cell-to-cell movement of plant viruses involves translocation of virus particles or nucleoproteins to and through the plasmodesmata (PDs). As we have shown previously, the movement of the Beet yellows virus requires the concerted action of five viral proteins including a homolog of cellular approximately 70 kDa heat shock proteins (Hsp70h). Hsp70h is an integral component of the virus particles and is also found in PDs of the infected cells. Here we investigate subcellular distribution of Hsp70h using transient expression of Hsp70h fused to three spectrally distinct fluorescent proteins. We found that fluorophore-tagged Hsp70h forms motile granules that are associated with actin microfilaments, but not with microtubules. In addition, immobile granules were observed at the cell periphery. A pairwise appearance of these granules at the opposite sides of cell walls and their colocalization with the movement protein of Tobacco mosaic virus indicated an association of Hsp70h with PDs. Treatment with various cytoskeleton specific drugs revealed that the intact actomyosin motility system is required for trafficking of Hsp70h in cytosol and its targeting to PDs. In contrast, none of the drugs interfered with the PD localization of Tobacco mosaic virus movement protein. Collectively, these findings suggest that Hsp70h is translocated and anchored to PDs in association with the actin cytoskeleton. PMID- 16254377 TI - CD46 is a cellular receptor for all species B adenoviruses except types 3 and 7. AB - The 51 human adenovirus serotypes are divided into six species (A to F). Adenovirus serotypes from all species except species B utilize the coxsackie adenovirus receptor for attachment to host cells in vitro. Species B adenoviruses primarily cause ocular and respiratory tract infections, but certain serotypes are also associated with renal disease. We have previously demonstrated that adenovirus type 11 (species B) uses CD46 (membrane cofactor protein) as a cellular receptor instead of the coxsackie-adenovirus receptor (A. Segerman et al., J. Virol. 77:9183-9191, 2003). In the present study, we found that transfection with human CD46 cDNA rendered poorly permissive Chinese hamster ovary cells more permissive to infection by all species B adenovirus serotypes except adenovirus types 3 and 7. Moreover, rabbit antiserum against human CD46 blocked or efficiently inhibited all species B serotypes except adenovirus types 3 and 7 from infecting human A549 cells. We also sequenced the gene encoding the fiber protein of adenovirus type 50 (species B) and compared it with the corresponding amino acid sequences from selected serotypes, including all other serotypes of species B. From the results obtained, we conclude that CD46 is a major cellular receptor on A549 cells for all species B adenoviruses except types 3 and 7. PMID- 16254378 TI - Intracellular interference of infectious bursal disease virus. AB - A search for dominant-negative mutant polypeptides hampering infectious bursal disease virus (IBDV) replication has been undertaken. We have found that expression of a mutant version of the VP3 structural polypeptide known as VP3/M3, partially lacking the domain responsible for the interaction with the virus encoded RNA polymerase, efficiently interferes with the IBDV replication cycle. Transformed cells stably expressing VP3/M3 show a significant reduction (up to 96%) in their ability to support IBDV growth. Our findings provide a new tool for the characterization of the IBDV replication cycle and might facilitate the generation of genetically modified chicken lines with a reduced susceptibility to IBDV infection. PMID- 16254379 TI - An N-linked glycoprotein with alpha(2,3)-linked sialic acid is a receptor for BK virus. AB - BK virus (BKV) is a common human polyomavirus infecting >80% of the population worldwide. Infection with BKV is asymptomatic, but reactivation in renal transplant recipients can lead to polyomavirus-associated nephropathy. In this report, we show that enzymatic removal of alpha(2,3)-linked sialic acid from cells inhibited BKV infection. Reconstitution of asialo cells with alpha(2,3) specific sialyltransferase restored susceptibility to infection. Inhibition of N linked glycosylation with tunicamycin reduced infection, but inhibition of O linked glycosylation did not. An O-linked-specific alpha(2,3)-sialyltransferase was unable to restore infection in asialo cells. Taken together, these data indicate that an N-linked glycoprotein containing alpha(2,3)-linked sialic acid is a critical component of the cellular receptor for BKV. PMID- 16254380 TI - Retroviral restriction factor TRIM5alpha is a trimer. AB - The retrovirus restriction factor TRIM5alpha targets the viral capsid soon after entry. Here we show that the TRIM5alpha protein oligomerizes into trimers. The TRIM5alpha coiled-coil and B30.2(SPRY) domains make important contributions to the formation and/or stability of the trimers. A functionally defective TRIM5alpha mutant with the RING and B-box 2 domains deleted can form heterotrimers with wild-type TRIM5alpha, accounting for the observed dominant negative activity of the mutant protein. Trimerization potentially allows TRIM5alpha to interact with threefold pseudosymmetrical structures on retroviral capsids. PMID- 16254381 TI - Murine coronavirus with an extended host range uses heparan sulfate as an entry receptor. AB - Only a relatively few mutations in its spike protein allow the murine coronavirus to switch from a murine-restricted tropism to an extended host range by being passaged in vitro. One such virus that we studied had acquired two putative heparan sulfate-binding sites while preserving another site in the furin-cleavage motif. The adaptation of the virus through the use of heparan sulfate as an attachment/entry receptor was demonstrated by increased heparin binding as well as by inhibition of infection through treatment of cells and the virus with heparinase and heparin, respectively. PMID- 16254382 TI - Transcripts encoding K12, v-FLIP, v-cyclin, and the microRNA cluster of Kaposi's sarcoma-associated herpesvirus originate from a common promoter. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) is the causative agent of three malignancies associated with AIDS and immunosuppression. Tumor cells harbor latent virus and express kaposin (open reading frame [ORF] K12), v-FLIP (ORF 71), v-Cyclin (ORF 72), and latency-associated nuclear antigen (LANA; ORF 73). ORFs 71 to 73 are transcribed as multicistronic RNAs initiating from adjacent constitutive and inducible promoters upstream of ORF 73. Here we characterize a third promoter embedded within the ORF 71-to-73 transcription unit specifying transcripts that encode ORF 71/72 or K12. These transcripts may also be the source of 11 microRNAs arranged as a cluster between K12 and ORF 71. Our studies reveal a complex arrangement of interlaced transcription units, incorporating four important protein-encoding genes required for latency and pathogenesis and the entire KSHV microRNA repertoire. PMID- 16254384 TI - Finding, evaluating, and using research for best practice. PMID- 16254383 TI - The herpes simplex virus type 1 locus that encodes the latency-associated transcript enhances the frequency of encephalitis in male BALB/c mice. AB - Herpes simplex virus type 1 (HSV-1) is the leading cause of virus-induced encephalitis; however, the viral genes that regulate encephalitis have not been well characterized. In this study, we tested whether the LAT (latency-associated transcript) locus regulates the frequency of encephalitis in male or female mice. Male BALB/c mice are more susceptible to HSV-1-induced encephalitis than age matched female BALB/c mice. Deletion of LAT coding sequences reduced the frequency of encephalitis. A recombinant virus containing the first 1.5 kb of the LAT coding sequence induces levels of encephalitis in male BALB/c mice similar to those induced by wild-type HSV-1. PMID- 16254385 TI - Obese ICU patients: resource utilization and outcomes. AB - This project described prospectively obese, critically ill patients and the resources critical care nurses used to care for these challenging patients. It also examined the relationship between resources used by nurses and patient outcomes, including complications and length of stay. Forty-three participants were enrolled. Patients with a body mass index (BMI) 40 kg/m2 used the majority of equipment and personnel resources and experienced a prolonged length of stay. The most common equipment used was a specialty bed or mattress; the most common complications were related to the pulmonary system. Initial use of multiple resources may indicate a patient at risk for adverse outcomes. Nurses can use findings to anticipate care needs and develop interventions, such as optimal positioning, to avoid adverse outcomes. PMID- 16254386 TI - Benefits of a school-based health center in a preschool. AB - Although school-based health centers (SBHCs) deliver health care to vulnerable children, their effectiveness has not been well documented. This study compared the benefits of an SBHC with a School Health Survey and selected HEDIS measures in preschool children with and without access to an SBHC. Preschoolers with access to an SBHC (N = 130) and preschoolers without access (N = 131) were compared on (a) HEDIS measures including well-child care, immunizations, dental care, and smoke exposure; (b) measures of access and use of physical and mental health services; (c) satisfaction with health care; (d) barriers and facilitators to care; and (e) health insurance. Significant differences were found in parents' perceptions of children's physical and emotional health, self-esteem, incidence of behavioral problems, difficulty in obtaining care, number of hospitalizations, and satisfaction with care received. Findings suggest that holistic services provided by an SBHC positively impact the health of vulnerable preschool children. PMID- 16254387 TI - Counteracting fragmentation in the care of people with moderate and severe dementia. AB - Symptoms such as amnesia, agnosia, apraxia, and aphasia may lead to a fragmented experience and actions among people with moderate and severe dementia. The aim of this study was to explore the interactions where fragmentation occurred and how caregivers counteract fragmentation. The observation notes from participant observations were analyzed using interpretive content analysis. Fragmentation was noted if the patients showed that they did not recognize what was going on, the people involved, the things used in the action, or did not recognize themselves in the situation. Care providers could counteract fragmentation by a caring based on attentive interest in the interaction, valuing the person behind the dementia disease, using an individual perspective considering the impact of the dementia disease, and striving for mutual interpretation of the shared situation. Caring based on these assumptions could help the patients to keep their world together. PMID- 16254388 TI - Actual interaction and client centeredness in home care. AB - The purpose of this study is to explore client-nurse interaction from a client perspective with respect to client-centered care. A grounded theory study was conducted with Dutch clients who were chronically ill and receiving home care. Data were collected by focus interviews with 8 client informants, participatory observations with 45 clients, and semistructured interviews with 6 clients. The core category actual interaction was identified. Six patterns of actual interaction were distinguished. Changes in actual interaction could be related to changes in desired participation by the client and in allowed client participation by the professional. From the client's perspective, client centeredness means congruence between desired and allowed participation. Congruence was experienced with consent, dialogue, and consuming. Congruence is not necessarily synonymous with promoting patient participation or with doing as the client wants. Ongoing attentiveness, responsiveness, promotion of client autonomy, and being a critical caregiver are recommended. PMID- 16254389 TI - An information theoretic approach to macromolecular modeling: I. Sequence alignments. AB - We are interested in applying the principles of information theory to structural biology calculations. In this article, we explore the information content of an important computational procedure: sequence alignment. Using a reference state developed from exhaustive sequences, we measure alignment statistics and evaluate gap penalties based on first-principle considerations and gap distributions. We show that there are different gap penalties for different alphabet sizes and that the gap penalties can depend on the length of the sequences being aligned. In a companion article, we examine the information content of molecular force fields. PMID- 16254390 TI - An information theoretic approach to macromolecular modeling: II. Force fields. AB - In this article, we explore the information content of molecular force-field calculations. We make use of exhaustive lattice models of molecular conformations and reduced alphabet sequences to determine the relative resolving power of pairwise interaction-based force fields. We find that sequence-specific interactions that operate over longer distances offer greater amounts of information than nearest-neighbor or non-sequence-specific interactions. In a companion article in this issue, we explored the information content of sequence alignment procedures and the calculation of gap penalties. Both articles have implications for protein and nucleic-acid computations. PMID- 16254391 TI - Insulin particle formation in supersaturated aqueous solutions of poly(ethylene glycol). AB - Protein microspheres are of particular utility in the field of drug delivery. A novel, completely aqueous, process of microsphere fabrication has been devised based on controlled phase separation of protein from water-soluble polymers such as polyethylene glycols. The fabrication process results in the formation of spherical microparticles with narrow particle size distributions. Cooling of preheated human insulin-poly(ethylene glycol)-water solutions results in the facile formation of insulin particles. To map out the supersaturation conditions conducive to particle nucleation and growth, we determined the temperature- and concentration-dependent boundaries of an equilibrium liquid-solid phase separation. The kinetics of formation of microspheres were followed by dynamic and continuous-angle static light scattering techniques. The presence of PEG at a pH that was close to the protein's isoelectric point resulted in rapid nucleation and growth. The time elapsed from the moment of creation of a supersaturated solution and the detection of a solid phase in the system (the induction period, t(ind)) ranged from tens to several hundreds of seconds. The dependence of t(ind) on supersaturation could be described within the framework of classical nucleation theory, with the time needed for the formation of a critical nucleus (size <10 nm) being much longer than the time of the onset of particle growth. The growth was limited by cluster diffusion kinetics. The interfacial energies of the insulin particles were determined to be 3.2-3.4 and 2.2 mJ/m(2) at equilibrium temperatures of 25 and 37 degrees C, respectively. The insulin particles formed as a result of the process were monodisperse and uniformly spherical, in clear distinction to previously reported processes of microcrystalline insulin particle formation. PMID- 16254392 TI - Imaging spinal cord damage in multiple sclerosis. AB - During the past 2 decades, the considerable improvement of magnetic resonance (MR) technology and the development of new MR strategies capable of providing an in vivo overall assessment of multiple sclerosis (MS) pathology have allowed us to obtain important novel pieces of information on disease evolution in the brain. However, despite this, the correlation between brain MR imaging metrics and clinical disability are still suboptimal. A reason for this discrepancy might be the involvement of clinically eloquent structures, such as the spinal cord, which owing to technical challenges have not been extensively studied using MR imaging until very recently. An objective and accurate estimate of the presence and extent of spinal cord damage might indeed contribute to increasing the strength of the correlations between clinical and MRI metrics. This review summarizes the main results obtained from the application of conventional and modern MR-based techniques for the evaluation of spinal cord damage in MS. PMID- 16254393 TI - Talairach-based parcellation of neonatal brain magnetic resonance imaging data: validation of a new approach. AB - BACKGROUND AND PURPOSE: Talairach-based parcellation (TP) of human brain magnetic resonance imaging (MRI) data has been used increasingly in clinical research to make regional measurements of brain structures in vivo. Recently, TP has been applied to pediatric research to elucidate the changes in regional brain volumes related to several neurological disorders. However, all freely available tools have been designed to parcellate adult brain MRI data. Parcellation of neonatal MRI data is very challenging owing to the lack of strong signal contrast, variability in signal intensity within tissues, and the small size and thus difficulty in identifying small structures used as landmarks for TP. Hence the authors designed and validated a new interactive tool to parcellate brain MRI data from newborns and young infants. METHODS: The authors' tool was developed as part of a postprocessing pipeline, which includes registration of multichannel MR images, segmentation, and parcellation of the segmented data. The tool employs user-friendly interactive software to visualize and assign the anatomic landmarks required for parcellation, after which the planes and parcels are generated automatically by the algorithm. The authors then performed 3 sets of validation experiments to test the precision and reliability of their tool. RESULTS: Validation experiments of intra-and interrater reliability on data obtained from newborn and 1-year-old children showed a very high sensitivity of >95% and specificity >99.9%. The authors also showed that rotating and reformatting the original MRI data results in a statistically significant difference in parcel volumes, demonstrating the importance of using a tool such as theirs that does not require realignment of the data prior to parcellation. CONCLUSIONS: To the authors' knowledge, the presented approach is the first TP method that has been developed and validated specifically for neonatal brain MRI data. Their approach would also be valuable for the analysis of brain MRI data from older children and adults. PMID- 16254394 TI - Emergent basilar artery and bilateral posterior cerebral artery angioplasty, urokinase thrombolysis, and stenting for acute basilar artery occlusion secondary to diagnostic cardiac catheterization: case presentation. AB - This case presentation describes the use of emergent neurointerventional procedures to revascularize a patient who developed a symptomatic stroke during coronary artery angiography. Rapid identification of a neurologic deficit in the coronary angiography suite and notification of a skilled neurointerventionalist permit for early revascularization and improved patient outcomes. PMID- 16254395 TI - Application of clinical scanners in rats: experimental carotid imaging using magnetic resonance imaging, spiral computed tomography, and color duplex ultrasound. AB - BACKGROUND AND PURPOSE: Noninvasive small animal imaging allows for reduction of the required numbers of animals in research by providing the possibility of long term follow-up at various time points. Additionally, correlation to the investigated respective human disease is possible as equivalent equipment is employed. The authors therefore evaluate feasibility and potential of color duplex sonography, computed tomography angiography (CTA), and magnetic resonance angiography (MRA) by the use of clinical scanners for carotid artery imaging in rats. METHODS: Male Wistar rats (n = 17) were subjected to color duplex sonography, CTA, and MRA of the common carotid artery (CCA) and the carotid bifurcation. Clinical scanners were used for the experiments and optimal parameter settings evaluated accounting for the different size of the animals. The applied imaging methods were analyzed in regard to image quality and practicability in laboratory settings. RESULTS: The CCA could be clearly displayed by all imaging modalities in all rats. Duplex sonography provided distinct images and reproducible basic functional information. CTA and MRA provided distinct images of the CCA and the carotid bifurcation in both axial and reconstructed 3-dimensional images. The authors further describe different indications for these imaging methods regarding spatial resolution, acquisition times, possible scanning range, and application of contrast agent. CONCLUSIONS: Color duplex sonography, CTA, and MRA are all feasible methods for imaging of the carotid arteries in rats. Images of sufficient clarity and resolution could be obtained by the use of clinical scanners, yielding information about vessel size, direction of blood flow, and adjacent structures. Further studies need to be performed that address investigations of pathological conditions such as flow disturbances or vessel stenosis. PMID- 16254396 TI - Transcranial sonographic monitoring of hemorrhagic transformation in patients with acute middle cerebral artery infarction. AB - BACKGROUND AND PURPOSE: Previous studies indicate the potential of transcranial sonography (TCS) to detect cerebrovascular disease. The authors conducted this patient study to evaluate the diagnostic potential of gray-scale TCS in depicting hemorrhagic transformation (HT) in the early phase of middle cerebral artery infarction. METHODS: TCS was performed in 32 patients with acute ischemic stroke in the middle cerebral artery territory less than 12, 24 +/- 4, 72 +/- 6, and 120 +/- 12 hours after symptom onset (SONOS 5500, S4 probe, 16 cm investigation depth). Hemorrhagic transformation was identified as hyperechogenicity in the MCA territory, and the echogenicity of these areas was assessed. In addition, the dislocation of the third ventricle (midline shift, MLS) was assessed by TCS. Size and localization of infarction were determined by cranial computed tomography (CCT). RESULTS: In 10 of 11 patients, TCS detected HT as confirmed by CCT. In 1 patient, TCS provided a false-positive result. In another patient, TCS was unable to detect the hemorrhage (sensitivity, 91%; specificity, 95%). The echointensity of HT increased over time. MLS measurement failed to predict fatal outcome in 1 patient. CONCLUSIONS: TCS is a promising tool for depicting HT in patients with acute hemispheric stroke and might be suitable for monitoring purposes. PMID- 16254397 TI - The prognostic significance of visible infarction on computed tomography following lacunar stroke: results of a long-term follow-up study. AB - BACKGROUND AND PURPOSE: Substantial variability in functional outcome and relatively few factors predictive of death or degree of recovery have been observed in patients with lacunar stroke. Such indicators are of great use in the selection of optimal rehabilitation strategies after stroke. Although computed tomography (CT) of patients with a clinical diagnosis of lacunar stroke performed within the first 10 days shows evidence of cerebral infarction in 50% to 60%, the prognostic significance of a visible ischemic lesion on CT is unclear. METHODS: 633 patients who presented with symptoms consistent with lacunar stroke between June 1990 and February 1998 were studied. One hundred fourteen patients imaged with magnetic resonance, 41 patients with nonischemic diagnoses (hemorrhage or tumor), 57 patients imaged within 12 hours of ictus, and 17 patients with incomplete follow-up were excluded from the analysis. The remaining 404 patients were divided into 2 groups, depending on the appearance of the CT scan. Patients with a low-attenuation area on the CT scan consistent with an ischemic lesion in an appropriate region of the brain to explain the presenting symptoms were classified as "CT positive." Patients with either a normal CT scan of the brain or a scan that showed a lesion in an area inconsistent with the presenting symptoms were classified as "CT negative." A series of known or suspected prognostic factors were recorded for each patient: blood pressure, age, smoking, plasma glucose level, serum cholesterol level, and serum triglyceride level. Delay from stroke onset to scanning was also noted. The authors considered 3 outcome measures: survival time, outcome at 6 months after the stroke, and total length of hospital stay for the stroke admission. Six-month outcome was categorized as good (alive at home) or poor (alive in care or dead). RESULTS: There was no difference in survival between the 2 groups (P= .29, log-rank test). After adjusting for other significant prognostic factors (age; relative hazard per additional decade 1.67, P< .0001: plasma glucose level; relative hazard per additional mmol/l 1.08, P= .03) in a proportional hazards model, presence of visible infarction remained nonsignificant (relative hazard 0.84, P= .40). After adjustment for the other significant factor (age, P= .0001), there was no significant difference in 6-month outcome between CT positive and CT negative patients (P= .61). Median total length of hospital stay was not significantly different between the 2 groups (CT positive, 9 days; CT negative, 8 days; Mann Whitney test, P= .29). CONCLUSION: The authors conclude that in their cohort of patients, having corrected for other prognostic variables, the presence of visible infarction on CT brain scan performed between 12 hours and 30 days of onset of lacunar symptoms is not predictive of duration of hospital stay or of longer term outcome. PMID- 16254398 TI - Transcranial Doppler ultrasonographic evaluation of middle cerebral artery hemodynamics during mild hypothermia. AB - BACKGROUND AND PURPOSE: Induced hypothermia holds promise as an effective neuroprotective strategy following cerebral ischemia. The effect of mild hypothermia on cerebral hemodynamics is not well known. The authors investigated the influence of brain temperature on middle cerebral artery (MCA) mean flow velocity (MCA FV) and pulsatility index (MCA PI) in nonintubated, healthy volunteers undergoing mild induced hypothermia. METHODS: Mild hypothermia (target tympanic membrane temperature [T tym] degrees C) was induced in subjects using the Arctic Sun Temperature Management System (Medivance, Inc, Louisville, CO). MCA FV and MCA PI were recorded bilaterally with a 2 MHz pulsed probe every 30 minutes via the transtemporal window. RESULTS: Eighteen subjects (8 males, 10 females) 32 +/- 8 years of age were studied. Multivariate analysis indicated that MCA FV increased with increasing change in temperature (baseline tympanic temperature-tympanic temperature [DeltaT tym]) (P< .001), heart rate (HR) (P< .001), end-tidal CO 2(P= .025), arterial oxygen saturation (O2%) (P= .001), and with decreasing mean arterial blood pressure (P= .004). Multivariate analysis also indicated that ln(MCA PI) (natural logarithm of MCA PI) decreased with decreasing T tym(P< .001) and increasing HR (P< .001). CONCLUSIONS: Mild induced hypothermia is associated with an increase in MCA FV and a decrease in MCA PI. The increase in MCA FV may be partially due to microcirculatory vasodilation. PMID- 16254399 TI - Diffusion-weighted imaging with calculated apparent diffusion coefficient of enhancing extra-axial masses. AB - BACKGROUND AND PURPOSE: Sometimes intracranial contrast-enhancing tumors like meningiomas, metastases, lymphomas, and schwannomas can mimic each other. It was the aim of the present study to investigate if intracranial contrast-enhancing lesions can be reliably differentiated with the help of diffusion-weighted imaging with calculated apparent diffusion coefficients (ADCs). METHODS: 29 patients (ages ranging from 22 to 82 years, mean age of 58.6 years) were included. Nine meningiomas, 7 metastases, 6 lymphomas, and 7 schwannomas were investigated. The ADC value in the lesions and in the perifocal edema was analyzed. RESULTS: For the lymphomas, the authors measured the lowest ADC values in the contrast-enhancing part (0.59 +/- 0.09 . 10(-3) mm2/sec). The meningiomas showed a mean ADC value of 0.98 +/- 0.18 . 10(-3) mm2/sec. The schwannomas and metastases showed higher ADC values of 1.33 +/- 0.28 . 10(-3) mm2/sec and 1.05 +/ 0.20 . 10(-3) mm2/sec. The authors saw a statistically significant difference between lymphomas, meningiomas, and metastases concerning the ADC values in the contrast-enhancing part. CONCLUSIONS: In spite of a small sample size and partly a wide range of values, the authors found statistically significant differences between meningiomas, metastases, and lymphomas concerning ADC values. Nevertheless, a differentiation of these lesions only with the help of ADC values seems questionable. PMID- 16254400 TI - Adjusted scaling of FDG positron emission tomography images for statistical evaluation in patients with suspected Alzheimer's disease. AB - BACKGROUND AND PURPOSE: Statistical parametric mapping (SPM) gained increasing acceptance for the voxel-based statistical evaluation of brain positron emission tomography (PET) with the glucose analog 2-[18F]-fluoro-2-deoxy-d-glucose (FDG) in patients with suspected Alzheimer's disease (AD). To increase the sensitivity for detection of local changes, individual differences of total brain FDG uptake are usually compensated for by proportional scaling. However, in cases of extensive hypometabolic areas, proportional scaling overestimates scaled uptake. This may cause significant underestimation of the extent of hypometabolic areas by the statistical test. METHODS: To detect this problem, the authors tested for hypermetabolism. In patients with no visual evidence of true focal hypermetabolism, significant clusters of hypermetabolism in the presence of extended hypometabolism were interpreted as false-positive findings, indicating relevant overestimation of scaled uptake. In this case, scaled uptake was reduced step by step until there were no more significant clusters of hypermetabolism. RESULTS: In 22 consecutive patients with suspected AD, proportional scaling resulted in relevant overestimation of scaled uptake in 9 patients. Scaled uptake had to be reduced by 11.1% +/- 5.3% in these cases to eliminate the artifacts. Adjusted scaling resulted in extension of existing and appearance of new clusters of hypometabolism. Total volume of the additional voxels with significant hypometabolism depended linearly on the extent of the additional scaling and was 202 +/- 118 mL on average. CONCLUSIONS: Adjusted scaling helps to identify characteristic metabolic patterns in patients with suspected AD. It is expected to increase specificity of FDGPET in this group of patients. PMID- 16254401 TI - Comparison of 1- and 2-marker techniques for calculating system magnification factor for angiographic measurement of intracranial vessels. AB - BACKGROUND AND PURPOSE: Accurate estimation of an intracranial vessel size is crucial during a diagnostic or therapeutic angiography procedure. The use of 1 or 2 external markers of known size is previously proposed to manually estimate the magnification factor (MF) of an intracranial vessel. The authors evaluated the use of different external marker techniques commonly used during angiographic measurements. METHODS: Forty-three intracranial vessels in 17 patients were measured using 1-and 2-marker techniques. To obtain the MF, 2 metallic markers were attached to the frontal-temporal regions. The MFs for the targeted vessels were obtained from the x-ray films by measuring the image sizes of the markers and their positions with respect to the target vessel. RESULTS: Using a phantom, the errors resulted from (a) linear interpolation of MFs, (b) linear interpolation of inverse MFs, and (c) using the MFs of 1 marker, which were 1.23% to 2.23%, 0.8% to 1.55%, and 3.85% to 14.62%, respectively. A similar trend was observed for the measurement of cerebral arteries. CONCLUSION: The use of 2 markers can result in a more accurate estimation of the vessel size. The use of only 1 external marker can lead to substantial error based on the location of the target vessel. Optimizing image acquisition is also crucial for accurate determination of vessel size. PMID- 16254403 TI - Corpus callosum atrophy in Wernicke's encephalopathy. AB - BACKGROUND AND PURPOSE: Neuropathologic changes in Wernicke's encephalopathy (WE) involve variable brain structures. Corpus callosum involvement in WE, however, is largely unknown. The authors investigated the degree and the pattern of corpus callosum changes in WE according to the etiologies. METHODS: Nineteen patients with WE (between 34 and 81 years) and 19 age- and sex-matched control participants were included. The total cross-sectional callosal area and 5 callosal subregions (C1-C5) were measured by tracing outer margins in the midsagittal sections. Subregions were determined by placing radial dividers with 10 rays. The pixel numbers for corpus callosums were calculated, and the values obtained were adjusted for head size variations. RESULTS: The causes of WE were alcoholism (10), intestinal surgery (5), anorexia (3), and hyperemesis gravidarum (1). The mean size of the total corpus callosum was significantly reduced in alcoholic WE (P< .001; 527.8 +/- 70.8 mm2 for alcoholic WE; 664.6 +/- 58.1 mm2 for the corresponding controls), but not in nonalcoholic WE. In subregion analysis, prefrontal callosum (C2) atrophy was the most prominent in alcoholic WE. In contrast, only splenium (C5) was atrophied in nonalcoholic WE. The degree of atrophy did not change throughout the follow-up period (mean 5.3 weeks). CONCLUSION: This study suggests that the extent and location of corpus callosum atrophy differs between alcoholic WE and nonalcoholic WE, implying separate contribution of alcohol neurotoxicity and nutritional deficiency. PMID- 16254402 TI - Magnetic resonance spectroscopy of the thalamus in essential tremor patients. AB - BACKGROUND AND PURPOSE: Although essential tremor (ET) is one of the most common movement disorders, its pathogenesis remains obscure. The ventral intermediate nucleus of the thalamus (VIM nucleus) is suggested to play an important role in the occurrence of disease. In this study, the authors investigated the presence of biochemical or metabolic alterations in the thalamus of patients with ET using magnetic resonance (MR) spectroscopy. METHODS: The study group included 14 patients with ET who suffered from tremor predominantly in their right arm and 9 healthy controls. All patients and controls were right handed. Following conventional cranial MR imaging, single voxel proton MR spectroscopy of the thalamus involving the VIM nuclei was performed bilaterally in both the patients with ET and controls. Metabolite peaks of choline (Cho), creatine (Cr), and Nacetylaspartate (NAA) were obtained from each spectroscopic volume of interest. The right and left thalamic NAA/Cr and Cho/ Cr ratios were compared first within the patient group and then between the control and patient groups. The differences in age and spectroscopic data between groups were assessed using the Mann-Whitney U test, whereas the comparison within groups between left thalamus and right thalamus was done by the Wilcoxon test. RESULTS: In patients with ET, the NAA/Cr ratio of the right thalamus was found to be significantly higher than the NAA/Cr ratio of the left thalamus (P= .02). However, NAA/Cr and Cho/Cr ratios were found to be similar (P> .05) when we compared the control and patient groups for the right thalamus and then the left thalamus. CONCLUSION: These data present preliminary evidence for metabolic alterations of the contralateral thalamus (namely, low NAA/Cr ratio) in ET patients with predominantly involved right arm. However, the series is small and further data are necessary to clear the subject adequately. PMID- 16254404 TI - Restricted diffusion of the splenium in acute Wernicke's encephalopathy. AB - Acute Wernicke's encephalopathy (WE) is caused by profound vitamin B1 (thiamine) deficiency and commonly presents with the classic clinical triad of mental confusion, ataxia, and ophthalmoplegia. This characteristic presentation results from the propensity of acute thiamine deficiency to preferentially injure specific brain regions: the dorsomedial thalamus, periaqueductal gray, and mamillary bodies. In these regions, abnormal magnetic resonance signaling on conventional sequences has been well described; however, diffusion restriction has only recently been reported. The authors demonstrate diffusion-weighted imaging (DWI) abnormalities of the splenium of the corpus callosum in a patient with acute WE, which has not been reported previously, and suggest a potential pathological mechanism. With the recent addition of DWI, MRI is becoming more sensitive to the changes in acute WE. Furthermore, the use of apparent diffusion coefficient mapping to evaluate the extent of likely underlying cytotoxic injury may help determine long-term response to vitamin therapy and, thus, disability. PMID- 16254405 TI - Wyburn-Mason syndrome. PMID- 16254409 TI - Significance of vascular endothelial growth factor expression in renal tissue of patients with preeclamptic nephropathy. AB - OBJECTIVE: It was the aim of this study to evaluate the distribution and expression of vascular endothelial growth factor (VEGF) in kidneys of patients with preeclamptic nephropathy and their relationship with clinical and pathological manifestations. METHODS: From May 1993 to August 2004, 19 patients with a mean age of 28.1 +/- 4.53 years (range 23-40), diagnosed with preeclamptic nephropathy by renal biopsy, were enrolled in this study. Fifteen were nulliparous and 4 multipara. Their renal tissues were subjected to immunohistochemical staining by a four-layer peroxidase-antiperoxidase method using monoclonal anti-VEGF. Residual normal renal tissue obtained at nephrectomy served as control. The relationship between the expression pattern of VEGF and clinicopathological features was also investigated. RESULTS: The expression of VEGF markedly increased in renal tissues of patients with preeclamptic nephropathy at the early stage of gestation termination in comparison with normal controls. However, over time, it gradually decreased and reached the level of normal controls (100 vs. 71.43 vs. 20%, p < 0.05). The degree of endothelial proliferation in the glomeruli was closely related to the expression of VEGF, which was stronger in patients with diffuse endothelial proliferation than in those with segment proliferation (p < 0.05). In addition, there was a proportional relationship between the expression of VEGF and the level of urinary protein excretion (p < 0.05). CONCLUSION: The patients with preeclamptic nephropathy showed strong expressions of VEGF in glomeruli, which were closely associated with glomerular endothelial lesions and proteinuria, and over time, gradually weakened to normal level after gestation termination. PMID- 16254410 TI - Is hepcidin a link between anemia, inflammation and liver function in hemodialyzed patients? AB - BACKGROUND: Hepcidin synthesis in hepatocytes is modulated in response to anemia, hypoxia or inflammation. A cross-sectional study was performed to assess hepcidin correlations with markers of iron status, erythropoietin therapy and markers of inflammation in hemodialyzed patients and in the healthy volunteers. METHODS: Iron status, complete blood count, creatinine, albumin, lipids were assessed using standard laboratory methods. Hepcidin and high-sensitivity CRP were measured using commercially available kits. RESULTS: Serum iron, TIBC, TSAT, erythrocyte count, Hb, Ht, platelet count, albumin, and cholesterol were lower, whereas ferritin and hepcidin were higher in hemodialyzed patients over controls. Hepcidin correlated positively with triglycerides, albumin, aspartate aminotransferase, lymphocyte count, ferritin and erythropoietin dose and negatively with erythrocyte count, Hb, and Ht in hemodialyzed patients. In multiple regression analysis, triglycerides (beta value was 0.28, p = 0.02) and albumin (beta value was -0.31, p = 0.006) were correlates of hepcidin in hemodialyzed patients. CONCLUSIONS: Elevated hepcidin levels in hemodialyzed patients may be due to functional iron deficiency and anemia. Liver plays an important role in the synthesis of hepcidin. Low-grade inflammation, frequently found in hemodialyzed patients, might also contribute to elevated hepcidin concentration. Hypothesis that hepcidin might link anemia, inflammation and liver function in kidney disease should be further evaluated. PMID- 16254411 TI - Hindbrain development and evolution: past, present, and future. PMID- 16254412 TI - Comparisons and homology in adult and developing vertebrate central nervous systems. AB - Comparisons of characters in both adult and developing vertebrate central nervous systems require an understanding of the concept of homology. This article begins with a definition of homology in adult animals and then discusses criteria and methodology used to make appropriate comparisons of characters at a variety of hierarchical levels. Crucial to such an analysis is the methodology employed by neurocladistics to ensure meaningful comparisons. Then, a similar approach is used to address these identical problems in embryos. Concerns unique to comparisons of developing central nervous systems are enumerated. In addition, a number of special features of central nervous system formation and organization in both adults and embryos are discussed within the framework of homology and neurocladistics. Lastly, the concept of field homology as applied to vertebrate central nervous system characters is addressed. PMID- 16254413 TI - Evolutionary patterns of cranial nerve efferent nuclei in vertebrates. AB - All vertebrates have a similar series of rhombomeric hindbrain segments within which cranial nerve efferent nuclei are distributed in a similar rostrocaudal sequence. The registration between these two morphological patterns is reviewed here to highlight the conserved vs. variable aspects of hindbrain organization contributing to diversification of efferent sub-nuclei. Recent studies of segmental origins and migrations of branchiomotor, visceromotor and octavolateral efferent neurons revealed more segmental similarities than differences among vertebrates. Nonetheless, discrete variations exist in the origins of trigeminal, abducens and glossopharyngeal efferent nuclei. Segmental variation of the abducens nucleus remains the sole example of efferent neuronal homeosis during vertebrate hindbrain evolution. Comparison of cranial efferent segmental variations with surrounding intrinsic neurons will distinguish evolutionary changes in segment identity from lesser transformations in expression of unique neuronal types. The diversification of motoneuronal subgroups serving new muscles and functions appears to occur primarily by elaboration within and migration from already established segmental efferent pools rather than by de novo specification in different segmental locations. Identifying subtle variations in segment specific neuronal phenotypes requires studies of cranial efferent organization within highly diverse groups such as teleosts and mammals. PMID- 16254414 TI - Time-lapse analysis reveals a series of events by which cranial neural crest cells reroute around physical barriers. AB - Segmentation is crucial to the development of the vertebrate body plan. Underlying segmentation in the head is further revealed when cranial neural crest cells emerge from even numbered rhombomeres in the hindbrain to form three stereotypical migratory streams that lead to the peripheral branchial arches. To test the role of intrinsic versus extrinsic cues in influencing an individual cell's trajectory, we implanted physical barriers in the chick mesoderm, distal to emerging neural crest cell stream fronts. We analyzed the spatio-temporal dynamics as individual neural crest cells encountered and responded to the barriers, using time-lapse confocal imaging. We find the majority of neural crest cells reach the branchial arch destinations following a repeatable series of events by which the cells overcome the barriers. Even though the lead cells become temporarily blocked by a barrier, cells that follow from behind find a novel pathway around a barrier and become de novo leaders of a new stream. Surprisingly, quantitative analyses of cell trajectories show that cells that encounter an r3 barrier migrate significantly faster but less directly than cells that encounter an r4 barrier, which migrate normally. Interestingly, we also find that cells temporarily blocked by the barrier migrate slightly faster and change direction more often. In addition, we show that cells can be forced to migrate into normally repulsive territory. These results suggest that cranial neural crest cell trajectories are not intrinsically determined, that cells can respond to minor alterations in the environment and re-target a peripheral destination, and that both intrinsic and extrinsic cues are important in patterning. PMID- 16254415 TI - Specification and patterning of neural crest cells during craniofacial development. AB - Craniofacial evolution is considered fundamental to the origin of vertebrates and central to this process was the formation of a migratory, multipotent cell population known as the neural crest. The number of cell types that arise from the neural crest is truly astonishing as is the number of tissues and organs to which the neural crest contributes. In addition to forming melanocytes as well as many neurons and glia in the peripheral nervous system, neural crest cells also contribute much of the cartilage, bone and connective tissue of the face. These multipotent migrating cells are capable of self renewing decisions and based upon these criteria are often considered stem cells or stem cell-like. Rapid advances in our understanding of neural crest cell patterning continue to shape our appreciation of the evolution of neural crest cells and their impact on vertebrate craniofacial morphogenesis. PMID- 16254416 TI - Spinal nerve innervation to the sonic muscle and sonic motor nucleus in red piranha, Pygocentrus nattereri (Characiformes, Ostariophysi). AB - The red piranha, Pygocentrus nattereri, produces sounds by rapid contractions of a pair of extrinsic sonic muscles. The detailed innervation pattern of the sonic muscle of the red piranha was investigated. The sonic muscle is innervated by branches (sonic branches) of the third (S3so), fourth (S4so), and fifth (S5so) spinal nerves. The average total number of nerve fibers contained in the right sonic branches (n = 5; standard length, SL, 71-85 mm) was 151.8 (standard deviation, SD, 28.3). The occipital nerve did not innervate the sonic muscle. The sonic motor nucleus (SMN) in the piranha was identified by tracer methods using wheat germ agglutinin-conjugated horseradish peroxidase; labeled sonic motor neurons were only observed on the side ipsilateral to the sonic muscle injected with the tracer. In the transverse sections, the labeled sonic motor neurons were located in the dorsal zone (mainly large and medium neurons) and in the ventral zone (mainly small neurons) of the ventral horn. In the horizontal sections, the labeled neurons formed a rostrocaudally elongated SMN from the level of the caudal part of the second spinal nerve root to the intermediate region between the fifth and sixth spinal nerve roots. The average number of the labeled neurons (n = 5; SL, 64-87 mm) was 152.6 (SD, 7.3). We conclude that the sonic muscles of the piranha are innervated by approximately 300 sonic motor neurons located only in the spinal cord. PMID- 16254417 TI - Long-term effect of intravenous thrombolytic therapy in acute stroke: responder analysis versus uniform analysis of excellent outcome. AB - BACKGROUND: Knowledge regarding functional improvement over time and long-term outcome after intravenous thrombolysis in acute ischaemic stroke is limited. The aim of this study was to compare a uniform assessment of outcome with an assessment taking the baseline stroke severity into account (responder analysis). METHODS: Fifty-seven patients were assessed with the modified Rankin Scale at 3, 6 and 12 months and a comparison was made between a uniform assessment and a responder analysis of excellent outcome. RESULTS: Between 3 and 12 months, 74% of the patients had a stable functional outcome and 22% improved. Excellent outcome at 12 months was similar in the uniform analysis (37%) and the responder analysis (35%). The individual patients having an excellent outcome differed, however, using the two methods. Using a responder analysis the number of patients with excellent outcome decreased in mild stroke patients by 40%, but increased in severe stroke patients by 43%. CONCLUSIONS: Short-term outcome is sustained at 12 months, but major improvement does not occur between 3 and 12 months. A responder analysis of long-term excellent outcome provided a balanced measure of outcome reflecting the drug-related potential of improvement in all stroke severity subgroups, whereas a uniform analysis provided a measure of outcome mainly in mild stroke patients. These results suggest that a responder analysis should be considered for the assessment of outcome after treatment for acute stroke. PMID- 16254418 TI - Brain embolism caused by a mobile aortic thrombus with iron deficiency anemia. PMID- 16254419 TI - Hemopericardium following intravenous thrombolysis for acute ischemic stroke. PMID- 16254420 TI - Multiple acute ischemic brain lesions and increased fluorodeoxyglucose uptake in the ascending aorta. PMID- 16254421 TI - Mild cyanosis due to coexistence of congenitally corrected transposition of the great arteries and Gerbode-type defect. PMID- 16254422 TI - A simple and effective regimen for prevention of radial artery spasm during coronary catheterization. AB - Radial artery spasm occurs frequently during the transradial approach for coronary catheterization. Premedications with nitroglycerin and verapamil have been documented to be effective in preventing radial spasms. Verapamil is relatively contraindicated for some patients with left ventricular dysfunction, hypotension and bradycardia. We would like to know whether nitroglycerin alone is sufficient for the prevention of radial artery spasm. We conducted a randomized controlled trial to compare the spasmolytic effect between heparin alone, heparin plus nitroglycerin and heparin plus nitroglycerin and varapamil during transradial cardiac catheterization. In this study, a total of 406 patients underwent transradial cardiac catheterization and intervention. After successful cannulation and sheath insertion of radial arteries, 133 patients in group A received 3,000 units of heparin, 100 microg of nitroglycerin and 1.25 mg of verapamil via sheath, 135 patients in group B received 3,000 units of heparin and 100 microg of nitroglycerin, and 93 patients in group C received 3,000 units of heparin. Five patients in group A (3.8%), 6 patients in group B (4.4%) and 19 patients in group C (20.4%) showed radial spasms. There is no statistically significant difference between groups A and B (p = 0.804), but there are strong statistically significant differences between groups A and C (p = 0.001) and groups B and C (p = 0.003). Intra-arterial premedication with 100 microg nitroglycerin and 3,000 units of heparin is effective in preventing radial spasms during transradial cardiac catheterization. PMID- 16254423 TI - Superior vena cava anomalies in the generation of angina pectoris: a report of two cases. AB - Superior vena cava anomalies are rare occurrences caused by variations in the development of the embryonic venous system. Persistent left superior vena cavae are the most common congenital aberrations in the thoracic venous system, with an incidence of 0.3-0.5%, but their association in the absence of a right superior vena cava is extremely rare and scarcely reported. We report two case studies describing persistent left superior vena cavae found with and without a right superior vena cava in patients presenting with chest pain. A discussion regarding superior vena cava abnormalities as well as the etiology, associations, and diagnosis of these unusual entities follows. PMID- 16254424 TI - Balloon valvuloplasty as an initial palliation in the treatment of newborns and young infants with severely symptomatic tetralogy of Fallot. AB - BACKGROUND: Balloon valvuloplasty in infants with symptomatic tetralogy of Fallot (TOF) may increase the pulmonary flow and prompt the growth of pulmonary arteries. METHOD: From 1994 to 2002, percutaneous transluminal balloon valvuloplasty (PTPV) was performed in 22 consecutive newborns and young infants (<3 months of age) with TOF. The indication included severe hypoxemia (systemic oxygen saturation below 75%, 10 cases) and repeated hypoxic spells (12 cases). The age at PTPV was 8-88 days (38 +/- 34, median 27) and the body weight 2-5 kg (3.45 +/- 1.15, median 3). A balloon catheter (4- 7 mm in diameter and 2 cm in length) was used to dilate the pulmonary valve. RESULTS: No major procedure related complications occurred. The systemic oxygen saturation increased significantly (14 +/- 9%). A subsequent palliative shunt operation was avoided in 12 patients (54.5%), but 10 still needed operation (median 11 days after the PTPV). The presence of recurrent hypoxic spells before PTPV was the most important indicator for PTPV failure (p = 0.02). CONCLUSION: PTPV was safe and effective for symptomatic newborns and young infants with TOF, but a palliative shunt operation was still needed due to short effect, especially in those with recurrent spells before the dilation. PMID- 16254425 TI - Electrocardiographic abnormalities in patients with right ventricular dilation due to acute pulmonary embolism. AB - In a study of 190 patients with acute pulmonary embolism, right ventricular dilation was present in 64 (34%), mean age 58 +/- 15 years. The 18 electrocardiographic abnormalities on the 12-lead electrocardiogram had a sensitivity of 8-69%, a specificity of 70-98%, a positive predictive value of 23 69%, a negative predictive value of 64-83%, a likelihood ratio for a positive test of 1.3-4.4, and a likelihood ratio for a negative test of 0.41-1.10 in predicting right ventricular dilation in patients with acute pulmonary embolism. PMID- 16254426 TI - Relationship between delusions and regional cerebral blood flow in Alzheimer's disease. AB - To investigate the association between delusions and cerebral functional deficits in Alzheimer's disease (AD), we evaluated probable AD patients with and without delusions. METHODS: Functional brain imaging was performed by single photon emission computed tomography with technetium-99m-labeled ethyl cysteinate dimer (99mTc-ECD) in 64 AD patients and 76 age-matched normal healthy volunteers. SPECT data were analyzed by statistical parametric mapping. RESULTS: In AD patients, no differences were found in age and cognitive activities between those with (n = 25) and without (n = 39) delusions. Compared with normal healthy volunteers, AD patients had significantly decreased perfusion in the posterior cingulate gyri, precunei, and parietal association cortex. Moreover, in the patients with delusions, perfusion was significantly decreased in the frontal lobe with right side dominance. In the comparison between the patients with and without delusions, the patients with delusions had significantly decreased perfusion in the prefrontal cortex, anterior cingulate gyri, inferior to middle temporal cortices, and parietal cortex of the right hemisphere (p < 0.01). CONCLUSION: The functional deficits in the right hemisphere may be the cause of delusions in AD. PMID- 16254427 TI - Cube drawing performances in normal ageing and Alzheimer's disease: data from the PAQUID elderly population-based cohort. AB - To assess the influence of age and other sociodemographical variables, depressive symptomatology and the occurrence of Alzheimer's disease (AD) on cube drawing, we examined the performance of 858 normal subjects and 17 incident AD patients in the PAQUID population-based study. Cube drawing was first performed on verbal command and, in case of failure, a model of a cube was given to the subjects to copy. Normal subjects had some difficulties in drawing a cube on verbal command, evidenced by 40% failure in the drawing-to-command condition, while only 17% still failed in the drawing-to-copy condition. Multivariate logistic regressions showed that age, gender, and educational level were associated with cube drawing failure in both conditions, but depressive symptomatology was only associated with persistent failure in the drawing-to-copy condition. Seventy-six point five percent of incident AD subjects failed in the drawing-to-command condition and 64.7% did not benefit from the presentation of the model. Analysis showed that persistent failure in the copy condition was the best level of differentiation between normal and AD subjects. PMID- 16254428 TI - Effect of the apolipoprotein E epsilon4 allele on the efficacy and tolerability of galantamine in the treatment of Alzheimer's disease. AB - OBJECTIVE: To investigate the effect of the apolipoprotein E (ApoE) epsilon4 allele on the efficacy and tolerability of galantamine treatment. METHODS: A total of 202 patients with mild to moderate Alzheimer's disease participated in a 16-week, prospective, multi-center, randomized, double-blind galantamine trial in a Korean population. Patients were assessed at baseline and after 4, 8 and 16 weeks of randomized treatment using the 11-item cognitive subscale of the Alzheimer's Disease Assessment Scale (ADAS-cog/11), the Clinician's Interview Based Impression of Change plus Caregiver Input (CIBIC-plus), the Disability Assessment for Dementia Scale (DAD), the Behavioural Pathology in Alzheimer's Disease Rating Scale (BEHAVE-AD) and adverse events. ApoE genotypes were determined for all subjects. RESULTS: Of the 202 subjects, 115 carried at least one ApoE epsilon4 allele and 87 did not. In both ApoE epsilon4 carriers and ApoE epsilon4 noncarriers, significant improvements were detected relative to baseline on ADAS-cog/11, CIBIC-plus, DAD and BEHAVE-AD. ApoE epsilon4 noncarriers showed better improvement in mean total BEHAVE-AD score and mean psychosis (delusions and hallucinations) subscale score than ApoE epsilon4 carriers. The incidence of weight loss was significantly higher in ApoE epsilon4 carriers (n = 11; 9.6%) than in ApoE epsilon4 noncarriers (n = 1; 1.2%) during this 16-week study, even though 92% of patients who complained of weight loss completed this 16-week trial successfully. CONCLUSION: ApoE epsilon4 genotype does not affect galantamine related improvements in cognition, global rating, function and behavior. Longer prospective studies with larger patient populations are required to confirm these new findings. PMID- 16254429 TI - The clinical dementia rating sum of box score in mild dementia. AB - BACKGROUND: Making an early diagnosis of dementia is becoming increasingly important, but is difficult in practice. The Clinical Dementia Rating (CDR) scale is a widely used dementia staging instrument, yielding a global score and a summated score (sum of box score). This study examines the utility of the CDR sum of box score, rather than the CDR global score, in making a diagnosis of early dementia. OBJECTIVE: To determine whether the CDR sum of box score is predictive of an ICD-10 diagnosis of dementia in cases with mild cognitive deficits. METHODS: Clinical data recorded on our Memory Clinic database were examined for all patients seen over a 6-year period. Data were extracted from 276 first visits in which patients had scored 0.5 using the CDR global score. We examined the relationship between CDR sum of box score and consensus diagnosis of dementia using logistic regression. RESULTS: We found that increased CDR sum of box score was significantly associated with a higher probability of being assigned an ICD 10 diagnosis of dementia (p < 0.001). The odds ratio for the coefficient of CDR sum of box was 2.3 (95% CI 1.7-3.1), indicating that the likelihood of being diagnosed as having dementia increased by a factor of 2.3 for every point increase on the CDR sum of box score. CONCLUSION: These findings indicate that the CDR sum of box score provides additional information to the CDR global score in mild cases. The CDR sum of box score is a helpful indicator in making/excluding a diagnosis of dementia in people with mild cognitive deficits. PMID- 16254430 TI - Clinical and neuroimaging correlates of mild cognitive impairment in a middle aged community sample: the personality and total health through life 60+ study. AB - This cross-sectional study aimed at determining the clinical and structural brain magnetic resonance imaging correlates of mild cognitive impairment (MCI). The data presented here are from the first wave of the longitudinal Personality and Total Health through Life 60+ project. 2,551 community-dwelling individuals in the age range of 60-64 years were recruited randomly through the electoral roll. They were screened using Mini-Mental State Examination and a short cognitive battery. Those who screened positive underwent detailed medical and neuropsychological assessments. Of the 224 subjects who screened positive, 117 underwent a detailed assessment. Twenty-nine subjects fulfilled the Mayo Clinic criteria for MCI. Magnetic resonance imaging scans were analyzed for 26 subjects with MCI as well as normal controls. Subjects were clinically evaluated for depressive symptoms and major and minor depression syndromes. Logistic regression analysis was performed predicting MCI from anterior and mid-ventricular brain ratios, cortical atrophy measures, hippocampal volumes, volumes of amygdala and white matter hyperintensities after adjusting for age, gender, years of education, depression and physical disability. None of the neuroanatomical substrates appeared as predictors of MCI. The only predictors were higher depression scores and fewer years of education. Structural neuroimaging may not have an added advantage in the detection of MCI in middle-aged community-dwelling subjects. It may be that this age group is too young for such brain changes to be identified. PMID- 16254431 TI - Adjuvant 5-FU-based chemoradiotherapy for patients undergoing R-1/R-2 resections for pancreatic cancer. AB - BACKGROUND: Pancreatic cancer is the fifth leading cause of cancer-related death worldwide. Among patients treated with surgery alone, liver metastasis occurs in up to 50%, peritoneal recurrence in 25%, and local recurrence occurs in 50-80% of all patients who underwent resection. Even after a macroscopically curative resection, tumour cells might be observed by microscopy at one or more edges of the resected specimen in 20-51% (R-1) which might account for the high local recurrence. AIM OF THE STUDY: An analysis was performed in 54 patients who underwent an irradical resection (R-1 and R-2) for pancreatic cancer. 33 patients were treated with chemoradiotherapy. To evaluate the effect of therapy on survival and recurrence, this group was retrospectively compared to a group of 21 patients that did not receive chemoradiotherapy. METHODS: Radiotherapy consisted of 50 Gy external upper abdomen radiation in two courses of 3 weeks, concomitant with intravenous 5-FU 25 mg/kg/24 h continuously on the first 4 days of each treatment course. Follow-up was performed mainly by CT scanning and occasionally by US and was completed for all but 1 patient. RESULTS: The treatment protocol was completed in all patients without complications. Local recurrence was found in 6 (18%) patients in the group of patients who received adjuvant therapy versus 16 (48%) patients in the group that did not receive adjuvant therapy (p = 0.001). The median survival time for the treated group was 12.8 vs. 13.7 months in the group that did not receive chemoradiotherapy (p = 0.9). Three (9%) patients are still alive 140, 88 and 70 months after receiving surgery and adjuvant treatment. CONCLUSION: Adjuvant chemoradiotherapy clearly gives a significant better local control. However, treatment with 5-FU and radiotherapy does not improve survival due to distant metastases. This therapy probably prolongs survival in only a few patients. More effective treatment methods have to be designed to prevent metastatic disease and improve survival. PMID- 16254432 TI - Immunogenetics of type 1 diabetes. AB - The T-cell mediated autoimmune process that destroys pancreatic beta cells in type 1 diabetes (T1D) is a complex phenotype influenced by multiple genetic and environmental factors. Human leukocyte antigen (HLA) accounts for about half of the genetic susceptibility, through a large variety of protective and predisposing haplotypes. Other important loci associated with T1D, with much smaller effects than HLA, include the insulin variable number of tandem repeats, PTPN22, and CTLA-4. Detecting the association and confirming it beyond doubt is only the first step. Identifying the functional variant from among a block of polymorphisms in tight linkage disequilibrium and determining its biological consequences can be an even more challenging task. It is hoped that the identification of additional loci and functional analysis of known ones, no matter how small each individual effect is, will provide: (1) pathophysiological insights necessary for the development of preventive interventions; (2) risk prediction to identify individuals that can benefit from them, and (3) potentially, identification of distinct subgenotypes, with different immune dysregulation pathways leading to the common disease phenotype that may respond to different preventive interventions. PMID- 16254433 TI - Clinical features, diagnosis and molecular studies of familial central diabetes insipidus. AB - BACKGROUND: Familial central diabetes insipidus (DI) is rare and is characterised by polydipsia and polyuria with a variable age of onset. The evaluation of arginine vasopressin (AVP) secretion in these individuals has been reported infrequently and only in adulthood. OBJECTIVE: To describe the clinical features, diagnosis and molecular investigation of children affected by familial central DI. METHODS: Functional studies of AVP secretion were undertaken in children from two kindreds with familial central DI. The AVP-neurophysin II (AVP-NPII) gene was also sequenced in symptomatic individuals. RESULTS: In affected individuals, the result of the water deprivation test may be inconclusive. However, the hypertonic saline test identified both the severe and partial forms of AVP deficiency. A novel mutation of the AVP-NPII gene was identified by direct gene sequencing in both families. CONCLUSIONS: This report highlights the progressive decline in AVP secretion with increasing age in this disorder and the usefulness of mutational analysis in these families. In symptomatic individuals, the hypertonic saline test may be a useful second-line investigation for functional studies of AVP secretion where molecular diagnostics are unavailable. PMID- 16254434 TI - Free androgen index is superior to total testosterone for short-term assessment of the gonadal axis after renal transplantation. AB - OBJECTIVE: Recent studies have assessed gonadal function in association with different immunosuppressive drugs in transplanted patients mainly relying on the measurement of total testosterone. It is the aim of this study to assess the short-term changes of the hypothalamic-pituitary-gonadal axis following renal transplantation using the free androgen index (FAI). PATIENTS AND METHODS: The sequential changes in total testosterone, sex hormone-binding globulin (SHBG), gonadotropin and prolactin concentrations were measured in 22 male patients before and after 1-3 days, and 1, 2 and 3 weeks following renal transplantation. RESULTS: Total testosterone and SHBG concentrations dropped significantly after transplantation (total testosterone: baseline: 15.2 +/- 1.6 nmol/l vs. 1 week: 7.9 +/- 0.8 nmol/l vs. 2 weeks: 9.8 +/- 0.9 nmol/l, SHBG: baseline: 29.9 +/- 3.2 nmol/l vs. 1 week: 19.9 +/- 2.1 nmol/l, 2 weeks: 18.9 +/- 2.4 nmol/l, p < 0.01). FAI decreased significantly after day 1-3 returning to values near baseline thereafter (baseline: 60 +/- 9% vs. day 1-3: 38 +/- 6%, 2 weeks: 61 +/- 7%; p < 0.01). There was a significant positive correlation between FAI and renal function. CONCLUSION: Measurement of the free androgen index is superior to total testosterone for assessment of the pituitary-gonadal axis in the first weeks after renal transplantation. PMID- 16254435 TI - Susceptibility alleles and haplotypes of human leukocyte antigen DRB1, DQA1, and DQB1 in autoimmune polyglandular syndrome type III in Japanese population. AB - BACKGROUND: It has been reported that HLA class II haplotypes DRB1*0405-DQA1*0303 DQB1*0401 and DRB1*0901-DQA1*0302-DQB1*0303 are major susceptibility haplotypes for type 1 diabetes mellitus (DM) in Japanese population. However, little has been reported on the susceptibility HLA class II haplotypes in Japanese patients with autoimmune polyglandular syndrome type II and type III (APS III). PATIENTS AND METHODS: HLA class II haplotypes of DRB1-DQA1-DQB1 in 31 patients with APS III, 14 patients with Hashimoto's thyroiditis alone, and 15 patients with Graves' disease alone were examined in Japanese population. APS III patients were divided into three groups (A, B, and C) depending on the combination of autoimmune endocrine diseases. RESULTS: In 13 APS III patients with both Hashimoto's thyroiditis and type 1 DM (group A), the haplotype frequencies of the HLA DRB1*0802-DQA1*0401-DQB1*0402 and DRB1*0901-DQA1*0302-DQB1*0303 were significantly higher than in the controls. In patients with Hashimoto's thyroiditis alone, the haplotype frequency of DRB1*0901-DQA1*0302-DQB1*0303 was significantly higher than in controls, whereas the frequency of DRB1*0802 DQA1*0401-DQB1*0402 did not differ significantly from those in the controls. In 11 APS III patients with both Graves' disease and type 1 DM (group B), the haplotype frequencies of HLA DRB1*0405-DQA1*0303-DQB1*0401 and DRB1*0802 DQA1*0301-DQB1*0302 were significantly higher than in controls. In patients with Graves' disease alone, the haplotype frequency of DRB1*0803-DQA1*0103-DQB1*0601 were significantly higher than those in controls, suggesting that the susceptibility haplotypes for group B APS III differed from those for Graves' disease alone. In 7 APS III patients with both autoimmune thyroid diseases and pituitary disorders (group C), the haplotype frequency of HLA DRB1*0405-DQA1*0303 DQB1*0401 was significantly higher than in controls. CONCLUSIONS: Susceptible HLA class II haplotypes of DRB1-DQA1-DQB1 for APS III differ between the Japanese and Caucasian populations. More interestingly, the susceptible HLA class II haplotypes differ among the three types of Japanese APS III and are not merely a combination of susceptibility haplotypes of each endocrine disease. PMID- 16254436 TI - Short-term secretory regulation of ghrelin during growth hormone provocative tests in prepubertal children with various growth hormone secretory capacities. AB - BACKGROUND AND OBJECTIVE: Ghrelin is a novel gastric peptide which stimulates GH secretion and has been demonstrated to have orexigenic and adipogenic properties. Insulin is a physiological and dynamic modulator of plasma ghrelin, and insulinemia possibly mediates the effect of the nutritional state on the plasma concentrations of ghrelin in adults. No data on the regulation of GH secretion by ghrelin have so far been reported, nor has the possible influence of hypoglycemia on the plasma ghrelin levels in children been reported. METHODS: Provocative studies were performed using a variety of stimuli, including insulin-induced hypoglycemia, and glucagon, arginine and L-dopa loading. We studied a group of 27 children with short stature being investigated for GH deficiency (10 F, 17 M; age 4-14 years; height SDS -0.92 to -3.27); the subjects were instructed to fast overnight, and the following morning, the relationships among the plasma ghrelin, GH and glucose levels were investigated by determining the plasma ghrelin profiles during those provocative tests. Using a new method for determining the two types of ghrelin, samples were obtained for determination of the plasma ghrelin, serum glucose and serum GH levels after the administration of the aforementioned stimulating agents. RESULTS: All the four stimuli caused a significant decrease in the circulating C- and N-ghrelin levels with a nadir at +30 min, with the exception of the N-ghrelin level following the L-dopa loading. During the same period, the plasma GH level increased following insulin, arginine and L-dopa loading, and the plasma glucose level increased significantly following glucagon loading. In the arginine and L-dopa load connected, a significant correlation was observed between the 30-min change in the serum GH level and the 30-min change in the plasma C-ghrelin level. In the multiple regression analysis to explain the 30-min change in the plasma level of C ghrelin, the baseline plasma level of C-ghrelin (basal), height and % overweight were the only three significant parameters, accounting for 85.2% of the variance. CONCLUSION: This study demonstrated that the inverse relation between the circulating GH and ghrelin levels may indicate the existence of a feedback loop, and also lends support to the assumption of a GH-independent relationship between plasma ghrelin and glucose levels. These observations constitute further evidence to suggest that peripheral ghrelin is a direct growth-promoting hormone. PMID- 16254437 TI - The spectrum of allergens in ragweed and mugwort pollen. AB - Ragweed and mugwort are important allergenic weeds belonging to the Asteraceae or Compositae plant family. Pollen of mugwort is one of the main causes of allergic reactions in late summer and autumn in Europe and affects about 10-14% of the patients suffering from pollinosis. Ragweed pollen represents the major source of allergenic protein in the United States, with a prevalence of about 50% in atopic individuals. In Europe, ragweed allergy is now rapidly increasing particularly in certain areas in France, Italy, Austria, Hungary, Croatia, and Bulgaria. Amb a 1 and Art v 1, the major allergens of ragweed and mugwort, respectively, are unrelated proteins. Amb a 1 is an acidic 38-kDa nonglycosylated protein. The natural protein undergoes proteolysis during purification and is cleaved into a 26-kDa alpha chain, which associates noncovalently with the beta chain of 12 kDa. The two-chain form seems to be immunologically indistinguishable from the full length molecule. Art v 1 is a basic glycoprotein comprising two domains: an N terminal cysteine-rich, defensin-like domain and a C-terminal proline/hydroxyproline-rich module. The proline/hydroxyproline-rich domain was recently shown to contain two types of glycosylation: (1) a large hydroxyproline linked arabinogalactan composed of a short beta1,6-galactan core substituted by a variable number (5-28) of alpha-arabinofuranose residues forming branched side chains with 5-, 2,5-, 3,5-, and 2,3,5-substituted arabinoses, and (2) single and adjacent beta-arabinofuranoses linked to hydroxyproline. As described for other pollen, ragweed and mugwort pollen also contain the pan-allergen profilin and calcium-binding proteins, which are responsible for extensive cross-reactivity among pollen-sensitized patients. PMID- 16254438 TI - Activated human eosinophils. PMID- 16254439 TI - Vasoconstrictive effect of angiotensin IV in isolated rat basilar artery independent of AT1 and AT2 receptors. AB - The effect of angiotensin IV (AngIV) was studied in freshly isolated rat basilar arteries (BAs) perfused at a constant rate. AngIV had no effect on basal BA perfusion pressure, but induced a marked concentration-dependent contraction in vessels precontracted by a 50-mM KCl solution (EC50=44.5+/-16 nM). This contraction was unaffected by the angiotensin AT1 receptor antagonist candesartan or the angiotensin AT2 receptor blocker PD123319, but was markedly inhibited by two different specific AT4 receptor antagonists, Nle1-Leu3 yen(CH2-NH2)3-4-AngIV and divalinal-AngIV. Removal of the endothelium abolished the contractile response to AngIV, and pretreatment of endothelium-intact arteries with the endothelin ETA/ETB receptors inhibitor PD142893 blocked the AngIV-induced contraction to the same extent. In BA pretreated with endothelin-1 (ET-1; 0.01 microM), AngIV-induced a concentration-dependent contraction, shifted to the left, compared with that observed with KCl precontraction, unaffected by candesartan but completely abolished by Nle1-Leu3 yen(CH2-NH2)3-4-AngIV. The contractile effect was not affected by endothelium removal in the presence of exogenous ET-1, in contrast to KCl pretreated BA, suggesting that endothelium was mandatory to unmask the effect of AngIV as a source of endogenous ET-1 release. Taken together, these results indicate that low (nanomolar) concentrations of AngIV exert a constrictive effect mediated by its specific binding site AT4 in the rat BA, and that this vasoactive effect is indirect and involves endogenous endothelin(s). PMID- 16254441 TI - Complete inventory of ABC proteins in human pathogenic yeast, Candida albicans. AB - The recent completion of the sequencing project of the opportunistic human pathogenic yeast, Candida albicans (http://www.ncbi.nlm.nih.gov/), led us to analyze and classify its ATP-binding cassette (ABC) proteins, which constitute one of the largest superfamilies of proteins. Some of its members are multidrug transporters responsible for the commonly encountered problem of antifungal resistance. TBLASTN searches together with domain analysis identified 81 nucleotide-binding domains, which belong to 51 different putative open reading frames. Considering that each allelic pair represents a single ABC protein of the Candida genome, the total number of putative members of this superfamily is 28. Domain organization, sequence-based analysis and self-organizing map-based clustering led to the classification of Candida ABC proteins into 6 distinct subfamilies. Each subfamily from C. albicans has an equivalent in Saccharomyces cerevisiae suggesting a close evolutionary relationship between the two yeasts. Our searches also led to the identification of a new motif to each subfamily in Candida that could be used to identify sequences from the corresponding subfamily in other organisms. It is hoped that the inventory of Candida ABC transporters thus created will provide new insights into the role of ABC proteins in antifungal resistance as well as help in the functional characterization of the superfamily of these proteins. PMID- 16254442 TI - Molecular cloning and biochemical characterization of L-N-carbamoylase from Sinorhizobium meliloti CECT4114. AB - An N-carbamoyl-L-amino acid amidohydrolase (L-N-carbamoylase) from Sinorhizobium meliloti CECT 4114 was cloned and expressed in Escherichia coli. The recombinant enzyme catalyzed the hydrolysis of N-carbamoyl alpha-amino acid to the corresponding free amino acid, and its purification has shown it to be strictly L specific. The enzyme showed broad substrate specificity, and it is the first L-N carbamoylase that hydrolyses N-carbamoyl-L-tryptophan as well as N-carbamoyl L amino acids with aliphatic substituents. The apparent Km values for N-carbamoyl-L methionine and tryptophan were very similar (0.65 +/- 0.09 and 0.69 +/- 0.08 mM, respectively), although the rate constant was clearly higher for the L-methionine precursor (14.46 +/- 0.30 s(-1)) than the L-tryptophan one (0.15 +/- 0.01 s(-1)). The enzyme also hydrolyzed N-formyl-L-methionine (kcat/Km = 7.10 +/- 2.52 s(-1) x mM(-1)) and N-acetyl-L-methionine (kcat/Km = 12.16 +/- 1.93 s(-1) x mM(-1)), but the rate of hydrolysis was lower than for N-carbamoyl-L-methionine (kcat/Km = 21.09 +/- 2.85). This is the first L-N-carbamoylase involved in the 'hydantoinase process' that has hydrolyzed N-carbamoyl-L-cysteine, though less efficiently than N-carbamoyl-L-methionine. The enzyme did not hydrolyze ureidosuccinic acid or 3 ureidopropionic acid. The native form of the enzyme was a homodimer with a molecular mass of 90 kDa. The optimum conditions for the enzyme were 60 degrees C and pH 8.0. Enzyme activity required the presence of divalent metal ions such as Ni2+, Mn2+, Co2+ and Fe2+, and five amino acids putatively involved in the metal binding were found in the amino acid sequence. PMID- 16254443 TI - Increasing PCR fragment stability and protein yields in a cell-free system with genetically modified Escherichia coli extracts. AB - Escherichia coli cell-free protein synthesis is a highly productive system that can be applied to high throughput expression from polymerase chain reaction (PCR) products in 96-well plates for proteomic studies as well as protein evolution. However, linear DNA instability appears to be a major limitation of the system. We modified the genome of the E. coli strain A19 by removing the endA gene encoding the endonuclease I and replacing the recCBD operon (in which recD encodes the exonuclease V) by the lambda phage recombination system. Using the cell extract from this new strain increased the stability of PCR products amplified from a plasmid containing the cat gene. This resulted in CAT (chloramphenicol acetyltransferase) production from PCR products comparable to that from plasmids (500-600 microg/ml) in a batch reaction. We show that cell free protein synthesis reactions using PCR products amplified from genomic DNA and extended with the T7 promoter and the T7 terminator give the same high yields of proteins (550 microg/ml) in 96-well plates. With this system, it was possible to rapidly express a range of cytoplasmic and periplasmic proteins. PMID- 16254444 TI - Quantitative effects of carbohydrates and aromatic amino acids on Clostridium botulinum toxin gene expression using a rapid competitive RT/PCR assay. AB - A rapid competitive RT/PCR assay was developed to determine the effects of nutrients on Clostridium botulinum type E toxin gene expression. The type E strain (EVH) was grown in a nutrient-rich broth containing 1% glucose (base medium). Toxin gene expression was quantified at both mid and late exponential phases of growth. It was found that toxin encoding mRNA levels were highly growth phase dependent with elevated levels found in late exponential phase compared to mid exponential phase. Changing the carbohydrate source had a smaller effect on toxin encoding mRNA levels but as earlier results have suggested, toxin encoding mRNA levels show a strong correlation with type E growth rate. The results have important implications for the food industry whereby risk of type E botulism could be correlated to the nutrient composition of the contaminated food or assessed from C. botulinum growth rates in challenged foodstuffs. PMID- 16254445 TI - Isolation of bacteriophages from Bartonella vinsonii subsp. berkhoffii and the characterization of Pap31 gene sequences from bacterial and phage DNA. AB - Bacteriophages enhance bacterial survival, facilitate bacterial adaptation to new environmental conditions, assist in the adaptation to a new host species, and enhance bacterial evasion or inactivation of host defense mechanisms. We describe the detection and purification of a novel tailed bacteriophage from Bartonella vinsonii subsp. berkhoffii, which was previously described as a bacteriophage negative species. We also compare B. vinsonii subsp. berkhoffi Pap31 bacteriophage gene sequences to B. henselae (Houston I), and B. quintana (Fuller) bacteriophage Pap31 sequences. Negative staining electron microscopy of log phase culturesof B. vinsonii subsp. berkhoffii identified bacteriophages, possessing a 50-nm icosahedric head diameter and a 60- to 80-nm contractile tail. Sequence analysis of the bacteriophage Pap31 gene from B. vinsonii subsp. berkhoffii showed three consensus sequences and a 12-bp insertion when compared with Pap31 gene sequences from B. henselae (Houston I) and B. quintana (Fuller) bacteriophages. Isolation of B. vinsonii subsp. berkhoffii bacteriophages containing a Pap31 gene suggests that this heme-binding protein gene might play an important role in bacterial virulence through the genetic exchange of DNA within this subspecies. Defining phage-associated genes may also contribute to the enhanced understanding of the evolutionary relationships among members of the genus Bartonella. PMID- 16254447 TI - The Streptomyces lividans cytoplasmic signal recognition particle receptor FtsY is involved in protein secretion. AB - The bacterial version of the mammalian signal recognition particle (SRP) and its receptor alpha-subunit (FtsY) is well conserved and essential to all known bacteria. In gram-negative bacteria, the SRP pathway mediates a co-translational targeting of most inner membrane proteins. Additionally, in Streptomyces lividans, a gram-positive bacterium, SRP also targets secretory proteins to the translocon. The role of S. lividans FtsY has been assessed in this work. Co immunoprecipitation studies confirmed that FtsY is associated with the S. lividans SRP in the cytoplasm and that this complex also co-immunoprecipitated with pre-agarase, suggesting that the SRP receptor is involved in SRP-mediated targeting of secretory proteins in S. lividans. Furthermore, the SRP remains attached for the most part to the cellular membrane when the cleavage of pre secretory proteins is severely reduced in a strain lacking the gene coding for the major type-I signal peptidase. PMID- 16254446 TI - Heterologous aquaporin (AQY2-1) expression strongly enhances freeze tolerance of Schizosaccharomyces pombe. AB - Aquaporin membrane proteins enable the transport of water across membranes in various organisms. In yeast their expression has been shown to correlate strongly with freeze tolerance. When we analyzed the freeze tolerance of Schizosaccharomyces pombe, an organism whose genome sequence has revealed no genes encoding a bona fide water channel, we found very low intrinsic freeze tolerance compared to other yeast species with aquaporin-encoding genes. Deletion of Spac977.17, which encodes a putative glycerol facilitator, resulted in no significant differences in freeze tolerance with its corresponding wild-type strain in all growth conditions tested. However, when we expressed the Saccharomyces cerevisiae aquaporin-encoding gene AQY2-1 in S. pombe cells, we found that the relatively low freeze tolerance of S. pombe could be significantly enhanced. Therefore, (i) the absence of a bona fide water channel in S. pombe might provide in part an explanation for its overall low freeze tolerance compared to other yeast species, and (ii) aquaporin overexpression might be a tool to improve cryopreservation of many other cell types as well, as has recently been shown for mouse oocytes and fish embryos. PMID- 16254448 TI - Multiple sclerosis in Latin America. PMID- 16254449 TI - Change in cognitive function in Alzheimer's disease in African-American and white persons. AB - Few studies have examined the association of race with change in cognitive function in Alzheimer's disease (AD). We studied the rate of decline in global and specific measures of cognitive function in a cohort of 410 older African Americans and whites with clinically diagnosed AD. Persons were examined annually for an average of 3.5 years, and follow-up participation among survivors exceeded 90%. In mixed-effects models that controlled for age, gender, education, and premorbid reading activity, African-Americans scored lower than whites at baseline on a composite measure of global cognition and on specific measures of visuoconstruction and naming. However, they experienced less rapid decline in episodic memory (p < 0.01), with similar but not quite significant effects for global cognition (p = 0.06), perceptual speed (p = 0.07) and naming (p = 0.08). The results suggest that the rate of cognitive decline in AD is slower in African Americans compared with whites, particularly for episodic memory. PMID- 16254450 TI - Regional variability in the prevalence of cerebral white matter lesions: an MRI study in 9 European countries (CASCADE). AB - White matter lesions (WML) on MRI of the brain are common in both demented and nondemented older persons. They may be due to ischemic events and are associated with cognitive and physical impairments. It is not known whether the prevalence of these WML in the general population differs across European countries in a pattern similar to that seen for coronary heart disease. Here we report the prevalence of WML in 1,805 men and women drawn from population-based samples of 65- to 75-year-olds in ten European cohorts. Data were collected using standardized methods as a part of the multicenter study CASCADE (Cardiovascular Determinants of Dementia). Centers were grouped by region: south (Italy, Spain, France), north (Netherlands, UK, Sweden), and central (Austria, Germany, Poland). In this 10-year age stratum, 92% of the sample had some lesions, and the prevalence increased with age. The prevalence of WML was highest in the southern region, even after adjusting for differences in demographic and selected cardiovascular risk factors. Brain aging leading to disabilities will increase in the future. As a means of hypothesis generation and for health planning, further research on the geographic distribution of WML may lead to the identification of new risk factors for these lesions. PMID- 16254451 TI - Relation of blood pressure to risk of incident Alzheimer's disease and change in global cognitive function in older persons. AB - PURPOSE: To examine the relation of systolic and diastolic blood pressure to incident Alzheimer's disease (AD) and rate of cognitive change. METHODS: Longitudinal cohort study with annual clinical evaluations. At baseline, blood pressure was measured, apolipoprotein E (APOE) genotyping was performed, and medications were reviewed. RESULTS: 824 older Catholic clergy members without baseline dementia were recruited from across the United States. During a mean of about 6 years of observation, 151 persons developed AD. In a proportional hazards model adjusted for age, sex and education, neither systolic (relative risk = 0.995; 95% CI: 0.986, 1.004, p = 0.249) nor diastolic (relative risk = 1.000; 95% CI: 0.985, 1.015, p = 0.975) blood pressure was related to AD incidence. In mixed effects models, neither systolic nor diastolic blood pressure was related to level or to annual rate of change on a global measure of cognition. These results did not change in subsequent models that accounted for the use of medications with antihypertensive properties or for the possession of an APOE epsilon4 allele. CONCLUSIONS: In a cohort of older persons with a majority taking medications with antihypertensive properties, we did not find a relationship between blood pressure and risk of AD or cognitive decline. PMID- 16254452 TI - Chronic meningitis in Thailand. Clinical characteristics, laboratory data and outcome in patients with specific reference to tuberculosis and cryptococcosis. AB - The charts of 114 consecutive patients with chronic meningitis admitted to a general hospital in Bangkok, Thailand, between 1993 and 1999 were retrospectively reviewed. The most common causative agents were Cryptococcus neoformans (54%) and Mycobacterium tuberculosis (37%). HIV and other underlying diseases had a major impact on the presentation of chronic cryptococcal meningitis patients. Compared to HIV-negative cryptococcal meningitis patients (21%), HIV-positives (79%) had a significantly lower incidence of focal signs (p = 0.02), hydrocephalus (p = 0.03) and seizures (p = 0.001) during hospital stay, furthermore, a lower leucocyte level, a significantly higher glucose level (p = 0.02) and a lower protein level (p = 0.03) in the first cerebrospinal fluid examination. Of the 43 patients with chronic tuberculous meningitis, only 3 were HIV positive. Focal neurologic deficits were found more frequently in tuberculous meningitis patients (p = 0.001) when compared to cryptococcal meningitis patients without HIV. Cerebral infarction on cerebral CT was indicative of tuberculous meningitis. Cryptococcal meningitis patients with HIV infection had a worse outcome compared to non-AIDS patients. Advanced stage of the disease on admission, decreased level of consciousness prior to and on the admission day and raised intracranial pressure above 40 cm H(2)O at any given time were predictive of a poor outcome in tuberculous meningitis patients. PMID- 16254453 TI - Plasma total homocysteine levels in stroke patients screened for the vitamin intervention for stroke prevention clinical trial in the era of folate fortification. AB - Folic acid fortification of grain products was mandated in the USA by January 1998 and in Canada by November 1998. It was hypothesized that screening total plasma homocysteine levels adjusted for age, sex, race and country that were drawn in stroke patients for the Vitamin Intervention for Stroke Prevention trial from 1997 to 2001 would be steady when fortification was completed. Samples were grouped by years 1997/1998, 1999 and 2000/2001, and adjusted means were calculated using a general linear regression model. In 2,612 US and 1,059 Canadian patients, levels showed no consistent trend in the USA alone, while Canadian levels declined (p = 0.06 overall, 0.0003 in the oldest age group). US levels were 0.39 micromol/l (95% CI: -0.08, 0.85) lower than in Canada. Neither fasting nor time since stroke affected the results. PMID- 16254454 TI - Quantitative measures of gait characteristics indicate prevalence of underlying subclinical structural brain abnormalities in high-functioning older adults. AB - Abnormal gait in high-functioning older adults may indicate underlying subtle structural brain abnormalities. We tested the hypothesis that temporal and spatial parameters of gait, including speed, stride length and double support time, are cross-sectionally associated with white matter hyperintensity, subcortical infarcts or brain atrophy on brain MRI. We examined 321 men and women (mean age = 78.3) participating to the Cardiovascular Health Study who were free of dementia or stroke at the time of the gait assessment. Analyses were set with gait as independent variable and brain MRIs as dependent variables. Gait measures were determined from the footfalls recorded on a 4-meter-long instrumented walking surface, the GaitMat II. Brain MRIs were examined for the presence of white matter hyperintensity (WMG, graded from 0 to 9), brain infarcts (predominantly subcortical) and ventricular enlargement (graded from 0 to 9). Slower gait, shorter stride length and longer double support times were associated with greater prevalence of white matter grade > or =3 (p = 0.02), and at least 1 brain infarct (p = 0.04) independent of age. In multivariate logistic regression models adjusted for demographics and clinical cardiovascular diseases, those with gait speed <1.02 m/s were more likely to have WMG > or =3 and at least 1 brain infarct, compared with those with faster gait - odds ratio (OR): 2.85, 95% confidence interval (95% CI): 1.35, 6.02, and OR: 2.09, 95% CI: 1.04, 4.19. Shorter stride length was also associated with greater probability of having at least 1 brain infarct (gait stride <0.88 vs. >1.10 m: OR: 3.20, 95% CI: 1.49, 6.88), while longer double support times were associated with a greater probability of having WMG > or =3 (double support time >0.19 vs. <0.14 s: OR: 2.3, 95% CI: 1.1, 4.7) independent of demographics and clinical cardiovascular diseases. Gait parameters were not significantly associated with ventricular grade. In summary, in this group of high-functioning older adults, poorer gait speed, shorter stride and longer double support time are associated with high white matter disease and subclinical strokes. PMID- 16254455 TI - Epidemiology of oral cavity cancer in taiwan with emphasis on the role of betel nut chewing. AB - This article reports the epidemiological characteristics and the possible contributing etiology of oral cavity cancer in Taiwan. Data on oral cavity cancer from the period between 1986 and 1997 were compiled from the Taiwan Cancer Registry Annual Report. The amount of average annual consumption per person of cigarettes, alcohol and betel nut were extracted from the Annual Report of Taiwan Tobacco and Wine Monopoly Bureau and the Agriculture Counsel of Taiwan. The incidence of oral cavity cancer increased annually. Both the total and male incidence have increased substantially since 1993. Regarding the peak incidence, most cases were seen in the sixth to eighth decades of life. Multiple regression models indicated that 86.2% variation in the incidence of oral cavity cancer was explained by the annual average betel nut consumption per person. These results imply that those who chew betel nut belong to a high-risk group and require special consideration and attention regarding health education and health promotion. PMID- 16254456 TI - Differential effects of fexofenadine on arachidonic acid metabolism in cultured human monocytes. AB - The relief of nasal congestion with the antihistamine fexofenadine in seasonal allergic rhinitis is thought to be due to its additional anti-inflammatory properties. The objective of this study was to evaluate the in vitro effects of fexofenadine on stimulated arachidonic acid metabolism. Human monocytes, isolated from blood and donated by 5 healthy volunteers, were either incubated for 20 h with 10 microg/ml lipopolysaccharide, with and without fexofenadine (10(-8)-10( 3) mol/l, n = 8-19), or were incubated for 20 h, with and without fexofenadine, and then stimulated with 0.5 mg/ml zymosan for 2 h. Leukotriene B4 (LTB4), LTC4, LTD4 and LTE4, prostaglandin E2 (PGE2) and F2alpha (PGF2alpha) production was determined by enzyme immunoassay. Zymosan-stimulated production of LTC4, LTD4 and LTE4 was significantly inhibited by clinically relevant concentrations of fexofenadine HCl: 10(-7) mol/l (22% inhibition vs. control, p = 0.008) and 10(-6) mol/l (24% inhibition vs. control, p = 0.020). Higher concentrations of fexofenadine (10(-4) and 10(-3) mol/l) inhibited LTB(4) generation. Lipopolysaccharide-stimulated production of PGE2 was significantly inhibited by fexofenadine HCl 10(-6) mol/l (26% inhibition, p = 0.035) and 10(-5) mol/l (40% inhibition, p = 0.001). Higher concentrations of fexofenadine HCl (10(-4) and 10( 3) mol/l) significantly inhibited PGF2alpha production by 50% (p = 0.026) and 63% (p = 0.001), respectively. Fexofenadine, at both clinically relevant and higher concentrations, inhibits LTC4, LTD4, LTE4 and PGE2 in cultured human monocytes. These additional anti-inflammatory properties may underlie the relief of nasal congestion observed in clinical studies. PMID- 16254457 TI - Tumor markers in breast cancer- European Group on Tumor Markers recommendations. AB - Recommendations are presented for the routine clinical use of serum and tissue based markers in the diagnosis and management of patients with breast cancer. Their low sensitivity and specificity preclude the use of serum markers such as the MUC-1 mucin glycoproteins (CA 15.3, BR 27.29) and carcinoembryonic antigen in the diagnosis of early breast cancer. However, serial measurement of these markers can result in the early detection of recurrent disease as well as indicate the efficacy of therapy. Of the tissue-based markers, measurement of estrogen and progesterone receptors is mandatory in the selection of patients for treatment with hormone therapy, while HER-2 is essential in selecting patients with advanced breast cancer for treatment with Herceptin (trastuzumab). Urokinase plasminogen activator and plasminogen activator inhibitor 1 are recently validated prognostic markers for lymph node-negative breast cancer patients and thus may be of value in selecting node-negative patients that do not require adjuvant chemotherapy. PMID- 16254458 TI - The Fem1a gene is downregulated in Rhabdomyosarcoma. AB - Rhabdomyosarcoma (RMS) is the most common soft tissue neoplasm of children, and those metastatic at presentation have a poor prognosis. RMS development is related to defective skeletal muscle differentiation, involving a variety of cell signaling and transcriptional control pathways, including aberrant hedgehog signaling. Here we evaluate Fem1a, a gene highly expressed in skeletal muscle, as a candidate for involvement in RMS. Fem1a is a homolog of fem-1, which controls cell fate decisions in the sex determination pathway of Caenorhabditis elegans, a pathway with homology to mammalian hedgehog signaling. We show that Fem1a expression is activated during myocyte differentiation of C2C12 myoblasts, and this expression is largely confined to the terminally differentiating pool, not to the satellite-cell-like quiescent reserve cell pool. We find that the human homolog, FEM1A, is downregulated in all of 8 different human RMS cell lines, including those derived from embryonal and alveolar RMS. Using mouse genetic models of RMS development, we further show that Fem1a is consistently downregulated in primary RMS from Ptch1+/- mice, from p53-/- mice, from p53+/-; Ptch1+/- mice, and from HGF/SF-Ink4a/Arf-/- mice. Therefore, Fem1a downregulation may be involved in, and/or a marker of, an early cell fate defect fundamental to RMS pathogenesis. PMID- 16254459 TI - Effect of promoter methylation of the p16 gene on phosphorylation of retinoblastoma gene product and growth of hepatocellular carcinoma cells. AB - The biological significance of hypermethylation of p16 gene promoter in human hepatocellular carcinoma (HCC) cells remains to resolved. In order to clarify the significance of methylation of p16 gene promoter, we examined the methylation status of p16 gene in association with phosphorylation of retinoblastoma gene product (pRb) and cell growth in human HCC cell lines. The presence of methylation was examined by methylation-specific PCR. Expression and phosphorylation of p16 and pRb were examined by Western blot analysis. Genetic changes were analyzed by multiplex PCR and DNA sequencing. The effect of demethylation of p16 was assessed by cell growth. p16 gene promoter was methylated in HuH7 and HLF cells. The demethylating agent, 5-aza-2-deoxycytidine (5-Aza-CdR), upregulated p16 mRNA in HuH6 and HuH7 cells. 5-Aza-CdR increased p16 protein expression in HuH6, HuH7, and HLF cells, and it clearly decreased the phosphorylation level of pRb in HuH6, HuH7 and PLC/PRF/5 cells. Treatment with 5 Aza-CdR inhibited the growth of HuH7 cells. Homozygous deletion and significant mutations were absent. Methylation in the p16 promoter region is biologically significant, being associated with phosphorylation of pRb and cell growth in human HCC cells. PMID- 16254460 TI - Chromosomal abnormalities associated with neck nodal metastasis in nasopharyngeal carcinoma. AB - Neck lymphatic metastasis represents the single most important clinical prognostic factor in nasopharyngeal carcinoma (NPC), but underlying genetic mechanisms remain ill defined. In this study 23 samples of primary tumor (PT) and 9 of neck lymph node metastasis (NLNM) obtained from NPC patients were analyzed by comparative genomic hybridization (CGH) coupled with tissue microdissection and degenerate oligonucleotide primer-polymerase chain reaction (DOP-PCR). A similar pattern of chromosomal abnormalities was seen in PT and NLNM, the common aberrations were gains on 5p, 12p, 12q and 18p and deletions on 1p, 3p, 9q, 14q, 17p and 16q. However, NLNMs, but not PTs, also exhibited frequent losses on 9p, 16p, 17q, 20q, 21p, 21q and 22q and gains on 8q and 8p. The most frequent unique aberration in NLNMs was loss on 16p, observed in 100% (9/9) NLNMs tested, as well as loss of 20q, observed in 77.8% of tumors tested. For the first time, we report that a gain on 8p and a loss at 20q is common to NLNMs. The analysis furthermore suggests that specific alterations, e.g. losses of 9p, 16p, 7q, 20q, 21p, 21q, 22q and gains on 8q and 8p are associated with NLNM of NPC, and that these alterations may be involved in the onset and/or progression of a metastatic phenotype. PMID- 16254461 TI - Cellular retinoic acid binding protein I: expression and functional influence in renal cell carcinoma. AB - Despite the known anti-proliferative and tumor-suppressive effects seen with retinoic acid (RA), treatment of metastatic renal cell carcinoma (RCC) failed to meet the initial expectations. As the exact mechanisms of action of RA and especially the role of the cellular RA binding proteins (CRABP) have not been elucidated yet, we investigated the expression of CRABP-I and its potential influence on RA response in RCC. Real-time RT-PCR analysis disclosed a significant lack of CRABP-I expression in four RCC cell lines and 12 primary RCC samples; in contrast, high expression levels were found in the respective adjacent normal kidney tissue. To further investigate the impact of CRABP-I on RA response in RCC, A-498 RCC cells were employed as a cellular model system. CRABP I was stably transfected into A-498 cells which consequently displayed substantial resistance to all-trans (ATRA) and 9-cis RA compared to vector controls lacking CRABP-I. Comparison of gene expression profiles of ATRA-treated CRABP-I-expressing A-498 cells and vector controls revealed specific regulation of 54 of approximately 20,000 genes tested on a selected human CodeLink UniSet Bioarray, with a prominent modulation of genes involved in transcriptional control, signaling, apoptosis, cell cycle regulation and metabolism. The genetic changes reported here contribute to a better understanding of the role of RA in RCC. They also provide new insights into CRABP-I-mediated signaling and gene expression. PMID- 16254462 TI - Downregulation of human kallikrein 10 (KLK10/NES1) by CpG island hypermethylation in breast, ovarian and prostate cancers. AB - OBJECTIVE: The human kallikrein 10 (KLK10)/normal epithelial cell-specific-1 (NES1) gene is highly expressed in normal mammary, ovary and prostate cells, but its expression is dramatically decreased in cancer cell lines. Recently, it has been shown that CpG island hypermethylation of the KLK10 gene is responsible for the tumor-specific loss of KLK10 gene expression in certain breast cancer cell lines. METHOD: We examined the role of CpG island hypermethylation in the tumor specific loss of KLK10 expression in breast, ovarian and prostate cancers. We treated cells with the demethylating agent 5-aza-2'-deoxycytidine (dC) and monitored changes in KLK10 mRNA by RT-PCR and secreted hK10 protein expression by ELISA. The following cell lines were used: MDA-MB-231, MDA-MB-468, MCF-7, ZR-75 1, T-47D and BT-474 (breast); BG-1, MDAH-2774, HTB-75, HTB-161, PA-1 and ES-2 (ovary), and LNCaP and PC-3 (prostate). RESULTS: Upregulation of KLK10 mRNA levels, which was accompanied by an increase in secreted hK10 protein concentration, was observed for a subset of breast, ovarian, and prostate tumor cell lines after 5-aza-2'-dC. Genomic sequencing of sodium-bisulfite-treated DNA demonstrated that CpG sites within the KLK10 gene exon 3 were highly methylated. Hypermethylation of exon 3 CpG regions was also detected in primary ovarian cancers. CONCLUSION: These data suggest that CpG island hypermethylation plays an important role in the downregulation of kallikrein 10 mRNA and protein expression, but it cannot explain the pattern of expression of this gene in all cell lines or tissue tested. PMID- 16254464 TI - Chemomechanical caries removal for children. AB - Pain is still an actual problem in dentistry, and a usage of anaesthetics can be still limited, so new ways of caries treatment are still being searched in order to ease this process both to a patient and a doctor. Efficiency of chemomechanical tooth treatment to children was studied. 30 children within two age groups of 3-6 and 7-13 years took part in this research, and their teeth with caries were treated in two different ways--chemomechanical and traditional. Teeth with the same class of cavities were treated. There are presented records about the need of anaesthetics' usage, cleaning duration and patients' complaints in this study. Results show that chemomechanical treatment could be as effective as traditional one, causes less pain and lowers the need for anaesthetics. There was noticed less removal of solid tissue cleaning carious cavity. PMID- 16254463 TI - Comparative analysis of the effectiveness of the mandibular angle fracture treatment methods. AB - 425 patients with mandibular angle fractures were treated at the Hospital of Kaunas University of Medicine (HKUM) Clinic of Maxillo-Facial Surgery. Treatment included the application of closed fracture fragments fixation methods (wire splint fixation, and Kirschner wire osteosynthesis), and methods of open fixation -osteosynthesis using the supra-periosteal miniplate, and osteosynthesis using supra-osseous Zes Pol plate (the latter method was modified by the authors). Relative computerized densitometry showed that closed fixation methods result in a faster healing of fractures. The findings of the pain threshold testing showed that open fixation methods more severely damage the function of the lower alveolar nerve. Using closed fixation methods, osteomyelitis occurred in 5.3% of cases, while using open fixation methods--in 15.3% of cases. Thus, the authors of the article maintain that when mandibular angle fractures, in the presence of suitable conditions, closed fracture fragments fixation methods should be given a priority. PMID- 16254465 TI - The microbiological status of patients with periodontitis in southern Estonia after non-surgical periodontal therapy. AB - Our objective was to investigate the presence of periodontal pathogens in comparison with the total degree of microorganisms after non-surgical periodontal therapy. The study material consisted of microbiological samples from periodontal pockets originating from 140 consecutive patients with chronic generalized severe periodontitis. The subgingival samples from periodontal pockets were obtained by a sterile curette, placed into 2 ml of the VMGA III medium, homogenized and serially diluted in the Brucella broth. 100 microl aliquots from the dilutions were inoculated onto the Brucella and the TSBV agar. The plates were incubated in an anaerobic chamber and under microaerobic conditions. The isolates were identified according to colonial and cellular morphology, the potency disk pattern, and the biochemical profiles. After instrumentation, no periodontal pathogens were isolated in 46 (33%) patients, while 94 patients (67%) were infected with one to five different periodontal pathogens. However, higher degree of the total microflora was positively correlated with number of isolated pathogens, a putative indicator of their presence. Therefore, due to the occurrence of residual microorganisms after non-surgical mechanical treatment, information about the pattern of these pathogens is needed for application of antimicrobial therapy. The level of microbial load in gingival pockets, including both pathogenic and non-pathogenic species, is one of the determinants of presence of residual pathogens after non-surgical periodontal therapy. PMID- 16254467 TI - Effectiveness evaluation of different suction systems. AB - Microorganisms of the patient's oral cavity and his/her blood and saliva may cause different air-borne and blood-borne infectious diseases among odontologists and their assistants who work with patients. Quantitative analysis and spatial distribution analysis of the environmental spread of oral liquid and cooling liquid mixture were performed during this study. Effectiveness of suction systems of four types was evaluated: without suction, using a small-size suction pump alone, using a small-size and large-size suction pumps, using a small-size suction pump together with an experimental extra-oral aspirator. Quantitative changes of the water aerosol, which enters the environment during the preparation of teeth, were determined in respect of the used suction systems. The small-size pump system together with an experimental extra-oral suction system eliminated best the aerosol formed during the preparation. PMID- 16254466 TI - The response of peripheral blood neutrophils unstimulated and stimulated by prodigiosan to smoking. AB - Alterations of neutrophils function by tobacco products may play a central role in the pathogenesis of periodontal diseases and several smoking related systemic diseases. The aim of the study was to assess a response of peripheral blood neutrophils unstimulated and stimulated by prodigiosan to smoking. MATERIALS AND METHODS: The study included 17 smoking men that addressed for treatment of various odontological problems to out patient department of Kaunas University Clinic. The subjects were 22-43 years of age and systemic diseases free. All subjects answered to questions about smoking habits. To assess the response of neutrophils (unstimulated and stimulated by prodigiosan) to smoking the chemiluminescence and nitroblue tetrazolium test were used to measure their oxidative metabolism. RESULTS: After smoking both neutrophils unstimulated and stimulated with prodigiosan showed considerable extracellular chemiluminescence responses, but the first one reached maximal value in 45 min, while the second one in 15 min after smoking. There was no total (intra and extracellular) chemiluminescence response of neutrophils unstimulated and stimulated by prodigiosan to smoking. Neutrophils exposed to smoking showed a significant increase in their nitroblue tetrazolium reduction. CONCLUSION: Both stimulated by prodigiosan and unstimulated peripheral blood neutrophils release reactive oxygen species extracellulary which may alter the pathogenic processes in periodontal and other systemic diseases. PMID- 16254468 TI - Dietary and oral hygiene habits in children with type I diabetes mellitus related to dental caries. AB - The aims of the study were to evaluate differences in dietary, oral hygiene habits and social class in children with Type I diabetes mellitus (DM), compared to non-diabetics, and to investigate relationship between selected caries-risk factors and caries experience in diabetics. MATERIAL AND METHODS: 70 children with Type I DM and 70 age- and sex-matched non-diabetic controls were included in the study. Metabolic control of diabetes was categorized into well- to- moderately-controlled and poorly-controlled groups based on glycosylated haemoglobin HbA1c. The study was based on the data obtained from the questionnaire including information about dietary and oral hygiene habits, pattern of dental visits and social class. Results showed that the diabetic children had more frequent main meals and less snacking than their controls: the mean number of main meals/day was 4.33 (SD = 0.93) in the diabetics, and 2.53 (SD = 0.85) in the controls. Significantly less diabetics (43%) used sweet drinks than their controls (79%). There were no differences according to the frequency of toothbrushing as well as frequency of dental visits between the diabetics and controls, however, significantly more diabetics reported that they never used dental floss than non-diabetics. There were no significant differences in the diet, toothbrushing frequency between the diabetics with different metabolic control. Multiple logistic regression analysis showed that among caries risk associated variables only age of children (OR = 1.98; CI = 1.23-3.19) and level of metabolic control of diabetes (OR = 4.65; CI = 1.28-16.89) were statistically significantly associated with high caries experience in the diabetics. CONCLUSIONS: Frequent consumption of sweet drinks and snacks can influence caries development in children. Amongst the diabetics, the differences in caries prevalence can be explained by combination of biological and behavioral factors rather than single dietary or oral hygiene elements. PMID- 16254469 TI - Halitosis manifestation and prevention means for patients with fixed teeth dentures. AB - The objective of this research is to analyse the causal relationship between construction of fixed bridge dentures and the intensity of halitosis manifestations, as well as to establish basic hygiene requirements for construction of fixed dentures which would completely exclude retention of food particles and avoid bad breath. 48 patients (36 men and 12 women), who use fixed dentures for 2-10 years, have been involved in this research. 26 patients wore fixed bridge dentures made of punched tooth crowns, the other 22 patients wore cast fixed dentures. The obtained measurements of halitosis magnitude point to the close connection between bad breath and the construction of fixed dentures. Fixed dentures with tooth crown laps, saddle intermediate parts, as well as denture constructions, which impede complex of mouth hygiene measures, cause bad breath. In this research, the condition of patients' teeth, periodontium, and oral cavity hygiene have been evaluated as satisfactory; the tongue is not perceptibly coated, and patients etiologically have not experienced problems caused by respiratorial or gastrointestinal diseases. The examined patients have not complained of xerostomia problems. In conclusion, it should be admitted that fixed dentures, which make difficult or even completely impede the complex of oral cavity hygiene measures, intensify the development of halitosis. PMID- 16254470 TI - Clinical effectiveness of the Twin block appliance in the treatment of Class II Division 1 malocclusion. AB - The aim of the present study was to assess clinical effectiveness of Class II Division 1 malocclusion treatment with Twin block appliance. MATERIAL AND METHODS: Analysis of cephalometric radiographs of 34 Class II Division 1 patients treated with Twin block appliance was performed before and after treatment. A control group was generated from the normative growth data published by Bhatia and Leighton. The treatment effect was calculated by subtracting the natural growth change from the treatment change. This was then compared to twice the method error to see if the treatment change was clinically significant. RESULTS: Mean mandibular length as measured from point Art to point Pog increased by 6.4 mm in the Twin-block group compare with 4.1 mm in the control group. The overjet during treatment was reduced by 4.9 mm. Relative to the maxilla upper incisor tipped backward by 6.7 degrees and in the control group natural growth proclined them by 2.4 degrees. Lower incisor after the treatment tipped forward and the angle between long axis of lower incisor and mandibular plane increased by 3.3 degrees, whereas in the control group they stay almost in the same position, proclination only 0.7 degrees. CONCLUSIONS: Twin block appliance clinically significantly increases mandibular length (net effect 2.3 mm) and reduce overjet (net effect 4.9 mm). Modification of the Twin block appliance by acrylic extension to cover the edges of lower incisors reduce dentoalveolar tipping and maximize skeletal changes. PMID- 16254471 TI - Dental treatment needs in Lithuanian adolescents. AB - The aims were to estimate dental treatment needs of 15-16-year-olds in Lithuania and to relate a number of background factors such as gender, urbanization and residency to possible differences in dental treatment needs. A total of 885 adolescents in 22 randomly pre-selected areas were clinically examined. The following criteria of dental treatment needs were used: tooth with primary caries, secondary caries, tooth which needs a non-operative treatment, tooth with trauma, which needs a restorative treatment, and tooth which needs either an endodontic treatment, a crown or an extraction. Only 7.7 % of all participants had all their teeth sound. The treatment due to primary caries was the most required dental treatment modality among Lithuanian adolescents. Significantly fewer teeth required treatment due to other aforementioned reasons. Significant differences in most dental treatment needs between boys and girls were found, whereas in relation to urbanization and area of residency the differences were less pronounced. PMID- 16254472 TI - Mandibular pubertal growth spurt prediction. Part one: Method based on the hand wrist radiographs. AB - Many orthodontic treatment modalities will yield a better result in less time if properly correlated with the unique facial growth patterns of the patients. The pubertal growth spurt depends on gender and varies in relationship to the chronologic age. General skeletal maturity usually is used as an indicator to predict timing of mandibular growth velocity peak. Hand-wrist radiographic evaluation is one of the diagnostic tools currently available to determine whether the pubertal growth has started, is occurring or has finished. The overview of topic related literature and skeletal maturity assessment (SMA) system developed by L. Fishman are presented. The SMA system is based on eleven discrete adolescence skeletal maturational indicators of hand-wrist bones, covering the entire period of adolescent development. Maturational stage and level demonstrated close correlation with maxillary and mandibular growth velocity, amount of incremental growth and timing. Clinical indications for the use of hand-wrist radiographs to assess skeletal maturity are provided. PMID- 16254473 TI - Factors influencing the removal of posts. AB - Root canal retreatment in teeth restored with intraradicular posts has attracted interest due to the difficulties of their removal without weakening, perforating or fracturing the remaining root structure. The use of ultrasonic devices has been suggested by some authors to facilitate post removal, reducing the possibility of fractures or root perforations. The purpose of this study was to determine the efficacy of cast posts removal by ultrasonic device regarding post length, adaptation of post to root canal walls and cement type. MATERIAL AND METHODS: Seventy-seven single rooted endodontically treated teeth restored with cast posts were included to this study. Post removal was processed with ultrasonic and time for this was recorded. The length and width of the post, quality of post adaptation, type of cement were evaluated according to radiographic findings and clinical records. RESULTS: Mean value of time needed for post removal was 14.15 (SD +/- 8.57) minutes. A strong correlation was observed between the time of post removal and post length (r=0.620, p=0.000). The mean time required for the removal of posts cemented with zinc phosphate was 11.36 min (SD +/- 5.84) and for the posts cemented with resin modified glass ionomer was 15.37 min (SD +/- 3.83). Statistically significant difference in the time needed for the posts cemented with different cements was observed (p = 0.002). The mean time needed for the post removal with the inappropriate adaptation was 10.1 min (SD +/- 6.0) and the time for the post with appropriate adaptation was 15.7 min (SD +/- 8.9). Statistically significant difference between these two groups was observed for the time of posts removal (p = 0.003). According to the results of linear regression model test, 50% of variation in time needed to remove posts was explained by following variables: post length, post adaptation and the cement type. The total regression model was highly significant (p = 0.000). CONCLUSIONS: The time taken for post removal depended on post length, its adaptation in the root canal and on the type of luting cement. Root fractures are unlikely to occur with good case selection. PMID- 16254474 TI - Oral hygiene in children with type I diabetes mellitus. AB - Oral hygiene is an important etiological factor related to oral health status in children. The aim of the study was to evaluate the oral hygiene status in children with type I diabetes mellitus (DM) and in their non-diabetic controls, and to correlate it with the health condition of the gingival tissues. MATERIALS AND METHODS: Seventy 10-15 year-old children (mean age 13.6, SD=1.6) with type I DM and 70 their age and sex-matched non-diabetic controls were included in the study. The metabolic control of DM was categorized into well- to- moderately controlled and poorly-controlled diabetes groups based on glycosylated haemoglobin HbA1c. The oral hygiene and gingival status were assessed using the Simplified Oral hygiene index OHI-S (Greene-Vermillion) and gingival index GI (Loe-Silness), respectively. Student's t, Mann-Whitney U or chi-square tests and linear regression were used in the statistical analyses. RESULTS: There were no statistically significant differences in the mean values of OHI-S between the diabetics and non-diabetics, however the mean calculus index (CI-S) was significantly higher and the mean plaque index (DI-S) was significantly lower in diabetic subjects than non-diabetics (p<0.05). The children with type I DM had significantly higher mean values of GI compared with the non-diabetic subjects (0.15, SD=0.37 and 0.05, SD=0.19 respectively) (p<0.05). Healthy gingiva was recorded in 73% diabetics, and 87% of the non-diabetics (p<0.05). The poorly controlled diabetics had higher mean gingival, plaque, calculus indices than well moderately controlled subjects, although the differences were not statistically significant. Statistically significantly higher mean OHI-S index was found in children with poor control of diabetes compared with the well-moderately controlled diabetics (1.39, SD=0.75 and 1.07, SD=0.53, respectively) (p<0.05). The level of gingivitis statistically significantly correlated with the levels of calculus in both study groups (p<0.001). No difference in toothbrushing habits was revealed between the study subjects. CONCLUSION: Despite similar oral hygiene habits, the children with type I DM were more prone to calculus accumulation which seemed to be a predisposing factor in development of gingivitis in these individuals. PMID- 16254475 TI - Critical assessment of temporomandibular joint clicking in diagnosing anterior disc displacement. AB - OBJECTIVES: The objective of this study was to compare temporomandibular joint (TMJ) findings from clinical examination and magnetic resonance imaging (MRI) for diagnosing anterior disc displacement. MATERIAL AND METHODS: 114 subjects with at least one sign of temporomandibular disorders (tenderness/pain on palpation of the joints or muscles, TMJ sounds, pain or deviation during maximum mouth opening (active/passive)) and 193 controls underwent MRI after proper history taking and assessment of clinical symptoms. RESULTS: 464 joints with no disk displacement (NDD), 114 joints with reducible displaced disk (RDD), and 36 joints with permanently displaced disk (PDD) were confirmed on magnetic resonance imaging (MRI). CONCLUSION: The results suggest that anterior disc displacement of the TMJ can not be diagnosed with considerable accuracy through the use of clinical examination only. PMID- 16254477 TI - Understanding the drug development process. AB - In the United States, new drug development is a complex, highly regulated process that often involves academic institutions, the pharmaceutical industry, and government agencies. Much of the primary and clinical research conducted in the United States is funded by the National Institutes of Health. The Food and Drug Administration (FDA) provides regulatory review and oversight of the drug development process. The Investigational New Drug application is the primary mechanism that the FDA uses to regulate clinical testing in humans. A New Drug Application summarizing all of the data necessary to perform risk/benefit analysis of the new drug is submitted to the FDA for review and approval once sufficient testing has been completed. Recent research into the pathogenesis of inflammatory bowel disease has yielded numerous drug targets resulting in a multitude of molecules and biologic agents under varying stages of development and a consequent need for clinical testing. A thorough understanding of the regulatory development process and the National Institutes of Health funding process is necessary for any investigator involved in clinical trials. PMID- 16254478 TI - Ethical issues, safety, and data integrity in clinical trials. AB - Published in 1974, the Belmont Report established the ethical principles for conducting clinical research in the United States. The essential concepts are respect for the research participant, beneficence for society at large, and justice (equal access to participation and equal treatment) toward subjects in a research study. These principles are applied through the use of informed consent, risk/benefit assessment, and the impartial selection of study subjects. Strict adherence to these criteria often results in conflicts of interest, which the investigator must anticipate and manage. Investigators must also be thoroughly acquainted with the principles of Good Clinical Practice and regulatory requirements. Recent implementation of the Privacy Rule now requires the investigator to protect not only the safety but the privacy of the research subject. While the regulatory obligations can appear onerous, strict compliance results in clinical research that is safe, scientifically sound, and ethical. PMID- 16254479 TI - Design issues and outcomes in IBD clinical trials. AB - Successful clinical trials in inflammatory bowel disease are based on precise definitions of study populations, standardized and well-defined interventions, reliable indices of disease activity, and clearly stipulated outcome measures. Interpretation of research results is often complicated by the differentiation of goals of therapy (remission induction and maintenance, quality of life) and the multitude of patient subsets. Choosing the correct patient subtype to enroll in a clinical trial is critical to the results of a study, its conclusions, and its applicability to the clinical setting. Validated, easy-to-use disease activity indices allow interpretation of results across trials. The use of biomarkers as surrogate clinical endpoints provides reproducibility, ease of statistical handling as a continuous variable, and consistent measurement of response to treatment. Despite these potential advantages, biomarkers of disease activity lack specificity and will need to be validated against clinically meaningful outcomes. Careful subject selection, standardization of disease activity indices, and precise outcome measurement are the keys to continued improvement of the inflammatory bowel disease research process. PMID- 16254480 TI - Site organization and management. AB - The principal investigator is responsible for everything involved in the conduct of a clinical research study. Prior to the initiation of any clinical trial, an investigator must become acquainted with the material requirements, personnel needs, and best practices involved in the conduct of the trial. Commitment to a clinical trial should not be taken lightly because even a simple study may require a major investment of staff, space, and time. Standard operating procedures help to standardize staff training and improve regulatory compliance. Reasons for participation in clinical research may differ between community and academic gastroenterologists, but responsibility for patient care, regulatory requirements, and assumption of accountability are identical. Careful attention to the details of site organization, administrative requirements, and patient recruitment and retention all contribute to the successful performance of clinical research. PMID- 16254481 TI - State of the art: IBD therapy and clinical trials in IBD. AB - Inflammatory bowel diseases (IBD) encompass Crohn's disease and ulcerative colitis, which are diseases characterized by chronic intestinal inflammation. IBD is believed to result from predisposing genetic and environmental factors (specific antigens and pathogen-associated molecular patterns) acting on the immunoregulatory system and causing inflammation of the gastrointestinal mucosa. IBD may be the result of an imbalance of effector (proinflammatory) and regulatory T-cell responses. Three scenarios indicative of the outcome of this balance exist in animal models: balanced effector and regulatory T cells resulting in a normal controlled inflammation; overactive effector T cells resulting in inflammation and disease; and an absence of regulatory T cells resulting in uncontrolled inflammation and severe, aggressive disease. The number of products under study for the treatment of IBD has increased from 3 products and 1 target in 1993 to more than 30 products and more than 10 targets in 2005. The number of products under development and continued investigations into the pathogenesis of IBD emphasize the need to expand clinical research efforts in IBD. PMID- 16254487 TI - The neuropathology of autism: a review. AB - Presented is a review of recent progress in the understanding of autism based on investigations of donated human brain tissue. Autism is a pervasive developmental disorder by the American Psychiatric Association's Diagnostic and Statistical Manual of Mental Disorders (DSM-IV) criteria, manifesting by age 3 and characterized by impairments in social interaction and communication, as well as restricted, repetitive, stereotyped patterns of behavior. Based on reported neuropathologic findings, these characteristic behaviors are clinical manifestations of both pre- and postnatal alterations. This review summarizes the current data obtained from postmortem brain studies in the areas of stereology, neurotransmitter systems/synaptic processes, molecular mechanisms, and neuroimmunology. In addition, we discuss current research strategies designed to facilitate translational research and maximize the yield of precious resources (e.g. the Autism Tissue Program), highlight barriers to research, and consider future trends. PMID- 16254488 TI - Coordinated and spatial upregulation of arc in striatonigral neurons correlates with L-dopa-induced behavioral sensitization in dyskinetic rats. AB - Although oral administration of L-Dopa remains the best therapy for Parkinson disease, its long-term administration causes the appearance of abnormal involuntary movements such as dyskinesia. Although persistent striatal induction of some genes has already been associated with such pathologic profiles in hemiparkinsonian rats, molecular and cellular mechanisms underlying such long term adaptations remain to be elucidated. In this study, using a rat model of L Dopa-induced dyskinesia, we report that activity regulated cytoskeletal (Arc) associated protein is strongly upregulated in the lesioned striatum and that the extent of its induction further varies according to the occurrence or absence of locomotor sensitization. Moreover, Arc is preferentially induced, along with FosB, nur77, and homer-1a, in striatonigral neurons, which express mRNA encoding the precursor of dynorphin. Given the likely importance of Arc in the regulation of cytoskeleton during synaptic plasticity, its upregulation supports the hypothesis that a relationship exists between cytoskeletal modifications and the longlasting action of chronically administrated L-Dopa. PMID- 16254489 TI - Histology-based expression profiling yields novel prognostic markers in human glioblastoma. AB - Although the prognosis for patients with glioblastoma is poor, survival is variable, with some patients surviving longer than others. For this reason, there has been longstanding interest in the identification of prognostic markers for glioblastoma. We hypothesized that specific histologic features known to correlate with malignancy most likely express molecules that are directly related to the aggressive behavior of these tumors. We further hypothesized that such molecules could be used as biomarkers to predict behavior in a manner that might add prognostic power to sole histologic observation of the feature. We reasoned that perinecrotic tumor cell palisading, which denotes the most aggressive forms of malignant gliomas, would be a striking histologic feature on which to test this hypothesis. We therefore used laser capture microdissection and oligonucleotide arrays to detect molecules differentially expressed in perinecrotic palisades. A set of RNAs (including POFUT2, PTDSR, PLOD2, ATF5, and HK2) that were differentially expressed in 3 initially studied, microdissected glioblastomas also provided prognostic information in an independent set of 28 glioblastomas that did not all have perinecrotic palisades. On validation in a second, larger independent series, this approach could be applied to other human glioma types to derive tissue biomarkers that could offer ancillary prognostic and predictive information alongside standard histopathologic examination. PMID- 16254490 TI - Somatostatin receptor type 2 undergoes plastic changes in the human epileptic dentate gyrus. AB - Temporal lobe epilepsy (TLE) is characterized by hippocampal sclerosis together with profound losses and phenotypic changes of different classes of interneurons, including those expressing somatostatin (SRIF). To understand the functional significance of the plasticity of SRIF transmission in TLE, unraveling the status of SRIF receptors is, however, a prerequisite. To address this issue, we characterized expression and distribution of the major SRIF receptor, the sst2 subtype, in hippocampal tissue resected in patients with TLE using complementary neuroanatomic approaches. In patients with hippocampal sclerosis, the number of cells expressing sst2 receptor mRNA as well as sst2 receptor-binding sites and immunoreactivity decreased significantly in the CA1-3, reflecting neuronal loss. By contrast, in the dentate gyrus, sst2 receptor mRNA expression was strongly increased in the granule cell layer, and sst2 receptor-binding sites and immunoreactivity was preserved in the inner but decreased significantly in the outer molecular layer. In this latter region, pronounced changes in SRIF terminal fields were observed. Decreased receptor density in the distal dendrites of granule cells is likely to reflect downregulation of sst2 receptors in response to physiopathologic release of SRIF. Because sst2 receptors have anticonvulsant and antiepileptogenic properties, this phenomenon may contribute to the etiology of TLE seizures. PMID- 16254491 TI - Brain damage after heat stroke. AB - Cerebellar syndromes and radiologic cerebellar atrophy after hyperpyrexia have occasionally been reported, mostly in neuroleptic malignant syndromes, but neuropathologic studies are extremely rare. We studied 3 patients (a 74-year-old woman, a 63-year-old man, and an 80-year-old man) who had heat stroke during heat waves in France. One patient had generalized seizures and died 28 hours after admission. The other patients survived one month and 2 months after admission; both had palatal myoclonus, and in one case, magnetic resonance imaging showed high signal intensity in the cerebral peduncles. The main neuropathology in the 3 cases was severe diffuse loss of Purkinje cells associated with heat shock protein 70 expression by Bergmann glia. In situ end labeling was negative in surviving Purkinje cells, suggesting that the mechanism of neuronal death was not apoptosis. Degeneration of Purkinje cells axons resulted in myelin pallor of the white matter of the folia and of the hilum of the dentate nuclei. DNA internucleosomal breakages were identified by in situ end labeling in the dentate nuclei and centromedian nuclei of the thalamus and were associated with degeneration of the cerebellar efferent pathways: superior cerebellar peduncles, decussation of the superior cerebellar peduncles (Wernekinck commissure), and dentatothalamic tract. These findings suggest that the mechanisms of neuronal death in the dentate nuclei and centromedian nuclei of the thalamus was different from that in Purkinje cells and more likely resulted from deafferentation. Ammon's horn and other areas susceptible to hypoxia were spared. These observations confirm the selective vulnerability of Purkinje cells to heat induced injury and involvement of the cerebellar efferent pathways in palatal myoclonus. PMID- 16254492 TI - Structure and stability of internodal myelin in mouse models of hereditary neuropathy. AB - Peripheral neuropathies often result in abnormalities in the structure of internodal myelin, including changes in period and membrane packing, as observed by electron microscopy (EM). Mutations in the gene that encodes the major adhesive structural protein of internodal myelin in the peripheral nervous system of humans and mice--P0 glycoprotein--correlate with these defects. The mechanisms by which P0 mutations interfere with myelin packing and stability are not well understood and cannot be provided by EM studies that give static and qualitative information on fixed material. To gain insights into the pathogenesis of mutant P0, we used x-ray diffraction, which can detect more subtle and dynamic changes in native myelin, to investigate myelin structure in sciatic nerves from murine models of hereditary neuropathies. We used mice with disruption of one or both copies of the P0 gene (models of Charcot-Marie-Tooth-like neuropathy [CMT1B] or Dejerine-Sottas-like neuropathy) and mice with a CMT1B resulting from a transgene encoding P0 with an amino terminal myc-tag. To directly test the structural role of P0, we also examined a mouse that expresses P0 instead of proteolipid protein in central nervous system myelin. To link our findings on unfixed nerves with EM results, we analyzed x-ray patterns from unembedded, aldehyde-fixed nerves and from plastic-embedded nerves. From the x-ray patterns recorded from whole nerves, we assessed the amount of myelin and its quality (i.e. relative thickness and regularity). Among sciatic nerves having different levels of P0, we found that unfixed nerves and, to a lesser extent, fixed but unembedded nerves gave diffraction patterns of sufficient quality to distinguish periods, sometimes differing by a few Angstroms. Certain packing abnormalities were preserved qualitatively by aldehyde fixation, and the relative amount and structural integrity of myelin among nerves could be distinguished. Measurements from the same nerve over time showed that the amount of P0 affected myelin's stability against swelling, thus directly supporting the hypothesis that packing defects underlie instability in "live" or intact myelin. Our findings demonstrate that diffraction can provide a quantitative basis for understanding, at a molecular level, the membrane packing defects that occur in internodal myelin in demyelinating peripheral neuropathies. PMID- 16254493 TI - TLR and NOD2 ligands induce cell proliferation in the rat intact spinal cord. AB - We demonstrate, by 5-bromo-2'-deoxyuridine (BrdU) tracing, the effects of peripheral administration of toll-like receptor (TLR) and NOD2 ligands (stimulators of the innate immune system) on the proliferation of spinal cord cells. Bolus injection of phosphorothioate oligonucleotides containing CpG motifs had no prominent effects on spinal cord neural progenitor cell proliferation, whereas single intraperitoneal injection of polyinosine-polycytidylic acid (Poly I:C, TLR3 ligand), lipopolysaccharide (LPS, TLR4 ligand), R848 (TLR7/8 ligand), or N-acetylmuramyldipeptide (MDP, Nod2 ligand) temporarily increased the number of BrdU(+) cells in the spinal cord. For Poly I:C- or R848-treated groups, the density of BrdU cells reached maximal levels on days 2 to 3 postinjection and then rapidly declined to baseline levels. Only a few of the proliferating cells were of microglial origin, but BrdU(+)/nestin(+) cells were found, suggestive of a proliferation of local progenitor cells. In addition, stimulation of cell proliferation correlated with activation of the innate immune system, that is, microglial cells. Interestingly, activation and cell proliferation was inhibited by corticosteroid dexamethasone but not by indomethacin. These findings suggest an intricate interaction of phylogenetically ancient cellular processes of the innate immune system and regeneration. PMID- 16254494 TI - Prenatal ischemia and white matter damage in rats. AB - Ischemia/reperfusion injury to the developing brain is a major cause of neurologic abnormalities in preterm infants. To investigate the underlying mechanisms, we modified a previously described rat model of unilateral uterine artery ligation on the 17th embryonic day (E17). Growth retardation was taken as an index of in utero ischemia, and pups born with a birth weight more than 2 standard deviations below that of controls were compared with the same-litter, normal-growth control pups born from the nonligated horn. Prenatal ischemia probably associated with hypoxia and followed by reperfusion at birth induced white matter damage at a developmental stage corresponding to extreme prematurity in humans. On P0 (day of birth), growth-retarded pups exhibited lesions in the cingular white matter and internal capsule with increased counts of activated microglial cells for 2 weeks compared with controls. Astrogliosis was detected in the injured white matter. On P3, increased apoptotic cell death was seen in O4 positive preoligodendrocytes, which were abnormally scarce on P7. Defective myelination, as assessed by myelin-binding-protein labeling, was detected until adulthood. The diffuse white matter damage in growth-retarded rats replicated the main features of white matter damage in human preterm infants. PMID- 16254495 TI - Axotomy of sympathetic neurons activates the metalloproteinase-2 enzymatic pathway. AB - We have previously shown that intraganglionic synapse disassembly consequent on superior cervical ganglion (SCG) neuron axotomy was preceded by the loss of the dystroglycan beta subunit (beta-DG) localized at the postsynaptic specializations. Because DG, a transmembrane molecular complex bridging the extracellular matrix to the cortical cytoskeleton, could be a physiological target of metalloproteinases (MMPs) 2 and 9, we investigated their possible involvement in the injury-induced intraganglionic synapse disassembly. In rat SCG, only MMP-2 was present and localized in both neurons and nonneuronal cells. After ganglion neuron axotomy, both MMP-2 activity and protein level increased, whereas the level of its mRNA was unchanged, suggesting prominent MMP-2 posttranslational regulation. mRNA and protein levels of the enzymes involved in the MMP-2 activation pathway, the membrane-type 1-MMP (MT1-MMP), and the tissue inhibitor of metalloproteinase-2 (TIMP-2) also increased after injury with a time course that correlated with that of MMP-2 activation. In addition, postganglionic nerve crush induced an increase in the beta-DG 30-kDa fragment produced by the MMP-dependent degradation of DG. These data suggest that MMP-2 activated during SCG neuron reaction to axotomy may degrade postsynaptic DG, contributing to the disruption of the molecular bridge between pre- and postsynaptic elements and disassembly of the intraganglionic synapses. PMID- 16254496 TI - Serotonin transporter abnormality in the dorsal motor nucleus of the vagus in Rett syndrome: potential implications for clinical autonomic dysfunction. AB - Autonomic dysfunction is prevalent in girls with Rett syndrome, an X-chromosome linked disorder of mental retardation resulting from mutations in the gene encoding methyl-CpG-binding protein 2 (MeCP2). This gene plays a role in regulating neuronal activity-dependent gene expression, including brain-derived neurotrophic factor (BDNF), which is a potent serotonergic (5-HT) neuronal growth factor. We analyzed selected parameters of the 5-HT system of the medulla in autopsied patients with Rett syndrome because of the role of BDNF in 5-HT cell development and because 5-HT plays a key role in modulating autonomic control. 5 HT neurons were identified by immunostaining for tryptophan hydroxylase, the biosynthetic enzyme for 5-HT. We quantitated the number of 5-HT cells in the medulla at 2 standardized levels in 11 Rett and 7 control cases. There was no significant difference in 5-HT cell number between the groups. We analyzed binding to the serotonin transporter (SERT) using the radioligand [(125)I]-RTI-55 with tissue autoradiography in 7 Rett and 5 controls in 9 cardiorespiratory related nuclei. In the dorsal motor nucleus of the vagus (DMX) (preganglionic parasympathetic outflow), SERT binding for the control cases decreased significantly over time (p = 0.049) but did not change in the Rett cases (p = 0.513). Adjusting for age, binding between the Rett and control cases differed significantly in this nucleus (p = 0.022). There was a marginally significant age versus diagnosis interaction (p = 0.06). Thus, altered 5-HT innervation and/or uptake in the DMX may contribute to abnormal 5-HT modulation of this major autonomic nucleus in patients with Rett syndrome. These data suggest hypotheses concerning 5-HT modulation of vagal function for testing in MeCP2 knockout mice to understand mechanisms underlying autonomic dysfunction in patients with Rett syndrome. PMID- 16254497 TI - 1. Guidelines on Paediatric Parenteral Nutrition of the European Society of Paediatric Gastroenterology, Hepatology and Nutrition (ESPGHAN) and the European Society for Clinical Nutrition and Metabolism (ESPEN), Supported by the European Society of Paediatric Research (ESPR). PMID- 16254510 TI - Gluten-free food labeling in the United States. PMID- 16254511 TI - Coping with ignorance: exploring pharmacologic management for pediatric functional abdominal pain. PMID- 16254512 TI - Ghrelin, energy balance, and weight in infants: the chicken or the egg? PMID- 16254513 TI - The composition of infant formula: a worldwide approach. PMID- 16254514 TI - Infant formulae: from ESPGAN recommendations towards ESPGHAN-coordinated global standards. PMID- 16254515 TI - Global standard for the composition of infant formula: recommendations of an ESPGHAN coordinated international expert group. AB - The Codex Alimentarius Commission of the Food and Agriculture Organization of the United Nations (FAO) and the World Health Organization (WHO) develops food standards, guidelines and related texts for protecting consumer health and ensuring fair trade practices globally. The major part of the world's population lives in more than 160 countries that are members of the Codex Alimentarius. The Codex Standard on Infant Formula was adopted in 1981 based on scientific knowledge available in the 1970s and is currently being revised. As part of this process, the Codex Committee on Nutrition and Foods for Special Dietary Uses asked the ESPGHAN Committee on Nutrition to initiate a consultation process with the international scientific community to provide a proposal on nutrient levels in infant formulae, based on scientific analysis and taking into account existing scientific reports on the subject. ESPGHAN accepted the request and, in collaboration with its sister societies in the Federation of International Societies on Pediatric Gastroenterology, Hepatology and Nutrition, invited highly qualified experts in the area of infant nutrition to form an International Expert Group (IEG) to review the issues raised. The group arrived at recommendations on the compositional requirements for a global infant formula standard which are reported here. PMID- 16254516 TI - Colonic chicken-skin mucosa in children with polyps is not a preneoplastic lesion. AB - Colonic polyps are common both in adults and children; however, the malignant potential varies according to the type of polyp. Most childhood polyps are solitary juvenile polyps, which have negligible malignant potential. Chicken-skin mucosa (CSM) is an endoscopic finding initially described associated with adenomatous polyps and adenocarcinoma, suggesting a preneoplastic lesion. Subsequently, CSM was described in association with juvenile polyps, suggesting that this mucosal finding is not a precursor to dysplasia. To determine whether CSM represents a preneoplastic lesion, we studied endoscopic colonic mucosal biopsies for markers of cell replication (Ki-67) and malignant transformation (p53) in mucosal biopsies of CSM, normal colonic tissue, tubular adenomas, and adenocarcinomas. Samples were subjected to immunostaining for the presence of Ki 67 and p53. The degree of Ki-67-positive staining cells was similar for CSM and normal colonic tissue, whereas there was significantly increased staining for both tubular adenomas and adenocarcinomas. There was no evidence of p53 staining in CSM and normal colonic mucosa, whereas there was varying degrees of staining in tubular adenomas and adenocarcinomas. The association of CSM with benign juvenile polyps and the absence of histologic markers for increased replication and malignant transformation support the notion that this endoscopic finding is not preneoplastic. Rather, CSM arises in proximity to polyps of all histologic types because of local mucosal damage. PMID- 16254518 TI - Clinical features of pathologic childhood aerophagia: early recognition and essential diagnostic criteria. AB - OBJECTIVE: This study investigated the early recognition and diagnosis of pathologic childhood aerophagia to avoid unnecessary diagnostic approaches and serious complications. METHODS: Between 1995 and 2003, data from 42 consecutive patients with pathologic childhood aerophagia, aged 2 to 16 years, were reviewed. An esophageal air sign was defined as an abnormal air shadow on the proximal esophagus adjacent to the trachea on a full-inflated chest radiograph. RESULTS: Of the 42 patients, the chief complaints were abdominal distention (52.4%), recurrent abdominal pain syndrome (21.4%), chronic diarrhea (11.9%), acute abdominal pain (7.1%) and others (7.2%). Mean symptom duration before diagnosis was 10.6 months (range, 1 to 60 months), and it exceeded 12 months for 16 (38.1%) patients. The clinical features common to all patients were abdominal distention that increased progressively during the day, increased flatus on sleep, increased bowel sound on auscultation and an air-distended stomach with increased gas in the small and large bowel by radiography. Visible or audible air swallowing (26.2%) and repetitive belching (9.5%) were also noted. Esophageal air sign was observed in 76.2% of the patients and in 9.7% of the controls (P=0.0001). The subgroups of pathologic childhood aerophagia divided by underlying associations were pathologic childhood aerophagia without severe mental retardation (76.2%), which consisted of psychological stresses and uncertain condition, and pathologic childhood aerophagia with severe mental retardation (23.8%). CONCLUSIONS: The common manifestations of pathologic childhood aerophagia may be its essential diagnostic criteria, and esophageal air sign may be useful for the early recognition of pathologic childhood aerophagia. Our observations show that the diagnostic clinical profiles suggested by Rome II criteria should be detailed and made clearer if they are to serve as diagnostic criteria for pathologic childhood aerophagia. PMID- 16254517 TI - Short-chain fatty acid induces intestinal mucosal injury in newborn rats and down regulates intestinal trefoil factor gene expression in vivo and in vitro. AB - BACKGROUND: Luminal administration of short-chain fatty acids (SCFAs) induces dose-dependent intestinal mucosal injury in newborn rats. However, the mechanism underlying the injurious effects of SCFAs on intestinal mucosa in neonates is unclear. Intestinal trefoil factor (ITF) is a factor important for the maintenance and repair of the intestinal mucosal barrier. Regulation of ITF gene expression by SCFAs may be involved as one of the mechanisms. OBJECTIVES: To examine the effect of butyrate-induced colonic injury on ITF gene expression in vivo and to determine the molecular mechanisms underlying the butyrate regulation of ITF gene expression in vitro. METHODS: Whole-section colonic tissues from 9- to 10-day-old Sprague-Dawley rats that have received butyric acid at two different concentrations (150 mmol/L and 300 mmol/L) and for different time periods were processed for total RNA extraction and Northern blot analysis. Littermates that received normal saline or lactic acid at 300 mmol/L served as controls. The effect of butyrate on ITF gene expression was also examined in vitro with human colonic epithelial LS 174T cells. To further define ITF gene regulation by butyrate, transient transfection assays were performed on a 930 bp human ITF promoter-luciferase reporter gene plasmid in LS174T cells with or without the presence of butyrate. RESULTS: Concurrent with mucosal injury, butyric acid inhibited ITF gene expression in colonic tissues of newborn rats as well as in intestinal epithelial cells in a dose- and time-dependent manner. Furthermore, butyrate reduced ITF promoter report gene activity in transfected LS174T cell, suggesting that butyric acid regulation of ITF gene is by way of a specific ITF promoter. CONCLUSIONS: Butyric acid induced-intestinal mucosal injury in newborn rats is associated with down-regulation of ITF gene expression. The changes in ITF gene expression in vivo may play a role in the pathogenesis of SCFA-induced intestinal mucosal injury. PMID- 16254519 TI - Does increased duration of exclusive breastfeeding protect against Helicobacter pylori Infection? The Newcastle Thousand Families Cohort Study at age 49-51 years. AB - OBJECTIVE: Helicobacter pylori acquired in childhood is an important risk factor for gastric carcinoma. Once colonization is established, infection may be carried for life. This study used prospectively recorded, detailed information on infant feeding and investigated the potential link between duration of exclusive breastfeeding in infancy and seropositivity at age 50 years, as measured by enzyme-linked immunosorbent assay. METHODS: H. pylori seropositivity at age 50 years was investigated among 407 individuals born in Newcastle in May and June 1947 and related to the duration of exclusive breastfeeding after adjusting for measures of socioeconomic status and adverse housing conditions at birth. RESULTS: Duration of exclusive breastfeeding in infancy was significantly associated with H. pylori seropositivity (odds ratio per 30 days, 0.88; 95% confidence interval, 0.78 to 0.98). The significant protective trend was only seen among men (odds ratio per 30 days, 0.78; 95% confidence interval, 0.65 to 0.95), with no significant effect seen among women. CONCLUSION: Increased duration of exclusive breastfeeding in infancy may have a long-term protective effect against chronic H. pylori infection and hence the risk of gastric carcinoma. Although further research is required, particularly as to why a significant effect was only seen among men, the results provide additional support for the concept that breastfeeding may have long-term influences on health and that human milk is the ideal complete first diet for human infants. PMID- 16254520 TI - Prevalence of celiac disease in Brazilian children and adolescents with type 1 diabetes mellitus. AB - OBJECTIVE: A previously unrecognized high prevalence of celiac disease (CD) has been found by screening among European and North American patients with type 1 diabetes mellitus (DM 1). The prevalence of CD among Brazilian children with DM1 is not known. This study was conducted to determine the prevalence of CD in Brazilian children and adolescents with DM 1. METHODS: One hundred and four children and adolescents with DM 1 (52 males and 52 females; age range 22 months 19 years) and 105 age and gender-matched control participants were screened for CD using the IgA anti-endomysial antibody test (IgA-EmA) and total serum IgA. A small bowel biopsy was performed in all patients with positive IgA-EmA. RESULTS: Nine of 104 diabetic patients (8.7%) had a positive IgA-EmA. Biopsies were normal in four patients, two had partial or subtotal villous atrophy with elevated intraepithelial lymphocyte (IEL) counts, and three showed partial villous atrophy but with IEL counts under the maximum limit adopted (40 IEL/100 enterocytes). EmA IgA positive patients had mild, non-specific gastrointestinal complaints including dyspepsia, abdominal pain, flatulence and constipation. All control participants had negative results for IgA-EmA. CONCLUSION: The prevalence of CD in a group of Brazilian pediatric DM 1 patients was at least 4.8%, a prevalence comparable to European and North American studies. The high prevalence of CD among DM 1 emphasizes the need for routine screening in all countries including Brazil. PMID- 16254521 TI - Double-blind randomized evaluation of clinical and biological tolerance of polyethylene glycol 4000 versus lactulose in constipated children. AB - OBJECTIVES: To assess the safety of a polyethylene glycol (PEG) 4000 laxative without additional salts in pediatric patients. STUDY DESIGN: This was a 3-month multicenter, randomized, double-blind, double-dummy, lactulose-controlled, parallel study enrolling 96 ambulatory constipated children aged 6 months to 3 years, treated daily with 4-8 g PEG or 3.33 g-6.66 g lactulose. Total protein, albumin, iron, electrolytes, and vitamins B9 (folates), A and D (25OHD3) were measured in blood before and after treatment (day 84) in a central laboratory. RESULTS: The percentage of children with at least one value out of normal range at day 84 with respect to baseline status (with or without at least one value out of normal range), i.e. the primary endpoint, was 87% and 90% in the PEG and lactulose groups, respectively, without any difference between groups. The whole blood parameters showed no qualitative or quantitative treatment-related changes. Vitamin A values were above normal range in 56% and 41% of children at baseline versus 33% and 36% at day 84 in the PEG and lactulose groups, respectively. Iron values were similarly under normal range in 47% and 51% at baseline versus 42% and 51% at day 84. Clinical tolerance was similar for both treatments except for vomiting and flatulence, which were significantly higher with lactulose. Significantly higher improvements were evidenced with PEG regarding stool consistency, appetite, fecaloma and use of additional laxatives. CONCLUSION: This 3-month study in 96 constipated children aged 6 months to 3 years confirms the long-term tolerance of PEG 4000 in pediatrics and indicates a PEG efficacy similar to or greater than that of lactulose. PMID- 16254522 TI - Liver histology and alanine aminotransferase levels in children and adults with chronic hepatitis C infection. AB - BACKGROUND: Chronic hepatitis C is often a mild disease in children, but whether this is related to younger age or shorter duration of infection is unclear. Histologic severity has been shown to correlate with duration of infection regardless of age. OBJECTIVES: We compared histologic findings in children and adults with chronic hepatitis C while controlling for sex, duration of infection, hepatitis C virus (HCV)-RNA level, and genotype. METHODS: Twenty-one children and 52 adults whose infection was less than 20 years in duration and who had undergone a liver biopsy were included. Two blinded liver pathologists reviewed the liver biopsies and scored inflammatory activity and fibrosis using the modified Knodell scoring system. RESULTS: The groups were the same with respect to HCV-RNA level (P=0.8), and genotype (P=0.6) but differed in duration of disease (P=0.01) and sex composition (P=0.005). Covariate analysis showed no influence of genotype, duration of infection, or HCV-RNA level on outcome. In controlling for sex, children had significantly milder liver disease and alanine aminotransferase (ALT) elevations. CONCLUSIONS: With equal duration of infection, HCV-RNA level, and genotype, children have lower serum ALT levels and less severe liver disease than adults infected with HCV. PMID- 16254523 TI - Safety of percutaneous liver biopsy in infants less than three months old. AB - OBJECTIVES: To evaluate the safety, outcomes, and complications of percutaneous liver biopsies (PLB) in infants aged 0 to 3 months. METHODS: We retrospectively reviewed the hospital records of all infants less than 3 months old who underwent PLB at Children's Memorial Hospital between July 1, 1997 and June 30, 2004 for complications surrounding the procedure and risk factors that might lead to complications. RESULTS: Sixty-six PLBs were performed in 63 infants. Most patients tolerated the procedure without complications. Twelve complications were recorded, for an overall complication rate of 18%. Of these, five were directly related to the procedure, and seven were sedation related. Three patients experienced a drop in hemoglobin greater than 2 gm/dL, one patient developed a bile leak, and one developed a skin hematoma. Seven patients had respiratory difficulty related to sedation, which manifested as increased work of breathing or decreased respiratory rate with depression in pulse oximetry. CONCLUSION: We conclude that PLB in young infants is associated with a somewhat higher risk of complications than in older children, particularly complications related to sedation. PMID- 16254524 TI - Pancreatic development in newborn guinea pigs fed intact or low-hydrolyzed protein formulas. AB - AIM: To evaluate pancreatic development in newborn guinea pigs fed since birth intact or low-hydrolyzed protein formulas compared with breast milk. METHODS: Forty-five newborn guinea pigs were allocated to three feeding regimens: breast milk (n=15) and two isocaloric isonitrogen milk formulas containing intact (n=15) or low-hydrolyzed proteins (n=15). Body weight and food consumption were recorded every day. After 8 days, one third of pups from each group was killed, and the remaining animals were weaned. Another third was killed on day 14, and the remainders were killed on day 20. Zymogen storage was evaluated on pancreatic sections, whereas DNA and RNA concentrations were measured by a fluorometric method. RESULTS: Compared with breast fed pups, both groups of artificially fed animals showed lower weight gain during the first 2 weeks of life but not after weaning. Both formulas fed groups had significantly lower amount of zymogen granules in pancreatic acinar cells at 8 and 14 days of life. This reduction was still present at day 20 in intact protein formula but not in low-hydrolyzed protein formula fed animals in which higher RNA/DNA ratio was also observed compared with breast fed pups. CONCLUSION: In newborn guinea pigs, artificial feeding is associated with reduced zymogen storage at days 8 and 14 of life. After weaning, cellular content of zymogen granules is comparable with breast fed pups only in low-hydrolyzed protein formula fed animals, even in the presence of some evidence of pancreatic hypoplasia. PMID- 16254525 TI - The effect of ribavirin on bone density in patients with chronic hepatitis C treated with interferon-ribavirin therapy. AB - OBJECTIVE: The objective of this study was to investigate the effects of ribavirin on bone mineral metabolism in patients with chronic hepatitis C who had been treated with interferon and ribavirin. METHODS: Twenty patients (3 female, 17 male) with chronic hepatitis C were enrolled. Age range was 6 to 15 years (mean+/-SD, 11.15+/- 2.3 years). Thirteen patients received combined interferon alpha-2b and ribavirin therapy (Group 1), and 7 patients received only interferon alpha-2b (Group 2). Both groups were treated for 12 months. Bone mineral density, z-scores and biochemical bone markers were evaluated in both groups before and after treatment. RESULTS: There were no significant differences between the groups in age or gender. Mean lumbar vertebral bone mineral density and mean z scores in groups 1 and 2 before and after treatment were not significantly different. In both groups, serum and urinary biochemical values and bone markers were all normal and there were no differences between the pretreatment and post treatment values. CONCLUSION: Contrary to studies in adults, we did not find any ribavirin-dependent changes related to bone mineral metabolism in our pediatric study groups. Further studies are needed to obtain more detailed information about the effects of ribavirin on bone mineral density. PMID- 16254526 TI - Serum ghrelin concentration and weight gain in healthy term infants in the first year of life. AB - OBJECTIVES: Ghrelin, a recently discovered hormone mainly secreted by the stomach, has several metabolic functions including regulation of food intake, energy homeostasis and body weight. There are few studies on this hormone in healthy infants during the first year of life. The aim of this study was to examine the correlations between ghrelin and weight gain in healthy term infants in the first year of life. METHODS: 104 healthy term infants aged 0 to 12 months were included in a cross-sectional study. Anthropometric measurements were assessed and mean weight gain was calculated. Serum ghrelin concentrations have been determined at least 3 hours after feeding by radioimmunoassay test. RESULTS: Ghrelin concentrations were correlated negatively to weight gain (r=-0.302; P=0.003) and positively to age (r = 0.412; P < 0.001), weight (r = 0.374; P < 0.001) and length (r=0.387; P<0.001). In breastfed infants a statistically significant negative correlation between ghrelin concentration and infant weight gain (r=-0.407; P=0.001) was observed, whereas in formula-fed infants this correlation was not statistically significant (r=-0.067; P=0.719). CONCLUSIONS: The negative correlation observed between ghrelin concentration and infant weight gain suggests that ghrelin might also play a role in the regulation of body weight in healthy infants with a physiologic energy balance. Further studies are needed to clarify how ghrelin might be involved in both short-term and long-term energy balance. PMID- 16254527 TI - Widespread use of soy-based formula without clinical indications. AB - OBJECTIVES: In view of reports of the growing popularity of soy-based formula for infants, we examined soy consumption and its possible overuse during early infancy in central Israel. METHODS: Mothers of 1,803 infants aged 2, 4, 6 and 12 months attending well-baby clinics participated in a telephone survey covering background data, rate, duration, and pattern of soy-based formula use and the reasons for its initiation. The reasons were grouped into those based on the recommendations of the medical personnel and those based on mothers' initiative, and evaluated according to infants' age at soy-based formula initiation (0 to 1, 2 to 4 and 5 to 12 months). The symptoms that prompted soy-based formula use were assessed quantitatively. RESULTS: The rate of soy-based formula use was 10.4% at 2 months and 31.5% at 12 months (P<0.001); 70.6%+/- 2.7% of the infants were given soy for>6 months. Regardless of infants' age, the role of the mothers in the decision to use soy-based formula was greater than that of the medical personnel, and increased significantly with age (chi for trend=0.018). A suspicion of cow's milk allergy was responsible for only 10.9% (7/64) of all soy initiations in infants aged 5 to 12 months. In all ages, occasional symptoms, mainly diarrhea (33.3%) and colic (19.8%), were the leading cause for recommending soy-based formula by medical personnel, whereas the personal preference without clinical justification was the leading cause among mothers. CONCLUSIONS: The use of soy-based formula in central Israel is extensive and continues for long periods, with rates far beyond clinical indications. Mothers play a greater role than medical personnel in the decision to initiate soy-based formula. PMID- 16254528 TI - Highly destructive perianal crohn disease. PMID- 16254529 TI - Magnetic foreign body ingestions leading to duodenocolonic fistula. PMID- 16254531 TI - Hepatocellular carcinoma in children with Alagille syndrome. PMID- 16254530 TI - Colorectal spirochetosis in a child with rectal bleeding: case report and literature review. PMID- 16254532 TI - Intramural esophageal hematoma (IEH). PMID- 16254534 TI - Re: neonatal and infant feeding: effect on bone density at 4 years. PMID- 16254535 TI - Lymphoma after Imuran and 6-MP: a new look. PMID- 16254536 TI - A pilot test of the addition of zinc to the current case management package of diarrhea in a primary health care setting. AB - Zinc is recommended for the treatment of acute diarrhea in children but the effect of its introduction on drug and oral rehydration solution use is unclear. Government care providers, private practitioners and community workers were trained to distribute zinc and oral rehydration solution to children seeking care for diarrhea. Periodic surveys showed that village-based workers became a common source of diarrhea treatment and private practitioners were used less. Zinc was used in approximately half of the episodes; the prescription and use rates of oral rehydration solution packets increased from 7% at baseline to 44.9% 6 months later. Reduction in use of drugs during diarrhea ranged from 34% for tablets to 64% for injections 6 months later. The cost of treatment to families declined significantly. These findings need confirmation in a randomized controlled trial. PMID- 16254537 TI - Knowledge about sexually transmitted diseases in women among primary care physicians. AB - OBJECTIVES: Little is known about sexually transmitted disease (STD) knowledge of primary care providers. The objectives of this study were to determine the knowledge about the management of STDs among primary care physicians and to identify physician characteristics associated with possession of STD knowledge. STUDY: A self-administered questionnaire was mailed to a random sample of 1600 obstetrician/gynecologists, pediatricians, family physicians, and internists practicing in Pennsylvania. Information on physician and patient demographics was gathered, and we assessed knowledge and practice patterns concerning the management of STDs in young women. RESULTS: Physician knowledge regarding the evaluation and management of women with or at risk for STDs was associated with female gender (odds ratio [OR]: 2.1; 95% confidence interval [CI]: 1.4-3.2), age < or =40 (OR: 2.3; 95% CI: 1.4-3.6), and metropolitan practice location (OR: 1.7; 95% CI: 1.1-2.6). Familiarity with the Center for Disease Control and Prevention's (CDC's) STD treatment guidelines was independently associated with STD knowledge (OR: 2.0; 95% CI: 1.2, 3.3). Physicians with good STD knowledge were more likely to report routinely screening at-risk women for Chlamydia trachomatis (OR: 3.9; 95% CI: 2.3-6.8). CONCLUSIONS: Inadequacies in physician knowledge may serve as a barrier to the appropriate diagnosis and treatment of STDs. Interventions to improve STD management practices should include continuing medical education and distribution of CDC's STD treatment guidelines to primary care providers. PMID- 16254538 TI - Risk factors for bacterial vaginosis in women at high risk for sexually transmitted diseases. AB - OBJECTIVE AND GOAL: Bacterial vaginosis (BV) is extremely common and is associated with adverse obstetrical and gynecological outcomes. The etiology of the microbiologic changes is unknown. The objective of this study was to determine risk factors associated with incident BV. STUDY: 96 women without BV were followed prospectively for 1 year for the development of BV. Thirty-seven of their male partners were also studied. RESULTS: The incidence rate of BV was 2.33/person-year (95% CI, 1.63-2.50). Median time to development of BV was 81 days. Incident BV was significantly associated with exposure to a new sexual partner (RR, 1.13; 95% CI, 1.02-1.25; P = 0.02) and frequency of vaginal sex since last visit (RR, 1.07; 95% CI, 1.01-1.15; P = 0.03). Use of condoms with occasional partners was protective (RR, 0.80; 95% CI, 0.67-0.98; P = 0.003). In multivariate analysis, sex with a new partner since the prior visit was the only behavior found to be significantly associated with incident BV (RR, 1.74; 95% CI, 1.05-2.87; P = 0.03). CONCLUSION: These data support the sexual transmission of BV. PMID- 16254539 TI - Reassessing a large-scale syphilis epidemic using an estimated infection date. AB - OBJECTIVES: Timely ascertainment of syphilis cases is critical to initiating disease-control measures. Epidemic curves typically use the report date and may introduce lag-time bias into assessment. GOAL: To reassess a large syphilis epidemic using an imputed infection date. STUDY: We compared 2 types of epidemic curves-1 based on report date and 1 on estimated infection date-using the large 1993-2003 Baltimore epidemic as our model. RESULTS: In general, the shape of the report curves did not accurately reflect the shape of the corresponding infection curves during the growth period (period of largest increase in incidence); during the hyperendemic period (period of highest incidence), peaks in report curves did not follow peaks in the infection curve by the appropriate lag time. There was a tendency for reporting data to underestimate infections during the growth period and overestimate infections during the hyperendemic period. A sensitivity analysis showed similar trends regardless of the length of stage-specific incubation period used. CONCLUSIONS: Lag-time bias may be present when using epidemic curves based on report dates. Health departments should consider using an estimated infection date. PMID- 16254540 TI - Do participant characteristics influence the effectiveness of behavioral interventions? Promoting condom use to women. AB - OBJECTIVES: This study assessed whether participant baseline characteristics modified the effects of a skill-based intervention promoting condom use. STUDY: The randomized, controlled trial enrolled 427 women from a sexually transmitted disease clinic in Birmingham, Alabama. The main outcome measures: consistent (100%) and problem-free (correct, no breakage or slippage) condom use were verified by sexual diary self-report and contraceptive product counts. RESULTS: The enhanced intervention group had a 60% higher consistent condom use rate compared to the basic group (risk ratio [RR], 1.6; 95% confidence interval [CI], 1.4-1.8). There was no statistically significant difference between groups in relationship to problem-free, consistent use (RR, 1.0; 95% CI, 0.9-1.1). A binomial regression analysis identified the following factors as significant modifiers of intervention effectiveness on consistent condom use: intention to use condoms next time, early-age sexual debut, marital status combined with place of intercourse, and substance use before sex. CONCLUSIONS: The results suggest that participant baseline characteristics can be modifiers of intervention effectiveness. PMID- 16254541 TI - A randomized trial of clinician-delivered interventions promoting barrier contraception for sexually transmitted disease prevention. AB - OBJECTIVE: The objective of this study was to compare 2 interventions promoting condoms and vaginal microbicides to prevent sexually transmitted disease (STD). STUDY: Women (N = 427) attending an STD clinic were randomly assigned to 2 clinician-delivered interventions and followed up monthly to assess condom/microbicide use and incidence of gonorrhea, chlamydia, and syphilis. RESULTS: During follow up, condom use rates were 69% (enhanced) and 49% (basic) and microbicide use rates were 44% and 29%, respectively. STD rates did not significantly differ between intervention groups. Perfect condom use (regardless of intervention arm) was associated with a 3-fold decrease in STD rates (relative risk [RR], 0.3; 95% confidence interval [CI], 0.1-0.8). Using a vaginal microbicide during > or =50% of the acts of intercourse was associated with reduced STD rates (RR, 0.5; 95% CI, 0.3-1.0) across intervention groups and condom use categories. CONCLUSIONS: The enhanced intervention increased use of condoms and vaginal microbicide; however, STD rates did not decrease because a protective effect was seen only among perfect barrier users, and the enhanced intervention only modestly increased perfect use. PMID- 16254542 TI - Evaluation of sexually transmitted diseases/human immunodeficiency virus intervention programs for sex workers in Calcutta, India. AB - BACKGROUND AND OBJECTIVE: The Sonagachi Project in Calcutta, India, organized sex workers to improve working conditions. GOAL: To compare rates of sexually transmitted diseases between the Sonagachi Project and other areas in which only the National AIDS Control Organization (NACO) interventions were implemented. STUDY: A cross-sectional survey of randomly selected female sex workers. RESULTS: There was no difference in the prevalence of all STDs between the 2 areas; both were lower than reported in other surveys in 1992. Analysis using propensity scores also failed to demonstrate any difference. The number of preventive activities was similar in the Sonagachi and NACO-only areas but was more prevalent than in 1992. Sex workers in the Sonagachi area had better treatment seeking behavior and attitudes. Both the Sonagachi and NACO strategies have resulted in lower STD rates, but the Sonagachi Project also increased the proportion who had an optimistic attitude and increased prevention and treatment seeking behavior. PMID- 16254543 TI - Alcohol use by men is a risk factor for the acquisition of sexually transmitted infections and human immunodeficiency virus from female sex workers in Mumbai, India. AB - OBJECTIVE: We investigated whether men who were under the influence of alcohol when visiting female sex workers (FSW) were at greater risk for sexually transmitted infections (STI) and human immunodeficiency virus (HIV). STUDY: A cross-sectional analysis using baseline data from a randomized controlled trial of an HIV prevention intervention for high-risk men in Mumbai, India. RESULTS: The overall HIV prevalence among 1741 men sampled was 14%; 64% had either a confirmed STI or HIV; 92% reported sex with an FSW, of whom 66% reported having sex while under the influence of alcohol (SUI). SUI was associated with unprotected sex (odds ratio [OR]: 3.1; 95% confidence interval [CI], 2.3-4.1), anal sex (OR: 1.5; 1.1-2.0), and more than10 FSW partners (OR: 2.2; 1.8-2.7). SUI was independently associated with having either an STI or HIV (OR: 1.5; 1.2-1.9). CONCLUSION: Men who drink alcohol when visiting FSWs engage in riskier behavior and are more likely to have HIV and STIs. Prevention programs in India need to raise awareness of this relationship. PMID- 16254544 TI - Sex with women as a risk factor for herpes simplex virus type 2 among young men who have sex with men in Baltimore. AB - BACKGROUND: Herpes simplex virus type 2 is common among MSM and is a risk factor for transmission of HIV. The findings of studies investigating the relationship between infection with HSV-2 and number of sex partners among MSM are inconsistent and rarely distinguish between male and female partners. GOAL: To determine the prevalence and risk factors for infection with HSV-2, including the number and gender of sex partners, in a group of MSM in Baltimore, MD. STUDY: This was a cross-sectional study among young MSM in Baltimore. RESULTS: Of the blood samples from 824 participants, 19.3% had HSV-2 antibodies. After adjusting for known HSV-2 correlates, independent predictors of HSV-2 seropositivity included HIV seropositivity, black race, older age, number of lifetime female sex partners, recent unprotected receptive anal intercourse with a man. CONCLUSIONS: This study suggests that female sex partners may be an important source of HSV-2 infection among young bisexual MSM. After adjusting for known HSV-2 correlates, the number of lifetime female but not male sex partners was independently associated with HSV-2. These results highlight the need for HSV-2 prevention and treatment efforts targeting MSM who also have sex with women. Future investigations of HSV-2 and sexual behavior among MSM need to distinguish between male and female sex partners. PMID- 16254545 TI - Vaginal douching, condom use, and sexually transmitted infections among Chinese female sex workers. AB - BACKGROUND AND OBJECTIVE: Vaginal douching has been hypothesized to increase a woman's risk for human immunodeficiency virus (HIV) infection. However, data on the prevalence of this practice and its association with condom use and sexually transmitted infections (STIs) are limited. STUDY: A cross-sectional survey among 454 female sex workers (FSWs) in a Chinese county. RESULTS: Vaginal douching was reported by 64.7% of the women. The prevalence of self-reported history of STI and that of current STI was 19.4% and 41.5%, respectively. Fifteen percent of the women reported consistent use of condoms with their clients and 8.4% with their regular partners. Vaginal douching was significantly associated with decreased use of condoms (with clients: OR = 0.31; with regular partner(s): OR = 0.22) and increased rate of self-reported STI history (OR = 1.95). However, there was no direct relation between douching and current STI. Over one third of the women believed that douching can prevent STI/HIV. CONCLUSION: Vaginal douching exposes FSWs to a high risk of STI/HIV. Medical professional and public health workers should correct women's misconception about the effectiveness of douching and discourage women from douching through educational activities. PMID- 16254546 TI - The impact of highly active antiretroviral therapy and immunodeficiency on human papillomavirus infection of the oral cavity of human immunodeficiency virus seropositive adults. AB - BACKGROUND: Prevalence of human papillomavirus (HPV)-associated oral condylomas has reportedly increased in HIV-infected individuals since the introduction of highly active antiretroviral therapy (HAART). The relationships between HIV therapy regimen, overall health, and subclinical oral HPV have not been examined. GOAL: To determine oral HPV genotype prevalence and the impact of HAART and health in the HIV+ population. STUDY: An L1 consensus-primer polymerase chain reaction and linear array assay were used to examine the prevalence of 27 HPV genotypes in saliva of 98 HIV+ individuals. Risk assessment variables were compared to oral HPV status. RESULTS: Oral HPV was detected in 37% of HIV+ African American individuals. Caucasians were at greater risk of oral HPV infection than African Americans. Markers of advanced HIV disease did not predict HPV status. Therapy status was associated with HPV detection. CONCLUSIONS: Treatment of HIV, rather than HIV immunosuppression, appears to play a role in oral HPV infections in HIV+ individuals. PMID- 16254547 TI - Temporal trends in sexual behaviors and sexually transmitted disease history among 18- to 39-year-old Seattle, Washington, residents: results of random digit dial surveys. AB - OBJECTIVES: The objectives of this study were to describe sexual behaviors of Seattle residents in 2003-2004 and report changes since 1995. METHODS: We conducted a random digit-dial (RDD) survey among 18- to 39-year-old men and women in 2003-2004. We compared the results with the results of a 1995 RDD survey conducted in the same population. Questionnaire batteries and sampling methods were similar in the 2 surveys. RESULTS: Between 1995 and 2004, the median number of lifetime sex partners increased from 7 to 8. Vaginal douching declined from 70.6% to 25.9% (P = 0.004) among black women; from 8.3% to 5.9% (P <0.05) among Asian American women; and from 15.5% to 2.4% (P <0.05) overall. After adjustment, proportions of women who reported practicing vaginal douching (odds ratio [OR], 0.13; 95% confidence interval [CI], 0.7-0.22), the proportion of respondents who reported a history of any sexually transmitted disease (OR, 0.70; 95% CI, 0.53 0.93) and a history of gonorrhea (OR, 0.48; 95% CI, 0.25-0.89) declined between 1995 and 2004. Conversely, the proportion of respondents who reported only same sex partners (OR, 3.27; 95% CI, 1.33-9.88), condom use at first sex with their most recent sex partner (OR, 1.38; 95% CI, 1.06-1.78), and reported practice of anal sex (OR, 2.01; 95% CI, 1.21-3.48) increased between 1995 and 2004. Among blacks, proportions reporting an age difference of only 2 years with their partners declined from 64.3% to 25.9% (P <0.05), indicating increased age mixing. CONCLUSIONS: Some risk behaviors declined whereas others increased between 1995 and 2004; several trends were divergent between the general population and minority populations. Our data hint at an increasing divergence in risk behaviors and morbidity between minority populations and the general population. Also, anal sex and number of sex partners have increased. These patterns have serious implications for the design, targeting, and implementation of prevention programs. PMID- 16254548 TI - Outbreak of syphilis among HIV-infected patients: descriptive data from a Parisian hospital. PMID- 16254549 TI - Novel mutations in GJA3 associated with autosomal dominant congenital cataract in the Indian population. AB - PURPOSE: Connexin 46 (Cx46) is crucial in the maintenance of lens homeostasis and it is known to be expressed mainly in the terminally differentiated lens fiber cells. The present study aimed to identify the spectrum of mutations in Connexin 46 in the Indian population. METHODS: PCR based Single Stranded Conformational Polymorphism (SSCP) analysis was used to screen sixty probands with nonsyndromic congenital cataract for mutations in the Cx46 gene (GJA3), followed by direct sequencing of samples that showed an electrophoretic shift. Mutation predicted to affect the coding sequence were subsequently analyzed in the entire pedigree. RESULTS: Two novel missense mutations were identified in Cx46. The mutation in Family 1 was characterized as R76G with a total cataract phenotype. A V28M missense mutation was identified in family 2, the cataract phenotype varied in its severity and the age of onset. The mutation was also identified in 2 unaffected individuals of the family and the intrafamilial variation of the disease suggests the possibility of a modifier gene(s) or the effects of environmental factors being involved. The mutation was identified in all the affected members in the family and found to be absent in 400 ethnically matched control chromosomes analyzed. CONCLUSIONS: We conclude that connexin 46 mutations might account for as much as 3.3% of the hereditary congenital cataract in the Indian population. PMID- 16254550 TI - Expression of superoxide dismutase in whole lens prevents cataract formation. AB - PURPOSE: Oxidative damage is a major factor causing cataracts, which account for almost half of human blindness cases worldwide. In this study, we wished to determine if overexpression of superoxide dismutase (SOD) in intact lenses could prevent cataract formation induced by oxidative stress. METHODS: Fresh, intact lenses from 6-week-old male/female Sprague Dawley rats were incubated with plasmid DNA encoding the human SOD1 (Cu/Zn-SOD) gene at 37 degrees C in a CO2 cell culture chamber with 95% air and 5% CO2. SOD1 expression was determined by western blotting and SOD enzyme activity. Lenses with or without overexpression of SOD1 were treated with H2O2 and cataract formation was examined. SOD1 regulation of protein kinase Cgamma (PKCgamma) was determined by PKCgamma enzyme activity assay. Intact lens gap junctions were determined by dye transfer assay. RESULTS: In the lens overexpression system, SOD1 cDNA was fused to EYFP to generate EYFP:SOD1 fusion proteins which allow detection from endogenous SOD1. Incubation of intact lenses with plasmid DNA produced EYFP:SOD1 fusion proteins as determined by western blot using anti-GFP or anti-SOD1 antibodies. This caused significant increases in SOD enzyme activity. Data indicated that SOD1 plasmid DNA can be expressed as a functional enzyme in intact lenses in culture. Lenses overexpressing SOD1 remained clear after H2O2 treatment at 100 muM for 24 h, similar to control. Overexpression of SOD1 diminished the effect of H2O2 on PKCgamma activation and subsequent inhibition of gap junctions, indicating that overexpression of SOD1 may reduce reactive oxygen species (ROS) production, and this would prevent the normal H2O2 effect on cataract formation. CONCLUSIONS: Overexpression of SOD1 in whole lens prevents H2O2-induced oxidative damage (cataract formation) to the lens and subsequent control of gap junctions by protein kinase Cgamma. PMID- 16254551 TI - Effect of excitation wavelength on the Raman spectroscopy of the porcine photoreceptor layer from the area centralis. AB - PURPOSE: Raman microscopy, based upon the inelastic scattering (Raman) of light by molecular species, has been applied as a specific structural probe in a wide range of biomedical samples. The purpose of the present investigation was to assess the potential of the technique for spectral characterization of the porcine outer retina derived from the area centralis, which contains the highest proportion of cone:rod cell ratio in the pig retina. METHODS: Retinal cross sections, immersion-fixed in 4% (w/v) PFA and cryoprotected, were placed on salinized slides and air-dried prior to direct Raman microscopic analysis at three excitation wavelengths, 785 nm, 633 nm, and 514 nm. RESULTS: Raman spectra of each of the photoreceptor inner and outer segments (PIS, POS) and of the outer nuclear layer (ONL) of the retina acquired at 785 nm were dominated by vibrational features characteristic of proteins and lipids. There was a clear difference between the inner and outer domains in the spectroscopic regions, amide I and III, known to be sensitive to protein conformation. The spectra recorded with 633 nm excitation mirrored those observed at 785 nm excitation for the amide I region, but with an additional pattern of bands in the spectra of the PIS region, attributed to cytochrome c. The same features were even more enhanced in spectra recorded with 514 nm excitation. A significant nucleotide contribution was observed in the spectra recorded for the ONL at all three excitation wavelengths. A Raman map was constructed of the major spectral components found in the retinal outer segments, as predicted by principal component analysis of the data acquired using 633 nm excitation. Comparison of the Raman map with its histological counterpart revealed a strong correlation between the two images. CONCLUSIONS: It has been demonstrated that Raman spectroscopy offers a unique insight into the biochemical composition of the light-sensing cells of the retina following the application of standard histological protocols. The present study points to the considerable promise of Raman microscopy as a component-specific probe of retinal tissue. PMID- 16254553 TI - Framework for a fully powered risk engine. PMID- 16254554 TI - EMPReSS: standardized phenotype screens for functional annotation of the mouse genome. PMID- 16254552 TI - The interphotoreceptor retinoid-binding protein (IRBP) of the chicken (Gallus gallus domesticus). AB - PURPOSE: Despite decades of investigation, the function of interphotoreceptor retinoid binding protein (IRBP), the most abundant protein in the interphotoreceptor matrix of vertebrates, remains enigmatic. Roles for IRBP in the visual cycle of rod photoreceptors and in the independent visual cycle of cone photoreceptors have been suggested, yet very little is known of the biology of IRBP in cone-dominant retinas, such as those of diurnal birds. Our aim was to identify and characterize expression of the IRBP of the cone-dominant chicken (Gallus gallus domesticus). METHODS: Chicken IRBP mRNA was identified by PCR cloning. Primary protein structure, genomic organization, and phylogenies were determined through comparative sequence analyses. Expression of IRBP mRNA was characterized by northern analysis and by in situ hybridization on cryosectioned chicken retina. Expression of the IRBP protein was characterized by western blotting and by indirect immunofluorescence on cryosectioned retina and on retinal whole mounts. RESULTS: The chicken IRBP gene encodes a secreted protein with a predicted 1,252 amino acid length. The gene structure for chicken IRBP resembles that of most other vertebrates, with four homologous, modular repeats and introns within only the fourth module. Each module is more homologous with the corresponding module in other species than it is with the remaining chicken modules. Chicken retinal tissue contains a single IRBP mRNA transcript of approximately 4.8 kb and western analysis of chicken retina shows a single major band of 140 kDa. Chicken IRBP mRNA is expressed exclusively by retinal photoreceptor cells and the intensity of the hybridization signal shows light/dark rhythmicity. The IRBP protein is localized to the interphotoreceptor matrix of the chicken retina and to intracellular regions of photoreceptors, with a spatial distribution indicating an association with cone outer segments. CONCLUSIONS: The high degree of conservation of IRBP's primary structure, genomic organization, and cell-specific expression within the retinas of all vertebrates examined to date, including those with cone-dominant retinas, implies a conserved role for IRBP in photoreceptor function and/or health. Expression of chicken IRBP and its mRNA are functionally regulated. This report provides a necessary first step to explore a specific function for IRBP in the cone visual cycle. PMID- 16254556 TI - A ferrireductase fills the gap in the transferrin cycle. PMID- 16254557 TI - Malaria-protective traits at odds in Africa? PMID- 16254558 TI - Insights into the hierarchy of selenium incorporation. PMID- 16254559 TI - RNAi the natural way. PMID- 16254560 TI - Touching base. PMID- 16254562 TI - Donning our fancy coats. PMID- 16254563 TI - Genetic variation in laboratory mice. AB - Characterizing the patterns of genetic variation in an organism provides fundamental insight into the evolutionary history of the organism and defines the scope and nature of studies that must be designed to correlate genotype to phenotype. Given the pre-eminent role of the inbred mouse in biomedical research, considerable effort has been undertaken in recent years to describe more fully the nature and amount of genetic variation among the numerous strains of mice that are in widest use. Here, we discuss recent studies that have contributed to an emerging understanding of the unique variation patterns found in inbred strains of mice and how they have arisen through a combination of natural evolution and human-directed breeding. These preliminary results have ramifications for genetic research into complex biomedical traits and are the basis for the development of future variation resources. PMID- 16254564 TI - The origins and uses of mouse outbred stocks. AB - Outbred mouse stocks, often used in genetics, toxicology and pharmacology research, have been generated in rather haphazard ways. Understanding the characteristics of these stocks and their advantages and disadvantages is important for experimental design. In many studies these mice are used inappropriately, wasting animals' lives and resources on suboptimal experiments. Recently, however, researchers from the field of complex trait analysis have capitalized on the genetics of outbred stocks to refine the identification of quantitative trait loci. Here we assess the most widely used outbred stocks of mice and present guidelines for their use. PMID- 16254565 TI - Current issues in mouse genome engineering. AB - The mouse is the foremost vertebrate experimental model because its genome can be precisely and variously engineered. Now that the mouse genome has been sequenced and annotated, the task of mutating each gene is feasible, and an international cooperation is providing mutated embryonic stem cells and mice as readily available resources. Because these resources will change biomedical research, decisions about their nature will have far-reaching effects. It is therefore timely to consider topical issues for mouse genome engineering, such as the background genotype; homologous, site-specific and transpositional recombination; conditional mutagenesis; RNA-mediated interference; and functional genomics with embryonic stem cells. PMID- 16254566 TI - Strategies for dissecting epigenetic mechanisms in the mouse. AB - Epigenetics generally refers to heritable changes in gene expression that are independent of nucleotide sequence. With complete genome sequences in hand, understanding the epigenetic control of genomes is the next step towards comprehending how the same DNA sequence gives rise to different cells, lineages and organs. Epigenetics also contributes to individual variation in normal biology and in disease states. The mouse provides a unique opportunity to understand how epigenetic differences contribute to both development and disease in a tractable mammalian system. Here we discuss current approaches and protocols used to study epigenetics in the mouse, including loss-of-function studies, mutagenesis screens, somatic cell nuclear transfer, genomics and proteomics. PMID- 16254569 TI - The sexual and reproductive health in men with generalized epilepsy: a multidisciplinary evaluation. AB - This study was specifically aimed to evaluate the sexual and reproductive health in a group of men with generalized epilepsy. In total, 44 men with generalized epilepsy were included in this study, their ages between 18 and 48 years (29.2+/ 9.9) and duration of illness between 2 and 35 years (11.2+/-7.4); 34 patients were treated with conventional antiepileptic drugs (AEDs). Sexological and psychological interviews together with serum total testosterone, E(2), FSH, LH and prolactin were determined. Hyposexuality was diagnosed in 61.4%. Erectile dysfunction (ED) and premature ejaculation represented 70.4 and 66.7%, respectively. Variables such as hyposexuality, seizure duration and its poor control on AEDs were significantly associated with depressive symptoms. Compared to the normal control group, all patients reported elevated E(2) levels (P<0.001), 10 had FSH (n=4) and LH (n=6) levels exceeding that of the normal range for controls and two had hyperprolactinemia. Although the patients' mean value of total testosterone remained within the normal range, but it was significantly lower in hyposexual men compared to nonhyposexual (P<0.002), only two epileptic patients had markedly reduced level of total testosterone beyond normal control levels. This study strongly supports that: (1) The risk of hyposexuality and reproductive disturbances is high in epileptic patients with GTC convulsions despite the AEDs utilized. The risk for SD is further increased by poor seizure control and the frequently accompanied depressive manifestations. (2) It is possible that elevated E2 could increase the risk of SD by reducing active testosterone through negative feedback and the reduction of active testosterone could increase seizure intractability to antiepileptic medications. PMID- 16254570 TI - Downregulation of progesterone biosynthesis in rat granulosa cells by adlay (Coix lachryma-jobi L. var. ma-yuen Stapf.) bran extracts. AB - Adlay (Coix lachryma-jobi L. var. ma-yuen Stapf.) has long been used as a traditional Chinese medicine for dysfunctions of the endocrine system and inflammation conditions. However, the effect of adlay seed on the endocrine system has not yet been reported. In the present study, the effects and the mechanisms of methanolic extract of adlay bran (ABM) on progesterone synthesis in rat granulosa cell were studied. ABM was further partitioned with different solvents including water, 1-butanol, ethyl acetate and n-hexane. Four subfractions named ABM-Wa (water fraction), ABM-Bu (1-butanol fraction), ABM-EA (ethyl acetate fraction) and ABM-Hex (n-hexane fraction) were obtained. ABM-Bu was further fractionated using Diaion HP-20 resin column chromatography with gradient elution. Granulosa cells were prepared from pregnant mare serum gonadotropin-primed immature female rats and challenged with different reagents including human chorionic gonadotropin (hCG 0.5 IU/ml), forskolin (10 microM), 8 bromo-adenosine-3',5'-cyclic monophosphate (8-Br-cAMP, 1 mM), A23187 (10 microM), phorbol 12-myristate 13-acetate (PMA, 0.01 microM), 25-OH-cholesterol (0.1-10 microM) and pregnenolone (0.1-10 microM) in the presence or absence of ABM-Bu (100 microg/ml). The functions of steroidogenic enzyme including protein expression of the steroidogenic acute regulatory protein (StAR) and cytochrome P450 side-chain cleavage enzyme (P450scc) protein were investigated. Expressions of both P450scc and StAR mRNA have also been explored. We found that ABM decreased progesterone production via an inhibition on (1) the cAMP-PKA and PKC signal transduction pathway, (2) P450scc and 3beta-hydroxysteroid dehydrogenase (3beta-HSD) enzyme activity, (3) P450scc and StAR protein and mRNA expressions and (4) the phosphorylation of ERK1/2 in rat granulosa cells. PMID- 16254567 TI - Mice in the world of stem cell biology. AB - The ability of embryos to diversify and of some adult tissues to regenerate throughout life is directly attributable to stem cells. These cells have the capacity to self-renew-that is, to divide and to create additional stem cells-and to differentiate along a specific lineage. The differentiation of pluripotent embryonic stem cells along specific cell lineages has been used to understand the molecular mechanisms involved in tissue development. The often endless capacity of embryonic stem cells to generate differentiated cell types positions the field of stem cells at the nexus between developmental biologists, who are fascinated by the properties of these cells, and clinicians, who are excited about the prospects of bringing stem cells from bench to bedside to treat degenerative disorders and injuries for which there are currently no cures. Here we highlight the importance of mice in stem cell biology and in bringing the world one step closer to seeing these cells brought to fruition in modern medicine. PMID- 16254571 TI - Alterations in grip strength during male sexual arousal. AB - Although it is known that alterations in grip strength occur under a number of conditions, little is known about relationships between grip strength and sexual arousal. This relationship was investigated in 30 healthy heterosexual males, who viewed both erotic and nonerotic videos. A questionnaire was used to assess the extent of sexual arousal. The grip strengths of both hands were measured with a five-position (P1-P5) dynamometer, before and after watching the videos. After watching the erotic video, there was a statistically significant reduction in grip strength for the P2 position, with nonsignificant overall reductions in grip strength for all other positions tested. No such effect was observed in control tests. The results indicate that during sexual arousal, the neural system is likely to reduce the output to muscles not directly related to sexual function, presumably to enhance the physiological responses of sexual arousal. PMID- 16254572 TI - Acute and long-term proteome changes induced by oxidative stress in the developing brain. AB - The developing mammalian brain experiences a period of rapid growth during which various otherwise innocuous environmental factors cause widespread apoptotic neuronal death. To gain insight into developmental events influenced by a premature exposure to high oxygen levels and identify proteins engaged in neurodegenerative and reparative processes, we analyzed mouse brain proteome changes at P7, P14 and P35 caused by an exposure to hyperoxia at P6. Changes detected in the brain proteome suggested that hyperoxia leads to oxidative stress and apoptotic neuronal death. These changes were consistent with results of histological and biochemical evaluation of the brains, which revealed widespread apoptotic neuronal death and increased levels of protein carbonyls. Furthermore, we detected changes in proteins involved in synaptic function, cell proliferation and formation of neuronal connections, suggesting interference of oxidative stress with these developmental events. These effects are age-dependent, as they did not occur in mice subjected to hyperoxia in adolescence. PMID- 16254573 TI - BCL-xL: time-dependent dissociation between modulation of apoptosis and invasiveness in human malignant glioma cells. AB - Conditionally BCL-xL-overexpressing LNT-229 Tet-On glioma cell clones were generated to investigate whether the 'antiapoptosis phenotype' and the 'motility phenotype' mediated by BCL-2 family proteins in glioma cells could be separated. BCL-xL induction led to an immediate and concentration-dependent protection of LNT-229 cells from apoptosis. BCL-xL induction for up to 3 days did not result in altered invasiveness. In contrast, long-term BCL-xL induction for 21 days resulted in increased transforming growth factor-beta2 expression, and in metalloproteinase-2 and -14 dependent, but integrin independent, increased invasiveness. Withdrawal of doxycycline (Dox) abolished the protection from apoptosis whereas the 'invasion phenotype' remained stable. Dox stimulation of BCL-xL-inducible LNT-229 cells conferred infiltrative growth to BCL-xL-positive glioma cells in vivo and reduced the survival of tumor-bearing mice. These data allow to dissect a direct antiapoptotic action of BCL-xL from an indirect effect, presumably mediated by altered gene expression, which modifies tumor cell invasiveness in vitro and in vivo. PMID- 16254574 TI - Importance of zinc in the elderly: the ZENITH study. AB - The elderly are at nutritional risk as a result of multiple physiological, social, psychological, and economic factors. Physiological functions naturally decline with age, which may influence absorption and metabolism. Social and economic conditions can adversely affect dietary choices and eating patterns. However, at the same time, the nutrient needs of the elderly for certain nutrient (such as vitamins, minerals, proteins) is higher than for younger adults. This article reviews the importance of zinc (Zn) in elderly people, particularly for behavioural and mental function, micronutrient status, immune and antioxidant system, and bone metabolism. PMID- 16254576 TI - Positive and negative mood in the elderly: the ZENITH study. AB - OBJECTIVE: To assess the quality of positive and negative affect (mood) in an ageing European sample. BACKGROUND: Mood quality has important implications for both physical and mental wellbeing. Poor quality moods are associated with deficits in the diverse areas of cognitive function, health, and social relationships. The ageing process presents a number of potential challenges to successful mood regulation that could have wider implications. DESIGN AND PARTICIPANTS: The current study examines the quality of positive and negative affect in 387 healthy participants from three European countries. Moods were measured four times a day for 4-7 d with the Positive and Negative Affect Schedule (PANAS) mood scales. Measures of zinc (Zn) status were taken also. SETTING: Two centres concentrated on 55-70 yr olds (Coleraine, N.Ireland, n = 93 and Clermont-Ferrand, France, n = 95), and two centres concentrated on 70-87 yr olds (Rome, Italy, n = 108, and Grenoble, France, n = 91). RESULTS: Positive affect scores for the centre in Rome were significantly (P < 0.01) lower than for the other three centres, and the Grenoble centre had significantly (P < 0.05) higher scores on negative affect than the other three centres. Mood was not related to measures of zinc status (all Ps > 0.05). CONCLUSIONS: The two centres with the oldest participants showed deficits in mood quality that may have implications for broader well-being. PMID- 16254575 TI - Health and lifestyle characteristics of older European adults: the ZENITH study. AB - OBJECTIVE: To describe health and lifestyle factors of participants in the ZENITH study. DESIGN: A prospective multicentre intervention study employing a randomised double-blind design. PARTICIPANTS: Community dwelling older adults (n = 387), aged 55-87 y were recruited from regions in France, Italy and the UK. INTERVENTION: A self-report questionnaire comprising socio-demographic variables, dietary habits, physical activity in the home, at work and recreation. RESULTS: Participants differed with regards dietary habits and physical activity for each region. Recreational activity was higher in France and women generally tend to perform less hours of recreational activity per week than men. CONCLUSIONS: The differences found for these regions of Europe in relation to lifestyle factors will affect health and well-being within these countries and may mediate the impact of zinc supplementation on various biological and psychological parameters. PMID- 16254577 TI - Cognitive function in healthy older European adults: the ZENITH study. AB - OBJECTIVE: Baseline data are reported from a study of the effects of zinc supplementation on cognitive function in older adults as assessed by the CANTAB computerised test battery. DESIGN: This is a multicentre prospective intervention study employing a randomised double-blind design. SETTING: European community based study. PARTICIPANTS: There are 387 healthy adults aged 55-87 y from centres in France, Italy and Northern Ireland. INTERVENTIONS: Measures of visual memory, working memory and attention were obtained at baseline (prior to supplementation). RESULTS: Younger adults (<70 y) performed significantly better than older adults (>70 y) on all tests, with minimal differences between centres. In addition, men outperformed women on tests of spatial span, pattern recognition memory and reaction times, although these gender differences varied somewhat between centres. CONCLUSIONS: The results are generally consistent with previous age- and gender-related effects on cognitive functioning. PMID- 16254578 TI - Zinc status and taste acuity in older Europeans: the ZENITH study. AB - BACKGROUND: Age-related decline in taste acuity may be both a cause and an effect of depleted zinc and/or increased zinc requirement. OBJECTIVE: The aim of this study was to explore associations between zinc status and taste acuity in healthy older European adults aged 55-90 y. SAMPLE: Volunteers were recruited within Italy (n = 108 aged 70-90 y), the United Kingdom (UK) (n = 93 aged 55-70) and two regions of France (n = 186), Grenoble (aged 70-90 y) and Clermont-Ferrand (aged 55-70 y). METHODS: A signal detection theory approach was adopted, employing a three-alternative, forced-choice procedure. The data were converted to R-indices and bivariate correlations were computed to explore relationships between serum zinc, erythrocyte zinc and taste acuity. ANOVA was undertaken to determine regional differences in zinc status. RESULTS: Higher erythrocyte zinc status was associated with better acuity for salt (sodium chloride) taste in the sample as a whole (P = 0.012) (n = 385). Higher serum zinc levels were associated with greater sensitivity to sour taste (citric acid) (P = 0.015) only in the older groups (aged 70-90 y). There were no apparent associations between serum or erythrocyte zinc status and acuity for bitter (quinine) or sweet (sucrose) tastes irrespective of age. CONCLUSION: These results agree with those previously suggesting that age-related detriment in sensitivity for salt taste may be associated with depleted zinc. PMID- 16254579 TI - Zinc intake and status in middle-aged and older European subjects: the ZENITH study. AB - OBJECTIVE: Inadequate intakes of micronutrients in elderly negatively affect the nutritional status. Zinc is an essential micronutrient in the elderly, especially in relation to its impact on immune function, bone mass, cognitive function and oxidative stress. However, data are lacking on zinc intake and status during normal ageing. In this study, we evaluate the intake and status of zinc in late middle-aged and older free-living subjects. DESIGN: Dietary zinc intake and zinc status in 188 middle-aged subjects from Clermont-Ferrand (Fr) and Coleraine (UK), and in 199 older subjects from Grenoble (Fr) and Roma (It) were assessed at the entry in the ZENITH study. RESULTS: In relation to the zinc RDA for people older than 55 y, zinc intakes in most of the middle-aged and older subjects (more than 96%) in the present study were adequate. Older people had significantly lower (P < 0.01) energy intakes as compared to middle-aged. Zinc intake expressed per MJ was also significantly (P < 0.01) higher in older people compared to middle-aged. Erythrocyte and urinary zinc concentrations were significantly (P < 0.001) higher in middle-aged subjects compared to older ones. The prevalence of biological Zn deficiency in free-living ageing European people was low (<5%). CONCLUSION: The results of the present study showed a relatively low prevalence of zinc deficiencies in healthy free-living late middle-age and older subjects. These results should be useful for health professionals to have reference data on zinc intake and status for a healthy ageing. PMID- 16254580 TI - Estimation of intake and status of vitamin A, vitamin E and folate in older European adults: the ZENITH. AB - OBJECTIVE: To report selected dietary intake and vitamin status at baseline of volunteers participating in the ZENITH study and the correlation of vitamin status with zinc. DESIGN: A multicentre prospective intervention study employing a randomised double-blind design. SETTING: Clermont-Ferrand, Theix (France), Coleraine (Northern Ireland), Grenoble (France), Rome (Italy). PARTICIPANTS: In total, 387 healthy middle-aged (55-70 y) and older (70-87 y) men and women participated in the study. METHODS: Dietary intake was assessed by means of a validated 4-d recall record. Fasting blood samples were simultaneously analysed for retinol and alpha-tocopherol by the HLPC method. Erythrocyte folates were measured by a competitive immunoassay with direct chemiluminescence detection on an automatised immunoanalyser. RESULTS: In all centres, men had a significantly (P < 0.0001) higher mean nutrient intake than women. Comparison between age groups showed that older individuals had significantly lower intakes of macro- and selected micronutrients than middle-aged subjects (P < 0.0001). A high fat intake (from 36 to 40% of total energy) was observed in all examined groups. In relation to biochemical measures of vitamin status, all parameters were above their respective cut-off values for normality and, thus, none of the subjects had biochemical evidence of deficiency of these selected vitamins. A moderate correlation was found with plasma vitamin A and serum zinc (r = 0.12, P < 0.05) or red blood cell zinc (r = 0.12, P < 0.01) and with erythrocyte folates and red blood cell zinc (r = 0.11, P < 0.05). CONCLUSIONS: There were only moderate differences in the nutrient intake of the ZENITH study volunteers among the four European centres. Their biochemical status for retinol, alpha-tocopherol and folate appeared adequate. PMID- 16254581 TI - Impact of micronutrient dietary intake and status on intestinal zinc absorption in late middle-aged men: the ZENITH study. AB - BACKGROUND: Adjustments in intestinal absorption and losses of zinc (Zn) are thought to maintain Zn homeostasis when dietary intake levels are altered. Zn status may also influence efficiency of intestinal Zn absorption. OBJECTIVES: To determine the impact of dietary intake and status of some micronutrients on Zn absorption in late middle-aged men. DESIGN AND PARTICIPANTS: Dietary intake and status of Zn, Cu, Fe, vitamin A, C and fibre, and absorption of Zn were measured in 48 men, aged 58-68 y, confined to a metabolic unit and consuming a typical French diet. Dietary intake was estimated using 4-day food-intake records (including the weekend) and the GENI program. To assess Zn status, serum, erythrocyte, urine Zn levels and serum alkaline phosphatase activity were determined. Zn absorption was determined using the isotope double-labelling method. Zn stable isotopic ratios were measured in plasma samples collected before and 48 h after isotope administration using ICP/MS. RESULTS: Zn intake within the group of men varied from 5.7 to 20.5 mg/day and averaged 12.9 mg/day. Serum Zn level varied from 10 to 18 micromol/l and averaged 12.9 micromol/l. Zn absorption varied from 12 to 46% and averaged 29.7%. Zn absorption was not significantly (P > 0.05) correlated with Zn intake or with any of the Zn status parameters. Zn absorption was only slightly negatively correlated with serum and erythrocyte Zn levels and with serum Fe and ferritin levels in this study. CONCLUSION: Zn dietary intake and Zn absorption were satisfactory and led to an adequate Zn status in this population. PMID- 16254582 TI - Introduction to the ZENITH study and summary of baseline results. AB - Zinc is known to be essential for a great number of biochemical activities and physiological and cognitive functions. The objective of Zenith study was to investigate the effects of Zn, as a nutritional supplement, on psychological and behavioural factors and on surrogate markers that are indicative of trends towards better health in order to evaluate the need for dietary recommendations specific to the studied population. We report in this paper the summary of baseline results obtained before Zn supplementation. PMID- 16254583 TI - Basal metabolic rate and thyroid hormones of late-middle-aged and older human subjects: the ZENITH study. AB - OBJECTIVE: This paper describes baseline data on basal metabolic rate (BMR), thyroid hormone levels and body composition of middle-aged and older people participating in the ZENITH project and the correlation of thyroid hormone levels with zinc status. DESIGN: A multicentre prospective intervention study employing a randomised double blind design. SETTING: Clermont-Ferrand, Theix (France), Coleraine (Northern Ireland), Grenoble (France), Rome (Italy). INTERVENTIONS: BMR has been measured on a subsample of 70 middle-aged volunteers (35 men and 35 women recruited in Clermont-Ferrand, France, aged 55-70 y) and 108 older volunteers (56 men and 52 women recruited in Rome, Italy, aged 70-85 y). Thyroid hormone levels were evaluated in the entire group of ZENITH volunteers (n = 387). BMR was measured by indirect calorimetry. Fat-free mass (FFM) was derived by four skinfold thicknesses using Durnin and Womersley's equations. Concentrations of thyroid hormones (total T3 and T4) were measured using a competitive immunoassay with an enhanced chemiluminescence end point. RESULTS: Italian older volunteers had a significantly lower FFM than middle-aged French volunteers (-7% P < 0.01). A negative correlation between BMR and age (men, r = -0.64; women, r = -0.62; both P < 0.0001) was observed: BMR was significantly (P < 0.000001) lower in Italian elderly volunteers (4.03+/-0.46 kJ/min and 3.29+/-0.42 kJ/min for men and women, respectively) than in middle-aged French volunteers (4.84+/-0.45 kJ/min and 3.87+/-0.38 kJ/min for men and women, respectively), even after adjustment for FFM (-12%). No correlation has been observed between BMR and thyroid hormones both in French and Italian subjects. Total T4 (TT4) concentrations were lowest in middle-aged population (-10%, P < 0.0001). A moderate negative correlation has been found with TT4 and red blood cell zinc (r = -0.12, P < 0.02, slope -0.026). CONCLUSIONS: The results confirm an age-related decline in BMR not entirely explained by body composition or thyroid hormones differences. PMID- 16254584 TI - Age-related oxidative stress and antioxidant parameters in middle-aged and older European subjects: the ZENITH study. AB - OBJECTIVE: Oxidative stress has been reported to increase with ageing. However, the data in healthy humans remain controversial and studies in free-living elderly people are scarce. The objective of the present study was to compare age related oxidative stress in late middle-aged and older free-living subjects. DESIGN: The effect of ageing on oxidative stress and antioxidant parameters was investigated in 188 middle-aged subjects from Clermont-Ferrand (France) and Coleraine (UK), and in 199 older subjects from Grenoble (France) and Roma (Italy). Plasma thiol (SH) groups, define definition (TBAR's) and total glutathione (GSH), define definition (FRAP), and superoxide dismutase (SOD) activity were measured at baseline of the ZENITH study. RESULTS: Plasma SH groups and FRAP and, surprisingly, TBAR's were significantly lower in free-living older subjects compared to younger subjects (P < 0.001, P < 0.001, P < 0.01, respectively), but there was no significant differences in GSH levels. CONCLUSION: European free-living healthy older do not appear to be exposed to an acute oxidative stress. However, the highly significant positive correlation between plasma SH group oxidation or decreased FRAP and ageing is predictive of an increased risk of oxidative stress in older subjects. Moreover, the comparison between middle-aged and older subjects regarding oxidative stress parameters suggests also a progressive and slow decline of antioxidant status in healthy free-living older elderly and underline the impact on life-style factors on successful ageing. PMID- 16254585 TI - Zinc status and age-related changes in peripheral blood leukocyte subpopulations in healthy men and women aged 55-70 y: the ZENITH study. AB - OBJECTIVE: To determine zinc status and age-related changes in the immune function of healthy late-middle-aged men and women (aged 55-70 y). DESIGN: Observational study. SETTING: Population of Northern Ireland. SUBJECTS: Apparently healthy, free-living individuals (45 men, 48 women) aged 55-70 y. INTERVENTION: Zinc status markers were analysed by flame atomic absorption spectrometry and commercially available kits. Immune function was assessed by flow cytometry. RESULTS: Serum and erythrocyte zinc concentrations were 13.0 (s.d. 1.40) micromol/l and 222 (s.d. 48.2) micromol/l, respectively. Serum alkaline phosphatase (ALP) concentrations were 76.8 (s.d. 16.1) U/l; women showed significantly higher concentrations of ALP (P = 0.011). Women demonstrated (1) a significant inverse correlation in naive T lymphocytes, specifically naive T helper lymphocytes (% expression, r = -0.364, P = 0.007 and absolute count, r = 0.275, P = 0.036) with age and (2) a significant positive correlation between late activation of T lymphocytes (% expression, r = 0.299, P = 0.019 and absolute count, r = 0.260, P = 0.039) with advancing age. Men demonstrated a significant positive correlation in the % expression of (CD3-/CD16+/CD56+) natural killer (NK) cells with age (r = 0.316, P = 0.017). CONCLUSIONS: Between the ages of 55 and 70 y, healthy individuals experience significant alterations in immune function; however, such changes appear largely sex specific. Given the reported importance of adequate zinc status in maintaining optimal immune function, further studies are required to explore the effect of enhanced zinc status on emerging immune deficiencies in cell-mediated immunity in healthy 55-70 y olds. PMID- 16254586 TI - Immune response in relation to zinc status, sex and antioxidant defence in Italian elderly population: the ZENITH study. AB - OBJECTIVE: Evaluation of some immune markers in Italian elderly population in relation to zinc status, gender and antioxidant defence. DESIGN: Observational study. SETTING: Italian population. SUBJECTS: Apparently healthy, free-living subjects, 56 men and 52 women, aged 70-85 y, enrolled in Italy. METHODS: Lymphocytes were unstimulated or stimulated with the mitogen phytohemoagglutinin (PHA). The proliferative capacity was measured as incorporation of [3H]-thymidine and reported as stimulation index (SI). Cytokine secretion by lymphocytes was determined by ELISA. The antioxidant enzyme activities were measured using commercial kits. RESULTS: Dietary zinc intake, as well as zinc in serum, red blood cells and urine were on the normal range of values and did not show any difference between men and women. The proliferative response showed a high variability without significant differences between men and women. The amount of secreted pro- and anti-inflammatory cytokines was similar in men and women. No differences were found in the activity of antioxidant enzymes in lymphocytes, namely superoxide dismutase, glutathione peroxidase and catalase, between men and women. An association between SI and serum zinc level in men was found. SI resulted negatively correlated with interleukin (IL)-1beta (R2 = 0.036 and P = 0.012) and IL-10 (R2 = 0.34 and P = 0.040) only in men. IL-10 of PHA-stimulated lymphocytes was negatively correlated with red blood zinc in men (R2 = 0.41 and P = 0.008), while IL-10 of unstimulated and PHA-stimulated lymphocytes were negatively correlated with serum zinc in women (R2 = 0.38 and P = 0.020; R2 = 0.31 and P = 0.040, respectively). No correlation was observed between immune markers and antioxidant enzyme activities. CONCLUSIONS: Only weak differences on immune response between men and women were observed. However, zinc status appears to have more influence on the ability of lymphocytes to proliferate in men than in women. PMID- 16254587 TI - The relationship between the zinc nutritive status and biochemical markers of bone turnover in older European adults: the ZENITH study. AB - OBJECTIVE: To investigate the relationship between indices of zinc nutritive status and biochemical markers of bone turnover in older adult European subjects. DESIGN: Use of baseline data from a multicentre prospective zinc intervention (ZENITH) study. SETTING: Centres in France, Italy and Northern Ireland. PARTICIPANTS: A total of 387 healthy adults, aged 55-87 y. METHODS: Zinc intake was assessed by 4-day recall records. Circulating and urinary biochemical zinc status measures were assessed by atomic absorption spectrophometry. Serum bone specific alkaline phosphatase and osteocalcin were assessed by ELISA and urinary pyridinoline (Pyr) and deoxypyridinoline (Dpyr) by HPLC. RESULTS: Zinc intake was negatively correlated with urinary Pyr and Dpyr (r = -0.298 and -0.304, respectively; P < 0.0001), but was not correlated with bone formation markers. There was a tendency for serum zinc to be negatively correlated with urinary Dpyr (r = -0.211; P = 0.080). Erythrocyte zinc was negatively correlated with serum osteocalcin (r = -0.090; P < 0.0001). None of the other correlations were significant. After adjustment for confounder (age, gender and research centre) the only significant association that remained was between serum osteocalcin and erythrocyte zinc (beta = -0.124; P = 0.011). CONCLUSIONS: There was some, albeit inconsistent, evidence of a relationship between zinc nutritive status and bone turnover in the older adult participants of the ZENITH study. PMID- 16254588 TI - Screening and recruitment procedure of late-middle aged and older subjects: the ZENITH study. AB - OBJECTIVE: To report the rational, design, recruitment, baseline characteristics and preliminary overview of volunteers in the ZENITH study. DESIGN: A multicentre prospective intervention study employing a randomised double-blind design. SETTING: Clermont-Ferrand, Theix (France), Coleraine (Northern Ireland), Grenoble (France), Rome (Italy). PARTICIPANTS: Healthy men and women middle-aged (55-70 y) and older volunteers (70-87 y). INTERVENTIONS: At baseline (prior to zinc (Zn) supplementation), all volunteers underwent a full clinical examination, anthropometric measurements, health and lifestyle questionnaire, Mini Mental State Examination, Geriatric Depression Scale, and biochemistry profile. RESULTS: In total, 842 volunteers (378 men and 464 women) were invited to take part in the study. A total of 49% of these volunteers were excluded on the basis of inclusion/exclusion criteria. In total, 433 participants were admitted to the Zn supplementation for 6 months. During this period, about 10% of volunteers dropped out from the study. CONCLUSIONS: A total of 387 subjects (197 male and 190 female subjects) successfully completed the supplementation phase of the ZENITH study. PMID- 16254589 TI - Continuous expression of the homeobox gene Pax6 in the ageing human retina. AB - PURPOSE: In the past few years, the essential role of the homeobox gene Pax6 for eye development has been demonstrated unambiguously in a variety of species including humans. In humans, Pax6 mutations lead to a variety of ocular malformations of the anterior and posterior segment. However, little is known about PAX6 expression in the adult human retina. We have therefore investigated PAX6 levels and localization in the human retina at various ages. METHODS: Adult human eyes of various ages (17-79 years) were obtained from the Zurich Eye Bank. PAX6 expression levels and patterns were analysed by Western blot analysis of total retinal protein and by immunohistochemistry on paraffin sections, respectively. RESULTS: PAX6 expression in the retina was detected up to 79 years of donor age and was predominantly localized to the ganglion cell layer and the inner part of the inner nuclear layer. CONCLUSIONS: PAX6 remains distinctly expressed throughout the lifespan of the human retina suggesting a role for PAX6 in the retina after completion of eye morphogenesis. PMID- 16254590 TI - A 6-month assessment of bimatoprost 0.03% vs timolol maleate 0.5%: hypotensive efficacy, macular thickness and flare in ocular-hypertensive and glaucoma patients. AB - AIM: To compare 6 months of treatment with bimatoprost and timolol in terms of their hypotensive efficacy and secondary effects, including changes in macular thickness and the inflammatory reaction induced in the anterior chamber. METHODS: A prospective, randomized, parallel-group trial performed on 30 eyes of 30 patients per group. The main outcome measure was the difference between the IOP value taken between the baseline visit and the 6-month-visit. Macular thickness determined through optical coherence tomography and anterior chamber inflammation estimated using the laser flare meter was also evaluated. Adverse events were recorded during the study period. RESULTS: Bimatoprost treatment gave rise to a significantly lower mean IOP than timolol in all follow-up visits as from the first month (P<0.05). Bimatoprost achieved high percentage IOP reductions from baseline in a significantly higher proportion of patients (P<0.05). Macular thickness and anterior chamber flare failed to vary significantly both between the two groups and within each group during the 6-month evaluation (P>0.05). CONCLUSIONS: Bimatoprost 0.03% once daily showed a greater efficacy then timolol 0.05% twice daily in patients with elevated IOP. No significant differences were detected in macular thickness or anterior uveitis using optical coherence tomography and laser flare photometry. PMID- 16254592 TI - Observer interpretation variability of peripapillary flow using the Heidelberg Retina Flowmeter. AB - PURPOSE: To evaluate the intraobserver and interobserver reproducibility of the automatic full field perfusion image analysis (AFFPIA) program on Heidelberg Retina Flowmeter (HRF) derived perfusion images in a multicentre study group. METHODS: A total of 10 subjects were consecutively recruited in the study. One eye was randomly selected for each patient. Blood flow was assessed by HRF and flow measurements were analyzed by using the AFFPIA program. AFFPIA calculates the Doppler frequency shift and the haemodynamic variables: flow for each pixel. Intraobserver and interobserver reproducibility was calculated for AFFPIA program. The retinal blood flow was calculated in the superior and inferior section, furthermore, each section was divided into three parts: the temporal area, the nasal, and the rim area, as for software, but only the temporal and nasal areas were considered in this study. The blood flow and the area considered were evaluated for each part. RESULTS: When the intraobserver and intraimage reproducibility was studied, the coefficient of variation ranged from 0.4 to 1.9%. When the interobserver and intraimage reproducibility was studied, the retinal blood flow coefficient of variation ranged from 0.52 to 3.30% for the supero-temporal area, from 0.13 to 2.67% for the inferotemporal area, from 0.15 to 2.75% for the supero-nasal area, and from 0.04 to 5.65% for the infero-nasal area. CONCLUSION: Our results with AFFPIA showed an interobserver coefficient of variation of retinal blood flow measurements always less than 6% in both temporal and nasal areas. No significant difference was found among the four observers for the flow measurements. PMID- 16254594 TI - Vitreoretinal surgery under local anaesthesia: missed fellow eye pathology. PMID- 16254595 TI - Posterior polar cataract: minimizing risk of posterior capsule rupture. AB - PURPOSE: To study a preferred technique of phacoemulsification in eyes with posterior polar cataract and report its outcome. METHODS: Under topical anesthesia, phacoemulsification was carried out after hydrodelination in 23 cases (38 eyes) with ages ranging from 19 to 65 years (mean=33.5 years). Hydrodissection was not performed. RESULTS: Mean duration of follow-up was 9.5 months. None of the eyes developed posterior capsule rupture, but seven eyes (18.4%) revealed posterior capsule plaque postoperatively, which needed neodymium : YAG laser capsulotomy. Mean visual acuity improved significantly after surgery (P=0.0001, paired t-test); In all, 34 eyes achieved a best-corrected visual acuity of 20/40 or more (89.4%). However, the postoperative visual acuity was less than 20/25 in 11 eyes (28.9%). The causes of the low acuity were amblyopia in eight eyes (21.0%) and macular degeneration due to retinitis pigmentosa in two others (5.2%). CONCLUSION: Phacoemulsification is an effective and safe method to treat posterior polar cataract with gentle hydrodelination 'hydrodissection free phacoemulsification technique'. This is especially true when great attention is paid to the 'floppy' posterior capsule. Although previous amblyopia might interfere with excellent surgical outcome in patients with a unilateral or highly asymmetric bilateral cataract, visual acuity improved significantly in most cases. PMID- 16254596 TI - Ophthalmic nurse practitioner led diabetic retinopathy screening. Results of a 3 month trial. AB - PURPOSE: To describe the design and implementation of a nurse led diabetic retinopathy screening clinic. To present the results of a 3-month trial period assessing the concordance of retinopathy grading between a nurse practitioner and an ophthalmologist. METHOD: Patients attending for annual diabetic eye review during an initial 3-month trial period were assessed in a dedicated diabetic eye clinic by an ophthalmic nurse practitioner and an ophthalmologist, with both grading the degree of diabetic retinopathy using to the Wisconsin grading system. Each was masked as to the other's findings. The concordance of retinopathy grading between ophthalmic nurse practitioner and ophthalmologist was assessed. RESULTS: A total of 95 patients (189 eyes) were assessed during the study period. A 92% concordance was achieved between the ophthalmologist and the ophthalmic nurse practitioner. In total, 72 eyes were graded as having some degree of retinopathy by the ophthalmologist. The sensitivity of the nurse practitioner for diagnosing the presence of diabetic retinopathy was 93%, and the specificity 91%. Nine eyes with severe nonproliferative diabetic retinopathy or worse, and four with clinically significant macular oedema were seen. All were correctly identified by the nurse practitioner. CONCLUSIONS: The structure and management protocols of the clinic are described. An excellent concordance between ophthalmologist and nurse practitioner was achieved in this group of patients with relatively less advanced retinopathy. PMID- 16254597 TI - Anthrax lethal toxin-mediated killing of human and murine dendritic cells impairs the adaptive immune response. AB - Many pathogens have acquired strategies to combat the immune response. Bacillus anthracis interferes with host defenses by releasing anthrax lethal toxin (LT), which inactivates mitogen-activated protein kinase pathways, rendering dendritic cells (DCs) and T lymphocytes nonresponsive to immune stimulation. However, these cell types are considered resistant to killing by LT. Here we show that LT kills primary human DCs in vitro, and murine DCs in vitro and in vivo. Kinetics of LT mediated killing of murine DCs, as well as cell death pathways induced, were dependent upon genetic background: LT triggered rapid necrosis in BALB/c-derived DCs, and slow apoptosis in C57BL/6-derived DCs. This is consistent with rapid and slow killing of LT-injected BALB/c and C57BL/6 mice, respectively. We present evidence that anthrax LT impairs adaptive immunity by specifically targeting DCs. This may represent an immune-evasion strategy of the bacterium, and contribute to anthrax disease progression. We also established that genetic background determines whether apoptosis or necrosis is induced by LT. Finally, killing of C57BL/6-derived DCs by LT mirrors that of human DCs, suggesting that C57BL/6 DCs represent a better model system for human anthrax than the prototypical BALB/c macrophages. PMID- 16254598 TI - Low effective dispersal of asexual genotypes in heterogeneous landscapes by the endemic pathogen Penicillium marneffei. AB - Long-distance dispersal in microbial eukaryotes has been shown to result in the establishment of populations on continental and global scales. Such "ubiquitous dispersal" has been claimed to be a general feature of microbial eukaryotes, homogenising populations over large scales. However, the unprecedented sampling of opportunistic infectious pathogens created by the global AIDS pandemic has revealed that a number of important species exhibit geographic endemicity despite long-distance migration via aerially dispersed spores. One mechanism that might tend to drive such endemicity in the face of aerial dispersal is the evolution of niche-adapted genotypes when sexual reproduction is rare. Dispersal of such asexual physiological "species" will be restricted when natural habitats are heterogeneous, as a consequence of reduced adaptive variation. Using the HIV associated endemic fungus Penicillium marneffei as our model, we measured the distribution of genetic variation over a variety of spatial scales in two host species, humans and bamboo rats. Our results show that, despite widespread aerial dispersal, isolates of P. marneffei show extensive spatial genetic structure in both host species at local and country-wide scales. We show that the evolution of the P. marneffei genome is overwhelmingly clonal, and that this is perhaps the most asexual fungus yet found. We show that clusters of genotypes are specific to discrete ecological zones and argue that asexuality has led to the evolution of niche-adapted genotypes, and is driving endemicity, by reducing this pathogen's potential to diversify in nature. PMID- 16254599 TI - Open-access science: a necessity for global public health. PMID- 16254600 TI - Cytoskeletal rearrangements in synovial fibroblasts as a novel pathophysiological determinant of modeled rheumatoid arthritis. AB - Rheumatoid arthritis is a chronic inflammatory disease with a high prevalence and substantial socioeconomic burden. Despite intense research efforts, its aetiology and pathogenesis remain poorly understood. To identify novel genes and/or cellular pathways involved in the pathogenesis of the disease, we utilized a well recognized tumour necrosis factor-driven animal model of this disease and performed high-throughput expression profiling with subtractive cDNA libraries and oligonucleotide microarray hybridizations, coupled with independent statistical analysis. This twin approach was validated by a number of different methods in other animal models of arthritis as well as in human patient samples, thus creating a unique list of disease modifiers of potential therapeutic value. Importantly, and through the integration of genetic linkage analysis and Gene Ontology-assisted functional discovery, we identified the gelsolin-driven synovial fibroblast cytoskeletal rearrangements as a novel pathophysiological determinant of the disease. PMID- 16254601 TI - The axon guidance receptor gene ROBO1 is a candidate gene for developmental dyslexia. AB - Dyslexia, or specific reading disability, is the most common learning disorder with a complex, partially genetic basis, but its biochemical mechanisms remain poorly understood. A locus on Chromosome 3, DYX5, has been linked to dyslexia in one large family and speech-sound disorder in a subset of small families. We found that the axon guidance receptor gene ROBO1, orthologous to the Drosophila roundabout gene, is disrupted by a chromosome translocation in a dyslexic individual. In a large pedigree with 21 dyslexic individuals genetically linked to a specific haplotype of ROBO1 (not found in any other chromosomes in our samples), the expression of ROBO1 from this haplotype was absent or attenuated in affected individuals. Sequencing of ROBO1 in apes revealed multiple coding differences, and the selection pressure was significantly different between the human, chimpanzee, and gorilla branch as compared to orangutan. We also identified novel exons and splice variants of ROBO1 that may explain the apparent phenotypic differences between human and mouse in heterozygous loss of ROBO1. We conclude that dyslexia may be caused by partial haplo-insufficiency for ROBO1 in rare families. Thus, our data suggest that a slight disturbance in neuronal axon crossing across the midline between brain hemispheres, dendrite guidance, or another function of ROBO1 may manifest as a specific reading disability in humans. PMID- 16254602 TI - Gli2 and Gli3 localize to cilia and require the intraflagellar transport protein polaris for processing and function. AB - Intraflagellar transport (IFT) proteins are essential for cilia assembly and have recently been associated with a number of developmental processes, such as left right axis specification and limb and neural tube patterning. Genetic studies indicate that IFT proteins are required for Sonic hedgehog (Shh) signaling downstream of the Smoothened and Patched membrane proteins but upstream of the Glioma (Gli) transcription factors. However, the role that IFT proteins play in transduction of Shh signaling and the importance of cilia in this process remain unknown. Here we provide insights into the mechanism by which defects in an IFT protein, Tg737/Polaris, affect Shh signaling in the murine limb bud. Our data show that loss of Tg737 results in altered Gli3 processing that abrogates Gli3 mediated repression of Gli1 transcriptional activity. In contrast to the conclusions drawn from genetic analysis, the activity of Gli1 and truncated forms of Gli3 (Gli3R) are unaffected in Tg737 mutants at the molecular level, indicating that Tg737/Polaris is differentially involved in specific activities of the Gli proteins. Most important, a negative regulator of Shh signaling, Suppressor of fused, and the three full-length Gli transcription factors localize to the distal tip of cilia in addition to the nucleus. Thus, our data support a model where cilia have a direct role in Gli processing and Shh signal transduction. PMID- 16254603 TI - Perspectives on human genetic variation from the HapMap Project. AB - The completion of the International HapMap Project marks the start of a new phase in human genetics. The aim of the project was to provide a resource that facilitates the design of efficient genome-wide association studies, through characterising patterns of genetic variation and linkage disequilibrium in a sample of 270 individuals across four geographical populations. In total, over one million SNPs have been typed across these genomes, providing an unprecedented view of human genetic diversity. In this review we focus on what the HapMap Project has taught us about the structure of human genetic variation and the fundamental molecular and evolutionary processes that shape it. PMID- 16254604 TI - Gain-of-function screen for genes that affect Drosophila muscle pattern formation. AB - This article reports the production of an EP-element insertion library with more than 3,700 unique target sites within the Drosophila melanogaster genome and its use to systematically identify genes that affect embryonic muscle pattern formation. We designed a UAS/GAL4 system to drive GAL4-responsive expression of the EP-targeted genes in developing apodeme cells to which migrating myotubes finally attach and in an intrasegmental pattern of cells that serve myotubes as a migration substrate on their way towards the apodemes. The results suggest that misexpression of more than 1.5% of the Drosophila genes can interfere with proper myotube guidance and/or muscle attachment. In addition to factors already known to participate in these processes, we identified a number of enzymes that participate in the synthesis or modification of protein carbohydrate side chains and in Ubiquitin modifications and/or the Ubiquitin-dependent degradation of proteins, suggesting that these processes are relevant for muscle pattern formation. PMID- 16254606 TI - Heat shock protein HSP70 increases the resistance of cortical cells to glutamate excitotoxicity. AB - Preincubation of cultured slices of the olfactory cortex of rat brain with heat shock protein in a concentration of 1 microg/ml protected the pre- and postsynaptic mechanisms of glutamatergic synaptic transmission from glutamate excitotoxicity (50 mM) inducing blockade of excitatory postsynaptic function and reducing presynaptic processes. It was hypothesized that heat shock protein protects AMPA and NMDA receptor-mediated processes. PMID- 16254605 TI - Discovery of human inversion polymorphisms by comparative analysis of human and chimpanzee DNA sequence assemblies. AB - With a draft genome-sequence assembly for the chimpanzee available, it is now possible to perform genome-wide analyses to identify, at a submicroscopic level, structural rearrangements that have occurred between chimpanzees and humans. The goal of this study was to investigate chromosomal regions that are inverted between the chimpanzee and human genomes. Using the net alignments for the builds of the human and chimpanzee genome assemblies, we identified a total of 1,576 putative regions of inverted orientation, covering more than 154 mega-bases of DNA. The DNA segments are distributed throughout the genome and range from 23 base pairs to 62 mega-bases in length. For the 66 inversions more than 25 kilobases (kb) in length, 75% were flanked on one or both sides by (often unrelated) segmental duplications. Using PCR and fluorescence in situ hybridization we experimentally validated 23 of 27 (85%) semi-randomly chosen regions; the largest novel inversion confirmed was 4.3 mega-bases at human Chromosome 7p14. Gorilla was used as an out-group to assign ancestral status to the variants. All experimentally validated inversion regions were then assayed against a panel of human samples and three of the 23 (13%) regions were found to be polymorphic in the human genome. These polymorphic inversions include 730 kb (at 7p22), 13 kb (at 7q11), and 1 kb (at 16q24) fragments with a 5%, 30%, and 48% minor allele frequency, respectively. Our results suggest that inversions are an important source of variation in primate genome evolution. The finding of at least three novel inversion polymorphisms in humans indicates this type of structural variation may be a more common feature of our genome than previously realized. PMID- 16254607 TI - Blockade of beta-adrenoceptors and muscarinic cholinergic receptors modulates effect on nitric oxide on heart rate in rats. AB - Nitroglycerine in doses of 0.4-1.0 mg/kg decreased the heart rate in rats, which was associated with inhibition of adrenergic influences realized via beta adrenoceptors. The negative chronotropic effect of sodium nitroprusside in a dose of 1 mg/kg was more significant compared to that of nitroglycerine (by 2-3 times). It was associated with inhibition of adrenergic and stimulation of cholinergic influences mediated via beta-adrenoceptors and muscarinic cholinergic receptors, respectively. During blockade of beta-adrenoceptors and muscarinic cholinergic receptors, sodium nitroprusside increased the time of atrioventricular conduction. These data indicate that function of myocytes in the heart conduction system of rats depends on the PQ interval. PMID- 16254608 TI - Effect of opioid antagonist naloxone on maternal motivation in albino rats. AB - We studied the effect of nonselective antagonist of opioid receptor naloxone on the behavior of albino female rats on days 4-6 after delivery. Intraperitoneal injection of naloxone (5 mg/kg) significantly stimulated maternal reactions (increased the number of approaches to pups, decreased the latency of their transfer into new location). Intranasal naloxone (1 mg/kg) produced similar changes. Naloxone in intraperitoneal dose of 1 mg/kg and intranasal dose of 0.2 mg/kg virtually did not modify maternal behavior. PMID- 16254609 TI - Role of adrenergic mechanisms of erythropoiesis regulation during severe hypoxia. AB - We studied the role of beta-adrenergic mechanisms of regulation of erythropoiesis in the formation of the erythron system reaction during severe hypoxia. Blockade of beta-adrenoceptors after the incidence of hypoxic encephalopathy of different genesis was followed by an increase in the number of committed precursors in the bone marrow, hyperplasia of the erythroid hemopoietic stem, and rise in the count of peripheral blood erythrocytes. These changes were accompanied by decreased formation of abnormal erythrocytes. PMID- 16254610 TI - Disorders in the lungs-placenta system under conditions of experimental gestosis in rats. AB - Study of the arteriovenous difference in hormone levels and hemostasis parameters in rats with experimental gestosis induced by hyper-sodium diet showed decreased production of progesterone, increased level of hydrocortisone (resultant from its increased production and additional release of the hormone by the lungs), hypercoagulation, and retarded fetal development. Involvement of the lungs into the maintenance of optimum rheological parameters of arterial blood and a relationship between the level of fetoplacental hormones and the function of pulmonary fibrinolytic filter were detected. PMID- 16254611 TI - Effect of combined treatment with aspirin and dipyridamole on oxidative homeostasis in mouse serum. AB - Using the method of peroxidative luminol-dependent chemiluminescence we showed that combined treatment with aspirin in low dose and dipyridamole corrects imbalance in oxidative homeostasis in mouse serum. This state resulted from a sharp increase in prooxidant processes after platelet disaggregation with normal dose of aspirin or dose-dependent inhibition of free radical processes with dipyridamole. PMID- 16254613 TI - Circadian dynamics of monocyte phagocytic activity in women during lactation complicated by iron deficiency. AB - Biorhythms of phagocytic activity of peripheral blood monocytes were studied during lactation in healthy women and in women with iron deficiency. Circadian organization of monocyte function was characteristic of healthy nursing women. Strain in the system was associated with elevation of the mean circadian values for reserve potential of the absorption and digestive capacity of phagocytes. Circadian rhythms of the phagocytic and digestive capacity of peripheral blood monocytes were leveled during lactation complicated by iron deficiency. Decreased coefficient of the parameters activation is an early manifestation of latent iron deficiency. Decrease in the basic function parameters and reserve potential are directly related to the decrease in serum ferritin concentration. PMID- 16254612 TI - Function of cytoplasmic NAD-dependent malate dehydrogenase from rat myocardium under conditions of ischemia. AB - NAD-dependent malate dehydrogenase activity decreased by 2.7 times in the myocardium of rats with experimental ischemia. Cytoplasmic NAD-dependent malate dehydrogenase from intact and ischemic rat heart was purified by 91.4 and 95.5 times. We compared kinetic characteristics and regulation of enzyme activity by Fe(2+), Cu(2+), Ca(2+), hydrogen peroxide, and glutathione under normal and pathological conditions. PMID- 16254614 TI - Homocysteinic acid causes oxidative stress in lymphocytes by potentiating toxic effect of NMDA. AB - Short-term incubation of lymphocytes with homocysteine or its oxidation product homocysteinic acid increased the formation of reactive oxygen species and cell necrosis (in case of homocysteinic acid). Effective concentration of homocysteine and homocysteinic acid (500 microM) significantly surpassed the level observed during hyperhomocysteinemia. The addition of homocysteinic acid in a nontoxic concentration of 100 microM potentiated the toxic effect of NMDA and led to massive cell death. During hyperhomocysteinemia the amount of these metabolites in the blood was much higher than in the brain. Oxidative stress produced by these substances can result from activation of NMDA glutamate receptors that were recently detected on lymphocytes. PMID- 16254615 TI - Kinetics of fatty acid oxidation in low density lipoproteins evaluated by registration of the oxidizer consumption and reaction product yield. AB - Oxidation of arachidonic acid by ROS in vitro can be evaluated by the formation of reaction products (conjugated dienes); this is preceded by a lag period caused by the action of antioxidants (alpha-tocopherol, beta-carotene, and ascorbic acid). In case of ozone titration the oxidizer is consumed even during the lag period, when conjugated dienes are not yet forming. Comparison of the oxidation rate constants for antioxidants, arachidonic and oleic monoenic fatty acids suggests that during the lag period Cu(2+)-initiated forms of O(2) oxidize primarily oleic acid, whose reaction rate constant is much higher than those of antioxidants. Presumably, the duration of lag period during oxidation of arachidonic acid and formation of conjugated dienes is determined also by the content of triglycerides and oleic fatty acid in low density lipoproteins. PMID- 16254616 TI - Oxidative stress in atherosclerosis and diabetes. AB - We measured the content of lipid peroxides in plasma LDL from patients with chronic CHD not accompanied by hypercholesterolemia; CHD and hypercholesterolemia; type 2 diabetes mellitus and decompensation of carbohydrate metabolism; and CHD, circulatory insufficiency, and type 2 diabetes mellitus (without hypercholesterolemia). The content of lipid peroxides in LDL isolated from blood plasma by differential ultracentrifugation in a density gradient was estimated by a highly specific method with modifications (reagent Fe(2+) xylene orange and triphenylphosphine as a reducing agent for organic peroxides). The content of lipid peroxides in LDL from patients was much higher than in controls (patients without coronary heart disease and diabetes). Hypercholesterolemia and diabetes can be considered as factors promoting LDL oxidation in vivo. Our results suggest that stimulation of lipid peroxidation in low-density lipoproteins during hypercholesterolemia and diabetes is associated with strong autooxidation of cholesterol and glucose during oxidative and carbonyl (aldehyde) stress, respectively. These data illustrate a possible mechanism of the progression of atherosclerosis in patients with diabetes mellitus. PMID- 16254617 TI - Ca(2+) transport into sarcoplasmic reticulum and immediate-early response proteins in the myocardium of rats resuscitated after systemic circulatory arrest. AB - Activity of antioxidant defense enzymes and content of stress protein HSP70 in the heart increased in passive and, to a lesser extent, in active rats on day 7 of the postresuscitation period after systemic circulatory arrest. The resistance of membrane structures in the heart to endogenous damaging factors in passive rats was lower than in active animals. The degree of compensation in active rats was much higher than in passive animals at these terms of the postresuscitation period. PMID- 16254618 TI - Effect of kindling on GABAergic neurons of the hippocampus and pyriform cortex. AB - GABAergic neurons in different fields of the hippocampus and pyriform cortex were examined 2 weeks and 1 month after electric stimulation of the ventral hippocampus. The counts of GABAergic neurons in the studied structures decreased significantly. The most pronounced shifts in the pyriform cortex were found in the central compartment. Decreased number of inhibitory elements in the two major epileptogenic structures attests to appreciable restructuring in the functions of their neuronal circles. PMID- 16254619 TI - Desialylation decreases the resistance of apo B-containing lipoproteins to aggregation and increases their atherogenic potential. AB - Subfractions of apo B-containing lipoproteins (VLDL and intermediate-density lipoproteins) with reduced content of sialic acid were found in human blood. These lipoproteins are characterized by high capacity to spontaneous association (aggregation) and stimulated accumulation of cholesterol in smooth muscle cells of human aortic intima. In vitro treatment of apo B-containing lipoproteins with alpha-2,6-sialidase and alpha-2,3-sialidase stimulated aggregation and increased the ability of these particles to potentiate cholesterol accumulation in smooth muscle cells of the intact human aortic intima. Probably, desialylation of various apo B-containing lipoproteins can occur in the blood; this process decreases their resistance to aggregation, and increases the ability of these particles to stimulate accumulation of cholesterol in human aortic intima cells, i.e. increases their atherogenic potential. PMID- 16254621 TI - Interspecies pharmacokinetics of xymedon. AB - Pharmacokinetics of immunomodulator xymedon at different modes of the drug administration was studied in humans, dogs, and rats. the main parameters of xymedon pharmacokinetics varied in different animal species. The results confirmed the efficiency and correctness of using allo-metrical method for extrapolation of pharmacokinetic data for tentative evaluation of drug parameters in humans by the results of preclinical trials on animals. PMID- 16254620 TI - Study of antiparkinsonic activity of panavir on a model of Parkinson syndrome induced by systemic administration of MPTP to outbred rats and C57Bl/6 mice. AB - The effect of panavir on Parkinson's syndrome induced by 1-methyl-4-phenyl 1,2,3,6-tetrahydropyridine was studied in C57Bl/6 mice and outbred albino rats. Two injections of panavir significantly reduced the severity of oligokinesia and autonomic manifestations of experimentally induced Parkinson's syndrome. PMID- 16254622 TI - Natural cytokines: new potentialities of immunotherapy of gastroduodenal ulcer. AB - Addition of splenocytokine therapy to combined therapy of peptic ulcer accelerated healing and prolonged remission. PMID- 16254624 TI - Effect of benzo[a]pyrene on the immune status of mice with anxious-depressive syndrome. AB - We studied the effect of benzo[a]pyrene on cells of lymphoid organs, energy metabolism of blood lymphocytes, and immunological reactivity in mice with anxious depressive syndrome produced by social stress. Benzo[a]pyrene exhibited a more pronounced immunotoxic activity in anxious-depressive animals, which was more than a simple sum of the effects produced by adverse ecological and psychic factors. PMID- 16254623 TI - Counts of stromal precursor cells in heterotopic bone marrow transplants in mice immunized with group a streptococcus antigens. AB - The count of stromal precursor cells in bone marrow transplants from CBA mice, transplanted to animals immunized with killed type 5 group A streptococcus vaccine, decreased 4.5-6.5 times (depending on the transplant age) in comparison with the grafts transplanted to normal recipients. The counts of stromal precursor cells in 1.5-3-month bone marrow transplants from animals immunized with killed streptococcal vaccine transplanted to normal mice were virtually the same, while in 7-month transplants they decreased 2-fold in comparison with their counts in bone marrow transplants from normal CBA mice transplanted to normal animals. The content of stromal precursor cells in the femoral bone marrow of animals immunized with killed streptococcal vaccine was appreciably (3.5 times) higher than in the bone marrow of normal mice. The results attest to an appreciable effect of streptococcal antigens on the bone marrow stromal tissue and suggest that not all stromal precursor cells, whose count increases after injection of antigens, are responsible for transplantability of the stromal tissue in case of its heterotopic transplantation. PMID- 16254625 TI - Mechanism underlying the effect of myelopeptides on lymphocyte proliferation in vitro. AB - We studied the role of monocytes in the effect of myelopid and myelopeptides MP 1, MP-3, MP-5, and MP-6 on functional activity of peripheral blood lymphocytes in the reaction of blast transformation. Myelopeptides MP-1, MP-3, and MP-6 suppressed blast transformation of lymphocytes. The effect depended on the presence of monocytes in a cell culture. PMID- 16254626 TI - Blood coagulation initiates respiratory burst in neutrophils. AB - Using the in vitro reaction of nitroblue tetrazolium reduction we showed that blood coagulation stimulates production of reactive oxygen species by human neutrophils. Heparin and chondroitin sulfate produced by thrombin-activated basophils are good candidates for inductors of these processes. Similar activation probably occurs in vivo under the influence of inductors secreted by mast cells. PMID- 16254627 TI - Dexamethasone suppresses human interleukin-5 gene promoter. AB - Synthetic glucocorticoid dexamethasone suppressed interleukin-5 gene expression in PER-117 human T cells at the level of transcription. The conserved lymphokine element 0 in the interleukin-5 gene promoter context served as a target for dexamethasone. PMID- 16254628 TI - Hemogram and myelogram in progressing non-Hodgkin's lymphomas. AB - Parameters of hemogram and myelogram were studied in patients with aggressive and indolent non-Hodgkin's lymphomas: the relationships between the parameters recorded before treatment and during remission or progress 6 months after chemotherapy were studied by multifactorial analysis. The progress of indolent non-Hodgkin's lymphomas was associated with changes caused by tumor infiltration of the bone marrow; lymphocytosis in the myelogram or hemogram was associated with a relative decrease in the count of granulocytic hemopoietic stem cells. A sign associated with the absence of remission in aggressive non-Hodgkin's lymphomas was decreased level of hemoglobin and erythroid cells. Changes in myelogram attesting to anemia and suppressed erythropoiesis before chemotherapy are additional prognostic factors indicating obligatory intensification of chemotherapy for patients with aggressive non-Hodgkin's lymphomas. PMID- 16254629 TI - Content of 8-hydroxy-2-deoxyguanosine in steroid receptor-positive and receptor negative breast cancer cells. AB - The content of DNA damage marker 8-hydroxy-2-deoxyguanosine in 16 receptor negative and 18 receptor-positive human breast neoplasms was measured by immunohistochemical methods. Positive staining was revealed in 81.3 and 50.0% samples of groups 1 and 2, respectively. The effect of arylhydrocarbon receptor agonist beta-naphthoflavone on the content of 8-hydroxy-2-deoxyguanosine and number of estrogen and progesterone receptors was evaluated in MCF-7 breast cancer cells. The degree of genotoxic damage significantly increased 1 h after combined treatment with estradiol and beta-naphthoflavone (in contrast to individual treatment) and remained practically unchanged in the follow-up period. According to the estrogen effect-switching phenomenon, genotoxic damage can contribute to the development of R(-)-breast cancer. PMID- 16254630 TI - Relative distribution densities of cholinergic and adrenoceptor structures in the central part of the sinoatrial node in rat heart. AB - Characteristics of distribution of cholinergic and adrenoceptor structures along the sinoatrial node artery in rat heart were evaluated by autoradiography on semithin sections by determining the density of (3)H-dihydroalprenolol and (3)H quinuclidinyl benzilate binding sites. The relative density of binding sites for (3)H-dihydroalprenolol and (3)H-quinuclidinyl benzilate was minimum in the functional nucleus of the sinoatrial node and asymmetrically increased to maximum values to cranial (sharply) and caudal (smoothly) directions. The relative level of binding for (3)H-dihydroalprenolol in the perinodal atrial myocardium tissue was markedly lower than in the periarterial zone of the central part of the sinoatrial node and comparable to that for (3)H-quinuclidinyl benzilate. PMID- 16254632 TI - Crystallization of components oral fluid in diabetics in case of absence of crystal structures. AB - Crystallization of mixed salivary pools was studied in patients with types 1 and 2 diabetes mellitus. The formation of microcrystals in two types of diabetes varied and could be differentiated by multidimensional analysis. If salivary crystallization was absent, the material could be evaluated by the texture analysis methods. PMID- 16254631 TI - Reparative osteogenesis during transplantation of mesenchymal stem cells. AB - Reparative osteogenesis was studied after xenotransplantation of suspension cell graft from human mesenchymal stem cells. A model of experimental damage to rat femoral diaphysis was developed. The state of animals was satisfactory and non depressed in the early and late postoperation period. We revealed no local pathological reactions and complications. Administration of mesenchymal stem cells into the area of bone defect accelerated and improved regeneration. Unilateral transplantation of the cell graft stimulated regeneration in the contralateral limb due to acceleration of bone tissue maturation. On day 90 after treatment the bone regenerate was completely developed in the area of defect in animals of various groups. The newly formed bone tissue was well integrated into the bone organ. PMID- 16254633 TI - A new method for evaluation of radioactive label transport intensity in the predominant direction between blood and liver. AB - The intensity of (75)Se transport in the predominant direction after intraperitoneal injection of [(75)Se]selenate was compared in 1- and 3-month-old rats receiving common vivarium ration or sucrose diet. The incorporation percent, blood/liver relative radioactivity, and relative radioactivity difference coefficient were evaluated in the blood and liver. The dynamics of label incorporation in the blood of rats fed common diets has two peaks (at 1-3 h and 12-24 h) and a drop at 6 h. Coefficient of difference in 1-month-old rats was characterized by a greater amplitude of fluctuations than in 3-month-old animals. PMID- 16254634 TI - Use of human VEGF(165) gene for therapeutic angiogenesis in coronary patients: first results. AB - The paper presents the first results of therapeutic angiogenesis in clinical cardiosurgery: human VEGF(165) gene transplantation to coronary patients. The use of this therapeutic method is particularly effective in patients with inoperable cardiovascular injuries, i.e. patients with the most severe condition, in whom treatment remains little effective at the modern level of cardiosurgery development. PMID- 16254635 TI - Formation of neuroepithelial structures in culture of neural stem cells from human brain. AB - Dissociated fetal brain cells in a floating culture form clusters and then neurospheres, some of which contain structures shaped as cell "rosettes". The cells in these "rosettes" are arranged radially around the central cavity, in which their apical processes form desmosome-like contacts. Mitotic division of cells in the "rosettes" is associated with migration of the nuclei, similarly to division of neuroepithelial cells in the neural tube during normal embryogenesis. These cells express nestin, a marker of neural stem cells. The cells in "rosettes" found after transplantation have similar characteristics. PMID- 16254636 TI - Peculiarities of proliferation of epidermal cambial cells in mouse skin. AB - The long axis of epidermal cambial cells determining the direction of their division is determined before the beginning of division and is oriented perpendicularly, but not parallel, to the basal membrane, as in other basal cells. As a result, only one of the two newly formed cells adheres to the basal membrane and at the expense of traction forces detaching one cell from the other and elastic force of the basal membrane is formed as a maternal cell and stays in the focus of multiplication. The other cell, turning around under the effect of traction forces perpendicularly to the maternal cell, cardinally changes the direction of its long axis and is polarized parallel to the basal membrane like other cells. This cell becomes the daughter cell, its shape with the "head" and "tail" allows its rapid migration into other rosettes for differentiation. PMID- 16254637 TI - Proliferative activity of adipocytes in adipose tissue tumors. AB - Electron-autoradiographic study of normal and tumor-transformed adipose tissue (common lipoma and destructive lipoma, i.e. infiltrating and degrading lipoma) showed the capacity of adipose tissue cells in lipomas, especially in destructive lipomas, to proliferation and differentiation. In vivo synthesis of DNA in mature adipocytes not observed previously is described. The role of microvascular wall cells as mesenchymal multipotent precursors in the formation of the adipose tissue is discussed. The involvement of the bone marrow mesenchymal stem cell in this process cannot be ruled out. PMID- 16254638 TI - Use of recombinant AdSV40-BetaGal adenoviral construction for monitoring of transplanted cells. AB - Original recombinant adenoviral construction carrying E. coli beta-galactosidase LacZ gene designed by the authors is convenient for labeling and monitoring of bone marrow mesenchymal (stromal) progenitor cells and myocardial and skin fetal cells transplanted in damaged rat tissues (in the perinecrotic zone of the myocardium and onto burnt skin surface) for their reparation. This genetic construction after pre-inactivation of endogenous beta-galactosidase allows to detect transplanted cells in the foci of injury; positive effects of transplantation on tissue reparation processes can be attributed to the presence of transplanted cells. PMID- 16254639 TI - ElastoPHB membrane systems with immobilized bone marrow stromal cells optimize conditions for regeneration of damaged tissue. AB - The effects of autologous bone marrow stromal cells immobilized on ElastoPHB membranes on reparative processes were studied on a model of rat skeletal muscle injury. Bone marrow stromal cells inhibited substitute (sclerosing) regeneration and activated reparative (reconstructive) regeneration of tissues. PMID- 16254640 TI - Mesenchymal stem cells from human bone marrow and adipose tissue: isolation, characterization, and differentiation potentialities. AB - Comparative study of cultured human bone marrow and adipose tissue (lipoaspirate) mesenchymal stem cells was carried out. The main morphological parameters, proliferative activity, expression of surface and intracellular markers of these cells were characterized. Flow cytofluorometry and histological staining showed that both cell types exhibited similar expression of CD105, CD54, CD106, HLA-I markers, were positively stained for vimentin, ASMA, collagen-1, and fibronectin, but not HLA-DR, CD117, and hemopoietic cell markers. The cells underwent differentiation into adipocytes and osteoblasts under appropriate conditions of culturing. Incubation under neuroinductive conditions led to the appearance of a cell population positively stained for type III beta-tubulin (neuronal differentiation marker). PMID- 16254642 TI - Different effects of enhanced and reduced expression of pub gene on the formation of embryoid bodies by cultured embryonic mouse stem cell. AB - The effects of pub gene on proliferation and initial stages of differentiation of embryonic mouse stem cells were studied in vitro. To this end we used enhanced expression of human pub gene (hpub) and suppression of expression of mouse endogenous pub gene with RNA-interference in embryonic stem cells. Proliferative activity of genetically modified polyclonal lines of the embryonic stem cells transfected with plasmids carrying expressing hpub gene or plasmids generating small interference RNA to this gene did not differ from that of the control cells. Inhibition of expression of endogenous pub gene in embryonic stem cells using small interference RNA 2-fold decreased the formation of embryoid bodies, at the same time additional expression of exogenous hpub gene almost 2-fold increased their number in comparison with the control. It was hypothesized that pub gene participates in early stages of differentiation of embryonic stem cells leading to the formation of embryoid bodies. PMID- 16254643 TI - [Profile of gender violence by intimate partners]. AB - OBJECTIVE: To investigate the prevalence and profile of gender violence (physical, psychological, and sexual) perpetrated against women by current or former intimate partners. METHODS: This is a cross-sectional study carried out at a primary healthcare unit in the city of Porto Alegre, Southern Brazil. Our sample comprised 251 women aged 18-49 years who attended the healthcare unit between October and November 2003. Data were collected by means of a questionnaire and double-entered into a electronic spreadsheet. We carried out univariate and bivariate analyses and the chi-square test. RESULTS: The prevalence of the three types of violence were: psychological (55%, 95% CI: 49 61), physical (38%; 95% CI: 32-44), and sexual (8%; 95% CI: 5-11). Variables significantly associated with the three types of violence included woman's age (psychological: p=0.004), woman's schooling (psychological and physical; p=0.012 and 0.023, respectively), partner's schooling (p=0.004, 0.000), social class (p=0.006, 0.000), years with partner (p=0.006, 0.005), partner's occupation (p=0.015, 0.001), number of pregnancies (p=0.018, 0.037), and prevalence of minor psychiatric disorders (p=0.000, 0.000). CONCLUSIONS: The present study found high prevalences of gender violence perpetrated by intimate partners among the users of a primary healthcare unit. Such units play an important role in preventing violence against women. PMID- 16254641 TI - Osteogenic potential of mesenchymal stem cells from bone marrow in situ: role of physicochemical properties of artificial surfaces. AB - Correlation analysis demonstrated the role of inorganic parameters of the surfaces of calcium phosphate materials in the regulation of osteogenic differentiation of mesenchymal precursors. The progenitor stromal cells were isolated from syngeneic bone marrow immobilized in vitro on calcium phosphate surfaces with different structure, phasic, and elemental composition. After 45 days of subcutaneous ectopic osteogenesis in BALB/c mice, the tissues grown on these matrixes were characterized histologically. It was found that adhesion of bone marrow cells is the initial stage determining their future proliferation (conduction) over the artificial surface and the area of formed tissue plate. The success of histogenesis depends on surface roughness. The optimal roughness class was 4-5 (Russian State Standards), which enables differentiation of progenitor stromal cells under the specific microenvironmental conditions into the connective and adipose tissue cells. Differentiation of the progenitor cells into the stromal cells producing the hemopoiesis-inducing microenvironment also takes place in the foci of active hemopoiesis. Induction of osteogenic potential of the stromal precursors (osteoinduction) is determined by the ratio between calcium and phosphate atoms in surface coatings. In our experimental system, osteogenic differentiation of stromal mechanocytes was blocked only at Ca/P<0.5. PMID- 16254644 TI - [Scale of psychological violence against adolescents]. AB - OBJECTIVE: To present strategies and results of a cross-cultural adaptation of a psychological violence scale to be used in Brazilian studies. METHODS: The scale of violence, a translated version in Portuguese of the original in English, was applied to a sample of 266 7th and 8th grade and junior high school students of public and private schools of the municipality of Sao Goncalo, Southeastern Brazil. Several types of equivalences were investigated. The semantic equivalence was evaluated in the referential and general meaning of each item. The measurement equivalence was assessed through psychometric properties such as test and retest reliability, construct validity, internal consistency and factorial analysis. Reliability was measured by Kappa and intraclass correlation coefficients and construct validity was evaluated by Pearson's coefficient. RESULTS: The theoretical-conceptual discussions were deemed adequate concerning conceptual and items equivalences. The semantic equivalence was above 60 percentile in the evaluation of the referential and general item meaning. Cronbach's alpha was 0.94, Kappa index agreement was discreet, intraclass correlation coefficient was 0.82 and the factorial analysis with one factor structure had a degree of explanation of 43.5% variance. Construct validity showed significant negative correlation with self-esteem, social support, and a positive correlation with parent violence. CONCLUSIONS: The study results indicate the applicability of the instrument in the Brazilian teenager population. PMID- 16254645 TI - Intra-familial physical violence among Mexican and Egyptian youth. AB - OBJECTIVE: To determine the prevalence of experiencing intra-familial violence among Mexican and Egyptian youth and to describe its associated risk factors. METHODS: Data from questionnaires applied to 12,862 Mexican and 5,662 Egyptian youth, aged 10 to 19, who attended public schools were analyzed. Biviarate and logistic regression analysis were used to determine the relationship between socio-demographics, the experience of intra-familial violence and violence perpetration. RESULTS: The prevalence of having experienced intra-familial violence was comparable across the Mexican and Egyptian populations (14% and 17%, respectively). In Mexico, young men were more likely to have experienced such violence (OR=2.36) than women, whereas in Egypt, young women were at slightly greater risk than young men (OR=1.25). Older age, male gender and urban residence were independent correlates of experiencing intra-familial violence among Mexican youth. For Egyptian adolescents, in contrast, younger age, female gender and having non-married parents were independent correlates of victimization. Intra familial violence victims were also more likely than non-victims to perpetrate violence (Mexico: OR=13.13; Egypt: OR=6.58). CONCLUSIONS: Mexican and Egyptian youth experienced intra-familial violence at a relatively low prevalence when compared with youth of other countries. A strong association was found between experiencing intra-familial violence and perpetrating violence. PMID- 16254646 TI - [Parental beliefs and child-rearing attitudes and mental health problems among schoolchildren]. AB - OBJECTIVE: To verify the prevalence and identify the risk factors related to mental health problems among schoolchildren and its possible association with the beliefs and educational attitudes of parents/caretakers. METHODS: Cross-sectional study with a stratified probabilistic sample (n=454) of first to third-graders from public and private schools in Southeastern Brazil. Standardized instruments were administered to parents/caretakers by trained interviewers, including screening questionnaires for mental health problems among children and parents/caretakers; a questionnaire on beliefs and attitudes; and a questionnaire for socio-economic status. Chi-square tests and logistic regression models were used for statistical analysis. RESULTS: We found 35.2% prevalence of clinical/borderline cases among students. Parents/caretakers that believed in corporal punishment as a child-rearing method used physical aggression towards their children more frequently (64.8%). Logistic regression models showed that the act of hitting the child with a belt was associated to conduct problems and to overall mental health problems among schoolchildren in the presence of other risk factors: child gender (male), parents/caretakers with mental health problems, and adverse socioeconomic conditions. CONCLUSIONS: The high prevalence of mental health problems among schoolchildren and its association with child rearing methods and mental health problems among parents/caretakers indicate the need for psycho-educational interventions aimed to reduce physical abuse and mental health problems in childhood. PMID- 16254647 TI - Living conditions and receptive vocabulary of children aged two to five years. AB - OBJECTIVE: To assess the receptive vocabulary of children aged between two years and six months and five years and eleven months who were attending childcare centers and kindergarten schools. METHODS: An analytical cross-sectional study was carried out in the municipality of Embu, Southeastern Brazil. The Peabody Picture Vocabulary Test and analysis of factors associated with children's performance were applied. The sample consisted of 201 children of both genders, aged between two and six years. Statistical analysis was performed using multivariate analysis and logistic regression model. The dependent variable analyzed was test performance and the independent variables were child's age, mother's level of education and family socio-demographic characteristics. RESULTS: It was observed that 44.3% of the children had performances in the test that were below what would be expected for their age. The factors associated with the best performances in the test were child's age (OR=2.4; 95% CI: 1.6-3.5) and mother's education level (OR= 3.2; 95% CI: 1.3-7.4). CONCLUSIONS: Mother's education level is important for child's language development. Settings such as childcare and kindergarten schools are protective factors for child development in families of low income and education. PMID- 16254648 TI - [Prognosis for patients with unilateral Wilms' tumor in Rio de Janeiro, Brazil, 1990-2000]. AB - OBJECTIVE: To analyze the survival and the main prognostic factors among patients with unilateral Wilms' tumor patients. METHODS: The study cohort included 132 patients with unilateral Wilms' tumor aged under 15 years, who were enrolled in a pediatric oncology service. Survival curves were calculated using the Kaplan Meier method and the prognostic factors were analyzed using the Cox proportional hazards model. RESULTS: The overall survival rate for five years was 84.6%. The survival probabilities for disease in stages I, II, III and IV stages were: 100%, 94.2%, 83.2% and 31.3%, respectively. The survival rate was 89.4% for patients with favorable histology, 66.7% for focal anaplasia and 40% for diffuse anaplasia. All patients with stage IV disease and diffuse anaplasia died (n=4). All patients with stage I disease, regardless of histology, remained alive at the end of the follow-up period. CONCLUSIONS: Among the variables selected for the final model, only the staging and the histology remained associated with high risk of death risk, while patients aged 24 - 47 months presented better prognosis than the other patients. These results showed the importance of establishing the diagnosis at an early stage, and that the histology is fundamental for guiding the appropriate therapy. PMID- 16254649 TI - [Prevalence of a set of risk factors for chronic diseases in the city of Sao Paulo, Brazil]. AB - OBJECTIVE: To estimate the prevalence of a set of risk factors for non transmissible chronic diseases and compare it to that found 15-16 years ago in a similar survey. METHODS: A cross-sectional household survey was carried out comprising a random sample of people aged 15-59 years in the city of Sao Paulo between 2001 and 2002. The total of 2,103 people answered a questionnaire and had their blood pressure, weight, height, waist and hip circumferences measured. For a third of these participants, their total cholesterol, HDL-cholesterol, triglycerides and glucose levels were determined. RESULTS: The total age-adjusted prevalences in the study age group were as follows: smoking, 22.6%; uncontrolled blood pressure, 24.3%; obesity, 13.7%; increased waist circumference, 19.7%; total cholesterol >or =240 mg/dL, 8.1%; HDL-cholesterol <40 mg/dL, 27.1%; triglycerides > or =200 mg/dL, 14.4%; and blood glucose > or =110 mg/dL, 6.8%. Smoking, uncontrolled blood pressure, high total cholesterol, low HDL-cholesterol and high triglycerides were significantly more prevalent in men than women. CONCLUSIONS: The prevalences of a set of risk factors for chronic diseases showed men to have a poorer condition than women. In comparison to the previous survey, the prevalence of uncontrolled blood pressure remained unchanged but the prevalence of smoking has significantly lowered. PMID- 16254650 TI - Factors associated to smoking habit among older adults (The Bambui Health and Aging Study). AB - OBJECTIVE: To describe the characteristics and associated factors of the smoking habit among older adults. METHODS: A population-based study was carried out comprising 1,606 (92.2%) older adults (> or =60 years old) living in the Bambui town, Southeastern Brazil in 1997. Data was obtained by means of interview and socio-demographic factors, health status, physical functioning, use of healthcare services and medication were considered. The multiple multinomial logistic regression was used to assess independent associations between smoking habits (current and former smokers) and the exploratory variables. RESULTS: The prevalence of current and past smoking was 31.4% and 40.2% among men, and 10.3% and 11.2% among women, respectively (p<0.001). Among current smokers, men consumed a larger number of cigarettes per day and started the habit earlier than women. Among men, current smoking presented independent and negative association with age (> or =80 years) and schooling (> or =8 years) and positive association with poor health perception and not being married. Among women, independent and negative associations with current smoking were observed for age (75-79 and > or =80 years) and schooling (4-7 and > or =8 years). CONCLUSIONS: Smoking was a public health concern among older adults in the studied community, particularly for men. Yet, in a low schooling population, a slightly higher level was a protective factor against smoking for both men and women. Programs for reducing smoking in the elderly population should take these findings into consideration. PMID- 16254651 TI - [Epidemiological and clinical outlook of chronic Chagas' heart disease in Mexico]. AB - OBJECTIVE: To compare the epidemiological and clinical characteristics of chronic Chagas' heart disease to other dilated cardiomyopathies. METHODS: A study comprising 128 patients from a heart disease center was carried out from 1993 to 2003. Of them, 51 (40%) were Trypasonoma cruzi positive. Epidemiological data was obtained through interviews and clinical and serological data from health services. Statistic analysis was conducted using the Chi-square, Fischer, Mann Whitney or Students' t-test as well as multivariate analysis. RESULTS: Chronic Chagas' disease patients were older (55+/-10 years old) than those patients with cardiopathy (42+/-17 years old). Most of them were born in rural areas (90% vs 68%), lived in poor (75% vs 16%), crowded households (45% vs 20%), together with domestic animals (71% vs 61%) and were aware of the Chagas' vector (73% vs 25%). Rhythm and conduction ECG abnormalities as well as permanent pacemaker were common among Chagas' patients (84% vs 55%, 78% vs 64% and 24% vs 10%, respectively). Congestive heart failure was more frequent among cardiomiopathy patients (88% vs 71%). Both groups had abnormal myocardial perfusion with normal epicardial arteries. Comorbidities were more frequent in cardiomiopathy patients than in chronic Chagas' disease patient (two cases only). CONCLUSIONS: Chagas' disease is the most common cause of dilated cardiomiopathy in the study hospital. Due to its regional distribution in Mexico, it deserves special attention and control programs proven to be effective in other countries. PMID- 16254652 TI - [Social inequalities in cesarean section rates in primiparae, Southern Brazil]. AB - OBJECTIVE: To investigate the effect of social inequalities in cesarean section rates among primiparae having single pregnancy and delivering in maternity hospitals. METHODS: The study was carried out in Southern Brazil in 1996, 1998 and 2000. Data from the Live Birth National Information System were used to estimate annual rates and crude and adjusted odds ratios (OR) of cesarean sections according to social conditions (maternal age and education, newborn skin color/ethnicity and macro-regions), duration of pregnancy, and number of prenatal visits. RESULTS: The overall cesarean section rate was 45%, and above 37% in all macro-regions. Increased rates were seen among native and black mothers, aged 30 years or more, living in metropolitan, river valley and mountain macro-regions and having attended to more than six prenatal visits. Crude and adjusted OR show that cesarean rates were negatively associated with all categories of skin color/ethnicity when compared to white newborns, particularly those of native Brazilian (ORadj=0.43; 95% CI: 0.31-0.59), and they were positively associated with higher maternal education (ORadj=3.52; 95% CI: 3.11-3.99), older age (ORadj=6.87; 95% CI: 5.90-8.00) and greater number of prenatal visits (ORadj=2.16; 95% CI: 1.99-2.35). The effects of age and education were partly mediated by the greater number of prenatal visits among higher educated older women. The OR varied among macro-regions but were greater for the wealthier mountain region. CONCLUSIONS: High rates of cesarean section rates in Southern Brazil are a public health concern. They are associated with social, economic and cultural factors which can lead to misuse of medical technology during labor and delivery. PMID- 16254653 TI - [Prenatal care within Brazil's Unified Health System]. AB - OBJECTIVE: To assess antenatal care in health care units, so as to obtain a baseline for future evaluation studies. METHODS: A self-applied inquiry was conducted among municipal health managers within a probabilistic stratified random sample of 627 municipalities which, through expansion technique, extended the analysis to 5,507 municipalities. Data was collected from October 2003 to April 2004. The survey appraised information about the priority granted by the managers to each modality of care, as well as data concerning characteristics of the assistance provided and the declared estimate of the demand being covered. The Chi-square test and Student's t-test were performed in order to verify independence among the qualitative variables and mean differences, respectively. RESULTS: Almost half (43.8%; n=2,317) of the municipalities did not attend gestational risk; 81% (n=4,277) and 30.1% (n=1,592) reported that they attend over 75% of the demand for low and high risk antenatal respectively; 30.1% (n=1,592) attend over 75% of the demand for high risk care. Care for low risk (chi2=282,080; P<0.001 n=4,277) and for high risk pregnancies (chi2=267.924; P<0.001 n=5,280) were associated to geographic region, municipality's size and management modality within the Unified Health System. The guarantee of vacancy for labour and birth was also associated to management modality. CONCLUSIONS: There were gaps related to the provision and the quality of antenatal care within the Unified Health System. Municipal based health care extends the provision of antenatal care, but there are inequalities among regions and among municipalities according to demographic size. PMID- 16254654 TI - Neonatal mortality in intensive care units of Central Brazil. AB - OBJECTIVE: To identify potential prognostic factors for neonatal mortality among newborns referred to intensive care units. METHODS: A live-birth cohort study was carried out in Goiania, Central Brazil, from November 1999 to October 2000. Linked birth and infant death certificates were used to ascertain the cohort of live born infants. An additional active surveillance system of neonatal-based mortality was implemented. Exposure variables were collected from birth and death certificates. The outcome was survivors (n=713) and deaths (n=162) in all intensive care units in the study period. Cox's proportional hazards model was applied and a Receiver Operating Characteristic curve was used to compare the performance of statistically significant variables in the multivariable model. Adjusted mortality rates by birth weight and 5-min Apgar score were calculated for each intensive care unit. RESULTS: Low birth weight and 5-min Apgar score remained independently associated to death. Birth weight equal to 2,500 g had 0.71 accuracy (95% CI: 0.65-0.77) for predicting neonatal death (sensitivity =72.2%). A wide variation in the mortality rates was found among intensive care units (9.5-48.1%) and two of them remained with significant high mortality rates even after adjusting for birth weight and 5-min Apgar score. CONCLUSIONS: This study corroborates birth weight as a sensitive screening variable in surveillance programs for neonatal death and also to target intensive care units with high mortality rates for implementing preventive actions and interventions during the delivery period. PMID- 16254655 TI - [Maternal periodontal disease as a factor associated with low birth weight]. AB - OBJECTIVE: Recent studies have presented evidence that periodontal disease in pregnant women may be a determining factor for low birth weight. The present investigation was carried out to verify whether or not there is an association between maternal periodontal disease and low birth weight. METHODS: This was a case-control study on 302 women, of whom 102 were the mothers of live newborns of low weight (case group) and 200 were the mothers of live newborns of normal birth weight (control group). The existence of an association between periodontal disease and low birth weight was evaluated by means of a multivariate logistic regression model that considered other risk factors for low weight. RESULTS: The two groups were comparable with regard to age, height, pre-gestational weight, smoking, alcohol use, previous diseases, marital status, socioeconomic status, frequency of tooth-brushing and use of dental floss, number of meals per day and visits to the dentist. Periodontal disease was diagnosed in 57.8% of the mothers in the case group and 39.0% in the control group. Logistic regression analysis indicated a positive association between periodontal disease and low birth weight (unadjusted OR=2.15; 95% CI: 1.32-3.48), especially among the mothers with schooling of less than or equal to four years (ORadjusted=3.98; 95% CI: 1.58 10.10). CONCLUSIONS: Periodontal disease is a possible risk factor for low birth weight. PMID- 16254656 TI - [Reasons why sterilized women refused reversive contraceptive methods]. AB - OBJECTIVE: To identify the representations of contraceptive methods within a group of sterilized women, aiming at understanding the reasons why they refused those methods. METHODS: A descriptive qualitative study was carried out on 31 sterilized women, randomly selected from a list of patients attending the Family Planning Program of a university hospital. Data was collected through a semi structured interview. Interview transcription was analyzed according to the Content Analysis method. RESULTS: Refusal to other contraceptive methods was based on representations formed from a mixture of technical information received at health service units, their previous experiences with contraceptive methods and information received in the social environment. Rejection of high efficacy methods (hormonal and IUD) was mostly based on their low innocuousness; refusal to fertility awareness methods (calendar calculation, Billings ovulation method) was due to their low efficacy; and rejection of barrier methods (condom, diaphragm) was due to a culturally patterned sexuality full of interdictions as well as to their low efficacy. CONCLUSIONS: The option for female sterilization may be indicative of the refusal to the contraceptive alternatives offered by health services. Reproductive health professionals should increase in their practice the knowledge about personal, socio-economic and cultural factors that affect women's choice of a contraceptive method that could guarantee a better control over their own reproductive life. PMID- 16254657 TI - [Professional competence and contraceptive care]. AB - OBJECTIVE: To evaluate the technical competence of professionals carrying out activities related to contraceptive care. METHODS: Evaluative research in the field of health was conducted in eight districts of the State of Ceara from July to September 2003. Data was collected by means of interviews with 29 nurses working in the Family Health Care Program within these districts and 50 people being attended by this program. Observations of the Family Health Care Units were a complementary source of data within this study. RESULTS: The majority of nurses had received some form of training regarding contraception and the technical norms regulating their use. However, professional barriers were reported by the nurses and others were identified by lay persons being attended by the program, that indicate the need to provide better training for professionals engaged in this area of care. The nurses recognized they had deficits in information and communication skills as well as technical deficits in dealing with contraception. CONCLUSIONS: There are gaps in professional competence with regard to contraceptive care that, when associated to the lack of systematization of team work, generates distortions in the quality of care. Team work was characterized by the lack of definition of team members' specific attributions and tasks. PMID- 16254658 TI - Health, sleep and lack of time: relations to domestic and paid work in nurses. AB - OBJECTIVE: To assess whether reported morbidity and complaints of lack of time and sleep are associated with the burden of professional work and housework among nurses. METHODS: A cross-sectional exploratory study was carried out among female nurses and nurse assistants (N=206) of a public hospital in Rio de Janeiro, Brazil. Data were collected by means of a questionnaire. The prevalence ratio and 95% confidence intervals were estimated. RESULTS: Mean duration of professional work and housework time was 40.4 and 31.6 hours/week, respectively. Long professional working time (over 44 hours/week) were associated with mild emotional disorders (PR=1.37; 95% CI: 1.05-1.80), complaints of lack of time for resting/leisure (PR=1.61; 95% CI: 1.31-1.97), housework (PR=1.48; 95% CI: 1.12 1.97), and childcare (PR=1.99; 95% CI: 1.51-2.63). Long housework time (over 28 hours/week) was associated with lower prevalence of lack of time for childcare (PR=0.62; 95% CI: 0.46-0.84). High housework load was associated with lack of personal time and complaints of varicose veins (PR=1.31; 95% CI: 1.14-1.50 and PR=1.31; 95% CI: 1.08-1.58, respectively). Complaints of varicose veins were also frequent among female nurses with a total work load above 84 hours (PR=1.30; 95% CI: 1.05-1.61), though this group has shown a lower prevalence of arterial hypertension and recurrent headaches (PR=0.35; 95% CI: 0.15-0.83 and PR=0.53; 95% CI: 0.32-0.89, respectively). CONCLUSIONS: Results suggest that both professional and home environments are relevant in the evaluation of work overload on nurses' health and their family and social life. It is stressed the need for instruments for analyzing total workload among female populations. PMID- 16254659 TI - [Conceptions and practices of community health agents caring for the elderly]. AB - OBJECTIVE: To assess the main conceptions related to the aging process among community health agents. METHODS: An exploratory descriptive qualitative study was carried out in Pernambuco, Northeastern Brazil, from 2000 to 2002. Data was collected from records and interviews with 148 community health agents. The thematic analysis allowed data classification and assembling. RESULTS: The results show that community health agents identified their main tasks as the development of activities in health education and performance of basic actions. Most of them had negative opinions on the aging process, and complete understanding of the concept of health. They mentioned that most complaints by the elderly are health-related problems and need for affection. Community agents identified caring for the elderly as their main responsibility and inadequate service organization made it difficult to operationalize care delivery. Regarding their expectations, they were found to want to further their knowledge on the aging process. CONCLUSIONS: Community health agents identified themselves as main actors for delivery basic care and core agents in the accomplishment of specific health policies. Their conceptions had a positive perspective in relation to their attributions and aging health. The study points out to the need of promoting agents' training to make them able to deal with the multiple aspects of aging. PMID- 16254660 TI - [Alcohol consumption among women: a qualitative analysis]. AB - OBJECTIVE: To assess life histories of alcohol consumption among women in specialized treatment center for alcohol dependency. METHODS: A qualitative study using a life-history approach was carried out in a sample comprising 13 women being treated in an alcohol and drug treatment and research outpatient clinic from May to August 2000. A focused/theme approach of their experience was used. Thirteen semi-structured interviews were performed and recorded for content analysis. RESULTS: The following categories were identified in the interviews transcribed: 1) "Engaging in pleasant activities, as working and leisure, before alcohol abuse and dependency", 2) "Lost of control over alcohol consumption and onset of clinical, social and family problems", 3) "Harm perception motivating seeking out specialized treatment", 4) "Need to recover self-esteem", 5) "Acceptance and respect for specialized treatment" and 6) "Learning to live again: coping with dependency". CONCLUSIONS: Alcohol abuse women need special attention from health providers and their family, mostly to what concerns their emotional aspects and clinical problems and promotion of self-esteem. These actions should allow them to regain their social role aiming at assuring the ongoing recovery process. PMID- 16254661 TI - [Predictive model for cocaine use in prisons in Rio de Janeiro, Brazil]. AB - OBJECTIVE: To identify predictors of and groups vulnerable to cocaine use in prison. METHODS: We selected 376 inmates with history of cocaine use in prison (cases) and 938 inmates with no history of drug use (controls) serving sentences in the Rio de Janeiro State prison system in 1998. The analysis included exposure variables divided into three hierarchical levels: distal, intermediate, and proximal. We performed bivariate analysis using logistic regression and multivariate analysis using hierarchized regression; results are given in odds ratios. RESULTS: Variables associated with cocaine use in prison in the proximal level were use of alcohol and marijuana and duration of imprisonment in years. The effect of social vulnerability variables (distal level) was intermediated by variables in the next levels. Considering only the distal and intermediate levels, use of marijuana prior to imprisonment (OR=4.50; 95% CI: 3.17-6.41) and offence in order to obtain drugs (OR=2.96; 95% CI: 1.79-4.90) showed the strongest association with the outcome. For every additional year spent in prison, the odds of cocaine use increase by 13% (OR=1.13; 95% CI: 1.06-1.21). CONCLUSIONS: Considering the distal and intermediate levels, use of marijuana prior to imprisonment and perpetration of offence in order to obtain drugs were the variables with greatest predictive value. The final model showed alcohol and marijuana use in prison and duration of imprisonment as important predictors of the outcome. The prison environment appears as a factor stimulating drug use. PMID- 16254662 TI - [Impact of legislation on registration of acutely toxic pesticides in Brazil]. AB - OBJECTIVE: To evaluate the impact of the application of the "Pesticides Law" (Law no. 7,802/89) on the toxicological classification profile of registered pesticides in Brazil, from 1990 to 2000. METHODS: Data on registered commercial products in the years 1990 and 2000 were analyzed according to toxicological class, date of registration of the active ingredient (before or after the Law) and usage class (insecticides, fungicides, herbicides and others). Relative Identified Distribution Analysis was utilized for verifying statistical differences between distributions according to toxicological classes. RESULTS: Among the 863 registered commercial products in 2000, 46.6% were already registered before the Pesticide Law. Among the 461 commercial products registered after the Law, 59.2% were derivatives of active ingredients registered before the Law, and 41.4% were classified as highly hazardous (classes I and II of the Brazilian toxicological classification). No significant differences in toxicological classification profile were identified between the group of pesticides derived from active ingredients registered before the Law and the group derived from active ingredients registered after the Law (p<0.0859). CONCLUSIONS: Ten years after the Pesticide Law came into effect, no significant improvement had been observed in the toxicological classification profile of registered pesticides. This was due especially to the continuing presence of products registered before the Law and the registration of new commercial products derived from "old" active ingredients. There was still a high proportion of registrations in the most hazardous classes. It is recommended that compulsory periodic reevaluation of all registered pesticides should be reestablished. PMID- 16254663 TI - [The Internet influence on the academic-scientific public health community]. AB - OBJECTIVE: To investigate the influence of the Internet on the academic scientific activities of the Brazilian public health community. METHODS: This was a descriptive study, centered on the opinions of 237 teachers connected with Brazilian postgraduate programs in public health, at master's and doctoral levels, in 2001. Data were collected by means of a self-administered questionnaire via the Internet and traditional mail. The statistical analysis was done by means of proportions, means and standard deviations. RESULTS: The vast majority of the population (225; 94.9%) said that they used the Internet. Electronic mail (92%) and the web (55.6%) were the resources in greatest daily use. The Internet had significant influence (73.8%) on communications between the teachers, especially for developing collaborative research. The teachers who did not use the Internet (5.1%) gave the reasons that they lacked motivation and time and that it was easy to obtain the material they needed from their colleagues. CONCLUSIONS: The results showed that the Internet has had an influence on the teachers' work and has been affecting the cycle of scientific communication, particularly due to the high speed with which information can be retrieved. There was a tendency to single out communication between the teachers as the feature that has changed most since the coming of the Internet to the Brazilian academic scientific world. PMID- 16254664 TI - Prevalence of Helicobacter pylori infection in Fortaleza, Northeastern Brazil. AB - The prevalence of Helicobacter pylori infection was assessed in a randomly selected sample of individuals from low-income community in Fortaleza, Northeastern Brazil. Overall, 384 out of 610 participants (62.9%) were H. pylori positive. A 47.5% infection rate was found in subjects aged six months to 10 years old, increased to 73.3% in subjects aged 11-20 years and then continued to increase with age reaching up to 87% in those over 60 years old. After this age group, the prevalence decreased slightly. The prevalence of infection increased significantly with age (p<0.0001). PMID- 16254665 TI - [Rickettsial diseases in Brazil and Portugal: occurrence, distribution and diagnosis]. AB - The present study is an update review on the occurrence and diagnosis of rickettsial diseases in Brazil and Portugal, aiming at promoting their epidemiological surveillance in both countries. A literature review was carried out and unpublished data of laboratories and surveillance systems were presented. The results described the occurrence of rickettsial diseases and infections in Brazil and Portugal, including other new and still poorly understood rickettsial infections. Current diagnostic methods were discussed. As in many other countries, rickettsial diseases and infections seem to be an emerging public health problem. Treated as a minor problem for many decades, the interest in these infections has increased in both countries but further studies are needed to establish their role as a public health problem. PMID- 16254666 TI - [Investigation of measles cases in the State of Sao Paulo after monitoring]. PMID- 16254667 TI - [Growing health: resolution establishes hygiene standards for vegetable-based food and beverages]. PMID- 16254670 TI - Different patterns of atherosclerotic remodeling in the thoracic and abdominal aorta. AB - PURPOSE: To investigate the relationship between the vascular diameter and the extent and histologic characteristics of atherosclerosis in the thoracic and abdominal aortas of patients who died of atherosclerotic disease. METHOD: We measured the vascular diameter and evaluated the percentage atrophy of the medial layer of the thoracic and abdominal aortas of 19 patients who died due to atherosclerotic disease. The extent of plaques, calcification, ulceration, thrombosis, and the amount of fat in the plaques were evaluated semiquantitatively. RESULTS: Atherosclerosis was more severe in the abdominal than the thoracic aorta as indicated by the higher sum of the macroscopic scores (P = .02) and the higher percentage atrophy of the medial layer (P < .001). The diameter of the thoracic, but not of the abdominal aorta, correlated with age (r = 0.56; P = .01), plaque score (r = 0.59; P = .008), calcification score (r = 0.749; P < .001), and fat score (r = 0.48; P = .04). Multiple linear regression showed that age (P = .06) and calcification score (P = .001) were the parameters with the strongest association to thoracic aorta diameter. CONCLUSION: There are some differences regarding atherosclerosis in the thoracic compared to the abdominal aorta. Progressive thoracic aorta atherosclerosis is associated with fat deposition in the plaques, inducing arterial dilation. In the abdominal aorta, atherosclerosis can either have a similar evolution or be associated with less fat deposition in the arterial wall, which would result in more rigidity, hindering compensatory arterial enlargement. PMID- 16254671 TI - Vena cava filters in cancer patients: experience with 50 patients. AB - OBJECTIVE: To study the immediate and late results obtained from the implantation of vena cava filters in cancer patients with deep vein thrombosis concomitant with neoplasia. METHODS: This was a retrospective evaluation of 50 patients with an association of cancer and deep venous thrombosis who underwent interruption of the inferior vena cava and the insertion of permanent vena cava filters. The indications for the procedure, filter implantation technique, early and late complications related to the operation, and the clinical evolution were evaluated. RESULTS: The most frequent indication for filter implantation was the contraindication for full anticoagulant treatment (80%). The femoral vein was the preferred access route (86% of the patients). There were no complications related to the surgical procedure. During the follow-up, the following complications were observed: 1 episode of nonfatal pulmonary thromboembolism, 2 cases of occlusion of the inferior vena cava, and 1 case of thrombus retained in the device. Twenty patients (40%) died due to progression of the neoplasm. CONCLUSIONS: Interruption of the inferior cava vein using an endoluminal filter is a procedure with a low rate of complications. It is a safe and efficient measure for preventing pulmonary embolism in cancer patients who have deep vein thrombosis of the lower limbs. PMID- 16254672 TI - Temporal trends of HIV-related risk behavior among Brazilian military conscripts, 1997-2002. AB - PURPOSE: To present selected results of military conscript surveys related to HIV/AIDS, conducted in Brazil, 1997-2002. METHODS: Questionnaires including information on socio-demographic data, sexual behavior practices, sexually transmitted infections-related problems, and use of injecting drugs were completed by 30970 individuals, obtained through a 2-stage sampling. An index of sexual risk behavior was developed to take into account multiplicity of partners and irregularity of condom use. The HIV infection prevalence rate was estimated for 2002. Logistic regression was used to identify the most important determinants of HIV infection. RESULTS: The percentage of regular condom use increased from 38% (1997) to 49% (2002), and the index of sexual risk behavior decreased from 0.98 in 1997 to 0.87, in 2002. The HIV infection prevalence rate was 0.09%, in 2002, which remained unchanged since 1998 Riskier sexual practices among young men with incomplete education and among "men who have sex with men" were found as well as among the participants who reported at least one sexually transmitted infections - related problem. The most important predictor of HIV infection was to be positive for syphilis. CONCLUSIONS: The estimated value of the HIV infection prevalence supports the diagnosis of a concentrated HIV epidemic, in Brazil. Results indicate that particular attention needs to be paid for regional differentials, and for special subgroups, in Brazil. PMID- 16254673 TI - Surgical treatment of traumatic esophageal perforations: analysis of 10 cases. AB - PURPOSE: Traumatic esophageal perforations are infrequent. They represent a surgical dilemma for surgeons, especially if diagnosis is made late. Recently, it has been reported that mortality due to perforation of the esophagus has diminished independently of time of presentation. The experience with traumatic perforations of the esophagus is reviewed to determine morbidity-mortality and how it is affected by time. METHODS: A retrospective clinical review was made of all patients with a diagnosis of traumatic perforation of the esophagus treated by the author. There were 10 patients, all of them male. Average age was 32 years (range 17 to 63). The cause of trauma was gunshot (7), blunt trauma (1) and foreign body (2). Four patients were treated within 24 hours of injury (early treatment). Treatment of 6 patients was delayed 56 to 168 hours after the injury (delayed treatment). RESULTS: Patients treated early underwent primary repair. Delayed treatment included: primary repair (1), T-tube (2), drainage of cervical abscess and pulmonary decortication (2), and conservative treatment (1). There was 1 death in the delayed group (16.6%). One patient in the early treatment group (25%); 4 (66%) in the delayed treatment group had complications. Postoperative stay in the hospital was an average of 20.5 days for the early treatment group and 38 for the late treatment group. CONCLUSIONS: Mortality of traumatic esophageal perforations has diminished significantly. Morbidity, particularly in delayed treatment, is still very high, with multiple operations and prolonged stays in intensive care units and surgical wards, resulting in high hospital costs. The main factor that seems to influence mortality-morbidity of traumatic esophageal perforations is the time of diagnosis. Every effort should be made to diagnose these injuries early. Once diagnosis is made, treatment should be aggressive and expeditious. PMID- 16254674 TI - Measurement of the flexing force of the fingers by a dynamic splint with a dynamometer. AB - PURPOSE AND METHODS: In order to determine forces acting upon an articular joint during hand rehabilitation, a dynamic splint was built and connected to a dynamometer (capable of measuring forces in the range 0 - 600 gf). Through trigonometric calculation, the authors measured the flexing force in the proximal interphalangeal joint of the middle finger at 30 degrees, 45 degrees, 60 degrees, and 90 degrees of flexion. Measurements were obtained in a population of 40 voluntary adults, 20 females and 20 males, This flexing force was correlated with age, sex, and anthropometric measures. RESULTS: Force in the flexing tendon is maximal at the start of flexion, and decreases as the angle of joint flexion increases. A relationship was observed between finger length and the magnitude of the force exerted on the tendon: the longer the finger, the greater the force exherted upon the tendon. Force is greater at all the measured angles, (except 30 degrees) in males and in individuals of higher stature, and bigger arm span. CONCLUSIONS: The flexing force can be effectively measured at all flexing angles, that it correlates with a number of different anthropometric parameters, and that such data are likely to open the way for future studies. PMID- 16254675 TI - Progress testing: evaluation of four years of application in the school of medicine, University of Sao Paulo. AB - BACKGROUND: Progress testing is a longitudinal tool for evaluating knowledge gains during the medical school years. OBJECTIVES: (1) To implement progress testing as a form of routine evaluation; (2) to verify whether cognitive gain is a continuous variable or not; and (3) to evaluate whether there is loss of knowledge relating to basic sciences in the final years of medical school. METHODS: A progress test was applied twice a year to all students from 2001 to 2004. The mean percentage score was calculated for each school year, employing ANOVA with post hoc Bonferroni test evaluation for each test. RESULTS: Progress testing was implemented as a routine procedure over these 4 years. The results suggest a cognitive gain from first to sixth year in all eight tests, as a continuum (P for trend < .0001). Gain was found to be continuous for basic sciences (taught during the first 2 years), clinical sciences (P < .0001), and clerkship rotation (P < .0001). There was no difference between the performance of men and women. CONCLUSION: Progress testing was implemented as a routine, applied twice a year. Data suggest that cognitive gain during medical training appears to be a continuum, even for basic science issues. PMID- 16254676 TI - Endoscopic treatment of vesical and urethral perforations after tension-free vaginal tape (TVT) procedure for female stress urinary incontinence. AB - Stress urinary incontinence is a problem that is prevalent in women, and its treatment with minimally invasive techniques using synthetic materials has increased recently, although the procedure has also brought increased occurrence of specific complications such as vesical and urethral perforations. We describe 11 cases of endoscopic correction of vesical and urethral perforations due to the use of synthetic material for the treatment of stress urinary incontinence. MATERIALS AND METHOD: Eleven patients were treated for complications after undergoing the TVT(R) (tension-free vaginal tape) procedure; 6 of them had the polypropylene tape inside the bladder, and 5 had erosion of the urethra. Endoscopic resection of the polypropylene tapes was performed on all patients. RESULTS: A 6-month follow-up with cystoscopic control showed that the procedures were successful with complete relief of the symptoms except for 1 patient who persisted with the polypropylene tape in the bladder. This patient underwent a new endoscopic resection, and the cystoscopic control exam was normal 3 months later. CONCLUSION: Endoscopic resection of intravesical and intraurethral synthetic tapes can be considered a good alternative for the treatment of complications resulting from the TVT procedure. PMID- 16254677 TI - Second eye cataract surgery: perceptions of a population assisted at a university hospital. AB - OBJECTIVE: To identify daily life difficulties perceived by patients suffering from senile cataract before and after second eye cataract surgery. METHODS: Longitudinal prospective study with 84 patients consecutively seen within the framework of the Cataract Project, with visual acuity equal to or higher than 20/30 in the pseudophakic eye, and equal to or lower than 20/40 with the best possible optical correction in the cataractous eye. A questionnaire was applied during an interview. RESULTS: Before surgery, 60.7% complained about visual impairment (moderate or marked); after surgery, 92.8% had no difficulty. Routine activities, mobility, and leisure activities were significantly altered after surgery (P =.001). CONCLUSION: According to the patients' perceptions, there was a significant reduction in visual difficulties after second eye cataract surgery. PMID- 16254678 TI - Prevention of preeclampsia with low-dose aspirin -- a systematic review and meta analysis of the main randomized controlled trials. AB - The purpose of this paper is to evaluate the effectiveness of low-dose aspirin in the prevention of preeclampsia in low-risk and high-risk women. We identified randomized clinical trials of the use of low-dose aspirin to prevent preeclampsia through the PUBMED search engine, and through the Cochran Library database. Twenty-two studies met our inclusion criteria, and were divided according to the studied population into 2 groups: trials with women at low risk for preeclampsia and trials with women at high risk. Effects were measured through the incidence of preeclampsia in women taking either placebo or aspirin, in studies where the relative risks and the 95% confidence intervals were calculated for both groups. A total of 33,598 women were studied, comprising 5 trials with 16,700 women at low-risk and 17 trials including 16,898 women at high risk. The incidence of preeclampsia was 3.75% (626/17,700), in the low-risk group, 9.01% (1,524/16,898) in the high-risk group, and 6.40% (2,150/33,598) overall. Low-dose aspirin had no statistically significantly effect on the incidence of preeclampsia in the low risk group (RR = 0.95, 95% CI = 0.81-1.11), but had a small beneficial effect in the high-risk group (RR = 0.87, 95% CI = 0.79-0.96). Therefore, low-dose aspirin is mildly beneficial in terms of reducing the incidence of preeclampsia in women at high risk of developing preeclampsia. PMID- 16254679 TI - Noninvasive detection of coronary artery disease -- challenges for prevention of disease and clinical events. AB - Atherosclerosis is a chronic inflammatory disease that affects essentially all arterial beds including the aorta, coronaries, carotids, and peripheral arteries. It is the main cause of death in the western hemisphere, due to cardiovascular syndromes such as myocardial infarction, heart failure, and cerebrovascular accidents. Very substantial economic and human resources have been used on treatments of its complications, including imaging studies, coronary bypass surgery, catheter interventions, pacemakers, and medical treatments. Treating complications, however, are remedial actions. A better alternative is to prevent the development of atherosclerosis, or at least to identify patients who are at risk of acute events and intervene before they occur. The aims of this review are to discuss the predictive value of traditional and emerging risk factors, as well as the role of noninvasive diagnostic methods for coronary atherosclerosis, including exercise stress test, echo stress test, duplex ultrasound, computed tomography, and magnetic resonance. A combination of serum biomarkers and noninvasive approaches is of practical utility for identifying early disease. It is to be expected that future developments will soon perfect our ability to identify the vulnerable patient and allow a more individualized approach. PMID- 16254680 TI - Conservative treatment of pancreatic necrosis with suggestive signs of infection. PMID- 16254681 TI - Use of transobturator suburethral tape for surgical treatment of stress urinary incontinence in a renal transplant patient. PMID- 16254682 TI - Taxonomic significance of leaf anatomy of Aniselytron (Poaceae) as an evidence to support its generic validity against Calamagrostis s. l. AB - A comparative study of leaf anatomy on Aniselytron Merr. and Calamagrostis Adans. s. l. was conducted to review the systematic status of Aniselytron Merr. Calamagrostis s. l. exhibits wide variation in many features, but basic leaf structures of the genus remain constant: absence of a midrib-keel; median and large vascular bundles are central, with double sheaths, accompanied by girders both adaxially and abaxially; prickles have a bulbous base and are not sunken; the abaxial epidermal cells are short and wide and relatively thick-walled. Aniselytron differs from Calamagrostis s. l. in: midrib-keel is present, consisting of a large central bundle with small ones on either side; all vascular bundles are abaxially situated, with abaxial girders only, parenchyma takes the place of the adaxial sclerenchyma; the bases of the prickles are sunken and are not bulbous; the abaxial epidermal cells are tall and thin-walled. These distinct anatomical features, in combination with the differences in spikelet structure and habitat, suggest that Aniselytron should be generically separated from and not merged with Calamagrostis s. l. Due to the adaxial parenchyma in the midrib which has never been found in Pooideae, Aniselytron might have a relationship with some other subfamily. PMID- 16254684 TI - Cytochrome P4501A1 genetic polymorphisms and breast cancer risk in Nigerian women. AB - In this case-control study based on 250 women with breast cancer and 250 age matched controls, we sought to evaluate the role of four polymorphic variants in the CYP1A1 gene in breast cancer susceptibility in Nigerian women. Heterozygosity for the CYP1A1 M1 genotype (CYP1A1 M1 [T/C]) was associated with a 21% reduced risk of breast cancer (OR = 0.79, 95% CI 0.46-1.40) while homozygosity for the genotype (CYP1A1 M1 [C/C]) conferred a non-significant 9% reduced risk of breast cancer. These risk profiles were not significantly altered in subgroup analysis by menopausal status. While heterozygosity for the CYP1A1 M3 genotype (T/C) conferred a non-significant 24% reduced risk of breast cancer (OR = 0.76, 95% CI 0.47-1.22), homozygosity for the variant was associated a non-significant 1.95 fold increased risk of breast cancer (OR = 1.95, 95% CI 0.24-6.01). Subgroup analysis showed a non-significant 11% reduced risk in premenopausal heterozygous carriers (OR = 0.89, 95% CI 0.45-1.44) and a non-significant 6% increased risk of postmenopausal breast cancer for carriers of the CYP1A1 M3 (T/C) genotype. The CYP1A1 M2 (isoleucine to valine) polymorphism in exon 7 and CYP1A1 M4 (threonine to asparagine) variant in codon 461 of the CYP1A1 gene were found to be very rare in our study subjects. This study has shown that while the CYP1A1 M1 polymorphism conferred reduced risk of breast cancer, homozygosity for the CYP1A1 M3 (C/C) was associated with increased risk of breast cancer although these risks did not attain statistical significance. PMID- 16254683 TI - Genome-wide expression analysis of yeast response during exposure to 4 degrees C. AB - Adaptation to temperature fluctuation is essential for the survival of all living organisms. Although extensive research has been done on heat and cold shock responses, there have been no reports on global responses to cold shock below 10 degrees C or near-freezing. We examined the genome-wide expression in Saccharomyces cerevisiae, following exposure to 4 degrees C. Hierarchical cluster analysis showed that the gene expression profile following 4 degrees C exposure from 6 to 48 h was different from that at continuous 4 degrees C culture. Under 4 degrees C exposure, the genes involved in trehalose and glycogen synthesis were induced, suggesting that biosynthesis and accumulation of those reserve carbohydrates might be necessary for cold tolerance and energy preservation. The observed increased expression of phospholipids, mannoproteins, and cold shock proteins (e.g., TIP1) is consistent with membrane maintenance and increased permeability of the cell wall at 4 degrees C. The induction of heat shock proteins and glutathione at 4 degrees C may be required for revitalization of enzyme activity, and for detoxification of active oxygen species, respectively. The genes with these functions may provide the ability of cold tolerance and adaptation to yeast cells. PMID- 16254685 TI - The serine-threonine protein phosphatase PPM1D is frequently activated through amplification in aggressive primary breast tumours. AB - The serine-threonine protein phosphatase PPM1D is likely to play an important role in tumorigenesis. Through inactivation of p38 MAPK, PPM1D acts as a negative feedback regulator of p53 tumour suppressor gene and controls the expression of other cell cycle regulatory proteins, such as CCND1. In addition, recent knock out mouse studies implicated PPM1D in the regulation of p16 expression and the RB tumour suppressor pathway. Here we explored the role of PPM1D aberrations in primary breast cancer. PPM1D copy number analysis showed amplification in 11% (13/117) of the tumours and quantitative real-time RT-PCR revealed a significant correlation (p = 0.0148) between PPM1D amplification and increased expression. PPM1D amplification occurred almost exclusively in tumours with wild-type p53 suggesting that these events are mutually exclusive and further confirming the role of PPM1D as a negative regulator of p53. Interestingly, PPM1D amplification was associated with ERBB2 expression (p = 0.0001) thus implying that PPM1D aberrations occurs in tumours with poor prognosis. We also explored the expression levels of two possible downstream targets of PPM1D. However, immunohistochemical analyses revealed no differences in the staining patterns of CCND1 and p16 proteins in tumours with or without PPM1D aberrations, thus suggesting that previous data from animal model experiments is not directly transferable to primary human tumours. On the other hand, these key cellular proteins are likely to be regulated through a complex fashion in breast cancer and apparently PPM1D represents only one of these mechanisms. Taken together, our findings substantiate an important role for PPM1D in breast cancer. PMID- 16254686 TI - Improvement of breast cancer relapse prediction in high risk intervals using artificial neural networks. AB - The objective of this study is to compare the predictive accuracy of a neural network (NN) model versus the standard Cox proportional hazard model. Data about the 3811 patients included in this study were collected within the 'El Alamo' Project, the largest dataset on breast cancer (BC) in Spain. The best prognostic model generated by the NN contains as covariates age, tumour size, lymph node status, tumour grade and type of treatment. These same variables were considered as having prognostic significance within the Cox model analysis. Nevertheless, the predictions made by the NN were statistically significant more accurate than those from the Cox model (p < 0.0001). Seven different time intervals were also analyzed to find that the NN predictions were much more accurate than those from the Cox model in particular in the early intervals between 1-10 and 11-20 months, and in the later one considered from 61 months to maximum follow-up time (MFT). Interestingly, these intervals contain regions of high relapse risk that have been observed in different studies and that are also present in the analyzed dataset. PMID- 16254687 TI - Complementary and alternative therapeutic approaches in patients with early breast cancer: a systematic review. AB - Complementary and alternative medicine (CAM) is becoming increasingly popular, particularly among patients with breast cancer. We have done a systematic review of studies published between 1995 and February 2005, identified through a comprehensive search. CAM encompasses a wide range of treatment modalities, including dietary and vitamin supplements, mind-body approaches, acupuncture, and herbal medicines. The objectives of CAM treatments are diverse: reduction of therapy-associated toxicity, improvement of cancer-related symptoms, fostering of the immune system and even direct anticancer effects. Clinical trials have generated few or no data on the efficacy of CAM, whether regarding disease recurrence, survival, overall quality of life or safety. Some CAM methods may even have adverse effects or reduce the efficacy of conventional treatment. The primary justification for CAM is based on empirical evidence, case studies, and hypothetical physiological effects. We conclude that available data on CAM modalities in the treatment of early-stage breast cancer does not support their application. PMID- 16254688 TI - VolSurf analysis of pharmacokinetic properties for several antifungal sesquiterpene lactones isolated from Greek Centaurea sp. AB - Sesquiterpene lactones are terpenoid compounds characteristic of the Asteraceae (Compositae) possessing a variety of biological activities, such as cytotoxic, antitumor, antibacterial, and antifungal. The prediction of the pharmacokinetic profile of several antifungal sesquiterpene lactones, isolated from Greek taxa of Centaurea sp., was undertaken in this study using the VolSurf procedure. The molecules were projected on the following pre-calculated ADME models: Caco-2 cell permeability, plasma protein affinity, blood-brain barrier permeation and thermodynamic solubility. The in silico projection revealed a non optimal pharmacokinetic profile for the studied compounds. ADME in silico screening of a semi-synthetic derivatives virtual library has been performed in order to optimize the pharmacokinetic properties. A number of derivatives were proposed as it was predicted to have higher Caco-2 cell permeability, while the pharmacokinetic behaviour regarding BBB penetration, protein binding and solubility was mainly preserved. PMID- 16254689 TI - Dissociation in a polymerization model of homochirality. AB - A fully self-contained model of homochirality is presented that contains the effects of both polymerization and dissociation. The dissociation fragments are assumed to replenish the substrate from which new monomers can grow and undergo new polymerization. The mean length of isotactic polymers is found to grow slowly with the normalized total number of corresponding building blocks. Alternatively, if one assumes that the dissociation fragments themselves can polymerize further, then this corresponds to a strong source of short polymers, and an unrealistically short average length of only 3. By contrast, without dissociation, isotactic polymers becomes infinitely long. PMID- 16254690 TI - Growth of organic microspherules in sugar-ammonia reactions. AB - Reaction of small sugars of less than four carbons with ammonia in water yielded organic microspherules generally less than ten microns in size. The time course of microspherule growth was examined for the D-erythrose-ammonia reaction that yielded microspherules attached to the glass walls of containers. Measurements were made of the elemental composition and infrared spectrum of the microspherule material. These viscose semi-solid microspherules are viewed as possible containers for prebiotic catalytic processes relevant to the origin of life. PMID- 16254691 TI - The last common ancestor: what's in a name? AB - Twenty completely sequenced cellular genomes from the three major domains were analyzed using twice one-way BLAST searches in order to define the set of the most conserved protein-encoding sequences to characterize the gene complement of the last common ancestor of extant life. The resulting set is dominated by different putative ATPases, and by molecules involved in gene expression and RNA metabolism. DEAD-type RNA helicase and enolase genes, which are known to be part of the RNA degradosome, are as conserved as many transcription and translation genes. This suggests the early evolution of a control mechanism for gene expression at the RNA level, providing additional support to the hypothesis that during early cellular evolution RNA molecules played a more prominent role. Conserved sequences related to biosynthetic pathways include those encoding putative phosphoribosyl pyrophosphate synthase and thioredoxin, which participate in nucleotide metabolism. Although the information contained in the available databases corresponds only to a minor portion of biological diversity, the sequences reported here are likely to be part of an essential and highly conserved pool of proteins domains common to all organisms. PMID- 16254692 TI - Habitable zones in the universe. AB - Habitability varies dramatically with location and time in the universe. This was recognized centuries ago, but it was only in the last few decades that astronomers began to systematize the study of habitability. The introduction of the concept of the habitable zone was key to progress in this area. The habitable zone concept was first applied to the space around a star, now called the Circumstellar Habitable Zone. Recently, other, vastly broader, habitable zones have been proposed. We review the historical development of the concept of habitable zones and the present state of the research. We also suggest ways to make progress on each of the habitable zones and to unify them into a single concept encompassing the entire universe. PMID- 16254694 TI - Efferent-mediated fluctuations in vestibular nerve discharge: a novel, positive feedback mechanism of efferent control. AB - We compared the background discharge of vestibular nerve afferents in barbiturate anesthetized and unanesthetized, decerebrate chinchillas. Based on their interspike-interval statistics, units were categorized as regular, intermediate, or irregular. Background discharge rates were higher in irregular units from decerebrates compared to anesthetized preparations; no such difference was observed for regular or intermediate units. Large fluctuations in discharge rate were confined to intermediate and irregular units in decerebrates, but were not seen at all in anesthetized animals. The most prominent examples of fluctuations consisted of oscillations with periods exceeding 500 s and peak-to-peak amplitudes as large as 300 spikes/s. Several observations show that the fluctuations are mediated by the efferent vestibular system (EVS): (1) they are abolished when the vestibular nerve is cut proximal to the recording electrode; (2) their amplitude is correlated with the size of efferent-mediated rotational responses in individual units; and (3) they occur even when vital signs are stable. Previous studies had provided evidence that the EVS involves positive feedback: vestibular nerve afferents and EVS neurons excite one another. To study how oscillations could be produced, we developed a nonlinear model of positive feedback in which afferent feed-forward discharge was nonlinearly related to its inputs from hair cells and the EVS, while these inputs declined (adapted) as discharge was prolonged. Provided that the gain of the efferent feedback loop was sufficiently large, the model showed oscillations similar to those observed experimentally. Although large fluctuations in afferent discharge are unlikely to occur under physiological circumstances, positive feedback may be a normal feature that can amplify the influence of the EVS. PMID- 16254696 TI - Inflammatory demyelination is not central to the pathogenesis of multiple sclerosis. AB - Multiple sclerosis is a disease of the central nervous system that destroys myelin, oligodendrocytes, neurons and axons. Historically considered to be caused by an autoimmune process mainly affecting myelin and oligodendrocytes in the white matter, recent data provide evidence that a generalized, diffuse neurodegenerative process plays an important role in the pathogenesis of MS. There is a high density of axonal transections in active demyelinating lesions, but also persistent low-level axonal damage in inactive plaques and diffuse axonal and neuronal loss throughout the nervous system. Initial axonal injury appears to be closely related to inflammation, but is not restricted to the lesions themselves. Damage may be propagated throughout the nervous system by anterograde Wallerian, retrograde or transynaptic degeneration. Cumulative tissue loss in the grey and white matter, especially of axons, is important and probably the principal determinant of accumulation of irreversible neurological disability and of conversion to a progressive disease course. PMID- 16254697 TI - MRI evidence for multiple sclerosis as a diffuse disease of the central nervous system. AB - The classical view of MS as a chronic inflammatory demyelinating disease leading to the formation of focal central nervous system (CNS) white matter (WM) lesions has been recently challenged by pathological studies and by the extensive application of modern MRI-based techniques. There is now overwhelming evidence supporting the following statements: MS causes widespread tissue damage in the normal-appearing white matter (NAWM) of the brain and spinal cord, whose extent and severity is more strictly associated to the clinical manifestations of the disease than the extent of focal pathology. Discrete, macroscopic lesions are just the tip of the iceberg of MS pathology. Grey matter (GM) damage is a consistent feature of all MS phenotypes, which is progressive from the start of the relapsing-remitting phase of the disease. As is the case for WM, GM damage is also a mixture of focal lesions and diffuse pathology. High-field strength MR scanners are improving our ability to image focal GM lesions and modern MR-based techniques are enabling us to quantify in vivo the extent and severity of GM pathology, which have been shown to correlate only moderately with the amount of WM changes. At least part of GM pathology in MS is not secondary to retrograde degeneration of fibers traversing WM lesions. The neurodegenerative component of the disease is not a late phenomenon and it is not completely driven by inflammatory demyelination. In fact, neurodegeneration occurs very early in the course of MS and the correlation between MRI measures of inflammation and neurodegeneration is weak in all disease phases. The interplay of inflammation and neurodegeneration is a complex and still poorly understood phenomenon. At least part of MS-related neurodegeneration is not directly driven by Wallerian degeneration. Functional cortical changes can be seen in virtually all MS patients and are likely to play a central role in the ability of the MS brain to respond to tissue injury and, hence, limit the functional consequences of structural damage. MS disability is not just the result of tissue destruction but rather a balance between tissue destruction, tissue repair and adaptive cortical reorganization. All of this calls for the concept of MS as a focal, inflammatory demyelinating, WM disease to be reexamined and to start viewing MS as a diffuse CNS disease with an important neurodegenerative component. This is central for identifying novel and effective treatment strategies. PMID- 16254699 TI - The pathology of multiple sclerosis is the result of focal inflammatory demyelination with axonal damage. AB - Multiple sclerosis is a chronic inflammatory demyelinating disease of the central nervous system manifested morphologically by inflammation, demyelination, axonal loss and gliosis. The inflammatory lesions are characterized by massive infiltration by a heterogeneous population of cellular and soluble mediators of the immune system, including T cells, B cells, macrophages and mi croglia, as well as a broad range of cytokines, chemokines, antibodies, complement and other toxic substances. The appearance of such lesions is associated with clinical relapses. Recent detailed immunopathological studies of early, acute lesions revealed profound heterogeneity in the patterns of demyelination and the factors of the immune system involved. During remission, resolution of inflammation is the main factor which leads to clinical improvement of patients. However, the immune system can play a beneficial role at this stage, promoting remyelination perhaps by production of growth factors such as BDNF. In contrast, the progressive irreversible neurological deficit in multiple sclerosis is associated with neurodegenerative processes resulting in axonal and neuronal loss. The mechanisms behind damage to axons in multiple sclerosis lesions are poorly understood. However, the close proximity of areas with prominent axonal loss and areas containing inflammatory infiltrates (e. g., T cells, macrophages) suggest that axonal damage is closely associated with inflammation. Different soluble or cellular mediators of the immune response have been shown to damage axons in experimental systems, and these may be responsible for neurodegeneration in human disease. PMID- 16254698 TI - Changes observed in multiple sclerosis using magnetic resonance imaging reflect a focal pathology distributed along axonal pathways. AB - Multiple sclerosis has long been recognized as a multifocal inflammatory demyelinating disease of the central nervous system. The fact that patients with multiple sclerosis can develop a secondary progressive phase of their disease which is resistant to anti-inflammatory therapies, together with the fact that brain atrophy can develop in patients with a relatively low volume of white matter lesions, has led to suggestions that multiple sclerosis may be a degenerative disease. However, primary degenerative disorders are not usually associated with recurrent episodes of inflammatory demyelination. Support for neurodegeneration in MS being associated with focal lesions comes from topographical mapping of the spatial relationship of axonal injury and tissue loss to lesions using advanced image analysis methods. PMID- 16254700 TI - Purely systemically active anti-inflammatory treatments are adequate to control multiple sclerosis. AB - Collective evidence supports the notion that multiple sclerosis is principally an autoimmune disease. Much of it stems from models of experimental autoimmune encephalomyelitis, generated by inoculation of animals with central nervous system antigens such as MBP, PLP, S100 and MOG or peptides thereof. Different ways of immunization and different animal species and strains mirror different aspects of the neuropathology of multiple sclerosis, such as inflammation, demyelination or axonal damage, and reflect different clinical courses. In all these models, the first immune reactions take place in lymph nodes from which immune cells migrate into the circulation and then to the central nervous system. Adoptive transfer of myelin-reactive T cells from these animals produces pathology and disease in the central nervous system of naive healthy recipients. In the human disease, autoreactive T and B cells specific for a variety of central antigens are present in the immune repertoire. These cells appear to be activated in the periphery through a number of mechanisms which causes them to home to the central nervous system. Contact with the local immune circuitry of the brain stimulates clonal expansion of autoreactive T cells, initiating a cascade of immuno-inflammatory events in situ. Numerous ways of disrupting this complex sequence of events, either by non-specific immunosuppression or by targeting specific checkpoints, abrogate or ameliorate disease in animal models. All approved disease-modifying drugs have an impact on components of the systemic immune compartment. All have been shown to reduce the number of gadolinium enhancing T1 lesions observed with magnetic resonance imaging, an index of acute inflammatory invasion of the central nervous system. PMID- 16254701 TI - Modulating processes within the central nervous system is central to therapeutic control of multiple sclerosis. AB - Historically considered to be an autoimmune demyelinating disease, multiple sclerosis is now recognized to be characterized by significant axonal and neuronal pathology. Addressing this neurodegenerative component of the disease is an important treatment objective, since axonal injury is believed to underlie the accumulation of disability and disease progression. The precise relationship between the inflammatory and neurodegenerative components in multiple sclerosis remains poorly elucidated, although neurodegeneration appears to be at least partially independent from neuroinflammation. The mechanisms underlying axonal injury appear complex and are likely to be multifactorial. Specific treatment strategies need to be developed that act within the central nervous system to prevent neurodegeneration and need to be provided from the earliest stages of disease. It is likely that immunomodulatory treatments acting purely in the periphery will provide only indirect and not direct neuroprotection. A promising approach is to enhance neuroprotective autoimmunity inside the brain, believed to be mediated, at least in part, by the release of neurotrophic factors within the nervous system from infiltrating immune cells. Such a beneficial process would be inhibited by a non-selective immunosuppressive strategy. In summary, treatments of multiple sclerosis should take into account the heterogeneous pathophysiology of the disease. The pathogenic process in the central nervous system itself should be the major focus in multiple sclerosis therapy in order to protect against demyelination and axonal loss and to promote remyelination and regeneration directly in the target tissue, independently of peripheral immune status. In conclusion, selective treatment strategies aimed at preventing axonal injury within the central nervous system are required to complement existing, peripherally acting treatments targeting the immune system. PMID- 16254702 TI - Multiple sclerosis trial designs for the 21st century: building on recent lessons. AB - Starting with the first positive pilot study of glatiramer acetate, trial design in multiple sclerosis has advanced considerably over the past two decades, successively building and improving on previous successes in the implementation and analysis of new clinical trials. Most of these trials have been successful and this has led to the regulatory approval and commercial availability of six agents for the treatment of multiple sclerosis. During this period, outcome measures have been validated to determine the efficacy and safety of such agents, notably those useful in reducing the inflammatory aspects of disease. These include measurements of relapse reduction (annualized relapse rate, time to first relapse, proportion of subjects relapse free), disability (change in EDSS score, change in MSFC score) and MRI metrics (measurements of gadolinium-enhancing lesions, T1 and T2 lesion load). Recent trial design has shown that one can answer some clinical questions after one year on study and that these results may be predictive of more robust two-year trial data. The other important recent lesson involves emergence of rare complications of immunomodulatory therapy, namely progressive multifocal leucoencephalopathy with natalizumab that blocks the access of immune cells to the nervous system. In addition to the increased need for enhanced safety assessment, this issue will have an impact both on the study of combination therapies and on the use of combinations in clinical practice. PMID- 16254703 TI - Effects of insulin-like growth factor-1 (IGF-1) and amifostine in spinal cord reirradiation. AB - PURPOSE: To test whether insulin-like growth factor-1 (IGF-1) and amifostine modulate the reirradiation tolerance of rat cervical spinal cord. Initial experiments by the authors' group suggested that administration of each agent alone significantly increased the median latent time to radiation myelopathy (RM) in previously unirradiated animals but did not change the dose-response relationship. Because of different modes of action, a follow-up study was undertaken to test the combined treatment. MATERIAL AND METHODS: The cervical spinal cord of 51 adult Fisher F-344 rats received a single fraction of 16 Gy, which corresponds to approximately 75% of the median paresis dose (ED(50)), followed 5 months later by a second radiation dose, which ranged from 17 to 21 Gy. The study animals received a single intrathecal injection of 0.3 mg amifostine into the cisterna magna 30-60 min before reirradiation plus three subcutaneous doses of IGF-1 (700 microg) starting from 24 h before to 24 h after reirradiation. Control animals received saline injections via the same routes. Animals were followed until RM developed or until 12 months from reirradiation. Histopathologic examinations were performed post mortem. RESULTS: No animals showed any neurologic abnormalities before reirradiation. RM occurred in controls versus treated rats after a mean latency of 218 versus 314 days (19 Gy; p = 0.11) and 104 versus 186 days (21 Gy; p = 0.002) from second dose (Figure 1). ED(50) was 18.2 versus 19.4 Gy (p = 0.15; Figure 2). Treatment with IGF-1 and amifostine did not significantly influence the rates of tumor induction or intercurrent death. CONCLUSION: IGF-1 and amifostine significantly reduced RM rates after 21 Gy. The shift of the dose-response curve suggests an increase of the ED(50) for single-dose treatment by approximately 7%. Thus, brief therapeutic intervention might slightly decrease radiation-induced neurotoxicity in a retreatment situation. PMID- 16254704 TI - Feasibility and early results of interstitial intensity-modulated HDR/PDR brachytherapy (IMBT) with/without complementary external-beam radiotherapy and extended surgery in recurrent pelvic colorectal cancer. AB - BACKGROUND: A new multimodality treatment concept consisting of extended resection and postoperative fractionated intensity-modulated interstitial brachytherapy (IMBT) was introduced for pelvic recurrence of colorectal carcinoma. PATIENTS AND METHODS: 46 patients received extended resection and single plastic tubes were sutured directly onto the tumor bed. IMBT was started within 2 weeks postoperatively with a median dose of 24.5 Gy (5-35 Gy). Patients were treated either with high-dose-rate brachytherapy (HDR; n = 23) or with pulsed-dose-rate brachytherapy (PDR; n = 23). 25 patients received complementary 45-Gy external-beam irradiation (EBRT) to the pelvic region after explanting the plastic tubes. RESULTS: Median follow-up was 20.6 months (7-107 months) and mean patient survival 25.7 +/- 25.8 months (median 17, range 1-107 months). After 5 years overall survival, disease-free survival and local control rate were 23%, 20% and 33%, significantly influenced by the resectional state. There was a trend in favor of PDR compared to HDR, which reached statistical significance in patients who had not received additional EBRT. CONCLUSION: The combination of extended surgery and postoperative interstitial IMBT is feasible and offers effective interdisciplinary treatment of recurrent colorectal cancer. In this small and inhomogeneous cohort of patients PDR seems to be more effective than HDR, particularly when application of complementary EBRT is not possible. None of the patients who required resection of distant metastasis survived > 2 years in this study. PMID- 16254705 TI - Renal dysfunction after total-body irradiation. Significance of selective renal shielding blocks. AB - PURPOSE: A retrospective analysis was conducted on the outcome of total-body irradiation (TBI) followed by bone marrow transplantation (BMT) on leukemia patients. Also studied was the risk of renal dysfunction after TBI/BMT with or without the use of selective renal shielding blocks. PATIENTS AND METHODS: The cases of 109 leukemia patients who received TBI as a component of the conditioning regimen for their BMT were reviewed. They received 12 Gy of TBI in six fractions over 3 consecutive days. Doses to eyes and lungs were reduced to 7 Gy and 8 Gy, respectively, by customized organ shielding blocks. After March 1999, renal shielding blocks were used to constrain the renal dose to 10 Gy. The patients were followed for a median period of 16.6 months (range: 0.3-180.1 months). RESULTS: The 2-year and 5-year overall survival rates were 55.4% and 43.2%, respectively. Renal dysfunction-free rates were different between those with and without renal shielding blocks: 100% and 78.5%, respectively, at 2 years. Overall survivals were not significantly different among these patients: 60.4% and 52.9%, respectively, at 2 years in patients with and without renal shielding blocks (p = 0.53). CONCLUSION: The use of selective renal shielding blocks provided evidence for reducing radiation-induced renal toxicities without decreasing the overall survival rate. PMID- 16254706 TI - The impact of varying volumes in rectal balloons on rectal dose sparing in conformal radiation therapy of prostate cancer. A prospective three-dimensional analysis. AB - BACKGROUND AND PURPOSE: The use of a rectal balloon leads to a protection of the posterior rectal wall in irradiation of prostate cancer. The purpose of this analysis was to quantitatively assess the optimal volume in rectal balloons concerning rectal dose sparing in different clinical target volumes (CTVs) in prostate cancer irradiation. PATIENTS AND METHODS: 14 patients with localized prostate cancer undergoing external beam radiotherapy were investigated. The prostate, the entire and the proximal seminal vesicles were delineated as CTV. Treatment plans without a rectal balloon and with a rectal balloon inflated with 40 ml (six patients) or 60 ml air (eight patients) were generated for each CTV and compared concerning rectal dose volume histograms. RESULTS: The use of a rectal balloon filled with 40 ml air led to no significant advantage in radiation exposure of the rectal wall in all CTVs. The use of a rectal balloon filled with 60 ml air resulted in a significant decrease of the exposed rectal wall volume in all CTVs with a reduced estimated risk for chronic toxicity in case of inclusion of the proximal or entire seminal vesicles into the CTV. CONCLUSION: The use of a rectal balloon filled with 60 ml air led to a significantly decreased proportion of the irradiated rectal wall for all CTVs. This volume filled in rectal balloons is therefore recommended for use. In case of irradiation of the prostate without the seminal vesicles, the use of a rectal balloon should be considered carefully concerning the patients' imaginable discomfort using a rectal balloon and a questionable advantage concerning the estimated risk for chronic toxicity. PMID- 16254707 TI - Biologically effective doses of postoperative radiotherapy in the prevention of keloids. Dose-effect relationship. AB - PURPOSE: To review the recurrence rates of keloids after surgical excision followed by radiotherapy, and to answer the question whether after normalization of the dose, a dose-effect relationship could be derived. MATERIAL AND METHODS: A literature search was performed to identify studies dealing with the efficacy of various irradiation regimens for the prevention of keloids after surgery. Biologically effective doses (BEDs) of the various irradiation regimens were calculated using the linear-quadratic concept. A distinction between recurrence rates of keloids in the face and neck region and those in other parts of the body was made. RESULTS: 31 reports were identified with PubMed with the search terms keloids, surgery, radiation therapy, radiotherapy. 13 reports were excluded, because no link could be found between recurrence rate and dose, or if less than ten patients per dose group. The recurrence rate for surgery only was 50-80%. For BED values > 10 Gy the recurrence rate decreased as a function of BED. For BED values > 30 Gy the recurrence rate was < 10%. For a given dose, the recurrence rates of keloids in the sites with high stretch tension were not significantly higher than in sites without stretch tension. CONCLUSION: The results of this study indicate that for effectively treating keloids postoperatively, a relatively high dose must be applied in a short overall treatment time. The optimal treatment probably is an irradiation scheme resulting in a BED value of at least 30 Gy. A BED value of 30 Gy can be obtained with, for instance, a single acute dose of 13 Gy, two fractions of 8 Gy or three fractions of 6 Gy, or a single dose of 27 Gy at low dose rate. The radiation treatment should be administered within 2 days after surgery. PMID- 16254708 TI - Results of postoperative (90)Sr radiotherapy of keloids in view of patients' subjective assessment. AB - BACKGROUND AND PURPOSE: As treatment of keloids is mainly a cosmetic indication, the authors investigated, beyond the recurrence rate, the patients' satisfaction with the result and its correlation with objective medical findings. PATIENTS AND METHODS: 83 keloids of 66 patients had been irradiated after excision by a uniform protocol with 4 x 5 Gy (strontium- 90 [(90)Sr] surface applicator). A questionnaire was developed and sent out in which, above all, the satisfaction with the therapeutic and cosmetic outcome was obtained. These results were correlated with objective parameters and medical findings which were ascertained during an extra follow-up examination. RESULTS: Among 18 of the 41 patients (44%), who had answered the questionnaire, 19 of the 53 keloids treated (36%) had relapsed. 61% of the patients were extremely or mainly satisfied with the therapeutic outcome, 51% extremely or mainly satisfied with the cosmetic outcome. The relief from former keloid-caused symptoms (therapeutic outcome: p = 0.0005; cosmetic outcome: p = 0.0011), the ear as keloid localization (p = 0.0008 and p = 0.0197), and male gender (therapeutic outcome: p = 0.0423) were significantly associated with higher satisfaction. The recurrence rate as well as the extent of radiation side effects had no significant influence on patients' assessment. CONCLUSION: Cosmetic aspects like the dermal side effects and the patients' satisfaction should be taken into account when evaluating the results of radiotherapy in keloids. PMID- 16254709 TI - No effect of the hemoglobin solution HBOC-201 on the response of the rat R1H tumor to fractionated irradiation. AB - BACKGROUND AND PURPOSE: Tumor hypoxia is regarded as one important underlying feature of radioresistance. The authors report on an experimental approach to improve tumor response to radiation by combining fractionated irradiation with HBOC-201, an ultrapurified polymerized hemoglobin solution, which is currently used in clinical phase II/III trials as alternative oxygen carrier and proved to be highly effective in tissue oxygenation (tpO(2)). MATERIAL AND METHODS: Subcutaneously growing rhabdomyosarcoma R1H tumors of the rat were treated with either 40 Gy (2 Gy/fraction, 20 fractions in 2 weeks, ambient) followed by graded top-up doses (clamped) alone, or in combination with HBOC-201, or with HBOC-201 plus carbogen (95% O(2) + 5% CO(2)). Local tumor control (TCD50%) and growth delay were used as endpoints. In addition, the effect of HBOC-201 alone or in combination with carbogen on the tpO(2) of tumor and muscle was determined using a flexible stationary probe (Licox, GMS). RESULTS: TCD50% values of 119 Gy (95% confidence interval 103;135), 111 Gy (84;138), and 102 Gy (83;120) were determined for tumors irradiated alone, in combination with HBOC-201, and with HBOC-201 plus carbogen, respectively. Although the dose-response curves showed a slight shift to lower doses when HBOC-201 or HBOC-201 plus carbogen was added, the differences in TCD50% were not statistically significant. No effect was seen on the growth delay of recurrent tumors. HBOC-201 alone did not effect tumor or muscle tpO(2). In combination with carbogen the mean tpO(2) of muscle raised from 23.9 mmHg to 59.3 mmHg (p < 0.05), but this effect was less pronounced than the increase in tpO(2) by carbogen alone. CONCLUSION: Low-dose application of HBOC 201 does not improve the response of the rhabdomyosarcoma R1H of the rat to fractionated irradiation. PMID- 16254710 TI - [Commentary of F. Kahmann, T. Henkel, P. Wust, D. Bohmer zum Beitragon the article by C. Hoinkis et al. "Evaluation of dose volume histograms after prostate seed implantation. 4 year experience" in: Strahlenther Onkol 2004;180:550-6 (No. 9)]. PMID- 16254712 TI - Comment by E. Weiss and C. F. Hess on G. M. Villeirs et al. "Interobserver delineation variation using CT versus combined CT + MRI in intensity-modulated radiotherapy for prostate cancer" in: Strahlenther Onkol 2005;181:424-30 (No. 7). PMID- 16254714 TI - Osteochondritis (Calve's disease) of a vertebral body: a rare form of vertebra plana (T. Shisha et al.). PMID- 16254715 TI - Does it take two to tango? PMID- 16254718 TI - Effects of chronic exercise conditioning on thermal responses to lipopolysaccharide and turpentine abscess in female rats. AB - Chronic exercise conditioning has been shown to alter basal thermoregulatory processes as well as the response to inflammatory agents. Two such agents, lipopolysaccharide (LPS) and turpentine (TPT) are inducers of fever in rats. LPS, given intraperitoneally (i.p.), involves a systemic inflammatory response whereas TPT given intramuscularly (i.m.) elicits a localized inflammation. We assessed if chronic exercise training in the rat would alter the thermoregulatory response to LPS and TPT. Core temperature (T (c)) and motor activity were monitored by radiotelemetry. Female Sprague Dawley rats were divided into two groups (trained and sedentary) and housed at an ambient temperature of 22 degrees C. Animals voluntarily trained on running wheels for 8 weeks. In the first study, trained and sedentary female rats were injected i.p. with LPS (50 microg/kg) or an equal volume of 0.9% normal saline. In another study, trained and sedentary female rats were injected i.m. with TPT (10 microl)/rat or an equal volume of 0.9% normal saline. The time course of the LPS fever was very short compared to TPT. TPT injected animals displayed a smaller but more prolonged fever compared to LPS; however, training accentuated the febrile response to LPS (DeltaT (c)=0.6 degrees C in sedentary and 1.2 degrees C in trained). Training had a slight suppression on TPT-induced fever during the daytime but had no effect on motor activity or nighttime T (c). In contrast, exercise training led to a marked increase in the pyrogenic effects of LPS. We conclude that the effect of exercise training and source of infection (i.e., systemic versus localized in muscle) on fever is directly linked to type of pyrogenic agent. PMID- 16254717 TI - The role of toll-like receptor 4 in airway inflammation induced by diesel exhaust particles. AB - Although several studies have demonstrated that airway exposure to diesel exhaust particles (DEP) induces lung inflammation, the signaling pathways involved in the pathogenesis remain unclear. Toll-like receptors (TLRs) are generally accepted to be pathogen recognition receptors in mammalians. In the present study, we investigated the role of TLR-4 in DEP-induced lung inflammation and cytokine expression in the lung in TLR-4 point mutant (C3H/HeJ) mice and corresponding control (C3H/HeN) mice. Both the types of mice were randomized into four experimental groups that received vehicle or DEP (12 mg/kg body weight) by intratracheal instillation (n = 8-10 in each group). Cellular profile of bronchoalveolar lavage (BAL) fluid, expressions of cytokines and chemokines in the lung, and circulatory fibrinogen levels were evaluated 24 h after the instillation.DEP challenge revealed a significant increase in the numbers of total cells and neutrophils in the BAL fluid as compared to vehicle challenge, however, the numbers were less in C3H/HeJ mice than in C3H/HeN mice. DEP exposure significantly induced the lung expression of interleukin (IL)-1beta, keratinocyte chemoattractant (KC), and macrophage inflammatory protein (MIP)-1alpha when compared to vehicle challenge in both genotypes of mice. In the presence of DEP, the level of MIP-1alpha was significantly lower in C3H/HeJ mice than in C3H/HeN mice, however, the levels of IL-1beta, KC, and fibrinogen showed opposite findings. These results suggest that TLR-4 is one of recognition receptors against DEP in the airways. PMID- 16254716 TI - Charite total disc replacement--clinical and radiographical results after an average follow-up of 17 years. AB - A retrospective clinical-radiological study to evaluate the long-term outcome after artificial disc replacement was performed. The objective is to investigate long-term results after implantation of a modular type artificial disc prosthesis in patients with degenerative disc disease (DDD). Total disc replacement (TDR) is a surgical procedure intended to save segmental spinal function, and thus replace spondylodesis. Short-term results are promising, whereas long-term results are scarce. The Charite TDR is the oldest existing implant, therefore, the longest possible follow-up is presented here. Seventy-one patients were treated with 84 Charite TDRs types I-III. Indication for TDR was moderate to severe DDD. Fifty three patients (63 TDRs) were available for long-term follow-up of 17 years. Evaluation included Oswestry disability index, visual analog scale, overall outcome score, plain and extension/flexion radiographs. Implantation of Charite TDR resulted in a 60% rate of spontaneous ankylosis after 17 years. No significant difference between the three types of prostheses was found concerning clinical outcome. Reoperation was necessary in 11% of patients. Although no adjacent segment degeneration was observed in the functional implants (17%), these patients were significantly less satisfied than those with spontaneous ankylosis. TDR, nowadays, is an approved procedure. Proof that long-term results of TDR implantation in DDD are at least as good as fusion results is still missing. PMID- 16254719 TI - Interfacing liquid chromatography with atmospheric pressure MALDI-MS. AB - Two different strategies for coupling liquid chromatography with atmospheric pressure matrix assisted laser desorption/ionization (AP MALDI) are presented. The first method is flow-injection liquid AP UV-MALDI. Compared with previous similar research, the detection limit was improved 10 times to 8.3 fmol using a solution of 50 nM peptide with 25 mM alpha-cyano-4-hydroxycinnamic acid. The applicability of this method to measure oligosaccharides, actinomycin antibiotics, antibiotics, phosphopeptides, and proteins is demonstrated. The upper mass limit achieved with the current instrumentation is 6,500 Da (doubly charged cytochrome c). The feasibility of a second strategy based on single droplet IR AP MALDI is demonstrated here. Aqueous peptide solutions were successfully measured by this method. PMID- 16254721 TI - Direct competitive ELISA based on a monoclonal antibody for detection of aflatoxin B1. Stabilization of ELISA kit components and application to grain samples. AB - A direct competitive enzyme-linked immunosorbent assay (ELISA) based on a monoclonal antibody has been developed and optimized for detection of aflatoxin B1 (AFB1), and an ELISA kit has been designed. This immunoassay was highly specific, sensitive, rapid, simple, and suitable for aflatoxin monitoring. AFB1 concentrations determinable by ELISA ranged from 0.1 to 10 microg L(-1). The IC50 value was 0.62 microg L(-1). Recovery from spiked rice samples averaged between 94 and 113%. The effect of different reagents on the stability of HRP-AFB1 conjugate solution was studied. The performance of a stabilized enzyme tracer in ELISA was determined and compared with that of a freshly prepared control solution of HRP-AFB(1) conjugate. The results showed that stabilizing media containing 0.02% BSA, 0.1% Kathon CG, and 0.05 mol L(-1) calcium chloride in 0.05 mol L(-1) Tris-HCl buffer (pH 7.2) maintained the activity of HRP-AFB1 at a dilution of 1:1000 for a period of at least 12 months at room temperature whereas the reference conjugate solution without the additives lost its activity within a few days. Several additives were tested for their stabilizing effect on a monoclonal antibody (MAb) immobilized on the surface of polystyrene microtitre plates. It was shown that immobilized MAb, treated with post-coating solutions containing PVA, BSA, and combinations of these substances with trehalose, retained its activity for at least 4 months at 4 degrees C, whereas the untreated MAb-coated plate lost its activity within 2 days. PMID- 16254722 TI - [Postoperative pain relief is an important factor for the patients' selection of a clinic. Results of an anonymous survey]. AB - AIM: The aim of the survey was to elucidate the significance of postoperative pain therapy for the patient and its influence on the choice of hospital. METHODS: This prospective, anonymous survey of consecutive patients in a general surgical clinic was performed by an independent study nurse. RESULTS: A total of 161 patients were included; 90% of the surgical patients considered "good pain therapy" as a highly important factor, and three of four patients would admit themselves more likely to a clinic well known for "good pain therapy." If the patients could choose their hospital, factors such as "medical care by the physicians" are most important (rank 1; rank 1 most important, rank 10 most unimportant) followed by quality of "nursing care" (mean rank 2.6) and "good pain therapy" (mean rank 3.6). Older patients (> or = 60 years) preferred a hospital with known "good pain therapy" more often for surgical therapy than younger patients (< 60 years). CONCLUSION: It can be concluded that postoperative pain relief is an important factor for the patients' selection of a clinic and is influenced by an age of > or = 60 years. PMID- 16254723 TI - [Referred pain from amputation stump trigger points into the phantom limb]. AB - OBJECTIVES: The aim of the study was to examine whether/how myofascial stump trigger points (TPs) after lower leg amputations are able to produce stump pain (SP), phantom pain (PP), and sensations (PS) in the phantom limb. METHODS: Palpation of the 5 most striking stump TPs of 30 leg amputees (12 transfemoral, 18 transtibial) was documented in a standardized manner. Patients were asked to localize SP, PP, and PS. RESULTS: Of 150 TPs, 14 produced involuntary stump movements and 10 stump fasciculations. Dorsal PP after ventral TP palpation occurred as well as PP in the toes from TPs near the hip. Of 30 patients, 20 reported PS and 8 PP; 60 of 150 TPs produced PS and 17 PP. Phantom phenomena were localized in 62.8% in the toes (1st toe 19.8%, toes 2-5 about 10% each), 17.9% midfoot, and the rest were more proximal. TPs were localized more in the lateral/dorsal stump than medial/ventral. About 70% of the TPs were found between 3 and 7 cm from the stump end, those with toe projections more distal than those with tibial projections. CONCLUSIONS: Myofascial TPs in amputation stumps are common and able to produce sensations and pain in the phantom limb. Most reported experiences were localized in the toes, as phantom pain usually is. There seems to be a "stump representation" and it seems possible that "referred TP pain" and "phantom pain" may develop from similar origin. PMID- 16254724 TI - Developmental and gene expression analyses of a cotton naked seed mutant. AB - Cotton fiber development is a fundamental biological phenomenon, yet the molecular basis of fiber cell initiation is poorly understood. We examined molecular and cellular events of fiber cell development in the naked seed mutant (N1N1) and its isogenic line of cotton (Gossypium hirsutum L. cv. Texas Marker-1, TM-1). The dominant mutation not only delayed the process of fiber cell formation and elongation but also reduced the total number of fiber cells, resulting in sparsely distributed short fibers. Gene expression changes in TM-1 and N1N1 mutant lines among four tissues were analyzed using spotted cotton oligo-gene microarrays. Using the Arabidopsis genes, we selected and designed approximately 1,334 70-mer oligos from a subset of cotton fiber ESTs. Statistical analysis of the microarray data indicates that the number of significantly differentially expressed genes was 856 in the leaves compared to the ovules (3 days post anthesis, DPA), 632 in the petals relative to the ovules (3 DPA), and 91 in the ovules at 0 DPA compared to 3 DPA, all in TM-1. Moreover, 117 and 30 genes were expressed significantly different in the ovules at three and 0 DPA, respectively, between TM-1 and N1N1. Quantitative RT-PCR analysis of 23 fiber-associated genes in seven tissues including ovules, fiber-bearing ovules, fibers, and non-fiber tissues in TM-1 and N1N1 indicates a mode of temporal regulation of the genes involved in transcriptional and translational regulation, signal transduction, and cell differentiation during early stages of fiber development. Suppression of the fiber-associated genes in the mutant may suggest that the N1N1 mutation disrupts temporal regulation of gene expression, leading to a defective process of fiber cell elongation and development. PMID- 16254725 TI - Morphogenesis and molecular basis on naked seed rice, a novel homeotic mutation of OsMADS1 regulating transcript level of AP3 homologue in rice. AB - The floral organs are formed from floral meristem with a regular initiation pattern in angiosperm species. Flowers of naked seed rice (nsr) were characterized by the overdeveloped lemma and palea, the transformation of lodicules to palea-/lemma-like organs, the decreased number of stamens and occasionally extra pistils. Some nsr spikelets contained additional floral organs of four whorls and/or abnormal internal florets. The floral primordium of nsr spikelet is differentiated under an irregular pattern and an incomplete determination. And molecular analysis indicated that nsr was a novel homeotic mutation in OsMADS1, suggesting that OsMADS1 played a distinct role in regulating the differentiation pattern of floral primordium and in conferring the determination of flower meristem. The gain-of-function of OsMADS1 transgenic lines presented the transformation of outer glumes to lemma-/palea-like organs and no changes in length of lemma and palea, but loss-of-function of OsMADS1 transgenic lines displayed the overdeveloped lemma and palea. Both findings revealed that OsMADS1 played a role in specifying lemma and palea and acted as a repressor of overdevelopment of lemma and palea. Moreover, it was indicated that OsMADS1 upregulated the transcript level of AP3 homologue OsMADS16, using real time PCR analysis on gain- and loss-of-function of OsMADS1 transgenic lines. PMID- 16254726 TI - Associations of psychosocial working conditions with self-rated general health and mental health among municipal employees. AB - OBJECTIVES: To examine associations of job demands and job control, procedural and relational organizational fairness, and physical work load with self-rated general health and mental health. In addition, the effect of occupational class on these associations is examined. METHODS: The data were derived from the Helsinki Health Study baseline surveys in 2001-2002. Respondents to cross sectional postal surveys were middle-aged employees of the City of Helsinki (n=5.829, response rate 67%). Associations of job demands and job control, organizational fairness and physical work load with less than good self-rated health and poor GHQ-12 mental health were examined. RESULTS: Those with the poorest working conditions two to three times more, often reported poor general and mental health than those with the best working conditions. Adjustment for occupational class weakened the associations of low job control and physical work load with general health by one fifth, but even more strengthened that of high job demands. Adjustment for occupational class clearly strengthened the associations of job control and physical work load with mental health in men. Mutual adjustment for all working conditions notably weakened their associations with both health measures, except those of job control in men. All working conditions except relational organizational fairness remained independently associated with general and mental health. CONCLUSIONS: All studied working conditions were strongly associated with both general and mental health but the associations weakened after mutual adjustments. Of the two organizational fairness measures, procedural fairness remained independently associated with both health outcomes. Adjustment for occupational class had essentially different effects on the associations of different working conditions and different health outcomes. PMID- 16254728 TI - Analysis of avian bone response to mechanical loading-Part one: Distribution of bone fluid shear stress induced by bending and axial loading. AB - Mechanical loading-induced signals are hypothesized to be transmitted and integrated by a bone-connected cellular network (CCN) before reaching the bone surfaces where adaptation occurs. Our objective is to establish a computational model to explore how bone cells transmit the signals through intercellular communication. In this first part of the study the bone fluid shear stress acting on every bone cell in a CCN is acquired as the excitation signal for the computational model. Bending and axial loading-induced fluid shear stress is computed in transverse sections of avian long bones for two adaptation experiments (Gross et al. in J Bone Miner Res 12:982-988, 1997 and Judex et al. in J Bone Miner Res 12:1737-1745, 1997). The computed fluid shear stress is found to be correlated with the radial strain gradient but not with bone formation. These results suggest that the radial strain gradient is the driving force for bone fluid flow in the radially distributed lacunar-canalicular system and that bone formation is not linearly related to the loading-induced local stimulus. PMID- 16254727 TI - Efficacy of 1-day prophylaxis medication with fluoroquinolone for prostate biopsy. AB - A prospective, randomized trial in which 236 patients received oral levofloxacin, either at 600 mg/day for 1 day (n = 124) or 300 mg/day for 3 days (n = 112). Urinalysis, plasma white blood cell count (WBC) (per mm3), and C reactive protein (CRP) (mg/dl) levels were checked before prostate biopsy (PBX), on the day after PBX, and on the seventh day after PBX. Two patients (1.61%) who received 600 mg for 1 day and 2 patients (1.79%) who received 300 mg for 3 days had febrile infectious complications. There was no statistically significant difference between levofloxacin at 600 mg for 1 day and levofloxacin at 300 mg for 3 days regarding the elevation of WBC and CRP. We can perform PBX safely with levofloxacin at 600 mg for 1 day as prophylaxis and recommend this method from the point of view of the decrease of antibiotic-resistant strains. PMID- 16254729 TI - An unusual case of homicidal near drowning followed by electrocution. AB - A young woman was found dead in a bath-tub with her facial orifices under water and current burn marks at the right big toe and at the neck near the hairline. Initial police investigations assumed an accidental electric shock due to a radio falling into the water. The forensic autopsy was followed by extensive histology and immunohistochemistry of the internal organs and skin marks. Intensive vital reactions of a near-drowning were detected that suggested a survival period of at least 20 min. Furthermore, an energy dispersive x-ray spectroscopy analysis on the current burn marks and on possible current applicators was performed, as well as reconstruction experiments with a cadaver of a pig that had been slaughtered shortly before. The results of all investigations revealed a two-step procedure with a primary near-drowning and a final current application with consecutive cardiac arrest. PMID- 16254730 TI - Effect of stimulants on 24-h ambulatory blood pressure in children with ADHD: a double-blind, randomized, cross-over trial. AB - Millions of children with attention deficit hyperactivity disorder (ADHD) are treated with stimulant medications. To evaluate cardiovascular risk, 24-h ambulatory blood pressure monitoring (ABPM) was performed on and off medication. Thirteen subjects underwent APBM both on stimulant therapy and placebo using a placebo-controlled, double-blind, randomized, cross-over design. After a 3-day run-in followed by a 24-h monitoring period, subjects crossed over to the alternate therapy for repeated ABPM. Subjects demonstrated elevations in most hemodynamic parameters derived from ABPM during the active treatment period. Total diastolic blood pressure (69.7 mmHg vs 65.8 mmHg, p =0.02) and waking diastolic blood pressure (75.5 mmHg vs 72.3 mmHg, p =0.03) were significantly higher during active treatment. Total heart rate was also significantly higher during active treatment (85.5 beats/min vs 79.9 beats/min, p =0.004). The rate pressure product (the product of systolic blood pressure x heart rate), an index of myocardial oxygen demand, was higher during active treatment (9,958 vs 9,076, p =0.008). This study provides evidence for a possible negative cardiovascular effect of stimulant medications in children with ADHD. This potential cardiovascular risk should be balanced against the beneficial behavioral effects of this class of medication. PMID- 16254731 TI - Timed-urine collections for renal clearance studies. AB - The purpose of this study was to describe the reproducibility of timed-urine collections for renal clearance studies and the effect variations in urine collection has on measurement of glomerular filtration rate (GFR). Data from 222 cimetidine clearance studies (GFR-Cim) were obtained from 32 pediatric renal patients over a period of 8 years. There were three to 18 studies per child aged 4.8 years to 21 years at the time of a study. The urinary creatinine excretion rate is measured during supervised urine collection periods. The creatinine excretion rates in each child were compared to obtain data on the reproducibility of the urine collections. The coefficient of variation (CV) of the creatinine excretion rate is approximately 10% in both children and adults. The variation in GFR to be expected during repeated renal clearance studies in subjects with stable GFR, using voided urine collections, was similar in children and adults, with a CV of 12% to 14%. PMID- 16254732 TI - Prediction in the timing of pursuit eye movement initiation revealed by cross axis vestibular-pursuit training in monkeys. AB - The smooth-pursuit system interacts with the vestibular system to maintain the image of a moving target on the fovea. Efficient tracking performance requires information about the velocity and the initiation of target motion. Previous studies in monkeys have shown that training with orthogonal pursuit and whole body rotation results in adapted eye movement direction during chair rotation. In addition, the latency of the pursuit shortens and initial eye velocity increases in a task-dependent manner. To examine whether these adapted eye movements are predictive pursuit, we studied whether our monkeys could predict the timing of smooth eye movement initiation during chair rotation. Two young Japanese monkeys were rotated horizontally in a trapezoidal waveform (20 degrees/s, +/-10 degrees) with random inter-trial intervals. A laser spot was moved vertically with the same trajectory at a constant delay ranging from 100 to 700 ms after the onset of the chair motion. The monkeys were required to pursue the spot. After this training, the latencies of pursuit eye movements following the onset of chair motion were examined in the presence of the target motion. The target was also briefly (for 500-700 ms) extinguished at 80 ms after the onset of chair rotation. Pursuit eye movements after training were initiated before the onset of target motion and the latencies were proportional to the delays used for training. The latencies and response magnitudes of pursuit with or without target blanking were similar. The auditory-pursuit training did not induce an initial pursuit response similar to that induced by vestibular-pursuit training. These results indicate that smooth eye movements during the chair rotation after the vestibular-pursuit training included a predictive pursuit component. The monkeys' estimate of the delays revealed by the latencies of pursuit was shorter by 22-36% than the actual delays. PMID- 16254733 TI - Inhibition of cytokinesis and akt phosphorylation by chaetoglobosin K in ras transformed epithelial cells. AB - PURPOSE: Chaetoglobosin K (ChK), a bioactive natural product previously shown to have anti-tumor promoting activity in glial cells and growth inhibitory effects in ras-transformed fibroblasts, inhibited anchorage-dependent and anchorage independent growth in ras-transformed liver epithelial cells. The purpose of this study was to identify cellular targets of ChK that mediate its anti-tumor effects. METHODS: Anchorage-independent cell growth assays, using soft agar coated dishes, and anchorage-dependent growth assays were performed on transformed WB- ras1 cells. Phase/contrast and fluorescent microscopy were used to visualize cell morphological changes and DAPI-stained nuclei. Analyses of p21 Ras membrane versus cytosolic forms, p44/42 mitogen-activated protein kinase (MAPK) phosphorylation, Akt kinase phosphorylation and connexin 43 phosphorylation were performed by Western blotting. Gap junction-mediated cellular communication was measured by fluorescent dye transfer. RESULTS: Treatment of WB- ras1 cells with a non-cytotoxic dose of ChK inhibited both anchorage-dependent and anchorage-independent growth. Inhibited cells were generally larger and less spindle-shaped in morphology than vehicle-treated cells, many of which were multinucleate. Removal of ChK induced cytokinesis and a return to predominantly single-nucleate cells, suggesting that ChK inhibits cytokinesis. The proportion of membrane-associated versus cytosolic forms of p21 Ras was unchanged by ChK treatment, suggesting that ChK does not act as a farnesylation inhibitor. ChK treatment decreased the level of phosphorylation of Akt kinase, a key signal transducer of the Phosphatidylinositol 3-kinase pathway. In contrast, ChK had no effect on phosphorylation of p44/42 MAPK, which mediates the MAPK/ERK Ras effector pathway. Phosphorylation of the gap junction protein, connexin 43, shown previously to increase following treatment with other anti-Ras compounds, was also not altered by ChK, which correlated with its lack of effect on gap junction-mediated cellular communication. CONCLUSIONS: Our results demonstrate that ChK inhibits Akt kinase phosphorylation and cytokinesis in ras transformed cells, which likely contribute to its ability to inhibit tumorigenic growth. PMID- 16254734 TI - Effects of unconditioned and conditioned social defeat on alcohol self administration and reinstatement of alcohol seeking in rats. AB - RATIONALE AND OBJECTIVES: We and others have shown that a stressor commonly used in laboratory studies, intermittent footshock, reinstates alcohol seeking in a rat relapse model. The effects of more ethologically relevant stressors on reinstatement have not been examined. Here, we characterized the effects of social defeat (a naturalistic stressor) or a cue associated with the defeat experience on reinstatement of alcohol seeking. We also examined the effect of unconditioned and conditioned social defeat on alcohol self-administration. METHODS: Rats were trained to self-administer alcohol (12% w/v, 1 h day(-1)), and after stable responding, one group of animals received five exposures to social defeat paired with peppermint odor prior to daily self-administration sessions. After three more self-administration sessions, these rats were tested for the effects of the peppermint odor cue on self-administration. In another group of rats, the effects of three daily exposures to social defeat paired with peppermint odor on extinction of responding were examined. After further extinction sessions, the effect of the odor cue on reinstatement was tested in these animals. The acute effect of social defeat on reinstatement was examined in another group of animals. RESULTS: Acute exposure to social defeat decreased alcohol self-administration, reduced rates of responding during extinction, and did not reinstate alcohol seeking. Exposure to a discrete odor cue previously paired with social defeat decreased alcohol self-administration but induced modest reinstatement of alcohol seeking. CONCLUSIONS: Results provide the first demonstration of reinstatement of alcohol seeking by a cue paired with social defeat and are also in agreement with previous findings on the suppressive effect of social defeat stress on alcohol self-administration. PMID- 16254735 TI - [Interventions for benign biliary strictures]. AB - Due to their potential for serious consequences, even including biliary liver cirrhosis, benign biliary strictures pose a considerable diagnostic and therapeutic challenge. In addition to inflammatory disease or an acute liver injury, iatrogenically caused biliary strictures following hepatobiliary surgery represent in 95% of cases the main cause for all benign entities. The diagnosis should be determined noninvasively with magnetic resonance cholangiopancreaticography (MRCP). Invasive techniques such as ERCP or percutaneous transhepatic cholangiography (PTC) should be reserved for unclear cases and first performed before the scheduled intervention. Depending on the site and cause of the stricture, surgical and interventional procedures are employed in the treatment of biliary strictures. The best results are obtained in short-segment strictures of the main bile duct. Interventional methods such as balloon dilation and/or stent application with concomitant drain insertion achieve patency rates of up to 75% after 5 and 55% after 12 years with a total complication rate of 5-8%. Due to the fact that most of the cases involve cicatricial fibroses, predisposition for recurrence of biliary strictures after interventional therapy can be very high, ranging up to 66% depending on the localization. PMID- 16254736 TI - Bony and cartilaginous anatomy of the patellofemoral joint. AB - The patella as the largest sesamoid bone of the human body forms the patellofemoral joint with the patellar groove of the femur. The patellofemoral joint is a complex articulation with high functional and biomechanical requirements. Several anatomical variants of both patella and the trochlea exist. Multiple clinical problems of the knee may be caused by anatomical and physiological abnormalities of this joint. Exact knowledge about the anatomy, the biomechanics and the function of the patellofemoral joint is therefore required to understand its wide range of pathology. PMID- 16254737 TI - Association of a sylvian arachnoid cyst and trigonocephaly in a developing child: importance and management. AB - INTRODUCTION: Trigonocephaly and arachnoid cysts are two different entities that may cause intracerebral pressure elevation: one by space occupying and the other by space restriction. The true incidence of association of trigonocephaly and sylvian arachnoid cyst is not known. We present a rare case of trigonocephaly associated with a sylvian arachnoid cyst in a patient that underwent forehead reconstruction and fenestration of the arachnoid cyst in the same operation. CASE REPORT: Investigation of an 11-month-old girl suffering from triangular appearance of the forehead since birth revealed metopic suture synostosis and type II sylvian arachnoid cyst. Neurological examination was normal. There were no seizure history and no other systemic malformation that could be a part of any syndromes associated with cranial synostosis. Forehead reconstruction with forehead advancement for trigonocephaly and fenestration of the arachnoid cyst to the basal cisterns were performed in the same operation. Satisfactory outcome was achieved. CONCLUSION: The coexistence of craniosynostosis and arachnoid cyst is rare. They can be treated surgically in a single session. PMID- 16254740 TI - Conduct and its consequences: attempts at debiasing jury judgments. AB - Jurors in negligence cases are supposed to judge a defendant by the reasonableness of his or her conduct and not by the consequences of that conduct. But several studies have shown that a cognitive heuristic known as hindsight bias can skew post hoc judgments of some prior behavior. Thus, jurors who must evaluate the actions of a defendant may be influenced inappropriately by the consequences of those actions. A complementary problem arises when jurors must evaluate the injuries incurred by the plaintiff. Here, jurors' knowledge about the defendant's allegedly negligent conduct can proactively influence their assessment of the plaintiff's injuries and determination of damages. The purpose of the present study was to examine the effectiveness of two procedural techniques intended to reduce or eliminate the impact of hindsight bias in negligence cases--multiple admonitions from a judge about the proper use of evidence--and bifurcation (actually withholding irrelevant evidence from jurors). We presented a re-enacted automobile negligence trial to 355 jury-eligible adults drawn from the community, varied the evidence and instructions that they heard, and measured liability judgments and damage awards from individual jurors both before and after deliberating, and from juries. Results showed that admonitions were generally ineffective in guiding jurors to the proper use of evidence but that bifurcation was relatively more effective. Deliberations had no curative effect on jurors' misapplication of evidence. PMID- 16254738 TI - Barrier method preferences and perceptions among Zimbabwean women and their partners. AB - In Zimbabwe, adult HIV prevalence is over 25% and acceptable prevention methods are urgently needed. Sixty-eight Zimbabwean women who had completed a barrier methods study and 34 of their male partners participated in focus group discussions and in-depth interviews to qualitatively explore acceptability of male condoms, female condoms and diaphragms. Most men and about half of women preferred diaphragms because they are female-controlled and do not detract from sexual pleasure or carry stigma. Unknown efficacy and reuse were concerns and some women reported feeling unclean when leaving the diaphragm in for six hours following sex. Nearly half of women and some men preferred male condoms because they are effective and limit women's exposure to semen, although they reportedly detract from sexual pleasure and carry social stigma. Female condoms were least preferred because of obviousness and partial coverage of outer-genitalia that interfered with sexual pleasure. PMID- 16254741 TI - When initial interviews are delayed a year: effect on children's 2-year recall. AB - Three- to nine-year-old children were interviewed about a medical emergency (injury requiring hospital ER treatment) two years after it occurred. Half of the number of children had been interviewed shortly after injury as well as 6 and 12 months later, while the remaining children had had only one prior interview a year after injury. There was remarkably little long-term deterioration in memory by both groups. Having a delayed initial interview had two effects, and both were relevant only to the harder-to-remember hospital treatment event: (a) The late interview group was less accurate, and (b) early-interview children had more extensive free recall, suggesting that multiple prior interviews teach children the "rules of the memory game'' when they are asked open-ended questions. Forensic implications are discussed. PMID- 16254742 TI - A problem with double-blind photospread procedures: photospread administrators use one eyewitness's confidence to influence the identification of another eyewitness. AB - In Experiment 1, photospread administrators (PAs, N = 50) showed a target-absent photospread to a confederate eyewitness (CW), who was randomly assigned to identify one photo with either high or low confidence. PAs subsequently administered the same target-absent photospread to participant eyewitnesses (PWs, N = 50), all of whom had viewed a live staged crime 1 week earlier. CWs were rated by the PAs as significantly more confident in the high-confidence condition versus low-confidence condition. More importantly, the confidence of the CW affected the identification decision of the PW. In the low-confidence condition, the photo identified by the CW was identified by the PW significantly more than the other photos; there was no significant difference in photo choice in the high confidence condition. In spite of the obvious influence exerted in the low confidence condition, observers were not able to detect bias in the photospread procedures. A second experiment was conducted to test a post-hoc explanation for the results of Experiment 1: PAs exerted influence in the low-confidence condition because they perceived the task as more difficult for the eyewitness than in the high-confidence condition. Independent observers (N = 84) rated the difficulty of the confederate's task as higher in the low-confidence condition compared with the high-confidence condition, suggesting that expectations of task difficulty might be driving the effect observed in Experiment 1. Results support recommendations for double-blind photospreads and emphasize that the same investigator should not administer photo lineups to multiple eyewitnesses in an investigation. PMID- 16254744 TI - A re-examination of the effects of biased lineup instructions in eyewitness identification. AB - A meta-analytic review of research comparing biased and unbiased instructions in eyewitness identification experiments showed an asymmetry; specifically, that biased instructions led to a large and consistent decrease in accuracy in target absent lineups, but produced inconsistent results for target-present lineups, with an average effect size near zero (Steblay, 1997). The results for target present lineups are surprising, and are inconsistent with statistical decision theories (i.e., Green & Swets, 1966). A re-examination of the relevant studies and the meta-analysis of those studies shows clear evidence that correct identification rates do increase with biased lineup instructions, and that biased witnesses make correct identifications at a rate considerably above chance. Implications for theory, as well as police procedure and policy, are discussed. PMID- 16254743 TI - Family representative payeeship and violence risk in severe mental illness. AB - Although representative payeeship is prevalent among people with mental illness and shows promise to positively influence clinically relevant outcomes, research also suggests this legal mechanism could be implemented in ways that are problematic. The current study examined whether family representative payeeship was associated with elevated risk of family violence perpetrated by persons with severe mental illness (SMI). Data were collected every 4 months for 1 year in structured interviews with N = 245 persons with SMI who received disability benefits. Multivariate analyses showed that substance abuse, history of violence, frequency of family contact, and family representative payeeship were associated with elevated odds of family violence. Analyses also showed family contact and family representative payeeship had a cumulative effect on increasing the predicted probability of family violence (controlling for covariates such as violence history and substance abuse). The data shed light on the potential for family representative payeeship to be associated with increased risk of interpersonal conflict and violence in SMI. PMID- 16254745 TI - The effect of gender in the perception of elder physical abuse in court. AB - Two experiments investigated mock jurors' perceptions of elder abuse (EA) in a physical assault case. In Experiment 1, participants read a fictional criminal trial summary of a physical assault case in which the alleged victim was 66, 76, or 86 years old. In Experiment 2, the age of the alleged victim was 76 years old, but the gender of the alleged victim and the gender of the defendant were crossed. The results of the experiments showed that women believed the alleged victim more and rendered a guilty verdict more often than men. Overall, the alleged victim was believed more than the defendant regardless of the age of the alleged victim, and most verdicts were guilty. These results are discussed in terms of the factors that affect perceptions of alleged victims of EA in court. PMID- 16254746 TI - Comparing effect sizes in follow-up studies: ROC Area, Cohen's d, and r. AB - In order to facilitate comparisons across follow-up studies that have used different measures of effect size, we provide a table of effect size equivalencies for the three most common measures: ROC area (AUC), Cohen's d, and r. We outline why AUC is the preferred measure of predictive or diagnostic accuracy in forensic psychology or psychiatry, and we urge researchers and practitioners to use numbers rather than verbal labels to characterize effect sizes. PMID- 16254747 TI - Introduction: the world challenge of work disability. PMID- 16254748 TI - Improving return to work research. AB - BACKGROUND: Despite considerable multidisciplinary research on return to work (RTW), there has been only modest progress in implementation of study results, and little change in overall rates of work disability in developed countries. METHODS: Thirty RTW researchers, representing over 20 institutions, assembled to review the current state of the art in RTW research, to identify promising areas for further development, and to provide direction for future investigations. RESULTS AND CONCLUSION: Six major themes were selected as priority areas: early risk prediction; psychosocial, behavioral and cognitive interventions; physical treatments; the challenge of implementing evidence in the workplace context; effective methods to engage multiple stakeholders; and identification of outcomes that are relevant to both RTW stakeholders and different phases of the RTW process. Understanding and preventing delayed RTW will require application of new concepts and study designs, better measures of determinants and outcomes, and more translational research. Greater stakeholder involvement and commitment, and methods to address the unique challenges of each situation are required. PMID- 16254749 TI - Prognosis and the identification of workers risking disability: research issues and directions for future research. AB - INTRODUCTION: Screening procedures based on prognostic data are an important prerequisite for prevention of disability due to low-back pain. This paper reviews the research on prognosis to delineate the most pertinent research challenges, and outlines directions for future research to improve the scientific quality and screening accuracy of prognostic efforts. METHODS: Reviews of prognosis research were examined to identify key methodological and research issues. RESULTS: Certain issues such as sampling procedures, research designs, data analyses, prognostic indicators, and follow-up procedures limit the value of prior studies. Absence of a clear conceptual framework hampers interpretation of findings and moving research questions forward. The recurrent nature of back pain and the need to effectively include the impact of employer actions and the job market were also identified as significant issues. CONCLUSIONS: Future research will be enhanced by addressing conceptual and definitional issues, applying tested and sensible measures, and careful follow-up of the study population. PMID- 16254750 TI - Integrating psychosocial and behavioral interventions to achieve optimal rehabilitation outcomes. AB - INTRODUCTION: Psychosocial factors are important contributors to work disability associated with musculoskeletal conditions. The primary objectives of this paper were 1) to describe different psychosocial interventions that have been developed to prevent prolonged work disability, and 2) to identify future research directions that might enhance the impact of programs targeting psychosocial risk factors for work disability. METHODS: Selective review of scientific literature on psychosocial and behavioral interventions and work disability. RESULTS: Most prior interventions focused on psychosocial risk factors that exist primarily within the individual (e.g., pain catastrophizing, beliefs, expectancies). Successful disability prevention will require methods to assess and target psychosocial risk factors "outside" of the individual (e.g., interpersonal conflict in the workplace, job stress, etc.) using cost-effective, multipronged approaches. Research to explore interactions among different domains of psychosocial risk factors in relation to RTW outcomes is needed. Challenges to effective secondary prevention of work disability include developing competencies to enable a range of providers to deliver interventions, standardization of psychosocial interventions, and maximizing adherence to intervention protocols. CONCLUSION: Effective secondary prevention of work disability will require research to develop cost-effective, multipronged approaches that concurrently target both worker-related and workplace psychosocial risk factors. PMID- 16254752 TI - Prevention of work disability due to musculoskeletal disorders: the challenge of implementing evidence. AB - BACKGROUND: The process of returning disabled workers to work presents numerous challenges. In spite of the growing evidence regarding work disability prevention, little uptake of this evidence has been observed. One reason for limited dissemination of evidence is the complexity of the problem, as it is subject to multiple legal, administrative, social, political, and cultural challenges. PURPOSE AND METHODS: A literature review and collection of experts' opinion is presented, on the current evidence for work disability prevention, and barriers to evidence implementation. Recommendations are presented for enhancing implementation of research results. CONCLUSION: The current evidence regarding work disability prevention shows that some clinical interventions (advice to return to modified work and graded activity programs) and some non-clinical interventions (at a service and policy/community level but not at a practice level) are effective in reducing work absenteeism. Implementation of evidence in work disability is a major challenge because intervention recommendations are often imprecise and not yet practical for immediate use, many barriers exist, and many stakeholders are involved. Future studies should involve all relevant stakeholders and aim at developing new strategies that are effective, efficient, and have a potential for successful implementation. These studies should be based upon a clearer conceptualization of the broader context and inter-relationships that determine return to work outcomes. PMID- 16254753 TI - Workplace-based return-to-work interventions: optimizing the role of stakeholders in implementation and research. AB - INTRODUCTION: The challenges of engaging and involving stakeholders in return-to work (RTW) intervention and research have not been well documented. METHODS: This article contrasts the diverse paradigms of workers, employers, insurers, labor representatives, and healthcare providers when implementing and studying workplace-based RTW interventions. RESULTS: Analysis of RTW stakeholder interests suggests that friction is inevitable; however, it is possible to encourage stakeholders to tolerate paradigm dissonance while engaging in collaborative problem solving to meet common goals. We review how specific aspects of RTW interventions can be instrumental in resolving conflicts arising from differing paradigms: calibration of stakeholders' involvement, the role of supervisors and of insurance case managers, and procedural aspects of RTW interventions. The role of the researcher in engaging stakeholders, and ethical aspects associated with that process are discussed. CONCLUSIONS: Recommendations for future research include developing methods for engaging stakeholders, determining the optimal level and timing of stakeholder involvement, expanding RTW research to more diverse work settings, and developing RTW interventions reflecting all stakeholders' interests. PMID- 16254751 TI - Physical exercise interventions to improve disability and return to work in low back pain: current insights and opportunities for improvement. AB - INTRODUCTION: There is a body of literature that indicates that physical exercise interventions, with a primary focus on improvement of functioning instead of pain relief, might be effective to stimulate return to work and improve function in workers who are absent from work due to low back pain (LBP). Successful application and implementation of these interventions however, depends on multiple factors that need to be addressed carefully in clinical practice as well as research. METHODS: Descriptive literature review, to identify an overview of current knowledge with respect to the safety, content- and context-related aspects of physical exercise interventions, issues relating to timing, the influence of treatment confidence and patient expectations, and the process of changing provider and employer behavior. RESULTS: Physical exercises are not associated with an increased risk for recurrences. The effects of interventions may vary depending on content-related factors (i.e., type of exercises, dosage, frequency, skills of the healthcare providers, etc.) and contextual factors (i.e., treatment setting, compensation system, etc.). Treatment confidence and patients' expectations also significantly influence outcomes of physical exercise interventions. Timing is also important; interventions targeting return to work, applied during the acute phase of work absenteeism, compete with a high rate of spontaneous recovery and may therefore be inefficient. CONCLUSIONS: Despite numerous studies, more quantitative and qualitative investigations are needed to further clarify the requirements for a successful application and implementation of physical exercise interventions for disabled workers with low back pain. PMID- 16254754 TI - Return-to-work outcomes following work disability: stakeholder motivations, interests and concerns. AB - INTRODUCTION: Satisfaction with return-to-work (RTW) outcomes is dependent on many factors, including a clear exposition of what people define as a "good outcome" and the information they use to determine if such an outcome has been achieved. This paper defines the key stakeholders involved in the RTW process and discusses the need to understand their motivations, interests, and concerns. METHODS: A review of the literature and discussions with RTW researchers conducted by a multidisciplinary group of academic researchers. RESULTS: Our analysis suggests that RTW stakeholders can share the goal of a successful RTW; however, this consensus has to be viewed in light of other, sometimes competing, goals and the environments in which stakeholders operate. CONCLUSIONS: It is suggested that more clearly articulating and operationalizing stakeholders' perspectives will allow researchers to advance the understanding of RTW interventions and outcomes. PMID- 16254755 TI - A developmental conceptualization of return to work. AB - INTRODUCTION: Although return to work (RTW) is a phenomenon that has been researched for many years, our ability to understand and improve outcomes is still limited. As an avenue for advancing the field, this paper presents an alternative way of thinking about RTW. METHOD: The conceptualization was constructed based on a review of the literature and the comments of RTW and workers' compensation researchers. RESULTS: RTW is presented as an evolving process, comprising four key phases: i.e., "off work," "work re-entry," "retention," and "advancement." In addition, multiple phase-specific outcomes that may be used to evaluate RTW success are advanced. CONCLUSION: Broadening thinking about RTW to take into consideration the complexities of its developmental nature holds promise for understanding and improving RTW, as it not only clarifies the importance of incremental milestones, but also facilitates intervention choice and evaluation. PMID- 16254756 TI - The effects of a graded activity intervention for low back pain in occupational health on sick leave, functional status and pain: 12-month results of a randomized controlled trial. AB - INTRODUCTION: Behaviorally oriented graded activity interventions have been suggested for sick-listed workers with low back pain on return to work, but have not been extensively evaluated. METHODS: One hundred and thirty-four workers were randomly assigned to either a graded activity intervention (n = 67) or usual care (n = 67) and followed-up for 12 months. RESULTS: The graded activity group returned back to work faster with a median of 54 days compared to 67 days in the usual care group. The graded activity intervention was more effective after approximately 50 days post-randomization (HRR = 1.9, CI = 1.2-3.1, p = 0.01). Differences between the groups in number of recurrent episodes, total number of days of sick leave due to low back pain, and total number of days of sick leave due to all diagnoses, were in favor of the graded activity group, although not statistically significant. No effects of the graded activity intervention were found for functional status or pain. CONCLUSION: Graded activity intervention is a valuable strategy to enhance short-term return to work outcomes. PMID- 16254757 TI - Interorganizational collaboration in occupational rehabilitation: perceptions of an interdisciplinary rehabilitation team. AB - INTRODUCTION: Various obstacles to and facilitators of collaboration between an interdisciplinary work rehabilitation team and the stakeholders (workers, insurers, physicians, and employers) exist, but are not well characterized. METHODS: An observational study was conducted, using videotapes of interdisciplinary team discussions of ongoing cases involving 22 workers absent from work due to musculoskeletal disorders. The actions taken and strategies adopted by the team in an effort to overcome the obstacles to collaboration were studied. RESULTS: Various factors influence collaboration between the rehabilitation team and the stakeholders. In general, stakeholder endorsement of the team's therapeutic principles and confidence in their approach emerged as particularly important factors. Diverse strategies, most often, education and awareness-raising, were used by the team to foster collaboration among the parties. CONCLUSIONS: This study provides greater insight into the factors affecting collaboration among a rehabilitation team, an injured worker and other stakeholders. The results may improve understanding of the actions taken by rehabilitation teams and help to optimize their practices. PMID- 16254758 TI - Prognostic factors for duration of sick leave due to low-back pain in dutch health care professionals. AB - BACKGROUND: Information on prognostic factors for duration of sick leave due to low-back pain (LBP) is growing. In this prospective cohort study prognostic factors for duration of sick leave and course of disability were identified in a very early stage of sick leave due to LBP in an occupational health care setting. METHODS: A total of 615 workers calling in sick from work due to LBP at an occupational health service (OHS) of one institution completed a questionnaire. Duration of follow-up was 26 weeks. Prognostic factors were identified by means of Cox regression analysis and multiple linear regression analysis, and explained variance was calculated. RESULTS: Median time to first return to work (RTW) was 5 days (Inter Quartile Range (IQR) = 2-12). Median time to lasting return to work (LRTW) was 6 days (IQR = 3-13). The final model for delayed RTW included self reported expected duration of sick-leave, treatment by GP or specialist, care seeking, diminished mobility, and the interaction between care-seeking and diminished mobility. The final model for LRTW included: self-reported expected duration of sick-leave, treatment by GP or specialist, care-seeking, the interaction between the self-reported expected duration of sick-leave and seeking care at OHS, complaints due to job stress, diminished mobility, and the interactions between expected duration of more then 10 days and seeking OP care and diminished mobility. Median total days on sick leave (TDSL) was 6 days (IQR = 3-13 days). The final model for TDSL included: age, expected duration of sick leave, treatment by GP or medical specialist, seeking OP care, complaints due to physical load, and diminished mobility. Explained variance (R2) of these models ranged from 30 to 35%. CONCLUSIONS: Poor prognosis for duration of sick leave can be identified by means of a simple questionnaire administered on the first day of sick leave. Workers at high risk for longer duration of sick leave (all outcomes) expected to stay off work longer, were already being treated by a MD, sought OP care, and had diminished mobility. In case of delayed LRTW workers reported job stress as a possible cause of sick leave. Interactions were found in both RTW and LRTW between care-seeking and diminished mobility and in LRTW between expected duration of sick leave and seeking OP care. Older age increased TDSL with borderline significance. PMID- 16254759 TI - Workplace-based return-to-work interventions: a systematic review of the quantitative literature. AB - INTRODUCTION: A systematic review was conducted to review the effectiveness of workplace-based return-to-work (RTW) interventions. METHOD: Seven databases were searched, in English and French, between January 1990 and December 2003 for peer reviewed studies of RTW interventions provided at the workplace to workers with work disability associated with musculoskeletal or other pain-related conditions. Methodological quality appraisal and data extraction were conducted by pairs of reviewers. RESULTS: Of a total of 4124 papers identified by the search, 10 studies were of sufficient quality to be included in the review. There was strong evidence that work disability duration is significantly reduced by work accommodation offers and contact between healthcare provider and workplace; and moderate evidence that it is reduced by interventions which include early contact with worker by workplace, ergonomic work site visits, and presence of a RTW coordinator. For these five intervention components, there was moderate evidence that they reduce costs associated with work disability duration. Evidence for sustainability of these effects was insufficient or limited. Evidence regarding the impact of supernumerary replacements was insufficient. Evidence levels regarding the impact of the intervention components on quality-of-life was insufficient or mixed. CONCLUSIONS: Our systematic review provides the evidence base supporting that workplace-based RTW interventions can reduce work disability duration and associated costs, however the evidence regarding their impact on quality-of-life outcomes was much weaker. PMID- 16254760 TI - Soil bacterial diversity responses to root colonization by an ectomycorrhizal fungus are not root-growth-dependent. AB - The hypothesis tested in this present study was that the ectomycorrhizosphere effect on the bacterial community was not root-growth-dependent. The impacts of ectomycorrhizal infection (Pisolithus albus COI007) and a chemical fertilization to reproduce the fungal effect on root growth were examined on (1) the structure of bacterial community and (2) fluorescent pseudomonad and actinomycete populations in the mycorrhizosphere of Acacia auriculiformis using both culture independent and culture-dependent methods. A. auriculiformis plants were grown in disinfested soil in pots with or without addition of the ectomycorrhizal fungus or N/P/K fertilization (to reproduce the fungal effect on root growth) for 4 months and then transferred to 20-L pots filled with nondisinfested sandy soil. The fungal and fertilizer applications significantly improved the plant growth after 4-month culture in the disinfested soil. In the nondisinfested cultural substrate, these positive effects on plant growth were maintained. The total soil microbiota was significantly different within the treatments as revealed from DNA analysis [denaturing gradient gel electrophoresis (DGGE)]. The structure of fluorescent pseudomonad populations was also affected by fungal and fertilizer applications. In contrast, no qualitative effect was observed for the actinomycete communities within each treatment, but fungal inoculation significantly decreased the number of actinomycetes compared to the fertilizer application treatment. These results show that the mycorrhizosphere effect is not root-growth-dependent but is mainly due to the presence of the ectomycorrhizal fungus and more particularly to the extramatrical mycelium. PMID- 16254761 TI - Comparison of 16S rRNA and 16S rDNA T-RFLP approaches to study bacterial communities in soil microcosms treated with chromate as perturbing agent. AB - Transcripts of ribosomal RNA have been used for assessing the structure and dynamics of active bacterial populations; however, it remains unclear whether the information provided by community profiling derived from RNA is different from that derived from DNA, particularly when a selective pressure is applied on the bacterial community. In the present work, terminal-restriction fragment length polymorphism (T-RFLP) community profiles based on DNA and RNA extracted from soil microcosms treated with a toxic concentration of chromate were compared. Microcosms of a nonpolluted agricultural soil and of a heavy-metal-rich soil (serpentine) were treated with chromate and DNA and RNA were extracted. T-RFLP analysis was performed on amplified and retro-amplified 16SrRNA gene sequences, and band profiles obtained from samples of DNA and of RNA were compared. Some of the T-RFLP bands, identified as peculiar peaks in the profiles, were cloned and sequenced for taxonomic interpretation. Results indicated that: (1) community profiles derived from RNA and DNA were partly overlapping; (2) there was a strong correlation between the dynamics shown by RNA- and DNA-based T-RFLP profiles; (3) chromate addition exerted a clear effect on both agricultural and serpentine soil bacterial communities, either at the DNA and at the RNA level; however, the profiles derived from RNA showed sharper differences between treated and control samples than that of DNA-based profiles. PMID- 16254762 TI - Distribution of Melissococcus plutonius in honeybee colonies with and without symptoms of European foulbrood. AB - A sensitive hemi-nested polymerase chain reaction (PCR) was used for detection of Melissococcus plutonius, the causative agent of European foulbrood (EFB). Sampling was made in Switzerland, where EFB is a widespread disease and incidences have increased in recent years. Larvae from brood samples with and without clinical signs of disease (n=92) and honey (n=92) from the same colonies were investigated. Individual larvae (n=60) and pupae (n=30) from diseased brood in single colonies were also investigated to study the distribution of the bacterium within the brood between larvae. M. plutonius was detected in larvae in all apiaries where symptoms of EFB could be seen, but not in all colonies judged as cases of EFB in the field, when healthy-looking larvae from such colonies were tested. The occurrence of the bacterium within the brood was not limited to larvae with symptoms only, but was mainly found in diseased larvae. The bacterium was also found in pupae. Healthy-looking larvae-even from heavily diseased combs failed, in a number of cases, to amplify product in the PCR. M. plutonius could only be detected in 35% of the brood nest honey from clinically diseased colonies. PMID- 16254764 TI - The Self-Efficacy Questionnaire for Depression in Adolescents (SEQ-DA). Development and psychometric evaluation. AB - The 12-item clinician or self-administered Self-Efficacy Questionnaire for Depression in Adolescents (SEQ-DA) was developed as a measure of perceived ability to cope with depressive symptomatology. This study examined the reliability and validity of the SEQ-DA in a clinical population of 130 adolescents that were receiving treatment for depression. Psychometric evaluation revealed good internal consistency and test-retest reliability. Results indicated that higher SEQ-DA scores were associated with lower self-rated depression scores (Reynolds Adolescent Depression Scale), which is evidence of good construct validity. Further, higher SEQ-DA scores prior to treatment predicted better outcome at the end of the 3 months of treatment and at 6 months post-treatment. Therefore, the SEQ-DA has a potentially useful role in clinical work and research with depressed young people. PMID- 16254763 TI - Poly(ethylene oxide)-modified poly(beta-amino ester) nanoparticles as a pH sensitive system for tumor-targeted delivery of hydrophobic drugs: part 2. In vivo distribution and tumor localization studies. AB - PURPOSE: This study was carried out to determine the biodistribution profiles and tumor localization potential of poly(ethylene oxide) (PEO)-modified poly(beta amino ester) (PbAE) as a novel, pH-sensitive biodegradable polymeric nanoparticulate system for tumor-targeted drug delivery. METHODS: The biodistribution studies of PEO-modified PbAE and PEO-modified poly(epsilon caprolactone) (PCL), a non-pH-sensitive polymer, nanoparticle systems were carried out in normal mice using 111indium-oxine [111In] as a lipophilic radiolabel encapsulated within the polymeric matrix, and the distribution of the nanoparticles was studied in plasma and all the vital organs following intravenous administration. Solid tumors were developed on nude mice using human ovarian carcinoma xenograft (SKOV-3) and the change in concentrations of tritium [3H]-labeled paclitaxel encapsulated in polymeric nanoparticles was examined in blood, tumor mass, and liver. RESULTS: Study in normal mice with a gamma-emitting isotope [111In] provided a thorough biodistribution analysis of the PEO-modified nanoparticulate carrier systems, whereas 3H-paclitaxel was useful to understand the change in concentration and tumor localization of anticancer compound directly in major sites of distribution. Both PEO-PbAE and PEO-PCL nanoparticles showed long systemic circulating properties by virtue of surface modification with PEO-containing triblock block copolymer (Pluronic stabilizer. Although the PCL nanoparticles showed higher uptake by the reticuloendothelial system, the PbAE nanoparticles effectively delivered the encapsulated payload into the tumor mass. CONCLUSIONS: PEO-modified PbAE nanoparticles showed considerable passive tumor targeting potential in early stages of biodistribution via the enhanced permeation and retention (EPR) mechanism. This prompts a detailed biodistribution profiling of the nanocarrier for prolonged periods to provide conclusive evidence for superiority of the delivery system. PMID- 16254765 TI - Psychopathology in children: improvement of quality of life without psychiatric symptom reduction? AB - OBJECTIVE: The aim of this study was to assess the association between change in psychopathology and Quality of Life (QoL) across time in children with high levels of psychopathology. METHODS: A referred sample of 126 seven- to 19-year olds was studied across a 1-year follow-up period. Information concerning QoL and psychopathology was obtained from parents. RESULTS: Overall, 38.1% of children showed neither psychiatric symptom reduction nor QoL improvement, 33.3% of children showed both a clinically significant psychiatric symptom reduction and QoL improvement, and 28.6% of children showed either psychiatric symptom reduction or QoL improvement. In 11.1% of all children, QoL improved, while the level of psychopathology remained high. Age, gender, or psychiatric diagnosis did not predict a poor outcome of persistently high psychopathology scores and poor QoL. CONCLUSION: QoL in children with psychiatric problems may be improved by reducing psychiatric symptoms in a number of children, but it is also possible to improve QoL without psychiatric symptom reduction. This implicates that QoL should become an important aim and treatment outcome measure of psychiatric treatment programs, especially since psychopathology tends to persist. PMID- 16254766 TI - Bodily pain and associated mental distress among immigrant adolescents. A population-based cross-sectional study. AB - OBJECTIVE: The aim of this study was to describe differences among immigrant groups in bodily pain, and analyze its association with mental distress. METHOD: A population-based cross-sectional study was carried out involving tenth grade pupils in Oslo. Of the 7,343 pupils that participated, one-quarter were first- or second-generation immigrants. The Hopkins Symptom Checklist-10 was used to measure mental distress. All information on pain and mental distress was self reported. RESULTS: Girls reported more bodily pain from all types of pain. Headache was the most prevalent pain site across gender and immigrant groups. Strong associations between mental distress and number of pain sites were found for all immigrant groups. Neck and shoulder pain yielded the highest odds ratio (OR) for mental distress among the majority of the immigrant groups. The Sub Saharan African group had the highest adjusted OR for mental distress [OR=9.8 (1.1-82.7)] when reporting three or more pain sites, and the Indian Subcontinent the lowest [OR=4.0 (1.8-8.8)]. CONCLUSION: The differences in number and types of pain were small, though significant between the different immigrant groups. Adolescents from Sub-Saharan Africa seem to react with more mental distress to bodily pain than adolescents emigrating from the Indian Subcontinent. PMID- 16254767 TI - Why do adolescents with bulimia nervosa choose not to involve their parents in treatment? AB - BACKGROUND: Although the use of family therapy for adolescents with anorexia nervosa is well established, there has been limited research into the efficacy of family therapy in adolescents with bulimia nervosa (BN). No previous research has investigated why individuals with BN do or do not involve their parents in treatment. This is an exploratory study aimed at determining whether there are any differences between these individuals in terms of eating disorder symptomatology, psychopathology, familial risk factors, patients' perception of parental expressed emotion (EE) and family functioning. METHODS: Participants were 85 adolescents with BN or Eating Disorder Not Otherwise Specified, recruited to a randomised controlled evaluation of the cost-effectiveness of cognitive behavioural guided self-care vs. family therapy. Participants were interviewed regarding the history of their eating disorder and completed self-report measures. RESULTS: Patients who did not involve their parents in treatment were significantly older, had more chronic eating disorder symptoms, exhibited more co morbid and impulsive behaviours and rated their mothers higher in EE. However, they did not have more severe eating disorder symptomatology. CONCLUSIONS: These preliminary findings, although in need of replication with a larger sample and limited by the attrition rate in some of the self-report measures, indicate that patients who did not involve their parents in treatment may perceive their mothers as having a more blaming and negative attitude towards the patient's illness. Public awareness about BN needs to be raised, focusing on reducing the stigma and negative views attached to this illness. PMID- 16254768 TI - Mental health, behaviour problems and incidence of child abuse at the age of 16 years. A prospective longitudinal study of children born at psychosocial risk. AB - Of the 1,575 pregnant women registered at the public Antenatal Health Care Service in the city of Linkoping, Sweden during 1983, an index group of 78 women was identified that met specific well-defined psychosocial risk criteria related to drug addiction, mental insufficiency, and particular social circumstances of possible relevance to problems of pregnancy and early child development. A further 78 pregnant women who did not meet the inclusion criteria were used as a reference group. The present study is a 16-year follow-up in which 43 (57%) of the original index children and 63 (82%) of the original reference children were examined on indices of mental health, and the presence of child abuse. Their mental health was assessed with the Child Behaviour Checklist (CBCL) completed by the mothers and the Youth Self-Report (YSR) completed by the adolescents. The incidence of child abuse and Social Welfare interventions was obtained from Social Welfare records. The index children, especially the boys, displayed significantly poorer mental health as assessed by both CBCL (p<0.05) and YSR (p<0.02). Being an index child increased the odds ratio 16-27 times for different Social Welfare interventions, and child abuse had been investigated in 27% of the index children compared to 1% of the reference children. PMID- 16254769 TI - Girls with anorexia nervosa as young adults. Self-reported and parent-reported emotional and behavioural problems compared with siblings. AB - This follow-up study had three objectives: 1) to investigate emotional and behavioural problems, adaptive functioning and substance use in former anorexia nervosa (AN) patients compared with siblings, 2) to compare information obtained from different informants, and 3) to compare questionnaire results with interview results. Fifty (of 55) female AN patients, representative for AN patients under 18 years referred to county health services, were assessed at a mean of 8.8 years after treatment start with the Young Adult Self-Report and the Young Adult Behaviour Checklist (mean age 23.1 years). In all, 48 patients, 25 siblings, 33 mothers and 27 fathers participated in the questionnaire study. Although 41/50 (82 %) had recovered from their eating disorder, the former AN patients had substantially more self-reported and parent-reported problems than their siblings, particularly with regard to Internalising Problems and on the Anxious/Depressed syndrome scale. Cross-informant agreement between the parents and between parents and patients was high, but low between parents and siblings. The patients with psychiatric diagnoses at follow-up had substantially higher problem scores than those without diagnoses both on the self-report and the parent-report, supporting the validity of the questionnaires. In conclusion, the self- and parent-reports showed a high level of Internalising Problems and were useful instruments in the assessment of former AN patients. PMID- 16254770 TI - The Maudsley long-term follow-up of child and adolescent depression. Predicting costs in adulthood. AB - BACKGROUND: Little is known about the long-term economic impact of childhood depression. The aim of this study is to identify and examine the links between the characteristics of children with depression and the costs of services used in adulthood. METHODS: Subjects (N=149) who had attended psychiatric services in South London for depression were followed up on average 20.7 years later. Sociodemographic and illness characteristics were recorded in childhood and service use in adulthood was measured. Costs were calculated and multiple regression models were developed to explain variations in cost, with a comparison between ordinary least squares estimation and a generalised linear model. RESULTS: Service use and cost data were available on 140 subjects. The mean annual cost was 890 pounds Sterling (range 0 pounds Sterling-7532 pounds Sterling). Predictors of cost variations in both models were age at initial referral, level of childhood anxiety, and the presence of comorbid conduct disorder. In the ordinary least squares model, a family history of psychiatric illness was inversely related to cost, whilst in the generalised linear model there was an inverse link between peer/sibling relationship problems and costs. The models could explain 24 % and 20% of cost variation, respectively. CONCLUSIONS: It is possible to explain a reasonable amount of variation in adult service costs from factors describing the characteristics of children at the time of receiving care. PMID- 16254773 TI - American Chemical Society--230th National Meeting. Technical achievements in organic chemistry. PMID- 16254772 TI - American Chemical Society--230th National Meeting. Medicinal chemistry. PMID- 16254774 TI - American Chemical Society--230th National Meeting. Proteins, peptides, amino acids and enzyme inhibitors. PMID- 16254775 TI - American Chemical Society 230th National Meeting. Pain, anxiety and depression. PMID- 16254776 TI - American Chemical Society 230th National Meeting. Hormone receptor modulators and antiviral agents. PMID- 16254777 TI - American Chemical Society 230th National Meeting. Neurological disorders. PMID- 16254778 TI - American Chemical Society 230th National Meeting. Highlights. PMID- 16254779 TI - Tumor progression & therapeutic resistance--Second International Conference. PMID- 16254780 TI - AIDS Vaccine 2005. PMID- 16254781 TI - Medicinal Chemistry--13th RSC-SCI Symposium. PMID- 16254782 TI - The International Society on Thrombosis and Haemostasis--XXth Annual Congress. PMID- 16254783 TI - The European Partnering and Investment Conference. PMID- 16254784 TI - The future of G protein-coupled receptors as targets in drug discovery. AB - G protein-coupled receptors (GPCRs) represent the most abundant drug targets today. A large number of GPCR-based drugs have already been developed for a variety of indications in human disease. However, orphan receptors with unidentified ligands serve as potential targets still to be explored. Moreover, research on the interaction of GPCRs with different molecules in the signal transduction pathways, and further studies on receptor dimerization may also lead to the discovery of new drugs. Structure-based drug design will eventually play a key role in generating better and more selective drugs more rapidly when high resolution structures of GPCRs can be provided by expression, purification and crystallography technologies. PMID- 16254785 TI - Therapeutic switching: a new strategic approach to enhance R&D productivity. AB - New chemical entity discovery and development is a long, expensive and risky way of producing new therapeutics. An alternative approach, namely finding new uses for existing drugs, offers substantial advantages in terms of cost, time and risk, and represents a highly attractive strategy to drug discovery. Patents protecting the new use may also be obtained. There are multiple different classes of such R and D programs, depending on whether the original development is still protected by composition of matter patents, and whether the compound was ever fully developed/marketed for its original indication. There are also shortened R and D programs for new uses for stereoisomers and metabolites of existing drugs. Case histories in all areas are presented. PMID- 16254786 TI - The emergence of chemical genomics in drug discovery. AB - The interaction of small organic molecules with proteins and other macromolecules is fundamental to drug action. Chemical genomics employs a combination of chemistry, genomics and informatics to study these drug-target interactions in a systematic and global manner in order to improve the efficiency of the drug discovery process. PMID- 16254787 TI - High-throughput screening using genome-wide siRNA libraries. AB - Small-interfering RNA (siRNA) technology is a new approach for blocking the production of a protein by specific degradation of the messenger RNA that encodes the protein. The high potency of siRNA and the relative ease of application have led to broad implementation of the method in many aspects of the life science research. The combination of the developments in its application and in siRNA libraries to cover whole genomes has further enabled genome-wide high-throughput functional screening. PMID- 16254788 TI - Tesaglitazar. AB - AstraZeneca plc is developing tesaglitazar, an oral dual peroxisome proliferator activated receptor alpha/gamma agonist, for the potential improvement of dyslipidemia and glycemic control in type 2 diabetic patients. PMID- 16254789 TI - Inflammation 2005 - Seventh World Congress. Respiratory inflammation. AB - In this conference, unlike previous conferences in this series, relatively little time was given to new drug therapy approaches for the treatment of inflammation. Instead, the emphasis was on basic mechanisms with a strong bias toward Australian research. The etiology of chronic obstructive pulmonary disease (COPD) and asthma was discussed in detail with an intensive consideration of mechanisms involved in the development of corticosteroid resistance, particularly the role of histone deacetylase. Various rodent model systems for testing potential inhibitors of COPD and asthma were also presented. PMID- 16254790 TI - Inflammation 2005 - Seventh World Congress. Rheumatoid arthritis and cell signaling. AB - Novel targets for antirheumatic therapy were discussed including synoviocyte CD97 and the T-cell co-receptor CTLA-4: an immunoglobulin fusion protein of CTLA-4 (abatacept (Bristol-Myers Squibb Co)) has recently been revealed to be a well tolerated and effective therapeutic agent for rheumatoid arthritis. Furthermore, cell-signaling molecules were a major topic, and cytoplasmic adaptors of toll like receptor responses, the kinases JNK and Lyn and the macrophage surface molecule CD200 were considered as potential targets for novel therapies. PMID- 16254791 TI - Inflammation 2005 - Seventh World Congress. Highlights I. PMID- 16254792 TI - Inflammation 2005 - Seventh World Congress. Highlights II. PMID- 16254793 TI - Drug Discovery Technology and Development 2005 - IBC's Tenth World Congress. Protein-protein interactions, biomarkers and RNAi. PMID- 16254794 TI - Drug Discovery Technology and Development 2005 - IBC's Tenth World Congress. Emerging and early stage company showcase pre-conference symposium. PMID- 16254795 TI - Drug Discovery Technology and Development 2005 - IBC's Tenth World Congress. Cancer mouse models and other techniques for improved prediction of efficacy. PMID- 16254796 TI - Drug Discovery Technology and Development 2005 - IBC's Tenth World Congress. Filling the gaps in compound libraries and enhancing compound collections. PMID- 16254798 TI - Biotechnology - 12th European Congress. PMID- 16254797 TI - Anticancer Drug Discovery and Development - SRI's Seventh Annual Summit. PMID- 16254799 TI - HIV Pathogenesis and Treatment - Third International AIDS Society Conference. PMID- 16254801 TI - Cutting through the obstacles and resurrecting the promise of gene therapy. AB - Gene therapy is a promising biomedical discipline that could potentially lead to new treatments and perhaps long-term curative effects for a plethora of diseases including hereditary disorders, cardiovascular and neurological diseases, cancer, diabetes and even infectious or autoimmune diseases. These diseases affect millions of people worldwide and the development of effective and safe gene-based drugs obviously represent a tremendous market potential. Convincing evidence continues to emerge from clinical trials demonstrating that gene therapy can be effective in patients suffering from a limited number of different diseases. Nevertheless, as with any emerging new biomedical discipline, gene therapy has also faced a number of setbacks, and there have been concerns regarding the safety of some gene delivery approaches, however, these hurdles are not insurmountable. Gene transfer technologies are improving rapidly and have led to the development of new and more efficacious gene delivery approaches with fewer side effects. The success of gene therapy is still highly dependent upon the continuous development of improved gene delivery technologies, the progress of which should hopefully and ultimately cure diseases that are refractory to current treatment paradigms. PMID- 16254800 TI - The European Association of Addiction Therapy - Inaugural Conference. PMID- 16254802 TI - Using computational modeling to drive the development of targeted therapeutics. AB - Computational biology is impacting drug design and has the potential to revolutionize the drug development process. Here, we describe how data-driven computational models can optimize the design of targeted therapeutics to yield improved safety and efficacy. We propose that, in the future, this approach will improve drug discovery, clinical development, diagnosis and treatment. PMID- 16254803 TI - Challenges and opportunities for positron-emission tomography in personalized medicine. AB - Of the non-invasive functional imaging tools available, positron-emission tomography (PET) is generally expected to have the greatest potential for delivering the vision of personalized medicine. This can be achieved by the production of designer PET probes with exquisite sensitivity that can profile key biological processes that are specific to a disease. However, the challenge for the PET field will be its capability to produce and supply cost-effective PET probes to the wider community. This will most likely be achieved through the provision of long-lived PET radioisotopes with imaging qualities that match the performance of the evolving PET camera technology and chemistry that is amenable to kit formulations. Additionally, of the emerging PET radiotracers, (64)Cu has the optimum chemistry and emission characteristics for the wider application of PET in personalized medicine. PMID- 16254804 TI - The impact of computing technology on pharmaceutical and biotech research. AB - Pharmaceutical and biotechnology companies continue to make investments in research techniques including those using high-performance computing technology. Much of this research is fueled by the need to analyze the ever-growing store of genomic and proteomic data. However, many research and development investments, including techniques to mine newly produced genomic data, have not provided the anticipated improvements in productivity. This raises the questions of whether advancements expected from next-generation computing technology can translate into tangible benefits for pharmaceutical and biotechnology researchers, and whether researchers can capitalize on an increased understanding of biological systems using tools made more readily available in high-performance computing. The author provides some background on the source of the advancements in the computing industry and offers examples from other scientific applications that point to potential benefits in life science research. PMID- 16254805 TI - Becatecarin (Helsinn Healthcare). AB - Helsinn Healthcare SA is developing becatecarin, a topoisomerase II inhibitor for the potential intravenous treatment of cancer. PMID- 16254806 TI - Intraoperative imaging of the brachial plexus by high-resolution ultrasound. AB - The authors attempted to perform direct high resolution ultrasound imaging of the intraneural ultrastructure of the brachial plexus for intraoperative image guidance in brachial plexus surgery. The brachial plexuses of four fresh-frozen cadaver specimens were surgically exposed in a standard approach used in routine brachial plexus surgery to undergo direct ultrasound examination using a 15 MHz SonoCT scanhead. By placing the scanhead directly onto the epineurium, all components of the brachial plexus were directly visualized in an axial plane and compared with histologic findings. The internal neural structure at different levels could be visualized in high resolution, showing the specific fascicular pattern. The histologic processing revealed good correlation with the intraneural topography demonstrated on ultrasound. In the study, high resolution ultrasound examination of the brachial plexus showed substantial details of the ultrastructure of neural tissue, which may play a role in intraoperative image guidance in the surgical treatment of patients with brachial plexopathy. PMID- 16254807 TI - Lacerations to the brachial plexus: surgical techniques and outcomes. AB - The charts of patients with 201 brachial plexus elements sustaining operative lacerations and managed at Louisiana State University Health Sciences Center (LSUHSC) were reviewed retrospectively. Results for elements injured by sharp transections and undergoing suture repairs performed within 72 hr, as well as secondary suture and secondary graft repairs are documented. Similarly, results for secondary end-to-end suture anastomosis and secondary graft repairs for elements sustaining blunt transections are reviewed. Results for neurolysis, end to-end suture anastomosis, and graft repairs for plexus elements in continuity despite the laceration injury are reviewed. Outcomes for the LSUHSC series of brachial plexus lacerations are one of the best of all LSUHSC plexus injuries, even for elements generally viewed as unfavorable for repair. Lesions in continuity with positive nerve action potentials (NAPs) had the best outcomes. PMID- 16254808 TI - Combined thigh flap for closure of massive trunk defect. AB - A massive trunk defect resulting from resection of recurrent sarcoma was reconstructed with a combined free flap incorporating medial, anterior, and lateral thigh tissues. This flap included the tensor fasciae latae, lateral thigh perforator, and rectus femoris, all based on the lateral femoral circumflex pedicle. A saphenous vein conduit enabled this flap to replace resected tissues at the lower thorax. Combining the three different commonly used thigh flaps on a single large pedicle enabled transfer of a 47.5 x 33.5-cm mega-flap. PMID- 16254809 TI - Microdissection thinning of a pedicled deep inferior epigastric perforator flap for burn scar contracture of the groin: case report. AB - A severe burn scar contracture of the groin in a 25-year-old woman was successfully reconstructed with microdissection and thinning of a pedicled deep inferior epigastric perforator (DIEP) flap. This perforator flap is a relatively thin cutaneous flap, but may still require secondary defatting procedures, especially when used for coverage of mobile anatomic areas such as the groin and axilla. A deep inferior epigastric perforator flap was radically thinned by excising the subcutaneous fatty layer very carefully under loupe magnification, to preserve the subdermal vascular plexus. One year after surgery, the patient can abduct and rotate the hip freely without limitation. PMID- 16254811 TI - Toe-to-hand transfer from a cross-foot replantation in a traumatic four-extremity amputation. AB - Treatment of mutilating hand injuries often requires multiple innovative procedures. This report describes a case of multi-limbed amputations from a train injury and the extraordinary microsurgical approaches for a two-hand reconstruction. The first stage of the procedure was a cross replantation of the left foot to the right leg. The second stage was a combined second and third toe transfer from the cross replantation of the left foot to the right hand, and a sensate fibular osteocutaneous flap transfer for left hand reconstruction. Satisfactory function was restored, including good protective sensation. These kinds of extraordinary microsurgical approaches are useful salvage procedures for hand reconstruction, when presented with a case of multi-limb amputation. PMID- 16254813 TI - Cutaneous perforators of the upper arm and clinical applications. AB - The authors investigated the distribution of constant cutaneous perforators in the upper arm. A total of 20 amputated upper arms of 10 fresh Korean cadavers were used for the study. Red latex was injected into the axillary arteries of ten specimens and lead oxide-gelatin mixture (radiopaque material) in the other ten. The cutaneous perforators were then identified by dissection and radiography. The upper arm had several (range: 5.7 to 6.3) perforating arteries in the subfascial plane, but only four fasciocutaneous perforators were constant: one in the medial intermuscular septum and three in the lateral intermuscular septum. The constant medial perforators were included in a circle of 2.89 cm in diameter, the center of which was 8.9 cm above and 1.2 cm medial to the medial epicondyle. The mean length and diameter of the extended pedicle of the medial perforator was 2.78 cm and 0.94 mm, respectively. The lowermost constant lateral perforators were included in a circle of 2.44 cm in diameter, the center of which was 16.8 cm above and 0.5 cm medial to the lateral epicondyle. The mean length and diameter of the extended pedicle of the lateral perforator was 2.88 cm and 0.84 mm, respectively. A flap based on the perforator of the medial intermuscular septum is not as simple as with the lateral intermuscular perforator, but direct closure of the donor site is more favorable. It is safe to design a free skin flap with knowledge of its dominant perforator. PMID- 16254814 TI - Functional recovery of rat hind-limb allografts. AB - Few papers have assessed the long-term functional recovery of animal limb allografts. In this study, the functional recovery of rat limb allografts was serially and quantitatively investigated for a period of 1 year. The donor's hind limb was orthotopically transplanted into the recipient. Fifteen recipients with allografts were treated with FK506. Functional recovery of the grafted limb was assessed serially by cutaneous reaction test, walking track analysis, and electrophysiologic evaluation. Sensibility improved to a similar extent in both isografts and allografts, and the recovery rate at 1 year was 68 percent, compared to the normal side. Sciatic function index significantly improved to - 70 points after 1 year. The amplitude recorded from the gastrocnemius muscle significantly improved, and the ratio compared to the normal side was 43 percent. Limb isografts and allografts treated with FK506 showed no significant differences in functional recovery. The data can be used as a reference standard for future investigations. PMID- 16254815 TI - Effect of VEGF on tail artery interpositional loop (TAIL) flap: a rodent model for flap prefabrication. AB - The authors introduce an experimental model of flap prefabrication, the tail artery interpositional loop (TAIL) flap. In this model, an arterial segment from a rat tail is used to create an arteriovenous (A-V) fistula. This fistula is positioned beneath the abdominal skin flap to vascularize the overlying tissue, and a barrier of Silastic sheeting is placed below the fistula to prevent vascular ingrowth from the underlying bed. The efficacy of this new model was tested by investigating the effect of a single topical application of recombinant human VEGF (165). Treatment and control groups each contained 20 animals. In the control group, mean survival skin areas at 1, 2, 3, and 4 weeks were 12.5 percent, 27 percent, 35 percent, and 50 percent, respectively. In the VEGF (165) treated group, survival rates were 14.8 percent, 37 percent, 48 percent, and 74.3 percent, respectively. Statistically significant differences were noted between the two groups at the 2-week ( p = 0.047), 3-week ( p = 0.048), and 4-week ( p = 0.023) time intervals. The authors conclude that the TAIL flap is a novel and useful animal model to study flap prefabrication. PMID- 16254816 TI - Role of simple cold storage in preventing epiphyseal growth plate impairment after replantation surgery in immature rats. AB - The purpose of this study was to investigate the effect of ischemia time on longitudinal bone growth after replantation, and to evaluate the usefulness of simple cold storage of an amputated limb in preventing epiphyseal growth-plate impairment. A hind-limb replantation model was produced with 5-week-old rats after various ischemia times. With more than 6 hr of warm ischemia, growth disturbance was observed, and extensive necrosis was histologically apparent in the central region of the growth plate of the proximal tibia, even at 1 week postoperatively. Destruction of the growth plate was complete at 4 weeks after surgery. By simple cold storage of the amputated limb, growth disturbance was not observed, even following ischemia for 9 hr, and no abnormal findings were observed histologically. Segmental necrosis and destruction of the growth plate observed histologically suggested the possibility of indirect impairment of the epipyseal chondrocytes due to disturbance of the epiphyseal arterial system, attributable to ischemia. The study confirmed experimentally that cold storage of the amputated part may prevent longitudinal bone-growth disturbance after extension of ischemia time. PMID- 16254817 TI - Microsurgical training model for nerve repair. PMID- 16254818 TI - Repeated oral once daily intake of increasing doses of the novel synthetic genistein product Bonistein in healthy volunteers. AB - Bonistein is a new product consisting of > 99.5 % synthetic genistein, an isoflavone with phyto-oestrogenic properties, which might be a safe and efficacious alternative for the prevention of post-menopausal bone loss to the traditional hormone replacement therapy. A randomised, open-labelled and sequential-group phase I study was performed to assess safety, tolerability and pharmacokinetic characteristics of oral administrations of Bonistein. Thirty healthy volunteers received in three subsequent groups 30, 60 or 120 mg once daily for 14 days. For the pharmacokinetic profiles of Bonistein, blood samples were taken on study Days 1 (after first dose) and 14 (steady state). Repeated intake of Bonistein was well tolerated. A total of 33 adverse events were reported, mainly of mild intensity. No relevant changes in clinical laboratory or vital signs were observed. The pharmacokinetic characteristics of Bonistein revealed comparable results for extent and rate of absorption on Days 1 and 14. Both AUC and C (max) values of Bonistein increased in proportion with the dose. PMID- 16254819 TI - 8-NH2-boldine, an antagonist of alpha1A and alpha1B adrenoceptors without affinity for the alpha1D subtype: structural requirements for aporphines at alpha1-adrenoceptor subtypes. AB - Structure-activity analysis of 21 aporphine derivatives was performed by examining their affinities for cloned human alpha (1A), alpha (1B) and alpha (1D) adrenoceptors (AR) using membranes prepared from rat-1 fibroblasts stably expressing each alpha (1)-AR subtype. All the compounds tested competed for [ (125)I]-HEAT binding with steep and monophasic curves. The most interesting compound was 8-NH (2)-boldine, which retains the selective affinity for alpha(1A) AR (pKi = 6.37 +/- 0.21) vs. alpha(1B)-AR (pKi = 5.53 +/- 0.11) exhibited by 1,2,9,10-tetraoxygenated aporphines, but shows low affinity for alpha(1D)-AR (pKi < 2.5). Binding studies on native adrenoceptors present in rat cerebral cortex confirms the results obtained for human cloned alpha (1)-AR subtypes. The compounds selective for the alpha (1A) subtype discriminate two binding sites in rat cerebral cortex confirming a mixed population of alpha (1A)- and alpha (1B) AR in this tissue. All compounds are more selective as inhibitors of [ (3)H] prazosin binding than of [ (3)H]-diltiazem binding to rat cerebral cortical membranes. A close relationship was found between affinities obtained for cloned alpha (1A)-AR and inhibitory potencies on noradrenaline-induced contraction or inositol phosphate accumulation in tail artery, confirming that there is a homogeneous functional population of alpha(1A)-AR in this vessel. On the contrary, a poor correlation seems to exist between the affinity of 8-NH (2) boldine for cloned alpha (1D)-AR and its potency as an inhibitor of noradrenaline induced contraction or inositol phosphate accumulation in rat aorta, which confirms that a heterogeneous population of alpha (1)-AR mediates the adrenergic response in this vessel. PMID- 16254820 TI - An anti-inflammatory ent-kaurane from the stems of Annona squamosa that inhibits various human neutrophil functions. AB - We have previously shown that 11 ent-kauranes isolated from the stems of Annona squamosa exhibited immunomodulating effects in leukocytes. In this study, a cellular model using isolated human neutrophils, which are important in the pathogenesis of rheumatoid arthritis, ischemia-reperfusion injury, chronic obstructive pulmonary disease, asthma and other inflammatory diseases, was established in order to elucidate the anti-inflammatory functions of 16beta,17 dihydroxy-ent-kauran-19-oic acid (1). Reactive oxygen species (ROS) and granule proteases produced by neutrophils contribute to the pathogenesis of inflammatory diseases. Compound 1 inhibited the generation of superoxide anion, the formation of ROS, and the release of elastase in formyl-L-methionyl-L-leucyl-L phenylalanine (FMLP)-activated human neutrophils in a concentration-dependent manner with IC (50) values of 3.95 +/- 0.68, 12.20 +/- 2.16, and 12.52 +/- 2.26 microM, respectively. The anti-inflammatory actions were not attributable to cytotoxicity because incubation of the neutrophils with 1 did not result in lactate dehydrogenase release. Compound 1 did not display antioxidant or superoxide anion-scavenging activity. Furthermore, neither subcellular NADPH oxidase activity nor cAMP-dependent pathways were altered by 1. Compound 1 significantly inhibited rapid calcium release from internal calcium stores induced by FMLP but not by thapsigargin. In summary, the presented results indicate that the inhibitory effects of 1 on respiratory burst and degranulation of human neutrophils are through the inhibition of cytosolic calcium mobilization, but not via the cAMP-dependent pathways. PMID- 16254821 TI - A Vitex agnus-castus extract inhibits cell growth and induces apoptosis in prostate epithelial cell lines. AB - Extracts of Vitex agnus-castus fruits (VACF) are described to have beneficial effects on disorders related to hyperprolactinemia (cycle disorders, premenstrual syndrome). A VACF extract has recently been shown to exhibit antitumor activities in different human cancer cell lines. In the present study, we explored the antiproliferative effects of a VACF extract with a particular focus on apoptosis inducing and potential cytotoxic effects. Three different human prostate epithelial cell lines (BPH-1, LNCaP, PC-3) representing different disease stages and androgen responsiveness were chosen. The action of VACF on cell viability was assessed using the WST-8-tetrazolium assay. Cell proliferation in cells receiving VACF alone or in combination with a pan-caspase inhibitor (Z-VAD-fmk) was quantified using a Crystal Violet assay. Flow cytometric cell cycle analysis and measurement of DNA fragmentation using an ELISA method were used for studying the induction of apoptosis. Lactate dehydrogenase (LDH) activity was determined as a marker of cytotoxicity. The extract inhibited proliferation of all three cell lines in a concentration-dependent manner with IC (50) values below 10 microg/mL after treatment for 48 h. Cell cycle analysis and DNA fragmentation assays suggest that part of the cells were undergoing apoptosis. The VACF-induced decrease in cell number was partially inhibited by Z-VAD-fmk, indicating a caspase-dependent apoptotic cell death. However, the concentration-dependent LDH activity of VACF treated cells indicated cytotoxic effects as well. These data suggest that VACF contains components that inhibit proliferation and induce apoptosis in human prostate epithelial cell lines. The extract may be useful for the prevention and/or treatment not only of benign prostatic hyperplasia but also of human prostate cancer. PMID- 16254822 TI - Crocetin prevents dexamethasone-induced insulin resistance in rats. AB - The main objective of the study was to examine whether crocetin, a natural product from Gardenia jaminoides Ellis, has beneficial effects on the state of insulin resistance induced by dexamethasone in a rat model. Measured using the oral glucose tolerance tests (OGTT), male Wistar rats treated with subcutaneous dexamethasone (0.08 mg/kg/d) for 6 weeks exhibited reduced insulin sensitivity at weeks 2 and 4 and impaired glucose tolerance at week 4. In the dexamethasone treated group, serum insulin, free fatty acids (FFA), triglyceride (TG) and tumor necrosis factor (TNF)-alpha levels were significantly increased at the end of the study. In addition, the hepatic glycogen content was reduced as indicated by periodic acid-Schiff (PAS) staining, and pancreatic islet beta cells showed compensatory hyperactivity suggested by immunohistochemical (IHC) staining using an antibody against insulin. Treatment with crocetin (40 mg/kg/d) significantly attenuated all the described effects of dexamethasone. These results suggest that crocetin might prevent the development of dexamethasone-induced insulin resistance and related abnormalities in rats. PMID- 16254823 TI - Cytotoxic and genotoxic butanolides and lignans from Aiouea trinervis. AB - The ethanolic extracts from the roots, the underground trunk and the leaves of Aiouea trinervis were active in the brine shrimp (Artemia salina) lethality assay (LD (50): 1.93, 0.92 and 262.1 microg/mL, respectively). Fractionation of the extracts led to the isolation of four butanolides, namely (-)-epilitsenolides C (2) and C (1) ( 1 and 2), isoobtusilactone A ( 3) and obtusilactone A ( 4), two of which ( 1 and 2) are reported for the first time as genuine natural products. The lignans (+)-sesamin ( 5) and (+)-methylpiperitol ( 6) and polyprenol-12 ( 7) were isolated as well. Their structures were determined with spectral methods (1D , 2D-NMR and MS). Compounds 1, 2, 3, 5 and 6 were tested for their cytotoxic activities in Hep (2) human cancer cells. The butanolides 2 and 3 were the most active (IC (50): 5.96 microg/mL and 4.95 microg/mL, respectively) whereas the other compounds showed moderate IC (50) values ranging from 12.20 microg/mL to 25.64 microg/mL. The genotoxic properties of the crude ethanolic extracts and of compounds 3 and 5 were also evaluated in this study on CHO K1 and HTC mammalian cells with single-cell gel electrophoresis (comet assay). The crude extracts as well as the compounds tested induced DNA migration in this assay, which was indicative of DNA damage (genotoxic effect). PMID- 16254824 TI - Isolation of new rotenoids from Boerhaavia diffusa and evaluation of their effect on intestinal motility. AB - A bioassay-guided separation of a methanolic extract obtained from the roots of Boerhaavia diffusa L. (Nyctaginaceae) allowed us to isolate five compounds belonging to the class of rotenoids: the known boeravinone D ( 1), boeravinone E ( 2), compound 5 and two novel compounds that we have named boeravinone G ( 3) and boeravinone H ( 4). The structures of the new molecules have been determined on the basis of their HR-EI-MS, (1)H- and (13)C-NMR and 2D-NMR (HMQC, HMBC) data. All the isolated rotenoids have been evaluated for their effect on intestinal motility in vitro. Three of them (boeravinone G, boeravinone E and compound 5) exhibited spasmolytic activity. Preliminary structure-activity relationships have been established highlighting the effect of substitutions on rings B and D. PMID- 16254825 TI - Effect of three flavonoids isolated from Japanese Polygonum species on superoxide generation in human neutrophils. AB - Three flavonoids, quercetin 3- O-glucoside-2''-gallate (QGG), quercetin 3- O rhamnoside-2''-gallate (QRG) and kaempferol 3- O-glucoside-2''-gallate (KGG) were isolated from Japanese Polygonum species. The effect of these flavonoids on stimulus-induced superoxide generation in human neutrophils was assayed by measuring the reduction of cytochrome c. The tyrosyl or serine/threonine phosphorylation of neutrophil proteins and the translocation of p4(phox) and p67(phox) to the cell membrane were detected using specific monoclonal antibodies. The flavonoids used in this experiment significantly suppressed stimulus-induced superoxide generation in a concentration-dependent manner. FMLP induced tyrosyl phosphorylation or PMA-induced serine/threonine phosphorylation and the translocation of the cytosolic proteins p47(phox) and p67(phox) to the cell membrane were suppressed in parallel to the suppression of the stimulus induced superoxide generation. PMID- 16254826 TI - Modulation of the G2 cell cycle checkpoint by sesquiterpene lactones psilostachyins A and C isolated from the common ragweed Ambrosia artemisiifolia. AB - A phenotypic cell-based assay for inhibitors of the G (2) DNA damage checkpoint was used to screen plant extracts from the US National Cancer Institute Natural Products Repository. It revealed activity in a methanol extract from the common ragweed Ambrosia artemisiifolia. Assay-guided fractionation led to the identification of the sesquiterpene lactones psilostachyins A and C as novel checkpoint inhibitors. Elimination of their alpha,beta-unsaturated carbonyl group caused a loss of activity, suggesting that the compounds can bind covalently to target proteins through Michael addition. Psilostachyins A and C also blocked cells in mitosis and caused the formation of aberrant microtubule spindles. However, the compounds did not interfere with microtubule polymerization in vitro. The related sesquiterpene lactones psilostachyin B, paulitin and isopaulitin were also isolated from the same extract but showed no checkpoint inhibition. The identification of the target(s) of psilostachyins A and C may provide further insight into the signalling pathways involved in cell cycle arrest and mitotic progression. PMID- 16254827 TI - Embelin derivatives and their anticancer activity through microtubule disassembly. AB - Biological activities of the 1,4-benzoquinone derivatives 5- O-ethylembelin ( 1) and 5- O-methylembelin ( 2) were investigated. Both of them showed antiproliferative activity against a panel of human tumor cell lines upon comparison to normal marsupial kidney cells (PtK2). They arrested HL-60 cells in the G(0)/G(1) phase of the cell cycle in a dose- and time-dependent manner. In HeLa cells, exposure to 100 microM of 1 or 2 for 6 h induced a complete disassembly of the microtubule network and an increased number of cells blocked in mitotic stages. Treatment with 10 microM of 1 and 2 for 24 h induced apoptosis in HL-60 cells. This evidence suggests that both 1 and 2 are promising novel antimitotic and anticancer molecules targeting microtubular proteins. PMID- 16254828 TI - Sesquiterpene lactones with potent cytotoxic activities from Vernonia chinensis. AB - Five new sesquiterpene lactones, namely vernchinilides A-E (1-5), along with five known compounds, 8 beta-(2-methylacryloyloxy)hirsutinolide 13-O-acetate (6), 8 alpha-(2-methylacryloyloxy)-1beta,4beta-epoxy-1alpha-methoxy-13-O-acetate-10 betaH-germacra-5 E,7(11)-dien-12,6-olide (7), 8 beta-(2 hydroxymethylacryloyloxy)hirsutinolide 13-O-acetate (8), 8 alpha tigloyloxyhirsutinolide 13-O-acetate ( 9) and vernolide-B (10) were isolated from Vernonia chinensis. The structures of these new compounds were elucidated on the basis of spectral data, especially 2D-NMR techniques. Compounds 2, 5 and 6 exhibited potent cytotoxic activities against P-388 and A-549 tumor cell lines. PMID- 16254829 TI - Cytotoxic phenolic constituents from the root of Actinidia chinensis. AB - Twelve phenolic compounds, including four novel skeleton phenolic compounds, planchols A-D (1-4), together with four pairs of isomeric flavanoids (5-12) were isolated from the root of Actinidia chinensis Planch (Actinidiaceae). Their structures were elucidated by spectroscopic analysis and chemical evidence. The structure of 1 was further confirmed by a single-crystal X-ray diffraction determination. Moreover, it was found that 1 and 2 showed remarkable cytotoxic activity against P-388 and A-549 cell lines. PMID- 16254830 TI - New acylated clionasterol glycosides from Valeriana officinalis. AB - The chloroform extract of Valeriana officinalis led to the isolation of clionasterol-3-O-beta-D-glucopyranoside and a mixture of 6'-O-acyl-beta-D glucosyl-clionasterols. The acyl moieties were identified as hexadecanoyl, 8 E,11 E-octadecadienoyl and 14-methylpentadecanoyl by alkaline hydrolysis followed by GC-MS analysis. The isolated compounds did not exhibit any anti-inflammatory, anticancer or cytotoxic activity when tested in a variety of in vitro cell-based assays. PMID- 16254831 TI - The inhibitory effect of the flavonoid galangin on urinary bladder smooth muscle contractility is mediated in part by modulation of Ca2+ release from intracellular stores. AB - The present study was designed to examine the effect of the flavonoid galangin on the muscarinic receptor mediating a carbachol-induced contraction and to investigate the effect of the flavonoid on Ca (2+) release from intracellular stores in the urinary bladder of the pig. Galangin (10(-7) -10(-4)M) produced a concentration-dependent inhibition of the contractile responses to electrical field stimulation (EFS) and carbachol (10(-5)M). Galangin (3 x 10(-5)M) reduced muscle contractions evoked by carbachol (10(-5)M) in calcium-containing solution as well as contractions evoked by carbachol and caffeine (2 x 10(-2)M) in Ca(2+) free solutions significantly. The flavonoid had a stronger effect on the maximal force of the contractions induced by caffeine, compared to contractions induced by carbachol. These results suggest that galangin has an important effect on the intracellular calcium mobilization, which might be attributed predominantly to its influence on ryanodine-receptors. PMID- 16254833 TI - Semisynthetic 15-O-acyl- and 1,15-di-O-acyleurycomanones from Eurycoma longifolia as potential antimalarials. AB - Among the quassinoids isolated from Eurycoma longifolia Jack, eurycomanone was identified as the most potent and toxic inhibitor of the chloroquine-resistant Gombak A isolate of Plasmodium falciparum. Several diacylated derivatives of eurycomanone, 1,15-di-O-isovaleryleurycomanone, 1,15-di-O-(3,3-dimethylacryloyl)- eurycomanone and 1,15-di-O-benzoyleurycomanone were synthesized by direct acylation with the respective acid chlorides. The monoacylated 15-O isovaleryleurycomanone was synthesized by selective protection of the other hydroxy groups of eurycomanone with trimethylsilyl trifluoromethanesulphonate to enable the exclusive acylation of its C-15 hydroxy group. This was followed by the removal of the protecting groups with citric acid. The diacylated eurycomanones exhibited lower antiplasmodial activity against the Gombak A isolates and lower toxicity in the brine shrimp assay when compared to eurycomanone. In contrast, the monoacylated derivative displayed comparable antiplasmodial potency to eurycomanone, but its toxicity was reduced. Thus, preliminary studies of the synthesized acylated eurycomanones have shown that acylation only at the C-15 hydroxy group may be worthy of further antimalarial investigation. PMID- 16254832 TI - Potential antimalarial lead structures from fungi of marine origin. AB - Antiplasmodial and cytotoxicity testing of five highly oxygenated natural products (6R,12R,14R-colletoketol, 6R,11R,12R,14R-colletoketodiol, dihydrobotrydial, pycnidione, and 3R,4S-hydroxymellein), all derived from fungi of marine origin, showed one of them, pycnidione, to have activities against three different strains of Plasmodium falciparum in the sub-micromolar (microM) range. Although the mean selectivity index of 1 for the observed antiplasmodial activity of 4 is low, pycnidione's usefulness as a potential lead structure should not be ignored. PMID- 16254834 TI - Cytotoxic chalcones and flavonoids from the leaves of Muntingia calabura. AB - Two new dihydrochalcones, 2',4'-dihydroxy-3'-methoxydihydrochalcone, (-)-3' methoxy-2',4',beta-trihydroxydihydrochalcone, a new flavanone, (2 S)-(-)-5' hydroxy-7,3',4'-trimethoxyflavanone, and a new flavonol derivative, muntingone, along with sixteen known compounds, were isolated from the leaves of Muntingia calabura. The structures of these new compounds were determined using spectral analyses including extensive 2D NMR data. Among the isolates, (2 S)-5'-hydroxy 7,3',4'-trimethoxyflavanone, 4'-hydroxy-7-methoxyflavanone, 2',4' dihydroxychalcone, and 2',4'-dihydroxy-3'-methoxychalcone exhibited cytotoxicity (IC (50) values < 4 microg/mL) against P-388 and/or HT-29 cell lines in vitro. PMID- 16254835 TI - Two new antioxidant stilbene dimers, parthenostilbenins A and B from Parthenocissus tricuspidata. AB - The ethyl acetate fraction of an aqueous alcoholic extract from the stem of Parthenocissus tricuspidata yielded 11 known compounds (1-11) and two new stilbene dimers parthenostilbenins A (12) and B (13) upon purification either by preparative TLC or reversed phase HPLC. The structures of the new isolates were identified using a combination of FAB-MS and NMR. These compounds were assessed for antioxidant activities in three different bioassay systems. Among them, piceatannol showed the strongest inhibitory activity in these assay systems. Two new compounds, parthenostilbenins A (12) and B (13) inhibited lipid peroxidation (IC (50) = 20.35 +/- 1.22 and 18.68 +/- 0.51 microg/mL, respectively) in a rat liver homogenate. PMID- 16254836 TI - Phenolic glycosides from Potalia amara. AB - Investigation of the stem bark of the unique Amazonian herbal plant Potalia amara yielded two new phenolic glycosides, potalioside A (1) and B (2), along with di-O methylcrenatin (3), 2,6-dimethoxy-4-hydroxyphenol 1-glucoside and sweroside. The structures of potalioside A and B were established by interpretation of spectral data as 4-hydroxymethyl-2,6-dimethoxyphenyl 1-O-beta-D-glucopyranosyl(1-->6)-beta D-glucopyranoside and 4-hydroxymethyl-2,6-dimethoxyphenyl 1-O-beta- D xylopyranosyl(1-->6)- beta-D-glucopyranoside, respectively. PMID- 16254837 TI - Acetophenone derivatives and sesquiterpene from Euphorbia ebracteolata. AB - Two new acetophenone derivatives and a new sesquiterpene, along with four known acetophenone compounds, were isolated from the roots of Euphorbia ebracteolata. The structures of the new compounds were identified as 2,2',4,4'-tetrahydroxy 6,6'-dimethoxy-3,3'-dimethyl-7,5'-bisacetophenone, 2,4-dihydroxy-6-methoxy-3 methylacetophenone 4-O-alpha-L-arabinofuranosyl-(1-->6)-beta- D-glucopyranoside and decahydro-1a,4a,7,7-tetramethyl-1H-cycloprop[e]azulene-2,5-diol (1a R,2 R,4a S,5 S,7a R,7b R) using 1D and 2D NMR spectroscopy and X-ray crystallography. PMID- 16254838 TI - Four new cucurbitane glycosides from Hemsleya jinfushanensis. AB - A phytochemical study of the tubers of Hemsleya jinfushanensis L. T. Shen resulted in the isolation of four new cucurbitane glycosides, jinfushanosides A-D (1- 4), as well as four known compounds 5-8. Compounds 1-7 were tested for bioactivity against rabbit platelet aggregation induced by PAF, ADP, or AA. Among them, compounds 1, 5, 6 and 7 weakly inhibited PAF-induced platelet aggregation. PMID- 16254839 TI - Molecular cloning and expression of putrescine N-methyltransferase from the hairy roots of Anisodus tanguticus. AB - A full-length cDNA encoding putrescine N-methyltransferase (PMT) was isolated from the hairy roots of A. tanguticus. Nucleotide sequence analysis of the cloned cDNA revealed an open reading frame of 1017 bp encoding 338 amino acids with high homology to other known PMTs. A. tanguticus PMT was expressed in Escherichia coli. The recombinant AtPMT was purified and exhibited S-adenosyl-methionine dependent N-methyltransferase activity. PMID- 16254841 TI - Minimally invasive urologic surgery in children: an overview of what can be done. AB - PURPOSE: Laparoscopy has emerged as a feasible and effective alternative for abdominal pathologies in children. Urological minimally invasive surgery is now commonly used for basic operations in most centres and is just beginning to expand its use in more complex operations in several selected centres around the world. We present the current state of the art of minimally invasive surgery in children. MATERIAL AND METHODS: We first reviewed all the urological pathologies treated with minimally invasive surgery in our institution. Secondly we reviewed the literature in order to highlight and discuss certain relevant articles and compare them with our own experience. RESULTS: Since beginning to use minimally invasive surgery at our institution we have operated more than 550 cases with urological pathologies. We used three approaches: the transperitoneal, the retroperitoneal and the transvesical approach. Our preferred indications for each approach are discussed. DISCUSSION: Non palpable testis, varicocele surgery, nephrectomy and adrenalectomy are, in our opinion, established minimally invasive procedures. Hemi-nephrectomy, pyeloplasty and Cohen antireflux surgery are probably excellent indications when minimally invasive surgery is carried out by expert hands. Stone management should not be forgotten in cases of contraindication or failure of ESWL. Even the most complex urological operations may be safely carried out using a minimally invasive approach, although most of the cases described are case reports. CONCLUSION: More papers are published on ablative or reconstructive urological minimally invasive surgery. Transperitoneal and retroperitoneal approaches are used with the same results. Transvesicoscopic surgery should rapidly grow to become a standard approach for Cohen reimplantation. It is anticipated that technical progress will provide the opportunity for more paediatric urologists to develop a minimally invasive approach. PMID- 16254842 TI - Thoracoscopic surgery in infants and children. AB - Numerous investigators have shown that video-assisted thoracoscopic surgery (VATS) can be safely used for specific conditions of newborns, infants, and children. The technique has been postulated to be associated with a lower morbidity, shorter hospital stay, lower costs, and clinical results similar to those achieved by open surgery. The present article reviews the state of the art of VATS for thoracic conditions in children. Most authors focus on the feasibility of single procedures, and only a small number of reports deals with the feasibility in series with multiple types of procedures and larger numbers of patients. Therefore, systematic research on the advantages and limits of VATS in children remains mandatory. PMID- 16254843 TI - Laparoscopic surgery in infants and children. AB - Almost all operations that are classically performed as open surgery have now an endoscopic surgical variant. The reason for performing this form of surgery obeys the Hippocratic principle: The less invasive the better. Moreover there is scientific evidence that the less trauma, the less stress response and the less immunosuppression. There are few well conducted studies in children comparing open with endoscopic surgery, but evidence is piling up, especially from studies in adults, that endoscopic surgery results in a faster recovery, better cosmesis and fewer adhesions. The complication rate seems, however, slightly higher. Endoscopic surgery takes more time but hospital stay is shorter so that the same output can be achieved with fewer beds. Endoscopic surgery has changed the mentality of pediatric surgeons. Nowadays pediatric surgeons think more in terms of invasiveness which means that even in open surgery incisions are not as large anymore as they have been in the past. Endoscopic surgery has also changed the mentality in nursing. The wounds do not anymore reflect the magnitude of the surgery that has been performed internally, which may lead to an underestimation of pain. As the turnover of patients is much higher, there is less patient and parent binding. Lastly some parents may be overwhelmed by the rapid discharge and nurses have to guide them. Some nurses feel that the ward has become less surgical, which may have an impact on recruitment. PMID- 16254844 TI - Effects of whole-body hypoxic preconditioning on hypoxia/reoxygenation-induced intestinal injury in newborn rats. AB - PURPOSE: The precise cause of necrotizing enterocolitis (NEC) is elusive. Ischemia and reperfusion injury of the intestine has been considered to be a major contributing factor for NEC. Ischemic preconditioning is defined as one or more brief periods of ischemia with intermittent reperfusion that protects tissues against a sustained period of subsequent ischemia. Contribution of preconditioning to hypoxia/reoxygenation-induced intestinal injury in newborn rats has not been evaluated previously. METHODS: The study was carried out on 1 day-old Wistar albino rat pups. Whole-body hypoxia and reoxygenation (H/R) was achieved by 10 min hypoxia using 95 % N (2) + 5 % CO (2) followed by 10 min reoxygenation with 100 % oxygen. Whole body hypoxic preconditioning (HP) cycles were performed with 3 min hypoxia and 5 min reoxygenation. Thirty-three pups were randomly allocated into 4 groups. Group 1 served as untreated controls. The pups in group 2 were subjected to H/R only. In groups 3 and 4, 1 cycle and 3 cycles of HP were performed prior to H/R, respectively. Animals were killed at the end of the protocols. Intestine specimens were obtained to determine the histological changes, as well as to measure the tissue malondialdehyde (MDA) and nitric oxide (NO) levels, and xanthine oxidase (XO) and myeloperoxidase (MPO) activities. RESULTS: The microscopic lesions in H/R rat pups were virtually the same as those seen in neonatal NEC, with severe destruction of villi and crypts, in some cases extending to the muscularis. In both HP groups, the lesions were found to be milder. H/R resulted in a marked elevation in MDA and NO levels, and XO and MPO activities compared to the untreated controls. Both 1 cycle and 3 cycles of HP prior to H/R resulted in an obvious decrease in all biochemical parameters. Differences of the biochemical results between both HP groups were not statistically significant. CONCLUSION: This study revealed that whole-body hypoxic preconditioning is beneficial for hypoxia/reoxygenation-induced intestinal injury in newborn rats. PMID- 16254845 TI - Resveratrol may reduce apoptosis of rat testicular germ cells after experimental testicular torsion. AB - The aim of the study is to evaluate the effects of resveratrol on apoptosis of testicular germ cells after experimental testicular torsion. Thirty Wistar albino rats were divided into 3 groups. Torsions were created by rotating the right testis 720 degrees in a clockwise direction for 4 h in all groups except the control group (group 1). They were then repaired by counter-rotation and replaced into the scrotum. In group 2, saline was infused 30 min before detorsion. In group 3, 30 mg/kg resveratrol was infused 30 min before detorsion. In groups 2 and 3, the bilateral testes were removed to determine germ cell apoptosis after 20 h of detorsion. The number of apoptotic cells was evaluated using the terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling (TUNEL) technique and caspase 3. Mean apoptotic score of ipsilateral testes in group 3 was lower than that of group 2 (p < 0.05). Mean apoptotic score of the contralateral testes in group 3 was not different from that of group 2 (p > 0.05). The present study demonstrates that intraperitoneal administration of resveratrol in rats may decrease germ cell apoptosis in the ipsilateral testes. PMID- 16254846 TI - Lymphatic regeneration following hind limb replantation: an experimental study in the dog. AB - Different types of trauma to the lymphatic system can often occur, but surgical intervention can be performed only in specific cases. We report on lymphatic regeneration following limb replantation in traumatic amputations and replantation of extremities. The aim of this study was to observe the progression and reaction after surgical trauma that is similar to other kinds of trauma, both in children or adults, and to monitor the possible lymphatic regeneration. Particular attention was paid to two parameters: firstly, the physical examination of the replanted limbs by checking the post-traumatic lymphoedema, and secondly, the study of the images taken from indirect lymphangiography of the replanted limbs. Histological specimens of the surgical trauma area were also examined to reconfirm or exclude lymphatic regeneration. The study population consisted of sixteen mongrel dogs, divided into two groups of eight animals each, who underwent hind limb elective amputation and replantation combined with (group A) or without (group B) sciatic nerve division. Lymphoedema formation was followed quantitatively by measurement of the circumference of the hind limb for 21 days after replantation. Indirect lymphography, never performed before in such cases, and histopathology, were performed to evaluate and confirm lymphatic regeneration. Lymphatic regeneration after replantation of the operated hind limbs was first confirmed between 7th and 11th postoperative day by indirect lymphangiography and clinical observation of the post-traumatic lymphoedema of these limbs. The mean time of visualisation of lymphatic regeneration through lymphography was 10.12 days for group A and 9.37 days for group B. However, nerve transection had no effect on lymphatic regeneration (p = 0.46). Histopathological examination showed first evidence of lymphatic regeneration on the ninth postoperative day and a network of newly formed capillary lymphatics on the 21st postoperative day. It is concluded that lymphatic regeneration following replantation of the extremities without anastomosing of the interrupted lymph vessels, is an unquestionable fact. To achieve the best lymphatic drainage and use of the replanted extremities it is important to resect all non-vital tissues of the replantation area. Local or general infections decelerate lymphatic regeneration. Indirect lymphography with iotrolan is a reliable, easy to perform technique without complications that may be used repeatedly for confirmation and evaluation of post-traumatic lymphoedema. PMID- 16254847 TI - Functional imaging for determination of nephroblastoma: an obligatory diagnostic program. AB - In Europe, 1.7 % of children with predicted nephroblastoma undergo neoadjuvant chemotherapy without being correctly diagnosed. It is necessary to reduce the number of misdiagnoses. In a girl who was referred to our institute with the diagnosis of nephroblastoma of the right kidney, DMSA scan revealed a hypertrophic compensation of the contralateral nephropathy in the lower part. Functional imaging studies of the kidney should be performed in patients in whom the type of pathology of a suspect mass in the kidney region is unclear, including a careful scrutiny of the MRI scan. PMID- 16254848 TI - Urodynamics-based evidence for the beneficial effect of imipramine on valve bladders in children. AB - AIMS: In an era, in which valve bladder is recognised as perhaps the single most important determinant of long-term outcome in patients with posterior urethral valves (PUV), an insight into the etiopathogenesis and management of valve bladders is warranted. The present study was designed to evaluate bladder dysfunction in PUV and to assess the response to imipramine in these patients, both subjectively and objectively, by serial urodynamic studies (UDS). METHODS: From 1998-2001, 30 patients with PUV who had documented bladder dysfunction on UDS were studied. Patients with non-compliant or unstable bladders were treated with imipramine (1.5 - 2 mg/kg). All the patients in the present study were 5 years or older and hence old enough to be toilet trained. Assessment of continence and side effects of the drug was done after 3 months and repeat UDS were done at 3-6 months, 1 and 2 years. RESULTS: On the basis of initial treatment, the patients were divided into 2 groups; a fulguration group (n = 10, 33.3 %) and a vesicostomy group (n = 20, 66.6 %). Symptomatic voiding dysfunction was present in 27 of the 30 patients (90 %). Two patterns of urodynamic abnormalities were noted in the present study; 1) unstable bladders with single or multiple uninhibited contractions (18/30 patients, 60 %), and 2) small capacity, hypocompliant, hypertonic bladder (12/30 patients, 40 %). Post imipramine therapy significant symptomatic improvement was noted in 16/30 patients. On serial UDS, there was a 18-20 % increase in maximum cystometric capacity (MCC) and 30-35 % increase in pressure specific bladder volume (PSBV) following one year of imipramine therapy in 16/30 patients and 11/30 patients, respectively. 4 patients failed to show any improvement in MCC and PSBV with imipramine, they had been initially diverted with vesicostomy and later required augmentation cystoplasty. CONCLUSION: Unstable bladders and those with marginal bladder capacity and compliance showed the best response to imipramine therapy. Fibrotic, small capacity, hypertonic bladders are less responsive to imipramine. However, a trial of imipramine therapy is still warranted in these patients, as only 4/12 (33.3 %) patients with fibrotic hypertonic bladders failed to show any response and ultimately required augmentation cystoplasty. Imipramine qualifies as an effective and cheap drug for valve bladders. PMID- 16254849 TI - Esophageal atresia with double tracheoesophageal fistula--a case report and review of the literature. AB - Esophageal atresia with double tracheoesophageal fistula (TEF) is a very rare anomaly, and the accurate preoperative diagnosis of proximal TEF is very difficult. This paper describes a baby girl who presented with esophageal atresia with double, proximal, and distal TEF. The distal TEF was diagnosed before operation, whereas the proximal TEF was found intraoperatively. Overlooking the presence of proximal TEF can lead to increased morbidity and mortality due to severe respiratory infection and the necessity of a second operation. Great care must therefore be taken to not overlook the presence of proximal TEF in patients with this anomaly. PMID- 16254850 TI - Acute portal vein thrombosis after splenic embolization and splenectomy for autoimmune pancytopenia. AB - Postsplenectomy portal vein thrombosis for hematological diseases is uncommon in the pediatric population. The case summarized is, to our knowledge, the first manifestation of portal vein thrombosis in a child after preoperative splenic artery embolization and subsequent splenectomy for severe hypersplenism. We suggest that early routine diagnosis by Doppler ultrasonography and subcutaneous low molecular weight heparin therapy are useful steps for a successful outcome. PMID- 16254851 TI - Spontaneous rupture of extrahepatic choledochal cyst: two pediatric cases and literature review. AB - Spontaneous rupture of extrahepatic choledochal cysts is very rare. Neonatal cases generally present with biliary ascites, and older children with acute abdomen. Although the cause is unclear, accumulation of protein plugs in an anomalous pancreaticobiliary junction, irritation of the cyst wall due to reflux of pancreatic juice, and weakness due to a developmental error such as common channel syndrome have all been suggested to play contributing roles. The treatment of choice is complete excision of the cyst and hepatico-jejunostomy. In newborns who are in a poor condition, the primary treatment should be simple peritoneal drainage. PMID- 16254852 TI - Annular pancreas in two consecutive siblings: an extremely rare case. AB - Annular pancreas is the rare congenital anomaly where the pancreas forms a full or incomplete ring around the second segment of the duodenum, causing various degrees of stenosis or atresia. It is estimated that it appears in 1 out of 12 000-15 000 births of living neonates and until now, in the literature, only 6 cases have been reported among individuals of the same family. We present the case of two siblings, a boy and a girl, with annular pancreas from consecutive pregnancies of the same couple. Both neonates had a prenatal diagnosis of duodenal obstruction and they underwent duodenoduodenal, proximal transverse to distal longitudinal anastomosis. Furthermore, the girl had a mobile ascending colon. Their postoperative condition was perfect. The case we are reporting is an addition to the other 6 cases of familial presentation of annular pancreas and is similar to one of them. In these families, a total of 16 persons present this congenital anomaly while 14 are seemingly healthy. Twelve of the affected persons are female and 4 male. In conclusion, it can be stated that female individuals seem to have a greater propensity to transmit the disease to their descendants, compared to males, suggesting the possible action of an autosomal recessive sex influenced gene. The recording of such rare family cases should be encouraged, in order to fully recognize a possible type of inherited transmission. PMID- 16254853 TI - Complete Currarino triad in all affected members of the same family. AB - Currarino triad is a rare hereditary disease. The complete form is characterized by the presence of an anorectal malformation, usually anorectal stenosis, a presacral mass and sacral bony defect. The main symptom is the presence of constipation since early life. In over 80 % of cases, the syndrome is diagnosed during the first decade of life. We report on three patients, members of the same family, with the complete form of the syndrome. The main symptom in our patients was intense constipation and the common clinical finding was an ectopic and stenotic anus. In two of them, father and son, the presacral masses were not diagnosed at the time of previous unsuccessful operations in another hospital for correction of the ectopic anus. In the third patient, Currarino syndrome was associated with Hirschsprung's disease. This has not been described previously. Two of the three patients who were admitted in our institution for simultaneous excision of the presacral mass and correction of the anorectal malformation through a posterior midsagittal coccygo-perineal approach, are free of symptoms. To the authors' knowledge, this is the second report of complete Currarino triad in all affected members of the same family. PMID- 16254854 TI - The surgeon in training--European qualifications. PMID- 16254856 TI - [A study of the risk factors in transient global amnesia and its differentiation from a transient ischemic attack]. AB - INTRODUCTION: Transient global amnesia (TGA) is a clinical condition that has been described in many studies, but its pathophysiology is not fully understood. In recent years the theory of valvular insufficiency in the jugular vein has been added to the classical hypotheses that link it to migraine, to epilepsy and -the most widely accepted- to transient ischemic attacks (TIA), although the real origin of the condition has still not been determined. PATIENTS AND METHODS: In a retrospective study we compared 131 patients diagnosed with TGA between 1993 and 2004 with 262 patients who were diagnosed as having TIA over the same period. RESULTS: Mean age was 65.94 years in TGA versus 71.11 years in the case of TIA. There was a higher rate of arterial hypertension among the patients with TGA and diabetes mellitus was more frequent among those with TIA (p<0.05 in both cases). Emboligenic heart disease was scarce among patients with TGA. The number of patients with a history of ischaemic heart disease and a history and the development of cerebrovascular diseases was greater among those with TIA than in cases of TGA (p<0.05). The TGA recurrence rate was 12%. The percentage of pathological findings in the CAT brain scan was higher in patients with TIA (p<0.05). There are no significant differences between patients with TGA and TIA as far as treatment on hospital discharge is concerned. CONCLUSIONS: TGA does not seem to be a symptom of an arteriosclerotic pathology nor does it appear to offer a higher risk of heart or cerebrovascular disease and, therefore, antiaggregating therapy would not be indicated in such cases. PMID- 16254857 TI - [The clinical semiology of attention deficit hyperactivity disorder according to age, and the effectiveness of treatments at different ages]. AB - INTRODUCTION: Attention deficit hyperactivity disorder (ADHD) is the most important cause of poor academic performance and is also usually associated with behavioural, emotional and sociability disorders. AIM: To analyse the different clinical features and the response to psychostimulant therapy, according to the age. SUBJECTS AND METHODS: We reviewed the cases of 152 children referred to Neuropaediatrics owing to suspected ADHD over a 6-year period. Diagnosis and comorbidity were investigated by the looking at their case history, by examination and observation, as well as by interviewing the parents (DSM-IV criteria), and by applying Conners' Rating Scale for parents and teachers, in addition to neuropsychological tests. In order to improve data processing, we established four groups according to the patient's age at diagnosis and when treatment was started: G1: 3-5; G2: 6-8; G3: 9-11; and G4: 12-15 years. RESULTS: ADHD was diagnosed in 102 of the 152 children; 53% had the combined type, 26% were of the inattentional type, and 20% were hyperactive/impulsive. Poor school performance increased with age and reached 75% in G4. The same occurred with conduct disorders: at 6 years of age, 23% were found to have oppositional defiant disorder, and from the age of 9 onwards over 60% of them had conduct disorders. Up until 12 years of age they showed some improvement with psychostimulants and later the rate of dropouts from medical clinical controls and from treatment was over 40%. CONCLUSIONS: The older patients are and the later ADHD is attended, the more problems they have. If successful preventive measures are to be implemented, it would be advisable to begin treatment even before children start primary education (5 years old), so as to try and avoid the pedagogical and behavioural repercussions observed in these children at the age of 6. PMID- 16254858 TI - [Long-term evolution of the doses of Botulinum toxin used in laryngeal and cervical dystonias]. AB - INTRODUCTION: Dystonia is a neurological condition characterised by involuntary movements that give rise to abnormal postures. Different strategies have been used in the treatment of focal dystonias, but the most widely accepted at the present time involves the use of botulinum toxin type A (BTA) injections. Yet, despite its widespread use, the ideal dosages for long-term treatment are still not known with precision. AIMS: The purpose of this study is to report on our experience with long-term BTA therapy in laryngeal (LD) and in cervical dystonia (CD). PATIENTS AND METHODS: We reviewed the data concerning the dosages of BTA injected in 10 patients with LD who received treatment in our centre over a period of eight years. We also analysed the data regarding the doses of BTA injected over an eight-year period in 17 patients with CD. The data were analysed using an ANOVA for paired data. RESULTS: No significant differences were found in the highest dosages of BTA needed for the treatment of LD throughout the progression of the disease (p=0.84). These data contrast with those obtained from the analysis of the treatment of CD, which do show a gradual increase in the dose of toxin that is required (p<0.0001). CONCLUSIONS: These findings suggest that the long-term response to treatment is different in the two conditions. PMID- 16254859 TI - [Primary progressive aphasia: its clinical variability: an analysis of 15 cases]. AB - INTRODUCTION: Primary progressive aphasia (PPA) represents a clinical syndrome linked to multiple degenerative diseases. The diagnosis of PPA is made when language is the only area of salient and progressive dysfunction for at least the first two years of the disease. AIM: To evaluate the neuropsychological, neuropsychiatric and language characteristics of the PPA. PATIENTS AND METHODS: 15 patients with PPA underwent language, neuropsychological and neuropsychiatric evaluation, magnetic resonance imaging, computerized tomography and single photon emission computerized tomography. RESULTS: We observed a clear distinction between the oral expression patterns; the patients were classificated by type of aphasia. The most common sign of PPA was a word finding deficit, also known as anomia. Seven aphasia type Broca, four sensorial transcortical aphasia, two aphasia type Wernicke and two anomic aphasia were found in our patients. Depression, apathy, anxiety and irritability were the most prevalent neuropsychiatric sign. CONCLUSIONS: PPA is a language-based syndrome, that include fluent (normal articulation, flow and number of words per utterance) and nonfluent form of aphasia. It has been considered a cognitive term, however, PPA is associated with high prevalence of psychiatric manifestations. More research it will be necessary to evaluate the prognostic value of them. The slow and progressive deterioration of language provides an interesting model to understand the mechanisms and biological bases involved in the linguistic process. PMID- 16254860 TI - [Linear scleroderma en coup de sabre: neurological symptoms, images and review]. AB - INTRODUCTION: 'Sword stroke' linear scleroderma, which is better known as linear scleroderma en coup de sabre (LSCS), is a rare disease with an uncertain causation that is characterised by progressive craniofacial focal atrophy and is, at least in part, different from Parry-Romberg syndrome (PRS). CASE REPORTS: Here, we report on the cases of 3 patients with LSCS (2 females and 1 male, with a mean age of 40 years). The main neurological symptoms were headache and seizures. Although different alterations were observed in the X-ray images, they were all ipsilateral to the coup de sabre. Histopathological evidence for gliosis and mixed perivascular inflammatory infiltrate was found in the study of a biopsy specimen taken from one female. Cerebrovascular involvement was seen in another patient, as highlighted by the observation of an earlier subclinical cerebellar infarct and occlusion of the superior cerebellar artery in the absence of any other possible causation. CONCLUSIONS: When it affects the central nervous system, the clinical and radiological presentation of LSCS is heterogeneous. Both the imaging studies carried out during the clinical control and the histopathological findings suggest a focal inflammatory process that can be progressive. The arterial involvement is probably due to a non-atherosclerotic, occlusive and chronic inflammatory disease of the peripheral vessels. PMID- 16254861 TI - [The development of the concept of neuronal resting potential. Fundamental and clinical aspects]. AB - INTRODUCTION AND AIMS: Since the classical works carried out on the squid giant axon far less attention has been directed towards the study of the resting membrane potential than to the study of changes in potential (action potential, synaptic potentials, and so forth). It is often assumed that the resting potential depends on an independent current of the voltage called the leakage current, although, except on rare occasions, it has not been possible to characterise such a current either pharmacologically or molecularly. In this work our aim is to review and update the concept of resting potential. DEVELOPMENT: The outlook at present offers a complex situation in which several factors, in addition to leakage currents, play a role in maintaining resting potentials. These factors include ionic currents across non-inactivating voltage-dependent channels, the sodium/potassium pump, and certain currents with characteristics that are similar to those of the classical leakage current. The interaction of all these components gives rise to complex sub-threshold behaviours in the neurons, such as intrinsic rhythmic oscillations, which leads us away from the passive concept of the resting potential. CONCLUSIONS: An ever-increasing number of descriptions of intrinsic sub-threshold, rhythmic activity are being reported, which could suggest that they are a generalised phenomenon in the nervous system. Further studies need to be conducted into the complex mechanisms that determine the resting potential in order to gain an understanding of the phenomena of neuronal excitability and to find an explanation for some of the many pathological conditions that affect the nervous system. PMID- 16254862 TI - [Research into learning disabilities: a documentary analysis]. AB - INTRODUCTION: Our current understanding of learning disabilities (LD) is essentially limited by their actual definition, the heterogeneity of the samples and the unsuitability of the control groups. AIMS: This study offers an overview of the methodological aspects and the areas of research in LD. DEVELOPMENT: From a bibliographical search conducted in the Psychological Abstracts database, PsycINFO, we identified and selected the studies the review was to focus on, and this was completed with previous reviews. Quantitative research methods are preferred in 99.7% of cases, whereas the remaining 0.3% opted for qualitative methods. There was in increase in the number of research reviews, as well as longitudinal studies and those aimed at determining the effectiveness of the interventions, with the resulting interest in new techniques for data analysis such as meta-analysis, path analysis and latent growth curves. The research topics that aroused most interest were LD-related problems (16.7%), difficulties in learning to read (15.82%), studies focusing on intervention (10.15%) and diagnostics in LD. CONCLUSIONS: The primary objectives of the research appear to be the characterisation of LD and the problems they cause throughout the patient's life cycle, together with their identification and early treatment. Research is also extended towards other, more practical matters, with a special emphasis on the school setting. PMID- 16254863 TI - [Early detection and intervention of hearing impairment in Cuba: outcome after 20 years]. AB - INTRODUCTION: Infant hearing loss is a highly prevalent disorder which untreated can severely disrupt normal brain development. As a result there is a significant delay in language acquisition as well as many cognitive and emotional problems in the child. Over the last decades important advances have occurred in the available technology for early detection and assessment of hearing impairment. Therefore many countries worldwide have become aware of the need for hearing screening programs. The optimal protocols depending on the local conditions of health care as well as the availability of technological and human resources. AIM: To summarize the results obtained over the last 20 years by an ongoing hearing screening protocol. DEVELOPMENT: Data on the coverage program, sensitivity and specificity, age of identification of hearing losses, diagnostic and intervention stages will be summarized and discussed. Also the long terms effects of early detection on the child cognitive and language development are analyzed. Finally, the possible role of a new technique based on the recording of multiple auditory steady state potentials with Cuban equipment (AUDIX system) was evaluated within this context. CONCLUSIONS: The Cuban targeting multiple high risk hearing screening program is a useful alternative to early detection of hearing losses. The average detection age of hearing loss was 10 months during the period of optimal functioning. Cognitive, emotional and linguistic development are improved by early detection and intervention. The multiple auditory steady state responses can provide valuable audiometric information within a screening context. PMID- 16254865 TI - [Posterior reversible leukoencephalopathy syndrome caused by cisplatin: a case report]. PMID- 16254866 TI - [Symptoms of parietal epilepsy probably associated to the use of topiramate in a case of temporal epilepsy]. PMID- 16254867 TI - [Clinical control of multiple sclerosis patients treated with interferon beta in a hospital in Peru]. PMID- 16254868 TI - Altered brain activity in women recovered from bulimic-type eating disorders after a glucose challenge: a pilot study. AB - OBJECTIVE: It is not known whether individuals with bulimic-type eating disorders have a dysregulation of brain pathways that modulate appetite. Taste plays a role in the regulation of appetite and the purpose of the current study was to determine whether bulimic women have alterations in the physiologic response to the blind administration of glucose. METHOD: To avoid the confounding effects of a current eating disorder, and to assess possibly trait-related disturbances, we studied 10 subjects recovered (> or = 1 year) from a bulimic-type eating disorder and 6 control women. Subjects were administered a solution of glucose or artificial saliva (control solution) in alternating blocks during a functional magnet resonance imaging scan. RESULTS: Individuals who recovered from a bulimic type eating disorder had significantly lower activation in the right anterior cingulate cortex (ACC; Montreal Neurological Institute [MNI] coordinates x = 8, y = 22, z = 28; cluster size = 18 voxels, T = 5.11, Z-score = 3.78) and in the left cuneus (occipital cortex; MNI coordinates x = -12, y = -78, z = 10; cluster size = 21 voxels, T = 4.27, Z-score = 3.36), when glucose was compared with artificial saliva. CONCLUSION: The ACC plays a role in the anticipation of reward. Individuals with bulimic-type eating disorders may have a reduced reward response to nutrients, and thus may be vulnerable to overeating. However, this is a small sample and the current study will need replication in a larger sample size with investigation of additional regions of interest. PMID- 16254869 TI - Dissonance-based eating disorder prevention program: a preliminary dismantling investigation. AB - OBJECTIVE: A dissonance-based program aimed at reducing thin-ideal internalization has been found to significantly decrease levels of bulimic symptoms in young adult and adolescent females. Because this program is multifaceted, containing psychoeducation, counterattitudinal advocacy, and behavioral exposure components, the current study sought to investigate the mechanisms involved in symptom reduction. METHOD: The current study compared the original treatment program with a dismantled version of the full package, which consisted solely of the specific dissonance component (i.e., the counterattitudinal advocacy procedure). Seventy-eight women were randomly assigned to either the full treatment condition or the counterattitudinal advocacy condition. RESULTS: Findings suggest that both interventions significantly reduced established risk factors for eating pathology as well as bulimic symptoms at termination and at 1-month follow-up. CONCLUSION: Both treatments appear to be equally effective at reducing eating pathology in at-risk college women. Limitations of the study are discussed, and directions for future research are offered. PMID- 16254870 TI - Comorbidity of body dysmorphic disorder and eating disorders: severity of psychopathology and body image disturbance. AB - OBJECTIVE: The current study examined comorbidity and clinical correlates of eating disorders in a large sample of individuals with body dysmorphic disorder (BDD). METHOD: Two hundred individuals with DSM-IV (4th ed. of the Diagnostic and Statistical Manual of Mental Disorders. Washington, DC: American Psychiatric Association; 1994) BDD completed reliable interviewer-administered and self report measures, including diagnostic assessments and measures of body image, symptom severity, delusionality, psychosocial functioning, quality of life (QOL), and history of psychiatric treatment. RESULTS: A total of 32.5% of BDD subjects had a comorbid lifetime eating disorder: 9.0% had anorexia nervosa, 6.5% had bulimia nervosa, and 17.5% had an eating disorder not otherwise specified. Comparisons of subjects with a comorbid lifetime eating disorder (n = 65) and subjects without an eating disorder (n = 135) indicated that the comorbid group was more likely to be female, less likely to be African American, had more comorbidity, and had significantly greater body image disturbance and dissatisfaction. There were no significant group differences in BDD symptom severity, degree of delusionality, or suicidal ideation or attempts. Functioning and QOL were notably poor in both groups, with no significant between-group differences. However, a higher proportion of the comorbid eating disorder group had been hospitalized for psychiatric problems. This group had also received a greater number of psychotherapy sessions and psychotropic medications. CONCLUSION: Eating disorders appear relatively common in individuals with BDD. BDD subjects with a comorbid eating disorder differed on several demographic variables, had greater comorbidity and body image disturbance, and had received more mental health treatment than subjects without a comorbid eating disorder. These findings have important implications for the assessment and treatment of these comorbid body image disorders. PMID- 16254871 TI - Time trends in eating disturbances in young Greek migrants. AB - OBJECTIVE: The study intends to measure time trends in eating disorder psychopathology in Greek adolescents in Veria (Greece) and migrant Greek adolescents in Munich (Germany). For this purpose, large samples of students were assessed at both locations in the 1980s and about two decades later. Our research question was whether the frequency of eating disorder-related psychopathology had changed over time and that there were differences between migrants and nonmigrants. The present-day prevalence of eating disorders in the Greek population was established. METHOD: Greek adolescents were assessed in Munich and Veria in the 1980s (N = 2,631) and almost two decades later (N = 2,920). At both times, the Anorexia Nervosa Inventory for Self-Rating (ANIS) was used to assess eating disorder pathology and the General Health Questionnaire (GHQ-28) was used to assess mental health status. In the second wave, persons at risk for an eating disorder were interviewed using the Structured Interview for Anorexic and Bulimic Syndromes (SIAB-EX). RESULTS: At both times and both locations, adolescent girls in comparison to boys had higher, more pathologic scores on the GHQ-28 and on all ANIS self-rating subscales. Females in Munich reported an increase over time in figure consciousness and their fear of negative effects of meals. In the 1980s, significantly higher scores of bulimic behavior were found in Veria as compared with Munich. In the second wave, bulimic behavior was considerably decreased in Veria for both girls and boys, and for bulimic behavior no significant differences were found between locations. The percentage of girls with a low body weight (<5th percentile) increased significantly over time in Veria and Munich. In the second wave, the current prevalence for girls with anorexia nervosa was 0.00% in Munich and 0.59% in Veria (lifetime 1.26% and 1.18%, respectively). For bulimia nervosa, current prevalence was 1.89% in Munich and 1.18% in Veria. CONCLUSION: Differences between locations diminished over time. Bulimic syndromes are prevalent in both locations. PMID- 16254872 TI - Travel distance and attrition in outpatient eating disorders treatment. AB - OBJECTIVE: The purpose of this study was to investigate the impact of the distance patients had to travel for treatment on attendance patterns and treatment attrition. METHOD: Contact information, clinical records, and/or appointment records of 209 adult patients presenting to an outpatient eating disorder treatment center over a specific period of time were reviewed. Information was obtained on demographics, diagnosis, number of appointments attended, cancelled, and failed, and termination status. Patients were classified as treatment completers or dropouts and compared on demographic, diagnostic, attendance, and distance to treatment site variables. RESULTS: Treatment completer and dropout groups did not differ significantly on demographic variables, with the exception of employed patients being more likely to drop out of treatment. Although not statistically significant, patients diagnosed with bulimia nervosa (BN) and eating disorder not otherwise specified (EDNOS) were more likely to drop out of treatment prematurely. Surprisingly, distance traveled to the treatment site was not significantly different between groups and did not appear to significantly impact attendance patterns. CONCLUSION: Results of this archival investigation were unexpected and likely limited by the design. Results can be useful in understanding motivational factors inherent in noncompliance and premature termination of treatment. A prospective study including fine-grained analysis of variables associated with eating disorder treatment attrition is indicated. PMID- 16254873 TI - Early response to desipramine among women with bulimia nervosa. AB - OBJECTIVE: Numerous trials have demonstrated the efficacy of antidepressant medications for the treatment of bulimia nervosa (BN). The current study examined whether early response to medication predicted response to medication at the end of a controlled trial. METHOD: Data from two previously published studies of desipramine (DMI) were used. Seventy-seven patients with BN were included in the analysis. Receiver operating characteristic (ROC) curves were constructed to examine the relation between the percentage reduction in symptoms at each week and failure to respond to antidepressant medication at the end of the trial. RESULTS: Eventual nonresponders to DMI could be reliably identified in the first 2 weeks of treatment. CONCLUSION: The current study provides preliminary evidence that patients with BN who will not respond to antidepressant medication can be identified in the first 2 weeks of treatment. PMID- 16254874 TI - Ultrasound biometry evolves in the management of diabetes in pregnancy. PMID- 16254875 TI - Ultrasound features of different histopathological subtypes of borderline ovarian tumors. AB - OBJECTIVE: To describe the gray-scale sonographic and color Doppler imaging features of the most common histopathological subtypes of borderline ovarian tumors. METHODS: We analyzed retrospectively the preoperative transvaginal sonographic reports of patients with a histological diagnosis of borderline ovarian tumor. All patients were scanned consecutively by two of the investigators using transabdominal and transvaginal gray-scale imaging to assess the morphology and color Doppler to obtain indices of the blood flow. Sonographic findings were compared to histopathological data. RESULTS: A total of 113 consecutive cases were reviewed from two referral centers for gynecological oncology. At histological examination 50 tumors (44%) were classified as being serous borderline ovarian tumors (SBOT), 61 (54%) were mucinous borderline ovarian tumors (MBOT) (42 intestinal type and 19 endocervical type), and two patients (2%) presented with borderline endometrioid tumors. SBOTs and endocervical-type MBOTs had very similar sonographic features and a smaller diameter, fewer locules (usually unilocular-solid lesions) and a higher color score than intestinal-type MBOTs. Intestinal-type MBOTs were characterized by a significantly higher percentage of lesions with > 10 locules when compared with the endocervical-type MBOTs. CONCLUSION: Intestinal-type MBOTs have different sonographic features from other common borderline ovarian tumors. PMID- 16254876 TI - The use of Z-scores in the analysis of fetal cardiac dimensions. PMID- 16254877 TI - Extracerebellar ectopic brain tissue in the posterior fossa. PMID- 16254878 TI - Development of Z-scores for fetal cardiac dimensions from echocardiography. AB - OBJECTIVES: Z-scores for cardiac dimensions are well established in postnatal life, but have yet to be developed for fetal cardiac dimensions. These would be of real advantage to the clinician in accurately quantifying size and growth of cardiac dimensions and to the researcher by allowing mathematical comparison of growth in differing subgroups of a disease. The purpose of this observational study, conducted at tertiary fetal medicine and cardiology units, was to produce formulae and nomograms allowing computation of Z-scores for fetal cardiac dimensions from knowledge of femur length (FL), biparietal diameter (BPD) or gestational age (GA) using fetal echocardiography. METHODS: Seventeen fetal cardiac dimensions were measured in 130 pregnant women with singleton fetuses of gestational age 15-39 weeks. Regression equations were derived relating all dimensions to FL, BPD and GA. From the calculations, formulae were then developed allowing fetal cardiac Z-score computation. RESULTS: The relationships between cardiac dimensions and FL, BPD or GA were described following natural log transformation. From this analysis, FL (taken as an expression of fetal size) had the highest correlation to fetal cardiac dimensions. From the developed nomograms, Z-scores of specific fetal cardiac structures could be estimated from knowledge of the FL, BPD or GA and echocardiographically derived measurements. CONCLUSIONS: This study allowed computation of Z-scores in fetal life for 17 cardiac dimensions from FL, BPD or GA. Previous studies of normal data allowed qualitative assessment of where abnormal cardiac dimensions lay with regard to the normal range. Z-scores from this study allow quantitative analysis of where such dimensions lie relative to the mean. This permits exact assessment of growth of fetal cardiac structures in normal hearts and particularly in congenitally abnormal hearts where quantitative assessment of the growth of cardiac structures is important in analyzing and planning treatment strategies. PMID- 16254879 TI - Fetal cardiac contractility before and after intrauterine transfusion. AB - OBJECTIVE: To evaluate the effect of fetal anemia and intrauterine transfusion on ventricular shortening fraction. METHODS: The end-diastolic and end-systolic transverse dimensions of the left and right ventricles were obtained using M-mode ultrasonography. The shortening fractions of both ventricles were calculated at three time points: before, immediately after and one day after intrauterine transfusion. The blood volume given at intrauterine transfusion was expressed as a percentage of estimated fetoplacental blood volume. RESULTS: Complete measurements were obtained from 49 transfusions in 23 fetuses. Intrauterine transfusion was performed at a median gestational age of 31 (range, 19-35) weeks. Median hemoglobin concentration before and after intrauterine transfusion was 7.9 (range, 2.7-13.7) g/dL and 14.3 (range, 12.7-16.1) g/dL, respectively. Both left and right ventricular shortening fractions differed significantly between the three time points. Left ventricular shortening fraction decreased immediately after transfusion in 43 (88%) of the 49 procedures. Right ventricular shortening fraction decreased immediately after transfusion in 42 (86%) of the 49 procedures. At the first intrauterine transfusion, there was only a weak correlation between the decrease in shortening fraction of both ventricles and the transfused volume (left: R(2) = 0.15; P = 0.20/right: R(2) = 0.005; P = 0.81). CONCLUSION: Transfusion significantly decreases the shortening fraction of both ventricles of the fetal heart. There is, however, little correlation between the decrease in shortening fraction and the volume of red cells given at intrauterine transfusion. Published by John Wiley & Sons, Ltd. PMID- 16254880 TI - Inversion mode spatio-temporal image correlation (STIC) echocardiography in three dimensional rendering of fetal ventricular septal defects. PMID- 16254881 TI - The fetal cerebellar vermis: anatomy and biometric assessment using volume contrast imaging in the C-plane (VCI-C). AB - OBJECTIVES: To describe the normal appearance and study the biometry of the fetal cerebellar vermis by three-dimensional (3D) volume contrast imaging in the coronal (C-) plane (VCI-C). METHODS: A total of 203 normally developed fetuses were examined prospectively at 18-33 weeks' gestation. At the level of the view used to measure the transverse cerebellar diameter (TCD), a VCI-C plane was displayed to examine, using a transabdominal probe, the fetal mid-saggital vermis. The volumes acquired were stored for later review and measurement of the anteroposterior (AP) diameter, craniocaudal (CC) diameter and surface area of the cerebellar vermis. Each dataset was evaluated by two independent observers. Measurements as a function of gestational age (GA), biparietal diameter (BPD), head circumference (HC) and TCD were expressed by regression equations. Interobserver variability was evaluated. Nomograms were produced. In order to validate the use of VCI in fetal biometry, datasets from 57 patients were selected arbitrarily for comparison of their VCI-C measurements with those from mid-sagittal sections of a stored 3D multiplanar examination. Intraclass correlation was used to evaluate the agreement between these measurements. RESULTS: The mean maternal age was 32 years. We were able to measure mid-sagittal CC diameter, mid-sagittal AP diameter and cerebellar vermis surface area in all fetuses. Interobserver variability analysis showed no significant differences between the two observers (P > 0.05). Measurements of the cerebellar vermis (AP diameter, CC diameter and surface area) correlated linearly with GA, BPD, HC and TCD (r > or = 0.82, P < 0.0001). CC and AP diameters estimated from the mid sagittal section of the multiplanar measurements were significantly correlated with VCI-C measurements (r = 0.96, P < 0.00001 and r = 0.95, P < 0.00001, respectively). CONCLUSIONS: VCI-C is a valuable tool, allowing intrauterine assessment of the normal appearance of the fetal cerebellar vermis. The nomograms developed in this study should enable accurate evaluation of the cerebellar vermis. PMID- 16254882 TI - Cord entanglement of monochorionic diamniotic twins following spontaneous antepartum septostomy sonographically simulating a true knot of the umbilical cord. AB - Spontaneous antepartum septostomy occurring in monochorionic diamniotic twins is extremely rare. We present a case in which prenatal sonography at 26 weeks' gestation depicted a monochorionic diamniotic twin gestation with concordant fetal growth and findings suggestive of a true knot of the umbilical cord. At Cesarean delivery at 34 weeks' gestation, spontaneous antepartum septostomy with entanglement of the two separate umbilical cords was noted. This case suggests that the differential diagnosis of findings considered consistent with a true knot of the umbilical cord in monochorionic diamniotic twin gestations, should include spontaneous antepartum septostomy and umbilical cord entanglement. PMID- 16254883 TI - The use of high resolution magnetic resonance imaging in the prenatal diagnosis of fetal nuchal tumors. AB - Fetal magnetic resonance imaging (fetal MRI) is an important adjunct to antenatal imaging especially when neonatal surgery is contemplated. We report two cases of fetal nuchal tumors, which were diagnosed incidentally on an ultrasound scan and had fetal MRI to aid diagnosis, prognosis, counseling and management planning. In the first case, fetal MRI aided diagnosis and prenatal multidisciplinary management of a cervical teratoma. Tracheal involvement could not be excluded and an ex-utero intrapartum treatment procedure was planned. Postnatal MRI and angiography provided further information prior to surgery. In the second case, fetal MRI assisted thorough counseling following the finding of a cervical lesion thought to be a cervical teratodermoid, a multicystic hygroma or congenital lymphectasia. The parents opted for termination of the pregnancy. Postmortem findings confirmed the extent of involvement of surrounding structures diagnosed prenatally. The mass was found to be a hamartomatous hemangiolymphangioma. PMID- 16254884 TI - Z-velocity in screening for intrauterine growth restriction. AB - OBJECTIVES: Ultrasound scans provide the basis for detection of intrauterine growth restriction (IUGR) but often fail to distinguish IUGR from small-for gestational age (SGA) fetuses. This study introduces the concept of Z-velocity, calculated as changes in Z-scores over time, as an additional criterion in the diagnosis of IUGR. METHODS: A computer program simulated 50 000 fetal abdominal circumference (FAC) scans based on published growth formulae. False-positive rates were calculated to determine optimal scan time and scan intervals. Using an independent simulation of 32 500 FAC scans, the two methods were compared using receiver-operating characteristics (ROC) curve analysis. RESULTS: ROC showed areas under the curve of > 0.74 over the complete range of scan intervals. The positive predictive value of growth arrest as the only diagnostic criterion was, however, too low to recommend it as an exclusive or the first diagnostic criterion. CONCLUSIONS: Z-velocity can be used to decide whether further investigations for growth abnormality are required in fetuses that fall below the 10(th) percentile. The gain of combined diagnostic approaches should be calculated from large databases that include the neonatal ponderal index as the gold standard. PMID- 16254885 TI - Free radical scavenging and antigenotoxic activities of natural phenolic compounds in dried flowers of Hibiscus sabdariffa L. AB - The antioxidant and free radical scavenging effects of two fractions of the ethanolic extract (HSCF, chloroform soluble fraction and HSEA, ethyl acetate soluble fraction) obtained from the dried flowers of Hibiscus sabdariffa L were investigated. The total antioxidant activity of the extracts was estimated to be 4.6 and 8.6 mM of vitamin C for HSCF and HSEA, respectively. Both HSCF and HSEA scavenged hydrogen peroxide (H(2)O(2)) (79-94%) at the dose of 500 microg. Similarly, the extracts showed inhibitory (70-80%) effects on superoxide anions radicals (O(2) (- *)) at a dose of 1000 microg. The concentrations required for a 50% scavenging of hydroxyl radical (OH) (IC(50)) were 380 and 200 microg for HSCF and HSEA, respectively. HSEA and HSCF were better scavengers of O(2) (- *), *OH and H(2)O(2) as compared to BHA, quercetin and alpha-tocopherol. At a concentration of 25 microg/mL HSCF and HSEA exhibited 32 and 38% inhibition on CCl(4)-NADPH-induced lipid peroxidation, respectively, while both extracts exhibited 80 and 89% inhibitory effects at 100 microg/mL. Pretreatment with H. sabdariffa extracts orally with 100 mg/kg and 250 mg/kg simultaneously with intraperitoneal injection FeCl(2)-ascorbic acid-ADP mixture reduced (p < 0.01) the formation of malondialdehyde content. Treatment of rats with HSCF, HSEA and vitamin C (standard antioxidant) significantly inhibited the induction of micronucleated polychromatic erythrocytes by sodium arsenite (2.5 mg/kg) (p < 0.001) after 24 h by 60, 70 and 50%, respectively. The results indicate that extracts of H. sabdariffa showed strong antimutagenic activity and free radical scavenging effects on active oxygen species. PMID- 16254886 TI - Electron spin resonance spectroscopy studies on the free radical scavenging activity of wine anthocyanins and pyranoanthocyanins. AB - Anthocyanins are a group of natural occurring pigments responsible for the red blue color of grapes and many fruits and vegetables. Anthocyanins and derived pigments are of double interest, one technological, as they can be used as natural colorants, and another one due to their implication on human health through their antioxidant activity. Although there are numerous studies regarding the antioxidant activity of grape extracts as well as red wine, the free radical scavenging activity of purified anthocyanins and pyranoanthocyanins is largely unknown. In the present study, the hydroxyl and superoxide anion scavenging activities of anthocyanins and their pyruvic acid adducts were systematically investigated by electron spin resonance spectroscopy and spin trapping. The 3 glucosides of delphinidin, cyanidin, petunidin, pelargonidin and malvidin, and the pyruvic adduct of the 3-glucoside of delphinidin exhibited a potent superoxide anion radical scavenging and, to a lesser extent hydroxyl anion radical scavenging activity. The pyranoanthocyanins of cyanidin, petunidin, malvidin and pelargonidin showed a high capacity to scavenge superoxide anion radicals but did not scavenge hydroxyl radicals. Current data indicate that formation of anthocyanin adducts with pyruvic acid, which may occur during wine ageing or fruit juice processing, decreases the hydroxyl and superoxide anion scavenging and thus could decrease the antioxidant potential of these compounds. PMID- 16254887 TI - Suppression of arachidonic acid metabolism and nitric oxide formation by kudzu isoflavones in murine macrophages. AB - Inhibitory effect of kudzu isoflavones on arachidonic acid metabolism and nitric oxide (NO) production in lipopolysaccharide activated RAW 264.7 macrophages were investigated. Isoflavone aglycones, such as daidzein, genistein, biochanin A, and formononetin significantly suppressed arachidonic acid release (50 microM). Biochanin A, which displayed the most active inhibition on arachidonic acid release in HT-29 human colon cancer cells, exhibited its most potent suppression in RAW 264.7 cell (by 86%) without showing cytotoxicity. However, isoflavone glucosides, puerarin and daidzin, showed lower inhibitory activities on the release of arachidonic acid and its metabolites. In NO formation, biochanin A showed marked inhibition, by 62% (50 microM), followed by genistein, daidzein, formononetin, and daidzin, 56, 39, 33, and 8%, respectively. 5,7-Dihydroxyl group in the A-ring of isoflavones could be a key functional group responsible for the strong inhibitory activity of biochanin A and genistein on NO production. These activities may contribute to the antiinflammatory and anticarcinogenic properties of kudzu isoflavones. PMID- 16254888 TI - Antioxidant activity of pasteurized and sterilized commercial red orange juices. AB - Blood orange juice is a typical Italian product whose red color is primarily associated with anthocyanin pigments. Two orange-based products are present on the market: pasteurized pure juice with 40 days of shelf life, and sterilized beverage containing minimum 12% of concentrated fruit juice. The aim of the present paper is to verify the relationships between the antioxidant properties and the anthocyanins content in a sampling of pasteurized and sterilized commercial red orange juices. The anthocyanins composition was determined by HPLC MS/MS, while the antioxidant activity was evaluated by the Briggs-Rauscher reaction, selected in order to acquire information at acid pH values, by three radical scavenging assays (DMPD, 2-2'-azinobis-(3-ethylenbenzothiazoline-6 sulfonic acid) diammonium salt (ABTS), DPPH), and by FRAP assay to monitor the ferric reducing power. Results showed that antioxidant activity, particularly when measured by ABTS method, is positively related to the content of anthocyanins and that the reduction of anthocyanins content, typical of commercial long-shelf life juices, leads to a remarkable loss of antioxidant power. PMID- 16254889 TI - Prenatal diagnosis of fetal bilateral adrenal carcinoma. AB - Fetal malignancies that are capable of metastasizing to other fetal organs and the placenta are exceedingly rare. Fetal tumors are suspected on ultrasonography in the presence of structures of abnormal shape or size that are occasionally associated with polyhydramnios and hydrops. Most of the large abdominal masses detected antenatally are renal in origin, although adrenal tumors should also be kept in mind in their differential diagnosis. We report a case of a large-for gestational-age fetus with abdominal distention secondary to bilateral adrenal carcinoma, polyhydramnios and placental enlargement. Postmortem histological findings included nesidioblastosis, (i.e. hyperplasia of the cells of the islets of Langerhans) and adrenocortical cytomegaly, suggestive of a form of Beckwith Wiedemann syndrome. PMID- 16254890 TI - The amniotic sheet: a truly benign condition? AB - OBJECTIVE: Amniotic sheets are the result of uterine synechiae that have been encompassed by the expanding chorion and amnion. Radiologically they are seen as 'shelves' in the amniotic cavity. The benign nature of such amniotic sheets has been documented in many case series in the literature. The objective of this study was to determine the characteristics (if any) of amniotic sheets that predict fetal outcome. METHODS: Between January 2001 and December 2002, detailed scans were performed in 30 476 singleton pregnancies at 18-32 weeks' gestation. Of these, 44 cases of amniotic sheets were detected. The characteristics studied were site of amniotic sheet and whether the amniotic sheet was complete (i.e. no free edge seen on ultrasound) or incomplete (i.e. presence of free edge seen on ultrasound). The primary fetal outcome studied was stillbirth. RESULTS: The incidence of amniotic sheets was 0.14%. Two were complete and 42 were incomplete. Of the 38 cases with known outcomes there were two intrauterine deaths. There was no association between fetal outcome and the uterine location of the amniotic sheet (i.e. upper two-thirds vs. lower third, P = 0.5). There was, however, an association between the completeness of the amniotic sheets and intrauterine death (P = 0.002). Both instances of intrauterine death occurred in the two cases with complete amniotic sheets. Postmortem examination suggested that cord accidents were the cause of intrauterine death in both cases. CONCLUSIONS: This study supports the view that incomplete amniotic sheets are benign. It also suggests for the first time that complete amniotic sheets may be associated with intrauterine death. PMID- 16254891 TI - Successful vesicoamniotic shunting of posterior urethral valves in the first trimester of pregnancy. AB - Prenatal ultrasonography early in gestation allows diagnosis of posterior urethral valves. We report on a fetus with posterior urethral valves treated using vesicoamniotic shunting at 13 + 5 weeks' gestation. A double-basket catheter was used for shunting. A 2582-g male neonate was delivered at 33 weeks' gestation, and the infant continued to show normal renal function at 3 years of age. To the best of our knowledge this is the first report of successful vesicoamniotic shunt placement using a double-basket catheter in the first trimester of pregnancy. PMID- 16254892 TI - Four-dimensional ultrasound guidance of prenatal invasive procedures. AB - OBJECTIVE: Technological advances in ultrasonography have revolutionized prenatal diagnosis and treatment. Here we evaluate the effectiveness of using four dimensional (4D) ultrasonography to guide prenatal invasive procedures. PATIENTS AND METHODS: Prenatal invasive procedures using 4D ultrasound were recorded prospectively in 93 cases: 10 amnioinfusions, 50 amniocenteses, 8 chorionic villus samplings (CVS) and 25 cordocenteses. The needle target site was first identified using the two-dimensional (2D) mode, and was then confirmed using the three-dimensional (3D) mode. The needle was inserted under 4D ultrasound guidance. After selecting the needle target site, the true position of the needle was determined in three planes ('real-time 3D targeting'). RESULTS: Using 4D ultrasound guidance, most procedures were performed within 5 min and with a 100% success rate, even in cases involving severe oligohydramnios (amniocentesis), thin placentas (CVS) or narrow umbilical veins (cordocentesis). Moreover, there were no serious complications during or after any procedure. CONCLUSIONS: 4D ultrasonography can be used to guide various prenatal invasive procedures offering clear information in all three planes. It is likely that such imaging will reduce the time taken to complete the procedures and reduce the risks associated with them. Published by John Wiley & Sons, Ltd. PMID- 16254893 TI - The aging brain: the cognitive reserve hypothesis and hominid evolution. AB - Compared to other primates, humans live a long time and have large brains. Recent theories of the evolution of human life history stages (grandmother hypothesis, intergenerational transfer of information) lend credence to the notion that selection for increased life span and menopause has occurred in hominid evolution, despite the reduction in the force of natural selection operating on older, especially post-reproductive, individuals. Theories that posit the importance (in an inclusive fitness sense) of the survival of older individuals require them to maintain a reasonably high level of cognitive function (e.g., memory, communication). Patterns of brain aging and factors associated with healthy brain aging should be relevant to this issue. Recent neuroimaging research suggests that, in healthy aging, human brain volume (gray and white matter) is well-maintained until at least 60 years of age; cognitive function also shows only nonsignificant declines at this age. The maintenance of brain volume and cognitive performance is consistent with the idea of a significant post- or late-reproductive life history stage. A clinical model, "the cognitive reserve hypothesis," proposes that both increased brain volume and enhanced cognitive ability may contribute to healthy brain aging, reducing the likelihood of developing dementia. Selection for increased brain size and increased cognitive ability in hominid evolution may therefore have been important in selection for increased lifespan in the context of intergenerational social support networks. PMID- 16254894 TI - No difference in serum leptin concentrations between urban-dwelling Austronesians and Non-Austronesians in Papua New Guinea. AB - Pacific Islands populations can be broadly divided into Austronesians (AN) and Non-Austronesians (NAN); obesity and type 2 diabetes are prevalent in the former, although leptin levels in both groups have seldom been investigated. Thirty-seven (20 male and 17 female) adult pairs, matched by age and percent body fat, from AN speaking Balopa and NAN-speaking Huli, all of whom migrated to settle in Port Moresby, the capital of Papua New Guinea, were selected for comparison of their serum leptin concentrations. The Balopa did not differ significantly from the Huli in age (30.5 +/- 9.7 and 30.0 +/- 8.7 years for males, 33.7 +/- 8.9 and 34.1 +/- 7.5 years for females, respectively) or percent body fat (19.4 +/- 5.6 and 18.8 +/- 4.6 for males, 34.1 +/- 6.2 and 33.3 +/- 5.0 for females), although the BMI of females was lower in the Balopa (26.4 +/- 4.9) than in the Huli (29.7 +/- 4.7) (P = 0.02). In both ethnic groups, females had markedly higher leptin concentrations than males, but there was no significant inter-group difference in males (3.5 +/- 2.6 and 3.1 +/- 4.7 ng/ml, P = 0.14) or females (22.7 +/- 12.9 and 19.7 +/- 11.9 ng/ml, P = 0.40), after controlling for lifestyle factors and serum lipids. Multiple regression analysis revealed that significant predictors of leptin concentration were % body fat (beta = 0.58), sex (male, 0; female, 1; beta = 0.27), and smoker status (non-smoker, 0; smoker, 1; beta = -0.15) (R(2) = 0.80), implying that the leptin concentration was primarily determined by lifestyle-derived body fatness. In conclusion, the NAN populations do not endogenously differ in leptin status from the AN populations, who have been recognized as a typical group with a "thrifty" genotype. PMID- 16254895 TI - Longitudinal models of long bone growth during adolescence. AB - Data from the Child Research Council (Denver, CO) were utilized to model longitudinal adolescent growth of the humerus, radius, femur, and tibia for 36 girls (10-16 years) and 33 boys (10-17 years). Multilevel modeling procedures were used to estimate variation, covariation, and the polynomial parameters necessary for generating growth curves. At age 10, long bone lengths for girls and boys are similar; by age 16, each of the boys' arm bones is about 20 mm longer and each of their leg bones is about 30 mm longer. Due to the earlier maturation of girls, the models show the length of each of their long bones exceeding that of boys to some degree during some period of adolescence. Peak velocities for leg bones are attained earlier than those for arm bones; in both sexes, age at humeral peak velocity coincides with age at peak height velocity (PHV). At age 13, correlations among lengths and among velocities of bones are strong to moderate, and girls consistently display higher variation than do boys for both long bone length and growth velocity. Considered relative to the average velocity of each bone's growth, the tibia is the most variable of the four long bones at age 13 years. PMID- 16254896 TI - Methodological issues in the anthropometric assessment of Hmong children in the United States. AB - The effectiveness of the body mass index (BMI; kg/m2) in assessing overweight/obesity may be diminished in populations of short stature. In a sample (n = 79) of Hmong refugee children in the United States, of age 4-11 years, median z scores for height, BMI, and the triceps skinfold were -1.04z, +0.53z, and +0.18z, respectively. Further, 41.7% of children were above the NHANES 85th percentile for BMI-for-age, categorizing them as overweight/obese. Assessment of obesity by other established criteria for children, such as the triceps skinfold and body fat percentage, produced significantly lower estimates than did BMI. This is consistent with patterns found in other stunted populations, suggesting that BMI be employed in conjunction with other methods when assessing overweight/obesity in these groups. Finally, although stunting and overweight/obesity were both common in this study, at the individual level height z scores were positively correlated to z scores of various measures of adiposity. PMID- 16254898 TI - Proceedings of the Human Biology Association 30th Annual Meeting, April 5-6, 2005, Milwaukee Hilton, Milwaukee, WI. PMID- 16254897 TI - The second to fourth digit ratio in elite and non-elite female athletes. AB - Contests in the animal world to determine social status almost exclusively involve males, which points out that androgens may be indispensable in the development of competitive instincts. In animal studies, it has been shown that prenatal exposure to androgens may produce permanent changes toward more aggressive behavior in adulthood. Thus, there is a strong suspicion that women involved in competitive activities, such as sports, may have been exposed to high androgen levels in utero. There is strong evidence that the ratio between the second to fourth digits ratio (2D:4D ratio) correlates negatively with intrauterine androgen concentrations and could potentially be used as a marker for prenatal androgen exposure. Therefore, the purpose of our study was to test the hypothesis that women engaged in sports have lower 2D:4D ratio-a marker of high prenatal androgen exposure. We measured the 2D:4D ratios in elite and non elite female athletes and compared them with female individuals not engaged in any sport activities. Our results showed that elite female athletes have significantly lower left hand 2D:4D ratios compared to the control group (P < 0.05). Therefore, we can speculate that low 2D:4D ratio may be a positive correlate of sports potential in females. PMID- 16254899 TI - An unexpected wide population variation of the G1733A polymorphism of the androgen receptor gene: data on the Mediterranean region. AB - The androgen receptor (AR) has been proposed as a candidate gene for several cancers (breast, prostate, uterine endometrium, colon, and esophagus). Ethnicity is considered an associated risk factor for some of these cancers. Several case control genetic studies have been focused in samples of the main ethnic groups, but little is known about the distribution of risk polymorphisms in current populations with accurate ethnic and/or geographic origins. The A allele of the G1733A polymorphism of the AR gene has been associated with increased risk of prostate cancer. We provide data from this marker in 12 samples from 7 Mediterranean countries such as Spain, Italy (Sardinia), Greece, Turkey, Morocco, Algeria, and Egypt. A sample from Ivory Coast has also been analyzed. The A allele distribution shows a frequency in the Ivory Coast population (65.17%) that contrasts with the low values found in Northern Mediterraneans (mean average value of 13.98%). North African populations present two-times higher frequencies (average value of 27.19%) than Europeans. The wide population variation range found for the A allele strengthens the potential interest of further screening as a baseline to the design of future preventive and population health programs. PMID- 16254900 TI - Cross-sectional study of echocardiographic characteristics in healthy children living at high altitude. AB - Non-echocardiographic studies in healthy high altitude children have shown right ventricle predominance during infancy and childhood, associated to asymptomatic pulmonary hypertension and an increased pulmonary artery pressure. Systematic studies on echocardiography in such children have not been performed. In a cross sectional study, we measured right and left heart morphologic and functional parameters, through M-mode, two-dimensional Doppler, and color Doppler echocardiographies, in a population of 321 healthy children ranging in age from 2 months to 19 years and living at high altitude (Tintaya, Peru, 4,100 m). Structured ad-hoc interviews were done to obtain information on medical history, patterns of exposure to high altitude of children and their parents and grandparents, place and altitude of pregnancy and birth, and housing conditions. A complete physical examination was performed before echocardiography. Hemoglobin concentration, pulse oximetry, and anthropometry were measured in all participating children. The right and left heart morphologic and functional echocardiographic measurements expressed by age and by body surface area were generally similar to sea-level reference populations. They were not consistently influenced by sex, nutritional status, chest dimensions, pulse oximetry, hemoglobin concentration, ethnicity, length of residence at high altitude, or parental history of exposure to high altitude. Most children had at least some degree of high-altitude ancestry as assessed by ethnicity and history of parental exposure to altitude. The cardiovascular development at high altitude in children with some degree of high-altitude ancestry seems to follow a pattern similar to sea-level children. The results can be used as reference values to interpret individual echocardiographic studies in comparable children living in similar settings. PMID- 16254901 TI - Longitudinal estimates of puberty timing in Senegalese adolescent girls. AB - This study aimed to assess the timing of sexual maturation (breast development and menarche occurrence) among sub-Saharan African adolescent girls from rural areas. In the framework of a longitudinal study of growth at puberty, the stages of pubertal development (Tanner classification) and menarche occurrence were recorded at intervals between 1995 and 2000 in a sample of 406 Senegalese adolescent girls from a rural area. Nutritional status was estimated during infancy, childhood, and adolescence within this sample, and body composition was estimated only during adolescence. At the beginning of the study (1995), the adolescent girls were 11.4 years old (SD = 0.6). At the end of the study, they were 16.5 years old (SD = 0.6). Median and quartile ages at entry into each maturity stage and its confidence limits were estimated by a lognormal parametric survival model. These adolescent girls were shorter and thinner than girls of the same age from developed countries. Median age at the onset of breast development was 12.6 years (95% CI = 12.5-12.8). Median age at the end of sexual maturation and median age at menarche were estimated respectively at 15.8 years (95% CI = 15.7-15.9) and 15.9 years (95% CI = 15.7-15.9). These adolescent girls were less mature than other adolescent girls of the same age from other developing countries. Their puberty was extremely delayed compared to that of adolescent girls of the same age from industrialized countries. Puberty in these Senegalese adolescent girls is delayed by about 3 years. This may be due to malnutrition, significant energy expenditure, and poor diet and living conditions, and possibly to genetic control. PMID- 16254902 TI - Effects of progressive strength training on the performance of the Functional Reach Test and the Timed Get-Up-and-Go Test in an elderly population from the rural north of Portugal. AB - The purpose of this study was to examine the effect of an intensity progressive strength training program on the performance of two tests related to fall risk: the Functional Reach Test (FRT) and the Timed Get-Up-and-Go Test (TUG). Twenty subjects were randomly included in one experimental group (age 73 +/- 6 years old) and one control group (age 75 +/- 5 years old). Subjects from the experimental group participated in a strength-training program consisting of three sessions each week for a period of 14 consecutive weeks, from 50% to 80% of 1RM. A 2 (group: experimental vs. control) x 2 (pre-post: pre-test vs. post-test) repeated measures ANOVA was carried out on subjects in TUG and FRT performance. Mean FRT results from the pre-test were significantly lower than mean results from post-test. The two main effects were qualified by a significant group x pre post interaction, F(1,18) = 39.23, P < 0.001, identifying markedly different performance profiles. Mean TUG results from the pre-test were significantly lower than mean results from post-test. The two main effects were qualified by a significant group x pre-post interaction, F(1,18) = 50.74, P < 0.001, again identifying markedly different performance profiles. Globally, the obtained results through our progressive strength-training program showed important improvements in functional task performance and in preventing falls. PMID- 16254903 TI - Beta-globin cluster haplotypes in normal individuals and beta(0)39-thalassemia carriers from Sardinia, Italy. AB - Seven polymorphic sites in the beta-globin cluster in association with specific thalassemia mutations were analyzed in a sample from Sardinia, Italy. In order to verify previous works carried out on normal samples (beta(A)/beta(A)) and family studies on beta-thalassemia homozygotes individuals, the haplotype frequencies in both normal individuals (beta(A)/beta(A)) and beta(0)39-thalassemia carriers (beta(A)/beta0) were studied. In our work chromosomes carrying beta(0)39 mutation are characterized by a prevalence of haplotype II (- + + - + + +) (52%) relative to haplotype I (+- - - - + +) (29%), in contrast, among chromosomes with beta(A) the frequency of haplotype I is much greater than that of haplotype II. These data confirm what was found by other authors. Nevertheless, our results disagree with those of previous studies of Sardinians, both in frequencies values and in the numbers of haplotypes identified. Population analysis performed with samples carrying the beta-thalassemic mutation highlighted the peculiarity of Sardinians with respect to other Mediterranean populations. The Corsican population is most similar to the Sardinian population, confirming previous analyses performed with both classical markers and mitochondrial and genomic DNA. PMID- 16254904 TI - Effects of consanguinity on pre-reproductive mortality: does demographic transition matter? AB - The aim of this study was to investigate whether there is an increase on premature deaths due to genetically determined factors at the beginning of a demographic transition. We also analyzed the effects of parental consanguinity on offspring mortality from an epidemiological viewpoint, using parish records for family reconstitution in a Basque population (1800-1990). Among the offspring of unrelated parents, 13.1% died before their first year of life (infant mortality), and 22.8% died before the age of 16 (pre-reproductive mortality). Significant increases in both infant (23.6%) and pre-reproductive (38.5%) deaths were found among the progeny of first cousins or closer relatives, 1C (F > or = 0.0625). The corresponding relative risks of mortality were 1.79 (95% confidence limits: 1.37 2.28) and 1.68 (1.38-2.01), respectively. Estimates of the population attributable risks indicate that 4% of pre-reproductive mortality is ascribable to consanguineous unions, although kinships other than 1C produced only slight increases in offspring mortality. Evidence on the relationship between the demographic transition and the increase in premature deaths due to genetic factors was obtained through a principal component analysis (95.1% of variance accounted for). During the initial stages of the demographic transition, the population experienced substantial elevations in mean family size, natural increase of the population, frequency of close consanguineous matings (1C), and death rate due to congenital anomalies and perinatal diseases. These findings are of interest for the health services of many developing societies in Asia, Africa, and Latin America, which are nowadays immersed in the demographic transition process. PMID- 16254905 TI - Prevalence of metabolic syndrome in two tribal populations of the sub-Himalayan region of India: ethnic and rural-urban differences. AB - This study was undertaken to estimate prevalence of metabolic syndrome in traditional societies in the sub-Himalayan region and to assess the impact of modernization on the risk to the syndrome. Two tribal populations--Toto and Bhutia--with a shared ancestry and habitat were selected. Some Bhutians have adopted a modern lifestyle. The study design permitted assessment of the relative roles of lifestyle and genetic factors in influencing the prevalence of metabolic syndrome. Our study has revealed that metabolic syndrome (or its contributing variables) can be a major health problem, even in traditional rural ethnic groups, indicating that this syndrome is not necessarily a result of modernization or urbanization. Dyslipidemia seems to be the major contributor to metabolic syndrome. Further, our study indicates that genetic factors that adversely affect the levels of such variables have long antiquities in Indian ethnic groups. We find that there is an additional adverse impact of adoption of urban lifestyles (perhaps primarily mediated through dietary changes) on metabolic syndrome. PMID- 16254906 TI - Exploring the relationship between hypoplasia and odontometric asymmetry in Isola Sacra, an imperial Roman necropolis. AB - Anthropological studies reporting odontometric asymmetry values or dental enamel hypoplasia frequencies use these markers as a record of physiological perturbations occurring during dental development. While both markers indirectly suggest the amount of relative stress a population might have experienced, a relationship between the two has been explored only recently in the literature. In this study, we address the possibility of such a relationship in two ways. First, Kendall's tau B correlations test the degree of relationship on the level of the individual between hypoplasia presence/absence (P/A) and severity of hypoplasia appearance (PS) data for the anterior dentition and directional (DA) and fluctuating asymmetry (FA) data for concurrently developing molars pairs. Second, an F-test explores between-group (ranked hypoplastic individuals and non hypoplastic individuals) variance about the mean, expecting the hypoplastic individuals to be more variable. The sample consists of 72 individuals from the Isola Sacra necropolis, which is associated with Portus, the port city of ancient Rome. Results indicated only a very weak predictive relationship between some variables and few significant differences in variation. However, variance follows trends in published literature. Possible explanations for the lack of interaction on the level of the individual include both etiological and genetic susceptibility factors that are significant in and of themselves as they suggest a more complex reading of the hard tissue evidence for stress in archaeological populations. PMID- 16254907 TI - Directional mating and a rapid male population expansion in a hybrid Uruguayan population. AB - The Uruguayan population has been considered as mainly European descent, with a negligible Native American or African contributions. Based on serological and molecular markers, recent studies demonstrate that these two populations had an important influence in the conformation of the present one. To the Northeastern region of Uruguay, a 20% Native American contribution was estimated using autosomal markers and a 62% Native American female origin based on mitochondrial markers. In this paper, we analyze four Y chromosome markers, two biallelic loci (M3 and YAP) and two microsatellites (DYS389I and DYS391), to characterize the male genetic contribution of a sample from the Northeastern city of Tacuarembo. We take different approaches to estimate the origin of male contributions to the population of Tacuarembo; Native American contribution ranges between 1.60% and 8.31%, confirming strong directional mating, which was also detected before with mitochondrial markers. Furthermore, the male population of Tacuarembo presents the characteristic of a population that suffered a bottleneck and a posterior expansion, confirmed using two microsatellite-based statistics to analyze the past population growth; patrilocality and migration could be responsible of those characteristics. PMID- 16254908 TI - Fibrinogen, homocyst(e)ine, and C-reactive protein concentrations relative to sex and socioeconomic status in British young people. AB - This study assesses the prevalence of recently identified coronary heart disease (CHD) risk factors in young people of differing socioeconomic status (SES). From November 2001 through March 2002, 100 boys and 108 girls, of age 12.9 +/- 0.3 years, selected from differing SES were assessed for CHD risk factors. Measurements included fibrinogen (Fg), homocyst(e)ine (Hcy), and C-reactive protein (CRP). Fibrinogen was significantly greater among boys from a higher SES compared with those from a low SES (P < or = 0.05). Differences according to sex (P < or = 0.05) were identified for Fg and CRP. The data indicate the prevalence of recently identified CHD risk factors in this cohort of British schoolchildren. For the purpose of this article, the phrase "young people" embraces both children and adolescents. PMID- 16254910 TI - Transvaginal sonography in the assessment of distal ureteral calculi. AB - OBJECTIVES: To explore the morphological and functional findings of transvaginal sonography (TVS) in the diagnosis and management of distal ureteral calculi. METHODS: We retrospectively reviewed the imaging studies in seven cases of distal ureteral calculi (study group) and 20 cases of female volunteers without urinary tract infection (control group). All 27 patients had undergone TVS for assessment of the lower urinary tract. The variables measured on ultrasound included the presence or absence of distal ureteral calculi, the size and location of a calculus if present, the presence or absence of the ureter jet phenomenon, morphological changes of the ureteral papilla and adjoining tissue, and the height of the ureteral papilla, as well as vascular changes seen on Doppler imaging. RESULTS: In the controls, 80% of the ureteral papillae were triangular and 20% trapezoidal. The average (+/-SD) height of the ureteral papillae was 3.5 +/- 0.7 mm on the right and 3.6 +/- 0.9 mm on the left. There was no statistically significant difference between the heights of the right and left papillae (P = 0.619). In the study group, echogenic stones were identified in all seven patients and a hypoechogenic tubular structure connected to the involved papilla was identified in 6/7 patients. The average height of the involved papilla was 6.7 +/- 1.6 mm. Ureteric jets were visible in all patients in both the study and control groups. CONCLUSION: In patients with distal ureteral calculi, TVS provides a rapid, non-invasive and repeatable means of assessing the morphology and function of the distal ureter. PMID- 16254909 TI - Resting metabolic rate in a sample of girls in Malaysia and England. AB - This cross-sectional study of age matched (10-11 years), pre-menarcheal girls in England and Malaysia investigated the effect of ethnicity on resting metabolic rate (RMR). The children were recruited from schools in Oxford, England, and Kuala Lumpur, Malaysia, and all measurements were conducted in the schools. The Malaysian girls were shorter (143.7 +/- 6.5 cm and 140.1 +/- 5.3 cm (mean +/- SD) for the English and Malaysian girls respectively) and lighter (32.5 +/- 5.3 kg compared with 38.0 +/- 8.7 kg for the English girls) with a smaller fat mass and fat free mass (FFM) than the English girls. Energy expenditure was lower in the Malaysian girls (4555 +/- 531 kJ/day compared with 5178 +/- 688 kJ/day for the English girls). Although a difference in RMR was observed between the two groups, when corrected for body composition the difference was no longer significant. No effect of ethnicity on the relationship of FFM and RMR was shown when the data were analyzed using analysis of covariance. PMID- 16254911 TI - Female precocious puberty, obesity and polycystic-like ovaries. AB - OBJECTIVE: To evaluate the characteristics of obese girls with gonadotropin releasing hormone-dependent precocious puberty with and without polycystic-like ovaries. METHODS: Forty-seven overweight (> 75(th) centile of the Italian reference data) girls with a diagnosis of isosexual precocious puberty underwent auxological analysis, hormonal assay and utero-ovarian sonographic and Doppler evaluation. On the basis of sonography the patients were subdivided into two groups, girls presenting normal ovaries (Group I; n = 31) and those with polycystic-like ovaries (Group II; n = 16). RESULTS: The mean body weight was significantly higher (P = 0.003) in Group II than it was in Group I. In addition, the patients with polycystic-like ovaries fell within our definition of superobese (> or = 97(th) centile of the Italian reference data) in 44% of cases. The uterine and ovarian volumes were significantly greater in Group II compared with Group I patients. The Doppler evaluation showed intraparenchymal ovarian vascularization and low downstream impedance to flow in all patients in Group II. CONCLUSIONS: Girls with precocious puberty and polycystic ovaries, compared with those without polycystic ovaries, have a higher incidence of body weight exceeding the 85(th) centile of the Italian reference data (obesity). PMID- 16254912 TI - Correspondence of function and phylogeny of ABC proteins based on an automated analysis of 20 model protein data sets. AB - Using our BLAST-based procedure RiPE (Retrieval-induced Phylogeny Environment), which automates the evolutionary analysis of a protein family, we assembled a set of 1138 ABC protein components [adenosine triphosphate (ATP)-binding cassette and transmembrane domain] from the protein data sets of 20 model organisms and subjected them to phylogenetic and functional analysis. For maximum speed, we based the alignment directly on a homology search with a profile of all known human ABC proteins and used neighbor-joining tree estimation. All but 11 sequences from Homo sapiens, Arabidopsis thaliana, Drosophila melanogaster, and Saccharomyces cerevisiae were placed into the correct subtree/subfamily, reproducing published classifications of the individual organisms. By following a simple "function transfer rule", our comparative phylogenetic analysis successfully predicted the known function of human ABC proteins in 19 of 22 cases. Three functional predictions did not correspond, and 10 were novel. Predictions based on BLAST alone were inferior in five cases and superior in two. Bacterial sequences were placed close to the root of most subtrees. This placement coincides with domain architecture, suggesting an early diversification of the ABC family before the kingdoms split apart. Our approach can, in principle, be used to annotate any protein family of any organism included in the study. PMID- 16254913 TI - Aspects of sound communication in the pearlfish Carapus boraborensis and Carapus homei (Carapidae). AB - Several species of Carapidae are known to have symbiotic relationships with marine invertebrates. The two most common species in Moorea (French Polynesia), Carapus boraborensis and Carapus homei, undergo conspecific and heterospecific encounters in the same holothurian host during which they produce sounds. Another characteristic of these fish lies in their abilities to produce sounds. The objective of this study was dual: (1) to seek if there was a sexual difference in the sounds produced by C. boraborensis; (2) to seek if there was a difference in the sound emissions between heterospecific and conspecific encounters. In each trial, sounds were only recorded when one individual entered the sea cucumber that was already occupied. In encounters, sounds were structured in regular pulse emissions whose pulse lengths and periods allowed to significantly distinguish each species, as well as both sexes in C. boraborensis. In the latter species, results show for the first time that temporal features of the emitted sounds can have a functional importance in sex identification. In heterospecific encounters, sounds were reduced 68% of the time to a single pulse emission and there was a modification in the pulse length of each species: it shortens in C. homei and it lengthens in C. boraborensis. It highlights that both carapids are able to adapt their sounds to the facing species. Because a modification of the sound appears to be done at the first emission, it is supposed that recognition precedes the sound emission. PMID- 16254914 TI - Influence of body reserves and eye opacity on foraging behaviours of tiger snakes. AB - Physiological states such as reproductive stage, parasite load, body condition, and environmental conditions, influence behaviours in complex ways. According to the Clark's asset-protection principle (Clark, 1994. Behav Ecol 5:159-170), individuals in good body condition take fewer predation risks than conspecifics in lesser condition. In many ectotherms, foraging, digestion, and moulting require an elevation of the metabolism achieved through intensive basking, thereby increasing the risk of being detected by predators. Using four experimental groups of snakes, we showed that two independent physiological states, (1) the presence of prey in the stomach and (2) eye opacity associated with moulting, increase predator vulnerability. In a parallel experiment, two groups of snakes were maintained on contrasting diets (high food intake vs. low food intake) for 6 months. Apart from sloughing periods, most individuals (89%) accepted their meal irrespective of food treatment. Consequently, well-fed snakes exhibited higher body condition (one third physiological state) relative to less fed individuals. During sloughing events, however, well-fed individuals often refused (78%) to eat while the less-fed individuals readily accepted (86%) their meal despite eye opacity. Individuals with less body reserves accepted the cumulative risks owing to eye opacity and stomach fullness. By contrast, well-fed snakes remained more "prudent" (i.e., exhibited behaviours that depended on their internal state). Our results show that snakes can adjust their foraging "decision" by combining different physiological informations. PMID- 16254915 TI - Continuous male presence required for fertilization in captive painted dragons, Ctenophorus pictus. AB - Understanding proximate determinants of reproductive events is an important issue for many biologists. In the present paper, we show that follicular recruitment and fertilization are dependent on male stimuli in long-term captive female painted dragon lizards, Ctenophorus pictus. Hibernated females kept separated from males did not initiate reproduction, despite visual contact with displaying males and repeated copulation attempts. Follicular stimulating hormone stimulated ovarian growth and female receptivity, but did not result in successful fertilization of eggs, despite repeated copulations. When housed continuously together with males, however, follicle recruitment and fertilization were achieved. Thus, not only ovarian growth, but also fertilization, is dependent on male stimuli. PMID- 16254916 TI - Nuclear transfer of M-phase ferret fibroblasts synchronized with the microtubule inhibitor demecolcine. AB - The development of reconstructed embryos following nuclear transfer (NT) appears to be dependent upon a variety of factors, including cell cycle synchronization between the donor nucleus and recipient oocyte. Here we use the microtubule inhibitor, demecolcine, to synchronize ferret fibroblasts in metaphase (M-phase) in order to match their cell cycle position with that of the recipient oocyte at the time of NT. The fibroblasts were obtained from 28-day fetuses and cultured for 1-30 days prior to NT. Fibroblast cultures were treated with 0.05 microg/ml of demecolcine for 3 hr or overnight (14-16 hr) after various times in culture to determine the optimal conditions for M-phase synchronization. The percentage of G2/M-phase cells in demecolcine-treated cultures was significantly greater than that found in untreated cultures (P<0.05). Optimally synchronized M-phase fibroblasts were collected by mitotic shake-off and evaluated for their effectiveness in NT. M-phase somatic cell-derived NT embryos reconstituted by electrofusion or microinjection underwent implantation and formed fetuses at similar rates (5.4% vs. 3.4%, and 1.8% vs. 1.2%, respectively); however, no NT embryos developed to term. In summary, these data demonstrate two important points. First, demecolcine treatment effectively synchronizes ferret fibroblasts in M-phase of the cell cycle; and second, these somatic cells are capable of driving embryo development following NT. Our results should facilitate the development of cloned ferrets as an animal model for human lung disease such as influenza and cystic fibrosis. PMID- 16254917 TI - Tail regeneration and ependymal outgrowth in the adult newt, Notophthalmus viridescens, are adversely affected by experimentally produced ischemia. AB - Spinal axons of the adult newt will regenerate when the spinal cord is severed or when the tail is amputated. Ischemia and associated hypoxia have been correlated with poor central nervous system regeneration in mammals. To test the effects of ischemia on newt spinal cord regeneration, the spinal cord and major blood vessels of the newt tail were severed 2 cm caudal to the cloaca as a primary injury. This primary injury severely reduced circulation in the caudal direction for 7 days; by day 8, circulation was largely restored. After various periods of time after primary injury, tails were amputated 1 cm caudal to the primary injury (in the area of ischemia) and tested for regeneration. If the tail was amputated within 5 days of the primary injury, regeneration did not occur. If amputation was 7 days or longer after the primary injury, a regenerative response occurred. Histology showed that in the non-regenerating tails the spinal cord and associated ependyma, known to be important to tail regeneration, had degenerated in the rostral direction. Such degeneration was prevented when tails were first amputated and allowed to form blastemas before the primary injury. The data indicate that the first 5-7 days of blastema formation are particularly sensitive to compromised blood flow (ischemia/hypoxia). It follows that mechanisms must be present in the adult newt to reduce ischemia to a minimum and thus allow ependymal outgrowth and tail regeneration. PMID- 16254918 TI - The interplay of increased urea synthesis and reduced ammonia production in the African lungfish Protopterus aethiopicus during 46 days of aestivation in a mucus cocoon. AB - This study was undertaken to test the hypothesis that the rate of urea synthesis in Protopterus aethiopicus was up-regulated to detoxify ammonia during the initial phase of aestivation in air (day 1-day 12), and that a profound suppression of ammonia production occurred at a later phase of aestivation (day 35-day 46) which eliminated the need to sustain the increased rate of urea synthesis. Fasting apparently led to a greater rate of nitrogenous waste excretion in P. aethiopicus in water, which is an indication of increases in production of endogenous ammonia and urea probably as a result of increased proteolysis and amino acid catabolism for energy production. However, 46 days of fasting had no significant effects on the ammonia or urea contents in the muscle, liver, plasma and brain. In contrast, there were significant decreases in the muscle ammonia content in fish after 12, 34 or 46 days of aestivation in air when compared with fish fasting in water. Ammonia was apparently detoxified to urea because urea contents in the muscle, liver, plasma and brain of P. aethiopicus aestivated for 12, 34 or 46 days were significantly greater than the corresponding fasting control; the greatest increases in urea contents occurred during the initial 12 days. There were also significant increases in activities of some of the hepatic ornithine-urea cycle enzymes from fish aestivated for 12 or 46 days. Therefore, contrary to a previous report on P. aethiopicus, our results demonstrated an increase in the estimated rate of urea synthesis (2.8 fold greater than the day 0 fish) in this lungfish during the initial 12 days of aestivation. However, the estimated rate of urea synthesis decreased significantly during the next 34 days. Between day 35 and day 46 (12 days), urea synthesis apparently decreased to 42% of the day 0 control value, and this is the first report of such a phenomenon in African lungfish undergoing aestivation. On the other hand, the estimated rate of ammonia production in P. aethiopicus increased slightly (14.7%) during the initial 12 days of aestivation as compared with that in the day 0 fish. By contrast, the estimated rate of ammonia production decreased by 84% during the final 12 days of aestivation (day 35-day 46) compared with the day 0 value. Therefore, it can be concluded that P. aethiopicus depended mainly on increased urea synthesis to ameliorate ammonia toxicity during the initial phase of aestivation, but during prolonged aestivation, it suppressed ammonia production profoundly, eliminating the need to increase urea synthesis which is energy-intensive. PMID- 16254919 TI - Effects of pharmacological treatment and photoinactivation on the directional responses of an insect neuron. AB - Soma-ipsilateral branches of the large segmental omega neuron of the phaneropterid bush cricket Ancistrura nigrovittata have smooth endings, which extend through most of the auditory neuropile. Correspondingly, it shows a broad frequency tuning. Large excitatory postsynaptic potentials (EPSPs) and inhibitory postsynaptic potentials (IPSPs) are observed when recording from soma-ipsilateral branches. Stimulation from the soma-ipsilateral side leads to a strong excitation. Soma-contralateral branches have a strong, beaded appearance. IPSPs, which seem to be of soma-contralateral origin, can be recorded from these branches. Stimulation from the soma-contralateral side leads to a strong inhibition of the omega neuron. Soma-contralateral stimulation must be 30-40 dB more intense than soma-ipsilateral stimulation to evoke similar spike numbers in the omega neuron. The side-to-side difference is reduced to 10-15 dB after cutting the input from the soma-contralateral leg (tympanic nerve). The thresholds for eliciting IPSPs by soma-contralateral stimulation correspond roughly to excitatory thresholds of the mirror-image omega with the same stimuli. Pharmacological treatment with picrotoxin (PTX) or photoinactivation of the Lucifer Yellow filled mirror-image omega neuron reduces contralateral inhibition considerably and eliminates all visible IPSPs. Nevertheless, an additional contralateral inhibition survives both procedures and is only eliminated after cutting the soma-contralateral tympanic nerve. These results demonstrate that the mirror-image partners of the omega neuron mutually inhibit each other in bush crickets--as in crickets. This mutual inhibition is PTX-sensitive. At least one additional element exerts contralateral PTX-insensitive inhibition on the omega neuron. PMID- 16254920 TI - Development of gene transfer technology for black tiger shrimp, Penaeus monodon. AB - An effective foreign gene transfer method for shrimp would have several potential uses in the shrimp culture industry, such as in preventing infectious diseases. We evaluated two gene transfer methods and used black tiger shrimp, Penaeus monodon, as a model target species. For a promoter, we used the 1,592-bp promoter region of the EF-1alpha gene, a house-keeping gene, of kuruma shrimp Marsupenaeus japonicus. The promoter region was linked to either the gene for green fluorescence protein (GFP) or the gene for chloramphenicol acetyl transferase (CAT). The fusion genes were designated pJEF-GFP and pJEF-CAT, respectively. The pJEF-GFP gene was introduced into fertilized eggs of black tiger shrimp by microinjection and particle gun bombardment. The survival rate of the microinjected eggs was 17.6%, and 1.0% of the treated embryos were found to be GFP-positive. However, the GFP-positive embryos were damaged and embryogenesis did not progress. The survival rate of the particle-bombarded eggs was 60.6%, and 0.42% of the treated embryos were found to be GFP-positive. Ubiquitous GFP expression was observed from 8 hr post-fertilization and these embryos developed and hatched normally. The pJEF-CAT gene was introduced into fertilized eggs of black tiger shrimp using the optimized conditions of the particle gun bombardment. CAT activity was observed from 1 to 7 days post-fertilization, with the highest activities being observed at 5 and 7 days post-hatching. PMID- 16254921 TI - Is sexual dimorphism affected by the combined action of prenatal stress and sex ratio? AB - During embryonic development, offspring are exposed to hormones of both maternal and sibling origin. Maternal stress increases offspring exposure to corticosterone, and, in polytocous animals, the sex ratio or intrauterine position can influence the levels of androgens and estrogens experienced by the offspring. Such hormone exposure has the potential to influence many important morphological and behavioural aspects of offspring, in particular sexually dimorphic traits. Although well known in rodents, the impact of prenatal hormone exposure in other vertebrates is poorly documented. We experimentally investigated the relationship between maternal stress, population density, sex ratio (a surrogate for the degree of exposure to steroids produced by siblings), and sexual dimorphism in a viviparous lizard, Lacerta vivipara. Our results show that prenatal sex ratios have consequences for sexually dimorphic morphology (ventral scale count) in both sexes, but with no effect of maternal stress or any interaction between the two. Embryonic steroid exposure can potentially be an important factor in generating individual variation in natural populations of viviparous animals. PMID- 16254922 TI - Follicular fluid steroid levels and subsequent sex of bovine embryos. AB - On the basis of evidence suggesting a maternal involvement in the determination of the sex of the offspring, we took ova at the point of ovulation from crossbred heifers, fertilised them, and established the sex of the embryos. At the same time we took individual-matched samples of follicular fluid from each follicle of origin, and measured the levels of testosterone and oestradiol, blind to the sex of the embryo. We found no effect of oestradiol on sex in either primary or subordinate follicles. But bovine ova from subordinate follicles that had follicular fluid with a high concentration of testosterone (in vivo) were later more likely to be fertilised by a Y-chromosome-bearing spermatozoon (in vitro). These, along with similar results from other researchers, suggest that further study of the relationship between mammalian follicular hormones at the time of conception and subsequent sex of offspring, may help resolve some of the problems associated with theories of adaptive control of the sex ratio in mammals. PMID- 16254923 TI - Increases in urea synthesis and the ornithine-urea cycle capacity in the giant African snail, Achatina fulica, during fasting or aestivation, or after the injection with ammonium chloride. AB - The objectives of this study are to determine whether a full complement of ornithine-urea cycle (OUC) enzymes is present in the hepatopancreas of the giant African snail Achatina fulica, and to investigate whether the rate of urea synthesis and the OUC capacity can be up-regulated during 23 days of fasting or aestivation, or 24 hr post-injection with NH(4)Cl (10 micromol g(-1) snail) into the foot muscle. A. fulica is ureotelic and a full complement of OUC enzymes, including carbamoyl phosphate synthetase III (CPS III), was detected from its hepatopancreas. There were significant increases in the excretion of NH(4)(+), NH(3) and urea in fasting A. fulica. Fasting had no significant effect on the tissue ammonia contents, but led to a progressive accumulation of urea, which was associated with an 18-fold increase in the rate of urea synthesis. Because fasting took place in the presence of water and because there was no change in water contents in the foot muscle and hepatopancreas, it can be concluded that the function of urea accumulation in fasting A. fulica was unrelated to water retention. Aestivation in arid conditions led to a non-progressive accumulation of urea in A. fulica. During the first 4 days and the last 3 days of the 23-day aestivation period, experimental snails exhibited significantly greater rates of urea synthesis compared with fasted snails. These increases were associated with significant increases in activities of various OUC enzymes, except CPS III, in the hepatopancreas. However, the overall urea accumulation in snails aestivated and snails fasted for 23 days were comparable. Therefore, the classical hypothesis that urea accumulation occurred to prevent water loss through evaporation during aestivation in terrestrial pulmonates may not be valid. Surprisingly, there were no accumulations of ammonia in the foot muscle and hepatopancreas of A. fulica 12 or 24 hr after NH(4)Cl was injected into the foot muscle. In contrast, the urea content in the foot muscle of A. fulica increased 4.5- and 33-fold at hour 12 and hour 24, respectively, and the respective increases in the hepatopancreas were 4.9- and 32-fold. The exogenous ammonia injected into A. fulica was apparently detoxified completely to urea. The urea synthesis rate increased 148-fold within the 24-hr experimental period, which could be the greatest increase known among animals. Simultaneously, there were significant increases in activities of glutamine synthetase (2.5-fold), CPS III (3.1-fold), ornithine transcarbamoylase (2.3-fold), argininosuccinate synthetase+lyase (13.6-fold) and arginase (3.5-fold) in the hepatopancreas 12 hr after the injection of NH(4)Cl. Taken altogether, our results support the view that the primary function of urea synthesis through the OUC in A. fulica is to defend against ammonia toxicity, but suggest that urea may have more than an excretory role in terrestrial pulmonates capable of aestivation. PMID- 16254924 TI - Proteomic analysis of sera from hepatocellular carcinoma patients after radiofrequency ablation treatment. AB - Comparative proteomic analysis was used to search for characteristic alterations in the sera of hepatocellular carcinoma (HCC) patients who had undergone curative radiofrequency ablation treatment. Serum samples collected from eight patients before and after treatment were subjected to 2-DE. Eighty-eight protein spots differentially expressed with the treatment were selected by clustering analysis, and the proteins were identified by MS based on MALDI-TOF/TOF analysis and public database searches. The statistical analysis suggested that four proteins decreased after treatment (pro-apolipoprotein, alpha2-HS glycoprotein, apolipoprotein A-IV precursor, and PRO1708/PRO2044, which is the carboxy terminal fragment of albumin) and that seven proteins were increased after treatment, including leucine-rich alpha2-glycoprotein and alpha1-antitrypsin. These data facilitate the identification of differentially expressed proteins that are involved in HCC carcinogenesis and provide candidate biomarkers for the development of diagnostic and therapeutic tools. PMID- 16254927 TI - High throughput production of mouse monoclonal antibodies using antigen microarrays. AB - Recent advances in proteomics research underscore the increasing need for high affinity monoclonal antibodies, which are still generated with lengthy, low throughput antibody production techniques. Here we present a semi-automated, high throughput method of hybridoma generation and identification. Monoclonal antibodies were raised to different targets in single batch runs of 6-10 wk using multiplexed immunisations, automated fusion and cell-culture, and a novel antigen coated microarray-screening assay. In a large-scale experiment, where eight mice were immunized with ten antigens each, we generated monoclonal antibodies against 68 of the targets (85%), within 6 wk of the primary immunization. PMID- 16254928 TI - Improved peak detection and quantification of mass spectrometry data acquired from surface-enhanced laser desorption and ionization by denoising spectra with the undecimated discrete wavelet transform. AB - Mass spectrometry is being used to find disease-related patterns in mixtures of proteins derived from biological fluids. Questions have been raised about the reproducibility and reliability of peak quantifications using this technology. We collected nipple aspirate fluid from breast cancer patients and healthy women, pooled them into a quality control sample, and produced 24 replicate SELDI spectra. We developed a novel algorithm to process the spectra, denoising with the undecimated discrete wavelet transform (UDWT), and evaluated it for consistency and reproducibility. UDWT efficiently decomposes spectra into noise and signal. The noise is consistent and uncorrelated. Baseline correction produces isolated peak clusters separated by flat regions. Our method reproducibly detects more peaks than the method implemented in Ciphergen software. After normalization and log transformation, the mean coefficient of variation of peak heights is 10.6%. Our method to process spectra provides improvements over existing methods. Denoising using the UDWT appears to be an important step toward obtaining results that are more accurate. It improves the reproducibility of quantifications and supplies tools for investigation of the variations in the technology more carefully. Further study will be required, because we do not have a gold standard providing an objective assessment of which peaks are present in the samples. PMID- 16254929 TI - Global protein expression pattern of Bradyrhizobium japonicum bacteroids: a prelude to functional proteomics. AB - As a prelude to using functional proteomics towards understanding the process of symbiotic nitrogen fixation between the legume soybean and the soil bacteria Bradyrhizobium japonicum, we examined the total protein expression pattern of the nodule bacteria, often referred to as bacteroids. A partial proteome map was constructed by separating the total bacteroid proteins using high-resolution 2 DE. Of the several hundred protein spots analyzed using PMF, 180 spots were tentatively identified by searching the available database for B. japonicum, (http://www.kazusa.or.jp/index.html). The data showed that the bacteroid expressed a dominant and elaborate protein network for nitrogen and carbon metabolism, which is closely dependent on the plant supplied metabolites, and seems aptly supported by a selective group of bacteroid transporter proteins. However, they seem to lack a defined fatty acid and nucleic acid metabolism. Interestingly, the proteins related to protein synthesis, scaffolding and degradation were among the most predominant spots of the bacteroid proteome. In addition, several proteins, which showed fairly good expression, were identified to be involved with cellular detoxification, stress regulation and signaling communication components. This preliminary proteomic data matches very well with several biochemical and genetic reports, and clearly shows the inter-connection between several metabolic pathways that meet the needs of the bacteroid. It is expected that in the future this will allow us to develop testable hypotheses about the roles of several of these proteins in context to the metabolic pathway connections and metabolite fluxes. PMID- 16254930 TI - Identification of Arabidopsis salt and osmotic stress responsive proteins using two-dimensional difference gel electrophoresis and mass spectrometry. AB - Arabidopsis thaliana cell suspension cultures have been used to investigate the effects of salinity and hyperosmotic stress on plant cellular proteins. We show that 200 mM NaCl and 400 mM sorbitol treatments induce extracellular medium acidification in Arabidopsis cell cultures, a typical response of plant cells to salt and hyperosmotic stress. Using (35)S-labelled amino acids, we demonstrated that NaCl causes a transient suppression of de novo protein synthesis, from which the cells recover within 4 h. Changes in the abundance of cellular proteins 6 h post NaCl and sorbitol treatments were analysed by 2-DE. Of a total of 2,949 protein spots detected on the gels, 266 showed significant changes in abundance across five independent experiments. Using MALDI-TOF MS, we identified 75 salt and sorbitol responsive spots. These fall into 10 functional categories that include H(+) transporting ATPases, signal transduction related proteins, transcription/translation related proteins, detoxifying enzymes, amino acid and purine biosynthesis related proteins, proteolytic enzymes, heat-shock proteins, carbohydrate metabolism-associated proteins and proteins with no known biological functions. PMID- 16254931 TI - Development of a novel chemical probe for the selective enrichment of phosphorylated serine- and threonine-containing peptides. AB - Gaining insight into phosphoproteomes is of the utmost importance for understanding regulation processes such as signal transduction and cellular differentiation. While the identification of phosphotyrosine-containing amino acid sequences in peptides and proteins is now becoming possible, mainly because of the availability of high-affinity antibodies, no general and robust methodology allowing the selective enrichment and analysis of serine- and threonine-phosphorylated proteins and peptides is presently available. The method presented here involves chemical modification of phosphorylated serine or threonine residues and their subsequent derivatization with the aid of a multifunctional probe molecule. The designed probe contains four parts: a reactive group that is used to bind specifically to the modified phosphopeptide, an optional part in which heavy isotopes can be incorporated, an acid-labile linker, and an affinity tag for the selective enrichment of modified phosphopeptides from complex mixtures. The acid-cleavable linker allows full recovery from the affinity-purified material and removal of the affinity tag prior to MS analysis. The preparation of a representative probe molecule containing a biotin affinity tag and its applicability in phosphoproteome analysis is shown in a number of well-defined model systems of increasing degrees of complexity. Amounts of phosphopeptide as low as 1 nmol can be modified and enriched from a mixture of peptides. During the development of the beta elimination/nucleophilic addition protocol, special attention was paid to the different experimental parameters that might affect the chemical-modification steps carried out on phosphorylated residues. PMID- 16254932 TI - Quantum-dot aptamer beacons for the detection of proteins. PMID- 16254933 TI - Synthesis of [all]-S,S-dioxide oligothiophenes using HOF.CH3CN. PMID- 16254935 TI - S-heterocyclic carbene with a disilane backbone. PMID- 16254934 TI - Conversion of a carboxylesterase into a triacylglycerol lipase by a random mutation. PMID- 16254936 TI - Ruthenium-catalyzed asymmetric propargylic substitution reactions of propargylic alcohols with acetone. PMID- 16254937 TI - Solid film versus solution-phase charge-recombination dynamics of exTTF-bridge C60 dyads. AB - The charge-recombination dynamics of two exTTF-C60 dyads (exTTF = 9,10-bis(1,3 dithiol-2-ylidene)-9,10-dihydroanthracene), observed after photoinduced charge separation, are compared in solution and in the solid state. The dyads differ only in the degree of conjugation of the bridge between the donor (exTTF) and the acceptor (C60) moieties. In solution, photoexcitation of the nonconjugated dyad C60-BN-exTTF (1) (BN = 1,1'-binaphthyl) shows slower charge-recombination dynamics compared with the conjugated dyad C60-TVB-exTTF (2) (TVB = bisthienylvinylenebenzene) (lifetimes of 24 and 0.6 micros, respectively), consistent with the expected stronger electronic coupling in the conjugated dyad. However, in solid films, the dynamics are remarkably different, with dyad 2 showing slower recombination dynamics than 1. For dyad 1, recombination dynamics for the solid films are observed to be tenfold faster than in solution, with this acceleration attributed to enhanced electronic coupling between the geminate radical pair in the solid film. In contrast, for dyad 2, the recombination dynamics in the solid film exhibit a lifetime of 7 micros, tenfold slower than that observed for this dyad in solution. These slow recombination dynamics are assigned to the dissociation of the initially formed geminate radical pair to free carriers. Subsequent trapping of the free carriers at film defects results in the observed slow recombination dynamics. It is thus apparent that consideration of solution-phase recombination data is of only limited value in predicting the solid-film behaviour. These results are discussed with reference to the development of organic solar cells based upon molecular donor-acceptor structures. PMID- 16254938 TI - Columnar and smectic liquid crystals based on crown ethers. AB - Unsymmetrical benzo[15]crown-5 ethers 5 with one lateral ortho-terphenyl unit bearing alkoxy side chains of varying chain lengths (C5-C14) were prepared from 3,4-dialkoxyphenylbromides 2. Complexation with metal salts MX (M = Na, Cs) afforded the corresponding derivatives MX5. The uncomplexed crown ethers 5 h and 5 i, with dodecyloxy and tetradecyloxy side chains, respectively, exhibit liquid crystalline properties. In the series of complexed crown ethers, liquid crystal properties appeared as early as NaI5 f with C9H19 side chains. Whereas the uncomplexed 5 h,i form smectic mesophases, the complexed NaI5 g and NaI5 h exhibit textures typical of columnar mesophases. These results were supported by X-ray diffraction measurements (WAXS, SAXS), which revealed smectic (5 h,i), rectangular columnar (NaI5 g), and hexagonal columnar (NaI5 h) mesophases. As the liquid crystalline phase might retain packing features of the solid-state structure, single-crystal X-ray analyses were also performed for some of the uncomplexed and complexed crown ether derivatives. The complex NaI(3)5 a displays a sandwich-type structure, with the crown ether cores mutually antiperiplanar and maintaining an almost perfect crown conformation. In contrast, non-mesogenic uncomplexed crown ether 5 b displays a layer-type ordering in the solid phase. PMID- 16254939 TI - Electron-rich tetrathiafulvalene-triarylamine conjugates: synthesis and redox properties. AB - By combining tetrathiafulvalenes (TTFs) and triarylamines, four TTF-triarylamine conjugates bridged by an annulated pyrrole ring were designed and synthesized by an N-arylation reaction. Electrochemical and photophysical investigations suggest that these novel conjugates possess very strong electron-donating ability with very high HOMO energy levels of around -4.70 eV; the HOMOs are mainly located on the TTF moiety. We observed significant electronic coupling between the TTF moieties and the triarylamine groups. However, no evidence for such electronic communication between end-capping TTF units (conjugates 5 and 7) or between two terminal triarylamine groups (conjugate 9) could be found. Differential scanning calorimetry (DSC) measurements together with PM3-optimized geometries suggest that conjugates 5 and 7, which adopt three-dimensional propeller-shaped structures, may easily pack and crystallize in the solid state because of the large rigid planar blades consisting of TTF and one of the phenyl rings of the triarylamine moiety. However, conjugate 9, with two bulky end-capping triarylamine groups, forms an amorphous material with a glass transition at 74.5 degrees C. PMID- 16254940 TI - Break on through to the other side-biophysics and cell biology shed light on cell penetrating peptides. AB - Cell-penetrating peptides (CPPs) have become widely used vectors for the cellular import of molecules in basic and applied biomedical research. Despite the broad acceptance of these molecules as molecular carriers, the details of the mode of cellular internalization and membrane permeation remain elusive. Within the last two years endocytosis has been demonstrated to be a route of uptake shared by several CPPs. These findings had a significant impact on CPP research. State-of the-art cell biology is now required to advance the understanding of the intracellular fate of the CPP and cargo molecules. Owing to their presumed ability to cross lipid bilayers, CPPs also represent highly interesting objects of biophysical research. Numerous studies have investigated structure-activity relationships of CPPs with respect to their ability to bind to a lipid bilayer or to cross this barrier. Endocytosis route only relocates the membrane permeation from the cell surface to endocytic compartments. Therefore, biophysical experiments are key to a mechanistic molecular understanding of the cellular uptake of CPPs. However, biophysical investigations have to consider the molecular environment encountered by a peptide inside and outside a cell. In this contribution we will review biophysical and cell-biology data obtained for several prominent CPPs. Furthermore, we will summarize recent findings on the cell-penetrating characteristics of antimicrobial peptides and the antimicrobial properties of CPPs. Peptides of both groups have overlapping characteristics. Therefore, both fields may greatly benefit from each other. The review will conclude with a perspective of how biophysics and cell biology may synergize even more efficiently in the future. PMID- 16254941 TI - Design of a hairpin polyamide, ZT65B, for targeting the inverted CCAAT box (ICB) site in the multidrug resistant (MDR1) gene. AB - A novel hairpin polyamide, ZT65B, containing a 3-methylpicolinate moiety was designed to target the inverted CCAAT box (ICB) of the human multidrug resistance 1 gene (MDR1) promoter. Binding of nuclear factor-Y (NF-Y) to the ICB site upregulates MDR1 gene expression and is, therefore, a good target for anticancer therapeutic agents. However, it is important to distinguish amongst different promoter ICB sites so that only specific genes will be affected. All ICB sites have the same sequence but they differ in the sequence of the flanking base pairs, which can be exploited in the design of sequence-specific polyamides. To test this hypothesis, ten ICB-containing DNA hairpins were designed with different flanking base pairs; the sequences ICBa and ICBb were similar to the 3' ICB site of MDR1 (TGGCT). Thermal-denaturation studies showed that ZT65B effectively targeted ICBa and ICBb (DeltaTM=6.5 and 7.0 degrees C) in preference to the other DNA hairpins (<3.5 degrees C), with the exception of ICBc (5.0 degrees C). DNase I-footprinting assays were carried out with the topoisomerase IIalpha-promoter sequence, which contains five ICB sites; of these, ICB1 and ICB5 are similar to the ICB site of MDR1. ZT65B was found to selectively bind ICB1 and ICB5; footprints were not observed with ICB2, ICB3, or ICB4. A strong, positive induced ligand band at 325 nm in CD studies confirmed that ZT65B binds in the DNA minor groove. The selectivity of ZT65B binding to hairpins that contained the MDR1 ICB site compared to one that did not (ICBd) was confirmed by surface plasmon studies, and equilibrium constants of 5x10(6)-1x10(7) and 4.6x10(5) M-1 were obtained with ICB1, ICB5,and ICB2 respectively. ZT65B and the previously published JH37 (J. A. Henry, et al. Biochemistry 2004, 43, 12 249-12 257) serve as prototypes for the design of novel polyamides. These can be used to specifically target the subset of ubiquitous gene elements known as ICBs, and thereby affect the expression of one or a few proteins. PMID- 16254942 TI - Ligand binding transmits conformational changes across the membrane-spanning region to the intracellular side of the 5-HT3 serotonin receptor. PMID- 16254944 TI - B(0) and B(1)-insensitive uniform T(1)-weighting for quantitative, first-pass myocardial perfusion magnetic resonance imaging. AB - Myocardial perfusion can be estimated, in principle, from first-pass MR images by converting the T(1)-weighted signal-time curves to contrast agent concentration time curves. Typically, T(1) weighting is achieved by saturating the magnetization with a nonselective radiofrequency (RF) pulse prior to the imaging sequence. The accuracy of the perfusion estimate derived from the single-point T(1)-weighted signal depends on the initial residual longitudinal magnetization (RLM) produced by the saturation pulse. In this study we demonstrate that single shot, echo-planar imaging can be used to show initial RLM resulting from incomplete saturation due to static magnetic field and RF field inhomogeneities in the heart at 1.5 T. Three saturation pulses, single, composite simple, and composite B(1)-insensitive rotation (BIR-4) were evaluated in phantom and cardiac experiments. The RLM image was calculated by normalizing the saturated image by a proton-density-weighted image. Mean RLM produced by the three saturation pulses was significantly different in noncontrast cardiac imaging (RLM(single) = 0.108 +/- 0.078; RLM(composite) = 0.051 +/- 0.052; RLM(BIR-4) = 0.011 +/- 0.009; P < 0.001; n = 20). Using a BIR-4 pulse to perform saturation of magnetization seems promising for improving the effectiveness and uniformity of T(1) weighting for first-pass perfusion imaging. PMID- 16254945 TI - HR-MAS of cells: A "cellular water shift" due to water-protein interactions? AB - Under HR-MAS conditions, cells are subjected to high centrifugal forces that may cause irreversible cell damage. First, conditions have been defined to monitor and keep to a minimum unwanted effects in HR-MAS spectra arising from the loss of cell integrity. Then, the HR-MAS spectra of reasonably intact cells have been analyzed. Cell suspensions subjected to MAS rates as low as 1 kHz split into a two-compartment system that is composed of a cell-rich phase (H(2)O(i)) and a cell-free phase (H(2)O(o)). Each of these phases is characterized by its own water (1)H-NMR signal. Transport of water molecules between the cell-rich and cell-free compartments is limited by the very low contact area between the two compartments, and water exchange dynamics consequently fall into the slow exchange limit on the NMR timescale. Since the exchange between the two water populations is "frozen," the separation between the H(2)O(o) and H(2)O(i) water signals (Deltanu(water)) detected in an HR-MAS experiment is not affected by chemical exchange but reflects only chemical differences in the two environments. Different cell lines show a different Deltanu(water), leading to the concept of "cellular water shift." This shift roughly correlates with the cellular protein content, supporting the view that the most important determinant of the cellular water shift is the interaction between water and proteins in the intracellular compartment. PMID- 16254946 TI - High-resolution DTI with 2D interleaved multislice reduced FOV single-shot diffusion-weighted EPI (2D ss-rFOV-DWEPI). AB - Diffusion tensor MRI (DTI), using single-shot 2D diffusion weighted-EPI (2D ss DWEPI), is limited to intracranial (i.c.) applications far from the sinuses and bony structures, due to the severe geometric distortions caused by significant magnetic field inhomogeneities at or near the tissue-air or tissue-bone interfaces. Reducing these distortions in single-shot EPI by shortening the readout period generally requires a reduced field of view (and the potential of wraparound artifact) in the phase-encoding direction and/or reduced spatial resolution. To resolve the problem, a novel 2D reduced FOV single-shot diffusion weighted EPI (2D ss-rFOV-DWEPI) pulse sequence applicable for high resolution diffusion-weighted MRI of local anatomic regions, such as brainstem, cervical spinal cord, and optic nerve, has been developed. In the proposed technique, time efficient interleaved acquisition of multiple slices with a limited FOV was achieved by applying an even number of refocusing 180 degrees pulses with the slice-selection gradient applied in the phase-encoding direction. The two refocusing pulses used for each slice acquisition were separated by a short time interval (typically less than 45 ms) required for the 2D EPI echotrain acquisition. The new technique can be useful for high resolution DTI of various anatomies, such as localized brain structures, cervical spinal cord, optic nerve, heart, or other extra-cerebral organ, where conventional 2D ss-DWEPI is limited in usage due to the severity of image distortions. PMID- 16254947 TI - The effect of ergonomic interventions in healthcare facilities on musculoskeletal disorders. AB - BACKGROUND: The high incidence of musculoskeletal disorders (MSDs) among healthcare workers suggests that the introduction of ergonomic interventions could be beneficial. While laboratory studies have clearly documented the efficacy of ergonomic devices, few studies have examined their effectiveness in the healthcare workplace. METHODS: This study evaluated a statewide program that provided ergonomic consultation and financial support for purchasing ergonomic devices, which aid in patient handling and lifting. Changes in MSD rates between baseline (1 year pre-intervention) and post-intervention (up to 2 years) periods were examined in 100 work units in 86 healthcare facilities. RESULTS: The median MSD rate decreased from 12.32 to 6.64 per 200,000 employee-hours, a decrease greater than the secular trend for the study period (1999-2003). CONCLUSIONS: This study suggests that ergonomic consultation and financial support for purchasing ergonomic equipment can be an effective intervention to reduce MSDs among healthcare workers. PMID- 16254948 TI - Machine safety evaluation in small metal working facilities: an evaluation of inter-rater reliability in the quantification of machine-related hazards. AB - BACKGROUND: Each year there are an estimated 4.2-6.7 amputations per 10,000 workers in the metal fabrication trades in the United States. The Minnesota Machine Guarding Study evaluates the effectiveness of a peer-based technical and educational intervention designed to reduce exposure to amputation hazards among workers in small machining/metal working shops. The study reported here involved the development and evaluation of methods for measuring machine safety, which will be used in the intervention study. METHODS: Using OSHA regulations, ANSI standards, and industry best practices, we developed 23 machine-safety scorecards. The safety scores were dependent on the presence or absence of guards, other safety devices and implements, and the presence or absence of acceptable administrative programs. Inter-rater reliability was assessed for the evaluation of eight types of commonly used metal fabrication machines. RESULTS: Of the 23 most common types of machines, there were a sufficient number of machines to evaluate inter-rater reliability for eight types. Three raters in four shops assessed fifty-nine machines. Overall, the kappa statistic ranged from 0.57 to 0.84, indicating good to very good concordance between raters. In general, machines did not fare well with regard to compliance with current standards. DISCUSSION: The ability to assess machine-related hazards is important in industries where it is difficult to identify and count injuries in a timely fashion. Machinists and safety professionals may use this scoring system as a means of reproducible assessment of machine safety. PMID- 16254949 TI - Surveillance of hearing loss among older construction and trade workers at Department of Energy nuclear sites. AB - BACKGROUND: Medical screening programs at three Departments of Energy (DOE) nuclear weapons facilities (Hanford Nuclear Reservation, Oak Ridge, and the Savannah River Site) have included audiometric testing since approximately 1996. This report summarizes hearing evaluations through March 31, 2003. METHODS: Occupational examinations included a medical history, limited physical examination, and tests for medical effects from specific hazards, including audiometric testing. Hearing thresholds by frequency for DOE workers were compared to age-standardized thresholds among an external comparison population of industrial workers with noise exposures <80 dBA. Multivariate analyses were used to explore the risk of hearing impairment by duration of construction trade work and self-reported noise exposure, while controlling for potential confounders such as age, race, sex, smoking, elevated serum cholesterol, hypertension, solvent exposures, and recreational noise exposures. RESULTS: Hearing thresholds among DOE workers were much higher than observed in a comparison population of industrial workers with low noise exposures. Overall, 59.7% of workers examined were found to have material hearing impairment by NIOSH criteria. Age, duration of construction work, smoking, and self-reported noise exposure increased the risk of hearing loss. The risk of material hearing impairment was significantly elevated for construction trade workers compared to the external comparison population (odds-ratio = 1.6, 95% CI = 1.3-2.1) and increased with the duration of trade work. CONCLUSIONS: These medical screening programs confirm worker concerns about risks for hearing loss and the need for hearing conservation programs for construction workers, with emphasis on the prevention of noise exposures. PMID- 16254950 TI - Pulmonary abnormalities associated with occupational exposures at the Savannah River Site. AB - BACKGROUND: A program of medical evaluation for former Savannah River Site (SRS) workers at health effects due to exposures to hazardous or radioactive agents was conducted. METHODS: This study includes data from 1,368 participants aged 45 years or older who were assessed regarding work-history and exposures to industrial agents. According to the standard industrial classification (SIC), participants were employed in five of the SIC divisions. Based on the International Labour Office Classification of Radiographs, two categories of pleural and parenchymal abnormalities were evaluated by a single radiologist. The SRS results were compared with the second national health and nutrition examination survey (NHANES II) results. RESULTS: The odds ratio of the SRS male aged 45-75 compared to NHANES was 2.4 for pleura abnormalities and 0.8 for parenchymal abnormalities. Using logistic regression, the highest-risk worker division was construction (OR = 2.76); asbestos exposure was clearly associated with pleural abnormality (OR = 2.15). CONCLUSIONS: Pulmonary abnormalities were higher in former SRS workers than that in general population. Asbestos and possibly other exposures were related to pulmonary disease in this population. PMID- 16254951 TI - Methods for using narrative text from injury reports to identify factors contributing to construction injury. AB - BACKGROUND: Several methods exist for classifying injuries from written text, thereby identifying possible points of intervention. We describe an innovative method for such classification. METHODS: Using Haddon's matrix as a framework, two independent reviewers coded text from over 4,000 injury reports into a qualitative software package to identify factors contributing to injuries sustained during construction of Denver International Airport (DIA). We compared our classification scheme with three others. RESULTS: This process created a coded data set, an expanded version of Haddon's matrix adapted for construction injury, and coding rules for interpreting narrative text. Haddon's matrix provides a flexible theoretical framework for coding information about a spectrum of contributing factors. CONCLUSIONS: Narrative descriptions from injury reports can provide detail on circumstances surrounding injuries and identify factors contributing to injury. Forms guiding investigators to explicitly consider human, organizational, and environmental factors could foster more complete descriptions of factors contributing to construction injury. PMID- 16254952 TI - Facial nerve palsy after acute exposure to dichloromethane. AB - BACKGROUND: Dichloromethane poisoning affects predominantly the central nervous and the cardiovascular systems, and results from both carboxyhemoglobin formation and direct solvent-related narcosis. Exposure is frequently occupational and related to paint-stripping. Several reports have described severe adverse effects as well as fatalities. Conflicting reports regarding peripheral nerve toxicity have been found with no reports of clinical acute toxicity heretofore. METHODS: We present a case report of a patient who developed a facial nerve palsy after acute occupational exposure to Dichloromethane. The patient was part of a paint removing crew who have worked without proper protecting measures and were thus exposed to high levels of Dichoromethane. RESULTS: The patient was involved in paint stripping with Dichloromethane, and developed facial nerve palsy. Other known causes of facial palsy were excluded, and although idiopathic palsy cannot be ruled out, there were no corroborating findings. Carboxyhemoglobin levels taken after a significant delay were normal. CONCLUSION: This is the first article that describes a case of Facial Nerve Palsy related to acute dichloromethane exposure, indicating a possible peripheral neurotoxic effect of this solvent. PMID- 16254953 TI - Pade methods for reconstruction and feature extraction in magnetic resonance imaging. AB - Methods utilizing Pade approximants are investigated for implementation with magnetic resonance imaging data and are presented both for direct image reconstruction and for feature extraction. Pade approximants are a numerical tool that can be used to accelerate the convergence of a slowly converging sequence by estimating the fully converged sequence values from early data points. Pade approximants can be calculated directly from k-space data by solving a set of linear matrix equations to produce signal values for any desired location in the image domain. This gives an estimate of the fully converged signal intensity at each pixel location in the image, raising the possibility of reconstructing a better estimate of the object from a reduced data set. These methods have been tested on phantom and human data both for image reconstruction and for feature extraction. In image reconstruction, considerable convergence acceleration can be achieved, with steep intensity boundaries reproduced in keeping with higher resolution reconstructions and oscillatory truncation artifact characteristic of Fourier reconstruction removed. The convergence acceleration is variable and there is the possibility of fine detail suppression when insufficient data are included. The use of Pade methods as a tool for feature extraction has shown good agreement with extraction from high-resolution reference data. In this approach the edge information comes intrinsically from Pade reconstruction. PMID- 16254955 TI - Novel approach to the measurement of absolute cerebral blood volume using vascular-space-occupancy magnetic resonance imaging. AB - Quantitative determination of cerebral blood volume (CBV) is important for understanding brain physiology and pathophysiology. In this work, a novel approach is presented for accurate measurement of absolute CBV (aCBV) using vascular-space-occupancy (VASO) MRI, a blood-nulling pulse sequence, in combination with the T(1) shortening property of Gd-DTPA. Two VASO images with identical imaging parameters are acquired before and after contrast agent injection, resulting in a subtracted image that reflects the amount of blood present in the brain, i.e., CBV. With an additional normalizing factor, aCBV in units of milliliters of blood per 100 mL of brain can be estimated. Experimental results at 1.5 and 3 T systems showed that aCBV maps with high spatial resolution can be obtained with high reproducibility. The averaged aCBV values in gray and white matter were 5.5 +/- 0.2 and 1.4 +/- 0.1 mL of blood/100 mL of brain, respectively. Compared to dynamic susceptibility contrast techniques, VASO MRI is based upon a relatively straightforward theory and the calculation of CBV does not require measurement of an arterial input function. In comparison with previous pre/postcontrast difference approaches, VASO MRI provides maximal signal difference between pre- and postcontrast situation and does not require the use of whole blood for signal normalization. PMID- 16254954 TI - Real-time blood flow imaging using autocalibrated spiral sensitivity encoding. AB - A novel spiral phase contrast (PC) technique was developed for high temporal resolution imaging of blood flow without cardiac gating. An autocalibrated spiral sensitivity encoding (SENSE) method is introduced and used to reconstruct PC images. Numerical simulations and a flow phantom study were performed to validate the technique. To study the accuracy of the flow measurement in vivo, a high resolution cardiac experiment was performed and a subset of undersampled SENSE reconstructed data were reconstructed. Good agreement between the velocity measurement from the fully-sampled and undersampled data was achieved. Real-time experiments were performed to measure blood velocity in the ascending aorta and aortic valve, and during a Valsalva maneuver. The results demonstrate the potential of this technique for real-time flow imaging. PMID- 16254956 TI - Parallel magnetic resonance imaging using the GRAPPA operator formalism. AB - In this article it is shown that GRAPPA reconstruction can be reformulated as a matrix operator, similar to ladder or propagator operators used in quantum mechanics, that shifts data in k-space. Using this formalism, it is shown that there exists an infinitesimal GRAPPA operator that shifts data in k-space by arbitrarily small amounts. Other desired k-space shifts can then be accomplished through repeated applications of this infinitesimal GRAPPA operator. Implications of these ideas are described. PMID- 16254957 TI - Local delivery of bisphosphonate from coated orthopedic implants increases implants mechanical stability in osteoporotic rats. AB - Patients with osteoporosis and joint disabilities represent a constant growing and challenging population to be treated in the musculoskeletal clinical field. Especially in the case of total hip arthroplasty, new solutions should be developed to compensate for the double negative factors, peri-implant osteolysis, and osteoporotic bone loss, affecting the quality of implant outcome. The goal of this study was then to establish a proof of concept for orthopedic implant used as Zoledronate delivery in osteoporotic rats, and in particular, to verify if this approach could increase the initial implant stability. Twenty-five female 6 month-old Wistar rats were ovariectomized 6 weeks before the implantation to induce osteoporosis. The animals were randomly separated in five groups representing the different Zoledronate concentrations in the HA coating: 0, 0.2, 2.1, 8.5, and 16 microg/implant. Histomorphometric measures and peri-implant bone volume fraction were assessed and mechanical stability tests were performed. Bone volume fraction and biomechanical results clearly illustrate the positive effect of Zoledronate coated implants in the osteoporotic rats. A remarkable result was to show the existence of a window of Zoledronate content (0.2 to 8.5 microg/implant) in which the mechanical fixation of the implant increased. We were able to establish the proof of concept for orthopedic implants used as a drug delivery system in osteoporotic rats. The local bisphosphonate delivery from a calcium phosphate coating allowed increase of the mechanical fixation of an orthopedic implant. This study shows that orthopedic implants containing bisphosphonates could be beneficial for osteoporotic patients in need of a total joint replacement. PMID- 16254958 TI - Antifungal activity of DNA-lipid complexes and DNA-lipid films against Candida species. AB - In this study amphiphilic lipids, DNA-lipid complexes, and DNA-lipid films were prepared, and their antifungal activity against Candida species was examined. The amphiphilic lipids were synthesized from a reaction of glycine or L-alanine with n-alkyl alcohol in the presence of p-toluene sulfonic acid. DNA-lipid complexes, which were prepared by the simple mixing of DNA and amphiphilic lipids, were insoluble in water. Self-standing, water-insoluble DNA-lipid films were prepared by casting the DNA-lipid complexes from a chloroform/ethanol solution. The antifungal activities of the lipids and DNA-lipid complexes against the Candida species were evaluated by minimum inhibitory concentrations (MICs); those of DNA lipid films were evaluated by the disk diffusion method. The seven kinds of lipids, DNA-lipid complexes, and DNA-lipid films showed antifungal activity, and no differences were seen in the antifungal activities between glycine and L alanine derivatives. The lipids, DNA-lipid complexes, and DNA-lipid films, which have shorter alkyl chain length in lipids, showed antifungal activity against all Candida species. However, the effect of antifungal activity against Candida species decreased with increased alkyl chain length in lipids. In this study, it was found that lipids, DNA-lipid complexes, and films with a decyl or dodecyl group exhibit more favorable antifungal activity. PMID- 16254959 TI - Detection and characterization of human caliciviruses in hospitalized children with acute gastroenteritis in Blantyre, Malawi. AB - The human caliciviruses (HuCVs), including Norovirus and Sapovirus, are recognized causes of acute gastroenteritis in children and adults. A 1-year study was undertaken in Blantyre, Malawi, to examine the prevalence, and genetic diversity, of human caliciviruses (HuCVs) amongst children under 5 years of age hospitalized with acute gastroenteritis. Using the reverse transcription polymerase chain reaction (RT-PCR), combined with nucleotide sequencing of the RT PCR products, HuCVs were detected in 34/398 (8.5%) of children. Twelve (35.3%) of the children were co-infected with additional enteric viruses (predominantly rotavirus). The HuCVs comprised 26 Noroviruses (6.5%) and 8 Sapoviruses (2.0%). Each of the Noroviruses belonged to genogroup II, and could be further classified into six genotypes, including GII/3 (18 strains), GII/4 (2 strains), GII/11 (1 strain), GII/13 (1 strain), GII/16 (2 strains), and a putative new genotype GII/20 (2 strains). Each of the Sapoviruses belonged to genogroup GIII. HuCVs are the second most commonly identified viral enteropathogens (after rotavirus) among hospitalized children with gastroenteritis in Malawi. PMID- 16254960 TI - Measles virus genotype B2 is not inactive: evidence of continued circulation in Africa. AB - This study describes two measles outbreaks--one in Cape Town, South Africa in November 2002, and the other in Luanda, Angola in March 2003. The outbreaks were notable because they were caused by closely related genotype B2 viruses. This genotype was first described in an outbreak in Libreville, Gabon in the 1980s and was labeled as inactive by the World Health Organization in 2003 because it had not been detected for over 15 years. As the first three cases in the Cape Town outbreak were Angolan citizens who recently arrived from Angola, it appears likely that the source of the virus was Angola. Molecular analysis of specimens collected during the outbreak in Luanda provided direct evidence for the circulation of genotype B2 measles virus (MV) in Angola. This study clearly demonstrates that there is still active circulation of genotype B2 in Africa, and we propose that its apparent inactivity is merely the result of insufficient virologic/molecular surveillance in the region. These findings highlight the need for expanded molecular surveillance in Africa. PMID- 16254961 TI - Molecular characterization of a Canadian human parechovirus (HPeV)-3 isolate and its relationship to other HPeVs. AB - A new member of the Parechovirus genus (HPeV-3) has been recently isolated from a stool specimen of a young Japanese child with transient paralysis (isolate A308/99). Three additional HPeV-3 Canadian cases associated with severe neonatal infections were subsequently described by our group. At this time, limited information is available on the molecular characterization of the HPeV-3 genome. In this study, we report the complete genome sequence of a Canadian HPeV-3 isolate (Can82853-01) recovered in 2001 from the nasopharyngeal aspirate of a neonate with a sepsis-like syndrome. The Can82853-01 genome was 7,322 nucleotides (nt) in length and had 96.4%, 77.7%, and 77.5% nt identity with the A308/99 (HPeV 3), HPeV-1, and HPeV-2 reference strains, respectively. Furthermore, the Can82853 01 polyprotein had 98.2%, 86.9%, and 84.7% amino acids identity compared to the same viruses. Most differences between the polyproteins of HPeV-3 isolates and those of HPeV-1 and -2 were located in the VP0-VP3-VP1 capsid region whereas the non-structural proteins were relatively conserved. The primary and predicted secondary structures of 5'UTR and 3'UTR of the HPeV-3 isolates were also different than those of HPeV-1 and -2 reference strains. Phylogenetic analysis confirmed the relatedness between Can82853-01 and A308/99 and their divergence from other picornaviruses. PMID- 16254962 TI - Molecular characterization and genetic variation of the VP7 gene of human rotaviruses isolated in Paraguay. AB - Rotavirus is the main cause of acute diarrhea in infants and young children worldwide. In Paraguay, acute diarrhea ranks fourth among the causes of mortality in children under 4 years of age. Rotavirus was detected in 93 out of 410 patients admitted to three main hospitals in Asuncion, Paraguay from August 1998 to August 2000. Samples from 64 patients were analyzed by RT-PCR for G and P typing. G4P[8] (46.9%; 30/64) was the most common strain detected, followed by G9P[8] (17.2%; 11/64) and G1P[8] (10,9%; 7/64). Since G4 was predominant during the epidemiological peaks of 1998 and 1999, four samples were sequenced and all grouped into sublineage Ic. This sublineage was reported for the first time in 1998 in Argentina, southern border of Paraguay, and it was shown to be responsible for the increased prevalence of G4 during the epidemiological season of 1998 in that country. In addition, Paraguayan G1 strains grouped in different lineages (I and II). However, G9 was predominant during the rotavirus epidemiological peak of 2000, and phylogenetic analysis of five samples grouped into a common emergent/reemergent lineage that circulates worldwide. Since vaccination could reduce the severity and the number of cases of rotavirus disease, this study suggests that a vaccine containing recently isolated variants of the most prevalent types (G1-G4) together with the emerging G9 type, might be sufficient to elicit a protective immune response against the rotavirus types circulating currently in Paraguay. PMID- 16254963 TI - Identification of sapovirus infection among Japanese infants in a day care center. AB - A total of 921 fecal specimens collected from 44 infants in a day care center in Tokyo, Japan during June 1999 to July 2000 were tested for the presence of sapovirus by reverse transcription-polymerase chain reaction (RT-PCR). Of 88 fecal specimens from infants with acute gastroenteritis, 2.3% (2) were found to be positive for sapovirus. Twenty-two of 833 (2.6%) fecal specimens collected from asymptomatic infants were also infected with this virus. Another interesting feature was the demonstration of high incidence of sapovirus infection (95.5%, 21 of 22) identified in a single day care center, which was not due to viral shedding after the latest acute gastroenteritis. Sapovirus was subjected to genetic analysis by sequencing. Up to 4 of 24 sapoviruses (16.7%) were clustered into genogroup IV known as a rare group. Remarkably, majority (79.2%, 19 of 24) of sapovirus isolates detected in this study turned out to present one novel sapovirus genotype tentatively called GI/8. It was noteworthy to point out that the GI/8 sapovirus infection was apparently confined only within the period of 9 weeks (44th through 52nd weeks). This pattern of infection implied the outbreak of asymptomatic GI/8 sapovirus infection in these subjects. The findings clearly indicate genogroup I sapovirus can be classified into eight genotypes. This is the first report to underscore that sapovirus pathogen causes not only clinical manifestations of acute gastroenteritis but also asymptomatic infection in infants in day care centers in Japan. PMID- 16254964 TI - Predictive value of drug levels, HIV genotyping, and the genotypic inhibitory quotient (GIQ) on response to saquinavir/ritonavir in antiretroviral-experienced HIV-infected patients. AB - Plasma levels of HIV protease inhibitors (PI) are often close to IC50 values of wild-type viruses when administered without ritonavir boosting. The impact of drug levels, resistance mutations, and the genotypic inhibitory quotient (GIQ) were examined on the response to saquinavir/ritonavir (SQV/r)-based salvage therapy. Patients with prior exposure to PI other than SQV and currently failing virologically were recruited prospectively in a multicenter trial. All patients began SQV/r 1000/100 mg bid plus another two antiretrovirals. A total of 139 patients was enrolled. At month 12, virological response (VR), defined as plasma HIV-RNA decline >1 log, was recorded in 68.2% of patients on treatment (41.7% in the intent-to-treat analysis). The median baseline number of protease resistance mutations was three. The VR was significantly lower in patients with >5 protease resistance mutations and in those with plasma SQV Cmin<0.1 microg/ml. At week 48, the VR was seen in 77.1% of patients with a GIQ>0.04 compared to 18.2% of those with a lower GIQ (P=0.001). In the multivariate analysis, 0.1 microg/ml were independently associated with VR. Interestingly, drug levels had their highest predictive value of the VR at week 12, since low SQV plasma levels often permitted ruling out poorly adherent patients. In contrast, the number of protease resistance mutations had the highest impact on the VR at week 24, suggesting that for those taking the drugs, the VR is highly dependent of the presence of resistance mutations. At any time, nevertheless, the GIQ provided the most accurate prediction of the VR. PMID- 16254965 TI - Antigenicity and immunogenicity of the immunodominant region of hepatitis B surface antigen displayed on bacteriophage T7. AB - The immunodominant region of hepatitis B virus (HBV) located in the viral small surface antigen (S-HBsAg) elicits virus-neutralizing and protective antibodies. In order to develop an easy and inexpensive method to produce this region without the need for extensive purification, amino acid residues 111-156 of S-HBsAg were fused to the C-terminal end of the 10B capsid protein of T7 phage. Western blotting and ELISA confirmed the expression of the recombinant protein on the surface of the phage particles. The recombinant phage exhibited the antigenic and immunogenic characteristics of HBsAg, illustrating its potential as an immunological reagent and vaccine. PMID- 16254967 TI - Molecular characterization of the HA gene of influenza type B viruses. AB - Nucleotide sequences of the HA1 subunit of influenza B viruses isolated in Portugal between 1994 and 2003 influenza winter seasons were analyzed by the Neighbor-Joining algorithm and rates of HA1 evolution estimated by linear regression. From 1994 to 2002, all influenza B viruses studied were of the Yamagata lineage. Strains isolated from 1994 to 1996, 1996 to 1999, and 1999 to 2002 revealed a high similarity with B/Beijing/184/93, B/Yamanashi/166/98, and B/Sichuan/379/99, respectively, and strains isolated during 1994-1995, 1996-1997, and 1998-1999 clustered in more than one branch of the phylogenetic tree. Victoria-related strains reappeared during 2002/2003 and formed only one branch in the phylogenetic tree revealing a closer relationship to B/Shandong/7/97. Evolutionary rates for strains from the Yamagata lineage were estimated as 3.82x10(-3) nucleotides/site/year and 2.62x10(-3) nucleotides/site/year for Victoria-related strains. In order to identify putative influenza B HA1 codons under selective pressure, a codon-substitution model for heterogeneous selective pressure at amino acid sites was used. A percentage of 97.3% of codons under negative selective pressure and 2.7% of codons under positive selective pressure (omega=dN/dS=2.65) were estimated, with posterior probability higher than 0.90. Amino acid sites 75, 197, and 199 were found more likely to be under positive selective pressure. PMID- 16254966 TI - HIV infection and protease genetic diversity in a rural area of the Southern Central African Republic. AB - Thirty-eight of 175 (21.7%) consecutive women of childbearing age from a rural area near Bangassou were tested HIV-positive. Ten protease and two protease/reverse transcriptase sequences (31.5% samples) were obtained. Eight sequences clustered into subtype J, A, G; two sequences were 13_cpx recombinant forms and two were indeterminate. The high proportion of HIV found suggests a recent outbreak of diversified HIV strains. PMID- 16254968 TI - Anesthetist to patient transmission of hepatitis C virus associated with non exposure-prone procedures. PMID- 16254969 TI - Genetic heterogeneity of hepatitis E virus in Darfur, Sudan, and neighboring Chad. AB - The within-outbreak diversity of hepatitis E virus (HEV) was studied during the outbreak of hepatitis E that occurred in Sudan in 2004. Specimens were collected from internally displaced persons living in a Sudanese refugee camp and two camps implanted in Chad. A comparison of the sequences in the ORF2 region of 23 Sudanese isolates and five HEV samples from the two Chadian camps displayed a high similarity (>99.7%) to strains belonging to Genotype 1. But four isolates collected in one of the Chadian camps were close to Genotype 2. Circulation of divergent strains argues for possible multiple sources of infection. PMID- 16254970 TI - Emergence of G3 and G9 rotavirus and increased incidence of mixed infections in the southern region of Ireland 2001-2004. AB - Two hundred and thirty fecal specimens were collected from children (up to 5 years of age) admitted with suspected rotaviral gastroenteritis at four Irish hospitals (Cork University Hospital, Mercy Hospital, Cork, Waterford Regional Hospital, and Kerry General Hospital) in the southern region of Ireland, between 2001 and 2004. Following laboratory confirmation of the aetiological agent, the rotavirus G-type was determined in all positive samples by reverse transcriptase polymerase chain reaction (RT-PCR). The distribution of the G-types (n=230) over the 3 year period was G1 (31%), G9 (21.8%), G3 (8.7%), G4 (6.5%), and G2 (3.5%). There were many mixed infections which accounted for 28.5% of the collection. G9 emerged as the most prevalent G type (30.1%) in 2001-2002, whilst G3 first emerged in 2002-2003 and accounted for 15.8% of the collection. Notably, G2 strains were present at a very low frequency (3.5%) during 2001-2004, compared to an earlier study (1997-1999), where they accounted for 28.5% of the specimens. A smaller subset of the study collection was similarly P-typed (n=139). P[8]-type was identified as the most prevalent P-type, accounting for 97.4% (n=186), while P[4] accounted for just 2.6% (n=5) of the collection. The low frequency of P[4] coincided with the decrease in G2 strains in circulation. The key finding in this study was the emergence of G3- and G9-serotypes as epidemiologically important rotavirus strains since 1999, and the low prevalence of the previously common G2 strains in Ireland. The profile of rotavirus is changing continuously in Ireland and the implications for a successful vaccination program are discussed. PMID- 16254971 TI - Real-time NASBA detection of SARS-associated coronavirus and comparison with real time reverse transcription-PCR. AB - Severe acute respiratory syndrome (SARS) exhibits a high mortality rate and the potential for rapid epidemic spread. Additionally, it has a poorly defined clinical presentation, and no known treatment or prevention methods. Collectively, these factors underscore the need for early diagnosis. Molecular tests have been developed to detect SARS coronavirus (SARS-CoV) RNA using real time reverse transcription polymerase chain reaction (RT-PCR) with varying levels of sensitivity. However, RNA amplification methods have been demonstrated to be more sensitive for the detection of some RNA viruses. We therefore developed a real-time nucleic acid sequence-based amplification (NASBA) test for SARS-CoV. A number of primer/beacon sets were designed to target different regions of the SARS-CoV genome, and were tested for sensitivity and specificity. The performance of the assays was compared with RT-PCR assays. A multi-target real-time NASBA application was developed for detection of SARS-CoV polymerase (Pol) and nucleocapsid (N) genes. The N targets were found to be consistently more sensitive than the Pol targets, and the real-time NASBA assay demonstrates equivalent sensitivity when compared to testing by real-time RT-PCR. A multi target real-time NASBA assay has been successfully developed for the sensitive detection of SARS-CoV. PMID- 16254972 TI - Plasma total homocysteine and memory in the elderly: the Hordaland Homocysteine Study. AB - We examined the relation between plasma total homocysteine (tHcy), folate, vitamin B12, and episodic memory in elderly community-dwelling subjects. A population-based study was conducted in 1992 and 1993, and subjects were re investigated after 6 years. Plasma analytes were determined on both occasions. At follow-up, memory performance, using the Kendrick Object Learning Test, was investigated in 2,189 subjects (age, 65-67 years at baseline). Subjects with memory deficit (test score, < 25) had higher tHcy and lower folate at follow-up compared with those without memory deficit: 12.6 (95% confidence interval [CI], 12.1, 13.1) versus 11.5 (95% CI, 11.3, 11.6) micromol/L (p < 0.001) for tHcy, and 6.7 (95% CI, 6.2, 7.1) versus 7.6 (95% CI, 7.5, 7.8) nmol/L (p < 0.001) for folate. The risk of memory deficit increased according to quintiles of tHcy both at baseline and at follow-up. A decline in tHcy, or an increase in folate, over a 6-year period was associated with a higher memory test score; and vice versa. These findings indicate that increased plasma tHcy is an independent risk factor for memory deficit both cross-sectionally and prospectively, and that a "favorable" change in folate or tHcy concentrations over time is associated with better memory performance. PMID- 16254973 TI - Common variants of LRRK2 are not associated with sporadic Parkinson's disease. AB - Multiple mutations in the gene for the leucine-rich repeat kinase (LRRK2) cause autosomal dominant late-onset parkinsonism (PARK8). The Gly2019Ser mutation appears to be common in different populations. To investigate whether this novel gene influences the non-Mendelian sporadic form of Parkinson's disease, we genotyped 121 single nucleotide polymorphisms comprehensively covering the entire LRRK2 gene region in a set of 340 Parkinson's disease patients and 680 matched control subjects from Germany. No association could be demonstrated. We have therefore no evidence for the existence of a common variant in LRRK2 that has a strong influence on Parkinson's disease risk. PMID- 16254974 TI - Selective sequestration of X4 isolates by human genital epithelial cells: Implication for virus tropism selection process during sexual transmission of HIV. AB - X4 and R5 HIV strains are present in the semen of men infected with HIV but R5 isolates are transmitted preferentially. The role of human epithelial cells in this selection is addressed. Three human cervical cell lines-CaSki, SiHa, and HEC1A-and normal human vaginal cells from HIV-negative donors were characterized for HIV receptor expression and incubated with X4 and R5 laboratory-adapted strains or primary isolates. The infection was assessed by detection of intracellular HIV DNA. The three cell lines were shown to express on their surface the CXCR4 and GalCer molecules, but not the CD4 and CCR5 ones. The three cell lines and normal human vaginal cells were found to be selectively permissive to X4 HIV entry; the preincubation of the cell lines with rhSDF-1 inhibited this infection. The detection of the intracellular proviral DNA in the cell lines and in normal human vaginal cells demonstrated a selective integration of X4 strains. Additional experiments showed that no extracellular RNA was detected in the supernatants of HEC1A cells infected by X4 isolates either after 18 days of culture or after incubation with PHA-stimulated PBMCs and that no transmission occurred after co-culture between infected HEC1A cells and PHA-stimulated PBMCs. These results suggest specific sequestration of X4 strains by genital epithelial cells, which could explain, at least in part, the HIV tropism selection process during sexual intercourse. PMID- 16254975 TI - Comparison of the COBAS TaqMan HBV test with the COBAS Amplicor monitor test for measurement of hepatitis B virus DNA in serum. AB - Quantitation of low hepatitis B virus (HBV) DNA levels in patients with chronic hepatitis B is important for monitoring natural history of disease and treatment efficacy. This study aimed to compare the quantitation range and analytical sensitivity of the newly developed COBAS TaqMan HBV test (TaqMan test) with the COBAS Amplicor HBV Monitor Test (Amplicor test), using the Eurohep HBV reference plasma and serum samples from patients. Serial dilutions (2.7x10(1)-2.7x10(8) copies/ml) of the Eurohep HBV reference plasma and 50 serum samples from chronic hepatitis B patients were tested by both assays. The TaqMan test could detect seven (2.7x10(2)-2.7x10(8) copies/ml) of eight dilutions of the reference plasma, while the Amplicor test could only detect three of them (2.7x10(3)-2.7x10(5) copies/ml). The HBV DNA values measured by the TaqMan test correlated very well with the theoretical Eurohep standard values (r=0.998, P<0.001). There were good correlations between the HBV DNA levels measured by the two assays on both the Eurohep reference plasma (r=0.993, P<0.001) and serum samples from patients (r=0.904, P<0.001). Compared to the Amplicor test, the TaqMan test had a higher sensitivity (50 vs. 300 copies/ml), shorter assay time (6 vs. 10 hr), and wider dynamic range (8 vs. 3 logs), and was more cost-effective in a clinical setting. These data indicate that the TaqMan test is an excellent tool for HBV DNA quantitation. PMID- 16254976 TI - Evaluation of a total core antigen assay for the diagnosis of hepatitis C virus infection in hemodialysis patients. AB - Hemodialysis patients are recognized as a group at high risk of infection with hepatitis C virus (HCV). Therefore, such a population should be screened routinely for the presence of HCV viremia. Since nucleic acid techniques remain expensive and largely unavailable in many laboratories in the developing world, the present study assesses the clinical usefulness of the HCV core antigen enzyme immunoassay for the diagnosis of HCV infection in dialysis patients. One hundred seventy-five dialysis patients were screened for the presence of anti-HCV antibodies and HCV RNA in the serum. One hundred twenty-eight serum samples were collected from the 76 patients who were anti-HCV antibody- and/or HCV RNA positive. These were evaluated for total HCV core antigen. Of these samples, 55 had sufficient volume to be further tested to quantify HCV RNA by reverse transcription polymerase chain reaction (RT-PCR). Genotyping of the HCV strains showed that the majority belonged to genotype 1b (77%). The HCV core antigen assay showed a sensitivity and specificity of 84% and 89%, respectively. The use of core antigen assay has enabled the early detection of three patients who developed an acute hepatitis C infection during the period of study. A correlation study was undertaken between the quantitative values of viral load, expressed as pg/ml of HCV core antigen in serum, and viral RNA in UI/ml. A significant correlation was observed (Pearson's correlation coefficient: 0.552; P<0.001). In conclusion, detection of HCV core antigen in serum is an inexpensive, reliable, and highly specific assay that can be useful in most laboratory settings to diagnose HCV infection, and especially in laboratories where nucleic acid technologies are not yet available. PMID- 16254977 TI - DNA microarray technique for detection and identification of seven flaviviruses pathogenic for man. AB - A flavivirus microarray was developed for detection and identification of yellow fever (YF), West Nile, Japanese encephalitis (JE), and the dengue 1-4 viruses, which are causing severe human disease all over the world. The microarray was based on 500-nucleotide probe fragments from five different parts of the seven viral genomes. A low-stringent amplification method targeting the corresponding regions of the viral genomic RNA was developed and combined with hybridization to the microarray for detection and identification. For distinction of the generated virus-specific fluorescence-patterns a fitting analysis procedure was adapted. The method was verified as functional for all seven flaviviruses and the strategy for the amplification, combined with the long probes, provided a high tolerance for smaller genetic variability, most suitable for these rapidly changing RNA viruses. A potentially high detection and identification capacity was proven on diverged strains of West Nile and dengue viruses. The lower limit for detection was equivalent, or better, when compared to routinely used RT-PCR methods. The performance of the method was verified on human patient samples containing dengue viruses, or normal human serum spiked with YF or JE viruses. The results demonstrated the ability of the flavivirus microarray to screen simultaneously a sample for several viruses in parallel, in combination with a good lower limit of detection. PMID- 16254978 TI - Mutations on the FG surface loop of human papillomavirus type 16 major capsid protein affect recognition by both type-specific neutralizing antibodies and cross-reactive antibodies. AB - The aim of this study was to further characterize the conformational neutralizing epitopes present on the surface-exposed FG loop of human papillomavirus (HPV) type 16 L1 major capsid protein. We have generated previously two chimeric L1 proteins by insertion of a foreign peptide encoding an epitope of the hepatitis B core (HBc) antigen within the FG loop. In addition, three other chimeric L1 proteins were obtained by replacing three different FG loop sequences by the HBc motif and three others by point mutations. All these chimeric L1 proteins retained the ability to self-assemble into virus-like particles (VLPs), with the exception of the mutant with substitution of the L1 sequence 274-279 by the HBc motif. The eight chimeric VLPs were then analyzed for differential reactivity with a set of six HPV-16 and HPV-31 monoclonal antibodies that bound to conformational and linear epitopes. The binding patterns of these monoclonal antibodies confirmed that the FG loop contained or contributed to neutralizing conformational epitopes. The results obtained suggested that the H31.F7 antibody, an anti-HPV-31 cross-reacting and neutralizing antibody, recognized a conformational epitope situated before the 266-271 sequence. In addition, H16.E70 neutralizing antibody reactivity was reduced with L1 VLPs with an Asn to Ala point mutation at position 270, suggesting that Asn is a part of the epitope recognized by this antibody. This study contributes to the understanding of the antigenic structure of HPV-16 and -31 L1 proteins by confirming that the FG loop contributes to neutralizing epitopes and suggesting the existence of both type specific and cross-reactive conformational epitopes within the FG loop. PMID- 16254979 TI - G2 rotavirus infections in an infantile population of the South of Italy: variability of viral strains over time. AB - Rotavirus positive samples collected in Palermo, Italy, during 2002-2004 did not react with the G2 type-specific RV5:3 monoclonal antibodies (MAbs) and could be identified as G2 only by RT-PCR genotyping. The genetic variation of VP7 and VP4 antigenic proteins was studied in 14 G2 samples including a selection of both those successfully characterized by serotyping and those failing to be serotyped. The phylogenetic analysis performed on partial VP7 sequences showed a temporal clustering of these strains, with those isolated in Palermo in 2003 belonging to the same lineage of G2 MAbs-unreactive strains identified in UK in 1996-1997 and in Bari, Italy, in 2003-2004. A single amino acid substitution in VP7 antigenic region A, at position 96 (Asp-->Asn), was consistently associated with the loss of antigenic reactivity. Five of the G2 strains were further characterized by sequencing of VP4-encoding genes as belonging to the P[4] type, and separate lineages clustering the strains according to a temporal distribution could be described. VP7 and VP4 antigenic proteins analysis provided evidence that over the last 11 years, at least two different populations of G2P[4] rotavirus strains have been infecting the infant population in Palermo. Considering the role of anti-VP7 and anti-VP4 neutralizing antibodies in rotavirus immunity, the emergence of new VP7-VP4 gene combinations might influence rotavirus circulation in the infant population and should be taken into consideration when devising vaccination strategies. PMID- 16254980 TI - Changing prevalence of hepatitis B virus genotypes in Iceland. AB - At present eight hepatitis B virus (HBV) genotypes have been characterized: A to H. The most common genotype in Northern Europe is genotype A. So far there is no record of the specific HBV genotype distribution in Iceland. Iceland has a small population whose homogeneity has changed due to increasing migration during the past decades. The distribution of HBV genotypes in Iceland was analyzed using sera from 170 Icelandic patients. The samples were obtained before 1989, during an HBV epidemic among intravenous drug users in 1989 to 1992 and after 1994. A fragment of the HBV S-gene was amplified, sequenced and subjected to phylogenetic analysis. Among samples derived before 1989 genotypes A, C, and D were found. Most of the samples diagnosed during the epidemic belonged to genotype D and a smaller portion to genotype A. This suggests that the epidemic was most likely caused either by an endogenous HBV strain or by a strain imported from Europe or the USA. Among samples obtained after 1994, genotypes A to E and G were found, but the majority were of genotypes A, C, and D. This is consistent with an increase in migration and immigration from regions in Asia and Africa during the past 10 years. Thus, the changing prevalence of HBV genotypes in a small isolated community such as Iceland reflects the influence of migration and increasing contacts with regions outside the Western World. PMID- 16254983 TI - Changes in risk behavior and dynamics of hepatitis C virus infections among young drug users in Amsterdam, the Netherlands. AB - To elucidate the character and magnitude of the hepatitis C virus (HCV) epidemic among drug users in Amsterdam, 197 young drug users from the period 2000 to 2004 were compared with 215 counterparts from 1985 to 1989. Although injection risk behavior and HCV seroprevalence decreased sharply over time, HCV seroprevalence remains high (44%) among young drug users who have ever injected. Phylogenetic analysis shows that current HCV infections originate from diversification of strains already circulating in the past, but also from the recent introduction of new subtypes. HCV subtypes 1a and 3a remain the most prevalent among drug users in Amsterdam, but other subtypes such as 4d and 2b have entered the population. In conclusion, both the unpopularity of injecting drug use and the success of prevention campaigns are likely to be responsible for the decline in the seroprevalence of HCV and increased median time to seroconversion. Treatment of those infected chronically, in combination with the continuation of prevention programs, might decrease future HCV transmission. PMID- 16254981 TI - High prevalence of dual or triple infection of hepatitis B, C, and delta viruses among patients with chronic liver disease in Mongolia. AB - Mongolia is known for its high endemicity for hepatitis B virus (HBV), hepatitis C virus (HCV), and hepatitis delta virus (HDV) infections among apparently healthy individuals. However, there are little or no data on the prevalence and genotype distribution of HBV, HCV, and HDV among patients with chronic liver disease in Mongolia. Therefore, serum samples obtained in 2004 from 207 patients (age, mean+/-standard deviation, 51.0+/-11.9 years) including those with chronic hepatitis (n=90), liver cirrhosis (n=41), and hepatocellular carcinoma (n=76) were tested for serological and molecular markers of HBV, HCV, and HDV infections. Of the 207 patients, 144 (69.6%), 106 (51.2%), and 117 (56.5%) tested positive for hepatitis B surface antigen (HBsAg) and/or HBV DNA, HCV RNA, and HDV RNA, respectively. Collectively, 172 patients (83.1%) were viremic for one or more of these viruses, including dual viremia of HBV/HDV (26.6%) or HBV/HCV (7.7%) and triple HBV/HCV/HDV viremia (30.0%). Of note, triple ongoing infection was significantly more frequent among patients with hepatocellular carcinoma than among those with chronic hepatitis (63.2% vs. 14.4%, P<0.0001). One hundred sixty patients (77.3%) had a history of blood transfusion and/or surgery. The distribution of HBV genotypes among the 116 HBV-viremic patients was: A (0.9%), B (0.9%), C (6.0%), D (88.8%), and C plus D (3.4%). All 117 HDV isolates were classified into genotype I. The 106 HCV RNA-positive samples were typed as genotype 1b (92.5%), 2a (0.9%), or 1b plus 2a (6.6%); mixed infection of two distinct HCV genotypes was found exclusively in the patients with hepatocellular carcinoma. PMID- 16254985 TI - Evolutionary transformations of fetal membrane characters in Eutheria with special reference to Afrotheria. AB - Analysis of molecular data sets has provided new insights into higher-level relationships of living Eutheria, including the recognition of Afrotheria as a novel taxon. This offers an opportunity to take a fresh look at the evolution of organ systems, including some that are little used in traditional systematics. In the present study, we attempted a reconstruction of the evolution of characters associated with placentation, the fetal membranes and the female reproductive tract. The evolutionary history of 21 characters has been traced, based on a current hypothesis of eutherian relationships, by applying the computer program MacClade. Accordingly, the analysis provides a first comprehensive interpretation of the stem species pattern of Eutheria. Of particular note, this pattern includes an endotheliochorial chorioallantoic placenta. The reconstructed pattern of Eutheria does not change in the basal nodes of the group. Thus, no character transformations occur on the stem lineages of Laurasiatheria or Euarchontoglires, and even Afrotheria has mostly plesiomorphic character conditions. However, two character transformations occur on the common stem lineage of Afrotheria and its sister taxon Xenarthra, i.e., amniogenesis by cavitation instead of folding and the precocial state of the newborn. In addition, we recognized one character transformation on the stem lineage of Afrotheria, i.e., the occurrence of a four lobed allantoic sac. Thus, contrary to previous assertions, it is possible to identify morphological characters that could be synapomorphic for this novel taxon. PMID- 16254984 TI - A BAC library of the East African haplochromine cichlid fish Astatotilapia burtoni. AB - A BAC library was constructed from Astatotilapia burtoni, a haplochromine cichlid that is found in Lake Tanganyika, East Africa, and its surrounding rivers. The library was generated from genomic DNA of blood cells and comprises 96,768 individual clones. Its median insert size is 150 kb and the coverage is expected to represent about 14 genome equivalents. The coverage evaluation was based on genome size estimates that were obtained by flow cytometry. In addition, hybridization screens with five probes largely corroborate the above coverage estimate, although the number of clones ranged from 5 to 22 authenticated clones per single copy probe. The BAC library described here is expected to be useful to the scientific community interested in cichlid genomics as an important resource to gain new insights into the rapid evolution of the great species diversity of haplochromine cichlid fishes. PMID- 16254986 TI - Developmental constraints vs. variational properties: How pattern formation can help to understand evolution and development. AB - This article suggests that apparent disagreements between the concept of developmental constraints and neo-Darwinian views on morphological evolution can disappear by using a different conceptualization of the interplay between development and selection. A theoretical framework based on current evolutionary and developmental biology and the concepts of variational properties, developmental patterns and developmental mechanisms is presented. In contrast with existing paradigms, the approach in this article is specifically developed to compare developmental mechanisms by the morphological variation they produce and the way in which their functioning can change due to genetic variation. A developmental mechanism is a gene network, which is able to produce patterns in space though the regulation of some cell behaviour (like signalling, mitosis, apoptosis, adhesion, etc.). The variational properties of a developmental mechanism are all the pattern transformations produced under different initial and environmental conditions or IS-mutations. IS-mutations are DNA changes that affect how two genes in a network interact, while T-mutations are mutations that affect the topology of the network itself. This article explains how this new framework allows predictions not only about how pattern formation affects variation, and thus phenotypic evolution, but also about how development evolves by replacement between pattern formation mechanisms. This article presents testable inferences about the evolution of the structure of development and the phenotype under different selective pressures. That is what kind of pattern formation mechanisms, in which relative temporal order, and which kind of phenotypic changes, are expected to be found in development. PMID- 16254987 TI - Anatomy of the soul as reflected in the cerebral hemispheres: neural circuits underlying voluntary control of basic motivated behaviors. AB - Understanding the principles of cerebral hemisphere neural network organization is essential for understanding the biological foundations of cognition and affect thinking and feeling. A tripartite model of cerebral structure-function organization is reviewed, with attention focused on a behavior control system differentiation that mediates voluntary influences on three fundamental classes of goal-oriented behavior common to all animals. The model postulates just three cerebral divisions, one cortical and two nuclear (lateral or striatal, and medial or pallidal), that together generate a triple descending projection to the brainstem/cord motor system. This minimal circuit element is topographically organized and regionally differentiated, with the map of cortical areas serving as a basic starting point. Virtually all of the cerebral hemisphere projects on the upper brainstem behavior control column, atop the motor system hierarchy. The latter's rostral segment helps control ingestive (eating and drinking), defensive (fight or flight), and reproductive (sexual and parental) motivated behaviors, whereas its caudal segment helps control foraging or exploratory behavior to obtain or avoid specific goal objects associated with all classes of motivated behavior. PMID- 16254988 TI - Chemistry of the mind: neurochemical modulation of prefrontal cortical function. AB - The neurochemical modulation of prefrontal cortical function is reviewed with special reference to the ascending dopaminergic and serotoninergic projections. Evidence is surveyed from studies of rats, nonhuman primates, and humans to suggest that prefrontal dopamine has specific functions in attentional control and working memory, mediated mainly through the D1 receptor, whereas manipulations of serotonin are shown by contrast to affect reversal learning in monkeys and human volunteers and measures of impulsivity in rats. These findings are discussed in the context of these as well as other neurotransmitter systems (including noradrenaline and acetylcholine) having distinct roles in the neuromodulation of prefrontal cortical function. The capacity of the prefrontal cortex itself to exert top-down regulation of these ascending neurochemical systems is also discussed. PMID- 16254989 TI - Frontotemporal and dopaminergic control of idea generation and creative drive. AB - This article presents a three-factor anatomical model of human idea generation and creative drive, focusing on interactions between the temporal lobes, frontal lobes, and limbic system. Evidence is drawn from functional imaging, drug studies, and lesion analysis. Temporal lobe changes, as in hypergraphia, often increase idea generation, sometimes at the expense of quality. Frontal lobe deficits may decrease idea generation, in part because of rigid judgments about an idea's worth. These phenomena are clearest in verbal creativity, and roughly parallel the pressured communication of temporal lobe epilepsy, mania, and Wernicke's aphasia-compared to the sparse speech and cognitive inflexibility of depression, Broca's aphasia, and other frontal lobe lesions. The phenomena also shape non-linguistic creativity, as in that of frontotemporal dementia. The appropriate balance between frontal and temporal activity is mediated by mutually inhibitory corticocortical interactions. Mesolimbic dopamine influences novelty seeking and creative drive. Dopamine agonists and antagonists have opposite effects on goal-directed behavior and hallucinations. Creative drive is not identical to skill-the latter depends more on neocortical association areas. However, drive correlates better with successful creative output than skill does. Traditional neuroscientific models of creativity, such as the left brain - right brain hemispheric model, emphasize skills primarily, and stress art and musical skill at the expense of language and mathematics. The three-factor model proposed here predicts findings in a broad range of normal and pathological states and can be tested in many experimental paradigms. PMID- 16254991 TI - Identifying hypothalamic pathways controlling food intake, body weight, and glucose homeostasis. AB - The past decade has greatly increased our understanding and appreciation of the ability of the central nervous system (CNS) to regulate food intake and body weight. This was spearheaded by the discovery of key molecules regulating body weight homeostasis. It is now also apparent that the CNS, especially the hypothalamus, plays a primary role in directly regulating glucose homeostasis, independently of effects on body weight. These discoveries are important given the increasing incidences of obesity and type II diabetes in Western societies. In this article, we will highlight recent data from genetically modified mice. These data and other models have helped to dissect the CNS pathways regulating body weight and glucose homeostasis. Finally, although these studies have been illustrative, they also underscore our relative lack of knowledge and highlight the need for more definitive approaches to unravel the functional significance of these pathways. PMID- 16254992 TI - Generalized arousal of mammalian central nervous system. AB - A fundamental capacity of the mammalian CNS is becoming amenable to study with the techniques of functional genomics. Emphasized in this review are ascending connections from the medullary reticular formation and descending connections from the paraventricular nucleus of the hypothalamus. In particular, sex hormone effects on neurons allow us to relate generalized arousal to a specific form of arousal which is required for reproductive behaviors. PMID- 16254990 TI - Forebrain substrates of reward and motivation. AB - Electrical stimulation of the medial forebrain bundle can reward arbitrary acts or motivate biologically primitive, species-typical behaviors like feeding or copulation. The subsystems involved in these behaviors are only partially characterized, but they appear to transsynaptically activate the mesocorticolimbic dopamine system. Basal function of the dopamine system is essential for arousal and motor function; phasic activation of this system is rewarding and can potentiate the effectiveness of reward-predictors that guide learned behaviors. This system is phasically activated by most drugs of abuse and such activation contributes to the habit-forming actions of these drugs. PMID- 16254993 TI - Micturition and the soul. AB - There is a close connection between micturition and emotion. Several species use micturition to signal important messages as territorial demarcation and sexual attraction. For this reason, micturition is coordinated not in the spinal cord but in the brainstem, where it is closely connected with the limbic system. In cat, bladder afferents terminate in a cell group in the lateral dorsal horn and lateral part of the intermediate zone. Neurons in this cell group project to supraspinal levels, not to the thalamus but to the central periaqueductal gray (PAG). Neurons in the lateral PAG, not receiving direct sacral cord afferents, project to the pontine micturition center (PMC). The PMC projects directly to the parasympathetic bladder motoneurons and to sacral GABA-ergic and glycinergic premotor interneurons that inhibit motoneurons in Onuf's nucleus innervating the external striated bladder sphincter. Thus, PMC stimulation causes bladder contraction and bladder sphincter relaxation, i.e., complete micturition. Other than the PAG, only the preoptic area and a cell group in the caudal hypothalamus project directly to the PMC. The ventromedial upper medullary tegmentum also sends projections to the PMC, but they are diffuse and also involve structures that adjoin the PMC. Neuroimaging studies in humans suggest that the systems controlling micturition in cat and human are very similar. It seems that the many structures in the brain that are known to influence micturition use the PAG as relay to the PMC. This basic organization has to be kept in mind in the fight against overactive bladder (OAB) and urge-incontinence. PMID- 16254994 TI - Homeostatic, circadian, and emotional regulation of sleep. AB - A good night's sleep is one of life's most satisfying experiences, while sleeplessness is stressful and causes cognitive impairment. Yet the mechanisms that regulate the ability to sleep have only recently been subjected to detailed investigation. New studies show that the control of wake and sleep emerges from the interaction of cell groups that cause arousal with other nuclei that induce sleep such as the ventrolateral preoptic nucleus (VLPO). The VLPO inhibits the ascending arousal regions and is in turn inhibited by them, thus forming a mutually inhibitory system resembling what electrical engineers call a "flip-flop switch." This switch may help produce sharp transitions between discrete behavioral states, but it is not necessarily stable. The orexin neurons in the lateral hypothalamus may help stabilize this system by exciting arousal regions during wakefulness, preventing unwanted transitions between wakefulness and sleep. The importance of this stabilizing role is apparent in narcolepsy, in which an absence of the orexin neurons causes numerous, unintended transitions in and out of sleep and allows fragments of REM sleep to intrude into wakefulness. These influences on the sleep/wake system by homeostatic and circadian drives, as well as emotional inputs, are reviewed. Understanding the pathways that underlie the regulation of sleep and wakefulness may provide important insights into how the cognitive and emotional systems interact with basic homeostatic and circadian drives for sleep. PMID- 16254995 TI - Adaptive gain and the role of the locus coeruleus-norepinephrine system in optimal performance. AB - Historically, the locus coeruleus-norepinephrine (LC-NE) system has been implicated in arousal, but recent findings suggest that this system plays a more complex and specific role in the control of behavior than investigators previously thought. We review neurophysiological, anatomical, and modeling studies in monkey that support a new theory of LC-NE function. LC neurons exhibit two modes of activity, phasic and tonic. Phasic LC activation is driven by the outcome of task-related decision processes and is proposed to facilitate ensuing behaviors and to help optimize task performance. When utility in the task wanes, LC neurons exhibit a tonic activity mode, associated with disengagement from the current task and a search for alternative behaviors. Monkey LC receives prominent, direct inputs from the anterior cingulate (ACC) and orbitofrontal cortices (OFC), both of which are thought to monitor task-related utility. We propose that these prefrontal areas produce the above patterns of LC activity to optimize the utility of performance on both short and long time scales. PMID- 16254996 TI - Neural mechanisms of freezing and passive aversive behaviors. AB - Cues that predict aversive outcomes often produce marked inhibitions of behavior known as freezing, but it is unknown exactly what neural pathways cause this inhibition. The amygdala and bed nucleus of the stria terminalis, along with their projections to the periaqueductal gray, are strongly implicated in freezing, but it is not known how these structures inhibit motor output. The median raphe nucleus (MRN), which contains a major population of serotonin neurons, has also been implicated in freezing, but the serotonin neurons themselves do not seem to be involved, leaving it uncertain which neurons in this area promote freezing. Our recent work suggests that GABAergic neurons just lateral to the MRN, but not within the MRN, regulate freezing via projections to midbrain dopamine neurons. Because freezing pathways may control a variety of other passive aversive behaviors, their elucidation may help understand the mechanisms of addictions and compulsions, which involve a failure of aversive outcomes to inhibit behavior. PMID- 16254997 TI - Neural mechanisms of autonomic, affective, and cognitive integration. AB - Influential theoretical models propose a central role for afferent information from the body in the expression of emotional feeling states. Feedback representations of changing states of bodily arousal influence learning and facilitate concurrent and prospective decision-making. Functional neuroimaging studies have increased understanding of brain mechanisms that generate changes in autonomic arousal during behavior and those which respond to internal feedback signals to influence subjective feeling states. In particular, anterior cingulate cortex is implicated in generating autonomic changes, while insula and orbitofrontal cortices may be specialized in mapping visceral responses. Independently, ventromedial prefrontal cortex is recognized to support processes of internal (self-) reference that predominate in states of rest and disengagement and which putatively serve as a benchmark for dynamic interactions with the environment. Lesion data further highlight the integrated role of these cortical regions in autonomic and motivational control. In computational models of control, forward (efference copies) and inverse models are proposed to enable prediction and correction of action and, by extension, the interpretation of the behavior of others. It is hypothesized that the neural substrate for these processes during motivational and affective behavior lies within the interactions of anterior cingulate, insula, and orbitofrontal cortices. Generation of visceral autonomic correlates of control reinforce experiential engagement in simulatory models and underpin concepts such as somatic markers to bridge the dualistic divide. PMID- 16254998 TI - Intrinsic brain activity sets the stage for expression of motivated behavior. AB - Research in many species has provided increasingly detailed information on relevant, primarily subcortical brain systems supporting the expression of basic appetites and drives. While basic appetites and drives are essential for adaptation and survival in any environment, they are naturally constrained by an organism's inherent biology and modulated as circumstances dictate. The brain mechanisms which serve to constrain and modulate them, however, remain much less well understood. We suggest that the manner in which such constraint and potential modulation is achieved likely involves processes that emerge from the coordinated behavior of multiple brain systems, and functional brain imaging techniques such as PET and fMRI are beginning to help us understand aspects of such coordination. In this review we argue that, in pursuit of this understanding, we must focus not only on changes evoked in brain systems during various behaviors, but also on the ongoing and very costly intrinsic activity within these systems, for the latter may be at least as important as the evoked activity in terms of brain function in general and the constraint and modulation of basic appetites and drives in particular. Distinguishing intrinsic from evoked activity in the context of functional brain imaging experiments is challenging, however. Here we review some evolving strategies for doing so. PMID- 16254999 TI - The anatomy of the soul. Proceedings of a meeting, Ameland, The Netherlands, May 19-24, 2005. PMID- 16255000 TI - Relevance of an evidence-based ejaculation time cutoff point for neurobiological research of premature ejaculation. AB - The Diagnostic and Statistical Manual (DSM)-IV definition of premature ejaculation is not based on evidence-based studies. In particular, the absence of a well-defined quantitative measure of the intravaginal ejaculation latency time (IELT) makes the DSM definition inadequate. Therefore, the DSM-IV definition should be replaced by a medical definition that incorporates both quantitative as qualitative parameters of premature ejaculation. An evidence-based medical definition should include a cutoff point of the IELT at the 0.5 and 2.5 percentiles of the IELT distribution in the general male population. Such a definition has recently been proposed on the basis of a stopwatch study of the IELT in 491 men from five different countries. Similarly, a cutoff point of ejaculation frequency in laboratory rats enhances the probability to distinguish genuine rapid-ejaculator rats. Only by the strict application of these cutoff points is the probability enhanced that human and animal neurobiological research can prove whether these sexual endophenotypes differ in brain activation, have particular genetic genotypes (polymorphisms), are strictly under genetic control, or are dependent on environmental conditions and/or genotypic/environmental interactions. PMID- 16255001 TI - Romantic love: an fMRI study of a neural mechanism for mate choice. AB - Scientists have described myriad traits in mammalian and avian species that evolved to attract mates. But the brain mechanisms by which conspecifics become attracted to these traits is largely unknown. Yet mammals and birds express mate preferences and make mate choices, and data suggest that this "attraction system" is associated with the dopaminergic reward system. It has been proposed that intense romantic love, a cross-cultural universal, is a developed form of this attraction system. To determine the neural mechanisms associated with romantic love we used functional magnetic resonance imaging (fMRI) and studied 17 people who were intensely "in love" (Aron et al. [2005] J Neurophysiol 94:327-337). Activation specific to the beloved occurred in the right ventral tegmental area and right caudate nucleus, dopamine-rich areas associated with mammalian reward and motivation. These and other results suggest that dopaminergic reward pathways contribute to the "general arousal" component of romantic love; romantic love is primarily a motivation system, rather than an emotion; this drive is distinct from the sex drive; romantic love changes across time; and romantic love shares biobehavioral similarities with mammalian attraction. We propose that this attraction mechanism evolved to enable individuals to focus their mating energy on specific others, thereby conserving energy and facilitating mate choice-a primary aspect of reproduction. Last, the corticostriate system, with its potential for combining diverse cortical information with reward signals, is an excellent anatomical substrate for the complex factors contributing to romantic love and mate choice. PMID- 16255002 TI - A proposed hypothalamic-thalamic-striatal axis for the integration of energy balance, arousal, and food reward. AB - We elaborate herein a novel theory of basal ganglia function that accounts for why palatable, energy-dense foods retain high incentive value even when immediate physiological energy requirements have been met. Basal ganglia function has been studied from the perspective of topographical segregation of processing within parallel circuits, with primary focus on motor control and cognition. Recent findings suggest, however, that the striatum can act as an integrated unit to modulate motivational state. We describe evidence that the striatal enkephalin system, which regulates the hedonic impact of preferred foods, undergoes coordinated gene expression changes that track current motivational state with regard to food intake. Striatal enkephalin gene expression is also downregulated by an intrastriatal infusion of a cholinergic muscarinic antagonist, a manipulation that greatly suppresses food intake. To account for these findings, we propose that signaling through a hypothalamic-midline thalamic-striatal axis impinges on the cholinergic interneurons of the striatum, which via their large, overlapping axonal fields act as a network to modulate enkephalin-containing striatal output neurons. A key relay in this circuit is the paraventricular thalamic nucleus, which receives convergent input from orexin-coded hypothalamic energy-sensing and behavioral state-regulating neurons, as well as from circadian oscillators, and projects to cholinergic interneurons throughout the striatal complex. We hypothesize that this system evolved to coordinate feeding and arousal, and to prolong the feeding central motivational state beyond the fulfillment of acute energy needs, thereby promoting "overeating" and the consequent development of an energy reserve for potential future food shortages. PMID- 16255003 TI - Free will versus survival: brain systems that underlie intrinsic constraints on behavior. AB - This article discusses the neuroanatomical systems involved in the related functions of fear and discernment of the consequences of one's actions, two internal constraints on free will and action. Both mechanisms are related to a system for control and modulation of visceral function stretching from the spinal cord to the ventromedial prefrontal cortex, including the ventral striatum, ventral pallidum, and mediodorsal thalamus, the amygdala, the hypothalamus, the periaqueductal gray (PAG), and the brainstem reticular formation and autonomic nuclei. Reflexes at the lower levels provide rapid visceral and somatic reactions to threatening stimuli, while the PAG and hypothalamus coordinate these to produce more concerted responses. The amygdala interacts with the cortical sensory systems in the assessment of fear-related stimuli and modulates the reflex responses through projections to the hypothalamus and brainstem. The ventromedial prefrontal cortex, especially the "medial prefrontal network," is connected to the amygdala, hypothalamus, and PAG, and allows cortical control over the system in relation to a wider set of emotions. This cortical region is involved both in the assessment of reward and in mood disorders and it plays a central role in the ability to discern the consequences of one's actions and make appropriate behavioral choices. It also forms an interconnected circuit with specific cortical areas in the rostral superior temporal cortex, posterior parahippocampal cortex, and retrosplenial/posterior cingulate cortex. The overall function of this circuit is unclear, but may be involved in introspective monitoring of the individual. PMID- 16255004 TI - Ventromedial medulla: pain modulation and beyond. AB - The midbrain periaqueductal gray (PAG) and ventromedial medulla (VMM) are generally viewed as the core of an endogenous descending modulatory system. However, available data demonstrate that PAG and VMM do not specifically target nociceptive transmission and that activation of either structure affects numerous homeostatic physiological processes. Pseudorabies virus (PRV) is a useful tracer that is retrogradely and transynaptically transported. PRV injections into homeostatic effector organs invariably label VMM neurons, both serotonergic and nonserotonergic. Studies in anesthetized rats have implicated two types of nonserotonergic VMM neurons in nociceptive modulation: ON cells are thought to facilitate nociception and OFF cells to inhibit nociception. Yet, in the unanesthetized animal, the discharge of VMM neurons changes in response to innocuous stimuli and during situations unrelated to nociception. In particular, VMM cells appear to modulate the timing of micturition, with ON cells promoting the initiation of voiding and OFF cells promoting urine storage. VMM cells also modulate sensory transmission. During both micturition and sleep, OFF cells discharge and sensory responsiveness is depressed. In sum, the VMM is hypothesized to modulate spinal sensory, autonomic, and motor circuits in order to maintain homeostasis. PMID- 16255005 TI - Role of Barrington's nucleus in micturition. AB - Barrington's nucleus is a central component of the micturition circuit. This nucleus projects axons to the sacral parasympathetic nucleus, where preganglionic neurons innervating the urinary bladder are located. To clarify the functional role of this nucleus, the firing properties of Barrington's neurons that project axons to the spinal cord were examined. Based on these studies, a model begins to emerge that places Barrington's nucleus in the micturition pathway that is involved in increasing bladder pressure rapidly and strongly, while also maintaining high bladder pressure. In addition, Barrington's neurons are suggested to have another role, that is, increasing the probability of micturition contraction by activating a spinal excitatory pathway or disinhibiting a spinal inhibitory mechanism. In contrast to the excitatory role of Barrington's nucleus, this nucleus does not seem to trigger bladder relaxation. PMID- 16255006 TI - Functional imaging and the central control of the bladder. AB - The central control of the bladder is a complex, multilevel process. Recent advances in functional brain imaging have allowed research into this control in humans. This article reviews the functional imaging studies published to date and discusses the regions of the brain that have been implicated in the central control of continence. Brain regions that have been implicated include the pons (pontine micturition center, PMC), periaqueductal gray (PAG), thalamus, insula, anterior cingulate gyrus, and prefrontal cortices. The PMC and the PAG are thought to be key in the supraspinal control of continence and micturition. Higher centers such as the insula, anterior cingulate gyrus, and prefrontal regions are probably involved in the modulation of this control and cognition of bladder sensations, and in the case of the insula and anterior cingulate, modulation of autonomic function. Further work should aim to examine how the regions interact to achieve urinary continence. PMID- 16255007 TI - Human brain activation during sexual stimulation of the penis. AB - Penile sensory information is essential for reproduction, but almost nothing is known about how sexually salient inputs from the penis are processed in the brain. We used positron emission tomography to measure regional cerebral blood flow (rCBF) during various stages of male sexual performance. Compared to a passive resting condition (without penile erection), sexual stimulation of the penis increased rCBF in an area of the right hemisphere encompassing the posterior insula and adjacent posterior part of the secondary somatosensory cortex (SII) and decreased rCBF in the right amygdala. No activation was observed in either the thalamus, genital part of primary somatosensory cortex (SI), or hypothalamus. Based on these results we put forward the concept that during sexual performance the salience of the stimulus, represented by activation of the insula and SII, is of greater significance than the exact location of the stimulus, encoded in SI. The absence of activation in the hypothalamus indicates that this region is more important for the onset of sexual arousal than for the resulting sexual performance. Deactivation of the amygdala during sexual stimulation of the penis corresponds with a decrease of vigilance during sexual performance. PMID- 16255008 TI - Neural control of ejaculation. AB - Ejaculation is the most reinforcing component of sexual behavior. However, the neural substrates mediating ejaculation and processing ejaculation-related signals remain poorly understood. We review the current understanding of central control of ejaculation. Specifically, the recent identification of a candidate spinothalamic pathway involved in relay of ejaculation-specific signals is discussed. In addition, the discovery of a neural population of lumbar interneurons playing an pivotal role in expression of ejaculation is reviewed. PMID- 16255009 TI - Anatomy and neurochemistry of the pair bond. AB - Studies in monogamous rodents have begun to elucidate the neural circuitry underlying the formation and maintenance of selective pair bonds between mates. This research suggests that at least three distinct, yet interconnected, neural pathways interact in the establishment of the pair bond. These include circuits involved in conveying somatosensory information from the genitalia to the brain during sexual activity, the mesolimbic dopamine circuits of reward and reinforcement, and neuropeptidergic circuits involved specifically in the processing of socially salient cues. Here we present an integrated description of the interaction of these circuits in a model of pair bond formation in rodents with a discussion of the implications of these findings for evolution, individual variation, and human bonding. PMID- 16255010 TI - Low-dose prednisolone in addition to the initial disease-modifying antirheumatic drug in patients with early active rheumatoid arthritis reduces joint destruction and increases the remission rate: a two-year randomized trial. AB - OBJECTIVE: To assess the efficacy of low-dose prednisolone on joint damage and disease activity in patients with early rheumatoid arthritis (RA). METHODS: At the start of their initial treatment with a disease-modifying antirheumatic drug (DMARD), patients with early (duration < or =1 year) active RA were randomly assigned to receive either 7.5 mg/day prednisolone or no prednisolone for 2 years. Radiographs of the hands and feet were obtained at baseline and after 1 and 2 years and scored according to the Sharp score as modified by van der Heijde. Remission was defined as a Disease Activity Score in 28 joints of <2.6. Bone mineral density was measured by dual x-ray absorptiometry at baseline and after 2 years. RESULTS: Of the 250 patients included, 242 completed the study and 225 had radiographs available both at baseline and at 2 years. At 2 years, the median and interquartile range (IQR) change in total Sharp score was lower in the prednisolone group than in the no-prednisolone group (1.8 [IQR 0.5-6.0] versus 3.5 [IQR 0.5-10]; P = 0.019). In the prednisolone group, there were fewer newly eroded joints per patient after 2 years (median 0.5 [IQR 0-2] versus 1.25 [IQR 0 3.25]; P = 0.007). In the prednisolone group, 25.9% of patients had radiographic progression beyond the smallest detectable difference compared with 39.3% of patients in the no-prednisolone group (P = 0.033). At 2 years, 55.5% of patients in the prednisolone group had achieved disease remission, compared with 32.8% of patients in the no-prednisolone group (P = 0.0005). There were few adverse events that led to withdrawal. Bone loss during the 2-year study was similar in the 2 treatment groups. CONCLUSION: Prednisolone at 7.5 mg/day added to the initial DMARD retarded the progression of radiographic damage after 2 years in patients with early RA, provided a high remission rate, and was well tolerated. Therefore, the data support the use of low-dose prednisolone as an adjunct to DMARDs in early active RA. PMID- 16255011 TI - Very low-dose prednisolone in early rheumatoid arthritis retards radiographic progression over two years: a multicenter, double-blind, placebo-controlled trial. AB - OBJECTIVE: To assess the effect of 5 mg/day prednisolone on disease progression in patients with early rheumatoid arthritis (RA) receiving standardized disease modifying antirheumatic drug (DMARD) therapy. METHODS: Patients with active RA of <2 years' duration were randomly assigned in a double-blinded manner to receive prednisolone or placebo while starting concomitant DMARD therapy (gold sodium thiomalate or methotrexate). Hand and foot radiographs were taken at baseline and at 6, 12, and 24 months and were evaluated according to the Ratingen score and the total modified Sharp/van der Heijde score (SHS). RESULTS: Of 192 included patients, 166 were available for the intent-to-treat analysis (ITT). Seventy-six patients completed the study per protocol (PP). Radiographic progression (increase in the Ratingen score) was significantly less with prednisolone than with placebo. The difference in the progression rate between the groups was greatest in the first 6 months. At 24 months in the ITT population, the least squares (LS) mean difference was 3.14 (95% confidence interval [95% CI] 0.94, 5.34), P = 0.006. The results were confirmed by the total SHS in the ITT population (LS mean difference 7.20 [95% CI 0.93, 13.47], P = 0.022) and with the PP population. Clinical and functional outcomes tended to be better and the rate of remissions was higher in the prednisolone group. Side effects were observed more frequently in the prednisolone group than in the control group: weight gain (4 versus 0 patients), hypertension (6 versus 2 patients), glaucoma (3 versus 0 patients), Cushing's syndrome (5 versus 0 patients), gastric distress (9 versus 4 patients), and gastric ulcers (only with concomitant nonsteroidal antiinflammatory drug therapy; 3 versus 0 patients). No new lumbar fractures were found in either group. CONCLUSION: The very low daily dose of 5 mg prednisolone given over 2 years in combination with background DMARD therapy substantially decreased radiographic progression in early RA at low risk. PMID- 16255012 TI - Morris Ziff, PhD, MD, 1913-2005. PMID- 16255013 TI - Prednisolone in early rheumatoid arthritis: an antiinvasive effect. PMID- 16255014 TI - The need for new classification criteria for rheumatoid arthritis. PMID- 16255015 TI - Microparticles as regulators of inflammation: novel players of cellular crosstalk in the rheumatic diseases. PMID- 16255016 TI - Is radiation synovectomy for arthritis of the knee more effective than intraarticular treatment with glucocorticoids? Results of an eighteen-month, randomized, double-blind, placebo-controlled, crossover trial. AB - OBJECTIVE: To compare the clinical efficacy and safety of radiation synovectomy (RSO) with intraarticular (IA) yttrium-90 plus glucocorticoids (GCs) with the efficacy and safety of IA placebo yttrium plus GCs and to identify parameters that predict efficacy. METHODS: The knees of 97 patients with persistent arthritis despite outpatient treatment with IA GCs (n = 113 knees), were treated with either IA (90)Y plus GCs (50%) or IA placebo yttrium plus GCs (50%), followed by 3 days of bed rest in the hospital clinic, with splinting of the treated knee. Predominant diagnoses were undifferentiated arthritis (39%) and rheumatoid arthritis (32%). The clinical effect of therapy was assessed at 6 months using a composite change index (CCI; range 0-12). The primary outcome measure was the response rate (i.e., the percentage of joints with a CCI > or =6). Knees with persistent arthritis after 6 months underwent crossover therapy (51% of the (90)Y plus GCs group versus 45% of the placebo plus GCs group). Adverse effects and radiologic damage during followup were documented. RESULTS: Neither the response rate (48% in both groups), the mean CCI, nor the duration of remission was significantly different between groups. No clinically relevant short-term adverse effects were observed, except for progression of radiologic damage in 34% of the (90)Y plus GCs group versus 28% of the placebo plus GCs group (knee prosthesis placement in 8% versus 1%). The functional and radiologic status at study entry predicted the clinical effect. CONCLUSION: Treatment with (90)Y plus GCs with bed rest and splinting is not superior to IA GCs with bed rest and splinting. Over the short term, both treatments appeared to be safe, although a negative effect of (90)Y on cartilage and bone cannot be ruled out. Thus, it appears that RSO with (90)Y should no longer be considered the treatment of first choice for persistent arthritis of the knee. PMID- 16255017 TI - Infections in patients with rheumatoid arthritis treated with biologic agents. AB - OBJECTIVE: To estimate the incidence rates of serious and nonserious infections in patients with rheumatoid arthritis (RA) who start treatment with a biologic agent, and to compare these rates with those in patients with RA who receive conventional treatment. METHODS: Patients enrolled in the German biologics register between May 2001 and September 2003 were included. Treating rheumatologists assessed adverse events and serious adverse events. All adverse events and serious adverse events experienced within 12 months after study entry were analyzed. Propensity score methods were applied to estimate which part of a rate increase was likely to be attributable to differences in patient characteristics. RESULTS: Data were available for 512 patients receiving etanercept, 346 patients receiving infliximab, 70 patients receiving anakinra, and 601 control patients treated with disease-modifying antirheumatic drugs. The total number of adverse events per 100 patient-years was 22.6 (95% confidence interval [95% CI] 18.7-27.2) among patients receiving etanercept, 28.3 (95% CI 23.1-34.7) among patients receiving infliximab, and 6.8 (95% CI 5.0-9.4) among controls (P < 0.0001). Significant differences in the rate of serious adverse events were also observed. For patients receiving etanercept, those receiving infliximab, and controls, the total numbers of serious adverse events per 100 patient-years were 6.4 (95% CI 4.5-9.1), 6.2 (95% CI 4.0-9.5), and 2.3 (95% CI 1.3-3.9), respectively (P = 0.0016). After adjusting for differences in the case patient mix, the relative risks of serious adverse events were 2.2 (95% CI 0.9 5.4) for patients receiving etanercept and 2.1 (95% CI 0.8-5.5) for patients receiving infliximab, compared with controls. CONCLUSION: Patients treated with biologic agents have a higher a priori risk of infection. However, our data suggest that this risk is increased by treatment with tumor necrosis factor inhibitors. PMID- 16255018 TI - Relative contribution of cardiovascular risk factors and rheumatoid arthritis clinical manifestations to atherosclerosis. AB - OBJECTIVE: To estimate the contribution of cardiovascular (CV) risk factors and rheumatoid arthritis (RA) disease manifestations to atherosclerosis in RA. METHODS: We used high-resolution carotid ultrasound to measure the carotid intima media thickness (IMT) and plaque in 631 RA patients. Using R(2) measures from multivariable models, we estimated the contribution of demographic characteristics (age, sex, and ethnic group), CV risk factors (diabetes mellitus, hypercholesterolemia, cigarette smoking, hypertension, and body mass index, and RA manifestations (joint tenderness, swelling, and deformity, nodules, erythrocyte sedimentation rate [ESR], C-reactive protein, rheumatoid factor, the HLA-DRB1 shared epitope, and cumulative glucocorticoid dose) to each of the outcomes. Estimates were obtained in the full sample, and within strata defined by age, sex, and ethnic group. We tested for interaction between CV risk factors and RA manifestations. RESULTS: The contribution of demographic factors, CV risk factors, and RA manifestations to IMT and plaque R(2) varied depending on the patients' age stratum. Demographic features explained 11-16% of IMT variance, CV risk factors explained 4%-12%, and RA manifestations explained 1-6%. The greatest contribution of RA manifestations occurred in the youngest age group, while that of CV risk factors occurred in the older age groups. Results for carotid plaque were similar. There was a significant interaction between the number of CV risk factors present and the ESR, suggesting that the ESR's effect on IMT varied according to the number of CV risk factors. CONCLUSION: Both established CV risk factors and manifestations of RA inflammation contribute significantly to carotid atherosclerosis in RA, and may modify one another's effects. These findings may have implications regarding the prevention of atherosclerosis in RA. PMID- 16255019 TI - Influence of HLA-DR genes on the production of rheumatoid arthritis-specific autoantibodies to citrullinated fibrinogen. AB - OBJECTIVE: Antibodies directed against citrullinated fibrinogen are highly specific for rheumatoid arthritis (RA). This study was undertaken to test whether RA-associated HLA-DR alleles are associated with anti-citrullinated fibrinogen in RA patient sera and whether replacement of arginyl by citrullyl residues on fibrinogen peptides modifies their binding to HLA-DR molecules and their recognition by T cells. METHODS: Antikeratin, antifilaggrin, and anti citrullinated fibrinogen antibodies were assayed in RA patients who had undergone HLA-DR typing. Direct assays were performed to investigate binding of citrullinated or native fibrinogen peptides (encompassing the entire alpha- and beta-chains of fibrinogen) to purified HLA-DR molecules. T cell proliferative responses to citrullinated or native fibrinogen peptides were measured in RA patients and controls. RESULTS: HLA-DRB1*0404 was associated with anti citrullinated fibrinogen in RA sera (P = 0.002). For the RA-associated alleles HLA-DRB1*0401 and HLA-DR1, there was a nonsignificant trend toward association (P = 0.07). Multiple peptides from the alpha- and beta-chains of fibrinogen bound many HLA-DR alleles; DRB1*0404 was the best fibrinogen peptide binder. Citrullination did not influence fibrinogen peptide binding to HLA-DR or fibrinogen peptide recognition by T cells. Peripheral blood T cells that recognized native or citrullinated fibrinogen peptides were common in RA patients but not in healthy controls. CONCLUSION: The RA-associated HLA-DRB1*0404 allele is also associated with production of antibodies to citrullinated fibrinogen. DRB1*0401 and DRB1*01 tend to be associated with anti-citrullinated fibrinogen, but this is not statistically significant. Citrullination of fibrinogen peptide does not influence peptide-DR-T cell interaction. Finally, T cell proliferation in response to citrullinated or uncitrullinated fibrinogen peptides is frequent in RA patients and very infrequent in controls. PMID- 16255020 TI - Effects of antirheumatic treatments on the prostaglandin E2 biosynthetic pathway. AB - OBJECTIVE: Microsomal prostaglandin E synthase 1 (mPGES-1) is up-regulated in experimental arthritis and markedly expressed in synovial tissue biopsy samples from patients with rheumatoid arthritis (RA). This study was carried out to determine the effects of tumor necrosis factor (TNF) blockers and glucocorticoids on mPGES-1 and cyclooxygenase (COX) expression, as well as biosynthesis of PGE(2) in rheumatoid joints. METHODS: In vitro effects of TNF blockers and dexamethasone on the PGE(2) biosynthetic pathway were examined in RA synovial fluid mononuclear cells (SFMCs) by flow cytometry. PGE(2) levels in culture supernatants were measured by enzyme immunoassay. Expression of enzymes responsible for PGE(2) synthesis ex vivo was evaluated by immunohistochemistry in synovial biopsy samples obtained from 18 patients before and after treatment with TNF blockers and from 16 patients before and after intraarticular treatment with glucocorticoids. Double immunofluorescence was performed using antibodies against mPGES-1, COX-1, COX-2, and CD163. RESULTS: Double immunofluorescence revealed that mPGES-1 and COX-2 were colocalized in SFMCs as well as in RA synovial tissue cells. The addition of either TNF blockers or dexamethasone suppressed lipopolysaccharide-induced mPGES-1 and COX-2 expression in synovial fluid monocyte/macrophages in vitro and decreased the production of PGE(2). Intraarticular treatment with glucocorticoids significantly reduced both mPGES-1 and COX-2 expression in arthritic synovial tissue ex vivo. The number of COX-1 expressing cells in synovial tissue was also significantly decreased by glucocorticoid treatment. In contrast, neither mPGES-1 nor COX-2 expression in synovial tissue was significantly suppressed by anti-TNF therapy. CONCLUSION: These data are the first to demonstrate the effects of antirheumatic treatments on mPGES-1 expression in RA and suggest that the inhibition of PGE(2) biosynthesis, preferably by targeting mPGES-1, might complement anti-TNF treatment for optimal antiinflammatory results in RA. PMID- 16255021 TI - Refining the complex rheumatoid arthritis phenotype based on specificity of the HLA-DRB1 shared epitope for antibodies to citrullinated proteins. AB - OBJECTIVE: The main genetic risk factor for rheumatoid arthritis (RA), the HLA region, has been known for 25 years. Previous research has demonstrated, within the RA population, an association between HLA-DRB1 alleles carrying the shared epitope (SE) and antibodies directed against cyclic citrullinated peptides (anti CCP antibodies). We undertook this study to make the first comparison of SE allele frequencies in the healthy population with those in RA patients who do or do not harbor anti-CCP antibodies. METHODS: HLA-DRB1 typing was performed in 408 RA patients from the Leiden Early Arthritis Clinic (the Leiden EAC; a Dutch population-based inception cohort in which disease course was followed up over time), in 423 healthy Dutch controls, and in 720 affected members of 341 US multiplex (sibpair) families of Caucasian origin from the North American RA Consortium (NARAC) with well-established disease and fulfilling the American College of Rheumatology classification criteria for RA. The presence of anti-CCP antibodies was determined by enzyme-linked immunosorbent assay. RESULTS: For the Leiden EAC, the odds ratio (OR) describing the association of 2 copies of the SE allele with anti-CCP positivity (using no copies of the SE allele in the healthy control group as the referent) was 11.79 (P < 0.0001), while the OR for 1 SE allele was 4.37 (P < 0.0001). No association with the SE was observed in the Dutch anti-CCP-negative RA patients. For the NARAC families, linkage and association analysis revealed the SE to be associated only with anti-CCP-positive disease and not with anti-CCP-negative disease. Stratified analyses indicated that anti-CCP antibodies primarily mediated association of the SE with joint damage or disease persistence. CONCLUSION: HLA-DRB1 alleles encoding the SE are specific for disease characterized by antibodies to citrullinated peptides, indicating that these alleles do not associate with RA as such, but rather with a particular phenotype. PMID- 16255022 TI - Hip abduction moment and protection against medial tibiofemoral osteoarthritis progression. AB - OBJECTIVE: To test the hypothesis that a greater peak internal hip abduction moment is associated with a reduced likelihood of ipsilateral medial tibiofemoral osteoarthritis (OA) progression. METHODS: Fifty-seven persons with knee OA (by definite osteophyte presence and symptoms) were evaluated. Baseline assessments included kinematic and kinetic gait parameters, obtained with an optoelectronic camera system and force platform, with inverse dynamics used to calculate 3 dimensional moments at the joints; pain, using a separate visual analog scale for each knee; and alignment, using full-limb radiographs. Radiographs of the knee in a semiflexed position, with fluoroscopic confirmation of tibial rim alignment, were obtained at baseline and 18 months later. Disease progression was defined as worsening of the grade of medial joint space narrowing. Logistic regression obtained with generalized estimating equations was used to estimate odds ratios (ORs) for progression per unit of hip abduction moment, after excluding knees with the worst joint space grade at baseline (which could not progress). RESULTS: The 57 participants (63% women) with mild to moderate OA had a mean age of 67 years and a mean body mass index of 29. A greater internal hip abduction moment during gait was associated with a reduced likelihood of medial tibiofemoral OA progression, with OR/unit hip abduction moment of 0.52 and a 95% confidence interval (95% CI) of 0.32-0.85. This protective effect persisted after adjustment for age, sex, walking speed, knee pain severity, physical activity, varus malalignment severity, hip OA presence, and hip OA symptom presence, with an adjusted OR of 0.43 a 95% CI of 0.22-0.81. CONCLUSION: A greater hip abduction moment during gait at baseline protected against ipsilateral medial OA progression from baseline to 18 months. The likelihood of medial tibiofemoral OA progression was reduced 50% per 1 unit of hip abduction moment. PMID- 16255023 TI - Does hand osteoarthritis predict future hip or knee osteoarthritis? AB - OBJECTIVE: To evaluate the risk of future hip or knee osteoarthritis (OA) in subjects with hand OA at baseline and to evaluate whether the concurrent presence of hand OA, other risk factors for OA, or an OA biomarker (type II collagen C telopeptide degradation product [CTX-II]) further increases the risk. METHODS: Radiographs of the hands (baseline) and the hips and knees (baseline and 6.6 years later) were obtained in a randomly selected subset of participants in the Rotterdam Study who were ages 55 years and older. Radiographs were scored for the presence of OA using the Kellgren/Lawrence (K/L) system. A total of 1,235 subjects without OA of the hip/knee (K/L score 0-1) at baseline were included in the study. CTX-II levels were measured at baseline. The independent risk of future hip/knee OA in subjects with hand OA at baseline was assessed by logistic regression, as stratified for age, sex, body mass index, family history of OA, and heavy workload. RESULTS: Overall 12.1% of the participants (19.7% of those with hand OA versus 10.0% of those without) developed hip or knee OA (odds ratio [OR] 2.1 [95% confidence interval (95% CI) 1.3-3.1]). Subjects with hand OA had an increased risk of future hip OA (OR 3.0 [95% CI 1.6-5.4]), which was further increased in those with a family history of OA. Subjects with hand OA had an OR of 1.6 [95% CI 1.0-2.8) for the future development of knee OA, which was further increased in those who were overweight. Concurrent hand OA and high levels of CTX II further increased the risk of having hip or knee OA at followup (OR 4.2 [95% CI 2.3-7.8]). CONCLUSION: The presence of hand OA at baseline showed an increased risk of future hip/knee OA (higher for hip OA than for knee OA). The concurrent presence of hand OA and other OA risk factors or high CTX-II levels further increased the risk of future hip/knee OA. PMID- 16255024 TI - Delayed gadolinium-enhanced magnetic resonance imaging of cartilage in knee osteoarthritis: findings at different radiographic stages of disease and relationship to malalignment. AB - OBJECTIVE: Delayed gadolinium-enhanced magnetic resonance imaging (MRI) of cartilage (dGEMRIC) is used to examine the distribution of glycosaminoglycan in cartilage. This study sought to characterize dGEMRIC in the evaluation of knee osteoarthritis (OA) according to various radiographically determined disease parameters, and to examine the relationship between alignment of the knee joint and the lateral:medial dGEMRIC ratio. METHODS: Thirty-one patients with knee OA underwent MRI with a dGEMRIC protocol at 1.5T. Semiflexed knee radiographs and full-limb radiographs were also obtained for assessment of alignment. RESULTS: Compartments of the knee joint without joint space narrowing had a higher dGEMRIC index than those with any level of narrowing (mean 408 msec versus 365 msec; P = 0.001). In knees with 1 unnarrowed (spared) and 1 narrowed (diseased) compartment, the dGEMRIC index was greater in the spared versus the diseased compartment (mean 395 msec versus 369 msec; P = 0.001). In spared compartments, there was a trend toward a lower dGEMRIC index with increasing Kellgren/Lawrence (K/L) radiographic severity grade; the spared compartments of knees with a K/L grade 2 had a higher dGEMRIC index than those of knees with a K/L grade 4 (mean 425 msec versus 371 msec; P < 0.05). There was a range of dGEMRIC values in the spared compartments within a given K/L grade, demonstrating biochemical differentiation of disease in radiographically comparable compartments. Almost all compartments with narrowing had dGEMRIC indices of <400 msec. Valgus-aligned knees tended to have lower dGEMRIC values laterally, and varus-aligned knees tended to have lower dGEMRIC values medially; as a continuous variable, alignment correlated with the lateral:medial dGEMRIC ratio (Pearson's R = 0.43, P = 0.02). CONCLUSION: The biochemical information provided by dGEMRIC scans may augment radiography by improving the differentiation of disease status within a given radiographic grade, especially in early OA. PMID- 16255025 TI - Assessing bone loss on radiographs of the knee in osteoarthritis: a cross sectional study. AB - OBJECTIVE: The only established system to grade subchondral bone attrition in knee osteoarthritis (OA) has low interobserver reliability. In this study, our aim was to convert this system into a reliable tool for the assessment of subchondral bone loss in knee OA. METHODS: Templates that were designed to outline the normal contours of the knee were overlaid onto conventional radiographs of a random sample of 100 knees of OA patients who were awaiting total knee replacement (TKR). Seventy-five films from individuals with chronic knee pain who were not awaiting TKR and 75 films from asymptomatic control subjects were also assessed. Bone loss was graded from 0 (no attrition) to 3 (severe attrition of >10 mm); other established radiologic features were also graded. Spearman's rho was used to determine the correlation of attrition scores with other features, and logistic regression was used to explore whether definite bone attrition was associated with night pain. RESULTS: The inter- and intraobserver reliability values were high for attrition scores and for the presence of definite attrition (score > or =2). Bone attrition was evident in 62% of films from patients awaiting TKR, in 9% of films from individuals with chronic knee pain who were not awaiting TKR, and in 1% of films from controls. In all groups, the correlation between attrition and other features was weak to moderate. There was a nonsignificant association between definite bone attrition and night pain. CONCLUSION: Bone attrition is an additional dimension of knee OA that can be measured reliably. Definite attrition may be associated with night pain. PMID- 16255026 TI - Wnt-1-inducible signaling pathway protein 3 and susceptibility to juvenile idiopathic arthritis. AB - OBJECTIVE: To determine whether Wnt-1-inducible signaling pathway protein 3 (WISP3) polymorphisms are associated with susceptibility to juvenile idiopathic arthritis (JIA). METHODS: The exons and the intron/exon boundaries of the WISP3 gene were mutation-screened by denaturing high-performance liquid chromatography in 86 patients with polyarticular-course JIA (>/=5 joints affected) and 15 controls. Seven single-nucleotide polymorphisms (SNPs) were genotyped, using allelic discrimination, in a case-control study. Initially, 159 patients with polyarticular-course JIA and 263 controls were studied, followed by study of a replication cohort of 181 patients with polyarticular-course JIA and 355 controls. Available parents of patients with polyarticular-course JIA were also genotyped. Finally, other JIA subgroups were studied (initial cohort, n = 218; replication cohort, n = 213). Single-point and haplotype analysis was carried out. RESULTS: Positive association with SNP WISP3*G84A was observed and replicated in 2 cohorts of patients with polyarticular-course JIA. Specifically, homozygosity of the mutant allele (WISP3*84AA) conferred a 2-fold increased risk of disease susceptibility (for the initial cohort, odds ratio [OR] 2.1, 95% confidence interval [95% CI] 1.1-4.2, P = 0.03; for the replication cohort, OR 2.0, 95% CI 1.0-4.3, P = 0.05). Strong linkage disequilibrium was observed between SNPs; however, no haplotypic effect of an order of magnitude greater than the single-point WISP3*G84A association was observed. Using the transmission disequilibrium test, a trend toward overtransmission of the WISP3*84A allele was observed in patients with polyarticular-course JIA. No association of any WISP3 polymorphism was observed in the other JIA subgroups. CONCLUSION: Association and replication of a polymorphism within the first intron of the WISP3 gene have been shown in patients with polyarticular-course JIA. The functional significance of the WISP3*G84A SNP is being determined. PMID- 16255027 TI - Morphological impairments in retinal neurons of the scotopic visual pathway in a monkey model of Parkinson's disease. AB - Physiological abnormalities resulting from death of dopaminergic neurons of the central nervous system in Parkinson's disease also extend to the retina, resulting in impaired visual functions. In both parkinsonian patients and animal models, low levels of dopamine and loss of dopaminergic cells in the retina have been reported. However, the morphology and connectivity of their postsynaptic neurons, the amacrine cells, have not been analyzed. Here we report, with macaques chronically treated with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) as a model of Parkinson's disease, that morphological impairments in dopaminergic retinal neurons and their plexus in the inner retina are accompanied by an immunoreactivity decrease in gamma-aminobutyric acidergic and glycinergic amacrine cells. Especially deteriorated were AII amacrine cells, the main neuronal subtype postsynaptic to dopaminergic cells, which exhibited a marked loss of lobular appendages and dendritic processes. Concomitantly, electrical synapses among AII cells, as well as chemical synapses between these and rod bipolar cells, were highly deteriorated in parkinsonian monkeys. These results highlight that the scotopic visual pathway is severely impaired in the parkinsonian condition and provide a morphological basis for a number of abnormalities found in electrophysiological and psychophysical trials in Parkinson's disease patients and animal models. PMID- 16255028 TI - Immunohistochemical localization of GAD67-expressing neurons and processes in the rat brainstem: subregional distribution in the nucleus tractus solitarius. AB - The role of gamma-aminobutyric acid (GABA) in homeostatic control in the brainstem, in particular, in the nucleus tractus solitarius (NTS), is well established. However, to date, there is no detailed description of the distribution of GABAergic neurons within the NTS. The goal of the current study was to reexamine the efficacy of immunohistochemical localization of glutamic acid decarboxylase (GAD) protein, specifically the 67-kDa isoform (GAD67), as a marker for GABAergic neurons in the medulla and to provide a detailed map of GAD67-immunoreactive (-ir) cells within rat NTS by using a recently developed mouse monoclonal antibody. We describe a distribution of GAD67-ir cells in the medulla similar to that reported previously from in situ hybridization study. GAD67-ir cells were localized in regions known to contain high GABA content, including the ventrolateral medulla, raphe nuclei, and area postrema, but were absent from all motor nuclei, although dense terminal labeling was discerned in these regions. In the NTS, GAD67-ir was localized in all subregions. Semiquantitative analysis of the GAD67-ir distribution in the NTS revealed greater numbers of GAD67-ir cells medial to the solitary tract. Finally, dense GAD67 terminal labeling was found in the medial, central, intermediate, commissural, and subpostremal subregions, whereas sparse labeling was observed in the ventral subregion. Our findings support the use of immunohistochemistry for GAD67 as a marker for the localization of GABAergic cells and terminal processes in the rat brainstem. Furthermore, the reported heterogeneous distribution of GAD67-ir in the NTS suggests differential inhibitory modulation of sensory processing. PMID- 16255029 TI - Postnatal phenotype and localization of spinal cord V1 derived interneurons. AB - Developmental studies identified four classes (V0, V1, V2, V3) of embryonic interneurons in the ventral spinal cord. Very little is known, however, about their adult phenotypes. Therefore, we characterized the location, neurotransmitter phenotype, calcium-buffering protein expression, and axon distributions of V1-derived neurons in the adult mouse spinal cord. In the mature (P20 and older) spinal cord, most V1-derived neurons are located in lateral LVII and in LIX, few in medial LVII, and none in LVIII. Approximately 40% express calbindin and/or parvalbumin, while few express calretinin. Of seven groups of ventral interneurons identified according to calcium-buffering protein expression, two groups (1 and 4) correspond with V1-derived neurons. Group 1 are Renshaw cells and intensely express calbindin and coexpress parvalbumin and calretinin. They represent 9% of the V1 population. Group 4 express only parvalbumin and represent 27% of V1-derived neurons. V1-derived Group 4 neurons receive contacts from primary sensory afferents and are therefore proprioceptive interneurons. The most ventral neurons in this group receive convergent calbindin IR Renshaw cell inputs. This subgroup resembles Ia inhibitory interneurons (IaINs) and represents 13% of V1-derived neurons. Adult V1-interneuron axons target LIX and LVII and some enter the deep dorsal horn. V1 axons do not cross the midline. V1-derived axonal varicosities were mostly (>80%) glycinergic and a third were GABAergic. None were glutamatergic or cholinergic. In summary, V1 interneurons develop into ipsilaterally projecting, inhibitory interneurons that include Renshaw cells, Ia inhibitory interneurons, and other unidentified proprioceptive interneurons. PMID- 16255030 TI - Synaptic specializations exist between enteric motor nerves and interstitial cells of Cajal in the murine stomach. AB - Autonomic neurotransmission is thought to occur via a loose association between nerve varicosities and smooth muscle cells. In the gastrointestinal tract ultrastructural studies have demonstrated close apposition between enteric nerves and intramuscular interstitial cells of Cajal (ICC-IM) in the stomach and colon and ICC in the deep muscular plexus (ICC-DMP) of the small intestine. In the absence of ICC-IM, postjunctional neural responses are compromised. Although membrane specializations between nerves and ICC-IM have been reported, the molecular identity of these specializations has not been studied. Here we have characterized the expression and distribution of synapse-associated proteins between nerve terminals and ICC-IM in the murine stomach. Transcripts for the presynaptic proteins synaptotagmin, syntaxin, and SNAP-25 were detected. Synaptotagmin and SNAP-25-immunopositive nerve varicosities were concentrated in varicose regions of motor nerves and were closely apposed to ICC-IM but not smooth muscle. W/W(V) mice were used to examine the expression and distribution of synaptic proteins in the absence of ICC-IM. Transcripts encoding synaptotagmin, syntaxin, and SNAP-25 were detected in W/W(V) tissues. In the absence of ICC-IM, synaptotagmin and SNAP-25 were localized to nerve varicosities. Reverse transcriptase polymer chain reaction (RT-PCR) and immunohistochemistry demonstrated the expression of postsynaptic density proteins PSD-93 and PSD-95 in the stomach and expression levels of PSD-93 and PSD-95 were reduced in W/W(V) mutants. These data support the existence of synaptic specializations between enteric nerves and ICC-IM in gastric tissues. In the absence of ICC-IM, components of the synaptic vesicle docking and fusion machinery is trafficked and concentrated in enteric nerve terminals. PMID- 16255032 TI - Neuronal architecture of the mosquito deutocerebrum. AB - Mosquito behavior is heavily dependent on olfactory and mechanosensory cues, which are detected by receptor neurons on the antenna and on the palps. Recent progress in mosquito sensory genomics highlights the need for an up-to-date understanding of the neural architecture of the mosquito brain. Here we present a detailed description of the neural structure of the primary target of the majority of these neurons, the deutocerebrum, in the African malaria (Anopheles gambiae) and yellow fever (Aedes aegypti) mosquitoes. Special focus is made on the olfactory system, the antennal lobe (AL), where we present high-resolution three-dimensional models of the ALs of male and female Ae. aegypti. These models reveal a sexual dimorphism in the number of glomeruli, 49 and 50 glomeruli in male and female mosquitoes, respectively, and in the size of several of the identified glomeruli. The fine structure of receptor neuron terminations in the AL and the rest of the deutocerebrum is described, as are the arborizations of intrinsic deutocerebral neurons and neurons providing output to higher brain areas. In the AL a specific and very large center receiving input from the mechanosensory Johnston's organ is revealed as a multilobed structure receiving peripheral input according to a somatotopic pattern. Within the antennal nerve a specific neuropil containing early, bouton-like ramifications of receptor neurons is described. Within the glomerular array of the AL, neurons providing a possible feedback circuit to antennal receptor neurons are shown. With these results we provide a new resolution in mosquito deutocerebral architecture. PMID- 16255031 TI - Comparison of alpha2 nicotinic acetylcholine receptor subunit mRNA expression in the central nervous system of rats and mice. AB - The nicotinic acetylcholine receptor (nAChR) alpha2 subunit was the first neuronal nAChR to be cloned. However, data for the distribution of alpha2 mRNA in the rodent exists in only a few studies. Therefore, we investigated the expression of alpha2 mRNA in the rat and mouse central nervous systems using nonradioactive in situ hybridization histochemistry. We detected strong hybridization signals in cell bodies located in the internal plexiform layer of the olfactory bulb, the interpeduncular nucleus of the midbrain, the ventral and dorsal tegmental nuclei, the median raphe nucleus of the pons, the ventral part of the medullary reticular nucleus, the ventral horn in the spinal cord of both rats and mice, and in a few Purkinje cells of rats, but not of mice. Cells that moderately express alpha2 mRNA were localized to the cerebral cortex layers V and VI, the subiculum, the oriens layer of CA1, the medial septum, the diagonal band complex, the substantia innominata, and the amygdala of both animals. They were also located in a few midbrain nuclei of rats, whereas in mice they were either few or absent in these areas. However, in the upper medulla oblongata alpha2 mRNA was expressed in several large neurons of the gigantocellular reticular nucleus and the raphe magnus nucleus of mice, but not of rats. The data obtained show that a similar pattern of alpha2 mRNA expression exists in both rats and mice, with the exception of a few regions, and provide the basis for cellular level analysis. PMID- 16255033 TI - Dual, multilayered somatosensory maps formed by antennal tactile and contact chemosensory afferents in an insect brain. AB - The antennae of most insects move actively and detect the physical and chemical composition of objects encountered by using their associated tactile sensors. Positional information is required for these sensory modalities to interpret the physical environment. Although we have a good understanding of antennal olfactory pathways, little is known about the destinations of antennal mechanosensory and contact chemosensory (gustatory) receptor neurons in the central nervous system. The cockroach Periplaneta is equipped with a pair of long, thin antennae, which are covered in bristles. The distal portions of each antenna possess about 6,500 bimodal bristles that house one tactile sensory and one to four contact chemosensory neurons. In this study, we investigated the morphologies of bimodal bristle receptor afferents by staining individual or populations of bristles. Unlike olfactory afferents, which project exclusively into the glomeruli in the ventral region of the deutocerebrum, both the presumptive mechanosensory and the contact chemosensory afferents projected into the posterior dorsal region of the deutocerebrum and the anterior region of the subesophageal ganglion. Each afferent showed multilayered segmentation and spatial occupation reflecting its three-dimensional position in the periphery. Presumptive contact chemosensory afferents, characterized by their thin axons and unique branching pattern, occupied more medioventral positions compared with the presumptive tactile afferents. Furthermore, projection fields of presumptive contact chemosensory afferents from single sensilla tended to be segregated from each other. These observations suggest that touch and taste positional information from the antenna is precisely represented in primary centers in a modality-specific manner. PMID- 16255034 TI - Expression and function of the neuronal gap junction protein connexin 36 in developing mammalian retina. AB - With the advent of transgenic mice, much has been learned about the expression and function of gap junctions. Previously, we reported that retinal ganglion cells in mice lacking the neuronal gap junction protein connexin 36 (Cx36) have nearly normal firing patterns at postnatal day 4 (P4) but many more asynchronous action potentials than wild-type mice at P10 (Torborg et al. [2005] Nat. Neurosci. 8:72-78). With the goal of understanding the origin of this increased activity in Cx36-/- mice, we used a transgenic mouse (Deans et al. [2001] Neuron 31:477-485) to characterize the developmental expression of a Cx36 reporter in the retina. We found that Cx36 was first detected weakly at P2 and gradually increased in expression until it reached an adult pattern at P14. Although the onset of expression varied by cell type, we identified Cx36 in the glycinergic AII amacrine cell, glutamatergic cone bipolar cell, and retinal ganglion cells (RGCs). In addition, we used calcium imaging and multielectrode array recording to characterize further the firing patterns in Cx36-/- mice. Both correlated and asynchronous action potentials in P10 Cx36-/- RGCs were significantly inhibited by bath application of an ionotropic glutamate receptor antagonist, indicating that the increase in activity was synaptically mediated. Hence, both the expression patterns and the physiology suggest an increasing role for Cx36 containing gap junctions in suppressing RGC firing between waves during postnatal retinal development. PMID- 16255035 TI - Nitrate promotes biological oxidation of sulfide in wastewaters: experiment at plant-scale. AB - Biogenic production of sulfide in wastewater treatment plants involves odors, toxicity and corrosion problems. The production of sulfide is a consequence of bacterial activity, mainly sulfate-reducing bacteria (SRB). To prevent this production, the efficiency of nitrate addition to wastewater was tested at plant scale by dosing concentrated calcium nitrate (Nutriox) in the works inlet. Nutriox dosing resulted in a sharp decrease of sulfide, both in the air and in the bulk water, reaching maximum decreases of 98.7% and 94.7%, respectively. Quantitative molecular microbiology techniques indicated that the involved mechanism is the development of the nitrate-reducing, sulfide-oxidizing bacterium Thiomicrospira denitrificans instead of the direct inhibition of the SRB community. Denitrification rate in primary sedimentation tanks was enhanced by nitrate, being this almost completely consumed. No significant increase of inorganic nitrogen was found in the discharged effluent, thus reducing potential environmental hazards to receiving waters. This study demonstrates the effectiveness of nitrate addition in controlling sulfide generation at plant scale, provides the mechanism and supports the environmental adequacy of this strategy. PMID- 16255037 TI - Purification of monoclonal antibodies derived from transgenic goat milk by ultrafiltration. AB - With the goal of recovering heterologous immunoglobulin (IgG), which comprises 10 15% of the total proteins, from transgenic goat milk at 80% yield and 80% purity, we have developed and tested a two-step membrane isolation and purification process. In the first step, reported earlier by Baruah and Belfort, microfiltration was used to fractionate the milk proteins and recover > 90% of the original IgG at a purity of about 15-20% in the permeate stream. Here, we focus on ultrafiltration (UF) to increase the purity of the target protein to 80%, while maintaining a relatively high IgG yield (80%). Tangential flow UF experiments in diafiltration mode were conducted with 100 kDa cellulosic membranes to evaluate the optimal pH, ionic strength, and uniform transmembrane pressure (TMP). The TMP was kept uniform by permeate circulation in co-flow mode. The traditional approach of conducting the UF process close to the pI of the predominant whey proteins (15-40 kDa, pI 5.2), to transmit these proteins while retaining heterologous IgG (155 kDa), could not be applied here because of precipitation of residual casein at pH values lower than 8.5. Instead, the packing characteristics of the cake layer on the membrane wall, as elucidated in the Aggregate Transport Model presented by Baruah et al. was utilized to achieve a selectivity of > 15, which was sufficient to meet the stated goals of purity and yield for this difficult separation. This combined process is expected to reduce the load on subsequent purification and polishing steps for eventual therapeutic use. PMID- 16255036 TI - A novel mechanism of neutrophil recruitment in a coculture model of the rheumatoid synovium. AB - OBJECTIVE: Rheumatoid arthritis (RA) is classically thought of as a Th1, T lymphocyte-driven disease of the adaptive immune system. However, cells of the innate immune system, including neutrophils, are prevalent within the diseased joint, and accumulate in large numbers. This study was undertaken to determine whether cells of the rheumatoid stromal microenvironment could establish an inflammatory environment in which endothelial cells are conditioned in a disease specific manner to support neutrophil recruitment. METHODS: Human umbilical vein endothelial cells (ECs) and fibroblasts isolated from the synovium or skin of RA patients were established in coculture on opposite sides of porous transwell filters. After 24 hours of EC conditioning, the membranes were incorporated into a parallel-plate, flow-based adhesion assay and levels of neutrophil adhesion to ECs were measured. RESULTS: ECs cocultured with synovial, but not skin, fibroblasts could recruit neutrophils in a manner that was dependent on the number of fibroblasts. Antibody blockade of P-selectin or E-selectin reduced neutrophil adhesion, and an antibody against CD18 (the beta2 integrin) abolished adhesion. Blockade of CXCR2, but not CXCR1, also greatly inhibited neutrophil recruitment. Interleukin-6 (IL-6) was detectable in coculture supernatants, and both IL-6 and neutrophil adhesion were reduced in a dose-dependent manner by hydrocortisone added to cocultures. Antibody blockade of IL-6 also effectively abolished neutrophil adhesion. CONCLUSION: Synovial fibroblasts from the rheumatoid joint play an important role in regulating the recruitment of inflammatory leukocytes during active disease. This process may depend on a previously unsuspected route of IL-6-mediated crosstalk between fibroblasts and endothelial cells. PMID- 16255038 TI - Tailor-made olefinic medium-chain-length poly[(R)-3-hydroxyalkanoates] by Pseudomonas putida GPo1: batch versus chemostat production. AB - Functionalized medium-chain-length polyhydroxyalkanoates (mclPHAs) have gained much interest in research on biopolymers because of their ease of chemical modification. Tailored olefinic mclPHA production from mixtures of octanoic acid and 10-undecenoic acid was investigated in batch and dual (C,N) nutrient limited chemostat cultures of Pseudomonas putida GPo1 (ATCC 29347). In a batch culture, where P. putida GPo1 was grown on a mixture of octanoic acid (58 mol%) and 10 undecenoic acid (42 mol%), it was found that the fraction of aliphatic monomers was slightly lower in mclPHA produced during exponential growth than during late stationary phase. Thus, the total monomeric composition changed over time indicating different kinetics for the two carbon substrates. Chemostat experiments showed that the dual (C,N) nutrient limited growth regime (DNLGR) for 10-undecenoic acid coincided with the one for octanoic acid. Five different chemostats on equimolar mixtures of octanoic acid and 10-undecenoic acid within the DNLGR revealed that the monomeric composition of mclPHA was not a function of the carbon to nitrogen (C(0)/N(0)) ratio in the feed medium but rather of the dilution rate. The fraction of aliphatic monomers in the accumulated mclPHA was slightly lower at high dilution rates and increased towards low dilution rates, again indicating different kinetics for the two carbon substrates in P. putida GPo1. PMID- 16255040 TI - Determination of the baculovirus transducing titer in mammalian cells. AB - Baculovirus has emerged as a promising vector for in vivo or ex vivo gene therapy. To date, the infectious titer and multiplicity of infection (MOI) based on the ability of baculovirus to infect insect cells are commonly adopted to indicate the virus dosage. However, the infectious titer and MOI do not reliably represent the baculovirus transducing ability, making the comparison of baculovirus-mediated gene transfer difficult. To determine the baculovirus transducing ability more rapidly and reliably, we developed a protocol to evaluate the transducing titers of baculovirus stocks. The virus was diluted twofold serially and used to transduce HeLa cells. The resultant transduction efficiencies were measured by flow cytometry for the calculation of transducing titers. Compared to the infectious titer, the determination of transducing titer is more reproducible as the standard deviations among measurements are smaller. Also, the transducing titers can be obtained in 24 h, which is significantly faster as opposed to 4-7 days to obtain the infectious titer. More importantly, we demonstrated that baculoviruses with higher transducing titers could transduce cells at higher efficiency and yield stronger and longer transgene expression, confirming that the transducing titer was representative of the baculovirus transducing ability. This finding is particularly significant for ex vivo gene delivery whereby unconcentrated viruses are used for transduction and long-term transgene expression is desired. In this regard, our titration protocol provides a simple, fast, and reliable measure to evaluate the quality of virus stocks during virus production and purification, and is helpful to predict the performance of vector supernatants and ensure reproducible gene delivery experiments. PMID- 16255039 TI - Extracellular superoxide dismutase and oxidant damage in osteoarthritis. AB - OBJECTIVE: To use human cartilage samples and a mouse model of osteoarthritis (OA) to determine whether extracellular superoxide dismutase (EC-SOD) is a constituent of cartilage and to evaluate whether there is a relationship between EC-SOD deficiency and OA. METHODS: Samples of human cartilage were obtained from femoral heads at the time of joint replacement surgery for OA or femoral neck fracture. Samples of mouse tibial cartilage obtained from STR/ort mice and CBA control mice were compared at 5, 15, and 35 weeks of age. EC-SOD was measured by enzyme-linked immunosorbent assay, Western blotting, and immunohistochemistry techniques. Real-time quantitative reverse transcription-polymerase chain reaction was used to measure messenger RNA for EC-SOD and for endothelial cell, neuronal, and inducible nitric oxide synthases. Nitrotyrosine formation was assayed by Western blotting in mouse cartilage and by fluorescence immunohistochemistry in human cartilage. RESULTS: Human articular cartilage contained large amounts of EC-SOD (mean +/- SEM 18.8 +/- 3.8 ng/gm wet weight of cartilage). Cartilage from patients with OA had an approximately 4-fold lower level of EC-SOD compared with cartilage from patients with hip fracture. Young STR/ort mice had decreased levels of EC-SOD in tibial cartilage before histologic evidence of disease occurred, as well as significantly more nitrotyrosine formation at all ages studied. CONCLUSION: EC-SOD, the major scavenger of reactive oxygen species in extracellular spaces, is decreased in humans with OA and in an animal model of OA. Our findings suggest that inadequate control of reactive oxygen species plays a role in the pathophysiology of OA. PMID- 16255041 TI - Macroscopic and microscopic features of synovial membrane inflammation in the osteoarthritic knee: correlating magnetic resonance imaging findings with disease severity. AB - OBJECTIVE: To determine the magnetic resonance imaging (MRI), macroscopic, and microscopic characteristics of synovial membrane inflammation, to study the relationship between disease severity and the degree of synovial inflammation on MRI and on macroscopic and microscopic examination, and to look for colocalization of chondral lesions and synovial inflammation. METHODS: Thirty nine patients with knee osteoarthritis (OA) were classified into 2 groups according to the severity of cartilage lesions as revealed by chondroscopy. Group 1 (n = 14) had mild cartilage lesion(s) without exposure of subchondral bone. Group 2 (n = 25) had severe cartilage lesion(s) with focal or diffuse exposure of subchondral bone. Synovitis was evaluated on T1-weighted MRI sequences according to the degree of synovial thickening on a 4-point scale (ranging from 0 to 3) in 5 regions of interest. Synovial membrane was macroscopically scored, and biopsies were performed on the 5 preselected sites for histologic scoring. RESULTS: The mean +/- SD synovial thickening score on MRI was 1.55 +/- 0.90, with no significant difference between groups 1 and 2. Intra- and interobserver reproducibility of the total synovial score was excellent, and interobserver reproducibility of the MRI grade was good. Synovitis was diffuse and associated with chondral lesions only in the medial femorotibial compartment (r = 0.49, P = 0.001). The degree of synovial thickening on MRI correlated with qualitative macroscopic analysis (r(s) = 0.58, P < 0.001) and with microscopic features (synovial lining cells [r(s) = 0.23, P < 0.007], surface fibrin deposition [r(s) = 0.12, P < 0.01], fibrosis [r(s) = 0.31, P < 0.006], edema [r(s) = 0.17, P = 0.07], congestion [r(s) = 0.30, P < 0.005], and infiltration [r(s) = 0.46, P < 0.0001]). Fibrin and infiltration parameters were more severe in end-stage disease (P = 0.009 and P = 0.02, respectively). CONCLUSION: Synovitis may be present from the onset of OA and may be evaluated on MRI. MRI evaluation of synovitis could be used to classify OA patients in clinical trials and could help to identify those who could benefit from synovium-targeted therapy. PMID- 16255042 TI - Investigating the aspartic acid (D) repeat of asporin as a risk factor for osteoarthritis in a UK Caucasian population. AB - OBJECTIVE: A compelling genetic association with osteoarthritis (OA) of 2 functional alleles in the aspartic acid (D) repeat of the asporin gene was recently reported in a Japanese population. Allele D13 of the repeat encoded OA protection, whereas allele D14 encoded OA susceptibility. The 2 alleles mediate differences in the capacity of asporin to inhibit the cartilage growth factor transforming growth factor beta, with the D14 allele being a particularly potent inhibitor. Our objective was to assess whether the D repeat is associated with OA in UK Caucasians. METHODS: The repeat was genotyped in 1,247 patients who had undergone elective joint replacement of the hip or the knee due to end-stage primary OA and in 748 age-matched controls. RESULTS: The D13 allele was more common in controls, and the D14 allele was more common in patients. However, this trend was significant only for men who had undergone hip replacement (P = 0.016, odds ratio 1.48, 95% confidence interval 1.09-2.01). CONCLUSION: Our data suggest that the asporin polymorphism is not a major influence on OA etiology in Caucasians. The results of our study do not question the veracity of the Japanese report. Instead, our study highlights the complex, heterogeneous nature of OA genetic susceptibility. PMID- 16255043 TI - The lateral view radiograph for assessment of the tibiofemoral joint space in knee osteoarthritis: its reliability, sensitivity to change, and longitudinal validity. AB - OBJECTIVE: To evaluate the reliability, validity, and sensitivity to change of tibiofemoral (TF) narrowing on lateral radiographic views. METHODS: In a natural history study of symptomatic knee osteoarthritis (OA), both lateral view and fluoroscopically positioned posteroanterior (PA) semiflexed view radiographs of the knee in 30 degrees of flexion and with weight bearing were obtained at baseline and at 30 months. Test-retest reliability was evaluated using repeat radiographs, with joint space width measured using electronic calipers. All radiographs were scored on a 0-3 scale, and progression of joint space loss was defined as narrowing of the joint space by 1 grade. We evaluated sensitivity to change compared with the PA view. We evaluated validity by examining whether knees with progression showed expected malalignment on full-limb films. RESULTS: Test-retest reliability of the TF joint space using the lateral view had a root mean square error of 0.303 mm, with 92.5% of repeats within 1 mm. More knees showed progression on the lateral view alone (n = 41) than on the PA view alone (n = 27). Compared with knees without joint space loss, knees with medial compartment loss on the lateral view only were more varus malaligned (P < 0.001), while those with lateral compartment loss were more valgus malaligned (P = 0.008). CONCLUSION: In the assessment of TF joint space loss, lateral view radiographs are reliable, valid, and more sensitive to change than fluoroscopically positioned PA radiographs. PMID- 16255044 TI - Patterns of clinical remission in select categories of juvenile idiopathic arthritis. AB - OBJECTIVE: To characterize disease activity patterns in a large cohort of children with juvenile idiopathic arthritis (JIA), by applying newly developed preliminary definitions of inactive disease, clinical remission on medication, and clinical remission off medication. METHODS: Children with persistent or extended oligoarthritis, polyarthritis (either rheumatoid factor [RF] positive or RF negative), or systemic JIA who had been followed up for a period of at least 4 years were evaluated for episodes of inactive disease, clinical remission on medication, and clinical remission off medication. Descriptive statistics, correlation analyses, and survival analyses were performed. RESULTS: Four hundred thirty-seven children met the criteria for review. Three hundred ninety-one patients (89%) experienced a total of 878 episodes of inactive disease, with a median episode length of 12.7 months. Two hundred twenty-eight episodes of inactive disease (26%) resulted in clinical remission off medication; it was equally as likely that episodes of inactive disease would or would not follow a period of clinical remission on medication. Thirty-six percent of episodes of clinical remission off medication persisted for at least 2 years, and only 6% of such episodes persisted for 5 years. RF-positive patients were the least likely to achieve clinical remission off medication (5%), and patients with persistent oligoarticular JIA were the most likely (68%). Among patients with persistent oligoarticular JIA, most of the disease course was characterized by inactive disease; in most other patients the majority of the disease course involved active disease. CONCLUSION: Using newly developed preliminary criteria for inactive disease, clinical remission on medication, and clinical remission off medication, we observed that only one-fourth of 878 episodes of inactive disease resulted in clinical remission off medication during followup of at least 4 years. Only a small proportion of episodes of clinical remission off medication were sustained for >5 years. These results highlight the critical need for therapies that have the ability to induce sustained remission of JIA. PMID- 16255045 TI - Utility of corticosteroid injection for temporomandibular arthritis in children with juvenile idiopathic arthritis. AB - OBJECTIVE: To assess the effects of computed tomography (CT)-guided injection of corticosteroid into the temporomandibular joint (TMJ) in children with juvenile idiopathic arthritis (JIA) and clinical and magnetic resonance imaging (MRI) evidence of TMJ inflammation. METHODS: Twenty-three children ages 4-16 years with JIA and MRI evidence of TMJ inflammation received CT-guided TMJ injections of corticosteroid (triamcinolone acetonide [n = 16] or triamcinolone hexacetonide [n = 7]). Jaw pain or dysfunction and maximal incisal opening (MIO) distance were assessed before and after injection. Fourteen patients had followup MRI studies of the TMJ 6-12 months after injection. RESULTS: Of the 13 patients with symptoms of jaw pain prior to corticosteroid treatment, 10 (77%) had complete resolution of pain (P < 0.05). Prior to corticosteroid injection, MIO in all 23 patients was below age-matched normal values. After injection, the MIO was improved by at least 0.5 cm in 10 patients (43%) (P = 0.0017). Patients under 6 years of age at the time of injection showed the best response, with a postinjection MIO similar to that in age-matched controls (P = 0.2267). There was involvement of 23 TMJs in the 14 patients who had followup MRI studies; resolution of effusions was observed in 11 (48%) of the TMJs. Other than short-term facial swelling in 2 patients, there were no side effects. CONCLUSION: The majority of children with symptomatic TMJ arthritis improved after intraarticular corticosteroid injection. Approximately half the patients experienced significant improvement in MIO and TMJ effusion. These data suggest that corticosteroid injection may be a useful procedure for the prevention and treatment of morbidities associated with TMJ arthritis in JIA. PMID- 16255046 TI - The effectiveness of treating juvenile dermatomyositis with methotrexate and aggressively tapered corticosteroids. AB - OBJECTIVE: Childhood dermatomyositis (DM) is often a chronic disease, lasting many years. It has traditionally been treated with long-term corticosteroid therapy; side effects are often seen. For more than a decade, methotrexate (MTX) has been safely used for the treatment of juvenile arthritis. Here, we report use of MTX as first-line therapy for DM, along with aggressively tapered corticosteroids, in an attempt to reduce treatment-related side effects. METHODS: We studied an inception cohort of 31 children with DM who were rigorously followed up in our myositis clinic, and compared them with a control group of 22 patients with incident cases of juvenile DM who received treatment just before we instituted a policy of first-line therapy with MTX. The mean starting dosage of MTX in the study group was 15 mg/m(2)/week. RESULTS: Both groups had similar improvement in strength and physical function; however, the median time during which patients in the study group received corticosteroids was 10 months, compared with 27 months for controls (P < 0.0001). As a result, the cumulative prednisone dose in the study group was approximately half that in the control group (7,574 mg versus 15,152 mg; P = 0.0006). The study group had greater height velocity during the first year of treatment and a smaller increase in the body mass index over the first 2 years. In the control group, the relative risk of cataracts developing was 1.95 (95% confidence interval 1.05-4.17). Side effects of MTX were rarely observed. CONCLUSION: Use of MTX in conjunction with an aggressively tapered course of prednisone may be as effective as traditional long term corticosteroid therapy for children with DM, while decreasing the cumulative dose of corticosteroids. PMID- 16255047 TI - Somatic mosaicism of CIAS1 in a patient with chronic infantile neurologic, cutaneous, articular syndrome. AB - Chronic infantile neurologic, cutaneous, articular syndrome (CINCA syndrome) is a severe inflammatory disease that was recently found to be associated with mutations in CIAS1. However, CIAS1 mutations have been detected in only half of CINCA syndrome patients, and it remains unclear which genes are responsible for the syndrome in the remaining patients. We describe here a patient with CINCA syndrome who exhibited CIAS1 somatic mosaicism. We genetically analyzed the CIAS1 gene in various blood cells and the buccal mucosa of the patient. The production of interleukin-1beta (IL-1beta) by peripheral blood mononuclear cells (PBMCs) was measured by enzyme-linked immunosorbent assay, and the ability of the mutant CIAS1 gene to enhance ASC-dependent NF-kappaB activation was assessed to confirm that the mutations of CIAS1 found were responsible for the patient's clinical manifestations of the CINCA syndrome. The patient had 1 heterologous single nucleotide polymorphism, 587G>A (S196N), and 1 heterologous mutation, 1709A>G (Y570C), in exon 3 of CIAS1. The latter mutation was found to occur as somatic mosaicism. The patient's PBMCs produced a large amount of IL-1beta in the absence of stimulation, unlike those from controls or from his mother, who also bore the S196N polymorphism. In addition, the Y570C mutation (with or without the S196N polymorphism) increased the ability of CIAS1 to induce ASC-dependent NF-kappaB activation, unlike the wild-type gene or the gene bearing the S196N polymorphism alone. The findings in this patient indicate that somatic mosaicism is one reason CIAS1 mutations have not been detected in some patients with CINCA syndrome. PMID- 16255048 TI - Heterologous protein production capacity of mammalian cells cultivated as monolayers and microtissues. AB - A precise understanding of processes managing heterologous protein production in vitro and in vivo is essential for the manufacture of sophisticated biopharmaceuticals as well as for future gene therapy and tissue engineering initiatives. Capitalizing on the gravity-enforced self-assembly of monodispersed cells into coherent (multicellular) microtissues we studied heterologous protein production of microtissues and monolayers derived from cell lines and primary cells engineered/transduced for (i) constitutive, (ii) proliferation-controlled, (iii) macrolide-, or (iv) gas-inducible expression of the human placental secreted alkaline phosphatase (SEAP) and of the Bacillus stearothermophilus derived secreted alpha-amylase (SAMY). Specific productivity of cells assembled in microtissues was up to 20-fold higher than isogenic monolayer cultures. Diffusion across microtissues could be further increased by HUVEC-mediated vascularization. As well as higher specific protein productivities, microtissues were also more efficient than monolayer cultures in assembling transgenic lentiviral particles. Our results showed that mammalian cells embedded in a tissue-like three-dimensional (3D) microenvironment exhibit increased production capacity. This observation should be considered for gene therapy and tissue engineering scenarios as well as for biopharmaceutical manufacturing. PMID- 16255049 TI - Expansion and enhanced survival of natural killer cells expressing the killer immunoglobulin-like receptor KIR3DL2 in spondylarthritis. AB - OBJECTIVE: The spondylarthritides (SpA) are strongly associated with possession of HLA-B27. We hypothesized that the expression of abnormal forms of HLA-B27 in SpA may have a pathogenic role through interaction with cells bearing natural killer (NK) receptors, in particular, killer immunoglobulin-like receptor (KIR) KIR3DL2, a receptor for HLA-B27 homodimer (B27(2)). We therefore undertook the present study to determine the number and function of NK and T cells bearing KIR3DL2 in SpA. METHODS: Expression of KIR3DL2 on NK and T cells was quantified in peripheral blood (PB) from 35 patients with SpA and 5 patients with juvenile enthesitis-related arthritis (juvenile ERA); samples were compared with samples from healthy and rheumatoid arthritis (RA) controls. Paired synovial fluid (SF) was studied where available. Expression of other KIRs as well as activation, memory, and homing markers on KIR3DL2+ NK and T cells was quantified. NK cell survival was assessed using the apoptotic markers annexin V and 7 aminoactinomycin D, and cytotoxicity by (51)Cr release assay. RESULTS: In SpA, an increased number of PB and SF NK and CD4+ T cells expressed the KIR3DL2 receptor compared with controls. In ERA, KIR3DL2 expression was increased in PB and SF CD4 T cells (and SF NK cells) compared with RA controls. KIR3DL2+ NK cells had an activated phenotype, and were protected from apoptosis by culture with a cell line expressing B27(2). SpA PB mononuclear NK cells from SpA patients showed greater cytotoxicity than those from controls. CONCLUSION: KIR3DL2 expression on NK cells and CD4 lymphocytes is increased in SpA and ERA. These cells are activated and may have a pathogenic role. PMID- 16255050 TI - Evidence for common genetic control in pathways of inflammation for Crohn's disease and psoriatic arthritis. AB - OBJECTIVE: Clinical, pharmacologic, and epidemiologic evidence supports the hypothesis that common genetic pathways may underlie inflammatory diseases. In a previous study, a Crohn's disease gene, CARD15, was demonstrated to be associated with psoriatic arthritis (PsA). Recently, a functional haplotype of 2 single nucleotide polymorphisms (SNPs) mapping to the organic cation transporter (OCTN) genes, SLC22A4 and SLC22A5, was identified as a second Crohn's disease susceptibility locus. The SLC22A4 gene has also been associated with rheumatoid arthritis. This study was undertaken to further elucidate associations of PsA with Crohn's disease susceptibility genes. METHODS: Association with CARD15 and OCTN was investigated in UK Caucasian patients with PsA (n = 472) and population controls (n = 594), using 5' allelic discrimination assays (TaqMan). Two SNPs in OCTN, forming a haplotype previously associated with Crohn's disease, were also tested in patients with psoriasis (n = 218) and patients with early undifferentiated inflammatory arthritis (n = 386). Allele and estimated haplotype frequencies were compared between patients and controls. RESULTS: No association of PsA with CARD15 was detected. In contrast, a functional SNP mapping to the promoter region of SLC22A5 (rs2631367) was associated with PsA (for CC versus GG, odds ratio 1.65, 95% confidence interval 1.13-2.41, uncorrected P = 0.005). In addition, the haplotype associated with Crohn's disease was also associated with PsA (P = 0.001). No association was detected in the cohort with psoriasis alone or in the cohort with undifferentiated inflammatory arthritis. CONCLUSION: The OCTN haplotype previously associated with Crohn's disease is also associated with PsA, suggesting that these 2 diseases may share some common genetic control in pathways of inflammation. PMID- 16255052 TI - Pseudodominant inheritance of the hyperimmunoglobulinemia D with periodic fever syndrome in a mother and her two monozygotic twins. AB - Hyperimmunoglobulinemia D with periodic fever syndrome (HIDS) is a recessively inherited recurrent fever syndrome. We describe a family of 2 monozygotic twins and their mother with characteristic symptoms of HIDS, but normal levels of IgD and IgA, and with a dominant inheritance pattern. Mevalonate kinase (MK) activity was deficient in both children, and analysis of the MVK gene revealed compound heterozygosity for 2 new mutations, G25G and R277H. Being positioned adjacent to a donor splice site, the G25G mutation was shown by reverse transcription polymerase chain reaction analyses to cause aberrant splicing of the MVK messenger RNA, thus being disease-relevant. The mother, who was also symptomatic during her childhood and adolescence, was a compound heterozygote for I268T and R277H. Our findings expand the genetic and ethnic spectrum of HIDS and show that the possible presence of this disease cannot be excluded based solely on inheritance patterns. In each case in which HIDS is clinically suspected, analysis of MK activity and/or the MVK gene (especially exons 9 and 11) should be performed. PMID- 16255051 TI - MEFV analysis is of particularly weak diagnostic value for recurrent fevers in Western European Caucasian patients. AB - OBJECTIVE: Familial Mediterranean fever (FMF) is an autosomal-recessive disorder characterized by recurrent attacks of fever, with abdominal, thoracic, or articular pain. FMF is particularly common in Mediterranean populations, while other populations are rarely affected. MEFV gene analysis provides the only objective diagnostic criterion for FMF. However, the spectrum of MEFV mutations, which was first established in classically affected populations, remains insufficiently studied in other populations. The purpose of this study was to assess involvement of MEFV in the phenotype of western European Caucasian patients with a clinical diagnosis of FMF. METHODS: Mutation analysis was performed in 208 Caucasian patients from western Europe, by screening for the most common MEFV mutations in exons 2, 3, 5, and 10, and by sequencing the promoter region and the whole MEFV coding sequence in 21 of these patients. RESULTS: None of the patients carried 2 mutated alleles. Only 2 patients carried 1 mutated allele. CONCLUSION: FMF-like syndromes in western European Caucasian populations cannot be explained by MEFV mutations. These results should be helpful in avoiding laborious and costly MEFV molecular analyses that, at the population level, seem to be of poor diagnostic value in the case of western European Caucasian patients, and rather should prompt a search for other causes in those patients. PMID- 16255053 TI - Cat-scratch disease-associated arthropathy. AB - OBJECTIVE: To characterize the articular manifestations of cat-scratch disease (CSD) and to evaluate the long-term clinical outcome of those manifestations. METHODS: A community- and hospital-based surveillance study of CSD was conducted in Israel between 1991 and 2002. CSD was defined as present in a patient when a compatible clinical syndrome and a positive confirmatory finding of Bartonella henselae (by serology and/or polymerase chain reaction) were identified. CSD patients with arthropathy (arthritis/arthralgia) that limited or precluded usual activities of daily living constituted the study group. Patients were followed up until > or =6 weeks after resolution of symptoms, or if symptoms persisted, for >/=12 months. CSD patients without arthropathy served as controls. RESULTS: Among 841 CSD patients, 24 (2.9%) had rheumatoid factor-negative arthropathy that was often severe and disabling. Both univariate and multivariate analyses identified female sex (67% of arthropathy patients versus 40% of controls; relative risk [RR] 2.5, P = 0.047), age older than 20 years (100% of arthropathy patients versus 43% of controls; RR 4.9, P = 0.001), and erythema nodosum (21% of arthropathy patients versus 2% of controls; RR 7.9, P = 0.001) as variables significantly associated with arthropathy. Knee, wrist, ankle, and elbow joints were most frequently affected. Ten patients (42%) had severe arthropathy in the weight-bearing joints, which substantially limited their ability to walk, and 4 of these patients were hospitalized. All of the patients had regional lymphadenopathy, 37.5% had nocturnal joint pain, and 25% had morning stiffness. Nineteen patients (79.2%) recovered after a median duration of 6 weeks (range 1 24 weeks), whereas 5 patients (20.8%) developed chronic disease persisting 16-53 months (median 30 months) after the onset of arthropathy. CONCLUSION: This is the first comprehensive study of arthropathy in CSD. CSD-associated arthropathy is an uncommon syndrome affecting mostly young and middle-age women. It is often severe and disabling, and may take a chronic course. PMID- 16255054 TI - The antiangiogenic tissue kallikrein pattern of endothelial cells in systemic sclerosis. AB - OBJECTIVE: Postnatal angiogenesis relies on a proper response of endothelial cells to angiogenic stimuli. In systemic sclerosis (SSc), endothelial cells are unresponsive to angiogenic factors. Since circumstantial and experimental evidence points to tissue kallikreins as powerful effectors of the angiogenic response, we undertook this study to investigate the kallikrein pattern of normal and SSc endothelial cells in order to identify differences that can account for defective angiogenesis. METHODS: Expression of 14 tissue kallikreins was studied by a microarray approach, by reverse transcription-polymerase chain reaction, and by Western blotting in endothelial cells isolated from the skin of clinically healthy subjects and SSc patients. Cell proliferation was quantified by direct cell counting. Invasion and capillary morphogenesis were evaluated in a Boyden chamber and in culture flasks layered with Matrigel. Cyclic nucleotide production was measured by enzyme immunoassay. MAP kinase and ERK activation were measured by Western blotting. RESULTS: Endothelial cells from SSc patients showed poor expression of kallikreins 9, 11, and 12 compared with endothelial cells from normal subjects. Antibodies against the relevant kallikreins on normal endothelial cells revealed that while kallikreins 9, 11, and 12 induced cell growth, only kallikrein 12 regulated invasion and capillary morphogenesis. Buffering of kallikrein 12 with antibodies resulted in the acquisition of an SSc like pattern by normal cells in in vitro angiogenesis. Reduction of cAMP and cGMP production and of ERK phosphorylation upon administration of antikallikrein antibodies revealed that the activity of kallikreins 9, 11, and 12 was mediated by kinins. CONCLUSION: Reduction of tissue kallikreins 9, 11, and 12 may be relevant to reduced angiogenesis in SSc patients. PMID- 16255055 TI - Inhibition of lupus disease by anti-double-stranded DNA antibodies of the IgM isotype in the (NZB x NZW)F1 mouse. AB - OBJECTIVE: In systemic lupus erythematosus (SLE), immune complexes (ICs) containing pathogenic IgG anti-double-stranded DNA (anti-dsDNA) autoantibodies are deposited in renal capillaries and initiate glomerulonephritis (GN) by the activation of complement and effector cells. In contrast, it has been demonstrated that the presence of IgM anti-dsDNA antibodies correlates negatively with the development of GN in SLE. The aim of this study was to determine whether anti-dsDNA antibodies of the IgM isotype protect against IC-mediated organ damage in SLE. METHODS: Lupus-prone (NZB x NZW)F(1) mice (females) were treated with murine monoclonal IgM anti-dsDNA antibodies. Treatment was delivered by subcutaneous injection at a dosage of 100 mug/week starting at 16 weeks of age (prophylactic) or at 24 weeks of age (therapeutic). RESULTS: Mice treated with IgM anti-dsDNA exhibited a delayed onset of proteinuria and a reduced degree of renal pathology, which resulted in significantly improved survival as compared with control mice. Serum concentrations of IgG anti-dsDNA antibodies were not significantly modified. However, glomerular deposition of ICs was markedly reduced in both treatment protocol groups. In contrast, higher amounts of IgG and IgM and increased expression of Fcgamma receptor were demonstrated in liver sections from the treated mice compared with the untreated mice, suggesting an enhanced clearance of soluble ICs from phagocytic cells of the reticuloendothelial system. CONCLUSION: These data demonstrate the efficacy of IgM anti-dsDNA treatment in inhibiting the pathologic changes of lupus in (NZB x NZW)F(1) mice. Lower glomerular IC deposition is associated with a reduced inflammatory response and impaired organ damage. The reduced frequency of GN in SLE patients who have IgM anti-dsDNA antibodies may therefore reflect a disease modifying effect of this class of autoantibodies that has potential therapeutic implications. Our findings should encourage the development of new therapeutic modalities using IgM anti-dsDNA antibodies in humans with SLE. PMID- 16255056 TI - Increased expression of the novel proinflammatory cytokine high mobility group box chromosomal protein 1 in skin lesions of patients with lupus erythematosus. AB - OBJECTIVE: To investigate the role of the novel cytokine high mobility group box chromosomal protein 1 (HMGB-1) in the pathogenesis of cutaneous lupus erythematosus (CLE). METHODS: Punch biopsy specimens of lesional and unaffected skin from 10 patients with CLE and 3 healthy control subjects were investigated. Immunohistochemical staining for HMGB-1, tumor necrosis factor alpha (TNFalpha), and interleukin-1beta (IL-1beta) was performed on consecutive sections. Analysis of single-nucleotide polymorphisms of -308 TNF was performed on DNA extracted from peripheral blood mononuclear cells. RESULTS: An altered expression of HMGB-1 was observed both in the epidermis and in the dermal infiltrates of lesional skin. Expression of HMGB-1 in the epidermis and dermis was increased (P < 0.01 and P < 0.001, respectively, versus unaffected skin), and translocation to the cytoplasm as well as the extracellular presence of secreted HMGB-1 were found. Increased levels of TNFalpha and IL-1beta were also observed in the dermal infiltrates of lesional skin (P < 0.01 and P < 0.05, respectively, versus unaffected skin). The carrier frequency of the -308A TNF polymorphism was 80% in patients with subacute CLE, but this was not related to higher expression of TNFalpha in biopsy specimens from the CLE group. CONCLUSION: The high amount of extracellular HMGB-1 observed in skin lesions indicates that HMGB-1 is involved in the inflammatory process of CLE. TNFalpha and IL-1beta may form a proinflammatory loop with HMGB-1, since they can induce the release of each other. The extracellular HMGB-1 observed by immunostaining of the epidermis indicates that keratinocytes may be an as yet unrecognized source of secreted HMGB-1, underscoring the role of the target organ in the rheumatoid autoimmune inflammatory process. PMID- 16255057 TI - Influence of mechanical stress and surface interaction on the aggregation of Aspergillus niger conidia. AB - Productivity of fungal cultures is closely linked with their morphologic development. Morphogenesis of coagulating filamentous fungi, like Aspergillus niger, starts with aggregation of conidia, also denominated as spores. Several parameters are presumed to control this event, but little is known about their mode of action. Rational process optimization requires models that mirror the underlying reaction mechanisms. An approach in this regard is suggested and supported by experimental data. Aggregation kinetics was examined for the first 15 h of cultivation under different cultivation conditions. Mechanical stress was considered as well as pH-dependent surface interaction. Deliberations were based on a two-step aggregation mechanism. The first aggregation step is only affected by the pH-value, not by the fluid dynamic conditions in the bioreactor. The second aggregation step, in contrast, depends on the pH-value as well as on agitation and aeration induced power input. For the given experimental set-up, agitation had much more influence than aeration. In addition, hyphal growth rate was determined to be the driving force for the second aggregation step. PMID- 16255058 TI - Enhancement of protein secretion in Pichia pastoris by overexpression of protein disulfide isomerase. AB - A potential vaccine candidate, Necator americanus secretory protein (Na-ASP1), against hookworm infections, has been expressed in Pichia pastoris. Na-ASP1, a 45 kDa protein containing 20 cysteines, was directed outside the cell by fusing the protein to the preprosequence of the alpha-mating factor of Saccharomyces cerevisiae. Most of the protein produced by single copy clones was secreted outside the cell. However, increasing gene copy number of Na-ASP1 protein in P. pastoris saturated secretory capacity and therefore, decreased the amount of secreted protein in clones harboring multiple copies of Na-ASP1 gene. Overexpression of the endoplasmic reticulum (ER) resident, homologous chaperone protein, protein disulfide isomerase (PDI) was able to increase the secretion of (Na-ASP1) protein in high copy clones. The effect of PDI levels on secretion of Na-ASP1 protein was examined in clones with varying copy number of PDI gene. Increase in secreted Na-ASP1 secretion is correlated well with the PDI copy number. Increasing levels of PDI also increased overall Na-ASP1 protein production in all the clones. Nevertheless, there was still accumulation of intracellular Na-ASP1 protein in P. pastoris clones over-expressing Na-ASP1 and PDI proteins. PMID- 16255059 TI - Linkage at 5q14.3-15 in multiplex systemic lupus erythematosus pedigrees stratified by autoimmune thyroid disease. AB - OBJECTIVE: To identify genetic effects potentially shared between systemic lupus erythematosus (SLE) and autoimmune thyroiditis (AITD). METHODS: Families from the Lupus Multiplex Registry and Repository were studied in which there was at least 1 member who had both SLE and AITD (Graves' disease or Hashimoto thyroiditis). Genome scan genotyping findings in these pedigrees were evaluated for evidence of genetic linkage, by the maximum-likelihood parametric method. Nineteen pedigrees were used in the initial genome scan. Subsequently, an independent sample of 16 pedigrees was used to replicate findings. RESULTS: Studies of the first set of 19 pedigrees yielded a 2-point parametric logarithm of odds (LOD) of 4.97, which was independently confirmed in the replication sample of 16 pedigrees (LOD 2.89). For all 35 pedigrees together, the 2-point LOD was 7.86, under a dominant model used for screening with 90% penetrance and a disease allele frequency of 10%. The multipoint locus homogeneity LOD in the 35 pedigrees was 6.90 (alpha = 1.0) at 5q14.3-15 between D5S1725 and D5S1453, a 12-cM interval, with the peak at D5S1462 at 96.64 cM (nonparametric linkage P = 0.00002). Fine mapping further confirmed the genetic linkage effect and narrowed the region likely to contain the gene to approximately 5 Mb. CONCLUSION: These results suggest that stratifying SLE pedigrees by the presence of other autoimmune disorders may facilitate the discovery of genes related to SLE and that 5q14.3-15 harbors a susceptibility gene shared by SLE and AITD. PMID- 16255060 TI - Avilamycin is poorly absorbed, is extensively metabolized by swine and is not persistent in the environment. PMID- 16255061 TI - Ion/molecule reactions in a miniature RIT mass spectrometer. AB - Ion/molecule reactions were explored in a newly developed miniature mass spectrometer fitted with a rectilinear ion trap (RIT) mass analyzer. The tandem mass spectrometry performance of this instrument is demonstrated using collision induced dissociation (CID) and ion/molecule reactions. The latter includes Eberlin transacetalization reactions and electrophilic additions. Selective detection of the chemical warfare simulant dimethyl methyl phosphonate (DMMP) was achieved through selective Eberlin reactions of its characteristic phosphonium fragment ion CH3OP(+)(O)CH3 (m/z 93), with 1,4-dioxane or 1,3-dioxolane. Efficient adduct formation as a result of electrophilic attack by the phosphonium ion on various nucleophilic reagents, including 1,1,3,3-tetramethyl urea, methanesulfonic acid methyl ester, dimethyl sulfoxide and methyl salicylate, was also observed using the RIT device. The product ions of these reactions were analyzed using CID and the characteristic fragmentation patterns of the ionic addition products were recorded using multiple-stage experiments in the miniature RIT instrument. This study clearly demonstrates that a small, home-built, miniature RIT mass spectrometer can be used to perform analytically useful ion/molecule reactions and also that instruments like this have the potential to provide a portable platform for in situ detection of organophosphorus esters and related compounds with high specificity using tandem mass spectrometry. PMID- 16255062 TI - Screening of organic nitrate explosives: selective ion/molecule reactions for the diagnostic ion NO2+. PMID- 16255063 TI - Simultaneous determination of metformin and gliclazide in human plasma by liquid chromatography-tandem mass spectrometry: application to a bioequivalence study of two formulations in healthy volunteers. AB - A rapid and sensitive liquid chromatography/tandem mass spectrometry (LC/MS/MS) method was developed and validated to simultaneously determine gliclazide and metformin in human plasma using huperzine A as the internal standard (IS). After acetonitrile-induced protein precipitation of the plasma samples, gliclazide, metformin and the IS were subjected to LC/MS/MS analysis using electro-spray ionization (ESI). Chromatographic separation was performed on a Hypersil BDS C18 column (50 mm x 2.1 mm, i.d., 3 microm). The method had a chromatographic running time of 2.0 min and linear calibration curves over the concentration ranges of 10 10,000 ng ml(-1) for gliclazide and 7.8-4678.9 ng ml(-1) for metformin. The recoveries of the method were found to be 71-104%. The lower limits of quantification (LOQ) of the method were 10.0 and 7.8 ng ml(-1) for gliclazide and metformin, respectively. The intra- and interday precision was less than 15% for all quality control samples at concentrations of 100, 500, and 2000 ng ml(-1). The validated LC/MS/MS method has been used to study bioequivalence in healthy volunteers. These results indicate that the method was efficient with a very short running time (2.0 min) for metformin and gliclazide compared to the methods reported in the literature. The presented method had acceptable accuracy, precision and sensitivity and was used in clinical bioequivalence study. PMID- 16255064 TI - New warning on antipsychotic drugs used to treat older people. PMID- 16255065 TI - Final rule on albuterol inhalers. PMID- 16255066 TI - New treatment for chronic hepatitis B. PMID- 16255067 TI - Personalized medicine. PMID- 16255068 TI - HHS buys children's version of radiation emergency drug. PMID- 16255070 TI - New oral rinse helps treat gingivitis. PMID- 16255069 TI - Counterfeit drug warning. PMID- 16255071 TI - Empowering Family Doctors and Patients in Nutrition Communication. Proceedings of an international workshop, December 13-15, 2004, Heelsum, The Netherlands. PMID- 16255072 TI - Effect of dietary polyunsaturated fatty acids on age-related changes in cardiac mitochondrial membranes. AB - Remodeling of myocardial cell membranes is a major feature of advanced age. Mitochondrial function, crucial to sustaining energy production and management of myocardial metabolism, is impacted by age-dependent remodeling and ultimately exhibits a diminished threshold for excess Ca2+ buffering during events that stimulate increased myocardial Ca2+, such as augmented cardiac work, oxidative stress or post-ischemic reflow. Relative Ca2+, intolerance, augmented superoxide formation and reduced efficiency in the management of reactive oxygen species, are important mitochondrial factors (of many) that are apparent in senescence and predispose the myocardium to be more vulnerable to ischemic injury. In addition to cell death, surviving myocytes increase in size and exhibit altered gene expression of key effector proteins, including those that sustain Ca2+ homeostasis. Age-associated mitochondrial membrane changes include increases in membrane rigidity, cholesterol, phosphatidylcholine, omega-6 polyunsaturated fatty acids (PUFA), 4-hydroxy-2-nonenal, and decreases in omega-3 PUFA and cardiolipin. These effects have been shown in animal studies to be exaggerated by diet rich in long chain omega-6 PUFA (i.e. arachidonic acid), and have profound consequences on the efficacy of membrane proteins involved with ion homeostasis, signal transduction, redox reactions and oxidative phosphorylation. However, some of the age-related detrimental adaptations may be beneficially modified by dietary strategy. Diet rich in omega-3 PUFA reverses the age-associated membrane omega-3:omega-6 PUFA imbalance, and dysfunctional Ca2+ metabolism, facilitating increased efficiency of mitochondrial energy production and improved tolerance of ischemia and reperfusion. PMID- 16255073 TI - [Organization and role of the German public health service]. AB - The article describes the organization and role of the public health service in Germany. Health is not a federal but a state issue; sta-te health regulations are subject to state legislation. The role of the public health service may therefore differ among the federal sta-tes of Germany. In recent years there ha-ve been considerable changes in the main features of the role of the public health ser vice. Prevention and health reporting have become top issues, as well as hygiene, control of epidemics, medical certificates, and physical and dental examinations for schoolchildren. The public health service has developed into an up-to-date and customer-focused organization. Using modern management methods and employing well-trained doctors and other highly skilled health professionals, the public health service will full-fill its role in working for the benefit of the citizens of Germany providing the necessary resources are available. The generational change in the public health service will be marked in the coming years, especially in the east of Germany. Many physicians will reach retirement age and need replacing. Physicians with postgraduate professional education in public health are rare; training in this area of expertise must be fostered. PMID- 16255075 TI - [Prevention of diseases and health monitoring by the local public health services. Tasks and prospects]. AB - For many years prevention of disease and health promotion have been central activities of the public health services in Germany. But especially within these areas of activity there have been difficulties in understanding "old"versus"new"in relation to public health. Previously it was not possible to generate common standards for the public health service for prevention, health promotion and health monitoring due to the regional diversity of legislation within the single provinces. However, these activities could demonstrate the strength of public health services in these fields. The forthcoming law on prevention provides a framework for strengthening the public health services through defining qualitative standards for eligible preventive measures and at the same time making clear the competences of public health services that should be available. Thus public health services could become one of the central players at the local level in the strengthening of prevention and health promotion in the future. The quality of planning and outcome of eligible preventive measures will strongly depend on the local health monitoring system. If the forthcoming law does not make use of the competence of public health services in identifying healthy and socially disadvantaged settings, a white-collar orientation of preventive and health promotion activities may be expected. PMID- 16255074 TI - [Responsibilities of the Public Health Service in psychiatric care]. AB - Almost unnoticed by the general public, but also to some extent by public health professionals, psychiatric care in Germany has recently undergone considerable change. This development could be considered as more revolutionary than changes seen in somatic medicine. These changes can also be found in progress in "psychiatry as a science"in areas as diverse as prevention, diagnosis and treatment of those with mental illness, and their rehabilitation, recovery and after-care. Public health professionals have long witnessed these changes which have fluctuated in intention and intensity. This paper reviews the development of present-day responsibilities of the Public Health Service in psychiatric care and underlines the need for reform presenting an implementation plan and an outline of possible structural changes. PMID- 16255076 TI - [Do both the public health service and public health have a future despite developing in different directions?]. AB - In the United Kingdom and the United States the public health services have always been considered as being an integral part of public health. The mission, tasks and objectives are the same. In Germany pub-lic health research and public health initiatives on behalf of the universities started in the 1990s. Before that terms and disciplines such as "hygiene" and "social medicine" prevailed. The public health service -- related to these disciplines -- had also been established previously. At the start of the debate on public health issues, however, the pub-lic health service did not play an important role. It was criticized for its conventional organization and its lack of population-based strategies. This article describes the conceptual development of both public health and the public health service over the last 15 ye-ars and the importance of public health politics in achieving a joint strategy aimed at im-proving the health of the population including different population groups. PMID- 16255077 TI - Abstracts of the American Association for Cancer Education 39th Annual Meeting. September 15-17, 2005, Cincinnati, Ohio, USA. PMID- 16255078 TI - [Contribution of the Public Health Service to cost of illness studies]. AB - This paper reflects on the different approach-es of top-down and bottom-up analysis when calculating sickness costs. Following an analysis of the standard procedures to be followed in tuberculosis contact tracing performed by public health bureaux in Hamburg, a top-down estimate is presented de-spite some accounting limitations. The val-ue of epidemiologic data on infectious dis-eases held by the Public Health Service for bottom-up-modelling is shown by a cost-of illness study on hepatitis A in Germany.However, broader efforts may be required by local service providers to justify their existence from an economic perspective. This could also contribute to improving cooperation between public health authorities and schools of public health in this rapidly developing scientific field. PMID- 16255079 TI - Hospital care of patients with ST-elevation myocardial infarction in four different European regions. Data from four regional myocardial infarction registries in Berlin, Dijon, Florence, and Tartu. AB - National surveys as well as European comparative studies suggest that differences in treatment of patients with ST-elevation myocardial infarction (STEMI) exist. The extent to which these variations influence the outcome of hospital care delivered to STE-MI patients in everyday routine is mostly unknown. In this study data representative of hospital care received by STEMI patients in four European regions (Berlin, Dijon, Florence and Tartu) were compared. The four registries are population based. The percentage of women and the mean age of the patients differed among the registries. Risk factors such as hypertension and hypercholesterolaemia also differed among the different regions, whereas a history of diabetes mellitus was similar among the registries. The percentage of patients receiving reperfusion therapy ranged from 47 to 81%. An appreciable difference also resulted after breaking down reperfusion therapy into thrombolysis and primary percutaneous coronary intervention (PCI). Hospital mortality as an outcome measure was very similar among the regions. After adjustment for age, the comparative magnitude of hospital mortality proportion was also very similar among three registries. Only the patients from Florence demonstrated a comparatively lower death rate, with a ratio of 0.81. In summary, there are important differences among baseline characteristics and hospital care of STE-Ml patients in the four study regions. Nevertheless, it was interesting to ascertain that the outcome measured in hospital mortality was very similar among the four registries compared. PMID- 16255081 TI - Preparation and characterization of melittin-loaded poly (DL-lactic acid) or poly (DL-lactic-co-glycolic acid) microspheres made by the double emulsion method. AB - The water soluble peptide, melittin, isolated from bee venom and composed of twenty-six amino acids, was encapsulated in poly (DL-lactic acid, PLA) and poly (DL-lactic-co-glycolic acid, PLGA) microspheres prepared by a multiple emulsion [(W1/O)W2] solvent evaporation method. The aim of this work was to develop a controlled release injection that would deliver the melittin over a period of about one month. The influence of various preparation parameters, such as the type of polymer, its concentration, stabilizer PVA concentration, volume of internal water phase and level of drug loading on the characteristics of the microspheres and drug release was investigated. It was found that the microspheres of about 5 microm in size can be produced in high encapsulation (up to 90%), and the melittin content in the microspheres was up to 10% (w/w). The drug release profiles in vitro exhibited a significant burst release, followed by a lag phase of little or no release and then a phase of constant melittin release. The type of polymer used was a critical factor in controlling the release of melittin from the microspheres. In this study, the rate of peptide release from the microspheres correlated well with the rate of polymer degradation. Moreover, melittin was released completely during the study period of 30 days, which agreed well with the polymer degradation rate. PMID- 16255082 TI - Abstracts of the 8th Congress of the European Association of Clinical Anatomy (EACA), Palermo, Italy, June 30-July 3, 2005. PMID- 16255083 TI - Abstracts of the XIII World Congress on Psychiatric Genetics 2005. PMID- 16255080 TI - A haplotype map of the human genome. AB - Inherited genetic variation has a critical but as yet largely uncharacterized role in human disease. Here we report a public database of common variation in the human genome: more than one million single nucleotide polymorphisms (SNPs) for which accurate and complete genotypes have been obtained in 269 DNA samples from four populations, including ten 500-kilobase regions in which essentially all information about common DNA variation has been extracted. These data document the generality of recombination hotspots, a block-like structure of linkage disequilibrium and low haplotype diversity, leading to substantial correlations of SNPs with many of their neighbours. We show how the HapMap resource can guide the design and analysis of genetic association studies, shed light on structural variation and recombination, and identify loci that may have been subject to natural selection during human evolution. PMID- 16255085 TI - John H. C. Ho. PMID- 16255084 TI - Creating a crosswalk to estimate AIDS Clinical Trials Group quality of life scores in a nationally representative sample of persons in care for HIV in the United States. AB - BACKGROUND: Optimizing health-related quality of life (HRQOL) is an increasingly important goal in the treatment of HIV/AIDS. Interpretation of HRQOL scores in clinical trials is enhanced by comparative data. PURPOSE: To estimate AIDS Clinical Trials Group (ACTG) QOL 601-602 questionnaire scale scores for a nationally representative sample of persons in care for HIV. METHOD: The study cohort was from the HIV Cost and Services Utilization Study (HCSUS), a multistage national probability sample. We derived HCSUS HRQOL scale scores from the items shared between the ACTG QOL 601-602 and HCSUS HRQOL questionnaires using regression equations. Cronbach's alpha coefficient was used to estimate the reliability of the multi-item scales in the ACTG QOL 601-602 and HCSUS HRQOL instruments. Correlation Coefficients and R2s of regression models were calculated to determine the concordance of the models. Multiple regression was used to determine if patient characteristics accounted for differences (residuals) between scores observed from the full HCSUS HRQOL instruments and scores predicted using the subset of shared items in ACTG QOL 601-2. RESULTS: Internal consistency reliability estimates were acceptable (>0.70) for all scales in the ACTG QOL 601-602 and HCSUS HRQOL instruments. Correlations between corresponding ACTG QOL 601-602 and HCSUS HRQOL scale scores were high (>0.9). The R2s for predicting HCSUS HRQOL scores from the ACTG QOL 601-602 scales were also high (>0.8). For physical functioning, emotional well-being, and general health perceptions, the predictors of differences (residuals) in observed and predicted HCSUS HRQOL scores were gender and CDC stage of HIV infection (P < .05). CONCLUSION: This study provides normative data from the US HIV/AIDS population for comparison to the ACTG QOL 601-602 questionnaire. Accuracy of estimation is enhanced if done separately by gender and HIV disease stage. PMID- 16255086 TI - Interradicular radiolucency of mandible. PMID- 16255087 TI - Katrina's digital dividend. Feds want damaged hospitals rebuilt with EMRs. PMID- 16255088 TI - Healthcare shopping. Community Health, HMA to buy, upgrade hospitals. PMID- 16255089 TI - Comprehensive modeling of the temperature-related laser performances of the amplifiers of the LUCIA laser. AB - We present numerical simulations of the temperature-related laser performance of the amplifiers for the Lasers Ultra-Courts et Intenses et Applications (LUCIA) laser, a 100 J, 10 Hz, 10 ns diode-pumping solid-state-laser facility, which uses Yb3+:YAG as the gain medium. The simulations include energy storage and extraction efficiency, cooling of the gain medium, and wavefront distortion. The results show that, with a pumping intensity of 20 kW/cm2 at 10 Hz and a doping concentration of 10 at. % at a thickness of Yb3+:YAG of 1.6 mm, the output laser fluence and optical-to-optical efficiency are expected to be 10 J/cm2 and 25.8%, respectively, at a heat exchange coefficient of 3000 W/m2/K of water. Also, the matching thickness of undoped YAG is optimized to prevent bending deformation of the gain medium, which could be approximately 5 mm. PMID- 16255090 TI - Findings of scientific misconduct. PMID- 16255091 TI - Statins: adjunctive pharmacotherapy for percutaneous coronary intervention? PMID- 16255092 TI - Diuretic therapy, natriuretic peptides, and heart failure. PMID- 16255093 TI - Current awareness on yeast. PMID- 16255094 TI - Coffee vs. tea. Benefits and risks. PMID- 16255095 TI - Job cut will pile on the pressure. PMID- 16255098 TI - A century of care. PMID- 16255099 TI - Tackling taboos on prison abuse. Interview by Karen Kelly. PMID- 16255100 TI - The nurse's role in promoting breast awareness to women. AB - Breast awareness is now a recognised part of health promotion and has replaced previous guidance on breast self-examination for women. Since it was introduced in the early 1990s, along with the breast awareness five-point code, it has become a key element in the strategy to detect breast cancer at an early stage. Despite this, confusion about breast awareness persists, with women still wanting to know how to examine their breasts properly. PMID- 16255101 TI - Verocytotoxin-producing Escherichia coli. PMID- 16255102 TI - Blood cells. Part two--Red blood cells. PMID- 16255103 TI - Taking resuscitation decisions in the nursing home setting. AB - The way decisions about resuscitation are made in A&E departments is not the same as in nursing homes for older people. This article discusses prompt appropriate decision-making regarding resuscitation in the nursing home setting. PMID- 16255104 TI - Methylprednisolone. PMID- 16255105 TI - Analysing and improving a rapid-access chest pain clinic. AB - This article describes how involving patients and analysing procedures in a rapid access chest pain clinic improved the service. Waiting times were reduced, targets were met, processes were improved and a new cardiac rehabilitation service was offered to patients with newly diagnosed angina. PMID- 16255106 TI - The utility of CPD for older adult mental health nurses. AB - AIM: To investigate how mental health nurses working with older adults perceive the benefits and realities of developing the outcomes of current continuing professional development training into actual clinical practice. METHOD: A structured questionnaire was used with a convenience sample of nursing staff. Qualitative analysis was performed using a grounded theory approach in order to identify emergent themes, concepts and categories of data. Four randomly selected nurses were subjected to a voluntary semistructured interview using the questionnaire as a basis for information gathering. RESULTS: The main reason for attending courses was developing skills. Of those attending courses, 42 per cent of qualified and 35 per cent of unqualified staff had a personal development plan (PDP) or individual performance review (IPR). Significantly, all unqualified staff who had not been on a course had no PDP or IPR. Learning was described as applicable to practice by 85 per cent of unqualified and 70 per cent of qualified staff. However, 28 per cent of unqualified staff and 20 per cent of qualified staff felt their practice had not changed as a result of their learning. CONCLUSION: CPD can be a positive experience, providing nurses with the opportunity to direct their professional development. PMID- 16255107 TI - How to be successful in senior job interviews. PMID- 16255108 TI - Where patient trust is essential. PMID- 16255109 TI - Emotional and practical support to get through day-to-day life. PMID- 16255110 TI - Health screening tests: a note of caution. PMID- 16255111 TI - Uterine rupture after a previous Caesarean section. PMID- 16255112 TI - Alcohol and heart disease--what do you prescribe? PMID- 16255113 TI - A survey of the causes of sudden cardiac death in the under 35-year-age group. AB - CRY (Cardiac Risk in the Young) is a registered Irish charity established by parents who are bereaved as a result of sudden cardiac death. The aim of this study is to establish the incidence and causes of sudden cardiac death in Dublin city in the 10-year period from 1st January 1993 to 31st December 2002. All sudden cardiac deaths in the under 35-year age group which were reported to the city coroner in the study period were examined. Details regarding age, sex, previous symptoms, investigations, circumstances of death and main pathological finding were recorded in each case. A total of 72 cases of sudden cardiac death in the under-35 year age group were reported. 52 were men. The median age was 26.5 years (range 12-34 years). The cause of death in 20 cases was reported as atherosclerotic Coronary Artery Disease. The second commonest cause of death (24% cases) was Hypertrophic Cardiomyopathy. Hypertrophic Cardiomyopathy was the commonest cause of death under the age of 25 years. Overall atherosclerotic coronary artery disease was the commonest cause of death in this group. The importance of Hypertrophic Cardiomyopathy is highlighted by the fact it was the commonest cause of death in the under 25-year age group. Screening those at high risk of sudden cardiac death especially the relatives of those affected by Hypertrophic Cardiomyopathy need to be discussed and implemented. PMID- 16255114 TI - Multi organ transplantation: a review of the Irish experience. AB - As the criteria for organ transplantation have broadened, multi organ transplantation has become more common. Calcineurin inhibitor induced end stage renal failure is a common indication for kidney transplantation. We present our experience of 12 kidney transplants, in 11 patients, over a period of 9 years all in the setting of multi-organ transplantation. Kaplan-Meier survival estimates were applied. Estimated patient survival at 1, 3 and 5 years post transplant was 100%, 83% and 66% respectively. Estimated graft survival at 1, 3 and 5 years was 90%, 79% and 63% respectively. The estimated median renal graft survival was 6.2 years. Calcineurin inhibitor toxicity was the most common cause of end stage renal failure in this series of patients. There is a very fine balance between ideal immunosuppression for kidney and other organ transplants. Patients who avoid morbidity such as sepsis or graft rejection enjoy good graft function at 5 years post transplant. PMID- 16255115 TI - Delays in treatment in the cancer services: impact on cancer stage and survival. AB - This study describes waiting times for cancer treatment in Ireland, tests for differences between health board areas and investigates if delay affected cancer stage or prognosis. We measured the times between referral, outpatient attendance and treatment for all patients diagnosed with five common cancers in Ireland in 1999. There was a four-fold spread of waiting time between the 25th and 75th centile. The time from referral to treatment was shortest for breast cancer (median 4 weeks), twice this for colorectal cancer, lung cancer and lymphoma, and 15 weeks for prostate cancer. Delays varied significantly, but not consistently, between health boards. Patients with longer waiting times generally had less advanced disease and better survival, suggesting that typical delays are not of clinical significance, but that patients with advanced disease are probably being "fast-tracked" by GPs and hospitals. PMID- 16255116 TI - Mucinous neoplasm of the appendix with elevated carcinoembryonic antigen. PMID- 16255118 TI - Acute gastric dilatation in a child with marked kyphoscoliosis and cerebral palsy -a case report. PMID- 16255117 TI - Rapid cardioversion of new onset atrial fibrillation with ibutilide in the emergency department. AB - People with atrial fibrillation commonly present to the emergency department. Ibutilide is an anti arrhythmic indicated for the cardioversion of atrial fibrillation and flutter where the onset is less than 48 hours. PMID- 16255119 TI - Axillary lymphadenopathy secondary to cat-scratch disease. PMID- 16255120 TI - Primary Hepatic Lymphoma. PMID- 16255121 TI - Thoracic outlet syndrome secondary to first rib anomaly: the value of multi-slice CT in diagnosis and surgical planning. PMID- 16255122 TI - Two cases of meralgia paraesthetica following open appendicetomy. PMID- 16255123 TI - Influences of emotion on context memory while viewing film clips. AB - Participants listened to words while viewing film clips (audio off). Film clips were classified as neutral, positively valenced, negatively valenced, and arousing. Memory was assessed in three ways: recall of film content, recall of words, and context recognition. In the context recognition test, participants were presented a word and determined which film clip was showing when the word was originally presented. In two experiments, context memory performance was disrupted when words were presented during negatively valenced film clips, whereas it was enhanced when words were presented during arousing film clips. Free recall of words presented during the negatively valenced films was also disrupted. These findings suggest multiple influences of emotion on memory performance. PMID- 16255124 TI - Inattentional blindness for a noxious multimodal stimulus. AB - Previous research has shown that people can miss salient stimuli outside the focus of their attention. This phenomenon, called inattentional blindness, typically is observed when people are given a task requiring them to focus their attention on one aspect of a complex visual scene. While participants are doing this task, an unexpected stimulus appears, and participants' awareness of it is tested shortly thereafter. In the present experiments, noxious bimodal stimuli were used as a test case to measure the strength of inattentional blindness. We tested whether participants would notice a person enter a scene and scratch her fingernails down a chalkboard (thus making a sound called a "gride"). A large proportion of participants failed to detect this event even when the noxious audio associated with it was strengthened and isolated in time from surrounding noises. PMID- 16255125 TI - Consistent contrast and correlation in free sorting. AB - Two experiments investigated free sorting, a type of unsupervised learning, with multiattribute drawings of alien animals. In previous research on concept formation, with simpler stimulus structure than ours, participants were insensitive to correlational structure in the stimuli, producing primarily "ID sorts," based on the values ofjust 1 dimension or attribute. Our experiments showed that participants used many strategies in categorizing but preferred to generate groupings that reflected the correlations in input when this did not violate consistent contrast. The second experiment used hierarchically structured stimuli to show that participants' sort strategies favor consistency within a set of contrasting categories, distinct from any preference for 1D sorting. Finally, both experiments show that correlational sorts are much more likely when the correlation-based sort contrasts consistently. Our data show complexity at work in free sort tasks: People are sensitive to multiple and sometimes conflicting biases for consistency and correlational structure in the category systems they create. PMID- 16255126 TI - The influence of stimulus array on training of a speeded response. AB - We conducted two experiments to study the training of speeded responses to stimuli in a clock face array. Participants were trained on one clock face and later tested on either the same or a different clock face. The clock faces varied along 3 dimensions of change (left-right flip, up-down flip, and 180 degrees rotation). Skill acquisition was reflected in decreases in overall, initiation, and movement response times in the training and test phases except that movement time reached an asymptote during training and did not decrease further during test. Test performance was best when training and test used the same clock face and worst when the training to test change involved an up-down flip. The transition to different clock faces was consistent with an interpretation based on the violation of expectations about target positions acquired during training. Consistent trial-by-trial feedback led to better initiation time but not movement time during training than periodic feedback, but this effect did not persist into the test phase. PMID- 16255127 TI - Practice effects, workload, and reaction time in deception. AB - Cognitive theorists argue that deception may involve attentional capture, working memory load, or perceived incongruity with memory, whereas psychophysiologists argue for stimulus salience, arousal, and emotion. This article presents a comprehensive model of deception and assesses the effect of practice on deceptive responding. A three-session longitudinal study examined the effect of practice on reaction time (RT) in relation to deception and response congruity. Participants evaluated self-referent sentences and responded truthfully or deceptively. Findings indicate that deceptive responding generates longer RTs than does truthful responding and that this relationship remains constant with practice. We use these findings to support a cognitive model of deception. PMID- 16255128 TI - Evidential reasoning in the prisoner's dilemma. AB - Classic game theory considers defection to be the rational choice in the Prisoner's Dilemma. Although defection maximizes a player's gain regardless of the opponent's choice, many players cooperate. We suggest that cooperation can be explained in part by expectations of reciprocal behavior, that is, by the belief that players facing the same situation probably will come to the same decision. In Experiment 1, expectations of reciprocity were experimentally manipulated. As predicted, cooperation increased monotonically with these expectations. In Experiment 2, experimentally manipulated expectations of opponent cooperation were not associated with higher rates of cooperation. These findings are interpreted in light of a general model of payoff maximization, and implications for other models (e.g., social value orientation) are discussed. PMID- 16255129 TI - The Howard Crosby Warren Medal: psychology's first award. AB - This article explores the development of the first major award given in American psychology, the Howard Crosby Warren Medal. Administered by the Society of Experimental Psychologists, the award was first given in 1936. The first recipients of the Howard Crosby Warren Medal were Ernest G. Wever and Charles W. Bray of Princeton University. The Howard Crosby Warren Medal remains among the most prestigious awards in American psychology. PMID- 16255130 TI - Clinical trials referral resource. Current clinical trials of erlotinib. PMID- 16255131 TI - Interim data suggest major response with darbepoetin in anemic MDS patients. PMID- 16255132 TI - Metastatic colorectal cancer: is there one standard approach? AB - Despite enormous advances in the treatment of colorectal cancer, there is no single standard treatment approach for all patients. However, there are general principles of management that can be used to guide therapy. The clinician who fails to individualize therapy for colorectal cancer is likely not taking full advantage of all therapeutic options available. Reviewing key clinical evidence that can help inform decision-making, this article addresses important questions in colorectal cancer management, including: Should bevacizumab (Avastin) be a component of most patients' first-line treatment? Is there a role for continuing bevacizumab in subsequent regimens? Is there a role for cetuximab (Erbitux) in standard first-line chemotherapy? Are there practices in colorectal cancer that have become widely accepted without direct supportive data? PMID- 16255133 TI - Mohs micrographic surgery: established uses and emerging trends. AB - Mohs micrographic surgery is a surgical technique that seeks to ensure the clearance of cutaneous tumors while maximizing normal tissue conservation. This is accomplished through the sequential removal of thin layers of tissue in which the entire peripheral and deep margins are examined for residual tumor. This approach appears to be superior to conventional surgical excision in the treatment of basal cell carcinoma (BCC) and squamous cell carcinoma (SCC), the two most common cancers of the skin. Its efficacy in treating BCC and SCC has led clinicians to explore the role of Mohs micrographic surgery in the management of less common cutaneous neoplasms, such as melanoma, Merkel cell carcinoma, dermatofibrosarcoma protuberans, extramammary Paget's disease, and microcystic adnexal carcinoma. PMID- 16255134 TI - Biphasic tumors of the female genital tract. PMID- 16255135 TI - Cancer management in patients with end-stage renal disease. AB - Significant improvements in the management of patients with end-stage renal disease (ESRD) who are on chronic renal replacement therapy (CRRT), has led to an increase prevalence of this population among older Americans. Since cancer is also common in the elderly, oncologists are likely to be faced with patients who suffer from both cancer and ESRD. There is a paucity of information regarding issues surrounding the optimal management of such patients, especially those needing chemotherapy. This review surveys the relevant problems oncologists may encounter in such patients and summarizes the available literature on chemotherapeutic management of common cancers. The reader is strongly urged to consult the original references for details of chemotherapy administration prior to use in an individual patient. PMID- 16255136 TI - Neoadjuvant therapy for gastric cancer. AB - Gastric cancer is a global health issue. Most cases are diagnosed at an advanced stage with poor prognosis. Current therapies have a modest impact on survival. Surgery remains the only potentially curative treatment, but is associated with a high rate of locoregional recurrence and distant metastases. Total gastrectomy for proximal cancers is complicated by postoperative morbidity and quality-of life impairment. Combined-modality therapy may improve outcomes in this disease. Adjuvant therapy for gastric cancer has now become the standard in the Western world. However, adjuvant therapy improves survival by only a few months and is associated with high morbidity. Neoadjuvant therapy is commonly used for esophageal and gastroesophageal junction cancers, but is still regarded as investigational in gastric cancer. Several small phase II studies indicate the feasibility of neoadjuvant strategies. The incorporation of novel, targeted agents into neoadjuvant programs and an assessment of biologic changes within the tumor may refine therapy. This article provides a concise review of the literature on neoadjuvant therapy for gastric cancer and suggests avenues for further investigation. PMID- 16255137 TI - Effect of carbon source on pyrimidine formation in Pseudomonas fluorescens ATCC 13525. AB - The effect of carbon source on the regulation of the de novo pyrimidine biosynthetic enzymes in the type strain of Pseudomonas fluorescens was investigated. The de novo pyrimidine biosynthetic enzyme activities were measured in P. fluorescens ATCC 13525 cells and in cells from an auxotroph deficient for orotate phosphoribosyltransferase activity where glucose or succinate served as the carbon source. Pyrimidine supplementation to the culture medium affected the biosynthetic enzyme activities in ATCC 13525 cells. Pyrimidine Limitation of the mutant strain indicated transcriptional regulation of the biosynthetic pathway by pyrimidines that was influenced by carbon source. Transcriptional regulation of pyrimidine synthesis in P. fluorescens appeared to be more highly regulated than was observed for the taxonomically related species Pseudomonas aeruginosa or Pseudomonas putida. PMID- 16255138 TI - Biological control of bacterial spot of tomato caused by Xanthomonas campestris pv. vesicatoria by Rahnella aquatilis. AB - Xanthomonas campestris pv. vesicatoria strain 2 was isolated from infected tomato seedlings grown in open field in Egypt. This strain produced irregular yellow necrotic areas on tomato leaves and spotting of the stem. In an attempt to control this disease biologically, four experiments were conducted and tomato seedlings were pretreated, before the pathogen, with either of two antagonistic strains of Rahnella aquatilis through leaves, roots, soil or seeds. In all experiments, seedlings pretreated with R. aquatilis showed reduced susceptibility toward X. c. pv. vesicatoria. They also contained reduced protein concentration and showed reduced number of protein bands in SDS-PAGE analysis as well as increased fresh and dry weight relative to control seedlings inoculated with the pathogen only. This indicates that R. aquatilis reduced the deleterious effect and the stress exerted by X. c. pv. vesicatoria on tomato seedlings. Foliar application of R. aquatilis was the most effective method in disease reduction which could be attributed to the direct effect of the antagonistic bacteria on the pathogen. The highest amounts of fresh and dry weight were obtained from seed treatment, which might suggest that bacterial seed inoculation provides earlier protection than could be achieved with foliar, soil or root treatment. PMID- 16255139 TI - Contamination of malt barley and wheat by Fusarium graminearum and Fusarium culmorum from the crop years 2001-2003 in eastern Croatia. AB - This study investigated infection levels with Fusarium graminearum and Fusarium culmorum in malt barley and wheat in eastern Croatia. The contamination was surveyed over three consecutive crop years (2001-2003) on five locations for barley and three wheat cultivating locations. F. graminearum loads reached levels of potentially serious threat for the commercial production of malting raw materials in both cereals (up to 29.1%). On the other hand, the mean percentage of kernels infected with F. culmorum was low to medium (up to 6.1%). The fungal invasions for years and locations were affected by meteorologic and other environmental factors and the pattern seemed to be consistent with species specific optimal conditions reported by other authors. PMID- 16255140 TI - Subtypes of genotype A Candida albicans isolates determined by restriction endonuclease and sequence analyses. AB - Systemic yeast infections are the Leading cause of mortality and morbidity in immunocompromized patients. Candida albicans, being the most frequently isolated fungal pathogen in these patients, can be divided into three genotypes (genotypes A, B and C) by 25S intron analysis. In our study, we found that molecular sizes of genotype A C. albicans isolates were heterogeneous. In order to determine the molecular basis of this difference, Haelll digestion was applied, and strains forming different band patterns were analyzed by automated sequence analysis. As a result of sequence analysis, eight different subtypes (a --> h) were found among genotype A C. albicans strains and an easy differentiation scheme consisting of Haelll and MspI digestions was constructed. PMID- 16255141 TI - High temperature-induced changes in exopolysaccharides, lipopolysaccharides and protein profile of heat-resistant mutants of Rhizobium sp. (Cajanus). AB - A thermosensitive wild-type strain (PP201) of Rhizobium sp. (Cajanus) and its 14 heat-resistant mutants were characterized biochemically with regard to their cell surface (exopolysaccharides (EPSs) and lipopolysaccharides (LPSs)) properties and protein profile. Differences were observed between the parent strain and the mutants in all these parameters under high temperature conditions. At normal temperature (30 degrees C), only half of the mutant strains produced higher amounts of EPSs than the parent strain, but at 43 degrees C, all the mutants produced higher quantities of EPS. The LPS electrophoretic pattern of the parent strain PP201 and the heat-resistant mutants was almost identical at 30 degrees C. At 43 degrees C, the parent strain did not produce LPS but the mutants produced both kinds of LPSs. The protein electrophoretic pattern showed that the parent strain PP201 formed very few proteins at high temperature, whereas the mutants formed additional new proteins. A heat shock protein (Hsp) of 63-74 kDa was overproduced in all mutant strains. PMID- 16255142 TI - Mycelial pellet formation by Penicillium ochrochloron species due to exposure to pyrene. AB - Five indigenous fungal strains with characteristics of the genus Penicillium capable of degrading and utilizing pyrene, as sole carbon source were isolated from soil of a former gas work site. Two strains were identified as Penicillium ochrochloron. One of the strains was able to degrade a maximum of 75% of 50 mg l( 1) pyrene at 22 degrees C during 28 days of incubation. The presence of pyrene in the medium resulted in an aggregation of hyphae into pellets by the two Penicillium ochrochloron strains. Formation of pellets was observed after 48 h of incubation with difference in size and texture between the two strains. This indicated the individual variation within the same genus of fungi. However, remaining strains did not show this behavior even though they were capable of utilizing pyrene as sole carbon source. The macro- and microscopic morphology of fungal pellets was studied using scanning electron microscopy. It was found that the addition of varying concentration of pyrene ranging from 10 to 50 mg l(-1) in the medium influenced shape and structure of the mycelial pellets. A two-fold increase in hyphal branching (with concomitant decrease in the average hyphal growth unit) was observed at a concentration of 10mg l(-1). The relevance of fungal growth and morphology for bioremediation of polycyclic aromatic hydrocarbons (PAHs) contaminated sites are discussed. PMID- 16255143 TI - In situ characterization of mercury-resistant growth-promoting fluorescent pseudomonads. AB - Pseudomonas fluorescens strains PRS9 and GRS1 (wild type) were made mercury resistant PRS9Hg(r) (147 microM HgCl2) and GRS1Hg(r) (55 microM HgCl2), respectively, in King's medium by enrichment selection and their in situ root colonization studies were carried out. Mercury resistant mutant of PRS9 was stable and resulted in significant increase in root and shoot fresh weight (P < 0.05). Both the mutants are positive for indoleacetic acid (IAA), 'P' solubilization and siderophore production. PRS9, potent 'P' solubilizer, exhibited higher 'P' solubilization as compared to GRS1. After 2 weeks of inoculation, the population level of wild type PRS9 and its mercury resistant mutants has increased (50 fold). Mercury resistance has no adverse effect on the growth promoting properties of mutants besides being comparable in its morphological and physiological properties with their wild type counterpart. Furthermore, mercury resistant character facilitates rhizospheric competition and thus helpful for establishment of growth promoting strains where metal ions are either limiting and/or present at toxic level. PMID- 16255144 TI - CgDN24: a gene involved in hyphal development in the fungal phytopathogen Colletotrichum gloeosporioides. AB - A cDNA corresponding to a transcript induced in culture by N starvation, was identified in Colletotrichum gloeosporioides by a differential hybridisation strategy. The cDNA comprised 905 bp and predicted a 215 aa protein; the gene encoding the cDNA was termed CgDN24. No function for CgDN24 could be predicted by database homology searches using the cDNA sequence and no homologues were found in the sequenced fungal genomes. Transcripts of CgDN24 were detected in infected leaves of Stylosanthes guianensis at stages of infection that corresponded with symptom development. The CgDN24 gene was disrupted by homologous recombination and this led to reduced radial growth rates and the production of hyphae with a hyperbranching phenotype. Normal sporulation was observed, and following conidial inoculation of S. guianensis, normal disease development was obtained. These results demonstrate that CgDN24 is necessary for normal hyphal development in axenic culture but dispensable for phytopathogenicity. PMID- 16255145 TI - N-butanoyl-L-homoserine lactone (BHL) deficient Pseudomonas aeruginosa isolates from an intensive care unit. AB - Acylated homoserine lactones (AHLs) are self-generated diffusible signal molecules that mediate population density dependent gene expression (quorum sensing) in a variety of Gram-negative bacteria, and several virulence genes of human pathogens are known to be controlled by AHLs. In this study, strains of Pseudomonas aeruginosa, Acinetobacter baumannii, Escherichia coli and Klebsiella pneumoniae, isolated from intensive care patients, were screened for AHL production by using AHL responsive indicator strains of Chromobacterium violaceum CV026 and Agrobacterium tumefaciens NT1. Positive reactions were recorded for all 50 isolates of P. aeruginosa and 10 isolates of Acinetobacter baumannii with Agrobacterium tumefaciens NT1. Surprisingly, most P. aeruginosa isolates gave negative results with C. violaceum CV026 in contrast to previous reports. This suggests that the new isolates of P. aeruginosa either failed to make short chain AHLs or the level of the signal molecule is very low. PMID- 16255146 TI - ArcA works with Fnr as a positive regulator of Vitreoscilla (bacterial) hemoglobin gene expression in Escherichia coli. AB - Low oxygen induction of the bacterial (Vitreoscilla) hemoglobin gene (vgb) by the Arc system was investigated, as the presumptive vgb Crp site was found to have 73% identity to the Escherichia coli consensus ArcA site. The role of ArcA by itself and with Fnr was examined in E. coli using the wild type vgb promoter and promoter mutants affecting the Fnr and Crp (presumptive ArcA) sites and E. coli strains with all combinations of fnr+/fnr-, arcA+/arcA- genotypes. High-level transcription required both ArcA and Fnr systems to be functional; low oxygen induction required at least one of ArcA and Fnr to be intact. Levels of Vitreoscilla hemoglobin protein (VHb) followed the same trends as seen with mRNA, although the relative decreases in the mutants relative to wild type were less than with transcription. Growth of cells was stimulated by VHb, generally to a greater extent as VHb levels increased. PMID- 16255147 TI - Identification of homologues to the pathogenicity factor Pat-1, a putative serine protease of Clavibacter michiganensis subsp. michiganensis. AB - Hybridization of Clavibacter michiganensis subsp. michiganensis total DNA against the pathogenicity gene pat-1 indicated the presence of pat-1 homologous nucleotide sequences on the chromosome and on plasmid pCM2. Isolation of the corresponding DNA fragments and nucleotide sequence determination showed that there are three pat-1 homologous genes: chpA (chromosome) and phpA and phpB (plasmid pCM2). The gene products share common characteristics, i.e. a signal sequence for Sec-dependent secretion, a serine protease motif, and six cysteine residues at conserved positions. Gene chpA located on the chromosome is a pseudogene since it contains a translational stop codon after 97 of 280 amino acids. In contrast to pat-1, cloning of the plasmid encoded homologs phpA and phpB into the avirulent plasmid free Cmm strain CMM100 did not result in a virulent phenotype. So far, no proteolytic activity could be demonstrated for Pat 1, however, site specific mutagenesis of pat-1 showed that the serine residue in the motif GDSGG is required for the virulent phenotype of pat-1 and thus Pat-1 could be a functional protease. PMID- 16255148 TI - Growth and siderophore production of Xylella fastidiosa under iron-limited conditions. AB - In this study, the production of siderophores by Xylella fastidiosa from the citrus bacteria isolate 31b9a5c (FAPESP - ONSA, Brazil) was investigated. The preliminary evidence supporting the existence of siderophore in X. fastidiosa was found during the evaluation of sequencing data generated in our lab using the BLAST-X tool, which indicated putative open reading frames (ORFs) associated with iron-binding proteins. In an iron-limited medium siderophores were detected in the supernatant of X. fastidiosa cultures. The endophytic bacterium Methylobacterium extorquens was also evaluated. Capillary electrophoresis was used to separate putative siderophores produced by X. fastidiosa. The bacterial culture supernatants of X. fastidiosa were identified negative for hydroxamate and catechol and positive for M. extorquens that secreted hydroxamate-type siderophores. PMID- 16255149 TI - Standards of care for adults with convulsive status epilepticus: Belgian consensus recommendations. AB - Status epilepticus (SE) is a significant health problem, affecting approximately 1,000 to 4,000 individuals per year in Belgium. A workshop was convened by a panel of neurologists from major Belgian centers to review the latest information relating to the definition, diagnosis and treatment of convulsive SE. The panelists sought to make recommendations for practising neurologists, but also primary care physicians and physicians in intensive care units when initiating emergency measures for patients with convulsive SE. As there is an association between prolonged seizures and a poor outcome, the importance of early (within the first 5 minutes of seizure onset) and aggressive treatment is to be stressed. In addition to general systemic support (airway, circulation), intravenous administration of the benzodiazepines lorazepam or diazepam is recommended as first-line therapy. Intramuscular midazolam may also be used. If SE persists, second-line drugs include phenytoin or valproate, and third-line drugs the barbiturate phenobarbital, the benzodiazepine midazolam, or the anaesthetics thiopental or propofol, or eventually ketamine. If the patient does not recover after therapy, monitoring of seizures should involve an electroencephalogram to avoid overlooking persistence of clinically silent SE. As a general rule, the intensity of the treatment should reflect the risk to the patient from SE, and drugs likely to depress respiration and blood pressure should initially be avoided. If initial treatment with a benzodiazepine fails to control seizures, the patient must be referred to the emergency unit and a neurologist should be contacted immediately. PMID- 16255150 TI - Clinical contribution of PET neurotransmission imaging in neurological disorders. AB - Imaging neurotransmission in vivo using positron emission tomography (PET) is a rapidly expanding clinical science. The present review summarizes the actual contribution of PET imaging to clinical problems in movement and seizure disorders and dementia. PMID- 16255151 TI - The role of chemotherapy in the treatment of low-grade glioma. A review of the literature. AB - Low-grade gliomas (LGG) are a group of uncommon neuroglial tumors of the central nervous system. They are characterized by a grade I or II according to the WHO classification. Grade I tumors are non-invasive and amenable to surgical resection with curative intent. Diffuse infiltrating LGG (WHO grade II) are tumors with a highly variable prognosis. Curative resection can only rarely be achieved and progression is characterized by transformation into a high-grade glioma (WHO grade III-IV). There are only limited evidence-based treatment recommendations for the management of progressive LGG because of a lack of data from prospective randomized trials. Most often radiotherapy is offered to patients with symptomatic and/or progressive disease. Three randomized trials have failed to demonstrate a survival improvement with either early versus delayed radiation or with a higher dose of radiation. The potential role of chemotherapy for the treatment of LGG has only been addressed in phase II trials. The PCV-chemotherapy regimen is associated with considerable toxicity that limits its applicability. The results with temozolomide (TMZ) chemotherapy have been more promising. Patients with chemosensitive LGG as predicted by heterozygotic loss of chromosomal arms Ip and 19q or methylation of the promoter of the MGMT gene in the genome of the glioma cells respond to TMZ. Radiotherapy will be compared to chemotherapy asfirst line treatment for LGG in two phase III studies that are planned for by the brain tumor group of the European Organization for Research and Treatment of Cancer (BTG-EORTC) and Radiation Therapy Oncology Group (RTOG). PMID- 16255152 TI - Neuroprotection in acute ischemic stroke. AB - Neuroprotection of patients with acute ischemic stroke should start at the scene and continue in the ambulance with the assessment and treatment of the airway, breathing, circulation, body temperature, and blood glucose. The key goal in eligible patients should be fast vessel recanalization with intravenous recombinant tissue-type plasminogen activator Results from a meta-analysis suggest that systemic thrombolysis is effective when given within 4.5 hours after stroke onset. The time window extends to 6 hours for patients undergoing intravascular thrombolysis. Acute stroke patients should be admitted to stroke care units. A crucial component of neuroprotection is the prevention of secondary brain damage, which can be caused by hypoxemia, hypotension, hyperthermia and hyperglycemia. This can be achieved by avoiding complications, e.g. aspiration, and intensive control of oxygenation, hydration and blood pressure, body temperature, blood glucose, and cardiac monitoring. Neuroprotective agents are designed to try to salvage brain tissue within the penumbra. Thus far, despite promising preclinical studies, clinical trials with neuroprotective drugs in acute ischemic stroke have been disappointing. However, we have been able to identify many of the factors that were responsible for these failures, and better designed clinical trials with neuroprotective drugs should look more promising. Mild induced hypothermia is another form of neuroprotective treatment that is currently being investigated in acute stroke. PMID- 16255153 TI - A putative generalized model of the effects and mechanism of action of high frequency electrical stimulation of the central nervous system. AB - High-frequency stimulation (HFS) of neural structures has been used since 1997 as an alternative to lesions in functional neurosurgery of movement disorders, and more recently, it has been applied to the treatment of epilepsies, obsessive compulsive disorders, cluster headaches, and has other applications in experimental models, particularly for obesity. Although their clinical efficacy is not questioned, and that the effects most of the time parallel those of ablative techniques, leading to the concept of functional inhibition, the intimate mechanisms by which HFS induces excitation within fiber bundles and seems to inhibit cellular nuclei is still strongly debated. Principally due to the observation of long-term clinical effects over a period up to 15 years, it is clear that the mechanism is not due to a progressive lesion, as at every moment the interruption of stimulation reverses totally the effects. There is no current proof that long-term HFS is able to reset neural networks, or to induce profound modifications of the functional organization or of the synaptic connectivity. To understand what is responsible for the immediate, reversible and adaptable effects of HFS, several mechanisms must be considered, which might be involved simultaneously or in sequence: i) Jamming of neural transmission through stimulated nuclei is one possibility, based on the principle that the regular imposed activity might drive the neurons to fire in a regular pattern, making it impossible to transmit more subtle messages, either normal or abnormal. Although it is difficult to prove this type of mechanism, it might account for the reports of increased activity following HFS in various structures. ii) Direct inhibition of spike initiation at the level of the membrane could be due to activation of inhibitory terminals, particularly gaba-ergic, or by a blockade of the voltage gated ion channels. iii) Recent data show that HFS decreases the production and release of low molecular weight proteic neurotransmitters, which could account for the functional inhibition while the efferent axon is still excited by the electrical stimulus. iv) Retrograde activation of upstream neuronal structures, as reported in the external pallidum during stimulation of STN, might be responsible of additional jamming-like effects due to collisions with descending spikes. PMID- 16255154 TI - Cell therapy for neurological disorders: a comprehensive review. AB - Neurodegenerative diseases are characterized by the irreversible loss of neurons involved in networks, important for specific physiological functions. At present, several renewable cell sources stand in line to replace fetal brain cells as potential cell source for transplantation in the damaged brain. Recent developments raise the hope that selective populations of different neuronal phenotypes could be made "on demand". However, for every potential cell source there are still a lot of questions and drawbacks, which need to be resolved before a cell source could become the standard for clinical neuronal transplantation. The recent finding that the brain responds to damage by increased endogenous neurogenesis could prelude new "neurothrophic therapies", based on stimulating this endogenous repair. From preclinical studies it is evident that different disease mechanisms require different cell therapy approaches, depending on the underlying factor of the disease, the identity of neuronal systems that are involved and the complexity of networks that are affected. In this review the potential of different cell sources, including the endogenous progenitor cells, are discussed. Also results of preclinical and clinical transplantation studies in three different disease models are critically evaluated. PMID- 16255155 TI - Effect of the rectus femoris motor branch block on post-stroke stiff-legged gait. AB - Six hemiparetic stroke patients presenting with a stiff-legged gait underwent a motor branch block of rectus femoris (RF). A gait analysis, including synchronous kinematic, dynamic, energetic recordings was performed before and after motor branch block of RF. The electrical activity of RF, Vastus Medialis, Vastus Lateralis, Vastus Intermedius was also recorded. Before nerve block, the sagittal kinematic data showed a decrease in both the mean maximum knee angular displacement during swing phase and the mean angular knee velocity at toe-off. In the sagittal kinetic data, the mean knee extension moment was increased. The mean mechanical work, internal and external work, was increased and the mean energy cost was also decreased. After the motor branch block of Rectus Femoris, the sagittal mean knee angular displacement and the mean internal work of the gait were significantly improved. PMID- 16255156 TI - Metastatic germ cell testicular tumor of the eye and brain: case report. AB - The aim of this report is to contribute to the clinical understanding of this rare combined pathology. Intraocular metastatic tumors are rarely encountered pathologies. Mostly encountered primary sites are breast, prostate, kidney, lung and skin. Testis as a primary site for orbital metastases is rather rare. Metastasis to orbit usually occurs to periorbital structures, but to vitreus is relatively rare. Here we describe a case of intraocular metastatic tumor to the vitreus originating from testicular embryonal cell carcinoma. Patient with intraocular tumor had also intracranial metastasis. He was operated on for intracranial lesion, after an uneventfull postoperative period he was discharged. 2 months later when he died, postmortem examination of the intraocular lesion reported as metastatic embryonal cell carcinoma. As a conclusion testicular embryonal cell carcinoma, eventhough rare, must be considered to metastasize to the eye. PMID- 16255157 TI - Large simple trials in psychiatry: providing reliable answers to important clinical questions. PMID- 16255158 TI - RCTs in psychiatry: challenges and the future. PMID- 16255159 TI - Practical clinical trials for schizophrenia. PMID- 16255160 TI - Schizophrenia, social class and immigrant status: the epidemiological evidence. AB - BACKGROUND: By the mid-1960s, the importance of socio-economic status for schizophrenia had been demonstrated in terms of differences between social-class groups in prevalence and incidence rates, illness course and outcome, and treatment experience. In the causation-selection debate, however, opinion had swung in favour of the selection hypothesis. AIMS: To reassess evidence on the social-class distribution of schizophrenia in Britain, and to compare this body of research with population-based studies of schizophrenia risk in socially disadvantaged ethnic minorities. METHOD: Systematic review of medical and psychological data-bases. RESULTS: Epidemiological research, while confirming the importance of premorbid social decline, has also provided support for the environmental 'breeder' hypothesis. High psychosis rates have been confirmed in ethnic minorities; in particular among Afro-Caribbean and other Black immigrants whose low social status cannot be accounted for by selective downward social drift or segregation. CONCLUSIONS: There are striking parallels, both in the epidemiology of schizophrenia and in social characteristics, between the lower class indigenous groups highlighted by earlier psychiatric surveys and African Caribbean populations in Britain's inner cities today. These similarities underline the need for a broader perspective in the search for environmental risk factors. PMID- 16255161 TI - [Interferon-alpha-induced psychiatric side effects in patients with chronic viral hepatitis: a prospective, observational, controlled study]. AB - AIMS: Patients with chronic viral hepatitis suffer from a high prevalence of psychiatric problems. Furthermore, the treatment for chronic viral hepatitis, with interferon (IFN) alpha, induces the occurrence of further psychopathological symptoms. The authors examined whether patients with a pre-existing psychiatric diagnosis had more severe IFN alpha-induced psychiatric adverse effects, and whether they were more likely to interrupt the IFN alpha therapy, compared with control patients with no pre-existing psychiatric diagnosis. They also examined the psychopharmacological management of the interferon-alpha-induced psychiatric side effects. METHODS: The authors studied prospectively 60 patients with chronic hepatitis B or C in Cagliari, Italy. Patients underwent psychiatric assessment before starting interferon alpha and monthly throughout the therapy. RESULTS: After adjusting for the baseline psychopathology, there was no statistically significant difference in interferon-alpha-induced psychiatric adverse effects between patients with a pre-existing psychiatric diagnosis and controls. There was also no evidence that psychiatric cases were more likely than controls to interrupt the IFN alpha therapy because of psychiatric side effects. Moreover, there was no difference in the psychiatric adverse effects severe enough to require psychopharmacological treatment. Finally, psychopharmacological management successfully treated psychiatric symptoms induced by the IFN alpha. CONCLUSIONS: Patients with a pre-existing psychiatric diagnosis do not have a specific vulnerability to interferon-alpha-induced psychiatric adverse effects. PMID- 16255162 TI - [Community attitudes towards mental illness and socio-demographic characteristics: an Italian study]. AB - AIM: To assess the association between socio-demographic characteristics and community attitudes towards mentally ill people. METHOD: We assessed a sample of 280 subjects, stratified for sex and age, which has identified using the electoral registers of Brescia. A letter was sent to everyone in order to introduce the future potential study participant to the topics of the public attitudes towards mental illness and it included an invitation to take part in the study. After, 280 subjects were contacted by telephone. Finally, 174 persons, who expressed their willingness to collaborate, were visited by a team of four trained interviewers. The instruments used were: a semi-structured interview; the Community Attitudes to the Mentally Ill (CAMI) inventory, which is composed by 40 statements, concerning the degree of acceptance of mental health services and mentally ill patients in the community; and the Fear and Behavioural Intentions (FABI) inventory, which is composed by 10 items, concerning fears and behavioural intentions towards mentally ill people. 106 subjects refused to participate. RESULTS: Factor analysis of the CAMI revealed three components Physical distance and fear, Social isolation and Social responsibility and tolerance. Factor 1 is associated with: people >61 years old; people being divorced/widowed/living separated; people who haven't participated in social or volunteer activities. Factor 2 is associated with: people > 41 years old; people being schooled at a level that's higher than elementary level; unemployed people. Factor 3 doesn't present any associations. CONCLUSIONS: The results of this study outline the need to: a) promote interventions focused to improve the general attitude towards people with mental illness; b) to favour specific actions in order to prevent or eliminate prejudices in subgroups of the population. PMID- 16255163 TI - Empathy in general practice consultations: a qualitative analysis. AB - AIMS: Empathy is an important basis of all provider-patient interaction, including general practice. Little has been known about the details of how general practitioners express empathy to their patients. In the present study, it was a main aim to carry out a detailed qualitative study of actual sequences through which empathy was achieved, by focusing on the form, function, and categorisation of such sequences. METHODS: The study drew on the qualitative method of conversation analysis. Transcriptions of 77 British general practitioner-patient interactions were analysed in detail in order to identify and characterise sequences in which empathy was achieved. RESULTS: Four types of empathy were identified, described, and analysed: cognitive empathy, affective empathy, sharing empathy, and nurturant empathy. Although sequences involving empathy typically were brief and occurred relatively rarely, the analyses demonstrated that the sequences often concerned matters of great importance to the interactants. CONCLUSIONS: The study underlined the need to focus also on phenomena that occur relatively rarely in consultations. For this purpose, the method of conversation analysis was a useful tool. PMID- 16255164 TI - [A survey on the prejudice and the stereotypes of mental illness in two communities with or without psychiatric Residential Facilities]. AB - AIMS: Description of prejudice and stereotypes of mental illness in two samples with or without psychiatric Residential Facilities in their communities. It has been evaluated if the ongoing naturalistic relationship with the mentally ill was sufficient to modify the stigma of mental illness. METHODS: The study has been carried out in 7 small towns with Residential Facilities and in 7 small towns without Residential Facilities on a sample of 560 people. Each respondent was asked to fill the Questionnaire on the Opinions about Mental Illness-General Population's version (QO-GP). RESULTS: Data on 557 respondents (99.5%) were collected. The most frequent opinion in the general sample was to reject the possibility to admit patients to Psychiatric Hospital. The significance (p < .01) between the two samples was found for only 5 items that concern the "treatment outcome", while no difference was found about the stereotypes of mental illness. CONCLUSIONS: The results outline that the ongoing naturalistic relationship do not modify the stereotypes of mental illness, but only some aspects of prejudice (care and civil rights). As a consequence we need to plan structured, effective and more specific educational campaigns on mental illnesses. PMID- 16255165 TI - Isolation of cDNA and enzymatic properties of betaine aldehyde dehydrogenase from Zoysia tenuifolia. AB - We isolated cDNAs encoding betaine aldehyde dehydrogenase (BADH, EC 1.2.1.8) from the salt-tolerant Poaceae, Zoysia tenuifolia by polymerase chain reactions. Zoysia betaine aldehyde dehydrogenase 1 (ZBD1) is 1892bp long and codes for 507 amino acids. The deduced amino acid sequence of ZBD1 is 88% similar to the sequence of rice BADH. Ten cDNA clones were isolated from a cDNA Library of salt treated Z. tenuifolia by using the ZBD1 fragment as a probe. The proteins coded in some clones were more homologous to BBD2, the cytosolic BADH of barley, than to ZBD1. To investigate their enzymatic properties, ZBD1 and spinach BADH were expressed in Escherichia coli and purified. The optimal pH of ZBD1 was 9.5, which was more alkaline than that of spinach BADH. ZBD1 was less tolerant to NaCl than spinach BADH. ZBD1 showed not only BADH activity but also aminoaldehyde dehydrogenase activity. The Km values of ZBD1 for betaine aldehyde, 4 aminobutyraldehyde (AB-ald), and 3-aminopropionaldehyde (AP-ald) were 291, 49, and 4.0 microM, respectively. ZBD1 showed higher specific activities for AB-ald and AP-ald than did spinach BADH. PMID- 16255166 TI - Modulation of carbonic anhydrase activity in two nitrogen fixing cyanobacteria, Nostoc calcicola and Anabaena sp. AB - The activity of enzyme carbonic anhydrase (CA) was investigated in two diazotrophic cyanobacteria, Anabaena sp. (ARM 629) and Nostoc calcicola, in the presence of CO2/NaHCO3 and different inhibitors. The CA activity increased when the cells were pretreated with a high concentration of CO2/NaHCO3 and then transferred to ambient level CO2. Maximum activity of CA was observed after 8 h of incubation in light on transfer of cells from high Ci to ambient level CO2, and was low when incubated in dark. Addition of the photosynthetic inhibitor DCMU brought about a differential reduction in CA activity, depending on the carbon source (NaHCO3/CO2). CA inhibitors--ethoxyzolamide (EZ) and acetazolamide (AZ)- inhibited the enzyme activity in both the genera, but the extent of inhibition was greater in Anabaena sp. than in N. calcicola. Such a variation in extent of inhibition/stimulation of CA activity being different in the two genera reflects differences in their inherent potential and genetic background. The relevance of such cyanobacterial strains as CO2 sinks is also discussed. PMID- 16255167 TI - Modulation of phosphoenolpyruvate carboxylase in vivo by Ca2+ in Amaranthus hypochondriacus, a NAD-ME type C4 plant: possible involvement of Ca2+ in up regulation of PEPC-protein kinase in vivo. AB - The properties of phosphoenolpyruvate carboxylase (PEPC) were studied, with respect to calcium (Ca2+), in leaves of Amaranthus hypochondriacus, a C4 plant. Experiments were conducted in vitro (by adding Ca2+ during enzyme assay) or in vivo (by feeding Ca2+ to intact leaves through petiole). Inclusion of 10 microM Ca2+ during assay marginally increased (<30%) malate sensitivity of PEPC in extracts from dark-adapted leaves. The effect of Ca2+ was marginal on PEPC in extracts from illuminated leaves. Upon applying a low concentration of Ca2+ to leaves, the PEPC activity in leaves increased by 1.5-fold, while inhibition by malate decreased markedly. The light activation of PEPC in Ca2+-fed leaves was slightly higher than in the absence of Ca2+-ethyleneglycol-bis-(beta-aminoethyl ether) N,N,N',N'-tetra acetic acid (EGTA). To assess further the role of Ca2+, 5 mM EGTA (Ca2+ chelator) was either added during the enzyme assay or fed to leaves through petiole. EGTA had no effect on PEPC, when added during enzyme assay. Upon feeding EGTA, the PEPC activity in the dark-adapted leaf extracts increased by 30%, and the effect on malate sensitivity was marginal. However, there was a decrease in PEPC activity in illuminated extracts, resulting in a marked decrease in the extent of light activation of PEPC. The extent of phosphorylation of PEPC was much higher in Ca2+ or Ca2+-EGTA-fed leaves than in the control, but EGTA decreased the light-induced phosphorylation. Our results suggest that optimal alone concentration of Ca2+ is essential for PEPC in leaves of A. hypochondriacus, particularly in vivo. We suggest that Ca2+ regulates PEPC, at an upstream level, such as transcription, by modulating PEPC-protein kinase, thus facilitating the light activation of PEPC. PMID- 16255168 TI - Influence of host cultivars and Rhizobium species on the growth and symbiotic performance of Phaseolus vulgaris under salt stress. AB - In order to study the effect of salt stress on the Rhizobium-common bean symbiosis, we investigated the response of both partners, separately and in symbiosis. The comparison of the behaviour of five cultivars of Phaseolus vulgaris differing in seed colour, growing on nitrates and different concentrations of NaCl, showed genotypic variation with respect to salt tolerance. Coco Blanc was the most sensitive cultivar, whereas SMV 29-21 was the most tolerant one. At the Rhizobium level, two strains previously selected for their salt tolerance were used: Rhizobium tropici strain RP163 and Rhizobium giardinii strain RP161. Their relative growth was moderately decreased at 250mM NaCl, but they were able to grow at a low rate in the presence of 342 mM NaCl. Their viability at the minimal inhibitory concentration was slightly affected. The effect of salinity on Rhizobium-plant association was studied by using the tolerant variety SMV 29-21 and the sensitive one Coco Blanc inoculated separately with both strains. In the absence of salinity, the strains induced a significantly higher number of nodules on the roots of the cultivar SMV 29-21 compared to those of Coco Blanc. Concerning effectiveness, both strains were similarly effective with SMV 29-21, but not with Coco Blanc. In the presence of salinity, Coco Blanc was more severely affected when associated with RP163 than with RP161. Salinity affected the nodulation development more than it affected the infection steps. Neither of the two strains was able to nodulate SMV 29-21 under saline conditions, in spite of the fact that this was considered the most salt-tolerant variety. The unsuccessful nodulation of SMV 29-21 could be related to the inhibition by salt of one or more steps of the early events of the infection process. In conclusion, N-fixing plants were found to be more sensitive to salt stress than those depending on mineral nitrogen. Evidence presented here suggests that a best symbiotic N2 fixation under salinity conditions could be achieved if both symbiotic partners, as well as the different steps of their interaction (early events, nodule formation, activity, etc.), are all tolerant to this stress. PMID- 16255169 TI - Hydrogen peroxide pre-treatment induces salt-stress acclimation in maize plants. AB - The effect of exogenously applied H2O2 on salt stress acclimation was studied with regard to plant growth, lipid peroxidation, and activity of antioxidative enzymes in leaves and roots of a salt-sensitive maize genotype. Pre-treatment by addition of 1 microM H2O2 to the hydroponic solution for 2 days induced an increase in salt tolerance during subsequent exposure to salt stress. This was evidenced by plant growth, lipid peroxidation and antioxidative enzymes measurements. In both leaves and roots the variations in lipid peroxidation and antioxidative enzymes (superoxide dismutase, ascorbate peroxidase, guaiacol peroxidase, glutathione reductase, and catalase) activities of both acclimated and unacclimated plants, suggest that differences in the antioxidative enzyme activities may, at least in part, explain the increased tolerance of acclimated plants to salt stress, and that H2O2 metabolism is involved as signal in the processes of maize salt acclimation. PMID- 16255170 TI - Higher plant-like fluorescence induction and thermoluminescence characteristics in cyanobacterium, Spirulina mutant defective in PQH2 oxidation by cytb6/f complex. AB - Characterization of the photosynthetic electron transport in a mutant of Spirulina platensis, generated by chemical mutagenesis, demonstrated that the electron transfer from the plastoquinone (PQ) to cytochrome b6/f was slowed. Thermoluminescence (TL) measurements suggested the presence of reversed energy flow via PQ, which resulted in an emergence of the plant-like after-glow TL band at 45 degrees C that could be enhanced by the transthylakoidal pH gradient and could be eliminated by an uncoupler, FCCP. The localization of the changes in the electron transport of the mutant cells measured by various methods revealed that the re-oxidation of the PQ pool is hampered in the mutant compared to the wild type cells. The reduction in energy migration was localized between PQ and PS I reaction centers. PMID- 16255171 TI - Cadmium effects on growth and mineral nutrition of two halophytes: Sesuvium portulacastrum and Mesembryanthemum crystallinum. AB - Growth, cadmium accumulation and potassium and calcium status were studied in two halophytes from Aizoaceae family: Sesuvium portulacastrum and Mesembryanthemum crystallinum. After multiplication, the seedlings were cultivated on nutrient solution supplemented with NaCl (100mM) and CdCl2 (0, 50, 100, 200 and 300 microM). After 1 month of treatment, plants were harvested and the dry weight, as well as the Cd, K and Ca concentrations in tissues were determined. Results showed that S. portulacastrum, a perennial halophyte with slow growth, is significantly more tolerant to Cd than M. crystallinum, an annual plant. Cd severely inhibited Mesembryanthemum growth even at the lowest Cd concentration in culture medium (50 microM), and did not modify significantly that of Sesuvium. For both halophytes, Cd accumulation was significantly higher in the roots than in the shoots. However, Cd concentration reached 350-700 microg g(-1) DM in the shoots, values characteristic of Cd hyperaccumulator plants. The addition of Cd in the culture medium led to a disturbance of Ca and especially K nutrition, suggesting the possibility to improve plant growth and Cd phytoextraction of both halophytes by increasing nutrient availability in the culture medium. PMID- 16255172 TI - Developmental expression of ASG- 1 during gametogenesis in apomictic guinea grass (Panicum maximum). AB - We have used Western blue-visualized in situ-hybridization (ISH) to monitor the expression of apomixis-specific gene-1 (ASG-1, GenBank accession number AB000809) during gametogenesis in obligate-sexual and facultative-apomictic (aposporic) genotypes of guinea grass (Panicum maximum). The in situ-analysis revealed that ASG-1 is not expressed in the ovule during early floral development in both, the facultative apomicts (A1 stage) and the obligate sexuals (S1 stage). With the appearance of the aposporous initial cell(s) in the ovule of the apomictic type (A2-1 stage), ASG-1 expression is strong and specific to this apomixis-specific cell. ASG-1 expression continued through different stages of aposporous embryo sac development (A2-2 stage), indicating that the gene may play a role in this developmental process. Regular embryo sacs in sexual types did not show hybridization signals (S2 stage). However, strong ASG-1 expression was detected in immature pollen grains and young embryos in both reproductive types, suggesting that ASG-1 may be an allele derived from the obligate-sexual wild type. Expression in pollen grains faded with maturation. In a heterologous system, using Paspalum notatum, a facultative-aposporic tropical grass (bahia grass), identical results were obtained. The results are discussed in view of the fact, that ASG-1 shows some homology to genes known to be seed- or embryo specific or involved in processes related to cell growth. PMID- 16255173 TI - Cloning and biochemical properties of CDPK gene OsCDPK14 from rice. AB - A rice CDPK gene, OsCDPK14 (AY144497), was cloned from developing caryopses of rice (Oryza sativa cv. Zhonghua 15). Its cDNA sequence (1922 bp) contains an ORF encoding a 514 amino acids protein (56.7kD, pl 5.18). OsCDPK14 shows the typical structural features of the CDPK family, including a conserved catalytic Ser/Thr kinase domain, an autoinhibitory domain and a CaM-like domain with four putative Ca2+-binding EF hands. Subcellular targeting indicated that OsCDPK14 was located in the cytoplasm, probably due to the absence of myristoylation and palmitoylation motifs. OsCDPK14 was expressed in Escherichia coli and purified from bacterial extracts. The recombinant protein was shown to be a functional protein kinase using Syntide-2, a synthetic peptide. Kinase activity was shown to be Ca2+-dependent, and this activation was strongly enhanced by Mn2+ and inhibited by W7 in vitro. These results provide significant insights into the regulation and biochemical properties of OsCDPK14, suggesting OsCDPK14 may be a signal factor of cytoplasm in rice plant. PMID- 16255175 TI - Production of mouse adiponectin, an anti-diabetic protein, in transgenic sweet potato plants. AB - Adiponectin is a 30kDa protein exclusively produced and secreted from adipocytes and as a cytokine has been found to link obesity, insulin resistance, and type 2 diabetes. Production of biologically active adiponectin in large scale is desirable for pharmaceutical applications. Mouse adiponectin cDNA was used for developing transgenic sweet potato plants via Agrobacterium-mediated transformation. The presence of the transgene was verified by PCR and DNA gel blot analysis. Further investigated were five independent transgenic lines, all of which expressed high levels of adiponectin mRNA. Immuno blot analysis with a mouse adiponectin antiserum revealed that, in addition to a 29 kDa-protein which co-migrates with the adiponectin protein produced in Escherichia coli cells, a 31 kDa-protein was produced, indicative of a post-translational modification of the protein. The transgenic plants did not show obvious differences in growth rate and morphology in response to adiponectin production. PMID- 16255174 TI - Molecular cloning, characterization and expression of a novel jasmonate-dependent defensin gene from Ginkgo biloba. AB - A novel defensin gene was isolated from Ginkgo biloba. The full-length cDNA of G. biloba defensin (designated as Gbd) was 534bp. The cDNA contained a 240-bp open reading frame encoding an 80-amino acid protein of 5.68 kDa with a potential 30 aa signal peptide. The putative GbD mature protein showed striking similarity to other plant defensins, representing low molecular size antimicrobial polypeptides. Eight cysteine sites conserved in plant defensins were also found in GbD at similar positions. Three-dimensional structure modeling showed that GbD strongly resembled defensin from tobacco (NaD1) and consisted of an alpha-helix and a triple-strand antiparallel beta-sheet that were stabilized by four intramolecular disulfide bonds, implying GbD may have functions similar to NaD1. The genomic DNA gel blot indicated that Gbd belonged to a multigene family. Expression analysis revealed that Gbd was up-regulated by wounding and methyl jasmonate treatments, suggesting that Gbd is potentially involved in plant resistance or tolerance to pathogens during wounding. PMID- 16255176 TI - Enhanced silymarin accumulation is related to calcium deprivation in cell suspension cultures of Silybum marianum (L.) Gaertn. AB - Elimination of calcium ions from the medium of cell cultures of Silybum marianum (L.) Gaertn increased flavonolignan production. Silymarin accumulation was not altered by treatment of cultures with the calcium ionophore A23187. The specific Ca2+ chelator, EGTA, enhanced the silymarin content in cells by 200%, and its secretion by 3-4 times. The inorganic ion La3+, as well as the calcium channel inhibitor verapamil, also stimulated production. Several reagents known to block intracellular calcium movement, such as ruthenium red, thapsigargin and TMB-8 appreciably increased silymarin accumulation. These results suggest that inhibition of external and internal calcium fluxes plays a significant role in flavonolignan metabolism of S. marianum cell cultures. PMID- 16255177 TI - Heavy metal stress reduces the deposition of calcium oxalate crystals in leaves of Phaseolus vulgaris. AB - Calcium oxalate (CaOx) crystals in plants may serve as a sink for the absorption of excess calcium, and they could play an important role in heavy metal detoxification. In this study, the effect of heavy metals and different calcium concentrations on the growth of calcium oxalate crystals in leaves of Phaseolus vulgaris was investigated. Different analytical techniques were used to determine the influence of exogenous lead and zinc on CaOx deposition and to detect a presence of these metals in CaOx crystals. We found a positive correlation between the calcium concentration in the nutrient medium and the production of calcium oxalate crystals in leaves of hydroponically grown plants. On the other hand, addition of the heavy metals to the nutrient medium decreased the number of crystals. Energy dispersive X-ray spectrometry did not detect the inclusion of heavy metals inside the CaOx crystals. Our investigation suggests that CaOx crystals do not play a major role in heavy metal detoxification in P. vulgaris but do play an important role in bulk calcium regulation. PMID- 16255179 TI - National PA week: more than just a week. PMID- 16255180 TI - In the beginning: a PA history roundtable. PMID- 16255178 TI - Nancy and the KIDney quilt. PMID- 16255181 TI - Urinary problems in the elderly adult. PMID- 16255182 TI - Detecting celiac disease in adult patients. PMID- 16255183 TI - Case report: for facial swelling, look below the belt. PMID- 16255184 TI - It's 8 am. Do you know who your gay and lesbian patients are? PMID- 16255186 TI - Knee pain and a limp after a fall. PMID- 16255185 TI - A 63-year-old woman reports new changes in a long-standing "rash". PMID- 16255187 TI - Case of the month. Septate uterine pregnancy. PMID- 16255188 TI - [Application of anticoagulants in patients after operations for gastro-intestinal bleeding]. AB - Our clinical experience witness the necessity of administration of direct anticoagulants in prophylactic dosage to patients, operated on for gastro intestinal bleeding. The most effective and secure is preparation of the low molecular weight fractionized heparins group--Fraxparin. The summarized data concerning group of patients, in whom Fraxyparin was applied, witness the absence of unfavourable peak of effect after the preparation injection, gradual action and holding on of the interinjection time plato, the absence of significant additive noncontrollable action of Fraxparin after each subsequent injection made. PMID- 16255189 TI - [Efficacy of antibiotic prophylaxis using zinacef in postoperative period]. AB - The results of intravenous injection of Zinacef for purulent--septic complications in patients with various acute surgical diseases of the abdominal cavity organs were presented. Main principles and rules of antibioticoprophylaxis and antibioticotherapy conduction in surgery were substantiated. PMID- 16255190 TI - [Application of the proton pump inhibitors in treatment of an acute gastro intestinal bleeding of the ulcer etiology]. AB - The results of treatment of 53 patients, suffering an acute gastro-intestinal bleeding of the ulcer etiology are adduced. The proton pump inhibitor Omez infusion have promoted the improvement of the patients treatment results. PMID- 16255191 TI - [Efficacy of endovascular prophylaxis of pulmonary thromboembolism]. AB - The improvement of the treatment results and efficacy of the endovascular prophylaxis methods in patients at risk for pulmonary thromboembolism (PTHE) occurrence constitute the aim of the investigation. The work is based on retrospective analysis of the treatment results of 114 patients, to whom cava filter (CF) was implanted for the PTHE accidents prophylaxis. During hospital period after the CF implantation the PTHE reattacks were revealed in 4 patients, the CF embolism and thrombosis--in 10, the ascending thrombosis--in 8, the CF dislocation and migration--in 8. Late results of the CF implantation were studied in 54 patients in terms from 1 to 10 years. In 3 of them the symptomless perforation of vena cava inferior (VCI) wall was diagnosed with nonsignificant CF diversion, distal migration of CF with adequate preservation of antiembolic function--in 1, the thrombosis had occurred in 5 patients. The CF implantation into VCI is an effective method of endovascular prophylaxis of fatal PTHE. The high efficacy of CF "Osot" was evidenced, witnessing the expediency of its clinical application. PMID- 16255192 TI - [Effect of neuroleptnarcosis on the change of hemodynamic parameters during tissues transplantation]. AB - Changes of the central and peripheral hemodynamics indexes during operation for the tissues purulent-trophic defects closure by free muscular--nerve--vascular flap, arising during neuroleptnarcosis and in immediate postoperative period, were discussed. PMID- 16255193 TI - [Results of treatment of polytrauma in pregnant women]. AB - There was studied the traumatic disease course in 11 pregnant women with different variants of severe combined thoracic and abdominal trauma, constituting 1.6% of total number of injured persons, suffering polytrauma. Polytrauma have had caused severe course of traumatic disease in majority of pregnant women. In the early postshock period the complications had occurred in 90.9% of injured persons. Antenatal death of fetus was noted in 81.8% of pregnant women with polytrauma. All pregnant women after polytrauma are alive. PMID- 16255194 TI - [Microsurgery of the lower extremities peripheral nerves damage]. AB - New possibilities of restoration microsurgery of the lower extremities peripheral nerves damage were presented, taking into account different conditions of regeneration of the operated nerve, basing on analysis of 242 clinical observations. PMID- 16255195 TI - [Dermal equivalent as an inductor of the lower extremities ulcer healing]. AB - There were adduced the results of application of the dermal equivalent, manufactured using the tissue engineering methods, in complex of treatment of the lower extremities trophic ulcer. The pathogenetic substantiation was suggested for application of the method in the treatment of trophic ulcers. Possible advantages of the dermal equivalent imposition vs autodermoplasty performance were estimated. PMID- 16255196 TI - [The plasm welding of the digestive system organs in experiment]. AB - The plasm welding of the gut organs--perspective trend, which permits to increase the surgeon possibilities in performance of operative intervention. The results of operations, performed on large intestine and stomach, trusts, that it is possible to apply the streams of the high temperature gas plasm for the tissues dissection and connection. There are such advantages of the method, as the operation time shortage, simplicity in manipulating "Plasmomed" apparatus, absence of foreign bodies in the connection zone, high degree of hemostasis achieved. After the operation there was observed rapid restoration of the main structure elements in the large intestine and stomach wall, localized in the plasm welding zone. There was elaborated and tested a new method of the living tissues welding, and the processes, occurring in tissues within the plasm welding application zone, were studied up. PMID- 16255197 TI - [Application of photochemically activated ointment based on polyethylene oxides for treatment of peritonitis]. AB - There was given evidence, using the complex of morphological, morphometrical, biochemical and microbiological investigations in experiment, of the peritonitis treatment efficacy applying photochemically activated laevomecol ointment. The experiment was performed in 120 adult male white mongrel rats, following the recommendations of Vancouver's convention for biomedical investigations. PMID- 16255198 TI - [Postoperative pancreatitis]. PMID- 16255199 TI - [Defects of the thumb development in association with other hand abnormalities and methods of their correction]. PMID- 16255200 TI - [Rare complication of gastric cancer]. PMID- 16255201 TI - [Rare observation of gastric neurofibroma]. PMID- 16255202 TI - [Intraintestinal hemorrhage from the processus vermicularis stump after appendectomy]. PMID- 16255203 TI - [The method of treatment of colostasis in elderly and senile patients]. PMID- 16255204 TI - [Rare complication of an acute cholecystitis]. PMID- 16255205 TI - [Miniinvasive methods of surgical treatment of pancreatic pseudocyst]. PMID- 16255206 TI - [Laparoscopic extirpation of splenic cyst]. PMID- 16255207 TI - [The remove of stomatological needle from bronchus BVII]. PMID- 16255208 TI - [Observation of venous aneurysm of orifice of the leg large subcutaneous vein, simulating incarcerated femoral hernia]. PMID- 16255209 TI - [Laparoscopic intervention for multiple trauma in conditions of the district hospital]. PMID- 16255210 TI - [Dirofilariasis as a surgical problem]. PMID- 16255211 TI - [The pioneer of clinical transplantology (to the 110th anniversary professor Yu. Yu. Voronoi]. PMID- 16255212 TI - [Subcardial resection for gastric cancer]. AB - The method of subtotal gastric resection for cancer, stipulating for lower esophageal sphincter and gastric cardia, was elaborated in the clinic. Good immediate results, low rate of postoperative complications, the absence of gastroesophageal anastomosis sutures insufficience together with significant improvement of functional results due to preservation of lower esophageal sphincter function permit to recommend subcardial gastric resection as a method of choice in a distal gastric third cancer as well as in the early gastric corpus cancer. PMID- 16255213 TI - [Formation of invaginational esophago-intestinal anastomosis in reconstructive extirpation of gastric stump]. AB - In 1986-2003 yrs the interventions for cancer of the operated stomach were performed in 32 patients. Reconstructive extirpation of gastric stump (REGS) was conducted in 31 patient, subtotal reresection of gastric stump according to Roux- in 1. Esophago-intestinal anastomosis (EIA) was formed using modified method of K. N. Tsatsanidi. Postoperative mortality was 3.1%. In 1994-2003 yrs, while performing REGS and gastrectomy, EIA was formed in 529 patients. The rate of complicated healing of EIA 1.3%. The insufficiency of the EIA sutures after REGS performance was absent. PMID- 16255214 TI - [Results of application of a new operative procedure in treatment of patients with nonspecific complete extrasphincteric rectal fistula]. AB - In 2003-2005 yrs in the division of proctology there were operated 52 patients for nonspecific complete extrasphincteric rectal fistula using an original method. There were analyzed immediate results of intervention of the method proposed and conventional procedure according to Ryzhikh (the closest analogue). It was established that application of a new operative method is highly effective, essentially influences the general results of treatment, promotes the lowering of the postoperative complications, recurrences and incontinence rate. PMID- 16255215 TI - [Peculiarities of splanchnic hemodynamics after performance of partial portosystemic shunting]. AB - Complex hemodynamical investigations were conducted in 1985-2004 yrs in 60 patients with hepatic diseases before the operation, in 6-8 and 12-24 months after performance of portosystemic shunting operation (in 26 patients was formed H-like mesentericocaval anastomosis, in 10--central splenorenal anastomosis, in 14--splenorenal anastomosis side-to-side, in 10--H-like splenorenal anastomosis). Performance of shunting operation had promoted the lowering of volumetric speed of blood flow and pressure in portal vein, her diameter reduction, the general hepatic blood flow lowering. The lowering of blood flow in the portal vein system after the shunting have caused enhancement of arterial hepatic blood flow. PMID- 16255217 TI - [Analysis of immediate complications and mortality after performance of pancreatic resection]. AB - Experience of performance of pancreatic resection during two years in the clinic was summarized. In 90.4% of patients there was revealed malignant tumor, as a rule, in III-IV stage, complicated by jaundice, gastrointestinal bleeding and ileus. Postoperative complications rate was 28.8%, mortality--9.6%, after proximal pancreatic resection--11.4%. Pancreatic resection, including en bloc procedure, may be performed in the patients relatively securely, with not high complications rate and mortality. PMID- 16255216 TI - [Complex treatment of patients with cholangiogenic hepatic abscess]. AB - Results of treatment of 47 patients with cholangiogenic hepatic abscess were analyzed. Clinical, laboratory and special methods of investigation were applied for diagnosis. The authors consider ultrasound investigation (USI), computer tomography and the abscess puncture under the USI control with subsequent cytological and bacterial control the decisive methods in diagnosis of hepatic abscess. In complex of treatment of patients miniinvasive technologies were applied--the abscess puncture, its cavity drainage under USI control transcutaneus transhepatic cholangiostomy, endoscopic papillosphincterotomy with lithotripsy and nasobiliary drainage, according to indications. Efficacy of the abscess cavity sanation, using miramistinum and decasan, was proved. In general therapy the directed transport of medicines was applied. PMID- 16255218 TI - [Albumin-mediated peritoneal dialysis as method of restorational therapy in hepatic insufficiency]. AB - New method of extracorporeal detoxication of organism, which is applied for temporary restoration of hepatic detoxication function in patients with hepatic insufficiency, albumin-mediated peritoneal dialysis, was presented. The main principle of this technology is a transfer through highly permeable dialysis membrane of toxins, which are in affinity with albumin, from the blood to acceptor. The donor's human albumin, circulating in the closed contour, serves as acceptor. The watersoluble lowmolecular substances are excreted according to the concentration gradient. For rapid restoration of acceptoral ability of the donor's albumin solution it was subjected to hemodialysis and carboperfusion. The performance of the procedure guarantees excretion of toxins, combined with albumin, along with excretion of watersoluble toxins. PMID- 16255219 TI - [Intracavitary application of silics for the treatment of inflammatory-purulent processes]. AB - The results of clinical application of Silics--new sorbent of polydirected action, based on highly disperse amorphous silica, were presented. Silics owes unique properties, which are manifested while its intracavitary application for treatment of inflammatory-purulent processes. Osmotic activity of sorbent is more than 500%, waterabsorbability--up to 1000%, it sorbates up to 800 mg/g of proteins, microorganisms--up to 10(10) of the colony forming units on 1 g. Silics promotes lysis of necrotized tissues. In clinical application of Silics in 146 patients with retroperitoneal space phlegmon in an acute destructive pancreatitis, in 23--with abdominal cavity abscess, in 34--with inflammatory purulent pleuropulmonary diseases it was established, that this preparation promotes the intoxication severity lowering, the reduction of microbal sowing of purulent cavities and resistance of the wound microflora to antibotics. The securing of fragmentation of necrotized tissues and the fibrin deposits, lowering of the exudate viscosity rises efficacy of the inflammatory focus drainage. Application of Silics for the treatment of inflammatory-purulent processes permits to improve essentially its results on the account of lowering of mortality and the treatment cost, and the shortage of the patients stationary treatment. PMID- 16255221 TI - [Morphological changes in the lungs and characteristics of thromboemboli in patients with thromboembolism of the pulmonary artery]. AB - Basing on results of pathologoanatomical investigations, performed in 26 cases, the lungs changes in pulmonary thromboembolism were analyzed. It was shown, that an acute thromboembolism of a. pulmonalis main branches did not cause an acute necrosis of pulmonary alveolar tissue, but induces progressive diffuse changes of their microcircular bed, concomitant with pulmonary hypertension. In cases of chronical pulmonary thromboembolism the diffuse vascular sclerosis of the nonaffected lung and the alveolar tissue emphysema are formed, and in such cases the signs of pulmonary hypertension showing hypertrophy of the muscular vessels middle layer as well as their intima hyperplasia are revealed. This determines the expediency of earlier start of thrombolytic therapy conduction as well as surgical intervention in an acute pulmonary thromboembolism. PMID- 16255220 TI - [Immunotherapy: the strategic approach to the treatment of patients with severe sepsis]. AB - Expediency of inclusion of immunoglobulins for intravenous infusion into algorhythm of intensive therapy in patients with severe sepsis was analyzed. PMID- 16255222 TI - [The role of computer tomography in differential diagnosis of benign and malignant tumors of the adrenals cortex]. AB - Possibilities of computer tomography (CT) application for differential diagnosis of tumors of the adrenal cortex (TAC) were studied. CT was conducted in 157 patients with TAC, including 75--with malignant (MTAC), 82--with benign (BTAC), permitting to revealed MTAC in 76% patients. In 17.1% patients with BTAC false positive results were noted, reducing, thus, the method diagnostic value. While the trustworthy signs of malignancy are absent, it is necessary to conduct morphological verification of the adrenal tumors nature using puncture biopsy and/or intraoperative express-cytological investigation. PMID- 16255223 TI - [Experience of application of echoosteometry in the face middle zone fracture]. AB - The method of diagnosis of the face skull middle zone fracture using comparative measurement of the ultrasound passage time between the face bones points, of the same name, but concerning the affected and noneffected sides, was proposed. In cases, where this index exceeds by 20% the subperiosteum fracture is diagnosed and in by more than 40%--the face bones fracture with dislocation. PMID- 16255224 TI - [Application of videothoracoscopy in surgical treatment of the anterior mediastinum diseases]. AB - In 1994-2000 yrs there were observed 248 patients with various mediastinal diseases, including 105--with primary diagnosis: mediastinal tumor. In 102 (41.8%) patients operative intervention was performed using videothoracoscopy method, in 146--conventional methods: left-sided partial oblique sternotomy, longitudinal and longitudinal-transversal sternotomy, thoracotomy. Indications and contraindications for performance of operations on the anterior mediastinum organs, using videothoracoscopy, were determined for the first time. PMID- 16255225 TI - [Biochemical indexes of blood in the cholelithiasis on the background of fatty liver]. AB - There was conducted the analysis of biochemical indexes in 45 patients suffering the cholelithiasis on the background of adipose hepatosis. In majority of them there were noted the lipids metabolism and hepatic function disorders, as well as activation of processes of peroxidal oxidation of lipids. PMID- 16255227 TI - [Endovascular intervention for pseudoaneurysm of the common carotid artery using stent-graft]. PMID- 16255226 TI - [The diseases of the epiploic appendages of the cecum appendix]. PMID- 16255229 TI - [Perspectives of using phototherapeutical apparatus Bioptron in medicine: an interview with professor K.A.Samoilova by S.Stevanovich]. PMID- 16255228 TI - [The treatment of hernial disease complicated with peritonitis in conditions of the district hospital]. PMID- 16255230 TI - Safety technologies for laparoscopic monopolar electrosurgery; devices for managing burn risks. AB - The use of monopolar electrosurgery during laparoscopy has many benefits. But it also creates the risk that high-frequency leakage currents could travel from the active electrode to patient tissue, causing burns. In some cases, these burns can be serious and even lead to life-threatening complications. In this Evaluation, we review four products designed to reduce the risk of leakage-current burns during these procedures. One of the products is an active-electrode shaft with a brightly colored inner layer of insulation, making it easier to spot insulation breaks that could allow leakage currents. Two of the products use high DC voltage to detect full-thickness breaks in electrode insulation. The other product is an active-electrode shielding system that provides protection during a procedure by conducting potentially dangerous leakage current away from the patient. We rate one product Preferred and the others Acceptable. However, we don't believe that any of these technologies are essential to safe monopolar laparoscopic electrosurgery. If these technologies are used, they should be viewed only as supplemental safety measures, not as a substitute for the general safe practices required during these procedures. PMID- 16255231 TI - Higher currents, greater risks: preventing patient burns at the return-electrode site during high-current electrosurgical procedures. AB - Patient burns at the site of the return electrode are a well-known, and generally well-managed, risk of electrosurgery. However, the use of newer electrosurgical devices and techniques that apply higher currents to the patient for longer periods of time has created a new set of burn risks. And the safety measures that have proven effective during conventional electrosurgery can not necessarily be relied on to prevent return-electrode-site burns during these high-current, long activation-time electrosurgical procedures. In this article, we explain the hazards and detail the factors that can increase the risks of return-electrode site injuries for a given procedure. We also provide recommendations to help facilities and clinicians minimize those risks. PMID- 16255232 TI - Return-electrode-site burns associated with Rita Medical Systems Model 1500 and 1500X radio-frequency generators. PMID- 16255233 TI - Skin burns resulting from the use of conductive distention/irrigation media during electrosurgery with a rollerablation electrode. PMID- 16255234 TI - Improper return-electrode selection and placement contribute to patient burn with the Smith & Nephew Vulcan ElectroThermal Arthroscopy System (EAS). PMID- 16255235 TI - VIP infant ventilator seriously injures newborn. PMID- 16255236 TI - Making friends while on the road. PMID- 16255237 TI - Meet the travelers. Jennifer Kuntz. PMID- 16255238 TI - Take the hassle out of packing. PMID- 16255239 TI - Unlocking the secrets to a good phone interview. PMID- 16255240 TI - Meet the travelers. Ingrid Ostenaa. PMID- 16255241 TI - Tobacco transcription factor WRKY1 is phosphorylated by the MAP kinase SIPK and mediates HR-like cell death in tobacco. AB - The salicylic acid-induced protein kinase (SIPK) of tobacco, which is a mitogen activated protein kinase (MAPK), is activated by various biotic and abiotic treatments. Overexpression of SIPK has been shown to trigger cell death. In this study, a targeted yeast two-hybrid approach identified the tobacco transcription factor WRKY1 as a potential substrate. SIPK phosphorylated WRKY1, which resulted in enhanced DNA-binding activity of WRKY1 to its cognate binding site, a W box sequence from the tobacco chitinase gene CHN50. SIPK-mediated enhancement of WRKY1 DNA-binding activity was inhibited by staurosporine, a general kinase inhibitor. Co-expression of SIPK and WRKY1 in Nicotiana benthamiana led to more rapid cell death than expression of SIPK alone, suggesting that WRKY1 is involved in the formation of hypersensitive response-like cell death and may be a component of the signaling cascade downstream of SIPK. PMID- 16255242 TI - Two classes of highly similar coiled coil-nucleotide binding-leucine rich repeat genes isolated from the Rps1-k locus encode Phytophthora resistance in soybean. AB - A series of single genes protect soybean from the root and stem disease caused by the oomycete pathogen Phytophthora sojae. In the last two decades, Rps1-k has been the most stable and widely used Phytophthora resistance gene for the major soybean-producing regions of the United States. Four highly similar genes encoding coiled coil-nucleotide binding-leucine rich repeat (CC-NB-LRR)-type proteins were isolated from the Rps1-k locus. These genes were grouped into two classes based on their sequence identity. Class I contains three genes with identical open reading frames (ORF) and 5' end regions. Two of these genes were also identical at the 3' untranslated regions; the third gene showed a recombination breakpoint in the 3' untranslated region resulting in the combination of 3' end sequences of members from both classes. Reverse transcription-polymerase chain reaction analyses suggested that members of both classes of genes are transcribed at low levels. Representative members from each gene class were expressed in transgenic soybean plants. Analyses of independent R0, R1, R2, and R3 progeny populations suggested that both gene classes confer Phytophthora resistance in soybean. A possible evolutionary mechanism for the Class I gene family is proposed. PMID- 16255244 TI - Natural variation in the Arabidopsis response to the avirulence gene hopPsyA uncouples the hypersensitive response from disease resistance. AB - The plant hypersensitive response (HR) is tightly associated with gene-for-gene resistance and has been proposed to function in containing pathogens at the invasion site. This tight association has made it difficult to unequivocally evaluate the importance of HR for plant disease resistance. Here, hopPsyA from Pseudomonas syringae pv. syringae 61 is identified as a new avirulence gene for Arabidopsis that triggers resistance in the absence of macroscopic HR. Resistance to P. syringae pv. tomato DC3000 expressing hopPsyA was EDS1-dependent and NDR1 independent. Intriguingly, several Arabidopsis accessions were resistant to DC3000(hopPsyA) in the absence of HR. This is comparable to the Arabidopsis response to avrRps4, but it is shown that hopPsyA does not signal through RPS4. In a cross between two hopPsyA-resistant accessions that differ in their HR response, the HR segregated as a recessive phenotype regulated by a single locus. This locus, HED1 (HR regulator in EDS1 pathway), is proposed to encode a protein whose activity can cause suppression of the EDS1-dependent HR signaling pathway. HED1-regulated symptomless gene-for-gene resistance responses may explain some cases of Arabidopsis resistance to bacteria that are classified as nonhost resistance. PMID- 16255243 TI - A new hypothesis on the strategy for acquisition of phosphorus in arbuscular mycorrhiza: up-regulation of secreted acid phosphatase gene in the host plant. AB - The mycorrhiza-responsive phosphatase of Tagetes patula in symbiosis with Glomus etunicatum was detected by electrophoresis, was purified by column chromatography, and was characterized as acid phosphatase that was secreted into rhizosphere. The N-terminal amino acid sequence was determined by a gas-phase sequencer, and a cDNA fragment of the phosphatase gene (TpPAP1) was amplified by degenerate primers designed based on the N-terminal amino acid sequence. The full length cDNA was obtained by the rapid amplification of cDNA ends technique. The TpPAP1 was of host origin, and the cDNA was 1,843 bp long with a predicted open reading frame of polypeptide of 466 amino acids. Phylogenetic analysis revealed that the gene fell into the cluster of plant high-molecular-weight purple acid phosphatase. Expression analysis of the TpPAP1 in T. patula in symbiosis with Archaeospora leptoticha showed that the levels of transcripts increased eightfold by mycorrhizal colonization. Western blot analysis revealed that the 57-kDa protein corresponding to the mycorrhiza-responsive phosphatase increased by mycorrhizal colonization. The present study proposes a new strategy for acquisition of P in arbuscular mycorrhizal associations in which the fungal partner activates a part of the low-P adaptation system of the plant partner, phosphatase secretion, and improves the overall efficiency of P uptake. PMID- 16255245 TI - Role of methylotrophy during symbiosis between Methylobacterium nodulans and Crotalaria podocarpa. AB - Some rare leguminous plants of the genus Crotalaria are specifically nodulated by the methylotrophic bacterium Methylobacterium nodulans. In this study, the expression and role of bacterial methylotrophy were investigated during symbiosis between M. nodulans, strain ORS 2060T, and its host legume, Crotalaria podocarpa. Using lacZ fusion to the mxaF gene, we showed that the methylotroph genes are expressed in the root nodules, suggesting methylotrophic activity during symbiosis. In addition, loss of the bacterial methylotrophic function significantly affected plant development. Indeed, inoculation of M. nodulans nonmethylotroph mutants in C. podocarpa decreased the total root nodule number per plant up to 60%, decreased the whole-plant nitrogen fixation capacity up to 42%, and reduced the total dry plant biomass up to 46% compared with the wild type strain. In contrast, inoculation of the legume C. podocarpa with nonmethylotrophic mutants complemented with functional mxa genes restored the symbiotic wild phenotype. These results demonstrate the key role of methylotrophy during symbiosis between M. nodulans and C. podocarpa. PMID- 16255246 TI - Suppression of root nodule formation by artificial expression of the TrEnodDR1 (coat protein of White clover cryptic virus 1) gene in Lotus japonicus. AB - TrEnodDR1 (Trifolium repens early nodulin downregulation 1) encodes a coat protein of White clover cryptic virus 1. Its expression in white clover was down regulated at the time when root nodules formed. We surmised that its artificial expression would interfere with root nodulation. Therefore, we investigated the effects of its artificial expression on the growth and root nodulation of Lotus japonicus (a model legume). Transformants were prepared by Agrobacterium spp. mediated transformation. The growth of transformants was reduced and the number of root nodules per unit root length was greatly decreased relative to control. The concentration of endogenous abscisic acid (ABA), which controls nodulation, increased in plants containing TrEnodDR1. These phenotypes clearly were canceled by treatment with abamine, a specific inhibitor of ABA biosynthesis. The increase in endogenous ABA concentration explained the reduced stomatal aperture and the deformation of root hairs in response to inoculation of transgenic L. japonicus with Mesorhizobium loti. Transcriptome comparison between TrEnodDR1 transformants and control plants showed clearly enhanced expression levels of various defense response genes in transformants. These findings suggest that TrEnodDR1 suppresses nodulation by increasing the endogenous ABA concentration, perhaps by activating the plant's innate immune response. This is the first report of the suppression of nodulation by the artificial expression of a virus coat protein gene. PMID- 16255247 TI - Aspergillus infection inhibits the expression of peanut 13S-HPODE-forming seed lipoxygenases. AB - Oxylipins recently have been implicated as signaling molecules for cross-kingdom communication in plant-pathogen interactions. Linoleic acid and its two plant lipoxygenase (LOX) oxylipin products 9- and 13-hydroperoxy fatty acids (9S- and 13S-HPODE) have been shown to have a significant effect on differentiation processes in the mycotoxigenic seed pathogens Aspergillus spp. Whereas both fatty acids promote sporulation, 9S-HPODE stimulates and 13S-HPODE inhibits mycotoxin production. Additionally, Aspergillus flavus infection of seed promotes linoleate 9-LOX expression and 9S-HPODE accumulation. Here, we describe the characterization of two peanut seed lipoxygenase alleles (PnLOX2 and PnLOX3) highly expressed in mature seed. PnLOX2 and PnLOX3 both are 13S-HPODE producers (linoleate 13-LOX) and, in contrast to previously characterized 9-LOX or mixed function LOX genes, are repressed between 5-fold and 250-fold over the course of A. flavus infection. The results of these studies suggest that 9S-HPODE and 13S HPODE molecules act as putative susceptibility and resistance factors respectively, in Aspergillus seed-aflatoxin interactions. PMID- 16255248 TI - Identification of a tomatinase in the tomato-pathogenic actinomycete Clavibacter michiganensis subsp. michiganensis NCPPB382. AB - The insertion site of a transposon mutant of Clavibacter michiganensis subsp. michiganensis NCPPB382 was cloned and found to be located in the gene tomA encoding a member of the glycosyl hydrolase family 10. The intact gene was obtained from a cosmid library of C. michiganensis subsp. michiganensis. The deduced protein TomA (543 amino acids, 58 kDa) contains a predicted signal peptide and two domains, the N-terminal catalytic domain and a C-terminal fibronectin III-like domain. The closest well-characterized relatives of TomA were tomatinases from fungi involved in the detoxification of the tomato saponin alpha-tomatine which acts as a growth inhibitor. Growth inhibition of C. michiganensis subsp. michiganensis by alpha-tomatine was stronger in the tomA mutants than in the wild type. Tomatinase activity assayed by deglycosylation of alpha-tomatine to tomatidine was demonstrated in concentrated culture supernatants of C. michiganensis subsp. michiganensis. No activity was found with the tomA mutants. However, neither the transposon mutant nor a second mutant constructed by gene disruption was affected in virulence on the tomato cv. Moneymaker. PMID- 16255250 TI - Potato homologs of Arabidopsis thaliana genes functional in defense signaling- identification, genetic mapping, and molecular cloning. AB - Defense against pests and pathogens is a fundamental process controlled by similar molecular mechanisms in all flowering plants. Using Arabidopsis thaliana as a model, steps of the signal transduction pathways that link pathogen recognition to defense activation have been identified and corresponding genes have been characterized. Defense signaling (DS) genes are functional candidates for controlling natural quantitative variation of resistance to plant pathogens. Nineteen Arabidopsis genes operating in defense signaling cascades were selected. Solanaceae EST (expressed sequence tag) databases were employed to identify the closest homologs of potato (Solanum tuberosum). Sixteen novel DS potato homologs were positioned on the molecular maps. Five DS homologs mapped close to known quantitative resistance loci (QRL) against the oomycete Phytophthora infestans causing late blight and the bacterium Erwinia carotovora subsp. atroseptica causing blackleg of stems and tuber soft rot. The five genes are positional candidates for QRL and are highly sequence related to Arabidopsis genes AtSGT1b, AtPAD4, and AtAOS. Full-length complementary DNA and genomic sequences were obtained for potato genes StSGT1, StPAD4, and StEDS1, the latter being a putative interactor of StPAD4. Our results form the basis for further studies on the contributions of these candidate genes to natural variation of potato disease resistance. PMID- 16255249 TI - RNA interference of dual oxidase in the plant nematode Meloidogyne incognita. AB - RNA interference (RNAi) is a powerful tool for the analysis of gene function in model organisms such as the nematode Caenorhabditis elegans. Recent demonstrations of RNAi in plant parasitic nematodes provide a stimulus to explore the potential of using RNAi to investigate disruption of gene function in Meloidogyne incognita, one of the most important nematode pests of global agriculture. We have used RNAi to examine the importance of dual oxidases (peroxidase and NADPH oxidase), a class of enzyme associated with extracellular matrix cross-linking in C. elegans. RNAi uptake by M. incognita juveniles is highly efficient. In planta infection data show that a single 4-h preinfection treatment with double-stranded RNA derived from the peroxidase region of a dual oxidase gene has effects on gene expression that are phenotypically observable 35 days postinfection. This RNAi effect results in a reduction in egg numbers at 35 days of up to 70%. The in vitro feeding strategy provides a powerful tool for identifying functionally important genes, including those that are potential targets for the development of new agrochemicals or transgenic resistance strategies. PMID- 16255251 TI - A patient-centred approach to research and development. PMID- 16255252 TI - Professionalizing postgraduate medical education: schools and academies. PMID- 16255253 TI - Histopathology training schools 4 years on. AB - The initial three pilot histopathology senior house officer schools have improved training efficiency and effectiveness. Additional schools have been established to develop a national network of histopathology training schools. Their success provides lessons for the organization and delivery of structured training in other medical specialties. PMID- 16255254 TI - IT innovation in histopathology recruitment, training and research. AB - The introduction of national recruitment and cohort training in histopathology in England has produced significant challenges and highlighted opportunities for development. Innovative information technology solutions have been deployed to facilitate recruitment and enhance education provision and research. PMID- 16255255 TI - The intensive training and assessment programme in histopathology. AB - A large number of experienced overseas graduates are attracted to the UK and form a significant proportion of applicants for training positions in histopathology. An individually tailored 3-month post can be used to evaluate the overseas doctor's abilities and assess his/her suitability for higher specialist training. PMID- 16255256 TI - Mental health law: compulsory treatment in a general medical setting. AB - Compulsory treatment of severe mental disorder can seem complicated even to psychiatrists. This article attempts to explain the current legal framework as it applies in common clinical situations in a general medical context. PMID- 16255257 TI - The levonorgestrel-releasing intrauterine system in modern gynaecology. AB - The levonorgestrel-releasing intrauterine system was initially developed for contraception but is now widely used for a variety of gynaecological conditions. Compliance can sometimes be hampered by troublesome side effects (principally breakthrough bleeding) but appropriate counselling can reduce unnecessary discontinuation. PMID- 16255258 TI - Clinical governance: what foundation doctors need to know. PMID- 16255259 TI - Headache. PMID- 16255260 TI - How to carry out pericardial aspiration. PMID- 16255261 TI - Interpretation of elbow and forearm radiographs. PMID- 16255262 TI - Injuries around the elbow. PMID- 16255263 TI - A narrow complex tachycardia. PMID- 16255264 TI - So you want to be ... a cardiologist. PMID- 16255265 TI - Hepatitis C: tackling the silent epidemic. AB - Chronic hepatitis C virus infection is responsible for a significant and growing burden on NHS services. It is one of the commonest causes of liver cirrhosis and hepatocellular carcinoma, and the leading indication for liver transplantation. Major advances have been made in treatment, which can eradicate the virus in more than 50% of patients and reduce complications, but progress is hampered by inadequate detection and access to treatment. PMID- 16255266 TI - Compartment syndromes. AB - Compartment syndrome occurs when pressure within a closed muscle compartment exceeds the perfusion pressure and results in muscle and nerve ischaemia. Two distinct conditions are recognized: acute and chronic (exertional) compartment syndromes. Differences in aetiology, pathophysiology and management are elaborated on in this article. PMID- 16255267 TI - Pseudosubluxation of the cervical spine. PMID- 16255269 TI - A tasty week in international and public health. PMID- 16255268 TI - How to organize a 'taster' week for foundation year 2 doctors. AB - This article describes the aim of short, focussed placements ('taster' experiences), and outlines some of the practicalities of organizing them. PMID- 16255270 TI - Air bag safety in road traffic accidents. PMID- 16255271 TI - Iatrogenic Creutzfeldt-Jakob disease presenting 24 years after human growth hormone administration. PMID- 16255272 TI - Animal welfare, human health. PMID- 16255273 TI - A case of claudication in a young man. PMID- 16255275 TI - Huge choice for pension investors. PMID- 16255274 TI - What does a raised troponin 1 level in the elderly tell us? PMID- 16255276 TI - Should 22G spinal needles for accessing the subarachnoid space be banned? PMID- 16255277 TI - Support for hurricane rescue efforts. PMID- 16255278 TI - Crohn's and Johne's disease. PMID- 16255279 TI - Animal welfare a recurring theme. PMID- 16255280 TI - Where there's a will, there's a way. . . Part 1--protecting the future of your estate. PMID- 16255281 TI - Rehabilitating wounded urban possums. PMID- 16255283 TI - Cutaneous lymphosarcoma in a stallion. AB - Multiple cutaneous lymphosarcomas were diagnosed in an 8-year-old Thoroughbred stallion presented for evaluation of lumps on its scrotum. Histological examination of skin biopsy samples showed a homogenous pattern of lymphoid tissue suggestive of a T-cell lymphosarcoma. Immuno-histochemical tests showed a positive reaction to Rabbit/Anti-Human T-Cell, CD3 antibodies confirming T-cell lymphosarcoma. The animal was not treated and was subsequently euthanased. PMID- 16255282 TI - Improving seizure control in dogs with refractory epilepsy using gabapentin as an adjunctive agent. AB - OBJECTIVE: To assess whether there is a change in seizure activity in dogs with refractory epilepsy that are receiving appropriate doses of phenobarbitone and/or potassium bromide, when gabapentin is added to the therapeutic regimen. DESIGN: A prospective study of 17 dogs with a refractory seizure disorder, 16 of which have idiopathic epilepsy. PROCEDURE: Patients were stabilised using phenobarbitone and/or potassium bromide to produce tolerable therapeutic serum concentrations and dosed additionally with gabapentin at 35 to 50 mg/kg/d (divided twice or three times daily) for 4 months. Owners recorded seizure activity and side effects during this period in a standardised diary. Patients underwent monthly physical examinations and venepuncture to assess selected serum biochemical analytes, as well as phenobarbitone and bromide concentrations. Patients were further monitored for long-term response to adjunctive gabapentin therapy. RESULTS: There was no significant decrease in the number of seizures over the study period for the entire cohort, however three dogs stopped seizuring completely. There was a significant increase in the number of patients who showed an increase in the interictal period (P > 0.001). Serum alkaline phosphatase activity and triglyceride concentrations were elevated at baseline. There were no significant changes in biochemical analytes during the course of the study period. Side effects observed initially on addition of gabapentin included sedation and hind limb ataxia. The former resolved spontaneously after a few days; the latter after a slight reduction in bromide dose. Long-term, a further two patients became seizure free and ten patients remained on gabapentin indefinitely. No long-term side effects have become apparent. CONCLUSION: Addition of gabapentin to phenobarbitone and/or potassium bromide increased the interictal period and shortened the post-seizure recovery in some canine epileptics. In some dogs, seizures were prevented completely, while in others there was an increase in interictal period. The short-half life of gabapentin has advantages for seizure control, however its present high cost may prohibit therapy in large dogs. PMID- 16255284 TI - Lymphoma(s) showing epitheliotropism and diffuse skeletal muscle involvement presenting as a polymyopathy in a young dog. AB - A 3-year-old dog presented with a severe polymyopathy and subsequently developed nodular skin lesions. The disease in this dog was caused by lymphoma, showing cutaneous epitheliotropism as well as infiltration of skeletal muscle in conjunction with polymyositis. PMID- 16255286 TI - Knowledge and attitudes of Australian veterinarians to animal abuse and human interpersonal violence. AB - A survey of Australian veterinarians was undertaken to assess their amount of knowledge about, and their attitudes towards animal abuse, human violence and the link between the two. Results from the 185 respondents to a questionnaire that was either mailed out or hand delivered revealed a wide variety of definitions and attitudes towards abuse, with the majority of veterinarians recognising the link between human and animal abuse. The overwhelming majority of veterinarians believed that they should intervene in some way when confronted with either animal or human abuse, although most felt ill-equipped to deal with suspected human abuse. Almost 20% of cases of animal abuse had associated suspected or known human abuse. It is suggested that veterinarians need more resources made available to them to be able to deal more effectively with these situations. PMID- 16255285 TI - Chemical immobilisation of wild banteng (Bos javanicus) in northern Australia using detomidine, tiletamine and zolazepam. PMID- 16255287 TI - Recent veterinary graduates over the last five decades: initial career experiences. AB - OBJECTIVE: To describe changes in the initial work experiences of veterinary graduates over the last 50 years. DESIGN: A questionnaire, sent by mail. PROCEDURE: A questionnaire seeking information on work experience as a recent graduate was sent to about 100 veterinarians who graduated in or about each of 1950, 1960, 1970, 1980, 1990 and 2000, and 68% responded. Data were entered onto an Excel spreadsheet, and analysed with the SAS System 8 for Windows. RESULTS: The first veterinary position for about half the graduates of 1950 and 1960 was in government service. Since then an increasing percentage have started in small animal practice; the rest in mixed practice. Many earlier graduates worked alone and for long hours, with little or no respite after hours. Since 1950, the average recent graduate in private practice has worked progressively fewer hours with less after hours duty, and has worked with progressively more other veterinarians. Cattle were almost half the cases seen by graduates of 1950 and 1960. Dogs and cats were about 20% of the cases seen by those graduates, but this percentage ncreased progressively and was 77% of the cases seen by the graduates of 2000. Distemper was the most common transmissible disease of dogs seen by graduates of 1950-1970, but by few since then. Graduates of 1980 saw parvovirus most commonly; those of 2000 saw parvovirus and kennel/canine cough in about equal numbers. Two other major changes in canine cases seen by recent graduates have involved skin conditions, which increased progressively between 1950 and 2000, and trauma, mainly involving vehicles, which decreased as dogs have been restrained more effectively. Fracture repair was the most common surgical procedure performed by recent graduates between 1950 and 1970, but with decreasing incidence of trauma this has been overtaken by the removal of lumps and repair of lacerations. Barbiturates were used extensively by recent graduates for both induction and maintenance of anaesthesia, but with increasing use of anaesthetic machines from 1960 onwards, halothane and more recently isoflurane have largely taken their place for the maintenance of anaesthesia. CONCLUSIONS: Over the last five decades, the average recent graduate has had progressively more opportunity for support and advice from other veterinarians, to work more sociable hours, and to work with a narrower range of species, especially dogs and cats. PMID- 16255288 TI - Determining the sensitivity of abattoir surveillance for ovine Johne's disease. AB - OBJECTIVE: The objective of this study was to determine the sensitivity of abattoir surveillance of intestinal tract lesions for detecting ovine Johne's disease (OJD) under normal meatwork conditions. DESIGN: The design of this study was a diagnostic test validation. The three OJD inspectors were the diagnostic test and follow-up histopathological examination was used for test validation. PROCEDURE: Approximately 1200 sheep were procured from known high prevalence OJD infected farms. The sheep viscera were tagged (numbered) and then examined as they were processed on the abattoir line by three experienced meat inspectors. Their observations were independently recorded on a cassette tape. Specified sections of viscera were prepared and subjected to histopathological examination and these results were compared with the inspector diagnoses. RESULTS: The sensitivity of abattoir inspection for OJD varied between inspectors from 53 percent to 87 percent. The specificity varied from 97 to 100 percent. It appeared that the level of sensitivity for detecting disease was higher in lines of sheep where the disease was more prevalent. It also appeared that formal training was an important aspect in ensuring a high level CONCLUSION: Abattoir surveillance is a very economical and rapid method of assessing the OJD status of sheep. On the basis of these results it is reasonable to suggest that abattoir surveillance has a sensitivity of approximately 70 percent. This technique is useful as an ancillary to other testing regimes for negative assurance programs where a sheep identification system is used. PMID- 16255289 TI - Preliminary observations on the prevalence of sheep shedding Mycobacterium avium subsp paratuberculosis after 3 years of a vaccination program for ovine Johne's disease. PMID- 16255290 TI - [What's new in muscular pathology]. PMID- 16255291 TI - [The steel stomach]. PMID- 16255292 TI - [Cirrhosis: new concepts and great therapeutic progresses]. PMID- 16255293 TI - [Epidemiology and natural history of cirrhosis]. AB - The cirrhosis prevalence in the whole French population can be estimated between 2000 and 3300 cases/million of individuals. Alcoholism, hepatitis C virus and non alcoholic fatty liver disease are the three most common causes of cirrhosis in France. About 40% of patients with cirrhosis have compensated cirrhosis and are asymptomatic over long period of 1 to 10 years. Decompensation of cirrhosis was considered when a patient first developed one of the mayor complications of the disease (ascites, jaundice, encephalopathy or gastrointestinal haemorrhage). After the first decompensation, the incidence of death is about 10% by year, in the absence of hepatic transplantation. The mortality of patients with cirrhosis and acute varicose bleeding has greatly decreased over the past 2 decades in concurrence with an early and combined use of pharmacological and endoscopic therapy, but hepatocellular carcinoma is the most frequent life threatening complication. PMID- 16255294 TI - [Predictive factors for progression to cirrhosis in chronic liver diseases]. AB - Chronic liver diseases complications generally occur when fibrosis progresses to cirrhosis. It is recognised that some patients do not progress while others develop significant fibrosis. Factors influencing the fibrosis progression rate are especially studied in chronic hepatitis C. Among many identified factors, the most important are those warranted a medical action such as alcohol consumption, obesity, other metabolic disorders and immunosuppression in case of HIV-HCV coinfection. Especially, in case of chronic liver disease, regular alcohol consumption should be avoided; overweight and metabolic disorders should be controlled. The control of liver damage aetiologies could decrease or even stop fibrosis progression. PMID- 16255295 TI - [Liver cirrhosis in adults: etiology and specific treatments]. AB - Cirrhosis is the result of chronic inflammation and of the progressive increase of fibrosis. In France, hepatitis C infection is the second cause of cirrhosis after alcohol abuse. The other causes of cirrhosis are: hepatitis B infection, genetic haemochromatosis, autoimmune hepatitis, primary biliary cirrhosis, drug induced cirrhosis, secondary biliary cirrhosis, Wilson's disease and al antitrypsin deficiency. Etiological treatment is based upon: abstinence in case of alcoholic cirrhosis, the combination of pegylated interferon alpha (PEG IFN) with ribavirin in case of C viral cirrhosis, the PEG IFN and the nucleoside analogs in case of B viral cause; corticosteroids and immunosuppressive drugs in case of autoimmune cirrhosis; venesections in case of genetic haemochromatosis and stopping the drug in case of drug-induced cirrhosis. The complications of cirrhosis such as ascites, oesophageal varices, bleeding, hepatic encephalopathy and hepatocellular carcinoma mainly explain the high rate of morbidity and mortality. Liver transplantation is the established therapy for decompensated liver disease of any etiology significantly changed the outcome of patients with advanced cirrhosis. PMID- 16255296 TI - [The irreversibility of cirrhosis: "the jostle dogma"]. PMID- 16255298 TI - [Cirrhosis in MEDLINE]. PMID- 16255297 TI - [Managing the complications of cirrhosis]. AB - The main complications of cirrhosis are gastrointestinal bleeding related to portal hypertension, ascites, spontaneous peritonitis, hepato-renal syndrome, encephalopathy and hepatocellular carcinoma. The apparition of these complications constitutes a major event for the patient. In addition to the etiological treatment, take charge of complications can improve the prognosis of the patients. PMID- 16255299 TI - [Patient handout. What is cirrhosis of the liver?]. PMID- 16255300 TI - [To err is human]. PMID- 16255301 TI - [National ranking examination 2005: No 5 to 9]. PMID- 16255302 TI - [Chronic obstructive pulmonary disease]. PMID- 16255303 TI - [Giant cell arteritis and polymyalgia rheumatica]. PMID- 16255304 TI - [Tuberculosis]. PMID- 16255305 TI - [Mitral regurgitation]. PMID- 16255306 TI - [Jaundice]. PMID- 16255307 TI - [Typhus: an ideologised disease]. PMID- 16255308 TI - HIV/AIDS: an African-American epidemic. PMID- 16255309 TI - African-American men and their reflections and thoughts on prostate cancer. AB - The purpose of this study was to assess African-American males' knowledge, attitudes, and beliefs regarding prostate cancer and early detection methods. This was a cross sectional study conducted with 67 (N = 67) African-American men to assess their knowledge, attitudes, and beliefs regarding prostate cancer and prostate cancer early detection methods Data collection occurred at several urban churches in the Washington, D.C., metropolitan area. Findings from this study suggest that 91% of the men did not consider prostate cancer screening embarrassing or painful; 86.9% agreed that their physician would think they should have a Digital Rectal Exam (DRE) and Prostate Specific Antigen (PSA) tests performed, and believed in the efficacy of early prostate cancer screening However, 30% of the men would not want to know if they had prostate cancer, and 53.2% did not believe they were likely to get prostate cancer in the future. The implications and results of this study support the need for ongoing prostate education and screening programs for African-American men. PMID- 16255310 TI - Perceived health status and health-promoting behaviors of African-American and White informal caregivers of impaired elders. AB - Caregiving and its consequences are major concerns for nurses. Many studies have examined health-promoting behaviors in general (Duffy, 1993; Walker, Volkan, Sechrist, & Pender, 1988), but few studies have explored health-promoting behaviors of caregivers. The purpose of this study was to examine the effects of age, gender, race, and length of caregiving on perceived health status and health promoting behaviors of African-American and White informal caregivers of impaired elders. Data from a larger study was used to examine a conceptual model linking perceived health status and health-promoting behavior. A sample of 136 (n = 136) African-American and 257 (n = 257) White caregivers was recruited from northeastern Ohio through random digit dialing and interviewed face to face. English-speaking caregivers were selected for study who provided unpaid assistance or care for a minimum of five hours a week to an impaired person 60 years of age or older living in the community. The Health-Promoting Behavior Questionnaire, perceived health status, and a demographic profile were used to measure the study variables. Data were analyzed using hierarchical multiple regression. Findings from this study suggest that the overall fit of the conceptual model was significant, and that moreover, the relationship found between perceived health status and health-promoting behaviors supported Pender's model of health promotion (Pender, 1987). The relationship between perceived health status and health-promoting behaviors has not been documented in prior studies of caregivers, or in studies comparing the two racial groups of African Americans and Whites. PMID- 16255311 TI - African-American caregivers' expectations of physicians: gaining insights into the key issues of caregivers' concerns. AB - This qualitative study was done to determine what expectations, if any, African American caregivers' have of physicians; what were the various causes of caregivers' distress, and what were the participants' perceived level of satisfaction or lack of satisfaction with the physician-patient-caregiver relationship The participants in this current study were either providing or had provided assistance in the activities of daily living of elders diagnosed with Alzheimer's or dementia. Focus group methodology was used and participants were recruited from Oklahoma County, Oklahoma. A total of 13 participants, whose mean age was 53.8 years of age and whose education was at least that of high school graduate, participated in the study. The majority of the caregivers were children who were caring for parents. The participants were asked three basic questions relating to their experiences or frustrations and the questions were followed by a 45-minute discussion to allow for further elaboration. The Geriatric Depression Scale (GDS), and the Zarit Burden Interview, combined with a demographic information form, was used to define the characteristics of the caregivers (Sheik, et al., 1986; Zarit, Reever & Bach-Peterson, 1980; Vitaliano, Russo, Yung, Becker, & Maiuro, 1991; National Center for Cost Containment, 1993). Findings from this study suggest that of the 13 participants only 2 could be classified as depressed. However, the caregivers expected information, referral for services and assistance in recognizing disease progression. These elements were sometimes lacking as well as not always having effctive caregiver-physician interaction. Greater attention by physicians to the needs, stressors, and expectations of African-American caregivers may improve the caregiver-physician interaction and may reduce caregiver stress. Further studies in this area can add to the sparsely available information. PMID- 16255312 TI - Uncovering homeless African-American women's knowledge of breast cancer and their use of breast cancer screening services. AB - This study explored the knowledge of breast cancer and the use of breast cancer screening services by homeless African-American women in an attempt to understand their use of cancer screening services and whether they understood the information about breast cancer and breast cancer screening. A qualitative study using a focus group format was employed to obtain a deeper understanding of the participants' experiences as they related to knowledge of breast cancer and their use of breast cancer screening services. The sample consisted of 25 (N = 25) African-American women who were currently living in a transient shelter for homeless women and children. Three, one-hour focus group sessions were conducted over a three-week period. Sessions were audio taped and transcribed for analysis. Content analysis was used to analyze the data. Findings from the study revealed categories that were associated with the women's personal experiences with cancer, their knowledge of breast cancer and experiences with the use of screening services, and their motivations as they related to breast cancer screenings. These categories can be useful in providing access to care and in addressing cancer education, screening behaviors, and program planning in homeless African-American women. PMID- 16255313 TI - Nursing contributions to the elimination of health disparities among African Americans: review and critique of a decade of research. AB - The excessive burden of disease experienced by African-Americans has long been described by authorities in the public, private, and professional sector as a national health concern. Several reports have been published in the peer-reviewed literature that describe the outcomes of nurse-directed studies aimed at addressing the factors associated with the disparities experienced by African Americans and these reports were also aimed toward the design of interventions to reduce and/or eliminate them. However, little is known about the scope, quality, and impact of these efforts relative to the promotion of health and the prevention of disease among African-Americans. This report presents the results of a review, analysis, and critique of reports of outcomes of nursing research aimed toward reducing health-related disparities among African-Americans. These reports were described in a selected group of African-American nursing organizations and journals dedicated to providing a forum for the discussion of issues focused on cultural diversity, transcultural nursing, and multicultural health care issues. Included among the journals were the Journal of the National Black Nurses Association, the Journal of the Association of Black Nursing Faculty, the Journal of Chi Eta Phi Sorority, the Journal of Cultural Diversity, the Journal of Transcultural Nursing, and the Journal of Multicultural Nursing and Health. Results of the review will be reported in three parts, the first of which was reported in an earlier edition (Journal of National Black Nurses Association, Volume 15, No. 1) and the second part of which is reported here. The third part will be reported in a future issue of the Journal of National Black Nurses Association. The results revealed that this body of nursing research provides the profession with a broad base of knowledge and insights. This knowledge is relative to the individual and familial impact of cardiovascular disease, cancer, diabetes, HIV/AIDS, mental health and mental illness, and Sickle Cell Disease within the African-American community. Knowledge and insights relative to the concerns and needs of childbearing, parenting the elderly caregivers and caregiving in African-American population groups is essential if health-care disparities are to be truly eliminated. This review, analysis, and critique also revealed several gaps and limitations within this body of nursing research. PMID- 16255314 TI - Colorectal cancer in African-Americans: addressing the need for further research and research utilization. AB - Research on colorectal cancer and screening has lagged behind when compared to other cancers such as breast and prostate cancer. The purpose of this paper is to provide a brief overview of colorectal cancer including risks, development of the cancer, and screening and early detection recommendations. A critical analysis of nurse-initiated research on colorectal cancer in African-Americans, published in nursing research-intensive journals, was conducted to further the understanding of the factors that influence these disparities. The findings indicated that this body of research is very sparse. Given this fact, nurses need to consider the types of research that are being conducted and where this research is published. Given the current focus on health disparities, it is time that nurses step into the forefront to lead the way in research and research utilization. It is crucial that we start to see real and significant decreases in colorectal cancer disparities in African-Americans. PMID- 16255315 TI - Prostate cancer among African-American males: understanding the current issues. AB - Prostate cancer affects African-American males within the United States in a disproportionate number compared to White males. African-American males are 1.7 times more likely to develop and 2-3 times more likely to die from prostate cancer than White males. Numerous reasons for this disparity exist, including low socioeconomic status, distrust, conflicting cultural beliefs, and past health care experiences. Controversies surrounding this topic and perhaps contributing to the disparity include cancer-screening recommendations, cancer-related myths, and potential prevention modalities. Nursing research must focus on cancer related issues among African-Americans to increase the awareness and knowledge of health-care professionals and the public to help decrease morbidity and mortality within African-Americans and other minority populations, and particularly among more vulnerable sections of at-risk minority populations. This article focuses on current issues related to African-American men and prostate health. PMID- 16255316 TI - Stress in health professions students: myth or reality? A review of the existing literature. AB - The purpose of this literature review was to determine the extent to which health profession students experience counterproductive stress and to examine the existing gaps in the literature concerning all health profession students and stress. The literature search process included using Eric, Education Abstracts, Psyc INFO, ABIN, WebSpirs Databases, FactSearch, Biological Sciences, International Pharmaceutical Abstracts, MEDLINE, Lexis-Nexis, CINAHL and Registry of Nursing Research. Key articles were identified that documented stress, the causes of stress or "stressors," and psychiatric symptoms as a result of counterproductive stress in the disciplines of medicine, dentistry, nursing, and allied health. Findings from this analysis of the literature indicated that the highest incidence of stress occurred in medical students followed by dental and then nursing students. There was, however, a dearth of studies related to stress in pharmacy students alone, despite the fact that pharmacology requires a rigorous educational process that is similar to that of the other health professions. While it may be argued that pharmacy students may not be experiencing the detrimental levels of stress that affect other medical students, a study by Henning Ey, & Shaw (1998) found a disproportionately high level of distress in pharmacy students. The Henning study also compared the stress levels of medical, dental, nursing, and pharmacy students. This literature review documents that health profession students suffer from high levels of stress during their training. A comprehensive list of "stressors" is delineated for the health education profession students. However, there appears to be a lack of emperical evidence with regards to stress and stressors in pharmacy students. Although it can be extrapolated that pharmacy students may be suffering from similar levels of stress, there is a definite need for research in pharmacy students specifically. PMID- 16255317 TI - Drinking locations prior to impaired driving among college students: implications for prevention. AB - Drinking and driving is perhaps the most serious problem associated with heavy drinking among college students in the United States. In this study, the authors examined drinking locations prior to impaired driving in a college student sample. They administered the Impaired Driving Assessment to 91 college students identified as high risk for drinking and driving. Participants reported an average of 7.98 (SD = 7.67) impaired driving episodes during the past 5 weeks. Using a random effects model, the authors found that location was a significant predictor of blood alcohol content (BAC), F(6,89.6) = 3.62, p = 0.0029. After drinking alcohol at a party, students' average estimated BAC prior to driving (geometric M = 0.089) was significantly greater than all other drinking locations (geometric M = 0.033). The findings of this study provide insight into drinking locations prior to impaired driving and can be used by college health practitioners to develop appropriate interventions to reduce the magnitude of this problem. PMID- 16255318 TI - Celebration intoxication: an evaluation of 21st birthday alcohol consumption. AB - The authors designed this study to evaluate the prevalence and magnitude of heavy drinking among college students in celebrating their 21st birthdays and the impact of a birthday card suggesting moderation. The authors randomly assigned subjects to receive or not receive the card approximately 1 week prior to their birthday. Approximately 1 week after turning 21, the authors sent surveys to all subjects. Results based on 164 returned surveys indicated that 90% consumed alcohol, 75% went to a bar, 61% reached a blood alcohol content (BAC) above the legal driving limit, and 23% reached a BAC above .25. Results were similar for men and women. Although subjects generally liked the birthday card, it had no impact on their drinking or celebration plans. Findings suggest the need for additional attention focusing on specific alcohol-related events and further development of prevention approaches that are event specific. PMID- 16255319 TI - Uncomplicated E. coli urinary tract infection in college women: a follow-up study of E. coli sensitivities to commonly prescribed antibiotics. AB - Treatment of uncomplicated urinary tract infections (UTIs) has changed in the past few years with researchers advocating empiric treatment for shorter periods of time without the use of cultures. Researchers report that antibiotic resistance of Escherichia coli (E. coli) to commonly prescribed antibiotics in uncomplicated UTIs has been increasing. Trimethoprim/sulfamethoxazole (TMP/SMX) is 1 of these antibiotics. Researchers also report that resistance patterns may differ depending on the geographic area of the United States. In this study, the authors present the results of a 7-month retrospective chart analysis of 98 E. coli sensitivities to commonly prescribed antibiotics in the treatment of uncomplicated UTIs at a college health service. They examined the more common antibiotic choices and analyzed their in vitro responses. Of these antibiotics, ciprofloxacin, nitrofurantoin, amoxicillin/clavulanate, and TMP/SMX had the highest sensitivity rates. The authors compared the results with a previous study that they performed at the same institution in 1993. The results of this study show a sensitivity rate of 86% for TMP/SMX. When compared with the previous result of 87%, this represented a 1% change. Because of this slight decrease in sensitivity and the increasing concern over resistance, the authors suggest that they will continue to reevaluate the resistance pattern in their population on a regular basis. This will help determine if there is a need for modifying choices of empiric therapy for UTIs. PMID- 16255320 TI - College students' motivation for physical activity: differentiating men's and women's motives for sport participation and exercise. AB - Despite the many clear benefits of an active lifestyle, lack of physical activity is a significant health problem in the college population. A key issue in physical activity research is developing an understanding of motivation. Although physical activity takes many forms, most research designed to enhance motivation for and adherence to physical activity focuses on exercise behavior and ignores sport participation. In this study, the authors compare motivations for sport participation versus exercise among college students. Results indicate that participants were more likely to report intrinsic motives, such as enjoyment and challenge, for engaging in sport, whereas motivations for exercise were more extrinsic and focused on appearance and weight and stress management. The findings suggest that motives for sport participation are more desirable than those for exercise and may facilitate improved adherence to physical activity recommendations. PMID- 16255321 TI - Weight-related issues and high-risk sexual behaviors among college students. AB - Individuals with high body mass index (BMI), negative body image, and unhealthy weight control behaviors may be more likely to engage in high-risk sexual behaviors than others. Researchers have not fully investigated these relationships in a sample of college students. The authors collected data regarding several weight-related issues and high-risk sexual behaviors (response rate = 39%) from 1,168 college students. Over half reported at least 1 high-risk sexual behavior. After the authors controlled for several demographic characteristics, they found that higher BMIs were associated with increased odds of having a casual sexual partner (odds ratio [OR] = 2.70) and being intoxicated at last intercourse (OR = 2.25). Among women, unhealthy weight control behaviors were associated with having a casual partner (OR = 2.12), using no or unreliable contraception (OR = 1.98), and being intoxicated at last intercourse (OR = 2.31). Weight-related issues were not associated with sexual behaviors among college men. The authors describe their findings within two health behavior frameworks, and they discuss implications for research and health education. PMID- 16255322 TI - Eating disorders in graduate students: exploring the SCOFF questionnaire as a simple screening tool. AB - The results of several studies have established the validity of the SCOFF questionnaire (a 5-question screening tool for eating disorders), but researchers need to explore further replicability using the US version in the graduate school population. In this study, the authors asked 335 graduate students attending the Northwestern student health clinic on the Chicago campus to complete a written survey anonymously. A total of 305 (91%) patients completed the survey. The sensitivity and specificity for the SCOFF was 53.3% and 93.2%, respectively. This produced a PPV (the proportion who tested positive on the screen and actually had an eating disorder) of 66.7% and an NPV (the proportion of those who tested negative on the screen and actually did not have an eating disorder) of 88.7%. More than 80% of respondents were dissatisfied with their shape and weight, with over one third having a moderate to severe body image disturbance. The SCOFF is an easy instrument to administer that health care providers can use to screen for eating disorders in the primary care setting. PMID- 16255323 TI - Predictors of smoking and smokeless tobacco use in college students: a preliminary study using web-based survey methodology. AB - Cigarette smoking and smokeless tobacco (SLT) use are associated with numerous health hazards and economic costs, and rates of tobacco use have recently increased among young adults. In this study, the authors compared predictors of smoking and SLT use among college students (N = 21,410) from 13 Texas universities using a Web-based survey. Results revealed that sex, belonging to a fraternity or sorority, participation in intercollegiate sports, peer influences, and ethnicity predicted smoking and SLT use. Although common factors predicted both lifetime and current smoking and SLT use, patterns of prediction differed across dependent variables. The authors discuss implications for developing tobacco prevention programs targeting specific risk factors salient to the young adult population. PMID- 16255324 TI - A meta-analysis of college students' physical activity behaviors. AB - The authors reviewed studies on college students' physical activity (PA) behaviors and found that previous research on this topic focused on describing college students' PA patterns and their determinants. Researchers reported that about 40% to 50% of college students are physically inactive. More important, health and PA professionals in higher education have not been able to effectively increase students' PA behaviors. Interventions to promote students' PA are still at an early stage and have only produced moderate effects. The authors found primary problems with the current research on this topic to be 3-fold: (1) college students' PA has been seriously neglected as a research topic, (2) there is a lack of multiple-level approaches (ie, personal, psychosocial, and environmental levels) for examining PA behaviors in the college student population, and (3) measures of PA are subjective and inconsistent, which makes comparisons of PA patterns among different samples very difficult or impossible. PMID- 16255325 TI - Reaction to "new meningococcal vaccine recommendations under consideration". PMID- 16255326 TI - Advising patients about patient safety: current initiatives risk shifting responsibility. AB - BACKGROUND: Many health care providers now disseminate advisories telling patients what they can do to avoid errors and harms in their care. METHODS: The content of five leading safety advisories for patients was analyzed and a critique of their development, content, and impact was developed, drawing on published literature and 40 interviews with a diverse sample of 50 key informants. FINDINGS: Very little is known about the effects of the distribution of safety advisories to patients, but several grounds for concern were identified. There was a lack of attention to patients' perspectives during the development of advisory messages, and the advisories say little about what health care providers should do to ensure patient safety. Patients are given little practical support to carry out the recommended actions, and health professionals' responses may render their attempts to act to secure their own safety ineffective. Some messages suggest an inappropriate shifting of responsibility onto patients. Advice that involves checking on or challenging health professionals' actions appears to be particularly problematic for patients. Such behaviors conflict with the expectations many people have--and think health professionals have--of patients' roles. DISCUSSION: A serious commitment to optimizing patients' contributions to safe care requires a research-based understanding of patients' perspectives and more practical facilitation of patient involvement. PMID- 16255327 TI - Critical laboratory value notification: a failure mode effects and criticality analysis. AB - BACKGROUND: The Failure Mode Effects and Criticality Analysis (FMECA) was applied to improve the timeliness of reporting and the timeliness of receipt by the responsible licensed caregiver of critical laboratory values (CLVs) for outpatients and non-critical care inpatients. METHODS: Through a risk prioritization process, the most important areas for improvement, including contacting the provider, assisting the provider in contacting the patient, and educating the provider in follow-up options available during off hours, were identified. ACTIONS TAKEN: A variety of systemic improvements were made; for example, the CLV notification process was centralized in the customer service center, with databases to help providers select options and make arrangements for follow-up care and an electronic abstract form to document the CLV notification process. Review of documentation and appropriateness of CLV follow-up care was integrated into the quality monitoring process to detect any variations or problems. RESULTS: The average CLV notification time for the month steadily declined during an eight-month period. Compliance was 100% for the "read-back" requirement and documentation in patient's health record. DISCUSSION: This proactive risk assessment project successfully modified the CLV notification program from a high- to a low-risk process, identified activities to further improve the process, and helped ensure compliance with a variety of requirements. PMID- 16255328 TI - Patient satisfaction with pain control during hospitalization. AB - BACKGROUND: The Joint Commission on Accreditation of Healthcare Organizations standards for pain management for hospitals took effect on January 1, 2001. A study was conductedto determine whether, and to what degree, patients' ratings of their satisfaction with pain control had changed during the four years after implementation of the Joint Commission standards compared with a baseline period of 2(1/2) years earlier. Patient satisfaction with pain control is a valid measure of a hospital's efforts to assess pain management and implementation of Joint Commission standards, insofar as it reflects the subjective nature of the pain experience. METHODS: Survey data representing identical hospitals for 26 quarters (2(1/2) years before and 4 years after January 1, 2001) were analyzed. More than 3,000,000 surveys, from 240 hospitals across the United States, were included in the analysis. RESULTS: The average score for all patients treated before the new standards were put into place was 85.2 (sigma = 19.7), while the average for those treated after that date was 85.7 (sigma = 19.6). Although small in absolute size, the difference is statistically significant at the .001 level. DISCUSSION: Although satisfaction with pain control varied within a relatively narrow range in the 10 quarters before the institution of the Joint Commission standards, it has subsequently shown an overall upward trend, as well as a cyclical pattern. Such evidence for even a modest increase might encourage caregivers to continue their efforts to monitor their patients' pain and to help patients control it. PMID- 16255329 TI - Medication dosing errors for patients with renal insufficiency in ambulatory care. AB - BACKGROUND: Approximately 8.7 million (3%) Americans have various degrees of chronic renal insufficiency (CRI). Patients with CRI are susceptible to adverse drug events related to improper dose adjustment of drugs that are eliminated primarily unchanged through the kidney. Renal dosing errors are an important quality-of-care problem in the inpatient setting, yet little is known about dosing errors for patients with renal insufficiency in the outpatient setting. METHODS: Electronic records were queried to identify patients with CRI (estimated creatinine clearance < 50 mL/min) who visited the ambulatory care clinic at least once from January 1, 2003 through December 31, 2003. RESULTS: Of the total of 224 patients identified with CRI, 157 (70%) received one or more of 17 drugs with high rates of renal elimination. A total of 207 drugs requiring dose adjustment were prescribed to these patients, and 52 (25%) were prescribed at an inappropriately high dose. For 127 (57%) of the 224 patients, CRI was not documented. Patients with documented CRI were equally likely to be prescribed an inappropriately high dose of a target drug. DISCUSSION: Incorrect dosing of medications among patients with CRI is common in the ambulatory care setting. Strategies for preventing medication dosing errors can target the prescribing and monitoring stages of pharmaceutical care. PMID- 16255330 TI - Preventing surgical fires: who needs to be educated? AB - BACKGROUND AND CASE STUDY: Surgical fires are rare but preventable. During facial surgery for a 68-year-old man, a fire broke out, resulting in first- and second degree burns after a nasal cannula ignited in an oxygen-rich environment because of improper draping and tenting. DISCUSSION: Operating room (OR) fires can be prevented if any component of the "fire triangle"-fuels, ignition sources, and oxidizers-is reduced or eliminated. The use of supplemental oxygen in the OR via nasal cannulae, nebulizers, and oxygen cylinders must always considered a potential source of fire. Deficits in knowledge among the surgical team with respect to the prevention and management of surgical fires were apparent. A plan was put into place to improve fire safety education, entailing an educational program that is included in intern and resident orientation. Surgical fire safety training was also put into place for anesthesia and surgical faculty. The anesthesia preoperative evaluation was modified to include an assessment of the patients' ability to tolerate short periods without oxygen. Posters and signs are now displayed in each OR suite. A complete policy review and update ensures that at least two fire drills are performed annually. CONCLUSION: Surgical fires can usually be prevented by educating staff about risk and prevention strategies. Such education should be part of all undergraduate medical, nursing, and other allied health profession education. PMID- 16255331 TI - Medication errors involving wrong administration technique. AB - Wrong administration technique has consistently been one of the most harmful types of medication error in health systems participating in MEDMARX. Administration technique errors typically are made by nurses administering medications on the patient care unit, although errors in administration technique also occur in other phases of medication use and involve other health care personnel and locations. The most commonly reported causes of error have been performance deficit, failure to follow procedures or protocols, and knowledge deficit. Educating and training health care personnel on proper administration techniques and use of infusion pumps and dispensing devices could reduce the risk of error. The drug products most often associated with administration technique errors and patient harm could be targeted in staff education and training programs. PMID- 16255332 TI - Using the AHRQ quality indicators to improve health care quality. AB - In summary, the AHRQ QIs are a set of readily available programs that can be downloaded without charge from the AHRQ Web site. The methodology is completely open and accessible to all users. The QI software can be applied to hospital administrative data that is available within individual institutions or from state data organizations and hospital associations and can provide valuable insights into health care quality at extremely low cost. The QIs have been incorporated into numerous quality assessment reports, including hospital specific reports, with the aim of improving health care quality at a reasonable cost. With enhancements currently underway, the QIs will be an even more valuable part of the toolkit to improve health care quality in the United States. PMID- 16255333 TI - A new profession in the pre-hospital care field--the ambulance nurse. PMID- 16255334 TI - Does intermediate care minimize relocation stress for patients leaving the ICU? AB - Relocation stress is a phenomenon in which physical and psychological disturbances are experienced following transfer from one environment to another [Carpenito LJ. (2000). Nursing Diagnosis. Application to Clinical Practice, 8th edn]. The purpose of this review was to identify whether a period of intermediate care minimizes the problems associated with relocation stress after discharge from the intensive care unit (ICU) and before transfer to the ward. Methods of retrieving the literature involved identifying key terms, utilizing a range of databases and applying specific criteria in order to delineate the boundaries of the search. Using electronic and manual search methods, 11 studies were selected, both primary and secondary research. Following tabulation and critiquing of the studies, the findings of the review suggest that the factors which contribute towards relocation stress are the loss of one-to-one nursing, a reduction of visible monitoring equipment, lack of continuity of care and inadequate preparation of the patient for the transfer. The evidence also indicates that in order to minimize these factors, early planning and preparation of the patient for transfer are required, incorporating strategies of gradual reduction in nursing attention and monitoring equipment and the provision of information. Although the benefits of intermediate care are established as being advanced monitoring, appropriate nurse-to-patient ratio, heightened demonstration of expert knowledge and skill, there is no sufficient evidence to indicate a period of intermediate care that can ease the transition from the ICU to the ward. PMID- 16255335 TI - Optimizing the fluid management of ventilated patients with suspected hypovolaemia. AB - Fluid management is a vital component of patient care within the critical care setting; it has a range of indications and applications, one of the most important being to maintain tissue perfusion and safeguard against single/multiple organ failure. Hypovolaemia is a commonly encountered condition within critical care and has the potential to jeopardize tissue perfusion and accelerate the risk of organ failure. In an attempt to optimize the fluid management of patients within the intensive therapy unit, this article outlines the development and implementation of a fluid prescription for ventilated patients with suspected hypovolaemia. PMID- 16255336 TI - What is supportive when an adult next-of-kin is in critical care? AB - There is little documented knowledge about what is supportive from the perspective of relatives with a critically ill next-of-kin in the intensive care unit (ICU). The aim of the present study was to generate a theoretical understanding of what relatives experience as supportive when faced with the situation of having an adult next-of-kin admitted to critical care. The study was designed using a grounded theory methodology. Interviews were conducted with 29 adult relatives of adult ICU patients in southwest Sweden. Relatives described the need to be empowered and that support was needed to enable them to use both internal and external resources to cope with having a next-of-kin in critical care. To achieve empowerment, the relatives described the need to trust in oneself, to encounter charity and to encounter professionalism. The findings can contribute understanding and sensitivity to the situation of the relatives as well as indicating what form social support should take. It is essential that healthcare professionals understand how important it is for relatives to have control over their vulnerable situation and that they also reflect upon how they would like to be treated themselves in a similar situation. Recommendations for future practice are presented. PMID- 16255337 TI - Mental Capacity Act 2005 England and Wales: a short summary. PMID- 16255338 TI - Iron ingestion: an evidence-based consensus guideline for out-of-hospital management. AB - From 1983 to 1991, iron caused over 30% of the deaths from accidental ingestion of drug products by children. An evidence-based expert consensus process was used to create this guideline. Relevant articles were abstracted by a trained physician researcher. The first draft of the guideline was created by the primary author. The entire panel discussed and refined the guideline before its distribution to secondary reviewers for comment. The panel then made changes in response to comments received. The objective of this guideline is to assist poison center personnel in the appropriate out-of-hospital triage and initial management of patients with suspected ingestions of iron by 1) describing the manner in which an ingestion of iron might be managed, 2) identifying the key decision elements in managing cases of iron ingestion, 3) providing clear and practical recommendations that reflect the current state of knowledge, and 4) identifying needs for research. This guideline applies to ingestion of iron alone and is based on an assessment of current scientific and clinical information. The expert consensus panel recognizes that specific patient care decisions may be at variance with this guideline and are the prerogative of the patient and the health professionals providing care, considering all of the circumstances involved. The panel's recommendations follow; the grade of recommendation is in parentheses. 1) Patients with stated or suspected self-harm or who are victims of malicious administration of an iron product should be referred to an acute care medical facility immediately. This activity should be guided by local poison center procedures. In general, this should occur regardless of the amount ingested (Grade D). 2) Pediatric or adult patients with a known ingestion of 40 mg/kg or greater of elemental iron in the form of adult ferrous salt formulations or who have severe or persistent symptoms related to iron ingestion should be referred to a healthcare facility for medical evaluation. Patients who have ingested less than 40 mg/kg of elemental iron and who are having mild symptoms can be observed at home. Mild symptoms such as vomiting and diarrhea occur frequently. These mild symptoms should not necessarily prompt referral to a healthcare facility. Patients with more serious symptoms, such as persistent vomiting and diarrhea, alterations in level of consciousness, hematemesis, and bloody diarrhea require referral. The same dose threshold should be used for pregnant women, however, when calculating the mg/kg dose ingested, the pre pregnancy weight of the woman should be used (Grade C). 3) Patients with ingestions of children's chewable vitamins plus iron should be observed at home with appropriate follow-up. The presence of diarrhea should not be the sole indicator for referral as these products are often sweetened with sorbitol. Children may need referral for the management of dehydration if vomiting or diarrhea is severe or prolonged (Grade C). 4) Patients with unintentional ingestions of carbonyl iron or polysaccharide-iron complex formulations should be observed at home with appropriate follow-up (Grade C). 5) Ipecac syrup, activated charcoal, cathartics, or oral complexing agents, such as bicarbonate or phosphate solutions, should not be used in the out-of-hospital management of iron ingestions (Grade C). 6) Asymptomatic patients are unlikely to develop symptoms if the interval between ingestion and the call to the poison center is greater than 6 hours. These patients should not need referral or prolonged observation. Depending on the specific circumstances, follow-up calls might be indicated (Grade C). PMID- 16255339 TI - Effect of high doses of sodium bicarbonate in acute organophosphorous pesticide poisoning. AB - The sodium bicarbonate (NaHCO3) dosage used in treating the patients with organophosphorous pesticide (OP) poisoning did not result in sufficient alkalinization of the blood. We thus investigated the effects of higher doses by infusion (5 mEq/kg over 1 hour, followed by 5-6 mEq/kg daily until recovery or death) to maintain the arterial pH between 7.45 and 7.55. There were 26 (14 M and 12 F) patients aged 24.1 +/- 9.7 years and 27 (16 M and 11 F) aged 25.7 +/- 9.1 years in the test and contro groups, respectively. Arterial blood pH of the test group (7.48 +/- 0.02) was much higher (p < 0.0001) than the controls (7.36 +/- 0.02). The total atropine doses used for the control patients (129.45 +/- 61 mg) was significantly (p = 0.048) higher than the controls (93.4 +/- 59.1 mg). Hospitalization days were statistically higher (p = 0.037) in the controls (5.59 +/- 1.97 days) than in the bicarbonate group (4.33 +/- 1.99 days). Infusion of high doses of NaHCO3 appears to be beneficial in treatment of patients with OP poisoning. PMID- 16255340 TI - Epidemiology of hydrogen phosphide exposures in humans reported to the poison center in Mainz, Germany, 1983-2003. AB - BACKGROUND: Poisonings with rodenticides containing hydrogen phosphide-releasing compounds may lead to deleterious organ dysfunction and death. Since data of hydrogen phosphide poisonings is limited to case reports/series, this study was intended to elucidate hydrogen phosphide poisonings based on a 20-year data collection. METHODS: Explorative data analysis of the Poison Center Mainz database looking for route of exposure, symptoms, and severity using the Poisoning Severity Score. RESULTS: From 1983-2003, 188 hydrogen phosphide poisonings were reported. Sixty-five percent of these were unintentional residential, 28% attempts to commit suicide (intentional), 5% occupational, and 2% undetermined. In the majority of intentional poisonings the poison was ingested, whereas in unintentional poisoning of adults inhalation exposure dominated, caused by inappropriate self-protection from the released hydrogen phosphide gas during usage. Frequently observed symptoms in unintentional poisonings were nausea, vomiting, pain, coughing, and dizziness with no further worsening of symptoms. In intentional poisonings frequent symptoms were vomiting, somnolence, seizures, coma, and shock with two initially fatal poisonings. Follow up on these cases showed a significant worsening of symptoms and a two-fold increase in fatal poisonings. CONCLUSION: Route of exposure, severity of symptoms, and the necessary treatment differs substantially between unintentional and intentional poisonings. In this study, two initially symptomatic intentional poisonings were later reported fatal. Careful monitoring is recommended in symptomatic intentional poisonings. PMID- 16255341 TI - Ajuga nipponensis Makino poisoning. AB - A 58-year-old man ingested an herbal preparation of Ajuga nipponesis Makino, as recommended in folk medicine for the treatment of hepatoma. He developed profound gastrointestinal upset immediately, and decreasing urine output and bilateral leg edema over the following 2 days. Notable laboratory findings included elevated levels of blood urea nitrogen, creatine, bilirubin, and hepatic transaminases. Deterioration of renal function was noted during hospitalization and he died 11 days after ingesting the herbal preparation. Two other healthy individuals also consumed the same herbal preparation at the same time but developed only vomiting and diarrhea. One or more of the four major components of Ajuga nipponesis Makino may be responsible for the renal toxicity found in our patient. PMID- 16255342 TI - Fatal unintentional overdose of diltiazem with antemortem and postmortem values. AB - BACKGROUND: Therapeutic errors involving calcium channel antagonists (CCA) resulting in death rarely have been reported in detail. We report a fatality from an unintentional overdose of sustained-release (SR) diltiazem including antemortem and postmortem blood concentrations. CASE REPORT: A 65-year-old man with aortic stenosis mistakenly took six tablets of diltiazem 360 mg SR. He developed symptoms of toxicity by 7 hours after ingestion. By 10 hours, he went to the emergency department. Despite a prolonged resuscitative attempt, the patient died 17 hours postingestion. An antemortem blood sample drawn 11.5 hours after ingestion was 2.9 mcg/mL. Postmortem gas chromatography of central blood revealed a diltiazem level of 6 mcg/mL and the peripheral blood sample measured 5 mcg/ mL. CONCLUSION: This case suggests that an unintentional overdose with a CCA may be lethal if the patient's cardiovascular ability to compensate for the toxic effects is compromised. PMID- 16255343 TI - A case of acute cholestatic hepatitis associated with the seeds of Psoralea corylifolia (Boh-Gol-Zhee). AB - The potential hepatotoxicity of herbal remedies is usually ignored in daily life. One such compound, Boh-Gol-Zhee (in Chinese, Bu Ku Zi), appeared to be associated with the occurrence of acute cholestatic hepatic injury in the following case. Some alternative medicine therapists claim that Psoralea corylifolia is effective for the treatment of osteoporosis. We observed a case of acute cholestatic hepatitis associated with the use of the seeds of Psoralea corylifolia in amounts over 10 times the usual dose in a postmenopausal woman. Liver biopsy showed zone three necroses, degenerating cells, cholestasis, and infiltrations with inflammatory cells. This case stresses the need to warn of the potential hepatotoxicity of the seed of Psoralea corylifolia, especially in a large dose. PMID- 16255345 TI - The future of HIGPA, part deux. PMID- 16255344 TI - Pacific volcanoes, mercury contaminated fish, and Polynesian taboos. PMID- 16255346 TI - More to bundling than meets the eye. Interview by Robert Neil. PMID- 16255347 TI - A survey of materials managers about their GPOs. From the buyer's perspective. PMID- 16255348 TI - Getting the skinny on lean management. PMID- 16255350 TI - Nurses benefit from supply chain overhaul. PMID- 16255349 TI - On the road to efficiency. PMID- 16255351 TI - When choosing medical devices, know their classification. PMID- 16255352 TI - Chlorhexidine-containing glass ionomer cement. A clinical investigation on the fissure caries inhibiting effect in first permanent molars. AB - Glass ionomer cement with addition of chlorhexidine used as a varnish on tooth surfaces has been shown to reduce the number of interproximal mutans streptococci (ms). The effect of a single application of such a varnish containing 2.5% chlorhexidine on occlusal caries development of the first permanent molars on 6 year-old children in a high caries incidence area was investigated. The children were examined according to WHO criteria and 262 children with 2 caries free contra-lateral molars were selected for treatment. Salivary ms samples were collected using the Strip-mutans (SM) method. After brushing the occlusal surfaces with a toothbrush and pumice in water slurry, rinsing and drying with a cotton roll, glass ionomer cement containing chlorhexidine (GI-CHX) and glass ionomer (GI) were applied randomly with a micro brush and the varnish was covered with occlusal wax. At baseline, the mean defs was 18.18 and DMFS was 0.25 and after one year 18.24 and 0.83 respectively. The salivary ms scores were high or very high (SM 2 and 3) in 85.6% at baseline. At the one-year follow up, the GI CHX and GI materials could not be detected in the fissures. Also, a large number of fissure sealants had been placed in the molars outside the study protocol; thus 4% of the GI-CHX and GI, and 70% of the untreated were sealed at year one. Overall, there was no significant difference between the caries-reducing effect of GI-CHX and GI, but a trend towards a higher effect was seen for GI-CHX. Excluding the sealed molars the reduction was 74% in the GI-CHX-group, and 71% in the GI-group. CONCLUSION: Addition of 2.5% chlorhexidine to glass ionomer did not seem to increase the caries-reducing effect of the varnish in this high caries incidence population. PMID- 16255353 TI - Patient flows in the care process of mandibular third molar surgery. AB - Our aim was to describe patient flows in mandibular third molar surgery at oral and maxillofacial specialist units. Our hypothesis was that there are variations in how care is delivered and that the variations could be explained by inter individual variations in surgeons' practice, the quality of the radiographs appended to the referral, and the staffing of the specialist units. A flow chart was constructed to simulate all possible patient flows in the care process. The chart begins with treatment planning, which was drawn up based on documents from the referring dentist or another caregiver; continues with the care process at the oral and maxillofacial surgery unit, including surgical consultations and radiological examinations; and ends with surgery. Surgeons at four oral and maxillofacial surgery units in the National Health Service in southern Sweden participated. The intention was to collect data on at least 100 patients who had undergone mandibular third molar surgery at each unit. Data on 361 patients were collected. The radiographs appended to the referral were judged to be inappropriate for the majority of the patients (61%). For 13% of these patients, supplementary radiographic examinations were made at the radiology clinic included in the unit, whilst 48% were examined at the oral and maxillofacial surgery clinic. There were eight different patient flow patterns. In one unit with three surgeons, eight different flow patterns were recorded, indicating an interindividual variation among the surgeons. In a second unit, six different flow patterns were recorded. In the last two units, the patient flows appeared to be the same at each unit, although the predominant patient flows in these two units differed. The number of patient visits to the specialist units ranged between one and three. In three specialist units, most patients were called twice whilst in one specialist unit most patients were called only once, to have the third molar removed. Differences existed in the care process. Overall, the number of patient visits seemed not to depend on whether the preoperative radiographic examination was judged to be appropriate or whether the additional radiographs were made at the radiology clinic. PMID- 16255354 TI - A Fourier transform Raman spectroscopy analysis of the degree of conversion of a universal hybrid resin composite cured with light-emitting diode curing units. AB - The degree of conversion (DC), of a universal hybrid resin composite cured with LED curing units with low and high power densities and a 510 mW/cm2 quartz tungsten halogen unit, was investigated with Fourier Transform Raman spectroscopy. Three curing depths (0, 2, 4mm) and 0 and 7 mm light guide tip - resin composite (LT - RC) distances were tested. The DC of the LED units varied between 52.3% - 59.8% at the top surface and 46.4% - 57.0% at 4 mm depth. The DC of specimen cured with a 0 mm LT- RC distance at 4 mm depth varied between 50.8% 57.0% and with 7 mm distance between 46.4% - 55.4%. The low power density LED unit showed a significantly lower DC for both distances at all depth levels compared to the other curing units (p < 0.05). Significant differences between the other curing units were only found at the 4 mm depth level cured from 7 mm distance (p < 0.05). The reduction in DC by increasing LT- RC distance was less than 10% for all curing units. It can be concluded that the improved LED curing units could cure the studied resin composite to the same DC as the control unit. PMID- 16255355 TI - A Swedish version of the GOHAI index. Psychometric properties and validation. AB - This study investigated a Swedish version of the 12 item General Oral Health Assessment Index (GOHAI). The aim was to validate the translated instrument, and to investigate factors that may influence the GOHAI score. Consecutive samples at eight dental clinics in Goteborg, Sweden were asked to answer the GOHAI, the short form of Oral Health Impact Profile (OHIP-14) and a questionnaire including socio-demographic, general health and oral health questions. 153 patients (50-89 yrs old) out of 237 (65%) returned the questionnaires. The most commonly reported problem (GOHAI) was 'worried or concerned' due to problems related to oral health (48%), followed by 'unhappy with the appearance of teeth, gums, or dentures' (36%) and 'teeth sensitive to hot, cold, or sweets' (33%). The Cronbach's Alpha (0.86) indicated a high degree of internal consistency and homogeneity between items. Item-scale correlations varied between 0.50 and 0.83. Strong relations were found between depressed GOHAI-scores and dental problems (mobile teeth, number of teeth, dentures). The correlation between the GOHAI score and the OHIP 14 was high (-0.83) indicating good construct validity. Also 'perceived general health' and 'satisfaction with life-situation' and dental status were significantly, but less strongly, correlated with the patient's GOHAI score. In a linear regression analysis, age,'perceived general health' and dental status (number of teeth in lower jaw and mobile teeth) were found to have a significant effect on the GOHAI score. Factor analysis of GOHAI indicated a two-factor solution and did not support the theoretical construction reported of the index. The test-re-test reliability was assessed in a separate sample (members of a physical handicap organization, n = 47) and the correlation coefficient for the GOHAI was 0.64. For individual items, the weighted kappa coefficient varied between 0.25 and 0.80. In conclusion, the Swedish version of the GOHAI showed acceptable reliability and validity. PMID- 16255356 TI - [Satanic ritualistic murders and child abuse--evidence-basing versus ideology]. PMID- 16255357 TI - [What is a detailed medication record? It contains indication, preparation, concentration, dosage--as well as an administration schedule]. PMID- 16255358 TI - [Old gender patterns still used when physicians are pictured in medical journals. Male physicians are portrayed as active leaders, women physicians as listening, compassionate]. AB - As a part of a medical student' s research project on medical socialisation, the 2002 issue of two Swedish medicaljournals, Lakartidningen and Moderna Lakare, were scrutinized regarding how male and female physicians were represented on pictures. The outward facade was mostly male; 87% of portrays of editorials pictured a man, 81% of career announcements displayed men. Authors of articles and chroniclers more often presented a female face (55%). Photos in reports showed around 60% men and 40 % women, both regarding area and number of photos. These shares corresponded well to the actual share of male and female physicians in Sweden. The content analysis of pictures, however, demonstrated gender features: men were to a much higher degree focused in leading, demonstrating and speaker positions, while women to a higher degree were portrayed as taking part in consultations or caring activities. PMID- 16255360 TI - ["I'm just a civil servant--neutral and sexless". About the resistance against the gender perspective and the risk of gender bias in medicine]. AB - In medicine a gender perspective includes taking into consideration not only biology but also the cultural and psychosocial aspects of being a woman or a man. Unawareness or denial of gendered norms in society and/or of the power asymmetry between women and men can lead to resistance to gender issues and to the risk of gender bias in clinical medicine as well as in medical research and education. In a theoretical model presented in this paper the resistance and the bias risk are analysed in relation to assumptions about sameness/difference and equity/inequity between women and men. Such an analysis could facilitate the implementation of a gender perspective in settings and situations where resistance to gender issues is met. PMID- 16255359 TI - [The laundry-basket project--gender differences to the very skin. Different treatment of some common skin diseases in men and women]. AB - In this study, we have analysed the treatment traditions in a dermatological outpatient clinic, from a gender perspective. Eczema and psoriasis were of similar frequency in male and female patients, while a higher number of female patients had eczema of the hands. Ultraviolet light (UV) treatment was given to a higher number of male patients in all diagnostic groups including eczema of the hands. In addition, a higher number of given treatments was given to the male patients than to the female patients. As for prescriptions to male and female patients from our department and in Stockholm county (1.8 mill.), a much larger amount of preparations for local treatment was received by female patients, especially emollients. Male patients had received more of calcipotriol creme in addition to their being treated more intensively at the clinic. In a sub-group analysis on patients with psoriasis vulgaris on our clinic and in a patient cooperation-based treatment department, we found the same relation between male and female treatment as in the larger group. On follow-up, the number of female patients with psoriasis who were given treatment at our clinic had increased, but the number of treatments given to men was still higher than for female patients. An economic analysis of these findings show a great overweight of clinic-based treatment costs for male patients, while female patients receive emollients for self-care to a greater extent. We conclude that more studies are needed to clarify the basis of these findings. PMID- 16255361 TI - [Should obesity in the elderly be treated? Sparse scientific evidence to support treatment of overweight in this age group]. AB - Mean body weight increases with age up to about ago 60 and then levels off. Information about the association between body weight and mortality at old age is sparse, since most studies are cross-sectional. Some studies even suggest a protective effect of overweight at old age. Weight loss treatment may be indicated even at old age to relieve mechanical problems, reduce the need for pharmacotherapy and improve self-esteem. As regards treatment, most clinical drug trials actively exclude older people, and hence little is known about the effects in this age group. However, physical activity, even at old age, has documented beneficial effects, in spite of modest effects on weight. Bariatric surgery, although obviously debatable in this age group, has been safely performed. With an increased segment of the population of higher age under way, strategies to manage obesity, also in this age group, need to be developed. PMID- 16255362 TI - [Step by step approach--a method of working in stroke rehabilitation]. AB - Modern stroke rehabilitation should be conducted on a step by step approach towards final achievable objectives. Guidelines about how to introduce this in daily work are however missing. The aim of the present study was to create and introduce a paradigm with intermediate and final objectives in a multidisciplinary stroke team. METHOD: Over a period of eight months the staff was trained to use team conferences to evaluate and formulate intermediate and final objectives. The staff s ability to formulate correct objectives and their knowledge about previously agreed targets were evaluated monthly through a questionnaire. RESULTS: The number of objectives that were accurately formulated improved from 50% to 90% and the caring team' s awareness of these improved from 45% to >80g. CONCLUSIONS: This model can be used to improve the multidisciplinary caring team's ability to formulate objectives and keep current objectives in mind in the daily work in a stroke unit. PMID- 16255363 TI - [Contrast-induced nephropathy after computer tomography. Hydration and adapted contrast media dosage for the best prophylaxis]. PMID- 16255364 TI - [Medication records reduce medication errors when transferring and discharging hospitalized patients]. PMID- 16255365 TI - ["There is nothing in life to be afraid of, all can be understood--now is the time to understand more, so as to fear less"]. PMID- 16255366 TI - [Diffuse terms with certain diffusion]. PMID- 16255367 TI - [Two comments (The Green Party) on complementary medicine: we do not ignore the research, but we want to emphasize the new issues]. PMID- 16255368 TI - [We want increased cooperation between complementary medicine and medicine]. PMID- 16255369 TI - [A textbook with misleading information about whiplash injuries]. PMID- 16255370 TI - [The Medical Products Agency didn't "choose" Artrox]. PMID- 16255371 TI - [We think, thus we have a soul]. PMID- 16255372 TI - [It's not too late to take sides with the patients!]. PMID- 16255373 TI - [Fantastic results of reconstructive orthopedic surgery]. PMID- 16255374 TI - [Cholesterol and Alzheimer's disease]. AB - Epidemiological, biochemical and pharmacological investigations provide increasing number of evidences that altered cholesterol metabolism contributes to the development of Alzheimer's disease. The objective of the present paper is to review existing information about the links of cholesterol and amyloid metabolism from the clinical and proposed etiological viewpoints of the most frequent dementing disorder in Hungary. Beta-amyloid peptide, the major component of the senile plaques in the Alzheimer's disease brains is the end product of the abnormal post-translational processing of its precursor, called amyloid precursor protein. The compartmentation of the amyloid precursor protein molecule within the cell membrane is regulated by the cholesterol content of the bilayers. The amyloid precursor protein molecule could be present either in-, or outside of the membrane rafts. Any kind of process, which alters the compartmentation preference of the amyloid precursor protein molecule, by transferring it to the membrane rafts, favours beta- and gamma-secretase cleavage, and should be recognised as an amyloidogenic process. If the blood-brain barrier is intact, the brain is not able to take up the lipoprotein particles responsible for the transport of cholesterol. Instead of the active uptake, neurons and glial cells synthetize cholesterol de novo, in a process, where the rate limiting enzyme is 3-hydroxy-3 methylglutaryl coenzyme A. On the other hand, the brain specific CYP46A1 enzyme is responsible for the degradation of cholesterol into a water soluble metabolite, called 24S-OH cholesterol. The decreased CYP46A1 activity in the brain of Alzheimer's disease patients raises membrane cholesterol levels, and as a consequence the amyloid precursor protein is shifted and deposited in the cholesterol rich lipid rafts leading to beta-amyloid peptide specific metabolism. Among the polymorphic variants of the apolipoprotein E gene, the E4 allele is considered as a major risk factor for Alzheimer's disease. The E4 allele carrier Alzheimer's disease probands have increased amyloid burden, decreased beta amyloid peptide degradation, and less effective neuronal repair mechanisms. Even as early as age 30, patients with Niemann-Pick Type C disease show clinical and neuropathological signs of Alzheimer's disease. The point mutation of the protein responsible for the endosomal transport of cholesterol is considered as a major cause of the beta-amyloid peptide deposition in the brain of Niemann-Pick Type C patients. One of the most exciting recent discovery, that Niemann-Pick Type C disease could be recognised as a disease model for Alzheimer's disease. New, promising cholesterol metabolism related therapeutic approaches are discussed, but it is emphasized that the clinical evidences regarding their efficacy in Alzheimer's disease are still missing. PMID- 16255375 TI - [Microscopic colitis]. AB - AIM: Various case presentations in the literature discussed that frequent watery diarrheas introduce the clinical picture of microscopic colitis and intermittent or continuous diarrhea can remain. On the other hand, numerous cases never suffered from episodes of diarrhea but suffered from chronic constipation definitely. The authors present their first observations in Hungary. METHODS: A cohort of patients with independently confirmed typical histopathological changes was investigated. 53 patients with histologically proved MC (46 with CC, 7 with LC) were included. The existence of diarrhea or constipation and the co-existence of autoimmune diseases also investigated and all data were retrospectively analyzed. RESULTS: In contrast of the literature, about half of the patients had chronic constipation and cca. 50% of MC patients had autoimmune disease and the diagnosis of autoimmune disease was always prior to MC. CONCLUSIONS: Causes of inducing and amplifying the processes of "remodelling" the gut wall are thought to be unknown, but the results are: altered structure (and function?). All useful observations of this study may enrich the practice and can help these patients. PMID- 16255376 TI - [Experiences with serum BNP (B type natriuretic peptide) in patients with systolic and diastolic heart failure]. AB - Current study confirms and extends recent observations concerning the diagnostic usefulness of B type natriuretic peptide. It discriminates well between dyspnea of cardiac and non-cardiac origin in the unselected population. Echocardiography represents useful diagnostic tool for assessment of systolic and diastolic ventricular function. In diastolic heart failure the elasticity of left chamber decreases due to the increase of the filling pressure. B type natriuretic peptide predominantly derived from the atrial tissue in patients with chronic heart failure. In the literature contradictory data has been found about serum B type natriuretic peptide level and diastolic dysfunction in cases with good left ventricular function. Authors reviewed 35-34 unselected patients with chronic systolic and isolated diastolic dysfunction. The serum B type natriuretic peptide level increased significantly in all the systolic heart failure patients, while, in patients with isolated diastolic heart failure the values were increased only cases with increased atrial value (calculated to the body surface). PMID- 16255377 TI - [Fermented wheat germ extract in the supportive therapy of colorectal cancer]. AB - The role of the product in the treatment of colorectal cancer is reviewed in the light of experimental and clinical results to date. The fermented wheat germ extract (code name: MSC, trade name: Avemar) registered as a dietary food for special medical purposes for cancer patients to complement the active oncotherapy, exerted a growth inhibitory effect in HCR-25 human colon carcinoma xenograft, and had a synergistic effect with 5-FU in mouse C-38 colorectal carcinoma. The product is capable of chemoprevention of colon carcinoma in F-344 rats. One of the most significant underlying mechanism is a highly cancer cell specific induction of caspase-3 mediated cleavage of PARP. In the frame of supportive therapy, fermented wheat germ extract proved to be efficient in the treatment of colorectal cancer in humans. 30 patients following radical operation were treated with standard postoperative therapy, 12 of them were given fermented wheat germ extract as additive treatment: following a 9 month long administration, no new distant metastases were detected, in contrast to 4 out 18 treated with standard therapy alone. Out of 34 patients following radical surgery and treated with chemotherapy, 17 who were given fermented wheat germ extract, achieved an improved survival rate. In the frame of a controlled multicenter open label cohort study, 170 colorectal cancer patients received anticancer therapies (chemo/radiotherapy) completed with fermented wheat germ extract in 66 of them. Results (fermented wheat germ extract vs. control): new recurrences: 3.0% vs. 17.3% (p < 0.01); new metastases: 7.6% vs. 23.1% (p < 0.01); deaths: 12.1% vs. 31.7% (p < 0.01), progression-related events in total: 16.7% vs. 42.3% (p < 0.001). Survival analysis showed significant improvements in the fermented wheat germ extract group, regarding progression-free (p = 0.0184) and overall survival probabilities (p = 0.0278). Strong predictors of survival determined by Cox's proportional hazards were UICC stage and fermented wheat germ extract treatment. Mild gastrointestinal side effects were observed in 9 cases. Supportive application of fermented wheat germ extract in colorectal cancer is highly recommended. PMID- 16255378 TI - [The first genetically supported case of chronic benign pemphigus (Hailey-Hailey disease in Hungary]. AB - Hailey-Hailey disease, or chronic benign pemphigus (MIM# 169600), is a genodermatosis arising in adult age with recurrent vesicles and erosions primarily in the flexural areas. It is an autosomal dominant skin disorder characterized by abnormal keratinocyte adhesion in the suprabasal layers of the epidermis. ATP2C1, encoding the human secretory pathway Ca(2+)-ATPase (hSPCA1), was recently identified as the defective gene in Hailey-Hailey disease. More than 82 different ATP2C1 mutations have been described up to date. In this study, a case of Hailey-Hailey disease is presented where a nucleotide change (1402C > T) in the decoding region of ATP2C1 resulted in a premature stop mutation (R468X). This defect has been reported earlier in a patient of European descent. A brief molecular genetic review of the disorder is also given. PMID- 16255379 TI - [Triplex antithrombotic therapy after coronary-stent implantation (that is absent from recommendation)]. PMID- 16255380 TI - Is the DeVries-Rose to Weber transition empirically possible with sine-wave gratings? AB - Visual functioning at various retinal illuminance levels is usually measured either by determining grating acuity as a function of light level or by determining how sensitivity to sine-wave gratings changes with retinal illuminance. The former line of research has shown that grating acuity follows a two-branch relationship with retinal illuminance, with the point of discontinuity occurring at the transition from scotopic to photopic vision. Results of the latter line of research have summarily been described as a transition from the DeVries-Rose law to Weber's law, according to which log sensitivity increases linearly with log illuminance with a slope of 0.5 over a range of low illuminances (the DeVries-Rose range) and then levels off and does not increase with further increases of illuminance (the Weber range). This paper aims at determining the compatibility of the results of these two lines of research. We consider empirical constraints from data bearing on the shape of the surface describing contrast sensitivity to sine-wave gratings as a function of spatial frequency and illuminance simultaneously, in order to determine whether they are consistent with a summary description in terms of DeVries-Rose and Weber's laws. Our analysis indicates that, with sine-wave gratings, the DeVries-Rose law can only hold empirically at low spatial frequencies. PMID- 16255381 TI - Multidimensional scaling of schematically represented faces based on dissimilarity estimates and evoked potentials of differences amplitudes. AB - This study researches the input of the cerebral occipital and temporal cortex in the analysis of facial configuration and expressive characteristics. Analysis is based on the construction of a spherical model for the differentiation of schematically presented faces with quantitatively altering curvature of the mouth and brows. The model is designed using the method of multidimensional scaling of the dissimilarity judgments between stimuli (faces) and the amplitude of evoked potentials of differences (EPD) between abrupt stimulus changes recorded from the occipital and posterior temporal cortex. Analysis of the structure of the spherical model of facial differentiation depending on the electrode site and the latency of the EPD component within the duration of 120-240 ms has demonstrated that the activity of the occipital and posterior temporal cortex of the right hemisphere is associated with the emotional characteristics of the presented face, whereas facial configuration is reflected in the activation of both posterior temporal cortex and the occipital cortex of the left hemisphere. At all electrode sites maximum information of the emotional expression and configuration is represented in inter-peak amplitude P120-N180. With increasing latency there is increased distortion of the structure of differences in the spherical model of schematically presented faces, which is interpreted as an attenuation of electrical activity associated with the analysis of the emotional expression, which occurs more rapidly than configuration analysis. PMID- 16255382 TI - Simulation of the filtering role of habituation to stimuli. AB - In the context of a medium-term study designed to integrate the simulation of different types and processes of learning-such as classical, operant, and some cognitive types--one must start with other more elementary ones that are facilitators of the more complex types and processes. Of special interest is habituation, owing to the filtering out of irrelevant stimuli, which means that the simulated agent does not have to respond to them. This paper presents two difference functions constructed to computationally simulate the characteristics that define habituation. The behavior of these functions is described, as are differences arising from stimulus intensity and interstimulus intervals. Results are compared with existing empirical data. PMID- 16255383 TI - A new mathematical model for assessment of memorization dynamics. AB - A new memory model is proposed based on regression analysis and exponential- shaped learning curves. The efficacy of the model is tested with several types of experiments including food aversion in snails, maze learning in rats and memory tests for adults and children. The model is also tested on drug abusers and alcoholics. The results of goodness of fit tests indicate that our model can accurately be used to predict the memory dynamics of diverse experiments and populations. The model can also be used to predict both group and individual performance. The application of the model to detect memory impairment is discussed, as are limitations. PMID- 16255384 TI - Lexical processing of ambiguous words: dominance or associative strength? AB - Four experiments examined the role of meaning frequency (dominance) and associative strength (measured by associative norms) in the processing of ambiguous words in isolation. Participants made lexical decisions to targets words that were associates of the more frequent (dominant) or less frequent (subordinate) meaning of a homograph prime. The first two experiments investigated the role of associative strength at long SOAs (Stimulus Onset Asynchrony) (750 ms.), showing that meaning is facilitated by the targets' associative strength and not by their dominance. The last two experiments traced the role associative strength at short SOAs (250 ms), showing that the manipulation of the associative strength has no effect in the semantic priming. The conclusions are: on the one hand, semantic priming for homographs is due to associative strength manipulations at long SOAs. On the other hand, the manipulation of the associative strength has no effect when automatic processes (short SOAs) are engaged for homographs. PMID- 16255385 TI - Constructivism, the so-called semantic learning theories, and situated cognition versus the psychological learning theories. AB - In this paper, the perspective of situated cognition, which gave rise both to the pragmatic theories and the so-called semantic theories of learning and has probably become the most representative standpoint of constructivism, is examined. We consider the claim of situated cognition to provide alternative explanations of the learning phenomenon to those of psychology and, especially, to those of the symbolic perspective, currently predominant in cognitive psychology. The level of analysis of situated cognition (i.e., global interactive systems) is considered an inappropriate approach to the problem of learning. From our analysis, it is concluded that the pragmatic theories and the so-called semantic theories of learning which originated in situated cognition can hardly be considered alternatives to the psychological learning theories, and they are unlikely to add anything of interest to the learning theory or to contribute to the improvement of our knowledge about the learning phenomenon. PMID- 16255386 TI - How impulsivity is related to intelligence and academic achievement. AB - This study investigated the relationships between impulsivity, intelligence, and academic failure in a sample of 241 secondary school students who completed Thurstone's (1938) Primary Mental Abilities (PMA) test and Dickman's (1990) and Barratt's (1985) impulsivity questionnaires (DII and BIS-10, respectively). Results show an inverse relationship between impulsivity and intelligence, specific to the scales with higher loadings on crystallized intelligence, and a positive relationship between impulsivity and academic failure. These results indicate that impulsivity is not directly related to intelligence and may act as a moderator variable between individuals' resources and their achievements. PMID- 16255387 TI - The mothering of conduct problem and normal children in Spain and the USA: authoritarian and permissive asynchrony. AB - Ninety-two clinic-referred and nonclinical mother-child dyads in Spain and the USA were observed in their home settings under naturalistic conditions for a total of 477 hours. Children in the clinic-referred dyads were considered troubled because of conduct problems. The observations were aimed at assessing two forms of mother-child asynchrony, either of which was expected to differentiate clinic referred from nonclinical dyads. Authoritarian asynchrony was defined as a mother's indiscriminate use of aversive reactions to her child, whereas the permissive form entailed indiscriminate positive reactions. Results showed the American mothers to generate more permissive asynchrony, whereas the Spanish mothers were inclined in the authoritarian direction. Only authoritarian asynchrony differentiated the clinical versus nonclinical dyads in each country. Discussion was centered on the greater salience of aversive as opposed to positive maternal attention, and cultural differences between countries that might have accounted for the different parenting styles. PMID- 16255388 TI - Does quantity generate quality? Testing the fundamental principle of brainstorming. AB - The purpose of this work is to test the chief principle of brainstorming, formulated as "quantity generates quality." The study is included within a broad program whose goal is to detect the strong and weak points of creative techniques. In a sample of 69 groups, containing between 3 and 8 members, the concurrence of two commonly accepted criteria was established as a quality rule: originality and utility or value. The results fully support the quantity-quality relation (r = .893): the more ideas produced to solve a problem, the better quality of the ideas. The importance of this finding, which supports Osborn's theory, is discussed, and the use of brainstorming is recommended to solve the many open problems faced by our society. PMID- 16255389 TI - Some variables associated with psychologists' appraisal of psychotherapy in Argentina. AB - The purpose of this research is to provide information about the state of psychotherapy in Argentina, as well as to study some variables involved in psychologists' evaluation of psychotherapy. A survey of 14 items was elaborated ad-hoc for this purpose, and administered to 226 psychologists from all over the country by mail and, in some cases, personally. Results suggest that the predisposition to apply techniques from different frameworks is associated with the amount of experience and with the perception of a bias concerning other theoretical focuses, although it is not related to the evaluation of the state of psychotherapy. On the other hand, in the view of psychologists, personal aspects of the therapeutic relationship are believed to be the most important factor in patients' change and effective progress. PMID- 16255390 TI - Evaluation of a group cue-exposure treatment for opiate addicts. AB - Twenty-four detoxified opiate addicts were randomized to an experimental group and a control group to evaluate efficacy of a group cue-exposure treatment to reduce or extinguish classically conditioned responses to drug-related stimuli. Assessment included psychophysiological responses (skin temperature, skin conductance level--SCL--, and heart rate) to a videotape and subjective measures (subjective craving, positive and negative affect) before and after the videotape. The experimental group received a group cue-exposure program to drug related stimuli that comprised twelve treatment sessions administered three times weekly. The treatment program significantly reduced conditioned responses to drug related stimuli, as measured by SCL and positive affect. PMID- 16255391 TI - Structure and stimulus familiarity: a study of memory in chess-players with functional magnetic resonance imaging. AB - A grandmaster and an international chess master were compared with a group of novices in a memory task with chess and non-chess stimuli, varying the structure and familiarity of the stimuli, while functional magnetic resonance images were acquired. The pattern of brain activity in the masters was different from that of the novices. Masters showed no differences in brain activity when different degrees of structure and familiarity where compared; however, novices did show differences in brain activity in such contrasts. The most important differences were found in the contrast of stimulus familiarity with chess positions. In this contrast, there was an extended brain activity in bilateral frontal areas such as the anterior cingulate and the superior, middle, and inferior frontal gyri; furthermore, posterior areas, such as posterior cingulate and cerebellum, showed great bilateral activation. These results strengthen the hypothesis that when performing a domain-specific task, experts activate different brain systems from that of novices. The use of the experts-versus-novices paradigm in brain imaging contributes towards the search for brain systems involved in cognitive processes. PMID- 16255392 TI - Neuropsychological disorders indicative of postresuscitation encephalopaty in rats. AB - The aim of this research was to study the effect of 12-minute clinical death on innate and acquired behavior, biogenic amine concentration, and the composition and quantity of neural populations in specific brain regions of white rats. The study shows that in animals during the postresuscitation period with formal restoration of neurological status, there are changes in emotional reactivity, orientation-exploration reactions, impairment of learning and memory, decrease in exercise tolerance and pain sensitivity. These processes are accompanied by alterations in serotonin and norepinephrine levels in the frontal cerebral cortex, dopamine and serotonin levels in the striatum, certain biochemical indices in blood plasma and neural loss in the CA1 sector of the hippocampus and lateral portions of the cerebellum. PMID- 16255393 TI - Sampling plans for fitting the psychometric function. AB - Research on estimation of a psychometric function psi has usually focused on comparing alternative algorithms to apply to the data, rarely addressing how best to gather the data themselves (i.e., what sampling plan best deploys the affordable number of trials). Simulation methods were used here to assess the performance of several sampling plans in yes-no and forced-choice tasks, including the QUEST method and several variants of up-down staircases and of the method of constant stimuli (MOCS). We also assessed the efficacy of four parameter estimation methods. Performance comparisons were based on analyses of usability (i.e., the percentage of times that a plan yields usable data for the estimation of all the parameters of psi) and of the resultant distributions of parameter estimates. Maximum likelihood turned out to be the best parameter estimation method. As for sampling plans, QUEST never exceeded 80% usability even when 1000 trials were administered and rendered accurate estimates of threshold but misestimated the remaining parameters. MOCS and up-down staircases yielded similar and acceptable usability (above 95% with 400-500 trials) and, although neither type of plan allowed estimating all parameters with optimal precision, each type appeared well suited to estimating a distinct subset of parameters. An analysis of the causes of this differential suitability allowed designing alternative sampling plans (all based on up-down staircases) for yes-no and forced-choice tasks. These alternative plans rendered near optimal distributions of estimates for all parameters. The results just described apply when the fitted psi has the same mathematical form as the actual psi generating the data; in case of form mismatch, all parameters except threshold were generally misestimated but the relative performance of all the sampling plans remained identical. Detailed practical recommendations are given. PMID- 16255394 TI - Inhaled corticosteroids in COPD. PMID- 16255395 TI - Experience with inhaled nitric oxide therapy in hypoxic respiratory failure of the newborn. AB - BACKGROUND: Respiratory diseases are the commonest cause of morbidity and mortality in newborns. Inhaled nitric oxide (iNO) has been shown to be effective in the management of persistent pulmonory hypertension of newborn (PPHN). OBJECTIVES: To retrospectively analyse data to determine the effectiveness of inhaled nitric oxide (iNO) in the management of newborns with PPHN in terms of survival and changes in oxygenation status. METHODS: Neo-natal data since inception of iNO therapy at the unit (past six years) was reviewed. Pertinent demographic and clinical information was collected from medical records of newborns that received inhaled nitric oxide therapy during their stay. Details of underlying illnesses, other therapeutic modalities, arterial blood gas, ventilatory and nitric oxide parameters were assessed and analysed to ascertain efficacy of iNO. RESULTS: A total of 36 babies (gestational age ranging from 24 41 weeks) received iNO during this period; two were excluded from final analysis. Overall survival rate was 80 percent. There was a statistically significant increase in systemic oxygenation (PaO2) from 41.1 +/- 2.1 mmHg to 128.5 +/- 13.2 mmHg and a decline in oxygenation index (OI) from 49.4 +/- 5.9 to 17.3 +/- 2.5, when assessed after four hours (P < 0.001). Mean duration of iNO therapy was 63 +/- 7.3 hours and the maximum methaemoglobin levels were noted to be 2.1 percent. CONCLUSIONS: Inhaled nitric oxide appears to be an effective rescue therapy for the management of PPHN associated with hypoxic respiratory failure. It is safe and well tolerated with no evidence of clinical or biochemical side effects. PMID- 16255396 TI - Pulmonary functions in Parkinson's disease. AB - BACKGROUND: Parkinson's disease (PD) is a common disabling and progressive neuro degenerative disorder. Morbidity and mortality are frequently associated with pulmonary dysfunction in patients with PD. AIM: To investigate characteristics of pulmonary function test (PFT) abnormalities in patients with PD and to study the effect of levodopa on the PFT findings. METHODS: Pulmonary function testing was performed in 35 patients with PD during 'off' and 'on' state while on levodopa; and 35 matched healthy controls. RESULTS: The forced vital capacity (FVC) (P < 0.001), maximum voluntary ventilation (MVV) (p < 0.001), maximum expiratory pressure (MEP) (p < 0.001), and maximum inspiratory pressure (MIP) (p < 0.001) were significantly reduced in patients with PD compared to controls. There was significant improvement in the PFT values in 'on' state compared to 'off' state. CONCLUSIONS: A restrictive pattern of pulmonary dysfunction is present in patients with PD, which improves significantly with levodopa. The evaluation and rehabilitation of respiratory disturbances should be routinely included in the management of patients with Parkinson's disease. PMID- 16255397 TI - Diagnosis of Pneumocystis pneumonia by bronchoalveolar lavage cytology: experience at a tertiary care centre in India. AB - BACKGROUND: Diagnosis of Pneumocystis pneumonia requires morphological demostration of P. carinii (now re-named as P. jiroveci). Although bronchoalveolar lavage (BAL) fluid cytology constitutes a formidable tool for detecting this infection, few studies on the utility of BAL cytology in diagnosing PCP are available from India. The present study reports the clinical spectrum, cytomorphological features and the utility of BAL cytology in diagnosing Pneumocystis infection from a tertiary care centre in India. METHODS: Retrospective study of 13 patients with PCP, diagnosed on examination of BAL fluid. RESULTS: The mean age of the patients was 41.2 years. One patient had human immunodeficiency virus (HIV) infection, while the other 10 were renal transplant receipients on immunosuppressive therapy. The immune status of two patients was unknown. Fever, cough and shortness of breath were the main presenting symptoms. Radiological diagnosis of Pneumocystis pneumonia was offered in only one case. Foamy alveolar casts were present in all cases. Silver methanamine stain enhanced the rounded, helmet or cleft forms of sporozoites. Inflammatory infiltrate was mainly polymorphonuclear. CONCLUSIONS: BAL cytology, thus, constitutes a useful diagnostic modality for morphological documentation and reliable diagnosis of Pneumocystis pneumonia in an immunocompromised host. Pneumocystis pneumonia appears to be a common opportunistic infection in renal transplant receipients in India. PMID- 16255398 TI - Noninvasive estimation of clinically asymptomatic pulmonary hypertension in idiopathic pulmonary fibrosis. AB - OBJECTIVES: To determine the prevalence of pulmonary hypertension in patients with idiopathic pulmonary fibrosis (IPF) by noninvasive methods. PATIENTS AND METHODS: Twenty seven patients with IPF attending the Chest Clinic over a period of one-and-a-half-years underwent echocardiography for evidence of pulmonary hypertension, which was defined as pulmonary artery systolic pressure > or = 40 mmHg by Doppler echocardiography, or pulmonary acceleration time < or = 100 milliseconds or two-dimensional echocardiographic findings of right ventricular hypertrophy or overload. RESULTS: Two patients with clinical evidence of pulmonary hypertension were excluded from analysis. Their mean age (n = 25) was 53.8 years. The mean duration of symptoms before presentation was 2.1 years. Pulmonary hypertension was diagnosed by echocardiography in nine of the 25 patients (36%). There was a statistically significant difference between the duration of illness, arterial oxygen tension (PaO2) levels and forced vital capacity in patients with pulmonary hypertension than in those without pulmonary hypertension. Logistic regression analysis showed a significant association between the duration of illness, decreasing forced vital capacity and hypoxemia with the development of pulmonary hypertension. CONCLUSIONS: Echocardiography detects pulmonary hypertension in clinically asymptomatic individuals, and should be used routinely for the diagnosis of pulmonary hypertension in patients with IPF. PMID- 16255399 TI - Human pneumocystosis. AB - Pneumocystis is an atypical fungus causing pneumonia in immuno-compromised individuals. Though previously termed as Pneumocystis carinii, the recent taxonomy has considered human derived Pneumocystis to be a different species Pneumocystis jiroveci. The organism is the most common cause of opportunistic infections among patients with acquired immunodeficiency syndrome (AIDS) in developed countries. Incidence of Pneumocystis pneumonia or pneumocystosis in developing countries including India continues to be low. Microscopy of appropriate clinical samples has been the mainstay of diagnosis of pneumocystosis. Amplification techniques are now being evaluated for detection of P. jiroveci. This review attempts to give a recent update on P. jiroveci with special focus on epidemiology, taxonomy, current diagnostic modalities and recommended immuno-prophylaxis. PMID- 16255400 TI - Gastric volvulus caused by paraesophageal hernia complicating mid-trimester pregnancy. AB - A 36-year-old woman, who was 19 weeks pregnant presented with epigastric pain and a one-week history of repeated vomiting. Endoscopy revealed twisted stomach. CT scan of the chest showed figure of eight stomach consistent with gastric volvulus. Confirmation of diagnosis was made by laparatomy in which reduction of the oedematous stomach and excision of ischemic omental patch and repair of a huge paraesophageal hernia were performed. Two days after operation, abortion took place. Few days later, rapid deterioration in renal and hepatic function occurred followed by maternal death. PMID- 16255401 TI - Pulmonary veno-occlusive disease in a patient with a history of Hashimoto's thyroiditis. AB - Pulmonary veno-occlusive disease (PVOD), a rapidly progressive and fatal disorder, is a rare cause of pulmonary hypertension. We report the occurrence of PVOD in a female patient with Hasimoto's thyroiditis. This report emphasises that PVOD can co-exist with Hashimoto's thyroiditis and a high index of clinical suspicion is required to confirm the diagnosis of PVOD. PMID- 16255402 TI - Late onset hypoventilation syndrome: is there a spectrum of idiopathic hypoventilation syndromes? AB - We report a case of late onset central hypoventilation syndrome (LO-CHS) with hypothalamic dysfunction (HD) and ganglioneuroma presenting at the age of ten years. LOCHS-HD is now a well-established syndrome; the key is to investigate each child's history and presentation to expeditiously offer the most accurate diagnosis and optimal management. PMID- 16255403 TI - Immune reconstitution syndrome following initiation of antiretroviral therapy in a patient with HIV infection and multidrug-resistant tuberculosis. AB - Paradoxical exacerbation of the signs and symptoms of tuberculosis may occur not only after antituberculosis therapy, but also soon after the initiation of a potent combination of antiretroviral drugs in human immunodeficiency virus (HIV) serpositive patients with tuberculosis. We report a case of immune reconstitution syndrome in response to antiretroviral therapy in a HIV-positive patient on antituberculosis therapy for multidrug-resistant tuberculosis. PMID- 16255404 TI - Mycoplasma pneumonia associated with rhabdomyolysis and the Guillain-Barre syndrome. AB - A 25-year-old housewife who presented with Mycoplasma pneumonia who developed acute respiratory distress syndrome (ARDS) and required assisted ventilation. During her hospital stay, she developed acute renal failure because of rhabdomyolysis and was put on haemodialysis. She also had difficulty in weaning from ventilator because of acute motor-sensory axonal neuropathy (AMSAN) variant of the Guillain-Barre syndrome. The patient was treated with antibiotics and corticosteroids. The patient recovered from both the complications gradually. PMID- 16255405 TI - Guidelines for management of asthma at primary and secondary levels of health care in India (2005). PMID- 16255408 TI - The nursing shortage. PMID- 16255409 TI - While Bundaberg waits. . . PMID- 16255410 TI - Market forces in the private sector or "Where should I work?". PMID- 16255411 TI - Re-birthing: part 2. PMID- 16255412 TI - Keeping it straight: the importance of keeping records. PMID- 16255413 TI - Scientific. Risk of intraoral cancer associated with tobacco and alcohol--a case control study. AB - OBJECTIVES: The study examined the role of tobacco, alcohol consumption and their synergistic effect on the aetiology of intraoral cancer. DESIGN: A hospital based, analytical case-control study. SUBJECTS: Information was collected from 67 intraoral cancer patients attending a cancer unit (cases) and 67 controls attending other clinics in the same hospital. Cases and controls were individually matched for age, gender and ethnicity. METHODS: A structured interview was used to collect the data on items related to smoking status and alcohol consumption. RESULTS: The odds of developing intraoral cancer were 41 times (OR = 4.63; 95% CI: 1.74-12.30) higher for current smokers compared to non smokers. The odds of developing intraoral cancer were eight times higher for past drinkers (OR = 8.59; 95% CI: 2.96-24.92) and current drinkers (OR = 8.54; 95% CI: 3.55-20.50) compared to non-drinkers. Consumers of both tobacco and alcohol were ten times more likely to develop intraoral cancer compared to those who do neither. CONCLUSIONS: The risk of intraoral cancer arising from use of alcohol and tobacco, icant. The findings of the study strongly endorse the rationale behind efforts by the South African government to implement strict tobacco legislation and campaign for responsible drinking. PMID- 16255414 TI - HIV/TB co-infection: literature review and report of multiple tuberculosis oral ulcers. AB - Human immunodeficiency virus/tuberculosis (HIV/TB) co-infected subjects demonstrate enhanced HIV replication and plasma viremia; CD4+ T-cell depletion; morbidity and mortality; and susceptibility to secondary bacterial and fungal infections compared to subjects solely infected with HIV. As the incidence of HIV/TB infection has been increasing, one would have expected to encounter oral lesions of tuberculosis more frequently. However, such oral lesions are uncommon. The lesions usually occur as ulcerations of the tongue. We report an additional case in an HIV/TB co-infected 39 year-old black male, who presented with chronic, painless, multiple oral ulcers, occurring simultaneously on the tongue, bilaterally on the palate and mucosa of the alveolar ridge. Microscopic examination confirmed the presence of chronic necrotizing granulomatous inflammation, with the identification of acid fast bacilli in the affected oral mucosal tissue. Anti-retroviral and anti-tuberculous treatment resulted in the resolution of the oral lesions. Confirmatory histopathological diagnosis following a biopsy is essential to determine the exact nature of chronic oral ulceration in an HIV individual and especially to distinguish between oral squamous cell carcimoma, lymphoma, infection (bacterial or fungal) and non specific or aphthous type ulceration. PMID- 16255415 TI - Orthodontic status and treatment need of 12-year-old children in South Africa using the Dental Aesthetic Index. AB - OBJECTIVE: The aim of this study was to determine the prevalence and severity of malocclusion and orthodontic treatment needs in a sample of 12-year-old South African school children using the Dental Aesthetic Index (DAI), and to assess the relationship between malocclusion and certain socio-demographic variables. METHODOLOGY: The sample comprised 6142, 12-year-old children attending school in seven of the nine provinces of South Africa. For each subject the standard demographic information such as gender, population group, location type and employment status of the parents was collected, after which an intra-oral examination for occlusal status using the DAI was performed. Before the survey, the examiners were calibrated and trained and only examiners with an agreement score greater or equal to 80 per cent were included in the final study. RESULTS: The results showed that 47.7 per cent of the children in the sample presented with good occlusion or minor malocclusion, just over 52.3 per cent presented with identifiable malocclusion, a DAI score larger than 26. Of these, 21.2 per cent had definite malocclusion, 14.1 per cent had severe malocclusion and 16.9 per cent had very severe or handicapping malocclusion. Malocclusion as defined in this study was found to be significantly associated with the different population groups in South Africa, with gender and with dentition stage, but not with the location type or the employment status of parents. CONCLUSIONS: The results of this study show a high prevalence of malocclusion in 12-year-old South African children. The findings provide reliable base-line data regarding the prevalence, distribution and severity of malocclusion as well as useful epidemiological data on the orthodontic treatment needs of 12-year-old children in South Africa. PMID- 16255416 TI - What the general dental practitioner should know about implants (Part 2- Radiology). PMID- 16255417 TI - Sedation corner. Use of herbal drugs. PMID- 16255418 TI - Safety of using nitrous oxide for conscious sedation in asthmatic patients. PMID- 16255420 TI - General practitioner's radiology. Case 35. Primary carcinoma of the floor of the mouth. PMID- 16255421 TI - Position-sensing technologies for movement analysis in stroke rehabilitation. AB - Research has focused on improvement of the quality of life of stroke patients. Gait detection, kinematics and kinetics analysis, home-based rehabilitation and telerehabilitation are the areas where there has been increasing research interest. The paper reviews position-sensing technologies and their application for human movement tracking and stroke rehabilitation. The review suggests that it is feasible to build a home-based telerehabilitation system for sensing and tracking the motion of stroke patients. PMID- 16255422 TI - Evaluation of achievable registration accuracy of the femur during minimally invasive total hip replacement. AB - The aim of the paper was to investigate whether accurate, point-based registration of the intra-operative femur will be achieved within the context of minimally invasive surgery for total hip replacement. Computer tomography images, collected for pre-operative planning purposes, were used to simulate the intra operative registration procedure using algorithms for various levels of measurement noise, different small areas of the femur available to the surgeon, and a limited number of collected data points (20-60). This helped with the choice of design variables to perform in vitro registration on a plastic bone model to validate the procedure, which included a multistart algorithm developed for intra-operative registration. The algorithm minimised the distance between the measured and image-derived surfaces and was able to cope with the presence of multiple local minima given sufficient computational effort, even with realistically large measurement noise. It was found that, if a small patch of the femur was used, accessible by a needle that could at times penetrate thin layers of soft tissue, errors in the order of 1.0 mm in translation and 0.5 degrees in rotation were achievable. PMID- 16255423 TI - Visualisation of intramural coronary vasculature by an imaging cryomicrotome suggests compartmentalisation of myocardial perfusion areas. AB - A technique is presented for the 3D visualisation of the coronary arterial tree using an imaging cryomicrotome. After the coronary circulation of the excised heart was filled with a fluorescent plastic, the heart was frozen and mounted in the cryomicrotome. The heart was then sliced serially, with a slice thickness of 40 microm, and digital images were taken from each cutting plane of the remaining bulk material using appropriate excitation and emission filters. Using maximum intensity projections over a series of images in the cutting plane and perpendicular plane, the structural organisation of intramural vessels was visualised in the present study. The branching end in the smallest visible vessels, which define tissue areas that are well delineated from each other by 1 2 mm wide bands populated only by vessels less than 40 microm in diameter. The technique presented here allows further quantification in the future of the 3D structure of the coronary arterial tree by image analysis techniques. PMID- 16255424 TI - Fractal characterisation of boundary irregularity in skin pigmented lesions. AB - A growing literature shows researcher's interest in fractal analysis, arising from its ability to describe and characterise quantitatively the complexity of several tumour profiles. The aim of the work was to investigate the fractal properties of skin pigmented lesion boundaries. Although melanoma is one of the most aggressive tumours, early detection and a high rate of diagnostic accuracy, followed by timely excision, can allow complete recovery in melanoma patients. A modified approach to fractal dimension estimation was performed that was able to consider, in a data fit procedure, the range in which lesions show fractal properties. Identification of this zone is the most important step towards a correct fractal analysis procedure. The method was checked against a known fractal dimension object (Koch's curve) with an error of 0.007. The fractal dimension was estimated in 110 skin pigmented lesions and showed a significantly increasing linear regression (p < 0.05), from common naevi to naevi with dysplasia to melanomas. This result is important for screening, as it can inform the decision to excise precociously malignant lesions or to avoid unnecessary removal of benign ones. The limitations of the method are discussed. PMID- 16255425 TI - Stroke volume equation for impedance cardiography. AB - The study's goal was to determine if cardiac output (CO), obtained by impedance cardiography (ICG), would be improved by a new equation N, implementing a square root transformation for dZ/dtmax/Z0, and a variable magnitude, mass-based volume conductor Vc. Pulmonary artery catheterisation was performed on 106 cardiac surgery patients pre-operatively. Post-operatively, thermodilution cardiac output (TDCO) was simultaneously compared with ICG CO. dZ/dtmax/Z0 and Z0 were obtained from a proprietary bioimpedance device. The impedance variables, in addition to left ventricular ejection time TLVE and patient height and weight, were input using four stroke volume (SV) equations: Kubicek (K), Sramek (S), Sramek Bernstein (SB), and a new equation N. CO was calculated as SV x heart rate. Data are presented as mean +/- SD. One way repeated measures of ANOVA followed by the Tukey test were used for inter-group comparisons. Bland-Altman methods were used to assess bias, precision and limits of agreement. P< 0.05 was considered statistically significant. CO implementing N (6.06 +/- 1.48 l min(-1)) was not different from TDCO (5.97 +/- 1.41 l min(-1)). By contrast, CO calculated using K (3.70 +/- 1.53 l min(-1)), S (4.16 +/- 1.83 l min(-1)) and SB (4.37 +/- 1.82 l min(-1)) was significantly less than TDCO. Bland-Altman analysis showed poor agreement between TDCO and K, S and SB, but not between TDCO and N. Compared with TDCO, equation N, using a square-root transformation for dZ/dtmax/Z0, and a mass based Vc, was superior to existing transthoracic impedance techniques for SV and CO determination. PMID- 16255426 TI - Diagnosing aortic valve stenosis by correlation analysis of wavelet filtered heart sounds. AB - Traditional auscultation performed by the general practitioner remains problematic and often gives significant results only in a late stage of heart valve disease. Valve stenoses and insufficiencies are nowadays diagnosed with accurate but expensive ultrasonic devices. This study aimed to develop a new heart sound analysis method for diagnosing aortic valve stenoses (AVS) based on a wavelet and correlation technique approach. Heart sounds recorded from 373 patients (107 AVS patients, 61 healthy controls (REF) and 205 patients with other valve diseases (OVD)) with an electronic stethoscope were wavelet filtered, and envelopes were calculated. Three correlations on the basis of these envelopes were performed: within the AVS group, between the groups AVS and REF and between the groups AVS and OVD, resulting in the mean correlation coefficients rAVS, rAVSv.REF and rAVSv.OVD. These results showed that rAVS (0.783 +/- 0.097) is significantly higher (p < 0.0001) than rAVSv.REF (0.590 +/- 0.056) and rAVSv.OVD (0.516 +/- 0.056), leading to a highly significant discrimination between the groups. The wavelet and correlation-based heart sound analysis system should be useful to general practitioners for low-cost, easy-to-use automatic diagnosis of aortic valve stenoses. PMID- 16255427 TI - Feature analysis of pathological speech signals using local discriminant bases technique. AB - Speech is an integral part of the human communication system. Various pathological conditions affect the vocal functions, inducing speech disorders. Acoustic parameters of speech are commonly used for the assessment of speech disorders and for monitoring the progress of the patient over the course of therapy. In the last two decades, signal-processing techniques have been successfully applied in screening speech disorders. In the paper, a novel approach is proposed to classify pathological speech signals using a local discriminant bases (LDB) algorithm and wavelet packet decompositions. The focus of the paper was to demonstrate the significance of identifying the signal subspaces that contribute to the discriminatory characteristics of normal and pathological speech signals in a computationally efficient way. Features were extracted from target subspaces for classification, and time-frequency decomposition was used to eliminate the need for segmentation of the speech signals. The technique was tested with a database of 212 speech signals (51 normal and 161 pathological) using the Daubechies wavelet (db4). Classification accuracies up to 96% were achieved for a two-group classification as normal and pathological speech signals, and 74% was achieved for a four-group classification as male normal, female normal, male pathological and female pathological signals. PMID- 16255428 TI - How to select the elastic modulus for cancellous bone in patient-specific continuum models of the spine. AB - Patient-specific finite element (FE) modelling is a promising technology that is expected to support clinical assessment of the spine in the near future. To allow rapid, robust and economic patient-specific modelling of the whole spine or of large spine segments, it is practicable to consider vertebral cancellous bone in the spine as a continuum material, but the elastic modulus of that continuum material must reflect the quality of the individual vertebral bone. A numerical parametric model of lattice trabecular architecture has been developed for determining the apparent elastic modulus of cancellous bone Ecb in vertebrae. The model inputs were apparent morphological parameters (trabecular thickness TbTh and trabecular separation TbSp) and the bone mineral density (BMD), which can all be measured in vivo, using the spatial resolution of current clinical quantitative computed tomography (QCT) commercial whole-body scanners. The model predicted that Ecb values between 30 and 110 MPa represent normal morphology and BMD of human spinal cancellous bone. The present Ecb to TbTh, TbSp and BMD relationships pave the way for automatic generation of patient-specific continuum FE spine models that consider the individual's osteoporotic or other degenerative condition of cancellous bone. PMID- 16255429 TI - Using recurrent artificial neural network model to estimate voluntary elbow torque in dynamic situations. AB - Muscle modelling is an important component of body segmental motion analysis. Although many studies had focused on static conditions, the relationship between electromyographic (EMG) signals and joint torque under voluntary dynamic situations has not been well investigated. The aim of this study was to investigate the performance of a recurrent artificial neural network (RANN) under voluntary dynamic situations for torque estimation of the elbow complex. EMG signals together with kinematic data, which included angle and angular velocity, were used as the inputs to estimate the expected torque during movement. Moreover, the roles of angle and angular velocity in the accuracy of prediction were investigated, and two models were compared. One model used EMG and joint kinematic inputs and the other model used only EMG inputs without kinematic data. Six healthy subjects were recruited, and two average angular velocities (60 degrees s(-1) and 90 degrees s(-1)) with three different loads (0 kg, 1 kg, 2 kg) in the hand position were selected to train and test the RANN between 90 degrees elbow flexion and full elbow extension (0 degrees). After training, the root mean squared error (RMSE) between expected torque and predicted torque of the model, with EMG and joint kinematic inputs in the training data set and the test data set, were 0.17 +/- 0.03 Nm and 0.35 +/- 0.06 Nm, respectively. The RMSE values between expected torque and predicted torque of the model, with only EMG inputs in the training data set and the test set, were 0.57 +/- 0.07 Nm and 0.73 +/- 0.11 Nm, respectively. The results showed that EMG signals together with kinematic data gave significantly better performance in the joint torque prediction; joint angle and angular velocity provided important information in the estimation of joint torque in voluntary dynamic movement. PMID- 16255430 TI - Influence of gestational age, heart rate, gender and time of day on fetal heart rate variability. AB - From adult data, it is known that numerous factors, such as age, state of the autonomic nervous system, diurnal rhythms or mean R-R interval mRR, influence heart rate variability (HRV). The aim of this study was the examination of the influence of gestational age, mRR, gender and time of day on fetal HRV. The analysis was based on 66 fetal magnetocardiograms (FMCGs) of 22 healthy fetuses between the 16th and 42nd week. FMCGs were recorded for 5 min using a multichannel biomagnetometer. On the basis of the time series of fetal R-R intervals, mRR as well as the standard deviation sdRR, root mean square of successive differences rmssdRR and approximate entropy ApEn were calculated. The influence of gestational age, mRR and gender on sdRR, rmssdRR and ApEn was determined by regression analysis. The relationship between time of day and HRV was evaluated by visual inspection of scatterplots. The logarithmised HRV measures increased significantly with the logarithm of gestational age (regression coefficients: sdRR = 1.28, rmssdRR = 1.12, ApEn = 1.30) and mRR (regression coefficients: sdRR = 0.008, rmssdRR = 0.011, ApEn = 0.012) There was no significant influence of gender. With respect to time of day (between 0800 h and 1800 h), no dependency of the HRV measures was apparent. In summary, when fetal HRV is assessed, it is essential to take gestational age and mRR into account. In contrast, time of day, with respect to daytime, and gender need not be considered. In future studies, the influence of fetal activity state on HRV should be examined. PMID- 16255431 TI - Signal-dependent wavelets for electromyogram classification. AB - In the study, an efficient method to perform supervised classification of surface electromyogram (EMG) signals is proposed. The method is based on the choice of a relevant representation space and its optimisation with respect to a training set. As EMG signals are the summation of compact-support waveforms (the motor unit action potentials), a natural tool for their representation is the discrete dyadic wavelet transform. The feature space was thus built from the marginals of a discrete wavelet decomposition. The mother wavelet was designed to minimise the probability of classification error estimated on the learning set (supervised classification). As a representative example, the method was applied to simulate surface EMG signals generated by motor units with different degrees of short-term synchronisation. The proposed approach was able to distinguish surface EMG signals with degrees of synchronisation that differed by 10%, with a misclassification rate of 8%. The performance of a spectral-based classification (error rate approximately 33%) and of the classification with Daubechies wavelet (21%) was significantly poorer than with the proposed wavelet optimisation. The method can be used for a number of different application fields of surface EMG classification, as the feature space is adapted to the characteristics of the signal that discriminate between classes. PMID- 16255432 TI - Selective activation of small-diameter motor fibres using exponentially rising waveforms: a theoretical study. AB - The present study investigated the possibility of using exponentially rising waveforms for selectively activating small motor fibres in a nerve bundle enclosed by a cuff electrode. Exponentially rising waveforms were studied using models of motor fibres and a volume conductor model. With an exponentially rising waveform (duration: 2 ms, time constant: 1 ms) large (15.5 microm) and small (8 microm) nerve fibres located at the edge of the nerve bundle had a current threshold of 125 microA and 53 microA, respectively. These reversals in the recruitment order of large and small nerve fibres located at the edge of the nerve bundle were observed for exponentially rising waveforms of 2, 4, and 6 ms in duration with time constants of 0.9, 0.6 and 0.6 ms, respectively. Reversals of the same nerve fibres located at the centre of the nerve bundle were observed for exponentially rising waveforms of 4 and 6 ms in duration, with a time constant of 0.6 ms for both waveforms. The underlying mechanism for selective activation of small nerve fibres with exponentially rising waveforms was found to be a combination of a decrease in the size of the local excitations in the centre node due to sodium channel inactivation and blocking of action potentials in large nerve fibres due to their larger difference in the membrane potential of adjacent nodes. The exponentially rising waveforms were compared with both rectangular prepulses and ramp prepulses. The rectangular prepulses were found to be unable selectively to activate small nerve fibres with the volume conductor model and criteria used in the present study, whereas the ramp prepulses performed as well as the exponentially rising waveforms. In conclusion, a novel stimulation paradigm has been proposed that may provide smooth, gradual control of muscle force with minimum fatigue. PMID- 16255433 TI - Methodology for hypoglycaemia detection based on the processing, analysis and classification of the electroencephalogram. AB - Hypoglycaemia (blood glucose level below 3.8 mmol l(-1)) is the most common complication in the treatment of diabetes with insulin and can cause a number of problems. Previous works have shown that hypoglycaemia causes changes in the electroencephalogram (EEG) signal. In this investigation, portable apparatus was developed to record the EEG, and a methodology was implemented, using digital signal processing and artificial neural networks (ANNs), to detect hypoglycaemia. Sixteen EEG recordings were made on eight subjects with diabetes (five male, three female), aged 35 +/- 13.5 years (mean +/- SD), during the day, over periods of 5.7 +/- 2 min. Ten of these recordings (in seven subjects) included periods of normoglycaemia and spontaneous hypoglycaemia. The result of the off-line ANN classification for each of these ten recordings was an overall accuracy rate of 71.3%, sensitivity of 71.1% and specificity of 71.5%. In the classification using four recordings from a single subject, the accuracy was 80.6%, with a sensitivity of 77.8% and a specificity of 83.9%. In the classification using recordings from five different subjects to train the ANN, the obtained accuracy rate was 49.2%, with a sensitivity of 76% and a specificity of 32.5%. The result of the classification in real time, for one subject, was an accuracy rate of 85.2%, with a sensitivity of 60% and a specificity of 100%. In conclusion, the methodology proposed and implemented justifies further studies with the objective of constructing a hypoglycaemia detector system based on the processing and classification of the EEG. PMID- 16255434 TI - Algorithm for ventricular capture verification based on the mechanical evoked response. AB - Automatic pacemaker capture verification is important for maintaining safety and low energy consumption in pacemaker patients. A new algorithm was developed, based on impedance measurement between pacing electrode poles, which reflects the distribution of the conducting medium between the poles and changes with effective contraction. Data acquired during pacemaker implant in 17 subjects were analysed, with intracardiac impedance recorded while pacing was performed in the ventricle at varying energies, resulting in multiple-captured and non-captured beats. The impedance signals of all captured/non-captured beats were analysed using three different algorithms, based on the morphology of the impedance signal. The algorithm decision for each beat was compared with an actual capture or non-capture, as determined from the simultaneous recording of surface ECG. Two of the three algorithms (Z1 and Zn) were based on impedance values, and one (Z'n) was based on the first derivative of the impedance. Z1 was based on a single sample, whereas Z'n and Z'n were based on several samples in each beat. The total accuracy for each was Z1: 43%, Zn: 87%, Z'n: 92%. It was concluded that impedance based capture verification is feasible, that a multiple rather than single sample approach for signal classification is both feasible and superior, and that first derivative analysis with multiple samples (Z'n) provides the best results. PMID- 16255435 TI - Design and evaluation of a handheld impedance plethysmograph for measuring heart rate variability. AB - Heart rate variability (HRV) analysis from 10s ECGs has been shown to be reliable. However, the short examination time warrants a user-friendly system that can be used for ad-hoc examinations without normal preparation, unlike ECG. A handheld device has been developed that can measure ultra-short HRV from impedance plethysmographic recordings of the pulse wave in distal superficial arteries. The prototype device was made user-friendly through a compact, pen-like design and the use of integrated metal electrodes that were especially designed for dry operation. The main signal processing was performed by a digital signal processor, where the discrete heart beats were detected using a correlation algorithm that could adapt to individual pulse wave shapes to account for biological variation. The novel device was evaluated in 20 mainly young volunteers, using 10 s time-correlated ECG recordings as the reference method. Agreement between the two methods in measuring heart rate and root mean square of successive differences in the heart beat interval (RMSSD) was analysed using correlation coefficients (Pearson's R2), mean differences with 95% confidence intervals and 95% limits of agreement, and Bland-Altman plots. The correlation between the two methods was R2 = 1.00 and R2 = 0.99 when heart rate and RMSSD were measured, respectively. The Bland-Altman plots showed suitable agreement between the novel device and standard 10 s ECGs, which was substantiated by 95% limits of agreement of the difference of +/- 0.1 beats min(-1) and approximately +/- 10 ms for heart rate and RMSSD, respectively. Therefore the evaluation showed no significant systematic error of the novel device compared with ECG. PMID- 16255436 TI - Instrument to measure the heat convection coefficient on the endothelial surface of arteries and veins. AB - The primary objective of the paper was to present the design and analysis of an instrument to measure the heat convection coefficient h on the endothelial surfaces of arteries and veins. An invasive thermistor probe was designed to be inserted through the vessel wall and positioned on the endothelial surface. Electrical power was supplied to the thermistor by a constant temperature anemometry circuit. Empirical calibrations were used to relate electrical measurements in the thermistor to the h at the endothelial surface. As the thermal processes are strongly dependent on baseline blood temperature, the instrument was calibrated at multiple temperatures to minimise this potentially significant source of error. Three different sizes of thermistor were evaluated to optimise accuracy and invasiveness, and the smallest thermistors provided the best results. The sensitivity to thermistor position was evaluated by testing the device at multiple locations, varying both depth of thermistor penetration and position along the vessel. Finally, the measurement accuracy of the instrument was determined for the range of h from 430 to 4200 W m(-2)K, and the average error of the reading was 4.9% for the smallest thermistor. Although the instrument was designed specifically for measurements in the portal vein to obtain useful data for current numerical modelling, the device can be used in any large vessel. PMID- 16255437 TI - Two-way communication for programming and measurement in a miniature implantable stimulator. AB - Implantable stimulators are needed for chronic electrical stimulation of nerves and muscles in experimental studies. The device described exploits the versatility of current microcontrollers for stimulation and communication in a miniature implant. Their standard outputs can provide the required selectable constant-current sources. In this device, pre-programmed stimulation paradigms were selected by transcutaneous light pulses. The potential of a programmable integrated circuit (PIC) was thus exploited. Implantable devices must be biocompatible. A novel encapsulation method that require no specialised equipment and that used two classical encapsulants, silicone and Teflon was developed. It was tested for implantation periods of up to four weeks. A novel way to estimate electrode impedance in awake animals is also presented. It was thus possible to follow the evolution of the nerve-electrode interface and, if necessary, to adjust the stimulation parameters. In practice, the electrode voltage at the end of a known constant-current pulse was measured by the PIC. The binary coded value was then indicated to the user as a series of muscle twitches that represented the binary value of the impedance measurement. This neurostimulator has been successfully tested in vitro and in vivo. Thresholds and impedance values were chronically monitored following implantation of a self-sizing spiral cuff electrode. Impedance variations in the first weeks could reflect morphological changes usually observed after the implantation of such electrodes. PMID- 16255439 TI - The public health implications of natural and man-made disasters. PMID- 16255438 TI - Implantable device for long-term electrical stimulation of denervated muscles in rabbits. AB - Although denervating injuries produce severe atrophic changes in mammalian skeletal muscle, a degree of functional restoration can be achieved through an intensive regime of electrical stimulation. An implantable stimulator was developed so that the long-term effects of different stimulation protocols could be compared in rabbits. The device, which is powered by two lithium thionyl chloride batteries, is small enough to be implanted in the peritoneal cavity. All stimulation parameters can be specified over a wide range, with a high degree of resolution; in addition, up to 16 periods of training (10-180 min) and rest (1-42 h) can be set in advance. The microcontroller-based device is programmed through a bidirectional radiofrequency link. Settings are entered via a user-friendly computer interface and annotated to create an individual study protocol for each animal. The stimulator has been reliable and stable in use. Proven technology and rigorous quality control has enabled 55 units to be implanted to date, for periods of up to 36 weeks, with only two device failures (at 15 and 29 weeks). Changes in the excitability of denervated skeletal muscles could be followed within individual animals. Chronaxie increased from 3.24 +/- 0.54 ms to 15.57 +/- 0.85 ms (n = 55, p < 0.0001) per phase in the 2 weeks following denervation. PMID- 16255440 TI - Debunking the 'only 50%' myth: prevalence of established risk factors in New Zealanders with self-reported ischaemic heart disease. AB - OBJECTIVE: To estimate the prevalence of established risk factors for ischaemic heart disease (IHD) in New Zealand adults and compare the prevalence in adults with and without this disease. DESIGN: Data were obtained from the 2002/03 New Zealand Health Survey. Risk factor prevalence was determined by: self-reported doctor diagnosis of high blood pressure, high cholesterol and diabetes; self report of smoking and physical inactivity; and measurement of obesity. Presence of IHD was based on self-report of heart disease (doctor diagnosed at age 25 years or over) together with current medical or past surgical treatment for this disease. Multiple logistic regression was used to determine prevalence rate ratios (PRRs) for males and females separately, adjusting for age, ethnicity and deprivation. RESULTS: The overall prevalence of IHD was 8%. Overall risk factor prevalences were in the range of 20-25% for each of high blood pressure, high cholesterol, smoking, obesity and physical inactivity, and approximately 5% for diabetes. Overall, 94-97% of adults with IHD had at least one risk factor (depending on how smoking was defined). The PRRs of IHD were highest for cholesterol (about 4.5), followed by blood pressure (about 2.3), with all other risk factors around 1.5. PAF estimates indicate that 80-85% of IHD was attributable to the presence of at least one risk factor for all age, gender and ethnic groups. CONCLUSIONS: Established risk factors account for 80-85% of the non-fatal burden of IHD in New Zealand. Limited research resources would be better used to evaluate which interventions are effective and efficient at reducing exposure of all population groups to known risk factors, rather than on identification of additional risk factors. PMID- 16255441 TI - Correlates of habitual snoring and witnessed apnoeas in Busselton, Western Australia. AB - OBJECTIVE: The aim of this study was to identify potential body size, behavioural and respiratory risk factors for habitual snoring and witnessed apnoeas in a general population. METHODS: Correlates of these conditions were studied in a sample of 3,577 adults aged 25-74 years who participated in a comprehensive health survey in Busselton, Western Australia, during 1994/95. Logistic regression was used to assess associations after age and gender adjustment and also in multivariate models. RESULTS: The prevalence of both conditions was higher in men and rose with age. After controlling for age, gender and body mass index no additional body size variable remained significantly associated with witnessed apnoeas, whereas both waist-hip ratio and neck-height ratio remained significantly associated with habitual snoring. Among behavioural variables, smoking showed the strongest association, and among respiratory symptoms, asthma for habitual snoring and bronchitis for witnessed apnoeas had significant independent associations. CONCLUSIONS AND IMPLICATIONS: This study has confirmed obesity and smoking as key determinants of habitual snoring and witnessed apnoeas. It has also shown that a number of measures of obesity are independently related to habitual snoring and that asthma and bronchitis may also play a role, independently of obesity. PMID- 16255442 TI - The costs and benefits of physiotherapy as first-line treatment for female stress urinary incontinence. AB - OBJECTIVE: To evaluate the costs and benefits of physiotherapy for stress urinary incontinence (SUI) in Australia. METHODS: We evaluated the costs and benefits of physiotherapy for the treatment of SUI using outcome data from a prospective multicentre observational study conducted in 1999/2000. Women presenting with SUI to physiotherapists trained in continence management in 35 centres across Australia were recruited into the study. The outcomes of treatment were assessed using subjective, objective and quality-of-life measures at the conclusion of treatment and with 12-month follow-up. The number of treatments in an average episode of care was calculated and adverse events were recorded. RESULTS: Of the 274 consenting subjects, 208 completed an episode of physiotherapy care consisting of a median (IQ range) of five (4-6) visits. The estimated average costs for an episode of ambulatory physiotherapy treatment were dollar 302.40. Based on 'intention to treat' principles, 64% of women were objectively cured. There was a clinically and statistically significant improvement (p < 0.05) in all outcomes after treatment and these were maintained at one-year follow-up. No adverse events were reported. CONCLUSIONS AND IMPLICATIONS: Specialised ambulatory physiotherapy for SUI in Australia is a low-cost, low-risk and effective treatment. These results provide evidence to support international recommendations that physiotherapy should be routinely implemented as first-line treatment before consideration of surgery. This information has important economic implications for planning future health services. PMID- 16255443 TI - Trends in health service use for women in Australia with diabetes. AB - OBJECTIVE: To describe the health, health service use and use of recommended guidelines for care for women in Australia with diabetes. METHODS: Analysis of survey data 1996-99 from the Australian Longitudinal Study on Women's Health, linked with Medicare data for 1997-2001. Participants were 12,338 mid-age women aged 45-50 years in 1996 (1.9% with diabetes) and 10,421 older women aged 70-75 years at Survey 1 in 1996 (8.1% with diabetes). The outcome measures were number of general practice and specialist visits and use of glycosylated haemoglobin (HbAlc), lipids and microalbuminuria tests. RESULTS: Women with diabetes at Survey 1, and those diagnosed by Survey 2, were more likely to have hypertension, heart disease and eyesight problems, have high rates of polypharmacy (four or more medications: mid age 32%, older 64%) and more consultations with general practitioners and specialists than women without diabetes. During 1997-2001, there was a trend for a greater percentage of women with diabetes to have an HbA1c test at least annually (mid age 44%-52%, older age 46%-58%). Rates of testing microalbuminuria and lipids also increased but were far from conforming to guidelines. Having more frequent consultations with a general practitioner was significantly associated with having all three recommended tests. CONCLUSIONS: There is an increasing use of services by women with diabetes, in part due to an increase in compliance with guidelines for the management of diabetes. IMPLICATIONS: Linked health and administrative data provide a means to monitor health service utilisation, adherence to principles for best practice care and issues of equity in care. PMID- 16255444 TI - The individual and health sector costs of asthma: the first year of a longitudinal study in New South Wales. AB - OBJECTIVE: To identify the resources used and the costs incurred by people with asthma for health care and non-health care products and services to manage asthma. METHODS: A prospective, longitudinal study, using self-reported and administrative data, commenced in 2001 in New South Wales (NSW). Data sources included two six-monthly surveys, hospital admission data from NSW Health, and pharmaceutical benefits and Medicare data. A cohort of 245 people with asthma, aged between 5-75 years, was recruited from the general population and from hospital emergency departments. The study measured the use and cost of health care services and products, including alternative therapies and home modifications. Costs to both the health care system and individuals were measured. RESULTS: General practitioner visits and asthma medications were the items of health care most commonly used. Medications were also the largest component of individual costs for health care in terms of the average payment and the number of people facing an out-of-pocket cost, although home equipment and modifications were the most expensive individual items. The distribution of individual costs was highly skewed, ranging from dollar 0 to dollar 4,882 per person per annum (median dollar 89). Admitted hospital care was the largest component of the cost to the health care system. CONCLUSION: While individual costs were not large for the majority, some people faced substantial costs, with the highest of these being for non-health care items. IMPLICATIONS: Asthma management policies may need to incorporate schemes to alleviate the impact of costs for the minority experiencing significant out-of-pocket expenses. PMID- 16255445 TI - Can we generalise to other young people from studies of sexual risk behaviour among university students? AB - OBJECTIVE: Many studies of sexual behaviour and condom use are based on data collected from university students. The aim of this paper is to determine whether first-year university students and their same-age peers have different patterns of sexual behaviour. METHODS: Computer-assisted telephone interviews were completed by a representative sample of 19,307 Australian men and women aged 16 59 years (response rate 73.1%), 920 of whom were aged 17-19 years. Comparisons were made between reports of sexual risk behaviours from first-year university students and reports of the same behaviours from their same-age peers. RESULTS: For female respondents, there were few differences in the sexual behaviour of first-year university students and their same-aged peers. For male respondents, there were some significant differences in the sexual behaviour of first-year university students and their same-aged peers and also different patterns of correlation between measures of sexual behaviour. Socio-demographic characteristics were related to whether 17-19 year-old respondents were first year university students or engaged in other activities. CONCLUSIONS: The findings of studies of the sexual behaviour of university undergraduates should only be generalised to other groups with caution. The socio-demographic characteristics of the student population of a particular institution must be taken into account before generalisation to the broader population can safely be made from studies of single universities. PMID- 16255446 TI - The index of relative socio-economic disadvantage: general population views on indicators used to determine area-based disadvantage. AB - OBJECTIVE: To ascertain general population perceptions of the importance of indicators comprising the Index of Relative Socio-economic Disadvantage (IRSD). METHODS: Data for this study came from a face-to-face Health Omnibus survey of 3,001 residents in metropolitan and country South Australia, conducted in 2003. RESULTS: Overall, respondents viewed the IRSD indicators as important. Of the 14 indicators, seven were seen as important by more than two-thirds of respondents (ranging from 90% perceiving the number of families with children and a low income important to 68% perceiving the number of one-parent families with dependent children as important). Younger respondents and those of lower educational attainment were more likely to perceive the indicators as unimportant, compared with older people. For example, 14% of people aged 15-24 vs. 5% of people aged 55-64 (p < or = 0.001) viewed the indicator 'number of one parent families and dependent children' as unimportant. CONCLUSIONS: While the general population generally recognises the IRSD indicators as important measures of area-based disadvantage, there were systematic age differences in the degree to which individual indicators were deemed important. There was a general lack of support for several indicators (such as proportion of people separated/divorced, houses with no cars). IMPLICATIONS: This research raises the question of which factors are important in representing area-based disadvantage for young people and equally the use of this index when examining variations in the health of young Australians. PMID- 16255447 TI - Is there a difference in health estimates between people with listed and unlisted telephone numbers? AB - OBJECTIVE: To examine the characteristics of people with a silent telephone by demographic and health-related issues and to assess the potential bias in telephone surveys that use the White Pages as the sampling frame. METHOD: A representative annual population health survey of South Australians aged 15 years and over conducted in 1994, 1998, 1999 and 2002. Self-reported information on telephone connection and listings in the White Pages was provided by participants. Questions were included in the 1998 survey on reasons why telephone number was not listed. RESULTS: The rate of unlisted telephone numbers significantly increased (17.3% to 20.2%) in South Australia between 1994 and 2002. People with an unlisted number are more likely to be living in the metropolitan area, single adult households, current smokers, and in the younger age groups. In the multivariate analyses the only health and risk factor variable that was consistently significant over the years was smoking status. The main reason for having an unlisted number was to avoid market research or telemarketing calls (33%). CONCLUSION: Telephone surveys using White Pages samples are suitable to collect information for prevalence on most commonly collected self-reported health conditions and health-related issues. The exception is smoking status. IMPLICATIONS: The use of Whites Pages telephone samples is a suitable method for obtaining population health information because it is efficient when there is limited time and funds. PMID- 16255448 TI - Improving response rates to primary and supplementary questionnaires by changing response and instruction burden: cluster randomised trial. AB - OBJECTIVE: Supplementary questionnaires provide additional information from subgroups, but may have an attritional effect on response rates. We examined the effects of different instruction methods on response rates to a two-part questionnaire. METHODS: The ACT Kindergarten Health Survey comprises a health questionnaire for all school-entry children. A supplementary questionnaire targets children with respiratory symptoms. We cluster-randomised 109 schools in the ACT (4,494 children) to two instruction groups. Group 1 (instruction burden) had instructions to complete the supplementary questionnaire if certain questions in the primary questionnaire were answered. Group 2 (response burden) had instructions to complete both questionnaires irrespective of answers to the primary questionnaire. RESULTS: Instructing all respondents to complete both questionnaires regardless of eligibility resulted in a statistically significantly lower primary questionnaire response rate (82% vs. 87%), but a statistically significantly higher response rate to the supplementary questionnaire (99% vs. 91%). The net effect was a small overall gain (82% vs. 79%) for the response burden group. CONCLUSION: Increasing the response burden had a minor impact on response rate to the primary questionnaire, but increased the response rate for most items to the supplementary questionnaire. IMPLICATIONS: Large surveys may be broken into primary and supplementary questionnaires, if strategies to maximise response rates are used. Questionnaires may need to be modified to take into account the likely attrition on response to either questionnaire resulting from instruction and response burdens. PMID- 16255449 TI - Intraclass correlation coefficients from three cluster randomised controlled trials in primary and residential health care. AB - OBJECTIVE: This paper provides intraclass correlation coefficients (ICCs) for estimation of sample size inflation required in future cluster randomised trials in primary or residential care. METHODS: Three cluster randomised trials were conducted among middle-aged and older adults in primary care and residential care in Australia and New Zealand between 1995 and 2002. Baseline means or proportions, mean change, and ICCs with their standard errors and 95% confidence intervals are reported for outcome variables used in the three studies. The ICCs were estimated from a one-way random effects model using the analysis of variance method. RESULTS: ICCs for quality of life and psychological variables in the primary care studies were low (below 0.018). ICCs for clinical and physical activity variables ranged from 0 to 0.08. ICCs for health and functional status in residential care for the elderly were high, ranging from 0.025 to 0.514. CONCLUSIONS: The magnitude of the intraclass correlation varies with the venue of the trial, the outcome variables used, and the expected effect of the intervention. However, the intraclass correlations provided will be useful for more appropriate planning of residential and primary care-based trials in the future. PMID- 16255450 TI - Moving beyond good intentions: indigenous participation in aboriginal and Torres Strait islander health research. AB - Indigenous participation in every aspect of health research is increasingly recognised as an important element of any research project that aims to improve Indigenous health. Despite the acceptance of its importance, when the concept of 'Indigenous participation' is mentioned, authors are often imprecise as to the nature and purpose of participation, and its relationship to improved health outcomes. This report attempts to bring some clarity to the variety of meanings we might give Indigenous participation in research. For the purposes of stimulating further debate, we identify four distinct, but overlapping, rationales: pragmatic, moral, interventionist, and epistemological. Each has different implications for how Indigenous participation should be implemented and evaluated. More debate on the meanings and purposes of Indigenous participation will contribute to a refined understanding of its potential benefits to health research. PMID- 16255451 TI - Countermeasures to driver fatigue: a review of public awareness campaigns and legal approaches. AB - OBJECTIVE: Driver fatigue accounts for 10-40% of road crashes and is a critical area for public health. As other major road safety issues are more successfully managed, driver fatigue becomes proportionately more important. Both public awareness and legal developments have been slow to reach the same levels as for other road safety risks. The aim of this article is to review countermeasures for non-commercial drivers that are designed to reduce the likelihood of fatigue related crashes through education and legislation. METHODS: This review outlines information from a wide variety of sources including governments, road safety groups and the scientific literature. Educational and legislative approaches are discussed in terms of both their effectiveness and the associated implications for public health. CONCLUSIONS: Areas for improvement in education include personalising the risk to drivers and developing simple metrics for the self assessment of fatigue. Legal implications should be more clearly defined and specific laws are needed to more effectively prosecute fatigued drivers who cause crashes. Additional research is needed to further investigate the efficacy of available countermeasures. IMPLICATIONS: Increasingly, road traffic injury is being discussed more broadly as a public health issue. However, the specific issue of driver fatigue still receives less attention than other main causes of road crashes, despite making a significant and comparable contribution to crash rates. Countries such as Australia and New Zealand have a responsibility to counter driver fatigue, as well as other causes of road crashes, and to further pursue improvements for the benefit of public health. PMID- 16255452 TI - Public water fluoridation and dental health in New South Wales. AB - OBJECTIVES: To evaluate whether access to fluoridated public water in New South Wales (NSW) is related to both a reduction in caries experience within NSW regions and to better dental health for disadvantaged children. METHODS: Cross sectional population data on children attending the School Dental Service in NSW in 2000 were used to calculate and compare the number of decayed, missing and filled teeth (dmft/ DMFT) across areas of differing availability of fluoridated water within NSW Area Health Service (AHS) regions. Analyses were also undertaken looking at differences in caries between optimally fluoridated and non fluoridated communities across strata of socio-economic disadvantage and by Indigenous status. RESULTS: A total sample of 248,944 children aged 3-15 years was obtained. Caries experience in the deciduous dentition of 5-6 year-olds and the permanent dentition of 11-12 year-olds was significantly lower for children in fluoridated areas than nonfluoridated areas in six of the eight AHSs and six of the 10 AHSs respectively where comparisons could be made. Children living in fluoridated areas had lower caries experience than children living in nonfluoridated areas, regardless of socio-economic disadvantage. Both Indigenous and non-Indigenous children had reduced caries experience in fluoridated compared with non-fluoridated areas. CONCLUSIONS: Water fluoridation was found to be related to significantly reduced caries experience in the majority of AHSs where comparisons could be made, and to benefit all socio-economic strata of the community. IMPLICATIONS: Water fluoridation should be extended to those areas of NSW that are yet to benefit from this successful caries preventive public health initiative. PMID- 16255453 TI - Disease surveillance and response: update on the biological weapons convention. AB - At a meeting in Geneva in December 2004, the member States of the Biological Weapons Convention discussed disease surveillance and response in the context of alleged use of biological weapons, disease outbreaks deemed 'suspicious' and naturally occurring outbreaks. The meeting itself had no mandate to direct States to undertake specific actions regarding these issues. However, the discussions indicated that the problem of biological weapons may be shifting away from the traditional arms control paradigm and towards broader notions of disease-based threats to national and international security. In an era of emerging and re emerging infectious diseases, security analysts are beginning to recognise that strong public health surveillance and response mechanisms are vital defences during disease outbreaks, whether deliberately caused or of natural PMID- 16255454 TI - SF-36: not the tool to monitor the health of subpopulations within the Queensland women's prison system. PMID- 16255455 TI - Tobacco use among public health professionals in Beijing: the relationship between smoking and education level. PMID- 16255456 TI - NSPs and the media: a case study of balanced and accurate reporting? PMID- 16255457 TI - The public health 'Bledisloe Cup': physical activity prevalence differences between New Zealand and Australia. PMID- 16255458 TI - An affirmation of fallibility. PMID- 16255459 TI - Dental digital radiography: more than a fad, less than a revolution. PMID- 16255460 TI - Practice-based research. PMID- 16255461 TI - Justifiable criticism. PMID- 16255462 TI - Filmless imaging: the uses of digital radiography in dental practice. AB - BACKGROUND: As use of digital radiography becomes more common, many dentists are wondering if and how they can replace conventional film-based imaging with a digital system. This article briefly describes the different technologies used for digital radiography in dentistry. The article provides general practitioners with a broad overview of the benefits and limitations of digital radiography to help them understand the role the technology can play in their practices. OVERVIEW: The two technologies now available are solid-state systems and phosphor plate systems. Each has its strong points, and the choice of which to use depends on the type of dental practice. Image processing improves the diagnostic quality of the radiographic information. Advanced image-processing techniques, such as subtraction radiography, are available for specialized clinics. CONCLUSIONS AND CLINICAL IMPLICATIONS: Digital radiography no longer is an experimental modality. It is a reliable and versatile technology that expands the diagnostic and image sharing possibilities of radiography in dentistry. Optimization of brightness and contrast, task-specific image processing and sensor-independent archiving are important advantages that digital radiography has over conventional film-based imaging. PMID- 16255463 TI - Specialty and sex as predictors of depression in dentists. AB - BACKGROUND: Stress, burnout, substance abuse and suicide among dentists have been studied, yet no study in the United States has specifically addressed depression in e dentists. The objective of the authors' study was to determine if sex and dental specialty were correlated with depression in dentists. METHODS: The authors conducted a survey of a sample of dentists chosen randomly from the American Dental Association's mailing list of member dentists. The survey, stratified by sex and specialty, resulted in 560 responses, for a 53 percent response rate. The authors used the Zung Self-Rating Depression Scale to measure depression. The authors examined the respondents' sex, age, number of children, marital status, specialty, practice type, location of practice, years in practice and hours worked per week. RESULTS: The rate of depression in the overall sample was 9 percent. Sex was associated with depression (P < .001), but specialty was not. However, multiple regression analysis found that sex was significantly related to depression in only two specialties: periodontics and pediatric dentistry. Overall, the regression model explained an unimpressive 6 percent of the variance in depression scores. The most important finding of the study was that only 15 percent of depressed dentists were receiving treatment. CONCLUSIONS: The survey results showed that only female pediatric dentists and periodontists were more depressed than their male counterparts. None of the other variables studied contributed significantly to the understanding of depression in dentists. Depressed dentists, like other depressed people, tend not to seek treatment. CLINICAL IMPLICATIONS: Depression and serious depression occur among dentists, and much of it is untreated. Because depression is harmful to dentists and raises quality-of-care issues, they should be educated to help them recognize depression and encouraged to seek treatment. PMID- 16255464 TI - Oral health findings for HIV-infected adult medical patients from the HIV Cost and Services Utilization Study. AB - BACKGROUND: The HIV Cost and Services Utilization Study (HCSUS) was conducted by a consortium of private and government institutions centered at the RAND Corp. to provide national estimates of adult medical patients who are HIV-positive. This article presents descriptive oral health findings from that study. METHODS: The National Opinion Research Center (NORC) conducted four interviews of a nationally representative sample of adults with HIV who made a medical visit for regular care in early 1996. This article uses data from the second interview conducted between December 1996 and July 1997. The authors constructed analytical weights for each respondent so the 2,466 interviewees represented a population of 219,700. RESULTS: Most adult medical patients with HIV rated their oral health as at least "good," but 12 percent (representing a population of 25,300) rated it as "poor." Xerostomia was the most commonly reported symptom (37 percent) to arise in the time since the previous interview. Twenty-nine percent had a dental benefit under Medicaid and 23 percent had private insurance. Eighteen percent had not revealed their HIV status to the dentist they usually saw. CONCLUSIONS: National data on adult medical patients with HIV provide a context for local or convenience sample studies and can help give direction to public health and public policy programs directed to the oral health needs of this population. CLINICAL IMPLICATIONS: The attitudes and beliefs of adult HIV patients should be taken into account in the creation of community health education programs and continuing education for dentists. Medicaid programs should include adult dental benefits. PMID- 16255465 TI - National estimates of out-of-pocket dental costs for HIV-infected users of medical care. AB - BACKGROUND: Out-of-pocket costs for U.S. dental care in 1996 were dollar 157 per person at the poverty level and dollar 229 for people with higher incomes. This article examines out-of-pocket expenditures for dental care in HIV-infected patients who took part in the HIV Cost and Services Utilization Study (HCSUS), conducted by a consortium of private and government institutions centered at the RAND Corp. METHODS: The HCSUS used a probability sampling design. The authors used a weighted sample of 2,466 HCSUS respondents to estimate the national population of HIV-infected users of medical care. The patients were asked to report how much they had spent on their dental care in the preceding 12 months, including payments made by them, their family or their friends--but not by insurance companies--for their dental care. RESULTS: In 1996, 135,000 HIV infected subjects spent dollar 20.5 million on dental care, averaging dollar 152 per user. Whites spent dollar 220, African-Americans dollar 55 and Hispanics dollar 101. People receiving dental care from private dentists spent dollar 232 compared with dollar 7 spent by those who received care in AIDS clinics. CONCLUSIONS: More than one-half of the HIV-infected users of medical care identified private dentists as their source of dental care and spent the vast majority of the dollar 20.5 million. The remaining subjects identified public dental programs as their source of care and had low expenditures. The explanation for these low expenditures is the subsidization of public programs and the likelihood that fewer and less costly services are provided by such programs. PRACTICE IMPLICATIONS: Dentists should be aware of out-of-pocket funds spent by the HIV-infected population in private practices and public programs. Any policy change reducing public funding may result in higher out-of-pocket costs for disadvantaged groups or in increasing disparities in access to dental services. PMID- 16255466 TI - Ensuring the oral health of patients with HIV. PMID- 16255467 TI - Effectiveness of computerized delivery of intrasulcular anesthetic in primary molars. AB - BACKGROUND: Pain measures associated with computerized delivery of intrasulcular anesthestic have not been reported. The authors evaluated a computerized delivery system for intrasulcular (CDS-IS) anesthesia in primary molars. METHODS; The study population consisted of children aged 2 to 13 years who received CDS-IS injections, 159 in mandibular molars and 48 in maxillary molars. Children were treated by one of three modes of behavioral management: behavior modification (BM) only, inhalation of nitrous oxide (N2O) in addition to BM or intrarectal sedation. Variables evaluated included the subjective perception of the child's well-being before and after administration of the anesthetic, the child's pain behavior during anesthetic administration, effectiveness of the anesthetic during dental treatment, incidence of reported postoperative dental pain (PDP) and analgesic use after the CDS-IS injections. RESULTS: The effectiveness of CDS-IS anesthesia in mandibular molars was 97 percent, 92 percent, 63 percent and 71 percent for restorations, preformed stainless steel crowns, extractions and pulpal therapies, respectively (mean effectiveness, 89 percent). The effectiveness of CDS-IS anesthesia in maxillary molars was 96 percent, 50 percent, 92 percent and 78 percent, respectively (mean effectiveness, 90 percent). CDS-IS was less effective in children aged 2 to 4 years who received sedation than it was in older children. The authors found no differences between children's subjective self-reports of well-being before and after anesthetic administration, between the sexes and/or between modes of behavioral management (that is, BM or N2O). Most children exhibited low pain-related behavior during anesthetic administration, with no differences between boys and girls. The overall incidence of PDP was 31.4 percent; 64.9 percent of these patients received pain-relieving medications as a result, with no correlation to age, tooth treated, effectiveness of anesthesia or type of treatment. CONCLUSIONS: CDS IS is effective for anesthetizing primary molars, mainly for amalgam, resin-based composite and stainless steel crown restorations. PMID- 16255469 TI - What has happened to conservative tooth restorations? AB - There is an obvious tendency in dental practice to place more full crowns than may be necessary. This article has discussed some of the reasons for this increased use of full crowns, and for a reduction in the use of full crowns. I have supported the desirability of placing more conservative restorations, and I have discussed the financial and longevity implications offered by more conservative restorative dentistry. PMID- 16255468 TI - Clinical diagnosis of recurrent caries. AB - BACKGROUND: The clinical diagnosis of recurrent caries is the most common reason for replacement of all types of restorations in general dental practice. Marked variations in the diagnosis of the lesions have been reported. The prevention of recurrent lesions by the use of fluoride-releasing restorative materials has not been successful. TYPES OF STUDIES REVIEWED: The author focused on practice-based studies in the literature. These studies are not scientifically rigorous, but they reflect "real-life" dental practice. Few experimental studies on recurrent carious lesions in vivo have been reported, but bacteriological studies indicate that the etiology is similar to that of primary caries. RESULTS: Recurrent carious lesions are most often located on the gingival margins of Class II through V restorations. Recurrent caries is rarely diagnosed on Class I restorations. The diagnosis is difficult, and it is important to differentiate recurrent carious lesions from stained margins on resin-based composite restorations. Over-hangs, even minute in size, are predisposed to plaque accumulation and the development of recurrent caries. The development of recurrent lesions is unrelated to microleakage. CLINICAL IMPLICATIONS: As recurrent carious lesions are localized and limited, alternative treatments to restoration replacement are suggested. Polishing may be sufficient. If not, exploratory preparations into the restorative material adjacent to the localized defect can reveal the extent of the lesion. Such explorations invariably show that the lesion does not progress along the tooth-restoration interface. The defect, therefore, may be repaired in lieu of being completely replaced. Repair and refurbishing of restorations save tooth structure. These simple procedures also increase the life span of the restoration. PMID- 16255470 TI - Clinical implications of cyclo-oxygenase-2 inhibitors for acute dental pain management: benefits and risks. AB - BACKGROUND; Cyclo-oxygenase-2 inhibitors (COX-2i) demonstrate analgesic efficacy for patients who require gastrointestinal safety. The authors discuss the potential benefits and risks of these novel, but expensive, analgesics when used in dentistry. METHODS: The authors conducted a MEDLINE search focused on the subject headings of common analgesic drugs and COX-2i, using peer-reviewed journals limited to the English language. They selected for review 127 articles that met the criteria. They also tried to identify any randomized controlled trials pertinent to dentistry and indicative of evidence-based medicine. RESULTS. When comparing COX isoforms (COX-1 and COX-2), the authors found that overlapping and mutually exclusively properties coexist. COX-2i originally were developed to minimize interference with the gastroprotective properties of the COX-1 isoform, while selectively preventing prostanoid synthesis expressed solely at sites of bodily trauma or other inflammation. COX-2i were found to provide pain relief equal to or slightly exceeding that offered by many mild narcotics. They may avoid some of the serious side effects that can occur with even short-term use of nonselective nonsteroidal anti-inflammatory drugs. CONCLUSIONS: The pharmacodynamics of COX-2i reveal an agent that includes analgesic, anti inflammatory and gastroprotective properties but also allows for an undesirable disruption of the delicate hemodynamic balance. CLINICAL IMPLICATIONS: Symptomatic and asymptomatic gastroparietic patients who do not have severe cardiovascular, cerebral or renal ischemic disease benefit from use of COX-2i. Long-term use of these agents in medically compromised patients may prove disastrous. PMID- 16255471 TI - Continuing dental education requirements for relicensure in the United States. AB - BACKGROUND: Mandatory continuing education (CE) is an almost universal requirement for relicensure of dentists in the United States. In recent years, computer-based methods have become more widely adopted as a vehicle for earning CE credits. To obtain a comprehensive overview of CE requirements in the United States, the authors conducted a survey on how computer-based CE methods are regulated. METHODS: The authors reviewed the dental statutes of 50 states and the District of Columbia regarding the license renewal period, required number of CE hours, limitations on clinical and nonclinical subjects, on-site versus independent-study courses, and other requirements regarding CE. RESULTS: The authors found that 45 states and the District of Columbia mandate CE for relicensure at this time. Most dentists were required to complete approximately 20 hours per year. Ten states specified a minimum number of clinical hours, 17 states limited nonclinical hours, and seven states placed constraints on both clinical and nonclinical CE. Sixty-five percent of states limited the number of CE credits that could be accumulated through independent study. Specific requirements for computer-based methods of earning CE credits were absent in general. CONCLUSIONS: State licensing boards across the United States have implemented comprehensive requirements for CE. Few, if any, provisions addressed computer-based methods of earning CE credits. PRACTICE IMPLICATIONS: Given the increasing adoption of computer-based methods of earning CE credits, state dental boards and accrediting agencies may want to consider steps to improve the quality of computer-based methods of earning CE credits. PMID- 16255472 TI - The dental safety net in Connecticut. AB - BACKGROUND: Many poor, medically disabled and geographically isolated populations have difficulty accessing private-sector dental care and are considered underserved. To address this problem, public- and voluntary-sector organizations have established clinics and provide care to the underserved. Collectively, these clinics are known as "the dental safety net." The authors describe the dental safety net in Connecticut and examine the capacity and efficiency of this system to provide care to the noninstitutionalized underserved population of the state. METHODS: The authors describe Connecticut's dental safety net in terms of dentists, allied health staff members, operatories, patient visits and patients treated per dentist per year. The authors compare the productivity of safety-net dentists with that of private practitioners. They also estimate the capacity of the safety net to treat people enrolled in Medicaid and the State Children's Health Insurance Program. RESULTS: The safety net is made up of dental clinics in community health centers, hospitals, the dental school and public schools. One hundred eleven dentists, 38 hygienists and 95 dental assistants staff the clinics. Safety-net dentists have fewer patient visits and patients than do private practitioners. The Connecticut safety-net system has the capacity to treat about 28.2 percent of publicly insured patients. CONCLUSIONS: The dental safety net is an important community resource, and greater use of allied dental personnel could substantially improve the capacity of the system to care for the poor and other underserved populations. PMID- 16255473 TI - Forming an interdisciplinary team: a key element in practicing with confidence and efficiency. PMID- 16255474 TI - 'Readily achievable': a look at public accommodation requirements under the Americans with Disabilities Act. PMID- 16255475 TI - For the dental patient ... methamphetamine use and oral health. PMID- 16255477 TI - "We want a bursary that will let us live rather than exist'. PMID- 16255478 TI - You'll never have it so good again. PMID- 16255479 TI - Sharp thinking. AB - Nurses are the healthcare professionals most at risk from neeldestick and sharps injuries, according to the latest figures from the RCN's EPINet project. During 2004, 1,990 injuries were reported from the 20 self-selecting participating sites. Nurses sustained 43 per cent of these injuries. Disposable syringes and needles were the most common equipment to cause injury. Although discouraged, 4 per cent of injuries occurred during recapping of needles. Just under a third of injuries were sustained by staff other than the original user of the sharp. PMID- 16255480 TI - Eye on the needle. Interview by Lynne Pearce. AB - Sally O'Toole has been a driving force in the introduction of safer cannulas and reducing the number of sharps injuries in her large NHS trust. Sharps injuries featured among the top three categories of injuries in the trust. Getting key stakeholders on board was crucial to the introduction of new cannulas. Working with the manufacturer meant product costs were not increased. An extensive programme of staff training was undertaken. Changing the way sharps injuries are reported should allow better monitoring. PMID- 16255482 TI - Drive thru spirituality. PMID- 16255481 TI - Hepatitis C nightmare. Interview by Charlotte Alderman. AB - Sustaining a needlestick injury resulted in nurse Dianne Wright contracting hepatitis C infection in 2003. Despite 24 weeks of gruelling treatment, she continued to work. A blood test taken at six weeks after the injury confirmed hepatitis C. Treatment with ribavirin and interferon lasted 24 weeks. Hepatitis C virus was undetectable half way through treatment. Viral clearing was confirmed at the end of treatment and six and 12 months later. PMID- 16255483 TI - Mortal thoughts. PMID- 16255484 TI - Who will protect the innocent from smoke? PMID- 16255485 TI - Making the most of research opportunities for nursing. AB - The Department of Health (2005) consultation document Best Research for Best Health: A New National Health Service Research Strategy offers both opportunities for the future development of nursing research and threats to its future. Lessons from the history of health research reforms in the UK suggest that it will take time for any benefits to become obvious and that only some members of the research community will receive funding. In the past few years the quality, potential leadership and skills base for nursing research have shown unprecedented improvements. Combined with a professional understanding of nurses' careers in both practice and education, the proposed new strategy could work to the advantage of nurses and the patients who rely on their expertise. It is, however, essential that the experience and ambitions of as many nurses as possible now feed into this consultation process. PMID- 16255486 TI - Evaluating a respiratory intermediate care team. AB - This article examines the work of a respiratory intermediate care team. Findings from an audit and a patient satisfaction postal questionnaire are presented to assess the effectiveness of the team's work and evaluate the effect of specialist nursing teams on hospital bed days and patient preference. From the evaluation it is apparent that the respiratory intermediate care team is an effective means of saving hospital bed days through prevention of admission and early discharge in patients with chronic respiratory disease. The results of the patient satisfaction postal questionnaire show that the majority of patients find hospital-at-home style care favourable because they feel well-supported at home to self-manage their condition. This outcome is important because it indicates that specialist nursing teams in primary care are valuable to patients and save the NHS money. PMID- 16255487 TI - Utility and limitations of genetic testing and information. AB - This article introduces some of the issues involved in genetic testing and information, particularly the utility and limitations of such testing. Psychosocial and ethical issues that may arise in this area are also discussed. The aim of this article is to stimulate readers' awareness of and insight into these matters in the hope that practitioners will examine and reflect on the applicability of these to their area of practice. PMID- 16255488 TI - Infection control: the environment and service organisation. AB - Healthcare-associated infection is one of the major challenges to providing high quality health care. Policy makers in the United Kingdom are emphasising initiatives to increase compliance with hand hygiene protocols because most cross infection occurs via hands. The contribution of the environment to risks of infection has received less attention, despite media reports of dirty hospitals. However, the environmental contribution may be greater than previously considered. Changes in the way health care is delivered, especially the movement of patients and staff between different hospitals and wards, may also contribute to the difficulty of controlling infection. PMID- 16255489 TI - Consent. PMID- 16255490 TI - Positive turn. PMID- 16255491 TI - Help yourself. PMID- 16255492 TI - Happy returns. PMID- 16255493 TI - Listening to learners' concerns and complaints. PMID- 16255494 TI - Salvage surgery after radiation failure in squamous cell carcinoma of the larynx. AB - AIM AND BACKGROUND: To investigate patients treated with radiotherapy (RT) for laryngeal squamous cell carcinoma (SCC) in order to estimate the recurrence rate and treatment used as salvage surgery. The survival rate in the group of patients treated with salvage surgery was compared to that of patients who had chemotherapy rather than surgery or who refused any treatment. METHODS: From 1989 to 1999, 185 patients came to our institution for laryngeal SCC. All of them underwent RT as primary treatment. Only patients with a minimum of three years follow-up (n = 143) were included in the study group. RESULTS: The 143 cases included 22 loco-regional recurrences (15.3%) during the minimum three years of follow-up. Recurrence was observed in the larynx in eighteen cases (81.8%), in cervical nodes in one case (4.55%) and in both the larynx and cervical nodes in one case (4.55%). There was peristomal recurrence in two cases (9.1%). Recurrence was observed after an average of 16.3 months. Fourteen patients (63.6%) out of the twenty-two cases of recurrence underwent salvage surgery. Surgery was not performed on the remaining eight patients (36.4%). The global survival rate was 92.3% after three years and 66.6% after five years in the group of patients treated with surgery. The actuarial survival rate was 100% after three years and 83.3% after five years. The global and actuarial survival rate was 20% after three years and 0% after five years in the group of patients who received chemotherapy rather than surgery or who refused any kind of treatment. CONCLUSION: The RT seems to play an important role in the loco-regional control of laryngeal SCC (especially in glottic T1). Salvage surgery for recurrence results in a good survival rate. PMID- 16255495 TI - Skin metastases from a head and neck carcinoma: a prognostic factor? AB - BACKGROUND: Distant metastases in head and neck cancer are often to the lung, and rarely to the skin sites. METHOD: To ascertain that a skin metastasis indicates a poor prognosis, a review was conducted of patients with head and neck cancer. RESULTS: Three out of seven patients had evidence of local recurrence at the time of development of the skin metastases, and three out of seven had evidence of distant metastases. The average survival time was 1 month after the development of skin metastases. CONCLUSION: Skin metastases may represent the first clinical evidence of impending recurrence or distant metastasis. The development of skin metastasis is associated with a poor prognosis. Current treatment options are limited in their efficacity. PMID- 16255496 TI - Gastroesophageal reflux in patients with laryngeal disorders. AB - INTRODUCTION AND AIM: The aim of this study is to use anamnesis and laryngeal examination to investigate the occurrence of the gastroesophageal reflux disease GERD (with symptoms of regurgitation and heartburn, signs of oesophagitis, and pathological pH monitoring) among patients with suspected laryngopharyngeal reflux. MATERIALS AND METHODS: One hundred and sixty-nine patients with laryngopharyngeal reflux indicated by anamnesis and laryngeal examination were evaluated retrospectively with endoscopy and 24-hour double-probe pH monitoring. RESULTS: Combining endoscopy and pH monitoring made possible a diagnosis of GERD in 130 patients (77%). Of these patients, 58% had erosive oesophagitis and 84% had pathological oesophageal acid exposure. Of the patients with suspected laryngopharyngeal reflux and with established GERD, 49% reported gastrointestinal symptoms like heartburn or acid regurgitation. Erosive oesophagitis was confirmed in 45% of patients: grade 1 in 60 (35%), grade 2 in 10 patients (6%), and grades 3 and 4 in 3 patients each (2%). Distal oesophageal pH monitoring was pathological in 65% of patients. Of these, 93% had upright pathological acid exposure. Proximal oesophageal acid exposure was pathological in 39% of patients. CONCLUSIONS: Reflux can be suspected when a patient presents typical laryngeal symptoms and signs. The majority of these patients will not have typical GERD symptoms like heartburn, and endoscopic oesophagitis, mostly grade 1, is present in only a subset of patients with LPR. Oesophageal pH monitoring is the most appropriate test to demonstrate pathological oesophageal acid exposure. PMID- 16255497 TI - Genetic testing for hearing impairment. AB - For some patients, genetic testing can reveal the etiology of their hearing impairment, and can provide evidence for a medical diagnosis. However, a gap between fundamental genetic research on hereditary deafness and clinical otology emerges because of the steadily increasing number of discovered genes for hereditary hearing impairment (HHI) and the comparably low clinical differentiation of the HHIs. In an attempt to keep up with the scientific progress, this article enumerates the indications of genetic testing for HHI from a clinical point of view and describes the most frequently encountered HHIs in Belgium. Domains of recent scientific interest, molecular biological aspects, and some pitfalls with HHIs are highlighted. The overview comprises bilateral congenital hearing loss, late-onset progressive high frequency hearing loss, progressive bilateral cochleo-vestibular deficit, and progressive low frequency hearing loss. Also, several syndromal forms of HHI are summarized, and the availability of genetic tests mentioned. Finally, the requirements for successful linkage analysis, an important genetic research tool for localizing the potential genes of a trait on a chromosome, are briefly described. PMID- 16255498 TI - Regulation of expression of galectin-7 in human nasal polyps by budesonide. AB - BACKGROUND: Nasal polyposis is a model for the study of inflammatory processes. We analyzed the expression of galectin-7, a growth regulator, in surface epithelium, glandular epithelium, and connective tissue in human nasal polyps, and examined the effect of the glucocorticoid budesonide on its expression in human nasal polyps ex vivo. METHODS: Using quantitative, computer-assisted microscopy and immunohistochemistry, we measured galectin-7 expression in nine nasal polyps obtained by surgical resection. Five polyps came from allergic patients and four came from non-allergic patients. RESULTS: Galectin-7 was expressed in all three polyp tissues analyzed. Treatment of polyps from allergic and non-allergic patients with 50 ng/ml budesonide increased the extent of galectin-7 expression in the connective tissue (p = 0.01). Conversely, budesonide at this concentration did not apparently affect galectin-7 expression in glandular epithelium; only a slight decrease in the percentage of the galectin-7 immunopositive cells was observed. In the surface epithelium of nasal polyps from non-allergic patients, the percentage of galectin-7-immunopositive cells was decreased (p = 0.03) by treatment with 250 ng/ml budesonide. In nasal polyps from allergic patients, this percentage was increased by treatment with 50 ng/ml budesonide (p = 0.0001). CONCLUSIONS: These data are consistent with a role for galectin-7 in the regulation of cell growth through a pro-apoptotic effect. Galectin-7 expression coincides with the degree of epithelial stratification, and is subject to upregulation in the connective tissue in response to treatment with 50 ng/ml budesonide. Budesonide modulates galectin-7 expression differently in the surface epithelia of polyps from allergic and non-allergic patients. PMID- 16255499 TI - Laryngeal leiomyosarcoma. AB - Laryngeal leiomyosarcoma is rare. The authors report the case of a 48-year-old patient who presented with progressive hoarseness. This symptomatology was associated with a laryngeal tumour. Immunohistopathological study indicated a leiomyosarcoma. Acute dyspnoea resulted in an urgent tracheostomy, followed by total laryngectomy without neck dissection later. In the light of this observation and a literature review, histopathological characteristics, clinical aspects and therapeutic indications will be discussed for this localisation. PMID- 16255500 TI - Syndrome of inappropriate antidiuretic hormone secretion due to recurrent oral cancer. AB - Hyponatremia is a common metabolic disorder in clinical practice and is associated with significant morbidity and mortality, especially among the elderly. Hyponatremia resulting from the syndrome of inappropriate secretion of antidiuretic hormone (SIADH) has been reported in association with neoplasia (including a few reports in patients with head and neck malignancies) and may represent a paraneoplastic condition. Patients with SIADH present with signs and symptoms that cannot be explained by the primary tumour mass effect or its metastases. We describe a 67-year-old male patient with oral squamous-cell carcinoma of recent recurrence admitted because of symptomatic severe hyponatremia resulting from SIADH and discuss the principles of the diagnostic approach and appropriate management. PMID- 16255501 TI - Endoscopic view and MRI of a Thornwaldt's cyst of the nasopharynx. AB - A Thornwaldt's cyst is an uncommon nasopharyngeal lesion that develops from the remnant of the primitive notochord. A 65-year-old man with a Thornwaldt's cyst is presented in this case report. The patient was diagnosed by rigid nasal endoscopy and magnetic resonance imaging (MRI). These cases are infrequently presented in the English journals. Our study suggested that endoscopic and MRI examinations of the nasopharynx were a simple, rapid, and useful procedure for the diagnosis of the Thornwaldt's cysts. PMID- 16255502 TI - Metastasis of a small cell carcinoma from undetected origin to the pituitary gland--the otolaryngologist's view. AB - Tumour metastasis to the anterior pituitary-hypothalamic area is rare. We present a patient who had severe headache, bitemporal quadrant hemianopsia and an expanding mass in the sella turcica as revealed by MRI. Partial resection via a transsphenoidal approach was performed and postoperative radiation therapy was initiated. Immunohistochemical investigation identified the tumour as a metastatic small cell carcinoma whose primary site remained undetected for more than 12 months despite repeated oncological evaluations. We reviewed the literature on metastatic disease of the pituitary gland. PMID- 16255503 TI - [Nutritional problems in the "silver generation". The hungry elderly...]. PMID- 16255504 TI - [Nutritional references for dementia patients. "Finger food" and fishes in the aquarium]. PMID- 16255505 TI - [Vitamin D, folic acid and cobalamin deficiency. Underestimated risk]. PMID- 16255506 TI - [Patient complains exclusively about aura phenomena. Can there also be migraine without headache? (interview by Christine Vetter)]. PMID- 16255507 TI - ["In near drowning every second counts". No lengthy search for a pulse]. PMID- 16255508 TI - [Not underestimating hypothermia]. PMID- 16255509 TI - [Silent gallbladder calculi. Leave them alone or operate?]. PMID- 16255510 TI - [Women patients with heart manifestations are medically underserved. Stress test women often!]. PMID- 16255511 TI - [External manifestations of internal diseases. Look out for skin involvement]. PMID- 16255512 TI - [Typical skin changes associated with internistic disorders]. PMID- 16255513 TI - [Diabetes mellitus: Cutaneous and mucosal marker lesions]. AB - Diabetes mellitus may be associated with dermatological lesions affecting the skin and mucous membranes. In some cases they may even provide the physician with a first indication that the patient may be suffering from diabetes. Typical examples include necrobiosis lipoidica, granuloma annulare, acanthosis nigricans and vitiligo. PMID- 16255514 TI - [Inflammatory bowel diseases--associated lesions affecting the skin and mucous membranes]. AB - Crohn's disease or ulcerative colitis may be preceded, followed or accompanied by a variety of extra-intestinal diseases affecting the skin and mucous membranes. Amongthe most important of these associated lesions are, for example, erythema nodosum and pyoderma gangrenosum. PMID- 16255515 TI - [Elderly patients behind the wheel]. AB - Elderly drivers exhibit typical deficits when called upon to deal with complicated traffic situations. These are based on psychophysical limitations associated with old age, multimorbidity, and potential undesired drug interactions. Of particular importance in such situations are critical self reflection with regard to perceived deficits, and a qualified evaluation bythe physician of any reduction in competence that is of relevance to road safety. However, many patients ignore illness- or age-related deteriorations in performance. In such cases, neutral and low-level preventive counseling and training options that can be integrated into the medical counseling process may be effective. PMID- 16255516 TI - [Muscular pain in viral infections]. PMID- 16255517 TI - [How many diabetic patients in your practice? Important references for regress defense]. PMID- 16255518 TI - [Regress prevention. Attention, trap in consultation need]. PMID- 16255519 TI - [Initial evaluation of EBM sequelae. More points for family practitioners, break in by specialized internist]. PMID- 16255520 TI - [Benign prostatic syndrome. Urinary urgency and micturition frequency reduced with plant preparation]. PMID- 16255521 TI - [Fever after stay in the tropics]. AB - This article provides an overview of possible causes of fever after traveling in tropical and subtropical areas and emphasizes the necessity of a conclusive diagnosis. PMID- 16255522 TI - [Bitter end of long-term alcoholism. Tension to the end]. PMID- 16255523 TI - Workers' compensation benefits--beneficios de la compensacion del trabajador. PMID- 16255524 TI - What are some challenges related to return to work policies, procedures, and processes? PMID- 16255525 TI - Innovative workplace influenza program: boosting employee immunization rates. AB - Planning and implementing a successful immunization program at the worksite requires a committed, interdepartmental effort and a dedicated occupational health staff. Planning should start well in advance of influenza season with adherence to strict timelines. Key strategies for campaign success include generating a sense of enthusiasm about the campaign; creating an employee- and family-friendly atmosphere to diffuse possible uneasiness about receiving vaccinations; assuring easy access to vaccines; and providing employees with evidence-based information related to influenza and immunization using a variety of media. Using the proven best practices of its on-site employee health vendor and adapting other strategies, the vaccine manufacturer succeeded in raising employee influenza immunization rates from 34% in 2002 to 66% by the end of November 2003, well beyond the program's goal of 50%. By the end of December, after extra clinics were added in response to intense media coverage, the employee immunization rate increased to 76%. PMID- 16255526 TI - Proper positioning of clients: a risk for caregivers. AB - The primary objective of this study was to learn more about the occupational risk of client repositioning by caregivers. This study also included information on frequency demands and factors contributing to the need to reposition clients and future risk-reduction solutions and interventions. A questionnaire was administered to caregivers at 12 hospitals. Targeted groups were client care units with caregivers who are required to reposition clients in bed. The task of client repositioning presents a high frequency demand to caregivers. Some characteristics such as size and health condition made it more likely clients would require repositioning. Researchers need to study client repositioning further and look for methods that will reduce the caregiver occupational risk factors. Caregivers are seeking new devices and techniques that facilitate client repositioning and benefit both the caregiver and the client. PMID- 16255527 TI - Employee perceptions of diabetes education needs: a focus group study. AB - The purpose of this descriptive non-experimental research was to assess employee perceptions of desired diabetes education through focus groups. Thirteen employees of a southern university in three focus groups identified many standard and some emerging educational topics of interest including healthier food choices when eating out, increasing activity, deciding whether highly advertised "special" products for diabetes were necessary, and recognizing the importance of obesity among children. The employees were willing to attend group meetings related to diabetes, but not to pay for them. The information will be used to plan an intervention to promote diabetes prevention and self-management in a worksite environment where diabetes costs are rising rapidly, and to support the need for a university wellness program. Occupational health nurses can address both employees' needs and employers' costs through such a program. PMID- 16255528 TI - Ergonomic evaluation: part of a treatment protocol for musculoskeletal injuries. AB - Ergonomic analyses and interventions are used as primary prevention methods to reduce physical stressors in the workplace and to prevent work-related musculoskeletal disorders (WMSDs). These methods can also be used for the treatment of injured employees. In this study, 103 employees with WMSDs resulting in more than 5 days away from usual work received an ergonomic evaluation which consisted of observation of usual work tasks, recommendations to minimize identified stressors, and case coordination. The goal of the intervention was to make simple job changes that would assist employees to return safely to usual job duties. The process for implementing this protocol for health care, airline, and university employees is described. The results show that after ergonomic evaluations were performed, the majority of recommendations were fully or partially (89%) implemented. Behavior changes were more likely to occur than administrative and equipment changes (p < .001). Occupational health nurses can use a similar program to enhance treatment plans for clients with WMSDs. PMID- 16255529 TI - Understanding cholesterol. PMID- 16255530 TI - [Recommendations of the 5th meeting of the Russian Gastroenterologists Scientific Society regarding cholelithiasis]. PMID- 16255531 TI - [Current views of vascular endothelium in health and at a pathology of the gastrointestinal tract]. AB - The article presents current data on the structure, function and biological role of vascular endothelium as well as its role in the genesis of pathologies of the gastrointestinal tract. Characteristics of endothelial vasoconstrictors and vasodilators are given. When we take into account the literature data and authors' studies, it becomes expedient to conduct further studies of the role of endothelial dysfunction in the development of diseases of the digestive tract in view of little knowledge of it at this pathology as well as availability of mostly experimental data. PMID- 16255533 TI - [Continuum of Helicobacter pylori infection as the scientific substantiation for taking clinical decisions]. AB - The article describes diseases of the gastroduodenal zone in the continuum of Helicobacter pylori infection. It discloses factors of risk and state of human health that increase the chance of the development of this or that disease and have an effect on its development and its complications that may lead to death. The article also disclosed a possibility of arresting the man's movement along the way of this continuum. PMID- 16255532 TI - [Neurogenic and genetic reasons and mechanisms of ulcers]. PMID- 16255534 TI - [Yeast-like fungi in the gastric mucus at acid-dependent disease]. PMID- 16255535 TI - [Juxtapapillary duodenal diverticula and pancreatitis]. PMID- 16255536 TI - [New alternative drug therapy for reducing symptoms associated with the functional gastric disorders]. PMID- 16255537 TI - [Correction of the motor-evacuation function of the postoperative stomach in the early postoperative period]. AB - We conducted the analysis of the results of surgical treatment of 255 patients with duodenal ulcers. In the early postoperative period the evacuation and motility function of the postoperative stomach was corrected with the use of the autonomous electrostimulator of the gastrointestinal tract (EAS GT) in all patients - in the main group (n = 125) and drug stimulation (cerucal 2.0 intramuscularly twice a day) and in the control group (n = 130). The use of EAS GT-3 certainly reduces symptoms of dyscoordination of the motor function of the gastrointestinal tract. PMID- 16255538 TI - [Application of videolaparoscopy methods in the diagnostics and treatment of acute appendicitis]. AB - The article describes an experience of the diagnostics and treatment of 550 patients who were hospitalized with acute appendicitis. Diagnostic laparoscopy was performed for 200 patients with acute appendicitis as a presumptive diagnosis. 350 patients underwent laparoscopic appendectomy. It was possible to avoid ineffective laparotomy in the course of diagnostic laparoscopy in 88% of patients with unclear symptoms of acute appendicitis, and to establish other acute diseases of the abdominal cavity in 12% of cases. Laparoscopic appendectomy can be performed practically at any form of appendix inflammation. It is better to carry out laparoscopic appendectomy using the clip method, and to treat the mesoappendix by monocoagulation. PMID- 16255539 TI - [Particular features of the clinical picture and nature of functional disorders in patients suffering from rectal prolapse]. PMID- 16255540 TI - [PPI-test as a differential criterion of selection for gastroesophageal reflux disease surgery]. PMID- 16255541 TI - [Study of proliferation and apoptosis parameters of gastric epithelium cells in patients with precancerous pathologies of the stomach]. AB - Changes in proliferation and apoptosis parameters of the gastric epithelium in patients with chronic Hp-associated gastritis were studied depending on the prevalence of the inflammatory process, presence of intestinal metaplasia and atrophic changes of the stomach glandular epithelium. The obtained results demonstrated a change in the cellular updating processes of the stomach epithelium depending on the localization, prevalence of the process, and more expressed - during the development of atrophic and dystrophic changes of the stomach mucous coat. Moderate amplification of apoptosis was recorded in focal atrophic gastritis, which was shown in the increased Iapopt and reduction of IPCNA and IcyclineD1 parameters. From then on, along with the distribution of the process, proliferation enhancement, occurrences of intestinal metaplasia and neoplasia centers, oppression of apoptosis destructions of stomach epitheliocytes was marked. PMID- 16255542 TI - [Experimental study of the effect of the phytomixture made of leaves of Plantago major L. and Achillea millenfolium L. on the secretion activity of the stomach in dogs]. AB - In this study, the author used plants with an obvious anti-ulcerous effect, which are often jointly included in gastroenterological phytomixtures. It was shown that the extract from leaves of Plantago major L. has a stimulating effect on gastric secretion, mostly on parietal cells. The extract from Achillea millenfolium L. reduced aggressive properties and enhanced protective properties of gastric juice. The common effect was demonstrated by the increased acid production and enhanced protective properties of gastric mucus. PMID- 16255543 TI - [Chronic EBV-infection in children with a high level of anti-H+/K+-ATPhase antibodies in gastric parietal]. AB - We examined 50 children at the age of 4-17 suffering from chronic gastritis. AntiH+/K+-ATPhase antibodies of gastric parietal cells, anti-EBV antibodies IgG EA and IgM VCA were determined with the help of immune-enzyme analysis methods. The first group comprised 28 children with a high level of antiparietal antibodies in the blood serum, and 22 children with a normal level of antiparietal antibodies made up the second group. The HP identification was carried out for all children. Anti-EBV antibodies IgG EA were found in 72% of children, and there were no statistical differences in the frequency and level of antibodies IgG EA. IgM VCA were not found in any of the patients. However, HP infection was revealed in 64.3% in the first group vs. 22.7% in the second group (p < 0,05); the combination of HB and chronic EBV infection was also more frequent in the first group than in the second group (42.8% and 18.1%, respectively, p < 0,05). PMID- 16255545 TI - [Clinical and economic analysis of the application of ranitidine and famotidine for treatment of duodenal ulcers]. PMID- 16255544 TI - [Celiac disease in adults: current state of the problem and description of a rare clinical case]. PMID- 16255546 TI - [Diagnosis and therapy of acid-dependent diseases including those associated with Helicobacter pylori]. PMID- 16255547 TI - [Etiopathogenesis, diagnostics and treatment of osteoporosis at inflammatory bowel diseases]. AB - Osteoporosis and related osteoporotic bone fractures are more and more becoming causes of disability for many categories of patients leading to the reduction of their life quality as well as high direct and indirect material expenses. This is true in full measure for patients with inflammatory bowel diseases (IBD) who experience a negative effect produced on the mineral density of bone tissue by long-term chronic inflammation, impaired calcium and vitamin D absorption, hypotrophy and, finally, therapy with glucocorticoids. This stimulates clinicians to develop actively the osteoporosis screening methods for patients with IBD as well as methods of its prevention and treatment. PMID- 16255548 TI - [Diseases associated with Helicobacter pylori and Candida spp.: clinical logic of combined studies]. AB - This article discusses problems of clinical value of one of the most widespread gastroenterological forms of candidal digestive apparatus lesions--candidal dysbiosis of the bowels representing an interdisciplinary problem in its essence being at the turn of gastroenterology and clinical mycology. The article also shows the clinical and pathogenetic need in combined studies of helicobacteriosis of the stomach and candidosis of the bowels. It substantiates the approaches to the rational anti-mycotic therapy in the complex program of treatment for this category of patients. PMID- 16255549 TI - [Role of Helicobacter pylori in different extragastroduodenal diseases]. PMID- 16255550 TI - [Peptic ulcers of the gastroduodenal zone in patients with scheduled cardiac operation: continuum of heterogeneous pathogenetic effects]. AB - The article is a review of different pathogenetic mechanisms (infectious, NSAID associated, stress-dependent and those stipulated by microcirculation disturbances in the gastroduodenal zone) of stomach and duodenum ulcers in patients with scheduled surgical intervention on the heart with the use of cardiopulmonary bypass. Some vexed questions related to treatment tactics of peptic ulcers in these patients are discussed. PMID- 16255551 TI - [Clinical efficacy of activia tvorozhnaya in patients with the irritated bowels syndrome with the prevalence of constipations]. PMID- 16255552 TI - [Impact of isovolumic hemodilution on the local hemodynamics of the resected stomach in patients with ulcer disease]. AB - The influence of isovolumic hemodilution on the local hemodynamics of the resected stomach in patients with a complicated course of duodenal ulcer was studied. In the course of the analysis of parameters of the local blood flow of the intact and resected stomach, the best indices were received in patients with preoperative isovolumic hemodilution. Preoperative isovolumic hemodilution in patients with a complicated course of duodenal ulcer after the stomach resection reduces the risk of early postoperative complications. PMID- 16255553 TI - [Express diagnostics, surgical tactics and efficacy of treating patients with diffuse peritonitis]. AB - The results of treatment of 100 patients with widespread forms of peritonitis were analyzed. In the control group of patients complications were diagnosed with the use of intraluminal pulsemotorography by Z.M. Sigal (1981) and assessment of the degree of endotoxicosis by the new original methodology. Intraperitoneal complications in the control group made up 8.0%, and extraperitoneal complications--2.0%. Intraperitoneal complications in the group of comparison made up 16.0%, and extraperitoneal complications--18.0%. PMID- 16255554 TI - [Selection of the surgery method for patients with high "stomach ulcers"]. PMID- 16255555 TI - [Selection of the method of treatment for patients with rectal prolapse]. PMID- 16255556 TI - [Proposal for a new endoscopic classification of the gastroesophageal reflux disease]. PMID- 16255557 TI - [Assessment of some metabolic parameters of white rats' liver after exposure to repetitive x-ray or microwave pulses]. AB - Effects of repetitive X-ray and microwave pulses on the rat liver functions were investigated. The action of repetitive nanosecond X-ray is characterized by the metabolic dysfunction of the liver. In particular, it results in a considerable reduction in the ALT activity, augmentation of the AST/ALT ratio and decrease of the total protein content. The most considerable effect is observed at 16 Hz. Microwave pulses render a less significant effect on metabolic functions of the rat liver as compared to X-rays. The effect depends on the frequency of pulses. PMID- 16255558 TI - [Efficacy of the new collection of herbs at stressful experimental sharp ulcer defects of the gastroduodenal zone]. AB - As a result of experiments on white male rats after an immobilization stress that resulted in the formation of sharp ulcer defects in the gastroduodenal zone, it was established that the daily 5-day peroral administration of the collection of herbs (Korniozil) in the volume of 4-6 ml in equal dozes 3 times a day as 9-12% of a water-alcohol solution had a positive influence on the regeneration of the mucous coat of the stomach. The metabolism and structure of lipids in the mucous coat of the stomach as well as balance between activity of processes of peroxide lipid oxidations and functioning of antioxidant systems was simultaneously restored. PMID- 16255561 TI - Health savings accounts will solve Medicare problem. PMID- 16255562 TI - Patients want a friendly face. PMID- 16255559 TI - [Functional state of the hematosalivary barrier in children with gastroduodenal diseases]. AB - The hematosalivary barrier state was studied using the values of peroxidation of lipids and nitric oxide, antioxidant defense and macroelements in children with chronic gastroduodenitis and functional dyspepsia. The disturbances of the barrier functioning were shown to be important in the mechanisms of stomach and duodenum lesions. Taking into account significant changes in the metabolic profile of the sputum, which are different for different inflammatory and functional diseases of the gastroduodenal zone, we have suggested using its parameters for noninvasive screening diagnostics of this pathology. PMID- 16255563 TI - Union needed. PMID- 16255564 TI - Overheard in the doctors lounge. PMID- 16255565 TI - A steady pace to equality. PMID- 16255566 TI - Women doctors. PMID- 16255567 TI - Quality or cost control? PMID- 16255568 TI - First line of defense. PMID- 16255569 TI - Apples and oranges. PMID- 16255570 TI - The EHR payoff. PMID- 16255571 TI - Desorption electrospray ionization of explosives on surfaces: sensitivity and selectivity enhancement by reactive desorption electrospray ionization. AB - Desorption electrospray ionization (DESI), an ambient mass spectrometry technique, is used for trace detection of the explosives trinitrohexahydro-1,3,5 triazine (RDX), octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (HMX), 2,4,6 trinitrotoluene (TNT), Pentaerythritol tetranitrate (PETN), and their plastic compositions (Composition C-4, Semtex-H, Detasheet) directly from a wide variety of surfaces (metal, plastic, paper, polymer) without sample preparation or pretreatment. Analysis of the explosives is performed under ambient conditions from virtually any surface in very short times (<5 s) including confirmatory tandem mass spectrometry (MS/MS) experiments, while retaining the sensitivity and specificity that mass spectrometry offers. Increased selectivity is obtained both by MS/MS and by performing additional experiments in which additives are included in the spray solvent. These reactive DESI experiments (reactions accompanying desorption) produce such ions as the chloride and trifluoroacetate adducts of RDX and HMX or the Meisenheimer complex of TNT. Desorption atmospheric pressure chemical ionization, a variant of DESI that uses gas-phase ions generated by atmospheric pressure corona discharges of toluene or other organic compounds, provides evidence for a heterogeneous-phase (gaseous ion/absorbed analyte) charge transfer mechanism of DESI ionization in the case of explosives. Plastic explosives on surfaces were analyzed directly as fingerprints, without sample preparation, to test DESI as a possible method for in situ detection of explosives-contaminated surfaces. DESI also allowed detection of explosives in complex matrixes, including lubricants, household cleaners, vinegar, and diesel fuel. Absolute limits of detection for the neat explosives were subnanogram in all cases and subpicogram in the case of TNT. The DESI response was linear over 3 orders of magnitude for TNT. Quantification of RDX on paper gave a precision (RSD) of 2.3%. Pure water could be used as the spray solution for DESI, and it showed ionization efficiencies for RDX in the negative ion mode similar to that given by methanol/water. DESI represents a simple and rapid way to detect explosives in situ with high sensitivity and specificity and is especially useful when they are present in complex mixtures or in trace amounts on ordinary environmental surfaces. PMID- 16255572 TI - Identifying the SPARC binding sites on collagen I and procollagen I by atomic force microscopy. AB - SPARC (secreted protein acidic and rich in cysteine) is a matricellular protein associated with the extracellular matrix (ECM). It has been found that the production of collagen I is a requisite for the association of SPARC with ECM, and studies with SPARC-null mice indicate that SPARC plays a role in modifying the structure of collagen fibers. It is not known, however, whether SPARC interacts with the collagen I precursor, procollagen I. In this study, the binding of SPARC to collagen I and procollagen I was verified by surface plasmon resonance. The SPARC-binding sites on collagen I and procollagen I were identified by directly visualizing their complexes using tapping-mode atomic force microscopy (TM-AFM). The characteristic chain end feature in collagen I is not readily detected by AFM, so unambiguous location of the binding sites relative to the C- or N-termini is difficult. In contrast, procollagen I, with its large globular C-propeptide, permits easy identification of the C-terminus. Histograms were constructed and compared based on the distances of the bound SPARC to the C-terminus of procollagen I and to the closest end of collagen I. There is a broad distribution of SPARC binding sites on procollagen I with the most preferred binding region located approximately 1/3 from the C-terminus. Characterization of the SPARC-binding sites on collagen I and procollagen I provides useful information for further understanding of the functional implications of their interactions. PMID- 16255573 TI - Electrokinetic transport in nanochannels. 1. Theory. AB - Electrokinetic transport in fluidic channels facilitates control and separation of ionic species. In nanometer-scale electrokinetic systems, the electric double layer thickness is comparable to characteristic channel dimensions, and this results in nonuniform velocity profiles and strong electric fields transverse to the flow. In such channels, streamwise and transverse electromigration fluxes contribute to the separation and dispersion of analyte ions. In this paper, we report on analytical and numerical models for nanochannel electrophoretic transport and separation of neutral and charged analytes. We present continuum based theoretical studies in nanoscale channels with characteristic depths on the order of the Debye length. Our model yields analytical expressions for electroosmotic flow, species transport velocity, streamwise-transverse concentration field distribution, and ratio of apparent electrophoretic mobility for a nanochannel to (standard) ion mobility. The model demonstrates that the effective mobility governing electrophoretic transport of charged species in nanochannels depends not only on electrolyte mobility values but also on zeta potential, ion valence, and background electrolyte concentration. We also present a method we term electrokinetic separation by ion valence (EKSIV) whereby both ion valence and ion mobility may be determined independently from a comparison of micro- and nanoscale transport measurements. In the second of this two-paper series, we present experimental validation of our models. PMID- 16255574 TI - Electrokinetic transport in nanochannels. 2. Experiments. AB - We present an experimental study of nanoscale electrokinetic transport in custom fabricated quartz nanochannels using quantitative epifluorescence imaging and current monitoring techniques. One aim is to yield insight into electrical double layer physics and study the applicability of continuum theory to nanoscale electrokinetic systems. A second aim is to explore a new separation modality offered by nanoscale electrophoretic separations. We perform parametric variations of applied electric field, channel depth, background buffer concentration, and species valence to impose variations on zeta potential, effective mobility, and Debye length among other parameters. These measurements were used to validate a continuum theory-based analytical model presented in the first of this two-paper series. Our results confirm the usefulness of continuum theory in predicting electrokinetic transport and electrophoretic separations in nanochannels. Our model leverages independent measurements of zeta potential performed in a microchannel system at electrolyte concentrations of interest. These data yield a zeta potential versus concentration relation that is used as a boundary condition for the nanochannel electrokinetic transport model. The data and model comparisons together show that the effective mobility governing electrophoretic transport of charged species in nanochannels depends not only on ion mobility values but also on the shape of the electric double layer and analyte ion valence. We demonstrate a method we term electrokinetic separation by ion valence, whereby both ion valence and mobility may be determined independently from a comparison of micro- and nanoscale transport measurements. PMID- 16255575 TI - Fabrication and analysis of spatially uniform field electrokinetic flow devices: theory and experiment. AB - A uniform-field design approach can improve the performance of microanalytical, chip-based devices for a number of applications, including separations and sample preparation. The faceted prism paradigm allows the design of microfluidic devices possessing spatially uniform fields in electrokinetically driven flows. We present the first quantitative study of the velocity fields obtained using faceted interfaces between deep and shallow channel sections. Electrokinetic flows were generated in a series of wet-etch fabricated microfluidic channels. The resulting velocity fields were analyzed by particle image velocimetry and compared with simulations of the two-dimensional Laplace equation using both the designed channel geometry and the as-fabricated channel geometry. This analysis found localized differences between the designed and observed flow fields that were directly attributable to the limitations of isotropic substrate etching. Simulations using the as-fabricated channel geometry reproduced the experimental electrokinetic velocity field, quantitatively accounting for speed field variations due to the limits of the fabrication method. The electrokinetic speed fields were also compared to corresponding pressure-driven speed fields. PMID- 16255576 TI - Dielectrophoretic manipulation of particles and cells using insulating ridges in faceted prism microchannels. AB - This paper presents a novel device for the dielectrophoretic manipulation of particles and cells. A two-level isotropic etch of a glass substrate was used to create three-dimensional ridge-like structures in micrometer-sized channels. Due to the insulating properties of glass, locally patterned regions of nonuniform electric field form near the ridges when a dc field is applied along the channel. The ridges are designed using the method of faceted prisms, such that substantially uniform fields are produced on each side of the faceted interfaces that form each ridge. The dielectrophoretic force that results from the electric field gradient near the ridges is used to affect particle motion parallel to the ridges in the absence of a bulk pressure-driven flow. Trapping and deflection of particles and continuous concentration and separation of Bacillus subtilis from a two-component sample mixture are demonstrated. The flow of B. subtilis is restricted to a selected channel of a planar, multichannel device as a result of negative dielectrophoresis arising from the presence of the insulating ridges when the applied electric field exceeds a threshold of 30 V/mm. Dielectrophoresis has a negligible impact on 200-nm-diameter polystyrene particles under the same conditions. PMID- 16255577 TI - X-Hitting: an algorithm for novelty detection and dereplication by UV spectra of complex mixtures of natural products. AB - A major challenge in lead discovery is to detect well-known and trivial compounds rapidly, a process known as dereplication, so that isolation, structure elucidation, and pharmacological investigations can be focused on novel compounds. In this paper, we present a new algorithm, X-hitting, based on cross sample comparison of full UV spectra from HPLC analysis of highly complex natural product extracts/samples. X-Hitting allows automatic identification of known compounds but more important also allows finding of potentially new or similar compounds. We demonstrate this new algorithm by automatic identification of known structures, a task we call cross-hitting, and tentative identification of potentially new bioactive compounds, a task we call new-hitting, in HPLC data from analysis of fungal extracts. Both tasks are illustrated using 18 important reference compounds and complex fungal extracts obtained from isolates in the IBT Culture Collection held at BioCentrum-DTU, Technical University of Denmark. The receiver operating characteristics statistic is used to evaluate the performance of the compound predictor, and it was found that compounds could be identified with high confidence (AUC approximately 0.98). Based on high confidence in retrieving identical spectra, the method is extended to include similar but still different spectra. PMID- 16255578 TI - Multidimensional high-throughput screening of displacers. AB - A multidimensional, batch high-throughput screening (MD-HTS) protocol was developed to investigate the effects of various parameters on the selectivity of ion-exchange protein displacement systems. A variety of molecules were screened, and the results were employed to provided insights into the influence of displacer chemistry and concentration, resin chemistry, and mobile-phase salt counterion on the efficacy and selectivity of these nonlinear chromatographic systems. These results open up the possibility of tailoring the selectivity of displacement separations by choosing appropriate combinations of operating conditions using the MD-HTS technique. The screens were also employed for the identification of displacers and conditions for the separation of a challenging protein mixture by selective displacement chromatography. Column displacements were carried out with potential lead compounds identified from the MD-HTS screens, and the results confirmed that selective displacement could indeed be achieved for this model mixture. Furthermore, the results indicated that this approach is particularly useful when the order of elution is not changed, but the inherent selectivity is increased in the presence of the displacer. The results presented in this paper demonstrate the utility of the MD-HTS technique for rapid method development in protein ion-exchange displacement chromatography. PMID- 16255579 TI - Encapsulation of enzymes within polymer spheres to create optical nanosensors for oxidative stress. AB - We describe the fabrication and characterization of poly(ethylene glycol) (PEG) hydrogel spheres containing the enzyme horseradish peroxidase (HRP) for application as optical nanosensors for hydrogen peroxide. HRP was encapsulated in PEG hydrogel spheres by reverse emulsion photopolymerization, yielding spheres with a size range from 250 to 350 nm. Encapsulated HRP activity and sensitivity to hydrogen peroxide were investigated by the Amplex Red assay based on the fluorescence response as a function of H2O2. These HRP-loaded spheres were then introduced to murine macrophages with Amplex Red in the culture media. After phagocytosis, the biocompatibility of spheres was determined by live cell staining using calcein AM (5 microM). The HRP-loaded PEG hydrogel spheres were activated (i.e., fluorescent oxidized Amplex Red produced within the spheres) by oxidative stresses such as exogenous H2O2 (100 microM) and lipopolysaccharide (1 microg/mL), which induced the production of endogenous peroxide inside macrophages. The results presented here indicate that after polymerization, the enzyme activity of HRP was still maintained and that using these HRP-containing nanospheres, peroxide production could be sensed locally within cells. PMID- 16255580 TI - Engineered recombinant single-chain fragment variable antibody for immunosensors. AB - A recombinant single-chain fragment variable (scFv) antibody (designated A10B) was engineered to contain two histidines within the linker peptide used to join the scFv heavy and light chains. A piezoimmunosensor using the scFv was successfully developed. A10B scFv bound to the gold piezoimmunosensor surface were correctly oriented, retained antigen-binding activity, and coupled at high surface concentration. These results, and results obtained from an earlier study using an scFv containing a linker cysteine, suggest that the location on the linker sequence in which the amino acids were incorporated was well tolerated by the scFv and did not interfere with scFv antigen-binding activity. The scFv modified QCM sensor was thoroughly characterized and used to specifically detect antigen in crude serum sample and had a sensitivity of 2.3 +/- 0.15 nM (n = 4) with a linear range over 2.3 x 10(-9) - 3.3 x 10(-(8) M. The piezoimmunosensor was also used to study the kinetics and thermodynamics of antigen/scFv antibody binding. PMID- 16255581 TI - Microfluidic biosensor based on an array of hydrogel-entrapped enzymes. AB - Here we show that a microfluidic sensor based on an array of hydrogel-entrapped enzymes can be used to simultaneously detect different concentrations of the same analyte (glucose) or multiple analytes (glucose and galactose) in real time. The concentration of paraoxon, an acetylcholine esterase inhibitor, can be quantified using the same approach. The hydrogel micropatch arrays and the microfluidic systems are easy to fabricate, and the hydrogels provide a convenient, biocompatible matrix for the enzymes. Isolation of the micropatches within different microfluidic channels eliminates the possibility of cross talk between enzymes. PMID- 16255582 TI - Chip-based on-line nanospray MS method enabling study of the kinetics of isocyanate derivatization reactions. AB - The reaction of propyl isocyanate (2), benzyl isocyanate (3), and toluene-2,4 diisocyanate (4) with 4-nitro-7-piperazino-2,1,3-benzoxadiazole (1) to yield the corresponding urea derivatives 5 was carried out in a continuous flow glass microfluidics chip. Real-time monitoring of the derivatization reactions was done by electrospray ionization mass spectrometry, making use of a recently reported modular chip-MS interface. Rate constants of 1.5 x 10(4), 5.2 x 10(4), and 2.4 x 10(4) M(-1) min(-1) were determined for 2, 3, and 4, respectively. Using macroscale batch conditions, the rate constants are 3-4 times lower. The faster on-chip kinetics is attributed to the more efficient molecular diffusion in the micrometer-sized channel. PMID- 16255583 TI - On-chip microfluidic transport and mixing using electrowetting and incorporation of sensing functions. AB - An integrated system was developed that performs microfluidic transport, mixing, and sensing on a single chip. The operation principle for the microfluidic transport was based on electrowetting. A solution to be transported was confined in a space between a row of gold working electrodes and a protruding poly(dimethylsiloxane) (PDMS) structure. When a negative potential was applied to one of the gold working electrodes, it became hydrophilic, and the solution was transported through the flow channel. The solution could be transported in any desired direction in a network of flow channels by switching on necessary electrodes one by one. Furthermore, two solutions transported through two flow channels could be mixed using a mixing electrode based on the same principle. To demonstrate the applicability of a lab-on-a-chip, an air gap ammonia electrode was integrated by taking advantage of the open structure of the flow channel. Gaseous ammonia that was produced after pH adjustment and diffused through an air gap caused a pH change in the electrolyte layer, which was measured with an iridium oxide pH indicator electrode. The 90% response time was less than 1 min for the millimolar order of ammonia. The calibration curve was linear down to 10 microM. The ammonia-sensing system was also applied to construct biosensing systems for urea and creatinine. A linear relationship was observed between the potential and the logarithm of the concentration of the analytes down to 50 microM for both urea and creatinine. The developed microfluidic system can be a basic building block for future systems. PMID- 16255584 TI - Microarray immunoassay for phenoxybenzoic acid using polymer encapsulated Eu:Gd2O3 nanoparticles as fluorescent labels. AB - Currently, detection in microarray bioanalysis is based mainly on the use of organic dyes. To overcome photobleaching and spectral overlaps we applied a new type of fluorophore, crystalline europium-doped gadolinium oxide (Eu:Gd2O3) nanoparticles, as labels in immunoassay microarrays. The Eu:Gd2O3 nanoparticles synthesized by spray pyrolysis offer narrow red emission, large Stokes shift, photostable laser-induced fluorescence with a long lifetime (1 ms). The amino functionalization of the particles was achieved by poly(L-lysine) (PL) encapsulation. The formation of a stable PL shell was confirmed by TEM analysis, colloidal stability studies, and quantification of the surface reactive amino groups. The PL-encapsulated particles were covalently conjugated to antibodies and successfully applied as reporters in a competitive fluorescence microimmunoassay for phenoxybenzoic acid (PBA), a generic biomarker of human exposure to pyrethroid insecticides. Microarrays were fabricated by microcontact printing of BSA-PBA in line patterns (10 x 10 microm). Confocal fluorescence microscopy combined with internal standard (fluorescein) calibration was used for quantitative measurements. The microarray immunoassay demonstrated a limit of detection of 1.4 microg L(-1) PBA. This work suggests the potential application of lanthanide oxide nanoparticles as fluorescent probes in microarray and biosensor technology, immunodiagnostics, and high-throughput screening. PMID- 16255585 TI - Capillary electrochromatography-mass spectrometry of zwitterionic surfactants. AB - This work describes the on-line hyphenation of a packed capillary electrochromatography (CEC) column with an internally tapered tip coupled to electrospray ionization-mass spectrometry (ESI-MS) and atmospheric pressure chemical ionization-mass spectrometry (APCI-MS) for the analysis of betaine-type amphoteric or zwitterionic surfactants (Zwittergent). A systematic investigation of the CEC separation and MS detection parameters comparing ESI and APCI is shown. First, a detailed and optimized manufacturing procedure for fabrication of the CEC-MS column with a reproducible internally tapered tip (7-9 microm) is presented. Next, the optimization of the separation parameters by varying the C(18) stationary-phase particle size (3 versus 1.5 microm), as well as mobile phase composition including acetonitrile (ACN) volume fraction, ionic strength, and pH is described. The optimized separation is achieved using 3-microm C(18) packing with 75% ACN (v/v), 5 mM Tris at pH 8.0. Optimization for on-line CEC-ESI MS detection is then done varying both the sheath liquid and spray chamber parameters while evaluating the use of random versus structured factorial table experimental designs. The more structured approach allows fundamental analysis of individual ESI-MS parameters while minimizing CEC and MS equilibration time between settings. A comparison of CEC-ESI-MS to CEC-APCI-MS using similar sheath and spray chamber conditions presents new insight for coupling of CEC to APCI-MS. The sheath liquid flow rate required to maintain adequate sensitivity is much higher in APCI source (50 microL/min) as compared to the ESI source (3 microL/min). The on-line mass spectra obtained in the full scan mode show that fragmentation in the two sources occurs at different positions on the Zwittergent molecules. For ESI-MS, the protonated molecular ion is always highest in abundance with minor fragmentation occurring due to the loss of the alkyl chain. In contrast, the APCI-MS spectra show that the highest abundant ion resulted by elimination of propane sulfonate from the Zwittergent molecule. A comparison of the sensitivity between the two sources in positive ionization SIM mode shows that CEC-ESI-MS provides an impressive limit of detection (LOD) of 5 ng/mL, which is at least 3 orders of magnitude lower than CEC-APCI-MS (LOD 100 microg/mL). Finally, the optimized CEC-MS methods comparing ESI and APCI are applied for separation and structural characterization of a real industrial zwittergent sample, Rewoteric AM CAS. PMID- 16255586 TI - Ultrarapid desalting of protein solutions for electrospray mass spectrometry in a microchannel laminar flow device. AB - The adverse effects of nonvolatile salts on the electrospray (ESI) mass spectra of proteins and other biological analytes are a major obstacle for a wide range of applications. Numerous sample cleanup approaches have been devised to facilitate ESI-MS analyses. Recently developed microdialysis techniques can shorten desalting times down to several minutes, the bottleneck being diffusion of the contaminant through a semipermeable membrane. This work introduces an approach that allows the on-line desalting of macromolecule solutions within tens of milliseconds. The device does not employ a membrane; instead, it uses a two layered laminar flow geometry that exploits the differential diffusion of macromolecular analytes and low molecular weight contaminants. To maximize desalting efficiency, diffusive exchange between the flow layers is permitted only for such a time as to allow full exchange of salt, while incurring minimal macromolecule exchange. Computer simulations and optical studies show that the device can reduce the salt concentration by roughly 1 order of magnitude, while retaining approximately 70% of the original protein concentration. Application of this approach to the on-line purification of salt-contaminated protein solutions in ESI-MS results in dramatic improvements of both the signal-to-noise ratio and the absolute signal intensity. However, efficient desalting requires the diffusion coefficients of salt and analyte to differ by roughly 1 order of magnitude or more. This technique has potential to facilitate high-throughput analyses of biological macromolecules directly from complex matrixes. In addition, it may become a valuable tool for process monitoring and for on-line kinetic studies on biological systems. PMID- 16255587 TI - Channel flow configuration for studying the kinetics of surfactant polyelectrolyte binding. AB - A novel channel flow configuration was developed and utilized for studying the poly(styrenesulfonate)-cetylpyridinium ion interaction kinetics. The surfactant solution was continuously injected into a flow of polyelectrolyte solution, and the extent of the association reaction was probed at an ion-selective detector electrode. The system was modeled within an analytical approximation, which was tested by a finite-element simulation of the full convective mass transport problem including the homogeneous complexation reaction. The results show that association kinetics can be resolved and that the initial steps of the reaction are not influenced by intermolecular interactions between the bound surfactants. The presented methodology is general, and further development should enable the study of complex cooperative kinetics of surfactant-polyelectrolyte systems. PMID- 16255588 TI - Analysis of biogenic amine variability among individual fly heads with micellar electrokinetic capillary chromatography-electrochemical detection. AB - Neurochemical variability among individual Drosophila heads has been examined with the sensitivity of electrochemical detection and the selectivity of micellar electrokinetic capillary chromatography. Homogenization of single Drosophila heads in volumes as small as 100 nL has been accomplished. Here we demonstrate reproducible separations for single fly heads in 250-nL volumes providing a 4 fold increase in sensitivity without overloading the electrochemical detector. This increase in sensitivity allows detection of previously undetected analytes, such as N-acetyltyramine (naTA) and octopamine (OA). Analytes including L-3,4 dihydroxyphenylalanine, N-acetyl octopamine, N-acetyldopamine, naTA, N acetylserotonin, OA, dopamine, tyramine, and serotonin also have been consistently identified in single-head homogenates and observed with homogenates representing populations of Drosophila. Neurochemical variation between individual flies as well as the consistency within a population indicates varying amounts of neurotransmitter turnover. The inception, design, and fabrication of a miniature tissue homogenizer has enabled the separation of biogenic amines and metabolites from these severely volume-limited single Drosophila head homogenates. PMID- 16255589 TI - Fabrication of discontinuous surface patterns within microfluidic channels using photodefinable vapor-based polymer coatings. AB - In this report, we introduce a surface modification method for the fabrication of discontinuous surface patterns within microfluidic systems. The method is based on chemical vapor deposition (CVD) of a photodefinable coating, poly(4-benzoyl-p xylylene-co-p-xylylene), onto the luminal surface of a microfluidic device followed by a photopatterning step to initiate spatially controlled surface binding. During photopatterning, light-reactive groups of the CVD polymer spontaneously react with molecules adjunct to the surface, such as poly(ethylene oxide). We demonstrate the potential of these reactive polymers for surface modification by preventing nonspecific protein adsorption on different substrates including silicon and poly(dimethylsiloxane) as measured by fluorescence microscopy. More importantly, three-dimensional patterns have successfully been created within polymer-based microfluidic channels, establishing spatially controlled, bioinert surfaces. The herein reported surface modification method addresses a critical challenge with respect to surface engineering of microfluidic devices, namely, the fabrication of discontinuous patterns within microchannels. PMID- 16255590 TI - Desorption electrospray ionization mass spectrometry for high-throughput analysis of pharmaceutical samples in the ambient environment. AB - Desorption electrospray ionization (DESI) allows mass spectrometry to be used for on-line high-throughput monitoring of pharmaceutical samples in the ambient environment, without prior sample preparation. Positive and negative ion DESI are used to characterize the active ingredients in pharmaceutical samples formulated as tablets, ointments, and liquids. Compounds of a wide variety of chemical types are detected in these complex matrices. The effects on analytical performance of operating parameters, including the electrospray high voltage, heated capillary temperature, solvent infusion rate, and solvent composition, are evaluated and optimized. In addition to experiments in which a simple solvent is sprayed onto the solid analyte samples, reactive desorption is performed by adding reagents to the solvent spray to generate particularly stable or characteristic ions with the analytes of interest. A variable-speed moving belt was built for high-throughput sampling and used to provide rapid qualitative and semiquantitative information on drug constituents in tablets. Sampling rates as high as 3 samples/s are achieved in the ambient environment. Relative standard deviations of the relative ion abundances for major components in the mass spectra are in the range of 2-8%. Impurities and components present at levels as low as approximately 0.1% are identified and carryover effects are minimized in high-throughput on-line analysis of pharmaceutical samples. PMID- 16255592 TI - Microfluidic device for electric field-driven single-cell capture and activation. AB - A microchip that performs directed capture and chemical activation of surface modified single cells has been developed. The cell capture system is comprised of interdigitated gold electrodes microfabricated on a glass substrate within PDMS channels. The cell surface is labeled with thiol functional groups using endogenous RGD receptors, and adhesion to exposed gold pads on the electrodes is directed by applying a driving electric potential. Multiple cell types can thus be sequentially and selectively captured on desired electrodes. Single-cell capture efficiency is optimized by varying the duration of field application. Maximum single-cell capture is attained for the 10-min trial, with 63 +/- 9% (n = 30) of the electrode pad rows having a single cell. In activation studies, single M1WT3 CHO cells loaded with the calcium-sensitive dye fluo-4 AM were captured; exposure to the muscarinic agonist carbachol increased the fluorescence to 220 +/ 74% (n = 79) of the original intensity. These results demonstrate the ability to direct the adhesion of selected living single cells on electrodes in a microfluidic device and to analyze their response to chemical stimuli. PMID- 16255591 TI - Quantitative determination of protein nuclear transport induced by phosphorylation or by proteolysis. AB - Nucleocytoplasmic transport of proteins in eukaryotic cells is a fundamental process for gene expression. The transport is regulated by posttranslational modifications of the proteins, such as ligand-binding, phosphorylation, and proteolysis. For monitoring the nuclear transport of proteins induced by a ligand binding, we have recently developed a genetically encoded bioluminescent indicator based on reconstitution of split fragments of Renilla reniformis (RLuc) by protein splicing with DnaE inteins. We herein describe that the technique is used for detecting phosphorylation- or proteolysis-induced nuclear transports of a target protein. Two model proteins, signal transducer and activator of transcription 3 (STAT3) and sterol-regulatory element binding protein-2 (SREBP 2), were exemplified as phosphorylation- and proteolysis-induced nuclear transport, respectively. Each STAT3 or SREBP-2 is connected with C-terminal halves of RLuc and DnaE. If the protein translocates into the nucleus, the C terminal fragment of RLuc meets the N-terminal fragment of RLuc, and full-length RLuc is reconstituted by protein splicing in the nucleus. The indicator with SREBP-2 enabled us to quantify the intracellular concentrations of cholesterol. The indicator with STAT3 quantified the extent of the nuclear transport induced by representative cytokines. This simple assay based on protein nuclear transports allows the selection of suitable drugs among candidates and has significant potential for risk assessments, such as carcinogenic chemical screening in vitro and in vivo. PMID- 16255593 TI - Optical determination of differential coverage-potential relations of redox active species immobilized on electrode surfaces. AB - Single-wavelength (HeNe laser, lambda = 633 nm), normal incidence UV-visible reflectance spectroscopy was used to monitor the optical properties of the glassy carbon (GC)|0.2 M NaOH(aq) interface as a function of the applied potential, E. Whereas the electroreflectance coefficient for bare GC was found to be small and potential independent, surface functionalization by an irreversibly adsorbed layer of tetrasulfonated cobalt phthalocyanine (CoTSPc) yielded a clearly defined sigmoidally shaped normalized reflectance change (DeltaR/R) vs E curve over the potential region in which the adsorbate displayed redox peaks. Assuming DeltaR/R is proportional to the extent of redox conversion, as has been reported for macrocycles adsorbed on other types of carbon (e.g., Kim, S.; Xu, X.; Bae, I. T.; Wang, Z.; Scherson, D. A. Anal. Chem. 1990, 62, 2647-2650), differential coverage potential relations were determined based purely on the optical data collected. A similar optical behavior was found for irreversible adsorbed CoPc and tetraamino CoPc (CoTAPc) adsorbed on GC, for which the voltammetric peaks were ill-defined, too small for coulometric analyses to be reliably performed, or both. No detectable changes in the DeltaR/R vs E profiles of either bare or macrocyclic functionalized surfaces were observed upon addition of hydrazine to the neat 0.2 M NaOH solution at potentials at which these surfaces display electrocatalytic properties for its oxidation. Possible factors responsible for this behavior are discussed. PMID- 16255594 TI - Microfluidic platform for liquid chromatography-tandem mass spectrometry analyses of complex peptide mixtures. AB - A microfluidic chip that integrates all the fluidic components of a gradient liquid chromatography (LC) system is described. These chips were batch-fabricated on a silicon wafer using photolithographic processes and with Parylene as the main structural material. The fabricated chip includes three electrolysis-based electrochemical pumps, one for loading the sample and the other two for delivering the solvent gradient; platinum electrodes for delivering current to the pumps and establishing the electrospray potential; a low-volume static mixer; a column packed with silica-based reversed-phase support; integrated frits for bead capture; and an electrospray nozzle. The fabricated structures were able to withstand pressures in excess of 250 psi. The device was used to perform a liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis of a mixture of peptides from the trypsin digestion of bovine serum albumen (BSA). Gradient elution through the 1.2-cm column was performed at a flow rate of 80 nL/min. Compared to the analysis of the same sample using a commercial nanoflow LC system, the chromatographic resolution was nearly as good, and the total cycle time was significantly reduced because of the minimal volume between the pumps and the column. Results demonstrate the potential of mass-produced, low-cost microfluidic systems capable of performing LC separations for proteomics applications. PMID- 16255595 TI - Negative-mode MALDI-TOF/TOF-MS of oligosaccharides labeled with 2-aminobenzamide. AB - MALDI-TOF-MS of 2-aminobenzamide-labeled N-glycans was shown to allow the analysis of sodium adducts and proton adducts in the positive-ion mode as well as deprotonated species in the negative-ion mode from a single preparation spot, using N-glycans of adult worms of the human parasite Schistosoma mansoni as model substances. Fragment ion analysis of these species was performed by MALDI-TOF/TOF MS. With laser-induced dissociation, sodium adducts and proton adducts mainly showed cleavage of glycosidic linkages. High-energy collision-induced dissociation of sodium adducts resulted in extensive cross-ring cleavages and provided information on linkage positions. Of particular value were the negative mode MALDI-TOF/TOF-MS analyses of the deprotonated N-glycans, which featured (1) various ring fragmentations giving linkage information, (2) extensive (1,3)A cross-ring cleavage of mannoses carrying an antenna readily revealing the composition of the antenna, (3) D as well as [D-18]- ions providing specifically the composition of the 6-antenna, and (4) pronounced stability of fucose linkages resulting in detailed information on fucosylation positions. The outlined approach thus allows the acquisition of both heCID MS/MS spectra of sodium adducts and LID MS/MS spectra of deprotonated species from a 2-aminobenzamide labeled N-glycan prepared in 6-aza-2-thiothymine, resulting in a wealth of structural information. PMID- 16255596 TI - Quantification of relative gene dosage by single-base extension and high performance liquid chromatography: application to the SMN1/SMN2 gene. AB - One of the most commonly used techniques for genotyping of single-nucleotide polymorphism (SNP) is detection of single-base extensions (SBEs). We present a new, rapid, simple, and highly reliable method for accurate quantification of SNP variants in a single reaction. Our approach is based on SBE detection coupled with high-performance liquid chromatography (HPLC) quantification. To demonstrate the utility of our approach, we report data to determine the gene dosage for relative amounts of alleles in a homologous gene, allowing detection of mutation causing exon skipping in human SMN genes to determine the ratio between the copy numbers of the SMN1/SMN2 gene. We successfully determined the relative ratio of the SMN1 and SMN2 genes and showed assay characteristics using the SBE reaction coupled with HPLC. This assay approach readily scaled to high parallelization with multiplex SBE reactions in a single sample screened in one analysis. By screening for particular SNP genotypes, this assay can be used to determine the relative gene dosage that correlates highly with the patient's disease state. The next challenge is to apply this novel methodology in a clinical screening and quantification setting for special gene regions within highly homologous genes. PMID- 16255597 TI - X-ray absorption fine structure combined with X-ray fluorescence spectrometry. Improvement of spectral resolution at the absorption edges of 9-29 keV. AB - X-ray absorption near-edge structure (XANES) suffers from core-hole lifetime broadening at a higher energy absorption edge, such as Sn K (29 keV, Gamma(K) = 8.49 eV). To overcome this problem, emitted Sn Kalpha1 fluorescence from sample was counted using high-energy-resolution fluorescence spectrometer in the XANES measurements. Experimental energy resolution (5.0 eV) was consistent with theoretical values based on the Rowland configuration of the spectrometer. The absorption edge became steeper compared to conventional spectra. The white-line peak due to Sn(II) species became remarkably sharper and more intense in the Sn Kalpha1-detecting Sn K-edge XANES for Pt-Sn/SiO2. To support the semiclassical theory of resonant Raman scattering for the explanation of observed elimination of lifetime width, more resolved XANES data at Cu K, Pb L3, and Sn K in this work were convoluted (filtered) with a Lorentzian of each core-hole lifetime width. The processed data resembled generally well corresponding XANES spectrum measured in transmission mode. The verification based on ab initio XANES calculations was also performed. PMID- 16255598 TI - Label-free detection of peptide nucleic acid-DNA hybridization using localized surface plasmon resonance based optical biosensor. AB - The development of label-free optical biosensors for DNA and other biomolecules has the potential to impact life sciences as well as screening in medical and environmental applications. In this report, we developed a localized surface plasmon resonance (LSPR) based label-free optical biosensor based on a gold capped nanoparticle layer substrate immobilized with peptide nucleic acids (PNAs). PNA probe was designed to recognize the target DNA related to tumor necrosis factor. The nanoparticle layer was formed on a gold-deposited glass substrate by the surface modified silica nanoparticles using silane-coupling reagent. The optical properties of gold-capped nanoparticle layer substrate were characterized through monitoring the changes in the absorbance strength, as the thickness of the biomolecular layer increased with hybridization. The detection of PNA-DNA hybridization with target oligonucleotides and PCR-amplified real samples were performed with a limit of detection value of 0.677 pM target DNA. Selective discrimination against a single-base mismatch was also achieved. Our LSPR-based biosensor with the gold-capped nanoparticle layer substrate is applicable to the design of biosensors for monitoring of the interaction of other biomolecules, such as proteins, whole cells, or receptors with a massively parallel detection capability in a highly miniaturized package. PMID- 16255599 TI - Colorimetric bio-barcode amplification assay for cytokines. AB - The bio-barcode amplification assay has become a powerful tool in detecting tens to hundreds of biological targets such as proteins and nucleic acids in the entire sample. However, current bio-barcode detection schemes still require many experimental steps including microarrayer-based immobilization of oligonucleotides on a glass chip, silver enhancement of immobilized gold nanoparticles on a chip, and light-scattering measurement. Here, we report a colorimetric bio-barcode method that minimizes the above requirements while detecting 30 aM concentrations of cytokines (approximately 3 orders of magnitude more sensitive than conventional nonenzymatic cytokine detection assays). The assay is based on porous microparticles, which enable loading of a large number of barcode DNA per particle, and gold nanoparticle-based colorimetric barcode detection method. PMID- 16255600 TI - Development of a cryogen-free concentration system for measurements of volatile organic compounds. AB - An innovative cryogen-free concentrator system for measurement of atmospheric trace gases at the parts per trillion level has been developed with detection by routinely used gas chromatographic methods. The first-generation system was capable of reaching a trapping temperature of -186 degrees C, while the current version can reach -195 degrees C. A Kleemenko cooler is used to create liquid nitrogen equivalent trapping conditions and eliminate the use of solid absorbents, a potential source of artifacts. The method utilizes dual-stage trapping with individual cold regions. The two stages are cooled to -20 and -175 degrees C for water management and sample enrichment, respectively. Both stages house a Silonite-coated stainless steel sample loop; the second stage loop is filled with 1-mm-diameter glass beads, which provide an inert surface area for analyte concentration. In our application, the complete system employed four channels utilizing two flame ionization detectors, one electron capture detector, and a mass spectrometer. The system was automated for unattended operation and was deployed off the New England east coast on Appledore Island to measure a suite of ambient non-methane hydrocarbons, halocarbons, alkyl nitrates, and oxygenated volatile organic compounds during the International Consortium for Atmospheric Research on Transport and Transformation field campaign in summer 2004. This robust system quantified 98 ambient volatile organic compounds with precisions ranging from 0.3 to 15%. PMID- 16255601 TI - Determination of single-cell oxygen consumption with impedance feedback for control of sample-probe separation. AB - The ability to measure chemical gradients surrounding single cells provides novel insights into several areas of cell dynamics--particularly metabolism. Detection of metabolic oxygen consumption can be achieved from a single mammalian cell using a modulated amperometric sensor in a self-referencing mode. To date, however, apart from visual cues, we do not have a reliable and cell-compatible method for determining and stabilizing the position of such probes. In this paper, we report on having successfully measured the increase in the uncompensated resistance of an electrochemical cell upon approach to single, living, biological cells, while simultaneously measuring the metabolic oxygen consumption. This was accomplished by applying an ac and a dc excitation signal to the electrode. The applied ac waveform was a 100-kHz sine wave with an amplitude of 10 mV rms, while the dc voltage applied was -600 mV. The two signals were shown not to interfere with one another. Furthermore, it is shown that the sample-probe distance can be measured for approach to single cells on the order of 10-15-microm diameter and 5-microm height, with 100-nm resolution. PMID- 16255602 TI - Molecularly imprinted polymers based on iodinated monomers for selective room temperature phosphorescence optosensing of fluoranthene in water. AB - Aiming at enhancing the advantages of traditional molecularly imprinted polymers (MIPs) for chemical sensing, a new MIP design approach introducing an internal heavy atom in their polymeric structure is described. Based on the heavy-atom effect, the novel polymer allows one to perform room-temperature phosphorescence (RTP) transduction of the analyte. The synergic combination of a tailor-made MIP recognition with a selective RTP detection is a novel concept for optosensing devices which is assessed here for simple and highly selective determination of trace amounts of fluoranthene in water. The noncovalent MIP was synthesized using the laboratory-synthesized tetraiodobisphenol A as one of the polymeric precursors and fluoranthene as template. In the presence of an oxygen scavenger, the iodide included in the polymeric structure induced efficient RTP emission from the analyte, once recognized by the MIP. The developed optosensing system has demonstrated a high specificity for fluoranthene against other polycyclic aromatic hydrocarbons. Detection limit for the target molecule was 35 ng/L (5-mL sample injections), and the linear range extended above 100 microg/L of the analyte. The polymer can be easily regenerated for subsequent sample injections (at least up to 450 cycles) with acetonitrile. The synthesized sensing material showed good stability for at least 6 months after preparation. The feasibility of monitoring fluoranthene in real samples was successfully evaluated through the analysis of five spiked river water samples. PMID- 16255603 TI - Mass spectrometry-based screening for inhibitors of beta-amyloid protein aggregation. AB - Alzheimer's disease is the most common cause of the loss of cognitive function among the elderly, and the aggregation and deposition of misfolded beta-amyloid protein (Abeta) contribute to this progressive central nervous system decline. Therefore, compounds that inhibit or even reverse Abeta aggregation might be useful for the treatment or prevention of Alzheimer's disease. To identify potential therapeutic agents for the treatment of Alzheimer's disease, a mass spectrometry-based screening assay was developed to identify and rank order compounds that inhibit the aggregation of Abeta. To carry out this assay, Abeta was incubated with a test compound at 37 degrees C for 20 h followed by ultrafiltration to separate the monomeric Abeta from its aggregates. Aliquots of the ultrafiltrate were analyzed for monomeric Abeta using positive ion electrospray mass spectrometry based on the abundance the quadruply protonated molecule of Abeta at m/z 1083. The calibration curve for Abeta was linear with a correlation coefficient (r2) of >0.99 over the range of at least 11-110 microM. The limit of detection was 0.224 ng (5.18 nM, 10-microL injection), and the limit of quantitation was 0.747 ng (17.2 nM, 10-microL injection). Based on previous reports of compounds that either bind to Abeta or are useful in treating Alzheimer's disease, melatonin, methysticin, 3-indolepropionic acid, and daunomycin were assayed and ranked in order of inhibition of Abeta aggregation. The most effective inhibitor of aggregation of Abeta protein was daunomycin followed in descending order by 3-indolepropionic acid, melatonin, and then methysticin. These data suggest that this ultrafiltration LC-MS screening assay may be used to identify potential therapeutic agents for the treatment of Alzheimer's disease based on the prevention of Abeta aggregation. PMID- 16255605 TI - Flow counterbalanced capillary electrophoresis using packed capillary columns: resolution of enantiomers and isotopomers. AB - A method with the ability to increase greatly both the resolution and efficiency of a given capillary electrophoretic system is described. This method differs from traditional capillary electrophoresis (CE) in that a counterflow is induced in the direction opposite to the electrokinetic migration of the analyte. This has the effect of extending not only the time the analytes migrate in the electric field but also the effective length and the effective applied voltage of the system. Previous work in our group with flow counterbalanced capillary electrophoresis has utilized an open tube of small inner diameter to reduce peak broadening caused by hydrodynamic flow. Narrow-diameter capillaries (5-10 microm) restricted analysis to fluorescent analytes and laser-induced fluorescence detection. The method described here uses a capillary of much larger inner diameter (75 microm) that has been packed with nonporous silica particles. The packing material reduces the amount of band broadening caused by pressure-induced flow relative to that experienced in an open tube. A larger diameter capillary allows the detection of analytes by UV absorption, not only eliminating the need to tag analytes with fluorescent tags but also allowing for the detection of a much broader range of analytes. The system was evaluated by studying the separations of several enantiomers using only beta-cyclodextrin as the chiral selector. The system was also used to resolve the two naturally occurring isotopes of bromine and to resolve phenylalanine from phenylalanine-d8. Relative to traditional CE, large improvements in resolution and separation efficiency have been achieved with this method. PMID- 16255604 TI - Quantification of cytokines involved in wound healing using surface plasmon resonance. AB - Sensing of three cytokines related to chronic wound healing, interleukin-1 (IL 1), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-alpha), with detection limits at or below 1 ng/mL in buffered saline solution and spiked cell culture medium (CCM) has been achieved. Fiber-optic surface plasmon resonance (SPR) sensors are coated with an antibody binding layer and antibodies specific to the cytokine of interest are covalently attached to this layer. To achieve such detection limits in a complex medium such as CCM, total protein content of 4 mg/mL, the use of a novel N-hydroxysuccinimide ester of 16-mercaptohexadecanoic acid (NHS-MHA) is necessary. A comparison of the detection limits for IL-6 using currently widely used CM-dextran and NHS-MHA shows an improvement by a factor of 3 using NHS-MHA. The detection limits for the monitoring of cytokines in spiked saline solutions and CCM were similar for TNF-alpha and slightly higher for IL-1 and IL-6. The detection of each cytokine in the presence of interfering agents resulted in concentration prediction well within the error of calibration. The SPR sensors are stable in CCM after 20 min of pretreatment in CCM, minimizing the reliance on a reference sensor to quantify the cytokines in complex media. This technique enables a major advancement in the field of real-time monitoring of biologically relevant molecules in complex biological fluids. PMID- 16255606 TI - Analysis of steroid conjugates in sewage influent and effluent by liquid chromatography-tandem mass spectrometry. AB - Environmental endocrine disruptors such as estrone (E1) and beta-estradiol (E2) are excreted in human urine primarily as water-soluble glucuronides and sulfates that can dissociate in wastewater treatment systems to the more active free estrogens. Measurement of the distribution and fate of the steroid conjugates and the corresponding free estrogens in treatment plants and receiving waters is critical for understanding the reproductive and developmental effects of these substances on aquatic organisms. A sensitive method to measure steroid estrogen conjugates in matrix-rich sewage influents and effluents (method detection limits ranged from 0.04 to 0.28 ng/L) has been developed using HPLC tandem mass spectrometry with electrospray ionization. The method employs extensive sample purification by selective extraction from an Oasis HLB solid-phase cartridge followed by separation by anion exchange chromatography. This purification scheme, combined with a stable isotope dilution approach, was used to overcome problems of matrix suppression of ionization and permitted selective and sensitive detection of six target conjugates of E1 and E2. Accurate quantitation was highly dependent on the method of sample preservation. Acidification of each sample (pH 2.0) was effective in preventing enzymatic or chemical decomposition of steroid conjugates in all sample types, whereas glucuronide conjugates were hydrolyzed in the presence of mercury and formalin preservatives. Measured concentrations of steroid sulfates in the influent to a sewage treatment plant were approximately 100 times greater than that of the respective steroid glucuronides, suggesting that the preponderance of glucuronides had dissociated prior to reaching the treatment plant. A small percentage of the steroid sulfates persisted through biological treatment of sewage and was measured in the effluent. Steroid conjugates that survive decomposition or bypass biological treatment of municipal wastewater are released into surface waters and may serve as a source of free steroids. PMID- 16255607 TI - Competitive binding assay for glucose based on glycodendrimer-fluorophore conjugates. AB - A new fluorescent glucose assay has been created using Alexa Fluor 647-labeled concanavalin A (Con A) and a fourth-generation PAMAM Alexa Fluor 594-labeled glycodendrimer. This assay has been shown to have a large response to glucose within the biological range and to be capable of functioning within a polymer hydrogel. In this paper, the glucose response is shown to be a single fluorophore based quenching reaction. Data showing that the sensor is fully reversible and specific through competitive binding between the dendrimer and glucose with Con A are presented. Overall, the assay is shown to have potential over the traditional dextran-based assay because it has a larger dynamic response to physiological glucose concentrations, incorporates longer wavelength dyes that improve signal penetration through dermal tissue, and provides an internal reference in the form of a nonreactive fluorescent label. PMID- 16255608 TI - Detection of single-base mutations by fluorogenic ribonuclease protection assay. AB - The ribonuclease protection assay is a generally applicable technique for the detection of known mutations. We have developed a simple and rapid method for mutation detection based on the ribonuclease protection assay using fluorescently labeled oligodeoxyribonucleotide probes. The fluorogenic ribonuclease protection (FRAP) assay uses two differently labeled oligodeoxyribonucleotides, a donor probe and an acceptor probe, to obtain a fluorescence resonance energy transfer (FRET) signal. We have utilized the FRAP assay for the detection of a single-base mutation in the YMDD motif of the hepatic B virus DNA polymerase gene. The occurrence of mismatch-selective RNA cleavage was successfully discriminated by measuring the FRET signal between the donor and acceptor probes. Moreover, mutation sensing was successfully visualized by a UV transillumination. This simple and rapid mutation sensing method should facilitate a high-throughput mutation analysis. PMID- 16255609 TI - Metal ion complexes of EDTA as solutes for density gradient ultracentrifugation: influence of metal ions. AB - In the study reported here, we study the nature of the metal ion complexes of EDTA as solute systems for analysis of lipoproteins by density gradient ultracentrifugation (DGU) by varying both the complexing metal ion and the counterion. Specifically, the sodium and cesium salts of complexes of Bi/EDTA, Pb/EDTA, Cd/EDTA, Fe/EDTA, and Cu/EDTA were chosen for this study. We show that useful gradients can be formed within a few hours beginning with a homogeneous solution. Data are presented that provide insight into the nature of how these gradients are formed from these complexes and how the selection of a specific complex can be used to enhance particular regions of the lipoprotein density profile for clinical studies. We also examine the use of equilibrium sedimentation theory to correlate the measured density profiles generated by these complexes with their molecular weight. PMID- 16255610 TI - Extraction and quantitative analysis of iodine in solid and solution matrixes. AB - 129I is a contaminant of interest in the vadose zone and groundwater at numerous federal and privately owned facilities. Several techniques have been utilized to extract iodine from solid matrixes; however, all of them rely on two fundamental approaches: liquid extraction or chemical/heat-facilitated volatilization. While these methods are typically chosen for their ease of implementation, they do not totally dissolve the solid. We defined a method that produces complete solid dissolution and conducted laboratory tests to assess its efficacy to extract iodine from solid matrixes. Testing consisted of potassium nitrate/potassium hydroxide fusion of the sample, followed by sample dissolution in a mixture of sulfuric acid and sodium bisulfite. The fusion extraction method resulted in complete sample dissolution of all solid matrixes tested. Quantitative analysis of 127I and 129I via inductively coupled plasma mass spectrometry showed better than +/-10% accuracy for certified reference standards, with the linear operating range extending more than 3 orders of magnitude (0.005-5 microg/L). Extraction and analysis of four replicates of standard reference material containing 5 microg/g 127I resulted in an average recovery of 98% with a relative deviation of 6%. This simple and cost-effective technique can be applied to solid samples of varying matrixes with little or no adaptation. PMID- 16255611 TI - Microfluidic chip for low-flow push-pull perfusion sampling in vivo with on-line analysis of amino acids. AB - Multilayer soft lithography was used to prepare a poly(dimethylsiloxane) microfluidic chip that allows for in vivo sampling of amino acid neurotransmitters by low-flow push-pull perfusion. The chip incorporates a pneumatically actuated peristaltic pump to deliver artificial cerebrospinal fluid to a push-pull perfusion probe, pull sample from the probe, perform on-line derivatization with o-phthaldialdehyde, and push derivatized amino acids into the flow-gated injector of a high-speed capillary electrophoresis-laser-induced fluorescence instrument. Peristalsis was achieved by sequential actuation of six, 200 microm wide by 15 microm high control valves that drove fluid through three fluidic channels of equal dimensions. Electropherograms with 100,000 theoretical plates were acquired at approximately 20-s intervals. Relative standard deviations of peak heights were 4% in vitro, and detection limits for the excitatory amino acids were approximately 60 nM. For in vivo measurements, push pull probes were implanted in the striatum of anesthetized rats and amino acid concentrations were monitored while sampling at 50 nL/min. o Phosphorylethanolamine, glutamate, aspartate, taurine, glutamine, serine, and glycine were all detected with stable peak heights observed for over 4 h with relative standard deviations of 10% in vivo. Basal concentrations of glutamate were 1.9 +/- 0.6 microM (n = 4) in good agreement with similar methods. Monitoring of dynamic changes of neurotransmitters resulting from 10-min applications of 70 mM K(+) through the push channel of the pump was demonstrated. The combined system allows temporal resolution for multianalyte monitoring of approximately 45 s with spatial resolution 65-fold better than conventional microdialysis probe with 4-mm length. The system demonstrates the feasibility of sampling from a complex microenvironment with transfer to a microfluidic device for on-line analysis. PMID- 16255614 TI - Molar mass and molar mass distribution of polystyrene particle size standards. AB - Monodisperse polystyrene microspheres and nanospheres are often used as particle size standards for calibration of size-measuring instruments. They are potentially useful as the mass standards for particle mass spectrometry as well. We demonstrated in this work that it is possible to achieve high-precision mass determination for single polystyrene spheres using a quadrupole ion trap. We introduced the particles into the trap by laser-induced acoustic desorption and probed them with light scattering. Mass-to-charge ratios of the individual particles were determined from applied trap-driving frequencies, voltage amplitudes and the observed starlike oscillatory trajectories projected on the radial plane. Creation of one-electron differentials through charge-state changes by electron bombardment allowed determination for the absolute mass of a single trapped particle to a precision better than 0.1%. Both molar mass and molar mass distribution were deduced from a large number of measurements for NIST polystyrene particle size standards (SRMs 1690 and 1691). Our results are in excellent agreement with the size measurement for the 0.895-microm spheres (NIST SRM 1690), but a small discrepancy (4%) in number-average mass was found for the 0.269-microm spheres (NIST SRM 1691). PMID- 16255612 TI - Spraying enzymes in microemulsions of AOT in nonpolar organic solvents for fabrication of enzyme electrodes. AB - A new technique suitable for automated, large-scale fabrication of enzyme electrodes by air-spraying enzymes in organic inks is presented. Model oxidoreductases, tyrosinase (Tyr) and glucose oxidase (GOx), were adapted to octane-based ink by entrapment in a system of reverse micelles (RM) of surfactant AOT in octane to separate and stabilize the catalytically active forms of the enzymes in nonpolar organic media. Nonpolar caoutchouk polymer was also used to create a kind of "dry micelles" at the electrode/solution interface. Enzyme/RM/polymer-containing organic inks were air-brushed onto conductive supports and were subsequently covered by sprayed Nafion membranes. The air brushed enzyme electrodes exhibited relevant bioelectrocatalytic activity toward catechol and glucose, with a linear detection range of 0.1-100 microM catechol and 0.5-7 mM glucose; the sensitivities were 2.41 A M(-1) cm(-2) and 2.98 mA M( 1) cm(-2) for Tyr and GOx electrodes, respectively. The proposed technique of air brushing enzymes in organic inks enables automated construction of disposable enzyme electrodes of various designs on a mass-production scale. PMID- 16255613 TI - Creation of a P450 array toward high-throughput analysis. AB - The rapid metabolism testing of many new chemical entities enables unsuitable candidates to be eliminated from consideration at an early stage of the drug discovery process. We have developed a P450 array toward high-throughput analysis of P450-mediated metabolic reaction. The microsomes containing expressed human P450 enzymes were immobilized on the microassay plate using sol-gel chemistry. A thin-film hydrogel containing microsomes was fabricated using aqueous silicate as a starting material. The TEM image clearly showed that the nanoclusters derived from the silicate formed branched chains, and microsomes were entrapped in the silica network. The different P450 isozymes were immobilized on the microassay plate, and the metabolites by each isozyme were visualized as fluorescent images, which creates opportunity for the inhibitor assays. This method offers several advantages over use of conventional enzyme preparations, including increased storage stability, ease of product isolation from the incubation mixture, and the ability to recover and reuse the enzyme. Because this methodology enabled the development of assay system using P450 that is unstable and involves other enzymes for its function, it can be applicable to various screening assays that require complicated reactions involving many biological components. PMID- 16255615 TI - Design for DNA separation medium using bacterial cellulose fibrils. AB - In this paper, we present a novel DNA separation medium using bacterial cellulose fibrils. Bacterial cellulose has an intrinsic three-dimensional micrometer- to nanometer-scale network structure. Addition of this material to a low concentration polymer solution (<5 cP) enables high-resolution electrophoretic separation of DNA, even for fragments of 10-100-bp or single-nucleotide polymorphism. The newly designed medium consists of a double mesh: a 10-nm flexible mesh derived from a conventional polymer medium containing 10-nm to 1 microm rigid pores made up of 10-microm bacterial cellulose fragments. PMID- 16255616 TI - Chromatography on self-assembled carbon nanotubes. AB - Stationary phases that provide high resolutions and are stable at high temperatures are of significant importance in chromatographic analysis. Carbon nanotubes (CNTs) are known to have high thermal and mechanical stability and have the potential to be high-performance separation media that utilize the nanoscale interactions. Here, we report the first application of self-assembled CNTs in long capillary tubes for the development of gas chromatography columns. A film of CNTs was deposited by chemical vapor deposition (CVD) to form the stationary phase in the open tubular format. High-resolution separation of a number of compounds has been achieved. Altering the CVD conditions can vary the thickness and the morphology of the CNT film, which opens the possibility of selectivity tuning. The ability to fabricate long tubes coated with CNTs can be readily employed in other gas- and liquid-phase separations as well. PMID- 16255617 TI - Mass spectrometric determination of 234U/238U ratio with improved precision. AB - A new mass spectrometric method is proposed for measurement of 234U/238U ratio with a single Daly electron multiplier detector using the general peak jump method. The method is based on precise measurement of the 235U/238U ratio and 234U/235U ratio, which are used to calculate the 234U/238U ratio using the equation 234U/238U = 235U/238U x 234U/235U. The results show a significant improvement, i.e., more than 35 times better precision in measuring the (234)U/(238)U ratio with this method (sigma = 2.9 x 10(-8)) as compared to direct measurement of 234U/238U (sigma = 1.1 x 10(-6)). The method widens the applicability of the single collector system, and it will potentially be helpful to improve the precision in the case of the static multicollector system also. PMID- 16255619 TI - A systematic analytic approach to pandemic influenza preparedness planning. PMID- 16255620 TI - Barriers facing junior doctors in rural practice. AB - INTRODUCTION: Early postgraduate, or junior doctors, are still required to practise in rural and remote communities, and they continue to face numerous issues and difficulties. Within the hospital setting, exposure to rural practice appears to be very limited during internship, and also to some extent, during the second postgraduate year and beyond. This is a major issue for those required to undertake country relieving, rural terms or who will be bonded to rural and remote practice for several years after internship. This research investigated the current issues and difficulties faced by junior doctors, required to undertake rural and remote practice in Queensland, Australia. METHODS: An exploratory study was undertaken. Primary data were collected through semi structured interviews held with key stakeholders. Stakeholders included: directors of clinical training; medical educators; junior doctors; rural practitioners; academic rural practitioners; and medical administrators. Of the 23 people approached, a total of 19 agreed to be interviewed. The response rate was 82.6%. RESULTS: Similar to the issues identified in the literature, there are currently a number of barriers influencing the ability of junior doctors to practise competently and confidently when undertaking practice in rural and remote communities. Minimal clinical experience, lack of supervision and on-site support, inadequate orientation and uninformed expectations, limited access to relevant education, and the influence of isolation, results in an overall lack of preparation both professionally and personally. When asked, respondents supported the identification of core skills and knowledge, and integration of these and other issues affecting rural practice, into their hospital-based programs. Current hospital-based education and training programs were not adequately preparing junior doctors for rural and remote practice. It was commented that orientation and education, with a rural emphasis, could assist junior doctors in their preparation for country relieving, rural terms and longer placements. CONCLUSIONS: Data collected in this study have confirmed that junior doctors are still being sent to undertake country relieving in their second postgraduate year. Hence, the issues remain for junior doctors when undertaking practice in rural or remote communities, including country relievers and scholarship holders. Results from this study suggest that prior recommendations have not been fully implemented. A recommendation is that initiatives at the undergraduate level, including increasing rural exposure and integration of rural context into training, be further developed at the early postgraduate level. Core rural competencies should be identified and realistic preparation and support strategies put in place in the hospital setting. This will further the effort to prepare junior doctors for rural practice and minimise some of the barriers currently experienced. PMID- 16255621 TI - Australian rural midwives: perspectives on Continuing Professional Development. AB - INTRODUCTION: Continuing Professional Development (CPD) provides an important counter to workforce pressures affecting rural midwives; however, there is a lack of information about how rural midwives understand and perceive CPD and how this is situated in the practising and social context. This research aimed to explore rural midwives' experiences and perceptions of CPD in context. METHODS: A qualitative approach gathered focus group data on the beliefs, opinions and perceptions of a total of 52 rural midwives (nine focus groups), across three Australian states: Western Australia, Victoria and Tasmania. The focus groups were taped and transcribed verbatim and data was analysed thematically using an inductive approach, with the aid of an NVivo (QSR Software, Durham, UK) computer program. RESULTS: Four key themes emerged from the data: midwives' views of CPD; their motivations for undertaking CPD; the choices they make around CPD; and how context factors facilitate their involvement in CPD. Congruence with issues evident in the literature were: the difficulties associated with role diversity, the need for acquiring key skills before engaging in CPD, and the importance of a culture supportive of ongoing learning. CONCLUSIONS: CPD can be considered an important strategy for the retention and professionalism of midwives. The study findings helped fill a gap in the literature about rural and regional Australian midwives' perspectives on CPD. PMID- 16255622 TI - Genomic microarrays in cancer molecular diagnostics: just biomarker discovery tools or future bedside clinical assays? PMID- 16255623 TI - BioNanoMedicine: a nanotechnology platform for the 21st century. PMID- 16255625 TI - Chips to Hits: microarray and microfluidic technologies for high-throughput analysis and drug discovery. September 12-15, 2005, MA, USA. PMID- 16255627 TI - BioArrays Europe 2005. September 27-30, 2005, London, UK. PMID- 16255626 TI - IBC's 4th Annual Conference on Molecular Diagnostics. September 13-14, 2005, Convention and Expo Center, Boston, MA, USA. PMID- 16255628 TI - Liquid-based cytology for cervical cancer screening. AB - In multiple studies during the last decade, liquid-based cytology for cervical cancer screening has been shown to increase the detection rate for preneoplastic squamous intraepithelial lesions equal to or greater than the conventional Papanicolaou (Pap) smear method. Liquid-based collection and processing provide more representative cervical sampling than conventional smearing of the specimen on a glass slide. Currently, there are two test methodologies that are widely marketed and available to clinical laboratories, health systems and clinicians that undertake cervical cytology. The purpose of this article is to provide an overview of the methodology and performance of SurePath Liquid-Based Pap Test in cervical cytology screening. The SurePath liquid-based Pap test significantly reduces the unsatisfactory rate of Pap test slides, and detects a significantly higher number of low- and high-grade squamous lesions when compared with the conventional Pap smear technique. Biopsy confirmation shows that this increased detection does not come at a cost of decreasing specificity, and sensitivity for histologic dysplasia is equal to or greater than the best available data for the conventional Pap method. The SurePath collection vial provides residual cellular material for adjunctive out-of-the-vial molecular testing, including sexually transmitted diseases and oncologic biomarkers associated with cervical carcinoma. Finally, SurePath slides can be placed on an automated cervical cytology screening device (FocalPoint), thus providing improved disease detection and enhanced laboratory productivity. PMID- 16255629 TI - Analysis of nipple aspirate fluid for diagnosis of breast cancer: an alternative to invasive biopsy. AB - Over 40,000 women in the USA will die this year of breast cancer. Current generally accepted techniques to detect breast cancer are limited to breast examination and mammography. Abnormalities found by these techniques require an invasive needle or surgical biopsy to determine if cancer is present. The author's ultimate goal is to determine if a woman has breast cancer without the need for invasive biopsies, and do this before the abnormality is detectable by standard screening techniques. Herein, the technology is reviewed as it was, as it is today, and its future potential is discussed. PMID- 16255630 TI - Evolution of prenatal genetics: from point mutation testing to chromosomal microarray analysis. AB - Molecular genetic testing involves DNA analysis using various methods for the purpose of diagnosing genetic disorders. In the prenatal DNA diagnostic setting, fetal DNA is usually tested for a specific single-gene disorder for which the fetal risk is 25% or more. In contrast, cytogenetic testing is often used to detect fetal chromosomal abnormalities in cases that involve a wider range of indications. Classic cytogenetic and DNA-based testing methods provide a range of aberrations detected with different levels of genomic resolution. More recently developed molecular cytogenetic methods provide powerful tools to bridge the technical divide between these related areas. One such hybrid method is microarray-based comparative genomic hybridization. Chromosomal microarray analysis has been applied to clinical testing for unbalanced gains or losses of genomic regions associated with genetic disorders. This technology is poised to have a substantial impact on clinical genetics, including prenatal genetic testing. PMID- 16255631 TI - Calcium phosphate nanoparticles: second-generation nonviral vectors in gene therapy. AB - Adverse effects of viral vectors, instability of naked DNA, cytotoxicity and low transfection of cationic lipids, cationic polymers and other synthetic vectors are currently severe limitations in gene therapy. In addition to targeting a specific cell type, an ideal nonviral vector must manifest an efficient endosomal escape, render sufficient protection of DNA in the cytosol and help provide an easy passage of cytosolic DNA to the nucleus. Virus-like size calcium phosphate nanoparticles have been found to overcome many of these limitations in delivering genes to the nucleus of specific cells. This review has focused on some applications of DNA-loaded calcium phosphate nanoparticles as nonviral vectors in gene delivery, and their potential use in gene therapy, as well as highlighting the mechanistic studies to probe the reason for high transfection efficiency of the vector. It has been demonstrated that calcium ions play an important role in endosomal escape, cytosolic stability and enhanced nuclear uptake of DNA through nuclear pore complexes. The special role of exogenous calcium ions to overcome obstacles in practical realization of this field suggests that calcium phosphate nanoparticles are not 'me too' synthetic vectors and can be designated as second generation nonviral vectors for gene therapy. PMID- 16255632 TI - Telomerase: is it the future diagnostic and prognostic tool in human cancer? AB - A number of methods exist to detect levels of telomerase activity and the presence of telomerase subunits in a variety of tissues. As telomerase activation seems to be an important step in tumorigenesis, accurate detection of the presence and activity of the enzyme and its subunits is vital. The original method of detecting telomerase activity was developed by Kim and coworkers in 1994, and was termed the telomeric repeat amplification protocol. This assay led to a staggering increase in the number of telomerase-associated publications in scientific journals (85 publications from 1974-1994, 5063 publications from 1994 2004). A number of methods have been described to detect telomeres and to measure their length, with the standard measurement of telomere length performed using a modification of the Southern blot protocol. RNA in situ hybridization can be performed to detect levels of the RNA component of telomerase, and standard in situ hybridization and immunohistochemistry can be applied to examine expression levels and localization of the catalytic subunit of the enzyme. Reverse transcriptase PCR has also been applied to assess expression levels of the telomerase components in various tissues. This review provides a synopsis of telomeres, telomerase, telomerase and cancer, and finally, methods for the detection of telomerase in cancer. PMID- 16255633 TI - Digital karyotyping technology: exploring the cancer genome. AB - Identifying gene-specific alterations in cancer genomes has revealed molecules that are causal effectors of carcinogenesis and specific targets for cancer molecular diagnosis and molecular-based cancer therapies. Whole-genome analyses of many cancer genomes at the resolution of single genes is thus a desirable yet incompletely realized goal that could expedite progress in cancer diagnosis and treatment. Although methods for routine whole-genome sequencing or high resolution epigenetic measurements are currently under development, high resolution measurements of gene copy number, or 'gene dosage', are now underway in several laboratories. Digital karyotyping, array comparative genomic hybridization, and single nucleotide polymorphism arrays are techniques that have the potential to detect gene amplification, homozygous deletion and loss of heterozygosity at or below the average length of single genes. Recently, digital karyotyping of a small number (<20) of colon and brain cancer genomes has revealed tumor cases with significant genetic dosage alterations affecting few and, in some cases, only one complete gene. These experiments suggest that gene specific gene dosage alterations may be sufficiently frequent to enable the identification of promising tumor gene candidates in small-scale experiments. The purpose of this review is to describe our understanding of cancer as a genetic disease, review the basic principles, methodologies and interpretational issues of traditional and high-resolution whole-genome screens, and describe the potential of our first detailed look at whole cancer genomes for progress in the understanding and treatment of cancer. PMID- 16255635 TI - Pyrosequencing: nucleotide sequencing technology with bacterial genotyping applications. AB - Pyrosequencing is a relatively new method for real-time nucleotide sequencing. It has rapidly found applications in DNA sequencing, genotyping, single nucleotide polymorphism analysis, allele quantification and whole-genome sequencing within the areas of microbiology, clinical genetics and pharmacogenetics. It is fast becoming a real alternative to the traditional Sanger sequencing method although, at present, read lengths are normally limited to approximately 70 nucleotides. The pyrosequencing method involves four main stages: first, target DNA is amplified using PCR; second, double-stranded DNA is converted to single-stranded DNA templates; third, oligonucleotide primers are hybridized to a complementary sequence of interest; and, finally, the pyrosequencing reaction itself, in which a reaction mixture of enzymes and substrates catalyses the synthesis of complementary nucleotides. Data are shown as a collection of signal peaks in a pyrogram. Pyrosequencing is increasingly used for bacterial detection, identification and typing, and, recently, a commercial system became available for the identification of bacterial isolates. Pyrosequencing can also be partially or fully automated, thus enabling the high-throughput analysis of samples. Wider use of pyrosequencing may occur in the future if longer nucleotide reads are made possible, which will enable its expansion into larger nucleotide sequencing such as multilocus sequence typing and whole-genome sequencing. PMID- 16255634 TI - Molecular classification and biomarker discovery in papillary thyroid carcinoma. AB - Papillary thyroid carcinoma (PTC) is the most common thyroid malignancy, with an incidence of approximately 22,000 cases in 2004 in the USA. Incidence is increasing, with a global estimate of half a million new cases this year. PTC is found in a variety of morphologic variants, usually grows slowly and is clinically indolent, although rare, aggressive forms with local invasion or distant metastases can occur. In recent years, thyroid cancer has been at the forefront of molecular pathology as a result of the consequences of the Chernobyl disaster and the recognition of the role of Ret/PTC rearrangements in PTC. Nonetheless, the molecular pathogenesis of this disease remains poorly characterized. In the clinical setting, benign thyroid nodules are far more frequent, and distinguishing between them and malignant nodules is a common diagnostic problem. It is estimated that 5-10% of people will develop a clinically significant thyroid nodule during their lifetime. Although the introduction of fine-needle aspiration has made PTC identification more reliable, clinicians often have to make decisions regarding patient care on the basis of equivocal information. Thus, the existing diagnostic tools available to distinguish benign from malignant neoplasms are not always reliable. This article will critically evaluate recently described putative biomarkers and their potential future role for diagnostic purposes in fine-needle aspiration cytology samples. It will highlight the evolution of our understanding of the molecular biology of PTC, from a narrow focus on specific molecular lesions such as Ret/PTC rearrangements to a pan-genomic approach. PMID- 16255636 TI - HIV/AIDS among Asians and Pacific Islanders in the United States. PMID- 16255637 TI - Epidemiology of HIV/AIDS among Asians and Pacific Islanders in the United States. AB - Although the percentage of overall AIDS diagnoses remains low among Asian and Pacific Islanders (APIs) in the United States compared with other racial/ethnic groups, research on API risk behaviors and health status suggest that the low number of AIDS cases may not provide a full picture of the epidemic and issues faced by this understudied and underserved population. Data from national HIV/AIDS surveillance systems and the Behavioral Risk Factor Surveillance System (BRFSS) were examined to delineate the magnitude and course of the HIV/AIDS epidemic among APIs in the United States. Same-sex sexual activity is the main HIV risk for API men, whereas heterosexual contact is for API women. APIs are significantly less likely to report being tested for HIV despite the fact that a similar proportion of APIs and other racial/ethnic groups reported having HIV risk in the past 12 months. Given the enormous diversity among APIs in the United States it is important to collect detailed demographic information to improve race/ethnicity and HIV risk classification, conduct better behavioral and disease monitoring for informing prevention planning, and addressing cultural, linguistic, economic and legal barriers to HIV prevention among APIs. PMID- 16255638 TI - Substance use, substance choice, and unprotected anal intercourse among young Asian American and Pacific Islander men who have sex with men. AB - Substance use has been shown to be an important factor associated with having unprotected anal intercourse (UAI) among Asian and Pacific Islander (API) men who have sex with men (MSM). However, little is known about which substances are used in conjunction with sexual activity and whether having UAI varies by substance choice in this population. From January 2000 to September 2001, we sampled API MSM aged 18-29 years from 30 gay-identified venues in San Francisco, California, and interviewed 496 API men face-to-face using a standardized questionnaire. Overall, 47% of the sample reported UAI in the past 6 months. During the same time period, 32% and 34% reported being "high" or "buzzed" on alcohol and drugs during sex, respectively. The most common drugs used in conjunction with sex were methylenedioxymethamphetamine ("ecstasy"; 19%), followed by marijuana (14%), inhalant nitrites ("poppers"; 11%), and crystal methamphetamine ("crystal"; 10%). In a multivariate model, we observed associations between UAI and being high or buzzed on ecstasy (odds ratio [OR] = 2.62; 95% confidence interval [CI] = 1.37, 5.02) and poppers during sex (OR = 3.29; 95% CI = 1.50, 7.25). However, being high or buzzed on alcohol, marijuana, gamma-hydroxybutyrate (GHB), and crystal methamphetamine during sex had no association with UAI. One third of sampled young API MSM used drugs or alcohol during sex. The co-occurrence of ecstasy and popper use and unprotected sex underscores the need to develop HIV prevention programs focusing on particular drugs. PMID- 16255639 TI - Sexual risk behavior and substance use among a sample of Asian Pacific Islander transgendered women. AB - We examined the prevalence and correlates of HIV-related sexual risk and substance use behaviors among Asian Pacific Islander (API) male-to-female (MTF) transgendered individuals, referred to here as API transgendered women. As part of a larger study on HIV risk among transgendered women of color (Nemoto, Operario, Keatley, Han, & Soma, 2004), a sample of 110 API transgendered women in San Francisco completed individual interviews, of which 13% reported being HIV positive. In the past 30 days, one fifth of the sample engaged in unprotected receptive anal intercourse (URAI) with any male partner, nearly one half had sex while under the influence of substances, and over half used illicit drugs. In multivariate models, URAI was associated with commercial sex work (odds ratio [OR] = 4.23, 95% confidence interval [CI] = 1.10, 16.25) and previous attempted suicide (OR = 5.83, 95% CI = 1.02, 33.44). Sex under the influence of substances was associated with commercial sex work (OR = 3.35, 95% CI = 1.11, 10.13) and having a college degree (OR = 5.32, 95% CI = 1.34, 21.18). Illicit drug use was associated with commercial sex work (OR = 7.15, 95% = 2.26, 22.63). Findings suggest that API MTF transgenders are on the front line of HIV risk for the API community, and provide insight into factors within this group that might contribute to unsafe sex and substance use. PMID- 16255640 TI - Risk behaviors among Asian women who work at massage parlors in San Francisco: perspectives from masseuses and owners/managers. AB - This study investigates cognitive, cultural, and contextual factors that influence HIV-related risk behaviors among Asian women who engage in sex work at massage parlors in San Francisco. Focus groups and qualitative interviews were conducted for Vietnamese and Thai masseuses and massage parlor owners/managers. Economic pressure as well as subjective evaluation of customers for the risk of HIV/STD infection increase unprotected sexual behaviors among Asian masseuses. Massage parlor owners/managers do not establish a clear policy for condom use at their parlors. Consequently, male customers often manipulate their intention not to use a condom while negotiating with masseuses. HIV/STD prevention intervention programs should address specific risks and needs for Asian masseuses, owners/managers, and male customers in order to promote health and well being among Asian masseuses who have been neglected by public health and social services. PMID- 16255641 TI - Sex, HIV risks, and substance use among Asian American college students. AB - In this study of 248 predominantly heterosexual Asian American college students, we found some HIV risks: lifetime prevalence of unprotected sex (37%), alcohol before sex (23.8%), and drug use before sex (6.0%). The prevalence of lifetime anal sex is only 9%, but 90.48% of those who have ever had anal sex did so without a condom. The Sexual Risk Indices, measured with a 13 item risky sexual behavior checklist, are positively associated with age. Most students have inadequate HIV knowledge. Acculturation is positively associated with the 30-day HIV Sexual Risk Index and HIV Knowledge Score. Preference for speaking English at home and for American entertainment is associated with higher likelihood of sexual activities, safe or unsafe. Lifetime Sexual Risk Index is also associated with 30-day and lifetime alcohol use. Lifetime drug use is associated with 30-day and lifetime Sexual Risk Indices. Alcohol before use in the lifetime also reliably predicts unprotected sex in the lifetime. PMID- 16255642 TI - The health belief model, sexual behaviors, and HIV risk among Taiwanese immigrants. AB - In this first investigation of Taiwanese sexual behaviors in the United States, 144 Taiwanese students completed an online anonymous survey. Demographics, health belief model (HBM) constructs, and acculturation were examined as predictors of sexual behaviors over the last year. Analyses indicated that participants who reported a higher number of sexual partners and more frequent sexual intercourse tended to be more educated and more likely to be nonheterosexual. The HBM constructs, as a set, reliably predicted participants' sexual behaviors. Self efficacy was the strongest predictor within the HBM. Furthermore, acculturation moderated the predictive power of the HBM with respect to intercourse frequency. The main limitation of the study is that the measure of HBM, which was not designed to target Asian immigrants, was psychometrically poor. The results suggest self-efficacy is a target for behavioral change, acculturation may need to be incorporated into the HBM, and more culturally sensitive measures need to be developed. PMID- 16255644 TI - Synthetic biology. AB - Chemistry is a broadly powerful discipline in contemporary science because it has the ability to create new forms of the matter that it studies. By doing so, chemistry can test models that connect molecular structure to behaviour without having to rely on what nature has provided. This creation, known as 'synthesis', began to be applied to living systems in the 1980s as recombinant DNA technologies allowed biologists to deliberately change the molecular structure of the microbes that they studied, and automated chemical synthesis of DNA became widely available to support these activities. The impact of the information that has emerged has made biologists aware of a truism that has long been known in chemistry: synthesis drives discovery and understanding in ways that analysis cannot. Synthetic biology is now setting an ambitious goal: to recreate in artificial systems the emergent properties found in natural biology. By doing so, it is advancing our understanding of the molecular basis of genetics in ways that analysis alone cannot. More practically, it has yielded artificial genetic systems that improve the healthcare of some 400,000 Americans annually. Synthetic biology is now set to take the next step, to create artificial Darwinian systems by direct construction. Supported by the National Science Foundation as part of its Chemical Bonding program, this work cannot help but generate clarity in our understanding of how biological systems work. PMID- 16255643 TI - Chinese and South Asian religious institutions and HIV prevention in New York City. AB - Religious institutions in Asian immigrant communities are in a unique position to confront the challenges of the HIV epidemic for the populations they serve. However, there has been little research on whether these institutions are willing or able to take a role in HIV prevention. This article reports on findings from a qualitative study of three Asian immigrant religious institutions in New York City (a Buddhist temple, a Hindu temple, an Islamic center/mosque) that are part of a larger study of Asian immigrant community institutions and their response to the HIV epidemic. Several prominent themes arose that formed the basis of a preliminary theoretical framework describing the way Asian immigrant religious institutions may evaluate their role in HIV prevention. The interview data indicate that the institutions take a stance of "conservative innovation," weighing their role as keepers of morality and religious tradition against the changing needs of their communities and then adjusting their practices or positions incrementally (to varying degrees) to stay responsive and relevant. PMID- 16255646 TI - IL-7 as a potential therapy for HIV-1-infected individuals. AB - Highly active antiretroviral therapy (HAART), although effective in ameliorating the quality of life of HIV-1-infected individuals and their survival, has not been able to eradicate HIV-1. In fact, when HAART is interrupted, HIV-1 plasma viral load rebounds from viral reservoirs such as resting CD4+ T lymphocytes, monocytes and macrophages, remaining a major obstacle in attempting HIV eradication. Different therapeutic strategies have been attempted, such as structured treatment interruption (STI), immunotherapy (interleukin [IL]-2 and anti-CD3 antibodies [e.g., OKT3]), to try to stimulate HIV-1 out of latency along with antiretroviral intensification therapy. IL-7, a pleiotropic cytokine, bears diverse immune properties and plays a major role in T cell homeostasis. Moreover, IL-7 has recently been investigated as a possible immune adjuvant as well as a viral strain-specific inducer of HIV-1 replication. In fact, IL-7 was shown not only to be more effective than IL-2 in stimulating HIV-1 replication from resting CD4+ T lymphocytes ex vivo, but also to selectively induce a specific HIV-1 viral strain as compared with IL-2, suggesting the potential need for different viral inducers if complete eradication is to be achieved. In this present review, different immunological and virological properties of IL-7 are discussed, along with the possibility of its use as part of a combined antiretroviral-immune rationally based HIV-1 eradication approach. PMID- 16255645 TI - Current status of animal-to-human transplantation. AB - The transplantation of animal organs into humans as a way of treating organ failure has been pursued for 100 years. Clinical xenotransplantation, as such, has always failed because the transplanted organ is rejected by the recipient. Recent advances in transplant immunology have revealed some mechanisms underlying the rejection of xenografts, and these discoveries have sparked efforts to use genetic engineering of animals and therapeutics directed at the recipient to overcome this problem. This paper reviews the current understanding of the mechanisms of xenograft rejection and efforts to overcome rejection and other hurdles. PMID- 16255647 TI - Individualised cancer therapeutics: dream or reality? Therapeutics construction. AB - The analysis of DNA microarray and proteomic data, and the subsequent integration into functional expression sets, provides a circuit map of the hierarchical cellular networks responsible for sustaining the viability and environmental competitiveness of cancer cells, that is, their robust systematics. These technologies can be used to 'snapshot' the unique patterns of molecular derangements and modified interactions in cancer, and allow for strategic selection of therapeutics that best match the individual profile of the tumour. This review highlights technology that can be used to selectively disrupt critical molecular targets and describes possible vehicles to deliver the synthesised molecular therapeutics to the relevant cellular compartments of the malignant cells. RNA interference (RNAi) involves a group of evolutionarily conserved gene silencing mechanisms in which small sequences of double-stranded RNA or intrinsic antisense RNA trigger mRNA cleavage or translational repression, respectively. Although RNAi molecules can be synthesised to 'silence' virtually any gene, even if upregulated, a mechanism for selective delivery of RNAi effectors to sites of malignant disease remains challenging. The authors will discuss gene-modified conditionally replicating viruses as candidate vehicles for the delivery of RNAi. PMID- 16255648 TI - Delivery of neurotrophic factors and therapeutic proteins for retinal diseases. AB - Neurotrophic factors have the ability to protect and initiate growth of neurons. In the central nervous system, neurotrophic factors are neuroprotective in a wide range of disease states. Similarly, disease pathologies of the neurosensory retina respond favourably in animal models of retinitis pigmentosa, macular degeneration, glaucoma and others. With advances in drug delivery and cell therapy, an almost universal increase in efficacy is being realised. Now, repeated injections of neurotrophic factors are being replaced by controlled delivery of cell-mediated factor secretion, reducing the number of potential acute side effects. Tissue engineering strategies in conjunction with gene modulated protein therapy or gene transfer are creating a unique treatment niche and are quickly gaining acclaim in the clinic. This review surveys the founding and current work on neurotrophic factors such as CNTF, BDNF, GDNF, LEDGF, PEDF and others. Ongoing clinical trials and successful preclinical studies are summarised as well. PMID- 16255649 TI - Anticalins as an alternative to antibody technology. AB - Anticalins are a class of engineered ligand-binding proteins that are based on the lipocalin scaffold. The lipocalin protein architecture is characterised by a compact, rigid beta-barrel that supports four structurally hypervariable loops. These loops form a pocket for the specific complexation of differing target molecules. Natural lipocalins occur in human plasma and body fluids, where they usually function in the transport of vitamins, steroids or metabolic compounds. Using targeted mutagenesis of the loop region and biochemical selection techniques, variants with novel ligand specificities, both for low-molecular weight substances and for macromolecular protein targets, can be generated. Due to their small size, typically between 160 and 180 residues, robust tertiary structure and composition of a single polypeptide chain, such 'anticalins' provide several advantages over antibodies concerning economy of production, stability during storage, faster pharmacokinetics and better tissue penetration. At present, anticalins offer three major mechanisms for therapeutic application: (i) as antidotes, by quickly removing toxic or otherwise irritating compounds from the human body; (ii) as antagonists, for example, by binding to cellular receptors and blocking them from interaction with their natural signalling molecules; (iii) as tissue-targeting vehicles, by addressing toxic molecules or enzymes to disease-related cell surface proteins. PMID- 16255650 TI - RNAi as an antiviral therapy. AB - There are a dozen or so viruses that will continue to be a serious global health threat for many years to come, mainly due to their chronic nature. These include hepatitis C virus (HCV), human papillomavirus viruses (HPVs), West Nile virus and human herpes viruses (i.e., HSV, CMV, EBV, HHV-8, etc.). However, HIV-1 infections will remain at the top of the list due to its high prevalence and the significant mortality and morbidity from AIDS. The development of a suitable vaccine against HIV-1 remains an important area of public interest. The initial hope of identifying the specific anti-HIV-1 antigenic epitopes that can protect HIV-1-infected individuals and serve as a potential vaccine has been replaced by the realisation that we have yet to identify a clear correlation of protective immunity against HIV-1 infection. Understanding the anti-HIV-1 protective factors and their potential role in the development of a vaccine or inexpensive therapy remains one of the major obstacles in HIV-1 research. In the last quarter century -since the realisation of AIDS--previous studies have established that the role of humoral or cellular immune responses in protecting human hosts against HIV-1 have been inconclusive. Moreover, most of the publicized and awaited clinical trials on vaccines have failed. The recent discovery of RNA interference (RNAi) has raised the possibility of developing a new generation of vaccines that can stymie human viruses, particularly HIV-1 replication at various stages of its life cycle at the intracellular level. Various transcripts in the HIV-1 life cycle can be targeted, and specific small double-stranded RNAs (small interfering RNAs) can be developed against these HIV-1-specific targets. However, some recent data suggests that RNAi-based therapeutics against this virus should be viewed with strong caution. Specifically, there are multiple factors that make HIV-1 a difficult infection to 'cure' because of HIV-1 latency. The changing nature of HIV-1 genomes and the possible presence of microRNAs within the HIV-1 genes can suppress RNAi directed against HIV-1 gene targets. Thus, HIV-1 would be a difficult epidemic to overcome by RNAi-based therapeutics. PMID- 16255651 TI - Meningococcal conjugate vaccines. AB - Disease caused by Neisseria meningitidis is associated with high mortality rates and significant sequelae. Polysaccharide meningococcal vaccines have been available for > 20 years, and have been used in travellers to control outbreaks, and in some countries for adolescents entering college, although they provide only a short duration of immunity and do not produce a herd effect. Monovalent and quadrivalent (A, C, Y, W-135) meningococcal conjugate vaccines have recently been licensed and endorsed for use in infants, children, adolescents and adults in various countries. Among the key features of the new meningococcal conjugate vaccines are stimulation of both B cell-dependent and T cell-dependent immune responses, induction of immunological memory and booster effects, longer-term protection, reduction of nasopharyngeal carriage of N. meningitidis, and herd immunity. Large, randomised, double-blind studies in adults, adolescents, children and infants have demonstrated the immunogenicity and safety of the new meningococcal conjugate vaccine formulations. PMID- 16255652 TI - Adalimumab: an anti-TNF agent for the treatment of psoriatic arthritis. AB - Tumour necrosis factor (TNF) has a central role in the pathogenesis of psoriatic arthritis (PsA). Adalimumab (Humira, Abbott Laboratories) is the first fully human, recombinant IgG1 monoclonal antibody that specifically targets human TNF. Adalimumab blocks the interaction of TNF with the p55 and p75 cell surface TNF receptors, thereby neutralising the activity of this cytokine. In clinical trials of patients with PsA, adalimumab significantly reduced both joint and skin symptoms. In addition, structural changes were inhibited, and statistically and clinically significant improvements in measures of disability and quality of life were observed. Adalimumab was generally safe and well tolerated. PMID- 16255653 TI - Treatment of psoriatic arthritis with etanercept, a tumour necrosis factor antagonist. AB - Psoriatic arthritis (PsA) is a chronic spondylarthritis that occurs in approximately 23% of plaque psoriasis sufferers. Traditional treatments for rheumatoid arthritis have been used as the first therapeutic approach to treat this inflammatory disease, which has both joint and skin manifestations. However, due to the inefficiency of current disease-modifying antirheumatic drugs and non steroidal anti-inflammatory drugs in stopping the progression of the joint disease, biologics have emerged as a hopeful alternative to PsA therapy. Etanercept was the first approved tumour necrosis factor-alpha (TNF-alpha) inhibitor for reducing the signs and symptoms of PsA, as well as preventing the progression of the disease. Etanercept is a fully human, soluble, dimeric fusion protein that has the ability to bind to two molecules of TNF, thereby rendering them biologically inactive. Two clinical trials have demonstrated that etanercept is generally a safe, efficacious and well-tolerated biologic therapy for the treatment of PsA. PMID- 16255655 TI - Drug interactions and fluoxetine: a commentary from a clinician's perspective. AB - This commentary suggests that clinicians require an additional perspective to that expressed in Wernicke's review. Clinicians must consider the relative risks, because even small differences may be a reason to adjust usage of a particular drug. As early as 1993, reviews noted a 'serious' interaction potential. Also, the product information requires supplementation because it does not provide the emphasis on predictable interactions that clinicians require. An extensive basis in experimental pharmacology provides the foundations for our knowledge of cytochrome P450 interactions which predict the effects on the levels of other drugs. This work has been confirmed in human studies. Adverse outcomes related to fluoxetine interactions are relatively frequent and sometimes fatal. Patients may suffer serious and irreversible ill-effects from the increased risk of many manifestations of toxicity, and the cost of resultant medico-legal settlements is considerable. A clinician who balances risks may use fluoxetine less frequently. PMID- 16255656 TI - Interactions between herbal remedies and antirheumatic drugs. AB - Many patients with rheumatological conditions use herbal remedies as an adjunct to their conventional antirheumatic medication, often without seeking advice. Herbal remedies are exempt from the usual drug safety requirements and may be a cause of both adverse effects and drug interactions. Data on interactions between herbal remedies and conventional antirheumatic medication is scarce. Reasons include a perception that herbal remedies are safe, a lack of reporting by patients and healthcare professionals and a lack of knowledge about the pharmacology and composition of herbal remedies, as well as adulteration. Interactions are likely between herbal remedies with antiplatelet or nephrotoxic effects and NSAIDs, hepatotoxic herbal remedies and disease-modifying antirheumatic medication, and between St. John's Wort and cyclosporin. PMID- 16255657 TI - Alefacept: a safety profile. AB - Alefacept is a selective immunomodulating, antipsoriatic drug that blocks the LFA 3/CD2 interaction necessary for the activation and proliferation of memory effector T cells by binding to CD2 expressed on the T cell surface. Because the CD4+ count is reduced by alefacept, it is recommended that this count be monitored on a regular basis to ensure that it does not drop below 250 cells/mul. Few side effects have been related to the use of alefacept that differ from placebo even when CD4+ counts drop below 250 cells/microl. The side effects that have been reported are minor and include: headache, nasopharyngitis, rhinitis, influenza, upper respiratory tract infections, pruritus, arthralgias, fatigue, nausea, accidental injury and increases in liver enzymes. Serious infections and malignancies do not appear linked to the use of alefacept. The percentage of patients who developed antibodies against alefacept is very low. Alefacept is a very safe biological therapy for moderate-to-severe chronic plaque psoriasis with few side effects reported. The utility of checking CD4 counts while administering alefacept for 12 weeks appears minimal. PMID- 16255658 TI - Safety and adverse event profile of duloxetine. AB - Duloxetine is the first relatively balanced serotonin and noradrenaline re-uptake inhibitor to be widely available for three indications including: major depressive disorder, peripheral diabetic neuropathic pain and female stress urinary incontinence, although it is not currently approved for all indications in all countries. Generally, duloxetine is safe and well-tolerated across indications, with few reported serious side effects. Common adverse events are consistent with the pharmacology of the molecule and are mainly referable to the gastrointestinal and the nervous systems. The studied dose range is up to 400 mg/day (administered 200 mg b.i.d) but the maximum dose approved for marketing is 120 mg/day (administered 60 mg b.i.d). Duloxetine is eliminated (half-life = 12.1 hours) primarily in the urine after extensive hepatic metabolism by multiple oxidative pathways, methylation and conjugation. Duloxetine would not be expected to cause clinically significant inhibition of the metabolic clearance of drugs metabolised by P450 (CYP)3A, (CYP)1A2, (CYP)2C9, or (CYP)2C19, but would be expected to cause some inhibition of CYP 2D6. Duloxetine should not be used in combination CYP 1A2 inhibitors or nonselective, irreversible monoamine oxidase inhibitors. The purpose of this review is to provide an overview of some of the most important information related to safety and tolerability of duloxetine. PMID- 16255659 TI - Bupropion: risks and benefits. AB - Cigarette smoking represents one of the most preventable causes of death worldwide. However, success rates for stopping smoking are disappointingly low and are associated with high relapse rates. There is a need for a successful form of smoking cessation therapy. Bupropion is an effective therapy for smoking cessation and is recommended as first-line treatment in both US and UK guidelines. Its mechanism of action in smoking cessation is unclear, although it is thought that dopaminergic pathways are involved in the 'reward' circuit of drug dependence. Seizures are an important adverse effect of bupropion and care is needed when used in other conditions or with other medication that can lower the seizure threshold. Bupropion has been shown to be a cost-effective therapy for smoking cessation. PMID- 16255660 TI - Celecoxib and cardiovascular risks. AB - In a very short time, COX-2 enzyme inhibitors have gone from the darlings to the pariahs of the pharmaceutical industry. These drugs were developed based on the hypothesis whereby selective inhibition of the COX enzyme would lead to reduction in pain and inflammation without associated gastrointestinal and bleeding risks. However, in September 2004, rofecoxib was voluntarily removed from the market for increased cardiovascular risk and in April 2005, valdecoxib was also withdrawn, at least in part, due to excess cardiovascular risk. Celecoxib was the first COX 2 inhibitor introduced and the only remaining one on the US market. There is consequently a justified concern that cardiovascular toxicity is a class effect of all COX-2 inhibitors. This article systematically reviews the evidence surrounding COX-2 inhibitors and cardiovascular risk. Although the evidence suggests a fairly consistent cardiovascular risk with rofecoxib, the evidence for cardiovascular risk with celecoxib is more equivocal. Although isolated studies have suggested some cardiovascular risk for celecoxib, the totality of the evidence suggests that any risk is likely to be small and comparable to traditional NSAIDs. The cardiovascular risks of COX-2 inhibitors appear heterogeneous, influenced not only by the drug class, but also individual drug, dosage and patient characteristics. Specific modifying factors of the cardiovascular risk of COX-2 inhibitors including dose, concomitant drugs, individual cardiac and genetic risk profiles, will require further study. PMID- 16255661 TI - Cardiovascular disease and toxicities related to HIV infection and its therapies. AB - Cardiovascular manifestations of HIV vary according to disease stage, treatment regimen and geographical location. Common cardiac complications of HIV disease in patients off highly active antiretroviral therapy (HAART) include dilated cardiomyopathy, myocarditis, pericardial effusion, endocarditis, pulmonary hypertension and non-antiretroviral drug-related cardiotoxicity. However, with the introduction of HAART that has substantially modified the course of HIV disease by lengthening survival, additional cardiovascular consequences are a result of the metabolic syndrome with a propensity toward hyperlipidaemia and atherosclerotic heart disease. Because most of the world's HIV-infected patients have not been treated with HAART, the principal HIV-associated cardiovascular manifestations of patients off HAART are reviewed and new knowledge about the prevalence, pathogenesis and treatment in the HAART era are emphasised in this review. Exercise, a nonpharmacological approach to treating HAART-associated metabolic syndrome, is also discussed. PMID- 16255662 TI - The safety and efficacy of the use of vasopressin in sepsis and septic shock. AB - Sepsis remains a significant problem and cause of morbidity and mortality in intensive care. Vasopressin infusions are currently used as rescue therapy for the treatment of vasodilatory, catecholamine-resistant septic shock. At present, there are no large randomised, controlled trials in the literature investigating vasopressin in this role, although two such studies are currently ongoing in Canada. This review outlines the pathophysiology of sepsis and that of vasopressin in sepsis and reviews the available evidence for the use of vasopressin in sepsis and septic shock. A review of the safety data for vasopressin in this indication is included. Recommendations for the use of vasopressin in septic shock, along with suggestions for the direction of further work in the field are presented. PMID- 16255663 TI - Elderly patients with advanced colorectal cancer: which therapy is the safest? AB - Patients > 65 years of age are the fastest growing segment of the cancer population. It is estimated that within 20 years, > 75% of cases and 85% of deaths from colorectal cancer (CRC) will be in this setting. Concerns about cancer treatment in the elderly relate to comorbidities, which increase proportionally with age, physiological changes associated with ageing that may influence drug metabolism and toxicity, and diminishing life expectancy, which particularly impacts decisions surrounding the benefits of adjuvant therapies. Over the last 10 years, significant improvements in the treatment of advanced CRC with combination therapy have been made. The randomised trials that have defined these improvements did not exclude elderly patients; however, the median age of patients in these trials has generally been approximately 60 years. Thus, it appears that some degree of selection is involved with younger and presumably fitter patients being the subjects in most of the pivotal trials. The availability of new molecularly targeted agents and newly improved existing agents has expanded the range of treatment options available. This variety gives greater flexibility in dealing with different subsets of patients, such as the elderly. However, some fit elderly patients seem to tolerate combination therapy reasonably well, whereas studies on unfit elderly subjects are needed. PMID- 16255664 TI - Safety profile of platinum-based chemotherapy in the treatment of advanced non small cell lung cancer in elderly patients. AB - Non-small cell lung cancer (NSCLC) may be considered typical of advanced age. More than 50% of NSCLC patients are diagnosed at > 65 years of age and approximately one-third of all patients are > 70 years of age. Elderly patients tolerate chemotherapy poorly compared with their younger counterpart because of the progressive reduction of organ function and comorbidities related to age. For this reason, these patients are often not considered eligible for aggressive platinum-based chemotherapy, the standard medical treatment for advanced NSCLC. In clinical practice, single-agent chemotherapy should remain the standard treatment. Feasibility of platinum-based chemotherapy remains an open issue and has to be proven prospectively. Moreover, a multidimensional geriatric assessment for individualised treatment choice in NSCLC elderly patients is mandatory. This review focuses on the currently-available evidences for the treatment of elderly patients affected by advanced NSCLC with regards to the role and safety of platinum-based chemotherapy. PMID- 16255665 TI - Safety and efficacy of growth hormone replacement therapy in adults. AB - Limitless supplies of recombinant human growth hormone (GH) have been available for the last 20 years. During that period, studies have characterised the effects of GH deficiency in adults and the benefits of GH replacement therapy. Areas of greatest impairment and benefit are quality of life, skeletal health and cardiovascular risk factors including the serum lipid profile and body composition. By optimising GH replacement therapy at various stages of adult life, it is hoped that it will prevent the development of osteoporosis and reduce the mortality and morbidity associated with hypopituitarism. However, the primary indication for GH therapy in adults in England and Wales is quality of life. The benefits of GH treatment are sustained over several years, and long-term surveillance of patients continues. PMID- 16255666 TI - Safety of obesity drugs. AB - The safety of obesity drugs has historically been poor. This and the stigmatisation of obesity in society ensured that a higher standard of safety for obesity drugs must be met. The authors review the safety disasters of obesity drugs that were withdrawn. The authors then review the safety of presently available drugs--benzphetamine, phendimetrazine, diethylpropion, phentermine, sibutramine and orlistat. The safety of rimonabant, a drug with a pending new drug application that has an independent effect on metabolic syndrome, is also reviewed. The authors compare the stage of obesity drug development to that of hypertension in the 1950s. As new and safer drugs with more downstream mechanisms are developed that have independent effects on the cardiovascular risks associated with obesity, third party reimbursement for obesity medicine is likely to improve. This may lead to obesity being treated like hypertension and other chronic diseases with long-term medication. With improved technological tools, the authors believe this process will be more rapid for obesity than it was for hypertension. PMID- 16255667 TI - Drug-induced diabetes mellitus. AB - AIMS: To review the medications that influence glucose metabolism with a focus on hypertensive, transplant and HIV-infected patient populations. METHODS: Literature obtained from a MEDLINE search from 1970 to present, including studies published in the English language. The search strategy linked drugs, hyperglycaemia and diabetes mellitus, HIV, transplantation, hypertension and psychiatric patients. RESULTS: Many common therapeutic agents influence glucose metabolism. Multiple mechanisms of action on glucose metabolism exist through pancreatic, hepatic and peripheral effects. The prevalence of hyperglycaemia was higher with the use of thiazide diuretic, beta-blocker, calcineurin, protease inhibitors and atypical antipsychotic drugs. CONCLUSIONS: Patients treated with those drugs appear to be at increased risk for developing diabetes. It is prudent to monitor plasma glucose values when it is not possible to avoid prescription of medication with known effects on carbohydrate metabolism. PMID- 16255668 TI - Atypical antipsychotic drugs, diabetes and ethnicity. AB - There are 17 million people affected by diabetes in the US. It is a syndrome consisting of metabolic abnormalities, microvascular and macrovascular disease leading to cardiac, renal and neurological abnormalities. Obesity is the most common public health problem in developed nations. Diabetes and obesity-related illnesses are common in ethnic minorities such as African-Americans, Hispanics and Asians related to both genetics and lifestyle patterns. In all ethnic minorities in the US, an increase in Type 2 diabetes has been observed. However, the Asian group experienced the highest rate of increase in prevalence between the years 1990 and 1998. The changing ethnic composition of the US population may contribute significantly to the worsening of the diabetes epidemic in this country. Atypical antipsychotic drugs can induce diabetes, as well as obesity. All atypical antipsychotic drugs can produce diabetes, but drugs such as olanzapine and clozapine have been known to produce diabetes more often than other drugs. As ethnic minority patients including Asians, Hispanics and African Americans are predisposed to develop diabetes, antipsychotics become a burden by precipitating diabetes. Such a situation poses a problem in treating ethnic minority psychiatric patients. In clinical situations, close monitoring is necessary to prevent metabolic side effects of these drugs. PMID- 16255669 TI - The potential dangers of treating head injury patients with corticosteroids. AB - In the past, corticosteroids were given to head-injured patients in order to prevent secondary brain damage, even if clinical trials had been inconclusive and potential risks of complications were of concern. Recently, CRASH, a large, multi centre study on short-term, high-dose corticosteroid treatment in head trauma, was interrupted after enrolling > 10,000 patients because corticosteroid treatment was associated with significantly higher mortality within two weeks. Participating clinicians were not requested to judge the causes of death, but rates of infections and gastrointestinal haemorrhages did not differ between treated patients and controls. Other potential corticosteroid complications include metabolic derangements (particularly hyperglycaemia), adrenal insufficiency and critical illness myopathy. Furthermore, experimental data suggest that corticosteroids may have some harmful effects on neural tissue. In this review, the potential risks of treating head-injured patients with corticosteroids are examined. PMID- 16255670 TI - Probiotics under the regulatory microscope. AB - This review examines current knowledge regarding the safety of probiotic bacteria in man. Tighter and more comprehensive standards and regulations will be developed as probiotic therapy moves from a limited number of products used in the food industry, into more defined therapeutic categories and more complex organisms. A new framework considering probiotics as nonspecific promoters of mucosal immunity, defines probiotic characteristics and the clinical circumstances in which it is used. For example, those with immune deficiency taking a high dose of viable bacteria may have an increased risk. A wider range of bacteria is now being used, sometimes in territories other than the gut mucosa. The question of competition with multiple isolates must be addressed, as does the use of nonselected faecal isolates. Transfer of antibiotic resistance with probiotics acting as a 'shuttle' needs clarification. These issues are addressed and reviewed as probiotics evolve into a new therapeutic arena. PMID- 16255671 TI - The potential adverse effects of habitual use of Catha edulis (khat). AB - The habit of Catha edulis Forsk (khat) chewing has prevailed for centuries among populations in the horn of Africa and the Arabian peninsula. Fresh leaves of Catha edulis are customarily chewed to attain a state of stimulation. The fact that cathinone has a closer structural similarity with amphetamine, and both share common pharmacodynamic features, led to the conclusion that cathinone is the most important active ingredient of Catha edulis, which causes the major pharmacological effects. Problems associated with repeated consumption of Catha edulis leaves are becoming evident. Literature surveys and clinical diagnostic studies revealed an association with prevalence of psychiatric morbidity. Repeated intermittent oral administration of Catha edulis to laboratory animals lead to development and expression of locomotor sensitisation. Catha edulis also leads to several peripheral effects that include increases in blood pressure and has been associated with the increased incidence of acute coronary vasospasm and myocardial infarction. Catha edulis also accounts for a number of gastrointestinal tract problems, oesophagitis, gastritis and delay intestinal absorption, as well as the development of oral keratotic white lesions at the site of chewing. Administration of Catha edulis extracts showed a deranged systemic capacity to handle oxidative radicals and induces cytotoxic effects in cells of liver and kidney, as well as induction of cell death in various human leukaemia cell lines and in peripheral human blood leukocytes. PMID- 16255672 TI - Therapeutic applications of carbon monoxide-releasing molecules. AB - Carbon monoxide (CO), which is formed in mammalian cells through the oxidation of haem by the enzyme haem oxygenase, actively participates in the regulation of key intracellular functions. Emerging evidence reveals that an increased generation of haem oxygenase-derived CO plays a critical role in the resolution of inflammatory processes and alleviation of cardiovascular disorders. The authors have identified a novel class of substances, CO-releasing molecules (CO-RMs), which are capable of exerting a variety of pharmacological activities via the liberation of controlled amounts of CO in biological systems. A wide range of CO carriers containing manganese (CORM-1), ruthenium (CORM-2 and -3), boron (CORM A1) and iron (CORM-F3) are currently being investigated to tailor therapeutic approaches for the prevention of vascular dysfunction, inflammation, tissue ischaemia and organ rejection. PMID- 16255673 TI - Endothelial function and its assessment. AB - Endothelial dysfunction is a characteristic aspect of most of the conditions associated with atherosclerosis and is commonly found as an early feature in atherothrombotic vascular disease. An appreciation of the underlying mechanisms of endothelial function, as well as dysfunction, is essential as this has critical influence on the different methods in the assessment of endothelial function and effects of various treatments on its quantification. Furthermore, endothelial dysfunction is recognised as a type of 'target organ damage' in common cardiovascular conditions (e.g., hypertension) and the area is of increasing interest for new drug development, as therapies that modulate the endothelium will have added advantages; thus, for the development of new/experimental drugs, an awareness of ways to assess the endothelium is necessary. In this review, an overview of different methods including biochemical markers, and invasive and non-invasive tools, to determine endothelial function is presented as well as their clinical relevance. Furthermore, the effects of various treatments on endothelial dysfunction and their underlying mechanisms are elucidated. PMID- 16255674 TI - Clinical status of agents being developed for leishmaniasis. AB - Leishmaniasis, which exists in both visceral and cutaneous forms, is currently treated with intramuscular antimony or intravenous amphotericin B. The primary unmet need is for oral therapy. Of the several drugs in clinical development, miltefosine is unique in being an oral agent with efficacy against both forms of the disease. Sitamaquine is an oral agent with substantial but not sufficient efficacy against visceral disease. Oral fluconazole has been shown to be more effective than placebo in one instance: for Leishmania major cutaneous disease from Saudi Arabia. Paromomycin is in widespread trial. Topical paromomycin formulations are being tested for cutaneous disease, and intramuscular paromomycin is in Phase III trial for Indian visceral disease. The most likely replacements for present therapy are oral miltefosine for many of the visceral and cutaneous syndromes, intramuscular paromomycin for visceral disease and topical paromomycin for some forms of cutaneous disease. PMID- 16255675 TI - Nitric oxide-modulating agents for gastrointestinal disorders. AB - Almost 20 years after the identification of the biological role of nitric oxide (NO), the full therapeutic potential of novel agents that mimic the activity of NO or interfere with its synthesis has yet to be realised for utilities involving the gastrointestinal tract. New utilities for classical NO donors, which were used as vasodilators for decades, in the treatment of motility disorders have been explored and a product for treating anal fissure was recently launched. New classes of compounds incorporating a NO-donating moiety into standard non steroidal anti-inflammatory drugs, the NO-non-steroidal anti-inflammatory drugs (NO-NSAIDs) or COX-inhibiting nitric oxide donors (CINODs) have also been developed. These have been shown to exhibit reduced gastrointestinal injury in experimental models, and reports on their efficacy and safety in Phase I and II studies are now available. Modulation of the inducible NO synthase isoform that generates excessive NO that can lead to subsequent cytotoxic moieties, such as peroxynitrite, may have therapeutic possibilities in a range of inflammatory diseases of the gut. Likewise, agents that promote the decomposition of peroxynitrite or removal of its other component, superoxide, may also prove to be of use. Further targets for pharmaceutical exploitation are likely to come from both genomic and molecular insights into the processes that regulate the NO system. PMID- 16255676 TI - Benign prostatic hyperplasia: an insight into current investigational medical therapies. AB - Benign prostatic hyperplasia (BPH) is a leading disorder of the elderly male population that is characterised by a progressive enlargement of prostatic tissue, resulting in obstruction of the proximal urethra and causing urinary flow disturbances. The pathophysiology of BPH associated with lower urinary tract symptoms is characterised by increased adrenergic tone (dynamic component) leading to smooth muscle contraction and prostatic overgrowth due to androgenic stimulation (static component); therefore, the therapeutic armamentarium of BPH can be broadly divided into antiadrenergic and antiandrogenic approaches. alpha1 Adrenoceptor antagonists and 5alpha-reductase inhibitors are well-established representatives of the two categories, respectively. Other antiandrogenic approaches involve gonadotropin-releasing hormone agonists and antagonists for the treatment of prostate hyperplasia. Apart from these approaches, new approaches with novel targets are emerging. The advent of new therapies is, however, more oriented towards the static component. These involve metabolic factors (hexokinase inhibitor), growth factors (vitamin D3 analogues), oxytocin antagonists and gonadotropin-releasing hormone Gi agonist-based therapies. Gene therapy and photodynamic therapies are other emerging therapies for relieving symptoms in BPH patients. With the initial success of upcoming targets, the unmet need to develop an efficacious and relatively safe therapeutic modality is discussed. Nevertheless, their long-term safety and efficacy needs to be evaluated in large-scale clinical trials. The future also belongs to combination therapies to combat both dynamic and static disease components and for extended indications such as micturition disorder and non-bacterial prostatitis. PMID- 16255677 TI - Agents targeting c-Jun N-terminal kinase pathway as potential neuroprotectants. AB - c-Jun N-terminal kinase (JNK) plays an integral role in neuronal death in multiple cell lines following a wide variety of stimuli and in a number of physiological functions that may be involved in human disease, including CNS diseases. In the past decades, many researchers in this field have found and reinforced the concept that prolonged activation of JNK signalling can induce neuronal cell death by both a transcriptional induction of death-promoting genes and modulation of the mitochondrial apoptosis pathways. Data are emerging to extend the understanding of the JNK signalling and confirm the possibility that targeting JNK signalling may offer an effective therapy for pathological conditions in the near future. This review will focus on the pro-apoptotic role of JNK signalling and updated pharmacological inhibitors of this pathway. PMID- 16255678 TI - Protease inhibitors as potential disease-modifying therapeutics for Alzheimer's disease. AB - The current lack of an effective treatment for Alzheimer's disease (AD) has fuelled an intense search for novel therapies for this neurodegenerative condition. Aberrant production or decreased clearance of amyloid-beta peptides is widely accepted to be causative for AD. Amyloid-beta peptides are produced by sequential processing of the beta-amyloid precursor protein by the two aspartyl type proteases beta-secretase and gamma-secretase. Because proteases are generally classified as druggable, these secretases are a centre of attraction for various drug discovery efforts. Although a large number of specific drug-like gamma-secretase inhibitors have been discovered, progress towards the clinic has been slowed by the broad substrate specificity of this unusual intramembrane cleaving enzyme. In particular, the Notch receptor depends on gamma-secretase for its signalling function and, thus, gamma-secretase inhibition produces distinct phenotypes related to a disturbance of this pathway in preclinical animal models. The main task now is to define the therapeutic window in man between desired central efficacy and Notch-related side effects. In contrast, most studies with knockout animals have indicated that beta-secretase inhibition may have minimal adverse effects; however, the properties of the active site of this enzyme make it difficult to find small-molecule inhibitors that bind with high affinity. In most instances, inhibitors are large and peptidic in nature and, therefore, unsuitable as drug candidates. Thus, there are many issues associated with the development of protease inhibitors for AD that must be addressed before they can be used to test the 'amyloid cascade hypothesis' in the clinic. The outcomes of such trials will provide new directions to the scientific community and hopefully new treatment options for AD patients. PMID- 16255679 TI - Immunomodulatory drugs in multiple myeloma. AB - Immunomodulatory drugs (IMiDs) are thalidomide analogues that retain the direct anticancer cytotoxic and immunological activity of their parent compound, but with a different toxicity profile. In vitro studies show that IMiDs have a more potent antitumour effect than thalidomide on multiple myeloma (MM) cell lines. This activity is mediated by multiple mechanisms: direct antiproliferative effect; inhibition of angiogenesis due to reduced IL-6 and vascular endothelial growth factor secretion; inhibition of cytokines production, especially TNF alpha; and stimulation of T-cell activity. Two IMiDs, CC-5013 and CC-4047, have been tested in clinical trials in MM patients with progressive or refractory disease, and one trial is ongoing in newly diagnosed MM patients. Observed toxicities include thrombocytopoenia, neutropoenia and cardiovascular events, but no significant neurotoxicity has been reported. Partial responses (> or = 50% reduction in M-protein) ranged from 20 to 71% in different studies depending on the pretreatment status of the patients. The combination of IMiDs with dexamethasone may be beneficial. PMID- 16255681 TI - MC-1 (pyridoxal 5'-phosphate): novel therapeutic applications to reduce ischaemic injury. AB - Despite the overall efficacy of mechanical reperfusion therapies, such as percutaneous coronary intervention and coronary artery bypass graft surgery, in reducing the morbidity and mortality that is associated with acute ischaemic syndromes, many of the treated patients develop ischaemia-reperfusion injury due to impaired microvascular integrity, embolisation of atherothrombotic debris and/or disrupted end-organ metabolism. MC-1 is an investigational drug from Medicure, Inc. In preclinical models of ischaemia and ischaemia-reperfusion injury, treatment with MC-1 has demonstrated significant cardio- and neuroprotective effects. Although the pharmacological activity of MC-1 may involve multiple mechanisms, research suggests that at least part of the protective effect may be mediated through its actions on purinergic receptors. Early clinical experience with MC-1 also appears to be promising: in a recent Phase II evaluation, treatment with MC-1 was associated with a statistically significant reduction in periprocedural infarct size (as measured by area under the curve creatine kinase-myocardial band) among high-risk patients undergoing elective percutaneous coronary intervention. Based on these findings, larger, randomised trials to confirm the safety and efficacy of MC-1 in the setting of coronary artery revascularisation with coronary artery bypass graft, acute coronary syndromes and stroke are ongoing or in development. These forthcoming evaluations should clarify the safety and efficacy of MC-1 and improve the understanding regarding its potential therapeutic role in a variety of clinical settings and indications. PMID- 16255680 TI - Investigations on gossypol: past and present developments. AB - Gossypol has received significant attention as a result of its potential therapeutic application as a male antifertility agent. Furthermore, recent research examining the biological activity of gossypol has revealed a number of other promising lines of enquiry. These have focused on the antitumour, antiviral and antioxidant actions of the compound in various disease states. This article provides an overview of the studies on the biological activity of gossypol, with particular attention paid to the mechanisms of its activity and its prospect as a medicinal product. PMID- 16255684 TI - Retinal degeneration: challenge and opportunity. PMID- 16255682 TI - Pirfenidone for the treatment of idiopathic pulmonary fibrosis: therapeutic potential prompts further investigation. PMID- 16255685 TI - Sifting the available evidence on age-related macular degeneration. PMID- 16255686 TI - Anatomy and development of the macula: specialisation and the vulnerability to macular degeneration. AB - The central retina in primates is adapted for high acuity vision. The most significant adaptations to neural retina in this respect are: 1. The very high density of cone photoreceptors on the visual axis; 2. The dominance of Midget pathways arising from these cones and 3. The diminishment of retinal blood supply in the macula, and its absence on the visual axis. Restricted blood supply to the part of the retina that has the highest density of neural elements is paradoxical. Inhibition of vascular growth and proliferation is evident during foetal life and results in metabolic stress in ganglion cells and Muller cells, which is resolved during formation of the foveal depression. In this review we argue that at the macula stressed retinal neurons adapt during development to a limited blood supply from the choriocapillaris, which supplies little in excess of metabolic demand of the neural retina under normal conditions. We argue also that while adaptation of the choriocapillaris underlying the foveal region may initially augment the local supply of oxygen and nutrients by diffusion, in the long term these adaptations make the region more vulnerable to age-related changes, including the accumulation of insoluble material in Bruch's membrane and beneath the retinal pigment epithelium. These changes eventually impact on delivery of oxygen and nutrients to the RPE and outer neural retina because of reduced flow in the choriocapillaris and the increasing barriers to effective diffusion. Both the inflammatory response and the sequelae of oxidative stress are predictable outcomes in this scenario. PMID- 16255687 TI - Retinal remodelling. AB - Retinal degenerative diseases that progress through loss of photoreceptors initiate a sequence of events that culminates in negative remodelling of the retina. Initially, photoreceptor loss ablates glutamatergic signalling to the neural retina and eliminates coordinate Ca++-coupled homeostatic signalling. Retinal neurons react to this loss of glutamatergic input through retinal rewiring and migration of neurons throughout the axis of the retina. All diseases that kill photoreceptors trigger retinal remodelling as the final common pathway and cell death is a common feature. Retinal remodelling resembles CNS pathologic remodelling and constitutes a major challenge to all rescue strategies. PMID- 16255688 TI - Assessment of age-related maculopathy using subjective vision tests. AB - This paper reviews non-standard, clinical vision tests that may be used to detect the earliest visual loss in age-related maculopathy (ARM), before fundus changes are detected. We recommend a clinical test battery for all patients aged 60 years and older, comprising low luminance/low contrast (SKILL) VA or low contrast VA, desaturated D-15 colour vision assessment, flicker perimetry, glare recovery and dark adaptation if possible, together with conventional assessments of case history, ophthalmoscopy and high contrast visual acuity (VA) for the detection and diagnosis of ARM. Reading rate is also discussed as a potential indicator of early visual loss. For monitoring the progressive visual loss in age-related macular degeneration (AMD) and determining the requirements for optometric vision rehabilitation, we recommend more conventional clinical vision tests of distance and near visual acuity, reading rate, the effects of varying illumination and a functional central visual field assessment. PMID- 16255689 TI - Objective functional assessment of age-related maculopathy: a special application for the multifocal electroretinogram. AB - This paper gives a brief review of methods that assess objectively function in age-related maculopathy (ARM) with emphasis on a newer method, the multifocal electroretinogram (mfERG). In contrast to other electrophysiological tests, such as the full-field and focal electroretinogram (ERG) or the electro-oculogram (EOG), which measure summed responses from various cells from larger areas of the retina, the multifocal electroretinogram maps function locally with a resolution as small as four degrees within the central 30 degrees. By using different paradigms it can measure local cone- and rod-mediated functional impairment at early and late stages of ARM. This improved mapping and higher resolution of the posterior pole compared to other objective methods might lead to earlier detection of ARM. Its usefulness has been demonstrated in documenting the effects of treatment after established laser treatments, such as photodynamic therapy (PDT) and in documenting function after retinal pigment epithelial transplantation, a possible future treatment in late neovascular ARM. PMID- 16255690 TI - Visual function: the problem with eccentricity. AB - Age-related macular degeneration (AMD) is the leading cause of blindness in developed countries. With an ageing population, the prevalence of such a condition has resulted in a large proportion of the population relying on peripheral vision to undertake activities of daily living. Peripheral vision is not a scaled-down version of the fovea, simply requiring larger print or increased contrast for detection of objects or reading text. Even when print size is scaled and eye movements are minimised, the peripheral retina cannot perform at the level of the foveal region. Understanding how and why reading performance is limited as a function of eccentricity has important implications for how we approach rehabilitation of patients with central visual loss. This brief review of the extensive literature on reading with peripheral vision and the research aimed at better reading rehabilitation for low vision patients focuses on why many of the problems associated with the reduced reading capability of peripheral vision cannot be completely solved with magnification, reducing eye movements or modifying print. PMID- 16255691 TI - Treatment of age-related macular degeneration. AB - Age-related macular degeneration (AMD) is the greatest cause of legal blindness in the western world. Established treatments include argon laser photocoagulation of extrafoveal choroidal neovascularisation (CNV) and photodynamic therapy of selected sub-foveal CNV. Newer approaches are targeting the angiogenic pathway in CNV development. Currently, other treatment modalities, such as radiotherapy and transpupillary thermotherapy do not have a clear role to play. Surgical options are experimental and only available in some centres for selected patients. Prevention of AMD remains elusive. Dietary supplements may have a role, while statins and prophylactic laser photocoagulation of drusen remain experimental. This paper explains the principles behind these approaches. PMID- 16255692 TI - Retinitis pigmentosa: visual function and multidisciplinary management. AB - Retinitis pigmentosa (RP) is a leading cause of blindness and visual disability in younger people. Optometrists have a major role in detecting RP and in reducing the visual disability associated with RP. This review summarises the literature relating to visual function in people with RP, with particular attention given to night-blindness, visual acuity decrease and visual field contraction. The range of low vision aids available for people with RP is reviewed and suggestions given on aids that have been found to be most successful. Most importantly, this review overviews the range of services available to people with RP and emphasises how optometrists need to work with a network of professionals to ensure the best possible visual outcomes for people with RP. Particular mention is made of current findings relating to orientation and mobility training, driving, sensory substitution and adaptive technology. The modern optometrist needs to be aware of the multiple needs of people with RP and have the ability to link them with the professionals best able to help them. PMID- 16255693 TI - Take supplements with a grain of salt. PMID- 16255697 TI - Change through continuity: a quiet revolution in primary health care in New Zealand. PMID- 16255695 TI - No longer the only colour defective cricketer. PMID- 16255698 TI - Markets in health care: taking a tiger by the tail? PMID- 16255699 TI - Slaves to economists? A Canadian's view of the Australian health care system. PMID- 16255700 TI - From council to commission: building on a solid foundation for safety and quality. PMID- 16255701 TI - The Australia and New Zealand horizon scanning network. PMID- 16255702 TI - Information sharing in health care: a patient's perspective. PMID- 16255703 TI - Commentary: the patient's memory stick may complement electronic health records. PMID- 16255704 TI - Managing quality in cancer services: why improvement isn't easy. AB - Optimising the quality of care is an imperative for health services worldwide, including in Australia. Recognition that poor quality often has its roots in system failures is beginning to shift strategies for improvement to the systems of care, although the tendency remains to focus on eliminating the practice variations of individual clinicians. In those instances where systems improvement is addressed, strategies tend to be generic and technical, and often unrelated to the context in which they are applied. This paper reports an interim evaluation of a quality management program in cancer services implemented in a Sydney metropolitan teaching hospital dispersed across multiple campuses. The paper aims to inform the debate on quality improvement by reporting not only on what was achieved, but why change seems to be so hard. We found that organisational and social factors that influence the quality of health services were not sufficiently addressed, compared with technical factors. We conclude that service quality needs to be repositioned as an organisational goal, and implemented via a structured process that addresses organisational and social factors, as well as technical factors. PMID- 16255705 TI - Antibiotic guidelines in NSW hospitals. PMID- 16255706 TI - Implementing root cause analysis in an area health service: views of the participants. AB - PURPOSE: This study identifies the attitudes of participants in the root cause analysis (RCA) process and barriers to it's implementation within one New South Wales area health service. METHOD: Employees and consumer representatives of the former South Western Sydney Area Health Service who participated in an RCA as either a team member or a team leader between December 2002 and October 2003 completed a self-administered survey. RESULTS: Thirty seven of 39 eligible participants completed the survey (response rate 95%). The respondents identified formulation of causal statements, ensuring the causal statements met the "rules of causality" outlined by New South Wales Health, and arranging times for interviews as most difficult. Team leader respondents (n = 7) ranked keeping the team focused, organising the first meeting within 7 days of the incident, and completing the RCA in three 2-hour meetings as barriers to the process. CONCLUSIONS: Training was valued by participants, however greater emphasis on the development of causal statements could be beneficial. Team leaders expressed difficulty in keeping the team focused and meeting the stipulated RCA timeframes, suggesting that additional support for RCA participants may be warranted. PMID- 16255707 TI - The first eighteen months of a paediatric ambulatory and community service. AB - Hospital admission is distressing and traumatic for children as they are separated from their families and home. Internationally, and in Australia, alternative models of health care are being developed to meet the needs of children and their families. We describe the first eighteen months of the establishment of a paediatric ambulatory and community service in a district health service in New South Wales. Key outcomes include: increased referral to the service from acute hospital and primary care services; parental satisfaction and saved hospital bed-days. Lessons learnt in the setting up of this service include the need for proactive engagement of consumers and stakeholders; clear definition of roles and responsibilities; and measurable and reasonable performance indicators. PMID- 16255708 TI - A central role for the health call centre. PMID- 16255709 TI - Assessing patient satisfaction: implications for South Australian public hospitals. AB - This paper reports on the results from 2620 South Australians who participated in the 2003 Patient Evaluation of Hospital Services. Patients were found to be generally satisfied with the care, services and amenities provided, with a statewide overall score of 86.3. Satisfaction was lowest in the patients' assessment of their involvement in their own care and treatment. Three demographic factors (younger age, female sex or tertiary education) predicted lower levels of satisfaction in the multivariate analysis, whereas living with others, non-emergency admission or admission to smaller hospitals were found to predict higher satisfaction. Despite administrative and organisational difficulties, and limited current evidence of increased quality or satisfaction, it is considered important to continue satisfaction research with the goal of encouraging the development of action plans for improvement of care, services and amenities. PMID- 16255710 TI - How do rural consumers contribute to quality assurance of health services? AB - OBJECTIVE: To investigate the reasons for complaint or non-complaint by rural consumers of health services. DESIGN: Qualitative study using focus group discussion of hypothetical scenarios. SETTING: Selected rural communities in the Loddon-Mallee region of north-western Victoria. PARTICIPANTS: Sixty volunteer participants in eight focus groups recruited through advertising. MAIN OUTCOME MEASURE: Issues and themes concerning circumstances leading to, and factors inhibiting, complaints about a health service and awareness of complaints mechanisms. RESULTS: Compared with residents of larger towns, those of small communities were more likely to report they would complain to the local provider, whereas those in larger towns were more likely to mention Hospital Boards or the Commissioner. Deterrents to making complaints included the lack of services, scepticism about the role of complaints in bringing about change and an attitude that it was more appropriate to try to fix the problem than complain about it. Lack of awareness of appropriate complaint mechanisms which feed into quality assurance processes was also identified. CONCLUSIONS: Previously documented lower complaint rates from rural consumers can not be taken to mean greater satisfaction with health services. PMID- 16255711 TI - Identification of Australians from culturally and linguistically diverse backgrounds in national health data collections. AB - In multicultural Australia, comprehensive and up-to-date information on ethnicity and health is essential to guide policy and service development in the health sector. Data collected for purposes other than research are a potentially important source of information. This study explored the extent to which indicators of cultural and linguistic diversity are currently included in national health and welfare service data collections, and the data standards employed. We identified and reviewed 44 relevant bodies of work: 7 national data dictionaries, 15 national data sets, 10 national health data collections and 12 national surveys. Each of the large data dictionaries (health, community services and housing assistance) contained several ethnicity-related variables. Immigrant Australians were identified (usually by country of birth, sometimes by language, and occasionally by period of residence or year of arrival) in all the major national health and community data sets, health data collections and surveys. Australian Bureau of Statistics standards and classifications relating to cultural and linguistic diversity were widely used. Researchers, health policy makers and planners should fully exploit these secondary data sources, as well as undertaking or commissioning primary research. PMID- 16255712 TI - Enhancing evidence-based practice in population health: staff views, barriers and strategies for change. AB - STUDY OBJECTIVE: To determine barriers and enablers for evidence-based practice (EBP) in population health and potential strategies for change. DESIGN: Self administered survey of 104 professional staff (response rate, 73%) in the Division of Population Health, South Western Sydney Area Health Service in NSW serving a disadvantaged urban population. MAIN RESULTS: Most respondents (80%) "strongly agreed" or "agreed" that EBP would improve the effectiveness of their efforts in a disadvantaged region. However, more than half of respondents (56%) "strongly agreed" or "agreed" that there is lack of evidence for interventions in population health. Eighty two per cent of respondents "strongly agreed" or "agreed" that training in EBP is important for all population health workers. Those who used evidence also needed a greater capacity to discriminate "good" from "bad" research (85% in agreement). Contradictory policy was cited by one third of respondents as acting against EBP. PMID- 16255713 TI - Using pilot studies to inform health services. AB - A pilot study was conducted at the John Hunter Hospital, Newcastle, Australia in 1998-99 to inform a randomised controlled trial (RCT) for a cardiac rehabilitation intervention for patients with congestive heart failure (CHF). Although the RCT did not proceed, the pilot study results raised a number of issues. In this paper, the pilot is used to demonstrate how estimates of population benefit need to take into account patient eligibility, consent and adherence, and also how non-clinical data can inform the planning and development of health service interventions. PMID- 16255714 TI - Integrated support for Aboriginal tertiary students in health-related courses: the Pika Wiya Learning Centre. AB - The barriers to Indigenous people entering tertiary education, succeeding, and gaining employment in the health professions are broad and systemic. While efforts have been made to address these barriers, the number of Indigenous health professionals remains extremely low across Australia. The Pika Wiya Learning Centre in South Australia provides a range of practical, social, cultural, and emotional supports for tertiary students to increase the number of Indigenous health professionals, especially registered nurses, in the region. This paper reports on the Centre's strengths that may represent best practice in student support, and the obstacles to further development. PMID- 16255716 TI - Interactions between the plasma membrane and the antimicrobial peptide HP (2-20) and its analogues derived from Helicobacter pylori. AB - HP (2-20), a 19-residue peptide derived from the N-terminus of Helicobacter pylori ribosomal protein L1, has antimicrobial activity but is not cytotoxic to human erythrocytes. We synthesized several peptide analogues to investigate the effects of substitutions on structure and antimicrobial activity. Replacement of Gln16 and Asp18 with tryptophan [anal-3 (analogue-3)] caused a dramatic increase in lytic activities against bacteria and fungi. By contrast, a decrease in amphiphilicity caused by replacement of Phe5 or Leu11 with serine was accompanied by a reduction in antimicrobial activity. Analysis of the tertiary structures of the peptides in SDS micelles by NMR spectroscopy revealed that they have a well defined a-helical structure. Among the analogues, anal-3 has the longest a-helix, from Val4 to Trp18. The enhanced hydrophobicity and increased a-helicity results in enhanced antimicrobial activity in anal-3 without an increase in haemolytic activity. Fluorescence experiments proved that the bacterial-cell selectivity of the anal-3 peptide is due to its high binding affinity for negatively charged phospholipids in bacterial cells. Results showing the effect of spin-labels on the NMR spectra indicated that the side chains in the hydrophobic phase of the amphiphilic a-helix are buried on the surface of the micelle and the tryptophan indole ring is anchored in the membrane surface. Because anal-3 shows high selectivity towards bacterial and fungal cells, it may provide an avenue for the development of new antibiotics. PMID- 16255715 TI - A GPI-anchored alkaline phosphatase is a functional midgut receptor of Cry11Aa toxin in Aedes aegypti larvae. AB - A 65 kDa GPI (glycosylphosphatidyl-inositol)-anchored ALP (alkaline phosphatase) was characterized as a functional receptor of the Bacillus thuringiensis subsp. israelensis Cry11Aa toxin in Aedes aegypti midgut cells. Two (a 100 kDa and a 65 kDa) GPI-anchored proteins that bound Cry11Aa toxin were preferentially extracted after treatment of BBMV (brush boder membrane vesicles) from Ae. aegypti midgut epithelia with phospholipase C. The 65 kDa protein was further purified by toxin affinity chromatography. The 65 kDa protein showed ALP activity. The peptide displaying phages (P1.BBMV and P8.BBMV) that bound to the 65 kDa GPI-ALP (GPI anchored ALP) and competed with the Cry11Aa toxin to bind to BBMV were isolated by selecting BBMV-binding peptide-phages by biopanning. GPI-ALP was shown to be preferentially distributed in Ae. aegypti in the posterior part of the midgut and in the caeca, by using P1.BBMV binding to fixed midgut tissue sections to determine the location of GPI-ALP. Cry11Aa binds to the same regions of the midgut and competed with P1.BBMV and P8.BBMV to bind to BBMV. The importance of this interaction was demonstrated by the in vivo attenuation of Cry11Aa toxicity in the presence of these phages. Our results shows that GPI-ALP is an important receptor molecule involved in Cry11Aa interaction with midgut cells and toxicity to Ae. aegypti larvae. PMID- 16255717 TI - Intestinal alkaline sphingomyelinase hydrolyses and inactivates platelet activating factor by a phospholipase C activity. AB - Alkaline sphingomyelinase (alk-SMase) is a new member of the NPP (nucleotide pyrophosphatase/phosphodiesterase) family that hydrolyses SM (sphingomyelin) to generate ceramide in the intestinal tract. The enzyme may protect the intestinal mucosa from inflammation and tumorigenesis. PAF (platelet-activating factor) is a pro-inflammatory phospholipid involved in pathogenesis of inflammatory bowel diseases. We examined whether alk-SMase can hydrolyse and inactivate PAF. [3H]Octadecyl-labelled PAF was incubated with purified rat intestinal alk-SMase or recombinant human alk-SMase expressed in COS-7 cells. The hydrolytic products were assayed with TLC and MS. We found that alkSMase cleaved the phosphocholine head group from PAF and generated 1-O-alkyl-2-acetyl-sn-glycerol. Differing from the activity against SM, the activity against PAF was optimal at pH 7.5, inhibited by EDTA and stimulated by 0.1-0.25 mM Zn2+. The activity was abolished by site mutation of the predicted metal-binding sites that are conserved in all NPP members. Similar to the activity against SM, the activity against PAF was dependent on bile salt, particularly taurocholate and taurochenodeoxycholate. The V(max) for PAF hydrolysis was 374 mumol x h(-1) x (mg of protein)(-1). The hydrolysis of PAF and SM could be inhibited by the presence of SM and PAF respectively, the inhibition of PAF hydrolysis by SM being stronger. The PAF induced MAPK (mitogen-activated protein kinase) activation and IL-8 (interleukin 8) release in HT-29 cells, and chemotaxis in leucocytes were abolished by alk SMase treatment. In conclusion, alk-SMase hydrolyses and inactivates PAF by a phospholipase C activity. The finding reveals a novel function, by which alk SMase may counteract the development of intestinal inflammation and colon cancer. PMID- 16255718 TI - Existence of the school health coordinator in a frontier state. AB - The purpose of this study was to determine whether school districts in a rural western state employed school health coordinators or at least employed individuals who possessed the skills suggested for school health coordinators. Baseline data were collected soliciting the involvement of all 48 state school districts and the state girl's school (N = 49). Thirty-seven districts responded (75.5%). Identified school health coordinators were predominantly health and physical education teachers. Most coordinators spent minimal time (less than 10% of time) each week on school health coordination activities; nearly one half identified little or no coordination of school health efforts in their school districts; few identified personal involvement in budgetary matters concerning school health; and most identified their primary teaching responsibilities as the areas where the majority of their time was spent. Data collected help create a profile of the "typical" district level-school health coordinator in this frontier state and can assist in the development of future efforts aimed at school health coordination through the work of district-level school health coordinators. PMID- 16255719 TI - Substance abuse counseling services in secondary schools: a national study of schools and students, 1999-2003. AB - This study focuses on (a) American 8th-, 10th-, and 12th-grade students' access to, and use of, substance abuse counseling services via schools and (b) associations between such access and student substance use prevalence. From 1999 through 2003, student data were obtained from the Monitoring the Future study; and school data were obtained through the Youth, Education, and Society study, resulting in nationally representative data from 113,008 students in 855 public and private schools. Results indicate that in contrast to relatively stable student heavy-drug use prevalence rates, internal counseling availability and participation decreased significantly over time, as did reported student referral to external counseling. Availability of internal counseling, as well as student participation in both internal and external referrals, differed significantly by school characteristics--school level, grade size, sector, population density, school socioeconomic status, majority student body race/ethnicity, and geographical region. Student use of counseling services did not show any relationships with school-level heavy drinking rates; however, student participation in external counseling referrals was positively associated with school-level prevalence rates for the use of illicit drugs other than marijuana and showed indications of a similar relationship with marijuana prevalence rates. The decreasing access to, and use of, counseling, the lower probability of counseling availability in middle schools, and the lack of association between heavy-alcohol use rates and counseling services, all suggest missed opportunities and a greater need for counseling services to reduce high-risk drug use. PMID- 16255720 TI - Do social and behavioral characteristics targeted by preventive interventions predict standardized test scores and grades? AB - This study assessed whether characteristics of individuals that are predictors of youth problem behavior such as substance use, delinquency, and violence also predict academic achievement. Longitudinal data from 576 students participating in the Raising Healthy Children (RHC) project were analyzed. The RHC project is a study of students recruited from a suburban Pacific Northwest school district. Tenth-grade academic achievement was measured by scores on a standardized test administered to students in Washington State (as part of compliance with the No Child Left Behind Act) and by student self-report of grades. Measures of social and behavioral characteristics at seventh grade were based on data from student, parent, and teacher surveys. Researchers assessed overall correlations between 7th-grade predictors and 10th-grade academic achievement as well as partial correlations adjusted for demographic characteristics and scores on an earlier achievement test, the Comprehensive Test of Basic Skills, in 4th grade. Results indicated that higher levels of school bonding and better social, emotional, and decision-making skills were related to higher test scores and higher grades. Lower test scores and lower grades were predicted by elevated levels of attention problems, negative behavior of peers, and disruptive and aggressive behavior. Lower test scores also were predicted by early use of alcohol and cigarettes. These findings support the premise that school-based social development interventions that address specific risk factors, curb early manifestations of antisocial behavior, and promote school bonding and social and emotional skills are likely to improve student academic achievement. PMID- 16255721 TI - Risk behaviors for varying categories of disability in NELS:88. AB - A large body of research shows that youth with disabilities, who comprise about 13% of the country's school-aged population, report comparable to higher incidence rates of alcohol, tobacco, and other drug (ATOD) use than their peers. Furthermore, youth with disabilities who reported ATOD use or who engaged in binge drinking had significantly more negative educational outcomes and engaged in sexual activity at a younger age than nonusers. This study describes risk factors for substance use, personal characteristics, aspects of the attitudinal environment, and educational, employment, and social outcomes among youth across 6 categories of disability. Data came from the National Center for Education Statistics' National Education Longitudinal Study of 1988-2000 (NELS:88). The findings indicate that (a) youth with varying types of disabilities are relatively homogenous with respect to risk behaviors, personal characteristics, and outcomes; (b) youth with emotional, learning, or multiple disabilities may be at heightened risk for binge drinking and marijuana use; and (c) youth with emotional and multiple disabilities may be less likely to graduate from high school or its equivalent 8 years beyond the 12th grade. Based on these results and limitations of the NELS sampling strategy, appropriate interventions are discussed as well as the need for more definitive operational definitions for disabilities, specifically the biopsychosocial approach used by the International Classification of Functioning, Disability, and Health. PMID- 16255722 TI - The Carbohydrate Quandary: achieving health literacy through an interdisciplinary WebQuest. PMID- 16255723 TI - Genetics and the Immune Response. Abstracts of the 35th Annual Scientific Meeting of the Australasian Society for Immunology and the 14th International HLA and Immunogenetics Workshop, Melbourne, Australia, 4-8 Dec 2005. PMID- 16255724 TI - Respiratory function tests and their application. PMID- 16255725 TI - Abstracts of the 10th Congress of the Asian Pacific Society of Respirology (APSR), the 1st Congress of the APSR/ACCP. November 11-14, 2005. Guangzhou, China. PMID- 16255726 TI - Fordham, Jung and the self: a re-examination of Fordham's contribution to Jung's conceptualization of the self. AB - This paper is about Fordham's contribution to Jung's studies on the self. It opens with the epistemological dilemmas inherent in the subject, before moving on to an account of Fordham's research into the incompatible ways Jung used the term 'self'. There is a description of Fordham's model, which covers his concepts of the primary self, deintegration, reintegration, self objects, self representations, and individuation in infancy. There is a section which discusses areas in which Fordham apparently diverged from Jung, including how these were reconciled by Fordham's developmental approach. These areas include the definition of the self as totality or archetype, the mind-body relationship, the 'ultimate', the origins of the archetypes, and the primary self, the self and the sense of self. It concludes with an extension to Fordham's outline of a resolution to Jung's incompatible definitions. This draws upon the concept of the central archetype of order and how its unfolding is evidenced towards the end of the first year of infancy. PMID- 16255727 TI - Individuation: finding oneself in analysis--taking risks and making sacrifices. AB - This paper looks at some of the processes that are at work in finding oneself in analysis. It explores Jung's unique contribution to our thinking about the self and its dynamic of individuation. The author attempts to show how the Self, in its quest for consciousness, requires the surrendering of ego inflation--the narcissistic delusion that the ego is the self. A case is made for seeing analysis as an individuation process which offers the opportunity for experiences of a more authentic sense of oneself. Jung stated that individuation requires the ego to enter into service of the Self. For this to happen, the author argues that both patient and analyst must be prepared to make sacrifices and take risks. Using clinical examples, he illustrates that, although purposive, the Self can be experienced as violent and destructive if the ego is unable to facilitate its expression. This may result in an individuation crisis for both analyst and patient. The paper demonstrates how impasse in analysis can evoke the transcendent function, which also requires sacrifices to be made and risks to be taken for analysis to proceed. PMID- 16255728 TI - Sex, shame and the transcendent function: the function of fantasy in self development. AB - This paper explores a developmental approach to the sense of self-agency and to its influence on conscious and unconscious fantasy. I suggest that the emerging sense of self-agency offers an over-arching framework for our understanding of the nature and function of fantasy. In this context, intrusive and compulsive sexual fantasies which a person experiences as perverted and shameful, can be seen to serve differing psychic purposes, depending on the level of self-agency which is predominant. The fantasies can serve both as warning signals of the dangers of relationship and as opportunities for the mind to reflect on its own processes. Differing psychodynamic theories of fantasy are examined in terms of the developmental sense of self-agency that they represent. PMID- 16255730 TI - The unfolding and healing of analytic boundary violations: personal, clinical and cultural considerations. AB - Jungian analysts are not exempt from an unconscious engagement in a group complex. The author hypothesizes that there is a silent, dark legacy of belief in the superiority of men's judgment and the inferiority of women's, left by Jung, that has had a wounding impact on some Jungian analysands. Conscious and public mourning may be needed to heal our cultural complex. The author, a woman, traces the origins of her own patriarchal complexes and reveals how in her first analysis these mingled with the patriarchal complex shared by Jungian institute, her two male analysts and their former analyst, a pillar of the institute's community. Her first analyst aborted her analysis to begin a personal partnership with her. Her second analyst unconsciously colluded with the first analyst in not exploring this outcome as a violation. This resulted in a second compromised treatment. The senior analyst who had been these two analysts' own analyst was consulted, and he too failed to address the transgression. After experiencing severe symptomatology, the patient entered a third analysis with a woman where transference and regression were the focus. Eventually, meaning was found in the confrontations with the particular Jungian organization and its ethics committee, who acknowledged the first analyst's behaviour to be unethical. The author sees this process as a paradigm for the enactment and healing of a group complex. PMID- 16255729 TI - Sexual metaphor and the language of unconscious phantasy. AB - This paper investigates the attempt to find a 'bedrock' for psychic life in the idea of unconscious phantasy. Through a detailed examination of the development of the concept of unconscious phantasy, especially in Kleinian discourse, it is argued that unconscious phantasies are inherently metaphorical and have no 'concrete' existence in the unconscious. The use of unconscious phantasy as metaphor enables a 'two-way' form of interpretation that describes sexual behaviour and fantasy in terms of object relations (interpreting 'away from sex', while simultaneously describing object relations in terms of the sexual body (interpreting 'towards sex'). PMID- 16255731 TI - When does a dream begin to 'have meaning'? Linguistic constraints and significant moments in the construction of the meaning of a dream. AB - The paper investigates the origin of the meaning of dreams starting with a discussion of psychotic dreams. The author distinguishes the dream as dream from the remembered and the narrated. A two-step dream phenomenology is proposed that both acknowledges the objectivity of the dream and traces the origin of its meaning to its translation into language. Following a review of recent neurobiological theories of dreaming, the paper focuses on certain aspects of the psychoanalytical understanding of the dream phenomenon and in particular on the interpretation of dreams from the perspective of intersubjectivity and phantasy. PMID- 16255734 TI - How effective a recruiter to nursing are you? PMID- 16255735 TI - Bringing the professional challenges for nursing in Jordan to light. AB - The definition of a profession consists of contributing characteristics. Nursing must achieve these characteristics in order to confirm its professional status. These characteristics include: a standard of education; professional organizations; commitment; autonomy; continuing education; body of knowledge and competencies; social value; and a code of ethics. Nursing in Jordan faces a unique set of challenges to meet these criteria. The purpose of this discussion is to bring the professional challenges of nursing in Jordan to light, to motivate collaborative efforts to remediate them and to discuss the value of evidence-based practice in this process. The key to confirming Jordanian nurses' professional status is for nurses to seek every opportunity to prove themselves, earn respect, take control over their work, support their colleagues and to make informed decisions. In addition, a more active and effective role for the nursing organizations is necessary to change the public's view of nursing. Efforts to reform regulation also should be intensified. PMID- 16255736 TI - Nurses' moral agency in negotiating parental participation in care. AB - The purpose of this study was to investigate the issues for nurses in facilitating parental participation in the care of the hospitalized child. A qualitative study informed by grounded theory was undertaken. Nine nurses were recruited from an acute, high-dependency, 23-bed paediatric cardiac/renal unit in Melbourne, Australia. Data collection involved individual semistructured interviews, hospital policies related to family-centred care and a focus group interview. Constant comparative analysis was undertaken to develop an understanding of the data collected in the context of the nurses' experiences and the environment in which they work. Moral agency was identified as the central phenomenon of the study. Causal conditions related to this included the child's best interests, disputes about care and nurses' expectations. These causal conditions were seen to potentially lead to moral distress for the nurses. The coping mechanisms and strategies that affected moral agency have been identified. PMID- 16255737 TI - Improving transfer from the intensive care unit: the development, implementation and evaluation of a brochure based on Knowles' Adult Learning Theory. AB - This paper describes the development, implementation and evaluation of a transfer brochure for family members of patients in an intensive care unit (ICU) to improve patient transfer to a general ward. When family members fail to understand information, they respond in ways that affect patient recovery. The brochure was designed within Knowles' Adult Learning Theory framework and developed using a multidisciplinary team. A mixed design was used to collect data from families and nurses. Results indicate that the brochure helped nurses to address the individual family's issues during transfer from ICU. Furthermore, 95% of nurses (n = 33) recommended its introduction for all future transfers. Family members (n = 82) who received the brochure as part of their transfer were significantly more satisfied with all aspects of transfer than those who experienced ad hoc transfer methods (n = 80). These results provide strong support for Knowles' Adult Learning Theory as an educational foundation for adult learning. PMID- 16255738 TI - Jordanian women: perceptions and practices of first-time pregnancy. AB - A woman's child-bearing encounter is an experience that reflects the cultural beliefs and practices of the society. The purpose of this study was to describe the perceptions and practices of urban, low socioeconomic, Jordanian women (aged 18-30 years) in relation to their pregnancy career. An opportunistic sample consisting of 67 Muslim first-time pregnant women who followed up antenatal care at two maternity-care centres in East Amman was selected. A qualitative approach, including ethnographic semistructured interview, participant observation data from the households and clinics and oblique interviewing in random conversations initiated by the primigravidae, was conducted over 18 months. Women gave elaborative accounts of their pregnancy and childbirth expectations in narrative, phenomenological forms. Qualitative data analysis was performed concurrently with data collection, revealing the essential themes of immediacy of pregnancy; familial support and changing networks, especially for the after-birth period; fear of pain and medical interventions, all emphasizing the traditional and religious perceptions and practices in a familial context. PMID- 16255739 TI - Mutual suffering: a nurse's story of caring for the living as they are dying. AB - The aim of this study was to uncover the meaning of the lived experience of mutual suffering in relation to the care of a dying patient. The study took place within an acute medical ward in a district general hospital on the south coast of England as part of a reflective practice development programme. Parse's human becoming theory provided a framework for the study and Parse's research methodology was adopted. Understanding the nature of human relationships within nursing practice is central to nursing work, enabling patients and their health professional carers to live and work healthily in the context of human becoming. Illuminating mutual suffering through reflection enables nursing and health-care professionals to acknowledge the paradoxes of practice and, thus, create new strategies for the provision of care and the improvement of practice, so that quality of life is maximized for the patient and for themselves. PMID- 16255740 TI - 'They first killed his heart (then) he took his own life'. Part 1: a review of the context and literature on mental health issues for refugees and asylum seekers. AB - This is the first in a two-part series of papers examining mental health issues for refugees and asylum seekers. Beginning with the suicide of an asylum seeker in Scotland, the paper emphasizes mental health issues for adult and child asylum seekers, stress and memory, suicide, self-harm, risk and protective factors, compulsory health treatments and the prevention of mental illness. It sets the scene for the second paper by drawing implications for nursing practice in the community. Although most literature on refugee and asylum seeker mental health exists outside of nursing scholarship, a majority of the issues reviewed in this paper are mutual challenges for all in the health and helping professions. Nurses interested in refugee and migration issues face two intertwined challenges: that of how to assist migrants with their diverse mental health needs and how, at the same time, to contribute to a society that can promote mental health for all by taking on both the difficulties and opportunities posed by cultural diversity. PMID- 16255742 TI - Potential for recombinant Babesia bovis antigens to protect against a highly virulent isolate. AB - Two antigens from Babesia bovis,12D3 and 11C5, were expressed and purified as recombinant proteins in Escherichia coli and used to vaccinate groups of six Babesia-susceptible cattle. These were subsequently challenged with a highly virulent strain of B. bovis. All cattle showed symptoms of disease and most required treatment. Cattle vaccination groups receiving either 12D3 or 11C5 or a combination of both, reduced parasitaemia by approximately fourfold and a number of individual animals appeared to control the parasite infection. Control of parasites correlated with high monocyte numbers late in infection. The results thus confirm the potential usefulness of both antigens but also demonstrate the limitations of current formulations. PMID- 16255741 TI - Nonatopic allergen-independent mast cell activation in parasitized eosinophilic athymic rats. AB - It is possible to induce eosinophilia in congenitally athymic rats by infection with the parasite Ascaris suum. Athymic bronchial eosinophilia is associated with increased expression of IL-5 and eotaxin mRNA, and with the presence of residual T cells and mast cells. Anamnestic mastocytosis is particularly pronounced and in this study we examine the relationship between mast cell degranulation and IgE production in athymic rats following infection. Incubation of peritoneal mast cells from athymic rats with anti-IgE induced dose-dependent degranulation, as measured by histamine release. However, the failure of mast cells from infected athymic rats to degranulate following incubation with all but one of the parasite antigens selected confirms the absence of a specific IgE response. In contrast, all agonists induced degranulation in euthymic rats. The only parasite-derived factor to induce histamine release in all mast cells was Ascaris body fluid, which contains an element capable of inducing IgE-independent degranulation. Furthermore, increased levels of rat mast cell protease II in athymic bronchoalveolar lavage fluid suggest degranulation of mast cells by IgE and allergen independent means in vivo. We believe that the development of eosinophilic lung inflammation in the absence of specific IgE makes this a prototype for investigating the immunological mechanisms underlying nonatopic asthma. PMID- 16255743 TI - Ribavirin up-regulates IL-12 p40 gene expression and restores IL-12 levels in Leishmania-treated PBMCs. AB - Ribavirin, a nucleoside analogue that interferes with viral mRNA synthesis and inhibits the replication of RNA and DNA viruses, has been recently proposed as an effective immune response modulator. In the present report, we studied the effect of ribavirin on IL-12 p40 gene expression in peripheral blood mononuclear cells (PBMCs) of healthy subjects. We also studied ribavirin effects on PBMCs activated with lipopolysaccharide (LPS) and phytohaemagglutinin (PHA) and treated with Leishmania donovani antigens. We provide evidence that ribavirin was able to up regulate IL-12 p40 gene expression and to restore levels of IL-12 p40 gene expression and IL-12 secretion in fully activated PBMCs that were strongly inhibited by L. donovani antigens. Because effective management of leishmanial disease is usually associated with a prevalent T-helper 1 immune response with elevated production of IL-12,our preliminary results may be of particular interest, provided that they will be confirmed by further in vitro and in vivo studies, when considering a possible use of ribavirin as adjuvant in severe leishmanial disease. PMID- 16255745 TI - Kinetics of immunoglobulin G, M, A and IgG subclass responses in experimental intravaginal trichomoniasis: prominence of IgG1 response. AB - Trichomoniasis, the most common nonviral sexually transmitted disease, is caused by infection with the protist Trichomonas vaginalis. The clinical spectrum varies from an asymptomatic state to mild, moderate or severe symptoms. However, the exact factors leading to varied symptomatology have not been well elucidated. The mouse is a useful experimental model for intravaginal trichomoniasis, for understanding the role of local immune responses in the pathogenesis and varied symptomatology of this disease. The present study reports anti-Trichomonas IgG, IgM, IgA and IgG subclass antibody responses on different post-infection days (3rd, 7th, 14th, 21st and 28th) in serum and vaginal washes of mice infected intravaginally with T. vaginalis isolates from 15 symptomatic and 15 asymptomatic women. Successful intravaginal infection was established by inoculating T. vaginalis in BALB/c mice pre-inoculated with Lactobacillus acidophilus and pretreated with oestradiol. A significant increase in parasite load was observed on the 14th post-infection day (p.i.d.) in mice inoculated with T. vaginalis isolates from symptomatic women as compared to asymptomatic women (P < 0.001), followed by reduction until the 28th p.i.d. (P < 0.05). A significant increase in specific IgG (P < 0.001) and in particular IgG1 (P < 0.001) and IgM (P < 0.01) responses was observed on the 14th p.i.d. in serum and vaginal washes of mice infected with T. vaginalis isolates from symptomatic women as compared to asymptomatic women. Significant increases in specific IgG, IgM and IgG1 responses was observed on the 14th p.i.d. in serum samples as compared with vaginal washes of mice infected with T. vaginalis isolates from symptomatic and asymptomatic women (P < 0.01), whereas no significant difference was observed in IgA, IgG2a, IgG2b and IgG3 antibody responses. The study indicates that specific IgG, particularly IgG1 and IgM, may be playing a role in establishing symptomatic infection. PMID- 16255744 TI - Cytopathic changes and pro-inflammatory cytokines induced by Naegleria fowleri trophozoites in rat microglial cells and protective effects of an anti-Nfa1 antibody. AB - Naegleria fowleri, a free-living amoeba, causes fatal primary amoebic meningoencephalitis in experimental animals and humans. The nfa1 gene (360 bp) was previously cloned from a cDNA library of pathogenic N. fowleri by immunoscreening, and produced a 13.1-kDa recombinant protein that showed pseudopodia-specific localization by immunocytochemistry. On the basis of an idea that the pseudopodia-specific Nfa1 protein seems to be involved in the pathogenicity of N. fowleri, the cytopathic activity of N. fowleri trophozoites co-cultured with rat microglial cells was observed, and the effects of an anti Nfa1 antibody in a co-culture system were elucidated. Using light, scanning and transmission electron microscopy, it was seen that N. fowleri trophozoites in contact with microglial cells produced vigorous pseudopodia and a food-cup structure. Microglial cells were destroyed by N. fowleri trophozoites as seen from necrotic cell death in a time-dependent manner. In a(51)Cr release assay, N. fowleri showed 17.8%, 24.9%, 54.6% and 98% cytotoxicity against microglial cells at 3, 6, 12 and 24 h post-incubation, respectively. However, when anti-Nfa1 antibody was added in a coculture system, N. fowleri cytotoxicity was reduced to 15.5%, 20.3%, 46.7% and 66.9%, respectively. Moreover, microglial cells co cultured with N. fowleri trophozoites secreted the pro-inflammatory cytokines, TNF-alpha, IL-1beta and IL-6. In the presence of anti-Nfa1 antibody, the secretion of TNF-alpha was slightly, but not significantly, decreased. PMID- 16255746 TI - Modulation of murine cellular immune response and cytokine production by salivary gland lysate of three sand fly species. AB - Saliva of sand flies (Diptera: Phlebotominae) plays an important role in transmission of Leishmania parasites by modulating host immune response. However, because of the different protein compositions of saliva, the immunomodulatory effects may vary among sand fly species. We have therefore analysed and compared the immunomodulation effects of salivary gland lysate (SGL) of three different sand flies. Spleen cells from BALB/c mice were incubated with SGL of Phlebotomus papatasi, P. sergenti or Lutzomyia longipalpis. Concanavalin A-stimulated lymphocyte proliferation was significantly suppressed with SGLs of all three sand fly species and all SGL doses tested. This result indicates that saliva from different sand fly species is able to suppress host proliferative response even to the potent mitogen. In parallel experiments, we analysed the effect of SGL on IFN-gamma, IL-2, and IL-4 production; in mitogen-stimulated cells SGLs markedly inhibited IFN-gamma production in all intervals tested (reduced up to 31%) and to a lesser degree impaired production of the other two cytokines as well. Despite some species-specific differences in the intensity of immunomodulatory effects, saliva of all sand fly species modulated cell proliferation as well as cytokine production in a similar way. PMID- 16255747 TI - Noninvasive electrocardiology in 2005 and beyond: letter from ISHNE President. PMID- 16255748 TI - Noninvasive risk stratification of subjects with a Brugada-type electrocardiogram and no history of cardiac arrest. AB - BACKGROUND: Recent studies suggest that the Brugada-type electrocardiogram (ECG) is much more prevalent than the manifest Brugada syndrome. Although invasive electrophysiologic investigations have been proposed as a risk stratifier, their value is controversial, and alternative noninvasive techniques may be preferred. We sought a noninvasive strategy to detect a high-risk group in a long-term follow-up study of subjects with a Brugada-type ECG, and no history of cardiac arrest. METHODS: This study enrolled 124 consecutive subjects with a Brugada-type ECG. Prognostic indices included: age, sex, a family history of sudden death, syncopal episodes, a spontaneous coved-type ST-segment elevation, maximal magnitude of ST-segment elevation, a spontaneous change in ST segment, a mean QRS duration, maximal QT interval, QT dispersion, late potentials (LP) by signal averaged ECG, and microvolt T-wave alternans. RESULTS: Of the 124 subjects, 20 consenting subjects had an implantable defibrillator before follow-up. During a 40 +/- 19-month follow-up, 12 subjects (9.7%) reached one of the endpoints (sudden death or ventricular tachyarrhythmia). Of the 12 risk indices, a family history of sudden death, syncopal episodes, a spontaneous coved-type ST-segment elevation, a spontaneous change in ST segment, and LP had significant values. In multivariate analysis, a spontaneous change in ST segment had the most significance (a relative hazard, 9.2; P = 0.036). Combined assessment of this index and other significant indices obtained higher positive predictive values (43-71%). CONCLUSIONS: A spontaneous change in ST segment is associated with the highest risk for subsequent events in subjects with a Brugada-type ECG. The presence of syncopal episodes, a history of familial sudden death, and/or LP may increase its value. PMID- 16255749 TI - Safe scanning, but frequent artifacts mimicking bradycardia and tachycardia during magnetic resonance imaging (MRI) in patients with an implantable loop recorder (ILR). AB - BACKGROUND: Patients with implantable devices are generally not permitted to undergo magnetic resonance imaging (MRI) because of potentially deleterious interactions. Little has been reported regarding the safety and effects of MRI scanning of patients with implantable loop recorders (ILRs). We evaluated the safety of scanning patients with ILRs and the output of the ILR after undergoing MRI. METHODS: Ten patients underwent 11 MRI scanning events. All patients had Reveal Plus (Medtronic, Minneapolis, MN) ILRs. Seven cranial, two lumbar-spine, one shoulder, and one knee MRI were performed. All of the MRIs were performed with the understanding that the patient had an ILR. In each patient, the ILR was cleared moments before the scan and the integrity of the signal and time date stamp were verified. The devices were reinterrogated immediately after MRI in 10 patients and two days post MR scanning in one patient. Each patient was questioned post MRI regarding any symptoms experienced during the scan. RESULTS: Both tachy and bradyarrhythmias appeared as artifacts as a result of ILR exposure to MRI. Post MRI, none of the ILRs showed diminished signal integrity, altered programmed parameters, diminished battery status, inability to communicate or be reprogrammed. No sensations of tugging or warmth at the implant site were noted. CONCLUSION: MRI was performed in ILR patients without harm to the patient or permanent damage to the ILR. MRI scanning of the Reveal appears safe. Artifact mimicking an arrhythmia was common, however, and must be excluded in any ILR patient undergoing MRI to avoid mistakenly attributing a syncopal episode, or palpitations to the artifacts produced from MRI exposure. PMID- 16255750 TI - Effects of mobile telephones on the function of implantable cardioverter defibrillators. AB - OBJECTIVE: We investigated whether mobile telephones affect the function of implantable cardioverter defibrillators (ICDs). BACKGROUND: It is well known that electromagnetic fields can affect medical devices. METHODS: The study included 43 patients with ventricular tachycardia and/or fibrillation treated with transvenous pectoral ICDs. Testing was done under continuous electrocardiograph monitoring under supervision of an ICD programmer. Initially, each patient was tested during spontaneous rhythm. Then the ICD was programmed to a pace rhythm higher than the patient's heart rate, and the tests were repeated at paced rhythm. In 7 patients, tests were performed during the implantation procedure as well. In 3 of the patients, only a single defibrillation zone was active. The other 40 patients had one or more active ventricular tachycardia zones. Two mobile phones (both GSM 900 MHz) were positioned 50 cm away from the implanted device in opposite directions and switched on. Communication was established between these phones, two investigators had a 20-second conversation, and then the phones were switched off. The same procedure was repeated at 30, 20, and 10 cm away from the implantation site, respectively. Finally, the procedure was performed with the antennae of both phones touching the device pocket. In the above-mentioned 7 cases where testing was done during implantation of the ICD, a call was made from one phone to the other, ringing occurred for 5 seconds, and then two investigators conversed while the device was implanted. RESULTS: There was no change in the function of the ICDs during any of the phone testing procedures. In 5 cases, artifacts were noted on the surface electrocardiographic (ECG) screen of the programmer during the tests, but no such changes were observed on the simultaneous intracardiac ECGs. CONCLUSION: The results of the study suggest that mobile phones have no effects on ICD function. PMID- 16255751 TI - Prediction of the recurrence of atrial fibrillation after successful cardioversion with P wave signal-averaged ECG. AB - BACKGROUND: The recurrence of atrial fibrillation (AF) was often observed after cardioversion. METHODS: In our study, a P wave triggered P wave signal-averaged ECG (P-SAECG) was performed on 118 consecutive patients 1 day after successful electrical cardioversion in order to evaluate the utility of this method to predict AF after cardioversion. We measured the filtered P wave duration (FPD) and the root mean square voltage of the last 20 ms of the P wave (RMS 20). RESULTS: During a 1-year follow-up, a recurrence was observed in 57 patients (48%). Patients with recurrence of AF had a larger left atrial size (41.9 +/- 4.0 vs 39.3 +/- 3.1 mm, P < 0.0003), a longer FPD (139.6 +/- 16.0 vs 118.2 +/- 14.1 ms, P < 0.0001), and a lower RMS 20 (2.57 +/- 0.77 vs 3.90 +/- 0.99 microV, P < 0.0001). A cutoff point (COP) of FPD > or =126 ms and RMS 20 < or =3.1 microV could predict AF with a specificity of 77%, a sensitivity of 72%, a positive value of 75%, a negative predictive value of 75%, and an accuracy of 75%. A stepwise logistic regression analysis of variables identified COP (odds ratio 9.97; 95% CI, 4.10-24.24, P < 0.0001) as an independent predictor for recurrence. CONCLUSIONS: We conclude that the probability of recurrence of AF after cardioversion could be predicted by P-SAECG. This method seems to be appropriate to demonstrate a delayed atrial conduction that might be a possible risk factor of reinitiation of AF. PMID- 16255752 TI - A Latin American registry of implantable cardioverter defibrillators: the ICD LABOR study. AB - OBJECTIVE: Despite the progress that has been reached in emergency medical systems and resuscitation, sudden cardiac death (SCD) continues to be the major cause of the death, and remains a significant public health problem. In this publication we are reporting our Latin American experience in the secondary prevention of SCD, by means of an ongoing registry involving seven Latin American countries and 770 patients. METHODS: Every individual within the present registry to date has presented with antecedents of aborted sudden death or cardiac arrest due to ventricular tachycardia or ventricular fibrillation. Patients included have fulfilled the Class I indication for implantable cardioverter defibrillator (ICD) and they were implanted with a Biotronik ICD (all models). The study was not sponsored by Biotronik, nor did they have access to the data. A specific protocol was designed for implantation and follow-up of patients. The database was completely registered through the Internet and a personal password was assigned to each group of investigators. The primary end point was death from all causes. Secondary end points were SCD and death due to congestive heart failure (CHF). RESULTS: The etiology of cardiac disease was found to be predominantly coronary artery disease (CAD) 39.7% (306 patients), followed by Chagas disease (ChD), 26.1% (201 patients), and idiopathic dilated cardiomyopathy (DCM), 17% (131 patients). Any remaining pathologies were included as miscellaneous 13.2% (101 patients). In 31 patients (4%) the etiology was unknown. The age did not differ within the principal pathologies, but was significantly older than the miscellaneous group (62.0 +/- 11.3 years vs 48.2 +/- 18.9 years, P < 0.0001). The follow-up period was 27 +/- 25 months (1-113 months) for the whole group. The mortality in functional classes I-II was significantly lower than mortality for functional classes III-IV (relative risk 1.46, CI 95%, P < 0.0001). Mean left ventricular ejection fraction (LVEF) for the whole group was 37.7 +/- 14.3%. Male LVEF was 36.1 +/- 14.1% and female LVEF was 42.2 +/- 13.8% P < 0.0001. During the follow-up period, 130 deaths were reported (global mortality 16.9 +/- 9.7%), out of which 84 (64.6%) were attributed to cardiac causes (10.9 +/- 5.1% of the total population). The annual adjusted cardiac mortality was 5.2 +/- 1.72% (range 3.5 7.0%). Among cardiac deaths the most common cause was progressive heart failure, 48 patients (57%) including 3 patients with pulmonary embolism. The second main cause of cardiac death was SCD, 36 patients (43%), including 4 patients with electrical storm and 3 patients with electromechanical dissociation after multiple shock therapy treatments. CONCLUSIONS: Despite the differences in terms of pathologies between the ICD-LABOR (Latin American bioelectronic ongoing registry) and randomized ICD trials, a parallel evolution in all cause mortality and cardiac mortality was observed. Independent risk factors for mortality included age >70 years, male gender, NYHA III/IV, and ejection fraction <0.30. The etiology of heart disease (Chagas vs Coronary Disease) was not found to be a risk factor. PMID- 16255753 TI - Differences in outcomes between patients treated with single- versus dual-chamber implantable cardioverter defibrillators: a substudy of the Multicenter Automatic Defibrillator Implantation Trial II. AB - OBJECTIVES: We sought to evaluate the influence of single- versus dual-chamber implantable cardioverter defibrillators (ICDs) on the occurrence of heart failure and mortality as well as appropriate and inappropriate ICD therapy in the Multicenter Automatic Defibrillator Implantation Trial II (MADIT-II). BACKGROUND: In MADIT-II, ICD therapy in patients with a prior myocardial infarction and ejection fraction < or =0.30 was associated with a 31% reduction in risk of mortality when compared to conventionally treated patients. An unexpected finding was an increased occurrence of hospitalization for heart failure in the ICD group. METHODS: Data from 717 patients randomized to ICD therapy with single- or dual-chamber pacing devices in MADIT-II were retrospectively analyzed. Endpoints selected for analysis included death from any cause, new or worsening heart failure requiring hospitalization, death or heart failure, appropriate therapy for ventricular tachycardia (VT) or ventricular fibrillation (VF), and inappropriate ICD therapy for atrial fibrillation or supraventricular tachycardia. RESULTS: A total of 404 single-chamber ICDs (S-ICDs) and 313 dual chamber ICDs (D-ICDs) were implanted. Patients receiving D-ICDs were at a higher risk at baseline than those receiving S-ICDs, with older age, higher NYHA class, more frequent prior CABG, wider QRS complex, more LBBB, higher BUN level, a history of more atrial arrhythmias requiring treatment, and a longer time interval from their index myocardial infarction to enrollment. While there was a trend toward an increase in adverse outcomes in the D-ICD group, no statistically significant differences in heart failure or mortality were observed between S-ICD versus D-ICD groups. CONCLUSIONS: Patients with D-ICDs had a nonsignificant trend toward higher mortality and heart failure rates than patients with S-ICDs. PMID- 16255754 TI - Short QT syndrome. AB - Short QT syndrome (SQTS) is an inheritable primary electrical disease of the heart, discovered in 1999. It is characterized by an abnormally short QT interval (<300 ms) and a propensity to atrial fibrillation and sudden cardiac death (SCD). Like in the case of long QT syndrome there is more than one genetic mutation that can lead to a short QT interval in the ECG and so far two have been identified. Shortening of the effective refractory period combined with increased dispersion of repolarization is the likely substrate for reentry and life threatening tachyarrhythmias. Only 22 people have been classified as having SQTS: 15 from the actual measurement of a short QT interval in their ECG and 7 by history, all having died from SCD. It is very likely that several cases, especially among children, have been overlooked, since the shortness of the QT interval only becomes apparent at heart rates <80 beats/min. The best form of treatment is still not known, but prevention of atrial fibrillation has been accomplished by propafenone, and an implantable cardioverter defibrillator is recommended for prevention of SCD. PMID- 16255755 TI - Vagal afferent stimulation as a cardioprotective strategy? Introducing the concept. AB - The effect of vagal afferent signaling on cardioinhibition has been well known for over 130 years. Both experimental and clinical studies have demonstrated not only the potential adverse effect of unrestrained sympathoexcitation in high risk patients with ischemic heart disease but the potential for cardioprotection by programmed vagal activity. The vasodepressor and negative chronotropic effects of efferent vagal stimulation has been a cause for concern. However it is becoming clear that favorable shifts towards increased cardiac vagal modulation can be achieved by vagal afferent nerve stimulation. This phasic effect appears to operate though central medullary pathways. Thus by engaging vagal afferent fibers in humans there is the possibility that one can exploit the benefits of central cardioinhibition without adversely affecting heart rate, respiration or hemodynamics. This commentary explores the background and rationale for considering vagal afferent stimulation as a plausible cardioprotective strategy. PMID- 16255756 TI - Silymarin treatment of viral hepatitis: a systematic review. AB - Silymarin from the milk thistle herb (Silybum marianum) is used by many patients with chronic viral hepatitis, but its efficacy remains unknown. We performed a systematic review of silymarin for the treatment of chronic viral hepatitis B and C. An exhaustive search strategy identified 148 papers that studied silymarin compounds in liver disease. Of these, four trials included patients with hepatitis C, one included hepatitis B patients, and two, unspecified chronic viral hepatitis. However, only one trial exclusively studied patients with hepatitis C, and none involved patients with only hepatitis B. Silymarin treatment resulted in a decrease in serum transaminases compared with baseline in four studies, and compared with placebo in only one study. There is no evidence that silymarin affects viral load or improves liver histology in hepatitis B or C. No studies were found that investigated the use of silymarin concomitantly with interferon, nucleoside analogues, or other conventional treatments for hepatitis B or C. In conclusion, silymarin compounds likely decrease serum transaminases in patients with chronic viral hepatitis, but do not appear to affect viral load or liver histology. Nevertheless it may be worthwhile to determine its effects in conjunction with standard antiviral treatment. PMID- 16255757 TI - The -670A > G polymorphism in the promoter region of the FAS gene is associated with necrosis in periportal areas in patients with chronic hepatitis C. AB - Evidence suggests that apoptosis of liver cells may play a significant role in the pathogenesis of hepatitis C virus (HCV) infection. One of the best characterized apoptotic pathway is that mediated by the death receptor Fas. Fas expression has been found to be up-regulated on hepatocytes in chronic HCV infection, particularly in periportal areas. Recently, two polymorphisms have been identified in the promotor region of the FAS gene, -1377G > A and -670A > G. We have evaluated the involvement of these variants in the susceptibility to HCV infection, the severity of liver damage and progression of fibrosis in chronic hepatitis C. A cohort of 197 patients with chronic hepatitis C and 100 controls were analysed for both polymorphisms by Fluorescence Resonance Energy Transfer using specific probes and the Lightcycler system. In addition, liver biopsies were taken in 167 patients and scored using the Knodell classification system. We have found that the allele frequencies and the distribution of both polymorphisms do not differ significantly in the HCV cohort and in the control population. Thus, none of the polymorphisms seems to be related with susceptibility to HCV infection. However, we have examined the possible association between the two variants and the grade of necroinflammatory activity and the stage of fibrosis and we have detected an under-representation of the -670A > G variant among those patients with higher Knodell's scores (P = 0.049) and necroinflammatory activity (P = 0.036). The -670A allele was associated with higher levels of periportal necrosis (P = 0.012). In conclusion, our findings suggest an association between the -670A > G polymorphism and the grade of necrosis in periportal areas in patients with chronic hepatitis C. PMID- 16255758 TI - Viral evolution of hepatitis C in injection drug users. AB - Injection drug users represent the largest cohort of patients with established hepatitis C virus (HCV) infection as well as the group that is at highest risk for new infections. Most published studies have focused on the clinical consequences of established HCV infection and have not examined the consequences of new infection. The aim of the current study was to measure the virological consequences of HCV in patients with ongoing injection drug use that might pose a risk for new and/or for superinfection with additional strains of HCV. We examined the following groups: (a) those with resolved HCV infection with ongoing injection drug use, (b) those with chronic infection who continued to inject and (c) those with chronic infection who no longer injected. Our study demonstrated a spectrum of responses. The majority of patients appeared to be 'protected' from new infection. None of six patients with resolved infection had detectable HCV RNA by quantitative or qualitative PCR when followed for 1 year. Similarly, despite ongoing injection drug use, no patient with persistent infection had a 'switch' in HCV genotype indicative of possible superinfection. Virological analysis of HCV quasispecies to detect possible infection with new variants of HCV in patients with apparently 'stable' infection, indicated divergence of virus over time, divergence that was unrelated to injection drug behaviour. Thus, patients with ongoing or prior HCV infection appear to develop immunity that protects against further infection with HCV despite repeated exposure. PMID- 16255759 TI - Identification of glycosaminoglycan-binding sites within hepatitis C virus envelope glycoprotein E2*. AB - Heparan sulphate is one of the candidate receptors for hepatitis C virus (HCV). Envelope glycoproteins of HCV have been proposed to be responsible for recognition and binding with cell receptors. They are characterized by great genetic polymorphism. In this study the mapping of regions with glycosaminoglycan binding properties within HCV envelope proteins has been undertaken. We prepared a set of overlapping peptides corresponding to conserved regions of these envelope proteins and analysed them by solid phase heparin-binding assay. The search for established glycosaminoglycan-binding motifs in the HCV envelope proteins showed the absence of the sites corresponding to the glycosaminoglycan binding patterns in consensus sequence. We identified one highly conserved and two less conserved heparin-binding sequences within the envelope protein E2 based on solid phase assay results. We did not find any differences in binding efficiency of these peptides with heparin, heparan sulphate or dextran sulphate. Our data supported the specific association between HCV envelope protein E2 and cell surface glycosaminoglycans. We hypothesize that identified regions from E2 can contribute to HCV binding to cell surface glycosaminoglycans. PMID- 16255760 TI - Viral load change and sequential evolution of entire hepatitis C virus genome in Irish recipients of single source-contaminated anti-D immunoglobulin*. AB - In hepatitis C virus (HCV) infection, serum viral load is important in the prediction of therapeutic efficacy. However, factors that affect the viral load remain poorly understood. To identify viral genomic elements responsible for the viral load, we investigated samples from a population of Irish women who were iatrogenically infected from a single HCV source by administration of HCV 1b contaminated anti-D immune globulin between 1977 and 1978 (Kenny-Walsh, N Engl J Med 1999; 340: 1228). About 15 patients were divided into two groups, viral load increasing group (11 patients) and decreasing group (4 patients). Pairs of sera were collected from each patient at interval between 1.1 and 5.8 years. Full length sequences of HCV genome were determined, and analyzed for changes in each patient. Sliding window analysis showed that the decreasing group had significantly higher mutation rates in a short segment of NS5B region that may affect the activity of RNA-dependent RNA polymerase. By comparing each coding regions, significantly higher mutation numbers were accumulated in NS5A region in the increasing group than the decreasing group (0.92 vs 0.16 nucleotides/site/year, P = 0.021). The mutation in certain positions of the HCV genome may be determinant factors of the viral load in a relatively homogeneous patient population. PMID- 16255761 TI - Immune tolerance to hepatitis C virus acquired during engraftment of bone marrow transplant. AB - The CD4+ T-cell response appears to be important for clearance of hepatitis C virus (HCV) in the majority of individuals. We have recently described a series of human leucocyte antigen (HLA)-DR11-restricted T-cell epitopes derived from HCV proteins which enables distinct populations of memory CD4+ T cells to be detected and counted in all nonviraemic HCV subjects. We examined the case of an HLA-DR11+ recipient of a haematopoietic stem-cell transplant who was concurrently infected with HCV from an HLA-DR11+ donor sibling. An acute HCV hepatitis developed and was treated with type I interferon. After successful viral clearance, the recipient demonstrated a selective lack of HCV epitope-specific CD4+ T cells and absence of serological responses compared with the treated donor. The recipient had no evidence of any nonspecific immunosuppression. The subsequent effects of concurrent infection during immune reconstitution are not known in adult humans, but data from murine models suggest this can lead to a skewing of the T-cell repertoire because of thymic selection. From the above observations, it is plausible that the introduction of foreign viral antigen into the thymus may lead to subsequent acquired central tolerance. PMID- 16255762 TI - Geographic distribution, virologic and clinical characteristics of hepatitis B virus genotypes in China. AB - The significance of hepatitis B virus (HBV) genotypes for the heterogeneity of chronic HBV infection and severity of liver disease is not well understood. The aim of this study was to determine the distribution and virologic characteristics of HBV genotypes in China and possible association with the diversity of liver disease. The study includes 1096 chronic HBV carriers from nine provinces in China. We collected clinical and laboratory data and analysed the HBV strains in sera by polymerase chain reaction-restriction fragment length polymorphism (PCR RFLP) and nucleotide sequencing techniques. The most common HBV genotypes were B (41%) and C (53%), while genotypes A and D were also found. A North-South divide was identified in genotype B and C distribution - genotype C was predominant in northern China, while genotype B was more prevalent in southern provinces. Patients with genotype B were younger than those with genotype C, and had a lower prevalence of HBeAg - 65%vs 72%, respectively (P = 0.03). However, the severity of liver disease did not differ significantly between patients infected with genotype B or C - neither when comparing liver function tests (1024 patients), nor hepatic inflammation and fibrosis (264 patients). Amongst 47 patients with genotype D (by PCR-RFLP), 37 (79%) were infected with a new subtype (designated Dc), having a recombination fragment from genotype C precore/core region. This is the first large-scale HBV genotype study from China and convincing documentation of the North-to-South gradient of genotypes C vs B in this country. HBV DNA recombination over the surface and precore/core genes increases the diversity of HBV strains and may have diagnostic and clinical implications. PMID- 16255763 TI - Pathways of care and resource utilization in a national cohort of patients with transfusion-acquired hepatitis C. AB - The Hepatitis C Strategy and Action Plan for England recommend that all individuals testing positive for hepatitis C virus (HCV) should be referred to a specialist centre for assessment and care. One key aim is to reduce the number of people progressing to liver disease and therefore reduce the associated costs. The aims of this paper are to describe the care pathways and evaluate resource utilization in a cohort of 826 patients with transfusion-acquired hepatitis C enrolled in the HCV national register. We reviewed data extracted from patient notes to establish pathways of care since HCV-positive diagnosis through to May 2002, and to document all treatment, liver biopsy and hospital usage for each patient. Type of care was classified into specialist-interest in HCV-related care, other-hospital care or general practitioner (GP)-led care. Over 70% of patients were referred to specialist care following HCV diagnosis. Patients who were older or who had normal liver function were less likely to be referred to specialist-care. Between first diagnosis and May 2002, no patients were referred from GP to specialist-care. Less than half of this cohort had undergone liver biopsy and only 18% had been treated. Younger patients and those with abnormal liver function were more likely to have undergone liver biopsy and to have received treatment. Analysis of care histories of patients with transfusion acquired hepatitis C suggest that changes are needed in the care and management of patients with HCV infection, if the recommendations of the HCV strategy and action plan are to be fully implemented. PMID- 16255764 TI - Role of nucleic acid testing in cadaver organ donor screening: detection of hepatitis C virus RNA in seropositive and seronegative donors. AB - Hepatitis C virus (HCV) transmission by both seropositive and seronegative cadaver organ donors has been documented, yet nucleic acid testing is not routinely used to identify active infection in these donors prior to transplantation. Between November 2001 and February 2004, we screened 1445 cadaver organ donors for anti-HCV antibodies with either HCV EIA-2.0 (Abbott Diagnostics, Chicago, IL, USA) and/or Ortho HCV Version 3.0 ELISA (Ortho-Clinical Diagnostics, Raritan, NJ, USA) and confirmed seropositive samples with Chiron RIBA3.0 SIA (Chiron Corporation, Emeryville, CA, USA). Samples with sufficient volume (n = 726) were tested by the VERSANT HCV [transcription-mediated amplification (TMA)] Qualitative assay (Bayer Healthcare LLC, Tarrytown, NY, USA) which can be performed in approximately 5 h. Those with detectable HCV RNA and sufficient volume were quantified by the VERSANT HCV 3.0 (bDNA) Assay (Bayer Healthcare LLC) and/or the HCV RNA TMA Quantitative Assay (n = 23) and genotyped (n = 57). Seventy-seven of 1445 (5.3%) donors were seropositive, reactive by either one or both anti-HCV assays. Fifty-two of 63 (82.5%) of the seropositive samples had detectable HCV RNA and were genotyped. Seventeen of these samples had quantifications ranging from 128,123 to >7,692,307 IU/mL. Six of 663 (0.9%) seronegative samples had detectable HCV RNA. Their quantifications ranged from <9.3 to 1,464,799 IU/mL, and five of these six were successfully genotyped. As HCV RNA was demonstrated in samples from both our seropositive and seronegative cadaver organ donors, we are now incorporating nucleic acid testing into our donor screening/diagnostic algorithm. PMID- 16255765 TI - Hepatic expression of the proliferative marker Ki-67 and p53 protein in HBV or HCV cirrhosis in relation to dysplastic liver cell changes and hepatocellular carcinoma. AB - To evaluate hepatic expression of the nuclear proliferative marker Ki-67 and the p53 oncoprotein in hepatitis B virus (HBV)/HCV cirrhosis in relation to dysplastic liver cell changes and hepatocellular carcinoma (HCC). We studied needle liver biopsies from 107 patients with cirrhosis and no HCC (52 HBV, 55 HCV) who had been assessed for protocol studies, and 57 cirrhotic patients with HCC (40 HBV, 17 HCV). We evaluated small and large cell dysplastic changes along with the expression of Ki-67 and p53 by immunohistochemistry. The labelling index (LI) was defined as the proportion (%) of positive-stained nuclei of the 500 measured. Large and small cell dysplastic changes were observed in 12 and 9% of specimens respectively. Only small cell changes were associated with Ki-67 expression. Ki-67 LI was 5.50 +/- 5.7 in cirrhosis (13.90 +/- 3.84 in those with small cell dysplastic changes vs 4.64 +/- 4.98 in those without, P < 0.01), 10.2 +/- 5.95 in cirrhosis with HCC (P < 0.05) and 18.56 +/- 10 in HCC (P < 0.01). Neither the presence of small cell dysplastic changes nor the expression of Ki-67 was related to severity or aetiology of cirrhosis. Expression of p53 was observed in 30% of the non-tumorous and in 53% of the neoplastic tissue obtained from patients with HCC, with no differences between HCV and HBV. Ki-67 and p53 expression was associated with the tumour grade (P < 0.001). Our observations clearly demonstrate the association between the proliferation activity and the morphological changes in the cirrhotic liver from the non-dysplastic to dysplastic lesion to HCC. They also support the hypothesis that p53 alterations are a rather late event in carcinogenesis and related to HCC grade. And finally, they suggest that the final steps of hepatocarcinogenesis are common and independent of the aetiology of the chronic viral infection. PMID- 16255766 TI - Hepatitis B surface antigenaemia and alpha-foetoprotein detection from dried blood spots: applications to field-based studies and to clinical care in hepatitis B virus endemic areas. AB - In many resource-limited regions with endemic hepatitis B virus (HBV), there is limited infrastructure to collect, process, transport, and store blood samples for identification of persons with chronic HBV infection or with hepatocellular carcinoma (HCC). We describe the application of a simple technique using commercially available kits for detection of HBV surface antigen (HBsAg) and alpha-foetoprotein (AFP) in dried blood spots (DBS) collected on filter paper. Study participants included subjects with and without chronic HBV infection and subjects with HCC or cirrhosis. Three to five blood drops were dried on filter paper. Dried blood (equivalent to 20 muL) was eluted and tested for HBsAg by Determine(TM) HBsAg and for AFP by counter-current immuno-electrophoresis and radio-immunoassay (RIA). The primary analysis focused on comparison of DBS results to serum testing results as the gold standard. The sensitivity of DBS for detecting chronic HBV infection was 96% (98-98) with specificity of 100% (CI 99 100). Sensitivity of DBS in detecting AFP compared with serum RIA was 73% (60-86) with specificity of 90% (81-98). Both HBsAg and AFP recovery were unaffected when DBS were left at room temperature (30-33 degrees C) and under humid conditions for up to 28 days prior to elution. We conclude that DBS can be reliably used as an economical and logical alternative for detection of HBsAg in chronically infected patients and for AFP-based diagnosis of HCC in clinical situations which preclude adequate collection and processing of blood samples. Both research oriented field studies and routine clinical care may benefit from application of these techniques in resource-limited settings. PMID- 16255767 TI - Antimitochondrial antibodies in patients with chronic hepatitis C virus infection: description of 18 cases and review of the literature. AB - To describe the clinical and immunologic patterns of disease expression of patients with chronic hepatitis C virus (HCV) infection and positive antimitochondrial antibodies (AMA). We investigated the presence of AMA in 237 consecutive HCV patients with extrahepatic manifestations from an International Registry. AMA were detected by indirect immunofluorescence in triple rat tissue (liver, stomach and kidney), aceton-fixed criosections and FITC-conjugated rabbit anti-human immunoglobulins. We found positive AMA in 18 (8%) out of 237 HCV patients. All patients were female with a mean age at protocol inclusion of 65.8 years (ranging from 37 to 87 years). Twelve (67%) patients fulfilled classification criteria for systemic autoimmune diseases (SAD), including Sjogren's syndrome (n = 7), systemic sclerosis (n = 3) and systemic lupus erythematosus (n = 2). Fourteen (78%) of the HCV-AMA patients presented at least one of the highly suggestive characteristics of primary biliary cirrhosis (PBC): 9 (50%) had a specific M2 pattern, 6 (33%) had more than twice normal levels of alkaline phosphatase, 5 (28%) had raised IgM levels and 4 (22%) a histological pattern compatible with PBC. Five (28%) patients developed neoplasia after detection of AMA. Seven (39%) patients died, due to neoplasia (n = 4), cirrhotic complications (n = 2) and hepatopulmonary syndrome (n = 1). We describe a subset of HCV patients with positive AMA who presented a broad spectrum of clinical features, including liver, autoimmune and neoplasic manifestations. Two-thirds of these patients presented an associated SAD, mainly Sjogren's syndrome or systemic sclerosis, together with a high frequency of multiple autoantibodies and an increased prevalence of cirrhosis and neoplasia. PMID- 16255768 TI - Prevalence of, and risk factors for, hepatitis C virus infection among recent initiates to injecting in London and Glasgow: cross sectional analysis. AB - Our aim was to compare the prevalence of antibody to hepatitis C virus (anti-HCV) among recently initiated injecting drug users (IDUs) in London and Glasgow, and to identify risk factors which could explain differences in prevalence between the cities. Complementary studies of community recruited IDUs who had initiated injection drug use since 1996 were conducted during 2001-2002. Data on HCV risk behaviours were gathered using structured questionnaires with identical core questions and respondents were asked to provide an oral fluid specimen which was tested anonymously for anti-HCV but was linked to the questionnaire. Sensitivities of the anti-HCV assays for oral fluid were 92-96%. Prevalence of anti-HCV was 35% (122/354) in London and 57% (207/366) in Glasgow (P < 0.001). Multifactorially, factors significantly associated with raised odds of anti-HCV positivity were increasing length of injecting career, daily injection, polydrug use, having had a needlestick injury, and having served a prison sentence. In addition lower odds of anti-HCV positivity were associated with non-injection use of crack cocaine and recruitment from drug agencies. After adjustment for these factors, the increased odds of anti-HCV associated with being a Glasgow IDU were diminished but remained significant. HCV continues to be transmitted among the IDU population of both cities at high rates despite the availability of syringe exchange and methadone maintenance. Effectiveness of harm reduction interventions may be compromised by inadequate coverage and failure to reduce sufficiently the frequency of sharing different types of injecting equipment, as well as the high background prevalence of HCV, and its high infectivity. Comprehensive action is urgently required to reduce the incidence of HCV among injectors. PMID- 16255770 TI - Specificity and overlap in gene segment-defined antibody repertoires. AB - BACKGROUND: To date several studies have sought to catalog the full suite of antibodies that humans naturally produce against single antigens or other specificities (repertoire). Here we analyze the properties of all sequenced repertoires in order to better understand the specificity of antibody responses. Specifically, we ask whether the large-scale sequencing of antibody repertoires might provide a diagnostic tool for detecting antigen exposure. We do this by examining the overlap in VH-, D-, and JH- segment usage among sequenced repertoires. RESULTS: We find that repertoire overlap in VH-, D-, and JH-segment use is least for VH segments and greatest for JH segments, consistent with there being more VH than JH segments in the human genome. We find that for any two antigens chosen at random, chances are 90 percent that their repertoires' VH segments will overlap by less than half, and 98 percent that their VDJH combinations will overlap by < or =10 percent. We ran computer simulations to test whether enrichment for specific VDJH combinations could be detected in "antigen-exposed" populations, and found that enrichment is detectable with moderate-to-high sensitivity and high specificity, even when some VDJH combinations are not represented at all in some test sets. CONCLUSION: Thus, as large-scale sequencing becomes cost-effective for clinical testing, we suggest that sequencing an individual's expressed antibody repertoire has the potential to become a useful diagnostic modality. PMID- 16255771 TI - Early B-cell Factor gene association with multiple sclerosis in the Spanish population. AB - BACKGROUND: The etiology of multiple sclerosis (MS) is at present not fully elucidated, although it is considered to result from the interaction of environmental and genetic susceptibility factors. In this work we aimed at testing the Early B-cell Factor (EBF1) gene as a functional and positional candidate risk factor for this neurological disease. Axonal damage is a hallmark for multiple sclerosis clinical disability and EBF plays an evolutionarily conserved role in the expression of proteins essential for axonal pathfinding. Failure of B-cell differentiation was found in EBF-deficient mice and involvement of B-lymphocytes in MS has been suggested from their presence in cerebrospinal fluid and lesions of patients. METHODS: The role of the EBF1 gene in multiple sclerosis susceptibility was analyzed by performing a case-control study with 356 multiple sclerosis patients and 540 ethnically matched controls comparing the EBF1 polymorphism rs1368297 and the microsatellite D5S2038. RESULTS: Significant association of an EBF1-intronic polymorphism (rs1368297, A vs. T: p = 0.02; OR = 1.26 and AA vs. [TA+TT]: p = 0.02; OR = 1.39) was discovered. This association was even stronger after stratification for the well-established risk factor of multiple sclerosis in the Major Histocompatibility Complex, DRB1*1501 (AA vs. [TA+TT]: p = 0.005; OR = 1.78). A trend for association in the case-control study of another EBF1 marker, the allele 5 of the very informative microsatellite D5S2038, was corroborated by Transmission Disequilibrium Test of 53 trios (p = 0.03). CONCLUSION: Our data support EBF1 gene association with MS pathogenesis in the Spanish white population. Two genetic markers within the EBF1 gene have been found associated with this neurological disease, indicative either of their causative role or that of some other polymorphism in linkage disequilibrium with them. PMID- 16255772 TI - Treatment of dyslipidemia with lovastatin and ezetimibe in an adolescent with cholesterol ester storage disease. AB - BACKGROUND: Cholesterol ester storage disease (CESD) is an autosomal recessive illness that results from mutations in the LIPA gene encoding lysosomal acid lipase. CESD patients present in childhood with hepatomegaly and dyslipidemia characterized by elevated total and low-density lipoprotein cholesterol (LDL-C), with elevated triglycerides and depressed high-density lipoprotein cholesterol (HDL-C). Usual treatment includes a low fat diet and a statin drug. RESULTS: In an 18-year old with CESD, we documented compound heterozygosity for two LIPA mutations: a novel frameshift nonsense mutation and a deletion of exon 8. The patient had been treated with escalating doses of lovastatin for approximately 80 months, with approximately 15% decline in mean LDL-C. The addition of ezetimibe 10 mg to lovastatin 40 mg resulted in an additional approximately 16% decline in mean LDL-C. CONCLUSION: These preliminary anecdotal findings in a CESD patient with novel LIPA mutations support the longer term safety of statins in an adolescent patient and provide new data about the potential efficacy and tolerability of ezetimibe in this patient group. PMID- 16255773 TI - Changes in infant segment inertias during the first three months of independent walking. AB - BACKGROUND: During infancy, rapid changes in physical growth affect the size and shape of the body segments. To understand the effects of growth on movement, it is first necessary to quantify rates of development during the acquisition of important motor milestones. The goal of this longitudinal study was to quantify the physical growth of infant body segments during the initial stages of independent walking. METHODS: Ten infants (N = 10) aged between 28 and 55 weeks at the beginning of the study were tested biweekly (every two weeks) for three months. A 13-segment mathematical model of the human body was used to estimate the inertial parameters of the infant body segments at each session. An analysis of variance was used to test for significant differences in segment masses between biweekly measures. Polynomial contrasts were used to test for linear trends in the growth data. RESULTS: Significant differences between biweekly measures of segment mass were found only for the head/neck (F(5,45) = 3.42, p < 0.05), upper trunk (F(5,45) = 4.04, p < 0.01), and lower trunk (F(5,45) = 3.49, p < 0.01). The lower trunk demonstrated a linear increase in mass (F(1,9) = 4.56, p < 0.05). However, the upper trunk demonstrated a quadratic trend in growth (F(1,9) = 9.13, p < 0.01), while the head/neck segment showed a cubic trend in growth (F(1,9) = 3.80, p < 0.05). Significant differences in axial segment masses were also found between subjects (F(9,45) = 5.92, p < 0.001). CONCLUSION: Given that postural control proceeds in a cephalocaudal manner, the lower trunk segment would be brought under control last, in terms of the axial segments. Increases in the mass of this segment could constrain the system, thereby acting as a control parameter for the onset and development of motor patterns. PMID- 16255774 TI - Prevalence and co-occurrence of psychiatric symptom clusters in the U.S. adolescent population using DISC predictive scales. AB - OBJECTIVE: To estimate 12-month prevalence and co-occurrence of symptoms of specific mental problems among US adolescents (12-17 years) by age, sex and racial/ethnic subgroups. METHOD: Data from the 2000 National Household Survey of Drug Abuse (NHSDA) adolescent sample are used to estimate prevalence and co occurrence rates using the DISC predictive scales. Multiple logistic regressions were used to derive significant correlates of each domain of DPS-derived symptom cluster indicators of psychiatric problems and of severe comorbidity, with control of demographics and environmental factors. SETTING: The National Household Survey on Drug Abuse (NHSDA), a national household probability sample, includes a nationally representative sample of 12-17 year-old adolescents (N = 19,430), through in-home surveys. RESULTS: Three out of five adolescents screened positive for at least one DPS symptom cluster with estimates for specific symptom cluster ranging over 9.7% (substance use disorder), 13.4% (affective), 36.3% (disruptive-behavior), and 40.1% (anxiety). Co-occurrence was high with almost one-third of any DPS symptom cluster reporting multiple positive screens of four or more clusters. Blacks and younger females were most likely to report mental health problems and co-occurrence. CONCLUSION: Mental health problems among U.S. youth may be far more common than previously believed, although these symptoms have not yet reached the point of clinical impairment. The data speak to important patterns of age, gender and racial/ethnic differences in mental health problems deserving of further study. PMID- 16255775 TI - The role of surfactant protein D in the colonisation of the respiratory tract and onset of bacteraemia during pneumococcal pneumonia. AB - We have shown previously that surfactant protein D (SP-D) binds and agglutinates Streptococcus pneumoniae in vitro. In this study, the role of SP-D in innate immunity against S. pneumoniae was investigated in vivo, by comparing the outcome of intranasal infection in surfactant protein D deficient (SP-D-/-) to wildtype mice (SP-D+/+). Deficiency of SP-D was associated with enhanced colonisation and infection of the upper and lower respiratory tract and earlier onset and longer persistence of bacteraemia. Recruitment of neutrophils to inflammatory sites in the lung was similar in both strains mice in the first 24 hrs post-infection, but different by 48 hrs. T cell influx was greatly enhanced in SP-D-/- mice as compared to SP-D+/+ mice. Our data provides evidence that SP-D has a significant role to play in the clearance of pneumococci during the early stages of infection in both pulmonary sites and blood. PMID- 16255776 TI - Barriers to the implementation of programs for the prevention of mother-to-child transmission of HIV: a cross-sectional survey in rural and urban Uganda. AB - BACKGROUND: Implementation of programs for the prevention of mother-to-child transmission (PMTCT) of HIV faces a variety of barriers and challenges. The assessment of these challenges has generally been conducted in large urban health facilities. As programs expand into rural areas, the potential barriers that may be encountered there also need to be assessed. This study examines potential barriers that might affect the acceptability of interventions for PMTCT in rural and urban settings. RESULTS: Four hundred and four women at a large urban hospital and three rural clinics that had recently started implementing PMTCT were interviewed. Level of knowledge of MTCT and preference for rapid HIV testing were equally high in both areas, but rural women had a higher tendency to think that they should consult their husbands before testing, with borderline statistical significance (72% vs. 64% p = 0.09). Health facility-based deliveries were significantly lower among mothers in rural areas compared to those in the urban setting. Overall, significant predictors of willingness to test for HIV were post-primary education (OR = 3.1 95% CI 1.2, 7.7) and knowledge about rapid HIV tests (OR = 1.8, 95% CI 1.01, 3.4). The strongest predictor of willingness to accept an HIV test was the woman's perception that her husband would approve of her testing for HIV. Women who thought their husbands would approve were almost six times more likely to report a willingness to be tested compared to those who thought their husbands would not approve (OR = 5.6, 95% CI 2.8, 11.2). CONCLUSION: Lessons learned in large urban hospitals can be generalized to rural facilities, but the lower proportion of facility-based deliveries in rural areas needs to be addressed. Same-day results are likely to ensure high uptake of HIV testing services but male spousal involvement should be considered, particularly for rural areas. Universal Primary Education will support the success of PMTCT programs. PMID- 16255777 TI - Synergistic inhibition of human melanoma proliferation by combination treatment with B-Raf inhibitor BAY43-9006 and mTOR inhibitor Rapamycin. AB - BACKGROUND: Targeted inhibition of protein kinases is now acknowledged as an effective approach for cancer therapy. However, targeted therapies probably have limited success because cancer cells have alternate pathways for survival and proliferation thereby avoiding inhibition. We tested the hypothesis that combination of targeted agents would be more effective than single agents in arresting melanoma cell proliferation. METHODS: We evaluated whether BAY43-9006, an inhibitor of the B-Raf kinase, and rapamycin, an inhibitor of the mTOR kinase, would inhibit serum-stimulated proliferation of human melanoma cell lines, either alone or in combination. Proliferation was measured by quantitating melanoma cell numbers with a luciferase for ATP. Phosphorylation of proteins downstream of targeted kinase(s) was assayed by immunoblots. Statistical significance was determined with the Student-T test. Isobologram analysis was performed to distinguish additive versus synergistic effects of combinations of drugs. RESULTS: Serum-stimulated proliferation of multiple human melanoma cell lines was inhibited by BAY43-9006 and by rapamycin. Melanoma cells containing the B-Raf mutation V599E were more sensitive than cells with wild-type B-raf to 10 nM doses of both BAY43-9006 and rapamycin. Regardless of B-Raf mutational status, the combination of low dose rapamycin and BAY43-9006 synergistically inhibited melanoma cell proliferation. As expected, rapamycin inhibited the phosphorylation of mTOR substrates, p70S6K and 4EBP1, and BAY43-9006 inhibited phosphorylation of ERK, which is dependent on B-Raf activity. We also observed unexpected rapamycin inhibition of the phosphorylation of ERK, as well as BAY43-9006 inhibition of the phosphorylation of mTOR substrates, p70S6K and 4EBP1. CONCLUSION: There was synergistic inhibition of melanoma cell proliferation by the combination of rapamycin and BAY 43-9006, and unexpected inhibition of two signaling pathways by agents thought to target only one of those pathways. These results indicate that combinations of inhibitors of mTOR and of the B-raf signaling pathways may be more effective as a treatment for melanoma than use of either agent alone. PMID- 16255779 TI - Time course of exercise induced alterations in daily activity in chronic fatigue syndrome. AB - In a previous study we demonstrated that while people with CFS had lower daily activity levels than control subjects, they were able to increase daily activity via a daily walking program. We reanalyzed our data to determine the time course of activity changes during the walking program. Daily activity assessed via an accelometer worn at the hip was divided into sleep, active, and walking periods. Over the first 4-10 days of walking the subjects with CFS were able to reach the prescribed activity goals each day. After this time, walking and total activity counts decreased. Sedentary controls subjects were able to maintain their daily walking and total activity goals throughout the 4 weeks. Unlike our previous interpretation of the data, we feel this new analysis suggests that CFS patients may develop exercise intolerance as demonstrated by reduced total activity after 4-10 days. The inability to sustain target activity levels, associated with pronounced worsening of symptomology, suggests the subjects with CFS had reached their activity limit. PMID- 16255780 TI - Compliance of a cobalt chromium coronary stent alloy--the COVIS trial. AB - BACKGROUND: Cobalt chromium coronary stents are increasingly being used in percutaneous coronary interventions. There are, however, no reliable data about the characteristics of unfolding and visibility of this stent alloy in vivo. The aim of this study is to compare cobalt chromium coronary stents with conventional stainless steel stents using intracoronary ultrasound. METHODS: Twenty de novo native coronary stenoses < or = 20 mm in length (target vessel reference diameter > or = 2.5 and < or = 4.0 mm) received under sequential intracoronary ultrasound either a cobalt chromium stent (Multi-Link Vision; n = 10) or a stainless steel stent (Multi-Link Zeta; n = 10). RESULTS: For optimal unfolding, the cobalt chromium stent requires a higher balloon deployment pressure (13.90 +/- 2.03 atm) than the stainless steel stent (11.50 +/- 2.12 atm). Furthermore, the achieved target vessel diameter of the cobalt chromium stent (Visibility-Index QCA/IVUS Multi-Link Vision 1.13 / Multi-Link Zeta 1.04) is more easily overrated by Quantitative Coronary Analysis. CONCLUSION: These data indicate that stent material-specific recommendations for optimal implantation pressure and different stent material with an equal design should both be considered in interpreting QCA analysis. PMID- 16255778 TI - Antimetastatic gene expression profiles mediated by retinoic acid receptor beta 2 in MDA-MB-435 breast cancer cells. AB - BACKGROUND: The retinoic acid receptor beta 2 (RARbeta2) gene modulates proliferation and survival of cultured human breast cancer cells. Previously we showed that ectopic expression of RARbeta2 in a mouse xenograft model prevented metastasis, even in the absence of the ligand, all-trans retinoic acid. We investigated both cultured cells and xenograft tumors in order to delineate the gene expression profiles responsible for an antimetastatic phenotype. METHODS: RNA from MDA-MB-435 human breast cancer cells transduced with RARbeta2 or empty retroviral vector (LXSN) was analyzed using Agilent Human 1A Oligo microarrays. The one hundred probes with the greatest differential intensity (p < 0.004, jointly) were determined by selecting the top median log ratios from eight-paired microarrays. Validation of differences in expression was done using Northern blot analysis and quantitative RT-PCR (qRT-PCR). We determined expression of selected genes in xenograft tumors. RESULTS: RARbeta2 cells exhibit gene profiles with overrepresentation of genes from Xq28 (p = 2 x 10(-8)), a cytogenetic region that contains a large portion of the cancer/testis antigen gene family. Other functions or factors impacted by the presence of exogenous RARbeta2 include mediators of the immune response and transcriptional regulatory mechanisms. Thirteen of fifteen (87%) of the genes evaluated in xenograft tumors were consistent with differences we found in the cell cultures (p = 0.007). CONCLUSION: Antimetastatic RARbeta2 signalling, direct or indirect, results in an elevation of expression for genes such as tumor-cell antigens (CTAG1 and CTAG2), those involved in innate immune response (e.g., RIG-I/DDX58), and tumor suppressor functions (e.g., TYRP1). Genes whose expression is diminished by RARbeta2 signalling include cell adhesion functions (e.g, CD164) nutritional or metabolic processes (e.g., FABP6), and the transcription factor, JUN. PMID- 16255781 TI - Assessment of soy phytoestrogens' effects on bone turnover indicators in menopausal women with osteopenia in Iran: a before and after clinical trial. AB - BACKGROUND: Osteoporosis is the gradual declining in bone mass with age, leading to increased bone fragility and fractures. Fractures in hip and spine are known to be the most important complication of the disease which leads in the annual mortality rate of 20% and serious morbidity rate of 50%. Menopause is one of the most common risk factors of osteoporosis. After menopause, sex hormone deficiency is associated with increased remodeling rate and negative bone balance, leading to accelerated bone loss and micro-architectural defects, resulting into increased bone fragility. Compounds with estrogen-like biological activity similar to "Isoflavones" present in plants especially soy, may reduce bone loss in postmenopausal women as they are similar in structure to estrogens. This research, therefore, was carried out to study the effects of Iranian soy protein on biochemical indicators of bone metabolism in osteopenic menopausal women. MATERIALS AND METHODS: This clinical trial of before-after type was carried out on 15 women 45-64 years of age. Subjects were given 35 g soy protein per day for 12 weeks. Blood and urine sampling, anthropometric measurement and 48-h-dietary recalls were carried out at zero, 6 and 12 weeks. Food consumption data were analyzed using Food Proccessor Software. For the study of bone metabolism indicators and changes in anthropometric data as well as dietary intake, and repeated analyses were employed. RESULTS: Comparison of weight, BMI, physical activity, energy intake and other intervening nutrients did not reveal any significant changes during different stages of the study. Soy protein consumption resulted in a significant reduction in the urinary deoxypyridinoline and increasing of total alkaline phosphatase (p < 0.05), although the alterations in osteocalcin, c-telopeptide, IGFBP3 and type I collagen telopeptide were not significant. CONCLUSION: In view of beneficial effect of soy protein on bone metabolism indicators, inclusion of this relatively inexpensive food in the daily diet of menopausal women, will probably delay bone resorption, thereby preventing osteoporosis. PMID- 16255782 TI - CEBPG transcription factor correlates with antioxidant and DNA repair genes in normal bronchial epithelial cells but not in individuals with bronchogenic carcinoma. AB - BACKGROUND: Cigarette smoking is the primary cause of bronchogenic carcinoma (BC), yet only 10-15% of heavy smokers develop BC and it is likely that this variation in risk is, in part, genetically determined. We previously reported a set of antioxidant genes for which transcript abundance was lower in normal bronchial epithelial cells (NBEC) of BC individuals compared to non-BC individuals. In unpublished studies of the same NBEC samples, transcript abundance values for several DNA repair genes were correlated with these antioxidant genes. From these data, we hypothesized that antioxidant and DNA repair genes are co-regulated by one or more transcription factors and that inter individual variation in expression and/or function of one or more of these transcription factors is responsible for inter-individual variation in risk for BC. METHODS: The putative transcription factor recognition sites common to six of the antioxidant genes were identified through in silico DNA sequence analysis. The transcript abundance values of these transcription factors (n = 6) and an expanded group of antioxidant and DNA repair genes (n = 16) were measured simultaneously by quantitative PCR in NBEC of 24 non-BC and 25 BC individuals. RESULTS: CEBPG transcription factor was significantly (p < 0.01) correlated with eight of the antioxidant or DNA repair genes in non-BC individuals but not in BC individuals. In BC individuals the correlation with CEBPG was significantly (p < 0.01) lower than that of non-BC individuals for four of the genes (XRCC1, ERCC5, GSTP1, and SOD1) and the difference was nearly significant for GPX1. The only other transcription factor correlated with any of these five target genes in non BC individuals was E2F1. E2F1 was correlated with GSTP1 among non-BC individuals, but in contrast to CEBPG, there was no significant difference in this correlation in non-BC individuals compared to BC individuals. CONCLUSION: We conclude that CEBPG is the transcription factor primarily responsible for regulating transcription of key antioxidant and DNA repair genes in non-BC individuals. Further, we conclude that the heavy smokers selected for development of BC are those who have sub-optimal regulation of antioxidant and DNA repair genes by CEBPG. PMID- 16255783 TI - The efficiency of different search strategies in estimating parsimony jackknife, bootstrap, and Bremer support. AB - BACKGROUND: For parsimony analyses, the most common way to estimate confidence is by resampling plans (nonparametric bootstrap, jackknife), and Bremer support (Decay indices). The recent literature reveals that parameter settings that are quite commonly employed are not those that are recommended by theoretical considerations and by previous empirical studies. The optimal search strategy to be applied during resampling was previously addressed solely via standard search strategies available in PAUP*. The question of a compromise between search extensiveness and improved support accuracy for Bremer support received even less attention. A set of experiments was conducted on different datasets to find an empirical cut-off point at which increased search extensiveness does not significantly change Bremer support and jackknife or bootstrap proportions any more. RESULTS: For the number of replicates needed for accurate estimates of support in resampling plans, a diagram is provided that helps to address the question whether apparently different support values really differ significantly. It is shown that the use of random addition cycles and parsimony ratchet iterations during bootstrapping does not translate into higher support, nor does any extension of the search extensiveness beyond the rather moderate effort of TBR (tree bisection and reconnection branch swapping) plus saving one tree per replicate. Instead, in case of very large matrices, saving more than one shortest tree per iteration and using a strict consensus tree of these yields decreased support compared to saving only one tree. This can be interpreted as a small risk of overestimating support but should be more than compensated by other factors that counteract an enhanced type I error. With regard to Bremer support, a rule of thumb can be derived stating that not much is gained relative to the surplus computational effort when searches are extended beyond 20 ratchet iterations per constrained node, at least not for datasets that fall within the size range found in the current literature. CONCLUSION: In view of these results, calculating bootstrap or jackknife proportions with narrow confidence intervals even for very large datasets can be achieved with less expense than often thought. In particular, iterated bootstrap methods that aim at reducing statistical bias inherent to these proportions are more feasible when the individual bootstrap searches require less time. PMID- 16255784 TI - Pulmonary function and fuel use: a population survey. AB - BACKGROUND: In the backdrop of conflicting reports (some studies reported adverse outcomes of biomass fuel use whereas few studies reported absence of any association between adverse health effect and fuel use, may be due to presence of large number of confounding variables) on the respiratory health effects of biomass fuel use, this cross sectional survey was undertaken to understand the role of fuel use on pulmonary function. METHOD: This study was conducted in a village of western India involving 369 randomly selected adult subjects (165 male and 204 female). All the subjects were interviewed and were subjected to pulmonary function test. Analysis of covariance was performed to compare the levels of different pulmonary function test parameters in relation to different fuel use taking care of the role of possible confounding factors. RESULTS: This study showed that biomass fuel use (especially wood) is an important factor for deterioration of pulmonary function (particularly in female). FEV1 (p < .05), FEV1% (p < .01), PEFR (p < .05) and FEF(25-75) (p < .01) values were significantly lower in biomass fuel using females than nonusers. Comparison of only biomass fuel use vs. only LPG (Liquefied Petroleum Gas) use and only wood vs. only LPG use has showed that LPG is a safer fuel so far as deterioration of pulmonary function is concerned. This study observes some deterioration of pulmonary function in the male subjects also, who came from biomass fuel using families. CONCLUSION: This study concluded that traditional biomass fuels like wood have adverse effects on pulmonary function. PMID- 16255785 TI - A comparative study of food habits and body shape perception of university students in Japan and Korea. AB - BACKGROUND: Abnormal body weight, dietary concerns, and unhealthy weight loss behaviors are increasingly being observed in young females in Japan. Our previous research has shown that the irregular lifestyles of female Japanese and Chinese students are significantly related to their desire to be thinner. In the present study, we compare the food habits and body shape preferences of female university students in South Korea and Japan to explore body shape perceptions in those populations. METHODS: A total of 265 female university students aged 19-25 years participated in this study. University students in Korea (n = 141) and university students in Japan (n = 124) completed a self-reported questionnaire. Data were analyzed using SPSS statistical software. Descriptive statistics were used to identify the demographic characteristics of the students and parametric variables were analyzed using the Student's t-test. Chi-square analyses were conducted for non-parametric variables. RESULTS: Comparison of body mass index (BMI) distributions in Japan and Korea showed the highest value in the normal category (74%) together with a very low obesity rate (1.2%). Significant differences were observed between the two countries in terms of eating patterns, with more Japanese eating breakfast daily and with Japanese students eating meals more regularly than Korean students. A difference was also observed in frequency of meals, where Korean students reported eating meals two times per day (59%) and the majority of Japanese students reported eating meals three times per day (81%). Although most subjects belonged to the normal BMI category, their ideal BMI classification was the underweight category (BMI: 18.4 +/- 3.4). CONCLUSION: Few studies have compared the health related practices of Japanese and Korean university students. The present results suggest the necessity of nutrition and health promotion programs for university students, especially programs emphasizing weight management. PMID- 16255787 TI - Hand-assisted right laparoscopic live donor nephrectomy. AB - PURPOSE: Laparoscopic live donor nephrectomy has acquired an important role in the era of minimally invasive surgery. Laparoscopic harvesting of the right kidney is technically more challenging than that of the left kidney because of the short right renal vein and the need to retract the liver away from the right kidney. The aim of this article is to report our experience with right laparoscopic live donor nephrectomies. MATERIALS AND METHODS: We performed a retrospective review of 28 patients who underwent right laparoscopic donor nephrectomies at our service. Operative data and postoperative outcomes were collected, including surgical time, estimated blood loss, warm ischemia time, length of hospital stay, conversion to laparotomy and complications. RESULTS: The procedure was performed successfully in all 28 patients. The mean operative time was 83.8 minutes (range 45 to 180 minutes), with an estimated blood loss of 111.4 mL (range 40 to 350 mL) and warm ischemia time of 3 minutes (range 1.5 to 8 minutes). No donor needed conversion to open surgery and all kidneys showed immediate function after implantation. The average time to initial fluid intake was 12 hours (range 8 to 24 hours). Two cases of postoperative ileus and a case of hematoma on the hand-port site were observed. The mean postoperative hospital stay was 3 days (range 1 to 7 days). CONCLUSIONS: Our data confirm the safety and feasibility of right laparoscopic donor nephrectomy and we believe that the right kidney should not be avoided for laparoscopic donor nephrectomy when indicated. PMID- 16255788 TI - Likelihood of retrograde double-J stenting according to ureteral obstructing pathology. AB - OBJECTIVES: To evaluate the likelihood of retrograde double-J stenting in urgent ureteral drainage according to obstructing pathology. MATERIALS AND METHODS: From July 2002 to January 2003, 43 consecutive patients with ureteral obstruction who needed urgent decompression were evaluated at our institution, where we performed a total of 47 procedures. Emergency was defined as ureteral obstruction associated with infection, obstructive acute renal failure, or refractory pain. Ureteral obstruction was defined as intrinsic and extrinsic based on etiology and evaluated by ultrasound. Patients submitted to previous double-J stenting were excluded. Failures in retrograde ureteral stenting were treated with percutaneous nephrostomy. Results were analyzed with Fisher's exact test and regression analysis. RESULTS: Failure in retrograde ureteral stenting occurred in 9% (2/22) and 52% (13/25) of the attempts in patients with intrinsic and extrinsic obstruction respectively (p < 0.001). Failures in stenting extrinsic obstructions occurred due to lack of identification of the ureteral meatus in 77% and impossibility of catheter progression in 23% (p < 0.05). All attempts of retrograde catheter insertion failed in obstructions caused by prostate or bladder pathologies (6/6). Inability to identify the ureteral meatus was the cause of all failures. CONCLUSION: Retrograde double-J stenting has a low probability of success in extrinsic ureteral obstruction caused by prostate or bladder disease. Such cases might be best managed with percutaneous nephrostomy. PMID- 16255789 TI - Serum PSA and cure perspective for prostate cancer in males with nonpalpable tumor. AB - INTRODUCTION: Many studies have shown the association between PSA levels and the subsequent detection of prostate cancer. In the present trial, we have studied the relationship between preoperative PSA levels and clinical outcome following radical prostatectomy in men with clinical stage T1c. MATERIALS AND METHODS: 257 individuals with clinical stage T1c undergoing retropubic radical prostatectomy were selected in the period from 1991 to 2000. Following surgery, biochemical recurrence-free survival curves were constructed according to PSA levels between 0-4; 4.1-10; 10.1-20 and > 20 ng/mL. RESULTS: Of the total of 257 selected patients, 206 (80%) had Gleason scores from 2 to 6 and 51 (20%), presented Gleason scores 7 and 8, as defined by the pathological report from prostate biopsy. There was no biochemical recurrence of disease when the PSA was lower than 4, regardless of Gleason score. Biochemical recurrence-free survival according to PSA between 0-4; 4.1-10; 10.1-20 and > 20 was 100%, 87.6%, 79% and 68.8% for Gleason scores 2-6 and 100%; 79.4%; 40% and 100% for Gleason scores 7-8 respectively. When all individuals were grouped, regardless of their Gleason scores, the probability of biochemical recurrence-free survival was 100%, 65.1%, 53.4% and 72.2% according to PSA between 0-4; 4.1-10; 10.1-20 and > 20 ng/mL respectively. CONCLUSION: Non-palpable prostate cancer presents higher chances of cure when the PSA is inferior to 4 ng/mL. PMID- 16255790 TI - Prediction of pathological stage in prostate cancer through the percentage of involved fragments upon biopsy. AB - INTRODUCTION: The need for defining the extension of disease in patients undergoing radical prostatectomy due to prostate adenocarcinoma is a relevant factor cure in such individuals. In order to identify a new independent preoperative factor for predicting the extension of prostate cancer, we assessed the role of the percentage of positive fragments upon biopsy. MATERIALS AND METHODS: A retrospective study compared the percentage of positive fragments on biopsy with the extension of disease as defined by the pathological examination of the surgical specimen from 898 patients undergoing radical prostatectomy due to clinically localized prostate cancer. RESULTS: On the univariate analysis, the percentage of positive fragments on biopsy showed a statistical significance for predicting confined disease (p < 0.001), which was found in 66.7% of the cases under study. Additionally, we observed that the total number of removed fragments exerts no influence on the extension of the disease (p = 0.567). CONCLUSION: the percentage of positive fragments is an independent factor for predicting the pathological stage of prostate adenocarcinoma, and the number of removed fragments is not related to the extension of the disease. PMID- 16255791 TI - Assessment of sexual function in patients undergoing vasectomy using the international index of erectile function. AB - INTRODUCTION: The present study aims to prospectively compare the sexual function in males before and after vasectomy surgery using the international index of erectile function (IIEF). MATERIALS AND METHODS: From October to December 2002, sixty-four patients who were candidates for male sterilization in the vasectomy program of the Urology Section at the General Hospital of the University of Sao Paulo were included. The same investigator applied the IIEF before and 90 days after the surgery. The mean scores obtained on pre and postoperative visits for all domains of sexual function were analyzed and compared with the Wilcoxon test. RESULTS: The mean patient age was 35 years (range from 25 to 48 years) and the mean number of children per man was 3. The total mean score of the IIEF was 64.06 before surgery and 65.64 after the procedure, with this difference considered statistically significant (p < 0.001). Sixty-seven per cent of the patients improved their scores, versus 17% and 16% who showed worsening or no change at all in IIEF scores following surgery, respectively. Of the 5 sexual function domains, desire and sexual satisfaction presented statistically significant improvement. CONCLUSION: This study showed that vasectomy caused a positive impact on sexual function, especially on desire and sexual satisfaction, in the majority of men undergoing surgery. There was no case of surgery-related erectile dysfunction. PMID- 16255792 TI - Early catheter removal after anterior anastomotic (3 days) and ventral buccal mucosal onlay (7 days) urethroplasty. AB - INTRODUCTION: Physicians who perform urethroplasty have varying opinions about when the urinary catheter should be removed post-operatively, but research on this subject has not yet appeared in the literature. We performed voiding cystourethrogram (VCUG) on our anterior urethroplasty patients on days 3 (anastomotic) and 7 (buccal) in an effort to determine the earliest day for removal of the urethral catheter. MATERIALS AND METHODS: Retrospective chart review of 29 urethroplasty patients from October 2002-August 2004 was performed at two reconstructive urology centers. 17 patients had early catheter removal (12 anastomotic and 5 ventral buccal onlay urethroplasty) and were compared to 12 who had late removal (7 anastomotic and 5 buccal). RESULTS: Of those with early catheter removal, 2/12 (17%) of anastomotic urethroplasty patients had extravasation, which resolved by the following week and 0/5 (0%) of the buccal mucosal urethroplasty patients had extravasation. Patients with late catheter removal underwent VCUG 6-14 days (mean 8 days) after anastomotic urethroplasty and 9-14 days (mean 12 days) after buccal mucosal urethroplasty. 0% of the anastomotic urethroplasty had leakage after the late VCUG and 1/5 (20%) of the buccal patients had extravasation after the VCUG. Recurrences were low in all patient groups. CONCLUSION: Catheter removal after anastomotic and buccal mucosal urethroplasty can be safely attempted on the 3rd and 7th post-operative days respectively, with a low rate of extravasation on VCUG. Eliminating the catheter as soon as possible should improve patient comfort without harming results and decrease the overall negative impact of surgery on the patient. PMID- 16255793 TI - Collecting duct carcinoma associated with oncocytoma. AB - Collecting duct carcinoma (CDC) is a rare, highly aggressive malignant neoplasm that arises from the collecting duct epithelium of the kidney. CDC was reported to coexist with renal cell and transitional cell carcinomas. We report a rare case of CDC associated with oncocytoma, confirmed by the characteristic histological appearance and immunohistochemistry. We also review the epidemiological, histological and immunohistochemical criteria for diagnosis, in addition to the genetic and cytogenetic aberrations reported in the literature. Identification and reporting CDC is important for the establishment of treatment strategies and monitoring prognosis. PMID- 16255794 TI - Right atrial migration of nephrostomy catheter. AB - Percutaneous tube nephrostomy (PTN) placement is associated with bleeding complications in a small proportion of cases. We study a case of inadvertent renal vein catheterization during PTN tube change with catheter right atrial migration treated by fluoroscopically monitored catheter removal. PMID- 16255795 TI - Intravesical wire as foreign body in urinary bladder. AB - Foreign bodies in the urinary bladder are frequently the objects of jokes among doctors, but they may sometimes cause serious implications to the patients. Here we present our experiences in 3 such cases where long segments of wire were introduced into the urinary bladder through the urethra. PMID- 16255797 TI - Early diagnosis of the urofacial syndrome is essential to prevent irreversible renal failure. AB - INTRODUCTION: The urofacial or Ochoa syndrome is a rare disease characterized by the presence of functional obstructive uropathy associated with peculiar facial features when patients attempt to smile or laugh. Unfortunately, many of these patients remain without proper diagnosis or adequate treatment due to lack of recognition of the disease. This can ultimately result in upper tract deterioration and eventual renal failure. We present our experience with this rare syndrome. MATERIALS AND METHODS: We identified 3 patients who presented initially with acute renal failure, urinary tract infection (UTI) and severe dysfunctional elimination. All patients were thoroughly evaluated, including screening for spinal cord anomalies, and were subsequently diagnosed with urofacial syndrome. RESULTS: At the outset, the two older patients (aged 4 and 9 years) presented with the typical facial features when attempting to smile or laugh. One patient in the newborn period presented with urinary and fecal retention and septicemia and, to our knowledge, represents the youngest case of urofacial syndrome reported so far. All patients were evaluated with ultrasonography, renal scan, voiding cystourethrogram (VCUG) and urodynamics. Findings included hydronephrosis and a thick-walled, trabeculated bladder with poor compliance and detrusor hypereflexia respectively in each patient. All were subsequently treated with clean intermittent catheterization (CIC), antibiotic prophylaxis and anticholinergic therapy. One patient required appendicovesicostomy for CIC due to discomfort secondary to a sensate urethra. CONCLUSIONS: Our series demonstrates that early recognition of this rare syndrome is necessary to adequately treat and prevent upper tract deterioration in these unique individuals. Although the urofacial is difficult to diagnose in infants, cognizance must be maintained in order to prevent severe subsequent sequalae. PMID- 16255796 TI - Aggressive vaginal angiomyxoma mimicking urethral tumor. AB - This is a case report of a 32-year-old female patient with a neoplasia mimicking a urethral tumor. Following anterior pelvic exanteration, vulvectomy, bilateral inguinal lymphadenectomy, the pathological study established the diagnosis of aggressive vaginal angiomyxoma, CD-34 labeled. PMID- 16255798 TI - Responsiveness to the Portuguese version of the International Consultation on Incontinence Questionnaire-Short Form (ICIQ-SF) after stress urinary incontinence surgery. AB - OBJECTIVE: To evaluate the reliability and responsiveness (internal and external) of the Portuguese version of the ICIQ-SF. We assessed the responsiveness of the ICIQ-SF after surgical procedures for the treatment of stress urinary incontinence. MATERIALS AND METHODS: Prospective open label study in 2 tertiary referral centers. Sixty-one patients of both genders (54 female and 7 male) were enrolled. Patients were treated using surgical procedures, mostly with synthetic sling (82%). Patients were assessed before surgery and at least 1 month postoperatively using the ICIQ-SF in its translated and validated Portuguese version. Patients also underwent pre-operative urodynamic tests, Stamey incontinence grading and pad usage assessments. After surgery, patients underwent stress tests, Stamey incontinence grading and pad usage assessments. RESULTS: The mean age was 57.2 (+/- 11.6) years and the mean duration of follow-up was 7.2 months (+/- 4.5). Objective parameters such as urodynamic tests (by means of VLPP) and pad usage had significant correlation with changes in post-treatment scores on the ICIQ-SF (p = 0.0062 and p < 0.0001 respectively). The responsiveness expressed in terms of standardized effect sizes (SES) and standardized response means (SRM) was large for both questionnaires (p < 0.0001). CONCLUSION: The results showed high responsiveness (large effect sizes I and II) for the Portuguese version of the ICIQ-SF, indicating that this instrument is suitable for measuring outcomes in clinical trials for Brazilian patients with stress urinary incontinence. PMID- 16255816 TI - Attachment and invasion of Toxoplasma gondii and Neospora caninum to epithelial and fibroblast cell lines in vitro. AB - Attachment and invasion of Toxoplasma gondii and Neospora caninum to a cat and a dog fibroblast cell line and 2 epithelial cell lines (a cat kidney and Vero) were compared in vitro using fluorescence antibody methodology. In addition, trypsin treatment of tachyzoites was used to determine whether protein molecules were essential to the process of invasion. The results show that both T. gondii and N. caninum invaded all 4 cell lines, and that pre-treatment of T. gondii tachyzoites with trypsin caused an increase in the ability of the parasite to invade these host cells. Furthermore T. gondii, in comparison to N. caninum, invaded all 4 cell lines at greater levels. The results here support the conclusion that both T. gondii and N. caninum have the ability to invade a variety of cell types including both dog and cat cells, and questions the utility of Vero cells as an appropriate host cell for in vitro studies on the biology of these taxa. PMID- 16255817 TI - A small ciliary surface glycoprotein of the monogenean parasite Neobenedenia girellae acts as an agglutination/immobilization antigen and induces an immune response in the Japanese flounder Paralichthys olivaceus. AB - The capsalid monogenean Neobenedenia girellae, a parasite of seawater fishes, was found to express an antigen that elicits antibodies in rabbits, and these antibodies had agglutination/immobilization activity against N. girellae larvae (oncomiracidia) in vitro. Indirect immunofluorescence staining of N. girellae oncomiracidia showed that this agglutination/immobilization antigen was expressed on the surface of cilia. An intraperitoneal injection of ciliary proteins (either sonicated or intact) with adjuvant also elicited agglutinizing/immobilizing antibodies in sera from Japanese flounder, Paralichthys olivaceus. These antisera showed a clear correlation between anti-ciliary antibody levels (measured by enzyme-linked immunosorbent assays) and their agglutination/immobilization activity. Anti-ciliary antibody levels in Japanese flounder reached a plateau at 39 days after booster immunization and were significantly higher in the two immunized groups (injection of sonicated or intact cilia) as compared with control fish (injection of bovine serum albumin; ANOVA, Tukey's test, P < 0.01). Anti-ciliary antibodies were also found in fish mucus; however, there was no correlation between fish serum anti-ciliary antibody levels and mucus antibody levels. A Triton X-114-soluble 8 kDa glycoprotein of the ciliary integral membrane fraction is a plausible candidate for the agglutination/immobilization antigen based on SDS-polyacrylamide gel electrophoresis and immunoblot analyses with rabbit and fish antisera. PMID- 16255818 TI - Stimulation of innate immunity in newborn kids against Cryptosporidium parvum infection-challenge by intranasal/per-oral administration of liposomal formulation of N-L18-norAbu-GMDP adjuvant. AB - The effects of a liposomal preparation of lipophilic immunomodulator beta-D GlcNstearoyl-(1-4)-norMurNAc-L-Abu-D-isoGln (N-L18-norAbu-GMDP) were investigated on resistance to Cryptosporidium parvum infection in neonatal kids. The liposomal preparation was administered subcutaneously or intranasally/orally (i.n./p.o.) twice at doses of 100 microg, 200 microg, or 1000 microg per kid pre-infection challenge. The treatment schemes were (i) 72 and 24 h pre-infection challenge, (ii) 24 h pre-infection challenge and 24 h post-infection challenge (oral inoculation with 1 x 10(7) oocysts of C. parvum in 5 ml of PBS). Administration of liposomal N-L18-norAbu-GMDP by i.n./p.o. route at the cumulative dose of 2000 microg per kid 72 and 24 h pre-infection challenge, lead to substantially increased clearance of coccidian parasites from various parts of the intestine. On the basis of histological examination, the distribution of cryptosporidia in the intestine and the severity of the infection, treated kids were classified on day 5 as having a strong reduction in infection in comparison to the control group (P < 0.05). No cryptosporidia were found on the mucosal surface of treated kids by day 10, while the intestines of the control kids were still infected. All doses and routes of administration were judged effective with respect to suppression of cryptosporidia infections. PMID- 16255819 TI - Polar filament discharge of Myxobolus cerebralis actinospores is triggered by combined non-specific mechanical and chemical cues. AB - This study presents initial evidence for the requirement of both chemical and mechanical stimuli to discharge polar capsules of Myxobolus cerebralis actinospores, the causative agent of salmonid whirling disease. The obligate need for combined discharge triggers was concluded from data obtained in a before/after experimental set-up carried out with individual locally immobilized actinospores. Homogenized rainbow trout mucus as chemostimulus and tangency of the apical region of the spores to achieve mechanical stimulation were applied subsequently. The actinospores showed discharged polar filaments exclusively when mucus substrate application was followed by touching the polar capsule-bearing region, but not when either stimulus was offered solely to the same individuals. We measured filament discharge rates to mucus preparations in a microscopic assay using supplementary vibration stimuli to ensure mechanical excitation. The actinospores responded similarly to different frequencies, which suggested a touch-sensitive recognition mechanism. Discharge specificity for salmonid mucus could not be confirmed, as mucus of common carp and bream could trigger similar filament expulsion rates. To a lesser extent homogenized frog epidermis and bovine submaxillary mucin could also stimulate the attachment reaction. In contrast, mucus of a pulmonate freshwater snail elicited no response. PMID- 16255821 TI - Comparative study of the presence of Trypanosoma cruzi kDNA, inflammation and denervation in chagasic patients with and without megaesophagus. AB - Neuronal lesions have been considered the hallmark of chagasic megaesophagus, but the role of Trypanosoma cruzi and the participation of the inflammatory cells in this process are still debated. In the present study we counted neurons in the oesophagus from patients with and without megaesophagus and further examined these samples for the presence of parasite kDNA and cells with cytolytic potential (Natural Killer cells, cytotoxic lymphocytes and macrophages). The presence of parasite kDNA was demonstrated in 100% of cases with megaesophagus and in 60% of patients without megaesophagus. When analysed for the number of neurons, the patients without megaesophagus could be classified into 2 groups, as having normal or a decreased number of neurons. The former group did not show any inflammatory process, but interestingly, all patients without megaesophagus presenting decreased number of neurons also presented both parasite kDNA and inflammatory process in the organ. We further observed that the numbers of cytotoxic cells in the myenteric plexus region inversely correlate with the number of neurons. These data together strongly suggest that chronic lesions in chagasic megaesophagus might be a consequence of immune-mediated mechanisms, that last until the chronic phase of infection, and are dependent on the persistence of parasite in the host's tissue. PMID- 16255820 TI - Population genetics of multi-host parasites--the case for molecular epidemiological studies of Schistosoma japonicum using larval stages from naturally infected hosts. AB - Population genetics of multi-host pathogens offers great potential for the understanding of their complex epidemiology but care must be taken to ensure that the sampling procedure does not bias estimates of population indices. The transfer of material to laboratory passage, in particular, runs the risk of bottlenecking and imposing non-random host-induced selection pressures according to the hosts used in passage. We present a novel technique allowing single-locus microsatellite genotyping of the naturally sampled larval stages, enabling unbiased population genetic studies of the multi-host zoonotic parasite Schistosoma japonicum. The utility of these larval genotyping methods for molecular epidemiological studies are illustrated in results from 3 separate data sets. In the first data set, potential loss of alleles based on the definitive host species used for laboratory maintenance was identified by comparing adult worm populations derived from mice and rabbits infected with cercarial populations originating from the same set of snails. In the second data set, bottlenecking was demonstrated by the loss of alleles in adult worms derived within a single generation of laboratory maintenance compared to their parent field-collected cercarial samples. In the final data set, comparison of miracidia and adult worms recovered from naturally infected animals demonstrated that larval analyses can provide stage-specific epidemiological information and that population genetics of schistosomes can be well described by analysis of larval stages. Our results thus advocate the use of natural life-cycle stages to obtain an accurate and ethical representation of the population genetic structure of S. japonicum and other multi-host pathogens. PMID- 16255822 TI - Acquired resistance to berenil in a cloned isolate of Trypanosoma evansi is associated with upregulation of a novel gene, TeDR40. AB - Drug resistance is now a severe and increasing problem in trypanosomes, but molecular details of mechanisms of resistance are only beginning to unveil. There is urgent need to clearly elucidate the different mechanisms of drug resistance in trypanosomes in order to circumvent existing resistance problems and avoid emergence of resistance to the next generation drugs. In this study, we cloned and characterized a novel gene, TeDR40, whose expression is associated with resistance to berenil in Trypanosoma evansi. Expression analysis showed that the gene was at least 1000-fold upregulated in resistant parasites and the encoded protein appeared to have a ubiquitous cellular localization. To investigate the association of TeDR40 with berenil-resistance, we genetically modified wild-type berenil-sensitive T. evansi for inducible over-expression of the TeDR40 gene. Induction of over-expression of TeDR40 in T. evansi led to decreased (P < 0.01) sensitivity to berenil. Our findings indicate a possible correlation between over expression of a novel gene, TeDR40, and reduced sensitivity to berenil in an in vitro-cultured clonal line of T. evansi. PMID- 16255823 TI - Delayed transmission of a parasite is compensated by accelerated growth. AB - Compensatory or 'catch-up' growth following prolonged periods of food shortages is known to exist in many free-living animals. It is generally assumed that growth rates under normal circumstances are below maximum because elevated rates of growth are costly. The present paper gives experimental evidence that such compensatory growth mechanisms also exist in parasitic species. We explored the effect of periodic host unavailability on survival, infectivity and growth of the fish ectoparasite Argulus coregoni. Survival and infectivity of A. coregoni metanauplii deprived of a host for selected time periods were age dependent, which indicates that all metanauplii carry similar energy resources for host seeking. Following the periods off-host, metanauplii were allowed to settle on rainbow trout and were length measured until they reached gravidity. During early development on fish, body length of attached A. coregoni was negatively correlated with off-host period indicating a mechanism that creates size variance in an attached parasite cohort originally containing equal amounts of resources. However, over time the size differences between parasites became less pronounced and eventually parasites that were kept off-host for longest periods of time reached the length of those individuals that had been allowed to infect a host sooner. A. coregoni thus appears to compensate for delayed growth resulting from an extended host searching period by elevated growth rates, although we show that such accelerated growth incurred a cost, through decreased life-expectancy. PMID- 16255824 TI - Cellular infiltration at skin lesions and draining lymph nodes of sheep infested with adult Hyalomma anatolicum anatolicum ticks. AB - Immunohistochemical analysis of skin and draining lymph nodes of sheep repeatedly infested with the ixodid tick Hyalomma anatolicum anatolicum were studied for different antigen-presenting cells and lymphocyte subpopulations. Infiltration of neutrophils, macrophages and lymphocytes adjacent to the tick bite site were observed. Skin biopsies showed significant increases in dermal infiltration of CD8+ and gammadelta+ T cells at 72 h and 8 days after both primary and secondary infestation. Infiltrations of MHC-II DR/DQ decreased at 72 h after tick infestation, whereas significant increases were recorded for 8-day skin biopsies. CD1+ cellular infiltrations were observed during secondary infestations at the dermis. Decreased ratios of CD4:CD8 T cells and MHC-II:CD1 antigen-presenting cells were observed in both infestations compared to healthy skin biopsies. Ratios of alphabeta:gammadelta T cells increased gradually during infestation compared to uninfested skin. The regional lymph nodes from tick-infested sheep showed an increased CD8+, gammadelta+ T and CD1+ cellular infiltration compared to control lymph nodes. CD4+ T cells were decreased. There were no significant changes in CD45R+ cellular infiltration either at skin lesions or regional lymph nodes. PMID- 16255826 TI - Genetic characterization of parthenogenic Fasciola sp. in Japan on the basis of the sequences of ribosomal and mitochondrial DNA. AB - Accurate identification of aspermic Fasciola forms in Japan remains difficult because of their morphological variations. In order to characterize the forms genetically, nucleotide sequences of ribosomal internal transcribed spacer (ITS1 and ITS2) and mitochondrial cytochrome c oxidase I (COI) and NADH dehydrogenase I (NDI) genes in 34 liver flukes from 16 prefectures in Japan were analysed. Two major forms represented by Fsp 1 and Fsp 2 had sequences identical to or closely resembling those of F. hepatica and F. gigantica, respectively, in all the 4 DNA markers and were mainly distributed in northern and eastern-western parts of Japan, respectively. Fsp 1 and Fsp 2 would have been introduced into Japan with infected cattle of 2 distinct lineages via the Korean Peninsula and spread through limited parts of Japan (northern and eastern-western parts) together with the movement of each cattle lineage. The Japanese form (Fsp 1/2), which showed heterozygosity in ribosomal DNA and Fsp 2 haplotype in mitochondrial DNA, may have originated in interspecific cross hybridization between paternal F. hepatica and maternal F. gigantica. PMID- 16255825 TI - The immunogenicity of Echinococcus granulosus antigen 5 is determined by its post translational modifications. AB - Since its early introduction as a marker for the immunodiagnosis of hydatid disease, antigen 5 (Ag5) has been regarded as one of the more relevant antigens of Echinococcus granulosus, and it is still widely used in different confirmation techniques. In this work we prepared 2 recombinant forms of the antigen, namely, rAg5 (corresponding to the unprocessed polypeptide chain of the antigen) and rAg5 38s (corresponding to its 38 kDa subunit). Their antigenicities were compared to that of the native antigen using a human serum collection. There was a major drop in the reactivity of the sera, particularly against rAg5-38s, which was confirmed by analysis of the cross-reactivity of 2 panels of monoclonal antibodies specific for rAg5-38s and the native antigen. Using the chemically deglycosylated native antigen, we demonstrated that the reduced antigenicity of the recombinants is due to the loss of the sugar determinants, and not to their misfolding. Inhibition experiments using phosphorylcholine confirmed that this moiety also contributes to the reactivity of the antigen, but to a much lesser extent. The presence of immunodominant highly cross-reactive glycan moieties in the Ag5 molecule may involve a parasite evasion mechanism. PMID- 16255827 TI - Effect of phenotypic variation in Xenorhabdus nematophila on its mutualistic relationship with the entomopathogenic nematode Steinernema carpocapsae. AB - The entomopathogenic nematode Steinernema carpocapsae is mutualistically associated with the bacterium Xenorhabdus nematophila. Infective Juveniles (IJs) transport X. nematophila cells that provide them with good conditions to reproduce within the insect. In the laboratory, long term stationary-phase culture conditions sometimes lead X. nematophila's variant 1 cells, which were previously isolated from the worms, to spontaneously and irreversibly change into a new phenotypic variant (variant 2). In this paper, we tested the ability of each phenotypic variant to (i) be transmitted by IJs, (ii) to optimize the worm's fitness within the insect, and (iii) to counteract the effect of closely related antagonistic bacteria previously shown as being able to totally prevent S. carpocapsae's reproduction within the insect. We found that IJs did associate with cells of both phenotypes but that the variant 2 cells were preferentially retained by the nematodes when both variants were present in the insect. Both phenotypic variants led to the same fitness of S. carpocapsae in insects not infected by antagonistic bacteria. In insects infected by antagonistic bacteria, both variants were able to provide protection to S. carpocapsae. Nevertheless, this protection depended on the phenotypic variant and the antagonistic bacteria that were co-injected into the insect. Further analysis conduced in vitro showed that this variability could be partly linked to the sensitivity of each antagonistic bacterium to xenorhabdicin, produced by X. nematophila. PMID- 16255828 TI - Cross-stress tolerance and expression of stress-related proteins in osmotically desiccated entomopathogenic Steinernema feltiae IS-6. AB - Infective juveniles (IJs) of the entomopathogenic nematode (EPN) Steinernema feltiae IS-6 can survive exposure to 24% glycerol solution by entering an osmotically desiccated state. Exposure of osmotically desiccated nematodes to extreme temperature assays (40 degrees C for 10 h and -20 degrees C for 360 h) resulted in gradual reduction in survival, whereas non-desiccated IJs died within a short exposure to the assay conditions. Through SDS-PAGE, a stress-related protein UNC-87 was found in osmotically desiccated IJs exposed to 40 degrees C for 3, 6, and 8 h, whose survival rates were 98.9+/-1.43, 78.5+/-5.87 and 20.9+/ 4.93%, respectively. The protein was not found in IJs following exposure of osmotically desiccated individuals to 40 degrees C for 10 h, in which none of the IJs survived. After exposure to -20 degrees C for 360 h, the survival of osmotically desiccated EPNs with a weak band of UNC-87 was 13.0+/-3.32%. To identify other responsive proteins that are required for osmotic stress, we used 2-dimensional electrophoresis to analyse the proteins in osmotically desiccated EPNs. The results revealed that 10 novel protein spots and 10 up-regulated protein spots in osmotically desiccated IJs were detected by digital image analysis. Mass spectrometry analysis of 7 significant spots indicated that osmotic stress in desiccated IJs was associated with the induction of actin, Proteasome regulatory particle (ATPase-like), GroEL chaperonin, GroES co chaperonin and transposase family member. It seems to show actin, UNC-87 and Proteasome regulatory particle may play distinct roles in specific aspects of organization of macromolecular structures under desiccation stress. GroEL and GroES are members of the Hsp60 family of chaperons. PMID- 16255829 TI - What is the role of p53 during the cyst formation of Trichinella spiralis ? A comparable study between knockout mice and wild type mice. AB - During the cyst formation of Trichinella spiralis, the infected muscle cell undergoes basophilic change and apoptosis, which results in nurse cell formation. This study revealed expression kinetics of some apoptosis genes such as p53 and its closely related genes (tumor suppressor genes p53, p53; mouse double minute 2, MDM2; cyclin-dependent kinase inhibitor p21, p21waf). RT-PCR (reverse transcription polymerase chain reaction) results showed that these genes were temporarily expressed in the infected muscles during the cyst formation period, but not in normal muscles (or very low if any), which suggested the involvement of these apoptosis genes in the nurse cell formation. Cysts and neighbouring muscle cells were separately collected and RT-PCR was performed, which suggested that p53 was expressed in the cysts. An immunocytochemical study showed that p53 was expressed in the nucleoplasm of basophilic cell in the cyst and Trichinella larvae, which suggested involvement of these apoptosis genes in the nurse cell formation. The same p53 expression kinetic study was performed on p53 knockout mice. The knockout mice did not express p53 genes, but expressed the other apoptosis genes in the same kinetics with only minor exceptions, suggesting that the expressions of these genes during the cyst formation were more or less p53 independent. There were no differences in the number and morphology of the cysts between the knockout mice and wild type mice. Thus apoptosis seen during the Trichinella cyst formation can be operated in the presence or absence of p53. PMID- 16255830 TI - Physico-chemical determinants of helminth component community structure in whitefish (Coregonus clupeaformes) from adjacent lakes in Northern Alberta, Canada. AB - Populations of hosts vary extensively in the types and numbers of parasites that the average individual contains. Understanding the factors that lead to this variation is an important goal for parasite ecologists. We characterized patterns of helminth component community structure in whitefish collected from a cluster of 7 lakes located on an isolated plateau in northern Alberta, Canada. Component communities were species rich (5-6 species per lake), high in mean helminth intensity (approximately 80-500 individuals/host), and high in between-lake similarity (50-100%), a pattern consistent with results from studies on whitefish sampled from other localities in Northern Canada and Europe. Multivariate analyses indicated that the structure of the component communities was associated with 2 opposing environmental gradients. One was defined primarily by water colour, the second by phosphorous concentration. Thus, 4 lakes were characterized by a combination of high colour, low productivity, low parasite intensities, and the absence of larval acanthocephalans. Habitat/species associations were less clear as intensities increased, but the 3 remaining lakes tended to have the opposite characteristics. These results provide evidence that variation in helminth component community structure in fish is associated with variation in physicochemical characteristics that are linked to aquatic productivity. PMID- 16255832 TI - Adverse birth outcomes in a malarious area. AB - To determine factors associated with fetal growth, preterm delivery and stillbirth in an area of high malaria transmission in Southern Malawi, a cross sectional study of pregnant women attending and delivering at two study hospitals was undertaken. A total of 243 (17.3%) babies were preterm and 54 (3.7%) stillborn. Intra-uterine growth retardation (IUGR) occurred in 285 (20.3%), of whom 109 (38.2%) were low birthweight and 26 (9.1%) preterm. Factors associated with IUGR were maternal short stature [adjusted odds ratio (AOR) 1.6, 95% confidence interval (CI) 1.0-2.5]; primigravidae (AOR 1.9, 95% CI 1.4-2.7); placental or peripheral malaria at delivery (AOR 1.4, 95% CI 1.0-1.9) and maternal anaemia at recruitment (Hb<8 g/dl) (AOR 1.9, 95% CI 1.3-2.7). Increasing parasite density in the placenta was associated with both IUGR (P=0.008) and prematurity (P=0.02). Factors associated with disproportionate fetal growth were maternal malnutrition [mid-upper arm circumference (MUAC)<23 cm, AOR 1.9, 95% CI 1.0-3.7] and primigravidae (AOR 1.8, 95% CI 1.0-3.1). Preterm delivery and stillbirth were associated with <5 antenatal care visits (AOR 2.2, 95% CI 1.3-3.7 and AOR 3.1, 95% CI 1.4-7.0 respectively) and stillbirth with a positive Venereal Disease Research Laboratory (VDRL) test (AOR 4.7, 95% CI 1.5-14.8). Interventions to reduce poor pregnancy outcomes must reduce the burden of malaria in pregnancy, improve antenatal care and maternal malnutrition. PMID- 16255833 TI - The impact of the Catholic Jubilee in 2000 on infectious diseases. A case-control study of giardiasis, Rome, Italy 2000-2001. AB - Mass gatherings are believed to increase the transmission of infectious diseases although surveillance systems have shown a low impact. The Catholic Jubilee was held in Rome, Italy in 2000. We conducted a case-control study to analyse the risk factors of giardiasis among residents. All diseases reported to the laboratory surveillance system from January 2000 to May 2001 were compared with hospital controls concurrently selected in the same season as cases and frequency matched for age and birth country. Fifty-two cases (44.1%) and 72 controls were enrolled. In the multivariable analysis factors associated with giardiasis among adults were: travelling abroad (OR 24.2, P>0.01), exposure to surface water (OR 4.80, P=0.05), high educational level (OR 3.8, P=0.03). Having a maid from a high prevalence country was independently associated (OR 2.3) although not statistically significant. This is the only exposure that changed during the Jubilee. PMID- 16255834 TI - Survival of a national cohort of hepatitis C virus infected patients, 16 years after exposure. AB - The aim of this study was to describe the natural history of HCV after 16 years of infection, in a cohort of individuals who acquired their infections on a known date in the United Kingdom. A total of 924 HCV-infected transfusion recipients (cases) and 475 anti-HCV negative transfusion recipients (controls) were eligible for inclusion in the study. Survival was compared between cases and controls to see if there was any excess mortality attributable to HCV. The results show that all-cause mortality was not significantly different between cases and controls (hazard ratio 1.17, 95% CI 0.92-1.49, P=0.21). However, the risk of death directly from liver disease was higher in cases than controls (hazard ratio 2.71, 95% CI 1.09-6.75, P=0.03). Nearly 30% of those HCV-infected cases who died directly from liver disease were known to have consumed excess alcohol. PMID- 16255835 TI - Microsatellite analysis of pooled Schistosoma mansoni DNA: an approach for studies of parasite populations. AB - Human parasites are often distributed in metapopulations, which makes random sampling for genetic epidemiology difficult. The typical approach to sampling Schistosoma mansoni involves laboratory passage to obtain individual worms with small sample size and selection bias as a consequence. By contrast, the naturally pooled samples from egg output in stool or urine directly represent the genetic composition of current populations. To test whether pooled samples could be used to estimate population allele frequencies, DNA from individual cloned parasites was pooled and amplified by PCR for 7 microsatellites. By polyacrylamide gel analysis, the relative band intensities of the products from the major alleles in the pooled samples differed by 0-6% from the summed intensities of the individual clones (mean = 2.1%+/-2.1% S.D.). The number of PCR cycles (25-40) did not influence the accuracy of the estimate. Varying the frequency of 1 allele in pooled samples from 32 to 69% likewise did not affect accuracy. Allele frequency estimates from aggregate samples such as eggs will be a better foundation for studies of parasite population dynamics as well as the basis for large-scale association studies of host and parasite characteristics. PMID- 16255836 TI - Associations of pre-trauma attributes and trauma exposure with screening positive for PTSD: analysis of a community-based study of 2,085 young adults. AB - BACKGROUND: While pre-trauma personality and mental health measures are risk factors for post-traumatic stress disorder (PTSD), such information is usually obtained following the trauma and can be influenced by post-trauma distress. We used data collected from a community-based survey of young adults before and after a major natural disaster to examine the extent to which participants' traumatic experiences, demographic and pre-trauma risk factors were associated with their screening positive for PTSD when re-interviewed. METHOD: A representative selection of 2,085 young adults from the Australian Capital Territory and environs, interviewed in 1999 as part of a longitudinal community based survey, were re-interviewed 3-18 months after a major bushfire had occurred in the region. When re-interviewed, they were asked about their experiences of trauma threat, uncontrollable and controllable traumatic experiences and their reaction to the fire. They were also screened for symptoms of fire-related PTSD experienced in the week prior to interview. RESULTS: Four-fifths of participants were exposed to the trauma with around 50% reporting having experienced uncontrollable traumatic events. Reporting PTSD symptoms was associated with being female, having less education, poorer mental health and higher levels of neuroticism prior to the trauma. Particular fire experiences, including being evacuated and feeling very distressed during the disaster, were more strongly associated with PTSD symptoms compared with pre-trauma measures. CONCLUSIONS: While demographic and pre-trauma mental health increased the likelihood of reporting PTSD symptoms, exposure to trauma threat and reaction to the trauma made greater contributions in explaining such symptoms as a result of this disaster. PMID- 16255837 TI - Cutaneous glucocorticoid receptor sensitivity and pro-inflammatory cytokine levels in antidepressant-resistant depression. AB - BACKGROUND: There is evidence to indicate that peripheral glucocorticoid receptor (GR) function is reduced in major depression, and a possible molecular explanation for this is the impact of raised pro-inflammatory cytokines. The topical steroid vasoconstriction assay provides a convenient probe of peripheral GR function. The present study sought to assess the sensitivity of peripheral GRs in antidepressant-resistant major depressives and investigate the association between GR sensitivity and circulating plasma cytokines. METHOD: Nineteen antidepressant-resistant depressives together with age- and sex-matched healthy controls underwent the steroid vasoconstriction assay using three commercial preparations of corticosteroids containing clobetasol propionate 0.05%, betamethasone valerate 0.1%, and clobetasone butyrate 0.05%, corresponding to very potent, potent, and moderately potent steroid creams respectively. The pro inflammatory cytokines, tumour necrosis factor-alpha (TNF-alpha) and interleukin 6 (IL-6) were measured using enzyme-linked immunosorbent assays. The severity of the depressive episode was assessed using the Hamilton Depression Scale (HAMD). RESULTS: Depressed subjects had a significantly reduced vasoconstriction response across all three strengths of steroid. They also had significantly higher concentrations of TNF-alpha and IL-6. There was a significant inverse correlation between TNF-alpha concentration and vasoconstriction response and also between the HAMD score and vasoconstriction response. CONCLUSIONS: These findings suggest that cutaneous GR function is abnormal in antidepressant-resistant depression, that circulating TNF-alpha may play a significant role in this abnormality and that the efficacy of topical steroids in antidepressant-resistant depressives is reduced. PMID- 16255838 TI - Effectiveness of the Coping With Depression (CWD) course for older adults provided by the community-based mental health care system in the Netherlands: a randomized controlled field trial. AB - BACKGROUND: The Dutch version of the Coping With Depression (CWD) course for older adults has been implemented in the prevention arm of the community-based mental health care system in the Netherlands. The study group included older adults with subclinical depression as well as those with a major depressive disorder; all were enrolled into the course by mental health care professionals. The effectiveness (immediate and long-term) of the course for this heterogeneous population was studied in an effectiveness trial. METHOD: Participants were self referred, responding to media announcements. A total of 119 participants aged 55 85 years (69% female), with subclinical depression and major depression, were randomized to either the CWD course (N = 61) or the waiting list (N = 58). RESULTS: Nine participants dropped out of the course. According to a diagnostic interview based on the DSM-IV, 39% had a major depressive disorder (MDD), 69% had had a previous MDD, and 45% had an anxiety disorder. Older adults in the intervention group showed a significant decrease in depression symptoms. Gains were maintained over 14 months. In the intervention condition 83% had a pre treatment score > or = 16 on the Center for Epidemiologic Studies Depression Scale (CES-D); at post-treatment 62% still scored > or = 16. CONCLUSIONS: The course was beneficial for participants with mild or severe depression, and treatment acceptability was high. It should be fitted into a stepped-care protocol that varies intervention intensity according to clinical needs, using the post-treatment level of functioning as an indication for the next step. PMID- 16255839 TI - Use of aripiprazole for psychosis and agitation in dementia. AB - We report the case of an elderly woman with Alzheimer's disease, risk factors for vascular dementia, and atrium-ventricular blockade, who presented with severe agitation and psychosis. She was treated with aripiprazole and assessed with the Neuropsychiatric Inventory (NPI) over the course of 14 weeks. NPI scores showed a marked decrease in psychosis and agitation at week 4, and complete recovery at week 14, except for depression. PMID- 16255840 TI - The impact of residential respite care on the behavior of older people. AB - BACKGROUND: The aim of this study was to examine the impact of residential respite care on disruptive behavior displayed by older people, particularly those with dementia. METHODS: A quasi-experimental, repeated-measures, single-group design was used. The participants were a consecutive series of 100 older people with a mean age of 81.8 years (range 66-96 years) who had been booked for a respite admission to one of several residential aged care facilities in a provincial Australian city. A diagnosis of dementia was reported for 29% of the sample. Disruptive behaviors were rated before and after the period of respite by home caregivers (N = 100) and during the period of respite by nurses (N = 25) using the Dementia Behavior Disturbance Scale (DBDS). RESULTS: Age, male gender and the presence of dementia were all significantly related to the frequency of reported disruptive behaviors. Residential respite care was associated with a significant reduction in the frequency of reported disruptive behaviors in older people (Wald chi2 = 28.28, p < 0.0001). However, this improvement in behavior did not persist into the post-respite period. The deteriorating behavioral trajectory that was evident prior to respite care continued following the period of respite care. CONCLUSIONS: Residential respite care was associated with a temporary diminution in the frequency of reported disruptive behaviors in older people. This finding should be reassuring both for family carers considering placing a relative in residential respite care and for health workers considering whether to recommend such a course of action. PMID- 16255841 TI - The Short Cognitive Performance Test (SKT): a preliminary study of its psychometric properties in Brazil. AB - BACKGROUND: Most instruments designed to detect dementia can lack appropriate sensitivity in the early stages of Alzheimer's disease (AD), and are subject to educational bias. The Short Cognitive Performance Test (Syndrom-Kurztest, SKT) is considered a suitable instrument to measure cognitive decline as it assesses memory, attention, and related cognitive functions, taking into account the speed of information processing. OBJECTIVES: The aim of this study was to examine the psychometric characteristics of the SKT as a dementia screening instrument in a Brazilian population sample, as compared to the Mini-mental State Examination (MMSE) and the Clock-Drawing Test (CDT). The effect of educational level on performance in the three screening tests was also verified. METHODS: Fifty-one elderly subjects were assessed. Consensus diagnoses were established by an expert multidisciplinary team, considering clinical, neuropsychological and neuroimaging data. Subjects were further classified into those with (1) mild and moderate AD, (2) non-Alzheimer's dementia, (3) mild cognitive impairment, and (4) controls, according to National Institute for Communicative Disorders and Stroke- Alzheimer's Disease and Related Disorders Association (NINCDS-ADRDA) criteria. RESULTS: Statistical analyses revealed high internal consistency for the SKT (Cronbach's alpha = 0.80) and significant correlations between the total score and the SKT subscores separately (p < 0.01). Comparison of the three tests revealed strong correlations between the SKT and the MMSE (r = -0.66, p < 0.0001) and between the SKT and the CDT (r = -0.57, p < 0.0001). The SKT, MMSE and CDT scores were correlated with education. CONCLUSIONS: The Brazilian version of the SKT maintains its original psychometric properties and displays significant correlation with previously validated screening tools for dementia. Like other dementia screening tests, the SKT is subject to educational bias. PMID- 16255842 TI - Immunological effects of refolded human soluble BAFF synthesized in Escherichia coli on murine B lymphocytes in vitro and in vivo. AB - The B cell activating factor of the TNF family (BAFF, also known as BLyS, TALL-1, THANK, and zTNF4) is an important survival factor for B lymphocytes. Our previous study has demonstrated that the final purified material of human soluble BAFF (refolded hsBAFF) synthesized in Escherichia coli is biologically active in a validated induced human B lymphocyte proliferation bioassay. In this study, the administration of refolded hsBAFF to isolated mouse B lymphocytes and mice was carried out to study the immunological effects of hsBAFF on in vitro and in vivo B lymphocytes. The results showed that splenic B lymphocyte proliferation significantly increased after hsBAFF administration (in vitro 1, 2, 3, 5 microg/ml and in vivo 0.01, 0.5, 1.0 mg/kg body mass). An oppositely elevated immune response of B lymphocyte to LPS stimulation after hsBAFF administration (1, 2.5, 5 microg/ml) and a significantly elevated change after treatment with hsBAFF and costimulation with anti-IgM (2.5 microg/ml) was observed in vitro, respectively. A similar change existed also in hsBAFF-treated mice on the 8th postexperiment day, but the value with anti-IgM alone didn't increase compared to normal control in vitro. We found that the treatment of mice with hsBAFF resulted in a developmental maturation of T1 B lymphocytes to T2 and mature B lymphocytes by detecting distributions of splenic CD21(lo) with CD45R/B220(+) and CD21(hi) with CD45R/B220(+) subsets. These results suggest that the refolded hsBAFF synthesized in Escherichia coli may enhance immune responses in the body by regulating the proliferation, differentiation, and immune response of B lymphocytes. PMID- 16255843 TI - Effect of hypothyroidism on intestinal zinc absorption and renal zinc disposal in five-sixths nephrectomized rats. AB - Both hypothyroid (Hypo) and hypozincemia are commonly observed in patients and animals with chronic renal failure (CRF). In CRF whether the hypothyroid plays a role in the pathogenesis of hypozincemia is unclear. This study is designed to investigate the effects of hypothyroid on intestinal zinc absorption and urinary zinc excretion in 5/6 nephrectomized (Nx) rats, because plasma zinc balance is attained through a controlled rate of intestinal uptake as well as renal reabsorption. Intestinal zinc absorption was carried out in jejunum and ileum segments by an in vivo perfusion technique and the renal zinc disposal was evaluated by a conventional method using a standard formula to calculate the zinc tubular reabsorption and the excretion of urinary zinc in 5/6 Nx rats with hypothyroidism. The Hypo-NxT rats showed a significant decrease in the rate of intestinal zinc absorption and in the response of plasma zinc levels during intestinal zinc perfusion compared with Eu-NxT rats. They also had significantly lower levels of mucosal zinc and MT as well as lower content of liver zinc than Eu-NxT rats after intestinal zinc perfusion for 80 min. Hypo-NxT rats showed low plasma zinc levels, but had a similar output of pancreaticobiliary zinc and excretion of 24-h urine zinc compared with the Eu-NxT rats. When 2% alcohol intestinal perfusion was used to produce water diuresis, the Hypo-NxT rats presented a higher excretion of urinary zinc than the Eu-NxT rats did, especially during 2% alcohol intestinal zinc perfusion. In the Hypo-NxT rats, the lower plasma zinc levels may thus result from the hypothyroid because it reduces intestinal zinc absorption. Increasing the urine flow rate may aggravate the reduction of plasma zinc level in Hypo-NxT rats because of the increased excretion of urinary zinc. PMID- 16255845 TI - [Pay much attention to prevention of adverse drug reactions]. PMID- 16255844 TI - Variable Unstressed Volume Keeps Normal Distributions of Canine Left Ventricular Contractility and Total Mechanical Energy under Atrial Fibrillation. AB - We have reported that the contractility index (E(max)) and the total mechanical energy (PVA) of arrhythmic beats of the left ventricle (LV) distribute normally in canine hearts under electrically induced atrial fibrillation (AF). Here, E(max) is the ventricular elastance as the slope of the end-systolic (ES) pressure-volume (P-V) relation (ESPVR), and PVA is the systolic P-V area as the sum of the external mechanical work within the P-V loop and the elastic potential energy under the ESPVR. To obtain E(max) and PVA, we had to assume the systolic unstressed volume (V(o)) as the V-axis intercept of the ESPVR to be constant despite the varying E(max), since there was no method to obtain V(o) directly in each arrhythmic beat. However, we know that in regular stable beats V(o) decreases by approximately 7 ml/100 g LV with approximately 100 times the increases in E(max) from ~0.2 mmHg/(ml/100 g LV) of almost arresting weak beats to approximately 20 mmHg/(ml/100 g LV) of strong beats with a highly enhanced contractility. In the present study, we investigated whether E(max) and PVA under AF could still distribute normally, despite such E(max)-dependent V(o) changes. The present analyses showed that the E(max) changes were only approximately 3 times at most from the weakest to the strongest arrhythmic beat under AF. These changes were not large enough to affect V(o) enough to distort the frequency distributions of E(max) and PVA from normality. We conclude that one could practically ignore the slight E(max) and PVA changes with the Emax-dependent V(o) changes under AF. PMID- 16255846 TI - [Influence of methylphenidate on growth of school age children with attention deficit hyperactivity disorder]. AB - OBJECTIVE: To determine whether long-term treatment of attention deficit hyperactivity disorder (ADHD) with methylphenidate influences the growth in height and weight of children. METHODS: Analyses were performed on 146 school age children (126 boys) diagnosed as ADHD and treated with methylphenidate [0.27-0.64 mg/(kg.day)] for methylphenidate group and 29 children with ADHD who did not receive any medication for ADHD (controls). These children were followed-up for 2 4 years. Changes in height and weight after long-term treatment with methylphenidate were recorded and the factors affecting growth of height, weight, and height velocity were analyzed. RESULTS: The change of difference between patients' height and mean height in methylphenidate group and controls was (-1.86 +/- 0.82) cm (paired t test, t = 27.335, P < 0.001) and (-0.26 +/- 0.51) cm (P < 0.05), respectively; the change of height standard deviation score (SDS) in methylphenidate group and controls was -0.14 +/- 0.23 SD (paired t test, t = 7.326, P < 0.001) and +0.05 +/- 0.10 SD (P < 0.05), respectively. When the height change and height SDS change in methylphenidate group and controls were compared by using independent-samples T-test, the t value was -10.078 and -4.262 respectively, P for both was < 0.001. Both of bivariate correlation analysis and stepwise multiple-regression analysis indicated that the duration of treatment contributed significantly to the variance in change of height (P < 0.001); but age, sex, DSM-IV type, NJ22 degree and dose of methylphenidate did not contribute significantly to the variance of height. The mean height velocity from 1st to 4th year was 4.28 cm/year, 4.90 cm/year, 4.98 cm/year and 4.95 cm/year, respectively. With Friedman test, Chi-square = 253.673, P < 0.001. The change of difference of patients' weight to weight for height after methylphenidate was (-0.14 +/- 1.25) kg (paired t test, t = 1.326, P > 0.05). CONCLUSION: Small but significant deceleration of height velocity is the identified long-term side effect of methylphenidate, the magnitude of height deficit is related to duration of treatment. The height velocity was significantly attenuated in the first year. Methylphenidate had no significant influence on weight. PMID- 16255848 TI - [Acute benign myositis--a case report]. PMID- 16255847 TI - [Effect of carbamazepine and valproate on bone metabolism in children with epilepsy]. AB - OBJECTIVE: To assess bone health in epileptic children who have been treated with carbamazepine (CBZ) or valproate (VPA) by using quantitative ultrasound (QUS) and determining the biochemical indices of bone metabolism, and to provide a proposal to improve quality of life of epileptic children. METHODS: Ninety-two epileptic children who had been treated with CBZ or VPA for more than two years were evaluated for bone mineral density (BMD) at the mid-shaft tibia and the distal third of the radius. Biochemical indices of bone metabolism including urine deoxypyridinoline (DPD) and serum osteocalcin (OC), and daily calcium intake were also evaluated. Thirty-five age-matched healthy children were used as controls. Reduced BMD was defined as speed of sound (SOS) Z scores of the mid-shaft tibia and (or) the distal third of the radius less than -0.7. RESULTS: BMD was reduced in epileptic children significantly when compared to the controls (P < 0.05). In addition, a negative correlation was found between the duration of anti-epileptic drugs (AEDs) use and BMD (r(s) = -0.21 - -0.31, P < 0.05), the lowest BMD was observed in those who had been treated for the longest time. The serum values of OC in epileptic children were significantly reduced relative to the controls (P < 0.01), children who took VPA had the lowest value of OC. However, the urine values of DPD showed no significant difference between epileptic and healthy children (P > 0.05); children who took CBZ had the highest value of DPD. Thirty two epileptic children (35%) and five (14%) sex- and age-matched healthy children had reduced BMD, significant difference was found between them (P < 0.05). Moreover, epileptic children with reduced BMD seemed to have higher body mass index (BMI) (P < 0.05), take less daily calcium intake (P < 0.01), and had longer duration of AEDs (P < 0.01). The two risk factors of having reduced BMD in epileptic children were those who had been treated with AEDs for more than five years and higher BMI, while the protective factor was daily calcium intake. CONCLUSIONS: Long-term use of CBZ or VPA is associated with bone metabolism abnormalities, which include reduced BMD and decreased bone turnover (mainly decreased bone formation). Long-term anti-epileptic therapy is an important factor for impaired bone health in epileptic children, and that low calcium intake and high BMI could be two aggravating factors. QUS is a useful method to evaluate BMD of epileptic children who are on long-term anti-epileptic therapy, and to recognize the status of bone health, in helping to promote bone health and improve quality of life in epileptic children by the use of calcium and vitamin D supplementation. PMID- 16255849 TI - [Expression of multidrug resistance gene and topiramate affect expression of multidrug resistance gene in the hippocampus of spontaneous epileptic rats]. AB - OBJECTIVE: Refractory temporal lobe epilepsy (TCE) shows a unique type of hippocampal damage, referred to as hippocampal sclerosis. The mechanisms underlying drug-refractoriness in TCE are poorly understood, which may be connected with pharmacoresistance to antiepileptic drugs (AEDs). Some studies show that expression of the multidrug resistance gene (mdr1a and mdr1b) and p glycoprotein encoded by mdr1a and mdr1b are high in the brain, especially in the hippocampus, and the expression may lead to reduction of AEDs concentration in the brain. But most of these studies focused on acute epileptic activity shortly after status epilepticus (SE), spontaneous seizures are seldom studied. The authors used a rat model of kainic acid induced spontaneous seizures to investigate expression of mdr1a and mdr1b mRNA, and explore whether topiramate (TPM) affects expression of mdr1a and mdr1b in the hippocampus. METHODS: Seizures were induced by intraperitoneal injection of 10 mg/kg kainic acid at postnatal day 28. Control rats were injected with sodium chloride. All rats were divided into 4 groups 1 week after spontaneous seizures developed: status epilepticus complicated with spontaneous seizures (SE, n = 8) group, status epilepticus complicated with spontaneous seizures treated with TPM (SE + TPM, n = 9) group, spontaneous seizures without status epilepticus (N-SE, n = 7) group, spontaneous seizures without status epilepticus treated with TPM (N-SE + TPM, n = 8) group, control (n = 7) group and control treated with TPM (control + TPM, n = 7) group. The treated rats were given therapeutic dose of TPM (25 mg/kg). All the rats were killed on the 42nd day of administration. The mdr1a and mdr1b mRNAs in the hippocampus were measured by RT-PCR. RESULTS: Expression of mdr1a and mdr1b mRNA in the hippocampus increased significantly in the SE + TPM group, SE group and N SE + TPM group compared with control group (P < 0.001 or < 0.05). The mRNA in SE + TPM group increased significantly compared with the SE group, too (P < 0.01). The mdr1a and mdr1b mRNA expression in the hippocampus in control + TPM and N-SE groups did not change. CONCLUSION: Frequent seizures, especially status epilepticus resulted in overexpression of mdr1a and mdr1b mRNAs in the hippocampus. The drug-refractoriness mechanism in TCE may be related to overexpression of mdr1a and mdr1b mRNAs. TPM could enhance the expression of mdr1a and mdr1b mRNAs in the hippocampus. Seizure activity and TPM are likely to be the main determinant in enhancing mdr1a and mdr1b mRNA expression in epilepsy. PMID- 16255850 TI - [Bibliometric analysis of Chinese Journal of Pediatrics]. AB - OBJECTIVE: To evaluate the quality of articles and the popularity of Chinese Journal of Pediatrics. METHODS: With the database of Chinese Science & Technology Journal Citation Reports 2000-2003, the six indicators, including impact factor (IF), total cited numbers, immediate index, ratio of cited numbers by other journals to total cited numbers, number of citing journals, and cited half-life were evaluated and compared with other Chinese periodicals in pediatrics and related subjects. The cited numbers, distribution of cited areas, and cited institutions of published articles in Chinese Journal of Pediatrics 2000-2003 were analyzed. RESULTS: From 2000 to 2003, the impact factor of the Journal was 0.937, 1.025, 1.052, 1.252, total cited number was 1378, 1468, 1863 and 2396. Immediate index, citing journals, and cited half-life rose steadily, and the ratio of cited numbers by other journals to total cited numbers remained above 0.9. The average cited number of articles was 3.3. The cited authors were from 26 provinces or cities. CONCLUSION: The Chinese Journal of Pediatrics ranked at the top of Chinese pediatric journals with its highest IF and total cited numbers, which indicates that the journal has explicit editorial aims, stable continuity, and the increased influence in Chinese pediatric research. The stable growth of immediate index, number of citing journals and cited half-life show that the Journal has a high rate of reflection, the articles published in the Journal were paid more attention and have influenced researchers for longer periods and widely. The higher rate of citation by others and lower self-cited rate indicate that the Journal has been rigorous and realistic in editorial practice. The Journal is demonstrated to be one of the main medical core periodicals and one of the most important information sources in the field of pediatrics in China. PMID- 16255851 TI - [Role of genetic factors in occurrence of neonatal jaundice in Guangxi region]. AB - OBJECTIVE: To investigate the possible role of the UGT1A1 G71R or the OATP2 A388G genes mutation with coexisting G-6-PD deficiency in occurrence of neonatal hyperbilirubinemia. METHODS: Totally 109 umbilical cord blood samples were collected for the screening of G-6-PD activity by nitroblue tetrazolium (NBT) test and identification of G71R gene type by polymerase chain reaction combined with allele-specific oligonucleotide assay (ASO-PCR). At the same time, for 101 of the 109 cord blood samples A388G gene types were tested by restriction fragment length polymorphism (RFLP). According to G-6-PD activity and G71R or A388G genotype, the neonates were allocated into different groups. The authors compared the incidence rate of hyperbilirubinemia among different groups. Ten samples were sequenced to validate the results. RESULTS: Among the 109 umbilical cord blood specimens, the allele frequency of G71R was 22.03% in G-6-PD deficient group, 28.0% in G-6-PD normal group. The incidence rate of neonatal hyperbilirubinemia for those neonates who were G-6-PD deficient with coexisting homozygous or heterozygous variant of the G71R gene was significantly higher than that of neonates who were G-6-PD normal and had wild type G71R gene type, chi(2) = 10.45, P = 0.0012. The odds ratio (OR) of the former was 18.00 (95% CI: 2.12, 152.9). Among the 101 neonates, the allele frequency of A388G was 20.0% in G-6-PD deficient group and 18.5% in G-6-PD normal group. The incidence rate of neonatal hyperbilirubinemia for those neonates who were G-6-PD deficient with coexisting homozygous or heterozygous variant of the A388G gene was significantly higher than that of the neonates who were G-6-PD normal and had wild type A388G gene type, chi(2) = 10.39, P = 0.0013. The OR of the former was 11.8 (95% CI: 2.15, 56.48). CONCLUSION: G-6-PD deficiency coexists with G71R or A388G mutation in some individuals in Guangxi region. UGT1A1 G71R gene mutation combined with G-6 PD deficiency or A388G gene mutation combined with G-6-PD deficiency may play a coordinative role in the development of neonatal hyperbilirubinemia. PMID- 16255852 TI - [Effects of selective head cooling on cerebral blood flow and cerebral metabolic rate in newborn piglets]. AB - OBJECTIVE: To study the change in regulation of cerebral blood flow (CBF) and cerebral metabolism rate (CMR) during selective head cooling (SHC) in newborn piglets. METHODS: Sixteen newborn piglets, aged 5 approximately 7 days, were randomly assigned to one of the following three groups: SHC in normal piglets (n = 4), SHC after hypoxia-ischemia HI (n = 6) and normal temperature after HI (n = 6). Nasopharyngeal temperature was respectively reduced to 35 degrees C and then 32 degrees C in SHC group. HI was induced by using temporary occlusion of the bilateral carotid arteries and mechanical ventilation with low concentration of oxygen (6%) for 30 minutes. CBF was measured with color microspheres method. Mixed venous blood and arterial sample were collected at the same time from superior sagittal sinus and forelimb artery respectively for blood gas analysis, blood glucose and lactate measurement. Cerebral oxygenation metabolism rate (CMRO(2)), cerebral glucose metabolism rate (CMR(Glu)) and cerebral lactate production (CLP) were calculated. RESULTS: In normal piglets, CBF, CMRO(2) and CMR(Glu) were significantly decreased at 35 degrees C and 32 degrees C respectively, while CLP did not change. When nasopharyngeal temperature decreased to 35 degrees C, The ratios of CBF/CMRO(2), CBF/CMR(Glu) and CBF/CLP did not significantly change, coupling of CBF and CMR was maintained. While nasopharyngeal temperature decreased to 32 degrees C, the ratios of CBF/CMR(Glu) and CBF/CLP significantly decreased. Maintained newborn piglets normal temperature after HI, the CBF and CMRO(2) were significantly reduced, while CMR(Glu) and CLP were markedly increased. The ratio of CBF/CMRO(2) increased, CBF/CMR(Glu) and CBF/CLP decreased. Uncoupling of CBF and CMR was observed. During SHC after HI, the same decrease was observed in CBF and CMR(Glu) and CLP was markedly reduced. The ratio of CBF/CMRO(2), CBF/CMR(Glu) and CBF/CLP were restored at 35 degrees C and 32 degrees C as compared to the same time point in normal temperature, respectively. CONCLUSION: CBF and CMR decreased in normal piglets during SHC and coupling of CBF and CMR was maintained. After HI, CBF was reduced and the disturbance of cerebral oxygenation metabolism occurred. CBF and CMR was uncoupled. SHC after HI can improve CMR and correct uncoupling of CBF and CMR. PMID- 16255853 TI - [Neonatal lupus erythematosus in a case]. PMID- 16255854 TI - [Expression of connective tissue growth factor in progressive muscular dystrophy]. AB - OBJECTIVE: Progressive muscular dystrophy (PMD) is characterized by muscle fiber necrosis, regeneration, and endomysial fibrosis. Although absence of dystrophin and subsarcolemmic protein has been known as the cause of muscle fiber degeneration, pathogenesis of interstitial fibrosis is still unknown. The aim of this study was to investigate the role of connective tissue growth factor (CTGF) in PMD and its relationship with muscular fibrosis. METHODS: Immunological localization of CTGF was examined in frozen muscle specimens obtained via biopsy from 8 patients with Duchenne muscular dystrophy (DMD), 2 patients with Becker muscular dystrophy (BMD), 6 patients with congenital muscular dystrophy (CMD) and 6 cases with normal muscle by immunohistochemistry, double immunofluorescence and Western blot analysis. RESULTS: The results of immunohistochemistry and double immunofluorescence showed that CTGF was positive only in vessels of normal muscle. Both immunohistochemistry and Western blot analysis showed that CTGF expression was distinctly increased in dystrophy muscles of PMD than that in normal muscles. In dystrophy muscle, marked immunostaining of CTGF was not only observed in vascular walls, but also strongly expressed in the cytoplasm and nuclei of regenerating muscle fibers, and also immunolocalized in the muscle fiber sarcolemma of non-regenerating fibers. Double labeling with antibodies against CTGF and CD68 demonstrated that CTGF was expressed in some macrophages and some macrophage infiltrated necrotic fibers. CTGF was strongly expressed in endomysial and perimysial connective tissues of dystrophy muscles of patients with DMD, CMD and FCMD. Double immunolabeling revealed that most activated fibroblasts in perimysium and endomysium were positive for CTGF, but not all of connective tissues were co-localized with CTGF. Older cases with FCMD showed poor or no expression of CTGF in advanced fibrosis. CONCLUSION: CTGF may play a role in the pathogenetic process of muscular dystrophy, and CTGF may be important for muscle repair and fibrosis. PMID- 16255855 TI - [Diagnostic value of echocardiography for cardiac tumors in infancy and childhood]. AB - OBJECTIVE: The present study was designed to evaluate the clinical manifestations, surgical findings, pathologic types and treatment of cardiac tumor and to analyze the echocardiographic characteristics of the cases. METHODS: Records of 19 patients with cardiac tumors confirmed by operations and pathology at Fuwai Cardiovascular Hospital in Beijing, China between Jan, 1983 and Dec, 2003 were reviewed. Clinical and echocardiographic data of all patients were analyzed. RESULTS: The median age of patients was 7 +/- 5 years, ranging from 5 months to 14 years. There were 8 male and 11 female cases. The surgical findings revealed that 57.9% (11 cases) of cardiac tumors were found in left heart, 36.8% (7 cases) in right heart, 5.3% (1 case) in two ventricles. The pathological study revealed that 17 cases (89.5%) were diagnosed as primary cardiac benign tumors including myxoma in 10 cases (52.6%), rhabdomyoma in 4 cases (21.1%), fibroma in 2 cases (10.5%) and lipoma in 1 case (5.3%). Two cases were diagnosed (10.5%) as cardiac malignant tumors including a primary cardiac rhabdomyosarcoma and a metastatic epithelioid sarcoma. By using echocardiography 11 cases were diagnosed as myxomas and rhabdomyoma (11/19, 57.9%), 8 cases were diagnosed as space occupying lesions of the heart or myxoma (8/19, 42.1%). CONCLUSIONS: Myxomas are the most common heart tumors seen in infancy and childhood, followed in frequency by rhabdomyomas, fibromas and lipomas. Surgery is recommended for patients with refractory and severe clinical symptoms. PMID- 16255856 TI - [Early intervention on atherosclerosis by fluvastatin and lectin-like oxidized low-density lipoprotein receptor-1 expression in atherosclerotic arteries in immature rabbits]. AB - OBJECTIVE: The present study was designed to investigate the preventive and therapeutic effect of 3-hydroxy-3-methylglutanyl coenzyme A (HMG-CoA) reductase inhibitor fluvastatin on the development of atherosclerosis (AS) in immature rabbits and its possible mechanism by detecting the expression level of lectin like oxidized low-density lipoprotein receptor-1 (LOX-1) in the abdominal aorta. METHODS: A model of hypercholesterolemia (HC) was established by high-cholesterol diet and 24 immature rabbits were divided randomly and equally into control group, HC-diet group and fluvastatin group. At the beginning of the study and after 12 weeks, the body height (BH) and body weight (BW) of the rabbits were measured and their body mass index (BMI) was calculated. At the end of 12 weeks, serum total cholesterol (TC) and low-density lipoprotein (LDL) levels were examined. The intima-medial thickness of the abdominal aorta (aIMT) was measured by using non-invasive high-resolution (14 MHz) B-mode ultrasound imaging. Histological changes in abdominal arteries were studied by H&E-staining and histomorphometric analysis. The gene expression of LOX-1 in abdominal aorta was evaluated by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) and its protein expression was examined by immunohistochemistry. RESULTS: High cholesterol diet induced hypercholesterolemia and early AS in immature rabbits. In HC-diet group serum TC and LDL levels in rabbits elevated. B mode echocardiography showed that aIMT was thickened and pathomorphology indicated that extensive aortic intima (I) and intima and media (I + M) became thickened and the ratio of the area of intima to media (S(I)/S(M)) was increased. Aortic intimal proliferation in HC-diet group was associated with a marked increase in LOX-1 expression (protein and mRNA) in endothelium and neointima of the abdominal aorta. Treatment with fluvastatin at a dosage of 10 mg/(kg.d) deduced serum lipid, attenuated artery intimal proliferation and markedly decreased the enhanced LOX-1 expression level in endothelium and neointima in immature rabbits. There were no significant differences of BH, BW or BMI among the three groups. CONCLUSIONS: These findings suggested that early treatment with fluvastatin not only induced a significant regression of arterial lesions of HC and early AS in immature rabbits, but also had a crucial endothelial protective effect by down-regulating LOX-1 expression level in atherosclerotic arteries in early AS. PMID- 16255857 TI - [Risk factors for arrhythmia early after transcatheter closure of perimembranous ventricular septal defects]. AB - OBJECTIVE: Perimembranous ventricular septal defects (PMVSDs) is the most common type of congenital ventricular septal defects (VSD), which accounts for 70% approximately 80% of VSD. The structure of PMVSDs is very complex, it is close to tricuspid valve, mitral valve and aortic valve. The atrioventricular (AV) node is located in the posterior upper membranous ventricular septum and branches into left and right bundle in the posterior lower margin. This increases the risk of transcatheter closure of PMVSDs. Arrhythmias is the common complication after transcatheter closure of PMVSDs. The present study aimed to identify the risk factors resulting in arrhythmias after transcatheter closure of PMVSDs in patients under 18 years of age to decrease the incidence of arrhythmias after the interventional catheterization. METHODS: A retrospective analysis was performed on the patients treated with transcatheter intervention from June 2002 to June 2004. Transcatheter closure of PMVSDs with Amplatzer membranous septal occluder and a domestic product was performed in 89 cases after obtaining consent themselves and/or their guardian or parents, 47 cases were males and 42 females. The age of the cases ranged from 3 to 18 years (mean 8.2 years) and the body weight ranged from 13 to 55 kg (mean 26.7 kg). They were all diagnosed as having PMVSDs with trans-thoracic echocardiography (TTE) before the interventional catheterization, the electrocardiographic (ECG) and chest X-ray (CXR) findings were recorded. A simultaneous care ECG and TTE were performed during operation in order to identify the effect of the transcatheter closure, the heart structure and functional changes and whether or not arrhythmias occurred, respectively. In 80 cases AGA Amplatzer membranous septal occluder was used and in 9 cases a domestic product was used. Follow-up was performed based on the echocardiography and ECG. RESULTS: The devices were successfully implanted in 89 cases; 11 cases (12%) developed various block of heart conduction within 5 days, which included first degree AV block in 1 patient, third degree AV block in 1, left anterior bundle branch block in 5, partial right bundle branch block in 4, complete right bundle branch block in 3, and 3 patients had two kinds of heart block. Eight patients were treated with corticosteroids, 6 of them recovered within 14 days, 1 patient within 1 month and in 1 case the problem shifted from first degree block and left anterior bundle branch block to left anterior bundle branch block 5 days later and that persisted for 6 months. It was found that the distance from upper margin of defects to the aortic valve < 3 mm, the diameter of ventricular septal defect > or = 8 mm, the diameter of device > or = 10 mm, blood pH < 7.35 and arteriovenous track building time after the success of the Seldinger technique > or = 60 min were independent predictors of post-closure arrhythmias. CONCLUSION: Arrhythmias remain the severe early complications after interventional catheterization for PMVSDs in patients under 18 years of age. Shortening of operation time, prevention of acidosis and strict selection of indications may be the most effective measures to prevent arrhythmias after transcatheter closure of PMVSDs in patients under 18 years of age. PMID- 16255858 TI - [Spatial expression of cardiac alpha-actin during heart development in embryos]. AB - OBJECTIVE: Conotruncal defects (CTD) is a common type of cyanotic congenital heart defects. It was shown that CTD might be produced by ablation of cardiac neural crest during early stage embryos in experimental studies. There were many kinds of genes involved and cardiac alpha-actin (CAA) was one of them. The purpose of this study was to investigate the spatial expression of CAA gene during embryonic heart development, and to explore its role in the pathogenesis of congenital heart defects. METHODS: Experimental chicken embryos were subjected to cardiac neural crest ablation by using electric stimulation at 36-40 hr incubation stage. The embryos and organs of experimental and normal control groups during different developing periods were taken out, RNA was extracted by Trizol, and then reverse transcription was done. The dynamic changes of CAA mRNA expression at different development stages of embryos were assayed by fluorescent real-time PCR. The embryos and tissues at different stages were taken out, and paraffin sections were made. The localizations of CAA antibody expression in the developmental embryos were detected by immunohistochemical analysis (peroxidase DAB coloration). RESULTS: (1) The expression of CAA gene was detected at early embryo development, and increased subsequently to a stable level. Its expression was mainly limited to heart site, and could be increased along with the maturation of the cardiac muscle. There was no expression or little trace expression in liver, brain, and stomach. (2) There was a significantly low level of CAA gene expression on days 2-7 of chicken embryos whose cardiac neural crest were ablated in comparison with normal controls (P = 0.013). The level of CAA gene expression was also down-regulated on days 7, 9 and 15 of embryonic cardiac tissues (P = 0.029). CONCLUSION: CAA gene is closely associated with heart development, its expression was adjusted by cardiac neural crest, and its dysfunction may be an important factor leading to congenital heart defects. PMID- 16255859 TI - [Prognoses of food allergy in infancy]. AB - OBJECTIVE: Food allergy as one of social health problems has gained more attention of people. However, few reports on prognosis of food allergy, the relation between infant food allergy and other allergic diseases, and factors affecting prognoses of infant food allergy in China. The present study was designed to investigate the evolution of infancy food allergy and to explore predicting factors of the tolerance to these foods and factors of other allergic diseases to provide a clue for managing children with food allergy reasonably and to improve their prognoses. METHODS: Totally 119 children with food allergy during infancy, who were diagnosed in the Department of Primary Child Care, Children's Hospital, Chongqing University of Medical Sciences from January 2000 to June 2003, were studied retrospectively by analyzing the follow-up data before March 2004. The occurrence of food tolerance and other allergic diseases were counted. Kaplan-Meier methods were used to calculate the cumulative tolerance probability. And the predicting factors of persistent food allergy and influential factors of other allergic diseases were analyzed by unconditional Logistic regression models. RESULTS: The cumulative tolerance probabilities of cow's milk and egg were 42% and 31% one year after diagnosis respectively, 63% and 62% 2 years later, 77% and 80% 3 years later, and 100% after 4 years. And the severity of skin prick test reactions to cow's milk and egg was the predicting factor for persisting hypersensitivity to cow's milk and egg (OR = 2.535, 95% CI: 1.159-5.543; OR = 2.654, 95% CI: 1.302-5.410, P < 0.05). Thirteen cases presented with hypersensitivity to other foods (15.6 +/- 6.1) months after the diagnosis was confirmed, and the risk factor was the long-lasting hypersensitivity to egg (OR = 6.109, 95% CI: 1.818-20.527, P < 0.05). After 4 cases and 15 cases were diagnosed as allergic rhinitis and asthma, respectively, (16.8 +/- 8.3) months after diagnoses and the risk factors were the long-lasting hypersensitivity to egg and the respiratory symptoms (OR = 3.596, 95% CI: 1.429-9.045; OR = 4.235, 95% CI: 1.152-15.563, P < 0.05). CONCLUSION: At least 75% of children with egg or cow's milk allergy could develop tolerance to egg or cow's milk within 3 years after diagnoses; 10.9%, 12.6% and 3.4% of children with food allergy suffered from other food allergy, asthma and allergic rhinitis. Strengthening the screening and management of children at high risk for persistent food allergy will contribute to improvement of the prognoses of food allergy. PMID- 16255861 TI - [Establishing the norm of cognitive adaptive test/clinical linguistic and auditory milestone scale in the children from 4 to 36 months of age in Shanghai, China]. AB - OBJECTIVE: To meet the need for instrument assessing the cognitive abilities of infants and young children as well as discriminating between global developmental delay and particular deficits in either language or problem-solving skills, we intended to introduce Cognitive Adaptive Test/Clinical Linguistic and Auditory Milestone Scale (CAT/CLAMS) into China. METHODS: CAT/CLAMS were administered to 1604 normative children aged 4-36 months (in 16 age groups, about 100 children per age group) in Shanghai during the period from December 2003 to June 2004. In the meantime, Gesell Developmental Diagnosis was applied for 100 of these children, respectively aged 4, 6, 12, 18 and 30 months (20 children per age group). Interclass correlation coefficients (ICC) were adopted to analyze data in terms of inter-rater reliability and re-test reliability of the scales of CAT/CLAMS. Cronbach alpha coefficients were calculated to assess the inter consistency of the scales. Pearson correlation coefficients(r) were adopted to analyze the concurrent validity of the scales. The normative percentile graphs of CAT/CLAMS in the children from 4 to 36 Months of age in Shanghai, China were adopted. RESULTS: Administrations of the CAT/CLAMS for each subject usually took 10-20 minutes. Individual scores (CLAMS, CAT, and CAT/CLAMS) increased with ages (Pearson correlation coefficients were 0.96, 0.98 and 0.98, respectively, P < 0.01 for all). ICCs (intraclass correlation coefficient) in terms of individual scores for the inter-rater reliability test and the re-test reliability test were respectively > or = 0.96 (P < 0.01) and > or = 0.95 (P < 0.01), all the Cronbach alpha coefficients were > or = 0.98; in 100 children of the 5 age groups, there was significantly positive correlation between CAT/CLAMS and Gesell Developmental Diagnosis in terms of language skill DQ and adaptive skill DQ, and Full Scale DQ (r = 0.517, 0.703, 0.613, respectively, P < 0.01 for all). Moreover, this significant positive correlation was observed in each of the 5 age groups (r = 0.455-0.827, P < 0.05). CONCLUSION: CAT/CLAMS is suitable for discriminating between global developmental delay and particular deficits in either language or problem-solving skills. It is a quick, reliable, and valid instrument, with refined and quantified results. It is a good tool for developmental surveillance and screening of infants and young children. PMID- 16255862 TI - [Experimental study on the effects of nitric oxide inhalation on formation of lung nitrotyrosine and apoptosis of alveolar cells]. PMID- 16255863 TI - [Changes of calpain in renal tubular epithelial cells during kidney ischemia/reperfusion injury of neonatal rats]. PMID- 16255864 TI - [Clinical characteristics of Mallory-Weiss syndrome in children]. PMID- 16255865 TI - [Both social and economic benefits should be considered simultaneously in prevention and treatment of hereditary metabolic diseases]. PMID- 16255866 TI - [Feasibility and economic issues should be considered for setting up of research projects for screening in population]. PMID- 16255867 TI - [A brief description of 2004 national conference of the American Academy of Pediatrics]. PMID- 16255868 TI - [Neonatal hepatic focal nodular hypertrophy in a case]. PMID- 16255869 TI - [A case of chronic diarrhea and hypoproteinemia associated with Cryptosporidium parvum and Cyclospora cayetanenis]. PMID- 16255870 TI - [A child with progressive pigmented purpuric dermatosis]. PMID- 16255871 TI - [Application of hematopoietic stem cell transplantation in non-hematologic diseases]. PMID- 16255872 TI - [Critical timing for standardization of internal medicine training]. PMID- 16255873 TI - [Supplement but not replacement]. PMID- 16255874 TI - [Detection of Aspergillus infection in blood of malignant hematopoietic tumor patients by two-step polymerase chain reaction]. AB - OBJECTIVE: To investigate the sensitivity and specificity of two-step PCR in detection of Aspergillus species in blood from malignant hematopoietic tumor patients. METHODS: Forty-one blood samples from high-risk patients were detected with two-step PCR. The clinical applicability was assessed with analyzing computed tomography, Aspergillus culture, neutropenia and outcome after anti fungal therapy. RESULTS: Specific band of PCR was obtained from Aspergillus strains and products of PCR had a high homogeneous in sequence with Aspergillus species. Sixteen of 41 patients were PCR positive and controls were all PCR negative. Among PCR positive patients, 4 patients had positive cultures. The lowest WBC count was (0.30 +/- 0.14) x 10(9)/L and (0.50 +/- 0.26) x 10(9)/L respectively and duration with WBC count less than 1.0 x 10(9)/L was (19.00 +/- 8.31) days and (12.69 +/- 6.95) days respectively in the two groups. After antifungal therapy, one of the patients with PCR positive died of chemotherapy related early death. CONCLUSIONS: The two-step PCR assay allows for highly sensitive and specific detection of Aspergillus pathogens in vitro and in vivo. The detection with PCR has some value in the early diagnosis and helps us to make decision for antifungal therapy. PMID- 16255875 TI - [A clinical pharmacokinetic study of multi-dose oral tetra-arsenic tetra-sulfide combination therapy in acute promyelocytic leukemia]. AB - OBJECTIVE: To study the pharmacokinetics of multi-dose oral tetra-arsenic tetra sulfide (As(4)S(4)) in acute promyelocytic leukemia (APL) patients and its major side effects. METHODS: Clinical pharmacokinetic study of As(4)S(4) was carried out in 7 patients who had never used arsenic before. The total concentration of arsenic in blood, urine and hair was determined with hydride generation-atomic absorption spectrometry. RESULTS: These patients were administrated oral As(4)S(4) complex capsule 20 mg/kg three times a day for 14 days. From day 10 and to day 14, the blood arsenic concentration and urinary arsenic excretion reached a steady state. The average minimal concentration (Cmin) and maximal concentration (Cmax) of blood arsenic was (53.3 +/- 9.0) microg/L and (70.7 +/- 10.8) microg/L. t 1/2 was prolonged to (70.7 +/- 31.7) hour, and area under curve (AUC) was (448.9 +/- 71.8) microg. h(-1).L(-1). Median time to peak concentration (Tmax) was 1 (range 0.5 - 8) hour. During As(4)S(4) therapy, 24-hour arsenic content in urine was (6170.8 +/- 3141.8) microg/L and it accounted for about (0.152 +/- 0.082)% of the total daily dosage. It decreased steadily over time after drug withdrawal, arsenic accumulated in hair and the concentration could be ten-fold higher than that before treatment. CONCLUSION: Multi-dose oral As(4)S(4) is safe and has been relatively well tolerated in APL patients in spite of the tendency of its retention in some tissues after long time administration. PMID- 16255877 TI - [A clinical trial of anti-CD4 CD8 monoclonal antibodies combined with cyclosporine A in treatment of severe aplastic anemia]. AB - OBJECTIVE: To explore the safety and efficacy of a therapeutic regimen for treating severe aplastic anemia (SAA) in its early stage. METHODS: Two groups of SAA patients were enrolled in the present study. One was a treatment group including 21 patients being treated with anti-CD(4), CD(8) monoclonal antibodies as well as cyclosporine A. Another was a control group including 20 patients being treated with anti-lymphocyte globulin and cyclosporine A. RESULTS: The response rates in the two groups were more or less some, being 76.2% for the treatment group and 65.0% for the control group (P > 0.05), but the blood routine examination results showed quicker recovery in the treatment group than in the control group (P < 0.05) and the incidences of side effects related to therapy such as fever were less in the treatment group than in the control group (P < 0.01). CONCLUSIONS: The combination of anti-CD(4), CD(8) monoclonal antibodies and cyclosporine A is safe and efficient in treating SAA. PMID- 16255878 TI - [The effect of early invasive strategy on early and late outcomes in high-risk non-ST-segment elevation acute coronary syndromes]. AB - OBJECTIVE: To investigate the effect of early invasive strategy on early and late outcomes in high-risk patients with non-ST-segment elevation acute coronary syndromes (ACS). METHODS: Five hundred and forty-five patients of ACS without ST segment elevation were randomly assigned to an early conservative strategy (284 cases) or early invasive strategy group (261 cases), who were enrolled consecutively from Oct., 2001 to Oct., 2003. The combined cardiovascular events (a combination of cardiac death, nonfatal myocardial infarction, nonfatal heart failure and hospital readmission due to recurrent ischemic angina) within 30 days and 6 months were analyzed and the effects of early invasive strategy on early and late outcomes in high-risk patients with increased TnI or hs-CRP levels were evaluated. RESULTS: As compared with early conservative strategy, early invasive strategy lowered the rate of hospital readmission due to recurrent ischemic angina of 30 d and the combined cardiovascular events of 30 d and 6 months (all P < 0.05). Subgroup analysis indicated early invasive strategy could significantly decrease the incidences of the combined cardiovascular events of 30 d and 6 months and the hard end point events of 6 months in patients with increased TnI or hs-CRP levels (all P < 0.01), but no such changes could be seen in patients with normal TnI or hs-CRP levels, as compared with early conservative strategy. CONCLUSIONS: Early invasive strategy decreases significantly cardiovascular events and improves the early and late outcomes in high-risk patients with increased TnI or hs-CRP levels. PMID- 16255879 TI - [The diagnostic value of serum carcinoma markers, fecal K-ras and p53 gene mutation in pancreatic cancers]. AB - OBJECTIVE: To evaluate the diagnostic value of serum carcinoma markers CA19-9, CA242, CA50 and carcinoembryonic antigens (CEA), fecal K-ras and p53 gene mutation in pancreatic cancer. METHODS: We collected 136 new cases of pancreatic cancer and 240 patients with benign digestive diseases including 49 patients with benign pancreatic diseases diagnosed in Peking Union Medical College Hospital, Chinese Academy of Medical Sciences Tumor Hospital and Shenyang PLA General Hospital from February 2002 to March 2004. Blood samples were collected and serum carcinoma markers CA19-9, CA242, CA50 and CEA were measured. Fecal K-ras and p53 gene mutation were also measured. We decided the optimal cut-off points with receiver operating characteristic curves and calculated the areas under the curve (AUC). RESULTS: The AUC of serum CA19-9 and CA242 were 0.855 +/- 0.031 (95% CI 0.794 - 0.916) and 0.859 +/- 0.031 (95% CI 0.799 - 0.920) respectively. The optimal cut-off point for serum CA19-9 was 68 U/ml, with the sensitivity of 84.4% (98/116) and the specificity of 84.3% (145/172) for the diagnosis of pancreatic cancer. The optimal cut-off point for serum CA242 was 25 U/ml, with the sensitivity of 88.4% (84/95) and the specificity of 79.1% (144/182). The sensitivity of fecal K-ras mutation was 77.8%, and the specificity was 82.2%. The sensitivity and specificity of fecal p53 gene mutation were 27.8% and 95.2% respectively. The diagnostic scale of pancreatic cancer was calculated by four variables: serum CA19-9, CA242, fecal K-ras and p53, of which each variable deserved one point if measured positive. The optimal cut-off point for the scale was 2, and the AUC of the diagnostic scale was 0.946 +/- 0.017 (95% CI 0.912 - 0.980). CONCLUSIONS: Serum CA19-9 and CA242 are valuable diagnostic tools for pancreatic cancer. The diagnostic value will be further improved when they are combined with the measurement of fecal K-ras and p53 gene mutation. PMID- 16255881 TI - [The diagnostic value of telomerase activity in differentiating benign and malignant ascitic fluid]. AB - OBJECTIVE: To investigate the changes of telomerase activity in benign and malignant ascites fluid. METHODS: The technique of telomerase TRAP-PCR-ELISA was employed to detect telomerase activity in ascites cells from 60 patients with benign or malignant ascites fluid, the cytological diagnosis and total tumor marks (carcinoembryonic antigen, et al.) were compared. RESULTS: Telomerase activity in malignant ascites fluid was significantly higher than that in benign group. Positive rate of telomerase activity detected in malignant ascites (90%) was remarkably higher than those in benign group (10%); Compared with the cytological diagnosis and total tumor marks (carcinoembryonic antigen, et al.), telomere ferment test had higher sensitivity (90%), specificity (100%) and stability. CONCLUSION: Telomerase activity may be an useful and sensitive mark in differential diagnosis of benign and malignant ascites fluid. PMID- 16255882 TI - [The role of mitochondrial damages in Helicobacter pylori-induced apoptosis of gastric cancer cells]. AB - OBJECTIVE: To explain the role of mitochondrial pathway in the apoptosis of SGC 7901 cell line induced by concentrated Helicobacter pylori culture supernatant (CHCS). METHODS: Cytochrome oxidase (COX) I expression was detected by Western blotting. Cell apoptosis and mitochondrial membrane potential were measured by flow cytometry. RESULTS: CHCS could induce the apoptosis of SGC-7901 in a dose- and time-dependent manner. Apoptotic rates gradually enhanced followed by the concentrations increasing. The mitochondrial membrane potential (MMP) began to descend after treating CHCS for 4 h, and MMP descended most distinctly in 8 h. It descended the lowest point in 12 h, and it had no special changes in 24 h. The expression of COX I was notably lower than that of control group after CHCS treating (632.8 +/- 40.6 vs 895.1 +/- 44.2, P < 0.05). CONCLUSION: Mitochondrial pathway may play an important role in the apoptosis of SGC-7901 cells induced by CHCS. PMID- 16255883 TI - [Clinical analysis of 25 patients with bilateral pheochromocytomas]. AB - OBJECTIVE: Bilateral pheochromocytomas are rare diseases. The purpose of this retrospective study was to elucidate the clinical characteristics of patients with bilateral pheochromocytomas. METHODS: We analyze the clinical data of 25 patients with bilateral pheochromocytomas who were treated at Peking Union Medical College Hospital between 1952 and 2004. RESULTS: The average age at diagnosis was (32 +/- 14) years. 19 cases (76%) were familiar type, and among the 19 cases, 13 cases were multiple endocrine neoplasia (MEN), 5 cases von Hippel Lindau (vHL) disease and 1 case of isolated familial pheochromocytomas. In the 25 patients, bilateral pheochromocytomas were discovered at the same time in 88%, and multiple tumors existed in at least one side of the adrenal gland in 56%. 50% of cases recurred after resection of pheochromocytomas. CONCLUSIONS: Hereditary syndromes should be screened when pheochromocytoma is bilateral, and the patients' family members also should be screened for hereditary syndromes. During operation for bilateral adrenal pheochromocytoma, multiple tumors in one side should be considered. Long-term follow-up is necessary because recurrence may develop many years after operation. PMID- 16255885 TI - [The relation between fasting plasma glucose concentrations and insulin resistance]. AB - OBJECTIVE: To study whether insulin resistance exists in the Chinese individuals with impaired glucose regulation (IGR) defined by the modified cut-point value (5.6 mmol/L) in 2003 version of ADA criteria for impaired fasting glucose (IFG). METHODS: We selected 9 persons [aged (44 +/- 5) years, body mass index (BMI) (24.0 +/- 0.8) kg/m(2)] with normal glucose regulation [NGR, fasting plasma glucose (FPG) < 5.6 mmol/L, plasma glucose-2h (PG2h) < 7.8 mmol/L], 9 [aged (49 +/- 3) years, BMI (29.0 +/- 0.8) kg/m(2)] with isolated IFG defined by the new criteria (n-i-IFG, FPG < 5.6 mmol/L, PG2h < 7.8 mmol/L), 20 with combined impaired glucose tolerance (c-IGT) including 10 [aged (45 +/- 4) years, BMI (26.0 +/- 1.3) kg/m(2)] defined by the new criteria (n-c-IGT, FPG 5.6 - 6.0 mmol/L, PG2 h 7.8 - 11.0 mmol/L) and 10 [aged (49 +/- 3) years, BMI (27.0 +/- 1.1) kg/m(2)] defined by the old criteria (o-c-IGT, FPG 6.1 - 6.9 mmol/L, PG2h 7.8 - 11.0 mmol/L), 10 [aged (43 +/- 5) years, BMI (29.0 +/- 2.8) kg/m(2)] with type 2 diabetes mellitus (DM). Insulin resistance of the subjects was measured by using hyperinsulin-euglycemic clamp in which the glucose infusion rate (GIR) was the major index. Beta-cell function was assessed by intravenous glucose tolerance test. The DM patients only underwent the hyperinsulin-euglycemic clamp. RESULTS: (1) The GIR of n-i-IFG group and n-c-IGT group was significantly lower than that of the NGR group [(7.2 +/- 0.8), (7.0 +/- 1.5) vs (10.3 +/- 0.9) mg.kg(-1).min( 1), P < 0.05]. The GIR of o-c-IGT group was the least [(4.8 +/- 0.4) mg.kg( 1).min(-1)], which was similar to that of the DM group [(5.6 +/- 1.0) mg.kg( 1).min(-1)], the GIR of the two groups was significantly reduced compared with the NGR goup [(4.8 +/- 0.4), (5.6 +/- 1.0) vs (10.3 +/- 0.9) mg.kg(-1).min(-1), P < 0.01]. (2) As compared with the NGR group, the fasting insulin level was increased in all the IGR groups and was decreased in the DM group. (3) The first phase of insulin secretion were similar in NGR and n-i-IFG group [(37.0 +/- 8.6) vs (31.7 +/- 9.2) mU/L, P > 0.05], and the n-i-IFG group had greater amount than the n-c-IGT group [(31.7 +/- 9.2) vs (25.9 +/- 9.8) mU/L, P > 0.05]. But it was the least in the o-c-IGT group that significantly less than the NGR group [(17.2 +/- 4.8) vs (37.0 +/- 8.6) mU/L, P < 0.05]. As compared with the NGR group, the second phase of insulin secretion was slightly increased in the n-i-IFG group [(16.6 +/- 2.9) vs (21.9 +/- 3.6) mU/L, P > 0.05], and was even greater in the n c-IGT group [(16.6 +/- 2.9) vs (38.7 +/- 13.7) mU/L, P < 0.05]. But it was reduced in the o-c-IGT group [(20.7 +/- 2.9) mU/L]. CONCLUSIONS: (1) The IGR subjects defined by the new cut-point value already has insulin resistance. (2) Deficit in insulin secretion is worsening as the IGR deteriorates. PMID- 16255886 TI - [Comparison of oxygen therapy with nasal continuous positive airway pressure on Cheyne-Stokes respiration in patients with chronic congestive heart failure]. AB - OBJECTIVE: To compare the acute effects of oxygen therapy and nasal continuous positive airway pressure (nCPAP) therapy on Cheyne-Stokes respiration (CSR) in patients with stable chronic congestive heart failure (CHF). METHODS: Prior to the study, all patients had an echocardiogram performed to measure the left ventricular ejection fraction (LVEF). In addition, all patients had an initial sleep study to identify the presence of CSR. Those patients identified as having CSR were randomized to a night on 2 L/min oxygen therapy, a night on 4 L/min oxygen therapy (by nasal cannula) and another night on nCPAP therapy [mean pressure (9.1 +/- 1.1) cm H2O]. RESULTS: Twenty-six patients stable CHF, with a mean age of 64.1 +/- 6.8, and a mean LVEF of (27.1 +/- 5.8)%, were studied, of whom 14 (53.8%) had CSR during their initial sleep study. The 14 patients had an average apnea-hypopnea index (AHI) of 34.9 +/- 8.2 events per hour, an average apnea-hypopnea length of (20.6 +/- 3.2) s, mean cycle length (74.8 +/- 21.3) s, circulation time (25.6 +/- 4.4) s, the lowest oxygen saturation during the night (76.2 +/- 4.7)%, the periods of time with a oxygen saturation of < 90% of total sleep time (20.9 +/- 8.6)%. When compared with baseline measurements, both oxygen therapy (2 L/min or 4 L/min) and nCPAP therapy significantly decreased the AHI, with 4 L/min oxygen therapy and nCPAP therapy producing the better results, with no significant difference between these two therapies. All three forms of treatment significantly increased the lowest oxygen saturation during the night to a similar extent. The mean percent time the oxygen saturation was < 90% also improved with all interventions, with 4 L/min O(2) producing the best results. In addition, 2 L/min or 4 L/min oxygen therapy and nCPAP produced similar improvements in total sleep time and sleep efficiency. When compared with baseline measurements, the apnea-hypopnea length, cycle length, and circulation time did not significantly change with either oxygen therapy or nasal CPAP therapy. CONCLUSION: CSR occurs frequently in patients with stable CHF. Both higher concentration oxygen and nCPAP can be used as therapeutic strategies in CHF patients with CSR. PMID- 16255888 TI - [Expression and role of integrin-linking kinase in renal tubulointerstitial lesion of aging rats]. AB - OBJECTIVE: To explore the expression and role of integrin-linking kinase (ILK) in aging rats subjected to unilateral ureteral obstruction (UUO). METHODS: The UUO model and sham-operated rats (SHAM) were established with 3-month-old rats and 26 month-old rats and the rats were sacrificed before UUO and after UUO at 3, 7 and 14 days. Immunofluorescent staining, Western blot, RT-PCR, etc were applied to detect the expressions of ILK and fibronectin (FN) in the renal of UUO rat at each time point. RESULTS: With the time of UUO, the relative interstitial areas and tubulointerstitial fibrotic areas in 26-month-old rats were significantly higher than those in 3-month-old ones at each time point (P < 0.01) and the mRNA and protein levels of ILK and FN are increasing with the time of UUO in both groups (P < 0.01). ILK mRNA semi-quantitative values of 26-month and 3-month rats were 0.98 +/- 0.06 vs 0.72 +/- 0.06, 3 days after UUO (P < 0.01), 1.49 +/- 0.05 vs 1.03 +/- 0.04, 14 days after UUO (P < 0.01), respectively. ILK proteinsemi quantitative values of 26-month and 3-month rats were 0.57 +/- 0.04 vs 0.52 +/- 0.03, 3 days after UUO (P < 0.01), 0.76 +/- 0.04 vs 0.63 +/- 0.03, 14 days after UUO (P < 0.01), respectively. The expressions of ILK mRNA and protein level were positively correlated with the relatively renal interstitial areas (r = 0.71, P < 0.05 and r = 0.80, P < 0.05, respectively). And the whole levels of ILK and FN in 26-month-old rats are higher than those of 3-month-old rats. CONCLUSION: With aging, the expressions of ILK and FN increase progressively in UUO rat, and the over-expression of ILK probably promotes renal tubulointerstitial fibrosis and aging. PMID- 16255908 TI - [Clinical significance of dual-axis diagnostic criteria and the necessity of treatment for temporomandibular disorders]. PMID- 16255889 TI - [Renoprotective effect of iptakalim hydrochloride in hypertension]. AB - OBJECTIVE: To investigate the experimental therapeutic effects of iptakalim hydrochloride (Ipt) on renoprotection in spontaneously hypertensive rats. METHODS: 30 SHR were treated ig with Ipt 1, 3, 9 mg.kg(-1).d(-1), benazepril 3 mg.kg(-1).d(-1) once a day for 12 weeks. Age-matched WKY rats were used as normal control. The blood pressure, heart rates, proteinuria were assessed, and renal tissues were examined by light microscopy. The levels of blood and renal tissue ET-1 and TGF-beta1 were detected respectively by radioimmunoanalysis and enzyme linked immune absorption assay (ELISA). RESULTS: During 12 weeks experimental period, the systolic blood pressure (SBP) and heart rates (HR) of the untreated SHR were increased progressively. Ipt (3, 9 mg.kg(-1).d(-1)) could decrease effectively and inhibit the increasing tendency of HR. In addition, Ipt (1, 3, 9 mg.kg(-1).d(-1)) reduced urinary proteinuria, alleviated obviously the small vascular remodeling of renal and decreased the levels blood and renal ET-1 and TGF-beta1. Ipt (3, 9 mg.kg(-1).d(-1)) alleviated obviously the small vascular remodeling of renal compared with Ipt (1 mg.kg(-1).d(-1)). CONCLUSIONS: Ipt (1, 3, 9 mg.kg(-1).d(-1)) decreased SBP and protected the kidney of SHR. The renoprotection of Ipt may be involved in inhibiting of blood and renal tissue ET 1 and TGF-beta1. PMID- 16255909 TI - [Neuronal activities related to right-sided mastication detected with functional magnetic resonance imaging]. AB - OBJECTIVE: To observe neuronal activities related to right-sided mastication in cortex and to explore the effect of mastication on brain function. METHODS: Neuronal activities related to right-sided mastication were detected with functional magnetic resonance imaging in 10 healthy volunteers. Image data was analyzed by SPM99 software with the use of t test. RESULTS: Localization of activation in brain was different among subjects. Brain activation during mastication included left basal ganglion, Broca's area, angular gyrus and right supramarginal gyrus. CONCLUSIONS: The functional areas related to right-sided mastication and speech were closely connected, which indicated close relationship between mastication and speech. The central control of mastication might share part of common neural mechanism with that of speech. The roles of activated brain regions in mastication still remained to be elucidated. PMID- 16255910 TI - [Psychological status in patients with temporomandibular disorders]. AB - OBJECTIVE: To evaluate levels of psychosocial incapacity in patients with temporomandibular disorders (TMD) by using the Symptom Checklist 90 (SCL-90). METHODS: Three hundred and thirty-eight patients who came to the Center for TMD and Orofacial Pain of Peking University School of Stomatology, from March to September of 2004 were included in the present study. All of them were evaluated by SCL-90. The one-sample t-test and multivariate analysis of variance were performed to find the psychological difference between the normal and TMD groups. RESULTS: (1) There was a statistically significant difference (P < 0.05) in the scores of Symptom Checklist 90 between healthy controls and TMD group, and TMD patients got higher scores on somatization, obsessive compulsive, anxiety, hostility, phobic anxiety, psychoticism. (2) Twenty-three percent of 338 TMD patients had some psychological symptoms. (3) All patients were evaluated by analysis of reliability, and the reliability coefficients was 0.958. CONCLUSIONS: The psychological problems of TMD patients should be emphasized and SCL-90 instrument was also suitable to screen the psychological symptoms of TMD patients in China. PMID- 16255911 TI - [Identification and characterization of synovial mesenchymal stem cells in temporomandibular joint]. AB - OBJECTIVE: To identify and characterize mesenchymal stem cells from synovial membrane of temporomandibular joint in vitro. METHODS: Synovial mesenchymal stem cells (SMSCs) were obtained by limited dilution method and expanded in 25 ml flasks. Methyl thiazolyl tetrazolium (MTT) method was used to determine the cell growth cycles. The expressions of vimentin and keratin were respectively detected with immunocytochemistry, while the expressions of CD8, CD34, CD44, vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) were determined by flow cytometry. RESULTS: Pure mesenchymal stem cells were of spindle shape and uniform in size, which were intensively positive in vimentin, but negative in keratin. The expression of CD44, VCAM-1 and ICAM-1 were also verified by flow cytometry. CONCLUSIONS: Mesenchymal stem cells could be purified from adult synovial membrane of temporomandibular joint. PMID- 16255912 TI - [Conservative-surgical-conservative cycle of the treatment for temporomandibular disorders]. PMID- 16255913 TI - [Summary of the Fifth National Conference on Temporomandibular Disorders and Occlusion]. PMID- 16255915 TI - [Management of blood vessels in the vascularized autogenous submandibular gland transfer for severe keratoconjunctivitis sicca]. AB - OBJECTIVE: To summarize the management of blood vessels in the transfer of vascularized autogenous submandibular gland. METHODS: Sixty-eight cases of vascularized transfer of autogenous submandibular gland for severe keratoconjunctivitis sicca performed from August 1999 to April 2004 were reviewed. Data collected for each patient included general items, donor and recipient vessels, vessel crisis, salvage of the vessel thrombosis, and survival of transferred glands. RESULTS: Among the 68 cases, all the donor and recipient arteries were facial artery and superficial temporal artery respectively. For donor veins, 55 were facial veins, 1 was vein near the duct, and 12 were venae comitantes of facial artery. For recipient veins, 62 were superficial temporary veins and five were veins in the upper neck, one was deep temporary vein. Vessel thromboses occurred in 10 cases, 9 were venous and 1 were arterial. The salvage rate was 10.0%, with the overall survival rate of 86.8% of transferred gland. CONCLUSIONS: Variation in venous outflow is common in microvascular transfer of submandibular gland, and anastomosis between venae comitante of facial artery and superficial temporal vein is most dependable. Vessel anastomosis under microscope may reduce the vessel thrombosis rate after operation. PMID- 16255916 TI - [Influencing factors on bone resorption after alveolar bone grafting]. AB - OBJECTIVE: To retrospectively study alveolar autogenous bone grafting (AABG), the success rate and the factors influencing bone resorption after the surgery for the repair of alveolar cleft. METHODS: A retrospective analysis was performed in a group of 108 alveolar cleft patients receiving alveolar bone grafts. The method of regression was used for analysis. RESULTS: The overall survival rate of AABG was 88%, clinical success rate was 60%. The major factors influencing bone resorption were: infection, age, type of cleft, functional stimulation and surgeon. The factors which did not influence bone resorption were: sex, type of surgery, teeth pulling during the surgery and the width of the cleft. CONCLUSIONS: Surgery at the proper age of patients, orthodontic treatment before surgery, improvement of surgical skills, necessary functional stimulation and control of infection will improve the success rate of AABG. PMID- 16255918 TI - [Clinical and pathological analysis of oral manifestations of 40 patients with secondary syphilis]. AB - OBJECTIVE: To analyze the clinical and pathological features of 40 patients with secondary syphilis. METHODS: A total of 40 cases of secondary syphilis confirmed by serology were collected during 1994-2004 and were first diagnosed on presentation with oral lesions. RESULTS: The white patch in oral mucosa was found in 32 cases with painless or slight pain in most cases. The most common site of the lesion was the tongue. The histological examination on eight cases was initially misdiagnosed as oral candidosis or lichen planus, but confirmed as syphilis after serology revealed nonspecific inflammation with intraepithelial microabscess and dense perivascular infiltration of lymphocytes and plasma cells in connective tissue. The symptoms showed dramatic improvement in 16 cases after benzathine penicillin treatment. CONCLUSIONS: The oral manifestations of syphilis have specific clinical and pathological feature and attention should be paid to the suspicious oral lesions when patients are first presented in a dental office. PMID- 16255919 TI - [Clinicopathological study of Ewing's sarcoma/primitive neuroectodermal tumor in oral and maxillofacial region]. AB - OBJECTIVE: To study the clinicopathological features and diagnostic criteria of Ewing's sarcoma/primitive neuroectodermal tumor (Ewing's sarcoma/PNET) in oral and maxillofacial region. METHODS: There were 15 patients with Ewing's sarcoma/PNET in the last 35 years at our hospital, aged 1-49 years and mean 14.5 years. The most common manifestation was swelling of the affected region. The cases were analyzed and histological and immunohistochemical studies were also conducted to examine CD99 (12E7), Vimentin, NSE, S-100, Syn, CD45 (LCA), desmin. RESULTS: (1) The most common histological pattern of Ewing's sarcoma/PNET was a lobular arrangement of uniform, small, hyperchromatic cells in a fibrous background. Some of these tumors were rich in cytoplasmic glycogen. (2) Of the 7 cases, IHC was positive for CD99 (12E7) and Vimentin and negative for lymphoid (CD45), muscle (desmin) markers. S-100 was positive in 4 cases and NSE negative in 3, Syn positive in 1. NSE, S-100, Syn were all negative in 2 cases. CONCLUSIONS: Ewing's sarcoma/PNET is more common in teenagers and young people. Immunohistochemistry is essential to distinguish Ewing's sarcoma/PNET from other small round cell tumors. Immunohistochemistry is useful in the diagnosis. PMID- 16255921 TI - [Dynamic observation of lung metastasis model of adenoid cystic carcinoma]. AB - OBJECTIVE: To form ACC-M-GFP cells by transfecting pEGFP-1 into ACC-M cells, and to build up lung metastasis model of adenoid cystic carcinoma for dynamic observation in simulated lung environment. METHODS: pEGFP-1 was prepared and then transfected into ACC-M cell lines by using cationic lipid-based gene transfer technique. After successive selection, the ACC-M-GFP cells were collected and inoculated in BALB/C mice. The simulated lung environment was built up, and dynamic observation was performed by laser confocal microscopy within four weeks after transfection. RESULTS: The amount of fluorescent tumor cell colonies and the intensity of fluorescence gradually increased from the second week to the forth week after transfection. In the meantime, only a very small number of tumor cells had the ability to form clones. CONCLUSIONS: We successfully built up a lung metastasis model of adenoid cystic carcinoma for dynamic observation. The model is suitable for capturing and analyzing metastatic tumor cells in early stage. PMID- 16255923 TI - [p53 gene intron 7 polymorphism and its association with oral neoplasms]. AB - OBJECTIVE: To investigate the association between oral neoplasm genetic susceptibility and genetic polymorphism of p53 intron 7. METHODS: The intron 7 ApaI polymorphism of p53 was analyzed in 95 oral neoplasm patients and 105 healthy individuals by utilizing polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) genotyping assay technique, and direct sequencing was performed in 30 cases which were selected from the patients and controls by random sampling. RESULTS: In oral neoplasms cases, haplotype combinations were T G 43.2%, C-T 56.8%, and frequencies of genotype were T-G/T-G 15.8%, C-T/T-G 54.7%, C-T/C-T 29.5%, while in controls they were T-G 30.9%, C-T 69.1% and T-G/T G 10.5%, C-T/T-G 41.0%, C-T/C-T 48.5%. There was a significant difference in the allelic frequency and the genotypical distributions between the oral neoplasm patients and the controls. The individuals with the T-G allele had a slight increasing neoplasm risk than individuals with C-T allele; the OR for T-G versus C-T was 1.69 (95% CI, 1.12 - 2.51). The risk of suffering from oral neoplasms was higher in the individuals of T-G/T-G genotype and of T-G/C-T genotype than in individuals of C-T/CT genotype with odds ratio of 2.48 versus 2.20. CONCLUSIONS: There are two polymorphic points in the 7th intron of human p53 gene, which could be associated with genetic susceptibility of oral neoplasms. T-G allele may be the risk factor of oral neoplasms. PMID- 16255925 TI - [Establishment of the three-dimensional finite element model of the first permanent mandibular molar and its stress analysis]. AB - OBJECTIVE: To establish more rational three dimensional finite element model (FEM) of the first permanent mandibular molar and to study the internal stress distribution. METHODS: The FEM was applied to analyze the stress distribution of the first permanent mandibular molar under various loading conditions. RESULTS: Under all five loading conditions, the maximum von-Mises stress and compressive stress in enamel were larger than that in dentin, and the tensile stress in enamel was smaller than that in dentin. When vertical force and buccal-lingual oblique force were loaded, the stresses in the enamel and dentin were minimum; when lingual-buccal oblique force was loaded, the stresses were maximum; the stresses were mainly distributed at cervical region, furcation and apical area of the mesial-lingual root canal wall. CONCLUSIONS: The stress value changes and stress distribution in the enamel and dentin were closely related with the direction of the occlusion force. PMID- 16255924 TI - [Effects of 17 beta-estradiol on the adhesion, invasion and motility potential of salivary mucoepidermoid carcinoma Mc3 cells]. AB - OBJECTIVE: To study the effects of 17 beta-estradiol on the adhesion, invasion and motility potential of salivary mucoepidermoid carcinoma Mc3 cells. METHODS: The effects of 17 beta-estradiol on adhesion, invasion and motility potential of salivary mucoepidermoid carcinoma Mc3 cells were investigated with cell attachment assay on fibronectin (FN), wound assay, chemotaxis assay, and gelatin incorporated SDS-PAGE electrophoresis. The expression of estrogen receptor in Mc3 cells was determined by immunohistochemistry assay. RESULTS: Attachment rates of Mc3 cells treated with E2 at 10(-9), 10(-8), 10(-7), 10(-6) mol/L were 38.3%, 50.4%, 69.2% and 91.1% respectively, and the rate in control was 25.0%. When exposed to 17 beta-estradiol at 10(-9), 10(-8), 10(-7) and 10(-6) mol/L for 48 h, motility of Mc3 cells on FN increased by 16.9%, 40.9%, 36.4% and 38.8% respectively. When at 10(-6) mol/l, 17 beta-estradiol increased chemotaxis potential of Mc3 cells to FN by 60.3%. The activity of 68 000 matrix metalloproteinase (MMP-2) of Mc3 cells was enhanced at different levels by 10( 9), 10(-8), 10(-7), 10(-6) mol/L of 17 beta-estradiol, and estrogen receptor was also detected in nucleus of Mc3 cells by immunohistochemistry assay. CONCLUSIONS: 17 beta-estradiol at physiological concentration may enhance the adhesion, invasion and motility potential of salivary mucoepidermoid carcinoma Mc3 cells. PMID- 16255927 TI - [Gene expression of transcription regulator LMO 4 in tooth morphogenesis]. AB - OBJECTIVE: To investigate the expression of transcription regulator LMO4 mRNA in the developing mouse molar and compare the expression pattern of LMO4 with that of Shh signaling molecule. METHODS: Wild-type embryos used in this study (E11.5 P1.5) were generated by mating Kun-Ming mice. The expression pattern of LMO4 during organ development was carried on by whole-mount in situ hybridization. The expression patterns of LMO4 and Shh mRNA during molar development were analysed by section in situ hybridization. Immunohistochemical staining of PCNA was carried on by SP method. RESULTS: LMO4 mRNA was widespread at early embryonic stages (E11.5) with positive hybridization signal in the mandibular reason, limb bud, brain, epidermis and somites revealed by whole-mount in situ hybridization. Section in situ hybridization showed that LMO4 was expressed in the tooth bud, the two tips of the enamel organ and the cervical loop from E13.5 to E16.5. While Shh was localized in the enamel knot on E14.5. On E18.5-P1.5, LMO4 transcripts were distributed in the ameloblast and the stratum intermedium. On E13.5-E16.5, the tooth bud cells and the cervical loop cells were PCNA positive. These were the same regions that showed LMO4 mRNA expression. CONCLUSIONS: LMO4 was confined to the dental epithelium and had spatial temporal expression patterns during tooth morphogenesis. The expression patterns of LMO4 and Shh were similar. In early tooth development, LMO4 might regulate cell proliferation. In late tooth development, it might participate in the ameloblast differentiation. PMID- 16255929 TI - [A histological study of three-dimensional external zygomatic suture distraction osteogenesis]. AB - OBJECTIVE: To explore the histological change in suture of zygomatic bone for the zygomatic suture direct distraction osteogenesis. METHODS: The zygomatic bone was distracted by 3-D external distraction appliance without osteotomy. The specimens were taken 1, 3, 5 and 8 weeks after, and then examined histologically and compared with the blank contralateral side. RESULTS: There were lots of fibroblasts, osteoblasts and capillary vessels in the distracted suture tissues one week after distraction, and the fibers were observed to connect the sides of suture and arranged orderly. The surfaces of the expanded suture were irregular. Bone formation was active in the expanded side. The bone trabeculae were mature and oriented in the direction of distraction in the distracted sides at 3 weeks. A great amount of new woven bones were found in 5-week specimen. New bones were formed completely 8 weeks after the distraction. CONCLUSIONS: New bone formed rapidly in the distracted side of zygomatic bone under suture distraction osteogenesis without osteotomy. PMID- 16255930 TI - [The preliminary study of three-dimensional simulation of the craniofacial system]. AB - OBJECTIVE: To look for the way of three-dimensional simulation of the craniofacial system. METHODS: A three-dimensional laser scanner was used for gypsum models digitization and computed tomography scans was employed for skull reconstruction, then the data of teeth and temporomandibular joint were picked up and integrated. The ARCUS sigma system was used to record spatial mandibular movements. The data of both digital reconstruction and spatial movements were transferred into one coordinate system. The software for three-dimensional simulation was programmed. RESULTS: The preliminary program could be used to analyze static and dynamic occlusion and gnathic relations, to check the contact points and to show from various visual angles and slices. The occlusal plane, curves, and helical axis were initially defined and displayed. CONCLUSIONS: Using available instruments and methods, we developed the primary edition for three dimensional simulation of the craniofacial system. However, it is far from a mature system and there is still plenty of work to be done. PMID- 16255931 TI - [Tissue engineering of dentin-pulp complex-like structures by human dental mesenchymal cells]. AB - OBJECTIVE: To establish three-dimensional culture model of human dental mesenchymal cells and bioengineer in vivo with ceramic bovine bone (CBB) and Collagraft as scaffolds. METHODS: Human dental mesenchymal cells induced upon stimulation of bFGF and IGF-1 or TGF-beta(1) were implanted onto CBB and Collagraft containing the same kinds of growth factors respectively. Then cell/scaffold constructs were transplanted into nude mice to establish in vivo culture model of dental mesenchymal cells. Control groups were set up at the same time. After 4 weeks or 10 weeks, the implants were taken out for histological and immunohistochemical analysis. RESULTS: Within 10-week implant tissues, typical dentin-pulp complex-like structures were generated in scaffolds containing growth factors. Human dentin sialoprotein (DSP) was expressed in the newly formed dentin. This phenomenon wasn't observed in control groups and 4-week implants. CONCLUSIONS: Dentin-pulp complex-like structures could be bioengineered successfully with human dental mesenchymal cells and CBB or Collagrafts containing growth factors in nude mice. PMID- 16255932 TI - [Use of allogenic acellular dermal matrix combined with autologous epidermal cells for the repair of tissue defect]. AB - OBJECTIVE: To investigate a method for the repair of tissue defect. METHODS: Allogenic acellular dermal matrixes (ADM) were implanted to full-thickness skin defects made on the dorsa of rats. Two weeks later, autologous suspended epidermal cells were transplanted on to the surface of vascularized ADM. Respectively, neoepidermis was macroscopically observed 2, 3, 5 weeks after grafting, and samples were taken to make routine paraffin sections for microscopical examination, and immunohistochemical staining for type IV collagen was also performed. RESULTS: The vascularized ADM could support proliferation and differentiation of epidermal cells, and also could promote the formation of dermal-epidermal junction. Suspended epidermal cells in an artificial culture system in vivo could develop into mature epidermis. The reconstructed skin not only looked like the normal one in appearance in which hair was removed, but also revealed a better function. CONCLUSIONS: Full-thickness skin defect can be repaired by transplanting autologous epidermal cell suspension on to vascularized ADM. PMID- 16255934 TI - [Evaluation of the psychosocial status of fluorosis patients in fluorosis epidemic areas]. AB - OBJECTIVE: To explore the psychosocial effect of fluorosis on patients in fluorosis epidemic areas. METHODS: Multistage, stratified, clustered and random sampling method was used in this survey. Totally 416 inhabitants of 12, 15 and 35 44 years of age from rural fluorosis epidemic areas in Hebei province were selected. Dental fluorosis was classified by Dean's index (DI). A total of 178 patients with different types of fluorosis were involved in a psychosocial questionnaire investigation which included six five-point subscales, including attitude to teeth, subjective well-being, life satisfaction, interpersonal or social anxiety, effect of self-esteem, impact on behavior. RESULTS: Fluorosis prevalence in fluorosis epidemic areas of Xingtai in Hebei province was 71.20%, and significant difference existed among three age groups (Pearson Chi-square value = 167.51, P < 0.01). With the increase of DI score, the average value of subscale "attitude to teeth" rising, ranging from 2.58 to 3.51, and value of "subjective well-being" was also increasing, ranging from 2.35 to 2.9. Statistical difference of subscale average value was found only in these two subscales ("attitude to teeth": F = 4.787, P < 0.01; "subjective well-being": F = 2.538, P < 0.05). "Attitude to teeth" was significantly different between control group and moderate group (F = 0.52, P < 0.01) and between control group and severe group (F = 0.72, P < 0.01). While significant difference of "subjective well-being" could only be found between control group and severe group (F = 0.56, P < 0.01). According to the bivariate correlation analysis, strong correlation could be found between the degree of fluorosis and the average value of sub scales. CONCLUSIONS: The psychosocial impact of fluorosis on patients in fluorosis epidemic areas mainly appeared in two aspects, i.e. attitude to teeth and the perception of well-being, especially in severe group. PMID- 16255935 TI - [A study on the epidemiology of permanent tooth caries in juvenile group in Anhui province]. AB - OBJECTIVE: To survey the prevalence of dental caries and the needs for dental treatment in Anhui province. METHODS: The survey of prevalence of dental caries in group aged 12 and 15 years in Anhui province was conducted using the non proportional, stratified, cluster sampling method. RESULTS: DMFT in 12-year and 15-year group was 0.58 and 0.95 respectively, and permanent tooth caries prevalence rate was 30.78% and 39.78% respectively, lower than that of same age of other 11 provinces in mainland China (P < 0.01). The constituent ratio of DMFT was 90.51% (D), 2.48% (M), 7.01% (F) in 12-year group and 86.78% (D), 4.30% (M), 8.92% (F) in 15-year group. CONCLUSIONS: When we controlled dental caries in 19.98% 12-years crowd and 22.8% 15-years crowd of Anhui province, we will had controlled the occurrence of 80% caries too in group aged 12 and 15 years. PMID- 16255936 TI - [Surgical management of temporomandibular disc perforation]. PMID- 16255937 TI - [Clinical manifestation and treatment of occlusal diseases]. PMID- 16255938 TI - [Non-operative treatment for temporomandibular disorders]. PMID- 16255940 TI - [Rights maintenance and self-discipline of practicing dentist: Part V. Consequence in medical dispute]. PMID- 16255942 TI - [Treatment of periodontal disease: Part V. Periodontal medicine]. PMID- 16255943 TI - [Study of improving the quality of teeth and periodontal tissue united slides]. PMID- 16255944 TI - [Present studies of the relationship between taste and brain functions]. PMID- 16255945 TI - [The significance of smear-negative pulmonary tuberculosis in the control of tuberculosis]. PMID- 16255952 TI - [Early use of noninvasive positive pressure ventilation for patients with acute exacerbations of chronic obstructive pulmonary disease: a multicentre randomized controlled trial]. AB - OBJECTIVE: To assess the efficacy and safety of noninvasive positive pressure ventilation (NPPV) for acute exacerbations of chronic obstructive pulmonary disease (AECOPD) patients on general wards. METHODS: A prospective multicentre randomized controlled trial was conducted in 19 hospitals in China over 16 months. 342 AECOPD patients with pH > or = 7.25 and arterial partial pressure of carbon dioxide (PaCO(2)) > 45 mm Hg (1 mm Hg = 0.133 kPa) were recruited on general wards and randomly assigned to standard medical treatment (group A, n = 171) or early administration of NPPV plus standard medical treatment (group B, n = 171). RESULTS: The characteristics of the two groups on admission were similar. The number of AECOPD patients requiring intubations in group B was significantly less than that of the group A (8/171, 26/71, P = 0.002). Subgroup analysis showed the need for intubation in both the mild (pH > or = 7.35) and the severe (pH < 7.30) acidotic patients in group B were decreased (9/80, 2/71, P = 0.047 and 8/30, 3/43, P = 0.048 respectively). The mortality in hospital was reduced slightly by NPPV but with no significant difference (7/171, 12/171, P = 0.345). Respiratory rate (RR), scale for accessory muscle use and arterial pH improved rapidly at the first 2 hours only in patients of group B. After 24 hours, the differences of pH, arterial partial pressure of oxygen (PaO(2)), scale for accessory muscle use and RR in group B [7.36 +/- 0.06, (72 +/- 22) mm Hg, 2.5 +/- 0.9, (22 +/- 4)/min] were statistically significant compared with group A [7.37 +/- 0.05, (85 +/- 34) mm Hg, 2.3 +/- 1.1, (21 +/- 4)/min, P < 0.01 for all comparisons]. CONCLUSIONS: The early use of NPPV on general wards improves arterial blood gas and respiratory pattern, decreases the rate of need for intubation in AECOPD patients. NPPV is indicative for alleviating respiratory muscle fatigue and preventing respiratory failure from exacerbation. PMID- 16255953 TI - [Identification of a plasmid-mediated AmpC beta-lactamase from clinical isolates of Escherichia coli and Klebsiella pneumoniae]. AB - OBJECTIVE: To investigate the susceptibility and genotype characteristics of Escherichia coli and Klebsiella pneumoniae producing plasmid-mediated AmpC beta lactamase. METHODS: A total of 110 strains of Escherichia coli and Klebsiella pneumoniae were collected from patients hospitalized in our respiratory ward during January 2002-May 2004. The isolates harboring AmpC beta-lactamase were detected by three-dimensional test, and isoelectric focusing analysis, electroporation, PCR and DNA sequencing were performed to determine the genotypes. RESULTS: AmpC enzyme was detected in 9.30% (4/43) of the Escherichia coli isolates and 4.48% (3/67) of the Klebsiella pneumoniae isolates. The susceptibility test showed 7 isolates tested were all resistant to cefoxitin, and part of these strains were resistant to the third-generation cephalosporins, beta lactams/beta-lactamase inhibitors, aztreonam, amikacin and ciprofloxacin, but most of them were susceptible to cefepime and imipenem. Three strains of Klebsiella pneumoniae and 2 strains of Escherichia coli transferred cefoxitin resistance to recipients by electroporation. Results of PCR and DNA sequencing defined them to be DHA-1 AmpC beta-lactamase. CONCLUSION: Escherichia coli and Klebsiella pneumoniae producing plasmid-mediated AmpC beta lactamase were isolated from clinical strains in this hospital, and its gene can be transferred horizontally. PMID- 16255954 TI - [Therapeutic effect of ceftazidime in a rabbit model of peritonitis caused by Escherichia coli producing CTX-M-14 extended-spectrum beta-lactamase]. AB - OBJECTIVE: To study the therapeutic effect of ceftazidime (CAZ) in a rabbit model of peritonitis caused by Escherichia coli producing CTX-M-14 extended-spectrum beta-lactamase (ESBL). METHODS: The peritonitis model was produced by intra abdominal injection of a mixture of 3 x 10(8) CFU/ml Escherichia coli producing CTX-M-14 ESBL and 10% BaSO(4). All rabbits were randomized into four groups. Four hours after the bacteria were injected, three groups were intramuscularly administered ceftazidime (100 mg/kg, twice a day), cefotaxime (100 mg/kg, twice a day) and piperacillin/tazobactam (225 mg/kg, every 8 h) respectively. The fourth group did not receive any therapy as a control. The temperature, leucocyte counts and percentage of neutrophils were closely observed. The dying time of rabbits was recorded and anatomized immediately, and pathology of the great omentum was examined. RESULTS: The temperature of the control group became normal on the 6th day, while the temperature of the other three groups returned normal 48 h after the injection of the bacteria. Compared with the mortality of the cefotaxime treated group (50%) and the control group (52%), the mortality of the ceftazidime treated group (22%) was significantly decreased (chi(2) = 5.64, 6.13, P < 0.05). The administration of ceftazidime also inhibited the incidence of abdominal abscess in the survived rabbits (16%) in comparison with the control group (50%, chi(2) = 3.93, P < 0.05). In addition, the leucocyte counts of the ceftazidime treated group returned normal at the end of treatment, while those of the cefotaxime group and the control group were still high. The above measurements showed no difference between the ceftazidime-treated group and the piperacillin/tazobactam-treated group (P > 0.05). CONCLUSION: Ceftazidime showed good therapeutic effect in peritonitis caused by Escherichia coli producing CTX-M 14 ESBL. PMID- 16255955 TI - [Endogenous hydrogen sulfide in patients with chronic obstructive pulmonary disease]. AB - OBJECTIVE: To investigate the role of endogenous hydrogen sulfide (H(2)S) in patients with chronic obstructive pulmonary disease (COPD). METHODS: Levels of serum H(2)S and nitric oxide (NO), lung function and cell differential count in induced sputum were studied in 27 patients with acute exacerbation of COPD (AECOPD), 37 patients with stable COPD and 13 health subjects. Echo-Doppler assessment and arterial blood gas were measured in patients with AECOPD. RESULTS: (1) The serum H(2)S level was significantly higher in patients with stable COPD [(50.8 +/- 2.5) micromol/L] as compared to those in the controls [(39.8 +/- 1.6) micromol/L] and in patients with AECOPD [(33.5 +/- 2.2) micromol/L, P < 0.01]. (2) The level of serum H(2)S was significantly lower in smokers with AECOPD [(28.1 +/- 1.3) micromol/L] as compared to nonsmokers with AECOPD [(39.4 +/- 3.9) micromol/L, P < 0.05] and healthy nonsmokers [(39.8 +/- 1.6) micromol/L, P < 0.01]. (3) There was significant difference in the serum H(2)S level among stable COPD patients with different severity of airway obstruction (P < 0.05); being lower in patients with stage III [(45.1 +/- 4.1) micromol/L] as compared to stage I obstruction [(70.2 +/- 6.2) micromol/L, P < 0.05]. (4) AECOPD with pulmonary hypertension pulmonary artery systolic pressure (PASP) > or = 35 mm Hg (1 mm Hg = 0.133 kPa) showed a lower serum H(2)S level [(26.3 +/- 2.2), (36.2 +/- 2.5) micromol/L, P < 0.05] than that with a normal resting PASP. (5) H(2)S in serum was positively correlated with NO levels (r = 0.278, P = 0.029), FEV(1)% predicted values (r = 0.533, P = 0.000), percentage of sputum lymphocytes (r = 0.286, P = 0.028) and macrophages (r = 0.334, P = 0.01); and negatively correlated with PASP (r = -0.561, P = 0.011) and the percentage of sputum neutrophils (r = -0.422, P = 0.001) in patients with COPD. CONCLUSION: Endogenous H(2)S may be involved in the pathogenesis of airway obstruction in COPD and may be a noninvasive marker of disease activity and severity. PMID- 16255956 TI - [Effects of airway epithelium injury on the transdifferentiation of sub epithelial fibroblasts and its role in the development of airway hyperresponsiveness in asthma]. AB - OBJECTIVE: To address the possible role of injured airway epithelium in initiating transdifferentiation of sub-epithelial fibroblasts into myofibroblasts and accelerating cell proliferation in sub-epithelial fibroblasts, which may be involved in airway hyperresponsiveness in asthma. METHODS: Human primary cultured sub-epithelial fibroblasts were co-cultured with human bronchial epithelial cells (16HBE) which were treated with lipopolysaccharide (LPS) plus mechanical scratch prior to co-culture. The procedure was also performed in the presence or absence of endothelin (ET) receptor A inhibitor (BQ123), transforming growth factor beta(1) (TGF-beta(1)) neutralized antibody, respectively or simultaneously, followed by immunostaining, Western blotting and bromodeoxyuridine (BrdU) incorporation respectively to detect alpha-SMA expression and cell proliferation in the co-cultured sub-epithelial fibroblasts. Using the inhibitors specific for mitogen-activated protein kinases (MAPKs) pathways, the role of MAPKs pathways in activating the expression of alpha-SMA was evaluated. In addition, the interaction between matrix metalloproteinases (MMPs) and ET-1 was investigated by cell transfection with anti-ET-1 converting enzyme (anti-ECE) mRNA expression plasmid followed by gelatin zymography analysis. RESULTS: 16HBE treated with LPS plus mechanical injury induced alpha-SMA expression in sub-epithelial fibroblasts and accelerated BrdU incorporation in the cells. BQ123, TGF-beta(1) neutralized antibody, specific inhibitors for p38 MAPK and extracellular signal-regulated kinase 1/2 (ERK1/2) were able to block the induction respectively to a certain extent. Phosphorylated p38 MAPK and ERK1/2 were detected in the sub-epithelial fibroblasts 10 min after being co-cultured with injured 16HBE. Compared to normal control (16HBE transfected with pEGFPN(2)) or those cells transfected with anti ECE mRNA expression plasmids, ET-1 released from the 16HBE cells transfected with pEGFPN(2) into supernatants were increased significantly after the treatment described as above: 16HBE pre-transfected with pEGFP-N(2) expression plasmid before being treated with mechanical scrape plus LPS stimulation: (15.00 +/- 0.86) pg/ml; 16HBE pre-transfected with anti-ECE expression plasmid before being treated with mechanical scrape plus LPS stimulation: (7.57 +/- 0.94) pg/ml (all P < 0.01). At the same time, the activities of MMP-2 and MMP-9 were enhanced. CONCLUSIONS: Injured airway epithelial cells induced the transdifferentiation of sub-epithelial fibroblasts into myofibroblasts, which may be mediated by ET-1 and TGF-beta(1) through MARKs pathways such as p38 MAPK and ERK 1/2. PMID- 16255957 TI - [The value of detection of gene rearrangement of immunoglobulin heavy chain and immunohistochemistry in pulmonary mucosa-associated lymphoid tissue type lymphoma]. AB - OBJECTIVE: To investigate the features of gene rearrangement of immunoglobulin heavy chain (IgH) and immunophenotypes of pulmonary mucosa-associated lymphoid tissue type lymphoma (MALTLoma). METHODS: The clinical and pathological data of 12 cases with pulmonary MALTLoma and follow-up information were retrospectively reviewed, and the paraffin-embedded samples were examined with immunohistochemistry staining (12 cases) and semi-nested polymerase chain reaction (PCR) for IgH and T-cell receptor gamma (TCRgamma) gene rearrangement (7 cases). RESULTS: The patients included 9 cases confirmed by open or video assisted thoracoscopic lung biopsy and 3 cases by needle lung biopsy. Histopathologically, the tumors were composed of a spectrum of cell types that included mainly centrocyte-like cells and small lymphocytes. Lymphoepithelial lesions were identified in 12 cases, reactive colliculus lymphaticus in 11 cases, follicular colonization in 10 cases, vascular infiltration in 9 cases, and pleura involvement in 4 cases. All cases showed immunoreactivity for B-cell correlative markers. Positivity for FR2 and FR3A primers in MALTLoma were found in 6 case and 5 case respectively. The detection of TCRgamma1 and TCRgamma2 was negative in 7 cases. The combined positive rate was 100%. Chemotherapy alone was administered in 3 patients, surgery alone was performed in 8 patients, and chemotherapy after operation was carried out in 6 patients. Follow-up data were available in 11 patients. Eight of them were alive and stable, one experienced relapse and two died of the disease within 11 years and 12 years after diagnosis respectively. CONCLUSIONS: Most of the cases of pulmonary MALTLoma can be diagnosed with morphology and immunohistochemistry staining if the lesions are typical. PCR detection of IgH gene rearrangement would be helpful in differential diagnosis from benign lymphoplasia of the lung. PMID- 16255958 TI - [The effect of STAT1 antisense oligonucleotides on lung fibroblast proliferation and hydroxyproline secretion]. AB - OBJECTIVE: To investigate the effect of signal transducer and activator of transcription 1 (STAT1) antisense oligonucleotides on lung fibroblast proliferation and hydroxyproline secretion. METHODS: Ten adult female Wistar rats were randomly divided into two groups: one group was intratracheally instilled with bleomycin (BLM), while another group with 0.9% NaCl solution (NS). After 7 days, the rats were killed by right ventricle of heart exsanguinations under ketamine anaesthesia, and bronchoalveolar lavage (BAL) was performed to obtain alveolar macrophage (AM). AMs from the BLM group were divided into four groups, treated with STAT1 antisense oligonucleotides, STAT1 sense oligonucleotides, dexamethasone and medium alone (control), respectively. AMs and media were collected after culture for 36 h. The mRNA and protein expression of STAT1 and ICAM-1 in AMs were detected by RT-PCR and Cell-ELISA, respectively. The conditioned media were co-cultured with lung fibroblasts for 30 h, and then the cell proliferation and the concentration of hydroxyproline were examined. RESULTS: (1) The STAT1 mRNA expression by AMs in the STAT1 antisense oligonucleotides group (31.8 +/- 3.5) was lower than those of AMs in the STAT1 sense oligonucleotides group (64.2 +/- 4.3), the dexamethasone group (44.1 +/- 4.6) and the control group (65.5 +/- 4.6) (P < 0.05). Moreover, the STAT1 mRNA expression by AMs in the dexamethasone group was also lower than those of AMs in the STAT1 sense oligonucleotides group and the control group (P < 0.05), but the STAT1 mRNA expression by AMs in the STAT1 sense oligonucleotides group was not different from that of the control group (P > 0.05). The STAT1 mRNA expression by AMs in the NS group (14.9 +/- 3.1) was lower than those of AMs in the STAT1 antisense oligonucleotides group, the STAT1 sense oligonucleotides group, the dexamethasone group and the control group (P < 0.05). (2) The mRNA expression of ICAM-1 showed similar changes to the STAT1 mRNA expression by AMs. (3) The STAT1 protein expression by AMs in the STAT1 antisense oligonucleotides group (4.4 +/- 0.6) or in the NS group (3.7 +/- 0.4) was lower than those of AMs in the STAT1 sense oligonucleotides group (7.7 +/- 0.7), the dexamethasone group (5.9 +/- 0.4) and the control group (7.6 +/- 0.6) (P < 0.05); and the STAT1 protein expression by AMs in the dexamethasone group was also lower than those of AMs in the STAT1 sense oligonucleotides group and the control group (P < 0.05), but the STAT1 protein expression by AMs in the STAT1 sense oligonucleotides group was not different from that of the control group (P > 0.05). (4) The changes of ICAM-1 protein expression, lung fibroblast proliferation and hydroxyproline concentration were consistent with the changes of STAT1 protein expression by AMs. CONCLUSIONS: STAT1 antisense oligonucleotides could inhibit the mRNA and the protein expression of STAT1 and ICAM-1 in AMs. STAT1 antisense oligonucleotides also inhibited lung fibroblast proliferation and hydroxyproline secretion. PMID- 16255960 TI - [Advance in diagnosis and management for thymoma]. PMID- 16255959 TI - [Changes of blood coagulative and fibrinolytic systems and functions of pulmonary vascular endothelium in patients with pulmonary thromboembolism]. AB - OBJECTIVE: To explore the changes of blood coagulative and fibrinolytic systems and functions of pulmonary vascular endothelium in patients with pulmonary thromboembolism (PTE). METHODS: Twenty patients with acute massive PTE, 40 patients with acute non-massive PTE and 40 control subjects without PTE were included in the study. D-Dimer (D-D), tissue-type plasminogen activator (t-PA), plasminogen activator inhibitor1 (PAI-1), plasma protein S (Ps), plasma protein C (Pc), thrombomodulin (TM), anticardiolipin antibody (ACA) and homocysteine (Hcy) were measured by the method of ELISA. Antithrombin-III (AT-III) activity was measured by chromo-substrate method. RESULTS: The levels of D-D, t-PA, PAI-1, and TM were (1.46 +/- 0.62) mg/L, (11.4 +/- 6.9) microg/L, (88.2 +/- 27.5) microg/L, (6.8 +/- 1.1) microg/L respectively in patients with acute massive PTE and (0.92 +/- 0.27) mg/L, (6.6 +/- 1.5) microg/L, (60.1 +/- 26.1) microg/L, and (6.30 +/- 1.50) mg/L in patients with acute non-massive PTE. The levels of both PET groups were significantly higher than those of the control subjects [(0.38 +/- 0.10) mg/L, (4.7 +/- 1.4) microg/L, (35.7 +/- 9.2) microg/L, (3.0 +/- 0.5) microg/L and P < 0.05]. The levels of AT-III were (86.0 +/- 11.8)% in patients with acute massive PTE and (90.1 +/- 9.0)% in patients with acute non-massive PTE. The levels of AT-III in both groups were significantly lower than those of the control subjects, which were (102.6 +/- 9.2)% (P < 0.01 and P < 0.05 respectively). The levels of ACA-IgG, IgM and IgA in patients with acute massive PTE and non-massive PTE were also significantly higher than those in the control group (P < 0.05). CONCLUSION: Imbalance of blood coagulation and fibrolytic systems and pulmonary vascular endothelium damage occur in patients with PTE. PMID- 16255972 TI - [How far away from stem cell transplant for clinic use?]. PMID- 16255961 TI - [Mycotoxins in lung diseases]. PMID- 16255974 TI - [Ethical problems in human stem cell research and application]. PMID- 16255975 TI - [Adult stem cells and their possible application]. PMID- 16255977 TI - [Brightness VS Challenge: adult stem cells application in corneal and ocular surface disease]. PMID- 16255978 TI - [Spontaneous malignant transformation of adult stem cell in vitro culture proliferation]. PMID- 16255980 TI - [Cellular phenotype conversion induced by co-culture of human mesenchymal stem cells cocultured with human sweat gland cells]. AB - OBJECTIVE: To investigate the cellular phenotype conversion during human mesenchymal stem cells (hMSCs) cocultured with injured human sweat gland cells (hSGCs) in vitro. METHODS: HMSCs and hSGCs were isolated and cultured and expanded respectively. The antigens expression of hMSCs and hSGCs were detected by two-steps immunocytochemistry. HMSCs were labeled with BrdU. The hSGCs were heat-shocked at 47 degrees C for 40 min when they reached 70% confluency, then cooled for 1-2 h at 37 degrees C and (1 - 2) x 10(5) BrdU-labeled hMSCs were added before incubation for up to 2 weeks. The cocultures were observed by phase contrast microscopy and detected by double-staining immunocytochemistry using CEA and BrdU as primary antibodies. RESULTS: The cultured hMSCs and hSGCs were clonogenic growth. HMSCs were positive for anti-CD44 and anti-CD105 staining and negative for anti-CD34 and anti-CEA staining. HSGCs express CK7, CK18, CK19 and CEA. The positive rate of BrdU labeled-hMSCs was 90%. The majority of hSGCs lost cell-cell contact after heat-shock. 2 weeks after cocultured, some cocultured cells were positive for both anti-CEA and anti-BrdU staining and some cocultures had more than two nuclei which stained with two different colors by double staining immunocytochemistry. Statistic results showed 1%-5% of the hMSCs added to the coculture system were recovered as double-staining cells expressing BrdU and CEA while only 0.01%-0.05% cells stained with two different colors in nuclei. The multi-nucleated cells were wide and flatten. CONCLUSION: HMSCs could differentiate into hSGCs in vitro under injured microenvironment. The mechanisms of which may be that hMSCs differentiate into hSGCs directly or by cell fusion, even nucleus fusion. PMID- 16255982 TI - [Human adipose derived stem cells suppress lymphocyte proliferation induced by cellular or nonspecific mitogenic stimuli]. AB - OBJECTIVE: To study immunological properties of adipose derived stem cells (ADSC) and their in vitro immunomodulatory effects on lymphocytes. METHODS: ADSC was isolated from fat tissue by liposuction and culture expanded. Cells at passage 2 were observed for the expression of HLAI, HLAII by FACs analysis. Peripheral blood lymphocytes (PBL) were cultured with allogeneic ADSCs at dose of 1 x 10(5). A third-party ADSCs at dose of 1 x 10(2), 1 x 10(3), 1 x 10(4), 1 x 10(5) were added to the ongoing two-way mixed lymphocyte reactions (MLR) for 6 days. Value of CPM (count per minute) was calculated with luminescence counter. RESULTS: Flow cytometry showed that undifferentiated ADSCs express HLA class I but not class II. Addition of interferon gamma (IFN-gamma) for 48 hours induced greater than 90% of cells to express HLA class II. No lymphocyte response was induced by allogeneic ADSCs as stimulators. Results were similar using ADSCs pretreated with IFN-gamma. ADSCs elicited inhibit function of mixed lymphocyte cultures in a dose dependent way. Even if ADSCs were pretreated with IFN-gamma, the suppression ability was maintained. CONCLUSION: Undifferentiated ADSCs do not elicit alloreactive lymphocyte proliferative responses and could modulate immune responses in vitro. PMID- 16255983 TI - [Experimental study of compatibility between chondrocytes and allogenic cartilage microparticle acellular tissue matris in vitro]. AB - OBJECTIVE: To construct tissue engineered cartilage using cartilage microparticle acellular tissue matrix (CMACTM) as scaffold. METHODS: To determine the content of hydroxyproline, glycosaminoglycan and DNA of CMACTM prepared from sheep's articular cartilage with multistep enzymic method, and to analyze CMACTM with gross observation, histology and scanning electron microscopy. Allogenic chondrocytes were mixed with CMACTM and cultured in vitro from 0 to 35 days. Observations through inverted microscope, scanning and transmission electron microscope, quantifications of hydroxyproline, glycosaminoglycan and DNA in the composite, cells adhesion rate were applied to analyze the results. RESULTS: The diameter of CMACTM was 0.100-0.154 mm, which contain extracellular matrix only. Hydroxyproline, glycosaminoglycan and DNA quantifications in CMACTM were 204.374 +/- 3.120 microg/mg, 18.302 +/- 2.037 microg/mg and 0.042 +/- 0.013 microg/mg respectively. Allogenic chondrocytes enclosed CMACTM tightly, hydroxyproline, glycosaminoglycan and DNA quantifications in the composite of the two formers increased with difference on 7th day compared with that on 0 day, reached to the peaks on 14th day (hydroxyproline, DNA) and on 21st day (glycosaminoglycan), and retained at a high level on the following days. Cells adhesion rate was 92%. CONCLUSION: Allogenic CMACTM possessed satisfactory biocompatibility for chondrocytes and provided a new scaffold for cartilage tissue engineering. PMID- 16255984 TI - [Effects of the cutaneous and blood contents of glucose on wound healing in diabetic rats with superficial partial thickness scalding]. AB - OBJECTIVE: To study the effect of the cutaneous and blood contents of glucose on wound healing in diabetic rats with superficial partial thickness scalding. METHODS: 96 Sprague-Dawley rats were randomized into control group and STZ induced diabetic group, inflicted with superficial partial thickness scalding of 10% TBSA on the back. The glucose content in the blood and wound tissue were evaluated before injury and at day 1, day 3, day 5, day 7, day 10 and day 14 after injury. Wound healing process in the rats was dynamically observed by naked eyes and histologically examined. The cell cycles of keratinocytes from wound site were determined before injury and at day 3, day 7 and day 10 after injury. RESULTS: The concentrations of blood glucose in diabetic wound was significant increased than normal group (27.28 mmol/L +/- 0.80 mmol/L vs. 4.65 mmol/L +/- 0.14 mmol/L, P < 0.01). The content of local glucose in wound tissue were significantly correlated with that in the blood (r = 0.881, P < 0.05). When compared with the control group, wound healing of the diabetic rats were delayed with the characteristics of impaired epithelialization and decreased. percentages of S stage and G(2)/M stages of keratinocytes. CONCLUSION: Local glucose content in diabetic wound was varied with blood glucose concentration. The increased glucose concentration in diabetic wound was attributed to the impaired wound healing in diabetes. High glucose concentration could inhibit the epithelialization and decrease the keratinocyte proliferation. PMID- 16255985 TI - [Overexpression of Shp-2 is associated with the unlimited growth and apoptosis resistance of p210 bcr-abl-mediated chronic myeloid leukemia]. AB - OBJECTIVE: To investigate the expression level of Shp-2 tyrosine phosphatase in chronic myeloid leukemia (CML) and its relationship with the unlimited growth and apoptosis resistance of p210 bcr/abl-induced malignant cells. METHODS: In this study, p210 bcr/abl positive leukemia cell specimens were obtained from 25 CML cases, meanwhile, bone marrow and peripheral blood cell samples from 8 non-tumor individuals and 10 normal individuals were used as p210 bcr/abl negative controls. K562 and KU812 leukemia cells were used as p210 bcr/abl positive controls, and KG-1 leukemia cell line was used as Shp-2 positive control. Specimens of peripheral blood and bone marrow of 25 adult patients of chronic myelocytic leukemia, 15 males and 10 females, aged 28-64, were collected. Specimens of bone marrow of 8 basically healthy adult volunteers and specimens of peripheral blood of 10 healthy adult volunteers were used as controls. The total cell protein was collected and the expression of Shp-2 was examined by Western blotting. Human leukemia cells of the line K562 were cultured. Shp-2 specific sense and antisense oligonucleotides were added into the culture fluid respectively. The cell apoptosis was detected by flow cytometry (FCM). STI571, specific inhibitor of p210 bcr/abl was added into the cultured fluid of K562 cells, then Western blotting and FCM were used to detect the protein expression of Shp-2 and p210 bcr/abl, and cell apoptosis. RESULTS: Phosphorylated Shp-2 (pShp)-2 protein was overexpressed in 92% (23/25) of the CML cells, but lowly expressed or not expressed in the normal hematopoietic cells. The mean pShp-2 protein/beta-actin ratio of the primary CML leukemia cells was 0.91 +/- 0.62, significantly higher than those of the normal bone marrow cells and peripheral blood hematopoietic cells (0.16 +/- 0.09 and 0.03 +/- 0.05 respectively, both P < 0.01). The apoptotic rates of the CML cells treated by Shp-2 specific antisense oligonucleotide of the concentrations of 1 micromol/L and 4 micromol/L respectively for 72 h was 7.98% and 20.29% respectively, both significantly higher than that of the control group (4.06%, P < 0.01). The number of clone of CML cells treated by 0.25 micromol/L and 1.0 micromol/L Shp-2 specific antisense oligonucleotide for 7 days were 67% (37/60) and 11.9% (5/42) that of the control group. Twenty-four and 48 hours after the stimulation of STI571 the expression level of Shp-2 protein in the CML cells decreased time-dependently and the CML cell apoptotic rates were 31.15% and 38.69% respectively, both lower than that of the control group (33.6%). The number of clone of CML cells effected by 0.25 micromol/L and 1.0 micromol/L Shp-2 specific antisense oligonucleotide for 7 days was 67% (37/60) and 11.9% (5/42) that of the control group. Twenty-four and 48 hours after the stimulation of STI571 the expression level of Shp-2 protein in the CML cells decreased time-dependently and the CML cell apoptotic rates were 31.15% and 38.69% respectively, both lower than that of the control group (33.6%). CONCLUSION: (1) The pShp-2 protein is overexpressed in CML cells, which is associated with the unlimited growth and apoptosis resistance of malignant cells. (2) Shp-2 is upregulated by p210 bcr/abl oncoprotein in CML. PMID- 16255987 TI - [Effects of different drugs on the sphincter of Oddi motility: study with choledochoscope manometry]. AB - OBJECTIVE: This study assessed the effects of narcotic analgesics, nitrates drugs, somatostatin analogues on human sphincter of Oddi motility and the antagonistic effects of nitrates drugs and anticholinergic agents against morphine measured by choledochoscope manometry. METHODS: 157 patients who had a T tube after cholecystectomy and choledochotomy were assessed by choledochoscope manometry. They were randomly divided into 5 groups. Sphincter of Oddi basal pressure (SOBP), amplitude (SOCA), frequency of contractions (SOF), duration of contractions (SOD), duodenal pressure (DP), common bile duct pressure (CBDP) were scored and analyzed. RESULTS: SOBP, SOCA and SOF increased after injection of morphine and Ap-237, CBDP increased after intramuscular administered morphine. No apparent change occurred after intramuscularly administered pethidine. SOBP and SOCA decreased after tramadol. After intravenous administration of stilamin in a dose of 250 microg/h, SOCA increased. After administration stilamin of 500 microg/h dose, SOCA and SOBP declined. After intravenous administration of sandostatin, CBDP increased obviously. BPOS and SOCA decreased significantly after administration of isosorbide dinitrate (ISDN) and glyceryl trinitrate (GTN), SOBP reduced evidently after application of pentaerythritol tetranitrate (PTN). SOBP, SOCA, SOF and CBDP increased evidently after injection of morphine. After associated application of ISDN and GTN, the four indications above decreased. As to associated application with PTN, SOCA and SOF decreased. After associated application of anisodamine or atropine, SOCA, SOBP declined, after injected buscopan, SOCA, SOBP, SOF all declined. CONCLUSION: The regular dose of morphine and Ap-237 shows excitatory effect on the sphincter of Oddi motility. Tramadol shows inhibitory effect on the sphincter of Oddi. The regular dose of pethidine and sandostatin shows no apparent effect on the sphincter of Oddi. Stilamin in low dose shows excitated effect on the sphincter of Oddi. Nitrates drugs show inhibited effects on SO motility. Nitrates drugs and anticholinergic agents can antagonize the excitated effect of morphine. PMID- 16255986 TI - [A study on the mechanism of islet cell insulin resistance in high-fat-diet obese rats]. AB - OBJECTIVE: To study the changes of insulin signal transduction in islet cells of high-fat-diet rats with peripheral insulin resistance (IR). METHODS: Thirty male Wistar rats were randomly divided into 2 equal groups: high-fat-diet group and control group to be fed with high-fat food and normal food respectively. Twenty weeks after the rats were sacrificed. The contents of insulin and glucagon in homogenate of pancreas were detected during islet cell perifusion, and insulin receptor (IRc) and insulin receptor substrates (IRS-1 and IRS-2) were detected by immunohistochemistry. RESULTS: (1) The insulin sensitive index (ISI) was significantly decreased in the high-fat-diet rats in comparison with the normal rats, while the contents of glucagon in blood and in homogenate of pancreas were both significantly increased in the high-fat-diet rats (362 pg/ml +/- 58 pg/ml vs 291 pg/ml +/- 35 pg/ml; 442 pg/ml +/- 56 pg/ml vs 287 pg/ml +/- 48 pg/ml, both P < 0.05). (2) The glucose stimulated insulin secretion (GSIS) was impaired in the high-fat-diet rats. 16.7 nmol/L glucose could inhibit the glucagon secretion by the alpha cells of the normal rats, but not of the high-fat-diet rats. (3) The expression of IRc, IRS-1 and IRS-2 in islets was stronger in the peripheral cells (non-insulin secretion cells) than in the center cells (insulin secretion cells). The expression of IRc and IRS-2 was significantly decreased by 28% and 22% respectively in the high-fat-diet rats compared with the normal controls (both P < 0.01). CONCLUSION: High-fat-diet rats have impairment of insulin signal transduction in islet cells, which may contribute to the insulin resistance of islet alpha and beta cells and explain, at least in part, the dysfunction of the islet cells under peripheral IR. PMID- 16255988 TI - [Roles of cyclooxygenase-2 and caspase-3 expression in pathogenesis of lung carcinoma: an experiment with rats]. AB - OBJECTIVE: To investigate the dynamic expression of cyclooxygenase-2 (COX-2) and caspase-3 during the carcinogenesis, invasion and metastasis of 3 methylcholanthrene (MCA) and diethylnitrosamine (DEN)-induced rat lung cancer and its significance. METHODS: Iodized oil with MCA and DEN was instilled into the left bronchi of 80 Wistar rats to induce squamous cell lung carcinoma. Iodized oil without MCA and DEN was instilled into the left bronchi of 10 rats as control group. Sixteen rats in the experimental group and 2 rats in the control group were killed 10, 15, 35, 60, and 270 days after experiment respectively to undergo pathological examination. Immunohistochemistry was used to detect the protein expression of COX-2 and caspase-3 in the bronchial endothelial cells. Immunohistochemical scores (IHS) were calculated. RESULTS: Pathological changes of different phases, such as hyperplasia of bronchial mucosal endothelial cells (14 cases), squamous metaplasia (25 cases), atypical proliferation (35 cases), carcinoma in situ (12 cases), infiltrative carcinoma (54 cases), and metastatic carcinoma (15 cases) appeared successively in the left lung mucosal endothelium of the experimental group. Weak expression of COX-2 protein was occasionally seen in the normal bronchial mucosal endothelium. COX-2 protein expression was becoming stronger and the IHS was becoming higher along with the development of carcinoma (P < 0.01). Caspase-2 protein expression was positive in 8 of the 10 (80%) of the control rats with an HIS of 5.92 +/- 0.9. And caspase-2 protein expression was becoming weaker along with the development of carcinoma (P < 0.01 or P < 0.05). Significant negative correlation was found between the COX-2 protein expression and caspase-3 expression (P < 0.01). CONCLUSION: COX-2 and caspase-3 play important roles in the carcinogenesis of MCA and DEN-induced rat lung squamous cell carcinoma. COX-2 may take part in blocking cell apoptosis by inhibiting caspase-3 activity, thereby promoting the carcinogenesis of lung cancer. The imbalance between COX-2 and caspase-3 may be a critical factor affecting the biologic behavior of lung carcinogenesis, invasion and metastasis. PMID- 16255990 TI - [Abnormal expression of Thy-1 as a novel tumor marker in lung cancer and its prognostic significance]. AB - OBJECTIVE: The purpose of this study was to investigate the expression of Thy-1 immunohistochemically in different lung tumors and its prognostic significance in non-small cell lung cancer (NSCLC) cases. We also evaluate relationship between Thy-1 and p53 expression status so as to find any clue about the mechanism. METHODS: In this study, we used anti-Thy-1/CD90 antibody to detect the expression pattern of Thy-1 in different lung tumor sections, which were embedded in paraffin blocks. The expressions of Thy-1 in 175 lung tissue cases, including different pathological types, were analyzed as tissue array form. We also detect expression status in 91 NSCLC among these cases and analyze the relationship between Thy-1 and p53. The relationship between Thy-1 expression and patients' survival was studied. RESULTS: We first found that anti-Thy-1 antibody can strongly stain a nuclear molecule in different type of lung cancer cells. Among lung cancer cases, 89 (56.7%) cases showed strong nuclear staining for Thy-1 specially. In univariate and multivariate analysis for 91 NSCLC patients we found TNM staging, lymph node status and Thy-1 overexpression in nuclei were independent factors to affect the prognosis of NSCLC patients. In lymph node non metastasis subgroup cases, Thy-1 negative patients had significant longer survival than Thy-1 positive cases (mean survival: 46.42 mons vs 38.56 mons, P = 0.0207). There was a significant association between Thy-1 and p53 expression (P < 0.0001). CONCLUSION: There is a significant overexpressed Thy-1 located in lung cancer cell nucleus as compared to the normal tissue or benign tumor cells of lung, and it is one of the factors effected on the prognosis of NSCLC patients. This finding suggests that Thy-1 maybe a novel latent malignant marker in the lung cancer pathology. The association between Thy-1 and p53 expression in nucleus suggests that p53 protein and Thy-1 may have some interaction. PMID- 16255991 TI - [Study of pharmacokinetics/pharmacodynamics of levofloxacin]. AB - OBJECTIVE: To evaluate the dosing regimens of levofloxacin. METHODS: The drug concentrations in serum and urine were assayed by HPLC method and the pharmacokinetic parameters were calculated after intravenous infusion of a single dose of 200 mg, 300 mg and 500 mg levofloxacin to healthy volunteers. The in vitro activity MIC of levofloxacin against 823 clinical isolates were determined and compared with other antimicrobial agents. Based on the above results, the PK/PD parameters C(max)/MIC and AUC/MIC were calculated and the dosing regimens of levofloxacin were proposed for infections caused by different pathogens. RESULTS: The results of clinical pharmacokinetic study showed that the C(max) of levofloxacin was 3.4 mg/L +/- 0.8 mg/L, 4.8 mg/L +/- 1.4 mg/L and 7.6 mg/L +/- 1.1 mg/L respectively, AUC(0-infinity) was 14.4 mg.h/L +/- 2.5 mg.h/L, 21.9 mg.h/L +/- 4.5 mg.h/L and 38.3 mg.h/L +/- 4.9 mg.h/L respectively, T(1)/2 beta was 6.2 h +/- 0.4 h, 6.4 h +/- 0.9 h and 6.5 h +/- 0.6 h respectively, and 69% +/ 5%, 69% +/- 6%, and 65% +/- 4% of the doses were excreted in urine within 24 h after intravenous infusion of a single dose of levofloxacin 200 mg, 300 mg and 500 mg. The in vitro pharmacodynamic study showed that levofloxacin was highly active against Hemolytic streptococcus and Moraxella catarrhalis. It was active against Streptococcus pneumoniae (including penicillin nonsusceptible strains), Hemophilus influenzae, methicillin-sensitive Staphylococcus aureus (MSSA), Klebsiella pneumoniae and Stenotrophomonas maltophilia. Levofloxacin also had good activity against Pseudomonas aeruginosa and K.pneumoniae. However, most of isolates of enterococcal spp. were resistant to levofloxacin. Above 50% of Escherichia coli isolates were resistant to levofloxacin. Based on the results of PK/PD parameters, the adequate dosing regimens of levofloxacin should be once daily 200 mg once daily of levofloxacin was expected to be effective for the treatment of infections caused by M. catarrhalis. The regimen of 300 mg once daily could be effective for the treatment of infections caused by Hemolytic streptococcus. 500 mg once daily of levofloxacin was expected to be effective for the treatment of respiratory tract infections caused by S. pneumoniae or H. influenzae. For treatment of respiratory tract infections and urinary tract infections caused by levofloxacin-susceptible organisms including K. pneumoniae, E. coli, P. aeruginosa, and S.maltophilia, 500 mg once daily of levofloxacin was needed to obtain good clinical efficacy. But PK/PD parameters predicted that 500 mg daily of levofloxacin was not effective for infections caused by methicillin resistant Staphylococcus aureus (MRSA) and Enterococci. CONCLUSION: The proposed dosing regimens of levofloxacin based on PK/PD concepts are expected to provide good efficacy in clinical practice. PMID- 16255993 TI - [Triple-type theory of statistics and its application in the scientific research of biomedicine]. AB - OBJECTIVE: To point out the crux of why so many people failed to grasp statistics and to bring forth a "triple-type theory of statistics" to solve the problem in a creative way. METHODS: Based on the experience in long-time teaching and research in statistics, the "three-type theory" was raised and clarified. Examples were provided to demonstrate that the 3 types, i.e., expressive type, prototype and the standardized type are the essentials for people to apply statistics rationally both in theory and practice, and moreover, it is demonstrated by some instances that the "three types" are correlated with each other. It can help people to see the essence by interpreting and analyzing the problems of experimental designs and statistical analyses in medical research work. RESULTS: Investigations reveal that for some questions, the three types are mutually identical; for some questions, the prototype is their standardized type; however, for some others, the three types are distinct from each other. It has been shown that in some multifactor experimental researches, it leads to the nonexistence of the standardized type corresponding to the prototype at all, because some researchers have committed the mistake of "incomplete control" in setting experimental groups. This is a problem which should be solved by the concept and method of "division". CONCLUSION: Once the "triple-type" for each question is clarified, a proper experimental design and statistical method can be carried out easily. "Triple-type theory of statistics" can help people to avoid committing statistical mistakes or at least to decrease the misuse rate dramatically and improve the quality, level and speed of biomedical research during the process of applying statistics. It can also help people to improve the quality of statistical textbooks and the teaching effect of statistics and it has demonstrated how to advance biomedical statistics. PMID- 16255996 TI - Vascular abnormalities in the metabolic syndrome: mechanisms and therapy. PMID- 16255997 TI - Undesirable effects of extreme dietary carbohydrate and saturated fat intakes: the search for the middle ground. PMID- 16255998 TI - Effect of very high-fat diets on body weight, lipoproteins, and glycemic status in the obese. AB - Given the increased prevalence of obesity in the United States, despite reduced fat intake, there has been increasing interest in the effect of dietary fat on body weight, lipoproteins, and glycemic status. Despite predictions from epidemiologic and physiologic studies, recent prospective trials have demonstrated equivalent weight loss on high-fat versus low-fat diets. Nevertheless, the type of dietary fat consumed has substantially different effects on lipoproteins. Saturated fat raises high-density lipoprotein cholesterol but has unfavorable effects on total cholesterol, and has been associated with increased cardiovascular events. In contrast, unsaturated fats, and particularly omega-3 fatty acids, have the combined benefits of lowering serum cholesterol and raising high-density lipoprotein, as well as favorable effects on insulin resistance and inflammation; they also lower cardiovascular events in high-risk patients. Although current national guidelines modestly liberalize unsaturated fat consumption, important questions still remain about the optimal percentage of unsaturated fats in the diet. PMID- 16255999 TI - The effects of low-fat, high-carbohydrate diets on plasma lipoproteins, weight loss, and heart disease risk reduction. AB - Although there is consensus about restriction of dietary saturated and trans fatty acids, cholesterol, and sugars, there is debate about what the optimal total fat and carbohydrate content of the diet should be for weight loss and coronary heart disease (CHD) risk reduction. The overall evidence that dietary composition plays an important role in determining caloric intake is limited. Three recent randomized trials have indicated that low-carbohydrate diets are more effective in promoting weight loss in overweight and obese subjects over 4 to 6 months, but not over 1 year. In our own randomized trial no such differences were noted, and compliance with extreme diets was limited. Moreover little attempt has been made to control for the type of carbohydrate used in the low fat, high-carbohydrate arms of these trials. Available evidence suggests that restriction of sugars and carbohydrates having a high glycemic index would be preferable to total carbohydrate restriction, and that an increased intake of fiber and essential fats (especially omega-3 fatty acids) is also important for overall heart disease risk reduction. PMID- 16256000 TI - Benefits and hazards of dietary carbohydrate. AB - Since the dawn of civilization, carbohydrate has comprised the largest source of energy in the diet for most populations. The source of the carbohydrate has been from plants in the form of complex carbohydrate high in fiber. Only in affluent cultures has sugar contributed so much of the total energy. When carbohydrate is consumed as a major component of a plant-based diet, a high-carbohydrate, low-fat diet is associated with low plasma levels of total and low-density lipoprotein cholesterol, less coronary heart disease, less diabetes, and less obesity. Very low-carbohydrate (ketogenic) diets may provide short-term solutions but do not lead to a long-term solution for most people. PMID- 16256001 TI - Effects of dietary fats versus carbohydrates on coronary heart disease: a review of the evidence. AB - Recommendations arising from the traditional diet-coronary heart disease (CHD) paradigm, which focuses on effects of total and saturated fat on serum total and low-density lipoprotein cholesterol, may have failed to reduce CHD risk and inadvertently exacerbated dyslipidemia, insulin resistance, and weight gain, particularly among individuals who are older, female, sedentary, or obese. A suitable dietary paradigm must consider types and qualities of fats and carbohydrates consumed, their effects on a range of intermediary risk factors, and characteristics that may modify individual susceptibility. Based on current evidence, replacement of total, unsaturated, and even possibly saturated fats with refined, high-glycemic index carbohydrates is unlikely to reduce CHD risk and may increase risk in persons predisposed to insulin resistance. In contrast, a diet that is 1) rich in whole grains and other minimally processed carbohydrates; 2) includes moderate amounts of fats (approximately 30%-40% of total energy), particularly unsaturated fats and omega-3 polyunsaturated fats from seafood and plant sources; 3) is lower in refined grains and carbohydrates; and 4) eliminates packaged foods, baked goods, and fast foods containing trans fatty acids, will likely reduce the risk of CHD. PMID- 16256002 TI - Influence of the DASH diet and other low-fat, high-carbohydrate diets on blood pressure. AB - The Dietary Approaches to Stop Hypertension (DASH) and DASH-sodium trials were controlled feeding dietary trials that lowered blood pressure in the absence of weight loss. The beneficial aspect of DASH appears to be the low saturated fat content (< 7%). Sodium restriction added additional blood pressure lowering to the low saturated fat DASH diet. Sodium restriction was more effective with increasing age and more effective than increasing fruit and vegetable content. When achievement of sodium restriction, exercise, and weight loss goals were reached in the outpatient setting with subjects making their own food choices (as in the PREMIER study), adding the DASH diet with an average fruit and vegetable intake of 7.8 servings daily had no additional benefit in those younger than 50 years of age or in ethnic/gender subgroups, but did have a benefit for the total group older than age 50 years. Because many hypertensive subjects are overweight, hypocaloric versions of DASH geared toward weight loss are appropriate. Mechanisms for dietary beneficial effects are related to inflammation and insulin sensitivity. PMID- 16256003 TI - Influence of dietary carbohydrate and fat on LDL and HDL particle distributions. AB - Variations in the size and density distributions of low-density lipoprotein (LDL) and high-density lipoprotein (HDL) particles have been related to risk for cardiovascular disease. In particular, increased levels of small, dense LDL particles, together with reduced levels of large HDL and increases in small HDL, are integral features of the atherogenic dyslipidemia found in patients with insulin resistance, obesity, and metabolic syndrome. Increased dietary carbohydrates, particularly simple sugars and starches with high glycemic index, can increase levels of small, dense LDL and HDL, primarily by mechanisms that involve increasing plasma triglyceride concentrations. Low-carbohydrate diets may have the opposite effects. Diets with differing fatty acid composition can also influence LDL and HDL particle distributions. PMID- 16256004 TI - The response of lipoproteins to dietary fat and cholesterol in lean and obese persons. AB - Individuals differ in the response of their blood lipoproteins to cholesterol lowering diets. One characteristic clearly associated with susceptibility to diet is leanness; many studies show that total cholesterol, low-density lipoprotein cholesterol, and high-density lipoprotein cholesterol concentrations respond more strongly to dietary fat and cholesterol in lean subjects than in obese subjects. This is unlikely to be due to differences in dietary compliance. A metabolic explanation is that obese people have a higher rate of total body cholesterol synthesis. The low-density lipoprotein receptors in their liver cells are partly suppressed by this large stream of endogenous cholesterol coming in from their enterohepatic circulation, and the amount added by dietary cholesterol relative to the endogenous pool would be less than in lean people. Whatever the mechanism, diets low in saturated fat and cholesterol are less effective in the obese. The most effective way for obese people to normalize their blood lipids is to lose weight, which is, unfortunately, hard to do in our society. PMID- 16256005 TI - Cholesterol absorption: influence of body weight and the role of plant sterols. AB - Cholesterol absorption and synthesis are inter-regulated, so if one changes then the other changes in the opposite direction. The regulation and detailed mechanism of cholesterol absorption have been intensely investigated. Inhibition of cholesterol absorption has become an additional factor for cholesterol lowering. Agents inhibiting cholesterol absorption, mainly plant stanols and sterols or ezetimibe, usually lower low-density lipoprotein cholesterol in monotherapy by less than 20%, indicating that these inhibitors can normalize cholesterol levels only in patients with a modest baseline hypercholesterolemia. Body weight, especially obesity with and without diabetes, and dietary plant sterols alter cholesterol absorption differently. This article briefly reviews some special questions of cholesterol absorption under these conditions. PMID- 16256006 TI - Gender differences in lipoprotein metabolism and dietary response: basis in hormonal differences and implications for cardiovascular disease. AB - The transport of fat in the blood stream is approximately twice as fast in women as men. Disease states such as obesity and diabetes are associated with greater lipoprotein abnormalities in women compared with men. A greater increment in cardiovascular disease risk in women is linked to these abnormalities. A greater change in triglyceride level and a lesser change in low-density lipoprotein are observed in women than men with high-carbohydrate or high-fat feeding. Most consistent are greater changes in high-density lipoprotein (HDL), HDL2, and apolipoprotein A-I levels in women compared with men with high-carbohydrate or high-fat feeding. Dietary fat restriction in women appears to have a less beneficial lipoprotein effect than in men. Dietary fat restriction for heart disease prevention may be less ideal in women than in men. PMID- 16256007 TI - Statins and aortic stenosis progression: are biologic targets still an option? PMID- 16256008 TI - An update on renovascular hypertension. AB - Renovascular hypertension (RVH) represents a secondary and potentially remediable form of hypertension. Elevated blood pressure is only one of a broad array of pathophysiologic consequences that are associated with decreased renal perfusion. Our ability to accurately and noninvasively detect stenotic lesions within the renal artery is growing. However, functional assessment of renal parenchyma and hemodynamic significance of renal artery lesions is still limited. Advances in endovascular techniques spurred interest in the concept of ischemic nephropathy and the effect of renal artery revascularization on renal function. Despite the relative frequency of atherosclerotic renal artery stenosis (ARAS), there currently is no consensus on the most appropriate therapy. In this article, we focus on the two most common causes of RVH, ARAS and fibromuscular dysplasia. We discuss the therapeutic strategies, disease mechanisms, clinical findings, evolving trends, and developments. PMID- 16256009 TI - Primary aldosteronism: diagnostic and therapeutic considerations. AB - Recent evaluations indicate that primary aldosteronism (PA) is common in patients with hypertension. In patients with mild to moderate hypertension the prevalence of PA is 5% to 10%, whereas in subjects with resistant hypertension the prevalence is approximately 20%. As such, PA has become the most common secondary cause of hypertension. Such high prevalence rates are distinctly different from earlier assessments in which PA was found to be rare, with a prevalence of generally less than 1% of hypertensive patients. Why PA is seemingly so much more common now than when first described remains unknown. Accurate identification of PA allows for specific therapy with aldosterone antagonists or with surgical resection of aldosterone-producing adenomas. Determination of the plasma aldosterone to plasma renin activity ratio is an effective screen for PA in that it has a high negative predictive value even in the setting of ongoing antihypertensive therapy. Its specificity, however, is low such that a high ratio is suggestive of PA but must be confirmed by demonstration of high and autonomous secretion of aldosterone. PMID- 16256010 TI - Miscellaneous endocrine causes of hypertension. AB - Aldosterone is the principal circulating mineralocorticoid in humans, and aldosterone synthesis normally occurs in the face of volume depletion and renin stimulation. In primary and secondary aldosteronism, aldosterone synthesis continues despite volume expansion and causes hypertension. Other steroid hormones that are aldosterone and cortisol precursors also activate the mineralocorticoid receptor and cause hypertension when overproduced. Mineralocorticoid synthesis in these pathologic states can be constitutive or driven by pituitary adrenocorticotropic hormone (ACTH), due to genetic defects that cause disordered steroid synthesis or catabolism. This review focuses on uncommon forms of ACTH-dependent mineralocorticoid excess states associated with hypertension. PMID- 16256012 TI - Obstructive sleep apnea and hypertension. AB - There is growing evidence of a causal relationship between obstructive sleep apnea (OSA) and hypertension. Untreated OSA may have direct and deleterious effects on cardiovascular function and structure through several mechanisms, including sympathetic activation, oxidative stress, inflammation, and endothelial dysfunction. OSA may contribute to or augment elevated blood pressure levels in a large proportion of the hypertensive patient population. It is important to consider OSA in the differential diagnosis of hypertensive patients who are obese. OSA should be especially considered in those hypertensive patients who respond poorly to combination therapy with antihypertensive medications. PMID- 16256011 TI - Coarctation of the aorta. AB - Coarctation of the aorta is an important, treatable cause of secondary hypertension. Its prevalence varies from 5% to 8% of all congenital heart defects. This condition is most often detected because of a murmur or hypertension found on routine examination. Delayed or absent femoral pulses and an arm/leg systolic blood pressure difference of 20 mm Hg or more in favor of the arms may be considered as evidence for aortic coarctation. The coarctation may be demonstrated on a suprasternal notch two-dimensional echocardiographic view along with increased Doppler flow velocities across the coarctation site. Cardiac catheterization reveals significant systolic pressure gradient (> 20 mm Hg) across the coarctation and angiography demonstrates the degree and type of aortic narrowing. Aortic obstruction may be relieved by surgery or by transcatheter techniques; the latter include balloon angioplasty and stent implantation. In the past, surgery has been used exclusively, but because of morbidity and complications associated with surgery, catheter techniques are increasingly used in the management of aortic coarctation. Balloon angioplasty in children and stents in adolescents and adults are becoming initial therapeutic options for management of coarctation. Studies evaluating long-term follow-up results of the interventional techniques are needed. PMID- 16256016 TI - Lipid therapy for cardiovascular disease with insulin resistance, diabetes, or the metabolic syndrome. AB - Insulin resistance with the development of diabetes or metabolic syndrome is associated with characteristic lipid abnormalities and an increased likelihood of cardiovascular events. This review examines the rationale and clinical trial evidence for the benefit of lipid drug therapy in the presence of these related high-risk conditions. In particular, the results of the more recent statin trials are discussed and contrasted with evidence from trials with fibrates, which, although fewer in number, appear to show that this particular therapy has selective benefit for the overweight individual with diabetes or features of the metabolic syndrome. PMID- 16256015 TI - Treatment of dyslipidemia in children and adolescents. AB - The early lesions of atherosclerosis begin in childhood, and are related to antecedent cardiovascular disease risk factors. Environmental and genetic factors such as diet, obesity, exercise, and certain inherited dyslipidemias influence the progression of such lesions. The identification of youth at risk for atherosclerosis includes an integrated assessment of these predisposing factors. Treatment starts with a diet low in total and saturated fat and cholesterol, the use of water-soluble fiber and plant sterols, weight control, and exercise. Drug therapy, for example, with inhibitors of hydroxymethylglutaryl CoA reductase, bile acid sequestrants, and cholesterol absorption inhibitors, can be considered in those with a positive family history of premature coronary artery disease and a low-density lipoprotein cholesterol above 160 mg/dL, after dietary and hygienic measures. Candidates for drug therapy often include those with familial hypercholesterolemia, familial combined hyperlipidemia, the metabolic syndrome, polycystic ovarian syndrome, type I diabetes, and the nephrotic syndrome. We review the safety and efficacy of dietary and drug therapy, and propose an updated diagnostic and therapeutic algorithm that includes the metabolic syndrome. The early identification and treatment of youth with dyslipidemias is likely to retard the atherosclerotic process. PMID- 16256017 TI - Obesity and lipids. AB - Obesity increases cardiovascular risk through multiple mechanisms. Abdominal (visceral) adiposity is metabolically active and is largely responsible for the atherogenic dyslipidemia, hyperinsulinemia, hypertension, chronic inflammatory state, and prothrombotic state that constitute the metabolic syndrome, and the subsequent increased risk for cardiovascular disease and acute coronary events. Cholesterol guidelines for assessing cardiovascular risk have traditionally focused on low-density lipoprotein (LDL) levels, and reduction of plasma LDL has been shown to reduce cardiovascular events and total mortality. However, the cardiovascular risks associated with the dyslipidemia of obesity--characterized by low levels of high-density lipoprotein; increased triglycerides; increased subfractions of small, dense LDL; and increased levels of apolipoprotein B-100- are also now well recognized. PMID- 16256018 TI - Effective use of combination lipid therapy. AB - Despite the benefits of statin therapy, low-density lipoprotein cholesterol (LDL C) management remains suboptimal and many patients do not achieve their recommended target goals. The aim of combination lipid drug therapy in high-risk patients is to achieve LDL-C and non-high-density lipoprotein cholesterol (HDL-C) goals with a minimum of serious adverse effects. Although statins are the drug of first choice, statin monotherapy may be limited by intolerance of dose escalation or failure to attain non-HDL-C goals in those with mixed hyperlipidemia. Statins plus bile acid resins or ezetimibe can achieve greater than 50% reduction in LDL C, with little or no increase in adverse effects. Fibrates, niacin, and omega-3 fatty acids, when added to statins, can reduce triglycerides, increase HDL-C, and reduce non-HDL-C to a greater extent than statin monotherapy. The safety profile of combination lipid therapy is acceptable, if the global coronary heart disease risk of the patient is high, thus producing a favorable risk to benefit ratio. Careful surveillance of hepatic transaminases, avoidance of gemfibrozil in statin fibrate combinations, and awareness of statin-concomitant drug interactions is key to safe and efficacious use of combination lipid drug therapy. PMID- 16256020 TI - [Construction and identification of ureB gene vaccine of H. pylori]. AB - AIM: To clone ureB gene of H.pylori and construct its gene vaccine. METHODS: ureB gene was amplified by PCR from genome of H. pylori 11637 strain and subcloned into pMD18-T vector. The vector digested with restriction enzyme (Sal I and Bgl II) was inserted into pTCAE and transformed into E. coli DH5alpha. The positive recombinant plasmid identified by digesting with restriction enzyme (Sal I and Xho I) and sequencing named pT-ureB. The pT-ureB was transfected into CHO cells by electroporation method. The expression of UreB protein was detected by Western blot. RESULTS: The pT-ureB was obtained by cloning and recombinant DNA technique. The Western blot analysis showed that the expression of UreB protein (M(r)approximately 62,000) was detected in culture supernatant of CHO cells following transfection with pT-ureB. CONCLUSION: UreB DNA vaccine of H. pylori was successfully constructed. The expression of UreB protein can be detected in culture supernatants of transfected CHO cells. PMID- 16256019 TI - Genetic variation and lipid metabolism: modulation by dietary factors. AB - Cardiovascular diseases (CVD) result from complex interactions between genetic and environmental factors. The evidence supports that gene-environment interactions modulate plasma lipid concentrations and potentially CVD risk. The findings from studies examining gene-diet interactions and lipid metabolism have been promising. Several loci (eg, APOA1, APOE, LIPC) are providing proof of concept for the application of genetics in the context of personalized nutrition for CVD prevention. The spectrum of candidate genes has been expanding to incorporate those involved in intracellular lipid metabolism (eg, iPPARs, CYP7A1). However, the practical application of these findings is not ready for prime time. There is a compelling need for replication using a higher level of scientific evidence. Moreover, we need to evolve from the simple scenarios examined nowadays (ie, one single dietary component, SNP, and risk factor) to more realistic situations involving multiple interactions. In summary, there is need for both large population studies and well-standardized intervention studies. PMID- 16256021 TI - [Construction of recombinant adenovirus expressing sCD40L-Ig]. AB - AIM: To construct adenovirus expressing secretory human CD40L-Ig (sCD40 ligand Ig fusion protein). METHODS: The genes encoding the extracellular domain of human CD40L and IgG Fc were amplified with PCR and inserted into shuttle vector pAdTrack-CMV which then was transformed into E. coli pAdEasy-1-BJ5183 to produce recombinant Ad plasmid. The recombinant plasmid was digested with Pac I and transfected into 293 cells to generate recombinant adenovirus. The recombinant adenovirus was then used in mixed lymphocyte reaction (MLR) to test its function. RESULTS: The recombinant adenovirus sCD40L-Ig was produced. It could inhibit the lymphocyte proliferation in MLR. CONCLUSION: The sCD40L-Ig adenovirus is prepared successfully and its inhibition of MLR is confirmed. PMID- 16256022 TI - [Activity analysis of human beta beta2-microglobulin gene promoter in P815 cells]. AB - AIM: To clone the human beta2-microglobulin(beta2m) gene promoter and study its activity in P815 cells. METHODS: PCR amplification was performed using primers based on human beta2m gene sequence from GenBank and human genomic DNA as a template. The PCR product was directedly ligated into pBluescript II vector for sequencing. The promoter fragment was subcloned into a pcDNA3-EGFP plasmid after it had been identified correctly. A mouse mastocytoma cell line P815 was transiently and stably transfected with the plasmid containing human beta2m gene promoter and enhanced green fluorescence protein(EGFP) gene. The mRNA expression of EGFP in transiently transfected cells was quantified by RT-PCR and that in stably transfected cells was detected by fluorescence microscope and flow cytometry(FCM). RESULTS: A 302 bp DNA fragment was amplified and cloned into the pcDNA3-EGFP vector. RT-PCR analysis showed that EGFP mRNA expression was induced by IFN-gamma in a dose-dependent manner. There was no difference in the fluorescence positive cell rate between the IFN-gamma-treated group and the control group. But the fluorescence intensity of the 5 x 10(5) U/L IFN-gamma treated group increased about 2 folds compared with that of the control group. CONCLUSION: Human beta2m gene promoter is active in mouse mastocytoma P815 cells. It can regulate the expression of reporter gene under the control of IFN-gamma. PMID- 16256023 TI - [HLA-G1 molecule expressed by ECV304 cells inhibit cytotoxic activity of allogeneic NK cells]. AB - AIM: To study the effect of HLA-G1 molecule expressed by an endothelial cell line (ECV304) on the cytotoxic activity of allogeneic NK cells. METHODS: ECV304 cells were transfected with recombinant plasmid pcDNA3-HLA-G1 by the liposome transfection, and the expressed HLA-G1 on the cell surface was detected by indirect immunofluorescent assay and flow cytometry. The cytotoxic activity of allogeneic NK cells against ECV304 cells was analyzed by the MTT method. RESULTS: HLA-G1 was expressed on the surface of the transfected ECV304 cells. The specific lysis of NK cells against plasmid pcDNA3 transfected ECV304 was (50.6+/-18.1)%, while the specific lysis against pcDNA3-HLA-G1 transfected ECV304 was (29.7+/ 11.4)%, which was significantly lower than the former (P<0.001). CONCLUSION: HLA G1 expressed by the ECV304 cells can inhibit cytotoxicity of allogeneic NK cells. PMID- 16256024 TI - [Effects of TGFbeta1 on the transcriptional activity of SP1, AP1 and Smad3-Smad4 in lung fibroblasts]. AB - AIM: To investigate the effects of TGFbeta1 on the transcriptive activity of SP1, AP1 and Smad3-Smad4 in rat lung fibroblasts(RLF). METHODS: RLFs were isolated and cultured and then treated with 10 microg/L TGFbeta1. The transcriptive activity of SP1, AP1 and Smad3-Smad4 and the expression of plasminogen activator inhibitor I(PAI-1) and collagen I were detected by means of electrophoretic mobility shift assay (EMSA), Western blot and immunohistochemical staining. RESULTS: After treatment with 10 microg/L TGFbeta1, AP1, SP1 and Smad3-Smad4 were activated and the expression of collagen I and PAI-1 were increased. CONCLUSION: TGFbeta1 may activate SP1, AP1 and Smad3-Smad4 and thus increase the expression of PAI-1 and collagen I. PMID- 16256025 TI - [Expression of IgG receptor on human umbilical vein endothelial cells stimulated with TNF-alpha and IFN-gamma]. AB - AIM: To detect the expression of IgG receptors (FcgammaR) on cytokine-stimulated human umbilical vein endothelial cells (HUVECs). METHODS: By using ELISA, immunocytochemical staining, immunofluorescent staining and RT-PCR, the expression and subtypes of FcgammaR were detected. RESULTS: Non-stimulated HUVECs expressed very low level of FcgammaRIIa. FcgammaRIIa mRNA was dramatically up regulated upon 24 hour stimulation with tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma(IFN-gamma). ELISA results indicated that the expression of FcgammaRIIa increased 16 folds after stimulation with TNF-alpha and IFN-gamma for 3 days (P<0.01). Immunofluorescent staining showed that FcgammaRIIa was expressed on the surface of the stimulated HUVECs. CONCLUSION: TNF-alpha and IFN-gamma could increase FcgammaRIIa expression on HUVECs. The enhanced expression of FcgammaRIIa may mediate the deposition of immune complexes to blood vessels under vasculitic conditions. PMID- 16256026 TI - [The expression of TNF-alpha in the intestine of mice irradiated by neutron and gamma rays]. AB - AIM: To study the expression of tumor necrosis factor alpha (TNF-alpha) in the intestine of mice irradiated by neutron and gamma rays. METHODS: 350 male BALB/c mice were irradiated with neutron and gamma rays of different doses, and sacrificed at 6 and 12 hours, 1, 2, 3, 4, 5, 7, 10, 14, 21 and 28 days after irradiation. The TNF-alpha in the mice intestinal tissue was detected by means of immunohistochemical staining and image analysis. RESULTS: In normal control mice, TNF-alpha was expressed in the cytoplasm of macrophages in intestinal villus interstitium, submucosa and lymph tissue. After 2.5Gy neutron radiation, TNF alpha was decreased progressively within 2 days, increased obviously in macrophages and crypt cells during the 3rd-7th day, reached the peak at the 5th day and recovered to normal level at the 14th day. TNF-alpha was decreased progressively within 4 days after 4.0 and 5.5Gy neutron and 12Gy gamma ray irradiation. TNF-gamma was increased obviously in 6-12 hours, decreased on the first day, increased at the 2nd-5th day, peaked at the third day and recovered at the 10th day after 5.5Gy gamma ray irradiation. CONCLUSION: Neutron and gamma ray radiation induce different expression profile of endogenous TNF-gamma in small intestine, which may be related with the pathologic courses of irradiation induced damage and repair of intestine. PMID- 16256027 TI - [The relationship between TNF-alpha and the injury of renal tubular cells caused by anoxia/reoxygenation]. AB - AIM: To investigate the relationship between TNF-alpha and renal tubular cell injury caused by anoxia/reoxygenation. METHODS: Human renal proximal tubular cell line HK-2 was used as model. Anoxia/reoxygenation were produced by covering/de covering the cell culture with liquid paraffin wax. The level of TNF-alpha and the activity of lactate dehydrogenase(LDH) in the culture medium was determined by radioimmunoassay(RIA) and biochemical methods, respectively. Trypan blue exclusion was used to measure cell viability. RESULTS: Anoxia/reoxygenation could increase TNF-alpha level and LDH activity, but decrease viability of HK-2 cells. TNF-alpha level was positively correlated with LDH activity (r=0.89, P<0.05) and negatively with the cell viability (r=-0.91, P<0.05). CONCLUSION: TNF-alpha induced by anoxia/reoxygenation may participate in the process of renal tubular cell injury. PMID- 16256028 TI - [Cloning and expression of human interleukin 24 gene and study on its activity of inducing tumor cells apoptosis in vitro]. AB - AIM: To clone and express IL-24 gene in E.coli and to study its antitumor activity to induce apoptosis of tumor cells after purification and refolding. METHODS: The total RNA was extracted from peripheral blood mononuclear cells (PBMC) and induced by ConA. The human IL-24 gene was amplified by RT-PCR before subcloned into expression vector pET32a(+) and expressed in E. coli. The fusion protein was identified by Edman and purified in chelating sepharose fast flow chromatography and refolding. The activity of inducing tumor cells apoptosis was observed by MTT colorimetry and morphological study in vitro. RESULTS: The obtained human IL-24 gene was identical with that included in GenBank. The recombinant IL-24 could be expressed in E. coli by IPTG induction as inclusion body (35 kD) with purity over 95%. The apoptosis of MCF-7 cells was induced by Trx-IL-24 protein. CONCLUSION: The fusion protein can be highly expressed in E. coli and the obtained protein with high purity can induce tumor cells apoptosis. PMID- 16256030 TI - [Enhanced prokaryotic expression of a recombinant Lagurus lagurus zona pellucida subunit 3 with codon optimization]. AB - AIM: To improve prokaryotic expression level of Lagurus lagurus zona pellucida 3(LZP3) gene. METHODS: The rare codons of LZP3 gene fragment were turned into the most frequently used codons in E. coli by overlap PCR. LZP3 gene and mutated LZP3 (LZP3m) gene were cloned into expression vector pGEX4T-1 respectively, and transformed into E. coli BL21(DE3). RESULTS: The expression level of LZP3m was notably higher than that of LZP3. CONCLUSION: The expression of LZP3 gene in E. coli could be successfully improved by using codon optimization. PMID- 16256029 TI - [The effects of resveratrol on the activation, proliferation and cytokine expression of murine T lymphocytes]. AB - AIM: To study the effects of resveratrol (RSV) on the activation, proliferation and cytokine expression of murine T lymphocytes. METHODS: Murine T lymphocytes were isolated from lymph nodes and cultured in vitro. The lymphocytes were pre treated with RSV for 1 h prior to activation with PDB plus ionomycin. (3)H-TdR incorporation was used to detect T cell proliferation. IL-2 and IFN-gamma mRNA were detected by RT-PCR, IL-2 and IFN-gamma proteins were detected by intracellular cytokine staining with flow cytometry. RESULTS: RSV can inhibit the proliferation and expression of IL-2 and IFN-gamma of T lymphocytes. CONCLUSION: The immunosuppressive action of RSV may be related with its inhibitory effects on T cell activation, proliferation and cytokine expression. PMID- 16256031 TI - [Identification of HTNV-NP-specific T lymphocyte epitopes and analysis of the epitope-specific T cell response]. AB - AIM: To identify the nucleocapsid protein of Hantaan virus (HTNV-NP)-specific T lymphocyte epitopes and analyze the epitope-specific T cell response during hemorrhagic fever with renal syndrome (HFRS). METHODS: T lymphocyte epitopes and frequencies of epitope-specific T cells were determined by ELISPOT using PBMCs from HFRS patients stimulated by individual or mixture of overlapping 15-mer peptides spanning the amino acid sequence of HTNV-NP. RESULTS: Out of 10 peptide mixtures, 8 elicited strong HTNV-NP-specific responses in 18 of 47 HFRS patients, and the T cell response was found at early stage of HFRS. Moreover, 17 HTNV-NP specific T lymphocyte epitopes were identified in 11 patients, and most epitopes were clustered near the center of NP in linear structure. Among them, 14 T lymphocyte epitopes were described for the first time. CONCLUSION: HTNV-NP specific T cell response can be elicited at early stage of HFRS and T lymphocyte epitopes mainly located in the center of NP, suggesting that it may play an important role in immune protection during HTNV infection. PMID- 16256032 TI - [Expression of human T-cell immunoglobulin mucin 3(TIM-3) on eukaryotic cells and establishment of stable transfectant cell line]. AB - AIM: To construct eukaryotic expression vector of human T-cell immunoglobulin mucin 3(TIM-3) and transfect mammalian cells to establish stable cell line. METHODS: The whole coding region of TIM-3 was amplified by PCR and inserted into eukaryotic expression vector pIRES2EGFP. The recombinant plasmid was transfected into mammalian cells by Lipofectamine. The expression product was analyzed by flow cytometry and Western blot. The stable transfectant was screened and established by flow cytometry and selective medium. RESULTS: COS-7 and CHO cells were transfected with recombinant plasmid by Lipofectamine. The expression speciality was identified by Flow cytometry and Western blot. The stable transfectant of CHO cell line was established. CONCLUSION: The whole coding region of TIM-3 was successfully subcloned into eukaryotic expression vector and expressed on the surface of mammalian cells. The stable transfectant of CHO cell line was established. PMID- 16256033 TI - [Preparation of humanized anti-digoxin scFv and diabody]. AB - AIM: To prepare humanized anti-digoxin(Dig) antibodies(single chain Fv, scFv) from a human phage antibody library and construct anti-Dig diabody vector. METHODS: A human phage antibody library was panned against immobilized Dig for four times, and the specificity of the selected scFv expression clones was identified by ELISA. Positive clones against Dig were analyzed by DNA fingerprint and sequencing. A clone which had high affinity to Dig was selected to construct a diabody vector. Diabody was secreted from E. coli by IPTG induction, and the specificity was also identified by ELISA. RESULTS: After four rounds of panning, four specific humanized anti-Dig antibodies(scFv) were obtained. DNA fingerprint and sequencing analysis proved that they had different Ab-encoding genes. The V(L) of the 4 expression clones belonged to lambda subgroup 1 and V(H) belonged to 3 and 4 subgroups, respectively. A scFv clone was picked out to construct diabody vector. The prepared humanized diabody reacted with Dig specifically. CONCLUSION: Humanized anti-Dig antibodies(scFv) have been got by using phage display. A diabody has been obtained from scFv. These humanized anti-Dig antibodies may be used for the diagnosis and therapy of Dig toxication. PMID- 16256035 TI - [Construction, structure prediction and phylogenetic analysis of murine scFv gene against human cervical cancer]. AB - AIM: To construct the gene of murine single-chain Fv fragment (scFv) against human cervical cancer and to predict the physical and chemical characteristics, secondary structure, tertiary structure and the molecular evolution of the scFv using computer-assisted modeling. METHODS: The variable region gene of the heavy and light chains were amplified respectively using recombinant DNA techniques from CSA125 hybridoma cells that can secret monoclonal antibodies with high activity and specificity against human cervical cancer. These two fragments were then spliced together through a flexible linker to scFv against human cervical cancer by using splicing overlap extension (SOE) PCR. The scFv gene, named CSAs 1, was cloned into the vector pMD18-T. The physical and chemical characteristics secondary and tertiary structure and the phylogenetic tree were predicted using Internet and corresponding softwares. RESULTS: The whole scFv gene was cloned successfully and encoded 278 amino acids. Theoretically, the CSAs-1 scFv was relative molecular masses 28,800, the isoelectric point (pI) was 7.215. PHDsec indicated that CSAs-1 was alpha+beta globin. There were many protein kinase C phosphorylation sites and casein kinase II phosphorylation sites in the V(H) and V(L) domains. The computer graphic modeling indicated the linker was isolated from V(H) and V(L). The V(L), as well as the V(H), was involved in composing the hydrophobic "pocket" which was beneficial to the antigen binding. Phylogenetic tree analysis indicated that V(H) genes from various vertebrate species formed three clusters that corresponded to three V(H) groups and that V(L) sequences from each species had a level of diversity similar to that of their V(H) genes. CSAs-1 V regions belong to A group. CONCLUSION: The construction and analysis of a molecular model of scFv laid the foundation for the further research into genetic engineering antibody and the mechanism of antigen-antibody interaction. PMID- 16256034 TI - [Construction and expression of anti-keratin single-chain Fv antibody]. AB - AIM: To construct an expression vector of human anti-keratin scFv and express functional scFv in E. coli. METHODS: The V(H) and V(L) genes were amplified by PCR and cloned into a scFv expression vector, which was expressed in E. coli. The expression of scFv was tested by SDS-PAGE; The antigen-binding characteristics of scFv were tested by ELISA. RESULTS: The V(H) and V(L) genes were cloned and expressed in E. coli successfully. The expressed scFv retained the antigen binding characteristics of the parental antibody. CONCLUSION: A functional anti keratin scFv was obtained. PMID- 16256036 TI - [Prokaryotic expression and purification of antifreeze protein DAFP and preparation of its antiserum]. AB - AIM: To prepare the Dendroides canadensis antifreeze proteins (DAFP) specific mouse antiserum. METHODS: According to the published antifreeze protein gene (gi: 2737939) from Dendroides canadensis in GenBank, one isoform of the gene was chemically synthesized and cloned into the expression vector pGEX-4T-1 and pMAL p2x respectively and expressed in E.coli. The expressed fusion protein was purified through Glutathione Sepharose 4B column, and then used to immunize the mice for preparing the specific the antibody. The titer and specificity of the antibody were analyzed by ELISA and Western blot, respectively. RESULTS: SDS-PAGE analysis showed that the DAFP was expressed in E.coli. The relative molecular masses of expressed fusion protein was 38,000. The titer of antibody was 1:5,000. Western blot analysis showed the expressed protein of BL21/pGEX-4T-1-dafp and TB1/pMAL-p2x-dafp could react specifically with DAFP antibody. CONCLUSION: The DAFP was expressed in E.coli and its specific antibody was prepared successfully. PMID- 16256037 TI - [Expression of SARS coronavirus nucleocapsid protein and construction of its DNA vaccine]. AB - AIM: To express the nucleocapsid (N) protein of SARS coronavirus (SARS-CoV) in E. coli and construct its DNA vaccine. METHODS: The prokaryotic expression vector pQEN containing N gene was constructed and transformed into the E. coli. The recombinant N protein was then expressed and purified by Ni(2+)-NTA affinity resin. In addition, the N gene was cloned into the eukaryotic expression plasmid pSecTagB and the eukaryotic recombinant expression vector pSecN was obtained. The DNA vaccine pSecN was injected to immunize the BALB/c mice to produce the antiserum against N protein of SARS-CoV. Subsequently, the reactivity of the antiserum with recombinant N protein and SARS-CoV particles was assayed by ELISA. RESULTS: Recombinant N protein reacted strongly and specifically with the sera from immunized mice and SARS patients. Similarly, the sera of immunized mice could also react specifically with SARS-CoV particles. CONCLUSION: The recombinant N protein could be used as a good antigen to detect SARS. The DNA vaccine pSecN could also efficiently induce the production of IgG against N protein of SARS-CoV, which offered clues to the development of a potential DNA vaccine. PMID- 16256038 TI - [High expression of hepatoma associated antigen HAb18G in prokaryotic cells and study on its function in vitro]. AB - AIM: To construct prokaryotic expression vector of HAb18G, and express high level of this fusion protein in E. coli and to identify its function and immunogenicity. METHODS: The HAb18G full length cDNA from pBluescript/HAb18G was obtained by PCR and cloned into prokaryotic expression vector pRSET-C and then transformed into E. coli BL21(DE3) to induce its expression. Expressed products were analyzed by SDS-PAGE and laser thin layer scan. The purified HAb18G protein was identified by gelatin enzymogram and ELISA. RESULTS: Endonuclease digestion and DNA sequencing proved that HAb18G cDNA was cloned correctly into the expression vector. Result of SDS-PAGE showed that the relative molecular mass of the expressed product HAb18G fusion protein was 34,600, which was in accordance with predicted relative molecular mass value. Laser thin layer scan showed that the expressed product accounted for 33% of the total bacteria protein. Result of enzymogram was negative whereas the result of ELISA was positive. CONCLUSION: It was testified that the protein HAb18G has immunogenicity but no bioactivity. The high level prokaryotic expression of HAb18G lay the foundation for manufacturing the HAb18G protein in great quantities and proceeding to its relative research. PMID- 16256039 TI - [Preparation and characterization of monoclonal antibodies against AR protein]. AB - AIM: To prepare monoclonal antibodies (mAbs) against aldose reductase (AR) and compare with anti-aldose reductase-like protein (ARL-1) mAb. METHODS: The AR gene was gained by RT-PCR and inserted into pGEX-4T-1 (His)(6)C. Recombinant protein GST-AR was used to immunize BALB/c mouse. MAb was prepared by hybridoma technique and detected by ELISA and Western blot. Simultaneously, according to the analysis of AR by software Clustalx and Antheprot, GST-dAR(80-142 aa), GST-dA1(1-79 aa), GST-dA2(80-99 aa), GST-dA3(111-142 aa) and GST-dA4(143-316 aa) were expressed in E. coli Rosetta. All the proteins were used to analyze the binding sites of the mAb and AR protein by Western blot. RESULTS: Three clones secreting anti-AR mAb were obtained. They were all of IgG1. And the titer of mAb in ascites was 1:400,000 while in cell culture was 1:10,000. All of the three anti-AR mAbs reacted to GST-AR and proteins of placenta tissues and had no cross-reaction to GST-ARL-1 and GST protein. And the three anti-AR mAb could recognize GST-dA1, GST dA3 and GST-dA4, respectively. CONCLUSION: Three specific mAbs against AR are obtained and recognize the 1-79, 111-142, 143-316 amino acid sites of AR, respectively. The anti-AR mAb, together with the anti-ARL-1 mAb, may be a useful tool for further study of the function of AR and ARL-1 and the relationship between the two proteins and relevant diseases as well as for the epidemiological investigation. PMID- 16256040 TI - [Effect of neutralizing anti-VEGF antibody on genesis of murine type II collagen induced arthritis]. AB - AIM: To understand the effect of neutralizing anti-vascular endothelial growth factor (VEGF) antibody on the genesis of murine type II collagen (CO II)-induced arthritis (CIA). METHODS: mice DBA/1J (8-10 weeks) were given intradermal injection chick tape II collagen to set up the murine CIA model. The incidence of CIA and arthritis index were measured and the pathologic changes of articular tissue were observed after the injection of neutralizing anti-VEGF antibody. RESULTS: The anti-VEGF antibody could notably inhibit the genesis and severity of arthritis (P<0.05) at the early stage of CIA formation; while it made no difference after CIA was completely formed. CONCLUSION: Anti-VEGF neutralizing antibody can significantly inhibit synovial fibroblast proliferation, neovascularization and angeitis genesis, which made it a promising agent for RA treatment. PMID- 16256041 TI - [The in situ analysis of cytokine mRNA expression in immunological microenvironment about non-small cell lung cancer]. AB - AIM: Using Non-small cell lung cancer (NSCLC) as subject, to explore the characteristics of immune response in immunological microenvironment at the tumor site and its effect on anti-tumor immunity. METHODS: Using in situ hybridization (ISH), the expressions of IL-2, INF-gamma, IL-12 (p40), IL-18, IL-4, IL-10, TGF beta1, IL-1, IL-3, IL-8, GM-CSF, TNF-alpha and TGF-alpha mRNAs in lymphocytes and tumor cells from five fresh pleural effusion samples and 18 tumor tissue samples of NSCLC patients were detected by in situ hybridization with digoxin end-labeled oligonucleotide probe, using 5 tuberculous pleurisy patients as control. RESULTS: In pleural effusion and tumor tissue of NSCLC patients, the expression levels of IL-4, IL-10, TGF-alpha, and TGF-beta1 mRNAs were significantly higher than those of IL-2, IL-12, IL-18 and INF-gamma mRNAs. In contrast, the analysis of tuberculous pleural effusion samples revealed lower levels of above-mentioned cytokine mRNA. There was no significant difference among expression levels of these cytokine mRNAs. CONCLUSION: The expressions of type II cytokine mRNAs and immunosuppressive cytokine mRNAs in pleural effusion and tumor tissue of NSCLC patients occupied a dominant position, suggesting that the immunological microenvironment about tumor be in an immunosuppressive state. The present study has contributed to a better understanding the mechanisms of tumor escape and provides important experimental basis for working out the scheme for an effective immunomodulatory treatment of NSCLC patients. PMID- 16256042 TI - [Establishment of the transfected cell line expressing the mouse OX40 gene and its potency to induce differentiation of B cells]. AB - AIM: To establish HUVECs line expressing mouse OX40(CD134) and to study its promotive effect on proliferation of B cells. METHODS: The cDNA fragment encoding mouse OX40 was obtained from the total RNA of ConA-activated lymphocytes of thymus by using RT-PCR and cloned into pUCm-T vector. The cDNA was then inserted into the eukaryotic expression vector pIRES2-EGFP. The recombinant vector was transfected into HUVECs with lipofectin reagent, and the positive cellular clones were selected by G418. Expression of mouse OX40 in the transfected cells was analyzed by FCM. The differentiation of B cells in vitro induced by OX40 signal was studied by means of (3)H-TdR method. RESULTS: The cDNA fragment in the recombinant plasmid was consistent with the reported mouse OX40 cDNA in GenBank, which was confirmed by DNA sequencing, PCR and enzyme digestion. The HUVECs stably expressing the mouse OX40 were successfully cloned. The transfected cells promoted the differentiation of B cells in vitro and there existed a synergic effect between OX40/OX40L and CD40/CD40L signals. CONCLUSION: Transfected cell line expressing the mouse OX40 gene is established successfully. OX40 enhances the proliferation of B cells. PMID- 16256043 TI - [The effect of hIL-4 on the expression of E-selectin and ICAM-1 on bovine aortic endothelial cells]. AB - AIM: To investigate the effect of human interleukin-4 (hIL-4) on the expression of E-selectin and ICAM-1 on bovine aortic endothelial cells(BAEC) activated with TNF-alpha. METHODS: BAEC were cultured in different concentrations of hIL-4 for 2 h followed by treatment with 4 microg/L TNF-alpha for 6 h or 18 h. The expression of E-selectin and ICAM-1 on BAEC was detected by cell-ELISA. The effect of hIL-4 on viability of BAEC was detected by MTT colorimetry. RESULTS: Pretreatment of BAEC with hIL-4 inhibited the expression of E-selectin and ICAM-1 on BAEC activated by TNF-alpha in a dose-dependent manner (P<0.01). Both TNF-alpha and hIL-4 had no effect on the viability of BAEC. CONCLUSION: hIL-4 can suppress the expression of E-selectin and ICAM-1 on BAEC activated by TNF-alpha, which may contribute to the xenotransplant immune tolerance associated with hIL-4. PMID- 16256044 TI - [Influence of IL-2 on proliferation of cultured human pituitary adenoma cells and its mechanism]. AB - AIM: To investigate the effects of IL-2 on the proliferation, cell cycle, protein kinase C(PKC) and cAMP/cGMP of cultured human pituitary adenoma cells. METHODS: MTT colorimetry, (3)H-TdR and flow cytometry were used to detect the proliferation and radioimmunoassay was used to observe the effect of IL-2 on PKC activity and cAMP/cGMP levels of cultured human pituitary adenoma cells. RESULTS: (1) IL-2 (1 x 10(4), 1 x 10(5) and 5 x 10(5) U/L) stimulated the proliferation and DNA synthesis of cultured human pituitary adenoma cells in a dose-dependent manner. (2) IL-2 (1 x 10(4), 1 x 10(5) and 5 x 10(5) U/L) decreased the ratio of pituitary adenoma cells in G(1) phase and increased the ratio in S and G(2) phase markedly. (3) Compared with control, PMA, a PKC activator, increased the activity of membrane and total PKC in human pituitary adenoma cells. However, after treatment with IL-2 (1 x 10(5) U/L), a significant increase of the activity of cytoplasmic, membrane and total PKC in human pituitary adenoma cells was observed. (4) IL-2 (5 x 10(4), 1 x 10(5) U/L) decreased the amount of cAMP in the cytoplasm of human pituitary adenoma cells, but had no effect on that of cGMP. CONCLUSION: IL-2 can stimulate the proliferation of pituitary adenoma cells through PKC and cAMP/cGMP signaling pathways. PMID- 16256045 TI - [Effects of Angelica sinensis polysaccharide on cell-mediated immunity]. AB - AIM: To investigate the effects of Angelica sinensis polysaccharide (AP) on cell mediated immunity. METHODS: The lymphocyte proliferation was tested by MTT colormetry; T lymphocyte fraction were separated with immune magnetic beads and the change of CD4(+) T lymphocyte percentage was detected by flow cytometry. RESULTS: In the range of 30-300 mg/L, the total AP and its three fractions, ie AP 0, AP-1, AP-2, AP-3, directly stimulated the proliferation of murine splenocytes, and AP-3 significantly enhanced the proliferation of lymphocytes in MLR and T lymphocytes. The percentage of CD4(+) T lymphocytes in total splenocytes was also enhanced by AP-3. CONCLUSION: AP can promote the cell-mediated immunity with T lymphocytes as one of its target cells. PMID- 16256047 TI - Risk and spectrum of diseases in travelers to popular tourist destinations. AB - BACKGROUND: Traveling to tropical regions is related to increased health risks. Travelers' diarrhea is the most frequent health problem, but the range of travel related diseases also includes potential life-threatening diseases such as malaria. The actual risk of European travelers acquiring specific infectious diseases and other hazards in the tropics is to a large extent unknown and is therefore often adopted from that of the indigenous population. The objective of this study was to elucidate the risk for travel-related diseases, symptoms, and accidents in a population of Europeans who travel to popular tourist destinations. METHODS: From July 2003 to June 2004, 794 travelers consulting the travel clinic of the Berlin Institute of Tropical Medicine were recruited for a questionnaire-based observational study before traveling to Kenya, Tanzania, Senegal, the Gambia, India, Nepal, Thailand, or Brazil. RESULTS: Overall, illness was reported by 42.9% of travelers, with 10.2% reporting more than one adverse health event. Most frequently gastrointestinal symptoms were noted (34.6%), followed by respiratory symptoms (13.7%). More than 5% experienced an accident. Travel to the Indian subcontinent nearly doubled the risk of becoming ill; travel to Thailand significantly decreased the risk. Additional risk factors were a long duration of staying abroad, young age, and traveling under basic conditions. Of all travelers, 80% did not follow the traditionally recommended dietary restrictions. Among travelers visiting malaria-endemic areas, 20% did not carry any antimalarial drugs with them, not continuous chemoprophylaxis or standby medication. CONCLUSIONS: Because of the rising travel activity, especially to tropical countries, the importance of qualified pretravel advice consultation is increasing. To improve the travelers' health, attention needs to be paid to individual risk factors, the prevention and therapy of travelers' diarrhea, malaria prophylaxis, management of respiratory illness, and personal safety. PMID- 16256046 TI - Correlation between adherence to precautions issued by the WHO and diarrhea among long-term travelers to India. AB - BACKGROUND: Travelers' diarrhea is the most common infectious disease afflicting travelers to developing countries. Most studies investigating the benefits of recommendations regarding the consumption of food and water have focused on short term travelers. We investigated the benefits of adherence to the precautions from the World Health Organization (WHO) among long-term travelers. METHODS: We asked 140 incidental travelers in India traveling for at least 2 months to complete a questionnaire about their adherence to the WHO precautions and the occurrence of diarrhea. Adherence was graded on a scale of 1 to 6 (least to most). RESULTS: The mean age of the 114 travelers whose questionnaires were eligible was 26.6 +/- 5.7 years, and the median duration of their trip was 5 months. None of them adhered strictly to the entire set of rules. The mean individual adherence was 3.4 (range 1.2-5.8). The vast majority of the travelers (83%) suffered from diarrhea. Most travelers (60%) had diarrhea for up to 3% of their journey time. Diarrhea was accompanied by fever among 18% and necessitated hospitalization in 3%. Forty-five percent indicated that they had lost traveling days due to diarrhea, for an average of 0.7% of the traveling time. We found no correlation between the percentage of traveling time with diarrhea and the following variables: adherence to the WHO recommendations, receipt of advice regarding prevention, duration of the trip, age, sex, and nationality of the travelers. CONCLUSIONS: Dietary self restraint of travelers as proposed by the WHO is both difficult to comply with and lacks a proven value for the long-term traveler to a developing country. PMID- 16256048 TI - Telemedical advice to long-distance passenger ferries. AB - BACKGROUND: Radio medical (RM) advice for seafarers and traveling passengers is important and can be crucial for the optimal medical treatment on board ships. The aim of this study was to analyze the data from consultations with passenger ferries to identify areas for possible improvements. METHODS: Data from the journals for 1 year from Radio Medical Denmark consultations with the medical officers on passenger ferries were analyzed retrospectively. RESULTS: Two hundred fourteen RM records, 73% pertaining to passengers and 27% for crew members, were analyzed. Passenger patients were generally older and more seriously ill than patients among the crew. A high number of potential and life-threatening medical conditions such as angina pectoris was seen among the passengers, and nine of these patients were evacuated by helicopter. Sixty-three percent (n = 135) of the calls related to pain complaints, and more than half of these involved severe or considerable pain. Only acetaminophen (paracetamol) and opioids were in the ferry medicine chest. At least 77 patients would have benefited from use of nonsteroidal anti-inflammatory drugs. CONCLUSIONS: The paramedical assistance and the medicine chest contents were considered insufficient in several cases. Passengers and crew members with chronic illnesses should be thoroughly prepared and advised before their travels. PMID- 16256049 TI - Evaluation of plasma lactate as a parameter for disease severity on admission in travelers with Plasmodium falciparum malaria. AB - BACKGROUND: Rapid immunochromatographic dipstick assays are used increasingly in many tropical and Western countries as a tool to diagnose Plasmodium falciparum malaria. However, these tests do not provide any information about the severity of the infection. We evaluated the usefulness of plasma lactate as a parameter for disease severity on admission in imported P. falciparum malaria. METHODS: In a cohort of 61 nonimmune travelers with imported P. falciparum malaria, plasma lactate levels on admission were related to the severity of the infection. Results from 12 of 61 patients fulfilled the criteria of severe malaria. RESULTS: Logistic regression analysis showed that a plasma lactate level above the upper range of normal was associated with an odds ratio of 31 (95% CI 6-164) for severe malaria. As a continuous variable, a 1 mmol/L increase in plasma lactate level was associated with an odds ratio of 12 (95% CI 3-50) for severe malaria. The sensitivity of an increased plasma lactate level on admission for severe malaria was 67% with a specificity of 94%. CONCLUSIONS: A timely determination of plasma lactate on admission may be helpful in the assessment of disease severity in travelers with imported P. falciparum malaria. An increased plasma lactate level should raise suspicion of a severe P. falciparum malaria infection, in particular when concomitant infections are not considered likely. PMID- 16256050 TI - Schistosomiasis and US Peace Corps volunteers in Tanzania. AB - BACKGROUND: Schistosoma haematobium and Schistosoma mansoni are endemic to East Africa. US Peace Corps volunteers (PCVs) serving in Tanzania were not experiencing symptoms of infection, but in 1998 when comprehensive testing was instituted, more than one-quarter were found to be infected with schistosomiasis at the close of their 27-month tour of service. METHODS: An education campaign was instituted by the in-country Peace Corps Medical Officers. At their close of service, all PCVs had a medical examination and blood, urine, and stool samples were tested for schistosomiasis. PCVs also answered a survey that included questions about their knowledge of the blood fluke and the disease, symptoms, where and how many times they had gone swimming, and what precautionary measures they had taken. RESULTS: PCVs in Tanzania are most susceptible to infection from recreational activities within the Lake Victoria ecosystem. After an education campaign, incident cases dropped quickly. Vigorous toweling after exposure may have decreased the risk of disease for those PCVs who went swimming in freshwater. Although schistosomiasis is common in Tanzania and PCVs are highly susceptible, it is possible for them to spend their entire tour without contracting the disease, even if they live by Lake Victoria. CONCLUSIONS: People should be encouraged not to swim in Lake Victoria. The possibility that vigorous toweling immediately after exposure to infected waters is a useful protective measure warrants further investigation. PMID- 16256051 TI - Epidemiology of typhoid fever in Mauritius. AB - BACKGROUND: The epidemiology of typhoid fever in Mauritius was studied to determine whether there was any need for tourists visiting Mauritius to be vaccinated against the disease, and where Mauritians with typhoid fever had been infected. Data on antibiotic susceptibility of Salmonella typhi isolates from Mauritius were also analyzed. METHODS: Since 1997 every time S. typhi is isolated from blood cultures at our laboratory, an epidemiologic inquiry is conducted to determine the likely origin of the infection and the outcome of treatment, and the information collected is recorded. Results of antibiotic susceptibility testing are also noted. Data recorded on cases between 1997 and 2004 were reviewed and analyzed. RESULTS: S. typhi was isolated on 25 occasions during the 8-year period. The infection was likely to have been acquired in Mauritius in only 6 cases (24%). Another 6 cases (24%) occurred in expatriate workers from the Indian subcontinent. Of the 13 Mauritians (52%) who probably acquired the infection abroad, 11 had a history of recent travel to India. Thirteen of 14 S. typhi isolates from cases acquired in India were resistant to nalidixic acid. Of the 6 cases acquired in Mauritius, 4 occurred in children under 12 years and 1 was caused by a multiply resistant strain. Twenty-two patients made an uneventful recovery. One death was indirectly caused by typhoid fever, and there was 1 case each of intestinal perforation and relapse. CONCLUSIONS: In Mauritius typhoid fever is mainly an imported disease, but indigenous cases of the illness occur rarely and sporadically. Travelers to Mauritius need not be vaccinated against typhoid fever as the risk of acquiring the disease in the country is negligible. Mauritians traveling to India must be made aware of the risk of typhoid fever and of preventive measures. Ceftriaxone should be used as the initial first-line treatment of infection acquired in India. PMID- 16256052 TI - Enteric fever in Israeli travelers: a nationwide study. AB - BACKGROUND: Enteric fever (EF) has become a travel-related disease in industrialized countries. The possible effects of vaccination on typhoid epidemiology in travelers are unknown. We compared the incidence and clinical and microbiologic findings in travelers returning with EF, according to pretravel vaccination status and vaccine type. METHODS: We performed a nationwide descriptive analysis of EF incidence in Israeli travelers; EF is a notified disease in Israel. Data from 1995 through 2003 were evaluated; all cases of EF acquired during recent travel (6 wk) were included. From 1995 to 1999, the Ty21a oral vaccine was used exclusively in Israel. It was replaced with the Vi vaccine from 2000 to 2003. Patients with pretravel typhoid vaccination were compared with unvaccinated patients, and according to vaccine type. RESULTS: Seventy-eight cases met our criteria. The causative agents were Salmonella typhi in 79.5% and Salmonella paratyphi A in 20.5%; 74.4% were acquired by travelers to the Indian Subcontinent. S. paratyphi A accounted for 10.5% of cases among Ty21a vaccinees as compared with 47.4% among Vi vaccinees (p = .02). For the Indian Subcontinent, the general attack rate of S. typhi and S. paratyphi A during the period of vaccination with Ty21a was 2.37 in 10,000 and 0.26 in 10,000 travelers, respectively, whereas during the period of vaccination with Vi, the attack rate was 1.40 in 10,000 and 0.79 in 10,000. There were no deaths; however, more complications and relapses occurred in the S. paratyphi A group. CONCLUSIONS: Among Israeli travelers S. typhi infection is declining whereas S. paratyphi A is increasing, with most cases occurring in vaccinated travelers. Prior typhoid vaccination did not modify the course of the disease. S. paratyphi A infection in travelers is not milder than S. typhi infection. Although this is not a prospective, controlled, randomized trial, it appears that the Ty21a vaccine may be less effective for S. typhi but may offer some protection against S. paratyphi A. Sequential vaccination with the available oral and Vi vaccine may merit consideration. A more effective vaccine for S. typhi and S. paratyphi A is urgently needed. PMID- 16256053 TI - Imported Brucellosis associated with Plasmodium falciparum malaria in a traveler returning from the tropics. AB - We report a microbiologically confirmed case of Brucella melitensis and Plasmodium falciparum malaria coinfection in a febrile traveler returning from Chad, Africa. The patient had been doing veterinary research in rural Chad; during that time she took no antimalarial chemoprophylaxis. Our report highlights the importance of blood cultures as well as malaria smears in febrile travelers returning from the tropics. PMID- 16256054 TI - Myiasis with Dermatobia hominis in a traveler returning from Costa Rica: review of 33 cases imported from South America to Japan. AB - We experienced a case of Dermatobia hominis infestation from South America. Here we discuss imported cases of D. hominis in Japan with literature. PMID- 16256055 TI - Acute hepatitis and atovaquone/proguanil. AB - A 31-year-old healthy man developed acute hepatitis after receiving atovaquone (250 mg) and proguanil (100 mg) for malaria prophylaxis daily for 25 days. Although atovaquone/proguanil is generally well-tolerated, this case highlights the hepatotoxic potential with considerable morbidity and should alert physicians to this harmful side effect. PMID- 16256056 TI - Penicillium marneffei infection in an immunocompromised traveler: a case report and literature review. AB - Penicillium marneffei has emerged as an important opportunistic pathogen in Southeast Asia during the human immunodeficiency virus (HIV) epidemic. We report a case of disseminated P. marneffei in a person with previously undiagnosed acquired immunodeficiency syndrome (AIDS) who traveled to Southeast Asia, illustrating the importance of considering this diagnosis in immunocompromised travelers. PMID- 16256058 TI - Schistosoma intercalatum: a missed diagnosis? PMID- 16256059 TI - Travel- associated infectious diseases. Selected bibliography. PMID- 16256057 TI - Acute meningitis owing to phlebotomus fever Toscana virus imported to France. AB - Acute lymphocytic meningitis, seen in France in summer, is often due to enteroviruses. Arboviruses as West Nile and tick-borne encephalitis do exist in Europe, but other viruses are rarely considered in patients unless they have had recent tropical travel. Toscana virus infection, which is endemic, especially in Italy,1-3 has been documented in some European travelers returning from Italy,4-6 but surprisingly was not documented in France until recently.7 We report a case of meningitis caused by Toscana virus imported to France. PMID- 16256061 TI - The persistence, dissemination, and visceralization tendency of Leishmania major in Syrian hamsters. AB - I monitored the persistence, dissemination, and the possible visceralization tendency of Leishmania major, the causative parasite of cutaneous leishmaniasis in North Africa and the Middle East in Syrian hamsters (Mesocricetus auratus). Hamsters were inoculated subcutaneously in the hind footpad, with 1 x 10(6)L. major metacyclic promastigotes and were sacrificed at months 1, 3, 6 and 10 post infection (pi). Skin lesions, blood, spleens, livers and kidneys were screened by both Giemsa-stained smears and a polymerase chain reaction (PCR) for detection of L. major amastigotes. A few weeks pi, 61.7% of the inoculated hamsters developed a cutaneous lesion only at the inoculation site, while 38.3% of them developed non-self-healed lesions at sites distant from the inoculation site. PCR identified all the positive stained smears as well as false-negative ones, indicating that PCR was more sensitive than stained smears. The results confirmed the dissemination and persistence of L. major amastigotes in all tissues examined, except the kidneys, for a period extending to 10 months, only in those hamsters suffering from disseminated cutaneous lesions. Parasite DNA was detected in the bloods starting from the first month pi and starting from month 3 pi in the spleens and livers. Some, but not all, the animals with disseminated infections proved to be positive for parasite DNA in their organs. Persistence of the L. major amastigotes in the tissues differed from those of other species causing visceral diseases. These findings demonstrate a possible visceralization tendency for L. major previously recorded for L. tropica and L. mexicana. PMID- 16256062 TI - In vitro antimalarial activity of flavonoid derivatives dehydrosilybin and 8 (1;1)-DMA-kaempferide. AB - Multidrug-resistant Plasmodium falciparum strains are an increasing problem in endemic areas and are partly responsible for the worsening malaria situation around the world. New cheap and effective compounds active in combination with available drug in the field are urgently needed. The aim of this work was to explore the potential antiplasmodial effect of flavonoid derivatives on parasites growth in vitro. In vitro antiplasmodial activity of dehydrosilybin and 8-(1;1) DMA-kaempferide has been evaluated by real time PCR for five P. falciparum strains. Both revealed significative antimalarial activity against the different strains. Since this drug family has been largely used and well-tolerated in humans, flavonoid derivatives could be in the near future associated with already available drugs in order to delay the spread of P. falciparum resistance. PMID- 16256063 TI - An unexpected diterpene cyclase from rice: functional identification of a stemodene synthase. AB - We have cloned a novel diterpene synthase (OsKSL11) from rice that produces stemod-13(17)-ene from syn-copalyl diphosphate. Notably, this gene sequence was not predicted from the extensive sequence information available for rice, nor, despite extensive phytochemical investigations, has this diterpene or any derived natural product previously been reported in rice plants. OsKSL11 represents the first identified stemodene synthase, which catalyzes the committed step in biosynthesis of the stemodane family of diterpenoid natural products, some of which possess antiviral activity. In addition, OsKSL11 is highly homologous to the mechanistically similar stemarene synthase recently identified from rice, making this pair of diterpene cyclases an excellent model system for investigating the enzymatic determinants for differential product outcome. The unexpected nature of this cyclase and its product parallels recent observations of previously unrecognized natural products metabolism in Arabidopsis thaliana, suggesting that many, if not all, plant species will prove to have extensive biosynthetic capacity. PMID- 16256064 TI - Heterologous expression and site-directed mutagenesis of an ascorbate-reducible cytochrome b561. AB - Cytochromes b561 (Cyts b561) are ubiquitous membrane proteins catalyzing ascorbate-mediated trans-membrane electron transfer. A heterologous expression system in Saccharomyces cerevisiae was developed to study their structure function relationship. Recombinant mouse chromaffin granule Cyt b561 (CGCytb) shows spectral characteristics, ascorbate reducibility, and redox potentials identical to that of the native bovine protein. Moreover, the reconstituted recombinant protein mediated trans-membrane electron transport with kinetic characteristics similar to that of bovine CGCytb. Site-directed mutant analysis supports the presence of two hemes coordinated by the highly conserved His pairs H52/H120 and H86/H159. Reduction of CGCytb by ascorbate showed biphasic kinetics (Kd1: 0.016 +/- 0.005 mM, Kd2: 1.24 +/- 0.19 mM). Mutation of a well-conserved Arg residue (R72) abolished high affinity CGCytb reduction by ascorbate, indicating that this residue may be critical for substrate binding. On the other hand, mutation of a Lys previously suggested to play a role in ascorbate binding (K83), did not affect the ascorbate-mediated reduction of the protein. PMID- 16256065 TI - AtLACS7 interacts with the TPR domains of the PTS1 receptor PEX5. AB - Long-chain acyl-CoA synthetases (LACSs) activate fatty acids for further metabolism and are encoded by a multi-gene family in Arabidopsis. AtLACS6 possesses a type 2 (PTS2) peroxisomal targeting sequence, whilst AtLACS7 has both a type 1 and type 2 peroxisomal targeting sequence. AtLACS7 was used as bait in a yeast two-hybrid screen. Multiple clones of the PTS1 receptor PEX5 were isolated. Quantitative beta-galactosidase assay indicated that full-length PEX5 interacts with AtLACS7 with higher affinity than the TPR domains alone. The interaction between PEX5 and AtLACS7 was confirmed by co-immunoprecipitation and shown to be specific for the PTS1, therefore the AtLACS7 PTS1 is accessible to bind PEX5 in the full-length AtLACS7 protein. The expression profile of AtLACS6, AtLACS7, AtPEX5, and AtPEX7 revealed that AtLACS6 and 7 have distinct patterns of expression and we speculate that the possession of two targeting signals may be advantageous for the import of AtLACS7 when receptors may be limiting. PMID- 16256066 TI - Influence of the growth at high osmolality on the lipid composition, water permeability and osmotic response of Lactobacillus bulgaricus. AB - Changes in water permeability and membrane packing were measured in cells of Lactobacillus bulgaricus and in vesicles prepared with lipids extracted from them. The osmotic response of whole cells and vesicles is compared with the one of bacteria grown in a high osmolal medium. Both bacteria and vesicles, behave as osmometers. This means that the volume decrease is promoted by the outflow of water, driven by the NaCl concentration difference, arguing that neither Na+ nor Cl- permeates the cell or the lipid membrane in these conditions. Therefore, the volume changes can be correlated with the rate of water permeation across the cell or the vesicle membranes. The permeation of water was analyzed as a function of the lipid species by measuring the volume changes and the saturation ratio of the lipids. To put into relevance the membrane processes, the permeation properties of lipid vesicles prepared with lipids extracted from bacteria grown in normal and high osmolality conditions were also analyzed. The permeation response was correlated with the physical properties of the membrane of whole cells and vesicles, by means of fluorescence anisotropy of diphenyl hexatriene (DPH). The modifications in membrane properties are related with the changes in the membrane composition triggered by the growth in a high osmolal medium. The changes appear related to an increase in the sugar content of the whole pool of lipids and in the saturated fatty acid residues. PMID- 16256067 TI - Protein phosphatase-1 inhibitor-3 is co-localized to the nucleoli and centrosomes with PP1gamma1 and PP1alpha, respectively. AB - In this study, we show that protein phosphatase-1 (PP1) inhibitor-3 (Inh3) is localized to the nucleoli and centrosomes in interphase HEK 293 cells. Inh3 exhibited a specific co-localization to the nucleoli with PP1gamma1, and to the centrosomes with PP1alpha. These findings indicate that Inh3 may act as a modulator of PP1 functions in the processes of cytokinesis, as well as of nucleolar events. The specificity of the interaction of Inh3 with the PP1 isoforms was also demonstrated in vitro, where Inh3 co-immunoprecipitated with PP1alpha and PP1gamma1, but not with PP1beta. The nuclear localization signal of Inh3 was identified as a N-terminal basic cluster (33RKRK36), while nucleolar localization was shown to be dependent on a C-terminal basic cluster (94HRKGRRR100). The importance of the individual basic residues was quantitatively assessed by site-directed mutagenesis and a novel use of laser scanning cytometry. PMID- 16256068 TI - Role of lysine residues in membrane anchoring of saposin C. AB - Molecular dynamics (MD) simulations of the N-terminal region of saposin C, containing amino acid residues 4-20 (saposin C4-20), were performed over 2.5 ns in 1,2-dioleoyl-sn-glycero-3-phosphoserine (DOPS) and 1,2-dimyristoyl-sn-glycero 3-phosphocholine (DMPC) monolayers. The simulations revealed several strong specific interactions of lysine 13 (Lys13) and lysine 17 (Lys17) in saposin C4-20 with the anionic phospholipids, which are required for membrane anchoring of the peptide. Membrane anchoring of saposin C4-20 facilitates saposin C-induced liposomal membrane fusion. Substitutions of Lys13 or Lys17 with alanine or glutamic acid led to a substantial loss of saposin C's fusogenicity. However, arginine replacement of Lys13 or Lys17 caused a partial loss of saposin C's fusogenic activity. The membrane anchoring of saposin C was altered in the presence of 0.4 M sodium chloride. Differential salt effects on Lys-mutant saposin Cs were observed using Trp fluorescence analysis. Low salt concentration had a more significant impact on Lys-mutant saposin C with a negatively charged amino acid residue replacement than those mutants with a positively charged or neutral residue replacement. These results indicate that positively charged amino acids at positions 13 and 17 are required for the fusogenic function of saposin C. In addition, the side-chain structure of lysine is crucial to the precise membrane anchoring which is necessary for the total fusion activity of saposin C. The MD simulations and vesicle size measurements of lysine-mutant saposins confirm the importance of the two lysine residues in saposin C4-20 for saposin C induced fusion of negatively charged phospholipid membranes. PMID- 16256069 TI - Altered expression of neuronal nitric oxide synthase in the duodenum longitudinal muscle-myenteric plexus of obesity induced diabetes mouse: implications on enteric neurodegeneration. AB - Type 2 diabetes caused by obesity shows autonomic neuropathy. Molecular mechanism involved in enteric neurodegeneration is not clear. Neuronal nitric oxide synthase (nNOS) is one of the important agents involved in gastrointestinal function. Therefore, expression of nNOS in the duodenum LM-MP of type 2 diabetes model mouse was studied. Real time RT-PCR analysis showed reduction in nNOS expression in male diabetic LM-MP compared to male control. In contrast, female diabetic LM-MP had high level of nNOS mRNA compared to female control. Western blot determination of LM-MP showed reduction in nNOS protein in male diabetic LM MP and high level of nNOS in female diabetic LM-MP compared to their respective controls. Expression of nNOS observed by Western blot was further confirmed by nNOS immunostaining of the mouse duodenum. TUNEL staining of mouse duodenum showed apoptosis in male diabetic enteric neurons. These studies suggest that nNOS expression in LM-MP varies with gender during early stage of type 2 diabetes. In addition, reduced expression of nNOS is likely to contribute to apoptosis seen in the enteric neurons of male type 2 diabetic mice. PMID- 16256070 TI - Malignant transformation of an epithelial cell by v-Src via tv-a-mediated retroviral infection: a new cell model for studying carcinogenesis. AB - Most human cancers are of epithelial origin, but many cell culture models for the study of cancer-causing genes use fibroblasts. In addition, efficient delivery and stable expression of foreign genes into non-transformed cell lines are often difficult. To address both questions, we here established a non-transformed rat kidney epithelial RK3E cell line that constitutively expresses tv-a (receptor for subgroup A avian leukosis virus, ALV) for delivery of foreign genes via avian retroviral infection. This cell line (RK3E/tv-a) allows efficient and stable expression of either single or multiple foreign genes. Furthermore, tv-a-mediated delivery of various oncogenes (v-src, H-ras, myc or akt) leads to malignant transformation. v-src-transformed cells exhibited classical cancerous phenotypes in vitro, and induced tumor formation and lung metastasis upon injecting into immunodeficient mice. Expression profiles of downstream molecular effectors (E cadherin, beta-catenin, cyclin D1, Myc, VEGF, MMP-2, and MMP-9) in these cells correlate with characteristics of cancerous phenotypes. This new cell model serves as a useful tool to study cancer-causing genes in epithelial cell type. PMID- 16256071 TI - CIN85 regulates the ability of MEKK4 to activate the p38 MAP kinase pathway. AB - CIN85 is a multi-adaptor protein involved in different cellular functions including the down-regulation of activated receptor tyrosine kinases and survival of neuronal cells. CIN85 contains three SH3 domains that specifically bind a unique proline-arginine motif (PxxxPR) found in several CIN85 effectors. In this report, we show that the MAP kinase kinase kinase MEKK4 is a new CIN85 interacting partner. This interaction is mediated by the engagement of the SH3 domains of CIN85 to three PxxxPR motifs located within MEKK4 sequence. By disrupting this interaction we demonstrated that CIN85 binding to MEKK4 enhances the activation of MKK6 and of the downstream p38 MAP kinase following oxidative stress and growth factor stimulation. CIN85 was also shown to regulate the activation of MEKK4 by GADD45 proteins and promote multi-ubiquitination of MEKK4. Taken together, these results indicate a novel role for CIN85 in the regulation of cellular stress response via the MAPK pathways. PMID- 16256072 TI - Non-linear quantitative structure-activity relationship for adenine derivatives as competitive inhibitors of adenosine deaminase. AB - Logistic regression and artificial neural networks have been developed as two non linear models to establish quantitative structure-activity relationships between structural descriptors and biochemical activity of adenosine based competitive inhibitors, toward adenosine deaminase. The training set included 24 compounds with known k(i) values. The models were trained to solve two-class problems. Unlike the previous work in which multiple linear regression was used, the highest of positive charge on the molecules was recognized to be in close relation with their inhibition activity, while the electric charge on atom N1 of adenosine was found to be a poor descriptor. Consequently, the previously developed equation was improved and the newly formed one could predict the class of 91.66% of compounds correctly. Also optimized 2-3-1 and 3-4-1 neural networks could increase this rate to 95.83%. PMID- 16256073 TI - Biophotonic hydrogen peroxide production by antibodies, T cells, and T-cell membranes. AB - Rapidly accumulating evidence indicates that inflammatory T cells sensitively respond to their redox environment by activating signal transduction pathways. The hypothesis that T-cell receptors have the potential to catalytically transform singlet oxygen into H(2)O(2) attracted our attention since the biophysical regulation of this process would provide a new tool for therapeutically directing T cells down a preferred signaling pathway. Light dependent production of H(2)O(2) was first described in antibodies, and we reproduced these findings. Using a real-time H(2)O(2) sensor we extended them by showing that the reaction proceeds in a biphasic way with a short-lived phase that is fast compared to the slow second phase of the reaction. We then showed that Jurkat T cells biophotonically produce about 30nM H(2)O(2)/min/mg protein when pretreated with NaN(3). This activity was concentrated 4 to 5 times in T cell membrane preparations. The implications of these observations for the development of new therapeutic tools for inflammatory diseases are discussed. PMID- 16256074 TI - Caenorhabditis elegans calnexin is N-glycosylated and required for stress response. AB - Calnexin, a type I integral Ca(2+)-binding protein in the endoplasmic reticulum (ER) membrane, has been implicated in various biological functions including chaperone activity, calcium homeostasis, phagocytosis, and ER stress-induced apoptosis. Caenorhabditis elegans CNX-1 is expressed in the H-shaped excretory cell, intestine, dorsal and ventral nerve cord, spermatheca, and head and tail neurons throughout development. A cnx-1 null mutant displays temperature sensitive developmental and reproductive defects, and retarded growth under stress. Moreover, a double knockout mutant of calnexin and calreticulin exhibits additive severe defects. Interestingly, both cnx-1 transcript and protein levels are elevated under stress conditions suggesting that CNX-1 may be important for stress-induced chaperoning functions in C. elegans. Glycosidase treatment and site-directed mutagenesis confirmed that CeCNX-1 is N-glycosylated at two asparagine residues of Asn(203) and Asn(571). When transgenic animals from cnx-1 mutant were generated, a glycosylation defective construct failed to rescue phenotypes of cnx-1 mutant suggesting that glycosylation is important for calnexin's functions in C. elegans. PMID- 16256075 TI - Facial expressions of emotion reveal neuroendocrine and cardiovascular stress responses. AB - BACKGROUND: The classic conception of stress involves undifferentiated negative affect and corresponding biological reactivity. The present study hypothesized a new conception that disaggregates stress into emotion-specific, contrasting patterns of biological response. METHODS: Ninety-two healthy adults engaged in stress-challenge tasks, during which cardiovascular responses, hypothalamic pituitary-adrenocortical (HPA) axis responses (i.e., cortisol), emotional expressions (i.e., facial muscle movements), and subjective emotional experience (self-reported) were assessed. RESULTS: Pronounced individual differences emerged in specific emotional responses to the stressors. Analyses of facial expressions revealed that the more fear individuals displayed in response to the stressors, the higher their cardiovascular and cortisol responses to stress. By contrast, the more anger and disgust (indignation) individuals displayed in response to the same stressors, the lower their cortisol levels and cardiovascular responses. Individual differences in optimistic appraisals appeared to mediate these correlated patterns. CONCLUSIONS: Facial expressions of emotion signal biological responses to stress. Fear expressions signal elevated cortisol and cardiovascular reactivity; anger and disgust signal attenuated cortisol and cardiovascular reactivity, patterns that implicate individual differences in stress appraisals. Rather than conceptualizing stress as generalized negative affect, studies can be informed by this emotion-specific approach to stress responses. PMID- 16256076 TI - Use of [3H]fluorowillardiine to study properties of AMPA receptor allosteric modulators. AB - Compounds which modulate AMPA receptor function through allosteric mechanisms were examined for their effect on the binding of the agonist [3H]fluorowillardiine (FW). Benzamide-type positive modulators (ampakinestrade mark) under all experimental circumstances increased [3H]FW binding to native receptors in rat brain membranes. Benzothiadiazide drugs had more variable effects ranging from large reductions with cyclothiazide and JM-13 to increases produced by more recent compounds like PEPA, D1 and LY392098. These effects on binding were moderately influenced by the assay conditions, including temperature and the presence or absence of thiocyanate. Significant changes in agonist binding were also produced by other modulatory agents such as noncompetitive blockers (GYKI 53655, SYM 2206), polycationic compounds (spermine, Naspm, philanthotoxin) and polyanionic compounds (Evans Blue, suramin, PPNDS). EC50 values usually were similar to those from physiological studies, which validates using binding tests to assess drug potencies. Moreover, direction and magnitude of the binding change (Emax) provide information about which kinetic aspects are affected by a drug. For example, the magnitude of the binding increase produced by positive modulators was strongly correlated with their ability to slow response deactivation in excised patch recordings. Binding also provides a reliable method to examine whether interactions between agents are competitive. Thus, thiocyanate did not significantly influence the EC50 of cyclothiazide, suggesting distinct sites of action. Taken together, [3H]FW binding can yield important information about drug-receptor and drug-drug interactions for a wide range of modulatory agents. One potential limitation of [3H]FW is a large preference for subunits GluR1 and GluR2 (KD 4-10 nM) over GluR3 and GluR4 (160 600 nM) which implies that tests with brain membranes preferentially reveal drug effects produced at the former two subunits. Lastly, data are shown which highlight the importance of optimizing experimental conditions in filtration assays, for instance by always including thiocyanate in wash buffers. PMID- 16256077 TI - Chronic treatment with amyloid beta(1-42) inhibits non-cholinergic high-affinity choline transport in NG108-15 cells through protein kinase C signaling. AB - We investigated the influence of the amyloid-beta-peptide(1-42) on hemicholinum-3 sensitive high-affinity choline uptake in NG108-15 cells. RT-PCR analysis revealed the presence of mRNA for a choline transporter-like protein but not for cholinergic high-affinity choline transporter. Differentiation of cells increased both hemicholinum-3-sensitive choline uptake and high-affinity hemicholinium-3 binding. This transport was not influenced by tenfold excess of carnitine. Continuous presence of submicromolar concentrations of amyloid-beta-peptide(1-42) during differentiation resulted in a decrease of both choline uptake and hemicholinium-3 binding. These effects were not present when amyloid-beta peptide(1-42) was added 5 min prior to measurements. Neither differentiation nor amyloid-beta-peptide(1-42) treatment changed levels of choline transporter-like protein mRNA. Protein kinase C inhibition by staurosporine or its inactivation by continuous presence of tetradecanoyl phorbol acetate prevented the inhibitory effect of amyloid-beta-peptide(1-42) treatment on choline uptake. Activation of protein kinase C by tetradecanoyl phorbol acetate during measurement had inhibitory effect on choline uptake in control but not amyloid-beta-peptide(1-42) treated cells. The concentration of amyloid-beta-peptide(1-42) maximally effective on hemicholinium-3-sensitive choline uptake had no effect on cell growth, oxidative activity, membrane integrity, number of surface muscarinic receptors, caspase-3 and -8 activities, or uptake of deoxyglucose. Results demonstrate that long-term treatment with non-toxic concentrations of amyloid beta-peptide(1-42) downregulates choline uptake presumably mediated by a choline transporter-like protein through activation of protein kinase C signaling. The decrease of choline uptake may have relevance to the pathogenesis of Alzheimer's disease. PMID- 16256078 TI - Expression of Trk receptors in otolith-related neurons in the vestibular nucleus of rats. AB - The expression of the three Trk receptors (TrkA, TrkB, and TrkC) in otolith related neurons within the vestibular nuclei of adult Sprague-Dawley rats was examined immunohistochemically. Conscious animals were subjected to sinusoidal linear acceleration along either the anterior-posterior (AP) or interaural (IA) axis on the horizontal plane. Neuronal activation was defined by Fos expression in cell nuclei. Control animals, viz labyrinthectomized rats subjected to stimulation and normal rats that remained stationary, showed only a few sporadically scattered Fos-labeled neurons. Among experimental rats, the number of Fos-labeled neurons and their distribution pattern in each vestibular subnucleus in animals stimulated along the antero-posterior axis were similar to those along the interaural axis. No apparent topography was observed among neurons activated along these two directions. Only about one-third of the Trk immunoreactive neurons in the vestibular nucleus expressed Fos. Double-labeled Fos/TrkA, Fos/TrkB and Fos/TrkC neurons constituted 85-98% of the total number of Fos-labeled neurons in vestibular nuclear complex and its subgroups x and y. Our findings suggest that Trk receptors and their cognate neurotrophins in central otolith neurons may contribute to the modulation of gravity-related spatial information during horizontal head movements. PMID- 16256079 TI - Both estrogen and progesterone attenuate edema formation following diffuse traumatic brain injury in rats. AB - Females have reduced brain edema compared to males after experimental brain trauma, although contradictory reports exist as to whether this is due to either estrogen or progesterone. In the present study, we demonstrate in both male and ovariectomized female rats that a single physiological dose of either hormone at 30 min after diffuse traumatic brain injury reduces both blood brain barrier permeability and edema formation. We conclude that both hormones may contribute to reduce edema in females after brain injury. PMID- 16256081 TI - Nocifensive reflex-related on- and off-cells in the pedunculopontine tegmental nucleus, cuneiform nucleus, and lateral dorsal tegmental nucleus. AB - Cholinergic projections from the pedunculopontine tegmental nucleus (PPTg) to the rostral ventromedial medulla (RVM) have been implicated in nociceptive modulation. The goal of this study was to identify neurons with nocifensive reflex-related activity in the mesopontine tegmentum including the PPTg. This study used the same behavioral neurophysiological classification system to identify neurons as has been extensively described in the RVM. Extracellular microelectrode recording was conducted in lightly anesthetized rats. Changes in firing associated with the noxious heat-evoked tail flick reflex were used to classify neurons as "on-cells" (displayed a burst in neuronal activity associated with the reflex), "off-cells" (displayed a pause in activity), and neutral cells (showed no response). Of 188 neurons studied in 23 rats, 77 were classified as on cells, 14 as off-cells, the remainder as neutral cells. Recordings during periods without noxious stimulation found that some of the on- and off-cells displayed spontaneous transitions between active and silent periods termed cell cycling. The distribution of on- and off-cells in the mesopontine tegmentum overlapped and included the cholinergic PPTg and lateral dorsal tegmental nucleus identified by NADPH diaphorase staining, as well as the cuneiform nucleus and periaqueductal gray. The mesopontine tegmentum thus contains nocifensive reflex-related neurons with neurophysiological characteristics similar to those reported in the RVM. Neurons showing reflex-related activity were frequently encountered in the cholinergic PPTg and LDTg. Further studies will be required to determine whether these neurons modulate nociception through a link to the RVM. PMID- 16256080 TI - Antioxidant-like protein 1 is altered in non-pyramidal cells and expressed in astrocytes in the gerbil hippocampal CA1 region after transient forebrain ischemia. AB - In the present study, we observed chronological changes of antioxidant-like protein 1 (AOP-1) in the gerbil hippocampal CA1 region after 5 min of transient forebrain ischemia using immunohistochemistry and western blot. AOP-1 was significantly altered in the CA1 region after transient ischemia. In the sham operated group, AOP-1 immunoreactivity was detected in pyramidal and non pyramidal cells of the CA1 region. At 30 min after ischemic insult, AOP-1 immunoreactivity and protein level was decreased in the CA1 region. At 12 h after ischemic insult, AOP-1 immunoreactivity and protein level was highest in this region. At this time, after ischemia, AOP-1 immunoreactivity in non-pyramidal cells was high compared to the sham-operated group. Based on double immunofluorescence study, AOP-1-immunoreactive neurons were identified as GABAergic, which were stained with GAD or parvalbumin. Thereafter, AOP-1 immunoreactivity and protein levels were decreased time-dependently. From 4 days after ischemic insult, AOP 1 immunoreactivity was generally expressed in astrocytes. Five days after ischemic insult, AOP-1 immunoreactivity and protein level was increased again to 1.4 folds compared to that of the sham-operated group. In brief, AOP-1 immunoreactivity was increased in GABAergic non-pyramidal cells in the hippocampal CA1 region at early time after ischemic insult and was expressed in astrocytes at late time after ischemia. This result suggests that AOP-1 may be important role in homeostasis of GABAergic neurons because these neurons are resistant to ischemic damage. PMID- 16256082 TI - Ethanol substitutes for the discriminative stimulus effects of m chlorophenylpiperazine. AB - Rats were trained to discriminate the 5-HT agonist m-chlorophenylpiperazine (mCPP, 1 mg/kg, i.p.) from saline. Ethanol (0.1 to 1 g/kg, i.p.) partially substituted for the discriminative stimulus effects of 1 mg/kg mCPP. Methysergide (10 mg/kg, i.p.), a 5-HT(1/2) receptor antagonist, blocked the ability of ethanol (1 g/kg) to substitute for mCPP. The largest dose of ethanol markedly reduced response rate. These findings suggest an important role of serotonin receptors in mediating the discriminative stimulus effects of ethanol. PMID- 16256083 TI - Cortical spreading depression (CSD)-induced tolerance to transient focal cerebral ischemia in halothane anesthetized rats is affected by anesthetic level but not ATP-sensitive potassium channels. AB - We investigated the participation of ATP-sensitive potassium (K(ATP)) channels, adenosine A1 receptors, and the effects of different levels of halothane anesthesia in the development of CSD-induced ischemic tolerance. To elicit CSD, 0.5 M KCl was applied for 2 h to the right hemisphere of halothane anesthetized male Wistar rats. The inhalation concentration of halothane during CSD was maintained at 0.5% (n = 8), 1.0% (n = 8), or 2.0% (n = 8). For control animals, saline was applied instead of KCl (n = 8). To inhibit K(ATP) channels or adenosine A1 receptors, glibenclamide (0.1 mg/kg icv; n = 8), 5-hydroxydeconaoate (5-HD; 100 mg/kg ip; n = 12), or 8-Cyclopentyl-1, 3-dipropylxanthine (DPCPX) (1.0 mg/kg ip; n = 8) was applied before preconditioning during 1.0% halothane anesthesia. Temporary occlusion (120 min) of the right middle cerebral artery was induced 4 days after preconditioning and the infarct volume was measured. Preconditioning elicited under 1.0% halothane reduced cortical infarct volume from 277 +/- 15 mm3 in the control group to 159 +/- 14 mm3 in the CSD group (mean +/- SEM, P < 0.05). In contrast, CSD induced during inhalation of 0.5% or 2.0% halothane did not confer ischemic tolerance. The reduction in infarct area with CSD during inhalation of 1% halothane was not changed in animals treated with glibenclamide or 5-HD or DPCPX. These results uncover a crucial role of halothane level but not of K(ATP) channels or adenosine A1 receptors in the preconditioning effects of CSD. PMID- 16256086 TI - Metabotropic glutamate receptor agonists modify the pyloric output of the crustacean stomatogastric ganglion. AB - We have studied the effects of groups I, II, and III metabotropic glutamate receptor (mGluR) agonists and antagonists on pyloric activity in the stomatogastric ganglion (STG) of the Caribbean spiny lobster Panulirus argus. We have found that agonists for all three groups of mGluRs modify the pyloric output. The group I agonist, l-quisqualic acid (l-QA), activated the pyloric central pattern generator (CPG). When the pyloric rhythm was partially suppressed by sucrose-block of input fibers in the stomatogastric nerve (stn), l-QA accelerated the rhythmic activity. In addition, the number of spike discharges was increased in pyloric motoneurons: pyloric (PY), and lateral pyloric (LP). In completely blocked preparations, a slow pyloric rhythm was initiated by l-QA. Groups II and III agonists exerted an inhibitory effect on pyloric activity. The group II agonist, (2S,1'S,2'S)-2-(Carboxycyclopropyl)glycine (L-CCG-I), decreased both the frequency of the pyloric rhythm and the number of spike discharges in the motoneurons: ventricular dilator (VD), PY, and LP. The effects of L-CCG-I were dose-dependent. The group III agonist, l-(+)-2-Amino-4-phosphonobutyric acid (l-AP4), slightly decreased the frequency of the pyloric rhythm and suppressed spike discharges in the VD neuron. All effects of mGluR agonists were reversible. The effect of l-QA was blocked by the broad spectrum mGluR antagonist (S)-Methyl 4-carboxyphenylglycine (MCPG). The inhibitory effect of L-CCG-I was prevented by MCPG and by the group II/III mGluR antagonist (RS)-alpha-Methyl-4 phosphonophenylglycine (MPPG), and was partially blocked by the group II mGluR antagonist (RS)-1-amino-5-phosphonoindan-1-carboxylic acid (APICA). The inhibitory effect of l-AP4 was blocked by MPPG and partially blocked by APICA. PMID- 16256087 TI - Circadian variation and intermuscular correlation of rabbit jaw muscle activity. AB - The activation of jaw muscles varies with different tasks and must be coordinated to ensure proper function of the masticatory system. The activation patterns might differ in various muscles or over the time course. In order to evaluate the activation patterns and the intermuscular correlation during normal daily activity the electromyograms (EMG) of the superficial and deep masseter, medial pterygoid and digastric muscles were continuously recorded in rabbits and related to activity levels. Muscle use was assessed as the relative time per hour (duty time) during which predefined levels of the peak-EMG of the day were exceeded. Pearson's correlation of duty times was calculated for 6 muscle pairs at various activity levels. The duty times of the muscles differed significantly at levels exceeding 50% of the peak-EMG. The animals exhibited apparent intraday variations of duty times revealing a circadian covariant pattern of muscle use. These variations, however, were different in each individual animal. The activation of pairs of jaw-closing muscles was more highly correlated than that of pairs consisting of a jaw-closing and a jaw-opening muscle. The mutual dependence of hourly muscle activity among jaw-closers and among jaw-closers and jaw-openers varied with the activity level suggesting that those muscle groups might be independently controlled during non-powerful and powerful motor behaviors. PMID- 16256084 TI - A detailed characterization of loud noise stress: Intensity analysis of hypothalamo-pituitary-adrenocortical axis and brain activation. AB - The present studies were undertaken to help determine the putative neural circuits mediating activation of the hypothalamo-pituitary-adrenocortical (HPA) axis and the release of adrenocorticotropin hormone (ACTH) and corticosterone in response to the perceived threat of loud noise. This experiment involved placing rats in acoustic chambers overnight to avoid any handling and context changes prior to noise exposure, which was done for 30 min (between 9:00 and 10:00 am) at intensities of 80, 85, 90, 95, 100, 105, and 110 dBA in different groups (n = 8), and included a background condition (60 dBA ambient noise). This manipulation produced a noise-intensity-related increase in plasma ACTH and corticosterone levels, with levels beginning to rise at approximately 85 dBA. c-fos mRNA induction was very low in the brains of the control and 80 dBA groups, but several brain regions displayed a noise-intensity-related induction. Of these, several forebrain regions displayed c-fos mRNA induction highly correlated (r > 0.70) with that observed in the paraventricular hypothalamic nucleus and plasma ACTH levels. These regions included the ventrolateral septum, the anteroventral subiculum, several preoptic nuclei, the anterior bed nucleus of the stria terminalis (BNST), the anterior paraventricular nucleus of the thalamus, and the medial subdivision of the medial geniculate body. Together with prior findings with audiogenic stress, the present results suggest that either or both the anterior BNST or the lateral septum is ideally situated to trigger HPA axis activation by stimuli that are potentially threatening. PMID- 16256085 TI - 5-HT1B receptors modulate the feeding inhibitory effects of enterostatin. AB - Serotonin (5-HT) is considered to play an important role in control of appetite. Enterostatin has been shown to alter 5-HT release in the brain, and non-specific 5-HT antagonists blocked the anorectic response to icv enterostatin. The aim of this study was to further identify which 5-HT receptor subtype mediates the enterostatin feeding behavior and whether this effect occurs due to action in the PVN. Wild-type and 5-HT2C receptor-/- (KO) mice and normal Sprague-Dawley rats were used in these experiments. All animals were fed a high fat diet. Enterostatin (120 nmol, i.p.) reduced the intake of high fat diet in 5-HT2C receptor mutant mice (saline 4.54 +/- 0.47 kcal vs. Ent 2.53 +/- 0.76 kcal) 1 h after injection. A selective 5-HT1B antagonist (GR55526, 40 mg/kg body weight, i.p.) blocked the enterostatin hypophagic effects in these KO mice. Rats were implanted with cannulas into the amygdala and the ipsilateral PVN. The 5-HT receptor antagonists metergoline (non-specific receptor subtypes 1 and 2), or ritanserin (selective 2C), or GR55562 (selective l B) was injected into the PVN prior to enterostatin (0.01 nmol) injection into the amygdala. Enterostatin reduced food intake (saline: 5.80 +/- 0.59 g vs. enterostatin 3.47 +/- 0.56 g, P < 0.05 at l h). Pretreatment with either metergoline (10 nmol) or GR55526 (10 nmol) but not ritanserin (10 nmol) into the PVN attenuated the anorectic response to amygdala enterostatin. The data imply that the enterostatin anorectic response may be modulated by 5-HT1B receptors and that a neuronal pathway from the amygdala to the PVN regulates the enterostatin response through activation of 5 HTlB receptors in PVN. PMID- 16256088 TI - Selective upregulation of RB3/stathmin4 by ciliary neurotrophic factor following optic nerve axotomy. AB - In this study, we examined the cellular responses of stathmin-related proteins in the rat retina following optic nerve (ON) axotomy. To examine the distribution of stathmin-related gene products, we performed semi-quantitative reverse transcription polymerase chain reaction (RT-PCR), in situ hybridization (ISH) and immunohistochemical analyses. Retrograde labeling using a fluorescein tracer, fluorogold (FG), was used for the identification of retinal ganglion cells (RGCs). RT-PCR and ISH analyses indicated that the expression of RB3 was specifically increased in the ganglion cell layer (GCL) comparing to other members of stathmin-related gene family examined 3 days following the ON axotomy. When brain-derived neurotrophic factor was administrated intravitreously, the induction of RB3 mRNA sustained up to 7 days after axotomy, although the peak induction level was unchanged. In contrast, ciliary neurotrophic factor (CNTF) administration increased the peak level of RB3 mRNA induction significantly at 3 days after axotomy. Immunohistochemistry in combination with the retrograde labeling of axotomized cells by FG revealed that RB3 was increased following axotomy in FG-labeled RGCs. These data suggest that RB3 is the unique response protein in the stathmin-related proteins following ON axotomy and the induced RB3 may play a critical role in the CNTF-induced response on the axotomized RGCs, e.g. axonal regeneration and/or neuroprotection. PMID- 16256089 TI - Do we need new antiarrhythmic compounds in the era of implantable cardiac devices and percutaneous ablation? PMID- 16256090 TI - Oral administration of pyridostigmine bromide and huperzine A protects human whole blood cholinesterases from ex vivo exposure to soman. AB - Cholinesterases (ChEs) are classified as acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) according to their substrate specificity and sensitivity to selected inhibitors. The activities of AChE in red blood cells (RBC-AChE) and BChE in serum can be used as potential biomarkers of suppressed and/or heightened activity in the central and peripheral nervous systems. Exposure to organophosphate (OP) chemical warfare agents (CWAs), pesticides, anesthetics, and a variety of drugs such as cocaine, as well as some neurodegenerative and liver disease states, selectively reduces AChE or BChE activity. In humans, the toxicity of pesticides is well documented. Therefore, blood cholinesterase activity can be exploited as a tool for confirming exposure to these agents and possible treatments. Current assays for measurement of RBC AChE and serum BChE require several labor-intensive processing steps, suffer from wide statistical variation, and there is no inter-laboratory conversion between methods. These methods, which determine only the serum BChE or RBC-AChE but not both, include the Ellman, radiometric, and deltapH (modified Michel) methods. In contrast, the Walter Reed Army Institute of Research Whole Blood (WRAIR WB, US Patent #6,746,850) cholinesterase assay rapidly determines the activity of both AChE and BChE in unprocessed (uncentrifuged) whole blood, uses a minimally invasive blood sampling technique (e.g., blood from a finger prick), and is semi automated for high-throughput using the Biomek 2000 robotic system. To date, the WRAIR whole blood assay was used to measure AChE and BChE activities in human blood from volunteers in FDA clinical trials. In the first FDA study, 24 human subjects were given either 30 mg PB orally (n = 19) or placebo (n = 5). Blood samples were obtained pre-dosing and 2.5, 5, 8, and 24 h post-dosing. The samples were analyzed for AChE and BChE activity using the WRAIR WB robotic system, and for PB concentration by HPLC. We found that maximal inhibition of AChE (26.2%) and concentration of PB (17.1 ng/mL) occurred at 2.5 h post-PB dosing. AChE activity returned to almost 100% of pre-dose values by 6 h. A dose-dependent linear correlation was found between the amount of PB measured in the blood and the inhibition of AChE. Following soman (GD) exposure, recovered AChE activity was similar to levels that were reversibly protected by the PB administration. Therefore, the WRAIR ChE WB data clearly supports the conclusion that PB is an effective pre-treatment drug for nerve agent exposure (GD). In the second FDA human study for the treatment of Alzheimer's disease, the WRAIR ChE WB assay was used to determine the RBC-AChE and serum BChE profile of healthy elderly volunteers receiving Huperzine A. Huperzine A is a plant-derived reversible and selective AChE inhibitor compared to BChE, and is a more potent inhibitor of AChE than PB. Huperzine A is available as a nutraceutical, a natural supplement reported to improve memory, and has a variety of neuroprotective effects. Individuals received an increasing dose regimen of huperzine A (final dose 200 microg after 4 weeks), which produced more than 50% inhibition of RBC-AChE. Huperzine A was well tolerated by these patients at doses that sequestered more RBC-AChE than PB, and thus warrants further study as a prophylaxis for OP poisoning in addition to Alzheimer's therapy. Due to the documented use of OPs by terrorists and in warfare around the globe, Federal, State, and local authorities need a reliable, fast, inexpensive, and standard method for confirming such an assault in order to initiate appropriate containment, decontamination, and treatment measures. This assay is ideal for prescreening military personnel for atypical ChE activities that would preclude their deployment to areas of potential CWA exposure. The WRAIR WB ChE assay will fulfill the requirement for rapid and reliable monitoring of such exposure in military and civilian populations. PMID- 16256091 TI - Expression and distribution of acetylcholinesterase among the cellular components of the neuromuscular junction formed in human myotube in vitro. AB - The results of our recent investigations on the expression and distribution of acetylcholinesterase (EC. 3.1.1.7, AChE) in the experimental model of the in vitro innervated human muscle are summarized and discussed here. This is the only model allowing studies on AChE expression at all stages of the neuromuscular junction (NMJ) formation in the human muscle. Since it consists not only of the motor neurons and myotubes but also of glial cells, which are essential for the normal development of the motor neurons, NMJs become functional and differentiated in this system. We followed AChE expression at various stages of the NMJ formation and in the context of other events characteristic for this process. Neuronal and muscular part were analysed at both, mRNA and mature enzyme level. AChE is expressed in motor neurons and skeletal muscle at the earliest stages of their development, long before NMJ starts to form and AChE begins to act as a cholinergic component. Temporal pattern of AChE mRNA expression in motor neurons is similar to the pattern of mRNA encoding synaptogenetic variant of agrin. There are no AChE accummulations at the NMJ at the early stage of its formation, when immature clusters of nicotinic receptors are formed at the neuromuscular contacts and when occasional NMJ-mediated contractions are already observed. The transformation from immature, bouton-like neuromuscular contacts into differentiated NMJs with mature, compact receptor clusters, myonuclear accumulations and dense AChE patches begins at the time when basal lamina starts to form in the synaptic cleft. Our observations support the concept that basal lamina formation is the essential event in the transformation of immature neuromuscular contact into differentiated NMJ, with the accumulation of not only muscular but also neuronal AChE in the synaptic cleft. PMID- 16256092 TI - Proceedings of the VIII International Meeting on Cholinesterases, September 26 30, 2004, Perugia, Italy. PMID- 16256093 TI - In vivo cholinesterase inhibitory specificity of organophosphorus nerve agents. AB - The purpose of this project was to determine and compare the time-related changes in blood, brain, and tissue acetylcholinesterase (AChE) activity during the first hour after exposure to six organophosphorus nerve agents (GA, GB, GD, GF, VR, and VX) in Hartley guinea pigs. Animals were pretreated with atropine methyl nitrate (1.0mg/kg, i.m.) to minimize peripheral toxic effects 15 min before they were given a 1.0 x LD50 subcutaneous dose of a nerve agent. At 0, 5, 10, 15, 30, and 60 min after nerve agent, animals were humanely euthanized. Blood was collected and brain regions (brainstem, cortex, hippocampus, midbrain, cerebellum, striatum, and spinal cord) and peripheral tissues (diaphragm, skeletal muscle, and heart) were dissected and processed for AChE activity. All six nerve agents produced maximum inhibition of AChE in red blood cells between 5 and 10% of the control within 10 min after exposure. In whole blood, differential effects were observed among the agents: GB, GD, and GF produced more rapid and greater inhibition than did GA, VR, and VX. GF was the most rapid, producing a maximum inhibition to 5% of the control in 5 min, while VR and VX were slower reaching maximum inhibition to 30% of the control at 15 min. The enzyme activity in the majority of the brain regions was more markedly inhibited by the G-agents than by the V-agents. The G-agents caused rapid AChE inhibition, reaching maximum levels (20-30% of control) at 15 min and GA produced the most rapid effects. V-agents produced much slower and less AChE inhibition, reaching maximum (35-60% of control) at 30 min. In the diaphragm, VR, VX, and GD produced more rapid and greater AChE inhibition than other G-agents; GA produced the slowest and least inhibition. In the skeletal muscle, VX induced the most rapid and severe inhibition, while GA the least inhibition. In the heart, all agents produced very rapid inhibition, and GD produced the most severe inhibition of AChE activity. These observations suggest that G-agents and V-agents are tissue compartment specific in their ability to inhibit AChE activity. PMID- 16256094 TI - Rational polynomial equation helps to select among homeomorphic kinetic models for cholinesterase reaction mechanism. AB - The hydrolysis of substrates by cholinesterases does not follow the Michaelis Menten reaction mechanism. In addition to the inhibition by excess substrate, these enzymes often show an unexpectedly high activity at low substrate concentrations. It seems that these deviations are the consequence of an unusual architecture of the active site, buried deep inside the core of the molecule. Kinetic data and structural evidence allow for a detailed prediction of the events during a very fast substrate turnover. Recently, we presented a procedure which provides an unbiased framework for mathematical modelling of the complex cholinesterase reaction [J. Stojan, M. Golicnik, D. Fournier, Rational polynomial equation as an unbiased approach for the kinetic studies of Drosophila melanogaster acetylcholinesterase reaction mechanism, Biochim. Biophys. Acta 1703 (2004) 53-61]. It is based on regression analysis of a rational polynomial using classical initial rate data. Here, we extend the use of the rational polynomial rate equation for finding and comparing some selected homeomorphic reaction schemes useful for the mechanistic interpretation of cholinesterase kinetic data. PMID- 16256095 TI - Regio- and stereoselectivity in preparation of 2-deoxysugar glycoglycerolipid derivatives evaluated by high resolution 1H NMR. AB - New glycoglycerolipids, derivatives of 2-deoxysugars bearing one or two fatty acid chains have been synthesised. Various levels of regio- and stereoselectivity have been attained for the triphenylphosphine hydrobromide (TPHB) catalysed addition of the glycerol moiety to some representative glycals. The influence of the structure of glycal derivatives in glycosylation reactions is discussed. PMID- 16256096 TI - Micellization of conjugated chenodeoxy- and ursodeoxycholates and solubilization of cholesterol into their micelles: comparison with other four conjugated bile salts species. AB - Micelle formations of sodium glyco- and taurochenodeoxycholate (NaGCDC and NaTCDC) and sodium glyco- and tauroursodeoxycholates (NaGUDC and NaTUDC) was studied at 308.2 K for their critical micelle concentrations at various NaCl concentrations by pyrene fluorescence probe, and the degree of counterion binding to micelle was determined using the Corrin-Harkins plots. The degree of counterion binding was found to be 0.37-0.38 for chenodeoxycholate conjugates, while the determination of the degree was quite difficult for ursodeoxycholate conjugates. The change of I1/I3 values on the fluorescence spectrum with the conjugate bile salt concentration suggested two steps for their bile salt aggregation. The first step is a commencement of smaller aggregates, the first cmc, and the second one is a starting of stable aggregates, the second cmc. The aggregation number was determined at 308.2 K and 0.15 M NaCl concentration by static light scattering: 16.3 and 11.9 for sodium NaGCDC and NaTCDC, and 7.9 and 7.1 for NaGUDC and NaTUDC, respectively. The solubilization of cholesterol into the bile salt micelles in the presence of coexisting cholesterol phase and the maximum additive concentration (MAC) of cholesterol was determined against the bile salt concentration. The standard Gibbs energy change for the solubilization was evaluated, where the micelles were regarded as a chemical species. The solubilization was stabilized in the order of NaGUDC approximately = NaTUDC < NaTC < NaGC < NaTCDC < NaGCDC < NaTDC < NaGDC, where the preceding results were taken into the order. PMID- 16256097 TI - Serum vitamin C concentration is influenced by haptoglobin polymorphism and iron status in Chinese. AB - BACKGROUND: Vitamin C is a powerful antioxidant (free radical scavenger). Apart from the diet, other factors regulating its catabolism may affect its serum concentration. Haptoglobin (Hp) is a plasma protein participating in iron metabolism. It shows a genetic polymorphism which shows marked geographical differences. We investigated the relationship between vitamin C, iron status and haptoglobin polymorphism in Chinese men and women. METHODS: Iron status markers were compared according to Hp phenotypes determined by chemiluminescence detection in 110 healthy Chinese subjects. The concentration of haptoglobin was determined using an immunoturbidimetric method. Serum vitamin C was tested by a 2,4-dinitrophenylhydrazine based method. RESULTS: In Chinese, the haptoglobin phenotype distribution was 10.0% Hp 1-1, 33.6% Hp 2-1, and 56.4% Hp 2-2. In the study group, serum vitamin C concentration was associated with haptoglobin type, showing lowest values in serum from Hp 2-2 subjects in males (p=0.028, ANOVA). In contrast to Hp phenotype, Hp concentration did not affect vitamin C concentration. Hp 2-2 shows higher haptoglobin (p=0.002 (ANOVA)) than individuals with the other types. Furthermore, vitamin C was influenced by (log)ferritin levels. In Chinese, vitamin C is influenced by haptoglobin polymorphism and iron status. CONCLUSION: The present findings support the role of non-nutritional factors in vitamin C status. PMID- 16256098 TI - Hypercoagulability markers in young asymptomatic heterozygous carriers of factor V Leiden (G1691A) or prothrombin (G20210A) variant. AB - BACKGROUND: Mutations in factor V (factor V Leiden-G1691A) and prothrombin (G20210A) genes are important risk factors for thrombophilia due to their high incidence in patients with thromboembolic events, especially among the young. However, it is not clear if levels of hypercoagulability markers are significantly altered in asymptomatic young carriers of factor V Leiden or prothrombin G20210A. METHODS: Hemostatic status of 32 asymptomatic young individuals carrying these mutations and of 18 normal control individuals was investigated through the determination of plasma thrombomodulin (TM), prothrombin fragment 1+2 (F1+2), thrombin-antithrombin complex (TAT) and D-dimer. RESULTS: No significant differences were observed in these hemostatic markers when comparing groups of individuals carrying mutations and the control group. CONCLUSION: Analysis of these results leads to the conclusion that the presence of these mutations, in the absence of acquired risk factors, does not constantly predispose these young carriers to a state of hypercoagulability. PMID- 16256099 TI - Fgf19 regulated by Hh signaling is required for zebrafish forebrain development. AB - Fibroblast growth factor (Fgf) signaling plays important roles in brain development. Fgf3 and Fgf8 are crucial for the formation of the forebrain and hindbrain. Fgf8 is also required for the midbrain to form. Here, we identified zebrafish Fgf19 and examined its roles in brain development by knocking down Fgf19 function. We found that Fgf19 expressed in the forebrain, midbrain and hindbrain was involved in cell proliferation and cell survival during embryonic brain development. Fgf19 was also essential for development of the ventral telencephalon and diencephalon. Regional specification is linked to cell type specification. Fgf19 was also essential for the specification of gamma aminobutyric acid (GABA)ergic interneurons and oligodendrocytes generated in the ventral telencephalon and diencephalon. The cross talk between Fgf and Hh signaling is critical for brain development. In the forebrain, Fgf19 expression was down-regulated on inhibition of Hh but not of Fgf3/Fgf8, and overexpression of Fgf19 rescued partially the phenotype on inhibition of Hh. The present findings indicate that Fgf19 signaling is crucial for forebrain development by interacting with Hh and provide new insights into the roles of Fgf signaling in brain development. PMID- 16256100 TI - The effect of agrin and laminin on acetylcholine receptor dynamics in vitro. AB - Using optical imaging assays, we investigated the dynamics of acetylcholine receptors (AChRs) at laminin-associated clusters on cultured myotubes in the absence or presence of the nerve-derived clustering factor, agrin. Using fluorescence recovery after photobleaching (FRAP) on fluorescent bungarotoxin labeled receptors, we found that approximately 9% of original fluorescence was recovered after 8 h as surface AChRs were recruited into clusters. By quantifying the loss of labeled receptors and the recovery of fluorescence after photobleaching, we estimated that the half-life of clustered receptors was approximately 4.5 h. Despite the rapid removal of receptors, the accumulation of new receptors at clusters was robust enough to maintain receptor density over time. We also found that the AChR half-life was not affected by agrin despite its role in inducing the aggregation of AChRs. Interestingly, when agrin was added to myotubes grown on laminin-coated substrates, most new receptors were not directed into preexisting laminin-induced clusters but instead formed numerous small aggregates on the entire muscle surface. Time-lapse imaging revealed that the agrin-induced clusters could be seen as early as 1 h, and agrin treatment resulted in the complete dissipation of laminin-associated clusters by 24 h. These results reveal that while laminin and agrin are involved in the clustering of receptors they are not critical to the regulation of receptor metabolic stability at these clusters, and further argue that agrin is able to rapidly and fully negate the laminin substrate clustering effect while inducing the rapid formation of new clusters. PMID- 16256101 TI - No-threshold dose-response curves for nongenotoxic chemicals: findings and applications for risk assessment. AB - We tested the hypothesis that no threshold exists when estradiol acts through the same mechanism as an active endogenous estrogen. A Michaelis-Menten (MM) equation accounting for response saturation, background effects, and endogenous estrogen level fit a turtle sex-reversal data set with no threshold and estimated the endogenous dose. Additionally, 31 diverse literature dose-response data sets were analyzed by adding a term for nonhormonal background; good fits were obtained but endogenous dose estimations were not significant due to low resolving power. No thresholds were observed. Data sets were plotted using a normalized MM equation; all 178 data points were accommodated on a single graph. Response rates from approximately 1% to >95% were well fit. The findings contradict the threshold assumption and low-dose safety. Calculating risk and assuming additivity of effects from multiple chemicals acting through the same mechanism rather than assuming a safe dose for nonthresholded curves is appropriate. PMID- 16256102 TI - Effects of glucocorticoid receptor antagonists on allodynia and hyperalgesia in mouse model of neuropathic pain. AB - Injury to the spinal nerves of mice induces mechanical allodynia and thermal hyperalgesia. In the injured spinal cord, the expression of glucocorticoid receptor mRNA was increased, whereas it was decreased in N-type Ca(2+)-channel deficient mice, in which neuropathic pain is eliminated. Intrathecal and intraperitoneal injection of the glucocorticoid receptor antagonist RU486 produced antinociceptive effects, whereas intracerebroventricular injection was without effect. The more selective antagonist dexamethasone 21-mesylate suppressed both mechanical allodynia and thermal hyperalgesia. These results suggest that spinal glucocorticoid receptors play an important role in neuropathic pain, and that controlling the activity of glucocorticoid receptors may be of great importance in the treatment of neuropathic pain. PMID- 16256103 TI - Effects of mometasone furoate on a rat allergic rhinitis model. AB - The present study was undertaken to clarify the effects of mometasone on nasal symptoms induced by repeated intranasal application of antigen in sensitized rats in comparison with that of chlorpheniramine. Rats received mometasone intranasally or chlorpheniramine orally 1 h before a topical antigen challenge for 7 days. Mometasone caused a decrease in the instances of nasal rubbing and an inhibition of this response was observed during the treatment period. Almost identical findings were observed with chlorpheniramine. This response was inhibited, even after the interruption of mometasone treatment, while such an effect was not observed with chlorpheniramine. On day 36, the changes in sensitivity to histamine were investigated. Unlike chlorpheniramine, hypersensitivity to histamine was significantly reduced in the mometasone-treated group. The passive cutaneous anaphylaxis titers were elevated and reached a maximum 8 days after the start of the topical antigen challenge. The passive cutaneous anaphylaxis titer in the mometasone-treated group was significantly lower than that in the control group. The results indicated that mometasone is effective in allergic rhinitis, not only during the period of application, but also after the interruption of application. PMID- 16256104 TI - Juvenile pig detrusor: effects of propiverine and three of its metabolites. AB - In isolated detrusor strips, propiverine is known to be effective to decrease contractions elicited by electric field stimulation (EFS). Here we investigated whether the metabolites M-5, M-6 and M-14 of propiverine retain the pharmacological properties of the parent compound also in juvenile organisms. EFS induced contractions of detrusor strips from juvenile pigs are more sensitive to atropine than strips from mature pigs. The atropine-resistant component of contraction is also significantly larger in juvenile pigs. Propiverine, its metabolites M-5, M-14 and also tolterodine completely reduced detrusor contraction in juvenile pigs. M-6 almost did not affect atropine-resistant contractions. We conclude that juvenile pig detrusors possess a higher atropine resistant component of EFS-elicited contraction. Nevertheless order of potency and efficacy of propiverine and its metabolites M-5 and M-14 are similar in juvenile and mature pigs, while M-6 only reduces atropine-sensitive contractions in the juvenile organism. PMID- 16256105 TI - The role of dopamine in human addiction: from reward to motivated attention. AB - There is general consensus among preclinical researchers that dopamine plays an important role in the development and persistence of addiction. However, the precise role of dopamine in addictive behaviors is far from clear and only a few clinical studies on the role of dopamine in human addiction have been conducted so far. The present paper reviews studies addressing the role of dopamine in humans. There is substantial and consistent evidence that dopamine is involved in the experience of drug reward in humans. Dopamine may also be involved in motivational processes such as drug craving. However, given the inconsistent findings of studies using dopamine receptor (ant)agonists, the role of dopamine in the experience of craving is far from resolved. Recent theories claiming that dopamine signals salience and makes the brain paying attention to biological relevant stimuli may provide an interesting framework for explaining addictive behaviors. There is accumulating evidence that patients with drug and alcohol addiction have an aberrant focus on drug-related stimuli. Although there is some preliminary support for the role of dopamine in these attention processes, more studies have to be carried out in order to test the validity of these theories in human subjects. PMID- 16256106 TI - Antinociceptive effect of oxycodone in diabetic mice. AB - The effect of oxycodone on thermal hyperalgesia in streptozotocin-induced diabetic mice was examined. The antinociceptive response was assessed by recording the latency in the tail-flick test using the radiant heat from a 50-W projection bulb on the tail. The tail-flick latency in diabetic mice was significantly shorter than that in non-diabetic mice. When diabetic mice were treated with oxycodone (5 mg/kg, s.c.), the tail-flick latency in diabetic mice was prolonged to the level considerably longer than the baseline latencies of non diabetic mice. However, s.c. administration of morphine (5 mg/kg) did not produce a significant inhibition of the tail-flick response in diabetic mice. Oxycodone, at doses of 1.25-5.0 mg/kg administered s.c., produced a dose-dependent increase in the tail-flick latencies in both diabetic and non-diabetic mice. The antinociceptive effect of oxycodone was antagonized by pretreatment with a selective delta-opioid receptor antagonist, beta-funaltrexamine (20 mg/kg, s.c.), in both non-diabetic and diabetic mice. In non-diabetic mice, pretreatment with a selective kappa-opioid receptor antagonist, nor-binaltorphimine (20 mg/kg, s.c.) had no effect on the peak antinociceptive effect of oxycodone observed 30 min after administration, however, it slightly but significantly reduced oxycodone induced antinociception observed 60 and 90 min after administration. On the other hand, pretreatment with nor-binaltorphimine practically abolished the peak (30 min) and persistent (60 and 90 min) antinociceptive effects of oxycodone in diabetic mice. Naltrindole (35 mg/kg, s.c.), a selective delta-opioid receptor antagonist, had no effects on the antinociceptive effect of oxycodone in both non diabetic and diabetic mice. These results suggest that the antinociceptive effects of oxycodone may be mediated by mu- and kappa-opioid receptors in diabetic mice, whereas it may interact primarily with mu-opioid receptors in non diabetic mice. PMID- 16256107 TI - Role of the renal nerves in gamma-aminobutyric acid-induced antihypertensive effect in spontaneously hypertensive rats. AB - The aim of this study was to clarify the role of the renal sympathetic nerves in the gamma-aminobutyric acid (GABA)-induced hypotensive effect in spontaneously hypertensive rats. Male spontaneously hypertensive rats (SHR/Izm) aged 7 weeks were divided into four groups on the basis of diet (containing 0.05% GABA, or GABA-free control diet) and operation (renal sympathetic-denervated or sham operated) (n = 10, each). Water intake, urine volume and urinary sodium were, or tended to be, slightly higher, while plasma renin activity was significantly lower in the GABA group than the GABA-free control group. GABA inhibited the development of hypertension in sham-operated spontaneously hypertensive rats but not in renal-denervated spontaneously hypertensive rats. Plasma renin activity was significantly higher in sham-operated spontaneously hypertensive rats fed the control diet than in the other three groups. These results suggest that a reduction in the effects induced by the renal nerves may play an important role in the hypotensive effect induced in spontaneously hypertensive rats by chronic dietary administration of GABA. PMID- 16256108 TI - TSC-36/FRP inhibits vascular smooth muscle cell proliferation and migration. AB - OBJECTIVE: In-stent restenosis is a vascular proliferation/migration disorder characterized by hyperplasia of vascular smooth muscle cells (VSMCs). Because mounting evidence suggests that the therapeutic potential of anti-proliferation and anti-migration therapy, we investigated possible inhibitory effects of the matricellular protein TGF-beta-stimulated clone 36 (TSC-36) on vascular smooth muscle cell proliferation and migration in vitro and in vivo. METHODS: Human umbilical artery smooth muscle cells (SMCs) were treated with inducting agents daidzein or estradiol. TSC-36 expression was detected by nested competitive PCR and in situ hybridization. TSC-36 was expressed in Origami (DE3) cells. The recombinant protein was used to immunize rabbits to produce polyclonal antibodies. VSMCs were treated with various concentrations of recombinant TSC-36 (rTSC-36) protein and daidzein. The MTT assay was used to analyze for cell proliferation. A transwell system was used to detect cell migration. Flow cytometry was used to detect cell phase. A rat carotid artery balloon injury model was duplicated. The rats were treated with daidzein or solvent control. Animals were sacrificed 5 weeks later, and injured arteries were taken for pathology and histology. RESULTS: TSC-36 mRNA and protein expression was induced in SMCs. Cell proliferation and migration were inhibited by rTSC-36. rTSC-36 caused accumulation of SMCs in G2 phase. The inducting agent daidzein decreased neo-intima proliferation. TSC-36 mRNA and protein expression was induced and expressed in the neo-intima. CONCLUSION: TSC-36 can be induced in VSMCs and inhibits VSMCs proliferation in vitro and in vivo. PMID- 16256109 TI - Babesia canis vogeli: a novel PCR for its detection in dogs in Australia. AB - Babesia canis vogeli is known to cause disease in dogs in Australia, and the rapid detection of various subspecies would enable effective treatment and management. A 21 bp oligonucleotide, "Bab-f" was proposed for the production of larger PCR products with high species specificity that would enable effective sequence analyses to yield subspecies identification. The new forward primer when paired with a previously reported "Babesia common" reverse primer generated a 394 bp product which was successfully amplified and provided subspecies differentiation by sequence analyses. Specificity and sensitivity were reported at 100% on a cohort of 55 dogs. PMID- 16256110 TI - Energy allocation patterns in Biomphalaria alexandrina snails in response to cadmium exposure and Schistosoma mansoni infection. AB - This study was aimed to investigate the effects of both parasitism and environmental stress on the growth, reproduction, and survival of Biomphalaria alexandrina snails. Resource allocation strategies may be influenced by both biotic and abiotic factors. Using the planorbid snail B. alexandrina and Schistosoma mansoni, this hypothesis was examined by raising snails fed the same diet under two stressors (infection and Cd exposure). The snails divided into four groups, uninfected, infected, Cd-exposed uninfected, and Cd-exposed infected snails. Egg production, growth, and survival of the snails were monitored over a 9-week period postinfection. Inhibition of snail reproductive activity by parasitism results in increased snail growth in the first week postinfection, termed gigantism, during which the snail is hypothesized to allocate excess energy normally used for reproduction to somatic growth. Infection status and Cd exposure had significant effects on snail growth and reproduction. The infected and Cd-exposed infected snails exhibiting reduced survival relative to snails of other treatments. It was found that parasite development influenced by Cd exposure. Results of this study suggest that energy allocation patterns are context-dependent in B. alexandrina snails, influenced by infection and Cd exposure. PMID- 16256111 TI - Co-exposure to benzo[a]pyrene and UVA induces phosphorylation of histone H2AX. AB - Phosphorylation of histone H2AX (termed gamma-H2AX) was recently identified as an early event after induction of DNA double strand breaks (DSBs). We have previously shown that co-exposure to benzo[a]pyrene (BaP), a wide-spread environmental carcinogen, and ultraviolet A (UVA), a major component of solar UV radiation, induced DSBs in mammalian cells. In the present study, we examined whether co-exposure to BaP and UVA generates gamma-H2AX in CHO-K1 cells. Single treatment with BaP (10(-9)-10(-7)M) or UVA ( approximately 2.4 J/cm(2)) did not result in gamma-H2AX, however, co-exposure drastically induced foci of gamma-H2AX in a dose-dependent manner. gamma-H2AX could be detected even at very low concentration of BaP (10(-9)M) plus UVA (0.6J/cm(2)), which did not change cell survival rates. NaN(3) effectively inhibited the formation of gamma-H2AX induced by co-exposure, indicating the contribution of singlet oxygen. This is the first evidence that co-exposure to BaP and UVA induced DSBs, involving gamma-H2AX. PMID- 16256112 TI - Stg 1 is a novel SM22/transgelin-like actin-modulating protein in fission yeast. AB - We identified a novel actin-modulating protein Stg 1 in the fission yeast Schizosaccharomyces pombe. Stg 1 is similar to mammalian SM22/transgelin, and biochemical experiments showed that Stg 1 crosslinked F-actin. Microscopic observation suggested that Stg 1 was a component of actin patch. Overexpression of Stg 1 caused a defect in cytokinesis by suppressing the formation of a contractile ring and formation of abnormal aggregates of F-actin in the ends and mid-region of cells. Although distribution of the actin cytoskeleton was not affected by disrupting Stg 1(+), genetic interaction suggested that Stg 1 was likely involved in controlling the organization of the actin cytoskeleton in cell morphogenesis and cytokinesis in fission yeast. PMID- 16256113 TI - Synthesis of (di)nucleoside polyphosphates by the ubiquitin activating enzyme E1. AB - Previous work from this laboratory had shown that ligases may catalyze the synthesis of (di)nucleoside polyphosphates. Here, we show that one of the enzymes of the proteasome system (E1 or the ubiquitin (Ub) activating enzyme, EC 6.3.2.19) catalyzes very effectively (k(cat) = 0.29+/-0.05 s(-1)) the transfer of AMP from the E-AMP-ubiquitin complex to tripolyphosphate or tetrapolyphosphate with formation of adenosine tetra- or pentaphosphate (p4A or p5A), respectively. Whereas the concomitant formation of AMP is stimulated by the presence of dithiothreitol in a concentration dependent manner, the synthesis of p4A is only slightly inhibited by this compound. Previous treatment of the enzyme (E1) with iodoacetamide inhibited only partially the synthesis of p4A. p4A can substitute for ATP as substrate of the reaction to generate the ubiquityl adenylate complex. A small amount of diadenosine pentaphosphate (Ap5A) was also synthesized in the presence of p4A. PMID- 16256114 TI - In-hospital downgrading of the trauma team: Validation of the Academic Medical Center downgrading criteria. AB - BACKGROUND: To reduce overtriage of trauma patients while at the same time minimising undertriage, an in-hospital triage tool was developed with the purpose of reducing the initial full trauma team (downgrading) in a structured and evidence-based manner. This study evaluated the effect on overtriage rates by the AMC downgrading protocol (AMCDP) consisting of 24 criteria scored during the primary survey. PATIENTS AND METHODS: We prospectively investigated if any of the patients treated by the downgraded trauma team (DTT) were undertriaged by the protocol. All patients fulfilling the definition of severely injured (SI) patients but treated by the DTT were deemed undertriaged patients. Overtriage was measured by the percentage of patients treated by a full trauma team (FTT) while not classified as an SI patient. RESULTS: A total of 220 patients were eligible and triaged by the AMCDP. After triage, 95 patients (43%) were treated by the DTT while 125 patients (57%) were treated by the FTT. A total of 66 patients (30%) met one or more of the criteria for an SI patient. None of these patients were treated by the DTT. Of the 125 patients treated by the FTT, 59 patients were not defined as SI. CONCLUSION: For the entire study population no undertriage was found, while implementation of the AMCDP reduced overtriage in the entire study population from 70% to 26.8%. Similar trauma centres can benefit from implementing the AMC downgrading protocol. PMID- 16256115 TI - The use of regional anaesthesia in patients at risk of acute compartment syndrome. AB - A delay in the diagnosis of an acute compartment syndrome can be devastating to the patient. The increasing use of regional anaesthesia in the management of orthopaedic and trauma patients raises concerns about the potential for delay in the diagnosis of acute compartment syndrome. We undertook a postal survey to assess the usage of regional anaesthesia in patients with lower limb fractures. The study showed that regional anaesthesia is being used in patients at risk of compartment syndrome and without compartment pressure monitoring equipment being available. The anaesthetists questioned had seen cases of acute compartment syndrome being masked by regional anaesthesia. We recommend that there is an urgent need to establish joint guidelines between the orthopaedic and anaesthetic communities on the usage of regional anaesthesia in patients with lower limb fractures to reduce further morbidity from delays in the diagnosis of compartment syndrome. PMID- 16256117 TI - Mannose-binding lectin and vulvovaginal candidiasis. AB - OBJECTIVE: To investigate the effect of mannose-binding lectin (MBL) gene polymorphism on the immune system and the significance of vaginal MBL concentration in the pathogenesis of vulvovaginal candidiasis (VVC) and recurrent VVC (rVVC). PATIENTS AND METHODS: Mannose-binding lectin concentrations in CVL samples from 111 women were collected between August 2004 and November 2004, 51 with VVC, 6 with rVVC patients, and 54 healthy women. CVL MBL concentrations were determined by enzyme-linked immunosorbent assay, and MBL gene polymorphism was analyzed by polymerase chain reaction and the restriction fragment length polymorphism method. RESULTS: Cervicovaginal lavage MBL concentration (17.80 ng/mL) and gene mutation frequency (33.33%) were both significantly higher in women with VVC (P<0.01) than in controls (odds ratio [OR], 2.93; 95% confidence interval [CI], 1.14-7.57; P<0.05). On the other hand, MBL concentration was lower in women with rVVC (0.30 ng/mL) than in controls (1.28 ng/mL) (P<0.05), although MBL gene mutation frequency (83.33%) was significantly higher in women with rVVC than in controls (OR, 26.87; 95% CI, 2.76-261.65; P<0.01). CONCLUSIONS: The presence of VVC can increase vaginal MBL level, which may be an immune response against Candida albicans infection; in women with rVVC, the low level of MBL in the vagina caused by mutation in the MBL gene may play a role in the recurrence of the infection. PMID- 16256118 TI - The decision-making processes of nurses when extubating patients following cardiac surgery: an ethnographic study. AB - BACKGROUND: The movement towards research and evidence-based practice in health care demands that the best available evidence is applied to practice. At the same time, changes to role boundaries mean that nurses are assuming increased responsibility, especially in relation to decision making. While increasing, there has been limited consideration about the application of best evidence and decision making by nurses in the context of their clinical work. OBJECTIVES: This study sought to explore the realities of research and evidence-based practice through an examination of the decision making of nurses when extubating patients following cardiac surgery. DESIGN: The tradition of qualitative research and, more specifically, ethnography were used for the study. SETTING: Data were gathered over an 18-month period during 1998 and 1999 within a Cardiothoracic Intensive Care Unit (CICU). PARTICIPANTS: The sample comprised 43 nursing, 16 medical and two managerial staff. A purposive sample of five nurses, a cardiac surgeon, intensivist, CICU manager and Deputy Divisional Manager were included in interviews. METHODS: All staff were included in participant observation. Semi structured interviews were conducted with a purposive sample of nurses during the 6th and 14th months and with a purposive sample of other staff during the 16th month. Data were analysed using progressive focusing, data source triangulation and sensitising concepts to identify themes and categories. RESULTS: The findings indicated that, despite the use of an unwritten physiologically based protocol for weaning and extubation, factors other than best evidence were significant in nurses' decision making. A range of personal, cultural and contextual factors including relationships, hierarchy, power, leadership, education, experience and responsibility influenced their decision making. CONCLUSION: This study revealed, often disregarded, cultural, contextual and personal characteristics which combined to form a complex process of decision making. Providing new insight into research and evidence-based practice, the findings have implications for policy makers, educators, managers and clinicians and for the continued professional development of nursing. PMID- 16256119 TI - The good old days of nurse training: rose-tinted or jaundiced view? PMID- 16256122 TI - The beat-to-beat decay of cardiac contractility from highly potentiated levels is bi-exponential. AB - In order to determine the mode of beat-to-beat decay of contractility from very high levels, we studied the beat-by-beat decay of cardiac contractility following potentiation. Such decay curves are normally analysed using a mono-exponential decay function, which assumes that a fixed fraction of activator calcium ions is recirculated from one beat to the next. We postulated that there might be deviations from such a mono-exponential expression at high levels of contractility. In single sucrose-gap voltage clamp experiments of isolated ferret papillary muscle, we obtained very high contractility by potentiation due to prolonged depolarisations. We found a bi-exponential decay in 9 of 11 muscles studied, in which the initial decay is much faster than the subsequent slower decay, as judged by residual variance of least-squares exponential fitting and by analysis of covariance using a linear equation (force of beat versus force of previous beat), p = 0.0089. In the slower decay period (physiological range), the decay was identical to that following post-extrasystolic potentiation in the same muscles studied with conventional stimulation. PMID- 16256120 TI - Effects of bezafibrate on the expression of endothelial nitric oxide synthase gene and its mechanisms in cultured bovine endothelial cells. AB - OBJECTIVE: Peroxisome proliferator-activated receptors alpha (PPARalpha) is a target gene for atherosclerosis and cardiovascular diseases. However, effects of PPARalpha on endothelial nitric oxide synthase (eNOS) remain unknown. We investigated the eNOS regulation by bezafibrate, a ligand of PPARalpha, and involved signaling pathways. METHODS AND RESULTS: Firstly, in cultured bovine aorta endothelial cells (BAEC), bezafibrate significantly upregulated eNOS at protein, mRNA levels and NO production, respectively, in a concentration dependent fashion (50-200muM). Next, the effects of bezafibrate on signal pathways and eNOS mRNA stability in BAEC were investigated. Results showed that bezafibrate induced phosphorylation of MAPK. Inhibitors of PPARalpha, PI3 kinase and MAPK, respectively, markedly attenuate bezafibrate-induced upregulation of eNOS. Bezafibrate incubation increased eNOS mRNA half-life, activated eNOS promoter, enhanced phosphorylation of eNOS ser-1179 site, and decreased phosphorylation of eNOS thr-497 site via activating ERK and Akt. CONCLUSIONS: Bezafibrate can upregulate eNOS expression, enhance phosphorylation of eNOS ser 1179, increase NO production and transcription level and stability of eNOS mRNA through pathway dependent of PPARalpha and nongenomic effects mediated by MAPK and PI3K pathways. Hence, PPARalpha ligands exert direct benefits on vessel endothelial functions through an increase in eNOS expression level and phosphorylation of eNOS ser-1179. This mechanism provides additional anti atherosclerotic and anti-hypertension benefits of bezafibrate in addition of lipid-lowering effects. PMID- 16256121 TI - Flexion-relaxation response to gravity. AB - The objective of this report was to study the influence of the orientation of gravitational loading on the behavior of anterior and posterior trunk muscles during anterior trunk flexion-extension. Participants (N=13) performed five (5) cycles of trunk flexion-extension while standing with gravity parallel to the body axis and five (5) cycles while in the supine condition (e.g. sit-ups) with gravity perpendicular to the body axis. Surface electromyographic (EMG) patterns from lumbar paraspinal, rectus abdominis, external oblique, rectus femoris, semimembranosis, and biceps femoris muscles were analyzed during each condition. EMG signals were synchronized with lumbar flexion and trunk inclination angles. Flexion-extension from the standing position resulted in a myoelectric silent period of the lumbar posterior muscles (e.g. flexion-relaxation phenomena (FRP)) as well as the hamstring muscles through deep angles during which activity was observed in abdominal muscles. Flexion-extension during sit-ups, however, resulted in a myoelectric silent period of the abdominal muscles and the quadriceps through deep angles during which the lumbar posterior muscles were active. In this condition, the FRP was not observed in posterior muscles. The new findings demonstrate the profound impact of the orientation of the gravity vector on the FRP, the abdominal muscles reaction to gravitational loads during sit-ups and its relationships with lumbar antagonists and thigh musculature. The new findings suggest that gravitational moments requirements dominate the FRP through the prevailing kinematics, load sharing and reflex activation-inhibition of muscles in various conditions. Lumbar kinematics or fixed sensory motor programs by themselves, however, are not the major contributor to the FRP. The new findings improve our insights into spinal biomechanics as well as understanding and evaluating low back disorders. PMID- 16256123 TI - Isolated fibrillar damage in tendons stimulates local collagenase mRNA expression and protein synthesis. AB - The etiology of repetitive stress injuries in tendons has not been clearly identified. While minor trauma has been implicated as an inciting factor, the precise magnitude and structural level of tissue injury that initiates this degenerative cascade has not been determined. The purpose of this study was to determine if isolated tendon fibril damage could initiate an upregulation of interstitial collagenase (MMP13) mRNA and protein in tendon cells associated with the injured fibril(s). Rat tail tendon fascicles were subjected to in vitro tensile loading until isolated fibrillar damage was documented. Once fibrillar damage occurred, the tendons were immediately unloaded to 100g and maintained at that displacement for 24h under tissue culture conditions. In addition, non injured tendon fascicles were maintained under unloaded (stress-deprived) conditions in culture for 24h to act as positive controls. In situ hybridization or immunohistochemistry was then performed to localize collagenase mRNA expression or protein synthesis, respectively. Fibrillar damage occurred at a similar stress (41.13+/-5.94MPa) and strain (13.24+/-1.94%) in the experimental tendons. In situ hybridization and immunohistochemistry demonstrated an upregulation of interstitial collagenase mRNA and protein, respectively, in only those cells associated with the damaged fibril(s). In the control (stress deprived) specimens, collagenase mRNA expression and protein synthesis were observed throughout the fascicle. The results suggest that isolated fibrillar damage and the resultant upregulation of collagenase mRNA and protein in this damaged area occurs through a mechanobiological understimulation of tendon cells. This collagenase production may weaken the tendon and put more of the extracellular matrix at risk for further damage during subsequent loading. PMID- 16256124 TI - Stress analysis of carotid plaque rupture based on in vivo high resolution MRI. AB - Atheromatous carotid plaque rupture is responsible for the majority of ischaemic strokes in the developed world. Plaque rupture has been associated with plaque morphology, plaque components' properties, inflammation and local stress concentration. High resolution multi-spectral magnetic resonance imaging (MRI) has allowed the plaque components to be visualized in vivo. This study combined the recent advances in finite element analysis (FEA) and MRI, and performed stress analysis of five vulnerable carotid plaques based on the geometry derived from in vivo MRI. Image segmentation was based on multi-spectral MRI and co registered with histology for plaque characterization. Plaque fibrous cap, lipid pool and vessel wall were modelled as isotropic, incompressible hyperelastic materials undergoing large deformation under pulse pressure loading. High stress concentrations were predicted at the shoulders and the thinnest fibrous cap regions of the plaque, and the mean maximal stresses were found to be higher in the ruptured plaques (683.3 kPa) than those in the unruptured plaques (226.9 kPa). The effect of the relative stiffness of fibrous cap to lipid pool on the stress within the cap itself was studied. It was shown that larger relative stiffness of fibrous cap to lipid pool resulted in higher stress within the cap. Thus, it is likely that high stress concentrations in vulnerable plaque may cause plaque rupture and lead to acute ischaemic sequelae. A combination of in vivo high resolution MRI and FEA could potentially act as a useful tool to assess plaque vulnerability and risk stratify patients with carotid atheroma. PMID- 16256125 TI - Cyclic strain increases fibroblast proliferation, matrix accumulation, and elastic modulus of fibroblast-seeded polyurethane constructs. AB - Rapid induction of matrix production and mechanical strengthening is essential to the development of bio-artificial constructs for repair and replacement of load bearing connective tissues. Toward this end, we describe the development of a mechanical bioreactor and its application to investigate the influence of cyclic strain on fibroblast proliferation, matrix accumulation, and the mechanical properties of fibroblast-seeded polyurethane constructs (FSPC). Human fibroblasts were cultured in 10% serum-containing conditions within three-dimensional, porous elastomeric substrates under static conditions and a model regime of cyclic strain (10% strain, 0.25 Hz, 8 h/day), with and without ascorbic acid supplementation. After one week, the combination of cyclic strain and ascorbic acid resulted in significantly increased construct elastic modulus (>110%) relative to either condition alone. In contrast, cyclic strain alone was sufficient to stimulate significant increases in fibroblast proliferation. Mechanical strengthening of FSPCs was accompanied by increased type I collagen and fibronectin matrix accumulation and distribution, and significantly increased gene expression for type I collagen, TGFbeta-1, and CTGF. These results suggest that strain-induced conditioning in vitro leads to mechanical strengthening of fibroblast/material constructs, most likely resulting from increased collagen matrix deposition, secondary to strain-induced increases in cytokine production. PMID- 16256126 TI - On the implications of interpreting the stability index: a spine example. PMID- 16256127 TI - Determination of sixteen polycyclic aromatic hydrocarbons in aqueous and solid samples from an Italian wastewater treatment plant. AB - A robust procedure for the determination of 16 US EPA PAHs in both aqueous (e.g. wastewaters, industrial discharges, treated effluents) and solid samples (e.g. suspended solids and sludge) from a wastewater treatment plant (WWTP) is presented. Recovery experiments using different percentages of organic modifier, sorbents and eluting solvent mixtures were carried out in Milli-Q water (1000 mL) spiked with a mixture of the PAH analytes (100 ng/L of each analyte). The solid phase extraction (SPE) procedures applied to spiked waste water samples (1000 mL; 100 ng/L spiking level) permitted simultaneous recovery of all the 16PAHs with yields >70% (6-13% RSD). SPE clean up procedures applied to sewage and stabilized sludge extracts, showed percent recoveries in the range 73-92% (7-13% RSD) and 71 89% (7-12% RSD), respectively. The methods were used for the determination of PAHs in aqueous and solid samples from the WWTP of Fusina (Venice, Italy). Mean concentrations, as the sum of the 16PAHs in aqueous and suspended solid samples, were found to be approx. in the 1.12-4.62 microg/L range. Sewage and stabilized sludge samples contained mean PAH concentrations, as sum of 16 compounds, in the concentration range of 1.44-1.26 mg/kg, respectively. Extraction and clean up procedures for sludge samples were validated using EPA certified reference material IRM-104 (CRM No. 912). Instrumental analyses were performed by coupling HPLC with UV-diode array detection (UV-DAD) and fluorescence detection (FLD). PMID- 16256128 TI - Influence of the protein oligomericity on final yield after affinity tag removal in purification of recombinant proteins. AB - The new aspect concerning the applicability of histidine and other affinity tags for the purification of oligomeric proteins, with particular emphasis on cleavage efficiency and final yield, is presented in this study. The final yield depends on both the cleavage efficiency and the degree of oligomerization of the protein. Cleavage procedures that are good enough for monomeric proteins can be problematic for oligomeric proteins. Random distribution of uncleaved or partially cleaved affinity tags among oligomers is the main cause of reduced yields. A trimeric protein, tumour necrosis factor alpha (TNF-alpha), bearing different histidine tags, was used as a model protein to explore and confirm this theoretical concept. Analysis of mixed TNF trimers, prepared from tag-free TNF doped with various amounts of histidine-tagged TNF, revealed an increased retention of the trimeric protein on immobilized metal-ion affinity chromatography (IMAC) columns. When 20% of histidine-tagged TNF was added, more than 50% of the protein was retained on the IMAC column. Thus, the applicability of histidine- and other affinity tags for purifying oligomeric proteins is significantly prejudiced in the case of higher oligomers. Various histidine-tags were fused to the N-terminus of full-length TNF-alpha and to the truncated form (dN6) of TNF-alpha. Two-step IMAC separation was used for purification. In the first step, IMAC-1, over 95% purity of histidine-tagged protein was achieved in all cases. Endo- and exoproteolytic removal of histidine tags with enterokinase (EKmax) and aminodipeptidase (DAPase) was studied and the major parameters affecting cleavage efficiency, microheterogeneity and final yield are critically discussed. IMAC-2 was used as the second and final step for removing the cleavage enzyme, cleaved tags, unprocessed protein and some other impurities. Selection of the optimal cleavage enzyme depends on the amino acid composition of the N terminus and the intended use of the purified protein. The main conclusion is that special caution should be taken when introducing affinity tags to oligomeric proteins, with the final goal to produce pure, tag-free protein with acceptable yields. Given the same enzyme cleavage efficiency one can expect progressively reduced final protein yields with increasing degree of oligomerization. This should be considered as a general rule. PMID- 16256129 TI - Acid dye adsorption onto cationized polyamide fibres. Modeling and consequent interpretations of model parameter behaviours. AB - Experimental adsorption isotherms of four acid dyes named Acid Blue 25, Acid Yellow 99, Reactive Yellow 23, and Acid Blue 74 from aqueous solution onto cationized nylon-6,6 have been analyzed using a double layer adsorption model. The parameters involved in the analytical expression of this model such as the number or fraction of adsorbed dye molecule per site, n, the number of receptor sites per gram of adsorbent, N(M), and the concentration at half-saturation, c1/2, are determined from adsorption isotherms at four temperatures between 293 and 353 K. The evolution of these parameters with temperature is discussed in relation with adsorption process and the behaviours of the different dyes taking into account their particular structure. The results are compared with those already published dealing with the adsorption of these same dyes onto cationized cotton. The configurational entropy at various temperatures has been studied. This parameter allowed to deduce some results related to the evolution of the disorder during the adsorption process. PMID- 16256130 TI - Mixed self-assembled monolayers (SAMs) consisting of methoxy-tri(ethylene glycol) terminated and alkyl-terminated dimethylchlorosilanes control the non-specific adsorption of proteins at oxidic surfaces. AB - Monolayers from the newly synthesized compound methoxy-tri(ethylene glycol) undecenyldimethylchlorosilane (CH3O(CH2CH2O)3(CH2)11Si(CH3)2Cl, MeO(EG)3C11DMS) and dodecyldimethylchlorosilane (DDMS), both pure and mixed, were prepared by self-assembly from organic solution in the presence of an organic base. The films obtained were characterized by advancing and receding contact angle measurements and ellipsometry to confirm the formation of self-assembled monolayers (SAMs). The resulting data on the covalently attached dimethylsilanes were compared to known oligo(ethylene glycol) (OEG)-terminated SAM systems based on terminal alkenes, thiolates or trihydrolyzable silanes. The composition of the mixed SAMs was found to depend directly and linearly on the composition of the silanization solution. Enhanced protein repellent properties were found for the SAMs using a variety of proteins, including the Ras Binding Domain (RBD), a protein with high relevance for cancer diagnostics. Roughly a RBD protein monolayer amount was adsorbed to silicon oxide surfaces silanized with DDMS or non-silanized silicon wafers, and in contrast, no RBD was adsorbed to surfaces silanized with MeO(EG)3C11DMS or to mixed monolayers consisting of DDMS and MeO(EG)3C11DMS if the content of OEG-silane overcame a critical content of X(EG) approximately 0.9. PMID- 16256131 TI - Effects of the number of fatty acid residues on the phase behaviors of decaglycerol fatty acid esters. AB - The effects of the number of fatty acid residues (n) in decaglycerol fatty acid esters, i.e., decaglycerol laurates (abbreviated to (C11)nG10), on the phase behaviors of three laurate esters, (C11)1.9G10, (C11)2.7G10, and (C11)3.4G10, were investigated. The unreacted decaglycerol remaining in each ester was removed by liquid extraction before use. (C11)1.9G10 formed hexagonal liquid crystals in aqueous solutions, while (C11)2.7G10 and (C11)3.4G10, which are more hydrophobic than (C11)1.9G10, formed lamellar liquid crystals. The cloud point in aqueous solution was measured for mixtures of these three esters. The cloud phenomenon was observed when the weight ratio of hydrophilic groups to the total surfactant (WH/WS) was around 0.6. The cloud point shifted to a markedly higher temperature, even with a slight increase in the WH/WS ratio. The solubilization abilities of (C11)nG10 for the oils m-xylene and (R)-(+)-limonene were also examined. When the WH/WS ratio was between 0.60 and 0.64, (C11)nG10 formed microemulsions and lyotropic liquid crystals in the presence of water and the oils. These self organized structures were stable, even above 90 degrees C. It is concluded that the phase behavior of (C11)nG10 are insensitive to temperature, but strongly dependent on both the WH/WS ratio and the number of fatty acid residues (n). PMID- 16256133 TI - Flow cytometric determination of mitochondrial membrane potential changes during apoptosis of T lymphocytic and pancreatic beta cell lines: comparison of tetramethylrhodamineethylester (TMRE), chloromethyl-X-rosamine (H2-CMX-Ros) and MitoTracker Red 580 (MTR580). AB - The mitochondria-specific dyes, TMRE, H2-CMX-Ros and MTR580 were determined for their suitability to measure mitochondrial potential changes of the T cell leukemia cell line Jurkat and insulin-secreting beta cell line NIT-1 during apoptosis. Both freshly harvested Jurkat and NIT-1 cells induced to undergo apoptosis displayed poor retention of the potential-sensitive, intrinsically fluorescent dye, TMRE. Treatment with formaldehyde or paraformaldehyde completely abolished TMRE uptake in both cell types regardless of apoptosis induction. Interestingly, freshly harvested apoptotic Jurkat cells exhibited lower retention of H2-CMX-Ros, indicating marked reduction in the oxidative status of lymphoid cells during apoptosis. This is in contrast to NIT-1 cells which failed to display significant reduction in H2-CMX-Ros retention after anoikis induction. Paraformaldehyde treatment reduced the retention of H2-CMX-Ros in live Jurkat cells but still allowed the discrimination of apoptotic cells which poorly retained H2-CMX-Ros. However, live Jurkat cells lost their ability to retain H2 CMX-Ros after formaldehyde treatment. In contrast, treatment with paraformaldehyde or formaldehyde did not have significant impact on the retention of H2-CMX-Ros in both live and apoptotic NIT-1 cells. The uptake of MTR580 was independent of mitochondrial membrane potential in both T and beta cell lines. However, MTR580 was comparable to H2-CMX-Ros for confocal microscopic analysis of apoptotic Jurkat cells following fixation with formaldehyde and cell permeabilization. These data demonstrate that while TMRE and H2-CMX-Ros are suitable for determining mitochondrial membrane potential changes during apoptosis in lymphoid cells, only TMRE is suitable for such analysis in beta cells. Both H2-CMX-Ros and MTR580 proved to be suitable for confocal imaging of mitochondria. PMID- 16256134 TI - Activity modulation in cockroach sensillum: the role of octopamine. AB - The plasticity of sensory perception is provided partially by modulation of receptor cells. The electrical activity of American cockroach chemoreceptor cells in response to sex pheromone was measured under the influence of octopamine treatment and tracheal anoxia. Both experimental procedures caused decreased electroantennograms but affected spike activity differently: octopamine treatment increased firing rate, whereas anoxia decreased it. Spike frequency under octopamine treatment was elevated in response to pheromone stimulation and at background activity. Experiments with perfusion of isolated antennae showed a direct effect of octopamine on spike activity of pheromone sensilla, and excluded the possibility of indirect effects via octopamine-dependent release of other biologically active substances. The suggested mechanism of octopamine action is receptor cell membrane depolarization. PMID- 16256135 TI - Genome comparison of Pseudomonas aeruginosa large phages. AB - Pseudomonas aeruginosa phage EL is a dsDNA phage related to the giant phiKZ-like Myoviridae. The EL genome sequence comprises 211,215 bp and has 201 predicted open reading frames (ORFs). The EL genome does not share DNA sequence homology with other viruses and micro-organisms sequenced to date. However, one-third of the predicted EL gene products (gps) shares similarity (Blast alignments of 17 55% amino acid identity) with phiKZ proteins. Comparative EL and phiKZ genomics reveals that these giant phages are an example of substantially diverged genetic mosaics. Based on the position of similar EL and phiKZ predicted gene products, five genome regions can be delineated in EL, four of which are relatively conserved between EL and phiKZ. Region IV, a 17.7 kb genome region with 28 predicted ORFs, is unique to EL. Fourteen EL ORFs have been assigned a putative function based on protein similarity. Assigned proteins are involved in DNA replication and nucleotide metabolism (NAD+-dependent DNA ligase, ribonuclease HI, helicase, thymidylate kinase), host lysis and particle structure. EL-gp146 is the first chaperonin GroEL sequence identified in a viral genome. Besides a putative transposase, EL harbours predicted mobile endonucleases related to H-N-H and LAGLIDADG homing endonucleases associated with group I intron and intein intervening sequences. PMID- 16256136 TI - Lactated Ringer's solution and hypertonic saline improve survival in uncontrolled hemorrhagic shock in female rats in metestrus. AB - BACKGROUND: In the present investigation the effect of fluid treatment in uncontrolled hemorrhagic shock after massive splenic injury (MSI) was comparatively studied in male and female rats. MATERIALS AND METHODS: The anesthetized animals were randomly divided into three groups: in group 1 MSI was induced in males, in group 2 MSI was induced in females in proestrus, in group 3 MSI was induced in females in metestrus. Each group was divided into four subgroups: a) Sham-operated, b) MSI untreated (UT), c) MSI treated with 40 ml/kg lactated Ringer's solution (RL), and d) MSI treated with 5 ml/kg NaCl 7.5% (HTS). RESULTS: Total blood loss (TBL) in groups 1b, 2b, and 3b was 31.7 +/- 3.6%, 33.1 +/- 2.6%, and 36.7 +/- 2.6%, respectively, and mean survival time (MST) was 143.7 +/- 25.3 min, 174.8 +/- 10.4 min, and 67.8 +/- 11.4 min (P < 0.01 versus group 2b), respectively. TBL in groups 1c, 2c, and 3c increased to 52.4 +/- 5.5% (P < 0.02 versus UT), 48.6 +/- 1.6% (P < 0.02 versus UT), and 48.8 +/- 4.1% (P < 0.02 versus UT), respectively, and MST decreased to 126 +/- 19.4 min, (P < 0.05 versus UT), and 136.8 +/- 13.0 min (P < 0.05 versus UT) in groups 1c and 2c, respectively, and increased in group 3c to 120.4 +/- 23.3 min (P < 0.05 versus UT). TBL in groups 1d, 2d, and 3d was 31.3 +/- 4.8%, 38.0 +/- 4.2%, and 40.6 +/- 3.7%, respectively, and MST increased to 198.5 +/- 13.9 min (P < 0.05 versus UT) in group 1d, decreased to 128.4 +/- 17.2 min (P < 0.01) in group 2d, and increased to 102.6 +/- 19.0 min (P < 0.002 versus group 1d) in group 3d. CONCLUSIONS: RL infusion significantly increased blood loss in all three groups, reduced survival time in males and female rats in proestrus, but significantly improved survival in females in metestrus. HTS treatment did not alter blood loss in all three groups, but significantly improved survival in females in metestrus and males. PMID- 16256137 TI - Identification of differentially expressed genes in papillary thyroid carcinomas with and without rearrangements of the tyrosine kinase receptors RET and/or NTRK1. AB - BACKGROUND: The transforming capacities of RET and/or NTRK1 chimeric oncogenes as well as the molecular background of non-rearranged papillary thyroid carcinomas (PTCs) remain to be elucidated. To assess altered gene expression, we examined PTCs with and without tyrosine kinase receptor rearrangements by mRNA differential display (DD). MATERIALS AND METHODS: Six of 13 PTCs examined harbored RET chimeras (3x RET/PTC1, 1x RET/PTC3) and/or NTRK1 chimeras (2x trk, 1x TRK-T3, 2 unknown TRK hybrids). The method of DD analysis was refined by a novel fragment-recovery technique using a high-performance fluorescence scanner. RESULTS: Of 500 up- or down-regulated mRNA transcripts, 19 selected fragments were recovered, cloned, sequenced, and identified. The accuracy and high degree of reproducibility of the method was demonstrated. Differential expression of gene products with potential association to cell proliferation or tumor progression was observed, such as 14-3-3beta and Rab27a. Moreover, several gene products with unknown functions were demonstrated in PTCs bearing RET or NTRK1 hybrids versus rearrangement-negative PTCs, including a homologue of the Ig kappa light chain constant region. CONCLUSIONS: Candidate transcripts with presumed tumorigenic potential in other solid tumors may prove to be relevant in the progression of PTCs, too. Most promising is the isolation of several differentially expressed, yet unknown, genes that may open new insights in the pathogenesis or progression of PTC. PMID- 16256138 TI - Angiotensin II subtype AT1 and AT2 receptors regulate microvascular hydraulic permeability via cAMP and cGMP. AB - INTRODUCTION: Angiotensin II receptor subtypes (AT1 and AT2) have been shown to modulate microvascular fluid leak. However, their intracellular signal transduction pathways have not been elucidated. We hypothesized that AT1 activation exerts its permeability-increasing effect by provoking cGMP synthesis and inducing cAMP degradation and that AT2 activation decreases fluid leak by stimulating cAMP synthesis and enhancing cGMP degradation. METHODS: Using a microcannulation technique, hydraulic permeability (Lp) was measured in rat mesenteric venules. The messenger signal transduction of ATI was studied during continuous perfusion with the AT1 agonist, Sar1 plus either 1) a cGMP synthesis inhibitor, LY83583, or 2) an inhibitor of cAMP degradation, Rolipram. Likewise, AT2 signal transduction was studied with the AT2 agonist, CGP42112A, plus either 1) a cAMP synthesis inhibitor, dideoxyadenosine, or 2) an inhibitor of cGMP degradation, Zaprinast. Lp values are represented as mean +/- SEM x 10(-7) cm/s/cm H2O. For each group n = 6. RESULTS: Inhibition of cGMP synthesis blunted the permeability-increasing effect of AT1 agonism and decreased the peak Lp from 4.91 +/- 0.25 to 2.30 +/- 0.10 (P < 0.001). Inhibition of cAMP degradation also reduced the effect of AT1 agonism on peak L(p) from 2.25 +/- 0.22 to 1.30 +/- 0.13 (P < 0.001). Meanwhile, cAMP synthesis inhibition completely blocked the permeability-decreasing effect of AT2 agonism during which Lp increased from a baseline of 0.92 +/- 0.08 to a peak of 4.38 +/- 0.20 (P < 0.001). During inhibition of cGMP degradation, AT2 activation was able to decrease peak Lp from 2.26 +/- 0.15 to 1.46 +/- 0.05 (P < 0.001). CONCLUSIONS: When cGMP synthesis and cAMP degradation were inhibited, the effect on fluid leak by AT1 activation was blunted. Inhibition of cAMP synthesis completely blocked the effect of AT2 activation on fluid leak, while AT2 activation continued to decrease fluid leak despite inhibition of cGMP degradation. The AT1 receptor appears to increase fluid leak by stimulating both cGMP synthesis and cAMP degradation, while the AT2 receptor decreases fluid leak by stimulating cAMP synthesis, but not cGMP degradation. PMID- 16256139 TI - Effect of ischemic preconditioning on rat liver microcirculation monitored with laser Doppler flowmetry. AB - BACKGROUND: Ischemic preconditioning (IP) may protect the liver from ischemia reperfusion (I-R) injury during liver resection. This study investigated the effect of IP on hepatic microcirculation (HM) and analyzed the objective parameters of the HM using laser Doppler flowmetry (LDF). METHODS: We used male Wistar rats (250-280 g) that underwent normothermic, segmental liver ischemia. The animals were divided into eight groups: 30, 45, 60, and 90 min of ischemia with, or without, IP. Five minutes ischemia and 10 min reperfusion, in two cycles, were used to elicit IP. Changes of the hepatic microcirculation were studied by LDF with on-line computer monitoring and processing. Histological alterations, liver enzymes, bilirubin, and TNF-alpha level were all measured simultaneously. RESULTS: Reperfusion was assessed by post-ischemia flux plateau maximum (PM) and by the area under the reperfusion-curve (RA). Both PM and RA are inversely correlated with the duration of ischemia. The groups with IP had significantly (P < 0.05) higher flow values than groups without preconditioning. IP before liver ischemia resulted in significantly (P < 0.05) lower TNF-alpha levels at the end of the 30-min reperfusion. Lower serum ALT, LDH, and bilirubin levels could only be observed at 45 and 60 min I-R during the first post operative day. On the seventh post-operative day there were no significant differences between the I-R and IP + IR groups in any of these parameters. CONCLUSION: The benefit of ischemic preconditioning on hepatic microcirculation was well demonstrated with this method, which has never been described before, in this context. Changes in hepatic microcirculation can be precisely investigated by laser Doppler flowmetry using the standardization and transformation described in this paper. PMID- 16256141 TI - Incidental findings in magnetic resonance imaging of the brains of healthy young men. AB - BACKGROUND AND PURPOSE: To determine the frequency of serious intracranial abnormalities in a healthy young male population. METHODS: Cranial MRI of 2,536 healthy young males, mean age 20.5 years, all applicants for military flying duties in the German Air Force. RESULTS: The authors report a variety of morphological abnormalities in the brains of a large population of healthy young males, providing data on disease prevalence. Arachnoid cysts were found in 1.7% (95% CI 1.2 to 2.3%), vascular abnormalities in 0.51% (95% CI 0.29 to 0.9%), and intracranial tumors in 0.47% (95% CI 0.26 to 0.85%) of the applicants. No cerebral aneurysms were found. CONCLUSION: The prevalence of primary brain tumors seems to be higher, whereas the prevalence of intracranial aneurysms is lower than expected. Only a small percentage of the detected abnormalities require urgent medical attention. PMID- 16256140 TI - Platelet amyloid precursor protein processing: a bio-marker for Alzheimer's disease. AB - The amyloid precursor protein (APP) in brain is processed either by an amyloidogenic pathway by beta-secretase and gamma-secretase to yield Abeta (beta amyloid 4 kDa) peptide or by alpha-secretase within the beta-amyloid domain to yield non-amyloidogenic products. We have studied blood platelet levels of a 22 kDa fragment containing the Abeta (beta-amyloid 4 kDa) peptide, beta-secretase (BACE1), alpha-secretase (ADAM10), and APP isoform ratios of the 120-130 kDa to 110 kDa peptides from 31 Alzheimer's disease (AD) patients and 10 age-matched healthy control subjects. We found increased levels of Abeta4, increased activation of beta-secretase (BACE1), decreased activation of alpha-secretase (ADAM10) and decreased APP ratios in AD patients compared to normal control subjects. These observations indicate that the blood platelet APP is processed by the same amyloidogenic and non-amyloidogenic pathways as utilized in brain and that APP processing in AD patients is altered compared to control subjects and may be a useful bio-marker for the diagnosis of AD, the progression of disease and for monitoring drug responses in clinical trials. PMID- 16256142 TI - Searching on patch networks using correlated random walks: space usage and optimal foraging predictions using Markov chain models. AB - We describe a novel representation of a discrete correlated random walk as the transition matrix of a Markov chain with the displacements as the states. Such a representation makes it possible to utilize results from the theory of absorbing Markov chains, to make biologically interesting predictions without having to resort to Monte Carlo simulations. Our motivation for constructing such a representation is to explore the relationship between the movement strategy of an animal searching for resources upon a network of patches, and its consequent utilization of space and foraging success. As an illustrative case study, we have determined the optimal movement strategy and the consequent usage of space for a central place forager utilizing a continuous movement space which is discretized as a hexagonal lattice. The optimal movement strategy determines the size of the optimal home range. In this example, the animal uses mnemokinesis, which is a sinuosity regulating mechanism, to return it to the central place. The movement strategy thus refers to the choice of the intrinsic path sinuosity and the strength of the mnemokinetic mechanism. Although the movement space has been discretized as a regular lattice in this example, the method can be readily applied to naturally compartmentalized movement spaces, such as forest canopy networks. This paper is thus an attempt at incorporating results from the theory of random walk-based animal movements into Foraging Theory. PMID- 16256143 TI - Multifractality in intracellular enzymatic reactions. AB - Enzymatic kinetics adjust well to the Michaelis-Menten paradigm in homogeneous media with dilute, perfectly mixed reactants. These conditions are quite different from the highly structured cell plasm, so applications of the classic kinetics theory to this environment are rather limited. Cytoplasmic structure produces molecular crowding and anomalous diffusion of substances, modifying the mass action kinetic laws. The reaction coefficients are no longer constant but time-variant, as stated in the fractal kinetics theory. Fractal kinetics assumes that enzymatic reactions on such heterogeneous media occur within a non-Euclidian space characterized by a certain fractal dimension, this fractal dimension gives the dependence on time of the kinetic coefficients. In this work, stochastic simulations of enzymatic reactions under molecular crowding have been completed, and kinetic coefficients for the reactions, including the Michaelis-Menten parameter KM, were calculated. The simulations results led us to confirm the time dependence of michaelian kinetic parameter for the enzymatic catalysis. Besides, other chaos related phenomena were pointed out from the obtained KM time series, such as the emergence of strange attractors and multifractality. PMID- 16256145 TI - Organic matter and nutrient inputs to the Humber Estuary, England. AB - Estuaries are sinks for organic matter and nutrients entering both from their catchments and also from the adjacent lands and urban areas and in turn they are sources of such materials to the adjacent coast. The present paper quantifies the relative amounts of natural and anthropogenic organic matter and nutrients entering the Humber Estuary, Eastern England, including the allochthonous and autochthonous materials, those from urban and industrial sewage and from the catchment drainage of arable land. These data thus give a budget for the estuary which in turn answers questions fundamental to the management of the estuary. The estimations within the study have been carried out against a background of designating estuaries under the European Union Urban Waste-water Treatment Directive and the EU Nitrates Directive. The assessment has particularly addressed the question, related to the former Directive, of whether the Humber Estuary is eutrophic or likely to become eutrophic unless management measures are taken. Thus the paper indicates the nature and value of control measures such as treatment plant upgrading and the designation of Nitrate Vulnerable Zones. The paper includes the recent national and European discussions on the designation of areas under these Directives. Finally, the study has allowed a quantification of the present organic inputs to the estuary in comparison to those entering prior to large scale land-claim which had removed natural organic-producing wetlands. PMID- 16256144 TI - Profiling differential gene expression of corals along a transect of waters adjacent to the Bermuda municipal dump. AB - A coral cDNA array containing 32 genes was used to examine the gene expression profiles of coral populations located at four sites that varied with distance from a semi-submerged municipal dump in Castle Harbour, Bermuda (previously identified as a point source of anthropogenic stressors). Genes on the array represent transcripts induced under controlled laboratory conditions to a variety of stressors both natural (temperature, sediment, salinity, darkness) and xenobiotic (heavy metals, pesticides, PAH) in origin. The gene expression profiles produced revealed information about the types of stressors. Consistent with other studies undertaken in Castle Harbour, the coral cDNA array detected responses to heavy metals, sedimentation, as well as oxidative stress. PMID- 16256146 TI - Influence of the oceanographic conditions during spring 2003 on the transport of the Prestige tanker fuel oil to the Galician coast. AB - Hydrographic data collected during the cruise HIDROPRESTIGE were combined with meteorological and dynamic data provided by remote sensors and drifting/moored buoys, to describe the surface circulation of the Northern Iberian basin in March April 2003. Sea surface winds transported the floating Prestige oil slicks from the sinking area to the continental slope off the Rias Baixas in 1/2 month: the surface current intensity was 2% of the wind intensity and it was rotated clockwise 5 degrees from the wind direction. Mesoscale cyclonic and anticyclonic structures west of 10 degrees W increased the residence time of oil patches in the Northern Iberian basin, as compared with the expected southwards flow of the Iberian current (IC). On the other hand, the Iberian poleward current (IPC) formed a marked surface front with coastal waters, preventing the entry of fuel oil into the rias. PAHs in the surface layer during the cruise were <0.5 microgL( 1), except in the Galicia bank (approximately 1 microg L(-1); where the Prestige tanker was still leaking) and the vicinity of Cape Fisterra (approximately 1.5 microg L(-1); where the convergence front between the IPC and coastal waters vanished). PMID- 16256148 TI - Time dependent calibration of a sediment extraction scheme. AB - Sediment extraction methods to quantify metal concentration in aquatic sediments usually present limitations in accuracy and reproducibility because metal concentration in the supernatant is controlled to a large extent by the physico chemical properties of the sediment that result in a complex interplay between the solid and the solution phase. It is suggested here that standardization of sediment extraction methods using pure mineral phases or reference material is futile and instead the extraction processes should be calibrated using site specific sediments before their application. For calibration, time dependent release of metals should be observed for each leachate to ascertain the appropriate time for a given extraction step. Although such an approach is tedious and time consuming, using iron extraction as an example, it is shown here that apart from quantitative data such an approach provides additional information on factors that play an intricate role in metal dynamics in the environment. Single step ascorbate, HCl, oxalate and dithionite extractions were used for targeting specific iron phases from saltmarsh sediments and their response was observed over time in order to calibrate the extraction times for each extractant later to be used in a sequential extraction. For surficial sediments, an extraction time of 24 h, 1 h, 2 h and 3 h was ascertained for ascorbate, HCl, oxalate and dithionite extractions, respectively. Fluctuations in iron concentration in the supernatant over time were ubiquitous. The adsorption desorption behavior is possibly controlled by the sediment organic matter, formation or consumption of active exchange sites during extraction and the crystallinity of iron mineral phase present in the sediments. PMID- 16256147 TI - Ecological consequences of dredged material disposal in the marine environment: a holistic assessment of activities around the England and Wales coastline. AB - This study provides a holistic perspective on the ecological effects of dredged material disposal, both intertidally and subtidally. A number of numerical techniques (univariate, distributional, multivariate and meta-analysis) were used to assess impacts at 18 different disposal sites. The analyses revealed that ecological effects associated with dredged material disposal were dependent on the numerical techniques used, and that impacts were disposal-site specific. Disposal-site communities were generally faunistically impoverished to varying degrees, and impacts following intertidal placement were comparable to those of subtidal placement. We conclude that any assessment of the consequences of dredged material disposal to the coastal environment must take account of site specific variation in prevailing hydrographic regimes and in ecological status, along with information on the disposal activity itself (mode, timing, quantity, frequency and type of material). As would be expected, variability in the latter presents a significant challenge in attempts to generalise about environmental and ecological impacts. PMID- 16256149 TI - Two novel mutations of the NOTCH3 gene in Korean patients with CADASIL. AB - Mutations in the NOTCH3 gene (NOTCH3) are responsible for cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL), an adult-onset hereditary angiopathy leading to ischemic episodes, vascular dementia and other neurologic deficits. All mutations of NOTCH3 described so far are strictly stereotyped, leading to the gain or loss of a cysteine residue in a given epidermal growth factor (EGF)-like repeat of NOTCH3. We report two novel mutations of NOTCH3, R587C and C988Y, each resulting in an odd number of cysteine residues in an EGF-like repeat of NOTCH3. We identified these mutations in two unrelated Korean families with CADASIL, who presented with magnetic resonance imaging (MRI) abnormalities typical of CADASIL. These findings confirm that mutations in NOTCH3 are associated with the pathogenesis of CADASIL across different ethnic backgrounds. PMID- 16256151 TI - Rotation or translation of auditory space in neglect? A case study of chronic right-sided neglect. AB - Egocentric models of neglect explain the lateralised omission of stimuli in neglect patients by an ipsilesional shift of a subjective reference frame. However, they differ in the direction of shift (rotation around the midsagittal plane versus translation in front/back space). We tested this hypothesis in a patient (AJ) with persistent right-sided neglect following a left temporo-parieto occipital and hypoxic lesion and in six age-matched healthy subjects. AJ showed visual neglect in line bisection, size matching, reading and visual search. Auditory localization was tested by using two different psychophysical techniques based on binaurally simulated stimuli for the horizontal plane in front and back space. Eye position was continuously monitored during stimulus presentation in all subjects. AJ revealed a significant ipsilesional, leftward shift of his auditory subjective median plane (ASMP) in front space (mean: -22.6 degrees), and a rightward shift of the ASMP in back space (+14.5 degrees). This pattern of results was replicated with a different psychophysical technique in a retest 10 months later. The rotational shift of AJ's ASMP contrasted with normal performance in the healthy subjects. Monaural hearing deficits can not account for these differential findings as all subjects (including AJ) performed normally. In conclusion, a rotation of the egocentric spatial reference frame may occur in the auditory modality for right-sided neglect. PMID- 16256150 TI - Mlh1-dependent suppression of specific mutations induced in vivo by the food borne carcinogen 2-amino-1-methyl-6-phenylimidazo [4,5-b] pyridine (PhIP). AB - Disruption of the DNA mismatch repair (MMR) pathway results in elevated mutation rates, inappropriate survival of cells bearing DNA damage, and increased cancer risk. Relatively little is known about the potential impact of environmentally relevant carcinogens on cancer risk in individuals with MMR-deficiency. We determined the effect of MMR status (Mlh1+/+ versus Mlh1-/-) on mutagenesis induced by the cooked-meat mutagen, 2-amino-1-methyl-6-phenylimidazo [4,5-b] pyridine (PhIP) within cII and supFG1 transgene reporters. Despite being a lymphomagen in mice, PhIP was not mutagenic in thymus. In colon, PhIP exposure induced 3-fold more mutations in Mlh1-deficient mice compared to their Mlh1+/+ littermates. Similar induction was seen in Mlh1-/- small intestine. Analysis of mutational spectra revealed that G/C to T/A transversions, the "signature PhIP mutation", were induced to similar levels regardless of Mlh1 status. In contrast, Mlh1-/- mice exhibited hypermutability to frameshifts, G/C to A/T transitions, and G/C to C/G transversions. Thus, both the level and types of mutation induced by PhIP are influenced by the activity of the MMR system. MMR may suppress PhIP induced mutation through recognition and processing of specific mispairs (PhIP G/T, PhIP-G/G, and PhIP-G/loop mispairs). In contrast, the PhIP-G/A mispair is unlikely to be a MMR substrate. In addition, the similar induction of both transversions and transitions in Mlh1-/- mice suggests that mutagenic bypass of PhIP-G is similarly efficient with dATP, dTTP, and dGTP, in contrast to previously published conclusions. Our data suggests that MMR-deficiency would increase the likelihood of PhIP-induced carcinogenic mutations. Further evaluation of the risk that consumption of heterocyclic amines may impart to MMR deficient individuals therefore is warranted. PMID- 16256153 TI - Effects of the acoustic environment on learning in rats. AB - Although noise of moderate intensity is commonly present during experiments on animal learning and memory, its effects have usually not been considered. The present study investigated how differences in the acoustic environment affect learning and memory under otherwise identical conditions. Genetically defined rats learned a complex maze in noise of moderate intensity (70 dB) or in quiet. Noise had a profound effect on learning and behaviour. Noise-exposed rats made fewer errors, explored less and finished their trials sooner. Results show that the acoustic environment is an important variable in studies with animal models of learning and memory. This might explain why differences in learning and behavioural scores occur across laboratories even when the same rodent strains and apparently identical experimental protocols are used. As the effects probably rely on neurotransmitter systems common in vertebrates, similar effects can be expected in other species including humans. PMID- 16256154 TI - Peripheral triiodothyronine (T(3)) levels during escapable and inescapable footshock. AB - Changes in peripheral thyroid hormone levels are associated with changes in human affective disorders, particularly depression. In the current study we used an animal stress paradigm, proposed to be an animal model of depression, to examine peripheral T(3) levels during and after escapable or inescapable stress in adult male rats. In this model, one animal can control the termination of foot-shock stress by performing a lever press, and therefore experiences escapable stress. His lever press also terminates the shock for his yoked partner, who has no control over the stressor, and therefore experiences inescapable stress. In three separate experiments, blood samples were collected during and after one or two sessions of escapable/inescapable stress. We found that exposure to inescapable stress, but not escapable stress, caused a decrease in T(3) levels 120 min post stress initiation. Peripheral T(3) levels were not significantly altered in animals exposed to escapable stress. In sum, these results add to a large body of previous data indicating that psychological coping can prevent the effects of physical stress on many diverse systems. PMID- 16256155 TI - Higher accumulation of proteinase inhibitors in flowers than leaves and fruits as a possible basis for differential feeding preference of Helicoverpa armigera on tomato (Lycopersicon esculentum Mill, Cv. Dhanashree). AB - Tomato (Lycopersicon esculentum, Mill; cultivar- Dhanashree) proteinase inhibitors (PIs) were tested for their trypsin inhibitory (TI) and Helicoverpa armigera gut proteinases inhibitory (HGPI) activity in different organs of the tomato plants. Analysis of TI and HGPI distribution in various parts of the plant showed that flowers accumulated about 300 and 1000 times higher levels of TI while 700 and 400 times higher levels of HGPI as compared to those in leaves and fruits, respectively. Field observation that H. armigera larvae infest leaves and fruits but not the flowers could be at least partially attributed to the protective role-played by the higher levels of PIs in the flower tissue. Tomato PIs inhibited about 50-80% HGP activity of H. armigera larvae feeding on various host plants including tomato, of larvae exposed to non-host plant PIs and of various larval instars. Tomato PIs were found to be highly stable to insect proteinases wherein incubation of inhibitor with HGP even for 3h at optimum conditions did not affect inhibitory activity. Bioassay using H. armigera larvae fed on artificial diet containing tomato PIs revealed adverse effect on larval growth, pupae development, adult formation and fecundity. PMID- 16256156 TI - Biological and molecular characterization of a canine hemangiosarcoma-derived cell line. AB - Canine hemangiosarcoma (HSA) is a devastating disease. Investigation of novel therapies has been limited by the limited availability of canine HSA-derived cell lines. We report the development of a canine HSA-derived cell line, DEN-HSA, which recapitulates features of angiogenic endothelium. DEN-HSA cells were derived from a spontaneous HSA arising in the kidney of a dog. DEN-HSA displayed surface molecules distinctive of endothelial histogenesis, including factor VIII related antigen, ICAM-1 and alpha(v)beta3 integrin. In vitro, DEN-HSA formed microvascular tube-like structures on Matrigel, and proliferated in response to a variety of angiogenic growth factors. The cells expressed mRNA and protein specific for bFGF and its receptors, and VEGF and its receptors, among others. DEN-HSA conditioned medium evoked a marked angiogenic response in a murine corneal pocket assay, indicating potent proangiogenic activity of substances secreted by this cell line. This research confirms the DEN-HSA cell line as endothelial in origin, suggests the presence of angiogenic growth factor autocrine loops, and offers the potential to utilize DEN-HSA cells for the study of novel therapies that modulate endothelial proliferation. PMID- 16256152 TI - A neural systems analysis of the potentiation of feeding by conditioned stimuli. AB - Associative learning processes play many important roles in the control of food consumption. Although these processes can complement regulatory mechanisms in the control of eating by providing opportunities for the anticipation of upcoming needs, they may also contribute to inappropriate or pathological consumption patterns by overriding internal regulatory signals. In this article, we first review some of the ways in which associative learning can contribute to the control of feeding, and then describe a neural systems analysis of a simple animal model of the control of feeding by Pavlovian-conditioned stimuli (CSs). Food-sated rats increase their food consumption after presentation of CSs that were previously paired with food while the rats were food-deprived. This cue potentiated feeding is independent of conditioned approach responses, and is at least somewhat specific to the foods associated with those CSs. A series of studies that used neuroanatomical tract tracing, immediate early gene expression, and neurotoxic disconnection lesion techniques implicated circuitry that includes the basolateral complex of the amygdala, the lateral hypothalamus, and the medial prefrontal cortex, but not the amygdala central nucleus, nucleus accumbens, or lateral orbitofrontal cortex, in cue-potentiated feeding. These studies also showed dissociations between cue-potentiated feeding and other learned motivational phenomena that are known to depend on function of amygdala systems. The data suggest that cue-potentiated feeding is uniquely mediated by cortical and amygdalar neurons that directly target the lateral hypothalamus, and thus gain access to hypothalamic neuropeptide and other systems involved in the promotion and suppression of eating. PMID- 16256157 TI - Steroidal glycosides from Cynanchum forrestii Schlechter. AB - Nine new steroidal glycosides, cynaforrosides B, C, D, E, and F, based on a 13, 14: 14, 15-disecopregnane-type skeleton, cynaforrosides G, H, and I with a new aglycone named cynaforrogenin A, and cynaforroside J together with three known C21 steroidal glycosides cynatratoside A, hancoside and komaroside C were isolated from the ethanol extract of the roots of Cynanchum forrestii Schlechter. The structures of new compounds were determined on the basis of spectral and chemical evidence. Steroidal glycosides with three kinds of skeletons were isolated from this plant simultaneously. The sugar units of cynaforrosides B-I contained two moieties of glucoses and especially cynaforrosides E-I contained two glucoses with the mode of 1-->6 linkage, which were rare among steroidal glycosides of the genus Cynanchum. PMID- 16256158 TI - A semi-symmetric two-locus model. AB - The two-locus symmetric viability model characterized by its invariance with respect to the exchange of alleles at each locus, is a well-studied model of classical two-locus theory. The symmetric model introduced by Lewontin and Kojima is among the few multi-locus models with epistatic interactions between loci for which a polymorphism with linkage equilibrium can be stable and this happens when recombination is sufficiently large. We show that an analogous property holds true for a different model, in which symmetry need exist at only one locus. The properties of this new semi-symmetric model are compared with those of the classical symmetric model. For tight linkage, two classes of polymorphisms are possible, depending on the magnitude of additive epistasis. The recombination rate above which linkage equilibrium becomes stable is derived analytically. As in the symmetric model, intervals of recombination in which no polymorphism is stable are possible, and stable polymorphisms can coexist with stable fixations. PMID- 16256159 TI - Toxins not neutralized by brown snake antivenom. AB - The Australian snakes of the genus Pseudonaja (dugite, gwardar and common brown) account for the majority of snake bite related deaths in Australia. Without antivenom treatment, the risk of mortality is significant. There is an accumulating body of evidence to suggest that the efficacy of the antivenom is limited. The current study investigates the protein constituents recognized by the antivenom using 2-DE, immuno-blot techniques and rat tracheal organ bath assays. The 2-DE profiles for all three snake venoms were similar, with major species visualized at 78-132 kDa, 32-45 kDa and 6-15 kDa. Proteins characterized by LC-MS/MS revealed a coagulant toxin ( approximately 42 kDa) and coagulant peptide ( approximately 6 kDa), as well as two PLA(2) ( approximately 14 kDa). Peptides isolated from approximately 78 kDa and 15-32 kDa protein components showed no similarity to known protein sequences. Protein recognition by the antivenom occurred predominantly for the higher molecular weight components with little recognition of 6-32 kDa MW species. The ability of antivenom to neutralize venom activity was also investigated using rat tracheal organ bath assays. The venoms of Pseudonaja affinis affinis and Pseudonaja nuchalis incited a sustained, significant contraction of the trachea. These contractions were attributed to PLA(2) enzymatic activity as pre-treatment with the PLA(2) inhibitor 4-BPB attenuated the venom-induced contractions. The venom of Pseudonaja textilis incited tracheal contractility through a non-PLA(2) enzymatic activity. Neither activity was attenuated by the antivenom treatment. These results represent the first proteomic investigation of the venoms from the snakes of the genus Pseudonaja, revealing a possible limitation of the brown snake antivenom in binding to the low MW protein components. PMID- 16256160 TI - Vitamin K1 (phylloquinone) induces vascular endothelial dysfunction: role of oxidative stress. AB - We aimed to investigate the mechanisms underlying the vascular effects induced by phylloquinone (Vitamin K1; VK1). Vascular reactivity experiments, using standard muscle bath procedures, showed that VK1 (5 and 50 microM) enhances the contractile response of endothelium-intact, but not denuded, rat carotid rings to phenylephrine. Similarly, maximal contraction induced by phenylephrine was enhanced in the presence of the nitric oxide (NO) synthase inhibitor NG-nitro-L arginine methyl ester (L-NAME). The combination of L-NAME and VK1 did not produce any further additional effect. Pre-incubation of intact-rings with VK1 reduced both acetylcholine- and bradykinin-induced relaxation. VK1 induced an increment in tension on carotid rings submaximally pre-contracted with phenylephrine. VK1 induced increment in tension was completely abolished by endothelial removal or incubation of intact rings with L-NAME and L-NNA. Conversely, 7-nitroindazole, 1400 W, or indomethacin did not affect VK1-induced contraction. Moreover, VK1 reduced L-arginine-induced relaxation in endothelium-intact rings. Lucigenin amplified chemiluminescence assays showed that VK1 induced an increase in the level of superoxide anions in endothelium-intact but not denuded rings. Measurement of nitrite and nitrate generation showed that VK1 did not alter nitrate formation but strongly inhibited the generation of nitrite. Finally, the superoxide anions scavenger tiron prevented the endothelial vasomotor dysfunction caused by VK1 on phenyleprine-induced contraction and acetylcholine or bradykinin induced relaxation. In conclusion, our data show that VK1 disrupts the vasomotor function of rat carotid. Our results suggest that VK1-induced oxidative stress through production of superoxide anion is interfering with the NO pathway, which in turn is responsible for the altered vascular reactivity induced by VK1. PMID- 16256161 TI - Immunization with a single extracellular enveloped virus protein produced in bacteria provides partial protection from a lethal orthopoxvirus infection in a natural host. AB - Subunit vaccines that use the vaccinia virus extracellular envelope protein A33R alone or combined with other structural proteins are excellent candidates for a new smallpox vaccine. Since a new smallpox vaccine would be used in humans, who are the natural hosts for the Orthopoxvirus variola, the agent of smallpox, it would be important to determine whether a prospective smallpox vaccine can protect from a lethal Orthopoxvirus infection in a natural host. We addressed this question using the mouse-specific Orthopoxvirus ectromelia virus. We demonstrate that immunization with recombinant ectromelia virus envelope protein EVM135 or its ortholog vaccinia virus A33R produced in E. coli protects susceptible mice from a lethal ectromelia virus infection. This is the first report that a subunit vaccine can provide protection to a lethal Orthopoxvirus infection in its natural host. PMID- 16256162 TI - Endemic versus epidemic viral spreads display distinct patterns of HTLV-2b replication. AB - As the replication pattern of leukemogenic PTLVs possesses a strong pathogenic impact, we investigated HTLV-2 replication in vivo in asymptomatic carriers belonging into 2 distinct populations infected by the same HTLV-2b subtype. They include epidemically infected American blood donors, in whom HTLV-2b has been present for only 30 years, and endemically infected Bakola Pygmies from Cameroon, characterized by a long viral endemicity (at least few generations). In blood donors, both the circulating proviral loads and the degree of infected cell proliferation were largely lower than those characterizing asymptomatic carriers infected with leukemogenic PTLVs (HTLV-1, STLV-1). This might contribute to explain the lack of known link between HTLV-2b infection and the development of malignancies in this population. In contrast, endemically infected individuals displayed high proviral loads resulting from the extensive proliferation of infected cells. The route and/or the duration of infection, viral genetic drift, host immune response, genetic background, co-infections or a combination thereof might have contributed to these differences between endemically and epidemically infected subjects. As the clonality pattern observed in endemically infected individuals is very reminiscent of that of leukemogenic PTLVs at the pre-leukemic stage, our results highlight the possible oncogenic effect of HTLV-2b infection in such population. PMID- 16256163 TI - Decreasing the frameshift efficiency translates into an equivalent reduction of the replication of the human immunodeficiency virus type 1. AB - The Gag-Pol polyprotein of the human immunodeficiency virus type 1 (HIV-1) is the precursor of the virus enzymatic activities and is produced via a programmed -1 translational frameshift. In this study, we altered the frameshift efficiency by introducing mutations within the slippery sequence and the frameshift stimulatory signal, the two elements that control the frameshift. These mutations decreased the frameshift efficiency to different degrees, ranging from approximately 0.3% to 70% of the wild-type efficiency. These values were mirrored by a reduced incorporation of Gag-Pol into virus-like particles, as assessed by a decrease in the reverse transcriptase activity associated to these particles. Analysis of Gag processing in infectious mutant virions revealed processing defects to various extents, with no clear correlation with frameshift decrease. Nevertheless, the observed frameshift reductions translated into equivalently reduced viral infectivity and replication kinetics. Our results show that even moderate variations in frameshift efficiency, as obtained with mutations in the frameshift stimulatory signal, reduce viral replication. Therapeutic targeting of this structure may therefore result in the attenuation of virus replication and in clinical benefit. PMID- 16256164 TI - Corneal topography, refractive state, and accommodation in harbor seals (Phoca vitulina). AB - Corneal topography of a harbor seal measured with a Placido's disc shows a central flattened stripe in the vertical meridian. Together with a pupil that can form a vertical slit, the flat vertical meridian can minimize the optical effects caused by the transition from water to air. Using infrared (IR) photoretinoscopy, we analyzed the refractive state of harbor seals and revealed a high degree of myopia and astigmatism in air, but emmetropia or slight hyperopia with little astigmatism underwater. The brightness distribution in the pupils suggest the presence of a multifocal dioptric apparatus in air and underwater. We found a first indication for accommodation by dynamic recordings underwater. PMID- 16256165 TI - Analysis of PDE6 function using chimeric PDE5/6 catalytic domains. AB - cGMP-phosphodiesterases of the PDE6 family are expressed in retinal photoreceptor cells, where they mediate the phototransduction cascade. A system for expression of PDE6 in vitro is lacking, thus straining progress in understanding the structure-function relationships of the photoreceptor enzyme. Here, we report generation and characterization of bacterially expressed chimeric PDE5/6 catalytic domains which are highly soluble, catalytically active, and sensitive to inhibition by the PDE6 Pgamma subunit. Two flexible PDE6 loops, H and M, impart chimeric PDE5/6 catalytic domains with PDE6-like properties. The replacement of the PDE6 H-loop into the PDE5 catalytic domain increases the catalytic rate and the K(m) value for cGMP hydrolysis, whereas the substitution of the M-loop produces catalytic PDE domains responsive to Pgamma. Multiple PDE6 segments preventing functional expression of the catalytic domain are identified, supporting the requirement for specialized photoreceptor chaperones to assist PDE6 folding in vivo. PMID- 16256166 TI - The effects of opposite-polarity dipoles on the detection of Glass patterns. AB - Glass patterns--randomly positioned coherently orientated dipoles--create a strong sensation of oriented spatial structure. On the other hand, coherently oriented dipoles comprising dots of opposite polarity ("anti-Glass" patterns) have no distinct spatial structure and are very hard to distinguish from random noise. Although anti-Glass patterns have no obvious spatial structure themselves, their presence can destroy the structure created by Glass patterns. We measured the strength of this effect for both static and dynamic Glass patterns, and showed that anti-Glass patterns can raise thresholds for Glass patterns by a factor of 2-4, increasing with density. The dependence on density suggests that the interactions occur at a local level. When the Glass and anti-Glass dipoles were confined to alternate strips (in translational and circular Glass patterns), the detrimental effect occurred for stripe widths less than about 1.5 degrees, but had little effect for larger stripe widths, reinforcing the suggestion that the interaction occurred over a limited spatial extent. The extent of spatial interaction was much less than that for spatial summation of these patterns, at least 30 degrees under matched experimental conditions. The results suggest two stages of analysis for Glass patterns, an early stage of limited spatial extent where orientation is extracted, and a later stage that sums these orientation signals. PMID- 16256167 TI - Advanced oxidation effect of ozonation combined with electrolysis. AB - The advanced oxidation effect of ozonation combined with electrolysis (electrolysis-ozonation) was discussed through the treatment of 4-chlorobenzoic acid (4-CBA) as a hydroxyl radical probe. The mechanism of hydroxyl radical production by electrolysis-ozonation process was also estimated with a mathematical model. The experimental results revealed that the electrolysis ozonation process had a synergistic effect on the degradation of 4-CBA. The advanced oxidation effect of electrolysis-ozonation was inferred from standard potentials of relevant electrochemical reactions and mathematical model analysis to be mainly attributed to ()O(3)(-) promotion of O(3) at the cathodes. An increase in electric current improved the degradation rate of 4-CBA. However, the pseudo-first order degradation rate constant reached a plateau at high electric current densities, as ()O(3)(-) promotion of O(3) at the cathodes was regulated by O(3) transport process from the bulk to the cathodes in the range exceeding an electric current density of 10 Am(-2). Accordingly, the balance of O(3) transport flux and electric current is important for the efficient operation of the electrolysis-ozonation reactor. PMID- 16256168 TI - Generation of active entities by the pulsed arc electrohydraulic discharge system and application to removal of atrazine. AB - Reactions induced by the pulsed arc electrohydraulic discharge (PAED) system in aqueous solutions were studied. PAED was generated by a spark gap type power supply (0.5 kJ/pulse) with rod-to-rod type electrodes in water. The measurements of physical parameters showed that the discharge is characterized by a sudden drop of the voltage while a peak of current occurs. The pressure waveform is composed of a positive pressure wave (shock wave) followed by negative pressure waves (expansion waves with a multiple wall reflection wave). The optical emission arc spectrum covers the UV-B, UV-A and visible zone with a maximum intensity in the range 380-425 nm. Peaks were representative of OH() radicals and atomic hydrogen emission lines. The identification of typical by-products from the removal of selected compounds in aqueous solution showed that PAED is the origin of photolysis, oxidation and reduction reactions. The impact of scavengers for OH() radicals or solvated electrons on the removal of atrazine and the concentration of the by-product deethylatrazine allowed the study of the combined and separate effects of the active entities. The energy efficiency of the PAED system can be improved by varying the gap of the electrodes in water. PMID- 16256169 TI - Biological removal of polychlorinated dibenzo-p-dioxins from incinerator fly ash by Sphingomonas wittichii RW1. AB - The ability of Sphingomonas wittichii strain RW1 to remove polychlorinated dibenzo-p-dioxins (PCDDs) from fly ash was investigated. All experiments were carried out in a slurry-phase system. Preliminary studies with resting cells of strain RW1 in a model fly ash system showed the complete removal of dibenzofuran (DF) and 81% of dibenzo-p-dioxin (DD). Incubation of real fly ash collected from municipal waste incinerators with strain RW1 for 15 days resulted in a 75.5% reduction in toxic PCDDs. When the same experiment was carried out using dead strain RW1 cells a 20.2% reduction in toxic PCDDs was observed, indicating that adsorption onto biomass was an important factor in dioxin elimination. Further analyses revealed that live strain RW1 cells removed 83.8% of the 2,3,7,8 substituted congeners from the fly ash, while dead cells removed 32.1% of the same congeners. To enhance the removal efficiency of toxic PCDDs, the effects of adding surfactant, repeated inoculation, and pre-adaptation of cultures were also studied. The removal of toxic PCDDs was enhanced by up to 10.3% upon repeated inoculation of the strain RW1, but was not much affected by the addition of surfactant. The present results suggest that S. wittichii strain RW1 is a potential candidate for the industrial removal of PCDDs from incinerator fly ash. PMID- 16256171 TI - Drosophila diet restriction in practice: do flies consume fewer nutrients? AB - Although many studies of Drosophila melanogaster report an effect of diet upon lifespan, it is assumed rather than know that adults maintained on restricted diet acquire fewer nutrients. Diet is restricted in practice by feeding flies a medium where nutrients are diluted and some early reports suggest that flies on diluted medium compensate with increased food intake to consume equal calories on all diets. Here we measure the feeding rate of adult Drosophila upon a yeast restricted diet that increases survival and reduces fecundity. We directly assessed food intake from the volume of consumed dye-marked medium and from the quantity of marked fecal pellets. Females were longest lived on diet with intermediate yeast concentration but food intake was greatest on diet with abundant yeast. Rather than compensatory feeding upon diluted diet, females increased food intake on the diet with elevated nutrient concentration. Consequently, females on diluted yeast-limited media will consume fewer calories as well as less yeast. To understand the importance of specific nutrients relative to calories as mediators of Drosophila aging, we must directly measure food intake and control for the feeding stimulation of nutrients. PMID- 16256170 TI - Dental development in Megaladapis edwardsi (Primates, Lemuriformes): implications for understanding life history variation in subfossil lemurs. AB - Teeth grow incrementally and preserve within them a record of that incremental growth in the form of microscopic growth lines. Studying dental development in extinct and extant primates, and its relationship to adult brain and body size as well as other life history and ecological parameters (e.g., diet, somatic growth rates, gestation length, age at weaning), holds the potential to yield unparalleled insights into the life history profiles of fossil primates. Here, we address the absolute pace of dental development in Megaladapis edwardsi, a giant extinct lemur of Madagascar. By examining the microstructure of the first and developing second molars in a juvenile individual, we establish a chronology of molar crown development for this specimen (M1 CFT = 1.04 years; M2 CFT = 1.42 years) and determine its age at death (1.39 years). Microstructural data on prenatal M1 crown formation time allow us to calculate a minimum gestation length of 0.54 years for this species. Postnatal crown and root formation data allow us to estimate its age at M1 emergence (approximately 0.9 years) and to establish a minimum age for M2 emergence (>1.39 years). Finally, using reconstructions or estimates (drawn elsewhere) of adult body mass, brain size, and diet in Megaladapis, as well as the eruption sequence of its permanent teeth, we explore the efficacy of these variables in predicting the absolute pace of dental development in this fossil species. We test competing explanations of variation in crown formation timing across the order Primates. Brain size is the best single predictor of crown formation time in primates, but other variables help to explain the variation. PMID- 16256172 TI - Arsenic concentrations in Chinese coals. AB - The arsenic concentrations in 297 coal samples were collected from the main coal mines of 26 provinces in China were determined by molybdenum blue coloration method. These samples were collected from coals that vary widely in coal rank and coal-forming periods from the five main coal-bearing regions in China. Arsenic content in Chinese coals range between 0.24 to 71 mg/kg. The mean of the concentration of Arsenic is 6.4+/-0.5 mg/kg and the geometric mean is 4.0+/-8.5 mg/kg. The level of arsenic in China is higher in northeastern and southern provinces, but lower in northwestern provinces. The relationship between arsenic content and coal-forming period, coal rank is studied. It was observed that the arsenic contents decreases with coal rank in the order: Tertiary>Early Jurassic>Late Triassic>Late Jurassic>Middle Jurassic>Late Permian>Early Carboniferous>Middle Carboniferous>Late Carboniferous>Early Permian; It was also noted that the arsenic contents decrease in the order: Subbituminous>Anthracite>Bituminous. However, compared with the geological characteristics of coal forming region, coal rank and coal-forming period have little effect on the concentration of arsenic in Chinese coal. The average arsenic concentration of Chinese coal is lower than that of the whole world. The health problems in China derived from in coal (arsenism) are due largely to poor local life-style practices in cooking and home heating with coal rather than to high arsenic contents in the coal. PMID- 16256174 TI - Lactate production by thrombin-activated platelets of patients with primary thrombocythemia. AB - INTRODUCTION: Platelet activation needs a high energy demand which is supplied by the degradation of glucose into lactate. Platelet response to agonists in patients with primary thrombocythemia is defective. We studied the production of lactate by the platelets of patients with this disease and defective platelet aggregation. MATERIAL AND METHODS: Ten patients suffering from primary thrombocythemia and ten controls were included in this study. The lactate generation was measured in resting and thrombin activated platelets in absence or presence of glucose. RESULTS: Resting platelets incubated for 30 min in phosphate buffered saline (PBS) generated the same amount of lactate in patients (44.6+/ 21.6 micromol/10(11) cells) and controls (41.0+/-17.3 micromol/10(11) cells). Addition of glucose led to similar increases in lactate formation by platelets in patients (82.2+/-26.4 micromol/10(11) cells) and controls (88.1+/-34.5 micromol/10(11) cells). The addition of thrombin in absence of glucose did not modify the lactate formation respective to PBS. Finally, the incubation of platelets with both glucose and thrombin caused further increases in the generation of lactate in both groups, patients (236.9+/-83.9 micromol/10(11) cells) and controls (228.6+/-63.5 micromol/10(11) cells) without differences between them. The production of lactate in both groups was also similar when platelets were incubated for 10 min or 20 min with both thrombin and glucose. However at 5 min, platelets of patients generated more lactate (97.8+/-23.7 micromol/10(11) cells) than controls (66.5+/-38.7 micromol/10(11) cells, p<0.05). CONCLUSIONS: These results suggest that thrombin is able to induce an initial hyperactivity of those pathways involved in the platelet energy production of patients with primary thrombocythemia. PMID- 16256173 TI - Kepone in James River fish: 1976-2002. AB - In late 1975, it was discovered that a manufacturing facility had not only exposed workers to the chlorinated pesticide, Kepone, but had also severely contaminated the James River estuary. To assess the potential for the public to be exposed to Kepone through the consumption of contaminated seafood, the Commonwealth of Virginia initiated a finfish-monitoring program in late 1975. Over 13,000 samples have been collected and analyzed as part of this effort. Kepone levels in most species began falling when the production of Kepone ended, but the average concentrations remained over the action limit of 0.3 microg g(-1) wet weight until the early 1980s. By 1988 few fish contained Kepone concentrations greater than the action limit. Kepone is still detected in the majority of white perch and striped bass samples taken from the James River and a fish consumption advisory is still in effect thirty years after the source of contamination was removed. PMID- 16256175 TI - The effect of vitamin A on CCl4-induced hepatic injuries in rats: a histochemical, immunohistochemical and ultrastructural study. AB - In this study, we aimed to investigate the effect of vitamin A on the transformation of the Ito cells to fibrogenic form and suppression of the development of fibrosis. Carbon tetrachloride intoxication was performed on rats for 2, 8, 12 or 20 weeks and 5x10(4) IU vitamin A (as retinol palmitate) was injected subcutaneously once every 4 weeks. Ito cells were detected by gold chloride impregnation, as well as desmin and alpha-smooth muscle actin (alpha SMA) immunohistochemistry. Additionally, all groups were examined ultrastructurally. The number of Ito cells that were labelled positively with gold impregnation decreased in the fibrotic groups; however, alpha-SMA and desmin immunopositive Ito cells increased. The samples from animals that were treated with vitamin A showed an increase in labelling with gold impregnation but a decrease in alpha-SMA immunopositivity. The data showed that vitamin A can prevent hepatic injury, by suppressing the transformation of Ito cells to fibrogenic form. We conclude that vitamin A has potential for the treatment of hepatic fibrotic diseases. Alpha-SMA immunohistochemistry was found to be more informative than desmin immunohistochemistry for monitoring liver fibrosis. PMID- 16256176 TI - Distribution of androgen receptor in rat ovarian follicles undergoing atresia at the beginning of pregnancy. AB - The immunohistochemical localisation of androgen receptor (AR) was investigated in a cohort of ovarian antral follicles developing, and subsequently undergoing atresia, in a hyperprolactinaemic milieu at the beginning of pregnancy in rats. Differentiation of the investigated follicles, observed during the first 5 days of pregnancy, was accompanied by a centripetal disappearance of androgen nuclear receptor in the granulosa layer, which did not include the cumulus oophorus complex and some antral granulosa cells. This pattern of decline resembled that typical of follicles maturing during the oestrous cycle but took longer to occur. The follicles did not ovulate and subsequently underwent atresia. The degeneration of some follicles was accompanied by a further loss of AR in the cumulus granulosa cells, but a strong positive AR immunoreaction persisted in the oocyte nucleoli. Some perinatal and early antral atretic follicles were found. In most cases their granulosa layers were AR-positive, although often only weakly. However, follicles with AR negative granulosa layers were also encountered. Nuclear immunolabelling for AR was positive in luteinized follicles. It can be concluded that follicular atresia involves changes in AR distribution which can be demonstrated as an abnormal depletion or persistence of AR. PMID- 16256177 TI - Pregranulomatous phase of sarcoidosis: immunohistochemical diagnosis. AB - Histopathological confirmation of clinical suspicion of sarcoidosis is based on the finding of non-caseating granulomas in biopsy material, usually in prescalene lymph nodes or in transbronchial lung biopsies. Lymph node reactive sinus histiocytosis (RSH) seen in relation to various inflammatory and non-inflammatory diseases can mimic the pregranulomatous phase of sarcoidosis (PSH). Differentiation of sinus histiocytosis based on histopathological features alone is limited. The purpose of this study is immunohistochemical determination of lymph node cellular response in granulomatous sarcoidosis, the PSH and RSH using a immunohistochemistry employing a panel of antibodies. Patient groups under study each contained 25 patients and included: those with clinical picture of sarcoidosis and non-caseating granulomatous lymphadenitis; those with confirmed sarcoidosis and with sinus histiocytosis without granuloma formation in lymph nodes; and finally, those without sarcoidosis and with "reactive" sinus histiocytosis in lymph nodes. Lymph node biopsy tissue was fixed in buffered formaldehyde, routinely processed to paraffin wax blocks, cut into 4-microm-thick sections, stained with hematoxylin and eosin and immunohistochemically labelled using a triple-layer APAAP protocol with purified polyclonal antibodies directed against SP 70 and SP90 from Mycobacterium tuberculosis and monoclonal antibodies against CD22, CD4, CD8, CD56, and CD68. Intensity of immunolabelling was assessed semiquantitatively by two independent observers. An increased CD4:CD8 ratio, moderate increase of immunolabelling for CD68 and slight decrease in immunolabelling for CD20, CD56, and SP90 was indicative of PSH when compared with RSH. The most notable difference between the studied groups was a difference in immunoreactivity to SP70 and CD4 antibodies. Lymph nodes with pregranulomatous sinus histiocytosis labelled with both antibodies. This profile of immunolabelling can be used in the differentiation of this condition from reactive sinusoidal lesions. PMID- 16256178 TI - B7-H4 overexpression in ovarian tumors. AB - OBJECTIVES: Despite great advances in therapeutic management, the mortality rate for ovarian cancer has remained relatively stable over the past 50 years. This study was designed to evaluate the expression of B7-H4 protein, recently identified as a potential molecular marker of breast and ovarian cancer by quantitative PCR analysis, in benign tumors, tumors of low malignant potential and malignant tumors of the ovary. METHODS: Archival formalin-fixed tissue blocks from serous, mucinous, endometrioid and clear cell ovarian tumors were evaluated by immunohistochemistry for the distribution of B7-H4 expression, and staining intensity was measured by automated image analysis. Univariate analyses were used to test for statistically significant relationships. RESULTS: B7-H4 cytoplasmic and membranous expression was detected in all primary serous (n = 32), endometrioid (n = 12), and clear cell carcinomas (n = 15), and in all metastatic serous (n = 23) and endometrioid (n = 7) ovarian carcinomas. By contrast, focal B7-H4 expression was detected in only 1/11 mucinous carcinomas. The proportion of positive cells and median staining intensity was greater in serous carcinomas than in serous cystadenomas or serous tumors of low malignant potential, and the differences were statistically significant (P < 0.0001 and P = 0.034, respectively). The median staining intensity was also significantly greater in endometrioid carcinomas than in endometriosis (P = 0.005). CONCLUSIONS: The consistent overexpression of B7-H4 in serous, endometrioid and clear cell ovarian carcinomas and the relative absence of expression in most normal somatic tissues indicates that B7-H4 should be further investigated as a potential diagnostic marker or therapeutic target for ovarian cancer. PMID- 16256179 TI - Consistent absence of BRAF mutations in cervical and endometrial cancer despite KRAS mutation status. AB - BACKGROUND: Mutational activation of KRAS and BRAF proto-oncogenes contributes to the development of many human cancers. Current research on gynecological cancer and specifically in cervical and endometrial cancer is focused on the mechanisms of their mutational activation. OBJECTIVES: In view of the paucity of data on their mutation frequency and the status of BRAF in these two types of gynecological cancer, we performed a systematic molecular study in 114 clinically and histologically well-defined malignant tumors of uterine cervix and endometrium and correlated the mutation status of KRAS and BRAF with the age at diagnosis and with tumor grade, stage or histological type. METHODS: Direct sequence analysis of the PCR products of KRAS and BRAF genes was used to screen for known activating mutations. RESULTS: In 67 cases of endometrial cancer, six KRAS mutations (8.9%) were found, four at codon 12 (5.9%) and two at codon 13 (2.9%), while no mutation was detected at codon 61. Most of the mutations occurred in surgical stage I and in the endometrioid adenocarcinoma subtype. We also detected three KRAS point mutations (6.3%) in the 47 cervical cancer samples, two at codon 12 (4.2%) and one at codon 13 (2.1%), while there was no mutation at codon 61. On the contrary, no mutation was identified in BRAF exon 15 for either endometrial or cervical cancer samples at position V600, which represents the most frequently mutated site of BRAF in human cancer. There was no association between KRAS mutations with either histological type, tumor grade or stage. Interestingly, however, KRAS mutation status in endometrial cancer was strongly associated with increased age at diagnosis (P < 0.001). CONCLUSIONS: Our data document (a) the absence of BRAF mutations in cervical and endometrial cancer, despite the mutation status of KRAS, (b) suggest that KRAS mutations reflect an early event in endometrial carcinogenesis and (c) imply that BRAF activation is involving alternative pathways in these two types of cancer. PMID- 16256180 TI - High prevalence of genital human papillomavirus type 52 and 58 infection in women attending gynecologic practitioners in South Taiwan. AB - BACKGROUND: We attempted to determine the prevalence of genital human papillomavirus (HPV) infection in women attending gynecologic practitioners in South Taiwan. METHODS: The population included 4383 women aged 16-78 seeking HPV testing at primary gynecologic practitioners regardless of their cervical cytology results. HPV DNA was identified from cervical swabs using semi-nested polymerase chain reaction with MY11, MY09/HMB01, and MY11/bioGP6+ primers. Genotyping for high-risk HPV (HR-HPV) was done separately by a HR-HPV chip, which contained 13 type-specific oligonucleotides on a nylon membrane. RESULTS: The overall HPV prevalence was 19.3% (849/4383), 11.1% (488/4383) were confirmed as HR-HPV positive. Among the women with HR-HPV infection, HPV-16 was the most prevalent type (22.1%; 108/488), followed by HPV-52 (21.3%; 104/488), and HPV-58 (19.9%; 97/488). Multiple infections were detected in 73 women (15.0%; 73/488). For women with age 30 or younger, the overall HPV and HR-HPV prevalence were 32.0% and 20.7%, respectively, which were significantly higher than those of women age older than 30 (17.2% and 9.5%, P < 0.001). More multiple infections (22.1% vs. 12.4%) were also found in women with age 30 or younger (P = 0.021). However, the relative contribution of types to the overall HR-HPV positive among different age groups remains the same. CONCLUSIONS: Our results showed an HPV prevalence that is similar compared with worldwide levels. HPV prevalence and multiple infections rate were decreasing across the age groups. Unlike most previous studies, the relative high prevalence of HPV 52 and 58 among South Taiwan women has important implications in vaccine prophylaxis. PMID- 16256181 TI - Active coping mediates the association between religion/spirituality and quality of life in ovarian cancer. AB - OBJECTIVES: This study investigated the role of religion/spirituality (R/S) and coping in quality of life (QOL) in 129 women immediately prior to a course of adjuvant chemotherapy for ovarian cancer. METHODS: Participants completed the COPE, the Functional Assessment of Cancer Therapy-Ovarian (FACT-O), and the Systems of Belief Inventory-15R (SBI-15R). Women averaged 58.9 years of age (SD = 11.5) and were primarily Caucasian (86%), married (74%), and had received at least some college education (67%). Eighty-five percent of the participants had stage III or IV ovarian cancer at study entry. RESULTS: Correlational analyses revealed that R/S was associated with active coping (r = 0.23, P = 0.022), overall QOL (r = 0.25, P = 0.012), emotional and functional well-being (r = 0.24, P = 0.014 and r = 0.28, P = 0.004), and fewer ovarian cancer-specific concerns (r = 0.27, P = 0.006). In addition, active coping was related to overall QOL (r = 0.22, P = 0.029) and social and functional well-being (r = 0.20, P = 0.042 and r = 0.33, P = 0.001). Tests of mediation between these variables suggested that the positive associations between R/S and functional well-being and R/S and overall QOL were mediated through the use of active coping. CONCLUSION: Future studies are needed to better understand the complex relationships between R/S, coping, and QOL throughout the ovarian cancer treatment experience. PMID- 16256182 TI - Juvenile granulosa cell tumor associated with pregnancy: Report of a case and review of the literature. AB - BACKGROUND: Juvenile granulosa cell tumors account for about 5% of all granulosa cell tumors and are diagnosed in nearly 80% of cases during the first two decades of life. Only 10% of granulosa cell tumors present during pregnancy. The incidence of ovarian malignancies during pregnancy varies from 0.05 to 0.07 per 1000 pregnancies. CASE: A 31-year-old pregnant woman was admitted to our university hospital due to an adnexal mass, 9.5 cm in diameter, which was detected at 34 weeks of gestation. At 37 + 5 weeks of gestation, a cesarean section with right salpingo-oophorectomy and removal of the tumor was performed. Histopathological findings, including immunohistochemical study, led to the diagnosis of juvenile granulosa cell tumor (JGCT). CONCLUSION: The histological features and the differential diagnosis of the JGCT are discussed. The optimal management of such adnexal masses during pregnancy is also discussed. A JGCT that is confined to the ovary appears to have an excellent prognosis and can be treated by unilateral salpingo-oophorectomy. PMID- 16256184 TI - Changing relationships of obesity and dyslipidemia in Greek children: 1982-2002. AB - BACKGROUND: The prevalence of childhood overweight and obesity has been increasing rapidly during the past decades in many parts of the world. Less is known with respect to the effects of increasing adiposity on blood lipid profile. The present study was designed to examine the influence of adiposity on secular trends in anthropometric characteristics and plasma lipids of Greek children. METHODS: A total of 419 and 374 boys with normal body weight and 109 and 246 boys with abnormal body weight (overweight and obese) were randomly recruited in 1982 and 2002, respectively (aged 12.1 +/- 0.1 years). Height, weight, body mass index (BMI, kg/m(2)), Rohrer index (kg/m(3)), plasma total cholesterol (TC), high density lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL C), triglyceride (TG), TC/HDL-C ratio, and LDL-C/HDL-C ratio were determined. RESULTS: Temporal changes in weight (positive), BMI (positive), HDL-C (negative), and cholesterol ratios (positive) were greater among overweight and obese vs. normal-weight boys (P < 0.05), while those for height (positive), LDL-C (positive), TG (positive), and TC (no change) were of similar magnitude. The increase in Rohrer index since 1982 failed to reach significance for children with normal body weight (P = 0.077) but did so for overweight and obese subjects (P = 0.027). CONCLUSIONS: These results indicate that secular increases in measures of fatness and adverse changes in plasma lipids were more pronounced among overweight and obese children than among normal-weight individuals, although qualitatively similar shifts were observed across the entire population. PMID- 16256183 TI - A multilevel ecological approach to promoting walking in rural communities. AB - BACKGROUND: Walking is a key focus of public health interventions yet is particularly uncommon in rural residents. This study's purpose was to determine whether a multilevel community intervention affected rates of moderate physical activity, in particular walking. METHODS: A quasi-experimental design examined changes in walking in six rural intervention communities in Missouri and six comparison communities in Arkansas and Tennessee in 2003-2004. Interventions were developed with community input and included individually tailored newsletters; interpersonal activities that stressed social support and health provider counseling; and community-wide events such as fun walks. A dose variable estimated exposure to intervention activities. Primary outcomes were rates of walking and moderate physical activity in the past week. RESULTS: At follow-up (n = 1531), the percentage of respondents who met the recommendation for walking was the same across the intervention and comparison areas. Among the dependent variables, walking showed some evidence of a positive linear trend across dose categories (P = 0.090). After adjusting for covariates and baseline rates, intervention participants in the moderate and high dose categories were about three times more likely to meet recommended guidelines for walking. CONCLUSIONS: Some evidence of effectiveness was shown for a multilevel intervention approach to promote walking. PMID- 16256185 TI - Anomalous mRNA levels of chromatin remodeling genes in swamp buffalo (Bubalus bubalis) cloned embryos. AB - The swamp buffalo (Bubalus bubalis) is a multi-purpose animal in agriculture that is challenged by extinction due to low reproductive efficiency. Nuclear transfer (NT) has been used to preserve special breeds of buffalo, as well as to increase the number of animals. However, cloned buffalo embryos have impaired development, as in other species. To understand the chromatin remodeling activities in cloned embryos and to improve NT technology, we examined the expression profiles of five genes involved in DNA and histone modifications, DNMT1, DNMT3A, DNMT3B, HAT1 and HDAC1, in single swamp buffalo metaphase II oocytes, NT and in vitro fertilized (IVF) embryos from the two-cell to the blastocyst stage, by quantitative real time RT-PCR. We observed similar expression dynamics for all genes studied in the NT and IVF embryos: relatively constant levels of expression for all genes were found from the MII oocyte up to the eight-cell stage; the levels of mRNA for HAT1 and DNMT3B continued to be stably expressed up to the blastocyst stage; while dramatic increases were seen for DNMT3A and HDAC1. Alternatively, the levels of DNMT1 started to decrease at the eight-cell stage. Despite the similarity in the dynamics of gene expression, dramatic differences in the relative levels of these genes between NT and IVF embryos were observed. The expression levels of all DNA modifying genes were higher in the NT embryos than in the IVF embryos at the eight-cell and blastocyst stages. The genes HDAC1 and HAT1 were also expressed significantly higher at the blastocyst stage in the NT embryos. Our results suggested differences in chromatin remodeling between NT and IVF embryos and that lower levels of DNA passive demethylation and higher levels of DNA de novo methylation occurred in the NT embryos. These observations are novel in the species of buffalo, and may be associated with developmental failure of cloned buffalo embryos due to the transcriptional repression effect of most genes studied here. PMID- 16256186 TI - Screening for high fertility in high-producing dairy cows. AB - A retrospective study involving 2756 pregnancies from two commercial dairy herds in northeastern Spain determined relationships between management, production and reproductive data, and high fertility (conception before 90 days in milk) in high producing dairy cows. High fertility was registered in 989 (35.9%) cows. The following data were recorded for each animal: herd, repeated animal (cows included two or more times within the study in which data were obtained from different lactational periods), parity (primiparous versus multiparous), previous twinning, reproductive disorders following calving (retained placenta, primary metritis) and at postpartum gynecological examination (incomplete uterine involution, pyometra and ovarian cysts), days in milk at conception, previous estrous synchronization and season of calving and conception. In order to evaluate the possible effect of high production during the peak milk yield on subsequent fertility, daily milk production at Day 50 postpartum was also recorded and cows were classified as high (> or = 50 kg) and low (< 50 kg) producers. Logistic regression analysis indicated no significant effects of herd, repeated animal, previous twinning, reproductive disorders such as primary metritis, incomplete uterine involution, pyometra and ovarian cysts, previous estrous synchronization and season of calving and insemination. Based on the odds ratio, the likelihood of high fertility increased in high-producer cows by a factor of 6.8. High fertility was less likely for multiparous cows (by a factor of 0.35) and for cows suffering placenta retention (by a factor of 0.65). High fertile cows produced a mean of 49.5 kg milk at Day 50 postpartum, in contrast to that 43.2 kg milk of the remainder cows. These findings question the negative effect of high production on fertility. Our results indicated that high individual cow milk production can be positively related to high fertility. PMID- 16256187 TI - Comparison between different markers for sperm quality in the cat: Diff-Quik as a simple optical technique to assess changes in the DNA of feline epididymal sperm. AB - The majority of wild felids, as well as some domestic cats, have low sperm concentration in their ejaculates, and a high proportion of abnormal spermatozoa. We have employed several possible semen quality markers to further characterize cat epididymal sperm. Methods included possible apoptotic reporters, such as the annexin V assay to monitor exposure of phosphatidylserine (PS) on the outer leaflet of the plasma membrane, as well as cell integrity; and the TUNEL assay to quantify DNA breaks. Sperm surface ubiquitination, another putative marker of sperm quality, was also monitored. The annexin V assay revealed a high percentage of sperm with PS exposure, and the TUNEL assay pointed to high levels (13+/-12%) of sperm with DNA breaks. Correlations were found between apoptotic markers (but not ubiquitination) and semen parameters. In parallel to this analysis, cat sperm morphology was evaluated using the Diff-Quik optical stain, which has been used in human reproduction laboratories. Several types of abnormalities could be characterized with this method. Remarkably, head staining abnormalities detected using the Diff-Quik staining method were strongly correlated with, and could accurately predict, sperm DNA defects detected in the same sample using the TUNEL assay. We therefore suggest that sperm morphology analysis using Diff-Quik could be used in field conditions to assess sperm status, due to the simplicity of the procedure and the equipment involved. PMID- 16256188 TI - Fertility in South Australian commercial Merino flocks: aspects of management. AB - Wide variation in reproductive performance of commercial Merino flocks in south central Australia is the result of genetic and environmental influences that are both amenable to change through decisions of management. Relationships of reproductive traits (estrus, ovulation, fertility, fecundity, lamb survival, and lambs weaned) with variables that graziers can change or modify (strain of Merino, day or month of exposure of ewes to rams, ram effect or teasing, length of the mating period, ram percentage, days between weaning and next mating, stocking density and flock size at lambing, ewe liveweight, and condition) are reported in this paper, the third in a series. Small differences were observed between medium and strong-wool South Australian Merino strains for reproductive traits. Choosing the time of year that ewes are exposed to rams, between late spring to autumn, may result in reduced ovulation rate during early summer (December) giving a potentially smaller net reproductive efficiency (lambs weaned). The ram effect or teasing, used by about 50% of graziers to synchronise lambing, could be effectively employed to the end of January. The technique was not reproductively advantageous when compared with flocks that were not teased. The percentage of rams mated to ewes varied widely (approximately 1-3%) and did not alter flock fertility, suggesting that a substantial proportion of graziers could safely reduce the number of rams purchased. A positive relationship between incidence of estrus during the first 14 d of the cycle and the number of days from weaning to next mating and a negative relationship of returns to service with the same variable indicates that managers should consider increasing the time allowed for recovery of liveweight and body condition by adjusting age at weaning, length of the mating period, or both. Lamb survival was curvilinearly related to flock size and not stocking intensity, with the optimum size at about 400 ewes. The number of lambs weaned per 100 ewes exposed to rams increased by 1.0 kg(-1) increase in liveweight at mating. We concluded that the major factor controlling net reproductive efficiency is nutritional in origin through its effects on ewe liveweight and condition, and is a factor that can be largely manipulated through management. PMID- 16256189 TI - Contribution of the oocyte to embryo quality. AB - The ability of a bovine embryo to develop to the blastocyst stage, to implant and to generate a healthy offspring is not a simple process. To clarify the importance of the contribution of the oocyte to the embryo quality, it is important to define more precisely the different types of competence expressed by oocytes. The ability to resume meiosis, to cleave upon fertilization to develop into a blastocyst, to induce pregnancy and to generate an healthy offspring are all separate events and succeeding in the first events does not ensure the success of subsequent ones. Furthermore, these events are associated with the three types of maturation processes observed in the oocyte: meiotic, cytoplasmic and molecular. These abilities vary also upon the type of follicle the oocytes is removed from. Larger or slow-growing follicles have been shown to foster better eggs than small or actively growing follicles. Hormonal stimulation can also affect oocyte competence with the nature of the effect (positive or negative) depending on timing and dose. This complex situation requires better definition of the contribution of each factor affecting the oocyte competence and the resulting embryo quality. PMID- 16256190 TI - Application of satellite infrared data for mapping of thermal plume contamination in coastal ecosystem of Korea. AB - The 5900 MW Younggwang nuclear power station on the west coast of Korea discharges warm water affecting coastal ecology [KORDI report (2003). Wide area observation of the impact of the operation of Younggwang nuclear power plant 5 and 6, No. BSPI 319-00-1426-3, KORDI, Seoul, Korea]. Here the spatial and temporal characteristics of the thermal plume signature of warm water are reported from a time series (1985-2003) of space-borne, thermal infrared data from Landsat and National Oceanic and Atmospheric Administration (NOAA) satellites. Sea surface temperature (SST) were characterized using advanced very high resolution radiometer data from the NOAA satellites. These data demonstrated the general pattern and extension of the thermal plume signature in the Younggwang coastal areas. In contrast, the analysis of SST from thematic mapper data using the Landsat-5 and 7 satellites provided enhanced information about the plume shape, dimension and direction of dispersion in these waters. The thermal plume signature was detected from 70 to 100 km to the south of the discharge during the summer monsoon and 50 to 70 km to the northwest during the winter monsoon. The mean detected plume temperature was 28 degrees C in summer and 12 degrees C in winter. The DeltaT varied from 2 to 4 degrees C in winter and 2 degrees C in summer. These values are lower than the re-circulating water temperature (6-9 degrees C). In addition the temperature difference between tidal flats and offshore (SSTtidal flats - SSToffsore) was found to vary from 5.4 to 8.5 degrees C during the flood tides and 3.5 degrees C during the ebb tide. The data also suggest that water heated by direct solar radiation on the tidal flats during the flood tides might have been transported offshore during the ebb tide. Based on these results we suggest that there is an urgent need to protect the health of Younggwang coastal marine ecosystem from the severe thermal impact by the large quantity of warm water discharged from the Younggwang nuclear power plant. PMID- 16256192 TI - The uptake of titanium ions by hydroxyapatite particles-structural changes and possible mechanisms. AB - In order to understand the effect of titanium ions on the molecular structure of hydroxyapatite (HAp), HAp powders were incubated in solutions with different titanium concentrations. After incubation, the powders obtained were analysed using different techniques, namely X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FT-IR), differential thermal analysis (DTA), X-ray photoelectron spectroscopy (XPS), and energy dispersive spectroscopy (EDS). The results suggest that, depending on the concentration of titanium in solution, two different mechanisms of interaction with HAp occur. For concentrations equal to or smaller than 200 ppm, the titanium uptake by the solid seems to be primarily due to incorporation in the lattice. For higher concentrations, a dissolution precipitation process seems to occur, leading to formation of a titanium phosphate compound. PMID- 16256191 TI - Optimized use of a biodegradable polymer as a carrier material for the local delivery of recombinant human bone morphogenetic protein-2 (rhBMP-2). AB - To improve the efficacy of a block copolymer of poly-d, l-lactic acid with randomly inserted p-dioxanone and polyethylene glycol (PLA-DX-PEG) as a drug delivery system for recombinant human bone morphogenetic proteins (rhBMPs), we examined the relationship between the volume of PLA-DX-PEG, the dose of rhBMP-2 and osteoinduction in a mouse model of ectopic bone formation. In a series of studies, we compared the size and bone mineral content (BMC) of ectopically induced bone by PLA-DX-PEG and collagen sponges carrying different quantities of rhBMP (0, 1, 2, 5, 10, 20 microg). An additional experiment was designed to investigate how a range of PLA-DX-PEG polymer volumes (15, 30, 60, 90 mg) with a fixed rhBMP concentration (0.01 wt%), altered the size and BMC of the induced ossicle. The influence of polymer volume was also examined in a further experiment wherein a fixed amount of rhBMP was placed in a range of PLA-DX-PEG copolymer volumes to give different concentrations of the protein per implant (0.02-0.0017 wt%). The results indicate that the bone yields were linearly dependent on the dose of rhBMP and also were proportional to the polymer volume above the minimal concentration of rhBMP-2 (0.0017 wt% in this series). The optimal concentration of rhBMP-2 in PLA-DX-PEG was 0.003 wt% in mice. The data provide important insights into the fabrication of implants that provide efficacious delivery of rhBMP-2 using the lowest possible dose of this expensive osteoinductive protein. This information will be of value for the clinical use of BMPs. PMID- 16256193 TI - Regulation of transplacental water transfer: the role of fetoplacental venous tone. AB - We used the in vitro dually perfused human placental lobule to test the hypothesis that known vasoconstrictors of the fetal placental circulation, angiotensin II and the thromboxane mimetic U46619 could induce fetomaternal water transfer. Secondly, we used a combination of vasoconstrictor and mechanically induced increases in fetal placental circulatory pressure to examine the role of the venous system in this context. Fetal-side administration of angiotensin II (A II) and U46619 (n=6 and n=9, for A-II and U46619, respectively) induced dose dependent, recoverable elevations in fetal inflow hydrostatic pressure (HP; A-II: maximum contractility=83 mmHg, EC50=22.0 nM; U46619: maximum contractility was not achieved, but exceeded the A-II effect) and loss of perfusate from the fetal side (A-II: EC50=70.2 nM, maximum fetal-side solvent loss=1906 microl/min; U46619: maximum fetal-side solvent loss was not achieved, but exceeded the A-II effect). Fetal-side solvent loss, for both agonists, was correlated linearly with fetomaternal inflow HP (FMIHP) in a biphasic manner (between 0 and 30 mmHg the slopes (+/-S.E.) were 6.4+/-2.2 and 17.1+/-5.8 microl/(min mmHg) for A-II and U46619, respectively; between 30 and 70 mmHg the slopes (+/-S.E.) were 35.6+/-6.5 and 43.7+/-15.9 microl/(min mmHg) for A-II and U46619, respectively). Increasing fetal-side lumenal pressure (n=3) by raising the fetal outflow catheter caused a loss of perfusate from the fetal side which was reduced in the presence of U46619 (fetal solvent loss per unit increase in fetal-side inflow HP: slopes were 1.198+/-0.123 and 0.783+/-0.085 microl/(min mmHg mmHg), respectively). Notwithstanding the possibility of fetoplacental arterial constriction, we conclude that vasoconstrictive agonists in the fetoplacental circulation affect venous resistance, causing fetomaternal fluid loss. These observations could be relevant to the oligohydramnios associated with intrauterine growth restriction, a condition associated with increased resistance in the umbilical circulation. PMID- 16256194 TI - Hypoxia stimulates Ca2+ release from intracellular stores in astrocytes via cyclic ADP ribose-mediated activation of ryanodine receptors. AB - The ability of O(2) levels to regulate Ca(2+) signalling in non-excitable cells is poorly understood, yet crucial to our understanding of Ca(2+)-dependent cell functions in physiological and pathological situations. Here, we demonstrate that hypoxia mobilizes Ca(2+) from an intracellular pool in primary cultures of cortical astrocytes. This pool can also be mobilized by bradykinin, which acts via phospholipase C and inositol trisphosphate production. By contrast, hypoxic Ca(2+) mobilization utilizes ryanodine receptors, which appear to be either present on the same intracellular pool, or on a separate but functionally coupled pool. Hypoxic activation of ryanodine receptors requires formation of cyclic ADP ribose, since hypoxic Ca(2+) mobilization was fully prevented by nicotinamide (which inhibits ADP ribosyl cyclase) or by 8-Br-cADP ribose, an antagonist of cyclic ADP ribose. Our results demonstrate for the first time the involvement of cyclic ADP ribose in hypoxic modulation of Ca(2+) signalling in the central nervous system, and suggest that this modulator of ryanodine receptors may play a key role in the function of astrocytes under conditions of fluctuating O(2) levels. PMID- 16256195 TI - Response of integrated biomarkers of fish (Lateolabrax japonicus) exposed to benzo[a]pyrene and sodium dodecylbenzene sulfonate. AB - Fish Lateolabrax japonicus were exposed to 0.1 and 1mg/L of anion surfactant sodium dodecylbenzene sulfonate (SDBS) and to 2 and 20 microg/L of benzo[a]pyrene (B[a]P) for 6, 12, and 18 days, with control and solvent control groups. Liver antioxidant enzymes, including superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), reduced glutathione (GSH), and glutathione S transferase (GST), were determined; brain acetylcholinesterase (AChE) and liver inducible nitric oxide synthase (iNOS) activities were also measured. The results indicated that (1) L. japonicus avoided oxidative damage through antioxidant systems; (2) SOD, GPx, and GSH were induced, and GST was inhibited and then induced by B[a]P exposure; and (3) CAT, GPx, and AChE were induced while iNOS was inhibited, and GST was induced and then inhibited by SDBS stress in experimental period. PMID- 16256196 TI - Ecotoxicity quantitative structure-activity relationships for alcohol ethoxylate mixtures based on substance-specific toxicity predictions. AB - Traditionally, ecotoxicity quantitative structure-activity relationships (QSARs) for alcohol ethoxylate (AE) surfactants have been developed by assigning the measured ecotoxicity for commercial products to the average structures (alkyl chain length and ethoxylate chain length) of these materials. Acute Daphnia magna toxicity tests for binary mixtures indicate that mixtures are more toxic than the individual AE substances corresponding with their average structures (due to the nonlinear relation of toxicity with structure). Consequently, the ecotoxicity value (expressed as effects concentration) attributed to the average structures that are used to develop the existing QSARs is expected to be too low. A new QSAR technique for complex substances, which interprets the mixture toxicity with regard to the "ethoxymers" distribution (i.e., the individual AE components) rather than the average structure, was developed. This new technique was then applied to develop new AE ecotoxicity QSARs for invertebrates, fish, and mesocosms. Despite the higher complexity, the fit and accuracy of the new QSARs are at least as good as those for the existing QSARs based on the same data set. As expected from typical ethoxymer distributions of commercial AEs, the new QSAR generally predicts less toxicity than the QSARs based on average structure. PMID- 16256197 TI - Is the 1:4 elutriation ratio reliable? Ecotoxicological comparison of four different sediment: water proportions. AB - Methodological research was carried out to evaluate the discriminatory capability of three toxicity bioassays toward different elutriation ratios (1:4, 1:20, 1:50, and 1:200 sediment:water ratios). Samples from six sampling stations of the Lagoon of Venice have been investigated. The toxicity bioassay results (sea urchin Paracentrotus lividus Lmk sperm cell and embryo toxicity bioassays and bivalve mollusk Crassostrea gigas Thunberg embryo toxicity bioassays) have shown that elutriates generated from the widely used 1:4 ratio were less toxic than those from intermediate ratios (1:20 and 1:50). PMID- 16256198 TI - Bacterial extracellular polymeric substance (EPS): a carrier of heavy metals in the marine food-chain. AB - The ecological implications of metal binding properties of bacterial EPS and its possible role in the bioaccumulation of pollutants in the marine food-chain was investigated using a partially purified and chemically characterized microbial EPS isolated from a species of Marinobacter. Various factors influencing metal sorption by the EPS including the influence of initial metal concentrations, incubation time, pH and sodium chloride concentrations on binding of lead (Pb2+) and copper (Cu2+) were evaluated. The bacterial EPS selectively bound more amount of Cu2+ per mg of EPS than Pb2+. Both copper and lead were sorbed more at near neutral pH than acidic pH. The sorption of Cu2+ increased with increasing copper concentration. The estimated maximum binding ability (MBA) of the EPS was 182 nmol copper and 13 nmol lead mg(-1) EPS. However, the sorption of these metals decreased with the increase in sodium chloride concentration. Furthermore, up to 35% of 14C-labeled Marinobacter was ingested by a benthic polychaete Hediste diversicolor. On an average, 29% of the ingested EPS was absorbed into tissues and 49% of the EPS was respired. It was apparent that the animals used the EPS as a source of energy and nutrition. The labile nature of the bacterial EPS and its ability to bind heavy metals might route the bound metals through the marine food chain, thereby transferring and aiding bioaccumulation of metal pollutants in the higher trophic animals. PMID- 16256199 TI - Identification of an NF-Y/HMG-I(Y)-binding site in the human IL-10 promoter. AB - The cis-acting elements and transcriptional factors that control interleukin 10 (IL-10) promoter activity remain to be clarified. In this report, by performing electrophoresis mobility shift assay (EMSA) with nuclear extracts from the Raji B cell line, the nuclear factor Y (NF-Y) was found to be capable of binding to a CCAAT box-containing region between the -215 and -192bps (the -215/-192 region), relative to the transcription initiation site, in the human IL-10 promoter. The binding of NF-Y to the promoter appears to be enhanced by the presence of HMG I(Y), since anti-HMG-I(Y) antibody diminished the NF-Y-binding activity in EMSA. Two T nucleotides at -192 and -193bps were found to be critical for the binding of NF-Y and HMG-I(Y) to the -215/-192 promoter region, suggesting the possibility that HMG-I(Y) binds to the AT-rich promoter region downstream of the CCAAT box at the -210/-199 region. These findings suggest that NF-Y and HMG-I(Y) may play an important role in regulating the IL-10 promoter activity in B cells. PMID- 16256200 TI - Protein kinase C-associated kinase is not required for the development of peripheral B lymphocyte populations. AB - Protein kinase C-associated kinase (PKK; DIK/RIP4) is an ankyrin-repeat containing serine/threonine receptor-interacting protein (RIP)-family kinase that can activate NFkappaB, and is required for keratinocyte development. In earlier studies, the expression of a catalytically inactive mutant of PKK in the B cell lineage resulted in a marked decrease in peripheral B cells in the spleen and a severe reduction of B-1 B cells. Here we explore the consequences of a null mutation in PKK with respect to the generation of peripheral B cell lineages and the activation of NFkappaB. We show that PKK is not required for the production of B cells in the bone marrow or for the development and maintenance of all mature B lymphocyte populations. We also show that PKK is not required for the activation of NFkappaB downstream of the BCR, CD40, or TLR-4 in B cells. Taken together, these data demonstrate that the loss of this RIP-family kinase does not compromise B lymphocyte development and maintenance, but leaves open the possibility that PKK may have a redundant role in these processes. PMID- 16256201 TI - Studies of interaction of dichloro[eta2-dimethyl-(2-methylidene-cyclohexylmethyl) amino]platinum(II) with DNA: effects on secondary and tertiary structures of DNA cytotoxic assays on human cancer cell lines Capan 1 and A431. AB - The interaction with DNA and the cytotoxic activity of a new organometallic platinum(II) compound were studied. Different techniques were used to evaluate changes in secondary and tertiary DNA structures, and to obtain images of DNA morphological changes. The ability of platinum complex to modify secondary DNA structure was explored by circular dichroism (CD). Electrophoretic mobility showed changes in tertiary DNA structure. Finally, Atomic Force Microscopy (AFM) revealed morphological changes of plasmid DNA (pBR322). This compound breaks the traditional structure-activity rules for cis-platinum compounds, but it could be of interest because of its different kinetics. An organometallic bond normally shows a higher trans-effect than an amine ligand, and that fact, a priori, could contribute to a higher DNA binding rate. Human A431 and Capan-1 cells (vulvae carcinoma and pancreatic carcinoma, respectively) were exposed to increasing concentrations of cisplatin and complex 6 for 24 h, after which time the cell number/viability was determined by the colorimetric MTT assay. A low cytotoxicity of organometallic compound 6 against A431 and Capan-1 cancer cell lines was observed and this result is consistent with the low interaction with DNA observed in previous studies. PMID- 16256202 TI - Structural and functional implications of the hexokinase-nickel interaction. AB - The interaction between nickel and yeast hexokinase was studied. The binding of nickel showed a positive cooperativity, and saturation was not reached. The nickel binding induced modifications in the secondary structure of the protein; thus, a lost of alpha helix and beta turns, as well as an increase of the random structure and beta sheet was observed. The monomer/dimmer equilibrium of the protein was modified in the presence of nickel, and the monomer state was mainly obtained at the highest nickel concentrations studied. These changes on the protein structure caused a decrease in the enzyme activity. According to kinetic studies, nickel caused a non-competitive inhibition when glucose was the variable substrate and a linear competitive inhibition when ATP was the variable substrate. PMID- 16256203 TI - Effects of UV-B in biological and chemical systems: equipment for wavelength dependence determination. AB - The thinning of the stratospheric ozone layer has prompted a large number of studies of UV-B-induced effects in biological and chemical systems. The wavelength dependency of such effects is of interest from mechanistic, physiological or economic points of view. Here, we describe an apparatus for determining the wavelength dependency of UV-B effects in biological and chemical systems. The apparatus consists of a high intensity UV radiation source and narrow bandpass filters to produce UV radiation in even intervals (between 280 and 360 nm). The usefulness of the equipment is demonstrated in two different systems: 1) Chalcone synthase (CHS) gene is up-regulated by UV-B radiation. Therefore quantitative analysis of the CHS gene expression was chosen in the present investigation for studies of the wavelength dependency of gene expression regulation in plants. Maximum induction of CHS expression was found at 300 nm with a 12-fold induction compared with the control; 2) The wavelength dependency of formation of dioxin-like photoproducts from the brominated flame retardant decabrominated diphenyl ether (DeBDE) is described. This is an example of UV-B induced conversion of non-toxic species into a number of products of which some may be toxic in the environment. In the UV interval studied, the highest dioxin like activity was found in the sample irradiated at 330 nm and therefore this wavelength is most important for the mechanism involved in photoconversion of DeBDE. PMID- 16256204 TI - Decoding emotional facial expressions in depressed and anorexic patients. AB - BACKGROUND: Several studies suggest the existence of impairments in the decoding of emotional facial expressions in various psychopathological conditions. This study investigates the recognition of emotional facial expressions (EFE) in young depressed patients and compares it to patients with eating disorders and control subjects. METHODS: 21 hospitalized female adolescents with major depression, 36 hospitalized female adolescents with eating disorders and 32 female control subjects were investigated with a set of emotional facial stimuli [Hess, U., Blairy, S., 1995. Set of Emotional Facial Stimuli, Department of Psychology, University of Quebec at Montreal, Montreal, Canada]. RESULTS: No significant differences were found between anorexic patients and controls in EFE decoding. Significant results were observed for depressed patients: they were less accurate in EFE decoding compared to anorexic patients and controls for the emotion of anger. LIMITATIONS: Previous studies on EFE decoding in depression and eating disorders used different facial stimuli or methodology. Furthermore, our study concerns only female subjects. These limitations could explain some discrepant result. CONCLUSIONS: The present results support the existence of impairments in the decoding of facial expressions in young female patients suffering from major depression. PMID- 16256205 TI - Suicidal behaviour in adolescents: associations with parental marital status and perceived parent-adolescent relationship. AB - BACKGROUND: Because equivocal findings exist with regard to the relationship between adolescents' suicidal behaviour and parental marital status, the aim of this study was to investigate this relationship and in particular the effect of the perceived parent-adolescent relationship on this association, taking into account the role of gender. METHOD: For this purpose, self-report surveys were administered to a representative school-based sample of 2707 adolescents in Antwerp (Belgium). RESULTS: 1) Boys living in a single parent family reported more suicidal ideations and self-harming behaviour than boys living in an intact family or in a remarried family; 2) Girls living in a remarried family reported more suicidal ideations and self-harming behaviour than girls living in an intact or in a single parent family; 3) Even after controlling for the levels of perceived parent-adolescent relationship, these associations remained significant. LIMITATIONS: The cross-sectional design, the retrospective assessment of suicidality and changes in family structure, the lack of external information and the assessment of the parent-adolescent relationship for both parents together, may have influenced the findings. CONCLUSIONS: When assessing risk factors for adolescent suicidality, marital status of the parents may bear clinical importance. In contrast to other studies, the perceived parent adolescent relationship did not alter this association, a finding that needs further study. PMID- 16256206 TI - Bovine monocytes and a macrophage cell line differ in their ability to phagocytose and support the intracellular survival of Mycobacterium avium subsp. paratuberculosis. AB - Bovine monocytes exhibited a greater ability to phagocytose Mycobacterium avium subsp. paratuberculosis (i.e. greater percentage of infected cells, and more bacilli per infected cell), than did a bovine macrophage cell line (BoMac). Phagocytosis of M. paratuberculosis by monocytes, but not the cell line, was significantly enhanced by the addition of autologous serum. Following ingestion, the numbers of viable M. paratuberculosis cells in monocytes increased during the first 4 days and then declined between day 4 and day 8 after infection, as determined by a radiometric method. In contrast, BoMac cells were not permissive for bacillary multiplication; the numbers of M. paratuberculosis remained largely unchanged in the cell line during the 8 day incubation period. The numbers of microscopically visible acid-fast bacilli increased with time in monocytes but not in the macrophage cell line. These observations suggest that replication and inactivation of bacilli may both occur in monocytes. The differing abilities of bovine monocytes and the macrophage cell line to ingest and restrain the intracellular growth of M. paratuberculosis provide contrasting model systems for investigating how M. paratuberculosis enters and persists within its preferred niche, the mononuclear phagocyte. PMID- 16256207 TI - Neonatal alcohol exposure increases malondialdehyde (MDA) and glutathione (GSH) levels in the developing cerebellum. AB - It has been suggested that developmental alcohol-induced brain damage is mediated through increases in oxidative stress. In this study, the concentrations of malondialdehyde (MDA) and reduced glutathione (GSH) were measured to indicate alcohol-mediated oxidative stress. In addition, the ability of two known antioxidants, melatonin (MEL) and lazaroid U-83836E (U), to attenuate alcohol induced oxidative stress was investigated. Sprague-Dawley rat pups were randomly assigned to six artificially-reared groups, ALC (alcohol), MEL, MEL/ALC, U, U/ALC, and GC (gastrostomy control), and one normal suckle control (to control for artificial-rearing effects on the dependent variables). The daily dosages for ALC, MEL, and U were 6 g/kg, 20 mg/kg, and 20 mg/kg, respectively. Alcohol was administered in 2 consecutive feedings, and antioxidant (MEL or U) was administered for a total of 4 consecutive feedings (2 feedings prior to and 2 feedings concurrently with alcohol). The animals received treatment from postnatal days (PD) 4 through 9. Cerebellar, hippocampal, and cortical samples were collected on PD 9 and analyzed for MDA and GSH content. The results indicated that MDA concentrations in the cerebellum were significantly elevated in animals receiving alcohol; however, MDA levels in the hippocampus and cortex were not affected by alcohol treatment. Additionally, GSH levels in the cerebellum were significantly elevated in groups receiving alcohol, regardless of antioxidant treatment. Neither antioxidant was able to protect against alcohol induced alterations of MDA or GSH. These findings suggest that alcohol might increase GSH levels indirectly as a compensatory mechanism designed to protect the brain from oxidative-stress-mediated insult. PMID- 16256208 TI - Imbalances emerge in cardiac autonomic cell signaling after neonatal exposure to terbutaline or chlorpyrifos, alone or in combination. AB - During early neonatal development, the future reactivity of the heart to cardiac autonomic stimulation is programmed by the timing and intensity of the arrival of parasympathetic and sympathetic inputs. In neonatal rats, we examined the effects of exposure to terbutaline, a beta-adrenoceptor (betaAR) agonist used to arrest preterm labor, and chlorpyrifos (CPF), a widely used organophosphate pesticide that acts in part through inhibition of cholinesterase, using scenarios mimicking the likely developmental stages corresponding to peak human exposures: postnatal days (PN) 2-5 for terbutaline and PN11-14 for CPF. Terbutaline evoked a progressive deficit in cardiac betaAR binding but did not interfere with the ability of the receptors to stimulate adenylyl cyclase (AC). Terbutaline also reduced expression of m2 muscarinic acetylcholine receptors and suppressed their ability to inhibit AC. Surprisingly, CPF produced similar actions, a decrement in betaAR and m2 muscarinic receptor binding and a loss of the cholinergic AC response, and also augmented the ability of betaARs to stimulate AC. The effects of CPF are thus unlikely to reside in cholinergic hyperstimulation resulting from cholinesterase inhibition but instead involve other actions converging on receptors and cell signaling. Exposure to both agents, terbutaline followed by CPF, produced a summation of the two individual effects. Our findings at the level of cell signaling thus indicate that neonatal exposure to terbutaline or CPF, or sequentially to both agents, results in an imbalance of cardiac autonomic inputs favoring increased excitability, an outcome that may have an impact on cardiovascular responses. PMID- 16256209 TI - In vitro Th2 deviation of myelin-specific peripheral blood lymphocytes from patients with multiple sclerosis. AB - This study aimed at investigating if selective ex vivo immune deviation of myelin specific cytokine secretion towards Th2 is possible in blood cells from patients with multiple sclerosis (MS). Interleukin (IL)-4 (Th2) and interferon-gamma (Th1) secreting cells were recorded by ELISPOT in 13 MS patients. Deviation was successful in 10 patients. Interleukin-4 alone was most effective in inducing myelin-specific immune deviation in MS patients whereas IL-1 or IL-15 in combination with IL-4 did not improve the results. Further studies and improvements are needed before ex vivo immune deviation can be considered a potential treatment in patients with MS. PMID- 16256210 TI - Interleukin-1alphabeta gene-deficient mice show reduced nociceptive sensitivity in models of inflammatory and neuropathic pain but not post-operative pain. AB - The pro-inflammatory cytokine interleukin-1 (IL-1) has been implicated in both inflammatory processes and nociceptive neurotransmission. To further investigate the role of IL-1 in different pain states, gene-disrupted mice lacking both IL 1alpha and IL-1beta genes (IL-1alphabeta (-/-)) were characterized in inflammatory, neuropathic, and post-operative pain models. IL-1alphabeta (-/-) mice showed normal sensorimotor function as measured by the rotorod assay compared to control mice (BALB/c). Acute and persistent formalin-induced nocifensive behaviors were reduced by 20% in IL-1alphabeta (-/-) mice as compared to control mice. IL-1alphabeta (-/-) mice also showed reduced inflammatory thermal and mechanical hyperalgesia compared to controls following the intraplantar administration of carrageenan or complete Freund's adjuvant (CFA). The duration of inflammatory hyperalgesia was shortened in IL-1alphabeta (-/-) mice versus controls in the CFA model. In contrast, deletion of IL-1alphabeta did not change the extent or the duration of post-operative pain developing after skin incision of the hind paw. Finally, time to onset, duration, and magnitude of mechanical allodynia were reduced in two models of neuropathic pain, spinal nerve L5-L6 ligation and chronic constriction injury of the sciatic nerve, in IL 1alphabeta (-/-) mice versus controls. These results demonstrate that IL 1alphabeta modulates both the generation and the maintenance of inflammatory and chronic neuropathic pain and that IL-1 may modulate nociceptive sensitivity to a greater extent in conditions of chronic as compared to acute pain. PMID- 16256211 TI - Protein synthesis and the mechanisms of lasting change in anxiety induced by severe stress. AB - Brief, unprotected exposure of rats to cats (predator stress) may be lastingly anxiogenic in a variety of tests of rodent anxiety. Recent findings suggest that predator stress induced plasticity in neural circuitry implicated in fear learning underlies some of these anxiogenic effects. In addition, recent work implicates a consolidation-like process in the impact of predator stress on anxiety in that effects of predator stress may be interrupted by immediate post stressor pharmacological interventions. The present study tested whether "consolidation" of the anxiogenic effects of predator stress were dependent on protein synthesis. In addition, the study examined whether a protein synthesis dependent reconsolidation-like process was at work when rats were exposed to a cat twice. Anisomycin (210 mg/kg) or vehicle (Tween 80 in saline) was injected subcutaneously 1 min after a single cat exposure (consolidation test paradigm) or a 1 min after a second cat exposure (reconsolidation test paradigm) and behavior tested 7-8 days after predator stress. In the consolidation test paradigm, anisomycin blocked the anxiogenic effects of predator stress in the elevated plus maze (EPM) measured with open arm exploration. Moreover, anisomycin blocked the potentiation of startle by predator stress when rats were startled in the light, but not when startled in the dark. In contrast, the delay of habituation of startle produced by predator stress was unaffected by anisomycin. Suppression of risk assessment in the EPM by predator stress was not affected by anisomycin either. In startle testing, vehicle injection 1 min after predator stress led to a lasting suppression, rather than enhancement of startle response. Vehicle plus predator stress enhanced and prolonged corticosterone level changes sampled over 30-180 min after treatment when compared to handled or predator stressed only rats. In addition, predator stress plus vehicle suppression of startle was blocked by a benzodiazepine anxiolytic (chloradiazepoxide) or the glucorticoid receptor (GR) blocker RU486. Both drugs returned startle to the predator stressed only heightened levels. It is argued that an added anxiogenic effect of vehicle injection plus predator stress leads to a suppression, rather than enhancement of startle. Startle suppression appears to be mediated, in part, by activation of GR by corticosterone which engages a protein synthesis dependent process, since anisomycin blocked the startle suppressive effects of vehicle. Startle suppression also appeared to be independent of the startle enhancing effect of predator stress and in competition with it. Since predator stress may model aspects of hyperarousal associated with post traumatic stress disorder (PTSD), implications of these findings for understanding of mechanisms of initiation of the disorder and for treatment are discussed. PMID- 16256212 TI - Social isolation alters the effect of desipramine in the rat forced swimming test. AB - The present study determined whether rearing in social isolation alters the behavioural response produced by the selective noradrenaline uptake inhibitor, desipramine when rats are exposed to the forced swimming test or an open field arena. Male Wistar rats were raised from weaning either alone (isolation rearing) or in groups of five or six rats/cage (social rearing) for 4 weeks before behavioural testing. The results demonstrated that sub-chronic administration of desipramine (5, 10 and 20 mg/kg i.p.) 24, 5 and 1 h caused a dose-related decrease in immobility time and increase struggling and produced locomotor hypoactivity compared with saline treated rats in both isolation and socially reared rats. However, these effects were significantly greater in isolation than socially reared rats. The results indicate isolation rearing alters the response to aversive stimuli and that sub-chronic antidepressant treatment potentiates these effects. PMID- 16256213 TI - Mice over-expressing the 5-HT(1A) receptor in cortex and dentate gyrus display exaggerated locomotor and hypothermic response to 8-OH-DPAT. AB - The serotonin 1A (5-HT(1A)) receptor is one of the best described receptor subtypes of the serotonergic system. Due to the complex distribution pattern, the pre- and postsynaptic localisation, the impact on various monoamines, as well as the influence on a wide range of physiological functions, the contribution of 5 HT(1A) receptors to behavioural outcomes is difficult to define. In this study, we present a new transgenic mouse model with a prominent over-expression of the 5 HT(1A) receptor in the outer cortical layers (I-III) and the dentate gyrus. Behavioural studies revealed a slight decrease in baseline motor activity of homozygous mice during the open field test. Moreover, core body temperature of male transgenic mice was significantly lower than that of wild-type mice. Pharmacological studies with the 5-HT(1A) receptor agonist 8-OH-DPAT (0.1-2.5 mg/kg, i.p.) revealed an exaggerated drug response in mutant mice. 8-OH-DPAT led to a drastic decrease in motor activity in the open field and elevated plus maze test. This significant effect on motor activity became more apparent by investigating the serotonergic syndrome induced by 8-OH-DPAT. Concentration as low as 0.5 mg/kg 8-OH-DPAT caused immobility in transgenic mice for 30 min, head weaving behaviour, and backward walking, whereas in wild-type animals, typical behaviours of the serotonin syndrome were first observed at concentrations of 1.5 mg/kg and more. In addition, the 8-OH-DPAT induced hypothermia was more pronounced in mutant mice than in wild-type animals. Therefore, these genetically modified mice represent a promising model for further investigations of the role of 5-HT(1A) receptors. PMID- 16256214 TI - Context is a trigger for relapse to alcohol. AB - The environment in which alcohol consumption occurs may trigger later relapse in alcohol abusers. In this study, we tested whether an alcohol-associated environment would induce alcohol-seeking behavior. Male rats were trained to lever press for oral alcohol reinforcement in a distinctive context. Responding was then extinguished in a context with different olfactory, visual and tactile properties. Placement of the rats back into the original context in which they self-administered alcohol induced, in the absence of alcohol availability, a significant increase in lever press responding on the alcohol lever as compared to extinction levels of responding. The ability of the alcohol context to support alcohol-seeking behavior was maintained over 3 weeks, with no significant diminution. A second group of rats was trained to lever press for sucrose reinforcement; this group also demonstrated context-dependent reinstatement, although the degree of reinstatement decreased over repeated tests, returning to extinction values after 3 weeks. These findings indicate that contextual conditioning has a long-term impact on ethanol-seeking behavior after ethanol withdrawal. This animal model may be useful to study the neural mechanisms underlying relapse induced by ethanol-associated contexts in humans. PMID- 16256215 TI - Echolocation, vocal learning, auditory localization and the relative size of the avian auditory midbrain nucleus (MLd). AB - The avian nucleus mesencephalicus lateralis, pars dorsalis (MLd) is an auditory midbrain nucleus that plays a significant role in a variety of acoustically mediated behaviours. We tested whether MLd is hypertrophied in species with auditory specializations: owls, the vocal learners and echolocaters. Using both conventional and phylogenetically corrected statistics, we find that the echolocating species have a marginally enlarged MLd, but it does not differ significantly from auditory generalists, such as pigeons, raptors and chickens. Similarly, all of the vocal learners tend to have relatively small MLds. Finally, MLd is significantly larger in owls compared to all other birds regardless of how the size of MLd is scaled. This enlargement is far more marked in asymmetrically eared owls than symmetrically eared owls. Variation in MLd size therefore appears to be correlated with some auditory specializations, but not others. Whether an auditory specialist possesses a hypertrophied MLd appears to be depend upon their hearing range and sensitivity as well as the ability to resolve small azimuthal and elevational angles when determining the location of a sound. As a result, the only group to possess a significantly large MLd consistently across our analyses is the owls. Unlike other birds surveyed, owls have a battery of peripheral and other central auditory system specializations that correlate well with their hearing abilities. The lack of differences among the generalists, vocal learners and echolocaters therefore reflects an overall similarity in hearing abilities, despite the specific life history requirements of each specialization and species. This correlation between the size of a neural structure and the sensitivity of a perceptual domain parallels a similar pattern in mammals. PMID- 16256216 TI - Adrenomedullin inhibits angiotensin II-induced contraction in human aortic smooth muscle cells. AB - The vasodilating peptide adrenomedullin (AM) has been reported to regulate vascular tone as well as proliferation and differentiation of various cell types in an autocrine/paracrine manner. Our study was designed to investigate the effect of AM on Ang II-induced contraction on human aortic smooth muscle cells (HASMC) in vitro, evaluating the signal pathways involved. Our findings indicate that AM was able to inhibit HASMC Ang II-induced contraction (IC50 19 nM). AM stimulated cAMP production in a dose-dependent fashion as well. SQ 22.536, an adenylate cyclase inhibitor, and KT5720, a PKA inhibitor, blunted the AM effect, suggesting that it was mediated by the activation of the cAMP transduction pathway. Our results suggest that AM plays a role in the regulation of HASMC contraction by antagonizing the Ang II effects and may be involved in conditions of altered regulation of the blood vessels. PMID- 16256217 TI - The impact of cognitive impairment on short-term outcomes of hip fracture patients. AB - The purpose of this study is to examine the differences in outcomes related to recovery after hip fracture among patients with and without cognitive impairment. This is a prospective cohort study of consecutively hospitalized elderly patients with acute hip fracture in a setting utilizing a multidisciplinary hip fracture service. Of the 48 patients admitted with hip fracture, 18 patients were found to have cognitive impairment postoperatively as determined by a Mini-Mental State Examination (MMSE) score 40 Gy (V40) to > 75 Gy (V75), and maximum esophageal doses. RESULTS: Sixteen patients developed grade 2-3 acute esophagitis (no G4 or 5). The maximum likelihood analysis produced new Lyman model parameters of: TD50 = 51 Gy, n = 0.44 and m = 0.32. The mean NTCP value is significantly lower (P < 0.001) in the group of patients without esophagitis (13.5%) than with esophagitis (27.2%). The rates of esophagitis are 2.5, 7, 9 and 13.4%, respectively, when the NTCP values are <10%, <15%, <20% and <25%. A significant association is found between esophagitis and dose/volume parameters V40 (P = 0.001) to V70 (P = 0.024). CONCLUSIONS: New values of TD50, n and m offer a good description of the esophagitis distribution in our population. Compared to the use of this model with previously published parameters (associated with late toxicity) predictions of the model for acute esophagitis using the new parameters would indicate that, for a population of patients, the distribution of events as a function of uniform dose would occur with a lower mean uniform dose value (smaller TD50), over a wider range of uniform doses (larger m), while also exhibiting a bigger volume effect (larger n). These new parameter values are supported in essence by the correlations found for the dose/volume threshold parameters. PMID- 16256231 TI - Observer variation in target volume delineation of lung cancer related to radiation oncologist-computer interaction: a 'Big Brother' evaluation. AB - BACKGROUND AND PURPOSE: To evaluate the process of target volume delineation in lung cancer for optimization of imaging, delineation protocol and delineation software. PATIENTS AND METHODS: Eleven radiation oncologists (observers) from five different institutions delineated the Gross Tumor Volume (GTV) including positive lymph nodes of 22 lung cancer patients (stages I-IIIB) on CT only. All radiation oncologist-computer interactions were recorded with a tool called 'Big Brother'. For each radiation oncologist and patient the following issues were analyzed: delineation time, number of delineated points and corrections, zoom levels, level and window (L/W) settings, CT slice changes, use of side windows (coronal and sagittal) and software button use. RESULTS: The mean delineation time per GTV was 16 min (SD 10 min). The mean delineation time for lymph node positive patients was on average 3 min larger (P = 0.02) than for lymph node negative patients. Many corrections (55%) were due to L/W change (e.g. delineating in mediastinum L/W and then correcting in lung L/W). For the lymph node region, a relatively large number of corrections was found (3.7 corr/cm2), indicating that it was difficult to delineate lymph nodes. For the tumor atelectasis region, a relative small number of corrections was found (1.0 corr/cm2), indicating that including or excluding atelectasis into the GTV was a clinical decision. Inappropriate use of L/W settings was frequently found (e.g. 46% of all delineated points in the tumor-lung region were delineated in mediastinum L/W settings). Despite a large observer variation in cranial and caudal direction of 0.72 cm (1 SD), the coronal and sagittal side windows were not used in 45 and 60% of the cases, respectively. For the more difficult cases, observer variation was smaller when the coronal and sagittal side windows were used. CONCLUSIONS: With the 'Big Brother' tool a method was developed to trace the delineation process. The differences between observers concerning the delineation style were large. This study led to recommendations on how to improve delineation accuracy by adapting the delineation protocol (guidelines for L/W use) and delineation software (double window with lung and mediastinum L/W settings at the same time, enforced use of coronal and sagittal views) and including FDG-PET information (lymph nodes and atelectasis). PMID- 16256232 TI - Optimization of follow-up timing from study of patterns of first failure after primary treatment. An example from patients with NSCLC: a study of the REACT working group of ESTRO. AB - BACKGROUND AND PURPOSE: The European Society for Therapeutic Radiology and Oncology was funded by the EU for a project on Recording providing Education, and Ameliorating the Consequences of Treatment (REACT). An important aim of follow-up (FU) after treatment for cancer is to detect various events associated with disease recurrence or metastatic spread or severe treatment-related complications as early as possible. Each tumour type may show a specific pattern and timing of these events related to different prognostic factors. The aim of this study was to propose a way of defining an optimal timing schedule for follow-up after treatment based on the analysis of failure patterns determined from follow-up data from prospective clinical trials. MATERIAL AND METHODS: Cox proportional hazards model was used to identify prognostic factors associated with each failure type (loco-regional recurrence (LR), distant metastasis (DM) or side effects (SE)). Competing risks methods were applied to estimate the cumulative incidence functions (CIF), adjusted on the significant prognostic factors. Equally spaced quantiles of the CIF were then used to estimate the corresponding optimised follow-up times depending on a pre-specified total number of visits. Follow-up data from the CHART bronchus clinical trial were used to analyse the pattern of time to first failure. RESULTS: A significantly higher risk of failure was observed for males (SE), stage III (DM) and conventional treatment (LR). Overall, patients treated with CHART needed 1 fewer visit in each category of patients compared to the Conventional group. For example, stage III male patients treated with CHART would need 8 visits during the first two years at 7, 11, 16, 24, 37, 52, 64 and 104 weeks rather than the 9 follow-up visits planned in the protocol. Similar patients treated with Conventional radiotherapy would need 8 visits at 3, 5, 7, 11, 15, 24, 52 and 104 weeks. CONCLUSIONS: Use of these methods would allow timing of follow-up visits to be adapted according to tumour site and prognostic factors determined previously from audit or clinical trials. Application of this approach could optimize the timing of follow-up visits by placing them closer to the times when failures are expected to occur. It does not address the wider issues of follow-up such as who should do it or what should be done for which further studies are required. PMID- 16256233 TI - Evidence that protease-activated receptor-2 mediates trypsin-induced reversal of stellation in cultured rat astrocytes. AB - Serine proteases such as thrombin and trypsin play a key role in the development and repair processes in the central nervous system. Molecular actions of serine proteases include multiple cellular events like activation of protease-activated receptors (PARs). PARs belong to a family of G protein-coupled receptors that can be stimulated through their proteolytic cleavage by ligands. PAR-2 has been implicated in neurodegenerative diseases including astrogliosis. Although recent studies have shown that low concentration of trypsin activates PAR-2, its role in morphological changes in primary astrocytes has not been studied. In the present study, we investigated the effects of PAR-2 in astrocyte stellation in rat primary astrocyte culture. Both trypsin (0.1-1 U/ml) and a PAR-2-activating peptide SLIGRL-NH2 (1-50 microM) significantly reversed the stellation induced by serum deprivation in rat astrocytes. Treatment of astrocytes with trypsin or SLIGRL-NH2 resulted in a transient rise of the intracellular Ca2+ level and trypsin-induced morphological changes were blocked by BAPTA, a Ca2+ chelator. In addition, a protein kinase C (PKC) inhibitor, bisindolylmaleimide significantly inhibited the trypsin-induced morphological changes, whereas activation of PKC by phorbol-12-myristate-13-acetate acted as trypsin. Taken together, these results suggest that activation of PAR-2 by trypsin caused reversal of stellation in cultured astrocytes, in part, via the mobilization of intracellular Ca2+ and activation of PKC. PMID- 16256234 TI - Differential expression of proteins in cold-adapted log-phase cultures of Escherichia coli incubated at 8, 6 or 2 degrees C. AB - Temperature is used to control the growth of microorganisms in foods. The minimum temperature for sustained growth of Escherichia coli is 7 degrees C. E. coli cells in the logarithmic phase of growth at 15 degrees C were incubated at 8, 6 or 2 degrees C. The cells grew with the formation of filaments at the two higher temperatures, but did not grow at 2 degrees C. In order to investigate more thoroughly the nature of filament formation in E. coli at temperatures near the minimum temperature for sustained growth, cells were harvested after 1 day at 2 degrees C or at times up to 4 or 8 days at 8 or 6 degrees C, respectively. Proteins extracted from the cells were separated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE), and spots containing differentially expressed proteins were identified by quadropole time-of-flight tandem (Q-ToF-2) mass spectrometry. For most of the identified proteins, the amounts were not substantially different in cells grown at 15 degrees C or incubated at 2 degrees C. In cells incubated at 8 or 6 degrees C, proteins associated with stress responses, the tricarboxylic acid cycle and electron transport were present in substantially greater amounts, and proteins associated with protein synthesis were present in substantially smaller amounts than in cells grown at 15 degrees C. These findings suggest that the stringent response is induced in E. coli incubated at temperatures near the minimum for growth, so the formation of filaments at those temperatures may be a result of the stringent response. PMID- 16256236 TI - Genetic analysis of the function of the plum pox virus CI RNA helicase in virus movement. AB - The CI protein forms the cylindrical inclusions typical of potyviral infections and is involved in genome replication and virus movement. In this work, we have analyzed the effect of a series of point mutations at the N-terminal region of the CI protein of Plum pox virus (PPV) on the enzymatic activities and the self interaction ability of the protein, and on virus replication and movement. DD3,4AA mutation, which had no apparent effects on ATPase and RNA helicase activities in vitro, and on virus replication in protoplasts, drastically impaired cell-to-cell spread of the virus. The effect of KK101,102AA mutation was host-specific. While no signals of virus infection were detected in Chenopodium foetidum inoculated with PPV KK101,102AA, the mutation caused a moderate effect on short distance movement in Nicotiana benthamiana and N. clevelandii, which resulted in a more drastic disturbance of systemic spread. None of the mutations analyzed abolished PPV CI self-interaction in the yeast Two-Hybrid system, but they caused a notable reduction in the binding strength, which appears to positively correlate with their effect on virus movement, suggesting that CI-CI interactions required for RNA replication and virus movement could be rather different. PMID- 16256235 TI - Granule development and performance in sucrose fed anaerobic baffled reactors. AB - Two 90 L anaerobic baffled reactors were used to study the granulation of sludge and the effect of the organic loading rate and NaHCO3/COD ratios on reactor performance. Furthermore, it was determined whether an anaerobic baffled reactor would promote phase separation and if additive of bentonite or granular active carbon was capable of enhancing granule formation. In order to minimize feed variations, and have a totally biodegradable substrate, a synthetic sucrose substrate was used. Granulation was achieved in both reactors within 75 days. However, the granules from the granular active carbon amended reactor appeared earlier and were larger and more compact. The reactors were maintained at a hydraulic retention time of 20 h during performance study stage. The results showed that when organic loading rate were changed from 2.15 to 6.29 kg COD m( 3)day(-1), chemical oxygen demand (COD) removal was not decreased (91-93%), but a slight increase in effluent COD was observed. It was found that the COD removals were generally good (87-92%) and had not obviously change with the decreasing NaHCO3/COD ratios. From the bacterial distribution and the concentration of volatile fatty acids in four compartments, it was concluded that a separation of phases occurred within the anaerobic baffled reactors. PMID- 16256237 TI - AIDS vaccine: Intranasal immunization using inactivated HIV-1-capturing core corona type polymeric nanospheres. AB - Polymeric nanospheres have been widely used in biomedical applications, such as drug, gene and vaccine delivery systems. Nanospheres with entrapped antigens have recently been shown to possess significant potential as vaccine delivery systems and adjuvants. We previously reported that concanavalin A-immobilized polystyrene nanospheres (Con A-NS) could efficiently capture HIV-1 particles and intranasal immunization with inactivated HIV-1-capturing nanospheres (HIV-NS) induced vaginal anti-HIV-1 IgA antibody responses in mice. In addition, vaginal washes from intranasally immunized mice were capable of neutralizing HIV-1. Moreover, simian/human immunodeficiency virus KU-2-capturing nanospheres (SHIV-NS) immunized macaques exhibited partial protection when vaginally and systemically challenged with pathogenic viruses. HIV-NS is suggested to be particularly suitable to enhance antigen delivery to dendritic cells (DCs). In this study, we investigated the mucosal antibody response in mice after the intravaginal or intranasal immunization in detail with using different sized (360, 660, 940 and 1230 nm) HIV-NS. The amount of immobilized Con A to NS was dependent on the surface area of the particle. Moreover, Con A-NS with different sizes could equally capture inactivated HIV-1. Intravaginal or intranasal immunization by HIV NS with diameters ranging 360 to 1230 nm significantly induced vaginal antibody responses. However, significant differences on vaginal anti-HIV-1 gp120 IgA and IgG antibodies were not found after intravaginal or intranasal immunization with different sized HIV-NS. These results suggest that HIV-NS provides an efficient vaccine delivery system for the induction of a mucosal immune response and the development of a mucosal vaccine. PMID- 16256238 TI - Inhibition of metastatic tumor growth by targeted delivery of antioxidant enzymes. AB - To develop effective anti-metastatic therapy, targeted or sustained delivery of catalase was examined in mice. We found that mouse lung with metastatic colonies of adenocarcinoma colon26 cells exhibited reduced catalase activity. The interaction of the tumor cells with macrophages or hepatocytes generated detectable amounts of ROS, and increased the activity of matrix metalloproteinases. Hepatocyte-targeted delivery of catalase was successfully achieved by galactosylation, which was highly effective in inhibiting the hepatic metastasis of colon26 cells. PEGylation, which increased the retention of catalase in the circulation, effectively inhibited the pulmonary metastasis of the cells. To examine which processes in tumor metastasis are inhibited by catalase derivatives, the tissue distribution and proliferation of tumor cells in mice was quantitatively analyzed using firefly luciferase-expressing tumor cells. An injection of PEG-catalase just before the inoculation of melanoma B16-BL6/Luc cells significantly reduced the number of the tumor cells in the lung at 24 h. Daily dosing of PEG-catalase greatly inhibited the proliferation of the tumor cells, and increased the survival rate of the tumor-bearing mice. These results indicate that targeted or sustained delivery of catalase to sites where tumor cells metastasize is a promising approach for inhibiting metastatic tumor growth. PMID- 16256239 TI - Physical activity barriers in diabetes: development and validation of a new scale. AB - To develop and validate a questionnaire measuring perceived Barriers to Physical Activity in Diabetes (type 1) or BAPAD1. Initially, an open-ended questionnaire was filled by 36 patients. The modal accessible beliefs obtained on this pilot study were analysed and a scale composed of 12 items (BAPAD1) was developed and validated. Seventy-four type 1 diabetic patients filled the BAPAD1 scale. Cronbach alpha coefficient was 0.85 and the correlation between the test-retest scores was 0.84, both indicating adequate reliability of the barriers scale. Each item of BAPAD1 scale displayed very good item characteristic curve except for item 12, which was withdrawn. The test reliability curve indicated that the BAPAD1 scale is informative (value>or=0.82) at all levels of perceived barriers toward physical activity. Moreover, among diabetic-related items, the risk of hypoglycemia showed a particularly good item characteristic curve. In summary, the BAPAD1 scale presents excellent psychometric proprieties and among diabetic related items, the risk of hypoglycemia should be considered as a significant target to overcome in order to increase physical activity. This new validated tool should be useful in identifying the most salient barriers toward the practice of physical activity and thus, permit more focused intervention in order to overcome those barriers. PMID- 16256240 TI - Nitric oxide might be involved in the chloroquine-improved insulin sensitivity: old treatment for global danger. PMID- 16256241 TI - Glucose-stimulated insulin response in non-diabetic patients with lipoprotein lipase deficiency and hypertriglyceridemia. AB - Elevations in plasma triglyceride (TG) and free fatty acid (FFA) concentrations are generally thought to play a role in the pathogenesis of insulin-resistant diabetes. The objective of this study was to investigate the relationship between hypertriglyceridemia and glucose-stimulated insulin responsiveness in non diabetic patients. Forty subjects were divided into three BMI-matched groups as follows: one group consisted of 8 patients with a lipoprotein lipase (LPL) deficiency, another consisted of 12 patients with hypertriglyceridemia and a third consisted of 20 subjects with normal TG levels. In response to a 75 g oral glucose tolerance test, plasma insulin levels in the LPL-deficient subjects were higher (106+/-11 microU/ml) than those in the hypertriglyceridemic (69+/-16 microU/ml) and normolipidemic (29+/-3 microU/ml) subjects, at 30 min. On the other hand, their plasma glucose levels (127+/-6 mg/dl) were less than those seen in the normolipidemic group (165+/-9 mg/dl) after 90 min. Thus, LPL-deficient subjects with hypertriglyceridemia displayed an enhanced glucose-stimulated insulin response as well as lower blood glucose levels, the latter of which is not generally seen in those with hypertriglyceridemia and normolipidemia. PMID- 16256243 TI - HIV surveillance in theory and practice: assessing the acceptability of California's non-name HIV surveillance regulations. AB - In 2002, California adopted a non-name system for HIV case reporting. This study focused on the acceptability of a non-name reporting system among key stakeholders implementing the system. We conducted qualitative research during the pre- and post-implementation period of the non-name HIV reporting regulations. During both study periods we conducted key informant in-depth interviews (n = 48 and 52, respectively) with health department surveillance staff, laboratory personnel, health care providers and clinic staff; and we conducted four focus group discussions (n = 28 and 30, respectively) with representatives of community-planning group members and advocacy groups. We found that overall, California's non-name HIV reporting regulations were acceptable to most key stakeholders. Acceptability of a non-name system was highest among advocates and healthcare providers. Views of health department staff varied across the four counties, with some expressing a strong preference of a names based system and others accepting the non-name system. PMID- 16256242 TI - Impact of modified glucose target and exercise interventions on vascular risk factors. AB - Potent glucose-lowering medications other than metformin may impede weight loss in type 2 diabetes (T2D). Supervised exercise programs improve glycemic control without significantly enhancing weight loss; their impact on fitness and blood pressure in T2D remains unclear. In this pilot study, 42 type 2 diabetes patients were randomized to (i) liberalized (10 mmol/l) OR strict (7 mol/l) preprandial glucose thresholds for adjustment of medication other than metformin and (ii) dietary counseling with OR without supervised exercise (2 x 2 factorial design). Weight (-1.2% versus -0.3%, p=0.38) and hemoglobin A1C changes (-0.8% versus 0%, p=0.37) were similar for glucose threshold-defined groups. Dietary counseling with supervised exercise participants had greater improvement in mean arterial pressure than those not randomized to an exercise program (-3.3% versus 1.1%, p=0.02). Overall weight (-1.5% versus 0%, p=0.06) and fitness changes (5.4% versus 1.5%, p=0.18) were not significantly different between these groups, but weight (-1.6% versus 0%, p=0.03) and fitness changes (21.3% versus 1.5%, p=0.03) were significantly greater among those who attended >or=75% of exercise classes. Liberalizing preprandial thresholds neither enhances weight loss nor compromises hemoglobin A1C. T2D patients who consistently participate in supervised exercise programs may experience modest weight loss and significant improvements in fitness and blood pressure. PMID- 16256244 TI - Determining the error of dose estimates and minimum and maximum acceptable concentrations from assays with nonlinear dose-response curves. AB - A method is described here that uses a modified Monte-Carlo method to provide an improved estimate of the confidence bounds of concentration estimates. This method accommodates even strongly nonlinear curve models, such as the five parameter logistic model, in contrast to the common but often poor approach of linearizing the regression problem and using linear theory to obtain the confidence bounds. The method uses an interpolation technique to reduce artifacts in the precision profile due to small simulation sample sizes and proximity to horizontal asymptotes in the curve model. The paper also describes how to define and calculate the minimum and maximum acceptable concentrations of dose-response curves by locating the concentrations where the size of the error, defined in terms of the size of the concentration confidence interval, exceeds the threshold of acceptability determined for the application. PMID- 16256246 TI - Modification of adenosine A1 and A2A receptor density in the hippocampus of streptozotocin-induced diabetic rats. AB - Adenosine A(1) and A(2A) receptors are neuromodulatory systems that can control mnemonic behavior, which is modified by diabetes. Since the density of these adenosine receptors can change upon chronic noxious brain conditions, we now tested if the density of A(1) and A(2A) receptors was modified in the hippocampus of streptozotocin-induced diabetic rats. The binding density of the selective A(1) receptor antagonist, (3)H-DPCPX, was decreased by 36% in total hippocampal membranes 7 days after induction of diabetes and this down-regulation was maintained after 30 and 90 days, which was also confirmed by Western blot analysis of A(1) receptor immunoreactivity. In contrast, the binding density of the selective A(2A) receptor antagonist, (3)H-SCH 58261, was enhanced by 83% in total hippocampal membranes 7 days after induction of diabetes and this up regulation persisted after 30 and 90 days. These results show that the balance between inhibitory A(1) and facilitatory A(2A) adenosine receptors is modified in the hippocampus of streptozotocin-induced diabetic rats. Thus, the most abundant A(1) receptors are down-regulated and there is an up-regulation of A(2A) receptors, suggesting a gain of function of hippocampal A(2A) receptors compared to A(1) receptors in diabetes, as has been observed in other chronic noxious brain conditions where A(2A) receptor blockade affords robust neuroprotection. PMID- 16256245 TI - Selective side-chain modification of cysteine and arginine residues blocks pathogenic activity of HIV-1-Tat functional peptides. AB - Extracellular Tat protein of HIV-1 activates virus replication in HIV-infected cells and induces a variety of host factors in the uninfected cells, some of which play a critical role in the progression of HIV infection. The cysteine-rich and arginine-rich basic domains represent key components of the HIV-Tat protein for pathogenic effects of the full-length Tat protein and, therefore, could be ideal candidates for the development of a therapeutic AIDS vaccine. The present study describes selective modifications of the side-chain functional groups of cysteine and arginine amino acids of these HIV-Tat peptides to minimize the pathogenic effects of these peptides while maintaining natural peptide linkages. Modification of cysteine by introducing either a methyl or t-butyl group in the free sulfhydryl group and replacing the guanidine group with a urea linkage in the side chain of arginine in the cysteine-rich and arginine-rich Tat peptide sequences completely blocked the ability of these peptides to induce HIV replication, chemokine receptor CCR-5 expression, and NF-kappaB activity in monocytes. Such modifications also inhibited angiogenesis and migration of Kaposi's sarcoma cells normally induced by Tat peptides. Such chemical modifications of the cysteine-rich and arginine-rich peptides did not affect their reactivity with antibodies against the full-length Tat protein. With an estimated 40 million HIV-positive individuals worldwide and approximately 4 million new infections emerging every year, a synthetic subunit HIV-Tat vaccine comprised of functionally inactive Tat domains could provide a safe, effective, and economical therapeutic vaccine to reduce the progression of HIV disease. PMID- 16256247 TI - Firing of 5-HT neurones in the dorsal and median raphe nucleus in vitro shows differential alpha1-adrenoceptor and 5-HT1A receptor modulation. AB - The median raphe nucleus and dorsal raphe nucleus together are the major source of ascending 5-HT projections. Here, using in vitro extracellular single unit electrophysiology we examined the responses of individual neurones in the rat median raphe nucleus and dorsal raphe nucleus to alpha(1)-adrenoceptor and 5 HT(1A) receptor activation and made comparisons between the two nuclei. In the presence of the alpha(1)-adrenoceptor agonist phenylephrine (1microM) all spontaneously active neurones recorded in the median and dorsal raphe nuclei fired slowly (<5Hz) and regularly. Most were inhibited by 5-HT (10-50microM), although a few were excited by 5-HT. 5-HT-induced inhibition was attenuated by the 5-HT(1A) receptor antagonist, WAY100635 (100nM). Compared to those in the dorsal raphe nucleus, the neurones in the median raphe nucleus which were inhibited by 5-HT had: (1) lower basal firing rates in the continuous presence of phenylephrine (1microM), (2) smaller excitatory responses to higher concentrations of phenylephrine (3-10microM), (3) smaller excitatory responses to brief application of norepinephrine (10-100microM) and (4) smaller inhibitory responses to 5-HT (10-50microM). The lower sensitivity of median raphe neurones to alpha(1)-adrenoceptor excitation and 5-HT(1A) receptor inhibition will have consequences for 5-HT neurotransmission in forebrain regions innervated by the two nuclei. PMID- 16256248 TI - Dietary intake of unsaturated fatty acids and age-related cognitive decline: a 8.5-year follow-up of the Italian Longitudinal Study on Aging. AB - There is evidence from a population-based study of an inverse relationship between monounsaturated fatty acids (MUFA) energy intake and age-related cognitive decline (ARCD), while high polyunsaturated fatty acids (PUFA) intake was positively associated with cognitive impairment in elderly subjects. We investigated the possible role of MUFA and PUFA on age-related cognitive changes. A population-based, prospective study was carried out on 278, 186, and 95 nondemented elderly subjects (65-84 years) evaluated for global cognitive functions (Mini-Mental State Examination, MMSE) at the first (1992-1993), second (1995-1996), and third survey (2000-2001), respectively, from the randomized cohort of Casamassima, Bari, Italy (n=704), one of the eight centers of the Italian Longitudinal Study on Aging (ILSA). MUFA and PUFA intakes were assessed at baseline with a semi-quantitative food frequency questionnaire. High MUFA and PUFA energy intakes and total energy intake were significantly associated with a better cognitive performance in a 8.5-year follow-up. In this prospective population-based study on older nondemented subjects with a typical Mediterranean diet, high MUFA and PUFA intakes appeared to be protective against ARCD. PMID- 16256249 TI - Synthesis and antifungal potential of Co(II) complexes of 1-(2'-hydroxyphenyl) ethylideneanilines. AB - Synthesis of Co(II) complexes from 1-(2'-hydroxyphenyl)-ethylideneaniline and its N-phenyl derivatives has been described and their structures have been established on the basis of elemental analysis and IR studies. The complexes have been screened for their antifungal potential against Alternaria alternata, Fusarium oxysporum and Myrothecium roridum by spore germination inhibition method and the results are discussed. PMID- 16256250 TI - Effect of zinc and vitamin A supplementation on antibody responses to a pneumococcal conjugate vaccine in HIV-positive injection drug users: a randomized trial. AB - HIV-infected individuals have impaired immune responses to vaccines and high rates of pneumococcal disease. The effect of vitamin A and zinc supplementation on the immunogenicity of a 7-valent pneumococcal CRM-197 conjugate vaccine (PC-7) was evaluated in 118 HIV+ injection drug users. Subjects were randomized to oral 400,000 IU vitamin A, 300 mg zinc, vitamin A + zinc, or placebo, then immunized. Geometric mean titer increased 1.3-3.3-fold for all pneumococcal serotypes. PC-7 elicited an immune response in HIV-infected adults but neither vitamin A nor zinc altered the immunogenicity of the evaluated vaccines. PMID- 16256251 TI - Induction of protective immunity against toxoplasmosis in mice by immunization with Toxoplasma gondii RNA. AB - Toxoplasma gondii enters the mucosal surfaces of the host, and so immunity at these sites is of major interest. Due to the compartmentalization of the immune response, systemic immunization does not induce high levels of immunity at mucosal surfaces. Intranasal immunization has been shown to be very effective in inducing both systemic and mucosal immune responses. Immunization with mRNA can induce both humoral and cell-mediated immune responses, both of which are important in conferring immunity to T. gondii. The efficacy of RNA vaccination by the nasal route with T. gondii RNA was evaluated. We assessed the percentage of cumulative survival after an oral challenge with a lethal dose of T. gondii cysts (40 cysts), and the number of brain cysts following a challenge with a sublethal dose of T. gondii 76 K cysts (15 cysts). Vaccinated mice were found to be significantly better protected than non-immunized mice after a challenge with a lethal dose of cysts; and a challenge with a sublethal dose also resulted in fewer brain cysts than in non-immunized mice. Sera and intestinal secretions of immunized mice recognized T. gondii antigens, suggesting that a specific humoral immune response may occur. Moreover, a specific lymphoproliferative response observed in cervical lymph nodes may confer protection. These preliminary findings suggest that RNA vaccination by a mucosal route could be feasible. PMID- 16256252 TI - Inter-taxa differences in root uptake of 103/106Ru by plants. AB - Ruthenium-106 is of potential radioecological importance but soil-to-plant Transfer Factors for it are available only for few plant species. A Residual Maximum Likelihood (REML) procedure was used to construct a database of relative (103/106)Ru concentrations in 114 species of flowering plants including 106 species from experiments and 12 species from the literature (with 4 species in both). An Analysis of Variance (ANOVA), coded using a recent phylogeny for flowering plants, was used to identify a significant phylogenetic effect on relative mean (103/106)Ru concentrations in flowering plants. There were differences of 2,465-fold in the concentration to which plant species took up (103/106)Ru. Thirty-nine percent of the variance in inter-species differences could be ascribed to the taxonomic level of Order or above. Plants in the Orders Geraniales and Asterales had notably high uptake of (103/106)Ru compared to other plant groups. Plants on the Commelinoid monocot clades, and especially the Poaceae, had notably low uptake of (103/106)Ru. These data demonstrate that plant species are not independent units for (103/106)Ru concentrations but are linked through phylogeny. It is concluded that models of soil-to-plant transfer of (103/106)Ru should assume that; neither soil variables alone affect transfer nor plant species are independent units, and taking account of plant phylogeny might aid predictions of soil-to-plant transfer of (103/106)Ru, especially for species for which Transfer Factors are not available. PMID- 16256253 TI - Effects of sulfur dioxide on apoptosis-related gene expressions in lungs from rats. AB - Sulfur dioxide (SO2) is an air pollutant in densely populated areas as well as in areas polluted by coal-fired power plants, smelters, and sulfuric acid factories. In the present study, male Wistar rats were housed in exposure chambers and treated with 14.00+/-1.01, 28.00+/-1.77, and 56.00+/-3.44 mg/m3 SO2 for 6 h/day for 7 days, while control rats were exposed to filtered air in the same condition. The mRNA and protein levels of three apoptosis-related genes (p53 and bax are promoters of apoptosis, whereas bcl-2 is apoptotic suppressor) were analyzed in lungs using a real-time reverse transcription-polymerase chain reaction (real-time RT-PCR) assay and immunohistochemistry method, and caspase-3 activities were detected. The results showed that mRNA levels of p53 and bax were increased in a dose-dependent manner and at the concentrations of 28.00 and 56.00 mg/m3 SO2 the increases were significant (for p53: 1.23-fold at 28 mg/m3 and 1.39 fold at 56 mg/m3; for bax: 1.77-fold at 28 mg/m3 and 2.26-fold at 56 mg/m3, respectively), while mRNA levels of bcl-2 were decreased significantly (0.78-fold at 28 mg/m3 and 0.73-fold at 56 mg/m3) in lungs of rats exposed to SO2. Dose dependent increase of p53 and bax proteins in the lungs was observed after SO2 inhalation, while decrease of bcl-2 protein levels was obtained using immunohistochemistry method. Caspase-3 activities were increased in lungs of rats after SO2 inhalation. These results lead to a conclusion that SO2 exposure can change the expression of apoptosis-related genes, and it suggests that SO2 can induce apoptosis in lung of rat and may have relations with some apoptosis related diseases. Elucidating the expression patterns of those factors after SO2 inhalation may be critical to our understanding mechanisms of SO2 toxicity and helpful for the therapeutic intervention. PMID- 16256254 TI - Improving REACH. AB - The European Commission has proposed a new system for chemicals control, the REACH system. The objectives of REACH with respect to risk assessment can be summarized in the form of two overarching goals. First, REACH aims at improved knowledge about the properties and the uses of individual chemical substances. Secondly, REACH is intended to increase the speed and efficiency of the risk assessment process and to make producers and importers of chemicals responsible for this process. The implementation of the REACH proposal will lead to increased availability of toxicological information but not to the extent that would be needed to achieve a sound scientific basis for risk assessment of individual substances. In this paper, we propose certain amendments of the current REACH proposal. The purpose of these proposals is to improve priority-setting and testing strategies and to simplify the risk assessment procedures. PMID- 16256255 TI - Interface of physical and emotional stress regulation through the endogenous opioid system and mu-opioid receptors. AB - Unraveling the pathways and neurobiological mechanisms that underlie the regulation of physical and emotional stress responses in humans is of critical importance to understand vulnerability and resiliency factors to the development of a number of complex physical and psychopathological states. Dysregulation of central stress response circuits have been implicated in the establishment of conditions as diverse as persistent pain, mood and personality disorders and substance abuse and dependence. The present review examines the contribution of the endogenous opioid system and mu-opioid receptors to the modulation and adaptation of the organism to challenges, such as sustained pain and negative emotional states, which threaten its internal homeostasis. Data accumulated in animal models, and more recently in humans, point to this neurotransmitter system as a critical modulator of the transition from acute (warning signals) to sustained (stressor) environmental adversity. The existence of pathways and regulatory mechanisms common to the regulation of both physical and emotional states transcend classical categorical disease classifications, and point to the need to utilize dimensional, "symptom"-related approximations to their study. Possible future areas of study at the interface of "mind" (cognitive-emotional) and "body" (physical) functions are delineated in this context. PMID- 16256257 TI - Factors involved in the determination of the neurotransmitter phenotype of developing neurons of the CNS: applications in cell replacement treatment for Parkinson's disease. AB - The developmental stages involved in the conversion of stem cells to fully functional neurons of specific neurotransmitter phenotype are complex and not fully understood. Over the past decade many studies have been published that demonstrate that in vitro manipulation of the epigenetic environment of the stem cells allows experimental control of final neuronal phenotypic choice. This review presents the evidence for the involvement of a number of endogenous neurobiochemicals, which have been reported to potently influence DAergic (and other neurotransmitter) phenotype expression in vitro. They act at different stages on the pathway to neurotransmitter phenotype determination, and in different ways. Many are better known for their involvement in other aspects of development, and in other biochemical roles. Their proper place, and precise roles, in neurotransmitter phenotype determination in vivo will no doubt be determined in the future. Meanwhile, considerable medical benefits are offered from producing large, long-term, viable cryostores of self-regenerating multipotential neural precursor cells (i.e., brain stem cells), which can be used for cell replacement therapies in the treatment of degenerative brain diseases, such as Parkinson's disease. PMID- 16256256 TI - A ninety-day oral toxicity study of a new type of processed gum arabic, from Acacia tree (Acacia senegal) exudates, in F344 rats. AB - This study was designed to evaluate and characterize any subchronic toxicity of a new type of gum arabic (SUPER GUM [Acacia(sen)SUPER GUM]), a naturally processed polysaccharide exudate from gum acacia trees (Acacia senegal), when administered to both sexes of F344 rats at dietary levels of 0 (control), 1.25%, 2.5%, and 5.0% (10 rats/sex/group). During the study, the treatment had no effects on clinical signs, survival, body weights, and food and water consumption, or on findings of urinalysis, ophthalmology, hematology, or blood biochemistry. Gross pathology and histopathology exhibited no differences of toxicological significance between control and treated rats. Increased relative cecum (filled) weights, evident in both sexes of 5.0% group and females of 1.25% and 2.5% groups, were considered to be a physiological adaptation. Thus, the results indicated the toxic level of SUPER GUM to be more than 5.0%, and the no observed adverse effect level (NOAEL) was concluded to be 5.0% (3,117 mg/kg body weights/day for males, and 3,296 mg/kg body weights/day for males) from the present study. PMID- 16256258 TI - Prevalence of clinical vaginal candidiasis in a university hospital and possible risk factors. AB - OBJECTIVE: The aim of this study was to evaluate the prevalence of symptomatic vaginal candidiasis and probable predisposing factors in a university-based hospital. STUDY DESIGN: A total of 576 cases of clinical vaginal candidiasis were enrolled in this survey and wet mount preparations, Gram-stained smears and vaginal cultures were assessed. Possible risk factors, such as pregnancy, diabetes mellitus, contraceptive and antibiotic use were evaluated. RESULTS: Clinical vaginal candidiasis was detected in 12.1% of the cases. Candida albicans was isolated in 80.2% of patients and non-albicans species in 19.8%. Pruritus was the most common symptom (85.9%), followed by vaginal discharge (66.1%), soreness (31.1%) and dyspareunia (5.0%). Reproductive age, pregnancy, diabetes, contraception as well as recent antibiotic use correlated positively with both C. albicans and non-albicans isolates. Soreness and dyspareunia were significantly related to non-albicans species. The overall recurrence rate was 8.5%. Recurrences correlated positively to non-albicans infections. CONCLUSIONS: C. albicans was, by far, the predominant yeast isolate. Non-albicans isolates caused significantly more frequent soreness, dyspareunia and recurrences than C. albicans. Clinical and laboratory findings, together with possible predisposing factors must be taken into consideration in order to achieve appropriate treatment. PMID- 16256259 TI - Cervical fetal fibronectin as a predictor of first trimester pregnancy outcome in unexplained recurrent miscarriage. AB - OBJECTIVE: To determine whether cervical fetal fibronectin is a reliable predictor of first trimester pregnancy outcome in patients with unexplained recurrent miscarriage. STUDY DESIGN: A prospective observational study was carried out on 49 pregnant women with a history of unexplained recurrent miscarriage. In all participants the presence of fetal fibronectin in the cervical secretion was determined with a qualitative rapid immunoassay. The outcome of the first trimester pregnancy was recorded a successful outcome was a pregnancy that progressed beyond 12 weeks of gestation; a miscarriage referred to a pregnancy loss in the first 12 weeks. RESULTS: Of the 49 subjects screened, fetal fibronectin was positive in 17 and negative in 32. Overall, 14 pregnancies resulted in fetal loss before the 12th week of gestation. Fetal cervical fibronectin was positive in 6 of the 14 patients who miscarried and in 11 of the 35 in whom outcome was successful. As predictor of first trimester pregnancy outcome the test had a sensitivity and a specificity of 43% and 69% and positive and negative predictive values of 35%, and 75%, respectively. Subgroup analysis by number of previous miscarriages and maternal age gave similar values. CONCLUSION: This study examines the possible value of cervical fetal fibronectin in predicting first trimester pregnancy outcome. We conclude that the occurrence of positive or negative fetal cervical fibronectin test has only limited predictive value and therefore its use cannot be considered for clinical application. PMID- 16256260 TI - Efficacy and complications in the surgical treatment of stress urinary incontinence by insertion of a silicone-coated polyester tape (Lift). AB - OBJECTIVES: To evaluate the efficacy and safety of a surgical treatment for stress urinary incontinence by implantation of a silicone-coated polyester tape (Lift). MATERIALS AND METHODS: This retrospective study included 72 female patients having had a suburethral silicone-coated polyester tape inserted as treatment for stress urinary incontinence, combined or not with pelvic surgery. We recorded the patient's characteristics, the surgical procedure, the short and long-term results and complications. RESULTS: Seventy-two patients were operated, 60 of whom were fully evaluated. The average follow-up was 17 months. On 48 patients (80%) the treatment was successful, 3 (5%) were improved, and 9 (15%) were regarded as a failure. Dysuria occurred in six (10%) patients, five were de novo, and one was persistent. Ten patients (16.6%) presented de novo urge incontinence. The main complication was a higher rate of severe infections, accompanied by defective healing (4, i.e. 6.7%). CONCLUSION: The procedure using a silicone-coated polyester tape seems to be efficient, but insufficiently secure. This higher rejection rate leads us to prefer other synthetic materials proved to be better tolerated. PMID- 16256261 TI - Opposing effects of NaCl on reversibility and thermal stability of halophilic beta-lactamase from a moderate halophile, Chromohalobacter sp. 560. AB - Beta-lactamase from a moderately halophilic organism is expected to show salt dependent stability. Here we examined the temperature-dependence of stability at different salt concentrations using circular dichroism (CD) and enzyme activity. NaCl showed opposing effects on melting temperature and reversibility of the thermal melting. Increasing NaCl concentration greatly increased the melting temperature from, e.g., 41 degrees C in the absence of NaCl to 61 degrees C in 3 M NaCl. Conversely, reversibility decreased from 92% to 0% in the corresponding NaCl solutions. When beta-lactamase was heated at different temperatures and NaCl concentrations, the activity recovery followed the reversibility, not the melting temperature. Heating beta-lactamase at 63 degrees C, slightly above the onset temperature of melting in 2 M NaCl and far above the melting in 0.2 M NaCl, showed a much greater recovery of activity in 0.2 M NaCl than in 2 M NaCl, again consistent with the reversibility of melting. PMID- 16256263 TI - Assessment of risk to aquatic biota from elevated salinity -- a case study from the Hunter River, Australia. AB - An ecological risk assessment was performed on salinity levels of the Hunter River and its tributaries to respond to concerns that high salinity may be damaging aquatic ecosystems. Probabilistic techniques were used to assess likelihood and consequence, and hence the risk to aquatic biota from salinity. Continuous electrical conductivity distributions were used to describe the likelihood that high salinity would occur (exposure dataset) and toxicity values were compiled from the limited literature sources available to describe the consequence of high salinity (effects dataset). The assessment was preliminary in the sense that it modelled risk on the basis of existing data and did not undertake site-specific toxicity testing. Some sections of the Hunter River catchment have geologies that are saline because of their marine origins. Catchment development has increased the liberation rates of salts into surface waters. Such modifying activities include coal-mining, power generation and land clearing. The aquatic biota of tributaries had a greater risk of impairment from high salinity than that of the Hunter River. High salinities in the tributaries were attributed to the combined factors of naturally saline geologies, increased liberation of salts due to modification of the landscape, and reduced dilution by flushing flows. A salinity guideline trigger value of 1100 mg L(-1) was recommended. PMID- 16256262 TI - Management of mine spoil for crop productivity with lignite fly ash and biological amendments. AB - Long-term field trials using lignite fly ash (LFA) were carried out in rice crops during the period 1996-2000 at Mine I, Neyveli Lignite Corporation, Tamil Nadu. LFA, being alkaline and endowed with an excellent pozzolanic nature, silt loam texture, and plant nutrients, has the potential to improve the texture, fertility, and crop productivity of mine spoil. The rice crops were the first, third, fifth, and sixth crops in rotation. The other crops, such as green gram (second) and sun hemp (fourth), were grown as green manure. For experimental trials, LFA was applied at various dosages (0, 5, 10, 20, 50, 100, and 200 t/ha), with and without press mud (10 t/ha), before cultivation of the first crop. Repeat applications of LFA were made at the same dosages in treatments of up to 50 t/ha (with and without press mud) before cultivation of the third and fifth crops. Press mud, a lightweight organic waste product from the sugar industry, was used as an organic amendment and source of plant nutrients. Also, a recommended dosage of chemical fertilizer, along with gypsum, humic acid, and biofertilizer as supplementing agents, was applied in all the treatments, including control. With one-time and repeat applications of LFA, from 5 to 20 t/ha (with and without press mud), the crop yield (grain and straw) increased significantly (p < 0.05), in the range from 3.0 to 42.0% over the corresponding control. The maximum yield was obtained with repeat applications of 20 t/ha of LFA with press mud in the third crop. The press mud enhanced the yield in the range of 1.5-10.2% with various dosages of LFA. The optimum dosage of LFA was 20 t/ha for both one-time and repeat applications. Repeat applications of LFA at lower dosages of up to 20 t/ha were more effective in increasing the yield than the corresponding one-time applications of up to 20 t/ha and repeat applications at 50 t/ha. One-time and repeat applications of LFA of up to 20 t/ha (with and without press mud), apart from increasing the yield, evinced improvement in the texture and fertility of mine spoil and the nutrient content of crop produce. Furthermore, some increase in the content of trace and heavy metals and the level of gamma-emitters in the mine spoil and crop produce was observed, but well within the permissible limits. The residual effect of LFA on succeeding crops was also encouraging in terms of eco-friendliness. Beyond 20 t/ha of LFA, the crop yield decreased significantly (p < 0.05), as a result of the formation of hardpan in the mine spoil and possibly the higher concentration of soluble salts in the LFA. However, the adverse effects of soluble salts were annulled progressively during the cultivation of succeeding crops. A plausible mechanism for the improved fertility of mine spoil and the carryover or uptake of toxic trace and heavy metals and gamma-emitters in mine spoil and crop produce is also discussed. PMID- 16256264 TI - Integrated treatment and recycling of stormwater: a review of Australian practice. AB - With the use of water approaching, and in some cases exceeding, the limits of sustainability in many locations, there is an increasing recognition of the need to utilise stormwater for non-potable requirements, thus reducing the demand on potable sources. This paper presents a review of Australian stormwater treatment and recycling practices as well as a discussion of key lessons and identified knowledge gaps. Where possible, recommendations for overcoming these knowledge gaps are given. The review of existing stormwater recycling systems focussed primarily on the recycling of general urban runoff (runoff generated from all urban surfaces) for non-potable purposes. Regulations and guidelines specific to stormwater recycling need to be developed to facilitate effective design of such systems, and to minimise risks of failure. There is a clear need for the development of innovative techniques for the collection, treatment and storage of stormwater. Existing stormwater recycling practice is far ahead of research, in that there are no technologies designed specifically for stormwater recycling. Instead, technologies designed for general stormwater pollution control are frequently utilised, which do not guarantee the necessary reliability of treatment. Performance modelling for evaluation purposes also needs further research, so that industry can objectively assess alternative approaches. Just as many aspects of these issues may have impeded adoption of stormwater, another impediment to adoption has been the lack of a practical and widely accepted method for assessing the many financial, social and ecological costs and benefits of stormwater recycling projects against traditional alternatives. Such triple bottom-line assessment methodologies need to be trialled on stormwater recycling projects. If the costs and benefits of recycling systems can be shown to compare favourably with the costs and benefits of conventional practices this will provide an incentive to overcome other obstacles to widespread adoption of stormwater recycling. PMID- 16256265 TI - Investigations of different kinetic models for zinc ions uptake by a natural zeolitic tuff. AB - The kinetics of uptake of zinc ions from aqueous solutions by natural zeolitic tuff has been investigated. Batch experiments at constant temperature and hydrodynamic conditions have been performed. A decrease in the initial zinc concentration in aqueous solutions prolongs the time needed for equilibrium. Various kinetic models including the film-diffusion model, the surface diffusion model and the heterogeneous diffusion model have been tested for the description of the experimental results of zinc concentration in the solution over time. Diffusion through the film and diffusion through the surface of the particle could be the rate limiting steps at the initial reaction time. However, the heterogeneous diffusion model seems to be the best model providing a satisfactory fitting of the experimental results from the beginning of the process to the point of equilibrium, particularly at lower initial zinc concentrations. PMID- 16256266 TI - Sediment-based investigation of naturally or historically eutrophic lakes -- implications for lake management. AB - Implementation of the EU Water Framework Directive will call for new lake monitoring and management strategies. Therefore, different methods need to be tested in order to achieve reliable assessment of lake background conditions and water quality. Sediment-based techniques provide one such tool for lake management. In this work, 10 lakes, presumed to be naturally eutrophic, were investigated with a paleolimnological short core study. The aim of the study was to examine the composition of the diatom assemblages in their natural state, estimate their change over time and assess the background nutrient levels. One sediment profile from each lake was divided into six sub-samples that were analyzed for diatoms (60 samples). Diatom-based inference models were applied to reconstruct the past total phosphorus concentration and assess the eutrophication. The results indicated that all the lakes studied had already been nutrient-rich before the impact of modern agriculture. However, diatom assemblages have changed remarkably over time and total phosphorus concentrations have generally increased, so at present only two of the study lakes are close to their natural status. This suggests that naturally eutrophic lakes will probably require management actions to fulfill the new directive requirements in the future. PMID- 16256267 TI - An anti-tumor role for cGMP-dependent protein kinase. AB - This study compared Type-1 cGMP-dependent protein kinase (PKG) expression in normal and tumor tissues and examined PKG function in tumor growth. Studies with a cDNA array revealed that PKG expression was reduced in many tumors compared to respective normal tissue. This decrease in PKG expression was confirmed using quantitative RT-PCR and western blotting of matched colon specimens from normal epithelium and tumor tissue, and also in colon derived cell lines where luciferase reporter analysis revealed that the decreased expression occurred at the transcriptional level. Using SW620 colon carcinoma cells engineered for inducible expression of PKG1beta, it was found that exogenous PKG1beta lead to decreased tumor growth and invasiveness in nude mouse xenografts. PMID- 16256268 TI - Continuous depolarization induces choline acetyltransferase activity in septal and hippocampal co-cultured embryonic rat neurons. AB - We tested the influence of continuous high-K+ treatment on acetylcholine (ACh) release and choline acetyltransferase (ChAT) activity on septal cell culture, and septal and hippocampal cell co-culture obtained from rat embryos. Continuous 9 mM K+ treatment did not affect ACh release and ChAT activity in septal culture, but increased ACh release in co-culture without affecting ChAT activity. A slight increase in extracellular K+ concentration, therefore, induced neuronal excitation. Continuous 55 mM K+ treatment increased ACh release in septal culture. This effect was due to direct excitation of septal neurons. In co culture, 55 mM K+ treatment increased both ACh release and ChAT activity. These results indicate that hippocampal neurons are indispensable for the depolarization-induced increase in ChAT activity in the early stage of developing septal cholinergic neurons. PMID- 16256269 TI - Blood-brain barrier permeability to the neuroprotectant oxyresveratrol. AB - We investigated to what extent the antioxidative hydroxystilbene oxyresveratrol (trans-2,3',4,5'-tetrahydroxystilbene, OXY), that we showed earlier to be strongly neuroprotective in a stroke model, may cross the blood-brain barrier (BBB) in healthy rats and in subjects submitted to focal infarction. Tissue extraction and in vivo microdialysis in the striatum show that systematically applied OXY is able to penetrate the BBB in control animals, but to a low extent. Microdialysis samples from animals that were subjected to a middle cerebral artery occlusion (MCAO) displayed strongly increased OXY levels (more than six fold) in the infarct region as compared to sham-operated rats. Our data show that OXY may exert direct protective effects in the brain by crossing the BBB and may prove an excellent complementary drug for the treatment of neurodegenerative disorders that causally involve oxidative/nitrosative stress, especially in stroke. PMID- 16256270 TI - Functional hyperemic response in the rat visual cortex under halothane anesthesia. AB - To establish a model for functional hyperemia in the rat visual cortex, cortical blood flow responses to flash stimulation were measured with the laser Doppler flow (LDF) technique at various levels of halothane anesthesia. The concentration dependent effect of halothane on arterial pressure and its consequent effect on the hyperemic response were also investigated. Using a stroboscopic light source, 10 flashes at 1 min intervals were delivered to the left eye of 12 Sprague-Dawley rats. LDF responses were measured bilaterally in the monocular primary visual cortex (V1M) at steady state halothane concentrations between 0.4 and 1.4%. In six rats, methoxamine (MX) was infused to prevent halothane-induced hypotension; the remaining rats did not receive MX. In all rats, LDF response to flash commenced within 1s and peaked at 2.5s in the contralateral V1M, but not in ipsilateral V1M. The maximum LDF response was 25% at 0.5% halothane and 12% at 1.4% halothane. In rats without MX infusion, mean arterial pressure (MAP) fell from 138 to 90 mmHg when halothane increased from 0.4 to 1.4%. MX infusion prevented the hypotension, but did not influence the LDF response, suggesting that the halothane's effect was direct rather than pressure-mediated. We demonstrate for the first time, a robust functional hyperemic response to discrete flash stimuli in the primary visual cortex of halothane-anesthetized albino rats that can be measured with LDF over a wide range of halothane concentrations and is not fully suppressed at surgical levels of halothane anesthesia. PMID- 16256271 TI - Increased severity of chemically induced seizures in mice with partially deleted Vitamin D receptor gene. AB - Vitamin D is a neuroactive steroid hormone with multiple functions in the brain. Numerous clinical and experimental data link various Vitamin D-related dysfunctions to epilepsy. Here, we study the role of Vitamin D receptors (VDRs) in experimental epilepsy in mice. To examine this problem, we assessed the seizure profiles in VDR knockout mice following a systemic injection of pentylenetetrazole (70 mg/kg). Overall, compared to the wild-type (WT) 129S1 mice (n=10 in each group), the VDR knockout group significantly demonstrated shorter latencies to the onset, higher Racine scores and increased mortality rates. Our findings suggest that VDRs modulate seizure susceptibility in mice, and that the Vitamin D/VDR endocrine system may be involved in the pathogenesis of epilepsy. PMID- 16256272 TI - Mutations in GABRA1, GABRA5, GABRG2 and GABRD receptor genes are not a major factor in the pathogenesis of familial focal epilepsy preceded by febrile seizures. AB - GABA(A) receptors mutations have been reported in few epilepsy families with febrile seizures (FS) followed by generalized epilepsy. It is not known if such mutations may underlie FS followed by partial epilepsy, which is a more common type of epilepsy. We searched for disease-causing mutations in the genes of the alpha1, alpha5, gamma2 and delta subunits of the GABA-A receptor that were previously shown to contain epilepsy-causing mutations or epilepsy susceptibility polymorphisms. All coding and untranslated exons of these four GABA(A) subunit genes were screened in 74 unrelated patients with familial partial epilepsy preceded by FS. Most patients had temporal lobe epilepsy (TLE). We did not detect any disease-causing mutations that would be consistent with missense, nonsense or splice site mutations in any of the four analyzed genes. We conclude that these genes are not a major genetic factor in familial TLE preceded by FS. PMID- 16256273 TI - Evidence of a role for descending serotonergic facilitation in a rat model of cancer-induced bone pain. AB - Descending modulation of spinal processing plays an important role in chronic pain states. Monoamine pathways comprise a major component of descending controls from the brainstem to the spinal cord. Recent emphasis has been on facilitatory actions mediated by the 5-HT3 receptor. We investigated the effects of spinally administered ondansetron, a selective 5-HT3 receptor antagonist, on electrical- and natural-evoked dorsal horn (DH) neuronal responses in a rat model of cancer induced bone pain (CIBP). Injection of MRMT-1 cells into the tibiae of Sprague Dawley rats was used to model CIBP, whilst sham-operated rats were injected with the cell medium alone. Behavioural testing at regular intervals monitored the development of mechanical allodynia, cold allodynia, and ambulatory-evoked pain. In vivo electrophysiology experiments were carried out 15-17 days after surgery, when there were significant behavioural and neuronal alterations in the cancer animals. Spinally administered ondansetron (10, 50, and 100 microg) had no effect on electrical-evoked neuronal responses, but significantly reduced mechanical- and thermal-evoked responses in both the groups of animals. Furthermore, the effects of ondansetron were significantly greater in cancer animals compared to shams. These results therefore suggest a role for descending serotonergic facilitation in CIBP. PMID- 16256275 TI - Radiotherapy plus adjuvant goserelin improves survival in men with poor prognosis prostate cancer. PMID- 16256274 TI - The efficacy of an oral treatment with paromomycin against an experimental infection with Giardia in calves. AB - A controlled and blinded study was conducted to evaluate the efficacy and safety of a treatment with paromomycin sulphate against an experimental Giardia infection in calves. Animals were infected with 10(5)Giardia cysts of cattle origin and were either treated 11 days later with 25, 50 or 75 mg paromomycin/(kg body weight per day) during 5 consecutive days or not treated (control group). Efficacy was evaluated based on reduction in cyst excretion. Furthermore weight gain and diarrhea scores were monitored. In the group treated with 75 mg/kg per day there was a 100% reduction in cyst excretion until 9 days after the start of the treatment (D9) and a very high reduction (> or =98%) until D13. There was a high reduction (> or =93%) until D9 and D13 in the groups treated with 25 and 50 mg/kg, respectively. The cumulative cyst excretion on D13 was significantly (P<0.05) lower in the groups treated with 75 and 50 mg/kg compared to the control group. Although there was a trend towards higher weight gain and less diarrhea in the treated groups, differences between groups were not significant. No adverse reactions to the paromomycin treatment were recorded. Furthermore, the need for reliable parameters for evaluation of treatments against protozoal infections is emphasised. PMID- 16256276 TI - The potential impact of a low-nitrosamine smokeless tobacco product on cigarette smoking in the United States: estimates of a panel of experts. AB - OBJECTIVE: To predict the impact on tobacco use in the US of a "harm reduction" policy that requires that the smokeless tobacco product meet low nitrosamine standards, but could be marketed with a warning label consistent with the evidence of relative health risks. METHODS: Low nitrosamine smokeless tobacco (LN SLT) and cigarette use are predicted by a panel of experts using a modified Delphi approach. We specify a thought experiment to isolate the changes that would occur after the new LN-SLT policy was implemented. RESULTS: The panel predicted that the new policy would accelerate a decrease in smoking prevalence from 1.3 to 3.1 percentage points over 5 years compared to the current SLT product policy, with greater effects on males than females. Introduction of the new product was also predicted to result in modest additional use of SLT overall, with the greatest increases among males who initiated tobacco use under the new policy. CONCLUSION: An overall consensus was reached that the introduction of a new LN-SLT product under strict regulations would increase SLT use, but reduce overall smoking prevalence. This reduction would likely yield substantial health benefits, but uncertainties surround the role of marketing and other tobacco control policies. PMID- 16256277 TI - Repeat CT imaging and replanning during the course of IMRT for head-and-neck cancer. AB - PURPOSE: Many patients with head-and-neck (H&N) cancer have tumor shrinkage and/or weight loss during the course of radiotherapy. We conducted this retrospective study to determine the dosimetric effects of repeat computed tomography (CT) imaging and replanning during the course of intensity-modulated radiotherapy (IMRT) on both normal tissues and target volumes. METHODS AND MATERIALS: A retrospective chart review identified 13 patients with H&N cancer treated with IMRT who had repeat CT imaging and replanning during the course of radiotherapy. The first IMRT plan for each patient was generated based on the original planning CT scan acquired before the start of treatment. Because of tumor shrinkage or weight loss during radiotherapy, a second CT scan was acquired, and a new plan was generated and used to complete the course of IMRT. CT-CT fusion was used to correct patient positioning differences between the scans. By using a commercial inverse IMRT planning system, a hybrid IMRT plan was generated for each patient by applying the beam configurations of the first IMRT plan (including the intensity profile of each beam) to the anatomy of the second CT scan. The dose-volume histograms of the actual and hybrid IMRT plans were compared using analysis of variance methods for repeated measures. RESULTS: All patients had locally advanced, nonmetastatic Stage III or IV disease, including 6 nasopharynx, 6 oropharynx, and 1 unknown primary site. All patients were treated with concurrent platinum-based chemotherapy. When replanning vs. not replanning was compared, the hybrid IMRT plans (without replanning) demonstrated reduced doses to target volumes and increased doses to critical structures. The doses to 95% (D95) of the planning target volumes of the gross tumor volume (PTVGTV) and the clinical target volume (PTVCTV) were reduced in 92% of patients, by 0.8-6.3 Gy (p=0.02) and 0.2-7.4 Gy (p=0.003), respectively. The maximum dose (Dmax) to the spinal cord increased in all patients (range, 0.2-15.4 Gy; p=0.003) and the brainstem Dmax increased in 85% of patients without replanning (range, 0.6-8.1 Gy; p=0.007). CONCLUSIONS: Repeat CT imaging and replanning during the course of IMRT for selected patients with H&N cancer is essential to identify dosimetric changes and to ensure adequate doses to target volumes and safe doses to normal tissues. Future prospective studies with larger sample sizes will help to determine criteria for repeat CT imaging and IMRT replanning for H&N cancer patients undergoing radiotherapy. PMID- 16256279 TI - Statistical properties of the methods for detecting positively selected amino acid sites. AB - Parsimony and Bayesian methods have been developed for detecting positively selected amino acid sites. It has been reported that the parsimony method is generally conservative. In contrast, the Bayesian method is known to identify more positively selected sites than the parsimony method, especially when the number of sequences analyzed is small, although the interpretation of results obtained from the former method is controversial. Here I show that the likelihood ratio test (LRT) of the Bayesian method corresponds to the parsimony method with window analysis, by analyzing the nucleotide sequences encoding the trans activator (tax) gene of human T-cell lymphotropic virus type I (HTLV-I). It is also indicated that in the parsimony method, the test of selective neutrality using the binomial probability tends to be conservative, but the Monte Carlo simulation is useful for solving this problem. In addition, in the Bayesian method, the bootstrap method appears to produce similar results to the LRT. This information may be useful for improving the methods for detecting positively selected amino acid sites. PMID- 16256278 TI - The use of the mini-mental state examination in recruitment for substance abuse research studies. AB - BACKGROUND: Substance abuse is associated with cognitive impairment. Participation in clinical addiction research can be cognitively demanding. Screening tools can identify cognitively impaired subjects. We examined the use of the mini-mental state examination (MMSE) as an entry criterion in three randomized controlled substance abuse clinical trials. METHODS: In each of the three studies, we calculated the proportion of subjects excluded due to MMSE scores (<21) suggestive of cognitive impairment. We estimated the potential impact on enrollment based on the number of excluded subjects. Separately, for two of the studies, we assessed the impact of cognitive function on participation in follow-up using multivariable logistic regression. RESULTS: Of all persons screened for enrollment, 1.6% (171/10,791) were ineligible based solely on a MMSE score of <21. We estimate that 119 of these 171 ineligible persons would have consented and enrolled. These 119 persons would have represented 9.3% of all enrolled subjects across these studies. For subjects in a study in an inpatient detoxification unit, a higher MMSE score was associated with higher odds (adjusted odds ratio 1.15, 95% CI 1.03-1.30) of completing at least one follow-up assessment. A similar impact on subject follow-up was not observed in a study of medical inpatients with unhealthy alcohol use (adjusted odds ratio 1.01, 95% CI 0.86-1.20). CONCLUSION: Screening for cognitive impairment using the MMSE excludes a small, but substantial, number of persons from addiction research studies. Cognitive ability, as captured by the MMSE may impact follow-up. These data support cognitive screening of substance abuse research subjects. PMID- 16256280 TI - Movement analysis in neonates with spina bifida aperta. AB - INTRODUCTION: In neonates with spina bifida aperta (SBA), leg movements by myotomes caudal to the meningomyelocele (MMC) are transiently observed. It is unclear whether these leg movements relate to functional neural conduction through the MMC. For optimal therapeutical intervention, pathophysiological insight in these transient leg movements seems relevant. If leg movements by myotomes caudal to the MMC concur with the execution of general movements (GMs), functional neural conduction through the MMC is implicated. OBJECTIVE: In neonates with SBA, we aimed to determine whether the transiently present leg movements caudal to the MMC indicate functional neural conduction through the MMC. METHODS: During the perinatal period, fetuses and neonates with SBA (n = 7 and n = 13, respectively) were longitudinally analysed for concurrency between leg movements caudal to the MMC and GMs. To address the integrity of the reflex arc in spinal segments (at, or) caudal to the MMC, tendon leg reflexes were assessed during the first postnatal week. RESULTS: At postnatal day 1, leg movements caudal to the MMC concurred with GMs in 12 of 13 infants. Isolated leg movements were observed in only 3 of these 12 infants (isolated vs. concurrent; p < 0.005). Leg movements concurring with GMs lasted longer than isolated leg movements (median duration = 11 s vs. 2 s; p < 0.05). Between days 1 and 7, tendon leg reflexes (at, or) caudal to the MMC had disappeared in all but 1 neonate. However, leg movements caudal to the MMC remained concurrently present with GMs in all five neonates available for follow-up after day 7. Comparing these leg movements between days 1 and 7 indicated a decreased duration (-44%, p < 0.05). CONCLUSIONS: In neonates with SBA, leg movements caudal to the MMC concur with GMs, indicative of functional neural conduction through the MMC. The disappearance of these leg movements is caused by lower motor neuron dysfunction at the reflex arc, whereas neural conduction through the MMC is still functional. PMID- 16256282 TI - Effect of time of artificial insemination on embryo sex ratio in dairy cattle. AB - The objective of the present study was to examine whether different intervals between insemination and ovulation have an influence on the sex of seven-day-old embryos in dairy cattle. Cows were inseminated once with semen of one of two bulls of proven fertility between 36 h before ovulation and 12 h after ovulation. Time of ovulation was assessed by ultrasound at 4-h intervals. In total, 64 embryos were determined to be male or female. Of these 64 embryos, 51.6% were female. The sex ratio in the various insemination-ovulation intervals (early: between 36 and 20 h before ovulation; intermediate: between 20 and 8 h before ovulation; late: between 8 h before and 12 h after ovulation) did not significantly differ from the expected 1:1 sex ratio (50, 50 and 55% females, respectively). Bull (Bull A and B) and Parity (primiparous and multiparous) had no influence on the expected 1:1 sex ratio either. The number of cell cycles was similar for male and female (P = 0.23) embryos when quality of the embryo (P < 0.0001) was included in the model. The results of this study indicate that, in cattle, the interval between insemination and ovulation does not influence the sex ratio of seven-day-old embryos. PMID- 16256281 TI - Impact of Cyp1a2 or Ahr gene knockout in mice: implications for biomonitoring studies. AB - Studies of the impact of phase 1 enzyme polymorphisms on genetic damage have yielded mixed results. We studied how genetic damage would be altered when specific genes were ablated under low dose conditions. METHODS: Knockouts (KO) were generated from c57bl6/J mice with mutations in Cyp1a2 or Ahr receptor that eliminated gene product function. Animals were treated topically with either 4 aminobiphenyl (4ABP) 10mg/kg, benzo(a)pyrene (BaP) 33.3mg/kg or dibenzo(c,g)carbazole (DBC) 8 mg/kg, and sacrificed after 24h. DNA from livers, skin and/or urinary bladders were isolated and (32)P-post labelled. RESULTS: Cyp1a2-/- mice did not differ in 4ABP DNA adduct levels in either urinary bladder or liver compared to wildtype. There was a sex difference in the organ affected. Cyp1a2 knockout reduced skin BAP adduct levels 50% and AHR knockout reduced skin BAP adduct levels by 90%. There was no impact of either knockout on the levels of DBC-DNA adducts in any tissue. CONCLUSIONS: Ablation of specific metabolizing enzymes had compound- and tissue-specific effects in mice. Phenotypic variability in single CYP enzymes may have minor impact in humans at low doses, but variation in the ability to induce the family of CYPs may have a greater impact. PMID- 16256283 TI - Auditory gap detection in the early blind. AB - For blind individuals, audition provides critical information for interacting with the environment. Individuals blinded early in life (EB) typically show enhanced auditory abilities relative to sighted controls as measured by tasks requiring complex discrimination, attention and memory. In contrast, few deficits have been reported on tasks involving auditory sensory thresholds (e.g., Yates, J.T., Johnson, R.M., Starz, W.J., 1972. Loudness perception of the blind. Audiology 11(5), 368-376; Starlinger, I., Niemeyer, W., 1981. Do the blind hear better? Investigations on auditory processing in congenital or early acquired blindness. I. Peripheral functions. Audiology 20(6), 503-509). A study of gap detection stands at odds with this distinction [Muchnik, C., Efrati, M., Nemeth, E., Malin, M., Hildesheimer, M., 1991. Central auditory skills in blind and sighted subjects. Scand. Audiol. 20(1), 19-23]. In the current investigation we re-examined gap detection abilities in the EB using a single-interval, yes/no method. A group of younger sighted control individuals (SCy) was included in the analysis in addition to EB and sighted age matched control individuals (SCm) in order to examine the effect of age on gap detection performance. Estimates of gap detection thresholds for EB subjects were nearly identical to SCm subjects and slightly poorer relative to the SCy subjects. These results suggest some limits on the extent of auditory temporal advantages in the EB. PMID- 16256284 TI - In vivo demonstration of the absorptive function of the middle ear epithelium. AB - The present study investigated in vivo fluid and ion transport across the middle ear epithelium. The tympanic membrane of rats was punctured under general anesthesia. A capillary tube was fitted to the external auditory canal and the bulla filled with various solutions. Middle ear (ME) fluid volume variations were then measured at constant pressure. When saline was used, a linear decrease of fluid volume was apparent. Replacement of sodium with a non-permeable cation (N methyl-D-glucamin) reduced the absorption rate from 0.065+/-0.008 to 0.019+/ 0.003 microl/min (P<0.05, n=6). Similarly, amiloride (10(-3)M), a sodium channel antagonist, reduced the absorption rate to 0.027+/-0.006 microl/min (P<0.05, n=6). Net absorption was abolished when chloride was substituted with gluconate: 0.008+/-0.004 microl/min (P<0.02, n=6), which might have been related (i) to the role of chloride as a diffusible anion through the paracellular pathway, or (ii) to the secretion of chloride through apical channels. However in this condition, 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid, a chloride channel blocker, did not affect the rate of fluid exchange -0.008+/-0.007 microl/min (P=0.75, n=6). This model provides the first in vivo evidence for the absorptive function of the ME. Fluid introduced into the ME cavity disappears due to active transport through the mucosa. This process is sodium-dependent and can be hindered by high concentration of amiloride. The rate of absorption is high enough to allow total clearance of fluid from the cavity of the middle ear within 13 h. This process might play a role in the maintaining a fluid-free and gas-filled middle ear cavity. PMID- 16256285 TI - [WITHDRAWN: Vaccination against human papillomavirus: a new vaccination for the female adolescent?] AB - Ahead of Print article withdrawn by publisher. PMID- 16256286 TI - Applications of mass spectrometry in early stages of target based drug discovery. AB - Mass spectrometry (MS) has been applied to drug discovery for many years. With the advent of new ionization techniques, MS has emerged as an important analytical tool in identification and characterization of protein targets, structure elucidation of synthetic compounds, and early drug metabolism and pharmacokinetics studies. Two MS-based strategies, function-based and affinity based, have been employed in recent years for screening and evaluation of compounds. In the function-based approach, the effects of compounds on the biological activity of a target molecule are measured. In the affinity-based approach, compounds are screened based on their binding affinities to target molecules. The interaction between targets and compounds can be directly evaluated by monitoring the formation of non-covalent target-ligand complexes (direct detection) or indirectly evaluated by detecting the compounds after separating bound compounds from unbound (indirect detection). Various techniques including high performance liquid chromatography (HPLC)-MS, size exclusion chromatography (SEC)-MS, frontal affinity chromatography (FAC)-MS and desorption/ionization on silicon (DIOS)-MS can be applied. The recent advances, relative advantages, and limitations of each MS-based method as a tool in compound screening and compound evaluation in the early stages of drug discovery are discussed in this review. PMID- 16256287 TI - Physico-chemical characterisation and intrinsic dissolution studies of a new hydrate form of diclofenac sodium: comparison with anhydrous form. AB - Diclofenac sodium is a non-steroidal anti-inflammatory drug widely used in painful and inflammatory diseases. In standard conditions, by exposure to relative humidity even below 60% at 25 degrees C, the anhydrous form DS gives rise to a hydrate species DSH, a tetrahydrate form different from that obtained by crystallisation from water and previously described. The method of preparation and the physico-chemical properties of the hydrate form were investigated. Data from FTIR spectroscopy, X-ray powder diffraction and thermal analysis were used for the identification and the characterisation of DSH. DS and DSH were easily differentiated by their IR spectra, X-ray patterns and thermal behaviour. DSH stability was followed at room temperature over a period of 1 year and under different conditions of temperature to verify the tendency to solid-solid transition and to study its existence range. Solubility and intrinsic dissolution studies were performed to compare the physico-chemical properties of DS and DSH. Differences in solubility and intrinsic dissolution rates were pointed out: these studies showed that DS dissolved faster than DSH. Storage under uncontrolled environmental conditions or contact with water vapour during manufacturing process could thus influence the performance of the final dosage form. PMID- 16256288 TI - Electrochemical study on the behavior of Morin and its interaction with DNA. AB - Voltammetric behavior of Morin was studied in 0.1M HAc-NaAc+50mM KCl (pH 3.4) solution at glassy carbon electrode (GCE) using cyclic voltammetry (CV). Morin showed an irreversible anodic peak at 0.720 V in CV which was involving two electrons and two protons. Also, the interaction of Morin with double-stranded calf thymus DNA (ctDNA) was studied by CV at GCE with an irreversible electrochemical equation. As a result of reaction with ctDNA, the voltammetric peak of Morin was a position shift and the peak current decreased. The diffusion coefficients of both free and binding Morin (D(f)=1.1,086 x 10(-7)cm(2)s(-1) and D(b)=8.2,544 x 10(-9)cm(2)s(-1)), binding constant (K=1.7,765 x 10(7)cm(3)mol( 1)), and binding site size (s=0.8,510) of the Morin-DNA complex were obtained simultaneously by non-linear fit analysis. The results demonstrate that Morin can bind to ctDNA in 0.1M HAc-NaAc+50mM KCl (pH 3.4) solution and the ring B of Morin intercalates between the DNA base pairs. PMID- 16256289 TI - Application of 2-acetylbutyrolactone to spectrofluorimetry: fluorescence properties of Schiff bases derived from 2-acetylbutyrolactone and spectrofluorimetric determination of primary amine-containing compounds. AB - 2-Acetylbutyrolactone (ABL) has been characterized for use as a fluorogenic reagent for the spectrofluorimetric determination of primary amines. The reagent forms strongly fluorescent Schiff bases upon the reaction with primary amines in acid-catalized aqueous solutions or in dimethylformamide (DMF). Sulfamethoxazole (SMX) and ampicillin sodium (AMP Na) were used as model amines of type ArNH(2) and RNH(2), respectively. The reaction conditions, fluorescence spectral properties and the stability of the derivatives have been investigated. The chemistry and the pathway of the reaction have been discussed. Calibration data, accuracy, precision, limits of detection, limits of quantification and other aspects of analytical merit were presented in the text. The utility of ABL for the analysis of the model drugs in pharmaceutical preparations was demonstrated. The results indicated that the proposed methods are equally accurate and precise as the official or other reported methods. PMID- 16256290 TI - Changes in personal control as a predictor of quality of life after pulmonary rehabilitation. AB - OBJECTIVE: Perceptions of mastery and self-efficacy may be related to better outcomes in pulmonary rehabilitation in patients with chronic obstructive pulmonary disease (COPD). This study examined (1) whether patients with COPD improved during a rehabilitation programme with respect to quality of life (QoL) and perceptions of self-efficacy and mastery, and (2) whether increased perceptions of mastery and self-efficacy contributed to a higher QoL after rehabilitation. METHODS: Thirty-nine consecutive COPD patients (aged 60.5 +/- 9.0) were included from a rehabilitation centre and completed self-report questionnaires assessing symptoms, QoL, and perceptions of personal control. RESULTS: COPD patients improved during rehabilitation in overall QoL and self efficacy, although no significant changes were found in QoL domains and mastery. Changes in self-efficacy during rehabilitation contributed to the explanation of the social and psychological functioning QoL domains. CONCLUSION: Even in seriously impaired COPD patients in advanced stages of illness, positive changes in self-efficacy and overall well-being can be established during rehabilitation. Changes in self-efficacy were related to a better QoL, suggesting the importance of personal control in the adjustment to COPD. PRACTICE IMPLICATIONS: Focussing more explicitly on the enhancement of perceptions of personal control in COPD patients may be an important aim of pulmonary rehabilitation. PMID- 16256291 TI - Seizing the 9-month moment: addressing behavioral risks in prenatal patients. AB - OBJECTIVE: Our qualitative study explored prenatal care providers' methods for identifying and counseling pregnant women to reduce or stop smoking, alcohol use, illicit drug use, and the risk of domestic violence. METHODS: We conducted six focus groups (five with OB/Gyn physicians, one with nurse practitioners and certified nurse midwives), total N=49, using open-ended questions. Investigators analyzed transcripts to identify and describe themes. RESULTS: Three major themes emerged: (1) specific risk-prevention tactics or strategies exist that are useful during pregnancy; (2) some providers address patients' isolation or depression; and (3) providers can adopt a policy of "just chipping away" at risks. Specific tactics included normalizing risk prevention, using specific assessment techniques and counseling strategies, employing a patient-centered style of smoking reduction, and involving the family. CONCLUSIONS: Providers generally agreed that addressing behavioral risks in pregnant patients is challenging. Patient-centered techniques and awareness of patients' social contexts help patients disclose and discuss risks. PRACTICE IMPLICATIONS: Brief but routine assessment and risk reduction messages require little time of the provider, but can make a big difference to the patient, who may make changes later. PMID- 16256292 TI - Exploring beliefs about prostate cancer and early detection in men and women of three ethnic groups. AB - OBJECTIVES: Practice guidelines support informed or shared decision-making about prostate cancer screening. To compare beliefs across three racial/ethnic categories concerning prostate cancer etiology and risk, screening routines, and shared decision-making, we conducted 12 focus groups. METHODS: Participants were recruited in primary care settings and included 33 African Americans, 35 Hispanics, and 22 non-Hispanic Whites. Of the 90 participants, 53% were male. RESULTS: Groups identified heredity, age, race, sexual activity, and other lifestyle influences as risk factors. Few were aware that prostate cancer is asymptomatic in early stages. Confidence in knowledge of screening routines was high, but included misconceptions supporting initiation of screening at earlier ages and at shorter intervals than professional recommendations. Females encouraged screening of male relatives to protect their health. DISCUSSION AND CONCLUSION: While racial/ethnic groups had similar views and knowledge about screening, African Americans wanted to organize to address the threat of prostate cancer in their communities. Hispanics had awakening awareness of the health risks of prostate cancer. Non-Hispanic Whites were aware of the health threat of prostate cancer, but their approach to health protection was more individual and less community focused than that of African Americans. Participants were not aware of controversy about screening. PRACTICE IMPLICATIONS: Developers of educational materials to support informed or shared decision-making should be aware that initial views of prostate cancer screening are positive. PMID- 16256293 TI - Pediatrician attitudes to exclusion of ill children from child-care centers in Israel: pressure on ambulatory practices. AB - BACKGROUND: The exclusion of ill children from child-care centers may be associated with high social, economic and medical costs. OBJECTIVE: To assess the opinions of pediatricians working in an outpatient setting in Israel on the exclusion/return of children in child-care centers. METHODS: A questionnaire on practices of exclusion/return of children in child-care centers, in general and according to specific signs and symptoms, was administered to a random computer selected cross-sectional sample of 192 primary care community pediatricians in Israel. RESULTS: One hundred and seventy-three pediatricians completed the questionnaires, for a response rate of 90%; 147 were board-certified and 26 were not. About half the pediatricians felt pressured by parents requesting antibiotic therapy to accelerate the return of their sick child to the child-care center. The majority also believed their practice was overloaded by often unnecessary demands for medical notes by the child-care centers before children could return. More than half based their decision to exclude children on "common sense" and the remainder, on accepted guidelines. Except for scabies and lice, there were no significant correlations between the physicians' stipulation for a note on return of the child and the specific illness guidelines. CONCLUSIONS: This study shows that a high proportion of pediatricians based their exclusion practices on "common sense" and personal understanding instead of established guidelines, and that the guidelines did not affect their opinion on the duration of illness that warrant a note. Furthermore, half were subjected to parental pressure to employ inappropriate practices. These findings, combined with our earlier survey of child-care centers staff, indicate that better education of parents and day-care staff about ill child-care-center-exclusion policy in Israel would increase their common understanding with pediatricians. PMID- 16256294 TI - Long term follow-up of c-myc, p53 and proliferation measurements in malignant melanoma. AB - AIMS: We report a prospective study examining the prognostic significance of the c-myc oncoprotein, p53 tumour suppressor gene and proliferation rate measurements in malignant melanoma. METHODS: Flow cytometry (FCM) was used to measure the expression of c-myc, p53 and proliferation parameters in patients who had received an injection of the thymidine analogue bromodeoxyuridine prior to surgery. RESULTS: Sixty-seven patients had successful FCM measurements of the three parameters. c-myc was detected in 97% of patients with a median cell positivity of 62%. The median p53 positivity was 13%. The median potential doubling time (T(pot)) of the tumours wasf 9.4 days. In univariate analysis, each of the parameters showed an association with survival in metatstatic disease with rapid proliferation (p=0.006) or overexpression of c-myc (p=0.038) related to poor survival whereas increased positivity for p53 predicted better survival (p=0.013). CONCLUSIONS: These data indicate that laser cytometric technology can be used to obtain quantitative data on oncoproteins expression and cell proliferation rates in clinical samples of malignant melanoma. PMID- 16256296 TI - [Tumescent local anaesthesia for mastectomy: lidocaine plasma concentration]. PMID- 16256295 TI - [Pregnant woman anaesthetic management in gynaecologic laparoscopic surgery at the maternity hospital of Clermont-Ferrand]. AB - OBJECTIVE: To evaluate the anaesthetic management intended for pregnant women in the field of non obstetric and gynaecologic laparoscopic surgery. STUDY DESIGN: Retrospective and monocentric investigation. PATIENTS AND METHODS: Analysis of the anaesthetic and obstetric files from 27 pregnant women operated on in the establishment, between January 2001 and July 2004. RESULTS: 27 female patients involved in the study. The mean pregnancy term was 15 weeks when laparoscopic surgery was performed, though a single patient was at 30 weeks. The average duration of the surgery was 61 minutes, of which 28 minutes were dedicated to pneumoperitoneum. Twenty-four patients underwent general anaesthesia, the three others underwent locoregional anaesthesia. During the perioperative period no surgical, anaesthetic or obstetric complications were observed. CONCLUSION: Laparoscopic surgery during pregnancy requires double skilled management, both in anaesthesiology and obstetrics. On haemodynamics and breathing, pneumoperitoneum does not induce any additional effects when compared to operations without pregnancy. Except with delivery cases, anaesthetic support in laparoscopic surgery intended for pregnant women eventually does not generate any specific problems, but requires the same rigorous management as the one usually following surgery for pregnant patients. PMID- 16256297 TI - [Postoperative vision loss]. PMID- 16256298 TI - [rFVIIa administration in patient with a left ventricular assistance patient]. AB - We report the case of a left ventricular mechanical assistance (Incor Berlinheart) in a woman that experienced a postoperative non-surgical haemorrhagic complication following a reconstructive pedicled omentoplasty for implanted materials infection. After massive substitutive therapy failure at reducing blood loss and because of an hypovolaemic shock, high dosage (70 microg/kg twice) of recombinant activated factor VII (r-VIIa) was administrated resulting in spectacular cessation of bleeding and haemodynamic restoration. Continuous repeated clinical evaluation, cardiac echography and electronic monitoring of the axial pump device characteristic did not reveal any thromboembolic accident. This observation brings some indirect arguments for safe rFVIIa treatment in this type of pathology with a high thromboembolic risk. PMID- 16256299 TI - [A case of type A botulism]. AB - The neurotoxin produced by Clostridium botulinum, is responsible for botulism. The clinical signs are digestive disorders, pupillar alterations, and peripheral muscular weakness. The failure of thoracic muscles is responsible for the severity of botulism. We describe a case of a 74 year old woman who presented a severe form of botulism, requiring a prolonged intensive care unit stay. PMID- 16256300 TI - [Blocks of the foot]. PMID- 16256301 TI - Quantification of absolute peripheral white blood cells and their subsets in patients with lupus erythematosus: comparison with other inflammatory diseases with and without autoimmune background. AB - To test the value of decreased peripheral leuko-/lymphocytes as screening test for the diagnosis lupus erythematosus (LE). Laboratory routine analyses, and flow cytometry (CD3, CD4, CD8) have been performed in 124 LE-patients. Other subjects included 57 healthy controls, 130 patients with non-autoimmune inflammatory diseases (dermatitis, psoriasis), and 17 patients with another autoimmune disease (progressive systemic sclerosis [PSS]). Numbers of peripheral blood leukocytes (P<0.0005), lymphocytes (P<0.00002), CD3+, and CD3+ CD4+ cells from LE-patients were significantly lower than from controls, patients either with dermatitis, psoriasis or PSS. CD3+ CD8+ cells were significantly diminished in patients with LE and PSS. Decreased numbers of white blood cells, lymphocytes and some of their subsets seem to indicate LE. Patients with PSS only had decreased T-cytotoxic cells. Inflammatory dermatoses lacking the autoimmunity as background have normal cell counts. For screening purpose absolute values of the mentioned blood cell subsets seem to be useful to distinguish both autoimmunity from non-autoimmune inflammatory diseases, and LE from PSS. PMID- 16256302 TI - Serum free cell culture: the free access online database. AB - The cultivation of cells in vitro is an important tool for biomedical research and production purposes. The supplementation of animal/human cell culture media with sera (components) of animal origin remains still standard, providing for e.g. necessary nutrition, shear protection, growth factors and cytokines. Because of undefined composition, risk of contaminations, the cost factor and also animal welfare considerations concerning the production of sera, the conversion to serum free alternatives is promoted by regulatory authorities, industry and the research community in general. To support this trend and to help save one of the scientists most valuable resources -- time -- a data bank was compiled of commercially available formulations, searchable for products, applications, cell lines and manufacturers. The database is accessible free of charge in HTML format and as PDF download, the informations are checked and updated twice a year. Problems concerning serum free cell culture are discussed at and comments are welcome. PMID- 16256303 TI - Murine segmental duplications are hot spots for chromosome and gene evolution. AB - Mouse and rat genomic sequences permit us to obtain a global view of evolutionary rearrangements that have occurred between the two species and to define hallmarks that might underlie these events. We present a comparative study of the sequence assemblies of mouse and rat genomes and report an enrichment of rodent-specific segmental duplications in regions where synteny is not preserved. We show that segmental duplications present higher rates of molecular evolution and that genes in rearranged regions have evolved faster than those located elsewhere. Previous studies have shown that synteny breakpoints between the mouse and the human genomes are enriched in human segmental duplications, suggesting a causative connection between such structures and evolutionary rearrangements. Our work provides further evidence to support the role of segmental duplications in chromosomal rearrangements in the evolution of the architecture of mammalian chromosomes and in the speciation processes that separate the mouse and the rat. PMID- 16256304 TI - Regulation of adrenal glucocorticoid synthesis by interleukin-10: a preponderance of IL-10 receptor in the adrenal zona fasciculata. AB - Several lines of evidence indicate that cytokines can affect adrenal function. To date most of these cytokines have been shown to be pro-inflammatory, such as interleukin (IL)-1, tumor necrosis factor (TNFalpha), and IL-6. However, we have previously shown that IL-10-/- (IL-10 knockout) mice have higher serum corticosterone levels than IL-10+/+ (wild type) mice following acute immune and physiologic stress, implying that IL-10, an anti-inflammatory cytokine, regulates glucocorticoid synthesis in a negative manner. Here, we show that IL-10 knockout mice produce more corticosterone under basal conditions as well (shown by ELISA). We further support this contention by showing that in Y-1 adrenocortical cells IL 10 inhibits steroid production (StAR) (measured by the production of the corticosterone precursor, progesterone), the expression of steroidogenic acute regulatory protein (semi-quantitative RT-PCR), as well as the activity of the proximal steroidogenic enzymes P450scc and/or 3beta-hydroxysteroid dehydrogenase (3beta-HSD) (measured by progesterone production in 22(R)-hydroxycholesterol treated cells). Interestingly, all of the above-mentioned effects of IL-10 occur through its inhibition of ACTH effects, but not by IL-10 alone. Furthermore, immunocytochemistry data shows that the region of the adrenal gland responsible for the vast majority of corticosterone synthesis, the zona fasciculata, predominantly expresses the IL-10 receptor 1 (IL-10R1), with little expression in the zona glomerulosa and reticularis. These data demonstrate that IL-10 could play an important role in the regulation of glucocorticoid biosynthesis and in maintenance of homeostasis and immunity during periods of stress. PMID- 16256305 TI - Effect of sex ratio on the immune system of Oreochromis mossambicus (Peters). AB - The social environment of fish has a crucial role to play on the immune system and hence on the overall health status. Stressors of social origin such as dominance, subordination, and fight for mate have a major impact on the immune system of fish. The present study was designed with the objective of finding the effect of sex ratio of the population on the immune system of Oreochromis mossambicus. Groups of fish were maintained for 28 days in three different sex ratios i.e., (i) all-male (ii) all-female (iii) equal male and female (mixed). The specific immune response of fish was assessed by antibody response to Aeromonas hydrophila by ELISA and bacterial agglutination assay, and to SRBC by plaque forming cell assay. Nonspecific immune mechanisms were assessed in terms of serum lysozyme activity, production of intracellular reactive oxygen species (ROS) and reactive nitrogen species (RNS) by peripheral blood leukocytes. Disease resistance against live, virulent A. hydrophila was performed to assess the overall functional immunity. The results showed that antibody responses and numbers of antibody producing cells were increased in fish in the equal male and female sex ratio group compared to fish in monosex ratio groups. Similar enhancement was also observed in nonspecific serum lysozyme level and the ROS and RNS production. The host resistance test revealed that enhanced immunity in equal male and female sex ratio group was protective against A. hydrophila infection. The study clearly reveals positive and negative effect of sex ratio on the immune system of O. mossambicus. PMID- 16256306 TI - Profiling the autoantibody repertoire by serological antigen selection. AB - The identification of disease related autoantigens targeted by pathogenic T- and B-cell responses is crucial for the development of improved therapies for autoimmune diseases. To identify immunogenic targets recognized by the humoral immune response, we have recently applied a novel and powerful molecular approach, named 'serological antigen selection' (SAS). This method involves the display of a cDNA expression library on filamentous phage and subsequent selection on patient immunoglobulin G (IgG). In the present study, we have cloned a cDNA repertoire from a multiple sclerosis (MS) patient in pVI phage display vectors and performed selections on pooled MS cerebrospinal fluid (CSF) samples immobilized with anti-human IgG. To further streamline this procedure, we report an optimized SAS procedure in which we have successfully established methods for enrichment of MS-specific candidate antigens. In conclusion, the broad applicability of the SAS method makes it a highly promising method for investigating the autoimmune repertoire. PMID- 16256307 TI - Prophylactic administration of abatacept prevents disease and bone destruction in a rat model of collagen-induced arthritis. AB - Abatacept is the first in a new class of agents that selectively modulates T-cell activation by attenuating CD28-mediated co-stimulation. This study examined the effects of abatacept on disease development in a rat model of collagen-induced arthritis (CIA). The rats were treated with either abatacept (1mg/kg) or control IgG beginning at the time of induction of CIA. By day 16, significant paw swelling was observed in IgG-treated control animals that continued to increase, reaching a plateau on day 21. Prophylactic treatment with abatacept completely abrogated paw swelling throughout the study. Histopathology demonstrated a significant reduction in inflammation, cartilage destruction, bone resorption and pannus formation. Abatacept treatment resulted in 90% inhibition of circulating collagen-specific antibodies and decreased the serum expression of many cytokines and chemokines that were upregulated in diseased animals. Immunohistochemical analysis of the ankle joints demonstrated that interleukin-6 production was reduced in the tissues and the numbers of osteoclasts present in the joints were also decreased. Ankle microcomputer tomography (micro-CT) analyses dramatically demonstrated the protective effects of abatacept on bone destruction in these animals. Data presented here demonstrate that prophylactic administration of abatacept significantly inhibits the onset and progression of disease in a rat CIA model, with reductions in inflammation, inflammatory mediators, and bone and joint destruction. PMID- 16256309 TI - The Social Occupational Functioning Scale (SOFS): a brief measure of functional status in persons with schizophrenia. AB - The social functioning of persons with schizophrenia contributes to their overall functional outcome and ability to live in the community. Enhancing the level of social functioning is an important treatment goal. The present study describes the development of the Social Occupational Functioning Scale (SOFS), a brief, yet comprehensive, easy to administer measure of social functioning for use in busy clinical settings. It has adequate psychometric properties in terms of reliability and validity. Exploratory factor analysis revealed a three-factor structure comprising of adaptive living skills, social appropriateness and interpersonal skills, accounting for 59% of the variance in total SOFS score. PMID- 16256308 TI - Predicting seizure frequency after epilepsy surgery. AB - OBJECTIVE: To identify clinical features related to seizure frequency after epilepsy surgery in patients with recurrent seizures. BACKGROUND: No studies have examined the differences between patients who have rare seizures and patients who experience frequent seizures after epilepsy surgery. Since seizure frequency correlates with morbidity and quality of life, it is desirable to know which preoperative clinical features predict postoperative seizure frequency. METHODS: Patients with recurrent seizures were placed in two categories: rare postoperative seizures (< or =2 per year) and frequent postoperative seizures (> or =12 per year) using seizure frequency in the second postoperative year. Variables included preoperative seizure frequency, age of first risk, age at first seizure, epilepsy duration, age at surgery, history of febrile convulsions, tonic-clonic seizures, status epilepticus, or family history, IQ, magnetic resonance imaging (MRI), and positron emission tomography (PET). Variables were analyzed using non-parametric tests to assess relationship to postoperative seizure frequency. RESULTS: Of 475 patients who had epilepsy surgery, 111 had rare or frequent seizures in the second postoperative year. After anterior temporal lobectomy (ATL), age of first risk< or =5 years and presence of mesial temporal sclerosis on MRI were associated with rare seizures (66% of patients), whereas lack of these risk factors was associated with frequent seizures (75% of patients) (p<0.03). For non-ATL operations, preoperative seizure frequency of > or =20 seizures per month was associated with frequent postoperative seizures (p=0.03). No other variables influenced outcome. CONCLUSIONS: Some preoperative clinical features correlate with postoperative seizure frequency in patients with recurrent seizures after epilepsy surgery. This has implications for the surgical decision making process and early postoperative management. PMID- 16256310 TI - Investigation of metamemory dysfunctions in first-episode schizophrenia. AB - A number of recent studies have suggested that schizophrenia patients share metamemory deficits, particularly, a decreased ability to distinguish between errors and correct responses in terms of response confidence (i.e., decreased confidence gap): patients are over-confident in errors while at the same time being under-confident in responses that are in fact correct. This, along with increased error rates, leads to an inflation of inaccurate but confidently held memories, which has been termed knowledge corruption. Previous studies on metamemory in schizophrenia patients predominantly tested chronic patients, leaving open the possibility that metamemory deficits stem partly from increased chronicity and long-term treatment. The primary aim of the current study was to establish whether a decreased confidence gap is also detectable in first-episode schizophrenia. For this purpose, a source memory task was administered to 30 first-episode patients with a diagnosis of schizophrenia or schizophreniform disorder, and 15 healthy control subjects. During encoding, items were read aloud by the experimenter and the participant in alternating order. For the recognition phase, participants were required to state the source of the item, and their confidence in their response. In agreement with previous studies, the patients displayed a decreased confidence gap, and increased knowledge corruption relative to controls. A reduced distinction between correct and incorrect information in metacognition is proposed to be a vulnerability factor for the development of delusions in schizophrenia. PMID- 16256311 TI - Expression of split dnaE genes and trans-splicing of DnaE intein in the developmental cyanobacterium Anabaena sp. PCC 7120. AB - Protein intein is widespread in a variety of organisms. Several intein elements are also present in cyanobacteria, and some of them have been studied biochemically in vitro. However, no evidence is available for intein removal in vivo in cyanobacteria. In the filamentous cyanobacterium Anabaena sp. strain PCC 7120, the DNA replication factor DnaE is encoded by two split open reading frames (ORFs) far apart from each other on the chromosome, and each of them could contain a split intein element. This organism can undergo a developmental process leading to the formation of nitrogen-fixing cells, or heterocysts. Heterocysts are terminally differentiated cells with arrest of cell cycle. Since DnaE is an important cell cycle element involved in DNA replication, we would like to provide in vivo evidence for DnaE intein removal in cyanobacteria and determine whether mature DnaE protein is still present in heterocysts. In this study, we showed that the products of these two ORFs were joined together to form a complete DnaE protein through the process of protein trans-splicing. More interestingly, protein trans-splicing could be detected in vivo for the first time in cyanobacteria, which allowed us to compare the formation of mature DnaE protein in heterocysts and vegetative cells, and show that mature DnaE protein could be formed in both cell types. Transcriptional fusion between the promoter regions of the two split ORFs and gfp reporter also demonstrate that both ORFs are transcribed in vegetative cells and heterocysts, without strong variation during the process of heterocyst differentiation. Although heterocysts are terminally differentiated and may not replicate its chromosome, the expression and maturation of DnaE in these cells may underlie the need for DNA replication machinery in processes such as DNA recombination and repair. PMID- 16256312 TI - Purification of a new manganese peroxidase of the white-rot fungus Irpex lacteus, and degradation of polycyclic aromatic hydrocarbons by the enzyme. AB - The white-rot fungus Irpex lacteus has been reported to be an efficient degrader of polycyclic aromatic hydrocarbons, polychlorinated biphenyls and pentachlorophenol. The fungus produces ligninolytic enzymes laccase, lignin peroxidase and manganese peroxidase (MnP), the latter being the major one produced. MnP was purified using anion exchange and size exclusion chromatography. SDS-PAGE showed the purified MnP to be a monomeric protein of 37 kDa (37.5 kDa using MALDI-TOF) with an isoelectric point at 3.55. The pH optimum was relatively broad, from 4.0 to 7.0 with a peak at pH 5.5. Kinetic constants K(m) were 8 microM for H(2)O(2) and 12 or 31 microM for Mn(2+) depending on the substrate. The enzyme did not perform oxidation in the absence of H(2)O(2) or Mn(2+). MnP was active at 5-70 degrees C with an optimum between 50-60 degrees C. At temperatures above 65 degrees C the enzyme rapidly lost activity. Degradation of four representatives of PAHs (phenanthrene, anthracene, fluoranthene, and pyrene) was tested and the enzyme showed the ability to degrade them in vitro. Major degradation products of anthracene were identified. The results confirm the role of MnP in PAH degradation by I. lacteus, including cleavage of the aromatic ring. PMID- 16256313 TI - Two heterogeneously vancomycin-intermediate clinical isolates of methicillin sensitive and methicillin-resistant Staphylococcus aureus in a Turkish university hospital: brief report of a surveillance study. PMID- 16256314 TI - Magnetic resonance imaging in patients with obsessive-compulsive disorder with good versus poor insight. AB - The DSM-IV provides two subtypes of obsessive-compulsive disorder (OCD), labelled as OCD with insight and OCD with poor insight. For the latter, patients generally fail to recognize that the obsessions or compulsions are excessive or unreasonable. Several studies have shown significant brain abnormalities in OCD patients. However, at present, it remains unclear whether a specific pattern of structural brain abnormalities is related to poor insight in OCD. In the present study, magnetic resonance imaging (MRI) findings were compared in OCD patients with insight versus those with poor insight. Outpatients with diagnoses of OCD according to DSM-IV (300.30) and ICD-10 (F42) (n = 84; mean age 38+/-13; 35 females, 49 males) were dichotomized into the two subtypes. All subjects underwent an MRI examination. MRI findings were rated as "MRI abnormality" and "normal MRI." In our sample, 48% of the patients had MRI abnormalities. There was a highly significant difference between the two groups according to frequencies of MRI abnormalities, with 83% of the patients with poor insight showing MRI abnormalities compared with only 21% of the patients with insight. The specifier "poor insight" helps to identify a subgroup of OCD with a higher frequency of brain abnormalities of various types. This distinction should be taken into account in future studies concerning the course and therapeutic outcome of OCD. PMID- 16256315 TI - Dorsolateral prefrontal cortex morphology and short-term outcome in first-episode schizophrenia. AB - Several clinical predictors of outcome in schizophrenia have been described; however, very few studies have examined neurobiological factors that predict outcome. The objective of this study was to examine the value of the morphology of the dorsolateral prefrontal cortex (DLPFC) determined by magnetic resonance imaging (MRI) as a predictor of short-term functional outcome in antipsychotic naive patients with first-episode schizophrenia or schizoaffective disorder. Twenty-seven consecutive patients with first-episode schizophrenia or schizoaffective disorder underwent structural MRI at baseline. Functional outcome - a composite measure of social and employment subscales of the Strauss-Carpenter Scale - was assessed at 1 and 2 years. Volume of the left DLPFC at baseline predicted functional outcome in schizophrenia at 1 but not at 2 years. Specific cognitive functions regulated by the left DLPFC may be critical for functional outcome. PMID- 16256316 TI - Solid-state NMR structural studies of the fibril form of a mutant mouse prion peptide PrP89-143(P101L). AB - The peptide fragment 89-143 of the prion protein (carrying a P101L mutation) is biologically active in transgenic mice when in a fibrillar form. Injection of these fibrils into transgenic mice (expressing full length PrP with the P101L mutation) induces a neurodegenerative prion disease (Kaneko et al., J. Mol. Biol. 295 (2000) 997). Here we present solid-state NMR studies of PrP(89-143)(P101L) fibrils, probing the conformation of residues in the hydrophobic segment 112-124 with chemical shifts. The conformations of glycine residues were analyzed using doubly (13)C=O labeled peptides by two-dimensional (2D) double-quantum correlation, and double-quantum filtered dephasing distance measurements. MQ-NMR experiments were carried out to probe the relative alignment of the individual peptides fibrils. These NMR studies indicate that the 112-124 segment adopts an extended beta-sheet conformation, though not in a parallel, in register alignment. There is evidence for conformational variability at Gly 113. DQ correlation experiments provide useful information in regions with conformational heterogeneity. PMID- 16256317 TI - Indirect detection of selective nuclear spin-spin interactions in a hostile environment. AB - We present a number of techniques which may be used to obtain precise values of selective spin-spin interactions between two nuclear spins in a hostile environment. Such an environment may be characterized by very fast relaxation and decoherence, e.g. due to the strong coupling of the two spins of interest to electron spins in their vicinity as well as other nuclei. Here, we used dilute paramagnetic Ce(3+) centers hosted in a single crystal of CaF(2). Selected (19)F internuclear interactions were measured indirectly by applying different electron nuclear double resonance (ENDOR) pulse sequences. PMID- 16256318 TI - Paying attention to saccadic intrusions. AB - Fixation to a target in primary gaze is invariably interrupted by physiological conjugate saccadic intrusions (SI). These small idiosyncratic eye movements (usually <1 degrees in amplitude) take the form of an initial horizontal fast eye movement away from the desired eye position, followed after a variable duration by a return saccade or drift. As the aetiology of SI is still unclear, it was the aim of this study to investigate whether SI are related to exogenous or endogenous attentional processes. This was achieved by varying (a) the "bottom up" target viewing conditions (target presence, servo control of the target, target background, target size) and (b) the 'top-down' attentional state (instruction change--'look' or 'hold eyes steady' and passive fixation versus active--'respond to change' fixation) in 13 subjects (the number of participants in each task varied between 7 and 11). We also manipulated the orientation of pure exogenous attention through a cue-target task, during which subjects were required to respond to a target, preceded by a non-informative cue by either pressing a button or making a saccade towards the target. SI amplitude, duration, frequency and direction were measured. SI amplitude was found to be significantly higher when the target was absent and SI frequency significantly lower during open loop conditions. Target size and background influenced SI behaviour in an idiosyncratic manner, although there was a trend for subjects to exhibit lower SI frequencies and amplitudes when a patterned background was present and larger SI amplitudes with larger target sizes. SI frequency decreased during the "hold eyes steady" passive command as well as during active fixation but SI direction was not influenced by the exogenous cue-target task. These results suggest that SI are related to endogenous rather than exogenous attention mechanisms. Our experiments lead us to propose that SI represent shifts in endogenous attention that reflect a baseline attention state present during laboratory fixation tasks and may prove to be a useful tool to explore higher cortical control of fixation. PMID- 16256319 TI - Alcohol consumption impairs stimulus- and error-related processing during a Go/No Go Task. AB - Alcohol consumption has been shown to increase the number of errors in tasks that require a high degree of cognitive control, such as a go/no-go task. The alcohol related decline in performance may be related to difficulties in maintaining attention on the task at hand and/or deficits in inhibiting a prepotent response. To test these two accounts, we investigated the effects of alcohol on stimulus- and response-locked evoked potentials recorded during a go/no-go task that involved the withholding of key presses to rare targets. All participants performed the task prior to drinking and were then assigned randomly to either a control, low-dose, or moderate-dose treatment. Both doses of alcohol increased the number of errors relative to alcohol-free performance. Success in withholding a prepotent response was associated with an early-enhanced stimulus-locked negativity at inferior parietal sites, which was delayed when participants failed to inhibit the motor command. Moreover, low and moderate doses of alcohol reduced N170 and P3 amplitudes during go, no-go, and error trials. In comparison with the correct responses, errors generated large response-locked negative (Ne) and positive (Pe) waves at central sites. Both doses of alcohol reduced the Ne amplitude whereas the Pe amplitude decreased only after moderate doses of alcohol. These results are consistent with the interpretation that behavioral disinhibition following alcohol consumption involved alcohol-induced deficits in maintaining and allocating attention thereby affecting the processing of incoming stimuli and the recognition that an errant response has been made. PMID- 16256320 TI - Visual magnocellular and structure from motion perceptual deficits in a neurodevelopmental model of dorsal stream function. AB - Williams syndrome (WS) is a neurodevelopmental disorder of genetic origin that has been used as a model to understand visual cognition. We have investigated early deficits in the afferent magnocellular pathway and their relation to abnormal visual dorsal processing in WS. A spatiotemporal contrast sensitivity task that is known to selectively activate that pathway was used in six WS subjects. Additionally, we have compared visual performance in 2D and 3D motion integration tasks. A novel 3D motion coherence task (using spheres with unpredictable axis of rotation) was used in order to investigate possible impairment of occipitoparietal areas that are known to be involved in 3D structure from motion (SFM) perception. We have found a significant involvement of low-level magnocellular maps in WS as assessed by the contrast sensitivity task. On the contrary, no significant differences were observed between WS and the control groups in the 2D motion integration tasks. However, all WS subjects were significantly impaired in the 3D SFM task. Our findings suggest that magnocellular damage may occur in addition to dorsal stream deficits in these patients. They are also consistent with recently described genetic and neuroanatomic abnormalities in retinotopic visual areas. Finally, selective SFM coherence deficits support the proposal that there is a specific pathway in the dorsal stream that is involved in motion processing of 3D surfaces, which seems to be impaired in this disorder. PMID- 16256321 TI - Direction and distance deficits in path integration after unilateral vestibular loss depend on task complexity. AB - The effects of peripheral vestibular disorders on the direction and distance components of the internal spatial representation were investigated. The ability of Meniere's patients to perform path integration was assessed in different situations aimed at differentiating the level of spatial processing (simple versus complex tasks), the available sensory cues (proprioceptive, vestibular, or visual conditions), and the side of the path (towards the healthy versus the lesioned side). After exploring two legs of a triangle, participants were required either to reproduce the exploration path, to follow the reverse path, or to take a shortcut to the starting point of the path (triangle completion). Patients' performances were recorded before unilateral vestibular neurotomy (UVN) and during the time-course of recovery (1 week and 1 month) and were compared to those of matched control subjects tested at similar time intervals. Both the angular and linear path components of the trajectory were impaired for patients compared to controls. However, deficits were restricted to the complex tasks, which required a higher level of spatial processing. Most deficits were maximal 1 week after UVN, and some remained up to the first post-operative month. Spatial representation was differentially impaired according to the available sensory cues: deficits were absent in active locomotor blindfolded condition, appeared in conditions involving visual and vestibular information, and were maximal when visual cues alone were available. Finally, concerning the side of the path, unilateral vestibular loss led to global impairment of the internal spatial representation, yet some asymmetrical spatial performances were observed 1 week after UVN. On the whole, results suggest that the environment experienced by the patients is different after UVN and that a different internal spatial representation is constructed, especially for tasks requiring high levels of spatial processing. PMID- 16256322 TI - Effect of surface hydrophobicity on the adhesion of S. cerevisiae onto modified surfaces by poly(styrene-ran-sulfonic acid) random copolymers. AB - The hydrophobicity of solid surfaces has been regarded as a controlling factor in microbial adhesion phenomena. In this study, the surface hydrophobicity was modified by coating with a poly(styrene-ran-sulfonic acid) random copolymer (PS-x SA, charge density (x): 0-15.3%), and the adhesion rate, J0, of S. cerevisiae performed with a direct observation technique. The results indicated that the degree of sulfonation of PS-x-SA greatly influenced the hydrophobicity of substrates and the adhesion of yeast cells. The J0 of PS-x-SA substrates were gradually decreased as increasing charge density. The interactions between cells and substrates explained by the XDLVO theory, predicted that the decrease of J0 as increasing charge density was not due to the increase of electric double layer repulsion, but mainly due to the hydrophobic acid-base interactions. Also, it predicted that microbial adhesions of PS-x-SA were mostly reversible, while some of PS and PS-5.1-SA adhered cells were hardly removed. Based on these results, XDLVO theory was effective for predicting adhesion phenomena of S. cerevisiae onto the PS-x-SA-coated substrates. PMID- 16256323 TI - Wireless on-line electromyography in recording neck muscle function: a pilot study. AB - AIM: The surface electromyography (SEMG) records muscle activity without causing pain and therefore it can be used in the on-line kinetic evaluation of rehabilitation methods. The cables are a problem in the clinical work. The aim of this pilot study was to introduce a wireless on-line SEMG method in the measurements of neck muscle activity and preliminarily evaluate its usability in dynamic exercises in a healthy and a neck pain subject. METHOD: The bilateral wireless on-line SEMG measurements were performed at the level C4-C5 of the cervical spine. The subjects (one healthy 32-year-old female and one neck pain patient, 35-year-old female) were performing slow cervical flexions with eyes open. The angle of the flexion was measured with inclinometer. RESULTS: In healthy subjects, the SEMG curve demonstrated clear flexion-relaxation rhythm whereas in the patient the rhythm was blurred. The SEMG signal quality was, however, good in both cases. CONCLUSION: Wireless on-line SEMG method seems promising in daily clinical work for the measurements of neck muscle activity. PMID- 16256324 TI - [Management of an intensive care room in the department of pediatric emergency: interest of a written protocol and a follow up chart for materiovigilance declaration]. PMID- 16256325 TI - Future perspectives in COPD. AB - The pathophysiology of chronic obstructive pulmonary disease (COPD) is complex. The development of a multidimensional index--such as the BODE index--provides a means of classifying patients with COPD that also correlates with their prognosis. The individual components of the BODE index--body mass index (B), airflow obstruction (O) dyspnoea (D) and exercise capacity (E)--incorporate the pulmonary as well as the systemic effects seen in patients with COPD. Recent research has focussed on examining these impairments (including those of metabolism and inflammation) more carefully, and determining the effects of treatment on both the systemic and physiological aspects of COPD. Ongoing research initiatives by the public and private sector will contribute to our understanding of the disease processes underlying COPD, our understanding of the benefits associated with commonly used pharmacotherapies, as well as laying the foundations for the development of new agents and therapeutic tools. Advances in the use of pharmacotherapy have been mirrored by research to better define the benefits associated with pulmonary rehabilitation. Many questions remain to be answered, but a comprehensive approach is now considered essential to the life long management of COPD, and will undoubtedly reduce the considerable socio economic burden of COPD. PMID- 16256326 TI - Utilization of date stones for production of activated carbon using phosphoric acid. AB - Date stone wastes have been utilized for production of activated carbon by chemical activation with phosphoric acid using a fluidized-bed reactor. The effects of the activation time, activation temperature, impregnation ratio, and particle size on the yield and the adsorptive capacity towards iodine were studied. The yield and the quality of the activated carbon prepared by using H3PO4 were compared with that prepared from date stones using the same equipment, and under similar conditions by using ZnCl2 as an oxidizing agent. The maximum value of the iodine number of the activated carbon produced using H3PO4 in this work was about 495 under the following conditions: impregnation ratio 0.4, activation time 60 min, activation temperature 800 degrees C, particle size 0.60 mm. The iodine number for the produced activated carbon was higher when phosphoric acid was used, compared to that when zinc chloride was used as impregnation reagent; however, the yield obtained when H3PO4 was used was lower than the yield when ZnCl2 was used. The iodine number increases significantly with increasing the activation temperature. By increasing the impregnation ratio at the same temperature, the iodine number decreased sharply and an oscillation is noticed for all the cases but it was clearer at 800 degrees C. The average variation of the iodine number for the whole range of particle size used in this work is +/-10%. PMID- 16256327 TI - Development of rice husks-plastics composites for building materials. AB - In this paper, a new effective recycling method for rice husks and waste expanded polystyrene is developed by using a combination of both wastes. A styrene solution of waste expanded polystyrene is used as a binder for rice husks plastics composites. The composites are prepared with various mix proportions by a hot press molding method, and tested for apparent density, water absorption, expansion in thickness, and dry and wet flexural strengths. From the test results, the apparent density of the composites is increased with increasing binder content and filler-binder ratio. Their flexural strength and wet flexural strengths reach maximums at a binder content of 30.0% and a filler-binder ratio of 1.0. Their water absorption and expansion in thickness are decreased with increasing binder content and filler-binder ratio. Since the composites have a high flexural strength and water resistance, their uses as building materials are expected. PMID- 16256328 TI - dc-Sheet resistance as sensitive monitoring tool of protein immobilization on thin metal films. AB - The suitability of high resolution, in situ dc-sheet resistance monitoring (SRM) as a simplified and reliable sensing technique towards detection and tracking of protein immobilization has been explored. Non-specific adsorption of bovine serum albumin (BSA) onto a very thin gold film, acting as the sensing resistor, has been employed as a model system. For comparison, the novel sensing method was combined with surface plasmon resonance (SPR) spectroscopy, using the same flow cell and sensing surface. Two different, well known adsorption states, involving a composite layer of irreversibly and reversibly bound BSA, were clearly resolved by both methods. Clearly structured, pronounced and fully reproducible film resistance modulations have been resolved in the associated SRM data. The transition from reversibly bound BSA to the diluted protein phase is associated with an unusually large decrease in the dc-sheet resistance. The observed resistance modulation magnitude for an adsorbed BSA monolayer corresponds to approximately 1%, and up to 100 mOmega at a 10 Omega sensing resistor. The sheet resistance of irreversibly bound BSA was determined to 0.24 kOmega/cm2, and the associated specific resistivity estimated to 1-2x10(4) Omega cm. PMID- 16256330 TI - Combined low-dose spinal-epidural anesthesia versus single-shot spinal anesthesia for elective cesarean delivery. AB - Combined spinal-epidural anesthesia balancing low-dose intrathecal bupivacaine/fentanyl and low-dose epidural bupivacaine may be more useful than single-shot spinal anesthesia for cesarean delivery in reducing incidences of adverse effects such as hypotension and nausea and in shortening motor recovery. Combined spinal-epidural anesthesia (n=50) or spinal anesthesia (n=50) was randomly performed in 100 parturients. Intrathecal bupivacaine 6 mg added by fentanyl 20 mug followed after 5 min by 10 mL of 0.25% epidural bupivacaine were used for combined spinal-epidural and intrathecal bupivacaine 9 mg with fentanyl 20 mug for spinal anesthesia. The initial sensory block level was higher in the spinal group (P<0.001), although the maximum levels were the same (T3). Complete surgical anesthesia was achieved and no patient complained of intraoperative pain in either group. Patients in the spinal group had denser motor block in the extremities and a higher incidence of hypotension (P<0.05) and nausea and vomiting (P<0.05). Motor recovery was faster in the combined spinal-epidural group (P<0.001). We concluded that combined spinal-epidural anesthesia using low dose local anesthetic-opioid spinal anesthesia and routine epidural supplementation before surgery had some potential advantages over single-shot spinal anesthesia in the lower incidences of adverse effects and quicker recovery. PMID- 16256329 TI - Biocatalysis in semi-aqueous and nearly anhydrous conditions. AB - In the past few years there have been prolific advances in activating enzymes for nonaqueous biocatalysis. Molecular dynamics simulations complement recent experimental results and offer new insights into the deleterious effects of organic solvents, such as water stripping and active-site penetration. Methods for activating enzymes in semi-aqueous or nonaqueous media include protein engineering, chemical modification, and co-lyophilization with non-buffer salts. Enzyme immobilization on novel polymeric supports and the use of zeolite molecular sieves can also increase solvent tolerance, enhance activity, and improve enantioselectivity. The recent implementation of enzymes in ionic liquids has also led to better long-term stability relative to traditional organic solvents and the simultaneous solubilization of enzymes, cofactors and substrates. PMID- 16256331 TI - Implications for anaesthesia in a patient established on clozapine treatment. AB - Clozapine is an atypical antipsychotic agent with a novel pharmacological profile and multiple clinical properties. Because of its side effects, it is recommended in treatment of severe resistant schizophrenia for which purpose it is remarkably effective. Little is known about the safety profile of clozapine during pregnancy and labour and because it is now used more commonly to manage schizophrenia, it is important that we as anaesthetists are aware of its many interactions and potential side effects. We present a case of a successful emergency caesarean section in a schizophrenic patient on clozapine treatment. PMID- 16256332 TI - Use of an information leaflet to improve general practitioners' knowledge of post dural puncture headache. AB - BACKGROUND: The number of women receiving neuraxial anaesthesia for labour and delivery is increasing. Women are also being discharged into the community sooner after delivery. Thus, complications arising from neuraxial anaesthesia may present to general practitioners, so it is of vital importance that they are familiar with and can manage potential problems associated with these anaesthetic techniques. METHODS: A questionnaire was sent to 126 local general practitioners to discover their knowledge of the symptoms, diagnosis and treatment of post dural puncture headache in the parturient. An information leaflet was then circulated to all general practitioners in the region, detailing headaches and other potential problems following epidural analgesia for childbirth, and the questionnaire was reissued. RESULTS: The first questionnaire demonstrated that they had poor knowledge of the symptoms, diagnosis and treatment of post dural puncture headache in the parturient. Following the dissemination of the information leaflet, responses to the second questionnaire showed a significant improvement. CONCLUSION: By comparing the two sets of answers, we demonstrated that the leaflet has successfully improved knowledge of post dural puncture headache and other potential sequelae of obstetric epidural analgesia among general practitioners. PMID- 16256333 TI - Does postdural puncture headache left untreated lead to subdural hematoma? Case report and review of the literature. AB - The patient was a 39-year-old pregnant woman who was scheduled for cesarean section. Spinal anesthesia was induced using a 26-gauge needle with an atraumatic bevel. Postoperatively, the patient developed cranial subdural hematoma manifesting as severe non-postural headache, associated with right eye tearing, fifth cranial nerve palsy and left hemiparesis. The diagnosis was confirmed by computed tomography scan. The patient was managed by careful neurological follow up associated with conservative treatment and recovered fully after 12 weeks. Our report reviews the literature on 46 patients who developed a postdural puncture headache complicated by subdural hematoma following spinal or epidural anesthesia. It is possible that postdural puncture headache left untreated may be complicated by the development of subdural hematoma. Patients developing a postdural puncture headache unrelieved by conservative measures, as well as the change from postural to non-postural, require careful follow-up for early diagnosis and management of possible subdural hematoma. PMID- 16256334 TI - Height of confusion: assessing regional blocks before caesarean section. PMID- 16256335 TI - There is no place in modern obstetrics for racemic bupivacaine. PMID- 16256336 TI - Inadvertent administration of magnesium sulfate through the epidural catheter: report and analysis of a drug error. AB - We present two reports of pregnant women in labor who inadvertently received a magnesium sulfate solution in their epidural space. Both women received approximately 9 mg of magnesium sulfate, and neither of them demonstrated any signs or symptoms of focal neurological toxicity. Once the mistakes were discovered and appropriate medication was delivered, the patients attained an acceptable level of analgesia. PMID- 16256337 TI - Comparison of continuous background infusion plus demand dose and demand-only parturient-controlled epidural analgesia (PCEA) using ropivacaine combined with sufentanil for labor and delivery. PMID- 16256338 TI - Preoperative anxiety and postoperative satisfaction in women undergoing elective caesarean section. AB - BACKGROUND: The primary aim was to investigate whether preoperative anxiety in women undergoing elective caesarean section predicts postoperative maternal satisfaction with the process, perceptions of recovery, analgesic use or length of hospital stay. Other factors that might influence postoperative satisfaction were also explored. METHOD: In 85 women awaiting elective caesarean section, anxiety, social support and aspects of preparation were measured in the 24 hours preceding surgery. Maternal satisfaction and perceptions of recovery were assessed around the third postoperative day. Satisfaction with the preoperative information from the anaesthetist and postoperative pain relief were also measured at this time. Medical notes were used to gather information on analgesia use and length of hospital stay. RESULTS: Preoperative anxiety scores were comparable with those of general surgical/medical patients. Preoperative trait anxiety and state anxiety were inversely associated with postoperative maternal satisfaction. State anxiety was also inversely associated with better recovery. Preoperative anxiety was not associated with analgesic use or length of hospital stay. Linear regression analysis indicated the degree of satisfaction with information from the anaesthetist and perceived emotional support from the partner explained 52% of the variance in postoperative maternal satisfaction. CONCLUSION: Lower preoperative anxiety is associated with greater maternal satisfaction with elective caesarean section and better recovery. Information provided by anaesthetists and perceived emotional support are also of importance. It may be possible to identify women with high anxiety and facilitate satisfaction and recovery through providing additional supportive input. PMID- 16256339 TI - There is no place in modern obstetrics for racemic bupivacaine. PMID- 16256340 TI - Arm and ankle blood pressure during caesarean section. AB - BACKGROUND: We have previously reported that measurement of non-invasive blood pressure during caesarean section under spinal anaesthesia fails in over 50% of cases. We felt that errors would be less likely if blood pressure could be measured at the ankle as it is immobile during caesarean section. The purpose of our study was to determine whether blood pressure measurement at the ankle was equivalent to the arm. METHOD: Following ethical approval, informed consent was obtained from 30 women scheduled for elective caesarean section. Two non-invasive blood pressure cuffs, one on the upper arm and one on the ankle, were used to measure blood pressures at three timed intervals: before spinal insertion, before surgery and after delivery of the neonate. RESULTS: Using the method of Bland and Altman we found that there was only marginal agreement between the two methods. On eight out of 15 occasions where there was a greater than 20% fall in arm systolic blood pressure, this was not detected at the ankle. CONCLUSION: We cannot recommend the use of the ankle to measure blood pressure during caesarean section. PMID- 16256341 TI - Evaluation of the platelet function analyzer (PFA-100) vs. the thromboelastogram (TEG) in the parturient. AB - BACKGROUND: The platelet function analyzer (PFA-100) is a bedside test of coagulation designed to evaluate platelet function. It measures the time required for whole blood to occlude a membrane impregnated with either epinephrine (EPI) or adenosine 5'diphosphate (ADP). The results are reported as closure time (CT EPI or CT-ADP) in seconds. The thromboelastogram (TEG) measures whole blood clotting and the maximum amplitude (MA) correlates with platelet count and function. We wished to establish whether there is a correlation between the CT and platelet count, between the CT and MA, and between the MA and platelet count. METHODS: Platelet count, CT, and MA were measured in blood drawn from 172 healthy term parturients using the PFA-100. RESULTS: We were unable to detect a significant correlation between the CT-EPI and platelet count (r=-0.1, P=0.21), or the CT-ADP and platelet count (r=-0.02, P=0.83). We also did not find a correlation between the CT-EPI and MA (r=-0.13, P=0.12) or between the CT-ADP and MA (r=-0.11, P=0.19). However, we found a significant correlation between platelet count and MA (r=0.33, P<0.001). CONCLUSIONS: We conclude that the CT does not correlate with the platelet count or MA in the parturient, but the TEG does. Therefore the TEG may be a better tool to evaluate coagulation in the parturient with thrombocytopenia. PMID- 16256342 TI - Prognostic relevance of uPAR-del4/5 and TIMP-3 mRNA expression levels in breast cancer. AB - Recently, two components of important protease systems in cancer, i.e., the urokinase plasminogen activator receptor (uPAR) mRNA splice variant uPAR-del4/5 and the tissue inhibitor of matrix metalloproteinase-3 (TIMP-3), were independently reported to be of prognostic value in breast cancer. In the present study, we have evaluated the impact of both these factors on disease-free survival (DFS) in 205 breast cancer patients by assessing mRNA expression in tumour tissue by quantitative PCR. High uPAR-del4/5 mRNA expression was associated with shorter DFS in breast cancer patients (P=0.0363), whereas high TIMP-3 mRNA levels were associated with a good prognosis (P=0.0049). Furthermore, by combining uPAR-del4/5 with TIMP-3 values, we demonstrate that breast cancer patients with high uPAR-del4/5 and low TIMP-3 mRNA levels had a highly significantly shorter DFS in comparison to those patients with low uPAR-del4/5 and high TIMP-3 mRNA expression (P<0.0001). These patients had a more than 6-fold higher risk for disease recurrence or death in multivariate analysis. Therefore, considering the prognostic impact of two proteolytic factors stemming from complementary protease systems may improve the prediction of disease recurrence in breast cancer. PMID- 16256343 TI - Partial characterization of extracellular polysaccharides from cyanobacteria. AB - Four cyanobacterial strains, Cyanothece sp., Oscillatoria sp., Nostoc sp. and Nostoc carneum were studied for physico-chemical characterization of extracellular polysaccharide (EPS) secreted during the controlled growth condition. Hydrolyzed EPSs showed the compositional involvement of four sugar moieties viz. mannose, glucose, xylose and ribose in varying combinations. Infrared spectra of EPSs showed a specific absorbance of O-H stretching at 3448 3400 cm(-1), asymmetrical-symmetrical C-H stretching at 2924 and 2854 cm(-1) and a bending vibration of C-H at 1400-1380 cm(-1). Absorbance at 1259 and 1140 cm( 1) with Cyanothece sp. EPS, indicated the presence of sulfur containing functional group. Thermal gravimetric analysis and differential scanning calorimetric analysis confirmed the polysaccharides thermal stability as high as around 250 degrees C. In the presence of 0.1 M NaCl aqueous solution, the intrinsic viscosity of polysaccharides from Oscillatoria sp. and Nostoc sp. decreased 1.6 fold, whereas, 3-5 fold reduction in intrinsic viscosity was observed with commercially available guar and xanthan gum. PMID- 16256345 TI - Phytoremediation of chromium by model constructed wetland. AB - Chromium is a pollutant present in tannery wastewater, its removal is necessary for protection of the environment. Penisetum purpureum, Brachiaria decumbens and Phragmites australis were grown hydroponically in experimental gravel beds to determine their potential for the phytoremediation of solutions containing 10 and 20 mg Cr dm(-3). These concentrations, similar to tannery wastewater after initial physico-chemical treatment were used with the aim of developing an economic secondary treatment to protect the environment. All the systems achieved removal efficiencies of 97-99.6% within 24 h. P. purpureum and B. decumbens removed 78.1% and 68.5% respectively within the first hour. Both P. purpureum and B. decumbens were tolerant of the concentrations of chromium applied, but P. purpureum showed the greatest potential because its faster growth and larger biomass achieved a much greater chromium removal over the whole length of time of the experiment. PMID- 16256344 TI - Evaluation of the mechanical, physical properties and decay resistance of particleboard made from particles impregnated with Pinus brutia bark extractives. AB - The mechanical, physical properties and decay resistances of particleboard made from particles impregnated with Pinus brutia bark extractives were examined. Properties included were modulus of rupture, modulus of elasticity, internal bond, thickness swelling, and weight loss according to European standards. The results showed that particleboards made from particles impregnated with bark extractives had significantly lower mechanical values than those made from unimpregnated particles. Impregnating wood particles with bark extractives improved the decay resistance and thickness swelling of particleboard. Increasing concentration of the extractives decreased the mechanical properties and improved the thickness swelling and decay resistance of the panels. Particleboards made from 1% P. brutia bark extractives met the specifications for modulus of rupture and internal bond strength for general purposes. PMID- 16256346 TI - Investigating the suitability of the Calgary Biofilm Device for assessing the antimicrobial efficacy of new agents. AB - This study investigated the suitability of the Calgary Biofilm Device (CBD), originally designed as a test surrogate for indwelling medical devices, for assessing the efficacy of antimicrobials developed for food and food contact surface disinfection applications. The conditions for the development of uniform biofilms from pure and mixed bacterial cultures of wild type Escherichia coli and Listeria innocua were optimized. We were able to recover approximately 2 x 10(6) colony forming units (CFU) from the biofilms formed on the individual pegs of the device in 24 h. Further, the parameters for the consistent release of the cells from the biofilms were optimized; test showed that the number of cells released was uniform and reproducible. The consistency and reproducibility of the biofilms formed on the pegs was evaluated using scanning electron microscopy and by plate count method. The efficacies of disinfectants on cells residing in biofilms versus planktonic cells were compared. For both species, higher concentrations of disinfectants were needed to eliminate attached cells as compared with planktonic cells. This study establishes the value of the CBD for generating consistent biofilms from either pure or mixed cultures. These biofilms can be used to assess efficacies of disinfectants against cells that have colonized the surfaces of foods and food-processing equipment. Such a system could serve as a standard surrogate for evaluating new disinfectants designed to reduce or eliminate biofilms from food-contact surfaces. PMID- 16256347 TI - Biosorption characteristics of Aspergillus flavus biomass for removal of Pb(II) and Cu(II) ions from an aqueous solution. AB - The Pb(II) and Cu(II) biosorption characteristics of Aspergillus flavus fungal biomass were examined as a function of initial pH, contact time and initial metal ion concentration. Heat inactivated (killed) biomass was used in the determination of optimum conditions before investigating the performance of pretreated biosorbent. The maximum biosorption values were found to be 13.46 +/- 0.99 mg/g for Pb(II) and 10.82 +/- 1.46 mg/g for Cu(II) at pH 5.0 +/- 0.1 with an equilibrium time of 2 h. Detergent, sodium hydroxide and dimethyl sulfoxide pretreatments enhanced the biosorption capacity of biomass in comparison with the heat inactivated biomass. The biosorption data obtained under the optimum conditions were well described by the Freundlich isotherm model. Competitive biosorption of Pb(II) and Cu(II) ions was also investigated to determine the selectivity of the biomass. The results indicated that A. flavus is a suitable biosorbent for the removal of Pb(II) and Cu(II) ions from aqueous solution. PMID- 16256348 TI - A process to prepare a synthetic filter material containing nutrients for biofiltration. AB - In this study, an optimal process to prepare a synthetic filter material (poly(vinyl alcohol) (PVA)/peat/KNO(3) composite bead) containing nutrients was developed for biofiltration. The optimal preparing condition was that each of the peat and PVA aqueous solutions contains 6.4 g KNO(3) and the nitrogen content in the boric and phosphate aqueous solutions must retain higher than 3.94 and 1.52 g N/l, respectively. The equilibrium amount of water-soluble nitrogen dissolved out of the prepared composite bead was between 7.95 and 8.21mg N/g dry solid. The path of water-soluble nitrogen dissolving out of the A-type bead was the water soluble nitrogen dispersed in the peat phase initially diffused into the outer PVA phase and then it diffused out of the bead surface. And the path of water soluble nitrogen dissolving out of the H-type bead was the water-soluble nitrogen dispersed in both the peat and PVA phases simultaneously diffused into the outer PVA phase and out of the bead surface, respectively. The microbial growth rate k(g) of the H-type composite bead was higher than that of the A-type composite bead approximately 1.09-1.58 times, and its value was between 0.100 and 0.417 day(-1) as the composite bead was immersed in 0-0.896 M KNO(3) solution. The maximum value of k(g) appeared at the composite bead immersed in 0.384 M KNO(3) solution and was higher than that of the compost by a factor approximately 1.49. The percentage of removed volatile organic compounds (VOCs) remained at more than 98% during the biofilter operating 230 days as the composite bead was immersed in KNO(3) aqueous solution before packing. This composite bed was without the further addition of nutrients during this operating period. It was proved that this composite bead was superior to the compost as a filter material. PMID- 16256349 TI - Benzylprotected aromatic phosphonic acids for anchoring peptides on titanium. AB - The development of biocompatible coatings is an ongoing issue. Mimicking the physiological adhesion process of osteoblasts to the extracellular matrix improves cell adhesion of osteoblasts in vitro and results in improved and earlier osseous integration of implants in vivo. Titanium, an often used material in implant surgery, can be easily coated by peptides bearing phosphonic acid groups. We report here, the synthesis of benzyl protected phosphonic acids suitable for solid-phase peptide synthesis (SPPS), which can be easily deprotected with TFA. PMID- 16256350 TI - The use of transcranial magnetic stimulation in the clinical evaluation of suspected myelopathy. AB - Central motor conduction time (CMCT) and motor evoked potential (MEP) latencies measured by using transcranial magnetic stimulation (TMS) are parameters used to evaluate electrophysiologic function of the corticospinal motor tract. We present 5 cases to illustrate how the use of TMS had contributed to clinical management. CMCT and MEP latency measurements were found to be useful in determining the significance of lesions seen on neuroimaging and helped clinical decisions in the presence of multiple lesions or multiple clinical conditions that cause similar clinical manifestations. TMS study is particularly useful in localizing levels of conduction defect. PMID- 16256351 TI - 3D-QSAR studies on tripeptide aldehyde inhibitors of proteasome using CoMFA and CoMSIA methods. AB - The ubiquitin-proteasome pathway plays a crucial role in the regulation of many physiological processes and in the development of a number of major human diseases, such as cancer, Alzheimer's, Parkinson's, diabetes, etc. As a new target, the study on the proteasome inhibitors has received much attention recently. Three-dimensional quantitative structure-activity relationship (3D QSAR) studies using comparative molecule field analysis (CoMFA) and comparative molecule similarity indices analysis (CoMSIA) techniques were applied to analyze the binding affinity of a set of tripeptide aldehyde inhibitors of 20S proteasome. The optimal CoMFA and CoMSIA models obtained for the training set were all statistically significant with cross-validated coefficients (q(2)) of 0.615, 0.591 and conventional coefficients (r(2)) of 0.901, 0.894, respectively. These models were validated by a test set of eight molecules that were not included in the training set. The predicted correlation coefficients (r(2)) of CoMFA and CoMSIA are 0.944 and 0.861, respectively. The CoMFA and CoMSIA field contour maps agree well with the structural characteristics of the binding pocket of beta5 subunit of 20S proteasome, which suggests that the 3D-QSAR models built in this paper can be used to guide the development of novel inhibitors of 20S proteasome. PMID- 16256352 TI - Studies of interaction of trichloro{eta2-cis-N,N-dimethyl-1-[6-(N',N'-dimethyl ammoniummethyl)-cyclohex-3-ene-1-yl]-methylammonium}platinum(II) chloride with DNA: Effects on secondary and tertiary structures of DNA. Cytotoxic assays on human ovarian cancer cell lines, resistant and non-resistant to cisplatin. AB - The studies of interaction with DNA and the cytotoxic activity of a new organometallic platinum(II) compound are presented. The ability of this new platinum complex to modify secondary DNA structure was explored by circular dichroism (CD). Electrophoretic mobility showed changes in tertiary DNA structure, and atomic force microscopy (AFM) revealed morphological changes of plasmid DNA (pBR322). This compound breaks the traditional structure-activity rules for cis-platinum compounds, but it could be of interest because of its different kinetics. An organometallic bond normally shows a trans-effect higher than that of an amine ligand, and that fact, a priori, could contribute to a higher DNA binding rate. Several ovarian cancer cell lines, resistant and non resistant to cisplatin, were exposed to increasing concentrations of cisplatin and complex 5 for 24 h, after which time the cell number/viability was determined by the colorimetric MTT assay. A lower cytotoxicity but also a lower resistant factor was observed for organometallic compound 5 than for cisplatin, against A2780 and A2780cisR cell lines. This result is consistent with the DNA interaction degree observed by the aforementioned techniques. PMID- 16256353 TI - Theoretical proton affinities of alpha1 adrenoceptor ligands. AB - A systematic study has been performed of the proton affinity of a large family of agonists and antagonists of the alpha1-adrenoceptor at the B3LYP/6-31G* level of theory. After a conformational search, all the N atoms were considered as protonation sites and protonation energy values were determined. The inclusion of solvation by means of the Onsager model yielded stabilization in the proton affinity values obtained. In addition, a good correlation was found between the proton affinity values corresponding to the first protonation in gas phase of some of the compounds and their corresponding experimental affinity constants K(i) for the alpha1A adrenergic receptor. PMID- 16256354 TI - Classification of dopamine antagonists using functional feature hypothesis and topological descriptors. AB - The designing of selective dopamine antagonists for their own subreceptors can be useful in individual therapy of various neuropsychiatric disorders. Three dimensional pharmacophore hypothesis and two-dimensional topological descriptors were used to investigate and compare different classes of dopamine antagonists. The structurally diverse D(3) and D(4) antagonists above preclinical trials were selected to map common structural features of highly selective and efficacious antagonists. The generated pharmacophore hypotheses were successfully employed as discriminative probe for database screening. To filter out the false positive from screening hits, the classification models by two-dimensional topological descriptors were built. Molconn-Z and BCUT topological descriptors were employed to develop a classification model for 1328 dopamine antagonists from MDDR database. The soft independent modeling of class analogy and artificial neural network, two supervised classification techniques, successfully classified D(1), D(3), and D(4) antagonists at the average of 80% rates into their own active classes. The mean classification rates for D(2) antagonists were obtained to 60% due to insufficient selective D(2) antagonists. In this paper, we report the validity of our models generated using functional feature hypotheses and topological descriptors. The combining both of classification using functional feature hypotheses and topological descriptors would be a useful tool to predict selective antagonists. PMID- 16256355 TI - Direct observation (NMR) of the efficacy of glucagon receptor antagonists in murine liver expressing the human glucagon receptor. AB - The demonstration of pharmacodynamic efficacy of novel chemical entities represents a formidable challenge in the early exploration of synthetic lead classes. Here, we demonstrate a technique to validate the biological efficacy of novel antagonists of the human glucagon receptor (hGCGR) in the surgically removed perfused liver prior to the optimization of the pharmacokinetic properties of the compounds. The technique involves the direct observation by (13)C NMR of the biosynthesis of [(13)C]glycogen from [(13)C]pyruvate via the gluconeogenic pathway. The rapid breakdown of [(13)C]glycogen (glycogenolysis) following the addition of 50 pM exogenous glucagon is then monitored in real time in the perfused liver by (13)C NMR. The concentration-dependent inhibition of glucagon-mediated glycogenolysis is demonstrated for both the peptidyl glucagon receptor antagonist 1 and structurally diverse synthetic antagonists 2-7. Perfused livers were obtained from a transgenic mouse strain that exclusively expresses the functional human glucagon receptor, conferring human relevance to the activity observed with glucagon receptor antagonists. This technique does not provide adequate quantitative precision for the comparative ranking of active compounds, but does afford physiological evidence of efficacy in the early development of a chemical series of antagonists. PMID- 16256356 TI - Seasonal variability in 7Be depositional fluxes at Granada, Spain. AB - Measurement of 7Be depositional fluxes at Granada, Spain (37 degrees 10'50''N-3 degrees 35'44''W, altitude 670 m) in the period 1995 through 1998 indicates substantial variations between the four seasons and also between corresponding seasons in different years, ranging from 23.6 to 242 Bq m(-2) per season. A strongly positive correlation with precipitation is shown, which explains about 70% of the variations in the 7Be depositional fluxes over the 16 seasons studied. The depositional 7Be flux is on the average highest in the fall and lowest in the summer. The study shows that precipitation primarily controls the 7Be depositional flux and plays a dominant role in the removal of 7Be from the troposphere. The average annual 7Be depositional flux at Granada amounts to 469+145 Bq m(-2). PMID- 16256357 TI - Dopaminergic properties and function after grafting of attached neural precursor cultures. AB - Generation of dopaminergic (DA) neurons from multipotent embryonic progenitors represents a promising therapeutical strategy for Parkinson's disease (PD). Aim of the present study was the establishment of enhanced cell culture conditions, which optimize the use of midbrain progenitor cells in animal models of PD. In addition, the progenitor cells were characterized during expansion and differentiation according to morphological and electrophysiological criteria and compared to primary tissue. Here, we report that CNS precursors can be expanded in vitro up to 40-fold and afterwards be efficiently differentiated into DA neurons. After 4-5 days under differentiation conditions, more than 70% of the neurons were TH+, equivalent to 30% of the total cell population. Calcium imaging revealed the presence of calcium-permeable AMPA receptors in the differentiated precursors which are capable to contribute to many developmental processes. The overall survival rate, degree of reinnervation and the behavioral performance after transplantation of 4 days in-vitro-differentiated cells were similar to results after direct grafting of E14 ventral mesencephalic cells, whereas after shorter or longer differentiation periods, respectively, less effects were achieved. Compared to the amount of in-vitro-generated DA neurons, the survival rate was only 0.8%, indicating that these cells are very vulnerable. Our results suggest that expanded and differentiated DA precursors from attached cultures can survive microtransplantation and integrate within the striatum in terms of behavioral recovery. However, there is only a short time window during in vitro differentiation, in which enough cells are already differentiated towards a DA phenotype and simultaneously not too mature for implantation. However, additional factors and/or genetical manipulation of these expanded progenitors will be required to increase their in vivo survival in order to improve both the ethical and the technical outlook for the use of fetal tissue in clinical transplantation. PMID- 16256358 TI - Antiepileptic drug-resistant rats differ from drug-responsive rats in hippocampal neurodegeneration and GABA(A) receptor ligand binding in a model of temporal lobe epilepsy. AB - The disabling seizures associated with mesial temporal lobe epilepsy (TLE) are often resistant to antiepileptic drugs (AEDs). The biological basis of this refractoriness is unknown but may include alterations in AED targets in the epileptogenic brain tissue, reduced AED penetration to the seizure focus, and neuropathological brain alterations such as hippocampal sclerosis typically found in patients with refractory TLE. In the present study, we used a rat model of TLE to examine whether AED responders differ from non-responders in their structural alterations and GABA(A) receptor characteristics in the hippocampal formation. In this model, spontaneous recurrent seizures develop after a status epilepticus induced by prolonged electrical stimulation of the basolateral amygdala. The frequency of these seizures was recorded by continuous video/EEG monitoring before, during, and after daily treatment with phenobarbital, which was given at maximum tolerated doses for 2 weeks. Based on their individual response to phenobarbital, rats were grouped into responders and non-responders. The severity or duration of the initial brain insult (the status epilepticus) did not differ between responders and non-responders, indicating that the difference between the two subgroups is genetically determined. Subsequent histological examination showed a significant loss of neurons in the CA1, CA3c/CA4, and dentate hilus of non-responders, whereas responders did not differ in this respect from non epileptic controls. The morphological alterations in the non-responders were associated with striking alterations in autoradiographic imaging of diazepam sensitive and diazepam-insensitive GABA(A) receptor binding in the dentate gyrus with a significant shift to enhanced diazepam-insensitive binding. The present data indicate that neurodegeneration and associated GABA(A) receptor changes in the dentate gyrus are critically involved in the mechanisms underlying refractoriness of seizures in TLE. PMID- 16256359 TI - Graft placement and uneven pattern of reinnervation in the striatum is important for development of graft-induced dyskinesia. AB - In two recent double-blind clinical trials of fetal ventral mesencephalic cell transplants into the striatum in patients with Parkinson's disease (PD), a significant proportion of the grafted patients developed dyskinetic side effects, which were not seen in the sham operated patients. Comparison between dyskinetic and non-dyskinetic grafted patients in one of the trials suggested that an uneven pattern of striatal reinnervation might be the leading cause of the dyskinesias. Here, we studied the importance of graft placement for the development of dyskinesias in parkinsonian rats. Abnormal involuntary movements resembling peak dose dyskinesias seen in PD patients were induced by daily injections of L-DOPA for 6 weeks. The dyskinetic animals received about 130.000 fetal ventral mesencephalic cells as single grafts placement in the rostral or the caudal aspect of the head of striatum. The results show that grafts placed in the caudal, but not the rostral, part are effective in reducing the L-DOPA-induced limb and orolingual dyskinesia, predominantly seen as hyperkinesia. The same grafts, however, also induced a new type of dyskinetic behavior after activation with amphetamine, which were not seen in non-grafted lesion controls. The severity of these abnormal involuntary movements was significantly correlated with a higher graft-derived dopaminergic reinnervation in the caudal aspect of the head of striatum relative to the rostral part. The results indicate that graft-induced dyskinesias in PD patients may be linked to single, small graft deposits that provide an uneven, patchy reinnervation of the putamen. PMID- 16256360 TI - Mandibular distraction osteogenesis in hemifacial microsomia: long-term follow up. AB - AIM: Prospective, longitudinal, clinical long-term follow-up study of a homogeneous sample of children affected by hemifacial microsomia and treated by mandibular distraction osteogenesis. MATERIAL: Eight patients affected by types I and II hemifacial microsomia were operated on at an average age of 5.6 years with an average follow-up of 5.8 years. METHODS: Vertical changes were measured on postero-anterior cephalometric and panoramic radiographs taken sequentially. RESULTS: Angular changes of the infraorbital and nasal floor planes were not significant, showing that distraction osteogenesis starting after 5 years of age did not influence the maxillary skeletal base. Occlusal (plane) cant was reduced by 7 degrees on average following distraction osteogenesis, showing good dentoalveolar plasticity. Mandibular vertical changes showed a gradual return of the asymmetry, with growth in all patients (the ratio between affected and non affected rami returned by 77% of the correction obtained by means of distraction 5 years postoperatively). CONCLUSION: Although aesthetic and psychological advantages of distraction osteogenesis are well accepted it should only be applied after careful patient selection and honest explanation of the long-term recurrence by genetically determined craniofacial growth patterns. PMID- 16256361 TI - Cell-to-cell contact is critical for the survival of hematopoietic progenitor cells on osteoblasts. AB - Osteoblasts constitute part of the stromal cell support system in marrow for hematopoiesis, however little is known as to how they interact with hematopoietic stem cells (HSCs). In vitro studies have demonstrated that the survival of HSCs in co-culture with osteoblasts requires intimate cell-to-cell contact. This suggests that the osteoblast-derived factor(s) that supports stem cell activities are produced in very small quantities, are rapidly turned over, may be membrane anchored and/or require the engagement of cell-cell adhesion molecules that are yet to be determined. In the present report we found that the survival of hematopoietic progenitor cells on osteoblasts is dependent upon the engagement of VLA-4 (alpha4beta1) and VLA-5 (alpha5beta1) receptors using function blocking antibodies. Cell-to-cell contact is required to support progenitor activity, but can be replaced if receptor-ligand engagement of the VLA-4 and LFA-1 complexes is provided through the use of recombinant ligands (fibronectin, ICAM-1, VCAM-1). Moreover, once these receptors were engaged, conditioned medium derived from HSCs grown on osteoblast ligands supported significantly greater hematopoietic progenitors in vitro than did osteoblast-conditioned or HSC-conditioned medium alone. While the molecules present in the co-cultured medium remain to be identified, the data suggest that hematopoietic cells cooperate with osteoblasts to assemble the various marrow microenvironments by directing the synthesis of osteoblast-derived cytokines to improve HSC survival. PMID- 16256362 TI - Prion protein potentiates acetylcholine release at the neuromuscular junction. AB - Cellular prion protein (PrP(c)), the normal isoform of the pathogenic peptide (PrP(sc)) responsible of the transmissible spongiform encephalopaties (TSEs), is present in many neural tissues, including neuromuscular junctions (NMJ). To analyze if this protein could influence the synaptic transmission, we performed an electrophysiological approach to study the effect of cellular prion protein on a mammalian neuromuscular junction. The loose patch clamp (LPC) technique enables the study of the whole preparation including the pre- and the post-synaptic domains. In a mouse phrenic-diaphragm preparation, nanomolar concentrations of cellular prion protein were able to induce a very striking potentiation of the acetylcholine (ACh) release. The effect was mainly pre-synaptic with an increase of the amplitude of the miniature end-plate currents, probably calcium dependent. Moreover, an apparent facilitation of the synaptic transmission was noted. The results clearly indicate that cellular prion protein may play a key role in the function of the neuromuscular junction. PMID- 16256363 TI - Real-time measurement of signaling and motility during T cell development in the thymus. AB - Intracellular signals arising from interactions of immature thymocytes with distinct populations of stromal cells in the thymus are central to T cell development. The characteristics of these signals and the mechanisms underlying thymocyte migration between stromal cell compartments have been difficult to identify from static measurements of fixed tissue. Recent advances in two-photon microscopy and the development of three-dimensional models for real-time studies of T cell development have shed light on how single cells navigate the thymus. These studies reveal crosstalk between thymocyte signaling and motility that may integrate the search for potentially rare self-antigens with the requirement for sustained signaling in T cell maturation. PMID- 16256364 TI - Altered dopaminergic innervation and amphetamine response in adult Otx2 conditional mutant mice. AB - Here, we have investigated the neurological consequences of restricted inactivation of Otx2 in adult En1(cre/+); Otx2(flox/flox) mice. In agreement with the crucial role of Otx2 in midbrain patterning, the mutants had a substantial reduction in tyrosine hydroxylase containing neurons. Although the reduction in the number of DAergic neurons was comparable between the SNc and the VTA, we found an unexpected selectivity in the deinnervation of the terminal fields affecting preferentially the ventral striatum and the olfactory tubercle. Interestingly, the mutants showed no abnormalities in exploratory activity or motor coordination. However, the absence of normal DA tone generated significant alterations in DA D1-receptor signalling as indicated by increased mutant striatal levels of phosphorylated DARPP-32 and by an altered motor response to amphetamine. Therefore, we suggest that the En1(cre/+); Otx2(flox/flox) mutant mouse model represents a genetic tool for investigating molecular and behavioural consequences of developmental neuronal dysfunction in the DAergic system. PMID- 16256365 TI - Extracellular production of Streptomyces lividans acetyl xylan esterase A in Escherichia coli for rapid detection of activity. AB - Acetyl xylan esterase A (AxeA) from Streptomyces lividans belongs to a large family of industrially relevant polysaccharide esterases. AxeA and its truncated form containing only the catalytically competent domain, AxeA(tr), catalyze both the deacetylation of xylan and the N-deacetylation of chitosan. This broad substrate specificity lends additional interest to their characterization and production. Here, we report three systems for extracellular production of AxeA(tr): secretion from the native host S. lividans with the native signal peptide, extracellular production in Escherichia coli with the native signal peptide, and in E. coli with the OmpA signal peptide. Over five to seven days of a shake flask culture, the native host S. lividans with the native signal peptide secreted AxeA(tr) into the extracellular medium in high yield (388 mg/L) with specific activity of 19 U/mg corresponding to a total of 7000 U/L. Over one day of shake flask culture, E. coli with the native secretion signal peptide produced 84-fold less in the extracellular medium (4.6 mg/L), but the specific activity was higher (100 U/mg) corresponding to a total of 460 U/L. A similar E. coli culture using the OmpA signal peptide, produced 10mg/L with a specific activity of 68 U/mg, corresponding to a total of 680 U/L. In 96-well microtiter plates, extracellular production with E. coli gave approximately 30 and approximately 86 microg/mL in S. lividans. Expression in S. lividans with the native signal peptide is best for high level production, while expression in E. coli using the OmpA secretion signal peptide is best for high-throughput expression and screening of variants in microtiter plate format. PMID- 16256366 TI - Production of active recombinant mitogen-activated protein kinases through transient transfection of 293T cells. AB - Mitogen-activated protein (MAP) kinases are a family of serine/threonine protein kinases that play an important role in a myriad of cellular processes, including cell proliferation, differentiation, and apoptosis. Abnormal activation of MAP kinases has been shown to participate in a variety of human diseases which include cancer, septic shock, rheumatoid arthritis, diabetes, and cardiovascular diseases. Active MAP kinase enzymes are not only valuable for basic biomedical research but are also critical for the development of pharmacological inhibitors as therapeutic drugs in the treatment of relevant human diseases. MAP kinases produced in a bacterial system are poorly active due to a lack of proper phosphorylation at their characteristic threonine and tyrosine residues. To overcome these limitations, we have developed a mammalian expression system for high level expression and one-step purification of enzymatically MAP kinases. We cloned JNK1, p38, and p38-regulated MAP kinase-activated protein kinase-2 into the mammalian expression vector pEBG, and expressed these protein kinases as glutathione S-transferase fusion proteins in human embryonic kidney 293T cells through transient transfection. The protein kinases were activated in vivo through treating the transfected cells with sodium arsenite and affinity-purified using glutathione-Sepharose beads. The enzymatic activities of these protein kinases were demonstrated by Western blot analysis and in vitro kinase assays. Our results indicate that this system is an extremely powerful tool for generating valuable reagents, and could be very valuable for proteomic studies. PMID- 16256367 TI - An efficient method to express and refold a truncated human procaspase-9: a caspase with activity toward Glu-X bonds. AB - A truncated form of human procaspase-9 missing the first 111 amino acids, and a variety of mutants derived therefrom, have been expressed in Escherichia coli inclusion bodies. Upon refolding to active enzymes, Delta(1-111) procaspase-9 and mutants were recovered at purity greater than 95% and with a final yield of 20-35 mg/L cell culture. Our active procaspase-9 retains its pro-segment, while undergoing major auto processing at Asp315 and a minor (20%) cleavage at Glu306. This unusual cleavage at a Glu-X bond also took place in the D315E mutant, and we describe herein the inhibitor Z-VAE-fmk that shows enhanced inactivation of procaspase-9 over caspases-3. The bond at Asp330, not processed by procaspase-9, is cleaved by caspase-3 and the resulting procaspase-9 variant, missing the 316 330 bridge, is six times as active as the non-mutated Delta(1-111) proenzyme. A deletion mutant lacking residues 316-330 underwent auto activation by cleavage at Asp315-Ala331 bond. Moreover, substitution of Glu306 by an Asp residue in this mutant led to rapid removal of the peptide spanning Ser307 to Asp330, and resulted in an enzyme that was 7.6 times as active as the non-mutated Delta(1 111) procaspase-9. Finally, replacing both Asp315 and Glu306 with Ala generated a procaspase-9 mutant incapable of auto processing. This single chain procaspase-9 was fully as active as the non-mutated Delta(1-111) enzyme processed at Asp315 or Glu306. Our demonstration that unprocessed procaspase-9 mutants are active as proteases with caspase-type specificity suggests that the role of procaspase-9 in cascade activation of executioner caspases might, in some circumstances, be carried out alone and without association of the apoptosome. PMID- 16256368 TI - Expression, purification, and characterization of a new heterotetramer structure of leucyl-tRNA synthetase from Aquifex aeolicus in Escherichia coli. AB - Aminoacyl-tRNA synthetases are key players in the interpretation of the genetic code. They constitute a textbook example of multi-domain proteins including insertion and terminal functional modules appended to one of the two class specific active site domains. The non-catalytic domains usually have distinct roles in the aminoacylation reaction. Aquifex aeolicus leucyl-tRNA synthetase (LeuRS) is composed of a separated catalytic site and tRNA anticodon-binding site, which would represent one of the closest relics of the primordial aminoacyl tRNA synthetase. Moreover, the essential catalytic site residues are split into the two different subunits. In all other class-I aminoacyl-tRNA synthetases, those two functional polypeptides are nowadays fused into a single protein chain. In this work, we report the isolation and the characterization, in Escherichia coli, of a novel oligomeric form (alphabeta)2 for A. aeolicus LeuRS, which is present in addition to the alphabeta heterodimer. A. aeolicus (alphabeta)2 LeuRS has been characterized by biochemical and biophysical methods. Native gel electrophoresis, mass spectrometry, analytical ultracentrifugation, and kinetic analysis confirmed that the (alphabeta)2 enzyme was a stable and active entity. By mass spectrometry we confirmed that the heterodimer alphabeta can bind one tRNALeu molecule whereas the heterotetramer (alphabeta)2 can bind two tRNALeu molecules. Active site titration and aminoacylation assays showed that two functional active sites are found per heterotetramer, suggesting that this molecular species might exist and be active in vivo. All those data suggest that the existence of the heterotetramer is certainly not an artifact of overexpression in E. coli. PMID- 16256369 TI - Cell-free synthesis and in situ isolation of recombinant proteins. AB - We present a method for rapid expression and isolation of recombinant proteins. Cell-free protein synthesis in the presence of affinity beads enables in situ isolation of translation products, which simplifies the procedures for the preparation of purified protein samples. In the present study, we have made an attempt to carry out in situ isolation of histidine-tagged proteins by using Ni NTA magnetic agarose beads. The presence of Ni-NTA beads gave no drastic effects on the efficiency of protein synthesis and successfully captured the synthesized proteins. Purified proteins were obtained after subsequent washing and elution steps. In particular, most of the endogenous bead-binding proteins were removed by pre-treating S30 extract with affinity beads and the purity of the target proteins was enhanced up to 95%. The methods described here will provide a basis for fast and convenient preparation of purified proteins from multiple genetic sequences. PMID- 16256370 TI - Programmed cell death progressively models the development of anther sporophytic tissues from the tapetum and is triggered in pollen grains during maturation. AB - To characterize the spatial and temporal occurrence of programmed cell death (PCD) in Lilium anther tissues, we used both microscopical and molecular markers of apoptosis for developmental stages from meiosis to pollen release. The first hallmarks of PCD include cell condensation and shrinkage of the cytoplasm, separation of chromatin into delineated masses, and DNA fragmentation in the tapetum as early as the premeiosis stage. PCD then extended to other anther sporophytic tissues, leading to anther dehiscence. Although the PCD clearly affected the endothecium and the epidermis, these two cell layers remained alive until anther dehiscence. In pollen, no sign of PCD was found until pollen mitosis I, after what apoptotic features developed progressively in the vegetative cell. In addition, DNA ladders were detected in all sporophytic tissues and cell types throughout pollen development, whereas in the male gametophyte DNA ladders were only detected during pollen maturation. Our data suggest that PCD is a progressive and active process affecting all the anther tissues, first being triggered in the tapetum. PMID- 16256371 TI - Meta-perception for pathological personality traits: do we know when others think that we are difficult? AB - The self allows us to reflect on our own behavior and to imagine what others think of us. Clinical experience suggests that these abilities may be impaired in people with personality disorders. They do not recognize the impact that their behavior has on others, and they have difficulty understanding how they are seen by others. We collected information regarding pathological personality traits- using both self and peer report measures--from groups of people who knew each other well (at the end of basic military training). In previous papers, we have reported that agreement between self-report and peer-report is only modest. In this paper, we address the question: Do people know that others disagree with their own perceptions of themselves? We found that expected peer scores predicted variability in peer report over and above self-report for all 10 diagnostic traits. People do have some incremental knowledge of how they are viewed by others, but they do not tell you about it unless you ask them to do so; the knowledge is not reflected in ordinary self-report data. Among participants who expect their peers to describe them as narcissistic, those who agree with this assessment are viewed as being less narcissistic by their peers than those who deny being narcissistic. It therefore appears that insight into how one is viewed by others can moderate negative impressions fostered by PD traits. PMID- 16256372 TI - Being a self: considerations from functional imaging. AB - Having a self is associated with important advantages for an organism. These advantages have been suggested to include mechanisms supporting elaborate capacities for planning, decision-making, and behavioral control. Acknowledging such functionality offers possibilities for obtaining traction on investigation of neural correlates of self-hood. A method that has potential for investigating some of the brain-based properties of self arising in behavioral contexts varying in requirements for such behavioral guidance and control is functional brain imaging. Data obtained with this method are beginning to converge on a set of brain areas that appear to play a significant role in permitting conscious access to representational content having reference to self as an embodied and independent experiencer and agent. These areas have been identified in a variety of imaging contexts ranging from passive state conditions in which they appear to manifest ongoing activity associated with spontaneous and typically 'self related' cognition, to tasks targeting explicitly experienced properties of self, to demanding task conditions where activity within them is attenuated in apparent redirection of cognitive resources in the service of task guidance and control. In this paper, these data will be reviewed and a hypothesis presented regarding a significant role for these areas in enabling degrees of self-awareness and participating in the management of such behavioral control. PMID- 16256373 TI - Contrasting single-trial ERPs between experimental manipulations: improving differentiability by blind source separation. AB - Contrasting event-related potentials (ERPs) generated under different experimental conditions and inferring differential brain responses is widely practiced in cognitive neuroscience research. Traditionally, these contrasts and subsequent inferences have proceeded directly from ERPs measured at the scalp. For certain tasks, it is not unusual that ERPs from a subset of channels are given particular emphasis in data analysis, such as the channels displaying the maximum peak amplitude in regions of interest ("best sensors") or channels showing the largest averaged ERP waveform differences. With the aid of a blind source separation (BSS) algorithm, second-order blind identification (SOBI), which has been recently validated for its ability to recover correlated neuronal sources, we show that single-trial ERPs from previously validated neuronal sources were more distinguishable among different experimental manipulations than the single-trial ERPs measured at the comparable "best sensors". This suggests that by using validated SOBI-recovered neuronal sources, ERP researchers can improve the ability to detect differences in neuronal responses induced by experimental manipulations. Critically, our observations were made at the level of single trials, as opposed to the averaged ERP. Therefore, our conclusions are particularly relevant to phenomena involving trial-to-trial changes in brain activation, for example, rapid induction of brain plasticity and perceptual learning, as well as to the development of brain-computer interfaces. Similar advantages would also apply to analogous situations with magnetoencephalography (MEG). PMID- 16256374 TI - Receptive field size-dependent attention effects in simultaneously presented stimulus displays. AB - Neurophysiological studies in monkeys show that multiple stimuli presented within the receptive field of a neuron are not processed independently but rather act in a mutually suppressive way. Recently, such suppressive interactions have also been reported in human neuroimaging studies. This is seen as evidence that stimuli compete for neural representation. According to the 'biased competition' approach, attention can bias this competition in favor of the attended stimulus, relieving it of the suppressive influences of the distracters. In this paper, we report data that support these findings. Specifically, the effect of attention on stimuli with different spatial separations was investigated more thoroughly. The biased competition approach would predict that, for a given spatial separation and eccentricity, the difference between attended and unattended displays depends on the receptive field size of an area. In a blocked fMRI experiment, participants viewed four simultaneously presented, colorful pictures under different attention conditions (attended and unattended). Stimuli were separated either 2 degrees , 4 degrees or 7 degrees . In line with previous experiments, we found that the effect of attention correlated with the estimated receptive field size of an area. In areas V1, V2 and VP, where estimated receptive field sizes are small, no significant attention effects were found in any of the spatial separation conditions. In V4, there was a significant difference between attended and unattended conditions for the 2 degrees and 4 degrees spatial separations, but not for 7 degrees . Finally in area TEO, significant differences between attended and unattended conditions were observed for all spatial separations. PMID- 16256375 TI - Tracking the subprocesses of decision-based action in the human frontal lobes. AB - Situationally adaptive behavior relies on the identification of relevant target stimuli, the evaluation of these with respect to the current context and the selection of an appropriate action. We used functional magnetic resonance imaging (fMRI) to disentangle the neural networks underlying these processes within a single task. Our results show that activation of mid-ventrolateral prefrontal cortex (PFC) reflects the perceived presence of a target stimulus regardless of context, whereas context-appropriate evaluation is subserved by mid-dorsolateral PFC. Enhancing demands on response selection by means of response conflict activated a network of regions, all of which are directly connected to motor areas. On the midline, rostral anterior paracingulate cortex was found to link target detection and response selection by monitoring for the presence of behaviorally significant conditions. In summary, we provide new evidence for process-specific functional dissociations in the frontal lobes. In target centered processing, target detection in the VLPFC is separable from contextual evaluation in the DLPFC. Response-centered processing in motor-associated regions occurs partly in parallel to these processes, which may enhance behavioral efficiency, but it may also lead to reaction time increases when an irrelevant response tendency is elicited. PMID- 16256376 TI - Cortical abnormalities in bipolar disorder investigated with MRI and voxel-based morphometry. AB - Bipolar disorder (BD) has been associated with abnormalities of brain structure. Specifically, in vivo volumetric MRI and/or post mortem studies of BD have reported abnormalities of gray matter (GM) volume in the medial prefrontal cortex (PFC), amygdala, hippocampal subiculum and ventral striatum. These structures share anatomical connections with each other and form part of a "visceromotor" network modulating emotional behavior. Areas of the lateral orbital, superior temporal and posterior cingulate cortices project to this network, but morphometric abnormalities in these areas have not been established in BD. The current study assessed tissue volumes within these areas in BD using MRI and voxel-based morphometry (VBM). MRI images were obtained from 36 BD subjects and 65 healthy controls. To account for possible neurotrophic and neuroprotective effects of psychotropic medications, BD subjects were divided into medicated and unmedicated groups. Images were segmented into tissue compartments, which were examined on a voxel-wise basis to determine the location and extent of morphometric changes. The GM was reduced in the posterior cingulate/retrosplenial cortex and superior temporal gyrus of unmedicated BD subjects relative to medicated BD subjects and in the lateral orbital cortex of medicated BD subjects relative to controls. White matter (WM) was increased in the orbital and posterior cingulate cortices, which most likely reflected alterations in gyral morphology resulting from the reductions in the associated GM. The morphometric abnormalities in the posterior cingulate, superior temporal and lateral orbital cortices in BD support the hypothesis that the extended network of neuroanatomical structures subserving visceromotor regulation contains structural alterations in BD. Additionally, localization of morphometric abnormalities to areas known to exhibit increased metabolism in depression supports the hypothesis that repeated stress and elevated glucocorticoid secretion may result in neuroplastic changes in BD. PMID- 16256377 TI - An fMRI investigation of short-term source memory in young and older adults. AB - Using functional magnetic resonance imaging (fMRI) and a working memory procedure, we compared source memory judgments (format and location) with old-new judgments in young and older adults. Consistent with previous fMRI findings, for young adults, an area of left dorsolateral prefrontal cortex showed greater activity during format than old-new judgments made immediately, as well as those made after a brief, filled delay. In contrast, for older adults, activity in this area was not greater during format than old-new judgments at either retention interval. These data provide additional evidence that left lateral prefrontal cortex is important in monitoring specific source information and new evidence that older adults' source memory deficits may be related, in part, to reduced function of this brain area. PMID- 16256378 TI - Non-invasive methods and stimuli for evaluating the skin's microcirculation. AB - Vessels in the skin are arranged into superficial and deep horizontal plexuses and they are involved in thermoregulation, oxygen and nutritional support. The skin has a large number of functions and broad appeal spanning basic mechanistic and clinical research. Indeed, the skin can be used as a marker of normal and impaired vascular control and, owing to its accessibility and frequent involvement, is easy to investigate non-invasively. A large number of non invasive methods are available for investigating the skin, ranging from those that permit the visualisation of microvessels, to those that monitor blood flow or one of its derivatives (e.g., skin temperature and transcutaneous oxygen). Such methods can be combined with non-invasive, dynamic stimuli (e.g., the use of cold or warm stimuli, activation of the peripheral nervous system or local neuronal systems, and the topical application of vasoactive drugs) and this potentially enables the differentiation of underlying disorders (e.g., primary from secondary Raynaud's phenomenon) and also to quantify changes over time or following intervention. The present article outlines the non-invasive methods and dynamic tests that can be used to investigate the microcirculation of the skin. PMID- 16256379 TI - The significance of seizures and other predictive factors during the acute illness for the long-term outcome after bacterial meningitis. AB - BACKGROUND: Seizures are important neurological complications of bacterial meningitis, but no information about its epidemiology and the outcomes of seizures after community-acquired bacterial meningitis (CABM) in an adult population have been reported. AIMS: To determine the frequency, clinical relevance, subtypes of seizures during the acute phase of bacterial meningitis, and the long-term outcomes of seizure complicating adult CABM. METHODS: In this 12-year retrospective study, 117 adult patients were identified with culture proven CABM. A comparison was made between the clinical data of the patients with and without seizures during hospitalization. RESULTS: Thirty-one patients had seizures during CABM, accounting for 27% (31/117) of the episodes. The time interval between the onset of bacterial meningitis and the seizures was 1-21 days (mean, 4 days). Furthermore, 80% (25/31) of the episodes occurred within 24 h of presentation. Ten patients who had seizures progressed to status epilepticus. At follow-up after completing treatment, 10 patients completely recovered and were seizure-free, 19 died of meningitis during the acute stage and the other two progressed to chronic epilepsy. CONCLUSION: A log-rank test demonstrated that the long-term outcome of adult CABM with acute seizures produced worse outcomes than for those who had no seizures, though no difference was noted between focal and generalized seizures. None of our patients without seizures in the acute phase of bacterial meningitis developed late seizures during the follow-up periods. Poor outcome in this study may attribute to neurological complications such as seizure, hydrocephalus, infection itself, or a combination of complications. PMID- 16256380 TI - Long-term continuous, but not daily, environmental enrichment reduces spatial memory decline in aged male mice. AB - Although environmental enrichment improves spatial memory and alters synaptic plasticity in aged rodents, it is unclear whether all types of enrichment treatments yield similar benefits. The present study examined the effects in aged male mice of three types of enrichment on spatial memory in Morris water maze and radial arm maze tasks, and on levels of the presynaptic protein synaptophysin in several brain regions. Non-enriched young and aged males were compared with males exposed to one of the following treatments for 10 weeks: 5 min of daily handling, 3 h of daily basic enrichment, or 24 h of continuous complex enrichment. Young controls outperformed aged controls in both tasks. Neither daily handling nor daily enrichment affected spatial memory or synaptophysin levels. In contrast, continuous enrichment significantly reduced age-related spatial memory decline in both tasks, such that this group was statistically indistinguishable from young controls in most measures of performance. Continuously enriched mice were also significantly better than other aged mice in several spatial memory measures. Despite these improvements, synaptophysin levels in the continuous enrichment group were significantly lower than those of young and aged controls in the frontoparietal cortex, hippocampus, and striatum, suggesting a negative relationship between synaptophysin levels and spatial memory in aged males. These data demonstrate that different types of enrichment in aged male mice have disparate effects on spatial memory, and that the relationship between enrichment induced changes in synaptophysin levels and spatial memory in aged males differs from that we have previously reported in aged female mice. PMID- 16256381 TI - Nitric oxide involvement in pancreatic beta cell apoptosis by glibenclamide. AB - Glibenclamide as a second-generation compound of sulfonylurea has widely been used in the treatment of type 2 diabetes patients. It has been shown that it induces apoptosis in beta cells, which is partially mediated by Ca(2+) influx. Here, we investigated the role of nitric oxide (NO) and nitric oxide synthase (NOS) isoforms on glibenclamide-induced apoptosis in rat insulinoma cells. Our results showed that glibenclamide induces NO generation (measured as nitrite) that is accompanied with decrease of cell viability in a defined concentration of glibenclamide. The effects of glibenclamide on cell viability were partially inhibited after treatment with N(G)-nitro-L-arginine methyl ester (L-NAME), inhibitor more selective for constitutive nitric oxide synthase, and in the presence of D600--a blocker of voltage-gated L-type Ca(2+) channels inhibited Ca(2+) influx into beta cells, whereas aminoguanidine (AG), a preferential inhibitor of inducible NOS, was significantly less effective. Analysis of DNA fragmentation by electrophoresis and staining with Hoechest 33342 and propidium iodide showed that L-NAME, but not AG, prevented DNA fragmentation and decreased the number of cells with condensed and fragmented nuclei. It revealed that the effects of glibenclamide on apoptosis were partially inhibited by treatment with L-NAME. In conclusion, we have shown that NO production in glibenclamide treated cells may be involved in the induction of apoptotic cell death in pure beta cell line and it may be due to Ca(2+) dependent activation of constitutive NOS isoforms. PMID- 16256382 TI - Overexpression of inducible nitric oxide synthase and cyclooxygenase-2 in rat zinc-deficient lung: Involvement of a NF-kappaB dependent pathway. AB - Reactive oxygen and nitrogen species have been implicated in the pathogenesis of pulmonary diseases. The goal of this study was to measure the response of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 enzymes (COX-2) in lung with moderate zinc deficiency. Adult male Wistar rats were divided into two groups receiving (1) a zinc-deficient diet (ZD) or (2) a zinc-adequate control diet. After 2 months of treatment, the zinc-deficient group showed a significant pulmonary edema. This was associated to a reduction of protein thiols and to a significant increase of metallothionein and glutathione disulfide levels. In addition, a higher serum and lung NO production in ZD group was positively related to the higher activity and expression of iNOS and COX-2 found in lungs. Western blot analysis revealed increased IkappaBalpha degradation, an indicator of NF-kappaB activation in ZD lungs. Anatomopathologic analysis of ZD lungs showed an increase of connective tissue fibers with an influx of polymorphonuclear cells. These cells and type II cells from the alveoli showed specific immunohistochemical signals for iNOS. The conclusion is that, during the development of zinc-deficiency, iNOS activity increases in lung and contributes to lung injury. Zinc deficiency implications must be taken into account to design therapies and public health interventions involving targeted zinc supplementation for high-risk subjects or certain diseases, such as asthma. PMID- 16256383 TI - Three dimensional magnetic resonance imaging by magnetic resonance force microscopy with a sharp magnetic needle. AB - An electropolished magnetic needle made of Nd(2)Fe(14)B permanent magnet was used for obtaining better spatial resolution than that achieved in our previous work. We observed the magnetic field gradient |G(Z)|=80.0G/microm and the field strength B=1250G at Z approximately 8.8 microm from the top of the needle. The use of this needle for three dimensional magnetic resonance force microscopy at room temperature allowed us to achieve the voxel resolution to be 0.6 microm x 0.6 microm x 0.7 microm in the reconstructed image of DPPH phantom. The acquisition time spent for the whole data collection over 64 x 64 x 16 points, including an iterative signal average by six times per point, was about 10 days. PMID- 16256384 TI - The aqueous reference for ESR oximetry. AB - The interaction of molecular oxygen with derivatives of nitroxide EPR spin labels has been investigated using nuclear spin-relaxation spectroscopy in aqueous and nonaqueous solvents. The proton spin-lattice relaxation rate induced by oxygen provides a measure of the local concentration of oxygen, which we find is dependent on solvent. In water, the hydrophobic effect increases the local concentration of oxygen in the nonpolar portions of solute molecules. For nitroxides reduced to the hydroxylamine in aqueous solutions, we find that the local concentration of oxygen is approximately twice that associated with a free diffusion hard sphere limit, while in octane, this effect is absent. These results show that nitroxide based ESR oximetry may suffer a reference concentration shift of order a factor of two if the aqueous nitroxide spectrum or relaxation is used as the reference. PMID- 16256385 TI - The relation between insulin-like growth factor I levels and cognition in healthy elderly: a meta-analysis. AB - OBJECTIVE: Insulin-like growth factor I (IGF-I) levels and cognitive functioning decrease with aging. Several studies report positive correlations between IGF-I levels and cognitive functioning in healthy elderly. However, because of controversial data no definitive conclusions can be drawn concerning the relation between IGF-I and cognition. Therefore, we carried out a meta-analysis on studies that report on the relation between IGF-I and cognition in healthy elderly. DESIGN: We searched the electronic databases for articles about IGF-I and cognition. Studies from 1985 to January 2005 are included. Two reviewers independently extracted data on study design and cognitive outcomes. Thirteen studies on IGF-I and cognition in elderly, with a total number of 1981 subjects, met the inclusion criteria. On the data from these studies meta-analyses were carried out by means of the program Comprehensive Meta-analysis using a random effects model. RESULTS: Pooled effects show that IGF-I levels in healthy elderly have a positive correlation with cognitive functioning, which appeared to be mainly measured with the mini mental state examination (MMSE). The effect size is 0.6, which indicates the presence of a large positive relationship between IGF and cognition in healthy elderly. CONCLUSION: These meta-analyses showed an overall relationship between IGF-I levels and cognitive functioning in healthy elderly. Further studies should be performed to clarify the role of IGF-I substitution in preserving cognitive functions with aging. PMID- 16256386 TI - Phenylketonuria mutations in Northern China. AB - Mutation spectrum of phenylalanine hydroxylase (PAH) gene in patients with phenylketonuria (PKU) in Northern China is described with a discussion on genotype-phenotype correlation. By using PCR/SSCP and DNA sequencing, all exons of PAH gene in the 185 unrelated patients with PKU from Northern China were studied. A total of 70 different mutations, including 42 missense, 12 splice, 7 nonsense, 5 deletion, 3 insertion, and 1 silence/splice mutations, were detected in 349/370 mutant alleles (94.3%). Deletion, insertion, and frameshift mutations were found for the first time in China PKU patients. The mutations R243Q, EX6 96A>G, R111X, Y356X, and R413P were the prevalent mutations with relative frequencies of 22.2, 11.1, 8.7, 6.5, and 6.5%, respectively. Fifteen novel mutations were identified in this study: I38fsX19, IVS4+3G>C, Y154H, R157K, R157I, T200fsX6, Q267H, Q267E, F302fsX39, G346R, S349A, L367L, R400K, IVS12+4A>G, and IVS12+6T>A. Each of them occurs at very low frequency (0.3-1.1%). The mutation spectrum of PKU in Chinese is similar to other Asian populations but significantly different from European populations. Altogether, 70 different mutations are found in 109 genotypes distributed among 185 PKU patients. As shown by the analysis, the predicted residual activity found in the majority of PKU individuals match their in vivo phenotypes, though evidence is also found for both phenotypic inconsistencies among subjects with similar genotypes and discordance between the in vitro and in vivo effects of some mutant alleles. The study enables us to construct a national database in China serving as a valuable tool for genetic counseling and prognostic evaluation of future cases of PKU. PMID- 16256387 TI - Common polymorphisms (single-nucleotide polymorphisms SNP+45 and SNP+276) of the adiponectin gene regulate serum adiponectin concentrations and blood pressure in young Finnish men. AB - Epidemiologic studies have shown that serum level of adiponectin, a circulating protein secreted by adipocytes, predicts the risk of type 2 diabetes and cardiovascular events. Two single-nucleotide polymorphisms (SNPs) at the adiponectin locus (T45G or G276T) of the adiponectin gene (APM1) have been associated with insulin resistance, low serum adiponectin levels, and diabetes. In the present study, the association of these polymorphisms with serum adiponectin level and insulin resistance-associated risk factors was investigated. To this aim, SNP+45 and SNP+276 of APM1 were genotyped in 252 young Finnish men. Serum adiponectin level (p < 0.001) and diastolic blood pressure (p = 0.031) were significantly higher in subjects with the T276T genotype of APM1 compared to those with the G276T or G276G genotypes. Mean diastolic blood pressure among the T276T subjects was 80 mmHg and that in subjects with the G276G and G276T genotypes below 75 mmHg. An interaction between triglycerides, diastolic blood pressure, quantitative insulin sensitivity check index, and SNP276 with regard to serum adiponectin level was found. After adjustment for other covariates, the interaction between triglycerides and SNP276 remained statistically significant (p = 0.009). Among subjects with the T276T genotype, an increase in triglyceride level was associated with a decrease in adiponectin concentration. This result was not observed in other genotype groups. SNP+45 was not significantly related to serum adiponectin concentration, but high-density lipoprotein (HDL) cholesterol tended to be higher in subjects with the T45T genotype (p = 0.051) compared to subjects with the X45G genotype. In conclusion, the T276T genotype of the adiponectin gene was associated with elevated serum adiponectin level and diastolic blood pressure among young Finnish men. PMID- 16256388 TI - The use of yeast mitochondria to study the properties of wild-type and mutant human mitochondrial ornithine transporter. AB - Yeast deletion models have general utility for the study of a variety of inherited metabolic disorders. Mutations in the mitochondrial ornithine transporter result in hyperammonemia, hyperornithinemia, homocitrullinuria syndrome, a disorder of the urea cycle. To study the effects of mutations in a model system that more closely resembles the in vivo environment, we have developed an expression system based on a yeast strain lacking its endogenous ornithine transporter homologue. Wild-type human ornithine transporter and a recurrent mutation, DeltaF 188, were expressed and characterized with this system. The wild-type transporter appeared to insert into yeast mitochondria in the same orientation as in mammalian mitochondria. It showed stereospecificity, strong antiport activity and ornithine transport was competed by citrulline and arginine. The DeltaF 188 mutant was not incorporated into the membrane to the same extent as wild type, but retained significant residual activity and lost stereospecificity. In these isolated mitochondria, samarium chloride was found to be a potent blocker of transport compared to previously reported sulfhydryl-based inhibitors. A low-affinity background transport activity that promoted the exchange of ornithine for either acidic or basic amino acids was observed. This yeast model can readily be extended to the study of protein:protein interactions. In this manner, the use of yeast deletion strains can serve as a general framework to perform metabolic pathway analysis. PMID- 16256390 TI - First characterization of a large deletion of the PDHA 1 gene. AB - Pyruvate dehydrogenase complex (PDC) deficiency is one of the major recognized causes of congenital lactic acidosis. The most common form is due to PDHA 1 gene (Xp22.12) defects. Here, we report the case of a Polynesian girl presenting with delayed neurological development, cortical atrophy, and posterior corpus callosum agenesis. Elevated lactate and pyruvate levels in blood and cerebrospinal fluid suggested PDC deficiency. However, PDC activity was within the normal range in lymphocytes and the direct sequencing of the 11 exons and intron-exon junctions of the PDHA 1 gene did not show any changes. Long-range PCR amplification of the whole gene (16 kb) from blood DNA revealed a heterozygous deletion of approximately 4.2kb. Fine mapping of the deletion breakpoint was achieved using purified long-range PCR products for restriction enzyme analysis and direct sequencing. The deletion removed a 4,227 bp region covering part of intron 5 to part of intron 9 [g.10,145_14,371 del 4,227]. The deletion breakpoint contained a short direct repeat (GTAG), which may be derived either from the upstream or the downstream homologous sequence. The presence of a GAG triplet and inverted repeats in the vicinity of the deletion suggest replication slippage at a polymerase alpha arrest site. This is the first time that a large intragenic deletion of the PDHA 1 gene has been characterized. PMID- 16256389 TI - A known functional polymorphism (Ile120Val) of the human PCMT1 gene and risk of spina bifida. AB - Folate binding protein 1 (Folr1) knockout mice with low maternal folate concentrations have been shown to be excellent animal models for human folate responsive neural tube defects (NTDs). Previous studies using the Folr1 knockout mice revealed that maternal folate supplementation up-regulates the expression of the PCMT1 gene in Folr1 nullizygous neural tube tissue during neural tube closure. PCMT1 encodes the protein repair enzyme l-isoaspartate (d-aspartate) O methyltransferase (PIMT) that converts abnormal d-aspartyl and l-isoaspartyl residues to the normal l-aspartyl form. PIMT is known to protect certain neural cells from Bax-induced apoptosis. Pcmt1-deficient mice present with abnormal AdoMet/AdoHcy homeostasis. We hypothesized that a known functional polymorphism (Ile120Val) in the human PCMT1 gene is associated with an increased risk of folate-responsive human NTDs. A case-control study was conducted to investigate a possible association between this polymorphism and risk of spina bifida. Compared to the Ile/Ile and Ile/Val genotypes, the homozygous Val/Val genotype showed decreased risk for spina bifida (adjusted odds ratio=0.6, 95% confidence interval: 0.4-0.9). Our results showed that the Ile120Val polymorphism of PCMT1 gene is a genetic modifier for the risk of spina bifida. Val/Val genotype was associated with a reduction in risk for spina bifida. PMID- 16256391 TI - Delivery of exogenous tetrahydrobiopterin (BH4) to cells of target organs: role of salvage pathway and uptake of its precursor in effective elevation of tissue BH4. AB - Cells in target organs such as liver do not generally incorporate tetrahydrobiopterin (BH4) in its fully reduced form. Instead, they transiently take up BH4 from the extracellular fluid, instantaneously oxidize it and then expel virtually all of it. However, a small but stable accumulation of BH4 was observed after BH4 administration to the cell cultures. This accumulation was inhibited by methotrexate, an inhibitor of dihydrofolate reductase, a phenomenon that was first suggested based on results of in vitro studies which used established cell lines such as RBL2H3 and PC12. These cells also take up dihydrobiopterin (BH2) and reduce it to enzymically active BH4. Their ability to accumulate usable BH4 upon BH4 administration was attributed to the incorporation of BH2, which in typical experiments was produced by the cells as well as by auto oxidation of BH4. Most cells of the various cell lines so far examined behaved similarly in culture. Our in vivo work with individual mice demonstrated that administration of sepiapterin, BH2, and BH4 was comparably effective in raising BH4 levels in target organs. BH4 accumulation in various tissues after supplementation with BH4, BH2 or sepiapterin was also inhibited by methotrexate, as in the case of our cell culture system. It was concluded that the elevation in BH4 by supplementation was mainly through a "salvage pathway" that included BH2 as the key intermediate in the production of BH4 through the action of dihydrofolate reductase. PMID- 16256392 TI - [Radiation oncology training in France: demography, analysis of motivations of the young specialists, evaluation of the training]. AB - During the 5 past national courses organised by the French society of radiation oncology (SFRO), three different types of survey were performed to analyse demography, motivations and quality of training of the young specialists. During the 5 past years, 50 radiation oncologists were training for the whole country (about 15 per year were graduated). A recent increase the number of young specialists is observed with a total number of 50 in 2000 to 75 in 2005. Nevertheless, the number of young specialists is dramatically insufficient and exposes for the future to an important demographic crisis. Analysis of motivations of choice for radiation oncology confirms the influence of a practical stage of oncology during the second cycle of the medical studies for 60% of the young specialists. Analysis of practical and theoretical training was performed according to the point of view and living experiences of the students. On the other hand, informations from teachers were less complete. Some needs are emphased as: 1) the quality of the follow during the training (importance of the recent implementation of a logbook); 2) importance of theoretical and practical training at the radiotherapy department; 3) help and incentive for research and scientific publication. PMID- 16256393 TI - Intraarticular glucocorticoid injections in rapidly destructive hip osteoarthritis. AB - OBJECTIVE: To determine whether an intraarticular glucocorticoid injection followed, when possible, by weight-bearing elimination using two crutches reduces the need for total hip arthroplasty (THA) in patients with rapidly destructive hip osteoarthritis (RDHOA). METHODS: A longitudinal retrospective study was conducted in patients admitted for RDHOA, defined as loss of more than 50% of the joint space at the narrowest point between two evaluations 1 year apart. A glucocorticoid injection was performed under fluoroscopic guidance. Patients stayed in bed for the next 24 h then used crutches for 4-6 weeks. Follow-up was at least 6 months. The efficacy criterion was absence of THA. RESULTS: Twenty eight patients (22 women) were enrolled between 1993 and 2000. Mean age was 62 years, mean body mass index was 26 kg/m(2), mean Lequesne index was 11, and mean joint space width was 1.3 mm. Narrowing was superolateral in 19 of the 28 patients. Cortivazol was injected in seven patients, betamethasone in four, and triamcinolone hexacetonide in 17. Weight-bearing elimination for at least 4 weeks was achieved in 15 patients. THA was performed in 27 patients, including 20 who underwent the procedure within the year after the glucocorticoid injection. CONCLUSION: Intraarticular glucocorticoid injection with or without elimination of weight-bearing does not reduce the need for THA in patients with RDHOA. PMID- 16256394 TI - Development of cervical interspinous bursitis after prolonged sports trauma: a case report. PMID- 16256395 TI - Use of complementary and alternative therapies by patients with self-reported chronic back pain: a nationwide survey in Canada. AB - OBJECTIVE: Complementary and alternative medicine (CAM) is gaining popularity among patients with chronic back pain. We looked for factors associated with CAM use. METHODS: The data came from the cross-sectional household component of the 1996-1997 National Population Health Survey on the health status and behaviors of Canadians. The sample comprising 66.999 individuals aged 20 years or older represented 21 million adults. Cross tabulations were used to estimate the percentage of CAM use among adults with chronic back pain. Factors independently associated with CAM use during the year before the surveys were identified using multiple logistic regression. RESULTS: CAM use was highest in the subgroup of Canadian adults reporting chronic back pain (39.07%). CAM use was associated with younger age, being married, having a higher level of education, and earning a higher income. Overall, the CAM users reported less pain, greater analgesic use, more depression, and more co-morbidities. In addition to CAM, these patients used conventional medical services. CONCLUSION: Our results show that patients with chronic back pain who use CAM are more active, more involved in social life, and healthier, suggesting better management of their condition. They use CAM in addition to, rather than instead of, conventional care. CAM use in these patients may be ascribable to dissatisfaction with mainstream physicians. PMID- 16256397 TI - Systemic sarcoidosis: the "leopard-man" sign. AB - Systemic sarcoidosis is often diagnosed late. A 37 year-old man from the Antilles was admitted for evaluation of arthralgia, subcutaneous and cutaneous nodules, multiple enlarged lymph nodes, and an epididymal lump. He had been given a diagnosis of epididymal and nodal tuberculosis but had not responded to antitubercular agents. Gallium-67 scintigraphy showed multiple hot spots in the soft tissues, skin, muscles, mediastinum, and lachrymal glands, producing a diffuse dappled pattern. Magnetic resonance imaging of the thighs confirmed the muscular and subcutaneous involvement. Systemic sarcoidosis was diagnosed. Prednisone therapy was promptly effective. We suggest the term "dappled-body sign" or "leopard-man sign" to designate the heretofore undescribed scintigraphic pattern in our patient. PMID- 16256396 TI - Increased serum thrombopoietin concentration in systemic sclerosis associated with myelofibrosis and thrombocytosis. PMID- 16256398 TI - Rheumatoid arthritis in 2005: which patients should receive TNFalpha antagonists and when? A point of view. PMID- 16256399 TI - [Complications of laparoscopic lymphadenectomy in gynaecologic oncology. A series of 1102 procedures in 915 patients. Gynecol Obstet Fertil 2005;33:304-314]. PMID- 16256400 TI - [Hereditary predispositions to gynaecological cancers]. AB - The breast, ovary and endometrial cancers are hereditary in 5 to 10% of the cases. These genetic predisposition syndromes can be classified into two major classes: ovarian cancer and breast cancer predisposition family cases (genes BRCA1 and BRCA2) and family cases of colon cancer, endometrial cancer and ovarian cancer (Lynch syndrome or HNPCC) (genes hMLH1, hMSH2, hMLH6). The estimate of the family and individual risk can contribute in a determining manner to the management of these patients, by the practice of screening or an adapted prevention. Indeed, the risk of cancer of an individual having a positive test for a gene of predisposition to breast cancer (BRCA1, BRCA2) or to the colon cancer (hMLH1, hMSH2, hMLH6) lies between 50 and 70% at the age of 70 years. The indication of a genetic test must be discussed within the framework of an oncogenetic consultation. An individual and family medical management ranging from simple monitoring to prophylactic surgery is proposed to these predisposed people. PMID- 16256401 TI - [Against systematic ultrasound-guided embryo transfer]. PMID- 16256402 TI - [Influence of pregnancy on renal angiomyolipoma]. AB - The aim is to perform a literature search on the role of pregnancy in the rupture of renal angiomyolipoma. Articles published from 1952 to 2004 in the Medline database were searched using the keywords renal angiomylipoma and pregnancy. Pathologies associated with angiomylipoma (lymphangioleiomyomatosis and Bourneville tuberous sclerosis) were taken into account. Seventy-two cases of association of renal angiomyolipoma and pregnancy were found, out of which 58 presented a haemorrhage. In only 26% of the cases, renal angiomyolipoma had been documented prior to pregnancy. Clinical presentation was similar to cases occurring among non pregnant women: abdominal pains (88%), hypotension or shock (33%) and hematuria (24%). Average size of the rupture was 11,7 cm. Rupture does not occur solely with the first pregnancy but occurred equally during the first, second and third pregnancy. Average gestation age upon occurrence of haemorrhage was 27 weeks with a minimum at ten weeks. Therapeutic strategies at the time of shock or hypotension were total nephrectomy in 79% of the cases, 7% polar nephrectomy, 7% embolisation followed by nephrectomy and 7% abstention. A causal role of pregnancy in the atraumatic rupture of angiomyolipoma is not clearly defined. Nevertheless, many arguments (whether it be abdominal mechanical pressure, hormonal or histological ones) suggest that a pregnancy could increase the risk of renal angiomyolipoma rupture. These patients should have a medical follow-up period at closer intervals during their pregnancy and the postpartum phase. PMID- 16256403 TI - [Mammary tuberculosis: a retrospective study of 65 cases]. AB - OBJECTIVES: Remember the clinical, morphologic and histological particularities of mammary tuberculosis. Discuss clinical and radiological diagnostic problems caused by this localization, as far as breast cancer is concerned. PATIENTS AND METHODS: This is a retrospective study concerning 65 women suffering from mammary tuberculosis, diagnosed at Salah Azaiez Institute between January 1980 and December 2001. RESULTS: In this series, mammary tuberculosis represents 0.2% of the declared tubercular localizations and 0.3% of the mammary pathologies treated at the institute. The mean age of our patients was 36 years (19 to 79). Clinical findings were misleading. In 60% of cases the aspect was of a malignant tumor, in 24.6% of cases of a benign nodule and in 16.4% of cases of an abscess. The mammography concluded to a malignant lesion in 49% of cases. The fine needle aspiration achieved at 8 patients was negative in 6 cases and brought back the caseum in the 2 others. The diagnosis has been made, for all our patients, after histological study when we found typical tuberculosis lesions on pieces of tumorectomy, or biopsy. The diagnosis of tuberculosis was established; all patients had an anti-tuberculosis medical treatment in a department of infectious diseases out of the institute. DISCUSSION AND CONCLUSION: Clinical and radiological features of mammary tuberculosis are very confusing and cause a diagnostic problem with breast cancers. In front of symptoms evoking tuberculosis, biopsies must be done to eliminate an eventual cancer. PMID- 16256404 TI - Correction for broadening in Doppler blood flow spectrum estimated using wavelet transform. AB - The conventionally used spectral estimation technique for Doppler blood flow signal analysis is short-time Fourier transform (STFT). But this method requires stationarity of the signal during the window interval. Wavelet transform (WT), which has a flexible time-frequency window, is particularly suitable for nonstationary signals. In recently years, the WT has been used to investigate its advantages and limitations for the analysis of Doppler blood flow signals. In these studies, the estimated spectral width of Doppler blood flow signals using the WT might include significant window and nonstationarity broadening errors. These broadening errors of the time-varying spectrum were clearly undesirable since it would tend to mask the effect of flow disturbance on the spectra width. In this paper, a closed form expression for window and nonstationary root-mean squared (rms) spectral width is given when using the WT to estimate the Doppler blood flow spectrum. The increases in the rms spectral width can be calculated and then the spectral width estimation based on the WT can be corrected. PMID- 16256406 TI - Isolated continuous rhythmic involuntary tongue movements following a pontine infarct. AB - Isolated involuntary tongue movements are rare and poorly understood. The anatomical substrate and pathogenesis underlying involuntary tongue movements remain elusive. We describe a patient who developed isolated continuous rhythmic involuntary tongue movements after pontine infarct without evidence of hypertrophy of inferior olivary nucleus on follow-up magnetic resonance image. We discuss the rhythmic involuntary tongue movements as a prototype of involuntary hyperkinetic movement released by a central pacemaker. PMID- 16256405 TI - The constitutive properties of the brain paraenchyma Part 2. Fractional derivative approach. AB - Fractional models have proven to be very useful for studying viscoelastic materials. We consider the fractional Zener model (also called four-parameter model) to study both the relaxation function and creep compliance. The analytical results are compared with the known experimental results of the human brain tissue to obtain the best fit and brain mechanical parameters. The results are also compared to the non-fractional Zener model and four-parameter Burgers model, indicating that the four-parameter fractional model gives a substantially better fit for the all experimental data. PMID- 16256407 TI - The effects of external cues on ankle control during gait initiation in Parkinson's disease. AB - The present study investigated the effects of external cues on motor control of the ankle joint during gait initiation in patients with Parkinson's disease (PD) and in age-matched healthy subjects. The soleus H-reflexes were recorded during self-generated and cue-triggered gait initiation. The tibialis anterior muscle burst during cue-triggered gait initiation was found to be significantly larger than that during self-generated gait initiation in both groups. External cues significantly increased soleus H-reflex depression during gait initiation in PD patients, although this significant increase was not present in healthy subjects. These findings indicate that external cues affect motor control of the extensor muscle of the ankle joint during gait initiation in PD patients. PMID- 16256408 TI - Parkinsonism with basal ganglia lesions in a patient with uremia: Evidence of vasogenic edema. AB - Parkinsonian syndromes associated with basal ganglia pathology have very rarely been reported in patients with end-stage renal failure. The nature and pathophysiology of the basal ganglia lesion responsible for parkinsonism were unknown. A 48-year-old man who had advanced renal failure developed disturbance of balance and gait and decreased spontaneity. Brain magnetic resonance (MR) imaging disclosed bilateral basal ganglia lesions. By the finding of diffusion weighted image, the apparent diffusion coefficient map, MR angiography, and SPECT, we suggest that the basal ganglia lesions may be the result of vasogenic edema attributable to focal hyperemia secondary to abnormal dilatation of small vessels. PMID- 16256409 TI - The LRRK2 I2012T, G2019S, and I2020T mutations are rare in Taiwanese patients with sporadic Parkinson's disease. PMID- 16256410 TI - Parkinson's disease in diphenyl-exposed workers--a causal association? AB - We report a cluster of five cases of Parkinson's disease (PD) among paper mill workers exposed to a fungicide, diphenyl. The cause of PD is still unknown, but epidemiological studies have indicated an elevated risk of developing PD after exposure to pesticides. The five cases of PD were found in a group of 255 diphenyl-exposed workers, and the number of expected cases in the exposed group was estimated to be 0.9, resulting in a relative risk of 5.6 (95% CI 1.8-13). Exposure to diphenyl may have contributed to this PD cluster, but chance is an alternative explanation. PMID- 16256411 TI - Epidermal fatty acid-binding protein is increased in rat lungs following in vivo treatment with keratinocyte growth factor. AB - Exogenous application of keratinocyte growth factor protects the lung against a variety of injurious stimuli. KGF-treatment leads to pronounced hyperplasia of alveolar epithelial type II cells and to stabilization of surfactant homeostasis after lung injury. Epidermal fatty acid-binding protein is involved in the synthesis of surfactant phospholipids and acts as an antioxidant scavenging reactive lipids. We treated adult rats with recombinant human keratinocyte growth factor (Palifermin) via intratracheal instillation and analyzed the expression of epidermal fatty acid-binding protein mRNA and protein by quantitative RT-PCR, immunoblotting as well as immunohistochemistry. Keratinocyte growth factor treatment in vivo leads to an increased expression of epidermal fatty acid binding protein mRNA and protein in the total lung. Epidermal fatty acid-binding protein mRNA expression per alveolar epithelial type II cell remains constant as shown in isolated type II cells. Epidermal fatty acid-binding protein immunoreactivity is seen in most if not all hyperplastic alveolar epithelial type II cells, and is mainly localized to the cytoplasm. The increase in epidermal fatty acid-binding protein gene expression associated with type II cell hyperplasia might contribute to the molecular mechanisms mediating lung protection by keratinocyte growth factor. PMID- 16256412 TI - Oral malignant neoplasia: a survey of 428 cases in two Zimbabwean hospitals. AB - A review of oral malignant neoplasms biopsies accessioned in the pathology services of Harare Central Hospital and Parirenyatwa Hospital, Harare, Zimbabwe, during the 10year period January 1982 to December 1991 was carried out. Clinical records of 428 patients with histologically diagnosed oral malignant neoplasms by ICD-O type and site (COO.O-CO6.9) were retrieved and analyzed for gender, age, history of tobacco and alcohol usage, neoplastic type and histologic differentiation of squamous cell carcinoma (8000/3). Oral malignant neoplasms constituted 24.8% (n = 428) of the 1723 biopsies accessioned in the department during this period. There was a male to female ratio of 1.9:1, with lesions being most common in the 41-60 year age group. Squamous cell carcinoma, (8070/3) 73.1%, was the predominant oral malignancy most common in the 61-70 year age group with 21.1% on the mandibular gingivae (CO3.1), 20.5% on the tongue (CO1.9-CO2.9), 18.5% on the floor of the mouth (CO4.9) and 10.5% each on the maxillary gingivae (CO3.O) and buccal mucosa (CO6.O). The hard palate (CO5.9) had 9% of the lesions. Lip malignancy (COO-COO.9 and C44.0), inclusive of mucosa of lip, vermilion and skin, constituted 3.3% and minor salivary gland (CO6.9) malignancy (M-8940/3) 4.9%. The remaining oral malignancies, 11%, included Burkitt's lymphoma (M 9687/3) 4.7%; Kaposi's sarcoma (M-9140/3) 3.7%; osteosarcoma (M-9180/3) 2.6% and malignant melanoma (M-8720/3) 1.9%. Fourty five percent of the patients admitted tobacco and alcohol consumption, of whom 95% were males. Oral malignant neoplasms were more common in males than females and occurred in a relatively young age group. Well-differentiated squamous cell carcinoma (M-8070/31) was the most common oral malignant neoplasm and was most frequently found on the mandibular gingivae (CO3.1), tongue (CO1.9-CO2.9) and floor of the mouth (CO4.9). The lip (COO-COO.9) was least affected with a high palatal (CO5.9) involvement. PMID- 16256414 TI - Elective neck treatment versus observation in patients with T1/T2 N0 squamous cell carcinoma of oral tongue. AB - A retrospective analysis of 80 patients treated for T1/T2 N0 squamous cell carcinoma of oral tongue was performed. The patients were divided into two groups according to the management of the neck: those without (n = 34) and those with elective neck treatment (n = 46). The two groups were compared with respect to overall survival (OS), disease specific survival (DSS), and recurrences. The incidence of occult nodal disease and the results of salvage treatment were analysed. There were significantly fewer regional recurrences in patients with elective neck treatment. Statistical differences in OS or DSS between the treatment groups were not found. Of all the patients with locoregional recurrence, 33% were salvaged. The incidences of occult metastasis for T1 and T2 tumours were 24% and 35%, respectively. The elective neck treatment resulted in better regional control. The results of salvage treatment were poor. The risk for occult cervical metastasis is high in patients with early tongue tumours and only carefully selected patients can be left without prophylactic neck treatment. PMID- 16256413 TI - Immunohistochemical expression of p63 in pleomorphic adenomas and carcinomas ex pleomorphic adenomas of salivary glands. AB - Alteration of the p63 protein expression has been associated with several human cancers including salivary gland tumors. We sought to assess the p63 expression in pleomorphic adenomas (PA) and carcinoma ex-pleomorphic adenomas (Ca ex-PA), since this protein has been related to myoepithelial phenotype and the biological behavior of cancer. Ten samples of PA and ten of Ca ex-PA were submitted to immunohistochemistry using a monoclonal anti-p63 antibody, clone 4A4. p63 was regularly expressed in myoepithelial cells' nuclei of the normal glandular parenchyma and in PA, but it was completely absent in five out of the ten samples of Ca ex-PA. (P < 0.05 by Fisher's exact test). Apparently, only those cases depicting some myoepithelial differentiation were positive. Our data suggest that loss of myoepithelial differentiation is important in the evolution of Ca ex-PA, and corroborate the hypothesis that p63 antigen may be a useful marker of myoepithelial cells in salivary glands neoplasms. PMID- 16256415 TI - Toluidine blue uptake in potentially malignant oral lesions in vivo: clinical and histological assessment. AB - To assess the histological features of in vivo toluidine blue (TB) uptake in potentially malignant oral lesions (PML) and to determine whether any were related to clinical Dark versus Pale Royal Blue stain and/or to the malignant/dysplastic nature of the lesions. Frozen sections were used to evaluate TB extra- and intra-epithelial distribution, depth of penetration and nuclear or extra-nuclear uptake. Eighteen lesions were studied. The clinical appearance of a Dark Royal Blue stain was significantly related to the nuclear uptake of the dye. Conversely Pale Royal Blue staining was unrelated to any histological feature. Dark Royal Blue-malignant lesions had more nuclear uptake than benign lesions. The results suggest that Dark Royal Blue staining is the true positive outcome of a TB test and showed that Dark Royal Blue-malignant and Dark Royal Blue-benign lesions have a different histological pattern of uptake. PMID- 16256416 TI - A single nucleotide polymorphism in the matrix metalloproteinase-1 promoter enhances oral squamous cell carcinoma susceptibility in a Chinese population. AB - We genotyped 96 oral squamous cell carcinoma (OSCC) patients for the 1G/2G polymorphism of matrix metalloproteinase-1 (MMP-1) promoter -1607 bp using PCR RFLP. A control population of 120 frequency-matched subjects was also genotyped for the same polymorphism. The detection frequency of 2G allele was significantly higher in OSCC subjects (76%) than in the control group (56.7%). The frequency of 2G allele had a significant difference between the OSCC and controls group (p = 0.00, Odds Ratio, OR = 2.232, 95% CI = 1.477-3.372). The genotype 2G/2G was found in 57.3% of the OSCC, and 34.2% in the controls. The proportion of 2G homozygote (2G/2G) was significantly higher in the OSCC group when compared to controls (p = 0.001, OR = 2.585, 95% CI = 1.487-4.494). OSCC patients were stratified by clinicopathological parameters including gender, smoking, clinical stage and lymph node metastasis, but the only statistically significant association with MMP-1 genotype was with smoking. The results showed that a SNP in the MMP-1 promoter -1607 bp was associated with OSCC susceptibility in a Chinese population. PMID- 16256417 TI - Squamous cell carcinoma of the oral cavity, maxillary antrum and lip in a Zimbabwean population: a descriptive epidemiological study. AB - Squamous cell carcinoma accounts for approximately 90% of oral malignancies. The objective of this study was to document the gender, age, sub-site distribution and histologic differentiation of squamous cell carcinoma of the oral cavity, maxillary antrum and lip in a Zimbabwean population. Hospital records of patients with a histologic diagnosis of squamous cell carcinoma of the oral cavity, maxillary antrum and lip seen at Harare Central Hospital and Parirenyatwa Hospital in Zimbabwe during the period January 1982-December 1991 were reviewed. 20.8% (n = 358/1723) were squamous cell carcinoma of the oral cavity, maxillary antrum and lip. Age ranged from 3 to 70years with a 2:1 male:female ratio. Peak incidence in both sexes were in the 41-50 and 51-60years age groups. Sub-site distribution was mandibular gingiva 18.4%, tongue 17.9%, floor of the mouth 16.2%, maxillary gingiva 9.2%, buccal mucosa 9.2%, maxillary antrum 12.6%, hard palate 7.8%, soft palate 4.8%, lower lip 2.8% and upper lip 1.1%. 64.8% were well differentiated, 24.8% moderately differentiated and 10.4% poorly differentiated. The mandibular gingiva, floor of the mouth and tongue were most commonly affected. Lip squamous cell carcinoma was uncommon. Well-differentiated squamous cell carcinoma was most common in the 41-60 years age group. PMID- 16256418 TI - Archaeal chromatin proteins: different structures but common function? AB - Chromatin proteins promote chromosome flexibility in vivo, maintaining a compact yet decondensed template that permits polymerase accessibility. All Archaea have at least two types of chromatin proteins, and diversity in the chromatin protein population appears to prevent polymerization of a single type of protein. Of the numerous chromatin proteins that have been described in Archaea, only two- histones and Alba homologs--are present in all archaeal phyla. Although their structures and complexes with DNA have no similarities, their functions probably overlap as mutants that lack single chromatin proteins are viable. PMID- 16256419 TI - Unusual pathways and enzymes of central carbohydrate metabolism in Archaea. AB - Sugar-utilizing hyperthermophilic and halophilic Archaea degrade glucose and glucose polymers to acetate or to CO2 using O2, nitrate, sulfur or sulfate as electron acceptors. Comparative analyses of glycolytic pathways in these organisms indicate a variety of differences from the classical Emden-Meyerhof and Entner-Doudoroff pathways that are operative in Bacteria and Eukarya, respectively. The archaeal pathways are characterized by the presence of numerous novel enzymes and enzyme families that catalyze, for example, the phosphorylation of glucose and of fructose 6-phosphate, the isomerization of glucose 6-phosphate, the cleavage of fructose 1,6-bisphosphate, the oxidation of glyceraldehyde 3 phosphate and the conversion of acetyl-CoA to acetate. Recent major advances in deciphering the complexity of archaeal central carbohydrate metabolism were gained by combination of classical biochemical and genomic-based approaches. PMID- 16256420 TI - The direct genetic encoding of pyrrolysine. AB - Pyrrolysine is an amino acid encoded by the amber codon in genes required for methylamine utilization by members of the Methanosarcinaceae. Pyrrolysine and selenocysteine share the distinction of being the only two non-canonical amino acids that have entered natural genetic codes. Recent experiments have shown that encoding of pyrrolysine, unlike that of selenocysteine, also shares an important trait of the original set of twenty amino acids. UAG is translated as pyrrolysine with the participation of a dedicated aminoacyl-tRNA synthetase. Expression of the genes encoding the pyrrolysyl-tRNA synthetase and its cognate tRNA is sufficient to add pyrrolysine to the genetic code of a recombinant organism. Thus, the recruitment of pyrrolysine into the genetic code involved evolution of the first non-canonical aminoacyl-tRNA synthetase and cognate tRNA to be described from nature. PMID- 16256421 TI - Small non-coding RNAs in Archaea. AB - Biochemical and informatics analyses conducted over the past few years have revealed the presence of a plethora of small non-coding RNAs in various species of Archaea. A large proportion of these RNAs contain a common structural motif called the RNA kink turn (K-turn). The best-characterized are the C/D box and the H/ACA box guide small (s)RNAs. Both contain the K-turn fold and require the binding of the L7Ae protein to stabilize the structure of this crucial motif. These sRNAs assemble with L7Ae and several other proteins into complex and dynamic ribonucleoprotein machines that mediate guide-directed ribose methylation or pseudouridylation to specific locations in ribosomal or transfer RNA. Analyses of new archaeal sRNA libraries have identified additional classes of novel sRNAs; many of these contain the RNA K-turn motif and suggest that the RNAs might function as ribonucleoprotein complexes. Some have characteristics of small interfering RNAs or of micro RNAs that have been implicated in the post transcriptional control of gene expression, whereas others appear to be involved in protein translocation or in ribosomal RNA processing and ribosome assembly. A complete understanding of the structure of the K-turn motif and its contribution to various RNA-RNA and RNA-protein interactions will be absolutely essential to fully elucidate the biological organization, activity and function of these novel archaeal ribonucleoprotein machines. PMID- 16256422 TI - Stress response by solute accumulation in archaea. AB - The accumulation of organic solutes is a prerequisite for osmotic adjustment of all organisms. Archaea synthesize unusual solutes such as beta-amino acids, Nepsilon-acetyl-beta-lysine, mannosylglycerate and di-myo-inositol phosphate but, as in other cells, uptake of solutes such as glycine betaine is preferred over de novo synthesis. Study of the molecular basis of osmoadaptation and its regulation in archaea is still in its infancy, but genomics and functional genome analyses combined with classical biochemistry shed light on the processes that confer osmoadaptation in archaea. Most interestingly, some solutes are not only produced in response to salt but also to temperature stress. PMID- 16256423 TI - Archaeal proteasomes and other regulatory proteases. AB - Numerous proteases have been shown to catalyze the precisely-timed and rapid turnover of key cellular proteins. Often these regulatory proteases are either energy-dependent or intramembrane-cleaving. In archaea, two different types of energy-dependent proteases have been characterized: 20S proteasomes associated with proteasome-activating nucleotidases and membrane-associated Lon proteases. Interestingly, homologs of all three mechanistic classes of intramembrane cleaving proteases are widely distributed in archaea. Similar to their eucaryal and bacterial counterparts, members of these uncharacterized proteases might promote the controlled release of membrane-anchored regulatory proteins or liberate small peptide reporters and/or effectors that function in cell signaling. PMID- 16256424 TI - Molar absorptivities of 2,4-D, cymoxanil, fenpropidin, isoproturon and pyrimethanil in aqueous solution in the near-UV. AB - The absorption spectra of five pesticides, namely 2,4-dichloro-phenoxy acetic acid (2,4-D), cymoxanil, fenpropidin, isoproturon and pyrimethanil, have been measured in aqueous solution using a set-up consisting of two parallel absorption cells coupled to a CCD detector. The absolute values of their molar absorptivity coefficients epsilon were determined in the wavelength-range 240-344 nm with a deuterium-lamp at room temperature (298+/-2 K). Using the Beer-Lambert law, values of epsilon were also determined at 253.7 nm with a Hg-Lamp: epsilon = 145+/-14 for 2,4-D, epsilon = 7940+/-920 for cymoxanil, epsilon = 196+/-14 for fenpropidin, epsilon = 7330+/-880 for isoproturon, epsilon = 13200+/-1400 for pyrimethanil (in units of M(-1) cm(-1)). The quoted errors correspond to 2 sigma obtained from the least square fit analysis and the estimated systematic error of 5% due to the uncertainties in aqueous concentrations. For all the studied compounds, the absorbances measured were lower than 2.3 and did not exhibit any deviation from the Beer-Lambert's law. Our experimental data are discussed and compared to UV spectra of similar molecules when such data were available in the literature. Based on their UV spectra and the calculated fractions of these pesticides in the aqueous phase, their direct photolysis under sunlight environment could occur, except may be for fenpropidin, either in water surfaces or in aqueous droplets contained in the atmospheric clouds. PMID- 16256425 TI - Nonalcoholic steatohepatitis in Asia-Oceania. AB - Metabolic syndrome, i.e., obesity, hypertension, hyperlipidemia and insulin resistance with hyperinsulinemia is a new disease entity prevailing worldwide and nonalcoholic steatohepatitis is believed to be a hepatic expression of this syndrome. Nonalcoholic steatohepatitis is prevalent and well characterized in Caucasians, however, little is known about nonalcoholic steatohepatitis in Asia Oceania, because obesity has not been frequent in these countries. Obesity is sure to be a serious social problem in Asia-Oceania in the next two decades and we need to prevent the increase of nonalcoholic steatohepatitis. For that purpose, we need to know much about not only nonalcoholic steatohepatitis but also ourselves, as this syndrome is believed to be based not only on genetic but also environmental background. PMID- 16256426 TI - Sensitization and habituation of AMH and C-fiber related percepts of repetitive radiant heat stimulation. AB - OBJECTIVE: Pain perception involves neuronal plasticity at peripheral and central stages, resulting in sensitization or habituation, depending on intensity and temporal features of stimulation. Concurrent assessment of perceptual change over different time spans is therefore important for understanding the dynamics of pain processing. METHODS: A new psychophysical procedure was established to assess sensitization and habituation during repetitive radiant heat stimulation. Short-term perceptual change (<1 min) during trials with 10 stimuli applied at 3 frequencies (0.2-0.6-1.8 Hz) and 3 intensities was assessed for AMH-II or C-fiber related percepts. Perceptual changes were monitored for medium-term (1-15 min) and for long-term (15-90 min) time spans. RESULTS: Short-term sensitization occurred only at frequencies above 0.3 Hz and was affected by both stimulus frequency and intensity, but the AMH-fiber related sensitization depended on intensity only above 0.6 Hz. Multiple stimulation of the same skin area during medium-term time spans resulted in habituation. No long-term perceptual changes occurred. CONCLUSIONS: The procedure permits concurrent assessment of short-term sensitization and medium-term habituation, assumed to be related to spinal windup and cutaneous nociceptive fiber fatigue, respectively. SIGNIFICANCE: The method is suitable for quantitative sensory testing of dynamic pain processing over different time spans, relevant in clinical testing of pain and in drug assessment. PMID- 16256427 TI - Simultaneous multilobar electrocorticography (mEcoG) and scalp electroencephalography (scalp EEG) during intracranial vascular surgery: a new approach in neuromonitoring. AB - OBJECTIVE: Intraoperative neuromonitoring for intracranial vascular surgery is primarily aimed at detecting early ischemic changes to prevent subsequent infarction. Despite various neurophysiological approaches detection of early and focal ischemic changes remains difficult. This study explores the feasibility and sensitivity of intraoperative monitoring using surface EEG (scalp EEG) and multilobar Electrocorticography (mEcoG) recording during intracranial vascular procedures. METHODS: About 21 recordings were acquired in 20 patients undergoing craniotomies for intracranial aneurysms (17), superficial temporal-middle cerebral artery bypass (twice in the same patient) and arteriovenous malformation (2). The recording of scalp EEG (needle electrodes) and EcoG was performed (cupules electrodes) during all of the surgery. Signal was visually analyzed online and using spectral analysis software offline. RESULTS: Good recordings were obtained in all cases, without adding any procedural morbidity. The most common abnormalities on mEcoG were high frequency waves (23-37 Hz; HF-beta3), which appeared just after vascular occlusion and were occasionally followed by slow waves or burst suppression pattern. This focal pattern was seen in a majority of cases (20/21) on the mEcoG, but only in 4 out of 21 cases on the EEG. CONCLUSIONS: Multi-lobe (mEcoG) recording is feasible during craniotomies and detects earlier and more EEG pattern variation than surface EEG monitoring during intracranial vascular manipulations. The authors discuss the high sensitivity of this technique to ischemic changes. SIGNIFICANCE: By detecting earlier and more focal changes than scalp EEG, mEcoG may favor during surgery an increase in interactive strategies and reduction of deleterious event. PMID- 16256428 TI - Transcranial magnetic and electrical stimulation compared: does TES activate intracortical neuronal circuits? AB - OBJECTIVE: To determine whether, and under which conditions, transcranial electrical stimulation (TES) and transcranial magnetic stimulation (TMS) can activate similar neuronal structures of the human motor cortex, as indicated by electromyographic recordings. METHODS: Focal TMS was performed on three subjects inducing a postero-anterior directed current (p-a), TES with postero-anteriorly (p-a) and latero-medially (l-m) oriented electrodes. We analyzed the onset latencies and amplitudes (single-pulse) and intracortical inhibition and excitation (paired-pulse). RESULTS: TMS p-a and TES p-a produced muscle responses with the same onset latency, while TES l-m led to 1.4-1.9 ms shorter latencies. Paired-pulse TMS p-a and TES p-a induced inhibition at short inter-stimulus intervals (ISI) (maximum: 2-3 ms) and facilitation at longer ISIs (maximum: 10 ms). No inhibition but a strong facilitation was obtained from paired-pulse TES l m (ISIs 1-5 ms). CONCLUSIONS: Our findings support the hypothesis, that current direction is the most relevant factor in determining the mode of activation for both TMS and TES: TMS p-a and TES p-a are likely to activate the corticospinal neurons indirectly. In contrast, TES l-m may preferentially activate the corticospinal fibres directly, distant of the neuronal body. SIGNIFICANCE: TES is a suitable tool to induce intracortical inhibition and excitation. PMID- 16256430 TI - Auditory cortical tuning to statistical regularities in phonology. AB - OBJECTIVE: Ample behavioral evidence suggests that distributional properties of the language environment influence the processing of speech. Yet, how these characteristics are reflected in neural processes remains largely unknown. The present ERP study investigates neurophysiological correlates of phonotactic probability: the distributional frequency of phoneme combinations. METHODS: We employed an ERP measure indicative of experience-dependent auditory memory traces, the mismatch negativity (MMN). We presented pairs of non-words that differed by the degree of phonotactic probability in a modified passive oddball design that minimizes the contribution of acoustic processes. RESULTS: In Experiment 1 the non-word with high phonotactic probability (notsel) elicited a significantly enhanced MMN as compared to the non-word with low phonotactic probability (notkel). In Experiment 2 this finding was replicated with a non-word pair with a smaller acoustic difference (notsel-notfel). An MMN enhancement was not observed in a third acoustic control experiment with stimuli having comparable phonotactic probability (so-fo). CONCLUSIONS: Our data suggest that auditory cortical responses to phoneme clusters are modulated by statistical regularities of phoneme combinations. SIGNIFICANCE: This study indicates that the language environment is relevant in shaping the neural processing of speech. Furthermore, it provides a potentially useful design for investigating implicit phonological processing in children with anomalous language functions like dyslexia. PMID- 16256429 TI - Pain and calf hypertrophy associated with spontaneous repetitive discharges treated with botulinum toxin. AB - OBJECTIVE: To report a case of neurogenic calf hypertrophy associated with root lesion responsive to botulinum toxin (BTX). In addition, we systematically review the literature for cases of muscle hypertrophy associated with radiculopathy. METHODS: A 56-year-old man with a previous history of S1 root lesion presented with marked calf hypertrophy, muscle pain, cramps, fasciculations, and increased CK level. Magnetic resonance imaging and muscle biopsy confirmed true muscle hypertrophy. Electromyography (EMG) showed signs of chronic denervation reinnervation in the right S1 myotome, with greatly increased spontaneous activity (SA) (fasciculations and complex repetitive discharges). Botox(R) was injected in 3 occasions on 4 sites on the right gastrocnemius (20U per site). Symptoms, CK level and SA on EMG (quantified) were evaluated. RESULTS: The patient improved clinically, CK level and SA steadily decreased. CONCLUSIONS: The literature review failed to identify a unique causative factor for muscle hypertrophy related to neurogenic lesions. BTX treatment was effective in our patient with muscle hypertrophy associated with SA. SIGNIFICANCE: We provide further evidence that spontaneous activity secondary to nerve lesion may cause focal muscle hypertrophy. Treatment with BTX should be considered in these conditions. PMID- 16256431 TI - Silent EEG seizures and ictal seizures in the suppression period of neonatal burst-suppression records. PMID- 16256432 TI - Heart rate variability in comatose and brain-dead patients. PMID- 16256434 TI - Immersion in warm water induces improvement in cardiac function in patients with chronic heart failure. AB - BACKGROUND: The effects of immersion and training of patients with chronic heart failure (CHF) in warm water has not been thoroughly investigated. The aim of this study was to assess the acute hemodynamic response of immersion and peripheral muscle training in elderly patients with CHF. METHODS: Thirteen CHF patients and 13 healthy subjects underwent echocardiography on land and in a temperature controlled swimming pool (33-34 degrees C). RESULTS: Rest. Heart rate decreased (CHF, p=0.01; control, p=0.001) and stroke volume increased (CHF, p=0.01; control, p=0.001) during water immersion in both groups, with no change in systolic or diastolic blood pressure. Ejection fraction (p<0.05) and transmitral Doppler E/A ratio (p=0.01) increased in the CHF group, with no changes in left ventricular volumes. The healthy subjects had similar responses, but also displayed an increase in cardiac output (p<0.01) and left ventricular volumes (p<0.001). Exercise. Cardiac output and systolic blood pressure increased significantly in water, in both groups. CONCLUSION: A general increase in early diastolic filling was accompanied by a decrease in heart rate, leading to an increase in stroke volume and ejection fraction in most patients with CHF during warm water immersion. These beneficial hemodynamic effects might be the reason for the previously observed good tolerability of this exercise regime. PMID- 16256436 TI - Epidemic of tuberculosis in the former Soviet Union: social and biological reasons. AB - The tuberculosis epidemic was evidenced in the states of the former Soviet Union at the end of the 20th century. The increase of epidemiological indicators for tuberculosis was accelerated by socio-economic changes in the society. Laboratory analysis also revealed new biological properties of the Mycobacterium tuberculosis isolates in the region, in particular the high level of resistance to antibiotics. Tuberculosis in Russia and the other newly independent states has been caused by M. tuberculosis strains of the W-Beijing family in recent years. These bacteria have rapidly developed resistance to anti-tuberculosis drugs, as a result of prolonged exposure of the patients to inappropriate treatment. The epidemic of drug-resistant tuberculosis caused by the W-Beijing strains represents an important threat to the health of the populations in the newly independent states. It may also adversely affect the global trends in tuberculosis, as there is a clear risk that these drug-resistant strains can spread to other communities outside the former Soviet Union. PMID- 16256435 TI - Value of dual testing for identifying tuberculous infection. AB - SETTING: A rural community in Chingleput district in Tamil Nadu state in south India. OBJECTIVE: To determine the value of dual testing with PPD-S and PPD-B for identifying subjects with a tuberculous infection. DESIGN: About 240,000 subjects in rural south India, all of whom were tested initially with PPD-S and PPD-B, were followed up for 15 years, mainly by total population survey once in every 212 years. The incidence of culture-positive tuberculosis was estimated using life-table technique. RESULTS: Among 17,530 subjects with an intermediate reaction (8-11 mm) to PPD-S at intake, 285 with an induration to PPD-S exceeding the induration to PPD-B by at least 2mm, had a significantly higher incidence of culture-positive tuberculosis than the remaining (154 and 93 per 100,000), and similarly 481 who had an induration of <10mm to PPD-B compared to those with >or=10 mm (131 and 93 per 100,000). These subjects may be regarded as having a tuberculous infection. Infection with non-tuberculous mycobacteria conferred protection of about 30% against the development of tuberculosis over a 15-year period. CONCLUSION: In subjects with an intermediate reaction (8-11 mm) to PPD-S, dual testing with PPD-B enabled identification of those with a tuberculous infection. Most of the reactions were due to non-tuberculous mycobacteria. PMID- 16256437 TI - Oral solid lipid nanoparticle-based antitubercular chemotherapy. AB - The present study was planned to evaluate the chemotherapeutic potential of oral solid lipid nanoparticles (SLNs) incorporating rifampicin, isoniazid and pyrazinamide against experimental tuberculosis. The SLNs were prepared by the "emulsion solvent diffusion" technique with an encapsulation efficiency of 51+/ 5% for rifampicin, 45+/-4% for isoniazid and 41+/-4% for pyrazinamide. Following a single oral administration to mice, therapeutic drug concentrations were maintained in the plasma for 8 days and in the organs (lungs, liver and spleen) for 10 days whereas free drugs were cleared by 1-2 days. In M. tuberculosis H37Rv infected mice, no tubercle bacilli could be detected in the lungs/spleen after 5 oral doses of drug loaded SLNs administered at every 10th day whereas 46 daily doses of oral free drugs were required to obtain an equivalent therapeutic benefit. Thus, SLN based antitubercular drug therapy forms a sound basis for reducing dosing frequency and improving patient compliance for better management of tuberculosis. PMID- 16256438 TI - A new site-specific integration system for mycobacteria. AB - Site-specific integration into the mycobacterial chromosome can produce stable transformants useful for understanding pathogenesis. However, gene expression can be problematic at certain sites of integration. We have used the Streptomyces phiC31 integration system to integrate vector DNA into Mycobacterium smegmatis, M. bovis BCG, and M. tuberculosis through site-specific recombination. A single dominant insertion site was found in M. smegmatis, as previously reported. Three different insertion sites were found in M. bovis BCG. In M. smegmatis, integrated vectors appear to be far more stable than episomal plasmids during unselected passage in vitro, although excision products are detectable. Plasmids based on the phiC31 integration system could make useful tools for the study of mycobacterial genetics. PMID- 16256439 TI - Correlating sequential homology of Mce1A, Mce2A, Mce3A and Mce4A with their possible functions in mammalian cell entry of Mycobacterium tuberculosis performing homology modeling. AB - OBJECTIVE: The striking homology of the Mycobacterium tuberculosis mammalian cell entry operons (mce1, mce2, mce3 and mce4) with other mycobacterial species and the proposed role of the mammalian cell entry protein 1A (Mce1A) of M. tuberculosis to facilitate invasion of host cells have led us to look into the finer details of these proteins in order to better understand their structure function relationship. DESIGN: We performed sequential alignments and secondary structure predictions of Mce1A, Mce2A, Mce3A and Mce4A, and compared these results with results from homology modeling by fold prediction and threading. RESULTS: Sequential alignments showed that Mce1A and Mce2A are highly homologous, close to 70%, while the other combinations gave only about 30% similarities. The major parts of the proteins aligned without gaps and there were striking similarities by secondary structure predictions indicating that the proteins would have similar folds and to be alpha/beta proteins like the previously reported Mce1A model based on Colicin N. Fold prediction showed that the best templates for Mce2A were substrate-binding domain of DnaK and slow processing precursor penicillin acylase from Escherichia coli while the alpha-domains of Mce3A and Mce4A could both be modeled using the cytoplasmic domain of serine chemotaxis receptor as template. CONCLUSION: Although different templates had to be used to model the MceA proteins, functional information may be derived that is relevant for their overall function in M. tuberculosis. The beta-domain is probably involved in binding with the receptors on target cells while the alpha domain is more likely to be involved in pore formation. As predicted from the folds, Mce3A and Mce4A model structures indicate a lipid bound conformation and therefore may be required in signaling events of the mammalian cell entry process. PMID- 16256440 TI - Deciphering the genes involved in pathogenesis of Mycobacterium tuberculosis. AB - We had previously reported identification of the virS gene of Mycobacterium tuberculosis (Rv3082c) belonging to the AraC family of transcriptional regulators. The 7 genes (Rv3083-Rv3089) which are present divergently to virS (Rv3082c) constitute an operon designated as the mymA operon. Further investigation on the regulation of this operon showed that transcription of the mymA operon is dependent on the presence of VirS protein. A 4-fold induction of the mymA operon promoter occurs specifically in the wild type M. tuberculosis and not in the virS mutant of M. tuberculosis (M.tbDeltavirS) when exposed to acidic pH. Expression of the mymA operon was also induced in infected macrophages by ten fold over a six-day period. Bioinformatic analysis suggested the involvement of these proteins in the modification of fatty acids required for cell envelope. This was supported by altered colony morphology and cell envelope structure and increased accumulation of C24/C26 fatty acids by M.tbDeltavirS in comparison to the wild type strain. Thus, induction of mymA operon can play an important role in remodeling the envelope of intracellular M. tuberculosis under acidic conditions. Genomic analysis of M. tuberculosis revealed the presence of two tyrosine phosphatase genes--mptpA (Rv2234) and mptpB (Rv0153c). We have characterized both the tyrosine phosphatases of M. tuberculosis. To evaluate the role of MptpB in the pathogenesis of M. tuberculosis we have disrupted mptpB in the genome of M. tuberculosis. The wild type as well as mptpB mutant strain were comparable in their ability to infect and survive in the resting macrophages. However, the mptpB mutant strain was more sensitive to killing as compared to the wild type strain by IFN-gamma activated macrophages. In guinea pig model of tuberculosis an approximately 70-fold reduced bacillary load was observed in the spleen of the animals infected with mptpB mutant strain as compared to the bacillary load in animals infected with the wild type strain at 6 weeks post infection. These results suggest that mymA operon as well as mptpB gene of M. tuberculosis play an important role in the survival of the pathogen in the host. PMID- 16256441 TI - Protein kinase I of Mycobacterium tuberculosis: cellular localization and expression during infection of macrophage-like cells. AB - Protein kinase I of Mycobacterium tuberculosis, which has an unusual amino acid composition in its catalytic loop, displayed autophosphorylation and transphosphorylation activity. Immunoblot analysis of sub-cellular fractions of M. tuberculosis, using anti-PknI antibodies raised in rabbits, showed that PknI localizes to the bacterial cytosol. In contrast, PknA was membrane-bound. Relative expression of pknI, when measured by combining molecular beacons and RT PCR, decreased during infection of THP-1 human macrophages. Expression of pknA and pknB was upregulated during infection. Thus PknI represents a group of protein kinases that is distinct from the more extensively studied enzymes PknA and PknB. PMID- 16256442 TI - Comparing interventions in older unpartnered adults after myocardial infarction. AB - BACKGROUND: Older adults after myocardial infarction (MI) are a vulnerable group who may benefit from interventions to improve health outcomes. The use of a peer advisor or an advanced practice nurse (APN) to provide a self-efficacy intervention is a promising method of improving health outcomes after MI. AIMS: The purpose of this paper was to compare the effect of two self-efficacy interventions, a peer advisor and an APN, to a group who received standard care after MI. METHODS: The study was a three-group randomized clinical trial with a peer advisor intervention group, an APN intervention group, and a standard care group. Outcome data were collected in the hospital after MI and by telephone at 12 weeks after hospital discharge, after the interventions were completed. RESULTS: At 12 weeks after MI, there were no significant differences between the 3 groups in health outcomes. There were similar changes in self-efficacy for performing recovery behaviors, the actual performance of recovery behavior, physical and mental health across both intervention groups and the standard care group. CONCLUSIONS: Although the data did not validate the benefits of these self efficacy interventions, future efforts at identifying changes in health outcomes may need to use more discrete measurements that are more sensitive to changes in the older unpartnered adult after an MI. PMID- 16256443 TI - The effects of nitric oxide in acute lung injury. AB - Acute lung injury (ALI) is a common clinical problem associated with significant morbidity and mortality. Ongoing clinical and basic research and a greater understanding of the pathophysiology of ALI have not been translated into new anti-inflammatory therapeutic options for patients with ALI, or into a significant improvement in the outcome of ALI. In both animal models and humans with ALI, there is increased endogenous production of nitric oxide (NO) due to enhanced expression and activity of inducible NO synthase (iNOS). This increased presence of iNOS and NO in ALI contributes importantly to the pathophysiology of ALI. However, inhibition of total NO production or selective inhibition of iNOS has not been effective in the treatment of ALI. We have recently suggested that there may be differential effects of NO derived from different cell populations in ALI. This concept of cell-source-specific effects of NO in ALI has potential therapeutic relevance, as targeted iNOS inhibition specifically to key individual cells may be an effective therapeutic approach in patients with ALI. In this paper, we will explore the potential role for endogenous iNOS-derived NO in ALI. We will review the evidence for increased iNOS expression and NO production, the effects of non-selective NOS inhibition, the effects of selective inhibition or deficiency of iNOS, and this concept of cell-source-specific effects of iNOS in both animal models and human ALI. PMID- 16256444 TI - Electroencephalographic frequencies associated with heart changes in RR interval variability during paradoxical sleep. AB - Brain stem autonomic oscillators, hypothalamic and cortico-frontal centre, entrained by baroreceptor input, have been proposed as the control system of the heart rhythm. Recent reported results in animals suggested that the hippocampal theta waves might also participate as a heart rate modulator. A temporal correlation among the firing of neurons in the medulla, the R-wave of the electrocardiogram, hippocampal units, and theta rhythm was reported in guinea pigs. Our present aim is the analysis of the human electroencephalogram (EEG) frequencies power associated with changes in RR interval variability epochs during paradoxical sleep. We hypothesized that the differences in the human balance of the autonomic centres in sleep would be represented in the central nervous system by changes in the low-frequency EEG bands power. The heart rate analysis included 4 s windows, i.e., not considering the lowest component. The result was a consistent increment in the power of the paradoxical sleep delta and theta EEG bands during physiologic high heart RR interval variability epochs; no changes in the EEG bands power were found in the previous windows. The temporal correlation of heart RR interval variability and delta-theta EEG bands increases is proposed to represent a functional interaction when the control of specific centres fails or decreases during paradoxical sleep, a period mainly operating in an "open-loop" fashion. PMID- 16256446 TI - Numerical study of the electrical conductivity and polarization in a suspension of spherical cells. AB - The spatial distribution of electrical potential and current in a suspension of spherical cells under an applied electric field was numerically obtained using the equivalent circuit method (ECM). The effect of the proximity of the cells was studied in a set of simulations where the volumetric fraction varied from 0.24 to 0.66. The results show that the transmembrane potential for cells in the suspension is lower than the theoretically predicted value for a single dielectric membrane under a uniform electric field. It was also observed that as the volumetric fraction is increased, the transmembrane potential on the pole of the cells decreases linearly. Furthermore, the conductivity of the suspension was also observed to be a function of the volumetric fraction and this result is in a good agreement with the Maxwell's model for spherical particles suspended in a volume conductor. PMID- 16256445 TI - The role of corticotropin releasing factor and its antagonist, astressin, on micturition in the rat. AB - The purpose of this investigation was to evaluate the role of corticotropin releasing factor (CRF) on micturition. CRF is involved in the endocrine and central nervous system responses to stress and is also expressed in sites responsible for the control of micturition. In this investigation, cystometric experiments were performed in awake and unrestrained Wistar rats and on Spontaneous Hypertensive Rats, which are used as a rodent model of detrusor overactivity and anxiety. In vitro effects of CRF were evaluated using strips of detrusor muscle in an organ bath preparation. CRF (6.0 microg) administered via intrathecal and intraperitoneal routes, but not intracerebroventricularly, lowered the micturition threshold. CRF reduced the intercontraction interval by 28% and 26% after intrathecal or intraperitoneal administration, respectively, and reduced micturition volume by 34.7% and 30.2%, respectively. In Wistar-Kyoto rats, 6.0 microg intrathecal CRF significantly reduced intercontraction interval (423 +/- 79 vs. 669 +/- 59 s) and micturition volume (0.30 +/- 0.04 vs. 0.69 +/- 0.07 ml) compared to controls that received saline vehicle. These effects were blocked by pretreatment with 6.0 mug intrathecal astressin, a potent CRF antagonist, demonstrating that the effects are CRF receptor mediated. In Spontaneous Hypertensive Rats, 6.0 mug intrathecal CRF was found to have minimal stimulatory effects on the bladder, whereas astressin reduced baseline detrusor overactivity. CRF had no direct contractile effects on detrusor muscle strips. These results demonstrate that in the absence of detrusor overactivity, CRF stimulates micturition when administered via the intrathecal or intraperitoneal routes. Further studies are needed to explore the possibility whether CRF antagonists are effective for detrusor overactivity and the overactive bladder syndrome. PMID- 16256447 TI - Obstructive sleep apnoea is improved by a prolonged treatment of gastrooesophageal reflux with omeprazole. AB - BACKGROUND: Obstructive sleep apnoea has been demonstrated to induce gastrooesophageal reflux through highly negative intrathoracic pressure during the attacks. However, we believe that gastrooesophageal reflux on its part may favour or aggravate the apnoea attacks. AIMS: We investigated whether the treatment of gastrooesophageal reflux with omeprazole is able to decrease apnoea attacks. PATIENTS AND METHODS: Twenty patients (mean age 55.4 years, range 49-73 years; 17 males) with confirmed obstructive sleep apnoea at overnight polysomnography and pathological gastrooesophageal reflux at ambulatory 24-h oesophageal pHmetry were asked to note in a diary the occurrence of apnoea attacks for a basal period of 4 weeks. Subsequently, the patients were randomly and in a double-blind manner treated with omeprazole 20mg (10 patients, group A) or placebo (10 patients, group B) by giving 1 cp 30 min before breakfast and 1 cp 30 min before dinner for another 6 weeks with a diary documentation. The results were averaged weekly and over the entire treatment duration and a statistical comparison was made between the groups and within each group before and after treatment. RESULTS: The mean weekly frequency of apnoea attacks of group A during the entire period of treatment with omeprazole was significantly decreased with respect to the basal period and was significantly lower than that of group B. The weekly frequency of apnoea attacks in group A started to be significantly lower from the third week than the corresponding values of both group B and the basal period, reaching a decrease of about 73% in the sixth week. CONCLUSION: The occurrence of apnoea attacks progressively decreased during the treatment of gastrooesophageal reflux with omeprazole. This fact suggests that gastrooesophageal reflux may play a role in triggering and/or worsening obstructive sleep apnoea. PMID- 16256448 TI - Non-destructive studies of tissue-engineered phalanges by magnetic resonance microscopy and X-ray microtomography. AB - One of the intents of tissue engineering is to fabricate biological materials for the augmentation or replacement of impaired, damaged, or diseased human tissue. In this context, novel models of the human phalanges have been developed recently through suturing of polymer scaffolds supporting osteoblasts, chondrocytes, and tenocytes to mimic bone, cartilage, and tendon, respectively. Characterization of the model constructs has been accomplished previously through histological and biochemical means, both of which are necessarily destructive to the constructs. This report describes the application of two complementary, non-destructive, non invasive techniques, magnetic resonance microscopy (MRM) and X-ray microtomography (XMT or quantitative computed tomography), to evaluate the spatial and temporal growth and developmental status of tissue elements within tissue-engineered constructs obtained after 10 and 38 weeks of implantation in athymic (nude) mice. These two times represent respective points at which model middle phalanges are comprised principally of organic components while being largely unmineralized and later become increasingly more mineralized. The spatial distribution of mineralized deposits within intact constructs was readily detected by XMT (qCT) and was comparable to low intensity zones observed on MRM hydration maps. Moreover, the MRM-derived hydration values for mineralized zones were inversely correlated with mineral densities measured by XMT. In addition, the MRM method successfully mapped fat deposits, collagenous tissues, and the hydration state of the soft tissue elements comprising the specimens. These results support the application of non-destructive, non-invasive, quantitative MRM and XMT for the evaluation of constituent tissue elements within complex constructs of engineered implants. PMID- 16256449 TI - The effects of vanadium treatment on bone in diabetic and non-diabetic rats. AB - Vanadium-based drugs lower glucose by enhancing the effects of insulin. Oral vanadium drugs are being tested for the treatment of diabetes. Vanadium accumulates in bone, though it is not known if incorporated vanadium affects bone quality. Nine- to 12-month-old control and streptozotocin-induced diabetic female Wistar rats were given bis(ethylmaltolato)oxovanadium(IV) (BEOV), a vanadium based anti-diabetic drug, in drinking water for 12 weeks. Non-diabetic rats received 0, 0.25 or 0.75 mg/ml BEOV. Groups of diabetic rats were either untreated or treated with 0.25-0.75 mg/ml BEOV as necessary to lower blood glucose in each rat. In diabetic rats, this resulted in a Controlled Glucose group, simulating relatively well-managed diabetes, and an Uncontrolled Glucose group, simulating poorly managed diabetes. Plasma insulin, glucose and triglyceride assays assessed the diabetic state. Bone mineral density (BMD), mechanical testing, mineral assessment and histomorphometry measured the effects of diabetes on bone and the effects of BEOV on non-diabetic and diabetic bone. Diabetes decreased plasma insulin and increased plasma glucose and triglycerides. In bone, diabetes decreased BMD, strength, mineralization, bone crystal length, and bone volume and connectivity. Treatment was effective in incorporating vanadium into bone. In all treated groups, BEOV increased osteoid volume. In non diabetic bone, BEOV increased cortical bone toughness, mineralization and bone formation. In controlled glucose rats, BEOV lowered plasma glucose and improved BMD, mechanical strength, mineralization, bone crystal length and bone formation rate. In poorly controlled rats, BEOV treatment slightly lowered plasma glucose but did not improve bone properties. These results suggest that BEOV improves diabetes-related bone dysfunction primarily by improving the diabetic state. BEOV also appeared to increase bone formation. Our study found no negative effects of vanadium accumulation in bone in either diabetic or non-diabetic rats at the dose given. PMID- 16256450 TI - Systemic bone loss and induction of coronary vessel disease in a rat model of chronic inflammation. AB - Clinically, osteopenia or low bone mass has been observed in a variety of chronic inflammatory diseases, and elevated proinflammatory mediators have implicated this process. The purpose of this study was to develop an in vivo model of bone loss induced by chronic systemic inflammation. Time-release pellets designed to deliver one of three doses of LPS: Low (3.3 microg/day), High (33.3 microg/day), or Placebo over 90 days, were implanted subcutaneously in 3-month-old male Sprague-Dawley rats (n = 8/group). Neutrophil counts, indicative of ongoing inflammation, were elevated (P < 0.05) in both LPS groups at 30 days post-implant and remained significantly elevated in the High dose throughout the 90-day study period. At the end of the study, bone loss occurred in the femur as indicated by decreased bone mineral density (BMD) in both LPS-treated groups, but vertebral BMD was reduced in the High dose animals only. Microcomputed tomography revealed that trabecular bone volume (BV/TV) of the proximal tibial metaphysis tended to be reduced in the High dose LPS group. Deleterious effects on trabecular number (TbN) and trabecular separation (TbSp) were observed in both LPS-treated groups, but only the High dose group reached statistical significance. These alterations in trabecular microarchitecture resulted in compromised biomechanical properties. No changes in cortical thickness, porosity, or area of the tibia midshaft were evident at either dose of LPS. Up-regulation of the proinflammatory mediators, cyclooxygenase (COX)-2, interleukin (IL)-1, and tumor necrosis factor (TNF)-alpha was demonstrated in the metaphyseal region where the deleterious effects of LPS were observed. In addition to these alterations in bone, trichrome staining indicated changes in the coronary arterioles, consistent with vascular disease. Utilization of a LPS time-release pellet appears to provide an in vivo model of chronic inflammation-induced bone loss and a potentially novel system to study concurrent development of osteopenia and vascular disease. PMID- 16256453 TI - Modeling the adsorption of Cd(II) onto Muloorina illite and related clay minerals. AB - The adsorption of Cd(II) onto goethite, kaolinite, and illite was measured as a function of pH (adsorption edges) and concentration (adsorption isotherms) at 25 degrees C. As the pH was increased, adsorption onto goethite occurred mainly in the pH range 5.5-8, whereas adsorption onto kaolinite occurred in two stages, separated by a plateau in the pH region 5.5 to 7. Adsorption onto illite increased steadily as the pH was increased, with far less Cd(II) adsorbing onto illite than onto goethite or kaolinite per m(2) of mineral surface area. Potentiometric titrations of suspensions of each mineral, with and without Cd(II) present, were also completed. Results from all three types of experiments were modeled using an extended constant- capacitance surface complexation model. The reactions [Formula: see text] [Formula: see text] and [Formula: see text] best described Cd(II) adsorption onto goethite, while [Formula: see text] and [Formula: see text] best described Cd(II) adsorption onto kaolinite. A combination of the first, second, and fourth of these reactions best fitted the data for Cd(II) adsorption onto illite. In each case the model fitted all experimental data well. The results suggest that adsorption onto the variable charge (SOH) sites on illite more closely resembles adsorption onto goethite than onto kaolinite. PMID- 16256454 TI - Protective effect of small amounts of sodium dodecyl sulfate on the helical structure of bovine serum albumin in thermal denaturation. AB - In the presence of sodium dodecyl sulfate (SDS), the secondary structure of bovine serum albumin (BSA) was almost protected against thermal denaturation above 50 degrees C, where the structural change became irreversible. Beyond 30 degrees C, the helicity (66%) of the protein sharply decreased with rise of temperature. In response to this, the proportions of beta-structure and random coil increased. The helicity and the beta-structural proportion were 44% and 13% at 65 degrees C, respectively. The protective effect was observed upon the coexistence of SDS of extremely low concentrations: the molar ratio of [SDS]/[BSA] of 15 was enough to induce the maximal protective effect on the helical structure of the protein. The maximal protected helicity was 58% at 65 degrees C, increasing to 64% upon cooling down to 25 degrees C. This protective effect became greater with an increase of chain length of alkyl sulfate ion. On the other hand, a cationic surfactant did not protect the BSA structure at all against the thermal denaturation. This protective effect was characterized by the specific amphiphilic nature of anionic surfactant. Such an anionic surfactant is considered to protect the protein structure by building bridges between particular nonpolar residues and particular positively charged residues located on different loops of the protein. PMID- 16256455 TI - Well-defined fluorescent particles of ZnS:Cu. 1. Preparation and effects of annealing. AB - As an application of the gel-sol procedure for the synthesis of monodispersed particles in large quantities, uniform particles of ZnS:Cu were prepared by different methods from concentrated Zn-chelate solutions of nitrilotriatic acid (NTA) with thioacetamide (TAA), in which the dopant copper ion was introduced as a coexisting mixed chelate with Zn-NTA (Method A), a Cu-NTA complex in a separate solution continuously added to a system of Zn-NTA and TAA (Method B), or a Cu(NO3)2 solution infiltrated into a dry powder of ZnS prepared by Method A (Method C). All powders thus prepared were annealed before the test of fluorescent properties. The main roles of annealing were to remove the grain boundaries of such particles as nonradiative recombination centers of excited electrons and holes and to achieve uniform distribution of dopants. However, the initial intraparticle distribution of the dopant, controlled by the different methods, decisively affected the fluorescent particles, even after the conversion of the polycrystalline particles to single-crystal particles by annealing. PMID- 16256456 TI - Synthesis and aqueous solution properties of polyelectrolyte-grafted silica particles prepared by surface-initiated atom transfer radical polymerization. AB - A range of polyelectrolyte-grafted silica particles have been prepared by grafting suitable initiators onto near-monodisperse, 304-nm-diameter silica particles using siloxane chemistry, followed by surface-initiated atom transfer radical polymerization (ATRP) of four ionic vinyl monomers, namely sodium 4 styrenesulfonate (SStNa), sodium 4-vinylbenzoate (NaVBA), 2-(dimethylamino)ethyl methacrylate (DAM), and 2-(diethylamino)ethyl methacrylate (DEA) in protic media. The resulting polyelectrolyte-grafted silica particles were characterized using dynamic light scattering (DLS), thermogravimetric analysis (TGA), X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM), helium pycnometry, and diffuse reflectance infrared Fourier transfer spectroscopy (DRIFTS). The TGA results indicated that the polyelectrolyte contents of the silica particles could be varied from 0.6% to 6.0% in weight. SEM studies revealed several surface morphologies for the grafted polyelectrolytes and XPS analysis of the particle surface also provided good evidence for surface grafting. Combined aqueous electrophoresis and DLS studies confirmed that these polyelectrolyte-grafted silica particles had pH-dependent colloid stabilities, as expected. Cationic polyelectrolyte-grafted silica particles were colloidally stable at low or neutral pH, but became aggregated at high pH. Conversely, anionic polyelectrolyte-coated silica particles became unstable at low pH. It was found that the rate of surface-initiated ATRP was substantially slower than the analogous solution polymerization. Finally, there was some evidence to suggest that, at least in some cases, a significant fraction of polymer chains became detached from the silica particles during polymerization. PMID- 16256457 TI - A broad frequency range dielectric spectrometer for colloidal suspensions: cell design, calibration, and validation. AB - Electrode polarization complicates low-frequency measurements of the dielectric response of electrolyte solutions and colloidal suspensions. To deal with this longstanding problem, a new dielectric cell was developed along with a model based on the standard electrokinetic theory. The parallel plate cell utilizes a thin chamber that is easily filled and emptied; different chamber thicknesses are readily accommodated. The analytical form of the theoretical impedance model makes data analysis straightforward. Using standard electrolytes, the device and the theoretical model were tested over a wide range of frequencies for several electrolyte concentrations. Excellent agreement was found between the theory and the experimental data. The methodology developed to account for polarization effects exhibits a significant improvement over the conventional approaches and points up a deficiency in often-used equivalent circuit models. PMID- 16256458 TI - Structural-chemical disorder of manganese dioxides 1. Influence on surface properties at the solid-electrolyte interface. AB - Relationships between lattice parameters of manganese dioxides and their surface properties at the solid-aqueous solution interface were investigated. The studied series ranged from ramsdellite to pyrolusite and encompassed disordered MD samples. The structural model used takes into account structural defects: Pr (rate of pyrolusite intergrowth) and Tw (rate of microtwinning). Water adsorption isotherms showed that the cross sectional area of water molecules adsorbed in the first monolayer is positively correlated to Pr. Titration of the surface charge of the MD series evidenced a positive linear relationship between the PZC and Pr (Pr=0, Tw=0, PZC=1 for ramsdellite; Pr=1, Tw=0, PZC=7.3 for pyrolusite; gamma-MD with intermediate values of Pr (0.2 to 0.45) have increasing PZC values). The rate of microtwinning appeared as a secondary factor for the increase of the PZC. The above correlations are explained by the chemical defects at the origin of the structural disorder, respectively Mn(3+)/Mn4+ substitution for Pr and Mn vacancies for Tw, which result in proton affinity and thus in increased PZC. The experimental results are compared with data collected in the literature for manganese dioxides as well as for dioxides of transition elements with tetragonal structure. PMID- 16256459 TI - Experimental and theoretical study of the displacement process between two electrolyte solutions in a microchannel. AB - Displacement of one electrolyte solution by another in a microchannel is required in many biolab chip devices. The objective of this paper is to develop a better understanding of the displacement process between two electrolyte solutions under an applied electric field in a cylindrical microchannel in terms of the traveling distance of the interface between these two electrolyte solutions. In order to develop a general model to predict the location of the interface, two different situations are considered; one model assumes the presence of a sharp interface between the two solutions and the other model considers a mixing zone between the two solutions. Carefully conducted experiments were carried out to obtain the current-time relationship, which is used in the model to predict the location of the interface. In these experiments, deionized ultrafiltered water (DIUF water), 10 mM KCl, 0.1 mM KCl, and 0.1 mM LaCl3 solutions were used as the testing liquids. Polyamide-coated silica capillary tubes of internal diameter 100 mum and length 10 cm were employed in this study. The relationship between traveled distance of the interface and time was predicted by a developed model based on the measured current-time relationship for such a displacement process under a constant applied electric field. The characteristics of the nonlinear change of the traveling distance with the time were also discussed in this paper. PMID- 16256460 TI - Film drainage between two surfactant-coated drops colliding at constant approach velocity. AB - The drainage of the intervening continuous phase film between two drops approaching each other at constant velocity under the influence of insoluble surfactant is investigated. The mathematical model to be solved is a coupled pair of fourth-order nonlinear partial differential equations which arise from the relationships governing the evolution of the film thickness and the surfactant interfacial concentration in the lubrication approximation. We adopt a simplified approach which uses lubrication theory to describe the flow within the drop, marking a departure from the conventional framework in which Stokes flow is assumed. When the model is solved numerically together with the relevant initial and boundary conditions, the results obtained are compared with those found in the literature using the "boundary integral" method to solve for the flow in the drop phase. The close agreement between the results inspires confidence in the predictions of the simplified approach adopted. The analysis on the effect of insoluble surfactant indicates that its presence retards the drainage of the film: The fully immobile interface limit is recovered even in the presence of a small amount of surfactant above a critical concentration; film rupture is either prolonged or prevented. The retardation of the film was attributed to gradients of interfacial tension which gave rise to the Marangoni effect. A study of the influence of various system parameters on the drainage dynamics was conducted and three regimes of drainage and possible rupture were identified depending on the relative magnitudes of the drop approach velocity and the van der Waals interaction force: Nose rupture, rim rupture, and film immobilization and flattening. Finally, the possibility of forming secondary droplets by encapsulating the continuous phase film into the coalesced drop at rupture was examined and quantified in light of these regimes. PMID- 16256462 TI - A quantitative approach to studying structures and orientation at self-assembled monolayer/fluid interfaces. AB - Self-assembled monolayers (SAMs) have become a standard tool for exploring surface interactions. Although well characterized, SAMs are known to undergo structural and conformational changes in the presence of solution, yet the ability to quantify these changes remains an obstacle due to limited analytical techniques. In this study, we determine changes in structure and conformation of CH3, OH, and COOH terminated hexadecanethiols on gold in water by means of a new technique known as evanescence reflection spectroscopy. This FTIR application, in conjunction with a semiempirical formalism, is capable of providing both qualitative and quantitative understanding of the molecular structure and orientation at the solid/liquid interface. PMID- 16256461 TI - Induced removal of dipalmitoyl phosphatidylcholine by the exclusion of fibrinogen from compressed monolayers at air/liquid interfaces. AB - The induced removal of dipalmitoyl phosphatidylcholine (DPPC) by the exclusion of fibrinogen from mixed DPPC/fibrinogen monolayers at compressed air/liquid interfaces was analyzed. The surface pressure-area hysteresis curves of the monolayers at interfaces were obtained by a Langmuir trough. The hysteresis curves of equilibrium fibrinogen adsorption layers suggest that fibrinogen desorption during the area compression stage became significant at a higher bulk concentration of 1000 ppm. For mixed monolayers of DPPC with fibrinogen, the fibrinogen molecules were expelled from the interface upon compression due to the presence of insoluble DPPC molecules. The squeeze-out of fibrinogen molecules evidently removed a significant number of DPPC molecules from the interface, with the extent depending on fibrinogen surface concentration. During the subsequent area expansion stage, fibrinogen molecules entered the interface and participated in the rise of surface pressure. The induced loss of free DPPC molecules at the interface by the expelled fibrinogen molecules during the area compression stage was then evaluated from the hysteresis curves. PMID- 16256463 TI - Irreversible temperature-induced changes in purple membranes studied by electrooptics. AB - The thermal stability of purple membranes is studied by electric light scattering. Information on the polarization mechanisms is obtained. Incomplete recovery of the initial electric state (i.e., of permanent dipole moment, p( perpendicular), and electric polarizability, gamma(fast, ||)) after the membranes are heated at temperatures above 60 degrees C is revealed. Additional slow polarizability components, gamma(slow, perpendicular) and gamma(slow, ||), relaxing at different characteristic frequencies than the fast longitudinal polarizability gamma(fast, ||) appear in the temperature range where the order disorder transition takes place. The slow polarizability gamma(slow, perpendicular) is probably connected with counterion displacement in the electrical double layer perpendicular to the disk plane. The results are important for understanding the polarization mechanisms and the origin of slow orienting moments. PMID- 16256464 TI - Studies of the effect of variation of blend ratio on permselectivity and heterogeneity of ion-exchange membranes. AB - Heterogeneous ion-exchange membranes (both cationic and anionic types) have been synthesized by solution casting techniques using polyvinyl chloride (PVC) as binder and ion-exchange resin (-300+400 mesh). The binder:resin ratio varied from 60:40 to 30:70. The transport behavior of the membranes has been evaluated chronopotentiometrically in sodium chloride (NaCl) solutions of different concentrations. The different parameters E(0) (potential drop across the membrane at the instant of application of current I), E(max) (maximum potential drop across the membrane after the application of current I), DeltaE (magnitude of the potential jump across the membrane at transition time tau), Itau(1/2), tau, etc., have been evaluated. The isoconductance points were determined and based on the microheterogeneous model proposed by Zabolotsky and Nikonenko (J. Membrane Sci. 79 (1993) 181) the distribution factors beta has been evaluated for both types of ion exchange membranes. The electroconductivity of the joint gel (kappa ) and pure gel phases (kappa ' ) has been determined. At any particular solution concentration the transport number as well as the permselectivity of membranes increases with increased resin content of the membrane. The microheterogeneity factor beta exhibits synchronization among the each set of four different membranes for both the cationic and anionic type. PMID- 16256465 TI - Removal of phosphate by aluminum oxide hydroxide. AB - The development and manufacture of an adsorbent to remove phosphate ion for the prevention of eutrophication in lakes are very important. The characteristics of phosphate adsorption onto aluminum oxide hydroxide were investigated to estimate the adsorption isotherms, the rate of adsorption, and the selectivity of adsorption. Phosphate was easily adsorbed onto aluminum oxide hydroxide, because of the hydroxyl groups. The adsorption of phosphate onto aluminum oxide hydroxide was influenced by pH in solution: the amount adsorbed was greatest at pH 4, ranging with pH from 2 to 9. The optimum pH for phosphate removal by aluminum oxide hydroxide is 4. The selectivity of phosphate adsorption onto aluminum oxide hydroxide was evaluated based on the amount of phosphate ion adsorbed onto aluminum oxide hydroxide from several anion complex solutions. It is phosphate that aluminum oxide hydroxide can selectively adsorb. The selectivity of phosphate onto aluminum oxide hydroxide was about 7000 times that of chloride. This result indicated that the hydroxyl groups on aluminum oxide hydroxide have selective adsorptivity for phosphate and could be used for the removal of phosphate from seawater. PMID- 16256466 TI - An alternative description of the interfacial energy of a liquid in contact with a solid. AB - Aqueous solutions and liquid metals in contact with other phases or components experience strong Coulomb forces at their interfaces. The interfacial energy of such liquids facing solid walls is given by the sum of the free surface energy of the liquid and the electrostatic energy of the diffusive electric double layer due to the wall surface charge density. As a consequence, the interfacial stress of the liquid at the wall appears to be an anisotropic stress tensor, which has a stress component of opposite sign normal to the surface, compared with its two components parallel to the surface. This alternative description of the interfacial energy makes it possible to predict static contact angles, once the wall surface charge density is known. This paper investigates the relationship between the long-range Coulomb effects in the ionic solution and the interfacial energy, and concentrates on experiments in which other surface effects are minimized and in which chemical reactions at the surface are absent or frozen. Data for water and mercury, as an example, fit well if the same surface charge density of the wall is assumed in both cases. As an application to micro fluid dynamics, the electroosmotic mobility, the streaming current, or the pressure drop of a flow through a microchannel can be predicted only from contact angle measurements done. PMID- 16256467 TI - Small angle neutron scattering study of sodium dodecyl sulfate micellar growth driven by addition of a hydrotropic salt. AB - The structures of aggregates formed in aqueous solutions of an anionic surfactant, sodium dodecyl sulfate (SDS), with the addition of a cationic hydrotropic salt, p-toluidine hydrochloride (PTHC), have been investigated by small angle neutron scattering (SANS). The SANS spectra exhibit a pronounced peak at low salt concentration, indicating the presence of repulsive intermicellar interactions. Model-independent real space information about the structure is obtained from a generalized indirect Fourier transformation (GIFT) technique in combination with a suitable model for the interparticle structure factor. The interparticle interaction is captured using the rescaled mean spherical approximation (RMSA) closure relation and a Yukawa form of the interaction potential. Further quantification of the geometrical parameters of the micelles was achieved by a complete fit of the SANS data using a prolate ellipsoidal form factor and the RMSA structure factor. The present study shows that PTHC induces a decrease in the fractional charge of the micelles due to adsorption at the micellar surface and consequent growth of the SDS micelles from nearly globular to rodlike as the concentration of PTHC increases. PMID- 16256468 TI - Different dispersion regions during the phase inversion of an ionomeric polymer water system. AB - Catastrophic phase inversion is induced by changing the phase ratio in a liquid liquid dispersion and is widely used during the dispersion stage in the production of aqueous polyurethane ionomer (PUI) colloids. In the work reported here, water was added to polyurethane ionomer prepolymer (PUIp) until the water became the continuous phase. Three different dispersion regions have been discovered by changing the ionic group content. Stable emulsions containing small polymer drops were produced in Region I. Stable coarse emulsions containing a mixture of drop structures were produced in Region II, but only temporary dispersions could be produced in Region III. Conductivity measurements could not always be used to detect the phase inversion points effectively because the PUIp was swollen by water. Therefore, torque change measurements have been used in conjunction with the conductivity measurements to detect the phase inversion points for all three dispersion regions. Scanning electron microscopy (SEM) and optical microscopy were used to obtain images of these dispersions in the different regions. A catastrophic phase inversion map is used to represent the changes that occur in the PUIp-W dispersions. This map is plotted using the ionic group content as the ordinate and water content (at the phase inversion points) as the abscissa. PMID- 16256469 TI - Oxidation of activated carbon: application to vinegar decolorization. AB - This article reports studies on the feasibility of increasing the decoloring capacity of a granular activated carbon (GAC) by using oxidation with air at 350 degrees C to modify its surface activity and porosity. The GAC, obtained from olive stones, had a maximum decolorization capacity of 92% for doses of 20 g/l, while the maximum decolorization capacity of the modified granular activated carbon (MGAC) was about 96% at a dose of 10 g/l. The increase in decoloring capacity is thought to be due to an increase in mesopore area (from 129 to 340 m2/g) in the MGAC. The maximum decoloring values and the doses needed to attain them are very close to values obtained in previous studies using coconut shell powder-activated carbon (94 and 98% for red and white vinegar for a dose of 10 g/l, respectively). PMID- 16256470 TI - An electron paramagnetic resonance study of Cu(II) sorbed on quartz. AB - The nature of the interaction among Cu(II), adsorbed water, and quartz surface was studied using electron paramagnetic resonance (EPR) spectroscopy. The EPR lineshape gave information concerning the motional status of sorbed Cu(II) that revealed its binding strength at the surface. Two distinct absorption lines of sorbed Cu(II), namely, the liquid-type and the solid-type signal, were simultaneously observed at the fully hydrated surface at room temperature. The absorption lines and the variation of their intensity with experimental and measurement conditions such as degree of hydration, pH, ionic strength, and surface coverage indicated that there exist three kinds of Cu(II) entities, the inner-sphere surface complex, the outer-sphere surface complex, and the surface precipitate on the quartz surface, and that their concentrations change with experimental conditions. The reversible conversion of the liquid-type signal to the solid-type one during the drying-wetting or freezing-melting of the surface suggested the development of multiple layers of adsorbed water molecules on the quartz surface. It is assumed that the innermost layer of the water layers contains the inner-sphere Cu(II) surface complexes, while the outer layers contain the outer-sphere complexes whose binding strength decreases outward with increasing distance from the surface. The result of this work suggests that the sorption mechanism of a metal cation on a given mineral surface; hence its mobility in the environment may change significantly with the solution pH, the ionic strength, and the surface coverage. PMID- 16256471 TI - Adsorption of cations onto the surfaces of silver nanoparticles. AB - The effects of cations on the absorption spectra of silver sols have been investigated by the UV-vis spectrometry and TEM. Experiments showed that injection of certain amounts of transition metal cations into silver sols resulted not only in the aggregation of silver nanoparticles but also in the appearance of a new band centered near 510 nm in the absorption spectra of silver sols. However, the new band was not observed in the presence of alkaline earth metal cations or the Mv2+ cations. The peak position of the new band depends on the nature as well as the concentration of metal cations used. Comparing the peak positions of the new bands, it was found that the new band induced by the injection of Cr3+ was red-shifted with respect to those induced by Cu2+, Zn2+, or the Cd2+ cations. It is reasonable that this band near 510 nm should be attributed to the coeffects of the adsorption of metal cations onto the surfaces of silver nanoparticles and the aggregation of silver nanoparticles. PMID- 16256472 TI - The changes in particle charge distribution during rapid growth of particles in the plasma reactor. AB - The changes in particle charging were investigated during the rapid growth of particles in the plasma reactor by the discrete-sectional model and the Gaussian charge distribution function. The particle size distribution becomes bimodal in the plasma reactor and most of the large particles are charged negatively, but some fractions of small particles are in a neutral state or even charged positively. As the particles accumulate in the plasma reactor, the amount of electrons absorbed onto the particles increases, while the electron concentration in the plasma decreases. As the mass generation rate of small particles (monomers) decreases or as the initial electron concentration increases, the electron concentration in the plasmas increases and the particle charge distribution is shifted in the negative direction and the fraction of particles charged negatively and the average number of electrons per particle increase. With the decrease in monomer diameter, the electron concentration decreases in the beginning of plasma discharge, but, later, increases. For high mass generation rate of monomers or for low initial electron concentration or for small monomer diameter, the fraction of particles in a neutral state increases and the particle size distribution becomes broader. PMID- 16256473 TI - Molecular configurations and orientations of hydrazine between structural layers of kaolinite. AB - Hydrazine is one of the most commonly used entraining agents to penetrate kaolinite, yet the mechanism of intercalation of kaolinite by hydrazine is still in debate. The objectives of this study are to investigate the possible molecular configurations and orientations of hydrazine in the interlayer of kaolinite and the configuration changes induced by water molecules. Water molecules increased the intercalation rate and caused the expansion of the intercalation complex from 0.96 to 1.03 nm. The kinetic effect was likely the result of breaking the self associations of hydrazine molecules and releasing more "free" hydrazine molecules for the intercalation. H-bonding caused large red shifts of the inner surface OH stretching bands from 3695 to 3626 cm(-1) in the 0.96-nm kaolinite hydrazine intercalation (KHI) complex and to 3570 and 3463 cm(-1) in the 1.03-nm KHI complex. The NH stretching bands of the hydrazine molecules in the KHI complexes became sharper and blue-shifted more than 20 cm(-1) compared with the free liquids. The symmetric NH vibrations at 3365 and 3310 cm(-1), and the NN vibration at 1092 cm(-1) became infrared inactive in the 0.96-nm KHI complex. The frequency of the SiO bands of the kaolinite in the 1.03-nm KHI complex was slightly lower than in the 0.96-nm KHI complex (5 cm(-1) shift). These IR band changes implied that hydrazine molecules have different configurations in the complexes: hydrazine molecules had an eclipsed form in the interlayer of the 0.96 nm KHI complex. The eclipsed configuration has a dipole moment of 3.31 D, which is higher than the gauche form (1.83-1.90 D). The molecule was oriented with the NN bond parallel or nearly parallel to the (001) surface of the mineral and the four H atoms of each hydrazine molecule reacted with the basal siloxane surface. When a suitable amount of water was present, it promoted the configuration change of the hydrazine molecules from the eclipsed form to the common gauche form. This gauche form was stabilized by transforming to a more polarized NH3NH tautomer structure (5.4 D). To promote an optimal interaction between hydrazine and the mineral surface, the NN bond of the hydrazine was tilted about 30 degrees from the (001) plane and caused the intercalation complex to expand from 0.96 to 1.03 nm. The eclipsed form and the tautomer were stabilized by the asymmetric interlayer environment of kaolinite. The two proposed models and reaction mechanisms match the high dipole moment requirement as found for other entraining agents. Further investigation is needed to confirm the exact configuration of hydrazine molecules and whether or not the tautomer exists. PMID- 16256474 TI - Electrorheological fluids based on glycerol-activated titania gel particles and silicone oil with high yield strength. AB - This article describes an electrorheological (ER) fluid based on glycerol activated titania organic-inorganic hybrid gel particles and silicone oil with high yield strength. Based on a physical picture of a water-activated ER system, glycerol that has a high dielectric constant and boiling point is in situ prepared in the amorphous titania gel during the sol-gel processing. A small amount of ionic surfactant hexadecyltrimethylammonium bromide (CTAB) is employed to enhance charge carriers in particles. FTIR and XRD techniques are used to determine the nature and structure of the hybrid gel. Rheology test results show that a large static yield stress greater than 12.6 kPa is obtained when 3 kV/mm dc electric field is applied. This value is close to the value predicted by H. Conrad (MRS Bull. 8 (1998) 35) in theory. Furthermore, dynamic shear stress as a function of shear rate and temperature is also investigated. This ER fluid exhibits strong temperature dependence and a wide working temperature range from 0 to 120 degrees C, while its leaking current density is still low. More interesting is that the glycerol content is demonstrated to have an influence on ER effect and temperature dependence. Measurement of the dielectric properties of ER fluids shows enhancement of the dielectric constant and dielectric loss due to addition of glycerol and a regular dependence of them on temperature, which well explains the strong ER effect. PMID- 16256476 TI - Studies on intrinsic ionization constants of Fe-Al-Mg hydrotalcite-like compounds. AB - The theoretical analysis of the intrinsic ionization constant (K(a2)(int)) of Fe Al-Mg hydrotalcite-like compounds (HTlc) possessing permanent charges was first performed using the double extrapolation method proposed by James et al. The theoretical permanent charge density (sigma(p,T)) of the HTlc sample was calculated from the crystal structure of HTlc, and the influence of sigma(p,T) on the K(a2)(int) was also examined. From the experimental results, these conclusions can be obtained: the zero point of the charge (pH(ZPC)) of Fe-Al-Mg HTlc increases with decreased Fe3+ content and increases with increased Mg2+ in the HTlc. The pK(a2)(int) of Fe-Al-Mg HTlc also increases with decreased Fe3+ and increased Mg2+ content in the sample; furthermore, the pK(a2)(int) of Fe-Al-Mg HTlc increases with decreased sigma(p,T). PMID- 16256475 TI - Structural, morphological, and magnetic study of nanocrystalline cobalt-nickel copper particles. AB - Nanocrystalline Co(x)Ni(y)Cu(100-x-y) particles were synthesized by the reduction of metal acetates in a mixture of polyol and Tween 80. Inductively coupled plasma (ICP) analysis revealed that the actual wt% of Co, Ni, and Cu in these nanoparticles was nearly the same as in the starting solutions. The structures of the particles were characterized by X-ray diffraction (XRD), transmission electron microscopy (TEM), selected area electron diffraction (SAED) spectroscopy, and vibrating sample magnetometry (VSM). The results of XRD and VSM confirmed that there was no metastable alloying in the particles. The particles were composites, consisting of nanoscale crystallites of face-centered cubic (fcc) Cu, face-centered cubic (fcc) Ni, and face-centered cubic (fcc) Co. During preparation the nucleation of Cu occurred first; then small Cu nuclei acted as cores for the precipitation of Co and Ni. The particles showed an increase in saturation magnetization (M(s)) as the concentration of Co or Ni in the particles was increased. The changes of both M(s) and coercivity of the particles with increasing annealing temperatures were studied. The coercivity of the particles was very high; it could reach as high as 489 Oe for Co34.3Ni31.2Cu34.5) . PMID- 16256477 TI - Electrophoresis of concentrated mercury drops. AB - The electrophoretic behavior of concentrated monodispersed, positively charged mercury drops is investigated theoretically. The present study extends previous analyses by considering arbitrary surface potentials, double-layer polarization, and the interaction between adjacent double layers. The coupled equations describing the spatial variations in the flow field, the electric field, and the concentration field are solved by a pseudo-spectral method. For a low surface potential phi(r), the mobility increases monotonically with kappaalpha; kappa and alpha are respectively the reciprocal Debye length and the radius of a mercury drop. For medium and high phi(r), the mobility curve has a reflection point, which arises from the interaction of adjacent double layers, for kappaalpha. Also, if phi(r) is high, the mobility curve may exhibit a local minimum as kappaalpha varies. This phenomenon is pronounced if the concentration of the dispersed phase is high. If the double layer is thick, the mobility increases with phi(r), and the reverse is true if it is thin. We show that the higher the concentration of the dispersed phase the smaller the mobility, and as kappaalpha becomes large the mobility approaches a constant value, which is independent of the concentration of the dispersed phase. The mobility of mercury drops is larger than that of the corresponding rigid particles. PMID- 16256479 TI - Self-assembled film of poly(4-carboxyphenyl)acetylene and diazoresin and its photoelectric conversion property. AB - A novel multilayer film was assembled from water-soluble poly(4 carboxyphenyl)acetylene sodium salt (PCPA) and diazoresin (DR) in aqueous solution via electrostatic attraction. Under UV irradiation, following the decomposition of the diazonium group between the adjacent interfaces of the multilayers, the ionic bonds of the self-assembled film convert to covalent bonds and the film becomes very stable toward electrolyte aqueous solutions. Thus, the photoelectric conversion property of PCPA-containing film can be measured in a conventional three-electrode photoelectrochemical cell, and 0.5 mol/l KCl solution can be used as the supporting electrolyte. The photocurrent spectroscopy response coincides with the absorption spectrum of the irradiated self-assembled film, which indicates that the irradiated DR/PCPA film is responsible for the photocurrent generation. PMID- 16256478 TI - SnO2 thin films prepared by dip-coating from microwave synthesized colloidal suspensions. AB - Tin(IV) oxide thin films have been prepared by dip-coating. The suspensions used for these depositions have been synthesized by microwave-induced thermohydrolysis of tin tetrachloride aqueous solutions in the presence of hydrochloric acid. Single or multiple depositions were tested, on glass substrates as well as on pure SiO2. The obtained thin films were characterized by optical microscopy, interferometric roughness measurements (Micromap), scanning electron microscopy, secondary ion mass spectroscopy, and scanning tunneling microscopy. PMID- 16256480 TI - Delamination and intercalation of layered aluminophosphate with [Al3P4O16]3- stoichiometry in water/alcohol/amine solutions. AB - Delamination and intercalation of a layered microporous aluminophosphate [Al3P4O16](3-) .3[CH3CH2NH3]+ (AlP) were carried out in water/ethanol/amine and water/methanol/amine solutions. A good colloidal dispersion and saturated amine intercalates of the aluminophosphate can be obtained in solutions with dielectric constants in the range 50-70 and an amine concentration of 10 mmol/g AlP. The saturated C2-C12 amine intercalates of AlP with an even number of carbon atoms are packed with alkylamine bilayers tilted at an angle of 44.8 degrees in the interlayer region. PMID- 16256481 TI - Moving contact lines and Langmuir-Blodgett film deposition. AB - The objective of this paper is to point out the close relationship between contact line dynamics and LB film depositions, and it is designed to serve as a blueprint for future analysis of the LB technique. Moving contact lines and contact angles play a major role in Langmuir-Blodgett ultrathin film depositions. Although the effect of contact angles has been recognized for many years, a fundamental and comprehensive explanation of the phenomena taking place at the contact line has not been formulated before. Our understanding of contact line dynamics has improved thanks to careful experiments and new theoretical developments. Flow patterns depend on dynamic contact angle and the ratio of viscosities of the gas and liquid phases. More recently dynamic contact angles and flow patterns-have been linked to forces of molecular and double-layer origin. The dynamic relationship of flow patterns to interfacial and transport properties can be used to explain seemingly contradictory experimental results reported by researchers during more than 60 years of experience with the L-B technique. Windows of operability can be defined for X-type and Z-type depositions that are useful in the design of experimental and industrial L-B deposition equipment. PMID- 16256482 TI - Spreading of non-Newtonian liquids over solid substrates. AB - The spreading of drops of a non-Newtonian liquid (Ostwald-de Waele liquid) over horizontal solid substrates is theoretically investigated in the case of complete wetting and small dynamic contact angles. Both gravitational and capillary regimes of spreading are considered. The evolution equation deduced for the shape of the spreading drops has self-similar solutions, which allows obtaining spreading laws for both gravitational and capillary regimes of spreading. In the gravitational regime case of spreading the profile of the spreading drop is provided. PMID- 16256483 TI - Effect of multiple scattering on SANS spectra from bicontinuous microemulsions. AB - Small-angle neutron scattering is a powerful tool for investigating the microstructure of self-assembled systems. The domain length, d, and the correlation length, xi, are two measures of bicontinuous microemulsions that are often determined from coherent SANS spectra. Some microemulsions scatter strongly, however, so measured spectra can contain multiple coherent scattering, and neglect of that multiple scattering can lead to incorrect values of d and xi. In addition, multiple scattering can give rise to artifacts in the spectra, most notably an apparent scattering peak at twice the value of the scattering vector of the main peak. Here, changes in the SANS spectra from strongly scattering microemulsions and the parameters derived from them are reported as a function of relative scattering probability by varying both sample thickness and scattering contrast. A linear extrapolation of the results to zero scattering probability yields good estimates for the microstructural parameters, and the numerical procedure of Schelten and Schmatz is used to calculate the specific effects of multiple scattering on typical bicontinuous microemulsion scattering spectra. PMID- 16256484 TI - Emulsion stability based on phase behavior in sodium naphthenates containing systems: gels with a high organic solvent content. AB - Addition of heptane to a sodium naphthenates/toluene/water system at 25 degrees C reduces the lamellar liquid-crystal phase range and increases the microemulsion phase range. Both of these effects result in the extension of the composition range where emulsions have low stability. This effect is even stronger at 40 degrees C. Heptane addition also results in the formation of very stable emulsions within the overlapping phase-existence ranges of aqueous (L1) and organic (L2) phases. Stable non-birefringent gel observed in equilibrium with L1 and L2 phases contains only a small percentage of water and sodium naphthenates. The swelling behavior of an unstable gel, an emulsion previously compressed by centrifugation, appears to be due to a stepwise thickening of the thin liquid films between the droplets. PMID- 16256485 TI - Vesicle formation and other structures in aqueous dispersions of monoolein and sodium oleate. AB - The macroscopic appearance and microstructure in the dilute corner of the ternary monoolein (MO)-sodium oleate (NaO)-water (2H2O) system have been investigated by visual inspection and by using direct structural imaging with light microscopy and cryogenic transmission electron microscopy. The microstructural transformations that take place between the micellar phase (binary NaO2H2O axis) and the dispersed cubic phase (binary MO2H2O axis) upon increasing the ratio of MO to NaO are micelles, ruptured multilamellar vesicles together with flexible threads, various vesicle structures, vesicles in equilibrium with densely packed layers that either represent bilayers or domains of H(II) tubes visible from the side, and finally a pattern that may be either a cubic phase or domains of H(II) tubes visible in a cross section. Spontaneously formed uni- and multilamellar vesicles that show long-term stability are found to be the dominant structure for mixed dispersions over almost the entire concentration range. The addition of NaOH to the non-bilayer-forming system, the ternary MOOA (oleic acid)-2H2O system, leads to the formation of vesicles. Vesicles were also observed in other ternary MO-aqueous-based systems with potassium oleate, cetyltrimethylammonium bromide, sodium taurodeoxycholate, or dipalmitoylphosphatidylcholine instead of NaO. PMID- 16256486 TI - Synergism in mixtures of cationic surfactant and anionic copolymers. AB - Surface tension measurements have been made in aqueous solutions of anionic hemiesters of an alternating copolymer of maleic acid and styrene, MAS-n with n=0 12, in the presence of dodecyltrimethylammonium bromide, DTAB. A synergistic aspect of surface tension reduction efficiency was observed for all systems studied. The pseudo-phase separation approach and regular solution approximation have been applied, and the interaction parameter, beta, and the mole fraction of DTAB in the adsorbed layer (on a surfactant/repetitive unit basis), X, were obtained. Negative values of beta, ranging from -3 to -11, were calculated. On the other hand, the molar fraction of DTAB varies from 0.52 to 0.26. These results are discussed in terms of hydrophobic effects on the distribution of the aggregates between the interface and the bulk of the solution. The conditions predicted by the model to obtain synergism in the tension reduction efficiency are completely satisfied in all cases. PMID- 16256487 TI - Miscibility of sodium chloride and decyl methyl sulfoxide in the adsorbed film and micelle. AB - The surface tension of aqueous solutions of a sodium chloride (NaCl)-decyl methyl sulfoxide (DeMS) mixture was measured as a function of the total molality of the mixture and the mole fraction of DeMS in the mixture at 298.15 K under atmospheric pressure. The total surface density of the mixture and the mole fraction of DeMS in the adsorbed film and micelle were numerically evaluated by applying the thermodynamic treatment of surfactant mixture to the NaCl-DeMS mixture. Miscibility of NaCl and DeMS in the adsorbed film and micelle was clarified by use of the phase diagram of adsorption and micelle formation. Positive adsorption of NaCl was observed in the presence of DeMS and attributed to attractive interaction between the polar head group of DeMS molecule and Na+ or Cl- ions in the adsorbed film and micelle. The results were compared with those of NaCl-octyl methyl sulfoxide and NaCl-decyldimethylphosphine oxide mixtures to elucidate the structure effect of nonionic surfactant on the miscibility. PMID- 16256488 TI - Binary phase equilibria of three alpha-methyl-substituted sodium alkanoate surfactants. AB - In this work, we continue our study of methyl-substituted surfactants and present the aqueous binary phase diagrams of racemic sodium 2-methyloctanoate, nonanoate, and -dodecanoate, respectively. All systems have very low Krafft temperatures within the solution phase, between 1 and 4 degrees C. The phase sequences of the two shorter surfactants are very similar to those of the unsubstituted sodium octanoate, although with somewhat different range of existence for the phases formed. The sodium 2-methyldodecanoate system is different from the unsubstituted sodium dodecanoate system, as the former seems to lack a hexagonal phase. The surfactant systems were delineated using 2H NMR splittings and crossed polarizers, and combined with SAXS for determination of phase structure. PMID- 16256489 TI - The role of the surfactant head group in the emulsification process: Single surfactant systems. AB - To clarify the effect of the surfactant head group on the emulsification process, dilute dodecane in water emulsions were prepared in a small flow-through cell with three surfactants which had the same hydrocarbon tail length but different head groups. The different surfactants types were (a) a nonionic, hexa(ethyleneglycol) mono n-dodecyl ether (C12E6), (b) an anionic, sodium dodecyl sulfate (SDS), and (c) a cationic, n-dodecyl pyridinium chloride (DPC), and the emulsions were prepared under the same conditions. From dynamic light scattering measurements, it was shown that the mean steady state droplet size of the emulsions (obtained after 20 min dispersion) could be related to the interfacial tension at concentrations in the region of the cmc. This result was in agreement with laminar and turbulent viscous flow theory. However, the particle size versus surface tension data for the different surfactant systems did not fall on a single line. This behavior suggested that the surfactant played a secondary role in defining the droplet size (in addition to reducing the interfacial tension) possibly through diffusion and relaxation, during deformation of the interface. In addition, it was found that the values of the equilibrium "surfactant packing densities" of the different surfactants at the oil/water interface were almost equal near the cmc, but the mean droplet size and the interfacial tension at the cmc decreased following the order DPC>SDS>C12E6 . PMID- 16256490 TI - Morphologies of three-phase emulsions of the ternary nonionic amphiphile/oil/water systems and their determination by electrical method. AB - Equations have been developed for the electrical and thermal conductivities of dispersions of two different phases of low conductivity in a third, conductive phase. These equations predict the conductivity of the dispersion from the volume fractions and conductivities of the constituent phases. Electrical conductivity measurements on dispersions of three liquid phases, i.e., three-phase emulsions, were made over a wide range of volume fractions of each dispersed phase and used to test the equations. The equations predict accurately dispersion conductivities from the measured volume fractions and conductivities of the constituent phases without any adjustable parameters. The predicted values are in excellent agreement with the measured conductivities of three-phase emulsions in the nonionic amphiphile/oil/water systems. This leads to the determination of three phase emulsion morphologies. When the ratio Kd of the emulsion-drop conductivity to the continuous phase conductivity is O(10(-1)) 12. In addition, the resultant nickel nanoparticles could be magnetically recovered and re-dispersed in ethylene glycol without size change and agglomeration. The magnetic measurements indicated they were superparamagnetic with a saturation magnetization of 22 emu/g, a remanent magnetization of 6.4 emu/g, and a coercivity of 0.1 Oe at a mean diameter of 9.2 nm. Also, the magnetization increased with decreasing temperature due to the decrease in thermal energy. All the observed magnetic properties essentially reflected the nanoparticle nature. Furthermore, it was found that hydrazine was catalytically decomposed to hydrogen and nitrogen gases by the resultant nickel nanoparticles. The corresponding decomposition rate at 25 degrees C and 1 atm was 3.1 nmol/(h mg of Ni). PMID- 16256508 TI - Kinetics of dissolution of powdered Pentelic marble in under-saturated solutions: the role of particle characteristics. AB - In the present study, the effect of mesopore and particle size distributions on the kinetics of dissolution of powdered Pentelic marble was investigated. Powders obtained by grinding marble slabs in an agate mortar with a pestle (1-3.5 min) were found to be non-porous by nitrogen absorption measurements. These powders, upon dissolution under conditions of constant under-saturation, 25 degrees C, pH 8.25, showed that the kinetics in the absence of mesopores depended on the number of active sites on the exposed surface. Thus, powders consisting of smaller particles, having higher specific surface areas, yielded higher rates of dissolution. Powders, however, which were prepared by grinding marble slabs in a cylinder mill for time periods between 2.0 and 30.0 min, and which exhibited considerable mesoporosity, showed the opposite trend. The rates of dissolution measured for these powders in under-saturated solutions increased with increasing mean particle size and decreased with increasing specific surface area. This finding suggested that the presence of mesopores resulted in lower dissolution rates even though the exposed total surface area was larger. Furthermore, the larger the number of mesopores in a powder sample the slower the corresponding dissolution rates in under-saturated solutions. PMID- 16256509 TI - Fouling mitigation of anion exchange membrane by zeta potential control. AB - The feasibility of fouling mitigation of anion exchange membranes (AEMs) in the presence of humate was studied by adding three different types of water-soluble polymers, i.e., poly(acrylic acid) (PAA), poly(vinyl alcohol) (PVA), and poly(ethylene imine) (PEI), during electrodialysis (ED) desalination. Measurement of zeta potential of the humate used in this study showed highly negative potential (about -30 mV), implying that the humate had a strong fouling potential on the AEMs in ED. Of the three water-soluble polymers, PEI showed a positive zeta potential (about +14 mV) and is able to form an interpolymer complex with the humate. PAA and PVA hardly formed interpolymer complexes with humate due to electrostatic repulsion. The PEI-humate mixture with a volume ratio of 1:20 (PEI:humate) showed zero zeta potential, and a complexed humate with zero surface charge was formed, resulting in no fouling effects on the AEMs. Accordingly, the desalting ED experiments with PEI showed improved ED performance. Further, black colloids formed in the mixture did not cause the cell resistance to increase. PMID- 16256510 TI - Drag force on a porous, non-homogeneous spheroidal floc in a uniform flow field. AB - The force acting on a porous spheroidal floc having a nonhomogeneous structure in a uniform flow field is evaluated theoretically. Here, the floc is simulated by an entity having a two-layer type of structure, and its porous nature is mimicked by varying the relative magnitudes of the permeabilities of its inner and outer layers. The results of numerical simulation reveal that, for the same volume averaged permeability, the drag coefficient of a spheroidal floc with a nonhomogeneous structure is much larger than that of a floc with a homogeneous structure for both prolate and oblate spheroids. This is true regardless of the relative magnitudes of the permeability of the inner layer and that of the outer layer. While the drag coefficient of a homogeneous prolate is the same as that of a homogeneous oblate the drag coefficient of a nonhomogeneous prolate is larger than that of a nonhomogeneous oblate. For the same volume-averaged size, the more nonhomogeneous the structure of a spheroidal floc the easier for the relation between the drag coefficient and the Reynolds number to deviate from a Stokes-law like relation. For a fixed volume-averaged permeability, the effective drag coefficient increases with the increase in the ratio (polar radius of inner layer/polar radius of floc), regardless of whether its inner layer is less permeable than its outer layer or not. PMID- 16256511 TI - Nonlinear Rayleigh-Taylor instability for hydromagnetic Darcian flow: effect of free surface currents. AB - Capillary-gravity waves of permanent form at the interface between two unbounded magnetic fluids in porous media are investigated. The system is influenced by the horizontal direction of the magnetic field to the separation face of two semi infinite homogeneous and incompressible fluids, so that the fields allow free surface currents at the interface. The solutions of the linearized equations of motion under nonlinear boundary conditions lead to derivation of a nonlinear equation governing the interfacial displacement. This equation is accomplished by using the cubic nonlinearity. Taylor theory is used to expand the governing nonlinear equation in the light of the multiple scales in both space and time. The perturbation analysis leads to imposition of two levels of solvability conditions, which are used to construct the well-known nonlinear Ginzburg-Landau equation. The stability criteria are discussed theoretically and numerically and stability diagrams are obtained. Regions of stability and instability are identified for the surface current density. It is found that the stabilizing role for the magnetic field is retarded when the flow is in porous media. Moreover, the increase in the values of resistance parameters plays a dual role, in stability behavior and in the increase in surface current density. PMID- 16256512 TI - Correlation of structural and permeation properties in sol-gel-made nanoporous membranes. AB - The aim of the present work was to establish a fundamental link between the basic structural properties of ceramic nanoporous membranes made by the sol-gel process and their respective transport properties, for a systematic evaluation of their performance in gas and liquid applications. For this purpose, supported and unsupported gamma-Al2O3 and TiO2 membranes were prepared from different colloidal dispersions (sols) by a sol-gel dipping process followed by drying and calcination, resulting in structures of crystallites of different shape and stacking arrangement. Accordingly, the pore structure of each membrane was simulated employing process-based reconstruction techniques and the permeation properties were predicted by solving the appropriate transport equations in the generated structures. Excellent agreement was achieved between the computed and experimental permeability values in the Knudsen and viscous flow regimes, validating the considerations made regarding the basic structural characteristics and the procedure for generation of the membrane structures. Moreover, it was shown that the shape and stacking arrangement of the primary particles (crystallites) of the sol have a major impact on the formation of pathways in the membrane pore structures and control the transport and therefore also the separation properties of these materials. PMID- 16256513 TI - Comparison of the structural properties of isomorphously substituted Fe in mordenite zeolites prepared by different methods. AB - Fe was introduced in mordenite zeolite by means of ion exchange either in solid or in liquid state. The iron loading (50--200 wt%), iron precursor (FeSO4.7H2O and FeCl3), and mordenite starting material (NH4M, HM, and NaM) were varied during the exchange processes. The Fe species were characterized by N2 adsorption measurements as well as by XRD and Mossbauer spectroscopies. The Fe-mordenite samples prepared by liquid-state ion exchange attained remarkable Fe dispersion and surface areas higher than those of the parent. It was found that Fe3+ ions, which substituted the framework Al and accordingly occupied tetrahedral sites, were decreased with Fe loadings with concomitant increase in Fe3+-occupied octahedral sites. The latter sites disappeared at 20 K to provoke the superparamagnetic alpha-Fe2O3 in different particles size. The acid leaching (0.1 M HCl, 333 K, 3 h) of the samples showed the disappearance of the most highly distorted extra-framework Fe3+ species, providing an indication of their presence on the external surface. On the other hand, a hematite phase was detected in the solid-state ion exchange of FeCl3 with either HM or NH4M at the loading of 100% Fe. More correlations between Mossbauer data on one hand and XRD and texturing properties on the other hand were evaluated and discussed. PMID- 16256514 TI - Channel flow cell studies on the evaluation of surface pretreatments using phosphoric acid or polymaleic acid for calcite stone protection. AB - The dissolution kinetics of surface-pretreated and weathered calcite was investigated in dilute acid using a channel flow cell with microdisk detection. Two pretreatments were studied, polymaleic acid and phosphoric acid. Treatment with polymaleic acid was shown to significantly passivate calcite but to a lesser extent than the phosphoric acid and the former coating was found to be less effective for protection of calcite from acid attack. However, treatment of calcite with phosphoric acid resulted in the passivation of calcite from acid attack which strongly inhibited dissolution, an effect that was enhanced even further after exposure to the environment. PMID- 16256515 TI - Quasistatic detachment of a sphere from a liquid interface. AB - In this paper the problem of removing a spherical particle initially attached to a liquid-gas interface is analytically treated. In particular, the Derjaguin equation for small radii is used to derive a closed-form approximate expression for the work of detachment of the sphere from the interface. Expressions corresponding to the prescribed displacement condition and the applied force condition, which seems to be the relevant condition for the flotation separation process, are presented. A special effort has been made to closely compare analytical results with data obtained through the exact numerical treatment of the detachment process. Results show that proposed expressions are sufficiently accurate to calculate the energy required to detach the sphere from the interface as soon as the sphere radius is small compared to the capillary length. Validity limits are specified. PMID- 16256516 TI - Experimental investigation of contact angle, curvature, and contact line motion in dropwise condensation and evaporation. AB - Image-analyzing interferometry is used to measure the apparent contact angle and the curvature of a drop and a meniscus during condensation and evaporation processes in a constrained vapor bubble (CVB) cell. The apparent contact angle is found to be a function of the interfacial mass flux. The interfacial velocity for the drop during condensation and evaporation is a function of the apparent contact angle and the rate of change of radius of curvature. The dependence of velocity on the apparent contact angle is consistent with Tanner's scaling equation. The results support the hypothesis that evaporation/condensation is an important factor in contact line motion. The main purpose of this article is to present the experimental technique and the data. The equilibrium contact angle for the drop is found experimentally to be higher than that for the corner meniscus. The contact angle is a function of the stress field in the fluid. The equilibrium contact angle is related to the thickness of the thin adsorbed film in the microscopic region and depends on the characteristics of the microscopic region. The excess interfacial free energy and temperature jump were used to calculate the equilibrium thickness of the thin adsorbed film in the microscopic region. PMID- 16256517 TI - Viscoelastic behavior of surfactant threadlike micellar solutions: effects of additives, 4. AB - Viscoelastic behavior of aqueous solutions of a cationic surfactant, cetyltrimethylammonium p-toluene sulfonate (CTApTS), and an organic salt, sodium p-toluene sulfonate (NapTS), was examined at room temperature by changing concentrations of CTApTS (CD) and NapTS (CS). CTApTS forms long threadlike micelles in aqueous solution even at low CD in spite of the absence of additives. Aqueous CTApTS solutions show pronounced viscoelastic behavior, which is perfectly described by Maxwell model type viscoelasticity, keeping a single set of relaxation parameters (GN and tau). The magnitude of relaxation strength, GN, is proportional to the square of CD irrespective of the value of CS. The value of a relaxation time, tau, is independent of CD, while the value remarkably reduces with increasing concentration (CS*) of excess pTS- ions in the bulk aqueous phase. The relationship between tau and CS* for the system is not identical with that for aqueous solutions of cetyltrimethylammonium bromide (CTAB) and NapTS at CS* < 30 mmolkg(-1). However, the relationships between tau and CS* for these systems agree well with each other at CS* >/= 30 mmolkg(-1). This implies that formed threadlike micelles in both systems have similar structure and the relaxation mechanism is controlled by CS*, but not influenced by the presence of Br- ions above CS* = 30 mmolkg(-1). PMID- 16256518 TI - Behavior of soybean oil-in-water emulsion stabilized by nonionic surfactant. AB - A soybean oil-in-water emulsion was prepared using nonionic Tween series surfactants. The effects of temperature, hydrophilic-lipophilic balance (HLB) value of the surfactant, and surfactant-to-oil ratio on the size of emulsion drops were investigated with an acoustic and electroacoustic devices. In the case of Tween 85, the influence of pH and electrolyte on the zeta potential of emulsion drops was examined. zeta potential appears to be strongly dependent on pH, varying in the case of high concentrations of NaCl from +60 to -90 mV. High concentrations of Na+ and K+ are capable of separating the oil phase from the emulsion. Divalent cations such as Ca2+ and Mg2+ lead to two points of zero charge at high concentrations, which is not observed for the case of hydrolyzable trivalent species like Al3+ and Fe3+. PMID- 16256519 TI - Microstructures in the aqueous solutions of a hybrid anionic fluorocarbon/hydrocarbon surfactant. AB - The aqueous solutions of the anionic hybrid fluorocarbon/hydrocarbon surfactant sodium 1-oxo-1[4-(tridecafluorohexyl)phenyl]-2-hexanesulfate (FC6HC4) shows peculiar rheological behavior. At 25 degrees C the viscosity vs concentration curve goes successively through a maximum and a minimum, while the viscosity vs temperature curve of the 10 wt% aqueous FC6HC4 solution goes through a marked maximum at 36 degrees C [Tobita et al., Langmuir 13 (1997) 5054]. In an attempt to explain these properties the microstructure of aqueous solutions of FC6HC4 has been investigated by means of digital light microscopy, transmission electron microscopy at cryogenic temperature (cryo-TEM), rheology, and self-diffusion NMR. At 20 degrees C, the increase of the FC6HC4 concentration was found to result in a progressive change of structure of the surfactant assemblies from mainly spherical micelles at 0.5 wt% to mainly cylindrical micelles at 10 wt%. At intermediate concentrations small disk-like micelles and small complete and incomplete vesicles coexisting with cylindrical micelles were visualized. The occurrence of stretched cylindrical micelles is responsible for the effect of the surfactant concentration on the solution viscosity. Cryo-TEM, rheology, and self diffusion NMR all suggest that an increase of the temperature brings about a growth of the assemblies present in the 10 wt% solution of FC6HC4. The structure of the assemblies present at the temperature where the viscosity is a maximum could not be elucidated by cryo-TEM because of the probable occurrence of an on the-grid phase transformation, the result of blotting during specimen preparation. Nevertheless, the results show that the observed large assemblies break up at higher temperature to give rise to a more labile bicontinuous structure that consists of multi-connected disordered lamellae, with many folds and creases, and that may well be the L3 phase. PMID- 16256520 TI - Effect of solvent composition on dispersing ability of reaction sialon suspensions. AB - This work focuses on the optimization of the rheological behavior of suspensions considering different solvent compositions. The effects of methyl ethyl ketone (MEK)/ethanol (E) solvent mixtures on reaction sialon suspensions were investigated by measuring sedimentation behavior, adsorption of dispersant, and flow behavior. It was shown that both the flow behavior and the sedimentation behavior strongly depended on selection of solvent composition. Using 3 wt% KD1 as dispersant, well-dispersed colloidal suspensions could be obtained in MEK-rich solvents. The suspensions with 60 vol% MEK/40 vol% E as solvent could be fitted to the Bingham model with very low yield stress, while suspensions with pure MEK or ethanol-rich mixtures as solvent showed pseudo plastic behavior with relatively high yield stress values. A model was proposed to explain the different flow behaviors of suspensions considering the different configurations of dispersant at particles' surfaces. PMID- 16256521 TI - Interfacial properties of adsorbed films made of a PEG2000 and PLA50 mixture or a copolymer at the dichloromethane-water interface. AB - Adsorption kinetics of films of poly(ethylene glycol) (PEG2000) studied by the dynamic pendant drop method showed that PEG2000 was more tensioactive at the dichloromethane (DCM)-water interface than at the air-water interface. When initially solubilized into DCM, PEG2000 segments would form an adsorbed layer with hydrophobic segments buried into the polymer chains turned toward the organic phase. Compression of this layer, accompanied by viscoelastic effects, led to expulsion of some hydrophilic tails toward the water phase. When initially dissolved in water, adsorption of PEG2000 segments led to an elastic PEG2000 layer organized on both sides of the interface. Results showed that when the PEG2000-PLA50 (poly(D,L-lactide)) copolymer film was adsorbed at the DCM-water interface, it resulted in a mixed layer exclusively turned toward DCM and its rheological properties were governed by PLA50. When adsorption at the DCM-water interface resulted from a physical mixture of PEG2000 and PLA50, rheological properties of the film were influenced by the initial localization of PEG2000 in the bulk phases. In the case of a mixed film formed by the adsorption of PLA50 from DCM and PEG2000 from water, results showed that PEG2000 segments totally pushed those of PLA50 away from the interface and exclusively influenced the behavior of the mixed film. PMID- 16256522 TI - Heavy metal immobilization in aqueous solution using calcium phosphate and calcium hydrogen phosphates. AB - This study examines the possibilities for removing heavy metal cations from water with calcium phosphate, calcium hydrogen phosphate, and calcium dihydrogen phosphate at 293 K. It was reported that immobilization of aqueous heavy metal cations, which is known to be one of the characteristic properties of calcium hydroxyapatite, proceeded favorably with these phosphates. Calcium phosphate, calcium hydrogen phosphate, and calcium dihydrogen phosphate could favorably remove Pb2+ from aqueous solution. Calcium hydrogen phosphate also removed aqueous Cu2+, Co2+, and Cd2+, whereas these cations were not immobilized by calcium phosphate and calcium dihydrogen phosphate. A contribution of the dissolution-precipitation mechanism to immobilization with these phosphates is suggested. PMID- 16256523 TI - Synthesis of one-dimensional magnetic Co nanoparticles in a novel solution system. AB - Magnetic Co nanoparticles with different morphologies were synthesized in a novel solution system using a UV irradiation technique. By adjusting the compositions in the solution, long nanowires with different aspect ratios as well as spherical nanoparticles with controllable particle size could be obtained. PMID- 16256524 TI - Influence of heat treatment of rayon-based activated carbon fibers on the adsorption of formaldehyde. AB - The influence of heat treatment of rayon-based activated carbon fibers on the adsorption behavior of formaldehyde was studied. Heat treatment in an inert atmosphere of nitrogen for rayon-based activated carbon fibers (ACFs) resulted in a significant increase in the adsorption capacities and prolongation of breakthrough time on removing of formaldehyde. The effect of different heat treatment conditions on the adsorption characteristics was investigated. The porous structure parameters of the samples under study were investigated using nitrogen adsorption at the low temperature 77.4 K. The pore size distributions of the samples under study were calculated by density functional theory. With the aid of these analyses, the relationship between structure and adsorption properties of rayon-based ACFs for removing formaldehyde was revealed. Improvement of their performance in terms of adsorption selectivity and adsorption rate for formaldehyde were achieved by heat post-treatment in an inert atmosphere of nitrogen. PMID- 16256525 TI - Physicochemical characterization and adsorption behavior of calcined Zn/Al hydrotalcite-like compound (HTlc) towards removal of fluoride from aqueous solution. AB - A Zn/Al hydrotalcite-like compound (HTlc) was prepared by co-precipitation (at constant pH) method and was characterized by XRD, TG/DTA, FTIR, and BET surface area. The ability of Zn/Al oxide to remove F- from aqueous solution was investigated. All the adsorption experiments were carried out as a function of time, pH, concentration of adsorbate, adsorbent dose, temperature etc. It was found that the maximum adsorption takes place within 4 h at pH 6.0. The percentage of adsorption increases with increase in the adsorbent dose, but decreases with increase in the adsorbate concentration. From the temperature variation it was found that the percentage of adsorption decreases with increase in temperature, which shows that the adsorption process is exothermic in nature. The adsorption data fitted well into the linearly transformed Langmuir equation. Sulfate and phosphate were found to have profound effects on fluoride removal. Thermodynamic parameters such as DeltaG0, DeltaH0, and DeltaS0 were calculated. The negative value of DeltaH0 indicates that the adsorption process is exothermic. The apparent equilibrium constants (Ka) are also calculated and found to decrease with increase in temperature. With 0.01 M NaOH the adsorbed F- could be completely desorbed from Zn/Al oxide in 6 h. PMID- 16256526 TI - Sorption of uranium (VI) species on zircon: structural investigation of the solid/solution interface. AB - This work is an investigation of the mechanisms of interaction between uranium (VI) ions and zirconium silicate. The speciation of uranium (VI) sorbed on zircon was studied using four complementary techniques as probes of the local structure around the uranium atom: laser spectrofluorimetry, X-ray photoelectron spectroscopy (XPS), diffuse reflectance infrared Fourier-transformed (DRIFT) spectroscopy, and EXAFS spectroscopy. The sorption of uranyl on zirconium oxide was also studied to allow structural comparisons. Spectrofluorimetry and XPS results allowed an identification of the silicate sorption sites on the solid. These methods associated with spectrofluorimetry and DRIFT led to a characterization of the sorbed surface complexes, taking into account the influence of the nature of the background salt and of the pH on the structure of the U(VI) surface species. EXAFS measurements, either on air-dried samples or in situ, were then carried out on well-characterized samples and allowed identification of the sorption mechanism on zircon as the formation of an inner sphere polydentate surface complex. PMID- 16256527 TI - Competitive adsorption of surfactants at air/water interfaces. AB - The zeta potential of an air bubble suspended in an aqueous solution of mixed fluorocarbon and hydrocarbon anionic surfactants is studied over a wide range of concentrations and mixture compositions. The zeta potential is related to surfactant ion adsorption. The two surfactants, which exhibit an antipathy manifested by micellar demixing, compete for surface sites. The total surfactant adsorption is reduced when both surfactants are present. Adsorption phenomena are closely correlated to the micellar phase diagram. PMID- 16256528 TI - Preparation and characterization of activated carbon fibers supported with silver metal for antibacterial behavior. AB - In this work, the effect of immersion in silver nitrate solution on activated carbon fibers (ACFs) was investigated in relation to adsorption behavior and antibacterial activity of ACFs supported with silver (ACF/Ag). The pore and surface properties were studied in terms of BET volumetric measurement with nitrogen adsorption, X-ray diffraction (XRD), and scanning electron microscopy (SEM). The antibacterial activities of ACF/Ag were studied in broth dilution tests against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) from a point of view of water purification. As an experimental result, the silver content of ACF/Ag increased with increasing concentration of silver nitrate. It was found that the micropore structure slightly decreased as the silver nitrate concentration increased. Otherwise, it was revealed that the ACF/Ag possessed a strong antibacterial activity and an inhibitory effect for the growing of E. coli and S. aureus, respectively. Silver content on ACF/Ag decreased rapidly because of rough morphology of silver particles in water erosion. PMID- 16256529 TI - Morphology and surface heterogeneities in synthetic goethites. AB - In the framework on a study of the acido-basic and sorption properties of iron oxides, a thorough characterization of two types of goethite powders was performed in several laboratories joined in a common project. Chemical analysis by ICPAES; high-resolution SEM, TEM, and AFM observations; XRD with line width analysis; and argon and nitrogen sorption isotherms were used for that purpose. The main crystallographic faces of goethite particles could be identified as {001}, {101}, and {121}, and their abundance correlated with the distribution of low-pressure argon adsorption local isotherms. These results will be very useful for further studies on the relationship between surface reactivity in aqueous solution and orientation of solid surfaces. PMID- 16256530 TI - Asymmetric alpha-helicity loss within a peptide adsorbed onto charged colloidal substrates. AB - A combination of circular dichroism and solution 1H NMR spectroscopy provides a localized description of the distribution of alpha -helical structure within the capped peptide DDDDAAAAARRRR (4DAR5) in aqueous solution and adsorbed onto anionic and cationic colloidal substrates. The adsorption-induced conformational changes are different from those observed upon heating 4DAR5 in solution, in which case the alanine segment remains largely alpha -helical and the transition to a coil structure propagates from the termini. Adsorption is driven by electrostatic complementarity, which places the charged peptide segment adjacent to the substrate of opposite charge. A similar pattern of alpha -helicity loss is observed whether the peptide is adsorbed onto anionic or cationic colloidal silica, despite inverse orientations; significant alpha -helicity loss occurs within the central alanine segment and the terminal arginine segment, whereas alpha -helicity is retained in the aspartate segment. This pattern of adsorption induced conformational change illustrates the complex and subtle balance among the intramolecular and intermolecular factors that influence the conformations of adsorbed peptides and proteins. PMID- 16256531 TI - Alumina interaction with AMPS-MPEG random copolymers I. Adsorption and electrokinetic behavior. AB - Adsorption of brush copolymers, bearing sulfonate groups and polyethylene glycol segments, on to alumina particles in suspension in water has been investigated. Study of the adsorption isotherms revealed that the copolymers displayed a strong affinity for the surface of the alumina regardless of the fraction of ionic groups on the polymer. For poly(ethylene glycol) content greater than 50%, the adsorption isotherms revealed an initial adsorption plateau followed by a second one. The shape of the adsorption isotherms was interpreted in terms of the polymer configuration at the solid-to-liquid interface. The effects of the pH and the ionic force on adsorption were studied and connected to the effects of interaction between chain segments at the surface of the alumina particles. Changes in the electrokinetic properties of the alumina particles after addition of the copolymers were investigated by following the zeta potential of particles as a function of pH. In the presence of the copolymer continuous shift of the isoelectric point IEP to a more acidic values was observed. Beyond a certain concentration the zeta potential remained negative regardless of the pH. PMID- 16256532 TI - Surfactant displacement of human serum albumin adsorbed on loosely packed self assembled monolayers: cetyltrimethylammonium bromide versus sodium dodecyl sulfate. AB - The surfactants sodium dodecyl sulfate (SDS) and cetyltrimethylammonium bromide (CTAB) displace human serum albumin (HSA) from loosely packed self-assembled monolayers (SAM) of hydrophobic alkyl chains by different means. Removal of HSA is of interest because previous work has suggested that the adsorption of HSA to such loosely packed SAMs may be sufficiently tenacious to offer opportunities for surface passivation. While HSA remains on the surface after exposure to SDS and rinsing, no protein remains after exposure to CTAB and rinsing. X-ray reflectivity and X-ray photoelectron spectroscopy measurements indicate that CTAB molecules remain interdigitated in the loosely packed SAM after rinsing, suggesting that CTAB is more effective in removing the HSA because it interacts more strongly with the SAM. PMID- 16256533 TI - Microbial surface thermodynamics and interactions in aqueous media. AB - Microbial surface thermodynamics and interactions in aqueous media were investigated for seven typical rod-shaped bacterial strains of Enterobacteriaceae, Pseudomonadaceas, and Bacillaceae, which included Escherichia coli HB101, Escherichia coli JM109, Pseudomonas fluorescens, Pseudomonas putida, Pseudomonas aeruginosa, Pseudomonas sp., and Bacillus subtilis. All the microorganisms studied exhibited a monopolar and predominant hydrophilic surface and a negative Delta G132tot, total free energy of interactions between microbes (1) and silica gel or Canadian River Alluvium (CRA) (2) immersed in water (3) at the equilibrium distance, which accounted for their attachment on the medium surface. The microbial attachment was proportional to the corresponding Delta G132tot values. Among the microorganisms studied, B. subtilis had the most attachment on both silica gel and CRA because it had the smallest Delta G132tot values (-17.14kT for silica gel and -21.84kT for CRA). The origins of Lifshitz van der Waals, Lewis acid/base, and electrostatic interactions were discussed and related to experimental observations. PMID- 16256534 TI - Study of adsorption and viscoelastic properties of proteins with a quartz crystal microbalance by measuring the oscillation amplitude. AB - The adsorption kinetics of protein A, BSA, IgG, and fibronectin has been investigated using a homemade quartz crystal microbalance. Information about the energy losses appearing in the system is measured by the maximal oscillation amplitude and the dissipation factor. Only the maximal oscillation amplitude allows us to distinguish the different contributions of liquid and mass to the total frequency shift. The adsorption of proteins has been performed on Ti and Au surfaces at different concentrations. The amount of irreversible adsorbed protein A and IgG increases with increasing bulk concentrations. On Au more proteins adsorb, but their biological activity is reduced in comparison to Ti. Protein A forms a first monolayer in a few seconds, which shows practically no energy losses, and following this a second monolayer is formed. The adsorption rate for the second monolayer is much smaller and energy losses are present. Fibronectin is forming a very viscoelastic system, whose mechanical properties are affected by immersion in different buffer solutions. PMID- 16256535 TI - Adsorption of human serum albumin (HSA) onto colloidal TiO2 particles, Part I. AB - The adsorption of human serum albumin (HSA) onto colloidal TiO2 (P25 Degussa) particles was studied in NaCl electrolyte at different solution pH and ionic strength. The HSA-TiO2 interactions were studied using adsorption isotherms and the electrokinetic properties of HSA-covered TiO2 particles were monitored by electrophoretic mobility measurements. The adsorption behavior shows a remarkable dependence of the maximum coverage degree on pH and was almost independent of the ionic strength. Other characteristic features such as maximum adsorption values at the protein isoelectric point (IEP approximately 4.7) and low-affinity isotherms that showed surface saturation even under unfavorable electrostatic conditions (at pH values far away from the HSA IEP and TiO2 PZC) were observed. Structural and electrostatic effects can explain the diminution of HSA adsorption under these conditions, assuming that protein molecules behave as soft particles. Adsorption reactions are discussed, taking into account acid-base functional groups of the protein and the surface oxide in different pH ranges, considering various types of interactions. PMID- 16256536 TI - Investigation of protein adsorption and electrochemical behavior at a gold electrode. AB - The adsorption of two model proteins, human serum albumin and immunoglobulin G, on a gold electrode surface was investigated using 125I radiolabeling and cyclic voltammetry (CV). 125I radiolabeling was used to determine the extent of protein adsorption, while CV was used to ascertain the effect of the adsorbed protein layer on the electron transfer between the gold electrode and an electroactive moiety in solution, namely, K3Fe(CN)6. The adsorbed amounts of HSA and IgG agreed well with previous results and showed approximately monolayer coverage. The amount of adsorbed protein increased when a positive potential (700 mV) was applied to the electrode, while the application of a negative potential (-800 mV) resulted in a decrease. When the solution pH was varied to alter the charge on the protein, the adsorption trends appeared to follow electrostatic interaction, namely, greater adsorption when the electrode and the protein possessed opposite charge and vice versa. The adsorbed protein layer had the effect of blocking the electron transfer. It was possible to correlate the degree of electron blocking with the amount of adsorbed protein to show that the greater the adsorption, the larger the blocking effect. Of the two proteins used, HSA proved to be more efficient at blocking the electron transfer. PMID- 16256537 TI - Solid-state 1H and 27Al NMR studies of amorphous aluminum hydroxides. AB - Two kinds of amorphous aluminum hydroxides, a sample precipitated from admixing AlCl3 and NaOH aqueous solutions and the commercial product, were measured by 27Al and 1H solid-state NMR spectroscopy. Pentahedral and tetrahedral coordinations, as well as octahedral coordination of oxygen atoms for aluminum, are observed in 27Al magic angle spinning (MAS) spectra of both amorphous samples. In contrast, octahedral coordination is only observed in gibbsite, bayerite, and boehmite. According to 1H MAS-NMR spectra under conditions of high spinning rate (35 kHz) and high field (14.09 T), free waters and OH groups coupled with aluminum for amorphous samples are observed at approximately 5 and approximately 4.5 ppm, respectively, the latter peak being broader. This is consistent with the differential spectra between spin echo and transfer of populations in double resonance. We conclude that the subunits of AlO4, AlO5, and AlO6 in amorphous aluminum hydroxides are bound through hydrogen bonds with a wide distribution of bonding strength. PMID- 16256538 TI - Preparation of nearly monodisperse multiply coated submicrospheres with a high refractive index. AB - Nearly monodisperse SiO2/TiO2/SiO2 multiply coated submicrospheres with nearly monodisperse silica submicrospheres as cores, thick titania layers, and thin silica skin were prepared to increase the refractive index of complex submicrospheres while keeping their near monodispersity and perfect surface properties. Nearly monodisperse colloidal silica submicrospheres as cores with a diameter of 200 nm were synthesized by a seeding technique on the basis of the hydrolysis of tetraethyl orthosilicate (TEOS) in an aqueous ethanol solution with ammonia as catalyst. On the basis of the hydrolysis of tetrabutyl orthotitanate, a procedure combining continuous feeding with multistep coating was determined to prepare titania coatings about 40 nm thick and increase the refractive index of the complex submicrospheres. The hydrolysis of TEOS was still used to get the outmost silica coating about 10 nm thick on titania coated silica submicrospheres to eliminate random aggregation caused by the TiO2 surface properties of the TiO2/SiO2 complex submicrospheres during the final fabrication of photonic crystals. PMID- 16256539 TI - Layer-by-layer self-assembly of Prussian blue colloids. AB - The adsorption of Prussian blue (PB) colloids within layers of polyelectrolytes has been achieved by a reiterative immersion-rinse approach. Multilayer assemblies consisting of alternate layers of these components have been prepared by the layer-by-layer (LbL) self-assembly technique. Both processes have been carefully monitored by cyclic voltammetry and infrared and UV-visible spectroscopy. Linear increase in the IR and UV-visible light absorbance with the number of deposited layers indicates that well-organized lamellar systems have been elaborated. Size and distribution of Prussian blue nanoparticles in these systems have been investigated by AFM. The effect of the molar concentration of the PB dipping solution on the adsorption process and the distribution of the PB colloids has also been described. Finally, magnetic properties of these assemblies have been studied by low-temperature ESR measurements. Indeed, this new approach of hybrid LbL films opens the way to a new class of nanostructured lamellar compounds. PMID- 16256540 TI - Distribution of water-soluble and surface-active low-molecular-weight species in acrylic latex films. AB - Monodisperse core-shell latices were synthesized, differing in the acrylic acid (AA) content in the particle shell (1 or 4 wt%) and the Tg of the acrylic core (around -40 or 10 degrees C). In a first step, the drying mechanisms of the dialyzed latices were studied by confocal Raman spectroscopy. It was shown that, besides some unexpected features (briefly described in the article), drying occurred in a rather classical way, i.e., simultaneously from top to bottom and from edge to center. Then, the distributions of sulfate ion (SO4) (from sodium sulfate) and sodium dodecyl sulfate (SDS) in the dry latex films were established by confocal Raman spectroscopy and attenuated total reflectance (ATR). The two techniques were complementary. SO4 and SDS distributions were quite different, although presenting some common characteristics. In both cases, repartition of the low-molecular-weight species in the film was even less homogeneous when the AA content was lower and the particle core softer. However, SO4 showed enrichment at the film-substrate interface and depletion at the air side, whereas SDS showed concentration maxima at both interfaces. Interpretations stress the importance of desorption from the particle-water interface, transport by water, size effects, and diffusion. PMID- 16256541 TI - Influence of hydrolyzable metal ions on the interfacial chemistry, particle interactions, and dewatering behavior of kaolinite dispersions. AB - The influence of hydrolyzable metal ions (Mn(II) and Ca(II)) adsorption on the surface chemistry, particle interactions, flocculation, and dewatering behavior of kaolinite dispersions has been investigated at pH 7.5 and 10.5. Metal ion adsorption was strongly cation type- and pH-dependent and significantly influenced the zeta potential, anionic polyacrylamide-acrylate flocculant (PAM) adsorption, shear yield stress, settling rate, and consolidation of kaolinite slurries. The presence of Mn(II) and Ca(II) ions alone led to a systematic reduction in zeta potential due to specific adsorption of positively charged metal ion-based hydrolysis products at the kaolinite-water interface. Metal ion mediated zeta potential changes were reflected by lower dispersion shear yield stresses and improved clarification (higher settling rates) but had no detectable effect on dispersion consolidation. The adsorption of PAM was significantly improved by prior addition of the metal ions. In the presence of Mn(II) or Ca(II) ions, the flocculant adsorption density was enhanced at pH 7.5 for Mn(II) and pH 10.5 for Ca(II). Optimum flocculation conditions, involving partial rather than complete particle surface coverage by both metal ions and flocculant, were identified. As a consequence, the metal ions and flocculant acted synergistically to enhance dewatering, producing particle interactions that were more conducive to high settling rates and greater consolidation of kaolinite dispersions at pH 7.5 than 10.5. PMID- 16256542 TI - Stabilization and characterization of colloidal gas aphron dispersions. AB - Colloidal gas aphrons (CGAs) are finding increasing application in water processing, bioseparation, bubble-entrained floc flotation, separation of oil from sand, etc. This article proposes an effective method of encapsulation to stabilize the CGA bubbles with silicic sol solution for their characterization. The stabilized CGA bubbles can keep shapes and sizes for at least 12 h; even the bubbles smaller than 25 mum can also be stabilized and exist for very long times. Effects of concentration and pH of silicic sol solution on CGA stabilization were studied. The optimal ranges of concentration and pH of silicic sol solution are 0.15-0.25 mol/dm(3) and 7-10, respectively. The bubble distortion behavior in disturbance and size distribution of CGAs were examined by using the stabilization method and photographic techniques. PMID- 16256544 TI - Adhesion in elastic-plastic spherical microcontact. AB - An elastic-plastic adhesion model for a metallic deformable sphere pressed by a rigid flat is presented. Analytical dimensionless expressions for the local separation outside the contact area and for the adhesion force are provided covering a large range of interference values from a point contact to a fully plastic contact. Two main dimensionless parameters of the problem are identified and their effect on the adhesion is investigated. The significance of the adhesion in the elastic-plastic contact force balance is discussed and the regimes where adhesion is important or negligible are pointed out. A comparison of the present results with a previously published approximate elastic-plastic model shows substantial differences in the local separation and in the adhesion force. PMID- 16256543 TI - G1 dendrimers-mediated evolution of silver nanostructures from nanoparticles to solid spheres. AB - With the control of G1 poly(amidoamine) (PAMAM), an evolutionary course of stable colloidal silver from discrete nanoparticles to solid spheres through ultraviolet irradiation reduction of silver nitrate solutions was observed by transmission electron microscopy (TEM). The morphologies of the products depend on the Ag+ concentration. A mechanism of globular assembly was put forward to interpret the evolution of the nanostructures. Powder X-ray diffraction (XRD), electron diffraction (ED) patterns, and X-ray photoelectron spectroscopy (XPS) indicate the presence of cubic symmetry silver. XPS and Fourier transform infrared (FT-IR) spectroscopy confirm that dendrimers have participated in the stabilization and control of Ag nanostructures. In the UV-vis spectra, the intense surface plasmons are centered at 425 and 430 nm corresponding to the shapes of dots and solid spheres, respectively. The solid spheres exhibit excellent catalytic efficiency on the reduction of 2,7-dicholoroflurescein (DCF). PMID- 16256545 TI - Direct measurement of interaction forces between a single bacterium and a flat plate. AB - A technique for precisely measuring the equilibrium and viscous interaction forces between a single bacterium and a flat surface as functions of separation distance is described. A single-beam gradient optical trap was used to micromanipulate the bacterium against a flat surface while evanescent wave light scattering was used to measure separation distances. Calibrating the optical trap far from the surface allowed the trapped bacterium to be used as a force probe. Equilibrium force-distance profiles were determined by measuring the deflection of the cell from the center of the optical trap at various trap positions. Simultaneously, viscous forces were determined by measuring the relaxation time for the fluctuating bacterium. Absolute distances were determined using a best fit approximation to the theoretical prediction for the hindered mobility of a diffusing sphere near a wall. Using this approach, forces in the range from 0.01 to 4 pN were measured at near-nanometer resolution between Staphylococcus aureus and glass that was bare or coated with adsorbed protein. PMID- 16256546 TI - Electrokinetic parameters of colloidal model systems: analysis and comparison between dilute and concentrated dispersions. AB - In the last decades, the interest of many scientists has been focused on the atypical electrokinetic behavior of charged colloidal systems since several studies have shown, in most cases; it is not so ideal as expected. Particularly, two interesting phenomena have not been clearly explained yet. First, the zeta potential magnitude does not decrease monotonically with increasing ionic strength, as expected according to the Gouy-Chapmann model predicts. Second, the zeta potential obtained from different techniques shows discrepancies. More specifically, the zeta potential obtained from streaming potential is lower (in absolute value) than that measured through electrophoretic mobility. However, a recent work has pointed out that these discrepancies seem to disappear if certain conditions (related with the surface charge density) are satisfied. This work also revealed that unexpected results are found when the electric conductivity was used. Spherical polystyrene particles of appropriate particle size and charge density are employed as polymeric colloidal model in the present work. Common and adequate models are used to make clear and easy our theoretical analysis and its interpretation. To test the surface conductance and ionic mobility effects at the solid-liquid interface, both water medium and alcohol-water mixtures are used. PMID- 16256547 TI - Studies on PAN-based carbon fibers irradiated by Ar+ ion beams. AB - In this work, the effects of Ar+ ion beam irradiation on carbon fibers were studied using tensile and surface analytical techniques. The single-fiber pull out test was executed in order to characterize the fiber/epoxy matrix interfacial adhesion. The Ar+ ion beam was irradiated using an ion-assisted reaction (IAR) method in reactive gas conditions under an oxygen environment with 1 x 10(16) ions/cm(2) Ar+ ion dose (ID), 6 sccm blown gas flow rate, and different ion beam energy intensities. From the experimental results, both the interfacial shear strength (IFSS) and fracture toughness (Gi) were found to increase with increasing Ar+ ion irradiation intensity. This was probably due to the fact that Ar+ ion beam irradiation on carbon fibers was effective in altering their surface physical chemistry and structural morphology, resulting in improved interfacial adhesion in the fiber/epoxy matrix. The reliability of single-fiber pull-out test data could be improved by statistical analysis using the Weibull distribution, which served to predict the variation of the mechanical interfacial properties in a composite system. PMID- 16256548 TI - Surface potential and shear plane thickness for a treated concrete particulate. AB - Concrete was treated for the removal of chlorides before the zeta potential (mV), zeta, was measured amphoterically to prepare the adsorbent for onward adsorption of silver on it. The Gouy-Chapman theory, which describes the relationship between the potential at any distance (X) from the surface, Psi(x), and the potential at the surface, Psi(0), was applied to determine the shear plane thickness X(S), and to calculate the surface potential Psi(0). The X(S), for a treated concrete particulate was calculated to be 7.0 A. The Psi(0) calculated was equivalent to the measured zeta potential (zeta), Psi(x) at high electrolyte concentrations, where in general the surface potential greatly deviates from the Nernstian response. PMID- 16256549 TI - Zeta-potential measurement using the Smoluchowski equation and the slope of the current-time relationship in electroosmotic flow. AB - The zeta -potential of a solid-liquid interface is an important surface characterization quantity for applications ranging from the development of biomedical polymers to the design of microfluidic devices. This study presents a novel experimental technique to measure the zeta -potentials of flat surfaces. This method combines the Smoluchowski equation with the measured slope of current time relationship in electroosmotic flow. This method is simple and accurate in comparison with the traditional streaming potential and electrophoresis techniques. Using this method the zeta -potentials of glass and poly(dimethylsiloxane) (PDMS) coated surfaces in KCl and LaCl3 aqueous solutions were measured using several flow channels ranging from 200 to 300 microm in height. The zeta -potential was found to vary from -88 to -66 mV for glass surface and -110 to -68 mV for PDMS surfaces depending on the electrolyte and the ionic concentration. The measured values of the zeta -potential are found to be independent of the channel size and the applied driving voltage and generally are repeatable within +/-6%. PMID- 16256550 TI - Study of molecular interaction in the mixed film of arachidic acid and metal beta diketonate complexes by the "surface ions" method. AB - Molecular interaction is very important for the mechanical properties and application of Langmuir films. In general, fatty acid film is stabilized by certain "subphase ions." In this work, two metal beta-diketonate complexes (M(tmhd)n, tmhd=2,2,6,6-tetramethyl-3,5-heptanedionate) were used as "surface ions" to form stable condensed films with different ratios at the air/water interface. The pi-A isotherms of the mixed films had been measured. The smaller molecular area of the metal beta-diketonate complexes indicated that the metal beta-diketonate complexes form multilayer condensed structures at high pressure at the air/water interface. However, arachidic acid (AA) retained a monolayer structure at high pressure in the mixed system. No considerable phase separations appeared during the compression of the mixed films, which indicated that the mixed films of metal beta-diketonate complexes and AA were miscible and stable. The molecular interaction of the two components in the mixed films was investigated in detail. Mixed systems with the mixing ratio of M(tmhd)n:AA=1:2 were chosen to study the effects of the interaction on the mechanical properties of the mixed films. The molecular interaction between AA and Ce(tmhd)4 is proved to be more significant than that between AA and Sr(tmhd)2, and the pi-A isotherms of the mixed films differ a lot from that of pure AA monolayer. Due to the strong intermolecular interaction, the liquid region disappears in the Ce(tmhd)4/AA mixed films, and dynamic elasticity is improved especially at high surface pressure. On the other hand, the interaction between the AA and the Sr(tmhd)2 is much weaker and the effects of the interaction on the properties (pi-A isotherm and dynamic elasticity) of the mixed films are not so significant, especially at low surface pressure. These results are in accordant with that of the UV spectra analyses. PMID- 16256551 TI - Configuration and photochemical reaction of a bolaamphiphilic diacid with a diazo resin in monolayers and Langmuir-Blodgett films. AB - The monolayer formation of a bolaamphiphile, 1,18-octadecanedicarboxylic acid (ODA), on pure water and the subphase containing a positively charged photoactive 2-nitro-N-methyldiphenylamine-4-diazoniumformaldehyde resin (NDR) have been investigated by pi-A isotherms, pi-t curves, and Brewster angle microscopy (BAM) measurements. It has been revealed that although an unstable monolayer was formed by ODA alone, a stable complex monolayer between ODA and NDR could be formed at the interface through electrostatic adsorption and hydrogen bonding. It has been shown that the ODA formed a U-shaped monolayer at a lower pressure and was converted to a stretched configuration upon compression to a higher surface pressure on the subphase containing NDR. Under UV irradiation at the interface photoreaction can occur in the complex monolayer, which causes shrinkage of the monolayer. Photochemical reactions can also occur in deposited Langmuir-Blodgett films. In reactions occurring at the air/water interface, the two ends of ODA can react with NDR to form an ester containing aromatic rings. This makes the compound more hydrophobic and can easily be stretched without any phase transition upon compression. When the film with U-shaped configuration was deposited onto solid substrates, the configuration could be kept even upon photoirradiation. PMID- 16256552 TI - Distribution kinetics of Ostwald ripening at large volume fraction and with coalescence. AB - Condensation phase transitions from metastable fluids occur by nucleation with accompanying particle growth and eventual Ostwald ripening. During ripening the subcritical particles dissolve spontaneously while larger particles grow and possibly coalesce if their volume fraction is large enough. The classical diffusion-influenced rates are also affected by large particle concentrations and are here described by mass-dependent rates. We represent the kinetics of ripening through growth, dissolution, and biparticle coalescence by a new population dynamics equation for the particle size distribution (PSD). Numerical solutions of the scaled governing equations show that coalescence plays a major role in influencing the PSD when the scaled mass concentration (volume fraction) or number concentration is relatively large. The solution describes the time range from initial conditions to the final narrowing of polydispersity. We show that the time dependence of the average particle mass in the asymptotic period of ripening has a power-law increase dependent on rate expressions for particle growth and coalescence at large values of volume fraction. PMID- 16256553 TI - Rhombohedral-scalenohedral calcite transition produced by adjusting the solution electrical conductivity in the system Ca(OH)2-CO2-H2O. AB - This work is aimed to investigate the effects of the adjustment of the electrical conductivity (kappa25) during the semicontinuous carbonation of Ca(OH)2 suspension (slaked lime) on the morphology of the precipitated calcite (CaCO3) particles. The experiments were carried out at 30, 45, and 60 degrees C. A gradual morphological change from rhombohedral to scalenohedral shapes was produced with an increase of kappa25 from 1 to 7 mS/cm at each temperature. The explanation of this morphological change is given in terms of the increase of both the supersaturation and the ratio between concentrations of charged species containing calcium and carbonate ([Ca]ch/[CO3]ch) in the aqueous phase as the kappa25 set-point increases, prior to the precipitation process. In addition to the rise of the supersaturation this change most probably takes place because the increase of the [Ca]ch/[CO3]ch ratio affects the growth rate of the rhombohedral {104} and scalenohedral {21-1} faces in a different manner: (i) favoring the equality between the surface coverage of Ca2+ and CO3(2-) on the stoichiometric {104} face, thus enhancing the formation of CaCO3(0) growth units and then its growth rate and (ii) inhibiting the growth of the {21-1} face by adsorption of the excess calcium species. PMID- 16256554 TI - 2,3-di-n-decyloxy-6,7-dichloroanthracene (Cl2DDOA), a new low-molecular-mass fluorescent organogelator: physical properties and structures. AB - To extend the family of 2,3-didecyloxyanthracene (DDOA, 1), an organogelator having a rodlike shape, a high polarity, and fluorescing properties, the 6,7 dichloro derivative (Cl2DDOA, 2), was designed and prepared. Compound 2 forms gels in alcohols, nitriles, and alkanes. The electronic absorption spectra of the gel show a finer structure than those of the isotropic solutions, pointing to a specific degree of packing of the molecules; such an aggregation mode is also supported by fluorescence data. The gel-to-sol temperatures (Tm) were determined as a function of gelator concentration and the corresponding enthalpies (DeltaHm) were extracted. Scattering experiments have shown that the molecular packing in aggregates of 2 organogels is less reminiscent of the crystalline state than was the situation with DDOA gels. 2 organogels in butanol are made up of 120 A radius fibers much thinner than those observed in DDOA gels (r ca. 300 A) and with rather monodisperse cross sections. In 1-octanol, dodecane, or cyclohexane, the fibrillar organogel networks involve a broader distribution of the related cross sections through anisometric sections of the fibers and/or formation of bundles. PMID- 16256555 TI - Contribution to the mechanism of liquid membrane oscillators involving cationic surfactant. AB - It is shown that liquid membrane oscillators with cationic surfactants have more complex oscillation patterns than observed previously. The actual details of the oscillations depend strongly on the nature of the membrane material, disclosing even the presence of parallel molecular events. It appears that sampling topology also has a great influence on the observed oscillatory behavior. Variation of oscillation patterns with diffusion path length in the membrane demonstrated the decisive role played by the actual timing of molecular events. The new evidences produced complete usefully the actual views concerning the mechanism of oscillations. PMID- 16256556 TI - Controlling hydrolysis and dispersion of AlN powders in aqueous media. AB - Aqueous suspensions of aluminum nitride (AlN) powders have been prepared in the presence of different surface-active agents, namely, H3PO4 and an anionic surfactant, to avoid the hydrolysis of AlN powders and to enhance dispersion. The most determinant parameters to the hydrolysis process (DeltapH and time of contact) and the stabilization of AlN particles in water (surface crystallinity, surface chemical modification, and surface ionic charge) were seen to be strongly dependent on the acidic agent. The H3PO4 treatment was effective against hydrolysis of AlN due to the formation of a phosphate-based protection layer on the particles' surface, and, although it keeps the pH of the suspension below 4, it does not guarantee a good dispersion. The individual adsorption of the anionic surfactant at the surface of AlN particles suspensions did not completely suppress the hydrolysis but it did enhance the degree of dispersion. A proper combination of the two types of surface-active agents enabled the preparation of AlN aqueous suspensions of relatively low viscosity and high AlN concentration, which can be a good starting point for aqueous-based colloidal shaping techniques or for freeze granulation or spray drying to obtain suitable granulate powder characteristics for dry-pressing technologies. An adsorption mechanism of the surface-active agents onto the particles' surface is proposed and supported by NMR and FT-IR analyses. PMID- 16256557 TI - Thermodynamic fundamental equation of contact lines: selection of independent variables. AB - In this work we review the development of the generalized theory of capillarity. When considering the theory of contact lines, we find that the theory developed by Boruvka and Neumann (BN) requires significant modifications. Their choice of parameters (independent variables) for a line is insufficient for formulating the fundamental equation. Furthermore, there are differential geometric constraints on these geometric parameters but not included in their analysis. As a result, their independent variables are in fact dependent. To describe the geometry of contact lines properly, we present a new set of parameters from the differential geometry viewpoint and subsequently give the fundamental equation for the thermodynamic system of contact lines. PMID- 16256558 TI - A new microcolumn flotation cell for determining the wettability and floatability of minerals. AB - Flotation is one of the most important physicochemical processes for mineral separations and other recovery operations. Flotation machines have been developed since the beginning of the 19th century and are still under intensive research and development. The cell we devised is a combination of the Canadian column flotation cell and the Partridge-Smith cell. The materials used for the construction of the new cell are cheap and use available laboratory accessories and aquarium materials. The cell functions well in terms of its scale, control, and sample requirement. It can be used both in the laboratory for research and in classrooms for demonstrations of experiments. Some of the data obtained by the flotation method using this cell are in good agreement with data measured independently on the same minerals by the contact angles method. The critical values of surface tension of wetting (gamma(c)) for talc, sulfur, and chemically treated surfaces of calcite and barite obtained by the contact angle measurements were 31, 26, 30.5, and 31.2 mN/m, respectively. On the other hand, the gamma(c) values of those minerals, obtained using our new designed flotation cell, were 30, 28, 31.4, and 34.5 mN/m, respectively. The measurements obtained in our experiment are also comparable to those previously published for the same minerals. PMID- 16256559 TI - An analytical solution for determination of small contact angles from sessile drops of arbitrary size. AB - An analytical solution to the capillary equation of Young and Laplace is derived that allows determination of the static contact angle based on the volume of a sessile drop and the wetted area of the substrate. This solution does not require numerical integration to determine the drop profile and accounts for surface deformation due to gravitational effects. Calculation of the static contact angle by this method is remarkably simple and accurate when the contact angle is less than 30 degrees. A natural scaling arises in the solution, which provides indication of when a drop is small enough so as to neglect gravitational influences on the surface shape which, for small contact angles, is generally less than 1 microl. The technique described has the simplicity of the spherical cap approximation but remains accurate for any size of sessile drop. PMID- 16256560 TI - Experimental studies on the geometrical characteristics determining the system behavior of surface tension autooscillations. AB - Autooscillation of the surface tension is a phenomenon related to Marangoni instability periodically arising and fading by dissolution of a surfactant droplet under a water-air interface. A detailed experimental investigation was performed to clear up the influence of the system geometry on development and characteristics of autooscillations. It was found that the aspect ratio is an additional dimensionless parameter that determines the system behavior equally to the Marangoni number. The influence of the cell diameter, capillary immersion depth, and droplet radius on the autooscillation period and amplitude was studied as well. PMID- 16256561 TI - Coalescence during emulsification. 2. Role of small molecule surfactants. AB - An oil-soluble hexadecyl pyrene (HDP) probe is used to monitor coalescence of hexadecane oil-in-water emulsions, during emulsification, in stirred systems and in a high-pressure homogenizer (microfluidizer), when small molecule surfactants are used as emulsifiers. The effect of sodium dodecyl sulfate concentration and salt concentration on the amount of coalescence and final drop size is studied. The behavior of oil-soluble surfactants and mixtures of oil-soluble and water soluble surfactants on emulsification performance is also discussed. For high pressure homogenizers, the drop sizes obtained are found to depend mostly on the ability of surfactants to stabilize the drops against coalescence, rather than their ability to reduce the interfacial tension. Increasing oil phase fractions increase the coalescence rate, because of the increase in collision frequency, which, in turn, impacts the drop size of the homogenized emulsion. PMID- 16256562 TI - NMR diffusion analysis of surfactant-humic substance interactions. AB - Surfactants can be introduced into the environment through wastewater or by direct contamination. Understanding the fate and transport of surfactants in the environment is important in assessing their role as pollutants. Humic substances are complex heterogeneous mixtures of decomposition products of natural organic materials. They are environmentally important because they are known to solubilize and transport organic pollutants. Therefore humic substances are likely to affect the environmental fate of surfactants. Diffusion coefficients measured with pulsed-field gradient nuclear magnetic resonance spectroscopy are used in this study to examine the intermolecular interactions of the surfactants sodium dodecyl sulfate (SDS) and cetyltrimethylammonium bromide (CTAB) in the presence of various humic substances. These results indicate that humic substances enhance the aggregation of SDS prior to micellization with a more pronounced effect observed for the more hydrophobic humic materials. The positively charged surfactant CTAB forms stable ion pairs with the humic substances. PMID- 16256563 TI - Effect of added alpha-lactalbumin protein on the phase behavior of AOT-brine isooctane systems. AB - We have found that the presence of <1 wt% of the globular protein alpha lactalbumin has a significant impact on the equilibrium phase behavior of dilute sodium bis(ethylhexyl) sulfosuccinate (AOT)/brine/isooctane systems. Nuclear magnetic resonance (NMR), Karl Fischer titration, and ultraviolet spectroscopy were used to determine the surfactant, oil, water, and protein content of the organic and aqueous phases as a function of the total surfactant and protein present. As a small amount of alpha-lactalbumin is added to the mixture, there is a substantial increase (up to 80%) in the maximum water solubility in the water in-oil microemulsion phase. Dynamic light scattering measurements indicate that this increase is due to a decrease in the magnitude of the (negative) spontaneous curvature of the surfactant monolayer, as droplets swell in size. As the molar ratio of alpha-lactalbumin to AOT surpasses approximately 1:300, the partitioning of water, protein, and surfactant shifts to the excess aqueous phase, where soluble assemblies with positive curvature are detected by dynamic light scattering. Significant amounts of isooctane are solubilized in these aggregates, consistent with the formation of oil-in-water microemulsion droplets. Circular dichroism studies showed that the tertiary structure of the protein in the microemulsion is disrupted while the secondary structure is increased. In light of these findings, the protein most likely expands to a molten-globule type conformation in the AOT interfacial environment, but does not substantially unfold to become an extended chain. PMID- 16256565 TI - Pressure-driven motion of surfactant-laden drops through cylindrical capillaries: effect of surfactant solubility. AB - The effect of bulk-soluble surfactants on the dynamics of a drop translating through a cylindrical tube under low-Reynolds-number conditions is investigated. Interfacial surfactant adsorption/desorption is modeled according to the Frumkin adsorption framework, and the bulk-insoluble surfactant limit is recovered as the rate of surfactant sorption becomes large compared to that of bulk diffusion. As the equilibrium surface coverage is increased, the mechanism by which drop mobility is reduced changes from uniform retardation at low surface coverage to the formation of a stagnant cap at high surface coverage. For large capillary numbers, the drop does not achieve a steady shape, and eventually it breaks up either through the formation of a penetrating viscous jet of suspending fluid, or by continuous elongation and pinch-off. Surfactants have a destabilizing effect on transient drop shapes by accelerating the formation and development of the penetrating viscous jet that leads to drop breakup. The critical conditions for drop breakup, as well as the mode of breakup, depend on the manner in which the strength of the flow (i.e., the capillary number) is increased. PMID- 16256564 TI - The interaction of an anionic gemini surfactant with conventional anionic surfactants. AB - The interaction in two mixtures of an anionic gemini surfactant having N ,N dialkylamide and carboxylate groups in a molecule, (CH2)2[N(COC11H23)CH(CO2H)CH2(CO2H)]2. 2NaOH (GA), and conventional anionic surfactants have been investigated in 0.1 M NaCl at pH 5.0. The two mixtures are GA/sodium dodecylsulfate (SDS) and GA/sodium N -dodecanoylglutamate (AGS) at a molar fraction of GA, alphaGA = 0.25 . Mixtures of both GA/SDS and GA/AGS exhibit synergism in surface tension reduction effectiveness. The GA/SDS mixture also exhibits synergism in surface tension reduction efficiency and mixed micelle formation, whereas the GA/AGS mixture does not. The interaction in mixed adsorption film formation is stronger than that in mixed micelle formation for the two mixtures. The interaction in the formation of the mixed adsorption film and the mixed micelle for the GA/SDS mixture is stronger in both formations than that for the GA/AGS mixture. The stronger interaction for the GA/SDS mixture may be caused by the combination of the smaller minimum area per molecule at the air/water interface (Amin) of the head groups in the GA molecule and the larger Amin in the SDS molecule. PMID- 16256566 TI - A model for interfacial resistance observed during solubilization with micellar solution. AB - The quantitative behavior of interfacial resistance to mass transfer at an oil water interface in presence of a surfactant, which occurs in addition to the diffusional resistance, is modeled. Successive approximations are used to simplify the detailed behavior to show that the experimentally determined quantitative behavior of solubilization kinetics, is also theoretically justified. The solubility of oil, the frequency of collision of micelles with the oil-water interface, and the resistance to the entry of the solubilizate into the micelle (activation energy) are identified as the key quantities governing this resistance. PMID- 16256567 TI - The diffusion coefficient of a swollen microgel particle. AB - The drag on a permeable particle traversing through a Newtonian liquid is calculated. This is in terms of a single dimensionless group, Da, the Darcy number, which relates the particle permeability to the radius. For small values of the Darcy number the solution reverts to the well-known Stokes drag for smooth hard particles. For larger values of the Darcy number the drag is reduced. This drag allows calculation of the diffusion coefficient of such particles, again as a function of the Darcy number. The results are discussed in relation to microgel particles, which display swelling under certain conditions. The size of such particles is typically measured with dynamic light scattering, which measures the diffusion coefficient of particles and as such the analysis presented here shows the conditions under which dynamic light scattering is accurate. The Darcy number for microgel particles is estimated to be on the order of 10(-7). PMID- 16256568 TI - Architecture and solution properties of amphiphilic polymer brushes with peripheral charged ions. AB - Functionalized poly(ethylene oxide) (PEO) macromonomers (alpha -tertiary amino and omega -methacryloyl groups) were prepared by ring-opening polymerization of live PEO anions end-capped with styrene oxide using 2-[2-(N ,N -dimethylamino) ethoxy]ethanol potassium alkoxides as an initiator with methacryloyl chloride. PEO brushes were synthesized by free-radical homopolymerization of such PEO macromonomers. These brushes were converted into peripherally charged brushes by quaternization. We studied the solution properties of both types of brushes from the viewpoint of charge effect. It was found from dynamic light scattering (DLS) that the polymer brushes formed a single macromolecule in solution due to crowding of side chains. It was speculated from angular dependence measurements that the polymer brushes with large aspect ratios took a geometrical anisotropic conformation such as a cylinder. In methanol with a low dielectric constant, radius of gyration (R(G)), and cross-sectional radius of gyration (R(G,C)) of the polymer brushes with charged side chains were smaller than those of the polymer brushes without charges. In a solvent with a low dielectric constant such as methanol, ionic groups do not dissociate and condense. On the other hand, these physical values in an aqueous solution were somewhat larger than those of the polymer brushes without charges. In water with a high dielectric constant, peripherally charged brushes were strongly stabilized by electric double layers. PMID- 16256569 TI - Adsorption of hydrophobically modified polyelectrolytes at the n-octane/water interface. AB - The interfacial activity of polyelectrolytes carrying alkyl side chains of different length has been studied. Potassium salts of poly(maleic acid-co-1 olefins), PA-n K2 with n=12 , 14, 16, 18, were synthesized, and the interfacial tension at the aqueous solution/n -octane interface was measured as a function of the length of the alkyl side chain. The results show that the interfacial tension lowering, the limiting excess concentration Gamma (m), and the efficiency of adsorption pC (20) depend on the number of methylene groups in the alkyl side chain. According to Rosen the last two parameters define two different contributions to the standard free energy of adsorption: one arises from the distribution of the polymer between the bulk of the solution and the interface Delta G (dist )(0), and another comes from the configuration adopted at the interface Delta G (int )(0). These free energies were plotted as a function of the number of carbon atoms in the alkyl side chain and a linear relation was found for both of them. From these plots contributions of 0.83 and -0.58 per methylene group were determined for Delta G (0)(dist ) and Delta G (0)(int ), respectively. The positive value for the incremental free energy of distribution is attributed to the formation of a polymer micelle which is stabilized by longer alkyl side chains. On the other hand, the negative value for Delta G (0)(int ) indicates that at the interface the polymer adopts a configuration where the hydrocarbon tail is interacting with the octane molecules. PMID- 16256570 TI - Seed-mediated growth approach to shape-controlled synthesis of Cu2O particles. AB - With glucose as reducing agent, Cu2O particles with different size and morphology were synthesized selectively by a seed-mediated growth approach. The major advantage of this process is that Cu(II) cannot be reduced to Cu metal. The size of the Cu2O particles can be controlled by changing the Cu(II) concentration in colloid solution. Square-shaped Cu2O particles (20-700 nm in diameter) were obtained in the presence of PEG 400, and polygonal-shaped Cu2O particles (approximately 1000 nm) were observed upon using triblock copolymer as modifier, while sphere-shaped Cu2O particles with a diameter of 400-700 nm were obtained in the absence of a modifier. PMID- 16256571 TI - Mechanisms of particle removal from silicon wafer surface in wet chemical cleaning process. AB - A quantitative mechanism of particle removal from silicon wafer surfaces by a wet chemical cleaning process is proposed. The particles are removed from the surface due to the combined effects of chemical etching and a net repulsive interaction between the particle and surface. The mechanism suggests that a critical etching depth, which has been determined theoretically, and an optimal etching rate, which can be determined from etching profile calculation, are required for particle removal. The study will help in the optimization of cleaning processes and formulation of superior cleaning solutions. PMID- 16256572 TI - Hydrodynamic instability of a thin viscous film between two drops. AB - Linear stability analysis is employed to derive analytical expressions for the growth rate of disturbances applied to a thin plane-parallel film trapped between two drops. From these expressions, the band of unstable wavenumbers and the "most dangerous" wavenumber are identified for systems in the absence and presence of insoluble surfactant. Marangoni effects are shown to exert a stabilizing influence and reasonably good agreement with experimental observations is found. Subsequent nonlinear analysis indicated amplification of the disturbance growth rate beyond that suggested by linear theory as the film proceeded toward rupture. PMID- 16256573 TI - Investigation of the specific adsorption of sulfate ions on powdered TiO2. AB - The specific adsorption of radiolabeled sulfate ions from perchlorate supporting electrolyte onto TiO2 powder (anatase) has been investigated. The pH and concentration dependence of the adsorption was determined. Langmuir-like adsorption behavior similar to that on other oxides was found. From the pH dependence it follows that the protonation of the surface should precede the specific anion adsorption. The results obtained are compared with those reported for TiO2 bulk electrodes. PMID- 16256574 TI - Electrochemical catalysis with redox polymer and polyion-protein films. AB - Supramolecular redox-active assemblies on electrodes are of fundamental interest and can be used to create functioning devices such as sensors, biosensors, and bioreactors. The ability of redox-active films to mediate electron transfer reactions in 3-D dramatically increases the sensitivity with which target molecules can be determined. Metallopolyion hydrogel films immobilized on electrode surfaces exhibit many properties that are reminiscent of those shown by redox-active proteins. This review discusses the electrochemical properties and applications of such films, including mediating electron transfer between electrodes and oxidase enzymes. In addition, polyion-protein films grown layer by layer have certain advantages in device fabrication, including facilitating direct electron transfer for many proteins, mechanical stability, use of tiny amounts of protein, and control of film architecture. This review presents examples of iron heme proteins in films grown layer by layer by alternate electrostatic adsorption for catalytic reduction of hydrogen peroxide and trichloroacetic acid and for oxidation of styrene. PMID- 16256575 TI - Spontaneous adsorption of viscous and viscoelastic fluids by capillaries and porous substrates. AB - We have developed a new technique to monitor spontaneous adsorption of fluids by porous substrates. The method is based on an optical electronic measuring system providing millisecond resolution. The method capabilities are demonstrated with the example of the absorption of millimeter-size droplets of water and aqueous solutions of polyethylene oxide and polyacrylamide by capillaries. It is shown that polymer additives even in a small amount reduce significantly the rate of adsorption. We have introduced a generalized Lucas-Washburn equation to account for the fluid elasticity. This equation is shown to explain the observed kinetics quantitatively without invoking adjustable parameters. We have derived a modified Bosanquet equation for the initial velocity of penetration, which accounts for the fluid elasticity. This simple formula gives a reasonable estimate of the rate of absorption of small droplets. We report visualization experiments on absorption of water and polymer solutions by sugar cubes as an example of porous substrates. Although the kinetics of droplet adsorption by porous substrates is similar to the kinetics of droplet adsorption by capillaries, the interpretation of experimental data is more complex and requires a plausible hydrodynamic model for lateral spreading in pores. PMID- 16256576 TI - On thermodynamical inconsistency of isotherm equations: Gibbs's thermodynamics. AB - It has been proven that all isotherm equations which include the expression 1 Theta contradict the exact Gibbs thermodynamics. These contradictions have been discussed in detail in the case of the Langmuir (L) equation applied to gas/solid (G/S), solid/liquid (S/L), and gas/liquid (G/L) interfaces. In G/S adsorption the L equation can theoretically be applied only at low equilibrium pressures on condition that vg >> vs . vg is the molar volume of the adsorbed amount in the gas phase and vs is the same in the Gibbs phase. In S/L and G/L adsorption the L equation is practically applicable only in the domain of very low concentrations. The cause of these contradictions (inconsistencies) is that Gibbs thermodynamics takes excess adsorbed amounts into account; however, the L and other isotherm equations calculate with the absolute adsorbed amount. The two amounts may be practically equal to each other when the limiting conditions mentioned above are fulfilled. It is also discussed how these inconsistent isotherm equations can be transformed into consistent ones. PMID- 16256577 TI - Partial specific adsorption of organic molecules in binary mixtures of adsorbents: 2,2'-bipyridine on kaolinite and hematite and 1,10-phenanthroline on montmorillonite and hydroxy-Al montmorillonite. AB - The adsorption of 2,2'-bipyridyl in binary mixtures of sodium kaolinite and hematite at pH 6.6 and of 1,10-phenanthroline in binary mixtures of sodium montmorillonite and hydroxy-Al montmorillonite at pH 7.5 was studied as a function of the equilibrium concentration and composition of the mixture. A model is presented for determining the partial specific adsorption of two adsorbents by each component of the mixture, based on the graphic method for determining partial quantities. The partial specific adsorption values obtained for binary mixtures of Na-kaolinite with hematite and Na-montmorillonite with hydroxy-Al montmorillonite are strongly influenced by the concentration of adsorbates and adsorbents in the mixtures. PMID- 16256578 TI - Coadsorption of Cu(II) and glyphosate at the water-goethite (alpha-FeOOH) interface: molecular structures from FTIR and EXAFS measurements. AB - The coadsorption of Cu(II) and glyphosate (N-(phosphonomethyl)glycine, abbreviated to PMG) at the water-goethite interface was studied by means of batch adsorption experiments, attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy, and extended X-ray absorption fine structure (EXAFS) spectroscopy. The system was investigated over the pH range 3--9 and at total concentrations of 0.9 micromol and 2.2 micromol Cu(II) and PMG per m(2) of goethite. The collective quantitative and spectroscopic results show that Cu(II) and PMG directly interact at the water-goethite interface to form ternary surface complexes. Two predominating complexes have been identified. At pH 4 the IR and CuK-edge EXAFS data indicate a molecular structure where the phosphonate group of PMG bonds monodentately to the surface in an inner sphere mode, while carboxylate and amine groups coordinate to Cu(II) to form a 5-membered chelate ring. Hence, at pH 4, Cu(II) and PMG form a ternary surface complex on goethite with the general structure goethite-PMG-Cu(II). At the highest pH investigated (pH 9), the carboxylate group is still coordinated to Cu(II) but the phosphonate group is present in a relatively free, non-coordinated and/or disordered state. Although the spectroscopic data are not conclusive they indicate the formation of ternary surface complexes with the molecular architecture goethite-Cu(II)-PMG at high pH. PMID- 16256579 TI - The effect of pretreatment of calcite dispersions with anionic sodium polyacrylate on their flocculation behavior induced by cationic starch. AB - The flocculation performance of cationic starches on calcite pretreated with anionic sodium polyacrylate (NaPA) was investigated by measuring the mean particle size and the dynamic mobility of the calcite dispersions. Cationic starches of different molecular weight and degree of substitution were used. By varying the amount of anionic sodium polyacrylate, which has a strong affinity to the calcium carbonate surface, one is able to anionically modify the particles and reverse the charge character of the originally cationic calcium carbonate. By such modification of the charge character of the calcium carbonate dispersion, it is possible to approach the mechanisms of flocculation caused by cationic macromolecules like starch. The importance of different mechanisms of flocculation, such as bridging, charge neutralization, and flocculation induced by polyelectrolyte complexes (PEC), was further investigated in this work. It was found that when the NaPA is completely absorbed at the calcite surfaces the mechanism of the flocculation induced by the starch is mainly bridging flocculation. Excess NaPA in the calcium carbonate dispersion will result in polyelectrolyte complexes formed between the non-absorbed NaPA and the oppositely charged starch polymers. These complexes will in most cases strongly enhance the flocculation due to mainly charge neutralization. Depending on the ratio of non absorbed NaPA and the starch in the aqueous phase, the calcite dispersion is either re-stabilized or more strongly flocculated due to the formed polyelectrolyte complexes. Both the mobility and the particle size measurements support the mechanisms described. It was further demonstrated that the molecular weight and degree of substitution of the starches might be adjusted to control the flocculation behavior. PMID- 16256580 TI - Use of optical tweezers for colloid science. AB - A space-borne optical tweezer apparatus for use with colloidal crystallization experiments has been characterized. The trapping force has been measured as a function of index mismatch between colloidal microspheres and the surrounding fluid and as a function of particle size. This work also presents a method to determine the refractive index of a colloidal microsphere, which is then used to calculate the applied trapping force for the case of an arbitrary background fluid. This is useful for work with dense colloidal suspensions when the usual (e.g., Stokes flow) trap force measurement methods do not apply, as well as microrheological studies of complex soft matter. PMID- 16256581 TI - Cake consolidation in a compression-permeability cell: effect of side-wall friction. AB - A simulation study was made to investigate the transient state stresses, strains, and void ratio distributions in the formation of a filter cake in a compression permeability cell (C-P cell). A finite-element software package, ABAQUS, was used for the simulation and emphasis was placed on the effect of the cake/cell-surface friction. The validity of the simulation was assessed by comparing simulation results with available experimental data. PMID- 16256582 TI - Stability of a dispersion of particles covered by a charge-regulated membrane: effect of the sizes of charged species. AB - The influence of the sizes of charged species on the stability of a colloidal dispersion is investigated theoretically. We consider the case where a particle comprises a rigid core and an amphoteric, charge-regulated membrane layer, which simulates biocolloids and particles covered by artificial membranes. A modified Poisson-Boltzmann equation, which takes the sizes of all the charged species into account, is adopted to describe the electrical field. The effects of other key parameters such as electrolyte concentration, pH, and the valence of counterions on the behavior of a dispersion are also examined. We show that the larger the effective size of the counterions, the greater the stability ratio, which is consistent with experimental observations in the literature. PMID- 16256583 TI - Influence of aluminum doping on titania pigment structural and dispersion properties. AB - The influence of aluminum concentration on the structural properties and rheological behavior of aqueous suspensions of aluminum-doped titania pigment from the chloride process was investigated. The variation in rheological properties correlates with the change in the pigment surface properties, determined from electrophoresis measurements and atomic surface concentrations. Pigment suspensions exhibited a maximum yield stress and viscosity at or near the isoelectric point (iep). The pH of the maximum yield value of the pigment suspension increases with increasing aluminum hydroxyl group density at the particle surface. For pigments with a high aluminum surface concentration, at pH values where the magnitude of the zeta potential was high, a low-viscosity, dispersed suspension was obtained. The pigment with the lowest aluminum concentration, however, retained high yield stresses over a large pH range even when the zeta potential was of considerable magnitude. Pigment particle interactions are chiefly dictated by van der Waals forces and electrostatic repulsive forces, likely to be influenced by heteroaggregation. The aggregate strength would therefore depend upon the proportion and distribution of aluminum and titanium surface groups of the heterogeneous pigment, which will influence both the Hamaker constant and the degree of heteroaggregation. Overall, very small additions to the total aluminum concentration translate to significant aluminum surface concentration disparities and subsequently to large particle interaction differences. PMID- 16256584 TI - Modeling the stability of electroless plating bath--diffusion of nickel colloidal particles from the plating frontier. AB - Electroless nickel (EN) plating is a process in which Ni2+ ions are reduced by hydrogen atoms adsorbed at a fresh Ni surface. However, detaching of a handful of tiny Ni metal particles from a substrate causes the entrance of these particles into the plating solution. The metal particles offer very reactive surfaces for the reduction of Ni2+ ions, which in turn aggravates the detachment, causing a self-accelerated cycle. Eventually the plating solution will be subject to an overwhelming precipitation of Ni black. This paper proposes a one-dimensional diffusion model to explain the dependence of the bath stability on the plating time under different bath loadings. This mathematical model contains Vd, defined as the decomposition volume, a measure to judge chemical stability of a plating solution. To obtain Vd experimentally, a PdCl2 solution was purposely introduced into a model solution (the addition leads to immediate generation of metal particles) until the very moment of onset of massive deposition of colloidal Ni. The Vd data from the experiment were then used to perform simulation in order to complete the model proposed. Other than the effects of bath loadings and plating time, an adsorption model was also created to describe the temperature effect. To coordinate the adsorption model, l-cysteine was used as an adsorbate that plays a deactivation role. The under bump metallization process on patterned silicon wafers has been used to support the main theme of this study. PMID- 16256585 TI - Influence of solvation interactions on the zeta potential of titania powders. AB - The influence of solvation interactions on the surface charge of titania powders was studied by means of linear solvation energy relationships (LSER). The zeta potential (ZP) of titania powders and the effect of water content were determined in a series of alcohols, including methanol, ethanol, 1-propanol, isopropanol, and 1-butanol. The ZPs against the solvents' 13 LSER descriptors were analyzed by principal component analysis (PCA), cluster analysis, multiple linear regression (MLR), and stepwise multiple regression (SMR). Very good correlation between ZP and LSER parameter was obtained. One LSER parameter, relative dielectric constant, epsilon, was used to establish the LSER equation. The ZP of titania powders in pure alcohols can be reasonably calculated by an empirical formula: ZP=13.43-213.63.1/epsilon. The results show that epsilon is the most important factor governing the ZP of titania powders. The results also suggest that the effect of solvation interactions on ZP is a result of the electrostatic interactions between powders and solvents. PMID- 16256586 TI - High-order field electrophoresis theory for a nonuniformly charged sphere. AB - An electrophoresis theory is developed for a rigid sphere in a general nonuniform electric field. The zeta potential distribution and the double-layer thickness are both arbitrary. The zeta potential of the sphere is assumed to be small so that the deformation of the double layer can be neglected. Explicit expressions for the translational and rotational velocities of the sphere are derived in terms of the multipole moments of the zeta potential distribution and the tensor coefficients of the applied electric field. The presence of the kth-order component in the electrical potential field applied to the sphere results in a translation of the sphere only when the sphere possesses the (k-1)th- or (k+1)th order multipole moments of the zeta potential distribution. In addition, the kth order component in the electrical potential field causes a rotation of the sphere only when the sphere possesses the kth-order moment of the zeta potential distribution. As an illustrative example for the utility of our theory, we theoretically devise an electrophoresis analysis scheme for estimating the dipole moment of a dipolar sphere by observing the electrophoretic translation of the sphere in a quadratic potential field. PMID- 16256588 TI - Evaluation of pore size distribution in boundary region of micropore and mesopore using gas adsorption method. AB - This paper discusses an accurate method of pore size distribution evaluation in boundary regions of micropores and mesopores using the gas adsorption process on the basis of the capillary condensation theory, which is liable to be underestimated with the existing BJH and DH methods. A typical nitrogen adsorption isotherm for highly ordered mesoporous silica, which has cylindrical pores with diameter smaller than 4 nm, is considered to be type IV and it is well known for the steep increase of the amount adsorbed through capillary condensation in the region of the relative pressure P/P0 smaller than 0.4. In calculating the distribution of the pore size from the change of the amount adsorbed due to capillary condensation, it is important to accurately predict both the multilayer thickness t of the adsorbed nitrogen molecules and the critical radius rc where capillary condensation occurs. It is necessary to consider the curvature of the adsorption layer-gas phase interface when predicting the multilayer thickness t of nitrogen adsorbed within the pore of highly ordered mesoporous silica. Revision of the Kelvin equation is also required when rc is to be predicted. While the predicted value of t based on the Broekhoff and de Boer theory is matched well with the value of t which is actually measured using highly ordered mesoporous silica, and the predicted value of rc based on the GTKB-Kelvin-cylindrical equation that has been revised considering the effect of the interfacial curvature on the interfacial tension of the adsorption layer-gas phase interface is matched with the value of rc which is actually measured using highly ordered mesoporous silica. A combination method of the Broekhoff and de Boer equation and the GTKB-Kelvin-cylindrical equation is proposed as a means of accurately evaluating, from the nitrogen adsorption isotherm, the pore size distribution in the highly ordered mesoporous silica in boundary region of micropore and mesopore. The proposed new method of pore size evaluation features high accuracy and offers the convenience of obtaining the pore size distribution without repeated calculations by employing the same algorithm as DH method. The pore size predicted by the Halsey equation and the Kelvin equation of the conventional DH method is about 20% smaller than the pore size predicted by the newly proposed evaluation method. PMID- 16256587 TI - A novel dual-impedance-analysis EQCM system--investigation of bovine serum albumin adsorption on gold and platinum electrode surfaces. AB - Both quartz crystal micro-balance (QCM) impedance and electrochemical impedance spectroscopy (EIS) methods are widely used in interface studies. This paper presents details about a new strategy for simultaneous, mutual-interference-free and accurate measurements of QCM impedance and EI, through connecting a suitable capacitance in series with the piezoelectric quartz crystal (PQC) between QCM impedance and EIS measurement instruments. Combined and individual measurements of QCM impedance and EIS during silver deposition gave results comparable with each other, demonstrating the reliability of the proposed method. Bovine serum albumin (BSA) adsorption on gold and platinum electrodes in Britton-Robinson (B R) buffers was investigated, and the Fe(CN)6(3-)/Fe(CN)6(4-) couple was used as an electrochemical probe to characterize BSA adsorption. While the reversibility of Fe(CN)6(3-)/Fe(CN)6(4-) couple on bare Au and Pt electrodes changed very slightly with decreasing solution pH from pH approximately 7 to pH approximately 2, the standard rate constant (ks) of this couple increased abruptly with solution pH below pH approximately 4.5 at a BSA-modified Au electrode, but decreased with solution pH at a BSA-modified Pt electrode. By analyzing the QCM impedance data with a modified BVD equivalent circuit and the EI data with a modified Randle's equivalent circuit, inflexion changes at pH approximately 4.5 were all found at pH-dependent responses of the resonant frequency, the double layer capacitance, the capacitance of the adsorbed BSA layer, the peak-absorbance values of BSA solutions at 277.5 and 224.5 nm, and so on. It was also found that a BSA adsorption layer can effectively inhibit gold corrosion during ferrocyanide oxidation in a ferrocyanide-containing BR solution. Some preliminary explanations of these findings have been given. The proposed method is highly recommended for wider applications in surface science. PMID- 16256589 TI - Determination of coupled solvent mass in quartz crystal microbalance measurements using deuterated solvents. AB - A simple method is described for determining of the contribution of hydrodynamically coupled solvent to the adsorbed film mass determined in a quartz crystal microbalance (QCM) when operated in liquid. The method requires no additional apparatus and utilizes the change in QCM resonant frequency response between measurements made in non-deuterated and deuterated solvents. The mass of coupled water in a polymer film has been determined and is found to agree with that determined by XPS analysis of the dried polymer film. PMID- 16256590 TI - Analysis of tear film rupture: effect of non-Newtonian rheology. AB - We investigate the rupture mechanism of a precorneal thin mucus coating sandwiched between the aqueous tear film and the corneal epithelial surface with a monolayer of surfactant overlying the aqueous layer. The Ostwald constitutive relation is employed to model mucus and a linear equation of state describing the relationship between surface tension and surfactant concentration is adopted. Three nonlinear coupled evolution equations governing the transport of surfactant, mucus, and total liquid layer thicknesses, based on lubrication theory and a perturbation expansion technique, have been derived. The resulting equations are solved numerically in order to explore the influence of the rheological properties of mucus, aqueous-mucus thickness ratio, aqueous-mucus interfacial tension, Marangoni number, and surfactant concentration on both the onset of instability and tear film evolution in the presence of van der Waals interactions, which could rupture the tear film. Our results reveal that the influence of rheological properties, aqueous-mucus thickness ratio, and interfacial tension on the time required for film rupture can be significant and varies considerably, depending on the magnitude of the Hamaker constants governing the strength of the van der Waals forces. PMID- 16256591 TI - Numerical simulation and experimental verification of particle coagulation dynamics for a pulsed input. AB - Mathematical simulation of particle coagulation dynamics was carried out using improved sectional modeling techniques for a system with a pulsed input of primary particles. The methodological improvement included the modification of the size density function based on a realistic assumption of particle size distributions, the application of a new and comprehensive curvilinear collision model, and special adjustment for the mass transfer of a doublet of particles that were very different in size. The simulation results demonstrated that the rectilinear model over-predicted the rate of particle coagulation and that the degree of over-prediction increased as the particles increased in size and the system became more heterogeneous. The coagulation rate increased remarkably as the fractal dimension of the particle aggregates decreased. The curvilinear model and the fractal scaling relationship in place of the rectilinear model and the Euclidean sizing geometry are two important modifications to the conventional Smoluchowski modeling approach. However, both modifications, rather than only one of them, should be applied together to produce more accurate and realistic simulations of coagulation dynamics. As indicated by the simulation, the importance of fluid shear rate to particle coagulation is reduced according to the curvilinear model compared to that previously described with the rectilinear model. As particles increased in size, the role of shear rate in coagulation became even less significant according to the curvilinear view of particle collisions. The results of numerical simulations in terms of the evolution of particle size distributions compared reasonably well with the observations of the jar-test coagulation experiments, which suggested the applicability of the modeling system, including the modified curvilinear-fractal approach, established in the present study. PMID- 16256592 TI - Gapped gapless packing structures. AB - The assumption of a gapless packing structure has previously been used to obtain the density and partial coordination numbers of a random mixture of hard spheres in the maximally dense regime. Here we extend the notion of a gapless packing structure to allow the determination of the characteristics of a packing away from maximal density by adding an appropriate number of void spherical elements. A gapless packing is then considered in which the void and solid spherical elements are assumed to be indistinguishable except for the purposes of calculating packing fraction and coordination number. We utilize the notion of specific volume to generate a one-parameter family of void distributions to obtain a set of coupled integral equations, which are solved numerically. Monodisperse and bi-disperse packings are investigated for packing fractions ranging from rho=0.26 to 0.78. Results are shown to be comparable to experiments and the effect of varying packing fraction on coordination numbers is shown to be invariant with respect to number distribution. A linear relationship between coordination number and packing fraction is elucidated for moderate to low packing fractions. Maximum and minimum random packing fractions are also discussed. PMID- 16256593 TI - On the evaluation of the surface free energy of porous and powdered solids from imbibition experiments: equivalence between height-time and weight-time techniques. AB - A study of the equivalence between the height-time and weight-time experimental techniques, based on the imbibition of liquids into porous or powdered solids, has been carried out, determining under what conditions the deductions about the surface free energy of these solids made from the analysis of the experimental results obtained as weight versus time are similar to those deduced from the analysis of height versus time. Concretely, we have proved that only if the particular values of the porosity of the solid determined from the proper technique of imbibition, named the effective porosity, are taken into account, the analysis of both type of experimental measurements leads to the same conclusions about the surface free energy of the porous and powdered solids. In order to exemplify this, capillary rise measurements have been carried out by means of these two techniques with different liquids on silica gel layers. PMID- 16256594 TI - Porous properties of activated carbons from waste newspaper prepared by chemical and physical activation. AB - Activated carbons were prepared from old newspaper and paper prepared from simulated paper sludge by chemical activation using various alkali carbonates and hydroxides as activating reagents and also by physical activation using steam. In the chemical activation, the influence of oxidation, carbonization, and activation on the porous properties of the resulting activated carbons was investigated. The specific surface areas (S(BET)) of the activated carbons prepared by single-step activation (direct activation without oxidation and carbonization) were higher than those resulting from two-step activation (oxidation-activation and carbonization-activation) and three-step activation (oxidation-carbonization-activation) methods. The S(BET) values were strongly dependent on the activating reagents and the activating conditions, being >1000 m(2)/g using K(2)CO(3), Rb(2)CO(3), Cs(2)CO(3), and KOH as activating reagents but <1000 m(2)/g using Li(2)CO(3), Na(2)CO(3), and NaOH. These differences in S(BET) values are suggested to be related to the ionic radii of the alkalis used as activating reagents. The microstructures of the higher S(BET) samples show a complete loss of fiber shape but those of the lower S(BET) samples maintain the shape. In the physical activation, the porous properties of the activated carbons prepared by the single-step method were examined as a function of the production conditions such as activation temperature, activation time, steam concentration, and flow rate of the carrier gas. The maximum S(BET) and total pore volume (V(P)) were 1086 m(2)/g and 1.01 ml/g, obtained by activation at 850 degrees C for 2 h, flowing 20 mol% of steam in nitrogen gas at 0.5 l/min. A correlation was found between S(BET) and the yield of the product, the maximum S(BET) value corresponding to a product yield of about 10%. This result is suggested to result from competition between pore formation and surface erosion. Compared with chemically activated carbons using K(2)CO(3), the porous properties of the physically activated carbons have lower S(BET) and V(P) values because of the smaller size and lower volume of their micropores. On the other hand, they retain the original fiber shape and the paper sheet morphology after activation. PMID- 16256595 TI - Adsorption properties of activated carbon from waste newspaper prepared by chemical and physical activation. AB - Adsorption properties of activated carbons prepared from waste newspaper by chemical and physical activation were investigated using water vapor, ammonia, methane, and methylene blue (MB) as adsorbents. The water vapor adsorption isotherms show type V behavior and the maximum vapor adsorption of the chemically and physically activated products is about 1050 and 450 ml/g, respectively. The higher water vapor adsorption of the chemically activated products is attributed to the higher specific surface area (S(BET)) and greater hydrophilic activity (arising from the surface oxygen-containing functional groups) than in the physically activated products. The adsorption of ammonia and methane was measured by temperature-programmed desorption (TPD). NH(3) adsorption is found to be higher in the chemically activated product than in the physically activated product while methane adsorption is slightly higher in the physically activated products even though these have lower S(BET) values. In the MB adsorption, the chemically activated products show higher adsorption (390 mg/g) than the physically activated product. These results are suggested to be related to the surface characteristics. PMID- 16256596 TI - Evaluation of the "DSPM" model on a titania membrane: measurements of charged and uncharged solute retention, electrokinetic charge, pore size, and water permeability. AB - The DSPM (Donnan steric partitioning pore model) was evaluated in the case of a titania membrane with "nanofiltration properties" by measuring the electrokinetic charge, pore size, and water permeability of the membrane, along with charged and uncharged solute retention. The zeta potential values (zeta) were determined from measurements of the electrophoretic mobility (EM) of titania powder forming the filtering layer of the membrane. Zeta potential values were converted into membrane volume charge (X) by assuming two limiting cases: a constant surface charge (sigma(s)(cst)) and a constant surface potential (psi(s)(cst)). The mean pore radius and thickness/porosity ratio of the membrane were determined by permporometry and from water permeability measurements, respectively. Retention measurements were carried out as a function of the permeate volume flux for both neutral solutes (polyethylene glycol PEG of different size) and salts (KCl, MgSO4, K2SO4, and MgCl2) at various pH values. Ionic retentions showed minimum values near the IEP of the membrane. Retention data were analyzed using the DSPM. Very good agreement was found between the pore radius calculated by the model and that determined by permporometry. X values calculated from fitting retention data using the DSPM were also in satisfactorily agreement with X values calculated from EM measurements assuming a constant surface potential for a large pH range. Furthermore, the DSPM leads to X values (X(DSPM)) between those calculated from EM (X(EM)) using the two limiting bounds. In other words, X(DSPM) was higher than X(EM) assuming psi(s)(cst) at pH values far from the isoelectric point (IEP) and lower than X(EM) assuming sigma(s)(cst). These results show that the DSPM is in qualitative agreement with the charge regulation theory (increase of the pore surface potential and decrease of the pore surface charge density with decreasing the pore size). On the other hand, the thickness/porosity ratio of the membrane calculated from solute retention data differed significantly from that determined from water permeability measurements. Moreover, a single value of Deltax/Ak could not be determined from PEG and salt retention data. This means that the Deltax/Ak parameter loses its physical meaning and includes physical phenomena which are not taken into account by the DSPM. Nevertheless, the model satisfactorily predicted the limiting retention, as this is not influenced by the Deltax/Ak parameter. PMID- 16256597 TI - On the stability of double bubbles and double drops. AB - The stability problems for equilibrium standard "double bubble" and "double drop" configurations are considered. Recent work has shown that a double bubble is stable to volume-preserving perturbations. In this paper, the stability to perturbations that do not conserve the volumes of the individual bubbles is examined. It is shown that a double bubble shape is also stable to these perturbations and, thus, is stable to arbitrary perturbations. The analysis is based on the principle of minimum total energy. A variational principle is used to formulate the stability problem for an equilibrium double drop configuration formed under zero gravity by two drops of immiscible incompressible liquids. PMID- 16256598 TI - Fluorescence depolarization and contact angle investigation of dynamic and static interfacial tension of liquid crystal display materials. AB - Interfacial interactions control two processes empirically known to be critical for molecular anchoring in twisted nematic liquid crystal displays technology (TN LCDs): surface treatment and filling procedure. Static and dynamical interfacial tensions (Gamma(SL)) between liquids and several substrates with similar roughness were observed respectively by contact angle (theta(c)) of sessile drops and by fluorescence depolarization of thin liquid films flowing at high velocity. Gamma(SL) decreased when glass was coated with tin dioxide and increased with polyvinyl alcohol (PVA) deposition. Drops were circular for all substrates except rubbed PVA, where they flowed spontaneously along the rubbing direction, reaching an oblong form that had theta(c) parallel and perpendicular to the rubbing direction respectively greater and smaller than theta(c) for non-rubbed PVA. This is attributed to polar group alignment generating an asymmetric Gamma(SL) distribution with nanometric preferential direction, inducing a capillary-like flow. Polarization and anisotropy maps for high-velocity flow parallel to the PVA rubbing direction showed an increase in the net alignment of molecular domains and a widening of the region where it occurred. This is attributed to preferential anchoring in the downstream direction, instead of in several directions, as for non-rubbed PVA. This explains why filling direction is crucial for TN-LCDs homogeneous behavior. PMID- 16256599 TI - Effect of equimolar salt to decyltrimethylammonium decyl sulfate on vesicle formation and surface adsorption. AB - The aqueous solution of mixture of sodium decyl sulfate (SDeS) and decyltrimethylammonium bromide (DeTAB) has been found to form equilibrium multilamellar vesicles (MLV) spontaneously. We measured the surface tension of the aqueous solution of 1:1 mixture of SDeS and DeTAB as a function of temperature T at various molalities m under atmospheric pressure. The surface density, the entropy of adsorption and the entropy of vesicle formation are evaluated and compared with those of the decyltrimethylammonium decyl sulfate (DeTADeS) aqueous solution system to investigate the role of small counterions in the mechanism of equilibrium vesicle formation. The saturated surface density Gamma (H,C ) vs T curve of the SDeS/DeTAB system sits below that of the DeTADeS system. Therefore, sodium and bromide ions are negatively adsorbed and nevertheless, they neutralize the electric charge of the decyl sulfate ion DeS(-) and the decyltrimethylammonium ion DeTA(+) to some extent to weaken the electrostatic attraction between the polar head groups in the adsorbed film. The net surfactant concentration required for vesicle formation was larger in the SDeS/DeTAB system. Hence, the electrostatic attraction between the polar head groups of the surfactant ions which is one of the major driving forces for vesicle formation is weakened by the presence of the counterions Na(+) and Br(-). Small but distinct changes in the surface density and the entropies of MLV formation of the SDeS/DeTAB system from those of the DeTADeS system were also found. PMID- 16256600 TI - The structure of mixed nonionic surfactant monolayers at the air-water interface: the effects of different alkyl chain lengths. AB - The structure of mixed nonionic surfactant monolayers of monodecyl hexaethylene glycol (C10E6) and monotetradecyl hexaethylene glycol (C14E6) adsorbed at the air water interface has been determined by specular neutron reflectivity. Using partial isotopic labeling (deuterium for hydrogen) of the alkyl and ethylene oxide chains of each surfactant, the distribution and relative positions of the chains at the interface have been obtained. The packing of the two different alkyl chain lengths results in structural changes compared to the pure surfactant monolayers. This results in changes in the relative positions of the alkyl chains and of the ethylene oxide chains at the interface. The role of the alkyl chain length is contrasted with that of the ethylene oxide chain length, determined from results reported previously on the nonionic surfactant mixture of monododecyl triethylene glycol (C12E3) and monododecyl octaethylene glycol (C12E8). PMID- 16256601 TI - Transfer rate of several tris(beta-diketonato)iron(III) complexes at the bulk aqueous--Triton X-100 micellar interface: effects of substituent groups in the extractants. AB - The transfer rate of tris(beta-diketonato)iron(III) complexes with acetylacetone, benzoylacetone, trifluoroacetylacetone, benzoyltrifluoroacetone, and 2 thenoyltrifluoroacetone at the water-Triton X-100 micellar interface was measured by stopped-flow spectrophotometry. The forward transfer rate was first-order with respect to the micellar concentration; the rate-determining step was concluded to be the capturing process of the collided complex at the micellar surface. The value of the rate constant varied with the substituent groups in the extractants; several effects of the substituent groups on the transfer rate were observed. The "mantlephobicity" of the trifluoromethyl group because of its low wettability, the hydrophobicity of the phenyl and thienyl groups in light of their bulkiness, the dilution of the negative efficiency of the trifluoromethyl group at the capturing by enlarging the complex surface with these bulky groups, and the "mantlephilicity" of the thienyl group from its interaction with the hydrogen bonded network of the micellar "mantle" were estimated. In contrast, the backward rate was independent of the micellar concentration, and the order could be explained by the distribution of the tris-complexes between the "core" and "mantle" of the micelle. The "core-mantle" double-layer structure of the micelle would be an important key for the behavior at the interface. PMID- 16256602 TI - Modeling of spontaneous penetration of viscoelastic fluids and biofluids into capillaries. AB - A theoretical model was developed to describe the dynamics of spontaneous penetration of viscoelastic fluids into capillaries. The model agrees quantitatively with recent experiments on absorption of droplets of polymer solutions by glass capillaries [A.V. Bazilevsky, K.G. Kornev, A.N. Rozhkov, A.V. Neimark, J. Colloid Interface Sci. (2003)]. The rate of penetration progressively reduces with the increase in fluid elasticity. Analysis revealed two main contributions to the viscoelastic drag of the liquid column: (i) viscous resistance, which is independent of fluid elasticity, and (ii) viscoelastic resistance, known as the Weissenberg effect. We analytically derived an augmented Bosanquet equation for the maximal velocity of penetration by balancing capillary, inertia, and viscoelastic forces. For slow creep of a liquid column, the Lucas-Washburn equation was modified by accounting for the Weissenberg effect. A series of numerical calculations were performed to demonstrate characteristic features of absorption of fluids at different conditions. This article also discusses some problems specific to absorption of biofluids. We show that deformations of cell membranes in the external converging flow may cause their rupture at the pore entrance. PMID- 16256603 TI - Rheological properties and particle behaviors of a non-dilute colloidal dispersion composed of ferromagnetic spherocylinder particles subjected to a simple shear flow (analysis by means of mean-field approximation). AB - We have investigated the rheological properties and the orientational distributions of particles of a non-dilute colloidal dispersion, which is composed of ferromagnetic spherocylinder particles, subject to a simple shear flow. The mean-field approximation is applied to take into account the interactions between spherocylinder particles. The basic equation of the orientational distribution function has been derived from the balance of the torques (including the term due to the mean field approximation) acting on the particle in an applied magnetic field; this is an integrodifferential equation. Then, the governing equation has been solved by means of the method of successive approximation and Galerkin's method. The results obtained here are summarized as follows. For the case of strong magnetic interactions between particles, the particle exhibits a sharp peak of the orientational distribution even for a weak applied magnetic field. In this case, the mean magnetic moment of the particle becomes large, which leads to strong interactions between the applied magnetic field and the particle. Thus, the particle tends to point to the magnetic filed direction under these situations. Also, in this case, a large increase in viscosity is obtained due to such a restriction concerning the particle orientation. PMID- 16256604 TI - PAA/PEO comb polymer effects on rheological properties and inter-particle forces in aqueous silica suspensions. AB - The effects of a poly(acrylic acid) (PAA)-poly(ethylene) (PEO) comb polymer dispersant on the rheological properties and inter-particle forces in aqueous silica suspensions have been studied under varying pH conditions. The comb polymer was found to adsorb more strongly under acidic than basic conditions, indicating that the PAA backbone of the copolymer preferentially adsorbs onto silica surfaces with the PEO "teeth" extending out from the surface into the solution. In the presence of low concentrations of copolymer, the silica suspensions were stable due to electrostatic repulsions between the silica surfaces. At higher copolymer concentrations and under neutral and basic conditions, where the copolymer interacted only weakly with silica, the suspensions showed a transition from a dispersed to weakly flocculated state and attractive forces were measured between silica surfaces. Under acidic conditions, the silica dispersion also destabilized at intermediate copolymer adsorbed density and then was re-stabilized at higher adsorbed coverage. The silica suspensions were stable at high copolymer coverage due to steric repulsions between the particles. The destabilization at intermediate coverage is thought to be due to polymer bridging between particles or possibly depletion forces. PMID- 16256605 TI - Isomerization of cationic azobenzene derivatives in dispersions and films of layered silicates. AB - Photoisomerization reactions of cationic azobenzene dyes in solutions, dispersions, and films of layered silicates were studied by visible (Vis) spectroscopy. The dyes isomerized reversibly from thermodynamically more stable trans-isomers to cis-isomers when irradiated with ultraviolet (UV) light. Observed trends were compared with the optical changes of the dyes that occurred as a consequence of their adsorption at the silicate surface. Small fractions of the dyes are likely to have isomerized during the adsorption process, even without the UV-light irradiation. The aggregation of the dyes was another reaction taking place at the surface of the silicates. The extent of the UV-light induced isomerization reactions was reduced for the adsorbed dye cations. The reaction proceeded readily for a dye with monovalent cations. However, the photoisomerization was practically negligible in both dispersions and films of layered silicates for a dye with bivalent cations, whereas the isomerization proceeded in solution. This phenomenon was interpreted in terms of the attractive electrostatic forces between the substrate and the dye cations, which hindered the isomerization reaction. The layer charge of silicates affected the orientation of the dye cations as observed by X-ray diffraction (XRD) measurements. However, the choice of silicate did not significantly affect the fundamental aspects and the described basic trends of the UV-light-induced isomerization reaction. PMID- 16256606 TI - Decarboxylation of 6-nitrobenzisoxazole-3-carboxylate in aqueous cationic micelles: kinetic evidence of microinterface property changes. AB - We studied decarboxylation of 6-nitrobenzisoxazole-3-carboxylate, 1, as a kinetic probe to investigate microinterface properties of aqueous micelles formed by cationic surfactants of increasing head group bulk, i.e., cetyltrialkylammonium bromide, with alkyl=Me (CTABr), Et (CTEABr), n-Pr (CTPABr), n-Bu (CTBABr) and p octyloxybenzyltrialkylammonium bromide surfactants with alkyl=Me (pOOTABr), n-Pr (pOOTPABr), and n-Bu (pOOTBABr), and the longer p dodecyloxybenzyltrimethylammonium bromide (pDoTABr) at concentrations higher than 0.05 M. The pseudophase kinetic treatment fails to fit the data that show anomalies with abrupt increases in k(obs) for CTPABr and CTBABr (but not for CTEABr) and with smooth and continuos increase of k(obs) for all p alkyloxybenzyltrilakylammonium bromides. Abrupt and successive modifications of the micellar interface properties, undergoing only when the polar head or the alkyl chain have some covalent structure, account for the observed kinetic behavior. PMID- 16256607 TI - Numerical calculation of the electrophoretic mobility of a spherical particle in a salt-free medium. AB - By extending an approximate theory of the electrophoretic mobility of dilute spherical colloidal particles in a salt-free medium containing only counterions (H. Ohshima, J. Colloid Interface Sci. 248 (2002) 499--503), a systematic numerical method is given for the calculation of the electrophoretic mobility, which is based on an iteration method. We assume that each sphere is surrounded by a spherical free volume, within which counterions are distributed so that electro-neutrality is satisfied. The electrophoretic mobility is found to be determined mainly by the pressure due to the counterions at the outer surface of the free volume. It is shown how the mobility values deviate from those expected from Huckel's formula for high particle charges or zeta potentials because of the counterion condensation effect. PMID- 16256608 TI - Under pressure: predicting pressurized metered dose inhaler interactions using the atomic force microscope. AB - Drug particulate interactions in pressurized metered dose inhalers (pMDI) may lead to a decrease in aerosolization efficiency and subsequent efficacy in patient treatment. The interactions between salbutamol sulfate (commonly used in Ventolin pMDIs) and a series of pMDI canister materials were investigated using the atomic force microscope (AFM) colloid probe technique. Approximately 4000 individual force-distance curves were determined for a drug probe and three surfaces (10 x 10 mum areas) in situ, in a model propellant. The area under each force-distance curve was integrated to obtain separation energy values. Median separation energy values followed the rank order borosilicate glass > aluminum > PTFE, suggesting PTFE to be the most suitable canister coating. PMID- 16256609 TI - The dynamic nature of contact angles as measured by atomic force microscopy. AB - Atomic force microscopy appears to be a useful tool for determining the contact angle for small particles. It is shown in this paper that the contact angle of a spherical polyethylene particle changes with the speed of the AFM piezoelectric translator. Such dynamic behavior of the contact angle and other uncertainties such as the position of the three-phase contact on the particle surface during bubble-particle interaction make it difficult to decide whether or not the AFM single-particle contact angle can be used to describe the hydrophobic state of the particle surface. PMID- 16256610 TI - Comment on "Adsorption of fluoride, phosphate, and arsenate ions on a new type of ion exchange fiber," by R.X. Liu, J.L. Guo, and H.X. Tang. PMID- 16256611 TI - Surface heterogeneity of passively oxidized silicon carbide particles: vapor adsorption isotherms. AB - The surfaces of silicon carbide particles subjected to two different passive oxidation treatments have been characterized by immersion calorimetry and vapor adsorption techniques. Surface enthalpies and surface free energies have been computed using semiempirical models and are compared to theoretical estimations. The surface entropy term appears higher than in the case of other solids studied with the same analysis. The definition of the surface entropy term is discussed in order to explain the discrepancy between calculation and experiment. An explanation of results is proposed, which is related to the constitution of silicon oxide layers at the surface of silicon carbide, a fact demonstrated by previous XPS measurements. PMID- 16256612 TI - Equilibrium study of selected divalent d-electron metals adsorption on A-type zeolite. AB - The objective of the presented study was to investigate the adsorption of Cu, Co, Mn, Zn, Cd and Mn on A-type zeolite. The isotherms for adsorption of metals from their nitrates were registered. The following adsorption constants K of metals were found: 162,890, 124,260, 69,025, 16,035, 10,254, and 151 [M(-1)] for Cu, Co, Mn, Zn, Cd, and Ni, respectively, for the concentration range 10(-4)-10(-3) M. On the other hand, the investigation of pH influence on the distribution constants of metals showed that the adsorption of metals proceeds essentially through an ion-exchange process, surface hydrolysis, and surface complexation. The supplementary results from DRIFT, scanning electron microscopy, and X-ray diffraction methods confirmed the presumption about the possible connection between the electronic structure of divalent ions and their adsorption behavior, showing that ions with d5 and d10 configurations such as Mn2+, Zn2+, Cd2+, with much weaker hydrolytic properties than Cu2+ and Ni2+, strongly interact with the zeolite framework and therefore their affinity to the zeolite phase is much stronger when compared with that of the Ni2+ ion, but at the same time not as strong as the affinity of the Cu2+ ion, the latter forming a new phase during the interaction with zeolite framework. For Zn2+, during inspection of the correlation between the proton concentration H/Al and zinc concentration Zn/Al on the zeolite surface, the formation of the surface complex [triple bond]S-OZn(OH) was proposed. A correlation between the heterogeneity of proton concentrations H/Al on Me-zeolite surfaces and the hydrolysis constants pKh of Me2+ ions was found. PMID- 16256613 TI - Microcolumn test and model analysis of activated carbon adsorption of dissolved organic matter after precoagulation: effects of pH and pore size distribution. AB - Microcolumn adsorption experiments were conducted to generate breakthrough profiles of dissolved organic matter (DOM) remaining after coagulation treatment of a naturally colored surface water for three coal-based activated carbons (ACs) and four water pH levels. A plug-flow homogeneous surface diffusion model was applied to determine the intraparticle surface diffusivities of the DOM at different AC-pH combinations. It was found that, for all three ACs, the removal of DOM increased as pH decreased and the increasing extent changed with the ACs used. The pH dependency of the column performance seemed to be attributed more apparently to pH's capability in changing the zeta potential of AC particles. In addition, at all pH levels, the column performance varied markedly with the ACs used. Correlation analyses of the accumulated amounts of DOM onto all three ACs with corresponding pore volumes in several divided pore size regions clearly indicated that pores with sizes 30-100 A were more effective in adsorbing organic macromolecules. Furthermore, based on model simulations, the sensitivity of bed performance to equilibrium and kinetic parameters was quantified by conducting variance analyses with a four-way classification method. PMID- 16256614 TI - A new insight into the adsorption of bovine serum albumin onto porous polyethylene membrane by zeta potential measurements, FTIR analyses, and AFM observations. AB - The mode of adsorption of bovine serum albumin (BSA) on porous polyethylene (PE) membrane was studied as a function of time and concentration, which may contribute to the surface coverage. An improved physical model for adsorption is initiated based on the results of the adsorptional and desorptional measurements, FTIR analysis, and AFM observations as well as streaming potential measurements. The results obtained indicate that the adsorptional mode depend on both time and concentration. It is shown that a critical concentration (about 1000 ppm here) exists in the adsorptional process. Below this concentration, the adsorption seems to be conducted in a normal side-on way but time elapse gives rise to greater conformational change than concentration increase; above this concentration, the aggregation of protein molecular plays a decisive role and the adsorption is in an aggregation way, which is similar to end-on, but a relative large gap between the adsorbed molecules exists due to aggregation. This conclusion is general and can be expected to apply in other globular protein hydrophobic porous surface systems. PMID- 16256615 TI - Study of the interaction between europium (III) and Bacillus subtilis: fixation sites, biosorption modeling and reversibility. AB - In order to elucidate the underlying mechanisms involved in the biosorption of metal ions, potentiometric titrations, complexation studies, and time-resolved laser-induced fluorescence spectroscopy (TRLFS) measurements were used to characterize the interaction between Eu(III) and Bacillus subtilis. The reversibility of the interaction between Eu(III) and Bacillus subtilis was studied by a cation-exchange technique using the Chelex resin. For complexation studies in the presence of 0.15 mol/l of NaCl, the metal ion, the biomass, concentrations and the pH were varied. The adsorption data were quantified by a surface complexation model without electrostatic term. The data on the Eu(III)/B.subtilis system at pH 5 were satisfactorily described by one site at which Eu(III) was bound through one carboxylic function of the bacteria. With increasing pH, another site should be considered, involving a phosphate-bound environment. This was partially confirmed by time-resolved laser-induced fluorescence spectroscopy. In addition to this, it was evidenced that the site availability was dependent on the nature of the cation, i.e., a proton or Eu(III). Finally, it was shown that, at pH 5, the Eu(III)/Bacillus subtilis equilibrium was reversible. PMID- 16256616 TI - Adsorption of a cationic gemini surfactant from aqueous solution onto aluminosilicate powders of the MCM-41 type: effect of pore size and co-adsorption of phenol. AB - The adsorption of cationic gemini dodecanediyl-alpha,omega bis(dodecyldimethylammonium) bromide (i.e., C12C12C12) from aqueous solution onto aluminosilicate powders of the MCM-41 type (referred to as SiAl32dx, where x is the mean pore diameter in A) has been studied at 298 K under the conditions of free pH of the aqueous phase. Macroporous silica gel XO15M was used for comparative purposes. The discussion was based on the interpretation of experimental adsorption isotherms and differential enthalpies of displacement obtained on various solid samples. For the hydrogen-exchanged SiAl32d28 sample (i.e., H+-SiAl32d28), the adsorption of bromide counterions Br- at the solid solution interface, the isotherm of the pH evolution in the equilibrated supernatant liquid, and the electrophoretic mobility of the solid particles coated with the adsorbed C12C12C12 were additionally measured. The uptake of phenol (PhOH) by a surfactant-solid system from a 1.5 mmolkg(-1) PhOH solution and the related thermal effect of displacement onto SiAl32d50 were quantified with the use of the solution depletion technique combined with UV spectrophotometry and the titration calorimetry technique. Titration calorimetry was also used to determine the molar enthalpy change accompanying micellization of C12C12C12 in pure deionized water and in a 1.5 mmolkg(-1) PhOH solution at 298 K. The adsorption of C12C12C12 occurs simultaneously on the external surface and on the pore walls and it is a strongly co-operative phenomenon. Surfactant aggregates forming at adsorption saturation are thought to be composed of the adsorbed surfactant units having their cationic head groups mostly oriented outward with respect to the solid surface. Therefore, they can provide co adsorption sites for polarisable phenol molecules. On average, there is one phenol molecule retained for one gemini cation adsorbed. Transfer of phenol from the aqueous phase to either the bulk micelles or to the interfacial aggregates is enthalpically favourable. PMID- 16256617 TI - Effect of depletion interactions on transport of colloidal particles in porous media. AB - The influence of depletion interactions on the transport of micrometer-sized, negatively charged polystyrene latex particles through porous media was studied by analysis of particle breakthrough curves as a response to short-pulse particle injections to the inlet of a packed column of glass beads. The column outlet latex particle concentration profiles and the total amount of particles exiting the column were determined as a function of the concentration of small, silica nanoparticles in the solution and the bulk flow rate. Because of similar charges, the silica particles do not adsorb to either the latex particles or glass beads and thus induce an attractive depletion force between the latex particles and glass bead collectors. The total column outlet latex particle amount was calculated by integrating the measured breakthrough concentration curve and compared to the known amount of injected particles at the column inlet. It was found that the particle recovery was a decreasing function of the silica nanoparticle concentration and the carrier fluid residence time, and an increasing function of the velocity in the bed. In addition, removing the silica nanoparticles from the flowing solution caused a second outlet peak to appear, suggesting that some of the polystyrene particles were captured in secondary energy wells. The experimental data were interpreted using the predicted potential energy profile between a single particle and a glass bead, which was assumed to consist of electrostatic, van der Waals, and depletion components. The results indicate that secondary energy wells significantly affect particle transport behavior through porous media. PMID- 16256618 TI - Analysis of the structure of very large bacterial aggregates by small-angle multiple light scattering and confocal image analysis. AB - This work aims at developing a more accurate measurement of the physical parameters of fractal dimension and the size distribution of large fractal aggregates by small-angle light scattering. The theory of multiple scattering has been of particular interest in the case of fractal aggregates for which Rayleigh theory is no longer valid. The introduction of multiple scattering theory into the interpretation of scattering by large bacterial aggregates has been used to calculate the fractal dimension and size distribution. The fractal dimension is calculated from the form factor F(q) at large scattering angles. At large angles the fractal dimension can also be computed by considering only the influence of the very local environment on the optical contrast around a subunit. The fractal dimensions of E. coli strains flocculated with two different cationic polymers have been computed by two techniques: static light scattering and confocal image analysis. The fractal dimensions calculated with both techniques at different flocculation times are very similar: between 1.90 and 2.19. The comparison between two completely independent techniques confirms the theoretical approach of multiple scattering of large flocs using the Mie theory. Size distributions have been calculated from light-scattering data taking into account the linear independence of the structure factor S(q) relative to each size class and using the fractal dimension measured from F(q) in the large-angle range or from confocal image analysis. The results are very different from calculations made using hard-sphere particle models. The size distribution is displaced toward the larger sizes when multiple scattering is considered. Using this new approach to the analysis of very large fractal aggregates by static light multiple scattering, the fractal dimension and size distribution can be calculated using two independent parts of the scattering curve. PMID- 16256619 TI - Behaviors of bovine serum albumin and rapeseed proteins at the air/water interface after grafting aliphatic or aromatic chains. AB - The influence of grafting aliphatic or aromatic groups on the behaviors of bovine serum albumin (BSA) and rapeseed proteins (napin and cruciferin) at the air/water interface is studied. From compression isotherms, it is shown that the chemical modification induces an increase in the interfacial molecular areas of the three proteins. The more hydrophobic the groups grafted, the more important this increase is. The dilatational modulus clearly emphasized that the grafting of hydrophobic groups also leads to an increase of the collapse pressure, demonstrating a higher cohesiveness and resistance to pressure of the interfacial films. These results are discussed on the basis of the physicochemical changes due to these chemical modifications, especially the conformation, the surface hydrophobicity, and the flexibility of the modified proteins. The improvement of surface properties obtained by grafting aliphatic or aromatic chains onto these proteins looks very promising in regard to emulsifying and foaming properties. PMID- 16256621 TI - Small-angle neutron scattering study of particle coalescence and SDS desorption during film formation from carboxylated acrylic latices. AB - Four monodisperse core-shell latices were synthesized for small-angle neutron scattering (SANS) studies, differing by the acrylic acid content in the particle shell (1 or 4 wt%) and the T(g) of the acrylic core (around -40 or 10 degrees C). In a first part, the coalescence kinetics of the surfactant-free latices were studied. It was shown that coalescence was hindered by an increase in the acrylic acid content of the shell, pH of the latex, and Tg of the core. These results could be interpreted in terms of chain mobility in the shell and in the core. Upon coalescence, the hydrophilic phase was segregated in spherical, polydisperse domains with an average diameter of 110 nm. In a second part, labeled SDS was used to follow desorption of the surfactant during film formation. It was shown that desorption occurred early in the film formation process when the latex still contained around 20% of water. A small fraction of the surfactant remained irreversibly adsorbed at the particle surface. PMID- 16256620 TI - A systematic study of bovine serum albumin (BSA) and sodium dodecyl sulfate (SDS) interactions by surface tension and small angle X-ray scattering. AB - Classical parameters obtained from surface tension technique coupled to small angle X-ray scattering (SAXS) measurements gave support to investigate conformational changes in the bovine serum albumin (BSA)-sodium dodecyl sulfate (SDS) complexes, as well as the size of the micelle-like clusters distributed along the polypeptide chain. The studied systems were composed of 1 wt% of BSA in the absence and presence of increasing SDS molar concentration up to 80 mM, under experimental conditions of low ionic strength and pH 5.40. At SDS concentrations below the critical aggregation concentration (cac) of 2.2 mM, SAXS results indicate that the detergent does not modify the native protein conformation. However, the beginning of protein unfolding, evidenced by SAXS through an increase in the values of radius of gyration Rg and protein maximum dimension Dmax, is coincident with the onset of SDS cooperative binding to BSA identified by the first breakpoint in the surface tension-SDS profile. Further SDS addition leads to the formation of micelle-like aggregates randomly distributed along the unfolded polypeptide chain, consistent to a necklace and bead model. The SAXS data also demonstrate that the SDS micelles grow in size up to 50 mM detergent. At 50 mM surfactant, the micelles stop growing. This concentration is near the BSA saturation binding by SDS measured by dialyzes and indicated by the second breakpoint in surface tension-SDS profile. The SAXS and surface tension data are also consistent with the formation of free micelles in equilibrium with BSA-SDS complexes for surfactant amount above the saturation. PMID- 16256622 TI - Preparation and characterization of polypyrrole-silica colloidal nanocomposites in water-methanol mixtures. AB - The effect of methanol cosolvent on the synthesis of polypyrrole-silica colloidal nanocomposites using ultrafine silica sols in combination with both FeCl3 and APS oxidants has been investigated. Two protocols were evaluated: the addition of methanol to an aqueous silica sol and the addition of water to a methanolic silica sol. The latter protocol proved to be more robust, since it allowed colloidally stable dispersions to be prepared at higher methanol content (up to 50 vol% using the APS oxidant). This allowed greater control over the particle size of the nanocomposite particles. In general, the spectroscopic data, the particle size range, silica contents and electrical conductivities of these nanocomposites were similar to those reported earlier for purely aqueous formulations. Polypyrrole contents ranged from 49 to 71% by mass and particle diameters varied from around 160 to 360 nm. In terms of colloid stability, the APS oxidant was preferred for nanocomposite syntheses in the presence of methanol. However, the FeCl3 oxidant generally gave higher conductivities and narrower size distributions under comparable conditions. HF etching experiments combined with transmission electron microscopy studies indicated that, to a first approximation, these nanocomposite particles had core-shell morphologies, with a hydrophobic polypyrrole core and a hydrophilic silica shell that compose approximately one monolayer of silica sol particles. Finally, aqueous electrophoresis measurements suggested that the polypyrrole-silica nanocomposites were silica-rich and that the methanolic silica sol was more hydrophobic (lower surface charge density) than the aqueous silica sol. PMID- 16256623 TI - Fabrication and surface characterization of NH4PAA-stabilized HAZ suspensions. AB - Fine composite powders of yttria (3 mol%) stabilized zirconia (Z, 10 wt%) and hydroxyapatite (HA), denoted as HAZ, were prepared by the co-precipitation method. The resulting powders were characterized by XRF, TEM, EDS, XRD, FTIR, TG DTA, and BET surface area techniques. AES and FTIR were employed to determine the surface properties of the HAZ suspensions in the presence of NH4PAA as a dispersant, which confirmed that the surfaces of both HA and Z were affected by the adsorbed polymers. The mechanism of NH4PAA adsorption on the particles was discussed. Zeta potential measurements showed that the addition of NH4PAA resulted in a dramatic increase in the absolute value of zeta potential. NH4PAA considerably enhanced the stability of the HAZ suspension via electrosteric barrier mechanisms. TEM micrographs confirmed that particles were well dispersed in the suspension. The adsorption density of the dispersant was found to decrease with an increase in pH value. PMID- 16256624 TI - Spreading behavior of crude oil over limestone substrate. AB - Wetting characteristics of crude oil droplets over limestone substrates in the presence of different aqueous solutions are investigated in terms of wetting length, droplet depth, contact angle, and spreading coefficient. A wide range of concentration of NaOH, Alcoflood polymer, and nonionic Triton X-100 surfactant are used as a continuous phase for the crude oil droplet. NaOH and Triton X-100 significantly enhanced the spreading characteristics of the crude oil droplet; however, AF1235 polymer caused a huge reduction in the spreading behavior of crude oil. PMID- 16256625 TI - Evaluation of DLVO theory with disjoining-pressure and film-conductance measurements of common-black films stabilized with sodium dodecyl sulfate. AB - We develop a unique film holder combining a thin-film balance with AC impedance spectroscopy to measure disjoining pressure, film conductance, and film thickness simultaneously. Foam films stabilized by sodium dodecyl sulfate (SDS) are investigated with and without added sodium chloride (NaCl) electrolyte. Classical colloidal theory, Derjaguin, Landau, Verwey, and Overbeek (DLVO) theory, is tested rigorously over a wide range of solution conditions by comparing the surface charge densities fit to disjoining-pressure isotherms with those estimated independently from film-conductance and surface-tension data. Film conductance measurements strongly suggest that the adsorbed anionic surfactant is partially complexed with counterions. Therefore, to reconcile the different values of charge densities calculated from surface tension and film conductance with those from disjoining pressure, we propose a simple ion-binding electrostatic model. The ion-complexation framework predicts increased ion complexing with increasing solution ionic strength, in agreement with surface tension and film-conductance data. Unfortunately, it is not possible to describe similarly the trends of the measured disjoining-pressure isotherms because the diffuse-layer charge density increases, or equivalently, the ion complexation decreases with increasing ionic strength. Accordingly, the ion-binding extension of classical DLVO theory does not permit agreement between theory and independent experimental data from surface tension, disjoining pressure, and film conductance. PMID- 16256627 TI - Vacuum removal of the template in MCM-41 silica studied by the positron annihilation method. AB - Positron annihilation lifetime spectra were measured for ordered silica before template (octadecyltrimethyl ammonium) removal. An intermediate lifetime component, well pronounced in the spectra, indicates the presence of structural defects in the template being the size of a surfactant molecule. At the temperature near 420 K a radical change of template structure occurs, consisting in appearance of disorder and the degradation of template; evaporation of the fragments begins. It seems that moderate heating (up to about 500 K only) in vacuum for several hours is an effective and simple method of template removal, leaving no pure carbon residue in the pore structure. The results are compared with the data from the liquid nitrogen adsorption/desorption method and elemental analysis, confirming the positron annihilation results. PMID- 16256626 TI - Self-assembly and morphology of gel networks in l,3:2,4-bis-O-(p methylbenzylidene)-D-sorbitol/n-dibutylphthalate. AB - We investigated the self-assembling structure of the 1,3:2,4-bis-O-(p methylbenzylidene)-D-sorbitol (PDTS)/n-dibutylphthalate (DBP) system in the parameter space of temperature T and solute concentration Phi(PDTS). Optical microscopic studies revealed that the phase diagram can be divided into four regions. In region I at high T the system is a homogeneous solution. In region II at lower T and low Phi(PDTS), the system still has fluidity but has microgels having spherulitic texture of PDTS crystallites. Regions III and IV at even lower T but higher Phi(PDTS) are in a gel state. In region III, the PDTS forms volume filling spherulites due to the solid-liquid phase transition of the saturated PDTS solutions. In region IV at the lowest T, both the liquid-liquid phase separation process and the solid-liquid transition of the PDTS are involved in the self-assembling processes. In this region a bicontinuous phase-separated structure is first formed by liquid-liquid phase separation via spinodal decomposition (SD). The subsequent solid-liquid transition of the PDTS in the PDTS-rich region forms percolating crystalline fibrils rather than spherulites. The formation of the crystalline fibrils pins further growth of the bicontinuous structure via SD. PMID- 16256628 TI - Determination of gas-oil miscibility conditions by interfacial tension measurements. AB - Processes that inject gases such as carbon dioxide and natural gas have long been and still continue to be used for recovering crude oil from petroleum reservoirs. It is well known that the interfacial tension between the injected gas and the crude oil has a major influence on the efficiency of displacement of oil by gas. When the injected gas becomes miscible with the crude oil, which means that there is no interface between the injected and displaced phases or the interfacial tension between them is zero, the oil is displaced with maximum efficiency, resulting in high recoveries. This paper presents experimental measurements of interfacial tension between crude oil and natural gases (using a computerized drop shape analysis technique) as a function of pressure and gas composition at the temperature of the reservoir from which the crude oil was obtained. The point of zero interfacial tension was then identified from these measurements by extrapolation of data to determine minimum miscibility pressure (MMP) and minimum miscibility composition (MMC). The gas-oil miscibility conditions thus obtained from interfacial tension measurements have been compared with the more conventional techniques using slim-tube tests and rising-bubble apparatus as well as predictive correlations and visual observations. The miscibility pressures obtained from the new VIT technique were 3-5% higher than those from visual observations and agreed well with the slim-tube results as well as with the correlations at enrichment levels greater than 30 mol% C2+ in the injected gas stream. The rising bubble apparatus yielded significantly higher MMPs. This study demonstrates that the VIT technique is rapid, reproducible, and quantitative, in addition to providing visual evidence of gas-oil miscibility. PMID- 16256629 TI - An improved estimation of water-organic liquid interfacial tension based on linear solvation energy relationship approach. AB - An equation based on the linear solvation energy relationship (LSER) was proposed to predict the interfacial tension between organic liquid and water. The equation takes into account five parameters characterizing properties of the organic liquid molecule: excess molar refraction, solute dipolarity/polarizability, effective hydrogen bond acidity, effective hydrogen bond basicity, and the McGowan molar intrinsic volume. The proposed equation provides a better approximation of the interfacial tension than a similar one derived earlier by Freitas et al. (J. Phys. Chem. B 101 (1997, 7488-7493), which is based on seven terms. PMID- 16256630 TI - Validity and accuracy in evaluating surface tension of solids by additive approaches. AB - The validity and the accuracy of both the Owens and Wendt and the Lifshitz-van der Waals/acid-base (LW/AB) methods for the determination of surface tensions of solids have been examined for a wide variety of situations. In each case, the allowed range of contact angles that result in positive values of all the square roots of the surface tension components of the solid has first been determined. Then the maximum relative errors in the surface tensions of solids that result from errors in contact angle measurements have been calculated within the allowed range. For both methods, it has been found that the maximum relative errors are minimal if one of the liquids is apolar. In the case of the LW/AB method, minimal errors are obtained if, in addition, the other two liquids are monopolar with different polarities. However, the more similar are the properties of the liquids, the narrower is the allowed range, and the larger are the maximum relative errors. PMID- 16256631 TI - Effect of ionic surfactants on the phase behavior and structure of sucrose ester/water/oil systems. AB - The phase behavior and structure of sucrose ester/water/oil systems in the presence of long-chain cosurfactant (monolaurin) and small amounts of ionic surfactants was investigated by phase study and small angle X-ray scattering. In a water/sucrose ester/monolaurin/decane system at 27 degrees C, instead of a three-phase microemulsion, lamellar liquid crystals are formed in the dilute region. Unlike other systems in the presence of alcohol as cosurfactant, the HLB composition does not change with dilution, since monolaurin adsorbs almost completely in the interface. The addition of small amounts of ionic surfactant, regardless of the counterion, increases the solubilization of water in W/O microemulsions. The solubilization on oil in O/W microemulsions is not much affected, but structuring is induced and a viscous isotropic phase is formed. At high ionic surfactant concentrations, the single-phase microemulsion disappears and liquid crystals are favored. PMID- 16256632 TI - An investigation of dynamic surface tension, critical micelle concentration, and aggregation number of three nonionic surfactants using NMR, time-resolved fluorescence quenching, and maximum bubble pressure tensiometry. AB - Several physicochemical properties have been determined for N dodecyllactobionamide (LABA), maltose 6'-O-dodecanoate (C12-maltose ester), and tetra(ethylene oxide) dodecyl amide (TEDAd). The increase in the flexibility of the sugar headgroup, enabling more possible molecular conformations, reduces the minimum area/molecule at the liquid-vapor interface obtained at the critical micelle concentration (cmc). The obtained cmc's were 0.35 mM (LABA), 0.3 mM (C12 maltose ester), and 0.5 mM (TEDAd). The monomer diffusion coefficient decreased with the molecular weight and increasing headgroup flexibility of the sugar headgroup, and values were in the range from 3.1 x 10(-10) to 3.6 x 10(-10) m2/s. The micelle diffusion coefficients (0.46 x 10(-10) to 0.68 x 10(-10) m2/s) indicated that the TEDAd micelles deviated most from spherical shape. The micelle aggregation numbers determined by time-resolved fluorescence quenching (TRFQ) were estimated to be 120+/-10 (LABA), 90+/-10 (C12-maltose ester), and 130+/-10 (TEDAd). The dynamic surface tension measurements show that the adsorption of TEDAd onto the liquid-vapor interface at short surface lifetimes is diffusion limited, whereas an adsorption barrier is present for the sugar surfactants. The analysis of the dynamic surface tension data above the cmc shows that the rate of demicellization is faster for TEDAd than for the two sugar-based surfactants. PMID- 16256633 TI - Structural effects on surface and micellar properties of alkanediyl-alpha, omega bis(sodium N-acyl-beta-alaninate) gemini surfactants. AB - A study of the equilibrium surface properties (in water and in the presence of 0.01 M NaCl) of a novel series of anionic gemini surfactants, alkanediyl alpha,omega-bis(sodium N-acyl-beta-alaninates), is described. Parameters studied include cmc (critical micelle concentration), C20 (required to reduce the surface tension of the solvent by 20 mN/m), gamma(cmc) (the surface tension at the cmc), Gamma(max) (the maximum surface excess concentration at the air/aqueous solution interface), Amin (the minimum area per surfactant molecule at the air/water interface), and the cmc/C20 ratio (a measure of the tendency to form micelles relative to adsorb at the air/water interface). The geminis with a spacer consisting of two methylene groups show premicellar self-aggregation, both in water and in 0.01 M NaCl, when the N-acyl group contains more than 12 carbon atoms; geminis with a spacer consisting of four methylene groups show no premicellar aggregation even when the N-acyl group contains 16 carbon atoms. For the acyl chain lengths where premicellar aggregation does not occur, the values of the cmc of the geminis with a two-methylene spacer are lower than those for the corresponding analogous geminis with a four-methylene spacer. The premicellar formation for the geminis with a two-methylene spacer is due to the short-chain linkage. The geminis show little or no break in their specific conductance surfactant molar concentration plots and an increase in the pH at the cmc. This is attributed to protonation of the carboxylate group and strong Na+ release during micellization. PMID- 16256634 TI - Micellar behavior of tetradecyldimethylbenzylammonium chloride in water-alcohol mixtures. AB - The effect of butanol and benzyl alcohol on the critical micelle concentration and the degree of ionization of micelles of tetradecyldimethylbenzylammonium chloride has been studied conductometrically in the temperature range of 5 to 40 degrees C at 5 degrees C intervals. The results indicate that some self association process of benzyl alcohol in the aqueous phase commences when its concentration amounts to ca 0.05 m. By applying the theoretical treatment suggested by Motomura for binary surfactant systems, the molar fraction of alcohol in the micelle and the standard free energy of solubilization were estimated from the dependence of cmc on temperature and alcohol concentration. For comparative purposes the micellar properties of tetradecyltrimethylammonium bromide in water-alcohol systems at 25 degrees C have also been studied. PMID- 16256635 TI - Effects of bile salts on percolation and size of AOT reversed micelles. AB - The effects of two trihydroxy bile salts, sodium taurocholate (NaTC) and 3-[(3 cholamidylpropyl)dimethylammonio]-1-propane sulfonate (CHAPS), on the size, shape and percolation temperature of reversed micelles formed by sodium bis(2 ethylhexyl)sulfosuccinate (AOT) in isooctane were studied. The percolation temperature of the reversed micelles decreased upon inclusion of bile salts, indicating increased water uptake. Dynamic light scattering (DLS) measurements showed consistent enlargement of reversed micelles upon addition of the bile salts; the hydrodynamic radius increased sixfold in the presence of 10 mM CHAPS and doubled in the presence of 5 mM NaTC. Inclusion of the enzyme yeast alcohol dehydrogenase (YADH) increased the percolation temperature and distorted the spherical structure of the AOT reversed micelles. The spherical structure was restored upon addition of bile salt. These results may help to explain the increase in activity of YADH in AOT reversed micelles upon addition of bile salts. PMID- 16256637 TI - Poly(butyl methacrylate-g-methoxypoly(ethylene glycol)) and poly(methyl methacrylate-g-methoxypoly(ethylene glycol)) graft copolymers: preparation and aqueous solution properties. AB - A series of water-soluble, amphiphilic graft copolymers has been prepared by free radical copolymerization of methoxypoly(ethylene glycol) macromonomers, with either methyl methacrylate or butyl methacrylate as the comonomers, in water/ethanol solvent mixtures. Lower molecular weight copolymers were obtained by increasing the concentration of the initiator, azobisisobutyronitrile (AIBN), used in the polymerization reaction. However, the route used also led to the formation of significant quantities of tetramethylsuccinodinitrile, a toxic byproduct resulting from the cage reaction of AIBN. Static fluorescence measurements using pyrene as a probe, along with 1H NMR experiments, showed that the graft copolymers form aggregates in water at very low concentrations (approximately 0.01 g l(-1)) with the pendant hydrophilic graft chains forming a stabilizing shell around the hydrophobic backbone. An increase in the hydrophile lipophile balance of the graft copolymers was found to lead to smaller aggregates with lower aggregation numbers and highly swollen hydrophilic shells, as revealed by small angle neutron scattering (SANS). PMID- 16256636 TI - Polystyrene nanoparticles based on poly(butyl methacrylate-g-methoxypoly(ethylene glycol)) and poly(methyl methacrylate-g-methoxypoly(ethylene glycol)) graft copolymers. AB - The solubilization of styrene by poly(butyl methacrylate-g-methoxypoly(ethylene glycol)) and poly(methyl methacrylate-g-methoxypoly(ethylene glycol)) graft copolymers has been examined. From turbidity measurements the solubility limit of the monomer in the micelles was obtained and the distribution coefficients were evaluated. Dynamic light scattering revealed that below the solubility limit, solubilization leads to a slight increase in micelle size, while above the solubility limit, there is a dramatic increase in particle size and turbidity as oil-in-water emulsions are formed through coalescence of monomer-swollen micelles. Polymerizations carried out below the solubility limit using the graft copolymer micelles as templates resembled microemulsion polymerizations in nature and led to very fine sterically stabilized polystyrene latex particles. Through careful control of the monomer concentration and the polymerization temperature it was possible to obtain spherical nanosize latex particles with similar size to those of the micelle precursors (10 nm) up to 11% monomer by weight. Polymerizations above the solubility limit, on the other hand, showed similarities with emulsion polymerizations and resulted in larger particles with higher polydispersity. PMID- 16256638 TI - Probing the association behavior of poly(ethylene glycol)-based amphiphilic comb like polymer in NaCl solution. AB - The effect of salt on the associative behavior of intramolecular aggregates obtained from poly(ethylene glycol)-based amphiphilic comb-like polymers in aqueous medium at pH 6.2 has been investigated by surface tension, fluorescence probe, dynamic light-scattering, and viscometry techniques. Results reveal that the addition of salt screens the electrostatic repulsion between the charges along the polymer backbone in the aggregates and consequently (1) reduces the surface activity at the air/water interface, (2) leads to the contraction of the polymer backbone, and (3) reduces the hydrodynamic sizes of the aggregates. In contrast, the hydrophobicity of the aggregates remains unperturbed. PMID- 16256639 TI - Studies on the electrochemical and permeation characteristics of asymmetric charged porous membranes. AB - Asymmetric charged porous membranes were prepared by imbedding 10% (W/W) ion exchange resin in cellulose acetate binder. Membrane potential and conductance measurements have been carried out in sodium chloride solutions at different concentrations to investigate the relationship between concentration of fixed charges and electrochemical properties of developed nonselective cation- and anion-exchange membranes. Counterion transport number and permselectivity of these membranes were found to vary due to the presence of ion-exchange resin. The hydrodynamic and electroosmotic permeability of sodium chloride solutions has been studied in order to compute equivalent pore radius. For cation- and anion exchange membranes good agreement was observed between pore radius values estimated from hydrodynamic and electroosmotic permeability coefficient separately, while for nonselective membranes no correlation was found. Membrane conductance data, along with values of concentration of fixed charges, were used for the estimation of the tortuosity factor, salt permeability coefficient, and frictional coefficient between solute and membrane matrix employing an interpretation by nonequilibrium thermodynamic principles based on frictional forces. Moreover, surface morphological studies of these membranes also have been carried out and the membranes were found to be reasonably homogeneous. PMID- 16256640 TI - Confinement of polyaniline chains in capillary layers: an ordering effect of the uniaxially aligned surfaces. AB - An ordering effect of uniaxially aligned poly(tetrafluoroethylene) (PTFE) substrates prepared by rubbing on polyaniline (PANI) molecules at the interfaces of PTFE/PANI film and PTFE/PANI solution has been investigated using electronic absorption spectroscopy. It was observed slight dichroism in electronic spectra from only very thin (thickness approximately 20 nm and less) PANI films as well as from PANI solutions of capillary thickness (10 to 30 microm) confined by oriented PTFE surfaces. The ordering effect is discussed in terms of a hydrodynamic flow arising upon sample formation and steric factors at the PTFE surface, which cause uniaxial deformation of the polymer coil on the rubbed PTFE surface. PMID- 16256641 TI - Characterization of the solvation environment provided by dilute aqueous solutions of novel siloxane polysoaps using the fluorescence probe pyrene. AB - Solubilization environment afforded by several of the novel allyl glycidyl ether modified methylhydrosiloxane polymers are investigated using a common polycyclic aromatic hydrocarbon fluorescence probe, pyrene. The backbone of the polymer has been modified by the addition of an alkyl chain of varying length (either C8, C12, or C18) and to differing degrees of substitution. The nomenclature adopted for the purposes of these studies is as follows: "AGENT" represents the backbone polymer with no alkyl substitution, and "OAGENT", "DAGENT", and "SAGENT" are substituted with n-octyl, n-dodecyl, and n-octadecyl, respectively. The percentage of alkyl substitution is designated as 10, 15, and 20%. The pyrene polarity scale (defined as the ratio of the intensity of peak I to peak III) was used to determine the relative dipolarity of the cybotactic region provided by approximately 1 w/w% aqueous polymer solutions compared to 10 mM sodium dodecylsulfate (SDS) micellar solution. Results indicate that 10-15% DAGENT afforded the most hydrophobic solubilization site, followed by 15% OAGENT and 15% SAGENT. In addition, as the degree of alkyl substitution of DAGENT increased from 10 to 20%, the cybotactic region appeared to become more hydrophobic. Furthermore, a deeper investigation into the relative size of the solubilization site revealed that all alkyl-substituted polymers promoted excimer formation at relatively low pyrene concentrations, indicating the possibility of localized concentration enhancement within the solvation pockets and/or compartmentalization of the solute molecules. The pyrene fluorescence excitation data strongly indicates ground-state heterogeneity that is most prominent in AGENT and decreases as the alkyl chain length is increased. This provides a relative sense of the size and shape of the solvation pockets afforded by each polymer solution. An overall analysis of the collected data indicated that these alkyl-substituted polymers may provide a more selective and efficient pseudostationary phase in electrokinetic chromatography with better solvation capacity for hydrophobic compounds compared to SDS. PMID- 16256642 TI - Optical properties of nanocrystalline Y2O3:Eu depending on its odd structure. AB - The structure of nanocrystalline Y2O3:Eu prepared by a combustion reaction was analyzed by XRD and high-resolution electron microscopy. Compared with a large scale particles, 5-nm Y2O3:Eu particles presented as distorted crystallite and rough surfaces. Luminescent and absorption properties of nano-Y2O3:Eu showed remarkably particle size effects. At Y2O3:Eu particle sizes smaller than 10 nm some new results were observed: (a) a red shift of the charge-transfer-state absorption; (b) new emission bands of Eu3+ in the 5D0 --> 7F2 region; (c) luminescent decay of energy level 5D0 of Eu3+ turning to a two-step exponential; and (d) a pronounced increase in quenching concentration and much lower phonon density compared with those of the bulk material. All these phenomena can be attributed to the effect of the softened lattice and surface state of the nanomaterial. The latter was confirmed by stronger excitation by the host absorption after the surface modification. PMID- 16256643 TI - Interpreting the signal from a localized fluorescence sensor: a study by angle resolved XPS and dynamic SIMS. AB - Angle-resolved X-ray photoelectron spectroscopy (XPS) and dynamic secondary ion mass spectroscopy (DSIMS) experiments were conducted to assess the interactions between a diamine curing agent and a glycidoxysilane-modified glass substrate. This effort was motivated by earlier work, in which a fluorescent probe localized in dilute quantities in the silane layer was used to track the penetration of the resin into the silane layer, as well as the resin cure. XPS and DSIMS experiments were performed on the silane layers immersed only in the resin hardener, providing more detailed information about the concentration profile and structural reorganization within the silane layer due specifically to hardener penetration. Dynamic SIMS spectra reveal the presence of hardener in the layer, as indicated by the strong CN- signal throughout the silane layer thickness. The XPS results indicate the presence of an amine gradient within the top 10 nm of the silane coating, with less amine penetration deeper into the silane layer. The XPS data also suggest some level of anisotropy in the molecular structure of the diamine/glycidoxysilane coating, as revealed by the differences in the relative atomic concentrations and peak positions of the C1s components at two different take-off angles. PMID- 16256644 TI - Extension of the Dubinin-Astakhov equation for evaluating the micropore size distribution of a modified carbon molecular sieve. AB - A new method for the characterization of the pore size distribution of microporous solids is applied on data obtained for activated carbon molecular sieve samples. In this method, based on the Dubinin-Astakhov equation, a simple numerical algorithm is used for the reconstruction of the micropore size distribution from the integral equation that represents the experimental nitrogen adsorption isotherm. The results are compared with the ones obtained on the basis of the well-known Horvath-Kawazoe method. The samples used in this study come from a carbon molecular sieve that has been treated with solutions of concentrated HNO3 at various temperatures and with solutions of H2O2 of various concentrations. PMID- 16256645 TI - Role of surface chemistry in adsorption of phenol on activated carbons. AB - Two samples of activated carbon of wood origin were oxidized using ammonium persulfate. The structural properties and surface chemistry of the samples and their oxidized counterparts were characterized using sorption of nitrogen and Boehm titration, respectively. Phenol adsorption from solution (at trace concentrations) was studied at temperatures close to ambient without maintaining a specific pH of the solution. The results showed, as expected, that the phenol uptake is dependent on both the porosity and surface chemistry of the carbons. Furthermore, phenol adsorption showed a strong dependence on the number of carboxylic groups due to two factors: (1) phenol reacts with carboxylic groups on the carbon surface, forming an ester bond, and (2) carboxylic groups on the carbon surface remove the pi-electron from the activated carbon aromatic ring matrix, causing a decrease in the strength of interactions between the benzene ring of phenol and the carbon's basal planes, which decreases the uptake of phenol. PMID- 16256646 TI - Molecular weight fractionation of humic substances by adsorption onto minerals. AB - Molecular weight (MW) fractionation of Suwannee River fulvic acid (SRFA) and purified Aldrich humic acid (PAHA) by adsorption onto kaolinite and hematite was investigated in equilibrium and rate experiments with a size-exclusion chromatography system using ultraviolet (UV) light detection. The extent of adsorptive fractionation based on UV detection was positively correlated with the percent carbon adsorption for both humic substances (HS), although the specific fractionation pattern observed depended on the particular HS and mineral used. Higher MW fractions of SRFA, an aquatic HS, were preferentially adsorbed to both kaolinite and hematite whereas the fractionation trends for PAHA, a terrestrial peat HS, differed for the two minerals. The contrasting fractionation patterns for SRFA versus PAHA can be explained reasonably well by the different structural trends that occur in their respective MW fractions and the underlying adsorption processes. Rate studies of adsorptive fractionation revealed an initial rapid uptake of smaller HS molecules by the mineral surfaces, followed by their replacement at the surface by a much slower uptake of the larger HS molecules present in aqueous solution. PMID- 16256647 TI - Effect of low-molecular-weight organic anions on surface charge of variable charge soils. AB - Low-molecular-weight (LMW) organic acids exist widely in soils and have been implicated in many soil processes. In the present paper, the effect of the anions of four low-molecular-weight organic acids on the surface charge of three variable charge soils was investigated. The results showed that the presence of organic anions led to an increase in negative charge and a decrease in positive charge. Positive charges decreased to a larger extent than negative charges. The effect of different anions on surface charge followed the order citrate > malate > oxalate > acetate. For hyper-rhodic ferrasol and rhodic ferrasol, the change of positive charge decreased with the increase in pH, while that of negative charge increased with the increase in pH. Among different soils the extent of change in surface charge was related to their iron oxides content. When free iron oxides were removed from the soil, the effect of organic anions on surface charge decreased sharply. These findings may be of practical significance for variable charge soils low in nutrient-retaining capacity by increasing the retaining capacity for cations such as potassium and calcium considerably but decreasing that for anions such as nitrate remarkably. PMID- 16256648 TI - Adsorption of arsenate from water using neutralized red mud. AB - The potential for using seawater-neutralized red mud (Bauxsol), a waste from aluminum manufacturing, as an adsorbent for removing As(V) (arsenate) from water is studied. Herein, adsorption characteristics are investigated and it is shown that adsorption follows the Langmuir model, with the adsorption constants indicating the feasibility of the process. Furthermore, the adsorption is found to increase with decreasing pH (i.e., ligand-like adsorption), higher adsorbent dosages, and lower initial arsenate concentrations. The effects of ions are also tested and it is shown that the adsorption of arsenate decreases in the presence of HCO3-, while Cl- has little effect, and Ca2+ increases the adsorption. Water quality assessment after treatment with Bauxsol indicates that none of the trace elements tested are released from the adsorbent. A TCLP leaching test also reveals that the used adsorbent is not toxic. It is foreseen that Bauxsol may be developed into an efficient low-cost adsorbent for (pre-) treating arsenate contaminated waters. PMID- 16256649 TI - Analytical investigation of two-step adsorption kinetics on surfaces. AB - Analytical equations of two-step adsorption kinetics on surface have been derived. Moreover, computer simulations have been carried out to interpret various experimental adsorption kinetics previously reported. In the first case, molecules are further adsorbed from a solution onto a layer consisting of previously adsorbed molecules. This model was applied to the adsorption kinetics of hexadecyltrimethylammonium chloride (C16TAC) on a self-assembled monolayer (SAM) of 3-mercaptopropionic acid (T. Imae, H. Torii, J. Phys. Chem. B 104 (2000) 9218). The second case is that some of the initially adsorbed molecules are released from the adlayer with further time course. The adsorption of C16TAC on 1 dodecanethiol SAM (T. Imae, T. Takeshita, K. Yahagi, Stud. Surf. Sci. Catal. 132 (2001) 477) agrees with this mechanism. The strict mathematical developments presented in this work are demanded to specify the physical meaning of observed non-Langmuir adsorption kinetics, consisting of the two exponential terms. PMID- 16256650 TI - Structural-chemical disorder of manganese dioxides II. Influence on textural properties. AB - Relationships between structural parameters of MnO2 and their surface properties at the solid-gas interface were investigated. The studied series ranged from ramsdellite to pyrolusite and encompassed disordered gamma-MnO2 samples. The structural model used takes into account structural defects: Pr (rate of pyrolusite intergrowth in the ramsdellite network) and Tw (rate of microtwinning). Analysis of the N2 adsorption isotherm evidenced positive correlations between specific surface area and Tw for gamma-MnO2 only and between the energetic constant C and (1-Pr). No microporosity is evidenced. Water adsorption isotherms evidenced the dependence of the H2O monolayer volume on Tw and showed a positive correlation between the cross-section area of water molecules adsorbed in the first monolayer and Pr, ranging from 13.5 A2 for Pr=1 to 6.3 A2 for Pr=0.2 (12 sites/nm2). Energetic heterogeneity is quantified from Ar and N2 low-pressure adsorption isotherms with the DIS procedure and correlated with H2O adsorption. High-energy adsorption domains are quantified and assigned to the different crystal faces: (110) faces with a common 1 x 1 octahedra layer of pyrolusite and ramsdellite and the (001) face of ramsdellite with 2 x 2 octahedra on which channels and plateaus are differentiated. The specific surface area ratio of ramsdellite high-energy sites to total ramsdellite content is shown to depend on Tw. The dependence on microtwinning of low cross-sectional area of N2 and much lower cross-sectional of residual H2O molecules leads us to assume that their adsorption sites on grain boundaries are represented by the twin planes between the structured nanocrystals generated by oxygen evolution during MD synthesis. PMID- 16256651 TI - Surface hydroxylation and silane grafting on fumed and thermal silica. AB - The optimization of the surface functionalization of flat thermal silicon oxide by silanes was investigated. The difficulties are the low density of silanols at the surface of thermal silica, the lack of precise knowledge of the actual surface chemistry of thermal silica and of its hydroxylation, and the limited number of possible chemical analyses at flat surfaces of small area. This steered our study toward a comparative investigation of the hydroxylation and silane grafting of thermal silica and the well-known fumed silica. The silane grafting density for fumed silica that had undergone thermal treatments of dehydroxylation was related to the surface density of silanols. The surface density of silane on the flat thermal silica as measured by FTIR-ATR spectroscopy was 1.4 micromol/m2, similar to that of fumed silica dehydroxylated at 1000 degrees C. This moderate value was related to the low silanol density present on such silica surfaces. Several rehydroxylation treatments that proved their efficiency on dehydroxylated fumed silica did not lead to any noticeable improvement on thermal silicon dioxide. PMID- 16256652 TI - Improved surface properties of polyaniline films by blending with Pluronic polymers without the modification of the other characteristics. AB - Films of conductive polyaniline and amphiphilic Pluronic (P105) copolymer blends were prepared by dissolving the two polymers in N-methylpyrrolidinone (NMP) followed by a slow solvent evaporation at 55 degrees C. The characteristics of both doped and undoped films were determined by X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM), water droplet contact angles, differential scanning calorimetry (DSC), thermal gravimetry analysis (TG), wide angle X-ray diffraction (WAXD), and tensile strength measurements. The surface of the blends became more hydrophilic than that of the hydrophobic PANI film, but the other properties of the blends did not change appreciably for Pluronic content lower than 50 wt%. Compared to PANI films, the more hydrophilic surfaces decreased the amount of bovine serum albumin protein adsorbed. By preventing biofouling, the polyaniline-Pluronic blends can become more useful as biosensors than the polyaniline films. PMID- 16256653 TI - Two liquid adsorptive entrapment of a pluronic polymer into the surface of polyaniline films. AB - Pluronic triblock copolymers were entrapped on the surface of polyaniline (PANI) films by first immersing the latter in N-methylpyrrolidinone (NMP) solutions of one of the Pluronics for a short time. This softened the surface of the films and allowed the Pluronic molecules to entangle with PANI segments of the swollen film on the surface. Further, the films were taken out from the NMP solution and dipped into water, which is a nonsolvent for PANI. The rapid surface deswelling of PANI by the water resulted in the entrapment of the Pluronic on its surface, with the hydrophilic blocks toward water and the hydrophobic block imbedded in the PANI films. The modified PANI obtained was examined by X-ray photoelectron spectroscopy, water droplets contact angles, scanning electron microscopy, and wide angle X-ray diffraction. The surface of the Pluronic entrapped PANI films became more hydrophilic than the hydrophobic PANI films and decreased the amount of bovine serum albumin protein adsorbed on them. This means that, by reducing the biofouling, the life of the modified polyaniline film can be extended when the latter is employed as a biosensor. PMID- 16256654 TI - Low-temperature synthesis of niobium oxide nanoparticles from peroxo niobic acid sol. AB - A peroxo niobic acid sol was prepared by peptization of the niobic acid precipitate (Nb2O5.nH2O) with a H2O2 aqueous solution. Crystallized Nb2O5 nanoparticles and niobic acid nanoparticles were obtained by heating the peroxo niobic acid sol. When peroxo niobic acid sol prepared by peptization of the niobic acid precipitate ([NH3]=0.3 mol/l) was heated at 348 K for 1 week, Nb2O5 nanoparticles with a diameter of 4.5 nm and a S(BET) of 275 m2/g were obtained. When peroxo niobic acid sol prepared by peptization of the niobic acid precipitate ([NH3]=1 mol/l) was heated at 348 K for 1 week, niobic acid nanoparticles with a diameter of less than 2 nm were obtained. The pore structure and degree of crystallinity of the nanoparticles prepared by heating the peroxo niobic acid sol greatly depended on the concentration of the ammonia solution used for preparing the niobic acid precipitate. PMID- 16256655 TI - Direct coating of gold nanoparticles with silica by a seeded polymerization technique. AB - Gold nanoparticles prepared through a conventional citrate-reduction method were directly coated with silica by means of a seeded polymerization technique based on the Stober method. The method required no surface modification. The addition of tetraethylorthosilicate and water prior to ammonia was found to be critical to obtain a proper coating. The silica shell thickness was varied from 30 to 90 nm for TEOS concentrations of 0.0005-0.02 M at 10.9 M of water and 0.4 M of ammonia. The optical spectra of the core-shell gold-silica composite particles agreed with predictions of Mie theory. PMID- 16256656 TI - Synthesis and assembly of SiO2-coated Bi2S3 nanofibers. AB - The SiO2 coating of Bi2S3 nanofibers using the alkaline hydrolysis of Si(OEt)4 is reported. A comparative study on the optical and morphological properties of the starting Bi2S3 nanofibers and the final SiO2 coated nanomaterials are presented and discussed. A SiO2 cap covering homogeneously single Bi2S3 nanofibers was obtained when the hydrolysis of Si(OEt)4 was performed under sonication treatment to maintain the fibers well separated during the coating process. These coated nanofibers were then used in a layer-by-layer deposition process to produce multilayered nanostructured films. PMID- 16256657 TI - Phase transfer of silver nanoparticles from aqueous to organic solutions using fatty amine molecules. AB - We demonstrate the phase transfer of silver nanoparticles synthesized in an aqueous medium into hexane containing the cationic surfactant octadecylamine (ODA). During vigorous shaking of the biphasic mixture, rapid phase transfer of the silver nanoparticles into the organic phase was observed. The phase transfer of the silver nanoparticles arises due to coupling of the silver nanoparticles with the ODA molecules present in organic phase via either coordination bond formation or weak covalent interaction. This process renders the nanoparticles sufficiently hydrophobic and dispersible in the organic phase. The ODA-stabilized silver nanoparticles could be separated out from the organic phase in the form of a powder and are readily redispersible in different organic solvents. The nature of binding of the ODA molecules to the silver nanoparticle surface was characterized using UV-vis spectroscopy, thermogravimetry, transmission electron microscopy, nuclear magnetic resonance spectroscopy, X-ray photoemission spectroscopy, and Fourier transform infrared spectroscopy. PMID- 16256658 TI - The importance of temperature and viscosity effects for surfactant adsorption measurements made using the electrochemical quartz crystal microbalance. AB - The measurement of adsorbed surfactant is important to fields such as corrosion inhibition, metal cleaning, and separation technologies. The electrochemical quartz crystal microbalance (EQCM) is an important tool that can be used to measure adsorbed surfactant. However, such measurements are subject to significant temperature and viscosity effects that must be appropriately considered. This paper discusses the effect of temperature and viscosity on EQCM measurements in solution environments and the ability of the EQCM to measure surfactant adsorption. PMID- 16256659 TI - On the adsorption and condensed film formation of dodecyl-, tetradecyl-, hexadecyl-, and octadecyltrimethylammonium bromides at the mercury/electrolyte interface. AB - The adsorption and condensed film formation of dodecyl (DTAB)-, tetradecyl (TTAB) , hexadecyl (CTAB)-, and octadecyl (OTAB)-trimethylammonium bromides on the hanging mercury electrode is studied in KBr as supporting electrolyte, at various temperatures from 5 to 45 degrees C. A condensed film is formed at negative potentials and at room temperature only in the presence of CTAB. The decrease of the temperature favors the formation of the condensed film. A transition temperature is observed for the film formation. Capacity-time curves at the potentials where the film is formed show a nucleation and growth mechanism, with induction time depending not only on the final potential but also on the initial potential range, although it is in the desorption region. In this temperature range no film is observed for DTAB and TTAB. However, the film is observed for OTAB, but only at higher temperatures, and is more easily formed with increasing temperature. The film is formed in a certain potential region and the nucleation rate increases while moving toward more negative potentials. Hysteresis phenomena are observed during changes of scan direction. The capacity vs time curves for OTAB, where condensed film is formed, are treated using an Avrami plot formulation and have been explained as progressive one-dimensional nucleation with a decrease of the nucleation rate during the overall film formation. The results show a marked effect of the chain length of the alkyl chain on the film formation. PMID- 16256660 TI - Role of titania incorporated on activated carbon cloth for capacitive deionization of NaCl solution. AB - Adsorption isotherms of NaCl on activated carbon cloth (ACC) and titania incorporated activated carbon cloth (Ti-ACC) under an electric field were investigated to deduce the role of titania in capacitive deionization (CDI) of NaCl. Electrosorption of NaCl on the ACC was significantly increased by titania incorporation, whereas its physical adsorption was considerably decreased, resulting in an improved performance of the Ti-ACC as a CDI electrode. Langmuir isotherms based on a localized and fixed amount of adsorption were suitable for the simulation of electrosorption and physical adsorption of ions on the ACC electrodes. The variances of q(m) and b of Langmuir isotherms with electric potential indicate increases in the number of ions per adsorption site and in electrosorption strength of ions by titania incorporation. A cyclic voltammetric study for ion adsorption on ACC electrodes confirms the reversibility between electrosorption and desorption of ions, regardless of titania incorporation. PMID- 16256661 TI - Formation of a percolating cluster in films prepared by cathodic electrodeposition of a mixture of lower and higher molecular weight epoxy-amine adducts. AB - The electrodeposition behavior of blends of primary dispersions of a lower and a higher molecular weight epoxy-amine adduct has been investigated. The throwing power of the above-mentioned blends showed a voltage-dependent critical composition at which the throwing power dropped to a much lower value. This was assigned to the formation of an infinite conducting cluster, the extension of which is dependent on the rate of the electrocoagulation process at the cathode boundary. The random resistor network approach of Stauffer (RRNS) and the random resistor network approach of Miller and Abrahams (RRNMA) were applied to the experimental data with high correlations (r2=0.9314 and 0.9699). The percolating cluster formed within the film, however, gave a critical exponent of conductivity equal to 1.1028, much less than expected from a classical three-dimensional lattice (i.e., 1.5-2.0). This discrepancy was explained in terms of the changed behavior of the film resulting from the bubbles formed near the cathode and its effect on the infinite conducting cluster. PMID- 16256663 TI - Laser excitation of high-frequency capillary waves. AB - The laser-induced surface deformation (LISD) technique was applied to generate high-frequency capillary waves on liquid surfaces up to several tens of kHz in a noncontact manner. The dynamic response of the fluid near the surface was theoretically derived under the condition of periodical radiation pressure. The result of the numerical calculation predicts the propagation of induced capillary waves out from the excitation region. The efficiency of the wave generation was experimentally examined by changing the width of the excitation laser beam at the surface. The observed LISD spectra were well reproduced by the theory, showing that the effective frequency band can be extended up to over 100 kHz. The propagation of the optically generated wave was measured with a laser probe sweeping the position of the observation. The spatial profile gives the surface tension and the shear viscosity of the sample liquid. The frequency domain measurement was also carried out and the spectrum obtained at a fixed point agrees with the theory, demonstrating the rapid measurement of frequency dependent phenomena. PMID- 16256662 TI - Improvement of interfacial adhesion and nondestructive damage evaluation for plasma-treated PBO and Kevlar fibers/epoxy composites using micromechanical techniques and surface wettability. AB - Comparison of interfacial properties and microfailure mechanisms of oxygen-plasma treated poly(p-phenylene-2,6-benzobisoxazole (PBO, Zylon) and poly(p-phenylene terephthalamide) (PPTA, Kevlar) fibers/epoxy composites were investigated using a micromechanical technique and nondestructive acoustic emission (AE). The interfacial shear strength (IFSS) and work of adhesion, Wa, of PBO or Kevlar fiber/epoxy composites increased with oxygen-plasma treatment, due to induced hydrogen and covalent bondings at their interface. Plasma-treated Kevlar fiber showed the maximum critical surface tension and polar term, whereas the untreated PBO fiber showed the minimum values. The work of adhesion and the polar term were proportional to the IFSS directly for both PBO and Kevlar fibers. The microfibril fracture pattern of two plasma-treated fibers appeared obviously. Unlike in slow cooling, in rapid cooling, case kink band and kicking in PBO fiber appeared, whereas buckling in the Kevlar fiber was observed mainly due to compressive and residual stresses. Based on the propagation of microfibril failure toward the core region, the number of AE events for plasma-treated PBO and Kevlar fibers increased significantly compared to the untreated case. The results of nondestructive AE were consistent with microfailure modes. PMID- 16256664 TI - Effect of salts on water viscosity in narrow membrane pores. AB - The effect of various salts on the viscosity, and by implication structure, of water in polymeric membrane pores of radius approximately 1.69 nm and low charge density has been studied. Permeation of pure water and various electrolyte solutions was analyzed using the Hagen-Poiseuille equation expressed in a ratio form to exclude membrane-specific quantities such as pore radius and length. The analysis produced viscosity ratios of electrolyte to pure water inside the membrane pores. Comparing the viscosity ratios inside the pores with their bulk counterparts showed that confinement significantly increased the sensitivity of water structure to the presence of ions. It has been found that, in relative terms in the pores, Cl- was a strong structure breaker, K+ was a moderately strong structure breaker, Na+ was a weak structure breaker, SO4(2-) was a weak structure maker, and Mg2+ was a strong structure maker. Predictive modeling of membrane separation performance would benefit from such effects being taken into account in cases where the pore ion concentrations may be high. PMID- 16256665 TI - Structure and thermal behavior of a layered silver hydroxyalkanecarboxylate. AB - The structure and thermal behavior of silver 16-hydroxyhexadecanoate (Ag-HHDA) have been investigated by using various analytical tools. The X-ray diffraction pattern was composed of a series of peaks that could be indexed to (0k0) reflections of a layered structure. Diffuse reflectance infrared Fourier transform spectroscopy revealed that the alkyl chains in Ag-HHDA as prepared were in an all-trans conformational state. Upon heating the sample, noticeable structural changes took place particularly in two temperature regions. The first structural change occurring at approximately 380 K was a partially irreversible one in which the binding state of carboxylate to silver converted from bridging into unidentate. A second dramatic structural change occurring at approximately 480 K was a totally irreversible process that could be associated with the decomposition of Ag-HHDA. All of these thermal characteristics of Ag-HHDA are comparable to those of silver stearate (Ag-STA). Separately, we have endeavored without success to intercalate polar molecules into the OH-group terminated layers in Ag-HHDA; the exfoliation of Ag-HHDA in various polar solvents was also unsuccessful. This is indicative of the presence of a rather stronger H-bond in Ag-HHDA, but the comparable thermal characteristics of Ag-HHDA and Ag-STA dictate that the thermal behavior of silver alkanoate is determined exclusively by the silver-to-carboxylate group interaction. This is in sharp contrast to the case of two-dimensional self-assembled monolayers for which the terminal functionalities play a crucial role in determining the structure and thermal stability of entire monolayers. PMID- 16256666 TI - Preparation of mesoporous carbon from commercial activated carbon with steam activation in the presence of cerium oxide. AB - Mesoporous carbon was prepared from the commercial activated carbon by steam activation with cerium oxide as catalyst. Steam activation with a catalyst loading of 0.5-2.0 wt% at 680-870 degrees C was examined. The surface area and pore size were evaluated by nitrogen adsorption at 77 K, and the structure of cerium oxide was characterized by XRD, XPS, and TEM. The results showed that the catalyst promoted the development of a mesopore at lower temperature (680-740 degrees C), and the mesopore was concentrated around 4-10 nm. The noncatalytic activation was advantageous in mesopore development and the catalyst would restrict the formation of mesopores at high temperature (800-870 degrees C). Higher loading of cerium oxide and higher activation temperature caused the aggregation of cerium oxide and then resulted in scattered pore size distribution. PMID- 16256667 TI - Molecular simulations of smectites intercalated with a zirconium complex ion. AB - Molecular mechanics simulations using the Cerius2 modeling environment have been applied to study the structures of dioctahedral smectites intercalated with the zirconium cation [Zr4(OH)12(H2O)12]4+. The substitutions in the silicate layers influence the sublimation energy in these types of intercalated layer structures. Charge distribution in the smectite layer was analyzed in relation to the interlayer structure. Tetrahedral substitutions in the smectite layer create preferential interlayer positions of the [Zr4(OH)12(H2O)12]4+ ion. A regular distribution of the tetrahedral substitutions in the smectite layer results in a better intercalant ordering in the interlayer space. Such a more homogeneous intercalant distribution results in a better interlayer pore size control. PMID- 16256668 TI - Spreading of aqueous SDS solutions over nitrocellulose membranes. AB - Experimental investigations were carried out on the spreading of small drops of aqueous SDS solutions over dry thin porous substrates (nitrocellulose membranes) in the case of partial wetting. The time evolution was monitored of the radii of both the drop base and the wetted area inside the porous substrate. The total duration of the spreading process was subdivided into three stages: the first stage: the drop base expands until the maximum value of the drop base is reached, the contact angle rapidly decreases during this stage; the second stage: the radius of the drop base remains constant and the contact angle decreases linearly with time; the third stage: the drop base shrinks and the contact angle remains constant. The wetted area inside the porous substrate expends during the whole spreading process. Appropriate scales were used with a plot of the dimensionless radii of the drop base, of the wetted area inside the porous substrate and the dynamic contact angle on the dimensionless time. Our experimental data show: the overall time of the spreading of drops of SDS solution over dry thin porous substrates decreases with the increase of surfactant concentration; the difference between advancing and hydrodynamic receding contact angles decreases with the surfactant concentration increase; the constancy of the contact angle during the third stage of spreading has nothing to do with the hysteresis of contact angle, but determined by the hydrodynamic reasons. It is shown using independent spreading experiments of the same drops on nonporous nitrocellulose substrate that the static receding contact angle is equal to zero, which supports our conclusion on the hydrodynamic nature of the hydrodynamic receding contact angle on porous substrates. PMID- 16256669 TI - A fluorescence emission study of the formation of induced premicelles in solutions of polyelectrolytes and ionic surfactants. AB - The interactions between PSS-co-BVE copolymers and ionic surfactants (anionic and cationic) in aqueous solution have been investigated using pyrene as a photophysical probe. Static and dynamic fluorescence determinations have been used to obtain information about the microenvironments formed between both species. Micropolarity studies using the I1/I3 ratio of the vibronic bands of pyrene and the behavior of the I(E)/I(M) ratio between the monomer and excimer emissions show the formation of hydrophobic domains. The interactions between the polyelectrolytes and the oppositely charged surfactants lead to the formation of induced premicelles at surfactant concentrations lower than the cmc of the surfactants. This aggregation process is assumed to be due to electrostatic attraction. At the same concentration, the excimer-to-monomer emission ratio shows its first peak. At higher surfactant concentrations, near the cmc, micelles with the same properties as those found in pure aqueous solution are formed. On the other side, systems containing an anionic surfactant do not show this behavior at low concentrations. There is no apparent dependence of the cac on the composition of the polymer, reinforcing the assumption that the electrostatic interactions induce the formation of the premicelles. The values of the cac's follow the same trend as for the cmc's, DTAC>DTAB>CTAC. The polarity of the induced premicelles, as measured by the I1/I3 ratio, also indicates that the microdomains formed by the longer chain surfactants are more hydrophobic than those of the shorter chain surfactants, as also happens with real micelles. PMID- 16256670 TI - Complexation of a hydrophobic thiazorylazophenol with Ni2+ at sodium dodecylsulfate micellar surface. AB - Complexation of thiazorylazododecylphenol (TADP) with Ni2+ at the surface of sodium dodecylsulfate (SDS) micelles has been spectrophotometrically studied. Complicated spectral changes are analyzed by a factor and multivariate analysis, which implies the formation of [TDAP(OH-)Ni]0 as well as a simple 1:1 complex of Ni2+ with TDAP at the micellar interface. All of the equilibrium constants required to describe this system are substantially affected by electrolyte concentrations. Coexistent electrolytes vary the surface potential of the micelle, and in turn influence the equilibria taking place on the micellar surface. The electrostatic potential estimated based on the equilibrium shifts is more negative than that calculated according to Poisson-Boltzmann theory, which simply involves electrostatic effects. This disagreement is possibly caused by different aqueous environments around the micelle from bulk solution, which also facilitate the formation of unusual complex such as [TDAP(OH-)Ni]0. PMID- 16256672 TI - Ostwald ripening of water-in-hydrocarbon emulsions. AB - The purposes of this study are to determine Ostwald ripening rates in water-in oil (W/O) emulsions and evaluate the potential application of the LSW theory to W/O emulsions. Water-in-oil emulsions were prepared by mixing water and hydrocarbon phases containing Span 83 and homogenizing. n-Heptane, n-decane, n dodecane, and n-tetradecane were used to obtain a range of solubilities. A linear increase in the cube of the droplet size with time was observed (within the initial period of 1-2 h after emulsion preparation), supporting the LSW theory. Based on this linear relationship, Ostwald ripening rates were determined to be 3.0 x 10(-24), 2.3 x 10(-24), 1.8 x 10(-24), and 5.8 x 10(-25) m3 s(-1) for water in-heptane, water-in-decane, water-in-dodecane, and water-in-tetradecane emulsions, respectively. These values are in agreement with theoretical predictions calculated using the LSW equation. It was observed that the ripening process gradually slowed, resulting in deviations from the LSW theory. This was attributed to the effect of the interfacial surfactant film through which the dispersed material has to diffuse during Ostwald ripening. This effect is not taken into consideration by the LSW theory. The results showed that Ostwald ripening of W/O emulsions was less sensitive to the nature of oil used and slower compared to O/W emulsions consisting of the same hydrocarbons. PMID- 16256671 TI - Monitoring of flocculation and creaming of sodium-caseinate-stabilized emulsions using diffusing-wave spectroscopy. AB - Diffusing-wave spectroscopy (DWS) has been used to study the stability of sodium caseinate-stabilized emulsions. The emulsions underwent creaming as a result of depletion flocculation when excess sodium caseinate was added. The creaming process was monitored over a 3-h period and each autocorrelation function was collected for 2 min to ensure adequate signal-to-noise ratio. The temporal variation of average particle size times the coefficient of viscosity of the continuous phase was derived from the backscattering measurements, and the variation of the scattering mean free path length with time was found from the backscattering and transmission measurements. It was confirmed that the creaming process was delayed at high oil concentrations, presumably due to the formation of oil droplet networks. PMID- 16256673 TI - Boundary effect on the drag force on a nonhomogeneous floc. AB - The boundary effect on the drag force acting on a spherical floc having a nonhomogeneous structure is examined by considering a spherical floc at the centerline of a cylindrical tube. The floc is simulated by an entity having a two layer structure, and its porous nature mimicked by varying the relative magnitude of the permeability of its inner and outer layers. The results of numerical simulation reveal that the tube wall has the effect of compressing the streamlines and vorticity contours. Also, as in the case of a rigid entity, the wake in the rear region of a floc, which arises from the convective motion of the fluid, is depressed. For fixed volume-averaged permeability, the influence of the tube wall on the behavior of a heterogeneous floc is more significant than that on the behavior of a homogeneous floc, and the influence varies with the structure of the former. The heterogeneous structure of a floc leads to a deviation in the modified drag coefficient-Reynolds number relation from a Stokes law-like correlation. The smaller the average permeability of a floc the greater the deviation, but the presence of the tube wall has the effect of reducing the deviation. PMID- 16256674 TI - Effect of sodium chloride on association behavior of poly(ethylene oxide) poly(propylene oxide)-poly(ethylene oxide) block copolymer in aqueous solutions. AB - The effect of sodium chloride (NaCl) upon the thermally induced association behavior of poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) (PEO PPO-PEO) block copolymer, Pluronic P103, has been investigated using pyrene fluorescence spectroscopy. The critical micellization temperature (CMT) of Pluronic P103 in aqueous solution is decreased by the addition of NaCl. The standard enthalpy and entropy of micellization for Pluronic P103 in water are increased in the presence of small amounts of NaCl, but further addition of NaCl decreases the standard enthalpy and entropy of micellization. The I1/I3 ratio of pyrene in aqueous Pluronic P103 solutions at temperature below the CMT decreases with increases of NaCl concentration, which is related to the decrease of PPO solubility. The decrease in polarity of the PPO shifts the CMT toward lower temperature. PMID- 16256675 TI - Interpretation of structure formation during the sol-gel transition of a resorcinol-formaldehyde solution by population balance. AB - Growth of colloidal particles formed during the sol-gel transition of a resorcinol-formaldehyde (RF) solution was simulated based on the population balance equation by using the discrete-sectional model (DSM). During the early stage of the sol-gel transition, the transient change of sizes of colloidal particles estimated by this method agreed well with the previous experimental observation by dynamic light scattering (DLS), which confirmed the influence of the catalyst concentration of a starting RF solution on the growth rate of the particles. From the size distribution of colloidal particles predicted at the gelation time, the surface area of a RF hydrogel after the completion of the sol gel transition was estimated, which coincided with the BET surface area of a RF aerogel because the porous structure of a hydrogel was maintained and few micropores were formed during supercritical drying. PMID- 16256676 TI - NMR diffusion studies of translational properties of oil inside core-shell latex particles. AB - The diffusion behavior of core-shell latex particles with a liquid core of hexadecane and a solid polystyrene shell in water solution has been studied using the pulsed field gradient spin-echo (PFG-SE) NMR technique. The apparent diffusion coefficient and the root mean square displacement of oil were strongly dependent on the diffusion time Delta. With increasing diffusion time, the obstructing effect from the particle wall caused a decrease in the apparent oil diffusion coefficient. The root mean square displacement of oil inside the particle core was constant for all diffusion times and was used for the calculation of the particle radius. The volume fraction dependence of the apparent diffusion coefficient was found to be roughly consistent with the hard sphere model. The diffraction pattern in the echo decay predicted from the q space formalism for molecules diffusing inside a spherical cavity was almost completely smeared out due to polydispersity and wall relaxation effects. It was observed that 10-20% (w/w) of the particle shell consisted of hexadecane. This fact imparted a slow component to the echo decay, since the exchange time between oil in the shell and oil in the cavity was slow, which further contributed to the smearing out of the diffraction pattern. It was concluded that by using the core shell concept very good signal-to-noise is obtained in the PFG-SE experiment, thus making possible studies of translational properties of colloidal particles in different environments to an extent that previously has been very difficult to perform. PMID- 16256677 TI - Thickness and refractive index measurements using multiple beam interference fringes (FECO). AB - We report on the use of optical interferometry employing fringes of equal chromatic order (FECO) in a surface force apparatus (SFA) to determine film thicknesses and refractive indices of confined media for a wide range of separations. In particular, we show how to calculate the surface separation (film thickness) based on two fringes whose contact position was not measured. We discuss the measurement accuracy, and though the theoretical accuracy is 1 A for all separations, we show that in practice, for large separations, it is very hard to get to this accuracy. PMID- 16256678 TI - Rheology of a lyotropic mesophase through a stress-relaxation experiment. AB - We report on the first rheological study of the structural relaxations in a nematic liquid crystalline phase. Linear dynamic and transient shear experiments were applied to a polydomain nematic phase of the CTAB/water system: a liquid crystalline mixture composed of 28 wt% CTAB at 35 degrees C. The decay of the shear modulus, G(t), was analyzed using the CONTIN inverse Laplace transform to obtain a distribution of relaxation times which were compared with ones from the usual fitting procedure based on a generalized Maxwell model. The behavior of the nematic lyotropic structure of the CTAB/water system is characterized by the presence of both slow and fast relaxation times. These were interpreted as being due to a progressive loss of the lyotropic domain orientation and to the breaking/reforming process of the cylindrical aggregates, respectively. PMID- 16256679 TI - The use of size distributions of circular domains in Langmuir monolayers for determining physical parameters of surfactants. AB - An analysis of equilibrium size distributions of circular liquid-condensed (LC) domains in amphiphilic monolayers at the air/water interface, gleaned from microscopic images, is presented in terms of Gaussians fitted to a theoretical expression derived previously. It is demonstrated how, in principle, important properties of the monolayer, such as molecular dipole moments of surfactant head groups and line tension of the liquid-condensed/liquid-expanded phase boundary, can be obtained by combining the results of this simplified analysis with information from X-ray diffraction or surface pressure measurements. PMID- 16256680 TI - Preparation and characterization of nanoaggregates of poly(ethylene oxide)-b polymethacrylate and poly-L-lysine. AB - Poly(ethylene oxide)-b-polymethacrylate (PEO-b-PMA), one of the double hydrophilic block copolymers, has proved to the form nanoaggregates with poly-L lysine (PLS). This was confirmed by turbidimetry, zeta-potential measurements, and dynamic light scattering. The nanoaggregate formation is induced by electrostatic charge neutralization of the PMA block with PLS. The properties of the aggregates are affected by PLS concentration as well PEO-b-PMA concentration. The aggregates have potential applications in biomedical science. PMID- 16256681 TI - Influence of growth phase on bacterial cell electrokinetic characteristics examined by soft particle electrophoresis theory. AB - The influence of incubation time on the electrokinetic properties of Escherichia coli, Pseudomonas putida, Alcaligenes faecalis, and Alcaligenes sp., was examined by electrophoretic mobility measurements and the results were discussed based on Ohshima's soft particle theory. The electrophoretic mobility of E. coli plotted against incubation time revealed that the mobility gradually increased from the outset of incubation to 7 h, which corresponded to the exponential growth and early stationary phase, and then decreased. For all strains, electrophoretic mobility leveled off to nonzero values in accordance with the increase in ionic concentration, which was a characteristic feature of soft particles. Soft particle analysis was carried out at the outset of incubation and the time when mobility reached a maximum in order to obtain the spatial charge density (ZN) and cell surface softness (1/lambda). ZN at the maximum electrophoretic mobility was more negative than that at the outset of incubation for E. coli, P. putida, and A. faecalis, while the ZN of Alcaligenes sp. almost remained unchanged. 1/lambda decreased for E. coli, but increased for Alcaligenes sp. and A. faecalis. These findings indicated that the cell growth phase affects both ZN and 1/lambda, although the dependencies of these parameters are unique to each bacterial strain. PMID- 16256682 TI - Postembryonic changes in the response properties of wind-sensitive giant interneurons in cricket. AB - The intensity-response (I-R) relations for four wind-sensitive giant interneurons (GIs 8-1, 9-1, 9-2 and 9-3) in the fourth-, sixth- and last-instar nymphs of the cricket, Gryllus bimaculatus, were investigated using a unidirectional air current stimulus in order to explore the functional changes of GIs during postembryonic development. Contrary to our expectations, the response properties of GIs in nymphs were largely different from those in adults. The response magnitude of GI 8-1 in an intact cricket decreased during development, i.e. the GI in younger insects showed a larger response magnitude. Although the response magnitudes of GIs 9-1 and 9-2 were almost identical during the nymphal period, a significant decrease was observed after the imaginal ecdysis. During the nymphal period, the response magnitude of GI 9-3 increased according to the developmental stage. However, it decreased significantly after the imaginal ecdysis. We also investigated the response magnitudes of the GIs in nymphs after unilateral cercal ablation. From the results of ablation experiments, the changes in excitatory and/or inhibitory connections between filiform hairs and each GI during postembryonic development were revealed. PMID- 16256683 TI - Longevity cost of reproduction for males but no longevity cost of mating or courtship for females in the male-dimorphic dung beetle Onthophagus binodis. AB - Life history theory predicts a trade-off between current and future reproduction. Despite a wealth of research on the cost of reproduction for females, there have been very few studies that have looked at the cost of reproduction for males. Longevity is closely related to the opportunity for future reproduction, and thus decreased longevity in response to current reproductive effort has been used as a measure of the cost of reproduction. Here we examine the cost of reproduction for males and females in the dung beetle Onthophagus binodis. Like many onthophagines, O. binodis exhibit dimorphic male morphology; major males develop a large pronotal horn while minor males remain hornless. Alternative morphologies are associated with alternative reproductive tactics. Thus, we ask whether major and minor males pay different costs of reproduction. We found that in contrast to previous work on Diptera, mating is not costly in terms of reduced longevity for female dung beetles. Despite a longevity cost of reproduction for males, we found no evidence for differential longevity costs associated with alternative reproductive tactics. PMID- 16256684 TI - Costs of mating and egg production in female Callosobruchus chinensis. AB - Costs of reproduction include the costs of mating and egg production. Specific techniques such as irradiation or genetic mutation have been used to divide the expense into costs of mating and egg production in previous studies. We tried to divide the costs in the adzuki bean beetle, Callosobruchus chinensis (Coleoptera: Bruchidae), which needs some kinds of bean as an oviposition substrate. Mated females that were not allowed to lay eggs had a shorter life span than virgin females, but they had a longer life span than mated females that were allowed to lay eggs. The results showed two independent significant costs, mating and egg production, on the life span in C. chinensis. Costs of mating, however, include the costs of sexual harassment by males and copulation itself, and we need further studies to divide the costs. The present method for dividing the cost of reproduction into costs of mating and egg production can be applied to a broad taxonomic range of insect species, and thus it will be a useful model system for inter-specific comparisons of costs of mating and egg production. PMID- 16256685 TI - Suppression of the prophenoloxidase system in Rhodnius prolixus orally infected with Trypanosoma rangeli. AB - Investigations were carried out to compare aspects of the prophenoloxidase (proPO)-activating pathway in Rhodnius prolixus hemolymph in response to oral infection and inoculation of the insects with two developmental forms of Trypanosoma rangeli epimastigotes strain H14. In vivo experiments demonstrated that in control insects fed on uninfected blood, inoculation challenge with short epimastigotes resulted in high phenoloxidase (PO) activity. In contrast, previous feeding on blood containing either short or long epimastigotes was able to suppress the proPO activation induced by thoracic inoculation of the short forms. In vitro assays in the presence of short epimastigotes demonstrated that control hemolymph or hemolymph provided by insects previously fed on blood containing epimastigotes incubated with fat body homogenates from control insects significantly increased the PO activity. However, fat body homogenates from insects previously fed on blood containing epimastigotes, incubated with hemolymph taken from insects fed on control blood or blood infected with epimastigotes, drastically reduced the proPO activation. The proteolytic activity in the fat body homogenates of control insects was significantly higher than in those obtained from fat body extracts of insects previously fed on blood containing epimastigotes. These findings indicate that the reduction of the proteolytic activities in the fat body from insects fed on infected blood no longer allows a significant response of the proPO system against parasite challenge. It also provides a better understanding of T. rangeli infection in the vector and offer novel insights into basic immune processes in their invertebrate hosts. PMID- 16256686 TI - Free amino acids are important for the retention of protein and non-protein meals by the midgut of Aedes aegypti females. AB - There is a relationship between the normal progress of digestion and the retention or elimination of the proteins ingested with the meal by Aedes aegyti females. The addition of soybean trypsin inhibitor (STI) to a protein meal prevented digestion and resulted in a rapid elimination of the undigested proteins. The addition of a mix of free amino acids to a protein meal together with STI resulted in a significant increase in the retention of the undigested proteins during the first 10-15 hrs after feeding. The effect of the free amino acids on the retention of the proteins was concentration-dependent between 250 microg/ml and 5 mg/ml. Free amino acids were also important for the retention of non-protein meals. When females were fed a meal containing FITC-dextran (20 kD), most of this compound was eliminated into the feces by 10 hrs; the addition of free amino acid resulted in a significant increase in the retention of the FITC dextran by the midgut during the first 15 hrs after feeding. The presence of free amino acids in the midgut lumen seems to be an important signal used by the mosquito to regulate the retention of the meal. PMID- 16256687 TI - Increased EAG responses of tortricid moths after prolonged exposure to plant volatiles: evidence for octopamine-mediated sensitization. AB - As measured by electroantennograms (EAG), both male and female obliquebanded leafrollers, Choristoneura rosaceana (Harris), and redbanded leafrollers, Argyrotaeniavelutinana (Walker), were similarly sensitive to host-related plant volatiles: trans-2-hexenal, benzaldehyde, 1-hexenol, cis-3-hexen-1-ol, geraniol, linalool, (+)-limonene, hexenal and trans-2-hexenol. Females of both species were similarly sensitive to the shared major component of their sex-attractant pheromone ((Z)11-14:Ac). Continuous 60 min pre-exposure of male and female C. rosaceana and A. velutinana to successively higher concentrations of a mixture of the nine plant volatiles in Teflon chambers with continuous air exchange caused a dosage-dependent increase in subsequent responsiveness (sensitization) to green leaf volatiles, as measured by EAGs. In addition, 60 min of pre-exposure of male C. rosaceana to certain individual volatiles ((+)-limonene, geraniol, benzaldehyde) increased EAGs nearly as much as did the mixture of nine volatiles. Pre-exposures to the nine plant-volatile mixture at concentrations achieved by 100 microg and 1 mg loading dosages in 100 microl of mineral oil significantly increased EAG depolarization to pheromone (cross-sensitization) in males but not females of both moth species. Antennae of male C. rosaceana pre-injected with 100 microg of octopamine (OA) without volatile pre-exposure exhibited sensitization nearly identical to that induced by pre-exposing moths to sensitizing concentrations of the plant-volatile mixture. Moreover, injection of the OA antagonist chlorpromazine (CP) blocked sensitization by the plant-volatile pre exposure. Collectively, these findings suggest that exposures of tortricid moths to certain host-plant related volatiles may modulate subsequent olfactory sensitivity to behaviorally relevant chemical cues and that plant-volatile induced sensitization may be octopamine mediated. PMID- 16256688 TI - Structure and function of the antennal sensilla of the palm weevil Rhynchophorus palmarum (Coleoptera, Curculionidae). AB - The distribution, fine structure and function of the sensilla present on the antennal club of Rhynchophorus palmarum were studied. No sex dimorphism was observed. Scanning and transmission electron microscopy showed five types of hair like structures, four of which were evenly distributed on the antennal club. Two types of hair (IV and V) showed wall pores, a characteristic of olfactory sensilla. The antenna numbers 11,190 +/- 3040 type IV and 7360 +/- 1500 type V hairs. Using single sensillum recording, we identified 17 types of olfactory receptor neurons (ORNs) on the basis of their responses to pheromone and host plant odors, triggering synergic behavioral responses. We characterized highly specific and sensitive ORNs tuned to the aggregation pheromone (18% ORNs; 0.01-1 ng response threshold) and to host plant odors such as ethyl acetate, ethanol, acetoin and guaiacol (10% ORNs; 1-10 ng response threshold). Eleven percent of the ORNs were more generalist, responding to several odors with low sensitivity. Nine percent of the ORNs showed a complex pattern of responses, being co activated by the pheromone and plant odors. This suggests an interaction at the sensory neuron level between pheromone and plant odors, triggering synergic behavioral responses. PMID- 16256689 TI - Cowpea (Vigna unguiculata) vicilins bind to the peritrophic membrane of larval sugarcane stalk borer (Diatraea saccharalis). AB - In this work, we show that vicilins from two Vigna unguiculata (cowpea) genotypes, Epace-10 and IT 81D-1045, which are susceptible and resistant to attack by the cowpea weevil Callosobruchus maculatus, respectively, associate with the peritrophic membrane (PM) from larvae of Diatraea saccharalis. Solutions with increasing concentrations of vicilins were incubated with PM of the larvae and subsequently analysed by electrophoresis with SDS. It was observed that the majority of the bands of approximately 50,000 Da (characteristic of vicilins) did not appear in the separating gel and only lower molecular weight polypeptides were seen. When vicilins were incubated with PM, and the solution was then heated after the incubation, the band pattern in the gel appeared completely different. It was observed that the vicilins were being hydrolysed by proteinases associated with the PM. When the incubated samples were heated after the reaction, the major bands reappeared, demonstrating that most of the vicilin molecules had bound to the PM of D. saccharalis. These results suggest that when the vicilins are in contact with the PM they are bound and also digested by the PM of this insect. The major and several minor proteinases from the PM were extracted with Triton X 100 and their activity and the inhibition of this activity were analysed by ingel assays. Based on the effects of proteinase inhibitors, the PM-associated activity is due to serine class proteinases. Larvae of D. saccharalis were fed on artificial diets containing purified vicilins from Epace-10 or IT 81D-1045 seeds. Vicilins from Epace-10 did not affect the larval development, while IT 81D-1045 vicilins reduced significantly the survival rate of the sugar cane borer. PMID- 16256690 TI - Oxygen consumption and body temperature of active and resting honeybees. AB - We measured the energy turnover (oxygen consumption) of honeybees (Apis mellifera carnica), which were free to move within Warburg vessels. Oxygen consumption of active bees varied widely depending on ambient temperature and level of activity, but did not differ between foragers (>18 d) and middle-aged hive bees (7-10 d). In highly active bees, which were in an endothermic state ready for flight, it decreased almost linearly, from a maximum of 131.4 microl O(2) min(-1) at 15 degrees C ambient temperature to 81.1 microl min(-1) at 25 degrees C, and reached a minimum of 29.9 microl min(-1) at 40 degrees C. In bees with low activity, it decreased from 89.3 microl O(2) min(-1) at 15 degrees C to 47.9 microl min(-1) at 25 degrees C and 14.7 microl min(-1) at 40 degrees C. Thermographic measurements of body temperature showed that with increasing activity, the bees invested more energy to regulate the thorax temperature at increasingly higher levels (38.8 41.2 degrees C in highly active bees) and were more accurate. Resting metabolism was determined in young bees of 1-7 h age, which are not yet capable of endothermic heat production with their flight muscles. Their energy turnover increased from 0.21 microl O(2) min(-1) at 10 degrees C to 0.38 microl min(-1) at 15 degrees C, 1.12 microl min(-1) at 25 degrees C, and 3.03 microl min(-1) at 40 degrees C. At 15, 25 and 40 degrees C, this was 343, 73 and 10 times below the values of the highly active bees, respectively. The Q(10) value of the resting bees, however, was not constant but varied in a U-shaped manner with ambient temperature. It decreased from 4.24 in the temperature range 11-21 degrees C to 1.35 in the range 21-31 degrees C, and increased again to 2.49 in the range 30-40 degrees C. We conclude that attempts to describe the temperature dependence of the resting metabolism of honeybees by Q(10) values can lead to considerable errors if the measurements are performed at only two temperatures. An acceptable approximation can be derived by calculation of an interpolated Q(10) according to the exponential function (V(O(2))=0.151 x 1.0784(T(a))) (interpolated Q(10)=2.12). PMID- 16256691 TI - Effect of the terminal amino group of a linker arm and its length at the C5 position of a pyrimidine nucleoside on the thermal stability of DNA duplexes. AB - 2'-Deoxyuridine derivatives bearing a substituent at the C5-position, which has a different chain length and a different functional group (methyl or amino), were synthesized and incorporated into oligodeoxyribonucleotides. The effect of the substituent groups in the major groove on the stability of the duplexes was investigated by UV melting experiments. It was found that the stabilization of these duplexes by a terminal amino group depended on the length of a linker arm. PMID- 16256692 TI - Synthesis of a dA-dT base pair analogue and its effects on DNA-ligand binding. AB - Two nucleoside derivatives containing the base analogues 3-deazaadenine and 3 methyl-2-pyridone have been prepared as analogues of dA and dT, respectively. After conversion into the appropriately protected phosphoramidites, DNA sequences were prepared with site-specifically placed analogues. When present in a duplex DNA sequence, the analogues result in the deletion of one or both of the hydrogen bonding functional groups (the N3-nitrogen of dA and the O2-carbonyl of dT) present in the minor groove. Binding by two ligands, 4',6-diamidine-2-phenyl indole (DAPI) and Hoechst 33258 in the minor groove has been probed using a variety of DNA sequences. These sequences contain a d(GAATTC)2 core with analogue nucleosides substituted for one or more of the dA and dT residues. DAPI bound strongly to any sequence that contained both O2-carbonyls of the central two dT residues. The presence of a dc3A residue did in some cases enhance binding. With one of the central O2-carbonyls deleted, the binding was noticeably reduced, and with both absent, no significant binding could be detected. Similar although less dramatic results were observed with Hoechst 33258 binding to analogue sequences. PMID- 16256693 TI - Inhibition and structure-activity studies of methionine hydroxamic acid derivatives with bacterial peptide deformylase. AB - The posttranslational deformylation of N-formyl-Met-polypeptides by the metalloenzyme, peptide deformylase, is essential for bacterial growth. Methionine hydroxamic acid derivatives were found to inhibit recombinant Escherichia coli peptide deformylase activity containing either zinc or cobalt. The binding of methionine hydroxamate and hydrazide inhibitors to cobalt-substituted deformylase caused spectral changes consistent with the formation of a pentacoordinate metal complex similar to that of actinonin, a psuedopeptide hydroxamate inhibitor. The spectral and kinetic data support the binding of these N-substituted L-methionine derivatives in a reverse orientation with respect to N-formyl-Met-peptide substrates within the active site. Based on this hypothesis a second generation of N-substituted methionyl hydroxamic acids were evaluated and found to possess greater inhibitory potency. These results may provide the basis for the design of more potent and selective deformylase inhibitors as potential antibacterial agents. PMID- 16256694 TI - Photo-cross-linked oligonucleotide duplex as a decoy-DNA for inhibition of restriction endonuclease activity. AB - As a novel type of regulator molecule for DNA-recognizing proteins, a photo-cross linked oligonucleotide duplex was designed and synthesized. The molecule regulated the activity of a restriction endonuclease by being recognized as a substrate. This type of regulating molecule is regarded as a decoy-DNA. 4,5',8-[4 Aminoethylaminomethyl]-trioxalen (aeAMT) was conjugated with an oligodeoxyribonucleotide (ODN) at the 5'-end and the aeAMT was cross-linked with the thymine residue of the complementary oligonucleotide upon UVA irradiation. The terminally cross-linked oligonucleotides, singly clipped (SC) decoy-DNA, acquired thermal stability. An oligonucleoside phosphorothioate (OPT) was also introduced as one or both components, yielding three types of decoy-DNAs, SC-ODN ODN (SC.DD), SC-OPT-ODN (SC.SD), and SC-OPT-OPT (SC.SS). The SC decoy-DNAs inhibited the function of the restriction endonuclease, AatII, in a sequence specific and concentration-dependent manner with an appreciable IC50 value (1.3 microM for SC.DD, 0.016 microM for SC.SD, 0.002 microM for SC.SS). The SC decoy DNAs were found to be effective for regulating the DNA recognizing proteins. PMID- 16256695 TI - PLP and PMP radicals: a new paradigm in coenzyme B6 chemistry. AB - Enzymes frequently rely on a broad repertoire of cofactors to perform chemically challenging transformations. The B6 coenzymes, composed of pyridoxal 5'-phosphate (PLP) and pyridoxamine 5'-phosphate (PMP), are used by many transaminases, racemases, decarboxylases, and enzymes catalyzing alpha,beta and beta,gamma eliminations. Despite the variety of reactions catalyzed by B6-dependent enzymes, the mechanism of almost all such enzymes is based on their ability to stabilize high-energy anionic intermediates in their reaction pathways by the pyridinium moiety of PLP/PMP. However, there are two notable exceptions to this model, which are discussed in this article. The first enzyme, lysine 2,3-aminomutase, is a PLP dependent enzyme that catalyzes the interconversion of L-lysine to L-beta-lysine using a one-electron-based mechanism utilizing a [4Fe-4S] cluster and S adenosylmethionine. The second enzyme, CDP-6-deoxy-L-threo-D-glycero-4-hexulose-3 dehydrase, is a PMP-dependent enzyme involved in the formation of 3,6 dideoxysugars in bacteria. This enzyme also contains an iron-sulfur cluster and uses a one-electron based mechanism to catalyze removal of a C-3 hydroxy group from a 4-hexulose. In both cases, the participation of free radicals in the reaction pathway has been established, placing these two B6-dependent enzymes in an exclusive class by themselves. PMID- 16256696 TI - The development of photometric sensors for boronic acids. AB - Boronic acids bind certain 1,2- and 1,3-diols with high affinity through reversible formation of boronate esters. They have been utilized as the recognition moiety for artificial receptors, particularly receptors for carbohydrates that have cis-diol moieties. Therefore, sensors for boronic acids could serve as universal reporters for monitoring boronate formation. This paper reports the design and synthesis of a series of photometric chemosensors for phenylboronic acid using diethanolamine as the recognition moiety. Diethanolamine, which binds strongly to boronic acids, has been linked to three different types of optical reporters. A photoinduced electron transfer system based on the anthracene fluorophore has been used to create sensors that show up to a fivefold increase in fluorescent intensity in the presence of millimolar concentrations of phenylboronic acid. Sensor designs based on the restriction of free rotation of extended pi systems and on the perturbed electronic properties of azo dyes are also included. This work demonstrates that sensors based on several different designs can be used for the detection of boronic acids. PMID- 16256697 TI - Inverse acyl phosph(on)ates: substrates or inhibitors of beta-lactam-recognizing enzymes? AB - Acyl phosph(on)ates represent a new class of inhibitors of beta-lactam recognizing enzymes. Previously described members of this class were aroyl phosph(on)ates. These compounds have been shown to acylate and/or phosphylate the active site serine residue, leading to either transient or essentially irreversible inhibition [Li, N., and Pratt, R. F. (1998) J. Am. Chem. Soc.120, 4264-4268]. The present paper describes the synthesis and evaluation as inhibitors of an inverse pair of acyl phosph(on)ates that incorporate the amido side chain that represents a major substrate specificity determinant of these enzymes. Thus, N-(phenylacetyl)glycyl phenyl phosphate and benzoyl N (benzyloxycarbonyl)aminomethyl phosphonate were prepared. The former of these compounds was found to be a substrate of typical class A and C beta-lactamases and of the DD-peptidase of Streptomyces R61; it thus acylates the active site serine. In contrast, the latter compound was an irreversible inhibitor of the above enzymes, probably by phosphonylation of the active site serine. With each of these enzymes therefore, the amido side chain rather than the acyl group dictates the orientation of the bound phosph(on)ate and thus the mode of reaction. PMID- 16256698 TI - Cyclic analogs of insect oostatic peptides: synthesis, biological activity, and NMR study. AB - Cyclic peptides 2a-2c, derived from the sequence of the C-terminal shortened analogs of the oostatic decapeptide H-Tyr-Asp-Pro-Ala-Pro-Pro-Pro-Pro-Pro-Pro-OH (1a), were synthesized and assayed on their effect in a reproduction of the flesh fly Neobellieria bullata. The cyclization of the N-terminal linear tetra- and pentapeptides 1b and 1c to the cyclotetra- and cyclopentapeptides 2b and 2c decreased the oostatic activity by one order of magnitude. The cyclodecapeptide 2a, which emerged spontaneously during the pentapeptide cyclization, was quite inactive. Comparative 1H and 13C NMR study on a conformation of the cyclopeptides 2a-2c, and their linear precursors 1b and 1c revealed that a space structure of the cyclic analogues 2b and 2c is too restricted to adopt a biological conformation necessary for receptor binding and therefore only minor oostatic activity is observed after their application. The lack of the oostatic activity in the case of the more flexible dimeric analogue 2a is ascribed to the size of its molecule and its overall shape that is not compatible with a receptor binding. PMID- 16256699 TI - Hexanoate synthase, a specialized type I fatty acid synthase in aflatoxin B1 biosynthesis. AB - In fungi, fatty acids are biosynthesized by large multifunctional enzyme complexes, the fatty acid synthases (FASs), which catalyze chain assembly in an iterative manner. Many fungal secondary metabolites contain fatty acid moieties, and it is often unclear whether they are recruited from primary metabolism or are biosynthesized de novo by secondary metabolic FASs. The most convincing evidence of such a dedicated FAS comes from the biosyntheses of aflatoxin (AF) and sterigmatocystin (ST) in certain species of the filamentous fungus Aspergillus. Incorporation studies in AF and genetic analyses of ST and AF biosynthesis strongly suggest that their biosyntheses begin with the production of a C6 fatty acid by a specialized FAS. The genes encoding the alpha (hexA) and beta (hexB) subunits of this hexanoate synthase (HexS) from the AF pathway in Aspergillus parsiticus SU-1 were cloned and both their gDNAs and cDNAs were sequenced and their transcriptional ends analyzed. Translated amino acid sequences are predicted to result in proteins of 181.3 and 210.5 kDa, for HexA and HexB, respectively. Comparison of the HexA and HexB sequences with those of the ST FAS subunits and primary metabolic FASs indicated that the secondary metabolic enzymes are members of a well-defined subclass of the FAS family. Phylogenetic predictions and an analysis of GC-bias in AF and ST pathway genes compared with primary metabolic Aspergillus genes were used as a basis to propose a route for the evolution of the AF and ST clusters. PMID- 16256700 TI - Building fluorescent sensors for carbohydrates using template-directed polymerizations. AB - The ability to custom-make fluorescent sensors for different analytes could have a tremendous impact in a variety of areas. Template-directed polymerization or molecular imprinting seems to be a promising approach for the preparation of high affinity and specific binding sites for different template molecules. However, the application of molecular imprinting in the preparation of fluorescent sensors has been hampered by the lack of suitable fluorescent tags, which would respond to the binding event with significant fluorescence intensity changes. We have designed and synthesized a fluorescent monomer (1) that allows for the preparation of fluorescent sensors of cis diols using molecular imprinting methods. This monomer has been used for the preparation of imprinted polymers as sensitive fluorescent sensors for D-fructose. The imprinted polymers prepared showed significant fluorescence intensity enhancement upon binding with the template carbohydrate. PMID- 16256701 TI - Regioselective oxidation of steroids by a manganese porphyrin carrying metal coordinating groups. AB - A manganese porphyrin having four 2,2'-bipyridyl groups on its meso positions was synthesized. In the presence of Cu2+ ions it catalyzes the regioselective oxidation of steroid substrates carrying auxiliary metal coordinating groups. PMID- 16256702 TI - Intranasal absorption of granulocyte-colony stimulating factor (G-CSF) from powder formulations, in sheep. AB - Granulocyte-colony stimulating factor (G-CSF) was administered to sheep in three different nasal formulations and as a subcutaneous injection. The nasal formulations were: a solution containing L-alpha-lysophosphatidylglycerol (LPG), a powder formulation comprising small starch microspheres (SSMS) and a powder formulation comprising SSMS and LPG. Absorption of G-CSF was assessed directly by quantitation in plasma and indirectly by measurement of the pharmacodynamic response in terms of leucocyte and neutrophil counts. After the nasal delivery of the G-CSF powder formulation containing SSMS and LPG the absorption of G-CSF was significantly higher (P<0.01) than that from the simple nasal solution or the powder without the enhancer, but the resulting pharmacological response was not significantly different. The bioavailability of G-CSF from the powder formulation containing SSMS and LPG relative to the subcutaneous injection was 8.4% (+/-3.4). We also found that at the respective G-CSF doses investigated, the pharmacodynamic response of this nasal formulation, was similar to that obtained after the subcutaneous administration. The study indicates that the powder formulation containing enhancers could offer an alternative delivery route for G CSF in the form of intranasal administration. PMID- 16256704 TI - Structure-activity relationships of some indolo[3,2-c]quinolines with antimalarial activity. AB - The synthesis, physicochemical characterization and in vitro antimalarial activity of a series of indolo[3,2-c]quinolines (9a-f) are described. There is only a poor correlation between the activity and hydrophobicity. In contrast, 33% of the observed variation in antimalarial activity can be attributed to the size of the side chain attached to position 9 of the indoloquinoline ring. An increase in the size of this dibasic side chain generally results in a reduction in activity, suggesting that it is accommodated in a site/cavity of limited size on the receptor. More significantly, the charge on the distal nitrogen (N3) on the side chain, located 10-11 A from the quinoline N, could account for 75% of the observed variation. Since a large charge on N3 is associated with improved antimalarial activity, it is suggested that N3 is protonated and functions as a H bond donor in the drug-receptor interaction. PMID- 16256703 TI - Route dependent pulmonary first-pass metabolism of a series of biphenylacetic acid esters in rats. AB - The influence of route of administration of a series of biphenylacetic acid aliphatic esters on their pulmonary first-pass metabolism in rats was investigated. The results were compared with in vitro data and with the physicochemical properties of the esters. Pre- and post-absorptive first-pass metabolism was assessed by comparing area under blood concentration-time curves after intra-arterial administration of the esters with those seen after intravenous (to assess post-absorptive) and intratracheal (to assess pre absorptive) administration. The extent of pulmonary metabolism was dependent on the route of administration. For all the esters studied there was insignificant post-absorptive first-pass extraction, however there was a large pre-absorptive first-pass extraction (greater than 80%). Enzyme kinetics derived in vitro using the 'well-stirred' model (Dickinson and Taylor, 1995. Int. J. Pharm. 116, 231 236) predicted the extent of post-absorptive but not the pre-absorptive, first pass metabolism. PMID- 16256705 TI - Gastric emptying in patients with insulin dependent diabetes mellitus and bioavailability of thioctic acid-enantiomers. AB - The objective of this study was to determine the impact of prolonged gastric emptying in patients with insulin dependent diabetes mellitus (IDDM) on the bioavailability of the R(+)- and S(-)-thioctic acid (TA) enantiomers. Gastric emptying time (GET) was assessed in 30 healthy volunteers and 22 patients with IDDM using sequential ultrasonography after a standardized solid-liquid test meal. Pharmacokinetics and absolute bioavailability (F) of the TA-enantiomers were studied using a randomized, open two-way crossover design with administrations of oral and intravenous single doses of 200 mg rac-TA. GET in healthy subjects was 134.7+/-21.6 min, the normal range was calculated from 88.3 to 181.1 min. The mean GET in all IDDM patients was significantly prolonged (178.2+/-28.1 min; P<0.001). Only 50% of the patients (n=11) were found to have normal GET (group A), the other half of the population (n=11) were considered to have delayed GET (group B). Mean GET values were 156.9+/-21.5 in group A (P=0.028) and 199.4+/-13.9 min in group B, respectively, suggesting that gastric motility is significantly different from non-diabetic controls even in patients with apparently normal gastric emptying. Times to peak plasma concentrations (t(max)) of both TA-enantiomers were similar in both groups and thus, unrelated to measures of gastric emptying. In contrast, maximum concentrations (C(max)) and area-under-the-curve values (AUC) of both enantiomers were reduced by about 30% in patients with delayed GET. Although these differences resulted in statistical significance for the AUC of both enantiomers (P<0.05), linear regression analysis showed only modest correlation between GET and the extent of TA-enantiomer absorption (r2=0.31 and 0.22 for R(+)-/S(-)-TA, respectively). The study suggests that prolonged gastric emptying is frequently present in IDDM. Delayed gastric emptying, however, does not substantially affect the rate and extent of absorption of both TA-enantiomers. PMID- 16256706 TI - Evaluation of LC methods for the separation of phenoxymethylpenicillin and its related substances. AB - Five isocratic reversed-phase liquid chromatography (LC) methods have been examined for the separation of phenoxymethylpenicillin and its related substances. A method previously selected for the analysis of benzylpenicillin gave the best selectivity. The mobile phase of this method is composed of 0.5 M phosphate buffer (pH 3.5)-water-methanol (10:50:40), the amount of organic modifier was slightly adapted when it was used with different columns. Similar selectivity is obtained not only on different C18 materials but also on C8 packings. The selectivity on poly(styrenedivinylbenzene) is less good. As method performance test a resolution test with benzylpenicillin was used. The robustness of this method was examined by applying a full factorial design to test the influence of the content of organic modifier in the mobile phase, the pH of the mobile phase, the concentration of buffer in the mobile phase and the temperature of the column. The results show that the method is robust. PMID- 16256707 TI - Arginine vasopressin transport and metabolism in the pigmented rabbit conjunctiva. AB - The purpose of this study was to evaluate the transepithelial transport and metabolism of arginine vasopressin (AVP) in the pigmented rabbit conjunctiva, both in the absence and presence of protease inhibitors. The apparent permeability coefficient, P(app), for 3H-AVP was determined in the modified Ussing chamber, and AVP metabolites were monitored by reversed phase HPLC using a C18 column. At 50 nM donor 3H-AVP, the P(app) in the mucosal-to-serosal (ms) direction was about five times higher than that in the opposite direction. Excess (0.1 mM) AVP decreased the P(app) for labelled AVP in the mucosal-to-serosal (ms) direction by about 50%. However, intact AVP transport showed neither concentration nor direction dependence. HPLC analysis revealed two subspecies of 3H-AVP in the receiver fluid and virtually no degradation products in the donor fluid following 3 h flux experiments. 3H-AVP transported in the ms direction underwent extensive hydrolysis (73%), which was decreased by 33% with mucosal application of 2 mM camostat mesylate (an aminopeptidase inhibitor) or by 27% with 0.5 mM leupeptin (a serine protease inhibitor). By contrast, 3H-AVP transported in the serosal-to-mucosal (sm) direction resulted in only 37% hydrolysis, and mucosal application of either inhibitor did not significantly affect the P(app) for intact AVP. These data suggest that intact AVP transport in the conjunctiva may be mediated mostly by passive diffusion and enzymatic degradation of AVP may be mediated by proteolytic enzymes present on the mucosal side of the conjunctiva. PMID- 16256708 TI - Enhancing effect of hydroxypropyl-beta-cyclodextrin on cutaneous penetration and activation of ethyl 4-biphenylyl acetate in hairless mouse skin. AB - The effect of hydroxypropyl-beta-cyclodextrin (HP-beta-CyD) on the cutaneous penetration and activation of ethyl 4-biphenylyl acetate (EBA), a prodrug of non steroidal anti-inflammatory drug 4-biphenylylacetic acid (BPAA), from hydrophilic ointment was investigated, using hairless mouse skin in vitro. When the hydrophilic ointment containing a complex of EBA with HP-beta-CyD was applied to the full-thickness skin, HP-beta-CyD facilitated the penetration of EBA into the skin, the conversion of EBA to BPAA in the epidermis and the transfer of BPAA to the receptor phase. Under the present condition, pre- and post-application of the ointment containing HP-beta-CyD onto the skin did not affect the cutaneous penetration of EBA and its activation. When the ointment containing the EBA:HP beta-CyD complex was applied to the skin, the flux of BPAA through the tape stripped skin was greater than that through the full-thickness skin, while the activation of the prodrug in the skin was slowed down by the tape-stripping. When propylene glycol was used as a vehicle, HP-beta-CyD no longer enhanced the cutaneous permeation of BPAA through the full-thickness skin. These results suggest that the enhancing effect of HP-beta-CyD on the cutaneous penetration of EBA would be ascribable largely to an increase in effective concentration of EBA in the ointment. Furthermore, the slow diffusion of EBA solubilized in HP-beta CyD through the stratum corneum, together with the vehicle effect, could make the prodrug more susceptible to the metabolic process that is active in the epidermis, eventually leading to the facilitated activation of the prodrug. PMID- 16256709 TI - Metabolite-derived artificial polymers designed for drug targeting, cell penetration and bioresorption. AB - The use of poly(beta-malic acid) and poly(l-lysine citramide) carriers to transport doxorubicin (Dox) within K562 myeloblastic cells was studied by taking advantage of laser microspectrofluorometry (L-MSF). This technique provided a means to monitor and to quantify the penetration of Dox molecules in the cytoplasm and in the nucleus of Dox-sensitive and Dox-resistant cells. Comparison was made between polymer-drug conjugates more or less hydrophobised by C2, C7 and C12 aliphatic substituents and by the Dox attached to the polymer backbone as pendent chains. Furthermore, a method was proposed to introduce a cleavable non peptidic spacer of the lactyllactyl-type between the poly(L-lysine citramide) backbone and the drug. It is shown that: Dox was released from the conjugates by a non-enzymatic route in the absence of cells, the Dox-uptake by cells was slower for the conjugates than for the free drug, and the hydrophobisation promoted the penetration of the released drug within the nucleus, even in the case of Dox resistant cells. However, no reversion of the resistance was observed. PMID- 16256711 TI - Regional organization of perinatal care. PMID- 16256710 TI - In vivo non-linear intestinal permeability of celiprolol and propranolol in conscious dogs: evidence for intestinal secretion. AB - The objective of this study was to investigate the absorption mechanism of celiprolol as a potential source of the drug's non-linear oral pharmacokinetics by determining its intestinal permeability as a function of concentration in vivo in dogs. Solutions of different celiprolol concentrations containing propranolol as an internal absorption marker were perfused through an isolated jejunal segment and samples were analyzed by an enantioselective HPLC method (Hartmann et al., J. Chromatogr., 496 (1989) 387-396). Permeability (P(eff) x 10(4) cm/s) of celiprolol increased significantly from 1.9-2.1 for the lower concentrations to 3.2 for the highest concentration, while the variability decreased. No statistical differences in the uptake between the two enantiomers were observed. Permeability of propranolol also increased significantly with increasing celiprolol concentrations, suggesting that propranolol might be utilizing the same carrier protein. In conclusion, the non-linear and variable oral pharmacokinetics of celiprolol might be due to a non-linear saturable, possibly secretion component in its uptake mechanism. PMID- 16256712 TI - Development of regionalized perinatal care. AB - Regionalized perinatal care was first advocated in Canada 35 years ago. Its development in the United States of America (from 1971), United Kingdom (from 1972) and Australia (from 1978), has been described. In all instances, the efforts and perseverance of visionary individuals were crucial in introducing the concepts to the medical profession, and in bringing the principles to national consciousness at a governmental level. Official endorsement of regionalized perinatal care by both the national professional bodies and the central and regional governments was necessary. An important milestone along the path of regionalized perinatal care was the establishment of national training programmes, and the recognition of maternal-fetal medicine and neonatal-perinatal medicine as respective subspecialties for obstetricians and paediatricians. The developmental process requires close collaboration among all three groups (individuals, professional bodies and government) to improve both the quality and availability of perinatal services to a geographically defined region. PMID- 16256713 TI - Regionalization of perinatal care in Europe. AB - This review describes European health policies related to the place of birth of very preterm babies, and the organizational context in which these policies were enacted using data from two European studies. It also compiles available information on the place of birth of very preterm babies from the published literature. In Europe, there is significant diversity in approaches to the provision of intensive care services for the small proportion of pregnant women and babies that need it, both in terms of health policies and the supply and characteristics of maternity and neonatal units. These diverse models in countries with similar levels of development and medical technology could offer an opportunity to understand how different organizational characteristics affect access to care, health outcomes and resource use. PMID- 16256714 TI - Regionalized perinatal care in North America. AB - The aim of this article is to familiarize the reader with the status of regionalized perinatal health care in North America, and specifically in the United States of America, using the Iowa regionalization model. The evolution of the regionalization movement in the late 1960s and early 1970s is reviewed. It is noted that the movement was largely without federal government intervention. The role of the March of Dimes in developing the first set of so-called national guidelines is chronicled. The Iowa model, utilizing some non-tertiary referral centres, is discussed in depth. This model included extensive outreach education for the entire state, and worked well largely because of the lack of competition to the major university teaching hospital located at the University of Iowa in Iowa City. This was not true for many other states and regions because competition did exist between tertiary centres. The impact of managed care systems on the overall role of regionalization is discussed. Suggestions are offered for controlling the impact of third-party payers (managed care) on the quality of perinatal health care. An idealized system of monitoring the impact of regionalization and monitoring the effect of managed care is detailed. Finally, the future of regionalization is discussed in the face of deregionalization in populous areas. The need for the best possible care as close to patients' homes as possible (regionalization) still seems apparent. PMID- 16256715 TI - Regionalized perinatal care in developing countries. AB - Neonatal-perinatal ill health and mortality are overwhelmingly a burden of the developing world. As many as 90% of births, 98% of fetal deaths and 98% of neonatal deaths occur in less developed countries. Regionalized perinatal services were introduced in developed countries when most neonatal mortality was confined to very-low-birthweight babies who required intensive perinatal care to survive. A large proportion of newborn morbidity and mortality in developing countries, however, continues to occur among full-term and moderate-sized low birthweight neonates who can be managed well in the community and at small hospitals. The model of regionalized perinatal care as practiced in developed countries is, at present, neither affordable nor relevant to the needs of many developing countries. It is possible to achieve considerably lower neonatal mortality rates in resource-poor settings by implementing home-based newborn care delivered by community health workers, and by promoting institutional perinatal care at simple facilities provided by trained midwives. PMID- 16256716 TI - Regionalized neonatal emergency transport. AB - This article reviews the components that facilitate an effective neonatal emergency transport network, and discusses the human resources required for safe transport, including a section focused on the option of an expanded role for the paramedic. In addition, the topics of transport equipment, communications, quality assurance, data management, family support and education are addressed in the context of a neonatal transport programme. Finally, elements involved in the organization of neonatal transport and transport issues pertaining to networking of neonatal medical care are highlighted and illustrated with reference to local experience in British Columbia. PMID- 16256717 TI - Regionalized long-term follow-up. AB - The importance of population-based long-term follow-up studies of geographically determined cohorts to evaluate the effectiveness, efficiency and availability of a regionalized perinatal-neonatal care programme is demonstrated by the Victorian Infant Collaborative Study Group. The survival and quality of survival of consecutively born extremely-low-birthweight infants below 1000 g or extremely preterm infants below 28 weeks' gestation in the state of Victoria were assessed up to 14 years of age over four distinctive eras: 1979-1989, 1985-1987, 1991-1992 and 1997. Both survival and quality-adjusted survival rates rose progressively in all birth weight and gestation subgroups, associated with progressively more such infants being born in level III perinatal centres. Cost-effectiveness and cost utility ratios remained stable overall, with efficiency gains in the smaller infants over time. Regionalized long-term follow-up provides unique information that is not available from institution-based studies, which is vital to the regional organization of perinatal-neonatal care. PMID- 16256718 TI - Regional quality assessment in perinatal care. AB - Quality assessment is essential in every sector of health care and, in modern regionalized perinatal care, continuous data should be collected at all levels to give a stable basis for this activity. The discussion of definitions and choice of indicators is in itself an activity that will increase awareness of quality. Modern computer facilities will simplify data storage and analysis, but do not change the need to use a limited number of well-validated and appropriate variables. PMID- 16256720 TI - Interpretation of the paediatric chest X-ray. AB - Interpretation of the paediatric chest X-ray may appear intimidating at first but knowledge of a few basic rules and an understanding of how the radiographic appearance may be influenced by age and technique will help the clinician arrive at the correct diagnosis in many cases. A structured and logical approach to X ray interpretation is described, drawing attention to the abnormal signs that should be sought. A limited differential diagnosis is given for some of the more frequent radiological abnormalities seen in children. PMID- 16256719 TI - Regionalized perinatal education. AB - Despite changes in the organization and financing of healthcare delivery, and dramatic increases in the number and distribution of perinatal facilities and professionals over the past three decades, there remains a continuing need for effective and efficient regionalized perinatal outreach education programmes. Both the organizers and the participants should be multidisciplinary and include both inpatient and outpatient providers. Content should be restricted to issues relevant to participants' practice, and include topics ranging from preconception to postpartum and early infant care. There are various effective formats, but consideration should be given to reaching as many providers as possible simultaneously within a given facility, minimizing expense and economizing on participants' time. Evaluation strategies range from assessment of immediate outcomes, which generally examine programme process, to ultimate outcomes, which measure changes in patient care and patient health. PMID- 16256723 TI - Case 2: Assessment: cardiac enlargement. PMID- 16256724 TI - Case 4: Assessment: chest X-ray and V/Q scan. PMID- 16256725 TI - Case 3: Assessment: A small girl with persistent purulent rhinorrhoea and coughing. PMID- 16256726 TI - Case 5: Assessment: chest X-ray and V/Q scan. PMID- 16256728 TI - Case 2: assessment. Postural-dependent stridor. PMID- 16256730 TI - Pak GITs to Aurora-A. AB - The protein kinase Aurora-A orchestrates key aspects of cell division, but its regulation has remained a major puzzle. That puzzle is now one step closer to being solved thanks to a recent paper by Zhao et al. appearing in the October 28(th) issue of Molecular Cell. In this work, the authors show that p21-activated kinases (Paks), previously best known for their role in cytoskeletal and transcriptional signaling pathways, also play an important role in centrosome dynamics via phosphorylation and activation of Aurora-A. PMID- 16256731 TI - A new notch in the HIF belt: how hypoxia impacts differentiation. AB - In the current issue of Developmental Cell, work by Gustafsson and coworkers demonstrates that hypoxia synergizes with Notch to inhibit differentiation of myogenic and neural precursor cells. This effect requires a newly described interaction between the transcriptionally active form of HIF-1alpha and the intracellular domain of Notch. PMID- 16256732 TI - Tracing the sources of cellular variation. AB - As the adage says, variety is the spice of life, and despite our best attempts, cells, even those with the same genome, never seem to behave the same. By combining mathematical and experimental analyses, Colman-Lerner and colleagues propose, in a recent issue of Nature, a method to delicately unravel the sources of this variation. Applying their technique to the pheromone response in budding yeast, they show that much of the observed variation originates from cell cycle effects and is dependent on levels of pathway input. PMID- 16256733 TI - A ring-like template for abscission. AB - In the October 7th issue of Cell, it is shown that a ring-like structure containing the centrosomal protein centriolin acts as a local recruitment site for the membrane fusion machinery that controls abscission. PMID- 16256734 TI - Dynamics of endocytic vesicle creation. AB - Clathrin-mediated endocytosis is the main path for receptor internalization in metazoans and is essential for controlling cell integrity and signaling. It is driven by a large array of protein and lipid interactions that have been deciphered mainly by biochemical and genetic means. To place these interactions into context, and ultimately build a fully operative model of endocytosis at the molecular level, it is necessary to know the kinetic details of the role of each protein in this process. In this review, we describe the recent efforts made, by using live cell imaging, to define clear steps in the formation of endocytic vesicles and to observe the recruitment of key proteins during membrane invagination, the scission of a newly formed vesicle, and its movement away from the plasma membrane. PMID- 16256735 TI - SoxE factors function equivalently during neural crest and inner ear development and their activity is regulated by SUMOylation. AB - Sox9 and the closely related factor Sox10 are essential for the formation of neural crest precursor cells, and play divergent roles in the process by which these cells are subsequently directed to form specific derivatives. These group E Sox factors have also been implicated in the development of the vertebrate inner ear. Despite their importance, however, the mechanisms that allow SoxE proteins to regulate such a diverse range of cell types have remained poorly understood. Here we demonstrate that during vertebrate development, the activities of individual SoxE factors are well conserved and are regulated by SUMOylation. We show that SoxE mutants that cannot be SUMOylated, or that mimic constitutive SUMOylation, are each able to mediate a subset of the diverse activities characteristic of wild-type SoxE proteins. These findings provide important mechanistic insight into how the activity of widely deployed developmental regulatory proteins can be directed to specific developmental events. PMID- 16256736 TI - A role for SIR-2.1 regulation of ER stress response genes in determining C. elegans life span. AB - C. elegans SIR-2.1, a member of the Sir-2 family of NAD(+)-dependent protein deacetylases, has been shown to regulate nematode aging via the insulin/IGF pathway transcription factor daf-16. Treatment of C. elegans with the small molecule resveratrol, however, extends life span in a manner fully dependent upon sir-2.1, but independent of daf-16. Microarray analysis of worms treated with resveratrol demonstrates the transcriptional induction of a family of genes encoding prion-like glutamine/asparagine-rich proteins involved in endoplasmic reticulum (ER) stress response to unfolded proteins. RNA interference of abu-11, a member of this ER stress gene family, abolishes resveratrol-mediated life span extension, and overexpression of abu-11 extends the life span of transgenic animals. Furthermore, SIR-2.1 normally represses transcription of abu-11 and other ER stress gene family members, indicating that resveratrol extends life span by inhibiting sir-2.1-mediated repression of ER stress genes. Our findings demonstrate that abu-11 and other members of its ER stress gene family are positive determinants of C. elegans life span. PMID- 16256738 TI - HMGCoA reductase potentiates hedgehog signaling in Drosophila melanogaster. AB - Drosophila HMGCoA reductase (hmgcr) catalyzes the biosynthesis of a mevalonate precursor for isoprenoids and has been implicated in the production of a signal by the somatic gonadal precursor cells (SGPs) that attracts migrating germ cells. Here, we show that hmgcr functions in the hedgehog (hh) signaling pathway. When hmgcr activity is reduced, high levels of Hh accumulate in hh-expressing cells in each parasegment, while the adjacent "Hh-receiving" cells cannot sustain wg expression and fail to relocalize the Smoothened (Smo) receptor. Conversely, ectopic Hmgcr upregulates Hh signaling when it is produced in hh-expressing cells, but has no effect when produced in the receiving cells. These findings suggest that Hmgcr might orchestrate germ cell migration by promoting the release and/or transport of Hh from the SGPs. Consistent with this model, there are substantial germ cell migration defects in trans combinations between hmgcr and mutations in different components of the hh pathway. PMID- 16256737 TI - Hypoxia requires notch signaling to maintain the undifferentiated cell state. AB - In addition to controlling a switch to glycolytic metabolism and induction of erythropoiesis and angiogenesis, hypoxia promotes the undifferentiated cell state in various stem and precursor cell populations. Here, we show that the latter process requires Notch signaling. Hypoxia blocks neuronal and myogenic differentiation in a Notch-dependent manner. Hypoxia activates Notch-responsive promoters and increases expression of Notch direct downstream genes. The Notch intracellular domain interacts with HIF-1alpha, a global regulator of oxygen homeostasis, and HIF-1alpha is recruited to Notch-responsive promoters upon Notch activation under hypoxic conditions. Taken together, these data provide molecular insights into how reduced oxygen levels control the cellular differentiation status and demonstrate a role for Notch in this process. PMID- 16256739 TI - Canonical Wnt signaling and its antagonist regulate anterior-posterior axis polarization by guiding cell migration in mouse visceral endoderm. AB - The mouse embryonic axis is initially formed with a proximal-distal orientation followed by subsequent conversion to a prospective anterior-posterior (A-P) polarity with directional migration of visceral endoderm cells. Importantly, Otx2, a homeobox gene, is essential to this developmental process. However, the genetic regulatory mechanism governing axis conversion is poorly understood. Here, defective axis conversion due to Otx2 deficiency can be rescued by expression of Dkk1, a Wnt antagonist, or following removal of one copy of the beta-catenin gene. Misexpression of a canonical Wnt ligand can also inhibit correct A-P axis rotation. Moreover, asymmetrical distribution of beta-catenin localization is impaired in the Otx2-deficient and Wnt-misexpressing visceral endoderm. Concurrently, canonical Wnt and Dkk1 function as repulsive and attractive guidance cues, respectively, in the migration of visceral endoderm cells. We propose that Wnt/beta-catenin signaling mediates A-P axis polarization by guiding cell migration toward the prospective anterior in the pregastrula mouse embryo. PMID- 16256740 TI - BMP signaling is required for controlling somatic stem cell self-renewal in the Drosophila ovary. AB - BMP signaling is essential for promoting self-renewal of mouse embryonic stem cells and Drosophila germline stem cells and for repressing stem cell proliferation in the mouse intestine and skin. However, it remains unknown whether BMP signaling can promote self-renewal of adult somatic stem cells. In this study, we show that BMP signaling is necessary and sufficient for promoting self-renewal and proliferation of somatic stem cells (SSCs) in the Drosophila ovary. BMP signaling is required in SSCs to directly control their maintenance and division, but is dispensable for proliferation of their differentiated progeny. Furthermore, BMP signaling is required to control SSC self-renewal, but not survival. Moreover, constitutive BMP signaling prolongs the SSC lifespan. Therefore, our study clearly demonstrates that BMP signaling directly promotes SSC self-renewal and proliferation in the Drosophila ovary. Our work further suggests that BMP signaling could promote self-renewal of adult stem cells in other systems. PMID- 16256741 TI - Phosphorylation of ACAP1 by Akt regulates the stimulation-dependent recycling of integrin beta1 to control cell migration. AB - Components of intracellular signaling that mediate the stimulation-dependent recycling of integrins are being identified, but key transport effectors that are the ultimate downstream targets remain unknown. ACAP1 has been shown recently to function as a transport effector in the cargo sorting of transferrin receptor (TfR) that undergoes constitutive recycling. We now show that ACAP1 also participates in the regulated recycling of integrin beta1 to control cell migration. However, in contrast to TfR recycling, the role of ACAP1 in beta1 recycling requires its phosphorylation by Akt, which is, in turn, regulated by a canonical signaling pathway. Disrupting the activities of either ACAP1 or Akt, or their assembly with endosomal beta1, inhibits beta1 recycling and cell migration. These findings advance an understanding of how integrin recycling is achieved during cell migration, and also address a basic issue of how intracellular signaling can interface with transport to achieve regulated recycling. PMID- 16256742 TI - Efficient protein trafficking requires trailer hitch, a component of a ribonucleoprotein complex localized to the ER in Drosophila. AB - Translational control of localized messenger mRNAs (mRNAs) is critical for cell polarity, synaptic plasticity, and embryonic patterning. While progress has been made in identifying localization factors and translational regulators, it is unclear how broad a role they play in regulating basic cellular processes. We have identified Drosophila trailer hitch (tral) as a gene that is required for the proper secretion of the dorsal-ventral patterning factor Gurken, as well as the vitellogenin receptor Yolkless. Surprisingly, biochemical purification of Tral revealed that it is part of a large RNA-protein complex that includes the translation/localization factors Me31B and Cup as well as the mRNAs for endoplasmic reticulum (ER) exit site components. This complex is localized to subdomains of the ER that border ER exit sites. Furthermore, tral is required for normal ER exit site formation. These findings raise exciting new possibilities for how the mRNA localization machinery could interface with the classical secretory pathway to promote efficient protein trafficking in the cell. PMID- 16256743 TI - The Drosophila tumor suppressor vps25 prevents nonautonomous overproliferation by regulating notch trafficking. AB - Cell-cell signaling coordinates proliferation of metazoan tissues during development, and its alteration can induce malignant transformation. Endocytosis regulates signaling by controlling the levels and activity of transmembrane receptors, both prior to and following ligand engagement. Here, we identify Vps25, a component of the ESCRT machinery that regulates endocytic sorting of signaling receptors, as an unconventional type of Drosophila tumor suppressor. vps25 mutant cells undergo autonomous neoplastic-like transformation, but they also stimulate nonautonomous cell proliferation. Endocytic trafficking defects in vps25 cells cause endosomal accumulation of the signaling receptor Notch and enhanced Notch signaling. Increased Notch activity leads to ectopic production of the mitogenic JAK-STAT pathway ligand Unpaired, which is secreted from mutant cells to induce overproliferation of the surrounding epithelium. Our data show that defects in endocytic sorting can both transform cells and, through heterotypic signaling, alter the behavior of neighboring wild-type tissue. PMID- 16256744 TI - Mutations in erupted, the Drosophila ortholog of mammalian tumor susceptibility gene 101, elicit non-cell-autonomous overgrowth. AB - The reproducible pattern of organismal growth during metazoan development is the product of genetically controlled signaling pathways. Patterned activation of these pathways shapes developing organs and dictates overall organismal shape and size. Here, we show that patches of tissue that are mutant for the Drosophila Tsg101 ortholog, erupted, cause dramatic overproliferation of adjacent wild-type tissue. Tsg101 proteins function in endosomal sorting and are required to incorporate late endosomes into multivesicular bodies. Drosophila cells with impaired Tsg101 function show accumulation of the Notch receptor in intracellular compartments marked by the endosomal protein Hrs. This causes increased Notch mediated signaling and ectopic expression of the Notch target gene unpaired (upd), which encodes the secreted ligand of the JAK-STAT pathway. Activation of JAK-STAT signaling in surrounding wild-type cells correlates with their overgrowth. These findings define a pathway by which changes in endocytic trafficking can regulate tissue growth in a non-cell-autonomous manner. PMID- 16256747 TI - Dietetics and disability. PMID- 16256745 TI - Tumor suppressor properties of the ESCRT-II complex component Vps25 in Drosophila. AB - We have found that the Drosophila gene vps25 possesses several properties of a tumor suppressor. First, vps25 mutant cells activate Notch and Dpp receptor signaling, inducing ectopic organizers in developing eyes and limbs and consequent overproliferation of both mutant and nearby wild-type cells. Second, as the mutant cells proliferate, they lose their epithelial organization and undergo apoptosis. Strikingly, when apoptosis of mutant cells is blocked, tumor like overgrowths are formed that are capable of metastasis. vps25 encodes a component of the ESCRT-II complex, which sorts membrane proteins into multivesicular bodies during endocytic trafficking to the lysosome. Activation of Notch and Dpp receptor signaling in mutant cells results from an endocytic blockage that causes accumulation of these receptors and other signaling components in endosomes. These results highlight the importance of endocytic trafficking in regulating signaling and epithelial organization and suggest a possible role for ESCRT components in human cancer. PMID- 16256748 TI - The obesity epidemic: being part of the solution. PMID- 16256749 TI - Roles of vitamin D, n-3 polyunsaturated fatty acid, and soy isoflavones in bone health. PMID- 16256750 TI - Dietary challenges of new Americans. PMID- 16256751 TI - Professional wills: what's that all about? PMID- 16256752 TI - Knowing when to hold and when to fold in advocacy. PMID- 16256753 TI - Clarification of competitive foods. PMID- 16256755 TI - The role of diet and lifestyle modifications in the statin era. PMID- 16256756 TI - Unique dietary patterns and chronic disease risk profiles of adult men: the Framingham nutrition studies. AB - OBJECTIVE: To identify the dietary patterns of adult men and examine their relationships with nutrient intake and chronic disease risk over long-term follow up. DESIGN/SUBJECTS: Baseline 145-item food frequency questionnaires from 1,666 Framingham Offspring-Spouse cohort men were used to identify comprehensive dietary patterns. Independent 3-day dietary records at baseline and 8 years later provided estimates of subjects' nutrient intake by dietary pattern. Chronic disease risk factor status was compared at baseline and 16-year follow-up across all male dietary patterns. STATISTICAL ANALYSES: Cluster analysis was applied to food frequency data to identify non-overlapping male dietary patterns. Analysis of covariance and logistic regression were used to compare nutrient intake, summary nutritional risk scores, and chronic disease risk status at baseline and follow-up by male dietary pattern. RESULTS: Five distinct and comprehensive dietary patterns of Framingham Offspring-Spouse men were identified and ordered according to overall nutritional risk: Transition to Heart Healthy, Higher Starch, Average Male, Lower Variety, and Empty Calories. Nutritional risk was high and varied by dietary pattern; key nutrient contrasts were stable over 8 year follow-up. Chronic disease risk also varied by dietary pattern and specific subgroup differences persisted over 16 years, notably rates of overweight/obesity and smoking. CONCLUSIONS: Quantitative cluster analysis applied to food frequency questionnaire data identified five distinct, comprehensive, and stable dietary patterns of adult Framingham Offspring-Spouse cohort men. The close associations between the dietary patterns, nutritional risk, and chronic disease profiles of men emphasize the importance of targeted preventive nutrition interventions to promote health in the male population. PMID- 16256757 TI - Relationship of fruit and vegetable consumption in middle-aged men to medicare expenditures in older age: the Chicago Western Electric Study. AB - BACKGROUND: High fruit and vegetable intake is associated with lower risk of hypertension, cardiovascular disease, and cancer. Little is known about the relationship of fruit and vegetable intake to health care expenditures. OBJECTIVE: Examine whether fruit and vegetable intake among middle-aged adults is related to Medicare charges-total, cardiovascular disease, cancer-related-in older age. DESIGN: Participants were grouped into one of three strata according to fruit and vegetable intake, determined from detailed dietary history (1958 1959): less than 14 cups per month, 14 to 42 cups per month, or more than 42 cups per month. Combined intake was classified as low, medium, or high. Medicare claims data (1984-2000) were used to estimate mean annual spending for eligible surviving participants (65 years and older) from the Chicago Western Electric Study: 1,063 men age 40 to 55 and without coronary heart disease, diabetes, and cancer at baseline (1957-1958). Cumulative charges before death (n = 401) were also calculated. RESULTS: Higher fruit and fruit plus vegetable intakes were associated with lower mean annual and cumulative Medicare charges (P values for trend .019 to .862). For example, with adjustment for baseline age, education, total energy intake, and multiple baseline risk factors, annual cardiovascular disease-related charges were 3,128 dollars vs 4,223 dollars for men with high vs low intake of fruit plus vegetables. Corresponding figures were 1,352 dollars vs 1,640 dollars for cancer-related charges and 10,024 dollars vs 12,211 dollars for total charges. Results were generally similar for vegetable intake. CONCLUSION: These findings, albeit mostly not statistically significant, suggest that for men high intake of fruits and fruits plus vegetables earlier in life has potential not only for better health status but also for lower health care costs in older age. PMID- 16256758 TI - Differences in fruit and vegetable intake among categories of dietary supplement users. AB - BACKGROUND: People who choose to take dietary supplements are often classified as having a healthful lifestyle; however, it is probable that several health behavior patterns exist among users. OBJECTIVE: Data from the North Carolina Strategies to Improve Diet, Exercise, and Screening study (N = 727) were used to identify and describe five different categories of dietary supplement use and evaluate how these categories are associated with both quantity and quality of fruit and vegetable consumption. DESIGN: Five nonoverlapping dietary supplement use categories were created and descriptive demographic statistics were compared. Least-squares means were calculated for knowledge and fruit and vegetable quality and quantity measures. Logistic regression was performed to calculate adjusted odds ratios to examine associations among the dietary supplement use categories and the fruit and vegetable measures, using nonusers as the reference category. RESULTS: Demographic profiles and dietary intake varied among the dietary supplement categories. People reporting both a multivitamin/multimineral and a single supplement (Multi Plus category) and any nonvitamin/nonmineral products (Herbals category) were more likely to be consuming more vegetables and higher quality fruits and vegetables than those not taking any dietary supplements (Nonusers category), whereas people taking multivitamins/multiminerals only (Multis category) had patterns that were more consistently similar to those not taking any dietary supplements (Nonusers). CONCLUSIONS: Study participants exhibited dietary supplement use patterns that were associated with differences in fruit and vegetable consumption. Simply characterizing people as users and nonusers will not capture critical demographic and dietary differences and will likely further cloud investigations of diet-disease relationships. PMID- 16256759 TI - Including dietary supplements in nutrition assessment. PMID- 16256760 TI - Comparability and reliability of paper- and computer-based measures of psychosocial constructs for adolescent fruit and vegetable and dietary fat intake. AB - BACKGROUND: This study investigated the comparability and reliability of computer and paper-based measures of psychosocial constructs related to fruit and vegetable and dietary fat intake among adolescents. METHODS: An ethnically diverse sample of 76 adolescents was studied (mean age 13 years). Scales measured use of change strategies, self-efficacy, decisional balance, family influences, and peer influences separately for the two dietary outcomes. RESULTS: Comparability analyses indicated that responses for each of the 12 diet-related scales were not significantly different between the computer- and paper-based surveys. Internal consistencies were generally high (alpha from .61 to .97) with slightly better reliability on the computer- vs paper-based surveys. Test-retest reliabilities were adequate to good for most multiple-item scales (interclass correlation coefficients from .43 to .85 and .48 to .90 for paper and computer formats, respectively). CONCLUSIONS: Computer- and paper-based measures of psychosocial constructs are appropriate and ready for use in either format for studies of dietary behaviors in young people. PMID- 16256761 TI - Dietary patterns of young, low-income US children. AB - OBJECTIVE: The purpose of this study was to identify dietary patterns of young, low-income US children, describe differences in diet quality between identified patterns, and make targeted food recommendations to improve diet quality. DESIGN: Dietary patterns were assessed using dietary variables from the Pyramid Servings Database within the Continuing Survey of Food Intakes by Individuals 1994-1996, 1998. Healthy Eating Index (HEI) scores were used to validate identified dietary patterns. SUBJECTS/SETTING: Two subsamples of low-income children, aged 2 to 3 years (n = 1,242) and 4 to 8 years (n = 1,506), were selected from the Continuing Survey of Food Intakes by Individuals data. STATISTICAL ANALYSIS: Cluster analysis was performed to determine unique dietary patterns within the two subsamples. Linear regression analyses were used to compare energy intake, discretionary fat, added sugars, and HEI scores across cluster groups. Descriptive statistics were computed for each cluster. RESULTS: Cluster analysis identified six and seven distinct dietary patterns for the younger and older children, respectively. Four patterns were similar for both age groups. For the 2 to 3-year-old children, energy intake, overall HEI scores, and nine of the 10 HEI component scores differed among the four most prevalent dietary patterns. Among the older children, energy intake and six of the HEI component scores differed between the four most prevalent clusters but not overall HEI. CONCLUSIONS: Neither age group had a cluster of children who followed a balanced/moderate diet pattern consistent with Food Guide Pyramid recommendations. Children consuming almost every pattern identified could benefit by reducing added sugars and discretionary fat and increasing low-fat, low-sugar options from the vegetables, fruits, meat, and milk groups. PMID- 16256762 TI - Acceptability of stage-tailored newsletters about fruits and vegetables by young adults. AB - The purpose of this study was to evaluate the acceptability of stage-tailored newsletters as a communication means for promoting fruit and vegetable intake by young adults. Qualitative interviews were conducted to gather young adults' likes and dislikes about stage-tailored newsletters on fruits and vegetables. A total of 246 interviews were completed by young adults between the ages of 18 and 24 years for focus group and/or in-depth interviews. Prototype newsletters containing stage-matched messages were designed to increase motivational readiness to increase fruit and vegetable intake based on the Transtheoretical Model. Transcripts were grouped by stage of change and examined to identify major themes or topics. Results showed that young adults in each of the three stages responded positively to the tailored newsletters. Subjects provided feedback on newsletter likes/dislikes, benefits/barriers to eating fruits and vegetables, and strategies for promoting change. This study provides an example of the incorporation of key concepts from the Transtheoretical Model in development of stage-tailored newsletters for young adults. PMID- 16256763 TI - Food type, food preparation, and competitive food purchases impact school lunch plate waste by sixth-grade students. AB - Because plate waste reduces nutritional benefits of school lunches, the objective of this study was to determine the factors that influence waste. Lunches of 743 sixth graders in three middle schools were photographed after students made food selections and after eating. Sample items were purchased to determine initial portion weights. The Statistical Package for Social Sciences software (Base 10.0, 1995, SPSS Inc, Chicago, IL) was used for statistical analysis. Preparation method influenced acceptance rates and plate waste: Whole apples had lower acceptance (23%) and greater waste (62%) compared with applesauce (37% acceptance, 23% waste). Mashed potatoes and heated fries had similar acceptance (approximately 69%), but mashed potatoes were wasted less (19% vs 33%, respectively). Students who purchased competitive food items with lunch (n = 250) had significantly greater waste of fruits (52% vs 36%, respectively, P = .0001), grain products (26% vs 14%, respectively, P = .009), meats (25% vs 16%, respectively, P = .015), and mixed dishes (30% vs 18%, respectively, P = .0001). Changing preparation methods and limiting availability of competitive food items may reduce plate waste. PMID- 16256765 TI - Salad bars and fruit and vegetable consumption in elementary schools: a plate waste study. AB - The object of this study was to determine whether students attending schools with self-service salad bars consume a greater amount of fruits and vegetables compared with students using preportioned servings and to evaluate the relationship between number of items offered and fruit and vegetable consumption. Two hundred ninety-four students in first through fifth grade were randomly selected from two schools with salad bars and two with preportioned servings. Weights of fruit and vegetable items were measured pre- and postconsumption and interobserver agreement +/-1 g was > or =95%. Presence of a salad bar was not associated with greater fruit and vegetable consumption. Fruit and vegetable consumption was positively related to the number of fruit and vegetable items offered at salad bars (P < .05), adjusting for sex and grade. Fruit and vegetable variety was associated with elementary school-age children's fruit and vegetable consumption when using salad bars. PMID- 16256766 TI - A "contract for change" increases produce consumption in low-income women: a pilot study. AB - This study determined whether a "Contract for Change" goal-setting exercise enhanced the effectiveness of the Expanded Food and Nutrition Education/Food Stamp Nutrition Education programs to increase produce consumption in low-income (<130% of poverty) women after 4 weeks. Thirty-eight participants were randomized in this three-group parallel arm study: (a) control group participants received life-skills lessons, (b) the education group received the Expanded Food and Nutrition Education/Food Stamp Nutrition Education "Food Guide Pyramid" lessons, and (c) the contract group also received the "Food Guide Pyramid" series and completed a "Contract for Change." It was hypothesized that the contract group would have the greatest increases in advancement toward dietary change and produce consumption. Compared with controls, the contract group significantly moved toward acceptance of vegetable consumption (P < or = .05). Compared with the education group, the contract group significantly increased fruit consumption. Results suggest that nutrition professionals can effectively use goal-setting to assist low-income populations with dietary change. PMID- 16256764 TI - Adolescents' low-carbohydrate-density diets are related to poorer dietary intakes. AB - This study was undertaken to assess how low-carbohydrate-density diets below the acceptable macronutrient distribution range relate to food and micronutrient intake and sociodemographic and health-related characteristics. The multistage stratified cluster design in the 1990 Ontario Health Survey was used. There were 5,194 subjects, 12 to 18 years of age, in sampled households. Dietary data were collected via a food frequency questionnaire. Low-carbohydrate-density diets were consumed by 27.6% of males and 24.1% of females. Low-carbohydrate-density diets were related (P < .05) to reduced sufficiency of vegetables and fruit and higher consumption of meat and alternatives and added fats. The low-carbohydrate-density diet resulted in intakes lower in vitamin C and fiber and higher in cholesterol and total fat. The low-carbohydrate-density diet was directly associated with being Canadian-born (odds ratio [OR] = 1.78, 95% confidence interval [CI] = 1.27 to 2.50), overweight status (OR = 1.27, 95% CI = 1.02 to 1.57), smoking (OR = 1.53, 95% CI = 1.23 to 1.90), alcohol use (OR = 1.46, 95% CI = 1.21 to 1.75), and poorer self-rated health (OR = 1.47, 95% CI = 1.01 to 2.14). Use of the acceptable macronutrient distribution range identified adolescents with low carbohydrate-density diets whose food choices and nutrient intake may impact negatively on short- and long-term health. PMID- 16256768 TI - Dietetic management of pediatric overweight: development and description of a practical and evidence-based behavioral approach. PMID- 16256767 TI - California teachers perceive school gardens as an effective nutritional tool to promote healthful eating habits. AB - This study assessed elementary school teachers' perceived attitudes and barriers associated with school gardens, as well as the purpose and use of gardens in schools, specifically in relation to the link between gardens and nutrition. The questionnaire was mailed to California fourth-grade teachers at schools with gardens (N = 1,665). The response rate was 36% (n = 592). Teachers perceived the garden to be somewhat to very effective at enhancing academic performance, physical activity, language arts, and healthful eating habits. Nutrition was taught with the use of the garden by 47% of responding teachers. This research provides evidence for needed standards-based curricula materials and teacher training in relation to gardening and nutrition. The results from this study will contribute to development of needed resources and methods by which to encourage the use of gardens and nutrition education in schools. PMID- 16256769 TI - Does the n-3 fatty acid alpha-linolenic acid have an impact on coronary heart disease? PMID- 16256770 TI - Tricuspid valve replacement with a cryopreserved pulmonary homograft. PMID- 16256771 TI - Nutritional status in patients on left ventricular assist device support. PMID- 16256773 TI - Religion, politics...deep hypothermic circulatory arrest. PMID- 16256774 TI - A comparison of outcomes of mitral valve repair for degenerative disease with posterior, anterior, and bileaflet prolapse. AB - OBJECTIVE: We sought to compare the clinical and echocardiographic outcomes of mitral valve repair for mitral regurgitation in patients with degenerative disease of the mitral valve with posterior, anterior, or bileaflet prolapse. METHODS: Patients underwent operations from 1981 through 2001: 359 had posterior (mean age, 60.4 years), 92 had anterior (mean age, 53.3 years), and 250 had bileaflet (means age, 56.4 years) prolapse. Patients with anterior prolapse were younger (P = .04) and had more associated aortic valve disease (P = .02), particularly bicuspid aortic valve disease (P < .001). Anterior prolapse was corrected by using chordal replacement with Gore-Tex sutures in most patients, but early on in this series, leaflet resection, chordal shortening, and chordal transfer were also used. Echocardiograms were done annually, and clinical follow up was complete at a mean of 6.9 +/- 4.0 years (range, 0-23 years). RESULTS: The overall survival at 12 years was 75% +/- 5%, with no difference among the posterior, anterior, and bileaflet prolapse groups (P = .3). The freedom from reoperation at 12 years was 96% +/- 2% for posterior, 88% +/- 4% for anterior, and 94% +/- 2% for bileaflet prolapse (P = .019). Anterior prolapse was the only independent predictor of reoperation. The freedom from moderate or severe mitral regurgitation at 12 years was 80% +/- 4% for posterior, 65% +/- 8% for anterior, and 67% +/- 6% for bileaflet prolapse (P = .001). Anterior and bileaflet prolapse, age, ejection fraction of less than 40%, and aortic valve disease were independent predictors of recurrent moderate or severe mitral regurgitation. CONCLUSIONS: The pathophysiology of mitral regurgitation affects the durability of mitral valve repair for degenerative disease, and the results of posterior prolapse are better than those of anterior and bileaflet prolapse. This study indicates that rates of reoperation underscore the rates of failure of mitral valve repair. PMID- 16256775 TI - Residual high incidence of ventricular arrhythmias after left ventricular reconstructive surgery. AB - OBJECTIVE: Left ventricular reconstruction is performed in patients with ischemic cardiomyopathy and akinetic or dyskinetic left ventricular regions. These patients may remain at risk for malignant ventricular arrhythmias and hence may benefit from prophylactic implantable cardioverter-defibrillators. Specific guidelines for electrophysiologic testing and implantable cardioverter defibrillator implantation in patients undergoing left ventricular reconstruction are lacking. We aimed to assess the residual risk and timing of ventricular arrhythmias after left ventricular reconstruction to determine whether electrophysiologic risk stratification or implantable cardioverter-defibrillator implantation can be safely deferred. METHODS: Data were prospectively gathered on 217 consecutive patients with left ventricular ejection fractions less than 40% undergoing left ventricular reconstruction at our institution from 1997 to 2002. Patients were divided into 3 groups: group 1, implantable cardioverter defibrillator present before surgery; group 2, implantable cardioverter defibrillator implanted early after surgery; and group 3, no implantable cardioverter-defibrillator implanted. End points were all-cause mortality (censored for cardiac transplantation) and appropriate implantable cardioverter defibrillator therapies. RESULTS: Of 217 patients (mean age, 61 +/- 10 years [mean +/- SD]), survival after a median follow-up of 381 days was 90%. Electrophysiologic studies successfully identified patients at low risk. Appropriate implantable cardioverter-defibrillator therapies occurred in 20% of group 1 and 12% of group 2. The median time to the first implantable cardioverter defibrillator therapy from the time of left ventricular reconstruction was 43 days, and most first therapies (67%) occurred within the first 63 days. CONCLUSIONS: The early event rates (occurring in the first 90 days after left ventricular reconstruction) support the use of predischarge electrophysiologic studies, implantation of implantable cardioverter-defibrillators before discharge from the hospital, or both. PMID- 16256776 TI - The identification and development of Canadian coronary artery bypass graft surgery quality indicators. AB - OBJECTIVE: The study objective was to develop quality indicators for coronary artery bypass graft surgery that relate to quality of care, associate with preventable death, and could be reported on performance reports. METHODS: A comprehensive list of quality indicators was collected from quality improvement organizations including the Society For Thoracic Surgery, Northern New England Cardiovascular Disease Study Group, and Veteran's Affairs System. Indicators were collated from practice guidelines from the American College of Cardiology and the American Heart Association. A MEDLINE search using the keywords "quality indicators" and "coronary bypass" was completed. A 17-member multidisciplinary international expert panel was assembled, who voted using a 2-step Delphi process regarding association with quality of care, risk adjustment, association with preventable death, and inclusion on performance reports. RESULTS: A total of 149 quality indicators were examined. This list was distilled to 33 indicators related to quality of care, 10 indicators that could be adequately risk adjusted, 34 indicators related to preventable death, and 18 indicators to be included on performance reports. These selected indicators consisted of 19 outcome variables, 23 process of care variables, and 4 structure variables. The quality indicators believed to be useful on a Canadian institutional coronary artery bypass graft surgery report card included the following: 30-day mortality, in-hospital mortality, electrocardiographic myocardial infarction, red cell transfusion, allogeneic blood product transfusion, deep sternal wound infection, postoperative stroke, postoperative dialysis, intensive care unit readmission, intensive care unit length of stay, ventilation time, repeat cardiac operation, repeat surgery with cardiopulmonary bypass, repeat revascularization, waiting time to surgery, completion of surgery within a recommended waiting time, use of left internal thoracic artery graft, and institutional volume. CONCLUSIONS: This set of consensus quality indicators can be used as a standard list to be monitored by providers of coronary artery bypass graft surgery in an effort to continuously evaluate and improve their performance. PMID- 16256777 TI - Single-suture line placement of a pericardial stentless valve. AB - OBJECTIVE: Implantation of bioprostheses in the supra-annular position with the single suture line was first applied by O'Brien to porcine stentless valves. The aim of this study was to evaluate the clinical performance of the Pericarbon Freedom stentless bioprosthesis (Sorin Biomedica Cardio, Saluggia, Italy) implanted in supra-annular position with the single-suture line technique. The single-suture approach for the Pericarbon Freedom stentless bioprosthesis is obtained by trimming away all the extra tissue of the valve inflow side and scalloping the outflow side. METHODS: Between February 2002 and August 2004, a total of 65 consecutive patients at our institution (48% male, mean age 69 +/- 12 years) underwent aortic valve replacement with Pericarbon Freedom stentless bioprostheses implanted with a single suture line. Most recurrent etiology was senile degeneration (80%). Pericarbon Freedom 25-mm and 27-mm valves were the most frequently implanted. Thirty patients had concomitant procedures (mainly coronary artery bypass grafting, 16 patients). Overall crossclamp time was 76 +/- 21 minutes. RESULTS: All patients survived intervention. One patient died early of multiorgan failure (postoperative day 16). There were 4 early non-valve related complications and no late complications at a mean follow-up of 491 +/- 270 days. Four patients showed trivial central prosthetic regurgitation at intraoperative transesophageal echocardiography; among these cases, only 1 was confirmed at 6-month transthoracic echocardiography. At postoperative echocardiographic assessment, mean pressure gradient for the 25- through 29-mm size group was 10.2 +/- 7.1 mm Hg, and peak pressure gradient was 18.1 +/- 12.3 mm Hg. CONCLUSION: Our initial experience combined a well-established supra annular implantation technique with the Pericarbon Freedom stentless bioprosthesis, a latest-generation pericardial stentless valve. The combination showed excellent results in terms of safety and reliability, although this technique required adequate experience. Clinical outcomes are similar to those obtained with other techniques, with satisfactory hemodynamic performance. PMID- 16256778 TI - Risk factors for and economic implications of prolonged ventilation after cardiac surgery. AB - OBJECTIVE: The study's objective was to identify predictors of prolonged ventilation and assess clinical and cost implications in patients undergoing cardiac surgery. METHODS: Patients undergoing cardiac surgery were classified as (1) ventilated less than 96 hours or (2) ventilated 96 hours or more. Multivariate modeling was used to identify predictors of prolonged ventilation and to ascertain the impact of prolonged ventilation on in-hospital mortality and bed occupancy costs and 5-year survival. RESULTS: A total of 7553 patients were studied; 197 (2.6%) had prolonged ventilation. Median ventilation times were 8 and 192 hours, and in-hospital mortality was 1.0% and 22.2% in the control and prolonged ventilation groups, respectively (P < .001). In-hospital mortality remained higher in the prolonged ventilation group after adjustment and when comparing propensity-matched patients (odds ratio 8.06; 95% confidence interval [CI] 4.27-15.2; P < .001 for propensity-matched groups). Independent predictors of prolonged ventilation were as follows: older age, New York Heart Association class, ejection fraction less than 50%, creatinine greater than 200 micromol/L, multiple valve replacements, aortic procedures, operative priority, reoperation for bleeding, inotropes, and preoperative intra-aortic balloon pump. Five-year survival was lower in the prolonged ventilation group (56.1% [95% CI 46.6%-64.6%] vs 88.8% [95% CI 87.9%-89.6%]) also after adjustment for imbalances and when comparing propensity-matched patients (hazard ratio 2.39; 95% CI 1.75-3.27; P < .001 for propensity-matched groups). Mean bed occupancy costs were 14,286 dollars (95% CI 12,731 dollars-15,690 dollars) and 2761 dollars (95% CI 2705 dollars-2814 dollars) in the prolonged ventilation and control groups, respectively (P < .001). CONCLUSION: Prolonged ventilation is associated with high in-hospital mortality and costs, and poor 5-year survival. Identified predictors of prolonged ventilation might help to optimize the clinical management of these patients. PMID- 16256780 TI - Aorta-right atrial tunnel: clinical presentation, diagnostic criteria, and surgical options. AB - OBJECTIVE: Aorta-right atrial tunnel is a rare and distinct congenital anomaly. To the spectrum of aorta-right atrial communications, we want to add this separate entity of aorta-right atrial tunnel by presenting the clinical features, morphologic aspects, diagnostic criteria, surgical techniques, and outcome in various clinical settings for better understanding of this developmentally intriguing, clinically complex, and therapeutically challenging disorder. METHODS: From June 1994 through October 2004, 9 patients were treated for aorta right atrial tunnel at our institution. Ages ranged from 9 to 45 years. There were 5 male subjects and 4 female subjects. Morphologically, 2 types of tunnels were identified in relationship to the ascending aorta: the anterior type in 3 patients and the posterior type in 6 patients. In all patients the diagnosis was established by means of 2-dimensional echocardiography and transesophageal echocardiography and confirmed by means of angiography. Eight patients were treated surgically, and 1 patient was treated with coil embolization. Postoperative echocardiograms obtained for all patients before discharge confirmed complete obliteration of the tunnel. RESULTS: One patient died perioperatively, and the other 8 patients were discharged in stable condition. During follow-up at 3 months, 6 months, and 1 year, all patients were in New York Heart Association class I, and echocardiography showed no residual shunts. CONCLUSIONS: In patients with aorta-right atrial tunnel, 2-dimensional echocardiography and transesophageal echocardiography are enough to establish clinical diagnosis, but ascending aortography is necessary to differentiate from more common clinical conditions, like ruptured sinus of Valsalva aneurysm and coronary cameral fistula. The rarity of this condition is established by the fact that during the same period of time, we have treated in our institution 66 cases of ruptured sinus of Valsalva aneurysm, which is the most common aorta-right atrial communication. Treatment options are simple ligation or ligation with implantation of coronary ostium or coil embolization. The location of the coronary ostium dictates technical details. Follow-up reveals excellent functional recovery. PMID- 16256779 TI - Increasing duration of deep hypothermic circulatory arrest is associated with an increased incidence of postoperative electroencephalographic seizures. AB - OBJECTIVE: Electroencephalographic seizures have been shown to occur in 5% to 20% of neonates and infants after biventricular repair of a variety of cardiac defects. Occurrence of a seizure is a predictor of adverse long-term neurodevelopmental sequelae. The contemporary incidence of postoperative seizures after repair of cardiac defects such as hypoplastic left heart syndrome and other forms of single ventricle is not known. METHODS: A prospective study of 178 patients less than 6 months of age undergoing cardiopulmonary bypass with or without deep hypothermic circulatory arrest (DHCA) was conducted at a single institution from September 2001 through March 2003 to identify postoperative seizures assessed by 48-hour continuous video electroencephalographic monitoring. RESULTS: Cardiac defects included transposition of the great arteries with or without a ventricular septal defect (n = 12), ventricular septal defect with or without coarctation (n = 28), tetralogy of Fallot (n = 24), hypoplastic left heart syndrome or variant (n = 60), other functional single ventricle (n = 14), and other defects suitable for biventricular repair (n = 40). Median age at the time of the operation was 7 days (range, 1-188 days) and was 30 days or less in 110 (62%) patients. DHCA was used in 117 (66%) patients, with multiple episodes in 9 patients. Median total duration of DHCA was 40 minutes (range, 1-90 minutes). Electroencephalographic seizures were identified in 20 (11.2%) patients. Seizures occurred in 15 (14%) of 110 neonates and 5 (7%) of 68 older infants. Seizures occurred in 1 (4%) of 24 patients with tetralogy of Fallot, 1 (8%) of 12 with transposition of the great arteries, and 11 (18%) of 60 with hypoplastic left heart syndrome or variant. By stepwise logistic regression analysis, once increasing duration of total DHCA (P = .001) was considered, no other variable improved prediction of occurrence of a seizure. Patients with DHCA duration of more than 40 minutes had an increased incidence of seizures (14/58 [24.1%]) compared with those with a DHCA duration of 40 minutes or less (4/59 [6.8%], P = .04). The incidence of seizures for patients with a DHCA duration of 40 minutes or less was not significantly different from those in whom DHCA was not used (2/61 [3.3%], P = .38). CONCLUSIONS: In the current era, continuous electroencephalographic monitoring demonstrates early postoperative seizures in 11.2% of a heterogeneous cohort of neonates and infants with complex congenital heart defects. Increasing duration of DHCA was identified as a predictor of seizures. However, the incidence of seizures in children with limited duration of DHCA was similar to that in infants undergoing continuous cardiopulmonary bypass alone. PMID- 16256781 TI - Laryngopharyngeal dysfunction after the Norwood procedure. AB - OBJECTIVE: We sought to evaluate the incidence and significance of recurrent laryngeal nerve and swallowing dysfunction after a Norwood procedure compared with that after biventricular aortic arch reconstruction. METHODS: From April 2003 through December 2004, 36 neonates underwent a Norwood procedure; 33 of 36 had postoperative fiberoptic laryngoscopy and modified barium swallow. Study results were used to guide the transition from nasogastric tube to oral feeding and placement of gastrostomy tubes. During the same time period, 18 neonates underwent aortic arch reconstruction as part of a biventricular repair. RESULTS: After a Norwood procedure, laryngoscopy showed left true vocal fold (cord) paralysis in 3 (9%) of 33 patients. The results of a modified barium swallow were abnormal in 16 (48%) of 33 patients, with aspiration in 8 (24%) of 33 patients. Of the 3 patients with vocal fold paralysis, 2 had a normal modified barium swallow result, and 1 had aspiration. Gastrostomy tubes were placed in 6 (18%) of 33 patients, all with an abnormal modified barium swallow result. Hospital stay was longer in patients with an abnormal modified barium swallow result: 34 +/- 13 versus 22 +/- 7 days (P < .01). After biventricular repair with aortic arch reconstruction, left true vocal fold paralysis occurred in 4 (25%) of 16 patients; results of a modified barium swallow were abnormal in 10 (59%) of 17 patients, with aspiration in 6 (35%) of 17 patients (all nonsignificant vs patients undergoing the Norwood procedure). Follow-up laryngoscopy in 4 patients with vocal fold paralysis showed no change in 3 of 4 patients and improvement in 1 patient. Follow-up modified barium swallow showed resolution of aspiration in 11 (85%) of 13 patients. Hospital survival was 32 (89%) of 36 patients for the Norwood procedure and 18 (100%) of 18 patients for biventricular repair. There has been 1 sudden death before second-stage palliation. CONCLUSIONS: After a Norwood procedure, swallowing dysfunction occurs in 48% of patients, with aspiration in 24%, and results in increased length of hospital stay. Left recurrent laryngeal nerve injury, seen in 9% of patients, is an uncommon cause of swallowing dysfunction. Postoperative aspiration generally resolves over time, whereas vocal fold paralysis does not. Systematic evaluation of swallowing function allows appropriate tailoring of feeding regimens and might contribute to decreased hospital and interstage mortality. PMID- 16256782 TI - Clinical outcomes in patients with chronic congestive heart failure who undergo left ventricular assist device implantation. AB - OBJECTIVE: The use of left ventricular assist devices as a bridge to transplantation for patients with chronic congestive heart failure is well accepted. However, few studies have examined outcomes solely for these patients. This study details one center's left ventricular assist device experience with this population. METHODS: Two hundred one patients received HeartMate left ventricular assist devices (Thoratec Corp, Pleasanton, Calif) from January 1, 1996, to April 30, 2004. Of these, 119 (59.2%) had chronic congestive heart failure (diagnosis >6 months) as the primary indication. Outcome parameters included early mortality after left ventricular assist device placement (<30 days), bridge-to-transplantation rate, and posttransplantation survival. Variables examined included patient demographic data; preoperative pacemaker, internal defibrillator, and balloon pump use; and preoperative laboratory values. RESULTS: Advanced age, female sex, and diabetes were independent predictors of early death (P = .048, odds ratio 1.879 per 10 years of age, 95% confidence interval 1.005-3.515; P = .002, odds ratio 10.029, 95% confidence interval 2.256 44.583; P = .040, odds ratio 3.974, 95% confidence interval 1.063-14.861). Advanced age, female sex, and low preoperative albumin were independent predictors of poor bridge-to-transplantation rate (P = .029, odds ratio 0.135 per 10 years of age, 95% confidence interval 0.022-0.819; P = .002, odds ratio 0.013, 95% confidence interval 0.001-0.197; P = .023, odds ratio 19.178 per 1 g/dL albumin, 95% confidence interval 1.504-244.598). There were no independent predictors of poor posttransplantation survival and prolonged intensive care unit stay. Overall bridge-to-transplantation rate was 81.5%. The 1-, 3-, 5-, and 7 year posttransplantation survivals were 88.4%, 84.5%, 78.4%, and 76.0%. CONCLUSION: Among patients with chronic congestive heart failure, advanced age, female sex, diabetes, and low preoperative albumin predict poor clinical course. Careful risk stratification and comprehensive evaluation by care providers should be performed for candidates who are female, are elderly, and have diabetes, and preoperative nutritional optimization should be encouraged to enhance patient outcomes. PMID- 16256783 TI - Cell transplantation preserves cardiac function after infarction by infarct stabilization: augmentation by stem cell factor. AB - OBJECTIVE: We hypothesized that implantation of adult mesenchymal stem cells after acute myocardial infarction mobilizes bone marrow precursor cells by activating the stem cell factor pathway, and that overdriving this pathway would enhance the beneficial effects of cell transplantation. METHODS: After coronary ligation, medium, mesenchymal stem cells, or stem cell factor-overproducing mesenchymal stem cells were injected into the anterior left ventricle. Cells from beta-galactosidase transgenic mice enabled tracking of injected cells. The global and local impact of the cells was evaluated by measuring cytokine levels, endothelial progenitor cells, and myocardial angiogenesis, and by addressing cardiomyogenesis with confocal microscopy. The impact on cardiac function was evaluated by pressure-volume loops. Ventricular morphometrics were measured after in situ perfusion-fixation of the hearts at physiologic pressures. RESULTS: Implantation of mesenchymal stem cells increased myocardial stem cell factor levels 2.0-fold, endothelial progenitor cell mobilization 2.7-fold, and myocardial angiogenesis 2.3-fold (P < .05), but did not induce mitogenesis in host cardiomyocytes or give rise to beta-galactosidase-expressing cardiomyocytes. Cell-transplanted groups had improved indices of cardiac function, including preload recruitable stroke work and end-systolic elastance (P < .001). Cell transplantation resulted in 2.0-fold smaller ventricular volumes (P = .001) and 2.0-fold reduced infarct scar area (P = .056), but had no effect on the volume of spared myocardium. Stem cell factor overproduction imparted greater functional benefit without inducing detectable histologic cardiomyocyte regeneration. CONCLUSION: Mesenchymal stem cell implantation after myocardial infarction facilitates functional cardiac regeneration without myocyte regeneration through augmentation of endogenous infarct repair, which is enhanced by stem cell factor. PMID- 16256784 TI - Attempted control of hyperglycemia during cardiopulmonary bypass fails to improve neurologic or neurobehavioral outcomes in patients without diabetes mellitus undergoing coronary artery bypass grafting. AB - OBJECTIVE: Hyperglycemia worsens outcomes in critical illness. This randomized, double-blind, placebo-controlled clinical trial tested whether insulin treatment of hyperglycemia during cardiopulmonary bypass would reduce neurologic, neuro ophthalmologic, and neurobehavioral outcomes after coronary artery bypass grafting. METHODS: Three hundred eighty-one nondiabetic patients undergoing isolated coronary artery bypass grafting were given infusions of insulin or placebo when their blood glucose concentration exceeded 100 mg/dL during cardiopulmonary bypass. The primary outcome measure was the combined incidence of new neurologic, neuro-ophthalmologic, or neurobehavioral deficits or neurologic death observed at 4 to 8 days postoperatively. This same measure was assessed secondarily at 6 weeks and 6 months. Length of hospital stay was also compared as a secondary assessment. RESULTS: The 2 groups were well matched at baseline. The insulin-treated group had significantly lower blood glucose concentrations during bypass. Sixty-six percent of subjects in the insulin-treated group and 67% of subjects in the control group demonstrated a new or worsening neurologic, neuro ophthalmologic, or neurobehavioral deficit or neurologic death at the 4- to 8-day assessment. Outcomes were also similar in the 2 groups at 6 weeks (37% and 39% incidence, respectively) and 6 months (30% and 25%, respectively). Median lengths of stay were 7 and 6 days, respectively, in the treatment and control groups. None of these outcome differences was statistically significant. CONCLUSION: Attempted control of hyperglycemia during cardiopulmonary bypass had no significant effect on the combined incidence of neurologic, neuro-ophthalmologic, or neurobehavioral deficits or neurologic death and failed to shorten the length of hospital stay. These results do not contradict those of other studies showing that aggressive control of hyperglycemia in the postoperative period will improve outcome. PMID- 16256785 TI - The remote ischemic preconditioning stimulus modifies gene expression in mouse myocardium. AB - BACKGROUND: We have recently demonstrated that remote ischemic preconditioning reduces ischemia-reperfusion injury in animal models. The mechanisms by which the remote ischemic preconditioning stimulus exerts its effect remain to be fully defined, and its effect on myocardial gene expression is unknown. We tested the hypothesis that remote ischemic preconditioning modifies myocardial gene expression immediately after the remote ischemic preconditioning stimulus (early phase) and 24 hours later (late phase). METHODS: Twenty male (C57BL/6) 10- to 12 week-old mice were randomized into 4 groups: group 1 (control, early phase; n = 5), group 2 (remote ischemic preconditioning, early phase; n = 5), group 3 (control, late phase; n = 5), and group 4 (remote ischemic preconditioning, late phase; n = 5). Groups 2 and 4 underwent remote ischemic preconditioning induced by 6 cycles of 4 minutes of occlusion and 4 minutes of reperfusion of the femoral artery. Groups 1 and 2 were killed 15 minutes after completion of sham procedure or remote ischemic preconditioning, and the hearts were removed and frozen in liquid nitrogen. Groups 3 and 4 were killed 24 hours after remote ischemic preconditioning, and the hearts were harvested in the same fashion. Gene expression was assessed by using the Affymetrix MG-430A chip (Affymetrix, Santa Clara, Calif). RESULTS: Data filtering (P < .05, analysis of variance) and hierarchic 2-way clustering identified significant differences in gene expression among the 4 groups. Genes involved in protection against oxidative stress (eg, Hadhsc, Prdx4, and Fabp4) and cytoprotection (Hsp73) were upregulated, whereas many proinflammatory genes (eg, Egr-1 and Dusp 1 and 6) were suppressed. CONCLUSION: A simple remote ischemic preconditioning stimulus modifies myocardial gene expression by upregulating cardioprotective genes and suppressing genes potentially involved in the pathogenesis of ischemia-reperfusion injury. PMID- 16256786 TI - Repair of impaired myocardium by means of implantation of engineered autologous myoblast sheets. AB - OBJECTIVES: Autologous skeletal myoblast cell transplantation by means of the injection method is subject to the loss of intercellular communication, extracellular matrix, and cell numbers. We hypothesize that the implantation of skeletal myoblast cell sheets might be more advantageous in repairing the impaired heart by providing uniform and stable cell delivery with less cell loss and without disrupting the cell-cell microenvironment. METHODS: Left anterior descending coronary artery-ligated Lewis rat hearts (2 weeks, total n = 173) received 1 x 10(7) autologous skeletal myoblasts by means of cell transplantation either through myoblast injection or implantation of 2 monolayer-constructed myoblast sheets (5 x 10(6) cells per sheet) or through medium injection. Myoblast sheets were constructed with temperature-responsive, polymer-grafted cell-culture dishes, which release the confluent cells from the dish surface at less than 20 degrees C. RESULTS: Echocardiographic results indicated higher improvement of cardiac performance in the myoblast sheet group than among the other groups until 8 weeks after cell transplantation. Histologic comparison revealed greater cellularity and abundant widespread neocapillaries within the noticeable uniform thickened wall in myoblast sheet group hearts only. Fibrosis was substantially reduced with skeletal myoblast sheet implantation compared with skeletal myoblast cell injection. Obviously higher numbers of hematopoietic stem cells (c-kit, stem cell antigen 1, and CD34) were observed in the myoblast sheet group infarct heart region. Reverse transcription-polymerase chain reaction results showed expression of stromal-derived factor 1, hepatocyte growth factor, and vascular endothelial growth factor as follows: myoblast sheets > myoblast injection > control. CONCLUSIONS: Myoblast sheets repaired the impaired myocardium, reduced fibrosis, and prevented remodeling in association with recruitment of hematopoietic stem cells through the release of stromal-derived factor 1 and other growth factors. Our experiment indicates a therapy for patients with severe heart failure. PMID- 16256787 TI - Circulating big endothelin-1: an active role in pulmonary thromboendarterectomy? AB - BACKGROUND: Pulmonary thromboendarterectomy is an effective treatment for patients with chronic thromboembolic pulmonary hypertension. The early postoperative course may be associated with pulmonary vasoconstriction and profound systemic vasodilation. We investigated the potential involvement of endothelins in these hemodynamic alterations. METHODS: Seventeen patients with chronic thromboembolic pulmonary hypertension (pulmonary vascular resistance, 1015 +/- 402 dyne x s x cm(-5) [mean +/- SD]) underwent pulmonary thromboendarterectomy with cardiopulmonary bypass and deep hypothermic circulatory arrest. Peripheral arterial blood samples were drawn before sternotomy, during cardiopulmonary bypass before and after deep hypothermic circulatory arrest, and 0, 8, 16, and 24 hours after surgery and were analyzed for big endothelin-1. The patients were divided into 2 groups according to whether their preoperative big endothelin-1 plasma level was above or below the cutoff point of 2.1 pg/mL, as determined by receiver operating characteristic curve analysis (group A, big endothelin-1 <2.1 pg/mL, n = 8; group B, big endothelin-1 > or =2.1 pg/mL, n = 9). RESULTS: Patients in group B, with higher preoperative big endothelin-1 levels (3.2 +/- 1.0 pg/mL vs 1.5 +/- 0.4 pg/mL; P < .001), were poorer operative candidates (preoperative mean pulmonary artery pressure, 51.3 +/- 7.1 mm Hg vs 43.6 +/- 6.2 mm Hg; P = .006) and had a poorer outcome (mean pulmonary artery pressure 24 hours after surgery, 32.6 +/- 9.5 mm Hg vs 21.8 +/- 6.2 mm Hg; P < .001). Positive correlations were found between preoperative big endothelin-1 levels and preoperative mean pulmonary artery pressure (r = 0.56; P = .02) as well as postoperative mean pulmonary artery pressure at 0 hours (r = 0.70; P = .002) and 24 hours (r = 0.63; P = .006) after surgery. Preoperative big endothelin-1 levels predicted outcome (postoperative mean pulmonary artery pressure at 24 hours after surgery) after pulmonary thromboendarterectomy (area under the receiver operating characteristic curve, 0.85). Peak big endothelin-1 levels also correlated with maximal vasopressor dosage (r = 0.65; P = .004). CONCLUSIONS: Preoperative big endothelin-1 levels seem to correlate with the hemodynamic alterations observed in pulmonary thromboendarterectomy and may be used to predict hemodynamic outcome after pulmonary thromboendarterectomy. PMID- 16256788 TI - Three-dimensional echo-guided beating heart surgery without cardiopulmonary bypass: atrial septal defect closure in a swine model. AB - OBJECTIVE: In this study, we tested 3 techniques of atrial septal defect closure under real-time 3-dimensional echocardiography guidance in a swine model. METHODS: The operations were conducted under the sole guidance of a modified real time 3-dimensional echocardiography guidance system with a x4 matrix transducer (Sonos 7500, Philips Medical Systems, Andover, Mass). Eighteen swine were anesthetized, and after median sternotomy, the echo probe was applied directly to the surface of the right atrium. To create an atrial septal defect, balloon atrial septostomy and atrial septal defect enlargement were performed. Subsequently, 3 different techniques of atrial septal defect closure were attempted: group I, direct suture closure; group II, closure of the atrial septal defect using the Amplatzer device (AGA Medical Corp, Golden Valley, Minn); and group III, patch closure of the atrial septal defect (n = 6 each). RESULTS: Real time 3-dimensional echocardiography guidance provided sufficient spatial resolution and a satisfactory frame rate to provide a "virtual surgeon's view" of the relevant anatomy during the entire procedure. All atrial septal defects were enlarged, and the mean final size was 8.5 +/- 1.8 mm. Atrial septal defect closure was successfully accomplished with all the 3 surgical techniques examined. In groups I and III, the needles (1-3 sutures) and staples (6-12 staples) penetrated the tissue and patch material consistently, whereas in group III, the Amplatzer atrial septal defect device was easily deployed. There was no incident device/staple embolization or air introduction. Neither intraoperative 2 dimensional color Doppler echocardiography nor postmortem macro-evaluation revealed any residual shunts. CONCLUSIONS: Beating heart atrial septal defect closure under real-time 3-dimensional echocardiographic guidance is feasible and, unlike catheter-based devices, applicable for any type of secundum atrial septal defect. PMID- 16256789 TI - In vitro tissue engineering of a cardiac graft using a degradable scaffold with an extracellular matrix-like topography. AB - OBJECTIVE: Cardiac tissue engineering has been proposed as a treatment to repair impaired hearts. Bioengineered cardiac grafts are created by combining autologous cell transplantation with a degradable scaffold as a temporary extracellular matrix. Here we present a system for engineered myocardium combining cultured cardiomyocytes and a novel biodegradable scaffold with a unique extracellular matrix-like topography. METHODS: Cardiomyocytes were harvested from neonatal rats and cultured in vitro on biodegradable electrospun nanofibrous poly(epsilon caprolactone) meshes. Between days 5 and 7, the meshes were overlaid to construct 3-dimensional cardiac grafts. On day 14 of in vitro culture, the engineered cardiac grafts were analyzed by means of histology, immunohistochemistry, and scanning electron microscopy. RESULTS: The cultured cardiomyocytes attached well to the meshes, and strong beating was observed throughout the experimental period. The average fiber diameter of the scaffold is about 250 nm, well below the size of an individual cardiomyocyte. Hence the number of cell-cell contacts is maximized. Constructs with up to 5 layers could be formed without any incidence of core ischemia. The individual layers adhered intimately. Morphologic and electrical communication between the layers was established, as verified by means of histology and immunohistochemistry. Synchronized beating was also observed. CONCLUSIONS: This report demonstrates the formation of thick cardiac grafts in vitro and the versatility of biodegradable electrospun meshes for cardiac tissue engineering. It is envisioned that cardiac grafts with clinically relevant dimensions can be created by using this approach and combining it with new technologies to induce vascularization. PMID- 16256790 TI - Efficient myocyte gene delivery with complete cardiac surgical isolation in situ. AB - BACKGROUND: Previously, we used cardiopulmonary bypass with incomplete cardiac isolation and antegrade administration of vector for global cardiac gene delivery. Here we present a translatable cardiac surgical procedure that allows for complete surgical isolation of the heart in situ with retrograde (through the coronary venous circulation) administration of both vector and endothelial permeabilizing agents to increase myocyte transduction efficiency. METHODS: In 6 adult dogs the heart was completely isolated with tourniquets placed around both vena cavae and cannulas and all pulmonary veins. On cardiopulmonary bypass, the aorta and pulmonary artery were crossclamped, and the heart was isolated. Crystalloid cardioplegia at 4 degrees C containing 10(13) particles of adenovirus encoding LacZ and 15 microg of vascular endothelial growth factor was infused retrograde into the coronary sinus and recirculated for a total of 30 minutes. The dogs were then weaned from cardiopulmonary bypass and allowed to recover. With a catheter, 3 control dogs underwent retrograde infusion of the same cocktail without cardiac isolation or cardiopulmonary bypass. RESULTS: Beta galactosidase activities in the cardiopulmonary bypass group were several orders of magnitude higher in both the right and left ventricles when compared with those in the control group (P < .05). X-gal staining from the cardiopulmonary bypass group showed unequivocal evidence of myocyte gene expression globally in a significant proportion of cardiac myocytes. No myocyte gene expression was observed in the control group. CONCLUSION: A novel cardiac surgical technique has been developed. This approach with cardiac isolation and retrograde delivery of vector through the coronary sinus results in efficient myocyte transduction in an adult large animal in vivo. PMID- 16256791 TI - Promoter methylation of the hMLH1 gene and protein expression of human mutL homolog 1 and human mutS homolog 2 in resected esophageal squamous cell carcinoma. AB - OBJECTIVE: Aberrant expression of mismatch repair genes, such as human mutL homolog 1 (hMLH1) and human mutS homolog 2 (hMSH2), are common in some human cancers, and promoter methylation is believed to inactivate expression of hMLH1. We investigated whether promoter methylation is involved in loss of hMLH1 protein and whether aberrant expression of hMLH1 and hMSH2 protein is related to prognosis after resection for esophageal squamous cell cancer. METHODS: We analyzed promoter methylation of hMLH1 using methylation-specific polymerase chain reaction and hMLH1 and hMSH2 protein by using immunohistochemistry in 60 resected tumor specimens. The Pearson chi2 test was used to compare expression of hMLH1 and hMSH2 protein among patients with different clinicopathologic parameters. Concordance analysis was performed between hMLH1 methylation and its protein expression. RESULTS: Loss of hMLH1 and hMSH2 protein was found in 43 (72%) and 39 (65%, P = .06) of 60 resected specimens, respectively. hMLH1 protein correlated well with tumor staging (P < .0001), depth of tumor invasion (P = .008), and nodal involvement (P < .0001) but not with distant metastasis, whereas hMSH2 did not show correlation with any of these parameters. A concordance rate of 83.3% was present between expression of hMLH1 protein and its promoter methylation (P < .001). CONCLUSIONS: Aberrant expression of hMLH1 and hMSH2 protein is frequently associated with the presence of esophageal squamous cell carcinoma, and expression of hMLH1 protein is a better prognostic predictor than is expression of hMSH2 protein. Promoter methylation is one of the mechanisms responsible for loss of hMLH1 protein in esophageal squamous cell cancer. PMID- 16256792 TI - Prognostic implication of aberrant promoter hypermethylation of CpG islands in adenocarcinoma of the lung. AB - OBJECTIVES: DNA hypermethylation in promoter regions has been studied for various types of cancer. However, there is no clear evidence that shows whether methylation status can predict long-term survival in patients with lung cancer. METHODS: We collected tissues from 72 patients with lung adenocarcinomas. The cancer and normal lung tissues were tested for DNA hypermethylation by using methylation-specific polymerase chain reaction. The genes investigated were p16INK4alpha(p16), retinoic acid receptor beta-promoter (RARbetaP2), death associated protein kinase (DAPK), O6-methylguanine-DNA-methyltransferase (MGMT), and glutathione-S-transferase P1 (GSTP1). The status of the DNA methylation was analyzed, and we focused on long-term outcomes, as well as other clinical variables. RESULTS: DNA hypermethylation was observed in 83% for p16, 63% for RARbetaP2, 32% for DAPK, 17% for MGMT, and 46% for GSTP1 from the cancer tissue. From normal lung tissue, the results of methylation were positive in 75% for p16, 24% for RARbetaP2, 10% for DAPK, 6% for MGMT, and 33% for GSTP1. During the mean follow-up period of 18 +/- 11 months (1-40 months), 25 (35%) patients experienced recurrence, and 13 died. In multivariable analysis, old age (>60 years, P = .007), male sex (P = .004), unmethylation of DAPK from cancer tissue (P = .045), and hypermethylation of RARbetaP2 from normal tissue (P = .000) were risk factors for poor survival. Pathologic stage (P = .023), unmethylation of DAPK from normal tissue (P = .043), and hypermethylation of RARbetaP2 from normal tissue (P = .030) were risk factors for disease-free survival. CONCLUSIONS: DNA methylation status of CpG islands seems to be a useful predictor of long-term outcome for adenocarcinoma of the lung. However, because the predictive power is still low, further studies, including those with multiple genes, are necessary to increase its usefulness in the clinical setting. PMID- 16256793 TI - Surgical results for bronchiectasis based on hemodynamic (functional and morphologic) classification. AB - OBJECTIVE: This study was a prospective evaluation of surgical indications and outcomes for unilateral and bilateral bronchiectasis according to hemodynamic (functional and morphologic) classification. METHODS: Between January 1998 and January 2004, the morphologic features (cystic versus cylindric) by chest computed tomography and the hemodynamic features (perfused versus nonperfused) by lung ventilation/perfusion scan were determined in 66 patients with bronchiectasis (53 unilateral and 13 bilateral). The indication for surgical resection in both groups was the presence of localized areas of cystic, nonperfused bronchiectasis. RESULTS: In the unilateral bronchiectasis group, there were 28 female and 25 male patients with an average age of 37.5 +/- 3.8 years (range 6-40 years). Pneumonectomy was performed in 10 cases (8 left and 2 right), and lobectomy or bilobectomy was performed in 43. In the bilateral group, there were 7 male and 6 female patients with an average age of 42 +/- 5.4 years (range 9-55 years). Pneumonectomy was performed in 2 cases, lobectomy in 5, and bilateral staged lobectomy in 6. There was 1 postoperative death (1.5%), and morbidity was 18% (12 patients). Four patients required reexploration for bleeding, 4 had prolonged air leak develop, 3 acquired pulmonary infections, and 1 had localized empyema develop. During a mean follow-up of 52 months (range 24 82 months), 48 patients were considered cured (73%) and 17 had symptomatic improvement (26%). Pseudomonas infection and underlying chronic obstructive airway disease were poor prognostic factors (P < .05). CONCLUSION: The hemodynamic (functional and morphologic) classification provides an accurate functional classification for bronchiectasis. Its application in determining the indications and extent of surgical resection is superior to morphologic classification alone. Curative resection can be achieved in both unilateral and bilateral bronchiectasis with acceptable morbidity. PMID- 16256794 TI - Long-segment, supercharged, pedicled jejunal flap for total esophageal reconstruction. AB - OBJECTIVE: Many patients with cancer have limited esophageal reconstruction options when the stomach is unavailable as a replacement conduit or when long segment discontinuity exists. Jejunum has been used as an alternative conduit, both as a pedicled or free flap interposition; however, reports of this are usually limited to short-segment repairs. Microvascular augmentation of a pedicled jejunal flap allows creation of a longer conduit, making it possible to replace the entire esophagus with jejunum. Few reports describe this technique in patients with cancer. We report our initial experience with "supercharged" pedicled jejunum as an alternative conduit for total esophageal reconstruction. METHODS: Review of a prospectively collected departmental database was performed to identify those patients who underwent total esophageal reconstruction with supercharged pedicled jejunum. Data regarding their perioperative course and postoperative function were gathered from the prospectively collected clinical data, review of hospital records, and patient interviews. RESULTS: Total esophageal reconstruction with supercharged pedicled jejunum was attempted in 26 patients (age range, 37-74 years) between March 2000 and April 2004. Twenty-four of 26 patients were ultimately discharged with an intact supercharged pedicled jejunum flap, for an overall success rate of 92.3%. One patient experienced intraoperative flap loss caused by technical difficulties harvesting the flap and never had the flap interposed. One other flap loss occurred in the early postoperative period in a patient who had multisystem organ failure after a prolonged reconstruction. Cervical anastomotic leaks occurred in 19.2% (5/26) of the patients. Two midconduit leaks occurred that were suspicious for iatrogenic perforation from nasogastric tube placement; one required reoperation. One additional early reoperation was performed for cecal ischemia. There were no mortalities. Functional results were available in 95.4% (21/22) of the patients receiving supercharged pedicled jejunum who survived at least 6 months after reconstruction. At the time of follow-up, 95% (20/21) of the patients were tolerating regular diet, and 76.2% (16/21) did not require any supplemental alimentation. Ninety-five percent (20/21) of the patients were free from reflux symptoms, and 80.9% (17/21) had no dumping symptoms. Only 1 patient required dilation of a midconduit stricture. One patient required late reoperation for conduit redundancy. CONCLUSIONS: Supercharged pedicled jejunum is a suitable alternative conduit for total esophageal replacement in patients with cancer with otherwise limited reconstructive options. Functional outcomes are excellent, despite the severity of disease and technical challenges in this patient population. PMID- 16256795 TI - Extensive circumferential endoscopic mucosal resection with a new rigid esophagoscope: an animal study. AB - OBJECTIVE: Current endoscopic mucosal resection techniques are suboptimal for large circumferential mucosal resections intended to treat Barrett mucosa with multicentric foci of high-grade intraepithelial neoplasia or early adenocarcinomas. A rigid modified endoscope was developed for extended endoscopic mucosal resection in the esophagus. This pilot animal study investigated the feasibility of circumferential endoscopic mucosal resections of different lengths in the sheep esophagus. METHODS: Circumferential esophageal endoscopic mucosal resections of 2.2 cm (n = 6), 3.3 cm (n = 6), 4.4 cm (n = 7), and 5.5 cm (n = 5) were performed in 24 sheep. Circumferential resections consisted of two opposite hemicircumferential mucosectomies. Animals were followed up with endoscopic examinations at 1 week and then monthly to 6 months, or until complete re epithelialization without stenosis eventually ensued. Strictures were treated with a single or repeated dilatations with Savary bougies. RESULTS: Circumferential resections 2.2 to 5.5 cm in length were successfully performed in 23 of 24 of the animals. One perforation occurred directly after mucosectomy. Accurate depth of resection through the submucosa was obtained in 85% of the specimens. Cicatricial stenosis was controlled in 95% by bougienage. CONCLUSION: Endoscopic mucosal resection performed with the rigid esophagoscope allowed extensive circumferential resections in a single session. There is thus potential to eradicate complete Barrett esophagus with high-grade intraepithelial neoplasia or early adenocarcinomas. PMID- 16256796 TI - Dimethyl celecoxib as a novel non-cyclooxygenase 2 therapy in the treatment of non-small cell lung cancer. AB - OBJECTIVES: The cyclooxygenase 2 enzyme has become a therapeutic target in cancer treatment. Cyclooxygenase 2 blockade with selective inhibitors increases apoptosis and decreases the metastatic potential of lung cancer cells. Some of the antitumor effects of these inhibitors may occur through both cyclooxygenase 2 dependent and independent pathways. Our goal was to investigate these pathways using celecoxib (selective cyclooxygenase 2 inhibitor) and 2,5-dimethyl celecoxib, a structural analog modified to eliminate cyclooxygenase 2 inhibitory activity, while potentially maintaining antineoplastic properties. METHODS: 2,5 dimethyl celecoxib was synthesized in the Department of Chemistry at the University of Southern California. With the use of non-small cell lung cancer cells (A549), prostaglandin E2 production was quantified by enzyme-linked immunosorbent assay to assess cyclooxygenase 2 activity. Cell proliferation was assessed by 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4 sulfophenyl)-2H-tetrazolium, inner salt assay. Cell migration was performed using transwell inserts that were matrigel coated for invasion experiments. Gelatin zymography was used to assess matrix-metalloproteinase activity. RESULTS: 2,5 dimethyl celecoxib did not inhibit interleukin-1beta-stimulated prostaglandin E2 production, whereas celecoxib did even at low doses. Both celecoxib and 2,5 dimethyl celecoxib decreased tumor cell viability and proliferation with IC50 for celecoxib and 2,5-dimethyl celecoxib of 73 and 53 micromol/L, respectively. Both drugs were also potent inducers of apoptosis, and both inhibited tumor cell migration and invasion. This was associated with down-regulation of matrix metalloproteinase activity. CONCLUSIONS: 2,5-dimethyl celecoxib is a structural analog of celecoxib that lacks cyclooxygenase 2 inhibitory activity but exhibits significant antineoplastic properties comparable to celecoxib. This suggests that the antineoplastic activities of celecoxib are, at least in part, cyclooxygenase independent and that therapeutic strategies can be developed without the side effects of global cyclooxygenase 2 blockade. PMID- 16256797 TI - Predictors of recurrence in thymic tumors: importance of invasion, World Health Organization histology, and size. AB - OBJECTIVE: This study sought to define predictors of recurrence after resection of thymic tumors. METHODS: A single-institution retrospective study was performed of 179 patients who underwent resection of a thymic tumor from 1972 through 2003. RESULTS: Resection was complete in 90% (161/179) of patients. After a median follow-up of 115 months, the recurrence rate was 11% (20/179), the tumor-related death rate was 7.8% (14/179), and the overall death rate was 36.3% (65/179). Tumor recurrence correlated with advanced stage and histology (P < .0001). The difference in recurrence between Masaoka stage I (0) and II (1.7% [1/59]) was insignificant. Recurrence rates correlated with World Health Organization tumor type: A and AB, 0%; B1 and B2, 8% (4/51); B3, 27% (14/51); and C, 50% (2/4; P < .0001). Tumor size separation into quintiles demonstrated a step-up of recurrence at 8 cm (<8 cm, 1.8% [2/113]; > or =8 cm, 28% [18/64]; P < .003). Multivariate Cox modeling demonstrated that Masaoka stage (odds ratio, 5.70; P < .001), World Health Organization histology (odds ratio, 5.77; P = .003), and size (odds ratio, 1.16; P = .001) were independent predictors of recurrence. CONCLUSION: The Masaoka staging system could be collapsed to 3 degrees of invasion by combining stages I and II. The World Health Organization histologic type can be simplified for clinical use into A (A, AB), early B (B1, B2), advanced B (B3), and C tumors. Size of 8 cm or larger is an independent risk factor, even when patients with Masaoka stage III tumors are considered alone, and might identify candidates for preoperative therapy. PMID- 16256798 TI - Inhibition of phosphatidylinositol 3-kinase/Akt and histone deacetylase activity induces apoptosis in non-small cell lung cancer in vitro and in vivo. AB - OBJECTIVE: Resistance to histone deacetylase inhibitors in non-small cell lung cancer is mediated in part through activation of nuclear factor-kappaB through a phosphatidylinositol 3-kinase/Akt-dependent pathway. We hypothesize that inhibition of phosphatidylinositol 3-kinase/Akt will sensitize non-small cell lung cancer cells to histone deacetylase inhibitor-induced apoptosis. METHODS: Tumorigenic non-small cell lung cancer cell lines H157, H358, H460, and A549 were treated with nothing, the histone deacetylase inhibitor butyrate, the phosphatidylinositol 3-kinase/Akt inhibitor LY294002, or both compounds. Nuclear factor-kappaB activity was assessed by reporter gene assays and reverse transcriptase-polymerase chain reaction of the nuclear factor-kappaB dependent genes cIAP-2, Bfl/A1, and MnSOD. Whole cell extracts were immunoblotted for phospho-Akt, Akt, and phospho-ser/thr-Akt substrate. Cell death and apoptosis were measured by crystal violet staining, caspase-3 activity, and DNA fragmentation. A549 non-small cell lung cancer xenografts were created in athymic nude mice, and tumor growth was assessed after treatments as noted above. Explanted tumors underwent terminal deoxynucleotide transferase-mediated dUTP nick-end labeling and Western blot analyses for apoptosis assessment and drug target validation, respectively. RESULTS: Butyrate activated nuclear factor kappaB-dependent transcription, and LY294002 abrogated this effect. Combined treatment induced more apoptosis and cell death in vitro compared with either drug alone as measured by caspase-3, DNA fragmentation, and clonogenic survival. Combined butyrate and LY294002 was tumoristatic in vivo, but all other xenografts grew. This decreased tumor growth correlated with more apoptosis in the xenografts treated with combined therapy. Tumor levels of phospho-Akt and acetylated histone H3 were decreased and increased, respectively, in xenografts treated with combined therapy. CONCLUSIONS: Combined histone deacetylase inhibitor and phosphatidylinositol 3-kinase/Akt pathway inhibition sensitized non small cell lung cancer xenografts to apoptosis. Further investigations of this combined therapy are warranted as new pharmacologic phosphatidylinositol 3 kinase/Akt pathway inhibitors are developed. PMID- 16256799 TI - Cell transplantation preserves matrix homeostasis: a novel paracrine mechanism. AB - OBJECTIVES: Cell transplantation prevents chamber dilatation, but the underlying molecular mechanisms remain undefined. Structural cardiac remodeling involves matrix degradation from an imbalance of matrix metalloproteinases (MMP) relative to endogenous tissue inhibitors of metalloproteinases (TIMP). We aimed to determine the capacity of cell transplantation to alter extracellular matrix in the failing heart and, in so doing, identify novel paracrine molecular mediators underlying the beneficial effects of cell transplantation on chamber dilatation. METHODS: Smooth muscle cells were transplanted to the dilating left ventricle of cardiomyopathic hamsters (CTX, n = 15) compared with age-matched media-injected cardiomyopathic (CON, n = 15) and normal hamsters (n = 7). After 5 weeks, left ventricular volume was measured by computerized planimetry. Fibrillar collagen was examined by confocal microscopy. Matrix homeostasis was quantified by measuring MMP/TIMP expression/activity relative to myocardial collagen synthesis (14C-proline uptake). RESULTS: Left ventricular dilatation was attenuated in CTX hearts (P = .02). CTX restored perimysial collagen fiber content and architecture to normal levels. TIMP-2 and TIMP-3 expression were enhanced in CTX (TIMP-2, 195% +/- 42% of CON, P = .02; TIMP-3, 118% +/- 3% of CON, P = .002), and correspondingly, gelatinase MMP-2 activity was reduced (P < .05). The TIMP:MMP ratio was increased in CTX hearts (TIMP-2 to MMP-2, 410% +/- 134% of CON, P = .04, and TIMP-3 to MMP-9, 205% +/- 47% of CON, P = .03), reflecting a reduced capacity for matrix degradation. Collagen synthesis was equivalent (CTX vs CON), suggesting that restored matrix architecture was a function of attenuated matrix degradation. CONCLUSIONS: These data provide the first evidence that cell transplantation limits ventricular dilatation in the failing heart through a paracrine-mediated mechanism that preserves extracellular matrix homeostasis. PMID- 16256800 TI - Hyperoxic ventilation exacerbates lung reperfusion injury. AB - OBJECTIVE: It is well known that hyperoxia can be potentially harmful to the ventilated patient, although little is known about the potential effects in the setting of lung reperfusion. We hypothesized that hyperoxic ventilation at the time of reperfusion could worsen the effects of lung reperfusion injury. METHODS: Using an ex vivo, blood perfused, isolated rabbit lung system, we evaluated the effects of hyperoxic (fraction of inspired oxygen = 100%, n = 10) versus normoxic (room air, n = 10) ventilation after 18 hours of cold ischemia. Lungs were ventilated and perfused for 2 hours. A control group was immediately perfused and ventilated with a fraction of inspired oxygen of 100%. RESULTS: Lung wet/dry ratios demonstrated lower tissue edema in the normoxic group compared with in the hyperoxic group (6.72 +/- 0.89 vs 7.62 +/- 1.14 [mean +/- standard error of the mean], P = .04). Lung ventilation was also significantly better in the normoxic group versus the hyperoxic group (PCO2 = 28.96 +/- 2.01 vs 36.68 +/- 3.20 mm Hg, P = .04). Conversely, lung oxygenation after 2 hours of reperfusion (normoxic group ventilated for the last 15 minutes on 100% fraction of inspired oxygen) was not significantly different between groups (PO2 = 590.2 +/- 50.1 vs 499.6 +/- 67.5 mm Hg, P = .25). CONCLUSIONS: Ventilating lungs with 100% fraction of inspired oxygen at the time of reperfusion could increase the risk of lung reperfusion injury at the time of transplantation. Thus the patient should be ventilated with as low a fraction of inspired oxygen as possible to achieve adequate oxygen saturations during this critical reperfusion period. PMID- 16256802 TI - Systolic anterior motion after mitral valve repair: an exceptional cause of late failure. PMID- 16256801 TI - Unexpected donor pulmonary embolism affects early outcomes after lung transplantation: a major mechanism of primary graft failure? AB - OBJECTIVE: Primary graft failure remains a significant cause of morbidity and mortality after lung transplantation, and its mechanism is not understood. Previously 2 case reports described fatal primary graft failure due to donor related unexpected pulmonary embolism. This study investigated the incidence, early outcome, and risk factors of unexpected pulmonary embolism in lung transplantation. METHODS: An exploratory retrograde donor lung flush before implantation to diagnose pulmonary embolism (emboli group) or no pulmonary embolism (no-emboli group) was performed in 74 of 122 consecutive lung transplantations. RESULTS: The incidence of macroscopic unexpected pulmonary embolism was 38% (28% clot and 9% fat). In the emboli group, significantly decreased oxygenation (P < .05), increased pulmonary vascular resistance (P < .001), an increased proportion of opacity on chest radiograph (P = .03), prolonged intubation (P < .001) and intensive care unit stay (P < .01), and decreased 1-year survival (P = .03) were seen after transplantation. In multivariate analysis, pulmonary embolism was an independent risk factor for prolonged intubation (hazard ratio, 2.42; P < .01). In logistic regression, death due to trauma with fracture and a smoking history of more than 20 pack-years were significant donor risk factors for pulmonary embolism (adjusted odds ratio, 8.77 and 5.64; P = .02 and .04, respectively). No deleterious effects of the exploratory flush were seen. CONCLUSIONS: Unexpected pulmonary embolism is relatively common, is potentially predicted by donor history (but not by arterial blood gas analysis or chest radiograph), and is associated with primary graft failure. Donor lungs with risk factors of pulmonary embolism should undergo an exploratory flush. When pulmonary embolism is diagnosed, further therapeutic strategies must be considered. PMID- 16256803 TI - Heparin allergy: successful desensitization for cardiopulmonary bypass. PMID- 16256804 TI - Surgical treatment of recurrent transient ischemic attacks and hemoptysis in a young man with multiple pulmonary arteriovenous malformations. PMID- 16256805 TI - Geometric change of mitral valve leaflets and annulus after reconstructive surgery for ischemic mitral regurgitation: real-time 3-dimensional echocardiographic study. PMID- 16256806 TI - Antiphospholipid syndrome and right atrial mass. PMID- 16256807 TI - A Fontan completion through stage I bilateral pulmonary artery banding for hypoplastic left heart syndrome. PMID- 16256808 TI - The association of renal tubular acidosis and cyanotic congenital heart disease. PMID- 16256809 TI - Intraosseous lipoma of the rib. PMID- 16256810 TI - A new management approach for esophageal perforation. PMID- 16256811 TI - Dextrocardia is a component of left-sided Poland syndrome. PMID- 16256812 TI - Posterior reversible encephalopathy syndrome after nontransplant cardiac surgery. PMID- 16256813 TI - Neuronal ultrastructure is preserved by fructose-1,6-bisphosphate after hypothermic circulatory arrest in pigs. PMID- 16256814 TI - Atherosclerosis of radial arterial graft may increase the potential of vessel spasm in coronary bypass surgery. PMID- 16256815 TI - Malignant margin in wedge resection for peripheral lung cancer and adjuvant radiotherapy. PMID- 16256817 TI - Whatever the approach, cutting strut chordae would not smell as sweet. PMID- 16256819 TI - Early anticoagulation after aortic valve replacement with bioprostheses: time to abandon it? PMID- 16256821 TI - The evaluation of aortic atherosclerosis and distensibility in aortic valve stenosis--the role of transesophageal echocardiography. PMID- 16256820 TI - Atrial septal defect in infancy: to close or not to close? PMID- 16256823 TI - Association between cyclooxygenase and epidermal growth factor receptor pathways in non-small cell lung cancer. PMID- 16256826 TI - Information age publishing. PMID- 16256825 TI - Attention shift not memory averaging reduces foveal bias. AB - Two experiments examined which of two mechanisms, attention shift or memory averaging, reduces foveal bias. The target stimulus was a black dot presented for 80 ms while observers maintained fixation. The two main conditions were 'with' and 'without' vertical and horizontal bars as landmarks, which were placed on more eccentric positions than the target stimulus. To induce attention, the landmark was flashed on for 80 ms (Experiment 1) or disappeared (Experiment 2) with a stimulus onset asynchrony of 0, 106.4, or 212.8 ms in both experiments. As a control, non-flashed and non-disappeared landmark conditions were employed. The observers' task was to point to the remembered location of the target with a mouse cursor. The results showed that the magnitudes of foveal bias were significantly lower in the flashed and disappeared landmark conditions than in the without landmark condition. Furthermore, the magnitudes in the flashed and disappeared landmark conditions did not differ from their respective control conditions. The latter finding in the disappeared landmark conditions provides evidence for 'attention shift' against 'memory averaging' as the mechanism reducing foveal bias. PMID- 16256828 TI - Reoperation of a recurrent temporal lobe epilepsy: a technical case report. AB - BACKGROUND: The indications for the reoperation of a recurrent temporal lobe epilepsy, the risks, and outcome have not been well documented. The invasive video electroencephalogram (EEG) monitoring and magnetic resonance imaging (MRI) techniques can reveal the residual tissues and their epileptogenic activity so that a reoperation decision can be made. CASE DESCRIPTION: A 30-year-old patient who had recurrent temporal lobe epilepsy and had undergone 2 operations at the same temporal region is presented. After both of these operations, approximately 6 months later, seizures relapsed. Postoperative neuroimaging studies showed residual mesiotemporal tissues at the operative site. The invasive video EEG monitoring revealed epileptogenic activity originating from these residual tissues. After all of these investigations, it was thought that a third operation was indicated, and the patient was operated. Postoperative course was uneventful. No postoperative deficit was observed. Pathological examination was reported as hippocampal sclerosis. He is seizure-free at his third postoperative year. CONCLUSIONS: Complete resection of epileptogenic mesiotemporal structures at the first operation can prevent the necessity for reoperation in defined cases. The MRI and invasive video EEG monitoring techniques can reveal the residual tissues and their epileptogenic activity in a recurrent epilepsy case. PMID- 16256829 TI - Multiple anterior inferior cerebellar artery aneurysms associated with an arteriovenous malformation: case report. AB - BACKGROUND: Multiple aneurysms of the proximal part of the anterior inferior cerebellar artery (AICA) associated with a distal arteriovenous malformation (AVM) are extremely rare lesions. METHODS: A 52-year-old man was admitted because of sudden headache. Neurological examination revealed ataxia. Computed tomography scan showed a right cerebellar and subarachnoid hemorrhage. Vertebral angiograms demonstrated 3 small aneurysms at the proximal part of the AICA and distal AVM. RESULTS: A right-sided lateral retromastoid suboccipital craniectomy was performed. We observed strangulation and obliteration at the AICA due to multiple clipping for aneurysms. Thus, aneurysms could not be clipped. At 11 years after bleeding, vertebral angiograms showed that 3 aneurysms had slightly enlarged but the AVM remained unchanged. To date, the patient is still doing well except for dizziness. CONCLUSION: This association is very rare at the AICA level, and definitive treatment of this association is sometimes complex. PMID- 16256830 TI - Intracranial multiple tuberculomas: 2 unusual cases. AB - BACKGROUND: Tuberculosis (TB) is still a major public health problem that continues to be an important cause of morbidity and mortality at worldwide level. Only 1% of patients with TB develop an intracranial tuberculoma (Tbm), usually as part of miliary TB. The radiological findings are often nonspecific, and they are difficult to diagnose without an increased index of suspicion. CASES DESCRIPTIONS: In this report, we presented 2 unusual patients with intracranial multiple Tbms, one of whom is a 32-year-old woman who did not have pulmonary TB or other primary origin of TB. Her diagnosis was established histopathologically, and she was treated with antituberculous chemotherapy. The other patient is a 12 year-old girl who has pulmonary TB with portal hypertension together with liver cirrhosis. Her diagnosis was established with clinical, radiological, and laboratory findings, and she was treated with antituberculous chemotherapy for 18 months. CONCLUSIONS: Intracranial Tbms have become rare in developed countries, but TB is still a very common infectious disease at worldwide level. Therefore, diagnosis should be kept in mind when confronted with brain space-occupying lesions, especially in the immunocompromised or malnutritional hosts such as liver cirrhosis. When no other active extracranial tuberculous process is found, the diagnosis should be confirmed by a biopsy before beginning antituberculous treatment, which is rapidly effective. We have not encountered in the literature such a case of multiple intracranial Tbms associated with liver cirrhosis in an adult patient. PMID- 16256832 TI - Acute interhemispheric subdural hematoma due to hemodialysis: case report. AB - BACKGROUND: Acute interhemispheric subdural hematoma (AISH) is an extremely limited complication in patients on maintenance hemodialysis. Diagnostic difficulties result from a fact that symptoms of acute hematoma such as headache, nausea, vomiting, apathy, sleepiness, paresthesia and seizures may also suggest dysequilibrium syndrome, dialytic dementia as well as hypertensive encephalopathy. CASE DESCRIPTION: A case of acute interhemispheric subdural hematoma during long-term dialysis is presented. CONCLUSION: AISH caused by hemodialysis is a very important complication. It is sometimes difficult to differentiate from dysequilibrium or dialysis dementia. Clinicians must pay closer attention on neurological examination of hemodialyzed patient. Surgical treatment must be as prompt as possible because of the possible sudden worsening of the patient's clinical condition. PMID- 16256831 TI - The efficiency of dexamethasone sodium phosphate-encapsulated chitosan microspheres after cold injury. AB - OBJECTIVE: The aims of this study were to evaluate the efficiency of dexamethasone sodium phosphate (DSP) in the treatment of cold injury-induced brain edema and to compare systemic and topical application of DSP. METHODS: Sprague-Dawley rats weighing nearly 300 g were used in the experiments. Brain edema was formed by cold injury using metal sterile rods with a diameter of 4 mm that were previously cooled at -80 degrees C. Twenty-four hours after the injury, animals were decapitated and brain tissues were investigated by wet-dry weight method, lipid peroxidation ratio, and histological examination. RESULTS: The degree of edema was significantly lowered in groups in which DSP was administered using chitosan microspheres and by intraperitoneal route (P < .05). The statistical evaluation of the experimental results was performed using Mann Whitney U test. Histological findings transmission electron microscopy (TEM) correlated with the quantitative results. CONCLUSION: Both intraperitoneal- and microsphere-administered DSP were found to be very effective in a cold injury brain edema model. The authors believe that future studies should lead to new applications of the microsphere formulations prepared by chitosan as the matrix material in many other therapies. PMID- 16256833 TI - History of neurosurgery and neurosurgical applications in Turkey. AB - Although there is evidence of applications of cranial surgery in ancient times, it is commonly accepted that modern surgery started in the late 19th century. The advancements in anesthesiology and aseptic techniques were the main factors contributing to this process. Surgery of the nervous system, however, has a relatively shorter history than surgery of other systems. The process of surgical development in Turkey did not differ from most Western countries. Modern surgery started in 1890 in Turkey. In the beginning, neurosurgical applications were performed by general surgeons. Most of these applications included procedures for craniocerebral traumas and infections and procedures for pain relief. The first neurosurgeon, Dr. Tuner, started working in 1923, operating in some spinal cord and brain tumor and trigeminal neuralgia cases. Other neurosurgeons, Dr. Dilek, Dr. Baydur, and Dr. Kankat, were trained in France and started to work in the mid 1930s. The first neurosurgery department was established in Istanbul in 1923, and the first neurosurgery training program started in the late 1940s. Today, there are more than 50 neurosurgery training centers and more than 500 neurosurgeons in Turkey. There is an increasing number of publications by Turkish neurosurgeons, contributing to the total body of literature in neurosurgery. The current state of neurosurgery in Turkey is parallel to that of the advanced Western countries. PMID- 16256834 TI - Magnesium sulfate treatment decreases caspase-3 activity after experimental spinal cord injury in rats. AB - BACKGROUND: Apoptosis has increasingly been considered as an important factor in secondary injury after spinal cord injury (SCI). Manifestation of apoptotic cell death process involves activation of the caspase-3 apoptotic cascade. The aim of the study was to demonstrate the effect of magnesium sulfate on caspase-3 activity and to compare its effectiveness with methylprednisolone after acute SCI. METHODS: The rats were randomly and blindly allocated into 5 groups of 8 rats each. Spinal cord contusion injury was produced by the weight drop method. The control group consisted of non-injured rats. In the trauma group, no treatment was given, whereas 1 mL saline, 600 mg/kg magnesium sulfate, and 30 mg/kg methylprednisolone sodium succinate (MPSS) were administered in the vehicle and both treatment groups immediately after injury. Twenty-four hours after trauma, spinal cord samples were obtained, and tissue caspase-3 activity levels were examined. A 1-way analysis of variance and the post hoc test were used for statistical analysis. RESULTS: The results showed that caspase-3 activity increased to statistically significantly higher levels in spinal cord after contusion injury than in the control group. Caspase-3 enzyme activity levels were significantly reduced in animals treated either with magnesium sulfate or MPSS. CONCLUSIONS: We have shown that magnesium sulfate decreases caspase-3 activity in rat spinal cord subjected to contusion injury. Magnesium sulfate may have potential therapeutic benefits by reducing apoptotic tissue damage after SCI. PMID- 16256835 TI - MK-801 improves neurological and histological outcomes after spinal cord ischemia induced by transient aortic cross-clipping in rats. AB - BACKGROUND: Glutamergic excitotoxicity has been shown to play a deleterious role in the pathophysiology of ischemic spinal cord injury (ISCI). The aim of this study was to investigate the neuroprotective effect of a single dose of MK-801, an antiexcitotoxic drug, in a rat model of ISCI. METHODS: Ischemic spinal cord injury was induced for 17 minutes in Sprague-Dawley rats using direct aortic arch, just proximal to the left common carotid artery, plus left subclavian artery cross-clamping through a left-sided limited thoracotomy. Study groups were as follows: control group (n = 8) receiving only vehicle and experimental group (n = 8) receiving a single dose of MK-801 (1 mg/kg IV) 10 minutes before aortic clamping. Neurological examination was performed at 6 hours, 24 hours, and daily up to 96 hours. Rats were sacrifice at methylprenisolone socium succinate 96 hours, and spinal cords were removed for histopathology. RESULTS: All the control rats had severe permanent neurological deficits after ISCI, whereas the MK-801 treated rats had statistically (P < .05) better neurological outcome and good recovery. Histopathology revealed severe neuronal necrosis in the lumbar gray matter of control rats, whereas MK-801-treated rats showed mild injury. CONCLUSION: These results demonstrate that combined temporary clipping of the aortic arch (just proximal to the left common carotid artery) plus left subclavian artery for 17 minutes reproduces reliable paraplegia, and a single dose of MK-801 given before ISCI provides significant neuroprotection. PMID- 16256837 TI - Electron microscopic study of the progeny of ependymal stem cells in the normal and injured spinal cord. AB - BACKGROUND: Spinal cord injury (SCI) is a common and often irreversible lesion that can incapacitate patients. Precursor cells in the spinal cord proliferate in response to trauma, and this proliferation can be enhanced by exogenous stimuli such as specific growth factors. In the present study, we examined electron microscopic detection of the proliferation, distribution, and phenotypic fate of these precursor cells in the injured adult rat spinal cord. METHODS: Adult female Sprague-Dawley rats weighing 250 to 300 g were divided into 3 groups. The first group consisted of spinal cord-injured animals with application of a 2.4-g clip extradurally around the spinal cord at the T1 level. A 26-g clip was applied in the second group. The third group included normal uninjured animals. Rats were sacrificed at 3 days, 3 weeks, and 6 weeks after injury. A segment of the spinal cord, 0.4 cm in length, encompassing the injury site was removed and was prepared for electron microscopy. RESULTS: Three days after mild injury (2.4-g clip), ependymal cells had begun to proliferate and had migrated from the central canal. They had a tendency to surround perivascular spaces close to the axons. The central canal rostral to the lesion site was widely dilated at 6 weeks postoperative in the moderately injured groups (26-g clip). The layer of ependymal cells lining the dilated canal showed reduction in cell height. Traumatic syringomyelic cavities were observed in all of the animals. There was an active proliferative response of the ependymal cells to injury. Large clusters of displaced ependymal cells associated with the dilated central canal were observed. Rests of ependymal cells were observed remote from the central canal with a tendency to form rosettes and accessory lumina 6 weeks after trauma. Fascicles of 3 to 8 fibers enclosed within an ependymal cell were a common finding among the ependymal clusters. There were also debris and some ependymal cells in the lumen. CONCLUSION: Trauma induces active proliferation of precursor cells in the ependymal region. These cells may replace neural tissue lost to SCI and may assist in axonal regeneration. PMID- 16256838 TI - The Turkish Neurosurgical Society. PMID- 16256839 TI - Effects of intranigral vs intrastriatal fetal mesencephalic neural grafts on motor behavior disorders in a rat Parkinson model. AB - BACKGROUND: Numerous experimental and clinical studies have shown that intrastriatal fetal mesencephalic grafts grow, survive, and reinnervate host brain tissue, resulting in partial recovery of motor deficits. In addition, pharmacological evidence indicates that these grafts increase dopamine secretion in lesioned brain. However, to date, no grafting method has completely restored the nigrostriatal pathway, and there is no consensus on optimal graft numbers or locations. This study compared outcomes with multiple striatal grafts vs a single intranigral graft in a rat model of Parkinson disease. METHODS: Forty-one female Wistar rats weighing 200 to 250 g were used. First, baseline rotational behavior testing with amphetamine injection was done to identify each animal's dominant nigrostriatal pathway (left vs right hemisphere). Some rats then received a unilateral intranigral injection of 6-hydroxydopamine (4 microL [8 microg]) to produce the Parkinson model lesion, and rotational testing was repeated. One group of the lesioned rats received a single intranigral injection of suspended fetal ventral mesencephalic cells (n = 11), and another received multiple intrastriatal grafts of the same type (n = 11). RESULTS: Both grafted groups showed significant improvement on rotational testing with amphetamine and apomorphine at 6 weeks "postgrafting" (P < .001 for "postlesioning" vs postgrafting results in each of the 2 groups); however, the animals with multiple intrastriatal grafts showed complete recovery from motor asymmetry, whereas the rats with single intranigral grafts showed only partial improvement. CONCLUSION: The findings indicate that multiple intrastriatal grafts result in significantly greater functional improvement than single intranigral grafts in this rat Parkinson model. PMID- 16256840 TI - Diethylstilbestrol-induced prolactinoma: dose-related tumor growth and effect of catecholaminergic cells on prolactin tumor cells. AB - BACKGROUND: Prolactinoma is a pituitary adenoma originating from prolactin secreting epithelial cells of the adenohypophysis. Unfortunately, there appears to be a relatively high recurrence rate despite all pharmacological, radiological, and surgical therapeutic interventions. The aim of the present study was to evaluate the extent of involvement of the dopaminergic dysregulation hypothesis of prolactinomas. We transplanted, in rats, DES-induced prolactinoma cells into the adrenal medulla or under the renal capsule, two tissues rich and poor in catecholaminergic innervation, respectively. METHODS: Prolactinoma was dose-dependently induced in ovariectomized female rats implanted with 10 and 20 mg DES, and tumor cells taken from prolactinoma induced by 20 mg DES were either transplanted under the renal capsule or into the adrenal medulla. RESULTS: Although the adrenal medulla, with its high dopamine content to inhibit prolactin secretion, was devoid of any tumoral development, a significant tumoral development was evident under the renal capsule, seemingly because of no inhibitory control over prolactin secretion coexisting with the dopamine deficiency of the tissue. Results are discussed for an alternatively possible regression and prevention of any relapse of prolactinoma, most possibly occurring because of tuberoinfundibular dopamine deficiency, by the implantation of another dopamine-rich tissue beside the tumoral mass. CONCLUSION: Regression and prevention of any relapse of a tumoral outgrowth, most possibly occurring because of tuberoinfundibular dopamine deficiency, can well be alternatively achieved by the implantation of another dopamine-rich tissue beside the tumoral mass prolactinoma. PMID- 16256841 TI - Arterial vascularization of primary motor cortex (precentral gyrus). AB - BACKGROUND: The precentral gyrus (PG) is the primary motor area and is one of the most eloquent brain regions of neurosurgical interest. Although the arterial supply to the PG is generally known, contributions from different arterial branches such as the anterior cerebral artery (ACA), posterior cerebral artery (PCA), and middle cerebral artery (MCA) have not been comprehensively studied. The aim of the present study was to provide detailed information about the arteries of the PG. METHODS: Twenty adult human brains (40 hemispheres) were obtained, and ACA, MCA, and PCA were separately cannulated and injected with latex. The PG was identified. RESULTS: The ACA supplied the medial one third and the MCA supplied the lateral two thirds of the PG. The PCA did not reach the PG in any of the hemispheres. In 16 hemispheres (40%), the callosomarginal artery and, in 13 hemispheres (32.5%), the pericallosal artery were dominant for the medial one third of the PG. In 11 hemispheres (27.5%), equal dominance was observed. MCA branches at the lateral tip of the PG were classified into precentral, central, and postcentral groups. In 29 hemispheres (72.5%), the central group, and in 4 hemispheres (10%), the precentral group were dominant for the lateral two thirds of the PG. In 7 hemispheres (17.5%), the precentral and central groups were equally dominant. No dominance was identified for the postcentral group. CONCLUSION: In each hemisphere, the PG was supplied by different vascularization patterns of ACA and MCA. The present study is the first to describe and discuss these details. Therefore, awareness of this pattern will provide a great contribution to surgical interventions. PMID- 16256842 TI - Effect of allopurinol in focal cerebral ischemia in rats: an experimental study. AB - BACKGROUND: Allopurinol is a xanthine oxidase inhibitor that prevents the generation of free radicals and may play a role in the protection of the cells during cerebral ischemia. METHODS: We evaluated the protective and therapeutic effect of allopurinol on reversible focal cerebral ischemia-reperfusion model in rats. Cerebral blood flow to the left hemisphere of adult Sprague-Dawley rats (n = 40) was temporarily interrupted by middle cerebral artery (MCA) and bilateral common carotid artery (CCA) occlusion for 3 hours in 5 groups of 8 rats each. Allopurinol (50 mg/kg) was given intraperitoneally 2 hours and immediately before ischemia and immediately and 2 hours after reperfusion in 4 different groups of rats, respectively. Animals were kept alive 24 hours after reperfusion. After sacrifice, infarction volumes and ratios of the brain slices were calculated, and the results were compared with those of the control group. RESULTS: The difference between the allopurinol-administered group and the control group 2 hours before for both infarction volumes and infarction ratios achieved statistical significance. Regarding the allopurinol-administered group immediately before ischemia, infarction volumes and infarction ratios were diminished, but there was no statistically significant difference. The difference between allopurinol-administered and control group immediately after and 2 hours after reperfusion for both infarction volumes and infarction ratios achieved no statistical significance. CONCLUSION: This study showed that allopurinol has a protective effect, but not a therapeutic effect, on cerebral ischemia. PMID- 16256843 TI - In vivo human brain biochemistry after aerobic exercise: preliminary report on functional magnetic resonance spectroscopy. AB - BACKGROUND: Our aim was to disclose whether the positive psychological changes observed after a single bout of aerobic exercise have a biochemical correlate that can be visualized by proton magnetic resonance spectroscopy (MRS) of the human brain. METHODS: Right-handed male volunteers underwent psychological testing and MRS of the frontal lobe of the left hemisphere, both before and after 20 minutes of jogging at about 70% of their maximal aerobic capacity. RESULTS: Although there was a significant decrease on the postexercise anxiety test scores (z = -2.201, P < .05), there was no significant difference between the preexercise and postexercise scores of positive and negative affect. Considering both "amplitude" and "area under the curve" values calculated for the peaks of metabolites N-acetylaspartate (NAA), creatine, and choline, none were found to be significantly changed (P > .05) after the exercise. CONCLUSION: This is, to our knowledge, the first study to report on a functional application of MRS to mood states. Because it offers the ability to directly measure metabolic changes in the brain during neuronal activation, "functional MRS" may be a potential new tool that may be used as an adjunct to functional magnetic resonance imaging. PMID- 16256845 TI - Changing treatment strategy of cavernous sinus meningiomas: experience of a single institution. AB - BACKGROUND: Oncological treatment of a neoplasm is more than surgical removal of the tumor. Probably, this truth is the reason for the ongoing discussion on cavernous sinus meningiomas in the last decade. Debate on optimal management of cavernous sinus meningiomas aims to compare the different treatment strategies: (a) radical surgical resection and (b) conservative surgical resection complemented with radiosurgical treatment. MATERIALS AND METHODS: Natural history of the change in the management strategy of cavernous sinus meningiomas in our department before and after GK facility became available in 1997 allowed us to compare the 2 aforementioned strategies. Before installation of a Leksell GK unit at the hospital in 1997, the neurosurgical team at Marmara University Institute of Neurological Sciences and Faculty of Medicine (Istanbul, Turkey) treated patients with cavernous sinus meningioma using radical resection (radical strategy, group A, 10 patients). After 1997, the same neurosurgical team used understanding of surgical removal of the extracavernous sinus tumor component with GK irradiation of the intracavernous part (conservative strategy, group B, 12 patients). Another group of patients, who were treated with GK as a first-step treatment, was analyzed (GK group, group C, 26 patients). RESULTS: At the end of the third year, more stable tumor volume control was achieved in groups B and C; after the second year, an incline in the tumor volume-time graph was detected. Group B resulted in less cranial nerve-related complications; a certain degree of improvement in cranial nerve deficits was observed. CONCLUSION: Comparing 2 different management strategies for cavernous sinus meningiomas in the same hospital setting using the same neurosurgical group, we conclude that extracavernous resection followed by GK is as effective as radical surgery. Considering cranial nerve complications and third-year tumor volume control achievement, conservative approach yielded better results. Longer follow-up with larger series is necessary. PMID- 16256846 TI - Evaluation of petrosal sinus patency with 3-dimensional contrast-enhanced magnetic resonance venography in petroclival meningiomas for surgical strategy. AB - BACKGROUND: The aim of this study was to perform a detailed anatomical analysis of petroclival venous structures as well as their patencies with 3D contrast enhanced (CE) magnetic resonance venography (MRV) and to identify the potential contribution of these data to the therapeutic approach. METHODS: Ten patients (8 women and 2 men) with unilateral petroclival meningioma were examined using 3D CE MRV in addition to conventional brain protocol. Both coronal source and multiplanar reconstructed images were evaluated for the anatomical orientation. Patency of the cavernous sinus, superior petrosal sinus (SPS), and inferior petrosal sinus (IPS) was assessed. RESULTS: All the patients had a unilateral meningioma (7 on the right and 3 on the left) at the petroclival region. Both SPS and IPS were visualized with adequate intraluminal contrast enhancement in 6 patients, but IPS was absent in 3 on the lesion side, with a patent superior petrosal sinus as the drainage route. One patient had a partially occluded SPS, with IPS being the main course of cavernous sinus drainage. CONCLUSIONS: Cerebral venous anatomy is a challenge to display with noninvasive methods because of flow dynamics, and CE 3D imaging seems to be the modality of choice to evaluate the variational anatomy and patency, which is essential in petroclival meningiomas. Because the cavernous sinus drains into either IPS or SPS, the patent sinus should be protected in surgery if there is tumoral occlusion of the others. PMID- 16256847 TI - Kyphoplasty: 2 years of experience in a neurosurgery department. AB - BACKGROUND: Kyphoplasty is a new technique to treat collapsed vertebral body (VB) fractures. The technique is very effective for achieving rapid pain relief, restoring bone height,and consequent vertebral realignment,and thus stabilization of the vertebra. METHODS: We reviewed 57 patients with vertebral compression fractures. All patients were neurologically intact and presented with severe low back pain or localized pain over the thoracolumbar region. Indications for kyphoplasty were osteoporotic and traumatic compression fractures, osteolytic tumor metastases, and aggressive hemangiomas without spinal canal compression. RESULTS: In 57 patients, 77 levels were treated. Follow-up was 6.5 months. Patients were evaluated with a visual analog scale (VAS) preoperatively and postoperatively. The mean preoperative VAS score was 91.08 for pain, whereas the mean postoperative VAS score was 11.22. Percentage of mean pain relief was 87.9% during the first 6 months of follow-up. We achieved a 43.6% improvement in the height of the compressed VB and 6.3 degrees of improvement in the kyphotic angle. No serious complications occurred. Mean hospital stay was 24 hours. CONCLUSIONS: Kyphoplasty carries significantly less morbidity than vertebroplasty or open surgery. Risk for embolization is low. Normal kyphotic angle can be restored or improved by this technique. In the hands of experienced surgeons, kyphoplasty is a safe and a minimally invasive technique for patients with neoplastic, traumatic, or osteoporotic lesions of the vertebra or sacrum. PMID- 16256848 TI - Hydrocephalus after intraventricular hemorrhage in preterm and low-birth weight infants: analysis of associated risk factors for ventriculoperitoneal shunting. AB - BACKGROUND: Intraventricular hemorrhage and posthemorrhagic hydrocephalus are common causes of neonatal morbidity and mortality among preterm and low-birth weight infants (PT-LBWIs). Clinical management of posthemorrhagic hydrocephalus (PHH) is difficult and not well standardized. In this study, we aimed to determine the incidence of hydrocephalus after intraventricular hemorrhage (IVH) and the associated risk factors for ventriculoperitoneal (V-P) shunting in PT LBWIs. We also aimed to identify the medical-care practices for these babies. METHODS: We reviewed the medical records of 42 babies with IVH diagnosed by cranial ultrasonography (classification of Papile et al, J Pediatr 1978;92:529 34). We compared 11 babies who required a V-P shunt with the 31 control subjects who did not require a V-P shunt or who died before discharge with respect to risk factors involved in V-P shunting. Maternal, perinatal, and neonatal risk factors, and therapies for IVH and PHH were studied as the V-P shunt-associated risk factors. RESULTS: The mean gestational age studied was 28.9 +/- 2.7 weeks, and the mean birth weight was 1164 +/- 391 g. This study revealed an incidence of 26% of PHH in PT-LBWIs with IVH. The most important risk factor for V-P shunt was found to be the severity of IVH (P < .05). Late gestational age and the time of IVH were found to be significant as well (P < .05). The length of hospitalization was found to be longer in patients with V-P shunt (P < .05). Therapies used for IVH and/or PHH were not significant as a risk factor for V-P shunt. In addition, the mortality rate was found to be 38% for all patients with IVH. CONCLUSION: Intraventricular hemorrhage in PT-LBWIs remains a significant problem, particularly when it is associated with PHH leading to long-term neurological impairment and decreased survival rate. PMID- 16256850 TI - An analysis of stereotactic biopsy of brain tumors and nonneoplastic lesions: a prospective clinicopathologic study. AB - BACKGROUND: Appropriate management of progressive, unverified brain lesions should be guided by conclusive pathological diagnosis. Stereotactic biopsy (SB) is established as a less invasive surgical procedure that provides diagnosis. In this prospective study, we analyzed the diagnostic difficulties and risk of SB in the various brain mass lesions, the rate of conclusive pathological diagnosis, and the rate of and the reasons for discrepancy between the intraoperative smear results and conclusive paraffin diagnosis. METHODS: Using computed tomography (CT) and/or magnetic resonance imaging (MRI), 130 cases underwent SB procedure to assess intra-axial brain mass lesions. A CT-MRI fusion and a multiplanar image processing stereotactic program were used in cases who had lesions adjacent to the neurovascular and critical areas. The intraoperative evaluations were made with the smear preparations (SPs) of 1 or 2 biopsy specimens. The conclusive diagnosis was achieved by paraffin preparations of the remainder of the biopsies. The discrepancy between the smear results and the conclusive diagnosis was analyzed. RESULTS: Conclusive histopathologic diagnosis was achieved in 99.23% of the cases. A discrepancy between smear results and conclusive diagnosis was detected in 6.98% of the conclusively diagnosed cases. The major reasons for the discrepancy were necrosis and improper quality of the preparations. There was no mortality, and hemorrhage-related morbidity was observed in 1 case (0.7%). CONCLUSIONS: Necrosis and the improper quality of the smear preparations (SPs) can cause difficulties in establishing a histopathologic diagnosis in SB. Small tissue samples do not decrease the diagnostic yield with the new stereotactic technologies used by an experienced team consisting of a neurosurgeon, pathologist, and radiologist. PMID- 16256851 TI - Destructive stereotactic surgery for treatment of dystonia. AB - BACKGROUND: This study is a retrospective review of the results of stereotactic destructive surgery in selected cases of drug-resistant dystonia. METHODS: Fifty eight patients with drug-resistant dystonia were treated with stereotactic surgery between 1991 and 1999 in our institution. These patients' charts were retrospectively analyzed. The timing of the conducted evaluations was as follows: preoperatively, postoperatively, in the postoperative 1st week, 6th month, 12th month, and also thereafter every year. RESULTS: Symptoms of dystonia occurred before the age of 10 years in 30 patients (51.8%) and after the age of 10 years in 28 patients (48.2%). Generalized dystonia was detected in 41 patients, whereas 11 patients had hemidystonia, 5 patients had focal dystonia, and 1 patient had segmental dystonia. The most common etiologic factor was CP (n = 34). A total of 103 ablative lesions were created in 86 surgical sessions. Thalamotomy, pallidotomy, subthalamotomy, and the region of Forel lesions were performed either separately or in combination. In this series, the mean follow-up time was 102.2 months. Except for 2 cases of temporary hemiparesis, no other complications were observed. Minor improvement was obtained in 17 patients (19.7%), improvement of a medium degree was obtained in 17 patients (19.7%), high-degree improvement was obtained in 11 (12.8%), and very high degree improvement was obtained in 16 (18.6%) patients. A final evaluation revealed permanent improvement in 32 patients (55.2%). CONCLUSION: Production of stereotactic destructive lesions in certain specified targets is a safe method that improves quality of life and aids ambulation in patients with dystonia resistant to medical therapy. PMID- 16256854 TI - ? PMID- 16256853 TI - The trigeminal tract and nucleus procedures in treatment of atypical facial pain. AB - BACKGROUND: Atypical facial pain (AFP) is a throbbing pain situated deep in the eye and malar region, often radiating to the ear, neck, and shoulders. The pain generally is not within any dermatomal or anatomical boundaries. Atypical facial pain is distinct from trigeminal neuralgia and its variants. Therefore, the treatment of AFP should be specified. There is also no consensus in the treatment of AFP. Two different treatment procedures on the trigeminal tract and nucleus in a series of cases with AFP are presented. METHODS: Between 1989 and 2005, 17 patients with AFP, in whom previous therapies had failed, underwent computed tomography (CT)-guided percutaneous trigeminal tractotomy-nucleotomy (TR-NC). One patient with unfavorable response to TR-NC underwent trigeminal dorsal root entry zone (DREZ) operation. RESULTS: In the series with AFP, pain relief was achieved in all of the 17 cases. TR-NC provided maximum to inadequate degrees of pain relief in 16 of 17 patients. Dorsal root entry zone operation provided partial relief in 1 case. Neither mortality nor serious permanent complication was observed in the series. CONCLUSION: Neurosurgical procedures such as TR-NC or trigeminal DREZ operation may be effective in the treatment of intractable AFP. The primary choice of operation should be TR-NC because this procedure is minimally invasive. Trigeminal DREZ operation, which affects a larger spread area, may follow if TR-NC fails. The indications and procedure of choice should be individually tailored, depending on the type of pain, underlying pathology, and experience of the surgeon. PMID- 16256855 TI - ? AB - Evolution of bacterial resistance to antibiotics could be better controlled if we knew how to define the potential of evolution of bacterial resistance towards a given antibiotic, prospectively. No unique method allows to do it. Pieces of answers can be obtained by using diverse methods using bacteriological, ecological pharmaco-epidemiological techniques and experimental models. Association of the results obtained with these various techniques is mandatory to understand the evolution potential of resistance towards a given antibiotic when it is used in the clinical setting. PMID- 16256856 TI - ? AB - Exposure of populations to antibiotics is a condition for the emergence and the diffusion of bacteria resistant to antibiotics. Analysis of epidemiological causality between antibiotic exposure and expansion of antibiotic resistance in human pathogens requires the study of the basis for the emergence and the subsequent epidemic spread of the strains. Emergence of antibiotic resistance under the selective pressure of antibiotics is unavoidable. However, the rapidity of the emergence depends on several parameters including the biochemical mechanism and the genetic support of the resistance and on the couple bacteria/antibiotic considered. Several of these parameters may be studied and measured in vitro. In vivo, relationship between antibiotic exposure and risk for infection or colonisation by pneumococci resistant to beta-lactams or macrolides and methicillin-resistant staphylococci are among the best studied models. In particular for pneumococci, quantitative relationship between antibiotic consumption and resistance, including geographical correlations have been shown but do not allow to establish any causality relationship. Overall, there is no doubt on the question of the impact of antibiotic exposure and the risk of antibiotic resistance. However, the quantification of the risks remains to be accurately studied. PMID- 16256857 TI - ? AB - Resistance is one of failure's reasons. We tried, through clinical experience, to approach the magnitude and nature of the links, between phenotypically defined acquired resistance and clinical failure, in community acquired respiratory infections. An efficient resistance mecanism, able to suppress antibiotic action, is clearely associated to a risk of clinical failure (e.g. betalactamase secretion, target modification using methilation for macrolides, target mutation for fluoroquinoles). Resistance mecanism due to reduction of target affinity (pneumococcus and betalactams) progressively decreasing beta lactam activity depending on its expression, is at present time, not clearely associeted with clinical failure. Critical concentration, defining phenotypical resistance, is predictive of failure if it identifies a bacterial population owning an efficient resistance mecanism. It will not be predictive of failure if that concentration do not detect the resistance mecanism (e.g. parC mutation and levofloxacin) or if the link between antibiotic and resistant bacteria is not binary but depends also on pharmacokinetic parameters (pneumococcus and betalactam). Using resistance as a parametre for antibiotic choice, must integrate several elements: presence or not of a resistance mecanism, type and efficiency of the mecanism, links with clinical failure and antibiotic concentration, type and site of infection. Critical concentration is not allways the magic number that predict failure or success. PMID- 16256858 TI - ? AB - The golden age of antibacterial antibiotics extend from year 1941 to the 1990s decade. At that time, something like an earth quake occurred: from the thirty molecules or so whose development was being achieved or was already marketed, only three were put on the French market, and faced the greatest difficulties to be prescribed by practicians, because: However, while the debate is raging, many of us think "yes we do", as it is a duty to anticipate today the consequences of tomorrow's bacterial resistances. This paper presents three types of propositions to optimise the development of future molecules: The development of new concepts to develop new drugs which would be active against tomorrow's bacteria compels us to manage in a new fashion today's systems, which have reached their own limits. PMID- 16256859 TI - Prospects for personalized cardiovascular medicine: the impact of genomics. AB - Sequencing of the human genome has ushered in prospects for individualizing cardiovascular health care. There is growing evidence that the practice of cardiovascular medicine might soon have a new toolbox to predict and treat disease more effectively. The Human Genome Project has spawned several important "omic" technologies that allow "whole genome" interrogation of sequence variation (re-sequencing, genotyping, comparative genome hybridization), transcription (expression profiling, tissue arrays), proteins (gas or liquid chromatography and tandem mass spectroscopy [MS]), and metabolites (MS or nuclear magnetic resonance profiling); deoxyribonucleic acid, ribonucleic acid, protein, and metabolic approaches all provide more exacting detail of cardiovascular disease mechanisms and, in some cases, are redefining its taxonomy. Pharmacogenomic approaches are emerging across broad classes of cardiovascular therapeutics to assist practitioners in making more precise decisions about which drugs to give to which patients to optimize the benefit-to-risk ratio. Molecular imaging is developing chemical and biological probes that can sense molecular pathway mechanisms that will allow us to monitor health and disease. Together, these tools will enable a paradigm shift from genetic medicine--on the basis of the study of individual inherited characteristics, most often single genes--to genomic medicine, which by its nature is comprehensive and focuses on the functions and interactions of multiple genes and gene products, among themselves and with their environment. The information gained from such analyses, in combination with clinical data, is now allowing us to assess individual risks and guide clinical management and decision-making, all of which form the basis for cardiovascular genomic medicine. PMID- 16256860 TI - Perspectives on dyslipidemia and coronary heart disease in women. AB - Coronary heart disease (CHD) remains the leading cause of death among American women. Numerous differences exist between younger and older women and between women and men with respect to the pathology of CHD and its incidence and prevalence over the life cycle. Differences in lipoprotein levels and lipid fractions play an important role in CHD risk. Hormonal influences on lipoprotein levels in women are complex, change throughout the life span, and are influenced by the administration of oral contraceptives and hormone replacement therapy. Women with obesity, metabolic syndrome, or diabetes have lipid profiles that adversely affect CHD risk. To date, no randomized trials testing the impact of lifestyle changes on lipoprotein levels and subsequent CHD events in non institutionalized women have been performed, and women have not been well represented in clinical end point trials of pharmacologic lipid-lowering therapy. Available evidence suggests that lipid-lowering therapy with statins does provide benefit in reducing the risk of coronary events in women; however, women remain undertreated, and more data are needed to determine optimal cardiovascular prevention and treatment in this population. PMID- 16256861 TI - Safety and efficacy of subcutaneous-only granulocyte-macrophage colony stimulating factor for collateral growth promotion in patients with coronary artery disease. AB - OBJECTIVES: This study was designed to investigate the safety and efficacy of a short-term subcutaneous-only granulocyte-macrophage colony-stimulating factor (GM CSF) protocol for coronary collateral growth promotion. BACKGROUND: The safety and efficacy of an exclusively systemic application of GM-CSF in patients with coronary artery disease (CAD) and collateral artery promotion has not been studied so far. METHODS: In 14 men (age 61 +/- 11 years) with chronic stable CAD, the effect of GM-CSF (molgramostim) on quantitatively assessed collateral flow was tested in a randomized, double-blind, placebo-controlled fashion. The study protocol consisted of an invasive collateral flow index (CFI) measurement in a stenotic as well as a normal coronary artery before and after a two-week period with subcutaneous GM-CSF (10 microg/kg; n = 7) or placebo (n = 7). Collateral flow index was determined by simultaneous measurement of mean aortic, distal coronary occlusive, and central venous pressure. RESULTS: Collateral flow index in all vessels changed from 0.116 +/- 0.05 to 0.159 +/- 0.07 in the GM-CSF group (p = 0.028) and from 0.166 +/- 0.06 to 0.166 +/- 0.04 in the placebo group (p = NS). The treatment-induced difference in CFI was +0.042 +/- 0.05 in the GM-CSF group and -0.001 +/- 0.04 in the placebo group (p = 0.035). Among 11 determined cytokines, chemokines, and their monocytic receptor concentrations, the treatment induced change in CFI was predicted by the respective change in tumor necrosis factor-alpha concentration. Two of seven patients in the GM-CSF group and none in the placebo group suffered an acute coronary syndrome during the treatment period. CONCLUSIONS: A subcutaneous-only, short-term protocol of GM-CSF is effective in promoting coronary collateral artery growth among patients with CAD. However, the drug's safety regarding the occurrence of acute coronary syndrome is questionable. PMID- 16256862 TI - Outcomes and risks of granulocyte colony-stimulating factor in patients with coronary artery disease. AB - OBJECTIVES: Cytokine mobilization of progenitor cells from bone marrow may promote myocardial neovascularization with relief of ischemia. BACKGROUND: Patients with coronary artery disease (CAD) have low numbers of endothelial progenitor cells compared with healthy subjects. METHODS: Granulocyte colony stimulating factor (G-CSF), 10 microg/kg/day for five days, was administered to 16 CAD patients. Progenitor cells were measured by flow cytometry; ischemia was assessed by exercise stress testing and by dobutamine stress cardiac magnetic resonance imaging. RESULTS: Granulocyte colony-stimulating factor increased CD34+/CD133+ cells in the circulation from 1.5 +/- 0.2 microl to 52.4 +/- 10.4 microl (p < 0.001), similar to the response observed in 15 healthy subjects (75.1 +/- 12.6 microl, p = 0.173). Indices of platelet and coagulation activation were not changed by treatment, but C-reactive protein increased from 4.5 +/- 1.3 mg/l to 8.6 +/- 1.3 mg/l (p = 0.017). Two patients experienced serious adverse events: 1) non-ST-segment elevation myocardial infarction (MI) 8 h after the fifth G-CSF dose, and 2) MI and death 17 days after treatment. At 1 month after treatment, there was no improvement from baseline values (i.e., reduction) in wall motion score (from 25.7 +/- 2.1 to 28.3 +/- 1.9, p = 0.196) or segments with abnormal perfusion (7.6 +/- 1.1 to 7.7 +/- 1.1, p = 0.916) and a trend towards a greater number of ischemic segments (from 4.5 +/- 0.6 to 6.1 +/- 1.0, p = 0.068). There was no improvement in exercise duration at 1 month (p = 0.37) or at 3 months (p = 0.98) versus baseline. CONCLUSIONS: Granulocyte colony-stimulating factor administration to CAD patients mobilizes cells with endothelial progenitor potential from bone marrow, but without objective evidence of cardiac benefit and with the potential for adverse outcomes in some patients. PMID- 16256863 TI - Granulocyte colony-stimulating factor and granulocyte-macrophage colony stimulating factor: double-edged swords. PMID- 16256864 TI - Regeneration of human infarcted heart muscle by intracoronary autologous bone marrow cell transplantation in chronic coronary artery disease: the IACT Study. AB - OBJECTIVES: Stem cell therapy may be useful in chronic myocardial infarction (MI); this is conceivable, but not yet demonstrated in humans. BACKGROUND: After acute MI, bone marrow-derived cells improve cardiac function. METHODS: We treated 18 consecutive patients with chronic MI (5 months to 8.5 years old) by the intracoronary transplantation of autologous bone marrow mononuclear cells and compared them with a representative control group without cell therapy. RESULTS: After three months, in the transplantation group, infarct size was reduced by 30% and global left ventricular ejection fraction (+15%) and infarction wall movement velocity (+57%) increased significantly, whereas in the control group no significant changes were observed in infarct size, left ventricular ejection fraction, or wall movement velocity of infarcted area. Percutaneous transluminal coronary angioplasty alone had no effect on left ventricular function. After bone marrow cell transplantation, there was an improvement of maximum oxygen uptake (VO2max, +11%) and of regional 18F-fluor-desoxy-glucose uptake into infarct tissue (+15%). CONCLUSIONS: These results demonstrate that functional and metabolic regeneration of infarcted and chronically avital tissue can be realized in humans by bone marrow mononuclear cell transplantation. PMID- 16256865 TI - Bone marrow cell-mediated cardiac regeneration a veritable revolution. PMID- 16256866 TI - Granulocyte colony-stimulating factor and stem cell factor improve contractile reserve of the infarcted left ventricle independent of restoring muscle mass. AB - OBJECTIVES: We investigated whether granulocyte colony-stimulating factor (G-CSF) and stem cell factor (SCF) could promote myocardial regeneration after coronary artery occlusion and improve left ventricular (LV) function. BACKGROUND: Cytokine induced mobilization of bone marrow stem cells in the heart may represent a promising strategy for replacing infarcted myocardium. METHODS: Sprague-Dawley rats were subjected to permanent coronary occlusion. A treated group (n = 19) received G-CSF (100 microg/kg) and SCF (25 microg/kg) subcutaneously, starting 2 h after surgery and continuing daily for an additional 4 days. Control rats (n = 21) received sterile water. The peripheral blood content in hematopoietic progenitor cells was analyzed. RESULTS: At eight weeks, LV angiograms (rest and dobutamine stress) and histologic analysis were performed. At rest, LV ejection fraction (LVEF) was 0.45 in controls and 0.52 in treated hearts (p = 0.16). For any infarct size, LVEF was greater in the treated group (p = 0.045). Under dobutamine stress, treated animals had smaller LV end-diastolic and -systolic volumes (0.37 +/- 0.04 ml and 0.16 +/- 0.03 ml) versus control animals (0.51 +/- 0.05 ml and 0.26 +/- 0.04 ml; p = 0.026 and 0.048) with a 7% improvement in ejection fraction. Scar thickness was 1.1 +/- 0.1 mm in treated hearts and 1.0 +/ 0.1 mm in controls (p = 0.36). Scar morphology was similar in both groups without obvious new muscle in the scar. CONCLUSIONS: Because we did not find evidence of new muscle cells in the infarct area, our conclusion is that G-CSF and SCF enhanced the LV functional reserve of the heart without replacing scar tissue. PMID- 16256867 TI - Evaluation of four-year coronary artery response after sirolimus-eluting stent implantation using serial quantitative intravascular ultrasound and computer assisted grayscale value analysis for plaque composition in event-free patients. AB - OBJECTIVES: This study sought to evaluate the long-term arterial response after sirolimus-eluting stent implantation. BACKGROUND: Sirolimus-eluting stents are effective in inhibiting neointimal hyperplasia without affecting plaque volume behind the stent struts at six months. METHODS: Serial quantitative intravascular ultrasound and computer-assisted grayscale value analysis over four years were performed in 23 event-free patients treated with sirolimus-eluting stents. RESULTS: In the first two years, the mean plaque volume (155.5 +/- 42.8 mm3 post procedure and 156.8 +/- 57.7 mm3 at two years, p = 0.86) and plaque compositional change expressed as mean percent hypoechogenic tissue of the plaque behind the stent struts (78.9 +/- 8.6% post-procedure and 78.2 +/- 8.9% at two years, p = 0.67) did not significantly change. However, significant plaque shrinking (change in plaque volume = -18.4 mm3, p = 0.02) with an increase in plaque echogenicity (change in percent hypoechogenic tissue = -7.8%, p < 0.0001) was observed between two and four years. The mean neointimal volume increased over four years from 0 to 8.4 +/- 5.8 mm3 (p < 0.0001). However, no further statistically significant change occurred between two and four years (7.0 +/- 6.7 mm3 vs. 8.4 +/- 5.8 mm3, p = 0.25). CONCLUSIONS: Between two and four years after sirolimus-eluting stent implantation, peri-stent tissue shrank with a concomitant increase in echogenicity. These intravascular ultrasound findings suggest that late chronic artery responses may evolve for up to four years after sirolimus-eluting stent implantation. In addition, the fact that the neointima does not significantly change from two to four years may suggest that the biological phenomenon of a delayed healing response has begun to subside. PMID- 16256868 TI - Randomized evaluation of the TriActiv balloon-protection flush and extraction system for the treatment of saphenous vein graft disease. AB - OBJECTIVES: The Protection During Saphenous Vein Graft Intervention to Prevent Distal Embolization (PRIDE) study compared outcomes with the TriActiv System (Kensey Nash Corp., Exton, Pennsylvania), a balloon-protection flush and extraction device, with an embolic protection group during treatment of saphenous venous grafts (SVGs). BACKGROUND: Treatment of SVGs with embolic protection reduces adverse cardiac events. METHODS: We conducted a prospective trial randomizing 631 patients with coronary ischemia and lesions in SVGs to embolic protection with the TriActiv System or control group (Guardwire System [Medtronic AVE, Santa Rosa, California] or Filterwire EX [Boston Scientific Corp., Maple Grove, Minnesota]). RESULTS: The incidence of major adverse cardiac events at 30 days was 11.2% for the TriActiv group and 10.1% for the control group (relative risk = 1.1%; 95% confidence interval 0.67 to 1.76; p = 0.65; p = 0.02 for non inferiority). Safety and efficacy end points were similar between groups except that patients randomized to the TriActiv System had more hemorrhagic complications (10.9% vs. 5.4%; p = 0.01). CONCLUSIONS: The TriActiv System was not inferior to approved embolic protection devices for the treatment of diseased SVGs. PMID- 16256869 TI - A new anatomic score for prognosis after cardiac catheterization in patients with previous bypass surgery. AB - OBJECTIVES: The purpose of this study was to determine the value of a new anatomic score for prognosis after diagnostic catheterization in patients with previous coronary artery bypass grafting (CABG). BACKGROUND: Previous CABG patients comprise a growing proportion of patients with coronary artery disease (CAD). Whereas prognostic scores are available to adjust for native CAD, there are no comparable scores for patients with previous CABG. METHODS: We studied 3,178 previous CABG patients (2,729 in a training set) who underwent cardiac catheterization. With a Cox model to develop relative weights in the training set, we created a graft index that adjusted native anatomy for territories with grafts free of significant (> or =75%) stenoses. Scaling the regression coefficients by the maximum coefficient created an index ranging from 0 to 100, where 100 was three-vessel CAD with no patent grafts. RESULTS: The graft index was significantly associated with all-cause death (chi-square = 121.9, p < 0.001). In combined models, the index was more strongly associated with all-cause death than either number of diseased vessels (chi-square = 68.0 and 1.7, respectively) or the Duke CAD index (chi-square = 54.3 and 9.5, respectively). In models for death using an independent validation set, the index was also associated more strongly than either native disease descriptors. In a model including other clinical variables, the graft index remained significantly associated with all-cause death (chi-square = 40.1, p < 0.001). CONCLUSIONS: For previous CABG patients, the Duke graft index was significantly more associated with prognosis than native anatomy alone and quantifies the effect of patent grafts on survival. This tool has the potential to help determine prognosis and inform the referral of post-CABG patients to repeat revascularization procedures. PMID- 16256870 TI - Suppression of endothelial progenitor cells in human coronary artery disease by the endogenous nitric oxide synthase inhibitor asymmetric dimethylarginine. AB - OBJECTIVES: We tested the hypothesis that asymmetric dimethylarginine (ADMA) may be an endogenous inhibitor of endothelial progenitor cells (EPCs). BACKGROUND: Endothelial progenitor cells play a pivotal role in regeneration of injured endothelium, thereby limiting the formation of atherosclerotic lesions. Reduced numbers of EPCs may affect progression of coronary artery disease. Regulation of EPC mobilization and function is mediated in part by nitric oxide (NO). Endogenous inhibitors of NO synthases, such as ADMA, contribute to endothelial dysfunction and injury. METHODS: We used flow cytometry and in vitro assays to investigate the relationship between EPC number and function with ADMA plasma levels in patients with stable angina. RESULTS: The plasma concentration of ADMA was related to the severity of coronary artery disease and correlated inversely with the number of circulating CD34+/CD133+ progenitor cells (r = -0.69; p < 0.0001) and endothelial colony forming units (CFUs) (r = -0.75; p < 0.0001). Adjusting for all patient characteristics, we confirmed these findings in multivariate regression analyses. In vitro differentiation of EPCs was repressed by ADMA in a concentration-dependent manner. Compared with untreated cells, ADMA reduced EPC incorporation into endothelial tube-like structures to 27 +/- 11% (p < 0.001). Asymmetric dimethylarginine repressed the formation of CFUs from cultured peripheral blood mononuclear cells to 35 +/- 7% (p < 0.001). Asymmetric dimethylarginine decreased endothelial nitric oxide synthase activity in EPCs to 64 +/- 6% (p < 0.05) when compared with controls. Co-incubation with the hydroxymethyl glutaryl coenzyme A reductase inhibitor rosuvastatin abolished the detrimental effects of ADMA. CONCLUSIONS: Asymmetric dimethylarginine is an endogenous inhibitor of mobilization, differentiation, and function of EPCs. This contributes to the cardiovascular risk in patients with high ADMA levels and may explain low numbers and function of EPCs in patients with coronary artery disease. PMID- 16256871 TI - ADMAring Endothelial Progenitor Cells Accident, Association, or Antecedent. PMID- 16256872 TI - Overestimation of platelet aspirin resistance detection by thrombelastograph platelet mapping and validation by conventional aggregometry using arachidonic acid stimulation. AB - OBJECTIVES: This study sought to determine the prevalence of platelet aspirin resistance using methods that directly indicate the degree of platelet cyclooxygenase inhibition. BACKGROUND: Aspirin resistance in platelets may be overestimated by nonspecific laboratory measurements that do not isolate cyclooxygenase activity. METHODS: Arachidonic acid (AA)-induced light transmittance platelet aggregation (LTA) and thrombelastography (TEG) platelet mapping were performed on the blood of healthy subjects (n = 6) before and 24 h after receiving 325 mg aspirin, and on 223 patients reporting compliance with long-term daily aspirin treatment (n = 203 undergoing percutaneous intervention [PCI] and n = 20 with a history of stent thrombosis). Aspirin resistance was defined as >20% aggregation by LTA or >50% aggregation by TEG. RESULTS: In healthy subjects, AA-induced aggregation by LTA was 82 +/- 10% before and 2 +/- 1% at 24 h after aspirin (p < 0.001), and aggregation by TEG was 86 +/- 14% before and 5 +/- 7% at 24 h after aspirin (p < 0.001). In compliant patients, AA induced aggregation by LTA was 3 +/- 2% before PCI and 3 +/- 2% after PCI (p = NS), and aggregation by TEG was 5 +/- 9% before PCI and 6 +/- 14% after PCI (p = NS). Seven PCI patients were noncompliant, and all were aspirin sensitive after in-hospital aspirin treatment. Among 223 patients, only one patient ( approximately 0.4%) was resistant to aspirin treatment. CONCLUSIONS: Platelet aspirin resistance assessed by methods that directly indicate inhibition of cyclooxygenase is rare in compliant patients with coronary artery disease. PMID- 16256873 TI - On defining aspirin resistance. PMID- 16256875 TI - Association of myocardial ischemia with mortality and implantable cardioverter defibrillator therapy in patients with coronary artery disease at risk of arrhythmic death. AB - OBJECTIVES: We sought to assess the relation between myocardial ischemia during stress echocardiography and major events in patients with implantable cardioverter-defibrillator (ICD). BACKGROUND: The association of myocardial ischemia with subsequent ICD therapy and mortality is unknown. METHODS: We studied 90 patients (age 65 +/- 13 years, 27 women) with history of coronary heart disease who received ICD for primary (53 patients) or secondary (37 patients) prevention of sudden cardiac death. Sixty-five (72%) patients had a previous coronary artery bypass surgery. Patients underwent exercise treadmill or dobutamine stress echocardiography. Ischemia was defined as new or worsening wall motion abnormalities. End points were death and appropriate ICD therapy. RESULTS: Mean ejection fraction was 34 +/- 12%. During a mean follow-up of 2.8 +/- 1.5 years, 5 patients died and 19 patients had ICD therapy. Ischemia was detected in 20 of 24 patients with subsequent events and in 24 of 66 patients without (83% vs. 36%, p < 0.001). Events occurred in 17 of the 32 patients (53%) with both ischemia and inducible ventricular tachycardia (VT) on electrophysiologic (EP) studies. None of the 16 patients without ischemia or inducible VT on EP studies had events. In a Cox multivariate analysis model, independent predictors of events were a history of spontaneous sustained VT (hazard ratio [HR] 1.9, 95% confidence interval [CI] 1.3 to 3.8), inducible VT on EP studies (HR 1.7, 95% CI 1.2 to 4.5), and ischemia (HR 2.1, 95% CI 1.2 to 3.5). CONCLUSIONS: Ischemia during stress echocardiography is an independent predictor of death and ICD therapy in patients with coronary heart disease at high risk of arrhythmic death. Patients without inducible ischemia or VT on a previous EP study have a very low risk of events. A combination of ischemia and a positive EP study is associated with a very high risk of events. PMID- 16256874 TI - Factors influencing appropriate firing of the implanted defibrillator for ventricular tachycardia/fibrillation: findings from the Multicenter Automatic Defibrillator Implantation Trial II (MADIT-II). AB - OBJECTIVES: The purpose of this study was to prospectively examine the role of clinical, laboratory, echocardiographic, and electrophysiological variables as predictors of appropriate initial implantable cardioverter-defibrillator (ICD) therapy for ventricular tachycardia (VT) or ventricular fibrillation (VF) or death in the Multicenter Automatic Defibrillator Implantation Trial II (MADIT-II) population. BACKGROUND: There is limited information regarding the determinants of appropriate ICD therapy in patients with reduced ventricular function after a myocardial infarction. METHODS: We used secondary analysis in one arm of a multicenter randomized clinical trial in patients with a previous myocardial infarction and reduced left ventricular function. RESULTS: We analyzed baseline and follow-up data on 719 patients enrolled in the ICD arm of the MADIT-II study. Appropriate ICD therapy was observed in 169 subjects. Clinical, laboratory, echocardiographic, and electrophysiological variables, along with measures of clinical instability such as interim hospitalization for congestive heart failure (IH-CHF) and interim hospitalization for coronary events (IH-CE), were examined with proportional hazards models and Kaplan-Meier time-to-event curves before and after first interim hospitalization. Interim hospitalization-CHF, IH-CE, no beta blockers, digitalis use, blood urea nitrogen (BUN) >25, body mass index (BMI) > or =30 kg/m2, and New York Heart Association functional class >II were associated with increased risk for appropriate ICD therapy for VT, VF, or death. In a multivariate (stepwise selection) analysis, IH-CHF was associated with an increased risk for the end point of either VT or VF (hazard ratio [HR] 2.52, 95% confidence interval [CI] 1.69 to 3.74, p < 0.001) and for the combined end point of VT, VF, or death (HR 2.97, 95% CI 2.15 to 4.09, p < 0.001). Interim hospitalization-CE was associated with an increased risk for VT, VF, or death (HR 1.66, 95% CI 1.09 to 2.52, p = 0.02). CONCLUSIONS: These results provide important mechanistic information, suggesting that worsening clinical condition and cardiac instability, as reflected by an IH-CHF or IH-CE, are subsequently associated with a significant increase in the risk for appropriate ICD therapy (for VT/VF) and death. PMID- 16256876 TI - Who needs a defibrillator? The beat goes on. PMID- 16256877 TI - Hospitalized patients with atrial fibrillation and a high risk of stroke are not being provided with adequate anticoagulation. AB - OBJECTIVES: The purpose of this study was to determine both treatment gaps and predictors of warfarin use in atrial fibrillation (AF) patients enrolled in a national multicenter study. BACKGROUND: The National Anticoagulation Benchmark Outcomes Report (NABOR) is a performance improvement program designed to benchmark anticoagulation prophylaxis, treatment, and outcomes among participating hospitals. METHODS: A retrospective cohort study of inpatients was performed at 21 teaching, 13 community, and 4 Veterans Administration hospitals in the U.S. Patients with an ICD-9-CM code for AF (427.31) were randomly selected. RESULTS: Among the 945 patients studied, the mean age was 71.5 (+/- 13.5) years; 43% were >75 years of age, 54.5% were men, and 67% had a history of hypertension. Most (86%) had factors that stratified them as at high risk of stroke, and only 55% of those received warfarin. Neither warfarin nor aspirin were prescribed in 21% of high-risk patients, including 18% of those with a previous stroke, transient ischemic attack, or systemic embolic event. Age >80 years (p = 0.008) and perceived bleeding risk (p = 0.022) were negative predictors of warfarin use. Persistent/permanent AF (p < 0.001) and history of stroke, transient ischemic attack, or systemic embolus (p = 0.014) were positive predictors of warfarin use, whereas high-risk stratification was not. CONCLUSIONS: This study confirms the under-use of warfarin, but also adds to published reports in several regards. It showed that risk stratification, the guidepost for treatment in international guidelines, had little effect on warfarin use, and that age >80 years and AF classification (permanent/persistent) are factors that influence warfarin use. PMID- 16256878 TI - Lifelong left ventricular remodeling of hypertrophic cardiomyopathy caused by a founder frameshift deletion mutation in the cardiac Myosin-binding protein C gene among Japanese. AB - OBJECTIVES: We studied the longitudinal evolution of hypertrophic cardiomyopathy (HCM) caused by a founder frameshift mutation in the cardiac myosin-binding protein C (MyBPC) gene. BACKGROUND: Mutations in the MyBPC gene have been associated with delayed expression of HCM and a good prognosis. Few studies, however, demonstrated the phenotype-genotype correlations in the longitudinal study. METHODS: We studied long-term evolution of clinical features of 15 unrelated families who were found to have an identical frameshift mutation in the MyBPC gene: a one-base deletion of a thymidine at nucleotide 11645 (V592fs/8). RESULTS: Thirty-nine individuals in 15 families were genotype-positive. Thirty of the 39 individuals with the mutation were phenotype-positive. The disease penetrance was 100% in subjects > or =50 years and 65% in those <50 years. "End stage" HCM (ejection fraction <50%) was observed in 7 (18%) of the 39 genotype positive individuals (7 [23%] of the 30 phenotype-positive patients); 6 of them were 60 years or older. Seven patients were hospitalized for treatment of repeated congestive heart failure, and four patients died or had implantable cardioverter-defibrillator discharge (13%; incidence, 1.4%/year) during a mean follow-up period of 9.2 +/- 5.5 years. CONCLUSIONS: Elderly patients with a V592fs/8 mutation in the MyBPC gene may evolve into the "end-stage" HCM, characterized by left ventricular systolic dysfunction, cavity dilation, and irreversible heart failure. The clinical course in patients with this mutation is not benign in the long run, with progressive left ventricular remodeling with advancing age. PMID- 16256879 TI - Are longitudinal, natural history studies the next step in genotype-phenotype translational genomics in hypertrophic cardiomyopathy? PMID- 16256880 TI - Usefulness of a comprehensive cardiovascular magnetic resonance imaging assessment for predicting recovery of left ventricular wall motion in the setting of myocardial stunning. AB - OBJECTIVES: We sought to evaluate the usefulness of a comprehensive assessment of four cardiovascular magnetic resonance imaging (CMR)-derived myocardial viability indexes in the setting of myocardial stunning. BACKGROUND: Cardiovascular magnetic resonance imaging allows the simultaneous assessment of several viability indexes. METHODS: We studied 40 patients with a first ST-segment elevation myocardial infarction (MI) and an open infarct-related artery. At the first week, using CMR, wall motion (WM), and four viability indexes were determined: wall thickness, WM improvement with low-dose dobutamine, perfusion, and transmural extent of necrosis. We created a comprehensive score based on the presence and the relative power of these viability indexes for predicting normal WM at the sixth month. RESULTS: Of 153 dysfunctional segments at the first week, 59 (39%) exhibited normal WM at the sixth month. According to the odds ratio of viability indexes for predicting normal WM, we developed a five-level predictive score. The proportions of segments showing normal WM at sixth month were as follows; Level 1 (0 indexes): 0 of 13 (0%); Level 2 (normal thickness and/or perfusion): 14 of 82 (17%); Level 3 (dobutamine response): 5 of 11 (45%); Level 4 (non-transmural necrosis): 20 of 26 (77%); Level 5 (non-transmural necrosis and dobutamine response): 20 of 21 (95%), p < 0.0001 for the trend. These proportions were similar in a matched prospective validation group comprising 16 patients (0%, 18%, 62%, 77%, and 90% for levels 1 to 5, respectively, p < 0.0001 for the trend). CONCLUSIONS: A comprehensive analysis of the four more widely used CMR derived viability indexes is useful for predicting late systolic function after myocardial infarction. PMID- 16256881 TI - Normal vascular aging: differential effects on wave reflection and aortic pulse wave velocity: the Anglo-Cardiff Collaborative Trial (ACCT). AB - OBJECTIVES: The aim of the current investigation was to test the hypothesis that age-related changes in augmentation index (AIx) are more prominent in younger individuals (<50 years), whereas changes in aortic stiffness per se are more marked in older individuals (>50 years). BACKGROUND: Aging exerts a number of deleterious changes in the cardiovascular system, and, in particular, on the large arteries. Previous studies have suggested that AIx and pulse wave velocity (PWV) increase linearly with age, yet epidemiological data concerning pulse pressure suggest that large artery stiffening predominantly occurs later in life. METHODS: Peripheral and central blood pressure, augmentation pressure (AP), and AIx were determined in 4,001 healthy, normotensive individuals, aged 18 to 90 years. Aortic and brachial PWV were also determined in a subset of 998 subjects. RESULTS: Peripheral and central pulse pressure, AP, AIx, and aortic and brachial PWV all increased significantly with age; however, the age-related changes in AIx and aortic PWV were non-linear, with AIx increasing more in younger individuals, whereas the changes in PWV were more prominent in older individuals. CONCLUSIONS: These data suggest that AIx might be a more sensitive marker of arterial stiffening and risk in younger individuals but aortic PWV is likely to be a better measure in older individuals. PMID- 16256883 TI - Patent foramen ovale: current pathology, pathophysiology, and clinical status. AB - Patent foramen ovale (PFO) is experiencing increased clinical interest as a congenital cardiac lesion persisting into adulthood. It is implicated in several serious clinical syndromes, including stroke, myocardial infarction, and systemic embolism. The PFO is now amenable to percutaneous interventional therapies, and multiple novel technologies are either available or under development for lesion closure. The PFO should be better understood to take advantage of emerging percutaneous treatment options. This paper reviews PFO anatomy, pathology, pathophysiology, and clinical impact and discusses current therapeutic options. PMID- 16256884 TI - President's page: Prevention's place in cardiology: what the future holds. PMID- 16256882 TI - Walking and sports participation and mortality from coronary heart disease and stroke. AB - OBJECTIVES: We aimed to examine the impact of exercise on mortality from cardiovascular disease (CVD) in Asian populations. BACKGROUND: Few data have been available in Asian countries, where job-related physical activity is higher than that in Western countries. METHODS: Between 1988 and 1990, 31,023 men and 42,242 women in Japan, ages 40 to 79 years with no history of stroke, coronary heart disease (CHD), or cancer, completed a self-administered questionnaire. Systematic mortality surveillance was performed through 1999, and 1,946 cardiovascular deaths were identified. We chose the second lowest categories of walking and sports participation as the reference to reduce a potential effect of ill health. RESULTS: Men and women who reported having physical activity in the highest category (i.e., walking > or =1 h/day or doing sports > or =5 h/week) had a 20% to 60% lower age-adjusted risk of mortality from CVD, compared with those in the second lowest physical activity category (i.e., walking 0.5 h/day, or sports participation for 1 to 2 h/week). Adjustment for known risk factors, exclusion of individuals who died within two years of baseline inquiry, or gender-specific analysis did not substantially alter these associations. The multivariate adjusted hazard ratios (95% confidence interval) for the highest versus the second lowest categories of walking or sports participation were 0.71 (0.54 to 0.94) and 0.80 (0.48 to 1.31), respectively, for ischemic stroke (IS); 0.84 (0.64 to 1.09) and 0.51 (0.32 to 0.82), respectively, for CHD; and 0.84 (0.75 to 0.95) and 0.73 (0.60 to 0.90), respectively, for CVD. CONCLUSIONS: Physical activity through walking and sports participation might reduce the risk of mortality from IS and CHD. PMID- 16256885 TI - Noninvasive measurement of cardiac output during exercise by inert gas rebreathing technique: a new tool for heart failure evaluation. PMID- 16256886 TI - Can perindopril delay the onset of heart failure in duchenne muscular dystrophy? PMID- 16256888 TI - Concomitant surgical myocardial revascularization is still the gold standard for combined valvular and coronary heart disease. PMID- 16256890 TI - Drug crime is a source of abused pain medications in the United States. PMID- 16256891 TI - Intravenous infusion of midazolam and flunitrazepam for insomnia on Japanese palliative care units. PMID- 16256893 TI - Topical morphine in the treatment of painful ulcers. PMID- 16256892 TI - Incidence of chronic pain in patients with end-stage renal disease on dialysis. PMID- 16256894 TI - Musical hallucinations and opioids: a word of caution. PMID- 16256895 TI - Ethical validity of palliative sedation therapy: a multicenter, prospective, observational study conducted on specialized palliative care units in Japan. AB - Although palliative sedation therapy is often required in terminally ill cancer patients to achieve acceptable symptom relief, empirical data supporting the ethical validity of this approach are lacking. The primary aim of this study was to systematically investigate whether empirical evidence supports the ethical validity of sedation. This was a multicenter, prospective, observational study, which was conducted by 21 specialized palliative care units in Japan. One-hundred two consecutive adult cancer patients who received continuous deep sedation were enrolled. Continuous deep sedation was defined as the continuous use of sedative medications to relieve intolerable and refractory distress by achieving almost or complete unconsciousness until death. Prior to the study, we conceptualized the ethical validity of sedation from the viewpoints of physicians' intent, proportionality, and autonomy. Sedation was performed mainly with midazolam and phenobarbital. The initial doses of midazolam and phenobarbital were 1.5 mg/hour and 20 mg/hour, respectively. Main administration routes were continuous subcutaneous infusion and continuous intravenous infusion, and no rapid intravenous injection was reported. Of 59 patients who received artificial hydration or could intake adequate fluids/foods orally before sedation, 63% received artificial hydration therapy after sedation, and in the remaining patients, artificial hydration was withheld or withdrawn due to fluid retention symptoms and/or patient wishes. Of 66 patients who were able to verbally express themselves, 95% explicitly stated that symptoms were intolerable. The etiologies of the symptoms requiring sedation were primarily related to the progression of the underlying malignancy, such as cancer cachexia and organ failure, and standard palliative treatments had failed: steroids in 68% of patients with fatigue, opioids in 95% of patients with dyspnea, antisecretion medications in 75% of patients with bronchial secretion, antipsychotic medications in 74% of patients with delirium, and opioids in all patients with pain. On the basis of the Palliative Prognostic Index, 94% of the patients were predicted to die within 3 weeks. Before sedation, 67% of the patients expressed explicit wishes for sedation. In the remaining 34 patients, previous wishes for sedation were noted in 4 patients, and in the other 30 patients, the families were involved in the decision-making process. The chief reason for patient non-involvement in the decision making was cognitive impairment. These data indicate that palliative sedation therapy performed in specialized palliative care units in Japan generally followed the principles of double effect, proportionality, and autonomy. PMID- 16256896 TI - Efficacy and safety of palliative sedation therapy: a multicenter, prospective, observational study conducted on specialized palliative care units in Japan. AB - Although palliative sedation therapy is often required in terminally ill cancer patients, its efficacy and safety are not sufficiently understood. The primary aims of this multicenter observational study were to 1) explore the efficacy and safety of palliative sedation therapy, and 2) identify the factors contributing to inadequate symptom relief and complications, using a prospective study design, clearly defined measurement methods, and a consecutive sample from 21 specialized palliative care units in Japan. A sample of 102 consecutive adult cancer patients who received continuous deep sedation were enrolled. Physicians prospectively evaluated the intensity of patient symptoms, communication capacity, respiratory rate, and complications related to sedation. Symptoms were measured on the Agitation Distress Scale, the Memorial Delirium Assessment Scale, and the ad hoc symptom severity scale (0 = no symptoms, 1 = mild and tolerable symptoms, 2 = intolerable symptoms for less than 15 minutes in the previous one hour, and 3 = intolerable symptoms continuing for more than 15 minutes in the previous one hour). Inadequate symptom relief was defined as presence of hyperactive delirium (item 9 of the Memorial Delirium Assessment Scale >or=2) or grade 2 or 3 symptom intensity 4 hours after sedation. The degree of communication capacity was measured on the Communication Capacity Scale. Palliative sedation therapy succeeded in symptom alleviation in 83% of the cases. Median time elapsed before patients initially had one continuous hour of deep sedation was 60 minutes, but 49% of the patients awakened once after falling into a deeply sedated state. The percentage of patients who were capable of explicit communication decreased from 40% before sedation to 7.1% 4 hours after sedation, and the mean Communication Capacity Score significantly decreased to the level of 15 points (P < 0.001). The respiratory rates did not significantly decrease after sedation (18 +/- 9.0 to 16 +/- 9.4/min, P = 0.62), but respiratory and/or circulatory suppression (respiratory rate or=4 on 0-10 scales) with walking ability, normal work, sleep, enjoyment of life, mood, and general activity. Moderate to severe symptom levels of anxiety and depression (HADS-A and HADS-D scores >or=11 on 0-21 scales) occurred in 35% and 28% of patients, respectively. Patients reported greater sleep problems compared with the general U.S. population and significant impairment in both physical and mental functioning (SF-12v2) compared with subjects with diabetes. The mean EQ-5D utility score was 0.5 +/- 0.3. Greater pain levels in DPN (mild to moderate to severe) corresponded with higher symptom levels of anxiety and depression, more sleep problems, and lower utility ratings and physical and mental functioning, (all Ps < 0.01). Painful DPN is associated with decrements in many aspects of patients' lives: physical and emotional functioning, affective symptoms, and sleep problems. The negative impact is higher in patients with greater pain severity. PMID- 16256905 TI - Hugh Hampton Young, benign prostatic hyperplasia, and "the cure of prostatic obstruction". PMID- 16256906 TI - Prognostic factors and longterm survival after hepatic resection for hepatocellular carcinoma originating from noncirrhotic liver. AB - BACKGROUND: The incidence of hepatocellular carcinoma (HCC) in cirrhotic and noncirrhotic liver is increasing in the world, probably because of the high prevalence of infections by hepatitis B and C viruses. Despite numerous publications on hepatic resection, prognostic factors for intrahepatic recurrence and survival are not well known for patients with HCC without cirrhosis. STUDY DESIGN: One hundred eight consecutive patients with HCC in noncirrhotic liver have been treated by hepatic resection in the past 18 years in our center. Clinical, biologic, and histopathologic parameters of these patients were collected. Risk factors for intrahepatic recurrence and prognostic factors for survival were evaluated by univariate and multivariate analyses. RESULTS: Postoperative morbidity and mortality rates were 23% and 6.5%, respectively. The 3- and 5-year disease-free and overall survival rates were 55% and 43%, and 43% and 29%, respectively. Blood transfusion, absence of tumor capsule, and daughter nodules were independently associated with overall survival. But the only risk factors for recurrence were blood transfusion, absence of tumor capsule, daughter nodules, and margin resection < 10 mm. CONCLUSIONS: In the treatment of HCC without cirrhosis, hepatectomy remains a safe and legitimate treatment, but longterm results are impaired by a high rate of early recurrence likely related to metastatic dissemination. Only histopathologic factors related to the tumor are predictive of recurrence and overall survival. PMID- 16256907 TI - Resection of hepatocellular carcinoma in noncirrhotic liver: analysis of risk factors for survival. AB - BACKGROUND: The aim of this study was to identify factors predictive of survival after curative resection of hepatocellular carcinoma (HCC) in noncirrhotic liver. STUDY DESIGN: Eighty-four patients underwent resection of HCC in noncirrhotic liver between January 1998 and December 2003. Univariate and multivariable analyses were used to retrospectively identify factors associated with overall survival and disease-free survival when resection was curative for the primary tumor. RESULTS: Overall 1-, 3-, and 5-year survival rates were 77.8%, 55.0%, and 44.4%, respectively, and 84.0%, 62.0%, and 50.0% when resection was curative for the primary tumor. HCC recurred in 27 patients (39.1%). Recurrence was intrahepatic in 14 patients (51.9%), extrahepatic in 3 patients (11.1%), and both intra- and extrahepatic in the remaining 10 patients (37.0%). In multivariable analysis, three independent factors were associated with poorer overall survival and recurrence-free survival, namely multiple tumors, macroscopic vascular invasion, and nonuse of adjuvant iodine-131-iodized oil. CONCLUSIONS: Aggressive operation is an effective treatment for HCC in noncirrhotic patients, whatever the degree of liver fibrosis. Multiple tumors and macroscopic vascular invasion are poor prognostic factors. Postoperative iodine-131-iodized oil injection appears to prevent recurrence and improve overall survival, although this needs to be confirmed in a prospective randomized trial. PMID- 16256908 TI - Hepatic resection for primary or secondary malignancies with involvement of the inferior vena cava: is this operation safe or hazardous? AB - BACKGROUND: This study evaluated surgical techniques and results of patients with tumors who had undergone liver resection with partial resection and reconstruction of the IVC. STUDY DESIGN: We performed a retrospective analysis of all patients who underwent combined liver and IVC resection and reconstruction at a single institution. We identified 19 patients and two categories of tumors, primary (n = 8) and metastatic (n = 11). In 12 patients, a direct suture of the IVC was performed; in 3 patients a pericardium bovine patch was applied; in another 4 patients the IVC was replaced by PTFEt prosthesis. In nine patients, total hepatic vascular occlusion was required. RESULTS: Perioperative mortality was 5.9%, related to technical complications and hepatic insufficiency. Postoperative morbidity was 57.9%. Median survival time was 32 months (range 3 to 125 months). The 1-, 2-, and 5-year cumulative survival rates were 78.9%, 68%, and 49.1%, respectively. Tumor recurrence appeared in 13 patients and was the main cause of death (55.5%). Among the seven patients suffering from hepatocellular carcinoma, three are still alive at 31, 60, and 125 months after resection. In this group, 1-, 2-, and 5-year survival rates were 71.4%, 57.1%, and 38.1%. Among the 11 patients resected for colorectal liver metastases, the 1 , 2-, and 5-year survival rates were 81.8%, 62.3%, and 51.9%, respectively. CONCLUSIONS: Liver resection combined with IVC resection and reconstruction is a feasible procedure that can be performed with an acceptable operative risk leading to longterm outcome in selected patients. PMID- 16256909 TI - Factors associated with insulin and narcotic independence after islet autotransplantation in patients with severe chronic pancreatitis. AB - BACKGROUND: For patients who suffer from severe chronic pancreatitis, total pancreatectomy can alleviate pain, and islet autotransplantation (IAT) might preserve endocrine function and circumvent the complications of diabetes. Factors that determine success after this operation have not been clearly defined. STUDY DESIGN: From 2000 to 2004, 45 total or subtotal pancreatectomies with IAT were performed. Patient characteristics, narcotic usage and insulin requirements were recorded at routine followup. Narcotic usage was standardized by conversion to morphine equivalents (MEs). Univariate and multivariate statistical analyses were performed to determine factors associated with insulin and narcotic independence. RESULTS: Forty-five patients (30 women, 15 men), with a mean age of 39 years (range 16 to 62 years) underwent total or completion (n=41) or subtotal (n=4) pancreatectomies with IAT. Forty percent of patients were insulin free after a mean followup of 18months (range 1 to 46months). Factors associated in univariate analyses with insulin independence included female gender (p=0.004), lower body weight (kg) (p=0.04), more islet equivalents per kg body weight (IEQ/kg) transfused (<0.05), lower mean insulin requirement for the first 24hours postoperation (p=0.002), and lower mean insulin requirement at discharge (p=0.0005). A multiple logistic regression using gender, body mass index, and IEQ/kg identified female gender as the only notable variable associated with insulin independence. There was a notable reduction (p < 0.0001) of postoperative MEs (mean 90 mg) compared with preoperative MEs (mean 206 mg) for the entire cohort; 58% of patients are narcotic independent. In the subset of patients with>5months followup (n=32), 23 (72%) are narcotic free, with a substantial decrease in ME usage (p=0.01). CONCLUSIONS: The likelihood of glycemic control after IAT is related to both patient characteristics and islet cell mass. Based on these data, more islet cells may be required for insulin independence than previously thought. PMID- 16256910 TI - Tamoxifen use in patients with ductal carcinoma in situ and T1a/b N0 invasive carcinoma. AB - BACKGROUND: The purpose of this study was to determine how often patients with ductal carcinoma in situ and T1a/b N0 cancer are offered and accept tamoxifen for secondary chemoprevention. STUDY DESIGN: A retrospective review of 284 patients with T1a/b N0 invasive cancer treated between February 1995 and December 2001 and 129 patients with DCIS treated after September 1998 was carried out. Patient and tumor characteristics associated with being offered and accepting tamoxifen were compared. RESULTS: Tamoxifen was offered to 67% of the invasive cancer patients and accepted by 76% (51% of the entire group). Hormone receptor status was the only significant predictor of being offered tamoxifen (p = 0.004). Older age (p = 0.04), Caucasian race (p = 0.01), and parity (p = 0.04) in premenopausal women were significant predictors of tamoxifen acceptance on univariate analysis. After the publication of the National Surgical Adjuvant Breast and Bowel Project P-1 trial, significantly more patients were offered tamoxifen (p = 0.02), but acceptance rates did not change. Tamoxifen was offered to 91% of the ductal carcinoma in situ patients and accepted by 73% (67% overall). Lumpectomy was associated with significantly higher rates of being offered (p = 0.02) and accepting tamoxifen (p = 0.002) on univariate analysis. CONCLUSIONS: Factors associated with tamoxifen risks and benefits correlate poorly with the use of the drug. PMID- 16256911 TI - Percutaneous transesophageal gastrostomy tube for decompression of malignant obstruction: report of the first case and our series in the US. AB - BACKGROUND: Historically, surgical gastrostomies, gastrojejunostomy, and percutaneous endoscopic gastrostomy have been used palliatively. Recently, enteral stenting has also provided a means of reestablishing gastrointestinal flow in proximal and colonic obstructions. STUDY DESIGN: Seven patients with known intraabdominal malignancy leading to gastrointestinal obstruction were evaluated for PTEG. Ultrasonography, fluoroscopy, and a rupture-free balloon were used in placement. An endoscope was not used. Consent was obtained from all patients. The procedure was performed by a single surgical endoscopist in an endoscopy suite. Patients had appropriate hemodynamic monitoring with pulse oximetry, and they were given preprocedure antibiotics and sedation. RESULTS: PTEG was successfully placed and alleviated symptoms in all seven patients. One complication occurred; in the fourth patient subcutaneous emphysema developed on postoperative day 1, and was managed nonoperatively. All patients were discharged from the hospital. CONCLUSIONS: PTEG is a safe and effective technique for decompression in malignant gastrointestinal obstruction. PMID- 16256912 TI - Prospective evaluation of a screening protocol to exclude deep vein thrombosis on the basis of a combination of quantitative D-dimer testing and pretest clinical probability score. AB - BACKGROUND: Clinical signs and symptoms such as swelling, pain, and redness are unreliable markers of deep vein thrombosis (DVT). Because of this venous duplex scanning (VDS) has been heavily used in DVT detection. The purpose of this study was to determine if a combination of D-dimer testing and pretest clinical score could reduce the use of VDS in symptomatic patients with suspected DVT. STUDY DESIGN: One hundred seventy-four consecutive patients with suspected DVT were prospectively evaluated using pretest clinical probability (PCP) score and D dimer testing before VDS. After calculating clinical probability scores developed by Wells and associates, patients were divided into low risk (or=3 points) PCP. RESULTS: One hundred fifty-eight patients were enrolled. The prevalence of DVT in this study was 37%. Thirty-eight patients (24%) were classified as low risk, 64 (41%) as moderate risk, and 56 (35%) as high risk PCP. DVT was identified in only one patient (2.6%) with low risk PCP. In contrast, DVT was found in 22 (34%) with moderate risk, and 35 (63%) with high risk PCP. In the high and moderate risk PCP groups, positive scan patients had a markedly higher value of D-dimer assay than negative scan patients (p=0.0001 and p=0.0057, respectively). In the low risk PCP patients, D-dimer testing provided 100% sensitivity, 46% specificity, 4.8% positive predictive value, and 100% negative predictive value in the diagnosis of DVT. Similarly, in the moderate risk PCP, the D-dimer testing showed 100% sensitivity, 45% specificity, 49% positive predictive value, and 100% negative predictive value. In the high risk group, D-dimer testing achieved 100% sensitivity, 57% specificity, 80% positive predictive value, and 100% negative predictive value in the diagnosis of DVT. These results suggested that 36 of 158 patients who had a non-high PCP (low and moderate PCP) and a normal D-dimer concentration were considered to have no additional investigation, so VDS could have been reduced by 23% (36/158). CONCLUSIONS: A combination of D-dimer testing and clinical probability score may be effective in avoiding unnecessary VDS in suspected symptomatic DVT in the low and moderate PCP patients. The need for VDS could be reduced by 23% despite a relatively high prevalence of DVT. PMID- 16256914 TI - A multidisciplinary youth violence-prevention initiative: impact on attitudes. AB - BACKGROUND: In a previous report, enhanced resource commitment at a Level I trauma center was associated with improved outcomes for most major categories of injured patients, except those with gunshot wounds, which disproportionately affected the young (ages 15 to 24 years). We hypothesized that a primary violence prevention initiative geared toward changing attitudes about interpersonal conflict among at-risk youths can be effective. STUDY DESIGN: Between May 2002 and November 2003, 97 youths (mean age 12.6 years) were recruited from one of two Police Athletic League centers in the catchment area of our Level I trauma center. Participant attitudes about interpersonal conflicts were surveyed with six previously validated scales before and after a hospital tour with a video and slide presentation graphically depicting the results of gun violence. Mean differences in scores between pre- and postintervention surveys were assessed. RESULTS: Of the 97 participants, 48 (49.4%) completed the intervention program with both the pre- and postintervention tests, with a mean of 25.8 days between tests. There was a statistically significant reduction in the Beliefs Supporting Aggression scale (mean -0.38 U; 95% CI, -0.23 to -0.54; p < 0.01), and a trend toward reduced Likelihood of Violence (mean -0.17 U; 95% CI, 0.01 to -0.34; p = 0.06). CONCLUSIONS: A multidisciplinary violence-prevention outreach program can produce short-term improvement in beliefs supporting aggression among at-risk youth. Longterm impact of this attitude change needs to be examined in future studies. PMID- 16256913 TI - Treatment of bile acid malabsorption using ileal stem cell transplantation. AB - BACKGROUND: We hypothesized that ileal stem cell clusters transplanted into a segment of jejunum can be used to treat bile acid malabsorption. STUDY DESIGN: In adult Lewis rats, a 15-cm segment of jejunum was isolated with its blood circulation left intact and partially stripped of enterocytes using luminal high velocity perfusions with 3mmol/L ethylenediamine tetra-acetic acid solutions. Continuity was restored by anastomosing the proximal and distal gut. Ileal stem cell clusters were harvested from neonatal Lewis rats and transplanted into the stripped segments to generate a "neoileum." After 4weeks, recipients underwent resection of the native ileum, and the isolated neoileum was anastomosed in its place. After an additional 4weeks, a 48-hour stool collection was performed. The engrafted segment was harvested for taurocholate uptake studies, ileal bile acid transporter (IBAT) protein by immunohistomorphometry, and IBAT mRNA quantitation by reverse transcription polymerease chain reaction. Data were analyzed by ANOVA/t-test. Rats undergoing ileectomy, jejunectomy, or sham operations served as controls. RESULTS: Total bile acid loss in the stool was markedly lower in rats with a neoileum compared with rats with an ileectomy (p < 0.001). Total taurocholate uptake was notably increased in the neoileum compared with the jejunum (p < 0.001). IBAT protein signal intensity was considerably higher in the neoileum compared with jejunum (p < 0.001). IBAT mRNA amounts in the neoileal group were comparable with those in normal rat ileum and were considerably higher (p = 0.003) than in the jejunum. CONCLUSIONS: Ileal stem cell clusters were used to establish a new zone of bile acid uptake and IBAT expression in a jejunal segment. This neoileum eliminated loss of bile acids in the stool after ileectomy. This is the first time that transplantation of intestinal stem cell clusters has been shown to correct a clinical malabsorption syndrome. PMID- 16256915 TI - Do resident duty hour restrictions reduce technical complications of emergency laparoscopic cholecystectomy? AB - BACKGROUND: The impact of resident duty hour restrictions on patient care has not been assessed. STUDY DESIGN: We studied 275 patients undergoing emergency cholecystectomy before and after duty hour regulations instituted by the Accreditation Council for Graduate Medical Education. Operations were stratified into 6-hour intervals from the time in-hospital call began. Procedure-related complications (bile duct injury, cystic duct leak, abdominal hemorrhage, trocar injury, intraabdominal/wound infection, unrecognized retained stone) were the primary outcomes variables. RESULTS: Complications occurred after 7 of 107 (6.5%) operations performed before duty hour restrictions, which was not different from 15 of 168 (8.9%) after duty hour restrictions. In both periods, all complications followed operations that began within the first 18 hours of duty. Patients with complications had longer operative times (p = 0.038) and a higher proportion of operations lasting 120 minutes or longer (p = 0.006). Comparing patients with and without complications, there were no significant differences in patient demographics, operative complexity, or PGY level of the surgeon. Only operative time of 120 minutes or longer retained significance in the multivariable model (p = 0.0023; odds ratio, 4.05; 95% CI, 1.65-9.97). CONCLUSIONS: There was no correlation between imposition of duty hour restrictions and technical complication rates in this study. Duration of operative time of 120 minutes or longer was the only independent marker, suggesting that technical complications are a function of operative complexity, not duration of duty. These data suggest that duty hour restrictions might not have a measurable influence on the surgical complication rate after emergency cholecystectomy. PMID- 16256916 TI - Comparison of urban and rural general surgeons: motivations for practice location, practice patterns, and education requirements. AB - BACKGROUND: The purpose of this study is to determine the differences between rural and urban surgeons with regard to practice patterns, factors in choosing a practice location, and educational needs. STUDY DESIGN: A list of surgeons obtained from the American Medical Association was examined using the Office of Management and Budget definition of rural. Seventeen hundred rural surgeons were mailed surveys; 421 responded. One hundred fourteen urban surgeons were contacted by telephone. Questions were designed to measure job and community satisfaction, factors influencing their decision to practice in their current location, spectrum and volume of cases, and their perceived educational needs. RESULTS: Age distribution did not differ markedly between urban and rural surgeons. Motivation to practice in their current location varied considerably between urban and rural surgeons. Both groups equally rated quality of life as the leading factor influencing their current practice location. Urban surgeons rated other factors, such as income, practice growth, hospital facilities, and proximity to family, higher than rural surgeons. Practice patterns and educational needs also varied between the two groups. Rural surgeons performed more procedures per year with more variety in procedure type. Both groups felt that additional training in advanced laparoscopic techniques would be helpful, and rural surgeons felt that additional training in the surgical subspecialty areas was important. CONCLUSIONS: Although rural and urban surgeons do not differ in age or the importance of lifestyle in deciding career location, different factors do impact their choice of location. Practice pattern and educational needs varied markedly between rural and urban general surgeons. PMID- 16256917 TI - Paint-only is equivalent to scrub-and-paint in preoperative preparation of abdominal surgery sites. AB - BACKGROUND: Antiseptic preoperative skin site preparation is used to prepare the operative site before making a surgical incision. The goal of this preparation is a reduction in postoperative wound infection. The most straightforward technique necessary to achieve this goal remains controversial. STUDY DESIGN: A prospective randomized trial was designed to prove equivalency for two commonly used techniques of surgical skin site preparation. Two hundred thirty-four patients undergoing nonlaparoscopic abdominal operations were consented for the trial. Exclusion criteria included presence of active infection at the time of operation, neutropenia, history of skin reaction to iodine, or anticipated insertion of prosthetic material at the time of operation. Patients were randomized to receive either a vigorous 5-minute scrub with povidone-iodine soap, followed by absorption with a sterile towel, and a paint with aqueous povidone iodine or surgical site preparation with a povidone-iodine paint only. The primary end point of the study was wound infection rate at 30 days, defined as presence of clinical signs of infection requiring therapeutic intervention. RESULTS: Patients randomized to the scrub-and-paint arm (n = 115) and the paint only arm (n = 119) matched at baseline with respect to age, comorbidity, wound classification, mean operative time, placement of drains, prophylactic antibiotic use, and surgical procedure (all p > 0.09). Wound infection occurred in 12 (10%) scrub-and-paint patients, and 12 (10%) paint-only patients. Based on our predefined equivalency parameters, we conclude equivalence of infection rates between the two preparations. CONCLUSIONS: Preoperative preparation of the abdomen with a scrub with povidone-iodine soap followed by a paint with aqueous povidone-iodine can be abandoned in favor of a paint with aqueous povidone-iodine alone. This change will result in reductions in operative times and costs. PMID- 16256918 TI - What's new in ophthalmic surgery. PMID- 16256919 TI - What's new in general surgery: endocrine surgery. PMID- 16256920 TI - Using a comprehensive examination to assess multiple competencies in surgical residents: does the oral examination still have a role? AB - BACKGROUND: While specialty-level evaluations evolve from traditional examinations to objective structured clinical examination-like assessments, a broader range of competencies are tested; consequently, examiners are forced to integrate results when making a determination of competency. The aim of this study was to describe how experts weigh relative performances on specific components of a comprehensive examination to make decisions of overall competency. STUDY DESIGN: The Patient Assessment and Management Examination is a standardized patient-based assessment of general surgery in which each 25-minute station encompasses four components: history and physical examination, investigation interpretation, diagnosis and treatment discussion with the patient, and a structured oral examination (SOE). A six-station Patient Assessment and Management Examination was administered to 21 senior surgery residents. Surgeons marked each station with global rating scales and, in addition, provided an end-of-station overall global assessment of performance. A "gold-standard" examination pass-or-fail decision was determined through videotape review of each candidate's performance across six stations by two blinded surgeons. Multiple linear regression analysis was used to determine which components were associated with the end-of-station overall global assessments. Multivariable logistic regression was used to determine which components were associated with the final "gold-standard" pass-or-fail assessment. RESULTS: The only component notably (p < 0.005) associated with end-of-station global assessment for all six stations was the SOE. Mean SOE score was the only notable independent variable associated with the gold-standard pass-or-fail decision (R(2) = 0.63, p < 0.001). CONCLUSIONS: Performance on the SOE section of a multicompetency examination is markedly associated with the final determination of competency. These results have implications for the design and implementation of comprehensive specialty-level assessments. PMID- 16256921 TI - Complications of construction and closure of temporary loop ileostomy. PMID- 16256922 TI - The liver and hemorrhagic shock. PMID- 16256923 TI - Development of pituitary surgery: the Chicago contributions. PMID- 16256924 TI - Postraumatic innominate artery pseudoaneurysm. PMID- 16256925 TI - Superior mesenteric artery syndrome. PMID- 16256926 TI - Technical refinements in the supracricoid partial laryngectomy to optimize functional outcomes. PMID- 16256927 TI - Intraoperative frozen-section evaluation can reduce accuracy of pathologic assessment of sentinel nodes in melanoma patients. PMID- 16256929 TI - Roux-en-Y gastric bypass. PMID- 16256930 TI - Edler resuscitated? PMID- 16256932 TI - Driving speed and the risk of road crashes: a review. AB - Driving speed is an important factor in road safety. Speed not only affects the severity of a crash, but is also related to the risk of being involved in a crash. This paper discusses the most important empirical studies into speed and crash rate with an emphasis on the more recent studies. The majority of these studies looked at absolute speed, either at individual vehicle level or at road section level. Respectively, they found evidence for an exponential function and a power function between speed and crash rate. Both types of studies found evidence that crash rate increases faster with an increase in speed on minor roads than on major roads. At a more detailed level, lane width, junction density, and traffic flow were found to interact with the speed-crash rate relation. Other studies looked at speed dispersion and found evidence that this is also an important factor in determining crash rate. Larger differences in speed between vehicles are related to a higher crash rate. Without exception, a vehicle that moved (much) faster than other traffic around it, had a higher crash rate. With regard to the rate of a (much) slower moving vehicle, the evidence is inconclusive. PMID- 16256933 TI - Isometamidium sensitivity of Trypanosoma congolense stocks from cattle in West Africa tested in mice and the drug incubation infectivity test. AB - Four Trypanosoma (T.) congolense reference clones with known isometamidium sensitivity and 16 T. congolense stocks from cattle in Kenedougou in south western Burkina Faso, an area with known history of drug resistance, were characterised with the standard mouse test (SMT) and the drug incubation infectivity test (DIIT). All field stocks from Kenedougou were resistant to 1.0 mg/kg bw isometamidium in the SMT. Fourteen stocks (87.5%) also proved to be refractory to 10 mg/kg bw. Testing with the DIIT confirmed the results of the SMT. By comparison to reference clones, all the Kenedougou populations expressed high levels of resistance to isometamidium. PMID- 16256934 TI - The nature of expertise: a review. AB - This paper reviews the complex topic of expertise. It begins with an explanation for the range of interest and viewpoints, and moves on to a discussion of the nature and study of expertise. The diversity in definitions, domains, disciplines, and the impact of these factors on approaches to investigation, are offered as possible explanations for some of the differences that appear to run through the literature. Specific attention is given to issues that may concern ergonomists interested in understanding expertise from the perspective of work in complex and dynamic settings. We suggest a move away from traditional novice expert experimental designs to study of work in a naturalistic way, studying tasks and activities that are sufficiently challenging so that real expertise can be elicited. PMID- 16256935 TI - Gangliosides activate the phosphatase activity of the erythrocyte plasma membrane Ca2+-ATPase. AB - The previous studies showed that gangliosides modulated the ATPase activity of the PMCA from porcine brain synaptosomes [Yongfang Zhao, Xiaoxuan Fan, Fuyu Yang, Xujia Zhang, Arch. Biochem. Biophys. 427 (2004) 204-212]. The effects of gangliosides on the hydrolysis of p-nitrophenyl phosphate (pNPP) catalyzed by the erythrocyte plasma membrane Ca(2+)-ATPase, which was characterized as E(2) conformer of the enzyme, were studied. The results showed that pNPPase activity was stimulated up to seven-fold, depending upon the different gangliosides used with GD1b>GM1>GM2>GM3 approximately Asialo-GM1. Under the same conditions, the ATPase activity was also activated, suggesting that gangliosides should modify both E(1) and E(2) conformer of the enzyme. The Ca(2+), which drove the enzyme to E(1) conformation, inhibited the pNPPase activity, but with the similar half maximal inhibitory concentrations (IC(50)) in the presence and the absence of gangliosides. Moreover, the pNPPase activity was also inhibited by the raise in ATP concentrations. Gangliosides caused a large increase in V(max), but had no effect on the apparent affinity (K(m)) of the enzyme for pNPP. The kinetic analysis indicated that gangliosides could modulate the erythrocyte PMCA through stabilizing E(2) conformer. PMID- 16256936 TI - Glucose-6-phosphate dehydrogenase regulation during hypometabolism. AB - Glucose-6-phosphate dehydrogenase (G6PDH) from hepatopancreas of the land snail, Otala lactea, shows distinct changes in properties between active and estivating (dormant) states, providing the first evidence of pentose phosphate cycle regulation during hypometabolism. Compared with active snails, G6PDH Vmax increased by 50%, Km for glucose-6-phosphate decreased by 50%, Ka Mg x citrate decreased by 35%, and activation energy (from Arrhenius plots) decreased by 35% during estivation. DEAE-Sephadex chromatography separated two peaks of activity and in vitro incubations stimulating protein kinases or phosphatases showed that peak I (low phosphate) G6PDH was higher in active snails (57% of activity) whereas peak II (high phosphate) G6PDH dominated during estivation (71% of total). Kinetic properties of peaks I and II forms mirrored the enzyme from active and estivated states, respectively. Peak II G6PDH also showed reduced sensitivity to urea inhibition of activity and greater stability to thermolysin protease treatment. The interconversion of G6PDH between active and estivating forms was linked to protein kinase G and protein phosphatase 1. Estivation induced phosphorylation of G6PDH may enhance relative carbon flow through the pentose phosphate cycle, compared with glycolysis, to help maintain NADPH production for use in antioxidant defense. PMID- 16256937 TI - Investigation of CC and CXC chemokine quaternary state mutants. AB - The chemokine family forms two different types of homodimer despite members sharing nearly identical folds. To study the formation of quaternary structure in this family, rational mutagenesis was employed on a representative member of each subfamily (MIP-1beta and IL-8). The variants were studied by analytical ultracentrifugation and NMR, and it was determined that formation of a folded monomer from a natural chemokine dimer is reasonably facile, while conversion between dimer types is not. Monomeric variants of MIP-1beta and IL-8 were randomly mutated and a lambda phage-based selection system was employed in a novel way to screen for dimerization. A total of 6,000,000 random mutants were screened, but no dimers were formed, suggesting again that the chemokine fold is robust and amenable to sequence variation, while the chemokine dimer is much more difficult to attain. This work represents a biophysical analysis of an array of chemokine quaternary state variants. PMID- 16256938 TI - Evidence for an indirect transcriptional regulation of glucose-6-phosphatase gene expression by liver X receptors. AB - Liver X receptor (LXR) paralogues alpha and beta (LXRalpha and LXRbeta) are members of the nuclear hormone receptor family and have oxysterols as endogenous ligands. LXR activation reduces hepatic glucose production in vivo through the inhibition of transcription of the key gluconeogenic enzymes phosphoenolpyruvate carboxykinase and glucose-6-phosphatase (G6Pase). In the present study, we investigated the molecular mechanisms involved in the regulation of G6Pase gene expression by LXR. Both T0901317, a synthetic LXR agonist, and the adenoviral overexpression of either LXRalpha or LXRbeta suppressed G6Pase gene expression in H4IIE hepatoma cells. However, compared to the suppression of G6Pase expression seen by insulin, the decrease of G6Pase mRNA by LXR activation was delayed and was blocked by cycloheximide, an inhibitor of protein synthesis. These observations, together with the absence of a conserved LXR-binding element within the G6Pase promoter, suggest an indirect inhibition of G6Pase gene expression by liver X receptors. PMID- 16256939 TI - Targeted inhibition of alphavbeta3 integrin with an RNA aptamer impairs endothelial cell growth and survival. AB - Alphavbeta3 integrin is a crucial factor involved in a variety of physiological processes, such as cell growth and migration, tumor invasion and metastasis, angiogenesis, and wound healing. Alphavbeta3 integrin exerts its effect by regulating endothelial cell (EC) migration, proliferation, and survival. Inhibiting the function of alphavbeta3 integrin, therefore, represents a potential anti-cancer, anti-thrombotic, and anti-inflammatory strategy. In this study, we tested an RNA aptamer, Apt-alphavbeta3 that binds recombinant alphavbeta3 integrin, for its ability to bind endogenous alphavbeta3 integrin on the surface of cells in culture and to subsequently affect cellular response. Our data illustrate that Apt-alphavbeta3 binds alphavbeta3 integrin expressed on the surface of live HUVECs. This interaction significantly decreases both basal and PDGF-induced cell proliferation as well as inhibition of cell adhesion. Apt alphavbeta3 can also reduce PDGF-stimulated tube formation and increase HUVEC apoptosis through inhibition of FAK phosphorylation pathway. Our results demonstrate that by binding to its target, Apt-alphavbeta3 can efficiently inhibit human EC proliferation and survival, resulting in reduced angiogenesis. It predicts that Apt-alphavbeta3 could become useful in both tumor imaging and the treatment of tumor growth, atherosclerosis, thrombosis, and inflammation. PMID- 16256940 TI - Bidirectional regulations for glutamate and GABA release in the hippocampus by alpha7 and non-alpha7 ACh receptors. AB - In the assay of glutamate and gamma-aminobutyric acid (GABA) with a high performance liquid chromatography, spontaneous release of glutamate and GABA from rat hippocampal slices was significantly enhanced by mecamylamine, an inhibitor of non-alpha7 ACh receptors, or alpha-bungarotoxin, an inhibitor of alpha7 ACh receptors in the absence of tetrodotoxin (TTX), but not in the presence of TTX. Nicotine significantly enhanced glutamate and GABA release in the absence of TTX, that is abolished by mecamylamine or alpha-bungarotoxin, while it had no effect on the release in the presence of TTX. In the recording of alpha-amino-3-hydroxy 5-methyl-4-isoxazole propionic acid (AMPA) receptor-mediated excitatory postsynaptic currents (AMPA-EPSCs) and GABA(A) receptor-mediated inhibitory postsynaptic currents (GABA(A)-IPSCs) from CA1 pyramidal neurons of rat hippocampal slices, nicotine did not affect the rate and amplitude of AMPA-EPSCs and AMPA-miniature EPSCs. In contrast, nicotine significantly increased the rate of GABA(A)-IPSCs, without affecting the amplitude, but such effect was not obtained with GABA(A)-miniature IPSCs. The collective results suggest that alpha7 and non-alpha7 ACh receptors expressed in the hippocampus, activated under the basal conditions, inhibit release of glutamate and GABA controlled through multi synaptic relays, but that otherwise, those receptors, highly activated by nicotine, stimulate both the release, with a part of GABA released from interneurons transmitting to CA1 pyramidal neurons. Furthermore, the results also suggest that alpha7 and non-alpha7 ACh receptors do not have potency sufficiently to modulate glutamate and GABA release controlled by single synapses. PMID- 16256941 TI - Overexpression of calmodulin induces cardiac hypertrophy by a calcineurin dependent pathway. AB - The possible role of calcineurin in cardiac hypertrophy induced by calmodulin (CaM) overexpression in the heart was investigated. CaM transgenic (CaM-TG) mice developed marked cardiac hypertrophy and exhibited up-regulation of atrial natriuretic factor (ANF) and beta-myosin heavy chain gene expression in the heart during the first 2 weeks after birth. The activity of calcineurin in the heart was also significantly increased in CaM-TG mice compared with wild-type littermates. Treatment of CaM-TG mice with the calcineurin inhibitor FK506 (1mg/kg per day) prevented the increase in the heart-to-body weight ratio as well as that in cardiomyocyte width. FK506 also inhibited the induction of fetal-type cardiac gene expression in CaM-TG mice. Overexpression of CaM in cultured rat cardiomyocytes activated the ANF gene promoter in a manner sensitive to FK506. Activation of a calcineurin-dependent pathway thus contributes to the development of cardiac hypertrophy induced by CaM overexpression in the heart. PMID- 16256942 TI - Human cytochrome c enters murine J774 cells and causes G1 and G2/M cell cycle arrest and induction of apoptosis. AB - Cytochrome c is well known as a carrier of electrons during respiration. Current evidence indicates that cytochrome c also functions as a major component of apoptosomes to induce apoptosis in eukaryotic cells as well as an antioxidant. More recently, a prokaryotic cytochrome c, cytochrome c(551) from Pseudomonas aeruginosa, has been shown to enter in mammalian cells such as the murine macrophage-like J774 cells and causes inhibition of cell cycle progression. Much less is known about such functions by mammalian cytochromes c, particularly the human cytochrome c. We now report that similar to P. aeruginosa cytochrome c(551), the purified human cytochrome c protein can enter J774 cells and induce cell cycle arrest at the G(1) to S phase, as well as at the G(2)/M phase at higher concentrations. Unlike P. aeruginosa cytochrome c(551) which had no effect on the induction of apoptosis, human cytochrome c induces significant apoptosis and cell death in J774 cells, presumably through inhibition of the cell cycle at the G(2)/M phase. When incubated with human breast cancer MCF-7 and normal mammary epithelial cell line MCF-10A1 cells, human cytochrome c entered in both types of cells but induced cell death only in the normal MCF-10A1 cells. The ability of human cytochrome c to enter J774 cells was greatly reduced at 4 degrees C, suggesting energy requirement in the entry process. PMID- 16256943 TI - Depletion of vesicle-tethering factor p115 causes mini-stacked Golgi fragments with delayed protein transport. AB - Depletion of p115 with small interfering RNA caused fragmentation of the Golgi apparatus, resulting in dispersed distribution of stacked short cisternae and a vesicular structure (mini-stacked Golgi). The mini-stacked Golgi with cis- and trans-organization is functional in protein transport and glycosylation, although secretion is considerably retarded in p115 knockdown cells. The fragmented Golgi was further disrupted by treatment with breferdin A and reassembled into the mini stacked Golgi by removal of the drug, as observed in control cells. In addition, p115 knockdown cells maintained retrograde transport from the Golgi to the endoplasmic reticulum, although the rate was not as efficient as in control cells. While no alternation of microtubule networks was found in p115 knockdown cells, the fragmented Golgi resembled those in cells treated with anti microtubule drugs. The results suggest that p115 is involved in vesicular transport between endoplasmic reticulum and the Golgi, along with microtubule networks. PMID- 16256944 TI - siRNA-mediated inhibition of endogenous Huntington disease gene expression induces an aberrant configuration of the ER network in vitro. AB - Huntingtin is a ubiquitously expressed cytoplasmic protein encoded by the Huntington disease (HD) gene, in which a CAG expansion induces an autosomal dominant progressive neurodegenerative disorder; however, its biological function has not been completely elucidated. Here, we report for the first time that short interfering RNA (siRNA)-mediated inhibition of endogenous Hdh (a mouse homologue of huntingtin) gene expression induced an aberrant configuration of the endoplasmic reticulum (ER) network in vitro. Studies using immunofluorescence microscopy with several ER markers revealed that the ER network appeared to be congregated in various types of cell lines transfected with siRNA directed against Hdh, but not with other siRNAs so far tested. Other subcellular organelles and structures, including the nucleus, Golgi apparatus, mitochondria, lysosomes, microtubules, actin cytoskeletons, cytoplasm, lipid rafts, and plasma membrane, exhibited normal configurations. Western blot analysis of cellular prion protein (PrP(C)) revealed normal glycosylation, which is a simple marker of post-translational modification in the ER and Golgi compartments, and immunofluorescence microscopy detected no altered subcellular distribution of PrP(C) in the post-ER compartments. Further investigation is required to determine whether the distorted ER network, i.e., loss of the huntingtin function, participates in the development of HD. PMID- 16256945 TI - Development of novel fluorescent probe 3-perylene diphenylphosphine for determination of lipid hydroperoxide with fluorescent image analysis. AB - A novel fluorescent probe 3-perylene diphenylphosphine (3-PeDPP) was synthesized for the direct analysis of lipid hydroperoxides. The structure of 3-PeDPP was identified by the spectroscopic data, FAB-MS, (1)H NMR, and (13)C NMR. The reactivities of 3-PeDPP with lipid hydroperoxides were investigated in chloroform/MeOH homogeneous solutions and PC liposome model systems oxidized by either 2,2'-azobis(2-amidinopropane)dihydrochloride and photosensitized oxidation. The fluorescence intensity derived from 3-perylene diphenylphosphineoxide (3-PeDPPO) increased proportionally with amount of hydroperoxides produced in homogeneous solutions and liposome model systems. 3 PeDPP was easily incorporated into mouse myeloma SP2 cells and thin tissue section for dynamic membrane lipid peroxidation studies. Linear correlations between fluorescence intensity and amount of hydroperoxides in the cell membrane and tissue sections were obtained. The fluorescence intensity from 2-dimensional image analysis was also well correlated with lipid hydroperoxide level in these models. Thus, the novel probe 3-PeDPP is useful for the direct determination of lipid hydroperoxides in biological materials. PMID- 16256946 TI - Effect of serum on the down-regulation of CHOP-10 during differentiation of 3T3 L1 preadipocytes. AB - Hormonal induction of growth-arrested 3T3-L1 preadipocytes in medium containing fetal bovine serum (FBS) triggers a signaling cascade that culminates in adipogenesis. The transcription factor, C/EBPbeta, is expressed early in this differentiation program, but lacks DNA-binding activity until much later as the preadipocytes traverse the G(1)-S checkpoint of mitotic clonal expansion. Dominant-negative CHOP-10 is initially expressed by growth-arrested preadipocytes and sequesters/inactivates C/EBPbeta by heterodimerization with its leucine zipper. After a lag period, CHOP-10 undergoes down-regulation releasing C/EBPbeta from inhibitory constraint allowing transactivation of the C/EBPalpha and PPARgamma genes, transcription factors required for terminal differentiation. We verify that following induction of differentiation in FBS-containing medium, CHOP 10 undergoes down-regulation and differentiation occurs normally. However, when differentiation is induced in calf serum-containing medium, CHOP-10 is not down regulated resulting in delayed and incomplete differentiation. Under these conditions the expression of C/EBPalpha and PPARgamma and the accumulation of cytoplasmic triglyceride are attenuated. It appears that a factor(s) present in FBS is required to affect the down-regulation of CHOP-10 necessary for successful terminal differentiation. PMID- 16256947 TI - Mitochondrial ribosomal protein L41 mediates serum starvation-induced cell-cycle arrest through an increase of p21(WAF1/CIP1). AB - Ribosomal proteins not only act as components of the translation apparatus but also regulate cell proliferation and apoptosis. A previous study reported that MRPL41 plays an important role in p53-dependent apoptosis. It also showed that MRPL41 arrests the cell cycle by stabilizing p27(Kip1) in the absence of p53. This study found that MRPL41 mediates the p21(WAF1/CIP1)-mediated G1 arrest in response to serum starvation. The cells were released from serum starvation induced G1 arrest via the siRNA-mediated blocking of MRPL41 expression. Overall, these results suggest that MRPL41 arrests the cell cycle by increasing the p21(WAF1/CIP1) and p27(Kip1) levels under the growth inhibitory conditions. PMID- 16256948 TI - Prostaglandin E2 induces cyclooxygenase-2 expression in human non-pigmented ciliary epithelial cells through activation of p38 and p42/44 mitogen-activated protein kinases. AB - Prostaglandins (PGs) have been implicated in lowering intraocular pressure (IOP). A possible role of cyclooxygenase-2 (COX-2) in this process was emphasized by findings showing impaired COX-2 expression in the non-pigmented ciliary epithelium (NPE) of patients with primary open-angle glaucoma. The present study investigates the effect of the major COX-2 product, PGE(2), on the expression of its synthesizing enzyme in human NPE cells (ODM-2). PGE(2) led to an increase of COX-2 mRNA and protein expression, whereas the expression of COX-1 remained unchanged. Upregulation of COX-2 expression by PGE(2) was accompanied by time dependent phosphorylations of p38 mitogen-activated protein kinase (MAPK) and p42/44 MAPK, and was abrogated by inhibitors of both pathways. Moreover, PGE(2) induced COX-2 expression was suppressed by the intracellular calcium chelator, BAPTA/AM, and the protein kinase C inhibitor bisindolylmaleimide II, whereas the protein kinase A inhibitor H-89 was inactive in this respect. Induction of COX-2 expression was also elicited by butaprost (EP(2) receptor agonist) and 11-deoxy PGE(1) (EP(2)/EP(4) receptor agonist), but not by EP(1)/EP(3) receptor agonists (17-phenyl-omega-trinor PGE(2), sulprostone). Consistent with these findings, the EP(1)/EP(2) receptor antagonist, AH-6809, and the selective EP(4) receptor antagonist, ONO-AE3-208, significantly reduced PGE(2)-induced COX-2 expression. Collectively, our results demonstrate that PGE(2) at physiologically relevant concentrations induces COX-2 expression in human NPE cells via activation of EP(2)- and EP(4) receptors and phosphorylation of p38 and p42/44 MAPKs. Positive feedback regulation of COX-2 may contribute to the production of outflow facilitating PGs and consequently to regulation of IOP. PMID- 16256949 TI - Evidence in oyster of a plasma extracellular superoxide dismutase which binds LPS. AB - We have characterized in the oyster Crassostrea gigas an extracellular superoxide dismutase (Cg-EcSOD) which appears to bind lipopolysaccharides (LPS). The protein has been purified from the oyster plasma and identified as a Cu/ZnSOD according to its N-terminal sequencing and biological activity. Cg-EcSOD expression and synthesis are restricted to hemocytes as revealed by in situ hybridization and immunocytochemistry. Cg-EcSOD-expressing hemocytes were seen in blood circulation, in connective tissues, and closely associated to endothelium blood vessels. Cg-EcSOD presents in its amino acid sequence a LPS-binding motif found in the endotoxin receptor CD14 and we show that the protein displays an affinity to Escherichia coli bacteria and with LPS and Lipid A. Additionally, an RGD motif known to be implicated in the association to membrane integrin receptor is present in the amino acid sequence. The purified Cg-EcSOD was shown to bind to oyster hemocytes and to be immunocolocalized with a beta-integrin-like receptor. PMID- 16256950 TI - Development of a single-cassette system for spatiotemporal gene regulation in mice. AB - The tetracycline-regulated gene expression system has been widely used in mice to turn a transgene on and off in a target organ, but with only limited success. We developed an advanced system in which a Tet-off regulation unit was integrated into the ROSA26 locus and became active after Cre-mediated excision of the neo(r) gene. We examined the utility of this system through regulable expression of the homeodomain transcription factor pdx-1 and enhanced green fluorescent protein. The resulting mice showed strict tetracycline-regulable gene expression in all the organs where the neo(r) gene had been removed. When combined with organ specific Cre recombinase transgenic mice, our system allows us to manipulate the gene expression in an organ-specific and temporal manner. This Tet-off system should serve as an efficient tool to analyze the roles of genes in complex biological systems, such as embryogenesis, metabolism, immune system, etc. PMID- 16256951 TI - The serine protease Sp7 is expressed in blood cells and regulates the melanization reaction in Drosophila. AB - Serine proteases play a central role in defense against pathogens by regulating processes such as blood clotting, melanization of injured surfaces, and proteolytic activation of signaling pathways involved in innate immunity. Here, we present the functional characterization of the Drosophila serine protease Sp7 (CG3006) by inducible RNA interference. We show that Sp7 is constitutively expressed in blood cells during embryonic and larval stages. Silencing of the gene impairs the melanization reaction upon injury. Our data demonstrate that Sp7 is required for phenoloxidase activation and its activity is restricted to a subclass of blood cells, the crystal cells. Transcriptional up-regulation of Sp7 was observed after clean, septic injury and in flies expressing an activated form of Toll; however, mutations in the Toll or the IMD pathway did not abolish expression of Sp7, indicating the existence of other regulatory pathways and/or independent basal transcription. PMID- 16256952 TI - Theoretical studies of Alzheimer's disease drug candidate 3-[(2,4 dimethoxy)benzylidene]-anabaseine (GTS-21) and its derivatives. AB - Theoretical and molecular modeling studies have been conducted for understanding the details of how 3-[(2,4-dimethoxy)benzylidene]-anabaseine dihydrochloride (GTS 21) and its metabolism derivatives bind with the receptor of alpha7 nicotinic acetylcholine dimer. Good accordance with experimental results has been achieved. It was found that the van der Waals repulsion makes the dominant contribution to the binding energy. GTS-21 and its metabolites are apparently too large for the binding sites of the alpha7 dimer. To improve the effectiveness of the drug, a possible approach is to reduce its volume while maintaining the presence of the active groups. Our studies, in combination with experimental studies, will lead to a promising basis for practical drug design against Alzheimer's disease. PMID- 16256953 TI - Identification of SepCRP analogues in the cuttlefish Sepia officinalis: a novel family of ovarian regulatory peptides. AB - In the cuttlefish, Sepia officinalis, the ovary appears to be one of the main sources of regulatory peptides involved in the successive steps of egg-laying. Following the identification of the SepCRP-1, which is a peptide extracted from ovary and involved in egg capsule secretion, investigations were focused on the identification of related peptides. Seven related-Sepia Capsule Releasing Peptides (R-SepCRPs) were identified by means of mass spectrometry and characterized using MS/MS spectra and/or Edman degradation. Finally, primary structures were verified by the comparison of MS/MS spectra from endogenic and synthetic peptides. This new ovarian peptide family exhibits a conserved SLXKD tag involved in the biological activity. LC-MS/MS screening clearly demonstrates that R-SepCRPs are restricted to the female genital tract. Expressed during vitellogenesis, they are released by vitellogenic follicles and full-grown oocytes (FGO) in the genital coelom. Biological activities suggest that R-SepCRPs would be responsible for the storage of FGO before mating and would take part in the mechanical secretion of egg capsule products, as previously described for SepCRP-1. PMID- 16256954 TI - Prediction of protein signal sequences and their cleavage sites by statistical rulers. AB - Functioning as an "address tag" or "zip code" that guides nascent proteins (newly synthesized proteins in the cytosol) to wherever they are needed, signal peptides (also called targeting signals or signal sequences) have become a crucial tool in finding new drugs or reprogramming cells for gene therapy. To effectively and timely use such a tool, however, the first important thing is to develop an automated method for quickly and accurately identifying the signal peptide for a given nascent protein. With the avalanche of new protein sequences generated in the post-genomic era, the challenge has become even more urgent and critical. In this paper, five statistical rulers were derived via performing a mutual information analysis. By combining these statistical rulers, a new prediction algorithm was established and high success prediction rates were observed. The new algorithm may play a complementary role to the existing algorithms in this area. It is anticipated that the mutual information approach introduced here may be very useful for studying many other sequence-coupling problems in molecular biology as well. PMID- 16256955 TI - Inhibition of lipopolysaccharide-induced cyclooxygenase-2 transcription by 6 (methylsulfinyl) hexyl isothiocyanate, a chemopreventive compound from Wasabia japonica (Miq.) Matsumura, in mouse macrophages. AB - 6-(Methylsulfinyl)hexyl isothiocyanate (6-MITC) is a chemopreventive compound occurring in Wasabi (Wasabia japonica (Miq.) Matsumura), which is a very popular pungent spice in Japan. We investigated the effects of 6-MITC on the expression of cyclooxygenase-2 (COX-2) in lipopolysaccharide (LPS)-activated murine macrophage RAW264 cells. Treatment with 6-MITC suppressed LPS-mediated induction of COX-2 protein in a dose-dependent manner. Transfections with various COX-2 promoter reporter constructs revealed that the inhibitory effects of 6-MITC on COX-2 gene expression were directed by the core promoter elements including nuclear factor kappaB (NF-kappaB), CCAAT/enhancer-binding protein (C/EBP) and cyclic AMP-response element (CRE) sites. Western blotting analysis showed that 6 MITC inhibited LPS-induced activation of MAPK (ERK, p38 kinase and JNK) and transcriptional factors (CREB, c-Jun and C/EBPdelta) binding the core elements of COX-2 promoter, substantiating the involvement of these signal transduction pathways in the regulation of COX-2 expression by 6-MITC. Moreover, Western blotting experiments with MAPK-specific inhibitors (U0126 for MEK1/2, SB203580 for p38 kinase and SP600125 for JNK) demonstrated that 6-MITC suppressed LPS induced COX-2 expression by blocking the activation of JNK-mediated AP-1 and ERK/p38 kinase-mediated CREB or C/EBPdelta. Finally, the structure-activity study revealed that the inhibitory potency of methylsulfinyl isothiocyanates (MITCs) depended on the methyl chain length. These findings demonstrate for the first time that 6-MITC is an effective agent to attenuate COX-2 production, and enhance our understanding of the anti-inflammation properties of 6-MITC. PMID- 16256956 TI - Association between amygdala hyperactivity to harsh faces and severity of social anxiety in generalized social phobia. AB - BACKGROUND: Previous functional brain imaging studies of social anxiety have implicated amygdala hyperactivity in response to social threat, though its relationship to quantitative measures of clinical symptomatology remains unknown. The primary aim of this study was to examine the association between response to emotionally harsh faces in the amygdala, a region implicated in social and threat related processing, and severity of social anxiety symptoms in patients with generalized social phobia (GSP). METHODS: Ten subjects with GSP naive to psychotropic medications and without psychiatric comorbidity and ten healthy comparison subjects matched on age, gender, ethnicity, and education completed the Liebowitz Social Anxiety Scale and underwent high-field (4Tesla) functional magnetic resonance imaging while viewing blocks of emotionally salient faces. RESULTS: Relative to happy faces, activation of the amygdala in response to harsh (angry, disgusted, fearful) faces was greater in GSP patients than in controls, and the extent of amygdala activation was positively correlated with severity of social anxiety symptoms, but not general state or trait anxiety levels. CONCLUSIONS: Our findings suggest that amygdala activation to interpersonal threat can be specifically linked to the severity of social anxiety symptoms of individual GSP patients, and thus, may serve as a useful functional marker of disease severity. PMID- 16256957 TI - Immune activation during pregnancy in mice leads to dopaminergic hyperfunction and cognitive impairment in the offspring: a neurodevelopmental animal model of schizophrenia. AB - BACKGROUND: Maternal viral infection is associated with increased risk for schizophrenia. It is hypothesized that the maternal immune response to viruses may influence fetal brain development and lead to schizophrenia. METHODS: To mimic a viral infection, the synthetic double strand RNA polyriboinosinic polyribocytidilic acid (poly I:C) was administered into pregnant mice. Behavioral evaluations (thigmotaxis, methamphetamine [MAP]-induced hyperactivity, novel object recognition test [NORT]), sensorimotor gating (prepulse inhibition [PPI]), and biochemical evaluation of the dopaminergic function of the offspring of phosphate-buffered saline (PBS)-treated dams (PBS-mice) and that of poly I:C treated dams (poly I:C-mice) were examined. RESULTS: In juveniles, no difference was found between the poly I:C-mice and PBS-mice. However, in adults, the poly I:C-mice exhibited attenuated thigmotaxis, greater response in MAP-induced (2 mg/kg) hyperlocomotion, deficits in PPI, and cognitive impairment in NORT compared with the PBS-mice. Cognitive impairment in the adult poly I:C-mice could be improved by subchronic administration of clozapine (5.0 mg/kg) but not haloperidol (.1 mg/kg). Increased dopamine (DA) turnover and decreased receptor binding of D2-like receptors, but not D1-like receptors, in the striatum were found in adult poly I:C-mice. CONCLUSIONS: Prenatal poly I:C administration causes maturation-dependent increased subcortical DA function and cognitive impairment in the offspring, indicating a neurodevelopmental animal model of schizophrenia. PMID- 16256958 TI - The intrahippocampal administration of the neurosteroid allopregnanolone blocks the audiogenic seizures induced by nicotine. AB - Allopregnanolone (AlloP), GABA(A) positive modulator, has efficacy as anticonvulsant. In contrast, nicotine and pregnenolone sulfate (PregS) act as potent convulsants. The present study aims to evaluate whether a promnesic dose of PregS and/or an anxiolytic dose of AlloP administered in the hippocampus can affect the audiogenic seizures induced by nicotine administration. Rats were assigned at random to six groups that received two consecutive intrahippocampal (dorsal CA1) injections once a week during three consecutive weeks. First injection: nicotine (4.6 microg, 20 mM) or saline, second injection: PregS (5 ng, 24 microM), AlloP (0.2 microg, 1.26 microM) or saline. After the last injections, locomotor activity and audiogenic seizures were tested. AlloP decreased the horizontal and vertical activity, suggesting sedative effects. Nicotine induced behavioral convulsions and AlloP acted as an anticonvulsant. AlloP reversed the seizures induced by nicotine and decreased the audiogenic convulsions in comparison with the controls. PregS also reversed the nicotine-induced audiogenic seizures in the nicotine group but not in the control group. These results suggest that anticonvulsive effects of AlloP and PregS are mediated by different action mechanisms such as GABA(A) positive modulation, or negative modulatory action on neural nicotinic acetylcholine receptors. Even though several brain structures could be involved, these results highlight the important role played by hippocampal cholinergic and GABAergic activities, as well as neurosteroids, especially AlloP, in preventing convulsive behavior. PMID- 16256959 TI - CB1 receptor knockout mice show similar behavioral modifications to wild-type mice when enkephalin catabolism is inhibited. AB - Behavioral and biochemical studies have suggested a functional link between the endogenous cannabinoid and opioid systems. Different hypotheses have been proposed to explain the interactions between opioid and cannabinoid systems such as a common pathway stimulating the dopaminergic system, a facilitation of signal transduction- and/or a cannabinoid-induced enhancement of opioid peptide release. However, at this time, all the studies have been performed with exogenous agonists (delta-9-tetrahydrocannabinol or morphine), leading to a generally excessive stimulation of receptors normally stimulated by endogenous effectors (anandamide or opioid peptides) in various brain structures. To overcome this problem, we have measured various behavioral responses induced by the stimulation of the endogenous opioid system using the dual inhibitor of enkephalin-degrading enzymes, RB101, in CB1 receptor knockout mice. Thus, analgesia, locomotor activity, anxiety and antidepressant-like effects were measured after RB101 administration (80 and 120 mg/kg i.p. or 10 mg/kg, i.v.) in CB1 receptor knockout mice and their wild-type littermates. In all the experiments, inhibition of enkephalin catabolism produced similar modifications in behavior observed in CB1 knockout and wild-type mice. These results suggest limited physiological interaction between cannabinoid and opioid systems. PMID- 16256960 TI - Anandamide suppression of Na+ currents in rat dorsal root ganglion neurons. AB - Anandamide, the ethanolamide of arachidonic acid, is an endogenous cannabinoid. It is an agonist at CB1 and CB2 cannabinoid receptors as well as the vanilloid receptor, VR1. It is analgesic in inflammatory and neuropathic pain. Both central and peripheral mechanisms are considered to participate in its analgesia. Primary sensory neurons express Na+ currents that are involved in the pathogenesis of pain. We examined the effect of anandamide on tetrodotoxin-sensitive (TTX-S) and tetrodotoxin-resistant (TTX-R) Na+ currents in rat dorsal root ganglion neurons. Anandamide inhibited both Na+ currents in a concentration-dependent manner. At a membrane potential of -80 mV, the current inhibition was greater in TTX-S than TTX-R currents (K(d); 5.4 microM vs. 38.4 microM). The activation and inactivation became faster in TTX-R current but not in TTX-S current. Anandamide did not alter the activation voltage in either type of current. It, however, produced a hyperpolarizing shift of the steady-state inactivation voltage in both types of currents. The maximum availability at a large negative potential was not reduced by anandamide. Thus, anandamide seems to affect inactivated Na+ channels rather than resting channels. The inhibition of Na+ currents was not reversed by AM 251 (a CB1 antagonist), AM 630 (a CB2 antagonist) or capsazepine (a VR1 antagonist), suggestive of a direct action of anandamide on Na+ channels. The inhibition of Na+ currents in sensory neurons may contribute to the anandamide analgesia. PMID- 16256961 TI - Regional difference in corticotropin-releasing factor immunoreactivity in mossy fiber terminals innervating calretinin-immunoreactive unipolar brush cells in vestibulocerebellum of rolling mouse Nagoya. AB - Unipolar brush cells (UBCs), a class of interneurons in the vestibulocerebellum, play roles in amplifying excitatory inputs from vestibulocerebellar mossy fibers. This study aimed to clarify whether corticotropin-releasing factor (CRF)-positive mossy fiber innervation of calretinin (CR)-positive UBCs was altered in rolling mouse Nagoya (RMN). The distribution and the number of CR-positive UBCs in the vestibulocerebellum were not different between RMN and control mice. Double immunofluorescence revealed that some CRF-positive mossy fiber terminals were in close apposition to CR-positive UBCs. In the lobule X of vermis, such mossy fiber terminals were about 5-fold greater in number in RMN than in controls. In contrast, the number of CRF-positive mossy fiber terminals adjoining CR-positive UBCs in the flocculus was not significantly different between RMN and controls. The results suggest increased number of CRF-positive mossy fiber terminals innervating CR-positive UBCs in the lobule X but not in the flocculus of RMN. CRF may alter CR-positive UBC-mediated excitatory pathways in the lobule X of RMN and may disturb functions of the lobule X such as cerebellar adaptation for linear motion of the head. PMID- 16256962 TI - Activation of c-Jun NH2-terminal kinase 3 is mediated by the GluR6.PSD-95.MLK3 signaling module following cerebral ischemia in rat hippocampus. AB - Kainate receptor glutamate receptor 6 (GluR6) binds to the postsynaptic density protein 95 (PSD-95), which in turn anchors mixed lineage kinase 3 (MLK3) via SH3 domain in rat brain tissue. MLK3 subsequently activates c-Jun NH(2)-terminal kinase (JNK) via MAP kinase kinases (MKKs). We investigated the association of PSD-95 with GluR6 and MLK3, MLK3 autophosphorylation, the interaction of MLK3 with JNK3, and JNK3 phosphorylation following cerebral ischemia in rat hippocampus. Our results indicate that the GluR6.PSD-95.MLK3 complex peaked at 6 h of reperfusion. Furthermore, MLK3 autophosphorylation and the interaction of MLK3 with JNK3 occurred with the alteration of GluR6.PSD-95.MLK3 signaling module. To further prove whether JNK3 activation in ischemic hippocampus is mediated by GluR6.PSD-95.MLK3 signaling pathway, the AMPA/KA receptor antagonist 6,7-dinitroquinoxaline-2, (1H, 4H)-dione (DNQX), the GluR6 antagonist 6,7,8,9 Tetrahydro-5-nitro-1H-benz[g]indole-2,3-dione-3-oxime (NS102), the AMPA receptor antagonist 1-(4-aminophenyl)-4-methyl-7,8-methylenedioxy-5H-2,3-benzo diazepine (GYKI52466), and the NMDA receptor antagonist ketamine were given to the rats 20 min prior to ischemia. Our findings indicate that both DNQX and NS102 significantly attenuated the association of PSD-95 with GluR6 and MLK3, MLK3 autophosphorylation, interaction of MLK3 with JNK3, and JNK3 phosphorylation, while GYKI52466 and ketamine had no effect. Moreover, administration of NS102 before cerebral ischemia significantly increased the number of the surviving hippocampal CA1 pyramidal cells at 5 days of reperfusion. Consequently, GluR6, one subunit of kainate receptor, plays a critical role in inducing JNK3 activation after ischemic injury. PMID- 16256963 TI - Folate, vitamin E, and acetyl-L-carnitine provide synergistic protection against oxidative stress resulting from exposure of human neuroblastoma cells to amyloid beta. AB - Oxidative stress is an early and pivotal factor in Alzheimer's disease (AD). The neurotoxic peptide amyloid-beta (Abeta) contributes to oxidative damage in AD by inducing lipid peroxidation, which in turn generates additional downstream cytosolic free radicals and reactive oxygen species (ROS), leading to mitochondrial and cytoskeletal compromise, depletion of ATP, and ultimate apoptosis. Timely application of antioxidants can prevent all downstream consequences of Abeta exposure in culture, but in situ efficacy is limited, due in part to prior damage as well as difficulty in delivery. Herein, we demonstrate that administration of a combination of vitamin E (which prevents de novo membrane oxidative damage), folate (which maintains levels of the endogenous antioxidant glutathione), and acetyl-L-carnitine (which prevents Abeta-induced mitochondrial damage and ATP depletion) provides superior protection to that derived from each agent alone. These findings support a combinatorial approach in Alzheimer's therapy. PMID- 16256964 TI - Predominant fusion of bone marrow-derived cardiomyocytes. AB - OBJECTIVES: Here we address the capacity of bone marrow-derived cells (BMDCs) to trans-differentiate into mature myocytes under the physiological stimulus of exercise training. METHODS: For this purpose, we have transplanted bone marrow from mice ubiquitously expressing enhanced green fluorescence protein (eGFP) into host mice that have been subjected to a prolonged program of exercise. RESULTS: In all successful bone marrow reconstitutions (greater than 80%), we observed rare but consistent events of bone marrow-derived cardiomyocytes, the frequency of which was unchanged upon exercise training. We have further determined whether these recruited myocytes are a product of trans-differentiation or fusion by the use of a genetic system that distinguishes cell fusion from trans-differentiation in a single-cell assay. CONCLUSIONS: We concluded that both in the unchallenged mouse and in the trained specimens, fusion is the most prominent mechanism by which bone marrow-derived cells are observed in the myocyte compartment. PMID- 16256965 TI - IL-2 absorption affects IFN-gamma and IL-5, but not IL-4 producing memory T cells in double color cytokine ELISPOT assays. AB - Cytokine assays are gaining increasing importance for human immune monitoring because they reliably detect antigen-specific T cells in primary PBMC, even at low clonal sizes. Double color ELISPOT assays permit the simultaneous visualization of cells producing two different cytokines. Permitting the simultaneous assessment of type 1 and 2 immunity and due to the limited numbers of PBMC available from human study subjects, double color assays should be particularly attractive for clinical trials. Since the performance of double color assays has not yet been validated, we set out to compare them to single color measurements. Testing the recall antigen-induced cytokine response of PBMC, we found that double color assays regularly provided lower numbers of IFN-gamma and IL-5 spots than single color measurements when IL-2 detection was part of the double color assay. We showed that the inhibitory effect resulted from IL-2 absorption and could be overcome by either antibody free preactivation cultures or by inclusion of anti-CD28 antibody. In contrast, the simultaneous detection of IL-2 did not affect the numbers of IL-4 spots. Therefore, unlike IL-2/IL-4 and IFN-gamma/IL-5 assays, IL-2/IFN-gamma, and IL-2/IL-5 assays require compensation for the IL-2 capture to provide accurate numbers for the frequencies of cytokine producing memory T cells. PMID- 16256966 TI - Interactions between the peripheral site and the acylation site in acetylcholinesterase. AB - Acetylcholinesterase (AChE) hydrolyzes its physiological substrate acetylcholine at one of the highest known catalytic rates. Two sites of ligand interaction have been identified: an acylation site or A-site at the base of the active site gorge, and a peripheral site or P-site at its mouth. Despite a wealth of information about the AChE structure and the role of specific residues in catalysis, an understanding of the catalytic mechanism and the role of the P-site has lagged far behind. In recent years we have clarified how the P- and A-sites interact to promote catalysis. Our studies have revealed that the P-site mediates substrate trapping and that ligand binding to the P-site can result in steric blockade of the A-site as well as allosteric activation. We have demonstrated this activation only for the acylation step of the catalytic reaction, but others have proposed that it involves the deacylation step. To investigate this point, we have measured the reaction of carbamoyl esters (carbamates) with AChE. With these slowly hydrolyzed substrates, the carbamoylation (acylation) and decarbamoylation (deacylation) steps can be resolved and analyzed separately. Carbamoylcholine is one of the closest structural analogs of acetylcholine, and we monitored these steps in continuous mixed assays with acetylthiocholine as a reporter substrate. At high concentrations of carbamoylcholine, decarbamoylation was inhibited but no activation of carbamoylation was observed. However, high concentrations of acetylthiocholine had no effect on the decarbamoylation rate constants. We concluded that the binding of acetylthiocholine to the P-site does not activate deacylation reactions. PMID- 16256967 TI - Oxidative stress related DNA adducts in the liver of female rats fed with sunflower-, rapeseed-, olive- or coconut oil supplemented diets. AB - Both animal and epidemiological studies support an effect of fatty acid composition in the diet on cancer development, in particular on colon cancer. We investigated the modulating effect of supplementation of the diet of female F344 rats with sunflower-, rapeseed-, olive-, or coconut oil on the formation of the promutagenic, exocyclic DNA adducts in the liver, an organ where major metabolism of fatty acids takes place. 1,N(6)-ethenodeoxyadenosine (etheno-dA), 3,N(4) ethenodeoxycytidine (etheno-dC) and 1,N(2)-propandodeoxyguanosine from 4-hydroxy 2-nonenal (HNE-dGp) were determined as markers for DNA-damage derived from lipid peroxidation products and markers for oxidative stress. 8-Oxo-deoxyguanosine (8 Oxo-dG) was also measured as direct oxidative stress marker. The body weight of the rats was not influenced by the four diets containing the different vegetable oils during the 4-week feeding period. Highest adduct levels of etheno-dC (430 +/ 181 adducts/10(9) parent bases), HNE-dGp (617 +/- 96 adducts/10(9) parent bases) and 8-Oxo-dG (37,400 +/- 12,200 adducts/10(9) parent bases) were seen in rats on sunflower oil diet (highest linoleic acid content). Highest adducts levels of etheno-dA (133 +/- 113 adducts/10(9) parent bases) were found in coconut oil diet (lowest content of linoleic acid). Weakly positive correlations between linoleic acid content in the four diet groups were only observed for levels of HNE-dGp and 8-Oxo-dG. Neither the diet based on olive oil (which contains mainly oleic acid) nor the diet based on rapeseed oil (containing alpha-linolenic acid) exerted any significant protective effect against oxidative DNA damage. Our results indicate that a high linoleic acid diet may contribute to oxidative stress in the liver of female rats leading to a marginal increase in oxidative DNA-damage. PMID- 16256968 TI - Functional requirements for the optimal catalytic configuration of the AChE active center. AB - Functional analysis of the HuAChE active center architecture revealed that accommodation of structurally diverse substrates and other ligands is achieved through interactions with specific subsites such as the acyl pocket, cation binding site, hydrophobic site or the oxyanion hole. Recent studies have begun to unravel the role of this active center architecture in maintaining the optimal catalytic facility of the enzyme through inducing proper alignment of the catalytic triad. The exact positioning of the catalytic glutamate (Glu334) seems to be determined by a hydrogen bond network including several polar residues and water molecules. Disruption of this network by replacement of Ser229 by alanine is thought to remove the Glu334 carboxylate from the vicinity of His447 abolishing catalytic activity. The proper orientation of the catalytic histidine side chain is maintained by these polar interactions as well as through "aromatic trapping" by residues lining the HuAChE active center gorge. Thus, replacement of aromatic residues in the vicinity of His447, as in the F295A/F338A or in the Y72N/Y124Q/W286A/F295L/F297V/Y337A (hexamutant which mimicks the aromatic lining of HuBChE) enzymes, resulted in a dramatic decrease in catalytic activity, which was proposed to originate from catalytically nonproductive mobility of His447. Yet, HuBChE is catalytically efficient indicating that "aromatic trapping" is not the only way to conformationally stabilize the His447 side chain. A possible restriction of this mobility in a series of F295X/F338A HuAChEs was examined in silico followed by site-directed mutagenesis. Both simulations and reactivities of the actual F295X/F338A enzymes, carrying various aliphatic residues at position 295, indicate that of the bulky amino acids, like leucine or isoleucine, only methionine was capable of maintaining the catalytically viable conformation of His447. The F295M/F338A HuAChE was only two-fold less reactive than the F338A enzyme toward acetylthiocholine, and exhibited wild type-like reactivity toward covalent modifiers of the catalytic Ser203. The findings are consistent with the notion that different combinations of steric interference and specific polar interactions serve to maintain the position of His447 and thereby the high efficiency of the catalytic machinery. The two seemingly conflicting demands on the architecture of the active center-flexible accommodation of substrate and optimal juxtaposition of residues of the catalytic triad, demonstrate the truly amazing molecular design of the AChE active center. PMID- 16256970 TI - Expression of cholinesterases in brain and non-brain tumours. AB - Although the involvement of cholinesterases (ChEs) in the removal of acetylcholine (ACh) at cholinergic synapses is firmly established, there is evidence to suggest that acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) take part in several cellular processes. The early expression of ChE genes during embryonic development and their role in morphogenesis and apoptosis have been explained on the basis of the non-cholinergic actions of ChEs. In addition, the effects of AChE and BuChE, their inhibitors and antisense oligonucleotides in proliferating cellular systems, together with the mitogenic actions of ACh, support a role for ChEs in cell cycle control. The anomalous expression of ChEs may increase cell proliferation and contribute to cancer growth or development. The aim of this report is to compile the available information on ChEs in cancerous tissues in order to stimulating the research to clarify the molecular mechanisms by which ChEs may participate in cancer. Future investigations may throw light into this intriguing issue which will be of benefit to humankind. PMID- 16256969 TI - Hysteresis of butyrylcholinesterase in the approach to steady-state kinetics. AB - Butyrylcholinesterase (BChE) displays hysteretic behavior with certain neutral and charged substrates in the approach to steady state. Previous studies led us to interpret this phenomenon in terms of slow transitions between two enzyme conformers E and E'. This kinetic peculiarity is observed in human, horse and rat BChE. Oscillations that superimpose on the hysteretic lag are observed when benzoylcholine and N-alkyl derivatives of benzoylcholine are used as substrate. Hysteresis of BChE can be modulated by medium parameters (pH, salts, temperature, and pressure). Though mutant enzymes show different hysteretic behavior, so far attempts to provide a molecular mechanism of BChE hysteresis from mutagenesis studies have been unproductive. However, the substrate dependence of the hysteretic induction times, using wild-type BChE and several mutants, allowed us to build a general, mechanistic model for the hysteresis. In this model, substrate can bind to E, E', or both conformers, and ES and/or E'S can be catalytically active. The exact pathway followed depends on both the nature of the substrate and the structure of the BChE mutant under study. We propose that oscillations develop when substrate exists in different, slowly interconvertible, conformational and/or aggregation forms, of which only the minor form is capable of reacting with BChE. In support of this proposal, NMR studies have provided direct evidence for slow equilibria between monomeric and micellar forms of long chain, alkyl derivatives of benzoyl-(N-substituted) choline. There is no direct evidence that hysteresis plays a role in BChE function(s). However, the "new view" of protein dynamics proposes that proteins are normally in equilibrium between pre-existing, functional and non-functional conformers; and that binding a ligand to the functional form shifts that equilibrium towards the functional conformation. Therefore, a physiological or toxicological relevance for the hysteresis in BChE cannot be ruled out. PMID- 16256971 TI - Transcriptional regulation of acetylcholinesterase-associated collagen ColQ in fast- and slow-twitch muscle fibers. AB - The presence of a collagenous protein (ColQ) characterizes the collagen-tailed forms of acetylcholinesterase (AChE) and butyrylcholinesterase at vertebrate neuromuscular junctions, which is tethered in the synaptic basal lamina. ColQ subunits, differing mostly by their signal sequences, are encoded by transcripts ColQ-1 and ColQ-1a, which are differentially expressed in slow- and fast-twitch muscles in mammals, respectively. Both ColQ transcripts are derived from a single COLQ gene. Transcripts encoding ColQ increased during myogenic differentiation of C2C12 cells; the increase was in parallel with AChE catalytic subunit. Quantitative PCR analysis indicated that the increase during the myotube formation was due to the up regulation of ColQ-1 transcript instead of ColQ-1a. In order to reveal the regulatory mechanism of ColQ transcripts, two distinct promoters, pColQ-1 and pColQ-1a, were isolated from human COLQ gene. The ColQ promoters showed a muscle fiber type-specific expression pattern, and which was in line with the expression of endogenous transcript. After in vivo DNA transfection, pColQ-1 showed strong activity in slow-twitch muscle (e.g. soleus), while pColQ-1a was preferably expressed in fast-twitch muscle (e.g. tibialis). Mutation analysis of the ColQ promoters suggested that the muscle fiber type specific expression pattern of ColQ transcripts was regulated by a slow upsteam regulatory element (SURE) and a fast intronic regulatory element (FIRE). These results explain the specific expression patterns of collagen-tailed AChE in slow and fast muscle fibers. PMID- 16256972 TI - Effect of carbamate esters on neurite outgrowth in differentiating human SK-N-SH neuroblastoma cells. AB - Carbamate esters are widely used as pesticides and can cause neurotoxicity in humans and animals; the exact mechanism is still unclear. In the present investigation, the effects of carbamates at sublethal concentration on neurite outgrowth and cytoskeleton as well as activities of acetylcholinesterase (AChE) and neuropathy target esterase (NTE) in differentiating human SK-N-SH neuroblastoma cells were studied. The results showed that 50 microM of either aldicarb or carbaryl significantly decreased neurite length in the retinoic acid induced differentiation of the neuroblastoma cells, compared to cells treated with vehicle. Western blot analyses revealed that neither carbamate had significant effects on the levels of actin, or total neurofilament high molecular proteins (NF-H). However, increased NF-H phosphorylation was observed following carbamate treatment. These changes may represent a useful in vitro marker of carbamate neurotoxicity within a simple model of neuronal cell differentiation. Furthermore, activity of AChE, but not NTE, was significantly inhibited by aldicarb and carbaryl in differentiating cells, which suggested that cytoskeletal protein changes induced by carbamate esters in differentiating cells was associated with inhibition of AChE but not NTE. PMID- 16256973 TI - Dehydroepiandrosterone sulfate (DHEA-S) distribution in Spanish prepuberal children: relationship with fasting plasma insulin concentrations and insulin resistance. AB - BACKGROUND: The aim of this study was to analyze dehydroepiandrosterone sulfate (DHEA-S) levels in a population-based sample of Spanish prepuberal children and to investigate the relationship between DHEA-S and insulin. METHODS: 854 (440 boys and 414 girls) randomly selected prepuberal children were included in our study after a sampling. Children were 6 to 8 years old and were classified for the analysis in half-year intervals. DHEA-S and insulin levels were measured. RESULTS: DHEA-S levels increase significantly with age during prepuberty reaching the maximum level of DHEA-S for this period at 7.5 years old in girls and 8 years old in boys. Girls have significantly higher log DHEA-S levels than boys, except at the age of 8, where the levels are similar (median: 41.7 nmol/l girls and 41.1 nmol/l boys). DHEA-S correlates positively and significantly with weight, height, and BMI in all age intervals but the correlation between DHEA-S and insulin and HOMA is present only at the age of 6.5 in boys and 8 in girls. CONCLUSIONS: We report data about the distribution of DHEA-S in the Spanish prepuberal population. The maximum level of DHEA-S in this prepuberal period was reached before in girls than in boys, with girls having higher DHEA-S levels than boys until the end of this period. We found an important association between DHEA-S levels and weight, height and BMI but an inconsistent association of DHEA-S with insulin and HOMA. PMID- 16256975 TI - The functional form of the lognormal distribution as a sum of decaying exponential and sinusoidal terms applied to the isometric pinch force of human fingers. AB - The transient data of the pinch force produced between the human forefinger and thumb have been shown to fit the functional form of the well-known lognormal density function. Isometeric force generation is achieved by the stochastic recruitment of individual motor units, which sum together. Evidence from animal and human experiments demonstrates that the force generation can be modelled by underdamped terms. It is shown that a lognormal time series (distribution) can be fitted to a sum of exponential decaying sinusoidal terms. PMID- 16256974 TI - Is deoxypyridinoline a good resorption marker to detect osteopenia in phenylketonuria? AB - OBJECTIVES: To evaluate deoxypyridinoline as a resorption marker in phenylketonuria (PKU) and to search for a relationship between deoxypyridinoline, calcium/creatinine index (Ca/Cr I), osteocalcin and bone alkaline phosphatase (BAP). METHODS: This was a transversal analytical study of 46 PKU patients [17.5 (4-38) years]. Deoxypyridinoline and osteocalcin were measured with a chemiluminescent assay and BAP was measured with an immunoradiometric assay. RESULTS: Deoxypyridinoline was significantly increased in patients aged 7-14 and >18 years old, being associated with age (r=-0.724, P<0.001). Adult patients showed significantly higher Ca/Cr I, which correlates with Phe values for the year prior to the study (P=0.014). Serum BAP was significantly increased in pediatric patients (9-13 years), while it was decreased in adult patients (P=0.003). Decreased osteocalcin levels were found in patients>15 years (P=0.028). Altered deoxypyridinoline and BAP values were related (P=0.042). CONCLUSION: PKU patients excreted increased D-Pyr, suggesting high bone resorption. Bone formation seems active in childhood but deteriorates in adult PKU patients. Periodic measurement of D-Pyr and BAP may be useful in the prevention of osteopenia in PKU patients. PMID- 16256976 TI - Conditional deletion of beta-catenin in the mesenchyme of the developing mouse uterus results in a switch to adipogenesis in the myometrium. AB - Precise cell fate decisions during differentiation of uterine tissues from the embryonic Mullerian duct are critical for normal fertility. Wnt-7a, a member of the Wnt family of secreted signaling molecules that can signal through a canonical beta-catenin pathway, is necessary for the correct differentiation of both anterior/posterior and radial axes of the uterus. In order to investigate the role of beta-catenin directly in mouse uterine development, we have generated mice that are deficient in beta-catenin expression in the embryonic Mullerian duct. We have found that conditional deletion of beta-catenin in the Mullerian duct mesenchyme before postnatal differentiation of the uterine layers results in a phenotype that is distinct from the phenotype observed by deletion of Wnt-7a. Shortly after birth, the uteri of the conditional mutants appear smaller and less organized. The uteri of adult conditional beta-catenin mutants are grossly deficient in smooth muscle of the myometrium, which has been replaced by adipose, a phenotype resembling human lipoleiomyoma. We also show that the adipocytes in the uteri of mice conditionally deleted for beta-catenin are derived from Mullerian inhibiting substance type II receptor-expressing cells suggesting that they share a common origin with the uterine smooth muscle cells. These results describe the first molecular evidence linking disruption of beta-catenin expression in mesenchymal cells with a switch from myogenesis to adipogenesis in vivo. PMID- 16256977 TI - Large effects from small exposures. II. The importance of positive controls in low-dose research on bisphenol A. AB - Over six-billion pounds per year of the monomer bisphenol A (BPA) are used to manufacture polycarbonate plastic products, resins lining cans, dental sealants, and polyvinyl chloride plastic products. There are 109 published studies as of July 2005 that report significant effects of low doses of BPA in experimental animals, with many adverse effects occurring at blood levels in animals within and below average blood levels in humans; 40 studies report effects below the current reference dose of 50 microg/kg/day that is still assumed to be safe by the US-FDA and US-EPA in complete disregard of the published findings. The extensive list of significant findings from government-funded studies is compared to the 11 published studies that were funded by the chemical industry, 100% of which conclude that BPA causes no significant effects. We discuss the importance of appropriate controls in toxicological research and that positive controls are required to determine whether conclusions from experiments that report no significant effects are valid or false. PMID- 16256978 TI - Protection by a transdermal patch containing physostigmine and procyclidine of soman poisoning in dogs. AB - The prophylactic efficacy of a combinational patch system containing physostigmine and procyclidine against soman intoxication was evaluated using dogs. Female beagle dogs (body weights 9-10 kg) were shaved on the abdominal side, attached with a matrix-type patch (7x7 cm) containing 1.5% of physostigmine plus 6% procyclidine for 2 days, and challenged with subcutaneous injection of serial doses (2-10 LD50) of soman. Separately, in combination with the patch attachment, atropine (2 mg/dog) plus 2-pralidoxime (600 mg/dog) or atropine plus 1-[([4-(aminocarbonyl)pyridinio]methoxy)methyl]-2 [(hydroxyimino)methyl]pyridinium (HI-6, 500 mg/dog) were injected intramuscularly 1 min after soman poisoning. The LD50 value of soman was determined to be 9.1 microg/kg, and high doses (> or = 1.4 LD50) of soman induced salivation, emesis, defecation and diarrhea, tremors and seizures, and recumbency of dogs, leading to 100% mortality in 24 h. The prophylactic patch, which led to mean 18.5-18.8% inhibition of blood cholinesterase activity by physostigmine and mean 7.9-8.3 ng/ml of blood concentration of procyclidine, exerted a high protection ratio (4.7 LD50), in comparison with relatively-low effects of traditional antidotes, atropine plus 2-pralidoxime (2.5 LD50) and atropine plus HI-6 (2.7 LD50). Noteworthy, a synergistic increase in the protection ratio was achieved by the combination of the patch with atropine plus HI-6 (9 LD50), but not with atropine plus 2-pralidoxime (5 LD50). In addition, the patch system markedly attenuated the cholinergic signs and seizures induced by soman, especially when combined with atropine plus HI-6, leading to elimination of brain injuries and physical incapacitation up to 6 LD50 of soman poisoning. Taken together, it is suggested that the patch system containing physostigmine and procyclidine, especially in combination with atropine and HI-6, could be a choice for the quality survival from nerve-agent poisoning. PMID- 16256979 TI - CRTH2-specific binding characteristics of [3H]ramatroban and its effects on PGD2 , 15-deoxy-Delta12, 14-PGJ2- and indomethacin-induced agonist responses. AB - We previously showed that ramatroban (Baynastrade mark), a thromboxane A(2) (TxA(2)) antagonist, had inhibited prostaglandin D(2) (PGD(2))-stimulated human eosinophil migration mediated through activation of chemoattractant receptor homologous molecule expressed on Th2 cells (CRTH2). However, detailed pharmacological characterization of its inhibitory activity has not been described. In the present study, we showed that [(3)H]ramatroban bound to a single receptor site on CRTH2 transfectants with a similar K(d) value (7.2 nM) to a TxA(2) receptor (8.7 nM). We also demonstrated that ramatroban inhibited PGD(2) , 15-deoxy-Delta(12, 14)-PGJ(2) (15d-PGJ(2))- and indomethacin-induced calcium responses on CRTH2 transfectants in a competitive manner with similar pA(2) values (8.5, 8.5, and 8.6, respectively). This is the first report showing the evidence for direct binding of ramatroban to CRTH2, revealing its competitive inhibitory effects and another interesting finding that PGD(2), indomethacin and 15d-PGJ(2) share the same binding site with ramatroban on CRTH2. PMID- 16256980 TI - Acute treatment with morphine augments the expression of serine racemase and D amino acid oxidase mRNAs in rat brain. AB - To obtain further insight into the interactions between the N-methyl-D-aspartate receptor and opioid receptor systems, we have investigated the effects of the acute treatment of morphine on the expression of serine racemase and D-amino acid oxidase mRNAs in several brain areas of rats. The morphine administration produced a dose-dependent and transient elevation in the levels of serine racemase and D-amino acid oxidase mRNAs in all the brain areas. The present results are the first to suggest an interaction between the expression of the mRNAs for the D-serine-related enzymes and the opioid receptor activation. PMID- 16256981 TI - Nitric oxide contributes to AT2 but not AT1 angiotensin II receptor-mediated vasodilatation of porcine pial arteries and arterioles. AB - Angiotensin II elicits pial artery dilation by activating angiotensin AT1 and angiotensin AT2 receptors. This study determined if vasodilatation in response to angiotensin AT2 receptor activation is due to stimulated release of nitric oxide (NO) in newborn pigs equipped with a closed cranial window. Angiotensin II (10( 8), 10(-6) M) elicited pial artery dilatation that was unchanged by the NO synthase inhibitor N omega-Nitro-L-Arginine (L-NNA) (10(-6) M) (12+/-3 and 18+/-2 versus 12+/-3 and 21+/-4%). Angiotensin II was not associated with changes in artificial cerebrospinal fluid (CSF) cGMP concentration, an indicator of NO release. Similar data were obtained for the angiotensin AT1 receptor agonist L 162,313. In contrast, the angiotensin AT2 receptor agonist CGP 42112A (10(-8), 10(-6) M) induced vasodilatation that was blocked by L-NNA (9+/-2 and 18+/-3 versus 1+/-1 and 1+/-1%). CGP 42112A dilatation was associated with elevated artificial CSF cGMP concentration (757+/-18, 1590+/-89, and 2101+/-116 fmol/ml) and such stimulated release was blocked by L-NNA. These data indicate that stimulated NO release contributes to angiotensin AT2 but not angiotensin AT1 induced vasodilatation. These data suggest that angiotensin II primarily elicits dilatation via angiotensin AT1 receptor activation. PMID- 16256982 TI - Mouse organic anion transporter 2 and 3 (mOAT2/3[Slc22a7/8]) mediates the renal transport of bumetanide. AB - Multispecific organic anion transporters play an important role in the excretion and the elimination of a wide variety of endogenous and exogenous substrates. To date, five murine OAT homologs such as mouse organic anion transporters 1-3, 5, and 6 (mOAT1-3, 5 and 6) have been isolated and well characterized. With the exception of mOAT6, other mOAT isoforms are predominantly expressed in the kidney. The aim of this study was to examine whether mOAT2/3, as well as hOAT2/3, transports the diuretic bumetanide using a Xenopus laevis oocyte expression system. When expressed in Xenopus oocytes, mOAT2/3 mediated the high affinity transport of bumetanide. The apparent K(m) values for the uptake of bumetanide via mOAT2 and mOAT3 were 9.12 +/- 2.42 microM and 1.01 +/- 0.27 microM, respectively. Immunohistochemical analysis revealed that mOAT2 is expressed on the luminal membrane site of the proximal tubule. Our results indicate that mOAT2 and 3, as well as human homologs, are molecules for the transport of bumetanide on the luminal membranes of kidney proximal tubules. PMID- 16256983 TI - Regulation of mesenchymal stem cell and chondrocyte differentiation by MIA. AB - Melanoma inhibitory activity (MIA), also referred to as cartilage-derived retinoic acid-sensitive protein (CD-RAP), an 11-kDa secreted protein, is mainly expressed in cartilaginous tissue during embryogenesis and adulthood. Currently, the function of MIA in cartilage tissue is not understood. Here, we describe that MIA acts as a chemotactic factor on the mesenchymal stem cell line C3H10T1/2, stimulating cell migration significantly at concentrations from 0.24 to 240 ng/ml, while inhibiting cell migration at higher doses of 2.4 microg/ml. When analyzing the role of MIA during differentiation processes, we show that MIA by itself is not capable to induce the differentiation of murine or human mesenchymal stem cells. However, MIA influences the action of bone morphogenetic protein (BMP)-2 and transforming growth factor (TGF)-beta 3 during mesenchymal stem cell differentiation, supporting the chondrogenic phenotype while inhibiting osteogenic differentiation. Quantitative RT-PCR analysis revealed the up regulation of the cartilage markers MIA, collagen type II and aggrecan in human mesenchymal stem cell (HMSC) cultures differentiated in the presence of MIA and TGF-beta 3 or BMP-2 when compared to HMSC cultures differentiated in the presence of TGF-beta 3 or BMP-2 alone. Further, MIA down-regulates gene expression of osteopontin and osteocalcin in BMP-2 treated HMSC cultures inhibiting the osteogenic potential of BMP-2. In the case of human primary chondrocytes MIA stimulates extracellular matrix deposition, increasing the glycosaminoglycan content. Therefore, we postulate that MIA is an important regulator during chondrogenic differentiation and maintenance of cartilage. PMID- 16256984 TI - Endothelial cadherins and tumor angiogenesis. AB - Adherens junctions and more specifically cadherins play an important role in endothelial cell integrity and growth and, in general, in vascular morphogenesis. Besides their adhesive properties, cadherins may act by transferring intracellular signals through interaction with a complex network of cytoskeletal and signaling molecules. Cadherins may signal in different ways: through direct activation of signaling pathways, through interaction with cell-specific growth factor receptors or by controlling beta-catenin and/or other transcription factors' translocation to the nucleus. Endothelial cells present different cadherins which may transfer specific signals and exert distinct functional roles. VE-cadherin is endothelial-specific and the major constituent of adherens junctions. This protein is able to protect endothelial cells from apoptosis and contributes to contact inhibition of endothelial cell growth. N-cadherin is also abundantly expressed in the endothelium and may be important in modulating VE cadherin expression. T cadherin, R-cadherin and VE-cadherin 2 were found in specific regions of the vascular tree but their role in vascular development or angiogenesis is still unclear. PMID- 16256986 TI - Study of lens autofluorescence by fluorophotometry in pregnancy. AB - Lens autofluorescence originates from an accumulation of fluorescent substances such as the tryptophan-derived residues and protein aggregations, which are associated with the preclinical progress of cataractogenesis, diabetes and lens aging. Our purpose is to determine if pregnancy alters the typical constituents of the lens autofluorescence. Fifteen healthy pregnant women (22 eyes) who were in their third trimester of pregnancy and 23 age-matched healthy controls (37 eyes, non-pregnant females). Lens autofluorescence, lens transmission and corneal autofluorescence were studied with fluorophotometry. The lens autofluorescence values were 358+/-151 ng ml(-1) in the control group and 201+/-110 ng ml(-1) in the pregnants women. The difference was significant (p=0.0074). Lens transmission values were 0.93+/-0.02 ng ml(-1) in the control group and 0.94+/-0.02 ng ml(-1) in the pregnants women: the difference was not significant. Corneal autofluorescence values were 21.9+/-7.5 ng ml(-1) in the control group and 18.2+/ 5.8 ng ml(-1) in the pregnant women. The difference was not significant. Our study showed a significant decrease in lens autofluorescence in pregnant women compared to a normal population. The decrease can be partly attributed to the aqueous component of the lens that increases significantly during the final trimester of pregnancy and that this provokes a dilution of the fluorescent substances. PMID- 16256985 TI - Nested collagen matrices: a new model to study migration of human fibroblast populations in three dimensions. AB - Fibroblast-3D collagen matrix culture provides a model system to analyze cell physiology under conditions that more closely resemble tissue than conventional 2D cell culture. Previous work has focused primarily on remodeling and contraction of collagen matrices by fibroblasts, and there has been little research on migration of cell populations within the matrix. Here, we introduce a nested collagen matrix model to analyze migration of fibroblasts in 3D collagen matrices. Nested collagen matrices were prepared by embedding contracted cell containing matrices (also called dermal equivalents) inside cell-free matrices; migration occurred from the former to the latter. Control experiments with human dermal fragments in place of dermal equivalents confirmed the reliability of the model. Human fibroblast migration in nested collagen matrices occurred after a lag phase of 8-16 h, and cells migrating out of the inner matrices were bipolar with leading dendritic extensions. Migration was myosin II, Rho kinase and metalloproteinase-dependent but did not require plasma fibronectin. Platelet derived growth factor but not lysophosphatidic acid or serum stimulated cell migration, although all three of these physiological agonists promote matrix remodeling and contraction. The nested collagen matrix model is a relatively easy, rapid and quantitative method to measure migration of cell populations. Our studies using this model demonstrate important differences between regulation of fibroblast migration and remodeling in collagen matrices. PMID- 16256987 TI - Axial myopia induced by a monocularly-deprived facemask in guinea pigs: A non invasive and effective model. AB - This study evaluated the efficacy of a facemask, a non-invasive and potentially more reliable method, in inducing axial myopia in guinea pigs. Thirty-six animals were randomly assigned to 3 groups: MDF (monocularly-deprived facemask, n=6), lid suture (eyelids sutured monocularly, n=24) and normal control (free of form deprivation, n=6). All the groups underwent biometric measurement (refraction, corneal curvature and axial length) prior to the experiment. All animals in the MDF group underwent biometric measurement at each of the 4 timepoints (2, 4, 6 and 8 weeks of form deprivation). In the lid-sutured group, the animals were randomly assigned to 4 subgroups (n=6 each) and each subgroup underwent biometric measurement at one of the timepoints matching those of the MDF group. In the normal control group, all animals underwent biometric measurement at each of the timepoints matching those of the 2 experimental groups. Placement of a facemask on an animal took approximately 10 sec and all the facemasks remained in place at all timepoints. The procedure of lid-suture took at least 20 min for an animal and rupture of the sutures occurred in 50% of the animals after 4 weeks. The MDF eyes developed myopia from -2.21+/-2.11D (Mean+/-s.d.) at 2 weeks to -4.38+/-2.14 at 8 weeks (p<0.05 at all timepoints, compared to the contralateral eyes) with a lengthening of the vitreous chamber from 0.17+/-0.05 mm at 2 weeks to 0.29+/-0.12 mm at 8 weeks (p<0.01 at all timepoints, compared to the contralateral eyes). The lid-sutured eyes developed myopia from -2.38+/-1.21D at 2 weeks to -4.75+/-1.39D at 8 weeks (p<0.05 at all timepoints, compared to the contralateral eyes) with a lengthening of the vitreous chamber from 0.13+/-0.02 mm at 2 weeks to 0.30+/-0.10 mm at 8 weeks (p<0.05 at 2, 4, 8 weeks, but >0.05 at 6 weeks, compared to the contralateral eyes) and an increase in the radius of the corneal curvature (0.20+/-0.07 mm at 4 weeks, p<0.01; 0.17+/-0.05 mm at 8 weeks, p<0.05; compared to the contralateral eyes). Both the MDF and lid-sutured groups had a similar development in myopia and vitreous length (MDF vs lid-suturing: p>0.05 at all timepoints, one-way ANOVA with Bonferroni correction). This development was significantly faster than in the normal control group (MDF or lid-suture vs normal control: p<0.05 to <0.01 from 2 to 8 weeks, one-way ANOVA with Bonferroni correction). The radius of corneal curvature in the lid-sutured group was significantly greater than in either the MDF group or the normal control group since 4 weeks of form deprivation (p<0.05, one-way ANOVA with Bonferroni correction). Treatment with MDFs is as effective as the lid-suture in inducing axial myopia in guinea pigs. This method is non-invasive and allows evaluation of the same group of animals at different timepoints so that the number of animals required could be minimized without affecting the accuracy of the results. PMID- 16256988 TI - Plasmodium chabaudi: reverse transcription PCR for the detection and quantification of transmission stage malaria parasites. AB - We have developed two reverse transcription polymerase chain reaction (RT-PCR) techniques to detect and quantify the transmission stages (gametocytes) of Plasmodium chabaudi malaria parasites. Both the qualitative and quantitative techniques are based on the amplification of mRNA coding for the P. chabaudi protein Pcs230, which is expressed exclusively in gametocytes. The quantitative RT-PCR (qRT-PCR) technique was developed and validated by examining serial dilutions of known gametocyte densities. The method generated a high correlation between calibration curves of blind samples (R(2)=0.86). The technique was found to be specific, reproducible, and time efficient for quantification of both patent and sub-patent gametocytemia with a sensitivity level 100-1000 times greater than microscopy. The qualitative RT-PCR (RT-PCR) technique was used to monitor the persistence and dynamics of P. chabaudi gametocytes following acute infection. Mice in two independent experiments were sampled for up to 87 days post-infection. RT-PCR showed that gametocytes can persist for up to 8 weeks, post-infection, whereas microscopy could only detect gametocytes up to 6 weeks. Potential applications of the above techniques for studying the ecology, evolution, and epidemiology of malaria transmission are discussed. PMID- 16256989 TI - Gonadotropin-releasing hormone in invertebrates: structure, function, and evolution. AB - Gonadotropin-releasing hormone (GnRH) is central to the initiation and maintenance of reproduction in vertebrates. GnRH is found in all major groups of Phylum Chordata, including the protochordates. Studies on functional and structural evolution of GnRH have, in the past, focused exclusively on chordates. However, the recent structural elucidation of an octopus GnRH-like molecule and increasing evidence that GnRH-like substances are present in multiple invertebrate phyla suggest GnRH is an ancient peptide that arose prior to the divergence of protostomes and deuterostomes. The extraordinary conservation of GnRH structure and function raises interesting questions regarding the functional role assumed by GnRH over the course of evolution. This review will focus on the current understanding of GnRH structure and function in non-chordate invertebrates. Special emphasis will be placed upon the possible and speculated functions of GnRH in mollusks. PMID- 16256990 TI - Elevated corticosterone levels elicit non-calling mating tactics in male toads independently of changes in circulating androgens. AB - Competition among males for a limited number of females may result in the expression of condition-dependent alternative mating tactics. In such cases, decision rules mediating mating tactic expression are likely to be influenced by a male's external as well as internal environment. For example, experimental studies with anuran amphibians (frogs and toads) indicate that changes in the social-acoustic environment alter the probability that an individual adopts a calling versus non-calling "satellite" mating tactic. However, there is considerable variation in the behavioral responses of individuals in such studies, suggesting that physiological differences among individuals play an important role in tactic expression. For instance, recent models predict that natural elevation in adrenal glucocorticoids during vocal production alters androgen production and/or neural activity to mediate transitions between reproductive tactic expression in anuran amphibians. Using corticosteroid injections, we show that elevation in circulating corticosterone levels significantly increase the probability that free-ranging male Great Plains toads (Bufo cognatus) and Woodhouse's toads (B. woodhousii) adopt a non-calling satellite tactic. Corticosterone-induced behavioral transitions occurred rapidly (<1 h) and independently of fluctuations in circulating androgen levels, suggesting a direct effect of glucocorticoids on brain centers controlling vocal production. We discuss our findings in the context of behavioral studies that alter the social-acoustic environment to examine its influence on tactic expression. PMID- 16256991 TI - Experimentally increased social competition compromises humoral immune responses in house finches. AB - Although social behavior can substantially influence an individual's physiology, few studies have examined whether intraspecific competition compromises individual immunocompetence. We experimentally manipulated the intensity of social competition in captive non-breeding house finches (Carpodacus mexicanus) by supplying few (high competition) or many (low competition) feeding sites. We tested whether elevated levels of social competition caused individual changes in aggression rates, humoral immunity, body mass, and baseline and stress-induced corticosterone concentrations. We also examined whether physiological responses to social competition were related to an individual's social status. We found that house finches under high social competition had significantly higher aggression rates, lower antibody responses, and lost more body mass. Within flocks, dominant individuals mounted stronger immune responses in both competition treatments. Our statistical power to detect differences in circulating corticosterone concentrations was low, but we did not find any support for the hypothesis that corticosterone concentrations mediate immunosuppression among or within flocks: baseline and stress-induced corticosterone concentrations did not differ under high and low social competition, were unrelated to individual social status, and did not predict the extent of immunosuppression among individuals. Overall, we documented that two universal components of social behavior, intraspecific competition and social status, modulated the strength of a humoral immune response in house finches. PMID- 16256992 TI - Differential effect of kinase A and C blockers on lordosis facilitation by progesterone and its metabolites in ovariectomized estrogen-primed rats. AB - Dose response curves for lordosis behavior was obtained for progesterone (P) and its two ring A-reduced metabolites: 5alpha-pregnanedione (alpha-DHP) and 5alpha,3alpha-pregnanolone (5alpha,3alpha-Pgl) by infusing these progestins in the right lateral ventricle (rlv) of ovariectomized (ovx) estradiol-treated rats (2 microg estradiol benzoate; EB), 40 h before intracerebro-ventricular (icv) injection. Effective doses 50 (ED50) revealed that ring A-reduced progestins were more potent than P itself to induce lordosis behavior. Two dose levels, one producing the maximal effect and the other one producing a submaximal response (ED50-ED60), were selected for testing the capacity of RpAMPS, a kinase A blocker, and H7, a kinase C blocker, to modify the response to the three progestins. rlv injection of RpAMPS significantly depressed the lordosis response to the two dose levels of P and alpha-DHP but failed to significantly inhibit that of 5alpha,3alpha-Pgl. The administration of H7 prevented the effect of both 5alpha-reduced progestins without affecting the response to P. The results suggest that P and its ring A-reduced metabolites stimulate lordosis behavior through different cellular mechanisms: P acting mainly through the cAMP-kinase system; alpha-DHP through both kinase A and kinase C signaling pathways and 5alpha,3alpha-Pgl through the kinase C system. PMID- 16256993 TI - Do Kirschner wires maintain reduction of displaced Colles' fractures? AB - Fifty-three patients underwent closed reduction and longitudinal k-wiring of displaced Colles' fractures and were reviewed after a mean of 26 months. Radiographs taken at the time of injury, after reduction and k-wiring, and at fracture union were compared for radial shortening and dorsal angulation. Manipulation significantly improved fracture position (P<0.001). Dorsal angulation was successfully corrected by manipulation in 98%, and this position was maintained to fracture union in all cases. Seventy-three percentage of fractures manipulated for radial shortening >2 mm were adequately reduced, but 41% of these fractures subsequently lost position to malunite. In this group of patients, the mean radial shortening between reduction and fracture union was 1.6 mm. This did not correlate with Frykman Class or radial shortening at injury. Closed reduction and k-wire stabilisation is an attractive technique because it is relatively non-invasive compared with plating or external fixation. However, a degree of radial shortening between reduction and fracture union must be anticipated. Fractures reduced inadequately to allow for this loss of radial length, are more likely to malunite. This may compromise functional outcome. PMID- 16256994 TI - Correlation between radiological parameters and patient-rated wrist dysfunction following fractures of the distal radius. AB - The present study investigates the correlation between radiological parameters of wrist fractures and the clinical outcome expressed by objective clinical parameters and the level of patient-rated wrist dysfunction. Thirty consecutive cases of unstable distal radial fractures treated with closed reduction and percutaneous fixation were prospectively studied for a period of one year. The outcome parameters included objective clinical and radiological parameters and the previously described and validated patient-rated wrist evaluation (PRWE) score. Analysis showed that for unstable (AO classification types 23-A2, -A3, -C1 and -C2) fractures the fracture type affects the range of wrist palmarflexion (p=0.04) and that the presence of postoperative articular 'step-off' affects the range of wrist dorsiflexion and the patient-rated wrist function at the final time of the study (p<0.01 and p=0.02, respectively). It is also shown that permanent radial shortening and loss of the palmar angle were associated with prolonged wrist pain (p<0.01 and p=0.03, respectively). Our finding that residual articular incongruity correlates with persisting loss of wrist dorsiflexion and wrist dysfunction contradicts the view that loss of articular congruity is associated with late development of articular degeneration but not with early wrist dysfunction. Additionally, this study failed to show any association between the fracture type and the functional outcome as rated by the patients. PMID- 16256995 TI - Corrective osteotomy for symptomatic scaphoid malunion. AB - A prospective study aimed at assessment of the outcome of management of symptomatic scaphoid malunion (hump-back deformity). The work included 13 scaphoid malunions in 13 patients. All patients complained of weak painful hand grip and limitation of wrist function. All cases were subjected to a corrective opening wedge scaphoid osteotomy with insertion of a trapezoid-shaped tricortical iliac bone graft. At the final assessment, after a mean follow-up period of 42 months, the achieved results were rated excellent in seven cases, good in four and fair in two according to the scoring system used. Objectively, the mean range of wrist motion and hand grip strength improved from 48% and 47% pre-operatively to 82% and 79% at the final assessment. Radiological parameters including height to length ratio, lateral intrascaphoid angle and dorsal cortical angle were also effectively improved. The intra-operative corrected carpal alignment has almost been maintained at the final follow-up. The procedure did not have serious drawbacks such as non-union or avascular necrosis and perhaps might delay the development of degenerative arthritis of the wrist. PMID- 16256996 TI - Forearm fractures in children. Single bone fixation with elastic stable intramedullary nailing in 20 cases. AB - We present our experience with elastic stable intramedullary nailing (ESIN) used in the single bone fixation of both bones forearm fractures in children. From May 2002 to July 2004, 20 children (14 boys and 6 girls), median age of 10 years (range 6-15 years) were treated with ESIN for 16 closed and 4 grade I open forearm fractures. All patients were reviewed clinically at a median follow-up of 20 months (range 6-30 months). All fractures were radiologically united at a median of 6.7 weeks (6-9 weeks). The median operating time was 35min (range 25 60min). The median hospital stay was 2 days (range 1-3 days). Removal of the nails was undertaken in all 20 children at a median of 19 weeks (range 16-24 weeks) post-operatively. At follow-up, a full range of elbow and wrist movements were found in all cases. There was no clinically significant rotational deformity in any case. ESIN seems to be a safe method in the treatment of single bone fixation of both bones forearm fractures in children between 6 and 15 years of age. PMID- 16256997 TI - Treatment of bilateral open calcaneal fractures with ilizarov frames. PMID- 16256998 TI - Conservative therapy of stage I endometrial adenocarcinoma and atypical endometrial hyperplasia for the preservation of fertility. PMID- 16256999 TI - LDL oxidized with iron in the presence of homocysteine/cystine at acidic pH has low cytotoxicity despite high lipid peroxidation. AB - Fe(III) can have a strong oxidizing effect in the presence of reductants at acidic pH, which may occur under anaerobic conditions or in regions of inflammation. Low density lipoprotein (LDL) oxidation with Fe(III) and homocysteine/cystine at acidic pH provoked mainly formation of lipid hydroperoxides and thiobarbituric acid reactive substances (TBARS) in the absence of significant protein modification. Even when oxidized to a high TBARS content, LDL oxidized at acidic pH was not cytotoxic when added to THP-1 monocytes in a concentration causing cell death when LDL was oxidized to a similar TBARS content at plasma pH with Fe(III) or Cu(II) in the presence or absence of homocysteine/cystine. Inducible nitric oxide production by RAW264.7 mouse macrophages was only weakly inhibited by LDL oxidized at acidic pH, even if acetylated before oxidation to increase uptake, as compared to LDL oxidized with Cu(II) at plasma pH to a similar TBARS content or anodic electrophoretic mobility. LDL oxidized at acidic pH may mainly induce protective mechanisms against oxidative stress while causing little acute damage of cells. PMID- 16257000 TI - Preparation and evaluation of novel stationary phases for improved chromatographic purification of pneumocandin B0. AB - Preparation and evaluation of a number of stationary phases for improved chromatographic purification of pneumocandin B0, a key intermediate in the synthesis of the antifungal agent, Cancidas, has led to the identification of several materials with potential for improved performance. PMID- 16257001 TI - Mass spectrometry/gas chromatography-mass spectrometry approach for rapid screening/quantitative determination of perchloroethylene in air. AB - A new mass spectrometry/gas chromatography-mass spectrometry (MS/GC-MS) approach has been developed for the screening and quantitative determination of perchloroethylene (PERC) in workplace and outdoor air samples, which could be extended to the screening and analysis of other analytes and samples. This approach may be rapidly modified in order to be used directly as an MS detector for screening purposes or alternatively as a common GC-MS, for confirmation. The screening alternative by MS is approximately 20 times faster than the quantitative-confirmatory determination by GC-MS. Detection limits of both alternatives are sufficiently low to screen and determine PERC in the above mentioned matrixes. The advantage of this approach over others previously described is that, in the present case, the sample passes through the chromatographic column only when the confirmatory GC-MS is used. For the MS screening method, the chromatographic column is bypassed by using an appropriate selection valve. In this way, the column lifetime is extended and screening time is considerably shortened. PMID- 16257003 TI - Development of a multiresidue method for analyzing herbicide and fungicide residues in bovine milk based on solid-phase extraction and liquid chromatography tandem mass spectrometry. AB - This report describes a liquid chromatography (LC)-tandem mass spectrometry (MS) multiresidue method for determining traces of 30 base/neutral/acid herbicides and fungicides in bovine whole milk. Four milliliters of milk was spiked with the analytes and two surrogate analytes (SAs) and then diluted with 35 mL of a water/methanol solution (50:50, v/v). This mixture was passed through a 0.5-g Carbograph 4 cartridge. After washings, analytes were re-extracted by back flushing the cartridge with 1.5 mL of methanol followed by 6 mL of methylene chloride/methanol (80:20, v/v), 50 mmol/L formic acid. After partial solvent removal down to about 0.1, 0.15 mL of 1 mmol/L formic acid aqueous solution and an internal standard (IS) were added. After filtration, 50 microL of the final extract was then introduced into the LC analytical column. During the chromatographic run, the MS system was operated in both positive and negative ion modes. MS data acquisition was performed in the multi-reaction monitoring mode, selecting two precursor ion>product ion transitions for each target compound, except for pentachlorophenol. On analyzing six milk samples from different sources, absolute recovery of the analytes and the two SAs ranged between 78% and 104% with RSDs not larger than 13%. The accuracy of the method at three different spike levels was assessed by adding the two SAs to analyte-containing milk samples and varied between 82% and 120% with RSDs not larger than 11%. Limits of quantification were estimated to range between 0.008 and 1.4 microg/L. Compared to the Carbograph 4 cartridge, one filled with a N-vinylpyrrolidone-m divinylbenzene co-polymer (Oasis HLB) sorbent was much less efficient in recovering several of the acidic herbicides considered and, in addition, its relative final extract produced a severe negative matrix effect that drastically weakened ion signal intensities of several non acidic analytes. PMID- 16257002 TI - Simultaneous determination of carbamate and organophosphorus pesticides in fruits and vegetables by liquid chromatography-mass spectrometry. AB - A liquid chromatography-mass spectrometry (LC-MS) method was established for the purpose of simultaneous determination of carbamate and organophosphorus (OPPs) pesticides in fruits and vegetables. Samples were extracted with acetonitrile; and then prepared by dispersive solid-phase extraction (dispersive-SPE) with primary secondary amine (PSA) as the sorbent. Four common representative samples (tomato, apple, carrot, and cabbage) were selected from the supermarket to investigate the effect of different matrices on pesticides recoveries and assay precision after spiking samples with 0.05 mg/kg. Matrix composition did not interfere significantly with the determination of the pesticides. The obtained recoveries were, with a few exceptions, in the range of 70-110% with RSDs less than 8%. It was applied to pesticide residue monitoring in vegetables and fruits from local markets. PMID- 16257004 TI - Ultra-rapid separation of an angiotensin mixture in nanochannels using shear driven chromatography. AB - The present paper reports on the separation of a mixture of fluorescein isothiocyanate-labeled angiotensin I and II peptides in a shear-driven nanochannel with a C18-coating and using an eluent consisting of 5% acetonitrile in 0.02 M aqueous phosphate buffer at pH 6.5. The flat-rectangular nanochannel in fused silica consisted of an etched structure in combination with a flat moving wall. The very fast separation kinetics that can be achieved in a nanochannel allowed to separate the angiotensin peptides in less then 0.2 s in a distance of only 1.8 mm. Plate heights as small as 0.4 microm were calculated after substraction of the injection effect. PMID- 16257005 TI - Droplet migration in emulsion systems measured using MR methods. AB - The migration of emulsion droplets under shear flow remains a largely unexplored area of study, despite the existence of an extensive literature on the analogous problem of solid particle migration. A novel methodology is presented to track the shear-induced migration of emulsion droplets based on magnetic resonance imaging (MRI). The work is in three parts: first, single droplets of one Newtonian fluid are suspended in a second Newtonian fluid (water in silicone oil (PDMS)) and are tracked as they migrate within a Couette cell; second, the migration of emulsion droplets in Poiseuille flow is considered; third, water-in silicone oil emulsions are sheared in a Couette cell. The effect of (a) rotational speed of the Couette, (b) the continuous phase viscosity, and (c) the droplet phase concentration are considered. The equilibrium extent of migration and rate of migration increase with rotational speed for two different emulsion systems and increased continuous phase viscosity, leads to a greater equilibrium extent of migration. The relationship between the droplet phase concentration and migration is however complex. These results for semi-concentrated emulsion systems and wide-gap Couette cells are not well described by existing models of emulsion droplet migration. PMID- 16257006 TI - Differential extraction of eosinophil granule proteins. AB - Eosinophil granules contain several toxic cationic proteins that contribute to the pathophysiology of allergic diseases. These include eosinophil peroxidase, two ribonucleases, and two forms of the major basic protein (MBP). Extraction of eosinophil granules by exposure to acid solution and fractionation on Sephadex G 50 characteristically yields a distinctive profile of three discrete peaks, and these proteins are usually recovered in good quantities, except for the eosinophil major basic protein homolog (MBP2). We investigated the effect of multiple granule extractions by dilute HCl on the recovery of granule proteins. Isolated granules were repetitively extracted, up to 31 times, in 0.01 M HCl, and the extracts fractionated on Sephadex G-50. Whereas initial extracts yielded the characteristic three-peak fractionation pattern, later extracts yielded four discrete peaks. Characterization of the novel fourth peak showed that it contained MBP2. These results indicate that repetitive extraction of eosinophil granules yields an increased amount of all granule proteins, and that MBP2 can now be recovered in good quantities and in a relatively pure form. PMID- 16257007 TI - The two cysteine-rich head domains of minicollagen from Hydra nematocysts differ in their cystine framework and overall fold despite an identical cysteine sequence pattern. AB - Synthetic replicates of naturally occurring cysteine-rich peptides such as hormones, neurotransmitters, growth factors, enzyme inhibitors, defensins and toxins often can be oxidatively folded in high yields to their native structure in simple redox buffers. Thereby, identical cysteine patterns in the sequence were found to generate identical disulfide connectivities and homologous spatial structures despite significant variability in the non-cysteine positions. Minicollagen-1 from the nematocysts of Hydra is a trimeric protein that contains cysteine-rich domains at the N and C termini, which are involved in the assembly of an intermolecular disulfide network. Determination of the three-dimensional structures of peptides corresponding to the N-terminal and C-terminal domains by NMR spectroscopy revealed a remarkable exception from the general rule. Despite an identical cysteine pattern, the two domains of minicollagen-1 form different disulfide bridges and exhibit distinctly different folds, both of which are not found in the current structural databases. To our knowledge, this is the first case where two relatively short peptides with the abundant cysteine residues in identical sequence positions fold uniquely and with high yields into defined, but differing, structures. Therefore, the cysteine-rich domains of minicollagen constitute ideal model systems for studies of the interplay between folding and oxidation in proteins. PMID- 16257008 TI - Oligomerization and interacellular localization of the glycoprotein receptor ERGIC-53 is independent of disulfide bonds. AB - ERGIC-53 is a type I transmembrane lectin facilitating the efficient export of a subset of secretory glycoproteins from the endoplasmic reticulum. Previous results have shown that ERGIC-53 is present as reduction-sensitive homo oligomers, i.e. as a balanced mixture of disulfide-linked hexamers and dimers, with the two cysteine residues located close to the transmembrane domain playing a crucial role in oligomerization. Here, we demonstrate, using sucrose gradient sedimentation, cross-linking analyses, and non-denaturing gel electrophoresis, that ERGIC-53 is present exclusively as a hexameric complex in cells. However, the hexamers exist in two forms, one as a disulfide-linked, Triton X-100, perfluoro-octanic acid, and SDS-resistant complex, and the other as a non covalent, Triton X-100, perfluoro-octanoic acid-resistant, but SDS-sensitive, complex made up of three disulfide-linked dimers that are likely to interact through the coiled-coil domains present in the luminal part of the protein. In contrast to what was previously believed, neither of the membrane-proximal cysteine residues plays an essential role in the formation, or maintenance, of the latter form of hexamers. Subcellular fractionation revealed that the double cysteine mutant was present in the endoplasmic reticulum-Golgi-intermediate compartment, indicating that the two cysteine residues are not essential for the intracellular distribution of ERGIC-53. Based on these results, we present a model for the formation of the two hexameric forms. PMID- 16257009 TI - Neural correlates of impulsive responding in borderline personality disorder: ERP evidence for reduced action monitoring. AB - Patients with borderline personality disorder (BPD) are characterized by marked impulsive behaviour. The impulsive response style of patients with BPD may be associated with diminished action monitoring, which can be investigated by measuring the error-related negativity (ERN). The ERN is an ERP component generated in the anterior cingulate cortex (ACC) following erroneous responses. Behavioural and ERP measurements were obtained during performance on a speeded two-choice reaction task in a group of patients with BPD (N=12) and in a group of age-matched controls (N=12). The ERP results showed that ERN amplitudes were reduced for patients with BPD, as were the P300 amplitudes after late feedback. The behavioural results confirmed a more impulsive response style for the BPD group, reflected in larger RT differences between correct and incorrect responses and in an increase in erroneous responses to the easy congruent stimuli. Additionally, analyses on post-error congruency effects demonstrated that controls adjusted their behaviour following errors, but patients with BPD did not. The attenuated ERNs indicate reduced action monitoring in patients with BPD. This suggests that the ACC, or the action-monitoring network it is part of, is not functioning optimally. Due to this reduced action monitoring, patients with BPD do not learn from their errors as well as controls. Consequently, they do not adjust their behaviour when necessary and thus maintain their impulsive response style. PMID- 16257010 TI - Polarity and hydrophobicity interactions in protein synthesis process. AB - About 30 years ago, experiments found that there are polarity and hydrophobicity (P and H) correlations and affinity between amino acids and their anticodons. Although it is shown that these experimental findings are important for explaining the origins of the genetic code, the great potential of P and H interactions in investigating other bio-functions have not been fully explored. Here, through raising, discussing and answering seven relevant questions hidden in tRNA aminoacylation, the formation of peptide bonds, and the ending of translations, etc., we show our theoretical findings that the P and H correlations and affinity take vital roles in the protein synthesis process. We found the relationship between the 3' end ACCN sequences of tRNA molecules and the activated amino acids and its biological significance, the rRNAs' consensus sequences 5'NCC...TGG3' or 5'TGG...NCC3' which may perform as functional segments of rRNAs to help triggering the reaction of peptide formation, and common nature of releasing factors that the first amino acid residue of releasing factors ERF, RF1 and RF2 are all Methionine, except a few Alanine, which may be necessary for releasing the translated polypeptide and stopping the translating process. In the terms of P and H correlations and affinity, we provide explanations of why only using the poly (G) as mRNA template cannot get the poly (Gly) in experiments deciphering the genetic code, why Gly often appears in beta turns and why translational bypassing might occur when translating 5'GGAUGA on mRNA. Since amino acids and nucleotides are the subunits, respectively, for composing proteins and nucleic acids, these findings will help in further understanding interactions among the bio-macromolecules. These findings are also helpful for investigating rRNAs, further understanding the protein synthesis process and analysing similar bio-problems, and should be proved useful for experimental biologists. PMID- 16257011 TI - Oral administration of soy-derived genistin suppresses lipopolysaccharide-induced acute liver inflammation but does not induce thymic atrophy in the rat. AB - Genistein, the principal isoflavone present in soy, has been identified as a protein tyrosine kinase (PTK) inhibitor that has in vitro anti-inflammatory effects. Whether genistein has in vivo anti-inflammatory effects remains unknown yet. Injecting or feeding rats with the unconjugated form of genistein (aglycone) results in decreased thymic weight and lymphocytopenia. However, 95-99% of genistein is present as the conjugated form genistin (genistein glycoside) in soy or soy-derived products. This study was undertaken to reveal whether genistin, as well as genistein, has anti-inflammatory effects in vivo. After oral administration of equimolar genistein (namely 7.4 or 74 micromol/dose) at daily doses of 2.0 or 20 mg/kg, or genistin at daily doses of 3.2 or 32 mg/kg for 3 days to male rats, both aglycone and glycoside suppressed the production of lipopolysaccharide (LPS)-induced tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-1beta, and IL-6 in both from the liver and in the sera. Aglycone induced thymic atrophy while glycoside did not. In vitro preincubation of liver slices from naive rat with genistein aglycone or glycoside suppressed LPS-induced TNF-alpha production in a dose-dependent manner. Taken together, both in vivo and in vitro administration of genistin and genistein suppressed LPS-induced liver pro-inflammatory cytokine production. However, equimolar oral administration of genistin did not induce thymus atrophy. Further investigation in long-term isoflavone intake is required especially among neonates. The results suggest that the safety evaluation of the consumption of isoflavone should be based on isoflavone glycoside but not aglycone. PMID- 16257012 TI - Inhibition of human telomerase reverse transcriptase gene expression by gambogic acid in human hepatoma SMMC-7721 cells. AB - The activation of human telomerase, a process regulated by the human telomerase reverse transcriptase (hTERT), is a crucial step during cellular immortalization and malignant transformation. We have reported that gambogic acid (GA), a natural product isolated from the gamboge resin of Garcinia hanburyi tree, is an effective telomerase inhibitor and thus displays potent anticancer activity both in vitro and in vivo. Here we present the direct interaction of GA with oncogene c-MYC, a ubiquitous transcription factor involved in the control of cell proliferation and differentiation, as the molecular mechanism of GA's inhibitory effect on telomerase activity. Consistent with the recently reported association between c-MYC overexpression and induction of telomerase activity, we find here that GA treatment of a human hepatoma cell line SMMC-7721 significantly reduced the expression of c-MYC in a time- and concentration-dependent manner accompanied with the down-regulation of the hTERT transcription and the ultimate reduction in telomerase activity. Our results indicate that the hTERT is a target of c-MYC activity and identify a feasible mechanism of GA's potent anticancer activity. PMID- 16257013 TI - Anthropogenic influence on the organic fraction of sediments in two contrasting estuaries: a biochemical approach. AB - Abundance and biochemical composition of organic materials in sediments of the estuaries of Mundaka and Bilbao (SE Bay of Biscay) were analysed to assess the effect of organic wastes released to these systems. Organic and labile contents were higher in the sediments of Bilbao, denoting organic enrichment with poorly decomposed materials by human dumping. Spatially, organic matter distribution skewed seaward in Bilbao and upward in Mundaka, in agreement with the location of major point sources of sewage. Labile material, proteins and protein:carbohydrate ratio showed upward increases in both estuaries, attributed to the oxygen restrictive conditions to decompose organic materials. Vertical distribution of organic components into sediments reflected the history of human activities. In Bilbao, the significant reduction of organic and labile compounds, and the decrease of protein:carbohydrate values in top layers seem a result of recent remedial actions to reduce urban sewage inputs and improve environmental conditions. Higher contributions of proteins and lipids were associated with anthropogenic materials, and higher contributions of carbohydrates with autochthonous materials. High protein:carbohydrate values indicate poorly decomposed materials of human origin. The close relationship of carbohydrates with chloropigments in Mundaka suggests that phytobenthic populations and derived detritus contributed to a greater extent to the organic fraction in this estuary. PMID- 16257014 TI - Benthic impacts of fish farming: meta-analysis of community and geochemical data. AB - A number of 41 papers dealing with the benthic effects of fish farming were reviewed and the values of the variables studied were extracted to be used in a meta-analysis of effects. The papers used covered a wide range of farmed species, geographic regions, management practices and specific site characteristics (e.g., depth, exposure, and sediment type). Therefore, the total data-set may not be considered as biased towards a particular set of conditions as is often the case with data collected in a single study. More than 120 biological and geochemical variables were monitored, occasionally using different sampling and analytical protocols for the same variables. The rank correlation analysis between all possible pairs of variables in the data set showed a large number of significant positive or negative correlations, reflecting the response of these variables to benthic organic enrichment. The use of stepwise regression showed that most biological and geochemical variables are determined by a combination of distance from the farm with bottom depth and/or latitude. Results of stepwise regression, repeated separately for each type of sediment, showed that although the general pattern was similar among different types of sediments, the coefficients varied considerably indicating changes of the distance affected by settling particulate organic material for different sediment types. The overall conclusion is that the complicated interactions between variables and the lack of data, such as current speed, induce difficulties in setting common or uniform environmental quality standards for benthic effects of fish farming and these should take into account the existing considerable differences between geographic regions, depth zones and sediment types. PMID- 16257015 TI - Impact of digenean parasite infection on metallothionein synthesis by the cockle (Cerastoderma edule): a multivariate field monitoring. AB - Metallothioneins (MT) are proteins that play an important role in metabolism of essential metals and detoxification of trace metals from living organisms. Their synthesis is induced by metal pollution but can also be exacerbated by other factors such as reproduction processes. In this context, we monitored MT concentrations in a cockle Cerastoderma edule (marine bivalve) population and highlighted the effect of a castrating digenean parasite, Labratrema minimus. In spent cockles, MT levels were low (ca. 5 nmol sites g(-1), fresh weight) but slightly higher in parasitized individuals. During gametogenesis, MT synthesis increased in all cockles, but concentrations were lower in parasitized individuals (18 against 27 nmol sites g(-1), fw in unparasitized cockles) in relation with gonad damage by parasites. Therefore, it is suggested that parasite infection in cockles can modulate MT synthesis that could consequently interfere with the response of these protective proteins in case of metal contamination. PMID- 16257016 TI - Bioaccumulation of polychlorinated biphenyls in mullet fish in a former ship dismantling harbour, a contaminated estuary, and nearby coastal fish farms. AB - This study investigated the bioaccumulation of polychlorinated biphenyls (PCBs) toward mullet fish (Liza macrolepis) living in former PCB contaminated areas, the Ann-Ping harbour and the Er-Jen estuary, and fish farms located near the above two areas in 2003. The PCB body burdens of collected fish samples are proportional to the contamination level of their locations with the following rank order (greatest to least) from the Er-Jen estuary, the Ann-Ping harbour to the fish farms. Concentration of PCBs of the estuarine mullet has been approximately decreased to one-half of the peak concentration of the 1990s. Although the concentration of PCBs in farmed fish inhabiting near the two contaminated areas was greater than the average of those of fish from local fish markets in Taiwan, no particularly great contamination level was observed in their bodies. Using the less chlorinated PCB fraction (triCB + tetraCB)/total PCBs as the indicator of the origins of PCBs, fish near former contaminated areas had greater body burdens of the more chlorinated PCB congeners, while the farmed fish exhibited a PCB pattern more like that known to originate from air-water exchange with less chlorinated PCBs predominating. Although the PCB contamination has been stopped for a decade, the residual contaminants, supposedly existing in soil and sediments, still contribute to the body burden of fish residing in the estuary and the harbour. PMID- 16257017 TI - A combined modelling and geochemical study of the fate of terrigenous inputs from mixed natural and mining sources in a coral reef lagoon (New Caledonia). AB - Open-cast mining for Ni, Cr and Co was conducted in the south-west part of New Caledonia during the 20th century. Abandoned mining and prospecting sites were severely affected by erosion, resulting in an increase in the load of terrigenous particles transported to the coral reef lagoon. This article assesses the impact of a typical small catchment area (La Coulee River, 85 km2 watershed) on two bays (Boulari and Sainte Marie) located near Noumea, New Caledonia's main city. This multi-disciplinary study combines geochemical, sedimentological, and hydrodynamic approaches. Ni and Cr concentrations contained in the geochemical matrix phase of the pelitic fraction were determined. The study of the geochemical signatures together with sedimentological data and 3D numerical simulations of the transport of non-settling particles throughout the lagoon demonstrated that terrigenous inputs from the Coulee River were mainly transported and deposited along the shoreline, reaching areas as distant as Sainte Marie Bay. Although quantitatively low (about 3% of the pelite mass of the bay sediments), the terrigenous inputs in Sainte Marie Bay originating from the Coulee River could be traced. The metal content in suspended matter was over 7000 mg kg(-1) for Ni and 4200 mg kg(-1) for Cr. PMID- 16257018 TI - Oral serotonin administration affects the quantity and the quality of macronutrients selection in European sea bass Dicentrarchus labrax L. AB - Teleost fish are able to regulate their energy intake selecting from pure macronutrients sources, but the regulatory mechanisms involved in macronutrients selection remain unknown. Serotonin (5-HT) reduces food intake in mammals and fish and modifies the macronutrients selection pattern in mammals; however, no information is available about its role on macronutrients selection in fish. The aim was to determine the effect of orally administered 5-HT (0.1, 0.5 and 2.5 mg kg BW(-)(1)) into gelatine capsules on the subsequent macronutrient selection of sea bass, using for this purpose gelatine capsules including carbohydrates, protein, or lipids separately. The voluntary ingested 5-HT was released into the plasma of fish, reaching a level two times greater than the controls, 45 min after the ingestion of a capsule containing 2.5 mg kg BW(-1) of 5-HT. The indoleamine, at doses of 0.1, 0.5 and 2.5 mg kg BW(-1), produced a reduction in total food intake of 31%, 49% and 37%, respectively, compared to the baseline, modifying the macronutrient selection pattern. The percentage of fat selected was significantly reduced whereas the percentage of protein significantly increased after administration of highest dose, but no changes were observed in the proportion of carbohydrate for any 5-HT doses. In conclusion, oral administration of 5-HT affected both amount of food intake and pattern of macronutrients selected. This is the first evidence supporting a role of 5-HT as a neurohumoral mediator involved in macronutrients selection in fish. PMID- 16257019 TI - Neural bases of food-seeking: affect, arousal and reward in corticostriatolimbic circuits. AB - Recent studies suggest that there are multiple 'reward' or 'reward-like' systems that control food seeking; evidence points to two distinct learning processes and four modulatory processes that contribute to the performance of food-related instrumental actions. The learning processes subserve the acquisition of goal directed and habitual actions and involve the dorsomedial and dorsolateral striatum, respectively. Access to food can function both to reinforce habits and as a reward or goal for actions. Encoding and retrieving the value of a goal appears to be mediated by distinct processes that, contrary to the somatic marker hypothesis, do not appear to depend on a common mechanism but on emotional and more abstract evaluative processes, respectively. The anticipation of reward on the basis of environmental events exerts a further modulatory influence on food seeking that can be dissociated from that of reward itself; earning a reward and anticipating a reward appear to be distinct processes and have been doubly dissociated at the level of the nucleus accumbens. Furthermore, the excitatory influence of reward-related cues can be both quite specific, based on the identity of the reward anticipated, or more general based on its motivational significance. The influence of these two processes on instrumental actions has also been doubly dissociated at the level of the amygdala. Although the complexity of food seeking provides a hurdle for the treatment of eating disorders, the suggestion that these apparently disparate determinants are functionally integrated within larger neural systems may provide novel approaches to these problems. PMID- 16257020 TI - Biosynthesis of the xanthophyll plectaniaxanthin as a stress response in the red yeast Dioszegia (Tremellales, Heterobasidiomycetes, Fungi). AB - Carotenoid biosynthesis was examined in a phylloplane yeast identified by ITS, 18S and 28S rDNA analysis as a Dioszegia sp. close to D. takashimae. In well aerated flask or fermentor cultures, this strain produced essentially a single pigment confirmed as the xanthophyll plectaniaxanthin by NMR analysis, at concentrations of 103-175 microgg(-1) biomass dry weight. Detailed studies showed increases in plectaniaxanthin concentrations in the presence of 5 mM hydrogen peroxide (1.8-fold), 50 and 100 microM duroquinone (3.1- and 3.7-fold, respectively), and 2% ethanol (4.9-fold). Whereas oxidative stress is known to enhance the biosynthesis of torularhodin or astaxanthin in other red yeasts where they are associated with an antioxidant function, this is the first report implicating plectaniaxanthin in a similar role. At reduced aeration, biosynthesis of plectaniaxanthin was suppressed and its putative precursor gamma-carotene accumulated. The carotenoid cyclase inhibitor nicotine (5-20 mM) inhibited plectaniaxanthin formation, with lycopene accumulating stoichiometrically. Hydroxy groups at C-1' and C-2' therefore seem to be introduced late in plectaniaxanthin biosynthesis, following cyclization of the beta-ionone ring. PMID- 16257021 TI - Comparison of different anthelminthic drug regimens against Mansonella perstans filariasis. AB - Mansonella perstans filariasis is widely present in Africa and equatorial America and its pathogenicity has recently been reconsidered. Effective treatment is lacking and there is no consensus on the optimal therapeutic approach. The aim of this study was to compare the effects of different drug regimens on M. perstans infection. Six different anthelminthic therapeutic protocols were undertaken on 165 subjects with M. perstans infection and their effects on microfilariae burden were evaluated. Diethylcarbamazine (DEC) was able to reduce microfilariae density in the majority of cases, but it seldom eliminated infection after a single treatment. Mebendazole appeared to be more active than DEC in eliminating the infection, with a comparable rate of overall responses. Ivermectin and praziquantel showed no modification of microfilariae concentration. Thiabendazole showed a small but significant activity against the infection. Combination treatments (DEC plus mebendazole) resulted in a significantly higher activity than the single drugs. PMID- 16257022 TI - Mammomonogamus roundworm (Nematoda: Syngamidae) recovered from the duodenum of a Thai patient: a first and unusual case originating in Thailand. AB - A pair of Mammomonogamus laryngeus roundworms in copula was recovered from the duodenum of a 72-year-old male Thai patient from Kanchanaburi Province. Eggs were also found in the stool of the patient. This is the first case of Mammomonogamus infection originating in Thailand, as the previous two reports from Thailand attributed the infection as originating in Malaysia. The occurrence of adult worms in the duodenum is unusual and differs from previous findings in the larynx, posterior pharynx, tracheal wall and bronchi. This adds a new dimension to the manifestation of the infection. PMID- 16257023 TI - Integrating HIV management for children into the Integrated Management of Childhood Illness guidelines. AB - The WHO/UNICEF strategy of Integrated Management of Childhood Illness (IMCI) has been adopted as a strategy for improving paediatric care in resource-poor settings. The original IMCI guidelines recommend referral of children with severe or recurrent illnesses such as those common in HIV/AIDS, however the specific identification and management was not included. In many countries, especially in sub-Saharan Africa, HIV/AIDS contributes significantly to child morbidity and mortality. There was a need to include the specific assessment and management of symptomatic HIV infection in IMCI guidelines. A draft HIV component of the IMCI guidelines was developed, which included management of symptomatic HIV cases with referral for counselling and testing, and was evaluated in South Africa. A revised version was then validated in Ethiopia and Uganda where HIV, malnutrition and malaria prevalence is different from South Africa. IMCI materials have been adapted to include an HIV component. Currently, very few children under 15 years of age receive antiretroviral therapy (ART). Chronic HIV care of children, including ART for children, has been developed within the Integrated Management of Adolescent and Adult Illness (IMAI) initiative of the WHO. Through this initiative, the WHO is assisting countries to facilitate care and treatment of children undergoing ART. PMID- 16257024 TI - Mixed infection with Leishmania (Viannia) braziliensis and Leishmania (Leishmania) chagasi in a naturally infected dog from Rio de Janeiro, Brazil. AB - We report here the first case of co-infection with Leishmania (Viannia) braziliensis and Leishmania (Leishmania) chagasi in a naturally infected dog from Rio de Janeiro, Brazil. Isoenzyme characterisation identified the parasites isolated in culture from the cutaneous lesion as L. (V.) braziliensis and the isolates from blood and lymph node as L. (L.) chagasi. PCR analysis using specific primers followed by molecular hybridisation for direct Leishmania species identification in tissue fragments confirmed the presence of L. (V.) braziliensis DNA in the cutaneous lesion and of L. (L.) chagasi DNA in spleen and popliteal lymph node fragments. This report emphasises the importance of identification of Leishmania species infecting seropositive dogs in endemic areas, and the consequent re-assessment of control and epidemiological surveillance measures for the control of leishmaniasis, as is the case in Brazil. PMID- 16257026 TI - Increased density but not prevalence of gametocytes following drug treatment of Plasmodium falciparum. AB - We monitored post-treatment Plasmodium falciparum among patients treated with chloroquine (CQ) and sulfadoxine-pyrimethamine (SP; Fansidar in a village in eastern Sudan. Parasites were examined on day 0 (pre-treatment), day 7, day 14 and day 21 (post-treatment) during the transmission season. A further sample was taken 2 months later (day 80) at the start of the dry season. Asexual forms and gametocytes were detected by microscopy, and reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect expression of gametocyte-specific proteins pfs 25 and pfg 377. Gametocyte carriage, as revealed by microscopy, increased significantly following CQ and SP treatment, reaching a maximum between days 7 and 14. When measured by RT-PCR, however, there was no significant difference in gametocyte rate between day 0 and days 7 or 14. RT-PCR gametocyte rates dropped dramatically by day 80 post treatment but were still 33% and 8% in the CQ- and SP-treated group at this time. Alleles associated with drug resistance of P. falciparum to chloroquine (the chloroquine resistance transporter, pfcrt, and multidrug resistance, pfmdr1) and to pyrimethamine (dihydrofolate reductase, dhfr) were seen at a high frequency at the beginning of treatment and increased further through time following both drug treatments. Infections with drug-resistant parasites tended to have higher gametocyte prevalence than drug-sensitive infections. PMID- 16257027 TI - Nutrient-limited food webs with up to three trophic levels: feasibility, stability, assembly rules, and effects of nutrient enrichment. AB - Community structure is controlled, among multiple factors, by competition and predation. Using the R* rule and graphical analysis, we analyse here the feasibility, stability and assembly rules of resource-based food webs with up to three trophic levels. In particular, we show that (1) the stability of a food web with two plants and two generalist herbivores does not require that plants' resource exploitation abilities trade-off with resistance to the two herbivores, and (2) food webs with two plants and either one generalist herbivore and a carnivore or two generalist herbivores and two generalist carnivores are not feasible because of cascade competition between top consumers. The relative strength of species interactions and the relative impacts of plants and herbivores on factors which control their growth also play a critical role. We discuss how community structure constrains assembly rules and yields cascades of extinctions in food webs. PMID- 16257025 TI - Characterisation of metabolic acidosis in Kenyan children admitted to hospital for acute non-surgical conditions. AB - Metabolic acidosis is associated with most severe malaria deaths in African children, and most deaths occur before maximum antimalarial action is achieved. Thus, specific acidosis treatment may reduce mortality. However, the underlying mechanisms remain poorly understood and no specific interventions have been developed. A detailed characterisation of this acidosis is critical in treatment development. We used the traditional and Stewart's approach to characterise acidosis in consecutive paediatric admissions for malaria and other acute non surgical conditions to Kilifi District Hospital in Kenya. The overall acidosis prevalence was 21%. Gastroenteritis had the highest prevalence (61%). Both the mean albumin-corrected anion gap and the strong ion gap were high (>13 mmol/l and >0 mmol/l, respectively) in malaria, gastroenteritis, lower respiratory tract infection and malnutrition. Presence of salicylate in plasma was not associated with acidosis but was associated with signs of severe illness (odds ratio 2.11, 95% CI 1.1-4.2). In malaria, mean (95% CI) strong ion gap was 15 (14-7) mmol/l, and lactate, creatinine and inorganic phosphorous explained only approximately 40% of the variability in base excess (adjusted R2 = 0.397). Acidosis may be more common than previously recognised amongst paediatric admissions in Africa and is characterised by the presence of currently unidentified strong anions. In malaria, lactate and ketones, but not salicylate, are associated with acidosis. However, unidentified anions may be more important. PMID- 16257028 TI - Factors influencing subcellular localization of the human papillomavirus L2 minor structural protein. AB - Two structural proteins form the capsids of papillomaviruses. The major structural protein L1 is the structural determinant of the capsids and is present in 360 copies arranged in 72 pentamers. The minor structural protein L2 is estimated to be present in twelve copies per capsid. Possible roles for L2 in interaction with cell surface receptors and in virion uptake have been suggested. As previously reported, L2 localizes in subnuclear domains identified as nuclear domain 10 (ND10). As it was demonstrated that L2 is able to recruit viral and cellular proteins to ND10, a possible role for L2 as a mediator in viral assembly has been proposed. In this study, we determined factors influencing the localization of L2 at ND10. Under conditions of moderate L2 expression level and in the absence of heterologous viral components, we observed that, in contrast to previous reports, L2 is mainly distributed homogeneously throughout the nucleus. L2, however, is recruited to ND10 at a higher expression level or in the presence of viral components derived from vaccinia virus or from Semliki Forest virus. We observed that translocation of L2 to ND10 is not a concentration-dependent accumulation but rather seems to be triggered by yet unidentified cellular factors. In contrast to HPV 11 and 16 L2, the HPV 18 L2 protein seems to require L1 for efficient nuclear accumulation. PMID- 16257030 TI - Working memory can guide pop-out search. AB - Top-down feedback from working memory (WM) can exert an early and involuntary influence on visual selection for targets that are relatively difficult to discriminate [Soto, D., Heinke, D., Humphreys, G. W., & Blanco, M. J. (2005) Journal of Experimental Psychology: Human Perception and Performance, 31, 248]. Here, we demonstrate similar effects even on search for a pop-out target. At the beginning of each trial, participants memorized a prime that could contain either the search target or a distracter in the subsequent search array. Targets and distractors were easily discriminable. Despite this, the prime in WM affected responses latencies and the direction of the first saccade. Top-down search, guided by the contents of WM, can modulate selection even when salient bottom-up cues are present. PMID- 16257029 TI - Widely varying SIV prevalence rates in naturally infected primate species from Cameroon. AB - Although it is now well established that a substantial proportion of wild-living primates in sub-Saharan Africa harbor SIV, no study to date has examined to what extent the various species are naturally infected. In this study, we first describe the development and validation of sensitive and specific SIV antibody detection assays representing all major known primate lentiviral lineages on a panel of 207 sera from 11 different primate species with known infection status. The newly developed assays were then used to determine SIV prevalence rates in nine primate species native to Cameroon. Analysis of 722 sera revealed widely varying prevalence rates, ranging from an apparent absence of SIV infection in crested mona (0/70), grey cheeked (0/36) and agile mangabeys (0/92), to prevalence rates of 3%, 4%, 11%, 27%, 39% and 52% for mustached (6/203), greater spot-nosed (8/193), northern talapoin (3/26), mantled guereza (14/52), De Brazza's (9/23) and mandrill (14/27) monkeys, respectively. The epidemiology of naturally occurring SIV infections is thus more complex than previously appreciated and the various non-human primate hosts seem to differ in their susceptibility to SIV infection. The newly developed assays should now permit to define with greater accuracy existing SIV reservoirs and associated human zoonotic risk. PMID- 16257031 TI - Depth aftereffects mediated by vertical disparities: evidence for vertical disparity driven calibration of extraretinal signals during stereopsis. AB - Perceptual adaptation often results in a repulsive aftereffect: stimuli are seen as biased away from the adaptation stimulus (). Here we report the absence of a repulsive aftereffect for a vertical gradient of vertical disparity (or vertical size ratio, VSR). We exposed observers to a binocular stimulus consisting of horizontal lines. This stimulus contains vertical, but not horizontal disparities. The visual system was able to measure the VSR of this stimulus: although the lines themselves always appeared unslanted, the VSR carried by the lines had a dramatic effect on the apparent slant of a horizontal row of dots, as predicted by recent accounts of Ogle's (1938) induced effect (e.g., Backus, Banks, van Ee, & Crowell, 1999). Yet we observed no repulsive aftereffect for the VSR signal: after adaptation to horizontal lines that were vertically larger in one eye, we found an attractive aftereffect, the magnitude of which was largest in stimuli that did not contain a VSR signal. We interpret these results as a case of recalibration: disagreement between extra-retinal eye position signals (EP) and VSR causes a recalibration in the use of EP as used in the stereoscopic perception of slant. PMID- 16257032 TI - Processing of global form and motion in migraineurs. AB - Previous studies have identified anomalies of cortical visual processing in migraineurs that appear to extend beyond V1. Migraineurs respond differently than controls to transcranial magnetic stimulation of V5, and can demonstrate impairments of global motion processing. This study was designed to assess the integrity of intermediate stages of both motion and form processing in people with migraine. We measured the ability to integrate local orientation information into a global form percept, and to integrate local motion information into a global motion percept. Control subjects performed significantly better than migraineurs on both tasks, suggesting a diffuse visual cortical processing anomaly in migraine. PMID- 16257033 TI - Transformation and characterisation of dissolved organic matter during the thermophilic aerobic biodegradation of faeces. AB - We conducted a comparison of the characteristics of dissolved organic matter (DOM) taken from the bio-toilet and other sources. A characterisation of DOM was carried out to assess the stability of the compost generated during the thermophilic and aerobic biodegradation of faeces. In addition, levels of soluble microbial products generated in the bio-toilet composting reactor were compared with those taken from other sources. The results showed that (i) the main component of DOM from the bio-toilet are solutes with molecular weight (MW)>30,000 Da (40%), whereas micromolecules (MW< 1000 Da) constituted more than 60% of the DOM from other solid samples, while liquid samples reached even more than 90%; (ii) the DOM stabilisation level in the composting reactor of the bio toilet system was greater than that shown by DOM from other sources; (iii) stabilisation of DOM in the bio-toilet system was characterised by an increasing amount of macromolecules (MW>30,000 Da) after a decreasing trend was observed in the early stages of the biodegradation process; and (iv) net production of lipopolysaccharide (LPS) in wastewater treatment plants is greater than in the bio-toilet. PMID- 16257034 TI - Decrease in the relative heterogeneity of health with age: a cross-national comparison. AB - Using data from seven studies from Canada, Australia and the United States we were able to evaluate the heterogeneity of health status for 34,095 people aged 60 years and older. We found that relative heterogeneity (measured by the coefficient of variation) decreased with age. The coefficient of variation also decreased as a function of deficit accumulation (i.e. was lower when people demonstrated more deficits) displaying a power-law relationship. The exponent was close to 0.5, and therefore belongs to the dynamic universality class, which is typical for complex dynamical networks. We interpret this as showing that decline in relative heterogeneity is a robust finding, and represents, at the group level, increased vulnerability amongst elderly people. PMID- 16257035 TI - Differential modulation of base excision repair activities during brain ontogeny: implications for repair of transcribed DNA. AB - DNA repair sustains fidelity of genomic replication in proliferating cells and integrity of transcribed sequences in postmitotic tissues. The repair process is critical in the brain, because high oxygen consumption exacerbates the risk for accumulation of oxidative DNA lesions in postmitotic neurons. Most oxidative DNA damage is repaired by the base excision repair (BER) pathway, which is initiated by specialized DNA glycosylases. Because the newly discovered Nei-like mammalian DNA glycosylases (NEIL1/2) proficiently excise oxidized bases from bubble structured DNA, it was suggested that NEILs favor repair of transcribed or replicated DNA. In addition, since NEILs generate 3'-phosphate termini, which are poor targets for AP endonuclease (APE1), it was proposed that APE1-dependent and independent BER sub-pathways exist in mammalian cells. We measured expression and activities of BER enzymes during brain ontogeny, i.e., during a physiologic transition from proliferative to postmitotic differentiated state. While a subset of BER enzymes, exhibited declining expression and excision activities, expression of NEIL1 and NEIL2 glycosylases increased during brain development. Furthermore, the capacity for excision of 5-hydroxyuracil from bubble structured DNA was retained in the mature rat brain suggesting a role for NEIL glycosylases in maintaining the integrity of transcribed DNA in postmitotic brain. PMID- 16257036 TI - Abatement of mercury pollution in the small-scale gold mining industry: restructuring the policy and research agendas. AB - This paper critiques contemporary research and policy approaches taken toward the analysis and abatement of mercury pollution in the small-scale gold mining sector. Unmonitored releases of mercury from gold amalgamation have caused considerable environmental contamination and human health complications in rural reaches of sub-Saharan Africa, Latin America and Asia. Whilst these problems have caught the attention of the scientific community over the past 15-20 years, the research that has since been undertaken has failed to identify appropriate mitigation measures, and has done little to advance understanding of why contamination persists. Moreover, the strategies used to educate operators about the impacts of acute mercury exposure, and the technologies implemented to prevent further pollution, have been marginally effective at best. The mercury pollution problem will not be resolved until governments and donor agencies commit to carrying out research aimed at improving understanding of the dynamics of small scale gold mining communities. Acquisition of this knowledge is the key to designing and implementing appropriate support and abatement measures. PMID- 16257037 TI - Are pharmaceuticals potent environmental pollutants? Part I: environmental risk assessments of selected active pharmaceutical ingredients. AB - As part of achieving national environmental goals, the Swedish Government commissioned an official report from the Swedish Medical Products Agency on environmental effects of pharmaceuticals. Considering half lives/biodegradability, environmental occurrence, and Swedish sales statistics, 27 active pharmaceutical ingredients were selected for environmental hazard and risk assessments. Although there were large data gaps for many of the compounds, nine ingredients were identified as dangerous for the aquatic environment. Only the sex hormones oestradiol and ethinyloestradiol were considered to be associated with possible aquatic environmental risks. We conclude that risk for acute toxic effects in the environment with the current use of active pharmaceutical ingredients is unlikely. Chronic environmental toxic effects, however, cannot be excluded due to lack of chronic ecotoxicity data. Measures to reduce potential environmental impact posed by pharmaceutical products must be based on knowledge on chronic ecotoxic effects of both active pharmaceutical ingredients as well as excipients. We believe that the impact pharmaceuticals have on the environment should be further studied and be given greater attention such that informed assessments of hazards as well as risks can be done. PMID- 16257039 TI - Elevated methylmercury concentrations and loadings during flooding in Minnesota rivers. AB - Previous studies have identified flooded landscapes (e.g., wetlands, impoundments) as sites of elevated methylmercury (MeHg) production. Here we report MeHg and total Hg (THg) concentrations and mass loadings in rivers in Minnesota during major flooding episodes in the summer of 2002. Frequent intense precipitation events throughout the summer resulted in extraordinarily wet conditions in east-central and northwestern Minnesota. Streamflow remained at record-setting high levels in many rivers and streams in these regions for several weeks. We observed high concentrations of MeHg (>1.4 ng/L) accompanied by high MeHg/THg ratios (0.39 to 0.50) in the Roseau River in northwestern Minnesota and in the Elk and Rum Rivers in east-central Minnesota. Very high MeHg mass loadings were observed in the Mississippi River just upstream of Minneapolis on July 17 (51 g MeHg/day) and July 23 (42 g MeHg/day), when MeHg concentrations at this site were 0.89 and 0.99 ng/L, respectively. The elevated MeHg concentrations in the Roseau River were associated with low dissolved oxygen and high dissolved reactive phosphorus concentrations, both of which are characteristic of anoxic waters. These rivers drain landscapes containing varying amounts of wetlands, and some of the MeHg discharged is thought to have been flushed from anoxic wetland soils. In addition, the flooding of vast areas of normally dry land surfaces probably also resulted in increased MeHg production, adding to the quantities of MeHg exported from these watersheds. Changing climate patterns are expected to result in more frequent heavy precipitation and flooding events in Minnesota. Our results suggest that as flooding and wet conditions in this region increase, the production of MeHg and its export from terrestrial areas to surface waters will increase also. PMID- 16257038 TI - Responses of Hexaplex (Murex) trunculus to selected pollutants. AB - Cadmium, copper and zinc concentrations (in whole soft body and in tissues) were measured in Hexaplex trunculus collected from the Bizerta lagoon in Tunisia. An evaluation of the biological effects of the most toxic metals (cadmium and copper) and of two organics (carbofuran and lindane), present in the sediments of the Bizerta lagoon, was attempted by measuring biomarkers (acetylcholinesterase: AChE, catalase: CAT and glutathione S-transferase: GST activities) in animals experimentally exposed for 48 or 72 h. The concentration ranges as follows: Zn>Cu>Cd. Copper concentrations are highly variable (8.0 to 235 microg g(-1) d.w.) whereas cadmium (range 1.35-4.86 microg g(-1)) and zinc (range 360-1320 microg g(-1)) concentrations are less variable. The digestive gland and the gill take up more metal than the muscle. AChE activity in H. trunculus is decreased by exposure to carbofuran or the mixture carbofuran and cadmium, in the digestive gland and muscle and by copper and by lindane in the digestive gland. AChE is generally inhibited by carbamates but some other compounds may also decrease this activity as observed in this paper. An increase in CAT activity associated with a decrease in GST activity is noted in the muscle of H. trunculus exposed to cadmium, to carbofuran and to the mixture of cadmium and carbofuran, and in the digestive gland of animals exposed to lindane. These pollutants may act upon glutathione and decrease the GST activity that cannot detoxify them and CAT activity has a protective effect. On the contrary, copper increases CAT and GST activities in the digestive gland of exposed gastropods; these enzymes seem to cooperate and play together their role of anti-oxidant enzymes. If H. trunculus is not a bioindicator species for metal concentrations, due to a high variability in metal concentrations, nevertheless the biochemical responses to pollutants (cadmium, copper, carbofuran and lindane) represented by AChE, CAT and GST activities may act as biomarkers of exposure in this species. PMID- 16257040 TI - Transparent exopolymeric particles' distribution in the northern Adriatic and their relation to microphytoplankton biomass and composition. AB - Transparent exopolymer particles (TEP) are generated during transformation processes within the non-living organic matter continuum in the seawater. Seasonal dynamics of TEP along the Po River Delta-Rovinj transect was followed during the 3-year period, using a spectrophotometric method of determination of TEP. Relationships between TEP and phytoplankton biomass (chlorophyll a), abundance and composition, as well as phaeophytin, were analysed. Microphytoplankton spring blooms are confirmed to be an important source of TEP, with some species more related to TEP concentration dynamics, such as Chaetoceros sp. and Skeletonema costatum. Notable spatial west to east gradient of autotrophic biomass and consequently of TEP was observed, as well as gradual decrease with depth. In addition, TEP concentration increments were observed up to 2 months before the mucilage events followed by the decrease of TEP concentration during the event itself. Pattern of TEP dynamics in the years with mucilage events was the most pronounced in 2002, when the event was spatially and temporally the most extended. PMID- 16257042 TI - Neoadjuvant chemotherapy in vulvar cancer: avoiding primary exenteration. AB - OBJECTIVE: To determine whether neoadjuvant cisplatin and 5-fluorouracil chemotherapy can be used to preserve the anal sphincter and/or urethra in patients with advanced vulvar cancer involving these sites. METHODS: Fourteen patients with advanced vulvar cancer (1997-2003) involving the anal sphincter and/or urethra were given 3-4 cycles of neoadjuvant chemotherapy to attempt preservation of these pelvic structures rather than undergoing a primary pelvic exenteration. Following 3 cycles, a radical vulvectomy and groin lymph node dissection were planned. All patients had lesion size documented by measurement and photograph prior to and following chemotherapy. RESULTS: The median age was 63 years (range 39-88). Thirteen patients received a median of 3 cycles (range 2 4) of neoadjuvant chemotherapy. Ten patients received cisplatin and 5 fluorouracil, while three received cisplatin alone. The median time from diagnosis to surgery was 77 days (range 54-143). All patients with cisplatin and 5-fluorouracil chemotherapy underwent surgery except one patient who had a synchronous renal cell carcinoma and died prior to surgery. Patients receiving cisplatin alone showed no measurable response, while all patients receiving cisplatin and 5-fluorouracil demonstrated at least a partial response. Two patients had no residual invasive carcinoma on final pathology. All patients receiving cisplatin and 5-fluorouracil followed by surgery are disease-free, while two of three receiving cisplatin have progressive disease. The anal sphincter and urethra were conserved in all patients receiving cisplatin and 5 fluorouracil. CONCLUSION: Neoadjuvant cisplatin and 5-fluorouracil in advanced vulvar cancer demonstrated a response rate of 100%. The anal sphincter and urethra were conserved in all patients receiving cisplatin and 5-fluorouracil. Responders are disease-free at this time. This response rate demonstrates superior activity of 5-fluorouracil in vulvar cancer and spares these patients the morbidity of exenteration or radiation. PMID- 16257043 TI - Durable response of metastatic endometrial carcinoma to treatment with fulvestrant (Faslodex) after prior progestin and anastrozole therapy. AB - BACKGROUND: Patients with metastatic endometrial carcinoma may respond to hormonal therapy with progestins. There is a need for new therapies for hormone responsive disease. CASE: We report a patient with metastatic endometrial carcinoma to the lungs, who after progressing on progestin therapy, had a lengthy remission with anastrozole; upon further progression, fulvestrant (Faslodex) was instituted, with a resultant partial remission, which has been sustained for almost 3 years. CONCLUSION: In this case, fulvestrant therapy was successful even in the face of prior anastrozole and megestrol. The activity of fulvestrant in patients with metastatic endometrial carcinoma should be further explored, especially in situations in which the tumor is well differentiated and/or expresses hormone receptors. PMID- 16257044 TI - Ovarian cancer presenting as chest wall subcutaneous nodule: a case report and a literature review. AB - BACKGROUND: Distant metastasis from primary epithelial ovarian carcinoma is commonly found as nodal and intraperitoneal spread, spread via haematogenous routes or transcoelomic spread. Ovarian cancer presents in different ways, but common presentations include abdominal pain, distention or ascites due to metastatic involvement of peritoneal cavity. Most tumour present at advanced stage and distant metastases to common and uncommon sites are found in patients who have undergone treatment for primary ovarian cancer. Subcutaneous metastatic nodules from primary ovarian cancer are rarely found in advanced disease. We describe a case of asymptomatic ovarian carcinoma presenting as a chest wall nodule. CASE: An unusual case of primary ovarian carcinoma presenting as asymptomatic chest wall subcutaneous nodules that subsequently were diagnosed as metastatic lesions. CONCLUSION: An unusual case of ovarian carcinoma where the patient was totally asymptomatic and referred with two tiny subcutaneous nodules. Therefore, lumps of recent onset, although asymptomatic, should either have fine needle aspiration cytology or excision biopsy. PMID- 16257045 TI - Increased palatability of ethanol after prenatal ethanol exposure is mediated by the opioid system. AB - Previous studies have shown that prenatal exposure to a moderate dose of ethanol (2 g/kg) during the last days of gestation of the rat (17-20) not only increases postnatal intake of the drug but also enhances the palatability of ethanol's taste when measured with a taste reactivity test. Prenatal administration of the opioid antagonist naloxone, together with ethanol, reduces ethanol intake. The aim of the present study was to analyze whether this decreased intake of ethanol after the administration of naloxone is accompanied by a reduction in ethanol's palatability. Results show that preweanling rats exposed prenatally to ethanol alone displayed more ethanol intake and more ingestive responses in reaction to its taste than non-exposed pups. Simultaneous prenatal administration of naloxone with ethanol prevented both the increased intake of ethanol and the higher amount of appetitive responses to its taste. These results indicate that the opioid system plays an important role in the effect of enhanced palatability of ethanol's taste after its prenatal exposure. Results also support the hypothesis of a conditioned response established in utero as a consequence of the association between ethanol's chemosensory and reinforcing aspects, the latter mediated by the opioid system. PMID- 16257046 TI - Hypertension prior to athletic events: are we heading for a disaster? PMID- 16257047 TI - Correlates of condom failure among adolescent males: an exploratory study. AB - OBJECTIVE: To identify the prevalence and correlates of condom failure (defined as breakage or slipping off in the past 90 days) among a sample of adolescent males (15 to 21 years of age). DESIGN: A cross-sectional study of 481 condom using males residing in three US cities (Atlanta, GA, Providence RI, Miami FL). Data were collected, in the years 2000 and 2001, using audio computer-assisted self-interviewing technology. Prevalence ratios were used to determine the strength and significance of bivariate associations between ten assessed correlates and condom failure. Correlates achieving a screening level of significance were entered into a multivariate model that was used to calculate adjusted odds ratios (AOR). RESULTS: Recent condom failure was reported by 34.1%. Younger adolescents were about one-third less likely to report condom failure (AOR = 0.66; P = 0.4). Adolescents reporting multiple sex partners were about 80% more likely to report failure (AOR = 1.84; P = 0.09). Adolescents indicating they had sex with someone on the same day they met the person were about 80% more likely to report failure (AOR = 1.77; P = 0.02). Finally, adolescents indicating recent problems obtaining condoms were about 70% more likely to report failure (AOR = 1.69; P = 0.1). Failure was not less common among those reporting a history of STD infection or those ever impregnating a partner. CONCLUSION: Because adolescent males may commonly experience condom failure, targeted clinic- and community-based programs designed to reduce user error could be an important aspect of preventing pregnancy and the spread of STDs. PMID- 16257048 TI - Effect of organic and inorganic acids on concentrated chitosan solutions and gels. AB - Rheological properties of concentrated chitosan aqueous solutions and gels in the presence of different organic and inorganic acids were investigated. Viscosities of the solutions increased with polymer concentration and degree of ionization. Strong gels were obtained at pH around 2 with oxalic, phosphoric and sulfuric acids. Gelation was favored by simple and short chain length acids and was governed by ionic interactions. The gels could be distinguished from solutions by the frequency independence of their dynamic moduli and their high apparent activation energy for flow. PMID- 16257049 TI - Mechanical compression affecting the thermal-induced conformational stability and denaturation temperature of human fibrinogen. AB - Thermal-induced conformational stability and changes in denaturation temperature of human fibrinogen (FBG) after different mechanical compressions were investigated by a simultaneous Fourier transform infrared microspectroscopy equipped with thermal analyzer (thermal FTIR microscopic system). The confocal Raman microspectroscopy was also applied to determine the thermal reversibility of solid FBG. FBG powder was pressed on one KBr pellet (1 KBr method) or sealed within two KBr pellets (2 KBr method) by different mechanical compressions. The result indicates that there was no marked difference in the thermal behavior for the solid FBG samples prepared by 1 KBr method in the heating process even under different mechanical compression pressures, in which the thermal-induced denaturation temperatures from native to denatured state were maintained constant at 66-67 degrees C. However, the denaturation temperature for the solid FBG samples prepared by 2 KBr method was shifted from 55 to 62 degrees C with the increase of mechanical compression pressure. A good linear correlation was also found between the denaturation temperature and mechanical compression pressure for FBG samples prepared by 2 KBr method. The solid FBG sample, whether prepared by 1 KBr or 2 KBr method, was also found to show the thermal-irreversible property. PMID- 16257050 TI - Formation of bone-like apatite on organic polymers treated with a silane-coupling agent and a titania solution. AB - Polyethylene terephthalate (PET), ethylene-vinyl alcohol copolymer (EVOH) and Nylon 6 in plate form were treated with silane-coupling agents, a titanium alkoxide-alcohol solution and a hot HCl solution to form a thin crystalline titanium oxide layer. When placed in a simulated body fluid with ion concentrations nearly equal to those of the human blood plasma, nanosized bone like apatite formed uniformly on the surfaces of these treated polymers: within 2 days for PET and Nylon 6, and 7 days for EVOH. This indicates that such titania modified polymers might form bone-like apatite in the living body, and bond to living bone through this apatite layer. Three-dimensional fabrics of these polymer fibers, with open spaces in various sizes, are expected to be useful as bone substitutes, as they will be integrated with the natural bone. PMID- 16257051 TI - Modelling approach in cell/material interactions studies. AB - Based on our experiments, we propose a statistical modeling approach of the in vitro interactions between biological objects and materials. The objective of this paper is to provide basic principles for developing more ambitious experiments comparing the simultaneous influence of more than one or two parameters on various observations, taking advantage of convenient statistical and mathematical techniques for the treatment of measured data. Analyzing some examples of our own experiments, the essential features needed for modeling cell/material interaction studies are presented. Firstly, we describe the initial process of designing appropriate experiments that allow for comprehensive modeling. In the second part, we illustrate the different applications of a specific statistical modeling technique, the bootstrap protocol, on either the amplification of data, the elimination of correlation existing between measured parameters or, out of a set of parameters, identification of the most relevant parameter for further statistical analysis. Finally, based on recent statistical analysis tools such as the bootstrap, we illustrate the relative influence of biological and physical parameters in phenomenological studies of cell/material interactions. PMID- 16257052 TI - hCG Secretion in human choriocarcinoma JAR cells is MAPK but not Stat3 dependent: contributions of TNFalpha and IL-1beta to inflammation-induced hCG secretion. AB - Inflammatory mediators play critical physiologic roles throughout the course of normal pregnancy. At the same time, uncontrolled inflammation appears to have an adverse affect on pregnancy outcome. Thus, a crucial task of reproductive immunology is to define the mechanisms through which inflammation is controlled at the maternal-fetal interface. We examined the signaling pathways activated in the human choriocarcinoma cell line, JAR, in response to an in vitro model of an inflammatory challenge. We incubated JAR cells with medium from peripheral blood mononuclear cells (PBMC) that had been activated with either LPS or PHA. Conditioned medium from each experimental model induced MAP kinase and Stat3 phosphorylation as well as human chorionic gonadotropin (hCG) secretion from JAR cells. hCG secretion could be blocked by pharmacologic inhibition of MAP kinase but not by inhibition of Stat3 using an siRNA approach. The MAPK activators IL 1beta and TNFalpha both induced hCG secretion from JAR cells, but not the Stat3 activators IFN-gamma and IL-6. Furthermore, hCG secretion induced by conditioned media could be blocked by IL-1 receptor antagonist. We conclude that inflammation at the maternal-fetal interface may involve complex webs of regulatory and counter-regulatory mechanisms that involve multiple cytokines and at least two major, independent cell-signaling systems. PMID- 16257053 TI - Complete nucleotide sequence of pSCV50, the virulence plasmid of Salmonella enterica serovar Choleraesuis SC-B67. AB - We carried out comparative analysis on the sequences of two 50-kb virulence plasmids of Salmonella enterica serovar Choleraesuis strains SC-B67 (pSCV50) and RF-1 (pKDSC50). The two plasmids share over 99% sequence similarity. Ninety-two nucleotide variations at 42 sites were detected between the two plasmids; pSCV50 contains 24 nucleotide substitutions, 6 deletions, and 62 insertions, compared to pKDSC50. Two regions in pSCV50 appeared to be more susceptible to changes: one is the non-virulence-associated transfer region (27.5-33.0 K) and the other a function-unknown region (9.0-10.5 K). We re-annotated pSCV50 using more advanced tools and the up-to-date databases and corrected the inaccurate annotation in pKDSC50. The results indicate that virulence-related genes on the 50-kb plasmid are under negative selection, suggesting that they play important roles in the expression of virulence during the process of infection, while other genes in this plasmid tend to evolve neutrally. PMID- 16257054 TI - Direct and noninvasive assessment of parafoveal capillary leukocyte velocity. AB - PURPOSE: Alterations in leukocyte velocity have been implicated in many retinal disease processes. However, direct and objective assessment of leukocyte velocity in retinal capillaries has been limited by a reliance on invasive contrast dyes that allow leukocyte visualization only for a short time span. The recent application of adaptive optics in a scanning laser ophthalmoscope (AOSLO) has made long-term imaging of parafoveal leukocyte movement possible without contrast dyes. In this study, using the AOSLO, we demonstrate a new method to investigate retinal parafoveal capillary leukocyte velocity. DESIGN: Experimental study. PARTICIPANTS: Six normal healthy subjects ranging from 25 to 35 years of age with clear ocular media. METHODS: The parafoveal zone of the retina was imaged in all subjects using an AOSLO. MAIN OUTCOME MEASURES: Leukocyte velocity was determined in the parafoveal capillaries including the foveal avascular zone border. Leukocyte velocity was measured directly from movie segments in which the leukocytes were clearly visible. RESULTS: The mean parafoveal leukocyte velocity for 6 subjects was 1.37 mm/second, ranging from 0.77 to 2.10 mm/second. Leukocytes were not visible in all parafoveal capillaries. CONCLUSIONS: Parafoveal capillary leukocyte velocity can be directly and noninvasively measured without the use of contrast dyes using an AOSLO. PMID- 16257055 TI - BCL-XL expression levels influence differential regional astrocytic susceptibility to 1,3-dinitrobenzene. AB - The selective vulnerability of brainstem astrocytes to 1,3-dinitrobenzene is mediated by a 10-fold lower threshold for opening of the cyclosporin A inhibitable mitochondrial permeability transition pore (mtPTP). BCL-XL, BAX and BCL-2 are members of the BCL-2 protein family known to regulate both apoptotic and necrotic cell death signaling at the mtPTP. The levels at which these proteins are expressed relative to one another, where in the cell they are located and whether they are post-translational modified contributes greatly to the balance in active agonistic to active antagonistic BCL-2 proteins, and this critical balance has been hypothesized to dictate regional astrocytic susceptibility to DNB. The effects of DNB on the balance in expression of the BCL 2 family proteins have been evaluated in F344 rat DNB-sensitive (brainstem) and non-sensitive (cortical) tissue homogenates and primary astrocytes. No significant treatment-related alterations in BCL-XL, BAX or BCL-2 protein expression are observed in rat tissue homogenates or primary astrocytes. However, moderate increases in BCL-XL are observed only in DNB-treated rat cortical astrocytes, and these increases may be sufficient to shift the constitutive balance in expression of antagonistic to agonistic BCL-2 proteins from a ratio which favors BAX to one in which BAX and BCL-XL are comparably expressed. Rat primary brainstem and cortical astrocytes are also transiently transfected with bcl-xl to evaluate whether or not moderate enhancement of BCL-XL protein expression levels are sufficient to alter regional sensitivity to DNB in vitro. BCL-XL overexpression minimizes DNB-induced inhibition of succinate dehydrogenase (complex II) activity and increases significantly the concentration of DNB required to induce MPT onset in primary brainstem and cortical astrocytes. Results from the current investigation suggest that modest region-specific alterations in the balance in expression of antagonistic to agonistic BCL-2 proteins may adequately explain differential regional sensitivity to DNB-induced mitochondrial dysfunction. PMID- 16257056 TI - High-speed gas sensor for chemosensory event-related potentials or magnetic fields. AB - The observation of odor and air exchange with high temporal accuracy is necessary to obtain strict chemosensory event-related potentials (CSERPs) or magnetic fields, as proposed by Evans et al. [Evans W, Kobal G, Lorig T, Prah J. Suggestions for collection and reporting of chemosensory (olfactory) event related potentials. Chem Senses, 1993; 18: 751- 6]. No suitable method for real time observation of gas stimuli, however, has been available until now. We have developed a technique to measure accurately gas molecule concentrations with a high temporal resolution. We determined that attenuation of sound amplitude varies in a manner dependent on the average molecular weight through which the sound wave passes. Based on this principle, we have designed a high-speed gas concentration sensor utilizing ultrasound. We investigated the practical potential of this sensor using a chemosensory stimulator (olfactometer); we succeeded in observing rapid gas exchange between air and nitrogen with a 2 kHz sampling rate. The signal/noise ratio of the stimulus was greater than 42 dB. In a 20 min experiment we determined that, for this olfactometer, the gas onset latency was 79 ms and the rise time was 16 ms. No significant artifact to magnetic fields was observed, even when the sensor was situated near a whole head magnetoencephalography (MEG) system. These results indicate that this sensor could be used for the observation of odor and air exchange, as well as, for real time monitoring of odor stimuli during actual experiments with a participant. PMID- 16257057 TI - A long-range, wide field-of-view infrared eyeblink detector. AB - Classical conditioning of the eyeblink response in the rabbit is one of the most advanced models of learning and memory in the mammalian brain. Successful use of the eyeblink conditioning paradigm requires precise measurements of the eyeblink response. One common technique of eyelid movement detection utilizes measurements of infrared (IR) light reflected from the surface of the eye. The performance of current IR sensors, however, is limited by their sensitivity to ambient infrared noise, by their small field-of-view and by short working distances. To address these limitations, we developed an IR eyeblink detector consisting of a pulsing (62.5 kHz) IR light emitting diode (LED) paired with a silicon IR photodiode and circuit that synchronously demodulates the recorded signal and rejects background IR noise. The working distance of the sensor exceeds 20 mm, and the field-of-view is larger than the area of a rabbit's eye. Due to its superior characteristics, the new sensor is ideally suited for both standard eyeblink conditioning and for studies that utilize IR-containing visual stimuli and/or that are conducted in an environment contaminated with IR noise. PMID- 16257058 TI - Exposure of the rat tail to ultraviolet A light produces sustained hyperalgesia to noxious thermal and mechanical challenges. AB - We aimed to establish whether exposing the tails of rats to ultraviolet A (UVA) light generated sustained hyperalgesia to noxious thermal and mechanical challenges. The tails of 21 rats underwent eight 40s exposures of UVA light, with 260s between each exposure. As a control procedure, during UVA-light exposure the tails of 11 of those rats were shielded with aluminium foil. Thermal hyperalgesia was assessed by immersing the rat tail in 49 degrees C water (modified tail flick test). Mechanical hyperalgesia was assessed by applying a bar algometer onto the tail and timing the escape response. Exposure to direct UVA light produced hyperalgesia for 8 days to the noxious thermal challenge (P<0.05, two-way ANOVA, Tukey post hoc tests) and at least 16 days to the noxious mechanical challenge (P<0.05, two-way ANOVA, Tukey post hoc tests). They gained mass throughout the study at the same rate as the control rats. The control rats did not develop thermal nor mechanical hyperalgesia. The tails of a further 20 rats were exposed similarly, and tail tissue examined histologically. Both exposed and control rats developed mild chronic inflammation unrelated to the hyperalgesia. PMID- 16257059 TI - Does power matter with ECT? AB - BACKGROUND: To assess the incidence of depressed inpatients requiring high output ECT and the response of this group compared with a group requiring standard output ECT. METHODS: We reviewed the records of 59 consecutive inpatients that were treated with bilateral ECT between January 2001 and January 2004. Diagnosis of major depression was based on DSM IV criteria. Response and remission to ECT (respectively defined as a 50% reduction in score and a score of < or = 7 on the Hamilton Rating Scale for Depression; HRSD) of both groups were compared. RESULTS: Of the 59 patients, 13 (22%) required high output ECT. These patients needed significantly more ECT treatments than patients in the standard dose group (16.4+/-7.1 versus 10.4+/-4.5; p=0.01). In total, 31 of 46 patients (67%) requiring standard output ECT and 11 of the 13 patients (85%) requiring high output ECT responded to ECT. This difference is not significant. LIMITATIONS: This study has a retrospective nature and a rather homogenous sample. CONCLUSION: In this study 1 in 5 of the depressed inpatients needs a high dose energy of bilateral ECT to induce an adequate seizure. The efficacy of ECT in these patients is similar to that in the standard dose group. Considering these facts, high output ECT devices should be available for use in routine clinical practice. PMID- 16257060 TI - Administration of MIP-3alpha gene to the tumor following radiation therapy boosts anti-tumor immunity in a murine model of lung carcinoma. AB - Dendritic cells (DC), the most potent antigen presenting cells (APC), have been shown able to process apoptotic tumor cells and necrotic tumor cells for antigen presentation. Apoptosis and necrosis are the two common final pathways through which the tumors are killed by chemotherapy or radiation therapy. The tumor cells receiving radiation often produce the "danger signal" cytokines such as TNF-alpha and IL-1. Another cytokine MIP-3alpha that is able to attract DC to the tumor site is normally not secreted. We hypothesize that if artificial introduction of a large number of DC to the necrotic tumor site after radiation therapy by transfecting any cells at the tumor site to secrete DC-tropic MIP-3alpha, an anti tumor immune response would be initiated. C57BL/6J mice bearing a well-known Lewis lung carcinoma are used to assess efficacy of this modality. The plasmid DNA containing pcDNA3.1/MIP-3alpha was injected into the subcutaneous tumors after radiation treatment. We demonstrate a detectable local expression of MIP 3alpha and local accumulation of DC. Tumor infiltrating lymphocytes after the treatment are predominantly CD8+ T-cells with rare CD4+ T-cells. The anti-tumor immune response was also measurable, which contributes at least in part to the finding that the treated mice have smaller tumor and prolonged survival, comparing to the control groups. This study suggests a potential new means of immune modulation and provides us a new concept of immunotherapy of cancer. PMID- 16257061 TI - The cytokine signature of MOG-specific CD4 cells in the EAE of C57BL/6 mice. AB - Experimental allergic encephalomyelitis (EAE) is an animal model of multiple sclerosis. While EAE is mediated by the cytokines produced by specific T cells, the cytokine signature of these effector cells is unresolved. We tested CD4 cells from MOG peptide 35-55 immunized C57BL/6 mice for their peptide induced cytokine production on antigen presenting cells of the respective cytokine knockout mice, or wild type mice. IL-4 and IL-6 production was seen on wild type antigen presenting cells, suggesting that IL-4 and IL-6 are not T cell products. In contrast, IFN-gamma, IL-2 and IL-3 were found to be produced by the MOG specific CD4 cells. Understanding the cognate vs. bystander cytokine production in EAE might help dissect the contribution of cytokines to the pathogenesis of the disease. PMID- 16257062 TI - Activation of adenosine A(1) receptors alters behavioral and biochemical parameters in hyperthyroid rats. AB - Adenosine acting on A(1) receptors has been related with neuroprotective and neuromodulatory actions, protection against oxidative stress and decrease of anxiety and nociceptive signaling. Previous studies demonstrated an inhibition of the enzymes that hydrolyze ATP to adenosine in the rat central nervous system after hyperthyroidism induction. Manifestations of hyperthyroidism include increased anxiety, nervousness, high O(2) consumption and physical hyperactivity. Here, we investigated the effects of administration of a specific agonist of adenosine A(1) receptor (N(6)-cyclopentyladenosine; CPA) on nociception, anxiety, exploratory response, locomotion and brain oxidative stress of hyperthyroid rats. Hyperthyroidism was induced by daily intraperitoneal injections of l-thyroxine (T4) for 14 days. Nociception was assessed with a tail-flick apparatus and exploratory behavior, locomotion and anxiety were analyzed by open-field and plus maze tests. We verified the total antioxidant reactivity (TAR), lipid peroxide levels by the thiobarbituric acid reactive species (TBARS) reaction and the free radicals content by the DCF test. Our results demonstrated that CPA reverted the hyperalgesia induced by hyperthyroidism and decreased the exploratory behavior, locomotion and anxiety in hyperthyroid rats. Furthermore, CPA decreased lipid peroxidation in hippocampus and cerebral cortex of control rats and in cerebral cortex of hyperthyroid rats. CPA also increased the total antioxidant reactivity in hippocampus and cerebral cortex of control and hyperthyroid rats, but the production of free radicals verified by the DCF test was changed only in cerebral cortex. These results suggest that some of the hyperthyroidism effects are subjected to regulation by adenosine A(1) receptor, demonstrating the involvement of the adenosinergic system in this pathology. PMID- 16257063 TI - Ecotoxicology of human pharmaceuticals. AB - Low levels of human medicines (pharmaceuticals) have been detected in many countries in sewage treatment plant (STP) effluents, surface waters, seawaters, groundwater and some drinking waters. For some pharmaceuticals effects on aquatic organisms have been investigated in acute toxicity assays. The chronic toxicity and potential subtle effects are only marginally known, however. Here, we critically review the current knowledge about human pharmaceuticals in the environment and address several key questions. What kind of pharmaceuticals and what concentrations occur in the aquatic environment? What is the fate in surface water and in STP? What are the modes of action of these compounds in humans and are there similar targets in lower animals? What acute and chronic ecotoxicological effects may be elicited by pharmaceuticals and by mixtures? What are the effect concentrations and how do they relate to environmental levels? Our review shows that only very little is known about long-term effects of pharmaceuticals to aquatic organisms, in particular with respect to biological targets. For most human medicines analyzed, acute effects to aquatic organisms are unlikely, except for spills. For investigated pharmaceuticals chronic lowest observed effect concentrations (LOEC) in standard laboratory organisms are about two orders of magnitude higher than maximal concentrations in STP effluents. For diclofenac, the LOEC for fish toxicity was in the range of wastewater concentrations, whereas the LOEC of propranolol and fluoxetine for zooplankton and benthic organisms were near to maximal measured STP effluent concentrations. In surface water, concentrations are lower and so are the environmental risks. However, targeted ecotoxicological studies are lacking almost entirely and such investigations are needed focusing on subtle environmental effects. This will allow better and comprehensive risk assessments of pharmaceuticals in the future. PMID- 16257064 TI - Cloning and characterization of angiotensin converting enzyme related dipeptidylcarboxypeptidase from Leishmania donovani. AB - We report the first identification, gene cloning, recombinant expression and biochemical characterization of an angiotensin converting enzyme (ACE) related dipeptidylcarboxypeptidase (DCP) in a protozoan parasite. The mammalian counterpart of this enzyme, peptidyl dipeptidase A (a carboxyl dipeptidase) also known as ACE leads to the cleavage of angiotensin I to produce a potent vasopressor. The catalytic enzyme activity of its Escherichia coli DCP counter part can be inhibited by the antihypertensive drug captopril, suggesting that this class of enzymes constitutes a novel target for drugs and vaccines. By utilizing a DNA microarray expression profiling approach, we identified a gene encoding a DCP enzyme for the kinetoplast protozoan Leishmania donovani (LdDCP) that was differentially expressed in promastigote and amastigote stages of the parasite life cycle. Both RNA and protein levels of LdDCP are higher in axenic amastigotes compared to promastigotes. Immuno-fluorescence analysis revealed the cytosolic expression of the protein. Primary structure analysis of LdDCP revealed the presence of an active Zn binding site. When expressed in E. coli, the recombinant enzyme showed carboxy-dipeptidase activity with synthetic substrates. Replacement of two histidine and one glutamic acid at positions 466, 470 and 467, respectively, with alanine residues in its active site resulted in loss of enzyme activity. Captopril, an ACE specific inhibitor was able both to reduce significantly LdDCP enzyme activity and to inhibit promastigote growth. Both its cytosolic location and close homology to DCPs from bacterial species suggests a role in parasite nutrition. Further, identification of LdDCP now provides an opportunity to investigate Leishmania peptidases for their potential as drug and vaccine targets. PMID- 16257065 TI - Evolution of non-LTR retrotransposons in the trypanosomatid genomes: Leishmania major has lost the active elements. AB - The ingi and L1Tc non-LTR retrotransposons--which constitute the ingi clade--are abundant in the genome of the trypanosomatid species Trypanosoma brucei and Trypanosoma cruzi, respectively. The corresponding retroelements, however, are not present in the genome of a closely related trypanosomatid, Leishmania major. To study the evolution of non-LTR retrotransposons in trypanosomatids, we have analyzed all ingi/L1Tc elements and highly degenerate ingi/L1Tc-related sequences identified in the recently completed T. brucei, T. cruzi and L. major genomes. The coding sequences of 242 degenerate ingi/L1Tc-related elements (DIREs) in all three genomes were reconstituted by removing the numerous frame shifts. Three independent phylogenetic analyses conducted on the conserved domains encoded by these elements show that all DIREs, including the 52 L. major DIREs, form a monophyletic group belonging to the ingi clade. This indicates that the trypanosomatid ancestor contained active mobile elements that have been retained in the Trypanosoma species, but were lost from L. major genome, where only remnants (DIRE) are detectable. All 242 DIREs analyzed group together according to their species origin with the exception of 11 T. cruzi DIREs which are close to the T. brucei ingi/DIRE families. Considering the absence of known horizontal transfer between the African T. brucei and the South-American T. cruzi, this suggests that this group of elements evolved at a lower rate when compared to the other trypanosomatid elements. Interestingly, the only nucleotide sequence conserved between ingi and L1Tc (the first 79 residues) is also present at the 5' extremity of all the full length DIREs and suggests a possible role for this conserved motif, as well as for DIREs. PMID- 16257066 TI - Characterization of a half-size ATP-binding cassette transporter gene which may be a useful marker for ivermectin selection in Onchocerca volvulus. AB - ATP-binding cassette (ABC) transporters comprise a large paralogous protein family and several confer drug resistance. Ivermectin (IVM) is the only drug approved for treatment of onchocerciasis and is a substrate for some ABC transporters. Furthermore, there is accumulating evidence that IVM selects on some ABC transporter genes in Onchocerca volvulus and other parasitic nematodes. The onchocerciasis control programs rely on community based treatment with IVM each year to reduce morbidity and decrease parasite transmission. This appears to be imposing selection pressure on O. volvulus. A half-size ABC transporter cDNA has previously been reported for O. volvulus, however, the full length gene has not been previously characterized and investigated for possible selection by IVM. Genes under selection may be identified by patterns of linkage disequilibrium (LD) and a loss of genetic polymorphism at physically linked loci. Twelve genetic markers spanning the full gene were examined in O. volvulus from non-treated and IVM treated populations in Ghana. Analysis of the genomic organization of the half-size ABC transporter (OvPLP) indicates that it spans approximately 3.8 kb comprising nine exons. Worms from treated people showed a reduction in gene diversity, a loss of genetic polymorphism at several markers, a selection for specific alleles, and a reduction in the number of regions in LD; these effects were more pronounced as treatment increased. These changes suggest that IVM is imposing selection pressure on this gene. The region between transmembrane domains four and five may be a useful marker for IVM selection in O. volvulus. PMID- 16257067 TI - Making sense of pragmatic criteria for the selection of geriatric rehabilitation measurement tools. AB - In geriatric rehabilitation, the selection of the most appropriate measurement tools involves pragmatic as well as psychometric considerations. However, there is no consensus about the conceptual and operational definitions of the pragmatic criteria involved in this selection. The objective of this research was to identify such operational criteria through a literature search between 1995 and 2004. Results identified operational criteria that were grouped under four categories using a conceptual mapping methodology: respondent burden, examiner burden, score distribution and format compatibility. We recommend the umbrella term applicability to refer to this grouping of pragmatic qualities of a measurement tool. Examining the applicability of measurement tools should assist clinicians and researchers in selecting the most appropriate for use in geriatric rehabilitation. PMID- 16257068 TI - Iliac artery thrombosis after aortic balloon counterpulsation: treatment with intraarterial tirofiban, manual thrombectomy and stenting. PMID- 16257069 TI - Electric shock: Cardiac effects relative to non fatal injuries and post-mortem findings in fatal cases. AB - The documented cardiovascular effects of an electrical shock include acute myocardial necrosis, myocardial ischemia with or without necrosis, heart failure, arrhythmias, haemorrhagic pericarditis, acute hypertension with peripheral vasospasm and anomalous, non specific ECG alterations. Studies documenting the development of acute left ventricular failure and pulmonary oedema sustain that the observed ECG changes are secondary to myocardial injury. The cause of death of victims of instantly fatal electrical accidents is ventricular fibrillation, but it is not clear if this fibrillation is due to purely electrophysiological changes or to identifiable structural abnormalities in the heart. Little is known about the morphological changes in the heart, as differing anatomical alterations are described. Data from such studies may aid in a more accurate comprehension of clinical and morphological alterations of the heart and in the development of therapeutic strategies that could counterbalance such effects. PMID- 16257070 TI - Myocardial expression of survivin, an apoptosis inhibitor, in aging and heart failure. An experimental study in the spontaneously hypertensive rat. AB - BACKGROUND: Apoptosis plays a major role in the transition to heart failure (HF) in systemic hypertension although the underlying mechanisms are still unclear. The aim of this study was to determine the relationship between apoptosis, left ventricular remodeling, heart failure and the myocyte expression of survivin, an inhibitor of apoptosis. METHODS: Spontaneously hypertensive rats (SHR) were used as a model of hypertensive cardiopathy, and Wistar Kyoto Stars rats (WKY) were used as controls. Animals were allowed to survive up to 18 months of age. The animals underwent echocardiography (EDD, ESD and FS were measured). The median section of the heart was processed for in situ end-labeling of DNA fragmentation (TUNEL) and for survivin expression by immunohistochemistry. RESULTS: All SHR presented features of adverse cardiac remodeling. Apoptotic cells were increased in SHR compared with WKY, measured as apoptotic cells per high power field (1.08+/-0.43 vs. 0.27+/-0.15, P<0.001), and as apoptotic rate (0.16+/-0.06% vs. 0.04+/-0.02%, P<0.001). The incidence of apoptosis showed a positive correlation with unfavorable ventricular remodeling, assessed by echocardiogram. Survivin expression was found in all cases, but the survivin expression index was significantly lower in SHR vs. WKY (43+/-40% vs. 86+/-18%, respectively, P=0.014). Moreover the survivin expression index was inversely correlated with features of adverse remodeling (i.e., Heart Weight, R=-0.79, P<0.001) and with apoptosis (i.e., apoptotic rate, R=-0.52, P=0.050). CONCLUSION: Survivin myocardial expression in aging SHR is associated with reduced apoptosis and more favorable cardiac remodeling. Modulation of this pathway may prove beneficial in preventing pressure overload cardiac remodeling and heart failure. PMID- 16257071 TI - Chlamydia pneumoniae, harmful only for peripheral arteries? PMID- 16257072 TI - Typical clinical aspect of endomyocardial fibrosis. AB - Endomyocardial fibrosis is a restrictive cardiomyopathy, of unknown etiology, which occurs most commonly in tropical and subtropical areas. Typical clinical aspect of the disease shows striking ascites, regardless of which ventricle is involved, contrasting with little or no peripheral edema. PMID- 16257073 TI - Gene expression profiling of human cardiac potassium and sodium channels. AB - BACKGROUND: The native cardiac ion currents and the action potential itself are the results of the concerted action of several different ion channels. The electrophysiological properties of cardiac cells are determined by the composition of ion channels and by their absolute abundance and proportional ratio. METHODS: Our aim in this study was to compare the gene expression level of a representative panel of cardiac ion channels with each other and to compare the same channels in the atrium and ventricle of the human heart using quantitative real-time PCR analysis. RESULTS: We obtained a significant difference in the gene expression levels in 21 of 35 channels between atrium and ventricle of healthy human hearts. Further, we found that the expression levels of Kv1.5 and Kv2.1 transcripts in the ventricle were very high, and that mRNAs for Kv1.7 and Kv3.4 are highly abundant in both the atrium and ventricle, which might indicate a functional role of these ion channel subunits in the formation of action potential in the human ventricle and both in the atrium and ventricle, respectively. CONCLUSIONS: This is the first report on the expression of several ion channel subunits, such as Kv1.7, Kv3.3 or Kv3.4 in human cardiomyocytes. The expression levels of these genes are comparable with that of well known ion channel subunits. Therefore, it is reasonable to assume, that these ion channel subunits may contribute to native currents in the human myocardium. PMID- 16257074 TI - Fluctuations in pO2 in poorly and well-oxygenated spontaneous canine tumors before and during fractionated radiation therapy. AB - BACKGROUND AND PURPOSE: The spatial heterogeneity in oxygen tension (pO2) in tumor tissue has been studied extensively, whereas, the information about the temporal heterogeneity is sparse. The purpose of the present study was to search for pO2 fluctuations in untreated and irradiated spontaneous canine tumors, and to investigate whether there is a relationship between overall tumor oxygenation status and pO2 fluctuation pattern. PATIENTS AND METHODS: Six dogs scheduled for radiation therapy of head and neck cancer were included in the study. The primary tumors were irradiated with 18 fractions of 3 Gy. Eppendorf polarographic electrodes and OxyLite fluorescence probes were used to measure overall oxygenation status and pO2 fluctuation pattern, respectively. Tissue pO2 was recorded at three subsequent days prior to treatment, and immediately before radiation fraction 4, 7, and 10. RESULTS: Overall oxygenation status differed substantially among the tumors. Radiation therapy had no consistent effect on overall oxygenation status. Fluctuations in pO2 were detected in untreated as well as irradiated tumors, and independent of whether the tumors were poorly or well oxygenated. CONCLUSIONS: Fluctuations in pO2 can occur in untreated and irradiated spontaneous canine tumors. There is no correlation between pO2 fluctuation pattern and overall tumor oxygenation status. PMID- 16257076 TI - Effects of aging on slowing of motor-response generation. AB - The aim of the present study was to analyze different stages of central processing mechanisms during a choice reaction task and to evaluate their contribution to aging-related response slowing. Event-related potentials (ERPs) were recorded from two groups of subjects, young (mean 22 years) and older adults (mean 58 years), who performed a four-alternative choice-reaction task. The results showed the expected reaction time slowing in the older subjects. This behavioural slowing was not due to delays in stimulus processing (as reflected by latencies of early ERP components), or in response selection (as reflected by the onset of the lateralized readiness potential). Instead, this slowing was due to an alteration of movement-related components, particularly an amplitude enhancement and prolongation of the motor-related potential at the cortex contralateral to the responding hand. This alteration was reliable and of general nature since it was also found in a second study using a different choice reaction task with a more direct stimulus-response relation. The results suggest that the overt response requires a higher activation level in older vs. young subjects; this extra-activation needs time and hence prolongs reaction time with aging. PMID- 16257077 TI - Voluntary stimulus production enhances deviance processing in the brain. AB - Humans often get information by voluntary action. However, little is known about how stimulus processing is modulated by self-production of stimuli. In the present study, event-related brain potentials were recorded from 16 student volunteers performing an auditory three-stimulus oddball task in two conditions. In the self condition, the stimuli were triggered by participants' voluntary button presses. In the auto condition, the same stimuli were presented automatically by a computer with the same interstimulus intervals as those in the self condition. Perceptually deviant nontarget stimuli elicited a larger P3 and a larger subsequent positivity in the self condition than in the auto condition, whereas low-deviant target stimuli elicited a P3 with equally high amplitude in both conditions. The findings suggest that voluntary stimulus production enhances orienting of attention (reflected in the P3a component) and subsequent memory updating (reflected in the P3b component) for deviant stimuli, but does not affect the response to task-relevant stimuli. Voluntary action may activate the perceptual representation of its most frequent outcomes and this anticipatory activation may make deviant stimuli more salient in the context. PMID- 16257079 TI - 30 and 69 base-pair deletion variants of the Epstein-Barr virus LMP1 oncogene occurring in Tunisian nasopharyngeal carcinoma patients. PMID- 16257078 TI - Glassy state in Bacillus subtilis spores analyzed by differential scanning calorimetry. AB - Thermal properties of dried spores of Bacillus subtilis, investigated by differential scanning calorimetry (DSC), were studied. A reversible heat capacity shift ascribable to glass-rubber transition was observed at 90-115 degrees C. The transition was found to be a pressure-inhibited volume-activated event. The decoated spores and the extracted peptidoglycan material exhibited glass transition, suggesting that the cortex could be involved in the event. Furthermore, the glass transition was evident when spores were treated with strong acid, and when the isogenic strain PS578 was scanned, indicating that core integrity and core components are not involved in the occurrence of the event. These results suggest that in the dried B. subtilis spores an amorphous biomaterial, possibly the cortex peptidoglycan, is present as a glass. PMID- 16257080 TI - CANSURV: A Windows program for population-based cancer survival analysis. AB - Patient survival is one of the most important measures of cancer patient care (the diagnosis and treatment of cancer). The optimal method for monitoring the progress of patient care across the full spectrum of provider settings is through the population-based study of cancer patient survival, which is only possible using data collected by population-based cancer registries. The probability of cure, "statistical cure", is defined for a cohort of cancer patients as the percent of patients whose annual death rate equals the death rate of general cancer-free population. Mixture cure models have been widely used to model failure time data. The models provide simultaneous estimates of the proportion of the patients cured from cancer and the distribution of the failure times for the uncured patients (latency distribution). CANSURV (CAN-cer SURVival) is a Windows software fitting both the standard survival models and the cure models to population-based cancer survival data. CANSURV can analyze both cause-specific survival data and, especially, relative survival data, which is the standard measure of net survival in population-based cancer studies. It can also fit parametric (cure) survival models to the individual data. The program is available at . The colorectal cancer survival data from the Surveillance, Epidemiology and End Results (SEER) program [Surveillance, Epidemiology and End Results Program, The Portable Survival System/Mainframe Survival System, National Cancer Institute, Bethesda, 1999.] of the National Cancer Institute, NIH is used to demonstrate the use of CANSURV program. PMID- 16257081 TI - Optical transillumination tomography with tolerance against refraction mismatch. AB - Optical transillumination tomography (OT) is a laser-based imaging modality where ballistic photons are used for projection generation. Image reconstruction is therefore similar to X-ray computed tomography. This modality promises fast image acquisition, good resolution and contrast, and inexpensive instrumentation for imaging of weakly scattering objects, such as for example tissue-engineered constructs. In spite of its advantages, OT is not widely used. One reason is its sensitivity towards changes in material refractive index along the light path. Beam refraction artefacts cause areas of overestimated tissue density and blur geometric details. A spatial filter, introduced into the beam path to eliminate scattered photons, will also remove refracted photons from the projections. In the projections, zones affected by refraction can be detected by thresholding. By using algebraic reconstruction techniques (ART) in conjunction with suitable interpolation algorithms, reconstruction artefacts can be partly avoided. Reconstructions from a test image were performed. Standard filtered backprojection (FBP) showed a round mean square (RMS) deviation from the original image of 9.9. RMS deviation with refraction-tolerant ART reconstruction was 0.33 and 0.24, depending on the algorithm, compared to 0.57 (FBP) and 0.06 (ART) in a non-refracting case. In addition, modified ART reconstruction allowed detection of small geometric details that were invisible in standard reconstructions. Refraction-tolerant ART may be the key to eliminating one of the major challenges of OT. PMID- 16257082 TI - Molecular imprinted polymer sensors: implications for therapeutics. AB - A biosensor is a sensor that uses biological selectivity to limit its perception to particular key molecules and can be defined as an analytical device possessing a biological or biologically derived sensing element integrated with or associated closely with a physicochemical transducer. In the future it is likely that a number of key developments in therapeutic monitoring and intelligent drug delivery will rely on real-time feedback information in order to deliver an appropriate response. However due to issues of integration and the fragility and unreliability of the bio-molecule, biosensors are currently unable to fulfil this role. Molecular imprinted polymers are viable alternatives to both antibodies and enzymes and this review considers the current position of molecular imprinted polymer sensing. PMID- 16257083 TI - Embryonic stem cells: meeting the needs for cell therapy. AB - Cell therapy in diverse organs has bloomed for degenerative diseases over the past decade, following a previous period of development in which haematopoietic stem cells grafts in oncology were its most prominent clinical application. One main limitation that has, however, been encountered on the path for transforming pioneering successes into real therapeutics, that would be applicable to a clinically relevant number of patients, is the difficulty in accessing "therapeutic" cells, such as foetal neurons in neurodegenerative diseases, adult pancreatic beta cells in diabetes or else myoblasts in heart failure and myopathy. The future of cell therapy definitely belongs to cell banks, from which physicians would simply have to draw according to their needs. However, besides haematopoietic stem cells, for which such infrastructures begin to exist for clinical purposes (in particular from cord blood), cell banks are still up to now essentially a scientific concept. This review focuses on the possibility for human ES cells to meet both the requirements of cell banking and the needs for substitutive cell therapy. PMID- 16257084 TI - Hospital-wide prospective mandatory surveillance of invasive aspergillosis in a French teaching hospital (2000-2002). AB - A multidisciplinary working group devoted to epidemiological surveillance of invasive aspergillosis (IA) was created in January 2000 in Grenoble University Hospital. This article presents the results of a three-year IA surveillance. The multidisciplinary working group surveyed all hospitalized patients, and the mycology laboratory detected most suspected IA cases. Cases were reviewed monthly by the Aspergillosis Committee, and were classified according to international consensus criteria. Possible nosocomial acquisition was determined. Among the 490 alerts, 74 IA cases were observed: six proven cases (8%), 36 (49%) probable cases and 32 (43%) possible cases. The incidence was 4.4 (95% CI 3.4-5.4) IA/100 000 patient-days. Among the proven and probable IA cases, we observed 10 nosocomial cases and six cases of undetermined origin. No cases were noted in the protected rooms in the haematology unit. Only one cluster of cases (three nosocomial cases) was detected in the haematology unit. Forty-three percent of cases (N=32) were hospitalized in the haematology unit, and all other cases were hospitalized elsewhere. This three-year survey found a high rate of non-nosocomial IA cases and a high frequency of IA cases hospitalized in units other than haematology. Thus, this study shows the importance of IA surveillance in haematology units and all high-risk units. PMID- 16257085 TI - Implementation of isolation precautions: role of a targeted information flyer. AB - Isolation precautions are advocated in most countries for patients harbouring multi-drug-resistant organisms. We evaluated the impact of a targeted information flyer on the implementation of isolation precautions for patients with methicillin-resistant Staphylococcus aureus (MRSA). A flyer providing a quick reference to the steps to be implemented following detection of MRSA was sent systematically by the microbiologists with all results displaying MRSA. Infection control staff compared isolation precautions in two three-month periods: the first (control) period before implementation of the flyer and the second (intervention) period after implementation of the flyer. Compared with the control period, compliance with isolation precautions increased significantly in the intervention period. In intensive care units, there was a sign posted on the door for 31 of 38 (82%) patients in the control period, and 33 out of 34 (97%) in the intervention period (P=0.06). Use of gowns increased from 82% to 100% (P=0.01), use of dedicated materials increased from 84% to 100% (P=0.03), availability of alcohol hand rub increased from 82% to 94% (P=0.10), and the proportion of MRSA patients in private rooms increased from 71% to 91% of the cases (P=0.07). In conclusion, compliance with isolation precautions increased after attaching a flyer to all MRSA-positive bacteriological results. PMID- 16257086 TI - Effectiveness of protocols for preventing occupational exposure to blood and body fluids in Dutch hospitals. AB - Compliance of different healthcare workers (HCWs) (nurses, physicians, laboratory technicians and cleaners) with protocols to prevent exposure to blood and body fluids (BBF) was studied. Questionnaires were used to assess perception of risks, familiarity with protocols, motivation and actual behaviour. Performance of the protocols in practice was also tested. The practical test provided more reliable results than the questionnaire. HCWs overestimated their knowledge and skills, and compliance was influenced by risk perception. HCWs encountered problems with comprehension, acceptability and applicability of protocols, especially for post exposure precautions. Protocols are not tailored to the differences in knowledge, risk perception and practical needs of different professional groups, probably because HCWs have rarely been involved in writing them and they are governed more by legal considerations than applicability. Most HCWs experienced a lack of organizational support to aid compliance. To improve compliance, we recommend information and training on risk management and individual responsibilities regarding the safety of coworkers and patients, participation of HCWs in protocol development, and support of management to avoid reversion to previous habitual behaviour. PMID- 16257087 TI - Bloodstream infection related to heparin infusion caused by Agrobacterium tumefaciens. PMID- 16257088 TI - Pre-operative antiseptics: the evidence. PMID- 16257089 TI - Quality control of expressed breast milk. PMID- 16257090 TI - Inappropriate inclusion of methicillin-resistant Staphylococcus aureus on death certificates. PMID- 16257091 TI - Risk factors for the acquisition of vancomycin-resistant enterococci during a single-strain outbreak at a major Australian teaching hospital. PMID- 16257093 TI - Coagulase-negative methicillin-resistant Staphylococcus aureus. PMID- 16257092 TI - Economic burden associated with multi-resistant Gram-negative organisms compared with that for methicillin-resistant Staphylococcus aureus in a university teaching hospital. AB - The aim of this study was to investigate the hospital costs of patients with multi-drug-resistant Gram-negative bacilli (MR-GNB) compared with those for patients with methicillin-resistant Staphylococcus aureus (MRSA), using the Austrian performance-related hospital financing system (LKF). The study was performed retrospectively at Vienna General Hospital, a 2,160-bed university teaching hospital, from January to June 2002. There were 99 patients in the MR GNB group (median age 58 years) and 74 patients in the MRSA group (median age 60 years). More patients in the MR-GNB group (59 patients, 60%) were treated in the intensive care unit compared with patients in the MRSA group (25 patients, 34%) (P<0.01). The median hospital stay (42 and 37 days, respectively) and mortality (18 and nine deaths, respectively) of the two groups were similar. The total hospital cost for patients in the MR-GNB group was higher [4 915 712 LKF credit points (median: 34,180) equivalent to 2,605,327 pounds (median: 18,115 pounds )] than that for patients in the MRSA group [2,088,904 LKF credit points (median: 12,650) equivalent to 1, 093, 906 pounds (median: 6,624 pounds)] (P<0.01). This study is limited by being retrospective and having charge-based costings. However, it suggests that the hospital costs of patients with MR-GNB are substantial and may be greater than those of patients with MRSA. PMID- 16257094 TI - Genetic correlates of behavioral endophenotypes in Alzheimer disease: role of COMT, 5-HTTLPR and APOE polymorphisms. AB - Several studies have been conducted to understand the genetic correlates of Alzheimer disease (AD)-related behavioral and psychological symptoms in dementia (BPSD). However, given that BPSD rarely occur in isolation, it has been suggested that targeting BPSD individually is too narrow of an approach if one wants to accurately define all the associated risk factors. To date, we know of no work on genetic polymorphisms related to behavioral endophenotypes in AD. The present study sought to evaluate the relationship between such behavioral endophenotypes in AD and genetic variations in dopamine- or serotonin-related genes, such as catechol-O-methyltransferase (COMT) or 5-HTT gene-linked promoter region (5 HTTLPR), and apolipoprotein E (APOE). Among 232 AD patients who underwent clinical and neuropsychological examination, a behavioral and psychiatric evaluation, and genotyping at COMT, 5-HTTPLR, and APOE; 66.4% showed more than one behavioral symptom. By Principal Component Analysis of Neuropsychiatric Inventory (NPI) symptoms four endophenotypes were identified, these were termed "psychosis", "moods", "apathy", and "frontal". Modeling NPI symptom-endophenotype genotype relationships, and taking into account possible confounds (i.e. demographic characteristics, comorbidities, concomitant pharmacological treatments, and disease severity) by latent variable models, COMT and 5-HTTLPR genetic variations correlated with "frontal" and "psychosis" endophenotypes. APOE genotype did not correlate with any endophenotype. These findings suggest that the possibility of identifying distinct phenotypes on a genetic basis among AD patients exists, and suggest that clustering of BPSD into endophenotypes might provide a new strategy for guiding future research on this issue. PMID- 16257095 TI - Current knowledge of chromosome 12 susceptibility genes for late-onset Alzheimer's disease. AB - In the last decade, it has become more apparent the important role genes play in the development of late-onset Alzheimer's disease (AD). Great efforts, involving human genome scans and candidate gene studies, have been given towards identifying susceptibility genes for AD. A number of regions on different chromosomes have been reported to demonstrate linkage for AD. Of these, findings on chromosome 12 are some of the most compelling. Worldwide genetic association studies pre-dating and subsequent to recent linkage studies have identified and focused upon a number of genes that map to the areas of reported linkage on chromosome 12, however, analyses of those genes studied to date, on the whole, remain inconclusive and ambiguous. This paper reviews studies that have provided evidence of linkage for AD on chromosome 12 and in turn discusses the work conducted to date on candidate genes that have been identified and map to the chromosome 12 regions of interest. PMID- 16257096 TI - Immunization with Neisseria meningitidis outer membrane vesicles prevents bacteremia in neonatal mice. AB - Although vaccines based on outer membrane vesicles (OMV) of Neisseria meningitidis have been developed and administered to children, little is known about the magnitude and quality of the immune response in animal models of early life immunization. We investigated the immunogenicity of meningococcal OMV, and the influence of route and immunization schedule, in neonatal mice. The administration of two intraperitoneal doses of OMV, given at 7 and 14 days after birth, induced a significant antibody response and was highly effective in conferring protection against bacteremia in 21-day-old mice challenged with meningococci. Intranasal immunization was less effective and did not generate a protective immune response. The antibodies elicited by intraperitoneal immunization were cross-reactive with several meningococcal strains and a memory response was demonstrated when mice immunized as neonates were given a booster immunization at 6 weeks of age. PMID- 16257097 TI - Grey variants of the live vaccine strain of Francisella tularensis lack lipopolysaccharide O-antigen, show reduced ability to survive in macrophages and do not induce protective immunity in mice. AB - Francisella tularensis live vaccine strain (LVS) produces two colony types when grown on solid media, often referred to as blue variants (BV) and grey variants (GV). Whereas blue variant bacteria possessed a lipopolysaccharide O-side chain, grey variant bacteria lacked O-side chains. Grey variant bacteria appeared in stationary phase bacterial cultures and could be identified using a novel FACS based assay. Compared to blue variant bacteria, grey variants showed a reduced ability to infect and survive in macrophages. The immunisation of mice with blue variant bacteria, but not grey variant bacteria, induced protective immunity towards fully virulent F. tularensis. PMID- 16257098 TI - Reduced doses of oral killed enterotoxigenic Escherichia coli plus cholera toxin B subunit vaccine is safe and immunogenic in Bangladeshi infants 6-17 months of age: dosing studies in different age groups. AB - The oral-formalin inactivated whole cell enterotoxigenic Escherichia coli (ETEC) vaccine needs to be further tested in developing countries in order to determine the dose at which it will be safe and immunogenic for infants who are the target population for the vaccine. To determine the immunogenicity of reduced doses, studies were first carried out in children, 2-12 years of age (n = 60). The full, half or a quarter doses of the vaccine were comparable in immunogenicity with similar frequency of responses seen to the different antigens (P = NS). Following this result, a pilot study carried out in infants, 6-17 months of age (n = 50), showed that the frequency of episodes of vomiting was lowest when a quarter of the full dose was used. The infants however showed comparable immune responses to the half and quarter dose of vaccine that was tested (P = NS). Based on these results in the infants, a randomized double blind placebo-controlled Phase II study was carried out in 158 children, 6-17 months of age, where a quarter dose of the ETEC vaccine was tested. Adverse events of mild vomiting were seen in only 4% of vaccinees and in 2.5% of placebo recipients. The IgA-antibody secreting cell (ASC) responses to CFA/I (GM: 28.1 ASC/10(7) PBMC) and BS (GM: 55.7 ASC/10(7) PBMC) were elevated compared to placebo recipients (CFA/I-2.0; BS-4.8 ASC/10(7) PBMC) (P = 0.01 to < 0.001). The plasma-IgA antibody titers in vaccinees were also significantly elevated to CFA/I (GM-93.00), CS1 (GM-62.0), CS2 (GM-55.0), CS4 (GM-66.0) and BS (1057.0) compared to preimmune levels or responses or levels in placebo recipients (P < or = 0.05-0.001). This study thus demonstrates that reduced doses of the ETEC vaccine is immunogenic in children and infants as well as safe in infants down to 6 months of age. PMID- 16257099 TI - A novel live recombinant mycobacterial vaccine against bovine tuberculosis more potent than BCG. AB - Mycobacterium bovis infection of cattle and other domesticated animals exacts a significant economic toll in both economically developing and industrialized countries. Vaccination of herds and/or wild animals that share their grazing land and serve as reservoirs of infection has been proposed as a strategy to combat bovine tuberculosis. However, the only currently available vaccine, M. bovis Bacille Calmette-Guerin (BCG), is not highly efficacious. Here we show that a live recombinant vaccine, rBCG30, which expresses large amounts of the Mycobacterium tuberculosis 30 kDa major secretory protein, is more efficacious against bovine tuberculosis than BCG in the highly demanding guinea pig model of pulmonary tuberculosis. Compared with the parental wild-type BCG strain, rBCG30 administered intradermally induced significantly greater cell-mediated and humoral immune responses against the 30 kDa protein, as determined by measuring cutaneous delayed-type hypersensitivity and antibody titers. As for potency, in three independent experiments, rBCG30 induced greater protective immunity than BCG against aerosol challenge with a highly virulent strain of M. bovis, reducing the burden of M. bovis by 0.4 +/- 0.2 log colony-forming units (CFU) in the lung (P < 0.05) and by 1.1 +/- 0.4 log CFU in the spleen (P = 0.0005) below the level in BCG-immunized animals. A recombinant BCG vaccine overexpressing the identical M. bovis 30 kDa protein, rBCG30Mb, also induced greater cell-mediated and humoral immunity against the 30 kDa protein than BCG and greater protective immunity against M. bovis challenge; however, its potency was not significantly different from rBCG30. As rBCG30 is significantly more potent than BCG against M. bovis challenge, it has potential as a vaccine against bovine tuberculosis in domesticated animals and in wild animal reservoirs. PMID- 16257100 TI - Revision surgery after carpal tunnel release--analysis of the pathology in 200 cases during a 2 year period. AB - Carpal tunnel release (CTR) is regarded as a common and successful operative procedure in hand surgery. However, an increasing number of patients with complications have been referred to our hospital. This retrospective investigation was undertaken to clarify the reasons for persisting or recurrent symptoms in 200 patients who underwent secondary exploration during a 26 month period at a single institution. In 108 cases, the flexor retinaculum was found to have been released incompletely. In 12 patients, a nerve laceration had occurred during the primary intervention. In 46 patients, symptoms were due to the nerve being tethered in scar tissue. The re-exploration revealed circumferential fibrosis around and within the median nerve in 17 patients and a tumour in the carpal tunnel in four patients. In 13 patients, no specific reason was found for recurrence of symptoms. We conclude that CTR seems to be a widely underestimated procedure and revision surgery could be largely avoided by reducing technical errors during the primary operation. PMID- 16257101 TI - Aluminum-based drinking-water treatment residuals: a novel sorbent for perchlorate removal. AB - Perchlorate contamination of aquifers and drinking-water supplies has led to stringent regulations in several states to reduce perchlorate concentrations in water at acceptable levels for human consumption. Several perchlorate treatment technologies exist, but there is significant cost associated with their use, and the majority of them are unable to degrade perchlorate to innocuous chloride. We propose the use of a novel sorbent for perchlorate, i.e. an aluminum-based drinking-water treatment residual (Al-WTR), which is a by-product of the drinking water treatment process. Perchlorate sorption isotherms (23+/-1 degrees C) showed that the greatest amount (65%) of perchlorate removed by the Al-WTR was observed with the lowest initial perchlorate load (10 mg L(-1)) after only 2 h of contact time. Increasing the contact time to 24 h, perchlorate removal increased from 65 to 76%. A significant correlation was observed between the amounts of perchlorate removed with evolved chloride in solution, suggesting degradation of perchlorate to chloride. PMID- 16257102 TI - Arsenic speciation, and arsenic and phosphate distribution in arsenic hyperaccumulator Pteris vittata L. and non-hyperaccumulator Pteris ensiformis L. AB - This study examined the roles of arsenic translocation and reduction, and P distribution in arsenic detoxification of Pteris vittata L. (Chinese Brake fern), an arsenic hyperaccumulator and Pteris ensiformis L. (Slender Brake fern), a non arsenic hyperaccumulator. After growing in 20% Hoagland solution containing 0, 133 or 267 microM of sodium arsenate for 1, 5 or 10 d, the plants were separated into fronds, rhizomes, and roots. They were analyzed for biomass, and concentrations of arsenate (AsV), arsenite (AsIII) and phosphorus. Arsenic in the fronds of P. vittata was up to 20 times greater than that of P. ensiformis, yet showing no toxicity symptoms as did in P. ensiformis. While arsenic was concentrated primarily in the fronds of P. vittata as arsenite it was mainly concentrated in the roots of P. ensiformis as arsenate. Arsenic reduction in the plants took longer than 1-d. P. vittata maintained greater P in the roots while P. ensiformis in the fronds. The high arsenic tolerance of the hyperaccumulator P. vittata may be attributed to its ability to effectively reduce arsenate to arsenite in the fronds, translocate arsenic from the roots to fronds, and maintain a greater ratio of P/As in the roots. PMID- 16257103 TI - Attention, short-term memory, and action selection: a unifying theory. AB - Cognitive behaviour requires complex context-dependent processing of information that emerges from the links between attentional perceptual processes, working memory and reward-based evaluation of the performed actions. We describe a computational neuroscience theoretical framework which shows how an attentional state held in a short term memory in the prefrontal cortex can by top-down processing influence ventral and dorsal stream cortical areas using biased competition to account for many aspects of visual attention. We also show how within the prefrontal cortex an attentional bias can influence the mapping of sensory inputs to motor outputs, and thus play an important role in decision making. We also show how the absence of expected rewards can switch an attentional bias signal, and thus rapidly and flexibly alter cognitive performance. This theoretical framework incorporates spiking and synaptic dynamics which enable single neuron responses, fMRI activations, psychophysical results, the effects of pharmacological agents, and the effects of damage to parts of the system to be explicitly simulated and predicted. This computational neuroscience framework provides an approach for integrating different levels of investigation of brain function, and for understanding the relations between them. The models also directly address how bottom-up and top-down processes interact in visual cognition, and show how some apparently serial processes reflect the operation of interacting parallel distributed systems. PMID- 16257104 TI - Successful treatment of persistent vulvodynia with submucous infiltration of betamethasone and lidocaine. PMID- 16257105 TI - The common variant N372H in BRCA2 gene may be associated with idiopathic male infertility with azoospermia or severe oligozoospermia. AB - OBJECTIVE: To explore the possible association between the common single nucleotide polymorphism N372H in human breast cancer susceptibility gene 2 (BRCA2) and the idiopathic male infertility with azoospermia or severe oligozoospermia. STUDY DESIGN: The study included 240 infertile patients with idiopathic azoospermia or severe oligozoospermia and 250 fathered controls. The allele and genotype frequencies of the polymorphism N372H in BRCA2 gene were investigated in both patients and controls using denaturing high performance liquid chromatography analysis (DHPLC). RESULTS: The frequency of allele H of the polymorphism N372H in patients was significantly higher than that of the controls (23.5% versus 17.6%, OR = 1.49, 95% CI 1.06-1.97, P = 0.02) and the subjects bearing rare allele H (NH + HH) significantly increased in patients compared with controls (41.7% versus 32.4%, 95% CI 1.03-2.15, P = 0.03). CONCLUSION: The results of this study suggested that the polymorphism N372H in BRCA2 gene may be associated with idiopathic male infertility with azoospermia or severe oligozoospermia. PMID- 16257107 TI - Androgen receptor and TGFbeta1/Smad signaling are mutually inhibitory in prostate cancer. AB - PURPOSE: The androgen receptor (AR) has been reported to modulate TGFbeta1/Smad signaling and is, like transforming growth factor beta 1 (TGFbeta1) often overexpressed in hormone refractory prostate cancer (HRPC). In human prostate cancer cell lines the role of AR in the response to TGFbeta1 is evaluated. MATERIAL AND METHODS: PC3 cells grow hormone independently, lack AR expression, and have a functioning TGFbeta1/Smad signaling cascade whereas LNCaP cells express (a mutated) AR, and lack TGFbeta receptor (TGFbetaR2) expression. Luciferase reporter assays for AR signaling, TGFbeta1/Smad signaling, and E2F transcriptional activity were performed. PC3 cells and TGFbetaR2 stably transfected LNCaP cells (LNCaP-R2) were incubated with dihydrotestosterone (DHT), or TGFbeta1. Wst-1 assay and flowcytometric evaluation of annexin-V staining were applied to quantify cell growth and apoptosis. Immunoblot analysis was performed to evaluate c-Myc expression. RESULTS: Luciferase reporter assays showed mutual transcriptional inhibition of AR and TGFbeta/Smad signaling in AR transfected PC3 and LNCaP-R2 cells. AR expression reduced the TGFbeta1/Smad transcriptional activity and the growth inhibitory effects of TGFbeta1 also in the absence of DHT in PC3 cells. TGFbeta1 reduced the E2F transcriptional activity of AR activation by DHT. This was associated with a reduced c-Myc expression in PC3 cells. AR expression in PC3 cells prevented TGFbeta1 induced growth inhibition and apoptosis. CONCLUSION: AR overexpression is an effective way of hormone refractory prostate cancer cells to overcome the growth inhibitory effects of elevated serum TGFbeta1 levels even in the absence of DHT. These findings provide an explanation for how AR overexpression favors growth in HRPC. PMID- 16257108 TI - Urinary NMP22 BladderChek test in the diagnosis of superficial bladder cancer. AB - OBJECTIVE: To study the diagnostic efficacy of the NMP22 BladderChek test and to compare it to cytology in the detection of bladder cancer. METHODS: We evaluated 106 voided urinary specimens of patients with suspicion of bladder cancer. All voided urine samples were evaluated by the NMP22 BladderChek test, cytology, sediment and culture. The diagnostic value of the NMP22 BladderChek test was evaluated according to correlation with cystoscopic findings and, in case of tumour, histological findings. A negative test result in a pTaG1 tumour was not considered false-negative in this study. The results were compared to the diagnostic value of cytology. Moreover, the value of the combination of cytology and the NMP22 BladderChek test was determined. RESULTS: In total, 29 patients had histologically proven transitional cell carcinoma of the bladder. The NMP22 BladderChek test detected 40% of 15 pTa tumours and 83.3% of the 6 pT1 tumours. Cytology detected pTa in 33.3% and pT1 in 66.6%. The 1 CIS lesion was detected by cytology. In the group of patients in follow-up the sensitivity and specificity were 57.1% (CI 28.8-82.3) and 89.8% (CI 79.2-96.2) for the NMP22 BladderChek test and 42.9% (CI 17.7-71.7) and 93.2% (CI 83.5-98.1) for cytology. CONCLUSION: The NMP22 BladderChek test has a slightly higher sensitivity compared to cytology, without a relevant loss in specificity. Furthermore it is an easy test with instant result. However, no extra tumours were detected by adjunction of the NMP22 BladderChek test. PMID- 16257109 TI - Risks and benefits of hormonal manipulation as monotherapy or adjuvant treatment in localised prostate cancer. AB - A round table meeting was held to discuss the role of hormonal therapy in localised prostate cancer. The findings of the group were that immediate hormonal therapy does not provide an overall survival advantage in localised and locally advanced prostate cancer. Bicalutamide can prolong disease free survival in patients with locally advanced prostate cancer, however it is important to underline that at this time it has not been shown to influence disease specific nor overall survival. It remains also unproven that early treatment is superior to treatment at progression. However, a trend towards decreased survival with bicalutamide was observed in low risk patients such as those with localised disease. In patients receiving bicalutamide, there were increased cardiovascular side-effects, in addition to the high incidence of gynaecomastia. Early hormonal therapy has to be balanced against such side-effects and the inevitable appearance of hormone refractory disease in patients who progress after hormonal therapy. Consequently, patients with localised, low risk disease are not considered appropriate candidates for hormonal therapy used either as mono therapy or in the adjuvant setting. PMID- 16257110 TI - Re: Wespes E, Rammal A, Christian G. Sildenafil non-responders: haemodynamic and morphometric studies. Eur urol 2005;48:136-9. PMID- 16257111 TI - Improving the preservation of the urethral sphincter and neurovascular bundles during open radical retropubic prostatectomy. AB - OBJECTIVES: To describe a technique for open nerve-sparing radical retropubic prostatectomy. METHODS: The technique basically implies incising the levator and prostatic fasciae high anteriorly (1 and 11 o'clock positions) over the prostate, developing the plane between the prostatic capsule and prostatic fascia, and displacing the neurovascular network localized between the two fasciae laterally. This allows for a minimal-touch dissection of the external urethral sphincter and a very efficient dissection of the neurovascular bundles at the level of membranous urethra and prostatic apex. RESULTS: Forty-two patients underwent a bilateral nerve-sparing operation and were followed- up for 6 months. Six patients (14.3%) had positive margins: 4 patients had pT2 disease (in all, the positive margin was monofocal) and 2 patients had pT3 disease (both had multifocal positive margins). Continence (defined as being dry or having one pad remain dry for 24 hours) was achieved in 44% of patients at catheter removal, and in 60%, 72%, and 90% of patients at the 1-, 3-, and 6-month follow-up visits. Potency (defined as an erectile function domain score > or =26) was obtained in 15%, 40%, and 52% of patients at the 1-, 3-, and 6-month follow-up visits. All patients used a PDE5-inhibitor during the investigation period. CONCLUSIONS: These preliminary results suggest that the high incision of the levator and prostatic fasciae may facilitate efficient preservation of the external urethral sphincter and the neurovascular bundles innervating the corpora cavernosa and the sphincter. PMID- 16257113 TI - An unusual spinal arachnoiditis. AB - Cryptococcal spinal arachnoiditis occurs in patients with meningitis and usually when they are immunocompromised. Spinal symptoms in cryptococcosis are rare and a very exceptional entity in the immunocompetent population. We present a young immunocompetent male who developed progressively increasing paraparesis due to primary cryptococcal arachnoiditis, who showed significant improvement after antifungal therapy. Although extremely rare, spinal arachnoiditis in an immunocompetent individual can be caused due to cryptococcus, as in our case. This case illustrates and emphasizes the necessity for an exhaustive and complete investigation, with a high index of suspicion for fungal etiology in patients presenting with spinal arachnoiditis or other disabling, progressive spinal cord syndromes of unknown etiology. Awareness of this presentation is necessary to avoid delay in diagnosis and management of this potentially curable condition. PMID- 16257112 TI - Progressive multifocal leukoencephalopathy in an HTLV-I carrier. AB - This report describes a previously 28-year-old healthy woman, identified as an asymptomatic human T-lymphotropic virus type I (HTLV-I) carrier, who developed both progressive multifocal leukoencephalopathy (PML) and Pneumocystis jiroveci pneumonia. For diagnostic confirmation of PML, stereotactic brain biopsy demonstrated multiple demyelinating lesions with the presence of JC viral antigen. Intramuscular alpha-interferon therapy for 2 weeks brought considerable neurologic improvement. Three years later, the patient developed lymphoma-type of adult T-cell leukemia, suggesting that HTLV-I carrier might be one of the underlying diseases of PML. PMID- 16257114 TI - COD, para-chlorophenol and toxicity removal from para-chlorophenol containing synthetic wastewater in an activated sludge unit. AB - Chlorinated phenolic compounds present in some chemical industry wastewaters cause severe toxic effects on the organisms and often are resistant to biological degradation. Synthetic wastewater containing different concentrations of para chlorophenol (4-chlorophenol, 4-CP) was biologically treated in an activated sludge unit for COD, 4-CP and toxicity removal. Effects of feed 4-CP concentration on COD, 4-CP, toxicity removals and on sludge volume index were investigated at a constant sludge age of 20 days and hydraulic residence time (HRT) of 25 h. Resazurin method based on dehydrogenase activity was used for determination of the toxicity of the feed and effluent wastewater. COD and 4-CP removals were not affected by the presence of 4-CP in the wastewater up to feed 4 CP concentration of 925 mg l(-1) because of almost complete degradation of 4-CP yielding lower than 50 mg l(-1) 4-CP in the aeration tank. Percent COD, 4-CP and toxicity removals decreased and the effluent COD, 4-CP and toxicity levels increased with further increases in the feed 4-CP concentrations above 925 mg l( 1) because of inhibitory concentrations of 4-CP in the reactor. Biomass concentration in the aeration tank decreased and the sludge volume index (SVI) increased with feed 4-CP concentrations above 925 mg l(-1) resulting in lower COD and 4-CP removal rates. The rates of COD and 4-CP removals indicated substrate (4 CP) inhibition for the feed 4-CP concentrations above 925 mg l(-1) corresponding to the reactor 4-CP of above 200 mg l(-1). The system should be operated at the feed 4-CP concentrations of less than 900 mg l(-1) (4-CP(R) < 200 mg l(-1)) in order to obtain high rates and extents of COD, 4-CP and toxicity removals at a sludge age of 20 days and HRT of 25 h. PMID- 16257115 TI - Adsorption equilibrium of phenol onto chemically modified activated carbon F400. AB - In this work the adsorption of phenol solutions onto activated carbon Calgon F400 has been studied. The carbon was modified by acid treatment, using soxhlet extraction with hydrochloric acid 2N for 120 cycles. The treatment did not affect significantly the surface area of the activated carbon but affected significatively the different functional groups, and thus its adsorption properties. Toth model reproduced satisfactorily the experimental isotherm data and an adsorption enthalpy of -17.9 kJ mol-1 was found, which indicates that the process is exothermic. The pH affected significatively to the adsorption process and an empirical polynomial equation was able to reproduce maximum capacity as a function of pH. The isotherms obtained at pH 3 and 7 are very similar and showed a higher adsorption capacity compared with that obtained at pH 13. The use of phosphate buffer solutions decreased the maximum phenol adsorption capacity, due to the competitive adsorption between the phenol and phosphates. Finally, it was demonstrated that the acid treatment introduced chloride ions into the carbon, giving it properties of ion exchanger. PMID- 16257117 TI - The adsorption kinetics of the cationic dye, methylene blue, onto clay. AB - In this study, adsorption kinetics of a cationic dye, methylene blue, onto clay from aqueous solution with respect to the initial dye concentration, temperature, pH, mixing rate and sorbent dosage were investigated. In order to understand the adsorption mechanism in detail, zeta potentials and the conductivities of clay suspensions at various pH (1-11) and cation exchange capacity (CEC) were measured. Porosity and BET surface area of clay studied were determined. The results showed that the adsorption has been reached to equilibrium in 1 h. It was found that the amount adsorbed of methylene blue increases with decreasing temperature and also with increasing both sorbent dosage and increasing initial dye concentration. Adsorption capacity decreases with increasing pH, except for the natural pH (5.6) of clay suspensions. The adsorption kinetics of methylene blue has been studied in terms of pseudo-first-order, pseudo-second-order sorption and intraparticle diffusion processes thus comparing chemical sorption and diffusion sorption processes. It was found that the pseudo-second-order mechanism is predominant and the overall rate of the dye adsorption process appears to be controlled by the more than one-step. PMID- 16257116 TI - TiO2/UV/O3-BAC processes for removing refractory and hazardous pollutants in raw water. AB - TiO2/UV/O3-BAC (biological activated carbon) process was employed to treat raw water and compared to UV/O3-BAC process in its optimum parameters (3 mg/L ozone dosage with 15 min oxidation time and 15 min empty bed contact time in BAC). The results showed that the presence of TiO2 improved ozone utilization and biodegradability of the effluent. For the dissolved organic carbon (DOC) removal, TiO2/UV/O3-BAC was more efficient than UV/O3-BAC and its synergetic effect is more than that in UV/O(3)-BAC process. It was showed that small molecules with MW<3000 Da predominated in the raw water accounting for more than 56% DOC, they were increased after oxidation, accounting for more than 64% DOC. GC/MS analysis showed that TiO2/UV/O3-BAC process was effective in removing phthalate esters (PAEs) and persistent organic pollutants (POPs). PAEs' removal ratio reached more than 94% and reduced with the increase of the length of the alkyl side chains and the alkyl branch chains. TiO2/UV/O3-BAC process was also very effective in removing POPs. Polybromobiphenyls' removal rate reached more than 89% and decreased with the increase of substitutional bromines except for 2,2',5,5' tetrabromobiphenyl, which can be completely removed. PMID- 16257118 TI - SELPLG and SELP single-nucleotide polymorphisms in multiple sclerosis. AB - P-Selectin (SELP) and P-selectin glycoprotein ligand-1 (SELPLG) constitute a receptor/ligand complex involved in the recruitment of activated lymphocytes, a critical event in the pathogenesis of multiple sclerosis (MS). In order to determine whether genetic variation in these pivotal molecules influences susceptibility to MS, we genotyped 214 Italian patients compared with 220 Italian controls for three single-nucleotide polymorphisms (SNPs): SELPLG Met62Ile, SELP C-2123G and SELP Thr715Pro. No significant differences in both SELP SNPs were found between patients and controls, whereas a decreased frequency of the Met62Ile SNP was found in patients versus controls in the Italian population (P = 0.025). To confirm these preliminary findings, the Met62Ile SNP was analysed in 938 UK trio families. This SNP did not show evidence for association with susceptibility to MS in the larger UK cohort. Therefore, none of the SNPs investigated is associated with MS, although this analysis does not conclusively exclude SELPLG and SELP as genetic risk factors for MS as much variation remains untested. PMID- 16257119 TI - Attention modulates the processing of emotional expression triggered by foveal faces. AB - To investigate whether the processing of emotional expression for faces presented within foveal vision is modulated by spatial attention, event-related potentials (ERPs) were recorded in response to stimulus arrays containing one fearful or neutral face at fixation, which was flanked by a pair of peripheral bilateral lines. When attention was focused on the central face, an enhanced positivity was elicited by fearful as compared to neutral faces. This effect started at 160 ms post-stimulus, and remained present for the remainder of the 700 ms analysis interval. When attention was directed away from the face towards the line pair, the initial phase of this emotional positivity remained present, but emotional expression effects beyond 220 ms post-stimulus were completely eliminated. These results demonstrate that when faces are presented foveally, the initial rapid stage of emotional expression processing is unaffected by attention. In contrast, attentional task instructions are effective in inhibiting later, more controlled stages of expression analysis. PMID- 16257120 TI - Tubulin nitration in human gliomas. AB - Immunohistochemical and biochemical investigations showed that significant protein nitration occurs in human gliomas, especially in grade IV glioblastomas at the level of astrocytes and oligodendrocytes and neurones. Enhanced alpha tubulin immunoreactivity was co-present in the same elements in the glioblastomas. Proteomic methodologies were employed to identify a nitrated protein band at 55 kDa as alpha-tubulin. Peptide mass fingerprinting procedures demonstrated that tubulin is nitrated at Tyr224 in grade IV tumour samples but is unmodified in grade I samples and in non-cancerous brain tissue. These results provide the first characterisation of endogenously nitrated tubulin from human tumour samples. PMID- 16257122 TI - Hyperglycemia increases superoxide production in the CA1 pyramidal neurons after global cerebral ischemia. AB - Transient global cerebral ischemia results in selective neuronal death in the vulnerable hippocampal CA1 pyramidal neurons in a delayed manner. Hyperglycemia accelerates and exacerbates neuronal damage in this region. The object of this study was to determine whether hyperglycemia-enhanced damage is associated with increased production of superoxide anion after ischemia. The results showed that hyperglycemic ischemia caused a significant increase of superoxide production in the hippocampal CA1 neurons compared to normoglycemic animals after 18 h of recirculation, suggesting that enhanced superoxide anion production may mediate the hyperglycemia-accelerated and -enhanced neuronal death in the hippocampal CA1 area after ischemia and reperfusion. PMID- 16257123 TI - Analysis of the PINK1 gene in a cohort of patients with sporadic early-onset parkinsonism in Taiwan. AB - Mutations in the PINK1 gene have been shown to cause autosomal recessive Parkinson's disease (PD) and/or early onset sporadic PD in Italy, Spain, North America, Ireland, and Asia. However, there are limited data on PINK1 mutations in sporadic early onset Asian PD patients. To determine the prevalence of PINK1 mutation in Taiwanese population, we conducted genetic analysis of PINK1 mutation in 73 early onset sporadic PD and 94 normal control subjects. We only identified a novel single heterozygous mutation R 407Q mutation in exon 6 of this gene in one patient at the age onset of 54. Overall, these data indicate that PINK1 mutations are rare in our population. Based on our results, unless common mutational hotspots are identified, routine testing for this mutation at least in our population may not be cost-effective. PMID- 16257121 TI - Apoptotic effect of ethanol is potentiated by caffeine-induced calcium release in rat astrocytes. AB - In this study, we investigated agents that increased intracellular calcium levels and their correlation with apoptotic cell death induction. We used rat astrocytes to investigate the increase in cytosolic Ca2+ (Ca(c)2+) and apoptosis induction by drugs that mobilize Ca2+ from different sources. We observed that thapsigargin (Thap), caffeine (Caff) and FCCP which caused similar increases in Ca(c)2+ levels (30-40%), also induced similar apoptotic rates (30-35%). On the other hand, antimycin (Anti), staurosporine (STS) and ethanol (Eth) promoted higher increases in Ca(c)2+ (55-65 %) and higher apoptotic rates (55-85%). Eth induced cell death in a concentration- and time-dependent manner. After treatment with Eth plus Caff for 6, 12 and 24 h, these effects were strongly potentiated. Results suggest that there might be a correlation between Ca(c)2+ increase and the rate of apoptosis. It is possible that Eth induces cell death by activation of more than one pathway and Ca2+ might be one of the elements involved. The present work indicates that Ca2+ can potentiate death by ethanol in rat astrocytes. PMID- 16257124 TI - Intrinsic and extrinsic factors of turning preferences in humans. AB - Turning behaviour in 107 adolescents was observed during walking and running under different temporal and biomechanical constraints. Participants ran and walked back and forth between two lines 9.5m apart in a neutral environment. All of the turns that the participants made to change direction between the lines were videotaped. A general preference for turning leftwards was observed with the percentage being higher in the unconstrained running condition when compared to the walking condition (71% versus 59%, respectively). This intrinsic preference was easily overruled when positional constraints on the starting position were imposed. Such positional constraints did not, however, suppress the intrinsic directional bias observed during running. It is concluded that turning preference in humans is the result of a complex interaction between intrinsic preferences and externally imposed task constraints. PMID- 16257125 TI - Preventing radiation induced xerostomia. AB - Radiation induced xerostomia is a frequent consequence of radiotherapy (RT) for head-neck cancer (HNC) patients, when parotid glands are included in the radiation fields. Although early appearing xerostomia may be alleviated with the use of pilocarpine, persistent chronic xerostomia affects more than 70% of HNC patients treated with post-operative or radical radiotherapy and significantly impairs the quality of life potentially cured patients. The present manuscript reviews and discusses the current technological (conformal and intensity modulated RT) and pharmacological (amifostine) developments aiming to prevent the severity and reduce incidence of both acute and late radiation xerostomia in patients with HNC. PMID- 16257126 TI - Cystic neoplasms of the pancreas; what a clinician should know. AB - Primary cystic neoplasms of the pancreas (serous cystic neoplasms, mucinous serous neoplasms, and intraductal papillary mucinous neoplasms) are lesions of emerging importance. With the wide availability of modern imaging methods, these neoplasms are being recognized with increasing frequency. Due to the improvement of these sophisticated imaging techniques, it is often possible to differentiate preoperatively these primary pancreatic cystic neoplasms not only from other cystic pancreatic disorders (such as pancreatic pseudocysts) but also from one another. This differentiation is very important for the clinician, since these neoplasms have radically different biologic behavior. Serous cystic neoplasms are uniformly benign and usually do not mandate resection unless the lesion is symptomatic. In contrast, mucinous cystic neoplasms and intraductal papillary mucinous neoplasms have a premalignant or overtly malignant tendency, and therefore should be managed aggressively by pancreatic resection. In these mucinous cystic neoplasms, recognition of an underlying malignancy is often not possible without a detailed histopathologic examination of the entire resected specimen. In the absence of invasive disease, prognosis is excellent after appropriate surgery. The presence of invasive malignancy signifies a poor prognosis. PMID- 16257127 TI - School-based smoking cessation programs: do youth smokers want to participate in these programs? AB - The purpose of the present study was to examine characteristics that predict interest in school-based cessation programs among 3136 youth smokers with intentions to quit smoking. The majority of youth smokers report that they would not join a school-based smoking cessation program. However, improving awareness of these types of programs among students is important as sub-populations of youth smokers were more likely to be interested in school-based cessation initiatives when aware that such programs exist. Future school-based cessation intervention outcomes might be improved if programs are targeted to the youth that are most likely to use them, if more youth can be made aware of existing programs, and if the benefits of participating in such programs can be more adequately conveyed to youth smokers. PMID- 16257128 TI - The importance of early physical trauma in the development of autism. PMID- 16257129 TI - ROC curves and evaluation of radiation-induced pulmonary toxicity in breast cancer. AB - PURPOSE: To study clinical, radiologic, and physiologic pulmonary toxicity in 128 women after adjuvant radiotherapy (RT) for breast cancer in relation to dosimetric factors. METHODS AND MATERIAL: The patients underwent pulmonary function testing before and 5 months post-RT. Similarly, computer tomography of the chest was repeated 4 months post-RT and changes were scored with a semiquantitative system. Clinical symptoms were registered and scored according to Common Toxicity Criteria. All patients underwent three-dimensional dose planning, and the ipsilateral lung volume receiving > or = 13 Gy (V13), V20, and V30 were calculated. Multiple logistic or regression analyses were used for multivariate modeling. The relation between the dosimetric factors and side effects was also analyzed with receiver operating characteristic (ROC) curves. RESULTS: V20 was, according to multivariate modeling, the most important variable for the occurrence of the three studied side effects (p < 0.01). Age was also related to symptomatic and radiologic pneumonitis. Reduced pre-RT functional level was more common in patients developing symptomatic toxicity. The ROC areas for symptomatic pneumonitis in relation to V13, V20, and V30 were 0.69, 0.69, and 0.67, and for radiologic pneumonitis 0.85, 0.85, and 0.81. CONCLUSIONS: Our results support the use of three-dimensional planning aimed at minimizing the percent of incidentally irradiated lung volume to reduce pulmonary toxicity. Age was also correlated with post-RT side effects. According to ROC analysis, V20 could well predict the risk for radiologic pneumonitis for the studied semiquantitative model. PMID- 16257130 TI - Analysis of gene-expression profiles after gamma irradiation of normal human fibroblasts. AB - PURPOSE: To understand comprehensive transcriptional profile of normal human fibroblast in response to irradiation. METHODS AND MATERIALS: To identify genes whose expression is influenced by gamma radiation, we used a cDNA microarray to analyze expression of 23,000 genes in normal human fibroblasts at 7 timepoints (1, 3, 6, 12, 24, 48, and 72 hours) after 5 different doses (0.5, 2, 5, 15, and 50 Gy) of exposure. RESULTS: Among the genes that showed altered expression patterns, some were already known to be regulated by irradiation, for instance ODC, EGR1, FGF2, PCNA, PKC, and several p53-target genes, including p53DINP1, BTG2, GADD45, and MDM2. The time course of each dose showed that from 350 to 600 genes were affected as to their expression; induction profiles characteristic to each dose were demonstrated. Of the total identified, only 89 genes were up regulated; the vast majority was down-regulated over the 72-hour time course. We identified 21 genes that were distinctly induced by irradiation; 11 of them were functionally known, and 6 of those were p53-target genes. CONCLUSIONS: The results underscored the complexity of the transcriptional responses to irradiation, and the data should serve as a basis for global characterization of radiation-regulated genes and pathways. PMID- 16257131 TI - Fractionated proton beam irradiation of pituitary adenomas. AB - PURPOSE: Various radiation techniques and modalities have been used to treat pituitary adenomas. This report details our experience with proton treatment of these tumors. METHODS AND MATERIALS: Forty-seven patients with pituitary adenomas treated with protons, who had at least 6 months of follow-up, were included in this analysis. Forty-two patients underwent a prior surgical resection; 5 were treated with primary radiation. Approximately half the tumors were functional. The median dose was 54 cobalt-gray equivalent. RESULTS: Tumor stabilization occurred in all 41 patients available for follow-up imaging; 10 patients had no residual tumor, and 3 had greater than 50% reduction in tumor size. Seventeen patients with functional adenomas had normalized or decreased hormone levels; progression occurred in 3 patients. Six patients have died; 2 deaths were attributed to functional progression. Complications included temporal lobe necrosis in 1 patient, new significant visual deficits in 3 patients, and incident hypopituitarism in 11 patients. CONCLUSION: Fractionated conformal proton-beam irradiation achieved effective radiologic, endocrinological, and symptomatic control of pituitary adenomas. Significant morbidity was uncommon, with the exception of postradiation hypopituitarism, which we attribute in part to concomitant risk factors for hypopituitarism present in our patient population. PMID- 16257132 TI - Combination of external-beam radiotherapy with intraoperative electron-beam therapy is effective in incompletely resected pediatric malignancies. AB - PURPOSE: Intraoperative electron-beam radiotherapy (IOERT) has been applied for local dose escalation in over 1,400 patients in Heidelberg since 1991. Among these were 30 children, in 18 of whom IOERT was employed in radiation treatment with external-beam radiotherapy (EBRT) on account of incomplete resection. We address the question whether IOERT is able to compensate for microscopic or macroscopic tumor residue if employed in the overall radiation regimen. METHODS AND MATERIALS: The data of the aforementioned 18 children were analyzed with regard to local recurrence, overall survival, and complication rates. All children suffered from either sarcomas or neuroblastomas. In all children, IOERT was employed for local dose escalation after or before EBRT. RESULTS: After a median follow-up of 60.5 months, 15 of the treated children are alive. One local failure has been observed. Six children show clinically significant late morbidity, including the loss of a treated limb (Radiation Therapy Oncology Group Grade 4 [RTOG 4]), a severe nerve lesion (RTOG 3), an orthopedic complication (RTOG 2), a ureteral stenosis (not clinically significant), and a kidney hypotrophy (not clinically significant). In 1 child a fracture due to radionecrosis (RTOG 4) was diagnosed; however, in the follow-up, local tumor relapse was diagnosed as another possible reason for the fracture. CONCLUSIONS: Regarding the low incidence of local failure, IOERT seems to be able to compensate incomplete tumor resection in childhood sarcoma and neuroblastoma patients. The incidence of late morbidity is low enough to justify the employment of IOERT as part of the radiation treatment regimen for pediatric patients. PMID- 16257133 TI - Quantitative molecular diagnosis of axillary drainage fluid for prediction of locoregional failure in patients with one to three positive axillary nodes after mastectomy without adjuvant radiotherapy. AB - PURPOSE: A quantitative multiple-marker reverse transcriptase (RT)-polymerase chain reaction (PCR) assay for sensitive detection of cancer cells in axillary drainage fluid was developed to examine whether the presence of cancer cells in axillary drainage fluid can be used as a predictor of locoregional recurrence (LRR) in patients with breast cancer who had T1/2 primary tumors and one to three positive axillary lymph nodes treated with modified radical mastectomy without adjuvant radiotherapy. METHODS AND MATERIALS: Axillary drainage fluid was collected from 126 patients with invasive ductal carcinoma of the breast who were treated with modified radical mastectomy and were found to have one to three positive axillary nodes. Cancer cells in axillary drainage fluid were detected by RT-PCR assay using primers specific for carcinoembryonic antigen (CEA) and cytokeratin-19 (CK-19) together with numerous clinicopathologic and treatment related factors and were analyzed for their impact on LRR. RESULTS: A total of 38 patients suffered LRR during follow-up and the multimarker RT-PCR assays for CEA and CK-19 in the axillary drainage fluid both were positive in 34 patients (27.0%), of which 29 patients had LRR. In univariate analysis, the 5-year LRR free survival showed higher rates in patients with PCR-negative findings in axillary drainage fluid (p<0.0001), age>or=40 years old (p<0.0001), tumor size<2.5 cm (p<0.0001), negative lymph-vascular space invasion (p=0.026), and T1 status (<0.0001); in multivariate analysis, PCR-positive findings together with age and tumor size were found to be independent predictors of LRR (all p<0.05). CONCLUSION: Multiplex RT-PCR assay for CEA and CK-19 was highly sensitive for detection and might be useful for prediction of LRR in such subgroup breast cancer patients. PMID- 16257134 TI - Gamma knife radiosurgery for cerebral arteriovenous malformations in children/adolescents and adults. Part I: Differences in epidemiologic, morphologic, and clinical characteristics, permanent complications, and bleeding in the latency period. AB - PURPOSE: To compare the epidemiologic, morphologic, and clinical characteristics of 92 children/adolescents (Group A) and 362 adults (Group B) with cerebral arteriovenous malformations (cAVMs) considered suitable for radiosurgery; to correlate radiosurgery-related permanent complication and post-radiosurgery bleeding rates in the 75 children/adolescents and 297 adults available for follow up. METHODS AND MATERIALS: Radiosurgery was performed with a model C 201-source Co60 Leksell Gamma Unit (Elekta Instruments, Stockholm, Sweden). Fisher exact two tailed, Wilcoxon rank-sum, and two-sample binomial exact tests were used for statistical analysis. RESULTS: There were significant differences between the two populations in sex (p = 0.015), clinical presentation (p = 0.001), and location (p = 0.008). The permanent complication rate was lower in younger (1.3%) than in older patients (5.4%), although the difference was not significant (p = 0.213). The postradiosurgery bleeding rate was lower in Group A (1.3%) than in Group B (2.7%) (p = 0.694), with global actuarial bleeding rates of 0.56% per year and 1.15% per year, respectively. CONCLUSIONS: The different characteristics of child/adolescent and adult cAVMs suggest that they should be considered two distinct vascular disorders. The similar rates of radiosurgery-related complications and latency period bleeding in the two populations show that gamma knife radiosurgery does not expose young patients to a higher risk of sequelae than that for older patients. PMID- 16257135 TI - Extrahepatic portal vein occlusion without recurrence after pancreaticoduodenectomy and intraoperative radiation therapy. AB - PURPOSE: Although there are no definitive studies that characterize the survival benefit of intraoperative radiation therapy (IORT), the therapy does not seem to produce significant complication. In our institution, pancreaticoduodenectomy (PD) and IORT are often complicated by the development of extrahepatic portal vein occlusion (EHPO). The aim of this study was to characterize the phenomenon of EHPO after PD and IORT. METHODS AND MATERIALS: Between September 1992 and December 2001, 107 patients received macroscopic curative PD for periampullary disease in our institution. IORT (radiation dose: 20 Gy) was performed in 53 of these patients. Criteria for diagnosis of EHPO were as follows: (1) computerized tomography findings of occlusive extrahepatic portal vein, (2) symptoms of portal hypertension, and (3) confirmation to exclude tumor recurrence from origin of EHPO, because this study examined whether EHPO was a complication of PD and IORT. RESULTS: EHPO was diagnosed in 12 patients. Among patient and operative variables, IORT was the only statistically significant factor associated with a diagnosis of EHPO (p = 0.0052). The median developed time to EHPO and overall survival after surgery in EHPO patients were 358 days and 2,562 days, respectively. Eight patients (67%) with EHPO died during the follow-up period. At 5 years after therapy, EHPO was diagnosed in 67% of survivors who had received IORT. CONCLUSIONS: Patients undergoing IORT and PD have a relatively high incidence of EHPO, and patients who develop postoperative EHPO have poor prognoses. PMID- 16257136 TI - Effect of aqueous Euphorbia hirta leaf extract on gastrointestinal motility. AB - The aqueous leaf extract of Euphorbia hirta decreased the gastrointestinal motility in normal rats and decreased the effect of castor oil-induced diarrhoea in mice. PMID- 16257137 TI - Multi-level analysis of causal attribution of injury to alcohol and modifying effects: Data from two international emergency room projects. AB - Although alcohol consumption and injury has received a great deal of attention in the literature, less is known about patient's causal attribution of the injury event to their drinking or factors which modify attribution. Hierarchical linear modeling is used to analyze the relationships of the volume of alcohol consumed prior to injury and feeling drunk at the time of the event with causal attribution, as well as the association of aggregate individual-level and socio cultural variables on these relationships. Data analyzed are from 1955 ER patients who reported drinking prior to injury included in 35 ERs from 24 studies covering 15 countries from the combined Emergency Room Collaborative Alcohol Analysis Project (ERCAAP) and the WHO Collaborative Study on Alcohol and Injuries. Half of those patients drinking prior to injury attributed a causal association of their injury with alcohol consumption, but the rate of causal attribution varied significantly across studies. When controlling for gender and age, the volume of alcohol consumed and feeling drunk (controlling for volume) were both significantly predictive of attribution and this did not vary across studies. Those who drink at least weekly were less likely to attribute causality at a low volume level, but more likely at high volume levels than less frequent drinkers. Attribution of causality was also less likely at low volume levels in those societies with low detrimental drinking patterns, but more likely at high volume levels or when feeling drunk compared to societies with high detrimental drinking patterns. These findings have important implications for brief intervention in the ER if motivation to change drinking behavior is greater among those attributing a causal association of their drinking with injury. PMID- 16257138 TI - Prospective multicenter observational study of 260 infants born to 259 opiate dependent mothers on methadone or high-dose buprenophine substitution. AB - Specialized prenatal care and substitution programs improve the perinatal prognoses of pregnant drug-abusers and their infants. Although methadone is well documented, little is known about high-dose buprenorphine (HDB). This prospective, multicenter (n = 35) observational study included 259 women on maintenance during pregnancy: 39% on methadone and 61% on HDB. Major findings were: 46% of them received good prenatal care; 62% had peridural analgesia; 12.3% delivered prematurely (<37 weeks); mean gestational age, 38.6 weeks; mean birth weight, 2822g. Three-quarters of the newborns developed neonatal abstinence syndrome (NAS) beginning at a mean age of 40h, with the mean maximum Lipsitz score of 9.1 at 72 h; half of them were treated, mainly with morphine hydrochloride. No baby died. Newborns were discharged with their mothers (96%) or placed in foster care (4%). Comparing methadone with HDB, respectively, mean age at the maximum Lipsitz score was 81 h versus 66 h (P = 0.066). The perinatal medical and social prognoses for these 259 drug addicts and their infants appeared to be improved by specialized prenatal care and was similar for methadone or BHD substitution during pregnancy. PMID- 16257139 TI - Evaluation of a depression-related model of alcohol problems in 430 probands from the San Diego prospective study. AB - BACKGROUND: Some individuals might use alcohol to help cope with stress and feelings of depression. Studies of such depression or affect-related models have produced divergent results, which might reflect different definitions of affective disturbances, varying emphasis on excluding substance-induced depressions, and different approaches to defining family histories (FH). This paper describes a prospective study that evaluates a model of the role of depressive episodes in the development of alcohol-related problems, while controlling for substance-induced mood disorders. METHODS: Personal interviews were obtained 10 and 15 years after 430 Caucasian subjects entered a study of drinking but not alcohol-dependent 20-year olds. An AMOS Structural Equation Model (SEM) was used to evaluate the development of alcohol problems in the context of the FH of alcoholism and independent (i.e., not substance induced) depressive episodes, personal histories of independent depression, levels of stress, social support, drinking to cope with stress, expectations of the effects of alcohol, and peer drinking. RESULTS: The SEM explained 51% of the variance for the 15-year outcome, and demonstrated good fitness characteristics. The FH of mood disorders (FHdep) predicted depressive episodes through interactions with higher stress and an FH of alcoholism (FHalc), as well as in the context of lower social support. Depressive episodes contributed to the development of alcohol problems both directly and through drinking to cope. CONCLUSIONS: In this model when independent depressive syndromes developed in individuals with a FH of alcoholism, they modestly enhanced the risk for alcohol-related problems, but FHalc did not by itself increase the risk for independent depressions. PMID- 16257140 TI - Denaturing gradient gel electrophoresis gel expansion (DGGEGE)--an attempt to resolve the limitations of co-migration in the DGGE of complex polymicrobial communities. AB - Recent molecular approaches for the study of microbial communities such as PCR cloning have enabled the detection and identification of as-yet-unculturable taxa. Cloning and sequencing of multiple samples is extremely laborious and expensive to perform thoroughly due to the large diversity involved. For this purpose, techniques such as denaturing gradient gel electrophoresis (DGGE) may be better suited. There is increasing evidence suggesting that DGGE of complex polymicrobial communities may be limited by co-migration of different sequences. In this study, we attempt to address this limitation by excising individual bands and running them through a shorter denaturant gradient, a process we have termed "denaturing gradient gel electrophoresis gel expansion" (DGGEGE). PMID- 16257141 TI - Molecular epidemiological characteristics of virulence factors on enteroaggregative E. coli. AB - Escherichia coli with various types of adherence patterns to cultured epithelial cells have been described over the years as being associated with both acute and persistent diarrhea. Most enteroaggregative E. coli (EAEC) strains harbor a 60- to 65-MDa plasmid called pAA which has been shown to encode the aggregative adherence fimbriae AAF/I and AAF/II; the enterotoxin EAST1 and Pet, a serine protease which has been described as causing enterotoxic and cytotoxic effects. Another serine protease denominated Pic, encoded by a chromosomal gene displaying mucinolytic activity, serum resistance, and hemagglutination, has also been associated with EAEC strains. In this study, EAEC strains that isolated from the rectal swab of neonates at the neonatal intensive care unit of Pusan National University Hospital in 2003 were tested for the presence of the pAA using polymerase chain reaction (PCR) and DNA colony hybridization methods. To further characterize these EAEC strains, we used PCR to detect genes for proposed EAEC virulence factors and examined HeLa cell adherence assay, antimicrobial susceptibility test, serotyping, cytotoxicity test and epidemiological characteristics. EAEC isolates found were genotyped by random amplified polymorphic DNA and pulsed-field gel electrophoresis. PMID- 16257142 TI - Murine implantation serine proteinases 1 and 2: structure, function and evolution. AB - Implantation is a vital phase in pregnancy whereupon the hatched embryo invades into the uterine wall to establish intimate contacts with the mother for further development. Although it is generally believed that proteinases are major factors that confer the embryo its invasive character, the nature of proteinases involved in implantation remain mostly elusive. In this article, we review the organization, structure and postulated function of the implantation serine proteinase (ISP1 and 2) genes. The ISPs are embedded within a cluster of tryptase genes on mouse chromosome 17. They are most closely related to members of the mast cell tryptase family, indicating that they may possess some properties characteristic of tryptases including multimerization-dependent activation. The significant similarities found in regulatory regions of ISP genes, together with the observation that ISP proteins are co-expressed and heterodimerize in the embryo and uterus suggests that they are intimately co-regulated during implantation. Inhibition of ISP proteolytic function has implicated this enzyme in the processes of embryo hatching and implantation. PMID- 16257143 TI - Development of real-time diagnostic assays specific for Mycoplasma mycoides subspecies mycoides Small Colony. AB - Rapid and specific detection of Mycoplasma mycoides subsp. mycoides Small Colony (M. mycoides SC) is important for the effective control of contagious bovine pleuropneumonia. Although the United States has been free of this disease for over 100 years, it is necessary to develop modern diagnostic assays that are sensitive and specific for biological agents that would affect the US agricultural industry following accidental or intentional introduction into the US agricultural population. With this aim in mind, we have identified M. mycoides SC-specific genetic loci and developed TaqMan-based PCR assays for the detection of M. mycoides SC. The TaqMan assay allows for real-time detection of specific, amplified PCR products using portable equipment, enabling testing to be performed in the field. These assays are specific for M. mycoides SC, failing to amplify DNA from other organisms belonging to the M. mycoides cluster or two phylogenetically unrelated bovine mycoplasma species. Standard curves were drawn based on the linear relationships measured between the threshold fluorescence (C(T)) values and a measured quantity of genomic DNA. M. mycoides SC was successfully detected in bronchoalveolar lavage samples obtained from experimentally infected cattle. These TaqMan-based real-time PCR assays will allow for the rapid and specific detection of M. mycoides SC. PMID- 16257144 TI - Analysis of feeding behavior with direct linear transformation. AB - AIM: The sucking pattern of term infants is composed of a rhythmic alteration of expression and suction movements. The aim is to evaluate if direct linear transformation (DLT) method could be used for the assessment of infant feeding. SUBJECT AND METHODS: A total of 10 gnormalh infants and two infants with neurological disorders were studied using DLT procedures and expression/suction pressure recordings. Feeding pattern of seven gnormalh infants were evaluated simultaneously recording DLT and pressures. The other infants were tested non simultaneously. We placed markers on the lateral angle of the eye, tip of the jaw, and throat. The faces of infants while sucking were recorded in profile. The jaw and throat movements were calculated using the DLT procedure. Regression analysis was implemented to investigate the relationship between suction and expression pressures and eye-jaw and eye-throat movement. All regression analyses investigated univariate relationships and adjusted for other covariates. RESULTS: Ten gnormalh infants demonstrated higher suction pressure than expression pressure, and their throat movements were larger than jaw movements. Two infants with neurological problems did not generate suction pressure and demonstrated larger movements in their jaw than throat. The simultaneous measurement (n=7) showed a significant correlation, not only between eye-jaw distance and the expression pressure, but also between eye-throat distance and suction pressure. The change in the eye-jaw distance was smaller than the changes in the eye-throat distance in gnormalh infants (p<0.001). CONCLUSIONS: The DLT method can be used to evaluate feeding performance without any special device. PMID- 16257145 TI - Correlation of haemoglobin-acrylamide adducts with airborne exposure: an occupational survey. AB - This paper reports an occupational hygiene survey of exposure to acrylamide comparing acrylamide haemoglobin adduct measurements with personal air monitoring and glove liner analysis. The air monitoring data showed that exposure to acrylamide was well-controlled with all samples below the UK maximum exposure limit (MEL) of 300 microg/m(3) with mean exposure about one tenth of the MEL. Each worker provided two blood samples approximately 3 months apart. These samples were well correlated (r=0.61) with a slope of 0.74, indicating that exposure was reasonably constant. Mean personal airborne acrylamide levels and mean acrylamide haemoglobin adduct levels were well correlated (r=0.72, N=46) and using the calculated linear correlation, exposure at the MEL would be expected to give rise to a haemoglobin adduct level of 1,550 pmol/g globin. Smoking status did not affect the correlation. There was also a correlation between levels of acrylamide detected on gloves and haemoglobin adduct levels. A combined regression model between haemoglobin adducts, airborne acrylamide and acrylamide glove contamination was significant for both airborne acrylamide and gloves with a regression coefficient of 0.89. The study showed that haemoglobin adduct level was a good biomarker of acrylamide exposure which correlated to both inhaled and potentially skin absorbed acrylamide estimates. There was excellent discrimination between well-controlled occupational levels and environmental levels from diet and smoking, allowing haemoglobin adduct measurement to be used to determine even low level exposures. Due to the complexity of the current methodology, new techniques would be useful in making haemoglobin adducts more widely applicable. PMID- 16257146 TI - Dissociated primary nerve cell cultures as models for assessment of neurotoxicity. AB - Exogenous and endogenous neurotoxins may have poisoning effects on living organisms. Neurotoxic signs can result from human intoxication by substances present in natural ecosystems as pollutants, such as inorganic mercury, cadmium, manganese and lead, or by abnormal accumulation of endogenous compounds, as bilirubin. Dissociated primary nerve cell cultures are powerful models that can be used to evaluate the responses of target cells at the cellular and molecular levels to the deleterious effects of neurotoxic substances. Primary cultures of nerve cells are prepared from either fetal (neurons) or 2-day-old (macroglia and microglia) rat brains, cultured with specific media. Cells can then be used to evaluate the neurotoxic effects of a particular substance. By using cells with different days-in-culture it is possible to mimic and evaluate developmental related modifications. These modifications can comprise morphological changes, cell death by necrosis (release of lactate dehydrogenase, LDH) and apoptosis (nuclear fragmentation), altered neurotransmission (impaired uptake or increased release of glutamate), neuroinflammation (enhanced cytokine production) and the generation of oxidative damage (formation of reactive oxygen species and disruption of glutathione metabolism). Here we describe the methods for nerve cell cultures, as well as some of the procedures that can be used to assess neuronal and glial cytotoxicity induced by different neurotoxins. PMID- 16257147 TI - Reference values for ethylenethiourea in urine in Northern Italy: results of a pilot study. AB - This study was carried out to define reference values for urinary ethylenethiourea (ETU) in the Northern Italy population and to identify the sources of exposure. Ninety-five healthy subjects were selected. A spot urine sample was collected in the morning, and analyzed using GC/MS in the EI/SIM mode. Thirty-nine subjects showed urinary ETU concentrations lower than the limit of detection (LOD, 0.4 microg/g creatinine), and the remainders ETU concentrations ranging from 0.5 to 11.6 microg/g creatinine. No correlation was shown between smoke or alcohol intake and urinary ETU concentrations. Based on data on ethylene bis-dithiocarbamate (EBDC) concentrations in food, we estimated a total EBDCs intake of 31.7-50.1 microg/day. These values are largely below the ADIs, but explain the presence of small amounts of ETU in the urine samples we have analyzed. Finally, it was estimated that the mean ETU in urine in the Italian general population is 0.6-0.8 microg/g creatinine, with a 95th percentile of 4.5 5.0 microg/g creatinine. These values can be used as reference, to compare the results of biological monitoring activities carried out on EBDCs occupationally and environmentally exposed populations. PMID- 16257148 TI - Biomarker of pyrethrum exposure. AB - Pyrethrum as well as synthetic pyrethroids like allethrin, resmethrin, phenothrin, tetramethrin, cyfluthrin, cypermethrin, deltamethrin or permethrin are among the insecticides most often used worldwide. With a sensitive and valid gas-chromatographic-high resolution mass spectrometric method, it is possible to detect all pyrethrum and pyrethroid metabolites in one analytical run. Thus, for the first time a background level of trans-chrysanthemumdicarboxylic acid (CDCA) in urine (95th percentile: 0.15 microg/l) as a characteristic, e.g. for a pyrethrum exposure was found. Following a pyrethrum exposure lasting 1 day, CDCA was found in 27 out of 30 subjects with concentrations going up to 54 microg/l urine (mean: 1.1+/-4.4 microg/l). To obtain information about the elimination kinetics of pyrethrum in humans, urinary excretion of CDCA was investigated in three volunteers after oral intake of 0.3mg pyrethrin I. CDCA was detected during the first 36 h after intake with elimination being most rapid during the first 4h (mean elimination half-life: 4.2h). PMID- 16257149 TI - Effect of post-mating GnRH analogue (buserelin) treatment on PGF2alpha release in ewes and ewe lambs. AB - The objectives of this study were to determine the effects of buserelin or saline treatment on ovarian function (Experiment 1), plasma PGFM concentrations and oxytocin stimulated prostaglandin F(2alpha) (PGF(2alpha)) release (Experiment 2) in ewe lambs and ewes. Welsh Halfbred ewes (n=26) and ewe lambs (n=24) were mated to vasectomised rams at synchronised oestrus and on Day 12 post-mating each animal was injected intramuscularly either normal saline or 4 microg buserelin. In Experiment 1, plasma progesterone and oestradiol concentrations were determined in samples collected by jugular venepuncture 1h before and at 0, 2, 4, 6, 8, 24, 48 and 72 h after treatment (n=7 per treatment group). Progesterone concentrations increased (P<0.05) from 2 to 8h after buserelin treatment and returned to basal levels after 72 h, whereas oestradiol concentrations were maximal at 2h post-treatment and returned to basal levels after 24h (P<0.05). Oestradiol concentrations were lower (P<0.05) in buserelin-treated animals than controls at 72 h post-treatment. Basal and post-treatment progesterone concentrations were greater (P<0.05) in ewes than in ewe lambs but oestradiol levels were similar for both age groups. Ovulation rate, determined by laparoscopy on Day 14, was similar for both age groups (ewes 1.1; ewe lambs 1.0). Buserelin treatment induced accessory corpora lutea in ewes (4/7; 57%) but not in ewe lambs (0/7; 0%). In the Experiment 2, plasma PGFM concentrations were determined in samples collected at 20-min intervals for 6h on Day 14 and at 20 min intervals for 1h before and at 10-min intervals for 1h and then at 20-min intervals for a further 3h period after an intravenous injection of oxytocin (1IU/kg body weight) on Day 15 post-oestrus. In this experiment there were five ewe lambs and six ewes per treatment group. There was no effect of buserelin treatment or age on basal PGFM concentrations on either Day 14 or 15. Although peak PGFM concentrations tended to be lower in buserelin-treated animals, the difference was not significant (P>0.05). However, peak duration following oxytocin challenge on Day 15 post-mating was shorter (P<0.05) in control ewes compared with control ewe lambs. In conclusion, buserelin treatment given on Day 12 post-oestrus enhances luteal function more in ewes than ewe lambs and after a transitory increase, reduces oestradiol concentrations in both ewes and ewe lambs. However, buserelin treatment does not significantly attenuate the luteolytic signal. PMID- 16257150 TI - Combination of anabolic and antiresorptive agents for the treatment of osteoporosis. AB - Combination therapy strategies for osteoporosis are reviewed in this article. Regimens including two antiresorptives agents increase BMD more than single therapy alone, but they have not proved to reduce fracture risk. The alternative of combining antiresorptives together with anabolics has gained more interest. There are, however, many questions still pending, as which antiresorptive should be used, and the appropriate sequence of the treatments. It seems that bisphosphonates can blunt the anabolic effect of parathyroid hormone (PTH) when given simultaneously. Instead, when PTH follows the antiresorptive treatment the anabolic effect seems not to be blunted. Finally, other possible approach would be to initiate therapy with an anabolic agent and maintain the effect with any antiresorptive if necessary. PMID- 16257151 TI - Randomized clinical trial comparing conjugated equine estrogens and isoflavones in postmenopausal women: a pilot study. AB - OBJECTIVE: The aim of this study was to evaluate the effects of isoflavone on the climacteric symptoms (Kupperman Menopausal index), vaginal pH, vaginal cytology (vaginal maturation index) and endometrium (evaluated by ultrasound and biopsy) in postmenopausal women. METHODS: It was a single-center, 6-month, randomized, double-blind, estrogen-controlled trial. Seventy-nine women were randomly assigned to one of the two treatment groups: isoflavone (n=40): 300 mg of the standardized soy extract with a medium dose of 120 mg isoflavones/day as glycoside and aglycone (60 mg twice a day), or estrogen (n=39): one capsule of 0.625 mg conjugated equine estrogens and other capsule with glucose 0.625 mg (placebo). RESULTS: After treatment, there was a decrease in the symptomatology in both estrogen and isoflavone groups. There was a significant decrease in vaginal pH, an increase in superficial vaginal cells and endometrium proliferation after 3 and 6 months of treatment in the estrogen group, but no differences were observed in the isoflavone group for these variables. CONCLUSIONS: We concluded that the daily standardized soy extract with 120 mg isoflavones' effect on symptoms was similar to that from estrogen. Soy isoflavone has no effect on endometrium and vaginal mucosa during the treatment. PMID- 16257152 TI - Exploring factors influencing osteoporosis prevention and control: a qualitative study of Iranian men and women in Australia. AB - OBJECTIVE: To explore the understanding of osteoporosis risk factors and barriers that influence osteoporosis prevention and control in order to better inform the development of culturally appropriate osteoporosis preventive strategies. METHODS: This qualitative study was based on data collected from 5 focus groups, 10 individual interviews and 3 opportunistic group discussions of Iranian men and women living in Australia. A thematic analysis approach was used and data were managed with a computer-assisted program. Data were collected over 1 year in community settings chosen by participants. RESULTS: The participants had insufficient understanding of osteoporosis risk factors and its implications for their health as they age. There were minor misconceptions about the causes of the disease and how it could be prevented and controlled. Many participants confused osteoporosis and osteoarthritis. Where they did indicate some knowledge of preventive actions, they perceived many obstacles to their undertaking preventive behaviours. CONCLUSIONS: These data suggest that factors adversely influencing osteoporosis prevention and control have their root in misconceptions about osteoporosis and its risk factors, and psychosocial and structural barriers to taking preventive actions. These findings provide insights into barriers that need to be overcome if preventive strategies and intervention programs can be developed to reduce the overall burden of osteoporosis in the community. PMID- 16257153 TI - Mathematical modelling of in situ and in vitro efflux of ciprofloxacin and grepafloxacin. AB - The efflux process due to p-glycoprotein-like mechanisms of ciprofloxacin (CIP) and grepafloxacin (GRX) has been studied "in situ" in rats and "in vitro" in Caco 2 cells. The results were modelled by a curve fitting procedure which allowed the characterization of the passive (Pd) and carrier mediated parameters (Vm and Km) from the raw data without initial velocities estimation. CIP absorption in rat was characterized as a passive diffusion at the assayed concentrations. Although the involvement of an efflux transporter cannot be ruled out, its relevance in the transport of the fluoroquinolone is negligible. In GRX absorption, an efflux process is implicated and it is detected in both absorption models. GRX permeability depends on the intestinal segment, reflecting the previously reported different expression level of the efflux transporters along the gut in rat. A first attempt to correlate the "in vitro" and the "in situ" data has been done. The mathematical model has been constructed using very simplistic assumptions and it will require further refinement but, nevertheless, the results are promising and demonstrate that a good modelling approach helps to identify the system critical parameters and how the system behaviour change when the parameters are modified as it happens when we move from the "in vitro" to the "in situ" level. Predicted versus experimental permeability values show a good correlation, demonstrating that the relevance of the secretion process "in situ" in rat can be predicted from the "in vitro" cell results. PMID- 16257154 TI - Formulation design of a novel fast-disintegrating tablet. AB - As our society is becoming increasingly aged, the development of an appropriate dosage form for the elderly patients is mostly desirable. A novel fast disintegrating tablet was investigated in this study as a user-friendly dosage form for the aged. Advantages of this formulation have sufficient hardness and can be manufactured by commonly used equipment. Saccharides can be divided into high- and low-compressibility categories, and an appropriate material for fast disintegrating tablets was created by taking advantage of this fact. To improve the compressibility of low-compressibility saccharides, particle modification was conducted by coating and granulating a low-compressibility saccharide with a high one to enable the production of a fast-disintegrating tablet. Another discovery was that the high-compressibility saccharide used as a binder solution was present in an amorphous state after the granulation process. The crystal change from amorphous to crystal state intentionally by a conditioning process after compression enabled to increase tablet hardness by strengthening adhesion between particles. The conditioning process made it possible to achieve sufficient hardness while maintaining the fast disintegration time. As a result, this fast disintegrating tablet that can be manufactured by commonly used equipment, can be used for the dosing of a wide range drugs. PMID- 16257155 TI - Effect of beta-sitosterol concentration and high pressure homogenization on the chlorhexidine release from vesicular gels. AB - Previous studies have confirmed that the phase transition of vesicular gels of hydrogenated phospholipids to the less ordered fluid vesicular state was induced by the increase of the beta-sitosterol ratio in the whole gel system and consequently in the lipid bilayer. The purpose of the present study was to evaluate the influence of the beta-sitosterol portion in the lipid bilayer and the effect of high pressure homogenization on the structural characteristics of the prepared gel systems. In addition the influence of beta-sitosterol on the consequent chlorhexidine release from the obtained vesicles and liposomes was also examined. Lipid mixtures were prepared from different molar ratios of lecithin:sterol components (90:10-65:35 mol%). The obtained mixtures were hydrated with the aqueous solution of chlorhexidine digluconate in order to achieve a 30% (w/w) final concentration of the lipid mixtures and a 4% (w/w) concentration of the drug. One portion of the resultant multilamellar vesicles was homogenized by using high pressure. To characterize the homogenized and non homogenized systems, transmission electron microscopy of the freeze-fractured samples and differential scanning calorimetry (DSC) were carried out. A vertical type diffusion cell was applied to determine the amount of released chlorhexidine digluconate. Along with the increase in beta-sitosterol concentration, the fluidity of the membrane as well as its permeability also increased. The increased permeability--caused by the higher beta-sitosterol concentration--and the high pressure homogenization, which increased the dispersity and therefore the surface area, enabled a higher amount of chlorhexidine to be released. The increase of drug release was more pronounced in the case of samples prepared with high pressure homogenization. PMID- 16257156 TI - Nasal administration of carbamazepine using chitosan microspheres: in vitro/in vivo studies. AB - The nasal route is used both for local therapies and, more recently, for the systemic administration of drugs, as well as for the delivery of peptides and vaccines. In this study the nasal administration of Carbamazepine (CBZ) has been studied using microspheres constituted by chitosan hydrochloride (CH) or chitosan glutamate (CG). Blank microspheres were also prepared as a comparison. The microspheres were produced using a spray-drying technique and characterized in terms of morphology (scanning electron microscopy, SEM), drug content, particle size (laser diffraction method) and thermal behaviour (differential scanning calorimetry, DSC). In vitro drug release studies were performed in phosphate buffer (pH 7.0). In vivo tests were carried out in sheep using the microparticles containing chitosan glutamate, chosen on the basis of the results of in vitro studies. The results were compared to those obtained after the nasal administration of CBZ (raw material) alone. For the evaluation of in vivo data statistical analysis was carried out using the unpaired t-test. Spray-drying was a good technique of preparation of CBZ-loaded microspheres. The loading of the drug into the polymeric network always led to an increase in the dissolution rate compared to CBZ raw material. The microspheres obtained using chitosan glutamate had the best behaviour both in vitro and in vivo. They increased the drug concentration in the serum when compared to the nasal administration of the pure drug (Cmax 800 and 25 ng/ml for microspheres and pure drug, respectively). The results obtained indicate that the loading of CBZ in chitosan glutamate microspheres increases the amount of the drug absorbed through the nose. PMID- 16257158 TI - Chemoprevention and cytotoxic effect of Bauhinia variegata against N nitrosodiethylamine induced liver tumors and human cancer cell lines. AB - The chemopreventive and cytotoxic effect of ethanol extract of Bauhinia variegata (EBV) was evaluated in N-nitrosodiethylamine (DEN, 200 mg/kg) induced experimental liver tumor in rats and human cancer cell lines. Oral administration of ethanol extract of Bauhinia variegata (250 mg/kg) effectively suppressed liver tumor induced by DEN as revealed by decrease in DEN induced elevated levels of serum glutamate pyruvate transaminase (SGPT), serum glutamate oxaloacetate transaminase (SGOT), alkaline phosphatase (ALP), total bilirubin, gamma glutamate transpeptidase (GGTP), lipid peroxidase (LPO), glutathione peroxidase (GPx) and glutathione S-transferase (GST). The extract produced an increase in enzymatic antioxidant (superoxide dismutase and catalase) levels and total proteins when compared to those in liver tumor bearing rats. The histopathological changes of liver samples were compared with respective controls. EBV was found to be cytotoxic against human epithelial larynx cancer (HEp2) and human breast cancer (HBL-100) cells. These results show a significant chemopreventive and cytotoxic effect of ethanol extract of Bauhinia variegata against DEN induced liver tumor and human cancer cell lines. PMID- 16257157 TI - In vivo study of liposomes as drug carriers to oral mucosa using EPR oximetry. AB - The purpose of this study was to select the best types of liposomes for use as drug carriers for topical treatment of oral mucosal lesions. Electron paramagnetic resonance (EPR) oximetry, using the paramagnetic probe lithium phthalocyanine, was used in vivo to measure the effects of a hyperemic drug, benzyl nicotinate (BN) which was incorporated into liposomes of varying size and composition. The liposomes were made from either hydrogenated or non-hydrogenated soy lecithin and mixed with polymethyl methacrylate ointment for application. EPR oximetry was used to measure the partial pressure of oxygen (pO2) in the oral mucosa before and after application of liposomes. It was found that the most pronounced changes of pO2 in oral mucosa and also the longest action of the drug occurred after the topical application of BN in multi-lamellar liposomes made from hydrogenated soy lecithin (p<0.0001). When these liposomes were applied to oral mucosa over 3 successive days it was found that pO2 increased the most on the first day, the effect gradually decreased following application on the second and third days. The duration of the resulting hyperemia was the longest on the second day (p<0.01). Among the examined carriers, multi-lamellar liposomes made from hydrogenated soy lecithin appear to be the most appropriate for local drug delivery to oral mucosa. PMID- 16257159 TI - Antimicrobial activities of bharangin from Premna herbaceae Roxb. and bharangin monoacetate. AB - The root nodules of Premna herbaceae, which are being used in ayurvedic system of medicine as gantubharangi for curing several ailments, have been studied for antimicrobial activities. The major compound responsible for the biological activity is bharangin, a yellow colored compound extractable with hexane. Bharangin monoacetate prepared by acetylation of bharangin has been investigated along with bharangin for their antimicrobial activity against gram positive and gram negative bacteria and fungi. In general bharangin monoacetate showed more activity with MIC of 3-6 microg/ml when compared with bharangin, which has MIC of 10-25 microg/ml. The enhanced antibacterial activity is attributed to the presence of acetoxyl group in place of hydroxyl group present in the structure of bharangin. The activity is compared with standard gentamycin for bacteria and nystatin for fungi. PMID- 16257160 TI - In vitro antiplasmodial activity of 18 plants used in Congo Brazzaville traditional medicine. AB - Sixty-six extracts of 18 plants commonly used by traditional healers in Congo Brazzaville for the treatment of malaria have been investigated for in vitro antiplasmodial activity. Ethanolic and dichloromethane extracts of 7 among the 18 studied plants were moderately active (10 microg/mlor=4:1 there was a shift in the central tendency of the MIC scatterplot compared with the results of testing 2:1 ratio. As a result, there was a 2-fold dilution increase in the MIC(50) and MIC(90) values, most evident for H. influenzae and BRO-1-producing M. catarrhalis strains. For beta lactamase-positive strains of H. influenzae, the shift resulted in a change in the interpretive result for 3 isolates (1.0%) from susceptible using the reference method (2:1 ratio) to resistant (8/4 microg/mL; very major error) at the 16:1 ratio. In addition, the number of isolates with MIC values at or 1 dilution lower than the breakpoint (4/2 microg/mL) increased from 5% at 2:1 ratio to 32-33% for ratios 14:1 and 16:1. Our results indicate that, for the beta lactamase-positive strains of H. influenzae and M. catarrhalis, the results of the amoxicillin/clavulanate reference 2:1 ratio testing do not accurately represent all the currently licensed formulations. Pharmacokinetic/pharmacodynamic (PK/PD) target attainment might be compromised when higher amoxicillin/clavulanate ratios are used clinically. With a better understanding of PK/PD parameters, reevaluation of the amoxicillin/clavulanate in vitro susceptibility testing should be considered by the standardizing authorities to reflect the licensed formulations and accurately predict clinical outcomes. PMID- 16257169 TI - Baculoviruses-- re-emerging biopesticides. AB - Biological control of agricultural pests has gained importance in recent years due to increased pressure to reduce the use of agrochemicals and their residues in the environment and food. Viruses of a few families are known to infect insects but only those belonging to the highly specialized family Baculoviridae have been used as biopesticides. They are safe to people and wildlife, their specificity is very narrow. Their application as bioinsecticides was limited until recently because of their slow killing action and technical difficulties for in vitro commercial production. Two approaches for the wider application of baculoviruses as biopesticides will be implemented in future. In countries where use of genetically modified organisms is restricted, the improvements will be mainly at the level of diagnostics, in vitro production and changes in biopesticide formulations. In the second approach, the killing activity of baculoviruses may be augmented by genetic modifications of the baculovirus genome with genes of another natural pathogen. It is expected that the baculoviruses improved by genetic modifications will be gradually introduced in countries which have fewer concerns towards genetically modified organisms. PMID- 16257170 TI - Men considering a hypothetical treatment for prostate cancer: a comparison to patients. AB - BACKGROUND: In facilitating informed decision making about PSA screening, men are often asked to consider the potential consequences of the test, including a diagnosis of prostate cancer and how they would want to be treated. However, there is no empirical evidence thus far demonstrating that men are able to consider this hypothetical situation in a realistic manner. PURPOSE: To compare the features (attributes) of treatments that are important to non-patient men considering a hypothetical diagnosis of prostate cancer with those deemed important to men actually diagnosed with early-stage disease. METHODS: Two groups of men went through a decision aid interview to help them choose between treatment options for early-stage prostate cancer: non-patient men who imagined themselves to be diagnosed with the disease, and newly diagnosed patients. During the interview participants identified features of the treatment and disease that were important to their decisions. RESULTS: The percentage of non-patients and patients that thought particular attributes were important was correlated: r (33) = 0.77, p < 0.01. The effects on bladder and bowel functioning were considered important to >or=50% of each group. In addition to the 22 attributes initially presented, 49% of non-patients and 67% of patients identified additional attributes as being important to their decision. Eight (42%) of the 19 additional attributes were identified by non-patients and patients alike. CONCLUSIONS: The group of non-patient men provided a close approximation to the group of newly diagnosed men with respect to the attributes identified as being important to their treatment decisions for early-stage prostate cancer, suggesting that the consideration of what is important to the decision by non-patient men is realistic. PMID- 16257171 TI - [Peripheral nerve catheterization]. PMID- 16257172 TI - A genome-wide, end-sequenced 129Sv BAC library resource for targeting vector construction. AB - The majority of gene-targeting experiments in mice are performed in 129Sv-derived embryonic stem (ES) cell lines, which are generally considered to be more reliable at colonizing the germ line than ES cells derived from other strains. Gene targeting is reliant on homologous recombination of a targeting vector with the host ES cell genome. The efficiency of recombination is affected by many factors, including the isogenicity (H. te Riele et al., 1992, Proc. Natl. Acad. Sci. USA 89, 5128-5132) and the length of homologous sequence of the targeting vector and the location of the target locus. Here we describe the double-end sequencing and mapping of 84,507 bacterial artificial chromosomes (BACs) generated from AB2.2 ES cell DNA (129S7/SvEvBrd-Hprtb-m2). We have aligned these BACs against the mouse genome and displayed them on the Ensembl genome browser, DAS: 129S7/AB2.2. This library has an average insert size of 110.68 kb and average depth of genome coverage of 3.63- and 1.24-fold across the autosomes and sex chromosomes, respectively. Over 97% of the mouse genome and 99.1% of Ensembl genes are covered by clones from this library. This publicly available BAC resource can be used for the rapid construction of targeting vectors via recombineering. Furthermore, we show that targeting vectors containing DNA recombineered from this BAC library can be used to target genes efficiently in several 129-derived ES cell lines. PMID- 16257173 TI - Teratogenic effects of static magnetic field on mouse fetuses. AB - A short period of exposure of pregnant mice to a strong static magnetic field of 400 mT -- 8000 times that of the earth -- in a dorso-ventral direction had teratogenic effects on developing fetuses. Fetuses were exposed to the static magnetic field in utero for 6 min on 1 day from 7.5 to 14.5 days of pregnancy. Exposed and control groups consisted of 10 pregnant mice each; thus 160 animals were used in total. Various malformations were observed in 15.1%, 13.4%, 15.8%, 16.7%, 20.8%, 24.3%, 24.4%, and 14.1% of fetuses exposed on days 7.5, 8.5, 9.5, I0.5, 11.5, 12.5, 13.5, and 14.5 of pregnancy, respectively. Types of malformations were polydactylism, abdominal fissure, fused rib, vestigial 13th rib, lumbar rib, brain hernia, and curled tail, while only a low incidence (up to 2.8%) of curled tail was detected in control group. These deformations apparently caused by SMF exposure but the effect did not reflect so-called exposure period specificity. PMID- 16257175 TI - Functional role of entorhinal cortex in working memory processing. AB - Our learning and memory system has the challenge to work in a world where items to learn are dispersed in space and time. From the information extracted by the perceptual systems, the learning system must select and combine information. Both these operations may require a temporary storage where significance and correlations could be assessed. This work builds on the common hypothesis that hippocampus and subicular, entorhinal and parahippocampal/postrhinal areas are essential for the above-mentioned functions. We bring up two examples of models; the first one is modeling of in vivo and in vitro data from entorhinal cortex layer II of delayed match-to-sample working memory experiments, the second one studying mechanisms in theta rhythmicity in EC. In both cases, we discuss how cationic currents might be involved and relate their kinetics and pharmacology to behavioral and cellular experimental results. PMID- 16257174 TI - The early embryo loss caused by 2,3,7,8-tetrachlorodibenzo-p-dioxin may be related to the accumulation of this compound in the uterus. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a reproductive and developmental toxicant that can alter endocrine status, leading to decreased fertility and altered embryonic development; however, there are limited reports on TCDD toxicity during early pregnancy. In the present study, pregnant and pseudopregnant NIH mice were exposed to TCDD orally (2, 50 and 100 ng/kg body weight) during early gestation (days 1-8), pre-implantation stages (days 1-3), and peri-implantation to early post-implantation stages (days 4-8). TCDD concentration in uterus, liver, kidney, brain and fat on day 9 of pregnancy was monitored by an aryl hydrocarbon receptor (Ah receptor, AhR)-mediated LacZ reporter system in yeast. Results showed that the number of implanted embryos was significantly reduced on day 9 of gestation by 50 and 100 ng/kg TCDD exposure. The number of implantation sites was lower for animals exposed to TCDD on days 1 3 versus those exposed during days 4-8. Decidualization in pseudopregnant mice was also inhibited by TCDD exposure. TCDD concentrations as low as 2 ng/kg significantly decreased serum progesterone levels but had no effect on serum estradiol. TCDD level in the uterus was equal to levels in the liver, but lower than the fat tissue. These results suggest that TCDD sensitivity might be attributed its local accumulation in the uterus. PMID- 16257176 TI - Firing rate modulation: a simple statistical view of memory trace reactivation. AB - Memory trace reactivation in hippocampal ensembles during sleep has been suggested as a coordinating mechanism for consolidation of new memories. Here we propose a simple statistical scheme allowing analysis of the reactivation of firing rate modulations, with a well-defined null hypothesis. This method allowed reliable detection of ensemble reactivation across three experimental settings. Reactivation of firing rate modulations mirrors several properties of commonly studied reactivation measures: it is stronger during hippocampal sharp waves, and decays over a period of 10-20 min. Moreover, in some conditions, firing rate reactivation covaries with reactivation of cell pair cross-correlations, suggesting the two phenomena reflect similar processes. We propose an attractor network model, with pre-wired attractors, in which experience selects and primes some attractors. Priming occurs by either experience dependent synaptic plasticity or changes in neuronal excitability. Primed attractors are more likely to activate in the following sleep, inducing reactivation of both rates and cross correlations. PMID- 16257177 TI - Investigating the genetic basis of susceptibility to rheumatoid arthritis. AB - The challenges in identifying genetic polymorphisms that influence the susceptibility to rheumatoid arthritis are the same as those faced in most complex diseases; genetic and phenotypic heterogeneity, an unknown number of loci presumed to have small genetic effects, non-genetic modifying effects that have yet to be fully characterised and a history of unconfirmed genetic associations. Despite the difficulties, the chronic nature of the disease, incomplete efficacy of existing therapies and resultant heavy healthcare burden for the developed world in managing patients with this condition, mean that an understanding of the genetic basis of disease susceptibility, severity and response to therapy is keenly sought. Many linkage and association studies have been carried out and in this article the results of linkage studies are summarised. Recently a number of convincing candidate genes have begun to emerge and an update has been provided for three of these: PTPN22, CTLA-4 and MIF. PMID- 16257178 TI - Redefining the HLA and RA association: to be or not to be anti-CCP positive. AB - Rheumatoid Arthritis (RA) is the most common chronic inflammatory joint disease. The overall prevalence is 1% and in people older than 60 it is more than 2%. RA has auto-immune features: auto-antibodies against the Fc part of IgG, so-called Rheumatoid Factor (RF) are found more often in RA patients and more recently RA specific auto-antibodies directed against Cyclic Citrunilated Peptides (CCP) have been discovered. Based on twin studies the contribution of genetic factors to the pathogenesis has been estimated to be about 60%. The main genetic contribution (about 40%) comes from the HLA complex. An association between HLA-DR4 and RA was already documented almost 30 years ago. This association was more prominent for severe forms of the disease. Because more HLA-DRB1 alleles appeared to be associated with RA and the products of these alleles shared a 5AA sequence in a peptide-binding pocket the so-called Shared Epitope (SE) hypothesis was formulated, the prediction being that these DRB1 molecules would bind an RA inducing peptide(s). Thus far however such (a) peptides remain elusive. Because the risk for RA associated with different but SE-identical DRB1 alleles varies considerably this SE can also not be the whole explanation for the HLA contribution to RA susceptibility/severity. A modified SE has been postulated and a role for DQ has been postulated. There is also evidence for a contribution of non-class II genes to susceptibility. About 5 years ago we have reported that certain HLA-DRB1 alleles are associated with protection from (severe) RA. The products of these alleles carry instead of the SE sequence another peptide anchor region consisting of the amino acid DERAA. In a large prospective cohort study we showed recently that these alleles indeed confer (dominant) protection both against developing RA and a severe course of the disease. This protection was observed both in the presence and the absence of SE susceptibility alleles. We are presently exploring the hypothesis that this protection is mediated by regulatory T cells reactive with the DERAA epitope. An obvious way to unravel the apparently complex association between HLA and RA is to reduce the heterogeneity of this multifactorial disease. Recently, we have discovered that SE positive DRB1 alleles are exclusively associated with CCP positive RA. The previously reported association with RF positive RA appeared to be secondary to the association with anti-CCP pos. RA. This was the case both for the association found for susceptibility and severity. Interestingly, DRB1*03 was exclusively associated with anti-CCP neg. RA. Because recent evidence puts the immune response against citrunilated proteins (CCP) as prime suspect for disease induction and progression in this subgroup of RA these observations are a big leap forwards in solving the HLA-RA puzzle. PMID- 16257179 TI - Naive transgenic T cells expressing cartilage proteoglycan-specific TCR induce arthritis upon in vivo activation. AB - Proteoglycan (PG)-induced arthritis (PGIA), a murine model for rheumatoid arthritis (RA), is driven by antigen (PG)-specific T and B cell activation. In order to analyze the pathogenic role of antigen-specific T cells in the development of autoimmune arthritis, we have generated a transgenic (Tg) mouse. The CD4(+) T cells of this TCR-5/4E8-Tg line express a functional T cell receptor (TCR) composed of the Valpha1.1 and Vbeta4 chains with specificity for the dominant arthritogenic T cell epitope of human cartilage PG. Adoptive transfer of naive TCR-5/4E8-Tg cells induced arthritis with severe clinical symptoms in syngeneic immunodeficient BALB/c.RAG2(-/-) mice. In vivo activation of TCR-5/4E8 Tg CD4(+)Vbeta4(+) cells with cartilage PG seemed to be critical for arthritis induction. Arthritis never developed after transfer of naive wild-type cells. The arthritis was characterized as a chronic progressive disease with intermittent spontaneous exacerbations and remissions. Inflamed joints showed extensive cartilage damage and bone erosions leading to massive ankylosis in peripheral joints. These PG epitope-specific TCR-5/4E8-Tg mice can be valuable research tools for studying antigen-driven T cell regulation in arthritis, and migration of T cells to the joints. In addition the model may be used for the development of immune modulating strategies in T cell-mediated autoimmune diseases. PMID- 16257180 TI - Mechanism of induction of muscle protein degradation by angiotensin II. AB - Angiotensin I and II have been shown to directly induce protein degradation in skeletal muscle through an increased activity and expression of the ubiquitin proteasome proteolytic pathway. This investigation determines the role of the nuclear transcription factor nuclear factor-kappaB (NF-kappaB) in this process. Using murine myotubes as a surrogate model system both angiotensin I and II were found to induce activation of protein kinase C (PKC), with a parabolic dose response curve similar to the induction of total protein degradation. Activation of PKC was required for the induction of proteasome expression, since calphostin C, a highly specific inhibitor of PKC, attenuated both the increase in total protein degradation and in proteasome expression and functional activity increased by angiotensin II. PKC is known to activate I-kappaB kinase (IKK), which is responsible for the phosphorylation and subsequent degradation of I kappaB. Both angiotensin I and II induced an early decrease in cytoplasmic I kappaB levels followed by nuclear accumulation of NF-kappaB. Using an NF-kappaB luciferase construct this was shown to increase transcriptional activation of NF kappaB regulated genes. Maximal luciferase expression was seen at the same concentrations of angiotensin I/II as those inducing protein degradation. Total protein degradation induced by both angiotensin I and II was attenuated by resveratrol, which prevented nuclear accumulation of NF-kappaB, confirming that activation of NF-kappaB was responsible for the increased protein degradation. These results suggest that induction of proteasome expression by angiotensin I/II involves a signalling pathway involving PKC and NF-kappaB. PMID- 16257181 TI - Oncogenic K-Ras down-regulates Rac1 and RhoA activity and enhances migration and invasion of pancreatic carcinoma cells through activation of p38. AB - Activating mutations in the K-ras gene are genetic alterations frequently found in human carcinomas, particularly in pancreatic adenocarcinomas. Mutation of the K-ras gene is thought to be an early and important event in pancreatic tumor initiation, but the precise role of the mutant K-Ras proteins in neoplastic progression is still unknown. In the present study, we have characterized the influence of oncogenic K-Ras on the phenotype and on the signal transduction of epitheloid PANC-1 pancreatic carcinoma cells by generating PANC-1 cell clones, which stably express EGFP(enhanced green fluorescent protein)-K-Ras (V12). EGFP-K Ras (V12)-expressing cells exhibited a more fibroblastoid cellular phenotype with irregular cell shape and disorganized cytokeratin filaments. Moreover, these cells showed a marked enhancement of their migratory and invasive properties. Stable expression of EGFP-K-Ras (V12) down-regulated the activity of Rac1 and RhoA, resulting in reduced subcortical actin filaments and stress fibers, which might contribute to the epithelial dedifferentiation. Characterization of the activity of mitogen-activated protein kinases revealed that EGFP-K-Ras (V12) enhanced the activity of p38, but did not affect the activities of the Raf/MEK/ERK cascade and JNK. While inhibition of either MEK or JNK activity had no effect on EGFP-K-Ras (V12)-induced migration, inhibition of p38 activity markedly reduced EGFP-K-Ras (V12)-induced migration. Collectively, the results suggest that oncogenic K-Ras enhances the malignant phenotype and identify the mitogen-activated protein kinase p38 as a target to inhibit oncogenic K-Ras induced pancreatic tumor cell migration. PMID- 16257182 TI - Color vision and contrast sensitivity in epilepsy patients treated with initial tiagabine monotherapy. AB - The purpose of the study was to determine whether the use of a GABAergic antiepileptic drug (AED), tiagabine, affects color vision and contrast sensitivity. Twenty newly diagnosed patients with partial epilepsy (aged 19-72 years), receiving tiagabine as their initial monotherapy for 5-41 months were examined. Color vision was examined with the Standard Pseudoisochromatic Plates 2 (SPP2), with the Farnsworth-Munsell 100 Hue Test (FM100) and with the Color Vision Meter 712 (CVM) anomaloscope. Contrast sensitivity was measured with the Pelli-Robson letter chart. Three patients excluded from the color vision evaluation for congenital red-green color vision defects. Seven out of 17 patients (41%) had acquired color vision deficit examined with the FM100. The CVM anomaloscope revealed minor defects in two patients. Contrast sensitivity function was within normal ranges. The present study suggests that AED therapy with tiagabine, like with other established and newer AEDs may interfere with color perception. PMID- 16257183 TI - The relevance of neurosteroids to clinical psychiatry: from the laboratory to the bedside. AB - Neurosteroids are important neuroactive molecules with suggested central involvement in several neurophysiological and psychiatric disease processes. The discovery of neurosteroids followed the revelation that the brain exhibited the capacity to synthesize its own steroids in situ and thus be a potential site of steroidogenesis. In contrast to some steroids that exhibit traditional genomic steroid actions, most neurosteroids appear to regulate neuronal function by means of "non-genomic" mechanisms influencing neuronal excitability. Neurosteroids are synthesized either from CNS cholesterol or from peripheral steroid precursors and exhibit a wide range of modulatory effects on neurotransmitter receptor activity, most notably at the gamma-aminobutyric acid A (GABA(A)) receptor. Neurosteroids play an important role in neurodevelopment and neuroprotective effects, many aspects of which may have particular applicability to psychiatric disorders including various gender differences. Neurosteroids appear to be relevant to the pathophysiology and pharmacological treatment of many psychiatric disorders including the most notable mood and anxiety disorders, but also psychotic, childhood, eating, dementia, stress and postpartum disorders. It has been suggested that neurosteroids may become potential targets for pharmacological intervention in the future with further neurosteroid investigation contributing to a more comprehensive understanding of human behavior and psychopathology. PMID- 16257184 TI - Potent protection of ferulic acid against excitotoxic effects of maternal intragastric administration of monosodium glutamate at a late stage of pregnancy on developing mouse fetal brain. AB - The present study was conducted to investigate a possible protection of ferulic acid against excitotoxic effects of maternal intragastric (ig) administration of monosodium glutamate (MSG) at a late stage of pregnancy on developing mouse fetal brain. [(3)H]-labeled glutamate was used as radiotracer to study the effect of ferulic acid on distribution of MSG in mouse fetal brain. MSG dissolved in distilled water (2.0 g/kg body weight, 640 kBq of [(3)H]glutamate/mouse, ig) or/and sodium ferulate (SF) (20, 40, 80 mg/kg body weight, ip), was given to pregnant mice at 17-19 days; the distribution of [(3)H] glutamate in the mouse fetal brains was measured at 30, 60, 90, 120 min after administration of MSG or/and SF. Maternal mice were given MSG (1.0, 2.0, 4.0 g/kg body weight, ig) or/and SF (20, 40, 80 mg/kg body weight, ip) simultaneously at 17-19 days of pregnancy, and then behavioral tests and histopathological observations were used to analyze glutamate-induced functional and morphological changes of the brains of their offspring, and Western blot analysis was performed for examining expressions of bcl-2 and caspase-3. The results showed that SF obviously inhibited the uptake of labeled glutamate in fetal brain. In addition, SF countered the effects of MSG on behavior, histopathology, genetic toxicity, and expression of apoptosis-related gene. The results suggest that ferulic acid is a novel competitive N-methyl-D-aspartate (NMDA) receptor antagonist and neuroprotector. In conclusion, maternal administration of ferulic acid has potent protective effects against glutamate-induced neurotoxicity in their filial mice. PMID- 16257185 TI - Collagen type IX and HNK-1 epitope in tears of patients with pseudoexfoliation syndrome. AB - Pseudoexfoliation syndrome (PEX) is an age-related condition, which may cause open-angle glaucoma and has increasing interest since it seems to affect additional human tissues, i.e., cardiovascular tissue, skin, and still lacks elucidated pathogenesis. Collagen type IX and HNK-1 epitope have been considered characteristic constituents of the aqueous humour of PEX patients, since their amounts were increased in PEX aqueous humour compared to normal eyes. Since it has been proposed that the initial manifestations of PEX syndrome occur in conjunctiva, the present study was undertaken to investigate the presence of the same antigens in tears of PEX patients and their possible use as the biochemical markers for early diagnosis. Tears of PEX patients and healthy individuals were subjected to western blotting analysis for various basement membrane components identified in aqueous humour. It was found that collagen type IX and HNK-1 epitope were present in tears, the amount of the former being increased 2.7 times compared to normal (P<0.05), surprisingly high as compared with total protein or lysozyme activity in tears, which were found to be increased in PEX patients about 25% with no statistical differences (P approximately 0.4). The results suggest the possible use of tears' collagen type IX for the diagnosis of PEX syndrome. PMID- 16257187 TI - Rapid T1 measurement via decay-recovery decomposition: applications in fringe field and distributed relaxation experiments. AB - Spin-lattice relaxation time (T(1)) measurements are often time-consuming due to the need to measure the full equilibrium magnetization with a long wait time. However, any magnetization recovery can be decomposed into pure recovery and pure decay components, the latter of which lends itself to a much simpler and faster extraction of T(1). We demonstrate several pulse sequences that accomplish this decomposition experimentally and illustrate its applications in a steady magnetic field gradient, and in materials possessing a broad distribution of T(1). PMID- 16257186 TI - Functional activation imaging in aging and dementia. AB - With life expectancy increasing continuously, the effects of neurodegeneration on brain function are a topic of ever increasing importance. Thus there is a need for tools and models that probe both the functional consequences of neurodegenerative processes and compensatory mechanisms that might occur. As neurodegenerative burden and compensatory mechanisms may change over time, these tools will ideally be applied multiple times over the lifespan. Specifically, in order to elucidate whether brain-activation patterns in Alzheimer's disease (AD) and in healthy aging follow general rules in the context of degeneration and compensation, it is necessary to compare functional brain-activation patterns during different states of neurodegeneration. This article integrates the findings of functional activation studies at different stages of neurodegeneration: in healthy aging, in subjects at high risk of developing dementia, in subjects with mild cognitive impairment (MCI), and in patients suffering from AD. We review existing theoretical models that aim to explain the underlying mechanisms of functional activation changes in aging and dementia, and we propose an integrative account, which allows for different neural response patterns depending on the amount of neuronal damage and the recruitment of compensatory pathways. PMID- 16257188 TI - A REDOR study of diammonium hydrogen phosphate: a model for distance measurements from adsorbed molecules to surfaces. AB - Magic angle spinning NMR techniques can be used to determine the molecular structure of proteins adsorbed onto polymer and mineral surfaces, but the degree to which the orientation of proteins on surfaces can be uniquely determined by NMR is less well understood. In this manuscript, REDOR data obtained from model systems are analyzed with a view to determine the orientation of rare spins coupled to a lattice populated by strongly coupled spin 1/2 nuclei. When the surface is populated by closely spaced spins, the REDOR dephasing of a rare spin on the protein contact point to the surface is under certain circumstances complicated by contributions from homonuclear dipolar interactions between the spins of the lattice. To study multiple spin effects on the dephasing signal in rotational-echo-double-resonance experiments, we carried out a measurement on crystalline diammonium hydrogen phosphate as a model for a spin system with multiple dipolar interactions. Information about the (31)P-(31)P interactions is gathered from the reference measurement in the experiment. To fit the experimental (15)N and (31)P dephasing data well, it was necessary to account for as many as 6 and 8 spins in simulations, respectively. Using a single spin-pair interaction with an unknown distance yielded a good fit to the (31)P data with a distance of 2.7A that is nearly an Angstrom shorter than the shortest distance in the crystal structure. Homonuclear couplings are shown to have a significant effect on the expected dephasing. PMID- 16257189 TI - Multidimensional 2H NMR study of dynamical heterogeneity in polymer nanocomposites. AB - Nanoconfined polymer chains-as can be formed when polymers intercalate into layered inorganic materials-show remarkable bulk properties, many of which are connected to dynamical heterogeneity in the polymeric phase. Microscopically, it appears that slow dynamical modes are associated with the species in direct contact with the surface, with substantially more mobile species only a fraction of a nm away. In the more distant phase, larger angle and moderately fast dynamics (typically, ns-micros) grows in over a broad temperature range not well correlated to bulk phase transitions. In this work, we probe the slowest dynamical modes and apply one- and two-dimensional (2)H exchange NMR experiments to study thin polymer layers intercalated between the flat inorganic faces of fluorohectorite (FH). One sample is created by intercalation of perdeuterated poly(ethylene oxide) into FH, and the second by intercalation of d(3) poly(styrene) into a surface-modified FH. Large-amplitude reorientation of the PEO backbone is substantially hindered in the narrow two-dimensional layers, and reorientation is limited to small-amplitude steps at rates that are largely independent of temperature. Simulations of the two-dimensional exchange experiments suggest that dynamics in nanoconfined polymers is associated with small-angle rotational diffusion. PMID- 16257190 TI - Analysis of porosity in porous silicon using hyperpolarized 129Xe two-dimensional exchange experiments. AB - The porosity in porous silicon was characterized using hyperpolarized (HP) xenon as a probe. HP xenon under conditions of continuous flow allows for the rapid acquisition of xenon NMR spectra that can be used to characterize a variety of materials. Two-dimensional exchange spectroscopy (EXSY) (129)Xe NMR experiments using HP xenon were performed to obtain exchange pathways and rates of xenon mobility between pores of different dimensions within the structure of porous silicon and to the gas phase above the sample. Pore sizes are estimated from chemical shift information and a model for pore geometry is presented. PMID- 16257191 TI - Age-related change in neural processing of time-dependent stimulus features. AB - Aging is associated with changes in automatic processing of task-irrelevant stimuli, and this may lead to functional disturbances including repeated orienting to nonnovel events and distraction from task. The effect of age on automatic processing of time-dependent stimulus features was investigated by measurement of the auditory mismatch negativity (MMN) in younger (18-23) and older (55-85) adults. Amplitude of MMN recorded during a paradigm involving low probability deviation in interstimulus interval (from 500 ms to 250 ms) was found to be reduced in the older group at fronto-central sites. This effect was paralleled by, and correlated to, decreased sensory gating efficiency for component N1 recorded during a separate paradigm involving alternate presentation of auditory stimuli at long (9 s) and short (0.5 s) interstimulus intervals. Further, MMN amplitude was correlated to behavioral performance on a small subset of neuropsychological tests, including the Rey Auditory Verbal Learning Test, within a group of healthy older adults. The results support the hypothesis that aging is associated with declines in automatic processing of time-dependent stimulus features, and this is related to cognitive function. These conclusions are considered in the context of age-related declines in prefrontal cortex function and associated increases in susceptibility to task-irrelevant stimuli. PMID- 16257193 TI - [Management of children with hypercholesterolemia]. AB - The management of children with hypercholesterolemia (HC) depends on the level of premature atherosclerosis - associated risk. Inherited autosomal-dominant forms of HC (Family HC, Familial deficiency of apolipoprotein B, Family combined dyslipidemia) are at high risk of premature cardiovascular disease. These inherited forms of HC need to be systematically screened during childhood in case of family history and require a long term follow-up in order to prevent adult coronary insufficiency. The first recommended therapy consists in dietary intervention. When necessary, treatment with statin can be used from 8 years old. Before this age, acid-binding resins remain the first step treatment. Plant sterol-esters enriched spreads could be an additional useful treatment. PMID- 16257192 TI - Functional characterization of a human organic anion transporter hOAT4 in placental BeWo cells. AB - Human organic anion transporter 4 (hOAT4) belongs to a family of organic anion transporters which play critical roles in the body disposition of clinically important drugs, including anti-HIV therapeutics, anti-tumor drugs, antibiotics, anti-hypertensives, and anti-inflammatories. hOAT4 is expressed in the placenta and kidney. In the current study, we stably transfected hOAT4 into human placental BeWo cells and the functional properties of hOAT4 and its regulation were investigated in these cells. hOAT4-mediated uptake of estrone sulfate, a protypical organic anion for hOAT4, was dose- and time-dependent, and saturable (Km=4.2 microM). The substrate specificity of hOAT4 includes various steroid sulfates, such as beta-estradiol-3,17-disulfate, 17-beta-estradiol-3-sulfate, beta-estradiol-3-sulfate, and dehydroepiandrosterone-3-sulfate (DHEAS), but does not include p-aminohippuric acid (PAH) and tetraethylammonium (TEA). Pre incubation of hOAT4-expressing BeWo cells with phorbol 12-myristate 13-acetate (PMA) and phorbol 12,13-dibutyrate (PDBu), both of which are protein kinase C (PKC) activators, acutely inhibited the transport activity. The inhibition by PDBu resulted in a decreased Vmax without significant affecting the Km. Establishment of hOAT4-expressing BeWo cells provided useful tool for further pharmacological and molecular biological studies of placental transport of organic anions mediated by this carrier. PMID- 16257194 TI - Effect of expectoration on inflammation in induced sputum in alpha-1-antitrypsin deficiency. AB - It is unclear how chronic expectoration influences airway inflammation in patients with chronic lung disease. The aim of this study was to investigate factors influencing inflammation in induced sputum samples, including, in particular, chronic sputum production. Myeloperoxidase, interleukin-8, leukotriene B4 (LTB4), neutrophil elastase, secretory leukoprotease inhibitor (SLPI) and protein leakage were compared in induced sputum samples from 48 patients (36 with chronic expectoration) with COPD (with and without alpha-1 antitrypsin deficiency; AATD), 9 individuals with AATD but without lung disease and 14 healthy controls. There were no differences in inflammation in induced sputum samples from healthy control subjects and from AATD deficient patients with normal lung function but without chronic expectoration (P>0.05). Inflammation in induced sputum from AATD patients with airflow obstruction and chronic sputum expectoration was significantly greater than for similar patients who did not expectorate: Interleukin-8 (P<0.01), elastase activity (P=0.01), and protein leakage (P<0.01). The presence of spontaneous sputum expectoration in AATD patients with airflow obstruction was associated with increased neutrophilic airway inflammation in induced sputum samples. The presence of chronic expectoration in some patients will clearly complicate interpretation of studies employing sputum induction where this feature has not been identified. PMID- 16257195 TI - Metabolic capacity of the diaphragm in patients with COPD. AB - Chronic obstructive pulmonary disease (COPD) is associated with an increased load on the diaphragm. Chronic loading on skeletal muscles results in metabolic changes and fiber-type shifts. Therefore, we investigated whether the load on the human diaphragm imposed by COPD altered oxidative enzyme activity, glycogenolytic enzyme activity and mitochondrial energy generating capacity and efficiency. Biopsies of the diaphragm from COPD patients and control subjects were obtained and activities of L(+)3-hydroxyacylCoA-dehydrogenase (HADH, marker for beta oxidation capacity) and phosphorylase (marker for glycogenolytic capacity) were measured spectrophotometrically. Mitochondrial energy generating capacity was measured by spectrophotometrical and radiochemical methods. Fiber-type distribution was determined electrophoretically. We found that HADH activity was increased with increasing severity of COPD (P=0.05). No change in glycogenolytic enzyme activity was observed. The activity of the mitochondrial respiratory chain complexes III and IV and oxidation of pyruvate was increased with increasing airflow obstruction. These results suggest that in COPD the diaphragm adapts to a higher workload by increasing the oxidative capacity and mitochondrial function. PMID- 16257196 TI - Medical wastes management in the south of Brazil. AB - Medical (clinical) wastes are costly in disposal and carry risks of infection, or physical injury, and of exposure to potentially harmful pharmaceuticals, as well as being aesthetically unacceptable. Technological advances in disposal, together with the introduction of rigorous emission standards for incinerators and similarly stringent control standards for non-burn "alternate" disposal technologies, continue to drive improvements in waste management. Are these improvements attainable in developing countries? Where adequate resources and a robust infrastructure are lacking, investment in advanced disposal technologies may be counterproductive. Developments must be appropriate to and manageable by the communities served; sustainable low-technology approaches may be preferable. There remains a need for affordable technical innovation, as well as underlying political support and international financial and technical assistance, to sustain meaningful improvements in waste management in remote and isolated regions and more generally in developing countries. PMID- 16257197 TI - Automatic fermentation control based on a real-time in situ SIRE biosensor regulated glucose feed. AB - Monitoring and regulation of fermentations is of a paramount industrial and academic importance in order to keep conditions optimal during the entire process. Established techniques employed today include HPLC and spectrophotometry, which both have the disadvantage that broth samples have to be drawn from the fermentor and that they often require sample pre-treatment. The objectives of this study was to design and evaluate a software controlled automatic real-time SIRE biosensor connected to a glucose feed solution pump for in situ based monitoring and regulation of the glucose concentration during a yeast fermentation process. The maximal frequency for the measuring-regulation cycles was 30/h. A 10 mM mean glucose concentration level was successfully maintained within +/-0.013 mM during 60 min fermentations at various concentrations of yeast (10, 20, 40 and 80g/l). The on/off-regulator used caused some expected fluctuations (oscillations) of the glucose concentration around the mean value (+/-0.12 mM at 10 g/l, +/-0.26 mM at 20 g/l, +/-0.51 mM at 40 g/l, and +/-0.99 mM at 80 g/l). A 7-h fermentation process (10 mM glucose and 20 g/l yeast) was successfully monitored and regulated. The obtained measuring data were found to be 8.5-22.9% lower than data obtained with a commercially available spectrophotometric kit. The difference increased linearly (-0.26 mM/h), during the fermentation process and indicated that some clogging of the in situ positioned probe occurred. The speed and the automatisation adaptability of the presented device suggest advantages compared to established techniques. PMID- 16257198 TI - Nucleic acid enzymes. AB - Since the discovery of the first natural ribozyme more than 20 years ago, it has become clear that nucleic acids are not only the static depository of genetic information, but also possess intriguing catalytic activity. The number of reactions catalyzed by engineered nucleic acid enzymes is growing continuously. The versatility of these catalysts supports the idea of an ancestral world based on RNA predating the emergence of proteins, and also drives many studies towards practical applications for nucleic acid enzymes. PMID- 16257199 TI - Synthetic dye decolorization capacity of white rot fungus Dichomitus squalens. AB - The ability to decolorize eight chemically different synthetic dyes (Orange G, Amaranth, Orange I, Remazol Brilliant Blue R (RBBR), Cu-phthalocyanin, Poly R 478, Malachite Green and Crystal Violet) by the white rot fungus Dichomitus squalens was evaluated on agar plates. The fungus showed high decolorization capacity and was able to decolorize all dyes tested, but not to the same extent. Some of the dyes did not limit the decolorization capacity of the strain tested even at a concentration of 2g/l. The presence of the dyes in solid media reduced the mycelial growth rate of D. squalens; a positive correlation was found between the growth rate and the decolorization ability. Decolorization of Orange G and RBBR was studied also in liquid culture, where both dyes caused an enhancement of ligninolytic enzyme and overall hydrogen peroxide production and a decrease of biomass production. RBBR was removed to a higher extent than Orange G. PMID- 16257200 TI - Lactic acid fermentation of food waste for swine feed. AB - This study was conducted to determine the effects of lactic acid bacteria (LAB, Lactobacillus salivarius) inoculation on the microbial, physical and chemical properties of food waste mixture (FWM) stored at ambient temperature (25 degrees C) for 10 and 30 days. A complete pig diet including restaurant food waste, bakery by-product, barley and wheat bran, and broiler poultry litter was amended with LAB at the levels of 0.1%, 0.2%, 0.5% and 1.0% and fermented anaerobically. These treatments were compared with intact FWM before storage and non anaerobically stored FWM. Non-anaerobic storage of FWM showed microbial putrefaction with the loss (P < 0.05) of water and water soluble carbohydrate (WSC) and increases (P < 0.005) in protein and fiber. Anaerobic fermentation of FWM with or without LAB seemed effective in both 10- and 30-day-storage. The addition of LAB inoculants to FWM showed a linear trend (P < 0.05) toward an increase in the number of total and lactic acid bacteria and toward the nutritional improvement with WSC increased and fiber decreased. Long-term (30 days) storage resulted in consistent reduction (P < 0.05) in numbers of total and lactic acid bacteria and pH and showed little change in chemical components, compared with short-term (10 days) storage. On the basis of these results, LAB inoculation improved fermentative characteristics of FWM. Among anaerobic treatments, further WSC increase and NDF reduction did not occur (P > 0.05) when LAB-added levels were over 0.2%. Based on these observations the optimum level of LAB addition to FWM was 0.2%. PMID- 16257201 TI - Privileged structure-based quinazolinone natural product-templated libraries: identification of novel tubulin polymerization inhibitors. AB - A focused quinazolinone natural product-templated library was designed and synthesized. Compounds from this privileged structure-based library were identified as antimitotic agents acting through destabilization of tubulin polymerization. The results suggested that 2 could be a privileged substructure. PMID- 16257202 TI - Design and synthesis of heterocyclic malonyl-CoA decarboxylase inhibitors. AB - We have previously reported the discovery of small molecule inhibitors of malonyl CoA decarboxylase (MCD) as novel metabolic modulators, which inhibited fatty acid oxidation and consequently increased the glucose oxidation rates in the isolated working rat hearts. MCD inhibitors were also shown to improve cardiac efficiency in rat and pig demand-induced ischemic models through the mechanism-based modulation of energy metabolism. Herein, we describe the design and synthesis of a series of novel heterocyclic MCD inhibitors with a preference for substituted imidazole and isoxazole. PMID- 16257203 TI - Structure-based design of novel groups for use in the P1 position of thrombin inhibitor scaffolds. Part 1: Weakly basic azoles. AB - Despite their relatively weak basicity, simple azoles, specifically imidazoles and aminothiazoles, can function as potent surrogates for the more basic amines (e.g., alkyl amines, amidines, guanidines, etc.) which are most often employed as the P1 ligand in the design of noncovalent small molecule inhibitors of thrombin. PMID- 16257204 TI - Generation of potent coagulation protease inhibitors utilizing zinc-mediated chelation. AB - Inhibition of coagulation proteases such as thrombin, fXa, and fVIIa has been a focus of ongoing research to produce safe and effective antithrombotic agents. Herein, we describe a unique zinc-mediated chelation strategy to streamline the discovery of potent inhibitors of fIIa, fXa, and fVIIa. SAR studies that led to the development of selective inhibitors of fXa will also be detailed. PMID- 16257205 TI - Antimalarial activity of 4-(5-trifluoromethyl-1H-pyrazol-1-yl)-chloroquine analogues. AB - The antimalarial activity of chloroquine-pyrazole analogues, synthesized from the reaction of 1,1,1-trifluoro-4-methoxy-3-alken-2-ones with 4-hydrazino-7 chloroquinoline, has been evaluated in vitro against a chloroquine resistant Plasmodium falciparum clone. Parasite growth in the presence of the test drugs was measured by incorporation of [(3)H]hypoxanthine in comparison to controls with no drugs. All but one of the eight (4,5-dihydropyrazol-1-yl) chloroquine 2 derivatives tested showed a significant activity in vitro, thus, are a promising new class of antimalarials. The three most active ones were also tested in vivo against Plasmodium berghei in mice. However, the (pyrazol-1-yl) chloroquine 3 derivatives were mostly inactive, suggesting that the aromatic functionality of the pyrazole ring was critical. PMID- 16257206 TI - Derivatives of tramadol for increased duration of effect. AB - Tramadol is a centrally acting opioid analgesic structurally related to codeine and morphine. Analogs of tramadol with deuterium-for-hydrogen replacement at metabolically active sites were prepared and evaluated in vitro and in vivo. PMID- 16257207 TI - Pyrrolo(iso)quinoline derivatives as 5-HT(2C) receptor agonists. AB - A series of 1-(1-pyrrolo(iso)quinolinyl)-2-propylamines was synthesised and evaluated as 5-HT(2C) receptor agonists for the treatment of obesity. The general methods of synthesis of the precursor indoles are described. The functional efficacy and radioligand binding data for the compounds at 5-HT(2) receptor subtypes are reported. The analogue which showed the highest 5-HT(2C) binding affinity (27, 1.6nM) was found to be successful in reducing food intake in rats. PMID- 16257209 TI - Rounding-up on the TCB Cytokinesis series. PMID- 16257210 TI - Coordinating the events of the meiotic prophase. AB - Meiosis is a specialized type of cell division leading to the production of gametes. During meiotic prophase I, homologous chromosomes interact with each other and form bivalents (pairs of homologous chromosomes). Three major meiotic processes--chromosome pairing, synapsis and recombination--are involved in the formation of bivalents. Many recent reports have uncovered complex networks of interactions between these processes. Chromosome pairing is largely dependent on the initiation and progression of recombination in fungi, mammals and plants, but not in Caenorhabditis elegans or Drosophila. Synapsis and recombination are also tightly linked. Understanding the coordination between chromosome pairing, synapsis and recombination lends insight into many poorly explained aspects of meiosis, such as the nature of chromosome homology recognition. PMID- 16257208 TI - Biocatalytic ammonolysis of (5S)-4,5-dihydro-1H-pyrrole-1,5-dicarboxylic acid, 1 (1,1-dimethylethyl)-5-ethyl ester: preparation of an intermediate to the dipeptidyl peptidase IV inhibitor Saxagliptin. AB - An efficient biocatalytic method has been developed for the conversion of (5S) 4,5-dihydro-1H-pyrrole-1,5-dicarboxylic acid, 1-(1,1-dimethylethyl)-5-ethyl ester (1) into the corresponding amide (5S)-5-aminocarbonyl-4,5-dihydro-1H-pyrrole-1 carboxylic acid, 1-(1,1-dimethylethyl)ester (2), which is a critical intermediate in the synthesis of the dipeptidyl peptidase IV (DPP4) inhibitor Saxagliptin (3). Candida antartica lipase B mediates ammonolysis of the ester with ammonium carbamate as ammonia donor to yield up to 71% of the amide. The inclusion of Ascarite and calcium chloride as adsorbents for carbon dioxide and ethanol byproducts, respectively, increases the yield to 98%, thereby offering an efficient and practical alternative to chemical routes which yield 57-64%. PMID- 16257211 TI - Care of older people in critical care: the hidden side of the moon. PMID- 16257213 TI - Long term outcome in children with Moyamoya disease: experience with 16 patients. AB - We present our experience with 16 children with Moyamoya disease/variant diagnosed at the Royal Alexandra Hospital for Children, Westmead, Sydney, Australia in the period between January 1982 and March 2004. Thirteen of these patients had one of the different revascularisation procedures. We reviewed the modes of presentation and the long-term outcome in these children. In our series the functional outcome was related to the functional status at presentation and was not related to the type of surgical procedure or age at presentation. We believe this is the largest reported series of Moyamoya disease/variant in Australia. PMID- 16257214 TI - Protracted mumps encephalitis with good outcome. AB - There is limited published information regarding the outcome of patients with prolonged encephalitis. This report details the case of a patient with an encephalitic illness with a protracted period of coma and a favourable outcome. Extensive investigation revealed seroconversion for mumps infection. A household contact had measles, mumps, rubella (MMR) vaccination 10 days prior to his presentation. PMID- 16257215 TI - The effect of admission physiological variables on 30 day outcome after stroke. AB - INTRODUCTION: Potentially modifiable physiological variables may influence stroke prognosis but their independence from modifiable factors remains unclear. METHODS: Admission physiological measures (blood pressure, heart rate, temperature and blood glucose) and other unmodifiable factors were recorded from patients presenting within 48 hours of stroke. These variables were compared with the outcomes of death and death or dependency at 30 days in multivariate statistical models. RESULTS: In the 186 patients included in the study, age, atrial fibrillation and the National Institutes of Health Stroke Score were identified as unmodifiable factors independently associated with death and death or dependency. After adjusting for these factors, none of the physiological variables were independently associated with death, while only diastolic blood pressure (DBP) > or = 90 mmHg was associated with death or dependency at 30 days (p = 0.02). CONCLUSIONS: Except for elevated DBP, we found no independent associations between admission physiology and outcome at 30 days in an unselected stroke cohort. Future studies should look for associations in subgroups, or by analysing serial changes in physiology during the early post-stroke period. PMID- 16257216 TI - Simple partial status epilepticus in Chinese adults. AB - Simple partial status epilepticus (SPSE) is uncommon compared with generalized tonic-clonic status epilepticus. We evaluated the clinical profile and predictors of poor outcome in a group of Chinese patients with this condition. We identified 32 patients above the age of 14 years with SPSE from a large urban hospital over an eleven-year period. Factors for poor outcome, defined as death or morbidity, were analyzed. The most common underlying causes were due to cerebrovascular disease (46.9%), CNS infection (15.6%), metabolic derangement (12.5%) and tumor (12.5%). At 30 days from the onset of seizures, 13(40.5%) patients had recovered fully and seven (21.9%) had died. Poor outcome was associated with the presence of an acute symptomatic injury. PMID- 16257217 TI - Effects of citicoline on experimental spinal cord injury. AB - OBJECTIVE: To investigate the effects of citicoline on experimental spinal cord injury (SCI). BACKGROUND: Citicoline has been successfully used in clinical studies of head injury and cerebral infarction, but there is limited literature regarding its use in experimental SCI. STUDY DESIGN: Twenty adult Wistar rats were divided into four groups: sham, trauma, vehicle, and citicoline-treated. SCI was produced using a weight drop technique. Citicoline 300 mg/kg was given intraperitoneally, 5 minutes after the induction of trauma. The animals were sacrificed and 1 cm long samples of injured spinal cord were obtained at 48 hours post-SCI. Lipid peroxidation was estimated by the thiobarbituric acid test. Neurological examinations were performed using a previously described grading scale. RESULTS: Measures of lipid peroxidation and motor scores of the citicoline treated group were significantly lower than those in the other injury groups. CONCLUSIONS: Citicoline attenuated lipid peroxidation after SCI and improved the motor scores. Further investigations will be required to determine the long-term effects of this drug on spinal cord injury. PMID- 16257218 TI - Parkin expression in human skeletal muscle. AB - Parkin is known to be present in human neurons and peripheral nerves. Using an antibody against parkin protein we have now demonstrated that parkin is also expressed in the sarcoplasm and sarcolemmal region of human skeletal muscle fibres. We have also found different age-related patterns of expression with increase in intensity and organization of distribution at older ages. These findings suggest a change in the functional role of parkin in skeletal muscle with ageing and may contribute to understanding the mechanisms of muscle aging. PMID- 16257219 TI - Protein arginine methyltransferases: guardians of the Arg? AB - The recent discovery of enzymes that convert methylated arginine residues in proteins to citrulline has catapulted arginine methylation into the attention of cell-signaling researchers. Long considered a rather static post-translational modification of marginal interest, it seems that arginine methylation has now joined the group of signaling pathways that operate via pairs of antagonistic enzymes. However, many questions remain unanswered, especially concerning the removal mechanism and its implication for the physiological role of arginine methylation. I propose that, in addition to the broadly discussed function as regulator of protein activity, arginine methylation might serve a second purpose: protection of arginine residues against attack by endogenous reactive dicarbonyl agents, such as methylglyoxal, which are natural by-products of normal metabolic pathways. Inefficient detoxification of these highly cytotoxic compounds results in inactivation of proteins that is causally linked to diabetes, cancer, neurodegenerative diseases and pathophysiologies of aging. This new concept of 'arginine protection' might have far-reaching implications for the development of drugs that exploit a natural protection mechanism for medical purposes. PMID- 16257220 TI - Tendon allografts in the knee. PMID- 16257221 TI - In vitro oxidative metabolism study of (-)-rhazinilam. AB - Metabolism studies were conducted in order to investigate the reasons for the in vivo lack of activity of (-)-rhazinilam 1, an original poison of the mitotic spindle. Bioconversion by Beauveria bassiana strains, rat and human liver microsomes allowed the identification of metabolites 2, 3, and 4 oxidized in positions 3 and 5 of rhazinilam. Further experiments indicated that CYP2B6 was the main CYP responsible for the oxidation of 1 by human liver microsomes. All isolated metabolites were markedly less active than rhazinilam in vitro, which might explain its in vivo inactivity. PMID- 16257222 TI - In bacterial meningitis cortical brain damage is associated with changes in parenchymal MMP-9/TIMP-1 ratio and increased collagen type IV degradation. AB - Adverse outcome in bacterial meningitis is associated with the breakdown of the blood-brain barrier (BBB). Matrix-metalloproteinases (MMPs) facilitate this process by degradation of components of the BBB. This in turn results in acute complications of bacterial meningitis including edema formation, increased intracranial pressure and subsequent ischemia. We determined the parenchymal balance of MMP-9 and TIMP-1 (tissue inhibitor of MMP) and the structural integrity of the BBB in relation to cortical damage in an infant rat model of pneumococcal meningitis. The data demonstrate that the extent of cortical damage is significantly associated with parenchymal gelatinolytic activity and collagen type IV degradation. The increased gelatinolysis was found to be associated with a brain parenchymal imbalance of MMP-9/TIMP-1. These findings provide support to the concept that MMPs mediated disruption of the BBB contributes to the pathogenesis of bacterial meningitis and that protection of the vascular unit may have neuroprotective potential. PMID- 16257223 TI - Transforming growth factor beta2 haploinsufficient mice develop age-related nigrostriatal dopamine deficits. AB - The transforming growth factor-betas (TGF-betas) regulate the induction of dopaminergic neurons and are elevated in the CSF of Parkinson's patients. We report here that mice with TGF-beta2 haploinsufficiency (TGF-beta2+/-) have subclinical defects in the dopaminergic neurons of their substantia nigra pars compacta. At 6 weeks of age, the TGF-beta2+/- mice had 12% fewer dopaminergic neurons than wild-type littermates. No additional loss of neurons occurred during the next 5 months, although striatal dopamine declined to 70% of normal. The level of 3,4-dihydroxphenylacetic acid was normal in the TGF-beta2+/- mice, indicating that a compensatory mechanism maintains dopamine stimulation of their striatum. The TGF-beta2+/- mice had normal sensitivity to the neurotoxin 1-methyl 4-phenyl-1,2,3,6-tetrahydropyridine, despite having reduced levels of monoamine oxidase-B. These results raise the possibility that people with naturally low levels of TGF-beta2 may have less functional reserve in their nigrostriatal pathway, causing them to be at increased risk of developing Parkinson disease. PMID- 16257224 TI - Alzheimer's disease: mRNA expression profiles of multiple patients show alterations of genes involved with calcium signaling. AB - We combined global and high-resolution strategies to find genes with altered mRNA expression levels in one of the largest collection of brain autopsies from Alzheimer's patients and controls ever studied. Our global analysis involved microarray hybridizations of large pools of samples obtained from 114 individuals, using two independent sets of microarrays. Ten genes selected from the microarray experiments were quantified on each individual separately using real-time RT-PCR. This high-resolution analysis accounted for systematic differences in age, postmortem interval, brain pH, and reference gene expression, and it estimated the effect of disease on mRNA levels, on top of the effect of all other variables. Differential expression was confirmed for eight out of ten genes. Among them, Type B inositol 1,4,5-trisphosphate 3-kinase (ITPKB), and regulator of G protein signaling 4 (RGS4) showed highly altered expression levels in patients (P values < 0.0001). Our results point towards increased inositol triphospate (IP3)-mediated calcium signaling in Alzheimer's disease. PMID- 16257225 TI - The generation of a conditional Fmr1 knock out mouse model to study Fmrp function in vivo. AB - The FMR1 gene, mutated in Fragile X syndrome patients, has been modeled in mice with a neomycin cassette inserted in exon 5 of the mouse Fmr1 gene creating an Fmr1 knockout (Fmr1 KO) allele. This results in animals lacking Fmr1 protein (Fmrp) expression in all tissues. We have created a new, more versatile Fmr1 in vivo KO model (Fmr1 KO2) and generated conditional Fmr1 KO (CKO) mice by flanking the promoter and first exon of Fmr1 with lox P sites. This enables us to create a null allele in specific cell types and at specific time points by crossing Fmr1 CKO mice with tissue specific or inducible cre-recombinase expressing mice. The new Fmr1 KO2 line does not express any Fmrp and also lacks detectable Fmr1 transcripts. Crossing the Fmr1 CKO line with a Purkinje cell-specific cre recombinase expresser produces mice that are null for Fmr1 in Purkinje neurons but wild type in all other cell types. PMID- 16257226 TI - A novel role of circadian transcription factor DBP in hippocampal plasticity. AB - In neurons, a variety of extracellular stimuli are capable of inducing transcriptional events that underlie complex processes ranging from learning to disease. The mechanisms linking these long-lasting cellular modifications to behavior remain to be established. Here, we show by microarray analysis that hippocampal activation of glucagon-like peptide-1 receptor (GLP-1R), which is associated with improved learning and neuroprotection, results in suppression of the transcription factor DBP (albumin D-site-binding protein). Recombinant adeno associated virus (rAAV) based gene expression of DBP in the hippocampus of adult rats caused upregulation of mRNAs encoding constituents of the molecular clock, and the DBP target gene, pyridoxal kinase. Behaviorally, DBP over expression inhibited spatial learning but not memory, and enhanced susceptibility to kainate induced seizures. This phenotype was paralleled by the activation of MAP kinase in dendritic regions of hippocampal neurons in vivo. These data suggest that DBP may represent an important transcriptional link between GLP-1R activation and neuroplasticity in the hippocampus. PMID- 16257227 TI - Health data linkage conserves privacy in a research-rich environment. PMID- 16257228 TI - Racial disparities in elevated prenatal depressive symptoms among black and white women in eastern north Carolina. AB - PURPOSE: Black women have an increased risk for preterm birth compared with white women, and prior research indicated that maternal prenatal depressive symptoms are associated with increased risk for preterm outcomes among black women. Race related differences in prenatal depression could be of etiologic significance in understanding racial disparities in preterm birth. Our study focused on Center for Epidemiologic Studies' Depression Scale (CES-D) scores of pregnant black and white women. METHODS: Women were administered the CES-D at the time of their first visit to hospital-based prenatal clinics. Two cutoff scores for the CES-D were used: 16 or higher, which indicates "significant" depressive symptoms, and 23 or higher, which indicates major depressive disorder. RESULTS: For the sample of 1163 women, mean CES-D scores were significantly higher among black (17.4) than white (13.7) women. Of black women, 49% had CES-D scores higher than 15 compared with 33.5% of white women. Also, 27.5% of black women had scores higher than 22 compared with 16% of white women. After adjustment for maternal age, marital status, and education, odds ratios for race for both CES-D cutoff scores were approximately 1.5. CONCLUSIONS: Results of this study indicate that black women have greater rates of prenatal depression than white women. PMID- 16257229 TI - Pattern of alcohol consumption during pregnancy and risk for low birth weight. AB - PURPOSE: The effects of moderate alcohol drinking on low birth weight are not clear, and conflicting results have been reported. We assessed the influence of different patterns of alcohol drinking during pregnancy on low birth weight. METHODS: A case-control study was carried out at the University of Cantabria Hospital, Spain, from 1998 to 2002. Cases (n = 552) were mothers delivering a single newborn weighing less than 2500 g. Controls (n = 1451) were selected from a random sample of all delivering women. Information was obtained from personal interview, clinical charts, and prenatal care records. Odds ratios (ORs) and 95% confidence intervals (CIs) were estimated. Adjustment for potential confounders was made by means of logistic regression. RESULTS: Alcohol consumption of less than 6 g/day decreased the risk for low birth weight (adjusted OR = 0.64; 95% CI, 0.46-0.88). A similar result was obtained for moderate drinkers (<12 g/day) on weekends only. The opposite relationship was observed between alcohol consumption on weekdays of 12 g/day or greater (adjusted OR = 2.67; 95% CI, 1.39-5.12), not observed in those drinking on weekends only. The interaction between alcohol consumption and tobacco smoking was analyzed. Weekday drinkers of 12 g/day or greater showed an increased risk in smokers. Alcohol consumption on weekends only in nonsmokers was inversely related. The influence of alcohol was greater for small-for-gestational-age (SGA) than non-SGA babies. CONCLUSIONS: Alcohol consumption of 12 g/day or greater increased the risk for low birth weight, whereas lower consumption during weekends showed the opposite effect (mainly in nonsmokers). PMID- 16257230 TI - Is the factor structure of the metabolic syndrome comparable between men and women and across three ethnic groups: the Miami Community Health Study. AB - PURPOSE: The metabolic syndrome (MS) is conceived as a cluster of disorders that increase risk for coronary heart disease and type 2 diabetes. Studies examining its structure primarily have used an exploratory factor analytic technique, but yielded discrepant results. There also is a lack of research that investigates whether the clustering pattern is similar across sex and ethnic groups. This study uses confirmatory factor analysis to evaluate the factor structure of the MS and examines its similarity between men and women and across three ethnic groups (Caucasian, African, and Cuban Americans). METHOD: A hierarchical four factor model with an overarching MS factor uniting insulin resistance, obesity, lipid, and blood pressure factors was tested with 517 individuals from the Miami Community Health Study. RESULTS: Findings show that the proposed structure was well supported (comparative fit index=0.97) and similar between men and women and across ethnic groups. The MS was represented strongly by insulin resistance, followed by obesity, lipid factors, and, to a lesser extent, a blood pressure factor. CONCLUSIONS: This study provides empirical support for identifying and diagnosing the MS by its component factors in a diverse population. PMID- 16257231 TI - Spousal correlations for lifestyle factors and selected diseases in Chinese couples. AB - PURPOSE: Spouses usually are genetically unrelated and share a common living environment. Thus, concordance of diseases in spouses reflects mainly environmental etiologic contributors. The purpose of this study is to investigate spousal associations for selected lifestyle characteristics and common medical conditions. METHODS: Baseline information from 66,130 married couples participating in the Shanghai Women's Health Study was used in this analysis. Husband-wife associations were evaluated by means of logistic regression, using women's lifestyle and medical conditions as dependent variables. Adjustments were made for women's age, education, occupation, and family income in all models. RESULTS: Women were more than twice as likely to be current or former smokers; be regular consumers of alcohol, tea, and ginseng; and exercise regularly if their husbands had the same habit. A statistically significant husband-wife disease association was found for tuberculosis, chronic bronchitis, asthma, chronic gastritis, chronic hepatitis, ulcerative colitis, cholelithiasis, high blood pressure, coronary heart disease, and stroke. CONCLUSIONS: Spouses share common lifestyle habits and health risks. This study supports the hypothesis that the shared marital environment may contribute to similarities in lifestyle and morbidity in spouses and provides a basis for health promotion and prevention strategies that target the spouses of patients. PMID- 16257232 TI - Systematic review of the influence of childhood socioeconomic circumstances on risk for cardiovascular disease in adulthood. AB - PURPOSE: Adverse socioeconomic circumstances in childhood may confer a greater risk for adult cardiovascular disease (CVD). The purpose of this review is to systematically evaluate evidence for an association between socioeconomic circumstances during childhood and specific CVD subtypes, independent of adult socioeconomic conditions. METHODS: We systematically retrieved individual-level studies of morbidity and mortality from CVD and specific CVD subtypes linked to early life influences, including coronary heart disease (CHD), ischemic and hemorrhagic stroke, peripheral vascular disease, markers of atherosclerosis (carotid intima-media thickness and stenosis), and rheumatic heart disease. Indicators of socioeconomic position in childhood varied, although most studies relied on father's occupation. RESULTS: We located 40 studies (24 prospective, 11 case-control, and 5 cross-sectional) reported in 50 publications. Thirty-one studies (19 prospective, 7 case-control, and all 5 cross-sectional) found a robust inverse association between childhood circumstances and CVD risk, although findings sometimes varied among specific outcomes, socioeconomic measures, and sex. Case-control studies reported mixed results. The association was stronger for stroke and, in particular, hemorrhagic stroke, than for CHD. Childhood socioeconomic conditions remained important predictors of CVD, even in younger cohorts. CONCLUSION: Childhood and adulthood socioeconomic circumstances are important determinants of CVD risk. The specific contribution of childhood and adulthood characteristics varies across different CVD subtypes. Disease-specific mechanisms are likely to explain the childhood origins of these adult health inequalities. PMID- 16257233 TI - Immunostimulatory nature of beta-glucans and baker's yeast in gnotobiotic Artemia challenge tests. AB - The use of new preventive approaches such as immunostimulants to reduce stress and mortalities, to maintain good health of cultured organisms and to stimulate the non-specific defence mechanism, is becoming increasingly important in aquaculture. Yet detailed analysis reveals that in most experiments the validity of some conclusions with respect to the benefit of immunostimulation is still doubtful, especially in invertebrates. The use of standardized trials under controlled rearing conditions, complemented with fundamental research on defence mechanisms can provide unequivocal evidence for the beneficial effects of immunostimulants in reducing invertebrate susceptibility to diseases or infections. This study investigated the use of small amounts of baker's yeast Saccharomyces cerevisiae and glucan particles (obtained from baker's yeast) in gnotobiotic Artemia to overcome the pathogenicity of two organisms: Vibrio campbellii and V proteolyticus. Artemia supplemented with small quantities of a yeast strain presenting higher concentrations of beta-glucans or with glucan particles seemed to completely resist the detrimental effects of both pathogens. The higher amount and/or availability of beta-glucans in that yeast might play an essential role in such protection, as most probably glucans stimulate the immune response of the nauplii. PMID- 16257234 TI - Varieties of self-systems worth having. PMID- 16257235 TI - On rotational invariance in adaptive spatial filtering of fMRI data. AB - Canonical correlation analysis (CCA) has previously been shown to work well for detecting neural activity in fMRI data. The reason is that CCA enables simultaneous temporal modeling and adaptive spatial filtering of the data. This article introduces a novel method for adaptive anisotropic filtering using the CCA framework and compares it to a previously proposed method. Isotropic adaptive filtering, which is only able to form isotropic filters of different sizes, is also presented and evaluated. It is shown that a new feature of the proposed method is invariance to the orientation of activated regions, and that the detection performance is superior to both that of the previous method and to isotropic filtering. PMID- 16257236 TI - Investigating the post-stimulus undershoot of the BOLD signal--a simultaneous fMRI and fNIRS study. AB - Measuring the hemodynamic response with functional magnetic resonance imaging (fMRI) together with functional near-infrared spectroscopy (fNIRS) may overcome limitations of single-method approaches. Accordingly, we measured the event related hemodynamic response with both imaging methods simultaneously in young subjects during visual stimulation. An intertrial interval of 60 s was chosen to include the prolonged post-stimulus undershoot of the blood oxygenation level dependent (BOLD) signal. During visual stimulation, the BOLD signal, oxy-, and total hemoglobin (Hb) increased, whereas deoxy-Hb decreased. The post-stimulus period was characterized by an undershoot of the BOLD signal, oxy-Hb, and an overshoot of deoxy-Hb. Total Hb as measured by fNIRS returned to baseline immediately after the end of stimulation. Results suggest that the post-stimulus events as measured by fNIRS are dominated by a prolonged high-level oxygen consumption in the microvasculature. The contribution of a delayed return of blood volume to the BOLD post-stimulus undershoot in post-capillary veins as suggested by the Balloon and Windkessel models remains ambiguous. Temporal changes in the BOLD signal were highly correlated with deoxy-Hb, with lower correlation values for oxy- and total Hb. Furthermore, data show that fNIRS covers the outer 1 cm of the brain cortex. These results were confirmed by simultaneous fMRI/fNIRS measurements during rest. In conclusion, multimodal imaging approaches may contribute to the understanding of neurovascular coupling. PMID- 16257237 TI - Spatial specificity of the enhanced dip inherently induced by prolonged oxygen consumption in cat visual cortex: implication for columnar resolution functional MRI. AB - Since changes in oxygen consumption induced by active neurons are specific to cortical columns, the small and transient "dip" of deoxyhemoglobin signal, which indicates an increase in oxygen consumption, has been of great interest. In this study, we succeeded in enhancing and sustaining the dip in the deoxyhemoglobin weighted 620-nm intrinsic optical imaging signals from a 10-s orientation selective stimulation in cat visual cortex by reducing arterial blood pressure with sodium nitroprusside (a vasodilator) to mitigate the contribution of stimulus-induced blood supply. During this condition, intact spiking activity and a significant reduction of stimulus-induced blood volume changes (570-nm intrinsic signals) were confirmed. The deoxyhemoglobin signal from the prolonged dip was highly localized to iso-orientation domains only during the initial approximately 2 s; the signal specificity weakened over time although the domains were still resolvable after 2 s. The most plausible explanation for this time dependent spatial specificity is that deoxyhemoglobin induced by oxygen consumption drains from active sites, where spiking activity occurs, to spatially non-specific downstream vessels over time. Our results suggest that the draining effect of pial and intracortical veins in dHb-based imaging techniques, such as blood oxygenation-level dependent (BOLD) functional MRI, is intrinsically unavoidable and reduces its spatial specificity of dHb signal regardless of whether the stimulus-induced blood supply is spatially specific. PMID- 16257238 TI - Optimization of dynamic measurement of receptor kinetics by wavelet denoising. AB - The most important technical limitation affecting dynamic measurements with PET is low signal-to-noise ratio (SNR). Several reports have suggested that wavelet processing of receptor kinetic data in the human brain can improve the SNR of parametric images of binding potential (BP). However, it is difficult to fully assess these reports because objective standards have not been developed to measure the tradeoff between accuracy (e.g. degradation of resolution) and precision. This paper employs a realistic simulation method that includes all major elements affecting image formation. The simulation was used to derive an ensemble of dynamic PET ligand (11C-raclopride) experiments that was subjected to wavelet processing. A method for optimizing wavelet denoising is presented and used to analyze the simulated experiments. Using optimized wavelet denoising, SNR of the four-dimensional PET data increased by about a factor of two and SNR of three-dimensional BP maps increased by about a factor of 1.5. Analysis of the difference between the processed and unprocessed means for the 4D concentration data showed that more than 80% of voxels in the ensemble mean of the wavelet processed data deviated by less than 3%. These results show that a 1.5x increase in SNR can be achieved with little degradation of resolution. This corresponds to injecting about twice the radioactivity, a maneuver that is not possible in human studies without saturating the PET camera and/or exposing the subject to more than permitted radioactivity. PMID- 16257239 TI - Amygdala activation when one is the target of deceit: did he lie to you or to someone else? AB - The ability to figure out whether a person is being honest or deceitful is an important part of social competence. Reactions to deceit may however differ depending on whether one is being deceived oneself or observes a deceitful exchange between others. In the present study, we investigated whether personal involvement influenced the neural responses associated with the detection of deceit. Subjects watched videos of actors lifting a box and judged whether the actors had been misled about the real weight of the box. Personal involvement was manipulated by having the participants themselves among the actors. The critical finding was that there was activity in amygdala and fusiform gyrus only for the condition in which participants observed themselves being deceived. In contrast, the superior temporal sulcus and anterior cingulate cortex were activated irrespective of whether the participants detected that the experimenter had deceived themselves or another. These four brain areas are all interconnected and are part of the discrete neural system subserving social cognition. Our results provide direct evidence, using judgments of deceit in a social context, that the crucial factor for amygdala activation is the involvement of the subjects because they are the target of the deceit. We interpret the activation of the amygdala in this situation as reflecting the greater affective reaction when one is deceived oneself. Our results suggest that when one is personally involved, deceit is taken as a potential threat. PMID- 16257240 TI - Probabilistic 3D MRI atlas of the human cerebellar dentate/interposed nuclei. AB - In a previous study, a three-dimensional (3D) MRI atlas of the human cerebellar nuclei was introduced based on findings in one healthy human subject [Dimitrova, A., Weber, J., Redies, C., Kindsvater, K., Maschke, M., Kolb, F.P., Forsting, M., Diener, H.C., Timmann, D., 2002. MRI atlas of the human cerebellar nuclei. NeuroImage 17, 240-255]. The present MRI investigation was designed to study variability of the anatomy of the dentate/interposed nuclei in a larger group of healthy subjects. Similar to our previous study, iron-induced susceptibility artifacts were used to visualize the cerebellar nuclei as hypointensities on MR images. Data of 63 healthy subjects (27 female, 36 male; mean age 45.3+/-13.4 years, age range 22--71 years) were included. A 3D axial volume of the cerebellum was acquired using a T2*-weighted FLASH sequence on a Siemens Sonata 1.5 T MR scanner. Each volume was registered, re-sampled to 1.00 x 1.00 x 1.00 mm(3) voxel size and spatially normalized into a standard proportional stereotaxic space using SPM99. Dentate/interposed nuclei were traced on axial images and saved as regions of interest using MRIcro-software by two independent examiners. A probabilistic 3D MRI atlas of the cerebellar dentate/interposed nuclei is presented based on findings in all subjects. PMID- 16257241 TI - Local origin and diversification among Lampornis hummingbirds: a Mesoamerican taxon. AB - The huge biodiversity found in Mesoamerica is often explained by its geographic situation as a natural bridge between two large biogeographic regions. Often overlooked, however, are the high levels of speciation and diversification in the area. Here we assess the phylogenetic relationships within a Mesoamerican group of hummingbirds (Lampornis). We sequenced both mtDNA (1,143 bp of cyt b and 727 bp of ND5) and nuclear genes (505 bp of AK-5 intron and 567 bp of c-mos) for each of the seven recognised species and outgroups. We find two or three clades of similar age within this genus: L. clemenciae and L. amethystinus (singly or as each other's sister taxa) and a Central American clade. This Central-American clade presents a clear bipartition between northern (L. viridipallens and L. sybillae) and southern Mesoamerica, which is shared with many other Mesoamerican organisms. Our analyses suggest that L. hemileucus does not belong in the genus Lampornis. While we refrain to apply a time-scale to our data because of the lack of an appropriate calibration, our results indicate that the genus Lampornis predates the uprising of the Panama land-bridge, and that diversification among the isthmian species (L. castaneoventris and L. calolaema) is a very recent event. Our results strongly suggest a local Mesoamerican origin for this genus. PMID- 16257242 TI - A contentious issue finds some clarity: on the independent and complementary roles of aggrecanase activity and MMP activity in human joint aggrecanolysis. AB - Our understanding of aggrecanolysis in the human joint has recently been clarified by detailed analysis of naturally occurring intermediates in cartilage and synovial fluids. The most studied aspect has been the proteolysis of the interglobular domain (IGD) of aggrecan with release of the glycosaminoglycan (GAG)-attachment regions, because this appears to be most destructive to tissue function. In this Editorial review, a working model is presented which supports the view that one or more aggrecanases (ADAMTS 1, 4, 5, 8, 9, 15) are responsible for cleavage of the IGD with destructive loss of tissue GAG. In contrast, one or more metalloproteinases (MMPs) (MMP 1, 2, 3, 7, 8, 9, 10, 13, 14, 19, 20) are responsible for cleavage of the IGD (at Asn360-Phe361) within a separate pool of aggrecan, which does not bear GAG, because it has previously been C-terminally truncated in a separate slow turnover process. These findings, along with recent gene deletion studies in mice, suggest that ADAMTS-mediated aggrecanolysis is destructive to cartilage function whereas MMP-mediated aggrecanolysis may actually be beneficial. PMID- 16257243 TI - Chondrogenic differentiation and functional maturation of bovine mesenchymal stem cells in long-term agarose culture. AB - BACKGROUND: The developmental history of the chondrocyte results in a cell whose biosynthetic activities are optimized to maintain the concentration and organization of a mechanically functional cartilaginous extracellular matrix. While useful for cartilage tissue engineering studies, the limited supply of healthy autologous chondrocytes may preclude their clinical use. Consequently, multipotential mesenchymal stem cells (MSCs) have been proposed as an alternative cell source. OBJECTIVE: While MSCs undergo chondrogenesis, few studies have assessed the mechanical integrity of their forming matrix. Furthermore, efficiency of matrix formation must be determined in comparison to healthy chondrocytes from the same donor. Given the scarcity of healthy human tissue, this study determined the feasibility of isolating bovine chondrocytes and MSCs, and examined their long-term maturation in three-dimensional agarose culture. EXPERIMENTAL DESIGN: Bovine MSCs were seeded in agarose and induced to undergo chondrogenesis. Mechanical and biochemical properties of MSC-laden constructs were monitored over a 10-week period and compared to those of chondrocytes derived from the same group of animals maintained similarly. RESULTS: Our results show that while chondrogenesis does occur in MSC-laden hydrogels, the amount of the forming matrix and measures of its mechanical properties are lower than that produced by chondrocytes under the same conditions. Furthermore, some important properties, particularly glycosaminoglycan content and equilibrium modulus, plateau with time in MSC-laden constructs, suggesting that diminished capacity is not the result of delayed differentiation. CONCLUSIONS: These findings suggest that while MSCs do generate constructs with substantial cartilaginous properties, further optimization must be done to achieve levels similar to those produced by chondrocytes. PMID- 16257244 TI - Autosomal dominant hereditary hemochromatosis associated with two novel Ferroportin 1 mutations in Spain. AB - Hereditary hemochromatosis is a common disorder of iron metabolism most frequently associated with mutations in the HFE gene. Hereditary hemochromatosis may be caused by other genetic mutations including those in the SLC40A1 gene. This report describes the clinical and laboratory findings of two Spanish families with autosomal dominant iron overload associated with previously unrecognized Ferroportin 1 mutations (p.R88T and p.I180T). The phenotype of iron overload in the patients carrying these mutations could correspond to the group of clinical mutations that lose their iron export function. PMID- 16257245 TI - Bach flower remedies used for attention deficit hyperactivity disorder in children--a prospective double blind controlled study. AB - OBJECTIVE: To evaluate the effectiveness of Bach flower remedies in the treatment of children with attention deficit hyperactivity disorder (ADHD), in a double blind prospective controlled study. METHODS: Fourty Children with ADHD, aged 7-11 years, diagnosed according to the DSM criteria, were randomised to Bach flower remedies or placebo treatments for a period of 3 months. Children's performance was evaluated by the teacher before commencement of treatment and subsequently each month during the study period. RESULTS: Bach flower remedies have no statistically significant effect when compared to placebo in the treatment of children with ADHD. There was a significant correlation between treatment duration's and improvement of performance, with no difference between the treatment group compared to the placebo. CONCLUSIONS: There is no statistically significant difference between the effects of Bach flower remedies compared with placebo in the treatment of children with ADHD. PMID- 16257246 TI - Prevalence of pyridoxine dependent seizures in south Indian children with early onset intractable epilepsy: A hospital based prospective study. AB - OBJECTIVES: Determine the prevalence of pyridoxine dependent seizures in children less than 16 years of age attending a teaching hospital in south India with early onset (before 3 years) intractable epilepsy of unknown aetiology, using the criteria proposed by Baxter. METHOD: A cohort of 81 children, fulfilling the above criteria, was given 15 mg/kg/day of oral pyridoxine for 7 days. Non responders were given a further 7-day trial of 30 mg/kg/day. Diagnosis of pyridoxine dependent seizures was made according to the criteria proposed by Baxter. RESULT: Six children (7.4%; four boys and two girls) were identified as definite cases and were continued on B6, without recurrence of seizures. Median age of seizure onset and diagnosis were 2.5 months and 2 year 9 months, respectively. No significant complications were observed with B6 therapy. CONCLUSION: Chance of identifying PDS is not low in a subpopulation of children with intractable early onset cryptogenic epilepsy using the criteria proposed by Baxter, which warrants a therapeutic trial with B6 in them. PMID- 16257247 TI - Novel properties of malarial S-adenosylmethionine decarboxylase as revealed by structural modelling. AB - In the malaria parasite, the two main regulatory activities of polyamine biosynthesis, ornithine decarboxylase (ODC) and S-adenosylmethionine decarboxylase (AdoMetDC) occur in a single bifunctional protein. The AdoMetDC domain was modeled using the human and potato X-ray crystal structures as templates. Three parasite-specific inserts and the core active site region was identified using a structure-based alignment approach. The domain was modeled without the two largest inserts, to give a root mean square deviation of 1.85 angstroms from the human template. Contact with the rest of the bifunctional complex is predicted to occur on one face of the Plasmodium falciparum AdoMetDC (PfAdoMetDC) domain. In the active site there are four substitutions compared to the human template. One of these substitutions may be responsible for the lack of inhibition by Tris, compared to mammalian AdoMetDC. The model also provides an explanation for the lack of putrescine stimulation in PfAdoMetDC compared to mammalian AdoMetDC. A network of residues that connects the putrescine-binding site with the active site in human AdoMetDC is conserved in the malarial and plant cognates. Internal basic residues are found to assume the role of putrescine, based on the model and site-directed mutagenesis: Arg11 is absolutely required for normal activity, while disrupting Lys15 and Lys215 each cause 50% inhibition of AdoMetDC activity. These novel features of malarial AdoMetDC suggest possibilities for the discovery of parasite-specific inhibitors. PMID- 16257248 TI - Arousal and re-feeding rapidly restores digestive tract morphology following aestivation in green-striped burrowing frogs. AB - During aestivation, the gut of the green-striped burrowing frog, Cyclorana alboguttata undergoes significant morphological down-regulation. Despite the potential impact such changes might have on the re-feeding efficiency of these animals following aestivation, they appear to be as efficient at digesting their first meals as active, non-aestivating animals. Such efficiency might come about by the rapid restoration of intestinal morphology with both arousal from aestivation and the initial stages of re-feeding. Consequently, this study sought to determine what morphological changes to the intestine accompany arousal and re feeding following 3 months of aestivation. Arousal from aestivation alone had a marked impact on many morphological parameters, including small and large intestine masses, small intestinal length, LF heights, enterocyte cross-sectional area and microvilli height and density. In addition, the onset of re-feeding was correlated with an immediate reversal of many morphological parameters affected by 3 months of aestivation. Those parameters that had not returned to control levels within 36 h of feeding generally had returned to control values by the completion of digestion (i.e. defecation of the meal). Re-feeding was also associated with several changes in enterocyte morphology including the incorporation in intracytoplasmic lipid droplets and the return of enterocyte nuclear material to the 'active' euchromatin state. In conclusion, morphological changes to the gut of aestivating frogs which occur during aestivation are transitory and rapidly reversible with both arousal from aestivation and re feeding. The proximate causes behind these transitions and their functional significance are discussed. PMID- 16257249 TI - The selenium intake of the female chicken influences the selenium status of her progeny. AB - The primary purpose of this study is to determine the extent to which the effects of dietary supplementation of the female chicken with selenium (Se) continue into the next generation. An additional aim is to compare the relative effectiveness of pre-hatch (from the hen's diet) with that of post-hatch (from the progeny's diet) supplementation with Se on the Se status of the chick during the first 4 weeks of post-hatch life. Hens were maintained on control or Se-supplemented diets, respectively containing 0.027 and 0.419 mug Se/g of feed. The high-Se diet elevated the Se content of the hens' eggs by 7.1-fold. At hatch, the concentrations of Se in the liver, breast muscle and whole blood of the chicks originating from the high-Se parents were, respectively, 5.4-, 4.3- and 7.7-fold higher than the values in the chicks of the low-Se parents. When the offspring from the two parental groups were both maintained on the low-Se progeny diet, the tissue Se concentrations in chicks originating from the high-Se hens remained significantly higher for 3-4 weeks after hatching, compared with the values in chicks from the low-Se hens. Similarly, tissue glutathione peroxidase activity remained significantly higher in chicks from the high-Se hens for 2-4 weeks post hatch. Thus, the effects of maternal Se supplementation persist in the progeny for several weeks after hatching. However, when chicks hatching from low-Se eggs were placed on a high Se diet, their tissue Se concentrations at 7 days of age were markedly higher than the values in chicks from high-Se eggs placed on the low-Se diet. PMID- 16257250 TI - Molecular characterization and expression analysis of vitellogenin in the marine crab Portunus trituberculatus. AB - As a precursor of the major yolk protein vitellin (Vn), vitellogenin (Vg) has been studied in crustacean for decades, mainly in shrimp. However, little is known about the Vg molecule in crabs. In the present study, we report a cDNA encoding Vg in the marine crab Portunus trituberculatus, an important commercial species in aquaculture. The cDNA sequence is 7,846 bp in length, containing a 7,680 bp open reading frame, that encodes 2,560 amino acid residues. The deduced amino acid sequence showed 32-75% identity with other known crustacean Vgs. Seven potential cleavage sites (RXXR) were identified in the deduced Vg precursor. SDS PAGE and Western blotting analysis demonstrated that Vns in the ovary of the mature crab consisted of three major polypeptides (102, 100 and 85 kDa). Northern blotting analysis revealed that Vg was mainly expressed in hepatopancreas in vitellogensis females. A Vg transcript was also identified in the ovary of the crab by reverse transcription-polymerase chain reaction analysis. PMID- 16257251 TI - Transcriptional responses in mouse lungs induced by vaccination with Mycobacterium bovis BCG and infection with Mycobacterium tuberculosis. AB - Transcriptome analyses enable the assessment of signature alterations in whole tissues and organs undergoing pathological processes. We analyzed gene expression profiles of lungs from mice infected with Mycobacterium tuberculosis or vaccinated with Mycobacterium bovis bacille Calmette-Guerin (BCG). We compared high-dose systemic and low-dose aerosol M. tuberculosis infections as well as systemic BCG vaccination. Expression profiles in lungs were analyzed at day (d) 1 and d 30 post infection / vaccination using a custom tailored 'in situ' synthesized 60-mer oligonucleotide microarray with focus on immunologically relevant genes. At d 1, a small number of genes were differentially regulated, whereas at d 30, a discrete expression pattern was identified in the lung. Differential gene expression profiles between M. tuberculosis infection and BCG vaccination indicate differences in naturally and vaccine induced pulmonary immune responses. The shared signature of systemic and aerosol M. tuberculosis infection revealed dominance of genes related to or controlled by interferon gamma (IFN-gamma). We assume that differential gene expression profiles after M. tuberculosis infection are strongly influenced by differences in cellular composition of the lung due to migration of immune cells, primarily neutrophils, basophils, eosinophils and monocytes to the site of infection. PMID- 16257252 TI - A new method for measurement of bone deformation by echo tracking. AB - No method has been available to noninvasively detect bone deformation or strain under loading in vivo. We focused on ultrasonic measurement of the displacement at a certain point on a bone using the echo-tracking method (ET). To develop a method that can noninvasively detect bone deformation in vivo, a preliminary investigation was performed. We investigated the accuracy of measuring displacement with our echo tracking system by using a flat metal panel and found that the method could measure displacement with a precision of a few microns. A three-point bending test of a porcine tibia with both ends fully constrained was performed to measure bone surface displacement, and simultaneous measurement of the surface strain was done using two strain gauges. The correlation between the displacement measured by ET and the strain gauge readings was completely linear (r=0.999), showing that the method could precisely detect bone deformation. The loads versus displacement curves obtained with cyclic loading were typical hysteresis loops that showed viscoelastic properties of the measured bone. We also improved a multi-ET system capable of simultaneously tracking multiple points to detect deformation of the bone surface. Measurement by this echo tracking system was also compared with strain gauge readings during a three point bending test with both ends of the tibia supported. The linearity of both methods was very high (r=0.998). Our ET method might have considerable potential for noninvasive measurement of bone viscoelasticity and plasticity. PMID- 16257253 TI - A comparative FEA of the debonding process in different concepts of cemented hip implants. AB - Debonding of the stem-cement interface and damage accumulation in the cement mantle are basic events that contribute to the long-term failure of cemented hip reconstructions. In this work, a numerical study with these two processes coupled is presented. On the one hand, debonding of the stem-cement interface was simulated by means of a cohesive surface theory that was implemented into an interface finite element. This interface model includes a tensile-shear behavior law, the fatigue failure of the interface, and the friction evolution between both surfaces. On the other hand, damage accumulation in the cement was formulated through the theory of continuum damage mechanics, considering cement damage due to tension, creep under compression, crack closure effects, non-linear damage accumulation and cement residual stresses appearing during polymerisation. This methodology was applied to simulate and compare the degradation process of the cement and stem-cement interface in four different concepts of design: Exeter, Charnley, Elite Plus and ABG II stems. As the actual mechanical properties of the surface of each specific prosthesis are not known, we assumed the same for all of them, distinguishing between polished and matt surfaces. With this assumption, the predicted results showed that the debonding process is very different for each implant depending on the stem geometry. Lower cement deterioration was obtained for the Exeter and ABG II stems, while the lowest stem cement interface debonding was produced in the Exeter and the Elite Plus stems. PMID- 16257254 TI - Disease-related and drug-induced changes in dopamine transporter expression might undermine the reliability of imaging studies of disease progression in Parkinson's disease. AB - Parkinson's disease (PD) is a progressive neurodegenerative disorder. Standard therapeutic interventions are aimed at replenishment of empty dopamine stores with levodopa or substitution with dopamine receptor agonists. However, in the long term this symptomatic therapy fails. Currently, various neuroprotective agents are being developed, with the intention to slow down the degeneration of dopaminergic neurons. In this context, the early identification of persons at risk to develop the disease as well as the assessment of the effectiveness of putative neuroprotective agents, are critical issues. Dopamine transporter (DAT) scintigraphy with single photon emission computed tomography (SPECT) has been used to assess the dopaminergic function in PD. Initial studies with several radioligands show significant loss of DAT binding in PD patients as compared to controls. In this paper we review the evidence on the utility of DAT imaging with SPECT in early PD detection as well as in monitoring neurprotection. PMID- 16257255 TI - Nicotine stimulates adhesion molecular expression via calcium influx and mitogen activated protein kinases in human endothelial cells. AB - To evaluate the effect of nicotine on endothelium dysfunction and development of vascular diseases, we investigated the influence on adhesion molecular expression mediated by nicotine and the mechanism of this effect in human umbilical vein endothelial cells (HUVECs). The result showed that nicotine could induce surface/soluble vascular cell adhesion molecule (VCAM-1) and endothelial selectin (E-selectin) expression in a time-response decline manner and the peak appeared at 15 min. This action could be mediated by mitogen-activated protein kinase/extracellular signal regulated kinase 1/2 (MAPK/ERK1/2) and MAPK/p38 because their activation could be distinctly blocked by MAPK inhibitors, PD098059 or SB203580. Mecamylamine (non-selective nicotinic receptor inhibitor), alpha bungarotoxin (alpha7 nicotinic receptor inhibitor) could block Ca2+ accumulation, and then, prevented the phosphorylation on ERK1/2 and p38. They also inhibited the surface/soluble VCAM-1, E-selectin production of HUVECs modulated by nicotine. Therefore, we concluded that: (i) nicotine obviously up-regulates VCAM 1 and E-selectin expression at 15 min in HUVECs, (ii) nicotine activates HUVECs triggered by the ERK1/2 and p38 phosphorylation with an involvement of intracellular calcium mobilization chiefly mediated by alpha7 nicotinic receptor, (iii) intracellular Ca2+ activates a sequential pathway from alpha7 nicotinic receptor to the phosphorylation of ERK1/2, p38. These elucidate that nicotine activates HUVECs through fast signal transduction pathway and arguments their capacity of adhesion molecular production. Further more nicotine may contribute its influence to the progression of vascular disease such as atherosclerotic lesion. PMID- 16257256 TI - Transforming growth factor-beta pathway: role in pancreas development and pancreatic disease. AB - The pancreas is a complex exocrine and endocrine gland that controls many homeostatic functions. The exocrine pancreas produces and secretes digestive enzymes, whereas, the endocrine pancreas produces four distinct hormones, chief among them being the glucose regulating hormone-insulin. Diabetes, pancreatitis and pancreatic cancer are some of the main afflictions that result from pancreas dysfunction. Transforming growth factor-beta (TGF-beta) proteins are central regulators of pancreas cell function, and have key roles in pancreas development and pancreatic disease. Since expression levels and kinase activities of components of TGF-beta signaling are aberrantly altered in diseases of the pancreas, modulating the activity of TGF-beta provides a unique and rational opportunity for therapeutic intervention. Although TGF-beta still remains elusive in terms of our understanding of its multifunctional modes of action, research is moving closer to the design of approaches directed toward modulating its activities for therapeutic benefit. PMID- 16257257 TI - Industrial relevance of thermophilic Archaea. AB - The dramatic increase of newly isolated extremophilic microorganisms, analysis of their genomes and investigations of their enzymes by academic and industrial laboratories demonstrate the great potential of extremophiles in industrial (white) biotechnology. Enzymes derived from extremophiles (extremozymes) are superior to the traditional catalysts because they can perform industrial processes even under harsh conditions, under which conventional proteins are completely denatured. In particular, enzymes from thermophilic and hyperthermophilic Archaea have industrial relevance. Despite intensive investigations, our knowledge of the structure-function relationships of their enzymes is still limited. Information concerning the molecular properties of their enzymes and genes has to be obtained to be able to understand the mechanisms that are responsible for catalytic activity and stability at the boiling point of water. PMID- 16257258 TI - Protein transport in Archaea: Sec and twin arginine translocation pathways. AB - The transport of proteins into and across hydrophobic membranes is an essential cellular process. The majority of proteins that are translocated in an unfolded conformation traverse the membrane by way of the universally conserved Sec pathway, whereas the twin arginine translocation pathway is responsible for the transport of folded proteins across the membrane. Structural, biochemical and genetic analyses of these processes in Archaea have revealed unique archaeal features, and have also provided a better understanding of these pathways in organisms of all domains. Further study of these pathways in Archaea might elucidate fundamental principles involved in each type of transport and could help determine their relative costs and benefits as well as evolutionary adaptations in protein secretion strategies. PMID- 16257259 TI - Elevation of serum thioredoxin levels in patients with nonalcoholic steatohepatitis. AB - Thioredoxin (TRX) is induced by many oxidative stresses. Serum TRX levels were significantly elevated in nonalcoholic steatohepatitis (NASH) patients, as compared to simple fatty liver (FL) patients or healthy controls. Serum TRX levels in NASH patients were significantly correlated with serum ferritin levels, but not with other variables. Removal of hepatic excess iron by phlebotomy significantly decreased the serum levels of TRX and ALT in NASH patient. Therefore, the pathogenesis of NASH may be associated with iron-related oxidative stress. The serum TRX level is a parameter for discriminating NASH from FL. PMID- 16257261 TI - ARCHITECT anti-HCV assay. PMID- 16257260 TI - Evidence of human coronavirus HKU1 and human bocavirus in Australian children. AB - Undiagnosed cases of respiratory tract disease suspected of an infectious aetiology peak during the winter months. Since studies applying molecular diagnostic assays usually report reductions in the number of undiagnosed cases of infectious disease compared to traditional techniques, we applied PCR assays to investigate the role of two recently described viruses, namely human coronavirus (HCoV) HKU1 and human bocavirus (HBoV), in a hospital-based paediatric population. Both viruses were found among Australia children with upper or lower respiratory tract disease during the autumn and winter of 2004, contributing to 21.1% of all microbial diagnoses, with individual incidences of 3.1% (HCoV-HKU1) and 5.6% (HBoV) among 324 specimens. HBoV was found to coincide with another virus in more than half of all instances and displayed a single genetic lineage, whilst HCoV-HKU1 was more likely to occur in the absence of another microbe and strains could be divided into two genetic lineages which we propose be termed HCoV-HKU1 type A and type B. Children under the age of 2 years were most at risk of infection by these viruses which contribute significantly to the microbial burden among patients with respiratory tract disease during the colder months. PMID- 16257262 TI - Fatty acid utilisation and metabolism in caecal enterocytes of rainbow trout (Oncorhynchus mykiss) fed dietary fish or copepod oil. AB - A combined fatty acid metabolism assay was employed to determine fatty acid uptake and relative utilisation in enterocytes isolated from the pyloric caeca of rainbow trout. In addition, the effect of a diet high in long-chain monoenoic fatty alcohols present as wax esters in oil derived from Calanus finmarchicus, compared to a standard fish oil diet, on caecal enterocyte fatty acid metabolism was investigated. The diets were fed for 8 weeks before caecal enterocytes from each dietary group were isolated and incubated with [1-14C]fatty acids: 16:0, 18:1n-9, 18:2n-6, 18:3n-3, 20:1n-9, 20:4n-6, 20:5n-3, and 22:6n-3. Uptake was measured over 2 h with relative utilisation of different [1-14C]fatty acids calculated as a percentage of uptake. Differences in uptake were observed, with 18:1n-9 and 18:2n-6 showing the highest rates. Esterification into cellular lipids was highest with 16:0 and C18 fatty acids, accounting for over one-third of total uptake, through predominant incorporation in triacylglycerol (TAG). The overall utilisation of fatty acids in phospholipid synthesis was low, but highest with 16:0, the most prevalent fatty acid recovered in intracellular phosphatidylcholine (PC) and phosphatidylinositol (PI), although exported PC exhibited higher proportions of C20/C22 polyunsaturated fatty acids (PUFA). Other than 16:0, incorporation into PC and PI was highest with C20/C22 PUFA and 20:4n-6 respectively. Recovery of labelled 18:1n-9 in exported TAG was 3-fold greater than any other fatty acid which could be due to multiple esterification on the glycerol 'backbone' and/or increased export. Approximately 20-40% of fatty acids taken up were beta-oxidised, and was highest with 20:4n-6. Oxidation of 20:5n-3 and 22:6n-3 was also surprisingly high, although 22:6n-3 oxidation was mainly attributed to retroconversion to 20:5n-3. Metabolic modification of fatty acids by elongation-desaturation was generally low at <10% of [1-14C]fatty acid uptake. Dietary copepod oil had generally little effect on fatty acid metabolism in enterocytes, although it stimulated the elongation and desaturation of 16:0 and elongation of 18:1n-9, with radioactivity recovered in longer n-9 monoenes. The monoenoic fatty acid, 20:1n-9, abundant in copepod oil as the homologous alcohol, was poorly utilised with 80% of uptake remaining unesterified in the enterocyte. However, the fatty acid composition of pyloric caeca was not influenced by dietary copepod oil. PMID- 16257264 TI - Selection and characteristics of peptides that bind thermally grown silicon dioxide films. AB - Dodecapeptides with affinity for thermally grown silicon dioxide were isolated by phage display. Selectants had high histidine content, though distributions of histidine are distinct from reported silica particle-precipitating peptides. Our peptides will have utility as nanoscale affinity domains when inserted into proteins intended for deposition on thermal oxide surfaces/interfaces in micro/nanodevices. PMID- 16257263 TI - Two highly homologous phospholipase D isoenzymes from Papaver somniferum L. with different transphosphatidylation potential. AB - The genes of two phospholipase D (PLD) isoenzymes, PLD1 and PLD2, from poppy seedlings (2829 and 2828 bp) were completely sequenced. The two genes have 96.9% identity in the encoding region and can be assigned to the alpha-type of plant PLDs. The corresponding amino acid sequences do not contain any signal sequences. One Asn-glycosylation site, six and two phosphorylation sites for protein kinase C and tyrosine kinase, respectively, and two phosphatidylinositol-4,5 bisphosphate binding motifs could be identified. Like in most plant PLDs, two HKD motifs and one C2 domain are present. PLD1 and PLD2 have ten and nine cysteine residues. The two enzymes were expressed in E. coli and purified to homogeneity by Ca2+ ion-mediated hydrophobic interaction chromatography. The Ca2+ ion concentration needed for carrier binding of the two enzymes in chromatography as well as for optimum activity was found to be considerably higher (>100 mM) than with other alpha-type plant PLDs. Although PLD1 and PLD2 differ in eleven amino acids only, they showed remarkable differences in their transphosphatidylation activity. Two amino acid exchanges within and near the first HKD motif contribute to this difference as shown by the A349E/E352Q-variant of PLD2. PMID- 16257265 TI - Innate immunity to mycobacterial infection in mice: critical role for toll-like receptors. AB - Toll-like receptors (TLRs) play a critical role in the recognition of several pathogens, including Mycobacterium tuberculosis. Mycobacterial antigens recognize distinct TLRs resulting in rapid activation of cells of the innate immune system. Ablation of most of the TLR signalling as in mice deficient for the common adaptor protein MyD88 reveals that TLR is crucial for the activation of an innate immune response. MyD88-deficient mice are unable to clear virulent mycobacteria and succumb to acute necrotic pneumonia. Despite the profound defect of the innate immune response, MyD88 deficiency allows the emergence of an adaptive immunity. These data demonstrate that activation of multiple TLRs contributes to an efficient innate response to mycobacteria, while MyD88-dependent signalling is dispensable to generate adaptive immunity. PMID- 16257266 TI - Progressive loss of renal function is associated with activation and depletion of naive T lymphocytes. AB - We hypothesized that progressive loss of renal function specifically affects certain T cell subsets. T lymphocyte subsets of patients with chronic kidney disease and healthy controls were characterized by flow cytometry using heparin anticoagulated whole blood samples. Plasma interleukin (IL)-7 and IL-15 concentrations were determined as these cytokines are critically involved in T cell homeostasis. The results revealed that a progressive decrease in renal function is associated with activation and selective loss of naive T cells and CD4+ central memory cells and a marked increase in CD8+ memory T cells that lack CD45RO and CCR7. The profile of T cell subsets of patients with CKD 5 with or without hemodialysis treatment was similar except for a pronounced shift to Th1 cells in hemodialysis patients. IL-7 but not IL-15 plasma concentrations were lowered in patients with end-stage renal disease as compared to healthy controls. PMID- 16257267 TI - HLA B44 is associated with decreased severity of autoimmune lymphoproliferative syndrome in patients with CD95 defects (ALPS type Ia). AB - Autoimmune lymphoproliferative syndrome (ALPS) is a disorder of lymphocyte apoptosis characterized by non-malignant lymphadenopathy and splenomegaly, expansion of T cells without either CD4 or CD8 surface markers, and increased incidence of autoimmune diseases and lymphoma. Most patients with ALPS have dominant, heterozygous mutations in tumor necrosis factor receptor superfamily member 6 (TNFRSF6), which encodes CD95, also known as Fas, a mediator of apoptosis. Penetrance and range of disease manifestations in ALPS are highly variable, even among family members who share the same dominant TNFRSF6 mutation. To evaluate HLA as a candidate modifier locus, we typed HLA A, B (including subtypes), and DQB alleles in 356 individuals from 63 unrelated families with defined TNFRSF6 mutations associated with ALPS. We also developed a quantitative severity score and performed statistical analysis. Among the healthier, mutation bearing individuals, transmission of HLA B44 was significantly overrepresented (nominal P<0.0074) as compared to transmission in patients with severe clinical features of ALPS. The B44 allele may exert a protective role in ALPS. PMID- 16257268 TI - Amelioration of SLE-like manifestations in (NZBxNZW)F1 mice following treatment with a peptide based on the complementarity determining region 1 of an autoantibody is associated with a down-regulation of apoptosis and of the pro apoptotic factor JNK kinase. AB - Treatment with peptides based on the complementarity determining regions (CDR) of murine and human monoclonal anti-DNA antibodies that bear the common idiotype, 16/6 Id, ameliorates disease manifestations of mice with either induced or spontaneous SLE. Aberrant expression and function of the p21Ras/MAP kinase pathway are associated with active SLE. Therefore, we examined the effect of treatment with a CDR1-based peptide of a human autoantibody (hCDR1) on the p21Ras pathway and SLE manifestations of SLE-prone (NZBxNZW)F1 mice. Untreated SLE afflicted mice demonstrated increased expression of p21Ras and the phosphorylated active form of its down-stream element JNK kinase in conjunction with reduced hSOS and unchanged p120GAP, as compared to healthy controls. Amelioration of SLE manifestations following treatment with hCDR1 was associated with a diminished expression of phosphorylated JNK kinase, mainly in the T cell population that also exhibited reduced rates of apoptosis. Thus, hCDR1 therapy ameliorates SLE, at least in part, via down-regulation of the activity of the pro-apoptotic JNK kinase. PMID- 16257269 TI - Hyperglycemia and reactive oxygen species mediate apoptosis in aortic endothelial cells through Janus kinase 2. AB - The generation of reactive oxygen species (ROS) has been implicated in the perturbation of endothelial function and cell death. However, the specific signaling pathways which mediate and modifying this response have not been fully elucidated. Therefore, in this study we tested the hypothesis that activation of JAK2 is involved in the aortic endothelial cell (EC) response to ROS. When ECs were exposed to HG (25 mM) for 6 h or ROS (i.e., H(2)O(2) (100 microM)) for 1 h and returned to normal medium we found a decrease in cell density and morphologic signs of apoptosis. Furthermore, incubation of ECs with HG and H(2)O(2) also resulted in the tyrosine phosphorylation of JAK2. In addition, pretreatment of ECs with AG-490, an inhibitor of JAK2, prevented nuclear fragmentation, whereas inhibitors of Jun kinase (SP 600125), MAP kinase (PD 98059), Src kinase (PP2) or PI-3 kinase (wortmannin) were without effect. Finally, immunoblot analysis of caspase-3 and PARP cleavage confirmed a role for activation of JAK2 in both HG- or ROS-induced apoptosis, based on inhibition by either AG-490 or adenoviral transfection with a dominant-negative JAK2 mutant. In conclusion the activation of JAK2 plays a pivotal role in oxidant stress-induced commitment of ECs to apoptosis, based on studies with HG and H(2)O(2). PMID- 16257271 TI - Evaluation of nasal barrier dysfunction at acute- and late-phase reactions in a guinea pig model of allergic rhinitis. AB - Allergic rhinitis is a common disease characterized by the symptoms of pruritus, sneezing, hypersecretion and nasal blockage. Increased mucosal barrier permeability has been suggested to be an indicator for the severity of allergic rhinitis. This study investigates the passage of radiolabelled albumin from the nasal mucosal circulation into the lumen in guinea pigs intraperitoneally sensitized and intranasally challenged with antigen. In order to characterize the allergic rhinitis model, we evaluated a number of potential influencing factors in nasal plasma exudation, including antigen doses, volumes of antigen solution used, and animal position during the nasal lavage, and the conditions of nasal lavage. The number of eosinophils and levels of histamine and leukotriene B4 in the nasal lavage and eosinophils in the nasal mucosa were determined at the early and late phases after antigen challenge. We also compared the effects of topical nasal treatments for allergic rhinitis on nasal inflammatory responses. Our results demonstrate that, in the guinea pig nasal mucosa, topical challenge with antigens induces plasma exudation and histamine release at the acute-phase reaction, and plasma exudation and eosinophil infiltration at the late-phase reaction. These changes are similar to those reported in human allergic rhinitis. Alterations of nasal plasma exudation, histamine release and eosinophil influx were dependent upon the concentrations and volumes of antigens. An antihistamine inhibited the acute-phase reaction partially, whereas budesonide inhibited effects at the late-phase reaction. We suggest that this model of guinea pig allergic rhinitis with the early and late responses may be useful for high throughout screening of new drugs. PMID- 16257270 TI - Jak2 tyrosine kinase prevents angiotensin II-mediated inositol 1,4,5 trisphosphate receptor degradation. AB - In addition to its role as a vasoconstrictor, angiotensin II also acts as a potent growth factor by activating several tyrosine kinases, including Jak2. Interestingly, Jak2 has been linked to similar cardiovascular pathologies as have been previously linked to the renin-angiotensin system. Identifying the downstream targets of Jak2 via the AT(1) receptor may therefore elucidate its role in the progression of various pathologies. Previously, microarray analysis from our laboratory identified the Type 1 inositol 1,4,5 trisphosphate (IP(3)) receptor as a potential target of Jak2 following chronic stimulation by angiotensin II. Therefore, we hypothesized that Jak2 regulates IP(3) receptor expression in response to angiotensin II. To test this hypothesis, rat aortic smooth muscle (RASM) cells over-expressing a dominant negative (DN) Jak2 protein were used. The Jak2-dependent signaling in these cells is reduced approximately 90% when compared to RASM control cells. Analysis of protein expression showed that the IP(3) receptor was degraded approximately 2-fold (P<0.05) in cells lacking functional Jak2 within 1 h of treatment by angiotensin II. Notably, degradation of the IP(3) receptor was reversible since protein levels were restored to normal following 2 h of recovery from angiotensin II. To eliminate the possibility of clonal artifact in the DN cells, wild type RASM cells were treated with the Jak2 pharmacological inhibitor, AG490. We found that angiotensin II treatment degraded IP(3) receptor in AG490-treated cells, but not in the vehicle controls. Treatment with lactacystin, a proteasome inhibitor, completely blocked angiotensin II-mediated degradation of IP(3) receptor, thereby suggesting that the degradation occurs through a proteasome-dependent mechanism. Moreover, the degradation of IP(3) receptor in DN cells correlated with a significant loss of intracellular calcium mobilization when treated with angiotensin II (DN 27.4+/ 1.1% vs. WT 42.2+/-4.7%; n=5, P=0.002). We next examined through what mechanism Jak2 regulates the IP(3) receptor. When wild type RASM cells were treated with PP2, an Src-family inhibitor, IP(3) receptor expression was markedly reduced. Since previous data show that Fyn, a downstream target of Jak2, is able to phosphorylate the IP(3) receptor at Tyr 353, we believe our data suggest that Jak2 prevents the angiotensin II-mediated IP(3) receptor degradation through the activation of Fyn. In conclusion, these data suggest that Jak2 has a protective role in maintaining IP(3) receptor expression, potentially through activation of Fyn and subsequent phosphorylation of the IP(3) receptor. PMID- 16257272 TI - Harnessing the power of gene microarrays for the study of brain aging and Alzheimer's disease: statistical reliability and functional correlation. AB - During normal brain aging, numerous alterations develop in the physiology, biochemistry and structure of neurons and glia. Aging changes occur in most brain regions and, in the hippocampus, have been linked to declining cognitive performance in both humans and animals. Age-related changes in hippocampal regions also may be harbingers of more severe decrements to come from neurodegenerative disorders such as Alzheimer's disease (AD). However, unraveling the mechanisms underlying brain aging, AD and impaired function has been difficult because of the complexity of the networks that drive these aging related changes. Gene microarray technology allows massively parallel analysis of most genes expressed in a tissue, and therefore is an important new research tool that potentially can provide the investigative power needed to address the complexity of brain aging/neurodegenerative processes. However, along with this new analytic power, microarrays bring several major bioinformatics and resource problems that frequently hinder the optimal application of this technology. In particular, microarray analyses generate extremely large and unwieldy data sets and are subject to high false positive and false negative rates. Concerns also have been raised regarding their accuracy and uniformity. Furthermore, microarray analyses can result in long lists of altered genes, most of which may be difficult to evaluate for functional relevance. These and other problems have led to some skepticism regarding the reliability and functional usefulness of microarray data and to a general view that microarray data should be validated by an independent method. Given recent progress, however, we suggest that the major problem for current microarray research is no longer validity of expression measurements, but rather, the reliability of inferences from the data, an issue more appropriately redressed by statistical approaches than by validation with a separate method. If tested using statistically defined criteria for reliability/significance, microarray data do not appear a priori to require more independent validation than data obtained by any other method. In fact, because of added confidence from co-regulation, they may require less. In this article we also discuss our strategy of statistically correlating individual gene expression with biologically important endpoints designed to address the problem of evaluating functional relevance. We also review how work by ourselves and others with this powerful technology is leading to new insights into the complex processes of brain aging and AD, and to novel, more comprehensive models of aging related brain change. PMID- 16257273 TI - Evidence for Escherichia coli polymerase II mutagenic bypass of intrastrand DNA crosslinks. AB - The mutagenic potentials of DNAs containing site- and stereospecific intrastrand DNA crosslinks were evaluated in Escherichia coli cells that contained a full complement of DNA polymerases or were deficient in either polymerases II, IV, or V. Crosslinks were made between adjacent N(6)-N(6) adenines and consisted of R,R- and S,S-butadiene crosslinks and unfunctionalized 2-, 3-, and 4-carbon tethers. Although replication of single-stranded DNAs containing the unfunctionalized 3- and 4-carbon tethers were non-mutagenic in all strains tested, replication past all the other intrastrand crosslinks was mutagenic in all E. coli strains, except the one deficient in polymerase II in which no mutations were ever detected. However, when mutagenesis was analyzed in cells induced for SOS, mutations were not detected, suggesting a possible change in the overall fidelity of polymerase II under SOS conditions. These data suggest that DNA polymerase II is responsible for the in vivo mutagenic bypass of these lesions in wild-type E. coli. PMID- 16257274 TI - Introducing the need for lung transplantation in children with cystic fibrosis: parental experiences. AB - Lung transplantation (LTx) is the only treatment available for adult and pediatric end-stage lung disease secondary to cystic fibrosis (CF). The timing of introducing LTx has significant medical and psychological implications for the child and the family. This study explored the views and recommendations of parents of children with CF, who had been asked to consider LTx and referred to a national transplant centre. Parents participated in a telephone-based, semi structured interview. Responses were analysed using Content Analysis. Parental recommendations and the emergent protocol are discussed, together with implications for clinical practice. PMID- 16257275 TI - Analysis of microperoxidases using liquid chromatography, post-column substrate conversion and fluorescence detection. AB - A liquid chromatographic method with on-line activity determination for microperoxidases has been developed. After enzymatic digestion of a cytochrome, possibly under formation of microperoxidases, the product mixture is separated by reversed-phase liquid chromatography. The products first pass a diode-array detector, and are then subjected to a reaction with 4-(N-methylhydrazino)-7-nitro 2,1,3-benzooxadiazole (MNBDH) and hydrogen peroxide. In a reaction coil, microperoxidases catalyze the reaction under formation of the fluorescent 4-(N methylamino)-7-nitro-2,1,3-benzooxadiazole (MNBDA). Quantification of the microperoxidases is performed using a fluorescence detector at an excitation wavelength of 470 nm and an emission wavelength of 545 nm, respectively. For this LC-based detection system, limits of detection are 3 x 10(-8) mol/L, limits of quantification are 9 x 10(-8) mol/L, and a linear range from 9 x 10(-8) mol/L to 3 x 10(-6) mol/L is obtained for the microperoxidases MP-9 and MP-11. A highly active microperoxidase MP-6 was found in the reaction of cytochrome c from bovine heart with protease from streptomyces griseus. PMID- 16257276 TI - Chlamydia pneumoniae AP endonuclease IV could cleave AP sites of double- and single-stranded DNA. AB - Endonuclease IV gene, the only putative AP endonuclease of C. pneumoniae genome, was cloned into pET28a. Recombinant C. pneumoniae endonuclease I V (CpEndoIV) was expressed in E. coli and purified to homogeneity. CpEndoIV has endonuclease activity against apurinic/apyrimidinic sites (AP sites) of double-stranded (ds) oligonucleotides. AP endonuclease activity of CpEndoIV was promoted by divalent metal ions Mg2+ and Zn2+, and inhibited by EDTA. The natural (A, T, C and G) and modified (U, I and 8-oxo-G (GO)) bases opposite AP site had little effect on the cleavage efficiency of AP site of ds oligonucleotides by CpEndoIV. However, the CpEndoIV-dependent cleavage of AP site opposite modified base GO was strongly inhibited by Chlamydia DNA glycosylase MutY. Interestingly, the AP site in single stranded (ss) oligonucleotides was also the effective substrate of CpEndoIV. Similar to E. coli endonuclease IV, AP endonuclease activity of CpEndoIV was also heat-stable to some extent, with a half time of 5 min at 60 degrees C. PMID- 16257277 TI - -511 C/T IL1B gene polymorphism is associated to resistance to bisphosphonates treatment in Paget disease of bone. AB - INTRODUCTION: Osteoclasts are the most important cells involved in the pathogenesis of Paget disease of bone (PDB). Cytokines stimulate osteoclast differentiation and activation, with some of them over-expressed in pagetic osteoclasts. We have assessed whether genetic variability in genes coding of proteins from the IL1 pathway clustered in chromosome 2 is associated with clinical characteristics and the therapeutic response of patients with PDB. METHODS: We have studied -511 C/T and +3953 T/C polymorphisms of the IL1B gene, a HinfI polymorphism in the 5'UTR of the IL1R1 gene, and a variable number of tandem repeats (VNTR) in the intron 2 of the IL1RN gene, in 165 patients diagnosed as suffering from PDB and in 122 healthy controls. Distribution of genotypes and alleles was studied for association with clinical and laboratory data and response to bisphosphonate (BSP) treatment. RESULTS: No differences were observed in the distribution of genotypes or alleles between PDB patients and control subjects. We also failed to detect differences concerning epidemiological, clinical and laboratory data in the series of PDB patients. However, the -511 CC genotype of the IL1B gene was associated with a higher percentage of resistance to BSP (49% vs. 20%; P = 0.00 for all BSP, 60% vs. 39%, P = 0.17 for etidronate, 50% vs. 37% P = 0.53 for clodronate, 48 vs. 34% P = 0.05 for tiludronate and 50% vs. 4% P = 0.01 for risedronate). CONCLUSIONS: Our results suggest that the -511 CC genotype of the IL1B gene could be related to resistance to bisphosphonates in patients with PDB. PMID- 16257278 TI - Modulation of insulin-like growth factor 1 levels in human osteoarthritic subchondral bone osteoblasts. AB - Human osteoarthritis (OA) is characterized by cartilage loss, bone sclerosis, osteophyte formation and inflammation of the synovial membrane. We previously reported that OA osteoblasts (Ob) show abnormal phenotypic characteristics possibly responsible for bone sclerosis and that two subgroups of OA patients can be identified by low or high endogenous production of prostaglandin E2 (PGE2) by OA Ob. Here, we determined that the elevated PGE2 levels in the high OA subgroup were linked with enhanced cyclooxygenase-2 (COX-2) protein levels compared to normal and low OA Ob. A linear relationship was observed between endogenous PGE2 levels and insulin-like growth factor 1 (IGF-1) levels in OA Ob. As parathyroid hormone (PTH) and PGE2 are known stimulators of IGF-1 production in Ob, we next evaluated their effect in OA Ob. Both subgroups increased their IGF-1 production similarly in response to PGE2, while the high OA subgroup showed a blunted response to PTH compared to the low OA group. Conversely, only the high OA group showed a significant inhibition of IGF-1 production when PGE2 synthesis was reduced with Naproxen, a non-steroidal antiinflammatory drug (NSAID) that inhibits cyclooxygenases (COX). The PGE2-dependent stimulation of IGF-1 synthesis was due in part to the cAMP/protein kinase A pathway since both the direct inhibition of this pathway with H-89 and the inhibition of EP2 or EP4 receptors, linked to cAMP production, reduced IGF-1 synthesis. The production of the most abundant IGF-1 binding proteins (IGFBPs) in bone tissue, IGFBP-3, -4, and -5, was lower in OA compared to normal Ob independently of the OA group. Under basal condition, OA Ob expressed similar IGF-1 mRNA to normal Ob; however, PGE2 stimulated IGF-1 mRNA expression more in OA than normal Ob. These data suggest that increased IGF-1 levels correlate with elevated endogenous PGE2 levels in OA Ob and that higher IGF-1 levels in OA Ob could be important for bone sclerosis in OA. PMID- 16257279 TI - Bone mineral density among female sports participants. AB - Training for and participation in impact-loading sports are associated with alterations in bone strength which are specific to anatomical site and type of strain. The effect of exercise on bone mineral density (BMD) depends on the type of activity engaged in. Sports with high impact loading seem to have a positive effect in promoting bone mineralisation, whereas those with low impacts may have negative or no effects. The aims of the present study were to compare BMD and body composition measures among female participants in three distinctly different sports and investigate differences from sedentary control subjects. Participants were club and university level Rugby Union football players (n = 30, age: 21.4 +/ 1.9 years, height: 1.67 +/- 0.05 m, mass: 73.3 +/- 10.7 kg), netball players (n = 20, 20.7 +/- 1.3 years, 1.68 +/- 0.07 m, 64.3 +/- 7.2 kg), distance runners (n = 11, 21.5 +/- 2.6 years, 1.68 +/- 0.04 m, 57.1 +/- 6.1 kg), and sedentary controls (n = 25, 21.4 +/- 1.1 years; 1.64 +/- 0.07 m, 56.8 +/- 6.8 kg). With the exception of three distance runners, all participants were eumenorrhoeic. Bone mineral density scans were performed for whole-body, left proximal femur, and lumbar spine (L1-4) using dual-energy X-ray absorptiometry. Fat mass, percent body fat, and fat-free soft tissue mass were assessed from whole-body scans. Regional and segmental analysis was also carried out on whole-body BMD data using standard procedures. The runners had a lower fat mass and percent body fat compared to the other sports participants and the controls. All sports groups had higher BMD values than had the controls. Density of bone in the upper body was most pronounced in the rugby football players and least pronounced in the runners. Positive effects were evident at all sites for the rugby players. There were significant correlations between BMD and fat-free soft tissue mass, BMD and body mass, and BMD and training volume. It is concluded that sports participation has positive effects on BMD. The effects are site-specific and depend on the loading characteristics of the sport. PMID- 16257280 TI - Angiotensin converting enzyme inhibitor use is associated with higher bone mineral density in elderly Chinese. AB - Hypertension and osteoporosis are two major chronic diseases affecting the elderly. A cross-sectional study of 3887 Chinese men (n = 1958) and women (n = 1929) was used to explore the association between angiotensin converting enzyme inhibitor (ACEI) use and bone mineral density (BMD). The participants were aged 65 years and above, and were recruited using a combination of private solicitation and public advertising from community centers, housing estates, and the general community in Hong Kong. Demographic, medical, and lifestyle information was obtained from face to face interviews using standardized questionnaire, and physical examination measurements included anthropometry, tibial, and brachial systolic blood pressures, femoral neck, total hip, and lumbar spine BMD. In multiple regression analyses, after adjusting for age, weight, height, thiazide, beta-blocker, calcium channel blocker, statin, corticosteroid, and calcium supplement use, history of diabetes, heart disease, peripheral vascular disease, cigarette smoking, alcohol intake, and physical activity level, ACEI use was associated with higher femoral neck BMD (+0.015 g/cm2, P = 0.035) in women, and higher femoral neck (+0.015 g/cm2, P = 0.017), total hip (+0.016 g/cm2, P = 0.021), and lumbar spine (+0.043 g/cm2, P < 0.001) BMD in men. Thiazide use was associated with higher BMD at all three sites in general, although associations with BMD increase at the total hip (P = 0.07) and femoral neck (P = 0.09) were weak in men. Calcium channel blocker use was only significantly associated with BMD increase at the lumbar spine (P = 0.03) in women, and beta-blocker use did not have significant associations with BMD at any site. This study suggests that in addition to thiazide diuretics ACEI may have possible benefits in treating not only hypertension but also osteoporosis among older Chinese. PMID- 16257281 TI - Liver-derived IGF-I is permissive for ovariectomy-induced trabecular bone loss. AB - INTRODUCTION: Estrogen deficiency results in trabecular bone loss, associated with T-cell proliferation in the bone marrow. Insulin-like growth factor I (IGF I) is involved in the regulation of both bone metabolism and lymphopoiesis. A major part of serum IGF-I is derived from the liver. The aim of the present study was to investigate the role of liver-derived IGF-I for ovariectomy (ovx)-induced trabecular bone loss. MATERIALS AND METHODS: Mice with adult liver-specific IGF-I inactivation (LI-IGF-I-/-) and wild type mice (WT) were either ovx or sham operated. After 5 weeks, the skeletal phenotype was analyzed by pQCT and microCT. The bone marrow cellularity was analyzed using FACS technique, and mRNA levels were quantified using real-time PCR. RESULTS: Ovx resulted in a pronounced reduction in trabecular bone mineral density (-52%, P < 0.001), number (-45%, P < 0.01) and thickness (-13%, P < 0.01) in WT mice while these bone parameters were unaffected by ovx in LI-IGF-I-/- mice. Furthermore, ovx increased the number of T cells in the bone marrow of the femur in WT but not in LI-IGF-I-/- mice. Interleukin 7 (IL-7) has been reported to stimulate the formation and function of osteoclasts by inducing the expression of receptor activator of NF-kappaB ligand (RANKL) on T-cells. IL-7 mRNA levels and the RANKL/osteoprotegerin ratio in bone were increased by ovx in WT but not in LI-IGF-I-/- mice. CONCLUSIONS: Liver derived IGF-I is permissive for ovx-induced trabecular bone loss. Our studies indicate that IGF-I might exert this permissive action by modulation of the number of T-cells and the expression of IL-7, which in turn is of importance for the RANKL/OPG ratio and consequently osteoclastogenesis in the bone marrow. PMID- 16257282 TI - Surfactant enhanced liquid-phase microextraction of basic drugs of abuse in hair combined with high performance liquid chromatography. AB - The aim of this study was to evaluate the performance of a technique for simultaneous testing of hydrophilic abuse drugs in hair. The analysis of, codeine and methadone in morphine hair included incubation in methanol (5h, 50 degrees C), Surfactant enhanced liquid-phase microextraction (SE-LPME) and HPLC analysis. This study has demonstrated that SE-LPME constitute a real alternative to the other liquid-phase microextraction methods, for pre-concentration and extraction of hydrophilic drugs in biological samples and has shown the advantages of these optimized methodologies over the traditional microextraction techniques. For these drugs recoveries in the range of 57.5-93.7 were obtained from hair. The drugs were enriched by a factor of 61-128 during SE-LPME. Linearity (r2, 0.9982 0.9997) was obtained in the range of 50-500 microg/l for morphine and 10-500 microg/l for codeine and methadone. PMID- 16257283 TI - Dual-phase twisters: a new approach to headspace sorptive extraction and stir bar sorptive extraction. AB - The fields of applicability of headspace sorptive extraction (HSSE) and stir bar sorptive extraction (SBSE) using polydimethylsiloxane (PDMS) as sorbent have been intensively discussed and widely described. One of the limits of sorptive extraction is that PDMS (i.e. an apolar phase) is the only polymer currently in use making it difficult to recover polar analytes from complex or multi ingredient matrices and those with very volatile components (C1-C4 analytes). Dual-phase twisters are here introduced as new tools for HSSE and SBSE to overcome the above limits. Dual-phase twisters combine the concentration capabilities of two or more sampling materials operating in different ways (in this case sorption and adsorption). The new twisters consist of a short PDMS tube the ends of which are closed with two magnetic stoppers, thus creating an inner cavity that can be packed with different types of adsorbents like activated carbons. The concentration capability of dual-phase twisters was evaluated by using them for the HSSE and SBSE sampling of a number of matrices in the vegetable, food and environmental fields. The contributions made by different carbons to recovery, repeatability and intermediate precision were also investigated. PMID- 16257284 TI - Stability of benzalkonium surfactants on hemimicelle-based solid-phase extraction cartridges. AB - The capability of hemimicelle-based solid-phase extraction cartridges for the preservation of organic compounds after their concentration from water samples was investigated for the first time. The approach is illustrated by studying the stability of benzalkonium homologue (C12, C14 and C16) surfactants (BAS) on monolayers of dodecyl sulphate (SDS) hemimicelles formed on alumina. The stability study included storage of cartridges at room temperature, at 4 and -20 degrees C, during a period of up to 3 months. The influence of water matrix components was also investigated from parallel experiments using spiked distilled, river and wastewater samples. Complete recovery of BAS was obtained for all storage conditions tested. Recoveries were independent on the alkyl chain length of BAS homologues and water matrix. The SPE of BAS on the SDS hemimicelles had a strong stabilizing effect for the target compounds and their analysis can be accomplished after at least 3 months without the necessity of special storage conditions for cartridges. Because of the lack of data, an additional stability study was carried out for BAS in an aqueous matrix using traditional preservation methods such as acidification (pH 2)/refrigeration, addition of formaldehyde (5%)/refrigeration, and freezing (-20 degrees C). Only combination of chemical addition (e.g. nitric acid or formaldehyde)/refrigeration was found effective to preserve BAS in the short term (e.g. for a week), then losses up to 40% were observed for these target compounds after a month. PMID- 16257285 TI - Emerging approaches to estimate retention factors in high performance liquid chromatography. AB - The retention factor is one of the most universally used parameters in chromatography. The errors associated with the conventional ways to determine the retention factor of compounds in liquid chromatography are studied and compared with those corresponding to new approaches. The later avoid the use of extra column time and hold-up time values, which have proven to be tedious and ambiguous. Simulations and real data, used to examine the accuracy of four different approaches (two classic and two new), suggest that the new approaches could be considered more satisfactory than the classic ones. PMID- 16257286 TI - Varietal differences among the anthocyanin profiles of 50 red table grape cultivars studied by high performance liquid chromatography. AB - In order to develop a method that allows to distinguish between grape cultivars, the anthocyanin profiles of 50 accessions from the "Mision Biologica de Galicia" germplasm collection were studied by high performance liquid chromatography (HPLC). Nineteen anthocyanins were totally or partly identified and significant quantitative differences between the studied anthocyanin markers were found. With this method all 50 cultivars examinated could be easily distinguished from each other. In addition, the HPLC fingerprints and the relative-area anthocyanins plot for every cultivar has been elaborated and stored in a database. To test the validity of this method, several unknown samples have been analysed comparing their anthocyanin profile with the fingerprint database, and we may conclude that this has been proved to be of great value for grape cultivar recognition. PMID- 16257287 TI - Liquid chromatography of polymers under limiting conditions of desorption II. Tandem injection and quantitative molar mass determination. AB - Liquid chromatography under limiting conditions of desorption (LC LCD) is a method which allows molar mass independent elution of various synthetic polymers. A narrow, slowly moving zone of small molecules, which promotes full adsorption of one kind of polymer species within column (an adsorli) acts as an impermeable barrier for the fast moving macromolecules. The latter accumulate on the barrier edge and elute nearly in total volume of liquid within column. At the same time, transport of less adsorptive macromolecules is not hampered so that these are eluted in the size exclusion (SEC) mode. As result, polymers differing in their polarity and adsorptivity can be easily separated without molar mass interference. Three methods of barrier creation are discussed and compared. It is shown that a fraction of sample may elute unretained if the adsorli sample solvent is used as a barrier in connection with a narrow-pore column packing. One part of excluded macromolecules likely breaks-out from the adsorli zone and this results in partial loss of sample and distortion of the LC LCD peaks. This problem can be avoided if the adsorli zone is injected immediately before sample solution. Applicability of the LC LCD method for polymer separation has been demonstrated with a model mixture of poly(methyl methacrylate) (adsorbing polymer) and polystyrene (non adsorbing polymer) using bare silica gel as a column packing with a combination of tetrahydrofuran (a desorption promoting liquid -a desorli) and toluene (adsorli). It has been shown that the LC LCD procedure with tandem injection allows simple and fast discrimination of polymer blend components with good repeatability and high sample recovery. For quantitative determination of molar masses of both LC LCD and SEC eluted polymers, an additional size exclusion chromatographic column can be applied either in a conventional way or in combination with a multi-angle light scattering detector. A single eluent is used in the latter column, which separates the mixed mobile phase, system peaks and the desorli zone from the polymer peaks so that measurements are free from disturbances caused by the changing eluent composition. The resulting LC LCD x SEC procedure has been successfully applied to poly(methyl methacrylate) samples. PMID- 16257288 TI - Arginine as an effective additive in gel permeation chromatography. AB - A major problem in gel permeation chromatography (GPC) or size exclusion chromatography is non-specific binding of applied proteins to the column matrix (stationary phase). We have tested an aqueous arginine solution as the GPC mobile phase on silica-based and polymer-based columns, using mouse monoclonal antibody and recombinant human activin, interleukin-6, basic fibroblast growth factor, and interferon-gamma as model proteins. We observed that addition of arginine to the mobile phase improves separation of the proteins and their soluble aggregates from the GPC columns, which suggests that arginine is an effective additive for the GPC mobile phase. PMID- 16257289 TI - Determination of novel complexing agents in pulp and paper mill effluents and in lake water by liquid chromatography. AB - A simple and sensitive high performance liquid chromatographic (HPLC) method with UV detection was developed for the determination of new diethanolamine derivative complexing agents. The quantitation of three complexing agents was accomplished as their Cu(II) complexes and their separation on a reversed phase C18 column using 0.01 M tetrabutyl ammoniumhydroxide and methanol as isocratic mobile phase. The limit of detection (LOD) ranges from 0.1 to 2.0 mg/l and the method has a wide linear range, from LOD to 150 mg/l. The method was successfully applied for the analysis of lake water and wastewater effluent from pulp and paper mill spiked with studied complexing agents. PMID- 16257290 TI - Determination of putrescine and cadaverine in seafood (finfish and shellfish) by liquid chromatography using pyrene excimer fluorescence. AB - A liquid chromatography (LC) method is described for the easy determination of the biogenic diamines putrescine (PUT) and cadaverine (CAD) in canned tuna, frozen tuna loin, fresh mahimahi fillet, frozen raw shrimp, cooked lump crabmeat, and fresh and cold-smoked salmon. The method is also a useful screen for histamine (HTA). The method involves homogenization of fish tissue, extraction of biogenic amines into borate-trichloroacetic acid solution, centrifugation, and derivatization of supernatant with 1-pyrenebutanoic acid succinimidyl ester. The derivatized diamine species allow for the intramolecular excimer fluorescence of the pyrene moiety at a higher emission wavelength than is possible for the endogenous tissue monoamines, thus providing visual specificity of detection. All seafood species were fortified with 0.5, 1.0, 5.0, 10.0, and 15.0 microg/g (ppm) of PUT and CAD. Determination was based on standard graphs for PUT and CAD using peak areas with standard solutions equivalent to 0.375, 1.0, 5.0, 10.0, and 20.0 ppm in tissue. A set of five matrix controls (unfortified seafood tissue) were also analyzed; endogenous PUT was found in all samples except the canned tuna, and CAD found only in the shrimp, crab, and cold-smoked salmon. The background amines were thus subtracted prior to determining spike recovery. The intra-assay average recoveries ranged from 71 to 94% across species and spike levels. PMID- 16257291 TI - Solid-phase microextraction coupled with high performance liquid chromatography using on-line diode-array and electrochemical detection for the determination of fenitrothion and its main metabolites in environmental water samples. AB - The aim of this study was to develop a methodology for the analysis of the insecticide fenitrothion and its two main environmental metabolites, fenitrooxon and 3-methyl-4-nitrophenol. For this purpose, a solid-phase microextraction (SPME) method coupled to high performance liquid chromatography (LC) was optimized. Two on-line detectors, diode array (DAD) and direct current amperometrical (DCAD) were used in order to determine sensitivity and selectivity. The effects of the extraction parameters, including exposure and desorption time, pH, temperature, salt concentration and desorption mode on the extraction efficiency were studied. A satisfactory reproducibility for extractions from samples at 20 ppb-level with RSD < 12.5% (n = 10) was obtained. The calibration graphs were linear in the range of 10-1000 microg l(-1) and detection limits for the target compounds were between 1.2 and 11.8 microg l(-1) depending on which detector was used. The method was applied for determining fenitrothion and both its metabolites in river waters which run through forest areas near to aerial application of the pesticide. PMID- 16257292 TI - Simultaneous determination of palladium, platinum, rhodium and gold by on-line solid phase extraction and high performance liquid chromatography with 5-(2 hydroxy-5-nitrophenylazo)thiorhodanine as pre-column derivatization regents. AB - In this paper, 5-(2-hydroxy-5-nitrophenylazo)thiorhodanine (HNATR) was synthesized. A new method for the simultaneous determination of palladium, platinum, rhodium and gold ions as metal-HNATR chelates was developed using a rapid analysis column high performance liquid chromatography equipped with on line solid phase extraction technique. The samples (Water, human urine, geological samples and soil) were digested by microwave acid-digestion. The palladium, platinum, rhodium and gold ions in the digested samples were pre column derivatized with HNATR to form colored chelates. The Pd-HNATR, Pt-HNATR, Rh-HNATR and Au-HNATR chelates can be absorbed onto the front of the enrichment column when they were injected into the injector and sent to the enrichment column [Zorbax Stable Bound, 10 mm x 4.6 mm, 1.8 microm] with a buffer solution of 0.05 mol L(-1) phosphoric acid as mobile phase. After the enrichment had finished, by switching the six ports switching valve, the retained chelates were back-flushed by mobile phase and travelling towards the analytical column. These chelates separation on the analytical column [Zorbax Stable Bound, 10 mm x 4.6 mm, 1.8 microm] was satisfactory with 72% acetonitrile (containing 0.05 mol L(-1) of phosphoric acid and 0.1% of Triton X-100) as mobile phase. The palladium, platinum, rhodium and gold chelates were separated completely within 2.5 min. Compared to the routine chromatographic method, more then 80% of separation time was shortened. By on-line solid phase extraction system, a large volume of sample (10 mL) can be injected, and the sensitivity of the method was greatly improved. The detection limits (S/N=3, the sample injection volume is 10 mL) of palladium, platinum, rhodium and gold in the original samples reaches 1.4, 1.8, 2.0 and 1.2 ng L(-1), respectively. The relative standard deviations for five replicate samples were 2.4-3.6%. The standard recoveries were 88-95%. This method was applied to the determination of palladium, platinum, rhodium and gold in human urine, water and geological samples with good results. PMID- 16257293 TI - Simultaneous quantification of alpha-tocopherol and four major carotenoids in botanical materials by normal phase liquid chromatography-atmospheric pressure chemical ionization-tandem mass spectrometry. AB - A rapid and sensitive method for the simultaneous quantification of alpha tocopherol, beta-carotene, beta-cryptoxanthin, lutein and zeaxanthin in botanical materials has been developed using normal phase liquid chromatography-atmospheric pressure chemical ionization-tandem mass spectrometry (NP-LC-APCI-MS/MS). A systematic extraction procedure for different botanical materials was described and the extraction solvents were matched with normal-phase LC requirement. Quantification was performed by using the external standards and standard calibration curves were linear between 35 and 8505 ng/mL for alpha-tocopherol, 64 15,552 ng/mL for beta-carotene, 40-9720 ng/mL for beta-cryptoxanthin, 70-17,010 ng/mL for lutein and 64-15,552 ng/mL for zeaxanthin with regression coefficient r2 > 0.9996. The limits of quantification (LOQ) were 35, 64, 40, 70 and 66 ng/mL for alpha-tocopherol, beta-carotene, beta-cryptoxanthin, lutein and zeaxanthin, respectively. PMID- 16257294 TI - High-performance liquid chromatography method for the simultaneous determination of thiamine hydrochloride, pyridoxine hydrochloride and cyanocobalamin in pharmaceutical formulations using coulometric electrochemical and ultraviolet detection. AB - The method for the simultaneous determination of thiamine hydrochloride, pyridoxine hydrochloride and cyanocobalamin by high-performance liquid chromatography (HPLC) with coulometric electrochemical and UV detections is presented. The retention time of vitamins was repeatedly determined by isocratic elution using 0.05 M phosphate buffer-10% methanol and 0.018 M trimethylamine (1 ml min(-1), pH 3.55) as mobile phase with the Supelco LC 18 column 5 microm (25 cm x 4.6 mm). The specificity of the method was demonstrated by the retention characteristics, coulometric electrochemical and UV detection. The limits of detection of thiamine, pyridoxine and cyanocobalamin were: 9.2, 2.7 and 0.08 ng/ml, respectively. The method was characterized also by wide concentration range, high sensitivity and good accuracy (99.6-102.7%). The repeatability of the method was evaluated at different level of concentration of vitamins and the relative standard deviation was below 4.5%. The method was successfully applied for the quantification of Vitamins B1, B6 and B12 in pharmaceutical preparations and dietary supplements. PMID- 16257295 TI - High-performance liquid chromatography for quantification of plumbagin, an anti Helicobacter pylori compound of Plumbago zeylanica L. AB - The plant Plumbago zeylanica L. is a semi-climbing shrub that grows throughout Asia and Africa. In our previous study, P. zeylanica L. exhibited high anti Helicobacter pylori and good bactericidal activities over a wide pH range (pH 2 7). Plumbagin - the major ingredient derived from the roots of P. zeylanica L. - is a naphthoquinone compound. In this study, we investigated plumbagin's anti-H. pylori activity and developed a reversed-phase high-performance liquid chromatography (HPLC) method for quantification of plumbagin from P. zeylanica L. We also observed that plumbagin has strong anti-H. pylori activity, with 0.02 0.16 mg/ml as minimum inhibitory concentrations and 0.16-1.28 mg/ml as minimum bactericidal concentrations. Reversed-phase HPLC was performed with a gradient mobile phase composed of water and methanol, and peaks were detected at 254 nm. Standard curves were linearized in the range of from 10 to 200 microg/ml (regression coefficient r2 = 0.99995). After spikes of 50, 100, and 150 microg/ml of plumbagin standard solution, recovery rates were between 97.45 and 99.24%. Both intra- and inter-day precisions had coefficient variation of less than 1% at concentrations of 50, 100, and 150 microg/ml. The limits of detection and quantitation were 0.02 and 0.06 microg/ml, respectively. Based on validation results, this analytical method is a precise, accurate and stable method to quantify plumbagin derived from P. zeylanica L. PMID- 16257296 TI - Characterization of a gel-separated unknown glycoprotein by liquid chromatography/multistage tandem mass spectrometry: analysis of rat brain Thy-1 separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. AB - We developed an efficient and convenient strategy for protein identification and glycosylation analysis of a small amount of unknown glycoprotein in a biological sample. The procedure involves isolation of proteins by electrophoresis and mass spectrometric peptide/glycopeptide mapping by LC/ion trap mass spectrometer. For the complete glycosylation analysis, proteins were extracted in intact form from the gel, and proteinase-digested glycoproteins were then subjected to LC/multistage tandem MS (MSn) incorporating a full mass scan, in-source collision induced dissociation (CID), and data-dependent MSn. The glycopeptides were localized in the peptide/glycopeptide map by using oxonium ions such as HexNAc+ and NeuAc+, generated by in-source CID, and neutral loss by CID-MS/MS. We conducted the search analysis for the glycopeptide identification using search parameters containing a possible glycosylation at the Asn residue with N acetylglucosamine (203 Da). We were able to identify the glycopeptides resulting from predictable digestion with proteinase. The glycopeptides caused by irregular cleavages were not identified by the database search analysis, but their elution positions were localized using oxonium ions produced by in-source CID, and neutral loss by the data-dependent MSn. Then, all glycopeptides could be identified based on the product ion spectra which were sorted from data-dependent CID-MSn spectra acquired around localized positions. Using this strategy, we successfully elucidated site-specific glycosylation of Thy-1, glycosylphosphatidylinositol (GPI)-anchored proteins glycosylated at Asn23, 74, and 98, and at Cys111. High-mannose-type, complex-type, and hybrid-type oligosaccharides were all found to be attached to Asn23, 74 and 98, and four GPI structures could be characterized. Our method is simple, rapid and useful for the characterization of unknown glycoproteins in a complex mixture of proteins. PMID- 16257298 TI - Determination of endocrine-disrupting phenols, acidic pharmaceuticals, and personal-care products in sewage by solid-phase extraction and gas chromatography mass spectrometry. AB - The occurrence, fate, and effects of phenols with endocrine-disrupting properties as well as some pharmaceuticals and personal-care products in the environment have frequently been discussed in recent literature. In many cases, these compounds were determined by using individual methods which can be time-consuming if results for multiple parameters are required. Using a solid-phase extraction procedure with an anion exchanger in this work, we have developed and optimized a multi-residue method for the extraction of 21 phenols and acids in sewage influent and effluent. The phenols and acids were then selectively eluted in separate fractions and were converted into pentafluoropropionyl (PFP) and tert butyldimethylsilyl (TBDMS) derivatives, respectively, for gas chromatography-mass spectrometric (GC/MS) determination. When applied to the sewage samples under study, the results for nonylphenol, bisphenol A (BPA), triclosan (TCS), 17ss estradiol (E2), estrone (E1), salicylic acid, ibuprofen, naproxen, diclofenac, and a few other acidic drugs were consistent with those determined previously by individual methods. Using the same procedure, we also report, for the first time, the occurrence of 2-phenylphenol and parabens in those sewage samples. PMID- 16257297 TI - Determination of atmospheric concentrations of inorganic anions by ion chromatography following ultrasonic extraction. AB - The capability of the suppressed conductometric detection ion chromatography (IC) was investigated for the separation and determination of inorganic anions (F-, Cl , NO3- and SO4(2-)) in standard reference materials SRM-1648 urban particulate matter following ultrasonic extraction. The effects of the cationic surfactant (SDS) and the anionic surfactant (CTAB) on ultrasonic extraction efficiency of inorganic anions from complex matrix of airborne particulate matter were investigated. The results showed that surfactant can enhance the extraction efficiency. Finally, the concentrations of inorganic anions in the atmosphere of the city of Isfahan were determined. The results showed a trend of SO4(2-) > NO3- > Cl- > F-. PMID- 16257299 TI - Organotin speciation in environmental matrices by automated on-line hydride generation-programmed temperature vaporization-capillary gas chromatography-mass spectrometry detection. AB - In the present contribution, a new automated on-line hydride generation methodology was developed for dibutyltin and tributyltin speciation at the trace level, using a programmable temperature-vaporizing inlet followed by capillary gas chromatography coupled to mass spectrometry in the selected ion-monitoring mode acquisition (PTV-GC/MS(SIM)). The methodology involves a sequence defined by two running methods, the first one configured for hydride generation with sodium tetrahydroborate as derivatising agent and the second configured for speciation purposes, using a conventional autosampler and data acquisition controlled by the instrument's software. From the method-development experiments, it had been established that injector configuration has a great effect on the speciation of the actual methodology, particularly, the initial inlet temperature (-20 degrees C; He: 150 ml/min), injection volume (2 microl) and solvent characteristics using the solvent venting mode. Under optimized conditions, a remarkable instrumental performance including very good precision (RSD < 4%), excellent linear dynamic range (up to 50 microg/ml) and limits of detection of 0.12 microg/ml and 9 ng/ml, were obtained for dibutyltin and tributyltin, respectively. The feasibility of the present methodology was validated through assays upon in-house spiked water (2 ng/ml) and a certified reference sediment matrix (Community Bureau of Reference, CRM 462, Nr. 330 dibutyltin: 68+/-12 ng/g; tributyltin: 54+/-15 ng/g on dry mass basis), using liquid-liquid extraction (LLE) and solid-phase extraction (SPE) sample enrichment and multiple injections (2 x 5 microl) for sensitivity enhancement. The methodology evidenced high reproducibility, is easy to work-up, sensitive and showed to be a suitable alternative to replace the currently dedicated analytical systems for organotin speciation in environmental matrices at the trace level. PMID- 16257300 TI - Low temperature bonding of poly(methylmethacrylate) electrophoresis microchips by in situ polymerisation. AB - A novel method for bonding poly(methyl methacrylate) (PMMA) electrophoresis microchips at the temperature below the glass transition temperature of PMMA based on in situ polymerization has been demonstrated. Methyl methacrylate (MMA) containing initiators was allowed to prepolymerize in an 85 degrees C water bath for 8 min and 15 min to produce a bonding solution and a dense molding solution, respectively. The channel plate of the PMMA microchip was fabricated by the UV initiated polymerization of the molding solution between a nickel template and a PMMA plate at room temperature. Prior to bonding, the blank cover was coated with a thin layer of the bonding solution and was bonded to the channel plate at 95 degrees C for 20 min under the pressure of binder clips. The attractive performance of the PMMA chips bonded by the new approach has been demonstrated by separating and detecting dopamine, catechol, three cations, and three organic acids in connection with end-column amperometric detection and contactless conductivity detection. PMID- 16257302 TI - Preparation of an extractant-impregnated porous membrane for the high-speed separation of a metal ion. AB - A novel impregnation method of extractants into a porous polymeric support is described. Bis(2-ethylhexyl)phosphate (HDEHP) was impregnated onto an n octadecylamino group of the polymer chain grafted onto the pore surface of a porous hollow-fiber membrane. First, an epoxy-group-containing polymer chain was appended onto the porous membrane by radiation-induced graft polymerization of glycidyl methacrylate (GMA). Second, n-octadecylamine was added to the graft chain via an epoxy-ring opening reaction to yield a hydrophobic group density of 3.0 mmol/g of the GMA-grafted fiber. Finally, HDEHP was impregnated to the n octadecylamino group. The amount of impregnated HDEHP of 2.1 mmol/g of the GMA grafted fiber was attained while retaining the liquid permeability of the porous membrane. An yttrium solution was forced to permeate through the pores of the HDEHP-impregnated porous hollow-fiber membrane. The higher permeation rate of the yttrium solution led to the higher adsorption rate of yttrium because of a negligible diffusional mass-transfer resistance. In addition, a high stability of impregnated HDEHP was observed after the repeated use of adsorption with 50 mg Y/L yttrium solution and elution with 7 M nitric acid. PMID- 16257301 TI - Expanded electrical model of a contactless conductivity detector: development and verification. AB - A theoretical model of the contactless conductivity detector (CCD) has been developed consisting of a network of resistors and capacitors. The output of the model is compared to experimental results and to the output of a simpler model. Experimentally, a lock-in amplifier is added to the detection scheme of the contactless conductivity detector to provide a more sensitive method of signal isolation. The detector is assembled on a printed circuit board with the electrodes in a co-axial configuration. The electrodes are chosen to allow for use with fused silica capillaries in capillary electrophoresis. The use of a lock in amplifier in place of a previous rectification/filtering circuit allows for an approximate 10-fold improvement in S/N. The detector shows a linear response to changes in excitation voltage and to changes in analyte concentration. Mass limits of detection of 60, 63, and 50 fg are determined for the inorganic cations potassium, sodium, and lithium, respectively (for a signal three times the level of the rms noise). PMID- 16257303 TI - Stability problems of polyether ether ketone and ethylene-tetrafluoroethylene copolymer tubing in simulated moving bed operation. AB - Polymeric capillaries made from polyether ether ketone (PEEK) or ethylene tetrafluoroethylene copolymer (Tefzel) are considered as highly inert and chemically resistant materials used as standard equipment in HPLC and simulated moving bed (SMB) applications. During several racemate separations using a SMB unit equipped with these tubes a formation of micro-holes was observed. All separations had in common that a high content of an alkane was used in the mobile phase. The patterns of damage and possible reasons causing the leakages of the capillaries are discussed. Polymeric tubing had to be replaced by stainless steel capillaries for the enantiomer separations in order to ensure safety of workers, GMP status of products and control leakages. PMID- 16257304 TI - High-performance liquid chromatography of governing liquid to detect illegal bovine milk's addition in water buffalo Mozzarella: comparison with results from raw milk and cheese matrix. AB - A method to detect fraudulent addition of bovine milk in water buffalo Mozzarella cheese by gradient high-performance liquid chromatography (RP-HPLC), relying on the measurement of quantity ratios within beta-lactoglobulin protein family, is described. Analyses were performed on raw milk, cheese matrix and cheese governing liquid using a C4 column and UV detection. This work demonstrated that bovine milk addition during cheesemaking can be detected in governing liquid of Mozzarella down to the EU law limit of 1% as well as in raw milk and cheese matrix. A significant lowering of peaks' areas and heights was observed in cheese matrix and governing liquid samples in comparison with the corresponding milk ones, possibly due to proteins' degradation during the cheesemaking process. The results show that, unlike previous works reported, the use of a matrix-specific calibration curve is essential in order to achieve a proper quantitation of beta lactoglobulin proteins, thus allowing a reliable estimation of bovine milk addition. PMID- 16257305 TI - Analysis of nicotinoid insecticides residues in honey by solid matrix partition clean-up and liquid chromatography-electrospray mass spectrometry. AB - An analytical method for the routine simultaneous determination of four nicotinoid insecticides (acetamiprid, imidacloprid, thiacloprid and thiamethoxam) in commercial multifloral honey was developed. Fortified honey samples, dissolved in water, were cleaned up through Extrelut NT20 column and, finally, insecticides were eluted with dichloromethane. The eluate was evaporated, the residue redissolved in methanol and then analyzed by LC-ESI(+)-MS. Average recoveries of the four analytes were in the range of 76% and 99% at both spiking levels 0.1 and 1.0 mg kg(-1). Relative standard deviations (RSDs) were less than 10% for all of the recovery tests. The detection limits (LODs) of the method ranged from 0.01 to 0.1 mg kg(-1) for the different insecticides studied. The developed method is linear over the range assayed, 0.5-5.0 microg mL(-1), with linear correlation coefficients higher than 0.9993. PMID- 16257306 TI - Comments to "Investigations of a new field in gas chromatography" by V.G. Berezkin, A.B. Lapin and J.B. Lipsky. PMID- 16257307 TI - Ophthalmic infections and their anti-infective challenges. AB - This introduction provides an overview of the succeeding articles contained within this supplement on the new fourth-generation fluoroquinolone antibiotic product, moxifloxacin ophthalmic solution 0.5% (VIGAMOX, Alcon Laboratories, Inc., Fort Worth, TX). Moxifloxacin was developed specifically to address the increasing incidence of resistance to earlier-generation antibiotic molecules. Structural modifications to the moxifloxacin molecule have decreased the likelihood of the development of resistant organisms. This antibiotic has been shown to possess greater activity than previous-generation molecules against gram positive bacteria while maintaining excellent potency against gram-negative organisms and nontuberculous (atypical) mycobacteria. Moxifloxacin ophthalmic solution 0.5% exhibits enhanced bioavailability due to a unique molecular structure that combines high lipophilicity for enhanced corneal penetration with high aqueous solubility at physiological pH. Numerous studies have shown that moxifloxacin ophthalmic solution 0.5% has high potency against a broad range of microbial species and a favorable profile in terms of safety and tolerability. The results presented in this supplement provide additional evidence for the potential benefits of moxifloxacin ophthalmic solution 0.5% in surgical prophylaxis and treatment of sight-threatening infections, such as bacterial conjunctivitis, endophthalmitis and keratitis. PMID- 16257308 TI - In vitro and in vivo potency of moxifloxacin and moxifloxacin ophthalmic solution 0.5%, a new topical fluoroquinolone. AB - Fluoroquinolones are a class of synthetic antibacterial agents that were approved for ocular therapy in 1991 and have become popular therapy for the treatment and prevention of various ocular infections. These agents are synthetic, broad spectrum, rapidly bactericidal, and have good penetration into ocular tissues. Their main mechanism of action is the inhibition of bacterial enzymes needed for bacterial DNA synthesis. However, antibiotic resistance occurred swiftly to the earlier fluoroquinolones and better fluoroquinolones were needed. The fourth generation fluoroquinolones, such as moxifloxacin and gatifloxacin, have enhanced activity against gram-positive bacteria while retaining potent activity against most gram-negative bacteria. These fourth-generation fluoroquinolones have improved penetration into the anterior chamber and have also demonstrated increased in vivo efficacy in several animal models of ocular infections. In addition, topical ophthalmic antibiotic products can deliver antibiotic concentrations directly to the eye that are thousands of times higher than their MICs. This article reviews published data describing the in vitro potency of moxifloxacin and its in vivo activity for treating and preventing experimental ocular infections. PMID- 16257309 TI - Ocular pharmacokinetics of moxifloxacin after topical treatment of animals and humans. AB - The ocular penetration and pharmacokinetics of moxifloxacin in comparison to other fluoroquinolones (ofloxacin, ciprofloxacin, gatifloxacin, norfloxacin, levofloxacin, and lomefloxacin) have been determined by in vitro and ex vivo techniques, as well as in animal and human studies. This article reviews the original pharmacokinetics work performed by Alcon and other studies reported in the ocular fluoroquinolone literature. The results consistently demonstrate higher maximum concentrations for moxifloxacin relative to the other fluoroquinolones in ocular tissues with levels well above its minimum inhibitory concentrations for relevant ocular pathogens. This superior performance is due to the unique structure of moxifloxacin that combines high lipophilicity for enhanced corneal penetration with high aqueous solubility at physiological pH. The latter property creates a high concentration gradient at the tear film/corneal epithelial interface providing a driving force for better ocular penetration for moxifloxacin. In addition, the higher concentration of moxifloxacin in VIGAMOX (i.e., 0.5% vs. 0.3%) allows more antibiotic to be available to ocular tissues. It is clear from the array of studies summarized in this report that moxifloxacin penetrates ocular tissues better (two- to three fold) than gatifloxacin, ciprofloxacin, ofloxacin, or levofloxacin. This consistent, enhanced penetration of topical moxifloxacin offers powerful advantages for ophthalmic therapy. PMID- 16257310 TI - Safety of moxifloxacin as shown in animal and in vitro studies. AB - Topical treatment of ocular bacterial infection is practiced widely, and the choice of the antibacterial agent depends on the nature of the infection, including the susceptibility of the organism, the tissue affected, and the safety profile of the agent. Moxifloxacin is a fourth-generation fluoroquinolone approved for ophthalmic use as moxifloxacin ophthalmic solution 0.5% (VIGAMOX, Alcon, Fort Worth, TX). Moxifloxacin ophthalmic solution 0.5% is self-preserved at a near-neutral pH of 6.8. In treating ocular infection, the three important aspects of therapeutic control are potency, penetration of the drug to the target site, and safety of the drug and the drug product. Moxifloxacin ophthalmic solution 0.5% provides antibacterial potency and high penetration of target ocular tissues. The ocular and systemic safety profile of moxifloxacin compares favorably with those of other fluoroquinolone antimicrobial agents, with a low risk of recognized quinolone-related toxicity. In vitro studies of fluoroquinolones with human or rabbit corneal epithelial cells or keratocytes suggest that moxifloxacin is similar in cytotoxicity potential to other drugs of this family. Specialized in vivo corneal wound-healing studies draw little distinction between moxifloxacin-treated eyes and those treated with other fluoroquinolones. Repeated-dose topical ocular studies in rabbits and monkeys, with high concentrations (up to 3%) of moxifloxacin and at treatment durations and regimens well in excess of label-prescribed use, demonstrated a high safety margin for ocular and extraocular tissues. Cornea, the tissue with highest exposure, was found to be unaffected by these high exposures, with slit-lamp biomicroscopy, corneal thickness measurement, intraocular pressure, and specular microscopy of the corneal endothelium (monkeys only), and histologic evaluation showing no effects, as compared with controls. Moxifloxacin ophthalmic solution 0.5% affords superior efficacy and ocular tissue penetration, with a favorable safety profile. PMID- 16257311 TI - Clinical safety of moxifloxacin ophthalmic solution 0.5% (VIGAMOX) in pediatric and nonpediatric patients with bacterial conjunctivitis. AB - Five independent, multicentered, double-masked, parallel, controlled studies were conducted to determine the safety of moxifloxacin ophthalmic solution 0.5% (VIGAMOX) in pediatric and nonpediatric patients with bacterial conjunctivitis. Patients were randomized into one of two treatment groups in each study and received either moxifloxacin ophthalmic solution 0.5% b.i.d. or t.i.d. or a comparator. A total of 1,978 patients (918 pediatric and 1,060 nonpediatric) was evaluable for safety. The most frequent adverse event in the overall safety population was transient ocular discomfort, occurring at an incidence of 2.8%, which was similar to that observed with the vehicle. No treatment-related changes in ocular signs or visual acuity were observed with moxifloxacin ophthalmic solution 0.5%, except for one clinically relevant change in visual acuity. Thus, based upon a review of adverse events and an assessment of ocular parameters, moxifloxacin ophthalmic solution 0.5% formulated without the preservative, benzalkonium chloride, is safe and well tolerated in pediatric (3 days-17 years of age) and nonpediatric (18-93 years) patients with bacterial conjunctivitis. PMID- 16257312 TI - Future of ophthalmic anti-infective therapy and the role of moxifloxacin ophthalmic solution 0.5% (VIGAMOX). AB - The vintage antibiotics that were available in the 1950s-1980s were sometimes toxic, had limited spectra, and were bacteriostatic agents, and they have been replaced by significantly broader-spectrum therapies. We ask more of our future antibiotic products for ophthalmology: they must be 1) broad spectrum, 2) convenient to use, 3) useful prophylactically, 4) effective therapeutically, 5) benzalkonium chloride-free, 6) comfortable, and 7) nontoxic. The emergence of antibiotic resistance has focused us on more potent agents effective against resistant strains of bacteria. Fluoroquinolones have become a dominant family of ophthalmic antibiotics. But even the older fluoroquinolones (e.g., ofloxacin, ciprofloxacin) have lost much of their effectiveness against some important ocular isolates. Considering all of the characteristics for an ideal ophthalmic antibiotic product available today, moxifloxacin ophthalmic solution 0.5% represents a primary antibiotic product of choice for treating and preventing ophthalmic infections. PMID- 16257313 TI - Overview of the potency of moxifloxacin ophthalmic solution 0.5% (VIGAMOX). AB - Antibiotics have been the mainstay of therapy for infectious diseases since their origins in the 1940s. As microorganisms changed and resistance developed, more advanced antibiotics were ultimately needed to provide adequate coverage and spectrum. By selecting optimal antibiotics and dosing regimens, clinicians can avoid treatment failures and adverse events and can help prevent the emergence of further antibiotic resistance. The fourth-generation ophthalmic fluoroquinolones include moxifloxacin (VIGAMOX, Alcon Laboratories, Inc., Fort Worth, TX) and gatifloxacin (Zymar, Allergan, Irvine, CA), and they are now approved for the treatment of bacterial conjunctivitis. This review highlights four scientific methods that compare and rank antibiotic potencies and predict their clinical efficacy and their propensity to develop resistance: 1) in vitro assay for minimum inhibitory concentrations, 2) in vivo models for pharmacokinetic and pharamacodynamic properties, 3) therapeutic index or inhibitory quotient, and 4) in vitro assay for mutant prevention concentration. The fourth-generation ophthalmic fluoroquinolones perform well in these assays. Both antibiotics have better in vitro activity against gram-positive bacteria than ciprofloxacin or ofloxacin. Moxifloxacin penetrates better into ocular tissues than gatifloxacin and older fluoroquinolones; in vitro activity of moxifloxacin and gatifloxacin against gram-negative bacteria is similar to that of older fluoroquinolones. Moxifloxacin also has better mutant prevention characteristics than other fluoroquinolones. These findings support the use of the newer fluoroquinolones for the prevention and treatment of serious ophthalmic infections (e.g., keratitis, endophthalmitis) caused by susceptible bacteria. PMID- 16257314 TI - Enhanced chemical oxidation of aromatic hydrocarbons in soil systems. AB - Fenton's destruction of benzene, toluene, ethylbenzene, and xylene (BTEX) was investigated in soil slurry batch reactors. The purpose of the investigation was to quantify the enhancement of oxidation rates and efficiency by varying process conditions such as iron catalyst (Fe(II) or Fe(III); 2, 5, and 10mM), hydrogen peroxide (H2O2; 30, 150, 300 mM), and metal chelating agents (l-ascorbic acid, gallic acid, or N-(2-hydroxyethyl)iminodiacetic acid). Rapid contaminant mass destruction (97% after 3h) occurred in the presence of 300 mM H2O2 and 10 mM Fe(III). An enhanced removal rate (>90% removal after 15 min and 95% removal after 3h) was also observed by combining Fe(III), N-(2-hydroxyethyl)iminodiacetic acid and 300 mM H2O2. The observed BTEX mass removal rate constants (3.6-7.8 x 10(-4)s(-1)) were compared to the estimated rate constants (4.1-10.1 x 10(-3)s( 1)). The influence of non-specific oxidants loss (by reaction with iron hydroxides and soil organic matter) was also explored. PMID- 16257315 TI - Effects of in-situ ozonation on indigenous microorganisms in diesel contaminated soil: survival and regrowth. AB - Soil column experiments were conducted to investigate the effects of chemical oxidation on the survival of indigenous microbes (i.e., heterotrophic microbes, phenanthrene-degrading microbes, and alkane-degrading microbes) for field soil contaminated with diesel fuel. Rapid decreases of total petroleum hydrocarbons (TPH) and aromatics of diesel fuel were observed within the first 60 min of ozone injection; after 60 min, TPH and aromatics decreased asymptotically with ozonation time. The three types of indigenous microbes treated were very sensitive to ozone in the soil column experiment, hence the microbial population decreased exponentially with ozonation time. The numbers of heterotrophic, alkane degrading, and phenanthrene-degrading bacteria were reduced from 10(8) to 10(4), 10(7) to 10(3), and 10(6) CFU g soil(-1) to below detection limit after 900 min of ozonation, respectively. Except for the soil sample ozonated for 900 min, incubation of ozone-treated soil samples that were not limited by oxygen diffusion showed further removal of TPH. The soil samples that were ozonated for 180 min exhibited the lowest concentration of TPH and the highest regrowth rate of the heterotrophic and alkane-degrading populations after the 9 weeks of incubation. PMID- 16257316 TI - An evaluation of reactive filter media for treating landfill leachate. AB - A laboratory bench-scale column study was conducted to evaluate permeable reactive filter materials as a new method for removal of heavy metals and inorganic nitrogen from landfill leachate. Mixtures of sand and peat, blast furnace slag (BFS) and peat, and Polonite and peat were tested by loading columns with leachate collected from a pond at Tvetaverket Landfill, Sweden. Sand, peat and Polonite represent natural materials. BFS is a by-product from steel-works. The metal treatment efficiencies of the media were assessed and Polonite was found to perform best, where Mn, Fe, Zn and Cu concentrations were removed by 99%, 93%, 86% and 67%, respectively. This material was also able to reduce inorganic N by 18%. The BFS showed good removal efficiency for Cu (66%), Zn (62%), Ni (19%) and Mo (16%). The sand-peat mixture did not demonstrate a promising removal capacity for any of the elements studied with the exception of Cu (25%). The removal of different elements was suggested to be a combination of several factors, i.e. precipitation, ion exchange and adsorption. Prior to full scale application of reactive filters at a landfill site, matrix selection, filter design and operational procedures must be developed. PMID- 16257318 TI - Characterization of carbons from olive cake by sorption of wastewater pollutants. AB - Studies has been conducted to compare the sorption properties between raw carbons made from olive cake and commercial activated carbons to remove aquatic pollutant such as heavy metal (HM), phenol (Ph), dodecylbenzenesulfonic acid-sodium salt detergent (DBSNa) and methylene blue dye (MB). Effect of acidic treatments by H2SO4, HCl and HNO3 on the sorption properties of olive cake carbon (OCC) were studied by mass titration, SEM photographs, sorption isotherms. It is found that acidic treatment changes the surface properties of OCC but do not enhance its sorption capacity. Compared to commercial activated carbons the OCC derivatives generally are equally able to uptake HM and Ph from solution but MB and DBSNa are not. This different behaviour is to attribute to manufacturing and activation treatments so as surface groups of the precursor sorbent material. For the heavy metals, chromium and silver were removed effectively but to small extent cadmium. This may be because of the various charge densities of metal elements tested. The results of the multiple experiments indicate that sorption of cadmium ions can be significantly improved by the presence of complexing agents sorbed from the carbon. An empirical mathematical form is proposed to correlate experimental data and to compare the performance of the different sorbent materials. PMID- 16257319 TI - Thermal degradation of fluorotelomer treated articles and related materials. AB - This study reports the first known studies to investigate the thermal degradation of a polyester/cellulose fabric substrate ("article") treated with a fluorotelomer-based acrylic polymer under laboratory conditions conservatively representing typical combustion conditions of time, temperature, and excess air level in a municipal incinerator, with an average temperature of 1000 degrees C or greater over approximately 2s residence time. The results demonstrate that the polyester/cellulose fabric treated with a fluorotelomer-based acrylic polymer is destroyed and no detectable amount of perfluorooctanoic acid (PFOA) is formed under typical municipal incineration conditions. Therefore, textiles and paper treated with such a fluorotelomer-based acrylic polymer disposed of in municipal waste and incinerated are expected to be destroyed and not be a significant source of PFOA in the environment. PMID- 16257317 TI - Multi-component behavior of fixed-bed adsorption of dioxins by activated carbon fiber. AB - The multi-component behavior of fixed-bed adsorption of dioxins (DXNs) was examined through detailed analyses of the concentration profiles of isomers in fixed-bed activated carbon fiber (ACF). Regularities in both adsorption rates and strengths were clarified. (1) The rate of transfer in the adsorption of polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans (PCCDs/DFs) tends to increase with decreasing number of chlorine substituents. Axial dispersion also tends to increase with a decreasing number of chlorine substituents under our experimental conditions. (2) Homologues with the same number of chlorine substituents in PCDDs/DFs have similar adsorption strengths. The adsorption strength of PCDD/DF isomers is probably greater than that of co planar polychlorinated biphenyls (co-PCB) isomers when the number of chlorine substituents is identical. (3) The adsorption strength of isomers depends on their molecular structure. In PCDDs/DFs the toxic isomers, all of which have vicinal chlorine substituents at the 2, 3, 7 and 8 positions, are relatively strong. It is clear, especially in TeCDDs, that isomers with vicinal chlorine substituents are stronger than isomers without. In co-PCBs, isomers without chlorine substituents at ortho positions are stronger than those with, and (4) A close analogy exists between the adsorption strength order for ACF and the elution order in gas chromatography (GC). PMID- 16257320 TI - Reutilization of granite powder as an amendment and fertilizer for acid soils. AB - The properties of granite powders--a granite manufacturing waste product-were analyzed to assess their potential use as amendments and fertilizers on acid soils. Two types of powders were characterized: one produced during cutting of granite with a diamond-edged disc saw, comprising only rock powder, the other produced during cutting with a multi-blade bandsaw, containing calcium hydroxide and metal filings added during the cutting procedure. The acid neutralizing capacity of the granite powders was assessed in short- (2-3 h) and medium-term (1 30 d) experiments. The powders showed a buffering capacity at around pH 8, which corresponded to the rapid dissolution of basic cations, and another buffering effect at pH<4.5, attributable to the dissolution of Fe and Al. The acid neutralizing capacity (ANC) determined in the short-term experiments, to a final pH of 4.5, varied between 5 and 61 cmol H+kg(-1) powder. The ANC to pH 4.5 obtained in the medium-term experiments was much higher than that obtained in the short-term experiments, reaching a maximum ANC value of 200 cmol H+kg(-1) powder. There was no great difference in the neutralizing capacity determined at between 1 and 30 d. The most abundant elements in acid solutions obtained at the end of medium-term experiments were Mg and Ca for disc saw powders, whereas Ca and Fe (at pH<5) were the most soluble elements in the bandsaw powders. The rapid release of these cations suggests the possible effective use of the granite powders as a source of nutrients on being added to acid soils. PMID- 16257321 TI - Influence of ozonation on extractability of Pb and Zn from contaminated soils. AB - The effect of soil ozonation on Pb and Zn extraction with EDTA, bioavailability (Ruby's Physiologically Based Extraction Test, PBET) and mobility (Toxicity Characteristic Leaching Procedure, TCLP) of Pb was studied on contaminated soils taken from 7 different locations in the Mezica Valley, Slovenia. EDTA extraction (40 mmol kg(-1)) removed from 27.4+/-1.5% to 64.8+/-1.5% of Pb, and from 1.9+/ 0.2% to 22.4+/-2.0% of Zn from tested soils, and significantly reduced soil Pb bioavailability (PBET) and mobility (TCLP). Pretreatment of tested soils with ozone before EDTA extraction enhanced EDTA extractability of Pb for 11.0 to 28.9%, but had no effect on the extractability of Zn. In most of the soils, ozonation had no statistically significant effect on bioavailability and mobility of Pb, residual after EDTA extraction. Using linear regression analysis we found a significant increase (p<0.01) in EDTA extractability of Pb after soil ozonation in soils with a higher initial Pb content. EDTA extractability of Pb after soil ozonation was also significantly higher for soils with a lower Pb extractability when treated with EDTA alone. We found no correlation between soil organic matter content and the percentage of the Pb fraction bound to soil organic matter (where from 25.6+/-1.3% to 73.2+/-0.6% of Pb reside in tested soils) and Pb extractability with EDTA after soil ozonation. PMID- 16257322 TI - Use of pyrite for pH control during hydrogenotrophic denitrification using metallic iron as the ultimate electron donor. AB - Maintenance of stable pH through provision of adequate buffering is of importance to many pollutant removal processes where either acid or base is produced as a reaction product. The objective of this study was to evaluate the suitability of pyrite (FeS2) as an in situ buffering agent for arresting pH increase during metallic iron assisted hydrogenotrophic denitrification. Pyrite is considered promising for this purpose because it is a mineral which is unstable under moderately reducing, i.e., anoxic conditions, where such denitrification takes place, and therefore expected to consume hydroxide ions produced due to hydrogenotrophic denitrification reactions and get oxidized to ferrous hydroxide Fe(OH)2. The theoretical basis for this buffering action was established through chemical speciation studies using the chemical speciation software, MINEQL+. Experimental evaluation of the buffering efficiency of pyrite showed that it was effective in arresting pH increase associated with denitrification in both batch systems and during flow through reactive porous media. Further, addition of pyrite had no demonstrable toxic effect on the denitrifying microorganisms, though elevated sulfate concentration was seen in the effluent after denitrification. PMID- 16257323 TI - Microbial activity in a combined UASB-activated sludge reactor system. AB - A combined upflow anaerobic sludge bed-activated sludge (UASB-AS) reactor system with consistently wasting of excess biomass was used to treat suspended-solids pre-settled piggery wastewater (COD=2000 mg l(-1), total Kjeldahl nitrogen TKN=400 mg l(-1), suspended solids=250-400 mg l(-1)). Thus, the activity of nitrogen-related microbial groups in each individual bioreactor was investigated. When the granules retention time (GRT) of 20-50 d in the UASB reactor, the solids retention time (SRT) of 10-25 d in the AS reactor and the recycle-to-influent ratio (Re) of 1 were maintained, the combined system removed 95-97% of chemical oxygen demand (COD), 100% of TKN and 54-55% of total nitrogen (TN). Denitrification and methanogenesis occurred in the UASB reactor so that both biochemical processes contributed to most of the COD removal and, complete nitrification (most of the TKN removal) occurred in the AS reactor. Compact granules with good settling abilities developed in the UASB reactor, and rapid rates of granulation of break-up granules in the UASB reactor were confirmed by experiments. The activity of nitrifiers and denitrifiers (an=0.68-0.87; adn=0.55 0.70) and the calculated specific nitrification and denitrification rates (qn=0.26-0.47 mg NH4+ -N mg VSS(-1)d(-1); qdn=0.046-0.076 mg NOx- -N mg VSS(-1)d( 1)) significantly increased with decreasing SRT and GRT, respectively. Accordingly, the combined UASB-AS reactor system should be regarded a promising alternative for the removal of organic carbon and nitrogen from piggery wastewater. PMID- 16257324 TI - Efforts to address gender inequalities must begin at home. PMID- 16257325 TI - Who is Harriet Miers? PMID- 16257326 TI - The trials of cancer chemoprevention. PMID- 16257327 TI - Molecular targeted therapy for lung cancer. PMID- 16257328 TI - Management of severe head injury: can we do better? PMID- 16257329 TI - The end of beta blockers for uncomplicated hypertension? PMID- 16257330 TI - MDGs: chronic diseases are not on the agenda. PMID- 16257331 TI - The neglected epidemic of chronic disease. PMID- 16257332 TI - Oliver Brustle: fighting for stem cell research in Germany. PMID- 16257333 TI - Cortical stability of the femoral neck and hip fracture risk. PMID- 16257334 TI - Cortical stability of the femoral neck and hip fracture risk. PMID- 16257336 TI - Cortical stability of the femoral neck and hip fracture risk. PMID- 16257337 TI - More confusion. PMID- 16257338 TI - One-dose carboplatin in seminoma. PMID- 16257339 TI - Gefitinib plus best supportive care in previously treated patients with refractory advanced non-small-cell lung cancer: results from a randomised, placebo-controlled, multicentre study (Iressa Survival Evaluation in Lung Cancer). AB - BACKGROUND: This placebo-controlled phase III study investigated the effect on survival of gefitinib as second-line or third-line treatment for patients with locally advanced or metastatic non-small-cell lung cancer. METHODS: 1692 patients who were refractory to or intolerant of their latest chemotherapy regimen were randomly assigned in a ratio of two to one either gefitinib (250 mg/day) or placebo, plus best supportive care. The primary endpoint was survival in the overall population of patients and those with adenocarcinoma. The primary analysis of the population for survival was by intention to treat. This study has been submitted for registration with ClinicalTrials.gov, number 1839IL/709. FINDINGS: 1129 patients were assigned gefitinib and 563 placebo. At median follow up of 7.2 months, median survival did not differ significantly between the groups in the overall population (5.6 months for gefitinib and 5.1 months for placebo; hazard ratio 0.89 [95% CI 0.77-1.02], p=0.087) or among the 812 patients with adenocarcinoma (6.3 months vs 5.4 months; 0.84 [0.68-1.03], p=0.089). Preplanned subgroup analyses showed significantly longer survival in the gefitinib group than the placebo group for never-smokers (n=375; 0.67 [0.49-0.92], p=0.012; median survival 8.9 vs 6.1 months) and patients of Asian origin (n=342; 0.66 [0.48-0.91], p=0.01; median survival 9.5 vs 5.5 months). Gefitinib was well tolerated, as in previous studies. INTERPRETATION: Treatment with gefitinib was not associated with significant improvement in survival in either coprimary population. There was pronounced heterogeneity in survival outcomes between groups of patients, with some evidence of benefit among never-smokers and patients of Asian origin. PMID- 16257340 TI - Trends in head injury outcome from 1989 to 2003 and the effect of neurosurgical care: an observational study. AB - BACKGROUND: Case fatality rates after all types of blunt injury have not improved since 1994 in England and Wales, possibly because not all patients with severe head injury are treated in a neurosurgical centre. Our aims were to investigate the case fatality trends in major trauma patients with and without head injury, and to establish the effect of neurosurgical care on mortality after severe head injury. METHODS: We analysed prospectively collected data from the Trauma Audit and Research Network database for patients presenting between 1989 and 2003. Mortality and odds of death adjusted for case mix were compared for patients with and without head injury, and for those treated in a neurosurgical versus a non neurosurgical centre. FINDINGS: Patients with head injury (n=22,216) had a ten fold higher mortality and showed less improvement in the adjusted odds of death since 1989 than did patients without head injury (n=154,231). 2305 (33%) of patients with severe head injury (presenting between 1996 and 2003) were treated only in non-neurosurgical centres; such treatment was associated with a 26% increase in mortality and a 2.15-fold increase (95% CI 1.77-2.60) in the odds of death adjusted for case mix compared with patients treated at a neurosurgical centre. INTERPRETATION: Since 1989 trauma system changes in England and Wales have delivered greater benefit to patients without head injury. Our data lend support to current guidelines, suggesting that treatment in a neurosurgical centre represents an important strategy in the management of severe head injury. PMID- 16257341 TI - Should beta blockers remain first choice in the treatment of primary hypertension? A meta-analysis. AB - BACKGROUND: Beta blockers have been used widely in the treatment of hypertension and are recommended as first-line drugs in hypertension guidelines. However, a preliminary analysis has shown that atenolol is not very effective in hypertension. We aim to substantially enlarge the data on atenolol and analyse the effect of different beta blockers. METHODS: The Cochrane Library and PubMed were searched for beta blocker treatment in patients with primary hypertension. Data were then entered into the Cochrane Collaboration Review Manager package and were summarised in meta-analyses. 13 randomised controlled trials (n=105 951) were included in a meta-analysis comparing treatment with beta blockers with other antihypertensive drugs. Seven studies (n=27 433) were included in a comparison of beta blockers and placebo or no treatment. FINDINGS: The relative risk of stroke was 16% higher for beta blockers (95% CI 4-30%) than for other drugs. There was no difference for myocardial infarction. When the effect of beta blockers was compared with that of placebo or no treatment, the relative risk of stroke was reduced by 19% for all beta blockers (7-29%), about half that expected from previous hypertension trials. There was no difference for myocardial infarction or mortality. INTERPRETATION: In comparison with other antihypertensive drugs, the effect of beta blockers is less than optimum, with a raised risk of stroke. Hence, we believe that beta blockers should not remain first choice in the treatment of primary hypertension and should not be used as reference drugs in future randomised controlled trials of hypertension. PMID- 16257342 TI - Interaction between CHEK2*1100delC and other low-penetrance breast-cancer susceptibility genes: a familial study. AB - BACKGROUND: The allele CHEK2*1100delC doubles the risk of breast cancer in unselected women, but could confer a greater risk in women with a family history of the disease, particularly of bilateral breast cancer. Our aim was to measure the risk of breast cancer in relatives of women with bilateral breast cancer who were carriers of this allele. METHODS: A population-based series of 469 bilateral breast cancer cases ascertained through English cancer registries were genotyped for CHEK2*1100delC. Standardised incidence ratios (SIRs) and cumulative risks were calculated for breast cancer, prostate cancer, and all other cancers in the first-degree relatives of carriers and non-carriers. FINDINGS: The relatives of bilateral cases who were wild-type for CHEK2 had three times the population risk of female breast cancer (145 cases: SIR 3.48 (95% CI 2.96-4.09), twice the risk of prostate cancer (34 cases: SIR 2.41, 1.67-3.36) and a large excess of male breast cancer (five cases: SIR 15.06, 4.92-35.36). Relatives of those who were carriers of CHEK2*1100delC had a substantially higher risk of breast cancer (eight cases: SIR 12.11, 5.23-23.88) and possibly prostate cancer (two cases: SIR 9.87, 1.20-35.67). INTERPRETATION: These data suggest a multiplicative interaction between CHEK2*1100delC and other unknown susceptibility genes. In women with a family history of bilateral disease, CHEK2*1100delC confers a high lifetime risk and might be useful for predictive testing. Bilateral breast cancer cases and their families are likely to provide an efficient basis for identification of additional low-penetrance breast-cancer genes. PMID- 16257343 TI - Effect of cruciferous vegetables on lung cancer in patients stratified by genetic status: a mendelian randomisation approach. AB - Whether consumption of cruciferous vegetables protects against lung cancer is unclear, largely because of potential confounding factors. We therefore studied the role of cruciferous vegetables in lung cancer after stratifying by GSTM1 and GSTT1 status, two genes implicated in the elimination of isothiocyanates, the likely chemopreventative compound. In 2141 cases and 2168 controls, weekly consumption of cruciferous vegetables protected against lung cancer in those who were GSTM1 null (odds ratio=0.67, 95% CI 0.49-0.91), GSTT1 null (0.63, 0.37 1.07), or both (0.28, 0.11-0.67). No protective effect was seen in people who were both GSTM1 and GSTT1 positive (0.88, 0.65-1.21). Similar protective results were noted for consumption of cabbage and a combination of broccoli and brussels sprouts. These data provide strong evidence for a substantial protective effect of cruciferous vegetable consumption on lung cancer. PMID- 16257344 TI - Advances in leishmaniasis. AB - Governed by parasite and host factors and immunoinflammatory responses, the clinical spectrum of leishmaniasis encompasses subclinical (inapparent), localised (skin lesions), and disseminated infection (cutaneous, mucosal, or visceral). Symptomatic disease is subacute or chronic and diverse in presentation and outcome. Clinical characteristics vary further by endemic region. Despite T cell-dependent immune responses, which produce asymptomatic and self-healing infection, or appropriate treatment, intracellular infection is probably life long since targeted cells (tissue macrophages) allow residual parasites to persist. There is an epidemic of cutaneous leishmaniasis in Afghanistan and Pakistan and of visceral infection in India and Sudan. Diagnosis relies on visualising parasites in tissue or serology; culture and detection of parasite DNA are useful in the laboratory. Pentavalent antimony is the conventional treatment; however, resistance of visceral infection in India has spawned new treatment approaches--amphotericin B and its lipid formulations, injectable paromomycin, and oral miltefosine. Despite tangible advances in diagnosis, treatment, and basic scientific research, leishmaniasis is embedded in poverty and neglected. Current obstacles to realistic prevention and proper management include inadequate vector (sandfly) control, no vaccine, and insufficient access to or impetus for developing affordable new drugs. PMID- 16257345 TI - Preventing chronic diseases: how many lives can we save? AB - 35 million people will die in 2005 from heart disease, stroke, cancer, and other chronic diseases. Only 20% of these deaths will be in high-income countries- while 80% will occur in low-income and middle-income countries. The death rates from these potentially preventable diseases are higher in low-income and middle income countries than in high-income countries, especially among adults aged 30 69 years. The impact on men and women is similar. We propose a new goal for reducing deaths from chronic disease to focus prevention and control efforts among those concerned about international health. This goal-to reduce chronic disease death rates by an additional 2% annually--would avert 36 million deaths by 2015. An additional benefit will be a gain of about 500 million years of life over the 10 years from 2006 to 2015. Most of these averted deaths and life-years gained will be in low-income and middle-income countries, and just under half will be in people younger than 70 years. We base the global goal on worldwide projections of deaths by cause for 2005 and 2015. The data are presented for the world, selected countries, and World Bank income groups. PMID- 16257346 TI - Eye know it's red. PMID- 16257348 TI - Tryptophan hydroxylase polymorphism is associated with age of onset of alcoholism related behaviors. AB - The purpose of this study was to explore the association of a tryptophan hydroxylase gene polymorphism (TPH1 A218C) with the age of alcoholism onset in a Korean population. The genotype and allele frequencies of TPH1 were investigated in 182 male hospitalized patients who met Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition, Text Revision (DSM-IV-TR) criteria for alcohol dependence. Alcoholics with the TPH1 AA or AC genotypes had an earlier age of disease onset (median age of onset, 26.5 years) than those with the CC genotype (median age of onset, 30 years; p=.002). Age of onset has been used in classifying alcoholics. The TPH1 polymorphism may explain, in part, the biological basis for these typologies. PMID- 16257347 TI - Effectiveness of posthumous molecular diagnosis from a kept baby tooth. PMID- 16257349 TI - Effect of ethanol administration on Mg2+ transport across liver plasma membrane. AB - Acute and chronic ethanol administration results in a decrease in cellular Mg2+ content and an alteration of Mg2+ transport in liver cells. In this study we investigated the extent to which ethanol affects the Mg2+ transport mechanisms in the liver cell membrane. The functionality of these transport mechanisms was assessed in plasma membrane vesicles purified from livers acutely perfused with varying concentrations of alcohol, livers of animals fed with 6% ethanol for 3 weeks, and the respective controls. Acute alcohol administration had little or no effect on the Mg2+ extrusion mechanisms present in the apical and basolateral domains of the hepatocyte but completely impaired the Mg2+ entry mechanism present in the basolateral side of the cell. This effect was already evident at a dose of alcohol as small as 0.01% (approximately 1.5 mM). The chronic administration of ethanol, instead, impaired all the Mg2+ transport mechanisms irrespective of the location and directionality in a time-dependent manner. Taken together, these data indicate a selective sensitivity of the Mg2+ entry mechanism to acute alcohol administration, whereas the Mg2+ extrusion mechanisms are affected only after prolonged exposure to alcohol. These results suggest that the defect in hormone-activated Mg(2+) transport observed in the chronic EtOH model [Young, A., Cefaratti, C., & Romani, A. (2003). Chronic EtOH administration alters liver Mg2+ homeostasis. Am J Physiol 284, G57-G67] depends not only on a reduced cellular Mg2+ content but also on the impaired Mg2+ transport mechanisms present in the hepatocyte plasma membrane, in particular the Mg2+ entry pathway, which prevents the liver cell from restoring cellular Mg2+ homeostasis. PMID- 16257350 TI - Alcohol intake is associated with altered pulmonary function. AB - Little is known about the effect of moderate alcohol intake on lung function in the general population. Because moderate alcohol intake appears to reduce cardiovascular disease risk, we hypothesized that moderate alcohol intake is associated with better pulmonary function. To test this hypothesis, we examined the association between alcohol intake and pulmonary function, measured by spirometry, in a representative sample of U.S. adults who participated in the Third National Health and Nutrition Examination Survey. A stratified multistage clustered probability design was used to select a population-based sample. Data analyzed included alcohol intake, smoking status, education, body mass, sex, age, race, diabetes status, and CHF status. The Third National Health and Nutrition Examination Survey was conducted from 1988 to 1994 by the National Center for Health Statistics of the Centers for Disease Control and Prevention, Atlanta, GA. We analyzed data from 15,294 study participants who completed extensive questionnaires in the household and a comprehensive physical examination, including pulmonary function testing, either in the household or at a specially equipped mobile examination center. Low-to-moderate alcohol intake was not associated with reduced odds of obstructive lung function. In fact, increased odds for obstructive lung pattern were observed only in former heavy drinkers. In contrast, low-to-moderate alcohol intake was associated with better forced vital capacity and forced exhaled volume in 1s in the absence of obstruction, consistent with reduced odds for lung restriction. Using a logistic regression model, we found that individuals reporting alcohol consumption had a lower risk of lung restriction both before and after adjusting for confounding factors including smoking (P< or =.001). Alcohol intake-related reduced risk for restriction was associated with lower risk of CHF, diabetes, obesity, and lower markers of inflammation (white blood cell, fibrinogen, and C-reactive protein) consistent with less lung congestion, external restriction, and/or lung inflammation. Our analyses indicate that alcohol consumption, even at very modest intake levels, is associated with less lung restriction. PMID- 16257351 TI - Malondialdehyde-acetaldehyde adducts decrease bronchial epithelial wound repair. AB - Most people who abuse alcohol are cigarette smokers. Previously, we have shown that malondialdehyde, an inflammation product of lipid peroxidation, and acetaldehyde, a component of both ethanol metabolism and cigarette smoke, form protein adducts that stimulate protein kinase C (PKC) activation in bronchial epithelial cells. We have also shown that PKC can regulate bronchial epithelial cell wound repair. We hypothesize that bovine serum albumin adducted with malondialdehyde and acetaldehyde (BSA-MAA) decreases bronchial epithelial cell wound repair via binding to scavenger receptors on bronchial epithelial cells. To test this, confluent monolayers of bovine bronchial epithelial cells were grown in serum-free media prior to wounding the cells. Bronchial epithelial cell wound closure was inhibited in a dose-dependent manner (up to 60%) in the presence of BSA-MAA than in media treated cells (Laboratory of Human Carcinogenesis [LHC]-9 Roswell Park Memorial Institute [RPMI]). The specific scavenger receptor ligand, fucoidan, also stimulated PKC activation and decreased wound repair. Pretreatment with fucoidan blocked malondialdehyde-acetaldehyde binding to bronchial epithelial cells. When bronchial epithelial cells were preincubated with a PKC alpha inhibitor, Go 6976, the inhibition of wound closure by fucoidan and BSA-MAA was blocked. Western blot demonstrated the presence of several scavenger receptors on bronchial epithelial cell membranes, including SRA, SRBI, SRBII, and CD36. Scavenger receptor-mediated activation of PKC alpha may function to reduce wound healing under conditions of alcohol and cigarette smoke exposure where malondialdehyde-acetaldehyde adducts may be present. PMID- 16257352 TI - Regulation of alcohol intake with advancing age. AB - Previous surveys of alcohol use in the general population have not gathered sufficient data to allow for estimations of the blood alcohol levels (BACs) routinely achieved in survey participants. Our goal was to assess the influence of age on the estimated peak BAC achieved on typical drinking occasions in a representative sample (n=2,626) of the U.S. adult population. Variables related to the quantity and duration of alcohol consumption on typical drinking occasions were assessed by computer-assisted telephone interview. In addition, the height, weight, age, and gender of subjects were ascertained to be used in equations to predict the volume of distribution of ethanol (total body water). Prediction equations were used to estimate the probable peak BACs achieved during the typical drinking occasion. The survey identified 1,833 subjects ("current drinkers") of 18-89 years, who reported alcohol consumption within the past 12 months. Linear regression analyses performed on data from these "current drinkers" revealed that, for both men and women, there was an age-related decrease in the predicted peak BAC achieved on typical drinking occasions. The approaches used to modify the BAC with advancing age differed slightly for men and women, but both relied heavily upon a reduction in the quantity of consumption. PMID- 16257353 TI - Alcohol consumption and serum hormone levels during pregnancy. AB - Factors that change sex hormone levels during pregnancy may have long-term health consequences for the offspring, including changes in breast cancer risk. A cross sectional analysis of alcohol consumption and hormone levels in 339 pregnant women sampled from the Child Health and Development Study cohort was undertaken. Alcohol intake was queried from 1959 to 1966, long before any hazards of drinking during pregnancy were publicized. Third trimester serum hormone levels including estradiol and testosterone were analyzed. Among 339 pregnant women, 196 reported some alcohol consumption during pregnancy. The drinkers were divided into three groups with intake levels of 0.2-0.5, 0.6-2.0, and 2.1-12.5 ounces of ethanol per week. The second group corresponds to a median intake of approximately 2 drinks per week, and the last group corresponds to a median intake of approximately 1 drink per day, which is considered "light" to "moderate" drinking. Maternal estradiol levels were not associated with alcohol intake during pregnancy. However, serum testosterone was significantly lower, by 12.2%, in the latter two groups of drinking pregnant women, [confidence interval (CI)=-3.0 to 25.2] and 25.6% (CI=9.2-39.5), respectively. The alcohol intakes reported are far below those shown to cause fetal alcohol syndrome, or any of the fetal alcohol effects so far studied. Light alcohol intake during pregnancy is associated with lower maternal testosterone. The health implications are uncertain, but may include an increased breast density in the daughters of drinking mothers. PMID- 16257354 TI - Different stages in the temporal course of estrogen treatment produce opposite effects on voluntary alcohol consumption in male rats. AB - The purpose of this study was to examine alcohol consumption in different stages of the estrogen treatment. Three groups of castrated male Wistar rats were used. One group was treated with 5 microg of estradiol benzoate (E) per day per rat for 6 days and oil from days 7 to 12 (EO group). The second group was treated with oil for 6 days and E from days 7 to 12 (OE) and the third with E for 12 consecutive days (EE). The three groups were exposed to a choice of both water and ethanol (10%) before treatment (PreT), from days 7 to 12 of the oil or the E treatment (T2), and during 6 additional days in the post-treatment period (PosT). Alcohol was not available from days 1 to 6 of the oil or the E treatment (T1). Alcohol consumption in the EO group during T2 was higher than in PreT and PosT periods and all periods in the other two groups. In contrast, alcohol consumption during T2 was significantly lower than during the PreT of the OE group and T2 of the EE group. At the same time in the EE group, alcohol intake in the T2 was higher than in the PreT and the PosT periods. These results reveal the opposite effects of estrogen treatment on alcohol consumption, which apparently depended on the physiological conditions produced by the temporal course of hormone treatment. PMID- 16257355 TI - Anterior mandibular apical base augmentation in the surgical orthodontic treatment of mandibular retrusion. AB - The authors describe a surgical technique alternative to traditional pre-surgical orthodontics in order to increase the apical base in mandibular retrusion (class II, division I). This subapical osteotomy, optimizing inferior incisal axis without dental extractions and a long orthodontic treatment, associated to genioplasty permits to obtain an ideal labio-dento-mental morphology. This procedure avoids in some cases the need of a mandibular advancement and, if necessary, it reduces his entity with obvious advantages. PMID- 16257356 TI - Titanium screw implantation to the articular eminence for the treatment of chronic recurrent dislocation of the temporomandibular joint. AB - In this report, the technical details of a new surgical technique are presented for the treatment of chronic recurrent dislocation of the temporomandibular joint. The procedure is performed under local anesthesia. Two millimeters of skin incision is carried out 2-2.5cm below the articular eminence, and a tunnel is created above the superficial musculoaponeurotic plane toward the eminence. Then via a specially designed trocar, a 20mm titanium screw is implanted into the most prominent part of the eminence perpendicularly. Thereafter, the patient is told to actively open his/her mouth, and the luxation and position of the screw is checked with the C-arm fluoroscopy. Once the correct position is assured, the trocar is removed and the incision is sutured with 6/0 prolene. PMID- 16257357 TI - Effective density and mass attenuation coefficient for building material in Brazil. AB - This paper presents values for density and mass attenuation coefficient of building materials commonly used in Brazil. Transmission measurements were performed to provide input information for simulations with MCNP4B code. The structure for the clay bricks was simulated as a mix of all material layers and an effective density determined. The mass attenuation coefficients were determined for the 50-3,000 keV gamma-ray energy range. A comparison with results for similar materials found in the literature showed good agreement. PMID- 16257358 TI - Comment on Caprolactam study. PMID- 16257359 TI - Familial aggregation of airway responsiveness: a community-based study. AB - PURPOSE: We investigated the familial aggregation of airway hyper-responsiveness (AHR) to methacoline among randomly chosen families in a rural community in Anqing, China. METHODS: Airway responsiveness (AR) to methacoline and related risk factors were assessed in each subject. We first modeled the within family correlation in AR and demonstrated the familial aggregation of this trait. Furthermore, we examined the effect size (e.g., odds ratio, OR) of this correlation in a "subsequent offspring model." RESULTS: The correlation coefficient is significantly positive for parent-offspring and offspring offspring pairs, but not significant in father-mother pairs, suggesting a genetic component. The strength of the relationships is in the order of father-offspring < mother-offspring < offspring-offspring. The OR of a positive AHR test for subsequent offspring who had mothers and an eldest sibling with positive AHR is 4.12 (95% CI, 1.72-9.87), compared with subsequent offspring whose mother and eldest sibling were negative in the test. CONCLUSION: Our study supports a familial clustering of AHR in a Chinese population, which points to a role for genetic factors. PMID- 16257360 TI - The relationships of gender, cigarette smoking, and hypertension with the risk of aneurysmal subarachnoid hemorrhage: a case-control study in Nagoya, Japan. AB - PURPOSE: To examine the relationship of gender, cigarette smoking, and a history of hypertension to the risk of aneurysmal subarachnoid hemorrhage (SAH), using a case-control study. METHODS: Case subjects consisted of a consecutive series of 201 patients with spontaneous SAH with aneurysm(s) confirmed by angiography and/or CT scan. One hospital and one community control subject was matched to each case by gender and age (+/- 2 years). Multiple conditional logistic regression was used to calculate the odds ratio (OR) and 95% confidence interval (CI) adjusted for potential confounders. RESULTS: Current smoking and a history of hypertension were each significantly associated with an increased risk of subarachnoid hemorrhage for men and women combined. There was also a non significant trend towards synergism between these two factors with respect to an increased risk of subarachnoid hemorrhage for each gender separately and both combined. A significantly increased risk was observed for a history of hypertension (adjusted OR, 3.5; 95% CI, 1.2-14.7) among men, for current smoking alone (adjusted OR, 2.9; 95% CI, 1.1-7.7), and a history of hypertension alone (adjusted OR, 2.6; 95% CI, 1.4-5.1) among women. CONCLUSIONS: Trends towards gender differences and synergism emerged in the relationship of cigarette smoking and a history of hypertension of the risk of SAH provides useful information for targeting individuals/populations in programs for the primary prevention of SAH by gender. PMID- 16257361 TI - Rheumatoid arthritis among women in the Agricultural Health Study: risk associated with farming activities and exposures. AB - PURPOSE: Farming has been associated with increased risk of rheumatoid arthritis (RA) in some studies, but specific causes have not been identified. We studied risk factors for RA in the Agricultural Health Study, a cohort of over 57,000 licensed pesticide applicators and their spouses. METHODS: We used a nested case control design, limited to female participants. Physician-confirmed cases (n = 135) were matched to five controls each (n = 675) by birth date. We used logistic regression, adjusting for birth date and state to examine associations, as estimated by odds ratios (OR) and 95% confidence intervals (CI). RESULTS: Risk of RA was not associated with mixing or applying pesticides overall or with any pesticide class, nor did it vary by number of days or years of use. Certain pesticides were associated with small nonsignificantly increased risks, including lindane (OR = 1.8, 95% CI: 0.6-5.0). RA risk was associated with welding (OR = 2.1, 95% CI: 0.8-5.4), albeit imprecisely, but not with solvents or sunlight. CONCLUSIONS: We did not identify any strong risk factors for RA. Because of the severe disability associated with this relatively common disease, further investigation into causes is warranted both in the Agricultural Health Study and elsewhere. PMID- 16257363 TI - Dietary patterns and the risk of breast cancer. AB - PURPOSE: Ecologic, migrant, and secular trend studies suggest an association between the Western diet and the risk of breast cancer. We examined data from the Nurses Health Study II to evaluate the association between major dietary patterns and the risk of breast cancer among pre-menopausal women. METHODS: We derived "Western" and "prudent" dietary patterns from the responses of 90,638 premenopausal women, aged 26 to 46 years at baseline in 1991, who completed validated food frequency questionnaires in 1991 and 1995. These patterns were then evaluated for their associations with risk of breast cancer. RESULTS: During 8 years of follow-up, we documented 710 cases of invasive breast cancer. The multivariate RR (95% CI) comparing highest to lowest quintiles of cumulative average score were 0.90 (0.68-1.18, p-value, test for trend = 0.54) for the prudent dietary pattern and 0.97 (0.71-1.33, p-value, test for trend = 0.97) for the Western dietary pattern. CONCLUSIONS: Our findings suggest that there is no overall association between dietary patterns and risk of breast cancer. However, we found an inverse association between the prudent dietary pattern and breast cancer risk among ever smokers, but the test for interaction was not significant. We suggest that this finding merits further evaluation. PMID- 16257362 TI - Comparison of male and female breast cancer incidence trends, tumor characteristics, and survival. AB - PURPOSE: To compare male and female breast cancer and to determine the predictors of tumor characteristics and survival in both genders. METHODS: Male (n = 2923) and female breast cancer cases (n = 442,500) from the Surveillance, Epidemiology and End Results (SEER) registry were analyzed. Joinpoint regression was performed to detect changes in incidence trends from 1973 to 2001. Multiple logistic regression was used to regress each of four outcome variables (STAGE, LATERALITY, ESTROGEN, and PROGESTERONE RECEPTOR STATUS) on four demographic variables. Cox proportional hazards regression modeling was used to determine significant predictors of death of breast cancer after adjusting for demographic factors. RESULTS: Both men and women aged less than 50 years were at higher risk for advanced breast cancers. Males were at higher risk than females for advanced tumors among non-whites. The risk of breast cancer death among all cases was lower for each 10-year increase in age by 2%, higher for those who are unmarried than for those who are married by 12% and 13% higher for non-whites than for whites. CONCLUSIONS: Some important gender differences were detected with respect to factors associated with tumor characteristics, but gender was not a significant predictor of survival after adjusting for the other demographic variables. PMID- 16257364 TI - Does living in a religiously affiliated neighborhood lower mortality? AB - PURPOSE: To examine the effects of living in religiously affiliated and unaffiliated neighborhoods on mortality risks above that of individual risk factors, to determine if this effect behaves in a dose-response manner, and to examine the interaction between community wealth and religious affiliation. METHODS: Multilevel modeling of data from the Israel Longitudinal Mortality Study was used to assess mortality differentials based on neighborhood religious affiliation. Data were analyzed for 141,683 individuals aged 45 to 89 years and living in 882 statistical areas. Overall, 29,709 deaths were reported during the 9.5-year follow-up period. RESULTS: After accounting for individual demographic and socioeconomic (SES) characteristics as well as area-SES, men and women living in religiously affiliated neighborhoods had lower mortality rates than those living in unaffiliated areas (odds ratio(men) = 0.75; 95% CI, 0.67-0.84; odds ratio(women) = 0.86; 95% CI, 0.67-0.96). For men, this relationship behaved in a dose-response manner. Furthermore, the beneficial effects on mortality of living in a religiously affiliated area were consistent across age groups, middle-aged and elderly. Lastly, effect modification of area-SES on area-religion was observed for women only, whereby for women living in higher-SES areas, religiosity had no effect on mortality. CONCLUSIONS: The characteristics of one's immediate neighborhood, namely, community wealth and religious affiliation, have valuable health implications that should be included when assessing mortality risks. PMID- 16257365 TI - Overcoming challenges of using blood samples with gene expression microarrays to advance patient stratification in clinical trials. AB - The advances in blood protocols enable improved identification of biomarkers based on RNA expression profiles and might assist us in realizing the promise of pharmacogenomics. PMID- 16257366 TI - Another piece in the molecular puzzle of obesity. AB - Gene therapy or investigating the genetic component of obesity specifically related to SH2-B might be a more worthwhile approach to treat the obese. PMID- 16257367 TI - Drug discovery veers off target. AB - As drug pipelines stagnate, one potential savior comes in the form of a new scientific paradigm: systems biology. PMID- 16257368 TI - Frog skin hope for HIV prevention. AB - The discovery that the frog peptides can kill HIV virus even when it is hidden in dendritic cells suggests that they could be developed as mucosal preventives. PMID- 16257369 TI - Glyn Edwards talks about cancer drug development at Antisoma. Interview by Ulrike Knies-Bamforth and Dan Huke. AB - We're an exclusively oncology development-focused company; we've put together a portfolio of projects, which are rather different from each other, which is our way of diversifying the systemic risk out of the portfolio. PMID- 16257370 TI - Biomanufacturing: a high-growth industry for North Carolina. AB - The sustained growth of biotechnology in North Carolina has made the state a worldwide leader in biotechnology. PMID- 16257371 TI - Computing chemistry on the web. AB - The development of on-line software tools is changing the way we traditionally perform our analysis in drug design, but will chemoinformatics be forever behind bioinformatics in this development? PMID- 16257372 TI - Novel treatment options for infectious exacerbations. AB - Herein, we detail characteristics shared between two common therapeutic approaches, macrolide antibiotic therapy and systemic corticosteroids, in an attempt to propose an alternative treatment paradigm. PMID- 16257373 TI - Keynote review: phosphodiesterase-4 as a therapeutic target. AB - Cyclic AMP (cAMP) is a key second messenger in all cells. It is compartmentalized within cells and its levels are controlled, as a result of spatially discrete signaling cassettes controlling its generation, detection and degradation. Underpinning compartmentalized cAMP signaling are approximately 20 members of the phosphodiesterase-4 (PDE4) family. The selective inhibition of this family generates profound, functional effects and PDE4 inhibitors are currently under development to provide potential, novel therapeutics for the treatment of inflammatory diseases, such as asthma, chronic obstructive pulmonary disease and psoriasis, as well as treating depression and serving as cognitive enhancers. Here, we delineate the range of PDE4 isoforms, their role in signaling, their structural biology and related preclinical and clinical pharmacology. PMID- 16257374 TI - Quantitative risk modelling for new pharmaceutical compounds. AB - The process of discovering and developing new drugs is long, costly and risk laden. Faced with a wealth of newly discovered compounds, industrial scientists need to target resources carefully to discern the key attributes of a drug candidate and to make informed decisions. Here, we describe a quantitative approach to modelling the risk associated with drug development as a tool for scenario analysis concerning the probability of success of a compound as a potential pharmaceutical agent. We bring together the three strands of manufacture, clinical effectiveness and financial returns. This approach involves the application of a Bayesian Network. A simulation model is demonstrated with an implementation in MS Excel using the modelling engine Crystal Ball. PMID- 16257375 TI - The rational design of vaccines. AB - This review provides an insight into the various opportunities for vaccine intervention, analysis of strategies for vaccine development, vaccine ability to modulate immune responses and resultant rational vaccine design. In addition, wider aspects are considered, such as biotechnological advances, advances in immunological understanding and host-pathogen interactions. The key question addressed here is, with all our research and understanding, have we reached a new echelon in vaccine development, that of rational design? PMID- 16257377 TI - Measurements of binding thermodynamics in drug discovery. AB - Thermodynamics governs the process of biomolecular recognition. The steps of characterizing, understanding and exploiting binding thermodynamics have the potential to contribute to an improved rational drug design process that is more robust and reliable. It is only relatively recently that instrumentation capable of direct and full thermodynamic characterization has been improved, giving impetus to the application of thermodynamic measurements in drug discovery. This review highlights current instruments and methods that can be employed to measure binding thermodynamics and their use in studies of biomolecular recognition and drug discovery. PMID- 16257376 TI - Drug design using the example of the complement system inhibitors' development. AB - Undesired activation of the complement system, a part of the immune system, is a major pathogenic factor contributing to various diseases, such as ischemia reperfusion injury, sepsis, asthma, allergic reactions, rheumatoid arthritis, Alzheimer's disease, myasthenia, multiple sclerosis and others. The history of the development of complement system inhibitors, preventing its destructive action on the body, represents the evolution of the main methods of drug design. This review illustrates the main approaches of drug design, ranging from screening and modification of natural products to structure-based ligand design, on the basis of complement inhibitors' creation. The current status of the field of complement inhibitors is also discussed. PMID- 16257378 TI - Metabolomics: from pattern recognition to biological interpretation. AB - Metabolomics is a technology that aims to identify and quantify the metabolome -- the dynamic set of all small molecules present in an organism or a biological sample. In this sense, the technique is distinct from metabolic profiling, which looks for target compounds and their biochemical transformation. The combination of both approaches is an emerging technique for the characterization of biological samples and for drug treatment. Metabolomics has proven to be very rapid and superior to any other post-genomics technology for pattern-recognition analyses of biological samples. Changing steady state concentrations and fluctuations of metabolites that occur within milliseconds are a result of biochemical processes such as signalling cascades: metabolomic techniques are instrumental in measuring these changes rapidly and sensitively. Metabolite data can be complemented by protein, transcript and external (environmental) data, thereby leading to the identification of multiple physiological biomarkers embedded in correlative molecular networks that are not approachable with targeted studies. PMID- 16257379 TI - Exploring the sounds of silence: RNAi-mediated gene silencing for target identification and validation. AB - Drug development begins with the identification and early preclinical validation of novel biological targets, a process often called 'target identification and validation'. This process usually uses various approaches, such as observations from literature and findings from animal or clinical studies, together with cutting edge molecular techniques that include analyses of gene and protein expression, interaction and function. The publication of the human genome has increased research in gene and protein expression analysis that, in combination with RNA interference technology, promises the evaluation of novel functions for known genes, as well as hitherto unknown or unstudied genes with functions relevant to disease. PMID- 16257380 TI - The impact of Life Science Identifier on informatics data. AB - Since the Life Science Identifier (LSID) data identification and access standard made its official debut in late 2004, several organizations have begun to use LSIDs to simplify the methods used to uniquely name, reference and retrieve distributed data objects and concepts. In this review, the authors build on introductory work that describes the LSID standard by documenting how five early adopters have incorporated the standard into their technology infrastructure and by outlining several common misconceptions and difficulties related to LSID use, including the impact of the byte identity requirement for LSID-identified objects and the opacity recommendation for use of the LSID syntax. The review describes several shortcomings of the LSID standard, such as the lack of a specific metadata standard, along with solutions that could be addressed in future revisions of the specification. PMID- 16257382 TI - Single-shot, multicycle suicide gene therapy by replication-competent retrovirus vectors achieves long-term survival benefit in experimental glioma. AB - Achieving therapeutically efficacious levels of gene transfer in tumors has been a major obstacle for cancer gene therapy using replication-defective virus vectors. Recently, replicating viruses have emerged as attractive tools for cancer therapy, but generally achieve only transitory tumor regression. In contrast to other replicating virus systems, transduction by replication competent retrovirus (RCR) vectors is efficient, tumor-selective, and persistent. Correlating with its efficient replicative spread, RCR vector expressing the yeast cytosine deaminase suicide gene exhibited remarkably enhanced cytotoxicity in vitro after administration of the prodrug 5-fluorocytosine. In vivo, RCR vectors replicated throughout preestablished primary gliomas without spread to adjacent normal brain, resulting in profound tumor inhibition after a single injection of virus and single cycle of prodrug administration. Furthermore, stable integration of the replicating vector resulted in persistent infection that achieved complete transduction of ectopic glioma foci that had migrated away from the primary tumor site. Thus, efficient and stable integration of suicide genes represents a unique property of the RCR vector that achieved multiple cycles of synchronous cell killing upon repeated prodrug administration, resulting in chronic suppression of tumor growth and prolonged survival. PMID- 16257383 TI - In vivo depletion of CD4+CD25+ regulatory T cells enhances the antigen-specific primary and memory CTL response elicited by mature mRNA-electroporated dendritic cells. AB - We previously described mRNA electroporation as an efficient gene delivery method to introduce tumor-antigens (Ag) into murine immature dendritic cells (DC). Here, we further optimize the protocol and evaluate the capacity of mRNA-electroporated DC as a vaccine for immunotherapy. First, the early DC maturation kinetics and the effect of different lipopolysaccharide incubation periods on the phenotypic maturation profile of DC are determined. Next, we show that either immature or mature DC are equally well electroporated and express and present the transgene at a comparable level after electroporation. We point out that the mRNA electroporation results in a negative effect on the interleukin (IL)-12p70, IL-6, and tumor necrosis factor-alpha secretion after maturation. Nevertheless, mRNA electroporated DC induce an effective cytotoxic T lymphocyte (CTL) response in vivo. Mature electroporated DC are significantly more potent in eliciting an Ag specific CD8+ CTL response compared to their immature electroporated counterparts. In addition, a significant improvement in CTL response is obtained both in the primary and in the memory effector phases when CD4+CD25+ regulatory T cells (Treg) are depleted in vivo prior to immunization. These findings are further substantiated in tumor protection experiments and hold convincing evidence for the merit of Treg cell depletion prior to immunization with mRNA electroporated DC. PMID- 16257384 TI - A Bayesian Neural Network approach to estimating the Energy Equivalent Speed. AB - To reduce the number and the gravity of accidents, it is necessary to analyse and reconstruct them. Accident modelling requires the modelling of the impact which in turn requires the estimation of the deformation energy. There are several tools available to evaluate the deformation energy absorbed by a vehicle during an impact. However, there is a growing demand for more precise and more powerful tools. In this work, we express the deformation energy absorbed by a vehicle during a crash as a function of the Energy Equivalent Speed (EES). The latter is a difficult parameter to estimate because the structural response of the vehicle during an impact depends on parameters concerning the vehicle, but also parameters concerning the impact. The objective of our work is to design a model to estimate the EES by using an original approach combining Bayesian and Neural Network approaches. Both of these tools are complementary and offer significant advantages, such as the guarantee of finding the optimal model and the implementation of error bars on the computed output. In this paper, we present the procedure for implementing this Bayesian Neural Network approach and the results obtained for the modelling of the EES: our model is able to estimate the EES of the car with a mean error of 1.34 m s(-1). Furthermore, we built a sensitivity analysis to study the relevance of model's inputs. PMID- 16257385 TI - Optical storage and surface-relief gratings in azobenzene-containing nanostructured films. AB - This paper brings an overview of photoisomerization-derived properties in azobenzene-containing nanostructured films produced with the Langmuir-Blodgett (LB) and layer-by-layer (LbL) methods. Emphasis was placed on the optical storage and formation of surface-relief gratings (SRGs), where the distinctive properties of the nanostructured films were highlighted. For optical storage, in particular, a discussion was made of the higher birefringence induced in LB films from azopolymers due to their organized nature, and of the strong effects from ionic interactions on the photoisomerization of azochromophores in LbL films. The molecular-level control of film properties in the LbL films is described as being responsible for the varied behavior observed in the photoinscription of SRGs, which may involve considerable level of photodegradation depending on the film fabrication conditions and materials used. PMID- 16257386 TI - A novel culture system for porcine odontogenic epithelial cells using a feeder layer. AB - The growth of cells in vitro can provide useful models for investigating their behaviour and improving our understanding of their function in vivo. Although the developmental regulation of enamel matrix formation has been comprehensively analysed, the detailed cellular characteristics of ameloblasts remain unclear because of the lack of a system of long-term in vitro culture. Therefore, the establishment of odontogenic epithelial cell lines has taken on a new significance. Here, we report on a novel porcine odontogenic epithelial cell culture system, which has permitted serial culture of these cells. Epithelial cells were harvested from third molar tooth buds in the fresh mandibles of 6 month-old pigs, and seeded on dishes in D-MEM containing 10% FBS. Before the cells reached confluence, the medium was changed to LHC-9 to select the epithelial cells. When trypsinized epithelial cells were plated together with 3T3 J2 cells as a feeder layer, the epithelial cells grew from single cells into colonies. The colonies then expanded and became confluent, and could be sub cultured for up to 20 passages. The long-term culture cells expressed mRNA for amelogenin and ameloblastin, as well as enamelysin (MMP-20), which is a tissue specific gene product unique to ameloblasts. These results show that the system is capable of sustaining the multiplication of odontogenic epithelial cells with the characteristics of ameloblasts. PMID- 16257387 TI - Psychometric properties of the social phobia inventory: further evaluation. AB - This study investigated the psychometric properties of the Social Phobia Inventory [SPIN; Connor, K. M., Davidson, J. R. T., Churchill, L. E., Sherwood, A., Foa, E., Wesler, R.H., 2000. Psychometric properties of the Social Phobia Inventory (SPIN). British Journal of Psychiatry, 176, 379-386], a measure of severity in social phobia (social anxiety disorder). Participants included 132 participants with social phobia, 57 participants with panic disorder and agoraphobia (PDA), and 62 participants with obsessive-compulsive disorder (OCD). Confirming findings from an initial validation study, the SPIN was found to have excellent internal consistency and good test-retest reliability. It also distinguished well between those with social phobia and those with either PDA or OCD. Good convergent and discriminant validity were established by examining correlations with other conceptually related and unrelated scales. Finally, the SPIN was sensitive to changes in social phobia severity following cognitive behavioral group treatment. In conclusion, the SPIN is both reliable and valid for the measurement of social phobia severity and outcome following psychological treatment. PMID- 16257389 TI - Reduced expression of kinase-associated phosphatase in cortical dendrites of MAP2 deficient mice. AB - We previously demonstrated that cAMP-dependent protein kinase was reduced in the dendrites of MAP2-deficient mice. In this study, we compared the expression of various protein phosphatases (PPs) between wild-type and map2(-/-) dendrites. Kinase-associated phosphatase (KAP) was the only PP which showed difference between the two phenotypes: (1) the expression of KAP was reduced in map2(-/-) cortical dendrites, and (2) the amount of KAP bound to microtubules was reduced in map2(-/-) brains. We also demonstrated in cultured neuroblastoma cells that KAP is not only expressed in dividing cells, but also in the neurites of differentiated cells. Our findings propose that KAP, which has been reported to function in cell-cycle control, has an as yet uncovered role in regulating dendritic functions. We also propose MAP2-deficient mice as an ideal system for identifying protein phosphatases essential for dendritic functions. PMID- 16257388 TI - The impact of manipulating personal standards on eating attitudes and behaviour. AB - The relationship between perfectionism and eating disorders is well established and is of theoretical interest. This study used an experimental design to test the hypothesis that manipulating personal standards, a central feature of perfectionism, would influence eating attitudes and behaviour. Forty-one healthy women were randomly assigned either to a high personal standards condition (n = 18) or to a low personal standards condition for 24 h (n = 23). Measures of personal standards, perfectionism, and eating attitudes and behaviour were taken before and after the experimental manipulation. The manipulation was successful. After the manipulation, participants in the high personal standards condition ate fewer high calorie foods, made more attempts to restrict the overall amount of food eaten, and had significantly more regret after eating than those in the low personal standards condition. Other variables remained unchanged. It is concluded that experimental analyses can be of value in elucidating causal connections between perfectionism and eating attitudes and behaviour. PMID- 16257390 TI - Isothiazole dioxide derivative 6n inhibits vascular smooth muscle cell proliferation and protein farnesylation. AB - Isothiazole dioxides have been shown to inhibit Trypanosoma brucei protein farnesyltransferase (PFTase) in isolated enzyme, but elicited only a minor effect on mammalian PFTase. In the present study we have evaluated the effect of 3 diethylamino-4-(4-methoxyphenyl)-isothiazole 1,1-dioxides with different substituents at C5, on rat PFTase and protein geranylgeranyltransferase-I (PGGTase-I) with the final aims to improve the potency against mammalian PFTase and to identify new compounds with antiproliferative properties. For these purposes, in vitro and cell culture models have been utilized. The results showed that isothiazole dioxides with C4-C5 double bond and sulfaryl substituted at the C5 position but none of the dihydro-derivatives, were able to inhibit in vitro PFTase in a concentration dependent manner (IC50 ranging from 8.56 to 1015 microM). Among those, compound 6n (C5; methyl-S) displayed 500-fold higher inhibitory potency on PFTase than PGGTase-I. Compound 6n was shown to affect rat smooth muscle cell (SMC) proliferation at concentrations similar (IC50 = 61.4 microM) to those required to inhibit [3H]-farnesol incorporation into cellular proteins (-44.1% at 100 microM). Finally, compound 6n interferes with rat SMC proliferation by blocking the progression of G0/G1 phase without inducing apoptosis, as assessed by [3H]-thymidine incorporation assay and flow cytometry analysis. Taken together, we described a new PFTase inhibitor containing the isothiazole dioxide moiety that affects mammalian protein farnesylation and SMC proliferation by inhibiting G0/G1 phase of the cell cycle. PMID- 16257391 TI - Kinetics of tienilic acid bioactivation and functional generation of drug-protein adducts in intact rat hepatocytes. AB - Drug-induced autoimmune hepatitis is among the most severe hepatic idiosyncratic adverse drug reactions. Considered multifactorial, the disease combines immunological and metabolic aspects, the latter being to date much better known. As for many other model drugs, studies on tienilic acid (TA)-induced hepatitis have evidenced the existence of bioactivation during the hepatic oxidation of the drug, allowing the identification of the neoantigen of anti-LKM2 autoantibodies and the pathway responsible for its formation. However, most of these results are based on the use of microsomal fractions whose relevance to the liver in vivo still needs to be established. In the more complex intact cell environment, several endogenous processes may play a significant role on triggering the reaction and should therefore be considered. In this work we have characterised the kinetics of TA biotransformation in metabolically competent hepatocytes, the influence of TA bioactivation on physiological GSH levels, and the qualitative and quantitative profile of drug-protein conjugates generated in situ, as a function of exposure time. Results confirm that intact hepatocytes reproduce in vitro the metabolic sequence that leads to the functional generation of drug protein adducts, in conditions that simulate clinical human exposure to TA. Metabolically competent cultured hepatocytes appear as a very promising approach to investigate the early preimmunological events of drug-induced autoimmune hepatitis, adequate to identify the conditions that may modulate the formation and specificity of drug-protein adducts in vivo, to study the hepatic disposition of the TA-protein targets, and to define the specific role of the hepatocyte in the origin of this adverse reaction. PMID- 16257392 TI - l-DOPA administration enhances 6-hydroxydopamine generation. AB - The therapeutic success of L-3,4-dihydroxyphenylalanine (L-DOPA) treatment in Parkinson's disease (PD) patients remains controversial as many patients become tolerant requiring higher dosage regimens. However, the increase in dosage regimens results in the patients experiencing intolerable side effects. This study sought to investigate whether dopamine (DA) can chemically react with iron to form the potent neurotoxin 6-hydroxydopamine (6-OHDA). Furthermore, rats were treated with L-DOPA for a period of 7 and 28 days to determine whether L-DOPA treatment results in 6-OHDA formation in rat striatum. In addition, this study also investigates the complex interactions of L-DOPA with iron by performing in vitro and in vivo lipid peroxidation studies and the detection of endogenous 6 OHDA in iron-infused rats. In each study, melatonin was used to determine whether it could quench any free radical effects that may occur. The results of the present study show that DA chemically reacts with iron to form 6-OHDA. Moreover, L-DOPA treatment results in endogenous 6-OHDA formation in rat brain as well as enhances iron-induced lipid peroxidation both in vitro and in vivo in the rat striatum. The L-DOPA-induced increase in lipid peroxidation, in iron-infused rats, corresponds with an increase in levels of 6-OHDA in the rat striatum. The use of melatonin significantly decreases the L-DOPA-stimulated 6-OHDA formation in the rat striatum. The present study provides novel information on L-DOPA induced neurotoxicity and suggests the concomitant use of an antioxidant with L DOPA in order to enhance the life span of L-DOPA therapy. PMID- 16257393 TI - Effects of MCH and a MCH1-receptor antagonist on (palatable) food and water intake. AB - Melanin concentrating hormone (MCH) is a regulator of ingestive behavior, but several issues regarding its effects on specific components of ingestive behavior remain to be elucidated. Therefore, we injected, in the 3rd ventricle of male Wistar rats, saline, MCH (5 microg), MCH (5 mug) together with a MCH1-R antagonist (A, 10 microg) and the antagonist alone (A, 10 microg). Our results show that (1) central administration of MCH stimulates food intake (lab chow and medium high fat diet) and this can be blocked by a MCH1-R antagonist; (2) the MCH induced increase in food intake is mediated through increased meal number, meal duration and meal size; (3) the MCH1-R antagonist is able to significantly reduce the intake of a highly palatable food (condensed sweet milk) and is more effective in blocking MCH-induced food intake when rats are fed a palatable medium high fat food; and (4) MCH stimulated water intake independently from and disproportionately to food intake. In conclusion, our results point to an involvement of endogenous MCH in the enhanced intake of palatable food. Furthermore, they confirm that MCH stimulates not only food intake but also water intake. PMID- 16257394 TI - Marked differences in the efficacy of post-insult gene therapy with catalase versus glutathione peroxidase. AB - It is now recognized that the generation of reactive oxygen species (ROS) following necrotic neurological insults plays a central role in the subsequent neuron death. A key step in ROS detoxification is the conversion of hydrogen peroxide to water and oxygen by either catalase (CAT) or glutathione peroxidase (GPX). We have previously shown that overexpression of CAT or GPX protects cultured neurons against subsequent excitotoxic insults. Because of the unpredictability of most acute neurological insults, gene therapy will most often need to be carried out after rather than in anticipation of an insult. Thus, we have tested whether herpes virus amplicon vectors expressing CAT or GPX still protect cultured hippocampal neurons from oxygen/glucose deprivation if introduced following an insult. CAT-expressing vectors were protective even when introduced 8 h post-insult. In contrast, there was no post-insult time window in which GPX overexpression protected. While CAT requires no cofactor, GPX action requires glutathione as a cofactor. Thus, we speculated that the post-insult decline in glutathione compromises the protective potential of GPX. Supporting this, reversing the post-insult glutathione decline with glutathione supplementation was neuroprotective. PMID- 16257395 TI - Alterations in the dendritic morphology of prefrontal pyramidal neurons in adult rats after blockade of NMDA receptors in the postnatal period. AB - The present study assessed whether the blockade of NMDA receptors in the postnatal period, used to model the symptoms of schizophrenia altered morphology of pyramidal neurons in the medial prefrontal cortex of rats. CGP 40116, an antagonist of NMDA receptors, was given postnatally (days 1-21 after birth). The analysis of the morphology of pyramidal neurons visualized by the Golgi-Cox technique revealed that the exposure to an antagonist of NMDA receptors in the postnatal period diminished the length of basilar dendrites, while that of apical dendrites remained unchanged. The number of dendritic branches and the spine density remained unchanged. It is concluded that the blockade of NMDA receptors in the postnatal period only partially models morphological changes in pyramidal neurons of the medial prefrontal cortex, which are observed in some cases of schizophrenia. PMID- 16257396 TI - Interaction between organophosphate compounds and cholinergic functions during development. AB - Organophosphate (OP) compounds exert inhibition on cholinesterase (ChE) activity by irreversibly binding to the catalytic site of the enzymes. For this reason, they are employed as insecticides for agricultural, gardening and indoor pest control. The biological function of the ChE enzymes is well known and has been studied since the beginning of the XXth century; in particular, acetylcholinesterase (AChE, E.C. 3.1.1.7) is an enzyme playing a key role in the modulation of neuromuscular impulse transmission. However, in the past decades, there has been increasing interest concerning its role in regulating non neuromuscular cell-to-cell interactions mediated by electrical events, such as intracellular ion concentration changes, as the ones occurring during gamete interaction and embryonic development. An understanding of the mechanisms of the cholinergic regulation of these events can help us foresee the possible impact on environmental and human health, including gamete efficiency and possible teratogenic effects on different models, and help elucidate the extent to which OP exposure may affect human health. The chosen organophosphates were the ones mainly used in Europe: diazinon, chlorpyriphos, malathion, and phentoate, all of them belonging to the thionophosphate chemical class. This research has focused on the comparison between the effects of exposure on the developing embryos at different stages, identifying biomarkers and determining potential risk factors for sensitive subpopulations. The effects of OP oxonisation were not taken into account at this level, because embryonic responses were directly correlated to the changes of AChE activity, as determined by histochemical localisation and biochemical measurements. The identified biomarkers of effect for in vitro experiments were: cell proliferation/apoptosis as well as cell differentiation. For in vivo experiments, the endpoints were: developmental speed, size and shape of pre-gastrula embryos; developmental anomalies on neural tube, head, eye, heart. In all these events, we had evidence that the effects are mediated by ion channel activation, through the activation/inactivation of acetylcholine receptors (AChRs). PMID- 16257397 TI - The C-terminal peptides of acetylcholinesterase: cellular trafficking, oligomerization and functional anchoring. AB - In vertebrates, the catalytic domain of acetylcholinesterase (AChE) may be associated with several C-terminal peptides generated by alternative splicing in the 3' region of transcripts. The "readthrough" (R) variant results from a lack of splicing after the last exon encoding the catalytic domain. Such a variant has been observed in Torpedo and in mammals; its C-terminal r peptide, also called "AChE Related Peptide" (ARP), is poorly conserved between rodents and humans. In rodents, it is significantly expressed in embryonic tissues and at a very low level in the brain of adult mice; it may be increased under various stress conditions, but remains very low. The "hydrophobic" (H) variant generates glycolipid (GPI)-anchored dimers, which are expressed in muscles of Torpedo, and in blood cells of mammals; H variants exist in Torpedo and in mammals, but apparently not in other vertebrate classes, suggesting that they were lost during evolution of early vertebrates and re-appeared independently in mammals. The "tailed" (T) variant exists in all vertebrate cholinesterases and their C terminal t peptides are strongly conserved; in mammals, AChE(T) subunits represent the major type of acetylcholinesterase in cholinergic tissues. They produce a wide variety of oligomeric forms, ranging from monomers to heteromeric assemblies containing the anchoring proteins ColQ (collagen-tailed forms) and PRiMA (membrane-bound tetramers), which constitute the major functional enzyme species in mammalian muscles and brain, respectively. The oligomerization of AChE(T) subunits depends largely on the properties of their C-terminal t peptide. These peptides contain seven conserved aromatic residues, including three tryptophans, and are organized in an amphiphilic alpha helix in which these residues form a hydrophobic cluster. The presence of a cysteine is required for dimerization, while aromatic residues are necessary for tetramerization. In the collagen-tailed molecules, four t peptides form a coiled coil around a proline rich motif (PRAD) located in the N-terminal region of ColQ. The t peptide also strongly influences the folding and cellular trafficking of AChE(T) subunits: the presence of hydrophobic residues induces partial misfolding leading to inactive protein, while aromatic residues, organized or not in an amphiphilic helix, induce intracellular degradation through the "Endoplasmic Reticulum Associated Degradation" (ERAD) pathway, rather than secretion. It has been proposed that the r and t C-terminal peptides, or fragments of these peptides, may exert independent, non cholinergic biological functions: this interesting possibility still needs to be documented, especially in view of their various degrees of evolutionary conservation. PMID- 16257399 TI - Review: Collagen markers in early arthritic diseases. AB - In arthritic diseases e.g. osteoarthritis (OA) and rheumatoid arthritis (RA), the stability of the collagen type II (CII) fibers, a major component of articular cartilage, is compromised with extensive proteolytic breakdown leading to cartilage erosion and joint deterioration. A clinical need for molecular markers that give instantaneous measure of rate of joint deterioration has developed, as other measurements e.g. arthroscopy, and joint space narrowing are insensitive to small changes in disease status over short periods of time. Owing to its exclusive presence in cartilaginous tissues, markers of CII synthesis and degradation have been extensively studied. Assays that measure these markers in biological fluids e.g. synovial fluid (SF), serum, and urine have been developed and applied to detect early disease onset, monitor disease progression, and response to anti-arthritic drugs. CII synthesis markers include the procollagen type II C-propeptide (PIICP) and the procollagen type IIA N-propeptide (PIIANP). CII degradation markers include CII C-telopeptide (CII-X), CII neoepitope (TIINE), helix II, C2C, CNBr 9.7, Coll 2-1, and Coll 2-1 NO(2). Most of these markers differentiate between early stages of OA, RA and reference controls. The best correlations with structural changes occur when measurements are made in SF while serum measurement frequently did not correlate with structural changes. Although the selection of an optimal marker or a set of markers is still problematic, few markers are of considerable utility in early detection and monitoring of arthritic diseases. The current challenge is to improve the discriminatory power of these markers so they can be used to guide therapeutic decisions. PMID- 16257398 TI - Inhibition of acetylcholinesterase by the anticancer prodrug CPT-11. AB - CPT-11 (irinotecan, 7-ethyl-10-[4-(1-piperidino)-1 piperidino]carbonyloxycamptothecin) is an anticancer prodrug that has been approved for the treatment of colon cancer. It is a member of the camptothecin class of drugs and activation to the active metabolite SN-38, is mediated by carboxylesterases (CE). SN-38 is a potent topoisomerase I poison and is highly effective at killing human tumor cells, with IC50 values in the low nM range. However, upon high dose administration of CPT-11 to cancer patients, a cholinergic syndrome is observed, that can be rapidly ameliorated by atropine. This suggests a direct interaction of the drug or its metabolites with acetylcholinesterase (AChE). Kinetic studies indicated that CPT-11 was primarily responsible for AChE inhibition with the 4-piperidinopiperidine moiety, the major determinant in the loss of enzyme activity. Structural analogs of 4 piperidinopiperidine however, did not inhibit AChE, including a benzyl piperazine derivate of CPT-11. These results suggest that novel anticancer drugs could be synthesized that do not inhibit AChE, or alternatively, that novel AChE inhibitors could be designed based around the camptothecin scaffold. PMID- 16257400 TI - Validation of (1)H NMR spectroscopy as an analytical tool for methylamine metabolites in urine. AB - BACKGROUND: Methylamines have many metabolic roles and there is an increasing demand for their measurement. Glycine betaine is an important osmolyte, and a reservoir for methyl groups. Proline betaine and trigonelline are important dietary betaines. Trimethylamine, derived from gut flora, is normally converted to trimethylamine oxide but in 'fish odour syndrome' is excreted as TMA. These compounds are all suitable for quantification by (1)H NMR spectroscopy as they all have methyl protons. METHOD: Urine samples are acidified and (1)H NMR spectra are obtained using presaturation for water suppression. Peak integrals or heights are compared to an internal standard of acetonitrile. RESULTS: Inter- and intra assay CV's were <5% for TMAO and creatinine, and <10% for the other analytes. Responses were linear from 50 to 1000 microM for all metabolites, and recoveries were > or =97%. Limits of detection using NMR are slightly higher than alternative HPLC assays (15-25 microM). However, sensitivity is adequate for the detection of raised levels in urine, and sample analysis was complete in less than 5 min. CONCLUSION: (1)H NMR spectroscopy is a convenient, rapid and economical option for the determination of betaines and related compounds in urine in a single analysis. PMID- 16257401 TI - Lysosomal enzyme activities: new potential markers for Sjogren's syndrome. AB - OBJECTIVE: To evaluate the changes in lysosomal enzyme activities in leukocytes of patients with Sjogren's syndrome. METHODS: Leukocytes were obtained from 38 patients with Sjogren's syndrome and 36 healthy subjects. The activities of the following glycosidases were measured: alpha-glucosidase (AGU), beta-galactosidase (BGA), alpha-mannosidase (AMAN), beta-glucuronidase (GCU), beta-hexosaminidase (HEX), and the following proteases: cathepsin B (CATH B), dipeptidyl peptidase I (DPP I), cathepsin H (CATH H), dipeptidyl peptidase II (DPP II), tripeptidyl peptidase I (TPP I), and cathepsin D (CATH D) activity. RESULTS: Activity of the glycosidases beta-galactosidase, alpha-mannosidase, beta-glucuronidase and beta hexosaminidase, as well as of the peptidases cathepsin B, cathepsin D, dipeptidyl peptidase I, and tripeptidyl peptidase I, was elevated during the first 5 years of SS, and it increased further between 5 and 10 years after diagnosis. CONCLUSIONS: The elevated activities of the lysosomal enzymes in Sjogren's syndrome patients may play a role in tissue damage by accelerated breakdown of glycoproteins in lysosomes. PMID- 16257402 TI - Influence of the selective neuronal NO synthase inhibitor ARL 17477 on nitrergic neurotransmission in porcine stomach. AB - Selective neuronal NOS (nNOS) inhibitors have been developed for possible application in cerebral ischemia and neurodegenerative disorders. To investigate the degree of interference with peripheral nNOS, the influence of the selective nNOS inhibitor ARL 17477 was studied on electrically induced nitrergic relaxations in pig gastric fundus strips and on gastric fundic compliance in conscious pig. Circular muscle strips of porcine gastric fundus were electrically stimulated (10 s trains at 4 Hz, 0.1 ms and 40 V). ARL 17477 inhibited the electrically induced relaxations in a concentration-dependent way (3x10(-6) M-10( 4) M). The inhibitory effect of ARL 17477 developed more progressively than that of N(G)-nitro-L-arginine methyl ester (L-NAME; 3x10(-4) M). In conscious pigs, instrumented with a fundic cannula, L-NAME (20 mg/kg i.v.) significantly increased mean arterial blood pressure and decreased fundic compliance in the fasted state (71+/-13 ml/mm Hg versus 185+/-37 ml/mm Hg after saline; P<0.05). ARL 17477 (3 mg/kg, i.v.) did not influence blood pressure but influenced gastric fundic volume-pressure curves in a similar way as L-NAME. Plasma concentration analysis of ARL 17477 indicated a half-life of less than 30 min in pig. ARL 17477 thus inhibits the effect of nitrergic neurons in the pig gastric fundus in vitro, leading to inhibited gastric compliance in the conscious pig. The study indicates that selective nNOS inhibitors, applied for cerebral disorders, might also interfere with neuronal nitrergic regulation of gastrointestinal motility. PMID- 16257403 TI - p16INK4A expression in cervical premalignant and malignant lesions. AB - p16INK4a is a cyclin-dependent kinase (CDK) inhibitor which decelerates cell cycle by inactivating CDKs that phosphorylate pRb. Human Papillomavirus persistent infection plays an important role on cervical carcinogenesis, mainly by the action of two viral oncoproteins, E6 and E7, which interact with p53 and pRb, respectively. Increasing expression of E6 and E7 in dysplastic cervical cells might thus be reflected by increased expression of p16INK4a. Recent studies revealed that p16INK4a expression could be a marker for dysplastic and neoplastic cervical cells. The aim of this study was to analyze p16INK4a expression in cervical preneoplastic and neoplastic lesions and correlate with lesion grade. Expression of p16INK4a was analyzed by immunohistochemistry. A total of 6 low grade squamous intraepithelial lesion (LSIL), 21 high-grade squamous intraepithelial lesions (HSIL) and 27 cancer samples were studied. In HPV positive cervical samples (n=48), p16INK4a expression was observed in 1 of 3 LSIL, in 18 of 19 HSIL and in all 26 cancer cases. These results are in accordance with the hypothesis that functional inactivation of pRb by HPV-E7 protein induces p16INK4a expression in cervical lesions. In our study, a statistically significant association was observed between cervical lesion grade and p16INK4a expression (P<0.001). PMID- 16257404 TI - Glucocorticoids of bison bulls in relation to social status. AB - A primary response to stress is an increase in circulating adrenal glucocorticoids (GC) such as cortisol. Two hypotheses propose differential stress responses to agonistic and aggressive interactions in social groups. If subordinate animals are subjected to social and psychological stressors leading to chronic GC elevation, the 'stress of subordination' hypothesis predicts that GCs will be higher in subordinates than dominants. Alternatively, if dominant animals are subject to physiological stressors (e.g., fight at higher rates than subordinates) or hierarchies are unstable, the 'stress of domination' hypothesis predicts higher GCs in dominant individuals. Both models predict that GC levels will peak during the breeding season. We tested these predictions in bison bulls (Bison bison) using fecal steroid analysis to characterize GC concentration and behavioral observations to determine dominance rank, copulatory success, and tending status of bulls. Fecal samples were collected during 2003 from adult bison bulls during pre-rut (June), rut (July-August), and post-rut (September). Matched sample data indicated that mean GC levels (ng/g feces) of bulls strongly peaked during the 4-week rut, doubling from pre-rut to rut and then declining again during post-rut. High ranked dominant bulls maintained higher GC levels than lower ranked subordinate bulls. Dominance rank was positively correlated with copulatory success and age, and dominant bulls were more likely to tend (guard) cows as they approached estrus. There was a positive correlation between GC level and copulatory success, with prime-aged bulls (> or =7 years) obtaining the most copulations. GC levels were positively correlated with bull androgen levels determined in a previous study. These results support the 'stress of domination' hypothesis, indicating that dominant bison bulls pay a significant physiological price for high social status and the opportunity to mate. PMID- 16257405 TI - Estradiol and progesterone differentially regulate formalin-induced nociception in ovariectomized female rats. AB - Clinical and preclinical studies have found sex-specific differences in the discrimination and perception of inflammatory stimuli. The emerging picture suggests that the biological basis of these differences resides in the regulatory activity of gonadal hormones in the central nervous system. This study describes the effects of ovarian hormones in inflammatory pain processes. Ovariectomized rats received estradiol and/or progesterone, and the number of paw flinches was measured after 1, 2.5 or 5% formalin administration. Both estradiol and progesterone altered the number of flinches only after 1% formalin administration. Estradiol significantly reduced the overall number of flinches during Phase II of the formalin nociceptive response while progesterone attenuated Phase I of the response. After co-administration of estradiol and progesterone, progesterone reversed estradiol's analgesic effect in Phase II, however, estradiol did not reverse progesterone's analgesic activity in Phase I. To determine if estradiol effects are receptor-mediated, tamoxifen (selective estrogen receptor mediator, 15 mg/kg) or alpha-estradiol (an inactive isomer of estradiol, 20 microg) were utilized. Tamoxifen decreased the number of formalin induced flinches during Phase II while alpha-estradiol did not affect any formalin-induced responses. When co-administered with estradiol, tamoxifen failed to reverse estradiol's effect, suggesting both tamoxifen and estradiol activate similar intracellular mechanisms. Although Western blot analysis detected the presence of estradiol alpha and beta and progesterone B receptors in the spinal cord, hormone replacement treatments had no effects on the levels of these receptors. We postulate that the mechanisms by which estradiol and progesterone induce analgesia occur through the activation of their receptor at the spinal cord level. PMID- 16257406 TI - Is a fat stitch required when closing a hip hemiarthroplasty wound without a drain? AB - Studies have shown no benefit of a subcutaneous fat stitch when closing hip wounds, but all have been in the presence of a drain. Our aim was to determine whether, in the absence of a drain, suturing or not of the subcutaneous fat layer in hip hemiarthroplasty wounds had any significant effect on wound complication rate. We performed a prospective cohort study of 45 hip hemiarthroplasty patients who had a fat stitch and 40 who did not. No drains were used in either group. There were 44 patients in the fat stitch group and 35 in the no fat stitch group after six patients were excluded. The infection rate was 2% for the fat stitch group, and 20% for the no fat stitch group (p=0.02). There were no cases of deep dehiscence in the fat stitch group, but four cases (11%) in the no fat stitch group (p=0.035). The overall complication rate in the fat stitch group was 6.8% compared to 33% in the no fat stitch group (p=0.007). In the absence of a drain, we have found a significant increase in hip hemiarthroplasty wound complications when the subcutaneous fat is not sutured. PMID- 16257407 TI - Nurse Practitioner competency standards: findings from collaborative Australian and New Zealand research. AB - BACKGROUND: The title, Nurse Practitioner, is protected in most jurisdictions in Australia and New Zealand and the number of nurse practitioners is increasing in health services in both countries. Despite this expansion of the role, there is scant national or international research to inform development of nurse practitioner competency standards. OBJECTIVES: The aim of this study was to research nurse practitioner practice to inform development of generic standards that could be applied for the education, authorisation and practice of nurse practitioners in both countries. DESIGN: The research used a multi-methods approach to capture a range of data sources including research of policies and curricula, and interviews with clinicians. Data were collected from relevant sources in Australia and New Zealand. SETTINGS: The research was conducted in New Zealand and the five states and territories in Australia where, at the time of the research, the title of nurse practitioner was legally protected. PARTICIPANTS: The research was conducted with a purposeful sample of nurse practitioners from diverse clinical settings in both countries. Interviews and material data were collected from a range of sources and data were analysed within and across these data modalities. RESULTS: Findings included identification of three generic standards for nurse practitioner practice: namely, Dynamic Practice, Professional Efficacy and Clinical Leadership. Each of these standards has a number of practice competencies, each of these competencies with its own performance indicators. CONCLUSIONS: Generic standards for nurse practitioner practice will support a standardised approach and mutual recognition of nurse practitioner authorisation across the two countries. Additionally, these research outcomes can more generally inform education providers, authorising bodies and clinicians on the standards of practice for the nurse practitioner whilst also contributing to the current international debate on nurse practitioner standards and scope of practice. PMID- 16257408 TI - C-reactive protein levels and coronary artery disease incidence and mortality in apparently healthy men and women: the EPIC-Norfolk prospective population study 1993-2003. AB - INTRODUCTION: Measurement of C-reactive protein (CRP) levels has been proposed as a useful marker to improve the prediction of future coronary artery disease (CAD) risk, but this notion has been challenged recently. METHODS AND RESULTS: We performed a prospective case-control study among apparently healthy men and women. The odds ratio (OR) for future CAD incidence was 2.49 (95% CI=2.02-3.08, p for linearity <0.0001) unadjusted, and 1.66 (95% CI=1.31-2.12, p for linearity <0.0001), after adjustment for classical cardiovascular risk factors, for top versus bottom quartile of the CRP distribution. Notably, the risk factor adjusted predictive value was substantially stronger for fatal CAD (OR=2.92, 95% CI=1.83 4.67, p for linearity <0.0001) than for non-fatal CAD (OR=1.25, 95% CI=0.93-1.66, p for linearity=0.06). CRP levels were among the strongest predictors of CAD incidence and mortality. CRP levels remained a statistically significant predictor of future CAD, even after adjustment for the Framingham risk score. CONCLUSIONS: In this British cohort with risk factor levels representative of a contemporary Western population, CRP concentration was among the strongest predictors of CAD incidence and mortality. We suggest that current guidelines on CRP measurement in clinical practice should be based on contemporary and representative populations. PMID- 16257409 TI - Determination of 2,4,6-trichloroanisole and guaiacol in cork stoppers by pressurised fluid extraction and gas chromatography-mass spectrometry. AB - This paper describes the development of an analytical method consisting of pressurised fluid extraction (PFE) and gas chromatography-mass spectrometry (GC MS) using experimental designs to determine two volatile compounds in naturally tainted cork stoppers. The target analytes, 2,4,6-trichloroanisole (2,4,6-TCA) and guaiacol, are involved in the cork taint of wine. First, a Plackett-Burman experimental design was carried out in order to determine the significant experimental parameters affecting the PFE process, and then a central composite design was used to optimise these significant parameters. Once the method had been optimised, the influence of the number of extraction cycles was studied. The method was applied to determine the concentration of 2,4,6-TCA and guaiacol in three cork samples, and the results were compared with the ones obtained by multiple headspace-solid-phase microextraction (MHS-SPME) and by Soxhlet extraction. PMID- 16257410 TI - Design and fabrication of integrated solid-phase extraction-zone electrophoresis microchip. AB - Integrated solid-phase extraction-zone electrophoresis (SPE-ZE) device has been designed and fabricated on microchip. The structures were fabricated by using multiple layers of SU-8 polymer with a novel technique that enables easy alignment and high yield of the chips. SU-8 adhesive bonding has two major advantages: it enables bonding of high aspect ratio pillars and it results in fully SU-8 microchannels with uniform electrokinetic flow properties. The SPE-ZE device has a fluidic reservoir with 15:1 high aspect ratio pillars for bead filters that act as a SPE part in the chip structure. The separation unit is a 25 mm long electrophoresis channel starting from the outlet of SPE reservoir. Argon laser-induced fluorescence (LIF) detector was used to monitor simultaneously the SPE reservoir and the detection site at the end of the electrophoresis channel. Flow characteristics and electric field distributions were simulated with Femlab software. Fluorescein was used as the analyte for detecting the operational performance of the chip. Adsorption, bead rinsing, elution and detection were tested to verify functioning of the chip design. PMID- 16257411 TI - Effect of anoxia on diapause termination in eggs of the false melon beetle, Atrachya menetriesi. AB - The effect of anoxia on diapause development in the leaf beetle Atrachya menetriesi was investigated to elucidate the role of oxygen in regulation of egg diapause. While anoxia alone had no effect on diapause termination, it decreased diapause intensity before chilling. Such an effect reached a maximum level when anoxia lasted for about 10 days. Anoxia applied during the pre-diapause stage also reduced diapause intensity. On the other hand, anoxia terminated diapause when the diapause intensity had been lowered by sufficient duration of chilling (50 days at 7.5 degrees C). The effect of anoxia was temperature dependent; the larger effect was elicited when anoxia was combined with a higher temperature. A 50-day chilling caused more than 20% of eggs to terminate diapause upon transfer to warm conditions. However, when this chilling period was interrupted on the 20th day by a 5-day exposure to a high temperature of 20, 25 or 30 degrees C, the effect of the former chilling was cancelled partially or completely, suggesting that warming reversed diapause development. This reversing effect of a high temperature, however, was not manifested when the warming was combined with anoxia. The results suggest that anoxia inhibits diapause reversal and facilitates a certain process of diapause development. The sequence of exposure to anoxia and chilling is not important. PMID- 16257412 TI - Complexity of the cold acclimation response in Drosophila melanogaster. AB - Insects can increase their resistance to cold stress when they are exposed to non lethal conditions prior to the stress; these plastic responses are normally described only in terms of immediate effects on mortality. Here we examine in Drosophila melanogaster the short- and longer-term effects of different conditions on several measures of cold resistance, but particularly chill coma recovery. Short-term exposure to sublethal temperature (cold hardening) did not decrease chill coma recovery times even though it decreased mortality. Exposure to 12 degrees C for 2 days (acclimation) decreased chill coma recovery times for a range of stressful temperatures when flies were cultured at 25 degrees C, but did not usually affect recovery times when flies were cultured at 19 degrees C. In contrast, 2-day exposure to 12 degrees C decreased mortality regardless of rearing temperature. Rearing at 19 degrees C decreased mortality and chill coma recovery time relative to rearing at 25 degrees C. Acclimation increased the eclosion rate of eggs from stressed females, but did not affect development time or size of the offspring. These results indicate that plastic responses to cold in D. melanogaster are complex when resistance is scored in different ways, and that effects can extend across generations. PMID- 16257413 TI - Improvement of a FRET-based indicator for cAMP by linker design and stabilization of donor-acceptor interaction. AB - Forster resonance energy transfer (FRET) technology has been used to develop genetically encoded fluorescent indicators for a variety of intracellular molecular events. Often, however, the poor dynamic range of such reporters prevents detection of subtle but physiologically relevant signals. Here we present a strategy for improving FRET efficiency between donor and acceptor fluorophores in a green fluorescent protein (GFP)-based protein indicator for cAMP. Such indicator is based on protein kinase A (PKA) and was generated by fusion of CFP and YFP to the regulatory and catalytic subunits of PKA, respectively. Our approach to improve FRET efficiency was to perform molecular dynamic simulations and modelling studies of the linker peptide (L11) joining the CFP moiety and the regulatory subunit in order to define its structure and use this information to design an improved linker. We found that L11 contains the X-Y P-Y-D motif, which adopts a turn-like conformation that is stiffly conserved along the simulation time. Based on this finding, we designed a new linker, L22 in which the YPY motif was doubled in order to generate a stiffer peptide and reduce the mobility of the chromophore within the protein complex, thus favouring CFP/YFP dipole-dipole interaction and improving FRET efficiency. Molecular dynamic simulations of L22 showed, unexpectedly, that the conformational behaviour of L22 was very loose. Based on the analysis of the three principal conformational states visited by L22 during the simulation time, we modified its sequence in order to increase its rigidity. The resulting linker L20 displayed lower flexibility and higher helical content than L22. When inserted in the cAMP indicator, L20 yielded a probe showing almost doubled FRET efficiency and a substantially improved dynamic range. PMID- 16257414 TI - The energy landscape of modular repeat proteins: topology determines folding mechanism in the ankyrin family. AB - Proteins consisting of repeating amino acid motifs are abundant in all kingdoms of life, especially in higher eukaryotes. Repeat-containing proteins self organize into elongated non-globular structures. Do the same general underlying principles that dictate the folding of globular domains apply also to these extended topologies? Using a simplified structure-based model capturing a perfectly funneled energy landscape, we surveyed the predicted mechanism of folding for ankyrin repeat containing proteins. The ankyrin family is one of the most extensively studied classes of non-globular folds. The model based only on native contacts reproduces most of the experimental observations on the folding of these proteins, including a folding mechanism that is reminiscent of a nucleation propagation growth. The confluence of simulation and experimental results suggests that the folding of non-globular proteins is accurately described by a funneled energy landscape, in which topology plays a determinant role in the folding mechanism. PMID- 16257415 TI - Exploring the mechanical behavior of single intermediate filaments. AB - Intermediate filaments (IFs) are structural elements of eukaryotic cells with distinct mechanical properties. Tissue integrity is severely impaired, in particular in skin and muscle, when IFs are either absent or malfunctioning due to mutations. Our knowledge on the mechanical properties of IFs is mainly based on tensile testing of macroscopic fibers and on the rheology of IF networks. At the single filament level, the only piece of data available is a measure of the persistence length of vimentin IFs. Here, we have employed an atomic force microscopy (AFM) based protocol to directly probe the mechanical properties of single cytoplasmic IFs when adsorbed to a solid support in physiological buffer environment. Three IF types were studied in vitro: recombinant murine desmin, recombinant human keratin K5/K14 and neurofilaments isolated from rat brains, which are composed of the neurofilament triplet proteins NF-L, NF-M and NF-H. Depending on the experimental conditions, the AFM tip was used to laterally displace or to stretch single IFs on the support they had been adsorbed to. Upon applying force, IFs were stretched on average 2.6-fold. The maximum stretching that we encountered was 3.6-fold. A large reduction of the apparent filament diameter was observed concomitantly. The observed mechanical properties therefore suggest that IFs may indeed function as mechanical shock absorbers in vivo. PMID- 16257416 TI - An intricate RNA structure with two tRNA-derived motifs directs complex formation between yeast aspartyl-tRNA synthetase and its mRNA. AB - Accurate translation of genetic information necessitates the tuned expression of a large group of genes. Amongst them, controlled expression of the enzymes catalyzing the aminoacylation of tRNAs, the aminoacyl-tRNA synthetases, is essential to insure translational fidelity. In the yeast Saccharomyces cerevisiae, expression of aspartyl-tRNA synthetase (AspRS) is regulated in a process necessitating recognition of the 5' extremity of AspRS messenger RNA (mRNA(AspRS)) by its translation product and adaptation to the cellular tRNA(Asp) concentration. Here, we have established the folding of the approximately 300 nucleotides long 5' end of mRNA(AspRS) and identified the structural signals involved in the regulation process. We show that the regulatory region in mRNA(AspRS) folds in two independent and symmetrically structured domains spaced by two single-stranded connectors. Domain I displays a tRNA(Asp) anticodon-like stem-loop structure with mimics of the aspartate identity determinants, that is restricted in domain II to a short double-stranded helix. The overall mRNA structure, based on enzymatic and chemical probing, supports a three-dimensional model where each monomer of yeast AspRS binds one individual domain and recognizes the mRNA structure as it recognizes its cognate tRNA(Asp). Sequence comparison of yeast genomes shows that the features within the mRNA recognized by AspRS are conserved in different Saccharomyces species. In the recognition process, the N-terminal extension of each AspRS subunit plays a crucial role in anchoring the tRNA-like motifs of the mRNA on the synthetase. PMID- 16257417 TI - The good, the bad and the discriminator--errors in direct and indirect reciprocity. AB - This paper presents, in a series of simple diagrams, concise results about the replicator dynamics of direct and indirect reciprocity. We consider repeated interactions between donors and recipients, and analyse the relationship between three basic strategies for the donor: unconditional cooperation, all-out defection, and conditional cooperation. In other words, we investigate the competition of discriminating and indiscriminating altruists with defectors. Discriminators and defectors form a bistable community, and hence a population of discriminators cannot be invaded by defectors. But unconditional altruists can invade a discriminating population and 'soften it up' for a subsequent invasion by defectors. The resulting dynamics exhibits various forms of rock-paper scissors cycles and depends in subtle ways on noise, in the form of errors in implementation. The probability for another round (in the case of direct reciprocity), and information about the co-player (in the case of indirect reciprocity), add further elements to the ecology of reciprocation. PMID- 16257418 TI - Tissue-specific reactivation of gene expression at an imprinted locus. AB - Genomic imprinting is the phenomenon where the expression pattern of an allele at a locus differs depending on the allele's parent of origin. In most cases, one of the two alleles is transcriptionally silent. Recent empirical work has shown some genes to be imprinted in a tissue-specific manner, where the silenced allele becomes reactivated in particular cell lineages during development. Here I describe an evolutionary model of tissue-specific transcriptional reactivation. The model describes the relationships among various inclusive fitness functions and phenotypic effects necessary for natural selection to favor the epigenetic reprogramming required for this sort of reactivation, and makes predictions regarding the nature and magnitude of phenotypic and fitness consequences of mutations in particular somatic tissues. In particular, if an imprinted gene is reactivated in one of two tissues that interact in producing a particular phenotype, expression of the gene in those two tissues is expected to have opposite phenotypic effects. The model predicts that in some cases, mutations affecting the silenced allele at an imprinted locus may be phenotypically more severe than those affecting the expressed allele. These predictions are contrasted with those of an alternative explanation for reactivation: protection against deleterious recessive somatic mutations. The inclusive-fitness model of reactivation indicates that the intragenomic conflicts present in the parental germ lines and developing embryo persist though adult life, and can have complex effects on phenotypes and patterns of gene expression in somatic tissues. PMID- 16257419 TI - Effects of excess salt and fat intake on myocardial function and infarct size in rat. AB - Important risk factors for cardiovascular disease include excess dietary intake of saturated fat and (or) salt. This study tested the hypothesis that excess intakes of saturated fat (e.g., beef tallow) and salt cause greater myocardial cell death following ischemia-reperfusion injury than each risk factor alone. Male rats were divided into four groups: basal fat diet (4.5% as calories; control), high fat diet (40% as calories; FAT), basal fat diet and high salt (1% NaCl solution; SALT) and high fat diet and high salt (FATSALT). The gain in body weight was significantly higher for FAT and FATSALT groups than those of either the control or the SALT group. Five weeks of exposure to the dietary regimens did not significantly affect the coronary flow rate and except for the salt-fed group, had no effect on the rate-pressure-product of the isolated heart perfused in Langendorff mode. Although infarct size was not affected by the high fat diet, it was reduced by the high salt regimen relative to the high fat diet or the control groups. When rats were fed the FAT and SALT combination, the effect of salt feeding on infarct size was not observed. In addition, the FATSALT group displayed a more marked deterioration in contractile function following ischemia reperfusion injury than the other groups. In conclusion, short-term intake of a high fat diet, which significantly increases body weight, does not worsen ischemia-reperfusion injury although the treatment prevents the reduction of infarct size associated with high salt feeding. PMID- 16257421 TI - Diazepam effects on Ehrlich tumor growth and macrophage activity in mice. AB - Besides the central gabaergic receptors described for benzodiazepines, peripheral type binding sites (PBR) were also identified for these molecules in endocrine steroidogenic tissues, immune organs and cells, such as macrophages and lymphocytes. PBR activation was reported to decrease innate immunity and host defense. The present experiment was designed to analyze the effects of diazepam on Ehrlich tumor growth, and on macrophage activity of Ehrlich tumor bearing mice. Results showed that diazepam (3.0 mg/kg/day, for 7 days) increased the number of Ehrlich tumor cells and the volume of tumor-induced ascitic fluid. These effects were not observed after smaller doses of diazepam, suggesting a dose-dependant effect. Furthermore, our results show that 3.0 mg/kg of diazepam, administered daily, for 2 days, decreased (1) the number of peritoneal leukocytes retrieved after injection of the Ehrlich tumor, (2) the percents of macrophage spreading, and (3) the levels of macrophage NO production. Diazepam (3.0 mg/kg/day for 2 days) had no effect on macrophage phagocytosis or on H2O2 production. The present data is discussed based on a direct and/or indirect action of diazepam. Particularly, our findings might be due to a direct effect of diazepam on PBRs present on macrophages and tumor cells, or could still be mediated by PBR stimulation within the hypothalamus-pituitary-adrenal (HPA) axis. PMID- 16257420 TI - The dynamic relationship between mu and kappa opioid receptors in body temperature regulation. AB - Previous studies demonstrated that intracerebroventricular (icv) injection of a kappa opioid receptor agonist decreased, and a mu agonist increased, body temperature (Tb) in rats. A dose-response study with the selective kappa antagonist nor-binaltorphimine (nor-BNI) showed that a low dose (1.25 nmol, icv) alone had no effect, although a high dose (25 nmol, icv) increased Tb. It was hypothesized that the hyperthermia induced by nor-BNI was the result of the antagonist blocking the kappa opioid receptor and releasing its inhibition of mu opioid receptor activity. To determine whether the Tb increase caused by nor-BNI was a mu receptor-mediated effect, we administered the selective mu antagonist CTAP (1.25 nmol, icv) 15 min after nor-BNI (25 nmol, icv) and measured rectal Tb in unrestrained rats. CTAP significantly antagonized the Tb increase induced by icv injection of nor-BNI. Injection of 5 or 10 nmol of CTAP alone significantly decreased the Tb, and 1.25 nmol of nor-BNI blocked that effect, indicating that the CTAP-induced hypothermia was kappa-mediated. The findings strongly suggest that mu antagonists, in blocking the basal hyperthermia mediated by mu receptors, can unmask the endogenous kappa receptor-mediated hypothermia, and that there is a tonic balance between mu and kappa opioid receptors that serves as a homeostatic mechanism for maintaining Tb. PMID- 16257422 TI - Effects of hydrogen sulfide on hypoxic pulmonary vascular structural remodeling. AB - To study the role of hydrogen sulfide (H2S) in hypoxic pulmonary vascular structural remodeling (HPVSR), a total of 24 Wistar rats were randomly divided into three groups: control group (n = 8), hypoxia group (n = 8) and hypoxia with sodium hydrosulfide (hy + NaHS) group (n = 8). The mean pulmonary artery pressure (mPAP), plasma H2S and the percentage of muscularized arteries (MA), partially muscularized arteries (PMA) and nonmuscularized arteries (NMA) in small pulmonary vessels were measured. Collagen I and III, elastin, transforming growth factor beta3 (TGF-beta3), proliferative cell nuclear antigen (PCNA) and human urotensin II(U-II) expressions were detected by immunohistochemical assay. The mRNA expressions of procollagen I and III, matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of metalloproteinease-1 (TIMP-1) were detected by in situ hybridization. The results showed that NaHS significantly increased plasma H2S, decreased mPAP and the percentage of MA and PMA of small pulmonary vessels in rats under hypoxia. Meanwhile, NaHS inhibited the proliferation of pulmonary artery smooth muscle cells (PASMCs) represented by a decrease in the expressions of PCNA and human U-II in pulmonary artery wall. NaHS reduced the expression of collagen I and III, elastin and TGF-beta3 protein and decreased the expressions of procollagen I and III mRNA in pulmonary arteries of rats under hypoxia, but it did not impact the ratio of TIMP-1 mRNA to MMP-1mRNA in pulmonary arteries of rats under hypoxia. These data suggested that H2S played an important role in the development of HPVSR. PMID- 16257423 TI - Genetic immunization with LYVE-1 cDNA yields function-blocking antibodies against native protein. AB - LYVE-1 is a surface bound hyaluronic acid (HA) receptor that is preferentially expressed by lymphatic endothelial cells (LEC). cDNA encoding full-length human LYVE-1 was coated onto gold particles that were then delivered via helium assisted jet propulsion (gene gun) into the skin of Balb/C mice. LYVE-1 antisera, but not control pre-immune sera, recognized LYVE-1-transfected 293T cells by flow cytometry. While 40-70% of cultured human dermal microvascular endothelial cells (HMEC) were positive for LYVE-1 staining, human lung microvascular endothelial cells (LMEC) were negative. LYVE-1 antisera was used to effectively separate HMEC into LYVE-1 (hi) and LYVE-1(lo) populations that were enriched or depleted, respectively, for podoplanin, another LEC marker. By immunohistochemistry, LYVE-1 antisera detected CD31(lo) podoplanin(hi) lymphatic channels in normal and psoriatic human skin as well as in human tonsil. LYVE-1 antisera also blocked binding of FITC-labeled HA to HMEC (but not LMEC), demonstrating that these antibodies recognized regions of LYVE-1 required for HA binding. In summary, gene gun-assisted delivery of cDNA encoding LYVE-1 into skin resulted in reliable production of antisera that specifically and functionally recognized native LYVE 1 protein. PMID- 16257425 TI - Caveats in interpreting poison control centre data in spider bite epidemiology studies. PMID- 16257426 TI - High prevalence of hepatitis E antibodies in pregnant Egyptian women. AB - The epidemiology of hepatitis E virus (HEV), an enterically-transmitted cause of acute viral hepatitis (AVH), is not fully understood. During outbreaks on the Indian subcontinent and elsewhere, HEV causes severe AVH with mortality rates around 20% during pregnancy. In Egypt, where prevalence of HEV antibodies (anti HEV) in rural communities is very high, severe HEV-caused AVH in pregnant women has not been reported. This study examined a cohort of 2,428 pregnant women in the Nile Delta to assess prevalence of, and risk factors for, anti-HEV and correlated these with history of liver disease. Anti-HEV prevalence was 84.3%. Several risk factors associated with anti-HEV included older age, many siblings, not using soap to wash produce and frequent contact with cats. History of jaundice and liver disease was rare and not increased in those having anti-HEV. Our results confirm Egypt's high HEV endemicity and show that almost all women of childbearing age in these communities had prior HEV exposures without a history of liver disease. Reasons for the lack of clinical hepatitis remain unclear but could be the result of early childhood HEV exposures, producing long-lasting immunity and/or modify subsequent responses to exposure. Alternatively, the predominant HEV strain(s) in Egypt are less virulent than those in South Asia. PMID- 16257427 TI - Hepatitis E antibody seroconversion without disease in highly endemic rural Egyptian communities. AB - Hepatitis E virus (HEV) is enterically transmitted and causes self-limiting acute viral hepatitis (AVH) primarily in less developed countries. A prospective cohort study to assess incidence of, and risk factors for, seroconversion to HEV (anti HEV) was conducted in two Egyptian villages with a 67.7% anti-HEV prevalence. Nine hundred and nineteen villagers who were initially anti-HEV-negative were followed for 10.7 months. Thirty-four (3.7%) had strong anti-HEV serologic responses at follow-up giving an estimated anti-HEV incidence of 41.6/1,000 person-years. No significant associations were found between anti-HEV seroincidence and demographic and socioeconomic factors, source of water, household plumbing or sanitation, hand and vegetable washing, ownership of animals, jaundice and many other variables. None of the seroconverting subjects gave a history compatible with AVH during the interval. We hypothesize that both zoonotic and anthroponotic transmission of avirulent (possibly genotype-3) HEV is occurring extensively in these rural villages. An alternative explanation for the lack of morbidity among anti-HEV incident cases could be initial asymptomatic infections occur during early childhood with subsequent antibody titer boosting without illness upon re-exposure to the virus. PMID- 16257428 TI - Inference on microsatellite mutation processes in the invasive mite, Varroa destructor, using reversible jump Markov chain Monte Carlo. AB - Varroa destructor is a parasitic mite of the Eastern honeybee Apis cerana. Fifty years ago, two distinct evolutionary lineages (Korean and Japanese) invaded the Western honeybee Apis mellifera. This haplo-diploid parasite species reproduces mainly through brother-sister matings, a system which largely favors the fixation of new mutations. In a worldwide sample of 225 individuals from 21 locations collected on Western honeybees and analyzed at 19 microsatellite loci, a series of de novo mutations was observed. Using historical data concerning the invasion, this original biological system has been exploited to compare three mutation models with allele size constraints for microsatellite markers: stepwise (SMM) and generalized (GSM) mutation models, and a model with mutation rate increasing exponentially with microsatellite length (ESM). Posterior probabilities of the three models have been estimated for each locus individually using reversible jump Markov Chain Monte Carlo. The relative support of each model varies widely among loci, but the GSM is the only model that always receives at least 9% support, whatever the locus. The analysis also provides robust estimates of mutation parameters for each locus and of the divergence time of the two invasive lineages (67,000 generations with a 90% credibility interval of 35,000-174,000). With an average of 10 generations per year, this divergence time fits with the last post-glacial Korea-Japan land separation. PMID- 16257429 TI - Mating system and demographic constraints on the opportunity for sexual selection. AB - In monogamous systems the fitness difference between males due to competition for mates is limited to one female. This constraint presumably impedes the action of sexual selection relative to polygynous systems. In this paper, we use formal selection theory to show how population size and the adult sex ratio constrain the force of sexual selection and phenotypic evolution under monogamy and polygyny. The force of sexual selection is ultimately constrained by the number of males in a population and the theoretical limit to the rate of male phenotypic evolution is realized if a single male mates with one or many females. These results imply that the force of sexual selection is not strictly constrained by monogamy. The constraint on female phenotypic evolution is typically higher than the constraint on males under polygyny and similar to selection on males in monogamous systems. The sexual asymmetry in the force of selection under polygyny -not necessarily weak sexual selection on males of monogamous systems--may explain the prominence of sexual dimorphism in polygynous systems. PMID- 16257430 TI - Aryl hydrocarbon receptor-independent activation of estrogen receptor-dependent transcription by 3-methylcholanthrene. AB - Aryl hydrocarbon receptor (AhR) is a ligand-activated transcription factor that stimulates transcription directed by xenobiotic response elements upstream of target genes. Recently, AhR ligands were reported to induce formation of an AhR estrogen receptor (ER) complex, which can bind to estrogen response elements (EREs) and stimulate transcription of ER target genes. Presently, we investigate the effect of the AhR ligands 3-methylcholanthrene (3MC), 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) and 3,3',4,4',5-pentachlorobiphenyl (BZ126) on ERE-regulated luciferase reporter activity and endogenous ER target gene expression. In MCF-7 human breast cancer cells, 3MC induced transcription of ER reporter genes containing native promoter sequences of the ER-responsive genes complement 3 and pS2 and heterologous promoters regulated by isolated EREs. Dose response studies revealed that the concentration of 3MC required to half maximally activate transcription (EC(50)) was >100-fold higher for an ER reporter (27-57 muM) than for an AhR reporter (86-250 nM) in both MCF-7 cells and in human endometrial cancer Ishikawa cells. 3MC also stimulated expression of the endogenous ER target genes amphiregulin, cathepsin D and progesterone receptor, albeit to a much lower extent than was achieved following stimulation with 17beta estradiol. In Ishikawa cells, 3MC, but not BZ126 or TCDD, stimulated ERalpha dependent reporter activity but did not induce expression of endogenous ER target genes. Finally, studies carried out in the AhR-positive rat hepatoma cell line 5L and the AhR-deficient variant BP8 demonstrated that ER reporter activity could be induced by 3MC in a manner that was independent of AhR and thus distinct from the AhR-ER 'hijacking' mechanism described recently. 3MC may thus elicit estrogenic activity by multiple mechanisms. PMID- 16257432 TI - The human papillomavirus type 11 E1--E4 protein is a transglutaminase 3 substrate and induces abnormalities of the cornified cell envelope. AB - The human papillomavirus (HPV) E1--E4 protein is detected in the cytoplasm of differentiated keratinocytes, near the cornified cell envelope. HPV does not induce lysis of the infected keratinocyte, and the normally durable cornified cell envelope that forms during keratinocyte differentiation would seemingly inhibit viral egress. HPV infection induces abnormalities of the cornified cell envelope, but the exact mechanisms involved are not well understood. We tested whether the HPV 11 E1--E4 protein, which co-localizes the cell envelope and co purifies with cell envelope fragments, could serve as an in vitro substrate for transglutaminases. We found evidence of E1--E4 cross-linking by endogenous transglutaminases in an in situ assay using frozen sections of human foreskin, and in addition, E1--E4 protein was cross-linked by recombinant transglutaminase 3 (but not transglutaminase 1) in an in vitro cross-linking assay. We also tested whether expression of E1--E4 in differentiated keratinocytes would induce morphologic alterations of cornified cell envelopes. Differentiated keratinocytes expressing E1--E4 were disorganized and pleomorphic compared to control cells, and cell envelopes purified from E1--E4-expressing cells were small, fragmented, and rough bordered compared to the round, smooth bordered cell envelopes from control cells. We conclude from these in vitro experiments that the E1--E4 protein is cross-linked by transglutaminase 3, and that E1--E4 expression in differentiated keratinocytes induces morphologic abnormalities of the cornified cell envelope. PMID- 16257431 TI - Purification, primary structures and evolution of coagulant proteases from Deinagkistrodon actus venom. AB - Deinagkistrodon (formerly Agkistrodon) actus (Taiwan) snake venom was found to contain at least seven closely related coagulant proteases. One of them, named actibin, was purified to homogeneity by means of four chromatographic steps. Actibin acted on fibrinogen to form fibrin clots with extremely high specific activity of 1,630 NIH units/mg and preferentially released fibrinopeptide A. Actibin was an acidic glycoprotein (pI 3.4) with molecular weight of 41,000, which was reduced to 28,800 after deglycosylation with N-glycanase. The k(cat)/K(m) values of actibin for hydrolysis of tosyl-l-arginine methyl ester and benzoyl-l-arginine p-nitroanilide were one-third to a half those for thrombin, reflecting a high potency of actibin in fibrinogen clotting. The amidase activities of actibin and its family proteases were inhibited by 3,4 dichloroisocoumarin, a serine protease inhibitor, indicating that actibin and its family proteases are serine proteases. Four cDNAs, named DaP1 and DaP7-DaP9, encoding D. actus coagulant proteases were cloned. All cDNAs contain an open reading frame of 780 bp coding for 260 amino acid residues, including a signal peptide of 24 amino acid residues. Their amino acid sequences predicted are highly homologous to one another with one to five amino acid substitutions. When four D. actus protease cDNAs were compared with the cDNAs coding for Trimeresurus flavoviridis and T. gramineus venom serine proteases, accelerated evolution was clearly observed. Similarity of the nucleotide sequences of four D. actus protease cDNAs with no synonymous and one to five nonsynonymous substitutions seems not to be in direct conformity with accelerated evolution. This possibly suggests that they have evolved to a similar direction to enhance their clotting activity rather than to produce other physiological activities. PMID- 16257433 TI - Macrobenthos community structural changes off Cesenatico coast (Emilia Romagna, Northern Adriatic), a six-year monitoring programme. AB - Soft bottom macrobenthos at a station located off Cesenatico (Emilia Romagna, Northern Adriatic Sea) was investigated seasonally for six years from July 1996 to July 2002. Species composition and abundance of the community have been studied in relation to fluctuation in the water environment parameters, sediment texture patterns and mucilage, that occurred mainly in the water column at the study site. Three major Po river flow peaks occurred in November 1996, October 2000 and May 2002; after these events the community was reduced to minimum abundance values (total density<2000 individuals m(-2)). In the period between the first two episodes the river discharge remained rather low and conditions of increased salinity, lower nutrients and chl a and good oxygen saturation were experienced. The fossorial Crustacean Ampelisca diadema became dominant in the community between the first two river flow events, reaching maximum density of 10,200 individuals m(-2) and substituting the bivalve Corbula gibba, indicator of sediment instability. Species richness increased in the same period. The role of Ampelisca as a facilitator in structuring the community is discussed. Corbula gibba never recovered to initial densities, apart from an abundance peak that occurred in the summer of 2000. Faunal composition seemed to evolve slowly towards a higher degree of structural complexity (positive trend in diversity and evenness index). In the study site near-bottom mucilage events occurred in the summers of 1997, 1998, 2002; they appeared uncorrelated with the observed changes in the community structure. Multivariate analysis of community structure (MDS, ANOSIM) illustrates that community changes in this station are driven mainly by hydrographical conditions influencing sediment texture patterns and trophic resources for the benthos. PMID- 16257434 TI - Chemical characterization of different typologies of mucilaginous aggregates in the Northern Adriatic Sea. AB - The chemical composition of mucilage aggregates found during summer 2000, 2001 and 2002 in the North Adriatic Sea depends on the nature of the organic matter during aggregation, on the environmental conditions of the site of formation and on the transformations during ageing. The mucilages were composed of organic matter, together with a significant inorganic fraction. Elemental analysis revealed 12.5-32.2% of organic carbon, 0-7.3% of inorganic carbon and 1.0-3.7% of nitrogen. The C(org)/N ratios of most aggregates were between 7.5 and 12.6, values close to those found in the suspended matter; higher ratios were found in large-size (>5 m) aggregates which are probably older. The content of carbohydrates and proteins determined in the aggregates, respectively, 15.4+/ 8.9% and 7.9+/-4.8%, w/w, showed a prevalence of carbohydrates over proteins. Neutral carbohydrate analysis of purified polysaccharides from mucilage samples showed very similar signatures with high relative abundance of galactose and glucose. Humic, fulvic and humin substances extracted from the mucilages constitute an important fraction of the organic matter in the aggregates. The humin (a fraction insoluble in acidic and basic media) was present in all mucilage samples, indicating the refractory nature of a part of the organic matter in the mucilage. The iron and calcium could play a role during the aggregation process to form a complex with polysaccharides and humic fractions. The C(org)/N ratio 10+/-2 found in the humic acids extracted from the Adriatic aggregates disclosed a marine origin. The low phosphorus content and the high C(org)/P ratio found in the aggregates might depend from high bacteria activity or from the aggregation of organic fractions depleted of phosphorus. The principal inorganic species contained aluminium and silicon, part of which was of biogenic origin and was more significant in the offshore mucilage aggregates than in the coastal ones. The Si(biog)/C(org) ratio showed that diatoms were always present in the aggregates, although it cannot be established whether these are the producers or these develop within the aggregates. PMID- 16257435 TI - Inter-annual variations of planktonic food webs in the northern Adriatic Sea. AB - The temporal dynamics of microphytoplankton, microzooplankton and mesozooplankton were monitored over 37 months in the Adriatic Sea in order to identify alterations in the plankton structures, which can lead to, or enhance the production of macro-aggregates, that affected the entire northern basin in summers 2000 and 2002, and to assess any negative effects of mucilage on plankton temporal patterns. Samples were collected monthly, from June 1999 to July 2002, on three transects at 9 stations across the northern and central Adriatic Sea. Besides the high year-to-year variations in abundances and taxonomical composition, plankton communities only showed a clear seasonal succession during 2001, when since April a grazing food web developed and was able to control large sized phytoplankton increase. In spring-summer 2000 and 2002 consumer abundances remained quite low and the dominant mesozooplankton summer species (Penilia avirostris) did not reach its usual summer maximum. The lack of an efficient top control was more evident on the northernmost transect, where generally grazing food web prevails over the microbial one. A large part of the microphytoplankton blooms, although not particularly intense, was exported to the bottom in the particulate phase, where it was processed by bacteria, enhancing the production of refractory dissolved material. PMID- 16257436 TI - The physicochemical characterisation of microscopic airborne particles in south Wales: a review of the locations and methodologies. AB - As part of the NERC-URGENT thematic programme, research was undertaken into the physicochemistry and bioreactivity of microscopic airborne particulate matter in south Wales. This paper reviews the collecting and characterisation methods used in the research; some of the results obtained are shown as examples. Four main collecting locations were chosen: Cardiff (urban); Port Talbot (urban/industrial); Park Slip West coal opencast pit (industrial/rural); the Black Mountains (rural/background). Collections initially used a 30-l/min Negretti PM10 filter collection system, however in the later stages of the project increased use was made of a 1100-l/min impaction system (nicknamed the super-sucker). This latter device was developed at Harvard University USA, however was adapted and optimised at Cardiff University. Methods for the extraction of PM10 off polycarbonate filters and polyurethane substrates were developed, with particular attention being paid to minimise physical or chemical changes during the extraction, and the extracts being in an appropriate state for bioreactivity assessment. Physicochemical characterisation of the PM10 included the empirical measurement of shape and size using electron microscopy and semi automated image analysis. The determinations of the water-soluble and -insoluble chemical components were undertaken by ion chromatography and inductive coupled plasma-mass spectrometry. The bioreactivity of south Wales airborne particles is not covered by this review. PMID- 16257437 TI - The influence of forestry activity on the structure of dissolved organic matter in lakes: implications for mercury photoreactions. AB - It is well known that dissolved organic matter (DOM) increases in lakes associated with forestry activity but characterization of the DOM structure is incomplete. Twenty-three lakes with a wide range of forestry activities located in central Quebec, Canada were sampled and analyzed for dissolved organic carbon (DOC) concentration, DOC fluorescence, and ultra violet-visible (UV-VIS) absorption spectra. The results show that DOC increases (as does the associated DOC fluorescence) with increased logging (slope=0.122, r2=0.581, p<0.001; and slope=0.283, r2=0.308, p<0.01, respectively) in the 23 lakes sampled however, the aromaticity of the DOM does not change with changes in logging (as found by UV VIS ratios, absorbance slope in the UV region, and DOC normalized fluorescence (slope=1.42x10(-2), r2=0.331, p<0.01). The DOM from four of these lakes was concentrated using reverse osmosis (RO) followed by freeze-drying. The structures of the concentrated dissolved organic matter (DOM) samples were analyzed using X ray analysis of near edge structures (XANES), X-ray diffraction (XRD), and 13C solid-state nuclear magnetic resonance (13C NMR) analysis. XANES analysis of functional groups in the four concentrated samples shows that there are significant differences in reduced sulphur between the samples, however there was no clear relationship with forestry activity in the associated catchment. XRD data showed the presence of amorphous sulphide minerals associated with the DOM concentrate that may be important sites for mercury binding. The 13C NMR spectra of these samples show that the percentage of carbon present in carboxylic functional groups increases with increasing logging. Such structures are important for binding photo-reducible mercury and their presence may limit mercury photo-reduction and volatilization. We propose a mechanism by which increased logging leads to increased carboxylic groups in DOM and thereby increased weak binding of photo-reducible mercury. These results, in part, explain the decrease in dissolved gaseous mercury (DGM) production rates with increased logging found in our previous work. PMID- 16257438 TI - A numerical study of the interannual variability of the Adriatic Sea (2000-2002). AB - A free-surface, three-dimensional finite-difference numerical model based on the Princeton Ocean Model (POM) has been implemented in order to simulate the interannual variability of the Adriatic Sea circulation. The implementation makes use of an interactive surface momentum and heat flux computation that utilizes the European Centre for Medium-Range Weather Forecasts (ECMWF) 6-h analyses and the model predicted sea surface temperatures. The model is also nested at its open boundary with a coarse-resolution Mediterranean general circulation model, utilizing the same surface forcing functions. The simulation and analysis period spans 3 years (1 Jan 2000 to 31 Dec 2002) coinciding with the "Mucilage in the Adriatic and the Tyrrhenian" (MAT) Project monitoring activities. Model results for the simulated years show a strong interannual variability of the basin averaged proprieties and circulation patterns, linked to the atmospheric forcing variability and the Po river runoff. In particular, the years 2000 and 2002 are characterized by a weak surface cooling (with respect to the climatological value) and well-marked spring and autumn river runoff maxima. Conversely, 2001 is characterized by stronger wind and heat (autumn cooling) forcings but no river runoff autumn peak, even though the total amount of water inflow during winter and spring is sustained. The circulation is characterized by similar patterns in 2000 and 2002 but very different structures in 2001. During the latter, deep water is not formed in the northern Adriatic. A comparison with the observed data shows that the major model deficiencies are connected to the low salinity of the waters, probably connected to the missed inflow of salty Ionian waters of Aegean origin and to the numerical overestimation of the vertical mixing processes. PMID- 16257439 TI - Vitamins and minerals that affect hemostasis and antithrombotic therapies. PMID- 16257440 TI - FDG-PET in the detection of recurrence of uterine cervical carcinoma following radiation therapy--tumor volume and FDG uptake value. AB - PURPOSE: We evaluated the use of positron emission tomography (PET) with fluorine 18-labeled fluoro-2-deoxy-d-glucose (FDG) in follow-up study after radiation therapy in patients with uterine cervical carcinoma. MATERIALS AND METHODS: Thirty-two studies in 25 patients were reviewed. Twenty patients were treated with external beam irradiation and intracavitary brachytherapy, and five with irradiation following initial surgery. Time from initial treatment to FDG-PET was 23.3 (5.2-88.0) months. Rationale for FDG-PET was the presence of symptoms in 6 patients, abnormal serum tumor marker values in 13, abnormal lesions on other diagnostic imaging modalities in 19, and patient request in 2. On visualization of a lesion, the maximum standardized uptake value (maxSUV) of the lesion was calculated, and values over 2.0 were classified as FDG-positive. Maximum tumor diameter and tumor volume in the corresponding disease were estimated by computed tomography (CT) or magnetic resonance imaging (MRI). RESULTS: Sensitivity and specificity of FDG-PET in the detection of recurrent disease were 91.5% (43/47) and 57.1% (4/7), respectively. Four false-negative findings were seen for small lung metastases having a volume less than 1 cm3. Three false-positive cases were a localized pneumonitis, a benign pubic bone fracture, and a fibrosis after interstitial brachytherapy. Sensitivity for extrapelvic lymph node metastases was extremely high (100%); in contrast, sensitivity and specificity for lung and bone lesions were 75.0% (12/16) and 33.3% (1/3), respectively. Regarding tumor volume measurement, good correlation between maxSUV on FDG-PET and tumor volume was obtained (lung metastases, P = 0.03; extrapelvic nodes, P < 0.0001). Within this study, all corresponding lesions over 1 cm3 showed a maxSUV value greater than 2.0. CONCLUSION: FDG-PET is a useful tool for the detection of extrapelvic lesions during the follow-up period after radiation therapy for cervical cancer. This study suggests that FDG uptake is associated with tumor volume, and FDG-PET has limitations in the detection of lesions less than 1 cm3 or microscopic disease. Careful diagnostic agreement between PET and CT/MRI for positive but benign lesions, such as inflammation and bone fracture, remains important. PMID- 16257441 TI - Trust in health care: seeing it from the other side. PMID- 16257443 TI - Post-hatching development of the porcine and bovine embryo--defining criteria for expected development in vivo and in vitro. AB - Particular attention has been paid to the pre-hatching period of embryonic development although blastocyst development is a poor indicator of embryo viability. Post-hatching embryonic development in vitro would allow for establishment of more accurate tools for evaluating developmental potential without the need for transfer to recipient animals. Such a system would require (1) definition of milestones of expected post-hatching embryonic development in vivo; and (2) development of adequate culture systems. We propose a stereomicroscopical staging system for post-hatching embryos defining the following stages: (1) Expanded hatched blastocyst stage where the embryo presents an inner cell mass (ICM) covered by trophoblast. (2) Pre-streak stage 1 where the embryonic disc is formed. (3) Pre-streak stage 2 where a crescent-shaped thickening of the caudal portion of the embryonic disk appears. (4) Primitive streak stage where the primitive streak has developed as an axis of cell ingression of cells for meso- and endoderm formation. (5) Neural groove stage where the neural groove is developing from the rostral pole of the embryo along with a proportional shortening of the primitive streak; and (6) Somite stage(s) where paraxial mesoderm gradually condensates to form somites. Post-hatching development of bovine embryos in vitro is compromised and although hatching occurs and elongation can be physically provoked by culture in agarose tunnels, the embryonic disk characterizing the pre-streak stage 1 is never established. Thus, particular focus should be placed on establishing culture conditions that support at least some of the above-mentioned critical phases of development that in vivo occur within the initial two (pig) to three (cattle) weeks. PMID- 16257444 TI - Surface functionalization of degradable polymers by covalent grafting. AB - With a new non-destructive and solvent-free photografting technique, N vinylpyrrolidone was covalently grafted onto the surfaces of degradable polymers; poly(l-lactide), poly(epsilon-caprolactone), poly(lactide-co-glycolide), and poly(trimethylene carbonate). The modified surfaces were characterized by XPS, ATR-FTIR, SEM, and cell growth tests. The wettability was markedly improved, as static contact angles changed from about 80 degrees for the pristine substrates to around 30 degrees after 30min of grafting. Well-defined surface topographies, such as micro-patterns, are preserved in the process since the graft layers are thin. The biological response, measured as cytotoxicity, showed that the modified films provide good substrates, comparable with optimized cell culture plastics, for the adhesion and proliferation of normal human keratinocytes and skin fibroblasts. PMID- 16257446 TI - Evolutionary conservation of alternative activation of macrophages: structural and functional characterization of arginase 1 and 2 in carp (Cyprinus carpio L.). AB - Classically activated macrophages (caMF) play an important role in type-I immune responses and alternatively activated macrophages (aaMF) function in type-II immune responses. While the classical activation of fish macrophages has been well described, the existence of aaMF has not yet been described for teleosts. Arginase is the characteristic enzyme in aaMF and two isoforms have been described for mammals. To study the presence of aaMF in a primitive vertebrate species we cloned arginase 1 and 2 cDNA of common carp. Carp arginase 1 is a 340 aa protein with 63% aa sequence identity to human arginase 1. Carp arginase 2 is a 347 aa protein with 63% aa sequence identity to human arginase 2. Three highly homologous arginase 2 genes were found, each showing only single non-synonymous substitutions. Basal arginase 1 expression is mainly found in carp mid kidney. In contrast, arginase 2 was expressed in all organs examined with the highest basal gene expression in liver. Cultured carp head kidney-derived macrophages were used to study aaMF in vitro. Carp macrophages showed significant arginase activity which could be induced by dibutyryl cyclic adenosine mono phosphate (cAMP) and specifically inhibited by NG-hydroxy-L-arginine (NOHA). At the gene level, arginase 2 gene expression was upregulated by cAMP stimulation, while arginase 1 gene expression was not influenced. LPS stimulation did not alter the arginase 1 or 2 expression, inducible nitric oxide synthase (iNOS) expression was, however, upregulated. This expression of iNOS was used as a measure of classical activation of carp macrophages. Thus, in contrast to mammals, fish arginase 2 and not arginase 1 is differentially regulated and likely involved in the alternative activation of fish macrophages. Our data suggest there may be an evolutionary conservation of the presence of aaMF down to teleost fish. PMID- 16257445 TI - Mode of mitochondrial Ca2+ clearance and its influence on secretory responses in stimulated chromaffin cells. AB - To study the role of mitochondrial Ca(2+) clearance in stimulated cells, changes in free Ca(2+) concentration in the cytosol, [Ca(2+)](c) and that in mitochondria, [Ca(2+)](m) along with secretory responses were observed using chromaffin cells co-loaded with Fura-2 and Rhod-2 in the perfused rat adrenal medulla. When the cells were stimulated with 40 mM K(+) in the perfusate, the duration of [Ca(2+)](m) response markedly increased with prolongation of the stimulation period, exhibiting a mean half-decay time of 21 min with 30s stimulation, whereas its amplitude was not altered with stimulations of 10-30s. A computer simulation analysis showed that such a mode of [Ca(2+)](m) response can be produced if excess Ca(2+) taken up by mitochondria precipitates as calcium phosphate (Pi) salt. In the presence of 5 microM rotenone plus 10 microM oligomycin, a decrease in the duration of [Ca(2+)](m) response and a slight but significant increase (24%) in the secretory response to 30s stimulation with 40 mM K(+) were observed. Simulation analyses suggested that this effect of rotenone may be due to reduction in mitochondrial Ca(2+) uptake induced by rotenone elicited partial depolarization of the mitochondrial membrane potential. In chromaffin cells transsynaptically stimulated through the splanchnic nerve, the intensity of NAD(P)H autofluorescence changed with time courses similar to those of [Ca(2+)](m) responses. The temporal profiles of those two responses were prolonged in a similar manner by application of an inhibitor of mitochondrial Na(+)/Ca(2+) exchanger, CGP37157. Thus, due to the unique Ca(2+) buffering mechanism, [Ca(2+)](m) responses associated with massive mitochondrial Ca(2+) uptake may occur within a limited concentration range in which Ca(2+)-sensitive dehydrogenases are activated to control the mitochondrial redox state in stimulated chromaffin cells. PMID- 16257447 TI - Cloning and sequence analysis of the beta2-microglobulin transcript from flounder, Paralichthys olivaceous. AB - Beta2-microglobulin (beta2M) is a protein found free-form in the serum or on the cell surface non-covalently associated with the alpha-chain of the class I major histocompatibility (MHC-I) complex. The full-length cDNA containing beta2M was cloned from flounder, Paralichthys olivaceous. The transcript consists of 1610 nucleotides (nts), including an open reading frame (ORF) of 384 nts encoding a polypeptide of 128 amino acids. The amino acid sequence of beta2M in flounder is 59, 57, 56, and 48% conserved in catfish, rainbow trout, zebrafish, and humans, respectively. Genomic Southern hybridization suggested the presence of a single copy of beta2M in the flounder genome, and reverse transcription-polymerase chain reaction (RT-PCR) and Northern blot analysis detected the beta2M transcript in the head kidney, spleen, body kidney, liver, and muscle tissues of the flounder. PCR amplification and sequence analysis revealed the lack of an intron in the beta2M gene. The phylogenetic analysis confirmed the evolutionary diversion of the beta2M protein among warm-blooded vertebrates and fish, and the separation between freshwater and seawater fish. PMID- 16257449 TI - Pharmacological profiles of cloned mammalian P2Y-receptor subtypes. AB - Membrane-bound P2-receptors mediate the actions of extracellular nucleotides in cell-to-cell signalling. P2X-receptors are ligand-gated ion channels, whereas P2Y receptors belong to the superfamily of G-protein-coupled receptors (GPCRs). So far, the P2Y family is composed out of 8 human subtypes that have been cloned and functionally defined; species orthologues have been found in many vertebrates. P2Y1-, P2Y2-, P2Y4-, P2Y6-, and P2Y11-receptors all couple to stimulation of phospholipase C. The P2Y11-receptor mediates in addition a stimulation of adenylate cyclase. In contrast, activation of the P2Y12-, P2Y13-, and P2Y14 receptors causes an inhibition of adenylate cyclase activity. The expression of P2Y1-receptors is widespread. The receptor is involved in blood platelet aggregation, vasodilatation and neuromodulation. It is activated by ADP and ADP analogues including 2-methylthio-ADP (2-MeSADP). 2'-Deoxy-N6-methyladenosine 3',5'-bisphosphate (MRS2179) and 2-chloro-N6-methyl-(N)-methanocarba-2' deoxyadenosine 3',5'-bisphosphate (MRS2279) are potent and selective antagonists. P2Y2 transcripts are abundantly distributed. One important example for its functional role is the control of chloride ion fluxes in airway epithelia. The P2Y2-receptor is activated by UTP and ATP and blocked by suramin. The P2Y2 agonist diquafosol is used for the treatment of the dry eye disease. P2Y4 receptors are expressed in the placenta and in epithelia. The human P2Y4-receptor has a strong preference for UTP as agonist, whereas the rat P2Y4-receptor is activated about equally by UTP and ATP. The P2Y4-receptor is not blocked by suramin. The P2Y6-receptor has a widespread distribution including heart, blood vessels, and brain. The receptor prefers UDP as agonist and is selectively blocked by 1,2-di-(4-isothiocyanatophenyl)ethane (MRS2567). The P2Y11-receptor may play a role in the differentiation of immunocytes. The human P2Y11-receptor is activated by ATP as naturally occurring agonist and it is blocked by suramin and reactive blue 2 (RB2). The P2Y12-receptor plays a crucial role in platelet aggregation as well as in inhibition of neuronal cells. It is activated by ADP and very potently by 2-methylthio-ADP. Nucleotide antagonists including N6-(2 methylthioethyl)-2-(3,3,3-trifluoropropylthio)-beta,gamma-dichloromethylene-ATP (=cangrelor; AR-C69931MX), the nucleoside analogue AZD6140, as well as active metabolites of the thienopyridine compounds clopidogrel and prasugrel block the receptor. These P2Y12-antagonists are used in pharmacotherapy to inhibit platelet aggregation. The P2Y13-receptor is expressed in immunocytes and neuronal cells and is again activated by ADP and 2-methylthio-ADP. The 2-chloro-5-nitro pyridoxal-phosphate analogue 6-(2'-chloro-5'-nitro-azophenyl)-pyridoxal-alpha5 phosphate (MRS2211) is a selective antagonist. mRNA encoding for the human P2Y14 receptor is found in many tissues. However, a physiological role of the receptor has not yet been established. UDP-glucose and related analogues act as agonists; antagonists are not known. Finally, UDP has been reported to act on receptors for cysteinyl leukotrienes as an additional agonist--indicating a dual agonist specificity of these receptors. PMID- 16257448 TI - Synthesis, characterization, DNA-binding and photocleavage of complexes [Ru(phen)2(6-OH-dppz)]2+ and [Ru(phen)2(6-NO2-dppz)]2+. AB - Two new complexes, ([Ru(phen)(2)(6-OH-dppz)](2+)) (1) and ([Ru(phen)(2)(6-NO(2) dppz)](2+)) (2) (phen=1,10-phenanthroline; 6-OH-dppz=6-hydroxyl-dipyrido[3,2 a:2',3'-c]phenazine; 6-NO(2)-dppz=6-nitro-dipyrido[3,2-a:2',3'-c]phenazine), have been synthesized and characterized by elemental analysis, ES-MS (electrospray mass spectra), (1)H NMR, UV-Vis (UV-visible) and CV (cyclic voltammetry). The DNA binding behaviors of both complexes have been studied by spectroscopic methods and viscosity measurements. The results indicate that the two complexes all bind to calf thymus DNA (CT-DNA) in an intercalative mode, and the DNA-binding affinity of complex 2 is greater than that of complex 1. In addition, complex 1 can promote photocleavage of pBR322 DNA upon irradiation, whereas complex 2 can promote cleavage of pBR322 DNA both upon irradiation and in the dark, with more efficient cleavage occurring upon irradiation. Theoretical studies for these complexes have been also carried out with the density functional theory (DFT) method. The difference in the DNA-binding behaviors of the two complexes can be reasonably explained by the DFT calculations. PMID- 16257450 TI - Noninvasive method of immortalized neural stem-like cell transplantation in an experimental model of Huntington's disease. AB - A loss of neostriatal neurons is a characteristic of Huntington's disease (HD), and neural tissue transplantation has been performed directly into the striatum. Since the neural stem cells have ability to migrate into the lesion site, we administered immortalized neural stem-like cells (NSC) into the ventricle or via a tail vein following unilateral intrastriatal quinolinic acid lesioning in Sprague-Dawley rats. To identify transplanted NSC, cells were encoded with lac Z and beta-galactosidase histochemistry was performed. lac Z+ cells were detected in the lesioned striatum but tissue damage or tumor formation was not observed. This study shows that NSC migrate into the striatum, either from ventricle or from the systemic circulation, providing less invasive routes for stem cell application in HD. PMID- 16257451 TI - Confinement and other psychosocial factors in perinatal depression: a transcultural study in Singapore. AB - BACKGROUND: To investigate the prevalence, socio-cultural and psychosocial risk factors for perinatal depression in Singaporean women. METHOD: A prospective cohort of 559 women was interviewed antenatally and at six weeks' postpartum at a tertiary hospital. Women were interviewed for diagnosis of depression using a two stage design, with a screening questionnaire and diagnostic interview. RESULTS: Postnatally, a negative confinement experience was associated with depression. Other independent factors included poor emotional support, a past history of depression, unplanned pregnancy and perceived potential conflicts with relatives over childcare antenatally and dissatisfaction, poor instrumental support postnatally. The prevalence of depression antenatally and postnatally was 12.2% and 6.8%, respectively. LIMITATIONS: Measures of satisfaction with social support were based on self-report; there were high dropout rates at six weeks' postpartum; and other modulating social factors such as pre-existing interpersonal conflicts were not studied. CONCLUSIONS: Perinatal depression in Singaporean women is common. Contrary to expectations, a negative 'confinement' experience is a significant risk factor for postnatal depression, and is not universally welcomed by women. Depression is modulated by dissimilar sets of psychosocial factors antenatally and postnatally. PMID- 16257452 TI - Marijuana for cholera therapy. PMID- 16257453 TI - Apology received. The circadian clock in mammals. PMID- 16257454 TI - Seasonal variation of the impact of a stressful procedure on open field behaviour and blood corticosterone in laboratory mice. AB - Behavioural and hormonal seasonal changes are well documented in various vertebrate species living in their natural environment but circannual variations that may occur in laboratory animals reared in standard conditions are poorly investigated. This study shows that, in laboratory mice, the effects of stress on behavioural inhibition, investigatory behaviour and blood concentration of corticosterone are seasonally dependent. No consistency was observed between the reactivity of biological structures controlling the hormonal response to stress and the behavioural activities investigated at every period of the year. During the spring time, stress, which elicited a decrease of investigatory behaviour (estimated by the walking time in an open field), increased behavioural inhibition (estimated by the percentage of walking in the central area of the open field) as well as the blood corticosterone concentration in laboratory mice. In autumn, stress had no significant effect on behaviour despite the great hormonal concentration increase. The results reveal that, at certain period of the year, a stressful procedure is unable to affect behavioural parameters in laboratory mice which were maintained in constant 12-h dark/12-h light cycle. The report constitutes a novel piece of information suggesting a potential role of the endogenous biological clock in the modulation of stress response in mammals. PMID- 16257455 TI - Repeated subchronic exposure to phencyclidine elicits excessive atypical grooming in rats. AB - Self-grooming in rodents is stereotypically sequenced and naturally occurs after arousal, novelty, or stress. Grooming expression and syntax resulting from stressful and appetitive conditions were assessed in male Long Evans rats treated daily with 10mg/kg of phencyclidine (PCP) for 15 days. Approximately 20 h after the 1st, the 8th, and/or the 15th injection, grooming was induced with water sprays, a loud sound, or smearing food. Behaviors expressed during the seconds or minutes that followed induction were videotaped and codified. Results showed that subchronic treatment with PCP amplified the grooming response in all stressful and appetitive conditions, but provoked a disorganization of grooming sequences only under the stressful, water condition. Thus, PCP enhanced grooming expression indiscriminately. However, this behavior had to serve both hygienic and stress managing purposes in order for chain sequencing to become disorganized as a consequence of drug treatment. These results suggest that the detailed examination of grooming expression and organization is an appropriate tool to measure stress-induced behavioral sensitization and motor functions in animal models of neuropsychiatric disorders such as schizophrenia. PMID- 16257456 TI - Pre-replication complex organization in the atypical DNA replication cycle of Plasmodium falciparum: characterization of the mini-chromosome maintenance (MCM) complex formation. AB - The overall organization of cell division in Plasmodium is unique compared to that observed in model organisms because DNA replicates more than once per cell cycle at several points of its life cycle. The sequencing of the Plasmodium genome has also revealed the apparent absence of many key components (e.g. Cdt1, DDK and Cdc45) of the eukaryotic cell cycle machinery that are responsible for the formation of the pre-replication complex (pre-RC). We have characterized the Plasmodium falciparum minichromosome maintenance complex (MCM) that plays a key role in the transition of pre-RC to the RC. Similar to other eukaryotes, the Plasmodium genome encodes six MCM subunits. Here, we show that expression levels of at least three of the PfMCM subunits, the homologues of MCM2, MCM6 and MCM7, change during the intraerythrocytic development cycle, peaking in schizont and decreasing in the ring and trophozoite stages. PfMCM2, 6 and 7 subunits interact with each other to form a developmentally regulated complex: these interactions are detectable in rings and schizonts, but not in trophozoites. PfMCM2, 6 and 7 subunits are localized in both cytosolic and nucleosolic fractions during all intraerythrocytic stages of P. falciparum development, with increased nuclear localization in schizonts. Only PfMCM6 is associated with the chromatin fraction at all stages of growth. No phosphorylation of PfMCM2, 6 and 7 was detected, but two as yet unidentified threonine-phosphosphorylated proteins were present in the complex, whose pattern of phosphorylation varied during parasite development. PMID- 16257457 TI - Cloning and characterization of a zebrafish Y2 receptor. AB - The NPY receptors belong to the superfamily of G-protein coupled receptors and in mammals this family has five members, named Y1, Y2, Y4, Y5, and Y6. In bony fish, four receptors have been identified, named Ya, Yb, Yc and Y7. Yb and Y7 arose prior to the split between ray-fined fishes and tetrapods and have been lost in mammals. Yc appeared as a copy of Yb in teleost fishes. Ya may be an ortholog of Y4, but surprisingly no unambiguous receptor ortholog to any of the mammalian subtypes has yet been identified in bony fishes. Here we present the cloning and pharmacological characterization of a Y2 receptor in zebrafish, Danio rerio. To date, this is the first Y2 receptor outside mammals and birds that has been characterized pharmacologically. Phylogenetic analysis and synteny confirmed that this receptor is orthologous to mammalian Y2. We show that the receptor is pharmacologically most similar to chicken Y2 which leads to the conclusion that Y2 has acquired several novel characteristics in mammals. Y2 from zebrafish binds very poorly to the Y2-specific antagonist BIIE0246. Our pharmacological characterization supports our previous conclusions regarding the binding pocket of BIIE0246 in the human Y2 receptor. PMID- 16257458 TI - Disruption of disulfide bond formation alters the trafficking of prothyrotropin releasing hormone (proTRH)-derived peptides. AB - Rat prothyrotropin releasing hormone (proTRH) is processed in the regulated secretory pathway (RSP) of neuroendocrine cells yielding five TRH peptides and several non-TRH peptides. It is not understood how these peptides are targeted to the RSP. We show here that a disulfide bond in the carboxy-terminus of proTRH plays an important role in the trafficking of this prohormone. Recombinant proTRH was observed to migrate faster on a native gel when treated with dithiothreitol (DTT) suggesting the presence of a disulfide bond. In vitro disulfide bond formation was prevented either by DTT treatment or by mutating cysteines 213 and 219 to glycines. In both cases the peptides derived from these mutants exhibited increased constitutive release and processing defects when expressed in AtT20 cells, a neuroendocrine cell line used in our prior studies on proTRH processing. Immunocytochemistry revealed that wild-type proTRH and mutant proTRH localized in a punctate pattern typical of proteins sorted to the regulated secretory pathway. These data suggest that the proposed disulfide bond of proTRH is involved in sorting of proTRH-derived peptides and in their retention within maturing secretory granules. This is the first evidence of structural motifs being important for the sorting of proTRH. PMID- 16257459 TI - Monte Carlo simulations of VEGF binding to cell surface receptors in vitro. AB - The vascular endothelial growth factor (VEGF) family binds multiple endothelial cell surface receptors. Our goal is to build comprehensive models of these interactions for the purpose of simulating angiogenesis. In view of low concentrations of growth factors in vivo and in vitro, stochastic modeling of molecular interactions may be necessary. Here, we compare Monte Carlo simulations of the stochastic binding of VEGF and two of its major receptors on cells in vitro to equivalent deterministic simulations. In the range of typical VEGF concentrations, the stochastic and deterministic models are in agreement. However, we observe significant variability in receptor binding, which may be linked to biological stochastic events, e.g., blood vessel sprout initiation. We study patches of cell surface of varying sizes to investigate spatial integration of the signal by the cell, which impacts directly the variability of binding, and find significant variability up to the single-cell level. Dimerization of VEGF receptors does not significantly alter the variability in ligand binding. A 'sliding window' approach demonstrated no reduction in the variability of binding by temporal integration. The variability is expected to be more prominent in in vivo situations where the number of ligand molecules available for binding is less. PMID- 16257460 TI - Cardiac sarcoidosis evaluated with gadolinium-enhanced magnetic resonance and contrast-enhanced 64-slice computed tomography. AB - Sarcoidosis is a multi-system granulomatous disorder of unknown etiology with symptomatic cardiac involvement in up to 7% of patients. The clinical features of sarcoid heart disease include congestive heart failure, arrhythmias, conduction disturbances, and sudden death. We evaluated the value of contrast-enhanced multi detector computed tomography in delineating myocardial scar and granulomatous inflammation by comparing our findings with gadolinium magnetic resonance in a patient diagnosed with cardiac sarcoidosis. PMID- 16257461 TI - Concomitant presence of subtypes of left coronary artery disease. AB - OBJECTIVE: To study the risk profile associated with surgical revascularization for mixed left main and left main equivalent diseases. METHODS AND RESULTS: Retrospective analysis of medical records of patients who underwent CABG from January 1997 and December 2002 were studied. 210 patients were included of which 30.5% (64) had left main, 49.5% (104) had left main equivalent and 20% (42) had mixed type of stenotic disease. The mixed type left coronary disease was associated with very high post-operative mortality. The factors associated with mortality were degree of left main artery stenosis, cardiopulmonary bypass pump time, no. of packed cell units transfused during surgery, post-operative low cardiac output, re-exploration of chest and no. of days spent as intubated. Low cardiac output, chest re-exploration and no. of days intubated were independently associated with mortality. CONCLUSIONS: "Mixed left main and left main equivalent disease" makes a large proportion of left sided CAD. It is a unique subtype associated with very high risk, and it should be treated with extra caution. Characterization of its risk profile should be done with larger studies to improve the outcome. PMID- 16257462 TI - Inflammatory markers are not associated with outcomes following elective external cardioversion. AB - BACKGROUND: Electrical cardioversion is a common modality of therapy for persistent atrial fibrillation. Unfortunately even if the cardioversion is initially successful many patients revert to atrial fibrillation. It has been proposed that there may be an inflammatory component to this arrhythmia. It is interesting to speculate that this may have a role in determining the outcome following elective cardioversion. METHODS: The study group consisted of 81 patients with persistent atrial fibrillation undergoing elective external cardioversion. Blood samples were taken immediately prior to the procedure. Soluble E-Selectin, P-Selectin, intra-cellular adhesion molecule and vascular cell adhesion molecule were assayed using a commercially available enzyme linked immunosorbent assay technique (R&D systems) and high sensitivity C reactive protein was measured by rate nephelometry. Patients were reviewed at 8 weeks and bloods were taken at this time. RESULTS: At baseline patients who had an unsuccessful cardioversion (n=15) were compared to those who had a successful cardioversion (n=66). Thirty-two patients of the 66 initially successful patients reverted to atrial fibrillation during the follow-up period. There was no difference in the levels of baseline serum inflammatory markers measured between those with an unsuccessful cardioversion and those who were successful. When the group who reverted to atrial fibrillation were compared to those who remained in sinus rhythm again there was no difference in the levels of serum markers measured at baseline. CONCLUSION: There was no association between maintenance of sinus rhythm following cardioversion and serum inflammatory markers. PMID- 16257463 TI - Takotsubo cardiomyopathy. AB - Recently, many reports have been published about apical ballooning or takotsubo cardiomyopathy. This disease is a new entity of acute reversible heart failure. Abnormal catecholamine dynamics due to stress may be the primary cause of this condition. PMID- 16257464 TI - Public health advocate. AB - With antimicrobial resistance mounting, an important public health goal is to preserve therapeutic effectiveness of remaining antimicrobials. To that end, fewer antimicrobials should be used in human medicine and in agriculture. Public health initiatives to reduce antimicrobial overuse could benefit from concurrent collection of agricultural usage data; they could aid scientific understanding of the resistance problem and confirm the efficacy of interventions. Data collection in this context should be a priority. However, usage data are nonessential to achieving the public health goal. U.S. regulation of agricultural antimicrobials today is very reliant on risk assessment. While more data can be useful for use in risk assessment, microbial risk assessment itself may not be well suited to the purpose of reducing antimicrobial overuse. Among other recognized shortcomings, current microbial risk assessment models typically fail to account for the essential ecological nature of antimicrobial resistance. This makes it inadequate for fully characterizing the human health or ecological risks of animal antimicrobials. European success at phasing out unnecessary antimicrobial usage in agriculture, on the other hand, has derived from decisions based on public health concerns and political will, and not on the collection of usage data or on the successful completion of a risk assessment. PMID- 16257465 TI - Stakeholder position paper: epidemiological perspectives on antibiotic use in animals. AB - Epidemiologists studying antimicrobial resistance are often interested in analyzing the association between antimicrobial resistance and antimicrobial use in animals, and on the impact of antimicrobial use in animals on the occurrence of resistance in bacteria affecting human populations. Given the various potential antimicrobial use data sources, it seems likely there will be some variability in the utility of the data for interpreting trends in antimicrobial resistance and investigating the relationship between antimicrobial use in animals and antimicrobial resistance in bacteria affecting human health. From an epidemiologic perspective, the major issues related to incorporation of antimicrobial use data into antimicrobial resistance monitoring programs are the further development of epidemiologic methods for collecting, quantifying, analyzing and interpreting use data; an open and realistic consideration of the limitations of the data; developing an understanding of scaling, temporal and spatial heterogeneity issues; and the interpretative problems of ecologic and atomistic fallacy. Given the many potential biases in antimicrobial use data, attempts to relate levels of antimicrobial use to levels of antimicrobial resistance should be done with caution until the data are better understood and the aforementioned issues have been addressed. PMID- 16257466 TI - Coping mechanisms, perception of health and cardiovascular dysfunction in Africans. AB - The purpose of this study was to compare active and passive coping strategies of Africans with perception of own health and cardiovascular data. The subjects included 236 apparently healthy Africans (men=109; women=127). The COPE questionnaire was adapted, translated and validated for Africans. Scores on reliable sub-scales were used to classify men and women into more active coping (AC) and more passive coping (PC) subgroups. The General Health Questionnaire measured subjective perception of health. Blood pressure was recorded before and during application of the handgrip test, using the Finapres, a continuous non invasive blood pressure monitor. Plasma renin activity (PRA) values, measured with radio immuno assay, were compared to blood pressure variables. Analyses of co-variance, adjusted for resting values and age, indicated that PC men responded with a larger increase in total peripheral resistance (TPR) (p=0.006), larger decrease in stroke volume (p=0.07), smaller increase in cardiac output (p=0.09) and larger increases in PRA resting (p=0.04) and reactivity (p< or =0.05) values. PC subjects reported a more negative perception of health than AC subjects. Young PC women presented greater hypertension prevalence rates (p< or =0.01) than AC women. In conclusion, all AC and PC subjects reacted with increased vascular reactivity on the handgrip test. PC men presented enhanced vascular reactivity, PRA and perception of poorer health values. PMID- 16257467 TI - DNA typing methods for differentiation of yeasts related to dry-cured meat products. AB - RFLP analysis of the ITS and 18S rDNA, RAPD-PCR using mini- and microsatellite primers and RFLP analysis of mitochondrial DNA were examined to discriminate yeasts related to dry-cured meat products at species and strain level. Seven species and 35 strains of yeasts usually found in dry-cured meat products were tested. RFLP analysis of the ITS1-5.8S rDNA-ITS2 and 18S rDNA did not allow the separation at species level of all of the species tested. RAPD with a M13 primer was found to be useful for differentiation of Rhodotorula mucilaginosa, Candida zeylanoides, Yarrowia lipolytica, Debaryomyces hansenii and Saccharomyces cerevisiae. However, no differences were observed between Debaryomyces polymorphus and Pichia carsonii. RAPD analysis with microsatellite primers (GACA)(4), (GTG)(5) and (GAC)(5) enabled discrimination at species and strain level. However, the degree of discrimination by means of RAPD-PCR depends highly on the primers used. Thus, the PCR fingerprinting with primer (GACA)(4) enabled a higher level of discrimination than primers (GAC)(5) and (GTG)(5). The RFLP analysis of mtDNA allowed the discrimination at the species and strain level except for R. mucilaginosa, where no polymorphisms were observed in the strains tested. RAPD analysis with primer (GACA)(4) and the restriction analysis of mtDNA used in the present work are useful for the differentiation at species and strain level of yeasts related to dry-cured meat products. PMID- 16257468 TI - On-demand release by ultrasound from osmotically swollen hydrophobic matrices. AB - Ethylene-vinyl acetate copolymer (EVAc) based controlled release systems are composed of a continuous hydrophobic polymer phase and of dispersed solid drug particles. In matrices with high drug loadings (> or =50%) most drug particles are interconnected. Thus, when these matrices are immersed in water, release rates are relatively high. In matrices with low drug loadings (<40%) most drug particles are isolated within the continuous hydrophobic polymer matrix. Thus, release rates are very low. Matrices containing water soluble particles at a low loading swell intensely after immersion in water; this is caused by the osmotic force of the isolated particles which causes the water to permeate into the hydrophobic polymer. The enclosed drug particles absorb water, and at the end of the swelling process the matrices consist of an immense number of fluid pockets containing dissolved drugs. In attempt to develop on-demand release systems, we studied the effect of ultrasound (US) on the release rates of these swollen matrices. We found that low-frequency ultrasound (20 kHz) reversibly increased the release rates from these swollen matrices by a factor of 30-500, in contrast to the unswollen matrices, where the release rates increased only by a factor of 2-3. Unswollen and swollen matrices react differently to US, as unswollen matrices contain drug particles which absorb ultrasound on the surface of the matrices, causing ultrasound attenuation. On the other hand, swollen matrices contain a compact arrangement of fluid pockets separated from each other by thin membranes containing a solution of the dissolved drug, therefore ultrasound penetrates through the matrix volume. Ultrasound penetration causes tearing of these membranes within the matrices. As a result, the fluid pockets interconnect, and drug molecules diffuse out through these interconnected pockets. PMID- 16257469 TI - Pharmacokinetics of DTPA entrapped in conventional and long-circulating liposomes of different size for plutonium decorporation. AB - The aim of the present study was to develop an efficient DTPA liposome formulation designed for plutonium decorporation. DTPA was encapsulated in conventional (CL) and polyethylene glycol-coated stealth liposomes (SL) prepared by extrusion followed by the freeze-thawing method and sizing from around 100 to 800 nm. DTPA encapsulation percentages were approximately 30% in CL of any size but dropped from 48% to 7% as the diameter of SL was reduced. The pharmacokinetics of [(14)C]-DTPA encapsulated in large and small vesicles was evaluated in rats after a single intravenous administration. Both liposomal composition and size reduction had a significant impact on pharmacokinetic parameters, inducing a marked increased in exposure of the body to DTPA and its delayed excretion. DTPA distribution was moderate in liver but enhanced in spleen and bone and was dose-dependent, especially when SL of 100 nm were given. In conclusion, small and stealth(R) vesicles have interesting properties in delivering DTPA to contaminated tissues. PMID- 16257471 TI - Co-expression of the 5-HT(3B) subunit with the 5-HT(3A) receptor reduces alcohol sensitivity. AB - Allosteric modulation of mouse 5-Hydroxytryptamine(3A) (5-HT(3A)) and 5-HT(3A/B) receptor function by ethanol and trichloroethanol (TCEt) was assessed in HEK293 cells with whole cell patch-clamp electrophysiological recordings. Ethanol enhanced 5-HT(3A) receptor function, but had no effect on mouse 5-HT(3A/B) receptor mediated currents. The enhancing action of trichloroethanol (TCEt) on mouse 5-HT(3A/B) receptor function was much less than that observed in the mouse 5-HT(3A) receptor. Where alcohol-induced increases in peak amplitude were observed, the slope of the 20-80% rising phase of current onset was also enhanced, suggesting that increases in activation rate may be one mechanism through which alcohols enhance function of the 5-HT(3) receptors. PMID- 16257472 TI - Tyrosine phosphorylation of the N-methyl-D-aspartate receptor is enhanced in synaptic membrane fractions of the adult rat hippocampus. AB - Hippocampal N-methyl-D-aspartate receptors (NMDARs) contribute to the expression of certain types of synaptic plasticity, such as long-term potentiation (LTP). It is well documented that tyrosine kinases increase NMDAR phosphorylation and potentiate NMDAR function. However, it is unclear how these phosphorylation changes result in enhanced NMDAR activity. We previously reported that NMDAR surface expression can be increased by LTP-inducing stimulation via tyrosine kinase-dependent mechanisms in the adult hippocampus [D.R. Grosshans, D.A. Clayton, S.J. Coultrap, M.D. Browning, Nat. Neurosci., 5 (2002) 27-33]. We therefore hypothesized that tyrosine phosphorylation of the NMDAR may enhance the trafficking of the receptor to the synaptic membrane. Here, we show that the stoichiometry of NR2A and NR2B tyrosine phosphorylation is significantly higher in synaptosomal membranes than intracellular microsomal/light membranes. Interestingly, NR2B tyrosine-1472, but not NR1 serine-896 or -897, phosphorylation is significantly higher in synaptosomal membranes than intracellular microsomal/light membranes. Furthermore, treatment of hippocampal slices with either a tyrosine phosphatase inhibitor or a tyrosine kinase inhibitor alters NMDAR tyrosine phosphorylation and produces a corresponding change in the concentration of NMDARs in the synaptosomal membrane fraction. Taken together, these data support the hypothesis that tyrosine phosphorylation may enhance NMDAR activity by increasing the number of NMDARs at the synaptic membrane. PMID- 16257470 TI - Causes of oncogenic chromosomal translocation. AB - Non-random chromosomal translocations are frequently associated with a variety of cancers, particularly hematologic malignancies and childhood sarcomas. In addition to their diagnostic utility, chromosomal translocations are increasingly being used in the clinic to guide therapeutic decisions. However, the mechanisms that cause these translocations remain poorly understood. Illegitimate V(D)J recombination, class switch recombination, homologous recombination, non homologous end-joining and genome fragile sites all have potential roles in the production of non-random chromosomal translocations. In addition, mutations in DNA-repair pathways have been implicated in the production of chromosomal translocations in humans, mice and yeast. Although initially surprising, the identification of these same oncogenic chromosomal translocations in peripheral blood from healthy individuals strongly suggests that the translocation is not sufficient to induce malignant transformation, and that complementary mutations are required to produce a frank malignancy. PMID- 16257473 TI - Comparison of basal and D-1 dopamine receptor agonist-stimulated neuropeptide gene expression in caudate-putamen and nucleus accumbens of ad libitum fed and food-restricted rats. AB - Behavioral studies have demonstrated that chronic food restriction augments the rewarding and motor-activating effects of centrally injected psychostimulants and direct dopamine (DA) receptor agonists. Recently, it has been shown that intracerebroventricular (i.c.v.) injection of the D-1 DA receptor agonist, SKF 82958, produces an enhanced locomotor-activating effect as well as increased activation of striatal ERK 1/2 MAP kinase, CaM kinase II, CREB, and c-fos in food restricted (FR) relative to ad libitum fed (AL) rats. Striatal neurons that express the D-1 DA receptor coexpress dynorphin and substance P, and CREB is known to couple D-1 DA receptor stimulation to preprodynorphin (ppD) gene expression. The purpose of the present study was to examine possible genomic consequences of FR using real-time quantitative RT-PCR to measure striatal neuropeptide gene expression 3 h after i.c.v. injection of SKF-82958 (20 microg). Results indicate that, in nucleus accumbens (NAc), basal levels of ppD and preprotachykinin (ppT) mRNA are lower in FR than AL rats. This may reflect a decrease in tonic DA transmission during FR which precedes the compensatory upregulation of postsynaptic D-1 DA receptor-mediated cell signaling. In response to SKF-82958 challenge, however, FR subjects displayed greater levels of ppD and ppT mRNA in NAc than did AL subjects. A similar trend was seen in caudate-putamen (CPu). SKF-82958 also increased preproenkephalin (ppE) mRNA in Nac, but not CPu, with no difference between feeding groups. The present findings regarding ppD and ppT are consistent with prior findings of increased behavioral and cellular responses to acute D-1 DA agonist challenge in FR rats. The functional consequences of increased neuropeptide gene expression in response to acute drug challenge remain to be investigated but may include modulation of behavioral effects that emerge with repeated drug exposure, including sensitization, tolerance, and addiction. PMID- 16257474 TI - Food reinforcement. AB - The reinforcing value of food, measured by how hard someone is willing to work to obtain food, is influenced by food palatability, food deprivation and food variety, and may be a more powerful determinant of food intake than hedonics or liking. The reinforcing value of food is mediated in part by dopaminergic activity. Genotypes that influence dopamine transport and the density of dopamine D2 receptors interact with food reinforcement to influence eating behavior, and D2 receptor genotypes may influence food reinforcement and weight gain after smoking cessation. Inhibition of dopamine transport increases brain dopamine concentrations, which may influence weight gain after smoking cessation and can reduce energy intake in obese adults. PMID- 16257475 TI - The mouse cerebellum from 1 to 34 months: parallel fibers. AB - Synaptic losses are critical events in the aging brain. In the cerebellum, synapses between the parallel fibers and Purkinje cells are strategic elements of its neuronal circuitry. We have examined the number of synaptic varicosities per parallel fiber in the anterior lobe of the cerebellum in young (1, 2.5 and 5 months), mature (10 and 18 months), and old (26 and 34 months) C57BL/6 mice. This number peaks at 2.5 months and starts to decrease immediately. This decrease is due to a reduction of the linear density of synaptic varicosities along the parallel fibers and a shortening of the parallel fibers. The fastest and most dramatic decline occurs during young adulthood between 2.5 and 5 months when 59% of the synaptic varicosities are lost. Accelerated loss also occurs in old age between 26 and 34 months. Changes between 5 and 26 months are the slowest. Between 2.5 and 34 months, mice lose 87% of their parallel fiber synaptic varicosities. Unlike many post-reproductive processes of aging, the early-onset decline beginning at 2.5 months may be under strong natural selection pressure. PMID- 16257476 TI - High-fat diets, insulin resistance and declining cognitive function. AB - Results from our work in rats and others findings from human epidemiologic studies demonstrate deficits in cognitive performance following chronic ingestion of high fat, high saturated fat, diets. Yet, the precise physiologic mechanism underlying these deficits is not well understood. We report that older adults with insulin resistance show remarkably similar deficits in cognitive function and respond to glucose ingestion in a comparable manner to rodents fed a high-fat diet, suggesting that insulin resistance is a probable mediator of these diet induced deficits. As insulin resistance worsens to overt type 2 diabetes, profound deficits in cognitive functions, especially those dependent on the medial temporal lobes, are apparent in both obese Zucker rats and humans with type 2 diabetes. Unlike the older adult with insulin resistance, glucose ingestion further impairs medial temporal lobe function in adults with type 2 diabetes. Collectively, the human and rodent data point to a role of diet-induced endocrine abnormalities, including the development of insulin resistance, as mediating the cognitive deficits associated with high fat consumption. PMID- 16257477 TI - Social and biological determinants of cognitive aging. AB - The aging of populations increases the importance of cognitive function as a public health issue. The Whitehall II study is investigating influences on aging processes within social context, with tests in five domains of cognitive function (short-term memory, inductive reasoning, vocabulary, phonemic and semantic fluency). The tests have now been completed three times over a 10-year period, by participants initially aged 40-60 years. In mid-life, socioeconomic differences in cognitive function were large, while APOE genotype had little influence. Higher occupational status was strongly related to lower metabolic syndrome prevalence. This social-biological gradient illustrates that potential confounding of psychosocial, behavioral and biological effects on cognitive decline is an important consideration in etiologic research. Additionally, it highlights the importance for population health of the 'causes of the causes' of cognitive aging. Epidemiology contributes to our understanding of risk and protective factors for cognitive decline by showing the links between the wider determinants of health and biological markers such as glucose intolerance and the metabolic syndrome. PMID- 16257479 TI - Fowlpox virus vaccines for HIV and SHIV clinical and pre-clinical trials. AB - DNA prime and recombinant fowlpox virus (rFPV) boost vaccines were designed to express multiple HIV or SIV antigens for use in human clinical trials and in pre clinical trials in macaques. Three sets of vaccines with matching HIV or SIV antigen sets, modified for vaccine safety considerations, were constructed and shown to express the relevant proteins. The rFPV vaccines with inserts at up to three sites, were stable on passage in chick cell culture, including during GMP manufacture of vaccines for human Phase I clinical trials. Cellular and humoral immunogenicity in mice was demonstrated using a DNA prime/rFPV boost and vaccinia virus challenge model. These data establish a preliminary safety and efficacy profile for these multigenic vaccines suggesting they are suitable for advanced development as candidate HIV vaccines. PMID- 16257478 TI - Chaperonin GroEL and its mutant D87K protect from ischemia in vivo and in vitro. AB - Protein aggregation and misfolding are central mechanisms of both acute and chronic neurodegeneration. Overexpression of chaperone Hsp70 protects from stroke in animal and cell culture models. Although it is accepted that chaperones protect cells, the mechanism of protection by chaperones in ischemic injury is poorly understood. In particular, the relative importance of preventing protein aggregation compared to facilitating correct protein folding during ischemia and recovery is not known. To test the importance of protein folding and minimize interaction with co-chaperones we studied the bacterial chaperonin GroEL (HSPD1) and a folding-deficient mutant D87K. Both molecules protected cells from ischemia like injury, and reduced infarct volume and improved neurological outcome after middle cerebral artery occlusion in rats. Protection was associated with reduced protein aggregation, assessed by ubiquitin immunohistochemistry. Marked neuroprotection by the folding-deficient chaperonin demonstrates that inhibition of aggregation is sufficient to protect the brain from ischemia. This suggests that strategies to maintain protein solubility and inhibit aggregation in the face of acute insults such as stroke may be a useful protective strategy. PMID- 16257480 TI - Process development for the mass production of Ehrlichia ruminantium. AB - This work describes the optimization of a cost-effective process for the production of an inactivated bacterial vaccine against heartwater and the first attempt to produce the causative agent of this disease, the rickettsia Ehrlichia ruminantium (ER), using stirred tanks. In vitro, it is possible to produce ER using cultures of ruminant endothelial cells. Herein, mass production of these cells was optimized for stirring conditions. The effect of inoculum size, microcarrier type, concentration of serum at inoculation time and agitation rate upon maximum cell concentration were evaluated. Several strategies for the scale up of cell inoculum were also tested. Afterwards, using the optimized parameters for cell growth, ER production in stirred tanks was validated for two ER strains (Gardel and Welgevonden). Critical parameters related with the infection strategy such as serum concentration at infection time, multiplicity and time of infection, and medium refeed strategy were analyzed. The results indicate that it is possible to produce ER in stirred tank bioreactors, under serum-free culture conditions, reaching a 6.5-fold increase in ER production yields. The suitability of this process was validated up to a 2-l scale and a preliminary cost estimation has shown that the stirred tanks are the least expensive culture method. Overall, these results are crucial to define a scaleable and fully controlled process for the production of a heartwater vaccine and open "new avenues" for the production of vaccines against other ehrlichial species, with emerging impact in human and animal health. PMID- 16257481 TI - X-ray diagnosis of acute scaphoid fractures. AB - In a retrospective review of the radiographs taken for 113 acute scaphoid fractures, each view was assessed for the clarity of demonstration of the fracture. The X-rays on which diagnosis of fracture were made, were taken between 0 and 16 days after injury (mean, 2 days). Whenever a lateral, supinated oblique or elongated view was taken, the fracture was always seen clearly on an alternative view. We recommend the use of four views at initial presentation of suspected scaphoid fracture: PA and lateral to assess carpal alignment, with pronated oblique and ulnar deviated PA to detect the fracture. PMID- 16257482 TI - Ozone sensitivity of currant tomato (Lycopersicon pimpinellifolium), a potential bioindicator species. AB - The wild tomato species Lycopersicon pimpinellifolium (currant tomato) was exposed to different O3 concentration, both in controlled environment fumigation facilities and in open-top chambers, to assess its sensitivity and to verify its potential as a bioindicator plant. Plants appeared particularly sensitive to O3 at an early stage of growth, responding with typical chlorotic spots within 24 h after exposure to a single pulse of 50 ppb for 3 h, and differentiating peculiar symptoms, such as reddish necrotic stipples, bronzing and extensive necrosis, depending on O3 concentration. Histo-cytochemical investigations with 3,3' diaminobenzidine, to localize H2O2, and Evans blue, to detect dead cells, suggested that currant tomato sensitivity to O3 could be due to a deficiency in the anti-oxidant pools. The combination of these stainings proved to be useful, either to predict visible symptoms, early before their appearance, and to validate leaf ozone injury. PMID- 16257483 TI - Ovarian metastases from an urachal adenocarcinoma. PMID- 16257484 TI - The promoter of the human sodium/iodide symporter responds to certain phthalate plasticisers. AB - The diesters of benzene-1,2-dicarboxylic (phthalic) acid, the phthalates, are used to make plastics flexible and can comprise 40% of the weight of plastic. Human exposure to phthalates can occur via ingestion, inhalation and dermal routes, as well as through parenteral exposure from medical devices containing phthalates. Since earlier morphological studies showed that some phthalates induced thyroid hyperactivity, we thought it important to investigate possible effects of six major phthalates on the transcriptional activity of sodium/iodide symporter (NIS). Di-isodecyl phthalate (DIDP), benzyl butyl phthalate (BBP) and di-octyl phthalate (DOP) increased the activity of the human NIS promoter construct 2.5-, 2.6- and 2.4-fold, respectively. Likewise, these phthalates also enhanced the rat NIS endogenous mRNA expression ca. 2-fold. No effect was observed for bis-(2-ethylhexyl) phthalate (DEHP) and di-isononyl phthalate (DINP), whereas dibutyl phthalate (DBP) appeared to down-regulate hNIS promoter. Although the demonstrated stimulation of NIS gene transcription by DIDP, BBP and DOP is not very strong, this finding is of great importance as humans are routinely exposed for long periods to phthalate plasticisers, the accumulation of which may contribute to thyroid hyperfunction. PMID- 16257486 TI - Chemosensory anxiety signals augment the startle reflex in humans. AB - The aim of the present study was to investigate whether chemosensory anxiety signals can activate behavioral withdrawal systems in humans. Twelve male university students donated their axillary sweat in two situations: right before an oral academic examination (anxiety condition) and during ergometric training (exercise condition). Subjective ratings revealed that the odor donors experienced significantly more anxiety and less pleasure during the anxiety condition than during the exercise condition. Seven subjects (three females) participated in the psychophysiological experiment. The chemosensory stimuli from pooled sweat samples of the donors, and from unused cotton pads (pad condition) were presented via a constant-flow olfactometer. Acoustic startle probes (100 dB (A)) were delivered during and between the presentations of the chemosensory stimuli. Only three subjects were able to discriminate the chemosensory stimuli of the human sweat samples from room air. However, the startle reflex amplitude (EMG of the eyeblink response) recorded in the context of chemosensory anxiety signals was increased, as compared to the amplitude recorded in the context of chemosensory stimuli from either exercise (p = 0.018) or cotton pad (p = 0.012). It is concluded that chemosensory anxiety signals may pre-attentively prime defensive behavior. PMID- 16257485 TI - X-ray absorption spectroscopy study of a copper-containing material after thermal treatment. AB - Thermal immobilization of copper contaminant in a copper-containing solid material collected from local copper smelting and foundry area is investigated in the present work. X-ray absorption spectroscopy (XAS) and X-ray diffraction (XRD) are employed for copper speciation. XAS results indicate that cupric hydroxide is the major copper species in the solid material dried at 105 degrees C. After being subjected to a 500 degrees C thermal process, cupric hydroxide still remains as the main copper species, but some Cu(II) is chemically reduced to Cu(I). More cupric hydroxide is progressively converted to Cu(I) as the sample was heated at 1100 degrees C than that heated at 500 degrees C. The sample heated at 500 degrees C is in its original powder form. However, thermal treatment at 1100 degrees C transforms the powder into a hardened granule-like form that is much bigger in size and difficult to be ground into powders. The sample is sintered with the sparingly soluble cuprous oxide and elemental copper being encapsulated inside. Toxicity characteristic leaching procedure (TCLP) results depict that amount of copper leached from the sample (containing 133,000 mg copper kg-1) heated at 1100 degrees C for 2 h is considerably minor, being 367 mg copper kg-1. PMID- 16257487 TI - The effect of changed visual feedback on intention tremor in multiple sclerosis. AB - In patients with multiple sclerosis (MS), intention tremor amplitude is enhanced during the visually guided compared to the memory guided motor tasks. In the present study, the effect of visual feedback on intention tremor was investigated during visually guided wrist step-tracking tasks. Specifically, visual feedback of the hand was provided either instantly or averaged over different time windows. Thirteen MS patients with intention tremor and 14 healthy controls performed the wrist step-tracking task, while the visual representation of the actual hand position was displayed instantly or averaged over time windows of 150, 250 and 350 ms. It has been found in the patient group that, in association with a decreased initial error and decreased tremor amplitude on the screen, the amplitude of the actual performed tremor also decreased when visual feedback was changed. The tremor reduction was not different between conditions with manipulated feedback, although delays in presenting visual feedback of the hand position increased when the time window was larger. The reduction in overall tremor amplitude was unlikely related to other factors, such as eye fixation deficits or the speed of the primary hand movement. These results suggest that hand tremor severity is dependent on the visual feedback of position and movement errors. PMID- 16257488 TI - Noxious heat induces fMRI activation in two anatomically distinct clusters within the nucleus accumbens. AB - Using functional magnetic resonance imaging (fMRI) we found that a noxious thermal stimulus (46 degrees C) to the hand activates the nucleus accumbens (NAc) in humans, while a non-noxious warm stimulus (41 degrees C) does not. Following the noxious stimulus, two distinct foci of decreased activation were observed showing distinct time course profiles. One focus was anterior, superior, and lateral and the second that was more posterior, inferior, and medial. The anatomical segregation may correlate with the functional components of the NAc, i.e., shell and core. The results support heterogeneity of function within the NAc and have implications for the understanding the contribution of NAc function to processing of pain and analgesia. PMID- 16257489 TI - The regulation of rotenone-induced inflammatory factor production by ATP sensitive potassium channel expressed in BV-2 cells. AB - Our previous studies have demonstrated that activating ATP-sensitive potassium channel (K(ATP) channel), not only improved Parkinsonian behavior and neurochemical symptoms, but also reduced iNOS activity and mRNA levels in striatum and nigra of rotenone rat model of Parkinson's disease (PD). In this study, it was first shown that the subunits of K(ATP) channels are expressed in BV-2 cells, and then it was investigated whether K(ATP) channel was involved in regulating inflammatory factor production from BV-2 cells activated by rotenone. It was found that K(ATP) channel was expressed in BV-2 cell and formed by the combination of Kir 6.1 and SUR 2A/2B. K(ATP) channel openers (KCOs) including pinacidil, diazoxide and iptakalim (Ipt) exerted beneficial effects on rotenone induced morphological alterations of BV-2 cells, decreased tumor necrosis factor alpha (TNF-alpha) production and the expression and activity of inducible isoform of nitric oxide synthase (iNOS). Either glibenclamide or 5-hydroxydecanoate acid (a selective mitochondrial K(ATP) channel blocker) could abolish the effects of KCOs, suggesting that K(ATP) channels, especially mitochondrial ATP-sensitive potassium channels (mitoK(ATP) channels), played a crucial role in preventing the activation of BV-2 cells, and subsequently the production of a variety of proinflammatory factors. Therefore, activation of K(ATP) channel might be a new therapeutic strategy for treating neuroinflammatory and neurodegenerative disorders. PMID- 16257490 TI - Neonatal manipulation of oxytocin affects expression of estrogen receptor alpha. AB - In adult females many of the effects of the neuropeptide oxytocin are steroid, and especially estrogen dependent. Here we demonstrate for the first time that neonatal manipulation of oxytocin can affect the expression of estrogen receptor alpha. On the first day of postnatal life male and female prairie voles (Microtus ochrogaster) were randomly assigned to receive one of four treatments; (a) 50 microl i.p. injection of 3 microg oxytocin (approximately 1 microg/g), (b) 0.3 microg of an oxytocin antagonist (approximately 0.1 microg/g), or (c) isotonic saline. A fourth group was handled, but not injected. On postnatal day 8 or 21, brain tissue was collected, fixed and sectioned. Free-floating sections were stained for estrogen receptor alpha using immunocytochemistry, and estrogen receptor alpha immunoreactive neurons were compared by age, treatment, and sex. To compare the temporal expression of estrogen receptor alpha an additional set of brains was collected from untreated males and females on the day of birth. The effects of oxytocin manipulations were age dependent, sexually dimorphic, and site-specific. While there were no significant treatment effects on postnatal day 8, by postnatal day 21 females that received oxytocin showed a significant increase in the number of cells expressing estrogen receptor alpha immunoreactivity in the ventromedial nucleus of the hypothalamus. Treatment with oxytocin antagonist resulted in a significant decrease in estrogen receptor alpha immunoreactivity in the medial preoptic area in postnatal day 21 females. While there were no significant effects in males, males treated with oxytocin antagonist trended toward a reduction in estrogen receptor alpha-immunoreactivity in the medial amygdala. The results indicate that oxytocin can have organizational effects on the expression of estrogen receptor alpha, that these effects are sexually dimorphic, and finally that during the preweaning period the development of estrogen receptor alpha is sexually dimorphic. PMID- 16257491 TI - Gene expression in the rat brain during sleep deprivation and recovery sleep: an Affymetrix GeneChip study. AB - Previous studies have demonstrated that macromolecular synthesis in the brain is modulated in association with the occurrence of sleep and wakefulness. Similarly, the spectral composition of electroencephalographic activity that occurs during sleep is dependent on the duration of prior wakefulness. Since this homeostatic relationship between wake and sleep is highly conserved across mammalian species, genes that are truly involved in the electroencephalographic response to sleep deprivation might be expected to be conserved across mammalian species. Therefore, in the rat cerebral cortex, we have studied the effects of sleep deprivation on the expression of immediate early gene and heat shock protein mRNAs previously shown to be upregulated in the mouse brain in sleep deprivation and in recovery sleep after sleep deprivation. We find that the molecular response to sleep deprivation and recovery sleep in the brain is highly conserved between these two mammalian species, at least in terms of expression of immediate early gene and heat shock protein family members. Using Affymetrix Neurobiology U34 GeneChips , we also screened the rat cerebral cortex, basal forebrain, and hypothalamus for other genes whose expression may be modulated by sleep deprivation or recovery sleep. We find that the response of the basal forebrain to sleep deprivation is more similar to that of the cerebral cortex than to the hypothalamus. Together, these results suggest that sleep-dependent changes in gene expression in the cerebral cortex are similar across rodent species and therefore may underlie sleep history-dependent changes in sleep electroencephalographic activity. PMID- 16257492 TI - Hyperactive striatal neurons in symptomatic Huntington R6/2 mice: variations with behavioral state and repeated ascorbate treatment. AB - Membrane and morphological abnormalities occur in the striatum of R6/2 transgenics, a widely used mouse model of Huntington's disease. To assess changes in behavior-related neuronal activity, we implanted micro-wire bundles in the striatum of symptomatic R6/2 mice and wild-type controls. Unit activity was recorded in an open-field arena once weekly for the next several weeks. For each recording session, firing rate was monitored before, during, and after a period of light anesthesia to assess the influence of behavioral arousal. Because low ascorbate in striatal extracellular fluid may contribute to Huntington's disease symptoms, all animals received an injection of either 300 mg/kg sodium ascorbate or vehicle for three consecutive days prior to each recording session. In R6/2 mice, regardless of treatment, striatal unit activity was significantly faster than in wild-type controls. The difference in mean (+/-S.E.M.) firing was most apparent during wakefulness (6.4+/-0.8 vs. 3.5+/-0.3 spikes/s) but also persisted during anesthesia (2.0+/-0.3 vs. 0.7+/-0.1 spikes/s). Assessment of treatment duration indicated that R6/2 mean waking discharge rate was significantly slower after three weeks than after one week of ascorbate treatment (3.1+/-0.6 vs. 10.2+/-2.7 spikes/s). Vehicle-treated R6/2s showed no such decline in striatal activity ruling out an age- or injection-related effect. Slow-scan voltammetry in separate animals confirmed that ascorbate-injections returned the level of striatal extracellular ascorbate in R6/2 mice to that of wild-type controls. Our results indicate that although striatal neurons modulate firing in relation to behavioral state, impulse activity is consistently elevated in transgenic relative to wild-type mice. Restoring extracellular ascorbate to the wild-type level reverses this effect suggesting a role for ascorbate in normalizing neuronal function in Huntington's disease striatum. PMID- 16257493 TI - Differential effect of alpha-syntrophin knockout on aquaporin-4 and Kir4.1 expression in retinal macroglial cells in mice. AB - Aquaporin-4 water channels and the inwardly rectifying potassium channels Kir4.1 are coexpressed in a highly polarized manner at the perivascular and subvitreal endfeet of retinal Muller cells and astrocytes. The present study was aimed at resolving the anchoring mechanisms responsible for the coexpression of these molecules. Both aquaporin-4 and Kir4.1 contain PDZ-domain binding motifs at their C-termini and it was recently shown that mice with targeted disruption of the dystrophin gene display altered distribution of aquaporin-4 and Kir4.1 in the retina. To test our hypothesis that alpha-syntrophin (a PDZ-domain containing protein of the dystrophin associated protein complex) is involved in aquaporin-4 and Kir4.1 anchoring in retinal cells, we studied the expression pattern of these molecules in alpha-syntrophin null mice. Judged by quantitative immunogold cytochemistry, deletion of the alpha-syntrophin gene causes a partial loss (by 70%) of aquaporin-4 labeling at astrocyte and Muller cell endfeet but no decrease in Kir4.1 labeling at these sites. These findings suggest that alpha-syntrophin is not involved in the anchoring of Kir4.1 and only partly responsible for the anchoring of aquaporin-4 in retinal endfeet membranes. Furthermore we show that wild type and alpha-syntrophin null mice exhibit strong beta1 syntrophin labeling at perivascular and subvitreal Muller cell endfeet, raising the possibility that beta1 syntrophin might be involved in the anchoring of Kir4.1 and the alpha syntrophin independent pool of aquaporin-4. PMID- 16257494 TI - Subtype-dependence of N-methyl-D-aspartate receptor modulation by pregnenolone sulfate. AB - N-methyl-D-aspartate receptors play a critical role in synaptogenesis, synaptic plasticity, and excitotoxicity. They are heteromeric complexes of NR1 combined with NR2A-D and/or NR3A-B subunits. The subunit composition determines the biophysical and pharmacological properties of the N-methyl-D-aspartate receptor channel complex. In this study, we report that responses mediated by recombinant rat N-methyl-D-aspartate receptors expressed in human embryonic kidney HEK293 cells are differentially affected by naturally occurring neurosteroid pregnenolone sulfate. We show that responses induced by 1mM glutamate in NR1 1a/NR2A and NR1-1a/NR2B receptors are potentiated five- to eight-fold more by pregnenolone sulfate than responses of NR1-1a/NR2C and NR1-1a/NR2D receptors with no differences in the concentration of pregnenolone sulfate that produced 50% potentiation. In addition to potentiation, pregnenolone sulfate also has an inhibitory effect at recombinant N-methyl-D-aspartate receptors, with values of the concentration of pregnenolone sulfate that produces 50% inhibition of NR1/NR2D=NR1/NR2C 10 over an 80-300 kJ mol(-1) range of internal energies. The driving force for the dissociations of 3 is provided by large Franck-Condon effects on vertical neutralization and possibly from involvement of excited electronic states. Calculations also provided the adiabatic ionization energy of 3, IE(adiab) = 5.48 eV and vertical recombination energy of cation 3(+), RE(vert) = 4.70 eV. The present results strongly indicate that oxazolone structures can explain fragmentations of b-type peptide ions upon electron capture, contrary to previous speculations. PMID- 16257533 TI - Delta(DeltaS) and Delta(DeltaS) for the competing bond cleavage reactions in (CH3CN)(ROH)H+ [R = CH3, C2H5, C3H7, (CH3)2CH]. AB - Microcanonical variational transition-state theory was used to determine the entropies of activation for hydrogen-bond cleavage reactions leading to CH(3)CN + ROH(2)(+) in a series of acetonitrile-alcohol proton-bound pairs (CH(3)CN)(ROH)H(+) (where R = CH(3), CH(3)CH(2), CH(3)CH(2)CH(2), and (CH(3))(2)CH). In each case, the dissociation potential surface was modelled at the MP2/6-31 + G(d) level of theory. The dissociating configurations having the minimum sums-of-states were identified in each case and the resulting entropies of activation were calculated. Combined with previous work on the competing reaction leading to CH(3)CNH(+) + ROH, the results permitted the determination of the Delta(DeltaS) in each proton-bound pair. For the (CH(3)CN)(CH(3)OH)H(+) and (CH(3)CN)(CH(3)CH(2)OH)H(+) proton-bound pairs, the entropies of activation for the two dissociating channels are essentially the same [i.e., Delta(DeltaS) = 0], while Delta(DeltaS) for the propanol-containing pairs ranged between 40 and 45 J K(-1) mol(-1). The latter non-zero values are due to a combination of the location of the dividing surface in each dissociation and the rapidity with the frequencies of the vanishing vibrational modes go to zero as they are converted to product translations and rotations during the dissociation. PMID- 16257535 TI - Eliminating bias in the estimation of the geometric mean of HI titres. AB - The standard test for anti-haemagglutinin antibody titration is the haemagglutination inhibition (HI) test. The HI titre is defined as the dilution factor of the highest dilution that still completely inhibits haemagglutination. If the highest dilution tested (1:2560) still completely inhibits haemagglutination, an HI titre value of 2560 is assigned. Logarithmically transformed HI titres tend to be normally distributed. But because dilutions less than 1:2560 are not tested, the distribution may be truncated and the assumption of normality may not hold. As a consequence, the geometric mean titre (GMT) will be underestimated. Using data from 10 clinical studies, it is shown here that the GMT may be underestimated by 5-13%. An unbiased estimate of the GMT can be obtained by a statistical method that originates from the analysis of survival data: maximum likelihood estimation for censored observations. The maximum likelihood estimate of the GMT of truncated HI titres can be readily obtained using the statistical software package SAS. PMID- 16257534 TI - Novel transcription factor zfh-5 is negatively regulated by its own antisense RNA in mouse brain. AB - Here, we report features of a novel transcription factor zfh-5, which we isolated from the mouse brain; in addition to the mRNA, the antisense strand of zfh-5 is also expressed in the developing brain, in a manner complementary to the expression of zfh-5 mRNA. Although most neurons express zfh-5 mRNA soon after their final mitosis, several types of neurons, such as the pyramidal and granule cells in the hippocampus, express the zfh-5 antisense RNA prior to the mRNA expression. Using gene-targeting approach, we showed that this antisense RNA has a negative regulatory role on the expression of zfh-5 mRNA. These observations suggest that, in specific types of neurons, the expression of zfh-5 is additionally regulated by a mechanism depending on this antisense RNA. PMID- 16257536 TI - Feline immunodeficiency virus vaccine: implications for diagnostic testing and disease management. AB - Feline immunodeficiency virus (FIV) is a common feline pathogen, with an overall infection prevalence of approximately 11% in cats worldwide. Most infected cats eventually succumb due to direct viral effects or, more commonly, to secondary infections resulting from virus-induced immunosuppression. FIV infection is considered lifelong, and diagnosis most often relies on detection of virus specific antibodies. A currently available whole virus, adjuvanted, inactivated FIV vaccine induces antibodies in vaccinates that is indistinguishable from those induced by infection. As a result, currently available diagnostic tests cannot reliably distinguish vaccinated cats from infected cats, or from cats that are both vaccinated and infected. From both an epidemiologic and an individual cat perspective, it is impossible to determine whether use of this vaccination is more beneficial than it is harmful. PMID- 16257537 TI - Feline immunodeficiency virus vaccine: a rational paradigm for clinical decision making. AB - A veterinarian must take into consideration his/her responsibility to prevent disease when assessing the needs of a client's cat that is risk for FIV infection based on its established lifestyle. Cats infected with FIV have debilitated immune functions and exhibit a high level of chronic morbidity impacting on the animal's welfare and the owner's economic abilities to maintain the pet. Attempts to reduce the prevalence of FIV solely by advising clients to maintain their cats indoors has resulted in poor compliance and not impacted on a change in infection rates with outdoor cats. Therapeutics have not impacted on outcomes in infected animals. There has a need for a vaccine for high-risk cats. Options for vaccines that do not confound the current FIV antibody test have not been efficacious against a broad spectrum of isolates. Fel-O-Vax FIV, a conventional non-marker whole virus, has shown good efficacy against heterologous challenges. The intervention should be discussed with cat owners since the vaccine has a reasonable expectation of preventing FIV infection in cats at risk without undue safety issues. Veterinarians who do not initiate this dialogue with owners who have outdoor cats in an environment where 2.5% of cats in the USA are infected may be remiss in their professional responsibilities. PMID- 16257538 TI - Regulatory considerations for marker vaccines and diagnostic tests in the U.S. AB - Marker vaccines and diagnostic tests can prove to be invaluable in disease eradication and control programs, as was found in the pseudorabies (Aujeszky's Disease) virus eradication program in the U.S. During that campaign, numerous gene-deleted vaccines and companion diagnostic test kits were used to differentiate infected animals from vaccinated animals, in a strategy that ultimately led to eradication of the disease in commercial swine herds. The United States Department of Agriculture played a key role in delivery of that success by developing biologics policy, evaluating each product, and ensuring that the conditions of licensure were met. What was most critical in the overall eradication effort, however, was the detailed and dedicated interaction among key players: the biologics regulators, manufacturers, Federal, State, and local regulatory partners, veterinary researchers, industry associations, and animal owners. A good disease control program has to include all of these. The regulatory requirements for licensure of marker vaccines and diagnostic test kits are not different from that for other products. There are several mechanisms for vaccine approval, some more rapid than others, but only a few that could apply to these products. Generally, the platforms that might support marker vaccines and companion diagnostic kits are those based on genetic engineering or protein manipulation. If the product is derived from the application of biotechnology, then additional regulatory considerations are applicable. Most important of these are the considerations found in the National Environmental Policy Act (NEPA), wherein deliberate release of any organism containing recombinant DNA into the environment is subject to review and approval by appropriate federal agencies. Environmental release and NEPA compliance are discussed. PMID- 16257539 TI - Non-capsid proteins to identify foot-and-mouth disease viral circulation in cattle irrespective of vaccination. AB - The ability of foot-and-mouth disease virus (FMDV) to establish subclinical and even persistent infection, the so called carrier state, imposes the need to reliably demonstrate absence of viral circulation, to monitor the progress of control measures, either during eradication programs or after reintroduction of virus in free areas. This demonstration becomes critical in immunized populations, because of the concern that silent viral circulation could be hidden by immunization. This concern originates from the fact that vaccination against foot-and-mouth disease (FMD) protects against clinical disease, but not necessarily against subclinical infection or establishment of the carrier state in cattle. A novel approach, developed and validated at PANAFTOSA during the 1990s, based on an immunoenzymatic system for detection of antibodies against non capsid proteins (NCP) has proven valuable for monitoring viral circulation within and between herds, irrespective of the vaccination status. Antibodies against NCP are induced during infection but, in principle, not upon vaccination. The validation of this system led to its international recognition as the OIE index test. The fitness of this serosurvey tool to assess viral circulation in systematically vaccinated populations was demonstrated through its extensive application in most regions in South America. The experience attained in these regions supported the incorporation of the "free of FMD with vaccination" provisions into the OIE code. Likewise, it opened the way to alternatives to the "stamping out" policy. The results gave input to an old controversy related to the real epidemiological significance, if any, of carrier animals under the vaccination conditions in South America, and supported the development of recommendations and guidelines that are being implemented for serosurveys that go with control measures in vaccinated populations. PMID- 16257541 TI - Efficient detection of PrPSc (263K) in human plasma. AB - The potential contamination of human blood or plasma with prions, such as variant Creuftzfelt-Jacob disease (vCJD), is becoming a serious problem. In this study, we established a Western blot-based detection method for PrP(Sc) (263K) spiked in plasma. Although plasma contains a large amount of protein, specific detection of a small amount of 263K in plasma could be specifically detected only after ultra centrifugation followed by heat-denaturation of plasma proteins included in the resulting precipitate, before the digestion with proteinase K. PMID- 16257542 TI - Replacement of primary chicken embryonic fibroblasts (CEF) by the DF-1 cell line for detection of avian leucosis viruses. AB - International regulations prescribe that the absence of avian leucosis viruses (ALV) in avian live virus vaccines has to be demonstrated. Primary chicken embryo fibroblasts (CEF) from special SPF chicken lines are normally used for detection of ALV. The suitability of the DF-1 cell line for ALV-detection, as alternative for primary CEF, was studied in three types of experiments: (1) in titration experiments without cell passage, (2) in experiments with passages in cell cultures according to European Pharmacopoeia requirements, and (3) in experiments with commercial live avian vaccines that had been spiked with known amounts of ALV. In all tests the sensitivity of ALV-A and ALV-J detections on DF-1 cells was at least as high as on primary CEF. The sensitivity of ALV-B detection was always superior when DF-1 cells were used. ALV were detected earlier in all comparative tests when DF-1 cells were used. ALV-A, ALV-B and ALV-J all induced CPE on DF-1 cells, whereas no clear CPE was seen on CEF-cells. For reasons of sensitivity, standardisation as well as reduction of animal use, the data support the use of DF-1 cells to monitor absence of ALV in vaccine virus seed lots or finished products. PMID- 16257543 TI - Overview of avian influenza DIVA test strategies. AB - The use of vaccination in poultry to control avian influenza has been increasing in recent years. Vaccination has been primarily with killed whole virus adjuvanted vaccines. Proper vaccination can reduce or prevent clinical signs, reduce virus shedding in infected birds, and increase the resistance to infection. Historically, one limitation of the killed vaccines is that vaccinated birds cannot be differentiated serologically from naturally infected birds using the commonly available diagnostic tests. Therefore, surveillance for avian influenza becomes much more difficult and often results in trade restrictions because of the inability to differentiate infected from vaccinated animals (DIVA). Several different DIVA strategies have been proposed for avian influenza to overcome this limitation. The most common is the use of unvaccinated sentinels. A second approach is the use of subunit vaccines targeted to the hemagglutinin protein that allows serologic surveillance to the internal proteins. A third strategy is to vaccinate with a homologous hemagglutinin to the circulating field strain, but a heterologous neuraminidase subtype. Serologic surveillance can then be performed for the homologous NA subtype as evidence of natural infection. The fourth strategy is to measure the serologic response to the nonstructural protein 1 (NS1). The NS1 protein is produced in large quantities in infected cells, but it is not packaged in the virion. Since killed vaccines for influenza are primarily made with whole virions, a differential antibody response can be seen between naturally infected and vaccinated animals. However, poultry vaccines are not highly purified, and they contain small amounts of the NS1 protein. Although vaccinated chickens will produce low levels of antibody to the NS1 protein, virus infected chickens will produce higher levels of NS1 antibody, and the two groups can be differentiated. All four DIVA strategies have advantages and disadvantages, and further testing is needed to identify the best strategy to make vaccination a more viable option for avian influenza. PMID- 16257544 TI - IABs scientific workshop on neurovirulence tests for live virus vaccines, January 31-February 1, 2005, Geneva. PMID- 16257545 TI - Overview of marker vaccine and differential diagnostic test technology. AB - Recent advances in molecular biology, immunology, microbiology, genetics and microbial pathogenesis have lead to the development of a wide variety of new approaches for developing safer and more effective vaccines based on designs such as subunit vaccines, gene deleted vaccines, live vectored vaccines, and DNA mediated vaccines. Technology tools can be as basic as identifying naturally occurring strains with deletions that support differentiating infected from vaccinated animal (DIVA) needs or be based on higher technology developments such as improved protein expression and purification methods, transgenic plant- and plant virus-based antigen production, and novel adjuvants that target specific immune responses. These new approaches, when applied to the development of marker vaccines and companion diagnostic test kits hold tremendous potential for developing improved tools for eradication and control programs. Marker vaccines and companion diagnostic test kits must meet the established licensing requirements for purity, potency, safety and efficacy. Efficacy claims are based on evaluation of the level of protection demonstrated in host animal trials and may range from "prevents infection with (a specific agent)", to "for use as an aid in the reduction of disease due to (a specific agent)." The differences in claims and recommendations are a function of the variation in protection elicited by various vaccines. For designing effective eradication programs, vaccine efficacy characteristics such as for reducing susceptibility to infections and spread of infections must be well defined; similarly, diagnostic test performance characteristics (efficacy) must be determined. In addition to data to support efficacy claims, it is imperative that safety of production and use of vaccines be evaluated. During the design of marker vaccines and diagnostic tests, it is important to consider the application of appropriate technologies to improve the safety of these products. Use of recombinant technologies for production of vaccines and/or diagnostic test antigens can reduce the biosafety concerns during production and during use, including human exposure to zoonotic pathogens during production and use, and potential spread of foreign animal disease agents due to loss of biocontainment. In addition, vaccines may induce adverse reactions. It is important to determine the frequency of adverse events and to reduce the likelihood of induction of adverse reactions through proper design. PMID- 16257546 TI - Evidence for developmental changes in the visual word processing network beyond adolescence. AB - Late development of specialization in the visual word processing system was examined using event-related potentials (ERP) and functional magnetic resonance imaging (fMRI) of word and symbol string processing in groups of adolescents (15.2-17.3 years) and adults (19.8-30.8 years). We focused our ERP analyses on fast visual activity: the occipital P1 (82-131 ms) modulated by physical stimulus characteristics and the occipito-temporal N1 (132-256 ms) reflecting visual tuning for print. Our fMRI analyses concentrated on basal occipito-temporal activations in the visual word form area VWFA. For words, the correlation of fMRI activation in the VWFA and N1 amplitude confirmed the close relationship of the electrophysiological N1 with metabolic activity in the VWFA. Further support for this relationship came from low resolution electromagnetic tomography localizing the word-specific N1 near the VWFA. Both imaging techniques revealed age independent differences between words and symbol strings. Late development, however, was preferentially detected with ERPs. Decreases of P1 and N1 amplitudes with age were not limited to words and suggested further maturation of the underlying brain microstructure and function. Following adolescence, decreasing N1 latencies specific to words point to continued specialization of the visual word processing system. Both N1 and fMRI measures correlated with reading performance. In summary, the similarity of global fMRI activation patterns between groups suggests a fully established distribution of the reading network in adolescence, while the decreasing N1 latencies for words indicate protracted fine tuning after adolescence. PMID- 16257548 TI - Beneficial effects on sleep of vagus nerve stimulation in children with therapy resistant epilepsy. AB - The study purpose was to evaluate sleep structure following Vagus Nerve Stimulation (VNS) in 15 children with therapy resistant epilepsy and to correlate possible alterations with changes in epileptiform activity and clinical effects. Fifteen children were examined with ambulatory polysomnographic recordings initially, and after 3 and 9 months of VNS-treatment. Sleep parameters, all-night delta power activity and movement times (MTs), used to account for arousals were estimated. Epileptiform activity was evaluated by spike detection. Seizure frequency was recorded in a diary. The severity of the seizures was scored with the National Hospital Seizure Severity Scale (NHS3). Quality of life (QOL) was assessed by a visual analogue scale. Behaviour problems were quantified by using the total score of the Child Behaviour Checklist (CBCL). VNS induces a significant increase in slow wave sleep (SWS) and a decrease in sleep latency and in stage 1 sleep. The number and density of MTs during total night sleep were significantly increased. There was also a significant increase in the number of MTs immediately related to the VNS stimulation periods. Of the 14 children with increased MTs, 10 had a reduction in epileptiform activity, and in clinical seizures, all had an improvement in NHS3, and 11 in QOL. Of the 10 children with increased SWS, eight also improved in QOL and eight in behaviour. Our findings indicate that VNS counteracts known adverse effects of epilepsy on sleep and increases slow wave sleep. This possibly contributes to the reported improvement in well-being. We also see an increase in MTs. This arousal effect seems to be of minor importance for QOL and could possibly be related to the antiepileptic mechanisms in VNS. PMID- 16257547 TI - Ecphory of autobiographical memories: an fMRI study of recent and remote memory retrieval. AB - Ecphory occurs when one recollects a past event cued by a trigger, such as a picture, odor, or name. It is a central component of autobiographical memory, which allows us to "travel mentally back in time" and re-experience specific events from our personal past. Using fMRI and focusing on the role of medial temporal lobe (MTL) structures, we investigated the brain bases of autobiographical memory and whether they change with the age of memories. Importantly, we used an ecphory task in which the remote character of the memories was ensured. The results showed that a large bilateral network supports autobiographical memory: temporal lobe, temporo-parieto-occipital junction, dorsal prefrontal cortex, medial frontal cortex, retrosplenial cortex and surrounding areas, and MTL structures. This network, including MTL structures, changed little with the age of the memories. PMID- 16257549 TI - Homology modelling and protein structure based functional analysis of five cucumovirus coat proteins. AB - Coat proteins (CP) of five cucumovirus isolates, Cucumber mosaic virus (CMV) strains R, M and Trk7, Tomato aspermy virus (TAV) strain P and Peanut stunt virus (PSV) strain Er, were constructed by homology modelling. The X-ray structure of the Fny-CMV CP subunit B was used as a template. Models of cucumovirus CPs were built by the MODELLER program. Model refinements were carried out using the Kollman molecular mechanical force field. Models were analyzed by the PROCHECK programs. Electrostatic potential calculations were applied to all models and functional site search was performed with the PROSITE software, a web based tool for searching biologically significant sites. Symptom determinants published up to the present were compared with the PROSITE hits in the light of 3D models and electrostatic information. In all cases, we analyzed the effect of mutations on the structure, electrostatic potential patterns and function of CPs, respectively. We found that high flexibility of the betaE-alphaEF loop starting with the residue 129 is required, but it is not sufficient for the symptom appearance. Furthermore, phosphorylation of the CP is prospective to be important in the host response mechanism. All analyzed mutations were related to the modifications of the predicted phosphorylation sites. Based on our conclusions we predicted the infectivity of the examined viruses. PMID- 16257550 TI - Inhibition of airway hyperresponsiveness and pulmonary inflammation by roflumilast and other PDE4 inhibitors. AB - Roflumilast is an oral, once-daily phosphodiesterase 4 (PDE4) inhibitor with anti inflammatory activity. We compared the anti-inflammatory effects of roflumilast with those of PDE4 inhibitors rolipram, piclamilast, and cilomilast in ovalbumin (OVA)-sensitized and challenged Brown-Norway rats. Animals were treated orally 1h before OVA challenge with roflumilast (0.3, 1.0, and 3.0mg/kg), rolipram (0.8, 2.8, and 8.3mg/kg), piclamilast (10.0, 20.0, and 30.0mg/kg), or cilomilast (10.3, 34.3, and 103.0mg/kg). Airway hyperresponsiveness (AHR) against adenosine was investigated by measuring airway resistance 200min after OVA challenge. Subsequently, neutrophil influx and tumor necrosis factor-alpha (TNF-alpha) release in the lungs were determined by bronchoalveolar lavage. Direct bronchodilation at the time point of AHR assessment by PDE4 inhibitors was examined in serotonin-challenged animals. Evaluation of neutropenic animals or treatment with anti-TNF-alpha antibody revealed that AHR was independent of neutrophil accumulation or TNF-alpha release. Roflumilast (50% inhibitory dose [ID(50)]=1.5mg/kg) inhibited AHR 3-, 16-, and 27-fold more potently than rolipram, piclamilast, and cilomilast, respectively. Likewise, roflumilast was a more potent inhibitor of neutrophil influx (ID(50)=0.9mg/kg) than rolipram (ID(50)=6.9mg/kg), piclamilast (ID(50)=28.1mg/kg), or cilomilast (ID(50)=37.7mg/kg). Roflumilast, rolipram, and piclamilast-but not cilomilast suppressed OVA-induced TNF-alpha release in a dose-dependent manner. Roflumilast (ID(50)=0.9mg/kg) exhibited 9- and 23-fold more potent inhibition of TNF-alpha release than rolipram and piclamilast, respectively. Roflumilast did not inhibit serotonin-induced bronchoconstriction 4.5h after administration, suggesting that inhibition of AHR by roflumilast results from anti-inflammatory, not bronchodilatory, effects. This study suggests that roflumilast has anti inflammatory action and provides rationale for the investigation of roflumilast in asthmatic patients. PMID- 16257551 TI - An assessment of dead space in pulmonary ventilation of the toad Bufo schneideri. AB - The respiratory cycles of Rana and Bufo has been disputed in relation to flow patterns and to the respiratory dead-space of the buccal volume. A small tidal volume combined with a much larger buccal space motivated the "jet steam" model that predicts a coherent expired flow within the dorsal part of the buccal space. Some other studies indicate an extensive mixing of lung gas within the buccal volume. In Bufo schneideri, we measured arterial, end-tidal and intrapulmonary PCO(2) to evaluate dead-space by the Bohr equation. Dead-space was also estimated as: V(D)=(total ventilation-effective ventilation)/f(R), where total ventilation and f(R) were measured by pneumotachography, while effective ventilation was derived from the alveolar ventilation equation. These approaches were consistent with a dead space of 30-40% of tidal volume, which indicates a specific pathway for the expired lung gas. PMID- 16257552 TI - Physiological responses to hyper-saline waters in sailfin mollies (Poecilia latipinna). AB - We examined the ionoregulatory physiology and biochemistry of the teleost sailfin molly (Poecilia latipinna), an inhabitant of salt marshes along the gulf coast, during exposure to hyper-saline waters (salinity range 35-95 ppt). Mollies were able to tightly control plasma Na(+) and Cl(-) concentrations and tissue water levels up to 65 ppt, but at higher salinities plasma ion levels began to rise and muscle water content dropped. Still, even at the highest salinity (90 ppt) plasma Na(+) and Cl(-) levels were only 32% and 39%, respectively, above levels at 35 ppt. Drinking rates at 60 ppt climbed 35%, while gut Na(+)/K(+)-ATPase (NAK) activity rose 70% and branchial NAK activity jumped 200%. The relatively small rise in drinking rate, in the face of a more than doubling of the osmotic gradient, suggests that a reduction in branchial water permeability significantly limited water loss and associated salt load. At 80 ppt, a salinity where plasma ion levels just begin to rise, drinking rate rose more rapidly, but gut and gill NAK activity did not, suggesting that mollies employed other pathways (perhaps renal) of salt excretion. At higher salinities, plasma ion levels continued to rise and muscle water content fell slightly indicating the beginnings of internal osmotic disturbances. To evaluate the energetic costs of hyper-salinity on mollies we measured the rate of O(2) consumption and found it rose with salinity, in sharp contrast to virtually all species previously examined. Interestingly, despite higher metabolism, growth was unaffected by hyper-salinity. PMID- 16257553 TI - Food-deprivation induces HSP70 and HSP90 protein expression in larval gilthead sea bream and rainbow trout. AB - Heat shock proteins (HSPs) expression is commonly used as indicators of cellular stress in animals. However, very little is known about either the expression patterns of HSPs or their role in the stress-tolerance phenomenon in early life stages of fish. To this end, we examined the impact of food-deprivation (12 h), reduced oxygen levels (3.5 mg/L for 1 h) and heat shock (HS: +5 degrees C for 1 h) on HSP70 and HSP90 protein expression in early life stages of the gilthead sea bream (Sparus aurata), a warm-water aquaculture species. Also, we investigated HSP70 and HSP90 response to food-deprivation (7 days) in early life stages of rainbow trout (Oncorhynchus mykiss), a cool-water aquaculture species, and the tolerance of this larvae to heat shock (either +5 or +10 degrees C for 1 h). Our results clearly demonstrate that food-deprivation enhances HSP70 and HSP90 protein expression in larvae of both species. In gilthead sea bream larvae, the stressors-induced HSP70 and HSP90 (only in the reduced oxygen group) protein expression returned to unstressed levels after 24 h recovery. In fed trout larvae, a +5 degrees C heat shock did not elevate HSP70 and HSP90 expression, whereas 100% mortality was evident with a +10 degrees C HS. However, food deprived trout larvae, which had higher HSP70 and HSP90 protein content, survived HS and showed HS-dependent increases in HSP70, but not HSP90 expression. Overall, HSP70 and HSP90 protein expression in early life stages of fish have the potential to be used as markers of nutritional stress, while elevation of the tissue HSPs content may be used as a means to increase stress tolerance during larval rearing. PMID- 16257554 TI - Effect of dietary lipids on plasma fatty acid profiles and prostaglandin and leptin production in gilthead seabream (Sparus aurata). AB - The aim of this study was to investigate the effects of different levels of substitution of fish oil by vegetable oils rich in oleic, linoleic and linolenic acids on gilthead seabream plasma and leukocyte fatty acid compositions and prostaglandin (PG) and leptin production. Juvenile seabream of 24 g initial body mass were fed four iso-energetic and iso-proteic experimental diets for 281 days. Fatty acid composition of plasma lipids was markedly affected by the inclusion of vegetable oils (VO). ARA (arachidonate), EPA (eicosapentaenoate) and DHA (docosahexaenoate) were preferentially incorporated into polar lipids of plasma, and DHGLA (di-homogammalinoleate) accumulated with increased vegetable oil inclusion. Dietary treatments resulted in alterations of DHGLA/ARA ratios, but not ARA/EPA. ARA-derived PGE(2) production in plasma was not affected by vegetable oils, in agreement with similar eicosanoid precursor ratio (ARA/EPA) in leukocytes total lipids and plasma phospholipids among fish fed with the different dietary treatments. Feeding vegetable oils leads to a decrease in plasma EPA which in turn reduced plasma PGE(3) concentration. Moreover, PGE(3) was the major prostaglandin produced in plasma of fish fed fish oil based diet. Such findings point out the importance of EPA as a precursor of prostaglandins in marine fish, at least for the correct function of the blood cells, and correlates well with the predominant role of this fatty acid in immune regulation in this species. A negative correlation was found between plasma PGE(2) and leptin plasma concentration, suggesting that circulating levels of leptin may act as a metabolic signal modulating PGE(2) release. The present study has shown that increased inclusion of vegetable oils in diet for gilthead seabream may profoundly affect the fatty acid composition of plasma and leukocytes, specially HUFA (highly unsaturated fatty acids), and consequently the production of PGE(3), which can be a major PG in plasma. Alteration in the amount and type of PG produced can be at least partially responsible for the changes in the immune system and health parameters of fish fed diets with high inclusion of VO. PMID- 16257556 TI - Chromosomal promoter replacement of the isoprenoid pathway for enhancing carotenoid production in E. coli. AB - For metabolic engineering it is advantageous in terms of stability, genetic regulation, and metabolic burden to modulate expression of relevant genes on the chromosome rather than relying on over-expression of the genes on multi-copy vectors. Here we have increased the production of beta-carotene in Escherichia coli by replacing the native promoter of the chromosomal isoprenoid genes with the strong bacteriophage T5 promoter (P(T5)). We recombined PCR fragments with the lambda-Red recombinase to effect chromosomal promoter replacement, which allows direct integration of a promoter along with a selectable marker that can subsequently be excised by the Flp/FRT site-specific recombination system. The resulting promoter-engineered isoprenoid genes were combined by serial P1 transductions into a host strain harboring a reporter plasmid containing beta carotene biosynthesis genes allowing a visual screen for yellow color indicative of beta-carotene accumulation. Construction of an E. coli P(T5)-dxs P(T5) ispDispF P(T5)-idi P(T5)-ispB strain resulted in producing high titers (6mg/g dry cell weight) of beta-carotene. Surprisingly, over-expression of the ispB gene, which was expected to divert carbon flow from the isoprenoid pathway to quinone biosynthesis, resulted in increased beta-carotene production. We thus demonstrated that chromosomal promoter engineering of the endogenous isoprenoid pathway yielded high levels of beta-carotene in a non-carotenogenic E. coli. The high isoprenoid flux E. coli can be used as a starting strain to produce various carotenoids by introducing heterologous carotenoid genes. PMID- 16257555 TI - Isolation, characterization and kinetics of goat cystatins. AB - Two cysteine proteinase inhibitors I and II were purified from goat kidney using alkaline denaturation, ammonium sulphate fractionation, gel filtration on Sephadex G-75 and ion exchange chromatography on DEAE cellulose. The purified inhibitors were homogenous and showed a single band on SDS PAGE under reducing and non-reducing conditions with an apparent molecular mass of 67 kDa. The cystatin forms were stable in the range of pH 3-10 and up to 95 degrees C. Immunological identity with the sheep LMW kininogen was obtained suggesting that the inhibitor is closely related to kininogens. Spectral studies confirm that the inhibitors have predominantly an alpha-helical structure and undergo major conformational changes during complex formation with papain. The inhibitors had similar inhibitory activities on cysteine proteinases. Both inhibitors inhibited papain, ficin and bromelain competitively, with maximum affinity for papain. The overall lower affinity of these inhibitors to cysteine proteinases compared to other known cystatins can be attributed to the unusual N-terminal sequence where Leu is substituted by Ile. Furthermore, N-terminal sequence analysis revealed maximum homology to mammalian LMW kininogen. PMID- 16257557 TI - [Cost of quality assurance in radiotherapy: human and material requirements]. AB - In 2004, three new important laws were passed concerning radiotherapy services. The first two concerns the internal and external quality control of linear accelerators and the last concerns the role of the medical physicist, whose presence was made mandatory during the whole length of the treatments. These laws, which aim to improve the quality and the security of treatments, represent an increase in price that we have calculated, and which prevents them being implemented, as a joint study realised by the SFRO and the SFPM has shown. The cost of quality in radiotherapy requires investment in material and manpower and improvement in availability of the accelerators which entails a complete reorganisation of the services. Cost analysis is included. The difficulties in implementing these laws have also been evaluated and this evaluation already enables us to propose certain elements enabling us to go forward to globally improve the quality and security in radiotherapy. PMID- 16257559 TI - The importance of standardisation of laboratory evaluations in HIV vaccine trials. AB - It is important to evaluate HIV-1-specific immunological responses elicited by therapeutic or prophylactic vaccines using precise, standardised assays, so that the immunogenicity and putative efficacy of candidate vaccines may be compared. Different well-validated assays must be used to quantitate specific responses, to determine which particular strategies may be efficacious. PMID- 16257558 TI - The rationale behind a vaccine based on multiple HIV antigens. AB - The viral diversity of HIV-1 is likely to require a vaccine strategy that induces broad cellular and humoral anti-HIV-1 immunity. Our strategy is based on multiple HIV-1 DNA immunogens together with adjuvant recombinant granulocyte-macrophage stimulating factor. This article describes pre-clinical and clinical work preceding the initiation of clinical HIV-1 phase I/II trials. PMID- 16257560 TI - A DNA HIV-1 vaccine based on a fusion gene expressing non-structural and structural genes of consensus sequence of the A-C subtypes and the ancestor sequence of the F-H subtypes. Preclinical and clinical studies. AB - A potent DNA vaccine against HIV, combining a vector that takes advantage of the segregation and compartmentalization effect of bovine papilloma virus E2 protein with MultiHIV insert, expressing a fusion gene coding for the non-structural and structural proteins was developed and tested for immunogenicity in mice and humans. PMID- 16257561 TI - Preparing for phase II/III HIV vaccine trials in Africa. AB - Before performing phase II/III clinical trials in Africa, preliminary studies, including assessment and building up of clinical and laboratory infrastructures, estimates of human immunodeficiency virus incidence, investigation of the background immune response, and evaluation of the cross-clade immune response, need to be done. Plans and ongoing work in the context of the AIDS Vaccine Integrated Project and some preliminary data are presented. PMID- 16257562 TI - The constitutive properties of the brain parenchyma Part 1. Strain energy approach. AB - In this paper we study several constitutive equations for the brain based on the strain energy density function. We use the polynomial function and hyper-elastic Ogden model for the strain energy and include the energy dissipation by a Prony series expansion. The models are compared with known unconfined compression experimental results of the human brain tissue to obtain the best fitted model and brain mechanical parameters. Finite element simulations are also performed using the given constitutive equations, and numerical solutions match the analytical results very closely. The results are compared with other analytical and numerical calculations. PMID- 16257563 TI - Treatment delay period: the case of arsenicosis in rural Bangladesh. AB - Arsenic concentrations of tubewell water that exceed acceptable limits poses a serious health problem in Bangladesh. Many Bangladeshis are now suffering from arsenic-related diseases. The objectives of this paper are to examine the extent of delay in seeking medical treatment by victims of arsenic poisoning and to identify factors contributing to this delay. Questionnaire survey successfully administered to 663 victims living in two rural areas of Bangladesh provided the major data source for this study. Analysis of survey data reveal that median delay period was 12 months, but the delay period ranged from 1 month to 18 years. Because of this extremely large range, the mean delay period was about 22 months. The study identified time of identification of symptoms of arsenicosis as the most significant determinant of treatment delay followed by treatment sought from members of mobile medical teams, perceived threat, and level of education. Based on the study findings, it is recommended that the Bangladesh government and NGOs involved in arsenic mitigation and prevention efforts should educate individuals at risk for arsenic poisoning about the benefits of seeking early treatment. This study also recommends to continue to dispatch mobile medical teams to the arsenic impacted areas. PMID- 16257564 TI - Hepatic fibrosis and cirrhosis: the (myo)fibroblastic cell subpopulations involved. AB - Fibrosis, defined as the excessive deposition of extracellular matrix in an organ, is the main complication of chronic liver damage. Its endpoint is cirrhosis, which is responsible for significant morbidity and mortality. The accumulation of extracellular matrix observed in fibrosis and cirrhosis is due to the activation of fibroblasts, which acquire a myofibroblastic phenotype. Myofibroblasts are absent from normal liver. They are produced by the activation of precursor cells, such as hepatic stellate cells and portal fibroblasts. These fibrogenic cells are distributed differently in the hepatic lobule: the hepatic stellate cells resemble pericytes and are located along the sinusoids, in the Disse space between the endothelium and the hepatocytes, whereas the portal fibroblasts are embedded in the portal tract connective tissue around portal structures (vessels and biliary structures). Differences have been reported between these two fibrogenic cell populations, in the mechanisms leading to myofibroblastic differentiation, activation and "deactivation", but confirmation is required. Second-layer cells surrounding centrolobular veins, fibroblasts present in the Glisson capsule surrounding the liver, and vascular smooth muscle cells may also express a myofibroblastic phenotype and may be involved in fibrogenesis. It is now widely accepted that the various types of lesion (e.g., lesions caused by alcohol abuse and viral hepatitis) leading to liver fibrosis involve specific fibrogenic cell subpopulations. The biological and biochemical characterisation of these cells is thus essential if we are to understand the mechanisms underlying the progressive development of excessive scarring in the liver. These cells also differ in proliferative and apoptotic capacity, at least in vitro. All this information is required for the development of treatments specifically and efficiently targeting the cells responsible for the development of fibrosis/cirrhosis. PMID- 16257565 TI - Assembly of tight junction is regulated by the antagonism of conventional and novel protein kinase C isoforms. AB - Apparently conflicting observations indicated that protein kinase C both may block and support the assembly of tight junctions. We therefore tested the hypothesis that different isoenzymes antagonistically affect tight junction proteins and function. Thus, by using specific inhibitors we investigated the involvement of conventional and novel protein kinase C of kidney tubule cells in tight junction assembly. In low Ca2+ medium, the application of pan-protein kinase C inhibitor GF-109203X blocked the formation of tight junctions induced by protein kinase C agonist diacyglycerol. Go6976, inhibitor of conventional protein kinase C, promoted the formation of tight junctions and occludin phosphorylation in cells cultivated in low Ca2+ medium and attenuated the disruption of tight junction complex induced by the switch to low Ca2+ medium. In addition, Go6976 accelerated the occludin phosphorylation and the formation of tight junction barrier during assembly of tight junctions induced by Ca2+ re-addition. This phosphorylation was accompanied by accelerated occludin incorporation into newly forming tight junctions and by reducing the paracellular permeability. In contrast, inhibitor of novel protein kinase C rottlerin blocked the occludin phosphorylation and the formation of tight junction barrier, both caused by re addition of normal Ca2+ medium. It is concluded that the conventional protein kinase C alpha participates in tight junction disassembly while the novel protein kinase C epsilon plays a role in tight junction formation of kidney epithelial cells. The discovered antagonism contributes to a better understanding of the regulation of the structure and function of tight junctions and hence to that of the epithelial barrier. PMID- 16257567 TI - Electrostatics in computational protein design. AB - Catalytic activity and protein-protein recognition have proven to be significant challenges for computational protein design. Electrostatic interactions are crucial for these and other protein functions, and therefore accurate modeling of electrostatics is necessary for successfully advancing protein design into the realm of protein function. This review focuses on recent progress in modeling electrostatic interactions in computational protein design, with particular emphasis on continuum models. PMID- 16257566 TI - Role of anti-TGF-beta antibodies in the treatment of renal injury. AB - Chronic kidney diseases are emerging world wide as a public health problem. Finding more effective renoprotective therapies is a major challenge in nephrology. Soon after the discovery of transforming growth factor-beta (TGF beta), it became clear that this cytokine is widely expressed in almost all kidney cells. Overexpression of TGF-beta isoforms and their receptors in the kidney of experimental animals is closely linked to renal fibrosis, a characteristic feature of progressive proteinuric nephropathies. Antibodies to TGF-beta halt the development of diabetic and non-diabetic nephropathies. The effect is maximized by the addition of TGF-beta antibodies on a background of angiotensin II blockade. Several studies have pointed to a role for TGF-beta in instigating the fibrotic process characteristic of progressive human glomerulopathies, although the possible benefit of selective inhibition of TGF beta has not been described so far. This chapter discusses the role of TGF-beta in renal fibrosis, and describes the renoprotective potential of strategies that interfere with TGF-beta production in the kidney in experimental animals, as a new anti-fibrotic therapeutic approach. PMID- 16257570 TI - Recent advances in lipid molecular design. AB - The area of lipid molecular design is attracting widespread interest among numerous research groups worldwide. Diverse lipid assemblies in aqueous media, such as vesicles, bilayers and nanorods, offer new applications in chemical biology. Lipids with specifically tailored molecular architecture have been successfully employed as gene delivery vehicles, for controlled drug release and the preparation of supramolecular gels. Such molecular design of lipids, as well as their characterization upon membrane formation, offers an insight into the possible molecular basis of their properties. This in turn helps in the design of further generations of lipid systems with more predictable characteristics. Here, we present an overview of the current trends in lipid design and their utilization in various biochemical, physical and chemical applications. PMID- 16257571 TI - Molecular recognition with designed peptides and proteins. AB - The design of proteins and peptides as molecular receptors is a rapidly growing area of research. Two primary approaches have been utilized, involving the minimization of known protein binding motifs or the de novo design of binding pockets within well-folded protein structures. These approaches are complementary and help define the minimum requirements necessary for biomolecular recognition. Recent advances in this area include the design of cavities within helix bundles for the binding of anesthetics, the design of beta-hairpins for the recognition of nucleotides and oligonucleotides, the redesign of protein binding sites for unique ligands, and the design of mini-proteins via protein grafting for the recognition of proteins and DNA. These advances provide exciting new opportunities to develop novel biosensors, de novo designed catalysts, exogenously triggered synthetic signal transduction cascades, and novel approaches to therapeutic treatments. PMID- 16257572 TI - Protein components for nanodevices. AB - A long-term goal of nanobiotechnology is to build tiny devices that respond to the environment, perform computations and carry out tasks. Considerable progress has been made in building protein components for such devices, and here we describe examples, including self-assembling protein arrays, pores with triggers and switches, and motor proteins harnessed for specific tasks. A major issue that has been successfully addressed in this recent work is the interface between the proteins and other components of the system, such as a metal surface. While further progress is expected in the coming years, the assembly of devices from the components has seen more limited accomplishments. For example, although a wide variety of sensors based on nanobiotechnology has been developed, unresolved problems still confront the construction of complex nanobioelectronic circuits, and the development of nanorobotics with biological components remains a distant dream. PMID- 16257573 TI - Genetic technologies for Archaea. AB - Members of the third domain of life, the Archaea, possess structural, physiological, biochemical and genetic features distinct from Bacteria and Eukarya and, therefore, have drawn considerable scientific interest. Physiological, biochemical and molecular analyses have revealed many novel biological processes in these important prokaryotes. However, assessment of the function of genes in vivo through genetic analysis has lagged behind because suitable systems for the creation of mutants in most Archaea were established only in the past decade. Among the Archaea, sufficiently sophisticated genetic systems now exist for some thermophilic sulfur-metabolizing Archaea, halophilic Archaea and methanogenic Archaea. Recently, there have been developments in genetic analysis of thermophilic and methanogenic Archaea and in the use of genetics to study the physiology, metabolism and regulatory mechanisms that direct gene expression in response to changes of environmental conditions in these important microorganisms. PMID- 16257574 TI - Cytogenetic analysis of nasal mucosa cells and lymphocytes from high-level long term formaldehyde exposed workers and low-level short-term exposed waiters. AB - The evidence for genotoxic potential of formaldehyde (FA) in humans is insufficient and conflicting. We previously reported a higher frequency of micronuclei in nasal and oral exfoliative cells from students exposed to formaldehyde vapor for short-term. To further evaluate the genetic effects of long-term occupational exposure to FA and short-term exposure to FA of indoor sources, the frequencies of micronuclei (MN) in nasal mucosa cells, sister chromatid exchanges (SCEs) of peripheral lymphocytes, and the lymphocyte subsets were evaluated in 18 non-smoking workers (mean exposure duration was 8.6 years) in an FA factory and 16 non-smoking waiters exposed to FA for 12 weeks in a ballroom. A non-smoking student group without occupational exposure (n=23) to FA was used as control. The 8h time-weighted average (TWA) concentrations of formaldehyde was 0.985+/-0.286 mg/m3 with the ceiling exposure concentration of 1.694 mg/m3 in the workshop, and 0.107+/-0.067 mg/m3 in the ballroom (5 h TWA). Higher frequencies of micronuclei per thousand cells in nasal mucosa cells of workers versus control (2.70+/-1.50 versus 1.25+/-0.65, p<0.05) and higher frequency of SCEs in peripheral lymphocytes of workers group (8.24+/-0.89 versus 6.38+/-0.41, p<0.05) were observed. Increased frequency of micronuclei in nasal mucosa cells or SCE in peripheral lymphocytes was not found among waiters group. The results suggest that the genotoxic potential of high level FA exposure may have occupational risks in long-term exposure groups. PMID- 16257575 TI - In silico assessment of chemical mutagenesis in comparison with results of Salmonella microsome assay on 909 chemicals. AB - Genotoxicity is one of the important endpoints for risk assessment of environmental chemicals. Many short-term assays to evaluate genotoxicity have been developed and some of them are being used routinely. Although these assays can generally be completed within a short period, their throughput is not sufficient to assess the huge number of chemicals, which exist in our living environment without information on their safety. We have evaluated three commercially available in silico systems, i.e., DEREK, MultiCASE, and ADMEWorks, to assess chemical genotoxicity. We applied these systems to the 703 chemicals that had been evaluated by the Salmonella/microsome assay from CGX database published by Kirkland et al. We also applied these systems to the 206 existing chemicals in Japan that were recently evaluated using the Salmonella/microsome assay under GLP compliance (ECJ database). Sensitivity (the proportion of the positive in Salmonella/microsome assay correctly identified by the in silico system), specificity (the proportion of the negative in Salmonella/microsome assay correctly identified) and concordance (the proportion of correct identifications of the positive and the negative in Salmonella/microsome assay) were increased when we combined the three in silico systems to make a final decision in mutagenicity, and accordingly we concluded that in silico evaluation could be optimized by combining the evaluations from different systems. We also investigated whether there was any correlation between the Salmonella/microsome assay result and the molecular weight of the chemicals: high molecular weight (>3000) chemicals tended to give negative results. We propose a decision tree to assess chemical genotoxicity using a combination of the three in silico systems after pre-selection according to their molecular weight. PMID- 16257576 TI - Two subtypes (subgenotypes) of hepatitis B virus genotype C: A novel subtyping assay based on restriction fragment length polymorphism. AB - Recently hepatitis B virus genotype C (HBV/C) has been classified into geographically typical two subtypes (subgenotypes); HBV/C1 in Southeast Asia (Cs) and HBV/C2 in East Asia (Ce). Our aim is to develop a rapid subtyping assay and to examine the virological features of these two subtypes. Based on 171 HBV/C strains retrieved from the database, 17 single nucleotides polymorphisms (SNPs) were found between two subtypes. Taking advantage of five SNPs in non-overlapping polymerase region, a restriction fragment length polymporphism method with three endonucleases was newly developed for distinguishing between HBV/Cs and HBV/Ce. The method was applied to 49 HBV/C carriers from Japan and Hong Kong. The 24 in Hong Kong were classified into HBV/Cs, and the 25 in Japan were HBV/Ce, confirmed by sequencing. Some specific mutations were detected in the encapsidation signal; precore stop mutation (A1896), accompanied by a C-to-T substitution at nt 1858, was found in HBV/Ce strains, and another precore mutation (A1898), accompanied by a C-to-T mutation at nt 1856, was found in HBV/Cs. Especially, two closely linked mutations (A1896 and A1899) in HBV/Ce could stabilize the epsilon loop structure more efficiently and influece viral replication. Hence, these virological differences between the two subtypes might influence clinical features. PMID- 16257577 TI - Reference-free quantification of EEG spectra: combining current source density (CSD) and frequency principal components analysis (fPCA). AB - OBJECTIVE: Definition of appropriate frequency bands and choice of recording reference limit the interpretability of quantitative EEG, which may be further compromised by distorted topographies or inverted hemispheric asymmetries when employing conventional (non-linear) power spectra. In contrast, fPCA factors conform to the spectral structure of empirical data, and a surface Laplacian (2 dimensional CSD) simplifies topographies by minimizing volume-conducted activity. Conciseness and interpretability of EEG and CSD fPCA solutions were compared for three common scaling methods. METHODS: Resting EEG and CSD (30 channels, nose reference, eyes open/closed) from 51 healthy and 93 clinically-depressed adults were simplified as power, log power, and amplitude spectra, and summarized using unrestricted, Varimax-rotated, covariance-based fPCA. RESULTS: Multiple alpha factors were separable from artifact and reproducible across subgroups. Power spectra produced numerous, sharply-defined factors emphasizing low frequencies. Log power spectra produced fewer, broader factors emphasizing high frequencies. Solutions for amplitude spectra showed optimal intermediate tuning, particularly when derived from CSD rather than EEG spectra. These solutions were topographically distinct, detecting multiple posterior alpha generators but excluding the dorsal surface of the frontal lobes. Instead a low alpha/theta factor showed a secondary topography along the frontal midline. CONCLUSIONS: CSD amplitude spectrum fPCA solutions provide simpler, reference-independent measures that more directly reflect neuronal activity. SIGNIFICANCE: A new quantitative EEG approach affording spectral components is developed that closely parallels the concept of an ERP component in the temporal domain. PMID- 16257578 TI - Cost-effective production of 13C, 15N stable isotope-labelled biomass from phototrophic microalgae for various biotechnological applications. AB - The present study outlines a process for the cost-effective production of 13C/15N labelled biomass of microalgae on a commercial scale. The core of the process is a bubble column photobioreactor with exhaust gas recirculation by means of a low pressure compressor. To avoid accumulation of dissolved oxygen in the culture, the exhaust gas is bubbled through a sodium sulphite solution prior to its return to the reactor. The engineered system can be used for the production of 13C, 15N, and 13C-15N stable isotope-labelled biomass as required. To produce 13C-labelled biomass, 13CO2 is injected on demand for pH control and carbon supply, whereas for 15N-labelled biomass Na15NO3 is supplied as nitrogen source at the stochiometric concentration. The reactor is operated in semicontinuous mode at different biomass concentrations, yielding a maximum mean biomass productivity of 0.3 gL(-1) day(-1). In order to maximize the uptake efficiency of the labelled substrates, the inorganic carbon is recovered from the supernatant by acidification/desorption processes, while the nitrate is delivered at stochiometric concentration and the harvesting of biomass is performed when the 15NO3- is depleted. In these conditions, elemental analysis of both biomass and supernatant shows that 89.2% of the injected carbon is assimilated into the biomass and 6.9% remains in the supernatant. Based on elemental analysis, 97.8% of the supplied nitrogen is assimilated into the biomass and 1.3% remains in the supernatant. Stable isotope-labelling enrichment has been analysed by GC-MS results showing that the biomass is highly labelled. All the fatty acids are labelled; more than 96% of the carbon present in these fatty acids is 13C. The engineered system was stably operated for 3 months, producing over 160 g of 13C and/or 15N-labelled biomass. The engineered bioreactor can be applied for the labelling of various microalgae. PMID- 16257579 TI - Human Mycobacterium bovis infection in the United Kingdom: Incidence, risks, control measures and review of the zoonotic aspects of bovine tuberculosis. AB - Amongst the members of the Mycobacterium tuberculosis complex (MTBC), M. tuberculosis is mainly a human pathogen, whereas M. bovis has a broad host range and is the principal agent responsible for tuberculosis (TB) in domestic and wild mammals. M. bovis also infects humans, causing zoonotic TB through ingestion, inhalation and, less frequently, by contact with mucous membranes and broken skin. Zoonotic TB is indistinguishable clinically or pathologically from TB caused by M. tuberculosis. Differentiation between the causative organisms may only be achieved by sophisticated laboratory methods involving bacteriological culture of clinical specimens, followed by typing of isolates according to growth characteristics, biochemical properties, routine resistance to pyrazinamide (PZA) and specific non-commercial nucleic acid techniques. All this makes it difficult to accurately estimate the proportion of human TB cases caused by M. bovis infection, particularly in developing countries. Distinguishing between the various members of the MTBC is essential for epidemiological investigation of human cases and, to a lesser degree, for adequate chemotherapy of the human TB patient. Zoonotic TB was formerly an endemic disease in the UK population, usually transmitted to man by consumption of raw cows' milk. Human infection with M. bovis in the UK has been largely controlled through pasteurization of cows' milk and systematic culling of cattle reacting to compulsory tuberculin tests. Nowadays the majority of the 7000 cases of human TB annually reported in the UK are due to M. tuberculosis acquired directly from an infectious person. In the period 1990-2003, between 17 and 50 new cases of human M. bovis infection were confirmed every year in the UK. This represented between 0.5% and 1.5% of all the culture-confirmed TB cases, a proportion similar to that of other industrialized countries. Most cases of zoonotic TB diagnosed in the UK are attributed to (i) reactivation of long-standing latent infections acquired before widespread adoption of milk pasteurization, or (ii) M. bovis infections contracted abroad. Since 1990, only one case has been documented in the UK of confirmed, indigenous human M. bovis infection recently acquired from an animal source. Therefore, for the overwhelming majority of the population, the risk of contracting M. bovis infection from animals appears to be extremely low. However, bovine TB is once again a major animal health problem in the UK. Given the increasing numbers of cattle herds being affected each year, physicians and other public health professionals must remember that zoonotic TB is not just a disease of the past. A significant risk of M. bovis infection remains in certain segments of the UK population in the form of (i) continuing on-farm consumption of unpasteurized cows' milk, (ii) retail sales by approved establishments of unpasteurized milk and dairy products and (iii) occupational exposure to infectious aerosols from tuberculous animals and their carcases. PMID- 16257580 TI - Reliability of clinical muscular endurance tests in patients with chronic heart failure. AB - BACKGROUND: In clinical praxis it is important to be able to assess the effect of rehabilitation in patients with chronic heart failure (CHF). There is therefore a need for simple clinical tests that are objective, reliable and valid measures. AIM: The aim of this study was to examine the reliability of three muscle endurance tests for patients with CHF and to compare the results to a healthy control group. METHOD: The study included 20 patients (7 women) with CHF and 20 healthy persons. Dynamic endurance of the shoulder and calf muscle was measured as well as static shoulder endurance. All three tests were performed twice, within 5-10 days. RESULTS: The reliability (rs) of the tests for the patients with CHF was between 0.90-0.99 and for the healthy persons between 0.79-0.98. There was also a significant difference in dynamic endurance in shoulder flexion and in heel-rise left leg test between patients with CHF and healthy persons. CONCLUSION: Tests of dynamic and static endurance of the shoulder muscle and dynamic calf muscle endurance are reliable assessment tools for use in clinical practice. Patients with CHF have reduced isotonic muscular endurance compared with healthy persons. PMID- 16257581 TI - The role and applications of ultrasound in ambulatory gynaecology. AB - Ultrasound has changed gynaecological practice and continues to do so. One of the earliest applications of abdominal scanning in gynaecology was for monitoring follicular development during fertility treatment with clomiphene citrate or gonadotrophins in the 1960s and 1970s. Subsequently, it was natural that with the introduction of in vitro fertilization, abdominal and transvaginal ultrasound played a key role in the development of oocyte retrieval techniques. These were truly the first interventional ultrasound-guided ambulatory procedures in gynaecology. In this chapter, the reader will be introduced to the roles that the various ultrasound modalities play in our current daily practice, and how they have changed the management of numerous gynaecological conditions in both diagnostic and therapeutic contexts. We will also outline the recent developments and the 'hot' research topics in this field. PMID- 16257583 TI - Analysis of CpG methylation in the killifish CYP1A promoter. AB - Fundulus heteroclitus (Atlantic killifish or mummichog) inhabiting a creosote contaminated Superfund site on the Elizabeth River (VA, USA), exhibit a lack of induction of cytochrome P4501A (CYP1A) mRNA, immunodetectable protein, and catalytic activity after exposure to typical inducers. This "refractory CYP1A phenotype" is not explained by alterations in mRNA expression of known CYP1A transcription factors. Furthermore, the refractory phenotype is lost progressively during development in laboratory-reared F1 generation fish. Thus, while heritable, the refractory CYP1A phenotype does not appear to be genetically based. To test the hypothesis that cytosine methylation at CpG sites in the promoter region of CYP1A underlies the refractory CYP1A phenotype, we employed bisulfite sequencing to compare the methylation status of CpG sites in the CYP1A promoter region of DNA from killifish from the Elizabeth River and a reference site. We examined genomic DNA both from livers of wild-caught adult killifish and from pools of F1 generation embryos raised in the laboratory. In fish from both the contaminated and the reference site, cytosine methylation was not detectable at any of the 34 CpG sites examined, including 3 that are part of putative xenobiotic response elements. PMID- 16257582 TI - Phase I/II trial of intravenous NDV-HUJ oncolytic virus in recurrent glioblastoma multiforme. AB - We undertook a Phase I/II trial in patients with apparent recurrent glioblastoma multiforme (GBM) based on imaging studies to determine the safety and tumor response of repetitive intravenous administration of NDV-HUJ, the oncolytic HUJ strain of Newcastle disease virus. The first part of the study utilized an accelerated intrapatient dose-escalation protocol with one-cycle dosage steps of 0.1, 0.32, 0.93, 5.9, and 11 billion infectious units (BIU) of NDV-HUJ (1 BIU = 1 x 10(9) EID(50) 50% egg infectious dose) followed by three cycles of 55 BIU. Virus was administered by intravenous infusion over 15 min. In the second part, patients received three cycles of 11 BIU. All patients without progressive disease were maintained with two doses of 11 BIU iv weekly. Eleven of the 14 enrolled patients (11-58 years, Karnofsky performance scale 50-90%) received treatment. Toxicity was minimal with Grade I/II constitutional fever being seen in 5 patients. Maximum tolerated dose was not achieved. Anti-NDV hemagglutinin antibodies appeared within 5-29 days. NDV-HUJ was recovered from blood, saliva, and urine samples and one tumor biopsy. One patient achieved a complete response. Intravenous NDV-HUJ is well tolerated. The findings of good tolerability and encouraging responses warrant the continued evaluation of NDV-HUJ in GBM, as well as other cancers. PMID- 16257584 TI - Joint effects of dimethoate and heavy metals on metabolic responses in a grasshopper (Chorthippus brunneus) from a heavy metals pollution gradient. AB - We studied how an exposure to an additional stressing factor-dimethoate, might affect detoxifying ability of grasshoppers collected at 5 meadow sites located along a heavy metal pollution gradient. Activities of esterases and enzymes linked with glutathione (GSH) metabolism were assayed 24 h after topical treatment with 0.32 microg dimethoate per insect. Inhibition of acetylcholinesterase (AChE) reaches nearly 50% of the value stated in untreated insects, without significant site-dependent differences. The pesticide also caused a significant decrease in activities of glutathione peroxidase (GPx) followed by a decrease in GSH levels in grasshoppers from all assayed groups, demonstrating high sensitivity of glutathione-dependent metabolism to the additional stressing factor. In the case of glutathione reductase (GR) and carboxylesterases (CarE) the fall of activity was shown especially in insects from less polluted meadows and the reference site. Glutathione reductase (GR) activity in individuals treated with dimethoate did not decrease only in insects from the most contaminated site I. This might suggest the trade-off mechanisms adapting grasshoppers to life in seriously polluted environments. PMID- 16257585 TI - Baseline characteristics of menopausal women with coronary heart disease in cardiovascular outcomes clinical trials of menopausal hormone therapy or raloxifene. AB - Despite differences in enrollment criteria, the baseline characteristics of the menopausal women with coronary heart disease enrolled in the cardiovascular outcome trials of menopausal hormone therapy or raloxifene are remarkably similar. Globally, cardiovascular risk factors were not optimally controlled at entry into these trials. More aggressive cardiovascular risk factor interventions are requisite to achieve optimal target goals for women with documented coronary heart disease. PMID- 16257586 TI - mRNA methods used in dissecting gene expression of the brain. PMID- 16257587 TI - Dale Mosbaugh commemorative DNA repair issue. PMID- 16257588 TI - Inactivation of recG stimulates the RecF pathway during lesion-induced recombination in E. coli. AB - Lesions that transiently block DNA synthesis generate replication intermediates with recombinogenic potential. In order to investigate the mechanisms involved in lesion-induced recombination, we developed an homologous recombination assay involving the transfer of genetic information from a plasmid donor molecule to the Escherichia coli chromosome. The replication blocking lesion used in the present assay is formed by covalent binding of the carcinogen N-2 acetylaminofluorene to the C8 position of guanine residues (G-AAF adducts). The frequency of recombination events was monitored as a function of the number of lesions present on the donor plasmid. These DNA adducts are found to trigger high levels of homologous recombination events in a dose-dependent manner. Formation of recombinants is entirely RecA-dependent, the RecF and RecBCD sub-pathways accounting for about 2/3 and 1/3, respectively. Inactivation of recG stimulates recombinant formation about five-fold. In a recG background, the RecF pathway is stimulated about four-fold, while the contribution of the RecBCD pathway remains constant. In addition, in the recG strain, a recombination pathway that accounts for about 30% of the recombinants and requires genes that belong to both RecF and RecBCD pathways is revealed. PMID- 16257589 TI - Validation of HPLC analysis method of a novel antihypertensive agent MS23 in rat plasma. AB - MS23 is a vasodilator with unique dual action pharmacological profile to inhibit type 4 PDE and antagonize L-type calcium channels. We validated an analytical protocol for MS23 in rat plasma using high performance liquid chromatography (HPLC). A C18 column and a phosphate/acetonitrite buffer were used for chromatographic separation. UV detection was performed at 307 nm. The calibration curve for MS23 was linear in the range from 50 to 10,000 ng/ml. The limit of quantification (LOQ) was 50 ng/ml. The results demonstrate that the method has linearity (R = 0.9989), specificity, and acceptable precision/accuracy. This method is simple, economic, and sufficient for in vivo pharmacokinetic studies on the compound. PMID- 16257590 TI - LC/MS/MS measurement of penicillin G in bovine plasma, urine, and biopsy samples taken from kidneys of standing animals. AB - Methods for the measurement of penicillin concentration in bovine plasma, kidney and urine were developed and validated. Detection was based on liquid chromatography/tandem mass spectrometry (LC/MS/MS). Phenethecillin was used as an internal standard. Plasma was extracted with acetonitrile using a method with a calculated limit of quantitation (LOQ) of 12 ng/mL. Kidney samples were homogenized in water and acetonitrile, then cleaned up on C18-bonded silica SPE cartridges. The LOQ of this procedure was 10 ng/g. Urine samples were diluted, filtered, and analyzed directly. The LOQ of this procedure was 63 ng/mL. The overall accuracy for plasma was 103% with coefficient of variation (CV) of 3%; for kidney, 96% and 11%, respectively, and for urine, 98% and 4%, respectively. These methods were applied to the analysis of plasma, urine, and kidney biopsy samples taken from standing animals that had been dosed with penicillin. PMID- 16257591 TI - Structural characterization of novel chitin-binding lectins from the genus Artocarpus and their antifungal activity. AB - Two novel chitin-binding lectins from seeds of Artocarpus genus were described in this paper, one from A. integrifolia (jackfruit) and one from A. incisa (breadfruit). They were purified from saline crude extract of seeds using affinity chromatography on chitin column, size-exclusion chromatography and reverse-phase chromatography on the C-18 column. Both are 14 kDa proteins, made up of 3 chains linked by disulfide bonds. The partial amino acid sequences of the two lectins showed they are homologous to each other but not to other plant chitin-binding proteins. Thus, they cannot be classified in any known plant chitin-binding protein family, particularly because of their inter-chain covalent bonds. Their circular dichroism spectra and deconvolution showed a secondary structure content of beta-sheet and unordered elements. The lectins were thermally stable until 80 degrees C and structural changes were observed below pH 6. Both lectins inhibited the growth of Fusarium moniliforme and Saccharomyces cerevisiae, and presented hemagglutination activity against human and rabbit erythrocytes. These lectins were denoted jackin (from jackfruit) and frutackin (from breadfruit). PMID- 16257592 TI - PRP modulates expression of bone matrix proteins in vivo without long-term effects on bone formation. AB - This experimental study (domestic pig) examined the bone formation after filling defined defects of the frontal skull with autogenous bone or a deproteinized bovine bone matrix (DBBM) in combination with platelet-rich plasma (PRP). Six groups, both materials with and without PRP in two different concentrations (4.1x and 6.5x referring to untreated whole blood) were evaluated at 2, 4, 12, and 26 weeks by means of immunohistochemical staining for different bone matrix proteins, microradiography, light microscopy and polychromatic fluorescence labeling. The sequential expression of bone matrix proteins reflected the specific roles these proteins fulfil in the mineralization of hard tissue. Collagen I expression at 2 weeks was enhanced in all autogenous bone groups. No specific modification of the collagen I expression was found after use of DBBM with or without PRP. Osteopontin and especially osteonectin showed a remarkable enhancement at 4 weeks in nearly all autogenous bone and DBBM groups. These increased levels closely resembled the mineralization content evaluated by microradiography at that time. For the three autogenous bone groups, an expression peak for osteocalcin was demonstrated at 12 weeks, further reflecting the way of de novo bone formation. The microradiographic evaluation demonstrated a statistically significant enhancement in bone regeneration by PRP only after use of autogenous bone plus PRP at 2 weeks (P = 0.002). After 4 weeks, mineralization values after use of autogenous bone were significantly lower if PRP was added to the autogenous bone (P = 0.002). No long-term effects of the PRP administration were found in the mineralization process. In all DBBM groups, bone formation remained unchanged, confirming the lack of any osteoinductive capacity of PRP. PRP modulated the expression of bone matrix proteins in this experimental setting. However, an enhancement of bone formation was demonstrated only at 2 weeks after application of the higher PRP concentration in combination with autogenous bone. In conjunction with an anorganic bovine bone no effects of PRP on defect mineralization were discovered, demonstrating the lack of osteoinductive capacity in PRP as well as in DBBM. PMID- 16257594 TI - Perinatal asphyxia pathophysiology in pig and human: a review. AB - In utero fetuses are evidently exposed to several factors that cause an interruption of the oxygen flow through the umbilical cord causing asphyxia leading to hypoxia and metabolic acidosis. These conditions are important causes of intra-partum and neonatal mortality. The main objective of this review is to provide current information regarding the pathophysiology of asphyxia in piglets around parturition; the physiological mechanisms invoked by affected piglets to compensate perinatal hypoxemia are discussed. This review also addresses some similarities and differences of asphyxia between piglets and other mammals, including human neonates. Metabolic acidosis and hypoxia are sequela to asphyxia and can cause profound health effects in postnatal performance because of an abnormal suckling, a reduced absorption of colostrum and inadequate passive transfer of neonatal immunity. Acidosis also cause hypothermia, increased mortality and reduced survival in neonates. One of the first deleterious effects of intrauterine hypoxia is the expulsion of meconium into the amniotic sac leading to meconium staining of the skin, and in severe cases, meconium aspiration into the lungs. Even though there have been technological changes and improvements in husbandry, piglet mortality due to asphyxia remains a major problem. One potential alternative to reduce neonatal mortality in pigs is the monitoring of fetal stress during birth and the implemention of strategies such as the Apgar score, that is often used in human pediatrics. It is also important to consider the physiological, behavioral and biochemical changes that take place during parturition which subsequently impact the vitality, maturity and development of neonatal pigs. Understanding the pathophysiology of fetal hypoxia should help practitioners and farmers implement more effective delivery techniques aimed at reducing neonatal mortality and improving postnatal performance. PMID- 16257593 TI - Membrane-bound peptides mimicking transmembrane Vph1p helix 7 of yeast V-ATPase: a spectroscopic and polarity mismatch study. AB - The V-ATPases are a family of ATP-dependent proton pumps, involved in a variety of cellular processes, including bone breakdown. V-ATPase enzymes that are too active in the latter process can result in osteoporosis, and inhibitors of the enzyme could be used to treat this disease. As a first step in studying the structure and function of the membrane-embedded interface at which proton translocation takes place, and its role in V-ATPase inhibition, synthetic peptides P1 and P2 consisting of 25 amino acid residues are presented here that mimic Vph1p helix 7 of yeast V-ATPase. A single mutation R10A between peptide P1 and P2 makes it possible to focus on the role of the essential arginine residue R735 in proton translocation. In the present work, we use a novel combination of spectroscopic techniques, such as CD spectroscopy, tryptophan emission spectra, acrylamide quenching and parallax analysis, and polarity mismatch modeling to characterize the peptides P1 and P2 in lipid bilayer systems. Based on both the spectroscopic experiments and the polarity mismatch modeling, P1 and P2 adopt a similar transmembrane conformation, with a mainly alpha-helical structure in the central part, placing the tryptophan residue at position 12 at a location 4+/-2 A from the centre of the lipid bilayer. Furthermore, the arginine at position 10 in P1 does not have an effect on the bilayer topology of the peptide, showing that the long, flexible side chain of this residue is able to snorkel towards the lipid headgroup region. This large flexibility of R735 might be important for its function in proton translocation in the V-ATPase enzyme. PMID- 16257595 TI - Ultrasonographic screening of embryo development in cattle (Bos indicus) between days 20 and 40 of pregnancy. AB - Through a transrectal ultrasonography (Aloka SSD 500 microplus equipment, linear transducer of 7.5 MHz) a total of 267 observations were performed in a group of 50 females mainly Bos indicus, raised under humid tropical conditions and pregnant from day 20 every other day until day 40 post-artificial insemination (AI). The objective was to measure the size of the amniotic vesicle and the longitudinal and transversal axis of the embryo, to determine the day in which it is possible to measure this structure and detect the heart contractions in the screened embryos. The accuracy in identifying the amniotic vesicle was 60% (12/20) on day 20 of the first screening with embryos having an average length of 0.44 +/- 0.20 cm. Accuracy in detecting the embryo was 66.6 and 100% between days 20 and 24 post-breeding, respectively. The mean length of the embryos on day 20 was of 0.20 +/- 0.07 cm and the transversal shape had a mean length of 0.07 +/- 0.02 cm. The accuracy of detecting heart contractions was 50% (9/18) on day 24 post-breeding and for day 26 the accuracy was 100%. At the end of the screening (day 40), embryos had means of 1.92 +/- 0.24 (longitudinal) and 0.85 +/- 0.11 cm (transverse). The embryo can be detected from day 24 post-AI and the heart contractions from day 26 with 100% reliability. PMID- 16257596 TI - Effect of artificial insemination on submission rates of lactating dairy cows synchronised and resynchronised with intravaginal progesterone releasing devices and oestradiol benzoate. AB - This study investigated the hypothesis that a reduction in submission rates at a resynchronised oestrus is not due to the resynchrony treatment involving intravaginal progesterone releasing devices (IVDs) and oestradiol benzoate (ODB) but is associated with artificial insemination (AI) at the first synchronised oestrus. In Experiment 1, cows were synchronised for first oestrus with IVDs, with ODB administered at the time of device insertion (Day 0, 2 mg IM) and 24 h after removal (Day 9, 1 mg IM) and PGF(2alpha) injected at the time of device removal. Cows were then either inseminated (I) for 4 days or not inseminated (NI) following detection of oestrus (first round of AI). Every animal was resynchronised for a second round of AI by reinsertion of IVDs on Day 23 with administration of ODB (1 mg IM) at the time of insertion as well as 24 h after removal (Day 32). Cows detected in oestrus and inseminated for 4 days at the second round of AI were resynchronised for a third round by repeating the resynchrony treatment starting on Day 46 and inseminating cows on detection of oestrus for 4 days. In Experiment 2 the same oestrous synchronisation and resynchronisation treatments were used, but the timing of treatments differed. The cows had their cycles either presynchronised (treatment start Day -23) without AI and then resynchronised, starting on Day 0, for the first round of AI for AI at detected oestrus for 4 days, or they were synchronised (treatment start Day 0) for the first round of AI. In Experiment 1, 91.4% (64/70) and 92.6% (63/68) (P = 0.79) of cows in the I and NI treatments, respectively, were detected in oestrus after the initial synchronisation. At the second round of AI, submission rates for insemination were lower in the I group compared to the NI cows (74.5%, 35/47 versus 92.6%, 63/68, respectively; P = 0.007). Pregnancy rates (proportion treated that were classified as becoming pregnant) in I and NI cows 4 weeks (61.4%, 43/70 versus 63.2%, 43/68) and 7 weeks (77.1%, 54/70 versus 69.1%, 47/68) after the AI start date (AISD) did not differ significantly between treatments. In Experiment 2, presynchronisation and then resynchronisation of oestrous cycles before the first round of AI did not affect oestrous detection rates at the first round of AI (100%, 44/44 versus 98.0%, 50/51; P = 0.54), or pregnancy rates 1 week (63.6%, 28/44 versus 60.8%, 31/51; P = 0.70), 4 weeks (72.7%, 32/44 versus 76.5%, 39/51; P = 0.76) and 7 weeks (81.8%, 36/44 versus 88.2%, 45/51; P = 0.40) after AISD compared to cows that had their cycles synchronised for the first round of AI. These findings support our hypothesis that a reduction in submission rates at a resynchronised oestrus is associated with AI at the first synchronised oestrus and not due to a resynchrony treatment involving IVDs and ODB. This study supports the concept that early embryonic loss following AI at a synchronised oestrus could cause a reduction in submission rates following resynchronisation of oestrus, although investigation of the effect of passing an AI catheter or semen components were not studied per se. PMID- 16257597 TI - Seasonal effects of semen collection and artificial insemination on dairy cow conception. AB - The effects of four seasons of semen collection and of artificial insemination on conception in dairy cows were studied. The solstices and equinoxes (December, March, June and September) defined the beginning and/or end of each season. Semen was collected from 973 progeny-test bulls over 8 years at the two Norwegian AI stations at 60.8 degrees N and 63.4 degrees N where artificial light was used to provide a minimum photoperiod of 10 h/day. The effect of using semen of elite bulls during progeny testing and after selection as elite sires also was investigated. Norwegian Red (NRF) cows were inseminated over a 7-year period using progeny test semen and over the last 4 years of the same period using the semen of the elite sires. The probability of conception to only first inseminations for cows up to, and including, the fifth lactation was assessed by 56-day non-return rate (56d NRR) and calving rate. Two data sets were analysed which excluded cows culled within 270 days of AI or included such cows as non calving. The reasons for culling were categorised as those for fertility problems or all other reasons. Semen was used for AI irrespective of the season in which it had been collected. Season of semen collection did not affect 56d NRR but calving rate was significantly higher (by 0.5-0.8%, approximately; P < 0.01) for semen collected in the December-March period, when photoperiod was increasing, than at other times of the year. The season in which AI was performed showed a peak of 56d NRR in spring for heifers (P < 0.01) and in summer for parous animals (P < 0.01). For calving rate, however, no seasonal peak was found in heifers, whereas pluriparous cows had much higher calving rates in summer and autumn/early winter than late winter and spring (P < 0.01). Semen of elite sires resulted in higher calving rates by 0.5 (NS) to 1.9% (P < 0.01) when used after selection than when used during progeny testing. The difference between the calving rate achieved when the semen from elite sires was used during progeny testing and after selection indicates that farmers select different classes of cows for submission to AI by progeny test bulls and sires. The 56d NRR was not as good as calving rate for assessing seasonal and other effects on conception rates. PMID- 16257598 TI - Validation of a 13,14-dihydro-15-keto-PGF(2alpha) enzymeimmunoassay and its application for reproductive health monitoring in postpartum buffaloes. AB - The objective of the present study was to validate a simple, sensitive and direct enzymeimmunoassay (EIA) procedure for 13,14-dihydro-15-keto-PGF(2alpha) (PGFM) for use in buffaloes with postpartum reproductive disorders and determine the practicalities of using plasma concentrations of 13,14-dihydro-15-keto PGF(2alpha) for monitoring their reproductive health. The EIA was used for determination of the circulating levels of PGFM associated with the retention of fetal membranes, postpartum endometritis and variable postpartum intervals. The concentrations of PGFM with retention of fetal membranes in the periparturient period were lower as compared to buffaloes that had uneventful parturitions. Concentrations of PGFM associated with postpartum endometritis were elevated as compared to those in buffaloes free of reproductive tract infections. Buffaloes having higher plasma concentrations of PGFM in early postpartum period had shorter postpartum intervals, indicating the association between PGFM concentrations postpartum and uterine involution as well as the resumption of estrous cycle in this species. The study presents the possibility of using circulating PGFM concentrations for monitoring the postpartum reproductive health of buffaloes. PMID- 16257599 TI - Relationships between backfat depth and plasma leptin during lactation and sow reproductive performance after weaning. AB - The aim of this study was to determine relationships between sow backfat depth, plasma leptin concentrations, and reproductive performance after weaning. On the day of farrowing (day 0), and at weaning (day 21), single blood samples were obtained from 120 mixed-parity sows and their backfat depth (P2) measured. Based on backfat depth at day 0, sows were classified as FAT (>24 mm, n = 16), MEDIUM (16-24 mm, n = 54), or THIN (<16 mm, n = 14). Sows were further classified on the basis of P2 backfat changes during lactation of <2 mm, 2-4 mm, or >4 mm. Reproductive performance was measured as weaning-to-oestrous intervals (WOI) of <6 d, 6-9 days, or > or =10 d, and pregnancy rates. There was a positive relationship (P < 0.0001) between backfat depth at day 0 and backfat loss during lactation. The WOI was not associated with backfat depth at day 0 or 21 (P > 0.1 for both). Pregnancy rate was not associated with backfat depth at day 0 (P > 0.1) but pregnant sows had a greater backfat depth at weaning (16.5 +/- 0.3 and 14.9 +/- 0.6 mm, P < 0.04). Backfat loss during lactation was positively associated with WOI (P < 0.01) and negatively associated with pregnancy rate (P < 0.04). Plasma leptin concentrations were higher (P < 0.001) in FAT sows than in MEDIUM or THIN sows on both days but there was no relationship between plasma leptin concentrations and reproductive performances after weaning. It is concluded that plasma leptin is associated with backfat depth and that loss of backfat depth during lactation is associated with reproductive performance. However, there is no direct association between plasma leptin and reproductive performance. PMID- 16257600 TI - Effects of in vitro fertilization conditions on preimplantation development and quality of pig embryos. AB - The present study was to investigate the effects of in vitro fertilization conditions on in vitro development and structural integrity of pig embryos. Porcine oocytes matured in vitro were co-incubated with four different spermatozoa concentrations (0.6 x 10(5), 1.2 x 10(5), 2.5 x 10(5) and 5 x 10(5) cells/ml) for 6 h, and at a spermatozoa concentration (1.2 x 10(5) cells/ml) for 2, 4 and 6 h, respectively. Spermatozoa penetration and blastocyst formation were observed at 10 and 144 h post insemination, respectively. The allocation of a blastocyst to inner cell mass (ICM) and trophectoderm (TE) cells was determined by using a differential staining method. Polyspermy frequency increased with increasing spermatozoa concentrations. The spermatozoa-oocyte co-incubation period of 2 h provided for decreased in vitro development rate than 4 and 6 h groups (P < 0.05), although no difference was detected in polyspermy frequency between spermatozoa-oocyte co-incubation periods. Interestingly, blastocysts derived from the groups with greater spermatozoa concentrations (2.5 x 10(5) and 5 x 10(5) cells/ml) had significantly fewer ICM cell nuclei as compared with those groups with lesser spermatozoa concentrations (0.6 x 10(5) and 1.2 x 10(5) cells/ml). There was no difference in the structural integrity of blastocysts among the co-incubation periods. Blastocysts derived from respective experiments were individually classified into three groups (I: <20%; II: 20-40% and III: >40%) based on the ratio of ICM to total cells. Proportion of blastocysts in Group II, with a presumptive normal range of structural integrity, was slightly decreased in the groups with greater spermatozoa concentrations (2.5 x 10(5) and 5 x 10(5) cells/ml). The results indicate that the spermatozoa concentration during in vitro fertilization may be important for developmental competence and quality of pig embryos. PMID- 16257601 TI - Effect of feeding level on progesterone concentration in early pregnant multiparous sows. AB - The effect of three feeding regimens on progesterone level was tested during early pregnancy in multiparous sows. A total of eighteen sows in their eighth parity (8.1 +/- 2.8, mean +/- S.D.) were used. During lactation the sows were fed to appetite and after weaning they received 4 kg (52 MJ) a commercial feed per day. Following ovulation, sows were allocated to one of three treatment groups and fed 2 kg/day (low feeding, LLL) or 4 kg/day (high feeding, HHH) throughout the trial or 2 kg/day for 11 days, 4 kg/day for 10 days, and 2 kg/day for the remaining days of the study (modified feeding, LHL). Blood for progesterone and cortisol analyses was collected daily throughout the study, and for luteinizing hormone (LH) assay for 12 h at 15 min intervals on days 14 and 21 of pregnancy. An adrenocorticotropic hormone (ACTH) challenge test was performed on all sows day 28 of pregnancy. Dietary treatment did not significantly affect hormonal parameters. However, progesterone concentration tended to be lower (P = 0.08) in the HHH group than in the LLL group. In the LHL group venous progesterone concentration seemed to fluctuate. No effects of feeding were observed on progesterone concentration in allantoic fluid on day 35 of pregnancy. Venous cortisol level was significantly higher (P < 0.05) during proestrus and oestrus in all groups and there was no significant difference between groups in response to ACTH challenge. The mean amplitude of LH pulses decreased significantly (P < 0.01) from days 14 to 21 of pregnancy in all groups. In addition, an interaction was found between feeding level and baseline LH concentration and also between feeding level and mean LH concentration. Embryonic recovery was highest in the LLL (69%), lowest in the HHH (45%) and moderate in the LHL (55%) group. Neither high feeding nor modified feeding provided any benefits for reproductive performance in multiparous sows. A low feeding regimen thus appears optimal for multiparous sows in early pregnancy at least with the management regime described. PMID- 16257602 TI - Year-round testicular volume and semen quality evaluations in captive Chinchilla lanigera. AB - In mammals, reproductive performance is usually associated with seasons. Chinchilla lanigera, an endemic South American rodent, reproduces throughout the year in captivity but its seasonal breeding pattern is not fully understood. The present study was designed to evaluate (bi-weekly) over 1 year: (1) testicular volume variations and (2) seminal volume, sperm concentration and functional activity changes. Five animals were studied; they were individually housed indoors (22.2 +/- 1.0 degrees C) under natural photoperiod in Argentina (Cordoba, 31 degrees S-64 degrees W). Semen was obtained by electroejaculation; a total of 116 ejaculates was evaluated. Monthly values for paired testicular volume were less in the middle of the summer than in other seasons (p < 0.006), while those for seminal volume and total spermatozoa/ejaculate were not significantly different; these variables ranged between 7.2-30.9 cm(3), 10-130 microL and 0.9 432.6 x 10(6), respectively. Spermatozoa concentration was (x 10(6)/mL) 2145.9 +/ 365.3 and the pH of semen was 7.3 +/- 0.0. Spermatozoa functional activity showed no significant differences between monthly evaluations; confidence intervals were calculated for the means of: motility, 92.2-95.8%; viability, 92.2 96.1%; swollen cells (hypo-osmotic swelling test), 81.2-87.7% and viable intact acrosome, 83.5-89.0%. The present study represents the first longitudinal reproductive assessment in the chinchilla male. In conclusion, males produce spermatozoa continuously that exhibit high quality functional activity. PMID- 16257603 TI - Age effect of broiler breeders on fertility and sperm penetration of the perivitelline layer of the ovum. AB - The objective of this study was to determine the age effect of a broiler breeder flock on duration of fertility and number of spermatozoa penetrating the perivitelline layer overlying the germinal disc (SP/mm(2) GDIPVL). Moreover, in the second half of the flock's reproductive life, the effect of using ejaculates of young roosters (CA2) in artificial insemination (AI) on the above parameters of fertility was estimated. The commercial flock of broiler breeder hens (n = 100) was inseminated six times from 31 to 62 weeks of age. Additional inseminations, with ejaculates of roosters aged 31 and 36 weeks (CA2), were performed at 56 and 62 weeks of age. AI was performed during two consecutive days (D0 and D1) with an insemination dose of 125 x 10(6) spermatozoa/0.06 ml containing pooled ejaculates. The following parameters were studied: the effective and maximum duration of fertility (De and Dm), percent of fertility on different days after AI (FD10, FD15 and FD20), indices of duration of sperm penetration (DSP, SP < or = 3/GDIPVL), SP/mm(2) GDIPVL in eggs laid on successive days after insemination of hens at different age, and correlations between some fertility indices. Both for De and Dm, the highest values were noted after AI of the layers at 36 weeks of age (14.8 +/- 0.49 and 17.4 +/- 0.46 days, respectively), which were about 2 days longer than at 56 weeks. All fertility indices decreased gradually with age, starting from AI at peak egg production (31 36 weeks of age), while the use of ejaculates from CA2 did not help to increase them significantly. Correlation coefficients between SP/mm(2) GDIPVL and the other fertility indices were positive and highest for eggs laid on D3. It is concluded that high De values can be obtained from broiler breeders in adequate environmental and technological conditions of AI. It is suggested that the age related decrease in fertility is more pronounced in females, in which the efficiency of sperm storage tubules decreases. The present fertility indices indicate the possibility of lengthening AI intervals, especially at peak egg production. PMID- 16257604 TI - Circadian variations in plasma concentrations of melatonin and prolactin during breeding and non-breeding seasons in yak (Poephagus grunniens L.). AB - Circadian variations of plasma melatonin and prolactin concentrations were determined during breeding as well as non-breeding seasons in yak. Blood samples (5 ml) were collected during different phases of estrous cycle, viz. early (0-6 days), mid (7-12 days) and late luteal (13-19 days) at 2 h interval for 24 h from eight yaks during one breeding month (November); the same yaks were bled at 2 h interval during one non-breeding month (February) for 24 h. Plasma melatonin concentrations rose sharply (P < 0.01) after sunset to record peak concentrations between midnight and 2 a.m. declining sharply thereafter in both breeding as well as non-breeding seasons. Basal melatonin concentrations were recorded between 0600 and 1600 h. Stage of luteal phase did not influence the diurnal hormone change (P < 0.01). In the breeding season, mean plasma prolactin concentrations displayed circadian variations with maximum value at 0400 h (41.22+ /- 1.5 ng/ml) and minimum at 1400 h (12.0 +/- 4.02 ng/ml). In the non-breeding season plasma prolactin concentrations showed circadian variation with maximum value at 0000 h (59.9 +/- 10.5 ng/ml) and minimum at 1200 h (32.13 +/- 3.2 ng/ml). A positive correlation in breeding (r = 0.75) and in non-breeding season (r = 0.65) between circadian changes in mean plasma prolactin and melatonin concentrations were seen. Circadian changes of mean plasma melatonin concentrations during breeding and non-breeding seasons were not different (P > 0.05). However, mean plasma prolactin concentrations were found to be higher (P < 0.01) in the non-breeding season. Three conclusions were drawn from the study: (i) melatonin and prolactin concentrations followed a circadian pattern of secretion (ii) melatonin and prolactin secretion may be closely interrelated and (iii) higher prolactin concentrations during the non-breeding season could be due to nutritional and environmental stress and hence might be contributing to lack of cyclicity. PMID- 16257605 TI - Testosterone secretion and pharmacological spermatozoal recovery in the cane toad (Bufo marinus). AB - The cane toad (Bufo marinus) was used as a model to study male anuran reproductive endocrinology and to develop a protocol for non-invasive sperm recovery. Circulating testosterone concentrations in 6-hourly samples did not vary significantly (P < 0.05) over a 24 h period although there was a tendency (P = 0.06) for testosterone to be elevated at 19:00 h relative to other times of the day, which may be related to the nocturnal activity pattern of this species. Testosterone secretion after intraperitoneal (IP) injection of either a GnRH agonist (5 microg IP) or hCG (1000 IU) was also examined. While the GnRH agonist did not produce a significant increase above basal plasma testosterone (0.29, 95% C.I. of 0.05-1.10 ng/ml), injection of hCG resulted in an increase (P < 0.01) of plasma testosterone with peak concentrations at approximately 120 min (4.17, 95% C.I. of 2.69-7.44 ng/ml) after injection. Non-invasive pharmaceutical sperm recovery was attempted following IP injection of graded doses of GnRH agonist, hCG or FSH. Urine was collected at 3, 6 and 12 h after treatment to assess sperm quality and quantity. The optimal protocol for sperm recovery in cane toads was injection of either 1000 or 2000 IU hCG; there was no significant difference in the quality of the spermic urine samples obtained using either dose of hCG or with respect to collection time. The findings indicated that hCG can be used to assess testicular steroidogenic status and also to induce sperm recovery in the cane toad. The hCG protocols developed in this study will have application in studies on the reproductive biology of rare and endangered male anurans. PMID- 16257606 TI - Effect of eCG dose and ovulation induction treatments on embryo recovery and in vitro development post-vitrification in two selected lines of rabbit does. AB - The aim of this work was to evaluate the effect of different doses of eCG administered subcutaneously (0, 50 and 200 IU) and the hormonal induction of ovulation (GnRH or hCG) on embryo recovery and in vitro development of embryos post-vitrification in two selected lines of rabbit does. The two selected lines were line V (selected for the litter size at weaning) and line R (selected for growth rate). Administration of 200 IU of eCG significantly increased ovulation rate (19.2 +/- 1.2 versus 15.5 +/- 1.1 and 12.2 +/- 1.3, and the number of haemorrhagic follicles (13.8+/-1.6 versus 3.8+ /- 1.4 and 3.8 +/- 1.7), but significantly decreased recovery rate (28.8 +/- 6.3 versus 47.7 +/- 5.7 and 48.7 +/- 6.7, 200 IU versus 50 IU and 0 IU eCG, respectively), the number of normal embryos recovered per doe with at least one embryo (5.8 +/- 0.9 versus 8.2 +/- 0.9, 200 IU versus 50 IU eCG doses) and the in vitro development of embryos post vitrification (51.9% versus 66.1%, 200 IU versus 50 IU eCG doses, respectively). Inducing ovulation with hCG significantly increased ovulation rate when compared with GnRH (17.3 +/- 0.8 versus 13.8+/-1.4), but no significant differences in embryo recovery and embryo development post-vitrification were observed between the two treatments. No significant differences were observed between the two selected lines in ovulation and recovery rates, the number of haemorrhagic follicles and the number of recovered embryos per doe. However, the post vitrification in vitro rate of development was 59.7% for line R and 51.9% for line V (p < 0.05). It was concluded that the use of 50 IU of eCG subcutaneous with hCG or GnRH prior to embryo cryopreservation programmes in rabbits achieves the best results for embryo recovery, with the best development of recovered embryos post-vitrification. PMID- 16257607 TI - Experiences in deer sperm cryopreservation under practical conditions--a pilot study. AB - The genetic potential of the red and fallow deer populations in Hungary is well known. Conserving the variability in this excellent genetic material for game preservation is one of our most important task. The aim of the present pilot study was to test the logistical steps of a sperm processing and storing system in which deer sperm can be stored at a level that meets quality standards accepted for domestic animals. Moreover, two different semen extenders, commercially used for freezing bull semen, were compared from the viewpoint of applicability to freeze fallow deer sperm. Sperm was collected from epididymes of eight red stags (Cervus elaphus hippelaphus) and six fallow bucks (Dama dama) during the rutting season. Red deer samples were washed in Triladyl extender, while fallow deer samples were split and processed in Triladyl or Bioxcell extender. In the samples, which had a shorter time interval between the death of the animal and the sperm collection, the percentage of viable spermatozoa with intact acrosome was typically higher. PMID- 16257608 TI - The age of menopause and the menopause transition in a multiracial population: a nation-wide Singapore study. AB - OBJECTIVE: To describe the prevalence of menopausal symptoms, define the mean age of menopause, and determine contributory factors, which influence the experience of symptoms among Singaporean women of different racial groups. DESIGN: Cross sectional nation-wide study of a random sample of 1000 women drawn from the entire population of Singaporean female (Chinese, Malay, and Indian) citizens between 45 and 60 years of age. Face-to-face interviews using structured questionnaires were conducted. RESULTS: The response rate was 69.9%. The mean age of natural menopause was 49.0 years. This was not significantly different between the three ethnic groups. Increasing parity delayed the age of menopause (P=0.007). Muscle and joint ache was the most commonly reported symptom (52.6%). Although the prevalence of significant hot flushes in the general study population was low (3.9%), it was the most commonly reported complaint by peri menopausal women (14.6%). Prevalence of significant hot flushes decreased with time from menopause (P=0.007) and completely disappeared beyond the fifth year of menopause. Recent unhappy events were associated with an increased risk of symptoms (P<0.001). Women of Chinese origin experienced a lower risk of menopausal symptoms when compared with other ethnic groups (P<0.05). CONCLUSION: The mean age of menopause was 49.0 years among Singaporean women. Ethnicity and recent unhappy events were two major factors, which influenced the risk of experiencing menopausal symptoms. Among the three ethnic groups studied, Chinese women were the least likely to experience disturbing menopausal symptoms. The overall prevalence of menopausal symptoms was low when compared to studies on women in western societies. PMID- 16257611 TI - An overview of age of onset of menopause in northern India. AB - OBJECTIVES: This study was conducted to establish the age at onset of natural menopause (ANM) in northern Indian women and also to determine any socio demographic, reproductive or physical factors that may affect it. MATERIALS AND METHODS: Three hundred and fifty women who were accompanying other patients to the outpatient clinic of obstetrics and gynaecology. They came from different parts of northern India since AIIMS is a tertiary health referral centre. All women who had permanent cessation of menstruation for more than 1 year naturally were interviewed (n=201). A semi-structured questionnaire was designed to study the ANM and various demographic, reproductive, and socio-economic factors which affect it. Data were analyzed using logistic regression analysis to see if there was any association between age at natural menopause and various socio demographic factors. RESULTS: The median ANM of the study sample was 48 years with the mean age of 46.7 years. There was no association between the various socio-demographic and reproductive factors and age at onset of menopause. CONCLUSION: The age at onset of menopause in northern India is 48 years which is about 3 years earlier than the west. Socio-demographic and reproductive factors did not affect the age at onset of menopause in our study. PMID- 16257610 TI - The menopause in Spain: results of the DAMES (Decisions At MEnopause) study. AB - OBJECTIVE: To assess the symptomatology of menopause and the use of hormone therapy among women in Spain. METHODS: A survey conducted on a representative sample of 300 women aged 45-55 in Madrid. The instrument included closed- and open-ended questions about demographic and socio-economic information, health and reproductive history, symptoms in past month, use of health services, life style, and therapeutic decisions. RESULTS: Eighty-three percent of respondents report five or more symptoms. Joint pains, weight gain, mood changes and nervousness are the most frequently reported; just under half of respondents report hot flashes; 4/5 report emotional/mental symptoms. The frequencies of some symptoms vary with socio-economic status. Two-thirds of respondents have consulted physicians, and 10% are currently taking hormone therapy; the use of hormone therapy is lower among women from the lower strata. One-fifth of respondents use of nonconventional and alternative medications. Just under half of respondents find that menopause is difficult. CONCLUSION: Symptom reporting is relatively high, but general symptoms are more frequent than "core" menopausal symptoms. The use of hormone therapy is relatively low, but the use of alternative medicines is relatively high. PMID- 16257612 TI - Sun exposure and age at natural menopause: a cross-sectional study in Turkish women. AB - OBJECTIVES: In a cross-sectional study of 157 Turkish women attending outpatient clinics of a university hospital during April-May 2003, association between various subject characteristics and menopause timing was investigated. METHODS: Characteristics were self-reported by women aged 45-60. Of the lifestyle factors, sun exposure, physical activity, food intake and dressing with headscarf were obtained as recalled average lifelong practices up to time of menopause. Cox proportional hazard modeling was used, censoring for hysterectomy, oopherectomy and HRT use. RESULTS: Median age at natural menopause was 52 years. In multivariate analysis, earlier natural menopause was associated with low level of lifelong sun exposure (HR=6.381, 95% CI: 2.996-13.588, p< or =0.0001), heavy physical activity (HR=2.335, 95% CI: 1.305-4.177, p=0.0043), current calcium supplement use (HR=3.191, 95% CI: 1.361-7.485, p=0.0076), diagnosis of hypertension (HR=2.002, 95% CI: 1.186-3.378, p=0.0093), not owning a house (HR=3.002, 95% CI: 1.148-7.852, p=0.0250) and longer years on oral contraceptives (HR=1.085, 95% CI: 1.000-1.176, p=0.0487). Engagement in farming (HR=2.043, 95% CI: 1.056-3.952, p=0.0339), height (cm) (HR=0.953, 95% CI: 0.907-0.994, p=0.0279) and fish consumption (servings/week) (HR=0.600, 95% CI: 0.375-0.960, p=0.0331) were associated with age at menopause in univariate analysis only. For n=109 women who recalled whether maternal menopausal age was <50 or > or =50, sun exposure (HR=7.221, 95% CI: 2.971-17.547, p<0.0001) was a stronger predictor of age at natural menopause than maternal menopausal age (HR=2.882, 95% CI: 1.477 5.621, p=0.0019). CONCLUSIONS: We identify some previously unrecognized correlates of age at natural menopause, namely self-reported lifelong sun exposure, lifelong physical activity, house-ownership, current use of calcium supplements, and lifelong fish consumption. These findings should be confirmed in larger studies. PMID- 16257613 TI - Hormone replacement therapy and hemostasis: effects in Brazilian postmenopausal women. AB - OBJECTIVE: To evaluate the impact that administration of transdermal estradiol gel combined with medroxyprogesterone acetate (MPA) has on hemostasis. METHODS: In this open prospective longitudinal study, thirty postmenopausal women received transdermal estradiol gel (1 mg/day) continuously combined with oral MPA (5 mg/day) for 12 days/month. The following parameters were determined: prothrombin time (PT), activated partial thromboplastin time (aPTT), factors VII, X, and XII activity, fibrinogen levels, thrombin-antithrombin complex levels, protein C and S antigen, antithrombin activity, plasminogen activator inhibitor type 1 (PAI-1) antigen, tissue-type plasminogen activator (t-PA) antigen, plasminogen activity and fibrin degradation products (FbDP) antigen. They were evaluated before and after 6 months of treatment. RESULTS: There was a significant decrease in factor VII activity (P=0.001), factor X activity (P=0.016), protein C antigen (P=0.022), antithrombin activity (P=0.025), plasminogen activity (P=0.023), t-PA antigen (P=0.043) and FbDP antigen (P=0.048) compared with baseline values. CONCLUSION: The results suggest that the treatment with transdermal estradiol gel combined with MPA avoids any major activation of coagulation and does not produce any overall effect on fibrinolysis. Therefore, this treatment might provide interesting effects on hemostasis in postmenopausal Brazilian women. PMID- 16257614 TI - The underdeveloped partnership of patient education and counseling with public health. PMID- 16257615 TI - Informing parents about anaesthesia for children's surgery: a critical literature review. AB - This review critically analysed the published research literature on providing information about children's anaesthesia to parents. An extensive search of the biomedical literature yielded 11 studies: 6 descriptive and 5 intervention studies. Standardised instruments for measuring parental knowledge, need for information and anxiety about children's anaesthetic care showed preliminary validity and reliability. The intervention studies tested different methods of providing information, including verbal, video or written modalities and showed some improvements in knowledge, anxiety and satisfaction. Parents want detailed information about the specifics of anaesthetic procedures, risks, and personnel roles and this information should be incorporated into the routine pre-surgical anaesthesia assessment clinic visits. Until further research determines the most effective methods, mode of delivery should be based on feasibility so that parents consistently receive more information. Audits of parental satisfaction with information should be performed and used to continuously improve the content and delivery of information. PMID- 16257609 TI - Factors associated with climacteric symptoms in women around menopause attending menopause clinics in Italy. AB - OBJECTIVE: To obtain data on correlates of climacteric symptoms in women around menopause attending menopause clinics in Italy. METHODS: Since 1997 a large cross sectional study has been conducted on the characteristics of women around menopause attending a network of first level menopause outpatient's clinics in Italy. A total of 66,501 (mean age 54.4 years) women are considered in the present paper. RESULTS: The odds ratios of moderate and severe hot flashes/night sweats were lower in more educated women and (for severe symptoms only) in women reporting regular physical activity. Depression, difficulty to sleep, forgetfulness and irritability tended to be less frequent in more educated women and (depression only) in women reporting regular physical activity. Parous women reported more frequently these symptoms. CONCLUSIONS: This large study confirms in Southern European population that low education, body mass index and low physical activity are associated with climacteric symptoms. Parous women are at greater risk of psychological symptoms. PMID- 16257616 TI - A cognitive-behavioural program for adolescents with chronic pain-a pilot study. AB - The purpose of this pilot study is to evaluate the feasibility of a cognitive behavioural training program for adolescents with chronic pain irrespective of pain localisation. A secondary aim was to give an impression of the effect of the program on pain and quality of life. Eight adolescents (14-18 years) with chronic non-organic pain recruited from the general population (and their parents) participated in this pilot study. The intervention included five group meetings alternated with four telephone contacts (during the self-management weeks) over a period of 9 weeks. The training aimed to change pain behaviour through pain education, relaxation strategies, problem-solving techniques, assertiveness training, cognitive restructuring and by stimulating the adolescent's physical activity level. The training further addresses the social context of pain by inviting parents to attend two meetings for the parents only, and by asking the adolescents to bring a peer to one of the meetings. Adolescents and their parents were positive about the program. Adolescents felt they were more in control of their pain and parents valued the support they experienced in helping their children to master the pain. The training was considered to be feasible in daily life. Further, the preliminary data showed an effect on pain and quality of life in the expected direction. The results underline the need for a definitive study with a larger sample size and a random controlled design. PMID- 16257617 TI - Patients' perception of preoperative information by interactive computer program exemplified by cholecystectomy. AB - Patients' perception of preoperative information by interactive computer program informing about cholecystectomy was assessed. Patients were asked to qualify the computer program after hospital discharge by an anonymous questionnaire. Two hundred and fifty-seven of 278 consecutive patients completed the questionnaire resulting in a response rate of 92%. Sixty-three percent were very satisfied and 37% were satisfied, whereas none were dissatisfied or undecided. Patients' satisfaction was not associated with sex, age, surgeon or prior knowledge of computers. Most of the patients considered the clarity of text and illustrations and the volume of information as good or excellent and found the information well adapted to their prior knowledge. Ninety-eight percent stated that they would like to be informed again by an interactive computer program in case of another surgical intervention. Thus, interactive computer programs seem to be a valuable adjunct to written and oral information in the preoperative informed consent procedure. PMID- 16257619 TI - The multimedia computer for office-based patient education: a systematic review. AB - Use of the multimedia computer for education is widespread in schools and businesses, and yet computer-assisted patient education is rare. In order to explore the potential use of computer-assisted patient education in the office setting, we performed a systematic review of randomized controlled trials (search date April 2004 using MEDLINE and Cochrane databases). Of the 26 trials identified, outcome measures included clinical indicators (12/26, 46.1%), knowledge retention (12/26, 46.1%), health attitudes (15/26, 57.7%), level of shared decision-making (5/26, 19.2%), health services utilization (4/26, 17.6%), and costs (5/26, 19.2%), respectively. Four trials targeted patients with breast cancer, but the clinical issues were otherwise diverse. Reporting of the testing of randomization (76.9%) and appropriate analysis of main effect variables (70.6%) were more common than reporting of a reliable randomization process (35.3%), blinding of outcomes assessment (17.6%), or sample size definition (29.4%). We concluded that the potential for improving the efficiency of the office through computer-assisted patient education has been demonstrated, but better proof of the impact on clinical outcomes is warranted before this strategy is accepted in the office setting. PMID- 16257618 TI - Asking about intimate partner violence: advice from female survivors to health care providers. AB - Understanding the perspectives of women who have experienced IPV will allow us to identify specific techniques of addressing IPV that increase patient comfort and willingness to disclose and/or seek help. Our study objective was to identify what advice women who had experienced IPV would give health providers regarding how to ask about and discuss the issue of IPV. The women in our study advised that providers (1) give a reason for why they are asking about IPV to reduce women's suspicions and minimize stigma, (2) create an atmosphere of safety and support, (3) provide information, support and access to resources regardless of whether the woman discloses IPV. They emphasized that a provider's asking about IPV is an opportunity to raise patient awareness of IPV, communicate compassion and provide information and not merely a screening test to diagnose a pathologic condition. PMID- 16257620 TI - Striving for quality or length at the end-of-life: attitudes of the Dutch general public. AB - Questionnaires were mailed to 1777 members of the Dutch public (response: 78%), measuring to what extent respondents appreciate life-prolonging treatment, even if it would seriously impair their quality of life. The association between these attitudes and personal characteristics and initiatives to engage in advance care planning was analyzed. About one third of the respondents prefers quality of life at the expense of survival, another third prefers length of life regardless of impaired quality, whereas the remaining third did not express a clear attitude towards quality or length of life. People who were younger, male, having children, having religious beliefs, and without a history of serious illness were more likely to strive for length, whereas the reverse associations were found for striving for quality. The latter was related to undertaking initiatives to engage in advance care planning. Awareness of differences in attitudes towards life prolonging treatment within the public may improve communication about appropriate end-of-life care. PMID- 16257621 TI - Educating families from ethnic minorities in type 1 diabetes-experiences from a Danish intervention study. AB - Ethnic minorities may constitute vulnerable groups within Western health care systems as their ability to master severe chronic diseases could be affected by barriers such as different culture and health/illness beliefs, communication problems and limited educational background. An intervention focusing on immigrant families with children with type 1 diabetes is described. The intervention included the development of adapted educational material and guidelines, and a subsequent re-education of children, adolescents and parents from 37 families. The study demonstrated that it was possible to improve health outcome. During the study, the knowledge of diabetes increased, but with considerable differences between the families. HbA(1c) also decreased significantly during the intervention, but increased during follow-up. The paper discusses possible explanations and suggestions for optimising education and calls for new projects where ethnic minorities are active participants in the development of appropriate educational programs and material. PMID- 16257622 TI - Assessment of mutual understanding of physician patient encounters: development and validation of a Mutual Understanding Scale (MUS) in a multicultural general practice setting. AB - Mutual understanding between physician and patient is essential for good quality of care; however, both parties have different views on health complaints and treatment. This study aimed to develop and validate a measure of mutual understanding (MU) in a multicultural setting. The study included 986 patients from 38 general practices. GPs completed a questionnaire and patients were interviewed after the consultation. To assess mutual understanding the answers from GP and patient to questions about different consultation aspects were compared. An expert panel, using nominal group technique, developed criteria for mutual understanding on consultation aspects and secondly, established a ranking to combine all aspects into an overall consultation judgement. Regarding construct validity, patients' ethnicity, age and language proficiency were the most important predictors for MU. Regarding criterion validity, all GP-related criteria (the GPs perception of his ability to explain to the patient, the patient's ability to explain to the GP, and the patient's understanding of consultation aspects), were well-related to MU. The same can be said of patient's consultation satisfaction and feeling that the GP was considerate. We conclude that the Mutual Understanding Scale is regarded a reliable and valid measure to be used in large-scale quantitative studies. PMID- 16257623 TI - What general practitioners and paediatricians think about their patients' asthma. AB - The Asthma Insight and Reality in Europe (AIRE) study showed that the current management and treatment of asthma in Europe falls short of the goals set in the GINA guidelines. Patient care may be negatively influenced by the physicians' underestimation of their patients' disease state, and overestimation of their patients' knowledge of asthma management. We interviewed 118 paediatricians and 152 general practitioners (response rate 70 and 86%, respectively) in order to get an insight into the physicians view on his patients' asthma management. A questionnaire containing similar items to those used in the AIRE study was used. Dutch physicians believe that the asthma of the majority of their patients is well controlled and underestimate the prevalence of daytime symptoms. They believe that their patients are aware of the differences between reliever medication and maintenance medication and overestimate the number of patients in possession of a written action plan. Dutch paediatricians and general practitioners underestimate the severity of their patients' disease state and overestimate their patients' knowledge of disease management. PMID- 16257624 TI - Can elderly people take their medicine? AB - This study used performance tests to assess the cognitive, visual and physical abilities related to taking medicines in the elderly population. The study population consisted of the Swedish Panel Study of Living Conditions of the Oldest Old (SWEOLD II), a nationally representative interview survey. SWEOLD II is a random sample of all community-based and institutionalized persons aged 77+ in Sweden. Five tests related to medication management were administered in the direct interviews (n=492): hand function (opening bottle), vision (reading label), and medication competence (comprehension and calculation). Results showed that 9.4% could not read instructions on a medicine container and 14.6% had difficulty opening a plastic flip-top medicine bottle. The three cognitive tests related to taking medicine resulted in 30.7, 47.4 and 20.1% errors. Combining all the tests revealed that 66.3% of the sample had at least one limitation of capacity related to taking medicine. There were no significant gender differences. Among those people who did not pass all the tests, 31.8% lived alone with no home-help. Taking medicines is a complex task and a large proportion of the Swedish elderly population has cognitive, visual or physical limitations that may hinder their ability to take medicines accurately. Awareness of these limitations is essential to concordance. PMID- 16257625 TI - Psychological stress as a predictor of psychological adjustment and health status in patients with rheumatoid arthritis. AB - This study examined the extent to which perceived stress, social support, coping and clinical disease indicators predict physical, psychological and social adjustment in patients with rheumatoid arthritis (RA). Participants were 59 women recruited at an outpatient clinic at University College Hospital, Galway. A range of psychological measures was administered and disease status was assessed by physician ratings of joint involvement and blood assays of inflammatory indices. Findings from correlational and hierarchical regression analyses revealed a number of statistically significant relationships (p<.01). Perceived stress was a better predictor than disease severity of positive and negative emotionality. Coping explained variability on positive and negative affect. Social support was linked to level of social activity. Results demonstrated that disease status predicted illness related functioning but did not predict emotional or social adjustment. Results suggest that a cognitive behavioural intervention to facilitate patient adjustment could usefully include management of stress and its appraisal, the fostering of adaptive coping strategies and utilization of social support resources. It is concluded that improving patient adjustment to rheumatoid arthritis has implications for medical care seeking. PMID- 16257626 TI - Development of an interactive computer-assisted instruction (ICAI) program for patient prenatal genetic screening and carrier testing for use in clinical settings. AB - Educating patients on prenatal genetic screening and carrier testing in a timely and effective manner is faced with barriers including, providers' limited knowledge, and little time available to spend discussing screening and testing during a visit. This paper describes the use of cognitive response interviews (CRI) and usability testing (UT) in the development of an interactive computer assisted instruction (ICAI) program for use by prenatal patients in clinical settings. Lessons learned during the program development process included simplification of content and adaptation of navigational features in response to observations and interviews of a sample of patients representing the intended population. The resulting program functions as a targeted patient education program that maintains the level of medical information needed, as specified by professional practices guidelines, in a patient friendly format. In addition, this ICAI program functions as a research tool that can collect data on program effectiveness. Researchers developing other patient education programs will benefit from the lessons learned during development of this ICAI program by considering rephrasing of content to fit patient understanding, and adding navigational features to help further facilitate effective program use. PMID- 16257629 TI - Phenotype definition, age, and gender in the genetics of asthma and atopy. AB - When studying genetics of complex diseases it is important to have a clearly described and objective phenotype. When drawing conclusions in association studies, age and gender of the population should be considered. Until we know what causes phenotypic differences between males and females and between children and adults, we should try to study longitudinal cohorts with phenotype assessment at different time points and stratify our analyses for gender. To acquire sufficient power for these types of analyses, international collaboration may be the only way to elucidate the intricate, gene-environmental interactions in atopy and asthma in an age- and gender-dependent manor. PMID- 16257630 TI - Family studies and positional cloning of genes for asthma and related phenotypes. AB - Although it is not yet known how many genes may contribute to the susceptibility or the severity of asthma and related phenotypes, genome-wide screens and positional cloning techniques have been successful in identifying contributing genes in multiple populations. The results of these studies provide additional insight into the molecular mechanisms responsible for the development of a variety of phenotypes. Replication with additional populations--particularly in large-scale studies--has been used to distinguish between false positive results or population-specific effects or to further quantify the conferred risk. Even when individual markers do not replicate in multiple population, association of the same region or gene has been useful in directing future studies. As further understanding of linkage disequilibrium patterns within the genome has allowed greater efficiency for genetic studies, advances in high-throughput genotyping technology, genetic analysis methodologies, and a more in-depth understanding of clinical phenotypes has made genome-wide studies more accessible and cost effective. In the future, identification of function variants with clinical relevance may be used to influence the diagnosis and treatment of asthma. PMID- 16257627 TI - Effects of distance learning on clinical management of LUTS in primary care: a randomised trial. AB - AIM: To determine the effect of a distance learning programme on general practice management of men with lower urinary tract symptoms (LUTS). METHODS: A cluster randomised controlled trial was performed. General practitioners (GPs) were randomised to a distance learning programme accompanied with educational materials or to a control group only receiving mailed clinical guidelines on LUTS. Clinical management was considered as outcome. RESULTS: Sixty-three GPs registered care management of 187 patients older than 50 years attending the practice because of LUTS. The intervention group showed a lower referral rate to a urologist (OR: 0.08 (95% CI: 0.02-0.40)), but no effect on PSA testing or prescription of medication. PSA testing tended to be requested more frequently by intervention group GPs. Secondary analysis showed patients in the intervention group received more educational materials (OR: 75.6 (95% CI: 13.60-419.90)). CONCLUSIONS: The educational programme had impact on clinical management without changing PSA testing. Distance learning is an promising method for continuing education. PRACTICE IMPLICATIONS: Activating distance learning packages are a potentially effective method for improving professional performance. Emotional matters as PSA testing probably need a more complex approach. PMID- 16257631 TI - ADAM33: a newly identified gene in the pathogenesis of asthma. AB - There is much to find out about this fascinating and complex molecule in relation to the development and progression of asthma. Added to it are three further new asthma/allergy genes identified by positional cloning: PDH Finger Protein II (PHF11) on chromosome 13q14, which encodes NY-REN-34 a protein first described in patients with renal cell carcinoma [67]; Dipeptidyl diptidase 10 (DDP10) on chromosome 2q14 [68]; and G protein-coupled receptor for asthma susceptibility (GPRA) on chromosome 7p [69]. For each of these genes, as is the case for ADAM33, determining their normal function(s) and how these become disordered in asthma is the future challenge. PMID- 16257632 TI - HLA-G: an asthma gene on chromosome 6p. AB - We identified HLA-G as an asthma susceptibility gene in multiple populations and demonstrated that variation in this gene influences subsequent risk for asthma. Prenatal exposure to factors that are correlated with maternal BHR (or perhaps BHR itself) interacts with fetal genotype to determine risk, however. Among fetuses of unaffected mothers, the +1489TT genotype is a marker for increased risk, whereas among fetuses of affected mothers the +1489CC genotype is a marker for increased risk. Studies are underway to understand the mechanism for this interaction and the role of this gene in the pathogenesis of asthma. PMID- 16257633 TI - Candidate gene association studies and evidence for gene-by-gene interactions. AB - Candidate gene studies in asthma are a powerful and valuable tool in asthma genetics. Although the quality of small-scale, freely associating studies has been questionable, increasingly serious efforts are made to establish, replicate, and verify association results. Association studies may help us to better understand the mechanisms underlying asthma. They may create hypotheses and help to direct functional studies to targets that are likely to give valuable results. However, they should not be over-interpreted; only biologic proof can verify associations between genetic variations and a certain disease outcome. The insight that gene-by-gene and gene-by-environment interactions may be crucial for understanding and pinpoint the complex mechanisms of genetic regulation of multifactorial diseases has gained momentum in the last years when technical improvement allowed for the effective genotyping and analysis of great numbers of polymorphisms in large populations. It can be expected that from this area of research new and exciting results will follow soon. PMID- 16257634 TI - Gene-environment interactions in asthma and allergies: a new paradigm to understand disease causation. AB - The example of complex interactions between environmental exposures and polymorphisms in the CD14 gene in predisposing for allergy-related conditions offers a good indication of the complexity of the mechanisms that determine susceptibility to these conditions. Contrary to what has been the rule for monogenic diseases, the association between genetic variations and polygenic conditions such as asthma and allergies may not always be unidirectional; that is, not always will the same alleles be associated with the conditions under study. Concepts of penetrance of genetic variations that ignore these nonlinear influences (which may affect gene-gene and gene-environment interactions) may hinder a better understanding of mechanisms of disease involved, and therefore may delay the development of preventive strategies for these common conditions. Discrepancies between well-designed genetic studies of asthma and allergies, therefore, may be suggesting something fundamental about how these diseases develop and how it will be possible to abolish them in the future. PMID- 16257635 TI - Asthma pharmacogenomics. AB - It is expected that future treatments will be preceded by genetic tests to prescribe the most effect asthma medication while lowering the risk of adverse side effects. However, it will not be necessary to describe all the genetic determinants affecting drug response to apply pharmacogenomics to asthma therapy. Whether pharmacogenomics becomes common practice may not depend on the availability of tests, but on factors such as affordability, ease of application, and ease of interpreting the results. PMID- 16257636 TI - New approaches to understanding the genetics of asthma. AB - Several key conditions that are necessary to identify disease susceptibility genes in common diseases such as asthma are now available, including (1) increasingly comprehensive genomic information on gene location, genomic structure, and sequence variants, from the Human Genome Project (and from other species); (2) better understanding of the biologic functions of relevant genes and inflammatory and immunity pathways important in asthma; (3) newer high throughput and accurate technologies for DNA sequencing and SNP genotyping; (4) improved statistical methods for analyzing genetic data from families and populations; and (5) availability of methods to characterize function of sequence variants and study biologic responses. Collectively, these conditions will allow the prioritization of candidate genes based on available knowledge of map position and biologic relevance; obtain genomic structure of these genes; and study sequence variants in these genes in populations to facilitate the identification of genes that are important in the development and expression (severity) of asthma and associated phenotypes. Although, it is still a labor intensive and expensive project to identify susceptibility genes in common diseases such as asthma, the new techniques that are now being used will greatly facilitate gene mapping. The techniques discussed in this article include genome by-genome analysis in family data, such as those listed in Box 2. This analysis has already been shown to be a powerful too in mapping genes for another common disease (prostate cancer) with interesting preliminary results for asthma. Second, the use of man-mouse homology mapping that has proven very useful in cardiovascular studies is beginning to be applied to asthma and related phenotypes. finally with new available technology, genome-wide screens using very dense SNP maps are now a reality and a significant new development in family linkage and case-control association studies. In summary, these new approaches should be considered in designing studies to detect genes that are important in asthma and allergy. PMID- 16257637 TI - The evolution of Free Radical Biology & Medicine: a 20-year history. PMID- 16257638 TI - Mitochondrial respiratory chain and free radical generation in stroke. AB - Being the second most common cause of death in the industrial countries and one of the major causes of death and disability, stroke has a great effect on public health and is the neurological disease which accounts for the largest number of hospitalizations. In order to develop new treatments, biochemical mechanisms involved in brain damage have been investigated. Among them, oxidant species generated during stroke have been implicated as critical mediators of neuronal injury in this condition, although neuroprotective roles have also been demonstrated. This review is focused on the role of the mitochondrial respiratory chain as both source and target of reactive oxygen and nitrogen species such as nitric oxide, superoxide and peroxynitrite produced in cerebral ischemia. The neuroprotective role of antioxidants or other molecules acting on the mitochondrial respiratory chain and ATP synthesis in the setting of cerebral ischemia is discussed. PMID- 16257639 TI - Iron and oxidative injury-- a commentary on "Fatty acid-mediated iron translocation: a synergistic mechanism of oxidative injury" by D. Yao et al. PMID- 16257640 TI - Delivery of orally supplemented alpha-tocotrienol to vital organs of rats and tocopherol-transport protein deficient mice. AB - The natural vitamin E tocotrienol (TCT) possesses biological properties not shared by tocopherols (TCP). Nanomolar alpha-TCT, not alpha-TCP, is potently neuroprotective (JBC 275:13049; 278:43508). Tocopherol-transport protein (TTP) represents the primary mechanism for maintaining normal alpha-TCP concentrations in plasma and extrahepatic tissues. TTP primarily transports alpha-TCP and has low affinity for alpha-TCT. There are no studies that have investigated tissue delivery of alpha-TCT when orally gavaged on a long-term basis. A long-term study was conducted to examine the effects of alpha-TCT or alpha-TCP supplementation, either alone or in combination, on tissue levels. Rats were maintained on a vitamin E-deficient diet and gavaged with alpha-TCT or alpha-TCP alone or in combination. Five generations of rats were studied over 60 weeks. TTP-deficient mice were supplemented with TCT and bred to examine tissue delivery of oral alpha TCT. Orally supplemented alpha-TCT was effectively delivered to most tissues over time. When co-supplemented, alpha-TCP outcompeted alpha-TCT for transport systems delivering vitamin E to tissues. To evaluate the significance of TTP in alpha-TCT delivery to tissues, tissue levels of alpha-TCT in supplemented TTP-deficient mice were studied. alpha-TCT was transported to several vital organs in TTP deficient mice. alpha-TCT restored fertility in TTP-deficient mice. In sum, orally supplemented alpha-TCT was successfully delivered to several vital organs. The transport efficiency of alpha-TCT to tissues may be maximized by eliminating the co-presence of alpha-TCP in the oral supplement. Examination of whether alpha TCT may benefit humans suffering from neurological disorders because of congenital TTP deficiency is warranted. PMID- 16257641 TI - Oxidative stress disrupts nitric oxide synthase activation in liver endothelial cells. AB - Oxidative stress may mediate vascular disruption associated with a loss of endothelial nitric oxide synthase (eNOS) activity and a hypersensitivity to the constrictor effects of endothelin-1 (ET-1). We hypothesize that this is due, in part, to uncoupling of ET(B) receptors from eNOS activation. Thus, we tested whether oxidative stress (OS) affects liver vascular relaxation by reducing basal and ET-1-induced NO production. Primary sinusoidal endothelial cell cultures were pretreated with H(2)O(2) (25 microM) for 1 or 6 h before a 10-min ET-1 stimulation. OS resulted in a significant basal and ET-1-induced decrease in NO production. Acute OS increased the monomeric form of the inhibitory protein caveolin-1 (1.2 +/- 0.05 vs 0.9 +/- 0.02, p < 0.01) and increased the eNOS caveolin association as determined by coimmunoprecipitation (1.24 +/- 0.04 vs 0.97 +/- 0.04, p < 0.05). ET-1 stimulation further exacerbated these effects. Subacute OS inhibited ET-1-induced eNOS phosphorylation of serine 1177 (activation residue) (1 +/- 0.07 vs 1.6 +/- 0.04, p < 0.05) and dephosphorylation of the inhibitory residue threonine 495 (1.5 +/- 0.08 vs 0.7 +/- 0.02, p < 0.01). Additionally subacute OS resulted in dissociation of eNOS from ET(B) (0.8 +/- 0.09 vs 1.2 +/- 0.06, p < 0.05). Our findings indicate that acute and subacute oxidative stress result in the inhibition of induced nitric oxide synthase activity through distinct mechanisms dependent on caveolin-1 inhibition, ET(B) dissociation, and eNOS phosphorylation. PMID- 16257642 TI - Overexpression of MsrA protects WI-38 SV40 human fibroblasts against H2O2 mediated oxidative stress. AB - Proteins are modified by reactive oxygen species, and oxidation of specific amino acid residues can impair their biological functions, leading to an alteration in cellular homeostasis. Oxidized proteins can be eliminated through either degradation or repair. Repair is limited to the reversion of a few modifications such as the reduction of methionine oxidation by the methionine sulfoxide reductase (Msr) system. However, accumulation of oxidized proteins occurs during aging, replicative senescence, or neurological disorders or after an oxidative stress, while Msr activity is impaired. In order to more precisely analyze the relationship between oxidative stress, protein oxidative damage, and MsrA, we stably overexpressed MsrA full-length cDNA in SV40 T antigen-immortalized WI-38 human fibroblasts. We report here that MsrA-overexpressing cells are more resistant than control cells to hydrogen peroxide-induced oxidative stress, but not to ultraviolet A irradiation. This MsrA-mediated resistance is accompanied by a decrease in intracellular reactive oxygen species and is partially abolished when cells are cultivated at suboptimal concentration of methionine. These results indicate that MsrA may play an important role in cellular defenses against oxidative stress, by catalytic removal of oxidant through the reduction of methionine sulfoxide, and in protection against death by limiting, at least in part, the accumulation of oxidative damage to proteins. PMID- 16257643 TI - Coronary microvascular endothelial cell growth regulates expression of the gene encoding p22-phox. AB - The mRNA expression of p22-phox, a membrane-bound component of NAD(P)H oxidase, is elevated in proliferating compared to quiescent rat coronary microvascular endothelial cells (CMEC). However, the underlying molecular mechanisms of this increase and its relevance to cell confluence remain unknown. This study shows greater levels of p22-phox mRNA/protein expression, NAD(P)H oxidase activity, and superoxide anion (O(2)(-)) production in proliferating versus fully confluent and growth-arrested 50% confluent CMEC. Both enhanced transcription of the p22-phox gene and enhanced mRNA half-life were shown to contribute to the increase in p22 phox mRNA levels as demonstrated by nuclear run-on studies and Northern analyses after actinomycin D transcriptional arrest, respectively. Suppression of O(2)(-) availability by a cell-permeable superoxide dismutase mimetic or inhibition of NAD(P)H oxidase activity via its specific inhibitors (apocynin, diphenyleneiodinium, phenylarsine oxide, and 4-(2-aminoethyl)bezenesulfonyl fluoride) or antisense p22-phox cDNA attenuated cell growth independent of changes in intracellular antioxidant glutathione levels. Inhibition of nascent protein synthesis by a translation inhibitor, cycloheximide, significantly reduced p22-phox mRNA stability in proliferating but not in confluent CMEC. These findings suggest that expression of p22-phox mRNA in CMEC is upregulated by growth through a mechanism that involves increases in transcriptional activity of the p22-phox gene and stability of the p22-phox message after its formation. PMID- 16257644 TI - Peroxynitrite increases VEGF expression in vascular endothelial cells via STAT3. AB - Increased expression of vascular endothelial growth factor (VEGF) has been correlated with increased oxidative stress and formation of peroxynitrite in numerous disease conditions, including diabetic microangiopathy, tumor angiogenesis, and atherosclerosis. In this study we tested the hypothesis that peroxynitrite stimulates VEGF expression. Treatment of microvascular endothelial cells with exogenous peroxynitrite induced a time- and dose-dependent increase in VEGF mRNA, which peaked within 1 h of treatment at a concentration of 100 muM. The increase in VEGF mRNA was followed by a significant increase in VEGF protein. To define the molecular mechanisms involved, the effect of peroxynitrite was determined on the activation of two transcription factors known to regulate VEGF expression during hypoxia and tumor angiogenesis-signal transducer and activator of transcription 3 (STAT3) and hypoxia-inducible factor-1 (HIF-1). Peroxynitrite caused activation and nuclear translocation of STAT3, but not HIF-1. Moreover, transduction of endothelial cells with dominant-negative STAT3 abrogated the peroxynitrite-induced increase in VEGF mRNA. The increase in VEGF mRNA was also blocked by inhibitors of transcription and was unaffected by the inhibition of protein synthesis. These results indicate that peroxynitrite causes increased expression of VEGF in vascular endothelial cells by a process that requires the activation of STAT3. PMID- 16257645 TI - High sensitivity enzyme-linked immunosorbent assay (ELISA) method for measuring protein carbonyl in samples with low amounts of protein. AB - The oxidative modification of proteins has been shown to play a major role in a number of pathological processes. One such modification is the addition of the carbonyl groups to the amino acid residue in proteins. For the measurement of the carbonyl groups in low concentration protein samples, we have modified the ELISA (enzyme-linked immunosorbent assay) method that was developed by Buss et al. [Buss, I. H; Chan, T. P.; Sluis, K. B.; Domigan, N. M.; Winterbourn, C. C. Protein carbonyl measurement by a sensitive ELISA method. Free Radic. Biol. Med.23:361-366; 1997 ]. In the modified method, protein samples diluted in phosphate-buffered saline were adsorbed to wells of an ELISA plate and then reacted with dinitrophenylhydrazine (DNPH). The protein-conjugated DNPH was probed by a commercial anti-DNPH antibody, and then a second antibody conjugated with horseradish peroxidase was added for quantification. The method was calibrated using oxidized albumin, and required only 5 mug protein. This obviated the need to concentrate protein in experimental and clinical samples with low amounts of protein. In addition the effect of TCA on carbonyl measurement is eliminated. The standard curve was linear in the range of 0-3.36 nmol carbonyls/mg protein, which is the range within which clinical samples fell. The results correlated well with the colorimetric carbonyl assay. The method was used to analyze the amount of protein carbonyl in aqueous humor and diluted plasma samples. PMID- 16257646 TI - Cytotoxicity and cytoprotective activity in naphthalenediols depends on their tendency to form naphthoquinones. AB - We consider the cytotoxicity and the protection against oxidative stress for members of the naphthalenediol family and the known antioxidant epigallocatechin gallate (EGCG). Compounds include the 1,2-naphthalenediol (1,2-ND), 1,4-ND, 2,3 ND, 1,8-ND, and 1,4-dipropyl-2,3-naphthalenediol (DPND). The cell line is an adherent clone of rat pheochromocytoma (PC12-AC). Oxidative stress was induced by the peroxyl radical generator AAPH. The relative order of cytotoxicity was 1,4-ND > 1,2-ND > DPND > 2,3-ND > 1,8-ND > EGCG, with EC(50)'s of 15, 40, 160, >250, >250, >>250 muM, respectively. Despite their high toxicity, both 1,4-ND and 1,2 ND showed narrow zones of protective behavior whereas DPND, 2,3-ND and 1,8-ND and especially EGCG showed an extended protective range. The total protection obtained for the combination of cells/oxidative stressor/protective compounds (PC12-AC/AAPH/naphthalenediols) was defined by an integrated measure, the cytoprotective area (CPA). We relate the observed cytotoxicity and CPA to the different electronic structures of the naphthalenediols, characterized by the first and second bond dissociation enthalpies and the pK(a)'s for parent (diol) and semiquinone. Since the 2,3- and 1,8-naphthalenediols do not form quinones, their cytotoxicity is much lower than for the compounds which do. Thus selected members of the naphthalenediol family show promise as antioxidants. PMID- 16257647 TI - Cadmium chloride-induced oxidative stress in skeletal muscle cells in vitro. AB - The effects of cadmium chloride (CdCl(2)) on oxidative stress in the skeletal muscle cell line C(2)C(12) were investigated. Myoblast cells that differentiated into myotubes were treated with CdCl(2) (1, 3, 5, 7.5, 10, and 12.5 microM) for 24, 48, and 72 h. Subsequent assay of cell homogenates for MTT (3-(4,5 dimethylthiozol-2-yl)-2,5-diphenyltetrazolium bromide) reduction, neutral red uptake and nucleic acid content showed that cadmium was toxic to C(2)C(12) cells in a concentration-dependent manner. Glutathione-S-transferase activity (nmol microg of protein(-1) min(-1)) was increased with 1 and 3 microM CdCl(2) (36.9 +/ 5.6 and 32.1 +/- 6.0, respectively) compared to control cells (21.8 +/- 1.5), but decreased at higher concentrations (7.5 microM = 15.9 +/- 3.3, 10 microM = 15.9 +/- 4.6, and 12.5 microM = 10.5 +/- 2.8). An increase in malondialdehyde content (nmol microg of protein(-1)), especially at high CdCl(2) concentrations (control = 7.3 +/- 0.5; CdCl(2): 7.5 microM = 11.2 +/- 3.1, 10 microM = 14.6 +/- 3.8, and 12.5 microM = 20.5 +/- 6.5) indicated that there was enhanced lipid peroxidation. Light and scanning electron microscopy showed that there was a concentration-dependent loss of adherent cells and the formation of vesicles indicative of cell death. These results indicated that CdCl(2) increased oxidative stress in C(2)C(12) cells, and this stress probably compromised cell adhesion and the cellular antioxidant defense mechanisms. PMID- 16257648 TI - Fatty acid-mediated intracellular iron translocation: a synergistic mechanism of oxidative injury. AB - Fatty acid has been reported to be associated with cardiovascular diseases and cancer, but the possible mechanism remains unclear. Here, we reported a novel mechanism for the permissive role of fatty acid on iron intracellular translocation and subsequent oxidative injury. In vitro study from endothelial cells showed that iron alone had little effect, whereas in combination with PA (palmitic acid), iron-mediated toxicity was markedly potentiated, as reflected in mitochondrial dysfunction, cell death, apoptosis, and DNA mutation. We also showed that PA not only facilitated iron translocation into cells through a transferrin-receptor (TfR)-independent mechanism, but also translocated iron into mitochondria; the subsequent intracellular iron overload resulted in reactive oxygen species (ROS) overgeneration and lipid oxidation. Further investigation revealed that PA-facilitated iron translocation is due to Fe/PA-mediated extracellular oxidative stress and the subsequent membrane damage with increased membrane permeability. Fe/PA-mediated toxic effects were reduced in rho0 cells lacking mitochondrial DNA or by antioxidant enzyme SOD, especially mitochondrially localized MnSOD, suggesting a permissive role of PA for iron deposition on the vascular wall and its subsequent toxicity via mitochondrial oxidative stress. This observation was confirmed in vivo in mice, wherein higher vascular iron deposition and accompanying superoxide release were observed in the presence of a high-fat diet with iron administration. PMID- 16257649 TI - Pulse radiolysis study of the interaction of retinoids with peroxyl radicals. AB - Vitamin A (retinol) and its derivatives-retinal and retinoic acid-are known for their ability to inhibit lipid peroxidation. Antioxidant actions of retinoids have been attributed to chain-breaking by scavenging of peroxyl radicals. Based on chemical analysis of retinoic acid degradation products formed during microsomal lipid peroxidation, it was previously suggested that retinoids interact with peroxyl radicals forming free carbon-centered radical adducts. However, it can be argued that such a mode of antioxidant action of retinoids is not sufficient to fully explain their effectiveness at inhibiting lipid peroxidation, which in many systems is comparable to, or even exceeds, that of alpha-tocopherol. In order to elucidate the mechanism of interaction of retinoids with peroxyl radicals, (trichloromethyl)peroxyl radical was generated by pulse radiolysis, and its interactions with retinoids solubilized in Triton X-100 micelles were followed by kinetic absorption spectroscopy. All retinoids- retinol, retinal, and retinoic acid--interacted with the peroxyl radical, and at least two transient products were detected. One of these products, absorbing at 590 nm, was identified as retinoid cation radical. Therefore, we postulate that, apart from formation of radical adducts, retinoids may also scavenge peroxyl radicals by electron transfer. PMID- 16257651 TI - A QCM immunosensor for Salmonella detection with simultaneous measurements of resonant frequency and motional resistance. AB - A quartz crystal microbalance (QCM) immunosensor was described for the detection of Salmonella Typhimurium with simultaneous measurements of the resonant frequency and motional resistance. The immunosensor was fabricated using protein A for the antibody immobilization. High-frequency impedance analysis indicated that the changes in resonant frequency and motional resistance (DeltaF and DeltaR) of the QCM were significant while the changes in static capacitance, motional capacitance, and motional inductance were insignificant. In the direct detection of S. Typhimurium in chicken meat sample, DeltaF and DeltaR were proportional to the cell concentration in the range of 10(5) - 10(8) and 10(6) - 10(8) cells/ml, respectively. Using anti-Salmonella-magnetic beads as a separator/concentrator for sample pretreatment as well as a marker for signal amplification, the detection limit was lowered to 10(2) cells/ml based on the DeltaR measurement; however, DeltaF was not related to the cell concentration. No interference was observed from E. coli K12 or the sample matrix. PMID- 16257652 TI - Micromechanical cantilever array sensors for selective fungal immobilization and fast growth detection. AB - We demonstrate the use of micromechanical cantilever arrays for selective immobilization and fast quantitative detection of vital fungal spores. Micro fabricated uncoated as well as gold-coated silicon cantilevers were functionalized with concanavalin A, fibronectin or immunoglobulin G. In our experiments two major morphological fungal forms were used--the mycelial form Aspergillus niger and the unicellular yeast form Saccharomyces cerevisiae, as models to explore a new method for growth detection of eukaryotic organisms using cantilever arrays. We exploited the specific biomolecular interactions of surface grafted proteins with the molecular structures on the fungal cell surface. It was found that these proteins have different affinities and efficiencies to bind the spores. Maximum spore immobilization, germination and mycelium growth was observed on the immunoglobulin G functionalized cantilever surfaces. We show that spore immobilization and germination of the mycelial fungus A. niger and yeast S. cerevisiae led to shifts in resonance frequency within a few hours as measured by dynamically operated cantilever arrays, whereas conventional techniques would require several days. The biosensor could detect the target fungi in a range of 10(3) - 10(6) CFUml(-1). The measured shift is proportional to the mass of single fungal spores and can be used to evaluate spore contamination levels. Applications lie in the field of medical and agricultural diagnostics, food- and water-quality monitoring. PMID- 16257653 TI - Electronic nose screening of ethanol release during sol-gel encapsulation. A novel non-invasive method to test silica polymerisation. AB - Porous silica matrices prepared by sol-gel process yield biocompatible materials adequate for encapsulation of biomolecules or drugs. The procedure is simple and fast, but when alkoxyde precursors like tetraethoxysilane (TEOS) are used the polymerisation reaction leads to the formation of alcohol as a by-product, which can produce undesirable effects on the activity of entrapped enzymes or modify a drug release kinetic. Therefore, it is critical to determine that no remnant ethanol is left prior using or storing the obtained biomaterial. In this regard, the technique used in the alcohol determination should be non-invasive and non destructive to preserve the encapsulation device intact and ready to use. In this work we have successfully used a portable electronic nose (e-nose) for the screening of silica polymerisation process during theophylline encapsulation. TEOS reaction was "smelt" since precursor pre-hydrolysis until the end of ethanol release, sensed directly at the headspace of matrices slabs. Measurements showed that ethanol was negligible since 10th day in polymeric slabs of 10 mm width and 2 cm diameter. This first use of e-nose following a polymerisation reaction opens a wide number of putative applications in pharmaceutical and biochemical fields. PMID- 16257654 TI - A novel electrochemical detection method for aptamer biosensors. AB - A beacon aptamer-based biosensor for the detection of thrombin was developed using electrochemical transduction method. Gold surface was modified with a beacon aptamer covalently linked at 5'-terminus with a linker containing a primary aliphatic amine. Methylene blue (MB) was intercalated into the beacon sequence, and used as an electrochemical marker. When the beacon aptamer immobilized on gold surface encounters thrombin, the hairpin forming beacon aptamer is conformationally changed to release the intercalated MB, resulting a decrease in electrical current intensity in voltamogram. The peak signal of the MB is clearly decreased by the binding of thrombin onto the beacon aptamer. The linear range of the signal was observed between 0 and 50.8 nM of thrombin with 0.999 correlation factor. This method was able to linearly and selectively detect thrombin with a detection limit of 11 nM. PMID- 16257656 TI - Rapid PCR confirmation of E. coli O157:H7 after evanescent wave fiber optic biosensor detection. AB - Escherichia coli O157:H7 is an enteric pathogen of public health importance, which is monitored by several government agencies. Many rapid detection tests have been developed to identify foodstuff and water supplies contaminated by E. coli O157:H7. However, these methods can be time consuming (24-48 h) due to the need to culture the bacteria to confirm detection results. Fiber optic biosensors can rapidly detect pathogens from complex matrices, yet confirmation tests can take up to 10h to complete. In addition, fiber optic biosensors can also be used to reduce the impact of PCR inhibitors present in complex matrices such as food and water. This paper presents methodologies to reduce the time necessary for confirmation from 10 to about 2 h, by direct PCR of bacteria from the fiber optic waveguides without the need for culture or enrichment steps. PMID- 16257655 TI - Detection of pathogen Escherichia coli O157:H7 AT 70 cells/mL using antibody immobilized biconical tapered fiber sensors. AB - Optical fibers (core diameter 8 microm, cladding diameter 125 microm) was tapered to a waist diameter in the range of 8-12 microm, and then a monoclonal antibody to the pathogen, Escherichia coli O157:H7 was covalently bonded to the surface of the tapered region. Using 470 nm light, the taper was exposed to various concentrations (7 x 10(7), 7 x 10(5), 7 x 10(3), and 70 cells/mL) of the pathogen, and the sensor showed changes in transmitted light as the antigen attached to the antibody on the taper surface. The response was equal and opposite when the pathogen was released from the surface using a low pH buffer. The magnitude of the change was inversely proportional to the concentration of the pathogen. The sensor showed good sensitivity at as low a concentration as 70 cells/mL. The antibody-immobilized taper sensor was also exposed to a mixture of the pathogen and a non-pathogenic variant (JM101) at 0%, 50% and 70% by concentration. The sensor showed good selectivity to the pathogenic antigen. A first order attachment kinetic model is proposed to quantify the rate of attachment of pathogen to the sensor surface. The kinetic rate constant (k) of E. coli O157:H7 to the fiber was found to vary in the range of (2.5-6.1) x 10(-9) min(-1) (cells/mL)(-1). PMID- 16257657 TI - Single base mismatch detection by microsecond voltage pulses. AB - A single square voltage pulse applied to metal electrodes underneath a silicon dioxide film upon which DNA probes are immobilized allows the discrimination of DNA targets with a single base mismatch during hybridization. Pulse duration, magnitude and slew rate of the voltage pulse are all key factors controlling the rates of electric field assisted hybridization. Although pulses with 1 V, lasting less than 1 ms and with a rise/fall times of 4.5 ns led to maximum hybridization of fully complementary strands, lack of stringency did not allow the discrimination of single base mismatches. However, by choosing pulse conditions that are slightly off the optimum, the selectivity for discriminating single base mismatches could be improved up to a factor approximately 5 when the mismatch was in the middle of the strand and up to approximately 1.5 when the mismatch was on the 5'-end and. These results demonstrate that hybridization with the appropriate electric field pulse provides a new, site-specific, approach to the discrimination of single nucleotide polymorphisms in the sub-millisecond time scale, for addressable DNA microarrays. PMID- 16257658 TI - Surface plamon resonance imaging of DNA based biosensors for potential applications in food analysis. AB - The adsorption processes of oligonucleotides immobilised onto suitable photolithographic patterned gold substrates have been investigated in aqueous buffer solution by using a home made surface plasmon resonance (SPR) imaging equipment. A rapid self-assembled method for the construction of DNA chips to be used in SPR imaging experiments have been followed. The immobilised DNA molecules (probes) adopted in our SPR experiments anchored to a gold surface via thiol group were 5'thiol-modified containing a (CH(2))(15) tail. The hybridisation processes taking place with its complementary sequence have been observed and characterized by monitoring phenomena by a SPR imaging system. The two analysed oligonucleotides (probes and target) are of interest in plant gene biotechnological application and differing for the presence at the 5'-end of a poly T16 spacer. Dynamic investigation of smallest changes in SPR imaging pictures performed in liquid phase in the presence of DNA complementary probes have been performed. Quantitative information in terms of threshold of sensitivity has been extracted by using a specific images treatment. PMID- 16257659 TI - Size-selective recognition of catecholamines by molecular imprinting on silica alumina gel. AB - The preparation of a catecholamine receptor was carried out using a molecular imprinting method with silica-alumina gel to form complementary structures for template recognition. The molecularly imprinted polymer (MIP) was synthesized by the condensation of silicate from tetraethyl orthosilictate (TEOS) under hydrothermal conditions at 60 degrees C. Aluminum chloride was added as a functional monomer to increase the material's rebinding ability. The selectivity of the MIP receptor prepared with different ratios of template to Si and Al, was examined with seven analytes including: dopamine, epinephrine, norepinephrine, ascorbic acid, homovanillic acid, uric acid, and l-tyrosine. The results showed a size selective effect for the receptors with respect to the recognition of the catecholamines. Some factors affecting the recognition ability were investigated including: the solution pH of analytes, surface capping on the MIP, and the imprinting pH of the silica-alumina solution. Also, the catecholamine MIP films on quartz crystal microbalance (QCM) electrodes were fabricated as sensors for in situ monitoring of the analytes in a 2-propanol solution. PMID- 16257660 TI - Assessment of porous silicon substrate for well-characterised sensitive DNA chip implement. AB - A biochip approach based on porous silicon as substrate is presented. The goal is to enhance the sensitivity of the biochip by increasing the specific surface area on the support. The elaboration of porous silicon layers has been optimized to guarantee good accessibility for large bio-molecule targets. Oligonucleotide probes are synthesised directly on the surface using phosphoramidite chemistry. The high specific surface area of porous silicon allows the direct characterisation, by infrared spectroscopy, of the porous layer formation and the functionalisation steps. The monolayer grafting and derivatisation protocol is additionally characterized by wettability and fluorescence microscopy. The surface modification of porous layers (i.e. thermal oxidation and chemical derivatisation) ensures the stability of the structure against strong chemical reagents used during the direct oligonucleotide synthesis. Finally the protocol is successfully transferred to a flat Si/SiO(2) substrate, and validated by biological target specific recognition during hybridisation tests. In particular, radioactive measurements show a 10-fold enhancement of the oligonucleotide surface density on the porous silicon substrate compared to the flat thermal silica. PMID- 16257661 TI - An electrochemical sensor array system for the direct, simultaneous in vitro monitoring of nitric oxide and superoxide production by cultured cells. AB - A new approach for an amperometric array sensor platform employing arrays of sensors in a 24-well cell culture plate format has been developed for simultaneous in vitro determination of nitric oxide (NO) and superoxide free radicals (O(2)(-)) produced by stimulated cells. The work reported focuses on the direct, real-time monitoring of extracellular production of these two analytes, as well as the effects of their interaction. The sensor platform was manufactured by a combination of sputtering gold electrodes and screen-printing carbon electrodes. The O(2)(-) sensor uses covalent immobilization of cytochrome c via a binder, DTSSP (3,3'-dithio-bis(sulphosuccinimidylpropionate) onto the surface of the Au electrodes, whereas the NO sensor system involves an NiTSPc (nickel tetrasulfonated phthalocyanine) film electrodeposited onto the surface of the carbon electrodes and subsequently covered with an external layer of Nafion. For in vitro demonstration of the platforms as a potential drug-screening system, A172 glioblastoma cells were cultured and transferred into the 24-well arrays. Simultaneous and direct monitoring of NO and O(2)(-) production as a response to chemicals of biomedical relevance was carried out. The results obtained demonstrated that it would be possible to envisage a drug screening platform for compounds designed to be inhibitors of nitric oxide synthase or to have an inhibitory effect on superoxide free radical production. By suitable modification of the electrodes employed it would also be possible to extend the platform to measure alternative species. PMID- 16257662 TI - Direct mediatorless electron transport between the monolayer of photosystem II and poly(mercapto-p-benzoquinone) modified gold electrode--new design of biosensor for herbicide detection. AB - Photosystem II (PSII) modified gold electrodes have been prepared providing mediatorless electron transport on the basis of electrodeposited conductive layer poly-mercapto-p-benzoquinone (polySBQ). Such electrodes are suitable in construction of biosensors for PSII inhibiting herbicides. PolySBQ layer was synthesized on (i) screen-printed gold electrodes and (ii) gold microelctodes in an array on silicon substrate, by electrochemical-oxidation of sulpho-p benzoquinone (SBQ) at +650 mV versus Ag/AgCl. The basic properties of polySBQ layer were characterized using linear sweep voltammetry and atomic force microscopy (AFM). The typical redox response for quinones was observed. The optimal length of the polymer providing direct electron transfer (DET) was found to be very close to 30 nm. PSII particles isolated from the thermophilic cyanobacteria Synechococcus bigranulatus were physically adsorbed on the polySBQ covered gold electrodes. The generation of photocurrent was observed at E=+250 mV (versus Ag/AgCl) without addition of any mediator. The basic properties of DET were studied. We concluded that: (i) PSII active in DET is immobilized in form of monolayer; (ii) the charge transport from PSII to gold working electrode (AuWE) is fast and dominated by the rate of the enzymatic reaction; (iii) polySBQ layer drains electrons from the Q(A) pocket of the photosystem since the electrode activity is inhibited by specific inhibitor, i.e. diuron (DCMU); (iv) the stability of the photosystem immobilized on gold electrodes by using polySBQ is comparable to the stability of PSII in solution under the same experimental conditions; (v) the inhibition of the photosystem by herbicide DCMU follows the sigmoid dependence; (vi) I(50) as well as limit of detection (LOD) show an improved sensitivity compared to other published biosensing systems using PSII as bioactive part. PMID- 16257663 TI - Sub-micron particle behaviour and capture at an immuno-sensor surface in an ultrasonic standing wave. AB - The capture of 200 nm biotinylated latex beads from suspensions of concentration 10(7) to 2.5 x 10(8) particle/ml on an immuno-coated surface of the acoustic reflector in an ultrasound standing wave (USW) resonator has been studied while the acoustic pathlength was less than one half wavelength (lambda/2). The particles were delivered to the reflector's surface by acoustically induced flow. The capture dependencies on suspension concentration, duration of experiments and acoustic pressure have been established at 1.09, 1.46 and 1.75 MHz. Five-fold capture increase has been obtained at 1.75 MHz in comparison to the control (no ultrasound) situation. The contrasting behaviours of 1, 0.5 and 0.2 mum fluorescent latex beads in a lambda/4 USW resonator at 1.46 MHz have been characterized. The particle movements were observed with an epi-fluorescent microscope and the velocities of the particles were measured by particle image velocimetry (PIV). The experiments showed that whereas the trajectories of 1 mum particles were mainly affected by the direct radiation force, 0.5 mum particles were influenced both by the radiation force and acoustic streaming. The 0.2 mum latex beads followed acoustic streaming in the chamber and were not detectably affected by the radiation force. The streaming-associated behaviour of the 200 nm particles has implications for enhanced immunocapture of viruses and macromolecules (both of which are also too small to experience significant acoustic radiation force). PMID- 16257664 TI - Mediatorless voltammetric oxidation of NADH and sensing of ethanol. AB - A simple, selective and sensitive method for the detection of NADH and ethanol is presented. Self-assembled monolayers (SAMs) of mercaptopyrimidine (MPM) and their derivatives, thiocytosine (TC) and 4,6-diamino-2-mercaptopyrimidine (DMP) on gold (Au) electrode are used for the voltammetric detection of NADH and ethanol in neutral aqueous solution. A decrease of 200-300 mV in the overpotential associated with an observable increase in the peak current was obtained for the oxidation of NADH on MPM and TC monolayer-modified electrodes without any redox mediator. The facilitated electron transfer for the oxidation of NADH at the TC monolayer is ascribed to the existence of stable cationic p-quinonoid form of TC. The electrode modified with DMP monolayer could not exhibit stable response for NADH owing to the fouling of electrode surface. The MPM and TC monolayer-modified electrodes show high selectivity and excellent sensitivity (MPM: 0.633+/-0.005 microA cm(-2) microM(-1); TC: 0.658+/-0.008 microA cm(-2) microM(-1)) towards NADH with detection limit (3sigma) of 2.5 and 0.5 microM, respectively. Presence of large excess of ascorbate (AA) does not interfere the detection of NADH and the monolayer-modified electrode shows individual voltammetric peaks for AA and NADH. Voltammetric sensing of ethanol using alcohol dehydrogenase on MPM and TC monolayer-modified electrode is successfully demonstrated and these electrode can detect as low as 0.5 mM ethanol in neutral pH. The sensitivity of the MPM and TC monolayer-modified electrodes toward ethanol was found to be 3.24+/-0.03 and 3.435+/-0.04 microA cm(-2) mM(-1), respectively. PMID- 16257665 TI - Electron transfer mediator micro-biosensor fabrication by organic plasma process. AB - We propose a new strategy for constructing a mediator-type biosensor as a Bio MicroElectroMechanical Systems (BioMEMS) application. A vinylferrocene plasma polymerized film (PPF) was deposited directly onto the surface of an electrode under dry conditions. The resulting redox film was extremely thin, adhered well onto a substrate (electrode), and had a highly crosslinked network structure. This technique, capable of polymeric deposition of any kind of monomer, can also serve the purpose of anti-fouling coating, or layer-to-layer interface creation. With a subsequent plasma process, additional polymeric layer of hydrophilic acetonitrile was superimposed onto the existing vinylferrocene-PPF surface to offer crucial features such that the wettability could be adjusted for a better electron transfer, and amino functional groups could be attached to immobilize a large amount of enzyme. Based upon this scheme, the device fabrication could be designed in a manner that the whole procedure was made up of dry wafer-handling processes, which is compatible with mass production. A prototype device was fabricated to have an array of needle-shaped amperometric micro-biosensors. The resultant thin polymer layer carried a large number of the mediator molecules, accomplishing a lower overpotential (+410 mV) and a rapid response time (<5s). Stressing the advantages of the plasma polymerization process together with some additional features accomplished in our device fabrication, we would discuss new possibilities in the field of BioMEMS. PMID- 16257666 TI - Nanowiring of a redox enzyme by metallized peptides. AB - A molecular assembly consisting of a redox enzyme, NADH peroxidase, a metallized double-helical peptide, and a gold nanoparticle immobilized onto a gold wire derivatized with a benzenedithiol compound, initiated and conducted redox signals in the presence of H(2)O(2) and NADH. The current generated by the binding of NADH, the electron donor, was transduced through the molecular assembly with apparently little loss of signal to the solution. The currents measured correlate to an electron transfer rate constant on the order of 3,000 s(-1) within each assembly. This electron transfer rate is two orders of magnitude higher than the endogenous electron transfer rate from NADH to the native enzyme, 27 s(-1). This rate indicates that the metallized peptide is in a conformation conducive for electron transfer and, in conjunction with the redox enzyme, can form effective conduits of electrical signals. This work demonstrates the feasibility of utilizing designed and highly efficient biomolecular assemblies for the production of ultra-sensitive, in-situ biosensors. PMID- 16257667 TI - Improved strategy for biosensor-based monitoring of water bodies with diverse organic carbon levels. AB - To protect water resources and to control the water quality it is necessary to develop fast, sensitive, cost-effective, and easy-to-use analytical systems, which are able to measure a variety of contaminants in water. Monitoring water bodies with various matrices can be very difficult. The diverse organic carbon level in water samples (e.g. river water or seawater) causes problems at common analysis and in particular at immunological methods. Here, we demonstrate a new method to overcome the partly occurring matrix problems at quasi-continuous real world biosensor monitoring. Therefore, we developed an easy matrix referencing method for our fully automated immunoassays that could be adapted to other applications depending on a similar test-format. The method was developed using a synthetic organic carbon standard, and validated using a diluted turf extract. Results for the ultra-sensitive immunoassay for estrone quantification are shown as example. The developed method was verified using immunoassays for testosterone, progesterone, ethinylestradiol, estradiol, and estriol. PMID- 16257668 TI - The direct electron transfer of glucose oxidase and glucose biosensor based on carbon nanotubes/chitosan matrix. AB - The glucose oxidase (GOD) is entrapped in the composite of carbon nanotubes/chitosan and direct electron transfer reaction between GOD and electrode takes place. The electron transfer rate of GOD is greatly enhanced to 7.73 s(-1) in the system, which is more than one-fold higher than that of flavin adenine dinucleotide adsorbed on the carbon nanotubes (3.1 s(-1)). This maybe results from the conformational change of GOD in the microenvironment enabling the accessibility of active site for GOD to the electrode. Additionally, the bioactivity of GOD modified in the composite on electrode surface is kept. So as prepared electrode can be used as a glucose biosensor exhibiting higher sensitivity (0.5 microA mM(-1)) and better stability. The facile procedure of immobilizing GOD will promote the developments of electrochemical research for protein, biosensors and other bioelectrochemical devices. PMID- 16257670 TI - The evolution of foot morphology in children between 6 and 17 years of age: a cross-sectional study based on footprints in a Mediterranean population. AB - Footprint evaluation is a widely used method for the determination of foot morphology, but its efficacy and validity are considered controversial. Dynamic footprints were obtained from both feet of 5,866 school-aged children (6-17 years old) to detect any foot changes during growth. The interpretation of the imprint was performed using a classification scheme consisting of 6 types of footprints. In this scheme, footprint types I and II represent the typical and intermediate high-arched foot, respectively. Types III and IV represent normal foot variants, while type V corresponds to the low-arched foot and type VI to the severe flat foot, the latter often encountered in pathological conditions. There was statistically significant difference (P<.05) in footprint-type frequencies between boys and girls of ages 7, 9, 11, 14, and 15, which probably indicates the difference in growth potential of the foot between sexes. The proportion of high- and low-arched foot types decreased with increasing age in both boys and girls. Even though critical changes of the foot are believed to occur during pre-school development, this study shows that considerable changes also take place during school age and until late adolescence. PMID- 16257671 TI - Rail external fixation for stabilization of closing base wedge osteotomies and lapidus procedures: a retrospective analysis of sixteen cases. AB - This is a retrospective study of 16 cases of proximal first-ray procedures with rail external fixation. Arthrodesis of the first metatarsal-cuneiform joint was performed on 11 feet, and a closing base-wedge osteotomy was performed on 5 feet. Heel weightbearing in a surgical shoe with ankle joint mobilization was the standard postoperative protocol. Thirteen women and one man with an average age of 41 years (range, 16-64 years) were evaluated, with an average follow-up of 15.12 months (range, 10-24 months). The mean duration of frame application among all patients was 6.75 weeks (range, 2-10 weeks; 7 weeks for the base wedge procedures; 6.64 weeks for the lapidus). Multiple complications developed. Ten patients developed pin-tract infections: 5 required isolated pin removal, whereas 2 required removal of the entire frame because of the severity of infection. Seven patients developed pin loosening. Two patients developed cellulitis requiring long-term intravenous antibiotic therapy. Two nonunions occurred, both with the lapidus procedure, and one was symptomatic. Ten of the 16 patients reported they would have surgery again. Monolateral external rail fixation in combination with heel weightbearing and ankle mobilization demonstrated a high complication rate when used for proximal first-ray procedures. The technique as described is not recommended for routine use with the lapidus and base wedge osteotomy procedures. PMID- 16257672 TI - The Webb-Bannister percutaneous technique for acute Achilles' tendon ruptures: a functional and MRI assessment. AB - This was a study of 57 patients treated for Achilles' tendon rupture between 1994 and 2002: 35 with an open repair and 22 with the Webb-Bannister percutaneous technique. The aim of the study was to evaluate this percutaneous method as compared to other percutaneous and open techniques, with respect to functional result and complications. In addition, postoperative MRI was performed on 40 patients in order to determine whether there were any correlations between clinical results and MRI findings. No significant differences were found between the 2 surgical methods with respect to clinical and functional results, and no wound dehiscence or infections were found in the Webb-Bannister group. There was an 8.6% incidence of wound complications in the open repair group. Similarly, return to work times were not significantly different between the groups (4 months for open repair and 3.75 months for percutaneous repair). Two patients experienced re-rupture after open repair and there was 1 re-rupture following the Webb-Bannister technique. Early weight bearing and the use of the percutaneous repair did correlate to increased postoperative tendon lengthening, though this did not appear to have any clinical consequence. MRI did not appear to be of any value in the clinical or functional evaluation, but it demonstrated a relationship between increased tendon diameter and tendon elongation (P=.0038). In those patients with thicker tendon repair sites, the dorsiflexion tended to exceed the uninjured leg. The functional results of the Webb-Bannister technique were comparable to the open repair. This technique is an effective treatment for acute ruptures less wound complications. PMID- 16257673 TI - Bioabsorbable screws for reduction of Lisfranc's diastasis in athletes. AB - This was a study of 15 athletes who sustained Lisfranc's diastasis and were treated with surgical reduction and fixation with 3.5mm bioabsorbable poly-L lactic acid screws. Patients were prospectively evaluated for their ability to return to activity, maintenance of reduction, and need for implant removal. The average patient age was 28 years; there were 9 females and 6 males. Average followup was 36 months (range: 12-62). The mean return to activity time was 4.2 months (range: 3-6), with a significant difference between males and females (P=.02), 3.4 versus 4.7 months, respectively. The mean preoperative American Orthopedic Foot and Ankle Society midfoot score was 35.4+/-25.0, which increased postoperatively to 92.7+/-7.7 (P=.00001). Overall there were 10 "excellent," 4 "good," and 1 "poor" result. Patients were statistically more likely to have less than an excellent result when diagnosis was delayed by more than 6 weeks (P=0.01). One patient demonstrated a radiographic loss of reduction but returned to full sports activities. The patient with the longest delay in diagnosis demonstrated anatomic reduction but exhibited radiographic degenerative changes and scored a poor result. Three patients required a second surgery for screw or screw head removal. Bioabsorbable screws appear sufficient for treating Lisfranc's diastasis but do not necessarily obviate fixation removal. PMID- 16257674 TI - Implantable electrical bone stimulation for arthrodeses of the foot and ankle in high-risk patients: a multicenter study. AB - This study assessed arthrodesis procedures performed in the foot and ankle of high-risk patients following implantation of an internal electrical bone stimulator. Criteria defining patients as "high risk" included diabetes, obesity, habitual tobacco and/or alcohol use, immunosuppressive therapy, and previous history of nonunion. Standard arthrodesis protocol of bone graft and internal fixation was supplemented with the implantable electrical bone stimulator. A retrospective, multicenter review was conducted of 26 patients (28 cases) who underwent 28 forefoot and hindfoot arthrodeses from 1998 to 2002. Complete fusion was defined as bony trabeculation across the joint, lack of motion across the joint, maintenance of hardware/fixation, and absence of radiographic signs of nonunion or pseudoarthrosis. Radiographic consolidation was achieved in 24 of the 28 cases at an average 10.3+/-4.0 weeks. Followup averaged 27.2 months. Complications included 2 patients who sustained breakage of the cables to the bone stimulator. Five patients underwent additional surgery. Four of the 5 patients had additional surgery in order to achieve arthrodesis. All 4 went on to subsequent arthrodesis. This study demonstrates how arthrodesis of the foot and ankle may be enhanced by the use of implantable electrical bone stimulation. PMID- 16257675 TI - The principles of interference screw fixation: application to foot and ankle surgery. AB - Based on the success of the anterior cruciate ligament model, interference screw fixation is now being applied to a wide variety of orthopedic conditions that require the fixation of tendon or ligament to bone. The primary focus of this article is to present the principles of interference screw fixation. By understanding the principles of interference screw fixation, the foot and ankle surgeon will be able to apply this fixation technique to a wide variety of surgical applications for tendon transfers or ligament repairs. The surgical technique, history, principles of fixation, studies of fixation strength, tissue healing, and foot and ankle indications are reviewed. A modified Girdlestone digital flexor tendon transfer procedure description is included to illustrate how interference screw techniques may be applied to foot surgery. PMID- 16257676 TI - Endometrial carcinoma metastasis to the distal phalanx of the hallux: a case report. AB - Endometrial carcinoma is the most common invasive cancer of the female genital tract and accounts for 7% of all invasive cancer in women. Bony metastasis is uncommon with endometrial carcinoma and distal metastasis is very rare. The purpose of this paper is to present a case of an 86 year-old female with endometrial carcinoma metastasis to the distal phalanx of the hallux. The patient had a known history of endometrial carcinoma with metastases (FIGO IIIC), and had been diagnosed with pulmonary and bony metastases 2 months prior to presentation. Her initial foot complaint was of a painful, infected ingrown toenail. The infection continued to progress following avulsion of the nail, and the patient was then diagnosed with osteomyelitis. Given her past history, the possibility of metastasis to the hallux was also considered. A hallux amputation was performed, and the pathology report revealed the diagnosis of endometrial carcinoma metastasis to the distal phalanx of the hallux. While the amputation site healed uneventfully, the patient refused further treatment measures for her carcinoma and eventually succumbed to the disease. PMID- 16257677 TI - Sterile abscess formation following a corticosteroid injection for the treatment of plantar fasciitis. AB - A sterile abscess following a corticosteroid injection for the treatment of plantar fasciitis has not been reported in the literature. This case report demonstrates such a complication. A 45-year-old man presented with pain, erythemia, edema, and increased warmth to his left foot after 2 cortisone injections into his left heel over a 2-month period for plantar fasciitis. The symptoms began shortly after the second injection and after a 10-day course of oral antibiotics, the condition slowly worsened. A computerized axial tomography scan was suggestive of an abscess. Following incision and drainage, cultures that were taken during the surgical procedure did not produce any organism, prompting a diagnosis of a sterile abscess. The patient was discharged home and recovered from the abscess without reoccurrence or further surgical intervention. PMID- 16257678 TI - Osteoid osteoma of the calcaneus: percutaneous radiofrequency ablation. AB - Osteoid osteoma of the foot can pose particular problems in diagnosis, especially when positioned in a juxta-articular location. It can cause reactive synovitis and simulate arthritis without periostitis. An atypical presentation may delay diagnosis and thus delay treatment. Different modes of treatment have been described including medical management with nonsteroidal, antiinflammatory drugs, and open surgical resection with intralesional, marginal, or wide surgical margins. In recent years, several computed tomography-guided percutaneous techniques have been used to achieve ablation of the nidus with minimal tissue invasion. We report a case of a 39-year-old man with an 8-month history of persistent foot pain who underwent percutaneous radiofrequency ablation of an osteoid osteoma involving the calcaneus. The patient related an immediate relief of pain and had no recurrence of symptoms or the lesion at 3-year follow-up. PMID- 16257679 TI - Foot polydactyly and polysyndactyly: genetic implications in two families. AB - The purpose of this study was to determine the genetic characteristics of foot polydactyly and identify its inheritance pattern by analyzing familial pedigree. Five cases from 2 Korean families were studied: 1 is a family whose members have been affected for 4 generations and the other for 2 generations. Using peripheral blood samples, we performed chromosomal analysis using the banding technique with Giemsa stain and karyotyping. We investigated the shape and structure of 46 chromosomes, looking for translation, deletion, inversion, ring chromosome, and isochromosome abnormalities. All peripheral blood samples demonstrated no chromosomal abnormalities, though the genetic nature of foot polydactyly and a new genetic locus was identified recently by other studies. Familial pedigree analysis suggested that polydactyly was inherited as an autosomal dominant trait in the first family. The mode of inheritance for the second family could not be determined due to an insufficient number of family members. The result of this study brought us to the conclusion that, while genetic factors play a major role in polydactyly, other factors may contribute to its occurrence. PMID- 16257680 TI - Complete medial dislocation of the first cuneiform: a case report. AB - Isolated dislocation of the first cuneiform bone is an extremely rare lesion that can usually be diagnosed on plain radiography. Few cases of this injury have been previously described in the literature. The case presented here resulted from a torsion injury and was treated by means of closed reduction with the aid of pointed reduction forceps. After fixation with Kirschner wires, the foot was immobilized for 6 weeks. The patient recovered uneventfully and remains asymptomatic 3 years later. PMID- 16257681 TI - Occult carcinoma of the lung presenting as pain in the hallux: a case report. AB - Metastatic tumors to the hands and feet (acrometastases) are rare. We report a case in which the primary presentation of a lung carcinoma was a metastatic lesion to the distal phalanx of the hallux. A 60-year-old woman was evaluated for pain in her big toe. Radiographs and computed tomography suggested a benign lesion in the distal phalanx of the hallux, but curettage and biopsy revealed metastatic adenocarcinoma. A chest x-ray revealed a mass in the right perihilar region, which was confirmed by CT, bronchoscopy, and biopsy as carcinoma of the lung. A review of the literature reveals that there is a tendency toward delayed diagnosis in similar cases, especially when the primary lesion is asymptomatic. A high index of suspicion is needed for early diagnosis in such cases. PMID- 16257682 TI - Peroneal tendon repair with autologus hamstring tendons. PMID- 16257683 TI - Interpositional free tendon graft for lesser metatarsophalangeal joint arthropathy. PMID- 16257684 TI - Comment on: "the value of radiographic parameters in the surgical treatment of hallux rigidus" by Zgonis et al, May/June 2005. PMID- 16257686 TI - Optical absorption of tetraphenylporphyrin thin films in UV-vis-NIR region. AB - The optical absorption of thermally evaporated tetraphenylporphyrin (TPP) in the UV-vis-NIR region have been studied. The absorption spectra recorded in the UV vis region for the as deposited and annealed films showed different absorption bands, namely the Soret(B) at region 360-490nm, Q-band region consist of four bands in the region 500-720nm and two other bands labeled N and M in UV region. The Soret band always shows its characteristic effect splitting in all the TPP thin films and the effect of annealing on the intensities of these components have been observed. The spectra of the infrared absorption allow characterization of vibrational modes for the powder, as deposited and annealed thin films. Some of the optical absorption parameters, namely molar extinction coefficient, epsilon, half band width, Deltalambda, electronic dipole strength, q(2) and oscillator strength, f, of the principle optical transitions have also been evaluated. PMID- 16257687 TI - Resolving overlapped spectra with curve fitting. AB - A novel method of curve fitting based on Gaussian function, which is used to resolve the overlapped peaks, is presented in this paper. The resolution of several kinds of overlapped peaks with noise simulated by computer has been performed and discussed in details. This method has been used for resolving of the UV-vis overlapped spectrum. The results indicate that the proposed algorithm can been used to resolve overlapped spectra effectively and satisfactorily. PMID- 16257688 TI - Research on processing medicinal herbs with multi-steps infrared macro fingerprint method. AB - How to apply rapid and effective method to research medicinal herbs, the representative of complicated mixture system, is the current study focus for analysts. The functions of non-processed and processed medicinal herbs are greatly different, so controlling the processing procedure is highly important for guarantee of the curative effect. Almost, the conventional criteria of processing are based on personal sensory experience. There is no scientific and impersonal benchmark. In this article, we take Rehmannia for example, conducting a systematic study on the process of braising Rehmannia with yellow wine by using the multi-steps infrared (IR) macro-fingerprint method. The method combines three steps: conventional Fourier transform infrared spectroscopy (FT-IR), second derivative spectroscopy, and two-dimensional infrared (2D-IR) correlation spectroscopy. Based on the changes in different types of IR spectra during the process, we can infer the optimal end-point of processing Rehmannia and the main transformations during the process. The result provides a scientific explanation to the traditional sensory experience based recipe: the end-point product is "dark as night and sweet as malt sugar". In conclusion, the multi-steps IR macro fingerprint method, which is rapid and reasonable, can play an important role in controlling the processing of medicinal herbs. PMID- 16257689 TI - 1,3-Diaxial steric effects and intramolecular hydrogen bonding in the conformational equilibria of new cis-1,3-disubstituted cyclohexanes using low temperature NMR spectra and theoretical calculations. AB - The conformational equilibria of 3-X-cyclohexanol [X=F (1), Cl (2), Br (3), I (4), Me (5), NMe(2) (6) and MeO (7)] and of 3-X-methoxycyclohexane [X=F (8), Cl (9), Br (10), I (11), Me (12), NMe(2) (13) and MeO (14)] cis isomers were determined from low temperature NMR spectra and PCMODEL calculated coupling constants. The energy differences between aa and ee conformers were obtained from these data (DeltaG(J)(av) and DeltaG(PC)(av), respectively) and also by the additivity principle from data for the monosubstituted cyclohexanes (DeltaG(Ad)). H-1 and H-3 hydrogen vicinal coupling constants and DeltaG(J)(av) values showed that the diequatorial conformer is predominant in the conformational equilibrium of the compounds studied at low temperature. However, DeltaG(PC)(av) data show that compounds 6 and 7 constitute an exception, since they are almost equally populated by ee and aa at room temperature, due to stabilization of their aa conformer by an intramolecular hydrogen bond. DeltaG(Ad) values, obtained according to the additivity principle, show a better agreement for compounds 2 and 3, since the 1,3-diaxial steric effect is counterbalanced by the formation of an intramolecular hydrogen bond (IAHB). For the remaining compounds, DeltaG(Ad) values underestimate the energy differences, since the 1,3-diaxial steric effect, between X and OH or OCH(3), is absent in the monosubstituted compounds used as references. Moreover, the DeltaG(PC)(av), calculated from the coupling constants, obtained through the PCMODEL program, are rather smaller than the DeltaG(J)(av) values, since the program does not have parameters for the effect, observed in this report, of a substituent at gamma position on coupling constants values for the hydrogen under consideration. PMID- 16257690 TI - Near-infrared spectrophotometric determination of compositions of fullerene samples. AB - A novel method has been developed for the sensitive and accurate determination of compositions of fullerene samples. The method is based on the synergistic use of spectrophotometric measurements and partial least square method. The method is not only simple, inexpensive and fast but also is non-destructive. Compositions of various fullerene samples including fullerite which is the precursor to C(60) and C(70), can be directly and non-destructively determined by this method without any time-consuming separation step as in the HPLC method or destruction as in the MS method. PMID- 16257691 TI - NIR spectroscopy of selected iron(II) and iron(III) sulphates. AB - A problem exists when closely related minerals are found in paragenetic relationships. The identification of such minerals cannot be undertaken by normal techniques such as X-ray diffraction. Vibrational spectroscopic techniques may be applicable especially when microtechniques or fibre-optic techniques are used. NIR spectroscopy is one technique, which can be used for the identification of these paragenetically related minerals and has been applied to the study of selected iron(II) and iron(III) sulphates. The near-IR spectral regions may be conveniently divided into four regions: (a) the high wavenumber region>7500 cm( 1), (b) the high wavenumber region between 6400 and 7400 cm(-1) attributed to the first overtone of the fundamental hydroxyl stretching mode, (c) the 5500-6300 cm( 1) region attributed to water combination modes of the hydroxyl fundamentals of water, and (d) the 4000-5500 cm(-1) region attributed to the combination of the stretching and deformation modes of the iron(II) and iron(III) sulphates. The minerals containing iron(II) show a strong, broad band with splitting, around 11,000-8000 cm(-1) attributed to (5)T(2g)-->(5)E(g) transition. This shows the ferrous ion has distorted octahedral coordination in some of these sulphate minerals. For each of these regions, the minerals show distinctive spectra, which enable their identification and characterisation. NIR spectroscopy is a less used technique, which has great application for the study of minerals, particularly minerals that have hydrogen in the structure either as hydroxyl units or as water bonded to the cation as is the case for iron(II) and iron(III) sulphates. The study of minerals on planets is topical and NIR spectroscopy provides a rapid technique for the distinction and identification of iron(II) and iron(III) sulphates minerals. PMID- 16257692 TI - Fe(3+) ions in alkali lead tetraborate glasses--an electron paramagnetic resonance and optical study. AB - Glass systems of composition 90R(2)B(4)O(7)+9PbO+1Fe(2)O(3) (R=Li, Na and K) and 90Li(2)B(4)O(7)+(10-x)PbO+xFe(2)O(3) (x=0.5, 1, 3, 4, 5, 7 and 9 mol %) have been investigated by means of electron paramagnetic resonance (EPR) and optical absorption techniques. The EPR spectra exhibit three resonance signals at g approximately 6.0, 4.2 and 2.0. The resonances at g approximately 6.0 and 4.2 are attributed to Fe(3+) ions in rhombic and axial symmetry sites, respectively. The g approximately 2.0 resonance signal is due to two or more Fe(3+) ions coupled together with dipolar interaction. The EPR spectra of 1 mol % of Fe(2)O(3) doped in lithium lead tetraborate glass samples have been studied at different temperatures (123-433 K). The intensity of g approximately 4.2 resonance signal decreases and the intensity of g approximately 2.0 resonance signal increases with the increase of temperature. The line widths are found to be independent of temperature. The EPR spectra exhibit a marked concentration dependence on iron content. A decrease in intensity for the resonance signal at g approximately 4.2 with increase in iron content for more than 4 mol % has been observed in lithium lead tetraborate glass samples and this has been attributed to the formation of Fe(3+) ion clusters in the glass samples. The paramagnetic susceptibility (chi) is calculated from the EPR data at various temperatures and the Curie constant (C) has been evaluated from 1/chi versus T graph. The optical absorption spectrum of Fe(3+) ions in lithium lead tetraborate glasses exhibits three bands characteristic of Fe(3+) ions in an octahedral symmetry. The crystal field parameter D(q) and the Racah interelectronic repulsion parameters B and C have also been evaluated. The value of interelectronic repulsion parameter B (825 cm( 1)) obtained in the present work suggests that the bonding is moderately covalent. PMID- 16257693 TI - Solid state linear dichroic infrared spectral analysis of benzimidazoles and their N(1)-protonated salts. AB - A stereo structural characterization of 2,5,6-thrimethylbenzimidazole (MBIZ) and 2-amino-benzimidaziole (2-NH(2)-BI) and their N(1) protonation salts was carried out using a polarized solid state linear dichroic infrared spectral (IR-LD) analysis in nematic liquid crystal suspension. All experimental predicted structures were compared with the theoretical ones, obtained by ab initio calculations. The C(s) to C(2v)(*) symmetry transformation as a result of protonation processes, with a view of its reflection on the infrared spectral characteristics was described. PMID- 16257694 TI - A comparative study of molecular complexation of [60]- and [70]fullerenes with 1,3,5-tribromobenzene by UV-vis spectrophotometric method. AB - By UV-vis spectrophotometric method it has been shown that 1,3,5-tribromobenzene (TBB) forms molecular complexes of 1:2 stoichiometry with [60]- and [70]fullerenes. An isosbestic point could be detected in case of the [70]fullerene complex. The formation constant of the [60]fullerene complex is higher than that of the [70]fullerene complex at each of the four temperatures under study. This is in opposite order of the electron affinities of the two fullerenes; moreover, no charge transfer band was observed in the spectra of either complex in solution. This indicates that van der Waals forces, rather than CT interactions, are responsible for complexation. The results reveal that the C atoms at the pentagon vertices of [60]fullerene have greater polarizing power than those in [70]fullerene. PMID- 16257695 TI - Characterisation of prehnite by EPMA, Mossbauer, optical absorption and EPR spectroscopic methods. AB - A sample of prehnite from Rayalaseema zone of Andhra Pradesh, India containing about 2.565 wt.% Fe(2)O(3) is used in the present work. The mineral has been characterized by EPMA, optical absorption, EPR, NIR and Mossbauer techniques. Mossbauer studies confirm the presence of iron as an impurity in two sites. An EPR study on powder sample confirm the presence of Fe(III) impurity in the mineral. Optical absorption spectrum also indicates that Fe(III) impurity is present in two sites with octahedral structure. NIR results are due to water fundamentals. PMID- 16257696 TI - The viscosity and temperature dependence of X-band ESR lineshapes of Gd(III) aqueous complex. AB - X-band ESR spectra of Gd-aqua complex in various weight concentration of glycerol have been recorded at four temperatures. The interpretation of the ESR linewidth is preformed using both the stochastic Liouville approach (SLA) and a perturbation theory. The SLA uses a one dynamic model of the zero-field splitting whereas the perturbation approach uses a two dynamic model. Both models can reproduce the variation of the linewidth with respect to viscosity. In the SLA model, both the zero-field splitting (ZFS) interaction and the correlation time vary with the glycerol content. In the two dynamic perturbation model, only the correlation times are viscosity dependent. The two models give different NMRD profiles. PMID- 16257697 TI - DFT study on the geometric, electronic structure and Raman spectra of 5,15 diphenylporphine. AB - The ground state geometric, electronic structure and Raman spectra of 5,15 diphenylporphine (H(2)DPP) have been studied using B3LYP/6-31G(d) method and compared with that of well-studied free base porphine (H(2)P) and meso tetraphenylporphine (H(2)TPP). Calculation shows that 5,15-substitution causes remarkable in-plane distortion, whereas the resulting out-of-plane distortion is negligible. The calculated electronic structure of H(2)DPP is consistent with the absorption spectra compared with H(2)P and H(2)TPP. The calculated vibrational frequencies of H(2)DPP scaled with a single factor of 0.971 agree well with experimental data (the rms error is 8.0 cm(-1)). The assignment of experimental Raman bands of H(2)DPP was discussed on the basis of theoretical calculation and the comparison with that of H(2)P and H(2)TPP. The splitting of some vibrational modes involving the motion of C(m) atom, such as nu(1), nu(8), and nu(10), was observed and was attributed to the diversification of the environment around C(m) atoms. As the shift of absorption peaks, the shift of some structure-sensitive Raman bands of H(2)DPP form that of H(2)TPP and H(2)P was attributed to the in plane nuclear reorganization (IPNR) induced by phenyl-substitution, though the contribution of nonplanarity mechanism could not be excluded completely. PMID- 16257698 TI - The structure and the Raman vibrational spectrum of the beryllium aquacation. AB - The experimental Raman vibrational spectrum of the 5.94 m water solution of the beryllium(II) chloride has been acquired. Theoretical frequencies, infrared and Raman intensities of the vibrational spectrum of the beryllium cation tetrahydrate have been calculated by means of quantum chemical approach. The peaks of the experimental spectrum have been assigned on the basis of the results of the quantum-chemical calculations. It has been shown that the hydrating surrounding of the aquacation increases effectively the frequency of the beryllium-oxygen stretching vibration by 16% in comparison with the free complex. PMID- 16257699 TI - Contrast effect of hydrogen bonding on the acceptor and donor OH groups of intramolecularly hydrogen-bonded OH pairs in diols. AB - We studied the influence of hydrogen bonding on the fundamental and overtone bands of the OH-stretching vibration of each OH group in the intramolecularly hydrogen-bonded OH(I)::OH(II) pair in 1,2-, 1,3- and 1,4-diols. The hydrogen bonding between the two OH groups significantly increases in strength from the five-membered ring of a 1,2-diol to the seven-membered ring of a 1,4-diol. Although the hydrogen bonding does not affect the vibrational property of the OH(II) (or acceptor), it significantly influences the OH(I) (or donor). As the hydrogen bonding becomes stronger from a 1,2- to a 1,4-diol, the fundamental band of the OH-stretching shifts downwards by from about 50 to 140 cm(-1), and the overtone band markedly decreases in intensity, although the effect on the intensity and bandwidth of the fundamental band varies among 1,2-, 1,3- and 1,4 diols. The quantum-mechanically calculated normal frequencies of the acceptor and donor OH groups in the hydrogen-bonded ring are in good agreement with the observed frequencies. The calculated interatomic distance between the O of an acceptor OH and the H of a donor OH is the shortest for a 1,4-diol, which is consistent with the largest frequency shift caused by the hydrogen bonding. PMID- 16257700 TI - Solid state (13)C NMR and FT-IR spectroscopy of the cocoon silk of two common spiders. AB - The structure of the silk from cocoons of two common spiders, Araneus diadematus (family Araneidae) and Achaearanea tepidariorum (family Theridiidae) was investigated by means of (13)C solid state NMR and FT-IR spectroscopies. The combined use of these two techniques allowed us to highlight differences in the two samples. The cocoon silk of Achaearanea tepidariorum is essentially constituted by helical and beta-sheet structures, whereas that of Araneus diadematus shows a more complex structure, containing also beta-strands and beta turns. Moreover, the former silk is essentially crystalline while the latter contains more mobile domains. The structural differences of the two cocoon silks are ascribed to the different habitat of the two species. PMID- 16257701 TI - To what extent can methyl derivatives be regarded as stabilized tautomers of xanthine? AB - Taking xanthine (Xan) as an example, validity of an approach to experimental investigations of nucleotide bases' tautomeric equilibrium, based on the use of methyl derivatives corresponding to their prototropic tautomers, was studied by (1)H NMR in dimethylsulfoxide (DMSO) and by quantum chemical calculations at the B3LYP/6-311++G(d,p) level of theory. From (1)H NMR spectra of m(7)Xan, m(9)Xan, Xan and m(3)Xan conclusion was made that the N7H tautomeric forms of the last two compounds dominate in solution, which was supported by quantum chemical data. Calculated relative energies of the N9H tautomers of Xan and m(3)Xan (8.74 and 9.57 kcal/mol, accordingly) are rather close to the m(9)Xan one (9.11 kcal/mol). Nonspecific influence of DMSO modelled by the COSMO algorithm therewith reduces these values by approximately 2-3 kcal/mol. The data obtained imply that methyl derivatives are rather good models of high-energy tautomers of nucleotide bases, if their relative energies are not less than a few kcal/mol. PMID- 16257703 TI - Proton transfer reaction of a new orthohydroxy Schiff base in some protic and aprotic solvents at room temperature and 77 K. AB - The ground and excited state proton transfer reactions of a new orthohydroxy Schiff base, salicylidine-3,4,7-methyl amine (SMA) has been studied by means of absorption, emission and time resolved fluorescence spectroscopy in some polar protic and aprotic solvents at room temperature and 77K. The spectral behavior of SMA has been investigated both in neutral and basic conditions. The intramolecularly hydrogen bonded enol and zwitterion have been detected in pure methanol and ethanol, the anion is detected in basic condition. At 77K, SMA shows phosphorescence in pure methanol and ethanol. From nanosecond measurements and quantum yields of fluorescence, we have estimated the decay rates of proton transfer reaction in methanol and ethanol. PMID- 16257702 TI - The synthesis and the solvent and substituent effect on the spectroscopic characteristic of 3-ethyl-2-(p-substitued styryl)benzothiazolium iodides. AB - The photophysical and photochemical properties of p-substitued 2-styryl ethylbenzothiazolium iodides, possessing different electron-withdrawing or electron-donating groups are described. The dyes were prepared by the condensation of 3-ethyl-2-methylbenzothiazole salts with p-substituted benzaldehydes. The synthesis of suitable substrates is presented as well. We describe here the absorption, emission spectra and the luminescence quantum yield of hemicyanine dyes (SH) measured in 11 different organic solvents of varying polarity. Molecular structure of the synthesized dyes was established by (1)H NMR, electronic absorption and fluorescence spectrometry. The spectral data confirmed that all the compounds exist in E-configuration of their styryl residues. The planar molecular conformation is typical for the compounds with five-membered side aromatic moieties (for example benzothiazole). The compounds possessing N-alkyl substituent in phenyl ring, in contrast to the compounds with other substituents, exhibit low fluorescence quantum yield in THF solution. This indicates that for N-alkyl derivatives the non-radiative processes are much more effective than the radiative ones. The electronic absorption and fluorescence emission spectra of tested dyes demonstrate high sensitivity to the nature of substituent introduced into the aromatic ring. PMID- 16257704 TI - Infrared and electronic spectra of copper (II) complex of maleonitriledithiolate and 4,4'-dimethyl-2, 2'-bipyridine and their theoretical studies. AB - The molecular structure and binding, as well as infrared and electronic spectroscopic properties for the title complex Cu(mnt)(dmbpy)(mnt(2 )=maleonitriledithiolate, dmbpy=4,4'-dimethyl-2,2'-bipyridine) were studied in this paper. With semi-empirical PM3 and non-empirical DFT (B3LYP/6-311G*) methods, the molecular geometry of the complex was optimized and corresponding vibrational spectra in the gaseous state were obtained. The calculated results derived from DFT were more reasonable than those from PM3. The point group of Cu(mnt)(dmbpy) in isolated gaseous state was C(2), in which Cu(II) adopted a distorted tetrahedral geometry and the dihedral angle between the N(2)Cu and S(2)Cu planes was about 29.814 degrees. And a complete assignment to the IR spectra of such a complicated molecule was exhibited. With ZINDO/S method an electronic spectrum was calculated. The results showed that the calculated values generally agreed with the observed ones. And a detailed explain was made on its electronic spectra. PMID- 16257705 TI - Preparation and spectroscopic studies on charge-transfer complexes of 2,2' bipyridine with picric and chloranilic acids. AB - Charge-transfer (CT) complexes formed on the reaction of 2,2'-bipyridine with some acceptors such as picric acid (HPA) and chloranilic acid (H(2)CA) have been studied in CHCl(3) and MeOH at room temperature. Based on elemental analysis and IR spectra of the solid CT complexes along with the photometric titration curves for the reactions, the data obtained indicate the formation of 1:1 charge transfer complexes [(bpyH)(PA)] and [(bpyH(2))(CA)], respectively. The infrared and (1)H NMR spectroscopic data indicate a charge-transfer interaction associated with a proton migration from the acceptor to the donor followed by intramolecular hydrogen bonding. The formation constants (K(C)) for the complexes were shown to be dependent on the structure of the electron acceptors used. PMID- 16257707 TI - EPR of vanadyl impurity in zinc ammonium trihydrogen bis(orthophosphate) monohydrate single crystal. AB - The electron paramagnetic resonance spectroscopic investigation of VO(II)-doped zinc ammonium trihydrogen bis(orthophosphate) monohydrate single crystal has been carried out at 300K. The single crystal, rotated along the three orthogonally crystallographic axes, have yielded spin Hamiltonian parameters g and A as: g(xx)=1.978, g(yy)=1.974, g(zz)=1.925 and A(xx)=7.4, A(yy)=7.8, A(zz)=19.4 mT. These spin Hamiltonian parameters reflect a slight deviation from axially symmetry to rhombic, which is elucidated by the interstitial occupation of the vanadyl ion. The isofrequency plots and EPR spectra at few orientations have been simulated using the calculated spin Hamiltonian parameters. The Fermi contact parameter (kappa) and dipolar interaction parameter (P) have been calculated as 0.857 and -128 x 10(-4)cm(-1), respectively. The percentage of metaloxygen bond has been identified as 20%. PMID- 16257706 TI - Spectroscopic properties and Judd-Ofelt theory analysis of erbium chelates. AB - Erbium chelates including tris(acetylacetonato) erbium(III) monohydrate, tris(acetylacetonato)(1,10-phenanthroline) erbium(III) and tris(trifluoroacetylacetonato)(1,10-phenanthroline) erbium(III) are synthesized. Judd-Ofelt theory is employed on basis of the UV-Vis-NIR absorption spectra of erbium chelates dissolved in methanol. Judd-Ofelt parameters of erbium chelates are determined by a least square fitting and dealt with the chemical structure of erbium chelates. Photoluminescence characteristics of erbium chelates are investigated upon excitation at 488 nm by an Ar(+) laser. The qualitative correlation of Judd-Ofelt parameters with photoluminescence properties for erbium chelates is also discussed. It is found that larger Omega(6) value for erbium chelate is and larger photoluminescence intensity at 1.54 microm is, and Omega(2) value should contribute to the photoluminescence full width at half maximum (FWHM) at 1.54 microm. The changes of Judd-Ofelt parameters result from the introduction of the second ligand phenathroline or the substitution of electron drawing group CF(3) in beta-diketone for erbium chelates. PMID- 16257708 TI - The phase transition temperatures of a liquid crystal determined from FT-IR spectra explored by principal component analysis. AB - The FT-IR spectra of a thin layer of pure 4-chloro-2'-hydroxy-4' pentyloxyazobenzene (CHPAB) were studied as a function of temperature. A detailed analysis of the intensity variations was performed by a method based on principal component analysis (PCA). It was shown that the phase transition temperatures obtained by means of PCA and those determined by differential scanning calorimetry (DSC), the most widely used technique in the field, were nearly identical. The PCA results revealed that the transition from solid to a liquid crystalline (LC) phase (smectic A) is more drastic phase transition in terms of infrared absorption changes. The nematic to isotropic phase transition is much less infrared sensitive. Very much smaller absorption changes are associated with the transition between the smectic and nematic mesophases. The pattern of the intensity changes strictly is correlated with the orientation of the CHPAB molecules towards the surface windows due to the surface-induced homeotropic alignment of LC molecules. The important role of hydrogen bonding interaction on the observed transition is disclosed. PMID- 16257709 TI - Infrared and laser Raman spectral studies of bis(dl-aspartic acid) sulfate. AB - The infrared and Raman spectra of bis(dl-aspartic acid) sulfate in the crystalline state are recorded at room temperature and the vibrational assignments of the observed wavenumbers are made. The presence of two carbonyl (CO) groups has been identified in the title complex and the two carbonyl groups are in different environment. The extensive intermolecular hydrogen bonding in the crystal has been identified by the shifting of bands due to the stretching and bending modes of the various functional groups. The sulfate group forms the anion. Its fundamentals continue to be degenerate except for the asymmetric bending mode and it suggests that the T(d) symmetry of SO(4)(2-) group has not been affected in the crystal. PMID- 16257710 TI - Raman spectroscopy of the mineral rhodonite. AB - The mineral rhodonite an orthosilicate has been characterised by Raman spectroscopy. The Raman spectra of three rhodonites from Broken Hill, Pachapaqui and Franklin were compared and found to be similar. The spectra are characterised by an intense band at around 1000 cm(-1) assigned to the nu(1) symmetric stretching mode and three bands at 989, 974 and 936 cm(-1) assigned to the nu(3) antisymmetric stretching modes of the SiO(4) units. An intense band at around 667 cm(-1) was assigned to the nu(4) bending mode and showed additional bands exhibiting loss of degeneracy of the SiO(4) units. The low wave number region of rhodonite is complex. A strong band at 421.9 cm(-1) is attributed to the nu(2) bending mode. The spectra of the three rhodonite mineral samples are similar but subtle differences are observed. It is proposed that these differences depend upon the cationic substitution of Mn by Ca and/or Fe(2+) and Mg. PMID- 16257711 TI - Raman spectroscopy of halotrichite from Jaroso, Spain. AB - Raman spectroscopy complimented with infrared ATR spectroscopy has been used to characterise a halotrichite FeSO(4) x Al(2)(SO(4))(3) x 22 H(2)O from The Jaroso Ravine, Aquilas, Spain. Halotrichites form a continuous solid solution series with pickingerite and chemical analysis shows that the jarosite contains 6% Mg(2+). Halotrichite is characterised by four infrared bands at 3569.5, 3485.7, 3371.4 and 3239.0 cm(-1). Using Libowitsky type relationships, hydrogen bond distances of 3.08, 2.876, 2.780 and 2.718 Angstrom were determined. Two intense Raman bands are observed at 987.7 and 984.4 cm(-1) and are assigned to the nu(1) symmetric stretching vibrations of the sulphate bonded to the Fe(2+) and the water units in the structure. Three sulphate bands are observed at 77K at 1000.0, 991.3 and 985.0 cm(-1) suggesting further differentiation of the sulphate units. Raman spectrum of the nu(2) and nu(4) region of halotrichite at 298 K shows two bands at 445.1 and 466.9 cm(-1), and 624.2 and 605.5 cm(-1), respectively, confirming the reduction of symmetry of the sulphate in halotrichite. PMID- 16257712 TI - Raman spectroscopy of newberyite, hannayite and struvite. AB - The phosphate minerals hannayite, newberyite and struvite have been studied by Raman spectroscopy using a thermal stage. Hannayite and newberyite are characterised by an intense band at around 980cm(-1) assigned to the v(1) symmetric stretching vibration of the HPO(4) units. In contrast the symmetric stretching mode is observed at 942cm(-1) for struvite. The Raman spectra are characterised by multiple v(3) anti-symmetric stretching bands and v(2) and v(4) bending modes indicating strong distortion of the HPO(4) and PO(4) units. Hannayite and newberyite are defined by bands at 3382 and 3350cm(-1) attributed to HOPO(3) vibrations and hannayite and struvite by bands at 2990, 2973 and 2874 assigned to NH(4)(+) bands. Raman spectroscopy has proven most useful for the analysis of these 'cave' minerals where complex paragenetic relationships exist between the minerals. PMID- 16257713 TI - Simultaneous determination of cobalt, nickel and palladium in micellar media using partial least square regression and direct orthogonal signal correction. AB - A simple, novel and sensitive spectrophotometric method was described for the simultaneous determination of cobalt, nickel and palladium. The method is based on the complex formation of Co, Ni and Pd with 1-(2-pyridylazo)-2-naphtol (PAN) in Tween-80 micellar media. All factors affecting on the sensitivity were optimized and the linear dynamic range for determination of Co, Ni and Pd was found. The experimental calibration matrix was designed by measuring the absorbance over the range of 520-700 nm for 21 samples of 0.10-1.0, 0.050-0.50 and 0.050-4.0 microg ml(-1) of Co, Ni and Pd, respectively. The partial least square (PLS) modeling based on singular value decomposition (SVD) was used for the multivariate calibration of the spectrophotometric data. The direct orthogonal signal correction was used for pre-processing of data matrices and the prediction results of model, with and without using direct orthogonal signal correction, were statistically compared. The effects of various anions and cations on selectivity of the method were investigated. The proposed method was successfully applied to the determination of Co, Ni and Pd in water and alloy samples. PMID- 16257714 TI - Spectrophotometric study of the inclusion complex between beta-cyclodextrin and dibenzoyl peroxide and its analytical application. AB - The noncovalent interaction of dibenzoyl peroxide and beta-cyclodextin (beta-CD) has been studied by spectrophotometry. The mechanism of the inclusion was studied. The results showed that beta-CD reacts with dibenzoyl peroxide to form a 2:1 host-guest complex with an apparent formation constant of 2.5 x 10(4)mol( 2)L(2). The beta-CD reacts with benzoic acid to form a 1:1 host-guest complex with an apparent formation constant of 6.9 x 10(2)mol(-1)L after the dibenzoyl peroxide was reduced by hydroxyl ammonium. Based on the enhancement of the absorbance of dibenzoyl peroxide produced through complex formation, a spectrophotometric method for determination of dibenzoyl peroxide in bulk aqueous solution in the presence of beta-CD was developed. A linear relationship between the absorbance and dibenzoyl peroxide concentration was obtained in the range of 0.300-50.0 microg mL(-1). Linear regression equation of the calibration graph C=0.02926+53.25 A, with a correlation coefficient of 0.9984 and a relative standard deviation (R.S.D.) of 3.4%. The detection limit was 0.200 microg mL(-1), and the recovery was from 98.00 to 105.0%. The proposed method was used to determine the dibenzoyl peroxide in the flour with satisfactory results. The principal advantage of the proposed method is its excellent selectivity based on molecule recognition of beta-CD. PMID- 16257716 TI - A spectroscopic study on applicability of spectral analysis for simultaneous quantification of l-dopa, benserazide and ascorbic acid in batch and flow systems. AB - The usefulness of derivative spectrophotometry for simultaneous assay of l-dopa, benserazide and ascorbic acid in pharmaceuticals was studied. The parameters of derivatisation depends on composition of solution in which particular compound was determined. For quantification of l-dopa in mixtures with benserazide or ascorbic acid the first derivative was used. Its determination in ternary mixture (l-dopa+benserazide+ascorbic acid) is possible by third derivative spectra. Benserazide was assayed in presence of l-dopa using first derivative while in ternary mixture by third derivative. Direct determination of ascorbic acid is possible applying first derivative only in presence of l-dopa. The elaborated derivative spectrophotometric methods were used for assaying of l-dopa and benserazide in their commercial form "Madopar". The proposed spectrophotometric derivative method of simultaneous determination of l-dopa and benserazide was combined with FIA technique. PMID- 16257715 TI - Spectroscopic characterization of effective components anthraquinones in Chinese medicinal herbs binding with serum albumins. AB - The interactions of serum albumins such as human serum albumin (HSA) and bovine serum albumin (BSA) with emodin, rhein, aloe-emodin and aloin were assessed employing fluorescence quenching and absorption spectroscopic techniques. The results obtained revealed that there are relatively strong binding affinity for the four anthraquinones with HSA and BSA and the binding constants for the interactions of anthraquinones with HSA or BSA at 20 degrees C were obtained. Anthraquinone-albumin interactions were studied at different temperatures and in the presence of some metal ions. And the competition binding of anthraquinones with serum albumins was also discussed. The Stern-Volmer curves suggested that the quenching occurring in the reactions was the static quenching process. The binding distances and transfer efficiencies for each binding reactions were calculated according to the Foster theory of non-radiation energy transfer. Using thermodynamic equations, the main action forces of these reactions were also obtained. The reasons of the different binding affinities for different anthraquinone-albumin reactions were probed from the point of view of molecular structures. PMID- 16257717 TI - A scaled quantum mechanical force field for the sulfuryl halides II. The SO(2)XF (X=Cl, Br) halides. AB - Force fields and vibrational wavenumbers were calculated for the molecules SO(2)XF (X=Cl, Br) using DFT techniques. The previously available experimental data and assignments for SO(2)ClF and SO(2)BrF were partially confirmed by the theoretical results. These data were subsequently used in the definition of scaled quantum mechanics force fields for such molecules. A comparison of the obtained force constants is made with results previously published for the SO(2)X(2) molecules. PMID- 16257718 TI - Iron pentacarbonyl detection limits in the cigarette smoke matrix using FT-IR spectroscopy. AB - Endogenous metals present in tobacco from agricultural practices have been purported to generate metal carbonyls in cigarette smoke. Transition metal catalysts, such as iron oxide, have been investigated for the reduction of carbon monoxide (CO) in cigarette smoke. These studies motivated the development of an analytical method to determine if iron pentacarbonyl [Fe(CO)(5)] is present in mainstream smoke from cigarette models having cigarette paper made with iron oxide. An FT-IR puff-by-puff method was developed and the detection limit was determined using two primary reference spectra from different sources to estimate the amount of Fe(CO)(5) present in a high-pressure steel cylinder of CO. We do not detect Fe(CO)(5) in a single 35 mL puff from reference cigarettes or from those cigarette models having cigarette paper made with iron oxide, with a 30 ppbV limit of detection (LOD). Also, it was shown that a filter containing activated carbon would remove Fe(CO)(5). PMID- 16257719 TI - Spontaneous self-association of adenine and uracil in polycrystals from low temperature FTIR spectra in the range below 1000 cm(-1). AB - FTIR spectra of solid samples of co-crystallized adenine and uracil were measured at 10K in the range below 1000cm(-1). New bands ascribable to the N3H (uracil) and NH(2) (adenine) out of plane vibrations, which disappear upon D-exchange, were revealed in comparison with the spectra of pure polycrystalline adenine and uracil obtained in the same conditions. The observed changes relate to the same groups that establish the H-bonds in base pairs of naturally occurring nucleic acids, despite the presence of an extra proton donor NH-group in both molecules. The well-established empirical correlation between the out of plane NH vibrational frequencies and H-bond energies was successfully applied for estimation of the latter in the mixed crystal. PMID- 16257720 TI - Abnormal FT-IR and FTRaman spectra of ionic liquids confined in nano-porous silica gel. AB - A new concept of designing and synthesizing highly dispersed ionic liquids was developed through physical confinement or encapsulation of them into silica gel matrix with sol-gel process. A series of silica gel confined ionic liquids were synthesized through this process and characterized by diffuse-reflectance infrared Fourier transform spectroscopy (DRIFTS) and FTRaman analysis, and abnormal FT-IR and FTRaman spectra were observed. The silica gel matrixes confined ionic liquid BMImBF(4) were further characterized by BET analysis after the ionic liquid was almost completely washed out by acetone under refluxing conditions and meso-porous silica gel matrixes were obtained according to the N(2) adsorption measurements, which suggested that the particle-size of the dispersed ionic liquids was in nano-scale. In consideration of the results obtained together, it could be found that the abnormal FT-IR and FTRaman spectra were changed with the pore-size of the silica gel matrix. For example, obvious abnormal FT-IR and FTRaman spectra appeared when the particle-size of ionic liquid BMImBF(4) is smaller than 11 nm while they disappeared again if the corresponding particle-size >11 nm. These results indicated that nano-effect, or restriction effect, produced from the nano-pores of silica gel was the reason for the abnormal FT-IR and FTRaman spectra. PMID- 16257721 TI - A spectroscopic study of the inclusion of azulene by beta- and gamma cyclodextrins. AB - The inclusion of azulene (AZ) inside the cavities of beta-cyclodextrin (beta-CD) and gamma-cyclodextrin (gamma-CD) was studied using absorption, fluorescence and induced-circular dichroism spectroscopy. The inclusion of AZ into the cavity of beta-CD has a stoichiometry of 1:1, whereas that of AZ/gamma-CD complex is 1:2. The equilibrium constants for the formation of the two complexes were calculated to be 780+/-150 M(-1) for AZ:beta-CD and (4.5+/-0.86)x10(5) M(-2) for AZ:(gamma CD)(2). The latter is due to a stepwise equilibrium mechanism in which a 1:1 complex is formed with a binding constant of 775 M(-1), followed by the formation of a 1:2 complex with a binding constant of 580 M(-1). The difference between the two binding constant values is slight, indicating an almost equal contribution from each of the gamma-CD molecules to the overall binding in AZ:(gamma-CD)(2). From the induced-circular dichroism spectra, the inclusion of AZ was found to be axial in AZ:beta-CD and nearly axial in AZ:(gamma-CD)(2). PMID- 16257722 TI - Experimental and theoretical study of 2,5-diaryloxazoles whose aryl are para substituted phenyl groups. AB - Absorption and emission properties of some phenoxy derivatives of diphenyloxazole (PPO) are presented and discussed. The photophysical properties reflect a dependence on the substituted 2 or 5 position of the oxazole ring. The experimental data were correlated with molecular parameters such as molecular flexibility, electronic structure and the singlet-triplet gap. The substitution with heteroatomic-bridged phenyls maintains the same frontier orbitals (m, m+1) as in the parent compound, but introduces a new molecular orbital, m-1, located on the terminal phenyl. The presence of the so-called band B in the absorption spectra of some diaryloxazoles was attributed to the m-1-->m+1 transition participating to the second excited state wave function. The other main component of this state, the m-->m+2 transition, was found to have a forbidden character, explaining the lack or the low intensity of band B. The decrease of the fluorescence quantum yield subsequent to substitution of PPO with phenoxy fragments was found to be due to the enhanced molecular flexibility comparing to PPO. The differences between the 2- or 5-substituted derivatives were rationalized in terms of a smaller S(1)-T(2) gap for the former, thus increasing the rate of the overall nonradiative intersystem crossing processes. PMID- 16257724 TI - Spectroscopic study on the photophysical properties of novel lanthanide complexes with long chain mono-L phthalate (L=hexadecyl, octadecyl and eicosyl). AB - Ortho-phthalic anhydride was modified with long chain alcohol (1-hexadecanol, 1 octadecanol and 1-eicosanol) to their corresponding mono-L phthalate (L=hexadecyl, octadecyl and eicosyl), i.e. monohexadecyl phthalate (16-Phth), monooctadecyl phthalate (18-Phth), and monoeicosyl phthalate (20-Phth), respectively. Nine novel lanthanide (Eu(3+), Tb(3+) and Dy(3+)) complexes with these three mono-L phthalate ligands were synthesized and characterized by elemental analysis and IR spectra. The photophysical properties of these complexes were studied in detail with various spectroscopes such as ultraviolet visible absorption spectra, low temperature phosphorescence spectra and fluorescent spectra. The ultraviolet-visible absorption spectra show some band shifts with the different chain-length of phthalate monoester and homologous lanthanide complexes. From the low temperature phosphorescent emission, the triplet state energies for these three ligands were determined to be around 22,650 cm(-1) (16-Phth), 23,095 cm(-1) (18-Phth) and 22,400 cm(-1) (20-Phth), respectively, suggesting they are suitable for the sensitization of the luminescence of Eu(3+), Tb(3+) and Dy(3+). The fluorescence excitation and emission spectra for these lanthanides complexes of the three ligands take agreement with the above predict from energy match. PMID- 16257723 TI - Characterization and biological activities of two copper(II) complexes with diethylenetriamine and 2,2'-bipyridine or 1,10-phenanthroline as ligands. AB - Two new mixed ligand copper(II) complexes with diethylenetriamine, 2,2' bipyridine and 1,10-phenanthroline have been synthesized. The crystal and molecular structures of [Cu(dien)(phen)](ClO(4))(2) and [Cu(dien)(bipy)](BF(4))(2) (dien=diethylenetriamine, phen=1,10-phenanthroline, bipy=2,2'-bipyridine) were determined by X-ray crystallography from single crystal data. These two complexes have similar structures. The EPR spectral data also suggest that these complexes have distorted square pyramidal geometry about copper(II). Anti-microbial and superoxide dismutase activities of these complexes have also been measured. They show the higher SOD activity than the corresponding simple Cu(II)-dien/Cu(II)-PMDT (PMDT=N,N,N',N',N''-pentamethyldiethylenetriamine) complexes because of a strong axial bond of one of the nitrogen atoms of the alpha-diimine. Both the complexes have been found to cleave plasmid DNA in the presence of co-reductants such as ascorbic acid and glutathione. PMID- 16257725 TI - Termination and side-reactions of polystyryllithium with beta-bromostyrene. AB - Low molecular weight polystyryllithium was synthesised and terminated with a two fold molar excess of beta-bromostyrene to produce an end-functionalised polymer. 1,3-Diphenylallyl-terminated polystyrene and other reaction products were analysed by FT-Raman spectroscopy. By comparison with model compounds, it can be deduced that beta-bromostyrene with a predominantly trans configuration gave rise to primarily cis conformational unsaturation at the polystyrene chain-end. It was also demonstrated that a polystyrene 'dimer' and 1,4-diphenylbutadiene were produced via side-reactions which are considered to result from metal-halogen exchange in the reagents. PMID- 16257726 TI - Gas phase infrared spectrum and ab initio calculations of phosphorus(III) thiocyanide, SPCN. AB - An attempt has been made to record the gas phase infrared spectrum of phosphorus(III) thiocyanide, SPCN, for the first time. The molecule was generated by an on-line process using phosphorus(III) thiochloride, SPCl, as a precursor passed over heated silver cyanide at about 350 degrees C. The products were characterized by the infrared spectra of their vapors. The low resolution gas phase Fourier transform infrared spectrum shows three of six characterized fundamental modes of SPCN within the range of the spectrometer used at 2151, 743 and 622 cm(-1) These three bands were assigned to nu(1)(C[triple bond]N stretch), nu(2)(S=P stretch), and nu(3)(C--P stretch), respectively. Ab initio self consistent-field (SCF) molecular orbital (MO) and Moller-Plesset second-order perturbation theory (MP2) calculations were performed to determine the geometry, total energy and the vibrational frequencies of SPCN. PMID- 16257727 TI - Does phosphoryl protonation occurs in aqueous phosphoesters solutions. AB - Ionisation of trimethylphosphate (TMP), dimethylphosphate (DMP) and diethylphosphate (DEP) is investigated by acidic titration in water by Raman (R), Fourier transform infrared (FTIR) and nuclear magnetic resonance (NMR) spectroscopies. The vibrational frequencies of the PO(2)(-) ionic form and the neutral form were found in accord with the literature. While increasing further H(+) concentration, the PO band disappears in the benefit of new ones. These results, together with deuteration experiments indicate the presence of a new ionic form positively charged with general formula R(1)R(2)R(3)P(OH)(+) or R(1)R(2)P(OH)(+)(2). The pK of this phosphonium entities is lying in the range 2, -4. These results were confirmed by (31)P NMR titration. The occurrence of such a phosphonium ion in aqueous solutions might be of crucial importance for biochemical reactions and interactions, owing to the large spread of phosphoryl group in biomolecules and keeping in mind that intracellular compartments are more likely concentrated media with little free water than real aqueous solutions. Furthermore, pK's can be shifted by physical-chemical parameters like dielectric constant and electric field. This may involve at least fractional positive charge apparition that might be important in biochemical regulation by charge-charge and charge-dipole interactions. This finding will gain to be further explored on more complex molecules like phospholipids, nucleic acids and proteins. PMID- 16257728 TI - Vibrational spectrum of chlorotrimethylsilane. AB - The IR and Raman spectra of chlorotrimethylsilane (CTMS) were re-examined. Gas and liquid phase spectra were newly recorded and the analysis of the second derivatives of intensities in some complex spectral regions was carried out. The experimental study was combined with theoretical calculations at the B3LYP level with two different basis sets, i.e., 6-31G(*) and DZP+diff. The use of the SQM methodology, allowed us to review the assignment of the bands for nu(3) (methyl deformation), nu(5) (methyl rocking) and nu(8) (SiC deformation) normal modes of A(1) symmetry, as well as nu(16), nu(17) (methyl deformations) and nu(20) (methyl rocking) normal modes of E symmetry. The description of the asymmetric CH stretchings spectral region has been improved and, in addition, new values of vibrational frequencies were predicted for A(2) normal modes, inactive in both IR and Raman spectra. The equilibrium geometry parameters of CTMS obtained in this work at the MP2/DZP+diff level are closer to the experimental values than previous predictions. PMID- 16257729 TI - Spectroscopic properties of Pr(3+) doped in tellurite glass. AB - The absorption and fluorescence spectra of Pr(3+) doped in tellurite glass has been recorded and analyzed in terms of Judd-Ofelt theory. The lifetime of (3)P(0) and (3)P(1) levels has been measured. Fluorescence quenching has been observed for higher concentrations of Pr(3+) ion. The temperature dependence of the fluorescence intensity and the lifetime of the (3)P(0) level has been investigated and found that they decrease with the increase of the temperature. PMID- 16257730 TI - Modern spectroscopic techniques in the characterization of Schiff base macrocyclic ligand and its complexes with transition metals. AB - Mn(II), Co(II), Ni(II), and Cu(II) complexes with a new azamacrocyclic tetradentate [N(4)] ligand i.e. 2,3,9,10-tetraphenyl;l,4,8,11 tetraazacyclotetradeca;1,3,8,10-tetraene (L) have been synthesized and characterized by elemental analysis, molar conductance measurements, magnetic susceptibility measurements, mass, (1)HNMR, IR, electronic and EPR spectral studies. On the basis of their non-electrolytic nature, the probable formula of the complexes is proposed to be [M(L)X(2)], where M=Mn(II), Co(II), Ni(II), and Cu(II), X=Cl(-) and NO(3)(-), in dimethylformamide (DMF). All the complexes are of high-spin type and found to have six coordinated, octahedral geometry for Mn(II), Co(II), and Ni(II) complexes, and tetragonal for Cu(II) complexes. Macrocyclic ligand and its complexes have also been screened against pathogenic bacteria and fungi in vitro as growth inhibiting agent. PMID- 16257731 TI - Preparation of a novel fluorescence nanoparticles and its application in the determination of Hg(II). AB - The strong fluorescence 2-vinylnaphthalene and acrylic acid polymer nanoparticles have been prepared under ultrasonic radiation. Based on the fluorescence quenching of polymer by Hg(II), a method for the selective determination of Hg(II) was developed. The reaction conditions between Hg(II) and polymer were investigated in detail. The assay is very few interference stable fluorescence signals (at least 2h), simple instrument (common spectrofluorometer) and simple step. Under optimal experimental conditions, a limit of detection of 0.01 microg ml(-1) was achieved. The calibration curve was linear over the concentration range 0.04-0.1 microg ml(-1) with a correlation coefficient of 0.9927. The proposed method has been applied to the selective quantification of Hg(II) in synthetic samples with the satisfactory results. PMID- 16257732 TI - Spectra and structure of binary azeotropes I. Acetone-chloroform. AB - Some characteristic vibrational modes of acetone and chloroform change due to the azeotrope formation. The extend of interaction of these molecules has significant effects on some vibrational modes involved, depending on unit structure in azeotrope cluster. Besides (1)H NMR signals undergo some chemical shifts, which show the effect of oncoming molecules on the target molecule. FT-IR and (1)H NMR spectra of pure substances and corresponding azeotrope were recorded, mutual influences due to azeotrope formation based on mole ratio, boiling point and spectral changes has been discussed. Unit structure of cluster deduced by investigating fundamental frequency shifts and (1)H NMR chemical shifts. PMID- 16257733 TI - Preparation, characterization and biological activity of Fe(III), Fe(II), Co(II), Ni(II), Cu(II), Zn(II), Cd(II) and UO(2)(II) complexes of new cyclodiphosph(V)azane of sulfaguanidine. AB - Novel hexachlorocyclodiphosph(V)azane of sulfaguanidine, H(4)L, l,3-[N'-amidino sulfanilamide]-2,2,2,4,4,4-hexachlorocyclodiphosph(V)azane was prepared and its coordination behaviour towards the transition metal ions Fe(III), Fe(II), Co(II), Ni(II), Cu(II), Zn(II), Cd(II) and UO(2)(II) was studied. The structures of the isolated products are proposed based on elemental analyses, IR, UV-vis, (1)H NMR, mass spectra, reflectance, magnetic susceptibility measurements and thermogravimetric analysis (TGA). The hyperfine interactions in the isolated complex compounds were studied using 14.4keV gamma-ray from radioactive (57)Co (Mossbauer spectroscopy). The data show that the ligand are coordinated to the metal ions via the sulfonamide O and deprotonated NH atoms in an octahedral manner. The H(4)L ligand forms complexes of the general formulae [(MX(z))(2)(H(2)L)H(2)O)(n)] and [(FeSO(4))(2) (H(4)L) (H(2)O)(4)], where X=NO(3) in case of UO(2)(II) and Cl in case of Fe(III), Co(II), Ni(II), Cu(II), Zn(II) and Cd(II). The molar conductance data show that the complexes are non electrolytes. The thermal behaviour of the complexes was studied and different thermodynamic parameters were calculated using Coats-Redfern method. Most of the prepared complexes showed high bactericidal activity and some of the complexes show more activity compared with the ligand and standards. PMID- 16257734 TI - The viscosity and temperature dependence of (1)H T(1)-NMRD of the Gd(H(2)O)(8)(3+) complex. AB - Water proton T(1)-NMRD profiles of the Gd(H(2)O)(8)(3+) complex have been recorded at three temperatures and at four concentrations of glycerol. The analysis is performed using both the generalized Solomon-Bloembergen-Morgan (GSBM) theory, and the stochastic Liouville approach (SLA). The GSBM approach uses a two processes dynamic model of the zero-field splitting (ZFS) correlation function whereas SLA uses a single process model. Both models reproduce the proton T(1)-NMRD profiles well. However, the model parameters extracted from the two analyses, yield different ESR X-band spectra which moreover do not reproduce the experimental ESR spectra. It is shown that the analyses of the proton T(1) NMRD profiles recorded for a solution Gd(H(2)O)(8)(3+) ions are relatively insensitive to the slow modulation part of dynamic model of the ZFS interaction correlation function. The description of the electron spin system results in a very small static ZFS, while recent ESR lineshape analysis indicates that the contribution from the static ZFS is important. Analysis of proton T(1)-NMRD profiles of Gd(H(2)O)(8)(3+) complex do result in a description of the electron spin system but these microscopic parameters are uncertain unless they also are tested in a ESR-lineshape analysis. PMID- 16257735 TI - Density functional theory study of the Fourier transform infrared and Raman spectra of Cu(II) bis-acetylacetone. AB - Fourier transform infrared and Fourier transform Raman spectra of Cu(II) bis acetylacetone have been obtained. The geometry, frequency and intensity of the vibrational bands of this compound and its 1,5-(13)C(2), 3-(13)C, 1,3,5-(13)C(3), 2,4-(13)C(2), (18)O(2) and 2,4-(13)C(2)-(18)O(2) derivatives were obtained by the density functional theory (DFT) with the B3LYP functional and using the 6-31G(*) and 3-21G(*) basis sets. The calculated frequencies are compared with the solid infrared and Raman spectra. All the measured infrared and Raman bands were interpreted in terms of the calculated vibrational modes. The percentage of deviation of the bond lengths and bond angles gives a good picture of the normal modes, and serves as a basis for the assignment of the wavenumbers. Most computed bands are predicted to be at higher wavenumbers than the experimental bands. The calculated geometrical parameters show slight differences compared with the experimental results. These differences can be explained by the different physical state of Cu(II) bis-acetylacetone. The DFT-B3LYP calculations assumed a free molecule in the gas phase. Analysis of the vibrational spectra indicates a strong coupling between the chelated ring modes. PMID- 16257736 TI - Conformational analysis and crystal structure of {[1-(3-chloro-4-fluorobenzoyl)-4 fluoropiperidin-4yl]methyl}[(5-methylpyridin-2-yl)methyl]amine, fumaric acid salt. AB - {[1-(3-Chloro-4-fluorobenzoyl)-4-fluoropiperidin-4yl]methyl}[(5-methylpyridin-2 yl)methyl]amine, fumaric acid salt (C(20)H(22)ClF(2)N(3)O, C(4)H(4)O(4)) (1) was synthesized and characterized by the complete (1)H, (13)C and (19)F NMR analyses. The conformation of the piperidin ring, in the solution state, was particularly studied from the coupling constants determined by recording a double-quantum filtered COSY experiment in phase-sensitive mode. (1)H NMR line-shape analysis was used, at temperatures varying between -5 and +60 degrees C, to determine the enthalpy of activation of the rotational barrier around the CN bond. Compound 1 crystallizes in the triclinic space group P1 with a=8.517(3) Angstrom, b=12.384(2) Angstrom, c=12.472(3) Angstrom, alpha=70.88(2) degrees, beta=82.04(2) degrees, gamma=83.58(2) degrees. The results strongly indicate that the solid and solution conformations are similar. Thermal stability and phases transitions were investigated by thermal gravimetric analysis (TGA) and differential scanning calorimetry (DSC). Furthermore polymorphism screening was studied from recrystallization of 1 performed in seven solvents and by slurry conversion in water. The X-ray powder diffraction (XRPD) and differential scanning calorimetry results suggested that 1 crystallizes into one crystalline form which melts at 157 degrees C (DeltaH=132 J g(-1)). PMID- 16257737 TI - Spectral analysis of Cu(2+): B(2)O(3)--ZnO--PbO glasses. AB - A new series of heavy metal oxide (PbO) based zinc borate glasses in the chemical composition of (95-x)B(2)O(3)-5ZnO-xPbO (x=10, 15, 20, 25, 30, 35, 40, 45 and 50 mol%) have been prepared to verify their UV filtering performance. Both direct and indirect optical band gaps (E(opt)) have been evaluated for these glasses. For a reference glass of 45B(2)O(3)-5ZnO-50PbO, refractive indices at different wavelengths are measured and found the results satisfactorily correlated with the theoretical data upon the computation of Cauchy's constants of A=1.766029949, B=159531.024 nm(2) and C=-1.078 x 10(10) nm(4). Measurements concerning X-ray diffraction (XRD), FT-IR, differential scanning colorimeter (DSC) profiles have been carried out for this glass. The FT-IR profile has revealed that the glass has both BO(3) and BO(4) units. From DSC thermogram, glass transition temperature (T(g)), crystallization temperature (T(c)) and melting temperature (T(m)) have been located and from them, other related parameters of the glass have also been calculated. Visible absorption spectra of 45B(2)O(3)-5ZnO-(50-x)PbO-xCuO (x=0. 1, 0.2, 0.5 and 1.0 mol%) have revealed two absorption bands at around 400 nm ((2)B(1g)-->(2)E(g)) and 780 nm ((2)B(1g)-->(2)B(2g)) of Cu(2+) ions, respectively. Emission bands at 422 and 512 nm are found for the 1 mol % CuO doped glass with excitations at 306 and 332 nm. PMID- 16257738 TI - Spectrofluorometric study of complexation of some amino derivatives of 9,10 anthraquinone with beta-cyclodextrin. AB - Complexation reactions between 1-amino-9,10-anthraquinone (AA1), 1-amino-2-methyl 9,10-anthraquinone (AA2), 1-amino-2,4-dimethyl-9,10-anthraquinone (AA3) and 1 amino-2-ethyl-9,10-anthraquinone (AA4) and beta-cyclodextrin were studied spectrofluorometrically, under optimized experimental conditions. The formation constants of the resulting 1:1 beta-cyclodextrin complexes were evaluated and found to decrease in the order AA4>AA1>AA3>AA2. Possible reasons for the observed stability sequence are discussed based on the structures proposed for the resulting inclusion complexes. PMID- 16257739 TI - Synthesis and spectroscopic characterization of some transition metal complexes of a new hexadentate N(2)S(2)O(2) Schiff base ligand. AB - A novel interesting hexadentate dibasic N(2)S(2)O(2) donor Schiff base ligand, H(4)dcsalpte, was synthesized by the condensation of 3-formylsalicylic acid and 1,2-di(o-aminophenylthio)ethane and characterized. The reactions of the ligand with different metal(II/III)salts under varied reaction conditions afforded a series of metal complexes. The ligand, H(4)dcsalpte, behaves either as a dibasic or neutral hexadentate one, depending on the reaction conditions. Structural investigations on the ligand and their complexes have been made based on elemental analyses, molar conductance values, magnetic moment values, cryomagnetic and spectral (UV-vis, IR, (1)H NMR, and Mossbauer) data. Based on magnetic susceptibility, Mossbauer and electronic spectral data the iron(III) complex [Fe(III)(H(2)dcsalpte)]ClO(4) (8), isolated in the present investigation, it is inferred that the spin states 5/2 and 1/2 are in equilibrium. Similarly a tri-iron(III) complex [Fe(III)(3)(H(2)dcsalpte)(H(3)dcsalpte)Cl(3)]Cl(3) (7), isolated in this study, has been inferred to contain two iron(III) sites in tetrahedral environment and one in the octahedral environment. The aerial oxidation of an equimolar mixture of H(4)dcsalpte and Co(CH(3)COO)(2).4H(2)O in ethanol under reflux gave two products, [Co(H(2)dcsalpte)]CH(3)COO (10) and [(Hbtcsaldm)Co(Hbvcsaldm)] (11), a cobalt(III) complex bound to two dissimilar tridentate NSO donor ligands formed as a result of the oxidative cleavage of the CS bond. In the complex 11, Hbtcsaldm stands for the dianion of the tridentate Schiff base ligand N-(2'-benzenethiol)-3-carboxysalicylaldimine and Hbvcsaldm stands for the mono anion of the tridentate Schiff base ligand N-(benzene-2'-S vinyl)-3-carboxysalicylaldimine, both being formed as a result of the oxidative cleavage of H(4)dcsalpte. PMID- 16257740 TI - The control of phthalocyanine properties through nitro-group electronic effect. AB - The UV-vis spectra of peripherally substituted tetranitrometallophthalocyanines (TNMPcs) 1a-1d and non-peripherally substituted TNMPcs 2a-2d were investigated. In comparison of 1a with 2a, there is the only difference in the substitution position of nitro-groups. The structural diversity of 1a and 2a resulted in different electronic effects of nitro-group on Pc rings, which caused them have two kinds of Q bands. The Q band in the UV-vis spectra of 1a-1d was split into two peaks, which were obviously influenced by temperature, while no split was observed in the Q band of 2a-2d. We can draw a conclusion that tuning nitro-group electronic effect is an important approach for controlling the properties of Pcs. PMID- 16257741 TI - Use of 1,3-diaminepropane-3-propyl grafted onto a silica gel as a sorbent for flow-injection spectrophotometric determination of copper (II) in digests of biological materials and natural waters. AB - The 1,3-diaminepropane-3-propyl-anchored silica gel (DAPPS) was successfully employed as a sorbent in a spectrophotometric flow system for the preconcentration of Cu(2+) in digests of biological materials (maize powder, soybean, citrus leaves, corn stalks) as well as water samples (river, stream, streamlet, springwater and well). The system presented a minicolumn packed with DAPPS, where the sample solution was passed through it for a period of time, and subsequently, an eluent solution, stripped-out the retained analyte which was further determined with DDTC at 460 nm. The better preconcentration conditions utilized were: 120s loading, 60s elution, 30s regeneration of the column, loading flow rate 6.5 ml min(-1), buffer solution for the preconcentration and regeneration of the column-borate buffer pH 8.5, elution flow rate 2.3 ml min( 1), time of elution 60s, eluent composition, 0.4 mol l(-1) HNO(3). Under these conditions, the preconcentration factor obtained was 36, and the detection limit achieved was 8.4 ng ml(-1) in water samples and 0.84 microg g(-1) in biological material. The maximum adsorption capacity of DAPPS to Cu(2+) was 0.49 mmol g(-1) (31.1 mg g(-1)) obtained in a batch system. The recovery of copper in the water samples ranged from 96.9 to 102.4% and in the biological materials ranged from 97.0 to 102.6%. PMID- 16257742 TI - Characterization of southern Taiwan red soils as a regenerable sorbent for sorption of hydrogen sulfide from coal gas with spectroscopic techniques. AB - The purpose of this research is to use red soils as a high-temperature regenerable sorbent for sorption of hydrogen sulfide from coal gas and collocates with a series of spectroscopic apparatuses in order to investigate the structure changes after multiple sorption/regeneration processes. Results indicate that red soils could be reused after 10 multiple sorption/regeneration cycles and maintain approximately 80% sorption efficiency. With EDS and EA analyses, residual sulfur species are detected in the 10th regenerated red soils and exist with values of 0.6 and 0.33%, respectively. Undesired sulfur species including sulfide, sulfate and elemental sulfur are further identified by XPS spectroscopy. With the best regression fitting results, sulfate species is the dominative sulfur species, which occupies approximately 71% of residual sulfur. It is believed that these residual sulfur species are the major cause to result in red soils' deterioration after regeneration and reduce the sorption efficiency. Appreciable amounts of regeneration gases CO, CO(2) and SO(2) are detected by on-line FTIR spectroscopy. Their formation mechanisms are attributed to the different gas-solid reaction, one is the reaction of carbon and oxygen, and the other is FeS and oxygen. From the analysis of solid-state nuclear magnetic resonance spectroscopy (SSNMR), a significant change for (27)Al and (29)Si and it is established that a portion of aluminum is reformed into pentacoordinated structure. Formation of aluminosilicate is maybe another reason to result in deterioration as a result of the loss of surface area. PMID- 16257743 TI - The monothiocyanate complexes of chromium ion(III) on the silver electrode by the surface enhanced Raman scattering. AB - Two adsorbate forms of the monothiocyanate complex of chromium ion on the silver electrode are identified in the surface enhanced Raman scattering. The spectroscopic, especially the electronic, properties of these two forms under different applied voltages on the electrode and under both 632.8 and 514.5 nm excitations are studied by the bond force constants (bond orders) and the bond polarizability derivatives which are retrieved from the Raman intensities by an algorithm developed by Wu and co-workers. The work shows the potential of this approach to the surface enhanced Raman scattering and other fields like resonance Raman that involve vibronic coupling. PMID- 16257744 TI - UV-vis spectroscopy for following the kinetics of homogeneous polymerization of diphenylamine in p-toluene sulphonic acid. AB - Kinetics of chemical oxidative polymerization of diphenylamine (DPA) was followed in aqueous 1M para-toluene sulphonic acid (p-TSA) using potassium peroxomonosulphate (PMS) or peroxodisulphate (PDS), independently as an oxidant. The medium was found to be homogeneous and became dark green in colour during the course of polymerisation. The course of polymerization was followed by UV-vis spectroscopy. Rate of polymerization (R(p)) was determined for various conditions by following the absorbance values corresponding to poly(diphenylamine) (PDPA) for different concentrations of DPA and PMS or PDS at various time intervals of polymerization. The observed dependences of DPA, PDS or PMS on R(p) were used to deduce rate equations for PDS or PMS initiated polymerization of DPA. The rate constant for the formation of poly(diphenylamine), was estimated. In situ spectroelectrochemical studies on the polymerization of DPA were also carried out on an ITO electrode in 1M p-TSA. The results are in accordance with the intermediates suggested in chemical oxidative polymerization. PMID- 16257745 TI - The spectroscopic properties of Er(3+)-doped TeO(2)-Nb(2)O(5) glasses with high mechanical strength performance. AB - (100-x)TeO(2)-xNb(2)O(5) (x=5-20) niobic tellurite glasses doped with 0.5 mol.% Er(2)O(3) were synthesized, and their thermal, mechanical, and spectroscopic properties were measured and compared to the properties of the typical 75TeO(2) 20ZnO-5Na(2)O (TZN) tellurite glass. The refractive index (n(d)), density (rho), and glass transition temperature (T(g)) of bulk glasses increase with the Nb(2)O(5) content. The Vickers microhardness (H(v)) of bulk glass in niobic tellurite glasses also increases with the Nb(2)O(5) content. The values (2.5 3.2GPa) of H(v) in the niobic tellurite glasses are 47-88% larger than that (1.7GPa) in TZN glass. The effect of Nb(2)O(5) content on absorption spectra, the Judd-Ofelt parameters Omega(t) (t=2, 4, 6), fluorescence spectra and the lifetimes of Er(3+):I(13/2) level were also investigated, and the stimulated emission cross-section was calculated from McCumber theory. With increasing Nb(2)O(5) content in the glass composition, the Omega(t) (t=2, 4, 6) parameters, fluorescence full width at half maximum (FWHM) of I(13/2) of Er(3+) increase, while the (4)I(13/2) lifetimes of Er(3+) decreases. Compared with TZN glass, the gain bandwidth properties of Er(3+)-doped TeO(2)-Nb(2)O(5) glass is much larger than in tellurite glass based TeO(2)-ZnO-Na(2)O system, bismush-based glass, germanate, and silicate glasses, which indicates that TeO(2)-Nb(2)O(5) glasses are better choice as a practical available host material for broadband Er(3+) doped amplifier. PMID- 16257746 TI - Depolarization ratio and correlation between the relative intensity data and the abundance ratio of various isotopes of liquid carbon tetrachloride at room temperature. AB - The room temperature Stokes and anti-Stokes Raman spectra of liquid CCl(4) have been recorded. The intensity ratios of anti-Stokes to Stokes Raman bands as a function of Raman shift are obtained in agreement with polarizability theory. The depolarization ratio rho (nu) as a function of Raman shift is obtained also in agreement with automatically scanned depolarization ratio rho (nu). Ratio of the intensity of the isotopic nu(1) bands indicates small deviation from the theoretical relative abundance of CCl(4) isotopes. The intensity ratio of the [nu(3)-nu(4), (nu(1)+nu(4))-nu(4)] and nu(1) bands is obtained. The consequences of the presence of different isotopes of CCl(4) on the depolarization ratio of its vibrational bands are discussed. The effects of impurities in liquid CCl(4) on depolarization ratio of the nu(1) band are estimated. PMID- 16257747 TI - Spectrophotometric study of the reaction mechanism between DDQ Pi- and iodine sigma-acceptors and chloroquine and pyrimethamine drugs and their determination in pure and in dosage forms. AB - Two simple and accurate spectrophotometric methods are presented for the determination of anti-malarial drugs, chloroquine phosphate (CQP) and pyrimethamine (PYM), in pure and in different pharmaceutical preparations. The charge transphere (CT) reactions between CQP and PYM as electron donors and 2,3 dichloro-5,6-dicyano-p-benzoquinone (DDQ) pi-acceptor and iodine sigma-acceptor reagents to give highly coloured complex species have been spectrophotometrically studied. The optimum experimental conditions have been studied carefully. Beer' law is obeyed over the concentration range of 1.0-15 microg ml(-1) for CQP and 1.0-40 microg ml(-1) for PYM using I(2) and at 5.0-53 microg ml(-1) for CQP and 1.0-46 microg ml(-1) for PYM using DDQ reagents, respectively. For more accurate results, Ringbom optimum concentration range is calculated and found to be 10-53 and 8-46 microg ml(-1) for CQP and PYM using DDQ, respectively and 5-15 and 8-40 microg ml(-1) for CQP and PYM using iodine, respectively. The Sandell sensitivity is found to be 0.038 and 0.046 g cm(-2) for DDQ method and 0.0078 and 0.056 g cm( 2) for I(2) method for CQP and PYM, respectively which indicates the high sensitivity of both methods. Standard deviation (S.D.=0.012-0.014 and 0.013 0.015) and relative standard deviation (R.S.D.=0.09-1.4 and 1.3-1.5%) (n=5) for DDQ and I(2) methods respectively, refer to the high accuracy and precision of the proposed methods. These results are also confirmed by between day precision of percent recovery of 99-100.6%, and 98-101% for CQP and PYM by DDQ method and 99-102% and 99.2-101.4% for CQP and PYM by I(2) method respectively. These data are comparable to those obtained by British and American pharmacopoeias assay for the determination of CQP and PYM in raw materials and in pharmaceutical preparations. PMID- 16257748 TI - Quantitative determinations of levofloxacin and rifampicin in pharmaceutical and urine samples using nuclear magnetic resonance spectroscopy. AB - Rapid, specific and simple methods for determining levofloxacin and rifampicin antibiotic drugs in pharmaceutical and human urine samples were developed. The methods are based on (1)H NMR spectroscopy using maleic acid as an internal standard and DMSO-d6 as NMR solvent. Integration of NMR signals at 8.9 and 8.2 ppm were, respectively, used for calculating the concentration of levofloxacin and rifampicin drugs per unit dose. Maleic acid signal at 6.2 ppm was used as the reference signal. Recoveries of (97.0-99.4)+/-0.5 and (98.3-99.7)+/-1.08% were obtained for pure levofloxacin and rifampicin, respectively. Corresponding recoveries of 98.5-100.3 and 96.8-100.0 were, respectively, obtained in pharmaceutical capsules and urine samples. Relative standard deviations (R.S.D.) values < or =2.7 were obtained for analyzed drugs in pure, pharmaceutical and urine samples. Statistical Student's t-test gave t-values < or =2.87 indicating insignificant difference between the real and the experimental values at the 95% confidence level. F-test revealed insignificant difference in precisions between the developed NMR methods and each of fluorimetric and HPLC methods for analyzing levofloxacin and rifampicin. PMID- 16257749 TI - Vibrational dephasing and hydrodynamic effects on vibrational relaxation rates in acetophenone: Raman bandshape analysis. AB - The isotropic component of Raman band for C=O stretching mode of acetophenone in solution was analyzed by estimating the correlation coefficient with reference to Lorentzian lineshape. In the intermediate region of solute/solvent concentration there is a sharp decrease in the correlation coefficient and there appears to be a transition from non-Lorentzian to Lorentzian lineshape. The vibrational relaxation rates have been estimated from the isotropic component of Raman band in different solvents. The rate is shown to be dependent on several macroscopic as well as microscopic properties of the solute-solvent system and intermolecular interactions. The hydrodynamic and dispersion forces appear to play a major role in determining the vibrational relaxation rate and the broadening of the bands. PMID- 16257750 TI - Spectroscopic properties of neodymium(III)-containing polyoxometalates in aqueous solution. AB - The spectroscopic properties of the neodymium(III)-containing polyoxometalates (POMs) [Nd(PW(11)O(39))(2)](11-), [Nd(PMo(2)W(9)O(39))(2)](11-), [Nd(PMo(4)W(7)O(39))(2)](11-), [Nd(PMo(6)W(5)O(39))(2)](11-), [Nd(SiMo(2)W(9)O(39))(2)](13-), [Nd(P(2)W(17)O(61))(2)](17-), [NdW(10)O(36)](9-), [NdP(5)W(30)O(110)](12-) and [NdAs(4)W(40)O(140)](25-) are described. Absorption spectra of aqueous solutions of the complexes have been recorded and the transition intensities are parameterised in terms of the Judd-Ofelt intensity parameters Omega(lambda) (lambda=2, 4, 6). Marked differences were found between the luminescence lifetimes of the complexes of the type Nd(POM) and those of the type Nd(POM)(2), due to a better shielding of the neodymium(III) ions from the bulk water molecules in the latter type of complexes. PMID- 16257751 TI - The hydrogen bonding and conformations of p-tert-butylcalix[4]arene as studied by IR spectroscopy and by DFT calculations. AB - IR and far IR spectra of p-tert-butylcalix[4]arene were recorded at various temperatures between 16 and 180 degrees C and spectra of solutions and crystalline solids were obtained. Ab initio density functional calculations gave vibrational frequencies and infrared intensities for four conformers: cone, partial cone, 1,2- and 1,3-alternate. Complete assignments were made for experimental IR spectra of the cone conformer. The bands characteristic for each conformation were defined. It was revealed that O--H stretching low-frequency shift Deltanu in the cone conformation exceeds Deltanu shifts for other conformers. The effect was stipulated by a cooperative interaction of cyclic hydrogen bonds. The obtained spectra-structure correlation can be used for characteristic of calixarenes conformation. PMID- 16257752 TI - Identification of static JT in copper(II) doped hexaimidazole M(II) lattices: M=Co and Ni: an EPR study. AB - Single crystal EPR studies on Cu(II) doped paramagnetic host lattices, hexaimidazole M(II) dichloride tetrahydrate (M=Co and Ni), isomorphous with M=Zn, have been carried out from room temperature to 77K to understand the nature of Jahn-Teller (JT) distortion in these paramagnetic host systems. The paramagnetic impurity, doped in the present two paramagnetic host lattices, shows anisotropic EPR spectra with superhyperfine from ligands, even at room temperature. An interesting observation noticed in the EPR spectra at room temperature is that there are more resonances corresponding to the second site in the paramagnetic hosts than in the diamagnetic host at 4.2K. This difference in behavior between the diamagnetic and paramagnetic host lattices indicates a change in the depth of the JT valleys. The spin Hamiltonian parameters are evaluated for Cu(II) ion in both the host lattices and the relaxation times have been calculated for the ion in cobalt host lattice only. PMID- 16257753 TI - FT-Raman spectroscopic evidences for the preferential solvation of sodium tetrafluorobrate in acetonitrile-based mixed solvents. AB - Solutions of sodium tetrafluorobrate in acetonitrile-dimethylformamide, and acetonitrile-dimethylsulfoxide mixtures have been studied by FT-Raman spectroscopy for three solvent compositions, respectively. New bands due to solvent molecules in the first solvation shell of Na(+) were detected in the region of the O=C--N deformation and CH(3) rocking mode for amide and of the S=O and C--S stretching modes for sulfoxide. The individual solvation numbers of sodium cation in different environment were deduced. In all the cases, it is found that the sodium ion was preferentially solvated by DMF or DMSO in respective binary solvents. This result was further supported by ab initio calculations. PMID- 16257754 TI - Complex formation between aliphatic amines and chromogenic calix[4]arene derivatives studied by FT-IR spectroscopy. AB - Complex formation of two calix[4]arene derivatives - containing two ethoxycarbonylmethoxy groups or a bridge including two carboxamide moieties in the coordination sphere - with aliphatic amines (n-octylamine, OA and triethylamine, TEA) were the subject of FT-IR studies. Two nitrophenyl type auxochromic moieties were also present in both of the ligands. Solvent effects were studied in chloroform, diiodomethane and dichloromethane. Presumably, in these reactions, the reactants undergo phenol-amine type interactions and, simultaneously, coordinative interaction takes place between the carbonyl group of the ligand and the protonated form of the aliphatic amine. Complex formation of calixarene 1a with n-octylamine was complete resulting in drastic changes in the FT-IR spectra. Moreover, the possibility of the formation of a new compound by the reaction between the ester type functionalities and octylamine was precluded. The expected ligand was synthesized and its spectra were compared with those of calixarene 1a. In most of the cases of calixarene 1b, the amide moiety of the carboxamide bridge does not participate in the complex formation with the exception of complexes formed in diiodomethane solution and of the 1b-TEA complex observed in dichloromethane. PMID- 16257755 TI - Spectroscopic studies, cyclic voltammetry and synthesis of nickel(II) complexes with N(4), N(2)O(2) and N(4)S(2) donor macrocyclic ligands. AB - Nickel(II) complexes of the general composition Ni(L)X(2) (where X=SCN, NO(3) and 1/2SO(4) and ligands=L(1) L(2) and L(3)) have been synthesized and characterized by elemental analyses, magnetic moments, IR, (1)H NMR, (13)C NMR and electronic spectral studies. Nickel(II) ions, such as nitrates, thiocyantes and sulphates were found to act as templates for the cyclic condensations [1+1] and [2+2] of NH(2--)C(6)H(4)--O--CH(2)--CH(2)--O--C(6)H(4)--NH(2), NH(2)--(CH(2))(2)--NH(2) and NH(2)--CH(CH(3))--CH(2)--NH(2) with C(6)H(5)--CO--CO--C(6)H(5), C(6)H(5)--CO- CH(2)--CO--C(6)H(5) and (COOH--CH(2)--CH(2))(2)S. All the complexes show magnetic moments corresponding to two unpaired electrons except [Ni(L(1))](NO(3))(2) and [Ni(L(2))](NO(3))(2) complexes which are diamagnetic. Electronic spectroscopy was used to analyse the differences between the paramagnetic and diamagnetic forms. Electrochemical properties have been studied extensively for Ni(III/II) and Ni(II/I) couples. The equilibrium between the paramagnetic and diamagnetic forms and the nickel(III/II) couple are strongly dependent on the electrolyte. It has been observed that the sulphate group coordinated selectively on the apical position of the nickel(II) centers of the compounds. The structural and electrochemical studies suggest that cooperative effects, involving coordination of sulphate to one nickel center, is responsible for the recognition of this anion. Various ligand field parameters have been calculated and discussed. PMID- 16257756 TI - Spectrophotometric and electrical studies of charge transfer complexes of 2-amino 1,3,4-thiadiazole with pi-acceptors. AB - Charge transfer (CT) complexes formed between 2-amino-1,3,4-thiadiazole as donor and 2,3-dichloro-5,6-dicyano-p-benzoquinone (DDQ), p-chloranil (p-CHL), o chloranil (o-CHL), p-bromanil (BRL) and chloranilic acid (CHA) as acceptors, have been studied spectrophotometrically. Benesi-Hildebrand and Job continuous variation methods were applied to the determination of association constant (K), molar extinction coefficients (epsilon), dipole moment and stoichiometric ratio, respectively. The solid CT complexes have been synthesized and characterized by different spectral methods. The spectral changes reveal that the CT interaction depends on the type of the acceptors. The magnetic properties of the various complexes were also investigated. The electrical properties for the solid CT complexes are measured from which the activation energies are calculated. PMID- 16257757 TI - ATR-FTIR spectroscopy as a way to identify natural protein-based materials, tortoiseshell and horn, from their protein-based imitation, galalith. AB - This paper presents a new totally non-destructive methodology, for the identification of protein-based materials, tortoiseshell, horn and galalith, on heritage objects. Attenuated total reflexion (ATR) infrared spectroscopy combined to a deconvolution procedure of amide I band was performed to characterize the secondary structure of these materials and allows us to identify each material. The component at 1639 cm(-1), attributed to the amide group involved in hydrated random-coil domains, allows us to distinguish horn from galalith and tortoiseshell. A second component, at 1614 cm(-1), assigned to amide group involved in protein self-association, allows us to differentiate galalith and tortoiseshell. This differentiation is applicable to both raw and manufactured materials. Then, we applied our models to identify what unknown object was made of. PMID- 16257758 TI - EPR, UV-vis, magnetic, spectral studies and electrochemical behaviour of mononuclear transition metal complexes derived from novel hexa-aza-macrotricyclic ligand. AB - Aza-macrocyclic complexes have gained importance because of their pharmacological properties. Hexa-aza-macrocycles containing glutarimide efficiently coordinate as hexa-dentate ligand, to give complexes of Cu(II) possessing tetragonal structure and Mn(II), Co(II) and Ni(II) metal ions that are essentially octahedral. Spectroscopic, and chemical characterizations of these systems are presented in this article. For Ni(II) complexes results on electron transfer processes measured by cyclic voltammetry and colourimetry have been studied. PMID- 16257759 TI - The investigation of chemical structure of coal macerals via transmitted-light FT IR microspectroscopy. AB - The Late Permian coals from South China are characterized by high content of the unique maceral "barkinite" and elemental hydrogen, typically produce high yields of tar, and exhibit significant oil-producing potential. "Barkinite" has been identified as suberinite for a long time, but now many Chinese workers have concluded that "barkinite" is not suberinite, rather it is a distinct maceral. The term "barkinite" was formally certified and named by the State Bureau of Technical Supervision of PR China in 1991, 1995, and 1999, however, it has not been recognized as a scientific term by international coal geologists and the ICCP. Transmitted-light FT-IR microspectroscopy and curve-fitting analysis were used to investigate the chemical structure of "barkinite"; at the same time, parallel studies were also carried out on vitrinite, fusinite, and sporinite. In comparison with other maceral types, the micro-FT-IR spectra of "barkinite" are characterized by very strong intensities of aliphatic CH(x) stretching vibrations at 3000-2800 cm(-1) and deformations at 1460-1450 cm(-1), less intense bands from aromatic CC ring stretching at 1610-1600 cm(-1) and aromatic CH out-of-plane deformations at 900-700 cm(-1). The aliphatic side-chains in the molecular structure of "barkinite" are longer and less branched. In addition, there also appear intense aliphatic ether COC and alcohol CO stretching bands at 1100-1000 cm(-1), notable aromatic CH stretching vibrations at 3050-3030 cm(-1), weak OH stretching bands of water in the 3600-3200 cm(-1) range, and rare acid CO group absorption at 1740-1700 cm(-1). Collectively the IR spectral characteristics indicate that "barkinite" is composed of great numbers of long chain aliphatics, a fewer of aromatics and rare of oxygenate components. The chemical structure of "barkinite" show that the high tar yields and the relatively high oil-producing potential of the Late Permian coals from South China are attributable to the high "barkinite" content in the Leping coals and the great numbers of long chain aliphatic substances contained in this maceral. PMID- 16257760 TI - Preparation and application of a novel core/shell organic nanoparticle as a fluorescence probe in the selective determination of Cr(VI). AB - A novel core/shell organic nanoparticles, anthracene/poly-acrylamide (AN/PAM), has been prepared successfully. Based on the fluorescence quenching of AN/PAM nanoparticles by Cr(VI), a method for the selective determination of Cr(VI), without separation of Cr(VI) in water, was developed. Furthermore, the reaction mechanism between nano-AN/PAM and Cr(VI) was also discussed. The synthesis and reaction conditions were investigated in detail. The assay is characterized by short reaction time, very few interference stable fluorescence signals, simple instruments and sensitivity. Under optical experimental conditions, a limit of detection of 0.02 microg/ml was achieved. The calibration curve was linear over the concentration range 0.04-2.00 microg/ml with a correlation coefficient of 0.9924. The proposed method has been applied to the selective quantification of Cr(VI) in synthetic samples and waste-water samples with the satisfactory results. PMID- 16257762 TI - Fluorescence rejection in Raman spectra of Syncrude Sweet Blend distillation fractions. AB - Four techniques for the reduction or elimination of fluorescence from Raman spectra of Syncrude process samples were examined in this study. These methods are based on the retrieval of Raman bands from differential, or derivative spectra. Differential data were generated by subtracting similar spectra of a given sample obtained in three ways: (a) shifted detection utilizing an array detector and two successive spectrometer settings; (b) shifted excitation (dispersive Raman) where the two spectra are recorded using neighbouring laser lines and ordinary photon counting; (c) shifted excitation (FT-Raman) in which the laser frequency is changed in software before acquisition of the second spectrum. In addition to these differential techniques, derivative spectra were acquired directly with a dispersive Raman system by modulating the wavelength during scanning. These fluorescence rejection methods were applied to two groups of Syncrude Sweet Blend distillation fractions. For light gas oils (boiling range, 195-343 degrees C) the ratio of monocyclic and bicyclic aromatic species was determined and bands due to aliphatic CH(n) groups were characterized. Heavy gas oils (343-524 degrees C) yielded bands that allowed quantitation of monocyclic, bicyclic and total aromatic groups. Bands due to aliphatics were also identified for the heavy gas oils. These results constitute a significant advance compared to the information obtainable using conventional dispersive and FT-Raman spectroscopy for the analysis of hydrocarbon distillation fractions. PMID- 16257761 TI - Scaled quantum chemical studies of the structure and vibrational spectra of 2 (methylthio) benzimidazole. AB - The solid phase FT-IR and FT-Raman spectra of 2-(methylthio) benzimidazole (2 MTBI) have been recorded in the regions 4000-50 cm(-1) and 3500-100 cm(-1), respectively. The spectra were interpreted with the aid of normal coordinate analysis following full structure optimization and force field calculations based on density functional theory (DFT) using standard B3LYP/6-31G* and B3LYP/6 311+G** methods and basis set combinations and was scaled using various scale factors yielding fairly good agreement between observed and calculated frequencies. Based on the present good quality scaled quantum mechanical (SQM) force field, a reliable description of the fundamentals was provided and the assignments have been proposed with the aid of normal coordinate analysis. The infrared and Raman spectra were also predicted from the calculated intensities. Comparison of the simulated spectra with the experimental spectra provides important information about the ability of the computational method to describe the vibrational modes. PMID- 16257763 TI - Photophysical processes of some benzimidazole derivatives. AB - The photophysical properties of N-(alpha-naphthyl)-benzimidazole (alpha-NABI), N (beta-naphthyl)-benzimidazole (beta-NABI) and N-(alpha-pyridyl)-benzimidazole (alpha-PYBI) were studied and alpha-NYBI exhibit intramolecular charge transfer fluorescence in polar solvents. The fluorescence of benzimidazoles can be quenched by acetic acid and the existence of exciplexes was observed between the benzimidazole derivatives and acetic acid. Particularly, the maximum emission peak of solution of alpha-PYBI in mixed solvent, ether and acetic acid, presents obvious red-shift with the increase of concentration of acetic acid in the mixed solvent. PMID- 16257764 TI - Electronic states of Ga(3)Si, GaSi(3), and their ions. AB - The equilibrium geometries and electronic states of Ga(3)Si, GaSi(3), and their ions are investigated using the complete active space self-consistent-field (CASSCF) and DFT(B3LYP)/CCSD(T) techniques. The (2)B(1), (3)B(1), and (1)A(1) states in C(2v) symmetry with a planar quadrilateral geometry are found to be the ground states of Ga(3)Si, Ga(3)Si(+), and Ga(3)Si(-), respectively. On the other hand, the ground states of GaSi(3), GaSi(3)(-) are also predicted to undergo Jahn Teller distortion to the (2)A'' and (1)A' states in C(s) with a distorted triangular pyramid geometry, respectively, whereas that of GaSi(3)(+) is found to be the (1)A(1) state in C(3v) with symmetric triangular pyramid structure. Binding energies, electron affinities, ionization energies of Ga(3)Si and GaSi(3) are computed at the CCSD(T)/QCISD(T) level and discussed. PMID- 16257765 TI - (1)H, (13)C NMR and X-ray crystallographic studies of highly polyhalogenated derivatives of costunolide lactone. AB - The costunolide lactone, a sesquiterpene compound isolated from Zaluzania triiloba species, reacted with several dihalocarbene sources produced by trihaloform-NaOH under successive phase transfer reactions yielding mono-, bis- and tris-dihalocyclopropane adducts. The structures, as well as the configurational assignments of the different derivatives, were established by (1)H and (13)C NMR spectroscopy and assisted by X-ray crystallographic and molecular modelling studies. The specific shielding of protons in the neighbourhood of different halogens on the cyclopropane moieties was correlated to the pseudocontact interactions. PMID- 16257766 TI - Enhanced sensitivity of lithium dithionates doped with rhodium and nickel for EPR dosimetry. AB - Electron paramagnetic resonance (EPR) studies of X-irradiated lithium dithionate, Li(2)S(2)O(6) x 2H(2)O, doped with Ni and Rh have shown that these impurities enhance the yield of radicals formed by X-irradiation at room temperature. The signal in the doped samples, measured peak-to-peak of the single EPR derivative line attributed to the SO(3)(-) anion was about 3-4 times that of the pure lithium dithionate and more than 10 times stronger than the alanine signal. These impurities also shortened the spin-lattice relaxation time, T(1), which gives the possibility to measure the doped samples at a higher microwave power. This implies that sensitivity could be further enhanced in the already sensitive EPR dosimeter material lithium dithionate. PMID- 16257767 TI - UV-vis, IR and (1)H NMR spectroscopic studies of some Schiff bases derivatives of 4-aminoantipyrine. AB - Five Schiff bases derived from 4-aminoantipyrine and benzaldehyde derivatives (I) are prepared and their UV-vis, IR, (1)H NMR and fluorescence spectra are investigated and discussed. The electronic absorption spectra of the hydroxy 4 aminoantipyrine Schiff bases Ib and Ie as well as the fluorescence spectra of Ie are studied in the organic solvents of different polarity. The UV-vis absorption spectra of 4-aminoantipyrine Schiff bases Ib, Id and Ie are investigated in aqueous buffer solutions of varying pH and utilized for the determination of pK(a) and DeltaG of the ionization process. The reactions of the hydroxy compounds Ib and Ie with Ni(II) and Cu(II) ions are also studied. The results of spectral studies are supported by some molecular orbital calculations using an atom superposition and electron delocalization molecular orbital theory for a compound Ib. PMID- 16257768 TI - Infrared and Raman spectroscopic studies of l-valine l-valinium perchlorate monohydrate. AB - FT-IR and FT-Raman spectra were recorded and analyzed for l-valine l-valinium perchlorate monohydrate crystals. The wave number assignments have been made for the functional groups, viz. COOH, COO(-), --[NH(3)](+), C--(CH(3))(2), C--C--N and C--H. One of the two amino acid residues remains in the zwitterionic form while the other residue exists in the cationic form. The symmetry of the ClO(4)( ) anion has been found to be lowered corresponding to ClO(2) group. The hydrogen bonds that prevail between amino acid residues, perchlorate anion and water molecule influence the wave numbers of several stretching and deformation modes to deviate from the expected values. PMID- 16257769 TI - Determination of trace aluminum by fluorescence quenching method based on catalysis of potassium chlorate oxidizing alizarin red. AB - A new method for the determination of trace aluminum has been proposed. It is based on the fact that alizarin red can emit strong and stable fluorescence at 80 degrees C for 30 min and Al(3+) can effectively catalyze potassium chlorate oxidizing alizarin red to form non-fluorescence complex which cause the fluorescence quenching. The linear dynamic range of this method is 0.040-4.00 ngl(-1) with a detection limit of 5.3 pgl(-1). The regression equation can be expressed as DeltaI(f)=8.731+21.73c(Al(3+)) (ngl(-1)), with the correlation coefficient r=0.9992 (n=6). This sensitive, rapid and accurate method has been applied to the determination of trace aluminum(III) in human hair and tea samples successfully. What is more, the mechanism of catalyzing potassium chlorate oxidizing alizarin red by the fluorescence quenching method is also discussed. PMID- 16257771 TI - Optical spectroscopy of Yb(3)Al(5)O(12) single crystal. AB - Yb(3)Al(5)O(12) single crystal has been grown by Czochralski (CZ) method. The absorption spectrum was investigated at low temperature and the electronic energy levels for (2)F(5/2) multiplet of Yb(3+) in YbAG was proposed. The up-conversion emission of the crystal under 940 nm diode pumping and the X-ray excited luminescence (XEL) features of the crystal were also studied. PMID- 16257770 TI - Spectral studies on the interaction between lanthanum ion and the ligand: N,N' ethylenebis-[2-(o-hydroxyphenolic)glycine]. AB - The interaction between N,N'-ethylenebis-[2-(o-hydroxyphenolic)glycine] (EHPG) and lanthanum was studied by the difference UV spectra and fluorescence spectra. At pH 7.4, 0.01 M N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid (Hepes), with the addition of 1.0 x 10(-3)M lanthanum, two new peaks were observed at 238 nm and 294 nm by absorptivity spectroscopy compared with blank solution EHPG suggesting the interaction of lanthanum and EHPG. At the same time, the reaction could be measured by fluorescence spectra. The fluorescence intensity of EHPG at 310 nm was significantly decreased in the presence of lanthanum. The 1:1 stoichiometric ratio of EHPG to lanthanum was confirmed by both fluorescence and UV titration curves. In addition, the molar absorptivity of La-EHPG at 238 nm is (1.23+/-0.01)x10(4)cm(-1)M(-1). The conditional binding constant was calculated to be log K(La-EHPG)=12.09+/-0.37 on the basis of the result of UV titration curves. PMID- 16257772 TI - Thermodynamics study of the dimerization equilibria of rhodamine B and 6G in different ionic strengths by photometric titration and chemometrics method. AB - The dimerization constants of rhodamine B and 6G have been determined by studying the dependence of their absorption spectra on the temperature in the range 20-80 degrees C at different total concentrations of rhodamine B (5.89 x 10(-6) to 2.36 x 10(-4)M) and rhodamine 6G (2.34 x 10(-5) to 5.89 x 10(-4)M) and in different concentrations of LiCl, NaCl and KCl salts as supporting electrolytes. The monomer-dimer equilibrium of rhodamine B and 6G have been determined by chemometrics refinement of the absorption spectra obtained by thermometric titrations performed at different ionic strengths. The quantitative analysis of the data of undefined mixtures, was carried out by simultaneous resolution of the overlapping spectral bands in the whole set of absorption spectra. The dimerization constants are varied by changing the ionic strength and the degree of dimerization are decreased by increasing of the ionic strength of the medium. The enthalpy and entropy of the dimerization reactions were determined from the dependence of the equilibrium constants on the temperature (van't Hoff equation). From the thermodynamic results the TDeltaS degrees -DeltaH degrees plot was sketched. It shows a fairly good positive correlation which indicates the enthalpy-entropy compensation in the dimerization reactions (compensation effect). PMID- 16257773 TI - Determination of the overlapping pK(a) values of chrysin using UV-vis spectroscopy and ab initio methods. AB - The overlapping pK(a) values of 5,7-dihydroxyflavone (chrysin) in EtOH-water solutions were determined by means of a UV-vis spectroscopic method that uses absorbance diagrams, at constant ionic strength (0.050 M) and temperature (25.0+/ 0.1 degrees C). It was observed that the pK(a) values increase when the polarity polarizability and solvation abilities of the reaction medium decrease. In order to calculate the pK(a1) and pK(a2) of chrysin in pure water, various relationships between the determined pK(a) and properties of solvents (relative permittivity, alpha-parameter of Taft and parameter Acity), are proposed. Moreover, with the aim of explaining the first pK(a1) value obtained, the molecular conformations and solute-solvent interactions of the 7(O(-))chrysinate monoanion were also investigated, using ab initio methods. Several ionization reactions and equilibria in water, which possesses a high hydrogen-bond-donor ability, are proposed. These reactions and equilibria constituted the necessary theoretical basis to calculate the first acidity constant of chrysin. The HF/6 31G(d) and HF/6-31+G(d) methods were used for calculations. Tomasi's method was used to analyze the formation of intermolecular hydrogen bonds between the 7(O( ))chrysinate monoanion and water molecules. It was proposed that in alkaline aqueous solutions the monoanion of chrysin is solvated with one water molecule. The agreement between the experimental and theoretical pK(a1) values provides good support for the acid-base reactions proposed in this paper. PMID- 16257774 TI - Electronic and infrared vibrational analysis of cyanidin-quercetin copigment complex. AB - Copigment complex formation between cyanidin and quercetin, in aqueous buffered solutions, was studied by electronic absorption and infrared vibrational spectroscopies. It was found that the association of cyanidin with quercetin occurred at pH 3.0 and pH 5.0 including cyanidin flavylium ion and anhydrobase transformation forms, respectively. Obtained copigmentation constant values of K=2726.7 (pH 3.0) and K=1093.1 (pH 5.0) indicated good association ability of the investigated molecules. Infrared spectra revealed the existence of hydrogen bonds in the copigment complexes structures. The analysis of the deconvoluted infrared spectra indicated several types of hydrogen bonds, differently formed: the H- O...H bonds with the corresponding bands around 3500 cm(-1) and bonds formed via H(3)O(+), oxonium, ion of the molecules with the corresponding bands below 3000 cm(-1). PMID- 16257775 TI - An experimental and theoretical vibrational spectroscopic study of [AsPh(4)](2)[Sn(dmit)(3)] x Me(2)CO. AB - As shown previously by X-ray structure determinations, [tris(1,3-dithiole-2 thione-4,5-dithiolato)stannate(IV)](2-) salts, [Q](2)[Sn(dmit)(3)], contain isolated cations and dianions. While the tin centres generally having octahedral geometries, the overall shapes of the dianions of these complexes in the solid state can differ with conformations varying from T, Y to asymmetrical arrangements. We now report, as a follow up to our earlier study on the Y-shaped complex, [NEt(4)](2)[Sn(dmit)(3)], an experimental and theoretical study of the vibrational spectra of solid solvated {[AsPh(4)](2)[Sn(dmit)(3)] x Me(2)CO}, in which the dianion has a T-shaped conformation. The infrared and Raman spectra, recorded from 4000 to 150 cm(-1), have been analysed by different ab initio calculations based on restricted Hartree-Fock (RHF) and density functional theory (DFT-Beck3LYP). The calculations were carried out on isolated dianions and cations with the 6-31G and 6-31G(d) basis sets and effective core potentials of Steven, Bash and Krauss (SBK). Fundamentals, overtones and combinations have been assigned. Generally, the Y- and T-shaped dianions exhibit similar infrared/Raman spectra, apart from differences in the C=C and the symmetrical M-S stretching frequencies: such differences can be used diagnostically to distinguish the overall shape of the tris(chelated)metallate dianion. PMID- 16257776 TI - Upconversion luminescence and mechanisms of Tm(3+)/Yb(3+)-codoped oxyhalide tellurite glasses. AB - To obtain efficient blue upconversion laser glasses, upconversion luminescence and mechanisms of Tm(3+)/Yb(3+)-codoped oxyhalide tellurite glasses were investigated under 980 nm excitation. The results showed that upconversion blue and red emission intensities of Tm(3+) first increase, reach its maximum at Tm(2)O(3)%=0.1 mol %, and then decrease with increasing Tm(2)O(3) content. The effect of Tm(2)O(3) content on upconversion intensity is discussed, and possible effect mechanisms are evaluated. The investigated results were conducing to increase upconversion luminescence efficiency of Tm(3+). PMID- 16257777 TI - Metal isotope and density functional study of the tetracarboxylatodicopper(II) core vibrations. AB - Vibrational spectra of tetrakis(acetato)diaquadicopper(II) complex have been deeply examined in order to provide a detailed description of dynamics of [Cu(2)O(8)C(4)] core being a typical structural unit of most copper(II) carboxylates. Low frequency bands related to significant motions of metal atoms were detected by metal isotope substitution. Observed spectra and isotope shifts were reproduced in DFT calculations. For clear presentation of computed normal vibrations, a D(4h) symmetry approximation was successfully applied. Basing on observed isotope shifts and calculation results, all skeletal vibrations have been analyzed including normal mode with the largest Cu...Cu stretching amplitude assigned to Raman band at 178 cm(-1). PMID- 16257778 TI - Preparation of fluorescent polyvinyl alcohol keto-derivatives nanoparticles and selective determination of chromium(VI). AB - A novel fluorescent polyvinyl alcohol keto-derivatives nanoparticle (PVAK) has been prepared in one-step method. The nanoparticles has excitation and emission maxima at 349 and 462 nm, respectively. Based on the fluorescence quenching of PVAK by Cr(VI), we established a simple and selective fluorimetric method for the determination of Cr(VI) without separation of Cr(III) in water. The reaction conditions between Cr(VI) and PVAK were investigated in detail. Furthermore, the reaction mechanism between PVAK and Cr(VI) was also discussed. Under optimal experimental conditions, a limit of detection of 0.02 microg mL(-1) was achieved. The calibration curve was linear over the concentration range 0.1-13.2 microg mL( 1) with a correlation coefficient of 0.9987. The proposed method has been applied to the selective quantification of Cr(VI) in synthetic samples and waste-water samples with the satisfactory results. PMID- 16257779 TI - Studies on the riboflavin-resorcinol interaction in an aqueous medium and its pH dependence. AB - The equilibrium constants, K', for the formation of the molecular association complex between riboflavin and resorcinol in the pH range 6-8 were found to be in the range 5-25 mol(-1)dm(3) by difference absorption spectroscopy. The equilibrium constants from emission spectroscopy were estimated to be in the range 38-55 mol(-1)dm(3) over the same pH range. Both sets of K' show a remarkable dependence on pH. The equilibrium constants K(A) and K(B) describing the interaction of riboflavin with neutral and ionic forms of resorcinol, respectively, obtained by resolving K', shows that K(B) is much greater than K(A). Molecular modeling studies suggest a stacked conformation of the two components in the complexed form. The interaction energies obtained from modeling studies also suggest a stronger interaction between the ionic form of resorcinol and riboflavin. PMID- 16257780 TI - Solution NMR studies of supramolecular complexes of [60]- and [70]fullerenes with mono O-substituted calix[6]arene. AB - Supramolecular complexation of [60]- and [70]fullerenes with 37-allyl 38,39,40,41,42-pentahydroxy-5,11,17,23,29,35-hexa(4-tert butyl)calix[6]arene (I) has been studied in CCl(4) medium by NMR spectrometric method. All of the complexes are found to be stable with 1:1 stoichiometry. Formation constants (K) of the above supramolecular complexes have been determined from systematic variation of NMR chemical shifts of specific protons of I in the presence of [60] and [70]fullerenes. Trends in the K value suggest that [70]fullerene binds more strongly with I relative to [60]fullerene. Both PM3 and ab initio calculations reveal that the intermolecular interaction in the [70]fullerene/I complex proceeds through quite deep energy minima. PMID- 16257781 TI - Interaction of phenazine with water and DNA bases. AB - The fluorescence spectrum of aqueous phenazine (PZ), an N-heterocyclic compound, shows some interesting features that indicate the formation of PZ-water complex in the excited state. Two types of complexes are postulated; Type I, formed by the association of water molecule with one of the nitrogen of PZ and Type II, formed by the association of water molecules with both the nitrogen of PZ. In addition, PZ also interacts with the DNA bases, adenine and thymine and the corresponding nucleosides, adenosine and thymidine. Fluorescence and laser flash photolysis studies indicate that the mode of interaction may be photoinduced electron transfer. PMID- 16257782 TI - FTIR, FT-Raman spectra and ab initio DFT vibrational analysis of 2-bromo-4-methyl phenylamine. AB - The FTIR and FT-Raman spectra of 2-bromo-4-methyl-phenylamine (BMP) have been recorded. From the standard geometrical parameters the geometry of BMP was optimized at ab initio and DFT levels of theory with complete relaxation in the potential energy surface using 6-311+g(d,p) and 6-311+g(2df,2p) basis sets. Several thermodynamic parameters were also calculated for the minimum energy conformer at ab initio and DFT level of theories. The harmonic vibrational frequencies were calculated and the scaled values have been compared with experimental FTIR and FT-Raman spectra. Majority of the computed wavenumbers were found to be in good agreement with the experimental observations. The experimental spectra also coincide satisfactorily with those of theoretically constructed bar type spectrograms. PMID- 16257784 TI - Keynote comment: not just an exercise in solidarity. PMID- 16257783 TI - Bridging the gap. PMID- 16257785 TI - Role of neoadjuvant androgen deprivation before radical prostate radiotherapy. PMID- 16257786 TI - Fever of unknown origin and cancer: a clinically meaningful association. PMID- 16257787 TI - Prediction of chemotherapy-induced anaemia: is knowledge really power? PMID- 16257788 TI - Neoadjuvant chemoradiotherapy in resectable oesophageal cancer. PMID- 16257790 TI - Boost to cancer care in India. PMID- 16257791 TI - Short-term androgen deprivation and radiotherapy for locally advanced prostate cancer: results from the Trans-Tasman Radiation Oncology Group 96.01 randomised controlled trial. AB - BACKGROUND: Androgen deprivation is an established treatment regimen for disseminated prostate cancer; however, its role in patients with localised cancer is less clear. We did a large randomised controlled trial to determine whether 3 months or 6 months of androgen deprivation given before and during radiotherapy improves outcomes for patients with locally advanced prostate cancer. METHODS: 818 men with locally advanced prostate cancer were randomly assigned to: no androgen deprivation (ie, radiotherapy alone: 66 Gy in 33 fractions of 2 Gy per day over 6.5-7.0 weeks to the prostate and seminal vesicles); 3 months' androgen deprivation with 3.6 mg goserelin given subcutaneously every month and 250 mg flutamide given orally three times a day starting 2 months before radiotherapy (same regimen as control group); or 6 months' androgen deprivation, with the same regimen, starting 5 months before radiotherapy (same regimen as control group). Primary endpoints were time to local failure and prostate-cancer-specific survival; secondary endpoints were distant failure, disease-free survival, and freedom from salvage treatment. Analyses were done by intention to treat. FINDINGS: 802 (98%) patients were eligible for analysis. Median follow-up was 5.9 years (range 0.1-8.5). Compared with patients assigned no androgen deprivation, those assigned 3 months' treatment had significantly improved local failure (hazard ratio [HR] 0.56 [95% CI 0.39-0.79], p=0.001), biochemical failure-free survival (0.70 [0.56-0.88], p=0.002), disease-free survival (0.65 [0.52-0.80], p=0.0001), and freedom from salvage treatment (0.73 [0.56-0.96], p=0.025). 6 months' androgen deprivation significantly improved local failure (0.42 [0.28 0.62], p<0.0001), biochemical failure-free survival (0.58 [0.46-0.74], p<0.0001), disease-free survival (0.56 [0.45-0.69], p<0.0001), freedom from salvage treatment (0.53 [0.40-0.71], p<0.0001), distant failure (0.67 [0.45-0.99], p=0.046) and prostate-cancer-specific survival (0.56 [0.32-0.98], p=0.04) compared with no androgen deprivation. INTERPRETATION: 6 months' androgen deprivation given before and during radiotherapy improves the outlook of patients with locally advanced prostate cancer. Further follow-up is needed to estimate precisely the size of survival benefits. Increased radiation doses and additional periods of androgen deprivation might lead to further benefit. PMID- 16257792 TI - Fever of unknown origin and cancer: a population-based study. AB - BACKGROUND: Fever of unknown origin is associated with cancer, but the tumour types most commonly involved and the prognostic implications of this relation have not been studied. We aimed to assess the risk of cancer and survival after hospital admission for fever of unknown origin in a nationwide Danish follow-up study. METHODS: We linked the Danish national registry of patients and the nationwide cancer registry, and assessed cancer risk for 43,205 patients discharged for the first time from Danish hospitals after admission for fever of unknown origin during 1977-97. We compared cancer frequency with that expected on the basis of cancer incidence in the general population, and survival of patients with cancer who had had fever of unknown origin with that of controls (ie, patients with cancer matched for cancer site, age at time of cancer diagnosis, and year of cancer diagnosis). FINDINGS: Median follow-up was 6.3 years (IQR 2.7 12.3). During the first year of follow-up, 399 cancers were diagnosed in those who had had fever of unknown origin (standardised incidence ratio 2.3 [95% CI 2.1 2.5]). These individuals had an increased risk of developing cancer--in particular haematological malignant disease; sarcoma; and cancers of the liver, brain, kidney, colon, and pancreas. After 1 year of follow-up, individuals had an increased risk of developing haematological malignant disease and cancers of the liver, brain, and kidney. A slightly higher proportion of people diagnosed with cancer within 1 year of hospital discharge after fever of unknown origin had distant metastases compared with controls (101 [34%] of 294 vs 819 [28%] of 2907; prevalence ratio 1.2 [95% CI 1.0-1.4], and had worse survival than did controls (mortality rate ratio 1.4 [95% CI 1.2-1.6]). INTERPRETATION: Hospital admission for fever of unknown origin is a marker of occult cancer and is associated with a poor outlook. PMID- 16257793 TI - Chemotherapy-induced anaemia during adjuvant treatment for breast cancer: development of a prediction model. AB - BACKGROUND: At present, oncologists prescribe chemotherapy according to standard dose schedules, and as a result many patients develop serious, dose-limiting toxic effects such as anaemia. We aimed to develop a prediction model for anaemia in patients with breast cancer who were receiving adjuvant chemotherapy. METHODS: We reviewed medical records of 331 patients who had received adjuvant chemotherapy for breast cancer. Patients were divided randomly into a derivation sample (n=221) and internal-validation sample (n=110). An external sample of 119 patients enrolled onto the control group of a randomised trial of epoetin alfa was used to validate the model further. Multivariable logistic regression was applied to develop the initial model. We then developed a risk-scoring system, ranging from 0 (low risk) to 50 (high risk), based on the final regression variables. A receiver operating characteristic (ROC) curve analysis was done to measure the accuracy of the scoring system when applied to both validation samples. FINDINGS: The risk of anaemia increased as the pretreatment haemoglobin concentration decreased and was reduced with successive chemotherapy cycles. Risk was also predicted by a platelet count of 200x10(9) cells/L or less before chemotherapy, age 65 years or older, type of adjuvant chemotherapy, and use of prophylactic antibiotics. ROC analysis had acceptable areas under the curve of 0.88 for the internal-validation sample and 0.84 for the external validation sample. A risk score of > or = 24 to < 25 before chemotherapy was identified as the optimum cut-off for maximum sensitivity (83.5%) and specificity (92.3%) of the prediction model. INTERPRETATION: The application and continued refinement of this prediction model will help oncologists to identify patients at risk of developing anaemia during chemotherapy for breast cancer, and might enhance patient-centred care by the application of anaemia treatment in a proactive and appropriate way. PMID- 16257794 TI - Survival after surgery for oesophageal cancer: a population-based study. AB - BACKGROUND: Oesophageal resection alone remains the only potentially curative treatment for oesophageal cancer. However, few population-based studies of survival after surgery have been done. We aimed to assess whether survival after surgery for oesophageal cancer has improved since 1987. METHODS: We identified from the nationwide cancer register 764 patients in Sweden who had undergone resection alone for oesophageal cancer between Jan 1, 1987 and Dec 31, 2000, and followed up these individuals until Oct 18, 2004, through linkage to the nationwide registers of death, emigration, and total population. Tumour stage, location, and histology were recorded by review of histopathological reports. Hazard ratios, with 95% CI, for death adjusted for age; sex; comorbidity; and tumour stage, location, and histology were calculated by Cox proportional-hazards regression model. FINDINGS: Overall, the proportion of deaths up to 30 days after surgery decreased from 10.1% in 1987-91 to 4.9% in 1997-2000. Increased long-term survival was associated with low tumour stage. Survival was 46.5% 1 year after surgery, 24.1% 3 years after surgery, and 19.7% 5 years after surgery in 1987-91; 58.4%, 31.7%, and 24.9%, respectively, in 1992-96; and 61.7%, 39.9%, and 30.7%, respectively, in 1997-2000. The adjusted hazard ratio for death was 0.76 (95% CI 0.62-0.92) in 1992-96 and 0.57 (0.45-0.71) in 1997-2000. INTERPRETATION: Survival after surgery for oesophageal cancer has improved substantially since 1987. Because this improvement cannot be explained by a change in patient characteristics or tumour characteristics, it is probably a result of better surgery. PMID- 16257795 TI - Increasing incidence of colorectal cancer in Asia: implications for screening. AB - Many Asian countries, including China, Japan, South Korea, and Singapore, have experienced an increase of two to four times in the incidence of colorectal cancer during the past few decades. The rising trend in incidence and mortality from colorectal cancer is more striking in affluent than in poorer societies and differs substantially among ethnic groups. Although changes in dietary habits and lifestyle are believed to be the reasons underlying the increase, the interaction between these factors and genetic characteristics of the Asian populations might also have a pivotal role. Non-polypoidal (flat or depressed) lesions and colorectal neoplasms arising without preceding adenoma (de novo cancers) seem to be more common in Asian than in other populations. The absence of polypoid growth preceding malignancy has posed difficulties in screening for early colorectal cancer by radiological imaging or even endoscopic techniques. Although epidemiological data are scanty, most Asian populations are not aware of the growing problem of colorectal cancer. More work is needed to elucidate the magnitude of the problem in Asia. PMID- 16257796 TI - Lymphatic mapping in management of patients with primary cutaneous melanoma. AB - In patients with primary cutaneous melanoma, knowledge of regional lymph-node status provides important information on outlook. Evidence suggests that early removal of nodes that contain metastatic disease improves survival outcome. Lymphatic drainage occurs first to sentinel nodes, which are therefore the nodes most likely to contain metastatic disease. Lymphatic mapping with lymphoscintigraphy is important to identify reliably sentinel nodes for removal and thus establish the status of regional nodes. Mapping studies in patients with melanoma have provided new insights into lymphatic anatomy and have shown previously unsuspected drainage pathways, which have important implications for accurate identification and removal of sentinel nodes. Because it is impossible to predict the site or sites of sentinel nodes clinically in individual patients, routine preoperative lymphoscintigraphy is a prerequisite if reliable results are to be obtained from sentinel-node biopsy. PMID- 16257797 TI - Selection of adjuvant chemotherapy for treatment of node-positive breast cancer. AB - Over the past two decades, several studies have suggested that regimens that contain anthracyclines are more effective than those that do not. A meta-analysis by the 2005 Early Breast Cancer Trialists' Collaborative Group confirmed that about 6 months of anthracycline-based polychemotherapy in the adjuvant setting reduced the yearly death rate from breast cancer by about 38% for women younger than 50 years and by 20% for women aged 50-69 years. Although this meta-analysis found that survival was better with regimens that contain anthracycline than with regimens based on cyclophosphamide, methotrexate, and fluorouracil, the best use of anthracycline-based regimens remains unclear. Adjuvant regimens in use can be categorised into three groups: standard-dose anthracycline; escalated-dose epirubicin; and anthracyclines and taxanes. The duration of treatment and combination of dose and drugs varies between these three categories. We reviewed the three types of regimen to establish which provide a better outcome in terms of safety, efficacy, cost, and convenience to patients. We found that both escalated-dose epirubicin and anthracycline-taxane regimens were most effective in terms of disease-free survival and overall survival. Of the specific anthracycline-based regimens, the docetaxel, doxorubicin, and cyclophosphamide regimen (TAC); the fluorouracil, 100 mg epirubicin, and cyclophosphamide regimen (FEC100); and the cyclophosphamide, epirubicin, and fluorouracil regimen (CEF) produced the greatest proportional decreases in 5-year death rate. PMID- 16257798 TI - Use of cancer chemopreventive phytochemicals as antineoplastic agents. AB - A lot of information has been gathered on cellular mechanisms by which chemopreventive phytochemicals, such as curcumin (a spice in curry) or epigallocatechin gallate (extracted from tea), interfere with carcinogenesis. A comparison of this knowledge with what we know about molecularly targeted chemotherapeutic agents suggests that it might be worthwhile to investigate the usefulness of such phytochemicals in the treatment of established malignant diseases. Phytochemicals use a plethora of antisurvival mechanisms, boost the host's anti-inflammatory defence, and sensitise malignant cells to cytotoxic agents. The restricted systemic availability of agents such as curcumin and epigallocatechin gallate, needs to be taken into account if they are to be developed as cochemotherapeutic drugs. PMID- 16257799 TI - Isolated bilateral adrenal choriocarcinoma. PMID- 16257800 TI - Complete resolution of anal cancer after chemotherapy for acute myeloid leukaemia. PMID- 16257801 TI - Acute arterial occlusion after chemotherapy for testicular cancer. PMID- 16257802 TI - Spirituality among young women at risk for eating disorders. AB - OBJECTIVE: This study examined the spiritual and religious (S/R) beliefs and practices of college-age women at high-risk for eating disorders, and the relationship between body image distress, coping, and S/R. METHOD: Two hundred fifty-five college-age women with elevated weight and shape concerns, assessed using the Weight/Shape Concerns Scale and the Eating Disorder Examination (EDE), completed surveys about their S/R beliefs and practices. RESULTS: Women with strong S/R beliefs and practices cope with body dissatisfaction differently than women without strong S/R beliefs. Participants with strong S/R were significantly more likely to pray, meditate, or read religious/spiritual texts to cope with body image distress. Participants without strong beliefs and practices were more likely to cope utilizing distraction. Women with strong beliefs who prayed found it effective. DISCUSSION: Study participants were heterogenenous in their S/R beliefs and practices. These beliefs and practices may be underutilized resources for coping with body image concerns. PMID- 16257804 TI - Negative emotion and disordered eating among obese college students. AB - The present study examined the relationship between negative emotions, body dissatisfaction, exercise, and disordered eating attitudes and behaviors among obese college students. It also examined whether there were gender differences in these variables. A total of 88 males and 102 females, who reported a BMI score above 30, completed a survey. Females reported higher levels of disordered eating, body dissatisfaction, and more frequent dieting than males and as predicted, males reported higher levels of exercise behaviors. Body dissatisfaction, anger discomfort, and self dissatisfaction all correlated with drive for thinness for both genders. Anger discomfort was the only variable to predict disordered eating for both genders. The results support numerous studies that have found that females are at greater risk of disordered eating than males, and also suggest that anger management may be an important component in treatment of disordered eating among obese young adults. PMID- 16257803 TI - Worry and eating disorders: a psychopathological association. AB - Worry is a mental process associated with anxiety disorders. The key feature of worry is the predominance of a negative-type and preoccupied thought about possible threatening future events. OBJECTIVE: Some studies have shown that worry may be a feature of eating disorders. This study aims to measure whether worry is significantly higher in eating disordered individuals than in a normal control group and whether worry is associated with the Eating Disorders Inventory. METHODS: Sixty-three individuals affected by an eating disorder (34 anorexics and 29 bulimics) completed the Penn State Worry Questionnaire, the Structured Clinical Interview for DSM, and the Eating Disorder Inventory. Thirty normal controls completed the Penn State Worry Questionnaire. RESULTS: Penn State Worry Questionnaire scores were significantly higher in eating disordered individuals than in controls. It was associated with all the symptoms of eating disorders and was correlated with all the EDI subscales, except for the subscale 'bulimia'. These findings suggest that worry is important for understanding the psychopathology of eating disorders. PMID- 16257805 TI - Mediators of the association between abuse and disordered eating in undergraduate men. AB - The vast majority of ED research has focused on women. However, recent studies have suggested that ED symptomatology in men may be underestimated. Additional investigations are needed to better understand EDs and their correlates among men. This study examined the relationships between childhood abuse experiences and disordered eating in male undergraduates. In addition, potential mediators and moderators of the association between abuse and disordered eating were evaluated. Results indicated that physical abuse and physical neglect were the only adverse childhood experiences associated with disordered eating. In addition, depression mediated the associations between these forms of abuse and ED symptomatology. However, neither anxiety nor alexithymia were significant mediators of the association between abuse and EDs. Social support moderated the association between physical neglect and depression, such that individuals with high social support were less depressed than those with low social support, regardless of their level of physical neglect. These results are somewhat different than those found in exclusively female samples, highlighting the importance of specifically examining EDs and their correlates among men. PMID- 16257806 TI - Ethnic and gender differences in eating attitudes among black and white college students. AB - This study examines ethnic and gender differences in eating attitudes and behaviors among college students. Participants were 225 Black and 199 White students sampled from a historically Black university. White participants were more dissatisfied with their bodies, engaged in more self-loathing, and dieted more than Blacks. Similarly, women were more dissatisfied with their bodies, engaged in more self-loathing, dieted more, and showed a greater drive for thinness than men. White women and Blacks of either gender exhibited similar predictors of drive for thinness with each group showing some combination of dieting and self-loathing. Intrapersonal anger predicted drive for thinness in White men, adding to a growing body of research suggesting a link between anger and eating disorders. Results support a substantial body of literature showing that Black and White college students differ on their views of body image and eating. Future research should explore the role of anger as a risk factor for eating disorders among White men. PMID- 16257807 TI - Gender differences regarding psychopathological, family and social characteristics in adolescents with abnormal eating behavior. AB - OBJECTIVE: (1) To identify gender differences regarding psychopathological, family and social characteristics in adolescents with abnormal eating behavior; (2) to study risk factors for this abnormal eating in boys and girls. METHOD: Adolescents participating in this community-based longitudinal study completed, at the age of 13 (t(1)) and 15 years (t(2)), a semi-structured interview and the validated Spanish versions of several self-reported questionnaires measuring eating and general psychopathology. A control group of 150 pupils and 159 adolescents with abnormal eating behavior were selected. RESULTS: Girls with abnormal eating showed significantly more psychiatric morbidity and boys more social difficulties. Body dissatisfaction and psychiatric morbidity predicted abnormal eating at t(2) only in girls. DISCUSSION: The findings contribute to the debate on gender differences in abnormal eating behavior etiology. PMID- 16257808 TI - Assessing the functional nature of binge eating in the eating disorders. AB - This study explored the functional nature of binge eating through the development of a new self-report instrument called the Binge Eating Adjective Checklist. Participants were 405 adult females who presented to a specialized eating disorders clinic. A subset of participants with bulimia nervosa also completed additional psychometrics and treatment. Those participants who reported greater reductions in negative affective and somatic states during a binge episode were also more distressed on measures of perfectionism, self-esteem, ineffectiveness, and interceptive awareness. They were also less likely to achieve abstinence from bingeing and vomiting over the course of treatment. Thus, the instrument appears to offer a methodology for studying the phenomenology of binge eating and the prediction of therapeutic outcome in bulimia nervosa. PMID- 16257809 TI - Parental rearing behaviours and eating disorders: the moderating role of core beliefs. AB - OBJECTIVE: Core beliefs have been shown to mediate between eating psychopathology and dysfunctional parent-daughter interactions. However, the possible moderating role of core beliefs has been neglected. This study aimed to explore the hypothesis that core beliefs serve as moderator variables in the relationship between recalled parental rearing behaviours and eating psychopathology. METHOD: Sixty-six women with a current eating disorder completed self-report measures of parental rearing behaviours, core beliefs, and eating psychopathology. RESULTS: Three core beliefs were found to moderate the relationship between paternal rejection and aspects of eating psychopathology. The predictive validity of paternal rejection on aspects of eating symptomatology was found to decrease as dysfunctional core beliefs increased. DISCUSSION: When levels of social isolation, vulnerability to harm, and self-sacrifice core beliefs were high, recalled parental relationships were no longer relevant to current eating psychopathology. The findings provide further evidence that core beliefs are important factors in eating disorder psychopathology and may be clinically useful in identifying targets for treatment. PMID- 16257810 TI - Consistency of food intake over four eating sessions in the laboratory. AB - OBJECTIVE: The aim of this study was to test the consistency (repeatability) of laboratory measures of food intake, including eating microstructure (cumulative food intake curves), in non-obese humans across four eating sessions, each separated by 1 week. A secondary aim was to test the effect of dietary restraint on the food intake of female participants. METHODS: Mixed model analyses were used to compare average food intake across sessions, and intraclass correlation coefficients (ICC) were employed to compare within subject variability to total variance. High and low restraint females and low restraint males consumed four lunches in a Universal Eating Monitor (UEM) laboratory. The lunches consisted of one type of sandwich (chicken salad sandwich squares) or three types of sandwiches (chicken salad, ham, and turkey sandwich squares) presented in counterbalanced order. RESULTS: Measures of food intake were stable for men and women, regardless of sandwich variety. In females, level of dietary restraint (high vs. low) did not significantly affect food intake. DISCUSSION: The results indicate that eating behavior in the laboratory is sufficiently stable over time to justify evaluation of interventions designed to alter food intake using within subject designs. PMID- 16257811 TI - The Minnesota Eating Behavior Survey: a brief measure of disordered eating attitudes and behaviors. AB - This article describes details of the development and psychometric characteristics of a brief self-report inventory for assessing attitudes and behaviors symptomatic of eating disorders that is currently in use in a longitudinal study of over 700 families with 11-year-old or 17-year-old twin girls. The Minnesota Eating Behavior Survey (MEBS), formerly the Minnesota Eating Disorder Inventory, is a 30-item measure developed for use with children as young as 10 years as well as adults. An examination of the MEBS's psychometric properties in a large, community sample of girls, women, and men demonstrated good factor congruence, internal consistency reliability, three-year stability, as well as evidence of concurrent and criterion validity. This questionnaire has promise as a screening and assessment measure for eating disturbance in cross sectional and longitudinal research involving individuals of a wide range of ages. PMID- 16257812 TI - Construct validity and reliability of the College Oriented Eating Disorders Screen (COEDS). AB - The College Oriented Eating Disorders Screen [COEDS; Nowak, J.A., Roberson-Nay, R., Strong, D.R., Bucceri, J. and Lejuez, C.W. (2003). Using item response theory in the development and validation of the College-Oriented Eating Disorders Screen. Eating Behaviors, 4, 345-361] was created for the purpose of identifying college students vulnerable to the development of an eating disorder. In a previous study, the COEDS was developed and tested among a college sample based on the continuum model of eating pathology. Following from this initial study, the purpose of the present study was to examine the internal consistency of the final seven-item version as well as its one month test-retest reliability and construct validity. Results indicated that the COEDS performed well when administered in its brief 7-item final format, yielding high internal consistency. Strong test-retest reliability also was observed. Finally, the COEDS demonstrated sound construct validity, showing statistically significant associations with independent measures of disordered eating beliefs and attitudes, as well as with measures assessing disordered eating behaviors. The results of this study provide further support for the use of the COEDS as a measure targeting college students who possess a vulnerability to development of an eating disorder. PMID- 16257813 TI - The comparative utility of statistically derived eating disorder clusters and DSM IV diagnoses: relationship to symptomatology and psychiatric comorbidity at intake and follow-up. AB - INTRODUCTION: The classification of eating disorders has been a matter of considerable debate. The present paper extends previous work and aimed to compare the utility of statistically derived clusters of eating disorders and conventional diagnoses. METHODS: Adult female eating disorder patients who had previously been classified on the basis of cluster analysis of key diagnostic variables were examined on measures of eating disorder symptomatology and psychiatric comorbidity at intake (N=601) and subsequent follow-up after 6 and 36 months (N=349, N=322, respectively). RESULTS: Compared to DSM-IV diagnoses, clusters demonstrated greater utility in terms of more distinct between-group differences and higher effect sizes in relation to a wide range of variables. The greater utility of clusters was in important respects due to the reallocation of EDNOS patients to more relevant alternative categories and to a greater emphasis on psychological and behavioural features of eating disorders. CONCLUSIONS: In order to achieve a better classification of eating disorders, it will be important to place increased emphasis on common psychological features. There is a need to move away from increased use of subtypes and toward a definition of eating disorder per se. PMID- 16257814 TI - Essentials of prescribing in the era of the Medicare Prescription Drug Improvement and Modernization Act. PMID- 16257815 TI - A randomized, placebo-controlled, 6-month study of once-weekly alendronate oral solution for postmenopausal osteoporosis. AB - OBJECTIVE: This study evaluated the overall safety and tolerability of once weekly (OW) alendronate 70 mg oral solution (OS) versus OW placebo OS. METHODS: Postmenopausal, osteoporotic women were enrolled at 51 centers in the United States in a 6-month double-blind, randomized trial. Patients were randomized (1:1) to OW alendronate 70 mg OS or placebo OS. The primary end point was the proportion of patients reporting any upper gastrointestinal (UGI) adverse event (AE) at 6 months. Secondary end points included mean percentage change in urinary N-telopeptide of type I human collagen (NTx) and serum bone-specific alkaline phosphatase (BSAP) at 6 months. RESULTS: Initially, 454 women were enrolled; 392 (86.3%) completed the study. The mean (SD) age was 65.2 (10) years, and the mean (SD) time since menopause was 19.1 (12) years. The proportion of patients experiencing any UGI AE was significantly higher with alendronate OS (23.7%) compared with placebo solution (15.3%), with a treatment difference of 8.3% (95% CI, 0.8%-15.8%; P = 0.024). The proportion of patients experiencing any esophageal AE was 4.0% with alendronate and 3.0% with placebo (treatment difference, 1.0% [95% CI, -2.7% to 4.8%]). In addition, 4.5% of alendronate and 8.7% of placebo patients discontinued the study due to any clinical AE, and 3.3% of alendronate and 1.8% of placebo patients discontinued due to a UGI AE (difference, 1.5% [95% CI, -1.5% to 4.4%]). Alendronate OS produced significantly greater reductions in both NTx and BSAP than placebo (differences, -47.5% and 38.7%, respectively [both, P < 0.001]). CONCLUSIONS: In this 6-month study, patients receiving OW alendronate 70 mg OS had a higher rate of UGI AEs than placebo patients. However, rates of serious UGI AEs, discontinuations due to UGI AEs, and esophageal AEs were similar between groups. UGI AEs in the study were generally mild to moderate in severity and did not result in treatment discontinuation. In addition, OW alendronate 70 mg OS significantly reduced biochemical markers of bone turnover. PMID- 16257816 TI - Effects of rivastigmine treatment on the neuropsychiatric and behavioral disturbances of nursing home residents with moderate to severe probable Alzheimer's disease: a 26-week, multicenter, open-label study. AB - BACKGROUND: Alzheimer's disease (AD) is the most common form of dementia and is characterized clinically by a gradual decline in cognitive performance, an increasingly impaired ability to perform activities of daily living, and neuropsychiatric and behavioral disturbances. OBJECTIVE: The goal of this study was to assess the effect of rivastigmine on the neuropsychiatric and behavioral disturbances of nursing home residents with moderate to severe probable AD and to evaluate the safety and tolerability of rivastigmine in this population. METHODS: This prospective, 26-week, open-label study was conducted in 13 centers in the United States and involved a total of 29 nursing homes. The effects of rivastigmine 3 to 12 mg/d for 26 weeks were assessed in nursing home residents with moderate to severe probable AD. Efficacy was evaluated using the Neuropsychiatric Inventory-Nursing Home (NPI-NH) scale for neuropsychiatric and behavioral disturbances; the Mini-Mental State Examination and the naming subset of the Alzheimer's Disease Assessment Scale-Cognitive subscale for cognitive performance; and the simplified Clinician's Interview-Based Impression of Change Plus Caregiver Input for global functioning. RESULTS: A total of 173 patients (141 women, 32 men; mean [SD]age, 82.6 [5.9] years) were enrolled. After 26 weeks of rivastigmine treatment, the mean (SD) change from baseline for all treated patients in the observed cases population was -2.5 (16.4) (n = 100; P = 0.138); it was -0.8 (16.5) (n = 149; P = 0.576) for the last-observation-carried-forward population. Patients with at least 1 neuropsychiatric symptom present at baseline showed a 3.2-point mean improvement in NPI-NH total score (n = 92; P = 0.062), with 49% of these patients demonstrating a clinically meaningful (ie, > or = 30%) reduction from baseline. At 26 weeks, scores for 8 of the 12 neuropsychiatric and behavioral disturbances in patients with the specific symptom present at baseline showed statistically significant improvements from baseline (delusions [n = 32; P = 0.007], hallucinations [n = 15; P < 0.001], agitation [n = 58; P = 0.044], apathy/indifference [n = 37; P < 0.001], irritability/lability [n = 50; P < 0.001], aberrant motor behavior [n = 32; P < 0.001], nighttime disturbances [n = 22; P < 0.001], and appetite/eating changes [n = 28; P = 0.002]) in the observed cases population. Limitations of this study include that it was open label and not restricted to patients with behavioral disturbances at baseline. CONCLUSION: In the current study, rivastigmine treatment for 26 weeks in nursing home residents with moderate to severe probable AD was associated with decreased NPI NH item scores for a wide range of behavioral disturbances in the subgroup of patients with behavioral symptoms at baseline. PMID- 16257817 TI - Predictors of pneumococcal polysaccharide vaccination among patients at three inner-city neighborhood health centers. AB - BACKGROUND: Despite the availability of pneumococcal polysaccharide vaccine (PPV), vaccination rates are modest, and racial disparity in these rates is known to occur. OBJECTIVE: The purpose of this study was to identify determinants of patient-reported receipt of PPV among patients from 3 inner-city health centers that serve large numbers of minority and disadvantaged patients. METHODS: In 2003, a random sample of patients from 3 inner-city health centers was surveyed by computer-assisted telephone interview concerning their vaccination status and their attitudes and beliefs about PPV. The questionnaire was based on the theory of reasoned action, which emphasizes the contributions to behavioral intention of cognitive beliefs (including the likelihood of consequences), the value of those consequences, and normative beliefs. Factor analysis was conducted to examine whether survey items specific to attitudes about PPV would group together; the variance of the components resulting from the factor analysis was expressed in eigenvalues. Logistic regression analyses were conducted to determine which factors, demographic characteristics, and health habits were independently associated with PPV status. RESULTS: Of 674 patients sent introductory letters (13 of whom were ineligible), 325 (49%) completed the interview with usable data. These patients were primarily aged > or = 65 years (52% [168/325]), female (69% [223/325]), and white (57% [174/306]), and had an annual household income < 20,000 dollars (61% [174/287]). Vaccination status or intention to be vaccinated was unknown for 25 persons, reducing the sample to 300. Of these respondents, 59% (178) reported vaccination, 9% (26) intended to be vaccinated within 6 months, and 32% (96) had no intention of being vaccinated. Vaccination rates were 76% (119/157) for responders aged > or = 65 years and 41% (59/143) for responders aged 50 to 64 years (P < 0.001). Logistic regression analyses found 3 variables that were significantly associated with the likelihood of vaccination: age > or = 65 years; frequency of physician visits; and timing of the last complete physical examination. The factor analysis with varimax rotation identified 2 factors that increased the likelihood of reporting ever receiving PPV: attitudes about PPV and social influences (eigenvalue = 2.69), and beliefs about pneumonia risk and vaccine efficacy (eigenvalue = 1.24). When these factors were added to the model, the effects of age and frequency of seeing a physician became nonsignificant. Compared with those having a complete physical examination < 1 year ago, those having an examination 1 to 2 years ago (odds ratio [OR] = 0.16; 95% CI = 0.04 0.69) or > 2 years ago (OR = 0.10; 95% CI = 0.01-0.78) were less likely to be vaccinated. Those agreeing with the attitudes and social influence factor were more likely to be vaccinated (OR = 8.24; 95% CI = 3.97-17.12), as were those agreeing with the pneumonia risk and vaccine efficacy factor (OR = 2.63; 95% CI = 1.20-5.80). CONCLUSIONS: Attitudes about vaccination and a clinician's recommendation are the most powerful predictors of PPV status. Efforts to raise rates of vaccination and reduce racial disparities should emphasize communication of clinicians' recommendations to patients. PMID- 16257818 TI - Effects of megestrol acetate on pituitary function and end-organ hormone secretion: a post hoc analysis of serum samples from a 12-week study in healthy older men. AB - BACKGROUND: Megestrol acetate (MA) is a synthetic progestin commonly used to promote weight gain in malnourished older individuals. In small studies, MA administration has been associated with reduced serum cortisol concentrations in patients with cancer or AIDS. The impact of MA on the pituitary secretion of adrenocorticotropic hormone (ACTH) and other hormones is unclear, and the prevalence and extent of hypocortisolemia in older individuals after MA treatment is unknown. A randomized, placebo-controlled study of the effects of testosterone (T) and resistance training (RT) on body composition after MA administration in older men has been reported previously. OBJECTIVE: The purpose of this post hoc analysis was to examine the effect of 12 weeks of MA on pituitary function and end-organ hormone secretion in healthy older individuals using frozen serum samples from that study. METHODS: The previous study was conducted at the Department of Geriatrics, Donald W. Reynolds Center on Aging and the General Clinical Research Center at The University of Arkansas for Medical Sciences, Little Rock, Arkansas. Healthy male volunteers aged 60 to 85 years were recruited from the center and were randomly assigned to 1 of 4 study groups: RT + T, T, RT + placebo (P), or P. Subjects enrolled in the RT groups underwent supervised upper- and lower-body strength-training exercises 3 d/wk at 80% of 1 repetition maximum. Subjects in the groups to receive T received injections of testosterone enanthate 100 mg i.m. QW for 12 weeks. Subjects receiving P were given 1-mL saline injections i.m. QW for 12 weeks. All subjects received MA 800 mg p.o. QD concurrently for 12 weeks. For the present analysis, serum concentrations of the pituitary hormones follicle-stimulating hormone (FSH), thyroid-stimulating hormone (TSH), ACTH, prolactin (PRL), and luteinizing hormone (LH), as well as the end-organ hormones estradiol (E2), cortisol, free T4, and T, were measured in samples obtained at baseline (week 0) and after 12 weeks of MA treatment. RESULTS: Serum samples from 21 men (mean [SD]age, 67.0 [7.3]years; mean [SD] body mass index, 23.1 [10.4] kg/m2; mean [SD] percentage of body fat, 22.5% [8.8%]; RT + T, T, RT + P, and P groups, n = 4, 5, 6, and 6 subjects, respectively) were available from the original study. The mean percentage changes from baseline in serum pituitary hormone concentrations after 12 weeks of MA administration were as follows: TSH, -14.7%; ACTH, -89.5%; PRL, 162.2%; and LH, -49.0%; (P = 0.03, <0.001, <0.001, and <0.001, respectively). The mean (SD) percentage changes from baseline in serum end-organ hormone concentrations with MA at 12 weeks were as follows: E2, 181.6%; and cortisol, -90.8% (both, P < 0.001). In the P and RT + P groups, the mean percentage changes from baseline in T were -84% and -85%, respectively (both, P < 0.001). FSH and free T4 concentrations were not significantly changed. CONCLUSIONS: This analysis of serum samples from healthy older men suggests that MA administration significantly affected the secretion of several pituitary hormones and end-organ hormone synthesis. Most notably, ACTH secretion and serum cortisol levels were statistically significantly suppressed in 20 of 21 subjects, without the development of clinically significant adrenal suppression. PMID- 16257819 TI - Factors associated with potentially inappropriate drug utilization in a sample of rural community-dwelling older adults. AB - BACKGROUND: Many medications present special risks when used by older adults (ie, those aged > or = 65 years) and are considered potentially inappropriate for this population. The Beers criteria are often used to identify such medications. Past research has documented that use of Beers drugs is common among older adults. OBJECTIVE: The aim of this work was to examine factors associated with potentially inappropriate drug use among rural community-dwelling older adults using a conceptual framework adapted from the Andersen-Newman behavioral model of health service use. METHODS: This was a population-based, cross-sectional survey. Data were collected via face-to-face home interviews between 2002 and 2004. Rural community-dwelling older adults residing in a single county in North Carolina were eligible. Potentially inappropriate drug use was operationalized using the Beers criteria. Data concerning predisposing (ie, age, sex, race, education, and marital status), enabling (ie, social support and insurance status), need (ie, disability and history of major depression, hypertension, osteoarthritis, back problems, or other comorbidities), and utilization factors (ie, number of medications used) were collected. RESULTS: Data were gathered from 892 people, with information on medication use available for 800. Two hundred thirteen of these 800 participants (26.6%) used > or = 1 Beers drug. Compared with individuals who used no Beers drugs, those who used > or = 1 Beers drug reported lower levels of social support (odds ratio [OR], 0.94; 95% CI, 0.90-0.99) and higher levels of disability (OR, 1.48; 95% CI, 1.11-1.97), used more medications (OR, 1.07; 95% CI, 1.01-1.13), and were more likely to have a history of major depression (OR, 1.67; 95% CI, 1.05-2.66), hypertension (OR, 1.58; 95% CI, 1.07 2.33), osteoarthritis (OR, 1.58; 95% CI, 1.09-2.29), and back problems (OR, 1.72; 95% CI, 1.19-2.47). CONCLUSION: As suggested by the Andersen-Newman model, the risk of potentially inappropriate drug use is highest among those with the greatest medication needs, as evidenced by poorer health status in this sample of rural community-dwelling older patients. PMID- 16257820 TI - Antiepileptic drug use and the occurrence of pressure ulcers among bedridden institutionalized elderly patients: a retrospective chart review. AB - BACKGROUND: Phenytoin (PH) is indicated primarily for the control of grand mal and psychomotor seizures. However, topical PH has been used for the treatment of various types of ulcers, including pressure ulcers. OBJECTIVE: The aim of this study was to investigate the possibility of a relationship between the use of oral PH and the prevalence of pressure ulcers among bedridden institutionalized elderly patients. METHODS: This retrospective chart review was conducted in a state-run urban geriatric medical center in Israel and involved long-term bedridden institutionalized patients who were receiving chronic antiepileptic medication during the 7-year period between January 1996 and December 2003. The prevalence of pressure ulcers in patients who received treatment with PH alone or in combination with other antiepileptic drugs was compared with that in patients who received antiepileptic agents other than PH. RESULTS: The study analyzed data from the medical charts of 153 patients, 72 of whom received PH alone or in combination with other antiepileptic drugs, and 81 of whom received antiepileptic agents other than PH. Patients' mean (SD) age was 78.5 (7.2) years; 106 (69.3%) were women. All patients were totally dependent with respect to activities of daily living (mean Katz score, 2.0 [2.0]) and had severe cognitive decline (mean Mini-Mental State Examination score, 3.5 [3.3]). Pressure ulcers occurred in 9.7% of PH recipients and 27.2% of non-PH recipients (P = 0.006; chi2 = 7.55). In PH recipients, 85.7% of pressure ulcers were of mild to moderate severity (stage I or II), compared with 59.1% of ulcers in non-PH recipients; the difference between groups was not statistically significant. Four (18.2%) non-PH recipients and no PH recipients had stage IV pressure ulcers. In the PH group, 71.4% of patients had a pressure ulcer in only 1 anatomic location, compared with 22.7% of the non-PH group (P = 0.023; chi2 = 5.13); 28.6% of PH recipients and 63.6% of non-PH recipients had pressure ulcers in 2 or 3 anatomic locations; and 3 (13.6%) non-PH recipients and no PH recipients had pressure ulcers in > or = 4 areas. CONCLUSIONS: In the long-term bedridden institutionalized patients studied, those who received PH had lower rates of pressure ulcers, as well as less severe ulcers. PH may be a useful anticonvulsive agent in frail elderly patients, who are at risk for the development of pressure ulcers. PMID- 16257821 TI - National trends in and predictors of propoxyphene use in community-dwelling older adults. AB - BACKGROUND: Several pain management guidelines and explicit medication-use criteria identify propoxyphene as an inappropriate medication for use in older adults. OBJECTIVE: This study was conducted to estimate trends in propoxyphene use among community-dwelling elderly (age > or = 65 years) Medicare beneficiaries from 1993 through 1999 and to determine whether beneficiaries' drug coverage and specific characteristics of their physicians were associated with receipt of propoxyphene in 1999. METHODS: Data from the Medicare Current Beneficiary Survey (MCBS) were used to examine the prevalence of propoxyphene use in cross-sections of nationally representative samples of community-dwelling elderly Medicare beneficiaries from 1993 through 1999. The 1999 MCBS was linked with the 1999 Area Resource File to examine patient and physician factors associated with propoxyphene use in the community-dwelling elderly at the county level. RESULTS: Rates of propoxyphene use were generally stable over the 7-year period, from an annual prevalence of 6.8% in 1993 to the slightly decreased prevalence of 6.6% in 1999. No protective effects against propoxyphene use were observed based on beneficiaries' drug coverage or type of drug coverage. Rather, Medicaid beneficiaries were more likely to receive propoxyphene than those without drug coverage (odds ratio [OR] = 1.40; 95% CI, 1.02-1.92). Among physician characteristics, male sex (OR = 1.34; 95% CI, 1.02-1.75) and medical specialty (OR = 0.81; 95% CI, 0.65-1.00) were strongly correlated with prescribing of propoxyphene. CONCLUSION: This study found a continuing high prevalence of propoxyphene use in the community-dwelling elderly Medicare population from 1993 through 1999, with > 2 million beneficiaries receiving the drug in 1999. PMID- 16257822 TI - Self-reported medication use in community-residing older adults: A pilot study. AB - BACKGROUND: Older adults (ie, those aged > or = 65 years) are at increased risk of developing drug therapy problems, which may lead to poor health outcomes and decreased quality of life. OBJECTIVE: The primary goal of this pilot study was to evaluate and report medication use and potential drug therapy problems in older adults who received Eldercare program assistance through the Orange County Department on Aging in North Carolina. METHODS: Between May and July 2002, subjects were consecutively sampled from a registry of adults aged > or = 60 years enrolled in the Eldercare program. To be eligible for the study, individuals had to be receiving the services of the Eldercare program, speak English, and reside independently in the community of Orange County. The older adults were contacted by the program director to determine interest in participating in the study. If interested, the individual was contacted by a trained pharmacy doctoral student to verify study eligibility and arrange a home visit. At the home visit, information was collected via self-report on medication use and medical history. Nonadherence, potentially inappropriate prescribing, health literacy, and functional capacity were also assessed. All home visits were conducted between May and July 2002. RESULTS: A total of 100 subjects were interviewed. The mean (SD) age of respondents was 77.5 (8.7) years; 85% were women, 66% were white, 34% black, and 70% lived alone. The mean (SD) number of prescription medications used per patient was 9.6 (4.1). Adequate health literacy, defined as a score > or = 23 (range of possible scores, 0-36) on the Short Test of Functional Health Literacy in Adults, was documented in only 35% of individuals. Twenty-five percent of the sample (25/100) had reduced functional capacity when evaluated on the Functional Activities Questionnaire, with total scores > or = 10 (range of possible scores, 10-30) indicative of reduced functional ability. Rates of nonadherence, defined as a score of 0 to 3 on the 4 item Morisky instrument, were 53%. When evaluating inappropriate prescribing based on the Beers criteria, 34% of individuals used > or = 1 potentially inappropriate medication. In bivariate analyses, there was a statistically significant relationship between race and number of medications (P < 0.002), adherence (P < 0.001), health literacy status (P < 0.001), and functional capacity (P < 0.027). No differences were noted when examining the effects of age on the same variables. CONCLUSIONS: In this study, older adults residing independently in the community were responsible for managing and taking a considerable number of medications on a daily basis. In addition, inadequate health literacy, poor adherence, and potentially inappropriate medication use were prevalent in this sample. The results also suggest that race may have played an important role in the risk of developing medication-related problems. PMID- 16257825 TI - Assessing fetal cardiac ventricular function. AB - Fetal echocardiography has been used primarily to identify fetuses with structural malformations of the heart. Evaluation of fetal ventricular function, however, has received minimal attention since the inception of fetal echocardiography in the early 1980s. This communication reviews the use of M mode, B-mode and pulsed Doppler ultrasound to examine cardiac function. M-mode ultrasound is used to determine the size of the fetal heart, the end-diastolic and end-systolic dimensions of the ventricular chambers, and the thickness of the ventricular walls and the interventricular septum, and to measure the diameter of the mitral and tricuspid valves as well as the diameter of the aorta and pulmonary artery. B-mode evaluation of the fetal heart includes measurement of atrial and ventricular dimensions as well as dimensions of the outflow tracts. This modality is useful when M-mode measurements cannot be made due to fetal position. Once measurements of cardiac structures are obtained using either M mode or B-mode ultrasound, pulsed Doppler recording of mitral valve, tricuspid valve, aortic valve and pulmonary artery waveforms can be used to compute cardiac output as well as stroke volume. In addition, pulsed Doppler can be used to evaluate diastolic and systolic cardiac functions by examining the components of each waveform. PMID- 16257826 TI - Ribosomal proteins in the spotlight. AB - The assignment of specific ribosomal functions to individual ribosomal proteins is difficult due to the enormous cooperativity of the ribosome; however, important roles for distinct ribosomal proteins are becoming evident. Although rRNA has a major role in certain aspects of ribosomal function, such as decoding and peptidyl-transferase activity, ribosomal proteins are nevertheless essential for the assembly and optimal functioning of the ribosome. This is particularly true in the context of interactions at the entrance pore for mRNA, for the translation-factor binding site and at the tunnel exit, where both chaperones and complexes associated with protein transport through membranes bind. PMID- 16257827 TI - Endoplasmic reticulum stress and the making of a professional secretory cell. AB - Homeostasis of the protein folding machinery in the endoplasmic reticulum (ER) is maintained via several parallel unfolded protein response pathways that are remarkably conserved from yeast to man. Together, these pathways are integrated into a complex circuitry that can be modulated in various ways, not only to cope with various stress conditions, but also to fine-tune the capacity of the ER folding machinery when precursor cells differentiate into professional secretory cells. PMID- 16257828 TI - The ribosomal peptidyl transferase center: structure, function, evolution, inhibition. AB - The ribosomal peptidyl transferase center (PTC) resides in the large ribosomal subunit and catalyzes the two principal chemical reactions of protein synthesis: peptide bond formation and peptide release. The catalytic mechanisms employed and their inhibition by antibiotics have been in the focus of molecular and structural biologists for decades. With the elucidation of atomic structures of the large ribosomal subunit at the dawn of the new millennium, these questions gained a new level of molecular significance. The crystallographic structures compellingly confirmed that peptidyl transferase is an RNA enzyme. This places the ribosome on the list of naturally occurring ribozymes that outlived the transition from the pre-biotic RNA World to contemporary biology. Biochemical, genetic and structural evidence highlight the role of the ribosome as an entropic catalyst that accelerates peptide bond formation primarily by substrate positioning. At the same time, peptide release should more strongly depend on chemical catalysis likely involving an rRNA group of the PTC. The PTC is characterized by the most pronounced accumulation of universally conserved rRNA nucleotides in the entire ribosome. Thus, it came as a surprise that recent findings revealed an unexpected high level of variation in the mode of antibiotic binding to the PTC of ribosomes from different organisms. PMID- 16257829 TI - The endoplasmic reticulum in xenobiotic toxicity. AB - The endoplasmic reticulum (ER) is involved in an array of cellular functions that play important roles in xenobiotic toxicity. The ER contains the majority of cytochrome P450 enzymes involved in xenobiotic metabolism, as well as a number of conjugating enzymes. In addition to its role in drug bioactivation and detoxification, the ER can be a target for damage by reactive intermediates leading to cell death or immune-mediated toxicity. The ER contains a set of luminal proteins referred to as ER stress proteins (including GRP78, GRP94, protein disulfide isomerase, and calreticulin). These proteins help regulate protein processing and folding of membrane and secretory proteins in the ER, calcium homeostasis, and ER-associated apoptotic pathways. They are induced in response to ER stress. This review discusses the importance of the ER in molecular events leading to cell death following xenobiotic exposure. Data showing that the ER is important in both renal and hepatic toxicity will be discussed. PMID- 16257830 TI - Function and regulation of multidrug resistance proteins (MRPs) in the renal elimination of organic anions. AB - The reabsorptive and excretory capacity of the kidney has an important influence on the systemic concentration of drugs. Multidrug resistance proteins (MRP/ABCC) expressed in the kidney play a critical role in the tubular efflux of a wide variety of drugs and toxicants, and, in particular, of their negatively charged phase II metabolites. Nine structurally and functionally related MRP family members have been identified (MRP1-9), which differ from each other by their localization, expression levels, and substrate specificity. During altered physiological circumstances, adaptations in these transporters are required to avoid systemic toxicity as well as renal tubular damage. Key players in these events are hormones, protein kinases, nuclear receptors, and disease conditions, which all may affect transporter protein expression levels. This review discusses current knowledge on the renal characteristics of MRP1-9, with specific focus on their regulation. PMID- 16257832 TI - New horizon of MDR1 (P-glycoprotein) study. AB - MDR1 (once P-glycoprotein, now referred to as ABCB1) plays a role as a blood brain barrier, preventing drug absorption into the brain, and is known to confer multiple drug resistance in cancer chemotherapy. MDR1 is composed of two repeated fragments, and there are six transmembrane domains (TMD) on the N-terminal of each repeat and a nucleotide (ATP) binding domain (NBD) on the C-terminal. These two repeats are dependent but cooperate as one functional molecule, with one pocket for excreting drugs. The 12 TM domains form a funnel facing the outside of cells, and NBD is in cytosol as a dimer. One NBD is composed of the Walker A, Q loop, ABC-signature and the Walker B for phosphate binding of nucleotide. This tertiary structure of MDR1 is suggested from the structure of the NBD of histidine permease (HisP), clarified by x-ray crystallography. On the model of HisP, the NBD positions described above make a functional domain, and the same NBD structure is found on many other ABC transporters. An experiment with MDR1 gene knockout mice showed the high plasma AUC of drugs in mdr null mice [mdr1a(-/ )] and a high level in the brain, indicating that MDR1 has an efflux function (prevention of absorption) in the intestinal lumen and acts as a barrier of drug uptake in the brain, as well as has the function of urinary and biliary excretion of drugs. The transcription of MDR1 is dependent on two sites; the promoter site (-105/-100)(-245/-141) and the enhancer site (-7864/-7817). Autoantibody from autoimmune hepatitis patients weakly reacted with the extracellular peptide (aa314-aa328 between TM5 and 6) of MDR1 on the outside of the cell membrane, and did not react with peptides in the NBD and in the membrane-spanning region in TM5. There is an ambiguity about the function of MDR1 as GlcCer translocase. PMID- 16257831 TI - Studying cellular processes and detecting disease with protein microarrays. AB - Protein microarrays are a rapidly developing analytic tool with diverse applications in biomedical research. These applications include profiling of disease markers or autoimmune responses, understanding molecular pathways, protein modifications, and protein activities. One factor that is driving this expanding usage is the wide variety of experimental formats that protein microarrays can take. In this review, we provide a short, conceptual overview of the different approaches for protein microarray. We then examine some of the most significant applications of these microarrays to date, with an emphasis on how global protein analyses can be used to facilitate biomedical research. PMID- 16257833 TI - Arylamine N-acetyltransferases: what we learn from genes and genomes. AB - Arylamine N-acetyltransferases (NATs) are phase II xenobiotic metabolizing enzymes, catalyzing acetyl-CoA-dependent N- and O-acetylation reactions. All NATs have a conserved cysteine protease-like Cys-His-Asp catalytic triad inside their active site cleft. Other residues determine substrate specificity, while the C terminus may control hydrolysis of acetyl-CoA during acetyltransfer. Prokaryotic NAT-like coding sequences are found in >30 bacterial genomes, including representatives of Actinobacteria, Firmicutes and Proteobacteria. Of special interest are the nat genes of TB-causing Mycobacteria, since their protein products inactivate the anti-tubercular drug isoniazid. Targeted inactivation of mycobacterial nat leads to impaired mycolic acid synthesis, cell wall damage and growth retardation. In eukaryotes, genes for NAT are found in the genomes of certain fungi and all examined vertebrates, with the exception of canids. Humans have two NAT isoenzymes, encoded by highly polymorphic genes on chromosome 8p22. Syntenic regions in rodent genomes harbour two Nat loci, which are functionally equivalent to the human NAT genes, as well as an adjacent third locus with no known function. Vertebrate genes for NAT invariably have a complex structure, with one or more non-coding exons located upstream of a single, intronless coding region. Ubiquitously expressed transcripts of human NAT1 and its orthologue, murine Nat2, are initiated from promoters with conserved Sp1 elements. However, in humans, additional tissue-specific NAT transcripts may be expressed from alternative promoters and subjected to differential splicing. Laboratory animals have been widely used as models to study the effects of NAT polymorphism. Recently generated knockout mice have normal phenotypes, suggesting no crucial endogenous role for NAT. However, these strains will be useful for understanding the involvement of NAT in carcinogenesis, an area extensively investigated by epidemiologists, often with ambiguous results. PMID- 16257834 TI - Clinical pharmacogenetics of irinotecan (CPT-11). AB - Irinotecan (CPT-11) is now widely used, especially for colorectal and lung cancers, whereas the drug causes severe adverse drug reactions (ADR), such as leukopenia/neutropenia or diarrhea. Irinotecan undergoes drug metabolism to form an active SN-38, which is further converted to its beta-glucuronide by UDP glucuronosyltransferase (UGT) 1A1. A variant in the promoter of UGT1A1 gene, UGT1A1*28 allele, has been extensively studied, and pharmacogenetic relationships between the variant and ADR to irinotecan have been reported. A case-control study of Japanese cancer patients demonstrated that the patients having UGT1A1*28 were at significantly increased risk of severe ADR to irinotecan. To date, genetic variations of the UGT1A1 gene is the most important hereditary factor to predict severe ADR to irinotecan. The UGT1A1*28 is the only one variant that has multiple lines of clinical evidence in multiple races, whereas genetic variations of other UGT isoforms, drug-metabolizing enzymes and drug transporters need more confirmations of its clinical significance in multiple patient groups. At present, irinotecan chemotherapy based on a patient's UGT1A1 genetic status is scientifically reasonable. PMID- 16257835 TI - Enuresis alarm treatment. AB - OBJECTIVE: Treatment for childhood nocturnal enuresis emphasizes either a psychological or pharmacological approach. The enuresis alarm, in comparative studies, has emerged as the most effective psychological treatment. In this review we investigate both outcome rates and influential factors from recently published studies. MATERIAL AND METHODS: A search of papers published between 1980 and 2002 in the English language involving at least 10 children in which the enuresis alarm was employed as a stand-alone intervention revealed 38 studies. RESULTS: Heterogeneity in terms of inclusion and outcome parameters made comparison between studies problematic. The most frequently adopted definitions were "wet at least 3 times a week" in terms of severity at inclusion, "14 consecutive dry nights" as a success criterion and "> 1 wet night a week" as a relapse criterion. Success rates across all studies ranged from 30% to 87% and were influenced by the type of enuresis, the treatment duration and the success criteria adopted. In an homogenous subset of 20 studies, 65% success with alarm treatment was found. Further analysis revealed equivalence between different forms of alarm, pre- and within-treatment predictors of outcome and possible mode of action. Relapse rates (ranging between 4% and 55%) were reported in 20 studies, with an homogeneous subset indicating that 42% of children relapsed following alarm treatment. CONCLUSIONS: The enuresis alarm is an effective intervention for children with nocturnal enuresis. There are a number of factors, both pre- and within-treatment, that appear to influence its effectiveness and may assist clinical decisions concerning its appropriateness for any particular child. PMID- 16257836 TI - Effects of mepartricin, a polyene macrolide antibiotic, on estrogen-induced hyperplastic growth of the dorsolateral prostate and seminal vesicle in immature castrated rats. AB - OBJECTIVE: To investigate the effects of mepartricin, a polyene macrolide antibiotic, on estrogen-induced hyperplastic prostate and seminal vesicle (SV) growth in castrated rats. MATERIAL AND METHODS: Immature rats aged 3 weeks were castrated and left untreated for 1 week. Then, 17beta-estradiol benzoate (E(2) BA) was subcutaneously injected at a dose of 10 microg/day twice weekly, and mepartricin was orally administered at doses of 2.5, 5 and 10 mg/kg once daily for 3 weeks. The weights and hydroxyproline contents of the prostate and SV, the activity of growth factors (GFs) in the dorsolateral prostate (DLP) and the serum estrogen level were measured. Histological examination of the prostate and SV was also performed. RESULTS: Mepartricin dose-dependently suppressed the increase in the serum estrogen level, the weights and hydroxyproline contents of the DLP and SV and the elevation of GF activity in the DLP induced by E(2)-BA treatment. Histological examination also revealed that treatment with mepartricin reduced collagen accumulation and thickening of the smooth muscle layer in the DLP and SV, and proliferation of the glandular epithelium in the DLP. CONCLUSIONS: These results suggest that mepartricin suppresses hyperplastic growth of the DLP and SV induced by estrogen in immature castrated rats, the underlying mechanism being a reduction in the serum estrogen level, thereby suppressing stromal cell proliferation and activation. PMID- 16257838 TI - Geographical variation in incidence of prostate cancer in Sweden. AB - OBJECTIVE: To investigate the geographical variation in prostate cancer incidence in Sweden, in particular the incidences of screening-detected tumours and curative treatment of prostate cancer. MATERIAL AND METHODS: Data were retrieved from the National Prostate Cancer Register of Sweden for all cases of prostate cancer diagnosed in the year 2000-01. There were a total of 14 376 cases of prostate cancer and the mean total annual age-adjusted incidence was 197/100 000 men. There were 3318 cases in tumour category T1c, i.e. non-palpable tumours diagnosed during work-up for an elevated serum level of prostate-specific antigen, 1006 of which (30%) were asymptomatic and detected at a health check-up. RESULTS: The difference between the counties with the lowest and highest age adjusted incidences per 100 000 men of total prostate cancer was almost twofold (128 vs 217). The corresponding variation in incidence of category T1c tumours was more than fourfold (13 vs 60); the difference in incidence of T1c tumours detected in asymptomatic men was up to 10-fold (2 vs 20); and there was more than a fourfold variation in incidence of curative treatment between counties (13 vs 67). Measured incidences were mostly highest in urban regions and in counties with university hospitals. CONCLUSION: There are large geographical variations in prostate cancer incidence and in the frequency of curative treatment for prostate cancer in Sweden and there appear to be large geographical variations in the uptake of prostate cancer screening. PMID- 16257837 TI - Transforming growth factor-beta1 regulates cell growth and causes downregulation of SMemb/non-muscle myosin heavy chain B mRNA in human prostate stromal cells. AB - OBJECTIVES: SMemb/non-muscle myosin heavy chain B (SMemb/NMMHC-B) is most abundantly expressed in proliferating smooth muscle cells and correlates with phenotypic changes from a contractive to a proliferative type. The stromal cells of the prostate play a crucial role in the regulation of prostatic growth and function. The aim of this study was to investigate the effects of the multifunctional cytokine transforming growth factor-beta1 (TGF-beta1) on SMemb/NMMHC-B mRNA expression and stromal cell growth. The expression of the SM2 isoform of smooth muscle myosin heavy chain (SMMHC) mRNA was also examined. MATERIAL AND METHODS: Primary cultures of prostate stromal cells were established by means of an explant method from eight normal prostates. The effects of TGF beta1 on stromal cell growth were determined by means of a 3-(4,5-dimethylthiazol 2-yl)-2,5-diphenyl tetrazolium bromide conversion assay. SMemb/NMMHC-B and SM2 mRNA expression were analyzed quantitatively by means of real-time polymerase chain reaction. RESULTS: In the absence of TGF-beta1, cells expressed alpha smooth muscle actin and vimentin. After TGF-beta1 treatment, the expression of alpha-smooth muscle actin increased and cells also expressed desmin. TGF-beta1 at concentrations of 1.0, 5.0 and 10 ng/ml suppressed cell growth by 72%, 62% and 56%, respectively, downregulated SMemb/NMMHC-B mRNA expression by 71%, 52% and 38%, respectively and upregulated SM2 mRNA expression 2.1-, 3.0- and 5.3-fold, respectively. CONCLUSIONS: These results demonstrate that TGF-beta1 modulates the smooth muscle cell phenotype from a proliferative to a contractile type and that the inhibitory effects of TGF-beta1 on stromal cell growth correlate with downregulation of the SMemb/NMMHC-B gene. PMID- 16257839 TI - Prediction of the volume of large prostate cancers by multiple core biopsies. AB - OBJECTIVE: To evaluate whether large-volume prostate cancers can be predicted by means of multiple needle biopsies. MATERIAL AND METHODS: In 115 men, 8-14 (mean 10) biopsies were taken, including eight from standardized positions (apex, mid medial, mid-lateral and base). Biopsies were reviewed, the length of the cancer measured and the percentage cancer length calculated. All men underwent radical prostatectomy. The prostatectomy specimens were totally embedded and the tumor volume was measured planimetrically. The predictive values of the number and percentage of cores positive for cancer, cancer length and percentage cancer length were calculated for tumor volumes of >4, >6 and >8 ml. RESULTS: Using univariate logistic regression, cancer length and percentage cancer length predicted tumor volumes of >4 (p<0.001), >6 (p<0.001) and >8 ml (p<0.05). These measures were better predictors of tumor volume than the number and percentage of cores positive for cancer. A biopsy cancer length of > or =30 mm and a percentage cancer length of > or =25% predicted a tumor volume of >4 ml in 95% and 93% of cases, respectively. For tumor volumes of >6 or >8 ml, predictive values were lower. Tumor volumes of <2 and <4 ml were found in 13% and 35%, respectively of men with as many as six positive cores, indicating that the number of positive cores was less useful as a predictor of tumor volume than the cancer length. CONCLUSIONS: Cancer length and percentage cancer length are significant predictors of large tumor volumes. It is recommended that the linear extent of cancer in prostate biopsies should be reported by the pathologist. PMID- 16257841 TI - Changes in tissue prostatic acidic phosphatase during endocrine treatment of patients with prostatic carcinoma. AB - OBJECTIVE: We have previously developed methods for the quantification of different macromolecules in aspiration biopsy material and described the changes in prostate-specific antigen (T-PSA) during cancer treatment. We have now studied the changes in tissue prostatic acidic phosphatase (T-PAP) in 58 endocrine treated patients with prostatic carcinoma and compared these data with cancer development data and tissue PSA (T-PSA) levels. MATERIAL AND METHODS: PAP and PSA were quantified in aspiration biopsies taken before treatment and after 6 and 12 months of treatment. Patients were followed until death or for >98 months. RESULTS: Pretreatment T-PSA was more strongly associated with survival than T PAP. Both T-PSA and T-PAP decreased in responders during treatment. In non responders, T-PSA and T-PAP increased after 12 months in 17/18 and 7/13 patients, respectively. Estrogen-treated responders had significantly higher T-PSA, but not T-PAP, treatment values than those treated with orchidectomy or gonadotropin releasing hormone. CONCLUSIONS: The inferiority of serum PAP compared to PSA for monitoring cancer treatment may reflect its less pronounced changes at the tissue level, indicating different in vivo regulation of the two markers. Estrogen stimulation of PSA synthesis in vivo may underlie the higher PSA levels observed during estrogen treatment. PMID- 16257840 TI - The significance of tumor heterogeneity for prediction of DNA ploidy of prostate cancer. AB - OBJECTIVE: In a previous study, we mapped the ploidy heterogeneity of prostate cancer using flow cytometry in 676 tumor samples from 50 radical prostatectomy specimens. Ploidy heterogeneity was common (42% of tumors) and was found in all non-diploid tumors. The volume of non-diploid tumor was estimated and found to predict extra-prostatic extension and seminal vesicle invasion. The aim of this study was to evaluate the impact of tumor heterogeneity on preoperative ploidy assessment. MATERIAL AND METHODS: In 50 men at least six core biopsies were taken before prostatectomy. Sections from biopsies with cancer were Feulgen-stained for image cytometry. After exclusion of biopsies with insufficient material, 123 histograms from 48 men (mean 2.6; range 1-7) remained for analysis. RESULTS: In 32 men, biopsies were diploid. In 16 men, at least one biopsy was non-diploid (14 tetraploid, two aneuploid) and 10 of them also had diploid biopsies. In 34 men (71%), the prostatectomy specimens were correctly predicted as being either diploid (48%) or non-diploid (23%). The sensitivity and specificity of biopsies for predicting non-diploid cancer were 55% and 82%, respectively, and the positive and negative predictive values were 69% and 72%, respectively. The ploidy status of tumors with and without ploidy heterogeneity was correctly predicted in 55% and 82% of cases, respectively (p=0.04). Biopsies underestimated ploidy in 9/20 tumors (45%) with heterogeneous ploidy status. Underestimation mainly occurred when one or two cores were analyzed. CONCLUSIONS: Preoperative prediction of the ploidy status of prostate cancer is hampered by tumor heterogeneity. Analysis of multiple biopsies is important for correct preoperative ploidy estimation. PMID- 16257842 TI - Hemorheological and hemodynamic changes in predialysis patients after normalization of hemoglobin with epoetin-alpha. AB - OBJECTIVE: Changes in blood viscosity and total peripheral resistance may contribute to increased blood pressure during partial correction of renal anemia with erythropoietin. An increase in hemoglobin level is followed by decreases in cardiac output and left ventricular mass. We examined how normalization of hemoglobin in predialysis patients affects both hemorheological and hemodynamic variables. MATERIAL AND METHODS: Twelve moderately anemic predialysis patients (hemoglobin 115.9+/-7.8 g/l) received epoetin-alpha with the aim of achieving a normal hemoglobin level (135-160 g/l). Hemorheological variables were measured using rotational viscometry. Cardiac index was determined by means of Doppler echocardiography. RESULTS: After 48 weeks, the hematocrit level had increased from 37.9%+/-3.0% to 47.0%+/-3.1% (p<0.0001). Blood viscosity increased from 3.84+/-0.33 to 4.59+/-0.4 mPa x s (p<0.001). Blood viscosity standardized to a hematocrit level of 45% and a plasma viscosity of 1.31 mPa x s did not change. Plasma viscosity, erythrocyte aggregation tendency and erythrocyte fluidity remained unchanged. The cardiac index decreased from 2.64+/-0.57 to 2.19+/-0.72 l/min/m(2) (p<0.05). The total peripheral resistance index increased from 3270+/ 985 to 4013+/-1046 (dyn x s/cm(5))m(2) (p<0.05). Blood pressure remained constant, but the amount of antihypertensive medication used increased by 30%. CONCLUSIONS: Hemoglobin normalization in predialysis patients raised blood viscosity and total peripheral resistance due to an increase in hematocrit level, without other consistent hemorheological changes. Antihypertensive therapy had to be increased in many patients to maintain an acceptable blood pressure. The cardiac index was reduced, which may have prevented further development of left ventricular hypertrophy. PMID- 16257843 TI - Is there any relationship between serum levels of interleukin-10 and atherosclerosis in hemodialysis patients? AB - OBJECTIVE: Cardiovascular complications due to atherosclerosis (AS) are the major cause of mortality in hemodialysis (HD) patients. Inflammation may play an important role in the development of AS. Several studies have demonstrated an association between AS and acute-phase proteins and cytokines in the general population and in HD patients. Interleukin-10 (IL-10) is an anti-inflammatory cytokine. The aim of this study was to compare serum levels of inflammatory and anti-inflammatory indicators in HD patients according to the presence or absence of AS. MATERIAL AND METHODS: A total of 33 HD patients were studied. AS was defined as the presence of plaques as detected by Doppler ultrasonography. The patients were subgrouped according to the presence or absence of plaques. Serum levels of IL-1, -2, -6 and -10, C-reactive protein (CRP) and tumor necrosis factor-alpha (TNF-alpha) were measured. Risk factors for AS, such as age, gender, hypertension, hyperlipidemia and duration of HD, were also evaluated. RESULTS: Patients with AS had significantly higher high sensitivity (hs)-CRP and lower IL 10 levels. Blood pressure was also elevated in patients with AS. There was an inverse correlation between CRP and IL-10 levels in patients with AS. CONCLUSION: Patients with AS undergoing HD had low serum levels of the anti-inflammatory cytokine IL-10 and high serum levels of hs-CRP. These results may suggest that limitation of the anti-inflammatory response in atherosclerotic uremic patients is a triggering or contributory factor for AS. PMID- 16257844 TI - Do interleukin-6, hyaluronan, soluble intercellular adhesion molecule-1 and cancer antigen 125 in dialysate predict changes in peritoneal function? A 1-year follow-up study. AB - OBJECTIVE: Diminishing ultrafiltration and dialysis adequacy may limit the long term use of peritoneal dialysis (PD). Inflammation may play a role in changes in peritoneal function. This study was designed to evaluate alterations in peritoneal function and soluble factors in dialysate during a 1-year follow-up period. MATERIAL AND METHODS: A personal dialysis capacity test was performed at the start of the study and after 6 and 12 months in 20 patients in order to determine dialysis adequacy and membrane characteristics. Dialysate was collected during the test days for analyses of interleukin-6 (IL-6), soluble intercellular adhesion molecule-1, hyaluronan and cancer antigen 125 (CA125). RESULTS: There were no significant changes in dialysis adequacy or membrane characteristics during the 1-year follow-up period. The appearance rate of IL-6 in dialysate increased significantly (419.8+/-63.3 at the start, 784.1+/-136.4 after 6 months and 1149.3+/-252.2 ng/24 h after 12 months; p=0.006) during follow-up. Furthermore, the appearance rate of CA125 increased throughout the study in patients using icodextrin, but decreased slightly in patients using only conventional dialysis solutions. CONCLUSIONS: There were no major changes in dialysis adequacy or membrane characteristics during the follow-up period, but increased IL-6 in dialysate may reflect peritoneal inflammation, which may lead to long-term alterations in the peritoneal membrane. Icodextrin may have a preventive effect on the longevity of the peritoneal membrane. PMID- 16257845 TI - Discontinuation of dialysis treatment: experience of a single dialysis centre. AB - OBJECTIVE: Discontinuation of dialysis is a common cause of death in end-stage renal disease (ESRD) patients in the USA and UK, but is less common in the rest of Europe and in Japan. The aim of this study was to describe the discontinuation pattern in a single dialysis unit in eastern Finland. MATERIAL AND METHODS: We retrospectively analysed the case history and cause of death of 146 dialysis patients in whom dialysis treatment was started between 1992 and 2001 and who had died by March 2003. We compared patients who died after withdrawal from dialysis and those who continued dialysis until death. RESULTS: In 53 patients (36.3%) dialysis treatment was discontinued before death (withdrawal group). In the rest of the patients (control group; n=93) dialysis was continued until death. The patients in the withdrawal group were older (median 69 vs 65 years at the onset of ESRD), more often institutionalized before death (49% vs 11.8%) and more often had dementia diagnosed before death (20.8% vs 2.2%) than those in the control group. They were also less rehabilitated before death (54.7% vs 76.7%) and their treatment more often lasted for <3 months (20.8% vs 7.6%). The patients in the withdrawal group died less often of cardiac disease (11.3% vs 39.8%), whereas kidney disease was the commonest cause of death (41.5 vs 19.4%). The commonest reason for discontinuation of dialysis was severe medical illness (86.5%). In most cases the nephrologist or the renal team raised the issue of stopping dialysis. Nearly 70% of patients were incompetent at the time of the decision. Patient refusal to stop dialysis was uncommon. CONCLUSIONS: Stopping dialysis before death is a common practice in our unit. Dialysis was mostly discontinued in severely ill patients who were near the end of their life. The nephrologist or the renal team decided to stop treatment. Our results should encourage renal teams to raise the issue of stopping dialysis when a patient's illness has become terminal. More studies and discussion of this difficult field are needed. PMID- 16257846 TI - Elderly patients on haemodialysis have 50% less functional capacity than gender- and age-matched healthy subjects. AB - OBJECTIVE: Patients on haemodialysis (HD) are by necessity inactive for 3-6 h three times a week. The aim of this prospective controlled study was to investigate the level of functional capacity of elderly HD patients using simple measures. MATERIAL AND METHODS: A total of 11 consecutive HD patients aged > 60 years (median age 75 years) were included in the study. They were compared with 22 gender- and age-matched healthy subjects using a prospective 1:2 case:healthy subject study design. As tests of functional capacity and maximal exercise capacity we used a "sit-to-stand" test (number of cycles within 10 s) and a staircase test (the number of cycles completed per second was used as the effect variable), respectively. RESULTS: The patients managed significantly fewer cycles than the healthy subjects in the staircase test (median 2 vs 10; p<0.003) and performed approximately 50% fewer cycles in the sit-to-stand test (p<0.014). The work performed in a given time in the staircase test was 54% less for the patients (p<0.017). CONCLUSIONS: The HD patients in the study had a considerably lower functional capacity than the healthy subjects, which may have interfered with their daily living activities. The tests used are easy to apply and need no specialized equipment. The importance of investigating functional capacity and instituting rehabilitation programmes is emphasized. PMID- 16257847 TI - Improvement of oligoasthenozoospermia in epileptic patients on switching anti epilepsy medication from sodium valproate to phenytoin. AB - We present two cases of infertile male patients with oligoasthenozoospermia who were receiving anti-epilepsy medication. Complete reversal of the spermatic dysfunction followed by a successful conception was achieved after discontinuation of sodium valproate, suggesting that the drug was responsible for spermatic dysfunction in these individuals. PMID- 16257848 TI - Investigations of clofibrate in alternative carcinogenicity models. PMID- 16257849 TI - Evaluation of the carcinogenic potential of clofibrate in the p53+/- mouse. AB - This study was conducted as part of International Life Sciences Institute (ILSI) program to evaluate the carcinogenic potential of clofibrate, a nongenotoxic, peroxisome proliferator-activated receptor (PPAR) alpha agonist, following oral administration to p53+/- heterozygous mice for a minimum of 26 weeks. p Cresidine, a urinary bladder carcinogen, was given orally at 400 mg/kg/day as a positive control. Initial clofibrate doses were 50, 250, and 400 mg/kg/day for males and 50, 200, and 500 mg/kg/day for females. Due to unexpected mortality during the first week of dosing, clofibrate doses were lowered to 25, 75, and 100 mg/kg/day for males and 25, 75, and 125 mg/kg/day for females. Clinical signs and mortality were greater in p53+/- than wild-type (WT) mice. With the exception of liver weights, no marked differences in any other parameters either between the sexes or between WT and p53+/- mice were noted. Moderate increases in liver weights noted in WT males given 100 mg/kg/day clofibrate were not associated with any microscopic changes. No neoplastic response was observed in p53+/- mice after 6 months of exposure to clofibrate at doses up to 100 mg/kg/day for males and 125 mg/kg/day for females. Transitional-cell hyperplasia and carcinoma of the urinary bladder were noted in both sexes given p-cresidine, demonstrating that the p53+/- mouse responded to a known mouse carcinogen as expected. Clofibrate produced non neoplastic findings in the adrenals, pancreas, and prostate, whereas p-cresidine affected the kidney, liver, pancreas, and spleen. PMID- 16257850 TI - Evaluation of the carcinogenic potential of clofibrate in the rasH2 mouse. AB - The purpose of the study was to support of the International Life Sciences Institute (ILSI) alternative carcinogenicity models initiative to evaluate the carcinogenic potential of the nongenotoxic carcinogen, clofibrate, a peroxisome proliferator-activated receptor (PPAR) alpha agonist, following oral administration to rasH2 mice. Peroxisome proliferators are one of the most widely studied of the nongenotoxic carcinogens and have diverse industrial and therapeutic uses (Gonzalez et al. J. Nat. Cancer Inst. 90: 1702-1709, 1998); however, the nongenotoxic mechanism of carcinogenicity is currently unknown. Male mice were administered doses of clofibrate at 50, 100, or 200 mg/kg/day and female mice were administered doses of 50, 150, or 250 mg/kg/day by oral gavage at 10 ml/kg for 27 weeks. In addition, rasH2 male and female mice were treated with N-nitroso-N-methylurea (NMU). Nontransgenic male and female mice were treated with 200 and 250 mg/kg/day, respectively, of clofibrate. The NMU-treated mice were given a single intraperitoneal dose of 75 mg/kg, which was followed by a 90-day observation period; all others were sacrificed after 6 months of daily dosing. Hepatocellular neoplasms were observed in clofibrate-treated rasH2 male mice after 6 months of treatment but not in nontransgenic males or females. Clofibrate treatment (250 mg/kg/day) of female rasH2 mice was associated with a slight increase in the incidence of various neoplasms (harderian gland, lungs, skin, spleen, tail, thymus, and uterus) compared with untreated transgenic mice and with similarly treated nontransgenic mice. Non-neoplastic changes were found in the liver of transgenic and nontransgenic mice of both sexes and in the kidneys of male mice. NMU produced findings are consistent with previous studies. The data suggest that the rasH2 mice are a good model for testing epigenetic carcinogens in a shorter timeframe than conventional mouse carcinogenicity bioassays. PMID- 16257852 TI - Evaluation of the carcinogenic potential of clofibrate in the FVB/Tg.AC mouse after dermal application--part II. AB - This study was conducted as part of the International Life Sciences Institute (ILSI) Alternatives to Carcinogenicity Testing program and evaluated the carcinogenic potential of clofibrate, a nongenotoxic, peroxisome proliferator activated receptor (PPAR) alpha agonist following dermal application to transgenic Tg.AC and nontransgenic FVB mice for a minimum of 26 weeks. Clofibrate doses of 12, 28, or 36 mg/200 microl/day were used. Positive controls for papilloma formation were benzene (174.8 mg/200 microl), and 12-o tetradecanoylphorbol-13-acetate (TPA [0.00250 mg/200 microl]). Clofibrate was tolerated at doses up to 36 mg/200 microl. In Tg.AC mice, clofibrate produced a dose-related increase in the incidence of mice with cutaneous papillomas; and dose-related decreases in mean time to first tumor, mean multiplicity of tumors per mouse, and mean weeks to maximal yield, as well as numerous nonneoplastic microscopic lesions in the liver, kidney, spleen, and skin. Benzene and TPA induced both neoplastic and/or non-neoplastic proliferative lesions in Tg.AC mice. Clofibrate did not increase the incidence or multiplicity of papillomas, or any other tumors in FVB mice. These data show that the Tg.AC dermal model has increased sensitivity in detecting skin papillomas caused by the nongenotoxic rodent carcinogen, clofibrate, compared to wild type FVB mice, at systemic exposures that are 3x higher than the systemic exposure observed in humans taking clofibrate (AUC = 1100 microg.h/ml) at the recommended maximum therapeutic dose of 500 mg. In addition, this study supports the proposed concept that Tg.AC model may detect compounds with nongenotoxic carcinogenic potential in a shorter timeframe than conventional mouse carcinogenicity bioassays. PMID- 16257851 TI - Evaluation of the carcinogenic potential of clofibrate in the FVB/Tg.AC mouse after oral administration--part I. AB - This study was conducted as part of the International Life Sciences Institute (ILSI) program to evaluate the carcinogenic potential of clofibrate, a nongenotoxic, peroxisome proliferator-activated receptor (PPAR) alpha agonist following oral administration to Tg.AC (transgenic) and wild-type FVB (nontransgenic) mice for a minimum for 6 months. Clofibrate was well tolerated at doses up to 500 (males) and 650 (females) mg/kg/day. Oral administration of clofibrate to Tg.AC or FVB (wild-type) male and female mice for 6 months did not result in the increased formation of neoplastic lesions. Epithelial hyperplasia in the urinary bladder (Tg.AC and FVB) and prostate gland (Tg.AC only), and interstitial-cell hyperplasia in the testes (Tg.AC) were noted at 500 mg/kg/day. Non-neoplastic nonproliferative findings included hepatic hypertrophy and hematopoietic changes (myeloid hyperplasia, myelodysplasia, lymphoid depletion, and erythropoiesis) in Tg.AC and FVB mice of both sexes; reproductive (cystic degeneration and dilatation, hypospermia, spermatocele, dilated inspissated protein) and urogenital (tubular-cell hypertrophy, degenerative/regenerative nephropathy, necrosis/fibrosis) changes in Tg.AC and FVB male mice; congestion in the lung in male Tg.AC mice; gall bladder dilatation in female Tg.AC mice; and adrenal (intracellular lipofuscinosis and atrophy) and heart (eosinophillic myofibers) findings in Tg.AC mice of both sexes and in female FVB mice. The results of this study indicate that the clofibrate is not carcinogenic when administered to Tg.AC mice by oral gavage for 6 months at doses up to 500 (males) and 650 (females) mg/kg/day, which did produce liver hypertrophy. PMID- 16257853 TI - Evaluation of the carcinogenic potential of clofibrate in the neonatal mouse. AB - This study was conducted in support of the International Life Sciences Institute (ILSI) alternative carcinogenicity models initiative to evaluate the carcinogenic potential of clofibrate, a nongenotoxic peroxisome proliferator-activated receptor (PPAR) alpha agonist, following oral administration to neonatal mice. Male and female neonatal CD-1 mice were dosed with clofibrate at doses of 100, 250, and 500 mg/kg or with the positive control, diethylnitrosamine (DEN), at 2 mg/kg by oral gavage on days 9 and 16 post birth and observed for approximately 1 year for the development of tumors. Plasma levels of clofibric acid after the second administration increased with dose, but were not dose proportional. Clofibrate administered by gavage on litter days 9 and 16 to neonatal mice at doses of 100, 250, or 500 mg/kg did not produce a carcinogenic effect. The positive control DEN did produce tumors in the liver and lung (single and multiple adenomas and carcinomas) and harderian gland (adenoma) of both sexes. Non-neoplastic lesions related to DEN treatment were confined to myocardial degeneration/fibrosis and testicular interstitial hyperplasia in males, and to glomerulonephrosis and gastritis in both sexes. PMID- 16257854 TI - Studies evaluating the utility of N-methyl-N-nitrosourea as a positive control in carcinogenicity studies in the p53+/- mouse. AB - Studies conducted under the auspices of International Life Sciences Institute (ILSI) have suggested that an alternative mouse carcinogenicity study may be substituted for the traditional 2-year mouse bioassay typically conducted to support the development of drug candidates. The purpose of this study was to characterize the carcinogenic potential of N-methyl-N-nitrosourea (MNU), a DNA alkylating agent, in p53+/- knockout mice to determine its suitability as a positive control agent in an alternative carcinogenicity model. p53+/- knockout mice were administered a single oral dose of 90 mg/kg and maintained for up to 13 weeks prior to evaluation of neoplasms. Treatment was generally well tolerated; however, 4 of 30 mice died between the days of 75 and 92 due to neoplasms. MNU related macroscopic observations included enlargement of the thymus, spleen, mandibular and mesenteric lymph nodes; and pale liver, heart, kidney, and bone marrow, which correlated with the diagnosis of lymphoma of the hematopoietic system, noted in the thymus of all affected animals and in the spleen, liver, lungs, and kidneys of some animals. Other treatment-related single neoplasms included a squamous-cell carcinoma in the nonglandular stomach and leiomyosarcoma in the glandular stomach. Non-neoplastic proliferative lesions included acanthosis and hyperkeratosis in the nonglandular stomach, focal papillary hyperplasia of the nonglandular stomach, glandular hyperplasia of the stomach, and adenomatous hyperplasia of the duodenum or ileum. The increased incidence of neoplastic and proliferative changes in MNU-treated mice suggests MNU could serve as a positive control in alternative carcinogenicity studies conducted in p53+/- knockout mice. PMID- 16257855 TI - Concomitant administration of bevacizumab, irinotecan, 5-fluorouracil, and leucovorin: nonclinical safety and pharmacokinetics. AB - Bevacizumab (Avastin) is a humanized monoclonal antibody against vascular endothelial growth factor approved for use in combination with 5-fluorouracil (5 FU)-based chemotherapy for first-line treatment of metastatic colorectal cancer. The Saltz regimen (irinotecan/5-FU/leucovorin [LV]) is a first-line treatment for this indication. The objective of this study was to evaluate the safety of bevacizumab when administered concomitantly with the Saltz regimen to cynomolgus monkeys, and to determine if the pharmacokinetics of bevacizumab, irinotecan, SN38 (the active metabolite of irinotecan), or 5-FU were affected by combined administration. Male cynomolgus monkeys were intravenously administered the Saltz regimen (125 mg/m2 irinotecan, 500 mg/m2 5-FU, 20 mg/m2 LV) alone (n = 4) or concomitantly with 10 mg/kg bevacizumab (n = 5) on days 1 and 8. All animals survived to euthanasia on day 15. Adverse effects associated with the Saltz regimen included diarrhea and neutropenia. Macroscopically, two animals from each group had small thymus glands that correlated microscopically with lymphoid depletion. Myeloid hypoplasia and/or erythroid hyperplasia was observed in the sternal bone marrow of most animals. These effects were considered to be associated with the Saltz regimen; concomitant bevacizumab administration did not alter the severity of these findings. Irinotecan and 5-FU were observed to be rapidly eliminated (t1/2 = 1 h and 0.5 h, respectively). Although the number of animals in each group was small and no statistical comparison between groups was performed, bevacizumab did not affect the disposition of either agent. These results indicate that bevacizumab can be safely administered in combination with the Saltz regimen without pharmacokinetic interaction. PMID- 16257856 TI - Effect of dietary fat on selected parameters of toxicity following 1- or 3-month exposure of rats to toluidine isomers. AB - The aim of the study was to investigate the effect of the dietary fat on selected parameters of toluidines toxicity in rats during subchronic exposure. Three isomers of toluidine (ortho, meta, and para) were administered to rats in the diet for 1 and 3 months at levels 40, 80, 160 mg/kg/day in two kinds of diet containing either 4% or 14% fat. All doses of toluidine isomers produced a 1.5- to 9.8-fold increase in methemoglobin (MetHb) level during both treatment periods. A distinct dose-response relationship was observed, especially for o- and m-toluidine; the effect was generally greater in rats fed high-fat diet. Reduced glutathione level in liver was increased in all treated groups, 1.5- to 5.1-fold, irrespective of the kind of diet. An increase in hepatic lipid peroxidation (thiobarbituric acid reactive substances; TBARS), 1.5- to 4.5-fold, was noticed in the majority of the treated groups. Generally, there was no consistent effect of diet except for p-toluidine where the level of hepatic TBARS was lower in rats fed high-fat diet. Blood urea nitrogen (BUN) level in animals treated with all doses of o- and m-toluidine was 1.3- to 5.0-fold higher in comparison with respective controls. No clear relationship between BUN level and the kind of diet was found. No effect of toluidines on the activity of serum aspartate aminotransferase (AST) and sorbitol dehydrogenase (SDH) were observed. In the majority of groups treated for 30 and 90 days the amount of toluidines in 24-h urine was lower in rats fed high-fat diet. Final body weight gain in rats treated with o- and p-toluidine (80 and 160 mg/kg body weight [b.w.]) was lower as compared to controls. In summary the high-fat diet stimulated methemoglobin formation in rats treated with o- and m-toluidine and cause the decrease in the amount of toluidines in 24-h urine. The high content of fat did not affect consistently the other parameters tested. PMID- 16257857 TI - Individual reference intervals of hematological and serum biochemical parameters in cynomolgus monkeys. AB - Cynomolgus monkeys, one of a number of primates phylogenetically close to humans, are commonly used in animal studies. The purpose of this study was to assess biological variations in hematological and serum biochemical parameters in cynomolgus monkeys. Summary statistics and reference intervals were calculated using data from 95 male and 95 female Chinese-bred cynomolgus monkeys aged 3 to 7 years showing no abnormalities during the breeding period. Within- and between animal variations were estimated using a random-effect analysis of variance (ANOVA), then, a simple method that applies prior information was proposed to estimate individual reference intervals. Parameters including MCV, MCH, PT, ALP, total cholesterol, and creatinine appeared to show a large between-animal variation; thus, it is considered that individual reference intervals for these parameters would be relatively small in comparison with overall reference intervals. PMID- 16257859 TI - Autophagic degeneration of motor neurons in a model of slow glutamate excitotoxicity in vitro. AB - There is increasing evidence that so-called "autophagic cell death" participates in cell degeneration in certain pathological conditions. Autophagy might be involved in some neurodegenerative processes, including lateral amyotrophic sclerosis (SLA). The exact mechanism leading to progressive motor neuron (MN) loss remains unclear, but glutamate-mediated mechanism is thought to be responsible. Previous ultrastructural studies by the authors performed on a model of SLA in vitro, based on chronic glutamate excitotoxicity, revealed a subset of morphological features characteristic to different modes of neuronal death, including autophagic degeneration. The contribution of this pathway of MNs death is evaluated in organotypic cultures of rat lumbar spinal cord chronically exposed to specific glutamate uptake blockers: DL-threo-beta-hydroxyaspartate (THA) and L-transpyrrolidine-2,4-dicarboxylate (PDC). The study documents the various steps of authophagy in slowly evolving process of MN neurodegeneration. The cells undergoing autophagy usually exhibited sequestration of some parts of cytoplasm with normal and/or degenerated organelles, whereas other parts of cytoplasm as well as neuronal nucleus remained unchanged. The advanced autophagic changes were often associated with other modes of MN death, especially with apoptosis. Numerous MNs revealed apoptotic nuclear features with typical peripheral margination of nuclear chromatin, accompanied by severe autophagic or autophagic-necrotic degeneration of the cytoplasm. These results support the opinion of unclear distinction between different modes of cell death and indicate the involvement of autophagey in MNs neurodegeneration in vitro. PMID- 16257860 TI - The ultrastructure of hybrid acute leukemia: a study of 15 cases. AB - The objective of this study was to investigate the ultrastructural characteristics of hybrid acute leukemia (HAL). Fifteen cases of HAL were studied by transmission electron microscopy (TEM), focusing on organelles and myeloperoxidase (MPO) reaction of leukemic cells. By TEM, 5 out 15 cases of HAL were consistent with immunophenotyping (3 cases of biphenotypic type, and 2 cases of biclonal type with granulocytes and lymphocytes); 2 cases were suspected as HAL. On other hand, 5 cases of HAL were assigned to ALL, and 2 cases were misinterpreted as M5a and 1 as M4b. Most of the blast cells of biphenotypic HAL showed lymphoid features, except some cases containing MPO positive granules in blasts, while a few cases exhibited monocytic or nonspecific features. TEM offers advantages in the diagnosis of biclonal type HAL and biphenotypic HAL positive for MPO. However, it is difficult to differentiate MPO-negative cases of biphenotypic HAL from ALL and a few cases may be misinterpreted as M5 by TEM. PMID- 16257861 TI - Morphometric study of desmosomes from oral squamous cell carcinoma. AB - The authors studied ultrastructural characteristics of desmosomes from oral squamous cell carcinoma, reporting the cellular differentiation and size of the desmosomes. The length of the desmosome profiles was measured with a Zeiss KS 300. The desmosomes were grouped according to their size and the tumor histological grading. Statistical analysis indicated a significant correlation (p < .001) between the size of the desmosomes and the histological grading group of the malignancy. The comparison of the desmosome size among the tumor histological grading groups also showed significant difference (p < .001). A multiple comparisons test indicated homogeneity in the size of desmosomes within the histological grading groups: 100% in the well differentiated, 95.2% in the moderately differentiated, and 50% in the poorly differentiated group. The preliminary data strongly suggest that the homogeneity of length of the desmosome profiles may be exploited for diagnostic strategies. PMID- 16257862 TI - Tubuloreticular structures in rectal biopsies of SIV-infected rhesus monkeys (Macaca mulatta). AB - Tubuloreticular structures (TRS) are considered to be a specific ultrastructural marker for AIDS in various organs. Experimental SIV infection in rhesus macaques is the most appropriate animal model of HIV infection. In 8 rhesus monkeys, experimentally infected with SIVmac251/MPBC, rectum biopsies were taken prior to and post infection (day 3; 1, 2, 4, 12 weeks p.i.) and were investigated by transmissionelectron microscopy to determine incidence and extent of tubuloreticular structures as well as affected cells. From the first week p.i. on TRS were found in all experimental animals as tubuli with a diameter of 20-30 nm. The tubuli were arranged in regular paracristalline formations and formed intracytoplasmatic heterogenous, polymorph accumulations, which were localized close to the endoplasmatic reticulum. In the rectal lamina propria macrophages, endothelial cells, plasma cells, lymphocytes, fibroblasts, and neutrophilic granulocytes were the affected cell types. In 5 control biopsies TRS were detected, too, but, in contrast to SIV-infected animals, they appeared only singular and very small. The results indicate that TRS are a characteristic morphologic criteria of intestinal SIV infection. They appear in very early stages of the infection. In the rectum, they can be detected as bigger, conspicuous, and abundant formations in several cells and have a restricted diagnostic and prognostic validity. PMID- 16257864 TI - Ultrastructural studies in a series of 18 cases of chromophobe renal cell carcinoma. AB - Chromophobe renal cell carcinoma (CRCC) is a distinct variant of renal epithelial neoplasm. Ultrastructurally, the presence of numerous cytoplasmic microvesicles is highly characteristic of CRCC, as are mitochondria with tubulovesicular cristae. All 18 cases is this study present these two elements with a different distribution, depending on the morphological variant (typical or eosinophilic). In both variants, the mitochondria show tubulovesicular cristae and budding from the outer mitochondrial membrane that is very similar to the cytoplasmic microvesicles. An association was also found between intracristal mitochondrial swelling and bud formation. These buds, when detached from the mitochondrial membrane, may convert into the cytoplasmic microvesicles. PMID- 16257863 TI - Immunohistochemical and ultrastructural features of neuroendocrine differentiated carcinomas of the prostate: an immunoelectron microscopic study. AB - The purpose of this study was to further define the immunohistochemical and ultrastructural characteristics of neuroendocrine (NE) differentiated prostatic carcinomas. Seventy-seven specimens were obtained from prostatic carcinoma tumors during prostatectomy, transurethral resection of prostate or biopsy in 77 prostate cancer patients, and analyzed by immunohistochemical staining for chromogranin A (CgA). Nine of these tumors were also studied by elctron microscopy and 4 were examined by pre-embedding immunoelectron microscopy. CgA stained cells were detected in 36 tumors (47%). Clinically advanced tumors or tumors with higher histological grades were associated with increased NE differentiation. Three of the tumors studied by electron microscopy contained cells showing unequivocal NE differentiation revealed by the presence of neurosecretory granules, while the poorly NE-differentiated malignant cells contained pleomorphic granules, which were lysosomal-like rather than NE-type granules. Immunoelectron microscopy demonstrated the presence of CgA immunoreactivity on the pleomorphic granules in the poorly differentiated malignant glands. This study suggests that NE-differentiated malignant cells in prostate cancer tissues may induce aggressive behavior in adjacent proliferating neoplastic cells via a paracrine mechanism. PMID- 16257866 TI - Bilateral oncocytic malignant melanoma in axillary lymph nodes without evidence of an extranodal primary. AB - A malignant melanoma was diagnosed in an axillary lymph node of a 49-year-old man. The tumor was examined by electron microscopy and was found to be composed of large oncocytic cells, filled with abundant mitochondria. No primary tumor could be identified on the skin or within internal organs. Approximately 2 years after the initial diagnosis, the patient presented with malignant melanoma in an axillary lymph node on the contralateral side. The second tumor also expressed the same oncocytic phenotype, favoring the common origin of both tumors. These data illustrate that oncocytic melanomas may retain their oncocytic phenotype during metastatic dissemination. PMID- 16257865 TI - Primary smooth muscle tumor of the pleura: a clinicopathological case report with ultrastructural observations and a review of the literature. AB - Primary smooth muscle tumor of the pleura is exceptionally rare. The authors describe a primary smooth muscle tumor of the pleura that was discovered incidentally on chest X-ray in a 73-year-old man. Magnetic resonance imaging demonstrated a 12 x 18 x 15-cm pleura-based mass arising from the posterior mediastinum. Computerized tomography (CT) guided needle cores from the pleura showed a primary smooth muscle tumor of undetermined malignant potential. Further excision of the whole tumor showed an intimate relation to pleura, and the diagnosis of leiomyosarcoma was made. The clinical, radiological, histopathological, immunohistochemical, and ultrastuctural findings were consistent with a primary smooth muscle tumor of the pleura. This is the seventh case in the literature of a primary smooth muscle tumor of the pleura, which, to the best of the authors' knowledge, is the first such case of the pleura to be diagnosed on CT-guided needle biopsy. In conclusion, this method of investigation is recommended since it is minimally invasive but has a rewarding yield in providing the most likely diagnosis, predicting prognosis, and management planning. PMID- 16257867 TI - Chronic inflammation and pain inside the mandibular jaw and a 10-year forgotten amalgam filling in an alveolar cavity of an extracted molar tooth. AB - A 55-year-old woman, suffered from severe pain in her mandibular jaw for several years. A metallic artifact of about 2(3) mm was detected by a panorama radiography in an edentulous region with a surrounding inflammation in close contact to the canal of the mandibular nerve. Inflammated tissue with the central metallic inclusion was removed from the bone under local anesthesia and operation. Postoperatively, pain and missensitivity disappeared within 1 week. Although the patient had no macroscopically visible so-called amalgam tattoo, the metallic cube was identified as amalgam by the detection of mercury, silver, tin, copper, and zinc using energy dispersive X-ray analysis (EDX) in a scanning electron microscope (SEM). Nevertheless, brown to black pigments in the connective tissue matrix and inside histiocytes, fibroblasts, and multinucleated foreign giant cells of the surrounding inflammatory tissue were observed by light and electron microscopy. However, the elemental analysis by EDX in SEM or by electron energy loss spectroscopy in transmission electron microscope detected only silver, tin, and sulfur but no mercury in these precipitates and in the residual bodies of phagocytes. The presented case demonstrates a seldom complication of amalgam deposition in the tissue. The authors assume that the chronic pain results from a forgotten amalgam filling inside an alveole after extraction of a molar tooth, causing a chronic inflammation by resolving mercury and other toxic elements out of the metallic artifact. PMID- 16257868 TI - Asbestos burden in cases of mesothelioma from individuals from various regions of the United States. AB - Mesothelioma is a rare tumor that is considered an asbestos marker disease. It occurs in individuals following a longer latency period from first exposure than other asbestos-related diseases. The tumor also occurs in individuals with a wide range of exposures, including individuals with lower level or secondary exposures. In the present study lung tissue from 54 individuals with a pathological diagnosis of mesothelioma was evaluated for ferruginous body and uncoated asbestos fiber content. The data were compared with an earlier study of mesothelioma cases from the northwestern United States. Tissue was prepared via a digestion procedure, with the collected digestate reviewed by light microscopy for quantification of asbestos bodies and analytical transmission electron microscopy for determination of uncoated fiber burden. Twenty-seven cases in the present study had over 1000 ferruginous bodies per gram of dry tissue. The data suggest that amosite provides a more likely stimulus for ferruginous coating than the other forms of asbestos. All individuals were found to have asbestos fibers in their lung tissue. Amosite was the most commonly found fiber, with anthophyllite being the second most commonly found type of asbestos. The finding of tremolite in the tissue most often was associated with the finding of anthophyllite. A limited number of asbestos fibers of each type would have been seen in the light microscope, with the least detected being chrysotile. The majority of all fiber types were found as short fibers (< 8 mum), although some longer fibers were represented in each type of asbestos. The majority of the individuals were found to have mixed types of asbestos in their lungs. PMID- 16257869 TI - Hypertension and coronary artery disease: mechanistic insights and therapeutic challenges. PMID- 16257870 TI - ASCOT - more than a horse race! PMID- 16257871 TI - The association of hypertension and aortic valve sclerosis. AB - BACKGROUND: Aortic valve sclerosis (AVS), a condition of thickening and calcification of the normal trileaflet aortic valve without the obstruction to left ventricular outflow, is likely the initial stage in the development of aortic stenosis and is associated with an increased incidence of cardiovascular events. The objective of this study is to critically review the data on the association of blood pressure and hypertension with AVS. METHODS: A systematic search of MEDLINE and EMBASE (to June 2004) was conducted using the keywords hypertension and aortic valve. All English language papers were examined if they dealt with hypertension and AVS. All studies were included for analysis if they had a control group. RESULTS: Three population-based, cross-sectional studies with a total sample size of 6450 individuals showed a consistent and significant relationship between hypertension and AVS with an odds ratio (OR) ranging from 1.23 to 1.74. Smaller case-control studies with a total sample size of 1609 individuals did not show consistent results but the OR ranged from 1.75 to 2.38. Only one small study (n = 188) showed fewer cases with hypertension and AVS than in the control group. Hypertension was a significant factor remaining in multivariate analysis after consideration of age and other risk factors in several cross-sectional studies. In contrast, other studies with blood pressure measurements consistently showed no increased blood pressures in the presence of AVS. However, these studies did not examine the prevalence of AVS within age adjusted blood pressure levels. CONCLUSIONS: Cross-sectional population-based studies present evidence of an association between hypertension and AVS with an OR between 1.23 and 1.74. The major limitation in establishing a causal relationship is the failure to demonstrate a gradient of risk between increasing blood pressure and increasing incidence of AVS. In addition, the literature is confounded by the wide variety of definitions for AVS as well as hypertension. At this time, further data is required to conclude that there is a causal relationship between AVS and elevated blood pressure. PMID- 16257872 TI - Influence of regression of left ventricular hypertrophy on left atrial size and function in patients with moderate hypertension. AB - OBJECTIVES: The aim of the study was to evaluate the effect of regression of left ventricular (LV) hypertrophy on left atrial (LA) size and function in patients treated with telmisartan, an angiotensin II receptor blocker. METHODS: Patients population included 80 patients with mild-moderate LV hypertrophy treated with telmisartan. Patients were followed over a period of 12 months from the start of telmisartan treatment. LA size was measured during systole from the parasternal long-axis view from M-mode. Atrial function was assessed by Doppler echocardiography and the following parameters were measured: transmitral peak A velocity, atrial filling fraction, atrial ejection force (AEF), peak E velocity, deceleration time and isovolumic relaxation time, LA maximal and minimal volume, and LV cardiac mass index (LVMI). RESULTS: All patients had an increased LVMI and decrease during follow-up. LA dimensions were greater at baseline and reduced after 1 year of treatment. LA volume indexes maximal volume, minimal volume and P volume were reduced compared with baseline value (maximal volume from 35+/-5 to 32+/-5, p<0.05; minimal volumes from 14+/-2 to 10+/-4, p<0.05). AEF, a parameter of atrial systolic function, increased from 12+/-3 to 15+/-2.4 (p<0.01). The reduction of LA volumes correlate with reduction of LVMI (LA maximal volume and LVMI r = 0.45; p<0.01; LA minimal volume and LVMI r = 0.34; p<0.05). A positive correlation was also found between LV mass index and P volume (r = 0.41; p<0.01), LV mass index and LA active emptying volume (r = 0.39; p<0.01), and LV mass index and LA total emptying volume (r = 0.38; p<0.05). CONCLUSIONS: The present study suggests that regression of LV hypertrophy due to telmisartan is associated with reduction of LA volumes that expresses variation of LV end-diastolic pressure. The reduction of LV end-diastolic pressure is associated with an increase in diastolic filling and with a significant reduction of active and passive emptying contribution of left atrium to LV stroke volume. PMID- 16257873 TI - Plasma endothelin-1 is not increased in overweight/obese hypertensive African women. AB - INTRODUCTION: Endothelin-1 (ET-1) has been found to be higher in hypertensive African Americans and obese hypertensive Caucasians compared to normotensive controls with an enhanced ET-1-dependent vasoconstrictor tone. ET-1 levels and the associations thereof with cardiovascular function in overweight/obese normotensive and hypertensive African women have not been investigated. It is therefore hypothesized that ET-1 levels are elevated in overweight/obese hypertensive African women compared to overweight/obese and lean normotensive controls. Additionally, it is hypothesized that these elevated ET-1 levels are associated with increased total peripheral resistance (TPR) and decreased arterial compliance (C(W)). MATERIALS AND METHODS: A case-case control study was performed which included 98 African women. The subjects were divided into lean normotensive (lean NT), overweight/obese normotensive (OW/OB NT) and overweight/obese hypertensive (OW/OB HT). The Finometer apparatus was used to obtain a more elaborate cardiovascular profile and plasma immunoreactive ET-1 levels were determined. RESULTS: ET-1 levels were similar for the three groups. Although a decrease in vascular function was observed in the OW/OB HT group, no correlations were obtained between ET-1 and the cardiovascular profile, before and after adjusting for age. CONCLUSION: In African women, ET-1 levels did not differ between lean and overweight/obese and normotensive and hypertensive subjects. The lack of significant associations between ET-1 and decreased vascular function in the overweight/obese hypertensive group suggests that ET-1 is not implicated in obesity-related hypertension in African women. PMID- 16257874 TI - Determinants of poor hypertension management in patients with ischaemic heart disease. AB - Not much is known about factors influencing hypertension management in patients with ischaemic heart disease (IHD). Therefore, the aim of the study was to assess factors influencing hypertension management in patients hospitalized due to IHD. We reviewed hospital records of 1051 consecutive patients with a discharge diagnosis of myocardial infarction (MI; n = 290), unstable angina (n = 247), percutaneous coronary intervention (PCI; n = 259) or coronary artery bypass grafting (CABG; n = 255) who were hospitalized at three university (n = 533) or three community (n = 518) cardiac departments. During the follow-up interview (6 18 months after discharge) 70.2% of study participants fulfilled the criteria for a diagnosis of hypertension. Hypertension had not been diagnosed during index hospitalization in 17.5% of hypertensive participants. Overall, 7.1% of hypertensives were not treated with any blood pressure lowering agent. Irregular health checks (odds ratio, OR, 16.3, 95% confidence interval, CI, 4.1-64.0), alcohol drinking (OR 3.3, 95% CI 1.5-7.0), unstable angina (OR 2.7, 95% CI 1.3 5.8), hypertension awareness (OR 0.2, 95% CI 0.1-0.5) and blood pressure lowering drugs prescribed at discharge (OR 0.08, 95% CI 0.03-0.19) were significantly related to the probability of not being on antihypertensive medication. High blood pressure (>or=140/90 mmHg) was found in 68.9% of hypertensives; older age (OR 1.3, 95% CI 1.0-1.6) and hypertension awareness (OR 0.6, 95% 0.3-1.0) were the only significant predictors of uncontrolled hypertension. Among treated participants with uncontrolled hypertension, 33.4% were on monotherapy, 66.6% were on combination therapy, 25.5% were on three or more drugs and 14.7% were on combination of three or more drugs with diuretic. CONCLUSIONS: Hypertension management in the secondary prevention of IHD is not satisfactory. Age and hypertension awareness are the main factors related to the quality of blood pressure control in the post-discharge period. PMID- 16257875 TI - Aortic pulse pressure and aortic pulsatility are associated with angiographic coronary artery disease in women. AB - BACKGROUND: Studies indicated that both aortic pulse pressure (PP) and aortic pulsatility are independently associated with angiographic coronary artery disease (CAD). However, most of these studies included a majority of male subjects, and women were underrepresented. OBJECTIVE: We investigated the relation of aortic PP and aortic pulsatility derived from invasively measured ascending aortic pressure waveform and presence of angiographic CAD in women undergoing diagnostic coronary angiography. METHODS AND RESULTS: From September 2003 to April 2004, 262 unselected female subjects undergoing first cardiac catheterization were consecutively included in the study. Systolic, diastolic and mean pressure waveforms of the ascending aorta were measured during cardiac catheterization with a fluid-filled system. Aortic pulsatility was estimated as the ratio of aortic PP to mean pressure. Angiographic CAD was detected in 175 (67%) patients. In multiple-adjusted logistic regression, both aortic PP and aortic pulsatility were significantly associated with the presence of CAD (for a 10-mmHg increase in PP: odds ratio [OR] = 1.3, 95% confidence interval [CI] = 1.1 1.76; for a 0.1 increase in aortic pulsatility: OR = 1.8, 95% CI = 1.3-2.4). When patients were divided into tertiles according to the level of aortic pulsatility, it was noted that multiple-adjusted OR of presence CAD was 2.2 (95% CI = 1.1-4.4) for the middle tertile of the aortic pulsatility level and 5.9 (95% CI = 2.7 12.8) for the highest tertile of the aortic pulsatility level compared with the lowest tertile. CONCLUSION: In female subjects referred to coronary angiography, ascending aorta PP and aortic pulsatility are significantly associated with the presence of angiographic CAD and these associations are independent of age and other cardiovascular risk factors. PMID- 16257877 TI - Comparison of randomization techniques for clinical trials with data from the HOMERUS-trial. AB - BACKGROUND: Several methods of randomization are available to create comparable intervention groups in a study. In the HOMERUS-trial, we compared the minimization procedure with a stratified and a non-stratified method of randomization in order to test which one is most appropriate for use in clinical hypertension trials. A second objective of this article was to describe the baseline characteristics of the HOMERUS-trial. METHODS: The HOMERUS population consisted of 459 mild-to-moderate hypertensive subjects (54% males) with a mean age of 55 years. These patients were prospectively randomized with the minimization method to either the office pressure (OP) group, where antihypertensive treatment was based on office blood pressure (BP) values, or to the self-pressure (SP) group, where treatment was based on self-measured BP values. Minimization was compared with two other randomization methods, which were performed post-hoc: (i) non-stratified randomization with four permuted blocks, and (ii) stratified randomization with four permuted blocks and 16 strata. In addition, several factors that could influence outcome were investigated for their effect on BP by 24-h ambulatory blood pressure monitoring (ABPM). RESULTS: Minimization and stratified randomization did not lead to significant differences in 24-h ABPM values between the two treatment groups. Non stratified randomization resulted in a significant difference in 24-h diastolic ABPM between the groups. Factors that caused significant differences in 24-h ABPM values were: region, centre of patient recruitment, age, gender, microalbuminuria, left ventricular hypertrophy and obesity. CONCLUSION: Minimization and stratified randomization are appropriate methods for use in clinical trials. Many outcome factors should be taken into account for their potential influence on BP levels. Recommendation. Due to the large number of potential outcome factors that can influence BP levels, minimization should be the preferred method for use in clinical hypertension trials, as it has the potential to randomize more outcome factors than stratified randomization. PMID- 16257876 TI - Isolated office hypertension: a 3-year follow-up study. AB - The study aimed to evaluate, over a 3-year period, the progression towards sustained hypertension and left ventricular (LV) changes in patients with isolated office (IO) hypertension (office BP>140 and/or 90 mmHg, daytime BP<130/80 mmHg). After 3 years from the basal evaluation, 38 subjects with basal normal BP and 42 subjects with basal IO hypertension underwent a second 24-h BP monitoring and echocardiography; 19 patients of the basal IO hypertension group were not revaluated because they had already developed ambulatory hypertension and were on antihypertensive treatment. At the second evaluation, the 38 normotensive subjects had unchanged BP and LV parameters; 25 IO hypertensives have developed sustained hypertension. Considering them together with the 19 patients already treated, 72% of 61 IO hypertensives developed ambulatory hypertension over a 3-year period. The patients who subsequently developed hypertension differed from the group who did not only for lower basal values of LV diastolic parameters; all the patients with basal LV hypertrophy and/or preclinical diastolic impairment subsequently developed sustained hypertension. In conclusion, IO hypertensive patients show a high rate of progression towards sustained hypertension. Basal LV hypertrophy and/or preclinical diastolic dysfunction were the only markers of a greater risk of becoming hypertensives. PMID- 16257878 TI - Relationship between custodial status and psychosocial problems among cocaine abusing parents initiating substance abuse treatment. AB - Using the Addiction Severity Index and Brief Symptom Inventory, drug use and psychosocial problems are compared between 93 custodial and 125 non-custodial mothers and fathers initiating outpatient treatment for cocaine dependence. Compared to non-custodial parents, custodial parents experienced more severe current cocaine and alcohol problems, including spending more money on cocaine and alcohol, as well as using more cocaine and being intoxicated on more days. Non-custodial parents demonstrated more psychological distress, more prior history of alcohol problems, and greater current employment and legal problems than custodial parents. Suggestions are made for differential treatment plans based on these findings. PMID- 16257879 TI - From alcoholism treatment to the alcohol harm reduction strategy for England: an overview of alcohol policy since 1950. AB - With the publication of the Alcohol Harm Reduction Strategy for England in 2004,(1) it is timely to reflect on the social and political contexts that have influenced alcohol policy. This paper provides an overview of trends in the development of alcohol policy in England since 1950 with a focus on treatment policy. In particular, it traces factors that have prompted change and resulted in the "treatment" response of the 1960s becoming a small part of a larger, complex approach to the "management" of alcohol-related harm. The publication of the Alcohol Harm Reduction Strategy for England(1) and the Interim Analytical Report,(2) which provided the evidence and framework for the strategy, has resulted in fierce debate on the political processes underlying the emergence of the strategy, the extent to which the strategy is "evidence-based," its strategic aims, and the mechanisms for implementation. This paper argues that responses to policy statements-like the policies themselves-have to be examined within the political, economic, and cultural contexts of their time. PMID- 16257880 TI - Improving continuity of care in a public addiction treatment system with clinical case management. AB - Philadelphia attempted to expand the access to and continuity of addiction treatment by focusing on the 15% of patients who received multiple, detoxification-only (MDO) treatments each year. Clinical Case Managers at five detoxification centers encouraged MDO patients to continue care following detoxification in methadone, residential, or outpatient rehabilitation, and sustain improvements, and they recommended opening detoxification access for additional patients as well. System administrative information was available for one year prior and three years during the intervention. Counts of unduplicated patients within each year and measures of the length and type of treatment episodes determined the intervention effects. Records from a sub-sample of 100 MDO patients were examined to assess specific changes in system utilization. Over three years, 890 MDO patients were case managed and had received assessment, referral, and transport to health care and sober living. The sub-sample of case managed MDO patients showed a 55% reduction in detoxification-only admissions, a 70% increase in use of rehabilitation, and a twenty-day increase in the average length of stay per episode. Though there are noted limitations in the evaluation design, the findings are consistent with the view that individual case management of MDO patients may improve the clinical appropriateness and administrative efficiency of public addiction treatment. PMID- 16257882 TI - Alcohol-related suicides in victims with a history of hospital-treated depression. AB - Victims of suicides are frequently known to have suffered from depression and alcohol-related disorders, but whether these disorders exert different impacts on the period of survival following last hospitalizations have remained unknown. We surveyed 1,585 suicide victims from northern Finland and assigned them to one of three groups, based on lifetime history of depression, alcohol-related disorders, and both together. We then compared survival times in the three groups. Survival times in depressed alcoholic and non-alcoholic males were significantly shorter than those of males with alcohol-related disorders alone. Depressed but non alcoholic suicide victims had more commonly used violent methods, had less often been under the influence of alcohol, and had had more psychotic disorders than the rest. It is apparently important in clinical practice to recognize the increased risk of suicide soon after hospital discharge not only in depressed patients, but also in those with a history of both depression and alcohol-related disorders. PMID- 16257881 TI - Identification and treatment of patients with nicotine problems in routine clinical psychiatry practice. AB - The aim of this study is to assess the rates of nicotine problems diagnosed by psychiatrists, the characteristics of psychiatric patients who smoke, and the services provided to them in routine psychiatric practice. Data were obtained by asking psychiatrists participating in the American Psychiatric Institute for Psychiatric Research and Education's Practice Research Network to complete a self administered questionnaire to provide detailed sociodemographic, clinical, and health plan information on three of their patients seen during routine clinical practice. A total of 615 psychiatrists provided information on 1,843 patients, of which 280 (16.6%) were reported to have a current nicotine problem. Of these, 9.1% were reported to receive treatment for nicotine dependence. Patients with nicotine problems were significantly more likely to be males, divorced or separated, disabled, and uninsured, and have fewer years of education. They also had significantly more co-morbid psychiatric disorders, particularly schizophrenia or alcohol/substance use disorders; a lower Global Assessment Functioning score; and poorer treatment compliance than their counterparts. The results suggest a very low rate of identification and treatment of nicotine problems among patients treated by psychiatrists, even though psychiatric patients who smoke seem to have more clinical and psychosocial stressors and more severe psychiatric problems than those who do not smoke. Programs should be developed to raise the awareness and ability of psychiatrists to diagnose and treat patients with nicotine problems, with a particular emphasis on the increased medical and psychosocial needs of psychiatric patients who smoke. PMID- 16257883 TI - Substance use among school-based youths in northern Mexico. AB - A small base of research suggests that adolescent substance use is a growing public health concern in Mexico. Employing confidential methods, the International Longitudinal Survey of Adolescent Health was administered to assess substance use among 1,238 students in northern Mexico. A large proportion of students indicated lifetime use of tobacco and alcohol. Gender differences in tobacco, alcohol, and marijuana were also evident. The current findings are congruent with the sparse extant data on youths' substance use in Mexico and highlight the need for early prevention interventions. PMID- 16257884 TI - Estimating the prevalence of alcohol and drug abuse and dependence among New Orleans arrestees. AB - The Arrestee Drug Abuse Monitoring Program collects self-report drug use data and urine specimens from adult arrestees across the United States. Despite high levels of drug use among arrestees, few studies have examined need for alcohol and other drug (AOD) treatment within this population. To address this limitation, a secondary analysis was undertaken with a sample of 940 New Orleans arrestees surveyed in 2001. Fifty-nine percent of the sample was diagnosed as needing AOD treatment. Because "need for treatment" was operationalized as a diagnosis of abuse or dependence, the current results should be viewed as liberal estimates of treatment need. Drug treatment implications for criminal populations are discussed. PMID- 16257886 TI - New-onset mania and psychosis following heroin detoxification and naltrexone maintenance. PMID- 16257885 TI - The relationship between future orientation and street substance use among Texas alternative school students. AB - Self-reported substance use data were collected from 963 alternative school students in grades 7-12 who were surveyed through the Safer Choices 2 study in Houston, Texas. Data were collected between October 2000 and March 2001. Logistic regression analyses indicated that lower levels of future orientation was significantly associated (OR = 0.88, 95% CI = 0.81-0.97) with thirty-day substance use after controlling for age and gender. In addition, lower levels of future orientation was found to have a significant association with students' lifetime substance use (OR = 0.93, 95% CI = 0.87-.99) after controlling for age, race, and gender. While the relationships tested in this study are exploratory, they provide evidence for an important connection between future orientation and substance use among adolescents attending alternative schools. PMID- 16257892 TI - [Strategy for the protein identification of human proteome expression profile: selection of searching database]. AB - Widely used method of protein identification for high-throughout proteome expression profile studies was database-dependent, so the selection of databases for the protein identification was very important. Despite the deficiency of available human protein databases, the complementarity of human proteins could be got mainly from human genome but not from the protein databases of other organisms. According to the comparison of the current protein databases from different aspects, IPI was recommended for the basic identification for the studies of human proteome expression profile, and other human protein or nucleic acid databases were needed for the complementary identification and novel protein mining. PMID- 16257893 TI - [The roles of rab5a gene in the mechanism of tumor metastasis]. AB - To study the role of rab5a gene in the mechanism of tumor metastasis, rab5a gene was stable transfected into AGZY83-a cell, a lung adenocarcinoma cell line with low metastatic potential. Superarray was applied to study the effect of rab5a on the differential expression of genes related to tumor metastasis. Five differentially expressed genes were obtained, including one up-regulated gene s100a4 and 4 down-regulated genes, nm23a, rac1, cst3 and col4a2. These results were confirmed at both the RNA and protein levels. We concluded that rab5a promoted tumor metastasis by affecting multiple genes in several pathways involved in tumor metastasis. PMID- 16257894 TI - [Molecular detection of specific HPV types in Condylomata acuminata]. AB - To detect HPV in genital warts (Condylomata acuminata, CA) for infection rate and association of specific HPV types between males and females, and to provide support for the development of HPV vaccines, we designed HPV type-specific oligonucleotide primers to amplify DNA fragments encoding L1 viral capsule protein. SSP-PCR was conducted in duplication for each CA sample from male and female patients. DNA of TA-cloned HPV was used as positive control, and deionized H2O was used as negative control. A total of 22 clinical samples, 13 from males and 9 from females, was collected from patients diagnosed with CA at hospitals in Beijing and Handan. HPV viral DNA was amplified in all 22 samples analyzed, with 100% detection rate. TA-cloning and sequencing of the PCR products confirmed correct amplification of HPV type-specific fragments. Of the 13 samples from males, 5 were infected with HPV6, 6 with HPV11, and 2 with HPV6 + HPV11. Of the 9 samples from females, 3 were infected with HPV6, 2 with HPV11, and 4 with both HPV6 and HPV11 infection. In addition, high-risk types HPV16, HPV18, HPV33, HPV35, HPV45, HPV54, HPV56 and HPV58 were also detected in 4 female samples that were mixed with cervical cell debris during sample collection. However, no HPV types other than HPV6 and HPV11was detected in all CA-only samples in this study. We have established a sensitive and reliable laboratory procedure for HPV detection and classification. Using the method, we reached 100% detection rate of HPV in the CA samples. Our results confirm that HPV6 and HPV11 are primarily responsible for CA, and there is no specific association of HPV types between warts in males and females. PMID- 16257895 TI - [The expression of Staufen gene in colorectal cancer]. AB - To study the expression of differential methylation-associated Staufen gene in colorectal cancer (CRC), we detected the expression of Staufen gene in adenocarcinoma, mucosa adjacent to colorectal cancer (MACC) and corresponding distant normal tissue using RT-PCR and immunohistochemistry. We found that the expression of Staufen gene in MACC and adenocarcinoma of CRC was significantly lower than that in corresponding distal normal tissue at mRNA level (P<0.05.No significant associations were found between age, sex, location of cancer (colon/rectal), degree of differentiation and metastasis of lymph node with expression of Staufen gene. Lower expression of Staufen protein in MACC was verified by comparing with matched normal tissue and adenocarcinoma (P<0.05). This study indicated low expression of Staufen gene in colorectal cancer and the Staufen gene might be involved in the development of colorectal cancer. PMID- 16257896 TI - [The genetic effects of MyoG gene]. AB - The PCR-RFLP technique was applied in this study to analyze the MspI polymorphism in the 3-UTR of MyoG gene. The relationship between different genotypes and corresponding traits and the genetic effects of different allele were analyzed. The results indicated that the N allele has highly significant genetic effects in improving carcass lean percent and the loin eye area, and decreasing the fat content (P<0.01). But no significant influence was found to the FOM carcass traits (P>0.05). As meat quality traits being considered, the N allele highly significantly decreased the pH value, meat color, intramuscular fat content, increased the drip loss (P<0.01) and caused the worse of meat quality. When considering genetic values of different traits, it was found that the N allele had additive effects of 3.929% to carcass lean percent, 2.0985 cm2 to loin eye area, -3.0245% to the fat content, -0.167 to the pH1 value, 0.558% to the drip loss and -0.963% to intramuscular fat content. But no effect was observed to the carcass grading traits. PMID- 16257897 TI - [Phylogenesis analysis of 10 chinese indigenous pig populations]. AB - Genetic relationship of 10 indigenous pig populations was analyzed by 10 microsatellite markers. Results showed that different phylogenetic methods led to different results. The NJ phylogenetic tree from Nei's genetic distance was consistent with the facts. The 10 pig populations were divided into 3 clusters. The first cluster was Guizhou miniature pig and its inbreeding lines, Diannan xiaoer and Wuzhishan miniature pig, the second Yimeng black pig and Rongchang pig, and the third Erhualian, Jinhua and Bama miniature pig. PMID- 16257898 TI - [Sequence variation of TYR exon 1 and origin of pigs]. AB - To investigate the origin and genetic diversity of domestic pigs, the porcine TYR exon 1 in 36 individuals from 12 Chinese indigenous breeds, three European breeds, eight Chinese wild boars and two Vietnamese wild boars was sequenced. Sequence analysis revealed six synonymous mutations, and all the sequences could be sorted into 4 haplotypes. Combining with the published sequences, we constructed a reduced median network (RM network), in which TYR*2 was a haplotype dominated by European domestic pigs and wild boars, plus only three chromosomes from Asian pigs. Most Asian domestic pigs and wild boars shared haplotype TYR*1, demonstrating that TYR*1 was an Asian specific haplotype. Meanwhile, some European domestic pigs and wild boars carried the haplotype TYR*1. TYR*3 and TYR*4 were two haplotypes with low frequencies, containing mainly Chinese indigenous pigs and Asian wild boars, plus some European domestic pigs. Independent domestication of pigs from Asia and Europe was supported by the pattern of RM network. The European commercial breeds had been suffered from introgression from Chinese pigs, and a few Chinese indigenous breeds and Japanese wild boars were also suffered from introgression from European pigs as well. PMID- 16257899 TI - [Analysis of genetic diversity of Sichuan indigenous chicken breeds using microsatellite markers]. AB - Thirty microsatellite markers with medium or high polymorphisms were selected to detect the genetic diversity of 8 indigenous chicken breeds in Sichuan. According to the allele frequencies of 30 microsatellite sites, mean heterozygosity (H), polymorphism information content (PIC) and DA genetic distances were calculated for each breeds. The results showed that 24 of 30 microsatellite sites were highly polymorphic, so the 24 microsatellite markers were effective markers for analysis of genetic relationship among chicken breeds. The mean heterozygosity of 8 chicken breeds was all over 0.5. The highest was the Luning chicken (0.681), and the lowest was the Jiuyuan Dark chicken. The high diversity of 8 chicken breeds might be caused by the traffic obstruction(geographic isolation). The results of the heterozygosity were consistent with that of PIC. UPGMA tree was completed through analysis of DA genetic distances. Emei Dark chicken, Miyi chicken, Luning chicken and Jiuyuan Dark chicken were the first group: Miyi chicken and Luning chicken were grouped firstly, then Emei Dark chicken were grouped with them in shorter time distances, and Jiuyuan Dark chicken were grouped with them at last. Shimiancao Ke chicken Xingwen Silky chicken and Muchuan Silky were the second group: Xingwen Silky chicken and Muchuan Silky were grouped firstly, and then Shimiancao Ke chicken was grouped with them. Liangshangya Ying chicken had its own branch. The result of UPGM was consistent with the genesis, breeding history, differentiation and location of 8 chicken breeds. PMID- 16257900 TI - In vitro expression and antibody preparation of black carp (Mylopharyngodon piceus) GH. AB - The cDNA fragment encoding the mature polypeptide of growth hormone (GH) for the black carp (Mylopharyngodon piceus) was PCR amplified and subcloned into pET-28a. The recombinant expression plasmid pET-bcGH was transformed into E.coli BL21(DE3) and fusion polypeptide containing a 6xHis-tag at the N-terminus was expressed after IPTG induction. The fusion protein band of 23 kDa or so showed immunoreactivity to the polyclonal antibody aganist grass carp GH. The recombinant GH for black carp was purified by affinity chromatography and dialysis. Using the fusion protein as an antigen, through the modified immunization method, the polyclonal antiserum to black carp GH was obtained. Immunochemistry results showed that the antiserum could detect the antigen as low as 4 ng. The protein of 21 kDa in black carp pituitary protein extracts and blood serum could be detected by western blot analysis in which polyclonal antiserum to black carp GH was used as the primary antibody. All these results showed that the polyclonal antiserum against black carp GH was not only effective but also highly specific. PMID- 16257901 TI - [Chromosomal C-banding karyotype of 2 species of genus Asiotmethis (Acridoidea:Pamphagidae) from China]. AB - The chromosomal C-banding karyotype in the spermatogenesis of 2 species of Genus Asiotmethis, Uvarov 1943 from China were investigated. It was found that the diploid chromosome number of Asiotmethis zacharjini (Bei-Bienko, 1926) was 2n (male) =18, the neo-X was a submetacentric chromosome and the rest were acrocentric chromosomes. C-banding pattern showed that all chromosomes presented a paracentromeric block, an interstitial C-band near the centrometic C-block and additional C-band in the distal region. And, the mechanism for sex determination lay in neo-XY (male). In Asiotmethis jubatus (Uvarov, 1926), the diploid chromosome number was 2n (male) =19. All autosomes and X-chromosomes were acrocentric with paracentromeric C-band only, and the mechanism for sex determination lay in XO (male). The difference in heterochromatin content between the 2 species was significant at 0.05 level. PMID- 16257902 TI - [Phylogenetic relationships among domestic goose breeds based on mitochondrial cytochrome b gene sequence variation]. AB - The mitochondrial cytochrome b (cyt b) gene of 17 breeds of domestic goose (2 in Europe and 15 in China) was sequenced and compared with that of the representative of wild species Anser albifrons. Sequence analysis revealed that thirty-one variable sites and 4 different haplotypes were found among the 45 sequences examined, and nucleotide diversity and haplotype diversity were 0.0068 and 0.45, respectively. Mutations of insertion/deletion were not found. Across all sites,the abundance of G was low (14.2%), whereas the percentages of A, T, and C were quite similar among three Anser goose species. The frequency of guanine differs greatly among the three codon positions. Compositional bias is smaller at first and second positions (0.057 and 0.223) than at third positions (0.492), the probability of transition was higher than that of tranversion (Ts/Tv=9.5-19), there was the highest number of transition in the third codon. The phylogenetic analysis support the hypothesis of dual origin of domestic geese. PMID- 16257903 TI - [The discovery of a specific DNA fragment associated with maize cytoplasmic male sterility and its differential display]. AB - Three pairs of PCR primers were designed according to the mitochondrial DNA sequence. PCR amplification was applied to 3 sets of isonuclear alloplasm materials and 3 sets of isoplasm allonuclear materials. Multiplex PCR and general PCR protocol were adopted with total genomic DNA. As for the primers having detected polymorphsim between male sterility and its maintainers, differential display was conducted with mRNA from different development stage of microspore. The results showed as follows: with total genomic DNA template, primer P1-P2 has amplified a specific fragment only in all the male sterile materials, primer P5 P6 has amplified a specific fragment only in maintainer Huangzaosi, primer P3-P4 has no amplification in all the experiment materials. So primer P1-P2 can be used to distinguish male sterile cytoplasm and normal cytoplasm. RT-PCR was conducted with primer P1-P2 in inbred line huangzaosi and 48-2 with male sterile cytoplasm and normal cytoplasm, mRNA was separately isolated from tetrad stage, uninucleate stage and binucleate stage of microspore development, cDNA was obtained with random hexanucleotide primers. With the cDNA template, specific amplified fragments were also detected by primer P1-P2 in the male sterile materials at different development stage of microspore, but there was no amplification by primer P1-P2 in the 2 maintainer lines. This result indicated that primer P1-P2 can be transcripted at 3 development stages of microspore in all male sterile materials, and same transcript was produced by primer P1-P2 among all male sterile materials include 3 sets of isonuclear alloplasm and 3 sets of isoplasm allonuclear. It was suggested from this experiment that the specific DNA sequence detected by primer P1-P2 in all male sterile material total genomic DNA might be related to the cytoplasmic male sterile character. PMID- 16257904 TI - [Genetic diversity of flue-cured tobacco varieties based on ISSR markers]. AB - Genetic diversity was assessed among 24 flue-cured tobacco varieties by ISSR (inter simple sequence repeats). A total of 100 ISSR primers were used to amplify the DNA from these varieties, of which 10 primers produced reproducible amplified products. Using polyacrylamide gel electrophoresis 208 bands were identified, of which 141 bands were polymorphic among the flue-cured tobacco varieties analyzed. Each primer produced 7-37 bands, the length of which ranged 200-2,400 bp. The ratio of polymorphic bands (PPB) was 67.79%. By cluster analysis based on ISSR markers using UPGMA, 24 varieties were divided into 5 major groups, in which the biggest group consisted of 12 varieties derived from Coker319. The genetic similarity index was 0.66-0.85 among 24 flue-cured tobacco varieties. Low genetic diversity among flue-cured tobacco varieties suggested that it is necessary to expand the genetic base of the flue-cured tobacco. 24 varieties could be distinguished by using 2 ISSR markers. The result also indicated that ISSR analysis was suitable for varietal identification and the study on genetic diversity of tobacco germplasm. PMID- 16257905 TI - [Cloning and expression of alpha-D-galactosidase from coffee bean (Coffea liberica & Coffea canephora)]. AB - Alpha-D-galactosidase (alpha-Gal,E.C. 3.2.1.22) is an exo-glycosidase. The enzyme isolated from coffee beans has been well characterized. It has high activity in hydrolyzing the terminal alpha-D-galactoside residues from glycoconjugates on human blood group B erythrocytes, as well as in converting the blood group B into O. A different 1089 bp cDNA open reading frame(ORF) encoding Gal of Coffea liberica & C. canephora was cloned by homology-based RT-PCR. The cloned Gal most closely resembles the corresponding one from C. aribica (98.7% and 99.27% identity). Heterologous overexpression of the two 1.1 kb cDNA fragments was obtained by using one Pichia pastoris stain GS115 and two secret expression vectors, pPICZalphaA and pGAPZalphaA. The expressed protein from P. pastoris stain GS115 was concentrated by ammonium sulfate precipitation and SDS-PAGE assay showed a clear band in the gel. The highest activity of the recombinant enzyme was up to 48.22 U/mL. PMID- 16257906 TI - [RAPD analysis on genetic diversity of the preconcentrated core germplasms of Camellia Sinensis in China]. AB - The study was to evaluate the genetic diversity of 69 tea cultivars of the preconcentrated core germplasms of Camellia Sinensis in China by the random amplified polymorphic DNA (RAPD). Among 50 arbitrary primers, 32 primers could generate enough amplified bands for all the strains in this study. Among a total of 348 bands observed, 328 (94.3%)bands were polymorphic in the 69 cultivars tested except additional 20 cultivars. Genetic distances between the cultivars varied from 0.223 to 0.723. The study indicated that the pre-concentrated core germplasms of Camellia Sinensis in China could well represent the whole collection in respect of genetic structure and genetic diversity and genetic distance. At the same time, it was the best option to establish core collection of Camellia Sinensis in China by combining morphological markers with DNA molecular markers. PMID- 16257907 TI - [Agrobacterium-mediated transformation of rice mature embryos and regeneration of transgenic plants with Metr gene]. AB - The mature embryos of a Japonica rice, Guanglingxiangnuo, were used for the study on optimization of Agrobacterium-mediated transformation. Several factors affecting the transformation were investigated and a suitable transformation system was set up. For this transformation system, the HRM medium, based on the MS medium, was suitable for inducing callus from rice mature embryos. The suitable time span of initial culture in this medium was 7-8 days before co culturing with Agrobacterium and suitable medium for selection was CC medium. Using this transformation system, the Metr gene was introduced into Guanglingxiangnuo, and many transgenic plants were obtained. Most of these transgenic rice plants were confirmed by PCR technique and basta resistance, indicating the T-DNA had been integrated into the genome of transgenic rice plants. PMID- 16257908 TI - [Expression and activity determination of TNFR domain of osteoprotegerin in E.coli and corresponding antibody preparation]. AB - Osteoprotegerin (OPG) plays an important role in the regulation of bone resorption and remodeling. The TNFR domain of OPG, which is involved in the inhibition of formation and activity of osteoclasts, was amplified by PCR and inserted into multiple cloning site of PET-28a. The recombinant plasmid was transferred into E.coli BL21 to express recombinant protein. It was found that expressed product existed in the form of inclusion body. The inclusion body was solubilized, renatured and purified by affinity chromatography. Polyclonal antibodies with high specificity were obtained from the serum of rabbit immunized with purified recombinant protein. Mice were used to determine the hypocalcemic effect of the recombinant protein. Results showed that the recombinant protein expressed in E.coli had the proper bioactivity. PMID- 16257909 TI - [Hairy root induced by wild-type agrobacterium rhizogenes K599 in soybean, cucumber and garden balsam in vivo]. AB - The plantlets of soybean, cucumber and garden balsam were inoculated by wild-type Agrobacterium rhizogenes K599, and hairy root was induced on inoculated sites in vivo. The frequencies of hairy root induction from wound cotyledons of soybean, cucumber and garden balsam were 100%, 65% and 91%, respectively. Moreover, hairy root was induced from healthy cucumber axillary bud with frequency of 10%. PCR analysis of hairy root DNA was conducted using the primers from rolC gene. The PCR results showed that all hairy root lines contained T-DNA. The established system should be ideal for studying soybean and cucumber nematode and garden balsam breeding of flower dwarf architecture. PMID- 16257911 TI - [Gene expression, purification and functional analysis of LiPrmA]. AB - Leptospira interrogans (L. interrogans) genomic DNA was used as template to amplify the full-length gene for ribosomal protein L11 methyltransferase (liPrmA) by PCR. The pET22b-/liprmA expression plasmid was successfully constructed in Escherichia coli (E.coli.) strain TOP10 and confirmed by restriction enzyme digest and sequencing. Through optimizing expression of the recombinant liPrmA 6xHis fusion protein in expression host E. coli. BL21, the yield of soluble target protein reached 40 mg (liter culture)-1. The LiPrmA was purified to apparent homogeneity in a single step using Ni-NTA His Bind chromatography. Amino acid homologous analysis showed that liPrmA shared significant identity with other prokaryotic PrmA and eukaryotic putative PrmA in the catalytic region including AdoMet binding domain. Methylation activity experiments showed purified liPrmA was able to catalyze the ribosomal protein L11 of L. interrogans methylated under the presence of S-adenosyl-methionine (AdoMet). PMID- 16257910 TI - [Molecular diversity of the gene encoding Fe(II)-oxidizing enzyme in Acidithiobacillus ferrooxidans]. AB - Acidithiobacillus ferrooxidans (A.f) is a Gram-negative chemolithotrophic bacterium able to oxidize ferrous iron, elemental sulfur and inorganic sulfur compounds. Fe(II)-oxidizing enzyme plays key role in Fe(II)-oxidizing system. Different A.f strains, which were isolated and purified from various ecological niches, show obviously phenotypic diversity after comparison rates of growth and oxidation of Fe2+. The Fe(II)-oxidizing enzyme gene(iro) for five strains was identified by PCR reactions uitilising primers and genomic DNA, then sequenced and multialignment. The results show there is a putative high variable region in CDS between the 187th and 195th bp and the sequence similarities are from 97% to 99%. For strain YTW, P187-189 Thr-->Pro; For P193-195, Met-->Asn for strain TK; Met-->Ile for strain BY; For P219, all strains T-->C. For upstream of CDS, sequences of all strains are identical to the Genebank sequence (E03451), but downstream of CDS, the sequences exist some variable sites too. PMID- 16257912 TI - [A method for DNA template preparation using a single embryo]. AB - We established a simple method for the preparation of DNA template from a single oocyte or early embryo by KOH/DTT-Triton X disintegration. The PCR amplification efficiency of DNA template prepared by this method was compared with that prepared by TE-proteinase K. Single oocyte, 2-cell embryo, 8-cell embryo, morula or blastocyst were separately treated by KOH/DTT-Triton X, then the DNA template was directly used to amplify mitochondrial DNA segment by PCR. The overall PCR success rate of the 3 pairs of primers was 100% (70/70), while the overall PCR success rate of single oocyte treated by TE-proteinase K was 92.9% (65/70). Difference between the two results was significant (P<0.05), and the PCR false positive rates in both groups were 0. The designed KOH/DTT-Triton X disintegrate method was efficient to the preparation of DNA template of a single early embryo. It needed only one cycle of PCR amplification to get clear aimed DNA stripe and the efficiency was high enough to meet the need of early embryonic genetic material detection. PMID- 16257913 TI - [Application of chromatin immunoprecipitation assay in deciphering DNA-protein interactions]. AB - In the post-genomic era, identifying and characterizing various DNA-protein interactions are a major challenge in the research of gene transcriptional regulation. Although many techniques can be used for this purpose, chromatin immunoprecipitation assay (ChIP), by contrast, is ideally suited for studying DNA protein interactions in vivo. In recent years, standard ChIP assay has been modified to uncover some known factors' unknown target sequences, especially when combined with DNA microarray and molecular cloning strategies. These high throughput ChIP assays are more and more used to reveal the distribution profile of trans-acting factor binding sites throughout the genome, which may yield many new insights into the DNA-protein interaction network. This article summarized the methods of ChIP assay, and highlighted recent progress in the application of this technique. PMID- 16257914 TI - [SSRHunter: development of a local searching software for SSR sites]. AB - Progress in genome research has made it possible to develop new SSR markers by bioinformatics in a relatively narrow region of genome. To realize it, the first thing is to search for potential SSR sites. Any known methods have more or less defects. Efforts were made to develop a local SSR sites searching software in this study. The resultant software, SSRHunter, could accomplish this task perfectly. Furthermore, SSRHunter could provide automatic pretreatment of sequences, routine arrangement of sequences, sequence transformation and convenient report output. PMID- 16257915 TI - [An attempt at reforming genetics experiment for undergraduate course]. AB - The four comprehensive experiments were concreted from a series of verifying genetics experiments, which are relatively specific. They were divided into four pieces of experiment modules: Drosophila melanogaster's classical comprehensive genetics experiment, Bio-Cytogenetics comprehensive experiment, Molecular Genetics comprehensive experiment and Population genetics experiment. The first three are major courses and the last one is a selective course. All of these four require experiment modules. These experiments should be comprehensive, designable and innovative. The lab should open to students the whole day in order to improve students' interest and skill in experiment. The ability to design and innovate experiment could be developed through the training. PMID- 16257916 TI - [The standardization of genetic terms--I. Of the same name with different definition phenomenon]. AB - The phenomenon of same term with different definitions exists widely in genetic teaching courses. The present paper was aiming at analyzing and discussing these problems. For example, the terms such as Y-linked inheritance, sex-limited inheritance, sex-influenced inheritance, haploid, polyploid and cumulative effect expression are very confused and chaotic in meaning, even mutually contradictory. This involves the accuracy of nomenclature. We presumed that the term of Y-linked inheritance cannot confuse with that of sex-limited inheritance, and that though the sex-limited factor and the sex-influenced inheritance gene are both located on autosome with some distance, but they are still different. Haploid refers to the gametophyte chromosome number an individual has, but does not equate to monoploid. Also we thought that polyploid represented the basic number of chromosome, such as triploid indicated for 3x. And, the cumulative effect refers to mutual accumulation function between the exponential quantitative characters of relevant genes. PMID- 16257917 TI - [The relationship between abnormity of synaptonemal complex and male fertility impairment in human]. AB - It was said that 11% of all men showed their infertility. The genetic origins of male infertility may be classified into three main groups: chromosome abnormalities, microdeletions and gene mutations. Growing literature has shown that spermatogenesis failure or reproductive failure (pregnancy wastages) occurred in male carriers of chromosomal distortion/aberration. But the mechanisms remain largely unsolved. Synaptonemal complex (SC) of human spermatocytes from such carriers offers new information. This review aims to summarize recent development of SC analysis for male infertile diagnosis and sum up our results obtained recently. The relationship between male infertility/sterility and SC abnormity was discussed and reviewed as following five aspects. (1) The association of XY-bivalent and the rearranged autosomes interfere with or affect, by their contact, X chromosome normal functions. (2) Male infertility is related to the incomplete pairing in break regions of rearranged autosomes. (3) SC fragmentation, lateral elements (LEs) swelling and pairing disorder result in spermatogenic failure. (4) This heterosynapsis at early stage of meiosis in rearranged autosomes, results in unbalanced germs and pregnancy wastages. (5) Gene mutations of SC proteins result in male infertility. PMID- 16257918 TI - [Advances in the study on molecular mechanism for maintaining the undifferentiated state of embryonic stem cells]. AB - Embryonic stem cells (ES cells), that are derived from the inner cell mass (ICM) of early embryos at the blastocyst stage, have the ability of self-renewal and multiple differentiation capacity. Now ES cells have been extensively applied in basic research and clinical application. That ES cells maintain their undifferentiated state during culture is the prerequisite and basis of their application, and therefore it is necessary to elucidate the molecular mechanism. This review summarized the latest progress in the study on the molecular mechanism for maintaining the undifferentiated state of embryonic stem cells, and emphasized the function of differentiation suppression factors such as LIF, Oct 3/4 and Nanog. PMID- 16257919 TI - [The current feature of the study on human coagulation factor IX mutant]. AB - Hemophilia B is an X-linked bleeding disease, caused by the mutations of human coagulation factor IX (hFIX) gene located in chromosome X. It results in a dramatic decline of hFIX quantity or clotting activity in plasma, and the intrinsic clotting pathway is affected seriously. In this article, the structure and function of hFIX gene as well as the protein encoded by this gene were reviewed. Various types of hFIX mutants found in hemophilia B were also described, including the mutations caused by founder effects, mutations in regulatory region, coding region, splicing sites of introns and two other special mutations. Meanwhile, the biological effects of the mutations described above were discussed. Finally, a mutation type (Arg-->Ala at point 338) that can increase the clotting activity of hFIX as well as the potential application was briefly introduced. PMID- 16257920 TI - [Experimental strategies of the application of RNAi technique in mammalian cells]. AB - As a novel and effective tool for gene function analysis, RNA interference (RNAi) technique has been developed rapidly and made great progress in mammalian cell studies. The mechanisms and experimental operation of RNAi in mammalian cells have become research hotspots, and RNAi mechanisms were supposed to be different from those in plant. In this paper, a review of RNAi mechanisms, the latest research progress and the experimental strategies of RNAi in mammalian cells were presented, including siRNA sequence design, siRNA delivery approaches and RNAi effect detection, so as to offer valuable references for RNAi studies. PMID- 16257921 TI - [Horizontal transfer of environmental pollutant-degrading gene and application in bioremediation]. AB - Horizontal gene transfer, unlike vertical gene transfer, is a means of genetic communication in bacteria. In the special polluted environment, horizontal transfer of polluted-degrading gene has significant functions. Study on horizontal transfer of degrading gene in polluted environment may deepen our understanding of the mechanism of bacterial adaptation to the organic-polluted environment. In the practical application in bioremediation, horizontal transfer of degrading gene can be regulated to promote degrading ability of microorganisms. In this article, we will review the advances in the study on mechanisms of genetic interactions among bacteria, the effect of degrading gene transfer in contaminated environment on microorganisms'adaptation to contaminated environment and the degradation of the pollutants. PMID- 16257922 TI - [Review on hrp genes of plant pathogenic bacteria]. AB - The hrp genes exist in 4 kinds of Gram-negative plant pathogenic bacteria and are responsible for the pathogenicity of bacteria. They can induce hypersensitive response on non-host and resistant plants. In the present paper, we summarized the hrp genes clusters, the relationship between hrp and avr genes, harpin proteins encoded by hrp genes, modulation and function of hrp genes, and plant bacteria interactions mediated by hrp genes in more details. Moreover, trends in future research of plant pathogenic bacteria hrp genes have also been analyzed. PMID- 16257925 TI - Can prenatal N-3 fatty acid deficiency be completely reversed after birth? Effects on retinal and brain biochemistry and visual function in rhesus monkeys. AB - Our previous studies of rhesus monkeys showed that combined prenatal and postnatal n-3 fatty acid deficiency resulted in reduced visual acuity, abnormal retinal function, and low retina and brain docosahexaenoic acid content. We now report effects of n-3 fatty acid deficiency during intrauterine development only. Rhesus infants, born to mothers fed an n-3 fatty acid deficient diet throughout pregnancy, were repleted with a diet high in alpha-linolenic acid from birth to 3 y. Fatty acid composition was determined for plasma and erythrocytes at several time points, for prefrontal cerebral cortex biopsies at 15, 30, 45, and 60 wk, and for cerebral cortex and retina at 3 y. Visual acuity was determined behaviorally at 4, 8, and 12 postnatal weeks, and the electroretinogram was recorded at 3-4 mo. Total n-3 fatty acids were reduced by 70-90% in plasma, erythrocytes, and tissues at birth but recovered to control values within 4 wk in plasma, 8 wk in erythrocytes, and 15 wk in cerebral cortex. At 3 y, fatty acid composition was normal in brain phospholipids, but in the retina DHA recovery was incomplete (84% of controls). Visual acuity thresholds did not differ from those of control infants from mothers fed a high linolenic acid diet. However, the repleted group had lower amplitudes of cone and rod ERG a-waves. These data suggest that restriction of n-3 fatty acid intake during the prenatal period may have long-term effects on retinal fatty acid composition and function. PMID- 16257923 TI - Differential induction of PPAR-gamma by luminal glutamine and iNOS by luminal arginine in the rodent postischemic small bowel. AB - Using a rodent model of gut ischemia-reperfusion (I/R), we have previously shown that the induction of inducible nitric oxide synthase (iNOS) is harmful, whereas the induction of heme oxygenase 1 (HO-1) and peroxisome proliferator-activated receptor-gamma (PPAR-gamma) is protective. In the present study, we hypothesized that the luminal nutrients arginine and glutamine differentially modulate these molecular events in the postischemic gut. Jejunal sacs were created in rats at laparotomy, filled with either 60 mM glutamine, arginine, or magnesium sulfate (osmotic control) followed by 60 min of superior mesenteric artery occlusion and 6 h of reperfusion, and compared with shams. The jejunum was harvested for histology or myeloperoxidase (MPO) activity (inflammation). Heat shock proteins and iNOS were quantitated by Western blot analysis and PPAR-gamma by DNA binding activity. In some experiments, rats were pretreated with the PPAR-gamma inhibitor G9662 or with the iNOS inhibitor N-[3(aminomethyl)benzyl]acetamidine (1400W). iNOS was significantly increased by arginine but not by glutamine following gut I/R and was associated with increased MPO activity and mucosal injury. On the other hand, PPAR-gamma was significantly increased by glutamine but decreased by arginine, whereas heat shock proteins were similarly increased in all experimental groups. The PPAR-gamma inhibitor G9662 abrogated the protective effects of glutamine, whereas the iNOS inhibitor 1400W attenuated the injurious effects of arginine. We concluded that luminal arginine and glutamine differentially modulate the molecular events that regulate injurious I/R-mediated gut inflammation and injury. The induction of PPAR-gamma by luminal glutamine is a novel protective mechanism, whereas luminal arginine appears harmful to the postischemic gut due to enhanced expression of iNOS. PMID- 16257926 TI - The role of UGT1A1*28 mutation in jaundiced infants with hypertrophic pyloric stenosis. AB - Hypertrophic pyloric stenosis (HPS) may be accompanied by jaundice, a condition referred to as the icteropyloric syndrome (IPS). It has long been suspected that the etiology of IPS is an early manifestation of Gilbert's syndrome (GS). Clinical features common to both GS and IPS include jaundice precipitated by fasting and improved with feeding. Prevalence of jaundice in HPS is similar to that of clinically apparent GS in the general population. Discovery of a mutation in the promoter region of the bilirubin uridine diphosphate glucuronosyl transferase gene (UGT1A1*28) as the most common cause of GS has provided a tool to determine the role of GS in IPS. The aims of this study were to determine 1) the prevalence of IPS in a large group of infants with HPS, 2) whether disease severity contributed to the manifestation of IPS, and 3) whether GS played a role in IPS. Radioactive PCR and sequencing were used to determine the presence of UGT1A1*28 mutations. We determined a prevalence of IPS of 14.3% in HPS. Infants with IPS had significantly higher levels of alkalosis than infants with HPS alone. GS mutations were 4-fold higher in IPS (43.8%) than HPS (10.7%). In conclusion, the frequency of jaundice in HPS is similar to that of clinically apparent GS in the general population. Manifestation of IPS results from a more severe degree of metabolic disturbance and the presence of GS mutations. PMID- 16257927 TI - Developmental changes in intraepithelial T lymphocytes and NK cells in the small intestine of neonatal rats. AB - The main objective of this study was to characterize developmental changes in small intestinal intraepithelial lymphocyte (IEL) subpopulations during the suckling period, thus contributing to the understanding of the development of diffuse gut-associated lymphoid tissue (GALT) and to the identification of early mechanisms that protect the neonate from the first contact with diet and gut microbial antigens. The study was performed by double labeling and flow cytometry in IEL isolated from the proximal and distal small intestine of 1- to 21-d-old Lewis rats. During the suckling period, intraepithelial natural killer (NK) cells changed from a typical systemic phenotype, CD8+, to a specific intestinal phenotype, CD8-. Analysis of CD8+ IEL revealed a progressive increase in the relative number of CD8+ IEL co-expressing TCRalphabeta, cells associated with acquired immunity, whereas the percentage of CD8+ cells expressing the NK receptor, i.e. cells committed to innate immunity, decreased. At weaning, IEL maturity was still not achieved, as revealed by a phenotypic pattern that differed from that of adult rats. Thus, late after weaning, the regulatory CD8+CD4+ T IEL population appeared and the NK population declined. In summary, the intestinal intraepithelial compartment undergoes changes in its lymphocyte composition associated with the first ingestion of food. These changes are focused on a relatively high proportion of NK cells during the suckling period, and after weaning, an expansion of the regulatory CD8+CD4+ T cells. PMID- 16257928 TI - The effects of hypercapnia on cerebral autoregulation in ventilated very low birth weight infants. AB - Permissive hypercapnia, a strategy allowing high Pa(CO2), is widely used by neonatologists to minimize lung damage in ventilated very low birth weight (VLBW) infants. While hypercapnia increases cerebral blood flow (CBF), its effects on cerebral autoregulation of VLBW infants are unknown. Monitoring of mean CBF velocity (mCBFv), Pa(CO2), and mean arterial blood pressure (MABP) from 43 ventilated VLBW infants during the first week of life was performed during and after 117 tracheal suctioning procedures. Autoregulation status was determined during tracheal suctioning because it perturbs cerebral and systemic hemodynamics. The slope of the relationship between mCBFv and MABP was estimated when Pa(CO2) was fixed at 30, 35, 40, 45, 50, 55, and 60 mm Hg. A slope near or equal to 0 suggests intact autoregulation, i.e. CBF is not influenced by MABP. Increasing values >0 indicate progressively impaired autoregulation. Infants weighed 905 +/- 259 g and were 26.9 +/- 2.3 wk gestation. The autoregulatory slope increased as Pa(CO2)) increased from 30 to 60 mm Hg. While the slopes for Pa(CO2) values of 30 to 40 mm Hg were not statistically different from 0, slopes for Pa(CO2) > or = 45 mm Hg indicated a progressive loss of cerebral autoregulation. The autoregulatory slope increases with increasing Pa(CO2), suggesting the cerebral circulation becomes progressively pressure passive with hypercapnia. These data raise concerns regarding the use of permissive hypercapnia in ventilated VLBW infants during the first week of life, as impaired autoregulation during this period may be associated with increased vulnerability to brain injury. PMID- 16257929 TI - Diffusion tensor imaging on teenagers, born at term with moderate hypoxic ischemic encephalopathy. AB - Hypoxic-ischemic encephalopathy (HIE) is graded with three levels of severity mild, moderate and severe. The outcome of individuals with mild and severe grades can be reliably predicted from this scheme. Individuals with moderate degree are divided in outcome between those who suffer major neurologic problems (e.g., cerebral palsy) and those who are assumed to recover from the incident. It is however not clear if the recovery is complete and unquestionable. A group of adolescents who had been born at term, diagnosed with moderate HIE but had not developed cerebral palsy, were investigated with diffusion tensor imaging. Fractional anisotropy maps were used as a basis of comparison to a group of controls of the same age and gender distribution. In several white matter areas fractional anisotrophy was lower in the group of individuals with a history of moderate HIE. These areas include the internal capsules (bilaterally in the posterior limb and on the right in the anterior limb), the posterior and anterior corpus callosum as well as frontal inferior white matter areas. These results indicate that even in the absence of such major neurologic impairments as cerebral palsy, moderate HIE causes long term white matter disturbances which are not repaired by adolescence. PMID- 16257930 TI - In vivo constriction of the fetal and neonatal ductus arteriosus by a prostanoid EP4-receptor antagonist in rats. AB - Indomethacin is used to constrict the patent ductus arteriosus in premature infants. To clarify possible prostanoid receptor antagonists that can constrict the ductus, we studied in vivo constriction of the fetal and neonatal ductus arteriosus by AE3-208, a prostanoid EP4-receptor antagonist, in rats. Following quick cesarean section of near-term pregnant rats (21 d), neonates were incubated in room air at 33 degrees C. The inner diameter of the ductus was measured with a microscope and a micrometer following rapid whole-body freezing of the fetus and neonate, and sectioning of the thorax in the frontal plane on a freezing microtome. In the control, the ductus arteriosus constricted quickly after birth, and the inner diameter was 0.80 mm in the fetus and 0.06 mm at 90 min after birth. AE3-208, administered orogastrically to the dam, constricted the fetal ductus dose dependently. Maximal ductal constriction was observed 4 h after administration, and the ductal diameters were 0.06 mm and 0.26 mm after administration of 10 mg/kg and 10 ng/kg of AE3-208, respectively. In neonatal rats, AE3-208 injected subcutaneously at 30 min after birth, inhibited dilatation of the ductus by PGE1 dose dependently. PGE1 (10 microg/kg) was injected subcutaneously to the 1-h-old neonatal rat, and the ductal diameters were 0.53 mm and 0.19 mm without and with pretreatment of AE3-208 (10 microg/kg), respectively. These results indicate the major role of EP4 in the fetal and neonatal ductus and show that an EP4 antagonist can be used to constrict the patent ductus of premature infants. PMID- 16257932 TI - Disparities in global tobacco harm reduction. PMID- 16257934 TI - Challenges and opportunities for ethics in public health. PMID- 16257933 TI - The role of race and genetics in health disparities research. PMID- 16257935 TI - "What would Durkheim do?" a comment on Kushner and Sterk. PMID- 16257936 TI - Trading tobacco: are youths choosing cigars over cigarettes? PMID- 16257937 TI - Stress, trauma, and coronary heart disease among Native Americans. PMID- 16257938 TI - Skin color, social classification, and blood pressure in southeastern Puerto Rico. AB - OBJECTIVES: We tested competing hypotheses for the skin color-blood pressure relationship by analyzing the association between blood pressure and 2 skin color variables: skin pigmentation and social classification. METHODS: We measured skin pigmentation by reflectance spectrophotometry and social classification by linking respondents to ethnographic data on the cultural model of "color" in southeastern Puerto Rico. We used multiple regression analysis to test the associations between these variables and blood pressure in a community-based sample of Puerto Rican adults aged 25-55 years (n=100). Regression models included age, gender, body mass index (BMI), self-reported use of antihypertensive medication, and socioeconomic status (SES). RESULTS: Social classification, but not skin pigmentation, is associated with systolic and diastolic blood pressure through a statistical interaction with SES, independent of age, gender, BMI, self-reported use of antihypertensive medication, and skin reflectance. CONCLUSION: Our findings suggest that sociocultural processes mediate the relationship between skin color and blood pressure. They also help to clarify the meaning and measurement of skin color and "race" as social variables in health research. PMID- 16257939 TI - Racializing drug design: implications of pharmacogenomics for health disparities. AB - Current practices of using "race" in pharmacogenomics research demands consideration of the ethical and social implications for understandings of group difference and for efforts to eliminate health disparities. This discussion focuses on an "infrastructure of racialization" created by current trajectories of research on genetic differences among racially identified groups, the use of race as a proxy for risk in clinical practice, and increasing interest in new market niches by the pharmaceutical industry. The confluence of these factors has resulted in the conflation of genes, disease, and race. I argue that public investment in pharmacogenomics requires careful consideration of current inequities in health status and social and ethical concerns over reifying race and issues of distributive justice. PMID- 16257941 TI - Stormy weather: race, gene expression, and the science of health disparities. AB - In the current US political climate, conservative foundations are seeking to frame debates over determinants of racial/ethnic health disparities as a matter of "politically correct" unscientific ideology (concerning the health impacts of discrimination) vs scientific yet "politically incorrect" expertise rooted in biological facts (concerning genes). I draw on historical and contemporary examples to place conservative polemics in context, and also highlight fundamental flaws in their arguments involving the use of spurious categories (e.g., Caucasian), logical fallacies, temporal fallacies, and an erroneous emphasis on gene frequency over gene expression. The larger goal is to strengthen development of a more critical, reflexive, and rigorous science capable of generating evidence useful for rectifying--rather than perpetuating--social disparities in health. PMID- 16257940 TI - Latino populations: a unique opportunity for the study of race, genetics, and social environment in epidemiological research. AB - Latinos are the largest minority population in the United States. Although usually classified as a single ethnic group by researchers, Latinos are heterogeneous from cultural, socioeconomic, and genetic perspectives. From a cultural and social perspective, Latinos represent a wide variety of national origins and ethnic and cultural groups, with a full spectrum of social class. From a genetic perspective, Latinos are descended from indigenous American, European, and African populations. We review the historical events that led to the formation of contemporary Latino populations and use results from recent genetic and clinical studies to illustrate the unique opportunity Latino groups offer for studying the interaction between racial, genetic, and environmental contributions to disease occurrence and drug response. PMID- 16257942 TI - Genomics and the public health code of ethics. AB - We consider the public health applications of genomic technologies as viewed through the lens of the public health code of ethics. We note, for example, the potential for genomics to increase our appreciation for the public health value of interdependence, the potential for some genomic tools to exacerbate health disparities because of their inaccessibility by the poor and the way in which genomics forces public health to refine its notions of prevention. The public health code of ethics sheds light on concerns raised by commercial genomic products that are not discussed in detail by more clinically oriented perspectives. In addition, the concerns raised by genomics highlight areas of our understanding of the ethical principles of public health in which further refinement may be necessary. PMID- 16257943 TI - Changes in veterans' use of outpatient care from 1992 to 2000. AB - OBJECTIVES: During the mid-1990s, the Veterans Health Administration (VHA) reorganized and placed greater emphasis on high-quality primary care. To determine whether the reorganization was associated with changes in patterns of out-patient VHA use, we sought to evaluate changes in characteristics of veterans who use VHA outpatient services between 1992 and 2000. METHODS: We merged 2 waves of the National Survey of Veterans to determine changes in patterns of outpatient care use. We evaluated the extent to which veterans who received outpatient care received that care from the VHA. RESULTS: The odds ratio for VHA-only outpatient care relative to non-VHA-only care in 2000 relative to 1992 was 1.75 (95% confidence interval [CI]=1.51, 2.04), and the odds ratio for dual relative to non VHA-only care was 1.22 (95% CI=1.08, 1.37). Veterans who were older, had low incomes, and had no additional health insurance coverage were most likely to increase their use of VHA outpatient care. CONCLUSIONS: Our results suggest that the VHA is increasingly serving veterans who have trouble accessing the private health care system. PMID- 16257944 TI - Social disparities in maternal morbidity during labor and delivery between Mexican-born and US-born White Californians, 1996-1998. AB - OBJECTIVES: To assess maternal health disparities, we compared maternal morbidities during labor and delivery among Mexican-born and US-born White, non Latina women residing in California. METHODS: This population-based study used linked hospital discharge and birth certificate data for 1996-1998 (862,723 deliveries). We calculated the frequency, and observed and adjusted odds ratios for obstetric complications. Covariates included maternal age, parity, education, prenatal care initiation and payment source, and hospital quality of care. RESULTS: Approximately 1 in 5 deliveries resulted in a obstetric complication. After control for covariates, Mexican-born women were significantly less likely to have 1 or more maternal morbidities than White, non-Latina women but more likely to have complications that reflect the quality of intrapartum care. CONCLUSIONS: Maternal morbidities during labor and delivery are a substantial burden for women in California. The favorable overall outcome of Mexican-born women over US-born White, non-Latinas is surprising given their lower educational attainment, relative poverty, and greater barriers to health care access. The favorable outcomes obscure vulnerabilities in those complications that are sensitive to the quality of intrapartum care. PMID- 16257945 TI - Breast cancer treatment experiences of Latinas in Los Angeles County. AB - OBJECTIVE: We examined breast cancer treatment experiences of and outcomes for Latinas in Los Angeles County. METHODS: We conducted a population-based survey of women who were diagnosed with breast cancer between December 2001 and November 2002 (n=910) to evaluate the types of treatments received, communication with clinicians, and satisfaction. RESULTS: About two thirds were non-Latina White, 18.8% were African American, and 18.9% were Latina (with 11.0% preferring English and 7.9% preferring Spanish). The rest indicated other ethnic groups. Latinas who preferred Spanish were more likely to experience a delay of 3 months or more from diagnosis to surgical treatment (36.4% vs 9.1% for non-Latina Whites, 18.6% for African Americans, and 12.7%, for other Latinas, P<.001). African Americans and Latinas who preferred Spanish had very low rates of reconstruction (13.8% and 9.2%, respectively, compared with 42.1% for Whites and 34.5% for Latinas who preferred English, P=.009). Latinas who preferred Spanish had the highest odds ratio for low satisfaction. CONCLUSION: Latinas who preferred Spanish received different treatments and perceived a different treatment experience than did other cultural groups. PMID- 16257946 TI - Cancer disparities research partnership in Lakota Country: clinical trials, patient services, and community education for the Oglala, Rosebud, and Cheyenne River Sioux tribes. AB - Native Americans served by the Aberdeen, Billings, and Bemidji areas of the Indian Health Service (IHS) have a cancer mortality rate approximately 40% higher than that of the overall US population. The National Cancer Institute has funded Rapid City Regional Hospital to provide clinical trials, behavioral research, a genetic protocol, patient navigator services (assisting patients with health care coordination and financial issues and helping them to understand their options), and community education for members of 3 western South Dakota tribes. Challenges faced by the project included obtaining multiple approvals from 3 tribes, 4 IHS facilities, and 5 institutional review boards; travel distances; lack of screening; red tape of referrals; and refusal by some payers to cover clinical trials. Building trust through ongoing communication and community presence is key to a successful project. PMID- 16257947 TI - Information technology as a tool to improve the quality of American Indian health care. AB - The American Indian/Alaska Native population experiences a disproportionate burden of disease across a spectrum of conditions. While the recent National Healthcare Disparities Report highlighted differences in quality of care among racial and ethnic groups, there was only very limited information available for American Indians. The Indian Health Service (IHS) is currently enhancing its information systems to improve the measurement of health care quality as well as to support quality improvement initiatives. We summarize current knowledge regarding health care quality for American Indians, highlighting the variation in reported measures in the existing literature. We then discuss how the IHS is using information systems to produce standardized performance measures and present future directions for improving American Indian health care quality. PMID- 16257948 TI - Rural American Indian Medicaid health care services use and health care costs in California. AB - OBJECTIVES: We determined differences in Medicaid service use and health care costs in a rural Indian Health Service (IHS) user population of American Indians and Alaska Natives as compared with Whites. METHODS: California Medicaid eligibility and claims files were linked to IHS user files to obtain a sample of Medicaid-eligible American Indian/Alaska Native users (n=7910). A random sample of Whites was matched for age, gender, aid category, length of eligibility, and county of residence (n=15075). We used generalized linear models to compare risk adjusted use of resources-ambulatory visits, prescriptions, emergency room visits, hospitalizations, and costs-both adjusting and stratifying for dominant source of ambulatory visits. RESULTS: American Indians/Alaska Natives had significantly lower use of Medicaid-paid ambulatory visits, prescriptions, emergency room visits, and hospitalizations and lower associated costs than Whites. Medicaid-paid total costs and use of services were lower for those who predominantly used Indian health program clinics, as well as for those who predominantly used other sources of ambulatory care. CONCLUSIONS: Barriers to receiving Medicaid services and payments exist for American Indians/Alaska Natives in the rural IHS-user population. If American Indians/Alaska Natives are to have Medicaid resources comparable to those of Whites, these barriers must be reduced. PMID- 16257949 TI - Physician-patient racial concordance, continuity of care, and patterns of care for hypertension. AB - To assess the effects of physician-patient racial concordance and continuity of care on hypertension outcomes, we described patterns of care for hypertension; we used cross-tabulations and repeated measures (generalized estimating equations) analyses with panel survey data from elderly persons interviewed and examined in 1987 and 1990. Continuity of care was associated with recognition of hypertension, receipt of medication, and lower incidence of undetected hypertension. Physician race had little effect, but continuity is important for successful management of hypertension in older persons. PMID- 16257950 TI - Building geographic information system capacity in local health departments: lessons from a North Carolina project. AB - State government, university, and local health department (LHD) partners collaborated to build the geographic information system (GIS) capacity of 5 LHDs in North Carolina. Project elements included procuring hardware and software, conducting individualized and group training, developing data layers, guiding the project development process, coordinating participation in technical conferences, providing ongoing project consultation, and evaluating project milestones. The project provided health department personnel with the skills and resources required to use sophisticated information management systems, particularly those that address spatial dimensions of public health practice. This capacity-building project helped LHDs incorporate GIS technology into daily operations, resulting in improved time and cost efficiency. Keys to success included (1) methods training rooted in problems specific to the LHD, (2) required project identification by LHD staff with associated timelines for development, (3) ongoing technical support as staff returned to home offices after training, (4) subgrants to LHDs to ease hardware and software resource constraints, (5) networks of relationships among LHDs and other professional GIS users, and (6) senior LHD leadership who supported the professional development activities being undertaken by staff. PMID- 16257951 TI - Racial/ethnic variations in veterans' ambulatory care use. AB - OBJECTIVES: We assessed racial/ethnic variations in patterns of ambulatory care use among Department of Veterans Affairs (VA) health care-eligible veterans to determine if racial/ethnic differences in health care use persist in equal-access systems. METHODS: We surveyed 3227 male veterans about their health and ambulatory care use. RESULTS: Thirty-eight percent of respondents had not had a health care visit in the previous 12 months. Black (odds ratio [OR] = 0.5), Hispanic (OR = 0.4), and Asian/Pacific Islander veterans (OR=0.4) were less likely than White veterans to report any ambulatory care use. Alternately, Whites (OR=2.2) were more likely than other groups to report ambulatory care use. Being White was a greater predictor of health care use than was having fair or poor health (OR=1.4) or functional limitations (OR=1.5). In non-VA settings, racial/ethnic minorities were less likely to have a usual provider of health care. There was no VA racial/ethnic variation in this parameter. CONCLUSIONS: Racial/ethnic disparities in health and health care use are present among VA health care-eligible veterans. Although the VA plays an important role in health care delivery to ethnic minority veterans, barriers to VA ambulatory care use and additional facilitators for reducing unmet need still need to be investigated. PMID- 16257952 TI - The changing fate of pneumonia as a public health concern in 20th-century America and beyond. AB - For a brief period from the 1930s through the early 1940s, public health advocates made pneumonia a leading public health concern. Predicated on the need for antipneumococcal antiserum, but also incorporating physician reeducation, state "pneumonia control programs" were established nationwide. However, with the advent of penicillin and the sulfonamides, the pneumonia control programs soon collapsed. Pneumonia reverted to the domain of the private practitioner, which was devoid of state oversight. With the emergence of pneumococcal antibiotic resistance in the 1990s, the possibility again arose that pneumonia could become a public health concern, given the nationwide need to curb unnecessary antibiotic usage and to encourage vaccination. An understanding of the history of pneumonia's changing status could shed light on current attempts to reformulate the disease and elucidate the contested domains of private practice and public health. PMID- 16257953 TI - Analysis of NOD2-mediated proteome response to muramyl dipeptide in HEK293 cells. AB - NOD2, a cytosolic receptor for the bacterial proteoglycan fragment muramyl dipeptide (MDP), plays an important role in the recognition of intracellular pathogens. Variants in the bacterial sensor domain of NOD2 are genetically associated with an increased risk for the development of Crohn disease, a human chronic inflammatory bowel disease. In the present study, global protein expression changes after MDP stimulation were analyzed by two-dimensional PAGE of total protein extracts of human cultured cells stably transfected with expression constructs encoding for wild type NOD2 (NOD2(WT)) or the disease-associated NOD2 L1007fsinsC (NOD2(SNP13)) variant. Differentially regulated proteins were identified by matrix-assisted laser desorption ionization time-of-flight (MALDI TOF) mass spectrometry (MS) peptide mass fingerprinting and MALDI MS/MS. The limited overlap in the responses of the NOD2-overexpressing cell lines to MDP included a down-regulation of heat shock 70-kDa protein 4. A complex pro inflammatory program regulated by NOD2(WT) that encompasses a regulation of key genes involved in protein folding, DNA repair, cellular redox homeostasis, and metabolism was observed both under normal growth conditions and after stimulation with MDP. By using the comparison of NOD2(WT) and disease-associated NOD2(SNP13) variant, we have identified a proteomic signature pattern that may further our understanding of the influence of genetic variations in the NOD2 gene in the pathophysiology of chronic inflammatory bowel disease. PMID- 16257954 TI - Cell wall-targeting domain of glycylglycine endopeptidase distinguishes among peptidoglycan cross-bridges. AB - ALE-1, a homologue of lysostaphin, is a peptidoglycan hydrolase that specifically lyses Staphylococcus aureus cell walls by cleaving the pentaglycine linkage between the peptidoglycan chains. Binding of ALE-1 to S. aureus cells through its C-terminal 92 residues, known as the targeting domain, is functionally important for staphylolytic activity. The ALE-1-targeting domain belongs to the SH3b domain family, the prokaryotic counterpart of the eukaryotic SH3 domains. The 1.75 angstroms crystal structure of the targeting domain shows an all-beta fold similar to typical SH3s but with unique features. The structure reveals patches of conserved residues among orthologous targeting domains, forming surface regions that can potentially interact with some common features of the Gram positive cell wall. ALE-1-targeting domain binding studies employing various bacterial peptidoglycans demonstrate that the length of the interpeptide bridge, as well as the amino acid composition of the peptide, confers the maximum binding of the targeting domain to the staphylococcal peptidoglycan. Truncation of the highly conserved first 9 N-terminal residues results in loss of specificity to S. aureus cell wall-targeting, suggesting that these residues confer specificity to S. aureus cell wall. PMID- 16257956 TI - Differential inhibition of mRNA degradation pathways by novel cap analogs. AB - mRNA degradation predominantly proceeds through two alternative routes: the 5'- >3' pathway, which requires deadenylation followed by decapping and 5'-->3' hydrolysis; and the 3'-->5' pathway, which involves deadenylation followed by 3'- >5' hydrolysis and finally decapping. The mechanisms and relative contributions of each pathway are not fully understood. We investigated the effects of different cap structure (Gp(3)G, m(7)Gp(3)G, or m(2)(7,3'-O) Gp(3)G) and 3' termini (A(31),A(60), or G(16)) on both translation and mRNA degradation in mammalian cells. The results indicated that cap structures that bind eIF4E with higher affinity stabilize mRNA to degradation in vivo. mRNA stability depends on the ability of the 5' terminus to bind eIF4E, not merely the presence of a blocking group at the 5'-end. Introducing a stem-loop in the 5'-UTR that dramatically reduces translation, but keeping the cap structure the same, does not alter the rate of mRNA degradation. To test the relative contributions of the 5'-->3' versus 3'-->5' pathways, we designed and synthesized two new cap analogs, in which a methylene group was substituted between the alpha- and beta-phosphate moieties, m(2)(7,3'-O)Gpp(CH2)pG and m(2)(7,3'-O)Gp(CH2)ppG, that are predicted to be resistant to cleavage by Dcp1/Dcp2 and DcpS, respectively. These cap analogs were recognized by eIF4E and conferred cap-dependent translation to mRNA both in vitro and in vivo. Oligonucleotides capped with m(2)(7,3'-O)Gpp(CH2)pG were resistant to hydrolysis by recombinant human Dcp2 in vitro. mRNAs capped with m(2)(7,3'-O)Gpp(CH2)pG, but not m(2)(7,3'-O)Gp(CH2)ppG, were more stable in vivo, indicating that the 5'-->3' pathway makes a major contribution to overall degradation. Luciferase mRNA containing a 5'-terminal m(2)(7,3'-O)Gpp(CH2)pG and 3'-terminal poly(G) had the greatest stability of all mRNAs tested. PMID- 16257955 TI - Versican/PG-M regulates chondrogenesis as an extracellular matrix molecule crucial for mesenchymal condensation. AB - Mesenchymal cell condensation is an essential step for cartilage development. Versican/PG-M, a large chondroitin sulfate proteoglycan, is one of the major molecules expressed in the extracellular matrix during condensation. However, its role, especially as an environment for cells being condensed, has not been elucidated. Here we showed several lines of evidence for essential roles of versican/PG-M in chondrogenic condensation using a new chondrocytic cell line, N1511. Chondrogenic stimuli (treatment with parathyroid hormone, dexamethasone, 10% serum) induced a marked increase in the transcription and protein synthesis of versican/PG-M. Stable antisense clones for versican/PG-M, depending on suppression of the expression of versican/PG-M, showed different capacities for chondrogenesis, as indicated by the expression and deposition of aggrecan, a major chondrocytic cell product. The cells in the early stages of the culture only expressed V0 and V1 forms, having more chondroitin sulfate chains among the four variants of versican/PG-M, and treatment of those cells with chondroitinase ABC suppressed subsequent chondrogenesis. Furthermore, treatment with beta xyloside, an artificial chain initiator of chondroitin sulfate synthesis to consequently inhibit the synthesis on the core proteins, suppressed chondrogenesis. In addition, forced expression of the variant V3, which has no chondroitin sulfate chain, disrupted the deposition and organization of native versican/PG-M (V0/V1) and other extracellular matrix molecules known to be expressed during the mesenchymal condensation and resulted in the inhibition of subsequent chondrogenesis. These results suggest that versican/PG-M is involved in positively regulating the formation of the mesenchymal matrix and the onset of chondrocyte differentiation through the attached chondroitin sulfate chains. PMID- 16257957 TI - Regulation of transactivation function of the aryl hydrocarbon receptor by the Epstein-Barr virus-encoded EBNA-3 protein. AB - EBNA-3 is one of the Epstein-Barr virus (EBV)-encoded nuclear antigens that is indispensable for immunoblastic transformation and sustained proliferation of B lymphocytes. The molecular mechanisms responsible for the function of EBNA-3 are poorly understood. We previously found that EBNA-3 interacts with an immunophilin like protein XAP2/ARA9/AIP, which in mammalian cells is known to interact with the latent aryl hydrocarbon receptor (AhR). AhR is a ligand-inducible transcription factor that mediates cellular responses to environmental pollutants, such as 2,3,7,8-tetrachloro-dibenzo-p-dioxin (TCDD). In this study, we show that EBNA-3 interacts specifically with AhR. The stability of this interaction is determined by the activation state of AhR and its association with XAP2. We and others have demonstrated that XAP2 retains the nonactivated AhR in the cell cytoplasm. However, in the presence of TCDD, the effect of XAP2 on the intracellular localization of AhR was counter-acted by EBNA-3, resulting in nuclear translocation of the AhR. In addition, EBNA-3 enhanced transactivation function by the ligand-activated AhR in cells, as assessed by reporter gene assays. Our data suggested that EBNA-3 plays a role in facilitating the ligand dependent AhR activation process. Following activation of the AhR, we also observed that EBNA-3 counteracted the inhibitory effect of TCDD on the growth of EBV-carrying lymphoblasts. Taken together, our studies revealed a novel interaction between EBV- and AhR-dependent cellular pathways that control cell proliferation and survival. PMID- 16257958 TI - A role for Kif17 in transport of Kv4.2. AB - Although kinesins are known to transport neuronal proteins, it is not known what role they play in the targeting of their cargos to specific subcellular compartments in neurons. Here we present evidence that the K+ channel Kv4.2, which is a major regulator of dendritic excitability, is transported to dendrites by the kinesin isoform Kif17. We show that a dominant negative construct against Kif17 dramatically inhibits localization to dendrites of both introduced and endogenous Kv4.2, but those against other kinesins found in dendrites do not. Kv4.2 colocalizes with Kif17 but not with other kinesin isoforms in dendrites of cortical neurons. Native Kv4.2 and Kif17 coimmunoprecipitate from brain lysate, and introduced, tagged versions of the two proteins coimmunoprecipitate from COS cell lysate, indicating that the two proteins interact, either directly or indirectly. The interaction between Kif17 and Kv4.2 appears to occur through the extreme C terminus of Kv4.2 and not through the dileucine motif. Thus, the dileucine motif does not determine the localization of Kv4.2 by causing the channel to interact with a specific motor protein. In support of this conclusion, we found that the dileucine motif mediates dendritic targeting of CD8 independent of Kif17. Together our data show that Kif17 is probably the motor that transports Kv4.2 to dendrites but suggest that this motor does not, by itself, specify dendritic localization of the channel. PMID- 16257960 TI - LosA, a key glycosyltransferase involved in the biosynthesis of a novel family of glycosylated acyltrehalose lipooligosaccharides from Mycobacterium marinum. AB - Members of the genus Mycobacterium are characterized by cell envelopes rich in unusual free lipids, interacting with a covalently anchored mycolyl arabinogalactan matrix. Previous studies have shown that Mycobacterium marinum produces large amounts of a diacylglycosylphenolphthiocerol, "phenolic" glycolipid. When cultivated on liquid Sauton medium, traces of a polar lipooligosaccharide (LOS) glycolipid antigen were also previously indicated. In this study, it was found that growth of the type strain of M. marinum on solid Sauton or Middlebrook 7H10 agar gave substantial, but different, amounts of a family of four major trehalose-based LOSs. The core pentasaccharide LOS-I was a rhamnosyl diglucosyl-acylated trehalose. The heptasaccharide, LOS-II, was derived from LOS-I by adding xylose accompanied by a novel sugar (X); repeated addition of this sugar unit X gave the octasaccharide LOS-III. LOS-IV has a decasaccharide component with two additional unusual sugar units, YZ. In a recent study (Alexander, D. C., Jones, J. R., Tan, T., Chen, J. M., and Liu, J. (2004) J. Biol. Chem. 279, 18824-18833), chromatographically similar glycolipids were assigned to the family of phosphatidylinositol mannosides (PIMs) and a "PimF" (Rv1500) glycosyltransferase implicated in the conversion of a supposed "PIM5" to a "PIM7." The present study indicates that these putative PIMs are in fact members of the phosphorus-free LOS family of glycolipids and that the protein product of Rv1500, which we have now termed LosA, is a glycosyltransferase involved in transferring sugars to LOS-III to form LOS-IV of M. marinum. PMID- 16257959 TI - GSK-3beta directly phosphorylates and activates MARK2/PAR-1. AB - In Alzheimer disease (AD), the microtubule-associated protein tau is found hyperphosphorylated in paired helical filaments. Among many phosphorylated sites in tau, Ser-262 is the major site for abnormal phosphorylation of tau in AD brain. The kinase known to phosphorylate this particular site is MARK2, whose activation mechanism is yet to be studied. Our first finding that treatment of cells with LiCl, a selective inhibitor of another major tau kinase, glycogen synthase kinase-3beta (GSK-3beta), inhibits phosphorylation of Ser-262 of tau led us to investigate the possible involvement of GSK-3beta in MARK2 activation. In vitro kinase reaction revealed that recombinant GSK-3beta indeed phosphorylates MARK2, whereas it failed to phosphorylate Ser-262 of tau. Our further findings led us to conclude that GSK-3beta phosphorylates MARK2 on Ser-212, one of the two reported phosphorylation sites (Thr-208 and Ser-212) found in the activation loop of MARK2. Down-regulation of either GSK-3beta or MARK2 by small interfering RNAs suppressed the level of phosphorylation on Ser-262. These results, respectively, indicated that GSK-3beta is responsible for phosphorylating Ser-262 of tau through phosphorylation and activation of MARK2 and that the phosphorylation of tau at this particular site is predominantly mediated by a GSK-3beta-MARK2 pathway. These findings are of interest in the context of the pathogenesis of AD. PMID- 16257961 TI - Retention of mutant low density lipoprotein receptor in endoplasmic reticulum (ER) leads to ER stress. AB - Familial hypercholesterolemia is an autosomal dominant disease caused by mutations in the gene encoding the low density lipoprotein receptor (LDLR). More than 50% of these mutations lead to receptor proteins that are completely or partly retained in the endoplasmic reticulum (ER). The mechanisms involved in the intracellular processing and retention of mutant LDLR are poorly understood. In the present study we show that the G544V mutant LDLR associates with the chaperones Grp78, Grp94, ERp72, and calnexin in the ER of transfected Chinese hamster ovary cells. Retention of the mutant LDLR was shown to cause ER stress and activation of the unfolded protein response. We observed a marked increase in the activity of two ER stress sensors, IRE1 and PERK. These results show that retention of mutant LDLR in ER induces cellular responses, which might be important for the clinical outcome of familial hypercholesterolemia. PMID- 16257962 TI - Oxygen tension regulates mitochondrial DNA-encoded complex I gene expression. AB - Oxygen is a major regulator of nuclear gene expression. However, although mitochondria consume almost all of the O2 available to the cells, little is known about how O2 tension influences the expression of the mitochondrial genome. We show in O2-sensitive excitable rat PC12 cells that, among the mtDNA-encoded genes, hypoxia produced a specific down-regulation of the transcripts encoding mitochondrial complex I NADH dehydrogenase (ND) subunits, particularly ND4 and ND5 mRNAs and a stable mRNA precursor containing the ND5 and cytochrome b genes. This unprecedented effect of hypoxia was fast (developed in <30 min) and fairly reversible and occurred at moderate levels of hypoxia (O2 tensions in the range of 20-70 mm Hg). Hypoxic down-regulation of the mitochondrial complex I genes was paralleled by the reduction of complex I activity and was retarded by iron chelation, suggesting that an iron-dependent post-transcriptional mechanism could regulate mitochondrial mRNA stability. It is known that cell respiration is under tight control by the amount of proteins in mitochondrial complexes of the electron transport chain. Therefore, regulation of the expression of the mitochondrial (mtDNA)-encoded complex I subunits could be part of an adaptive mechanism to adjust respiration rate to the availability of O2 and to induce fast adaptive changes in hypoxic cells. PMID- 16257963 TI - Phosphorylation of WASP by the Cdc42-associated kinase ACK1: dual hydroxyamino acid specificity in a tyrosine kinase. AB - ACK1 is a nonreceptor tyrosine kinase that associates specifically with Cdc42. Relatively few ACK1 substrates and interacting proteins have been identified. In this study, we demonstrated that ACK1 phosphorylates the Wiskott-Aldrich syndrome protein (WASP), a Cdc42 effector that plays an important role in the formation of new actin filaments. ACK1 and WASP interact in intact cells, and overexpression of ACK1 promotes WASP phosphorylation. Phosphorylation of WASP in vitro was enhanced by the addition of Cdc42 or phosphatidylinositol 4,5-biphosphate, presumably due to release of the autoinhibitory interactions in WASP. Surprisingly, when we mapped the sites of WASP phosphorylation, we found that ACK1 possesses significant serine kinase activity toward WASP (directed at Ser 242), as well as tyrosine kinase activity directed at Tyr-256. A serine peptide derived from the Ser-242 WASP phosphorylation site is also a substrate for ACK1. ACK1 expressed in bacteria retained its serine kinase activity, eliminating the possibility of contamination with a copurifying kinase. Serine phosphorylation of WASP enhanced the ability of WASP to stimulate actin polymerization in mammalian cell lysates. Thus, the tyrosine kinase ACK1 acts as a dual specificity kinase toward this substrate. In contrast to other dual specificity kinases that more closely resemble Ser/Thr kinases, ACK1 is a tyrosine kinase with an active site that can accommodate both types of hydroxyamino acids in substrates. PMID- 16257964 TI - Differences in eNOS activity because of subcellular localization are dictated by phosphorylation state rather than the local calcium environment. AB - Nitric oxide (NO) produced in the endothelium via the enzyme endothelial nitric oxide synthase (eNOS) is an important vasoactive compound. Wild-type (WT) eNOS is localized to the plasma membrane and perinuclear/Golgi region by virtue of N terminal myristoylation and palmitoylation. Acylation-deficient mutants (G2AeNOS) remain cytosolic and release less NO in response to Ca2+-elevating agonists; a disparity that we hypothesized was attributed to the greater distance between G2AeNOS and plasma membrane Ca2+ influx channels. The reduced activity of G2AeNOS versus WT was reversed upon disruption of cellular integrity with detergents or sonication. NO production from both constructs relied almost exclusively on the influx of extracellular Ca2+, and elevating intracellular Ca2+ to saturating levels with 10 microM ionomycin in the presence of 10 mM extracellular Ca2+ equalized NO production. To identify the contribution of calcium to the differences in activity between these enzymes, we created Ca2+/CaM-independent eNOS mutants by deleting the two putative autoinhibitory domains of eNOS. There was no difference in NO production between WT and G2A-targeted Ca2+-independent eNOS, suggesting that Ca2+ was the factor responsible. When eNOS constructs were fused in-frame to the bioluminescent probe aequorin, membrane-bound probes were exposed to higher [Ca2+] in unstimulated cells but upon ionomycin stimulation, the probes experienced equal amounts of Ca2+. The WT and G2A enzymes displayed significant differences in the phosphorylation state of Ser617, Ser635, and Ser1179, and mutating all three sites to alanine or restoring phosphorylation with the phosphatase inhibitor calyculin abolished the differences in activity. We therefore conclude that the disparity in NO production between WTeNOS and G2AeNOS is not caused by different localized [Ca2+] upon stimulation with ionomycin, but rather differences in phosphorylation state between the two constructs. PMID- 16257965 TI - Enacyloxin IIa pinpoints a binding pocket of elongation factor Tu for development of novel antibiotics. AB - Elongation factor (EF-) Tu.GTP is the carrier of aminoacyl-tRNA to the programmed ribosome. Enacyloxin IIa inhibits bacterial protein synthesis by hindering the release of EF-Tu.GDP from the ribosome. The crystal structure of the Escherichia coli EF-Tu.guanylyl iminodiphosphate (GDPNP).enacyloxin IIa complex at 2.3 A resolution presented here reveals the location of the antibiotic at the interface of domains 1 and 3. The binding site overlaps that of kirromycin, an antibiotic with a structure that is unrelated to enacyloxin IIa but that also inhibits EF Tu.GDP release. As one of the major differences, the enacyloxin IIa tail borders a hydrophobic pocket that is occupied by the longer tail of kirromycin, explaining the higher binding affinity of the latter. EF-Tu.GDPNP.enacyloxin IIa shows a disordered effector region that in the Phe-tRNAPhe.EF-Tu (Thermus aquaticus).GDPNP.enacyloxin IIa complex, solved at 3.1 A resolution, is stabilized by the interaction with tRNA. This work clarifies the structural background of the action of enacyloxin IIa and compares its properties with those of kirromycin, opening new perspectives for structure-guided design of novel antibiotics. PMID- 16257967 TI - Integration requires a specific interaction of the donor DNA terminal 5'-cytosine with glutamine 148 of the HIV-1 integrase flexible loop. AB - Integration is essential for retroviral replication and gene therapy using retroviral vectors. Human immunodeficiency virus, type 1 (HIV-1), integrase specifically recognizes the terminal sequences of each long terminal repeat (LTR) and cleaves the 3'-end terminal dinucleotide 5'-GT. The exposed 3'-hydroxyl is then positioned for nucleophilic attack and subsequent strand transfer into another DNA duplex (target or chromosomal DNA). We report that both the terminal cytosine at the protruding 5'-end of the long terminal repeats (5'-C) and the integrase residue Gln-148 are critical for strand transfer. Proximity of the 5'-C and Gln-148 was demonstrated by disulfide cross-linking. Cross-linking is inhibited by the inhibitor 5CITEP 1-(5-chloroindol-3-yl)-3-hydroxy-3-(2H-tetrazol 5-yl)-propenone. We propose that strand transfer requires a conformational change of the integrase-viral (donor) DNA complex with formation of an H-bond between the N-3 of the 5'-C and the amine group of Gln-148. These findings have implications for the molecular mechanisms coupling 3'-processing and strand transfer as well as for the molecular pharmacology of integrase inhibitors. PMID- 16257968 TI - Identification of the fibroblast growth factor (FGF)-interacting domain in a secreted FGF-binding protein by phage display. AB - Fibroblast growth factor-binding proteins (FGF-BP) are secreted carrier proteins that release fibroblast growth factors (FGFs) from the extracellular matrix storage and thus enhance FGF activity. Here we have mapped the interaction domain between human FGF-BP1 and FGF-2. For this, we generated T7 phage display libraries of N-terminally and C-terminally truncated FGF-BP1 fragments that were then panned against immobilized FGF-2. From this panning, a C-terminal fragment of FGF-BP1 (amino acids 193-234) was identified as the minimum binding domain for FGF. As a recombinant protein, this C-terminal fragment binds to FGF-2 and enhances FGF-2-induced signaling in NIH-3T3 fibroblasts and GM7373 endothelial cells, as well as mitogenesis and chemotaxis of NIH-3T3 cells. The FGF interaction domain in FGF-BP1 is distinct from the heparin-binding domain (amino acids 110-143), and homology modeling supports the notion of a distinct domain in the C terminus that is conserved across different species. This domain also contains conserved positioning of cysteine residues with the Cys-214/Cys-222 positions in the human protein predicted to participate in disulfide bridge formation. Phage display of a C214A mutation of FGF-BP1 reduced binding to FGF-2, indicating the functional significance of this disulfide bond. We concluded that the FGF interaction domain is contained in the C terminus of FGF-BP1. PMID- 16257966 TI - M-RIP targets myosin phosphatase to stress fibers to regulate myosin light chain phosphorylation in vascular smooth muscle cells. AB - Vascular smooth muscle cell contraction and relaxation are directly related to the phosphorylation state of the regulatory myosin light chain. Myosin light chains are dephosphorylated by myosin phosphatase, leading to vascular smooth muscle relaxation. Myosin phosphatase is localized not only at actin-myosin stress fibers where it dephosphorylates myosin light chains, but also in the cytoplasm and at the cell membrane. The mechanisms by which myosin phosphatase is targeted to these loci are incompletely understood. We recently identified myosin phosphatase-Rho interacting protein as a member of the myosin phosphatase complex that directly binds both the myosin binding subunit of myosin phosphatase and RhoA and is localized to actin-myosin stress fibers. We hypothesized that myosin phosphatase-Rho interacting protein targets myosin phosphatase to the contractile apparatus to dephosphorylate myosin light chains. We used RNA interference to silence the expression of myosin phosphatase-Rho interacting protein in human vascular smooth muscle cells. Myosin phosphatase-Rho interacting protein silencing reduced the localization of the myosin binding subunit to stress fibers. This reduction in stress fiber myosin phosphatase-Rho interacting protein and myosin binding subunit increased basal and lysophosphatidic acid-stimulated myosin light chain phosphorylation. Neither cellular myosin phosphatase, myosin light chain kinase, nor RhoA activities were changed by myosin phosphatase-Rho interacting protein silencing. Furthermore, myosin phosphatase-Rho interacting protein silencing resulted in marked phenotypic changes in vascular smooth muscle cells, including increased numbers of stress fibers, increased cell area, and reduced stress fiber inhibition in response to a Rho-kinase inhibitor. These data support the importance of myosin phosphatase-Rho interacting protein-dependent targeting of myosin phosphatase to stress fibers for regulating myosin light chain phosphorylation state and morphology in human vascular smooth muscle cells. PMID- 16257969 TI - Mechanistic and functional studies of the interaction of a proline-rich antimicrobial peptide with mammalian cells. AB - Mammalian antimicrobial peptides provide rapid defense against infection by inactivating pathogens and by influencing the functions of cells involved in defense responses. Although the direct antibacterial properties of these peptides have been widely characterized, their multiple effects on host cells are only beginning to surface. Here we investigated the mechanistic and functional aspects of the interaction of the proline-rich antimicrobial peptide Bac7(1-35) with mammalian cells, as compared with a truncated analog, Bac7(5-35), lacking four critical N-terminal residues (RRIR) of the Bac7(1-35) sequence. By using confocal microscopy and flow cytometry, we showed that although the truncated analog Bac7(5-35) remains on the cell surface, Bac7(1-35) is rapidly taken up into 3T3 and U937 cells through a nontoxic energy- and temperature-dependent process. Cell biology-based assays using selective endocytosis inhibitors and spectroscopic and surface plasmon resonance studies of the interaction of Bac7(1-35) with phosphatidylcholine/cholesterol model membranes collectively suggest the concurrent contribution of macropinocytosis and direct membrane translocation. Structural studies with model membranes indicated that membrane-bound Bac7(5-35) is significantly more aggregated than Bac7(1-35) due to the absence of the N terminal cationic cluster, thus providing an explanation for hampered cellular internalization of the truncated form. Further investigations aimed to reveal functional implications of intracellular uptake of Bac7(1-35) demonstrated that it correlates with enhanced S phase entry of 3T3 cells, indicating a novel function for this proline-rich peptide. PMID- 16257970 TI - Mutations in the peroxin Pex26p responsible for peroxisome biogenesis disorders of complementation group 8 impair its stability, peroxisomal localization, and interaction with the Pex1p x Pex6p complex. AB - Peroxisome biogenesis disorders (PBDs) are fatal autosomal recessive diseases and are caused by impaired peroxisome biogenesis. PBDs are genetically heterogeneous and classified into 13 complementation groups (CGs). CG8 is one of the most common groups and has three clinical phenotypes, including Zellweger syndrome (ZS), neonatal adrenoleukodystrophy, and infantile Refsum disease (IRD). We recently isolated PEX26 as the pathogenic gene for PBD of CG8. Pex26p functions in recruiting to peroxisomes the complexes of the AAA ATPase peroxins, Pex1p and Pex6p. In the present work, we identified four distinct mutations in PEX26 from five patients of CG8 PBD including 2 with ZS and 3 with IRD, in addition to 7 mutant alleles in 8 patients in the first report describing the pathogenic PEX26 gene for CG8 PBD. Phenotype-genotype analyses revealed that temperature-sensitive (ts) peroxisome assembly gave rise to a milder IRD in contrast to the non-ts phenotype of the cells from ZS patients. Furthermore, we present several lines of evidence that show that the instability, insufficient binding to Pex1p x Pex6p complexes, or mislocalization of patient-derived Pex26p mutants is most likely responsible for the CG8 PBDs. PMID- 16257971 TI - Biochemical and mutational analyses of a unique clamp loader complex in the archaeon Methanosarcina acetivorans. AB - Clamp loaders orchestrate the switch from distributive to processive DNA synthesis. Their importance in cellular processes is underscored by their conservation across all forms of life. Here, we describe a new form of clamp loader from the archaeon Methanosarcina acetivorans. Unlike previously described archaeal clamp loaders, which are composed of one small subunit and one large subunit, the M. acetivorans clamp loader comprises two similar small subunits (M. acetivorans replication factor C small subunit (MacRFCS)) and one large subunit (MacRFCL). The relatedness of the archaeal and eukaryotic clamp loaders (which are made up of four similar small subunits and one large subunit) suggests that the M. acetivorans clamp loader may be an intermediate form in the archaeal/eukaryotic sister lineages. The clamp loader complex reconstituted from the three subunits MacRFCS1, MacRFCS2, and MacRFCL stimulated DNA synthesis by a cognate DNA polymerase in the presence of its sliding clamp. We used site directed mutagenesis in the Walker A and SRC motifs to examine the contribution of each subunit to the function of the M. acetivorans clamp loader. Although mutations in MacRFCL and MacRFCS2 did not impair clamp loading activity, any mutant clamp loader harboring a mutation in MacRFCS1 was devoid of the clamp loading property. Mac-RFCS1 is therefore critical to the clamp loading activity of the M. acetivorans clamp loader. It is our anticipation that the discovery of this unique replication factor C homolog will lead to critical insights into the evolution of more complex clamp loaders from simpler ones as more complex organisms evolved in the archaeal/eukaryotic sister lineages. PMID- 16257972 TI - TRP-ML1 is a lysosomal monovalent cation channel that undergoes proteolytic cleavage. AB - Mutations in the gene MCOLN1 coding for the TRP (transient receptor potential) family ion channel TRP-ML1 lead to the lipid storage disorder mucolipidosis type IV (MLIV). The function and role of TRP-ML1 are not well understood. We report here that TRP-ML1 is a lysosomal monovalent cation channel. Both native and recombinant TRP-ML1 are cleaved resulting in two products. Recombinant TRP-ML1 is detected as the full-length form and as short N- and C-terminal forms, whereas in native cells mainly the cleaved N and C termini are detected. The N- and C terminal fragments of TRP-ML1 were co-immunoprecipitated from cell lysates and co eluted from a Ni2+ column. TRP-ML1 undergoes proteolytic cleavage that is inhibited by inhibitors of cathepsin B (CatB) and is altered when TRP-ML1 is expressed in CatB-/- cells. N-terminal sequencing of purified C-terminal fragment of TRP-ML1 expressed in Sf9 cells indicates a cleavage site at Arg200 downward arrow Pro201. Consequently, the conserved R200H mutation changed the cleavage pattern of TRP-ML1. The cleavage inhibited TRP-ML1 channel activity. This work provides the first example of inactivation by cleavage of a TRP channel. The significance of the cleavage to the function of TRP-ML1 is under investigation. PMID- 16257973 TI - The RNA binding G-patch domain in retroviral protease is important for infectivity and D-type morphogenesis of Mason-Pfizer monkey virus. AB - Retroviral proteases (PRs) cleave the viral polyprotein precursors into functional mature proteins late during particle release and are essential for viral replication. Unlike most retroviruses, beta-retroviruses, including Mason Pfizer monkey virus (M-PMV), assemble immature capsids within the cytoplasm of the cell. The activation of beta-retroviral proteases must be highly regulated, because processing of the Gag-related polyprotein precursors occurs only after transport of immature capsids to the plasma membrane and budding. Several beta retroviral proteases have unique C-terminal extension sequences, containing a glycine-rich motif (G-patch), which specifically binds in vitro to single stranded nucleic acids. In M-PMV PR the G-patch is removed in vitro as well as in vivo by autoproteolytic processing to yield truncated active forms of PR. To investigate the role of the G-patch domain on the virus life cycle, we introduced mutations within the C-terminal domain of protease. We found that the G-patch domain of M-PMV PR is not required for the processing of viral polyproteins, but it significantly influences the infectivity of M-PMV, the activity of reverse transcriptase, and assembly of immature capsid within the cells. These results demonstrate for the first time that the G-patch domain of M-PMV PR is critical for the life cycle of beta-retroviruses, and its evolutionary conservation within members of this genus suggests its importance for retroviruses that display D type morphology. PMID- 16257974 TI - The survival kinase Mirk/dyrk1B is activated through Rac1-MKK3 signaling. AB - The serine/threonine kinase Mirk/dyrk1B is activated in several solid tumors where it mediates cell survival, but the mechanism by which Mirk is activated in tumors is unknown. We now demonstrate that Mirk is activated as a kinase by signaling from Rac1 to the mitogen-activated protein kinase kinase MKK3. Rac is a Ras superfamily GTPase that, when activated, functions downstream of Ras oncoproteins to promote cell survival, transformation, and membrane ruffling. The constitutively active mutant Rac1QL activated Mirk in several cell types through MKK3, which in turn activated Mirk by phosphorylation. Dominant negative Rac1, dominant negative MKK3, and knockdown of MKK3 by RNA interference inhibited the kinase activity of co-expressed Mirk. E-cadherin ligation in confluent Madin Darby canine kidney (MDCK) epithelial cells is known to transiently activate Rac1. Mirk was activated by endogenous Rac1 following E-cadherin ligation in confluent MDCK epithelial cells, whereas treatment of confluent MDCK cells with an Rac1 inhibitor decreased Mirk activity. Disruption of cadherin ligation by EGTA or prevention of cadherin ligation by maintenance of cells at subconfluent density blocked activation of Mirk. Engagement of cadherin molecules on subconfluent cells by an E-cadherin/Fc chimeric molecule transiently activated both Rac1 and Mirk with a similar time course. Rac activity is up-regulated in many human tumors and mediates survival signals, which enable tumor cells to evade apoptosis. This study characterizes a new anti-apoptotic signaling pathway that connects Rac1 with a novel downstream effector, Mirk kinase, which has recently been demonstrated to mediate survival in human tumors. PMID- 16257975 TI - The conserved Leu-724 residue is required for both serine phosphorylation and co activator recruitment for Stat1-mediated transcription activation in response to interferon-gamma. AB - The signal transducer and activator of transcription (STAT) proteins, a family of latent cytoplasmic transcription factors, become activated in response to extracellular ligand binding to cell surface receptors through tyrosine phosphorylation. Concurrently, a serine phosphorylation event in the transcription activation domain (serine 727 for Stat1) occurs. This serine phosphorylation is essential for the maximal transcription activity of Stat1. Here we show that, in addition to the Ser-727 residue and its phosphorylation, the conserved Leu-724 residue is also essential for gene activation mediated by Stat1. When Leu-724 is mutated to Ala, phosphorylation of Stat1 Ser-727 is defective both in vivo and in vitro. Surprisingly, we found a StatL724I mutant that lacks transcription activity despite normal Ser-727 phosphorylation. Further analyses show that Leu-724, as well as the phospho-Ser-727, are essential for the recruitment of the transcription co-activator CBP/p300 to the promoters of Stat1 target genes. Our results demonstrate that the conserved Leu-724 residue is a key residue that controls the maximal transcription activities of Stat1 in IFN-gamma signaling. PMID- 16257976 TI - Analysis of RovA, a transcriptional regulator of Yersinia pseudotuberculosis virulence that acts through antirepression and direct transcriptional activation. AB - The transcription factor RovA of Yersinia pseudotuberculosis and analogous proteins in other Enterobacteriaceae activate the expression of virulence genes that play a crucial role in stress adaptation and pathogenesis. In this study, we demonstrate that the RovA protein forms dimers independent of DNA binding, stimulates RNA polymerase, most likely via its C-terminal domain, and counteracts transcriptional repression by the histone-like protein H-NS. As the molecular function of the RovA family is largely uncharacterized, random mutagenesis and terminal deletions were used to identify functionally important domains. Our analysis showed that a winged-helix motif in the center of the molecule is essential and directly involved in DNA binding. Terminal deletions and amino acid changes within both termini also abrogate RovA activation and DNA-binding functions, most likely due to their implication in dimer formation. Finally, we show that the last four amino acids of RovA are crucial for activation of gene transcription. Successive deletions of these residues result in a continuous loss of RovA activity. Their removal reduced the capacity of RovA to activate RNA polymerase and abolished transcription of RovA-activated promoters in the presence of H-NS, although dimerization and DNA binding functions were retained. Our structural model implies that the final amino acids of RovA play a role in protein-protein interactions, adjusting RovA activity. PMID- 16257977 TI - Position of the fluorescent label is a crucial factor determining signal intensity in microarray hybridizations. AB - A key issue in applications of short oligonucleotide-based microarrays is how to design specific probes with high sensitivity. Some details of the factors affecting microarray hybridization remain unclear, hampering a reliable quantification of target nucleic acids. We have evaluated the effect of the position of the fluorescent label [position of label (POL)] relative to the probe target duplex on the signal output of oligonucleotide microarrays. End-labelled single-stranded DNA targets of different lengths were used for hybridization with perfect-match oligonucleotide probe sets targeting different positions of the same molecule. Hybridization results illustrated that probes targeting the labelled terminus of the target showed significantly higher signals than probes targeting other regions. This effect was independent of the target gene, the fluorophore and the slide surface chemistry. Comparison of microarray signal patterns of fluorescently end-labelled, fluorescently internally random-labelled and radioactively end-labelled target-DNAs with the same set of oligonucleotide probes identified POL as a critical factor affecting signal intensity rather than binding efficiency. Our observations define a novel determinant for large differences of signal intensities. Application of the POL effect may contribute to better probe design and data interpretation in microarray applications. PMID- 16257978 TI - A method to find tissue-specific novel sites of selective adenosine deamination. AB - Site-selective adenosine (A) to inosine (I) RNA editing by the ADAR enzymes has been found in a variety of metazoan from fly to human. Here we describe a method to detect novel site-selective A to I editing that can be used on various tissues as well as species. We have shown previously that there is a preference for ADAR2 binding to selectively edited sites over non-specific interactions with random sequences of double-stranded RNA. The method utilizes immunoprecipitation (IP) of intrinsic RNA-protein complexes to extract substrates subjected to site-selective editing in vivo, in combination with microarray analyses of the captured RNAs. We show that known single sites of A to I editing can be detected after IP using an antibody against the ADAR2 protein. The RNA substrates were verified by RT-PCR, RNase protection and microarray. Using this method it is possible to uniquely identify novel single sites of selective A to I editing. PMID- 16257979 TI - A colorimetric method for point mutation detection using high-fidelity DNA ligase. AB - The present study reported proof-of-principle for a genotyping assay approach that can detect single nucleotide polymorphisms (SNPs) through the gold nanoparticle assembly and the ligase reaction. By incorporating the high-fidelity DNA ligase (Tth DNA ligase) into the allele-specific ligation-based gold nanoparticle assembly, this assay provided a convenient yet powerful colorimetric detection that enabled a straightforward single-base discrimination without the need of precise temperature control. Additionally, the ligase reaction can be performed at a relatively high temperature, which offers the benefit for mitigating the non-specific assembly of gold nanoparticles induced by interfering DNA strands. The assay could be implemented via three steps: a hybridization reaction that allowed two gold nanoparticle-tagged probes to hybrid with the target DNA strand, a ligase reaction that generates the ligation between perfectly matched probes while no ligation occurred between mismatched ones and a thermal treatment at a relatively high temperature that discriminate the ligation of probes. When the reaction mixture was heated to denature the formed duplex, the purple color of the perfect-match solution would not revert to red, while the mismatch gave a red color as the assembled gold nanoparticles disparted. The present approach has been demonstrated with the identification of a single-base mutation in codon 12 of a K-ras oncogene that is of significant value for colorectal cancers diagnosis, and the wild-type and mutant type were successfully scored. To our knowledge, this was the first report concerning SNP detection based on the ligase reaction and the gold nanoparticle assembly. Owing to its ease of operation and high specificity, it was expected that the proposed procedure might hold great promise in practical clinical diagnosis of gene-mutant diseases. PMID- 16257980 TI - Correlation between sequence conservation and the genomic context after gene duplication. AB - A key complication in comparative genomics for reliable gene function prediction is the existence of duplicated genes. To study the effect of gene duplication on function prediction, we analyze orthologs between pairs of genomes where in one genome the orthologous gene has duplicated after the speciation of the two genomes (i.e. inparalogs). For these duplicated genes we investigate whether the gene that is most similar on the sequence level is also the gene that has retained the ancestral gene-neighborhood. Although the majority of investigated cases show a consistent pattern between sequence similarity and gene-neighborhood conservation, a substantial fraction, 29-38%, is inconsistent. The observation of inconsistency is not the result of a chance outcome owing to a lack of divergence time between inparalogs, but rather it seems to be the result of a chance outcome caused by very similar rates of sequence evolution of both inparalogs relative to their ortholog. If one-to-one orthologous relationships are required, it is advisable to combine contextual information (i.e. gene-neighborhood in prokaryotes and co-expression in eukaryotes) with protein sequence information to predict the most probable functional equivalent ortholog in the presence of inparalogs. PMID- 16257981 TI - 8-methyl-2'-deoxyguanosine incorporation into parallel DNA quadruplex structures. AB - This paper concerns the Circular Dichroism (CD) and Nuclear Magnetic Resonance (NMR) structural studies of the quadruple helix arrangements adopted by three tailored oligodeoxyribonucleotide analogues, namely d(TG(Me)GGT), d(TGG(Me)GT) and d(TGGG(Me)T), where dG(Me) represents a 8-methyl-2'-deoxyguanosine residue. The results of this study clearly demonstrate that the effects of the incorporation of dG(Me) instead of a dG residue are strongly dependant upon the positioning of a single base replacement along the sequence. As such, d(TG(Me)GGT), d(TGG(Me)GT) have been found to form 4-fold symmetric quadruplexes with all strands parallel and equivalent to each other, each more stable than their natural counterpart. NMR experiments clearly indicate that [d(TG(Me)GGT)]4 possesses a G(Me)-tetrad with all dG(Me) residues in a syn-glycosidic conformation while an anti-arrangement is apparent for the four dG(Me) of [d(TGG(Me)GT)]4. As the two complexes show a quite different CD behaviour, a possible relationship between the presence of residues adopting syn-glycosidic conformations and CD profiles is briefly discussed. As far as d(TGGG(Me)T) is concerned, NMR data indicate that at 25 degrees C it exists primarily as a single strand conformation in equilibrium with minor amounts of a quadruplex structure. PMID- 16257984 TI - Robust multi-scale clustering of large DNA microarray datasets with the consensus algorithm. AB - MOTIVATION: Hierarchical and relocation clustering (e.g. K-means and self organizing maps) have been successful tools in the display and analysis of whole genome DNA microarray expression data. However, the results of hierarchical clustering are sensitive to outliers, and most relocation methods give results which are dependent on the initialization of the algorithm. Therefore, it is difficult to assess the significance of the results. We have developed a consensus clustering algorithm, where the final result is averaged over multiple clustering runs, giving a robust and reproducible clustering, capable of capturing small signal variations. The algorithm preserves valuable properties of hierarchical clustering, which is useful for visualization and interpretation of the results. RESULTS: We show for the first time that one can take advantage of multiple clustering runs in DNA microarray analysis by collecting re-occurring clustering patterns in a co-occurrence matrix. The results show that consensus clustering obtained from clustering multiple times with Variational Bayes Mixtures of Gaussians or K-means significantly reduces the classification error rate for a simulated dataset. The method is flexible and it is possible to find consensus clusters from different clustering algorithms. Thus, the algorithm can be used as a framework to test in a quantitative manner the homogeneity of different clustering algorithms. We compare the method with a number of state-of the-art clustering methods. It is shown that the method is robust and gives low classification error rates for a realistic, simulated dataset. The algorithm is also demonstrated for real datasets. It is shown that more biological meaningful transcriptional patterns can be found without conservative statistical or fold change exclusion of data. AVAILABILITY: Matlab source code for the clustering algorithm ClusterLustre, and the simulated dataset for testing are available upon request from T.G. and O.W. PMID- 16257983 TI - SegH and Hef: two novel homing endonucleases whose genes replace the mobC and mobE genes in several T4-related phages. AB - T4 contains two groups of genes with similarity to homing endonucleases, the seg genes (similarity to endonucleases encoded by group I introns) containing GIY-YIG motifs and the mob-genes (similarity to mobile endonucleases) containing H-N-H motifs. The four seg-genes characterized to date encode homing endonucleases with cleavage sites close to their respective gene loci while none of the mob-genes have been shown to cleave DNA. Of 18 phages screened, only T4 was found to have mobC while mobE genes were found in five additional phages. Interestingly, three phages encoded a seg-like gene (hereby called segH) with a GIY-YIG motif in place of mobC. An additional phage has an unrelated gene called hef (homing endonuclease-like function) in place of the mobE gene. The gene products of both novel genes displayed homing endonuclease activity with cleavage site specificity close to their respective genes. In contrast to intron encoded homing endonucleases, both SegH and Hef can cleave their own DNA as well as DNA from phages without the genes. Both segH and mobE (and most likely hef) can home between phages in mixed infections. We discuss why it might be a selective advantage for phage freestanding homing endonucleases to cleave both HEG containing and HEG-less genomes. PMID- 16257982 TI - Mutagenic effects of abasic and oxidized abasic lesions in Saccharomyces cerevisiae. AB - 2-deoxyribonolactone (L) and 2-deoxyribose (AP) are abasic sites that are produced by ionizing radiation, reactive oxygen species and a variety of DNA damaging agents. The biological processing of the AP site has been examined in the yeast Saccharomyces cerevisiae. However, nothing is known about how L is processed in this organism. We determined the bypass and mutagenic specificity of DNA containing an abasic site (AP and L) or the AP analog tetrahydrofuran (F) using an oligonucleotide transformation assay. The tetrahydrofuran analog and L were bypassed at 10-fold higher frequencies than the AP lesions. Bypass frequencies of lesions were greatly reduced in the absence of Rev1 or Polzeta (rev3 mutant), but were only marginally reduced in the absence of Poleta (rad30 mutant). Deoxycytidine was the preferred nucleotide inserted opposite an AP site whereas dA and dC were inserted at equal frequencies opposite F and L sites. In the rev1 and rev3 strains, dA was the predominant nucleotide inserted opposite these lesions. Overall, we conclude that both Rev1 and Polzeta are required for the efficient bypass of abasic sites in yeast. PMID- 16257985 TI - A generic motif discovery algorithm for sequential data. AB - MOTIVATION: Motif discovery in sequential data is a problem of great interest and with many applications. However, previous methods have been unable to combine exhaustive search with complex motif representations and are each typically only applicable to a certain class of problems. RESULTS: Here we present a generic motif discovery algorithm (Gemoda) for sequential data. Gemoda can be applied to any dataset with a sequential character, including both categorical and real valued data. As we show, Gemoda deterministically discovers motifs that are maximal in composition and length. As well, the algorithm allows any choice of similarity metric for finding motifs. Finally, Gemoda's output motifs are representation-agnostic: they can be represented using regular expressions, position weight matrices or any number of other models for any type of sequential data. We demonstrate a number of applications of the algorithm, including the discovery of motifs in amino acids sequences, a new solution to the (l,d)-motif problem in DNA sequences and the discovery of conserved protein substructures. AVAILABILITY: Gemoda is freely available at http://web.mit.edu/bamel/gemoda PMID- 16257986 TI - Classification using functional data analysis for temporal gene expression data. AB - MOTIVATION: Temporal gene expression profiles provide an important characterization of gene function, as biological systems are predominantly developmental and dynamic. We propose a method of classifying collections of temporal gene expression curves in which individual expression profiles are modeled as independent realizations of a stochastic process. The method uses a recently developed functional logistic regression tool based on functional principal components, aimed at classifying gene expression curves into known gene groups. The number of eigenfunctions in the classifier can be chosen by leave-one out cross-validation with the aim of minimizing the classification error. RESULTS: We demonstrate that this methodology provides low-error-rate classification for both yeast cell-cycle gene expression profiles and Dictyostelium cell-type specific gene expression patterns. It also works well in simulations. We compare our functional principal components approach with a B spline implementation of functional discriminant analysis for the yeast cell cycle data and simulations. This indicates comparative advantages of our approach which uses fewer eigenfunctions/base functions. The proposed methodology is promising for the analysis of temporal gene expression data and beyond. AVAILABILITY: MATLAB programs are available upon request. PMID- 16257987 TI - Intelligent client for integrating bioinformatics services. AB - MOTIVATION: In addition to existing bioinformatics software, a lot of new tools are being developed world wide to supply services for an ever growing, widely dispersed and heterogeneous collection of biological data. The integration of these resources under a common platform is a challenging task. To this end, several groups are developing integration technologies, in which services are usually registered in some sort of catalogue to allow novel discovering and accessing mechanisms to be implemented. However, each service demands specific interfaces to accommodate their parameters and it is a complicated task linking the different service inputs and outputs to solve a biological problem. RESULTS: In this work we address the design and implementation of a versatile web client to access BioMOBY compatible services (a system by which a client can interact with multiple sources of biological data regardless of the underlying format or schema) using the service description stored in the BioMOBY catalogue. The automatic interface generator significantly reduces developing time and produces uniform service access mechanisms. The design and proof of concept (for such a client) including the generic interface generator have been developed and implemented in the National Institute for Bioinformatics in Spain. AVAILABILITY: The INB (National Institute for Bioinformatics, Spain) platform is available at www.inab.org/MOWServ PMID- 16257989 TI - Development of a questionnaire for assessment of the psychosocial impact of dental aesthetics in young adults. AB - The aim of this study was to develop a psychometric instrument for assessment of orthodontic-specific aspects of quality of life. The study subjects, 194 young adults aged 18-30 years, were interviewed using a pool of 23 items dealing with the psychosocial impact of dental aesthetics. Self- and interviewer-rating of the dental aesthetic appearance of each subject were carried out using the Aesthetic Component (AC) of the Index of Orthodontic Treatment Need (IOTN). Additionally, the Perception of Occlusion Scale and a modification of the Dental Aesthetic Index (DAI) were applied. Factorial analyses identified four measures within the item pool, namely Dental Self-Confidence, Social Impact, Psychological Impact, and Aesthetic Concern. The factor structure was confirmed in an independent sample of 83 subjects aged 18-33 years. The reliabilities of the factor analysis derived scales were between alpha (alpha) 0.85 and 0.91. They differed between respondents with varying severity of malocclusion, as assessed by subject and interviewer ratings. The results suggest that the proposed instrument, termed the 'Psychosocial Impact of Dental Aesthetics Questionnaire' (PIDAQ), meets the criteria of factorial stability across samples and criterion-related validity and reliability, and might be a promising tool for further research and clinical application in orthodontics. PMID- 16257988 TI - A comparative study of dental arch widths: extraction and non-extraction treatment. AB - The aim of this study was to determine the pre- and post-treatment width changes in the canine, premolar and molar regions in subjects treated with extraction of four first premolars, non-extraction, and non-extraction with rapid maxillary expansion (RME). Pre- and post-treatment orthodontic study models of 60 females (13.83 +/- 2.77 years) and 24 males (14.33 +/- 2.67 years) who underwent comprehensive orthodontic therapy were evaluated. Forty-two were treated non extraction with fixed appliance therapy, 15 non-extraction with RME, and 27 with extraction of the first premolars. In addition to standard descriptive statistical calculations, one way ANOVA was used for comparison of the groups, and the post hoc Tukey multiple comparison test for comparison of the subgroups. The results revealed that the distance between the upper canines was not affected by the treatment modality. Upper premolar and molar arch widths increased more in the non-extraction subjects when compared with those with extractions, with the greatest increase in patients with RME. In the lower canine area the extraction group showed the widest arch width at the end of treatment. There was also a 0.60 mm decrease in the lower canine width in the non-extraction group. A decrease was found in lower inter-premolar and molar distances due to consolidation of the extraction spaces. When making a decision between non-extraction with maxillary expansion and extraction treatment modalities in borderline cases where there is constriction in the upper inter-premolar distance, apart from taking profile values into consideration, it should be borne in mind that expansion treatment can be helpful in achieving a wider arch form. PMID- 16257990 TI - Temporomandibular joint internal derangement in mandibular asymmetry. What is the relationship? AB - The purpose of this study was to clarify the contributing factors and their influences on temporomandibular joint internal derangement (TMJ ID) symptoms in subjects with mandibular asymmetry. Postero-anterior (PA) cephalograms of 187 pre orthodontic treatment subjects (aged 18-45 years, mean 23.9 years) were used to investigate the inclination of the frontal occlusal (FOP) and frontal mandibular (FMP) planes to determine vertical asymmetry. Mandibular dental midline shift (DMS) and mandibular midline shift (MMS) were studied to determine transverse asymmetry. The degree of asymmetry was analysed in conjunction with the results from self-administered TMJ ID history forms. A prevalence of TMJ ID was most related to the inclination of the FMP (P < 0.01), with the symptoms being notably higher when the cant was greater than 3 degrees. The symptomatic side was related only to the inclination of the FOP and FMP. Symptoms confined to the ipsilateral side were primarily found in subjects with mild asymmetry, whereas symptoms on both sides and those on the contralateral side were greater in those with moderate and severe asymmetry, respectively. No significant correlation was found for DMS and MMS. The results suggest that the degree of asymmetry in the vertical dimension is significantly correlated with TMJ ID symptoms. PMID- 16257991 TI - Craniofacial morphology in Chinese female twins: a semi-longitudinal cephalometric study. AB - It would be of benefit to have a better understanding of the relative effects of genetics and environmental factors on craniofacial parameters when undertaking orthodontic therapy and treatment planning. However, there is a lack of such information in pre-adolescents. The aim of this study was to verify the degree of genetic and environmental contribution to the growth of the facial skeleton in twins aged 6 to 12 years. The material comprised the lateral cephalograms of 89 pairs of female twins in Beijing, China, of whom 61 pairs were diagnosed by DNA analysis as monozygotic (MZ) and 28 pairs as dizygotic (DZ). Four main groups (with a starting age of 6, 7, 9 and 11 years) were studied in a semi-longitudinal manner, with a sub-group further investigated for 2-4 consecutive years. The total sample therefore consisted of 183 pairs (MZ 110, DZ 73) aged from 6 to 12 years. The depths of the cranial base, mid and lower face were measured, as well as anterior and posterior face height. A two-tailed t-test showed significant environmental effects on lower face depth (P < 0.01), whilst genetic effects on face height were also significant (P < 0.01). The results suggest that early orthodontic intervention would have a greater influence on the antero-posterior rather than on the vertical plane of growth. PMID- 16257992 TI - Improvements in quality of clinical care in English general practice 1998-2003: longitudinal observational study. AB - OBJECTIVE: To measure changes in quality of care for three major chronic diseases (coronary heart disease, asthma, and type 2 diabetes) between 1998 and 2003. DESIGN: Longitudinal cohort study. SETTING: 42 general practices in six geographical areas of England (Avon, Bury/Rochdale, Enfield, Oldham, Somerset, South Essex). PARTICIPANTS: Medical record data for 2300 patients with diabetes, asthma, or coronary heart disease in 1998, and 1495 patients in 2003. MAIN OUTCOME MEASURE: Quality of care assessed against predefined evidence based review criteria. RESULTS: Between 1998 and 2003, quality of care improved markedly in terms of maximum possible scores on the review criteria, from 60.5% to 78.1% for coronary heart disease (change = 17.6, 95% confidence interval 13.9 to 21.4; P < 0.001), 60.1% to 70.3% for asthma (10.2, 4.6 to 15.8; P = 0.001), and 70.4% to 77.7% for diabetes (7.3, 3.5 to 11.1; P = 0.001). Important changes occurred to several indicators potentially related to improved health outcomes. These included improved control of serum cholesterol (to < or = 5 mmol/l) from 17.6% to 61.4% in coronary heart disease and from 21.5% to 52% in diabetes and control of blood pressure to < or = 150/90 in coronary heart disease from 47.3% to 72.2% and to < or = 145/85 in diabetes from 21.8% to 35.8%. A small, non significant improvement in glycaemic control occurred among diabetic patients (37.9% to 39.7% with HbA1c < 7.4%). Significant improvements also occurred in the recording of exercise capacity and diet and weight advice for patients with coronary heart disease; of smoking advice, peak flow, and symptoms for patients with asthma; and of creatinine, weight, and HbA(1c) for patients with diabetes. Over the five years, more improvement in coronary heart disease care occurred in large practices and practices in affluent areas. CONCLUSIONS: Substantial improvements were seen in quality of care for the three conditions studied between 1998 and 2003, a time of systematic quality improvement initiatives in the NHS. The changes were most marked for coronary heart disease. English general practices could be expected to achieve high clinical quality scores in the initial year of a new contact, which provides financial incentives for high quality care from 2004. PMID- 16257993 TI - Depression and unwanted first pregnancy: longitudinal cohort study. AB - OBJECTIVE: To examine the outcomes of an unwanted first pregnancy (abortion v live delivery) and risk of depression and to explain discrepancies with previous research that used the same dataset. DESIGN: Longitudinal cohort study. SETTING: Nationally representative sample of US men and women aged 14-24 in 1979. PARTICIPANTS: 1247 women in the US national longitudinal survey of youth who aborted or delivered an unwanted first pregnancy. MAIN OUTCOME MEASURES: Clinical cut-off and continuous scores on a 1992 measure of the Center for Epidemiological Studies depression scale. RESULTS: Terminating compared with delivering an unwanted first pregnancy was not directly related to risk of clinically significant depression (odds ratio 1.19, 95% confidence interval 0.85 to 1.66). No evidence was found of a relation between pregnancy outcome and depression in analyses of subgroups known to vary in under-reporting of abortion. In analyses of the characteristics of non-respondents, refusal to provide information on abortion did not explain the lack of detecting a relation between abortion and mental health. The abortion group had a significantly higher mean education and income and lower total family size, all of which were associated with a lower risk of depression. CONCLUSIONS: Evidence that choosing to terminate rather than deliver an unwanted first pregnancy puts women at higher risk of depression is inconclusive. Discrepancies between current findings and those of previous research using the same dataset primarily reflect differences in coding of a first pregnancy. PMID- 16257994 TI - Evaluation of costs and effects of epidural analgesia and patient-controlled intravenous analgesia after major abdominal surgery. AB - BACKGROUND: The outcome of different treatment strategies for postoperative pain has been an issue of controversy. Apart from efficacy and effectiveness a policy decision should also consider cost-effectiveness. Since economic analyses on postoperative pain treatment are rare we developed a decision model in a pilot cost-effectiveness analysis (CEA) comparing epidural analgesia (EDA) and patient controlled intravenous analgesia (PCIA) after major abdominal surgery in routine care. METHODS: Using a decision-tree model, treatment with EDA (ropivacaine and morphine) was compared with PCIA (morphine). Effects and costs of treatment were established. The number of pain-free days at rest (pain intensity <30 using visual analogue scale 1-100 mm) was the primary measure of effect. An incremental cost-effectiveness ratio (ICER) was calculated as the difference in direct costs divided by the difference in effect. A database on 644 patients collected for the purpose of quality control during the period of 1997 to 1999 was the main data source. Sensitivity analysis was used to test uncertain data. RESULTS: EDA was more effective in terms of pain-free days but more expensive. The additional cost for each pain-free day was 5652 Euros. CONCLUSION: It is a judgement of value if the additional cost is reasonable. When the cost of around 55,000 Euros per gained life-year with full health for other interventions is debated, our result indicates poor cost-effectiveness for EDA. Before any conclusion can be drawn concerning policy recommendations the difference in costs has to be related to other outcome measures as length of hospital stay, morbidity and mortality are required. PMID- 16257995 TI - Airway management of patients with tracheobronchial stents. AB - The use of tracheobronchial stents for compromised large airways is increasing. We provide a case series highlighting some of the complications of airway management in patients with tracheobronchial stents in situ and propose an approach for dealing with this potentially complicated situation. PMID- 16257996 TI - Assessment of pulse transit time to indicate cardiovascular changes during obstetric spinal anaesthesia. AB - BACKGROUND: Pulse transit time (PTT) measurement may provide rapidly available beat-to-beat cardiovascular information when conditions change quickly and routine invasive arterial pressure measurement is not justified, for example during obstetric spinal anaesthesia. Method. We obtained ethics approval for an observational study of PTT during the onset of spinal anaesthesia in patients having elective or urgent Caesarean section. PTT was measured as the difference in time between the peak of the ECG R wave and the upstroke of the toe plethysmograph. Arterial pressure was measured by non-invasive sphygmomanometry. RESULTS: We analysed data from 58 normotensive patients and 15 patients with pregnancy-induced hypertension (PIH). PTT increased with the onset of spinal anaesthesia as arterial pressure decreased. An increase of 20% in PTT was 74% sensitive and 70% specific in indicating a decrease in mean arterial pressure of more than 10%. Changes in PTT were related to changes in mean arterial pressure (r2=0.55, P<0.0001). Arterial pressure changes were greater and PTT increased significantly more quickly in the normotensive patients than in the patients with hypertension [median, quartiles: 32 (14, 56) ms min(-1) compared with 7 (6, 18) ms min(-1); P<0.01, Mann-Whitney U-test]. However, the relationship between PTT and arterial pressure was similar for the normotensive patients and the patients with PIH. CONCLUSION: PTT measurement gave a beat-to-beat indication of arterial pressure during spinal anaesthesia, and could be developed to allow prediction of the onset of hypotension. PMID- 16257998 TI - All-trans retinoic acid modulates radiation-induced proliferation of lung fibroblasts via IL-6/IL-6R system. AB - Although high-dose thoracic radiotherapy is an effective strategy for some malignancies including lung cancers and malignant lymphomas, it often causes complications of radiation fibrosis. To study the mechanism initiating tissue fibrosis, we investigated irradiation-induced cytokine production from human lung fibroblastic cells and found that IL-6 production was stimulated by irradiation. IL-6 is an autocrine growth factor for human myeloma cells, and retinoic acid is reported to inhibit their growth. Thus we evaluated the effect of all-trans retinoic acid (ATRA) on cell proliferation of lung fibroblasts along with the cytokine/receptor system. Irradiation-dependent stimulation of IL-6 production was correlated with increased NF-kappaB activity, and ATRA reduced this effect. Irradiation also increased the levels of mRNA for IL-6R and gp130, which were blocked by coexisting ATRA. Furthermore, IL-6 stimulated cell proliferation in dose-dependent manner but was overcome by pharmacological concentration of ATRA. These effects of ATRA were inhibited by rottlerin, which suggests ATRA abolished irradiation-induced stimulation through a PKCdelta-dependent pathway. Finally, we demonstrated that IL-6 transcripts in the lung were upregulated at 2 mo after irradiation, and the effect was inhibited by the intraperitoneal administration of ATRA. ATRA is expected to have an advantage for radiotherapy in its antitumor effects, as reported previously, and to prevent radiotherapy-induced pulmonary injury. PMID- 16257997 TI - Inhibition of phosphodiesterase 4 amplifies cytokine-dependent induction of arginase in macrophages. AB - Arginase is greatly elevated in asthma and is thought to play a role in the pathophysiology of this disease. As inhibitors of phosphodiesterase 4 (PDE4), the predominant PDE in macrophages, elevate cAMP levels and reduce inflammation, they have been proposed for use in treatment of asthma and chronic obstructive pulmonary disease. As cAMP is an inducer of arginase, we tested the hypothesis that a PDE4 inhibitor would enhance macrophage arginase induction by key cytokines implicated in asthma and other pulmonary diseases. RAW 264.7 cells were stimulated with IL-4 or transforming growth factor (TGF)-beta, with and without the PDE4 inhibitor rolipram. IL-4 and TGF-beta increased arginase activity 16- and 5-fold, respectively. Rolipram alone had no effect but when combined with IL 4 and TGF-beta synergistically enhanced arginase activity by an additional 15- and 5-fold, respectively. The increases in arginase I protein and mRNA levels mirrored increases in arginase activity. Induction of arginase II mRNA was also enhanced by rolipram but to a much lesser extent than arginase I. Unlike its effect in RAW 264.7 cells, IL-4 alone did not increase arginase activity in human alveolar macrophages (AM) from healthy volunteers. However, combining IL-4 with agents to induce cAMP levels induced arginase activity in human AM significantly above the level obtained with cAMP-inducing agents alone. In conclusion, agents that elevate cAMP significantly enhance induction of arginase by cytokines. Therefore, consequences of increased arginase expression should be evaluated whenever PDE inhibitors are proposed for treatment of inflammatory disorders in which IL-4 and/or TGF-beta predominate. PMID- 16257999 TI - GEF-H1 is involved in agonist-induced human pulmonary endothelial barrier dysfunction. AB - Endothelial cell (EC) permeability is precisely controlled by cytoskeletal elements [actin filaments, microtubules (MT), intermediate filaments] and cell contact protein complexes (focal adhesions, adherens junctions, tight junctions). We have recently shown that the edemagenic agonist thrombin caused partial MT disassembly, which was linked to activation of small GTPase Rho, Rho-mediated actin remodeling, cell contraction, and dysfunction of lung EC barrier. GEF-H1 is an MT-associated Rho-specific guanosine nucleotide (GDP/GTP) exchange factor, which in MT-unbound state stimulates Rho activity. In this study we tested hypothesis that GEF-H1 may be a key molecule involved in Rho activation, myosin light chain phosphorylation, actin remodeling, and EC barrier dysfunction associated with partial MT disassembly. Our results show that depletion of GEF-H1 or expression of dominant negative GEF-H1 mutant significantly attenuated permeability increase, actin stress fiber formation, and increased MLC and MYPT1 phosphorylation induced by thrombin or MT-depolymerizing agent nocodazole. In contrast, expression of wild-type or activated GEF-H1 mutants dramatically enhanced thrombin and nocodazole effects on stress fiber formation and cell retraction. These results show a critical role for the GEF-H1 in the Rho activation caused by MT disassembly and suggest GEF-H1 as a key molecule involved in cross talk between MT and actin cytoskeleton in agonist-induced Rho-dependent EC barrier regulation. PMID- 16258000 TI - Partially adenosine deaminase-deficient mice develop pulmonary fibrosis in association with adenosine elevations. AB - Adenosine, a signaling nucleoside, exhibits tissue-protective and tissue destructive effects. Adenosine levels in tissues are controlled in part by the enzyme adenosine deaminase (ADA). ADA-deficient mice accumulate adenosine levels in multiple tissues, including the lung, where adenosine contributes to the development of pulmonary inflammation and chronic airway remodeling. The present study describes the development of pulmonary fibrosis in mice that have been genetically engineered to possess partial ADA enzyme activity and, thus, accumulate adenosine over a prolonged period of time. These partially ADA deficient mice live for up to 5 mo and die from apparent respiratory distress. Detailed investigations of the lung histopathology of partially ADA-deficient mice revealed progressive pulmonary fibrosis marked by an increase in the number of pulmonary myofibroblasts and an increase in collagen deposition. In addition, in regions of the distal airways that did not exhibit fibrosis, an increase in the number of large foamy macrophages and a substantial enlargement of the alveolar air spaces suggest emphysemic changes. Furthermore, important proinflammatory and profibrotic signaling pathways, including IL-13 and transforming growth factor-beta1, were activated. Increases in tissue fibrosis were also seen in the liver and kidneys of these mice. These changes occurred in association with pronounced elevations of lung adenosine concentrations and alterations in lung adenosine receptor levels, supporting the hypothesis that elevation of endogenous adenosine is a proinflammatory and profibrotic signal in this model. PMID- 16258001 TI - RNA interference for alpha-ENaC inhibits rat lung fluid absorption in vivo. AB - We used siRNA against the alpha-ENaC (epithelial Na channel) subunit to investigate ENaC involvement in lung fluid absorption in rats by the impermeable tracer technique during baseline and after beta-adrenoceptor stimulation by terbutaline. Terbutaline stimulation of lung fluid absorption increased fluid absorption by 165% in pSi-0-pretreated rat lungs (irrelevant siRNA-generating plasmid). Terbutaline failed to increase lung fluid absorption in rats given the specific alpha-ENaC siRNA-generating plasmid (pSi-4). pSi-4 pretreatment reduced baseline lung fluid absorption by approximately 30%. alpha-ENaC was undetectable in pSi-4-pretreated lungs, regardless of condition but was normal in pSi-0 pretreated lungs. We carried out a dose-response analysis where rats were given 0 200 microg/kg body wt pSi-4, and alpha-ENaC mRNA and protein expressions were analyzed. To reach IC(50) for alpha-ENaC mRNA expression, 32 microg/kg body wt pSi-4 was needed, and to reach IC(50) for alpha-ENaC protein expression, 59 microg/kg body wt pSi-4 was needed. We tested for lung tissue specificity and found no changes in beta-ENaC expression, at either mRNA or protein level, as well as no changes in alpha(1)-Na-K-ATPase protein expression. We isolated alveolar epithelial type II cells 24 h after in vivo pSi-4 pretreatment. In these cells, alpha-ENaC mRNA was undetectable, demonstrating that alveolar epithelial ENaC expression was attenuated after intratracheal alpha-ENaC siRNA-generating plasmid DNA instillation. We tested for organ specificity and found no changes in kidney alpha- and beta-ENaC mRNA and protein expression. Thus we provide conclusive evidence that beta-adrenoceptor stimulation of lung fluid absorption is critically ENaC dependent, whereas baseline lung fluid absorption seemed less ENaC dependent. PMID- 16258002 TI - Glutathione restores collagen degradation in TGF-beta-treated fibroblasts by blocking plasminogen activator inhibitor-1 expression and activating plasminogen. AB - Transforming growth factor (TGF)-beta plays an important role in tissue fibrogenesis. We previously demonstrated that reduced glutathione (GSH) supplementation blocked collagen accumulation induced by TGF-beta in NIH-3T3 cells. In the present study, we show that supplementation of GSH restores the collagen degradation rate in TGF-beta-treated NIH-3T3 cells. Restoration of collagen degradation by GSH is associated with a reduction of type I plasminogen activator inhibitor (PAI)-1 expression/activity as well as recovery of the activities of cell/extracellular matrix-associated tissue-type plasminogen activator and plasmin. Furthermore, we find that NIH-3T3 cells constitutively express plasminogen mRNA and possess plasmin activity. Blockade of cell surface binding of plasminogen/plasminogen activation with tranexamic acid (TXA) or inhibition of plasmin activity with aprotinin significantly reduces the basal level of collagen degradation both in the presence or absence of exogenous plasminogen. Most importantly, addition of TXA or active PAI-1 almost completely eliminates the restorative effects of GSH on collagen degradation in TGF-beta treated cells. Together, our results suggest that the major mechanism by which GSH restores collagen degradation in TGF-beta-treated cells is through blocking PAI-1 expression, leading to increased PA/plasmin activity and consequent proteolytic degradation of collagens. This study provides mechanistic evidence for GSH's putative therapeutic effect in the treatment of fibrotic disorders. PMID- 16258004 TI - Congenital adrenal hyperplasia: family screening using 17-hydroxyprogesterone (17 OHP) response to adrenocorticotropin (ACTH). PMID- 16258003 TI - Inhaled nitric oxide induced NOS inhibition and rebound pulmonary hypertension: a role for superoxide and peroxynitrite in the intact lamb. AB - Previous in vivo studies indicate that inhaled nitric oxide (NO) decreases nitric oxide synthase (NOS) activity and that this decrease is associated with significant increases in pulmonary vascular resistance (PVR) upon the acute withdrawal of inhaled NO (rebound pulmonary hypertension). In vitro studies suggest that superoxide and peroxynitrite production during inhaled NO therapy may mediate these effects, but in vivo data are lacking. The objective of this study was to determine the role of superoxide in the decrease in NOS activity and rebound pulmonary hypertension associated with inhaled NO therapy in vivo. In control lambs, 24 h of inhaled NO (40 ppm) decreased NOS activity by 40% (P<0.05) and increased endothelin-1 levels by 64% (P<0.05). Withdrawal of NO resulted in an acute increase in PVR (60.7%, P<0.05). Associated with these changes, superoxide and peroxynitrite levels increased more than twofold (P<0.05) following 24 h of inhaled NO therapy. However, in lambs treated with polyethylene glycol-conjugated superoxide dismutase (PEG-SOD) during inhaled NO therapy, there was no change in NOS activity, no increase in superoxide or peroxynitrite levels, and no increase in PVR upon the withdrawal of inhaled NO. In addition, endothelial NOS nitration was 18-fold higher (P<0.05) in control lambs than in PEG-SOD-treated lambs following 24 h of inhaled NO. These data suggest that superoxide and peroxynitrite participate in the decrease in NOS activity and rebound pulmonary hypertension associated with inhaled NO therapy. Reactive oxygen species scavenging may be a useful therapeutic strategy to ameliorate alterations in endogenous NO signaling during inhaled NO therapy. PMID- 16258005 TI - Impact of the MDM2 SNP309 and p53 Arg72Pro polymorphism on age of tumour onset in Li-Fraumeni syndrome. AB - Li-Fraumeni syndrome, resulting from p53 (TP53) germline mutations, represents one of the most devastating genetic predispositions to cancer. Recently, the MDM2 SNP309 (T-->G variation) was shown to be associated with accelerated tumour formation in p53 mutation carriers. The impact of the common p53 codon 72 polymorphism on cancer risk remains controversial. We therefore investigated the effect of these two polymorphisms in 61 French carriers of the p53 germline mutation. The mean age of tumour onset in MDMD2 SNP309 G allele carriers (19.6 years) was significantly different from that observed in patients homozygous for the T allele (29.9 years, p<0.05). For the p53 codon 72 polymorphism, the mean age of tumour onset in Arg allele carriers (21.8 years) was also different from that of Pro/Pro patients (34.4 years, p<0.05). We observed a cumulative effect of both polymorphisms because the mean ages of tumour onset in carriers of the MDM2G and p53Arg alleles (16.9 years) and those with the MDM2T/T and p53Pro/Pro genotypes (43 years) were clearly different (p<0.02). Therefore, our results confirm the impact of the MDM2 SNP309 G allele on the age of tumour onset in germline p53 mutation carriers, and suggest that this effect may be amplified by the p53 72Arg allele. Polymorphisms affecting p53 degradation therefore represent one of the rare examples of modifier genetic factors identified to date in mendelian predispositions to cancer. PMID- 16258006 TI - Balanced translocation in a patient with craniosynostosis disrupts the SOX6 gene and an evolutionarily conserved non-transcribed region. AB - Craniosynostosis is a congenital developmental disorder involving premature fusion of cranial sutures, which results in an abnormal shape of the skull. Significant progress in understanding the molecular basis of this phenotype has been made for a small number of syndromic craniosynostosis forms. Nevertheless, in the majority of the approximately 100 craniosynostosis syndromes and in non syndromic craniosynostosis the underlying gene defects and pathomechanisms are unknown. Here we report on a male infant presenting at birth with brachycephaly, proptosis, midfacial hypoplasia, and low set ears. Three dimensional cranial computer tomography showed fusion of the lambdoid sutures and distal part of the sagittal suture with a gaping anterior fontanelle. Mutations in the genes for FGFR2 and FGFR3 were excluded. Standard chromosome analysis revealed a de novo balanced translocation t(9;11)(q33;p15). The breakpoint on chromosome 11p15 disrupts the SOX6 gene, known to be involved in skeletal growth and differentiation processes. SOX6 mutation screening of another 104 craniosynostosis patients revealed one missense mutation leading to the exchange of a highly conserved amino acid (p.D68N) in a single patient and his reportedly healthy mother. The breakpoint on chromosome 9 is located in a region without any known or predicted genes but, interestingly, disrupts patches of evolutionarily highly conserved non-genic sequences and may thus led to dysregulation of flanking genes on chromosome 9 or 11 involved in skull vault development. The present case is one of the very rare reports of an apparently balanced translocation in a patient with syndromic craniosynostosis, and reveals novel candidate genes for craniosynostoses and cranial suture formation. PMID- 16258007 TI - An empirical comparison of case-control and trio based study designs in high throughput association mapping. AB - Motivated by high throughput genotyping technology, our aim in this study was to experimentally compare the power and accuracy of case-control and family trio based approaches for haplotype based, large scale, association gene mapping. We compared trio based and case-control study designs in different disease models, and partitioned the performance differences into separate components: those from the sample ascertainment, the effective sample size, and the haplotyping approaches. For systematic and controlled tests, we simulated a rapidly expanding and relatively young isolated population. The experiments were also replicated with real asthma data. We used computationally efficient methods that scale up to large amounts of both markers and individuals. Mapping is based on a haplotype association test for haplotypes of 1-10 markers. For population based haplotype reconstruction, we use HaploRec, and compare it to both a simple trio based inference and true haplotypes. Firstly and surprisingly, statistically inferred population based haplotypes can be equally powerful as true haplotypes. Secondly, as expected, the effective sample size has a clear effect on both gene detection power and mapping accuracy. Thirdly, the sample ascertainment method does not have much effect on mapping accuracy. Finally, an interesting side result is that the simple haplotype association test clearly outperformed exhaustive allelic transmission disequilibrium tests. The results suggest that the case-control design is a powerful alternative to the more laborious family based ascertainment approach, especially for large datasets, and wherever population stratification can be controlled. PMID- 16258008 TI - Cloning and molecular characterization of the basic peroxidase isoenzyme from Zinnia elegans, an enzyme involved in lignin biosynthesis. AB - The major basic peroxidase from Zinnia elegans (ZePrx) suspension cell cultures was purified and cloned, and its properties and organ expression were characterized. The ZePrx was composed of two isoforms with a M(r) (determined by matrix-assisted laser-desorption ionization time of flight) of 34,700 (ZePrx34.70) and a M(r) of 33,440 (ZePrx33.44). Both isoforms showed absorption maxima at 403 (Soret band), 500, and 640 nm, suggesting that both are high-spin ferric secretory class III peroxidases. M(r) differences between them were due to the glycan moieties, and were confirmed from the total similarity of the N terminal sequences (LSTTFYDTT) and by the 99.9% similarity of the tryptic fragment fingerprints obtained by reverse-phase nano-liquid chromatography. Four full-length cDNAs coding for these peroxidases were cloned. They only differ in the 5'-untranslated region. These differences probably indicate different ways in mRNA transport, stability, and regulation. According to the k(cat) and apparent K(m)(RH) values shown by both peroxidases for the three monolignols, sinapyl alcohol was the best substrate, the endwise polymerization of sinapyl alcohol by both ZePrxs yielding highly polymerized lignins with polymerization degrees > or =87. Western blots using anti-ZePrx34.70 IgGs showed that ZePrx33.44 was expressed in tracheary elements, roots, and hypocotyls, while ZePrx34.70 was only expressed in roots and young hypocotyls. None of the ZePrx isoforms was significantly expressed in either leaves or cotyledons. A neighbor-joining tree constructed for the four full-length cDNAs suggests that the four putative paralogous genes encoding the four cDNAs result from duplication of a previously duplicated ancestral gene, as may be deduced from the conserved nature and conserved position of the introns. PMID- 16258009 TI - Reconstitution and properties of the recombinant glyceraldehyde-3-phosphate dehydrogenase/CP12/phosphoribulokinase supramolecular complex of Arabidopsis. AB - Calvin cycle enzymes glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and phosphoribulokinase (PRK) form together with the regulatory peptide CP12 a supramolecular complex in Arabidopsis (Arabidopsis thaliana) that could be reconstituted in vitro using purified recombinant proteins. Both enzyme activities were strongly influenced by complex formation, providing an effective means for regulation of the Calvin cycle in vivo. PRK and CP12, but not GapA (A(4) isoform of GAPDH), are redox-sensitive proteins. PRK was reversibly inhibited by oxidation. CP12 has no enzymatic activity, but it changed conformation depending on redox conditions. GapA, a bispecific NAD(P)-dependent dehydrogenase, specifically formed a binary complex with oxidized CP12 when bound to NAD. PRK did not interact with either GapA or CP12 singly, but oxidized PRK could form with GapA/CP12 a stable ternary complex of about 640 kD (GapA/CP12/PRK). Exchanging NADP for NAD, reducing CP12, or reducing PRK were all conditions that prevented formation of the complex. Although GapA activity was little affected by CP12 alone, the NADPH-dependent activity of GapA embedded in the GapA/CP12/PRK complex was 80% inhibited in respect to the free enzyme. The NADH activity was unaffected. Upon binding to GapA/CP12, the activity of oxidized PRK dropped from 25% down to 2% the activity of the free reduced enzyme. The supramolecular complex was dissociated by reduced thioredoxins, NADP, 1,3 bisphosphoglycerate (BPGA), or ATP. The activity of GapA was only partially recovered after complex dissociation by thioredoxins, NADP, or ATP, and full GapA activation required BPGA. NADP, ATP, or BPGA partially activated PRK, but full recovery of PRK activity required thioredoxins. The reversible formation of the GapA/CP12/PRK supramolecular complex provides novel possibilities to finely regulate GapA ("non-regulatory" GAPDH isozyme) and PRK (thioredoxin sensitive) in a coordinated manner. PMID- 16258010 TI - An egg apparatus-specific enhancer of Arabidopsis, identified by enhancer detection. AB - Despite a central role in angiosperm reproduction, few gametophyte-specific genes and promoters have been isolated, particularly for the inaccessible female gametophyte (embryo sac). Using the Ds-based enhancer-detector line ET253, we have cloned an egg apparatus-specific enhancer (EASE) from Arabidopsis (Arabidopsis thaliana). The genomic region flanking the Ds insertion site was further analyzed by examining its capability to control gusA and GFP reporter gene expression in the embryo sac in a transgenic context. Through analysis of a 5' and 3' deletion series in transgenic Arabidopsis, the sequence responsible for egg apparatus-specific expression was delineated to 77 bp. Our data showed that this enhancer is unique in the Arabidopsis genome, is conserved among different accessions, and shows an unusual pattern of sequence variation. This EASE works independently of position and orientation in Arabidopsis but is probably not associated with any nearby gene, suggesting either that it acts over a large distance or that a cryptic element was detected. Embryo-specific ablation in Arabidopsis was achieved by transactivation of a diphtheria toxin gene under the control of the EASE. The potential application of the EASE element and similar control elements as part of an open-source biotechnology toolkit for apomixis is discussed. PMID- 16258011 TI - The cold-induced early activation of phospholipase C and D pathways determines the response of two distinct clusters of genes in Arabidopsis cell suspensions. AB - In plants, a temperature downshift represents a major stress that will lead to the induction or repression of many genes. Therefore, the cold signal has to be perceived and transmitted to the nucleus. In response to a cold exposure, we have shown that the phospholipase D (PLD) and the phospholipase C (PLC)/diacylglycerol kinase pathways are simultaneously activated. The role of these pathways in the cold response has been investigated by analyzing the transcriptome of cold treated Arabidopsis (Arabidopsis thaliana) suspension cells in the presence of U73122 or ethanol, inhibitors of the PLC/diacylglycerol kinase pathway and of the phosphatidic acid produced by PLD, respectively. This approach showed that the expression of many genes was modified by the cold response in the presence of such agents. The cold responses of most of the genes were repressed, thus correlating with the inhibitory effect of U73122 or ethanol. We were thus able to identify 58 genes that were regulated by temperature downshift via PLC activity and 87 genes regulated by temperature downshift via PLD-produced phosphatidic acid. Interestingly, each inhibitor appeared to affect different cold response genes. These results support the idea that both the PLC and PLD pathways are upstream of two different signaling pathways that lead to the activation of the cold response. The connection of these pathways with the CBF pathway, currently the most understood genetic system playing a role in cold acclimation, is discussed. PMID- 16258012 TI - Transcriptome analysis reveals specific modulation of abscisic acid signaling by ROP10 small GTPase in Arabidopsis. AB - Abscisic acid (ABA) is a hormone that modulates a variety of agronomically important growth and developmental processes and various stresses responses, but its signal transduction pathways remain poorly understood. ROP10, a member of ROP small GTPases in Arabidopsis (Arabidopsis thaliana), is a plasma membrane associated protein specifically involved in negative regulation of ABA responses. To dissect the ROP10-mediated ABA signaling, we carried out transcriptome analysis using the Arabidopsis full-genome chip. Our analysis revealed a total of 262 and 125 genes that were, respectively, up- and down-regulated (> or =2-fold cutoff) by 1 mum ABA in wild type (Wassilewskija [Ws]); 42 up-regulated and 38 down-regulated genes have not been identified in other studies. Consistent with the nonpleiotropic phenotypes of rop10-1, only three genes were altered in rop10 1 in the absence of ABA treatment. In response to 1 microm ABA, 341 and 127 genes were, respectively, activated and repressed in rop10-1. Interestingly, a particular subset of 21 genes that were not altered by 1 microm ABA in Ws but only activated in rop10-1 was identified. Reverse transcription-polymerase chain reaction analysis revealed the existence of three distinct categories of ABA dose response patterns. One novel category is characterized by their ABA unresponsiveness in Ws and activation in rop10-1 at 1 microm but not 10 and 100 microm of ABA. This indicates that ROP10 gates the expression of genes that are specific to low concentrations of ABA. Furthermore, almost all of these 21 genes are known to be highly induced by various biotic and abiotic stresses. Consequently, we found that rop10-1 enhanced the sensitivity of seed germination inhibition to mannitol and sodium chloride. Our results suggest that ROP10 negatively regulates ABA responses by specifically and differentially modulating the ABA sensitivity of a subset of genes including protein kinases and zinc finger family proteins. PMID- 16258013 TI - The rib1 mutant of Arabidopsis has alterations in indole-3-butyric acid transport, hypocotyl elongation, and root architecture. AB - Polar transport of the auxin indole-3-butyric acid (IBA) has recently been shown to occur in Arabidopsis (Arabidopis thaliana) seedlings, yet the physiological importance of this process has yet to be fully resolved. Here we describe the first demonstration of altered IBA transport in an Arabidopsis mutant, and show that the resistant to IBA (rib1) mutation results in alterations in growth, development, and response to exogenous auxin consistent with an important physiological role for IBA transport. Both hypocotyl and root IBA basipetal transport are decreased in rib1 and root acropetal IBA transport is increased. While indole-3-acetic acid (IAA) transport levels are not different in rib1 compared to wild type, root acropetal IAA transport is insensitive to the IAA efflux inhibitor naphthylphthalamic acid in rib1, as is the dependent physiological process of lateral root formation. These observed changes in IBA transport are accompanied by altered rib1 phenotypes. Previously, rib1 roots were shown to be less sensitive to growth inhibition by IBA, but to have a wild-type response to IAA in root elongation. rib1 is also less sensitive to IBA in stimulation of lateral root formation and in hypocotyl elongation under most, but not all, light and sucrose conditions. rib1 has wild-type responses to IAA, except under one set of conditions, low light and 1.5% sucrose, in which both hypocotyl elongation and lateral root formation show altered IAA response. Taken together, our results support a model in which endogenous IBA influences wild type seedling morphology. Modifications in IBA distribution in seedlings affect hypocotyl and root elongation, as well as lateral root formation. PMID- 16258014 TI - HKT1 mediates sodium uniport in roots. Pitfalls in the expression of HKT1 in yeast. AB - The function of HKT1 in roots is controversial. We tackled this controversy by studying Na+ uptake in barley (Hordeum vulgare) roots, cloning the HvHKT1 gene, and expressing the HvHKT1 cDNA in yeast (Saccharomyces cerevisiae) cells. High affinity Na+ uptake was not detected in plants growing at high K+ but appeared soon after exposing the plants to a K(+)-free medium. It was a uniport, insensitive to external K+ at the beginning of K+ starvation and inhibitable by K+ several hours later. The expression of HvHKT1 in yeast was Na+ (or K+) uniport, Na(+)-K+ symport, or a mix of both, depending on the construct from which the transporter was expressed. The Na+ uniport function was insensitive to external K+ and mimicked the Na+ uptake carried out by the roots at the beginning of K+ starvation. The K+ uniport function only took place in yeast cells that were completely K+ starved and disappeared when internal K+ increased, which makes it unlikely that HvHKT1 mediates K+ uptake in roots. Mutation of the first in-frame AUG codon of HvHKT1 to CUC changed the uniport function into symport. The expression of the symport from either mutants or constructs keeping the first in-frame AUG took place only in K(+)-starved cells, while the uniport was expressed in all conditions. We discuss here that the symport occurs only in heterologous expression. It is most likely related to the K+ inhibitable Na+ uptake process of roots that heterologous systems fail to reproduce. PMID- 16258015 TI - Variations in CYP74B2 (hydroperoxide lyase) gene expression differentially affect hexenal signaling in the Columbia and Landsberg erecta ecotypes of Arabidopsis. AB - The CYP74B2 gene in Arabidopsis (Arabidopsis thaliana) ecotype Columbia (Col) contains a 10-nucleotide deletion in its first exon that causes it to code for a truncated protein not containing the P450 signature typical of other CYP74B subfamily members. Compared to CYP74B2 transcripts in the Landsberg erecta (Ler) ecotype that code for full-length hydroperoxide lyase (HPL) protein, CYP74B2 transcripts in the Col ecotype accumulate at substantially reduced levels. Consistent with the nonfunctional HPL open reading frame in the Col ecotype, in vitro HPL activity analyses using either linoleic acid hydroperoxide or linolenic acid hydroperoxide as substrates show undetectable HPL activity in the Col ecotype and C6 volatile analyses using leaf homogenates show substantially reduced amounts of hexanal and no detectable trans-2-hexenal generated in the Col ecotype. P450-specific microarrays and full-genome oligoarrays have been used to identify the range of other transcripts expressed at different levels in these two ecotypes potentially as a result of these variations in HPL activity. Among the transcripts expressed at significantly lower levels in Col leaves are those coding for enzymes involved in the synthesis of C6 volatiles (LOX2, LOX3), jasmonates (OPR3, AOC), and aliphatic glucosinolates (CYP83A1, CYP79F1, AOP3). Two of the three transcripts coding for aliphatic glucosinolates (CYP83A1, AOP3) are also expressed at significantly lower levels in Col flowers. PMID- 16258016 TI - Independent roles for EARLY FLOWERING 3 and ZEITLUPE in the control of circadian timing, hypocotyl length, and flowering time. AB - The circadian clock regulates many aspects of plant development, including hypocotyl elongation and photoperiodic induction of flowering. ZEITLUPE (ZTL) is a clock-related F-box protein, and altered ZTL expression causes fluence rate dependent circadian period effects, and altered hypocotyl elongation and flowering time. EARLY FLOWERING 3 (ELF3) is a novel protein of unknown biochemical function. elf3 mutations cause light-dependent circadian dysfunction, elongated hypocotyls, and early flowering. Although both genes affect similar processes, their relationship is unclear. Here we show that the effects of ZTL and ELF3 on circadian clock function and early photomorphogenesis are additive. The long period of ztl mutations and ELF3 overexpressors are more severe than either alone. Dark-release experiments showing additivity in phase advances suggest that the arrthymicity caused by ZTL overexpression and that of the elf3-1 mutation arise through independent pathways. A similar additive effect on hypocotyl elongation in red and blue light is also observed. In contrast, ELF3 and ZTL overexpressors act similarly to control flowering time in long days through the CONSTANS/FLOWERING LOCUS T (CO/FT) pathway. ZTL overexpression does not delay flowering through changes in GIGANTEA or FLAVIN-BINDING, KELCH REPEAT, F-BOX levels, but through a ZTL-mediated reduction in CO expression. In contrast, ELF3 negatively regulates CO, FT, and GIGANTEA transcript levels, as the expression of all three genes is increased in elf3-1. The elf3-1 co-1 double mutant flowers much earlier in long days than co-1, although FT message levels remain very low. These results show that elf3-1 can derepress late flowering through a CO-independent mechanism. ELF3 may act at more than one juncture, possibly posttranscriptionally. PMID- 16258017 TI - 12-oxo-phytodienoic acid triggers expression of a distinct set of genes and plays a role in wound-induced gene expression in Arabidopsis. AB - Jasmonic acid (JA) and methyl jasmonate (MeJA), collectively known as JAs, regulate diverse physiological processes in plants, including the response to wounding. Recent reports suggest that a cyclopentenone precursor of JA, 12-oxo phytodienoic acid (OPDA), can also induce gene expression. However, little is known about the physiological significance of OPDA-dependent gene expression. We used microarray analysis of approximately 21,500 Arabidopsis (Arabidopsis thaliana) genes to compare responses to JA, MeJA, and OPDA treatment. Although many genes responded identically to both OPDA and JAs, we identified a set of genes (OPDA-specific response genes [ORGs]) that specifically responded to OPDA but not to JAs. ORGs primarily encoded signaling components, transcription factors, and stress response-related genes. One-half of the ORGs were induced by wounding. Analysis using mutants deficient in the biosynthesis of JAs revealed that OPDA functions as a signaling molecule in the wounding response. Unlike signaling via JAs, OPDA signaling was CORONATINE INSENSITIVE 1 independent. These results indicate that an OPDA signaling pathway functions independently of JA/MeJA signaling and is required for the wounding response in Arabidopsis. PMID- 16258020 TI - Differential in vivo potencies of naltrexone and 6beta-naltrexol in the monkey. AB - 6beta-Naltrexol is the major metabolite of the opioid receptor antagonist, naltrexone, in humans. However, there are no functional studies of 6beta naltrexol in primates. The aim of this study was to compare the in vitro and in vivo potencies of naltrexone and 6beta-naltrexol in rhesus monkeys. Affinity and potency were determined using radioligand displacement and stimulation of 5'-O-(3 [(35)S]thio)triphosphate ([(35)S]GTPgammaS) binding in monkey brain membranes. In vivo apparent pA(2) analysis was applied to compare the mu-opioid receptor (MOR) antagonist potency of both compounds in nondependent monkeys. In addition, the potencies of both compounds were determined in precipitating withdrawal manifested by increased respiratory parameters in acute morphine-dependent monkeys. In vitro assays revealed that naltrexone displayed 2-fold higher affinity and potency than 6beta-naltrexol for the MOR binding site and for MOR agonist-stimulated [(35)S]GTPgammaS binding, respectively. 6beta-Naltrexol (0.32 3.2 mg/kg) dose-dependently produced parallel rightward shifts of the dose response curve of alfentanil-induced antinociception. Nevertheless, the apparent pA(2) value of 6beta-naltrexol (6.5) was 100-fold less potent than that of naltrexone (8.5) determined previously. 6beta-Naltrexol was also less potent than naltrexone in antagonizing other MOR-mediated effects including respiratory depression and itch/scratching. Naltrexone (0.0032-0.032 mg/kg) and 6beta naltrexol (0.32-3.2 mg/kg) retained the same potency difference in precipitating withdrawal to a similar degree. Furthermore, 6beta-naltrexol failed to block naltrexone-precipitated withdrawal in morphine-dependent monkeys. These results indicate that naltrexone and 6beta-naltrexol display similar pharmacological actions with a large in vivo potency difference in monkeys such that 6beta naltrexol may play a minimal role in the therapeutic or antagonist effects of naltrexone in primates. PMID- 16258019 TI - Arabidopsis vegetative storage protein is an anti-insect acid phosphatase. AB - Indirect evidence previously suggested that Arabidopsis (Arabidopsis thaliana) vegetative storage protein (VSP) could play a role in defense against herbivorous insects. To test this hypothesis, other AtVSP-like sequences in Arabidopsis were identified through a Basic Local Alignment Search Tool search, and their transcriptional profiles were investigated. In response to methyl jasmonate application or phosphate starvation, AtVSP and AtVSP-like genes exhibited differential expression patterns, suggesting distinct roles played by each member. Arabidopsis VSP2 (AtVSP2), a gene induced by wounding, methyl jasmonate, insect feeding, and phosphate deprivation, was selected for bacterial expression and functional characterization. The recombinant protein exhibited a divalent cation-dependent phosphatase activity in the acid pH range. When incorporated into the diets of three coleopteran and dipteran insects that have acidic gut lumen, recombinant AtVSP2 significantly delayed development of the insects and increased their mortality. To further determine the biochemical basis of the anti insect activity of the protein, the nucleophilic aspartic acid-119 residue at the conserved DXDXT signature motif was substituted by glutamic acid via site directed mutagenesis. This single-amino acid alteration did not compromise the protein's secondary or tertiary structure, but resulted in complete loss of its acid phosphatase activity as well as its anti-insect activity. Collectively, we conclude that AtVSP2 is an anti-insect protein and that its defense function is correlated with its acid phosphatase activity. PMID- 16258018 TI - Gibberellins are involved in nodulation of Sesbania rostrata. AB - Upon submergence, Azorhizobium caulinodans infects the semiaquatic legume Sesbania rostrata via the intercellular crack entry process, resulting in lateral root-based nodules. A gene encoding a gibberellin (GA) 20-oxidase, SrGA20ox1, involved in GA biosynthesis, was transiently up-regulated during lateral root base nodulation. Two SrGA20ox1 expression patterns were identified, one related to intercellular infection and a second observed in nodule meristem descendants. The infection-related expression pattern depended on bacterially produced nodulation (Nod) factors. Pharmacological studies demonstrated that GAs were involved in infection pocket and infection thread formation, two Nod factor dependent events that initiate lateral root base nodulation, and that they were also needed for nodule primordium development. Moreover, GAs inhibited the root hair curling process. These results show that GAs are Nod factor downstream signals for nodulation in hydroponic growth. PMID- 16258021 TI - A novel cannabinoid peripheral cannabinoid receptor-selective inverse agonist blocks leukocyte recruitment in vivo. AB - The expression of the cannabinoid peripheral cannabinoid receptor (CB(2)) receptor on peripheral immune cells suggests that compounds specific for CB(2) might be effective anti-inflammatory agents. In this report, we present the initial biological profiling of a novel triaryl bis-sulfone, Sch.336 (N-[1(S)-[4 [[4-methoxy-2-[(4-methoxyphenyl)sulfonyl]phenyl] sulfonyl]phenyl]ethyl]methanesulfonamide), which is selective for the human cannabinoid CB(2) receptor (hCB(2)). Sch.336 is an inverse agonist at hCB(2), as shown by its ability to decrease guanosine 5'-3-O-(thio)triphosphate (GTPgammaS) binding to membranes containing hCB(2), by the ability of GTPgammaS to left-shift Sch.336 binding to hCB(2) in these membranes, and by the compound's ability to increase forskolin-stimulated cAMP levels in CHO cells expressing hCB(2). In these systems, Sch.336 displays a greater potency than that reported for the CB(2)-selective dihydropyrazole, SR144528 (N-[(1S)-endo-1,3,3-trimethylbicyclo [2.2.1]heptan2-yl]-5-(4-chloro-3-methylphenyl)-1-[(4-methylphenyl)methyl]-1H pyrazole-3-carboxamide). In vitro, Sch.336 impairs the migration of CB(2) expressing recombinant cell lines to the cannabinoid agonist 2 arachidonylglycerol. In vivo, the compound impairs migration of cells to cannabinoid agonist HU210 [(6aR)-trans-3-(1,1-dimethylheptyl)-6a,7,10,10a tetrahydro-1-hydroxy-6,6-dimethyl-6H-dibenzo [b,d] pyran-9-methanol]. Oral administration of the Sch.336 significantly inhibited leukocyte trafficking in several rodent in vivo models, induced either by specific chemokines or by antigen challenge. Finally, oral administration of Sch.336 blocked ovalbumin induced lung eosinophilia in mice, a disease model for allergic asthma. We conclude that selective cannabinoid CB(2) inverse agonists may serve as novel immunomodulatory agents in the treatment of a broad range of acute and chronic inflammatory disorders in which leukocyte recruitment is a hallmark of disease pathology. PMID- 16258022 TI - DNA repair enzyme, O6-methylguanine DNA methyltransferase, modulates cytotoxicity of camptothecin-derived topoisomerase I inhibitors. AB - Two camptothecin-resistant cell lines, CPT30 and KB100, were established and characterized previously in our laboratory. Because enhanced sensitivity to 1,3 bis(2-chloroethyl)-1-nitrosourea (BCNU) and decreased expression of O(6) methylguanine-DNA methyltransferase (MGMT) protein were observed in these lines, we hypothesized that MGMT may be a determinant of cytotoxicity associated with camptothecin-derived DNA topoisomerase I inhibitors (CPTs). We used the Tet-On system to induce expression of MGMT in Chinese hamster ovary (CHO) cells and RNA interference to knock down MGMT expression in human nasopharyngeal carcinoma HONE 1 cells in order to identify any correlations between MGMT expression and CPTs cytotoxicity. CHO-derived Tet-On-inducible cells (S12+) showed MGMT overexpression and statistically significant more resistance to BCNU, camptothecin, 7-ethyl-10-hydrocamptothecin (SN38), and topotecan than parental CHO cells (p < 0.05), but there was less resistance to CPTs than to BCNU. Knockdown of MGMT expression with small interfering RNA in HONE-1 cells conferred increased sensitivity to BCNU and CPTs compared with mock control. Furthermore, alteration of MGMT expression coincides with CPT-induced cell death and poly(ADP ribose) polymerase cleavage. There were no differences in protein levels and catalytic activity of topoisomerase I between MGMT-proficient and MGMT-deficient cells from the Tet-On-inducible and small interfering RNA (siRNA) systems. Resistance to CPTs coincided with decreased amounts of protein-linked DNA breaks generated by CPTs in MGMT-proficient cells and vice versa in MGMT-deficient cells. Our data indicate that MGMT can modulate cytotoxicity of CPT-derived topoisomerase I inhibitors. PMID- 16258024 TI - Multiple transporters affect the disposition of atorvastatin and its two active hydroxy metabolites: application of in vitro and ex situ systems. AB - Atorvastatin (ATV) is primarily metabolized by CYP3A in the liver to form two active hydroxy metabolites. Therefore, the sequential transport system governed by hepatic uptake and efflux transporters is important for the drug disposition and metabolism. Here, we assessed the interaction of ATV with hepatic uptake transporter organic anion transporting polypeptide (Oatp) and efflux transporter multidrug resistance associated protein 2 (MRP2/Mrp2) in vitro and ex situ using the isolated perfused rat liver (IPRL). Rifampicin (RIF) was chosen as an inhibitor for Oatp in both uptake and IPRL studies. Its inhibitory effects on MRP2 and metabolism were also tested using MRP2-overexpressing cells and rat microsomes, respectively. Our results indicate that RIF effectively inhibits the Oatp-mediated uptake of ATV and its metabolites. Inhibition on MRP2-mediated efflux of ATV was also observed at a high RIF concentration. Compared with ATV alone in the IPRL, the area under the curve(s) (AUC) of ATV was significantly increased by RIF, whereas the AUC of both metabolites were also increased in a concentration-dependent manner. However, the extent of metabolism was significantly reduced, as reflected by the reduced amounts of metabolites detected in RIF-treated livers. In conclusion, inhibition of Oatp-mediated uptake seems to be the major determinant for interaction between ATV and RIF. Metabolites of ATV were subject to Oatp-mediated uptake as well, suggesting that they undergo a similar disposition pathway as the parent drug. These data emphasize the relevance of uptake transporter as being one of the major players in hepatic drug elimination, even for substrates that undergo metabolism. PMID- 16258023 TI - Genetic variants of the human dipeptide transporter PEPT1. AB - We tested whether genetic polymorphisms affect activity of the dipeptide transporter PEPT1, which mediates bioavailability of peptidomimetic drugs. All 23 exons and adjoining intronic sections of PEPT1 (SLC15A1) were sequenced in 247 individuals of various ethnic origins (Coriell collection). Of 38 single nucleotide polymorphisms (SNPs), 21 occurred in intronic and noncoding regions and 17 in exonic coding region, of which nine were nonsynonymous. Eight nonsynonymous variants were cloned into expression vectors and functionally characterized after transient transfection into Cos7 and Chinese hamster ovary cells. None of the variants had altered transport activity for various ligands, supporting previous results, except for the new, low-frequency PEPT1-F28Y. This variant displayed significantly reduced cephalexin uptake attributable to increased K(m). Altered pH dependence of substrate transport suggested a role for F28Y in H(+)-driven translocation. Haplotype analysis revealed significant differences among ethnic populations. To search for cis-acting polymorphisms affecting transcription and mRNA processing, we measured allelic PEPT1 mRNA expression in human intestinal biopsy samples using a frequent-marker SNP in exon 17. Of 24 heterozygous samples, significant differences in allelic mRNA levels of 20 to 30% were observed in seven tissues. However, the small difference suggests that cis-acting regulatory factors have only limited effects on transporter activity. We also measured the relative formation of a splice variant (PEPT1-RF). PEPT1-RF mRNA levels ranged from 2 to 44% of total PEPT1-related mRNA, with potential consequences for drug absorption. Together with previous results, this study reveals a relatively low level of genetic variability in polymorphisms affecting both protein function and gene regulation. PMID- 16258025 TI - On a functional equation model of transient cell growth. AB - A cell-growth model with applications to modelling the size distribution of diatoms is examined. The analytic solution to the model without dispersion is found and is shown to display periodic exponential growth rather than asynchronous (or balanced) exponential growth. It is shown that a bounding envelope (hull) of the solution to the model without dispersion takes the same shape as the limiting steady-size distribution to the dispersive case as dispersion tends to zero. The effect of variable growth rate on the shape of the hull is also discussed. PMID- 16258026 TI - Reverse cholesterol transport and cholesterol efflux in atherosclerosis. AB - Reverse cholesterol transport (RCT) is a pathway by which accumulated cholesterol is transported from the vessel wall to the liver for excretion, thus preventing atherosclerosis. Major constituents of RCT include acceptors such as high-density lipoprotein (HDL) and apolipoprotein A-I (apoA-I), and enzymes such as lecithin:cholesterol acyltransferase (LCAT), phospholipid transfer protein (PLTP), hepatic lipase (HL) and cholesterol ester transfer protein (CETP). A critical part of RCT is cholesterol efflux, in which accumulated cholesterol is removed from macrophages in the subintima of the vessel wall by ATP-binding membrane cassette transporter A1 (ABCA1) or by other mechanisms, including passive diffusion, scavenger receptor B1 (SR-B1), caveolins and sterol 27 hydroxylase, and collected by HDL and apoA-I. Esterified cholesterol in the HDL is then delivered to the liver for excretion. In patients with mutated ABCA1 genes, RCT and cholesterol efflux are impaired and atherosclerosis is increased. In studies with transgenic mice, disruption of ABCA1 genes can induce atherosclerosis. Levels of HDL are inversely correlated with incidences of cardiovascular disease. Supplementation with HDL or apoA-I can reverse atherosclerosis by accelerating RCT and cholesterol efflux. On the other hand, pro-inflammatory factors such as interferon-gamma (IFN-gamma), endotoxin, tumour necrosis factor-alpha (TNF-alpha) and interleukin-1 beta (IL-1beta), can be atherogenic by impairing RCT and cholesterol efflux, according to in vitro studies. RCT and cholesterol efflux play a major role in anti-atherogenesis, and modification of these processes may provide new therapeutic approaches to cardiovascular disease. Further research on new modifying factors for RCT and cholesterol efflux is warranted. PMID- 16258027 TI - Vascular endothelial-derived semaphorin 3 inhibits sympathetic axon growth. AB - Vascular sympathetic innervation is an important determinant of blood pressure and blood flow. The mechanisms that determine vascular sympathetic innervation are not well understood. Recent studies indicate that vascular endothelial cells (EC) express semaphorin 3A, a repulsive axon guidance cue. This suggests that EC would inhibit the growth of axons to blood vessels. The present study tests this hypothesis. RT-PCR and Western analyses confirmed that rat aortic vascular ECs expressed semaphorin 3A as well as other class 3 semaphorins (sema 3s). To determine the effects of EC-derived sema 3 on sympathetic axons, axon outgrowth was assessed in cultures of neonatal sympathetic ganglia grown for 72 h in the absence and presence of vascular EC. Nerve growth factor-induced axon growth in the presence of ECs was 50 +/- 4% (P < 0.05) of growth in the absence of ECs. ECs did not inhibit axon growth in the presence of an antibody that neutralized the activity of sema 3 (P > 0.05). RT-PCR and Western analyses also indicated that sema 3s were expressed in ECs of intact arteries. To assess the function of sema 3s in arteries, sympathetic ganglia were grown in the presence of arteries for 72 h, and the percentage of axons that grew toward the artery was determined: 44 +/- 4% of axons grew toward neonatal carotid arteries. Neutralization of sema 3s or removal of EC increased the percentage of axons that grew toward the artery (71 +/- 8% and 72 +/- 8%, respectively). These data indicate that vascular EC-derived sema 3s inhibit sympathetic axon growth and may thus be a determinant of vascular sympathetic innervation. PMID- 16258028 TI - P2 antagonist PPADS attenuates responses of thin fiber afferents to static contraction and tendon stretch. AB - Injection into the arterial supply of skeletal muscle of pyridoxal phosphate-6 azophenyl-2',4'-disulfonic acid (PPADS), a P2 receptor antagonist, has been shown previously to attenuate the reflex pressor responses to both static contraction and to tendon stretch. In decerebrated cats, we tested the hypothesis that PPADS attenuated the responses of groups III and IV muscle afferents to static contraction as well as to tendon stretch. We found that injection of PPADS (10 mg/kg) into the popliteal artery attenuated the responses of both group III (n = 16 cats) and group IV afferents (n = 14 cats) to static contraction. Specifically, static contraction before PPADS injection increased the discharge rate of the group III afferents from 0.1 +/- 0.05 to 1.6 +/- 0.5 impulses/s, whereas contraction after PPADS injection increased the discharge of the group III afferents from 0.2 +/- 0.1 to only 1.0 +/- 0.5 impulses/s (P < 0.05). Likewise, static contraction before PPADS injection increased the discharge rate of the group IV afferents from 0.3 +/- 0.1 to 1.0 +/- 0.3 impulses/s, whereas contraction after PPADS injection increased the discharge of the group IV afferents from 0.2 +/- 0.1 to only 0.3 +/- 0.1 impulses/s (P < 0.05). In addition, PPADS significantly attenuated the responses of group III afferents to tendon stretch but had no effect on the responses of group IV afferents. Our findings suggest that both groups III and IV afferents are responsible for evoking the purinergic component of the exercise pressor reflex, whereas only group III afferents are responsible for evoking the purinergic component of the muscle mechanoreflex that is evoked by tendon stretch. PMID- 16258030 TI - Relationship between 5-aminoimidazole-4-carboxamide-ribotide and AMP-activated protein kinase activity in the perfused mouse heart. AB - AMP-activated protein kinase (AMPK) is a cellular energy sensor whose activity responds to AMP concentration ([AMP]). An agent that activates AMPK in cells is 5 aminoimidazole-4-carboxamide-1-riboside (AICA-riboside). Phosphorylated AICA riboside or AICA-ribotide (ZMP) is an AMP analog. It is generally assumed that ZMP accumulation does not alter [AMP]. Additionally, the effect of AICA-riboside on AMPK activity of the heart is uncertain. Two hypotheses were tested in the isolated mouse heart: 1) sufficient ZMP concentration ([ZMP]) forms to increase AMPK activity, and 2) [ZMP] accumulation increases [AMP]. Perfusion of isolated mouse hearts with Krebs-Henseleit buffer containing 0.15-2 mM AICA-riboside concentration resulted in [ZMP] of 2-8 mM. ZMP accumulation reduced phosphocreatine concentration, which increased cytosolic [AMP]. In hearts with [ZMP] less than approximately 3 mM, in vivo AMPK allosteric activity effects of ZMP were observed; AMPK phosphorylation and [AMP] were not increased. With [ZMP] between 3 and 5 mM, in vitro AMPK activity and phosphorylation increased with unchanged [AMP]. This occurred in hearts perfused with 0.25 mM AICA-riboside for 48 min and 0.5 mM AICA-riboside for 24 min. The [ZMP] resulting in 50% AMPK activity (covalent phosphorylation of AMPK) was 4.1 +/- 0.6 mM. Hearts with [ZMP] >5 mM displayed increased [AMP] and AMPK activity that was not different from hearts with similar [AMP] with no [ZMP]; the half-maximal activity of AMP was 5.6 +/- 1.6 microM. Thus, in mouse hearts, AICA-riboside was metabolized to [ZMP] adequately to increase AMPK activity. Higher [ZMP] also increased cytosolic [AMP], which affects AMPK activity. PMID- 16258029 TI - Epoxyeicosatrienoic and dihydroxyeicosatrienoic acids dilate human coronary arterioles via BK(Ca) channels: implications for soluble epoxide hydrolase inhibition. AB - Epoxyeicosatrienoic acids (EETs) are metabolized by soluble epoxide hydrolase (sEH) to form dihydroxyeicosatrienoic acids (DHETs) and are putative endothelium derived hyperpolarizing factors (EDHFs). EDHFs modulate microvascular tone; however, the chemical identity of EDHF in the human coronary microcirculation is not known. We examined the capacity of EETs, DHETs, and sEH inhibition to affect vasomotor tone in isolated human coronary arterioles (HCAs). HCAs from right atrial appendages were prepared for videomicroscopy and immunohistochemistry. In vessels preconstricted with endothelin-1, three EET regioisomers (8,9-, 11,12-, and 14,15-EET) each induced a concentration-dependent dilation that was sensitive to blockade of large-conductance Ca2+-activated K+ (BK(Ca)) channels by iberiotoxin. EET-induced dilation was not altered by endothelial denudation. 8,9 , 11,12-, and 14,15-DHET also dilated HCA via activation of BK(Ca) channels. Dilation was less with 8,9- and 14,15-DHET but was similar with 11,12-DHET, compared with the corresponding EETs. Immunohistochemistry revealed prominent expression of cytochrome P-450 (CYP450) 2C8, 2C9, and 2J2, enzymes that may produce EETs, as well as sEH, in HCA. Inhibition of sEH by 1-cyclohexyl-3 dodecylurea (CDU) enhanced dilation caused by 14,15-EET but reduced dilation observed with 11,12-EET. DHET production from exogenous EETs was reduced in vessels pretreated with CDU compared with control, as measured by liquid chromatography electrospray-ionization mass spectrometry. In conclusion, EETs and DHETs dilate HCA by activating BK(Ca) channels, supporting a role for EETs/DHETs as EDHFs in the human heart. CYP450s and sEH may be endogenous sources of these compounds, and sEH inhibition has the potential to alter myocardial perfusion, depending on which EETs are produced endogenously. PMID- 16258031 TI - Homocysteine causes cerebrovascular leakage in mice. AB - Elevated plasma homocysteine (Hcy) is associated with cerebrovascular disease and activates matrix metalloproteinases (MMPs), which lead to vascular remodeling that could disrupt the blood-brain barrier. To determine whether Hcy administration can increase brain microvascular leakage secondary to activation of MMPs, we examined pial venules by intravital video microscopy through a craniotomy in anesthetized mice. Bovine serum albumin labeled with fluorescein isothiocyanate (BSA-FITC) was injected into a carotid artery to measure extravenular leakage. Hcy (30 microM/total blood volume) was injected 10 min after FITC-BSA injection. Four groups of mice were examined: 1) wild type (WT) given vehicle; 2) WT given Hcy (WT + Hcy); 3) MMP-9 gene knockout given Hcy (MMP 9-/- + Hcy); and 4) MMP-9-/- with topical application of histamine (10(-4) M) (MMP-9-/- + histamine). In the WT + Hcy mice, leakage of FITC-BSA from pial venules was significantly (P < 0.05) greater than in the other groups. There was no significant leakage of pial microvessels in MMP-9-/- + Hcy mice. Increased cerebrovascular leakage in the MMP-9-/- + histamine group showed that microvascular permeability could still increase by a mechanism independent of MMP 9. Treatment of cultured mouse microvascular endothelial cells with 30 microM Hcy resulted in significantly greater F-actin formation than in control cells without Hcy. Treatment with a broad-range MMP inhibitor (GM-6001; 1 microM) ameliorated Hcy-induced F-actin formation. These data suggest that Hcy increases microvascular permeability, in part, through MMP-9 activation. PMID- 16258033 TI - The induction of the mating program in the phytopathogen Ustilago maydis is controlled by a G1 cyclin. AB - Our understanding of how cell cycle regulation and virulence are coordinated during the induction of fungal pathogenesis is limited. In the maize smut fungus Ustilago maydis, pathogenesis and sexual development are intricately interconnected. Furthermore, the first step in the infection process is mating, and this is linked to the cell cycle. In this study, we have identified a new G1 cyclin gene from U. maydis that we have named cln1. We investigated the roles of Cln1 in growth and differentiation in U. maydis and found that although not essential for growth, its absence produces dramatic morphological defects. We provide results that are consistent with Cln1 playing a conserved role in regulating the length of G1 and cell size, but also additional morphological functions. We also present experiments indicating that the cyclin Cln1 controls sexual development in U. maydis. Overexpression of cln1 blocks sexual development, while its absence enables the cell to express sexual determinants in conditions where wild-type cells were unable to initiate this developmental program. We conclude that Cln1 contributes to negative regulation of the timing of sexual development, and we propose the existence of a negative crosstalk between mating program and vegetative growth that may help explain why these two developmental options are incompatible in U. maydis. PMID- 16258032 TI - Vitamin E protects against photoinhibition and photooxidative stress in Arabidopsis thaliana. AB - Vitamin E is considered a major antioxidant in biomembranes, but little evidence exists for this function in plants under photooxidative stress. Leaf discs of two vitamin E mutants, a tocopherol cyclase mutant (vte1) and a homogentisate phytyl transferase mutant (vte2), were exposed to high light stress at low temperature, which resulted in bleaching and lipid photodestruction. However, this was not observed in whole plants exposed to long-term high light stress, unless the stress conditions were extreme (very low temperature and very high light), suggesting compensatory mechanisms for vitamin E deficiency under physiological conditions. We identified two such mechanisms: nonphotochemical energy dissipation (NPQ) in photosystem II (PSII) and synthesis of zeaxanthin. Inhibition of NPQ in the double mutant vte1 npq4 led to a marked photoinhibition of PSII, suggesting protection of PSII by tocopherols. vte1 plants accumulated more zeaxanthin in high light than the wild type, and inhibiting zeaxanthin synthesis in the vte1 npq1 double mutant resulted in PSII photoinhibition accompanied by extensive oxidation of lipids and pigments. The single mutants npq1, npq4, vte2, and vte1 showed little sensitivity to the stress treatments. We conclude that, in cooperation with the xanthophyll cycle, vitamin E fulfills at least two different functions in chloroplasts at the two major sites of singlet oxygen production: preserving PSII from photoinactivation and protecting membrane lipids from photooxidation. PMID- 16258034 TI - Establishment of the vernalization-responsive, winter-annual habit in Arabidopsis requires a putative histone H3 methyl transferase. AB - Winter-annual accessions of Arabidopsis thaliana are often characterized by a requirement for exposure to the cold of winter to initiate flowering in the spring. The block to flowering prior to cold exposure is due to high levels of the flowering repressor FLOWERING LOCUS C (FLC). Exposure to cold promotes flowering through a process known as vernalization that epigenetically represses FLC expression. Rapid-cycling accessions typically have low levels of FLC expression and therefore do not require vernalization. A screen for mutants in which a winter-annual Arabidopsis is converted to a rapid-cycling type has identified a putative histone H3 methyl transferase that is required for FLC expression. Lesions in this methyl transferase, EARLY FLOWERING IN SHORT DAYS (EFS), result in reduced levels of histone H3 Lys 4 trimethylation in FLC chromatin. EFS is also required for expression of other genes in the FLC clade, such as MADS AFFECTING FLOWERING2 and FLOWERING LOCUS M. The requirement for EFS to permit expression of several FLC clade genes accounts for the ability of efs lesions to suppress delayed flowering due to the presence of FRIGIDA, autonomous pathway mutations, or growth in noninductive photoperiods. efs mutants exhibit pleiotropic phenotypes, indicating that the role of EFS is not limited to the regulation of flowering time. PMID- 16258036 TI - An alternative reaction pathway of F1-ATPase suggested by rotation without 80 degrees/40 degrees substeps of a sluggish mutant at low ATP. AB - F(1)-ATPase, a water-soluble portion of F(o)F(1)-ATP synthase, is a rotary motor driven by ATP hydrolysis. The central gamma-subunit rotates in the alpha(3)beta(3) cylinder by repeating four stages of rotation: ATP-binding dwell, rapid 80 degrees substep rotation, catalytic dwell, and rapid 40 degrees substep rotation. In the catalytic dwell, at least two catalytic reactions occur-cleavage of the enzyme-bound ATP and presumably release of the hydrolyzed product(s) from the enzyme-but we found that a slow ATP cleavage mutant of F(1)-ATPase from thermophilic Bacillus PS3 rotates at low ATP concentration without substeps and the catalytic dwell. Analysis indicates that in this alternative reaction pathway the two catalytic reactions occur during the preceding long ATP-binding dwell. Thus, F(1)-ATPase can operate through (at least) two competing reaction pathways, not necessarily through a simple consecutive reaction. PMID- 16258035 TI - Simultaneous measurement of water volume and pH in single cells using BCECF and fluorescence imaging microscopy. AB - Regulation and maintenance of cell water volume and intracellular pH (pHi) are vital functions that are interdependent; cell volume regulation affects, and is in turn affected by, changes in pHi. Disruption of either function underlies various pathologies. To study the interaction and kinetics of these two mechanisms, we developed and validated a quantitative fluorescence imaging microscopy method to measure simultaneous changes in pHi and volume in single cells loaded with the fluorescent probe BCECF. CWV is measured at the excitation isosbestic wavelength, whereas pHi is determined ratiometrically. The method has a time resolution of <1 s and sensitivity to osmotic changes of approximately 1%. It can be applied in real time to virtually any cell type attached to a coverslip, independently of cellular shape and geometry. Calibration procedures and algorithms developed to transform fluorescence signals into changes in cell water volume (CWV) and examples of applications are presented. PMID- 16258037 TI - Frequency modulation atomic force microscopy reveals individual intermediates associated with each unfolded I27 titin domain. AB - In this study, we apply a dynamic atomic force microscopy (AFM) technique, frequency modulation (FM) detection, to the mechanical unfolding of single titin I27 domains and make comparisons with measurements made using the AFM contact or static mode method. Static mode measurements revealed the well-known force transition occurring at 100-120 pN in the first unfolding peak, which was less clear, or more often absent, in the subsequent unfolding peaks. In contrast, some FM-AFM curves clearly resolved a force transition associated with each of the unfolding peaks irrespective of the number of observed unfolded domains. As expected for FM-AFM, the frequency shift response of the main unfolding peaks and their intermediates could only be detected when the oscillation amplitudes used were smaller than the interaction lengths being measured. It was also shown that the forces measured for the dynamical interaction of the FM-AFM technique were significantly lower than those measured using the static mode. This study highlights the potential for using dynamic AFM for investigating biological interactions, including protein unfolding and the detection of novel unfolding intermediates. PMID- 16258038 TI - A non-Gaussian distribution quantifies distances measured with fluorescence localization techniques. AB - When single-molecule fluorescence localization techniques are pushed to their lower limits in attempts to measure ever-shorter distances, measurement errors become important to understand. Here we describe the non-Gaussian distribution of measured distances that is the key to proper interpretation of distance measurements. We test it on single-molecule high-resolution colocalization data for a known distance, 10 nm, and find that it gives the correct result, whereas interpretation of the same data with a Gaussian distribution gives a result that is systematically too large. PMID- 16258039 TI - Detection of forces and displacements along the axial direction in an optical trap. AB - We present measurements of the forces on, and displacements of, an optically trapped bead along the propagation direction of the trapping laser beam (the axial direction). In a typical experimental configuration, the bead is trapped in an aqueous solution using an oil-immersion, high-numerical-aperture objective. This refractive index mismatch complicates axial calibrations due to both a shift of the trap center along the axial direction and spherical aberrations. In this work, a known DNA template was unzipped along the axial direction and its characteristic unzipping force-extension data were used to determine 1), the location of the trap center along the axial direction; 2), the axial displacement of the bead from the trap center; and 3), the axial force exerted on the bead. These axial calibrations were obtained for trap center locations up to approximately 4 microm into the aqueous solution and with axial bead displacements up to approximately 600 nm from the trap center. In particular, the axial trap stiffness decreased substantially when the trap was located further into the aqueous solution. This approach, together with conventional lateral calibrations, results in a more versatile optical trapping instrument that is accurately calibrated in all three dimensions. PMID- 16258040 TI - Characterization of the myosin-based source for second-harmonic generation from muscle sarcomeres. AB - Several biologically important protein structures give rise to strong second harmonic generation (SHG) in their native context. In addition to high-contrast optical sections of cells and tissues, SHG imaging can provide detailed structural information based on the physical constraints of the optical effect. In this study we characterize, by biochemical and optical analysis, the critical structures underlying SHG from the complex muscle sarcomere. SHG emission arises from domains of the sarcomere containing thick filaments, even within nascent sarcomeres of differentiating myocytes. SHG from isolated myofibrils is abolished by extraction of myosin, but is unaffected by removal or addition of actin filaments. Furthermore, the polarization dependence of sarcomeric SHG is not affected by either the proportion of myosin head domains or the orientation of myosin heads. By fitting SHG polarization anisotropy readings to theoretical response curves, we find an orientation for the elemental harmonophore that corresponds well to the pitch of the myosin rod alpha-helix along the thick filament axis. Taken together, these data indicate that myosin rod domains are the key structures giving SHG from striated muscle. This study should guide the interpretation of SHG contrast in images of cardiac and skeletal muscle tissue for a variety of biomedical applications. PMID- 16258041 TI - Kinetic analysis of the metal binding mechanism of Escherichia coli manganese superoxide dismutase. AB - The acquisition of a catalytic metal cofactor is an essential step in the maturation of every metalloenzyme, including manganese superoxide dismutase (MnSOD). In this study, we have taken advantage of the quenching of intrinsic protein fluorescence by bound metal ions to continuously monitor the metallation reaction of Escherichia coli MnSOD in vitro, permitting a detailed kinetic characterization of the uptake mechanism. Apo-MnSOD metallation kinetics are "gated", zero order in metal ion for both the native Mn2+ and a nonnative metal ion (Co2+) used as a spectroscopic probe to provide greater sensitivity to metal binding. Cobalt-binding time courses measured over a range of temperatures (35-50 degrees C) reveal two exponential kinetic processes (fast and slow phases) associated with metal binding. The amplitude of the fast phase increases rapidly as the temperature is raised, reflecting the fraction of Apo-MnSOD in an "open" conformation, and its temperature dependence allows thermodynamic parameters to be estimated for the "closed" to "open" conformational transition. The sensitivity of the metallated protein to exogenously added chelator decreases progressively with time, consistent with annealing of an initially formed metalloprotein complex (k anneal = 0.4 min(-1)). A domain-separation mechanism is proposed for metal uptake by apo-MnSOD. PMID- 16258042 TI - Conformational changes in single-strand DNA as a function of temperature by SANS. AB - Small-angle neutron scattering (SANS) measurements were performed on a solution of single-strand DNA, 5'-ATGCTGATGC-3', in sodium phosphate buffer solution at 10 degrees C temperature increments from 25 degrees C to 80 degrees C. Cylindrical, helical, and random coil shape models were fitted to the SANS measurements at each temperature. All the shapes exhibited an expansion in the diameter direction causing a slightly shortened pitch from 25 degrees C to 43 degrees C, an expansion in the pitch direction with a slight decrease in the diameter from 43 degrees C to 53 degrees C, and finally a dramatic increase in the pitch and diameter from 53 degrees C to 80 degrees C. Differential scanning calorimeter scans of the sequence in solution exhibited a reversible two-state transition profile with a transition temperature of 47.5 +/- 0.5 degrees C, the midpoint of the conformational changes observed in the SANS measurements, and a calorimetric transition enthalpy of 60 +/- 3 kJ mol(-1) that indicates a broad transition as is observed in the SANS measurements. A transition temperature of 47 +/- 1 degrees C was also obtained from ultraviolet optical density measurements of strand melting scans of the single-strand DNA. This transition corresponds to unstacking of the bases of the sequence and is responsible for the thermodynamic discrepancy between its binding stability to its complementary sequence determined directly at ambient temperatures and determined from extrapolated values of the melting of the duplex at high temperature. PMID- 16258045 TI - How light-induced charge transfer accelerates the receptor-state recovery of photoactive yellow protein from its signaling state. AB - Stark (electroabsorption) spectra of the M100A mutant of photoactive yellow protein reveal that the neutral, cis cofactor of the pB intermediate undergoes a strikingly large change in the static dipole moment (|Deltamu| = 19 Debye) on photon absorption. The formation of this charge-separated species, in the excited state, precedes the cis --> trans isomerization of the pB cofactor and the regeneration of pG. The large |Deltamu|, reminiscent of that produced on the excitation of pG, we propose, induces twisting of the cis cofactor as a result of translocation of negative charge, from the hydroxyl oxygen, O1, toward the C7-C8 double bond. The biological significance of this photoinduced charge transfer reaction underlies the significantly faster regeneration of pG from pB in vitro, on the absorption of blue light. PMID- 16258044 TI - Stimulation of actin polymerization by filament severing. AB - The extent and dynamics of actin polymerization in solution are calculated as functions of the filament severing rate, using a simple model of in vitro polymerization. The model is solved by both analytic theory and stochastic-growth simulation. The results show that severing essentially always enhances actin polymerization by freeing up barbed ends, if barbed-end cappers are present. Severing has much weaker effects if only pointed-end cappers are present. In the early stages of polymerization, the polymerized-actin concentration grows exponentially as a function of time. The exponential growth rate is given in terms of the severing rate, and the latter is given in terms of the maximum slope in a polymerization time course. Severing and branching are found to act synergistically. PMID- 16258043 TI - Electrostatic influence of PsaC protein binding to the PsaA/PsaB heterodimer in photosystem I. AB - The absence of the PsaC subunit in the photosystem I (PSI) complex (native PSI complex) by mutagenesis or chemical manipulation yields a PSI core (P700-F(X) core) that also lacks subunits PsaD and PsaE and the two iron-sulfur clusters F(A) and F(B), which constitute an integral part of PsaC. In this P700-F(X) core, the redox potentials (E(m)) of the two quinones A(1A/B) and the iron-sulfur cluster F(X) as well as the corresponding protonation patterns are investigated by evaluating the electrostatic energies from the solution of the linearized Poisson-Boltzmann equation. The B-side specific Asp-B558 changes its protonation state significantly upon isolating the P700-F(X) core, being mainly protonated in the native PSI complex but ionized in the P700-F(X) core. In the P700-F(X) core, E(m)(A(1A/B)) remains practically unchanged, whereas E(m)(F(X)) is upshifted by 42 mV. With these calculated E(m) values, the electron transfer rate from A(1) to F(X) in the P700-F(X) core is estimated to be slightly faster on the A(1A) side than that of the wild type, which is consistent with kinetic measurements. PMID- 16258046 TI - Improvement of biodesulfurization rate by assembling nanosorbents on the surfaces of microbial cells. AB - To improve biodesulfurization rate is a key to industrialize biodesulfurization technology. The biodesulfurization rate is partially affected by transfer rate of substrates from organic phase to microbial cell. In this study, gamma-Al2O3 nanosorbents, which had the ability to selectively adsorb dibenzothiophene (DBT) from organic phase, were assembled on the surfaces of Pseudomonas delafieldii R-8 cell, a desulfurization strain. gamma-Al2O3 nanosorbents have the ability to adsorb DBT from oil phase, and the rate of adsorption was far higher than that of biodesulfurization. Thus, DBT can be quickly transferred to the biocatalyst surface where nanosorbents were located, which quickened DBT transfer from organic phase to biocatalyst surface and resulted in the increase of biodesulfurization rate. The desulfurization rate of the cells assembled with nanosorbents was approximately twofold higher than that of original cells. The cells assembled with nanosorbents were observed by a transmission electron microscope. PMID- 16258048 TI - Sampling effects, noise, and photobleaching in temporal image correlation spectroscopy. AB - We present an extensive investigation of the accuracy and precision of temporal image correlation spectroscopy (TICS). Using simulations of laser scanning microscopy image time series, we investigate the effect of spatiotemporal sampling, particle density, noise, sampling frequency, and photobleaching of fluorophores on the recovery of transport coefficients and number densities by TICS. We show that the recovery of transport coefficients is usually limited by spatial sampling, while the measurement of accurate number densities is restricted by background noise in an image series. We also demonstrate that photobleaching of the fluorophore causes a consistent overestimation of diffusion coefficients and flow rates, and a severe underestimation of number densities. We derive a bleaching correction equation that removes both of these biases when used to fit temporal autocorrelation functions, without increasing the number of fit parameters. Finally, we image the basal membrane of a CHO cell with EGFP/alpha-actinin, using two-photon microscopy, and analyze a subregion of this series using TICS and apply the bleaching correction. We show that the photobleaching correction can be determined simply by using the average image intensities from the time series, and we use the simulations to provide good estimates of the accuracy and precision of the number density and transport coefficients measured with TICS. PMID- 16258047 TI - Kinetics of cardiac thin-filament activation probed by fluorescence polarization of rhodamine-labeled troponin C in skinned guinea pig trabeculae. AB - A genetically engineered cardiac TnC mutant labeled at Cys-84 with tetramethylrhodamine-5-iodoacetamide dihydroiodide was passively exchanged for the endogenous form in skinned guinea pig trabeculae. The extent of exchange averaged nearly 70%, quantified by protein microarray of individual trabeculae. The uniformity of its distribution was verified by confocal microscopy. Fluorescence polarization, giving probe angle and its dispersion relative to the fiber long axis, was monitored simultaneously with isometric tension. Probe angle reflects underlying cTnC orientation. In steady-state experiments, rigor cross bridges and Ca2+ with vanadate to inhibit cross-bridge formation produce a similar change in probe orientation as that observed with cycling cross-bridges (no Vi). Changes in probe angle were found at [Ca2+] well below those required to generate tension. Cross-bridges increased the Ca2+ dependence of angle change (cooperativity). Strong cross-bridge formation enhanced Ca2+ sensitivity and was required for full change in probe position. At submaximal [Ca2+], the thin filament regulatory system may act in a coordinated fashion, with the probe orientation of Ca2+-bound cTnC significantly affected by Ca2+ binding at neighboring regulatory units. The time course of the probe angle change and tension after photolytic release [Ca2+] by laser photolysis of NP-EGTA was Ca2+ sensitive and biphasic: a rapid component approximately 10 times faster than that of tension and a slower rate similar to that of tension. The fast component likely represents steps closely associated with Ca2+ binding to site II of cTnC, whereas the slow component may arise from cross-bridge feedback. These results suggest that the thin filament activation rate does not limit the tension time course in cardiac muscle. PMID- 16258049 TI - The structure of polyunsaturated lipid bilayers important for rhodopsin function: a neutron diffraction study. AB - The structure of oriented 1-stearoyl-2-docosahexaenoyl-sn-glycero-3 phosphocholine bilayers with perdeuterated stearoyl- or docosahexaenoyl hydrocarbon chains was investigated by neutron diffraction. Experiments were conducted at two different relative humidities, 66 and 86%. At both humidities we observed that the polyunsaturated docosahexaenoyl chain has a preference to reside near the lipid water interface. That leaves voids in the bilayer center that are occupied by saturated stearoyl chain segments. This uneven distribution of saturated- and polyunsaturated chain densities is likely to result in membrane elastic stress that modulates function of integral receptor proteins like rhodopsin. PMID- 16258050 TI - Phospholipid-dependent regulation of cytochrome c3-mediated electron transport across membranes. AB - Cytochrome c3 (cyt c3) can mediate electron transport across phosphatidylcholine (PC)/cardiolipin (CL) and PC/phosphatidylglycerol (PG) membranes. A two-molecule process is involved in the electron transport across PC/CL membranes in the liquid-crystalline state. In contrast, a single-molecule process dominates the electron transport across PC/CL membranes in the gel state and PC/PG membranes in the liquid-crystalline and gel states. Namely, the electron transport mechanism differs with the phospholipid composition and membrane fluidity. The rate limiting step of the two-molecule process was lateral diffusion of cyt c3 in membranes. The rate constants for the three single-molecule process cases were similar to each other. To elucidate these reaction processes, interactions between cyt c3 and phosphate groups and between cyt c3 and the glycerol backbones of phospholipid bilayers were investigated by means of 31P and 2H solid-state NMR, respectively, for CL and PC/CL membranes. The results showed that the polar headgroups of both phosphatidylcholine and CL are involved in the binding of cyt c3. Also, cyt c3 penetrates into membranes, which would induce distortion of the lipid bilayer. The molecular mechanisms underlying the single- and two-molecule processes are discussed in terms of membrane structure. PMID- 16258051 TI - Triple helical structure and stabilization of collagen-like molecules with 4(R) hydroxyproline in the Xaa position. AB - In this study, we examine the relationships between the structure and stability of five related collagen-like molecules that have hydroxyproline residues occupying positions not observed in vertebrate collagen. Two of the molecules contain valine or threonine and form stable triple helices in water. Three of the molecules contain allo-threonine (an enantiomer of threonine), serine, or alanine, and are not stable. Using molecular dynamics simulation methods, we examine possible explanations for the stability difference, including considering the possibility that differences in solvent shielding of the essential interchain hydrogen bonds may result in differences in stability. By comparing the structures of threonine- and allo-threonine-containing molecules in six polar and nonpolar solvation conditions, we find that solvent shielding is not an adequate explanation for the stability difference. A closer examination of the peptides shows that the structures of the unstable molecules are looser, having weaker intermolecular hydrogen bonds. The weakened hydrogen bonds result from extended Yaa residue Psi-angles that prevent optimal geometry. The Phi-Psi-maps of the relevant residues suggest that each residue's most favorable Psi-angle determines the corresponding collagen-like molecule's stability. Additionally, we propose that these molecules illustrate a more general feature of triple-helical structures: interchain hydrogen bonds are always longer and weaker than ideal, so they are sensitive to relatively small changes in molecular structure. This sensitivity to small changes may explain why large stability differences often result from seemingly small changes in residue sequence. PMID- 16258052 TI - Utilizing ESEEM spectroscopy to locate the position of specific regions of membrane-active peptides within model membranes. AB - Membrane-active peptides participate in many cellular processes, and therefore knowledge of their mode of interaction with phospholipids is essential for understanding their biological function. Here we present a new methodology based on electron spin-echo envelope modulation to probe, at a relatively high resolution, the location of membrane-bound lytic peptides and to study their effect on the water concentration profile of the membrane. As a first example, we determined the location of the N-terminus of two membrane-active amphipathic peptides, the 26-mer bee venom melittin and a de novo designed 15-mer D,L-amino acid amphipathic peptide (5D-L9K6C), both of which are antimicrobial and bind and act similarly on negatively charged membranes. A nitroxide spin label was introduced to the N-terminus of the peptides and measurements were performed either in H2O solutions with deuterated model membranes or in D2O solutions with nondeuterated model membranes. The lipids used were dipalmitoyl phosphatidylcholine (DPPC) and phosphatidylglycerol (PG), (DPPC/PG (7:3 w/w)), egg phosphatidylcholine (PC) and PG (PC/PG (7:3 w/w)), and phosphatidylethanolamine (PE) and PG (PE/PG, 7:3w/w). The modulation induced by the 2H nuclei was determined and compared with a series of controls that produced a reference "ruler". Actual estimated distances were obtained from a quantitative analysis of the modulation depth based on a simple model of an electron spin situated at a certain distance from the bottom of a layer with homogeneously distributed deuterium nuclei. The N-terminus of both peptides was found to be in the solvent layer in both the DPPC/PG and PC/PG membranes. For PE/PG, a further displacement into the solvent was observed. The addition of the peptides was found to change the water distribution in the membrane, making it "flatter" and increasing the penetration depth into the hydrophobic region. PMID- 16258053 TI - Dynamics of immobilized and native Escherichia coli dihydrofolate reductase by quasielastic neutron scattering. AB - The internal dynamics of native and immobilized Escherichia coli dihydrofolate reductase (DHFR) have been examined using incoherent quasielastic neutron scattering. These results reveal no difference between the high frequency vibration mean-square displacement of the native and the immobilized E. coli DHFR. However, length-scale-dependent, picosecond dynamical changes are found. On longer length scales, the dynamics are comparable for both DHFR samples. On shorter length scales, the dynamics is dominated by local jump motions over potential barriers. The residence time for the protons to stay in a potential well is tau = 7.95 +/- 1.02 ps for the native DHFR and tau = 20.36 +/- 1.80 ps for the immobilized DHFR. The average height of the potential barrier to the local motions is increased in the immobilized DHFR, and may increase the activation energy for the activity reaction, decreasing the rate as observed experimentally. These results suggest that the local motions on the picosecond timescale may act as a lubricant for those associated with DHFR activity occurring on a slower millisecond timescale. Experiments indicate a significantly slower catalytic reaction rate for the immobilized E. coli DHFR. However, the immobilization of the DHFR is on the exterior of the enzyme and essentially distal to the active site, thus this phenomenon has broad implications for the action of drugs distal to the active site. PMID- 16258054 TI - Measuring molecular elasticity by atomic force microscope cantilever fluctuations. AB - In single-molecule mechanics experiments the molecular elasticity is usually measured from the deformation in response to a controlled applied force, e.g., via an atomic force microscope cantilever. We have tested the validity of an alternative method based on a recently developed theory. The concept is to measure the change in thermal fluctuations of the cantilever tip with and without its coupling to a rigid surface via the molecule. The new method was demonstrated by its application to the elasticity measurements of L- and P-selectin complexed with P-selectin glycoprotein ligand-1 or their respective antibodies, which showed values comparable to those measured from the slope of the force-extension curve. L- and P-selectin were found to behave as nearly linear springs capable of sustaining large forces and strains without sudden unfolding. The measured spring constants of approximately 4 and approximately 1 pN/nm for L- and P-selectin, respectively, suggest that a physiological force of approximately 100 pN would result in an approximately 200% strain for the respective selectins. PMID- 16258056 TI - Weight history and risk of endometrial cancer among Chinese women. AB - BACKGROUND: Adult obesity is a well-established risk factor for endometrial cancer. However, little is known about the association of endometrial cancer risk with body size early in life and weight change during adulthood. We investigated whether women with greater early-age body size or with greater weight change during adulthood have an increased risk of endometrial cancer. METHODS: We analysed data from a population-based case-control study of endometrial cancer conducted between 1997 and 2001 in Shanghai, China. Included in this analysis were 832 endometrial cancer cases aged 30-69 years and 846 population controls. Information on weight and height history from adolescence through adulthood was obtained via structured in-person interviews. A logistic regression model was used to derive odds ratios (ORs) and 95% confidence intervals (CIs) for endometrial cancer in association with adolescent and adult adiposity, as well as adult body weight change. All ORs were adjusted for age, education, menstrual status, duration of menstruation, number of pregnancies, oral contraceptive use, and family history of cancer. RESULTS: Perceived weights and heights during puberty that were greater than average were associated with a modestly increased risk of cancer. The association for perceived weight was substantially weakened after adjustment for current body mass index (BMI). High BMI at all adult ages significantly predicted endometrial cancer risk, with recent BMI being the strongest predictor. Further analyses disclosed that weight gain during adulthood, particularly during the peri-menopausal period (age 40-50 years), was associated with a significantly elevated risk of endometrial cancer, even among currently non-obese women. Gaining >5 kg between age 40 and 50 was related to ORs of 2.3 (95% CI 1.4-3.9) for women with a BMI<25 kg/m2 and 2.0 (95% CI 1.3-3.0) for women with BMI>or=25 kg/m2. CONCLUSIONS: Adult weight gain, particularly during the peri-menopausal period, plays a significant role in the development of endometrial cancer risk. PMID- 16258055 TI - Ultrafast transient absorption studies on photosystem I reaction centers from Chlamydomonas reinhardtii. 2: mutations near the P700 reaction center chlorophylls provide new insight into the nature of the primary electron donor. AB - The energy transfer and charge separation kinetics in several core Photosystem I particles of Chlamydomonas reinhardtii with point mutations around the PA and PB reaction center chlorophylls (Chls) have been studied using ultrafast transient absorption spectroscopy in the femtosecond to nanosecond time range to characterize the influence on the early electron transfer processes. The data have been analyzed in terms of kinetic compartment models. The adequate description of the transient absorption kinetics requires three different radical pairs in the time range up to approximately 100 ps. Also a charge recombination process from the first radical pair back to the excited state is present in all the mutants, as already shown previously for the wild-type (Muller, M. G., J. Niklas, W. Lubitz, and A. R. Holzwarth. 2003. Biophys. J. 85:3899-3922; and Holzwarth, A. R., M. G. Muller, J. Niklas, and W. Lubitz. 2005. J. Phys. Chem. B. 109:5903-59115). In all mutants, the primary charge separation occurs with the same effective rate constant within the error limits as in the wild-type (>>350 ns(-1)), which implies an intrinsic rate constant of charge separation of <1 ps( 1). The rate constant of the secondary electron transfer process is slowed down by a factor of approximately 2 in the mutant B-H656C, which lacks the ligand to the central metal of Chl PB. For the mutant A-T739V, which breaks the hydrogen bond to the keto carbonyl of Chl PA, only a slight slowing down of the secondary electron transfer is observed. Finally for mutant A-W679A, which has the Trp near the PA Chl replaced, either no pronounced effect or, at best, a slight increase on the secondary electron transfer rate constants is observed. The effective charge recombination rate constant is modified in all mutants to some extent, with the strongest effect observed in mutant B-H656C. Our data strongly suggest that the Chls of the PA and PB pair, constituting what is traditionally called the "primary electron donor P700", are not oxidized in the first electron transfer process, but rather only in the secondary electron transfer step. We thus propose a new electron transfer mechanism for Photosystem I where the accessory Chl(s) function as the primary electron donor(s) and the A0 Chl(s) are the primary electron acceptor(s). This new mechanism also resolves in a straightforward manner the difficulty with the previous mechanism, where an electron would have to overcome a distance of approximately 14 A in <1 ps in a single step. If interpreted within a scheme of single-sided electron transfer, our data suggest that the B-branch is the active branch, although parallel A branch activity cannot be excluded. All the mutations do affect to a varying extent the energy difference between the reaction center excited state RC* and the first radical pair and thus affect the rate constant of charge recombination. It is interesting to note that the new mechanism proposed is in fact analogous to the electron transfer mechanism in Photosystem II, where the accessory Chl also plays the role of the primary electron donor, rather than the special Chl pair P680 (Prokhorenko, V. and A. R. Holzwarth. 2000. J. Phys. Chem. B. 104:11563 11578). PMID- 16258057 TI - Body mass index and mortality from ischaemic heart disease in a lean population: 10 year prospective study of 220,000 adult men. AB - BACKGROUND: Increased body mass index (BMI) is known to be related to ischaemic heart disease (IHD) in populations where many are overweight (BMI>or=25 kg/m2) or obese (BMI>or=30). Substantial uncertainty remains, however, about the relationship between BMI and IHD in populations with lower BMI levels. METHODS: We examined the data from a population-based, prospective cohort study of 222,000 Chinese men aged 40-79. Relative and absolute risks of death from IHD by baseline BMI were calculated, standardized for age, smoking, and other potential confounding factors. RESULTS: The mean baseline BMI was 21.7 kg/m2, and 1942 IHD deaths were recorded during 10 years of follow-up (6.5% of all such deaths). Among men without prior vascular diseases at baseline, there was a J-shaped association between BMI and IHD mortality. Above 20 kg/m2 there was a positive association of BMI with risk, with each 2 kg/m2 higher in usual BMI associated with 12% (95% CI 6-19%, 2P=0.0001) higher IHD mortality. Below this BMI range, however, the association appeared to be reversed, with risk ratios of 1.00, 1.09, and 1.15, respectively, for men with BMI 20-21.9, 18-19.9, and <18 kg/m2. The excess IHD risk observed at low BMI levels persisted after restricting analysis to never smokers or excluding the first 3 years of follow-up, and became about twice as great after allowing for blood pressure. CONCLUSIONS: Lower BMI is associated with lower IHD risk among people in the so-called normal range of BMI values (20-25 kg/m2), but below that range the association may well be reversed. PMID- 16258058 TI - Association between socioeconomic status and adiposity in urban Cameroon. AB - BACKGROUND: As the relation between socioeconomic status (SES) and obesity may depend on the stage of development of a country, this relation is assessed in adults from urban Cameroon. METHODS: A sample comprising 1530 women and 1301 men aged 25 years and above, from 1897 households in the Biyem-Assi health area in the capital of Cameroon, Yaounde, were interviewed about their household amenities, occupation, and education. Weight, height, and waist circumference were measured and subjects were classified as obese if their BMI>or=30 kg/m2 or overweight if BMI was between 25.0 and 29.9 kg/m2. Abdominal obesity was defined by a waist circumference>or=80 cm in women and>or=94 cm in men. RESULTS: Of the sample studied 33% of women and 30% of men were overweight (P<0.08), whereas 22% of women and 7% of men were obese (P<0.001). Abdominal obesity was present in 67% of women and 18% of men (P<0.001). After adjusting for age, leisure time physical activity, alcohol consumption, and tobacco smoking, the prevalence of overweight+obesity, obesity, and abdominal obesity increased with quartiles of household amenities in both genders and with occupational level in men. CONCLUSION: SES is positively associated with adiposity in urban Cameroon after adjusting for confounding factors. PMID- 16258059 TI - Isolating gene-corrected stem cells without drug selection. AB - Progress in isolating stem cells from tissues, or generating them from adult cells by nuclear transfer, encourages attempts to use stem cells from affected individuals for gene correction and autologous therapy. Current viral vectors are efficient at introducing transgenic sequences but result in random integrations. Gene targeting, in contrast, can directly correct an affected gene, or incorporate corrective sequences into a site free from undesirable side effects, but efficiency is low. Most current targeting procedures, consequently, use positive-negative selection with drugs, often requiring >/=10 days. This drug selection causes problems with stem cells that differentiate in this time or require feeder cells, because the feeders must be drug resistant and so are not eliminated by the selection. To overcome these problems, we have developed a procedure for isolating gene-corrected stem cells free from feeder cells after 3 5 days culture without drugs. The method is still positive-negative, but the positive and negative drug-resistance genes are replaced with differently colored fluorescence genes. Gene-corrected cells are isolated by FACS. We tested the method with mouse ES cells having a mutant hypoxanthine phosphoribosyltransferase (Hprt) gene and grown on feeder cells. After 5 days in culture, gene-corrected cells were obtained free from feeder cells at a "purity" of >30%, enriched >2,000 fold and with a recovery of approximately 20%. Corrected cells were also isolated singly for clonal expansion. Our FACS-based procedure should be applicable at small or large scale to stem cells that can be cultured (with feeder cells, if necessary) for >/=3 days. PMID- 16258060 TI - Two tail-anchored protein variants, differing in transmembrane domain length and intracellular sorting, interact differently with lipids. AB - C-tail-anchored (TA) proteins often require a transmembrane domain of moderate hydrophobicity to maintain their endoplasmic reticulum residence, but the suggested role of protein-lipid interactions in this phenomenon has not been established. Here, we studied the interaction of TA proteins with lipids by differential scanning calorimetry by using a model system consisting of liposomes embedding either of two forms of cytochrome b(5): the endoplasmic reticulum resident wild-type (b(5)wt) and a mutant thereof (b(5)ext), that contains five extra nonpolar amino acids in its transmembrane domain and, therefore, reaches the plasma membrane. The proteins were incorporated into liposomes of palmitoyl oleyl-phosphatidylcholine (POPC) or POPC mixed with either distearoyl phosphatidylserine (DSPS), palmitoyl-oleyl-phosphatidylserine (POPS), distearoyl phosphatidylcholine (DSPC), or C16-ceramide (CER). POPC liposomes displayed a single thermotropic transition centered at -3.4 degrees C. When present, the second lipid formed a domain within the POPC bilayer, as indicated by the appearance of an additional peak. This peak was centered at temperatures close to 0 degrees C in the case of liposomes containing 10% CER, DSPS, and POPS and at 23 degrees C in the case of DSPC, likely reflecting a higher degree of molecular packing for DSPC domains. In DSPS/POPC, POPS/POPC, or CER/POPC, but not in DSPC/POPC liposomes, the insertion of b(5)wt increased, whereas b(5)ext decreased, the relative contribution to the total enthalpy of the higher temperature, phase-separated component. These results were confirmed with fluorescence measurements by using pyrene-labeled phospholipids. The dissimilar interaction with lipids of these two differently localized TA proteins could have implications for their intracellular sorting. PMID- 16258061 TI - Intronic regulation of matrix metalloproteinase-2 revealed by in vivo transcriptional analysis in ischemia. AB - Matrix metalloproteinase-2 (MMP-2) plays an essential role in angiogenesis and arteriogenesis, two processes critical to restoration of tissue perfusion after ischemia. MMP-2 expression is increased in tissue ischemia, but the responsible mechanisms remain unknown. We studied the transcriptional activation of the MMP-2 gene in a model of hindlimb ischemia by using various MMP-2-lacZ reporter mice and chromatin immunoprecipitation. MMP-2 activity and mRNA were increased after hindlimb ischemia. Mice with targeted deletion of MMP-2 had impaired restoration of perfusion and a high incidence of limb gangrene, indicating that MMP-2 plays a critical role in ischemia-induced revascularization. Ischemia induced the expression and binding of c-Fos, c-Jun, JunB, FosB, and Fra2 to a noncanonical activating protein-1 (AP-1) site present in the MMP-2 promoter and decreased binding of the transcriptional repressor JunD. Ischemia also activated the expression and binding of p53 to an adjacent enhancer site (RE-1) and increased expression and binding of nuclear factor of activated T-cells-c2 to consensus sequences within the first intron. Deletion of either the 5' AP-1/RE-1 region of the promoter or substitution of the first intron abolished ischemia-induced MMP-2 transcription in vivo. Thus, AP-1 transcription factors and intronic activation by nuclear factor of activated T-cells-c2 act in concert to drive ischemia induced MMP-2 transcription. These findings define a critical role for MMP-2 in ischemia-induced revascularization and identify both previously uncharacterized regulatory elements within the MMP-2 gene and the cognate transcription factors required for MMP-2 activation in vivo after tissue ischemia. PMID- 16258062 TI - Nucleoid remodeling by an altered HU protein: reorganization of the transcription program. AB - Bacterial nucleoid organization is believed to have minimal influence on the global transcription program. Using an altered bacterial histone-like protein, HUalpha, we show that reorganization of the nucleoid configuration can dynamically modulate the cellular transcription pattern. The mutant protein transformed the loosely packed nucleoid into a densely condensed structure. The nucleoid compaction, coupled with increased global DNA supercoiling, generated radical changes in the morphology, physiology, and metabolism of wild-type K-12 Escherichia coli. Many constitutive housekeeping genes involved in nutrient utilization were repressed, whereas many quiescent genes associated with virulence were activated in the mutant. We propose that, as in eukaryotes, the nucleoid architecture dictates the global transcription profile and, consequently, the behavior pattern in bacteria. PMID- 16258063 TI - Genetic reduction of embryonic leukemia-inhibitory factor production rescues placentation in SOCS3-null embryos but does not prevent inflammatory disease. AB - The suppressor of cytokine-signaling (SOCS) proteins act as negative-feedback inhibitors of cytokine and growth-factor-induced signal transduction. In vivo studies have implicated SOCS3 as a negative regulator of signaling downstream of gp130, the receptor subunit shared by IL-6-like cytokines. Mice lacking SOCS3 die at midgestation because of placental failure, and SOCS3 ablation in a cell-type specific manner results in changes in the functional outcome of gp130 signaling in response to IL-6. In this study, we show that genetic reduction of leukemia inhibitory factor (LIF) production by embryo-derived tissues is sufficient to prevent the placental defect. This establishes LIF signaling as a major physiological regulator of trophoblast differentiation in vivo. Mice deficient in both SOCS3 and LIF are born in predicted numbers and appear normal at birth but exhibit failure to thrive and high neonatal mortality. Adult SOCS3-null mice on a LIF-null background succumb to a spontaneous fatal inflammatory disease characterized by neutrophilia and inflammatory-cell tissue infiltrates. The disease spectrum mimics that seen in mice with a conditional deletion of SOCS3 in hematopoietic and endothelial cells, extending the evidence for a major role for SOCS3 in the homeostatic regulation of the inflammatory response and indicates that LIF is not required for this process. PMID- 16258064 TI - Protein synthesis required for long-term memory is induced by PKC activation on days before associative learning. AB - Protein synthesis has long been known to be required for associative learning to consolidate into long-term memory. Here we demonstrate that PKC isozyme activation on days before training can induce the synthesis of proteins necessary and sufficient for subsequent long-term memory consolidation. Bryostatin (Bryo), a macrolide lactone with efficacy in subnanomolar concentrations and a potential therapeutic for Alzheimer's disease, is a potent activator of PKC, some of whose isozymes undergo prolonged activation after associative learning. Under normal conditions, two training events with paired visual and vestibular stimuli cause short-term memory of the mollusc Hermissenda that lasts approximately 7 min. However, after 4-h exposures to Bryo (0.25 ng/ml) on two preceding days, the same two training events produced long-term conditioning that lasted >1 week and that was not blocked by anisomycin (1 mug/ml). Anisomycin, however, eliminated long term memory lasting at least 1 week after nine training events. Both the nine training events alone and two Bryo exposures plus two training event regimens caused comparably increased levels of the PKC alpha-isozyme substrate calexcitin in identified type B neurons and enhanced PKC activity in the membrane fractions. Furthermore, Bryo increased overall protein synthesis in cultured mammalian neurons by up to 60% for >3 days. The specific PKC antagonist Ro-32-0432 blocked much of this Bryo-induced protein synthesis as well as the Bryo-induced enhancement of the behavioral conditioning. Thus, Bryo-induced PKC activation produces those proteins necessary and sufficient for long-term memory on days in advance of the training events themselves. PMID- 16258065 TI - Detection and quantification of mutations in the plasma of patients with colorectal tumors. AB - The early detection of cancers through analysis of circulating DNA could have a substantial impact on morbidity and mortality. To achieve this goal, it is essential to determine the number of mutant molecules present in the circulation of cancer patients and to develop methods that are sufficiently sensitive to detect these mutations. Using a modified version of a recently developed assay for this purpose, we found that patients with advanced colorectal cancers consistently contained mutant adenomatous polyposis coli (APC) DNA molecules in their plasma. The median number of APC DNA fragments in such patients was 47,800 per ml of plasma, of which 8% were mutant. Mutant APC molecules were also detected in >60% of patients with early, presumably curable colorectal cancers, at levels ranging from 0.01% to 1.7% of the total APC molecules. These results have implications for the mechanisms through which tumor DNA is released into the circulation and for diagnostic tests based on this phenomenon. PMID- 16258066 TI - Probing helix propensity of monomers within a helical oligomer. AB - A simple strategy is proposed to assess the propensity of a given monomer to follow or not follow a particular helical scheme and to study helix reversal phenomena within helical oligomers. It consists of placing a monomer having a low helix propensity between two conformationally stable helical segments. Helix reversion then occurs preferentially at the site of this monomer, leading to the formation of isomers having P (right-handed) or M (left-handed) helicities at each of the two helical segments. The proportion between the P-P/M-M and P-M isomers is indicative of the stereochemical relations between the inserted monomer and the helical frame. Thus, xylylene or carboxylic acid anhydride spacers have been introduced between two helical oligoamides of 8-amino-2 quinolinecarboxylic acid. Both these spacers presumably lack some of the structural features that confer quinoline units with a high helix propensity. Only one species is observed in solution in the case of an anhydride spacer. This species was shown by x-ray crystallography to be a racemic mixture of P-P and M-M helices. Unexpectedly, the anhydride is consistently incorporated within helical oligoamides. For the xylylene spacer, the P-P/M-M racemate and P-M meso compound are in equal proportions in chloroform, showing that this spacer does not have a propensity to adopt any helical conformation in this solvent. However, the equilibria between the various isomers are shifted in toluene, where one species largely prevails. This species was shown by x-ray crystallography to be the P-P/M M racemate. Molecular dynamics simulations are consistent with these solution data. PMID- 16258067 TI - Concurrent working memory load can reduce distraction. AB - People have difficulty performing two tasks at once. For example, maintaining items in working memory (WM) makes people more distractible. However, different types of WM load may have different effects on attentional selection depending on whether WM load overlaps with mechanisms involved in target or distractor processing. Three experiments examined the effect of concurrent WM load on Stroop tasks, a widely used measure of executive control and inhibition. Stroop interference increased when the type of WM load overlapped with the type of information required for the target task (experiment 1). In striking contrast, Stroop interference decreased when the type of WM load overlapped with distractor processing (experiment 2). Experiment 3 replicated these results in a different Stroop task. Thus, concurrent WM load does not always impair executive control; performance depends on how contents of WM and task-relevant information overlap. The results highlight how dissociable components of WM interact with perception and executive control. PMID- 16258068 TI - The ETV6-NTRK3 chimeric tyrosine kinase suppresses TGF-beta signaling by inactivating the TGF-beta type II receptor. AB - An emerging theme in cancer biology is that although some malignancies occur through the sequential acquisition of different genetic alterations, certain dominantly acting oncoproteins such as those associated with chromosomal translocations have multiple functions and do not require additional mutations for cell transformation. The ETV6-NTRK3 (EN) chimeric tyrosine kinase, a potent oncoprotein expressed in tumors derived from multiple cell lineages, functions as a constitutively active protein tyrosine kinase. Here, we show that EN suppresses TGF-beta signaling by directly binding to the type II TGF-beta receptor, thereby preventing it from interacting with the type I TGF-beta receptor. This activity requires a functional EN protein tyrosine kinase, and type II TGF-beta receptor appears to be a direct target of EN. Our findings provide evidence for a previously undescribed mechanism by which oncogenic tyrosine kinases can block TGF-beta tumor suppressor activity. PMID- 16258069 TI - Helicobacter pylori CagA induces a transition from polarized to invasive phenotypes in MDCK cells. AB - CagA is a bacterial effector protein of Helicobacter pylori that is translocated via a type IV secretion system into gastric epithelial cells. We previously described that H. pylori require CagA to disrupt the organization and assembly of apical junctions in polarized epithelial cells. In this study, we provide evidence that CagA expression is not only sufficient to disrupt the apical junctions but also perturbs epithelial differentiation. CagA-expressing cells lose apicobasal polarity and cell-cell adhesion, extend migratory pseudopodia, and degrade basement membranes, acquiring an invasive phenotype. Expression of the CagA C-terminal domain, which contains the tyrosine phosphorylated EPIYA motifs, induces pseudopodial activity but is not sufficient to induce cell migration. Conversely, the N terminus targets CagA to the cell-cell junctions. Neither domain is sufficient to disrupt cell adhesion or cell polarity, but coexpressed in trans, the N terminus determines the localization of both polypeptides. We show that CagA induces a morphogenetic program in polarized Madin-Darby canine kidney cells resembling an epithelial-to-mesenchymal transition. We propose that altered cell-cell and cell matrix interactions may serve as an early event in H. pylori-induced carcinogenesis. PMID- 16258070 TI - Telomere length and heredity: Indications of paternal inheritance. AB - Cellular telomere length is linked to replicative life span. Telomere repeats are lost in peripheral blood cells in vivo by age, and women show less telomere attrition than men. Previous reports have indicated that telomere length and chromosome-specific telomere-length patterns partly are inherited. The mode of heredity has not been clarified, but a link to the X chromosome was recently suggested. We analyzed peripheral mononuclear cells from 49 unrelated families for telomere length using a real-time PCR method. Short-term cultured Epstein Barr virus-transformed lymphoblasts from the same individuals (n = 130) were analyzed for ability to maintain telomere length and possible gender-linked inheritance. A statistically significant association between telomere lengths comparing father-son (P = 0.011, n = 20) and father-daughter (P = 0.005, n = 22) pairs was found. However, no correlation was observed between mother-daughter (P = 0.463, n = 23) or mother-son (P = 0.577, n = 18). The father-offspring correlation was highly significant (P < 0.0001), in contrast to mother-offspring (P = 0.361). Epstein-Barr virus cultures demonstrated in most cases telomere preservation inversely related to initial mononuclear cell telomere length with short telomeres displaying the most pronounced elongation. Telomere length is inherited, and evidence for a father-to-offspring heritage of this trait was obtained, whereas in vitro telomere length maintenance was found to be dependent on the initial telomere length. PMID- 16258071 TI - Shoot-applied MeJA suppresses root nodulation in Lotus japonicus. AB - To maintain a symbiotic balance, leguminous plants have a systemic regulatory system called autoregulation of nodulation (AUT). Since AUT is schematically similar to systemic resistance found in plant-pathogen interactions, we examined the effects of methyl jasmonate (MeJA) or methyl salicylate (MeSA) on nodulation in Lotus japonicus. Shoot-applied MeJA strongly suppressed nodulation in the wild type and even hypernodulation in the har1 mutant, whereas MeSA exhibited no effect. MeJA inhibited early stages of nodulation, including infection thread formation and NIN gene expression, and also suppressed lateral root formation. These findings suggest that jasmonic acid and/or its related compounds participate in AUT signaling. PMID- 16258072 TI - Characterization of mitochondrial alternative NAD(P)H dehydrogenases in Arabidopsis: intraorganelle location and expression. AB - The intramitochondrial location of putative type II NAD(P)H dehydrogenases (NDs) in Arabidopsis was investigated by measuring the ability of isolated mitochondria to take up precursor proteins generated from cDNAs using an in vitro translation system. The mature proteins of NDA1, NDA2 and NDC1 were judged to be located on the inside of the inner membrane because they were protected from protease added after the mitochondrial outer membrane had been ruptured. In contrast, NDB1, NDB2 and NDB4 were not protected from protease digestion in mitochondria with ruptured outer membranes and were deemed to be located on the outside of the inner membrane. Expression of all ND genes was measured using quantitative reverse transcription-PCR (RT-PCR) to determine transcript abundance, and compared with expression of alternative oxidase, uncoupler proteins and selected components of the oxidative phosphorylation complexes. NDA1 and NDB2 were the most prominently expressed members in a variety of tissues, and were up-regulated in the early daytime in a diurnal manner. Analysis of array data suggested that NDA1 clustered closest to the gene encoding the P-subunit of glycine decarboxylase. Taken together with the diurnal regulation of NDA1 observed here and in other studies, this suggests that NDA1 plays a role in integrating metabolic activities of chloroplasts and mitochondria. NDA2, NDB2 and Aox1a were up-regulated in a coordinated manner under various treatments, potentially forming a complete respiratory chain capable of oxidizing matrix and cytosolic NAD(P)H. NDB1 and NDC1 were down-regulated under the same conditions and may be regarded as housekeeping genes. PMID- 16258073 TI - A direct interaction between the N terminus of adenylyl cyclase AC8 and the catalytic subunit of protein phosphatase 2A. AB - Although protein scaffolding complexes compartmentalize protein kinase A (PKA) and phosphodiesterases to optimize cAMP signaling, adenylyl cyclases, the sources of cAMP, have been implicated in very few direct protein interactions. The N termini of adenylyl cyclases are highly divergent, which hints at isoform specific interactions. Indeed, the Ca(2+)-sensitive adenylyl cyclase 8 (AC8) contains a Ca(2+)/calmodulin binding site on the N terminus that is essential for stimulation of activity by the capacitative entry of Ca(2+) in the intact cell. Here, we have used the N terminus of AC8 as a bait in a yeast two-hybrid screen of a human embryonic kidney (HEK) 293 cell cDNA library and identified the catalytic subunit of the serine/threonine protein phosphatase 2A (PP2A(C)) as a binding partner. Confirming the highly specific nature of this novel interaction, glutathione-S-transferase fusion proteins containing the full-length N terminus of AC8 affinity precipitated catalytically active PP2A(C) from both HEK293 and mouse forebrain membranes-the latter a normal source of AC8. The scaffolding subunit of PP2A (PP2A(A); 65 kDa) was also precipitated by the N terminus of AC8, indicating that AC8 may occur in a complex with the PP2A core dimer. The interaction between the N terminus of AC8 and PP2A(C) was antagonized by Ca(2+)/calmodulin. However, PP2A(C) and Ca(2+)/calmodulin did not share identical binding specificities in the N terminus of AC8. PKA-mediated phosphorylation did not influence either calmodulin or PP2A(C) association with AC8. In addition, both PP2A(C) and AC8 occurred in lipid rafts. These findings are the first demonstration of an association between adenylyl cyclase and any downstream element of cAMP signaling. PMID- 16258074 TI - Chemokine stromal cell-derived factor 1alpha induces proliferation and growth hormone release in GH4C1 rat pituitary adenoma cell line through multiple intracellular signals. AB - We used GH4C1 cells as a model to study the effects of the chemokine stromal cell derived factor 1 (SDF1) in pituitary functions. In these cells, SDF1alpha induced proliferation and growth hormone secretion, suggesting a possible regulatory role for this chemokine at pituitary level. We evaluated the intracellular signaling involved in these effects: SDF1alpha increased cytosolic [Ca(2+)] and activated Pyk2, extracellular signal-regulated kinases 1 and 2 (ERK1/2), and large conductance Ca(2+)-activated K(+) channels (BK(Ca)) channels. To correlate these intracellular effectors with the proliferative and secretory effects, we inhibited their activity using BAPTA-AM (Ca(2+) chelator), 2'-amino-3' methoxyflavone (PD98059; a mitogen-activated protein kinase kinase inhibitor), salicylate (Pyk2 inhibitor), and tetraethyl ammonium (K(+) channel blocker). All of these compounds reverted SDF1alpha-induced proliferation, suggesting the involvement of multiple intracellular pathways. Conversely, only BAPTA-AM reverted growth hormone secretion. To identify a possible cross-talk and a molecular ordering among these pathways, we tested these antagonists on SDF1alpha dependent activation of ERK1/2, Pyk2, and BK(Ca) channels. From these experiments, we observed that the inhibition of [Ca(2+)](i) increase or BK(Ca) channel activity did not affect ERK1/2 activation by SDF1alpha; Pyk2 activation was purely Ca(2+)-dependent, not involving ERK1/2 or BK(Ca) channels; and BK(Ca) channel activity was antagonized by Pyk2 but not by ERK1/2 inhibitors. These data suggest that an SDF1alpha-dependent increase of [Ca(2+)](i) activates Pyk2, which in turn regulates BK(Ca) channel activity. Conversely, ERK1/2 activation is an independent phenomenon. In conclusion, we demonstrate that SDF1alpha causes both proliferation and growth hormone release from pituitary adenoma cells, suggesting that the activation of CXCR4 may represent a novel regulatory mechanism for growth hormone secretion and pituitary cell proliferation, which may contribute to pituitary adenoma development. PMID- 16258075 TI - T-box transcription factors and their roles in regulatory hierarchies in the developing heart. AB - T-box transcription factors are important players in the molecular circuitry that generates lineage diversity and form in the developing embryo. At least seven family members are expressed in the developing mammalian heart, and the human T box genes TBX1 and TBX5 are mutated in cardiac congenital anomaly syndromes. Here, we review T-box gene function during mammalian heart development in the light of new insights into heart morphogenesis. We see for the first time how hierarchies of transcriptional activation and repression involving multiple T-box factors play out in three-dimensional space to establish the cardiac progenitors fields, to define their subservient lineages, and to generate heart form and function. PMID- 16258076 TI - Targeting of amacrine cell neurites to appropriate synaptic laminae in the developing zebrafish retina. AB - Cellular mechanisms underlying the precision by which neurons target their synaptic partners have largely been determined based on the study of projection neurons. By contrast, little is known about how interneurons establish their local connections in vivo. Here, we investigated how developing amacrine interneurons selectively innervate the appropriate region of the synaptic neuropil in the inner retina, the inner plexiform layer (IPL). Increases (ON) and decreases (OFF) in light intensity are processed by circuits that are structurally confined to separate ON and OFF synaptic sublaminae within the IPL. Using transgenic zebrafish in which the majority of amacrine cells express fluorescent protein, we determined that the earliest amacrine-derived neuritic plexus formed between two cell populations whose somata, at maturity, resided on opposite sides of this plexus. When we followed the behavior of individual amacrine cells over time, we discovered that they exhibited distinct patterns of structural dynamics at different stages of development. During cellular migration, amacrine cells exhibited an exuberant outgrowth of neurites that was undirected. Upon reaching the forming IPL, neurites extending towards the ganglion cell layer were relatively more stable. Importantly, when an arbor first formed, it preferentially ramified in either the inner or outer IPL corresponding to the future ON and OFF sublaminae, and maintained this stratification pattern. The specificity by which ON and OFF amacrine interneurons innervate their respective sublaminae in the IPL contrasts with that observed for projection neurons in the retina and elsewhere in the central nervous system. PMID- 16258077 TI - Distinct role of bilobalide and ginkgolide A in the modulation of rat CYP2B1 and CYP3A23 gene expression by Ginkgo biloba extract in cultured hepatocytes. AB - In the present study, primary cultures of rat hepatocytes were treated for 48 h with one of several extracts of Ginkgo biloba (10, 100, or 1000 microg/ml). Maximal increase in CYP2B1 and CYP3A23 mRNA levels was obtained at 100 microg/ml. This concentration of G. biloba extract also increased CYP3A2 and CYP3A18 mRNA expression in addition to CYP2B-mediated 7-benzyloxyresorufin O-dealkylation (BROD) and CYP3A-mediated testosterone 6beta-hydroxylation. In other experiments, cultured hepatocytes were treated for 48 h with bilobalide, ginkgolide A, ginkgolide B, ginkgolide C, ginkgolide J, kaempferol, quercetin, isorhamnetin, or a flavonol diglycoside at a concentration that represented the level present in a 100 microg/ml concentration of an extract. Only bilobalide (2.8 microg/ml) increased CYP2B1 mRNA expression, and the -fold increase (7.9 +/- 0.5; mean +/- S.E.M.) was similar to that (8.3 +/- 1.7) by the extract. By comparison, only ginkgolide A (1.1 microg/ml) increased CYP3A23 mRNA expression, but the extent (2.6 +/- 0.5-fold) was less than the 5.3 +/- 1.7-fold increase by the extract. A greater concentration (5 microg/ml) of ginkgolide A was required to elevate CYP3A2 and CYP3A18 mRNA expression. Over the range of 1 to 5 microg/ml, bilobalide increased CYP2B1 mRNA and BROD, but not CYP3A23 mRNA or testosterone 6beta-hydroxylation, whereas ginkgolide A increased CYP3A23 mRNA and testosterone 6beta-hydroxylation, but not CYP2B1 mRNA or BROD. Overall, our novel results indicate a distinct role of bilobalide and ginkgolide A in the modulation of CYP2B1 and CYP3A23 gene expression and enzyme activities by G. biloba extract in primary cultures of rat hepatocytes. PMID- 16258078 TI - In vivo metabolism of [14C]ruboxistaurin in dogs, mice, and rats following oral administration and the structure determination of its metabolites by liquid chromatography/mass spectrometry and NMR spectroscopy. AB - Ruboxistaurin (LY333531), a potent and isoform-selective protein kinase C beta inhibitor, is currently undergoing clinical trials as a therapeutic agent for the treatment of diabetic microvascular complications. The present study describes the disposition and metabolism of [14C]ruboxistaurin following administration of an oral dose to dogs, mice, and rats. The study revealed that ruboxistaurin was highly metabolized in all species. Furthermore, the results from the bile duct cannulated study revealed that ruboxistaurin was well absorbed in rats. The primary route of excretion of ruboxistaurin and its metabolites was through feces in all species. The major metabolite detected consistently in all matrices for all species was the N-desmethyl metabolite 1, with the exception of rat bile, in which hydroxy N-desmethyl metabolite 5 was detected as the major metabolite. Other significant metabolites detected in dog plasma were 2, 3, 5, and 6 and in mouse plasma 2, 5, and 19. The structures of the metabolites were proposed by tandem mass spectrometry with the exception of 1, 2, 3, 5, and 6, which were additionally confirmed either by direct comparison with authentic standards or by nuclear magnetic resonance spectroscopy. To assist identification by nuclear magnetic resonance spectroscopy, metabolites 3 and 5 were produced via biotransformation using recombinant human CYP2D6 and, likewise, metabolite 6 and compound 4 (regioisomer of 3 which did not correlate to metabolites found in vivo) were produced using a microbe, Mortierella zonata. The unambiguous identification of metabolites enabled the proposal of clear metabolic pathways of ruboxistaurin in dogs, mice, and rats. PMID- 16258079 TI - Glucuronidation of the aspirin metabolite salicylic acid by expressed UDP glucuronosyltransferases and human liver microsomes. AB - Acetylsalicylic acid (aspirin) is a common nonsteroidal anti-inflammatory drug used for treatment of pain and arthritis. In the body, acetylsalicylic acid is rapidly deacetylated to form salicylic acid. Both compounds have been proposed as anti-inflammatory agents. Major metabolites of salicylic acid are its acyl and phenolic glucuronide conjugates. Formation of these conjugates, catalyzed by UDP glucuronosyltransferases (UGTs), decreases the amount of pharmacologically active salicylic acid present. We aimed to identify the UGTs catalyzing the glucuronidation of salicylic acid using both heterologously expressed enzymes and pooled human liver microsomes (HLMs) and to develop a liquid chromatography tandem mass spectrometry method to quantify glucuronidation activity of UGTs 1A1, 1A3, 1A4, 1A6, 1A7, 1A8, 1A9, 1A10, 2B4, 2B7, 2B15, and 2B17 Supersomes. All UGTs tested, except 1A4, 2B15, and 2B17, catalyzed salicylic acid phenolic and acyl glucuronidation. Ratios of salicylic acid phenolic to acyl glucuronide formation varied more than 12-fold from 0.5 for UGT1A6 to 6.1 for UGT1A1. These results suggest that all UGTs except 1A4, 2B15, and 2B17 might be involved in the glucuronidation of salicylic acid in vivo. From comparisons of apparent Km values determined in pooled HLMs and in expressed UGTs, UGT2B7 was suggested as a likely catalyst of salicylic acid acyl glucuronidation, whereas multiple UGTs were suggested as catalysts of phenolic glucuronidation. The results of this UGT screening may help target future evaluation of the effects of UGT polymorphisms on response to aspirin in clinical and population-based studies. PMID- 16258080 TI - Paclitaxel repackaged in an albumin-stabilized nanoparticle: handy or just a dandy? PMID- 16258081 TI - Trastuzumab cardiac side effects: only time will tell. PMID- 16258082 TI - Phase I and pharmacokinetics trial of ABI-007, a novel nanoparticle formulation of paclitaxel in patients with advanced nonhematologic malignancies. AB - PURPOSE: ABI-007 is a novel solvent-free, albumin-bound, 130-nm particle formulation of paclitaxel designed to avoid solvent-related toxicities and to deliver paclitaxel to tumors via molecular pathways involving an endothelial cell surface albumin receptor (gp60) and an albumin-binding protein expressed by tumor cells and secreted into the tumor interstitium (secreted protein acid rich in cysteine). This study determined the maximum-tolerated dose (MTD) of ABI-007 monotherapy administered weekly (three weekly doses, repeated every 4 weeks) and assessed the pharmacokinetics of paclitaxel administered as ABI-007. PATIENTS AND METHODS: Patients with advanced nonhematologic malignancies received ABI-007 without premedication at dose levels from 80 to 200 mg/m(2) as a 30-minute intravenous infusion once a week for 3 weeks, followed by 1 week of rest (one cycle). RESULTS: Thirty-nine patients were treated with an average of five cycles of ABI-007; 33% of patients received > or = six cycles of treatment. MTDs for heavily and lightly pretreated patients were 100 and 150 mg/m(2), respectively; and the dose-limiting toxicities were grade 4 neutropenia and grade 3 peripheral neuropathy, respectively. Maximum paclitaxel concentration and area under the curve increased linearly with dose. Dose-dependent changes in plasma clearance did not occur. Partial responses were observed in five patients with breast, lung, and ovarian cancers, all of whom had previously been treated with paclitaxel containing polyoxyethylated castor oil in the formulation. CONCLUSION: This study demonstrated that weekly ABI-007 can be administered at doses exceeding those typically used for paclitaxel containing polyoxyethylated castor oil. Pharmacokinetics were linear over the dose range studied. Antitumor responses occurred in patients previously treated with paclitaxel containing polyoxyethylated castor oil. PMID- 16258083 TI - Assessment of cardiac dysfunction in a randomized trial comparing doxorubicin and cyclophosphamide followed by paclitaxel, with or without trastuzumab as adjuvant therapy in node-positive, human epidermal growth factor receptor 2-overexpressing breast cancer: NSABP B-31. AB - PURPOSE: Trastuzumab is effective in treating human epidermal growth factor receptor 2 (HER2) -positive breast cancer, but it increases frequency of cardiac dysfunction (CD) when used with or after anthracyclines. PATIENTS AND METHODS: National Surgical Adjuvant Breast and Bowel Project trial B-31 compared doxorubicin and cyclophosphamide (AC) followed by paclitaxel with AC followed by paclitaxel plus 52 weeks of trastuzumab beginning concurrently with paclitaxel in patients with node-positive, HER2-positive breast cancer. Initiation of trastuzumab required normal post-AC left ventricular ejection fraction (LVEF) on multiple-gated acquisition scan. If symptoms suggestive of congestive heart failure (CHF) developed, source documents were blindly reviewed by an independent panel of cardiologists to determine whether criteria were met for a cardiac event (CE), which was defined as New York Heart Association class III or IV CHF or possible/probable cardiac death. Frequencies of CEs were compared between arms. RESULTS: Among patients with normal post-AC LVEF who began post-AC treatment, five of 814 control patients subsequently had confirmed CEs (four CHFs and one cardiac death) compared with 31 of 850 trastuzumab-treated patients (31 CHFs and no cardiac deaths). The difference in cumulative incidence at 3 years was 3.3% (4.1% for trastuzumab-treated patients minus 0.8% for control patients; 95% CI, 1.7% to 4.9%). Twenty-seven of the 31 patients in the trastuzumab arm have been followed for > or = 6 months after diagnosis of a CE; 26 were asymptomatic at last assessment, and 18 remained on cardiac medication. CHFs were more frequent in older patients and patients with marginal post-AC LVEF. Fourteen percent of patients discontinued trastuzumab because of asymptomatic decreases in LVEF; 4% discontinued trastuzumab because of symptomatic cardiotoxicity. CONCLUSION: Administering trastuzumab with paclitaxel after AC increases incidence of CHF and lesser CD. Potential cardiotoxicity should be carefully considered when discussing benefits and risks of this therapy. PMID- 16258084 TI - Reversibility of trastuzumab-related cardiotoxicity: new insights based on clinical course and response to medical treatment. AB - PURPOSE: Trastuzumab is an important biologic agent with significant activity in breast cancers that overexpress the HER2/neu marker. However, trastuzumab is associated with cardiotoxicity that has not yet been fully explored. We present our experience with patients who developed trastuzumab-related cardiotoxicity. PATIENTS AND METHODS: Over a 4-year period, 38 patients with HER2/neu-positive breast cancer were referred for suspected trastuzumab-related cardiotoxicity. All patients had previously received anthracycline-based chemotherapy. Results After doxorubicin but before trastuzumab, the mean (+/- standard deviation) left ventricular ejection fraction (LVEF) was 0.61 +/- 0.13, and the LVEF decreased to 0.43 +/- 0.16 after trastuzumab (P < .0001). After withdrawal of trastuzumab, the LVEF increased to 0.56 +/- 0.11. Mean time to recovery of LVEF was 1.5 months and was temporally associated with medical treatment in 32 (84%) of the 38 patients but occurred without treatment in six patients (16%). Increases in LVEF were noted in 37 of the 38 patients. Twenty-five of these patients were re-treated with trastuzumab; three patients had recurrent left ventricular dysfunction, but 22 patients (88%) did not. All re-treatment patients continued on their therapeutic regimen for heart failure when rechallenged with trastuzumab. Nine patients underwent endomyocardial biopsy. Ultrastructural changes were not seen. CONCLUSION: Patients who develop cardiotoxicity while receiving trastuzumab therapy generally improve on removal of the agent. The mechanism of trastuzumab related cardiac dysfunction is different from that of anthracycline cardiotoxicity, in part, demonstrated by the absence of anthracycline-like ultrastructural changes. Reintroducing trastuzumab may be appropriate for some individuals who previously have experienced trastuzumab-related cardiac dysfunction. PMID- 16258085 TI - Squamous cell carcinoma of the breast. AB - PURPOSE: Squamous cell carcinoma (SCC) of the breast is rare and generally aggressive. In this study, we analyzed 33 patients treated at The University of Texas M.D. Anderson Cancer Center (Houston, TX) and a series of 137 patients identified through the Surveillance, Epidemiology, and End Results (SEER) database to make therapy recommendations. METHODS: Records of The University of Texas M.D. Anderson Cancer Center were searched for patients diagnosed with breast SCC from 1985 to 2001. All biopsy material was reviewed by a dedicated breast pathologist who performed immunohistochemistry for hormone receptors and the epidermal growth factor receptor (EGFR). We searched the SEER database for patients with breast SCC diagnosed between 1988 and 2001. RESULTS: We identified 33 patients with breast SCC, of whom two patients had metastatic disease at diagnosis. The median relapse-free survival (RFS) of 31 patients with localized disease was 20 months (range, 1 to 108 months), with a 26% RFS rate at 5 years. The median overall survival in these patients was 37 months (range, 12 to 108 months), with 40% surviving at 5 years. Median survival from the time recurrent disease was recognized was 14 months (range, 2 to 86 months). Tumors were usually hormone receptor-and HER2/neu-negative, though EGFR was frequently overexpressed. Information from the SEER database was consistent with most of our findings. CONCLUSION: SCC of the breast is aggressive and often treatment-refractory. The role of platinum salts, EGFR inhibitors, and other novel agents needs to be explored. PMID- 16258086 TI - Breast cancer trends among black and white women in the United States. AB - PURPOSE: Overall US breast cancer mortality rates are higher among black women than white women, and the disparity is widening. To investigate this disparity, we examined incidence data and changes in mortality trends according to age, year of death (calendar period), and date of birth (birth cohort). Calendar period mortality trends reflect the effects of new medical interventions, whereas birth cohort mortality trends reflect alterations in risk factors. PATIENTS AND METHODS: Incidence data were obtained from the Connecticut and National Cancer Institute Surveillance, Epidemiology, and End Results registries and mortality data were obtained from the National Center for Health Statistics. Changes in age, period, and cohort mortality trends were analyzed with Poisson regression. RESULTS: For both races, breast cancer incidence rates for localized and regional disease diverged in the late 1970s. Almost concurrently, overall mortality rates diverged among blacks and whites. For both races, mortality increases with age, but blacks have higher mortality at age younger than 57. The calendar period curves revealed declining mortality for whites over the entire study period. For blacks, calendar period mortality declined until the late 1970s, and then sharply increased. After 1994, calendar period mortality declined for both. For women born between 1872 and 1950, trends in mortality were similar for blacks and whites. For women born after 1950, mortality decreased more rapidly for blacks. CONCLUSION: The widening racial disparity in breast cancer mortality seems attributable to calendar period rather than birth cohort effects. Thus, differences in response or access to newer medical interventions may largely account for these trends. PMID- 16258087 TI - Tamoxifen versus control after adjuvant, risk-adapted chemotherapy in postmenopausal, receptor-negative patients with breast cancer: a randomized trial (GABG-IV D-93)--the German Adjuvant Breast Cancer Group. AB - PURPOSE: To investigate the effect of adjuvant sequential tamoxifen after chemotherapy in postmenopausal patients with hormone receptor-negative breast cancer. METHODS: Patients were randomly assigned to oral tamoxifen (30 mg daily for 5 years; n = 421) or no additional treatment (n = 408) after risk-adapted polychemotherapy consisting of three 28-day cycles of CMF (cyclophosphamide, 500 mg/m(2), methotrexate, 40 mg/m(2), and fluorouracil, 600 mg/m(2)) in patients with negative or one to three positive lymph nodes and four 21-day cycles of epirubicin 90 mg/m(2), cyclophosphamide 600 mg/m(2) followed by three cycles of CMF in patients with four to nine positive lymph nodes. RESULTS: Thirty-six percent of the patients included were older than 60 years, 63% were node negative, 13% had four to nine positive nodes, 55% had tumor grade 3, and 41% received breast-preserving surgery. At 5.3 years' median follow-up, the first event of failure (recurrence, secondary tumor, or death) had occurred in 123 patients in the tamoxifen group and 107 patients of the control group. Event-free survival rates after 5 years were 70.3% (95% CI, 65.5% to 75.0%) and 72.8% (95% CI, 68.2% to 77.5%) for the tamoxifen and control groups, respectively. The estimated hazard ratio of tamoxifen versus control was 1.13 (95% CI, 0.87 to 1.48; P = .34), which gives no indication of an additional benefit of tamoxifen in these patients. CONCLUSION: This study contributes substantially to finalization of the presently emerging evidence that tamoxifen does not benefit women with receptor-negative breast cancer after chemotherapy. PMID- 16258088 TI - Randomized trial of prevention of catheter-related bloodstream infection by continuous infusion of low-dose unfractionated heparin in patients with hematologic and oncologic disease. AB - PURPOSE: Infection is a serious complication of central venous catheters in immunocompromised patients. Catheter-related infection may be caused by fibrin deposition associated with catheters. Interventions designed to decrease fibrin deposition have the potential to reduce catheter-related infections. The purpose of this study was to evaluate the role of low-dose unfractionated heparin in preventing catheter-related bloodstream infection in patients with hemato oncological disease. PATIENTS AND METHODS: This study was a randomized, controlled trial in which patients with nontunneled catheters were randomly assigned to receive either intravenous unfractionated heparin (continuous infusion of 100 U/kg per day) or 50 mL/day of normal saline solution as a continuous infusion (control group). Heparin was continued until the day of discharge. Catheter-related bloodstream infection was defined according to Infectious Disease Society of America guidelines. RESULTS: Two hundred and eight patients were randomly assigned. Four patients were excluded after assignment. Ultimately, 204 patients were analyzed. Catheter-related bloodstream infection occurred in 6.8% (7 of 102 catheters) of those in the heparin group (2.5 events per 1,000 days) and in 16.6% (17 of 102 catheters) of those in the control group (6.4 events per 1,000 days) (P = .03). No other risk factors were found for the development of catheter-related bloodstream infection. Four and five patients experienced severe bleeding in the heparin and control groups, respectively (P = .2). We did not observe heparin-induced thrombocytopenia. CONCLUSION: The use of continuous infusion of low-dose unfractionated heparin (100 U/kg per day) can be a practical and economical approach to the prevention of catheter-related bloodstream infection in patients with hemato-oncological disease. PMID- 16258089 TI - Gonadotropin-releasing hormone agonists and fracture risk: a claims-based cohort study of men with nonmetastatic prostate cancer. AB - PURPOSE: Gonadotropin-releasing hormone (GnRH) agonists decrease bone mineral density, a surrogate for fracture risk, in men with prostate cancer. We conducted a claims-based cohort study to characterize the relationship between GnRH agonists and risk for clinical fractures in men with nonmetastatic prostate cancer. PATIENTS AND METHODS: Using medical claims data from a 5% national random sample of Medicare beneficiaries, we identified a study group of men with nonmetastatic prostate cancer who initiated GnRH agonist treatment from 1992 to 1994 (n = 3,887). A comparison group of men with nonmetastatic prostate cancer who did not receive GnRH agonist treatment during the study period (n = 7,774) was matched for age, race, geographic location, and comorbidity. Clinical fractures were identified using inpatient, outpatient, and physician claims during 7 years of follow-up. RESULTS: In men with nonmetastatic prostate cancer, GnRH agonists significantly increased fracture risk. The rate of any clinical fracture was 7.88 per 100 person-years at risk in men receiving a GnRH agonist compared with 6.51 per 100 person-years in matched controls (relative risk, 1.21; 95% CI, 1.14 to 1.29; P < .001). Rates of vertebral fractures (relative risk, 1.45; 95% CI, 1.19 to 1.75; P < .001) and hip/femur fractures (relative risk, 1.30; 95% CI, 1.10 to 1.53; P = .002) were also significantly higher in men who received a GnRH agonist. GnRH agonist treatment independently predicted fracture risk in multivariate analyses. Longer duration of treatment conferred greater fracture risk. CONCLUSION: GnRH agonists significantly increase risk for any clinical fracture, hip fractures, and vertebral fractures in men with prostate cancer. PMID- 16258090 TI - Therapy tolerance in selected patients with androgen-independent prostate cancer following strontium-89 combined with chemotherapy. AB - PURPOSE: Clinicians may have reservations about using strontium-89 for the treatment of bone metastases because of concerns that it may limit future use of chemotherapy. We assessed the rate of bone marrow failure in patients with prostate cancer who had received a dose of strontium-89. PATIENTS AND METHODS: This subgroup analysis involved 34 patients with androgen-independent prostate cancer who had been given a dose of strontium-89 and six weekly doses of doxorubicin after response to induction chemotherapy. We assessed subsequent hematotoxicity in terms of bone marrow failure and the ability to tolerate additional treatments during a median of 25 months (range, 7 to 76 months) after the strontium-89 was administered. RESULTS: No patients developed bone marrow failure within 6 months of receiving strontium-89. Five (15%) of 34 patients developed bone marrow failure at a median 23 months (range, 6 to 53 months) after the strontium-89 treatment. Bone marrow biopsy performed in two of these five patients showed complete replacement of the marrow by tumor. Thirty-one patients (91%) received subsequent cytotoxic treatments at a median 11 months (range, 1 to 33 months) after the strontium-89 treatment. CONCLUSION: This analysis demonstrated that a single dose of strontium-89 combined with chemotherapy did not affect the delivery of subsequent courses of chemotherapy in a select group of patients. However, a majority of these therapies were given off protocol and were administered at a dose schedule that might be considered inappropriate or inadequate. The clinical role and safety profile of radiopharmaceuticals combined with chemotherapy in prostate cancer therapy deserve further exploration. PMID- 16258091 TI - Prospective study using the risk of ovarian cancer algorithm to screen for ovarian cancer. AB - PURPOSE: To evaluate prevalence screening in the first prospective trial of a new ovarian cancer screening (OCS) strategy (risk of ovarian cancer or ROC algorithm) on the basis of age and CA125 profile. PATIENTS AND METHODS: Postmenopausal women, > or = 50 years were randomly assigned to a control group or screen group. Screening involved serum CA125, interpreted using the ROC algorithm. Participants with normal results returned to annual screening; those with intermediate results had repeat CA125 testing; and those with elevated values underwent transvaginal ultrasound (TVS). Women with abnormal or persistently equivocal TVS were referred for a gynecologic opinion. RESULTS: Thirteen thousand five hundred eighty-two women were recruited. Of 6,682 women randomly assigned to screening, 6,532 women underwent the first screen. After the initial CA125, 5,213 women were classified as normal risk, 91 women elevated, and 1,228 women intermediate. On repeat CA125 testing of the latter, a further 53 women were classified as elevated risk. All 144 women with elevated risk had TVS. Sixteen women underwent surgery. Eleven women had benign pathology; one woman had ovarian recurrence of breast cancer; one woman had borderline; and three women had primary invasive epithelial ovarian cancer (EOC). The specificity and positive predictive value (PPV) for primary invasive EOC were 99.8% (95% CI, 99.7 to 99.9) and 19% (95% CI, 4.1 to 45.6), respectively. CONCLUSION: An OCS strategy using the ROC algorithm is feasible and can achieve high specificity and PPV in postmenopausal women. It is being used in the United Kingdom Collaborative Trial of Ovarian Cancer Screening and in the United States in both the Cancer Genetics Network and the Gynecology Oncology Group trials of high-risk women. PMID- 16258092 TI - Response of retinoblastoma with vitreous tumor seeding to adenovirus-mediated delivery of thymidine kinase followed by ganciclovir. AB - PURPOSE: To evaluate the feasibility and safety of adenovirus-mediated gene therapy as a treatment for tumor seeds in the vitreous of children with retinoblastoma. PATIENTS AND METHODS: An Institutional Biosafety Committee-, Institutional Review Board-, Recombinant DNA Advisory Committee-, and US Food and Drug Administration-approved phase I study used intrapatient dose escalation of adenoviral vector containing a herpes simplex thymidine kinase gene (AdV-TK) followed by systemic administration of ganciclovir to treat bilateral retinoblastoma with vitreous tumor seeding refractory to standard therapies. Vitreous tumor seeds were treated by intravitreous injection of AdV-TK adjacent to disease sites. Each injection was followed by ganciclovir delivered intravenously every 12 hours for 7 days. RESULTS: Eight patients with vitreous tumor seeds were enrolled. One patient who was treated with 10(8) viral particles (vp) had resolution of the tumor seeds around the injection site. The seven patients who were treated with doses > or = 10(10) vp had resolution of their vitreous tumor seeds documented by fundoscopy. Toxicity included mild inflammation at 10(10) vp and moderate inflammation, corneal edema, and increased intraocular pressure at 10(11) vp. One patient was free of active vitreous tumor seeds 38 months after therapy. There has been no evidence of extraocular spread of tumor along the needle tract in any patient. CONCLUSION: AdV-TK followed by ganciclovir can be administered safely to children with retinoblastoma. Suicide gene therapy may contribute to the treatment of children with retinoblastoma tumor seeds in the vitreous, a resistant complication of retinoblastoma. PMID- 16258093 TI - Risk of adverse events after completion of therapy for childhood acute lymphoblastic leukemia. AB - PURPOSE: We studied the frequency, causes, and predictors of adverse events in children with acute lymphoblastic leukemia (ALL) who had completed treatment on contemporary clinical protocols between 1984 and 1999. Our goal was to use the information to further refine therapy and advance cure rates. METHODS: Cumulative incidence functions of any post-treatment failure or any post-treatment relapse were estimated by the method of Kalbfleisch and Prentice and compared with Gray's test. The Cox proportional hazards model was used to identify independent prognostic factors. RESULTS: Of the 827 patients who completed all treatment while in initial complete remission, 134 patients subsequently had major adverse events, including 90 leukemic relapses, 40 second malignancies, and four deaths in remission. The cumulative incidence of any adverse event was 14.0% +/- 1.2% (SE) at 5 years and 16.9% +/- 1.4% at 10 years. The risk of any leukemic relapse was 10.0% +/- 1.1% at 5 years and 11.4% +/- 1.2% at 10 years. Male sex was the only independent predictor of relapse (hazard ratio, 1.74; 95% CI, 1.11 to 2.74; P = .02). CONCLUSION: Further treatment refinements for children with ALL should aim not only to decrease the leukemic relapse rate, but also to reduce the risk of development of second malignancies. PMID- 16258094 TI - Long-term outcome in children with relapsed ALL by risk-stratified salvage therapy: results of trial acute lymphoblastic leukemia-relapse study of the Berlin-Frankfurt-Munster Group 87. AB - PURPOSE: Approximately 20% of children with acute lymphoblastic leukemia (ALL) suffer a relapse, and their prognosis is unfavorable. Between 1987 and 1990, the multicenter trial Acute Lymphoblastic Leukemia-Relapse Study of the Berlin Frankfurt-Munster Group (ALL-REZ BFM) 87 was conducted to establish a uniform treatment for these children in Germany and Austria. PATIENTS AND METHODS: Of 207 registered patients, 183 patients were stratified into three groups according to the protocol: A, early bone marrow (BM) relapse (n = 56); B, late BM relapse (n = 101); C, isolated extramedullary relapse (n = 26). Treatment consisted of risk adapted alternating short-course multiagent systemic and intrathecal chemotherapy, cranial irradiation, if indicated, and conventional maintenance therapy. Additionally, 24 patients with an exceptionally poor prognosis (early BM or any relapse of T-cell ALL) were treated with individual regimens. In 35 patients, stem-cell transplantation was performed. RESULTS: The probability of event-free survival (EFS) and overall survival of all registered patients at 15 years was 0.30 +/- 0.03 and 0.37 +/- 0.03, respectively, with significant differences between the strategic groups (A, 0.18 +/- 0.05 and 0.20 +/- 0.05; B, 0.44 +/- 0.05 and 0.52 +/- 0.05; C, 0.35 +/- 0.09 and 0.42 +/- 0.10). Despite risk-adapted treatment, an early time point of relapse and T-lineage immunophenotype were significant predictors of inferior EFS in uni- and multivariate analyses. CONCLUSION: With the ALL-REZ BFM 87 protocol, more than one-third of patients may be regarded as cured from recurrent ALL with second complete remissions lasting more than 10 years. Immunophenotype and time point of relapse are important prognostic factors that allow us to adapt more precisely treatment intensity to individual prognosis in future trials. PMID- 16258095 TI - beta-Catenin status predicts a favorable outcome in childhood medulloblastoma: the United Kingdom Children's Cancer Study Group Brain Tumour Committee. AB - PURPOSE: Identifying pathobiological correlates of clinical behavior or therapeutic response currently represents a key challenge for medulloblastoma research. Nuclear accumulation of the beta-catenin protein is associated with activation of the Wnt/Wg signaling pathway, and mutations affecting components of this pathway have been reported in approximately 15% of sporadic medulloblastomas. We tested the hypothesis that nuclear immunoreactivity for beta catenin is a prognostic marker in medulloblastoma, and assessed the relationship between nuclear beta-catenin immunoreactivity and mutations of CTNNB1 and APC. PATIENTS AND METHODS: Medulloblastomas from children entered onto the International Society for Pediatric Oncology (SIOP)/United Kingdom Children's Cancer Study Group (UKCCSG) PNET3 trial (n = 109) were examined for beta-catenin immunoreactivity, and where tissue was available, evidence of CTNNB1 and APC mutations. The results were correlated with clinicopathologic variables, principally outcome. RESULTS: Children with medulloblastomas that showed a nucleopositive beta-catenin immunophenotype (27 of 109; 25%) had significantly better overall (OS) and event-free (EFS) survivals than children with tumors that showed either membranous/cytoplasmic beta-catenin immunoreactivity or no immunoreactivity (P = .0015 and P = .0026, respectively). For beta-catenin nucleopositive and nucleonegative medulloblastomas, 5-year OS was 92.3% (95% CI, 82% to 100%) versus 65.3% (95% CI, 54.8 to 75.7%), and 5-year EFS was 88.9% (95% CI, 77% to 100%) versus 59.5% (95% CI, 48.8 to 70.2%), respectively. Mutations in CTNNB1 were found exclusively among medulloblastomas that demonstrated nuclear beta-catenin immunoreactivity, but no evidence of APC mutation was found in these cases. All children with beta-catenin nucleopositive large cell/anaplastic medulloblastomas and beta-catenin nucleopositive medulloblastomas presenting with metastatic disease are alive at least 5 years postdiagnosis. CONCLUSION: Nuclear accumulation of beta-catenin appears to be a marker of favorable outcome in medulloblastoma, and should be investigated further in large group-wide trials. PMID- 16258096 TI - Length of stay and mortality associated with febrile neutropenia among children with cancer. AB - PURPOSE: The aim of this study was to evaluate risk factors for longer length of stay (los) and mortality among hospitalized children with cancer who have febrile neutropenia. METHODS: This study involved analysis of longitudinal data from the University HealthSystem Consortium database from 1995 to 2002. All patients who were 21 years or younger, with diagnostic codes for both neoplastic disease and febrile neutropenia at discharge, were included. RESULTS: A total of 12,446 patients were identified for the study. The los was 5 days or less for 6,799 patients, and greater than 5 days for 5,647 patients. The mortality rate was 3%. On bivariate analysis, race, age, cancer type, and associated complications (bacteremia/sepsis, hypotension, pneumonia, and fungal infections) were significantly associated with longer length of stay and death. On multivariate analysis, age group, race, cancer type (acute myeloid leukemia, multiple cancers v acute lymphoblastic leukemia), and the complication variables were significantly associated with increased risk of longer los and death. Certain types of cancer (Hodgkin's disease, osteosarcoma/Ewing's sarcoma, rhabdomyosarcoma, compared with acute lymphoblastic leukemia) and year of discharge after 1995 were significantly associated with a reduced risk of longer length of stay and/or mortality. CONCLUSION: Race, age group, year of discharge, associated complications, and cancer type were significantly associated with risk of longer los and mortality. These factors may potentially help in identifying high-risk patients who might benefit from targeted antibiotic therapy or prophylactic hematopoietic growth factor support. PMID- 16258097 TI - Recruiting African American women to participate in hereditary breast cancer research. AB - PURPOSE: This study evaluated the process of recruiting African American women to participate in genetic counseling research for BRCA1 and BRCA2 (BRCA1/2) mutations with respect to referral, study enrollment, and participation in genetic counseling. PATIENTS AND METHODS: African American women (n = 783) were referred for study enrollment. RESULTS: Of 783 referrals, 164 (21%) women were eligible for enrollment. Eligible women were most likely to be referred from oncology clinics (44%) and were least likely to be referred from general medical practices (11%; chi(2) = 96.80; P = .0001). Overall, 62% of eligible women enrolled onto the study and 50% of enrollees completed genetic counseling. Women with a stronger family history of cancer (odds ratio [OR] = 3.18; 95% CI, 1.36 to 7.44; P = .01) and those referred from oncology clinics and community oncology resources (OR = 2.97; 95% CI, 1.34 to 6.58; P = .01) were most likely to enroll onto the study. Referral from oncology clinics was associated significantly with participation in genetic counseling (OR = 5.46; 95% CI, 1.44 to 20.60; P = .01). CONCLUSION: Despite receiving a large number of referrals, only a small subset of women were eligible for enrollment. Oncology settings were the most effective at identifying eligible African American women and general medical practices were the least effective. Factors associated with enrollment included having a stronger family history of cancer and being referred from oncology clinics and community oncology resources. Referral from oncology clinics was the only factor associated significantly with participation in genetic counseling. Education about hereditary breast cancer may be needed among primary care providers to enhance appropriate referral of African American women to genetic counseling for BRCA1/2 mutations. PMID- 16258098 TI - Prevention of chemotherapy-induced febrile neutropenia by prophylactic antibiotics plus or minus granulocyte colony-stimulating factor in small-cell lung cancer: a Dutch Randomized Phase III Study. AB - PURPOSE: Febrile neutropenia (FN) is a major complication of chemotherapy. Antibiotics as well as granulocyte colony-stimulating factor (G-CSF) are effective in preventing FN. This multicenter randomized phase III trial determines whether the addition of G-CSF to antibiotic prophylaxis can further reduce the incidence of FN in patients with small-cell lung cancer (SCLC) at the risk of FN. PATIENTS AND METHODS: Patients (N = 175) were stratified for stage of disease, performance status, age, and prior chemotherapy treatment, and were randomly assigned for treatment with cyclophosphamide, doxorubicin, and etoposide (CDE), followed by prophylactic antibiotics alone (ciprofloxacin and roxithromycin) or by antibiotics in combination with G-CSF on days 4 to 13. RESULTS: In cycle 1, 20 patients (24%) in the antibiotics group developed FN compared with nine patients (10%) in the antibiotics plus G-CSF group (P = .01). In cycles 2 to 5, the incidences of FN were practically the same in both groups (17% v 11%). Only the treatment parameters (odds ratio, 0.33; 95% CI, 0.14 to 0.78) and age (1.067 per year; 95% CI, 1.013 to 1.0124) were related to the probability of FN in cycle 1. CONCLUSION: Primary G-CSF prophylaxis added to primary antibiotic prophylaxis is effective in reducing FN and infections in SCLC patients at the risk of FN with the first cycle of CDE chemotherapy. For patients with similar risk of FN, the combined use of prophylactic antibiotics plus G-CSF can be considered, specifically in the first cycle of chemotherapy. PMID- 16258099 TI - Prognostic significance of BACH2 expression in diffuse large B-cell lymphoma: a study of the Osaka Lymphoma Study Group. AB - PURPOSE: BACH2, a B-cell-specific transcription repressor, is abundantly expressed in lymphocytes of B-cell lineage as well as B-cell lymphoma cell lines. BACH2 possesses an inhibitory effect on proliferation of Raji cell lines derived from Burkitt's lymphoma. In this study, the prognostic significance of BACH2 expression was examined in diffuse large B-cell lymphoma (DLBCL). PATIENTS AND METHODS: BACH2 expression was immunohistochemically examined on the paraffin embedded sections obtained by biopsy from 108 patients (62 males and 46 females; age range, 23 to 85 years) with DLBCL. Staining intensity in the cytoplasm of the tumor cells was categorized as equal to or stronger (level 1) or weaker (level 2) than that in the endothelial cells in the same specimens. RESULTS: Level 1 and 2 expression of BACH2 was found in 32.4% and 67.6% of patients, respectively. Patients with level 1 expression showed significantly better disease-free and overall survival rate than those with level 2 expression (both P < .05). Multivariate analysis revealed BACH2 expression level together with performance status, elevated serum level of lactate dehydrogenase, and treatment response to be independent factors for prognosis of the patients. CONCLUSION: BACH2 expression level is a useful marker to predict disease-free and overall survival of patients with DLBCL. PMID- 16258100 TI - Fatigue, menopausal symptoms, and cognitive function in women after adjuvant chemotherapy for breast cancer: 1- and 2-year follow-up of a prospective controlled study. AB - PURPOSE: We previously evaluated fatigue, menopausal symptoms, and cognitive dysfunction in patients receiving adjuvant therapy for breast cancer and matched healthy women. Here we report assessment of these women 1 and 2 years later. PATIENTS AND METHODS: Patients without relapse and controls were evaluated by the Functional Assessment of Cancer Treatment-General Quality of Life questionnaire, with subscales for fatigue and endocrine symptoms, and by the High Sensitivity Cognitive Screen. RESULTS: There were 104, 91, and 83 patients and 102, 81, and 81 controls assessed at baseline and at 1 and 2 years, respectively. Median Functional Assessment of Cancer Treatment-Fatigue scores (range, 0 to 52) for patients improved from 31 (on chemotherapy) to 43 and 45 at 1 and 2 years, respectively, but were stable in controls (46 to 48). Median Functional Assessment of Cancer Treatment-Endocrine Symptoms scores (range, 0 to 72) for patients improved from 57 (on chemotherapy) to 59 and 61 at 1 and 2 years, respectively, and were stable in controls (64 to 65). Differences between patients and controls remained significant for these scales. The incidence of moderate-severe cognitive dysfunction by the High Sensitivity Cognitive Screen decreased in patients from 16% (on chemotherapy) to 4.4% and 3.8% and in controls from 5% to 3.6% and 0% at 1 and 2 years, respectively. There were minimal differences between estrogen receptor-positive patients who started hormonal therapy (mainly tamoxifen) after chemotherapy and estrogen receptor-negative patients who did not. Differences in quality of life between patients and controls were significant only at baseline. CONCLUSION: Fatigue, menopausal symptoms, and cognitive dysfunction are important adverse effects of chemotherapy that improve in most patients. Hormonal treatment has minimal impact on them. PMID- 16258101 TI - Phase II trial of bevacizumab plus gemcitabine in patients with advanced pancreatic cancer. AB - PURPOSE: Vascular endothelial growth factor (VEGF) plays a key role in the biology and prognosis of pancreatic cancer. Inhibitors of VEGF suppress the growth of pancreatic cancer in preclinical models. The objectives of this phase II study were to assess the response rate and overall survival of pancreatic cancer patients who received gemcitabine with the recombinant humanized anti-VEGF monoclonal antibody bevacizumab. PATIENTS AND METHODS: Patients with previously untreated advanced pancreatic cancer received gemcitabine 1,000 mg/m(2) intravenously over 30 minutes on days 1, 8, and 15 every 28 days. Bevacizumab, 10 mg/kg, was administered after gemcitabine on days 1 and 15. Tumor measurements were assessed every two cycles. Plasma VEGF levels were obtained pretreatment. RESULTS: Fifty-two patients were enrolled at seven centers between November 2001 and March 2004. All patients had metastatic disease, and 83% had liver metastases. Eleven patients (21%) had confirmed partial responses, and 24 (46%) had stable disease. The 6-month survival rate was 77%. Median survival was 8.8 months; median progression-free survival was 5.4 months. Pretreatment plasma VEGF levels did not correlate with outcome. Grade 3 and 4 toxicities included hypertension in 19% of the patients, thrombosis in 13%, visceral perforation in 8%, and bleeding in 2%. CONCLUSION: The combination of bevacizumab plus gemcitabine is active in advanced pancreatic cancer patients. Additional study is warranted. A randomized phase III trial of gemcitabine plus bevacizumab versus gemcitabine plus placebo is ongoing in the Cancer and Leukemia Group B. PMID- 16258102 TI - An intensive surveillance program detected a high incidence of hepatocellular carcinoma among hepatitis B virus carriers with abnormal alpha-fetoprotein levels or abdominal ultrasonography results. AB - PURPOSE: To study the incidence and treatment outcomes of hepatocellular carcinoma (HCC) detected in an intensive surveillance program (ISP) of hepatitis B virus (HBV) carriers. PATIENTS AND METHODS: We screened 1,018 HBV carriers by serum alpha-fetoprotein (AFP) measurement and abdominal ultrasonography (AUS). Patients with an abnormal AFP level or AUS result were enrolled in an ISP that included Lipiodol computed tomography followed by AFP measurement/AUS every 3 months for 2 years and then every 6 months thereafter. The rest were on routine surveillance for 2 years. RESULTS: A total of 9,849 serum AFP measurements and 3,053 AUSs were performed. After a median follow-up of 4.12 years, we diagnosed 24 HCCs among 78 patients with abnormal screening test results at enrollment (group A); 23 HCCs among 93 patients with only abnormal surveillance test results during follow-up (group B); and nine HCCs among 847 patients with 2 years of normal surveillance (group C). Annual incidence of HCC in the ISP was 760.2 (95% CI, 538.4 to 1,073.7) per 100,000. Mean tumor sizes were 3.02, 2.91, and 4.82 cm in groups A, B, and C, respectively (P = .01). Tumor resection rate of the ISP was 36.2%, although another 29.8% of the patients were eligible for locoregional ablative therapy. CONCLUSION: This study illustrated that a high incidence of relatively small HCCs may be detected by using intensive surveillance of high risk HBV carriers. However, the surgical resection rate was low, and we were not able to demonstrate clinical benefit with the early detection. Future surveillance studies should consider incorporation of therapy aimed at long-term control of small-sized tumors. PMID- 16258103 TI - Biologic and prognostic significance of dermal Ki67 expression, mitoses, and tumorigenicity in thin invasive cutaneous melanoma. AB - PURPOSE: Tumor cell proliferation is a central feature of melanoma progression. In this study, we characterized three biomarkers of proliferation (Ki67 expression, dermal mitotic rate [MR], and tumorigenicity) in thin (< or = 1.00 mm) primary cutaneous melanomas and examined their association with prognosis. PATIENTS AND METHODS: We used immunohistochemistry to determine Ki67 expression using the monoclonal antibody MIB-1 in lesions from a prospective cohort that included 396 patients with thin invasive primary melanomas seen between 1972 and 1991. A multivariate Cox proportional hazards model was used to define independent prognostic factors, and recursive partitioning was used to develop a prognostic tree identifying risk groups. RESULTS: Dermal Ki67 expression was lower than epidermal Ki67 expression in radial growth phase (RGP) melanomas (n = 171), and dermal Ki67 expression and MR were higher in tumorigenic vertical growth phase (VGP) melanomas (n = 193) compared with RGP and nontumorigenic VGP melanomas (n = 42). Dermal Ki67 expression, MR greater than 0, growth phase, thickness, ulceration, tumor-infiltrating lymphocytes, and sex were associated with metastasis at 10 years, however, only dermal Ki67 expression, MR greater than 0, and sex were independent prognostic factors. Two high-risk groups were identified: men and women with dermal MR greater than 0 and dermal Ki67 expression > or = 20% in tumor cells and men with MR greater than 0 and Ki67 expression less than 20%, with 10-year metastasis rates of 39% and 20%, respectively. CONCLUSION: Proliferation slows as melanoma cells enter the dermis and then increases with the onset of tumorigenic VGP. Ki67 expression and dermal MR provide independent prognostic information that can potentially be used in risk-based management of patients. PMID- 16258104 TI - Serial monitoring of circulating melanoma cells during neoadjuvant biochemotherapy for stage III melanoma: outcome prediction in a multicenter trial. AB - PURPOSE: Circulating tumor cells (CTCs) in blood may be important in assessing tumor progression and treatment response. We hypothesized that quantitative real time reverse transcriptase polymerase chain reaction using multimarker mRNA assays could detect CTCs and be used as a surrogate predictor of outcome in patients receiving neoadjuvant biochemotherapy (BC) for melanoma. PATIENTS AND METHODS: Blood specimens were collected at four sampling points from 63 patients enrolled on a prospective multicenter phase II trial of BC before and after surgical treatment of American Joint Committee on Cancer stage III melanoma. Each specimen was assessed by quantitative real-time reverse transcriptase polymerase chain reaction for expression of four melanoma-associated markers: melanoma antigen recognized by T cells 1; beta1 --> 4-N-acetylgalactosaminyltransferase; paired box homeotic gene transcription factor 3; and melanoma antigen gene-A3 family, and the changes of CTCs during treatment and prognostic effect of CTCs after overall treatment on recurrence and survival were investigated. RESULTS: At a median postoperative follow-up time of 30.4 months, 44 (70%) patients were clinically disease free. In relapse-free patients, the number of detected markers significantly decreased during preoperative BC (P = .036), during postoperative BC (P = .002), and during overall treatment (P < .0001). Marker detection after overall treatment was associated with significant decreases in relapse-free and overall survival (P < .0001). By multivariate analysis using a Cox proportional hazards model, the number of markers detected after overall treatment was a significant independent prognostic factor for overall survival (risk ratio, 12.6; 95% CI, 3.16 to 50.5; P = .0003). CONCLUSION: Serial monitoring of CTCs in blood may be useful for indicating systemic subclinical disease and predicting outcome of patients receiving neoadjuvant BC for metastatic melanoma. PMID- 16258105 TI - A population-based validation of the American Joint Committee on Cancer melanoma staging system. AB - PURPOSE: A major revision of the American Joint Committee on Cancer (AJCC) stages for melanoma was implemented in 2002 after its validation in multinational cohorts including patients from cancer centers and cooperative groups. This staging system has not been validated in a US population-based cohort. PATIENTS AND METHODS: We used 41,417 patients with primary invasive cutaneous melanoma diagnosed between 1988 and 2001 from the National Cancer Institute's Surveillance, Epidemiology, and End Results (SEER) cancer registry to validate the revised AJCC staging system. Survival rates computed from stage-specific Kaplan-Meier curves (time to melanoma-specific death) were compared with the survival rates from 17,600 patients in the original AJCC validation study. RESULTS: In the SEER cohort, 65% of reported melanomas were < or = 1.00 mm in thickness and 8.7% were more than 4.00 mm compared with 39% and 10% in the AJCC cohort (P < .001), respectively. AJCC stages were able to discriminate among SEER patient groups with different prognosis. However, SEER survival rates were significantly higher than those in the AJCC study and notably so in patients with T1a lesions (< or = 1 mm without ulceration). This population-specific effect remained significant after controlling for lesion thickness in all substages except stage IIA. CONCLUSION: Although this national population-based study validates the most recent revision of AJCC stages for melanoma, it emphasizes that survival rates are population specific and found them to be generally higher for SEER compared with AJCC patients. Population-specific survival rates should be used in study designs and decisions about patient-specific interventions. PMID- 16258106 TI - Variates of survival in metastatic uveal melanoma. AB - PURPOSE: The course and outcome of metastatic uveal melanoma are not well described. We evaluated the survival of our patients with metastatic uveal melanoma, described factors that correlated with survival, and evaluated the influence of screening tests on time of detection and survival. PATIENTS AND METHODS: All patients with metastatic uveal melanoma seen at Memorial Sloan Kettering Cancer Center between 1994 and 2004 were identified from our database. We recorded date of initial diagnosis, date of metastatic disease, date of last follow-up, site of the first metastasis, how the first metastasis was discovered, treatment, and outcome of therapy. RESULTS: The estimated median survival of the 119 patients analyzed was 12.5 months; 22% of patients were alive at 4 years. Five variates correlated independently with prolonged survival: Lung/soft tissue as only site of first metastasis, treatment with surgery or intrahepatic therapy, female sex, age younger than 60, and a longer interval from initial diagnosis to metastatic disease. Discovering metastatic disease in asymptomatic patients did not correlate with overall survival; 89% of patients had a single organ as the site of first metastasis. Although liver was the most common site, 39.5% of patients had nonliver sites, most commonly lung, as the first site of metastasis. CONCLUSION: A substantial subset of patients with metastatic uveal melanoma survive more than 4 years with metastatic disease. Data on variates of survival and site of first metastasis may guide strategies for screening patients, although our data failed to show a survival advantage in discovering asymptomatic metastatic disease. PMID- 16258107 TI - Opportunities for targeted therapies in hepatocellular carcinoma. AB - Hepatocellular cancer (HCC) is the fifth most common solid tumor worldwide, accounting for 500,000 new cases annually. Although less common in the United States, HCC is expected to increase in incidence over the next two decades largely because of the prevalence of hepatitis C virus infection. A majority of patients present with advanced disease and are not candidates for liver transplantation, surgical resection, or regional therapy. In 60% to 80% of patients with HCC, treatment is complicated by underlying liver cirrhosis and hepatic dysfunction. Systemic treatments are minimally effective, can have significant toxicity, and have not been shown to improve patient survival. New approaches targeting molecular abnormalities specific to HCC are needed to improve patient outcome. This review summarizes the state of knowledge of those key aspects of the molecular pathogenesis of HCC that may represent rational therapeutic targets in this disease. Relevant preclinical and clinical information on novel compounds directed toward abnormalities in HCC is reviewed. PMID- 16258108 TI - The bereaved parent. PMID- 16258109 TI - Variable problems in lymphomas: CASE 1. Burkitt's lymphoma presenting with central airway obstruction. PMID- 16258110 TI - Variable problems in lymphomas: CASE 2. Sarcoidosis mimicking progressive lymphoma. PMID- 16258111 TI - Variable problems in lymphomas: CASE 3. Spontaneous regression of HIV-associated Burkitt's lymphoma of the cecum. PMID- 16258112 TI - Epidermal growth factor receptor expression in breast cancer. PMID- 16258113 TI - Therapy-related myelodysplastic syndrome and acute myeloid leukemia after autologous bone marrow transplantation: dosis facit venenum? PMID- 16258114 TI - Analysis of outcome by response flawed. PMID- 16258115 TI - Randomized phase II trials and prostate-specific antigen endpoints in prostate cancer: much ado about nothing? PMID- 16258116 TI - Chronic lymphocytic leukemia: to transplant or not to transplant... that is the question? PMID- 16258117 TI - Palliation in incurable head and neck cancers: chemotherapy? PMID- 16258118 TI - Is pathologic complete response a valid surrogate parameter of treatment efficacy in HER2 positive breast cancer patients undergoing primary chemotherapy plus trastuzamab? PMID- 16258119 TI - Patients with HIV with Burkitt's lymphoma have a worse outcome than those with diffuse large-cell lymphoma also in the highly active antiretroviral therapy era. PMID- 16258120 TI - Incidence of cancer in individuals receiving chronic zopiclone or eszopiclone requires prospective study. PMID- 16258121 TI - Surrogate markers for antiangiogenic therapy and dose-limiting toxicities for bevacizumab with radiation and chemotherapy: continued experience of a phase I trial in rectal cancer patients. PMID- 16258122 TI - Comprehensive review of chemotherapy in patients with metastatic breast cancer. PMID- 16258123 TI - Inter-religious perspectives on hope and limits in cancer treatment: one Buddhist chaplain's response to the case. PMID- 16258125 TI - EU takes further steps to reduce the risk from avian influenza. PMID- 16258129 TI - Bringing paraprofessionals into the VSA. PMID- 16258130 TI - Living on borrowed time? Meeting the demands of 24-hour cover. PMID- 16258133 TI - Virological and serological evidence of bovine herpesvirus type 4 in cattle in Northern Ireland. AB - Bovine herpesvirus type 4 (BHV-4), a member of the genus Rhadinovirus, subfamily Gammaherpesvirinae, within the family Herpesviridae, was isolated in fetal bovine lung cells from samples of vaginal discharge taken from a dairy herd in which approximately 50 per cent of the cattle developed metritis after calving. The identity of the isolate was confirmed by immunofluorescent staining with a BHV-4 specific monoclonal antibody and partial sequencing of a portion of the glycoprotein B gene. Serological testing failed to demonstrate a significant association between the exposure of the cattle to BHV-4 and the metritis, but several cattle seroconverted during the periparturient period, consistent with the recrudescence and shedding of virus associated with the stresses of parturition and the onset of lactation. Despite the previous failure to detect BHV-4 in Northern Ireland, a serological survey of 999 cattle in 49 dairy herds and 51 beef herds found widespread evidence of exposure: 29 of the dairy herds and 35 of the beef herds contained one or more seropositive cattle, and 33.3 per cent of the dairy cattle and 23.3 per cent of the beef cattle were positive. PMID- 16258134 TI - Ultrasonographic assessment and ultrasound-guided biopsy of the retropharyngeal lymph nodes in Asian elephants (Elephas maximus). AB - Endotheliotropic herpesvirus causes a fatal disease in young Asian elephants, but there are no methods for identifying latent carriers of the virus. During the postmortem study of one female African elephant and three male and two female Asian elephants, a lymph node located bilaterally caudoventral to the parotid gland, approximately 1.5 to 5 cm below the skin, was identified as suitable for transcutaneous ultrasound-guided biopsy. An ultrasonographic assessment and two biopsies were performed on 39 Asian elephants, and these lymph nodes were classified ultrasonographically as active, inactive or chronically active. The calculated mean (se) volume of 10 active lymph nodes was 17.4 (6.9) cm(3), and that of three chronically active lymph nodes was 10.6 (1.0) cm(3), whereas the mean volume of 17 inactive lymph nodes was 3.1 (0.6) cm(3). The presence of lymph node tissue in samples obtained by ultrasound-guided biopsy from three animals that were maintained under conditions that allowed for additional sampling was confirmed histologically. The dna extracted from the lymphoid tissue and the whole blood of all the elephants was negative for endotheliotropic herpesvirus by PCR. PMID- 16258135 TI - Variations in the concentration of zinc in the blood of Icelandic horses. AB - The effect of factors including the horses' farm environment, their sex and age and whether they suffered from summer seasonal recurrent dermatitis (sweet itch) on the concentrations of zinc in the plasma, whole blood and blood cells of 104 Icelandic horses was investigated. Its concentration in plasma varied significantly between farms (P<0.01), but its concentration in blood and blood cells was not influenced by any of the variables. The concentration of zinc in the blood cells was 10.5 times greater than in plasma, but its concentration in plasma was not correlated with its concentration in whole blood or blood cells owing to the variability in the proportion of whole blood zinc present in plasma (relative plasma zinc), which ranged between 9 and 24 per cent. This variability was significantly influenced by a three-way interaction between farm, sex and sweet itch (P<0.05). Relative plasma zinc was positively correlated with absolute plasma zinc (r=0.78, P<0.001) and negatively correlated with whole blood and blood cellular zinc (r=-0.58, r=-0.71, P<0.001). PMID- 16258136 TI - Evaluation of an ELISA to detect antibodies to maedi-visna virus in individual and pooled samples of milk from sheep. AB - An elisa was used to detect antibodies to maedi-visna virus in samples of serum and milk from individual sheep; the results obtained indicated that the elisa can be used to detect antibodies in milk. The assay was also applied to samples of bulk-tank milk; a standard curve was created and used to calculate the seroprevalence of maedi-visna in 11 flocks of sheep and the results were compared with the results obtained by applying the elisa to individual serum samples. There was good agreement between the seroprevalences calculated from the standard curve for bulk-tank milk and from the individual serum samples. PMID- 16258137 TI - Change in the prevalence of Thelazia species in bovine eyes in England. PMID- 16258138 TI - Seminoma in a southern white rhinoceros (Ceratotherium simum simum). PMID- 16258139 TI - Uterine horn torsion in a pregnant cat. PMID- 16258140 TI - Individual quarter agalactia in dairy cows. PMID- 16258141 TI - Action on bovine TB. PMID- 16258143 TI - Changing pattern of mastitis isolates. PMID- 16258144 TI - Management of fear of fireworks in dogs. PMID- 16258145 TI - Circulating human heat shock protein 60 in the blood of healthy teenagers: a novel determinant of endothelial dysfunction and early vascular injury? PMID- 16258146 TI - Inflammation warms up the metabolic syndrome. PMID- 16258147 TI - Iron chelation and vascular function: in search of the mechanisms. PMID- 16258148 TI - Beyond blood pressure: subtle effects of drug classes. PMID- 16258149 TI - Ethnic differences in arterial responses, inflammation, and metabolic profiles: possible insights into ethnic differences in cardiovascular disease and stroke. PMID- 16258150 TI - Metabolic syndrome scientific statement by the American Heart Association and the National Heart, Lung, and Blood Institute. PMID- 16258151 TI - ATVB in focus: novel mediators and mechanisms in angiogenesis and vasculogenesis. PMID- 16258152 TI - Molecular diagnosis of severe acute respiratory syndrome: the state of the art. AB - Severe acute respiratory syndrome (SARS) first appeared in Guangdong Province, China, in November 2002. Although virus isolation and serology were useful early in the SARS outbreak for diagnosing new cases, these tests are not generally useful because virus culture requires a BSL-3 laboratory and seroconversion is often delayed until 2 to 3 weeks after infection. The first qualitative reverse transcriptase-polymerase chain reaction tests for SARS-coronavirus (CoV) were sensitive and capable of detecting 1 to 10 genome equivalents. These assays were quickly supplemented with quantitative real-time assays that helped elucidate the natural history of SARS, particularly the initial presence of low viral loads in the upper respiratory tract and high viral loads in the lower respiratory tract. The unique natural history of SARS-CoV infection dictates the testing of both respiratory and nonrespiratory specimens, the testing of multiple specimens from the same patient, and sending out positives to be confirmed by a reference laboratory. Commercially available reverse transcriptase-polymerase chain reaction tests for SARS have recently appeared; however, meaningful evaluations of these assays have not yet been performed and their true performance has not been determined. These and other issues related to diagnosis of SARS-CoV infection are discussed in this review. PMID- 16258153 TI - A practical approach to the detection of prognostically significant genomic aberrations in multiple myeloma. AB - Multiple myeloma (MM) is a malignancy of differentiated B lymphocytes and has remained an incurable disease. Chromosomal abnormalities are among the most important prognostic parameters for MM. Cytoplasm immunoglobulin-enhanced interphase fluorescent in situ hybridization (FISH) has been a standard cell targeting method for identifying genomic aberrations in MM. We have developed another cell-targeting approach by using CD138 magnetic microbeads to sort plasma cells for FISH analysis. The FISH panel consisted of four probes targeting RB-1, D13S319, immunoglobulin H, and p53 loci. We reviewed the FISH and conventional cytogenetic results of 60 patients with MM. The present cell-targeting approach in conjunction with the FISH probe panel was more sensitive than FISH performed on untargeted cells in detecting prognostically significant genomic aberrations (72 versus 24%, P = 0.0016). The frequencies of genomic abnormalities identified were similar to previously reported data obtained with the standard cell targeting method. Therefore, our cell-targeting approach and FISH panel reliably detect prognostically important genomic abnormalities in MM and are potentially suitable for widespread use. PMID- 16258154 TI - Immunoglobulin mutational status detected through single-round amplification of partial V(H) region represents a good prognostic marker for clinical outcome in chronic lymphocytic leukemia. AB - The immunoglobulin (Ig) mutational status in B-cell chronic lymphocytic leukemia (CLL) distinguishes two subsets of patients with different prognosis. Ig status detection is commonly performed with a panel of V(H) family-specific primers. Although this method detects clonal VDJ rearrangement in virtually all cases, it is technically cumbersome and therefore not widely used clinically. Here, we describe a simple and rapid method to establish the mutational status of IgV(H) in CLL. The method is based on a consensus V(H) FR2 primer, used in both polymerase chain reaction (PCR) and sequencing reactions. Overall, monoclonal B cell populations were detected in 163 of 189 CLL patients (86%). The prognostic value of IgV(H) mutational status was then evaluated by analyzing survival in 146 CLL cases using different V(H) homology cutoffs. CLL prognostic groups were best separated by the classical 98% cutoff: median survival was 127 and 206 months in unmutated and mutated CLL cases, respectively (P = 0.0023). V(H) FR2 consensus and V(H) family PCR were compared in 41 cases, correctly assigning all cases by both methods. Therefore, we suggest a sequential strategy to detect immunoglobulin mutational status in CLL patients by first using the approach described in this study followed by alternative V(H) family-specific PCRs for negative cases. PMID- 16258155 TI - Real-time polymerase chain reaction for detecting bacterial DNA directly from blood of neonates being evaluated for sepsis. AB - Speed is of the essence when evaluating an infant with symptoms consistent with sepsis. Because of the high morbidity and mortality associated with neonatal sepsis, infants receive multiple, broad-spectrum antibiotics before receiving finalized blood culture results. Incorporating an additional, reliable, yet rapid assay to detect bacteria directly from blood would facilitate timely diagnosis and appropriate care. To this end, we designed a real-time polymerase chain reaction (PCR) assay targeting the highly conserved 380 bases of 16S rDNA. DNA was extracted from whole-blood samples using a Qiagen column. The limit of detection for the TaqMan-based assay, using a Smartcycler instrument, was 40, 50, or 2000 colony-forming units per milliliter of blood (CFU/ml) of Escherichia coli, group B Streptococcus, and Listeria monocytogenes, respectively, when white blood cell counts were below 39,000/microl. Implementing this approach requires less than 4 hours for both sample preparation and real-time PCR compared with 1 to 2 days to detect growth in culture or 5 days to finalize no-growth culture results. There was an overall agreement between the results of culture and real time PCR of 94.1% (80 of 85) in this study. These results suggest that molecular techniques can augment culture-based methods for diagnosing neonatal sepsis, especially in infants whose mothers have received intrapartum antibiotic prophylaxis. PMID- 16258156 TI - Application of BIOMED-2 primers in fixed and decalcified bone marrow biopsies: analysis of immunoglobulin H receptor rearrangements in B-cell non-Hodgkin's lymphomas. AB - The detection of clonality in lymphomas was recently improved by the BIOMED-2 approach, but analysis of fixed tissues is limited. Here, we adapted the BIOMED-2 protocol for examining immunoglobulin H (IgH) receptor rearrangements in fixed, decalcified bone marrow biopsies (BMBs) for clonality analysis in B-cell non Hodgkin's lymphomas (B-NHL). The study included 111 decalcified BMBs (12 formalin fixed and 99 glutardialdehyde fixed), with B-NHL (n = 85), T-NHL (n = 8), or reactive infiltrates (n = 18). Initially, IgH FRIII polymerase chain reaction (PCR) analysis of crude DNA extracts from 75 glutardialdehyde-fixed BMBs (B-NHLs) using a standard seminested PCR resulted in clonal peaks in 46 of 75 (61.3%) BMBs compared with 19 of 70 (27.1%) for the original BIOMED-2 protocol. Modifications to both DNA extraction and PCR reaction improved the detection rate to 26 of 36 (72.2%) for BIOMED-2 primers, including 10 of 15 (66.7%) cases not detected by our standard IgH analysis. Moreover, introducing the same modifications for analysis of the FRII region by BIOMED-2 primers revealed clonal peaks in 19 of 36 (52.8%) B-NHLs compared with 5 of 70 (7.1%) for the original BIOMED-2 protocol. Together, analysis of FRII and FRIII regions by the modified BIOMED-2 protocol increased the detection rate to 31 of 36 (86.1%), particularly for BMBs with histological evidence of follicular lymphoma (FRIII, 70%; FRII, 90%). In summary, this study provides an improved protocol for detection of clonality by IgH specific BIOMED-2 primers in fixed, decalcified bone marrow biopsies. PMID- 16258157 TI - Simultaneous genotyping of DRB1/3/4/5 loci by oligonucleotide microarray. AB - Matching of the HLA antigens for donor-recipient in transplantation, disease predisposition or protection, population studies, and forensic testing requires accurate but simple typing methods. Here, we describe a DNA-based tissue-typing assay that determines the haplotype of the DRB1/3/4/5 loci in hy-bridization of oligonucleotide array after sample amplification. Using this multianalyte DNA hybridization system, we analyzed seven regions of exon 2 of DRB loci that have single-base discrimination. Thirty-six oligonucleotide probes complementary to the alleles of interest were immobilized on each microslide. The efficiency and specificity of identifying DRB genotypes using the oligonucleotide arrays was evaluated by blinded analysis of 147 samples from reference standards and subjects. The established method provides a rapid and inexpensive DRB "low resolution" typing tool for prescreening a large number of samples. PMID- 16258158 TI - FokI polymorphism, vitamin D receptor, and interleukin-1 receptor haplotypes are associated with type 1 diabetes in the Dalmatian population. AB - Vitamin D and interleukin (IL)-1 have been suggested to function in the pathogenesis of type 1 diabetes mellitus (T1DM). Therefore, we examined the influence of gene polymorphisms in vitamin D receptor (VDR) and interleukin-1 receptor type I (IL-1-R1) on susceptibility to T1DM in the Dalmatian population of South Croatia. We genotyped 134 children with T1DM and 132 controls; for FokI polymorphism studies, we extended the control group to an additional 102 patients. The VDR gene polymorphism FokI displayed unequal distribution (P = 0.0049) between T1DM and control groups, with the ff genotype occurring more frequently in T1DM individuals whereas the VDR gene polymorphism Tru9I did not differ in frequency between studied groups. All tested polymorphisms of the IL-1 R1 gene [PstI, HinfI, and AluI (promoter region) and PstI-e (exon 1B region)] displayed no differences between cases and controls. Haplotype analysis of the VDR gene (FokI, BsmI, ApaI, TaqI, Tru9I) and of the IL-1-R1 gene (PstI, HinfI, AluI, PstI-e) found haplotypes VDR FbATu (P = 0.0388) and IL-1-R1 phap' (P = 0.0419) to be more frequent in T1DM patients whereas the BatU haplotype occurred more often in controls (P = 0.0064). Our findings indicate that the VDR FokI polymorphism and several VDR and IL-1-R1 haplotypes are associated with susceptibility to T1DM in the Dalmatian population. PMID- 16258159 TI - An enhanced polymerase chain reaction assay to detect pre- and full mutation alleles of the fragile X mental retardation 1 gene. AB - Several diagnostic strategies have been applied to the detection of FMR1 gene repeat expansions in fragile X syndrome. Here, we report a novel polymerase chain reaction-based strategy using the Expand Long Template PCR System (Roche Diagnostics, Mannheim, Germany) and the osmolyte betaine. Repeat expansions up to approximately 330 CGGs in males and up to at least approximately 160 CGGs in carrier women could be easily visualized on ethidium bromide agarose gels. We also demonstrated that fluorescence analysis of polymerase chain reaction products was a reliable tool to verify the presence of premutation and full mutation alleles both in males and in females. This technique, primarily designed to detect premutation alleles, can be used as a routine first screen for expanded FMR1 alleles. PMID- 16258160 TI - Simultaneous detection and quantification of mitochondrial DNA deletion(s), depletion, and over-replication in patients with mitochondrial disease. AB - Heterogeneous clinical expression of mitochondrial DNA (mtDNA) disorders depends on both qualitative and quantitative changes in mtDNA. We developed a sensitive and effective method that simultaneously detects mtDNA deletion(s) and quantifies total mtDNA content. The percentage of deletions and mtDNA content of 19 patients with single or multiple deletions were analyzed by real-time quantitative polymerase chain reaction (real-time qPCR) using TaqMan probes specific for mtDNA (tRNA leu(UUR), ND4, ATPase8, and D-loop regions) and nuclear DNA (AIB1, beta-2 microglobulin, and beta-actin). The proportion of deletion mutants determined by real-time qPCR was consistent with that determined by Southern analysis. Most patients with mtDNA deletions also demonstrated compensatory mtDNA over replication. Multiple mtDNA deletions that were not detectable by Southern analysis due to low percentage of each deletion molecule were readily detected and quantified by real-time qPCR. Furthermore, 12 patients with clinical features and abnormal biochemical/histopathological results consistent with mitochondrial respiratory chain disorders without identified mtDNA mutations had either substantially depleted or significantly over-replicated mtDNA content, supporting the diagnosis of mitochondrial disease. Our results demonstrate that both qualitative and quantitative analyses are important in molecular diagnosis of mitochondrial diseases. The presence of deletion(s) and mtDNA depletion or compensatory over-replication can be determined simultaneously by real-time qPCR. PMID- 16258161 TI - Mass spectrometry-based loss of heterozygosity analysis of single-nucleotide polymorphism loci in paraffin embedded tumors using the MassEXTEND assay: single nucleotide polymorphism loss of heterozygosity analysis of the protein tyrosine phosphatase receptor type J in familial colorectal cancer. AB - As the number of identified single-nucleotide polymorphisms (SNPs) increases, high-throughput methods are required to characterize the informative loci in large patient series. We investigated the feasibility of MassEXTEND LOH analysis using Sequenom's MassArray RT software, a mass spectrometry method, as an alternative to determine loss of heterozygosity (LOH). For this purpose, we studied the c.827A>C SNP (1176A>C p.Gln276Pro) in protein tyrosine phosphatase receptor type-J (PTPRJ), which is frequently deleted in human cancers. In sporadic colorectal cancer (CRC), c.827A>C showed allele-specific LOH of the c.827A allele, which is important because LOH of PTPRJ may be an early event during sporadic CRC. To elucidate the impact of this low-penetrance gene on familial CRC, we studied c.827A>C in 222 familial CRC cases and 156 controls. In 6.2% of the A/C genotyped CRC samples, LOH of c.827A was observed with MassEXTEND LOH analysis and confirmed by conventional sequencing. Furthermore, a case with LOH of c.827A showed no LOH in 22 synchronously detected adenomas, including one with malignant transformation. The importance of the PTPRJ- c.827A>C SNP appears to be limited in familial CRC. We conclude that MassEXTEND LOH analysis (using Sequenom's MassARRAY RT software) is a sensitive, high-throughput, and cost effective method to screen SNP loci for LOH in formalin-fixed paraffin-embedded tissue. PMID- 16258162 TI - Critical evaluation of real-time reverse transcriptase-polymerase chain reaction for the quantitative detection of cytokeratin 20 mRNA in colorectal cancer patients. AB - We evaluated the usefulness of cytokeratin 20 (CK20) mRNA expression in the quantitative detection of circulating tumor cells in the blood of patients with colorectal cancer (CRC). Blood samples from healthy volunteers (HVs; n = 37), patients with localized (n = 42) and metastatic colorectal cancer (n = 40), and patients with chronic inflammatory bowel disease (CID; n = 15) were examined. After immunomagnetic enrichment using microbeads against human epithelial antigen, total RNA was extracted, reverse transcribed, and analyzed by real-time reverse transcriptase-polymerase chain reaction using the LightCycler instrument. CK20 expression in peripheral blood was found in 46 of 82 (56%) patients with CRC, 8 of 37 (22%) HVs, and 9 of 15 (60%) patients with CID. Levels of CK20 mRNA were significantly higher in blood samples from CRC patients (median 681) than in blood samples from HVs (median 0) (P = 0.001), whereas no difference could be detected between patients with CRC and CID. Although the present technique could not distinguish CRC from CID, the method warrants further efforts to improve sample preparation and tumor cell enrichment, which may render real-time CK20 reverse transcriptase-polymerase chain reaction a feasible technique in identifying circulating tumor cells in peripheral blood of cancer patients. PMID- 16258163 TI - Sensitive detection of polyalanine expansions in PHOX2B by polymerase chain reaction using bisulfite-converted DNA. AB - Congenital central hypoventilation syndrome, also known as Ondine's curse, is characterized by idiopathic abnormal control of respiration during sleep. Recent studies indicate that a polyalanine expansion of PHOX2B is relevant to the pathogenesis of this disorder. However, it is difficult to detect the repeated tract because its high GC content inhibits conventional polymerase chain reaction (PCR) amplification. Here, we describe a bisulfite treatment for DNA in which uracil is obtained by deamination of unmethylated cytosine residues. Deamination of DNA permitted direct PCR amplification that yielded a product of 123 bp for the common 20-residue repetitive tract with replacement of C with T by sequencing. It settled allele dropouts accompanied by insufficient amplification of expanded alleles. The defined procedure dramatically improved detection of expansions to 9 of 10 congenital central hypoventilation syndrome patients examined in a previous study. The chemical conversion of DNA before PCR amplification facilitates effective detection of GC-rich polyalanine tracts. PMID- 16258164 TI - Molecular diagnosis of necrotizing fasciitis by 16S rRNA gene sequencing and superantigen gene detection. AB - We report the use of molecular techniques in the diagnosis of a case of culture negative necrotizing fasciitis occurring in a 32-year-old female with no significant past medical history and who died within 36 hours of admission. Paraffin-embedded tissue sections from the popliteal fossa region obtained at autopsy showed hemorrhage, necrosis, and mild inflammation by hematoxylin and eosin staining. Tissue gram stain showed numerous gram-positive organisms arranged in clusters. The sequences of the first 500 bp of bacterial 16S rRNA gene amplified from the lesion were identical to a Lancefield group A beta hemolytic Streptococcus pyogenes. Streptococcal pyrogenic exotoxin A and B superantigen genes were detected and an emm type 1 was determined by polymerase chain reaction and sequencing from the lesion. This confirmed the etiology of the patient's rapid deterioration with multisystem organ failure. PMID- 16258165 TI - The effect of high-dose simvastatin on triglyceride-rich lipoprotein metabolism in patients with type 2 diabetes mellitus. AB - Statins decrease triglycerides (TGs) in addition to decreasing low density lipoprotein-cholesterol. Although the mechanism for the latter effect is well understood, it is still unclear how TG decrease is achieved with statin therapy. Because hypertriglyceridemia is common in obese patients with type 2 diabetes mellitus, we studied triglyceride-rich lipoprotein triglyceride (TRL-TG) turnover in 12 such subjects using stable isotopically labeled glycerol. The diabetic subjects were studied after 12 weeks of placebo and after a similar course of therapy with simvastatin (80 mg daily) in a single-blind design. The results were compared with those from six nonobese nondiabetic control subjects. Simvastatin therapy reduced serum TGs by 35% in the diabetic subjects. Compared with the control subjects, TRL-TG secretion was almost 2-fold higher in the diabetic subjects (45.4 +/- 4.9 vs. 24.4 +/- 1.9 micromol/min; P < 0.002) and was unaffected by simvastatin therapy. However, TRL-TG clearance was significantly increased in the diabetic subjects during simvastatin treatment compared with placebo (0.25 +/- 0.03 vs. 0.16 +/- 0.02 pools/h; P < 0.002). This change was accompanied by a 49% increase in preheparin plasma lipase activity (P < 0.03) and a 21% increase in postheparin LPL activity (P < 0.01). Together, these findings provide strong evidence that the effect of statins on serum TGs is related to an increase in LPL activity, resulting in accelerated delipidation of TRL particles. The effect of high-dose simvastatin on triglyceride-rich lipoprotein metabolism in patients with type 2 diabetes mellitus. PMID- 16258166 TI - Apolipoprotein A-V: a potential modulator of plasma triglyceride levels in Turks. AB - The apolipoprotein A-V gene (APOA5) plays an important role in determining plasma triglyceride levels. We studied the effects of APOA5 polymorphisms on plasma triglyceride levels in Turks, a population with low levels of HDL cholesterol and a high prevalence of coronary artery disease. We found 15 polymorphisms, three of which were novel. Seven haplotype-tagging single nucleotide polymorphisms (SNPs) were chosen and genotyped in approximately 3,000 subjects. The rare alleles of the -1464T>C, -1131T>C, S19W, and 1259T>C SNPs were significantly associated with increased triglyceride levels (19-86 mg/dl; P < 0.05) and had clear gene-dose effects. Haplotype analysis of the nine common APOA5 haplotypes revealed significant effects on triglyceride levels (P < 0.001). Detailed analysis of haplotypes clearly showed that the -1464T>C polymorphism had no effect by itself but was a marker for the -1131T>C, S19W, and 1259T>C polymorphisms. The -1131T>C and 1259T>C polymorphisms were in a strong but incomplete linkage disequilibrium and appeared to have independent effects. Thus, the APOA5 -1131T>C, S19W, and 1259T>C rare alleles were associated with significant increases in plasma triglyceride levels. At least one of these alleles was present in approximately 40% of the Turks. Similar associations were observed for -1131T>C and S19W in white Americans living in San Francisco, California. PMID- 16258167 TI - A membrane defect in the pathogenesis of the Smith-Lemli-Opitz syndrome. AB - The Smith-Lemli-Opitz syndrome (SLOS) is an often lethal birth defect resulting from mutations in the gene responsible for the synthesis of the enzyme 3beta hydroxy-steroid-Delta7-reductase, which catalyzes the reduction of the double bond at carbon 7 on 7-dehydrocholesterol (7-DHC) to form unesterified cholesterol. We hypothesize that the deficiency in cholesterol biosynthesis and subsequent accumulation of 7-DHC in the cell membrane leads to defective composition, organization, dynamics, and function of the cell membrane. Using skin fibroblasts obtained from SLOS patients, we demonstrate that the SLOS membrane has increased 7-DHC and reduced cholesterol content and abnormal membrane fluidity. X-ray diffraction analyses of synthetic membranes prepared to mimic SLOS membranes revealed atypical membrane organization. In addition, calcium permeability is markedly augmented, whereas membrane-bound Na+/K+ATPase activity, folate uptake, inositol-1,4,5-trisphosphate signaling, and cell proliferation rates are markedly suppressed. These data indicate that the disturbance in membrane sterol content in SLOS, likely at the level of membrane caveolae, directly contributes to the widespread tissue abnormalities in this disease. PMID- 16258168 TI - Integrating cytomics and proteomics. AB - Systems biology along with what is now classified as cytomics provides an excellent opportunity for cytometry to become integrated into studies where identification of functional proteins in complex cellular mixtures is desired. The combination of cell sorting with rapid protein-profiling platforms offers an automated and rapid technique for greater clarity, accuracy, and efficiency in identification of protein expression differences in mixed cell populations. The integration of cell sorting to purify cell populations opens up a new area for proteomic analysis. This article outlines an approach in which well defined cell analysis and separation tools are integrated into the proteomic programs within a core laboratory. In addition we introduce the concepts of flow cytometry sorting to demonstrate the importance of being able to use flow cytometry as a cell separation technology to identify and collect purified cell populations. Data demonstrating the speed and versatility of this combination of flow cytometry based cell separation and protein separation and subsequent analysis, examples of protein maps from purified sorted cells, and an analysis of the overall procedure will be shown. It is clear that the power of cell sorting to separate heterogeneous populations of cells using specific phenotypic characteristics increases the power of rapid automated protein separation technologies. PMID- 16258169 TI - Decorin core protein secretion is regulated by N-linked oligosaccharide and glycosaminoglycan additions. AB - Expression of decorin using the vaccinia virus/T7 expression system resulted in secretion of two distinct glycoforms: a proteoglycan substituted with a single chondroitin sulfate chain and N-linked oligosaccharides and a core protein glycoform substituted with N-linked glycans but without a glycosaminoglycan chain. In this report, we have addressed two distinct questions. What is the rate limiting step in glycosaminoglycan synthesis? Is glycosylation with either N linked oligosaccharides or glycosaminoglycan required for secretion of decorin? N terminal sequencing of the core protein glycoform, the addition of benzyl-beta-d xyloside, and a UDP-xylose: core protein beta-d-xylosyltransferase activity assay show that xylosylation is a rate-limiting step in chondroitin sulfate biosynthesis. Decorin can be efficiently secreted with N-linked oligosaccharides alone or with a single chondroitin sulfate chain alone; however, there is severely impaired secretion of core protein devoid of any glycosylation. A decorin core protein mutant devoid of N-linked oligosaccharide attachment sites will not be secreted by Chinese hamster ovary cells deficient in xylosyltransferase or by parental Chinese hamster ovary wild type cells if the xylosyltransferase recognition sequence is disrupted. This finding suggests that quality control mechanisms sensitive to an absence of N-linked oligosaccharides can be abrogated by interaction of the core protein with the glycosaminoglycan synthetic machinery. We propose a model of regulation of decorin secretion that has several components, including appropriate substitution with N-linked oligosaccharides and factors involved in glycosaminoglycan synthesis. PMID- 16258170 TI - VEGF165-binding sites within heparan sulfate encompass two highly sulfated domains and can be liberated by K5 lyase. AB - The vascular endothelial growth factor (VEGF) family of proteins controls the formation and growth of blood vessels. The most potent and widely expressed isoform, VEGF165, is secreted as a disulfide-linked homodimer with two identical heparin-binding sites. Interactions with heparan sulfate (HS) regulate the diffusion, half-life, and affinity of VEGF165 for its signaling receptors. We have determined a number of key HS structural features that mediate the specific binding of the VEGF165 dimer. Carboxylate groups and 2-O-, 6-O-, and N-sulfation of HS contributed to the strength of the VEGF165 interaction; however, 6-O sulfates appeared to be particularly important. Cleavage of HS by heparinase, heparitinase, or heparanase severely reduced VEGF165 binding. In contrast, K5 lyase-cleaved HS retained significant VEGF165 affinity, suggesting that binding sites for the growth factor are present within extended stretches of sulfation. Binding studies and molecular modeling demonstrated that an oligosaccharide 6 or 7 residues long was sufficient to fully occupy the heparin-binding site of a VEGF165 monomer. The data presented are consistent with a model whereby the two heparin-binding sites of the VEGF165 dimer interact simultaneously with highly sulfated S-domain regions of the HS chain that can be linked through a stretch of transition sequence. PMID- 16258171 TI - The related retinoblastoma (pRb) and p130 proteins cooperate to regulate homeostasis in the intestinal epithelium. AB - pRb, p107, and p130 are related proteins that play a central role in the regulation of cell cycle progression and terminal differentiation in mammalian cells. Nevertheless, it is still largely unclear how these proteins achieve this regulation in vivo. The intestinal epithelium is an ideal in vivo system in which to study the molecular pathways that regulate proliferation and differentiation because it exists in a constant state of development throughout an animal's lifetime. We studied the phenotypic effects on the intestinal epithelium of mutating Rb and p107 or p130. Although mutating these genes singly had little or no effect, loss of pRb and p107 or p130 together produced chronic hyperplasia and dysplasia of the small intestinal and colonic epithelium. In Rb/p130 double mutants this hyperplasia was associated with defects in terminal differentiation of specific cell types and was dependent on the increased proliferation seen in the epithelium of mutant animals. At the molecular level, dysregulation of the Rb pathway led to an increase in the expression of Math1, Cdx1, Cdx2, transcription factors that regulate proliferation and differentiation in the intestinal epithelium. The absence of Cdx1 function in Rb/p130 double mutant mice partially reverted the histologic phenotype by suppressing ectopic mitosis in the epithelium. These studies implicate the Rb pathway as a regulator of epithelial homeostasis in the intestine. PMID- 16258172 TI - High affinity binding between copper and full-length prion protein identified by two different techniques. AB - The cellular prion protein is known to be a copper-binding protein. Despite the wide range of studies on the copper binding of PrP, there have been no studies to determine the affinity of the protein on both full-length prion protein and under physiological conditions. We have used two techniques, isothermal titration calorimetry and competitive metal capture analysis, to determine the affinity of copper for wild type mouse PrP and a series of mutants. High affinity copper binding by wild type PrP has been confirmed by the independent techniques indicating the presence of specific tight copper binding sites up to femtomolar affinity. Altogether, four high affinity binding sites of between femto- and nanomolar affinities are located within the octameric repeat region of the protein at physiological pH. A fifth copper binding site of lower affinity than those of the octameric repeat region has been detected in full-length protein. Binding to this site is modulated by the histidine at residue 111. Removal of the octameric repeats leads to the enhancement of affinity of this fifth site and a second binding site outside of the repeat region undetected in the wild type protein. High affinity copper binding allows PrP to compete effectively for copper in the extracellular milieu. The copper binding affinities of PrP have been compared with those of proteins of known function and are of magnitudes compatible with an extracellular copper buffer or an enzymatic function such as superoxide dismutase like activity. PMID- 16258173 TI - Adenovirus-mediated gene transfer of mutated IkappaB kinase and IkappaBalpha reveal NF-kappaB-dependent as well as NF-kappaB-independent pathways of HAS1 activation. AB - It has become increasingly clear that hyaluronan is more than the simple matrix molecule it was once thought to be but instead takes part in a multitude of biological functions. Three genes encode for hyaluronan synthases (HAS). We demonstrated earlier that HAS2 and HAS3 are constitutively activated in type-B synoviocytes (fibroblast-like synoviocytes) and, furthermore, that the only gene that readily responds to stimulation with a series of proinflammatory cytokines is HAS1. Here we probe the involvement of the transcription factor NF-kappaB in induced and noninduced HAS activation. Transforming growth factor (TGF) beta1 as well as interleukin (IL)-1beta are both strong inducers of HAS1 transcription. Stimulation of fibroblast-like synoviocytes with IL-1beta resulted in rapid degradation of IkappaBalpha, an event that was preceded by IkappaBalpha phosphorylation. Interestingly, TGFbeta1 neither affected IkappaBalpha levels, nor did it cause phosphorylation of IkappaBalpha. In addition, TGFbeta1 had no effect on IkappaBbeta and IkappaBepsilon levels. Electrophorectic mobility shift assays demonstrate that IL-1beta is a potent inducer of NF-kappaB translocation; however, TGFbeta1 treatment did not result in shifting bands. Two adenovirus constructs were used to further clarify differences in TGFbeta1- and IL-1beta induced HAS1 activation. Overexpressing IkappaBalpha completely abolished the IL 1beta effect on HAS1 but did not interfere with TGFbeta1-induced HAS1 mRNA accumulation. Identical results were obtained when a dominant negative IKK was overexpressed. Interestingly, neither overexpression of IkappaBalpha nor of IKK had any effect on HAS2 and HAS3 mRNA levels. Taken together, HAS1 can be activated by distinct pathways; IL-1beta utilizes NF-kappaB, and TGFbeta1 does not. Furthermore, HAS2 and HAS3 are activated without the involvement of NF kappaB. PMID- 16258174 TI - Functional analysis of the bacteriophage T4 DNA-packaging ATPase motor. AB - Packaging of double-stranded DNA into bacteriophage capsids is driven by one of the most powerful force-generating motors reported to date. The phage T4 motor is constituted by gene product 16 (gp16) (18 kDa; small terminase), gp17 (70 kDa; large terminase), and gp20 (61 kDa; dodecameric portal). Extensive sequence alignments revealed that numerous phage and viral large terminases encode a common Walker-B motif in the N-terminal ATPase domain. The gp17 motif consists of a highly conserved aspartate (Asp255) preceded by four hydrophobic residues (251MIYI254), which are predicted to form a beta-strand. Combinatorial mutagenesis demonstrated that mutations that compromised hydrophobicity, or integrity of the beta-strand, resulted in a null phenotype, whereas certain changes in hydrophobicity resulted in cs/ts phenotypes. No substitutions, including a highly conservative glutamate, are tolerated at the conserved aspartate. Biochemical analyses revealed that the Asp255 mutants showed no detectable in vitro DNA packaging activity. The purified D255E, D255N, D255T, D255V, and D255E/E256D mutant proteins exhibited defective ATP binding and very low or no gp16-stimulated ATPase activity. The nuclease activity of gp17 is, however, retained, albeit at a greatly reduced level. These data define the N terminal ATPase center in terminases and show for the first time that subtle defects in the ATP-Mg complex formation at this center lead to a profound loss of phage DNA packaging. PMID- 16258175 TI - Potent inhibition of carcinogen-bioactivating cytochrome P450 1B1 by the p53 inhibitor pifithrin alpha. AB - Pifithrin alpha (PFTalpha) is a chemical compound that inhibits p53-mediated gene activation and apoptosis. It has also been recently shown to alter metabolism of carcinogenic polycyclic aromatic hydrocarbons (PAHs). This has led us to examine the effect of PFTalpha on the activity of cytochrome P-450 (CYP) 1 isoforms, known to metabolize PAHs, such as benzo(a)pyrene (BP), into mutagenic metabolites. We report that PFTalpha caused a potent inhibition of CYP1-related activity as measured by ethoxyresorufin O-deethylase activity in CYP1-containing MCF-7 cells and liver microsomes. It also directly affected the catalytic activity of human recombinant CYP1A1, CYP1A2 and CYP1B1 isoforms, with a potent inhibitory effect towards CYP1B1. The nature of this CYP1B1 inhibition by PFTalpha was mixed-type with an apparent K(i) of 4.38 nM. Blockage of CYP1 activity by PFTalpha was associated with a decreased metabolism of BP, a reduced formation of BP-derived adducts and a diminished BP-induced apoptosis in human cultured cells targets for PAHs like primary human macrophages and p53-negative KG1a leukaemia cells. These data further substantiate an unexpected and p53 independent action of PFTalpha for preventing toxicity of chemical carcinogens such as PAHs, through inhibition of CYP1 enzyme activities, especially that of CYP1B1. PMID- 16258176 TI - An in vivo analysis of MMC-induced DNA damage and its repair. AB - Mitomycin C (MMC) induces various types of DNA damages that cause significant cytotoxicity to cells. Accordingly, repair of MMC-induced damages involves multiple repair pathways such as nucleotide excision repair, homologous recombination repair and translesion bypass repair pathways. Nonetheless, repair of the MMC-induced DNA damages in mammals have not been fully delineated. In this study, we investigated potential roles for Xeroderma pigmentosum (XP) proteins in the repair of MMC-induced DNA damages using an assay that detects the ssDNA patches generated following treatment with MMC or 8'-methoxy-psoralen (8-MOP) + UVA (ultraviolet light A). Human wild-type cells formed distinctive ssDNA foci following treatment with MMC or 8-MOP + UVA, but not with those inducing alkylation damage, oxidative damage or strand-break damage, suggesting that the foci represent ssDNA patches formed during the crosslink repair. In contrast to wild-type cells, mutant defective in XPE orXPG did not form the ssDNA foci following MMC treatment, while XPF mutant cells showed a significantly delayed response in forming the foci. A positive role for XPG in the repair of MMC induced DNA damages was further supported by observations that cells treated with MMC induced a tight association of XPG with chromatin, and a targeted inhibition of XPG abolished MMC-induced ssDNA foci formation, rendering cells hypersensitive to MMC. Together, our results suggest that XPG along with XPE and XPF play unique role(s) in the repair of MMC-induced DNA damages. PMID- 16258177 TI - Polymorphisms in nucleotide excision repair genes and risk of multiple primary melanoma: the Genes Environment and Melanoma Study. AB - Polymorphisms in six genes involved in nucleotide excision repair of DNA were examined in a large population-based case-control study of melanoma. Genotyping was conducted for 2485 patients with a single primary melanoma (controls) and 1238 patients with second or higher order primary melanomas (cases). Patients were ascertained from nine geographic regions in Australia, Canada, Italy and the United States. Positive associations were observed for XPD 312 Asn/Asn versus Asp/Asp [odds ratio (OR) = 1.5, 95% confidence interval (CI) 1.2-1.9] and XPD 751 Gln/Gln versus Lys/Lys (OR = 1.4, 95% CI 1.1-1.7) genotypes and melanoma. The combined XPD Asn (A) 312 + Gln (C) 751 haplotype was significantly more frequent in cases (32%) compared with controls (29%) (P = 0.003) and risk of melanoma increased significantly with one and two copies of the haplotype (ORs 1.2, 95% CI 1.0-1.4, and 1.6, 95% CI 1.2-2.0, trend P = 0.002). No significant associations were observed for HR23B codon 249, XPG codon 1104, XPC codon 939, XPF codon 415, XPF nt 2063, ERCC6 codon 1213 or ERCC6 codon 1230. ORs for XPD and XPC genotypes were stronger for melanoma diagnosed at an early age, but tests for interaction were not statistically significant. The results provide further evidence for a role of XPD in the etiology of melanoma. PMID- 16258178 TI - The effect of sweetness on the efficacy of carbohydrate supplementation during exercise in the heat. AB - The aim of the present study was to investigate potential mechanisms responsible for the improvement in prolonged exercise capacity in hot environments with exogenous carbohydrate. Eight endurance-trained men (VO(2)max 60.5 +/- 2.4 ml.kg( 1).min(-1), mean +/- SE) cycled to exhaustion on three occasions at 60% VO(2)max at an ambient temperature of 35 degrees C. They ingested either a sweet 6.4% carbohydrate solution (SC), a nonsweet 6.4% carbohydrate solution (NSC), or water (W). Exercise capacity was significantly increased with SC and NSC compared to W, the improvements corresponding to 15.8% and 11.8%, respectively. No difference in exercise capacity was seen between SC and NSC solutions. Plasma glucose concentrations were higher during the SC and NSC trials compared to W, significantly so at 10 min and at fatigue. Rates of carbohydrate oxidation were higher in the SC and NSC trials, although the rates never declined below 2.1 +/- 0.2 g.min(-1) in the W trial. There was no difference in the rate of rise of rectal temperature between trials, but there was a trend for subjects to fatigue at higher temperatures during the two carbohydrate trials. In conclusion, exogenous carbohydrate, independent of sweetness, improves exercise capacity in the heat compared to water alone. PMID- 16258179 TI - Does prior 1500-m swimming affect cycling energy expenditure in well-trained triathletes? AB - The purpose of this study was to determine the effects of a 1,500-m swim on energy expenditure during a subsequent cycle task. Eight well-trained male triathletes (age 26.0 +/- 5.0 yrs; height 179.6 +/- 4.5 cm; mass 71.3 +/- 5.8 kg; VO(2)max 71.9 +/- 7.8 ml.kg(-1).min(-1)) underwent two testing sessions in counterbalanced order. The sessions consisted of a 30-min ride on the cycle ergometer at 75% of maximal aerobic power (MAP), and at a pedaling frequency of 95 rev.min(-1), preceded either by a 1,500-m swim at 1.20 m.s(-1) (SC trial) or by a cycling warm-up at 30% of MAP (C trial). Respiratory and metabolic data were collected between the 3rd and the 5th min, and between the 28th and 30th min of cycling. The main results indicated a significantly lower gross efficiency (13.0%) and significantly higher blood lactate concentration (56.4%), VO(2) (5.0%), HR (9.3%), VE (15.7%), and RF (19.9%) in the SC compared to the C trial after 5 min, p < 0.05. After 30 min, only VE (7.9%) and blood lactate concentration (43.9%) were significantly higher in the SC compared to the C trial, p < 0.05. These results confirm the increase in energy cost previously observed during sprint-distance triathlons and point to the importance of the relative intensity of swimming on energy demand during subsequent cycling. PMID- 16258180 TI - [Comparison of male and female thermal, cardiac, and muscular responses induced by a prolonged run undertaken in a hot environment]. AB - The aim of this study was to compare male and female thermal, cardiac, and muscular responses induced by a prolonged run undertaken in a hot environment. Twelve volunteers participated in this study. The first group consisted of 6 men and the second one consisted of 6 women. After determination of their VO(2)max and maximal aerobic velocity (MAV), each athlete completed a 40-min run at 65% MAV in a hot and dry environment (temperature 31-33 degrees C, relative humidity 30%). Immediately before and after the run, each subject performed two different vertical jumps, i.e., a squat jump (SJ) and a counter-movement jump (CMJ) on a force platform. Force, velocity, power, and jump height were measured during each jump. The completion of the run was associated with a significant loss (p < 0.001) of body mass (BM) and significant increases (p < 0.001) in heart rate, tympanic temperature, and lactate concentration ([La]). Muscle power was significantly improved (+9%, p < 0.05) during the SJ only in the women. A significant enhancement of this parameter was also demonstrated during the CMJ in both groups (men: +10%, p < 0.05; women: +8%, p < 0.01). Surprisingly, a comparison of thermal, cardiac, and muscular responses did not reveal any significant differences between the sexes. Moderate dehydration (-2.0 to -2.3% of BM) and a rise in core temperature (above 39.2 degrees C) induced by the 40-min run led to an improvement of muscular strength in both men and women. However, the results of this study did not reveal any significant between-sex differences in thermal, cardiac, and muscular responses after exercising in the heat. PMID- 16258181 TI - Chronic but not acute oral L-arginine supplementation delays the ventilatory threshold during exercise in heart failure patients. AB - The purpose of this study was to determine, in heart failure patients (HF), whether acute or chronic L-arginine supplementation (LAS) might delay the ventilatory threshold (VT) and whether chronic LAS might reduce exercise-induced plasma lactate increase. HF patients undertook 4 cardiopulmonary bicycle exercises tests. The first 3 were maximal without (EX(1)), after acute (EX(2)), or chronic (EX(3)) oral LAS (6 gm twice a day for 6 weeks). The 4th test (EX(4)) performed after chronic LAS, was similar to the first in order to investigate the effect of chronic LAS on circulating lactate levels. Results showed that acute LAS failed to improve both submaximal and maximal exercise capacities. Similarly, maximal exercise capacity remained unmodified after chronic LAS. Nevertheless, chronic LAS delayed significantly the patients' ventilatory threshold. Thus exercise duration prior to VT increased (mean +/- SEM) from 6.04 +/- 0.9 to 7.7 +/- 1.03 min (p = 0.04), resulting in a significant increase in oxygen uptake (1.05 +/- 0.08 to 1.24 +/- 0.12 L.min(-1); p = 0.03), CO(2) release (0.94 +/- 0.10 to 1.2 +/- 0.12 L.min(-1); p = 0.018), minute ventilation (29.31 +/- 2.8 to 34.5 +/- 2.7 L; p = 0.009), and workload (60.7 +/- 9.8 to 78.5 +/- 10.2 watts; p = 0.034). Furthermore, chronic LAS significantly reduced the exercise-induced increase in postexercise plasma lactate concentration (-21 +/- 7%). In conclusion, unlike acute supplementation, chronic LAS significantly delays the ventilatory threshold, and chronic LAS reduces circulating plasma lactate in HF patients. These data suggest that chronic LAS might improve the ability of HF patients to perform their daily-life activities. PMID- 16258182 TI - Torso stabilization reduces the metabolic cost of producing cycling power. AB - Many researchers have used cycling exercise to evaluate muscle metabolism. Inherent in such studies is an assumption that changes in whole-body respiration are due solely to respiration at the working muscle. Some researchers, however, have speculated that the metabolic cost of torso stabilization may contribute to the metabolic cost of cycling. Therefore, our primary purpose was to determine whether a torso stabilization device would reduce the metabolic cost of producing cycling power. Our secondary purpose was to determine the validity of the ergometer used in this study. Nine male cyclists cycled on a Velotron cycle ergometer at mechanical power outputs intended to elicit 50, 75, and 100% of their ventilatory threshold at 40, 60, and 80 rpm, with and without torso stabilization. Power was controlled by the Velotron in iso-power mode and measured with an SRM powermeter. We determined metabolic cost by indirect calorimetery and recorded power output. Torso stabilization significantly reduced metabolic cost of producing submaximal power (1%), and reduction tended to be greatest at the lower pedaling rates where pedaling force was greatest (1.6% at 40 rpm, 1.2% at 60 rpm, 0.2% at 80 rpm). Power, measured with the SRM powermeter, was strongly correlated with that specified to the Velotron ergometer control unit (R(2) > 0.99). We conclude that muscular contractions associated with torso stabilization elicit significant metabolic costs, which tend to be greatest at low pedaling rates. Researchers who intend to make precise inferences regarding metabolism in the working muscles of the legs may wish to provide torso stabilization as a means of reducing variability, particularly when comparing metabolic data across a wide range of pedaling rates. PMID- 16258183 TI - Vascular nitric oxide and oxidative stress: determinants of endothelial adaptations to cardiovascular disease and to physical activity. AB - Cardiovascular disease is the single leading cause of death and morbidity for Canadians. A universal feature of cardiovascular disease is dysfunction of the vascular endothelium, thus disrupting control of vasodilation, tissue perfusion, hemostasis, and thrombosis. Nitric oxide bioavailability, crucial for maintaining vascular endothelial health and function, depends on the processes controlling synthesis and destruction of nitric oxide as well as on the sensitivity of target tissue to nitric oxide. Evidence supports a major contribution by oxidative stress-induced destruction of nitric oxide to the endothelial dysfunction that accompanies a number of cardiovascular disease states including hypertension, diabetes, chronic heart failure, and atherosclerosis. Regular physical activity (exercise training) reduces cardiovascular disease risk. Numerous studies support the hypothesis that exercise training improves vascular endothelial function, especially when it has been impaired by preexisting risk factors. Evidence is emerging to support a role for improved nitric oxide bioavailability with training as a result of enhanced synthesis and reduced oxidative stress-mediated destruction. Molecular targets sensitive to the exercise training effect include the endothelial nitric oxide synthase and the antioxidant enzyme superoxide dismutase. However, many fundamental details of the cellular and molecular mechanisms linking exercise to altered molecular and functional endothelial phenotypes have yet to be discovered. The working hypothesis is that some of the cellular mechanisms contributing to endothelial dysfunction in cardiovascular disease can be targeted and reversed by signals associated with regular increases in physical activity. The capacity for exercise training to regulate vascular endothelial function, nitric oxide bioavailability, and oxidative stress is an example of how lifestyle can complement medicine and pharmacology in the prevention and management of cardiovascular disease. PMID- 16258184 TI - Maximizing acute fat utilization: effects of exercise, food, and individual characteristics. AB - In discussion of the physiological mechanisms that regulate fat metabolism, and with consideration of the metabolic stimuli that modulate substrate metabolism, the issue of how an acute state of negative lipid balance can be maximized is addressed. The regulation of lipolysis by catecholamines and insulin is reviewed, and the mechanisms of fatty acid mobilization and uptake by muscle are also briefly discussed. The implications of substrate availability and the hormonal response during physiological states such as fasting, exercise, and after food intake are also addressed, with particular regard to the influences on fatty acid mobilization and/or oxidation from eliciting these stimuli conjointly. Finally, a brief discussion is given of both the nature of exercise and the exercising individual, and how these factors influence fat metabolism during exercise. It is also a primary thrust of this paper to underline gaps in the existing literature with regard to exercise timing concerning food ingestion for maximizing acute lipid utilization. PMID- 16258185 TI - Application of design-of-experiments procedures to optimize efficiently pretreatment of lipase for use in a nonaqueous reaction. AB - A variety of different pretreatments can improve the performance of enzymes in nonpolar reaction media. These pretreatments have primarily been studied in isolation; however, interactions between some pairs of pretreatments are known to exist. The presence of these interactions complicates the design of an optimum multifactor pretreatment. Modern design-of-experiments techniques allow the simultaneous optimization of two or more variables. To improve the performance of lipase in a model reaction, we used a technique called the method of steepest ascent to optimize three variables simultaneously: pretreatment pH and sodium phosphate concentration, and the concentration of acetic acid (one of the reactants) in the reaction mixture. In only 26 experimental runs, this optimization process determined a combination of variable settings that yielded a reaction product approx 180 times faster than achieved with untreated enzyme. Evidence is presented to demonstrate that locating this optimum with single factor experiments would be inefficient. This article demonstrates the efficiency of the method of steepest ascent particularly for evaluation of enzymatic reaction conditions exhibiting significant interactions. PMID- 16258186 TI - Conversion of municipal solid waste into carboxylic acids by anaerobic countercurrent fermentation: effect of using intermediate lime treatment. AB - Municipal solid waste (MSW) and sewage sludge (SS) were combined and anaerobically converted into carboxylate salts by using a mixed culture of acid forming microorganisms. MSW is an energy source and SS is a source of nutrients. In this study, MSW and SS were combined, so they complemented each other. Four fermentors were arranged in series for a countercurrent fermentation process. In this process, the solids and liquid were transferred in opposite directions, with the addition of fresh biomass to fermentor 1 and fresh liquid media to fermentor 4. An intermediate lime treatment of solids exiting fermentor 3 before entering fermentor 4 was applied to improve the product acid concentration from the untreated MSW/SS fermentations. All fermentations were performed under anaerobic conditions at 40 degrees C. Calcium carbonate was added to neutralize the carboxylic acids and to control the pH. Iodoform was used as a methanogen inhibitor. Carboxylic acid concentration and gas composition were determined by gas chromatography. Substrate conversion was measured by volatile solids loss, and carboxylic acid productivity was calculated as the function of the total carboxylic acids produced, the amount of liquid in all fermentors, and time. The addition of intermediate lime treatment increased product concentration and conversion by approx 30 and 15%, respectively. The highest carboxylic acid concentrations for untreated MSW/SS fermentations with and without intermediate lime treatment were 22.2 and 17.7 g of carboxylic acid/L of liquid, respectively. These results confirm that adding a treatment step between fermentor 3 and fermentor 4 will increase the digestibility and acid productivity of the fermentation. PMID- 16258187 TI - Evaluation of a new system for developing particulate enzymes based on the surface (S)-layer protein (RsaA) of Caulobacter crescentus: fusion with the beta 1,4-glycanase (Cex) from the cellulolytic bacterium Cellulomonas fimi yields a robust, catalytically active product. AB - Immobilized biocatalysts, including particulate enzymes, represent an attractive tool for research and industrial applications because they combine the specificity of native enzymes with the advantage that they can be readily separated from end product and reused. We demonstrated the use of the Caulobacter crescentus surface (S)-layer protein (RsaA) secretion apparatus for the generation of particulate enzymes. Specifically, a candidate protein made previously by fusion of the beta-1,4-glycanase (Cex) from the cellulolytic bacterium Cellulomonas fimi with the C-terminus of RsaA was evaluated. Cex/RsaA cleaved the glycosidic linkage in the artificial substrate p-nitrophenyl-beta-D cellobioside with a KM similar to that of native Cex (1.1 mM for Cex/RsaA vs 0.60 mM for Cex), indicating that the particulate Cex enzyme was able to bind substrate with wild-type affinity. By contrast, the kcat value was significantly reduced (0.08 s-1 for Cex/RsaA vs 15.8 s-1 for Cex), likely owing to the fact that the RsaA C-terminus induces spontaneous unstructured aggregation of the recombinant protein. Here, we demonstrated that not only can an RsaA fusion protein be cheaply produced and purified to a high yield (76 mg/L of dry wt for Cex/RsaA), but it can also be efficiently recycled. The Caulobacter S-layer secretion system therefore offers an attractive new model system for the production of particulate biocatalysts. PMID- 16258188 TI - Effects of aeration on growth and on production of bacteriocins and other metabolites in cultures of eight strains of lactic acid bacteria. AB - In general, it is accepted that the production of bacteriocins in lactic acid bacteria cultures implies moderately to highly restrictive conditions regarding the availability of oxygen. However, the situation appears to be more complex, probably owing to the facultative anaerobic character of these microorganisms. By studying the culture of eight strains of lactic acid bacteria carried out in vessels with different loads of medium within an interval that determines linearly the minimum availability of oxygen, the existence of three types of behavior was highlighted: production increases (1) with the availability of oxygen, (2) with the restriction of this availability, and (3) toward both extremes of these conditions, diminishing in intermediate situations. These behaviors affected not only the production of bacteriocins, but also their metabolic character (in the Luedeking and Piret sense), as well as the production of other characteristic metabolites, such as lactic acid, acetic acid, and ethanol. PMID- 16258189 TI - Comparison of bacterial and phage display peptide libraries in search of target binding motif. AB - Genetic engineering allows modification of bacterial and bacteriophage genes, which code for surface proteins, enabling display of random peptides on the surface of these microbial vectors. Biologic peptide libraries thus formed are used for high-throughput screening of clones bearing peptides with high affinity for target proteins. There are reports of many successful affinity selections performed with phage display libraries and substantially fewer cases describing the use of bacterial display systems. In theory, bacterial display has some advantages over phage display, but the two systems have never been experimentally compared. We tested both techniques in selecting streptavidin-binding peptides from two commercially available libraries. Under similar conditions, selection of phage-displayed peptides to model protein streptavidin proved convincingly better. PMID- 16258190 TI - Protection of mitochondrial integrity from oxidative stress by selenium containing glutathione transferase. AB - The antioxidant activity of a novel artificial glutathione peroxidase-like enzyme, selenium-containing glutathione 5-transferase from Lucilia cuprina (seleno-LuGST1-1), was studied by using a ferrous sulfate/ascorbate-induced mitochondrial damage model system. Swelling of mitochondria, lipid peroxidation, and cytochrome-c oxidase activity were selected to evaluate the preservation of mitochondrial integrity in this system. Seleno-LuGST1-1 could effectively protect the mitochondria against oxidative damage in a dose-dependent manner and exhibited both higher catalytic activity and greater antioxidant ability than the classic mimic, 2-phenyl-1,2-benziososelenazol-3(2H)-one (Ebselen). This novel artificial biocatalyst therefore may have great potential for pharmacologic application in the treatment of reactive oxygen species-related diseases. PMID- 16258191 TI - Inhibitors of mitogen-activated protein kinases downregulate COX-2 expression in human chondrocytes. AB - Inducible prostaglandin synthase (cyclooxygenase-2, COX-2) is expressed in rheumatoid and osteoarthritic cartilage and produces high amounts of proinflammatory prostanoids in the joint. In the present study we investigated the effects of the inhibitors of mitogen-activated protein kinase (MAPK) pathways Erk1/2, p38, and JNK on COX-2 expression and prostaglandin E2 (PGE2) production in human chondrocytes. Proinflammatory cytokine IL-1beta caused a transient activation of Erk1/2, p38, and JNK in immortalized human T/C28a2 chondrocytes and that was followed by enhanced COX-2 expression and PGE2 production. PD98059 (an inhibitor of Erk1/2 pathway) suppressed IL-1-induced COX-2 expression and PGE2 production in a dose-dependent manner, and seemed to have an inhibitory effect on COX-2 activity. SB203580 (an inhibitor of p38 pathway) but not its negative control compound SB202474 inhibited COX-2 protein and mRNA expression and subsequent PGE2 synthesis at micromolar drug concentrations. SP600125 (a recently developed JNK inhibitor) but not its negative control compound N1-methyl-1,9 pyrazolanthrone downregulated COX-2 expression and PGE2 formation in a dose dependent manner. SP600125 did not downregulate IL-1-induced COX-2 mRNA expression when measured 2 h after addition of IL-1beta but suppressed mRNA levels in the later time points suggesting post-transcriptional regulation. Our results suggest that activation of Erk1/2, p38, and JNK pathways belongs to the signaling cascades that mediate the upregulation of COX-2 expression and PGE2 production in human chondrocytes exposed to proinflammatory cytokine IL-1beta. PMID- 16258192 TI - Changes in serum cytokine levels in active tuberculosis with treatment. AB - It has been reported that IFN-gamma, TNF-alpha, and IL-12 stimulate, and that IL 10, TGF-beta, and IL-4 suppress the protective immune response against tuberculosis. We aim to evaluate changes in the serum levels of pro and antiinflammatory cytokines in active pulmonary tuberculosis (APTB) and the possible effects of treatment on these changes. Serum IL-12p40, IL-4, IL-10, TNF alpha, IFN-gamma, and TGF-beta1 levels were determined in 20 APTB cases (group 1) before and 2, 4, and 6 months after therapy. The same parameters were also determined in 9 inactive pulmonary tuberculosis (IPTB) cases (group 2) and 9 healthy controls (HC, group 3). Before treatment, the mean serum IFN-gamma, TNF alpha, and IL-10 levels in group 1 were statistically higher than those in group 2 (P=.001, P=.024, P=.016, resp) or group 3 (P=.003, P=.002, P=.011, resp). The levels in group 1 decreased significantly after treatment (P=.001 for IFN-gamma, P=.004 for TNF-alpha, P=.000 for IL-10). The serum levels of IL-12p40 were significantly higher in group 1 than in group 3 (P=.012) and decreased insignificantly after treatment. There was no difference in serum IL-4 and TGF beta1 levels among the groups (P>.05). Because the serum IL-12p40, IL-10, TNF alpha, and IFN-gamma levels were high in APTB, we believe that these cytokines have important roles in the immune response to Mycobacterium tuberculosis (M tuberculosis). These parameters could be used in follow-up as indicators of the success of APTB therapy. PMID- 16258193 TI - Characterization of acetyl-CoA: lyso-PAF acetyltransferase of human mesangial cells. AB - Platelet activating factor (PAF) is a potent inflammatory mediator produced by various renal cells and it is implicated in renal pathology. The aim of this study is the characterization of remodeling lyso-PAF acetyltransferase, which is activated under inflammatory conditions, in human mesangial cell. Total membranes of mesangial cells were isolated and enzymatic activity and kinetic parameters were determined by trichloroacetic acid precipitation method. The effect of BSA, divalent cations, EDTA, and various chemicals on the activity of lyso-PAF acetyltransferase was also studied. Various detergents were also tested for the solubilization of the enzyme and only glycerol did not affect its activity. Partial purification of solubilized enzyme preparations of human kidney tissue and mesangial cells was performed on anion exchange column chromatography and native-PAGE electrophoresis and two active fractions were detected. PMID- 16258194 TI - Serum levels of TNF-alpha, IFN-gamma, IL-6, IL-8, IL-12, IL-17, and IL-18 in patients with active psoriasis and correlation with disease severity. AB - Recent progress in the understanding of psoriasis has shown that the regulation of local and systemic cytokines plays an important role in its pathogenesis. The most often used psoriasis score is the psoriasis area and severity index (PASI). A simple laboratory test from a blood sample would be an attractive, patient independent, and observer-independent marker of disease severity. To this end, we evaluated the association of serum levels of some proinflammatory cytokines in vivo and their correlation with severity of psoriasis. The serum levels of cytokines levels were determined with the use of the ELISA method. All mean values except IL-17 levels of patients were significantly higher than those of controls. There was a significant correlation between serum levels of IFN-gamma, IL-12, IL-17, and IL-18, and severity of the disease. Psoriasis can be described as a T-cell-mediated disease, with a complex role for a variety of cytokines, which has led to the development of new immunomodulatory therapies. In this study, serum TNF-alpha, IFN-gamma, IL-6, IL-8, IL-12, and IL-18 levels were significantly higher in active psoriatic patients than in controls. Furthermore, high levels of IFN-gamma, IL-12, and IL-18 correlated with the clinical severity and activity of psoriasis, and those measurements of serum levels of these cytokines may be objective parameters for the disease severity. PMID- 16258195 TI - Regulation of S100A8/A9 (calprotectin) binding to tumor cells by zinc ion and its implication for apoptosis-inducing activity. AB - S100A8/A9 (calprotectin), which is released by neutrophils under inflammatory conditions, has the capacity to induce apoptosis in various cells. We previously reported that S100A8/A9 induces apoptosis of EL-4 lymphoma cells via the uptake of extracellular zinc in a manner similar to DTPA, a membrane-impermeable zinc chelator. In this study, S100A8/A9-induced apoptosis was examined in several cell lines that are weakly sensitive to DTPA, suggesting S100A8/A9 is directly responsible for apoptosis in these cells. Since zinc inhibits apoptosis of MM46, one of these cells, the regulation by zinc of the capacity of S100A8/A9 to bind MM46 cells was studied. When MM46 cells were incubated with S100A8/A9 in standard or zinc-depleted medium, the amounts of S100A8/A9 bound to cells was markedly lower at 3 h than at 1 h. In contrast, when MM46 cells were incubated with S100A8/A9 in the presence of high levels of zinc, binding to cells was the same at 1 and 3 h. When the cells were permeabilized with saponin prior to analysis, a larger amount of cell-associated S100A8/A9 was detected at 3 h. The amount was further increased in cells treated with chloroquine, suggesting that S100A8/A9 was internalized and degraded in lysosomes. Although it has been reported that S100A8/A9 binds to heparan sulfate on cell membranes, the amount of S100A8/A9 bound to MM46 cells was not reduced by heparinase treatment, but was reduced by trypsin treatment. These results suggest that S100A8/A9 induces apoptosis by direct binding to MM46 cells, and that this activity is regulated by zinc. PMID- 16258197 TI - Repeated doses of UVR cause minor alteration in cytokine serum levels in humans. AB - The aim of our study was to compare serum concentration of IL-1beta, IL-6, IL-8, IL-10, and TNF-alpha in 105 healthy volunteers before and after exposure to UVR: 25 subjects (10 days of UVB), 55 (10 days of UVB or solar-simulated radiation, followed by acute UVB dose), and 25 (local high dose of UVB). In all the individuals blood samples were analyzed before and after final irradiation by chemiluminescence assay. After 10 days of UVB irradiation a statistically significant increase in serum concentration only in IL-8 (P<.05) and strong tendency in TNF-alpha (P=.05) were observed. The applied schedules of irradiation have minor impact on serum cytokine level and still a threshold dose of UVR causing systemic immune impairment is unknown. PMID- 16258196 TI - Vascular endothelial growth factor and placenta growth factor in intrauterine growth-restricted fetuses and neonates. AB - The angiogenic factors vascular endothelial growth factor (VEGF) and placenta growth factor (PlGF) are respectively up- and downregulated by hypoxia. We aimed to study circulating levels of the above factors in intrauterine growth restriction (IUGR) and to correlate their levels with the customized centiles of the infants. The study included 25 IUGR and 25 appropriate for gestational age (AGA) full-term, singleton infants and their mothers. Maternal (MS), fetal (UC), and neonatal day 1 (N1) and 4 (N4) blood was examined. MS and N1 PlGF, as well as UC VEGF levels correlated with the customized centiles of the infants (r= 0.39, P=.007, r=0.34, P=.01, and r= -0.41, P=.004, resp). Furthermore, UC, N1, and N4 VEGF levels were higher in girls (r=0.36, P=.01, r=0.33, P=.02, and r=0.41, P=.005 resp). In conclusion, positive and negative correlations of examined factors with the customized centiles of the infant could rely on placental function and intrauterine oxygen concentrations-both being usually lower in IUGR cases-while higher VEGF levels in girls should possibly be attributed to the stimulating action of estrogens. PMID- 16258198 TI - Ischemia-reperfusion injury of adipofascial tissue: an experimental study evaluating early histologic and biochemical alterations in rats. AB - Fat necrosis remains a serious complication in reconstructive flaps. In clinical setting, it is well known that fat tissue is more susceptible to ischemic events. We aimed to evaluate early histological and biochemical changes of adipofascial tissue in an experimental model. An epigastric flap model in rats was used to evaluate the effect of ischemia-reperfusion (I-R) injury on adipofascial tissue. Two groups of animals (one with ischemia alone and other ischemia-reperfusion group) were used to evaluate the degree of histological edema, congestion and extravascular bleeding, and early biochemical alterations within the adipofascial flaps. The biochemical parameters included glutathione (GSH) and malondialdehyde (MDA). In each group, contralateral groin subcutaneous adipose tissue served as control. These evaluations were compared to normal unmanipulated, contralateral abdominal subcutaneous adipose tissue. The ischemia-reperfused flap group showed histologically significantly much edema congestion and bleeding than the control groups (P<.0001). The control group showed less edema in fat tissue than the ischemia-alone group (P<.05). All of the flaps in the ischemia-only group showed significantly less bleeding and edema than I-R group (P<.001). The ratio of MDA/GSH was 33 in control, 37 in ischemia alone, and 82 in ischemia-reperfusion groups, respectively. This study confirms that significant histologic and biochemical alteration occurs after ischemia and ischemia-reperfusion events in adipose tissue. Marked drop in adipose tissue antioxidant levels after I-R suggested that preemptive measures to this decrease should be undertaken in clinical settings. PMID- 16258199 TI - The effects of oxidative stress in urinary tract infection during pregnancy. AB - The purpose of this study was to determine the effect of urinary tract infection (UTI) on antioxidant systems and lipid peroxidation (LPO) levels during pregnancy. We also investigated if these antioxidant systems and LPO levels differed in each trimester. One hundred forty-three nonpregnant women, as a control group, and 77 pregnant women were included in the study. Urine cultures were performed according to standard techniques. Catalase (CAT), superoxide dismutase (SOD), and LPO levels were measured using a spectrophotometer. UTI was observed in 14 of 77 pregnant women and the isolated microorganisms were Escherichia coli, Klebsiella pneumoniae, and Staphylococcus saprophyticus. CAT, SOD, and LPO levels were increased in pregnant women compared with nonpregnant women (P<.01). CAT, SOD activities, and LPO levels were increased from the first trimester to the third trimester in pregnancy without UTI. However, CAT and SOD activities were decreased, LPO levels were increased from the first trimester to the third trimester in pregnancy with UTI (P<.01). Pregnancy causes oxidative stress and also UTI during pregnancy may aggravate oxidative stress. PMID- 16258200 TI - Anaesthesia and the acute phase protein response in children undergoing circumcision. AB - Concentrations of acute phase proteins (CRP: C-reactive protein, albumin) change during surgery. We investigated the acute phase response to circumcision and the effects of anaesthesia on this response. The children were divided into four groups; group 1 (intratracheal general anaesthesia, n=40), group 2 (general anaesthesia with mask, n=20), group 3 (ketamine, n=20), group 4 (local anaesthesia, n=35). Blood samples were obtained, 24 hours before circumcision, after premedication, and 24 hours after circumcision. CRP and albumin before circumcision were comparable for all groups. There was no increase in CRP, and albumin remained steady throughout the study. No difference was observed among the groups, and related to anaesthesia. No responsiveness may be explained with the size of injured tissue or anatomical and histological type of preputium. PMID- 16258201 TI - Fulminant type 1 diabetes-is it an Asian-oriented disease? PMID- 16258202 TI - Light- and heavy-chain deposition disease (LHCDD): difficulty in diagnosis and treatment. PMID- 16258203 TI - Analysis of 645 patients with primary hyperparathyroidism with special references to cholelithiasis. AB - OBJECTIVE The clinical picture of primary hyperparathyroidism (PHPT) has changed during the past 50 years. It is currently unknown whether or not PHPT is associated with an increased risk of cholelithiasis. PATIENTS: To determine the frequency of cholelithiasis in PHPT we analyzed 645 consecutive patients seen at Prague University Hospital from 1992 through 2002 and compared them with a of normocalcaemic control group. METHODS: We investigated 645 patients with proven PHPT (518 female and 127 males aged 20-80 years) during a period of 10 years. To determine the frequency of cholelithiasis in normal population we analyzed 2,015 patients receiving periodic health examination at an outpatient ward from January 1998 to December 1998 (1505 females and 510 males aged 24-85 years). A detailed history, physical examination, biochemical measurements and abdominal ultrasonography were done. RESULTS: Cholelithiasis was proven in 157 of 518 women (30.3%) and in 11 of 127 men (8.66%) with PHPT. Their mean age was 59.67+/-12 years in women and 56.0+/-10 years in men. In the control group 260 of 1505 women (17.27%) and 54 of the 510 men (10.58%) had cholelithiasis. The mean age was 64.55+/-13.8 years in women and 61.2+/-12.4 in men. Only in the case of women, the difference was highly statistically significant (p<0.001). There were no significant differences between the mean values for the serum calcium level, bone alkaline phosphatase, total cholesterol, urinary hydroxyproline and body mass index in hyperparathyroid patients with and without cholelithiasis. However the hyperparathyroid women with cholelithiasis had an increased concentration of parathyroid hormone (236.1+/-56 pg/ml) compared with hyperparathyroid women without cholelithiasis (179.0+/-45 pg/ml), p<0.01. CONCLUSION: The mechanism of PTH associated gallstone formation may involve inhibition of gallblader emptying, hepatic bile secretion and sphincter Oddi motility as well as modification of bile composition. While it might be difficult to prove it seems likely that the association of cholelithiasis with primary hyperparathyroidism in women with a high concentration of parathyroid hormone is more than merely coincidental and from our study it is obvious that a significant association exists. PMID- 16258204 TI - Metabolic syndrome as a predictor of ischemic stroke in elderly persons. AB - OBJECTIVE: To estimate the prevalence and risk of ischemic stroke associated with metabolic syndrome. METHODS AND PATIENTS: In 197 stroke patients (80 cases of atherothrombotic infarction, 97 lacunar infarction, 16 cardioembolic infarction, 4 others) and 356 age- and sex-matched control subjects aged 65 years and older in Seiyo Municipal Nomura Hospital, we investigated the association between metabolic syndrome and risk factor-dependent augmentation of ischemic stroke in subjects with several risk factors for atherosclerosis. The diagnosis of cerebral infarction in each patient was confirmed by CT findings of the brain and metabolic syndrome was defined as at least 3 of the 5 following conditions: visceral obesity, hypertension (HT), hypertriglyceridemia, low HDL-cholesterol and diabetes mellitus (DM). RESULTS: After adjustment for sex, age, and smoking, metabolic syndrome was significantly related to atherothrombotic infarction (odds ratio, 3.08; 95% confidence interval, 1.69-5.61). Of the individual components, visceral obesity, HT and DM were independent risk factors for atherothrombotic infarction. Increased risk for atherothrombotic infarction was also associated with increases in the 5 component conditions of the metabolic syndrome. CONCLUSION: The clustering of cardiovascular risk factors called metabolic syndrome increases the risk of cardiovascular morbidity, and its identification may thus be important in risk assessment and treatment of patients. PMID- 16258205 TI - Serotonin-2A and 2C receptor gene polymorphisms in Japanese patients with obstructive sleep apnea. AB - OBJECTIVE: The serotonin (5-HT) 2A and 2C receptor subtype plays an important role in the maintenance of upper airway stability and normal breathing in obesity. Polymorphisms in the 5-HT 2A receptor gene (HTR2A) and 5-HT 2C receptor gene (HTR2C) are associated with various diseases. The aim of this study was to investigate whether or not the HTR2A/C genotypes are associated with obstructive sleep apnea (OSA). METHODS: The PCR-restriction fragment length polymorphism method was used to determine genotypes of the HTR2A/C gene. The genotype distributions and allele frequencies were statistically analyzed. SUBJECTS: We studied 177 consecutive male patients with excessive daytime somnolence and an apnea plus hypopnea number [apnea-hypopnea index (AHI)] of greater than five per hour of sleep established by full polysomnography. One hundred Japanese men in whom OSA was clinically excluded were randomly selected as a control group. RESULTS: Genotypes and allele frequencies of 102T/C polymorphism of the HTR2A and 796G/C polymorphism of the HTR2C did not differ between controls and patients with OSA. HTR2C polymorphism was considered inappropriate for association studies because of low frequency of the mutant allele. Multiple regression analysis showed that age and body mass index (BMI) were significantly associated with OSA, but HTR2A polymorphisms were not. HTR2A polymorphisms had no significant relationship with AHI or BMI, although further study with more samples will be needed for powerful statistical analyses. CONCLUSIONS: These results indicate that age and BMI, not these polymorphisms, are associated with OSA in this population. PMID- 16258206 TI - Comparison of the clinical courses of the opticospinal and conventional forms of multiple sclerosis in Japan. AB - We evaluated the clinical courses of 216 patients with multiple sclerosis (MS) diagnosed according to the recommended diagnostic criteria of McDonald et al (10). Sixty-five patients clinically displaying selective involvement of the optic nerves and spinal cord were classified as opticospinal MS (OS-MS), while the other 151 showing disseminated involvement of the central nervous system were classified as conventional MS (C-MS). The disease duration did not differ significantly between the two subtypes (11.2 years vs. 11.5 years). In addition to a higher age of onset, female preponderance and higher Kurtzke's expanded disability status scale (EDSS) scores, the OS-MS patients showed a markedly lower frequency of secondary progressive MS than the C-MS patients (4.6% vs. 29.1%, p=0.0001). The EDSS scores of the C-MS patients were significantly correlated with the disease duration, while those of the OS-MS patients were not. Among the C-MS patients, the frequency of secondary progressive MS was significantly more common in patients with a disease duration of more than 10 years than in those with a shorter duration. These results suggest that the irreversible disability in OS-MS is determined by relapses, rather than by chronic progression, whereas C MS has a similar clinical course to MS in Westerners. PMID- 16258207 TI - Low-dose levodopa therapy in Japanese patients with Parkinson's disease: a retrospective study. AB - OBJECTIVE: To investigate the efficacy and the rate of adverse events of chronic low-dose levodopa-carbidopa therapy in Japanese patients with Parkinson's disease (PD). PATIENTS AND METHODS: A total of 92 Japanese PD patients treated with low doses of levodopa from the outset were studied. Both disease-specific motor disabilities and quality of life (QOL) in the patients were evaluated using the Unified Parkinson's Disease Rating Scale (UPDRS) and the Parkinson's Disease 39 Quality of Life Questionnaire (PDQ39), respectively. RESULTS: In the overall patient group, the mean duration of treatment, the mean daily dose of levodopa, the disability scores and the motor scores of UPDRS were 6.2 years, 186.4 mg, 8.0 and 19.2, respectively. The rates of motor fluctuations, dyskinesias and hallucinations were 8.7%, 6.5% and 14.1%, respectively. The mean summary index of PDQ39 scores was 23.1. Patients with motor fluctuations showed a significantly earlier disease onset. Dose of levodopa, age at onset, and treatment duration were not associated with the occurrence of dyskinesias. Patients with hallucination had higher doses of levodopa and dopamine agonist. CONCLUSIONS: Our results demonstrate that chronic administration of a low-dose levodopa preparation can provide satisfactory benefit with a low incidence of motor complications, and can result in good QOL in Japanese patients with PD. The concomitant use of a small amount of dopamine agonist and amantadine from the outset has partly contributed to a reduced dose of levodopa and the lesser occurrence of motor complications. PMID- 16258208 TI - Peritoneal serous papillary adenocarcinoma: report of four cases. AB - We report four cases of peritoneal serous papillary adenocarcinoma (PSPC), a rare disease; all patients had ascites and high levels of serum CA125. Clinical and radiological examinations could not differentiate the disease from peritoneal metastatic tumors and mesothelioma, and histopathological analysis including immunochemistry on the specimen obtained at laparotomy or laparoscopy was necessary for the diagnosis. One patient lived for 58 months with cytoreductive surgery and chemotherapy, and another is still living after 20 months by chemotherapy alone. In patients with peritoneal tumors of unknown origin and a high level of serum CA125, taking PSPC into consideration in the differential diagnosis, histopathological examination should be performed. PMID- 16258209 TI - Development of hepatocellular carcinoma in a woman with HBV- and HCV-negative autoimmune hepatitis with unsatisfactory response to Corticosteroid. AB - A 57-year-old woman was admitted due to a hemorrhage from esophageal varices. Laboratory tests showed liver dysfunction, elevated immunoglobulin G levels and positivity for anti-nuclear antibodies. Serum hepatitis B virus and hepatitis C virus markers were negative. The liver biopsy specimen was compatible with autoimmune hepatitis, and low-dose prednisolone was started. During the follow up, her serum alanine aminotransferase levels continued to fluctuate between 40 and 60 IU/l. Nine years later, hepatocellular carcinoma 20 mm in diameter was detected. This case suggests that hepatocellular carcinoma develops even if serum alanine aminotransferase levels are maintained at less than twice the upper normal limit. PMID- 16258210 TI - Intramural duodenal hematoma after endoscopic therapy for a bleeding duodenal ulcer in a patient with liver cirrhosis. AB - We report a case of intestinal obstruction due to intramural hematoma of the duodenum following therapeutic endoscopy for a bleeding duodenal ulcer in a patient with liver cirrhosis. A 44-year-old man was admitted to our hospital with severe epigastralgia, nausea and tarry stool. Two years previously he had undergone endoscopic sclerotherapy for esophageal varices caused by alcoholic liver cirrhosis. Endoscopy revealed an open ulcer with a bleeding vessel in the duodenal bulb, and sclerotherapy was performed by clipping the vessel and injecting 20 ml of 0.2% epinephrine. His platelet count was 3.5x10(4)/mul. Twelve hours later, he again developed epigastralgia and hypotension. Emergency computed tomography and ultrasonography revealed an intramural hematoma, 15x18 cm in diameter, at the dorsal and lateral duodenum. Endoscopy and upper gastrointestinal series revealed severe stenosis of the duodenal lumen caused by intramural hematoma. He received parenteral feeding for 22 days and within 8 weeks the hematoma was gradually absorbed using conservative management. Intramural duodenal hematoma may be diagnosed as a complication of the endoscopic procedure in a patient with a bleeding tendency, such as liver cirrhosis. PMID- 16258211 TI - Primary hepatic carcinoid tumor with carcinoid syndrome and carcinoid heart disease: a case report of a patient on long-term follow-up. AB - We report a 57-year-old woman with a primary hepatic carcinoid tumor who had been suffering from symptoms due to carcinoid syndrome and carcinoid heart disease for more than 9 years. Long-term injection of octreotide acetate was fairly effective to relieve the symptoms due to carcinoid syndrome, but its tumor contraction effect was not confirmed. The massive ascites in the follow-up period appeared to be diminished to a certain extent by timely chemotherapy including cisplatin. The patient died of multi-organ failure about 9 years after the onset of disease. Autopsy confirmed a primary hepatic carcinoid tumor with limited distant metastases. PMID- 16258212 TI - Severe infundibular pulmonary stenosis and coronary artery stenosis with ventricular tachycardia 24 years after mediastinal irradiation. AB - A 28-year-old man developed severe infundibular pulmonary stenosis (PS), coronary artery stenosis with sustained ventricular tachycardia (VT) 24 years after mediastinal irradiation (total amount of 40 Gray) for non-Hodgkin's lymphoma. Repair of right ventricular outflow tract and coronary artery bypass graft procedure were performed. Infundibular PS was successfully relieved after operation and VT was also controlled by medication. Mediastinal irradiation often causes various cardiac complications after a latent period. Therefore, continuous careful observation is mandatory in patients with the history of mediastinal irradiation. PMID- 16258213 TI - A Chinese patient presenting with clinical signs of fulminant type 1 diabetes mellitus. AB - A 31-year-old Chinese woman suffering from excessive thirst and appetite loss consulted our hospital. Laboratory examination revealed a high blood glucose level and evidence of diabetic ketoacidosis, however, the serum HbA1c was normal. The 24-hour urinary excretion of C-reactive protein and the glucagon stimulation test indicated the loss of endogenous insulin secretion. The patient satisfied the criteria for the diagnosis of fulminant type 1 diabetes mellitus. However, she tested positive for serum anti-IA2 antibody even though the titer was low. We considered it worthwhile reporting this case, because very few cases of fulminant type 1 diabetes mellitus among Chinese people have been published. PMID- 16258214 TI - High dose chemotherapy and stem cell support in a patient of light- and heavy chain deposition disease with abnormal marrow cell surface antigens and no monoclonal protein. AB - A 53-year-old man with nephrotic syndrome and severe renal failure was diagnosed with light- and heavy-chain deposition disease (LHCDD) by renal biopsy. The patient had no monoclonal protein and mild marrow plasmacytosis (6%), but marrow plasma cells expressed CD19(-)CD56+ and predominant monoclonal kappa-chain, indicating plasma cell dyscrasia. Conventional chemotherapy was ineffective and did not improve renal failure. High dose chemotherapy/peripheral blood stem cell transplantation (HDC/PBSCT) was introduced even after hemodialysis to eliminate aberrant clone and normalization of bone marrow cell surface markers. Immunophenotypic analysis of marrow cells facilitates clinical decision making regarding the use of HDC/PBSCT for LHCDD patients without monoclonal protein. PMID- 16258215 TI - Drug-induced isolated visceral angioneurotic edema. AB - A 44-year-old woman on maintenance hemodialysis was admitted to our hospital because of severe abdominal pain. The patient had been medicated with lisinopril and valsartan for hypertension for one month prior to admission. An abdominal computerized scan (CT) showed a dilated and thickened loop of the small bowel with massive ascites and a small nodule in the jejunum. The patient's abdominal pain was thought to be due to isolated visceral angioneurotic edema induced by lisinopril and/or valsartan, and medication of these two drugs was therefore stopped. Her symptoms resolved and an abdominal CT demonstrated almost complete resolution of ascites and of small bowel edema except for a small nodule in the jejunum. A laparoscopic operation was performed to excise the small nodule of the jejunum, and a histological diagnosis of accessory pancreas of the jejunum was made. This is the first report of isolated visceral angioneurotic edema induced by lisinopril and/or valsartan in a patient on maintenance hemodialysis and, moreover, with the association of accessory pancreas of the jejunum. PMID- 16258216 TI - Systemic tumor embolism mimicking gefitinib ('IRESSA')-induced interstitial lung disease in a patient with lung cancer. AB - We describe a 55-year-old man with advanced adenocarcinoma of the lung who received gefitinib ('IRESSA'). After gefitinib administration for 7 months, computed tomography scan of the chest demonstrated diffuse ground glass opacity and he was suspected to have developed gefitinib-induced interstitial lung disease (ILD). However, transbronchial lung biopsy (TBLB) revealed tumor cells in the middle-size lung vessels. Afterwards, multiple infarctions of the brain, spleen and left kidney were detected. Then, he was considered to have developed systemic tumor emboli, a rare complication. The clinical presentation of this patient was difficult to discriminate from that of ILD, and TBLB was useful in the differential diagnosis. PMID- 16258217 TI - Paget-Schroetter syndrome and pulmonary thromboembolism: clinical follow-up over 5 years. AB - A scaffold constructor lost consciousness at work, leading to emergency admission to our hospital. He had noted gradual worsening of exertional dyspnea over the previous 1 month. Chest radiography showed localized oligemia and enlarged hilar vessels. Pulmonary perfusion scintigraphy confirmed the existence of multiple perfusion defects, so a diagnosis of pulmonary thromboembolism was made. Upper and lower limb venography disclosed interruption of the right subclavian vein (so called Paget-Schroetter syndrome). However, there was no difference in appearance between the right and left upper extremities. Five years after starting anticoagulant therapy, his symptoms have resolved, but serial perfusion scintigraphy and upper extremity venography revealed the persistence of abnormalities. In patients with pulmonary thromboembolism, lifestyle factors (especially heavy manual labor) should be considered and the possibility of subclavian vein thrombosis should be kept in mind. PMID- 16258218 TI - Primary alveolar hypoventilation syndrome complicated with antiphospholipid syndrome. AB - A 32-year-old woman was transported to our hospital by ambulance because of loss of consciousness and breathing induced by drug intoxication. After general status was recovered, her arterial blood gas analysis under breathing room air revealed hypercapnia and hypoxemia which were caused by hypoventilation. After exclusion of apparent pulmonary, neuromuscular and central nerve diseases, she was diagnosed with primary alveolar hypoventilation syndrome. She had the complication of antiphospholipid syndrome (APS), suggesting the possibility of small lesions of the brainstem due to APS, which were too small to be detected on CT or MRI; these small lesions could cause injuries to the respiratory center. PMID- 16258219 TI - A Japanese family with ferroportin disease caused by a novel mutation of SLC40A1 gene: hyperferritinemia associated with a relatively low transferrin saturation of iron. AB - Ferroportin disease, autosomal-dominant reticuloendothelial iron overload, may be more prevalent than hemochromatosis in Japan. Hyperferritinemia of 822 ng/ml with 24.8% transferrin saturation of iron was incidentally noted in a 43-year-old man. His iron overload was selective in Kupffer cells of the liver. Subsequently, his father was found to have asymptomatic hyperferritinemia of 2,283 ng/ml with 62.1% saturation. These affected subjects were heterozygous for 1467A>C (R489S) in SLC40A1, and without other mutations of the hemochromatosis genes. Here, we report a Japanese family with ferroportin disease, characterized by hyperferritinemia with relatively low transferrin saturations of iron. PMID- 16258220 TI - Chronic myelogenous leukemia that occurred two years after the diagnosis of adult Still's disease. AB - A 25-year-old Japanese man was diagnosed with steroid-resistant Adult Still's Disease (ASD) in August 2000. No evidence of chronic myelogenous leukemia (CML) had been found during admissions in 2000 and 2001. In August 2002, he was diagnosed with CML with a peripheral white blood count of 69,940/microl and positivity for Philadelphia chromosome and BCR/ABL fusion gene on bone marrow aspiration. No case of CML was reported to develop from ASD. Because a diagnosis of ASD is based on the exclusion of other diseases, we must be cognizant of the possibility of the development of concurrent diseases. PMID- 16258221 TI - Successful high dose glucocorticoid treatment for subacute neuromyelitis optica with systemic lupus erythematosus. AB - A 54-year-old Japanese woman with a 6-year history of systemic lupus erythematosus (SLE) was admitted to our hospital suffering from acute blindness in her right eye. Her condition recovered after steroid pulse therapy, however, 18 months later she suffered from nuchal pain for 2 weeks after which right hemiparesis with urinary incontinence developed. A spinal magnetic resonance imaging (MRI) revealed cord swelling from C2 to C7. She was diagnosed with neuromyelitis optica (NMO) and intravenous steroid administrations were immediately commenced. Her condition promptly improved. This case was unique because the steroid treatment was quite effective for this case of myelitis, which had passed the acute phase. We supposed that, because most of the lesion was not necrotic or demyelinated, but rather showed edematous change caused by vasculitis based on autoimmune pathogenesis, the symptoms progressed rather gradually and improved promptly in response to glucocorticoid treatment. PMID- 16258222 TI - Localized lesions on MRI in a case of hypertensive brainstem encephalopathy. AB - We report a case of hypertensive brainstem encephalopathy (HBE) with unusual magnetic resonance imaging (MRI) findings. A 67-year-old woman presented with high blood pressure and stupor as the only symptoms. MRI revealed lesions localized in the area from the upper medulla oblongata to the lower pons with high fluid-attenuated inversion recovery (FLAIR) and T2-weighted signal intensity, but these were not seen in the whole brainstem and there were no accompanying occipital lobe changes. To our knowledge, no similar case has been reported. The lesions and symptoms dramatically improved after normalization of blood pressure. Severe hypertension that exceeded the range of autoregulation may have resulted in segmental vasodilatation and the increased vascular permeability may have lead to vasogenic edema in the localized areas of the brainstem. PMID- 16258223 TI - Herpes simplex encephalitis presenting with bilateral hippocampal lesions on magnetic resonance imaging, simultaneously complicated by small cell lung carcinoma. AB - We report a 75-year-old man who developed herpes simplex encephalitis (HSE), presenting with bilateral hippocampal lesions on magnetic resonance imaging, and this case was simultaneously complicated by small cell lung carcinoma. We identified a new anti-neuronal antibody in the cerebrospinal fluid of this patient. Our findings suggest that HSE and paraneoplastic limbic encephalitis (PLE) can overlap, and we discuss the relationships of HSE, PLE, and related disorders. PMID- 16258224 TI - Serum amyloid A as a potent therapeutic marker in a refractory patient with polymyalgia rheumatica. AB - We report a patient with polymyalgia rheumatica (PMR) who showed a relapse soon after tapering of oral prednisolone. C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR) were quickly normalized after the re-increase in oral prednisolone, and muscle pain and stiffness gradually improved in parallel with a decrease in serum amyloid A (SAA). Flow cytometry simultaneously demonstrated an increase in CD8+CD25+ cells and a decrease in CD4+CD25+ cells and CD4+CD45RA+ cells. When clinical symptoms remain with negative results for CRP and ESR even after the start of corticosteroid treatment, SAA might be a potent therapeutic marker for disease activity in PMR. PMID- 16258225 TI - Asymptomatic fascioliasis. AB - A 72-year-old Japanese man displayed asymptomatic eosinophilia for 4 months. Computed tomography showed multiple space-occupying lesions in the liver. Zoonotic liver flukes were suspected based on occupational exposure to cattle, serological and radiological findings. Immunological examination was helpful in diagnosing the disease and laparoscopy was crucial in confirming Fasciola hepatica and excluding the possibility of malignant hepatic tumors. Human fascioliasis was finally diagnosed and praziquantel administered. Blood eosinophilia resolved within 4 months and liver tumors almost disappeared within 12 months. From our experience, laparoscopy with liver biopsy is very important for diagnosing human fascioliasis, particularly for asymptomatic fascioliasis. PMID- 16258226 TI - A case of serogroup A meningococcal meningitis: a case possibly imported from China. PMID- 16258227 TI - Bilateral adrenal hemorrhage due to sepsis resulting in acute adrenal crisis. PMID- 16258228 TI - Circulating Reed-Sternberg cells. PMID- 16258229 TI - A newly derived protein from Bacillus subtilis natto with both antithrombotic and fibrinolytic effects. AB - Natto, steamed soybeans fermented by Bacillus subtilis natto, is a traditional Japanese food. We derived a purified protein layer, called NKCP as a trade mark, from B. subtilis natto fermentation. In the present study, we examined the fibrinolytic and antithrombotic effects of NKCP and identified its active component to clarify the fibrinolytic effect of NKCP observed in preliminary clinical trials previously. The active component of NKCP was identified as a 34 kilodalton protein designated bacillopeptidase F. NKCP showed direct degradation of artificial blood clot in saline. The protease activity, accounting for the fibrinolytic effect of NKCP, was examined with a chromogenic substrate for plasmin. Dose-dependent prolongations of both prothrombin time and active partial thromboplastin time were observed in rats with intra-duodenum administration of NKCP. Our in vitro and in vivo studies suggest that NKCP has both a fibrinolytic effect and an antithrombotic effect similar to heparin. Because NKCP is derived from food and has safety data demonstrated by previous animal experiments and preliminary clinical trials, NKCP is considered as safe for clinical use. PMID- 16258230 TI - Identification and expression of frizzled-3 protein in rat frontal cortex after antidepressant and electroconvulsive treatment. AB - The biological basis for the therapeutic mechanisms of depression are still unknown. While performing EST (expressed sequence tag) analysis to identify some molecular machinery responsible for the antidepressant effect, we determined the full-length nucleotide sequence of rat frizzled-3 protein (Frz3) cDNA. Interestingly, Northern blot analysis demonstrated that elevated levels of Frz3 were expressed continually from embryonic day 20.5 to postnatal 4 weeks in developing rat brain. In adult rat brain, Frz3 mRNA was expressed predominantly in the cerebral cortex and hypothalamus and moderately in the hippocampus. Using real-time quantitative PCR, we demonstrated that chronic treatment with two different classes of antidepressants, imipramine and sertraline, reduced Frz3 mRNA expression significantly in rat frontal cortex. Electroconvulsive treatment (ECT) also reduced Frz3 expression. In contrast, antidepressants and ECT failed to reduce Frz2 expression. Additionally, chronic treatment with the antipsychotic drug haloperidol did not affect Frz3 expression. Recently, the Frz/Wingless protein pathway has been proposed to direct a complex behavioral phenomenon. In conclusion, the Frz3-mediated signaling cascade may be a component of the molecular machinery targeted by therapeutics commonly used to treat depression. PMID- 16258231 TI - Tachykinin agonists modulate cholinergic neurotransmission at guinea-pig intracardiac Ganglia. AB - Effects of substance P (SP) and selective tachykinin agonists on neurotransmission at guinea-pig intracardiac ganglia were studied in vitro. Voltage responses of neurons to superfused tachykinins and nerve stimulation were measured using intracellular microelectrodes. Predominant effects of SP (1 microM) were to cause slow depolarization and enable synaptic transmission at low intensities of nerve stimulation. Augmented response to nerve stimulation occurred with 29 of 40 intracardiac neurons (approx. 73%). SP inhibited synaptic transmission at 23% of intracardiac neurons but also caused slow depolarization. Activation of NK(3) receptors with 100 nM [MePhe(7)]neurokinin B caused slow depolarization, enhanced the response of many intracardiac neurons to low intensity nerve stimulation or local application of acetylcholine, and triggered action potentials independent of other stimuli in 6 of 42 neurons. The NK(1) agonist [Sar(9),Met(O(2))(11)]SP had similar actions but was less effective and did not trigger action potentials independently. Neither selective agonist inhibited cholinergic neurotransmission. We conclude that SP can function as a positive or negative neuromodulator at intracardiac ganglion cells, which could be either efferent neurons or interneurons. Potentiation occurs primarily through NK(3) receptors and may enable neuronal responses with less preganglionic nerve activity. Inhibition of neurotransmission by SP is most likely explained by the known blocking action of this peptide at ganglionic nicotine receptors. PMID- 16258232 TI - Role of 20-hydroxyeicosatetraenoic acid (20-HETE) in vascular system. AB - Cytochrome P450s (P450) metabolize arachidonic acid (AA) to hydroxyeicosatetraenoic acids (HETEs) and epoxyeicosatrienoic acids (EETs). Among these eicosanoids, 20-HETE is formed in a tissue and cell-specific fashion and plays an important role in the regulation of vascular tone in the brain, kidney, heart and splanchnic beds. 20-HETE is a potent vasoconstrictor produced in vascular smooth muscle (VSM) cells. It depolarizes VSM by blocking the open-state probability of Ca2+-activated K+-channels. Inhibitors of the formation of 20-HETE block the myogenic response of renal and cerebral arterioles in vitro and autoregulation of renal and cerebral blood flow in vivo. The formation of 20-HETE in vascular smooth muscle is stimulated by angiotensin II, endothelin and norepinephrine and is inhibited by nitric oxide (NO). 20-HETE also stimulates mitogenic and angiogenic responses in vitro and in vivo. Changes in the production of 20-HETE have been observed in ischemic cerebrovascular diseases, cardiac ischemia-reperfusion injury, kidney diseases, hypertension, diabetes, uremia, toxemia of pregnancy. The physiological and pathophysiological role of 20 HETE in the regulation of vascular tone are being revealed by the use of newly developed inhibitors of the synthesis of 20-HETE and 20-HETE analogs. The present review summarizes recent findings implicating a critical role for 20-HETE in altering cardiovascular function in a variety of pathological conditions. PMID- 16258233 TI - Evidence for the involvement of the cyclooxygenase-metabolic pathway in diclofenac-induced inhibition of spontaneous contraction of rat portal vein smooth muscle cells. AB - The effects of diclofenac, a cyclooxygenase (COX) inhibitor, were investigated on spontaneous phasic contractions of longitudinal preparations of the rat portal vein. Diclofenac produced a concentration-dependent decrease in the amplitude of these spontaneous phasic contractions. Diclofenac (30 microM) decreased the amplitude of the spontaneous phasic increase in the F340/F380 ratio of Fura PE3, an indicator of intracellular Ca2+ concentration. It also reduced the number of action potentials in each burst discharge without changing the resting membrane potential of longitudinal smooth muscle cells. The extent of the distribution of Lucifer Yellow injected into a smooth muscle cell was decreased in the presence of diclofenac (30 microM). Both AH6809, a prostanoid EP receptor antagonist, and SQ22536, an adenylate cyclase inhibitor, decreased the amplitude of the spontaneous contractions. On the other hand, neither ozagrel, a thromboxane synthase inhibitor, nor SQ29548, a prostanoid TP receptor antagonist, significantly affected spontaneous contractions. These results indicate that diclofenac inhibits the amplitude of spontaneous contractions of the rat portal vein through inhibition of electrical activity, which may be related to an inhibition of the cyclooxygenase pathway. PMID- 16258234 TI - The effects of flufenamic acid on spontaneous activity of smooth muscle tissue isolated from the guinea-pig stomach antrum. AB - The effects of flufenamic acid were investigated on slow waves, follower potentials and pacemaker potentials recorded respectively from circular smooth muscle cells, longitudinal smooth muscle cells and interstitial cells of Cajal distributed in the myenteric layers (ICC-MY) of the guinea-pig stomach antrum. Flufenamic acid (>10(-5) M) inhibited the amplitude and rate of rise of the upstroke phase of the slow waves, with no marked alteration in their frequency of occurrence. The inhibitory actions of flufenamic acid appeared to be mainly on slow potentials recorded from circular smooth muscle cells, but not on follower or pacemaker potentials. After abolishing spontaneous slow potentials with flufenamic acid, depolarizing current stimuli could evoke slow potentials with an amplitude that was much smaller than in the absence of flufenamic acid, with no significant alteration to the input resistance of the membrane. The time elapsed for the generation of the 2nd component of the slow waves or the slow potentials evoked during depolarizing current pulse stimulation was increased by flufenamic acid. The rate of rise of unitary potentials, but not the frequency of occurrence, was inhibited by flufenamic acid. These results indicate that the inhibitory actions of flufenamic acid appear to be mainly on the circular muscle layer including the interstitial cells of Cajal distributed within the muscle bundles (ICC-IM). Nifedipine-sensitive spike potentials were not inhibited by flufenamic acid. It is concluded that the selective inhibition of the 2nd component of slow waves by flufenamic acid may be mainly due to the inhibition of ion channels, possibly Ca2+-sensitive Cl--channels, activated during generation of slow potentials in the ICC-IM distributed in the circular muscle layer. PMID- 16258235 TI - The effects of stress in response to mirror drawing test trials on the electrogastrogram, heart rate and respiratory rate of human subjects. AB - Electrogastrograms (EGGs), heart and respiratory rates were recorded simultaneously from human subjects and analyzed spectrally by the maximal entropy method. The stress of the mirror drawing test (MDT) significantly increased both the heart and respiratory rates with an associated increase in LF/HF ratios. MDT stress, however, increased (excited groups) or decreased (inhibited groups) the power amplitude and the power ratio, (peak power during MDT)/(peak power during rest), of the 1-cpm (0-2.4 cpm), 3-cpm (2.5-4.9), 6-cpm (5.0-7.4), 8-cpm (7.5 9.9) and 10-cpm (10.0-12.9) spectral group components of the EGG recording of the electrical activity of the gastrointestinal tract. The depression scores of the subjects in the MDT stress excited group were higher and their resting frequencies were significantly lower than those of the MDT stress inhibited groups in each of the epigastric, supra-umbilical and infra-umbilical 6-cpm frequency groups but not in the epigastric 1-cpm frequency group. Anxiety scores of the MDT stress excited groups were significantly higher and their power ratios were also significantly higher than those of the inhibited groups in each of the epigastric 10-cpm, the supra-umbilical 6-cpm and the infra-umbilical 1- and 3-cpm frequency groups. Therefore, depression scores appeared to have inhibitory effects on the resting EGG frequencies, while anxiety scores appeared to have facilitatory effects on the EGG power ratio during MDT stress. It is suggested that analysis of EGG under MDT stress would provide a measure of the stress sensitivity of the electrical activity of the human gastrointestinal tract. PMID- 16258236 TI - Ribosomal crystallography: peptide bond formation, chaperone assistance and antibiotics activity. AB - The peptidyl transferase center (PTC) is located in a protein free environment, thus confirming that the ribosome is a ribozyme. This arched void has dimensions suitable for accommodating the 3' ends of the A-and the P-site tRNAs, and is situated within a universal sizable symmetry-related region that connects all ribosomal functional centers involved in amino-acid polymerization. The linkage between the elaborate PTC architecture and the A-site tRNA position revealed that the A- to P-site passage of the tRNA 3' end is performed by a rotatory motion, which leads to stereochemistry suitable for peptide bond formation and for substrate mediated catalysis, thus suggesting that the PTC evolved by gene fusion. Adjacent to the PTC is the entrance of the protein exit tunnel, shown to play active roles in sequence-specific gating of nascent chains and in responding to cellular signals. This tunnel also provides a site that may be exploited for local co-translational folding and seems to assist in nascent chain trafficking into the hydrophobic space formed by the first bacterial chaperone, the trigger factor. Many antibiotics target ribosomes. Although the ribosome is highly conserved, subtle sequence and/or conformational variations enable drug selectivity, thus facilitating clinical usage. Comparisons of high-resolution structures of complexes of antibiotics bound to ribosomes from eubacteria resembling pathogens, to an archaeon that shares properties with eukaryotes and to its mutant that allows antibiotics binding, demonstrated the unambiguous difference between mere binding and therapeutical effectiveness. The observed variability in antibiotics inhibitory modes, accompanied by the elucidation of the structural basis to antibiotics mechanism justifies expectations for structural based improved properties of existing compounds as well as for the development of novel drugs. PMID- 16258237 TI - Antibody engineering for the development of therapeutic antibodies. AB - Therapeutic antibodies represent one of the fastest growing areas of the pharmaceutical industry. There are currently 19 monoclonal antibodies in the market that have been approved by the FDA and over 150 in clinical developments. Driven by innovation and technological developments, therapeutic antibodies are the second largest biopharmaceutical product category after vaccines. Antibodies have been engineered by a variety of methods to suit a particular therapeutic use. This review describes the structural and functional characteristics of antibody and the antibody engineering for the generation and optimization of therapeutic antibodies. PMID- 16258238 TI - Identification of a rice gene (Bph 1) conferring resistance to brown planthopper (Nilaparvata lugens Stal) using STS markers. AB - This study was carried out to identify a high-resolution marker for a gene conferring resistance to brown planthopper (BPH) biotype 1, using japonica type resistant lines. Bulked segregant analyses were conducted using 520 RAPD primers to identify RAPD fragments linked to the BPH resistance gene. Eleven RAPDs were shown to be polymorphic amplicons between resistant and susceptible progeny. One of these primers, OPE 18, which amplified a 923 bp band tightly linked to resistance, was converted into a sequence-tagged-site (STS) marker. The STS marker, BpE18-3, was easily detectable as a dominant band with tight linkage (3.9cM) to Bph1. It promises to be useful as a marker for assisted selection of resistant progeny in backcross breeding programs to introgress the resistance gene into elite japonica cultivars. PMID- 16258239 TI - A combined approach for locating box H/ACA snoRNAs in the human genome. AB - A novel combined method for locating box H/ACA small nucleolar RNAs (snoRNAs) is described, together with a software tool. The method adopts both a probabilistic hidden Markov model (HMM) and a minimum free energy (MFE) rule, and filters possible candidate box H/ACA snoRNAs obtained from genomic DNA sequences. With our novel method 12 known box H/ACA snoRNAs, and one strong candidate were identified in 30 nucleolar protein genomic sequences. PMID- 16258240 TI - The fission yeast gene encoding monothiol glutaredoxin 5 is regulated by nitrosative and osmotic stresses. AB - Glutaredoxin (Grx) is a small, heat-stable redox protein acting as a multi functional glutathione (GSH)-dependent disulfide oxidoreductase. We have cloned the monothiol Grx5 gene from the genomic DNA of the fission yeast Schizosaccharomyces pombe. It has 1,904 bp, with one intron, and encodes a putative protein of 146 amino acids with a molecular mass of 16.5 kDa. Recombinant Grx5 produced functional Grx in S. pombe cells. NO-generating sodium nitroprusside (SNP, 1.0 and 2.0 mM) and potassium chloride (KCl, 0.2 and 0.5 M) increased the synthesis of beta-galactosidase from a Grx5-lacZ fusion gene, and transcription of Grx5 was also enhanced by SNP and KCl. Synthesis of beta galactosidase from the Grx5-lacZ fusion was lower in Pap1-negative TP108-3C cells than in wild type KP1 cells, and when Pap1 was overproduced in KP1 cells, the level of beta-galactosidase increased. We also found that Pap1 is involved in the induction of Grx5 by SNP and KCl. S. pombe Grx5 may play a crucial role in responses to nitrosative and osmotic stresses. PMID- 16258241 TI - Molecular properties of excitation-contraction coupling proteins in infant and adult human heart tissues. AB - Excitation-contraction coupling (ECC) proteins in the human heart were characterized using human atrial tissues from different age groups. The samples were classified into one infant group (Group A: 0.2-7 years old) and three adult groups (Group B: 21-30; Group C: 41-49; Group D: 60-66). Whole homogenates (WH) of atrial tissues were assayed for ligand binding, 45Ca2+ uptake and content of ECC proteins by Western blotting. Equilibrium [3H]ryanodine binding to characterize the ryanodine receptor (RyR) of the sarcoplasmic reticulum (SR) showed that the maximal [3H]ryanodine binding (Bmax) to RyR was similar in all the age groups, but the dissociation constant (kd) of ryanodine was higher in the infant group than the adult groups. Oxalate-supported 45Ca2+ uptake into the SR, a function of the SR SERCA2a activity, was lower in the infant group than in the adult groups. Similarly, [3H]PN200-110 binding, an index of dihydropyridine receptor (DHPR) density, was lower in the infant group. Expression of calsequestrin and triadin assessed by Western blotting was similar in the infant and adult groups, but junctin expression was considerably higher in the adult groups. These differences in key ECC proteins could underlie the different Ca2+ handling properties and contractility of infant hearts. PMID- 16258242 TI - Microarray profiling of genes differentially expressed during erythroid differentiation of murine erythroleukemia cells. AB - Murine erythroleukemia (MEL) cells are widely used to study erythroid differentiation thanks to their ability to terminally differentiate in vitro in response to chemical induction. At the molecular level, not much is known of their terminal differentiation apart from activation of adult-type globin gene expression. We examined changes in gene expression during the terminal differentiation of these cells using microarray-based technology. We identified 180 genes whose expression changed significantly during differentiation. The microarray data were analyzed by hierarchical and k-means clustering and confirmed by semi-quantitative RT-PCR. We identified several genes including H1f0, Bnip3, Mgl2, ST7L, and Cbll1 that could be useful markers for erythropoiesis. These genetic markers should be a valuable resource both as potential regulators in functional studies of erythroid differentiation, and as straightforward cell type markers. PMID- 16258243 TI - Quercetin inhibits the 5-hydroxytryptamine type 3 receptor-mediated ion current by interacting with pre-transmembrane domain I. AB - The flavonoid, quercetin, is a low molecular weight substance found in apple, tomato and other fruit. Besides its antioxidative effect, quercetin, like other flavonoids, has a wide range of neuropharmacological actions including analgesia, and motility, sleep, anticonvulsant, sedative and anxiolytic effects. In the present study, we investigated its effect on mouse 5-hydroxytryptamine type 3 (5 HT3A) receptor channel activity, which is involved in pain transmission, analgesia, vomiting, and mood disorders. The 5-HT3A receptor was expressed in Xenopus oocytes, and the current was measured with the two-electrode voltage clamp technique. In oocytes injected with 5-HT3A receptor cRNA, quercetin inhibited the 5-HT-induced inward peak current (I(5-HT)) with an IC50 of 64.7 +/- 2.2 microM. Inhibition was competitive and voltage-independent. Point mutations of pre-transmembrane domain 1 (pre-TM1) such as R222T and R222A, but not R222D, R222E and R222K, abolished inhibition, indicating that quercetin interacts with the pre-TM1 of the 5-HT3A receptor. PMID- 16258244 TI - Characterization and regulation of the gene encoding monothiol glutaredoxin 3 in the fission yeast Schizosaccharomyces pombe. AB - Glutaredoxins (Grxs) are thioloxidoreductases which are required for maintaining thiol/disulfide equilibrium in living cells. The Grx3 gene, which encodes one of the three monothiol Grxs in the fission yeast Schizosaccharomyces pombe, was characterized, and its transcriptional regulation studied. Genomic DNA encoding Grx3 was isolated by PCR, and a plasmid pTT3 carrying this DNA was produced. The DNA sequence has 1,267 bp, which would encode a monothiol Grx of 166 amino acids with a molecular mass of 18.3 kDa. The putative protein has 27% homology with Grx5, and contains many hydrophobic amino acid residues in its N-terminal region. S. pombe cells harboring pTT3 had increased Grx activity and enhanced survival on minimal medium plates containing aluminum (5 mM), BSO (0.05 mM), menadione (0.01 mM) or cadmium (0.2 mM). The 568 bp upstream region of Grx3 was fused into the promoterless beta-galactosidase gene of the shuttle vector YEp367R to generate fusion plasmid pMJS10. Potassium chloride (KCl) and metals including aluminum and cadmium enhanced the synthesis of beta-galactosidase from the fusion gene. The synthesis of beta-galactosidase was also enhanced, in a Pap1-dependent manner, by fermentable carbon sources such as glucose (at low concentrations) and sucrose, but not by non-fermentable carbon sources such as ethanol and acetate. Grx3 mRNA increased in response to treatment with BSO. These observations indicate that S. pombe Grx3 is involved in the response to stress, and is regulated by stress. PMID- 16258245 TI - HtrA2 interacts with A beta peptide but does not directly alter its production or degradation. AB - HtrA2/Omi is a mammalian mitochondrial serine protease homologous to the E. coli HtrA/DegP gene products. Recently, HtrA2/Omi was found to have a dual role in mammalian cells, acting as an apoptosis-inducing protein and being involved in maintenance of mitochondrial homeostasis. By screening a human brain cDNA library with A beta peptide as bait in a yeast two-hybrid system, we identified HtrA2/Omi as a binding partner of A beta peptide. The interaction between A beta peptide and HtrA2/Omi was confirmed by an immunoblot binding assay. The possible involvement of HtrA2/Omi in A beta peptide metabolism was investigated. In vitro peptide cleavage assays showed that HtrA2/Omi did not directly promote the production of A beta peptide at the beta/gamma-secretase level, or the degradation of A beta peptide. However, overexpression of HtrA2/Omi in K269 cells decreased the production of A beta40 and A beta42 by up to 30%. These results rule out the involvement of HtrA2/Omi in the etiology of Alzheimer's disease. However, the fact that overexpression of HtrA2/Omi reduces the generation of A beta40 and A beta42 suggests that it may play some positive role in mammalian cells. PMID- 16258246 TI - The ribostamycin biosynthetic gene cluster in Streptomyces ribosidificus: comparison with butirosin biosynthesis. AB - A cluster of genes for ribostamycin (Rbm) biosynthesis was isolated from Streptomyces ribosidificus ATCC 21294. Sequencing of 31.892 kb of the genomic DNA of S. ribosidificus revealed 26 open reading frames (ORFs) encoding putative Rbm biosynthetic genes as well as resistance and other genes. One of ten putative Rbm biosynthetic genes, rbmA, was expressed in S. lividans TK24, and shown to encode 2-deoxy-scyllo-inosose (DOI) synthase. Acetylation of various aminoglycoside aminocyclitol (AmAcs) by RbmI confirmed it to be an aminoglycoside 3-N acetyltransferase. Comparison of the genetic control of ribostamycin and butirosin biosynthesis pointed to a common biosynthetic route for these compounds, despite the considerable differences between them in genetic organization. PMID- 16258247 TI - Expression profiles of apoptosis genes in mammary epithelial cells. AB - To investigate apoptosis in HC11 mammary epithelial cells, we compared the gene expression profiles of actively growing and serum-starved apoptotic cells using a mouse apoptosis gene array and 33P-labeled cDNA prepared from the RNA of the two cultures. Analysis of the arrays showed that expression of several genes such as clusterin, secreted frizzled related protein mRNA (sFRP-1), CREB-binding protein (CBP), and others was higher in the apoptotic cells whereas expression of certain genes including survivin, cell division cycle 2 homolog A (CDC2), and cyclin A was lower. These expression patterns were confirmed by RT-PCR and/or Northern analyses. We compared the expression of some of these genes in the mouse mammary gland under various physiological conditions. The expression levels of genes (clusterin, CBP, and M6P-R) up-regulated in apoptotic conditions were higher at involution than during lactation. On the other hand, genes (Pin, CDC2) downregulated in apoptotic conditions were relatively highly expressed in virgin and pregnant mice. We conclude that certain genes such as clusterin, sFRP-1, GAS1 and CBP are induced in apoptotic mammary epithelial cells, and others are repressed. Moreover, the apoptosis array is an efficient technique for comparing gene expression profiles in different states of the same cell type. PMID- 16258248 TI - Inhibition of NF-IL6 activity by manassantin B, a dilignan isolated from Saururus chinensis, in phorbol myristate acetate-stimulated U937 promonocytic cells. AB - Mannasantin B, a dilignan structurally related to manssantin A, is an inhibitor of NF-kappaB transactivation. In the present study, we found that it inhibited PMA-induced expression of IL-1beta, IL-1beta mRNA, and IL-1beta promoter activity in U937 cells with IC50 values of about 50 nM. It also inhibited NF-IL6- and NF kappaB-induced activation of IL-1beta, with IC50 values of 78 nM and 1.6 microM, respectively, revealing a potent inhibitory effect on NF-IL6. Electrophoretic mobility shift assays showed that manassantin B had an inhibitory effect on DNA binding by NF-IL6, but not by NF-kappaB. Further analysis revealed that transactivation by NF-IL6 was also inhibited. Our results indicate that manassantin B suppresses expression of IL-1beta in promonocytic cells by inhibiting not only NF-kappaB but also NF-IL6 activity. Furthermore, our observations suggest that manassantin B may be clinically useful as a potent inhibitor of NF-IL6 activity. PMID- 16258249 TI - Silencing of NbNAP1 encoding a plastidic SufB-like protein affects chloroplast development in Nicotiana benthamiana. AB - It was previously shown that AtNAP1 is a plastidic SufB protein involved in Fe-S cluster assembly in Arabidopsis. In this study, we investigated the effects of depleting SufB protein from plant cells using virus-induced gene silencing (VIGS). VIGS of NbNAP1 encoding a Nicotiana benthamiana homolog of AtNAP1 resulted in a leaf yellowing phenotype. NbNAP1 was expressed ubiquitously in plant tissues with the highest level in roots. A GFP fusion protein of the N terminal region (M1-V103) of NbNAP1 was targeted to chloroplasts. Depletion of NbNAP1 resulted in reduced numbers of chloroplasts of reduced size. Mitochondria also seemed to be affected. Despite the reduced number and size of the chloroplasts in the NbNAP1 VIGS lines, the expression of many nuclear genes encoding chloroplast-targeted proteins and chlorophyll biosynthesis genes remained unchanged. PMID- 16258250 TI - Biochemical properties and localization of the beta-expansin OsEXPB3 in rice (Oryza sativa L.). AB - Alpha-expansins are bound to the cell wall of plants and can be solubilized with an extraction buffer containing 1 M NaCl. Localization of alpha-expansins in the cell wall was confirmed by immunogold labeling and electron microscopy. The subcellular localization of vegetative beta-expansins has not yet been studied. Using antibodies specific for OsEXPB3, a vegetative beta-expansin of rice (Oryza sativa L.), we found that OsEXPB3 is tightly bound to the cell wall and, unlike alpha-expansins, cannot be solubilized with extraction buffer containing 1 M NaCl. OsEXPB3 protein could only be extracted with buffer containing SDS. The subcellular localization of the OsEXPB3 protein was confirmed by immunogold labeling and electron microscopy. Gold particles were mainly distributed over the primary cell walls. Immunohistochemistry showed that OsEXPB3 is present in all regions of the coleoptile and root tissues tested. PMID- 16258251 TI - Characterization of putrescine uptake in hamster amelanocytic melanoma AMEL-3 cells. AB - The uptake of putrescine, spermidine and spermine by Fortner's hamster amelanocytic melanoma AMEL-3 cells was observed in this study to be time dependent, temperature-sensitive, pH-dependent and saturable. Metabolic poisons nullified polyamine uptake, an indication that this is an energy-requiring mechanism. The presence of Na+ ions was found to be requisite to full activity. Valinomycin, gramicidin, monensin and the calcium ionophore calcimycin were also observed to inhibit the process substantially. The transporter active site would seem to contain sulfhydryl groups. Other diamines and polyamine analogues, as well as cationic diamidines, suppressed putrescine uptake. The presence of the ornithine decarboxylase inhibitor DFMO in the culture medium induced putrescine inflows. Putrescine, in turn, induced the negative expression of the carrier, thus suggesting that this influx mechanism is governed by up/down regulation. The cationic diamidine CGP 40215A and its analogue CGP039937A competitively inhibited putrescine transport, with Ki values of 1.9 and 15 microM, respectively. The role of polyamine uptake in these cultures is discussed. PMID- 16258252 TI - Effects of abiotic stresses on cell cycle progression in tobacco BY-2 cells. AB - Mild stresses such as high temperature (30 degrees C) or a low H2O2 concentration induced transient cell cycle arrest at G1/S or G2/M depending on the cell cycle stage at which the stress was applied. When stresses were introduced during G0 or G1, the G1/S checkpoint was mainly used; when stresses were introduced after S phase, G2/M was the primary checkpoint. The slowing of cell cycle progression was associated with transient delays in expression of A-, B-, and D-type cyclins. The delay in expression of NtcycA13, one of the A-type cyclins, was most pronounced. The levels of expression of Ntcyc29 (a cyclin B gene) and of CycD3-1 differed most depending on the applied stress, suggesting that different cellular adjustments to mild heat and a low concentration of H2O2 are reflected in the expression of these two cyclins. PMID- 16258253 TI - A cyclophilin from Griffithsia japonica has thermoprotective activity and is affected by CsA. AB - Members of the multifunctional Cyp family have been isolated from a wide range of organisms. However, few functional studies have been performed on the role of these proteins as chaperones in red alga. For studying the function of cDNA GjCyp 1 isolated from the red alga (Griffithsia japonica), we expressed and purified a recombinant GjCyp-1 containing a hexahistidine tag at the amino-terminus in Escherichia coli. An expressed fusion protein, H6GjCyp-1 maintained the stability of E. coli proteins up to 50 degrees C. For a functional bioassay for recombinant H6GjCyp-1, the viability of E. coli cells overexpressing H6GjCyp-1 was compared with that of cells not expressing H6GjCyp-1 at 50 degrees C. After high temperature treatment for 1 h, E. coli overexpressing H6GjCyp-1 survived about three times longer than E. coli lacking H6GjCyp-1. Measurement of the light scattering of luciferase (luc) showed that GjCyp-1 prevents the aggregation of luc during mild heat stress and that the thermoprotective activity of GjCyp-1 is blocked by cyclosporin A (CsA), an inhibitor of Cyps. Furthermore, the Cyp-CsA complex inhibited the growth of E. coli under normal conditions. The results of the GjCyp-1 bioassays as well as in vitro studies strongly suggest that Cyp confers thermotolerance to E. coli. PMID- 16258254 TI - Molecular phylogeny and geography of Korean medaka fish (Oryzias latipes). AB - The phylogeny and geography of the medaka (Oryzias latipes) populations of Korea were investigated by analyzing sequence data for the mitochondrial control region. From the 41 haplotypes including 25 Korean haplotypes detected in 64 Korean specimens and data for the Japanese and Chinese populations, phylogenetic and nested clade analyses were executed to examine the phylogeny of haplogroups and the relation of the genetic architecture of the haplotypes to the historical geography of the Korean medaka fish. The analyses suggest that there are two very distinct lineages of Korean medaka, and that these result from reproductive isolation mechanisms due to geographic barriers. The southeastern lineage has experienced recent range expansion to the western region. The northwestern lineage, sister to Chinese populations, showed evidence of internal range expansion with shared haplotypes. PMID- 16258255 TI - Chemical inhibition of Wip1 phosphatase contributes to suppression of tumorigenesis. AB - Wip1 is an amplified oncogene whose deletion causes a tumor resistant phenotype in mice. These observations provide justification for a search for Wip1 chemical inhibitors as potential anticancer drugs. Here we report a group of Wip1 inhibitors with anticancer properties both in vitro and in vivo. In vitro, inactivation of Wip1 reduces the proliferation rate of breast cancer cell lines and enhances growth inhibition caused by doxorubicin. In vivo, administration of Wip1 inhibitors decreases proliferation of xenograph tumors and tumors developed in MMTV-c-Neu transgenic mice. We propose that these agents may serve as lead compounds for the development of anticancer drugs targeting Wip1 phosphatase. PMID- 16258256 TI - Hedgehog signaling pathway and gastric cancer. AB - Hedgehog, WNT, FGF and BMP signaling pathways network together during embryogenesis, tissue regeneration, and carcinogenesis. Aberrant activation of Hedgehog signaling pathway leads to pathological consequences in a variety of human tumors, such as gastric cancer and pancreatic cancer. Endoscopic mucosal resection (EMR), endoscopic submucosal dissection (ESD), surgical gastrectomy and chemotherapy are therapeutic options for gastric cancer; however, prognosis of advanced gastric cancer patient is still poor. Here, Hedgehog signaling pathway in human gastric cancer and its clinical applications will be reviewed. Human SHH, IHH, DHH (Hedgehog homologs), HHAT (Hedgehog acyltransferase), HHIP (Hedgehog-interacting protein), DISP1, DISP2, DISP3 (Dispatched homologs), PTCH1, PTCH2 (Patched homologs), SMO (Smoothened homolog), KIF27, KIF7 (Costal-2 homologs), STK36 (Fused homolog), SUFU (SuFu homolog), DZIP1 (Iguana homolog), GLI1, GLI2 and GLI3 (Cubitus interruptus homologs) are implicated in the Hedgehog signaling. PTCH1, FOXM1 and CCND2 are direct transcriptional targets of Hedgehog signaling. Hedgehog signaling activation leads to cell proliferation through cell cycle regulation. SHH regulates growth and differentiation within gastric mucosa through autocrine loop and FOXL1-mediated epithelial-mesenchymal interaction. SHH is implicated in stem/progenitor cell restitution of damaged gastric mucosa during chronic infection with Helicobacter pylori. SHH up-regulation, IHH upregulation and HHIP down-regulation lead to aberrant activation of Hedgehog signaling through PTCH1 to GLI1 in gastric cancer. Small molecule compounds targeted to SMO (KADD-cyclopamine, SANT1-4, Cur61414) as well as humanized anti SHH antibodies are potent anti-cancer drugs for gastric cancer. Cocktail of Hedgehog inhibitors would be developed as novel therapeutics for gastric cancer. Single nucleotide polymorphism (SNP) and copy number polymorphism (CNP) of Hedgehog signaling genes would be utilized for genetic screening of gastric cancer, while cDNA-PCR, microarray and ELISA detecting aberrant Hedgehog signaling activation would be utilized for therapeutic optional choice. Genetic screening and precise selection of therapeutic options would contribute to the realization of personalized medicine. PMID- 16258257 TI - Effects of polyphyllin D, a steroidal saponin in Paris polyphylla, in growth inhibition of human breast cancer cells and in xenograft. AB - Paris polyphylla is a traditional Chinese Medical herb that has been used in treating cancer for thousands of year. Without studies on the anticancer effects of Paris polyphylla being initiated before, we have first studied the component of Paris polyphylla and have spotted out a steroidal saponin, polyphyllin D. As long as the chemical structure and the improved synthesis of polyphyllin D were ascertained, both in vitro to in vivo studies were performed. It was found that treatment of MCF-7 and MDA-MB-231 cells with polyphyllin D resulted in the inhibition of viability and induction of apoptosis in a dose-dependent manner, with an IC50 of 5 microM and 2.5 microM, respectively, after 48 hours of incubations. Apoptosis of MCF-7 and MDA-MB-231 cells by polyphyllin D was evidenced by the occurrence of DNA fragmentation, formation of a hypodiploid peak in the cell cycle analysis, phosphatidyl-serine externalization and a late loss of membrane integrity. Mechanistically, polyphyllin D dissipates the mitochondrial membrane potential, induces a downregulation of anti-apoptotic Bcl 2 expression and an up-regulation of pro-apoptotic Bax expression, and activates caspase-9. These results suggest that polyphyllin D elicits apoptosis through mitochondria dysfunction. In vivo study demonstrated that daily administration of polyphyllin D (2.73 mg/kg body weight) through intravenous injection for ten days in nude mice bearing MCF-7 cells effectively reduced tumor growth for 50% in terms of tumor weight and size, given no significant toxicity in heart and liver to the host. All these findings provide novel insights that polyphyllin D could serve as a candidate in breast cancer treatment. PMID- 16258258 TI - Avoiding glucocorticoid administration in a neurooncological case. AB - Restricting glucocorticoid (GC) use in the treatment of patients with a solid tumor may help improving outcome. Here, we report administration of celecoxib rather than dexamethasone to prevent brain edema in a patient with a cerebellar glioblastoma multiforme WHO grade IV (GBM) upon the patient's request, as well as determining cerebrospinal fluid (CSF) and serum concentrations. CSF concentration (0.04 microM) was 54 times below serum concentration (2.18 microM), or 2500 times below levels inhibiting GBM cells in vitro (100 microM), revealing a blood CSF barrier for celecoxib. The patient did not require dexamethasone for the entire treatment. GC administration hence was avoided successfully in this case. The role of COX-2 inhibitors in treatment of GBM is detailed, leading to the conclusion of a pressing need for a clinical evaluation of non-steroidal COX-2 inhibitors with the ability to penetrate into brain tumors. PMID- 16258259 TI - Expression of osteopontin and HGF/met in adult soft tissue tumors. AB - There is a need for molecular markers that predict biological behavior of adult soft tissue tumors. Elevated levels of osteopontin (OPN) a transformation-linked protein, have been associated with poor survival in many cancers. OPN induces cell migration in cancer cells, in part through activation of the hepatocyte growth factor (HGF) receptor (Met) and its signaling pathway. Met expression has been associated with a poor prognosis in some sarcomas. In a series of 15 patients with adult soft tissue tumors, we found that mRNA levels of OPN (p=0.015), Met (p=0.03) and HGF (p<0.001) were significantly higher in tumor tissues relative to paired normal tissues. By immunohistochemistry, in tumor tissue from 33 patients, we demonstrated that increased expression of OPN, but not Met protein, was associated with higher stage (p=0.025) and grade (p=0.005). We found that increased expression of OPN, but not Met protein, was associated with decreased overall survival (p=0.008) at five years. This study, which is the first to examine coexpression of these two markers, suggests that OPN may have potential as a prognostic marker in adult soft tissue sarcomas, and further that OPN+/-Met signaling pathways may contribute to their biological behavior. PMID- 16258260 TI - Adenovirus-mediated Ink4a/ARF gene transfer significantly suppressed the growth of pancreatic carcinoma cells. AB - OBJECTIVES: To determine the effects of adenovirus-mediated transfer of p14(ARF) and p16(INK4a) on growth and apoptosis of human pancreatic carcinoma cell lines. RESULTS: Pancreatic carcinoma cell lines, PC-7, PANC-1 and MIA PaCa-2 (p14(ARF)-/ and p16(INK4a)-/-), were used. PC-7 (p53 wt) and MIA PaCa-2 (p53 mt) cells infected with recombinant adenovirus vector expressing p14(ARF) gene (Adp14) showed significant inhibition of cell growth compared with control vector in vitro and in vivo, whereas PANC-1 cells (p53 mt) did not show such effects. Those three cell lines exhibited growth retardation and senescence phenotype after infected with the adenovirus vector containing p16(INK4a) gene (Adp16) in vivo and in vitro. CONCLUSIONS: Adenovirus-mediated transfer of p14(ARF) and p16(INK4a) produces significant growth suppression of pancreatic carcinoma cells in vitro and in vivo. The effects of Adp14 partly depend on the status of p53 gene in those cells. PMID- 16258261 TI - Wnt signaling can repress thrombospondin-1 expression in colonic tumorigenesis. AB - The induction of new blood vessels is critical to the pathogenesis of colon cancer, and inhibition of vascular endothelial growth factor (VEGF) has proven to be an effective approach to the treatment of this malignancy. Another potential therapeutic strategy would utilize endogenous anti-angiogenic molecules such as thrombospondin-1 (TSP-1). However, the regulation of TSP-1 expression in colon cancer is poorly understood. Our results demonstrate that TSP-1 is strongly expressed in normal colonic epithelial cells. However, loss of TSP-1 was observed in early colonic adenomas and it became undetectable in invasive colon cancers. Activation of the Wnt signaling pathway in intestinal epithelial cells repressed TSP-1 gene expression, and inhibition of Wnt signaling in colon cancer cells reversed this repression. Although mutant K-ras also inhibited the TSP-1 promoter in intestinal epithelial cells, silencing of mutant K-ras in colon cancer cells with an activated Wnt pathway did not upregulate TSP-1 expression. Collectively, these findings suggest that activation of the Wnt pathway rather than K-ras plays a more important role in the downregulation of TSP-1 observed in colon cancer. PMID- 16258262 TI - Expression of delayed rectifier potassium channels and their possible roles in proliferation of human gastric cancer cells. AB - Voltage-gated potassium (Kv) channels have been reported to be involved in the proliferation of many types of cells, including tumor cells. The overexpression of the Kv channels and related channel activity are involved in the neoplastic process. Our previous study has shown the existence of delayed rectifier potassium (I(K)) current in gastric cancer cells SGC7901. However, the expression and function of most delayed rectifier potassium (K(D)) channel subunits in gastric cancer cells are not completely resolved. Here we examine expression of K(D) channel subunits in Kv1-Kv3 families in immortalized gastric epithelial cells GES and various gastric cancer cells (including AGS, KATOIII, MKN28, MKN45, MGC803, SGC7901, SGC7901/ADR and SGC7901/VCR), and their roles in cell proliferation. RT-PCR analysis reveals that all cell lines examined express Kv1.3, Kv1.5, Kv1.6, Kv2.1 and Kv2.2. However, Kv1.2 and Kv3.2 genes are barely detectable in any given cancer cell lines. Kv1.5 protein, high mRNA levels in all cell lines examined, is also expressed in some cancer cells lines and more frequently detected in gastric cancer tissues. Downregulation of the expression of Kv1.5 in SGC7901 with RNA interference significantly inhibited the proliferation and tumorigenicity of SGC7901 cells. Moreover, in Ca(2+)-containing rather than Ca(2+)-free medium, KCl (50mM) stimulated a rapid increase in the concentration of cytosolic calcium in empty vector transfected cells that was blocked by verapamil. Likewise, decrease the expression of Kv1.5 with short interfering RNA also blocked the depolarization-induced influx of Ca(2+). This finding suggests that more than one kind of K(D) channel subunits are expressed in various gastric cancer cell lines. Kv1.5 may be involved in tumor cells proliferation by controlling Ca(2+) entry, and the interference of K(D) channels expression and/or activity could provide a novel strategy to reverse the malignant phenotype of gastric cancer cells. PMID- 16258263 TI - Evaluation of irinotecan plus paclitaxel in patients with advanced non-small cell lung cancer. AB - PURPOSE: We conducted a phase II study to evaluate the efficacy and safety of the combination of irinotecan and paclitaxel in patients with advanced stage non small cell lung cancer (NSCLC). PATIENTS AND METHODS: Patients were eligible if they had histologically confirmed chemotherapy naive stage IV NSCLC or stage IIIB disease that was not suitable for combined modality therapy. Patients were treated with irinotecan 50 mg/m2 and paclitaxel 75 mg/m2 on days 1 and 8 of a 21 day cycle. If the patient did not experience >grade 1 toxicity during the first cycle, the dose of irinotecan could be escalated to 60 mg/m(2). Patients were evaluated for tumor response rate, time to progression (TTP), overall survival (OS) and toxicity. RESULTS: Twenty-three eligible patients were treated. Two (9%) patients achieved a partial response. Eight patients (35%) had stable disease. The median number of cycles given per patient was four (range 1-29). The major toxicities were grade >or=3 neutropenia (26%) and grade 3 diarrhea (5%). The median time to progression was 2.8 months (range 0.5-21.8 months) for all patients and 4.3 months for the patients who had either stable disease or a partial response. The median overall survival was 9.2 months (range 0.5-40 months). The one- and two-year survival rates were 39% and 13%, respectively. CONCLUSION: The combination of irinotecan and paclitaxel is safe in advanced NSCLC and affords a survival similar to other non-platinum as well as platinum based doublets. However, this combination does not have sufficient activity to justify further study in an unselected population. If biomarkers are developed that can guide the selection of chemotherapy in an individual patient, there may be a rationale for further evaluation of this regimen. PMID- 16258264 TI - Induction of resistance to Aplidin in a human ovarian cancer cell line related to MDR expression. AB - Aplidin-resistant IGROV-1/APL cells were derived from the human ovarian cancer IGROV-1 cell line by exposing the cells to increasing concentration of Aplidin for eight months, starting from a concentration of 10 nM to a final concentration of 4 microM. IGROV-1/APL cell line possesses five fold relative resistance to Aplidin. IGROV-1/APL resistant cell line shows the typical MDR phenotype: (1) increased expression of membrane-associated P-glycoprotein, (2) cross-resistance to drugs like etoposide, doxorubicin, vinblastine, vincristine, taxol, colchicin and the novel anticancer drug Yondelis (ET-743). The Pgp inhibitor cyclosporin-A restored the sensitivity of IGROV-1/APL cells to Aplidin by increasing the drug intracellular concentration. The resistance to Aplidin was not due to the other proteins, such as LPR-1 and MRP-1, being expressed at the same level in resistant and parental cell line. The finding that cells over-expressing Pgp are resistant to Aplidin was confirmed in CEM/VLB 100 cells, that was found to be 5-fold resistant to Aplidin compared to the CEM parental cell line. PMID- 16258265 TI - Evidence that the yeast spindle assembly checkpoint has a target other than the anaphase promoting complex. AB - The spindle assembly checkpoint monitors biorientation of chromosomes on the metaphase spindle and inhibits the Anaphase Promoting Complex (APC) specificity factor Cdc20. If APC-Cdc20 is the sole target of the spindle checkpoint, then cells lacking APC and its targets, B-type cyclin and securin, would lack spindle checkpoint function. We tested this hypothesis in yeast cells that are APC-null. Surprisingly, we find that such yeast cells are able to activate the spindle assembly checkpoint, delaying cell cycle progression in G2/M phase. These data suggest that the spindle checkpoint has a non-APC target that can restrain anaphase onset. PMID- 16258266 TI - BRCA1 regulates gene expression for orderly mitotic progression. AB - Germline mutations of the BRCA1 gene confer an increased risk for breast cancer and ovarian cancer. To study the contribution of BRCA1 to sporadic cancers, which often exhibit reduced BRCA1 expression, we tested the effect of knocking down BRCA1 on gene expression in human prostate (DU-145) and breast (MCF-7) cancer cells. DNA microarray and confirmatory RNA analyses revealed that BRCA1 small interfering (si) RNA caused down-regulation of multiple genes implicated in the mitotic spindle checkpoint (eg., BUB1B, HEC, and STK6), chromosome segregation (eg., ESPL1, NEK2, and PTTG1), centrosome function (eg., ASPM), cytokinesis (eg., PRC1, PLK, and KNSL2), and the progression into and through mitosis (eg., CDC2, and CDC20). Cells treated with BRCA1-siRNA showed attenuation of the mitotic spindle checkpoint; but not several G2 checkpoints. Finally, BRCA1 knockdown caused the accumulation of multinucleated cells, suggesting a defect in cytokinesis. We conclude that BRCA1 regulates gene expression for orderly mitotic progression. PMID- 16258267 TI - Mitotic entry: a matter of oscillating destruction. AB - Entry into mitosis is essentially driven by cyclin B1 and its associated catalytically active partner Cdk1. While cyclin B1 is kept cytoplasmic throughout interphase, nuclear accumulation occurs just prior to mitosis. This restriction is thought to be part of an oscillating mechanism to properly time mitotic entry. A novel nuclear SCF-type mammalian E3 ligase defined by the F-box containing protein NIPA (nuclear interaction partner of ALK), SCFNIPA, targets nuclear cyclin B1 in interphase while it allows for accumulation at G2/M. Thus, oscillating ubiquitination of nuclear cyclin B1 driven by the SCFNIPA complex contributes to the timing of mitotic entry in the mammalian cell cycle. PMID- 16258268 TI - Regulation of the cyclin-dependent kinase inhibitor p57Kip2 expression by p63. AB - The cyclin-dependent kinase (CDK) inhibitor p57Kip2 is a negative regulator of cell proliferation, binding to a variety of cyclin-CDK complexes and inhibiting their kinase activities. The p57Kip2 gene was recognized as a target gene for p73beta, one member of the p53 family. In spite of this, the phenotypes of p73 and p57Kip2 knockout mice do not resemble each other while there is a phenotypic overlap between the p57Kip2 null mice, the p63 null mice and patients affected by p63 associated syndromes, suggesting that p57Kip2 could be indeed a downstream target of p63. By ChIP we determined that in the HaCaT cell line the DeltaNp63alpha protein is associated to three different regions of the p57Kip2 gene. DeltaNp63 can activate both the endogenous p57Kip2 gene and a reporter vector containing a -2191 promoter fragment of the p57Kip2 gene. Natural p63 mutants, associated to the AEC syndrome, show a partial or complete lack of transactivation potential of the p57Kip2 promoter, while three other natural p63 mutants, associated to the EEC, LMS and SHFM-4 syndromes, were less affected. These data suggests that p63 play an important role in the regulation of p57Kip2 expression and that this regulation is subverted in AEC p63 mutants. PMID- 16258269 TI - Nuclear localized phosphorylated FADD induces cell proliferation and is associated with aggressive lung cancer. AB - Fas-associated death domain (FADD)/Mort1 was initially reported as a pro apoptotic adaptor molecule that recruits the initiator caspases 8 and 10 to promote formation of the death-inducing signal complex (DISC) and mediates receptor induced apoptosis. Recent studies have brought to light ancillary death receptor induced apoptosis-independent activities of FADD that include cell cycle regulation, NF-kappaB activation, cell proliferation and role during embryonic development. We have recently shown that in lung adenocarcinomas increased FADD mRNA and protein are significantly associated with poor survival and that FADD overexpression was not due to gene amplification and/or mutation. In this study we showed that the nuclear localization of FADD and elevated expression of the phosphorylated form of FADD (p-FADD) correlated most closely with an increase in NF-kB activity and poor clinical outcome. These results suggest that levels of p FADD may be used as a prognostic biomarker for predicting survival of lung cancer patients. PMID- 16258270 TI - Teratogens as anti-cancer drugs. AB - Most anticancer drugs are teratogens, merely because they target vital cellular functions. Conversely, some plants produce agents that intentionally target embryonic signaling pathways, precisely to cause birth defects if pregnant animals eat such plants. Cyclopamine, a teratogen produced by a flowering plant, inhibits the Hh/Gli pathway, causing developmental defects such as cyclopia (one eye in the middle of the face). In theory, selective teratogens may suppress cancer cells that reactivate embryonic pathways, while sparing most normal cells. I discuss the potential (and limits) of teratogens in cancer therapy, linking diverse topics from morning sickness of pregnancy, embryonic pathways and poisonous plants to the mechanism of action of anticancer teratogens and their combinations with less selective cytotoxic agents. PMID- 16258271 TI - Cancer pharmacogenomics may require both qualitative and quantitative approaches. AB - Resistance to chemotherapy is a major cause of mortality in patients receiving treatment for most types of cancer, and overcoming drug resistance has become an important focus of current research. A major clinical challenge is the fact that most anticancer drugs have a narrow therapeutic range, that is, their effective dose is relatively close to that associated with substantial toxicity. Significant advances have been achieved in event-free survival of patients with many types of cancer (most dramatically childhood acute lymphoblastic leukemia, ALL) through a better understanding of the pathobiology of human cancers, the cellular mechanisms of cancer chemotherapy, and the determinants of inter individual differences in drug effects and treatment response. It is anticipated that expanding our knowledge of these areas will lead to the development of new anticancer agents and to more effective use of existing cancer chemotherapy. Pharmacogenomics research aims to elucidate the genetics determinants of drug efficacy and toxicity. Results of recent studies indicate that both qualitative and quantitative genomic analyses may be required for precise pharmacogenomic characterization of some types of human cancer. PMID- 16258272 TI - What is the role of tubular epithelial cell apoptosis in polycystic kidney disease (PKD)? AB - Tubular epithelial cell apoptosis occurs in most animal models of polycystic kidney disease (PKD) and in kidneys from humans with autosomal dominant polycystic kidney disease (ADPKD). Induction of apoptosis in cultured tubular epithelial cells results in cyst formation. Induction of apoptosis in the kidney in Bcl-2 deficient mice results in increased proliferation of tubular epithelium and cyst formation. Caspase inhibition reduces tubular apoptosis and proliferation and slows disease progression in the Han:SPRD rat model of PKD. Thus, there is evidence that both epithelial cell apoptosis and proliferation are dysregulated in ADPKD and may represent a general mechanism for cyst growth. PMID- 16258273 TI - Growth control under stress: mTOR regulation through the REDD1-TSC pathway. AB - Dysregulated signaling by the checkpoint kinase TOR (target of rapamycin) has been linked to numerous human cancers. The tuberous sclerosis tumor suppressors TSC1 and TSC2 form a protein complex that integrates and transmits cellular growth factor and stress signals to negatively regulate TOR activity. Several recent reports have identified the stress response gene REDD1 as an essential regulator of TOR activity through the TSC1/2 complex in both Drosophila and mammalian cells. REDD1 is induced in response both to hypoxia and energy stress, and cells that lack REDD1 exhibit highly defective TOR regulation in response to either of these stress signals. While the precise mechanism of REDD1 function remains to be determined, the finding that REDD1-dependent TOR regulation contributes to cell growth/cell size control in flies and mammals suggests that abnormalities of REDD1-mediated signaling might disrupt energy homeostasis and/or promote tumorigenesis. PMID- 16258274 TI - A novel role of p38 alpha MAPK in mitotic progression independent of its kinase activity. AB - Activation of p38alpha MAPK triggers G2/M checkpoint, thus inhibiting cell proliferation. In this study we found that depletion of p38alpha by RNAi also inhibited cell proliferation and caused mitotic arrest. However, treatment with selective small molecule p38 kinase inhibitors had no effect on cell cycle progression, even though the p38 kinase was completely inhibited, revealing p38alpha functions that are independent of its kinase activity. Indeed, ectopic expression of a kinase negative p38alpha rescued the lethality caused by RNAi depletion of the endogenous p38alpha, thus providing further evidence for a kinase-independent function of p38alpha. In addition, we showed that overexpression of the wild type or kinase-negative p38alpha also strongly inhibited cell proliferation, similarly as RNAi depletion of p38alpha. Together the results demonstrate that, in addition to its kinase-dependent functions, such as in activation of G2/M checkpoint, p38alpha also has an essential, kinase independent function. PMID- 16258275 TI - Oncogenic beta-catenin signaling networks in colorectal cancer. AB - Beta-catenin has two distinct functions, namely, maintaining cell-to-cell adhesion and mediating the Wnt/beta-catenin signal transduction pathway, which plays pivotal roles in embryogenesis and in malignant transformation of cells. The oncogenic properties of Wnt/beta-catenin signaling stem from alteration in phosphorylation-dependent protein degradation and subcellular localization of beta-catenin from cell membrane to the nucleus, where it binds to T-cell factor (Tcf) to form a bipartite transcription factor. The beta-catenin/Tcf complex facilitates transcription of target genes that encode effectors for activation of cell proliferation and invasion and inhibition of apoptosis, leading to colorectal cancer development. In addition, in the tumor invasion front, stabilized and activated beta-catenin interacts with other molecular pathways to facilitate tumor progression. This review highlights the beta-catenin-dependent oncogenic signaling network involved in the multi-step process of colorectal tumorigenesis. Wnt signaling evidently regulates stem cells, leading them to differentiate or self-renew. We address roles of oncogenic beta-catenin signaling in the microenvironment of the tumor-host interface that determine the individual tumor's malignant potential and in regulation of putative cancer stem or progenitor cells that represent plausible targets for cancer eradication. PMID- 16258276 TI - Novel functional features of the Lis-H domain: role in protein dimerization, half life and cellular localization. AB - The presence of a conserved protein motif usually implies common functional features. Here, we focused on the LisH (LIS1 homology) domain, which is found in multiple proteins, and have focused on three involved in human genetic diseases; LIS1, Transducin beta-like 1X (TBL1) and Oral-facial-digital type 1 (OFD1). The recently solved structure of the LisH domain in the N-terminal region of LIS1 depicted it as a novel dimerization motif. Our findings indicated that the LisH domain of both LIS1 and TBL1 is essential for in vitro oligomerization. Furthermore, our study disclosed novel in vivo features of the LisH motif. Mutations in conserved LisH amino acids significantly reduced both the protein half-life of LIS1, TBL1, and OFD1, and dramatically affected specific intracellular localizations of these proteins. LIS1 mutated in the LisH domain induced its localization to the actin filaments. TBL1 mutated in the LisH domain was not imported into the nucleus. Mutations in OFD1 modified its localization to the Golgi apparatus and in some cases also to the nucleus. In summary, the LisH domain may participate in protein dimerization, affect protein half-life, and may influence specific cellular localizations. Our results allow the prediction that mutations within the LisH motif are likely to result in pathogenic consequences in genes associated with genetic diseases. PMID- 16258277 TI - Cell cycle sibling rivalry: Cdc2 vs. Cdk2. AB - It has been long believed that the cyclin-dependent kinase 2 (Cdk2) binds to cyclin E or cyclin A and exclusively promotes the G1/S phase transition and that Cdc2/cyclin B complexes play a major role in mitosis. We now provide evidence that Cdc2 binds to cyclin E (in addition to cyclin A and B) and is able to promote the G1/S transition. This new concept indicates that both Cdk2 and/or Cdc2 can drive cells through G1/S phase in parallel. In this review we discuss the classic cell cycle model and how results from knockout mice provide new evidence that refute this model. We focus on the roles of Cdc2 and p27 in regulating the mammalian cell cycle and propose a new model for cell cycle regulation that accommodates these novel findings. PMID- 16258278 TI - ATR activation necessary but not sufficient for p53 induction and apoptosis in hydroxyurea-hypersensitive myeloid leukemia cells. AB - Hydroxyurea (HU) is a competitive inhibitor of ribonucleotide reductase that is used for the treatment of myeloproliferative disorders. HU inhibits DNA replication and induces apoptosis in a cell type-dependent manner, yet the relevant pathways that mediate apoptosis in response to this agent are not well characterized. In this study, we employed the human myeloid leukemia 1 (ML-1) cell line as a model to investigate the mechanisms of HU-induced apoptosis. Exposure of ML-1 cells to HU caused rapid cell death that was accompanied by hallmark features of apoptosis, including membrane blebbing, phosphatidylserine translocation, and caspase activation. HU-induced apoptosis required new protein synthesis, was induced by HU exposures as short as 15 min, and correlated with the accumulation of p53 and induction of the p53 target gene PUMA. p53 induction in ML-1 cells was ATR dependent and downregulation of p53 through RNAi delayed HU induced apoptosis. HU did not induce p53 or induce apoptosis in Molt-3 leukemia cells, even though exposure to HU induced a comparable level of DNA damage and robustly activated the ATR pathway. The microtubule inhibitor nocodazole suppressed HU-induced p53 accumulation in ML-1 cells suggesting that a microtubule-dependent event contributes to p53 induction and apoptosis in this cell line. Our findings outline an HU-induced cell death pathway and suggest that activation of ATR is necessary, but not sufficient, for stabilization of p53 in response to DNA replication stress. PMID- 16258279 TI - Fine-tuning the chromosome ends: the last base of human telomeres. AB - Telomeres protect chromosomes from degradation and loss of vital sequence, block end-end fusion, and allow the cell to distinguish between broken ends and chromosome ends. Mammalian telomeres end in single-stranded (TTAGGG)-rich 3' overhangs that are tucked back into the preceding double stranded region to form a T-loop. The end structure of mammalian telomeres has just started to be elucidated and through this extra views we highlight one aspect of that structure. We have recently identified the terminal nucleotides of both the C rich and G-rich telomere strands in human cells and showed that approximately 80% of the C-rich strands terminate precisely in ATC-5', while the last base of the G strand is less precise. This finding has important implications for the processing events that act on the telomere ends post-replication. While the mechanism behind this phenotype is yet to be unraveled, we discuss potential models that could explain the last base specificity. PMID- 16258280 TI - The mitotic checkpoint in cancer therapy. AB - The mitotic checkpoint is a key cell cycle control mechanism that ensures an accurate segregation of chromosomes during mitosis by delaying the onset of anaphase until all chromosomes are properly attached to a bipolar mitotic spindle. While complete loss of this checkpoint is lethal in vertebrates, a weakened mitotic checkpoint is frequently seen in cancer cells and it may contribute to tumorigenesis. Many anti-tumor drugs, including spindle assembly inhibitors and DNA damaging agents, can activate the mitotic checkpoint. However, since these drugs influence interphase events besides activating the mitotic checkpoint, the role of the mitotic checkpoint in drug-induced cell death remained unclear. Using a KSP antagonist that specifically acts on mitotic cells, we have recently shown that activation of the mitotic checkpoint followed by mitotic slippage or adaptation, activates Bax and initiates apoptosis. Notably, cells with a weakened mitotic checkpoint incur much less apoptotic death than their checkpoint-proficient counterparts, indicating the requirement of a competent mitotic checkpoint in the induction of apoptosis. In light of these findings and other recent reports, the potential influence of the mitotic checkpoint in response to chemotherapies, and the strategy to target the mitotic checkpoint for cancer therapeutics are discussed. PMID- 16258281 TI - Premeiotic origin of teratomas: is meiosis required for differentiation into mature tissues? AB - By virtue of meiotic cell division, primordial germ cells with heterozygous alleles develop into postmeiotic germ cells with homozygous alleles. Female and male germ cells may develop tumors--so-called teratomas--with a unique coexistence of a variety of histological elements from all three embryonic germ layers. In particular, mature teratomas consist exclusively of developmentally mature tissues whereas immature teratomas contain variable amounts of mature and immature tissues. In this study, we report genetic analysis of individual tissue components from mature and immature teratomas. The majority of mature teratomas showed consistent and concordant homozygous alleles in all selectively procured tissue components. In a small subset of mature teratomas, we observed discordant homozygous alleles. In contrast, immature teratomatous tissue revealed a heterozygous genotype. Remarkably, mature tissue components within immature teratoma revealed homozygosity. The findings suggest that immature teratomas and at least a subset of mature teratomas may originate from premeiotic cells, and implicate that meiosis may be required for differentiation into mature tissues. PMID- 16258282 TI - Cytokinesis and the spindle midzone. AB - At the end of the cell cycle a cell physically divides into two daughter cells in a process called cytokinesis. Cytokinesis consists of at least four steps: (1) The position of the presumptive cytokinesis furrow is specified. (2) A contractile ring is formed. (3) The contractile ring contracts, resulting in furrow ingression. (4) Cytokinesis completes with sealing of the membranes. The mitotic spindle positions the cytokinesis furrow at the cell cortex midway along the longitudinal axis of the spindle, which is both the mid-point between the two asters and the location of the spindle midzone. The mitotic spindle emits two consecutive signals that position the furrow: Microtubule asters provide a first signal; the spindle midzone provides a second signal. Our results support the view that the spindle midzone is dispensable for completion of cytokinesis. However, the spindle midzone can negatively affect aster-positioned cytokinesis, possibly because the aster- and midzone-positioned furrows compete for contractile elements. PMID- 16258284 TI - PTPRV is a key mediator of p53-induced cell cycle exit. AB - The p53 tumor suppressor functions as a sequence-specific DNA-binding transcription factor that promotes antiproliferative responses, including cell cycle checkpoints, cellular senescence and apoptosis. The precise nature of the p53 transcriptional programs and the complex mechanisms that govern whether or not a cell dies in response to p53 activation remain elusive. We have recently reported the identification of a new direct p53 target, Ptprv, encoding a transmembrane tyrosine phosphatase. Ptprv expression is dramatically and preferentially increased in cells undergoing p53-dependent cell cycle exit, but not in cells undergoing p53-mediated apoptosis. Importantly, while p53-induced apoptosis is intact in mice lacking Ptprv, Ptprv-null cells are defective in G1 checkpoint control. In addition, we report herein that Ptprv is induced at high cell density and mediates contact inhibition of cell growth. Together, the data suggest that Ptprv is a potent inhibitor of cell proliferation and a critical mediator of p53-induced cell cycle exit. PMID- 16258283 TI - From immature lamin to premature aging: molecular pathways and therapeutic opportunities. AB - Accelerated aging or progeria has been a puzzling disease for many years. The recent findings involving the lamin A/FACE-1 (substrate/protease) system in the etiology of Hutchinson-Gilford progeria syndrome and related pathologies have shed some light on the mechanisms underlying the development of these devastating conditions. Thus, genetic defects in the nuclear envelope protein prelamin A or in the FACE-1 metalloprotease (also called Zmspte24) involved in prelamin A proteolytic maturation, cause the accumulation of an abnormal form of this protein and the subsequent disruption of nuclear envelope integrity. Recently, we and others have observed how this disruption leads to alterations in chromatin organization, genomic instability, transcriptional changes, and activation of a p53-linked signaling pathway. By using genetic manipulation approaches in mouse, we have shown that lowering prelamin A levels results in a total recovery of Zmpste24-deficient mice from the accelerated aging process. Moreover, p53 nullizygosity allows a modest but significant improvement in the premature aging phenotype, and contributes to delaying the onset of the progeroid condition. On the basis of these results, we propose different potential therapeutic approaches that could be tested in Zmpste24-deficient mice. These strategies, some of which are based on existing drugs, might contribute to the development of effective treatments for these dramatic pathologies. PMID- 16258285 TI - The absence of p53 aggravates polyploidy and centrosome number abnormality induced by Aurora-C overexpression. AB - Aurora-C is the third member of the aurora serine/threonine kinase family and was found only in mammals. Because Aurora-C is overexpressed in many different types of cancer cells we decided to analyze the consequences of Aurora-C overexpression in human cells. We first investigated the subcellular localization of overexpressed GFP-Aurora-C in mitosis and interphase in HeLa cells. As expected, during mitosis, we found that Aurora-C mimics Aurora-B. Surprisingly, in few interphase cells, we found that Aurora-C localized to the centrosome, like Aurora A. We then examined the phenotype generated by Aurora-C overexpression. Basically it looked similar to the phenotypes observed after overexpression of the other Aurora kinases. We observed an augmentation of polyploid cells containing more than two centrosomes. More interestingly this phenotype was aggravated in the absence of a functional p53. Although the physiological function of Aurora-C in somatic cells remains to be clarified, our results, just like for the two other Aurora kinases, raised the question of a role of Aurora-C in the development and progression of cancer especially in the presence of mutated p53. PMID- 16258286 TI - CDK-dependent stabilization of Cdc6: linking growth and stress signals to activation of DNA replication. AB - Cyclin-dependent kinases (CDKs) play a crucial role in cell cycle progression by controlling the transition from G(1) phase into S phase where DNA is replicated. Key to this transition is the regulation of initiation of DNA replication at replication origins. CDKs are thought to regulate origins of replication both positively and negatively by phosphorylating replication proteins at origins. Several replication proteins that are potentially negatively regulated upon CDK phosphorylation have been identified. However, the mechanism by which CDKs activate replication is currently less well understood. New observations revealing that the initiation protein Cdc6 is stabilized by CDK2-dependent phosphorylation may give more insight in this process. PMID- 16258287 TI - Stathmin prevents the transition from a normal to an endomitotic cell cycle during megakaryocytic differentiation. AB - Physiological polyploidy is a characteristic of several cell types including the megakaryocytes (MK) that give rise to circulating blood platelets. MK achieve polyploidy by switching from a normal to an endomitotic cell cycle characterized by the absence of late mitotic stages. During an endomitotic cycle, the cells enter into mitosis and proceed normally through metaphase and early anaphase. However, late anaphase, telophase and cytokinesis are aborted. This abortive mitosis is associated with atypical multipolar mitotic spindles and limited chromosome segregation. Stathmin is a microtubule-depolymerizing protein that is important for the regulation of the mitotic spindle and interfering with its expression disrupts the normal mitotic spindle and leads to aberrant mitotic exit. As cells enter mitosis, the microtubule depolymerizing-activity of stathmin is switched-off, allowing microtubules to polymerize and assemble into a mitotic spindle. Reactivation of stathmin in the later stages of mitosis is necessary for the disassembly of the mitotic spindle and the exit from mitosis. Previous studies had shown that stathmin expression is downregulated as MK become polyploid and inhibition of its expression in K562 cells increases their propensity to become polyploid. In this report, we describe our studies of the mechanism by which stathmin plays its role in MK polyploidization. We show that stathmin overexpression prevents the transition from a mitotic cycle to an endomitotic cycle as determined by a decrease in the number of multipolar mitotic spindles. These observations support a model in which downregulation of stathmin expression in megakaryocytes and other polyploid cells may be a critically important factor in endomitosis and polyploidy. PMID- 16258288 TI - Thrombopoietin stimulates vascular endothelial cell growth factor (VEGF) production in hematopoietic stem cells. AB - Thrombopoietin (TPO) is a pivotal and nonredundant hematopoietic cytokine, supporting the survival, self-renewal activity and proliferation of hematopoietic stem and progenitor cells, the growth and differentiation of megakaryocytes, and the functional activation of their progeny, blood platelets. TPO exerts these effects through regulating the abundance or subcellular localization of several transcription factors, including the homeodomain proteins HOXB4 and HOXA9. In addition to these effects, TPO helps orchestrate a cytokine-network in the bone marrow microenvironment that supports hematopoietic stem cell (HSC) function. In recent studies we have shown that TPO stimulates production of vascular endothelial cell growth factor (VEGF), another cytokine vital for HSC physiology, promoting their survival and expansion into committed hematopoietic progenitors. Like several other effects of the cytokine, the effect of TPO on VEGF expression is mediated by stabilization and activation of the primary transcription factor responsible for VEGF expression, the oxygen tension responsive hypoxia inducible factor-1 (HIF-1). Together with the observation that bone marrow microenvironment is hypoxic and hypoxia simulates the repopulating activity of HSCs, our observations suggest that TPO mimics hypoxia and controls important genes required for HSC cycling, including VEGF, adding to our understanding of how the hormone contributes to HSC function. PMID- 16258289 TI - Individual adult human neurons display aneuploidy: detection by fluorescence in situ hybridization and single neuron PCR. AB - Neurons, once committed, exit the cell cycle and undergo maturation that promote specialized activity and are believed to operate upon a stable genome. We used fluorescence in situ hybridization, selective cell microdissection, and loss of heterozygosity analysis to assess degree of aneuploidy in patients with a neurodegenerative disease and in normal controls. We found that aneuploidy occurs in approximately 40% of mature, adult human neurons in health or disease and may be a physiological mechanism that maintains neuronal fate and function; it does not appear to be an unstable state. The fact that neuronal stem cells can be identified in adult humans and that somatic mosaicism may be found in neuronal precursor cells deserves further investigation before using adult neural stem cells to treat human disease. PMID- 16258290 TI - A role for G-CSF (granulocyte-colony stimulating factor) in the central nervous system. AB - G-CSF (Granulocyte-colony stimulating factor) is a hematopoietic growth factor that has been known for 20 years, and has been named for its role in the proliferation and differentiation of cells of the myeloic lineage. We have uncovered a novel spectrum of activities of G-CSF in the central nervous system. G-CSF and its receptor are expressed by neurons in many brain regions, and are upregulated upon experimental stroke. In neurons, G-CSF acts anti-apoptotically by activating several protective pathways. In vivo, G-CSF decreases infarct volumes in acute stroke models in rodents. Moreover, G-CSF stimulates neuronal differentiation of adult neural stem cells in the brain, and improves long-term recovery in more chronic stroke models. Thus, G-CSF is a novel neurotrophic factor, and a highly attractive candidate for the treatment of neurodegenerative conditions. Here we discuss this new property of G-CSF in contrast to its known functions in the hematopoietic system, summarize data from other groups on G CSF's actions in cerebral ischemia, compare G-CSF to Erythropoietin (EPO) in the CNS, and highlight clinical implications. PMID- 16258291 TI - Effects of reduction mammoplasty on upper-limb nerve conduction across the thoracic outlet in women with macromastia: a prospective randomized study. AB - Macromastia is a common problem. The physical complaints include upper body pain and aches. There have been anecdotal reports of neurologic deficits in the nerves emanating from the lower trunk of the brachial plexus. This is thought to be due to pressure on the lower trunk from both the first rib and tilting forward of the coracoid process. Other anecdotal reports have centered on the correction of neuropraxia of the ulnar nerve following bilateral breast reduction (BBR). We investigate the effect of BBR on the electrophysiological function of the nerve supply to the upper limbs in women with macromastia. METHODOLOGY: Consecutive patients undergoing BBR were randomized into 2 groups, depending on time of surgery. None had any prior neurologic disorder. Each patient had a comprehensive neurologic assessment and 2 electrodiagnostic neurophysiologic tests. Group 1 had 2 tests, one before surgery and a second 3 months postsurgery, while Group 2 had 2 sets of tests, one initially and a second test 4 months later (control). The outcome measures include somatosensory evoked potential (SSEP) (median and ulnar), F-wave median and ulnar latencies. The F waves measure the integrity of neural conduction time from the anterior horn cells to the hypothenar and thenar muscles reflecting lower trunk function. The SSEP of the median nerve measure the integrity of the nerve fibers traversing the upper trunk of the brachial plexus and the ulnar nerve SSEP that of the lower trunk. There was no statistical difference in the conduction times. BBR does not have any effect on the upper limb nerve conduction times. PMID- 16258292 TI - Muscle activity in the partially paralyzed face after placement of a fascial sling: a preliminary report. AB - Neuromuscular re-education (ie, physical therapy) is often the first treatment given to patients with a partial facial paralysis. The purpose of this paper was to examine whether by repositioning and supporting partially paretic muscles with a fascial sling, one could improve facial movement in patients for whom the benefits of physical therapy had plateaued. Six patients with a history of unilateral, partial facial paralysis were assessed using the Facial Grading System (FGS) and surface electromyography (EMG) recordings of facial muscle activity. Automated facial analysis (AFA) was used to measure the facial excursions of the most recent patient. The FGS composite scores indicated improvement following static sling placement in all patients. The FGS subscale scores for voluntary movement indicated that the excursion of facial movement increased in 4 of the 6 patients. Surface EMG data demonstrated increased muscle activity in the zygomaticus major muscle in all patients. AFA demonstrated that following sling placement, the excursion of the lip commissure nearly doubled. The sling procedure, traditionally considered an intervention for improving static symmetry of the face, may also be useful for enhancing movement in some patients with a partial facial paralysis. Additional data, such as measurements provided by AFA, are needed to correlate facial displacement with EMG muscle activity. PMID- 16258294 TI - Techniques of external monitoring of buried free flaps. AB - Postoperative monitoring of free tissue transfer has proven to be crucial in enhancing the flap salvage rate in the event of thrombosis of the anastomosed vessels. Unfortunately, for buried flaps in the head and neck, direct monitoring of the flap can be extremely difficult, if not impossible. Utilizing various methods of flap exposure in a series of 55 buried free flaps of the head and neck, we demonstrate that we were able to provide a reliable means of direct postoperative assessment of the free flaps. Exteriorized components (indicator flaps) were established for 52 cases. An incision to create a skin window in the neck for monitoring was used for 3 cases. For the exteriorized flaps, in addition to clinical observation, adjunctive continuous monitoring using laser Doppler flowmetry was employed for 4-5 days. One free flap developed vascular thrombosis, which was re-explored and salvaged, giving a flap success rate of 100%. Fifty of 52 (96.2%) external components remained viable and therefore reliable for monitoring the main body of the flap. Two indicator flaps were unreliable after postoperative day 1 due to poor skin perfusion, while the subcutaneous component clinically remained viable. There was no return to the operating room for false positive reexploration. This series reinforces the feasibility and reliability of direct monitoring of buried free flaps using the laser Doppler in practically all cases when modification is added to the flap design and inset. PMID- 16258293 TI - Repair of large nasal septal perforations using facial artery musculomucosal (FAMM) flap. AB - BACKGROUND: Nasal septum perforations from surgical submucous resection, septoplasty, blunt trauma, and substance abuse may cause epistaxis, nasal obstruction, discharge, crusting, dryness, pain, and whistling. While small symptomatic perforations are closed with local mucosal flaps, options for closure of large symptomatic perforations are limited. A local pedicled flap, the facial artery musculomucosal (FAMM) flap was studied in patients with large symptomatic nasal septal defects. METHODS: Patients included in the study had (1) a nasal septal defect measuring at least 20 mm in greatest dimension; and (2) related symptoms of nasal crusting, discharge, dryness, obstruction, epistaxis, pain, or whistling. Six patients (3 males; 3 females) met these criteria and received FAMM flap repair. Outcomes were assessed based on comparison of preoperative versus last follow-up (range, 10-30 months; mean 17 months) assessment of perforation size and symptomatology. Overall discomfort was rated at each time point on a 1 10 scale. RESULTS: Age at time of operation ranged from 21 to 44 years, with a mean of 34 years of age. Causes of septal perforation included blunt trauma (50%), cocaine abuse (33%), and submucous resection (17%). Preoperatively, maximal recorded dimensions of septal perforations ranged from 3.1 to 4.0 cm with a mean of 3.5 +/- 0.4 cm. Symptoms included pain (83%), dryness (67%), crusting (50%), discharge (33%), epistaxis (33%), and obstruction (33%). Three or more symptoms were experienced by 5 patients (83%). Overall discomfort ranged from 6 10, with a mean of 8.4. Postoperatively at last follow-up, all 6 patients (100%) achieved closure of their septal defect (P < 0.001). Overall discomfort score was zero for all 6 patients (100%) (P < 0.0001). Complete symptomatic resolution was also noted among all 6 patients (100%) (P < 0.01). CONCLUSIONS: In summary, the advantages of the FAMM flap closure technique were (1) no visible external scar, with minimal donor site morbidity; (2) successful closure of large septal defects (>2 cm) with vascularized tissue in a single stage; and (3) resolution of patient symptomatology. PMID- 16258295 TI - The proximal pedicled anterolateral thigh flap for lower limb coverage. AB - Although primarily considered as a versatile free-flap donor site, the anterolateral thigh can also be a source of a local muscle perforator flap. This attribute has previously been rarely considered for lower limb coverage. This small series of 3 additional cases demonstrates the usefulness of a proximal pedicled anterolateral thigh flap for medial and lateral thigh wounds. This flap can also be part of a combined flap, in particular when transferred with the vastus lateralis muscle as a local chimeric flap. The peninsular version of the anterolateral thigh local flap avoids venous congestion and is very reliable. The orthograde pedicled anterolateral thigh muscle perforator flap should be considered as another useful alternative for any upper thigh wound if a flap is essential. PMID- 16258296 TI - Formation of functional neovagina with vertical rectus abdominis musculocutaneous (VRAM) flap after total pelvic exenteration. AB - BACKGROUND: Pelvic exenteration may be the only curative option for women with recurrent pelvic malignancies. After total pelvic exenteration, the resultant perineal defect heals slowly if left to do so by secondary intention. Reconstruction with the vertical rectus abdominis musculocutaneous (VRAM) flap brings a generous bulk of healthy tissue into the defect, speeding recovery by facilitating primary healing. METHODS: Six women underwent reconstruction of a neovagina using a vertical rectus abdominis musculocutaneous flap. All 6 had total pelvic exenteration for advanced gynecologic malignancy. Primary diagnosis was cervical carcinoma (n = 3), vulvar carcinoma (n = 1), nonsmall cell vaginal cancer (n = 1), and vaginal melanoma (n = 1). Four patients had received adjuvant radiotherapy preoperatively. RESULTS: All flaps remained 100% viable postoperatively. There were no cases of fistula, infection, or bowel obstruction. Two patients died of cardiovascular arrest postoperatively. The 4 other patients report satisfaction with reconstruction. Three had vaginal intercourse with orgasm. CONCLUSION: The inferiorly based vertical rectus abdominis musculocutaneous flap is a dependable source of tissue for pelvic reconstruction and is the flap of choice in the Division of Plastic Surgery. In addition to facilitating healing, the VRAM flap (neovagina) improves a woman's psychosocial well-being. PMID- 16258297 TI - Through-and-through gunshot wounds to the foot: the "Fearless Fosdick" injury. AB - Through-and-through gunshot wounds of the foot present the unique problem of needing to fill a defect while at the same time providing coverage of the dorsum and plantar surface of the foot. A series of 5 patients from 2 institutions is presented. These patients all sustained gunshot wounds that penetrated the forefoot, leaving a rim of uninjured soft tissue and bone around the periphery. All patients were reconstructed with free-tissue transfers, but no bone grafting was required to replace missing bone. One patient did have an arthrodesis and wedge osteotomy to help with walking mechanics. Soft-tissue reconstruction only is adequate to restore the foot sufficiently to allow the patient to walk satisfactorily. PMID- 16258298 TI - Use of fascia component of the anterolateral thigh flap for different reconstructive purposes. AB - The anterolateral thigh flap is commonly used for reconstruction of soft-tissue defects located at various sites of the body. This versatile flap offers many advantages to the reconstructive microsurgeons for the treatment of difficult defects. From 2000 to 2005, 70 anterolateral thigh flaps were transferred to reconstruct soft-tissue defects. We retrospectively reviewed these patients and found that the fascia lata component of the flap was used for different purposes in 19 patients. The fascia lata component of the flap was used for suspension of the flap in lip reconstruction in 12 patients, for reconstruction of dural defect in the scalp in 2 patients, for reconstruction of tendon defects in the forearm in 3 patients, and for reconstruction of fascia defect in the abdominal wall in the remaining 2. Complete loss of the flap was seen in an anterolateral thigh flap (5.2%) that was used for lower lip reconstruction. One flap necrosed partially (5.2%), and it was treated with surgical debridement and transposition of latissimus dorsi musculocutaneous flap. The objective of this study is to focus on the reliability of the fascial component of the anterolateral thigh flap. Although many authors have described other advantages of the anterolateral thigh flap extensively, this peculiarity has not been stressed adequately. Anterolateral thigh flap offers a thick and vascular fascial component with large amounts that can be used for different reconstructive purposes, and it should be taken into consideration as an important advantage of the flap, together with other well-known advantages. PMID- 16258299 TI - Biobrane in the treatment of burn and scald injuries in children. AB - Eighty-four children with a burn (n = 7) or scald injury (n = 77), treated with Biobrane, were investigated in a retrospective and clinical study. In most patients (n = 71), the Biobrane was adherent and without any reactions or infections. An infection was seen in 10.7% (9 from 84 patients). Twenty-one of 49 patients of the follow-up had limited scar areas. In scarless healed areas, 54.3% had normopigmentation and 39.1% were hypopigmented. The skin quality of the scars was mostly hypopigmented with a softness between minimal and middle resistance, under 2-mm high, and of normal to pink skin color. Compared with other dressings, Biobrane is no more expensive than others. We conclude that when used on properly selected wounds, Biobrane is an effective and, for the children, less traumatic therapy for superficial partial-thickness burns without increasing the cost of outpatient burn care. PMID- 16258300 TI - Evaluation of antiseptic use in plastic and hand surgery. AB - Despite the growing complexity of plastic surgical cases, antiseptics available for preoperative preparation do not always prove effective. In addition to the varied sites of plastic surgery, there is a likelihood of considerable contamination with microorganisms since surgical gloves are frequently punctured during surgical procedures. With an aim to find a solution to all of these problems, a study was conducted in 2 stages. First, antimicrobial effects on the normal flora of hands of 4 different surgical handwashing procedures, which consist of scrubbing with 7.5% povidone-iodine (PI) scrub by using the sponge/brush, 7.5% PI scrub alone, 0.5% benzalkonium chloride (BC) solution, and 0.1% BC solution, were compared. In the second stage, bacteriological samplings were obtained from fingertips of the operating team during the surgical procedure. At the end of the study, the following results were obtained: (1) scrubbing with 7.5% PI detergent by using a scrub sponge was significantly more effective than without using a scrub sponge (P < 0.05). (2) There was no statistically significant difference in scrubbing when povidone-iodine detergent and 0.5% benzalkonium chloride solution (P > 0.05) were compared. (3) At the end of the surgery lasting 1-5 hours, hands were found to be cleaner than previously washed hands for preoperative preparation, which was considered a consequence of the sustained effect of PI. (4) There was a statistically significant difference between the cultures taken from the gloves at the end of the operation and from the hands after the gloves were taken off (P < 0.01). At the end of the operation, the gloves themselves were found to be extremely dirtier than the hands inside them. As a conclusion, bacterial escape due to glove perforation occurs from outside of the glove to the inside, contrary to the generally accepted concept. PMID- 16258301 TI - Anthropometric proportion indices in the craniofacial regions of 73 patients with forms of isolated coronal synostosis. AB - The study group consisted of 73 North American patients (29 males and 44 females), aged between 0-5 months and 20 years, 26 with right, 33 with left, and 14 with bicoronal synostosis. Basic proportion indices were established in 5 craniofacial regions (cranial, facial, orbital, nasal, oral) calculated from 2 projective measurements [cranial: eu-eu, g-op; facial: n-gn, zy-zy; orbital: en en, ex-ex; nasal: al-al, n-sn; oral: sn-sto, ch-ch (eu, eurion; g, glabella; op, opisthocranion; n, nasion; gn, gnathion; zy, zygion; en, endocanthion; ex, exocanthion; al, alare; sn, subnasale; sto, stomion; ch, chelion)] taken from the patients by the first author before surgery. These data were then compared with the anthropometric norms established for North American whites (Farkas LG. Anthropometry of the Head and Face. 2nd ed. Raven Press; 1994). In males, the total percentage of normal, subnormal, and supernormal proportion indices was smaller than in females in all 3 groups of patients. Generally, the difference in percentage between normal proportions was the smallest (13.1%) and the supernormal one the greatest (68%). In males, the oral measurements showed the highest frequency of normal proportions (100%) in all 3 patient groups, and the cephalic index the lowest (60%-66.7%). Among the abnormal proportions, the supernormal ones were found more often, especially in the cranial region of females with bilateral coronal synostosis (66.7%). Subnormal proportion indices were rare: none were seen in the facial region of males. A subnormal cephalic index (eu-eu/g-op) was not present in either sex in right and bicoronal synostosis and was observed in only 2 males with left coronal synostosis (13.3%). The study showed that the influence of synostosis gradually decreased from the top to the bottom of the face, with the oral region showing no abnormalities. PMID- 16258303 TI - Rat penis as a replantation model. AB - The aim of this study is to assess the feasibility of rat penile replantation as a new microsurgical training model. The study was performed in 2 parts. ANATOMIC STUDIES: Fifteen Wistar albino rats were used to study and document the penile vascular anatomy. In 5 rats, dissections were performed after colored silicone injections, while 5 rats were operated under anesthesia to develop the strategy of flap elevation. In the remaining 5 rats, microangiographic study was performed with silicone-lead oxide mixture. FLAP STUDIES: As flap studies in 5 rats, penis were elevated based on right-side internal pudendal artery and internal pudendal vein and resutured. In 6 rats, penis were elevated as free flaps, and in 3 rats the penis were implanted in right thigh of the rats being the femoral artery and vein recipient. In the remaining 3 rats, penis were resutured in their original place, with saphenous artery and vein being the recipient and rerouted to the pubic region. At postoperative fifth day, the penis were examined for viability, and selected ones were histologically examined. Rat penis has a dual blood supply from bilateral internal pudendal arteries. Venous drainage is via both crural veins and dorsal vein. One side of the internal pudendal artery and anastomotic vein (branch of pudendal plexus) may be used as the vascular pedicle of the flap. Rat penis may be successfully elevated as a free flap and also may be replanted in its original place. PMID- 16258302 TI - Use of the artificial dermis for free radial forearm flap donor site. AB - The radial forearm free flap has been popular in many area of reconstructive surgery. Despite the many attributes of this flap in surgical reconstruction, one of the clear disadvantages has been the morbidity of the donor site. To solve problems associated with closure of the radial forearm flap donor site, the artificial dermis was used for better outcome in the donor site. From July of 2001 to October 2003, 13 patients (12 male, 1 female) ranging in age from 27 to 67 years underwent free radial forearm transfers. Flap size measured 49 to 144 cm2. Donor-site defects secondary to the flap elevation was closed first with the artificial dermis (Terudermis), marketed by Terumo Inc, Japan, and secondary split-thickness skin graft to the artificial dermis was performed in 15 days (range, 13 to 17 days) on average. There was no flap failure in all cases. Partial loss of the artificial dermis graft was noticed in 2 cases, but it was negligible. Secondary skin graft take was noticed in about 7 days, and there was no remarkable skin graft loss. The artificial dermis was used successfully in 13 patients with the donor defects of free radial forearm flap. Despite the disadvantage to need the secondary split-thickness skin graft, use of the artificial dermis gave us not only soft-tissue augmentation and graft-skin quality similar to full-thickness skin graft but also fast graft healing and diminished morbidity of the skin-graft donor site. PMID- 16258304 TI - Thrombotic effect of purposeful back-wall stitch for different suture locations and vessels in rats. AB - Many microsurgeons experience inadvertent back-wall stitch as a cause of immediate anastomotic failure. To investigate this factor as a possible cause of delayed arterial anastomotic failure that does not appear in the operation room, a purposeful back-wall stitch (PBWS) model of microarterial anastomosis was applied in various configurations on femoral and carotid arteries of rats. Carotid (n = 28) and femoral (n = 28) artery groups were equally divided into 4 different subgroups according to the type of PBWS. Control subgroups had no PBWS. Thirty-degree, 60-degree, and 90-degree subgroups had PBWS located at 30, 60, and 90 degrees, respectively. Patencies were assessed at 60 minutes and 24 hours. Immediate milking test demonstrated normal anterograde refilling in all anastomoses. PBWS increased thrombosis in femoral arteries, while it did not increase it in the carotid at either evaluation times. The only significant difference was between 90 degrees PBWS and the control subgroup irrespective of the vessel factor in 24 hours. Histologic examination supported absence of thrombosis, as suggested by surgical examination. This may show that small-sized arteries are more inclined to thrombosis compared with larger ones and the thrombosis risk increases as the inadvertent back-wall stitch is more centrally located. Contrary to general notion that the inadvertent back-wall stitch causes immediate thrombosis, thromboses later than 1 hour, and even patent anastomoses in 24 hours, were observed in femoral and carotid artery groups. This study suggests that the inadvertent back-wall stitch should also be considered as a possible cause of late anastomotic problems. PMID- 16258305 TI - Combined treatment with CD40 costimulation blockade, T-cell depletion, low-dose irradiation, and donor bone marrow transfusion in limb allograft survival. AB - To determine the efficacy of a regimen based on CD40 costimulation blockade and donor bone marrow in the limb allograft model, C57Bl/6 mice received limb allografts from Balb/c mice and either no treatment or a combination of MR1 (anti CD40 ligand monoclonal antibody), CD4+ and CD8+ T-cell-depleting antibodies, low dose irradiation, and bone marrow transfusion from Balb/c donors for 1 or 2 weeks. Recipients treated for 1 week showed rejection at 38.2 +/- 5.4 (mean +/- SEM) days, while those treated for 2 weeks had allograft survival of 56.5 +/- 9.9, with a range up to 91 days. Histology demonstrated rejection which was less cell-mediated and suggestive of transplant vasculopathy. Differential rejection of skin occurred first. Thus, a combined regimen based on CD40 costimulatory blockade and donor marrow significantly prolonged allograft survival. However, tolerance was not achieved, and histology suggests chronic rejection as a possible cause of allograft loss. PMID- 16258306 TI - Mandibular distraction in a neonate with muscular dystrophy. AB - The following case report describes the successful use of distraction osteogenesis (DO) for the treatment of hypoplasia of the mandible in a patient with muscular dystrophy (MD). While DO has been used for hypoplasia of the mandible, no evidence exists that is it safe in the setting of MD. MD is a disease that primarily affects skeletal muscle; however, pathologic changes in the adjacent bone have been described. Furthermore, the healing of involved bone may be problematic, making DO a potentially unsuccessful technique in this group of patients. This report is the first successful utilization of DO for the treatment of mandibular hypoplasia in a patient with MD. PMID- 16258307 TI - Giant pleomorphic adenoma of the parotid gland with malignant transformation. AB - A 72-year-old female is described who was admitted to the hospital with a giant mixed tumor of the left parotid gland that had grown to a weight of 6051 g within 20 years and ultimately underwent resection of the mass. Histopathological examination revealed adenocarcinoma cells with hyperchromatic nuclei and mitotic figures proliferated in pleomorphic adenoma. Many authors have reported giant tumors weighing more than a few kilograms of the parotid glands, and most were benign. The present tumor was the sixth largest among all the parotid tumors, including both benign and malignant, which were reported. Furthermore, our case was the largest one confirmed as malignant recorded in the world literature. The tumor was successfully resected with no marked facial nerve paralysis. PMID- 16258308 TI - Novel technique for venous augmentation in a free deep inferior epigastric perforator flap. AB - Venous congestion in a free deep inferior epigastric perforator flap threatens the viability of the flap and can lead to eventual flap loss. We describe a novel technique for flap salvage by anastomosing the ipsilateral superficial inferior epigastric vein to a venae comitantes of the deep inferior epigastric pedicle. When recognized intraoperatively, venous congestion can be relieved immediately without the need for additional dissection of recipient vessels. This technique can also be used during reexploration for flap congestion. We routinely preserve length on the superficial inferior epigastric vein for potential flap salvage. PMID- 16258309 TI - Salvage treatment of an irradiated, infected lumbosacral wound. AB - A 60-year-old male with lumbosacral multiple myeloma received 5100 cGy of external-beam radiation, thalidomide, and Decadron. He subsequently underwent excision of the epidural tumor, decompressive L4, L5, and S1 laminectomies, and bilateral L4-5 and L5-S1 medial facetectomies. The patient developed osteoradionecrosis, cerebrospinal fluid leak, wound infection, and sepsis. Debridement and bilateral V-Y fasciocutaneous advancement flaps failed. Pedicled omental transposition flap through a Petit triangle tunnel was successfully performed. Omental transposition provides a safe option for salvage treatment of irradiated, infected lumbosacral wounds. The plastic and trophic qualities of the omentum make it an excellent choice to fill poorly vascularized wounds. In addition to its immunologic and neoangiogenic properties, the omentum has a dense lymphatic network with tremendous absorptive potential. Its biologic advantages must be weighed against the need for celiotomy and available local options according to circumstances. PMID- 16258310 TI - Revisiting vascularized muscle flaps for complicated sternal wounds in children. AB - Surgeons at our center previously reported a case of a 2-month-old infant who underwent closure of an infected sternal wound following open cardiac surgery with debridement followed by closure with bilateral pectoralis muscle flaps and a unilateral rectus abdominis muscle flap. The success of the procedure has spawned refinements in the technique, such as the one described herein. A 2-week-old neonate was evaluated for postoperative sternal dehiscence and instability following open cardiac surgery for severe congenital cardiac anomalies. Management included initial debridement and application of a vacuum-assisted closure (V.A.C.) system (KCI, Oxfordshire, UK). In conjunction with the final V.A.C. dressing change, the patient underwent delay of the left rectus muscle by division of the inferior epigastric pedicle. She subsequently underwent transposition of the left rectus muscle and application of a full-thickness skin graft for coverage of the sternal defect. She has since done well and still requires further invasive cardiac procedures. PMID- 16258312 TI - Double-strand suturing fixation technique for treatment of acute volar plate avulsion fracture of the base of the middle phalanx. AB - A new suturing technique for treatment of acute volar plate avulsion fracture of the proximal interphalangeal joint is reported. Because this technique that consists of double-strand suture is rigid enough and never obstructs joint motion, early motion exercise can be allowed. This technique only demands simple skill like conventional K-wire fixation. Furthermore, removal of fixation material is not necessary. PMID- 16258311 TI - Superficial dorsal artery of the forearm: case report and review of the literature. AB - Although abnormalities of vascular anatomy in the forearm are common, variations of the radial and ulnar arteries are rare. Nevertheless, arterial variants in the forearm may present clinically with neurovascular signs or symptoms. Even when anomalous arteries are not apparent, they may complicate surgery of the forearm and hand, as well as reconstructive surgery that involves the harvest of radial or ulnar artery-based forearm free flaps. For example, the superficial ulnar artery has an incidence of 2.7% and has been well described as a "hidden trap" in the harvest of radial forearm flaps. We present a case report of a variant of the radial artery-the superficial dorsal artery of the forearm. This artery has an estimated incidence of 0.75% and is associated with either an absent or small caliber radial artery. We believe this is the first report of such an artery presenting to clinical attention, as well as the first imaging of this structure with magnetic resonance angiography. The embryology of this structure and its clinical implications to the hand surgeon, peripheral vascular surgeon, and reconstructive microsurgeon, are also discussed. PMID- 16258314 TI - A scalpel in the wrong hands. PMID- 16258313 TI - Strategies for tolerance induction in nonhuman primates. AB - Recent advances in the field of reconstructive surgery and immunology resulted in increased interest in composite tissue allograft (CTA) transplantation. Up to date, more than 50 CTA transplants have been reported in humans. A significant number of experimental studies on CTA transplants under different protocols of tolerance-inducting strategies have been reported in small-animal models. There is however, a limited number of CTA transplants performed in nonhuman primates. To reach the ultimate clinical success in CTA transplantation, more experimental studies on tolerance induction in nonhuman primates are needed to apply these immunomodulatory protocols to CTA transplants in humans. In this review, strategies for tolerance induction in the nonhuman primate model in solid organ and CTA transplants are presented in 3 major categories: chimerism induction, T cell depletion, and costimulatory receptor blockade. PMID- 16258315 TI - Hand surgical assistance: light and suction. PMID- 16258316 TI - Use of innovative hand vise grip in craniofacial surgeries. PMID- 16258317 TI - Chondrodermatis nodularis helicis: a simple adjunct to a nonsurgical approach. PMID- 16258319 TI - Role of new antifungal agents in prophylaxis of mycoses in high risk patients. AB - PURPOSE OF REVIEW: For pancreas, liver, and hematopoietic stem cell transplant recipients, no antifungal prophylaxis led to a high rate of and high morbidity from fungal infection. With the use of fluconazole as prophylaxis since the early 1990s, there have been shifts in the types of infecting fungal pathogens, documentation of resistance among fungal organisms, and changes in transplant practices. The aim of this article is to review recent clinical trials regarding antifungal chemoprophylaxis among several populations of high risk patients. RECENT FINDINGS: Itraconazole, micafungin, and posaconazole have been studied as alternatives to fluconazole prophylaxis. Itraconazole showed no dramatic improvement over fluconazole as prophylaxis during liver and hematopoietic stem cell transplantation, primarily due to gastrointestinal side effects. In addition, detrimental changes to cyclophosphamide metabolism were noted for hematopoietic stem cell transplant recipients. Micafungin was superior to fluconazole during the pre-engraftment period of hematopoietic stem cell transplantation, because it was able to prevent mold infections, required less switches to empirical antifungal therapy, and functioned as well as fluconazole in preventing yeast infections. Posaconazole was compared to fluconazole during a 16-week prophylaxis period during graft-versus-host disease, but results of this study are still forthcoming. Aerosolized amphotericin products appear to be safe for lung transplant recipients. SUMMARY: Fluconazole remains the standard agent for prophylaxis against invasive fungal infections for pancreas, liver, and hematopoietic stem cell transplant recipients. Micafungin is superior to fluconazole with minimal toxicity for use in the pre-engraftment period of hematopoietic stem cell transplantation. The optimal agent for prophylaxis later following transplant, if mold coverage is desired during prolonged immunosuppression, has not been determined. PMID- 16258320 TI - New antifungal agents under development in children and neonates. AB - PURPOSE OF REVIEW: This review focuses on only the newest antifungal agents recently approved or still under development and the available data in pediatric and neonatal patients. The larger body of data in adult patients is used for comparative purposes only in an attempt to understand pediatric implications. RECENT FINDINGS: Pharmacokinetic data suggest differences in dosing for many newer agents in children versus adult patients, but each agent has not been fully evaluated. Voriconazole displays non-linear pharmacokinetics in adults but has linear pharmacokinetics in children, necessitating a higher dose in smaller patients and potential treatment failures using the approved adult dosing schedule. Caspofungin likewise requires higher doses relative to adult patients, and dosing in children is best accomplished on a body surface area scheme and not a body weight dosing platform. Preliminary data suggest posaconazole, an investigational triazole, in children may lead to similar levels as in adults, but very limited efficacy data are available at any dose. Micafungin dosing has been explored in neonatal patients and there is a clear trend toward lower levels obtained in the very smallest infants, highlighting the importance of the neonatal period as a separate entity to even the pediatric age group. SUMMARY: Initial data suggest dosing differences in children with some antifungals, and other newer agents have not been fully tested for the correct dosing. The underlying concern of efficacy in children compared with adult patients has never been answered as there are no randomized, phase III antifungal clinical trials from which pediatric-specific data were obtained. PMID- 16258321 TI - Treatment of Candida infection: a view from the trenches! AB - PURPOSE OF REVIEW: To provide an overview of the treatment of Candida infections from the perspective of the infectious disease clinician in the community. An insight is given into relevant issues as they apply to clinical practice. As community hospitals strive to provide state-of-the-art medical care to critically ill patients, effective infection control programs and the infectious disease consultation availability become indispensable. RECENT FINDINGS: Candida infections rank as the fourth most common cause of nosocomial bloodstream infections. Although C. albicans remains the most common pathogen, emerging trends in invasive candidiasis are notable for a dramatic increase in infections due to non-albicans Candida species, reflecting changes in clinical practice. These trends affect our practice because infections with non-albicans species raise concerns for inherent decreased antifungal susceptibility, ultimately impacting our preventive, empirical and therapeutic approaches. It seems inevitable for most acutely ill patients today to be admitted to the intensive care unit, thus increasing their risk for nosocomial infections. In addition, cost containment efforts may force patients receiving intravenous therapy into less supervised environments, potentially increasing their risk for candidemia. SUMMARY: Potentially fatal Candida infections are commonplace in seriously ill hospitalized patients. The infectious disease physician is challenged to develop expertise in using the newly introduced antifungal agents, applying evidence based guidelines developed from quality randomized clinical trials. The infectious disease specialist is called to play a multidisciplinary role formulating infection control policies, developing drug formularies, educating the staff and treating the sickest patients. Infection surveillance in local communities coupled with available antifungal agents may improve our management of Candida infections. PMID- 16258322 TI - Treatment approaches for community-acquired methicillin-resistant Staphylococcus aureus infections. AB - PURPOSE OF REVIEW: This review addresses therapeutic approaches to community acquired methicillin-resistant Staphylococcus aureus (MRSA) infections, focusing on recently published data in the English-language medical literature dating from June 2004 to July 2005. RECENT FINDINGS: During the reviewed time period, the overall understanding of the epidemiology and virulence of community-acquired MRSA has continued to advance. This same period has also seen numerous works dealing with the newer and emerging anti-staphylococcal antimicrobial agents. Important clinical trials involving linezolid, daptomycin, tigecycline, dalbavancin, and telavancin have been completed. At the same time, owing to the cost of newer agents and the broad susceptibility pattern of community-acquired MRSA, many older antimicrobial agents (long-acting tetracyclines, fluoroquinolones, rifampin, trimethoprim-sulfamethoxazole, and clindamycin) have also been reexamined. SUMMARY: As data accumulates, the newer antimicrobial agents active against community-acquired MRSA continue to demonstrate their value. Despite the appeal and abundant publications involving newer agents, older antimicrobials certainly retain a therapeutic role. Considerable work needs to be done prospectively evaluating older agents for community-acquired MRSA disease. The most important therapeutic intervention for the majority of community acquired MRSA infections is adequate drainage of purulent fluid collections. Antimicrobial selection for community-acquired MRSA infections should be governed by disease severity, susceptibility patterns (especially based on timely clinical specimen culture), clinical response to therapy, and cost. PMID- 16258323 TI - Treatment of multidrug resistant Acinetobacter. AB - PURPOSE OF REVIEW: Acinetobacter baumannii-calcoaceticus complex has become a serious nosocomial pathogen due to its persistence in the hospital environment and its broad antimicrobial resistance patterns. This review summarizes the most recent literature pertaining to the clinical management of infections with this bacteria emphasizing in-vitro antimicrobial resistance patterns and antimicrobial efficacy in animals and humans. RECENT FINDINGS: Although this pathogen can be associated with an elevated crude mortality, it only contributes to this mortality in a subset of high-risk patients. Determining in-vitro activity of antimicrobial agents can be problematic due to conflicting results sometimes obtained through different testing methods. There is no simple answer as to the most appropriate antimicrobial therapy secondary to lack of adequate studies. Imipenem/cilastatin, amikacin, ampicillin/sulbactam, colistin, rifampin, and tetracyclines are typically active against these bacteria. It is also not clear if combination therapy is more effective than monotherapy. In cases in which A. baumannii-calcoaceticus complex bacteria are resistant to all available agents, we have prolonged infusion times, increased drug dose, and altered route of instillation, such as nebulized therapy for pulmonary infections with mixed results. A primary goal of A. baumannii-calcoaceticus complex management should be to prevent initial colonization and subsequent infection by adequate infection control. SUMMARY: The A. baumannii-calcoaceticus complex continues to play a significant role in our healthcare systems. Prompt and adequate therapy with agents having in-vitro activity is required once it is established that the bacteria represents infection and not colonization. Aggressive infection control policies should be enforced when this pathogen is identified. PMID- 16258324 TI - Epidemiology and mechanisms of glycopeptide resistance in enterococci. AB - PURPOSE OF REVIEW: This review updates epidemiologic trends and our understanding of glycopeptide resistance in enterococci. RECENT FINDINGS: Colonization and infection rates with vancomycin resistant enterococci continue to increase throughout the world while factors contributing to this rise continue to be defined. While no interventions exist to eradicate colonization, infection control procedures are cost effective and decrease the prevalence of vancomycin resistant enterococcal colonization and infection. New molecular methods show great promise in strengthening our ability to detect colonization with these bacteria. Furthermore, our understanding of the origin of vancomycin resistant enterococci continues to grow. Paenibacillus species found in soil have been found to carry homologues of vanA-associated glycopeptide resistance genes found in enterococci. Also, additional evidence supports previous data that VanB associated resistance may have been horizontally transferred from gastrointestinal tract bacteria to enterococci. Finally, glycopeptide resistance has been transferred to methicillin-resistant Staphylococcus aureus in clinical practice on several occasions. SUMMARY: The prevalence of vancomycin resistant enterococci will likely continue to increase. Implementation of infection control strategies, in conjunction with deployment of advanced technologies for detection of vancomycin resistant enterococci, may curb this rise. The emergence of vancomycin resistant S. aureus is of concern. PMID- 16258325 TI - Recent trends in the epidemiology of non-typhoid Salmonella and antimicrobial resistance: the Israeli experience and worldwide review. AB - PURPOSE OF REVIEW: The epidemiology of non-typhoid Salmonella has changed significantly since the turn of the century. Interestingly, non-typhoid Salmonella epidemiology in Israel mirrors some important global trends, and these new trends are reviewed. Recent research that has shed more light on the true toll of non-typhoid Salmonella epidemic and resistance is also summarized. RECENT FINDINGS: After more than three decades of a persistent rise, reports from Israel, the US, and the UK indicate that the trend may be reversed and the incidence of NTS illnesses is starting to decline. In contrast, the rates of resistance and multidrug resistance are increasing and expanding worldwide. Of major concern are the increasing rates of multidrug resistance in Salmonella typhimurium, particularly definitive phage-type 104, the alarming increase in low level ciprofloxacin resistance among several non-typhoid Salmonella serotypes, and the upsurge of high-level ciprofloxacin resistance, mainly in Taiwan. In Israel, high rates of resistance were reported for Salmonella virchow, which accounts for 16% of non-typhoid Salmonella illnesses, and is highly invasive in children. The true burden of Salmonella illnesses in the US was calculated as 520 cases per 100 000, compared with an annual incidence of 13.4 per 100 000 of laboratory confirmed cases. Hospitalization and death rates were 20% and 0.6%, respectively. Infection with resistant non-typhoid Salmonella isolates, and particularly S. typhimurium, increases the likelihood of hospitalization and death. SUMMARY: Many important trends of non-typhoid Salmonella epidemiology are not restricted to a single geographic location, but spread worldwide, reflecting the global nature of the epidemic. This epidemic imposes a heavy burden worldwide. PMID- 16258326 TI - Current concepts in travelers' diarrhea: epidemiology, antimicrobial resistance and treatment. AB - PURPOSE OF REVIEW: The aim of this article is to review recent advances in travelers' diarrhea, which remains one of the most common health problems afflicting individuals from developed countries visiting less affluent regions of the world. RECENT FINDINGS: A large epidemiologic study done at the point of departure provided insights into regional risk factors for travelers' diarrhea and demonstrated that visitors rarely exercised dietary precautions aimed at disease prevention. A preventive program implemented with the close interaction between public health agencies, hotel industry and academia can result in effective reduction of cases. A polymorphism in the IL-8 gene promoter is associated with susceptibility to diarrhea due to enteroaggregative Escherichia coli. New diagnostic tools assist in better understanding the role of norovirus and emerging bacterial enteric pathogens such as enteroaggregative E. coli. Rifaximin, a non-absorbable antibiotic, is a safe and effective alternative for the prevention and treatment of travelers' diarrhea due to non-invasive organisms. SUMMARY: Traditional public health and new antimicrobial agents can decrease the risk of travel related diarrhea. PMID- 16258327 TI - Comparison of the efficacy and pharmacology of formulations of amphotericin B used in treatment of leishmaniasis. AB - PURPOSE OF REVIEW: Several lipid-based formulations of the antifungal and antiparasitic drug amphotericin B are now available on the market. The purpose of this review is to assess their efficacy against leishmaniasis in both experimental and clinical settings, and to point out new developments in the formulation of this antibiotic. RECENT FINDINGS: The development of resistance to pentavalent antimony compounds has shifted the emphasis to amphotericin B for the treatment of visceral leishmaniasis in India. Lipid formulations show good efficacy but are expensive. The treatment period with lipid formulations is shorter, however, which reduces hospitalization costs. As a result, in developed countries where these costs are an important proportion of the treatment, lipid formulations are preferred, whereas they remain largely inaccessible in developing countries. Lipid-associated amphotericin B has been found to be effective for secondary prophylaxis in HIV-positive patients, in studies carried out in European countries bordering the Mediterranean. SUMMARY: The reduced toxicity of lipid-based formulations of amphotericin B is no longer in doubt. In India, their efficacy against visceral leishmaniasis and shorter treatment periods compared with the conventional formulation with deoxycholate has to be counter-balanced against the very high cost. By contrast, in developed countries around the Mediterranean, where leishmaniasis occurs mainly in immunocompromised individuals, lipid formulations have become the treatment of choice for visceral disease. The efficacy against cutaneous lesions is variable, however, and in some reports oral miltefosine was active after failure of treatment with amphotericin B. PMID- 16258329 TI - Stability of transmitted drug-resistant HIV-1 species. AB - PURPOSE OF REVIEW: The proportion of patients showing primary infection with HIV 1 that are already resistant to antiviral drugs has been steadily increasing. Such resistance is known to compromise therapy response. Most patients, however, are not diagnosed during their incident infection and, until recently, it was not known for how long resistant virus can be detected following infection. This article reviews reports on the stability of transmitted resistance in the absence of therapy. RECENT FINDINGS: It has now been shown that many resistance associated mutations can persist for a considerable time following transmission in the absence of treatment and that such resistance can be further transmitted. SUMMARY: Since transmitted resistance can be detected years after infection, it is now worthwhile carrying out resistance tests on newly diagnosed patients in which it is known that the level of transmitted resistance in the population warrants such testing. PMID- 16258328 TI - Artemisinin-based combinations. AB - PURPOSE OF REVIEW: Artemisinin-based combination treatments have been the mainstay of treatment for falciparum malaria in Southeast Asia for more than 10 years and are now increasingly recommended as first-line treatment throughout the rest of the world. RECENT FINDINGS: A large multicentre randomised trial conducted in East Asia has shown a 35% reduction in mortality from severe malaria following treatment with parenteral artesunate compared with quinine. There is increasing evidence that artemisinin-based combination treatments are safe and rapidly effective. Artemether-lumefantrine (six doses) has been shown to be very effective in large trials reported from Uganda and Tanzania. A once daily three dose treatment of dihydroartemisinin piperaquine, a newer fixed combination, was a highly efficacious and well tolerated treatment for multi-drug resistant falciparum malaria in Southeast Asia. SUMMARY: Early diagnosis and treatment of uncomplicated malaria with effective drugs remains a priority as part of a comprehensive malaria control strategy. Artemisinin-based combination treatments have consistently been shown to be highly effective and safe. The challenge is to make them accessible in tropical countries. PMID- 16258330 TI - Host determinants of antiretroviral drug activity. AB - PURPOSE OF REVIEW: As physicians are confronted with a diversity of adverse events and variability in response to medication among patients, this review will focus on the available evidence regarding the impact of genetic variation on drug response. Specifically, variation in drug metabolizing enzymes such as the cytochrome P450s, drug transporters and human leucocyte antigen (for idiosyncratic drug reactions). The potential role of pharmacogenetics in the management of HIV-1 infected individuals and drug discovery will be discussed. RECENT FINDINGS: Wide inter-individual variability of antiretroviral therapy efficacy and toxicity in HIV-infected patients has been extensively reported in the literature. Since treatment involves multiple drugs and drug classes with the potential for significant variability in drug-host as well as drug-drug interactions, antiretroviral drug therapy represents a significant challenge. Despite this inherent complexity, considerable recent advances have led to a greater understanding of interactions between genetic and host factors that influence the efficacy and toxicity of these agents. SUMMARY: The goal of pharmacogenetics in antiretroviral therapy is to outline differences within a given population that influence drug efficacy and toxicity. Although to date, despite the numerous studies available in the literature, conclusions on how the analysis of the relationship between single nucleotide polymorphisms and drug efficacy may not always be clinically useful. Further studies are warranted to clarify the role of pharmacogenetics and antiretroviral drug management. PMID- 16258332 TI - Bibliography. Current world literature. Antimicrobial agents: bacterial/fungal. PMID- 16258331 TI - New paradigms in the management of HIV and hepatitis C virus coinfection. AB - PURPOSE OF REVIEW: Chronic hepatitis C virus infection is currently one of the leading causes of morbidity and mortality in HIV-infected individuals, mainly in hemophiliacs and intravenous drug users. The bidirectional interferences between hepatitis C virus and HIV have clinical consequences and complicate the management of coinfected individuals. RECENT FINDINGS: There is an increased rate of liver complications among coinfected patients due to the decrease in opportunistic infections resulting from the use of potent antiretroviral therapy and accelerated progression to liver cirrhosis in the HIV setting. Conversely, the risk of hepatotoxicity of antiretrovirals is higher in the presence of chronic hepatitis C. While the standard therapy for hepatitis C in HIV is the combination of pegylated interferon plus ribavirin, overall treatment responses are lower in HIV-coinfected than in hepatitis C virus-monoinfected patients. Moreover, interactions between ribavirin and HIV drugs (i.e. didanosine, zidovudine) are associated with higher risks of side effects. SUMMARY: Given the accelerated progression to end-stage liver disease in coinfected patients, treatment of hepatitis C should be a priority. While hepatitis C therapy should not be denied in the absence of contraindication, it should be re-assessed at week 12 and therapy continued only in patients showing more than 2 log drops in viremia, to avoid side effects. Most recent data suggest that adequate selection of candidates, expert management of side effects, and prescription of appropriate ribavirin doses (in genotypes 1-4) and extending treatment (in genotypes 2-3) all might allow response rates in coinfected patients to approach those seen in hepatitis C virus-monoinfected individuals. PMID- 16258334 TI - Faking it? Simulation in the training of obstetricians and gynaecologists. AB - PURPOSE OF REVIEW: This paper discusses the use of simulation as a training tool in obstetrics and gynaecology. RECENT FINDINGS: Modern medical training and patient pressure for treatment by more experienced clinicians have contributed to a reduction in the training opportunities available to junior doctors. Advances in information technology have led to the successful introduction of simulator based training in many safety-critical industries such as aviation and nuclear power. In this editorial we describe simulation devices that are available to obstetrics and gynaecology and explain how simulation can benefit the patient, trainee and educator. We also explore how simulation could be integrated into obstetrics and gynaecology training programmes. SUMMARY: At present simulation is very much underused as a training tool in medicine, and features little in the postgraduate training curriculum. In obstetrics and gynaecology simulation could be used as an educational tool to assist in (1) transfer of knowledge, (2) practising diagnostic and simple practical skills, (3) surgical skills training, (4) emergency drill training and (5) human factors and team training. Whereas simulation should not be perceived as a replacement for training with real patients, educators should embrace the opportunities that simulation provides and integrate it into current training programmes to maximize training opportunities and patient safety. PMID- 16258335 TI - Recent advances in uterine fibroid embolization. AB - PURPOSE OF REVIEW: To summarize the literature on uterine embolization for fibroids published in 2004 and 2005. RECENT FINDINGS: During the last two years, our understanding of the outcome of uterine fibroid embolization has increased. The outcomes are comparable to those that occur after hysterectomy. Health related quality-of-life studies have confirmed the positive impact of the procedure. Improvement in menorrhagia has been quantified using the alkaline hematin method, objectively confirming the outcome. Recovery is also better understood and quantified, with most patients experiencing only moderate pain over the first few days after embolization. In two pregnancy-outcome studies, an increased frequency of cesarean section occurred and possibly a greater likelihood of abnormal placentation, although the data are too few to draw conclusions at this time. Contrast-enhanced magnetic resonance imaging (MRI) has emerged as the primary tool for assessing the potential of complications following the procedure, and our understanding of vaginal discharge and uterine infarction has been increased as a result of its use. SUMMARY: Within the last few years, uterine embolization has become an accepted therapy for uterine fibroids. The increase in understanding gained in recent years has helped to confirm the effectiveness and relative tolerability of this therapy. PMID- 16258336 TI - Pregnancies of unknown location: diagnostic dilemmas and management. AB - PURPOSE OF REVIEW: This review discusses various aspects of the management of women with pregnancies of unknown location. RECENT FINDINGS: The prevalence of pregnancies of unknown location is dependent on the quality of scanning for a given early-pregnancy unit. The higher the quality of scanning, the better the detection of ectopic pregnancy using ultrasound as a single diagnostic test, which in turn results in fewer women being classified with a pregnancy of unknown location. Varying the discriminatory zone does not significantly improve the detection of ectopic pregnancies in a pregnancy of unknown location population. A single serum human chorionic gonadotrophin, when used in a specialized transvaginal scanning unit, is not only potentially falsely reassuring but also unhelpful in excluding the presence of an ectopic pregnancy. A single-visit approach has also been shown to be ineffective. The vast majority of women with a pregnancy of unknown location are at low-risk for ectopic pregnancy. Traditional strategies are capable of detecting the failing pregnancies of unknown location and intra-uterine pregnancies within a pregnancy of unknown location population, but they lack sensitivity for detecting ectopic pregnancies. This justifies the recent development and use of mathematical modelling techniques to predict ectopic pregnancies in the pregnancies of unknown location population. SUMMARY: New mathematical models have been developed to predict the outcome of pregnancies of unknown location; however, prospective studies are needed to assess the reproducibility of these models in different centres on different populations. Hopefully such models will enable the clinician to correctly classify pregnancies of unknown location earlier, in turn reducing the number of follow-up visits. PMID- 16258337 TI - The short and funneling cervix: when to use cerclage? AB - PURPOSE OF REVIEW: The diagnosis of cervical incompetence remains extremely difficult because there is no diagnostic test available prior to, during or after pregnancy. This review will summarize the latest publications on the use of transvaginal ultrasonography to identify women at high risk of preterm delivery and the use of cervical cerclage in these women. RECENT FINDINGS: Cervical length is not only inversely related to the risk of preterm delivery but also inversely related to the risk of intrauterine infection in women with preterm labor. Furthermore, previous history of preterm delivery is related to the risk of preterm delivery. Cerclage trials on women with short cervical length present conflicting results both in low and high-risk populations. Assessment of risk factors and obstetric history remain important in the diagnosis of cervical incompetence. Women at high risk of preterm delivery due to cervical incompetence should be followed-up with transvaginal measurements of cervical length. Only a minority of these women will develop a short cervical length and will consequently be at high risk of preterm delivery. SUMMARY: A combination of assessment of risk factors, obstetric history and follow-up of cervical length enables us to identify women who benefit from a cervical cerclage. PMID- 16258338 TI - Should we deliver twins electively at 37 weeks' gestation? AB - PURPOSE OF REVIEW: The purpose of this review is to assess the available literature to answer the question, should we deliver twins electively at 37 weeks' gestation? RECENT FINDINGS: There are retrospective studies indicating that the lowest risk of perinatal morbidity and mortality occurs in twins at 37 weeks' gestation. A single, small, randomized trial has been identified comparing elective birth at 37 weeks' gestation with continued expectant management. However, there is currently insufficient information available from randomized controlled trials to indicate whether the benefits of elective birth outweigh the potential harms for both women and their infants. SUMMARY: Whereas elective birth at 37 weeks' gestation may be a safe and effective way to reduce the perinatal mortality and morbidity in twins, further information is required from randomized controlled trials to answer the question of whether we should deliver twins electively at 37 weeks' gestation. PMID- 16258339 TI - Uterine artery Doppler screening for adverse pregnancy outcome. AB - PURPOSE OF REVIEW: This article reviews publications in the area of uterine artery Doppler screening that were published during the past year. RECENT FINDINGS: Studies have shown that assessment of a woman's level of risk for pre eclampsia by uterine artery Doppler performs better than maternal history alone. Combination of maternal history and uterine artery Doppler findings leads to even more accurate assessment of risk, and allows calculation of patient-specific risk. Combining screening with uterine artery Doppler and maternal serum biochemical markers has been explored in some studies, but data remain limited and larger prospective longitudinal studies are needed to asses the effectiveness of these techniques. A number of promising markers for development of the disease have been identified, however, including pregnancy associated plasma protein-A, inhibin-A, activin-A and fibronectin, as well as placental growth factor, vascular endothelial growth factor and soluble fms-like tyrosine kinase 1. SUMMARY: Uterine artery Doppler has proved useful in identifying women at high and low risk for developing complications of uteroplacental insufficiency and may aid in stratifying antenatal care. In addition, it can identify women in whom biochemical markers should be measured in order to develop screening tests and aid in evaluation of the pathophysiology of impaired placentation and pre eclampsia. Whether pharmacological intervention in women identified to be at high risk for pre-eclampsia on the basis of Doppler is effective in reducing the incidence of the disease remains to be established. PMID- 16258340 TI - Recurrent miscarriage: pathophysiology and outcome. AB - PURPOSE OF REVIEW: This article reviews new concepts in the aetiology of recurrent miscarriage, presents new outcome data and evaluates new modalities of treatment for unexplained recurrent miscarriage. RECENT FINDINGS: Preimplantation genetic diagnosis has been considered an option for couples who have structural chromosomal abnormalities or unexplained recurrent miscarriage. The association between thrombophilias and adverse pregnancy outcome is further reviewed. In relation to this, there is increasing support for the use of thromboprophylaxis in improving pregnancy outcome in women with inherited thrombophilias. Nonrandomized studies have shown that the reduction in insulin levels with metformin in insulin-resistant individuals may reduce miscarriage risk by restoring normal haemostasis and improving the endometrial milieu. With respect to immunological concepts there is now evidence to suggest that, in addition to a suppression of maternal cell-mediated immunity, some elements of the innate immune system are activated in successful pregnancies. SUMMARY: With the exception of aspirin and heparin for the prevention of recurrent miscarriage in women with the antiphospholipid syndrome, no other suggested therapies for this heterogeneous group of patients have been evaluated in randomized controlled trials. These include thromboprophylaxis for inherited thrombophilias and use of insulin sensitizers in women with insulin resistance and/or polycystic ovarian syndrome. The role of the innate immune system in pregnancy was recently highlighted, and use of nonspecific therapies to suppress the maternal immune response to pregnancy should be reassessed. PMID- 16258341 TI - Progesterone and pregnancy. AB - PURPOSE OF REVIEW: Progesterone is an essential hormone in the process of reproduction. Although the pharmacokinetics and pharmacodynamics of progesterone have been well studied, its use in the pathophysiology of pregnancy remains controversial. One of these concerns is the way in which the hormone is administered. RECENT FINDINGS: In obstetrics the most frequent uses of progesterone are in the treatment of threatened abortion, prevention of recurrent miscarriage, or in the support of the luteal phase in assisted reproduction programmes, and in threatened preterm labour. Randomized, controlled trials showed that women who received progesterone were statistically significantly less likely to have recurrent miscarriages before 34 weeks, to have an infant with birth weight of 2.5 kg or lower, or to have an infant diagnosed with intraventricular haemorrhage. There is currently, however, insufficient information to allow recommendations regarding the optimal dose, route and timing of administration of progesterone supplementation. SUMMARY: Progesterone has shown to be efficacious when continuation of pregnancy is hampered by immunological factors, luteinic and neuroendocrine deficiencies and myometrial hypercontractility. This may explain the reduction in the incidence of preterm birth in high-risk pregnant women using high-dosage prophylactic progesterone. PMID- 16258342 TI - Informed consent: can a patient ever be fully informed? AB - PURPOSE OF REVIEW: The National Health Service Litigation Authority has issued a warning about the process of asking a patient for their consent prior to a medical procedure. This warning was issued in the light of the case of Chester v. Afshar. For the first time in English law the courts have appeared to state that failure to give a patient adequate information about a procedure is negligent per se. This article briefly examines the history of consent since the famous case of Bolam and reviews the recent legal commentary on the case of Chester. It will also consider a proposed solution to the question 'What is adequate information?' RECENT FINDINGS: The medicolegal literature traces the change in the legal test used to determine whether a patient has been adequately informed. It charts the evolution of a 'prudent patient' test and suggests ways in which medical practitioners might adequately fulfil their duty to inform patients properly. SUMMARY: Since the case of Chester v. Afshar it has become harder for a doctor to escape a charge of negligence if they have given inadequate information at the time of asking a patient for their consent to undergo a medical procedure. It is in everyone's interests - doctor and patient - to make the process of consent transparent and to an agreed national standard. PMID- 16258344 TI - Diagnostic and therapeutic capabilities of ultrasound in the management of pelvic pain. AB - PURPOSE OF REVIEW: This review discusses the current diagnostic and therapeutic role of ultrasound in the management of pelvic pain. RECENT FINDINGS: Recent advances in ultrasound technology and expertise have facilitated the accurate diagnosis of common gynaecological and nongynaecological pathologies. Peritoneal and deep infiltrating endometriosis can now be diagnosed using hard and soft ultrasound-based markers. The combination of ultrasound-guided aspiration and instillation of a sclerosant is an alternative to surgery in the management of adnexal masses. SUMMARY: Experience is a key factor in the ability of transvaginal ultrasound to characterize common gynaecological disorders with accuracy. Therapeutic ultrasound provides an alternative to surgery. PMID- 16258343 TI - Short-term and long-term effects of obstetric anal sphincter injury and their management. AB - PURPOSE OF REVIEW: During the past decade increasing attention has focused on the problem of obstetric anal sphincter damage. Although risk factors are now well known, the effects of such damage have received less study. This review focuses on the early and long-term problems that may arise subsequent to anal sphincter injury following childbirth and assesses therapeutic options. RECENT FINDINGS: Up to 25% of women experience altered faecal continence after vaginal delivery, with 4% having persistent symptoms. In those women who have sustained a recognized tear to the sphincter, the quality of primary repair is crucial. Nevertheless, evidence clearly supporting the superiority of overlap over approximation repair is still lacking. The importance of pudendal nerve damage in the aetiology of postpartum faecal incontinence is gaining increasing attention. Augmented biofeedback physiotherapy is the gold standard for treatment of women with such injury, whereas sacral nerve stimulation represents a newer treatment option. SUMMARY: The short-term and long-term effects of obstetric anal sphincter injury warrant increased attention, because with increasing longevity more women are surviving into their 80s and the prevalence of faecal incontinence in this population will increase if measures are not taken to address the problem. Prevention of such injury is not always possible and management options must be further explored. Adequate primary treatment of third-degree tears is of paramount importance. PMID- 16258346 TI - Bibliography. Current world literature. Women's health. PMID- 16258345 TI - Twin-to-twin transfusion: update on management options and outcomes. AB - PURPOSE OF REVIEW: The diagnosis and treatment of twin-to-twin transfusion syndrome has progressed to a staging system to allow directed therapy with the addition of laser to traditional serial amnioreduction. The management options and outcomes are reviewed here. RECENT FINDINGS: In three observational and one randomized controlled trial, laser photocoagulation of chorionic plate vessels at the intertwin membrane improved perinatal survival of at least one fetus and reduced neurological morbidity. Cerebral palsy continues to be a major contributor to adverse outcome with rates of around 20% for survivors. SUMMARY: Treatment strategies for this condition have remained controversial, but two main approaches have been commonly used. Serial, aggressive amnioreduction and fetoscopic laser photocoagulation of the chorionic plate vascular anastomoses at the intertwin membrane. Using the former technique, survival rates of between 18 and 83% have been described. However, 5-58% neurological morbidity has been demonstrated in the surviving infants treated by serial amnioreduction alone. Laser photocoagulation has been advocated in a few specialist centres. Such treatment has been associated with survival rates of between 55 and 69% and potentially reduced neurological morbidity of between 5 and 11% in surviving infants. PMID- 16258347 TI - ARDS diagnosis and management: implications for the critical care nurse. AB - Acute Respiratory Distress Syndrome (ARDS), an intense form of hypoxemic respiratory failure, may be one of the most elusive diagnoses encountered in the intensive care unit. Increasing the knowledge base of the critical care nurse is imperative to prevent and diagnose ARDS, as well as to generate and implement evidence-based clinical interventions. This article presents a thorough examination of the many facets of ARDS, including its definition, etiology, pathophysiology, presentation, diagnosis, and management. PMID- 16258350 TI - The role of gastric pH testing with small-bore feeding tubes: in the intensive care unit. AB - In this observational study, we sought to assess the usefulness of pH values from residuals in intensive care unit (ICU) patients with small-bore feeding tubes. A review of the literature demonstrated that most critically ill patients were excluded in previous studies on gastric pH testing, suggesting that pH testing in critical care is not warranted. Our results demonstrated that 21% of intestinal testing and 64% of gastric testing yielded potential misinformation to the bedside nurse with regards to possible tube location. In conclusion, pH testing for small-bore tube placement and maintenance was not deemed meaningful in our critical care unit. PMID- 16258352 TI - End-of-life care: practical tips. AB - End-of-life care is one part of nursing that many nurses are not fond of administering. The procedure for administering end-of-life care and the rationales for providing such care can benefit the nurse, patient, and family. Considering religious beliefs and practices can assist in proper care for the patient. PMID- 16258353 TI - A personal reflection: leadership: many hats. AB - This article describes one nurse's experiences as a leader in many different roles. PMID- 16258355 TI - The road to JCAHO disease-specific care certification: a step-by-step process log. AB - In 2002, the Joint Commission on Accreditation of Healthcare Organizations (JCAHO) implemented Disease-Specific Care (DSC) certification. This is a voluntary program in which organizations have their disease management program evaluated by this regulatory agency. Some of the DSC categories are stroke, heart failure, acute MI, diabetes, and pneumonia. The criteria for any disease management program certification are: compliance with consensus-based national standards, effective use of established clinical practice guidelines to manage and optimize care, and an organized approach to performance measurement and improvement activities. Successful accomplishment of DSC certification defines organizations as Centers of Excellence in management of that particular disease. This article will review general guidelines for DSC certification with an emphasis on Primary Stroke Center certification. PMID- 16258357 TI - Critical care nurses' perceptions of and responses to moral distress. AB - Nurses frequently experience conflict regarding healthcare decisions, yet are expected to implement actions which they perceive to be morally wrong. Research has described the deleterious effects of this moral incongruency, coined moral distress, on nurses' well being and has identified it as a causative agent in nursing turnover, burnout, and nurses leaving the profession. Thus, it is known that moral distress has significant consequences for nurses, but does moral distress affect nurses' provision of care, and if so, how? PMID- 16258358 TI - Development of the implanted devices adjustment scale. AB - Advances in microelectronics have resulted in exponential growth in the number of implanted medical devices. Most people do well adjusting to their devices, but others show signs of depression and/or anxiety. The Implanted Device Adjustment Scale (IDAS) was developed to measure how well a person is adjusting to an implanted device. First, a pool of items was generated and reviewed by 2 panels of clinicians and psychometricians for content validity. The revised version was then administered to a small sample that provided information about problematic items. Finally, a convenience sample of 45 persons (66% males) with implanted devices (18 pacemakers only, 37 cardioverter/defibrillators) completed the revised IDAS twice. After deleting weak items, the Cronbach alpha was 0.90. No age, gender, or device differences were found. Test-retest reliability was 0.92. The IDAS may be useful to evaluate how well a person is adjusting to her/his device. This may lead to more timely and appropriate interventions to improve outcomes. PMID- 16258362 TI - A closing word: eating our young. PMID- 16258363 TI - Recent advances on the molecular mechanisms involved in pancreatic cancer progression and therapies. AB - This review describes the recent advances in the molecular events involved in pancreatic cancer initiation, progression, and metastasis. Additionally, the importance of deregulated cellular signaling elements as potential targets for developing novel therapeutic strategies against incurable forms of pancreatic cancer is reported. The emphasis is on the critical functions gained by numerous growth factors and their receptors, such as epidermal growth factor receptor, hedgehog signaling, and proangiogenic agents such as vascular endothelial factor and interleukin-8 for the sustained growth, survival, and metastasis of pancreatic cancer cells. The molecular mechanisms associated with antitumoral properties and the clinical benefits of gemcitabine alone or in combination with other cytotoxic agents for the treatment of pancreatic cancer are discussed. PMID- 16258364 TI - Nonpeptidic small-molecule inhibitor of Bcl-2 and Bcl-XL, (-)-Gossypol, enhances biological effect of genistein against BxPC-3 human pancreatic cancer cell line. AB - OBJECTIVES: In pancreatic cancer, several important survival molecules such as EGFR, NF-kappaB, and Bcl-2 or Bcl-XL are highly activated. Thus, agents that inhibit NF-kappaB activation, together with agents that directly inhibit Bcl-2 or Bcl-XL protein function, may lead to enhanced cell killing. (-)-Gossypol, a natural polyphenolic compound isolated from cottonseeds, is a dual and potent small-molecule inhibitor of Bcl-2 and Bcl-XL proteins, with a Ki value in the 300 600 nM range for both proteins. METHODS: : The BxPC-3 human pancreatic cell line was used in this study. (-)-Gossypol was dissolved in DMSO at 20 mmol/L as stock solution, and genistein was dissolved in 0.1 M Na2CO3 to make a 10 mM stock solution. For cell viability, apoptosis, and NF-kappaB studies, MTT assay, histone/DNA ELISA, and Electrophoretic Mobility Shift Assay (EMSA) were used, respectively. Coimmunoprecipitation experiments were designed to study Bcl-XL/Bim heterodimerization, and Western blots to study cytochrome c release. RESULTS: (-) Gossypol showed a concentration-dependent growth inhibition effect against BxPC-3 pancreatic cancer cell line and induced apoptosis with no effect on normal peripheral blood lymphocytes. Results from coimmunoprecipitation experiments indicate that the effect of (-)-gossypol is mediated, at least, in part via disrupting the heterodimerization of Bcl-XL with Bim in BxPC-3 pancreatic cancer cells. (-)-Gossypol completely disrupts Bcl-XL/Bim heterodimerization with no change in the total Bcl-XL or Bim protein, indicating that (-)-gossypol treatment does not affect the levels of Bcl-XL and Bim proteins. We have previously shown that genistein, a prominent soy isoflavone, transcriptionally down-regulates Bcl 2, Bcl-XL, VEGF, MMP-9, and uPAR via inhibiting NF-kappaB activity. In this study, genistein down-regulated NF-kappaB DNA binding activity and inhibited the growth of BxPC-3 pancreatic cancer cells. In addition, the combination of (-) gossypol with genistein showed significantly greater growth inhibition compared with either agent alone. CONCLUSION: From these results, we conclude that inhibition of NF-kappaB activity and direct inhibition of Bcl-2 or Bcl-XL function should serve as a novel strategy for pancreatic cancer therapy. PMID- 16258365 TI - Advanced qRT-PCR technology allows detection of the cholecystokinin 1 receptor (CCK1R) expression in human pancreas. AB - OBJECTIVES: To help clarify the controversy over the detection of expression of the cholecystokinin 1 receptor (CCK1R; CCKAR) in human pancreas. METHODS: Applied qRT-PCR to detect CCK1R expression using the SYBR green/Smart Cycler II and the QZyme oligonucleotide/ABI PRISM 7500 systems to detect CCK1R expressed message in highly purified cDNAs from human pancreas and other tissues. Samples of normal pancreas were obtained at operation (pancreaticoduodenectomy; Whipple's procedure) and used to ascertain the expression of CCK1R in human tissue and investigate donor individual variability in expression levels by semi quantitative RT-PCR and scanning densitometry. RESULTS: We present molecular evidence obtained with advanced qRT-PCR technology that clearly establishes CCK1R expression in human pancreas. Amplification variation in individual human samples is documented here. By targeting different stretches of the sequence with several primer pairs, it was observed that SYBR green qRT-PCR failed to amplify efficiently over GGA-and GAA-rich nucleotide triplet regions, leading to false negative results. The QZyme system quantified the expression with the following distribution: stomach > small intestine approximately colon > brain approximately kidney > pancreas. CCK1R expression levels varied from undetectable, to high levels of expression, in individual samples collected from surgical specimens. CONCLUSION: CCK1R message can be conclusively detected and quantified in human pancreas cDNA by targeting the appropriate nucleotide sequence regions of this gene. PMID- 16258367 TI - [1,2-13C2]-D-glucose profiles of the serum, liver, pancreas, and DMBA-induced pancreatic tumors of rats. AB - OBJECTIVES: In vitro stable isotope glucose tracer studies indicate that undifferentiated cells of the pancreas use glucose primarily through the nonoxidative reactions of the pentose cycle for nucleic acid ribose synthesis, whereas normal or less transformed cells primarily use the oxidative branch of the cycle. METHODS: The pancreatic heads of 4 groups (5/group) of male rats were implanted with time-release pellets designed to deliver placebo or 7,12 dimethylbenzanthracene (DMBA) at 11, 33, or 56 mg/d. Four weeks after pancreatic exposure to DMBA, [1,2-C2]-D-glucose tracer (1 g/kg) was injected intraperitoneally followed by sera collection at 1 and 2 hours and harvest of tumors, adjacent pancreatic tissue, and sera at 3 hours. RESULTS: Tumors (2-9 mm) were found across DMBA groups, with the largest in the high-dose group (> or =5 mm). Selective monitoring by gas chromatography-mass spectrometry of the doubly labeled [1,2-C2]-D-ribose of RNA, which requires nonoxidative synthesis in the pentose cycle, showed a 2.8-, 2.9-, and 5.7-fold increase in pancreatic tumors. Liver and adjacent pancreas preferentially produced [1-C1]-D-ribose through the oxidative reactions of the cycle. Tumor-bearing animals also cleared and recycled tracer glucose at a faster rate. CONCLUSIONS: Simultaneous selective positional ion monitoring of C-labeled metabolites and their mass isotopomers in tissues and blood opens new avenues for the early detection and response to therapy testing of pancreatic cancer using GC-MS and/or magnetic resonance imaging-based methods. This study emphasizes the benefits of stable isotope-based dynamic metabolic profiling, when applied in vivo, and the several advantages it offers to positron emission tomography. PMID- 16258366 TI - Spontaneous activation of pancreas trypsinogen in heat shock protein 70.1 knock out mice. AB - OBJECTIVES: Heat shock proteins (Hsp's) protect cellular proteins in response to injury, and the role of Hsp70 in experimental pancreatitis was recently described. To find out the possible role of Hsp70 in pancreatitis, we used Hsp70 knock-out mice (Hsp70.1-/-) and wild-type mice (Hsp70.1+/+). METHODS: We studied enzymes activities, Hsp70 protein levels, and histologies in cerulein-induced pancreatitis of Hsp70.1-/- and Hsp70.1+/+ mice. RESULTS: In the basal state, Hsp70 protein levels were higher in Hsp70.1+/+ than in Hsp70.1-/- mice, and trypsin activity was higher in Hsp70.1-/- than in Hsp70.1+/+ mice. The zymogen/lysosome ratio of cathepsin B activity before cerulein injection was higher in Hsp70.1-/- than in Hsp70.1+/+ mice. The expression level of Hsp70 in the pancreas increased in both of Hsp70.1-/- and Hsp70.1+/+ mice after hyperthermia because of the Hsp70.3 gene left intact in Hsp70.1-/- mice. After cerulein hyperstimulation, trypsin activity increased 2-fold in Hsp70.1+/+ mice, but cerulein did not further increase basally elevated trypsin activity in Hsp70.1-/- mice. Hyperthermia pretreatment not only blocked cerulein-induced trypsinogen activation, pancreatic edema, and vacuolization in Hsp70.1+/+ mice, but also decreased basally elevated trypsin activity in Hsp70.1-/- mice. CONCLUSIONS: Hsp70 can be responsible for inhibition of cerulein-induced pancreatitis and prevention of spontaneous trypsinogen activation in mice by inhibiting the colocalization of zymogen and lysosomal enzymes. PMID- 16258368 TI - Histopathological diagnosis of pancreatic intraepithelial neoplasia and intraductal papillary-mucinous neoplasms: interobserver agreement. AB - OBJECTIVES: The goal of this study was to evaluate the consistency of distinction between pancreatic intraepithelial neoplasia (PanIN) and intraductal papillary mucinous neoplasms (IPMN) and the hypothesis that guidelines for their distinction might be inadequate. METHODS: A group of 93 pancreas specimens from surgical resections or autopsies that contained lesions consistent with histopathological diagnoses of PanIN-1A, PanIN-1B, PanIN-2, or IPMN (adenoma or borderline) was collected. The classification of these neoplasms by 6 pathologists, 2 from Europe, 2 from Japan, and 2 from the United States, was compared. The pathologists initially used guidelines current in their practice and then reviewed 47 of the 93 specimens a second time using new consensus definitions and guidelines for PanIN and IPMN that were developed in 2003. RESULTS: The initial comparison showed frequent disagreement regarding both category and grade of the lesions. Agreement was greater for category than grade. In the second review, agreement among the 6 reviewers improved, remaining higher for category, although disagreements persisted for both category and grade. CONCLUSIONS: We conclude that the new definitions of PanIN and IPMN improve the consistency in classifying these lesions, but additional work is needed to further improve the reproducibility of their classification. PMID- 16258369 TI - Mutations of the CFTR gene in idiopathic pancreatic hyperenzymemia. AB - OBJECTIVES: Idiopathic pancreatic hyperenzymemia is a new syndrome that is characterized by a chronic increase of serum pancreatic enzymes in the absence of pancreatic disease. The aim of this study was to assess whether mutations of the cystic fibrosis transmembrane conductance regulator (CFTR) gene may have a role in the etiology of this hyperenzymemia. METHODS: Seventy subjects with idiopathic pancreatic hyperenzymemia, 44 men and 26 women (mean age, 48 years; range, 8-74 years), were studied. Thirteen of these 70 subjects had the familial form of the syndrome. The mutation analysis of the CFTR gene was carried out using diagnostic commercial kits for the simultaneous detection of 29 mutations and Tn polymorphism. RESULTS: Among the 70 subjects studied, 7 (10.0%) had CFTR gene mutations. None of these 7 subjects had the familial form of pancreatic hyperenzymemia. These mutations were DeltaF 508 in 1 subject, 2789 + 5 G > A in another subject, and T5 allele in the remaining 5. All these mutations were heterozygous, with the exception of 1 T5 allele that was homozygous in 1 subject. CONCLUSIONS: The frequencies of the mutations of the CFTR gene found in these subjects are similar to the carrier frequencies in the general Italian population. This finding does not support a role for CFTR gene mutations in the etiology of idiopathic pancreatic hyperenzymemia. PMID- 16258370 TI - Enteral feeding without pancreatic stimulation. AB - OBJECTIVE: All forms of commonly practiced enteral feeding techniques stimulate pancreatic secretion, and only intravenous feeding avoids it. In this study, we explored the possibility of more distal enteral infusions of tube feeds to see whether activation of the ileal brake mechanism can result in enteral feeding without pancreatic stimulation, with particular reference to trypsin, because the avoidance of trypsin stimulation may optimize enteral feeding in acute pancreatitis. METHODS: The pancreatic secretory responses to feeding were studied in 36 healthy volunteers by standard double-lumen duodenal perfusion/aspiration techniques over 6 hours. Subjects were assigned to no feeding (n = 7), duodenal feeding with a polymeric diet (n = 7) or low-fat elemental diet (n = 6), mid distal jejunal feeding (n = 11), or intravenous feeding (n = 5). All diets provided 40 kcal/kg ideal body weight/d and 1.5 g protein/kg ideal body weight/d. Plasma gut peptide responses were monitored in 15 subjects. RESULTS: In comparison with basal fasting trypsin secretion rates (mean = 134 [standard error = 22] U/h), duodenal feeding with the polymeric and elemental formulae stimulated trypsin secretion (mean = 408 [standard error = 51] U/h; P < 0.001), whereas intravenous feeding (mean = 171 [standard error = 34] U/h) and mid-distal jejunal (mean = 119 [standard error = 16] U/h) did not. Stimulation was associated with an increase in plasma cholecystokinin, whereas distal jejunal feeding resulted in an increase in plasma glucagon-like peptide-1 and peptide YY concentrations. CONCLUSIONS: Our results suggest that enteral feeding can be given without stimulating pancreatic trypsin secretion provided it is delivered into the mid distal jejunum. The mechanism may involve activation of the ileal brake mechanism. PMID- 16258371 TI - Delayed neutrophil apoptosis attenuated by melatonin in human acute pancreatitis. AB - OBJECTIVES: This study evaluated the expression of neutrophil apoptosis and the effects of melatonin at different concentrations on delayed neutrophil apoptosis in different severities of acute pancreatitis in patients. METHODS: The study population was comprised of 10 patients with severe acute pancreatitis (SAP) and 10 with mild acute pancreatitis (MAP). A total of 10 mL of blood was drawn 24 hours after the onset of the clinical disease for isolation and incubation of the human neutrophils with 4 different concentrations of melatonin. Neutrophil apoptosis activity, CD18 expression, and respiratory burst activity were assessed with flow cytometry 12 hours after incubation. Another group of neutrophils from a healthy control group was used (n = 6) for comparison. RESULTS: Neutrophil apoptosis in patients with SAP is delayed compared with that of patients with MAP. Neutrophils from patients with SAP or MAP are functionally activated. Melatonin at concentrations of 10(-8), 10(-7), or 10(-6) M reverses the delayed process and enhances apoptosis activity in neutrophils in patients with MAP. Melatonin at concentrations of 10(-7) and 10(-6) M reverses the delayed process and increases apoptosis activity in neutrophils in patients with SAP. Neutrophils from patients with SAP and MAP showed significantly increased CD18 expression and respiratory burst activity. Melatonin at concentrations of 10(-7) or 10(-6) M reverses CD18 expression and respiratory burst activity in neutrophils in patients with SAP. CONCLUSIONS: This study highlights the importance of neutrophil apoptosis in patients with SAP and raises the possibility of a therapeutic strategy. Study data show that melatonin promotes neutrophil apoptosis in human acute pancreatitis. PMID- 16258372 TI - Persistent destruction of the basement membrane of the pancreatic duct contributes to progressive acinar atrophy in rats with experimentally induced pancreatitis. AB - BACKGROUND AND AIMS: The imbalance between synthesis and degradation of extracellular matrix (ECM) proteins is a characteristic feature in chronic pancreatitis. We evaluated the relationship between type IV collagen structure in the basement membrane (BM) and the development of acinar atrophy or the regeneration from acinar injury. METHODS: Three different models of pancreatitis were induced in rats by repetitive intraperitoneal injections of 500 mg/100 g body weight of arginine (Arg) or 20 microg/kg body weight of caerulein (Cn) or a single retrograde intraductal infusion of 40 microL/100 g body weight of 3% sodium taurocholate (NaTc). We examined the changes in type IV collagen structure by immunostaining, and the serial changes in the gelatinolytic activity of pro- and active matrix metalloproteinase-2 by zymography in these models of pancreatitis. RESULTS: The pancreas appeared to be histologically normal on day 35 after the first intraperitoneal Cn injection and on day 42 after intraductal infusion of NaTc, whereas 85% to 90% of acinar tissue was replaced by fatty tissue and dilated pancreatic ducts on day 54 after the first intraperitoneal Arg injection. Immunoreactivity for type IV collagen appeared as a discontinuous line along the BM of ducts, vessels, tubular complexes, and acinar cells on day 40 in Arg-induced pancreatitis, whereas it was detected as a continuous line along the BM on day 35 in Cn-induced pancreatitis and on day 42 in NaTc-induced pancreatitits. Gelatinolytic activity of active MMP-2 increased significantly from day 13 to day 40 after the first intraperitoneal Arg injection, whereas it decreased to the baseline level on day 35 after the first intraperitoneal Cn injection and on day 42 after intraductal infusion of NaTc. CONCLUSIONS: Our findings indicate that a long-term increase in gelatinolytic activity of active MMP-2 in Arg-induced pancreatitis causes continuous disorganization of type IV collagen in the BM and progressive acinar atrophy, whereas a transient increase in gelatinolytic activity of active MMP-2 is involved in the regeneration of type IV collagen structure in the BM and recovery from acinar injury. PMID- 16258373 TI - Gene silencing in the endocrine pancreas mediated by short-interfering RNA. AB - OBJECTIVES: RNA interference as mediated by short-interfering RNA (siRNA) offers a nonviral means to silence genes in tissue; however, few data exist about gene therapy using siRNA in pancreas tissue. To determine if siRNA treatment could silence an endogenous gene in pancreatic islets, we developed a murine model using the endocrine pancreas. METHODS: The insulin 2 (Ins2) gene was targeted with siRNA, and quantitative RT-PCR, fluorescent microscopy, and FACS were used to measure transcript levels and siRNA cellular uptake and transfection efficiency. Isolated pancreatic islets were transfected with siRNA in vitro using a liposomal delivery method in a dose titration (50-400 nM) or pooled from BALB/c mice having received siRNA (100 microg) via hydrodynamic tail vein injection. RESULTS: The Ins2 transcript level was significantly reduced by 55% in vitro with FACS data showing a transfection efficiency over 45% with the 400 nM concentration. In vivo delivery of siRNA to pancreatic islets revealed a 33% reduction in Ins2 mRNA levels, although siRNA was able to be detected in 19% of isolated islet cells. CONCLUSION: We have successfully used RNA interference to silence an endogenous tissue-specific gene (Ins2) in pancreatic islets when transfected in vitro or administered in vivo. PMID- 16258374 TI - In vitro interactions between insulin-producing beta cells and embryonic dorsal root ganglia. AB - OBJECTIVES: It has been suggested that the autoimmune assault in type 1 diabetes is not restricted to islet beta cells but also encompasses intraislet nervous structures. Thus, in addition to modulating islet endocrine function, the nerves may also play a direct pathogenic role in diabetes. A major problem in determining the role of neurons in islet function is to distinguish specific neural effects from those mediated through the vascular system, extrinsic hormones, and/or nutritional factors. The aim of this study was to develop an in vitro system that will enable studies on communication between 1 particular type of neuron and islet cell. METHODS: To achieve this, we cocultured rat islets and rat embryonic dorsal root ganglia (DRG) in a 3-dimensional roller-tube system for 1 to 4 weeks. RESULTS: We found a distinct glucose-induced insulin response throughout the culture period. This response was lower compared with islets cultured alone. DRGs survived better when they were cocultured with islet cells. CONCLUSIONS: The roller-tube coculture system provides a novel in vitro system for exploring the interaction between different subpopulations of neurons and pancreatic beta cells. Coculture with DRG neurons reduced glucose-induced insulin release from beta cells, indicating that sensory nerve activity inhibits the insulin response. PMID- 16258375 TI - NestinnegCD24low/- population from fetal Nestin-EGFP transgenic mice enriches the pancreatic endocrine progenitor cells. AB - OBJECTIVES: To identify whether Nestin-positve cells or Nestin-negative cells in pancreas enrich potential pancreatic stem/progenitor cells. METHODS: We generated transgenic mice carrying enhanced green fluorescent protein (EGFP) under the control of the nestin second-intronic enhancer and subsequently divided their embryonic pancreatic cells into different subpopulations according to the expression of EGFP and CD24 and characterized these subpopulations by in vitro culture. RESULTS: The EGFP expression correlated well with that of endogenous Nestin. Only the NestinCD24 subpopulation was able to proliferate and generate immature islet-like cell clusters in long-term culture. Immature islet-like cell clusters could be induced to differentiate into insulin-, glucagon-, and somatostatin-positive cells. CONCLUSIONS: Pancreatic endocrine stem/progenitor cells are enriched in the NestinCD24 population of embryonic pancreas. PMID- 16258377 TI - Three-dimensional structure of peripheral exocrine gland in rat pancreas: reconstruction using transmission electron microscopic examination of serial sections. AB - OBJECTIVES: The purpose of this study was to elucidate the 3-dimensional structure of the peripheral pancreatic exocrine gland. METHODS: We observed serial sections of rat pancreatic tissue using a transmission electron microscope and traced the intercalated duct lumina, intra-acinar secretory canaliculi, intercalated duct cells or centroacinar cells, and basement membranes of acini onto a transparent sheet. These traced diagrams were reconstructed. RESULTS: The intra-acinar secretory canaliculus had branches but no anastomosis. The intercellular secretory canaliculus was extended from the central lumen through the space between the lateral surfaces of the acinar cells to the acinar base. Furthermore, the cytoplasmic process of each centroacinar cell was extended along the central lumen and connected to an intercalated duct cell; thus, centroacinar cells with the same structure as intercalated duct cells were not isolated from the intercalated duct cells. CONCLUSIONS: In this study, we elucidated the normal 3-dimensional structure of the peripheral pancreatic exocrine gland. To understand the pathogenesis of chronic pancreatitis, in the future we intend to examine the morphologic changes of pancreatic tissue during the onset and advancement of chronic pancreatitis using animal models. PMID- 16258376 TI - Effect of octreotide LAR dose and weight on octreotide blood levels in patients with neuroendocrine tumors. AB - OBJECTIVES: Octreotide long acting repeatable (LAR) is widely used for the control of symptoms of functional neuroendocrine tumors. At doses of 30 mg/mo, up to 40% of patients require subcutaneous octreotide "rescue" and up to 40% of patients are given more than 30 mg of LAR/mo. Octreotide acetate binds to the sst2 receptor with an affinity (Kd) of approximately 1 x 10(-9) mol/L (approximately equal to 1000 pg/mL), but higher (approximately equal to 10,000 pg/mL) concentrations of octreotide are required to completely saturate this receptor. Octreotide blood level measurement may be useful to guide LAR therapy in symptomatic patients or in patients who have tumor growth on traditional LAR doses. We hypothesize that LAR doses of 60 mg/mo will produce blood levels of 10,000 pg/mL or greater. At identical monthly LAR doses, patients with higher weights will require more medication to achieve similar plasma octreotide levels than individuals with lower body weights. METHODS: Trough plasma, serum, urine, and saliva octreotide levels were obtained from 52 patients with carcinoid syndrome receiving 20 (n = 8), 30 (n = 19), or 60 mg LAR/mo (n = 10). Octreotide levels were determined by radioimmunoassay. RESULTS: The mean +/- SD plasma octreotide levels for patients receiving 20, 30, or 60 mg LAR/mo were 2518 +/- 1020, 5241 +/- 3004, and 10,925 +/- 5330 pg/mL, respectively. Patient weight (kilograms) was inversely related to plasma octreotide levels. There was a significant correlation between plasma octreotide levels and octreotide levels measured in urine, saliva, and serum. CONCLUSIONS: Frequent measurement of octreotide levels may be useful to guide octreotide therapy in patients with poorly controlled symptoms or those patients experiencing tumor growth. PMID- 16258378 TI - Asian-Oceanic Pancreatic Association: conception to actuality. AB - The Asian-Oceanic Pancreatic Association (AOPA) was organized on July 22, 2005 with the purpose of promoting communication, scholarship, and scientific collaboration, as well as the exchange of knowledge in both basic and clinical research and education in Asian and Oceanic countries. The society will host biennial scientific meetings during the Asian Pacific Digestive Week (APDW) and members consist of individuals rather than regional societies. Given the diversity of its members, English was selected as the common language of communication to facilitate the exchange of ideas. The first AOPA symposium, which aimed to describe the pancreatology in various regions, was held in conjunction with the 60th Annual Meeting of the Japan Society of Gastroenterological Surgery (JSGS). The second scientific meeting will be held during APDW 2007. PMID- 16258379 TI - Pancreaticoduodenectomy in adults with congenital intestinal rotation disorders. AB - Congenital intestinal malrotation is a developmental anomaly resulting from interruption of the physiological herniation and return to the abdominal cavity of the midgut during the 6th to 10th week of embryological development. Normal vascular and anatomic relationships used as landmarks during pancreaticoduodenectomy (PD) are altered in patients with congenital malrotation. We present 3 cases of PD in adults with congenital intestinal rotation disorders. Three adult patients with congenital rotational disorders required PD. Two of these patients had bilio-pancreatic tumors, and 1 cadaveric donor underwent total pancreatectomy during pancreas allograft procurement. All patients had arterial and venous anomalies around the celiac trunk and mesenteric vessels, respectively. The midgut and hindgut in each case were shifted toward opposite sides of the abdominal cavity. Modifications to the standard approach to PD were made, and outcomes were favorable in each case. Each patient showed anatomic abnormalities with the need for identifying vascular structures through their expected (or projected) course and location before parenchymal division or ligation of any vessel. This approach becomes crucial in cases of vascular anomalies, such as ones occurring in congenital malformations, and can be used in similar situations encountered during pancreaticoduodenectomy. PMID- 16258380 TI - Successful resection of a liver metastasis from AFP-producing pancreatic cancer resulting in long-term survival: a case report and review of literature. AB - The prognosis of patients with alpha-fetoprotein (AFP)-producing pancreatic cancer is generally very poor. We, however, present a case of a 33-year-old man who survived AFP-producing pancreatic cancer and its metachronous liver metastasis. The patient had undergone a curative total pancreatectomy for AFP producing pancreas head cancer invading to the tail, which was postoperatively confirmed by immunohistochemical staining of the resected specimen. At 1 year and 4 months after the operation, the serum AFP level was elevated to 26 ng/mL, and a 2.6 cm-sized mass was found in the lateral segment of the liver. He underwent lateral segmentectomy, and the tumor was histopathologically diagnosed as having metastasized from the AFP-producing pancreatic cancer removed earlier. The patient is doing well 8 years and 5 months after resecting the liver metastasis, with no signs of recurrence. This case suggests that surgery may also be a treatment of choice for patients with liver metastasis from AFP-producing pancreatic cancer, particularly for those with a metachronous and solitary tumor. It should be remembered that some pancreatic cancers produce AFP and that monitoring of postoperative serum AFP levels in such cases will be useful for early detection of recurrence and selection of appropriate surgical intervention. PMID- 16258381 TI - Autoimmune pancreatitis with hepatic inflammatory pseudotumor. AB - We report a case of autoimmune pancreatitis (AIP) with hepatic inflammatory pseudotumor (IP). The patient was clinically diagnosed as having multiple metastatic tumors originated from cholangiocellular carcinoma as well as autoimmune pancreatitis and underwent left lobectomy of the liver. Histological examination showed a diffuse or dense lymphoplasmacytic infiltration with obliterating phlebitis but an absence of neoplastic proliferation both in the liver tumor and in the biopsy specimen of the pancreas. Abundant IgG4-positive plasma cells were seen in the lesions. This is the first case report that shows a simultaneous occurrence of hepatic IP and AIP, suggesting that these lesions appeared on the background of the recently proposed entity of IgG4-related systemic disease. PMID- 16258382 TI - Acute pancreatitis subsequent to voluntary methomyl and dichlorvos intoxication. AB - INTRODUCTION: Acute pancreatitis subsequent to methomyl (Lannate) had not been reported until 2005, when Markides published the first case. In our study, we report for the first time 2 cases of acute pancreatitis complicating voluntary methomyl intoxication and compare them with 5 cases of pancreatitis subsequent to dichlorvos poisoning admitted to our toxicological unit during the same period, between July 2001 and June 2003. CASE REPORTS: Patients included in this study were seriously poisoned because all developed muscarinic and nicotinic syndromes. Deep coma and respiratory failure requiring mechanical ventilation were noted in all methomyl-poisoned patients and in only 3 dichlorvos-poisoned patients. Acute pancreatitis occurred 24 to 72 hours after dosing and was characterized by painless abdominal paralytic ileus and vomiting. Clinical features and laboratory examinations were normalized by the fifth day under medical treatment. Complications such as intrapancreatic fluid collection occurred later between days 10 and 20 in 1 methomyl-poisoned patient who required secondary surgical drainage and in 1 dichlorvos-poisoned patient who was treated conservatively. Outcome was favorable in all cases. CONCLUSION: The developing of acute pancreatitis is a serious adverse effect following insecticide intoxication and is better known with dichlorvos than methomyl. PMID- 16258383 TI - Development of an intraductal papillary-mucinous neoplasm of the pancreas in a patient with familial adenomatous polyposis. PMID- 16258384 TI - Minutes of the Business Meeting of the American Pancreatic Association Thursday, November 4, 2004. PMID- 16258386 TI - Substitution of methionine 35 inhibits apoptotic effects of Abeta(31-35) and Abeta(25-35) fragments of amyloid-beta protein in PC12 cells. AB - BACKGROUND: Amyloid-beta peptide (AbetaP), the central constituent of senile plaques in Alzheimer's disease (AD) brain, has been shown to be toxic to neuronal cells and that this toxicity is responsible for the progressive cognitive decline associated with this neurodegenerative disease. The precise mechanism of AbetaP action remains to be determined; however, it has been reported that the methionine residue at position 35 plays a pivotal role in the toxicity of the peptide. With this in mind, the present study examines the effect of mutating the methionine to norleucine in the fragments (31-35) and (25-35) of AbetaP, which have methionine at the C-terminal, in order to investigate the influence of this residue on Abeta-mediated toxic effects on PC12 cells. MATERIAL/METHODS: The toxic and apoptotic effects (release of Cyt c, caspase activation, and DNA fragmentation) exerted by the Abeta(31-35) and Abeta(25-35) wild-type peptides and Abeta(31-35)Met-->Nle, Abeta(25-35)Met-->Nle peptides where methionine was substituted with norleucine were investigated on PC12 cells. RESULTS: The results obtained shown that both peptides induce neurotoxicity in PC12 cells via an apoptotic cell death pathway, including cytochrome c release, caspase activation, and DNA fragmentation. Furthermore, this study reveals that these events were completely abrogated in cells exposed to Abeta peptides in which methionine 35 was substituted by a norleucine residue. CONCLUSIONS: On the basis of the results obtained in this study, an additional hypothesis involving the amyloid-beta peptide and the role of Met-35 has been proposed to clarify the mechanisms responsible of neurodegeneration in Alzheimer's disease. PMID- 16258387 TI - Morphine modulation of the ubiquitin-proteasome complex is neuroprotective. AB - BACKGROUND: Over the past several decades, there is a growing need for the development of neuroprotective compounds, e.g, those that can prevent neural death. It was proposed that nitric oxide (NO), when induced by morphine, would produce neuroprotection in a human neuroblastoma cell line when tested concomitantly with compounds that produce intracellular oxidative stress and neuroinflammation. MATERIAL/METHODS: NO involvement in intracellular protein degradation controlled by the ubiquitin-proteasome complex was examined. Experiments were performed examining the following: a) neural cell viability and morphology; b) gene specific mRNA levels via semi-quantitative RT-PCR; c) protein levels via Western blotting; d) enzymatic activity via fluorogenic substrate cleaving assays; and lastly, NO release via the Apollo 4000 real-time amperometric detector. RESULTS: Morphine induces the production of NO in human neuroblastoma cells, which can be blocked by naloxone and the cNOS inhibitor L NAME. Rotenone, which induces oxidative stress and increases the expression of the proteasomal catalytic X subunit, causes the cells to die and morphine inhibits this process via NO. Rotenone also increases the activity of the 20S proteasome, whereas morphine alone or in the presence of rotenone caused a decrease in the activity of the 20S proteasome. Morphine decreases the expression of the immunoproteasome catalytic subunit LMP7 in response to inflammatory stimulation, demonstrating that morphine's neuroprotective action does not apply to only oxidative stress. Morphine significantly increases free ubiquitin, suggesting that morphine is inducing neuroprotection by reducing the amount of oxidized proteins targeted for degradation. CONCLUSIONS: Significant neuroprotection on the cellular and molecular levels was demonstrated and serves as a foundation for future work concerning the development of novel ligands for morphine's mu3 opiate receptor in an effort to prevent cellular death associated with neurodegenerative diseases. PMID- 16258388 TI - Tyrosine and tyramine increase endogenous ganglionic morphine and dopamine levels in vitro and in vivo: cyp2d6 and tyrosine hydroxylase modulation demonstrates a dopamine coupling. AB - BACKGROUND: The ability of animals to make morphine has been in question for the last 30 years. Studies have demonstrated that animals do contain morphine precursors and metabolites, as well as the ability to use some morphine precursors to make morphine. MATERIAL/METHODS: The present study uses excised ganglia from the marine invertebrate Mytilus edulis as well as whole animals. Morphine and dopamine levels were determined by high performance liquid chromatography coupled to electrochemical detection and radioimmunoassay. Tissues and whole animals were also exposed to morphine precursors and exposed to the CYP2D6 inhibitor quinidine and the tyrosine hydroxylase inhibitor alpha-methyl para-tyrosine (AMPT). Additionally, via RT-PCR, a cDNA fragment of the CYP2D6 enzyme in the ganglia of M. edulis was identified. RESULTS: Pedal ganglia incubated with either tyramine or tyrosine, or whole animals receiving injections, exhibited a statistically significant concentration- and time dependent increase in their endogenous morphine and dopamine levels (2.51 +/- 0.76 ng/g for tyrosine and 2.39 +/- 0.64 ng/g for tyramine compared to approximately 1.0 ng/g morphine wet weight). Incubation with quinidine and/or AMPT diminished ganglionic morphine and dopamine synthesis at various steps in the synthesis process. We also demonstrated that CYP2D6 mediates the tyramine to dopamine step in this process, as did tyrosine hydroxylase in the step from tyrosine to L-DOPA. Furthermore, via RT-PCR, we identified a cDNA fragment of the CYP2D6 enzyme in the ganglia, which exhibits 94% sequence identity with its human counterpart. Evidence that tyrosine and tyramine were, in part, being converted to dopamine then morphine, and that this process can be inhibited by altering either or both CYP2D6 or tyrosine hydroxylase, is also provided. CONCLUSIONS: It appears that animals have the ability to make morphine. This process also appears to be dynamic in that the inhibition of one pathway allows the other to continue with morphine synthesis. Moreover, dopamine and morphine synthesis were coupled. PMID- 16258389 TI - The effects of topical estriol and vehicle cream on the mechanical and histological properties of normal rat skin. AB - BACKGROUND: The biomechanical and histological effects of topical estriol in normal rat skin are not clearly defined, although several favourable effects of estrogens have been suggested, especially in the postmenopausal period. Our goal was to determine whether estrogens have any such effects on skin histology and biomechanical properties in an experimental rat model. MATERIAL/METHODS: Estriol cream or vehicle cream was topically applied to the dorsum of Sprague Dawley rats of both genders in three separate groups for two weeks, and the results of the estriol and vehicle groups, as well as the non-treated control groups, were compared in terms of mechanical tensometric test results and histopathological evaluation. RESULTS: After the application of topical estriol, histologically, subcutis thinned (p<0.01), the thickness of collagen fibrils increased (p<0.01), and elastic fibrils and dermal connective tissue cells appeared denser in female estriol group rats; mechanically, the elasticity modulus increased in the estriol groups of both sexes (p<0.05), whereas breaking strength, tensile strength and strain decreased in female estriol group rats (p<0.05). CONCLUSIONS: Estrogens can change the mechanical and histological features of normal skin, and these changes are observed differently in female and male rats. Further studies to investigate the collagen structure and its interaction with ground substance, as well as effects of estriol on rat skin without a vehicle cream, are necessary to better understand the effects of estriol on skin mechanics. PMID- 16258390 TI - Morphological assessment of craniovetebral junction development in the prenatal period. AB - BACKGROUND: Because of numerous abnormalities of craniovertebraljunction development observed in the prenatal period, the region has been studied thoroughly in order to elucidate the mechanism of axial differentiation, dependence of vertebral morphology on the development of ossification centers and the mechanisms regulating the dynamics of growth. AIM: Analysis of morphological changes of the craniovertebral junction taking place during the prenatal period by quantitative assessment of some developmental changes such as growth rate, sexual dimorphism, interdependence of the analyzed parameters, taking into consideration the variable geometry of the studied area. MATERIAL/METHODS: he study was carried out on 48 fetuses (22 male, 26 female) of 86-212mm crown-rump length (CRL) range. The applied methods included: anthropometry, precise measurements, as well as trigonometric calculation of angles. RESULTS: All the investigated parameters demonstrated a strong, statistically significant correlation with CRL. No statistical significance of sexual dimorphism and differences between left- and right-sided measurements was demonstrated. The development rate indexes for the height of C1,C2 posterior vertebral arches, unlike those for the transversal dimensions of C1,C2 posterior vertebral arches, decrease after 5 months of gestational age. Little variability of the angles determined by geometrical reconstruction evidences close interdependence in the development of the analyzed craniovertebral junction elements. CONCLUSIONS: Geometrical configuration of the craniovertebraljunction during the fetal period is constant, with growth isometry preserved throughout intrauterine development. The occipital bone and Cl,1C2 cervical vertebrae are closely correlated according to structural and functional dependences. PMID- 16258391 TI - Kinetic evaluation of human cloned coproporphyrinogen oxidase using a ring isomer of the natural substrate. AB - BACKGROUND: The enzyme coproporphyrinogen oxidase (copro'gen oxidase) converts coproporphyrinogen-III (GIII) to protoporphyrinogen-IX via an intermediary monovinyl porphyrinogen. The A ring isomer coproporphyrinogen-IV (C-IV) has previously been shown to be a substrate for copro'gen oxidase derived from avian erythrocytes. In contrast to the authentic substrate (C-III) where only a small amount of the monovinyl intermediate is detected, C-IV gives rise to a monovinyl intermediate that accumulates before being converted to an isomer of protoporphyrinogen-IX. No kinetic studies have been carried out using the purified human copro'gen oxidase to evaluate its ability to process both the authentic substrate as well as analogs. MATERIALS/METHODS: Therefore, purified, cloned human copro'gen oxidase was incubated with C-III or C-IV at 37 degrees C with various substrate concentrations (from 0.005 pM to 3.5 pM). The Km (an indication of molecular recognition) and Kcat (turnover number) values were determined. RESULTS: The Km value for total product formation was about the same with either C-III or C-IV indicating the same molecular recognition. However, the catalytic efficiency (Kcat/Km) of the enzyme for total product formation was not more than two fold higher using C-III relative to C-IV. CONCLUSIONS: Since the Km values are about the same for either substrate and the total Kcat/Km values are within two fold of each other, this could correlate with the increase of severity of porphyrias with monovinyl accumulation. The ability of the increased levels of C-IV to compete with the authentic substrate has important implications for clinical porphyrias. PMID- 16258392 TI - Antioxidant and hepatoprotective effects of Eucalyptus maculata. AB - BACKGROUND: Since phenolic compounds have been reported as effective antioxidants, this study was designed to assess the hepatoprotective and antioxidant activities of the chloroformic extract of the resinous exudate and its phenolic constituents obtained from the stems of Eucalyptus maculata. MATERIAL/METHODS: The chloroformic extract and pure phenolic isolates were evaluated for their antioxidant and hepatoprotective properties in mice and rats based on biochemical changes in serum and tissues as well as pathological changes in the liver and spleen. RESULTS: Acetaminophen (ACP) at a dose of 1 g/kg body weight produced 100% mortality in mice, while pretreatment of animals with the chloroformic extract (125 and 250 mg/kg) protected against the moralities by 66%. Pretreatment of rats with either the chloroformic extract (250 mg/kg) or any of the pure isolates (20 mg/kg) significantly reduced the increase in serum level of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP) produced by ACP (640 mg/kg). Pretreatment of animals with the chloroformic extract or its isolates also protected against ascorbic acid depletion in serum and kidney tissues induced by oral administration of paraquat (PQ) without modifying the serum level of glutathione (GSH) and glycogen content in liver tissue. CONCLUSIONS: The phenolic content of the chloroformic extract and the pure isolates produced an antioxidant activity which may be due to the formation of stable phenoxyl radical in addition to its effect through vitamin C. PMID- 16258393 TI - Psychosocial situation of living donors: moods, complaints, and self-image before and after liver transplantation. AB - BACKGROUND: Living donor liver transplantation (LDLT) has developed into an established therapy for terminal liver diseases. There is, however, little knowledge of the psychosocial situation of living donors. The aim of this study was to investigate the course of donors' moods, physical complaints, and self image in the preoperative decision process and six months after surgery. MATERIAL/METHODS: Fifty-two potential donors were evaluated at the Charite Berlin. On the basis of the clinical interview, seven (13%) potential donors were not recommended for LDLT for psychosocial reasons. Twenty-six donors underwent liver resection and were re-evaluated six months after surgery. Donors' moods were investigated with the Berlin Mood Questionnaire (BMQ) and their physical complaints were assessed by the Giessen Complaint Questionnaire (GCQ). The Narcissism Inventory (NI-90) was used to assess their self-image. RESULTS: Potential donors who were not recommended for surgery showed significantly higher values for negative moods, physical complaints, and 'narcissistic rage". After surgery, donors' self-images were generally stabilized. Nevertheless, five donors (19%) showed high values for "threatened self" before and after transplantation, as well as high values for negative moods and complaints after LDLT. CONCLUSIONS: The resection of the right hepatic lobe holds promise of a good psychosocial outcome, with stable self-image and self-esteem for most donors. A minority of donors exhibited enhanced perception of distress and low self-esteem before and after surgery, which can easily be overlooked in the preoperative evaluation. A psychometric assessment of self-image before transplantation should be added to the clinical interview evaluation of potential living donors. PMID- 16258394 TI - Serum retinol and total carotene concentrations in obese pre-school children. AB - BACKGROUND: Obesity among pre-school children and its repercussions on adult life are growing problems, but there has been little research focusing on its relationship with the deficiency of serum retinol and carotenoids in our region. MATERIAL/METHODS: In a case-control study, a group of 46 preschool children, were matched by sex and age (23 obese and 23 non-obese subjects; average age 5.74 and 5.76 years, respectively). Serum retinol and carotenoid levels were evaluated according to Underwood and Sauberlich. Other aspects evaluated included nutritional status using the weight/height z-score (Obesity ZWH > or = 2), serum levels of triglicerides (TG), total cholesterol (TC) and the VLDL-c, HDL-c, and LDL-c fractions, classified according to Kwiterovich. RESULTS: Serum retinol insufficiency was 18.2% vs. 6.7% (p=0.38) for cases and controls, respectively. Low carotenoid levels were found in the obese in relation to the non-obese (82% vs. 26,6%, p=0,0054 and OR=12,4). No statistically significant difference between the case and control groups was found for TC and cholesterol fractions, TG and retinol. The findings for the tested parameters were as follows: high TC (cases 30.4%, controls 21.7%; p=0.50), LDL-c (cases and controls 34.8%; p=0.50), low HDL c (cases 17.4%, controls 26%; p=0.47), high TG (cases 31.8%, controls 17.4%, p=0.65) and high VLDL-c (cases 21.7% and controls 8.7%; p=0.20). CONCLUSIONS: The association of obesity, hyperlipidemia and low serum level of carotenoids, which are essential to antioxidant protection, may be one of many factors predisposing obese children to a high risk of atherosclerosis later in life. PMID- 16258395 TI - Sweat production during global heating and during isometric exercise in people with diabetes. AB - BACKGROUND: While sweat production in response to heat is impaired in people with diabetes, sweat production has not been examined during isometric exercise. MATERIAL/METHODS: Eight subjects with type 2 diabetes and 9 control subjects exerted a fatiguing isometric contraction of the handgrip muscles at a tension of 40% of the maximum voluntary strength (MVC) after exposure to a 32 deg C environment for 30 min. compared to 10 controls and 10 subjects with diabetes exposed to a 39 deg C environment. RESULTS: Sweat was impaired to all areas of the body during heat exposure in patients with diabetes under both environmental conditions. For example, on the chest, the average sweat rates after exposure to the 32 deg environment was 259.2 +/- 55.2 nanoliters/min in control subjects and 198.3 +/- 46.2 nanoliters/min for subjects with diabetes. Compared to the 32 deg C environment, control subjects increased sweat in all 4 areas proportionally more than subjects with diabetes. Sudomotor rhythm was present in sweat in control subjects at a rate of repetition of 11 and 50 seconds but almost absent in subjects with diabetes. During exercise, sweat rates slowly increased from the beginning to the end of the exercise. But the head of the subjects with diabetes showed hypersweating while the other areas showed diminished sweating compared to control subjects. CONCLUSIONS: Thus some of the impairment in sweating may be due to central mechanisms associated with heat sensitivity or in the hypothalamus and not to the sweat glands themselves. PMID- 16258396 TI - Changes in body composition and insulin sensitivity in severely obese subjects after laparoscopic adjustable silicone gastric banding (LASGB). AB - BACKGROUND: Laparoscopic adjustable silicone gastric banding (LASGB) is a viable therapeutic approach to achieve stable body weight reduction in severe obesity. The aim of this study was to evaluate body composition and metabolic features in morbidly obese patients before and after LASGB. MATERIAL/METHODS: There were 15 severely obese patients (Ob) (M/F: 4/11; mean age: 32.5 +/- 3.8 years) and 16 age and sex-matched healthy lean controls (C) (M/F: 4/12; mean age: 39.5 +/- 2 years). Body mass index (BMI), waist circumference, waist-to-hip ratio, blood pressure, lipid profile, serum glucose and insulin during OGTT, and HOMA-IR were evaluated in all subjects. Body composition and fat distribution were measured using dual energy X-ray absorptiometry (DEXA). Patients were assessed before and six months after LASGB. RESULTS: The obese subjects showed several metabolic alterations. There were also positive correlations between waist, fat mass (FM), and HOMA-IR at baseline. After LASGB, mean BMI fell from 42.2 kg/m2 to 33.2 kg/m2; waist circumference and abdominal FM% decreased significantly. FM% declined, whereas FFM% increased. The ratio of FM loss to FFM loss was 3.7:1. Serum glucose and insulin levels during OGTT diminished slightly after weight loss and triglyceride levels fell dramatically. After LASGB, fasting insulin and HOMA-IR declined. RESULTS: LASGB induced a significant improvement in insulin sensitivity and a redistribution of body composition with a relative increase of FFM. PMID- 16258397 TI - Antenatal care: a comparison of demographic and obstetric characteristics of early and late attenders in the Niger Delta, Nigeria. AB - BACKGROUND: The utilization of modern antenatal care is poor in most parts of Nigeria and studies indicate that the majority of those who present for antenatal care do so late. MATERIAL/METHODS: This was a retrospective study involving 303 consecutively booked pregnant women over a 6-month period (March 1 to August 31, 2004) in a suburban tertiary hospital practice. The objectives were to determine the incidence of late booking for antenatal care and to compare the demographic and obstetric characteristics of women who booked late with those who booked early for antenatal care. The Fisher's exact test, odds ratio, and the 95% confidence interval were used for statistical analysis. RESULTS: Most (79.9%) of the women booked late for antenatal care. It was found that age, parity, level of education, social class, previous fetal loss, and previous obstetric complications did not differ significantly between women who booked early and those who booked late. CONCLUSIONS: The majority of the study population booked late for antenatal care. Socio-demographic and obstetric factors do not appear to be responsible for late utilization of antenatal care in the Niger Delta, Nigeria. Further studies are needed to explore the contribution of cultural factors to late presentation for antenatal care. Community-based health education programs on the importance of early presentation for antenatal care need to be put in place to address the anomaly of late utilization of antenatal care in Nigerian women. PMID- 16258398 TI - Incidence of the del35G/GJB2 mutation in Croatian newborns with hearing impairment. AB - BACKGROUND: The de135G mutation in the GJB2 gene is the most common cause of prelingual deafness. The mutation frequency has so far been estimated either by testing symptomatic children or adults, or by carrier testing of the general population. The purpose of our study was to establish the incidence of the del35G/GJB2 mutation in newborns with hearing impairment--in congenital deafness. MATERIAL/METHODS: Patients were identified through a neonatal screening program (performed on a regular basis in Croatia since 2002). Otoacoustic emission testing was performed on 3275 newborns, and allele-specific PCR was performed on newborns diagnosed with hearing impairment. RESULTS: Hearing impairment was found in 9 newborns, the frequency of congenital hearing impairment being 1/363; the del35G mutation was found in 3 of these 9 newborns. The established incidence of the mutation in the studied population of Croatian newborns with hearing impairment is 1/1091 (95CI: 1/372-1/3205). CONCLUSIONS: This particular approach to patient identification, based on exact clinical examination supplemented with molecular testing, allowed for complete diagnosis and precise estimation of the incidence of the mutation in cases of congenital deafness, which proved to be higher than previously reported in prelingual deafness. This finding has important implications in clinical evaluation and genetic counseling of patients and their families. PMID- 16258399 TI - Clinicopathological features of lupus anticoagulant; experience at AFIP during last seven years. AB - BACKGROUND: The lupus anticoagulant (LA) belongs to family of immunoglobulins, most commonly an IgG isotype. These antibodies have been identified most frequently in association with thromboembolic events, recurrent fetal loss and thrombocytopenia. The aim of the present study was to evaluate the presenting clinical and pathological features in patients LA positive presenting at AFIP Rawalpindi over the period of Jan 1993 to Nov 2000. MATERIAL/METHODS: Retrospective analysis of patients presenting with positive LA was carried out. RESULTS: 1583 suspected cases were screened for LA including 1370 females and 213 males. 1024 cases presented with history of recurrent abortions, 292 with thrombosis, 152 with thrombotic strokes before the age of 45 years, 52 with thrombocytopenia and 63 with miscellaneous disorders. Out of 1024 patients tested for recurrent abortions, 130 (13%) females were positive for lupus anticoagulant. Ten (6.5%) of 152 patients presenting with strokes were found positive. Out of 292 cases presenting with thrombosis 17 (5.9%) were found positive. CONCLUSIONS: The lupus anticoagulant prevalence in Pakistani patients with recurrent fetal loss, stroke and thrombosis is statistically significant. Clinicians should be made aware of association of LA with various diseases. The successful management of these patients depends upon close liaison with obstetricians, physicians and haematologists. PMID- 16258400 TI - C-reactive protein is an indicator for fatal outcomes in first-time stroke patients. AB - BACKGROUND: The causal relation of high sensitivity C-reactive protein (hsCRP) to ischemic cerebrovascular disease remains unclear, though an inflammatory effect has been suggested in many studies. The aim of our study was to evaluate serum hsCRP levels in ischemic stroke patients and in a control group, and to correlate the values with other generally known risk factors. MATERIAL/METHODS: A total of 47 patients with ischemic stroke and 26 control subjects were recruited. The cases were well-matched to controls for age and sex. Peripheral blood samples from stroke patients were obtained between 12-24 hours after the stroke. Serum levels of hsCRP were determined by chemoluminescence assay. RESULTS: The mean serum levels of hsCRP were found to be significantly higher in patients (3.12 +/- 4.4 mg/dL) than controls (0.39 +/- 0.6 mg/dL, p=0.0001). The level of hsCRP was above the risk limit in 39 patients (83.0%) and 7 controls (26.9%). The hsCRP values were not related to the presence of other vascular risk factors, except for cholesterol level. The correlation analysis of hsCRP revealed a linear correlation with death within six months and the presence of hypertension. CONCLUSIONS: Our results suggest that elevated serum hsCRP levels may be an indicator of fatal outcome in first-time stroke patients. PMID- 16258401 TI - The expression of vWF receptor on newborn platelets. AB - BACKGROUND: Platelets contain a complex known as GPIb-IX-V. The receptor for vWF is located in GPIbalpha. The number of functional receptors decreases during thrombin activation of platelets. This study was performed to evaluate the percentage of platelets displaying the GPIbalpha -IX complex on their surface. MATERIAL/METHODS: We studied 51 healthy term newborns (21 females and 30 males). Blood was collected from the umbilical artery and vein immediately after severance of the umbilical cord, using a Diatube TM CTAD Vacutainer System. The GPIbalpha-IX complex was tested with CD42b-PE antibody, using a EPICS XL flow cytometer. RESULTS: Practically no differences were observed in the percentage of platelets demonstrating CD42b in venous blood (18.59%) and arterial blood (19.37%). The percentage of platelets bearing the CD42b molecule is higher in male newborns (20.58%) than female (17.38%). Healthy control adults demonstrated an increased percentage of CD42b on platelets (30.4%) in comparison to newborns (18.98%). CONCLUSIONS: The sex of neonates affects the tested complex: higher values were noted in males. The particular umbilical vessel (vein or artery) has no effect on the percentage of platelets bearing CD42b. PMID- 16258402 TI - Clinical spectrum and critical care management of Posterior Reversible Encephalopathy Syndrome (PRES). AB - BACKGROUND: Posterior Reversible Encephalopathy (PRES) is characterized by acute onset headache, altered mental status, cortical blindness and seizures, with parietal-occipital involvement. We report all cases of PRES diagnosed in our intensive care unit during the last 4 years, and evaluate their outcome in terms of the different medical treatments used. Even if usually reversible, PRES can sometimes result in death or irreversible neurological deficit, such as chronic epilepsy. MATERIAL/METHODS: From January 2001 to January 2005, we identified 8 female patients with PRES. All patients underwent basal and follow-up brain MRI. Patients referred to Epilepsy Center (about 3000 patients) were reviewed in order to identify subjects with a clinical history of PRES. These latter were clinically re-evaluated and underwent video-EEG, MRI study and neuropsychological testing. RESULTS: Of the 8 patients, 5 had hypertensive encephalopathy during pregnancy; 2 had eclampsia during the postpartum period; 1 patient with chronic renal failure developed symptoms after immunosuppressive treatment. In all patients but 1, neurological and radiological abnormalities resolved after appropriate treatment. In addition, we found 2 patients with temporal lobe epilepsy subsequent to a previous PRES. MRI revealed cortical-subcortical malacia in the parietal-occipital regions. CONCLUSIONS: The widespread use of MRI technology has made PRES familiar to many clinicians. Although PRES is reversible when treatment is instituted, delayed diagnosis and therapy can result in chronic neurological sequelae. The relationship between hypertensive encephalopathy and chronic epilepsy needs to be confirmed by longitudinal studies. Normalization of blood pressure and treatment of seizures deserves particular attention. PMID- 16258403 TI - Anaplastic carcinoma of the pancreas with squamous features: report of a case and immunohistochemical study. AB - BACKGROUND: Anaplastic carcinomas of the pancreas are rare aggressive tumors with survival measurable in weeks. Many terms have been applied used to describe these tumors, and anaplastic foci are identified in ductal adenocarcinomas and in ectopic pancreata, but are not the dominant pattern of growth. We herein present our experience with a case of anaplastic carcinoma of the pancreas with squamous features in order that allowed us to delineate the clinicopathologic and immunohistochemical features of this rare entity. CASE REPORT: According to imaging findings, the 77-year-old Japanese man was diagnosed as the malignant pancreatic tumor, and underwent a surgical resection. Histopathologically, anaplastic tumor cells showed focal ductal and squamous features infiltrated into pancreatic parenchyma, extrapancreatic fatty tissue, and stomach. The tumor cells showed strong reactivity for cytokeratin, alpha-SMA, vimentin, NSE, and S-100 protein. Although immunoreactivity against p53 was negative, strong positive immunostaining for proliferating cell nuclear antigen and interleukin-1 receptor type I (IL-1RI) was observed in a the majority of tumor cells, while the alpha6 integrin subunit was predominantly strong expressed in the adenocarcinomatous lesion. The patient's postoperative course was uneventful and he was treated with a chemotherapy consisting of gemcitabine. After discharging from the hospital, he had subsequently been observed as an outpatient and same chemotherapy was followed by weekly. However, the patient suffered from peritonitis carcinomatosa and re-increases of multiple liver metastases, and he died in the fourteenth month after surgery. CONCLUSIONS: Our immunohistochemical studies suggested that the prognosis of the case with anaplastic carcinoma presented here would be poor, due to the strong expression of integrins and IL-1RI. PMID- 16258405 TI - Intellectual property 101. PMID- 16258404 TI - A case of left ventricular pseudoaneurysm with long survival and congestive heart failure as first presentation. Case report and review of the literature. AB - BACKGROUND: Left ventricular pseudoaneurysm is a rare and extremely fatal complication of acute myocardial infarction. It is defined as a rupture of the myocardium contained by epicardial adhesions or the epicardial wall. CASE REPORT: A 76-year-old woman was brought to our department suffering from acute pulmonary edema and left ventricular pseudoaneurysm. This condition was caused by an unrecognized high lateral myocardial infarction, which was diagnosed and treated conventionally. Four years after the diagnosis was made, the patient is in good physical condition. The presentation, imaging findings, and the prognosis of such cases are briefly discussed. We have especially focused on the imaging techniques currently used to confirm the diagnosis and on the fact that although the patient refused to have an operation (most appropriate for the diagnosis), she still lives in good physical condition (NYHA II). CONCLUSIONS: We report a case of post infarction pseudoaneurysm along with review of the literature on the subject. We discuss the role of computed tomography and magnetic resonance imaging to ascertain the diagnosis and the algorithm of therapeutic management. Based on current literature, we believe that surgical resection is the treatment of choice for patients in whom a pseudoaneurysm is detected within three months after myocardial infarction, for patients with other indications for cardiac surgery, and for symptomatic patients with ventricular tachycardia or recurrent embolism related to the pseudoaneurysm. Optimum medical therapy is the only alternative in those high-risk patients who refuse surgical operation. PMID- 16258406 TI - Regulation of various genes in human leukocytes acutely exposed to morphine: expression microarray analysis. PMID- 16258407 TI - Evaluation of culture, tube agglutination, and PCR methods for the diagnosis of brucellosis in humans. AB - BACKGROUND: Brucellosis is prevalent in Saudi Arabia and Brucela melitensis is a leading cause of zoonosis worldwide. Therefore, accurate diagnosis of brucellosis is a key to its treatment and control. MATERIAL/METHODS: Twenty patients presented with symptoms of brucellosis were examined before and after antibiotic treatment for the diagnosis of brucellosis. Sequential blood samples collected monthly from each patient were tested for the diagnosis of brucellosis by serum plate agglutination test (SPA), standard tube agglutination test (STA), culture, and polymerase chain reaction (PCR). RESULTS: While most of the samples were positive by the agglutination tests, only 40% and 70% were positive by culture and PCR, respectively. After the course of antibiotic treatment, the culture rate and PCR results were positive in 10% of the samples. In contrast, anti-brucella antibodies of the treated patients were positive in 20% and 45% by STA and SPA tests, respectively. Furthermore, agglutinating antibodies in the presence of 2 mercaptoethanol were positive in 60% of the enrolled patients and negative in all patients after the antibiotic treatment. CONCLUSIONS: The present study revealed that the expression of anti-brucella antibodies does not correlate with the status of the disease condition. Further, completion of antibiotic therapy hampered the appearance of brucella-specific IgM antibodies, but did not eliminate the appearance of residual IgG antibodies in the treated patients. Therefore, for effective therapy, detection of the Brucella organisms by PCR or culture is an important attribute in the evaluation of the treatment regimen against brucellosis. PMID- 16258408 TI - Effect of inhaled salbutamol on dynamic intrinsic positive end-expiratory pressure in spontaneously breathing patients with stable severe chronic obstructive pulmonary disease. AB - BACKGROUND: The objective was to determine the effect of inhaled salbutamol on PEEPi dyn in spontaneously breathing patients with stable severe chronic obstructive pulmonary disease (COPD). MATERIAL/METHODS: Eleven patients with COPD were studied (mean age: 63 years, average FEV1 0.71 +/- 0.31 L). The patients were evaluated before and after inhaling 400 pg of salbutamol. Spirometry, slow vital capacity, breathing pattern, and PEEPi dyn pre- and post-bronchodilator administration were performed. PEEPi dyn was measured as the difference in esophageal pressure between the onset of inspiratory effort and the point of zero flow. RESULTS: Pre-BD PEEPi dyn fell on average by 56% after inhalation of salbutamol (1.89 +/- 1.24 cm H2O to 0.83 +/- 0.81 cm H2O, p<0.001). This was accompanied by bronchodilation and a reduction of air trapping (FEV1: 0.71 +/- 0.31 L to 0.91 +/- 0.35 L, p<0.001; IC: 1.5 +/- 0.53 L to 1.94 +/- 0.56 L, p<0.001). A significant positive correlation between deltaPEEPi dyn, deltaFEV1 (r=0.64, p<0.05), and deltaIC (r=0.69, p<0.05) was observed. The breathing pattern did not change significantly after bronchodilation. Stepwise multiple regression analysis of pre-BD spirometric and breathing pattern parameters selected IC and Ti/Ttot as the strongest predictors of pre-BD PEEPi dyn (p=0.039 and 0.006, respectively). CONCLUSIONS: The inhalation of 400 microg of salbutamol produced bronchodilation, reduction of air trapping, and a decrease in PEEPi dyn in spontaneously breathing patients with stable severe COPD. PMID- 16258409 TI - Apoptosis in liver diseases--detection and therapeutic applications. AB - The liver is continuously exposed to a large antigenic load that includes pathogens, toxins, tumor cells and dietary antigens. Amongst the hepatitis viruses, only hepatitis B virus (HBV) and hepatitis C virus (HCV) cause chronic hepatitis, which can progress to cirrhosis and hepatocellular carcinoma. Of the different antiviral defense systems employed by the tissue, apoptosis significantly contributes to the prevention of viral replication, dissemination, and persistence. Loss of tolerance to the liver autoantigens may result in autoimmune hepatitis (AIH). This review outlines the recent findings that highlight the role and mechanisms of apoptotic processes in the course of liver diseases. Among factors that contribute to liver pathology, we discuss the role of tumor necrosis factor (TNF)-alpha, HBx, ds-PKR, TRAIL, FasL, and IL-1alpha. Since TNF and FasL-induced hepatocyte apoptosis is implicated in a wide range of liver diseases, including viral hepatitis, alcoholic hepatitis, ischemia/reperfusion liver injury, and fulminant hepatic failure, these items will be discussed in greater detail in this review. We also highlight some recent discoveries that pave the way for the development of new therapeutic strategies by protecting hepatocytes (for example by employing Bcl-2, Bcl-XL or A1/Bfl-1, IAPs, or synthetic caspase inhibitors), or by the induction of apoptosis in stellate cells. The assessment of the severity of liver disease, as well as monitoring of patients with chronic liver disease, remains a major challenge in clinical hepatology practice. Therefore, a separate chapter is devoted to a novel cytochrome c-based method useful for the diagnosis and monitoring of fulminant hepatitis. PMID- 16258410 TI - Inherited antithrombin deficiency and end stage renal disease. AB - Antithrombin is a potent inhibitor of the coagulant effect of thrombin. In the latter half of 20th century, many families have been described in which an autosomaly dominant inherited antithrombin deficiency has caused severe venous thromboembolic disease in successive generations. The important complication is severe venoocclusive disease by deep venous thrombus. Some inherited antithrombin deficient patients developed renal failure because of fibrin deposition in the kidney glomeruli or renal vein thrombus, and therefore the need for replacement therapy for end stage renal disease (ESRD). Although an inherited antithrombin deficiency with renal failure is rare, prevention against renal failure in such patients, and their renal replacement therapy for ESRD are important. Proteinuria decreases plasma antithrombin level leading to more severe hyper-coagulation state. Therefore early in renal disease, it may be prudent for adaptation of anti coagulation therapy even if recurrent thrombosis has not occurred. All replacement therapy (hemodialysis, transplantation or peritoneal dialysis) for ESRD are available for such thrombophilic disorders. Anticoagulation agents working without aggravation of antithrombin effects (Argatroban, Nafamostat mesilate etc.) are useful for hemodialysis. The renal allograft recipients with thrombophilia seem to be at risk of developing an acute rejection or other vascular event. Peritoneal dialysis is potentially a good adaptation for such thrombophilic disorders. However which therapy has the best mortality and morbidity outcomes is not clear. Physicians and Surgeons must pay attention to the coagulation state and thrombophilia in ESRD patients, give strong consideration for adequate anti-coagulation therapy and review the best renal replacement modality for each patient. PMID- 16258411 TI - [The action of oxidative stress induced by ethanol on the central nervous system (CNS)]. AB - The brain is an organ which metabolically consumes about 20% of the total oxygen received by the organism. This causes the generation of free radicals, especially in the presence of some xenobiotics, such as ethanol. In order to prevent free radical-induced cellular damage, the organism developed a defense mechanism, the antioxidative system. The content of both exogenous and endogenous antioxidants in the central nervous system (CNS) is very small in comparison with that of other tissues, which in relation to the high level of polyunsaturated fatty acids (PUFAs) makes the CNS exceptionally susceptible to free-radical damage. The antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), and glutathione reductase (GSSG-R) are present in the CNS i.e. in the cortex, cerebellum, hypothalamus, striatum, and spinal cord, where they are responsible for the brain's basic functions, both physical and cognitive. Moreover, the highest activity of these enzymes is observed in neurons and/or glial cells. The activity of antioxidant enzymes is significantly changed in the CNS of animals chronically intoxicated with ethanol. The decrease in these activity may indicate at oxidative modification of the enzymatic proteins caused by free radicals which are generated during ethanol and acetaldehyde metabolism. It may also be caused by the decrease in the synthesis rate of these enzymes. However, the increase in the activity of antioxidant enzymes may often be explained as an adaptive reaction to an excess production of free radicals. The catalase is an exception in this respect because the decrease in its activity is related to the enhancement of protein synthesis. Ethanol intoxication also caused a decrease in GSH concentration, especially in the cerebellum, striatum, and cortex. This may be explained by the increase in the concentration of acetaldehyde, which is removed from cells with the use of this antioxidant. The antioxidative abilities of the CNS also depend on exogenous antioxidants which are provided to the organism during food intake. The most important exogenous antioxidant in the CNS is vitamin E. The content of vitamin E as well as that of vitamin C in the CNS is decreased, whereas the content of vitamin A is increased after ethanol administration. The high vitamin A level may cause damage of the central nervous system, especially in young rats exposed to ethanol in the prenatal period. PMID- 16258412 TI - [Peroxisome proliferator-activated receptors-gamma (PPAR-gamma) and their role in immunoregulation and inflammation control]. AB - Peroxisome proliferator-activated receptors-gamma (PPAR-gamma) are members of the nuclear receptor superfamily containing transcription factors regulating gene expression. PPAR-gamma have attracted attention so far as key factors in adipogenesis, lipid metabolism, insulin sensitivity, and apoptosis. Recently, growing evidence points to their implication in the regulation of the immune response, particularly in inflammation control. Not only are PPAR-gamma found in various structures of the immune system, but many inflammatory mediators, such as arachidonic acid and its metabolites, also act as potent and specific ligands of them. Inflammation is the basis of the pathogeneses of such chronic diseases as bronchial asthma, atherosclerosis, rheumatoid arthritis, and chronic inflammatory bowel diseases. The causative relationship between PPAR-gamma activity and the pathogeneses of these inflammatory disorders has been found in specific animal models. Moreover, PPAR-gamma agonists have been shown to act as potent anti inflammatory agents. Thus, PPAR-gamma can serve as potential therapeutic targets in the treatment of inflammation. The aim of this paper is to present the characteristics of PPAR-gamma regarding their gene and protein structures, ligand selectivity, mechanisms of action, and target genes. The review highlights the roles that PPAR-gamma play in inflammation and immune responses. Particular emphasis is focused on their roles in asthma, atheroclerosis, rheumatoid arthritis and chronic inflammatory bowel diseases. PMID- 16258413 TI - [Defect of glycosylation of immunoglobulin G in rheumatoid arthritis patients]. AB - Rheumatoid arthritis (RA) is a multisystem disorder in which immunological abnormalities result in symmetrical joint inflammation, articular erosion, and extra-articular involvement. The etiology of RA is still unknown, but a defect in the glycosylation of IgG may be involved in its immunopathogenesis. Several studies have shown a correlation between the amount of IgG lacking galactose and the activity of RA. IgG galactosylation has been shown to be a useful marker of early RA and an indicator of poor prognosis. Analysis of IgG galactosylation may offer an insight into disease pathogenesis and may also be useful in RA diagnosis. PMID- 16258414 TI - [The influence of metabolic changes on intrauterine fetal growth in normal gestation and gestation complicated by diabetes]. AB - Based on published clinical studies, this paper reviews the influence of metabolic changes during pregnancy on intrauterine fetal growth. Data are collected concerning the metabolic aspects of both normal pregnancies and those complicated by diabetes as well as hormonal changes having significant impact on metabolism during pregnancy. The issues of gestational insulin resistance and the metabolism of carbohydrates, lipids, proteins, and amino acids are discussed. Factors influencing birth weight and intrauterine fetal growth, including metabolic and hormonal mechanisms and the function of the placental barrier, are also considered. PMID- 16258416 TI - [Family of AMP-deaminase genes]. AB - AMP-deaminase (AMP-aminohydrolase, EC 3.5.4.6), a highly regulated oligomeric enzyme catalyzing the irreversible deamination of adenylic acid (5'-AMP), is located at a branch point of adenylate nucleotide catabolism. It plays an important role in the stabilization of adenylate energy charge (AEC) and the regulation of the purine nucleotide pool in several types of animal tissue. Tissue- and stage-specific isoforms of AMP-deaminase were described in mammals. In humans, three isozymes of AMP-deaminase, i.e. M (muscle), L (liver), and E (erythrocyte), exhibiting different physical, catalytic, and regulatory properties, were identified. AMP-deaminase activity is encoded by a multigene family in which two genes produce at least three mRNAs through alternative splicing of one of the primary transcripts. In this study we present all found and so far unpublished detailed knowledge about AMP-deaminase gene structures. We also present basic information on the effects of these gene mutations. PMID- 16258415 TI - [TRPM7: a protein responsible for magnesium homeostasis in a cell]. AB - Magnesium is an important cofactor in biological processes. For many years it has been considered that magnesium homeostasis in a cell is regulated by its eflux from a cell and not by its influx. It has also been considered that the Na+/Mg2+ antiport plays the main role. In recent years, many experiments have been carried out to understand the mechanisms of Mg2+ transport in an organism. These experiments have led to some new conclusions. It was confirmed that the level of magnesium in a cell is probably also regulated by its influx to the cell. One of the last scientific findings is the discovery of the TRPM (transient receptor potential melastatin) protein family. TRPM6 and TRPM7, bifunctional proteins with kinase and ion channel activities, are responsible for magnesium homeostasis. The discovery of these proteins led to a better understanding of magnesium homeostasis. It was confirmed that TRPM6 protein is responsible for homeostasis in the whole organism and that TRPM7 may regulate the level of magnesium in the cell. TRPM7 also has other functions. One of those newly recognized is the phosphorylation of annexin 1. However, many activities and functions of TRPM7 have not yet been described. This paper is a review of knowledge of TRPM7 transmembrane protein, which is responsible for the magnesium homeostasis in the cell. It briefly presents the main functions and structure of TRPM7. It also describes the mechanisms of its biological activity. PMID- 16258417 TI - [Chlamydia, Chlamydophila, and specific cellular immunity]. AB - In this paper studies are presented on specific cell-mediated immunity in humans and in laboratory and farm animals upon infection or immunization with bacteria of the genera Chlamydia and Chlamydophila. Such infection or immunization was demonstrated to affect the total number of T lymphocytes, their subpopulation profiles their cytotoxicity, and the activity of cytokines. The changes appeared already a few hours after infection or immunization and persisted for days or weeks. PMID- 16258418 TI - [The role of TrkC receptor and neurotrophin 3 in the development and function of neural cells]. AB - Neurotrophin 3 (NT-3) plays an important role in the development of the nervous system as well as in mediating the survival of neurons in the adult nervous system. NT-3 functions by preferential binding to the cell surface receptor TrkC as well as by binding TrkA, TrkB, and p75(NTR), with lower affinity. Various isoforms of TrkC are generated by alternative splicing. This constitutes, together with ectodomain shedding, an extensive means of regulating TrkC signaling pathways to modulate the effects of NT-3 on target cells during the differentiation of neural cells. PMID- 16258420 TI - Current evaluation of imatinib and allogeneic transplant in the treatment of chronic myeloid leukemia. PMID- 16258421 TI - New agents in colon cancer. PMID- 16258422 TI - Bone marrow transplantation in sickle cell disease: indications and successes. PMID- 16258423 TI - Transitional cell carcinoma recurrence presenting as isolated transverse sinus thrombosis. PMID- 16258425 TI - Durable local responses with subtherapeutic doses of concurrent radiation and gemcitabine in a patient with refractory Hodgkin's disease. PMID- 16258427 TI - mTOR inhibitors in hematologic malignancies. AB - Most hematologic malignancies are characterized by initial responsiveness to chemotherapeutic regimens, but many patients still die from their disease. Among the agents currently in clinical development, there is a strong rationale for evaluating rapamycin derivatives. Rapamycin was originally developed as an immunosuppressant and is approved by the US Food and Drug Administration for the treatment of kidney allograft transplant rejection. Two related compounds, RAD001 and CCI-779, are under development as cancer therapeutics. Rapamcyin binds to the immunophilin FK506-binding protein 12 and this protein/drug complex binds to and inhibits the activity of the mammalian target of rapamycin (mTOR). Among its many functions, mTOR regulates the translation of a specific subset of mRNA transcripts that encode for proteins involved in regulating the G1 to S phase transition. This review focuses on recent advances in the understanding of the mechanisms of cell growth inhibition by rapamycin, clinical trial results for the 2 agents being developed as cancer agents, and the rationale supporting the evaluation of this class of agent in hematological malignancies. PMID- 16258429 TI - A review of systemic therapy for advanced pancreatic cancer. AB - Pancreatic cancer remains an important cause of cancer mortality with few long term survivors. Improvement in the systemic therapy of pancreatic cancer is necessary to treat the frequently encountered metastatic disease. Several new chemotherapeutic agents with modest activity against pancreatic cancer have been identified over the past decade. Gemcitabine is currently the standard treatment for advanced pancreatic cancer. Combination chemotherapy trials incorporating gemcitabine, cisplatin, 5-fluorouracil, oxaliplatin, or irinotecan generally show improved outcomes in objective response rates but with little or no improvement in survival in phase III trials. Novel therapeutic strategies targeting dysregulated molecular pathways in pancreatic cancer cells are currently being explored. Future treatment regimens for pancreatic cancer will probably incorporate conventional cytotoxic drugs and novel targeted agents. PMID- 16258428 TI - Reirradiation: exploring new territory in the therapy of recurrent head and neck cancer. AB - Traditionally, systemic chemotherapy has been the main form of treatment for recurrent squamous carcinoma of the head and neck. Unfortunately, response rates are seldom higher than 20-30%, with median survival times of 6-8 months. This paper explores the emerging role of reirradiation in the setting of recurrent squamous cell carcinoma of the head and neck; it reviews the existing literature focusing on radiation alone in this setting, and more recent literature integrating systemic radiosensitizing cytotoxics with reirradiation. Data from recent series using fluorinated pyrimidines and hydroxyurea, as well as platinating agents and taxanes, suggest occasional longterm survival benefits that exceed the historic norm. These approaches, largely confined to single institutions or small consortiums to date, are being evaluated prospectively in the cooperative group setting. PMID- 16258431 TI - New drugs in acute myeloid leukemia. PMID- 16258432 TI - The promise of proteomics. PMID- 16258433 TI - Thalidomide and thrombosis. PMID- 16258434 TI - Gefitinib: current and future status in cancer therapy. AB - The epidermal growth factor receptor (EGFR) is recognized as a key modulator of tumor cell function and is considered to be a viable drug target in a range of solid malignancies. Current knowledge of its role in tumor growth and progression has led to a newly active area of anticancer research, investigating agents that target the activity of this receptor. Of these agents, gefitinib is furthest in clinical development, having received regulatory approval in Japan in 2002, and in the United States and Australia in 2003. Gefitinib is an orally active, EGFR tyrosine kinase inhibitor that blocks signal transduction pathways implicated in the proliferation and survival of cancer cells and other host-dependent processes promoting cancer cell growth. A plethora of preclinical studies have suggested promising outcomes for this agent and have led to ongoing clinical trials in a wide range of tumors, including non-small-cell lung, head and neck, colorectal, prostate, and breast, either as monotherapy or in combination with standard chemotherapy, hormonal therapy, or radiotherapy. Furthermore, as biologic agents are specifically designed to attack different pathways of tumor growth and progression, the potential for the combination of gefitinib with other agents, given either concurrently or sequentially, to prevent or delay disease recurrence is also being investigated. This article provides a detailed overview of gefitinib, the rationale for its use in a wide range of tumor types, and the current clinical development status of this novel agent. PMID- 16258435 TI - Regulation of c-Raf-1: therapeutic implications. AB - The mitogen activated protein kinase (MAPK) or Ras/Raf/MAPK kinase (MEK)/ERK signaling cascade is a ubiquitously expressed intracellular signaling pathway that transmits mitogenic stimuli to the nucleus through a series of sequential phosphorylation events and controls such cellular functions as proliferation, differentiation, and apoptosis. Components of this pathway such as ras and raf are oncogenes and as such aberrancy in their encoded proteins results in malignant transformation. The MAPK pathway is dysregulated in approximately 30% of all human tumors and therefore targeting specific components of this pathway that regulate pleiotropic cellular processes using such strategies as isoprenylation inhibitors, antisense oligodeoxyribonucleotides, and inhibitors of the kinase function represent attractive therapeutic options. raf kinase, a downstream effector of ras, has been known to be functionally aberrant in various human tumors. The 3 isoforms of raf have been studied extensively, and agents targeting c-raf are presently undergoing early-phase clinical testing. The outcomes of these trials have wide-ranging clinical implications in the management of cancers. This review addresses the rationale for targeting raf, the diverse cellular functions regulated by c-raf, and the current status of various pharmacological approaches targeting c-raf. PMID- 16258436 TI - Evolving role of irinotecan in small-cell lung cancer. AB - Irinotecan has recently been found to be one of the most active agents in the treatment of small-cell lung cancer (SCLC). Japanese investigators have led the way in the early investigation of irinotecan, and multiple studies are now ongoing in the United States. In a phase II trial conducted by the West Japan Thoracic Oncology Group, irinotecan was associated with a median survival of 13 months in patients with extensive-stage disease. Subsequently, the Japanese Clinical Oncology Group completed a phase III trial comparing irinotecan plus cisplatin to cisplatin and etoposide. In this study, median, 1-year, and 2-year survival rates were superior with irinotecan and cisplatin. Two confirmatory phase III trials are in progress in the United States. Based on these early data, it is likely that irinotecan and a platinum agent will prove to be at least as effective as any other treatment for patients with extensive-stage SCLC. Investigators have embarked on combining irinotecan with carboplatin in anticipation that this will be a preferable treatment. Phase I/II trials are complete and the doses and schedules have been recommended. As a result, we are currently exploring irinotecan and carboplatin in phase II trials in both extensive- and limited-stage settings. In addition, several of the newer biologic targeted agents are being tested in SCLC in combination with newer chemotherapy regimens. The results from these trials are eagerly awaited. PMID- 16258437 TI - Mucoepidermoid carcinoma of the lung with t(11:19)(q21:p13): a link to new biology. PMID- 16258439 TI - Cognitive functions in primary central nervous system lymphoma patients treated with chemotherapy and stem cell transplantation: preliminary findings. PMID- 16258441 TI - Therapeutic apheresis: current indication categories. PMID- 16258442 TI - Finding the right dose. PMID- 16258443 TI - Profiling in acute leukemia. PMID- 16258444 TI - Sentinel lymph node biopsy. PMID- 16258445 TI - Adjuvant chemotherapy for non-small-cell lung cancer. AB - Survival rates following complete resection for patients with non-small-cell lung cancer are disappointing. Only 60-70% of patients with stage I disease (no lymph node involvement) are expected to survive 5 years. Attempts to improve survival have included the use of chemotherapy, radiation, or both before or after surgery. The majority of randomized trials examining the use of postoperative therapies have not found a survival benefit. Many of these trials have enrolled small numbers of patients and have been underpowered to detect small, but significant, survival differences. Recent data from large, randomized international trials have yielded conflicting results. The use of postoperative therapy should continue to be studied in clinical trials and a new meta-analysis incorporating results from recently completed randomized trials should be conducted. PMID- 16258446 TI - Strategies for the treatment of imatinib-resistant chronic myeloid leukemia. AB - Treatment of chronic myeloid leukemia (CML) patients using imatinib alone is unlikely to be curative. The challenges in the era of imatinib are to prevent the emergence of imatinib resistance and to identify the most effective alternative approaches for patients who lose imatinib responsiveness. This review describes the possible strategies to overcome imatinib resistance in CML patients based on the current understanding of the action of imatinib and mechanisms of its resistance. The useful laboratory tests to study imatinib resistance and a current Australian CML study employing imatinib dose intensification and sequential combination therapy for newly diagnosed patients are also outlined. PMID- 16258447 TI - Low-grade glioma: current management and controversies. AB - The question of optimal management of patients with low-grade gliomas continues to generate considerable controversy. Retrospective studies have helped delineate prognostic factors for patients with these tumors as well as the relative efficacy of surgery, radiation and chemotherapy. Results of recently completed and ongoing prospective, multi-institutional trials will help guide the treatment of these patients in the future. Other advances include the molecular characterization of these tumors that should assist in selection of existing and development of future therapies. PMID- 16258448 TI - Non-Hodgkin's lymphoma occurring as pure central nervous system pathology. PMID- 16258451 TI - New concepts in tissue-specific metastases. PMID- 16258452 TI - Are there any better camptothecins than the ones we have? PMID- 16258453 TI - Current management and new approaches in the treatment of APL. PMID- 16258454 TI - Aging and coagulation. PMID- 16258455 TI - Current treatment recommendations for stage III non-small-cell lung carcinoma. AB - Nearly one third of patients with non-small-cell lung cancer (NSCLC) present with stage III disease. In general, these patients have a dismal prognosis. However, several studies discussed in this review suggest that aggressive therapy in appropriately selected patients can improve outcomes. These studies can be difficult to interpret, not only because of the diversity of patient populations studied, but also because of the variability in eligibility criteria, pretreatment staging techniques, and induction regimens used. Nonetheless, sound treatment recommendations can be made based on the available literature. Ongoing research is likely to clarify further the best combination of surgery, chemotherapy, radiation, and novel biologic agents to optimize treatment for this heterogeneous population. This review will focus primarily on the influential prospective randomized trials, which have shaped current treatment practices for stage III NSCLC. PMID- 16258456 TI - Bortezomib: a novel therapy approved for multiple myeloma. AB - Cellular homeostasis requires routine degradation of key regulatory proteins, including tumor suppressor gene products, transcription factors, cell-cycle proteins and their inhibitors, as well as damaged and misfolded proteins. A critical part of this process is mediated by the 26S proteasome, a multi-subunit enzyme found in the nucleus and cytoplasm of all eukaryotic cells. Because of its essential role in many cellular processes controlling growth and survival, the proteasome has been identified as a potential target for cancer therapy. Drugs known to inhibit proteasome activity have been shown to induce cell-cycle arrest and programmed cell death (apoptosis). The impact of this finding is heightened by research showing that cancer cells are more sensitive to the proapoptotic effects of proteasome inhibition than normal cells. Preclinical evidence using bortezomib, the only proteasome inhibitor to enter clinical trials, suggests that proteasome inhibition may be effective in the treatment of hematologic and solid malignancies by promoting apoptosis, retarding angiogenesis, and inhibiting tumor cell adhesion and production of growth factors by acting on molecules such as nuclear factor-kappaB. Further preclinical evidence suggests that the antitumor effects of cytotoxic chemotherapy or radiotherapy may be enhanced by the addition of a proteasome inhibitor. Bortezomib was recently approved for the treatment of multiple myeloma. It is currently being investigated, both as a single agent and in combination, in phase I and II trials in a variety of tumor types. PMID- 16258457 TI - Participation in cancer trials: recruitment of underserved populations. AB - One approach to address cancer health disparities is to focus on the under representation by minority populations in cancer trials. Recruitment strategies include: 1) characterizing the target populations, 2) involve members of the population in planning, 3) take the message to the population, 4) give something back to the community, 5) enhance credibility with a community spokesperson, 6) identify and remove barriers, 7) improve staff sensitivity, and 8) educate the population about the trial. To recruit minorities to clinical trials, we have developed the Accrual to Clinical Trials (ACT) framework for understanding and enhancing the recruitment of participants to cancer trials. PMID- 16258458 TI - Extranodal rectal Hodgkin's lymphoma in an HIV-positive individual. PMID- 16258459 TI - Hodgkin's lymphoma in HIV: unusual presentations and improving outcomes. PMID- 16258460 TI - Genetics in the management of adult de novo AML. PMID- 16258461 TI - Developing targeted therapies for lung cancer. PMID- 16258462 TI - Developing drugs that do not cause tumor regression. PMID- 16258463 TI - Warfarin sensitivity and genetic polymorphisms: should pharmacokinetic screening be part of anticoagulant dosing decisions? PMID- 16258466 TI - Polycythemia vera with uncommon presentations. PMID- 16258464 TI - Lymphomatoid granulomatosis treated with rituximab and chemotherapy. PMID- 16258468 TI - Cancer and thrombosis. AB - Cancer is frequently complicated by thrombosis, which may even be the first sign of an unrecognized cancer. There are several risk factors for cancer-associated thrombosis, such as type of malignancy, the presence of metastatic disease, and the use of chemotherapy. In most patients with cancer, a procoagulant state can be identified, ranging from subclinical laboratory abnormalities to full-blown disseminated intravascular coagulation. A specific entity is the occurrence of thrombotic microangiopathy that is specifically related to (high-dose) chemotherapy and radiotherapy. The pathogenetic pathways that play a role in the cancer-associated coagulopathy have been identified in recent years. PMID- 16258469 TI - Clotting mechanisms and cancer: implications in thrombus formation and tumor progression. AB - Development of cancer is associated with activation of blood coagulation. The results of laboratory tests clearly demonstrate that fibrin formation and dissolution is continuously ongoing at different rates in these patients, who are at increased risk of secondary thrombosis. Notably, fibrin formation is also involved in the process of tumor spread and metastasis. The pathogenesis of the hemostatic disorders in cancer is complex and reflects the interaction of different mechanisms involving the activation of various hemostatic components, such as the coagulation and fibrinolytic systems, the vascular endothelium, leukocytes, and platelets. Tumor cells possess the capacity to interact with all of these components. Indeed they directly activate the coagulation cascade by producing their own procoagulant factors, or they can stimulate the prothrombotic properties of other blood cell components. Additional mechanisms of blood clotting activation are started by the initiation of antitumor therapies. In the last 10 years research studies have greatly improved our knowledge of tumor promoted prothrombotic functions. Understanding the molecular basis of the underlying mechanisms may help to identify better-targeted strategies to prevent thromboembolism in cancer patients. Further, pharmacological modulation of malignant cell hemostatic properties may not only affect the tumor-associated thrombotic risk but may also leave open the possibility to interfere with the progression of the disease. PMID- 16258470 TI - Optimizing the management of chemotherapy-induced neutropenia. AB - Chemotherapy-induced neutropenia (CIN), the most common dose-limiting toxicity of cancer chemotherapy, is associated with numerous clinical, personal, and economic consequences. The principal strategies for managing CIN are reducing the dose intensity of the chemotherapy, using antibiotics, and using colony-stimulating factors (CSFs). Reducing or delaying the chemotherapy dose is effective, but this can compromise treatment outcomes. Antibiotics can be lifesaving, but they are associated with numerous adverse effects and the emergence of resistant pathogens. The granulocyte CSF (G-CSF) filgrastim can reduce the incidence, duration, and severity of CIN, as well as the risk of infection, in patients treated with myelosuppressive chemotherapy. The availability of pegfilgrastim, a sustained-duration G-CSF that has benefits comparable to those of filgrastim with a single injection per chemotherapy cycle, has simplified CSF therapy. PMID- 16258472 TI - New therapies for mantle cell lyphoma. PMID- 16258474 TI - Complications of chronic venous access devices in cancer patients. PMID- 16258473 TI - Current developments in treating chemotherapy-related oral mucositis. PMID- 16258475 TI - Are targeted therapies really targeted? PMID- 16258476 TI - Antiphospholipid antibodies. AB - Antiphospholipid antibodies (APAs) have been associated with thromboembolic events. Since they were fi rst described in 1906, APAs have been the subject of multidisciplinary studies seeking to link them to potential pathophysiologic mechanisms. This review summarizes the different types of APAs, antigenic targets, clinical complications associated with APAs, and APA syndrome. In addition, the currently available methods for laboratory identification of lupus anticoagulants are discussed, as is the laboratory diagnosis of anticardiolipin antibodies. PMID- 16258477 TI - The role of prophylactic cranial radiation in the treatment of non-small-cell lung cancer. AB - Patients with lung cancer face a substantial risk of developing brain metastases. Prophylactic cranial irradiation therapy has been demonstrated to significantly decrease the incidence of central nervous system metastasis and improve overall survival in patients with small-cell lung cancer. In non-small-cell lung cancer, the central nervous system is a common site of systemic failure and frequently represents the only focus of extra-thoracic disease. However, the role of prophylactic cranial irradiation in patients with non-small-cell lung cancer is less well defined. As treatment modalities improve and patients live longer their risk of developing central nervous system recurrence increases, and methods to prevent this demand exploration. PMID- 16258478 TI - Sphincter preservation therapy for rectal cancer. AB - Rectal cancer presents a unique challenge to oncologists and patients due to the location and anatomy of the rectum and the difficulties inherent in pre-operative staging. These issues are especially important with distal rectal tumors when patients may face the decision of tumor control without sphincter preservation or more limited surgical procedures that may potentially compromise tumor control and thus survival. Current options for sphincter preservation for low-lying rectal tumors are preoperative radiotherapy with or without chemotherapy for tumor downstaging, local excision with or without adjuvant chemoradiation and low anterior resection with coloanal anastomosis. Pretreatment evaluation, by radiologic studies and pathologic predictors of lymph node involvement, is an integral part of determining which patients are suitable candidates for treatment with local excision. Preoperative chemoradiotherapy is a treatment option for some patients who are not initially considered to be candidates for sphincter preservation. Many investigators have suggested that the rate of sphincter preservation in patients with rectal cancer may be improved following preoperative chemotherapy and radiation. For properly selected patients, local excision holds promise as a means of achieving sphincter preservation. PMID- 16258482 TI - New targets in oncology drug development. PMID- 16258479 TI - Neoadjuvant chemoradiation to convert locally advanced pancreatic body adenocarcinoma to resectable disease. PMID- 16258483 TI - Update on the use of imatinib mesylate. PMID- 16258484 TI - Chemoprevention of head and neck cancer. PMID- 16258485 TI - Coagulation and thrombotic complications associated with ventricular assist devices. PMID- 16258486 TI - Current management of oral mucositis. PMID- 16258487 TI - Treatment of zygomycosis with posaconazole in a patient with acute myeloid leukemia. PMID- 16258488 TI - Review of treatment of zygomycosis with posaconazole in a patient with acute myeloid leukemia. PMID- 16258489 TI - Update on new diagnostic and therapeutic approaches for sarcomas. AB - Sarcomas represent a family of uncommon malignancies related by histopathology and developmental biology. Sarcomas arise from any of the mesodermal tissues anywhere in the body (muscles, tendons, adipose tissue, blood vessels, and joints) from cells of mesenchymal origin or in the peripheral nervous system, derived from the ectoderm. Mainly diagnosed in the extremities, sarcomas can be present also in soft tissues of the trunk, abdomen, and retroperitoneum, as well as the head and neck. During the last decade, improvements in diagnostic techniques have made it possible to identify and characterize a subset of sarcomas arising within the gastrointestinal tract known as gastrointestinal stromal tumors, which were previously commonly misclassified as leiomyosarcomas. Sarcomas are extremely heterogeneous, so expert histopathologic characterization is essential to choose the appropriate path of multidisciplinary treatment and predict possible clinical outcomes. The aim of this review is to provide a brief overview of the epidemiology, pathology, clinical presentation, and new approaches to the treatment of adult soft-tissue and bone sarcomas. PMID- 16258490 TI - Current roles for allogeneic hematopoietic cell transplantation following nonmyeloablative or reduced-intensity conditioning. AB - Nonmyeloablative and reduced-intensity conditioning regimens followed by allogeneic hematopoietic cell transplantation (HCT) have been evaluated in patients with hematologic malignancies who were not considered candidates for conventional HCT because of age or medical comorbidities and in selected patients with metastatic renal cell carcinoma. The regimens have relied more on graft versus-tumor effects than on chemoradiation therapy to facilitate engraftment and eradicate malignant cells. While nonmyeloablative HCT has been associated with reduced regimen-related toxicities and has been curative for a number of patients with hematologic malignancies, challenges have remained in regard to graft-versus host disease, infections, and disease progression. In this article, we review data from a number of published phase I and II studies that describe the results of allogeneic HCT after nonmyeloablative conditioning. PMID- 16258492 TI - Chemoprevention of melanoma: theoretical and practical considerations. AB - BACKGROUND: Chemoprevention refers to the use of agents to reverse, suppress, or prevent carcinogenic progression of cancer. The use of chemoprevention is an unexplored strategy in melanoma prevention. METHODS: A retrospective review of the literature was undertaken regarding the important elements in evaluating chemoprevention as a strategy in melanoma. RESULTS: Several considerations need to be addressed before a chemoprevention agent can be moved to a large randomized trial. Statins have both experimental and epidemiologic evidence to support their further development as candidate chemopreventive agents, but the evidence is insufficient to justify large-scale phase III studies. A strong scientific rationale, a systematic approach to chemoprevention agent development with rigorous chemoprevention designs, and careful selection of surrogate endpoint biomarkers are critical issues in developing a chemoprevention strategy. CONCLUSIONS: Addressing these relevant considerations will allow for the development of chemoprevention in melanoma. PMID- 16258493 TI - Cutaneous melanoma: prognostic factors. AB - BACKGROUND: Recent data have changed our views of prognostic factors in cutaneous melanoma. While some newer methods have yielded better prognostic information, some insights have evolved as a result of large-scale population-based analyses. METHODS: We review current data on several different prognostic factors and divide these factors according to their application in localized primary melanoma or metastatic melanoma. For each prognostic factor, the level of evidence supporting its use and its applicability to clinical practice are considered. RESULTS: For localized primary melanoma, the dominant predictors of survival include lesion thickness, ulceration, and lymph node involvement. Factors such as age, sex, anatomic location, and satellite/in-transit lesions are important in localized melanoma. Factors currently being investigated are tumor vascularity, vascular invasion, mitotic rate, tumor regression, and tumor-infiltrating lymphocytes. For metastatic melanoma, the most important prognostic factors are site of metastases and the presence of elevated serum lactic dehydrogenase. The value of these prognostic factors to clinicians caring for melanoma patients is discussed. CONCLUSIONS: A better understanding of prognostic factors in cutaneous melanoma has evolved over the last decade, allowing oncologists to provide appropriate treatment for their patients. Many of the prognostic factors are interrelated. In the near future, it is expected that several molecular genetic factors will provide more insight into the prognosis of patients with melanoma. PMID- 16258494 TI - Sentinel node biopsy for thin melanomas: which patients should be considered? AB - BACKGROUND: As the incidence of melanoma increases, thin melanomas are being diagnosed at an increasingly frequent rate. Currently available prognostic factors are limited in their ability to reliably discriminate which patients will manifest regional nodal metastasis and would be identified early through sentinel node biopsy. METHODS: We summarized our experience with sentinel node biopsy for patients with cutaneous melanomas less than 1.00 mm in Breslow thickness, with evaluation of Clark level as a predictor of positive sentinel node metastasis. RESULTS: Among the 409 patients identified, micrometastases were found in the sentinel node in 20 patients, for an overall incidence of nodal progression of 4.9%. A total of 252 (62%) were Clark level II or III (11 of whom had a positive sentinel node) and 157 (38%) were Clark level IV (9 of whom had a positive sentinel node). We reviewed the literature to identify reliable indicators that might be helpful in determining which patients with "thin melanomas" would be likely to manifest regional progression to warrant routinely undergoing a preoperative lymphoscintigraphy followed by a sentinel node biopsy. CONCLUSIONS: Based on available data, patients with melanomas between 0.75 and 1.00 mm are appropriate candidates to be considered for sentinel node biopsy after discussing the likelihood of finding evidence of nodal progression, the risks of sentinel node biopsy (including the risk of a false-negative result), and the lack of proven survival benefit from any form of surgical nodal staging. PMID- 16258496 TI - Head and neck melanoma. AB - BACKGROUND: Melanoma of the head and neck and its treatment are complex issues. The behavior of head and neck melanoma is aggressive, and it has an overall poorer prognosis than that of other skin sites. METHODS: The authors review current data on the treatment of head and neck melanoma, including both cutaneous and mucosal melanoma. RESULTS: Current understanding of the behavior of head and neck melanoma is reviewed and treatment stratagems are presented. Controversies in treatment include lymphoscintigraphy with sentinel node biopsy, nodal dissection, margin size, role of radiation therapy, and reconstruction. The management goal is to treat melanoma aggressively while minimizing the effects of treatment on patient quality of life. CONCLUSIONS: Due to its aggressiveness, head and neck melanoma should be treated aggressively when morbidity is not significantly increased. Patient specific treatment is imperative. PMID- 16258495 TI - Choices in adjuvant therapy of melanoma. AB - BACKGROUND: High-dose interferon (IFN) is the approved agent for adjuvant treatment of melanoma in the United States. This approval is for high-risk, predominantly stage III patients with cutaneous primaries. There are still decisions to be made in the care of these patients. Also, there are questions about whether the IFN data can be extrapolated to patients with other stages of melanoma and whether adjuvant treatment should be offered to these individuals. Clearly there is room for improvement in this area. METHODS: The literature on this topic and ongoing national trials in the United States were reviewed. RESULTS: The data are insufficient to recommend other agents in the adjuvant treatment of melanoma outside a clinical trial. Extrapolation of the IFN data to patient populations other than those studied is problematic at best. National trials are available for most patient populations. CONCLUSIONS: The adjuvant treatment of choice for melanoma patients is participation in a clinical trial. PMID- 16258497 TI - Enhanced detection of hepatocellular carcinoma. AB - BACKGROUND: Tumor markers in the early detection of tumors are promising tools that could improve the control and treatment of tumors. While alpha-fetoprotein (AFP) is a commonly used tumor marker in the detection of hepatocellular carcinoma (HCC), its sensitivity and specificity are insufficient to detect HCC in all patient samples. METHODS: We compared AFP with serum levels of vascular endothelial growth factors (VEGF and VEGF-A), insulin-like growth factor-2 (IGF II), and the activity of the lysosomal enzyme alpha-L-fucosidase (AFU) in the sensitivity of detection of HCC and cirrhosis in Egyptian patients. RESULTS: The sensitivity of tumor detection using AFP was 68.2%. This level of detection was increased to 88.6% when AFP was evaluated in conjunction with AFU. The combined use of AFP and VEGF increased the sensitivity of detection to 95.5% in patients with HCC. The combination of the three markers yielded 100% detection sensitivity. VEGF-A showed a low specificity (20%), and IGF-II showed extremely low sensitivity (4.5%). CONCLUSIONS: We suggest that AFU or VEGF or both be measured with AFP to improve the detection sensitivity of HCC. PMID- 16258498 TI - Review of fluorine-18-2-fluoro-2-deoxy-D-glucose positron emission tomography (FDG-PET) in the follow-up of medullary and anaplastic thyroid carcinomas. AB - BACKGROUND: The goal of posttreatment follow-up for medullary and anaplastic thyroid cancer (MTC and ATC) is the early diagnosis of recurrence or metastases. However, routine follow-up protocols, including physical examination and clinically oriented investigations, are not standardized, and their sensitivity in accurately detecting recurrent or metastatic disease is often suboptimal. A valuable addition to posttreatment follow-up of oncology patients is positron emission tomography using fluorine-18-2-fluoro-2-deoxy-D-glucose (FDG-PET). METHODS: We review the role of FDG-PET imaging in the follow-up of patients previously treated for MTC and ATC. RESULTS: Based on the encouraging literature data, FDG-PET appears to be useful in detecting recurrent or metastatic disease in patients with MTC and ATC, providing a higher sensitivity (66% to 100%) and specificity (79% to 90%) than conventional imaging methods. However, the PET technique is limited by less accurate spatial assignment of small lesions, especially in the lung and liver. CONCLUSIONS: Supporting evidence indicates that FDG-PET has a significant role in the follow-up of patients with MTC and ATC. PMID- 16258499 TI - Histone H3.3 deposition at E2F-regulated genes is linked to transcription. AB - The histone variant H3.3 can be incorporated in chromatin independently of DNA synthesis. By imaging using green fluorescent protein-tagged histones, H3.3 deposition has been found to be linked with transcriptional activation. Here, we investigated H3.3 incorporation during G1 progression on cell-cycle-regulated E2F dependent genes and on some control loci. We transiently transfected resting cells with an expression vector for tagged H3.3 and we analysed its presence by chromatin immunoprecipitation. We found that replication-independent H3.3 deposition occurred on actively transcribed genes, but not on silent loci, thereby confirming its link with transcription. Interestingly, we observed similar levels of H3.3 occupancy on promoters and on the coding regions of the corresponding genes, indicating that H3.3 deposition is not restricted to promoters. Finally, H3.3 occupancy correlated with the presence of transcription competent RNA polymerase II. Taken together, our results support the hypothesis that H3.3 is incorporated after disruption of nucleosomes mediated by transcription elongation. PMID- 16258500 TI - Dual-color, break-apart fluorescence in situ hybridization for EWS gene rearrangement distinguishes clear cell sarcoma of soft tissue from malignant melanoma. AB - Clear cell sarcoma of soft tissue (malignant melanoma of soft parts) is a soft tissue sarcoma with melanocytic differentiation that typically occurs in the tendons and aponeuroses of young adults. As demonstrated by cytogenetics and reverse-transcriptase polymerase chain reaction, between 70% and over 90% of clear cell sarcomas have a t(12;22) translocation, fusing the EWS and ATF1 genes on chromosomes 22q12 and 12q13, respectively. Identification of this translocation distinguishes clear cell sarcoma from histologic mimics, most importantly conventional malignant melanoma. We report our experience with a commercially available, dual-color, break-apart fluorescence in situ hybridization (FISH) probe, which allows detection of EWS (22q12) gene rearrangement in formalin-fixed, paraffin-embedded tissues. Histologically and immunophenotypically well-characterized cases of clear cell sarcoma (n = 10) and malignant melanoma (n = 32) were evaluated with a 22q12 dual-color, break-apart probe (Vysis, Downer's Grove, IL, USA), which spans the known common breakpoints in the EWS gene on chromosome 22 (introns 7-10). Signals from tumor cell nuclei were counted under a fluorescence microscope and the presence of red-green break apart signals was recorded. Of the clear cell sarcoma cases, seven of 10 showed evidence of an EWS gene rearrangement with a mean of 81.6% positive cells per sample (range: 60-95%). All cases of malignant melanoma (n = 32) showed virtually absent break-apart signals in the EWS gene (less than 4% cells per case). FISH detects EWS gene rearrangement in a substantial proportion of clear cell sarcomas, with excellent specificity. Importantly, EWS FISH is negative in malignant melanoma, a clinically dissimilar tumor, which may closely mimic clear cell sarcoma histologically and immunohistochemically. As the studied probe can be utilized in routinely processed tissue, FISH provides an excellent alternative to reverse-transcriptase polymerase chain reaction in cases where fresh tissue is unavailable. PMID- 16258501 TI - DNA hypermethylation status of multiple genes in soft tissue sarcomas. AB - The aberrant methylation of promoter CpG islands is known to be a major inactivation mechanism of tumor-related genes. To determine the clinicopathological significance of gene promoter methylation in soft tissue sarcomas, we examined the promoter methylation status of 10 tumor-related genes in 65 soft tissue sarcomas and 19 adjacent non-neoplastic tissues by methylation specific PCR. The methylation frequencies of tumor-related genes tested in soft tissue sarcomas were 17 (26%) for RASSF1A, 11 (17%) for DAP kinase, 10 (15%) for MGMT, nine (14%) for GSTP1, eight (12%) for PTEN, six (9%) for p16 and hMLH1, five (8%) for hMSH2, two (3%) for p14, and one (2%) for RB. Promoter methylation of these genes was not recognized in non-neoplastic tissues. All those cases of soft tissue sarcoma that had MGMT methylation, with the exception of one case of malignant peripheral nerve sheath tumor, showed large tumor size (> or = 10 cm) or recurrence. Moreover, eight of 10 cases with MGMT methylation revealed high American Joint Committee on Cancer stage. Seven of 10 cases (70%) with MGMT methylation showed a loss of MGMT expression by immunohistochemistry. In addition, MGMT methylation status had a statistically significant correlation with a loss of MGMT expression (P=0.014). In conclusion, although methylation of tumor-related genes was a relatively rare event in soft tissue sarcomas, methylation was tumor-specific. Of 10 tumor-related genes, cases with MGMT methylation had a tendency to be aggressive behavior. Moreover, MGMT methylation was closely associated with a loss of MGMT expression. Although our findings need to be extending to a large series, promoter methylation of tumor-related genes is likely to have an association with the pathogenesis of soft tissue sarcomas. Furthermore, MGMT methylation may be associated with tumor aggressiveness and the inactivation of MGMT gene. PMID- 16258502 TI - Hodgkin's lymphoma of the thyroid: a clinicopathologic study of five cases and review of the literature. AB - Hodgkin's lymphoma rarely involves the thyroid gland. We report the clinical and pathologic features of five cases of Hodgkin's lymphoma that presented as thyroid lesions. All five patients were females, aged 29-59 years. Three patients had a history of chronic thyroiditis and hypothyroidism and two were euthyroid. One patient had a remote history of Hodgkin's lymphoma. Imaging studies showed a 'cold' nodule (three cases) or a diffusely enlarged thyroid gland, resembling goiter or fibrosclerosing thyroiditis (two cases). Thyroid fine-needle aspiration was performed before thyroidectomy in all cases; three of these cases contained some atypical cells, raising the possibility of Hodgkin's lymphoma. Histologically, all cases were classical Hodgkin's lymphoma, nodular sclerosis subtype. The four patients with primary thyroid lymphoma had Stage IIE disease. All patients were treated with surgical excision and chemotherapy, with or without radiation therapy, and were alive after 2 months to 7 years of follow-up. A review of the English literature between 1962 and 2005 revealed 16 cases of thyroid Hodgkin's lymphoma, with a female preponderance and generally favorable outcome similar to the cases in our series. Hodgkin's lymphoma of the thyroid is rare and can mimic a primary thyroid epithelial tumor or thyroiditis clinically. Histologic diagnosis may be difficult due to marked fibrosis. Hodgkin's lymphoma should be considered in the differential diagnosis of thyroid neoplasms. PMID- 16258503 TI - Lymphoid neoplasms associated with concurrent t(14;18) and 8q24/c-MYC translocation generally have a poor prognosis. AB - We identified 14 B-cell neoplasms with concurrent t(14;18) and chromosome 8q24 or c-MYC translocations shown by conventional cytogenetics or fluorescence in situ hybridization analysis. All cases assessed by conventional cytogenetics had a complex karyotype. There were 10 men and four women, with a median age of 55 years (range, 29-72). None of these patients had a history of follicular lymphoma. The biopsy specimens were obtained from bone marrow, lymph node, and extranodal sites. Morphologically, nine neoplasms had features of Burkitt or atypical Burkitt lymphoma/leukemia and three were diffuse large B-cell lymphoma with high-grade cytologic features. The remaining two cases were plasmablastic myeloma and low-grade B-cell lymphoma, respectively. All cases expressed BCL-2. The proliferation index assessed by using Ki-67 (MIB1) was 5% in the low-grade B cell lymphoma, 80% in the plasmablastic myeloma, 90-95% in three cases of diffuse large B-cell lymphoma, and ranged from 90 to >99% in most Burkitt and atypical Burkitt neoplasms. The patient with low-grade B-cell lymphoma was treated with rituximab. All other patients received intensive combination chemotherapy. Two of these patients underwent bone marrow transplantation, and one patient received radiation therapy in addition to transplantation. The median follow-up period was 9 months (range, 3-81). In all, 10 patients died with a median survival of 9 months (range, 3-81). We conclude that most B-cell lymphomas with concurrent t(14;18) and 8q24/c-MYC translocations fall within the morphologic spectrum of diffuse large B-cell and Burkitt lymphoma. These neoplasms are high-grade and are associated with a poor prognosis. However, this combination of molecular abnormalities can also rarely occur in other neoplasms, such as the cases of low grade B-cell lymphoma and plasmablastic myeloma in this study. PMID- 16258504 TI - Genomic aberrations in mucinous tubular and spindle cell renal cell carcinomas. AB - Mucinous tubular and spindle cell carcinoma of the kidney is a new diagnostic entity. We present the pathologic and genomic characteristics of three such low malignant tumors. Two of the tumors were found in women aged 19 and 52 years, the third tumor was found in an 80-year-old man, and the tumor stages were pT2N0MX, pT2NXMX, and pT1NXMX, respectively. Findings by immunohistochemistry were similar but not identical for the three cases; markers for both proximal and distal parts of the nephron were expressed in each tumor, a finding that is in agreement with data from previous studies. The Ki-67-labeling index was below 5 in all three cases. Two of the tumors were predominantly hypodiploid (DNA-indexes 0.77 and 0.80), whereas the third tumor was hypertriploid (1.57) as measured by DNA-image cytometry. From the latter tumor live cells were available making it possible to establish its karyotype: 62-70,XXX,+del(X)(q11),-1,+2,+4,-5,-6,+7,-8,-9,-10, 11,+12,-13,-14,-15,+16,+17,+18,-19,+20,+21,-22[cp15]. Interphase fluorescence in situ hybridization analyses with centromere-specific probes for chromosomes 1, 3, 4, 6, 7, 9, 10, 17, 18, 20, and X showed that the two hypodiploid tumors had disomic and monosomic chromosome populations, whereas the karyotyped, near triploid tumor was dominated by trisomic chromosome populations. Comparative genomic hybridization analysis was normal for the karyotyped tumor but abnormal for the two others. We conclude that multiple numerical chromosome aberrations may be a feature of mucinous tubular and spindle cell carcinomas of the kidney, but beyond that no clear-cut karyotypic aberration pattern is so far discernible. PMID- 16258505 TI - Eosinophilic esophagitis in adults: distinguishing features from gastroesophageal reflux disease: a study of 41 patients. AB - Eosinophilic esophagitis in adults is a recently described entity occurring in young males with dysphagia, in whom esophageal biopsies show eosinophilic infiltration. This study defines the clinical and histological features of patients with eosinophilic esophagitis, distinguishing it from gastroesophageal reflux disease. Esophageal biopsies from patients with dysphagia or esophagitis were reviewed blindly, and assessed for: epithelial eosinophil counts, presence of eosinophilic microabscesses, edema, basal zone hyperplasia, lamina propria papillae elongation, eosinophils and fibrosis. Clinical and endoscopic findings were obtained. Eosinophilic esophagitis was diagnosed with epithelial eosinophils > or = 15 in > or = 2 high-power fields (hpfs) or > or = 25 in any hpf. Analysis was performed with Mann-Whitney, chi2 and ANOVA tests. Of 157 cases, 41 had eosinophilic esophagitis. Male gender (81%) and age < or = 45 (54%) were commoner in patients with eosinophilic esophagitis (P = 0.001, 0.010, respectively). Dysphagia was more common in eosinophilic esophagitis patients (63%, P < 0.001); heartburn was more common in noneosinophilic esophagitis patients (53%, P < 0.001). Endoscopic rings were more common in eosinophilic esophagitis patients (27%, P = 0.023); hiatus hernia was more common in noneosinophilic esophagitis patients (11%, P = 0.022). Eosinophils were more numerous in eosinophilic esophagitis biopsies (mean 39/hpf, P < or = 0.001). Only eosinophilic esophagitis biopsies had eosinophilic microabscesses (42%, P < or = 0.001). Edema, basal zone hyperplasia, lamina propria papillae elongation and lamina propria eosinophils were commoner in eosinophilic esophagitis (P < or = 0.001-0.002), while lamina propria fibrosis was specific for eosinophilic esophagitis (39%, P < 0.001). Eosinophilic esophagitis is a disease with a predilection for young males with dysphagia and rings on endoscopy. Biopsies in eosinophilic esophagitis have high epithelial eosinophil counts, averaging nearly 40/hpf. Increased awareness of eosinophilic esophagitis is necessary, since treatment with allergen elimination or anti-inflammatory therapy may be more effective than acid suppression. PMID- 16258506 TI - Loss of expression and nuclear/cytoplasmic localization of the FOXP1 forkhead transcription factor are common events in early endometrial cancer: relationship with estrogen receptors and HIF-1alpha expression. AB - The FOXP1 gene has been identified as a new member of the winged helix family of transcription factors that have important roles in cellular transformation, differentiation and proliferation. In this study, we examined the expression of FOXP1 in the normal and malignant endometrium (stage I endometrioid adenocarcinoma cases), showing a frequent deregulation of its expression in cancer. Proliferative endometrium showed predominantly nuclear localization of FOXP1, while exclusively weak cytoplasmic staining was present in the secretory phase. Loss of nuclear expression was the most striking event in endometrial adenocarcinoma. Nuclear expression ranged from 0 to 20% (median 0%). Cytoplasmic expression was noted more frequently, ranging from 0 to 90% of cancer cells (median 30%). Overall, 24/82 cases (29.3%) were observed to lack both nuclear and cytoplasmic FOXP1 expression. Tumors with exclusively cytoplasmic expression of FOXP1 were linked with deep myometrial invasion and hypoxia-inducible factors 1alpha (HIF-1alpha) expression. On the other hand, the presence of nuclear FOXP1 expression was significantly linked with ER-alpha reactivity. Survival analysis did not reveal significant differences among patients grouped by FOXP1 expression, presumably due to the high curability of stage I disease. This study provides evidence on pathways to be investigated to elucidate the interplay between FOXP1, ER-alpha and HIF-1alpha in hormone dependent cancers. PMID- 16258507 TI - Expression of hepatocyte nuclear factor-1beta (HNF-1beta) in clear cell tumors and endometriosis of the ovary. AB - Clear cell tumors of the ovary are frequently associated with ovarian endometriosis. Clinicopathologically, it has been suggested that clear cell tumors develop from endometriosis, but there has been little molecular evidence supporting this speculation. Microarray analysis revealed recently that hepatocyte nuclear factor-1beta (HNF-1beta) was significantly upregulated in clear cell carcinoma of the ovary. In the present study, we examined 30 clear cell tumors (26 malignant, three borderline, and one benign) and 40 endometriotic cysts to clarify if differentiation into the clear cell lineage already begins in ovarian endometriosis. All of the 30 clear cell tumors, including borderline and benign ones, showed immunohistochemical expression of HNF-1beta in the nucleus, while other types of ovarian epithelial tumors (endometrioid, serous, mucinous, and Brenner tumors) rarely expressed it. Among 30 clear cell tumors, 17 (56%) cases were associated with endometriosis, and endometriotic epithelium was identified in 12 cases. In nine of the 12 cases, distinct nuclear immunostaining for HNF-1beta was detected in the endometriotic epithelium, as well as in the clear cell tumor. HNF-1beta expression was observed either in atypical endometriosis (four cases), or in endometriosis of a reactive nature (five cases). Furthermore, 16 of 40 (40%) endometriotic cysts without a neoplasm also expressed HNF-1beta, and the expression was almost exclusively observed in the epithelium showing inflammatory atypia. Our results indicate that HNF-1beta is an excellent molecular marker for ovarian clear cell tumors, including benign, borderline and malignant lesions. Early differentiation into the clear cell lineage takes place in ovarian endometriosis, not only in atypical endometriosis, but also in endometriosis with degenerative and regenerative changes, and this is probably responsible for the frequent occurrence of clear cell carcinoma in ovarian endometriosis. PMID- 16258508 TI - TMA-Combiner, a simple software tool to permit analysis of replicate cores on tissue microarrays. AB - We have previously published a suite of software tools that facilitates the reformulation of tissue microarray (TMA) data so that it may be analyzed using techniques originally devised for analysis of cDNA microarray data. However, current microarray data often feature multiple scores for a given tissue sample and antibody combination. Furthermore, an efficient and systematic method for combining scores that takes into account the differing staining properties of tissue epitopes has not been described. We thus present the TMA-Combiner, a new Microsoft Excel-based macro that permits analysis of data for which tissues may have two or more scores per antibody, and permits combination of data from multiple different tissue microarrays. It accomplishes this by rendering one score per tissue per antibody from two or more scores, using one of multiple user selectable combination rules developed to account for the differing staining properties of tissue epitopes. This greatly facilitates analysis of tissue microarrays, particularly for users with large repositories of data, and may facilitate discovery of biological trends and help refine diagnostic accuracy of tissue markers in clinical samples. PMID- 16258509 TI - Epigenetic alterations in neuroendocrine tumors: methylation of RAS-association domain family 1, isoform A and p16 genes are associated with metastasis. AB - Well-differentiated neuroendocrine tumors including pancreatic endocrine tumors and carcinoid tumors are uncommon neoplasms that have site-specific differences in clinicopathological features, clinical course and genetic alterations. The epigenetic alterations in these tumors are not well characterized. We therefore compared methylation of the RAS-association domain family 1, isoform A (RASSF1A), p14, p16 and O6-methyl-guanine methyltransferase genes in neuroendocrine tumors from 47 patients including 16 pancreatic, 15 nonileal and 16 ileal neuroendocrine tumors. Methylation of the RASSF1A gene was present in 57% of tumors, p14 in 49%, p16 in 26% and O6-methyl-guanine methyltransferase in 13% of tumors. Ileal neuroendocrine tumors lacked methylation of O6-methyl-guanine methyltransferase gene (P = 0.04). RASSF1A methylation was associated with histopathologic type of tumors (P = 0.03) and lymph node metastasis (P = 0.004), and p16 methylation with older patient age (P = 0.002) and liver metastasis (P = 0.04). Two or more genes were methylated in 53% of tumors, one gene was methylated in 30% of tumors, and all four genes were unmethylated in 17% of tumors. Methylation of one or more gene was associated with older age of patients (P = 0.01), and methylation of two or more genes was associated with liver metastasis (P = 0.044). Our study shows that in neuroendocrine tumors epigenetic alterations vary by tumor subsite and clinicopathologic features, including age of onset, histopathologic type and metastasis status. PMID- 16258510 TI - p53 and c-kit (CD117) protein expression as prognostic indicators in breast phyllodes tumors: a tissue microarray study. AB - Breast phyllodes tumors are fibroepithelial neoplasms whose clinical behavior is difficult to predict on histology. There is relatively scant data on the role of biological markers. In this study, we determined if p53 and CD117 (c-kit) protein expression was predictive of behavior in a series of 335 phyllodes tumors diagnosed at the Singapore General Hospital, using immunohistochemistry on tissue microarrays. Representative areas from 250 (75%) benign, 54 (16%) borderline and 31 (9%) malignant phyllodes tumors were selected for construction of tissue microarrays using the 2 mm punch. Immunohistochemistry for p53 and CD117 was carried out using the streptavidin-biotin method. Staining proportion and intensity of both epithelial and stromal elements were analyzed. p53 immunostaining was observed in the epithelium of 28 (10%) of 278 microarrays; myoepithelium of 53 (21%) of 251 microarrays; and stromal cells in 105 (36%) of 289 microarrays. CD117 immunohistochemical reactivity was noted in epithelial and stromal components of 175 (of 267, 66%) and 17 (of 273, 6%) microarrays, respectively. Stromal p53 and CD117 protein expression was associated with tumor grade (P < 0.05). Of 43 (13%) women who suffered recurrences during the follow-up period, CD117 stromal staining predicted recurrent disease (P<0.05), but p53 was not correlative. We conclude that tissue microarrays are a convenient method for evaluating immunostaining results of large numbers of phyllodes tumors. Although positive p53 stromal immunohistochemical detection may corroborate histologic malignancy, it is CD117 protein expression in phyllodes tumor stromal cells that may be of potential utility in predicting recurrent disease. PMID- 16258511 TI - Diagnostic utility of p75 neurotrophin receptor (p75NTR) as a marker of breast myoepithelial cells. AB - We evaluated the low affinity neurotrophin receptor (p75NTR) as a marker of breast myoepithelial cells. Immunohistochemical staining for p75NTR was performed on paraffin sections of 122 malignant breast lesions, 28 benign lesions and the adjacent normal breast tissue. The staining pattern was compared to those of myosin heavy chain and p63. p75NTR immunostain was consistently positive and compatible with p63 and myosin immunoreactivity in the myoepithelial cells of the normal mammary gland, benign breast lesions (six usual ductal hyperplasias, six specimens with sclerosing adenosis, eight intraductal papillomas, six fibroadenomas), and carcinoma in situ (18 ductal carcinomas in situ, two noninvasive papillary carcinomas, two lobular carcinomas in situ). The luminal cells were negative for p75NTR, but rare positive cells were noticed in the solid areas of some of the usual ductal hyperplasias. Four of 64 invasive ductal carcinomas (6%) and all metaplastic carcinomas (n = 3, 100%) showed a variable degree of p75(NTR) positivity. No p75NTR expression was found in the malignant cells in all in situ carcinomas, invasive lobular carcinomas (n = 11), tubular carcinomas (n = 10), invasive papillary carcinomas (n = 6), mucinous carcinomas (n = 4), and medullary carcinomas (n = 2). No myosin immunoreactivity was seen in the luminal/tumor cells, but p63 pattern of staining in the luminal/tumor cells was quite similar to that of p75NTR. Although significant p75NTR immunoreactivity was noticed in the vessels, nerves, and stromal component of fibroadenomas, no difficulties in the evaluation of the immunostain of myoepithelial cells were encountered. Our study shows that p75NTR is a useful marker for breast myoepithelial cells and can be used to rule out invasive disease as well as to evaluate difficult for diagnosis sclerosing lesions. Our data suggest a role of neurotrophins in the development of fibroepithelial breast tumors and some of the breast carcinomas. PMID- 16258512 TI - Molecular diagnosis of Ewing sarcoma/primitive neuroectodermal tumor in routinely processed tissue: a comparison of two FISH strategies and RT-PCR in malignant round cell tumors. AB - Ewing sarcoma/primitive neuroectodermal tumor (EWS/PNET) is a diagnostically challenging malignant round cell tumor with signature translocations involving the EWS gene. These translocations are detectable with both reverse transcriptase polymerase chain reaction (RT-PCR) and fluorescence in situ hybridization (FISH) in formalin-fixed paraffin-embedded tissue. However, RT-PCR is less sensitive in formalin-fixed paraffin-embedded than frozen tissue. Similarly, commercial FISH probes have recently become available, but have yet to be rigorously tested in the clinical setting. Therefore, we have compared RT-PCR with FISH using 'home brew' fusion probes for Ewing sarcoma (EWS)-FLI1 and a commercial EWS break apart probe set in 67 archival round cell tumors, including 27 EWS/PNETs. Sensitivities and specificities for both FISH assays were 91 and 100%, respectively, whereas RT PCR had a sensitivity of 54% and a specificity of 85%. The break apart strategy was easier to interpret than probe fusion approach. We conclude that FISH is a more sensitive and reliable ancillary technique than RT-PCR for the diagnosis of EWS/PNET in formalin-fixed paraffin-embedded tissue, although the latter provides additional information regarding fusion transcript subtype and prognosis. The commercial break apart probe set is both readily available and easy to interpret, making it particularly attractive. Nonetheless, complex round cell tumors often benefit from molecular testing with multiple methods. PMID- 16258513 TI - Epithelioid trophoblastic tumor: clinicopathological features with an emphasis on uterine cervical involvement. AB - We report on the clinical and histological features of five cases of epithelioid trophoblastic tumor, with an emphasis on its involvement of the uterine cervix. All five patients were of reproductive age (median age 38.4 years) and all, except one, presented with vaginal bleeding 3 to 18 years after the most recent pregnancy. One patient presented with amenorrhea. Elevation of serum human chorionic gonadotropin (hCG) was seen in four cases. Pathologically, the tumor involved endocervix in three cases and involved uterine corpus in another two. All five tumors were invasive, nodular lesions consisting of epithelioid intermediate trophoblastic cells that were mononuclear with abundant eosinophilic cytoplasm, along with zones of hyaline material and necrotic debris. In three cases of cervical involvement, the neoplastic cells focally replaced endocervical surface and glandular epithelium, simulating high-grade squamous intraepithelial lesions. Immunohistochemically, all five tumors displayed focal positivity for human placental lactogen and hCG. Positive nuclear staining of p63 was seen in all five cases. All patients received total hysterectomy and various regimes of adjuvant chemotherapy. Three patients survived the tumor with no recurrences or metastases with follow-up periods of 3, 7 and 16 years. One patient is currently alive with lung metastasis 1 month after the surgery. One patient died of tumor metastasis 8 months after the diagnosis. In summary, with its unusual ability to simulate an invasive squamous cell carcinoma and other epithelioid neoplasms, epithelioid trophoblastic tumor frequently poses a diagnostic challenge, especially when involving the uterine cervix. High index of suspicion and an awareness of elevation of serum chorionic gonadotropin are crucial in reaching a correct diagnosis. PMID- 16258514 TI - MDM2 protein expression is a negative prognostic marker in breast carcinoma. AB - The protein encoded by the MDM2 oncogene inhibits the function of p53, leading to increased cell growth, avoidance of apoptosis, tolerance of genetic instability, and resistance to chemotherapy. The present study was performed to evaluate the relationship between MDM2 protein expression and survival in breast carcinoma. Two series of cases were used in this study: the first to identify the cutoff to be used in the interpretation of MDM2 immunostaining and perform preliminary survival analysis, and a second, independent series, to validate the findings from the first series and to perform multivariate analysis. For both series, archival sections of tissue microarrays were stained with anti-MDM2 antibody (NeoMarkers, Fremont, CA, USA) and MDM2 staining intensity was scored semiquantitatively. In the first series, 49 of 362 (14%) interpretable cases were positive for MDM2 expression, with 35 (10%) showing weak positivity and 14 (4%) strong positivity. Patients with MDM2-positive tumours had a significantly worse disease-specific survival than patients with MDM2-negative tumours (P=0.0022, 10 year DSS 61% (95% CI: 45-73) vs 73% (95% CI: 67-77)). No significant difference in survival was observed between patients with strongly and weakly MDM2-positive tumours (P=0.3). Accordingly, in the independent validation series weak and strong MDM2 positivity were combined and considered to be MDM2 positive. MDM2 expression was seen in 230/1747 (13%) interpretable cases in this series, with a significant difference (P<0.0001) in DSS between MDM2-negative and MDM2-positive cases (10 year DSS 58% (95% CI: 51-64) vs 73% (95% CI: 70-75)). MDM2 was an independent prognostic marker (HR=1.35, P=0.02) in a Cox regression model including MDM2 expression, tumour grade, nodal status, ER status and tumour size. Immunohistochemical studies of MDM2 in more than 2000 breast carcinomas show that MDM2 is an independent negative prognostic marker. PMID- 16258515 TI - KIT is highly expressed in adenoid cystic carcinoma of the breast, a basal-like carcinoma associated with a favorable outcome. AB - Recent biological studies have classified breast carcinomas into HER2 overexpressing, estrogen receptor-positive/luminal, basal- and normal-like groups. According to this new biological classification, the objectives of our study were to assess the clinical, morphologic and immunophenotypic characteristics of adenoid cystic carcinoma of the breast in order to classify this subtype of breast carcinoma. A total of 18 cases of adenoid cystic carcinoma were identified from the Institut Curie files. Clinical information was available for 16 patients with a median follow-up of 6.5 years. Morphologically, all tumors were graded according to the system defined by Kleer and Oberman (histologic and nuclear grade). Immunophenotype was assessed with anti-ER, PR, HER-2, KIT, basal (CK5/6) and luminal cytokeratins (CK8/18) and p63 antibodies. One out of 18 tumors was nuclear grade 1 (16%), nine were nuclear grade 2 (50%) and eight were nuclear grade 3 (44%). All cases were estrogen receptor, progesterone receptor and HER-2 negative. Epithelial cells were strongly positive around glandular lumina with one or both cytokeratins, identifying the coexistence of CK5/6+ cells, CK5/6 and CK8/18+ cells, CK8/18+ cells and p63+ cells. All cases (100%) were also KIT positive. In all, 15 patients were treated by surgery. Nine of them received adjuvant radiotherapy. Follow-up was available for 16 patients. In all, 14 patients were alive. Two of them, initially treated by surgery only, presented a local recurrence. Two patients died (one of them treated by radiation therapy only died from her disease). Our study shows that adenoid cystic carcinoma of the breast is a special, estrogen receptor, progesterone receptor, HER-2 negative and highly KIT-positive, basal-like breast carcinoma, associated with an excellent prognosis. This highly specific immunophenotype could be useful to differentiate adenoid cystic carcinoma of the breast from other subtypes of breast carcinoma such as cribriform carcinoma. PMID- 16258516 TI - Gastrointestinal beta2microglobulin amyloidosis in hemodialysis patients: biochemical analysis of amyloid proteins in small formalin-fixed paraffin embedded tissue specimens. AB - We present here a first report on the biochemical analysis of intestinal amyloid deposits found in two cases of hemodialysis-related amyloidosis. A new microtechnique was applied for extraction and immunochemical/chemical characterization of amyloid proteins in small amounts of fixed tissue, thus allowing precise identification of beta2microglobulin amyloid (Abeta2M) in both cases studied. The molecular mass of the identified amyloid beta2M was close to that of intact beta2M (12 kDa), with no evidence of the products of proteolytic fragmentation of these molecules. The isoelectrofocusing of the purified Abeta2M demonstrated a shift to more acidic pI as compared to the normal beta2M analyzed under the same experimental conditions. The obtained data suggest that the intestinal amyloid deposits in dialysis-related amyloidosis contain disease specific beta2M isoforms, which could play a role in the pathogenesis of amyloid disease. The new methodology used might be useful in obtaining precise diagnosis of amyloidosis that is necessary for appropriate therapy, and also provide new important information on the chemical structure of amyloid proteins. PMID- 16258517 TI - Mast cells and cutaneous malignancies. AB - This paper reviews the role of mast cells in the development and progression of basal cell carcinoma, squamous cell carcinoma and malignant melanoma. Mast cells accumulate around cutaneous malignancies. Current evidence suggests that mast cells contribute to the tumorigenesis of cutaneous malignancies through four mechanisms. (1) Immunosuppression: Ultraviolet-B radiation, the most important initiator of cutaneous malignancies, activates mast cells. Upon irradiation of the skin, trans-urocanic acid in the epidermis isomerizes to cis-urocanic acid, which stimulates neuropeptide release from neural c-fibers. These neuropeptides in turn trigger histamine secretion from mast cells, leading to suppression of the cellular immune system. (2) Angiogenesis: Mast cells are the major source of vascular endothelial growth factor in basal cell carcinoma and malignant melanoma. Vascular endothelial growth factor is one of the most potent angiogenic factors, which also induces leakage of other angiogenic factors across the endothelial cell wall into the matrix. Mast cell proteases reorganize the stroma to facilitate endothelial cell migration. As well, heparin, the dominant mast cell proteoglycan, assists in blood-borne metastasis. (3) Degradation of extracellular matrix: Through its own proteases, and indirectly via interaction with other cells, mast cells participate in degradation of the matrix, which is required for tumor spread. (4) Mitogenesis: Mast cell mediators including fibroblast growth factor-2 and interleukin-8 are mitogenic to melanoma cells. Current evidence supports an accessory role for mast cells in the development and progression of cutaneous malignancies. Emerging data, however, also suggest that mast cells might, in fact, have opposing roles in tumor biology, and the microenvironment could polarize mast cells to possess either promoting or inhibitory effects on tumors. PMID- 16258518 TI - Chromogenic in situ hybridization-detected hotspot MYCN amplification associates with Ki-67 expression and inversely with nestin expression in neuroblastomas. AB - Since neuroblastomas are intratumorally heterogeneous, the analysis of genetic and biologic features of randomly selected tumor specimen spots may lead to erroneous conclusions. Our purpose was therefore to construct an easily assessable and strictly defined strategy to unify the detection of various molecular markers in paraffin-embedded neuroblastoma samples. We selected tumor specimen spots of highest proliferation activity, that is, hotspots, for the analysis of MYCN amplification status and proliferation-associated molecular markers, such as nestin, which role in neuroblastoma specimens was evaluated for the first time. Using a chromogenic in situ hybridization (CISH) technique, we showed that patients with a MYCN copy number higher than six in anti-Ki-67 detected hotspots have significantly worse overall survival prognosis than patients with low MYCN copy numbers (P = 0.0006). The chosen cutoff value of six was shown to dichotomize MYCN-amplified neuroblastomas at least as specifically as Southern blot hybridization, in which amplification was defined by a copy number of > or = 10. Interestingly, we also detected without difficulty MYCN amplified neuroblastic cells in bone marrow samples using the CISH technique. The proliferation activity, assessed with an anti-Ki-67-based proliferation index, was significantly higher in MYCN-amplified than in nonamplified hotspots. The proliferation indices of the hotspots had also a significant correlation with the prognosis (International Classification) and histological type, whereas the proliferation accelerator Id2 did not associate with any of the mentioned parameters. The expression of nestin associated inversely with MYCN amplification (P = 0.018), which challenges a previously suggested role of nestin in neuroblastomas. In summary, hotspot focusing provides a means of analyzing proliferation-associated markers in neuroblastomas, and together with the CISH detection of the MYCN copy number enables an easy and reliable examination of MYCN status in neuroblastomas. PMID- 16258519 TI - Efficient differentiation of human embryonic stem cells to definitive endoderm. AB - The potential of human embryonic stem (hES) cells to differentiate into cell types of a variety of organs has generated much excitement over the possible use of hES cells in therapeutic applications. Of great interest are organs derived from definitive endoderm, such as the pancreas. We have focused on directing hES cells to the definitive endoderm lineage as this step is a prerequisite for efficient differentiation to mature endoderm derivatives. Differentiation of hES cells in the presence of activin A and low serum produced cultures consisting of up to 80% definitive endoderm cells. This population was further enriched to near homogeneity using the cell-surface receptor CXCR4. The process of definitive endoderm formation in differentiating hES cell cultures includes an apparent epithelial-to-mesenchymal transition and a dynamic gene expression profile that are reminiscent of vertebrate gastrulation. These findings may facilitate the use of hES cells for therapeutic purposes and as in vitro models of development. PMID- 16258520 TI - 3D-confocal structural analysis of bone marrow-derived renal tubular cells during renal ischemia/reperfusion injury. AB - Bone marrow cells (BMC) have been shown to migrate into injured sites for parenchymal repair. However, the extent of BMC involvement is controversial. To determine whether and to what extent BMC contribute to renal parenchymal repair, we employed three-dimensional (3D) fluorescent confocal microscopy/video in renal warm and cold ischemia/reperfusion (I/R) injury using enhanced green fluorescent protein transgenic rats and their radiation chimeras. After induction of renal warm I/R injury in chimeras, BM-derived renal tubular cells were found in 2D microscopy as isolated single cells or clusters of 2-3 cells. Likewise, cold I/R injury resulted in host-derived tubular cells with frequencies approximately 0.2%. However, stringent confocal microscopic analysis and 3D image construction revealed that BM-derived tubules identified in 2D images were frequently artifacts of overlapping cells separately stained with different markers. The actual frequency in 3D analysis was approximately one-fourth of that seen in 2D analysis. 3D confocal imaging precisely detected BM-derived tubular epithelial cells and could be useful to study BMC contribution to tissue repair. PMID- 16258521 TI - GISTs with PDGFRA exon 14 mutations represent subset of clinically favorable gastric tumors with epithelioid morphology. AB - Gastrointestinal stromal tumors (GISTs) are common mesenchymal tumors of the gastrointestinal tract. Activating KIT or PDGFRA (platelet-derived growth factor receptor alpha) mutations have been shown to be a major force in GIST pathogenesis. Recently, a previously undescribed N659K PDGFRA exon 14 mutation has been reported in GISTs. The purpose of this study was to evaluate the frequency of GISTs with PDGFRA exon 14 mutations and define the clinicopathologic profile of such tumors. In all, 200 GISTs negative for mutations in KIT exons 9, 11, 13 and 17 and PDGFRA exons 12 and 18 were evaluated for PDGFRA exon 14 mutations by PCR amplification and direct sequencing. Mutations were found in 11 of 119 (9%) gastric GISTs. None of the 81 GISTs from other than gastric location had such a PDGFRA mutation. A majority of these mutations (eight cases) represented simple 2125C>A or C>G missense mutations, leading to substitution of the lysine for asparagine (N659K). However, in two cases, 2123A>T missense mutations leading to substitution of the tyrosine for asparagine (N659Y) was found instead. Of 11 PDGFRA N659-mutant GISTs, 10 had pure epithelioid morphology. One tumor had mixed, predominantly spindle and focally epithelioid cell morphology. Frequency of PDGFRA N659-mutant GISTs among pure epithelioid GISTs was almost 19%. Immunohistochemically, the majority (64%) of these tumors lacked KIT expression or showed only focal scattered KIT positivity. Tumor size ranged from 2.5 to 16 cm (average 7.1 cm). Low mitotic activity, 5 cm tumors. Based on mitotic activity and tumor size, six tumors were classified as probably benign with very low malignant potential. Low to moderate malignant potential and high malignant potential was suggested in three and two tumors, respectively. In four cases with moderate or high malignant potential GISTs, a long-term follow-up (average 235.5 months) showed favorable course of disease. PMID- 16258522 TI - Impaired myocardial capillarogenesis and increased adaptive capillary growth in FGF2-deficient mice. AB - Basic fibroblast growth factor (FGF2) plays a major role in angiogenesis and capillary growth. In contrast to vascular endothelial growth factor, which is required for proliferation and survival of endothelial cells, FGF2 does not seem to be essential since the Fgf2 knockout is not lethal. Therefore, the precise genetic and physiological roles of FGF2 for capillary development and adaptation remain to be determined. Here we show that myocardial capillary supply is normal at birth, but significantly reduced by approximately 25% in adult Fgf2+/- and Fgf2-/- mice as compared with wild-type littermates. In contrast, after induction of myocardial hypertrophy by continuous infusion of angiotensin II (ANG II) for 6 days marked capillary growth was seen in both Fgf2+/- and Fgf2-/- mice, but not in wild-type littermates. These data demonstrate that two intact Fgf2 genes are necessary for normal capillary development after birth, whereas FGF2 seems to be dispensable for adaptive myocardial capillary growth in the adult mouse. PMID- 16258523 TI - Amplifications of the epidermal growth factor receptor gene (egfr) are common in phyllodes tumors of the breast and are associated with tumor progression. AB - Phyllodes tumors of the breast are rare biphasic tumors with the potential for invasion and metastatic spread. An important role of the epidermal growth factor receptor (EGFR) in phyllodes tumors has been proposed. However, detailed pathogenetic mechanisms remained unclear. We investigated 58 phyllodes tumors of the breast (40 benign, 10 borderline and eight malignant) by means of egfr fluorescence in situ hybridization (FISH) and gene dosage PCR for a regulatory sequence within intron 1 of egfr. Immunohistochemical staining was performed for EGFR, p16, p21, p27, p53, c-myc, Cyclin A, Cyclin D1, Cyclin E, c-kit and Ki67. Immunopositivity for EGFR was detected in 19% of phyllodes tumors (75% of all malignant tumors) in stromal tumor cells but not in the epithelial component. Whole-gene amplifications were seen by FISH in 15.8% (in stromal cells only) and intron 1 amplifications by gene dosage PCR in as much as 41.8% of all phyllodes tumors. Significant correlations were seen between tumor grade on the one hand and EGFR overexpression (P=0.001) and intron 1 amplifications (P<0.05) on the other. EGFR overexpression further correlated positively with immunohistochemical staining for p53, p16, Cyclin A, Cyclin E, Ki67 and c-kit. Presence of intron 1 amplifications correlated with p16 (P<0.01), p21 (P=0.009) and p53 immunoreactivity (P<0.001). Neither EGFR overexpression nor whole-gene amplification was observed in a control series of 167 fibroadenomas and only one of 43 (2.3%) exhibited intron 1 amplification in gene dosage PCR. In conclusion, our results show for the first time that activating mutations in and overexpression of egfr are associated with the progression in grade of phyllodes tumors of the breast. The observed association between intron 1 amplification and overexpression of EGFR provides further insight into regulation mechanisms of EGFR overexpression. PMID- 16258524 TI - AM 251 produces sustained reductions in food intake and body weight that are resistant to tolerance and conditioned taste aversion. AB - The cannabinoid 1 (CB(1)) receptor has been implicated in the regulation of food intake. Here, we examine the effect of the CB(1) receptor antagonist AM 251 on food intake and body weight over a prolonged period. Further, we examine whether AM 251 produces conditioned taste aversion (CTA) and if sustained antagonism at central receptors contributes to its anorectic effect. The effect of AM 251 of food intake and body weight was examined in daily (1 mg kg(-1)) and 5-day (5 mg kg(-1)) dosing schedules. Matching reductions in food intake and body weight were observed in both paradigms. A single administration of AM 251 (5 mg kg(-1)) significantly reduced food intake for 4 days. Tolerance to the anorectic effects of AM 251 did not develop in either dosing strategy. Active avoidance of AM 251 (3; 5 mg kg(-1), i.p.) was examined using a CTA assay. Rats showed no evidence of CTA associated with AM 251. We investigated the sustained effect of AM 251 (5 mg kg(-1), i.p.) on CB(1) receptors in the hypothalamus using Delta(9) tetrahydrocannabinol (8 mg kg(-1), i.p.) induced hypothermia. AM 251 initially blocked hypothermia, but this effect was not seen 2 or 4 days later. The results demonstrate that smaller, or infrequent, administrations of AM 251 can produce sustained reductions in food intake and body weight in rat. Reductions in food intake were sustained longer than AM 251 antagonized the effects of a CB(1) receptor agonist in the hypothalamus, and occurred independently of CTA. PMID- 16258525 TI - A delayed ATP-elicited K+ current in freshly isolated smooth muscle cells from mouse aorta. AB - Adenosine 5'-triphosphate (ATP) activated two sequential responses in freshly isolated mouse aortic smooth muscle cells. In the first phase, ATP activated Ca(2+)-dependent K(+) or Cl(-) currents and the second phase was the activation of a delayed outward current with a reversal potential of -75.9 +/- 1.4 mV. A high concentration of extracellular K(+) (130 mM) shifted the reversal potential of the delayed ATP-elicited current to -3.5 +/- 1.3 mV. The known K(+)-channel blockers, iberiotoxin, charybdotoxin, glibenclamide, apamin, 4-aminopyridine, Ba(2+) and tetraethylammonium chloride all failed to inhibit the delayed ATP elicited K(+) current. Removal of ATP did not decrease the amplitude of the ATP elicited current back to the control values. The simultaneous recording of cytosolic free Ca(2+) and membrane currents revealed that the first phase of the ATP-elicited response is associated with an increase in intracellular Ca(2+), while the second delayed phase develops after the return of cytosolic free Ca(2+) to control levels.ATP did not activate Ca(2+)-dependent K(+) currents, but did elicit Ca(2+)-independent K(+) currents, in cells dialyzed with ethylene glycol bis (2-aminoethylether)-N,N,N',N'-tetraacetic acid (EGTA). The delay of activation of Ca(2+)-independent currents decreased from 10.5 + 3.4 to 1.27 +/- 0.33 min in the cells dialyzed with 2 mM EGTA. Adenosine alone failed to elicit a Ca(2+)-independent K(+) current but simultaneous application of ATP and adenosine activated the delayed K(+) current. Intracellular dialysis of cells with guanosine 5'-O-(2-thiodiphosphate) transformed the Ca(2+)-independent ATP elicited response from a sustained to a transient one. A phospholipase C inhibitor, U73122 (1 microM), was shown to abolish the delayed ATP-elicited response. These results indicate that the second phase of the ATP-elicited response was a delayed Ca(2+)-independent K(+) current activated by exogenous ATP. This phase might represent a new vasoregulatory pathway in vascular smooth muscle cells. PMID- 16258527 TI - PDE5 inhibition and fibrosis. AB - Can PDE5 inhibitors inhibit fibrosis? Is this a direct effect of PDE5 inhibitors on the fibrotic cascade? If this hypothesis is found true, then an entire new algorithm for treatment of ED and other diseases may be realized. Three recent manuscripts this year discuss this question. Dr Jackie Corbin, a renowned PDE5 expert, offers a perspective in light of its potential clinical relevance. PMID- 16258526 TI - Acute denervation alters the epithelial response to adrenoceptor activation through an increase in alpha1-adrenoceptor expression on villus enterocytes. AB - Loss of sympathetic input due to intestinal denervation results in hypersensitivity and increased intestinal secretion. It is unknown whether denervation-induced alterations in intestinal epithelial physiology are the result of changes in adrenoceptors on enterocytes (ENTs). The purpose of this study was to examine adrenoceptor distribution and pharmacology on small intestinal ENTs following acute intestinal denervation. Lewis rats underwent small bowel transplantation (SBT) or sham operation and proximal small intestinal segments were harvested 1, 2 and 4 weeks postoperatively. Intestinal electrolyte movement was assessed using short-circuit current (Isc) measurements of stripped epithelial sheets following stimulation with phenylephrine (PE), an alpha(1) adrenoceptor agonist. The presence of adrenoceptor subtypes on separated villus and crypt ENTs was assessed using flow cytometry. Alpha(1)-adrenoceptors were found on approximately 27% of jejunal villus ENTs, but not crypt ENTs, following acute extrinsic denervation. ENTs from the Lewis rat have few beta-adrenoceptors. Alpha(1)-adrenoceptor stimulation of acutely denervated intestinal epithelial sheets decreased Isc by -13.45%. This effect was mediated by a reduction in chloride (Cl(-)) secretion; the absence of Cl(-) reversed the Isc to +13.79%. In conclusion, loss of sympathetic innervation to the gastrointestinal epithelium causes acute upregulation of alpha(1)-adrenoceptors on villus ENTs, leading to inhibition of Cl(-) secretion at the villus tip. The increase in adrenoceptors may reflect a compensatory mechanism to combat the increased secretory state of the bowel due to the loss of the sympathetic innervation and tonic control over intestinal secretion. PMID- 16258529 TI - Kinetics of dendritic cell chimerism and T cell chimerism in allogeneic hematopoietic stem cell recipients. AB - Dendritic cells (DC) as potent antigen-presenting cells (APC) and T cells as effector cells play an essential role in the pathophysiology of both graft-versus host (GvH) and graft-versus-leukemia (GvL) reactions after transplantation. Therefore, we determined the kinetics of DC and T-cell chimerism establishment after allogeneic hematopoietic cell transplantation (AHCT) in a group of 144 patients, using fluorescence-activated cell sorting (FACS) or magnetic cell sorting (MACS) followed by FISH or STR-PCR analysis for chimerism evaluation. In all, three cell lines investigated (CD3(+) T cells, CD11c(+) DC1 and CD123(+) DC2), we found a rapid and consistent establishment of complete donor chimerism (CDC) in over 70% of all patients during the first 6 weeks after AHCT. The rate of patients with CDC increased significantly over time within the first year after transplantation. A related donor (P=0.004) as well as an underlying lymphatic leukemia (P=0.03) were found to be significantly associated with development of MC in T cells. No significant correlation between DC or T cell chimerism and GvHD or relapse was detected. Our results thus demonstrate a fast and stable CDC in DC1, DC2 and T cells after AHCT that continuously increases over time in nearly all patients. PMID- 16258530 TI - A successful case of allogeneic bone marrow transplantation for osteosarcoma with multiple metastases of lung and bone. PMID- 16258532 TI - Allogeneic stem cell transplantation from unrelated donor for class 3 beta thalassemia major using reduced-intensity conditioning regimen. PMID- 16258531 TI - Allogeneic stem-cell transplantation in patients with refractory acute leukemia: a long-term follow-up. AB - We examined retrospectively 44 patients with refractory acute leukemia (acute myeloid leukemia (AML)/acute lymphoblastic leukemia=25/19) who underwent allogeneic transplantation at our center between 11/1990 and 04/2004. The median leukemic blasts was 25% and age 28 years (range, 3-56). Twenty-one patients had untreated relapse, 13 failed reinduction, eight in partial remission and two aplastic. Conditioning was myeloablative using cyclophosphamide, busulfan, total body irradiation and etoposide (Bu/Cy/VP, n=22; TBI/Cy/VP, n=17; others, n=5) followed by marrow or peripheral blood transplant (n=23/21) from unrelated or related donors (n=28/16). All patients had graft-versus-host disease (GVHD) prophylaxis with cyclosporin and methotrexate. One patient experienced late graft failure. Severe acute-GVHD and chronic-GVHD appeared in eight and 14 patients, respectively. Thirteen patients (30%) remain alive after a median of 25.3 months (range, 2.4-134.1); with 31 deaths, mostly from relapse (n=15) and infections (n=12). Overall survival (OS) and progression-free survival (PFS) at 5 years was 28 and 26%, respectively. OS and PFS were significantly better with blasts < or =20% and time to transplant < or =1 year while transplant-related mortality was less with the use of TBI. We conclude that patients with refractory leukemia can benefit from allogeneic BMT, especially with < or =20% marrow blast. PMID- 16258533 TI - Prognostic significance of platelet recovery pattern after allogeneic HLA identical sibling transplantation and its association with severe acute GVHD. AB - Thrombocytopenia (TP) is a frequent complication after allogeneic stem cell transplantation (SCT) and regarded as a poor prognostic factor when assessed beyond day 100. However, little is known about the clinical significance of the platelet recovery pattern before chronic graft-versus-host disease (GVHD) develops. Eighty-five patients undergoing HLA-identical sibling SCT were stratified according to their platelet recovery pattern between day +30 and +90 and the transplant outcomes analyzed, along with the association of each component of the acute GVHD grading system. Fifteen patients (18%) were classified with persistent TP, 33 patients (39%) with unstable TP, and 37 patients (43%) as non-TP. Persistent TP, which was strongly associated with severe acute GVHD (P<0.001), exhibited the worst 2-year OS (P<0.0001) and highest NRM (P<0.0001) and opportunistic infection rates (P<0.0001). In multivariate analyses, the platelet recovery pattern was identified as an independent prognostic factor (P=0.02) together with the disease risk (P=0.02) in terms of OS, and the only independent prognostic factor in terms of NRM (P=0.005) and the incidence of infectious events (P<0.001). Persistent TP was strongly associated with the development of extensive chronic GVHD (P=0.03). The platelet recovery pattern between day +30 and +90 can be used to predict the prognosis of SCT recipients. PMID- 16258534 TI - Cure of multiple myeloma -- more hype, less reality. AB - Randomized studies have firmly established the role of autologous transplant as initial therapy in multiple myeloma (MM). Indeed, MM has emerged as the commonest indication for autologous SCT in North America. The conceptual basis for high dose therapy is the goal of complete remission (CR) through steep reduction in tumor burden affected by single and tandem transplants. Careful analysis of the data challenges the notion of CR as a surrogate to success. Intrinsically aggressive MM, defined by known unfavorable biologic risk factors, overrides the benefit of CR. In contrast, subgroups of patients with favorable biological risk factors may achieve prolonged survival, often without ever achieving CR. Unfortunately, even with tandem transplants, there is no plateau in survival curves. To this end, sequential autologous followed by nonmyeloablative allotransplants are a novel attempt at 'curing' myeloma, but the results thus far have failed to show a definite plateau in survival. Given the improvements in supportive care and concomitant reduction in transplant-related mortality, conventional myeloablative allogeneic transplants need to be re-examined as an option in high-risk aggressive myeloma. At the same time, novel antimyeloma therapies, newer risk stratification and staging tools are transforming the treatment algorithm. We examine the changing role of transplantation in myeloma in the context of novel drug therapy, biologic risk stratification and improving supportive care while arguing that the current 'one size fits all' transplant approaches are far from a cure. PMID- 16258535 TI - Silencing of microRNAs in vivo with 'antagomirs'. AB - MicroRNAs (miRNAs) are an abundant class of non-coding RNAs that are believed to be important in many biological processes through regulation of gene expression. The precise molecular function of miRNAs in mammals is largely unknown and a better understanding will require loss-of-function studies in vivo. Here we show that a novel class of chemically engineered oligonucleotides, termed 'antagomirs', are efficient and specific silencers of endogenous miRNAs in mice. Intravenous administration of antagomirs against miR-16, miR-122, miR-192 and miR 194 resulted in a marked reduction of corresponding miRNA levels in liver, lung, kidney, heart, intestine, fat, skin, bone marrow, muscle, ovaries and adrenals. The silencing of endogenous miRNAs by this novel method is specific, efficient and long-lasting. The biological significance of silencing miRNAs with the use of antagomirs was studied for miR-122, an abundant liver-specific miRNA. Gene expression and bioinformatic analysis of messenger RNA from antagomir-treated animals revealed that the 3' untranslated regions of upregulated genes are strongly enriched in miR-122 recognition motifs, whereas downregulated genes are depleted in these motifs. Analysis of the functional annotation of downregulated genes specifically predicted that cholesterol biosynthesis genes would be affected by miR-122, and plasma cholesterol measurements showed reduced levels in antagomir-122-treated mice. Our findings show that antagomirs are powerful tools to silence specific miRNAs in vivo and may represent a therapeutic strategy for silencing miRNAs in disease. PMID- 16258536 TI - Protection of macaques from vaginal SHIV challenge by vaginally delivered inhibitors of virus-cell fusion. AB - Human immunodeficiency virus type 1 (HIV-1) continues to spread, principally by heterosexual sex, but no vaccine is available. Hence, alternative prevention methods are needed to supplement educational and behavioural-modification programmes. One such approach is a vaginal microbicide: the application of inhibitory compounds before intercourse. Here, we have evaluated the microbicide concept using the rhesus macaque 'high dose' vaginal transmission model with a CCR5-receptor-using simian-human immunodeficiency virus (SHIV-162P3) and three compounds that inhibit different stages of the virus-cell attachment and entry process. These compounds are BMS-378806, a small molecule that binds the viral gp120 glycoprotein and prevents its attachment to the CD4 and CCR5 receptors, CMPD167, a small molecule that binds to CCR5 to inhibit gp120 association, and C52L, a bacterially expressed peptide inhibitor of gp41-mediated fusion. In vitro, all three compounds inhibit infection of T cells and cervical tissue explants, and C52L acts synergistically with CMPD167 or BMS-378806 to inhibit infection of cell lines. In vivo, significant protection was achieved using each compound alone and in combinations. CMPD167 and BMS-378806 were protective even when applied 6 h before challenge. PMID- 16258537 TI - Signal amplification in molecular imaging by pretargeting a multivalent, bispecific antibody. AB - Here we describe molecular imaging of cancer using signal amplification of a radiotracer in situ by pretargeting a multivalent, bispecific antibody to carcinoembryonic antigen (CEA), which subsequently also captures a radioactive hapten-peptide. Human colon cancer xenografts as small as approximately 0.15 g were disclosed in nude mice within 1 h of giving the radiotracer, with tumor/blood ratios increased by >or=40-fold (approximately 10:1 at 1 h, approximately 100:1 at 24 h), compared to a (99m)Tc-labeled CEA-specific F(ab') used clinically for colorectal cancer detection, while also increasing tumor uptake tenfold ( approximately 20% injected dose/g) under optimal conditions. This technology could be adapted to other antibodies and imaging modalities. PMID- 16258538 TI - Platelets mediate cytotoxic T lymphocyte-induced liver damage. AB - We found that platelet depletion reduces intrahepatic accumulation of virus specific cytotoxic T lymphocytes (CTLs) and organ damage in mouse models of acute viral hepatitis. Transfusion of normal but not activation-blocked platelets in platelet-depleted mice restored accumulation of CTLs and severity of disease. In contrast, anticoagulant treatment that prevented intrahepatic fibrin deposition without reducing platelet counts did not avert liver injury. Thus, activated platelets contribute to CTL-mediated liver immunopathology independently of procoagulant function. PMID- 16258540 TI - SMC1beta-deficient female mice provide evidence that cohesins are a missing link in age-related nondisjunction. AB - Mitotic chromosome segregation is facilitated by the cohesin complex, which maintains physical connections between sister chromatids until anaphase. Meiotic cell division is considerably more complex, as cohesion must be released sequentially to facilitate orderly segregation of chromosomes at both meiosis I and meiosis II. This necessitates meiosis-specific cohesin components; recent studies in rodents suggest that these influence chromosome behavior during both cell division and meiotic prophase. To elucidate the role of the meiosis-specific cohesin SMC1beta (encoded by Smc1l2) in oogenesis, we carried out meiotic studies of female SMC1beta-deficient mice. Our results provide the first direct evidence that SMC1beta acts as a chiasma binder in mammals, stabilizing sites of exchange until anaphase. Additionally, our observations support the hypothesis that deficient cohesion is an underlying cause of human age-related aneuploidy. PMID- 16258541 TI - Inherited susceptibility to lung cancer may be associated with the T790M drug resistance mutation in EGFR. AB - Somatic activating mutations in EGFR identify a subset of non-small cell lung cancer that respond to tyrosine kinase inhibitors. Acquisition of drug resistance is linked to a specific secondary somatic mutation, EGFR T790M. Here we describe a family with multiple cases of non-small cell lung cancer associated with germline transmission of this mutation. Four of six tumors analyzed showed a secondary somatic activating EGFR mutation, arising in cis with the germline EGFR mutation T790M. These observations implicate altered EGFR signaling in genetic susceptibility to lung cancer. PMID- 16258542 TI - An evaluation of HapMap sample size and tagging SNP performance in large-scale empirical and simulated data sets. AB - A substantial investment has been made in the generation of large public resources designed to enable the identification of tag SNP sets, but data establishing the adequacy of the sample sizes used are limited. Using large-scale empirical and simulated data sets, we found that the sample sizes used in the HapMap project are sufficient to capture common variation, but that performance declines substantially for variants with minor allele frequencies of <5%. PMID- 16258543 TI - Sex-specific role of Drosophila melanogaster HP1 in regulating chromatin structure and gene transcription. AB - Drosophila melanogaster heterochromatin protein 1 (HP1a or HP1) is believed to be involved in active transcription, transcriptional gene silencing and the formation of heterochromatin. But little is known about the function of HP1 during development. Using a Gal4-induced RNA interference system, we showed that conditional depletion of HP1 in transgenic flies resulted in preferential lethality in male flies. Cytological analysis of mitotic chromosomes showed that HP1 depletion caused sex-biased chromosomal defects, including telomere fusions. The global levels of specific histone modifications, particularly the hallmarks of active chromatin, were preferentially increased in males as well. Expression analysis showed that approximately twice as many genes were specifically regulated by HP1 in males than in females. Furthermore, HP1-regulated genes showed greater enrichment for HP1 binding in males. Taken together, these results indicate that HP1 modulates chromosomal integrity, histone modifications and transcription in a sex-specific manner. PMID- 16258544 TI - Magnetic resonance tracking of dendritic cells in melanoma patients for monitoring of cellular therapy. AB - The success of cellular therapies will depend in part on accurate delivery of cells to target organs. In dendritic cell therapy, in particular, delivery and subsequent migration of cells to regional lymph nodes is essential for effective stimulation of the immune system. We show here that in vivo magnetic resonance tracking of magnetically labeled cells is feasible in humans for detecting very low numbers of dendritic cells in conjunction with detailed anatomical information. Autologous dendritic cells were labeled with a clinical superparamagnetic iron oxide formulation or (111)In-oxine and were co-injected intranodally in melanoma patients under ultrasound guidance. In contrast to scintigraphic imaging, magnetic resonance imaging (MRI) allowed assessment of the accuracy of dendritic cell delivery and of inter- and intra-nodal cell migration patterns. MRI cell tracking using iron oxides appears clinically safe and well suited to monitor cellular therapy in humans. PMID- 16258545 TI - Design of siRNAs producing unstructured guide-RNAs results in improved RNA interference efficiency. AB - In RNA interference (RNAi), guide RNAs direct RNA-induced silencing complexes (RISC) to their mRNA targets, thus enabling the cleavage that leads to gene silencing. We describe a strong inverse correlation between the degree of guide RNA secondary structure formation and gene silencing by small interfering (si)RNA. Unstructured guide strands mediate the strongest silencing whereas structures with base-paired ends are inactive. Thus, the availability of terminal nucleotides within guide structures determines the strength of silencing. A to G and C to U base exchanges, which involve wobble base-pairing with the target but preserve complementarity, turned inactive into active guide structures, thereby expanding the space of functional siRNAs. Previously observed base degenerations among mature micro (mi)RNAs together with the data presented here suggest a crucial role of the guide-RNA structures in miRNA action. The analysis of the effect of the secondary structures of guide-RNA sequences on RNAi efficiency provides a basis for better understanding RNA silencing pathways and improving the design of siRNAs. PMID- 16258546 TI - Endocytic control of epithelial polarity and proliferation in Drosophila. AB - Intracellular protein transport is a key factor in epithelial cell polarity. Here we report that mutations in two core components of the vesicle trafficking machinery - a syntaxin and a Rab protein - cause an expansion of the apical membrane domain of Drosophila melanogaster epithelia; this polarity defect is coupled with overproliferation to form neoplastic tumours. Surprisingly, these proteins are associated with the endocytic, and not the exocytic, pathway. The syntaxin Avalanche (Avl) localizes to early endosomes, and loss of avl results in the cellular accumulation of specific membrane proteins, including the Notch signalling receptor and the polarity determinant Crumbs (Crb). Protein accumulation results from a failure in endosomal entry and progression towards lysosomal degradation; these and other avl phenotypes are also detected in Rab5 null mutant cells. Overexpression of Crb alone is sufficient to induce overproliferation of wild-type imaginal tissue, suggesting that polarity alterations in avl and Rab5 mutants directly contribute to tumour formation. Our findings reveal a critical and specific role for endocytic traffic in the control of both apico-basal polarity and cell proliferation. PMID- 16258547 TI - Cardiovascular disease in the elderly: a challenge for science and clinical care. PMID- 16258548 TI - How and when will cardiac xenotransplantation enter the clinic? The recurrent debate has gained in realism. PMID- 16258563 TI - Does combined clopidogrel and statin therapy benefit patients with non-ST elevation myocardial infarction? PMID- 16258564 TI - Can multislice CT detect coronary artery disease accurately? PMID- 16258565 TI - Open versus endovascular abdominal aortic aneurysm repair: which offers the best long-term outcome? PMID- 16258566 TI - Do patients with acute vertebrobasilar occlusion benefit from an aggressive treatment strategy? PMID- 16258567 TI - Are paclitaxel-eluting stents safe and effective in patients with acute coronary syndromes? PMID- 16258568 TI - Realizing the clinical potential of ischemic preconditioning and postconditioning. AB - After an acute myocardial infarction (AMI), early reperfusion by thrombolysis or primary percutaneous coronary intervention remains the most-effective strategy for limiting the size of an evolving infarct. The mortality from AMI, however, remains significant, due partly to the lethal reperfusion injury that occurs on reperfusing the ischemic myocardium. Novel cardioprotective strategies are required to target this form of injury. In ischemic preconditioning transient, nonlethal episodes of myocardial ischemia and reperfusion before the index ischemic episode reduce infarct size. The cardioprotective potential of ischemic preconditioning has not been realized in clinical practice because it necessitates an intervention applied before the onset of AMI, which is difficult to predict. A more-amenable approach to cardioprotection is to intervene at the onset of reperfusion, the timing of which is under the control of the operator. In this regard, ischemic postconditioning, in which transient episodes of myocardial ischemia and reperfusion administered at the onset of reperfusion reduce infarct size, constitutes one such intervention. Interestingly, studies suggest that ischemic preconditioning and postconditioning activate the same signaling pathway at the time of reperfusion, thereby offering a common target for cardioprotection. Therefore, the pharmacologic recruitment of this signaling pathway at the time of myocardial reperfusion might allow one to harness the cardioprotective potential of ischemic preconditioning and postconditioning. In this review, we discuss the potential application of ischemic preconditioning and postconditioning in the clinical arena of myocardial ischemia and reperfusion, and examine the common signaling pathways by which this might be achieved. PMID- 16258570 TI - Therapy insight: weight-loss surgery and major cardiovascular risk factors. AB - Weight-loss surgery is an effective treatment for severe, medically complicated and refractory obesity. It reverses, eliminates or significantly ameliorates major cardiovascular risk factors related to obesity. In a large proportion of patients, the therapy produces significant weight loss, reduces the risk of disability and premature death, and improves quality of life. Surgical treatment by gastric-restrictive and malabsorptive procedures started several decades ago in the US. Since the 1970s, accrued clinical experience and advances in technology, particularly in minimally invasive surgical approaches, have changed this therapy. Some procedures have evolved, whereas others have become obsolete. Today's weight-loss operations are safe, effective and potentially life-saving options for severely obese cardiology patients. This review describes weight-loss surgery procedures and their effects on cardiovascular risk factors. PMID- 16258569 TI - Drug Insight: statins and stroke. AB - Stroke is the third leading cause of death in the US and a common cause of long term disability worldwide. Ischemic strokes, which are often atherothrombotic, account for more than 80% of all strokes. Current stroke prevention focuses on optimizing the treatment of modifiable risk factors, such as hypertension, diabetes and dyslipidemia. The epidemiologic association between serum cholesterol levels and adjusted stroke rates is not as strong as the link between serum cholesterol levels and coronary heart disease. Clinical trials of 3-hydroxy 3-methylglutaryl-coenzyme A reductase inhibitors (statins), which are potent inhibitors of cholesterol synthesis, have demonstrated, however, a marked reduction in stroke risk in hypercholesterolemic and atherosclerotic individuals, with benefits extending to normocholesterolemic individuals. These findings suggest that statins might have additional effects in stroke protection beyond cholesterol reduction. Because statins inhibit the synthesis of isoprenoid intermediates in the cholesterol biosynthetic pathway, which are important lipid attachments for intracellular signaling molecules, they might have direct noncholesterol-dependent effects on inflammatory and endothelial cells. Here we discuss data from clinical trials assessing the effects of statins on stroke risk, as well as outline the mechanisms underlying the cholesterol-independent effects of statins and provide evidence-based recommendations for stroke prevention, based on achieved serum cholesterol levels in patients at risk of stroke. PMID- 16258571 TI - Technology Insight: transcatheter closure of ventricular septal defects. AB - Transcatheter closure of atrial septal defects has been employed increasingly in the past decade. This technique is now regarded as the treatment of choice for patients with appropriate atrial septal defects. Transcatheter closure of ventricular septal defects (VSDs) has undergone fewer clinical trials, even though VSDs are more common than atrial septal defects. The implanted device does not seem to embolize and complications are few. Decreases in left ventricular and diastolic pressure and improvement of ventricular function have been reported early following device closure, and the left-to-right shunt has been either eliminated or dramatically reduced. In small infants who are in heart failure at a young age and who weigh less than 8 kg, which is below the recommended threshold for device closure, technological advancements in device size and catheter manipulation are needed before VSDs can be closed. A large number of VSDs, particularly if small to medium in size, will become smaller or close spontaneously, making intervention unnecessary. Muscular VSDs have been closed with transcatheter devices for the past 15 years. Although perimembranous defects are more common than muscular defects, they have not become more amenable to closure since the introduction of the Amplatzer VSD occluder device (AGA Medical Corporation, Golden Valley, MN). Previous devices, such as the Rashkind and button devices, have been unsuccessful in attempts at closure of the VSDs because of the proximity of the defects to the aortic valve and potential aortic valve damage. Before the transition is made to routine therapy, large, multicenter trials are justified to test the feasibility, safety and efficacy of nonsurgical closure of VSDs. In this review, I discuss the current applications of transcatheter closure of membranous, perimembranous and muscular VSDs, in particular with Amplatzer devices, and the implications for future development. PMID- 16258572 TI - Technology insight: MRI of the myocardium. AB - MRI is emerging as the method of choice for the evaluation of a wide variety of cardiovascular disorders. A major advantage of this technique over the other cardiac imaging modalities is the fact that it allows the operator--via special software programs called pulse sequences--to probe a vast array of biological properties while using the same machine. In this review, we provide the reader with a brief overview of the pulse sequence concept and how it enables MRI practitioners to pursue a multifaceted approach to evaluating the myocardium. We discuss how MRI technology makes this imaging method ideally suited to the assessment of cardiac morphology, contractile function, myocardial perfusion and infarction. In addition, we present clinical scenarios in which the performance of multifaceted imaging by MRI can alter clinical decision making. PMID- 16258573 TI - Healthline: do primary care doctors agree with the advice? AB - AIMS: To assess agreement between the advice to symptomatic callers to Healthline, and that advised by primary care doctors given the same clinical information, and thus to assess the safety of Healthline advice. METHODS: Ninety records of symptomatic calls to Healthline were examined by three primary care specialists, blinded to the actual advice given. They independently recorded what they would have advised, and their advice was compared with that actually given by the Healthline nurse guided by Care Enhance Call Centre software. RESULTS: Variation among the three doctors was greater than that between the median doctor and Healthline. In 82% of cases, the median doctor triaged to an endpoint close to (or lower than) Healthline. In all but one of the remainder, at least one doctor thought there was no risk to the patient (i.e. in 99% of cases). Review of that case indicated nurse error and the guideline itself was judged to be safe. CONCLUSION: New Zealand primary care specialists regarded the Care Enhance Call Centre decision support software used by Healthline as clinically safe. PMID- 16258574 TI - Teaching imaging to undergraduates: strategies and expectations. PMID- 16258575 TI - Radiology knowledge in new medical graduates in New Zealand. AB - AIMS: To establish the level of knowledge of new medical graduates in New Zealand about common radiological investigations and to assess their ability to request most appropriate, cost-effective radiological investigations for common clinical scenarios. METHODS: A test was developed and administered in Waikato, Christchurch, Rotorua, Auckland, and Dunedin hospitals during the first month of new house officer year (November 2002). RESULTS: Sixty-two first year house officers participated; 59 were New Zealand medical graduates (Auckland: 24 and Otago: 35) and 3 were from overseas institutions. The mean score for questions that assessed about risks involved in common investigations, including radiation, was 47% (95% CI: 45%-49%). The mean score for selecting the appropriate clinical investigations was 53% (95% CI 52%-54%). Most significantly, only 42% (95% CI 38% 46%) of the respondents thought they had adequate radiology teaching to work as house officers. The following percentage of the respondents never observed the respective examination during their medical school training: barium enema 72% (95% CI: 60%-82%); IVU 75% (95% CI: 63%-87%); US scan 25% (95% CI: 16%-37%); CT scan 20% (95% CI: 11%-32%); angiogram 16% (95% CI: 9%-28%); MRI 42% (95% CI: 30% 54%). The mean score for the practical knowledge about common investigations was 50 (95% CI: 48%-52%). CONCLUSIONS: Medical students report that they have limited exposure to radiology teaching during their medical school training. The test results suggest that medical school training enabled them to commence their probationary year with a 'just safe' level of radiology knowledge and skill. PMID- 16258576 TI - The Strong Parents-Strong Children Programme: parental support in serious and chronic child illness. AB - AIM: To determine the effectiveness of an intervention programme developed to help parents manage serious child illness. METHOD: Information from previous research about parents' stress of managing their children's serious illnesses, plus their wishes for what to do about it, were used to develop a behavioural intervention to be used with groups of parents with seriously ill children. The 6 week programme, called 'Strong Parents-Strong Children', was tested using a wait list control design, and evaluated by standardised and researcher-developed psychological measures. RESULTS: Several significant post-test changes in healthier directions were found for the study group, compared with the control group. Additionally, the group process and session helpfulness received positive appraisals, personal goals were attained at high levels, and most participants said they would recommend the programme to others in similar situations. CONCLUSIONS: The programme appears to have a significant and positive impact on the parents of seriously ill children. PMID- 16258577 TI - The health status of quota refugees screened by New Zealand's Auckland Public Health Service between 1995 and 2000. AB - Each year New Zealand accepts approximately 750 refugees from overseas for resettlement in New Zealand. Known as "Quota Refugees", these people arrive in groups of 125 six times each year. Since 1979 their first six weeks in New Zealand have been spent at the Mangere Refugee Resettlement Centre in Auckland. This Centre comprises several agencies which prepare the refugees for their life in New Zealand. Among the agencies is a Medical Clinic, which provides health screening, and management of any medical problems found. This paper describes the findings of the health screening, mainly those refugees screened between 1995 and 2000, but also includes some historical data from the opening of the Resettlement Centre. PMID- 16258578 TI - Computerised screening for hazardous drinking in primary care. AB - INTRODUCTION: Brief interventions undertaken in primary care settings have been shown consistently to reduce hazardous drinking, but they are not commonly offered in practice. The aims were to determine the uptake by young people of an offer of screening in a primary care setting; to identify patients' drinking risk levels; and to estimate the proportion who would consent to computerised brief intervention and follow-up. METHODS: Participants were 1120 patients attending a university student health service that were invited for screening while in the waiting room. Participants were also asked for their consent to be contacted for follow-up assessment 1, 6, and 12 months later. RESULTS: 1,010 patients (90%) accepted the invitation for screening. Of these, 35 (4%) failed to complete screening, thus leaving 975 with complete Alcohol Use Disorders Identification Test (AUDIT) data. Sixty percent of women and 73% of men screened positive. Twenty-three patients (4%) eligible for intervention declined follow-up assessments. DISCUSSION: The study demonstrates that the primary care setting can be used to facilitate access via computer to a large number of individuals whose drinking is hazardous. Limitations of the study include the use of an educated segment of the population who may be more receptive to computerised screening than other groups. Strengths of the study include the high rate of participation and the naturalistic setting in which the data were collected. PMID- 16258579 TI - The efficacy of EMG-biofeedback training on quadriceps muscle strength in patients after arthroscopic meniscectomy. AB - AIMS: In 40 patients, we attempted to investigate the efficacy of electromyography-biofeedback (EMG-B) on quadriceps muscle strength after arthroscopic meniscectomy. METHODS: The patients were randomly divided into two groups each consisting of 20 subjects. For the control group, a classical exercise program was given (five sessions of EMG-B application for 2 weeks postoperatively). Range of motions, Lysholm knee score, EMG electrical activity values of vastus medialis obliques (VMO), and vastus lateralis (VL) were measured pre- and postoperatively on the 3rd and 14th day, and at the 6th week. RESULTS: When the ranges of motion values were compared, a significant difference (for average values of knee flexion angle) was found on the 14th day and 6th week in favour of biofeedback group (p<0.05). When Lysholm knee scores on the 14th day and 6th week were compared in the control and biofeedback groups, and maximum contraction and average contraction values of VMO, VL muscles were compared with operated/non-operated %age ratios, there was a statistically significant difference in favour of the biofeedback group (p<0.05). CONCLUSIONS: Our results showed that EMG-B was an effective treatment modality in improving quadriceps muscle strength after arthroscopic meniscectomy surgery. PMID- 16258580 TI - Metastatic thyroid carcinoma. AB - Although metastases to the thyroid gland are common in autopsy studies, clinically significant metastases are rare. A 58-year-old Turkish patient, presenting with thyroid metastasis 2 years after undergoing left nephrectomy for renal cell carcinoma, is reported in this case report. Thyroid metastasis can be the initial presentation of renal cell carcinoma, or it may occur a long time after nephrectomy, which can lead to misdiagnosis of primary thyroid neoplasm. Radiographic features are not useful in making discrimination between the two, however a fine needle aspiration biopsy can be useful. The role of surgical therapy is controversial. PMID- 16258581 TI - TNF inhibitors for inflammatory arthritis in New Zealand. AB - For the vast majority of the estimated 100,000 New Zealanders who suffer from rheumatoid arthritis (RA), relatively inexpensive disease-modifying antirheumatic drugs (DMARD) regimens are sufficient to control inflammatory disease and maintain long-term function. Some DMARDs have been shown to slow, but not arrest, the progression of erosions. All but a few of those who suffer from ankylosing spondylitis (AS) can manage full social participation with non-steroidal anti inflammatory drugs (NSAIDs) and an exercise regimen. For the small subset of arthritis sufferers who have disabling pain and progressive damage from uncontrolled inflammatory disease, the advent of the biological era offered great promise. In most of the developed world, this promise is being delivered to patients with an expanding range of diseases including RA, AS, and psoriatic arthritis, but central government (PHARMAC) funding for TNF inhibitors in New Zealand has until recently been limited to etanercept for approximately 40 patients with juvenile inflammatory arthritis. PMID- 16258582 TI - Regarding 'access to palliative care for people with motor neurone disease in New Zealand'. PMID- 16258583 TI - Access to palliative care for people with motor neurone disease in New Zealand: a patient's perspective. PMID- 16258584 TI - Re-evaluating a local public health control measure used in New Zealand for the pandemic influenza of 1918. PMID- 16258585 TI - PHARMAC responds on long-acting insulin analogues. PMID- 16258586 TI - More on 'violence against women in New Zealand'. PMID- 16258587 TI - A patient's diary. PMID- 16258588 TI - Blame it on Big Tobacco, but do what you can to help smokers stop. PMID- 16258589 TI - A corpulent heart. Lipomatous hypertrophy of the interatrial septum. PMID- 16258590 TI - Protective effects of nimesulide (COX Inhibitor), AKBA (5-LOX Inhibitor), and their combination in aging-associated abnormalities in mice. AB - Several inflammatory processes play a critical role in brain aging and are associated with increased vulnerability to neurodegeneration. Cyclooxygenase-2 (COX-2) and 5-lipoxygenase (5-LOX), two enzymes involved in the oxygenation of the arachidonic acid, are upregulated in the central nervous system during aging and are associated with different aging-related brain pathologies. The present experiment was performed to study the effects of 5-LOX inhibitor, acetyl-11-keto beta-boswellic acid (AKBA), nimesulide (preferential COX-2 inhibitor), and their combination on cognitive performance of young and aged mice, using elevated plus maze test. Chronic administration of AKBA (100 mg/kg, p.o.) and nimesulide (2.42 mg/kg, p.o.) for 15 days significantly reversed the aging-induced retention deficit in mice. Coadministration of AKBA and nimesulide enhanced the cognitive performance in aged mice when compared with that in per se treatment. The aging related increase in oxidative damage (increased LPO and decreased GSH) was reversed by AKBA, nimesulide, and their combination. Further, per se COX and LOX inhibitors and their combination did not produce any alteration in gastrointestinal parameters; they also reversed the aging-induced motor dysfunction in the aged animals. On the basis of these observations, present findings indicated that the combination of COX and LOX inhibitors (dual inhibitors) may provide a new therapeutic innovation for the treatment of aging related brain disorders such as Alzheimer's disease and different motor dysfunctions with adequate gastrointestinal tolerability. PMID- 16258591 TI - Antidepressant-like effect of hyperbaric oxygen treatment in forced-swimming test in rats. AB - The present study was undertaken to assess the antidepressant-like activity of hyperbaric oxygen (HBO) treatment and also to investigate whether in the forced swimming test HBO treatment interacts with the antidepressant effects of fluoxetine, a selective serotonin reuptake inhibitor, and imipramine, which is mainly a noradrenaline reuptake inhibitor. HBO treatment (at 2.4 atmospheres absolute [ATA] for 60 min) significantly reduced the immobility time in this test; in other words, it displayed antidepressant-like activity. The coadministration of HBO with fluoxetine (5 mg/kg) or imipramine (5 mg/kg) at their ineffective doses produced greater inhibition of immobility time compared with HBO alone. In conclusion, HBO treatment might be an alternative approach to antidepressant therapy, alone or in combination with antidepressant drugs. PMID- 16258592 TI - Effect of progesterone on calcium activated potassium currents and intracellular calcium in guinea pig colon myocytes. AB - AIMS: To study the effects of progesterone on contractile activity of smooth muscle strips and on ion currents and intracellular Ca2+ ([Ca2+]i) intensity in single colonic myocytes in guinea pig proximal colons. METHODS: Strips and single cells were dissected from female guinea pig proximal colon. Contraction of strips through an isotonic transducer was assessed and the responsible currents to progesterone were recorded with EPC-9 amplifier in nystatin perforated whole-cell configuration. Detection of [Ca2+]i fluorescence loading fura-2 acetoxymethylester (fura-2/AM) was measured with confocal microscope. RESULTS: Progesterone significantly inhibited contraction of guinea pig colon strips in a dose-dependent pattern. Inhibitory concentration 50 (IC50) of progesterone in longitudinal strips and circular strips was, respectively, 9.7 microM and 1.0 nM. Iberiotoxin (IbTX) partially blocked inhibition of progesterone in both oriented smooth muscle strips. Ca2+ activated K+ (K(Ca)) channel currents recorded by depolarizing pulse protocol were enhanced by progesterone to 138% +/- 13% (n = 9, p < 0.01), and to 143% +/- 12% (n = 8, p < 0.01) when perfused with 10 mcM onapristone. Progesterone reduced L-Ca2+ currents to 67% +/- 6% (n = 7, p < 0.01) and had no effect with 5 microM nicardipine in bath solution. [Ca2+]i fluorescence was reduced by progesterone to 75% +/- 12% (n = 8, p < 0.01). CONCLUSION: Progesterone decreases the contraction of colonic smooth muscles by enhancing K(Ca) currents and reducing Ca2+ influx. PMID- 16258593 TI - Neuropeptide dietary supplement N-PEP-12 enhances cognitive function and activates brain bioelectrical activity in healthy elderly subjects. AB - N-PEP-12 is a dietary supplement consisting of neuropeptides and amino acids. In animal experiments, the compound has been shown to enhance cognitive function and reduce neurodegenerative events associated with aging. In this study, we investigated the effects of a single oral dose of N-PEP-12 (180 mg) on brain bioelectrical activity and cognitive performance in healthy elderly subjects. N PEP-12 induced a significant (p < 0.05) increase in relative alpha-activity power 6 h after administration. This enhancement was accompanied by a generalized decrease in slow Delta-activity. Significant improvement in memory performance subtests was also seen 6 h after N-PEP-12 administration in some but not in all tests. Taken together, these data suggest that N-PEP-12 might be a reliable dietary supplement to be investigated for improving and, perhaps, maintaining brain function among healthy older adults. PMID- 16258595 TI - Specification of laboratory animal use in scientific articles: current low detail in the journals' instructions for authors and some proposals. AB - The scientific article communicates results of research to other investigators; therefore, it must contain a complete description of the experiment to help other researchers when designing their future investigations. However, poorly detailed data on laboratory animal use is given in published articles. Despite the well known and important contribution of the International Committee of Medical Journal Editors (ICMJE) to standardize scientific writing and submission of manuscripts to biomedical journals, no specific instructions on the reporting of animal use are given in the ICMJE Uniform Requirements and, therefore, most journals do not detail this to the authors. Individual efforts from groups like the Working Committee for the Biological Characterizations of Laboratory Animals, the Boyd Group, or the Committee on Publication Ethics are commendable. These contributions should be incorporated into the ICMJE Uniform Requirements and, later, into the Instructions for Authors of peer-reviewed journals. This would be the only efficient way to instruct authors on how to report laboratory animal use in their submitted manuscripts. The present article relates some proposals for helping authors when reporting animal use in scientific articles. These proposals are not only based on previous guidelines for animal specifications, but also on Instructions for Authors from journals specialized in Laboratory Animal Science. These proposals are classified into major and minor issues, and they are located in the corresponding parts of the article, as defined by the Introduction, Methods, Results, and Discussion (IMRAD) method. PMID- 16258594 TI - Effects of granisetron with droperidol or dexamethasone on prevention of postoperative nausea and vomiting after general anesthesia for cesarean section. AB - This prospective, placebo-controlled, double-blinded, and randomized study was undertaken to compare the efficacy of granisetron, droperidol, and combinations of granisetron with droperidol or dexamethasone on postoperative nausea and vomiting in patients undergoing general anesthesia for cesarean section. Patients (n = 150) who were scheduled for cesarean section under general anesthesia were randomly assigned to one of the five groups: physiological saline 5 ml in Group A, granisetron 40 microg/kg + dexamethasone 8 mg in Group B, granisetron 40 microg/kg + droperidol 1.25 mg in Group C, droperidol 1.25 mg in Group D, and granisetron 40 microg/kg in Group E were administered intravenously after clamping of the fetal umbilical cord. Postoperative nausea and vomiting was observed for 024 h after the anesthesia. Cesarean sections were all performed under general anesthesia. Postoperative nausea and vomiting was more common in placebo group (56.7%) than the others during the 0-24 h after the anesthesia (p < 0.05). All granisetron groups were more effective than placebo and droperidol groups during the postoperative 3-24 h (p < 0.01). Although this trial lacks statistical power, granisetron alone and combinations with droperidol or dexamethasone were effective similarly. All treatment groups, except droperidol during the postoperative 3-24 h, were effective for prevention of postoperative nausea and vomiting during the postoperative 0-24 h. PMID- 16258596 TI - Gateways to clinical trials. AB - Gateways to Clinical Trials are a guide to the most recent clinical trials in the current literature and congresses. The data in the following tables have been retrieved from the Clinical Trials Knowledge Area of Prous Science Integrity, the drug discovery and development portal, http://integrity.prous.com. This issue focuses on the following selection of drugs: AAV-NGFbeta, aprepitant, aripiprazole, atomoxetine hydrochloride; beta-Methyl-6-chloromelatonin, BMS 214662, bortezomib, bosentan; Calcipotriol/betamethasone dipropionate, CEA TRICOM, cetuximab, ciclesonide, clofarabine, Cypher; Dalbavancin, darbepoetin alfa, darifenacin hydrobromide, desloratadine, Dexamet, drospirenone, drospirenone/ethinylestradiol, drotrecogin alfa (activated), duloxetine hydrochloride, dutasteride; Ecogramostim, efalizumab, ertapenem sodium, escitalopram oxalate, eszopiclone; Fenretinide; Gefitinib, gestodene, ghrelin (human); hMaxi-K, human papillomavirus vaccine; Imatinib mesylate, indiplon, iodine (i131) tositumomab, irofulven, ISS-1018; Lasofoxifene tartrate, levodopa/carbidopa/entacapone, liposomal doxorubicin; Nemifitide ditriflutate, nesiritide; Omalizumab; Pegfilgrastim, peginterferon alfa-2a, peginterferon alfa 2b, phVEGF-A165, pimecrolimus, pramlintide acetate; Rasburicase, rimonabant hydrochloride; Satraplatin, St. John's Wort extract, sunitinib malate; Tadalafil, tanaproget, Taxus, tiotropium bromide, treprostinil sodium; Valdecoxib, vardenafil hydrochloride hydrate; Ximelagatran; Zileuton. PMID- 16258597 TI - [Gout's genes and geniouses]. PMID- 16258598 TI - [The bisphosponates: chemical characteristics, skeletal biological effects and extra-skeletal effects]. AB - Bisphosphonates (BP) are pharmacological compounds whose the most important biological effect is the reduction of bone remodelling, explaining the reason for their use in pathological conditions characterised by an increased bone resorption, such as osteoporosis, Paget's disease, malign hypercalcemia during myeloma and osteolytic bone metastasis. Nevertheless there are several experimental evidence that BP possess different extra-skeletal biological effects, ranging from analgesic properties, anti-inflammatory and pro inflammatory effects and the capacity of modifying the biological activity of cells other than osteoblasts and osteoclasts, such as the immune system cells and other cells of mesenchymal origin. Several data report the beneficial effects of BP as anti-inflammatory agents in different inflammatory chronic articular diseases, which make BP suitable drugs for treatment of pathologies other than bone disease. PMID- 16258599 TI - [Comorbidities in rheumatoid arthritis: analysis of hospital discharge records]. AB - OBJECTIVE: Arthritis is often associated with comorbidities. For many of them, such as hypertension, cardiovascular disease, chronic pulmonary disease, and upper gastrointestinal disease, arthritis and its treatment may also represent a risk factor. This study is concerned with an evaluation of the frequency of comorbidities in a cohort of patients with rheumatoid arthritis (RA). METHODS: The discharge diagnoses of patients with RA during the period 1 January 1997 to 31 December 2000 were retrieved from the database of the Department of Internal Medicine of the University of Genova, Italy. The diagnosis of RA was made if the patient's discharge record contained the code 714 of the International Classification of Diseases, IX revision, as first 3 numbers. The other diagnoses were also recorded along with demographic data, type and duration of hospital stay, and performed procedures. RESULTS: During the study period, 427 patients with RA were admitted to the hospital for a total number of 761 admissions, which represented 2.2% of total admissions. Ninety-one (21.3%) patients did not have comorbidities, whereas 336 (78.6%) had one or more comorbidities. The most frequently observed comorbidities were cardiovascular diseases (34.6%), including hypertension (14.5%) and angina (3.5%), followed by gastrointestinal (24.5%), genito-urinary (18.7%) and respiratory (17%) diseases. There was a male predominance (p=0.004) within patients with comorbidities, who were significantly older (64.2+/-3.2 years vs. 57.2+/-4.2 years; p<0.001) and required longer periods of hospital stay (22.7 days vs. 12.5 days; p<0.001). CONCLUSIONS: Comorbidities are present in nearly 80% of RA inpatients. Comorbidity is a good predictor of health outcome, health services utilization, and medical costs. Because RA comorbidity can act as confounder, it should be considered in epidemiologic studies and clinical trials. PMID- 16258600 TI - [Validation of the Italian versions of the Bath Ankylosing Spondylitis Functional Index (BASFI) and the Dougados Functional Index (DFI) in patients with ankylosing spondylitis]. AB - OBJECTIVES: The Bath Ankylosing Spondylitis Functional Index (BASFI) and the Dougados Functional Index (DFI) are the most commonly used instruments to measure functioning in ankylosing spondylitis (AS). The aim of this study was to translate, adapt and validate these instruments into the Italian language. METHODS: The BASFI and DFI questionnaires were translated into Italian by two independent bilingual physicians who were familiar with the medical aspects of AS and by one professional translator. Two rheumatologists familiar with instrument validation, and who were aware of the purpose of the study, examined semantic, idiomatic and conceptual issues and produced by consensus unified versions of each instrument. English back-translations from the Italian were done by a professional translator unaware of the original version. Both English versions were compared, and where needed, modifications to the Italian versions were made. RESULTS: A total of 95 patients were included: 77 males, age (mean+/-SD) 47.9+/ 9.3years, and disease duration 12.4+/-6.6 years, and 18 females, age 45.9+/-8.7 years, and disease duration 11.3+/-8.2 years. Reliability, measured in 23 patients participating a physiotherapy program, showed an acceptable one-week test-retest intraclass correlation coefficient (ICC)--BASFI ICC: 0.91, 95% CI: 0,87-0.94 and DFI ICC: 0.86, 95% CI: 0.83-0.90. The internal consistency was 0.90 (Cronbach's alpha) for the BASFI and 0.87 for the DFI. For validity the functional indices were correlated with the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI), Bath Ankylosing Spondylitis Metrology Index (BASMI), Bath Ankylosing Spondylitis Patient Global Score (BAS-G), modified Health Assesment Questionnaire (HAQ-S), SF-36 physical component summary (SF-36 PCS), stiffness, pain, physician's assessment of disease activity, Bath AS Radiology Index-total (BASRI-t), erythrocyte sedimentation rate (ESR), and C-reactive protein (CRP). The functional indices (BASFI and DFI) were correlated with each other (p<0.0001) and with activity variables. There was no significant relationship between functional indices and BASRI-t and acute phase reactants. The receiver operating characteristic (ROC) curve analysis indicated that the BASFI ranked superior compared to HAQ-S, (p = 0.019) and SF36 PCS (p = 0.002), but not respect to DFI (p = NS), in distinguishing between patients with high and low disease activity. CONCLUSIONS: The Italian versions of the BASFI and DFI showed adequate reliability and validity in patients with AS. Because of psychometric advantages, the BASFI may be preferred in clinical trial settings. However, sensitivity to changes due to drug therapy and/or rehabilitation remains to be determined. PMID- 16258601 TI - [Endothelial progenitor cells in systemic sclerosis: their possible role in angiogenesis]. AB - BACKGROUND: Recently, several studies have demonstrated the presence of circulating endothelial progenitors (CEPs) responsible for angiogenesis. Notably, these cells are able to migrate to ischemic tissues and differentiate in situ in mature endothelial cells. Aim of this study was to assess the presence of CEPs in the peripheral blood of patients with Sistemic Sclerosis (SSc) and evaluate their significance as an attempt of re-vascularization MATERIAL AND METHODS: Samples of peripheral blood from 40 healthy subjects and 56 patients with SSc were studied. Five-parameter, 3-color flow cytometry was performed with a FACScan. CEPs were defined as CD45 negative, CD34 and CD133 positive. In addition, plasma levels of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) were detected by commercial ELISA (R&D Systems). RESULTS: Levels of CEPs (CD133+/CD34+/CD45-) were significantly higher in patients with SSc in comparison to HC (P = 0.01). No correlation was found between CEPs and any clinical parameter of disease neither activity score. CEPs were significantly higher in the group of patients with early disease, while their number decreased in the late phases of disease. Plasma levels of VEGF, but not bFGF, were significantly higher in SSc in comparison to HC (P<0.001) but no correlation was found between VEGF concentrations and CEP number. CONCLUSIONS: The presence of CEPs in patients with SSc suggest that sclerodermic hypoxic tissues could induce the mobilization of bone-marrow derived cells in an attempt to provided new vessels, in the early phase of the disease, at least. PMID- 16258602 TI - [Isolated congenital heart block in undifferentiated connective tissue disease and in primary Sjogren's syndrome: a clinical study of 81 pregnancies in 41 patients]. AB - OBJECTIVE: To study the incidence and the features of congenital heart block (CHB) in patients with undifferentiated connective tissue disease (UCTD) and primary Sjogren's syndrome (pSS). METHODS: We studied 81 pregnancies of 41 women attending the Outpatients' Clinic of the Rheumatology Unit of University Hospital of Padova from July 1989 to March 2004. Twenty five of these (61%) were affected with UCTD and 16 (39%) with pSS. Serologic inclusion criteria was anti-Ro/La positivity, assessed by counterimmunoelectrophoresis and ELISA. RESULTS: CHB was found in 2 out of the 46 (4.3%) pregnancies followed by our Staff and in 2 out of the 35 (5.7%) included in the retrospective part of the study. In 3 cases CHB was a 3rd degree block, causing pregnancy termination in 2. The only 2nd degree block was identified in one patient at the 22nd week of gestation and treated with dexamethasone and plasma-exchange. All of the women were positive to 52 kd and 60 kd Ro autoantibodies. CHB mothers had higher titer antibodies to 52 kd Ro protein than did the mothers with healthy infants (P = 0.026). Electrocardiographic abnormalities at birth were found in 3 out of 29 asymptomatic infants. One presented sinus bradycardia, the second abnormalities of ventricular repolarization, both regressed spontaneously, while the third ventricular extrasystoles which continue even now at 5 months. CONCLUSIONS: These results showed that in UCTD and pSS there is a higher incidence of CHB than that reported in Systemic Lupus Erythematosus. Electrocardiographic screening in all infants born to mothers with anti-Ro/La antibodies would seem an important measure to identify those with irreversible heart conduction abnormalities. PMID- 16258603 TI - [Wegener's granulomatosis: an analysis of 50 patients]. AB - OBJECTIVES: To evaluate the rate of different organs involvement in 50 patients with Wegener's Granulomatosis (GW), and to describe their clinical manifestations and their response to treatment. METHODS: We evaluated 50 consecutive patients with GW, come to our attention from January 1987 to May 2003. 43 patients met the 1990 American College of Rheumatology (ACR) criteria for classification of GW; 7 patients the 1993 ELK criteria. RESULTS: 82% of patients presented Ear/Nose/Throat (ENT) involvement, which is the most common site of inflammation. 22% of our patients had ENT-restricted disease; in 78% of cases disease extended to other organs. Lungs were involved in 72% of cases; kidney in 36%; eye in 24%; nervous system (NS) in 14% (central NS in 10% and peripheral NS in 4%); skin in 10%; heart in 8%; testis in 4%. Arthritis was present in 10% of patients. We discuss treatment of all patients and response to therapy of those 28 whose follow-up is available. CONCLUSIONS: Involvement of airways and kidney is by far the most common in GW, though potentially any other organ or system may be affected. The total rate of other organs involvement is 70%. PMID- 16258604 TI - [17beta-Estradiol and testosterone influence the mRNA expression and the time course of inflammatory cytokines in activated human monocytic cell line (THP-1)]. AB - OBJECTIVE: The aim of this study was to evaluate the effects of 17beta-estradiol (E2) and testosterone (T) on the mRNA expression of IL-1beta, IL-6, TNF-alpha and TGF-beta in cultured human monocytic cells (THP-1) after INF-gamma activation. METHODS: THP-1 were cultured with E2 and T (10 nM) for 24 hs and then activated with INF-gamma (500 U/ml), during different periods of time (1, 3, 6, and 12 hs). After total RNA extraction, all samples were analyzed by multiple RT-PCR to detect mRNA expression of the selected cytokines. RESULTS: Cells cultured without hormonal treatment expressed IL-1beta mRNA after 1 h; on the contrary TNF-alpha, TGF-beta and IL-6 mRNA were expressed only after 3 hs. At 6 and 12 hs only IL-6 mRNA was still expressed. Interestingly, cells cultured with testosterone never expressed IL-1beta nor TNF-alpha mRNA and showed an IL-6 mRNA expression similar to the untreated controls at 3, 6 and 12 hours. On the contrary, cells treated with E2 showed the expression of all cytokines at 3 and 12 hs, and in general showed an higher expression of all the analyzed cytokines mRNA when compared to the other conditions. CONCLUSIONS: This study suggests that sex hormones may modulate the cytokine mRNA expression in the inflammatory cells. In fact, T inhibits TNF-alpha production at all the tested times, whereas E2 seems to accelerate and to enhance the inflammatory response. Therefore, the altered sex hormone ratio, as observed in the synovial fluid of RA patients (high E2/low T), might contribute to the occurrence and last of synovitis. PMID- 16258605 TI - [Tenosynovitis of the ankles as onset of sarcoidosis in a patient with ulcerative colitis]. AB - Arthritis and tenosynovitis are frequently reported as complications of inflammatory bowel diseases. About 10% of patients with ulcerative colitis presents articular inflammation, usually in the phases of activity of intestinal disease. Tenosynovitis is also a frequent complication of ulcerative colitis. We describe here a case of tenosynovitis of both ankles occurring in a patient affected by ulcerative colitis not in active phase. Chest X-ray and TC showed hilar lymph node enlargement and transbronchial biopsy confirmed the diagnosis of sarcoidosis. In this disease tenosynovitis is very rare, unlike arthritis that is rather common. In conclusion we observed a case of ankle bilateral tenosynovitis as onset manifestation of sarcoidosis. PMID- 16258606 TI - [Temporomandibular joint involvement in juvenile idiopathic arthritis: treatment with an orthodontic appliance]. AB - OBJECTIVE: About 65% of children suffering from juvenile idiopathic arthritis (JIA) shows a more or less marked involvement of temporo-mandibular joint (TMJ) with altered mandibular growth, resorption of the condyles, occlusary instability, reduced chewing ability and facial dysmorphia. The purpose of our study is to prevent and to treat the progressive evolution of JIA on craniofacial growth and morphology with a functional appliance; surgery should be considered only in so far as the adequacy of TMJ movement is concerned. METHODS: From 1992 until now 72 children with proved JIA and TMJ involvement have been treated (50 females, 22 males, aged 6 to 16 years old). TMJ involvement was bilateral in 61% and unilateral in 39% of patients. A diagnostic workup was carried out involving tomograms of TMJ and cephalometric radiograph and analysis. The authors used a bimaxillary activator in the attempt to modify the unfavourable growth pattern and provide a gradual ante-rotation of the jaw. RESULTS: Almost all JIA patients showed satisfactory long term results, easing of pain, reduced skeletal discrepancy, increased function and good facial profile. CONCLUSIONS: The long term results of this study indicate that orthopaedic therapy might control the vicious circle of the malocclusion in children with JIA, preventing exacerbation of mandibular clockwise rotation. Surgical intervention for the improvement of TMJ function should be considered only if a severe restricted state is imminent. PMID- 16258607 TI - [Detection and identification of crystals in synovial fluid]. AB - Synovial fluid analysis for crystals represents one of the most important laboratory test for the evaluation of rheumatic diseases. The identification of monosodium urate and calcium pyrophosphate dihydrate crystals allows the prompt diagnosis of gout and pyrophosphate crystal-related arthropaties. Crystals are identified based on their shape and birefringence through a polarized light microscope equipped with a first order red compensator. Due to its simple execution and high diagnostic value, this examine should be always performed to complete synovial fluid analysis. PMID- 16258608 TI - Formation of protein-coated iron minerals. AB - The ability of iron to cycle between Fe(2+) and Fe(3+) forms has led to the evolution, in different forms, of several iron-containing protein cofactors that are essential for a wide variety of cellular processes, to the extent that virtually all cells require iron for survival and prosperity. The redox properties of iron, however, also mean that this metal is potentially highly toxic and this, coupled with the extreme insolubility of Fe(3+), presents the cell with the significant problem of how to maintain this essential metal in a safe and bioavailable form. This has been overcome through the evolution of proteins capable of reversibly storing iron in the form of a Fe(3+) mineral. For several decades the ferritins have been synonymous with the function of iron storage. Within this family are subfamilies of mammalian, plant and bacterial ferritins which are all composed of 24 subunits assembled to form an essentially spherical protein with a central cavity in which the mineral is laid down. In the past few years it has become clear that other proteins, belonging to the family of DNA-binding proteins from starved cells (the Dps family), which are oligomers of 12 subunits, and to the frataxin family, which may contain up to 48 subunits, are also able to lay down a Fe(3+) mineral core. Here we present an overview of the formation of protein-coated iron minerals, with particular emphasis on the structures of the protein coats and the mechanisms by which they promote core formation. We show on the one hand that significant mechanistic similarities exist between structurally dissimilar proteins, while on the other that relatively small structural differences between otherwise similar proteins result in quite dramatic mechanistic differences. PMID- 16258609 TI - Group 4 salicyloxazolines are potent polymerization catalysts. AB - Octahedral titanium and zirconium complexes based on salicyloxazoline ligands with sterically demanding ortho-substituents provide a new family of extremely active ethene polymerization catalysts [up to 10(8) g PE (mol bar h)(-1)] which are in some cases "single site". PMID- 16258610 TI - Novel rhodium and ruthenium carbonyl cluster complexes with face- and edge bridging GaCp* ligands. Synthesis and structural characterization of the Rh6(CO)12(mu3-GaCp*)4 and Ru6(eta6-C)(mu2-CO)(CO)13(mu3-GaCp*)2(mu2-GaCp*) clusters. AB - Reactions of hexanuclear carbonyl clusters of rhodium Rh(6)(CO)(16) and ruthenium Ru(6)(eta(6)-C)(micro(2)-CO)(CO)(16) with GaCp*(Cp*= C(5)Me(5)) in the mild conditions result in substitution of CO ligands and formation of the Rh(6)(CO)(12)(micro(3)-GaCp*)(4) and the Ru(6)(eta(6)-C)(micro(2) CO)(CO)(13)(micro(3)-GaCp*)(2)(micro(2)-GaCp*) cluster derivatives. PMID- 16258611 TI - Synthesis and characterization of seven-coordinate tripodal imidazole complexes of iron(II) and manganese(II). AB - The iron(II) and manganese(II) complexes of the N(7) Schiff-base condensate of tris(3-aminopropyl)amine with 1-methyl-2-imidazolecarbaldehyde and the manganese(II) complex of the N(7) Schiff-base condensate of tris(3 aminopropyl)amine with 4-imidazolecarbaldehyde are high-spin mono capped octahedral seven-coordinate complexes with a short, approximately 2.44 e, metal to apical nitrogen bond. PMID- 16258612 TI - Calix[4]phyrin based redox architectures: towards new molecular tools for electrochemical sensing. AB - New redox active molecular macrocyclic architectures characterized by a direct connection between dipyrrin, tripyrrin and ferrocenyl fragments have been synthesized and characterized. Contrarily to fully conjugated porphyrins, in which four pyrrole moieties contribute to the overall aromatic pi-electronic system and behave as a unique electroactive species, calixphyrins can be regarded as an assembly of independent redox active pyrrole and conjugated oligopyrrole fragments linked through sp(3) hybridized meso carbon atoms. The disruption of the conjugation pathway not only multiplies the number of redox centres throughout the molecule but also leads to a large variety of molecular architectures with specific physico-chemical properties. These novel ferrocene containing hybrid macrocycles exhibit especially attractive electronic and structural features suited for use as molecular sensing tools. An efficient voltammetric sensing of exogenic electron rich anionic species could especially be performed using a metallo-calix[4]phyrin-(1.1.1.1) through the displacement of the labile axial binding site, the perturbation of the Fc(0/+) redox couple being directly related to complexed species features. PMID- 16258613 TI - Pi-conjugated N-heterocyclic compounds: correlation of computational and electrochemical data. AB - Electrochemical reduction potentials of a broad selection of nitrogen-containing molecules suitable as bridging (dipodal and tripodal) ligands in coordination and organometallic chemistry are reported and compared with results of semiempirical calculations. Trends of electrode potentials observed experimentally agree with respective calculated data, deviations can be explained by invoking peculiarities of the involved molecular orbitals and ligand-electrode surface interactions. PMID- 16258614 TI - Molecular modelling for coordination compounds: Cu(II)-amine complexes. AB - The Ligand Field Molecular Mechanics (LFMM) method has been applied to 85 Cu(II) amine complexes, eighteen of which were selected to form a training set. A single set of parameters yields Cu-N bond lengths for four-, five- and six-coordinate systems generally within 0.04 A of their X-ray crystallographic values. Larger deviations are indicative of counterion effects and/or crystallographic artefacts arising from Jahn-Teller averaging. The LFMM treatment is flexible and unbiased and for simple ligands, automatically gives planar CuN(4) and tetragonally elongated CuN(6) complexes. In agreement with experiment, square-pyramidal coordination is marginally favoured over trigonal bipyramidal coordination for CuN(5) species. However, if the ligand requirements dictate, the LFMM accommodates trigonal bipyramidal coordination for CuN(5) species, tetrahedral distortions of CuN(4) species and cis-elongated CuN(6) species. PMID- 16258615 TI - Single-strand helical complexes constructed from 2-pyridinyl-3 pyridinylmethanone: tuning the helical pitch length by variation of metal cation and/or counter anion. AB - The new ligand 2-pyridinyl-3-pyridinylmethanone (L) proves to be an excellent building block for the construction of single-strand helical architectures. A series of helical complexes have been synthesized by the reaction of L with various metal salts, in which L exhibits three kinds of coordination modes involving two kinds of bridging conformations, resulting in four types of single strand helical chains. The counter anions in the series of 2(1) helical silver(I) complexes {[Ag(L)]X}(infinity)(X = NO(3), 1; PF(6), 2; BF(4), 3; ClO(4), 4; CF(3)CO(2), 5; CF(3)SO(3), 6) are fully or partially embedded inside the cylindrical helix, and the pitch length corresponds not only to the size of the anion but also to its manner of docking into the groove of the helix. Formation of the helical structure in {[Cu(L)(CH(3)CN)(H(2)O)(ClO(4))]ClO(4)}(infinity)(7) is driven by Ow-H...O (perchlorate) hydrogen bonding that leads to a stable triangular motif which rigidly fixes the configuration of the helix. In {[Co(L)(H(2)O)(3)](ClO(4))(2).2H(2)O}(infinity)(8) and {[Zn(L)(H(2)O)(3)](CF(3)SO(3))(2).H(2)O}(infinity)(9), similar helical chains without anion embedment suggest that the pitch length can be tuned by the size of metal cations. Notably, complex {[Ag(L)]CF(3)SO(3)}(infinity)(10), a conformational polymorph of , has a 4(1) helix induced by argentophilic interaction. PMID- 16258616 TI - Acid-base, coordination and oxidative properties of systems containing ATP, L histidine and Ni(II) ions. AB - Potentiometric measurements of ATP-His system proved an existence of five adducts in the solution with stoichiometries ranging from H(5)(ATP)(His) to H(ATP)(His)(4 ). Their formation is a consequence of electrostatic interactions only. In the ternary Ni(II)-ATP-His system, two complex species NiH(ATP)(His)(2-) and Ni(ATP)(His)(3-), were found. In the former, stacking interaction between the aromatic moiety of ATP and the imidazole ring of l-histidine is crucial to the adduct stability. All studied systems are able to generate single strand lesions of plasmid DNA in the presence of hydrogen peroxide. However, only binary systems produce linear form of DNA, which is a consequence of the accumulation of the single-stranded breaks. PMID- 16258617 TI - Speciation in the aqueous H+/H2VO4-/H2O2/phosphate system. AB - The speciation in the aqueous H(+)/H(2)VO(4)(-)/phosphate (dihydrogen phosphate, P(-)) and H(+)/H(2)VO(4)(-)/H(2)O(2)/P(-) systems has been determined in the physiological medium of 0.150 M Na(Cl) at 25 degrees C. A combination of multinuclear NMR integral and chemical shift (Bruker AMX500) as well as potentiometric data (glass electrode) have been collected and treated simultaneously by the computer program LAKE. The pK(a)-values for phosphoric acid have been determined by potentiometric and (31)P NMR chemical shift data, and have been found to be 1.85 +/- 0.02, 6.69 +/- 0.02 and 11.58 +/- 0.07. The errors given are 3sigma. Altogether nine vanadate-phosphate species have been found in the ternary H(+)/H(2)VO(4)(-)/P(-) system in the pH region 1-11, with the following compositions: VP, VP(2) and V(14)P. Equilibrium is very slow in acidic solutions, requiring more than 3 months for the formation of V(14)P species. On the other hand, less than 15 min are needed for equilibration at neutral and alkaline pH. In the quaternary H(+)/H(2)VO(4)(-)/H(2)O(2)/P(-) system, four new species have been found in addition to all binary and ternary complexes. They are of VXP and VX(2)P compositions, where X denotes the peroxo ligand. (51)V and (31)P NMR chemical shifts, compositions and formation constants are given, and equilibrium conditions are illustrated in distribution diagrams as well as the fit of the model to the experimental data. Biological and medical relevance of the species is also discussed and physiological conditions are modelled. PMID- 16258618 TI - Proton exchange kinetics from the bound waters on the oxo-centered rhodium(III) trimer [Rh3(mu3-O)(mu-O2CCH3)6(OH2)3]+: a variable pH and temperature 1H NMR study. AB - Proton exchange from the bound to the bulk waters on the oxo-centered rhodium(III) trimer, [Rh(3)(micro(3)-O)(micro O(2)CCH(3))(6)(OH(2))(3)](+)(abbreviated as Rh(3)(+)), was investigated over the temperature range of 219.1-313.9 K using a (1)H NMR line-broadening technique. By solving the modified Bloch equations for a two-site chemical exchange, lifetimes (tau) for proton transfer at pH = 2.7, 3.6, and 7.0 ([Rh(3)(+)]= 26 mM, T= 298 K) were determined to be 0.3 (+/-.08) ms, 2 (+/-0.3) ms, and 0.2 (+/-0.2) ms, respectively. From the temperature dependence of the rate, the activation parameters were determined to be DeltaH(++)= 16.2 (+/-0.5) kJ mol(-1) and DeltaS(++)=- 123 (+/-2) J mol(-1) K(-1), DeltaH(++)= 14.9 (+/-0.5) kJ mol(-1) and DeltaS(++)=- 141 (+/-2) J mol(-1) K(-1), and DeltaH(++)= 45 (+/-2) kJ mol(-1) and DeltaS(++)=- 22 (+/-5) J mol(-1) K(-1) for pH = 2.7, 3.6 and 7.0, respectively. All results are reported for a mixed solvent system [acetone : 250 mM NaClO(4)(aq)(3:1)], which was necessary to depress the freezing point of the solution so that the (1)H NMR signal due to bound water could be observed. The pK(a) of Rh(3)(+) was measured to be 8.9 (+/-0.2) in the mixed solvent, which is near the pK(a) for an aqueous solution (8.3 (+/-0.2)). Surprisingly, the lifetimes for protons on Rh(3)(+) are close to those observed for the Rh(OH(2))(6)(3+) ion, in spite of the considerable difference in structure, Bronsted acidity of the bound waters and average charge on the metal ion. PMID- 16258619 TI - Alkyne coupling at a rhenium-monocarborane substrate: synthesis of Re,B eta2:sigma-butadienyl complexes. AB - Treatment of 7-NH(2)Bu(t)-nido-7-CB(10)H(12) in tetrahydrofuran (THF) with LiBu(n)(3 equiv) and then [ReBr(CO)(3)(THF)(2)] gives the rhenacarborane dianion [1-NHBu(t)-2,2,2-(CO)(3)-closo-2,1-ReCB(10)H(10)](2-), isolated as the bis [N(PPh(3))(2)](+) salt (4). Iodine oxidation of this Re(I) intermediate gives the Re(III) complex [1,2-mu-NHBu(t)-2,2,2-(CO)(3)-closo-2,1-ReCB(10)H(10)] 6 in which the carborane functions formally as an 8-electron (6pi+ 2sigma) donor. Reaction of with ligands L in the presence of Me(3)NO gives substituted products [1,2-mu NHBu(t)-2,2-(CO)(2)-2-L-closo-2,1-ReCB(10)H(10)][L = PPh(3)(7a), CNXyl (7b; Xyl = C(6)H(3)Me(2)-2,6), or Bu(t)C triple bond CH (7c)]. Formation of complex 7c is unexpectedly accompanied by [1,2-mu-NHBu(t)-2,2-(CO)(2)-3,2-sigma:eta(2) {C(=CHBu(t))-CH=CHBu(t)}-closo-2,1-ReCB(10)H(9)] 8a, in which an alkyne-derived dienyl group is bound to both the rhenium centre and to an adjacent boron vertex. Complex 8a is also obtained from 7c with Bu(t)C triple bond CH and Me(3)NO. The same reaction of 7c, using PhC triple bond CH or CNXyl instead of Bu(t)C triple bond CH, gives, respectively, [1,2-micro-NHBu(t)-2,2-(CO)(2)-3,2-sigma:eta(2) {C(=CHBu(t))-CH=CHPh}-closo-2,1-ReCB(10)H(9)] 8b and [1,2-micro-NHBu(t)-2-Bu(t)C triple bond CH-2-CO-2-CNXyl-closo-2,1-ReCB(10)H(10)] 9. Addition of donors L to results in displacement from rhenium of the pendant dienyl moiety, yielding [1,2 mu-NHBu(t)-2,2-(CO)(2)-2-L-3-{C(=CHBu(t))-CH=CHBu(t)}-closo-2,1-ReCB(10)H(9)][L = PMe(3)(10a), CNBu(t)(10b)]. Single-crystal X-ray diffraction analyses have confirmed the novel structural features of compounds 6, 7c, 8b and 9. PMID- 16258620 TI - Supramolecular isomerism within three-dimensional 3-connected nets: unusual synthesis and characterization of trimorphic copper(I) 3,5-dimethyl-1,2,4 triazolate. AB - Solvothermal treatments of acetonitrile, aqueous ammonia and copper(II) salts frequently yielded alpha-[Cu(mtz)](1)(Hmtz = 3,5-dimethyl-1,2,4-triazole) under different reaction conditions. However, two new supramolecular isomers of 1, namely beta-[Cu(mtz)](2) and gamma-[Cu(mtz)](3) were successfully isolated upon introducing 4,4'-bipyridine or nitrate as an additive. 1-3 represent the first example of true supramolecular isomerism within 3D 3-connected nets. Besides the 4.8.10 net for the tetragonal phase 1 (P4(2)/n, a= 13.470(2), c= 6.142(2)A), two novel 3-connected nets, 8(2)10-a and 6.10(2), were observed in the new tetragonal phase 2 (I42d, a= 14.0247(5), c= 5.2884(4)A) and orthorhombic phase 3 (Iba2, a= 8.0423(7), b= 11.5310(10), c= 17.3505(16)A), respectively. The three isomers also displayed distinct physical properties related to their different supramolecular structures. PMID- 16258621 TI - C-F Bond activation at Ni(0) and simple reactions of square planar Ni(II) fluoride complexes. AB - The reaction of Ni(COD)(2)(COD = 1,5-cyclooctadiene) with triethylphosphine and pentafluoropyridine in hexane has been shown previously to yield trans-[NiF(2 C(5)NF(4))(PEt(3))(2)](1a) with a preference for reaction at the 2-position of the heteroaromatic. The corresponding reaction with 2,3,5,6-tetrafluoropyridine was shown to yield trans-[NiF(2-C(5)NF(3)H)(PEt(3))(2)](1b). In this paper, we show that reaction of Ni(COD)(2) with triethylphosphine and pentafluoropyridine in THF yields a mixture of 1a and 1b. Competition reactions of Ni(COD)(2) with triethylphosphine in the presence of mixtures of heteroaromatics in hexane reveal a kinetic preference of k(pentafluoropyridine):k(2,3,5,6-tetrafluoropyridine)= 5.4:1. Treatment of 1a and 1b with Me(3)SiN(3) affords trans-[Ni(N(3))(2 C(5)NF(4))(PEt(3))(2)](2a) and trans-[Ni(N(3))(2-C(5)NHF(3))(PEt(3))(2)](2b), respectively. The complex trans-[Ni(NCO)(2-C(5)NHF(3))(PEt(3))(2)](3b) is obtained on reaction of with Me(3)SiNCO and by photolysis of under CO, while trans-[Ni(eta(1)-C [triple bond CPh)(2-C(5)NF(4))(PEt(3))(2)](4a) is obtained by reaction of phenylacetylene with 1a. Addition of KCN, KI and NaOAc to complex 1a affords trans-[Ni(X)(2-C(5)NF(4))(PEt(3))(2)](5a X = CN, 6a X = I, 7a X = OAc), respectively. The PEt(3) groups of complex are readily replaced by addition of 1,2-bis(dicyclohexylphosphino)ethane (dcpe) to produce [NiF(2 C(5)F(4)N)(dcpe)](8a). Addition of dcpe to trans-[Ni(OTf)(2 C(5)F(4)N)(PEt(3))(2)](10a), however, yields the salt [Ni(2 C(5)F(4)N)(dcpe)(PEt(3))](OTf)(9a) by substitution of only one PEt(3) and displacement of the triflate ligand. The structures of 2b, 4a, 7a and 8a were determined by X-ray crystallography. The influence of different ancillary ligands on the bond lengths and angles of square-planar nickel structures with polyfluoropyridyl ligands is analysed. PMID- 16258622 TI - Synthesis, structures, magnetism and electrochemical properties of triruthenium acetylide complexes. AB - A series of triruthenium complexes with arylacetylide axial ligands Ru(3)(dpa)(4)(C(2)X)(2)(BF(4))(y)(dpa = dipyridylamido; X = Fc, y= 0 (1); X = Ph, y= 0 (2); X = PhOCH(3), y= 1 (3); X = PhC(5)H(11), y= 1 (4); X = PhCN, y= 0 (5); X = PhNO(2), y= 0 (6)) have been synthesized. The crystal structures show that the Ru-Ru bond lengths (2.3304(9)-2.3572(5)A) of these compounds are longer than those of Ru(3)(dpa)(4)Cl(2)(Ru-Ru=2.2537(1)A). This is ascribed to the formation of the stronger pi-backbonding from metal to axial ligand which weakens the Ru-Ru interactions and the bond order is reduced in the triruthenium unit. Cyclic voltammetry and differential pulse voltammetry show that compound exhibits electronic coupling between the two ferrocenyl units with DeltaE(1/2) close to 100 mV. Compounds 2-6 display three triruthenium-based reversible one-electron redox couples, two oxidations and one reduction, and the electrode potentials shift upon varying the substituents. A linear relationship is observed when the Hammett constants are plotted against the redox potentials. PMID- 16258623 TI - An acid-sensing ion channel that detects ischemic pain. AB - Ischemic pain occurs when there is insufficient blood flow for the metabolic needs of an organ. The pain of a heart attack is the prototypical example. Multiple compounds released from ischemic muscle likely contribute to this pain by acting on sensory neurons that innervate muscle. One such compound is lactic acid. Here, we show that ASIC3 (acid-sensing ion channel #3) has the appropriate expression pattern and physical properties to be the detector of this lactic acid. In rats, it is expressed only in sensory neurons and then only on a minority (approximately 40%) of these. Nevertheless, it is expressed at extremely high levels on virtually all dorsal root ganglion sensory neurons that innervate the heart. It is extraordinarily sensitive to protons (Hill slope 4, half activating pH 6.7), allowing it to readily respond to the small changes in extracellular pH (from 7.4 to 7.0) that occur during muscle ischemia. Moreover, both extracellular lactate and extracellular ATP increase the sensitivity of ASIC3 to protons. This final property makes ASIC3 a "coincidence detector" of three molecules that appear during ischemia, thereby allowing it to better detect acidosis caused by ischemia than other forms of systemic acidosis such as hypercapnia. PMID- 16258624 TI - The GATO gene annotation tool for research laboratories. AB - Large-scale genome projects have generated a rapidly increasing number of DNA sequences. Therefore, development of computational methods to rapidly analyze these sequences is essential for progress in genomic research. Here we present an automatic annotation system for preliminary analysis of DNA sequences. The gene annotation tool (GATO) is a Bioinformatics pipeline designed to facilitate routine functional annotation and easy access to annotated genes. It was designed in view of the frequent need of genomic researchers to access data pertaining to a common set of genes. In the GATO system, annotation is generated by querying some of the Web-accessible resources and the information is stored in a local database, which keeps a record of all previous annotation results. GATO may be accessed from everywhere through the internet or may be run locally if a large number of sequences are going to be annotated. It is implemented in PHP and Perl and may be run on any suitable Web server. Usually, installation and application of annotation systems require experience and are time consuming, but GATO is simple and practical, allowing anyone with basic skills in informatics to access it without any special training. GATO can be downloaded at [http://mariwork.iq.usp.br/gato/]. Minimum computer free space required is 2 MB. PMID- 16258625 TI - Indole ring oxidation by activated leukocytes prevents the production of hypochlorous acid. AB - Hypochlorous acid (HOCl) released by activated leukocytes has been implicated in the tissue damage that characterizes chronic inflammatory diseases. In this investigation, 14 indole derivatives, including metabolites such as melatonin, tryptophan and indole-3-acetic acid, were screened for their ability to inhibit the generation of this endogenous oxidant by stimulated leukocytes. The release of HOCl was measured by the production of taurine-chloramine when the leukocytes (2 x 10(6) cells/mL) were incubated at 37 degrees C in 10 mM phosphate-buffered saline, pH 7.4, for 30 min with 5 mM taurine and stimulated with 100 nM phorbol 12-myristate acetate. Irrespective of the group substituted in the indole ring, all the compounds tested including indole, 2-methylindole, 3-methylindole, 2,3 dimethylindole, 2,5-dimethylindole, 2-phenylindole, 5-methoxyindole, 6 methoxyindole, 5-methoxy-2-methylindole, melatonin, tryptophan, indole-3-acetic acid, 5-methoxy-2-methyl-3-indole-acetic acid, and indomethacin (10 microM) inhibited the chlorinating activity of myeloperoxidase (MPO) in the 23-72% range. The compounds 3-methylindole and indole-3-acetic acid were chosen as representative of indole derivatives in a dose-response study using purified MPO. The IC50 obtained were 0.10 +/- 0.03 and 5.0 +/- 1.0 microM (N = 13), respectively. These compounds did not affect the peroxidation activity of MPO or the production of superoxide anion by stimulated leukocytes. By following the spectral change of MPO during the enzyme turnover, the inhibition of HOCl production can be explained on the basis of the accumulation of the redox form compound-II (MPO-II), which is an inactive chlorinating species. These results show that indole derivatives are effective and selective inhibitors of MPO chlorinating activity. PMID- 16258626 TI - Secretion of Streptomyces tendae antifungal protein 1 by Lactococcus lactis. AB - Lactococcus lactis, the model lactic acid bacterium, is a good candidate for heterologous protein production in both foodstuffs and the digestive tract. We attempted to produce Streptomyces tendae antifungal protein 1 (Afp1) in L. lactis with the objective of constructing a strain able to limit fungal growth. Since Afp1 activity requires disulfide bond (DSB) formation and since intracellular redox conditions are reportedly unfavorable for DSB formation in prokaryotes, Afp1 was produced as a secreted form. An inducible expression-secretion system was used to drive Afp1 secretion by L. lactis; Afp1 was fused or not with LEISSTCDA, a synthetic propeptide (LEISS) that has been described to be a secretion enhancer. Production of Afp1 alone was not achieved, but production of LEISS-Afp1 was confirmed by Western blot and immunodetection with anti-Afp1 antibodies. This protein (molecular mass: 9.8 kDa) is the smallest non bacteriocin heterologous protein ever reported to be secreted in L. lactis via the Sec-dependent pathway. However, no anti-fungal activity was detected, even in concentrated samples of induced supernatant. This could be due to a too low secretion yield of Afp1 in L. lactis, to the absence of DSB formation, or to an improper DSB formation involving the additional cysteine residue included in LEISS propeptide. This raises questions about size limits, conformation problems, and protein secretion yields in L. lactis. PMID- 16258627 TI - Thermodynamic evaluation and modeling of proton and water exchange associated with benzamidine and berenil binding to beta-trypsin. AB - Serine-proteases are involved in vital processes in virtually all species. They are important targets for researchers studying the relationships between protein structure and activity, for the rational design of new pharmaceuticals. Trypsin was used as a model to assess a possible differential contribution of hydration water to the binding of two synthetic inhibitors. Thermodynamic parameters for the association of bovine beta-trypsin (homogeneous material, observed 23,294.4 +/- 0.2 Da, theoretical 23,292.5 Da) with the inhibitors benzamidine and berenil at pH 8.0, 25 degrees C and with 25 mM CaCl2, were determined using isothermal titration calorimetry and the osmotic stress method. The association constant for berenil was about 12 times higher compared to the one for benzamidine (binding constants are K = 596,599 +/- 25,057 and 49,513 +/- 2,732 M(-1), respectively; the number of binding sites is the same for both ligands, N = 0.99 +/- 0.05). Apparently the driving force responsible for this large difference of affinity is not due to hydrophobic interactions because the variation in heat capacity (DeltaCp), a characteristic signature of these interactions, was similar in both systems tested (-464.7 +/- 23.9 and -477.1 +/- 86.8 J K(-1) mol(-1) for berenil and benzamidine, respectively). The results also indicated that the enzyme has a net gain of about 21 water molecules regardless of the inhibitor tested. It was shown that the difference in affinity could be due to a larger number of interactions between berenil and the enzyme based on computational modeling. The data support the view that pharmaceuticals derived from benzamidine that enable hydrogen bond formation outside the catalytic binding pocket of beta-trypsin may result in more effective inhibitors. PMID- 16258628 TI - Symmetric pollen mitosis I and suppression of pollen mitosis II prevent pollen development in Brachiaria jubata (Gramineae). AB - Microsporogenesis and pollen development were analyzed in a tetraploid (2n = 4x = 36) accession of the forage grass Brachiaria jubata (BRA 007820) from the Embrapa Beef Cattle Brachiaria collection that showed partial male sterility. Microsporocytes and pollen grains were prepared by squashing and staining with 0.5% propionic carmine. The meiotic process was typical of polyploids, with precocious chromosome migration to the poles and laggards in both meiosis I and II, resulting in tetrads with micronuclei in some microspores. After callose dissolution, microspores were released into the anther locule and appeared to be normal. Although each microspore initiated its differentiation into a pollen grain, in 11.1% of them nucleus polarization was not observed, i.e., pollen mitosis I was symmetric and the typical hemispherical cell plate was not detected. After a central cytokinesis, two equal-sized cells showing equal chromatin condensation and the same nuclear shape and size were formed. Generative cells and vegetative cells could not be distinguished. These cells did not undergo the second pollen mitosis and after completion of pollen wall synthesis each gave rise to a sterile and uninucleate pollen grain. The frequency of abnormal pollen mitosis varied among flowers and also among inflorescences. All plants were equally affected. The absence of fertile sperm cells in a considerable amount of pollen grains in this accession of B. jubata may compromise its use in breeding and could explain, at least in part, why seed production is low when compared with the amount of flowers per raceme. PMID- 16258629 TI - Plasmablastic multiple myeloma is associated with increased vascular endothelial growth factor immunoexpression. AB - The biologic basis of the negative prognosis of plasmablastic myeloma is not fully understood. To determine whether histologically aggressive multiple myeloma (MM) is associated with a more angiogenic marrow environment, bone marrow samples from 50 recently diagnosed MM patients were evaluated. Twelve percent (6/50) of patients presented plasmablastic MM, and this feature correlated with moderate/strong intensity of vascular endothelial growth factor staining of plasma cells (P = 0.036). Although plasmablastic MM was not associated with increasing of microvessel density, this new evidence of increased expression of vascular endothelial growth factor on plasmablasts suggests that the adverse prognosis conferred by plasmablastic disease may be due, at least in part, to secretion of this angiogenic cytokine, also suggesting that the subset of MM patients with plasmablastic features may derive particular benefit from antiangiogenic therapies. PMID- 16258630 TI - Distribution of microglial cells in the cerebral hemispheres of embryonic and neonatal chicks. AB - The distribution, morphology and morphometry of microglial cells in the chick cerebral hemispheres from embryonic day 4 (E4) to the first neonatal day (P1) were studied by histochemical labeling with a tomato (Lycopersicon esculentum) lectin. The histochemical analysis revealed lectin-reactive cells in the nervous parenchyma on day E4. Between E4 (5.7 +/- 1.35 mm length) and E17 (8.25 +/- 1.2 mm length), the lectin-reactive cells were identified as ameboid microglia and observed starting from the subventricular layer, distributed throughout the mantle layer and in the proximity of the blood vessels. After day E13, the lectin reactive cells exhibited elongated forms with small branched processes, and were considered primitive ramified microglia. Later, between E18 (5.85 +/- 1.5 mm cell body length) and P1 (3.25 +/- 0.6 mm cell body length), cells with more elongated branched processes were observed, constituting the ramified microglia. Our findings provide additional information on the migration and differentiation of microglial cells, whose ramified form is observed at the end of embryonic development. The present paper focused on the arrangement of microglial cells in developing cerebral hemispheres of embryonic and neonatal chicks, which are little studied in the literature. Details of morphology, morphometry and spatial distribution of microglial cells contributed to the understanding of bird and mammal central nervous system ontogeny. Furthermore, the identification and localization of microglial cells during the normal development could be used as a morphological guide for embryonic brain injury researches. PMID- 16258631 TI - Use of blends of bioabsorbable poly(L-lactic acid)/poly(hydroxybutyrate- co hydroxyvalerate) as surfaces for Vero cell culture. AB - Vero cells, a cell line established from the kidney of the African green monkey (Cercopithecus aethiops), were cultured in F-10 Ham medium supplemented with 10% fetal calf serum at 37 degrees C on membranes of poly(L-lactic acid) (PLLA), poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) and their blends in different proportions (100/0, 60/40, 50/50, 40/60, and 0/100). The present study evaluated morphology of cells grown on different polymeric substrates after 24 h of culture by scanning electron microscopy. Cell adhesion was also analyzed after 2 h of inoculation. For cell growth evaluation, the cells were maintained in culture for 48, 120, 240, and 360 h. For cytochemical study, the cells were cultured for 120 or 240 h, fixed, processed for histological analysis, and stained with Toluidine blue, pH 4.0, and Xylidine ponceau, pH 2.5. Our results showed that cell adhesion was better when 60/40 and 50/50 blends were used although cells were able to grow and proliferate on all blends tested. When using PLLA/PHBV (50/50) slightly flattened cells were observed on porous and smooth areas. PLLA/PHBV (40/60) blends presented flattened cells on smooth areas. PLLA/PHBV (0/100), which presented no pores, also supported spreading cells interconnected by thin filaments. Histological sections showed that cells grew as a confluent monolayer on different substrates. Cytochemical analysis showed basophilic cells, indicating a large amount of RNA and proteins. Hence, we detected changes in cell morphology induced by alterations in blend proportions. This suggests that the cells changed their differentiation pattern when on various PLLA/PHBV blend surfaces. PMID- 16258632 TI - Perspectives of digestive pest control with proteinase inhibitors that mainly affect the trypsin-like activity of Anticarsia gemmatalis Hubner (Lepidoptera: Noctuidae). AB - The present study describes the main characteristics of the proteolytic activities of the velvetbean caterpillar, Anticarsia gemmatalis Hubner, and their sensitivity to proteinase inhibitors and activators. Midguts of last instar larvae reared on an artificial diet were homogenized in 0.15 M NaCl and centrifuged at 14,000 g for 10 min at 4 degrees C and the supernatants were used in enzymatic assays at 30 degrees C, pH 10.0. Basal total proteolytic activity (azocasein hydrolysis) was 1.14 +/- 0.15 absorbance variation min(-1) mg protein( 1), at 420 nm; basal trypsin-like activity (N-benzoyl-L-arginine-p-nitroanilide, BApNA, hydrolysis) was 0.217 +/- 0.02 mmol p-nitroaniline min(-1) mg protein(-1). The maximum proteolytic activities were observed at pH 10.5 using azocasein and at pH 10.0 using BApNA, this pH being identical to the midgut pH of 10.0. The maximum trypsin-like activity occurred at 50 degrees C, a temperature that reduces enzyme stability to 80 and 60% of the original, when pre-incubated for 5 and 30 min, respectively. Phenylmethylsulfonyl fluoride inhibited the proteolytic activities with an IC50 of 0.39 mM for azocasein hydrolysis and of 1.35 mM for BApNA hydrolysis. Benzamidine inhibited the hydrolysis with an IC50 of 0.69 and 0.076 mM for azocasein and BApNA, respectively. The absence of cysteine proteinases is indicated by the fact that 2-mercaptoethanol and L-cysteine did not increase the rate of azocasein hydrolysis. These results demonstrate the presence of serine-proteinases and the predominance of trypsin-like activity in the midgut of Lepidoptera insects, now also detected in A. gemmatalis, and suggest this enzyme as a major target for pest control based on disruption of protein metabolism using proteinase inhibitors. PMID- 16258633 TI - Human T-cell lymphotropic virus type I (HTLV-I) proviral DNA viral load among asymptomatic patients and patients with HTLV-I-associated myelopathy/tropical spastic paraparesis. AB - To evaluate the human T-cell lymphotropic virus type I (HTLV-I) proviral DNA load among asymptomatic HTLV-I-infected carriers and patients with HTLV-I-associated myelopathy/tropical spastic paraparesis (HAM/TSP), real time PCR using TaqMan probes for the pol gene was performed in two million peripheral blood mononuclear cells (PBMC). The albumin gene was the internal genomic control and MT2 cells were used as positive control. The results are reported as copies/10,000 PBMC, and the detection limit was 10 copies. A total of 89 subjects (44 HAM/TSP and 45 healthy HTLV-I-infected carriers) followed up at the Institute of Infectious Diseases "Emilio Ribas" and in the Neurology Division of Hospital of Clinicas were studied. The asymptomatic HTLV-I-infected carriers had a median number of 271 copies (ranging from 5 to 4756 copies), whereas the HAM/TSP cases presented a median of 679 copies (5-5360 copies) in 10,000 PBMC. Thus, HAM/TSP patients presented a significantly higher HTLV-I proviral DNA load than healthy HTLV-I carriers (P = 0.005, one-way Mann-Whitney test). As observed in other persistent infections, proviral DNA load quantification may be an important tool for monotoring HTLV-I-infected subjects. However, long-term follow-up is necessary to validate this assay in the clinical setting. PMID- 16258634 TI - Ginkgo biloba leaf extract (EGb 761) enhances catalepsy induced by haloperidol and L-nitroarginine in mice. AB - Ginkgo biloba extract EGb 761 has been reported to have therapeutic effects which have been attributed to anti-oxidant and free radical-scavenging activities, including a direct action on nitric oxide production. L G-nitro-arginine (L NOARG), a nitric oxide synthase inhibitor, and haloperidol, a drug that blocks dopamine receptors, are both known to induce catalepsy in rodents. Nitric oxide has been shown to influence dopaminergic transmission in the striatum. The purpose of the present study was to evaluate the effect of the extract obtained from leaves of Ginkgo biloba tree EGb 761 on catalepsy induced by haloperidol or by L-NOARG. Albino Swiss mice (35-45 g, N = 8-12) received by gavage a single or repeated oral dose (twice a day for 4 days) of EGb 761 followed by ip injection of haloperidol or L-NOARG. After the treatments, the animals were submitted to behavioral evaluation using the catalepsy test. Acute treatment with 80 mg/kg EGb did not modify the catalepsy induced by L-NOARG but, the dose of 40 mg/kg significantly enhanced haloperidol-induced catalepsy measured at the 10th min of the test. After repeated treatment with 80 mg/kg EGb 761, a significant increase in the cataleptic effect produced by both haloperidol and L-NOARG was observed. These data show that repeated EGb 761 administration increases the effects of drugs that modify motor behavior in mice. Since the catalepsy test has predictive value regarding extrapyramidal effects, the possibility of pharmacological interactions between haloperidol and Ginkgo biloba extracts should be further investigated in clinical studies. PMID- 16258635 TI - The validity and 4-year test-retest reliability of the Brazilian version of the Eating Attitudes Test-26. AB - In a cross-sectional study conducted four years ago to assess the validity of the Brazilian version of the Eating Attitudes Test-26 (EAT-26) for the identification of abnormal eating behaviors in a population of young females in Southern Brazil, 56 women presented abnormal eating behavior as indicated by the EAT-26 and the Edinburgh Bulimic Investigation Test. They were each matched for age and neighborhood to two normal controls (N = 112) and were re-assessed four years later with the two screening questionnaires plus the Composite International Diagnostic Interview (CIDI). The EAT results were then compared to diagnoses originating from the CIDI. To evaluate the temporal stability of the two screening questionnaires, a test-retest design was applied to estimate kappa coefficients for individual items. Given the prevalence of eating disorders of 6.2%, the CIDI psychiatry interview was applied to 161 women. Of these, 0.6% exhibited anorexia nervosa and 5.6%, bulimia nervosa (10 positive cases). The validity coefficients of the EAT were: 40% sensitivity, 84% specificity, and 14% positive predictive value. Cronbach's coefficient was 0.75. For each EAT item, the kappa index was not higher than 0.344 and the correlation coefficient was lower than 0.488. We conclude that the EAT-26 exhibited low validity coefficients for sensitivity and positive predictive value, and showed a poor temporal stability. It is reasonable to assume that these results were not influenced by the low prevalence of eating disorders in the community. Thus, the results cast doubts on the ability of the EAT-26 test to identify cases of abnormal eating behaviors in this population. PMID- 16258636 TI - An empirical comparison of atypical bulimia nervosa and binge eating disorder. AB - The International Classification of Diseases, 10th edition (ICD-10) defines atypical bulimia nervosa (ABN) as an eating disorder that encompasses several different syndromes, including the DSM-IV binge eating disorder (BED). We investigated whether patients with BED can be differentiated clinically from patients with ABN who do not meet criteria for BED. Fifty-three obese patients were examined using the Structured Clinical Interview for DSM-IV and the ICD-10 criteria for eating disorders. All volunteers completed the Binge Eating Scale (BES), the Beck Depression Inventory, and the Symptom Checklist-90 (SCL-90). Individuals fulfilling criteria for both ABN and BED (N = 18), ABN without BED (N = 16), and obese controls (N = 19) were compared and contrasted. Patients with ABN and BED and patients with ABN without BED displayed similar levels of binge eating severity according to the BES (31.05 +/- 7.7 and 30.05 +/- 5.5, respectively), which were significantly higher than those found in the obese controls (18.32 +/- 8.7; P < 0.001 and P < 0.001, respectively). When compared to patients with ABN and BED, patients with ABN without BED showed increased lifetime rates of agoraphobia (P = 0.02) and increased scores in the somatization (1.97 +/- 0.85 vs 1.02 +/- 0.68; P = 0.001), obsessive-compulsive (2.10 +/- 1.03 vs 1.22 +/- 0.88; P = 0.01), anxiety (1.70 +/- 0.82 vs 1.02 +/- 0.72; P = 0.02), anger (1.41 +/- 1.03 vs 0.59 +/- 0.54; P = 0.005) and psychoticism (1.49 +/- 0.93 vs 0.75 +/- 0.55; P = 0.01) dimensions of the SCL-90. The BED construct may represent a subgroup of ABN with less comorbities and associated symptoms. PMID- 16258637 TI - Chronic excitotoxic lesion of the dorsal raphe nucleus induces sodium appetite. AB - We determined if the dorsal raphe nucleus (DRN) exerts tonic control of basal and stimulated sodium and water intake. Male Wistar rats weighing 300-350 g were microinjected with phosphate buffer (PB-DRN, N = 11) or 1 microg/0.2 microl, in a single dose, ibotenic acid (IBO-DRN, N = 9 to 10) through a guide cannula into the DRN and were observed for 21 days in order to measure basal sodium appetite and water intake and in the following situations: furosemide-induced sodium depletion (20 mg/kg, sc, 24 h before the experiment) and a low dose of dietary captopril (1 mg/g chow). From the 6th day after ibotenic acid injection IBO-DRN rats showed an increase in sodium appetite (12.0 +/- 2.3 to 22.3 +/- 4.6 ml 0.3 M NaCl intake) whereas PB-DRN did not exceed 2 ml (P < 0.001). Water intake was comparable in both groups. In addition to a higher dipsogenic response, sodium depleted IBO-DRN animals displayed an increase of 0.3 M NaCl intake compared to PB-DRN (37.4 +/- 3.8 vs 21.6 +/- 3.9 ml 300 min after fluid offer, P < 0.001). Captopril added to chow caused an increase of 0.3 M NaCl intake during the first 2 days (IBO-DRN, 33.8 +/- 4.3 and 32.5 +/- 3.4 ml on day 1 and day 2, respectively, vs 20.2 +/- 2.8 ml on day 0, P < 0.001). These data support the view that DRN, probably via ascending serotonergic system, tonically modulates sodium appetite under basal and sodium depletion conditions and/or after an increase in peripheral or brain angiotensin II. PMID- 16258638 TI - Neo-Timm staining in the thalamus of chronically epileptic rats. AB - The thalamus is an important modulator of seizures and is severely affected in cholinergic models of epilepsy. In the present study, chronically epileptic rats had their brains processed for neo-Timm and acetylcholinesterase two months after the induction of status epilepticus with pilocarpine. Both controls and pilocarpine-treated animals presented neo-Timm staining in the anterodorsal nucleus, laterodorsal nucleus, reticular nucleus, most intralaminar nuclei, nucleus reuniens, and rhomboid nucleus of the thalamus, as well as in the zona incerta. The intensity of neo-Timm staining was similar in control and pilocarpine-treated rats, except for the nucleus reuniens and the rhomboid nucleus, which had a lower intensity of staining in the epileptic group. In animal models of temporal lobe epilepsy, zinc seems to modulate glutamate release and to decrease seizure activity. In this context, a reduction of neo-Timm stained terminals in the midline thalamus could ultimately result in an increased excitatory activity, not only within its related nuclei, but also in anatomical structures that receive their efferent connections. This might contribute to the pathological substrate observed in chronic pilocarpine-treated epileptic animals. PMID- 16258639 TI - Effects of type of physical exercise and leisure activities on the depression scores of obese Brazilian adolescent girls. AB - Several studies have indicated that depressive states may lead to hypokinesia with diminished metabolic rate and energy use. Hypokinesia associated with certain eating behaviors may lead to an unfavorable energy balance that can contribute to the emergence and prevalence of obesity among children and adults. The purpose of the present study was to examine the possibility of reducing depression inventory scores in female adolescents with third-degree obesity while testing the effectiveness of different exercise programs in reducing anxiety and depression scores. The sample consisted of 40 female subjects (mean age 16 +/- 1.56 years) divided into 4 groups (aerobic training, anaerobic training, leisure activities, and control). Subjects had a body mass index of 95% or more in relation to the 50th percentile. The aerobic program consisted of three ergometric bicycle sessions per week over a 3-month period (12 weeks) and the activities were prescribed after determining the anaerobic ventilatory threshold (VO2 threshold). Anaerobic training was based on the Wingate anaerobic power test. The leisure program consisted of a varied range of activities (games, exercises, etc.). A nutritionist interviewed the members of these two groups and the control group every week in order to adapt them to the nutritional guidelines proposed for the study. The study showed that all three programs (aerobic exercise, anaerobic exercise and leisure activities) were effective in reducing body mass. However, we found a significant reduction when analyzing the depression scores only for aerobic exercise (18.9 +/- 9.33 to 10.6 +/- 9.56 or 43.9%) but no significant alterations for anaerobic exercise (11.36 +/- 5.23 to 9.63 +/- 4.78 or 15.22%) and leisure (17.28 +/- 7.55 to 15.07 +/- 7.54 or 12.78%), thus indicating that in principle this type of activity could be included to improve emotional well-being of obese adolescent girls. PMID- 16258640 TI - Routine post-weaning handling of rats prevents isolation rearing-induced deficit in prepulse inhibition. AB - Rats reared under isolation conditions from weaning present a number of behavioral changes compared to animals reared under social conditions (group housing). These changes include deficits in prepulse inhibition (PPI) of the startle reflex to a loud sound. PPI refers to the reduction of the magnitude of the startle reflex when a relatively weak stimulus (the prepulse) precedes by an appropriate time interval the intense startle-elicing stimulus (the pulse). PPI is useful for studying sensorimotor integration. The present study evaluated the effect of handling on the impairment of PPI induced by isolation-rearing. Male Wistar rats (N = 11-15/group) were housed in groups (5 per cage and handled three times a week) or isolated (housed individually) since weaning (21 days) for 10 weeks when they reach approximately 150 g. The isolated rats were divided into "minimally handled" animals (handled once a week for cleaning purposes only) or "handled" animals (handled three times a week). This handling consisted of grasping the rat by the tail and moving it to a clean cage (approximately 5 s). A statistically significant reduction (52%) in the PPI test was found only in the isolated group with minimal handling while no difference was seen between grouped animals and isolated handled animals. These results indicate that isolation rearing causes disruption in the PPI at adult age, which serves as an index of attention deficit. This change in the sensory processing of information induced by post-weaning isolation can be prevented by handling during the development of the animal. PMID- 16258641 TI - The inactivation of the basolateral nucleus of the rat amygdala has an anxiolytic effect in the elevated T-maze and light/dark transition tests. AB - Pharmacological evidence indicates that the basolateral nucleus of the amygdala (BLA) is involved in the mediation of inhibitory avoidance but not of escape behavior in the elevated T-maze test. These defensive responses have been associated with generalized anxiety disorder (GAD) and panic disorder, respectively. In the present study, we determined whether the BLA plays a differential role in the control of inhibitory avoidance and escape responses in the elevated T-maze. Male Wistar rats (250-280 g, N = 9-10 in each treatment group) were pre-exposed to one of the open arms of the maze for 30 min and 24 h later tested in the model after inactivation of the BLA by a local injection of the GABA A receptor agonist muscimol (8 nmol in 0.2 microL). It has been shown that a prior forced exposure to one of the open arms of the maze, by shortening latencies to withdrawal from the open arm during the test, improves the escape task as a behavioral index of panic. The effects of muscimol in the elevated T maze were compared to those caused by this GABA agonist in the avoidance reaction generated in the light/dark transition test. This defensive behavior has also been associated with GAD. In the elevated T-maze, intra-BLA injection of muscimol impaired inhibitory avoidance (control: 187.70 +/- 14.90 s, muscimol: 37.10 +/- 2.63 s), indicating an anxiolytic effect, without interfering with escape performance. The drug also showed an anxiolytic effect in the light/dark transition test as indicated by the increase in the time spent in the lighted compartment (control: 23.50 +/- 2.45 s, muscimol: 47.30 +/- 4.48 s). The present findings point to involvement of the BLA in the modulation of defensive responses that have been associated with GAD. PMID- 16258642 TI - Serum and cerebrospinal fluid concentrations of E-selectin in patients with aneurysmal subarachnoid hemorrhage. AB - The goal of the present study was to determine concentrations of E-selectin in both cerebrospinal fluid (CSF) and serum of patients with aneurysmal subarachnoid hemorrhage (SAH) and to evaluate the correlation between the clinical parameters and E-selectin levels. Both CSF and serum samples obtained from 12 patients with aneurysmal SAH and 8 patients with hydrocephalus (control group) without any other known central nervous system disease were assayed for E-selectin by quantitative enzyme-linked immunosorbent assay and the results were compared between the two groups. Mean levels of soluble forms of E-selectin within the first 3 days and on the 5th and 7th days of SAH were 4.0 +/- 7.9, 2.8 +/- 5.2, and 3.1 +/- 4.9 ng/ml in the patient's CSF, and 33.7 +/- 9.2, 35.1 +/- 7.0, and 35.2 +/- 8.7 ng/ml in serum, respectively. In contrast, mean E-selectin levels were 0.1 +/- 0.2 ng/ml in CSF and 8.7 +/- 5.0 ng/ml in serum of control patients. The difference between groups was statistically significant regarding both CSF and serum E-selectin levels (P < 0.05). Thus, we have demonstrated a marked increase of E-selectin concentration in both CSF and serum of patients with aneurysmal SAH compared with control and suggest that blocking the interaction between E-selectin and vascular endothelium may have a beneficial effect on vasospasms. PMID- 16258643 TI - Quantification of fecal estradiol and progesterone metabolites in Syrian hamsters (Mesocricetus auratus). AB - Alternative methods to the utilization of laboratory animal blood and its by products are particularly attractive, especially regarding hamsters due to their small size and difficulties in obtaining serial blood samples. Steroid hormone metabolite quantification in feces, widely used in studies of free-ranging or intractable animals, is a non-invasive, non-stressor, economical, and animal saving technique which allows longitudinal studies by permitting frequent sampling of the same individual. The present study was undertaken to determine the suitability of this method for laboratory animals. Estradiol and progesterone metabolites were quantified by radioimmunoassay in feces of intact, sexually mature female Syrian hamsters during the estrous cycle (control) and in feces of superovulated females. Metabolites were extracted by fecal dilution in ethanol and quantified by solid phase radioimmunoassay. Median estrogen and progesterone concentrations were 9.703 and 180.74 ng/g feces in the control group, respectively. Peaks of estrogen (22.44 +/- 4.54 ng/g feces) and progesterone (655.95 +/- 129.93 ng/g feces) mean fecal concentrations respectively occurred 12 h before and immediately after ovulation, which is easily detected in this species by observation of a characteristic vaginal postovulatory discharge. Median estrogen and progesterone concentrations (28.159 and 586.57 ng/g feces, respectively) were significantly higher in superovulated animal feces (P < 0.0001). The present study demonstrated that it is possible to monitor ovarian activity in Syrian hamsters non-invasively by measuring fecal estradiol and progesterone metabolites. This technique appears to be a quite encouraging method for the development of new endocrinologic studies on laboratory animals. PMID- 16258644 TI - Functional mapping of the cardiorespiratory effects of dorsal and median raphe nuclei in the rat. AB - The dorsal (DRN) and median (MRN) raphe nuclei are important sources of serotonergic innervation to the forebrain, projecting to sites involved in cardiovascular regulation. These nuclei have been mapped using electrical stimulation, which has the limitation of stimulating fibers of passage. The present study maps these areas with chemical stimulation, investigating their influence on cardiorespiratory parameters. Urethane-anesthetized (1.2 g/kg, iv) male Wistar rats (280-300 g) were instrumented for pulsatile and mean blood pressure (MBP), heart rate, renal nerve activity, and respiratory frequency recordings. Microinjections of L-glutamate (0.18 M, 50-100 nl with 1% Pontamine Sky Blue) were performed within the DRN or the MRN with glass micropipettes. At the end of the experiments the sites of microinjection were identified. The majority of sites within the MRN (86.1%) and DRN (85.4%) evoked pressor responses when stimulated (DRN: DeltaMBP = +14.7 +/- 1.2; MRN: DeltaMBP = +13.6 +/- 1.3 mmHg). The changes in renal nerve activity and respiratory rate caused by L glutamate were +45 +/- 11 and +42 +/- 9% (DRN; P < 0.05%), +40 +/- 10 and +29 +/- 7% (MRN, P < 0.05), respectively. No significant changes were observed in saline microinjected animals. This study shows that: a) the blood pressure increases previously observed by electrical stimulation within the raphe are due to activation of local neurons, b) this pressor effect is due to sympathoexcitation because the stimulation increased renal sympathetic activity but did not produce tachycardia, and c) the stimulation of cell bodies in these nuclei also increases the respiratory rate. PMID- 16258645 TI - Topiramate for the treatment of juvenile myoclonic epilepsy. AB - OBJECTIVE: The aim of this study was to evaluate the efficacy and tolerability of topiramate (TPM) in juvenile myoclonic epilepsy (JME). METHOD: We assessed seizure control and adverse effects of TPM in 22 patients (18 females) aged 13 to 53 years. Target TPM dosage was up to 200 mg/day. The patients were subdivided into 3 groups: those treated with seizure control plus side effects (n=4); treated with non-controlled seizures (n=15) and with JME newly diagnosed (n=3). RESULTS: Sixteen patients completed the first year of the follow-up. Generalized tonic-clonic seizures were completely controlled in 10 (62.5%); more than 50% of reduction in 4 (25.0%) and less than 50% in 2 (12.5%). Myoclonia were controlled in 11 (68.8%) and persisted in 5 (31.2%) patients. Absence seizures were present in 5 (22.7%) of whom 2 (9.0%) showed more than 50% of seizure reduction while 3 (13.6%) presented worsening. Discontinuations were due to inadequate seizure control and adverse events (N=4), low compliance and loss of follow-up (N=2) and subject choice (N=1). CONCLUSION: TPM showed to be an effective and well tolerated drug in the treatment of JME. Although frequently observed, TPM side effects were tolerable and the drug could be maintained in the majority of patients. PMID- 16258646 TI - Tolerability, adverse events and compliance to glatiramer acetate in 28 patients with multiple sclerosis using the drug continuously for at least six months. AB - AIM: To assess tolerability, adverse events and compliance to treatment with glatiramer acetate in multiple sclerosis. METHOD: Review of patient records and individual interviews. RESULTS: 30 individuals residing in the coastal region of the State of Sao Paulo who had been in use of glatiramer acetate for at least 6 months were identified. From this group, 28 individuals came to regular consultations and were individually assessed, their complaints being noted down in confidential records. Ten patients reported systemic reactions to the drug. Four of them stopped the medication due to such reactions. Eight patients reported local reactions to the injections. Compliance with injections was achieved, although three patients reported forgetting the injection on a few days. CONCLUSION: We noticed a higher level of systemic adverse events in our patients than in reports in the literature. PMID- 16258647 TI - Description and characteristics of 81 patients attending the Reference Center for Multiple Sclerosis of the coastal region of the state of Sao Paulo-Brazil. AB - OBJECTIVE: To register multiple sclerosis (MS) patients residing in the coastal region of the State of Sao Paulo (CEREM Litoral Paulista). METHOD: Individual interviews with identified cases of MS. RESULTS: 81 individuals with diagnosis of MS agreed to come for registration (62 females [76.5%], 19 males [23.5%]). 65% of all patients were residents of the city of Santos. The mean age of these patients was 43 years (14 to 74 years), and the Expanded Disability Status Scale (EDSS) was < or = 5.5 in 76.5% of the cases. 82.7% of the assessed patients presented the relapsing/remitting form of MS. 81.5% of all patients were undergoing treatment with immunomodulators. CONCLUSION: Due to their clinical profile, patients seem to come to CEREM Litoral Paulista for prescription of immunomodulators. There is a clear need to identify other cases in the region and to allow other forms of treatment to be put into practice. PMID- 16258648 TI - Capsaicin effects on blinking. AB - Blinking is a normal human phenomenon involving trigeminal and facial pathways. To gain understanding on the neurobiology of blinking, five normal subjects were investigated before and after application of transdermal capsaicin at the forehead for two weeks. No effects of topical capsaicin were detected in eye blink rates. However, when capsaicin was applied to a female subject with blepharospasm, she showed a dramatic restoration of her vision subsequent to blinking modification. Deactivation of abnormal A-to-C fibers cross talks at the trigeminal-facial pathways seems to be the most likely mechanism of such improvement. PMID- 16258649 TI - Dexmedetomidine for awake craniotomy without laryngeal mask. AB - OBJECTIVE: This paper reports the use of dexmedetomidine in three epileptic patients with cavernous angiomas that underwent awake surgery in order to map their speech areas. METHOD: Loading dose of dexmedetomidine varied from 1 microg/Kg/h to 3 microg/Kg/h over 20 minutes and maintenance dose from 0.4 microg/Kg/h to 0.8 microg/Kg/h. RESULTS: There was no occurrence of hemodynamic instability, convulsions or respiratory depression. Patients tolerated well the procedure. CONCLUSION: Dexmedetomidine was useful for awake craniotomy as it decreased patients level of consciousness but did not produce agitation. Laryngeal mask was not necessary to keep air ventilation. PMID- 16258650 TI - Menstrual cycle worsening of epileptic seizures in women with symptomatic focal epilepsy. AB - INTRODUCTION: Hormonal fluctuation is responsible for worsening of epileptic seizures during the menstrual cycle. OBJECTIVE: To identify irregularities in the menstrual cycles of women with mesial temporal lobe epilepsy (MTLE) and extratemporal focal epilepsy (ETFE) and correlate the frequency of seizures during the menstrual cycles. METHOD: We evaluated prospectively women in the menacme with MTLE and ETFE. Calendars were provided for these patients, and they were asked to mark their seizure frequency according to the menses. Calendars were reviewed in each routine medical appointment. RESULTS: Thirty-nine patients with MTLE and 14 with ETFE were evaluated. We registered 211 cycles in the patients with MTLE and 49 in those with ETFE. Irregular menstrual cycles were found in 28 (28/39, 71.7%) patients with MTLE and 6 (6/14, 42.8%) with ETFE (p=0.052). Premenstrual seizure worsening was observed in 46 (21.8%) patients with MTLE and 9 (18.3%) with ETFE (p=0.596). Menstrual worsening was observed in 47 (22.2%) patients with MTLE and 15 (30.6%) with ETFE (p=0.217). Ovulatory worsening was observed in 36 (17%) patients with MTLE and 13 (26.5%) with ETFE (p=0,126). Catamenial worsening was observed in 58 (27.4%) of the patients with MTLE and in 17 (34.7%) of the patients with ETFE (p=0.315). CONCLUSION: There was no difference between the group of patients with MTLE and ETFE regarding the frequency of irregular cycles and seizure worsening during the premenstrual, menstrual, catamenial or ovulatory periods. PMID- 16258651 TI - Seizure occurrence in patients with chronic renal insufficiency in regular hemodialysis program. AB - Hemodialysis-associated seizure is a complication of hemodialysis. This report describes the occurrence of seizures in patients with end stage renal disease on dialysis therapy at the Nephrology Institute of Mogi das Cruzes, Sao Paulo State, Brazil. A retrospective medical history of 189 patients was reviewed to investigate the occurrence of convulsive seizures during dialytic program. Seven patients with history of seizures were selected but five of them were included in our study. Three patients presented generalized tonic-clonic seizures, one had partial seizure with secondary generalization, and one presented unclassified seizure. Three patients presented seizure just during the dialysis (unique seizure) and one of them presented convulsive status epilepticus. The two other patients had already presented seizures prior the beginning of dialysis. We conclude that seizures in renal failure could be considered as occasional events that do not usually become chronic. PMID- 16258652 TI - Sleep disorders are associated with impulsivity in school children aged 8 to 10 years. AB - CONTEXT: Sleep has an important function in the physical and emotional development of children. Some studies suggest an association between impulsivity and sleep disorders. However, little is known about this association in schoolchildren aged 8 to 10 years. METHOD: We studied 1180 children, 547 with sleep disorders (SD) and 633 without SD (control group), classified with SD questionnaires. Within the SD group, 53 children with sleep-related respiratory disorders (SRRD) and 521 children with non-respiratory sleep disorders (NRSD) were analyzed. We assessed emotional indicators of impulsivity with Bender test. RESULTS: More SD children presented impulsivity than control group (p<0.05). More NRSD and 10 years old children presented impulsivity than control group of the same age (p=0.001). Impulsivity and SRRD were associated with 8 years old children (p<0.05). CONCLUSION: Children with SD, 8 years old children with SRRD, and 10 years old children with NRSD presented higher proportion of impulsivity than control children. PMID- 16258653 TI - Depression in Parkinson's disease: study of 60 cases. AB - Depression is very frequent in Parkinson's disease (PD) and largely unrecognized by neurologists, emphasizing the need of an approach to psychiatric symptoms by non psychiatrists in order to ensure an early diagnosis of depression in PD; clinical characteristics and the prevalence rate of depression in PD were evaluated and the relationship of depression in PD with other variables were determined. Sixty PD subjects, who fulfilled the clinical criteria for primary PD, 56.6% males, age range from 44 to 85 years old, in different stages of the disease were investigated. All subjects were submitted to the UPDRS-III, V and VI, Clinical Interview Schedule and the Hamilton depression scale. A significant correlation was found between depression and UPDRS-III, V and VI, anxiety and irritability. The frequency of depression in PD in this study was nearly 40% possessing specific features. Structured interviews and evaluation scales are essential for an accurate diagnosis and proper treatment of depression in PD. PMID- 16258654 TI - Guillain-Barre syndrome in the elderly: clinical, electrophysiological, therapeutic and outcome features. AB - There are few papers devoted to geriatric Guillain-Barre (GBS) and many related issues remain unanswered. OBJECTIVE: To describe clinical, electrophysiological and therapeutic features in this age. METHOD: Clinico-epidemiological data and therapy of GBS patients older than 60 years were reviewed. Hughes scores were used to quantify neurological deficit and define outcome. RESULTS: Among 18 patients (mean age 64.8 years), 9 had evident prodrome and 80% noticed initially sensory-motor deficit. Demyelinating GBS was found in 8 and axonal in 6 subjects. There was one Miller-Fisher and 3 unclassified cases. Plasmapheresis (PFX) was single therapy in 12 patients and intravenous immunoglobulin (IVIg) in 2. Disability scores just before therapy were similar in both groups, so as short and long term outcome. CONCLUSION: Axonal GBS seems to be more frequent in the elderly and this may have prognostic implications. PFX and IVIg were suitable options, but complications were noticed with PFX. Prospective studies are needed to better understand and manage GBS in the elderly. PMID- 16258655 TI - Intercostal nerve mononeuropathy: study of 14 cases. AB - This retrospective study describes 14 cases of intercostal nerve mononeuropathy (INM) found in 5,560 electromyography (EMG) exams performed between January 1991 and June 2004 in our University Hospital. Medical charts of all patients with history of thoracic pain and EMG diagnosis of intercostal mononeuropathy were reviewed. INM was detected in 14 patients; etiology was thoracic surgery in 6 (43%), post-herpetic neuropathy in 4 (28%), probable intercostal neuritis in 2 (14%), lung neoplasia in 1 (7%), and radiculopathy in 1 (7%). From this study, trauma and infection were the main etiologies in intercostal neuropathic pain development. Tricyclic antidepressants and anticonvulsants were the most common therapeutic drugs used. PMID- 16258656 TI - Mild hypothermia reduces polymorphonuclear leukocytes infiltration in induced brain inflammation. AB - Over the last 50 years deep hypothermia (23 degrees C) has demonstrated to be an excellent neuroprotective agent in cerebral ischemic injury. Mild hypothermia (31 33 degrees C) has proven to have the same neuroprotective properties without the detrimental effects of deep hypothermia. Mechanisms of injury that are exaggerated by moderate hyperthermia and ameliorated by hypothermia include, reduction of oxygen radical production, with peroxidase damage to lipids, proteins and DNA, microglial activation and ischemic depolarization, decrease in cerebral metabolic demand for oxygen and reduction of glycerin and excitatory amino acid (EAA) release. Studies have demonstrated that inflammation potentiates cerebral ischemic injury and that hypothermia can reduce neutrophil infiltration in ischemic regions. To further elucidate the mechanisms by which mild hypothermia produces neuroprotection in ischemia by attenuating the inflammatory response, we provoked inflammatory reaction, in brains of rats, dropping a substance that provokes a heavy inflammatory reaction. Two groups of ten animals underwent the same surgical procedure: the skull bone was partially removed, the duramater was opened and an inflammatory substance (5% carrageenin) was topically dropped. The scalp was sutured and, for the group that underwent neuroprotection, an ice bag was placed covering the entire skull surface, in order to maintain the brain temperature between 29.5-31 degrees C during 120 minutes. After three days the animals were sacrificed and their brains were examined. The group protected by hypothermia demonstrated a remarkable reduction of polymorphonuclear leukocytes (PMNL) infiltration, indicating that mild hypothermia can have neuroprotective effects by reducing the inflammatory reaction. PMID- 16258657 TI - Ullrich congenital muscular dystrophy and Bethlem myopathy: clinical and genetic heterogeneity. AB - Ullrich congenital muscular dystrophy (UCMD), due to mutations in the collagen VI genes, is an autosomal recessive form of CMD, commonly associated with distal joints hyperlaxity and severe course. A mild or moderate involvement can be occasionally observed. OBJECTIVE: To evaluate the clinical picture of CMD patients with Ullrich phenotype who presented decreased or absent collagen VI immunoreactivity on muscular biopsy. RESULTS: Among 60 patients with CMD, two had no expression of collagen V and their clinical involvement was essentially different: the first (3 years of follow-up) has mild motor difficulty; the second (8 years of follow-up) never acquired walking and depends on ventilatory support. A molecular study, performed by Pan et al. at the Thomas Jefferson University, demonstrated in the first a known mutation of Bethlem myopathy in COL6A1 and in the second the first dominantly acting mutation in UCMD and the first in COL6A1, previously associated only to Bethlem myopathy, with benign course and dominant inheritance. CONCLUSION: Bethlem myopathy should be considered in the differential diagnosis of UCMD, even in patients without fingers contractures; overlap between Ullrich and Bethlem phenotypes can be supposed. PMID- 16258658 TI - Dystrophin-glycoproteins associated in congenital muscular dystrophy: immunohistochemical analysis of 59 Brazilian cases. AB - The congenital muscular dystrophies (CMD) are heterogeneous muscular diseases with early and dystrophic pattern on muscle biopsy. Many different subtypes have been genetically identified and most phenotypes not yet identified belong to the merosin-positive (MP) CMD subgroup. OBJECTIVE: To analyze the immunohistochemical expression of the main proteins of the dystrophin-glycoproteins associated complex in muscle biopsy of patients with different CMD phenotypes, for investigating a possible correlation with clinical and histopathological data. METHOD: Fifty-nine patients with CMD had clinical, histopathological and immunohistochemical data evaluated: 32 had MP-CMD, 23 CMD with merosin deficiency (MD-CMD), one Ullrich phenotype and three Walker-Warburg disease. RESULTS: Dystrophin and dysferlin were normal in all; among the patients with MD-CMD, merosin deficiency was partial in nine who showed the same clinical severity as those with total deficiency; the reduced expression of alpha-sarcoglycan (SG) and alpha-dystroglycan (DG) showed statistically significant correlation with severe MD-CMD phenotype. CONCLUSION: There is a greater relationship between merosin and the former proteins; among MP-CMD patients, no remarkable immunohistochemical/phenotypical correlations were found, although the reduced expression of beta-DG had showed statistically significant correlation with severe phenotype and marked fibrosis on muscular biopsy. PMID- 16258659 TI - [Microanatomical study of the choroidal fissure in ventricular and cisternal approaches]. AB - The choroidal fissure is a narrow cleft in the medial part of the lateral ventricle, in a C-shaped arc, between the fornix and the thalamus, where the choroidal plexus join. Due to absence nervous tissue between ependyma and pia mater along this invagination, it is an important route in brain ventricles and cisterns. Five brains were studied by injecting colored silicone arteries and veins and five brains without colored silicone, in a total number of 20 brain hemispheres. It was analyzed and revised the neural, arterial and venous relationships and surgical approaches in all parts of the choroidal fissure. In conclusion, the previous knowledge detailed this microanatomy is primordial for neurosurgeons that will approach brain ventricular and cisternal lesions because the neurosurgeons gain a tridimensional notion that will be indispensable during surgery. PMID- 16258660 TI - [Cerebrovascular disease: language acquisition in preschool children]. AB - We describe ten children, aging 5 years and 1 month until 5 years and 11 months, when the phonoaudiological assessment was conducted. They are divided according to cerebrovascular disease, in CVD group (CVD-G) and control group (cG). Children were seen and CVD was confirmed in the acute phase at UNICAMP hospital. Audiologic assessment, protocol for Infant language assessment, and Peabody picture vocabulary test were used in the evaluations. The qualitative analysis of the subjects from a phonoaudiological and neurological point of view has shown the recovery of acquired language disorder (ALD) with no influence whatsoever in the development of 2 subjects and subtle language and/or learning process alterations for 3 subjects. The cases study has revealed that all aspects of language development in preschool children should be analyzed in an individual, quantitative, and qualitative basis to lead to conclusive findings. PMID- 16258661 TI - [Ultrasonic findings of intracranial hemorrhage in preterm neonates]. AB - Intracranial hemorrhage ICH is one of the most common neurological events in pre term newborn ICH is associated with low birth weight (< 1500 g) and gestational age (GA) at delivery (< 32 weeks). The most common site affected is the germinal matrix. Papile et al. classifies it at four grades. We analyzed, prospectively, 50 newborns (27 boys) with ultrasound diagnostic of ICH; all of them were pre term (GA < 37 weeks). They were classified according to sex, gestational age, birth weight and degree of ICH. The children were divided into two groups: A--GA < or = 33 weeks and B--34-37 weeks. In group A there were 34 children (25 boys) with mean GA of 31 weeks and birth weights average of 1308 g. In group B there were 16 children (2 boys), mean GA 34 weeks and birth weight average of 1951 g. The grades of ICH were: Group A--I-14, II-14, III-4 and IV-2; Group B--I-12, II-3 and III-1. The complications were more common in group A with 12 than group B with 4 children. The lesions happen in greatest number and most severity in children with low birth weight and younger (low gestational age). Ultrasound has shown to be effective for diagnostic and follow up of those children. PMID- 16258662 TI - [Transient cognitive impairment during generalized or diffuse epileptiform EEG discharges]. AB - Epileptiform EEG discharges (EED) are a frequent finding in people with epilepsy, and may occur in other clinical conditions. Formal testing during EEG recording may demonstrate transitory cognitive impairment (TCI). Thirty patients (27 with epilepsy) with generalized or diffuse EED on a previous examination were studied. The interaction between EED and cognitive tasks was evaluated on subsequent EEG. The numbers of correct or incorrect answers were compared with the occurrence or absence of EED. Results were analyzed by non parametric tests. TCI was characterized in 43.3% of the patients. In 23.3% the EED were rare and it was not possible to evaluate the occurrence of TCI. The findings confirm that otherwise subclinical EED may be accompanied by TCI. There is a need of research to evaluate the relevance of TCI and to determine when EED should be treated. PMID- 16258663 TI - [The initial testing and the discrimination property of the UFMG Sydenham's Chorea Rating Scale (USCRS)]. AB - Recently we developed and validated the Universidade Federal de Minas Gerais (UFMG) Sydenham's chorea Rating Scale (USCRS) to systematically assess SC patients. In this study, we assessed 97 children and adults with SC (mean age +/- SD, 15.5 +/- 5.9; male/female, 31/66) seen at the Movement Disorders Clinic at UFMG employing the USCRS. The patients were divided into 4 groups according to their clinical status: acute (n=19), recurrent (n=17), persistent (n=19) and remission (n=42). The mean +/- SEM USCRS scores for each group were: 47.7 +/- 4.7 for acute group, 29.5 +/- 2.6 for recurrent group, 17.6 +/- 3.1 for persistent group and 1.1 +/- 0.2 for remission group. All pair comparisons were statistically significant (p<0.05). Our results indicate that the USRSC can reasonably discriminate groups of SC patients in different clinical stages of the disease. PMID- 16258664 TI - [Neurological findings in a group of children and adolescents exposed and infected by HIV-1]. AB - The CNS infection by HIV-1 in infancy could be present immediately after infection or became manifest later. Microcephalia, mental retardation, pyramidal signs, humor and behavioral disorders and antiretroviral therapy complications are common. This is an observational, sectional and descriptive study about findings on neurological examination of 173 patients in a group of children and adolescents infected and exposed to HIV-1 in perinatal period. Most of them had more than one neurological finding or different diagnosis. The more common findings were: encephalopathy, mental retardation, language delay, pyramidal signs, hyporeflexia. The neurological examination was abnormal in 67% of all patients even in seroreverters. We suggest that this group has a high risk to neurological disease and the development of co-morbidity is directly correlated to clinical deterioration by HIV-1 infection. PMID- 16258665 TI - [Primary intracranial germ cell tumors in children: evaluation of fourteen cases]. AB - This study evaluates the diagnosis, therapy and survival of 14 patients with primary intracranial germ cell tumors during the period from 1991 to 2001. There were 11 males and 3 females. Mean age was 12.2 years old (20 days-18 years). On admission, the most common symptoms were headache (10/14), vomiting (6/14) and visual (6/14). The tumor was in pineal and hypothalamic region in 10 cases, suprasellar in 3 cases, and in the cerebral parenchyma in 1 case. Histologically there were 1 embryonal carcinoma, 5 germinomas, 2 mature teratomas, 1 immature teratoma and 5 mixed germ cell tumors. Treatment differed among the patients according to the type of tumor. Three patients died after tumor progression or relapse and one patient died from another condition. The remaining patients are alive and without disease. PMID- 16258666 TI - [Signs and symptoms associated to otoneurologic alterations diagnosed on computerized vestibular exam of patients with multiple sclerosis]. AB - OBJECTIVE: To identify main symptoms and signs on computerized vestibular testing in patients diagnosed with multiple sclerosis. METHOD: Thirty patients with the diagnosis of multiple sclerosis were studied. We analyzed data related to presented symptoms and the findings from a computerized vestibular testing realized in the otoneurological ambulatory in Santa Casa de Misericordia de Sao Paulo in 2003. RESULTS: Reported symptoms consisted mainly of disequilibrium (60%), tingling of limbs (43.3%), vertigo (40%), headache and anxiety (36.7%), tinnitus (30%), depression (26.7%). In vestibular testing we found alterations in positional nystagmus (6.7%), spontaneous nystagmus with the eyes shut (30%), directional nystagmus (13.3%) and caloric testing (63.3%). In the final conclusion of the exam, the diagnosis of irritative type peripheral vestibular syndrome (60%) and central syndrome (13.4%) prevailed. CONCLUSION: We conclude that the realization of an otoneurological exam has become indispensable in patients with multiple sclerosis. This is due to high prevalence of alterations at computerized vestibular exam and of otoneurological symptoms. PMID- 16258667 TI - [Somatosensory and motor evoked potentials in patients with cervical spondylosis]. AB - This study investigated 54 patients with cervical spondylosis (CS) with or without symptoms caused by cervical spondylotic myelopathy. Cervical MRI, somatosensory evoked potentials (SEPs) and motor evoked potentials (MEP) from upper and lower limbs of all of these patients were examined retrospectively. Were MRI findings the gold standard and the patients were classified in three groups. Group 1 (absence of spinal cord compression); Group 2 (presence of spinal cord indentation); Group 3 (spinal cord compression with alteration of intraspinal sign). The sensitivity of SEP of four limbs was 61.9%, the same one as the SEP of lower limbs. The sensitivity of MEP of four limbs was 71.4%, of the upper limbs alone was 66.7% and of the lower limbs alone 52.4%, thus showing the importance of using this method in all four limbs when cervical spondylotic myelopathy is suspected. The results shown by the study of the tibial nerve in SEP and of the abductor digiti minimi muscle in MEP were more frequently abnormals than the results shown by the study of the median nerve and extensor digitorum brevis muscle in the three groups, suggesting that exists a onset compromising of them. PMID- 16258668 TI - [Muscle activity during gait following stroke]. AB - OBJECTIVE: To compare muscle activity and joint moments in the lower extremities during walking between subjects with stroke and control subjects. METHOD: We compared fifteen healthy volunteers and fifteen stroke patients, with the same age gender and weight data had been compared by electromyography. The system of signals acquisition used consisted of five pairs of electrodes of surface, beyond one electrogoniometer on the axis articulate of rotation of the joint of the ankle in study. RESULTS: Onset times with respect to heel-strike for the medial gluteus, tibialis anterior, soleus, rectus femoris and medial hamstring muscles were significantly earlier during the gait cycle in subjects with stroke than in control subjects. The cessation times of soleus, tibialis anterior, rectus femoris, and medial hamstring muscles were significantly prolonged in subjects with stroke. CONCLUSION: Subjects with stroke showed more co-contractions of agonist and antagonist muscles at the ankle and knee joints during stance phase. These gait changes and co-contractions may allow subjects with stroke to adopt a safer, more stable gait pattern to compensate for diminished sensory information from the ankle. PMID- 16258669 TI - Tuberculosis: an uncommon cause of cerebral venous thrombosis? AB - Several infectious etiologies are related to cerebral venous thrombosis (CVT), but a review of literature showed only few cases related to tuberculosis (TB), and only one with neurological manifestations. We report an unusual case of CVT related to TB and mutation in prothrombin gene. A 38-man black presented abrupt right hemiparestesis, and hemiparesis. Investigations revealed CVT. Cerebral spinal fluid (CSF) examination evidenced an infection by Mycobacterium. He was heterozygous for G20210A prothrombin mutation. Probably, hypercoagulability mechanisms of TB, added to mutation of prothrombin gene increase the risk of CVT. PMID- 16258670 TI - Ventral extradural spinal meningeal cyst causing cord compression: neurosurgical treatment. AB - Spinal extradural meningeal cysts are typically formed by a thin fibrotic membranous capsule, macroscopically similar that of an arachnoid membrane, filled by cerebro spinal fluid and related to a nerve root or to the posterior midline. Ventral location is extremely rare and when it occurs they usually cause spinal cord herniation through the ventral dural gap. A 61 year-old man who began with a two years long history of insidious tetraparesis, spasticity and hyperreflexia in lower extremities, and flaccid atrophy of upper limbs, without sensory manifestations, is presented. Investigation through magnetic resonance imaging demonstrated an extensive spinal ventral extradural cystic collection from C6 to T11. The lesion was approached through a laminectomy and a cyst-peritoneal shunt was introduced. The cyst reduced in size significantly and the patient is asymptomatic over a 48 months follow-up. This is the first reported case of a spontaneous ventral extradural spinal meningeal cyst causing cord compression. Cyst-peritoneal shunt was effective in the treatment of the case and it should be considered in cases in which complete resection of the cyst is made more difficult or risky by the need of more aggressive surgical maneuvers. PMID- 16258671 TI - Traumatic pseudoaneurysm of the superficial temporal artery: case report. AB - A 26 year-old woman suffered a blunt head injury on the left temporal area and developed an arteriovenous fistula with a pseudoaneurysm on the superficial temporal artery. These fistulas are rare and usually associated with pseudoaneurysms, most commonly in the frontal arterial branch. The aneurysms generally appear late after trauma and present as a pulsatile painfully growing mass in the temporal region, associated with fremitus and bruit. The diagnosis is made by angiography and surgery is a very effective treatment. PMID- 16258672 TI - Bilateral isodense epidural hematoma: case report. AB - We present a case of a severe head injured 23 year-old male patient. The initial CT scan disclosed bilateral epidural hematoma, isodense with the brain, thus being a pitfall in diagnosis. Brief case report, image and literature review are presented. PMID- 16258673 TI - Metastatic prolactinoma: case report with immunohistochemical assessment for p53 and Ki-67 antigens. AB - Pituitary carcinomas are rare neoplasms characterized by craniospinal and/or systemic metastases originated from the pituitary. Their histopathology is frequently indistinguishable from that of benign adenomas. The development of markers that better reflect their behavior is desirable. We present the case of a 47 year-old man with a prolactin-secreting macroadenoma who was submitted to surgeries, cranial radiation therapy, and bromocriptine treatment, but evolved to a fatal outcome after the disclosure of intracranial metastases. Tumor samples underwent p53 and Ki-67 immunohistochemical assessment. p53 was absent in all samples, a rare finding among pituitary carcinomas. Ki-67 proliferative index was 2.80% in the original tumor, 4.40% in the relapse, and 4.45% in the metastasis. The figure in the relapse is higher than the expected for a noninvasive adenoma. In conclusion, p53 staining is not positive in all pituitary carcinomas. A high Ki-67 proliferative index in a pituitary adenoma might indicate a more aggressive behavior. PMID- 16258674 TI - Retropulsion and vertigo in the Chiari malformation: case report. AB - We describe a rare case of a 30 year-old woman with intense vertiginous sensation, lack of body balance and a tendency to fall backwards, making it necessary for two people to sustain her. The magnetic resonance imaging of the craniocervical junction evidenced tonsilar herniation at the inferior level of C1, and during the operation performed in sitting position, we observed crowding of the cerebellar tonsils at the level of C3. After the osteo-dural-neural decompression, the symptomatology remitted on the same day of the operation. PMID- 16258675 TI - [Motor nerve conduction study in McArdle disease: case report]. AB - McArdle disease (glycogenosis type V) is a metabolic myopathy with symptoms of exercise intolerance caused by deficiency of the enzyme myophosphorylase. In these patients, the motor nerve conduction studies after a short period of maximal voluntary muscle contraction or repetitive stimulation reveals characteristic findings of the disease. A 37-year-old man presented symptoms of exercise intolerance, muscular fatigue and cramps in the beginning of the physical activity with "second wind" phenomenon. The motor nerve conduction studies after a voluntary contraction of 30 and 90 seconds presented decrease in the amplitude of the compound muscle action potential in median, ulnar and deep peroneal nerves; and decrement after 200 stimulation at the 40 Hz in deep peroneal nerve. The electromyography presented myopathic pattern and during the ischemic exercise electric silence was not proven. The characteristic of electrophysiological studies are discussed with emphasis at the importance of the motor nerve conduction studies in the patients with suspicion of metabolic myopathy. PMID- 16258676 TI - [Vascular Wallerian degeneration on magnetic resonance in a patient with probable progressive supranuclear palsy: aetiology or casual link?]. AB - Progressive supranuclear palsy (PSP) is one of the most important causes of parkinsonism non responsive to therapy. Vascular parkinsonism is not uncommon. However, the cause-effect relationship between them is uncertain. We report on a 65 year old man with probable PSP who developed the clinical features of the disease after a ischaemic stroke. Magnetic resonance imaging disclosed a corticospinal tract Wallerian degeneration. There is not such an observation in the literature about this possible correlation. PMID- 16258677 TI - [Fibrolipomatous hamartoma of the median nerve: case report]. AB - Fibrolipomatous hamartoma is a rare benign neoplasm that in some cases is associated with macrodactylia. We describe a 31-year-old man who had a tissue enlargement in the wrist, second and third fingers of the left hand since infancy. At 23-years-old he began with continuous, progressive and high intensity pain that occurred more frequently at night, localized in the left hand. It was associated with paraesthesias and hypostesias predominantly at the fingers described above. Investigation with X-ray, ultrasonography, electrodiagnosis, magnetic resonance image of the left wrist and hand showed carpal tunnel syndrome with macrodactylia by fibrolipomatous hamartoma of the median nerve. The patient did not a have good response to clinical therapy, so he was submitted to a surgical decompression of the left carpal tunnel, and after three months of follow up is asymptomatic. PMID- 16258678 TI - [Cranial idiopathic hypertrophic pachymeningitis associated with orbital pseudotumor: case report]. AB - Hypertrophic pachymeningitis is a rare disorder characterized by meningeal thickness, that can be caused by infection, tumoral infiltration, inflammatory disorders or idiopathic. We report the case of a 40 year-old man that presented with longstanding headache and progressive bilateral visual loss and proptosis. Cranial and orbital magnetic resonance imaging revealed diffuse dural thickness and bilateral extraconal orbital lesion. Extensive investigation did not reveal any systemic condition. Histopathological study after meningeal and orbital biopsy disclosed a chronic inflammatory process compatible respectively with idiopathic hypertrophic pachymeningitis (IHP) and orbital pseudotumor. This case emphasizes that orbital involvement can occur in IHP and that its early identification is of great importance in order to improve the visual prognosis of this condition. PMID- 16258679 TI - [Neuroprotective agents in stroke: national opinion]. AB - The Brazilian Stroke Society constituted a committee composed by specialists from different areas of Brazil that emitted a viewpoint called "National Opinion", written similar to the consensus pattern. The study purpose is to guide and offer subsidies for diagnosis and therapeutical plans for different situations in cerebrovascular diseases. The current article analyses "neuroprotective agents in stroke", discussing the level of evidence for the use of potential neuroprotective drugs and ongoing clinical trials. PMID- 16258680 TI - [Guidelines for the treatment of multiple sclerosis with immunomodulatory drugs]. AB - The treatment of patients with multiple sclerosis (MS) with immunomodulatory drugs, and more recently, with immunosuppressive drugs, have modified the natural history of the disease in the last years. The conclusions and recommendations elaborated by several authors based upon multicenter studies make us review and update concepts, proposing modifications to government institutions in order to improve the assistance to MS patients, the main purpose of this work. Herein, the Neuroimmunology Scientific Department of the Brazilian Academy of Neurology judged important to update the recommendations concerning MS diagnosis criteria, classification of progression patterns, foundation of reference centers and the use of immunomodulatory drugs. PMID- 16258683 TI - The IX International Meeting on Paracoccidioidomycosis, Aguas de Lindoia, SP, Brazil, October 2-5, 2005. Abstracts. PMID- 16258694 TI - The occurrence of acute postoperative confusion in patients after cardiac surgery. AB - This study quantified the occurrence of acute confusion in cardiac surgery patients at three German hospitals. A total of 867 patients, 22-91 years old, were examined each nursing shift postoperatively for 5 days for the presence of acute confusion using a modified version of the Glasgow Coma Scale and Confusion Rating Scale. The night shifts and the third postoperative day showed the most frequent periods of occurrence. Confusional state was noted in patients ranging from 10.5% for patients aged <70, to 40.7% for patients >80 years of age. Those found at increased risk were patients of increasing age and coexisting disease. Targeted nursing interventions for patients at increased risk of acute confusion may decrease this complication. PMID- 16258695 TI - Investigation of penetration abilities of various oils into human hair fibers. AB - In this work we have explored capillary adhesion between hair fibers treated with different types of oils. With coconut, olive, and sunflower oils the capillary adhesion was found to decrease with time, but not with mineral oil. Application of heat reduced the capillary adhesion further for coconut and sunflower oils. Again, this was not observed with mineral oil. Based on an earlier study, where coconut oil was found to penetrate hair while mineral oil was unable to do so, it was hypothesized that the reduction in capillary adhesion resulted from the penetration of oil into the fiber, leaving a thinner oil film on the surface. Such a reduction in capillary adhesion can be explained by changes in Laplace pressure and in the areas of liquid bridges formed between the fibers. The thinning of oil films on the surface of hair has been confirmed independently by goniophotometric measurements on single hair fibers treated with coconut, sunflower, and mineral oils. Thick films of oil (thicker than approximately 0.5 microm) are known to mask the scale structure. As the oil is absorbed into the hair, the film thins with time and application of heat, and the scale structure reappears. This change can be conveniently determined by measuring the scale angle, using the well established goniophotometric protocol. The agreement between the two methods supports the concept that the reduction in capillary adhesion between hair fibers is most likely due to thinning of oil films by absorption of oil into the hair. PMID- 16258696 TI - A novel method for visualizing hair lipids at the cell membrane complex: argon sputter etching/scanning electron microscopy. AB - Hair lipids localized at the cell membrane complex (CMC) play a part in chemical diffusion, cell cohesion, and mechanical strength. There is no method currently available to visualize hair lipids at the CMC. We found that scanning electron microscopy (SEM) of a transversely polished hair plane followed by argon sputter etching (ASE) provides a specific characteristic image consisting of circular patterns (CP) and stitch patterns (SP) at the cortex. Both the CP and the SP are formed as convex structures and are associated with melanin granules and the CMC, respectively. While the convex formation of the CP is not affected by any treatments tested, that of the SP disappeared following treatment of hair fibers with organic solvents and reappeared following incubation of the solvent-treated hair fibers with melting lipids, which suggests that the hair lipids are responsible for the convex SP. Other treatments, such as chemical fixation, thin sectioning, and pre-/post-incubation of the hair plane, reduce or abolish the convex formation of the SP. These findings suggest that the following pathway leads to the convex formation of SP during ASE: (a) joule heat is generated on the surface by violent collisions of argon ions, (b) melting CMC lipids ooze out from the inside to the surface, and (c) CMC lipids that have oozed out are chemically changed, leading to the final convex formation of the SP. With ASE SEM, visualization of hair lipids as convex structures of SP should enable us to characterize the fine structure and localization of hair lipids and to clarify the roles and functions of the CMC of human hair. PMID- 16258697 TI - Effect of systemic hormonal cyclicity on skin. AB - Fluctuations in estrogen and progesterone during the menstrual cycle can cause changes in body systems other than the reproductive system. We conducted several studies to determine a possible correlation between phases of the menstrual cycle and specific skin properties. Healthy Caucasian women (ages 21-48), who had a typical 26-29 day menstrual cycle, participated in the studies. Measurements of skin barrier strength, dryness, response to lactic acid stinging, skin surface lipids, and microflora were obtained every week for two to three months. Ultraviolet B susceptibility in terms of minimal erythemal dose was also studied. The skin barrier was the weakest between days 22 and 26 of the cycle. Elevated neuronal response (lactic acid sting) was not observed to vary much with the cycle. Skin was driest between day 1 and day 6, while skin surface lipid secretion appeared to be highest on days 16-20 of the hormonal cycle. The highest microbial count was around days 16-22, and there was a high UV-B susceptibility between days 20 and 28 of the menstrual cycle. PMID- 16258698 TI - Penetration of cationic conditioning compounds into hair fibers: a TOF-SIMS approach. AB - Cationic conditioning compounds protect against hair damage caused by cosmetic chemical treatments and grooming practices. They also enhance the retention of moisture. However, the question as to whether they do this superficially by residing on the hair surface or by penetrating into the fiber remains unanswered. In this work, an attempt has been made to show the penetration of a low-molecular weight cationic conditioning compound into the hair cortex using the time-of flight secondary ion mass spectrometry (TOF SIMS) method, applied in earlier research to show the penetration of oils into hair. An example of the practical benefit of such penetration into the cortex in greatly improving the fatigue resistance of hair has been discussed. PMID- 16258702 TI - N-cadherin expression in breast cancer: correlation with an aggressive histologic variant--invasive micropapillary carcinoma. AB - Upregulation of N-cadherin in epithelial tumor cells has been shown to contribute to the invasive/metastatic phenotype. It remains however to be determined whether N-cadherin is increased in human breast cancers with enhanced malignant potential. We examined a large number of invasive breast cancer specimens (n = 114) for N- and E-cadherin. These specimens compared invasive duct carcinomas (IDCs) of varying histologic grades with an aggressive subtype, invasive micropapillary carcinoma of the breast (MPAP), which has a high propensity for lymphatic invasion and lymph node metastasis. Staining scores for N- and E cadherin were compared between non-MPAP and MPAP IDCs, and between the invasive and ductal carcinoma in situ (DCIS) of each IDC using statistical analysis. We found that N-cadherin was expressed in 76% of MPAP and 52% of non-MPAP carcinomas, and E-cadherin in 57% of MPAP and 36% of non-MPAP tumors. More MPAP (25%) compared to non-MPAP (5%) tumors expressed both cadherins. Of the two cadherins, N-cadherin was significantly associated with MPAP tumors (p = 0.033) compared to E-cad (p = 0.171). Moreover, in the majority of tumors that were positive for N-cadherin, the staining scores were increased in the IDC relative to intraductal components, and this effect was more dramatic in the MPAP carcinomas. This difference for N-cadherin was greater than the corresponding difference for E-cadherin in the MPAP group (p = 0.005), whereas such changes were not significant in the non-MPAP group (p = 0.10). Thus, N-cadherin is associated with tumor aggressiveness and metastatic potential and may contribute to tumor progression. PMID- 16258703 TI - Adaptation of estrogen-regulated genes in long-term estradiol deprived MCF-7 breast cancer cells. AB - First line treatment of hormone dependent breast cancer initially causes tumor regression but later results in adaptive changes and tumor re-growth. Responses to second line treatments occur but tumors again begin to progress after a period of 12???18??months. In depth understanding of the adaptive process would allow the identification of targets to abrogate the development of hormonal resistance and prolong the efficacy of endocrine therapy. We have developed a model system to examine adaptive changes in human MCF-7 breast cancer cells. Upon deprivation of estradiol for a prolonged period of time, a maneuver analogous to surgical oophorectomy in pre-menopausal women and use of aromatase inhibitors in post menopausal patients, tumor cells adapt and become hypersensitive to estradiol. We reasoned that the expression pattern of multiple genes would change in response to estradiol deprivation and that cDNA microarrays would provide an efficient means of assessing these changes. Accordingly, we examined the transcriptional responses to estradiol in long-term estradiol deprived (LTED) MCF-7 cells with a cDNA microarray containing 1901 known genes and ESTs. To assess the changes induced by long-term estradiol deprivation, we compared the effects of estradiol administration in LTED cells with those in MCF-7 cells, which we had previously reported, and confirmed with real time PCR using the parental and LTED cells. Seven genes and one EST were induced by estradiol in LTED but not in wild type MCF-7 cells, whereas ten genes were down-regulated by estradiol only in LTED cells. The expression of seven genes increased concurrently and five decreased in response to estradiol in both cell types. From these observations, we generated testable hypotheses regarding several genes including DKFZP, RAP-1, ribosomal protein S6, and TM4SF1. Based upon the known functions of these genes and the patterns of observed changes, we postulate that divergent regulation of these genes may contribute to the different biologic responses to estrogen in these cell lines. These results provide targets for further mechanistic studies in our experimental system. Our findings indicate that long-term estradiol deprivation causes expression changes in multiple genes and emphasizes the complexity of the process of cellular adaptation. PMID- 16258704 TI - Conservative treatment of breast cancer: its evolution. AB - BACKGROUND: Over recent decades, breast carcinoma surgery has witnessed a considerable evolution. The extent of surgery undertaken has progressively reduced, leading to less disfigurement and a significant improvement in quality of life, thereby offering women considerable motivation to seek early diagnosis. From an oncological perspective, outcome was found to be equally effective, and this key observation provided us with significant impetus to investigate the effects of reducing the radiotherapy field. PATIENTS AND METHODS: We present the achievements made so far in the conservative treatment of breast carcinoma, based on experiences at the European Institute of Oncology in Milan and, prior to 1994, at the Milan Cancer Institute. RESULTS AND DISCUSSION: Conservative surgery of breast carcinoma, both in the breast and in the axillary nodes, has yielded very good results in terms of overall survival and impact on quality of life. The reduction of the radiation field, conducted in our Institute by means of electron intraoperative radiotherapy (ELIOT), is currently the subject of evaluation via a randomised trial. PMID- 16258706 TI - Immunogenic hsp-70 is overexpressed in colorectal cancers with high-degree microsatellite instability. AB - PURPOSE: Colorectal cancers that display high-degree mi-crosatellite instability are associated with an improved prognosis and evidence of an activated host immune response. Molecular analyses have suggested that heat shock proteins, a family of proteins that have key immunologic functions, are upregulated in these cancers. We aimed to explore the expression of heat shock proteins 70 and 110 and their relationship to microsatellite instability, survival, and other clinicopathologic parameters. METHODS: Twenty-six colorectal cancers that displayed microsatellite instability were matched by age, stage, and site in the colorectum to 26 microsatellite-stable cancers. Immunohistochemistry was used to detect expression of both markers. RESULTS: The microsatellite-unstable group showed significantly higher expression of heat shock protein 70 than the microsatellite-stable group (P = 0.006), and patients undergoing curative resections for unstable cancers had improved prognosis compared with their stable counterparts (P = 0.026). Significantly, in a multivariate survival analysis, low or absent heat shock protein 70 expression was independently associated with a poor outcome (P = 0.001). CONCLUSIONS: Heat shock protein 70 has known functions that promote antitumor immune responses. Its overexpression in colorectal cancers with microsatellite instability may be pivotal to explaining these tumors' enhanced immunogenicity and improved prognosis. PMID- 16258705 TI - Mesh migration following repair of inguinal hernia: a case report and review of literature. AB - New surgical procedures in the aftermath of their benefits beget new complications as well. Laparoscopic repair of inguinal hernia is widely practised now. It involves reinforcement of hernial defect with a synthetic mesh. We report a rare case of migration of mesh into urinary bladder following this mode of hernial repair. We also present a review of literature involving migration of mesh following all inguinal hernial repairs. PMID- 16258707 TI - Phlebosclerotic colitis with deep circumferential ulceration: three-year endoscopic follow-up. Report of a case. AB - We report a case of phlebosclerotic colitis with deep circumferential ulceration in which the characteristic findings were observed radiologically and endoscopically. Previously, the patient was diagnosed with colitis of unknown etiology in 1999 when a colonoscopy showed small erosions, ulcers, and dark purple mucosa in the right colon. As a result of parenteral nutrition treatment, his symptoms and ulcerations disappeared; however, the dark purple mucosa remained unchanged for three years on the annual endoscopic follow-up. Recurrent colitis associated with circumferential ulceration was diagnosed in 2002. The patient was treated again with only parenteral nutrition and his symptoms improved after a few days; the ulceration completely disappeared within seven months. This case emphasizes the benefit of nonsurgical treatment for phlebosclerotic colitis even in cases of deep circumferential ulceration. PMID- 16258708 TI - Prospective, multicenter evaluation of highly concentrated fibrin glue in the treatment of complex cryptogenic perianal fistulas. AB - PURPOSE: The surgical management of complex perianal fistulas is challenging and may be associated with the risk of sphincter injury. Instillation of fibrin glue to the fistula tract is a simple procedure that does not involve any muscle division, and potentially results in healing of the fistula. This study was designed to assess the use of highly concentrated fibrin glue with intra-adhesive antibiotics in the treatment of complex cryptogenic perianal fistulas. METHODS: Patients with complex perianal fistulas of cryptogenic origin were prospectively included in this multicenter study. Injection of the fibrin glue mixed with antibiotics was performed in a uniform fashion. After the procedure, patients were actively examined at fixed time intervals; in cases of recurrent fistula, reinjection of fibrin glue was offered. RESULTS: Sixty patients were enrolled; complete healing of the fistula was achieved in 32 patients (53 percent). Eight of 28 patients (29 percent) who were not completely healed had significant symptomatic improvement. All patients resumed normal daily activity the day after surgery and none had any deterioration in continence related to the procedure. The majority of the 26 (43 percent) adverse events were considered mild and spontaneously resolved; 2 patients (3 percent) with perianal septic complications were successfully treated by drainage. CONCLUSIONS: Injection of fibrin glue for the treatment of perianal fistulas is safe, simple, and associated with early return to normal activity. Although moderately successful, it may preclude extensive surgery in more than one-half of these patients. PMID- 16258709 TI - Telephone vs. face-to-face biofeedback for fecal incontinence: comparison of two techniques in 239 patients. AB - PURPOSE: Biofeedback is an effective treatment for patients with fecal incontinence, yet little is known about how it works or the minimum regime necessary to provide clinical benefit. This study compares the effectiveness of a novel protocol of telephone-assisted biofeedback treatment for patients living in rural and remote areas with the standard face-to-face protocol for patients with fecal incontinence. METHODS: A new treatment program comprising an initial face to-face assessment and treatment with transanal manometry and ultrasound biofeedback, followed by three treatments conducted via telephone and a final face-to-face assessment, was developed. Standard treatment involved five face-to face treatment sessions with manometry and ultrasound. Patients from rural areas were offered the telephone-assisted treatment protocol. Data gathered prospectively included incontinence scores, a quality of life index, anal manometry, and external sphincter isometric and isotonic fatigue times. RESULTS: A total of 239 consecutive patients treated between July 2001 and July 2004 were enrolled. There were no significant differences in demographic details, past history, or pretreatment measures of the two groups. Forty-six of 55 patients (84 percent) treated with the telephone protocol and 129 of 184 (70 percent) treated by the standard technique completed treatment. There were substantial, significant improvements after treatment, including 54 percent mean improvement in patient's own rating of their incontinence in both groups; a mean decrease of 3.1 and 3.2 on the St. Mark's incontinence score (from 7.9 to 4.7 and 7.4 to 4.2 of 13) and relative improvements of 128 and 130 percent in the quality of life index (from 0.29 to 0.65 and 0.3 to 0.69 of 1) for the telephone-assisted and standard groups respectively. Importantly, there were no significant differences between the telephone-assisted or standard groups in any outcome. Of patients who completed treatment, 78 percent were better or much better. CONCLUSIONS: A less intensive regime of biofeedback seems to be equally effective as the standard intensive protocol. This finding adds weight to the evolving concept that the physical aspects of biofeedback treatment, such as manometry or ultrasound, may not be necessary in the treatment of most patients with fecal incontinence. This needs to be further tested in a randomized, controlled trial. PMID- 16258710 TI - Arterial thromboembolic complications of inflammatory bowel disease. PMID- 16258711 TI - Malignancies of the appendix: beyond case series reports. AB - PURPOSE: A comprehensive analysis was performed for five histologic types of appendiceal tumors to compare incidence, clinicopathologic features, survival, and appropriateness of surgery. METHODS: All patients diagnosed with mucinous adenocarcinoma (n = 951), adenocarcinoma (n = 646), carcinoid (n = 435), goblet (n = 369), and signet-ring cell (n = 113) in the Surveillance, Epidemiology, and End Results database (1973-2001) were analyzed. Evaluation of incidence, stage, and five-year relative survival were determined for each histology. The appropriateness of the operative procedure (i.e. , appendectomy vs. colectomy) was examined by tumor type and size. RESULTS: Tumor incidence, patient demographics, survival outcomes, and appropriateness of surgery varied significantly among the different appendiceal tumor histologies. The most common appendiceal tumors were mucinous. With regard to patient demographics, carcinoids presented at an earlier mean age of 41 years and 71 percent were female (P < 0.001 for both). Overall five-year survival was highest for carcinoid (83 percent) and lowest for signet ring (18 percent). Although current guidelines specify that a right hemicolectomy (rather than an appendectomy) be performed for all noncarcinoid tumors and carcinoid tumors >2 cm, we found that 30 percent of noncarcinoids underwent appendectomy. Similarly, 28 percent of carcinoids >2 cm under-went appendectomy, which is a lesser resection than is indicated. CONCLUSIONS: This study provides a population-based analysis of epidemiology, tumor characteristics, survival, and quality of care for appendiceal carcinomas. This characterization provides a novel description of the presentation and outcomes for malignancies of the appendix and highlights that a substantial number of patients with appendiceal tumors may not be receiving appropriate surgical resection. PMID- 16258712 TI - Practice parameters for the surgical treatment of ulcerative colitis. AB - The American Society of Colon and Rectal Surgeons is dedicated to assuring high quality patient care by advancing the science, prevention, and management of disorders and diseases of the colon, rectum, and anus. The Standards Committee is composed of Society members who are chosen because they have demonstrated expertise in the specialty of colon and rectal surgery. This committee was created to lead international efforts in defining quality care for conditions related to the colon, rectum, and anus. This is accompanied by developing Clinical Practice Guidelines based on the best available evidence. These guidelines are inclusive, and not prescriptive. Their purpose is to provide information on which decisions can be made, rather than dictate a specific form of treatment. These guidelines are intended for the use of all practitioners, health care workers, and patients who desire information about the management of the conditions addressed by the topics covered in these guidelines. It should be recognized that these guidelines should not be deemed inclusive of all proper methods of care or exclusive of methods of care reasonably directed to obtaining the same results. The ultimate judgment regarding the propriety of any specific procedure must be made by the physician in light of all of the circumstances presented by the individual patient. PMID- 16258713 TI - 1,4-Dimethoxybenzene, a floral scent compound in willows that attracts an oligolectic bee. AB - Many bees are oligolectic and collect pollen for their larvae only from one particular plant family or genus. Here, we identified flower scent compounds of two Salix species important for the attraction of the oligolectic bee Andrena vaga, which collects pollen only from Salix. Flower scent was collected by using dynamic-headspace methods from Salix caprea and S. atrocinerea, and the samples were subsequently analyzed by coupled gas chromatographic-electroantennographic detection (GC-EAD) to detect possible attractants of A. vaga. EAD active compounds were identified by gas chromatography coupled to mass spectrometry. Both Salix species had relatively similar scent profiles, and the antennae of male and female bees responded to at least 16 compounds, among them different benzenoids as well as oxygenated monoterpenoids and sesquiterpenoids. The strongest antennal responses were triggered by 1,4-dimethoxybenzene, and in field bioassays, this benzenoid attracted females of A. vaga at the beginning of its flight period, but not at the end. PMID- 16258715 TI - Cardiac remodeling as therapeutic target: treating heart failure with Cardiac Support Devices. PMID- 16258714 TI - Disability and the detection of mental disorder in primary care. AB - INTRODUCTION: Despite the importance of disability associated with common mental disorders, research on the detection of mental disorders in primary care has scarcely explored its relevance. Aims To describe the disability burden of primary care patients with common mental disorders and subthreshold disorders and to examine the association between general practitioner's (GP) recognition of mental disorder and disability. Design Cross-sectional survey of GPs and their patients. Setting General practices in the lower North Island of New Zealand. Method Participants were randomly selected: GPs (n=70) and their patients (n=3414, of whom a subset of 775 from the basis of this paper). Formal DSM-IV diagnoses were made with the Composite International Diagnostic Interview (CIDI), and psychosomatic and psychological symptoms were measured with the Somatic and Psychological Health Report. Disability was measured with the World Health Organisation's Disability Assessment Schedule-version II. GPs independently rated the severity of psychological symptoms and the presence or absence of disorder. Results The principal findings were (1) that disability was associated with both mental disorder and subthreshold disorder with no significant difference in the level of disability between these categories, and (2) that GPs were less sensitive to the presence of mental disorders as defined by the CIDI if there was little concomitant disability, and in subthreshold cases, the presence of disability increased the chance of GPs identifying clinically significant symptoms. Conclusion Studies of GP recognition of mental disorder have almost exclusively adopted the perspective of concepts of disorder as defined by psychiatry. This study provides some insight into the way GPs attend to both symptoms and functioning in their assessments of psychological syndromes. Disability is an important cue to recognition of mental health problems in the primary care setting, including those that are not recognised by standardised psychiatric assessment but which may still be relevant to patient suffering. PMID- 16258716 TI - Left ventricular size and shape: determinants of mechanical signal transduction pathways. AB - Heart failure may be viewed as a progressive disorder that is impelled, at least in part, by progressive left ventricular (LV) remodeling. In the present discussion we will review the role of LV remodeling in the pathogenesis of heart failure, with a focus on the contribution that changes in LV size and shape play in the development and progression of the progression of heart failure. The clinical implication of this review is that existing neurohormonal strategies may not completely prevent disease progression in the failing heart, and that adjunctive strategies that are designed to specifically prevent and/or attenuate LV remodeling may play an important role in the clinical treatment of heart failure. PMID- 16258717 TI - Design and features of the Acorn CorCap Cardiac Support Device: the concept of passive mechanical diastolic support. AB - The Laplace equation points to the importance of ventricular wall stress as a factor in heart failure development and progression. Based on animal and clinical experience with active and passive cardiomyoplasty, a synthetic passive constraint was proposed as a means of reducing excessive wall stress, and thus assuaging disease progression. The Acorn CorCap CSD Cardiac Support Device was designed to provide passive diastolic support and serve as a constraint against chronic cardiac dilation in heart failure. Basic and uncomplicated in appearance, the device nevertheless incorporates numerous sophisticated structural and functional features intended maximize therapeutic value. The device is fabricated from medical grade multifilament polyester yarn, in a specific knit construction chosen to provide structural integrity, low physical profile, pliability for intimate contact with the epicardial surface, ease of manipulation during implantation, and anisotropic compliance characteristics to encourage beneficial reverse remodeling. PMID- 16258718 TI - Global left ventricular remodeling with the Acorn Cardiac Support Device: hemodynamic and angiographic findings in dogs with heart failure. AB - Preventing progressive left ventricular (LV) remodeling is paramount in the treatment of heart failure. In recent years, several surgical approaches have been implemented with the objective of improving LV function through amelioration of progressive LV remodeling. These included surgical reduction of LV size, the so-called Batista procedure, dynamic cardiomyoplasty and mitral valve repair to limit or eliminate functional mitral regurgitation. While the Batista procedure and dynamic cardiomyoplasty have for all practical purposes been abandoned, the lessons learned from these procedures gave rise to a new generation of devices aimed at preventing progressive LV dilation and restoring LV shape by passive mechanical containment of the failing LV. One such device is the Acorn Cardiac Support Device (CSD) or the CorCap. Studies in dogs with intracoronary microembolization-induced moderate and advanced heart failure have shown that long-term monotherapy with the CSD not only prevents progressive LV dilation but, in effect, partially reverses this phenotype. These studies have also shown that the CSD restores, albeit in part, progressive LV chamber sphericity and attenuates functional mitral regurgitation. These benefits were accompanied by improvement in global LV function along with improvements of remodeling at the cellular level. The findings were largely responsible for initiating safety and feasibility clinical trials with the CSD and ultimately, the initiation of the Acorn efficacy trial that was completed in 2004. This review will focus on studies conducted in dogs with heart failure and, specifically on hemodynamic, angiographic and echocardiographic results from these studies that provided support for the CSD as a successful technology targeting "reverse LV remodeling" for the treatment of heart failure. PMID- 16258719 TI - Efficacy of the Acorn Cardiac Support Device in animals with heart failure secondary to high rate pacing. AB - AIM: To utilise an ovine model of tachycardia induced progressive dilated cardiomyopathy and heart failure to investigate the efficacy of passive ventricular constraint with the Acorn cardiac support device as a heart failure treatment. METHODS: (a) Moderate heart failure was produced in 16 sheep by pacing for 3 weeks. Half were implanted and half sham implanted with the CSD. Pacing continued at a higher rate for an additional 3 weeks. Cardiac function was assessed by echocardiography and manometry. (b) Moderate heart failure was produced (as above) in 27 sheep, 9 were implanted with CSD, pacing was restarted for 4 weeks, the initial CSD implants were terminated and another 9 animals were CSD implanted (severe heart failure), pacing was restarted in the remaining 18 animals for an additional 4 weeks (total 12 weeks) and then all animals were terminated. Cardiac function was assessed using echocardiography and treadmill exercise testing. RESULTS: (a) After 6 weeks of rapid pacing CSD implant animals had significantly better cardiac function both when compared with pre implant values and with non-implanted animals at termination. (b) CSD implantation at both moderate and severe failure resulted in significant improvements in cardiac function both when compared with pre implant values and with non-implanted animals at termination. When compared to pre implant values the improvement was greatest in severe implant animals. CONCLUSION: In this ovine model of tachycardia induced progressive heart failure, CSD implantation in either moderate or severe heart failure resulted in improved cardiac function, reduced left ventricular volume and mitral regurgitation both when compared with function at time of implant and with non implanted control animals. PMID- 16258720 TI - Infarct size reduction and attenuation of global left ventricular remodeling with the CorCap cardiac support device following acute myocardial infarction in sheep. AB - BACKGROUND: Whether mechanical restraint of the left ventricle (LV) can influence remodeling following myocardial infarction (MI) remains poorly understood. The following discussion details three studies examining the effects of surgically placing a cardiac support device (CSD) over the entire epicardial surface, on infarct expansion, global cardiac function and myocyte geometry and function post MI. METHODS: The effects of passive constraint on infarct expansion and global cardiac function/myocardial energetics were investigated in 10 sheep (5 MI only; 5 MI + CSD) using pressure-volume analysis and magnetic resonance imaging (MRI). Additionally, 11 sheep (5 MI only; 6 MI + CSD) were used to study the effects of passive restraint on myocyte geometry and function post-MI, with 10 additional uninstrumented sheep serving as controls. Baseline data was collected followed by the creation of an anterior infarct. 1 week post-infarct the animals underwent a second set of data collection studies followed by placement of the CSD in the experimental groups. Additional data was collected at 2 and 3 months post-MI. The animals in the myocyte function group underwent additional studies immediately following the 3 month time point. RESULTS: Infarct expansion was diminished as a result of the CSD. At 1 week post-MI the akinetic area was similar in both groups. At the terminal time-point, the akinetic area in the control group was similar to the 1-week time-point whereas, in the CSD group, the area of akinesis decreased (P = 0.001). A comparison of the two groups at the terminal time-point demonstrates a significantly diminished area of akinesis in the CSD group (P = 0.004). The relative area of akinesis followed a similar pattern. The CSD group also exhibited a decrease in end-diastolic volume (control 110.3 +/- 19.8 mL vs. CSD 67.6 +/- 4.7 mL, P = .006) and an improved ejection fraction (control 15.5% +/- 5.7% vs. CSD 29.46% +/- 4.42%, P = .008) relative to the control group. Myocardial energetics were also enhanced in the CSD group as evidenced by significant improvements in potential energy (control 2,015 +/- 503 mL x mm Hg/beat vs. CSD 885 +/- 220 mL x Hg/beat, P = .006), efficiency (control 39.4% +/ 13.6% vs. CSD 59.8% +/- 8.5%, P = .044), and oxygen consumption (control 0.072 +/- 0.013 mL O(2)/beat vs. CSD 0.052 +/- 0.007 mL O(2)/beat, P = .034). Isolated LV myocyte shortening velocity was reduced by 35% from control values (P < 0.05) in both MI groups. LV myocyte beta-adrenergic response was reduced with MI, but normalized in the MI + CSD group. Relative collagen content was increased and matrix metalloproteinase-9 was decreased within the MI border region of the CSD group. CONCLUSIONS: The CorCap cardiac support device retarded infarct expansion, improved global and regional cardiac function and beneficially modified LV and myocyte remodeling post-MI. These findings provide evidence that non pharmacological strategies can interrupt adverse LV remodeling post-MI. PMID- 16258721 TI - Left ventricular histomorphometric findings in dogs with heart failure treated with the Acorn Cardiac Support Device. AB - Progressive left ventricular (LV) dilation in the setting of heart failure is associated with increased mortality and morbidity. The Acorn Cardiac Support Device (CSD, Acorn Cardiovascular, Inc., St. Paul, MN) is a preformed polyester device that is surgically placed over the cardiac ventricles, anchored to the AV groove and tailored anteriorly to fit snugly over the epicardial surface of the heart. The CSD was shown to prevent progressive LV enlargement and, indeed, reduce LV size and attenuate global LV remodeling in both animal models of experimentally-induced heart failure as well as in patients with advanced heart failure. This review will examine the CSD from two histologic perspectives namely, (1) the interaction of the CSD with the epicardial surface of the heart and (2) the effects of long-term therapy with the CSD on cellular remodeling. The review will be based on available pre-clinical data generated in dogs with coronary microembolization-induced heart failure that underwent long-term (3 and 6 months) monotherapy with the CSD. The data will show that long-term implantation leads to encapsulation of the CSD by connective tissue that matures with time and that does not invade the underlying myocardium. Furthermore that implantation of the CSD has no adverse impact on epicardial coronary vessel. At the cellular level, existing data will show that long-term monotherapy with the CSD is associated with reduced cardiomyocyte hypertrophy, reduced volume fraction of replacement and interstitial fibrosis, normalization of capillary density and oxygen diffusion distance and attenuation of cardiomyocyte apoptosis. The outcomes strongly argue in favor of a structural modification of the failing myocardium by CSD therapy that is consistent with "reverse cellular remodeling". PMID- 16258722 TI - Improvement of cardiac sarcoplasmic reticulum calcium cycling in dogs with heart failure following long-term therapy with the Acorn Cardiac Support Device. AB - Abnormal Ca(2+)-homeostasis is a hall-marked characteristic of the failing heart. In the normal myocardium, the sarcoplasmic reticulum (SR) is a principal organelle that controls intracellular Ca(2+) concentration during the cardiac cycle. The SR consists of longitudinal and terminal cisternea regions. The former contains the Ca(2+)-ATPase pump or SERCA-2a whose function is to transport cytosolic Ca(2+) into the lumen of the SR during diastole and whose activity is regulated by reversible phosphorylation of the endogenously SR-bound phospholamban (PLB). The SR's terminal cisternea region contains ryanodine sensitive Ca(2+)-release channels (RR), the activity of which is regulated by direct and indirect reversible phosphorylation. These channels release the SR stored Ca(2+) during contraction. We have shown that in left ventricular (LV) myocardium from dogs with coronary microembolization-induced heart failure, ability of the SR to sequester and release Ca(2+) during the cardiac cycles is impaired. This abnormality was associated with reduced expression (protein and mRNA) levels of Ca(2+)-ATPase, PLB, and reduced PLB phosphorylation. Long-term therapy with the Acorn Cardiac Support Device (CSD) is associated with restoration of the ability of the SR to sequester Ca(2+). This improvement in SR function following chronic CSD therapy was due primarily to increased affinity of the SERCA-2a for calcium. The later was associated with (1) increased phosphorylation of PLB at serine 16 and threonine 17, (2) unchanged protein expression of PLB and (3) unchanged protein expression of SERCA-2a in LV myocardium of CSD-treated dogs compared to controls. This review summarizes our current understanding of the role of the CSD in modulating SR calcium cycling in an experimental canine model of chronic heart failure produced by multiple sequential intracoronary microembolizations. PMID- 16258726 TI - Integrin-dependent signal transduction regulating cyclin D1 expression and G1 phase cell cycle progression. AB - Integrins and growth factor receptors coordinately regulate proliferation in nontransformed cells. Coordinate signaling from these receptors controls the activation of the G1 phase cyclin-dependent kinases, largely by regulating levels of cyclin D1 and p27(kip1). Induction of cyclin D1 is one of the best understood examples of an integrin/growth factor receptor-regulated G1 phase target. This review focuses on the integrin-dependent signal transduction events that regulate the expression of cyclin D1 during G1 phase. PMID- 16258727 TI - Integrin-mediated activation of latent transforming growth factor beta. AB - Members of the integrin family recognize a variety of spatially-restricted extracellular ligands. Classically, ligation of integrins activates cytoplasmic signals in the integrin-expressing cell and contributes to cell adhesion, migration, proliferation and survival. At least two members of this family, alphavbeta6 and alphavbeta8 perform an additional function, activation of latent complexes of transforming growth factor beta. In effect, this process allows integrins on one cell to activate signals on adjacent (in the case of alphavbeta6) or nearby cells (in the case of alphavbeta8). Integrin-mediated TGFbeta activation has been shown to play important roles in modulating tissue fibrosis, acute lung injury and pulmonary emphysema. Given the important roles that TGFbeta plays in modulating epithelial cell growth, epithelial-to mesenchymal transformation and tumor invasion and metastasis, integrin-mediated TGFbeta activation is likely to play important roles in tumor growth and metastasis. PMID- 16258723 TI - Reversal of maladaptive gene program in left ventricular myocardium of dogs with heart failure following long-term therapy with the Acorn Cardiac Support Device. AB - Progressive left ventricular (LV) dilation is a characteristic feature of heart failure and is associated with poor long-term prognosis. One of the characteristic changes that occur in the failing heart is a change in gene expression wherein fetal genes that were turned off shortly after birth are re activated in heart failure and may play a key role in the progressive worsening of the heart failure state. This review discusses reversal of maladaptive gene expression in dogs with chronic heart failure treated long-term with the Acorn Cardiac Support Device (CSD); a passive mechanical device designed to prevent progressive LV enlargement and to restore normal LV chamber geometry. Studies in our laboratories have shown that, in addition to preventing LV dilation and improving LV ejection fraction, long-term therapy with the CSD reverses the maladaptive gene program observed in LV myocardium of dogs with heart failure. Therapy with the CSD was associated with up-regulated mRNA expression for alpha myosin heavy chain and down-regulated mRNA expression of A- and B- type natriuretic peptides, cytokines and favorably modulated cytoskeletal proteins. These findings provide an explanation for mechanisms that may be partly responsible for the improvement in LV systolic and diastolic function seen in dogs with heart failure after long-term CSD therapy. PMID- 16258728 TI - beta 1 integrin function in vivo: adhesion, migration and more. PMID- 16258729 TI - Mobilization and activation of a signaling competent alpha6beta4integrin underlies its contribution to carcinoma progression. AB - This review examines the hypothesis that the function of the alpha 6beta 4 integrin is altered substantially as normal epithelia undergo malignant transformation and progress to invasive carcinoma and that the functions of this integrin contribute to the behavior of aggressive carcinoma cells. Specifically, alpha 6beta 4 functions primarily as an adhesion receptor in normal epithelia, often as a component of hemidesmosomes and associated with intermediate filaments. Factors in the host-tumor microenvironment have the potential to mobilize alpha 6beta 4 from hemidesmosomes and promote its association with F actin in lamellae and filopodia, a process that is mediated by PKC-dependent phosphorylation of the beta 4 cytoplasmic domain. Importantly, this altered localization of alpha 6beta 4 appears to be coupled to an activation of its signaling potential, which may occur through its association with growth factor receptors or lipid rafts, possibilities that are not mutually exclusive. The primal signaling event triggered by alpha 6beta 4 appears to be activation of PI3 K and this activation has profound consequences on the migration, invasion and survival of carcinoma cells. Arguably, the ability of alpha 6beta 4 to stimulate the PI3-K-dependent translation of VEGF and possibly other growth factors may be the most significant contribution of this integrin to carcinoma because of the potential autocrine and paracrine effects of these factors. PMID- 16258730 TI - Clinging to life: cell to matrix adhesion and cell survival. AB - Cell to matrix adhesion regulates cellular homeostasis in multiple ways. Integrin attachment to the extracellular matrix mediates this regulation through direct and indirect connections to the actin cytoskeleton, growth factor receptors, and intracellular signal transduction cascades. Disruption of this connection to the extracellular matrix has deleterious effects on cell survival. It leads to a specific type of apoptosis known as anoikis in most non-transformed cell types. Anchorage independent growth is a critical step in the tumorigenic transformation of cells. Thus, breaching the anoikis barrier disrupts the cell's defenses against transformation. This review examines recent investigations into the molecular mechanisms of anoikis to illustrate current understanding of this important process. PMID- 16258732 TI - Milestones in the field of tumor microenvironment--contributions & perspectives of Prof. Isaac P. Witz. PMID- 16258731 TI - Review of Growth Inhibitory Peptide as a biotherapeutic agent for tumor growth, adhesion, and metastasis. AB - This review surveys the biological activities of an alpha-fetoprotein (AFP) derived peptide termed the Growth Inhibitory Peptide (GIP), which is a synthetic 34 amino acid segment produced from the full length 590 amino acid AFP molecule. The GIP has been shown to be growth-suppressive in both fetal and tumor cells but not in adult terminally-differentiated cells. The mechanism of action of this peptide has not been fully elucidated; however, GIP is highly interactive at the plasma membrane surface in cellular events such as endocytosis, cell contact inhibition and cytoskeleton-induced cell shape changes. The GIP was shown to be growth-suppressive in nine human tumor types and to suppress the spread of tumor infiltrates and metastases in human and mouse mammary cancers. The AFP-derived peptide and its subfragments were also shown to inhibit tumor cell adhesion to extracellular matrix (ECM) proteins and to block platelet aggregation; thus it was expected that the GIP would inhibit cell spreading/migration and metastatic infiltration into host tissues such as lung and pancreas. It was further found that the cyclic versus linear configuration of GIP determined its biological and anti-cancer efficacy. Genbank amino acid sequence identities with a variety of integrin alpha/beta chain proteins supported the GIP's linkage to inhibition of tumor cell adhesion and platelet aggregation. The combined properties of tumor growth suppression, prevention of tumor cell-to-ECM adhesion, and inhibition of platelet aggregation indicate that tumor-to-platelet interactions present promising targets for GIP as an anti-metastatic agent. Finally, based on cholinergic studies, it was proposed that GIP could influence the enzymatic activity of membrane acetylcholinesterases during tumor growth and metastasis. It was concluded that the GIP derived from full-length AFP represents a growth inhibitory motif possessing instrinsic properties that allow it to interfere in cell surface events such as adhesion, migration, metastasis, and aggregation of tumor cells. PMID- 16258733 TI - Group B streptococcus (Streptococcus agalactiae) meningitis in a diabetic adult. PMID- 16258734 TI - Alleviation of insulitis and moderation of diabetes in NOD mice following treatment with a synthetic Pseudomonas aeruginosa signal molecule, N-(3 oxododecanoyl)-L-homoserine lactone. AB - Quorum sensing signal molecules (QSSMs) from the bacterium Pseudomonas aeruginosa control bacterial population density and the expression of virulence determinants. Coincidentally, and possibly to allow this pathogen to gain a foothold in the human body, certain signal molecules also downregulate immunological responses in an apparently T-helper 1-selective manner, which would suggest their application as therapeutics to some autoimmune diseases. In the present paper, experiments are described that indicate that one particular signal molecule, a synthetic N-(3-oxododecanoyl)-L-homoserine lactone, can be used to alleviate insulitis and diabetes in non-obese diabetic (NOD) mice, suggesting that bacterial signal molecules may represent a novel source of immune modulatory compounds for the treatment of type 1 diabetes, which afflicts more than 2 million individuals in Europe and North America. PMID- 16258735 TI - Effect of haemodialysis on the oxidative stress and antioxidants in diabetes mellitus. AB - Oxidative stress has been defined as a loss of counterbalance between free radical or reactive oxygen species (ROS) production and antioxidant systems. It is involved in the pathogenesis of different chronic diseases. High levels of ROS production via different biochemical mechanisms accompany diseases like type 2 diabetes mellitus (DM) and end-stage renal disease (ESRD). Elevated oxidative status and reduced antioxidant defence systems in patients with DM and ESRD accelerate the prevalence of atherosclerosis and other chronic complications. Our aim was to reveal the effects of diabetes and haemodialysis (HD) separately and together on oxidative stress. In our study, we included 20 diabetic (DM) patients with no renal disease, 20 non-diabetic haemodialysis (HD), 20 diabetic haemodialysis (DHD) patients and 20 healthy volunteers. We have determined the levels of lipid peroxidation expressed as thiobarbituric acid-reactive substances (TBARS), oxidative protein damage as indicated by protein carbonyl (PCO) content and activities of antioxidant enzymes including superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSHPx) in all patient groups and healthy subjects. We found enhanced oxidative stress in all patient groups due to an increase in lipid peroxidation (TBARS) and increased oxidative protein damage in terms of PCO content and reduced activities of SOD, CAT and GSH-Px. Oxidative stress was more profound in diabetic patients undergoing haemodialysis. We conclude that both diabetes and dialysis increase oxidative stress and their combined effect on oxidative stress is the highest in magnitude as observed in diabetic patients undergoing haemodialysis. PMID- 16258736 TI - Factors associated with type 1 diabetes in Kuwaiti children. AB - Type 1 diabetes is a common chronic disease in childhood, and the outcome of environmental, genetic and immunologic interactions. The aim was to study the social and metabolic characteristics (lipids, lipoproteins, apolipoproteins, lipoprotein a (Lpa) and total sialic acid) and predisposing factors in 6-18-year old Kuwaiti children with type 1 diabetes. This pair-matched case-control study included 348 type 1 diabetic children (131 males, 217 females) matched by age and gender to 348 non-diabetic controls. Diabetic children were identified, according to the WHO and the American Diabetes Association criteria, at 182 randomly selected schools. Social and metabolic characteristics were adversely affected in diabetic children compared to their controls. The logistic regression analysis showed that the predisposing factors: family history of type 1 and type 2 diabetes and thyroid disease, were significant associated factors with type 1 diabetes after adjusting for demographic and social variables. The significant correlations of Lpa and total sialic acid with glycated haemoglobin, lipoproteins and apolipoproteins partially explain reporting them as possible markers for coronary heart disease. There are adverse metabolic changes in children with type 1 diabetes. As these changes are associated with early onset atherogenesis, metabolic markers need to be measured and possibly corrected at an early stage in children with diabetes. PMID- 16258737 TI - Differential p70S6k and 4E-BP1 regulation by insulin and amino acids in vascular endothelial and smooth muscle cells. AB - Differential stimulation of vascular endothelial and smooth muscle cells proliferation is responsible for atherosclerotic lesions. Amino acids and insulin modulate p70S6k and 4E-BP1 activity, regulating cell growth and proliferation. We hypothesised that nutritional (amino acids) and hormonal (insulin) signals differently modulate protein anabolism in human vascular endothelial (HUVEC) and smooth muscle (HVSMC) cells. We evaluated p70S6kinase and 4E-BP1 phosphorylation in the two cell types, grown in amino acid-free medium with or without insulin (INS, 100 nM) or/and amino acids mixture (AA, 3 mM) and with the selective addition or deprivation of branched chain amino acids (BCAA, 0.5 mM). INS stimulated p70S6k and 4E-BP1 phosphorylation transiently in HUVEC and persistently in HVSMC. AA and INS+AA stimulated p70S6k and 4E-BP1 phosphorylation persistently in HUVEC and HVSMC. AA, but not BCAA alone or BCAA-deprived AA, induced p70S6k phosphorylation in HUVEC. BCAA deprivation decreased the p70S6k phosphorylation induced by AA with or without insulin in HVSMC. These results show that anabolic stimuli modulate p70S6k and 4E-BP1 activity differently in the two vascular cell types, suggesting that insulin stimulates protein synthesis for a longer time in HUSMC than in HUVEC. We speculate that hyperinsulinaemia frequently associated with atherosclerosis could induce a selective HVSMC proliferation. PMID- 16258738 TI - Innervation and connexin isoform expression during diabetes-related bladder dysfunction: early structural vs. neuronal remodelling. AB - Urinary bladder dysfunction is a major complication in diabetes mellitus and its mechanism has been attributed to altered neurological function (autonomic and/or peripheral neuropathy). Previous studies have demonstrated impaired nerve deficiencies, including either loss of nerve function and/or anatomical loss of neuromuscular nerve terminals. While the phenomenon of diabetes-related neurological injury is well recognised, its pathogenesis is not well understood. Using a well established rat model of diabetes (streptozotocin model), we investigated the prevalence of sympathetic and parasympathetic nerves and relative prevalence of connexin isoforms (gap junction proteins) during diabetes related bladder dysfunction. Immunohistochemistry and digital image analysis was used to detect the prevalence of postsynaptic neuronal markers, NOS1 and connexin isoform expressions. Immunohistochemistry showed significant increases in tyrosine hydroxylase (marker of sympathetic innervation) and decreased vesicular acetylcholine transporter (marker of parasympathetic innervation), predominantly in the smooth muscle layer, 3 days after diabetes induction, when compared to age matched controls. Time-dependent and cell-specific decreases in the connexin 43 isoform, but transient increases in connexin 32 and 26, were also observed in diabetic rats vs. controls (p<0.05). These data suggest that selective and time dependent expression of gap junction proteins and altered prevalence of sympathetic/parasympathetic innervation are early events during diabetes-related bladder dysfunction and remodelling. PMID- 16258739 TI - Umpolung catalysts: comparative assessments on reactivities. AB - Umpolung catalysis is studied by a sequence of model reactions (CPCM in THF, B3LYP/6-31G*) with different aldehydes and catalysts. We involved addition of the catalyst to the aldehyde and 1,2-H-migration to form a carbanionic d1-species, which is the crucial intermediate according to the Lapworth- and Breslow mechanisms. Cyanide, N-methylthiazol-2-ylidene, and a glycol-based phosphite perform as umpolung catalysts, formaldehyde, acetaldehyde, benzaldehyde, and acrolein are substrates in this study. In these aldehyde substrates, alkyl substitution disfavors but pi-conjugation favors formation of the carbanionic d1 intermediate. The nucleophilic carbene, N-methylthiazol-2-ylidene, is the strongest umpolung catalyst, while the phosphite is about as active as cyanide. PMID- 16258740 TI - Ab initio computational insight into the ion-pair S(N)2 reaction of lithium isothiocyanate and methyl fluoride in the gas phase and in acetone solution. AB - The ion-pair S(N)2 reaction LiNCS + CH3F with two mechanisms, inversion and retention, was investigated at the MP2(full)/6-311+G**//HF/6-311+G** level in the gas phase and in acetone solution. All HF-optimized structures were confirmed by vibrational frequency analysis. Based on IRC analyses, eight possible reaction pathways in the title reaction are proposed. The inversion mechanism through a six-membered-ring transition-state structure is the most favorable. Methyl thiocyanate should form preferentially in the gas phase and the more stable methyl isothiocyanate will be the main product in CH3COCH3. The retardation of the reaction in CH3COCH3 solution was attributed to the differences in the solvation free energies in the separated reactants and transition structures. All of the theoretical results are consistent with the experiment. PMID- 16258741 TI - Classifying wine according to geographical origin via quadrupole-based ICP-mass spectrometry measurements of boron isotope ratios. AB - The potential of quadrupole-based ICP-MS as a tool for B-isotopic analysis of wines and its usefulness in provenance determinations were assessed. A precision of 0.1-0.25% RSD (corresponding to a relative standard deviation of the mean of three replicate measurements of 0.06-0.12%) was sufficient to establish small differences in the B isotope ratios in wines from different geographical origins. Each sample measurement was bracketed by measurements of a standard and mass bias drift correction made by interpolation. Sample preparation was kept to a minimum to avoid possible fractionation. Dilution of the wine samples by a factor of 100 with 0.65% HNO3 was found to reduce matrix-induced mass discrimination substantially. Wines from three wine-producing regions, Stellenbosch, Robertson, and Swartland, in the Western Cape Province of South Africa, and wines from specific regions in France (Bergerac) and Italy (Valpolicella) were analyzed by ICP-QMS for their B-isotopic compositions. It was concluded that the 11B/10B ratios can be used to characterize wines from different geographical origins. Average 11B/10B ratios in red wines from South Africa (Stellenbosch), France (Bergerac), and Italy (Valpolicella) were found to differ by between 0.5 and 1.5%. PMID- 16258742 TI - New type of dry substances content meter using microwaves for application in biogas plants. AB - Dry substances (DS) are an important index for monitoring and controlling anaerobic co-digestion in biogas plants. We have developed and tested an online meter that measures suspended solids by means of the reflection coefficient of an exiting microwave signal, which is dependent on the dielectric properties of the suspensions. Intelligent models based on partial least squares regression (PLSR) and artificial neural network (ANN) for calibration allow exact and reproducible measurements under different circumstances. This measuring method is appropriate for contactless and online measurements of dry substance contents in biogas plants in a large range from 2-14%. PMID- 16258744 TI - Neutropenic enterocolitis (typhlitis) associated with infectious mononucleosis. AB - Neutropenic enterocolitis (typhlitis) is an unusual acute complication of neutropenia, most often associated with leukaemia and lymphoma and characterized by segmental caecal and ascending colonic ulceration that may progress to necrosis, perforation, and septicaemia. We present a unique case of an 8-year-old girl with recently diagnosed infectious mononucleosis having findings consistent with typhlitis on abdominal CT. PMID- 16258743 TI - ATP-loaded liposomes effectively protect the myocardium in rabbits with an acute experimental myocardial infarction. AB - PURPOSE: We assessed whether the infusion of ATP-loaded liposomes (ATP-L) can limit the fraction of the irreversibly damaged myocardium in rabbits with an experimental myocardial infarction. METHODS: ATP-L, empty liposomes (EL), or Krebs-Henseleit (KH) buffer were administered by intracoronary infusion, followed by 30 min of occlusion and 3 h of reperfusion. Unisperse Blue dye was used to demarcate the net size of the occlusion-induced ischemic zone (area at risk) and nitroblue tetrazolium staining was used to detect the final fraction of the irreversibly damaged myocardium within the total area at risk. RESULTS: The total size of the area at risk in all experimental animals was approx. 20% wt. of the left ventricle. The final irreversible damage in ATP-L-treated animals was only ca. 30% of the total area at risk as compared with ca. 60% in the group treated with EL (p < 0.009) and ca. 70% in the KH buffer-treated group (p < 0.003). CONCLUSIONS: ATP-L effectively protected the ischemic heart muscle in rabbits with an experimental myocardial infarction as evidenced by a significantly decreased fraction of the irreversibly damaged heart within the total area at risk. ATP-L may provide an effective exogenous source of the ATP in vivo to protect ischemically damaged cells. PMID- 16258745 TI - Effect of short course high dose methylprednisolone on endothelin-1 and nitric oxide in children with acute lymphoblastic leukemia: a preliminary study. AB - We have investigated serum endothelin-1 (ET) and nitric oxide (NO) levels before and after a short course of high dose methylprednisolone (HDMP) in children with acute lymphoblastic leukemia (ALL) as an indicator of vasoconstrictor and vasodilator properties of endothelium. Nineteen children with ALL (aged 13-180 months; 5 girls and 14 boys) and 25 healthy children were included in the present study. The children with ALL were given HDMP (20 mg kg 1 day 1) alone for the first 5 days. Serum ET and NO levels were analysed before and after a short course of high dose methylprednisolone. Before treatment, serum ET levels (median 7.6 pg ml 1) of the patients were lower than the healthy controls (13.0 pg ml 1) (p < 0.05), and it rose to similar levels (13.0 pg ml 1) following therapy as in the controls. Nitric oxide levels (7.0 micromol) of the patients were insignificantly higher than the healthy controls (3.9 micromol) and did not differ after treatment (7.0 micromol) (p > 0.05). In conclusion, the elevation of ET to normal level following treatment suggests that a short course of high dose of methylprednisolone improve the endothelial dysfunction caused by acute leukemia. PMID- 16258746 TI - Kinematics and motor activity during tethered walking and turning in the cockroach, Blaberus discoidalis. AB - When insects turn from walking straight, their legs have to follow different motor patterns. In order to examine such pattern change precisely, we stimulated single antenna of an insect, thereby initiating its turning behavior, tethered over a lightly oiled glass plate. The resulting behavior included asymmetrical movements of prothoracic and mesothoracic legs. The mesothoracic leg on the inside of the turn (in the apparent direction of turning) extended the coxa trochanter and femur-tibia joints during swing rather than during stance as in walking, while the outside mesothoracic leg kept a slow walking pattern. Electromyograms in mesothoracic legs revealed consistent changes in the motor neuron activity controlling extension of the coxa-trochanter and femur-tibia joints. In tethered walking, depressor trochanter activity consistently preceded slow extensor tibia activity. This pattern was reversed in the inside mesothoracic leg during turning. Also for turning, extensor and depressor motor neurons of the inside legs were activated in swing phase instead of stance. Turning was also examined in free ranging animals. Although more variable, some trials resembled the pattern generated by tethered animals. The distinct inter joint and inter-leg coordination between tethered turning and walking, therefore, provides a good model to further study the neural control of changing locomotion patterns. PMID- 16258747 TI - Polar-localised poplar K+ channel capable of controlling electrical properties of wood-forming cells. AB - In previous studies, we have shown that annual expression profiles of cambial and wood tissue with respect to the Shaker K+ channel PTORK correlate with cambial activity. To follow PTORK-gene activity on the cellular level, we isolated the respective promoter regions and generated transgenic Arabidopsis plants expressing the GUS gene under the control of the PTORK promoter. Cross-sections of petioles showed PTORK-driven signals predominantly in the xylem parenchyma surrounding the vessels and in the phloem. Antibodies raised against a unique N terminal region of PTORK in histo-immunochemical analyses recognised this K+ release channel in growth-active poplar plants only. PTORK labelling was found in differentiating xylem cells (young fibres) and mature xylem (vessel-associated cells of the ray parenchyma). Patch-clamp measurements on fibre cell protoplasts, derived from young poplar twigs, identified outward-rectifying K+ channels as the major K+ conductance of this cell type, which resembled the biophysical properties of PTORK when expressed in Xenopus oocytes. PMID- 16258748 TI - How much are health-care systems prepared to pay to produce a QALY? PMID- 16258749 TI - Definition of the "health benefit basket" in poland. AB - The subject of "health benefit basket" has been hotly debated for years among the Polish public, but until recently the debate has tended to be largely theoretical and abstract and therefore has lacked an effect on public policy. The situation changed in 2004, for two reasons: first the verdict of the Constitutional Tribunal invalidating the existing health insurance law and, second, Poland's accession to the European Union. The first problem was solved in part by defining a list of specific exclusions in the law and a promise to establish an institution for health technology assessment. The second issue remains open, although to some extend it is being dealt with legally by regulations issued from the Ministry of Health on acceptable waiting times for health services. PMID- 16258750 TI - The "Essential Levels of Care" in Italy: when being explicit serves the devolution of powers. AB - The definition of an explicit health benefit package in Italy has gained importance because of devolution of powers from the national level to the regions. The set of services to be guaranteed by the public sector are defined at national level, while regions are accountable for their provision. This contribution discusses the entitlements and the decision criteria adopted by Italian policy-making bodies. Entitlements to services are clearly defined for few sectors (mainly outpatient specialist care); for hospital care the benefit catalogue is vague. The definition of the health benefit package in Italy is an essential element of the relationship between the central government and the regions. It is argued that adequate monitoring systems and accountability procedures are still needed to make the essential levels of care an effective pivotal element of the Italian National Health Service. PMID- 16258752 TI - Correlation between burnout syndrome and psychological and psychosomatic symptoms among teachers. AB - OBJECTIVES: Psychosomatic disorders and symptoms that correlate with the so called burnout syndrome turned out to be the main cause of increasing rates of premature retirement of school teachers. The aim of this study was to evaluate the relation between occupational burden and psychological strain of teachers who are still in work. METHODS: A sample of 408 teachers at ten grammar schools (am: High school; German: Gymnasium) in south-western Germany was evaluated. To determine the styles of coping with occupational burden we used the measure of coping capacity questionnaire (MECCA). To analyse the psychopathological and psychosomatic symptom load we applied SCL 90 R questionnaire. RESULTS: According to the MECCA questionnaire, 32.5% of the sample suffered from burnout (type B), 17.7% suffered severe strain (type A), 35.9% showed an unambitious (type S) and 13.8% showed a healthy-ambitious coping style (type G). Burnout was significantly higher among women, divorced teachers and teachers working part-time. As part of the MECCA, teachers were asked to rate what they regarded as the strongest factor resulting in occupational burden. Teachers indicated that, besides high numbers of pupils in one class, they regarded destructive and aggressive behaviour of pupils as the primary stress factor. According to the SCL 90 R, 20% of the sample showed a severe degree (defined as >70 points in the SCL90R GSI) of psychological and psychosomatic symptoms. MECCA type B (burnout) correlated significantly with high psychological and psychosomatic symptom load according to the SCL90R. CONCLUSIONS: In school teachers, burnout syndrome, a construct that derived from occupational psychology and occupational medicine, is significantly correlated with psychological and psychosomatic symptoms. Teachers rate destructive and aggressive behaviour of pupils as the primary stress factor. PMID- 16258751 TI - Effects of fluoxetine and buspirone on the panicolytic-like response induced by the activation of 5-HT1A and 5-HT2A receptors in the rat dorsal periaqueductal gray. AB - RATIONALE: Administration of 5-hydroxytryptamine (5-HT)1A and 5-HT2A receptor agonists into the dorsal periaqueductal gray (DPAG) inhibits escape, a defensive behavior associated with panic attacks. Long-term treatment with the antipanic compound imipramine enhances the DPAG 5-HT1A- and 5-HT2A-receptor-mediated inhibition of escape, implicating these receptors in the mode of action of panicolytic drugs. OBJECTIVES: In the present study, we investigated whether the inhibitory effect on escape elicited by the intra-DPAG injection of 5-HT1A and 5 HT2A receptor agonists is also enhanced after treatment with fluoxetine, another widely used antipanic drug. The effects of fluoxetine were compared to those of buspirone, an anxiolytic drug without major effect on panic disorder. METHODS: Male Wistar rats, subchronically (3-6 days) or chronically (21-24 days) treated with fluoxetine (10 mg/kg i.p.) or chronically treated with buspirone (0.3 mg/kg i.p.), were intra-DPAG injected with 5-HT (20 nmol), the 5-HT1A receptor agonist (+/-)-8-hydroxy-2-(di-n-propylamino)tetralin hydrobromide (8-OH-DPAT; 8 nmol) or the preferential 5-HT2A receptor agonist (+/-)-1-(2,5-dimethoxy-4-iodophenyl) piperazine dihydrochloride (DOI; 16 nmol). The intensity of electrical current that applied to the DPAG-evoked escape behavior was measured before and after the microinjection of these agonists. RESULTS: The electrical current necessary to produce escape was increased after the microinjection of the three 5-HT receptor agonists in all groups of animals tested. However, this panicolytic-like effect was significantly higher in animals receiving long-term treatment with fluoxetine. CONCLUSIONS: The results suggest that facilitation of the 5-HT1A- and 5-HT2A-receptor-mediated inhibition of DPAG neuronal activity is implicated in the beneficial effect of antidepressants in panic disorder. PMID- 16258753 TI - SMOOTH: a statistical method for successful removal of genotyping errors from high-density genetic linkage data. AB - High-density genetic linkage maps can be used for purposes such as fine-scale targeted gene cloning and anchoring of physical maps. However, their construction is significantly complicated by even relatively small amounts of scoring errors. Currently available software is not able to solve the ordering ambiguities in marker clusters, which inhibits the application of high-density maps. A statistical method named SMOOTH was developed to remove genotyping errors from genetic linkage data during the mapping process. The program SMOOTH calculates the difference between the observed and predicted values of data points based on data points of neighbouring loci in a given marker order. Highly improbable data points are removed by the program in an iterative process with a mapping algorithm that recalculates the map after cleaning. SMOOTH has been tested with simulated data and experimental mapping data from potato. The simulations prove that this method is able to detect a high amount of scoring errors and demonstrates that the program enables mapping software to successfully construct a very accurate high-density map. In potato the application of the program resulted in a reliable placement of nearly 1,000 markers in one linkage group. PMID- 16258754 TI - [Clinical trials for bladder cancer in Germany]. AB - The urothelial cancer section of the Association of Urogenital Oncology (AUO) of the German Cancer Society has successfully performed and published clinical trials on treatment of urothelial neoplasia. During the last decade, around 770 patients have been recruited for several phase I, II and III trials. The majority of trials had been conceived for advanced inoperable stages, but the AUO has also been particularly successful in recruiting large trials on adjuvant chemotherapy after radical cystectomy for locally advanced bladder cancer. Future goals address extending the number of active centers in Germany as well as seeking international collaborations in order to enable timely recruitment of trials that demand an extensive sample size. PMID- 16258755 TI - [Functional magnetic resonance imaging of the gastrointestinal tract--clinical application possibilities?]. AB - Magnetic resonance imaging (MRI) is a versatile medical imaging tool for which several new applications have been developed. Beside its broad clinical use for the detection of anatomical structures and pathologies MRI has been successfully applied for the non-invasive imaging of human organ functions, including the brain and the cardiovascular system. The use of MRI for the assessment and analysis of gastrointestinal (GI) function is a new approach that is currently performed in only a few research sites. Several characteristics make MRI an ideal technique for the direct assessment of GI physiology: MRI acquires high resolution images with excellent soft tissue contrast, it does not expose subjects to ionizing radiation, is non-invasive, and the acquisition and analysis of the images can be independently verified. In this article we summarize recent developments of MRI techniques in GI research. We will also discuss the advantages and limitations of MRI for this purpose in relation to established medical imaging tools and investigations. PMID- 16258757 TI - Response to Esteller et al. PMID- 16258756 TI - Cyclin D1 gene (CCND1) polymorphism and the risk of squamous cell carcinoma of the larynx. AB - Cyclin D1 is one of the key proteins involved in cell cycle control, and it is believed that its overexpression may be connected with tumorigenesis. A reason for cyclin D1 deregulation may be connected to a common G870A polymorphism at codon 242 in exon 4 of the CCND1 gene. This single nucleotide substitution, localized in the conserved splice donor site between exon 4 and the intron 4 boundary, might modulate the frequency of alternative splicing. It has been postulated that the A allele results in a higher level of mRNA (transcript b) encoding a protein with an altered C-terminal domain. The influence of CCND1 G- >A polymorphism for the risk of cancer and the prognosis of patients with different types of solid tumors has already been suggested. This study was conducted to investigate the association between the cyclin D1 gene polymorphism and laryngeal cancer risk, as well as the clinical outcome. We also examined the relationship between genotype/allele distributions and the cyclin D1 expression profile. The genotyping study was done using the PCR-RFLP method in 63 patients with larynx cancer and 102 healthy controls. The heterozygotic genotype GA as well as a combination of GA and AA genotypes were associated with an increased risk of larynx cancer compared to the GG genotype (OR =3.02; P =0.004 and OR =2.52; P =0.013, respectively). The A allele frequency was higher in cancer cases (0.484) than in controls (0.416) that were connected with a slightly increased risk of cancer development (OR =1.34); however, the difference was not significant. The AA genotype was associated with an early cancer onset compared to the GG genotype (median age: 51.5 and 63.0 years, respectively). We also demonstrated that the AA genotype was associated with the occurrence of lymph node metastases (OR =3.26) and a higher level of cyclin D1 overexpression. These results suggest that the CCND1 A allele may be a genetic factor that modulates the risk of larynx cancer development, and it may also have an effect on tumor biology and disease prognosis. PMID- 16258758 TI - Prognostic factors in laryngotracheal injury following intubation and/or tracheotomy in ICU patients. AB - The aim of this study was to determine the incidence of laryngotracheal injuries following intubation and/or tracheotomy in intensive care unit (ICU) patients and to analyze their prognostic factors. This prospective study includes the clinical data and endoscopic exploration of 654 ICU patients who underwent oro-tracheal intubation between September 1992 and February 1999. The prognostic factors for upper airway injuries were analyzed using a multivariate statistical study. Endoscopic exploration of the upper airway 6 to 12 months after extubation revealed laryngotracheal injuries in 30 of the 280 patients examined (11%). The most important factors influencing the development of laryngotracheal lesions were the duration of the oro-tracheal intubation and the length of time in the ICU. Patients at high risk of developing injuries were those with pathological background, a non-neurological or non-surgical (medical) admission or upper airway injuries at an early stage. The length of oro-tracheal intubation is the most important factor in the development of laryngotracheal injuries. Consequently, it is essential to establish a time limit to perform tracheotomy in ICU patients. Such timing should be adapted to each patient and pathology. PMID- 16258759 TI - [Efficacy and tolerability of topical foscarnet sodium in treatment of herpes labialis. Results of post-marketing surveillance]. AB - BACKGROUND AND OBJECTIVE: Aim of this post-marketing surveillance study was to generate further data on efficacy and tolerability of foscarnet sodium cream 2% in patients with recurrent herpes labialis in daily routine practice. DESIGN AND METHODS: 90 outpatients with recurrent herpes labialis were instructed to apply the antiviral cream to the lesions according to package insert for up to 10 days. Symptoms were recorded before beginning of treatment, after three days, and at the end of treatment. RESULTS: Paresthesia and swelling as well as the number of vesicles were reduced after three days of observation, and were improved by about 90% at the end of treatment. Erosions and crust formation also improved markedly. Efficacy as well as tolerability were evaluated as positive by the majority of physicians and patients. No adverse reactions were reported. CONCLUSIONS: Foscarnet sodium 2% cream is an effective and safe medication for the treatment of recurrent labial herpes in daily routine practice of dermatologists. PMID- 16258760 TI - Immunoglobulin KM allotypes are associated with the prevalence of autoantibodies to GD1a ganglioside, but not with susceptibility to the disease, in Japanese patients with Guillain-Barre syndrome. AB - Guillain-Barre syndrome (GBS), an autoimmune disease of the peripheral nervous system, is associated with antecedent Campylobacter jejuni infection. GM and KM allotypes--genetic markers of immunoglobulin gamma and kappa chains, respectively -are implicated in the etiopathogenesis of several autoimmune diseases. To determine if GM/KM phenotypes are associated with GBS and influence antibody responses to C. jejuni and to GM1 and GD1a gangliosides, 72 Japanese GBS patients and 73 controls were allotyped for several GM and KM markers. Sera from patients were characterized for antibodies to C. jejuni, GM1, and GD1a. The distribution of KM phenotypes was significantly different in patients with anti-GD1a ganglioside antibodies from those who lacked these antibodies (P=0.029). No other significant associations were found. These results suggest that KM allotypes are not risk factors for developing GBS, but contribute significantly to the generation of autoimmune responses to GD1a ganglioside in patients with this disease. PMID- 16258761 TI - Clinical molecular imaging and therapy-moving ahead together. PMID- 16258762 TI - NanoPET imaging of [(18)F]fluoromisonidazole uptake in experimental mouse tumours. AB - PURPOSE: The purpose of this study was to assess the potential and utility of ultra-high-resolution hypoxia imaging in various murine tumour models using the established hypoxia PET tracer [(18)F]fluoromisonidazole ([(18)F]FMISO). METHODS: [(18)F]FMISO PET imaging was performed with the dedicated small-animal PET scanner NanoPET (Oxford Positron Systems) and ten different human tumour xenografts in nude mice as well as B16 melanoma tumours in syngeneic Balb/c mice. For comparison, [(18)F]fluorodeoxyglucose ([(18)F]FDG) PET scans were also performed in the mice bearing human tumour xenografts. RESULTS: In 10 out of 11 experimental tumour models, [(18)F]FMISO PET imaging allowed clear-cut visualisation of the tumours. Inter- and intratumoural heterogeneity of tracer uptake was evident. In addition to average TMRR (tumour-to-muscle retention ratio including all voxels in a volume of interest (VOI)), the parameters TMRR(75%) and TMRR(5) (tumour-to-muscle retention ratio including voxels of 75% or more of the maximum radioactivity in a VOI and the five hottest pixels, respectively) also served as measures for quantifying the heterogeneous [(18)F]FMISO uptake in the tumours. The variability observed in [(18)F]FMISO uptake was related neither to tumour size nor to the injected mass of the radiotracer. The pattern of normoxic and hypoxic regions within the human tumour xenografts, however, correlated with glucose metabolism as revealed by comparison of [(18)F]FDG and [(18)F]FMISO images. CONCLUSION: This study demonstrates the feasibility and utility of [(18)F]FMISO for imaging murine tumour models using NanoPET. PMID- 16258763 TI - Site-specific conjugation of HIV-1 tat peptides to IgG: a potential route to construct radioimmunoconjugates for targeting intracellular and nuclear epitopes in cancer. AB - PURPOSE: Our objective was to study the cellular and nuclear uptake of (123)I mouse IgG ((123)I-mIgG) linked to peptides [GRKKRRQRRRPPQGYGC] harbouring the membrane-translocating and nuclear import sequences of HIV-1 tat protein. METHODS: Carbohydrates on mIgG were oxidized by NaIO(4), then reacted with a 40 fold excess of peptides. Displacement of binding of anti-mouse IgG (Fab specific; alpha-mFab) to (123)I-mIgG by tat-mIgG or mIgG was compared. Internalization and nuclear translocation of (123)I-tat-mIgG in MDA-MB-468, MDA-MB-231 or MCF-7 breast cancer cells were measured. The immunoreactivity of imported tat-mIgG was evaluated by measuring binding of (123)I-alpha-mFab to cell lysate and by displacement of binding of (123)I-mIgG to alpha-mFab by cell lysate. Biodistribution and nuclear uptake of (123)I-tat-mIgG, (123)I-mIgG and (123)I-tat were compared in mice bearing s.c. MDA-MB-468 tumours. RESULTS: There was a 15 fold decrease in affinity of alpha-mFab for tat-mIgG compared with mIgG. Internalized radioactivity imported into the nucleus for (123)I-tat-mIgG in MDA MB-468, MDA-MB-231 and MCF-7 cells was 61.5+/-0.6%, 60.3+/-3.6% and 64.7+/-1.0%, respectively. The binding of (123)I-alpha-mFab to lysate from MDA-MB-468 cells importing tat-mIgG was 17-fold higher than that for cells not exposed to tat mIgG. Imported tat-mIgG competed with tat-mIgG for displacement of binding of (123)I-mIgG to alpha-mFab. Conjugation of mIgG to tat peptides did not change tissue distribution. Nuclear localization for (123)I-tat-mIgG in MDA-MB-468 tumours was 28.1+/-5.6%, and for liver, spleen and kidneys it was 41.7+/-2.7%, 13.8+/-0.8% and 36.9+/-3.3%, respectively. CONCLUSION: (123)I-tat-mIgG radioimunoconjugates suggest a route to the design of radiopharmaceuticals exploiting intracellular and nuclear epitopes. PMID- 16258765 TI - Initial experience with FDG-PET/CT in the evaluation of breast cancer. AB - PURPOSE: We retrospectively reviewed FDG-PET/CT images in patients with breast cancer to determine whether PET/CT improved the level of diagnostic confidence as compared with PET and to compare PET/CT and CT findings at the location of suspected malignancies. METHODS: The study included 75 patients with known breast cancer. The initial PET/CT study for each patient was retrospectively reviewed to determine whether improved diagnostic confidence (IDC) regarding lesion localization and characterization was observed with PET/CT as compared with PET alone. PET/CT and CT findings were compared regarding lesion characterization and staging in 69 of the 75 patients, and in the case of discordant findings, comparison with histological or informative follow-up results was also performed. RESULTS: Fifty of the 75 patients exhibited increased FDG uptake in a total of 95 regions. In the comparison of PET/CT and PET, PET/CT resulted in IDC in 30 (60%) of these 50 patients and in 52 (55%) of the 95 regions. In the comparison between PET/CT and CT in 69 patients, PET/CT demonstrated a significantly better accuracy than CT (P<0.05). PET/CT showed definitely positive findings in 60 regions with malignancies, among which CT exhibited positive findings in 43 (72%). PET/CT and CT accurately staged 59 (86%) and 53 (77%) of the 69 patients, respectively. CONCLUSIONS: PET/CT added incremental diagnostic confidence to PET in more than 50% of patients and regions with increased FDG uptake. PET/CT accurately detected more regions with malignancies than did CT. This initial evaluation suggests that PET/CT is preferable to PET or CT in the diagnosis of breast cancer. PMID- 16258764 TI - A comparison of PET imaging characteristics of various copper radioisotopes. AB - PURPOSE: PET radiotracers which incorporate longer-lived radionuclides enable biological processes to be studied over many hours, at centres remote from a cyclotron. This paper examines the radioisotope characteristics, imaging performance, radiation dosimetry and production modes of the four copper radioisotopes, ( 60)Cu,( 61)Cu,( 62)Cu and( 64)Cu, to assess their merits for different PET imaging applications. METHODS: Spatial resolution, sensitivity, scatter fraction and noise-equivalent count rate (NEC) are predicted for( 60)Cu,( 61)Cu,( 62)Cu and( 64)Cu using a model incorporating radionuclide decay properties and scanner parameters for the GE Advance scanner. Dosimetry for( 60)Cu,( 61)Cu and( 64)Cu is performed using the MIRD model and published biodistribution data for copper(II) pyruvaldehyde bis(N(4) methyl)thiosemicarbazone (Cu-PTSM). RESULTS: (60)Cu and( 62)Cu are characterised by shorter half-lives and higher sensitivity and NEC, making them more suitable for studying the faster kinetics of small molecules, such as Cu-PTSM.( 61)Cu and( 64)Cu have longer half-lives, enabling studies of the slower kinetics of cells and peptides and prolonged imaging to compensate for lower sensitivity, together with better spatial resolution, which partially compensates for loss of image contrast.( 61)Cu-PTSM and( 64)Cu-PTSM are associated with radiation doses similar to [(18)F]-fluorodeoxyglucose, whilst the doses for( 60)Cu-PTSM and( 62)Cu-PTSM are lower and more comparable with H(2) (15)O. CONCLUSION: The physical and radiochemical characteristics of the four copper isotopes make each more suited to some imaging tasks than others. The results presented here assist in selecting the preferred radioisotope for a given imaging application, and illustrate a strategy which can be extended to the majority of novel PET tracers. PMID- 16258769 TI - Conflict in biomedical research. PMID- 16258770 TI - Foreword--focused issue on cardiac repair by stem cells. PMID- 16258766 TI - Regulation of cardiac inwardly rectifying potassium current IK1 and Kir2.x channels by endothelin-1. AB - To elucidate the ionic mechanism of endothelin-1 (ET-1)-induced focal ventricular tachyarrhythmias, the regulation of I(K1) and its main molecular correlates, Kir2.1, Kir2.2 and Kir2.3 channels, by ET-1 was investigated. Native I(K1) in human atrial cardiomyocytes was studied with whole-cell patch clamp. Human endothelin receptors were coexpressed with human Kir2.1, Kir2.2 and Kir2.3 channels in Xenopus oocytes. Currents were measured with a two-microelectrode voltage clamp. In human cardiomyocytes, ET-1 induced a marked inhibition of I(K1) that could be suppressed by the protein kinase C (PKC) inhibitor staurosporine. To investigate the molecular mechanisms underlying this regulation, we studied the coupling of ET(A) receptors to homomeric and heteromeric Kir2.1, Kir2.2 and Kir2.3 channels in the Xenopus oocyte expression system. ET(A) receptors coupled functionally to Kir2.2 and Kir2.3 channels but not to Kir2.1 channels. In Kir2.2 channels lacking functional PKC phosphorylation sites, the inhibitory effect was abolished. The inhibition of Kir2.3 currents could be suppressed by the PKC inhibitors staurosporine and chelerythrine. The coupling of ET(A) receptors to heteromeric Kir2.1/Kir2.2 and Kir2.2/Kir2.3 channels resulted in a strong inhibition of currents comparable with the effect observed in Kir2.2 homomers. Surprisingly, in heteromeric Kir2.1/Kir2.3 channels, no effect was observed. ET-1 inhibits human cardiac I(K1) current via a PKC-mediated phosphorylation of Kir2.2 channel subunits and additional regulatory effects on Kir2.3 channels. This mechanism may contribute to the intrinsic arrhythmogenic potential of ET-1. PMID- 16258775 TI - Near syncope while chewing food. PMID- 16258774 TI - Hypertriglyceridemia-why, when and how should it be treated? AB - The relationship between serum triglyceride levels and cardiovascular disease has remained enigmatic despite four decades of research. The majority of the available evidence tends to support the role of hypertriglyceridemia as an independent risk factor for cardiovascular disease. However, there are no guidelines recommending a target triglyceride level for prevention of cardiovascular disease. The focus of lipid lowering therapy still remains the reduction of global cardiovascular risk by optimizing LDL cholesterol levels. Therapeutic options for triglyceride-lowering include lifestyle modification and pharmacological agents, such as fibrates, omega 3 fatty acids and niacin. Post hoc analyses of the Helsinki Heart Study, Veterans Affairs High-Density Lipoprotein Cholesterol Intervention Trial and Bezafibrate Infarction Prevention Study suggest a beneficial effect of the treatment of hypertriglyceridemia with fibrates, mainly in obese subjects with insulin resistance. However, in order to establish the actual clinical relevance of lowering triglyceride levels, prospective trials need to be conducted with the specific purpose of studying the effects of triglyceride reduction on clinical end points, i. e. coronary events and stroke. PMID- 16258776 TI - Treatment of in-stent restenosis with sirolimus-eluting-stents -- a six month clinical and angiographic follow-up. AB - Treatment of in-stent restenosis (ISR) remains a therapeutic challenge since many pharmacological and mechanical approaches have shown disappointing results except for brachytherapy. Drug-eluting stents (DES) have been reported to effectively reduce ISR in de novo lesions. We studied 55 consecutive patients with ISR in native coronary arteries and 7 with ISR in saphenous vein grafts (SVG) with elective indication for percutaneous coronary intervention (PCI), who underwent successful implantation with DES. No in-hospital postprocedural major adverse cardiac events were observed. All but one patient (n=61) underwent an angiographic follow-up at 183+/-30 days. Grade of stenosis was assessed by quantitative coronary angiography (QCA) at index procedure and at control angiography. Restenosis (>50%) occurred in 5 patients (8.2%). Target vessel revascularization was performed in an additional 4 patients. Minimal intimal hyperplasia was observed in all segments covered by DES (late loss 0.08+/-0.37 mm, loss index 0.11+/-0.47). One patient suffered from subacute stent thrombosis due to discontinuation of clopidogrel medication. At six month follow-up two patients had died. Death was not related to a restenosis in the treated segment. Conclusion Our experiences with DES treatment of ISR lesions show good angiographic and clinical results at index procedure and at the 6 month follow-up with low sub acute thrombosis rate as compared with existing treatment modalities. Restenosis rate seems to be at least as low as reported for brachytherapy. PMID- 16258777 TI - Risk stratification models fail to predict hospital costs of cardiac surgery patients. AB - BACKGROUND: The aim of this prospective study was to determine if commonly used risk stratification models can predict total hospital costs in cardiac surgical patients. METHODS: Between October 1st and December 31st 2003, all consecutive adult patients undergoing cardiac surgery on CPB at our institution were classified using seven risk stratification scoring systems: EuroSCORE, Cleveland, Parsonnet, Ontario, French, Pons, and CABDEAL. Total hospital costs for each patient were calculated on a daily basis including preoperative diagnostic tests, operating room costs, disposable materials, drugs, blood components, costs for personnel, and hospital fixed-costs. Linear regression analysis was used to determine the correlation between costs and the seven risk stratifications models as well as length of stay (LOS) on ICU. The Spearman correlation coefficient was calculated from the regression line, and an analysis of residuals was performed to determine the quality of the regression. RESULTS: A total of 252 patients were operated for CABG (n=175), valve (n=39), CABG plus valve (n=21), thoracic aorta (n=13) and miscellaneous (2 myxoma, 1 ASD, 1 pulmonary embolism). Mean age of the patients was 66.0+/-11.4 years, 29.4% were female. LOS on ICU was 3.3+/-6.3 days and the 30-day mortality rate was 6.7%. Spearman correlation between the seven risk stratification models and hospital costs was below r=0.32 (p=0.0001), but was r=0.94 (p=0.0001) between ICU LOS and costs. CONCLUSIONS: Total hospital costs can be identified by length of ICU stay. None of the common risk stratification models accurately predicted total hospital costs in cardiac surgical patients. PMID- 16258778 TI - Female patient with proximal myotonic myopathy and ventricular tachycardia. AB - A 68-year-old woman with known proximal myotonic myopathy was transferred to our hospital for further diagnostic and therapeutic evaluation after successful termination of an episode of sustained ventricular tachycardia. In 2001, the myopathy was diagnosed after symptomatic weakness of the hip flexors. A cardiomyopathy with slight reduction of systolic left ventricular function was found in 2002. Coronary angiography excluded relevant coronary artery disease. The electrophysiologic examination could provoke atrial flutter, but neither a ventricular tachycardia nor a delay in the AV conduction was found. ECG and Holter ECG did not reveal any abnormalities. A reduction of the left ventricular systolic function (EF 45%) with normal size of cardiac chambers was demonstrated by echocardiography. It is known that in the patient group with myotonic dystrophies cardiac involvement manifests itself as cardiomyopathy, conduction disturbance or arrhythmia. However, only a small percentage of all patients with myotonic myopathy actually suffer from cardiac involvement. Among the different types of cardiac involvement, conduction disturbances requiring pacemaker implantation are most frequent. Only some patients develop ventricular tachycardias, and even cases of sudden cardiac death have been described. As a result of the case reports in the literature and the findings in our patient an ICD system was implanted on March 4, 2004. PMID- 16258780 TI - Impedance cardiography and beat-to-beat blood pressure monitoring in diagnosis of syncope in long-QT syndrome. AB - Syncope represents a diagnostic challenge in patients affected by long-QT syndrome (LQTS). It is crucial for the therapeutic decision to distinguish between potentially life-threatening ventricular tachycardias (Torsadede-pointes) and-especially during adolescence-neurocardiogenic syncopes. This case report presents a patient with LQTS2 (mutation in the HERG gene) on medication with beta blocker, in whom a head-up-tilt table test was performed after syncope of unknown origin. The test was chosen in order to reproduce the circumstances under which the syncope had happened. The monitoring device consisted of impedance cardiography as well as non-invasive beat-to-beat blood-pressure measurement. By these means it was possible to register a reduced peripheral vascular resistance after tilting the patient, a reduced cardiac output and bradycardia leading to syncope after four minutes of upright posture. This was suggestive for neurocardiogenic syncope as a cause for the spell experienced. Further non invasive diagnostic methods were normal. As the patient's family history was negative for syncopes, Torsade-de-pointes seemed unlikely.In this special case the non-invasive monitoring system of cardiac output, peripheral vascular resistance and beat-to-beat blood pressure measurements was useful as a supplemental tool during evaluation of syncope and helpful in decision-making against implantation of an ICD and for a more intense treatment with betablockers. Such monitoring devices offer new insights into the orthostatic regulation in young patients. PMID- 16258779 TI - Hypereosinophilic syndrome associated with acute necrotizing myocarditis and cardiomyopathy. AB - We report the rare case of a 55-year-old female with massive eosinophilic myocarditis and severe, however reversible, impairment of left ventricular function. The patient presented with reduced physical condition, progressive dyspnea on exertion and peripheral edema. The white blood count revealed a leukocytosis and markedly elevated peripheral blood eosinophilics (48.8%). An endomyocardial biopsy demonstrated massive myocardial infiltration with eosinophilic granulocytes and necrosis. The symptoms and laboratory parameters indicate the presence of a hypereosinophilic syndrome. The differential diagnosis of a Churg-Strauss syndrome is discussed. Medical heart failure treatment according to international guidelines and an immunosuppressive treatment with prednisolone (Decortin H) 1.5 mg/kgBW) were initiated. This therapy led to a dramatic reduction of the eosinophilic granulocyte count and normalization of the peripheral blood count, which correlated with a significant improvement of clinical symptoms. Consistently, an increase of left-ventricular function was observed. Upon successive dose reduction to a maintenance dosage of 10 mg prednisolone, the patient's clinical status and peripheral blood count remained stable. PMID- 16258782 TI - The heart in muscular dystrophies. PMID- 16258784 TI - [Recommendations for prevention and evidence-based medicine]. PMID- 16258785 TI - [Cost-effectiveness of prevention of coronary disease in Germany]. AB - It is generally accepted that the incidence of coronary heart disease can effectively be reduced by strengthening prevention. At the same time, it is still unclear how large the effects of life-style oriented preventive measurements such as diet and exercising are in everyday life. Furthermore, there is an ongoing debate on what measurements are effective. Thus, against the background of dwindling financial resources in health care the input of health economic evaluation is explicated. General issues of health economic evaluation are presented. After that, an overview on the current findings of cost-effectiveness in primary prevention of coronary heart disease is given. Risk factors are separately discussed. It is demonstrated that preventive measurements dealing especially with hypertension and hypercholesterolemia can be cost-effective. PMID- 16258781 TI - Current understanding of coronary in-stent restenosis. Pathophysiology, clinical presentation, diagnostic work-up, and management. AB - In-stent restenosis is the limiting entity following coronary stent implantation. It is associated with significant morbidity and cost and thus represents a major clinical and economical problem. Worldwide, approximately 250 000 in-stent restenotic lesions per year have to be dealt with. The pathophysiology of instent restenosis is multifactorial and comprises inflammation, smooth muscle cell migration and proliferation and extracellular matrix formation, all mediated by distinct molecular pathways. Instent restenosis has been recognised as very difficult to manage, with a repeat restenosis rate of 50% regardless of the mechanical angioplasty device used. Much more favourable results were reported for the adjunctive irradiation of the in-stent restenotic lesion, with a consistent reduction of the incidence of repeat in-stent restenosis by 50%. Data from the first clinical trials on drug-eluting stents for the treatment of in stent restenosis have shown very much promise yielding this strategy likely to become the treatment of choice. This review outlines the histological and molecular findings of the pathophysiology, the epidemiology, the predictors and the diagnostic work-up of in-stent restenosis and puts emphasis on the various treatment options for its prevention and therapy. PMID- 16258786 TI - [Primary prevention-psychosocial aspects]. AB - Psychosocial risk factors like low socio-economic status, social isolation and lack of social support, chronic work or family stress, depression, and hostility contribute significantly to the development and adverse outcome of coronary heart disease (CHD). The prevention of CHD should therefore include screening for psychosocial risk factors and adequate interventions. Methods to assess psychosocial risk factors are single item questions to be included in the cardiologist clinical interview and/or standardized questionnaires. Recommended interventions include an appropriate physician-patient interaction, multimodal, behavioral interventions with face to face or group counselling, and, in case of clinically significant depression, psychotherapy and medication. PMID- 16258787 TI - [Physical activity]. AB - There are a total of 30 publications regarding the role physical exercise in primary prevention of coronary artery disease, nearly a quarter of a million patients have participated in these studies. The average observation time reaches 10 years. According to the guidelines of the ACC/AHA Task Force this is the basis for the evidence/recommendation class I. The benefit derived from this treatment considerably exceeds the risk; thus, there is no need for additional studies. A great number of population-based risk factors were evaluated; in several randomized studies and meta-analyses concordant results were achieved with respect to direction and degree of the therapeutic effect (Level B). PMID- 16258788 TI - [Position paper on the primary prevention of cardiovascular diseases. Current position of the 25.3.2003 Statement of the Board of the German Society of Cardiology--heart and circulatory research work commissioned by for the board by Project Group on Prevention]. PMID- 16258789 TI - [Nutrition]. AB - The favorable effect of a healthy diet on the initial development and the further progression of atherosclerosis has been demonstrated. The effect is significant even in patients with adequate pharmacological treatment. Nutrition should be calorie-adjusted high in fiber, low in total fat and in saturated fats (<10% of calories) and cholesterol (<300 mg/day). Mono-unsaturated fatty acids and omega-3 fatty acids have beneficial effects. Therefore fatty fish more than once a week, nuts, fruits and vegetables (>400 g/day) have a prognostically favorable effect, whereas the supplemental use of antioxidative vitamins is of no proven benefit. These recommendations result from the experience gained over generations in millions of people in different countries (Mediterranean and Asian) with a low rate of coronary events and a high life expectancy. These dietary habits are associated with a low rate of neoplasm and of osteoporosis. The recommendation for this type of diet is a class-I recommendation. PMID- 16258790 TI - [Role of body weight for the prevention of coronary heart disease]. AB - The prevalence of obesity has increased considerably in recent years; only one third of our population displays a desirable, healthy weight. Persons with increased body fat mass are in a constant pro-inflammatory and pro-thrombotic condition; in addition they are often insulin-resistant und show cardiovascular risk factors. Therefore, obesity induces the development of atherosclerosis as an independent risk factor as well as a promoter for other risk factors. In addition, congestive heart failure is more frequent in obese subjects. By reducing body weight with a decrease of predominantly visceral fat all risk factors are improved as well as endothelial function. Effects on coronary atherosclerosis and congestive heart disease are until now insufficiently investigated. PMID- 16258791 TI - [Prevention of coronary heart disease: smoking]. AB - Smoking is the leading preventable cause of illness and premature death in Germany, claiming over 110,000 lives a year because it directly increases the risk of dying from heart disease, stroke, emphysema and a variety of cancers. The overwhelming majority of smokers begin tobacco use before they reach adulthood. Among those young people who smoke, the average age is now 13-14. In Germany, about 39% of male and 31% of female adults (age 18-60 years) continue to smoke, despite information about the unequivocally negative health consequences of smoking. The exact mechanisms of smoking-related vascular disease are not yet known. Smoking causes acute hemodynamic alterations such as increase in heart rate, systematic and coronary vascular resistance, myocardial contractility, and myocardial oxygen demand. These short-term effects could lower the ischemic threshold in smokers with coronary artery disease and contribute to the increased risk for acute cardiovascular events. Endothelial damage is thought to be an initiating event in atherosclerosis and early studies have demonstrated that long term smoking has direct toxic effects with structural changes of human endothelial cells. Recent research has shown the importance of the functional role of the endothelium in regulating vascular tone, platelet-endothelial interactions, leukocyte adhesion and smooth muscle cell proliferation via synthesis and release of a variety of substances such as nitric oxide. There is strong evidence that smoking leads to endothelial dysfunction mainly by increased inactivation of nitric oxide by oxygen-derived free radicals. Smoking also increases oxidative modification of LDL and is associated with lower HDL plasma levels. Smoking induces a systemic inflammatory response with increased leukocyte count and elevation of the C-reactive protein level. Importantly, the prothrombotic effects of smoking have been repeatedly demonstrated to cause alterations in platelet function, imbalance of antithrombotic vs prothrombotic factors, and decrease of fibrinolytic activity. Given the enormous health hazard of tobacco use, complete abstinence from smoking should be achieved. Smoking cessation counselling should be given to healthy subjects and even more vigorously to patients with manifested disease. Every effort should be undertaken to prevent children and adolescents from starting to smoke. Brief tobacco dependence treatment is effective, and every smoker should be offered at least brief treatment at every office visit. More intensive treatment is more effective in producing long-term abstinence from tobacco. Nicotine replacement therapy (nicotine patches or gum), clinician-delivered social support, and skills training are the three most effective components of smoking cessation treatment. A framework for tobacco control measures is necessary to reduce tobacco consumption and exposure to tobacco smoke. Recommendations on specific tobacco control interventions are: 1. increase in tobacco taxes; 2. comprehensive tobacco advertising bans; 3. legislation prohibiting smoking in work and public places; 4. prohibiting the sales of tobacco products to persons under 18; 5. comprehensive disclosure of the physical, chemical and design characteristics of all tobacco products; 6. training of health professionals to promote smoking prevention and cessation interventions; and 7. development of a national network of smoking cessation treatment services. PMID- 16258792 TI - [The role of lipid metabolism in the prevention of coronary heart disease]. AB - Prevention of cardiovascular disease should be considered as a continuum from low to high risk: those at the highest risk are patients with clinically manifest cardiovascular disease, followed by subjects without known cardiovascular disease at different levels of risk from high to low. Today there is clear evidence that an independent relationship exists between plasma LDL cholesterol levels and the risk for coronary heart disease. The relationship between other plasma lipoproteins and atherosclerosis is more complex. The threshold for individuals requiring LDL cholesterol reduction is determined by epidemiological data, randomized controlled trials, and economic considerations. Patients with familial dyslipidemia suffer early coronary morbidity and mortality. For these patients, consequent lowering of LDL cholesterol should be the primary objective. For patients with established coronary heart disease or other atherosclerotic disease and for those with diabetes, there is significant evidence that reducing LDL cholesterol, irrespective of the initial values, reduces the risk of further coronary events, stroke, and total mortality. For asymptomatic individuals, the treatment of plasma lipids should be based on their absolute coronary risk, including other cardiovascular risk factors. The goals for plasma LDL cholesterol have been set in national and international recommendations. The goals for LDL cholesterol in patients with low, moderate and high coronary risk are <160, <130 and 100 mg/dl, respectively. In some very high risk patients LDL level markedly below 100 mg/dl should be aimed at. HDL cholesterol and triglyceride measurements should be used to identify individuals at high multifactorial risk of cardiovascular disease and used as additional considerations in the selection of lifestyle and drug interventions. PMID- 16258793 TI - [Prevention of arteriosclerosis. Importance of the treatment of arterial hypertension]. AB - In most European countries and Northern America, cardiovascular diseases induced by atherosclerosis are the most common cause of death in older people. People surviving acute myocardial infarction or stroke suffer often by disabilities or handicaps. The lifelong care of such patients is expensive and plays a major role for increment of costs in public health systems. Prevention of atherosclerosis will reduce cardiovascular morbidity and mortality, enhance quality of life and prolong lifetime of patients. Therefore the worldwide accepted risk factors of atherosclerosis have to be treated consequently and early enough within the meaning of primary prevention. Hypertension is one of the six major cardiovascular risk factors and is defined as elevated blood pressure above 140/90 mmHg. In case of hypertension, diagnostic efforts has to be focussed on detection of additional cardiovascular risk factors, secondary forms of hypertension, end organ damage or associated diseases. All therapeutic strategies are based on life style changes, which cover weight reduction, sodium restriction, controlled alcohol consumption and increment in physical activity. Pharmacotherapy will be added in regard to the global risk of the patient and the success of the life style changes. Selection of antihypertensives and their optimal combination will be determined by associated diseases (compelling indication), side effects and individual response in blood pressure. Goal of treatment is the normalization of blood pressure below 140/90 mmHg independent of age or sex. In diabetics and in case of nephropathy the goal is set lower (below 130/80 mmHg).There is strong evidence that reduction in blood pressure is followed by a decrease in the incidence of myocardial infarction, stroke, heart failure, nephropathy, and even in cardiovascular mortality. The success of antihypertensive therapy is greater in high risk patients like older people, patients with isolated systolic hypertension or diabetics. Risk reduction correlates well with the degree in blood pressure reduction. However, to minimize cardiovascular risk in hypertensives all additional risk factors have to be treated too. PMID- 16258794 TI - [Total risk for cardiovascular disease. At what point is medical prophylactic medication useful?]. AB - Despite the epidemiological importance of coronary artery disease: cardiovascular events are rare from the individual viewpoint. There is considerable uncertainty when to start medical treatment. A given risk factor modification results in a relative risk reduction independent of the global risk. Therefore the global risk determines the absolute benefit of a preventive measure. The global risk can be estimated using different scoring systems. Using the global risk and the expected relative risk reduction, the number needed to treat (NNT) to avoid one event or cardiac death can be calculated. The NNT is a measure for the usefulness of a preventive intervention. A NNT of <200 appears acceptable for primary prevention. This can be achieved with pharmacological preventive strategies if the global risk of 10 years is > or =20%. As age is one of the most important risk predictors the need for treatment at comparable risk factor constellations is age dependent. Risk stratification with estimation of the NNT is therefore important for the decision to treat or not to treat. PMID- 16258795 TI - [Primary prevention of coronary heart disease with aspirin]. AB - According to meta-analysis and the results of the two studies with the highest power, aspirin is effective in primary prevention of coronary heart disease. These beneficial effects, however, are at least partially out-weight by unwanted effects-such as intense gastrointestinal bleeding and hemorrhagic stroke. These side effects remain constant with increasing risk of coronary heart disease, whereas the protective effects increase. If an annual risk of coronary heart disease of < or =0.6% exists, aspirin is normally not indicated; for a risk of 0.7-1.4% the facts should be discussed with the patient. If a risk of > or =1.5% exists, aspirin should be given. Problems of aspirin therapy--such as "aspirin paradox" and "aspirin resistance"--have been documented for secondary prevention; they might, however, have likewise clinical implications in primary prevention. PMID- 16258796 TI - [Role of hormone-replacement therapy for prevention of coronary artery disease in women]. AB - The postmenopausal increase in the incidence of coronary artery disease implied a protective effect of estrogens. Nonrandomized, clinical and experimental studies have supported this notion. In the first randomized study (HERS 1998) no protective effect on prognosis of postmenopausal women with coronary artery disease was demonstrated. Also, in healthy postmenopausal women no beneficial effect of a hormone-replacement therapy on coronary events was shown (WHI-Study 2002, 2004). Therefore, hormone-replacement therapy is not recommended for prophylaxis of cardiovascular disease in healthy women or in women with documented coronary artery disease (Recommendation class I, evidence-level A). The continuation or the start of a hormone- replacement therapy is only justified for therapy of severe menopausal symptoms. Women should be informed that changes in lifestyle including not smoking, a heart healthy diet, and regular exercise are the most important measures to prevent cardiovascular diseases. PMID- 16258797 TI - [Value of coronary artery calcium measurements in primary prevention]. AB - Frequently, myocardial infarction or sudden coronary death are the index manifestations of coronary artery disease. In view of the high out-of-hospital mortality of acute myocardial infarction, medical care is unable to provide a benefit for many patients. Against this background, it is an important aim of measuring coronary calcium to identify asymptomatic subjects with an increased coronary risk who are likely to derive a benefit from risk-modifying therapy. Coronary calcium is a largely specific expression of coronary atherosclerosis and is correlated with overall coronary plaque volume. Due to the complex biology of the vessel wall and its ability to undergo compensatory remodelling, coronary calcium does not necessarily indicate significant stenosis. Coronary calcium is found in 70-80% of plaque ruptures but only in a minority of plaque erosions. It neither indicates a "vulnerable" nor a "stable" plaque. Six independent studies including healthy self-referred and physician-referred volunteers consistently describe the predictive value of coronary calcium with regard to coronary and cardiovascular clinical events. After adjusting for coronary risk factors, increased amounts of coronary calcium are associated with a 5- to 10-times elevated relative risk. Only recently have the first results from strictly unselected, population-based cohorts been reported which confirm the predictive ability of coronary calcium measurements. Concordant with actual guidelines issued by US-American and European expert panels, coronary calcium measurements can be used especially in patients with an indeterminate risk on the basis of clinical assessment and risk factor analysis. Substantially elevated coronary calcium scores provide a rationale for intensified risk-modifying therapy. This is also true for elderly patients in whom the established risk factors lose some of their predictive power. The use of coronary calcium measurements in self referred patients or as a primary means for risk stratification is not encouraged. PMID- 16258798 TI - [Glucose metabolism]. AB - The markedly increased peri-interventional risk (PCI and CABG) in patients with type-2 diabetes mellitus may be reduced by adjusting blood glucose values to a near-normal level. This adjustment should be realized acutely by glucose-insulin potassium infusions. In long-term therapy, the target value should be achieved independent of the pharmacological principle of blood glucose reduction. Among the available oral antidiabetic agents, metformin, acarbose and glitazones seem to be cardioprotective via pleotropic effects. Given an optimal stent implantation and administration of GP IIb/IIIa inhibitors during coronary interventions, results are similar to those of non-diabetics. PMID- 16258801 TI - Nomenclature of hepatic anatomy and resections: a review of the Brisbane 2000 system. AB - The Brisbane 2000 system of nomenclature of hepatic anatomy and resections was introduced to provide a universal terminology in an area that was plagued by confusing and inappropriate terminology. The article describes historical developments central to the emergence of the new terminology and describes the terminology, its attributes, and rules of application. PMID- 16258799 TI - [Primary prevention of coronary heart disease? What is cost effective in the clinical practice?]. AB - In the primary prevention of coronary heart disease (CHD), the effect of aspirin and statins is well documented in several controlled randomized trials. For aspirin the results can be transferred into clinical practice due to its low price; for the more expensive statins, however, serious economic problems exist. In contrast to secondary prevention these drugs do not reach cost-efficiency in primary prevention; due to their high prices for the criteria of the randomized controlled studies values >60 000 or >100 000 [US dollars/YLS] are gained. Data from England and Scotland indicate that according to the inclusion criteria of the WOSCOPS- and AFCAPS/TexCAPS studies almost 20 and 60%, respectively, of the adult population had to be treated with a statin. Results of newer studies may even increase these numbers. These costs cannot be covered by any health care system. Primary prevention of CHD with statins reveals paradigmatically that for financial reasons evidence-based medicine can no longer be transferred into clinical practice. The limited resources of all health care systems make rationing with treatment allocation only to the high risk groups necessary. The American, European and German guidelines propose a > or =2% annual risk of CHD as the limit, for financial reasons the Britisch recommendations favor a limit of 3%; in order to save >50% of the costs. Despite the financial restraints of the German health care system, the limit of > or =2% annual risk of CHD as proposed by the German Cardiac Society may be realistic when the different preventive measures are applied following a step-by-step plan based on the costs. According to the Procam algorithms, persons without diabetes mellitus or familiar disposition, who in case of nicotine abuse have given up smoking and if hypertensive have blood pressure values within the therapeutic range, statins are only to be given under the following conditions: LDL-cholesterol > or =175 or > or =190 mg/dl, for a HDL-cholesterol < or =35 or < or =45 mg/dl, or triglyceride levels > or =200 or > or =175 mg/dl, respectively. Diabetics without CHD have the same risk as non-diabetics with CHD. Therefore, in diabetics the same measures should be taken for primary prevention as in non-diabetics for secondary prevention. Evaluation of cost-efficiency indicates that intensive blood sugar control as well as intensive antihypertensive treatment and application of statins are all cost-effective in primary prevention of diabetics. PMID- 16258802 TI - Anterior approach for major right hepatic resection. AB - We review the history, indications, and latest modifications of the surgical technique of the anterior approach for right hepatectomy for massive tumors. The anterior approach provides a "no-touch" technique in resecting large right-lobe tumors, reduces bleeding volume, decreases the chance of iatrogenic rupture of tumors, and probably prolongs survival. PMID- 16258803 TI - Systematic hepatectomy for liver cancer. PMID- 16258805 TI - Step classification is useful for the determination of indications for systematized hepatectomy in hepatocellular carcinoma. AB - BACKGROUND/PURPOSE: The effectiveness of systematized hepatectomy with transection of Glisson's pedicle at the hepatic hilus has not been clarified in detail in relation to previous staging systems. Outcomes after systematized hepatectomy in patients with hepatocellular carcinoma (HCC) were examined in relation to our new staging system. METHODS: We retrospectively studied 955 patients with HCC who underwent hepatectomy from 1989 through 2002. We classified patients with HCC into four groups according to the pathological findings (pathological step [p-step]): p-step 1, HCC with absence of vascular invasion and absence of intrahepatic metastasis; p-step 2, HCC with vascular invasion and/or intrahepatic metastasis; p-step 3, HCC with major vascular invasion and/or intrahepatic metastasis to both lobes of the liver; and p-step 4, HCC with distant metastasis, including lymph node metastasis or ruptured HCC). We separated the liver into three segments (Takasaki's liver segments). Systematized hepatectomy was classified as systematized segmentectomy or larger resection, and partial segmentectomy. Segmentectomy refers to resection of one of Takasaki's segments. RESULTS: Systematized segmentectomy did not affect recurrence-free survival, by univariate analysis, in patients with p-step 1, p-step 3, or p-step 4. However, systematized segmentectomy or larger resection was significantly associated with patient recurrence-free survival, by univariate analysis, in patients with p-step 2. Multivariate analysis also showed systematized segmentectomy or larger resection as a significant independent prognostic factor in patients with p-step 2. CONCLUSIONS: Systematized segmentectomy is suitable for patients with p-step 2 HCC according to this step classification. PMID- 16258804 TI - Systematic hepatectomy for small hepatocellular carcinoma in Korea. AB - BACKGROUND/PURPOSE: Systematic hepatectomy for small hepatocellular carcinoma (HCC) is a widely preferred modality, but evidence concerning its benefits is lacking. The aim of this study was to document hepatic resection for small HCC in Korea, and to determine whether patient survival or the pattern of tumor recurrence was influenced by the methods used. METHODS: Ten major hospitals that perform hepatectomy for HCC in Korea were surveyed for surgeons' opinions concerning systematic hepatectomy and current trends in hepatic resection for small HCC. An analysis was also performed of 119 patients who underwent curative hepatectomy for small HCC (size < 5 cm) between January 2000 and December 2002 at Seoul National University Hospital. Seventy-four of these 119 patients underwent anatomical resection (AR) and 45 had a non-anatomical resection (NAR). Recurrence free survival, recurrence pattern, overall survival rates, and the risk factors for recurrence were analyzed. RESULTS: In the survey, eight of ten surgeons preferred systematic hepatectomy and considered it to aid prognosis. No significant difference was found between the AR and NAR groups in terms of the clinicopathologic findings, except that the presence of underlying hepatic cirrhosis was more prevalent in the NAR group. The postoperative morbidity rate was higher in the NAR group (33.3% vs 27.0%), but this difference was not statistically significant. The respective 1- and 3-year recurrence-free survival rates were 78.1% and 49.7% in the AR group, and 68.9% and 46.5% in the NAR group (P > 0.05). The corresponding 1- and 3-year overall survival rates were 88.8% and 80.8% in the AR group and 91.0% and 71.4% in the NAR group (P > 0.05). CONCLUSIONS: Although systematic hepatectomy seems to be superior to non anatomical hepatectomy from the oncological and anatomical aspects, this superiority is not reflected by the recurrence patterns or the survival and recurrence rates of the two procedures. Postoperative recurrence appears, rather, to be related to the underlying liver condition. PMID- 16258806 TI - Hepatocyte transplantation for total liver repopulation. AB - Hepatocyte transplantation (HT) is an attractive therapeutic alternative to liver transplantation. A number of experiments have shown the feasibility of total liver parenchymal cell replacement by transplanted hepatocytes. In this review, we would like to highlight researches and clinical reports of HT for liver repopulation. Cellular source of clinical HT should be safety. Immortalized cells, hepatic stem cells, and other stem cells have been used for an experimental model for HT. The exact mechanism of the cell engraftment after HT has not been completely understood, although there were some markers to detect and investigate transplanted cells. In order to achieve liver repopulation following HT, a mild hepatic damage may need to facilitate cell engraftment and replace the host liver by transplanted cells. Hormonal factor may use for the same purpose. Despite the results of preclinical studies promising clinical benefits for cell therapy, the clinical experience of HT has been disappointing, except in a few cases. HT may become an alternative for liver transplantation in the future; however, many efforts should made before establishing an effective method for HT and liver replacement therapy. PMID- 16258807 TI - Placement of a double-pigtail ERBD tube after primary closure of the common bile duct in three patients with choledocholithiasis. AB - We devised a procedure for the placement of a double-pigtail endoscopic retrograde biliary drainage (ERBD) tube as an alternative to the placement of a T tube. We used the procedure, after primary closure of the common bile duct, in three patients undergoing surgery for choledocholithiasis. All three patients were in their eighties, and all were diagnosed with cholecystolithiasis and choledocholithiasis. In all three, there were concerns about possible complications with the use of a T-tube. Two of the patients were senile and were thought to be likely to pull out the tube, and incomplete fistulation was considered possible in the third patient. Thus, there was an increased risk of bile peritonitis in all three patients. Placement of the ERBD tube was successful in all three patients, and there were no postoperative complications. The hospital stay was a few weeks shorter than the usual stay with the placement of a T-tube. We conclude that primary closure of the common bile duct with the placement of a double-pigtail ERBD tube is clinically safe and advantageous for choledocholithiasis patients with senile dementia and for patients with possible incomplete fistulation. PMID- 16258808 TI - Unsuspected gallbladder carcinoma after laparoscopic cholecystectomy. AB - BACKGROUND/PURPOSE: Many cases have been reported of disastrous port-site recurrence after laparoscopic cholecystectomy (LC) revealed unsuspected gallbladder carcinoma (GBC). Some investigators have reported that the prognosis of patients after LC showed unsuspected GBC is not worsened by laparoscopic procedures. We retrospectively reviewed our cases and the literature to reconfirm the intrinsic risks of LC for unsuspected GBC. METHODS: Of 1663 patients who underwent LC from January 1991 to December 2003 in a single institution, 9 (0.54%) with unsuspected GBC were reviewed. RESULTS: These 9 patients consisted of 5 men and 4 women, whose ages ranged from 58 to 87 years, with a median age of 73 years. Two patients with a pT1a tumor (limited to mucosa) and 2 patients with a pT1b tumor (muscle layer) underwent no further operation. The remaining 5 patients with a pT2 tumor (subserosa) underwent further operations with lymph node dissection. Five patients (2 patients with pT1b and 3 patients with pT2) developed recurrence and all of them died within a median period of 19 months (range 14-37 months) after LC. The causes of death were bone metastases in 1 patient (pT2), local recurrence in 2 patients (pT1b and pT2), and peritoneal metastasis in 2 patients (one elderly patient with pT1b who underwent laparoscopic common bile duct exploration, and one patient with pT2 in whom the cystic duct was damaged during surgery). Four patients (2 with pT1 and 2 with pT2) have been doing well with a median follow-up of 39.5 months (range 12-99 months) after LC. CONCLUSIONS: Surgeons should always prevent bile spillage during LC and when removing the resected gallbladder. When laparoscopic common bile duct exploration is planned, especially for elderly women, surgeons should also bear in mind the increasing possibility of unsuspected GBC. PMID- 16258809 TI - Risk factors for early death after liver resection in patients with solitary hepatocellular carcinoma. AB - BACKGROUND/PURPOSE: Although most patients who receive hepatectomy for a solitary hepatocellular carcinoma (HCC) have a relatively fair result, some have a poor prognosis. The aim of this study was to evaluate the risk factors for early death after hepatectomy in patients with a solitary HCC. METHODS: Eligible patients (n=110) who had undergone hepatectomy for solitary HCC between 1990 and 2002 and were able to be followed up for more than 2 years after the hepatectomy were divided into two groups, those who died of cancer recurrence within 2 years (early-death group; n=18) and those who survived for more than 2 years after the surgery (survival group; n=92). Risk factors for early death after liver resection were evaluated by univariate and multivariate analyses. RESULTS: The gross tumor classification, tumor diameter, macroscopic portal vein invasion, microscopic growth pattern, microscopic vascular invasion (MVI), and the width of the surgical margin were significant (P<0.05) factors by univariate analysis. Multivariate analysis showed that the presence of MVI was an independent and significant risk factor for early death of recurrence. CONCLUSIONS: Among patients with solitary HCC, the presence of MVI indicates a poor prognosis. These patients need adjuvant chemotherapy in the early period after hepatectomy. PMID- 16258810 TI - Gallbladder lymphangioma: a case report and review of the literature. AB - Lymphangiomas are benign neoplasms usually occurring in childhood and located in the head and neck. Intraabdominal lymphangiomas account for less than 5% of cases. The involvement of the gallbladder is rare. We report a case of a 29-year old woman who presented with right upper quadrant pain that had persisted for 6 months. Imaging with ultrasound and magnetic resonance imaging (MRI) revealed a multiseptated lesion surrounding the gallbladder. The patient underwent an exploratory laporatomy, and the mass was resected en bloc with the gallbladder. Histological evaluation of the cystic mass revealed findings consistent with lymphangioma. The prognosis is generally good after complete surgical excision, as was the case for our patient. PMID- 16258811 TI - Spontaneous rupture of a hepatic cystadenoma and cystadenocarcinoma: report of two cases. AB - Hepatobiliary cystadenomas and cystadenocarcinomas are rare tumors. Differentiating between these tumors and benign hepatic cysts may be difficult. Because of their rarity, diagnosis is often delayed and may result in inaccurate treatment, resulting in unnecessary morbidity and mortality. The purpose of this report is to draw attention to these entities and their complications. We report on two cases with spontaneous rupture of hepatobiliary cystadenoma and cystadenocarcinoma cysts, initially treated as simple hepatic cysts by aspiration, or by aspiration combined with sclerotherapy. The spontaneous rupture of the cysts appeared years after the initial treatment of the cysts, leading in one case to a prolonged stay in an intensive care unit. In both cases, a formal liver resection was carried out and microscopic investigations revealed a mucinous cystadenocarcinoma and cystadenoma. In conclusion, although hepatobiliary cystadenomas and cystadenocarcinomas are rare findings, they should not be forgotten in the diagnostic workshop when examining patients with hepatic cysts. If hepatobiliary cystadenomas and cystadenocarcinomas cannot be excluded following radiological imaging, surgery is recommended. PMID- 16258812 TI - Metastatic colon cancer from intrahepatic cholangiocarcinoma. AB - A 62-year-old man had been followed because of an elevated serum level of carcinoembryonic antigen without the detection of any cancer lesions. However, there was a sudden increase in the serum level of carcinoembryonic antigen, and abdominal imagings showed a hepatic tumor with peripheral intrahepatic bile duct dilatation, and a submucosal tumor at the sigmoid colon with intact mucosa. Histopathological findings showed that the hepatic tumor had perineural invasion, suggesting an intrahepatic cholangiocarcinoma, and that the colon tumor infiltrated the submucosa, while its mucosa was intact. Both tumors showed similar pathological features and were positive for cytokeratin 20 and 7. These findings suggested intrahepatic cholangiocarcinoma with metastatic sigmoid colon cancer. PMID- 16258813 TI - Cystic duct directly joining the main pancreatic duct. AB - A unique anomaly of the direct union between the cystic duct and the main pancreatic duct is presented. A 19-year-old man with a history of repeated epigastralgia underwent endoscopic retrograde cholangiopancreaticography that showed a direct union between the cystic duct and the main pancreatic duct. No pancreaticobiliary maljunction was noticed. Cholecystectomy accompanied by resection of the long cystic duct was performed. The excised gallbladder showed cholesterolosis, chronic cholecystitis, and hyperplasia of the pseudopyloric glands microscopically. The patient has been well for 3 years since surgery. PMID- 16258814 TI - The usefulness of early laparoscopic surgery for determining a medical treatment plan in severe acute pancreatitis. AB - A 41-year-old woman presented with sudden onset severe epigastric pain. She was diagnosed as having severe acute pancreatitis (SAP) with an APACHE II score of 10, and initially, general supportive therapy was performed. Because the patient's condition kept worsening, early laparoscopic surgery was performed. It revealed swelling, hemorrhagic necrosis of the pancreas, and massive fluid collection in the abdominal cavity. Kocherization and bursectomy were performed for these lesions, and drainage tubes were indwelled. The white blood cell count (WBC) and serum interleukin-6 value prominently improved shortly after the operation, and the WBC count remained at a low level. The patient gradually recovered and was finally discharged 39 days after her initial presentation. To summarize, we performed a successful exploration for SAP, and we found that postoperative continuous lavage effectively eliminated toxic enzymatic exudates in the abdominal cavity and prevent them from circulating. Regarding SAP with peritonitis, early laparoscopic surgery is very useful for determining the pathological extent of the disease and for selecting appropriate treatment options. PMID- 16258815 TI - Bacteriophage therapy: a revitalized therapy against bacterial infectious diseases. AB - Bacteriophage (phage) therapy involves using phages or their products as bioagents for the treatment or prophylaxis of bacterial infectious diseases. Much evidence in support of the effectiveness of phage therapy against bacterial infectious diseases has accumulated since 1980 from animal model studies conducted in Western countries. Reports indicate that appropriate administration of living phages can be used to treat lethal infectious diseases caused by gram negative bacteria, such as Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii, Klebsiella pneumoniae, Vibrio vulnificus, and Salmonella spp., and gram-positive bacteria, such as Enterococcus faecium and Staphylococcus aureus. The phage display system and genetically modified nonreplicating phages are also effective for treatment of Helicobacter pylori and P. aeruginosa, respectively. In addition to phage particles per se, purified phage-encoded peptidoglycan hydrolase (lysin) is also reported to be effective for the treatment of bacterial infectious diseases caused by gram-positive bacteria such as Streptococcus pyogenes, S. pneumoniae, Bacillus anthracis, and group B streptococci. All phage lysins that have been studied to date exhibit immediate and strong bacteriolytic activity when applied exogenously. Furthermore, phage coded inhibitors of peptidoglycan synthesis (protein antibiotics), search methods for novel antibacterial agents using phage genome informatics, and vaccines utilizing phages or their products are being developed. Phage therapy will compensate for unavoidable complications of chemotherapy such as the appearance of multidrug resistance or substituted microbism. PMID- 16258816 TI - Feasibility of flow cytometry for the detection of bacteria from body fluid samples. AB - The detection of microorganisms in body fluids is indispensable for identifying the source of infection and is one of the important examinations that influence subsequent treatment. In order to quickly detect bacteria in body fluid samples, a flow cytometry-based experimental automated bacteria counter (BF-FCM), was tested to determine its clinical value. The results for detectability obtained with the BF-FCM were compared with those obtained by conventional culture and Gram-staining techniques. We evaluated a total of 318 body fluid samples, excluding bile samples from which fungus alone was isolated. The samples consisted of 176 bile, 64 ascites, 42 pleural fluid, and 36 cerebrospinal fluid samples. Among the 318 samples, 154 (48.4%) were culture-positive. Of these 154, the BF-FCM identified 130 as positive (sensitivity, 84.4%). Of the 164 samples that were culture-negative, 141 were negative by the BF-FCM (specificity, 86.0%). Based on the culture results, the BF-FCM detected bacteria with a positive predictive value of 85.0% (130 of 153 samples), a negative predictive value of 85.5% (141 of 165 samples), and percent agreement of 85.2%. Although there were 23/164 (14.0%) false-positive samples, we consider that the BF-FCM, in combination with Gram staining and conventional cultures, would be helpful in the diagnosis and management of patients with diseases such as bacterial meningitis that present emergently. PMID- 16258817 TI - Presence of a mutation in ponA1 of Neisseria gonorrhoeae in numerous clinical samples resistant to various beta-lactams and other, structurally unrelated, antimicrobials. AB - The number of resistant strains in patients with Neisseria gonorrhoeae urethritis has been increasing, making effective treatment difficult. Chromosomally mediated penicillin-resistant N. gonorrhoeae arise through alterations in penicillin binding proteins (PBPs) and a decrease in outer membrane permeability. To understand the occurrence of penicillin resistance in patients with N. gonorrhoeae infection, we performed this study. In addition, we studied minimum inhibitory concentrations (MICs) of antimicrobials against N. gonorrhoeae strains. We measured the MICs of penicillin G, other beta-lactams, and other kinds of antimicrobials against 53 clinical N. gonorrhoeae isolates from male patients with urethritis in Hyogo and Osaka, Japan. The ponA genes, encoding PBP 1 of these isolates, were sequenced. Of the 53 isolates tested, 41 strains showed some resistance to penicillin G. A mutation in the ponA (ponA1) gene was identified in 46 isolates. There was a tendency that ponA mutant (ponA1) in N. gonorrhoeae led to higher antimicrobial MICs of beta-lactam antimicrobial agents (including penicillin) than those of non-ponA mutants. However, we found lower than expected MICs of penicillin and beta-lactams even in ponA mutants. Therefore, we consider that detailed investigations for the further understanding of the effect of other genes, such as penC (which is reported to be related to ponA1 in achieving high-level penicillin resistance) should be our next step. PMID- 16258818 TI - Unselective use of intranasal mupirocin ointment for controlling propagation of methicillin-resistant Staphylococcus aureus in a thoracic surgery ward. AB - We executed a blanket-use program of intranasal application of mupirocin ointment to control the propagation of methicillin-resistant Staphylococcus aureus (MRSA) that occurred in a thoracic surgery ward of a university hospital. During an intervention of 14 weeks, all patients admitted to the ward for scheduled surgery received the ointment to their nares three times daily for 3 days before surgery, once on return to the ward, and three times weekly for the following 2 weeks. None of 84 patients was newly colonized with MRSA, and the daily rates of patients with MRSA in a recovery room in the ward significantly decreased in the period. We consider that the unselective application of mupirocin ointment is one of the effective measures to control MRSA propagation in a thoracic surgery unit. PMID- 16258819 TI - Evaluation of a newly identified soluble CD14 subtype as a marker for sepsis. AB - CD14, a high-affinity receptor for lipopolysaccharide (LPS), is a glycoprotein expressed on the surface membranes of monocytes/macrophages. We have identified a previously unknown form of soluble CD14, named soluble CD14 subtype (sCD14-ST), that is increased in patients with sepsis. To measure sCD14-ST concentrations in plasma, we prepared anti-sCD14-ST antibodies and developed an enzyme immunoassay (EIA) for this soluble form of CD14. With this assay, quantitative measurements are available within 4 h, and we compared the levels of sCD14-ST in plasma from normal subjects (healthy controls), patients with systemic inflammatory response syndrome (SIRS), and sepsis patients. The level of sCD14-ST in subjects with sepsis was much higher than the levels in subjects with SIRS and the healthy controls. Additionally, when a subject's sCD14-ST level was used as a diagnostic marker for sepsis, the area under the receiver operating characteristic (ROC) curve was 0.817, thereby demonstrating that elevated sCD14-ST levels were a better marker for sepsis than the other molecular markers we tested. sCD14-ST levels also correlated with procalcitonin (PCT) levels and with sequential organ failure assessment (SOFA) scores. Finally, changes in sCD14-ST concentration correlated with the severity of sepsis. Taken together, these results indicate that sCD14-ST is a useful marker for the rapid diagnosis of sepsis and for monitoring the severity of the disease. PMID- 16258820 TI - Surgical antimicrobial prophylaxis in transurethral ureterolithotripsy. AB - The aim of this study was to establish a standard protocol for surgical antimicrobial agents for patients who received transurethral ureterolithotripsy (TUL). We retrospectively reviewed the medical charts of patients who received TUL. From October 2002 to December 2003, 2 days (total, four times) of antimicrobial prophylaxis (AMP) was done, and from January 2004 to December 2004, single prophylaxis was done. We analyzed the incidence of postoperative fever and other factors associated with TUL and compared these factors between the 2-day AMP and single-AMP groups. Of 135 patients with TUL, 66 patients were in the single-AMP group and 69 in the 2-day-AMP group. The incidences of postoperative fever were 4.5% in the single-AMP group and 11.6% in the 2-day-AMP group. No statistically significant difference was found in the incidence of postoperative fever between the two groups. Our study showed that single AMP was effective for patients receiving a TUL operation. In the future, a prospective randomized study will be needed for a large series, with various types of lithotripters being included. PMID- 16258821 TI - Legionella pneumophila infection in the Taiwan area. AB - The aim of this study was to explore the epidemiological distribution of legionellosis among pneumonia patients in Taiwan. From January 2001 to December 2003, specimens (i.e., sputum, urine, and serum) from a total of 5097 patients with pneumonia or pneumonia-like disease registered at hospitals in the Taiwan area were analyzed for possible Legionella infection. Following the guideline issued by the Centers for Disease Control and Prevention (CDC) in the United States, a total of 237 pneumonia patients were diagnosed with legionellosis, with an incidence rate among pneumonia patients in this area of 4.7% (237/5097). The paired-serum antibody test was found to be the most effective detection method, followed by urine-antigen detection and the sputum culture method. Analysis of distribution showed that: (1) male and female occurrence rates were 70.9% (168/237) and 29.1% (69/237), respectively; (2) occurrence rates in different age groups, i.e., those aged between 61 and 80 years, those aged between 41 and 60, and those aged between 21 and 40 were 50.2% (119/237), 26.2% (62/237), and 12.2% (29/237), respectively; (3) autumn was the peak season for infection, followed by winter, summer, and spring, sequentially. This is the first study in Taiwan to have followed the three-method guideline issued by the CDC and it is the second report in Taiwan involving the investigation of a large series of pneumonia patients for legionellosis detection. PMID- 16258822 TI - A patient with seminal vesiculitis prior to acute chlamydial epididymitis. AB - This is the first report of a case of seminal vesiculitis prior to acute chlamydial epididymitis. At the first visit to the clinic, the patient wished to check whether he had Chlamydia trachomatis in his genital tract, because his wife had been diagnosed as having chlamydial cervicitis. He had no specific symptoms at that time; however, transrectal ultrasonograpy (TRUS) revealed swelling of seminal vesicles, which suggested the presence of seminal vesiculitis. Two days after the first visit, he had high-grade fever and was diagnosed as having acute epididymitis caused by C. trachomatis. We had previously reported that seminal vesiculitis was always complicated with acute epididymitis, so this case could provide important evidence that seminal vesiculitis might precede acute epididymitis. It suggested that acute epididymitis could be affected by seminal vesiculitis via the retrograde transmission route. PMID- 16258824 TI - Serum antibody response to tuberculosis-associated glycolipid antigen after BCG vaccination in adults. AB - Tuberculous glycolipid antigens (TBGLs) are derived from the cell walls of Mycobacterium tuberculosis. Detection of anti-TBGL antibody in serum has recently become possible for the serodiagnosis of active tuberculosis. TBGL is not indigenous to M. tuberculosis, but it is widely found in Mycobacterium species. To elucidate the influence of M. bovis bacille Calinette-Guerin (BCG) vaccination, we assayed serum anti-TBGL antibody after BCG vaccination in adults. BCG vaccination was done for 20 Japanese healthcare workers with a negative tuberculin skin test reaction, and serum was collected 0, 2, 4, and 8 weeks and 1 year after vaccination. The tuberculin skin test became positive in 85% of the subjects. The mean anti-TBGL antibody titer remained negative throughout the observation period, but was elevated significantly compared with the pre vaccination level, peaking at week 4 and showing a reduced level 1 year post vaccination. These results showed that serological diagnosis using anti-TBGL antibody was not influenced by prior BCG vaccination. PMID- 16258823 TI - Antimicrobial activities of tosufloxacin against Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella branhamella catarrhalis isolated from otolaryngological infectious diseases. AB - In 2003, the Japan Society for Infectious Diseases in Otolaryngology conducted its third nationwide survey of clinical isolates from otolaryngological infectious diseases. We selected three primary causative organisms of otolaryngological infectious diseases, Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella Branhamella catarrhalis, and evaluated their sensitivities to tosufloxacin (TFLX), a new oral quinolone, because the survey revealed a rise in drug-resistant strains, suggesting potential problems with the antibiotics commonly used against these organisms. The minimum inhibitory concentration (MIC)(90) values of TFLX against S. pneumoniae, H. influenzae, and M. catarrhalis were 0.25 microg/ml, 45 degrees in 76.6% of patients. Preoperative spondylolisthesis was found in 48% on radiological studies. All the patients were treated surgically with resection of the cyst. No fusion was performed as a first line procedure. However subsequent fusion was necessary in one patient who developed symptomatic spondylolisthesis. Mean follow-up period was of 45 months ranging from 18 to 105 months. Only one recurrence occurred during the follow-up period. An excellent or good functional outcome was seen in 97.4% of cases, and 89% of the patients with motor deficit recovered. CONCLUSIONS: Surgical resection of lumbar synovial cysts is an effective treatment associated with very low morbidity. Synovial cysts are associated with increased grade and frequency of facet joint asteoarthritis but not with increased grade or frequency of degenerative disc disease compared with patients without cysts. In the author's opinion, at the present time, there is no reliable criterion which allows the development of a symptomatic spinal instability to be predicted in patients with a preoperative spondylolisthesis and therefore fusion as a first line procedure is still debatable. PMID- 16258840 TI - The use of ciliary neurotrophic factor to promote recovery after peripheral nerve injury by delivering it at the site of the cell body. AB - Despite a body of evidence showing that various neurotrophic factors support the survival of nerve cells and stimulate axonal outgrowth, doubt remains about their optimal site of application as well as the more compelling question of what clinical benefit, if any, they would confer. Ciliary neurotrophic factor (CNTF) supports the survival of motorneurons in vitro and in vivo. Direct delivery of CNTF to the cell bodies may help reduce the side effects and overcome the problem of rapid systemic clearance. The aim of this study was to establish whether nerve regeneration may be improved upon by the controlled addition of a specific humoral neurotrophic substance (CNTF) at the level of the cell body. Three groups of five adult sheep were used. The first group acted as normal controls. In the second and third groups, the median nerve was divided and repaired using an epineurial suture technique. In the second group, CNTF was supplied into the CSF at the level of C6 by an implanted osmotic pump. In the third group physiological saline was placed in the osmotic pump. The animals underwent comprehensive electrophysiological and isometric tension experiments at six months. All of the animals had reduced electrophysiological, morphometric and isometric tension indices after surgery compared to normal. The CNTF group had better results than the saline group in the following; (1) area and amplitude of the muscle action potential (2) the percentage of tetanus and muscle mass preserved after repair. These differences were only statistically significant for amplitude of the muscle action potential. No statistical difference was found in the morphological indices (fibre diameter, axon diameter, myelin thickness and internodal length) between the CNTF and saline groups. CNTF does not confer a functional benefit when applied at the level of the cell body. PMID- 16258841 TI - Neuroprotective effects of indomethacin and aminoguanidine in the newborn rats with hypoxic-ischemic cerebral injury. AB - Nitric oxide (NO) and prostaglandins (PG) play important roles in delayed mechanisms of brain injury. While NO disrupts oxidative metabolism, prostaglandins are responsible for free radical attack in reperfusion interval. Relatively little is known about neuroprotection exerted at this level in perinatal models. The aim of this study was to investigate the effect of indomethacin and aminoguanidine on endogenous inducible nitric oxide synthase (iNOS) biosynthesis and neuroprotection in the newborn rats with hypoxic ischemic cerebral injury.Seven-day old rat pups with model of hypoxic-ischemic cerebral injury were randomly divided into four study groups. Group C (n=18; served as a control) pups were given physiologic saline (SF). Group I (n=18) pups were treated with indomethacin at a dose of 0,2 mg/kg per 12 h. Group A (n=20) pups were treated with aminoguanidine at a dose of 300 mg/kg per 8 h. Administration of drugs and SF were begun half an hour after hypoxic-ischemic insult in these groups. Group I+A (n=18) pups were treated with indomethacin at a single dose of 0.2 mg/kg 1 h before hypoxia-ischemia followed by aminoguanidine as in group A. Drugs and SF were administered for three consecutive days. On the tenth day, rat pups were decapitated and coronal sections at the level of dorsal hippocampal region of brains were evaluated. In the histopathologic examination; the mean infarcted area in group I+A was significantly lower than the control group (P<0.05). Although there was no statistically significant difference between treatment groups in terms of iNOS expression, the risk of iNOS expression was 7 times less for group I (CI: 1.6-30.8, P=0.01), 19.8 times less for group A (CI: 3.8-104, P=0.001) and 12.3 times less for group I+A (CI: 2.5-59, P=0.002) compared to group C. In conclusion, only indomethacin administration before hypoxic ischemia and followed by aminoguanidine was more effective to reduce infarct area, but we did not find any difference between treatment groups and control group for iNOS expression. So we suggest that this neuroprotection may not be related to depression of iNOS expression. PMID- 16258843 TI - Two-photon analysis of lead accumulation in rat cerebellar granule neurons. AB - Lead (Pb2+) is a common pollutant and potent central neurotoxin. We have studied its pathways of permeation by two-photon fluorescence microscopy in rat cerebellar granule neurons loaded with the fluorescent dye indo-1. Pb2+ binds indo-1 with high affinity acting as a quencher. Its permeation through the neuronal membrane was indicated by a decrease of the fluorescence emission, which occurred even in resting condition. In the presence of 20 microM Pb2+, uptake reached a plateau level (approximately 45% of initial fluorescence) in 4 min and was partially antagonized by 25 microM lanthanum. Subsequent addition of a membrane permeant ionophore caused a further (>70%) quenching of the dye, suggesting that previous saturation was due to inactivation of the transport system. Intracellular Pb2+ concentrations were evaluated from the fluorescence intensity and this estimate indicated that the concentration of free Pb2+ sufficient to inactivate the transport system is close to 50 pM. PMID- 16258842 TI - CYP46 T/C polymorphism is not associated with Alzheimer's dementia in a population from Hungary. AB - Multiple genetic and environmental factors regulate the susceptibility to Alzheimer's disease (AD). Recently, several independent studies have reported that a locus on chromosome 14q32.1, where a gene encoding a cholesterol degrading enzyme of the brain, called 24-hydroxylase (CYP46A1) is located, has been linked with AD. The single nucleotide polymorphism (T/C) in intron 2 of CYP46 gene has been found to confer the risk for AD. The water soluble 24(S)-hydroxysterol is the product of the CYP46A1, and elevated plasma and cerebrospinal fluid hydroxysterol concentrations have been found in AD, reflecting increased brain cholesterol turnover or cellular degradation, due to the neurodegenerative process. A case-control study was performed on 125 AD and 102 age- and gender matched control subjects (CNT) from Hungary, to test the association of CYP46 T/C and apolipoprotein E (ApoE) gene polymorphisms in AD. The frequency of the CYP46 C allele was similar (chi2=0.647, df=1, P=0.421, exact P=0.466, OR=0.845; 95% CI: 0.561-1.274) in both groups (CNT: 27%; 95% CI: 21.3-33.4; AD 30%; 95% CI: 25.0 36.3). The ApoE varepsilon4 allele was significantly over-represented (chi2=11.029, df=2, P=0.004) in the AD population (23.2%; 95% CI: 18.2-29.0) when compared with the CNT (11.3%; 95% CI: 7.4-16.6). The presence or absence of one or two CYP46C alleles together with the ApoE varepsilon4 allele did not increase the risk of AD (OR=3.492; 95% CI: 1.401-8.707; P<0.007 and OR=3.714; 95% CI: 1.549-8.908; P<0.003, respectively). Our results indicate that the intron 2 T/C polymorphism of CYP46 gene (neither alone, nor together with the varepsilon4 allele) does not increase the susceptibility to late-onset sporadic AD in the Hungarian population. PMID- 16258844 TI - Tau and GSK3beta dephosphorylations are required for regulating Pin1 phosphorylation. AB - Pin1 binds mitotically phosphorylated Thr231-Pro232 and Thr212-Pro213 sites on tau, and a Pin1 deficiency in mice leads to tau hyperphosphorylation. The aim of this study was to determine if the dephosphorylation or inhibition of tau and GSK3beta phosphorylation induces the Pin1 phosphorylation. To test this, human SK N-MC cells were stably transfected with a fusion gene containing neuron-specific enolase (NSE)-controlled APPsw gene(NSE/APPsw), to induce Abeta-42. The stable transfectants were then transiently transfected with NSE/Splice, lacking human tau (NSE/Splice), or NSE/hTau, containing human tau, into the cells. The NSE/Splice- and NSE/hTau-cells were then treated with lithium. We concluded that (i) there was more C99-beta APP accumulation than C83-betaAPP in APPsw tansfectant and thereby promoted Abeta-42 production in transfectants. (ii) the inhibition of tau and GSK3beta phosphorylations correlated with increase in Pin1 activation in NSE/hTau- cells. Thus, these observations suggest that Pin1 might have an inhibitive role in phosphorylating tau and GSK3beta for protecting against Alzheimer's disease. PMID- 16258845 TI - Demonstration of kynurenine aminotransferases I and II and characterization of kynurenic acid synthesis in oligodendrocyte cell line (OLN-93). AB - In the present study we demonstrate for the first time that both kynurenine aminotransferase (KAT) isoforms I and II are present in the permanent immature rat oligodendrocytes cell line (OLN-93). Moreover, we provide evidence that OLN 93 cells are able to synthesize kynurenic acid (KYNA) from exogenously added L kynurenine and we characterize its regulation by extrinsic factors. KYNA production in OLN-93 cells was depressed in the presence of aminotransferase inhibitor, aminooxyacetic acid and was not affected by depolarizing agents such as 50 mM K+ and 4-aminopyridine. Glutamate agonists, L-glutamate and D,L homocysteine significantly decreased KYNA production. Selective agonist of ionotropic glutamate receptors Amino-2,3-dihydro-5-methyl-3-oxo-4 isoxazolepropionic acid (AMPA) lowered KYNA production in OLN-93 cell line, whereas N-methyl-D-aspartate (NMDA) had no influence on KYNA production. Furthermore, KYNA synthesis in OLN-93 cells was decreased in a concentration dependent manner by amino acids transported by L-system, L-leucine, L-cysteine and L-tryptophan. The role of KYNA synthesis in oligodendrocytes needs further investigation. PMID- 16258846 TI - Ceramide induces non-apoptotic cell death in human glioma cells. AB - Ceramide causes either apoptosis or non-apoptotic cell death depending on model system and experimental conditions. The present study was undertaken to examine the effect of ceramide on cell viability and its molecular events leading to cell death in A172 human glioma cells. Ceramide induced cell death in a dose-dependent manner and the cell death was dependent on generation of reactive oxygen species and lipid peroxidation. TUNEL assay, Hoechst 33258 staining, and flow cytometric analysis did not show typical apoptotic morphological features. Ceramide caused phosphorylation of extracellular signal-regulated kinase (ERK) and p38, but the cell death was not affected by inhibitors of MAPK subfamilies. Ceramide caused ATP depletion without loss of mitochondrial membrane potential. Ceramide did not induce caspase activation and ceramide-induced cell death was also not altered by inhibitors of caspase activation. Transfection of dominant inhibitory mutant of IkappaBalpha (S32A/36A) and pretreatment of pyrrolidinedithiocarbamate, an inhibitor of NF-kappaB, enhanced ceramide-induced cell death. These results indicate that ceramide causes non-apoptotic, caspase-independent cell death by inducing reactive oxygen species generation in A172 human glioma cells. NF-kappaB is involved in the regulation of ceramide-induced cell death in human glioma cells. PMID- 16258847 TI - The effect of mild and severe hypoxia on rat cortical synaptosomes. AB - Brain ischemia results in neuronal injury and neurological disability. The present study examined the effect of mild (6% O2) and severe (2% O2) hypoxia on mitochondria of rat cortical synaptosomes. During mild and severe hypoxia, JO2 and ATP production significantly decreased and mitochondrial membranes depolarized. Synaptosomal calcium concentration increased slightly, albeit not significantly. After a 1 h re-oxygenation period, JO2, ATP production and mitochondrial membrane potential returned to control levels in synaptosomes incubated in 6% O2. In synaptosomes incubated in 2% O2, however, the ATP production was not restored after re-oxygenation and intrasynaptosomal Ca2+ significantly increased. The results indicate that both mild and severe hypoxia influence the physiology of synaptosomal mitochondria; the modifications are reversible after mild hypoxia and but partly irreversible after severe hypoxia. PMID- 16258848 TI - Glutamate decarboxylase: loss of N-terminal segment does not affect homodimerization and determination of the oxidation state of cysteine residues. AB - Glutamate decarboxylase (GAD) produces GABA, the main inhibitory neurotransmitter in adult mammalian brain. The physical characteristics of GAD were studied using mass spectrometry and partial protein digests. The N-termini of the two main isoforms, GAD65 and GAD67, were processed by removal of the initial methionine residues and acetylation of the penultimate alanines. Native recombinant GAD65 and GAD67 exist as homodimers that can be dissociated with non-reducing methods, indicating that homodimerization does not involve intermolecular disulfide bonds. Truncation of the N-terminal segment with trypsin digestion did not affect homodimerization but increased activity by decreasing the Km of GAD67 and increasing the Vmax of both isoforms. Of the 15 cysteines in GAD65, the six found in the N-terminal segment can form disulfide bonds and of the 13 cysteines in GAD67, cysteines 32 and 38 can form a disulfide bond. The in vitro formation of disulfide bonds in the N-termini, and the removal of the termini with relatively low amounts of trypsin, indicate that the N-terminal segments of GAD65 and GAD67 are exposed and flexible. The formation of a disulfide bridge between cysteines 30 and 45 of GAD65 suggests that alteration of normal redox conditions could affect GAD targeting. PMID- 16258849 TI - A study of glutathione S-transferase pi expression in central nervous system of subjects with amyotrophic lateral sclerosis using RNA extraction from formalin fixed, paraffin-embedded material. AB - The expression of glutathione S-transferase pi (GST pi), an enzyme responsible for inactivation of a large variety of toxic compounds was studied in spinal cord, motor and sensory brain cortex obtained from patients who died in the course of amyotrophic lateral sclerosis (ALS). The studies were performed on formalin-fixed, paraffin-embedded (FFPE) and freshly frozen tissues. The method of RNA isolation from FFPE was modified. A significant decrease of GST pi-mRNA expression was found in cervical spinal cord and motor brain cortex of ALS subjects comparing to analogue control tissues (P<0.01), as well as in motor cortex of ALS subjects comparing to their sensory cortex (P<0.05). In spinal cords the decrease in GST pi-mRNA expression was accompanied by a decrease of GST pi protein level. Results indicated lowered GST pi expression on both mRNA and protein levels in the regions of nervous system affected by ALS. The non-properly inactivated by GST toxic electrophiles and organic peroxides may thus contribute to motor neurons damage. PMID- 16258850 TI - Differentially expressed genes in transgenic mice carrying human mutant presenilin-2 (N141I): correlation of selenoprotein M with Alzheimer's disease. AB - Mutations in genes for Alzheimer's disease (AD) result in a modulating of gene expressions in the brains of patients with AD. The aim of this study was to identify genes whose expression is modulated due to the over-expression of human mutant presenilin-2 (N141I) (hPS2m) in transgenic mice, which has previously been produced by us. To test this, GeneFishing DEG101 technique was performed on large scale screen of mRNA from transgenic and non-transgenic brains. A total of 40 transcriptional products corresponding to cDNA were compared between two brains, and 17 showed a differential expression between the samples in all sets of experiments. However, all showed significant homology to known genes. Initially, a cloning corresponding to human selenoprotein M (hSelM) was chosen for investigation further because SelM induced by sodium selenite, a pro-oxidant, may have a functional role in catalyze the free radicals. We found that mouse SelM had significantly suppressed on its transcriptional products in transgenic brains. In parallel, suppression of endogenous was not observed in transgenic brains. Moreover, the levels of green fluorescence on hSelM fusion protein with EGFP were suppressed in the cells transfected with hPS2m, and its levels had actually increased by treatments of sodium selenite. Thus, the results indicate that SelM might play a suppressive or protective role in the pathology of patients with AD and it will be necessary to investigate further on functional roles of other up- and down-regulated gene in future. PMID- 16258851 TI - Effect of general anesthetics on amyloid precursor protein and mRNA levels in the rat brain. AB - The incidence of Alzheimer's disease is elevated after exposure to surgical interventions. Since amyloid precursor protein (APP) and its neurotoxic derivatives play key roles in the development of Alzheimer dementia, the role of general anesthesia is controversial in the development of cognitive decline. As such, the effect of anesthetics on APP protein and mRNA levels was assessed utilizing semiquantitative Western-immunoblot and reverse transcription polymerase chain reaction (RT-PCR) in brains of rats following intraperitoneal treatment with propofol and thiopental. The anesthetics did not change cortical APP protein and mRNA concentration considerably. These results indicate that both propofol and thiopental are considered to be relatively safe with respect to APP metabolism. PMID- 16258852 TI - Nicotine-induced sensitization in mice: changes in locomotor activity and mesencephalic gene expression. AB - It is believed that drug-induced behavioral sensitization is an important process in the development of substance dependence. In order to explore mechanisms of sensitization, a mouse model of nicotine-induced locomotor sensitization was established, and effects of the sensitization process on mesencepahlic gene expression were examined. A schedule, which included 3 weeks of intermittent nicotine exposure (0.5 mg/kg, s.c.) and 3 weeks of withdrawal, resulted in locomotor sensitization. Effects of sensitization on mesencephalic expression of approximately 14,000 genes were assessed using oligonucleotide microarrays. Signal intensity differences in samples obtained from repeated nicotine- and saline-exposed animals were analyzed with z-test after False Discovery Rate (FDR) multiple test correction. Genes related to GABA-A receptors and protein phosphatases were among 68 genes showing significantly different expression levels between the saline and the nicotine groups. We hypothesize that some of the gene expression changes in the mesencephalon are involved in pathways leading to nicotine-induced sensitization. Down-regulation of GABA-A receptors induced by repeated nicotine exposure may facilitate dopaminergic neuronal transmission and may contribute to increased locomotor activity. PMID- 16258853 TI - Agonistic properties of cannabidiol at 5-HT1a receptors. AB - Cannabidiol (CBD) is a major, biologically active, but psycho-inactive component of cannabis. In this cell culture-based report, CBD is shown to displace the agonist, [3H]8-OH-DPAT from the cloned human 5-HT1a receptor in a concentration dependent manner. In contrast, the major psychoactive component of cannabis, tetrahydrocannabinol (THC) does not displace agonist from the receptor in the same micromolar concentration range. In signal transduction studies, CBD acts as an agonist at the human 5-HT1a receptor as demonstrated in two related approaches. First, CBD increases [35S]GTPgammaS binding in this G protein coupled receptor system, as does the known agonist serotonin. Second, in this GPCR system, that is negatively coupled to cAMP production, both CBD and 5-HT decrease cAMP concentration at similar apparent levels of receptor occupancy, based upon displacement data. Preliminary comparative data is also presented from the cloned rat 5-HT2a receptor suggesting that CBD is active, but less so, relative to the human 5-HT1a receptor, in binding analyses. Overall, these studies demonstrate that CBD is a modest affinity agonist at the human 5-HT1a receptor. Additional work is required to compare CBD's potential at other serotonin receptors and in other species. Finally, the results indicate that cannabidiol may have interesting and useful potential beyond the realm of cannabinoid receptors. PMID- 16258854 TI - Effects of calmodulin and Ca2+ channel blockers on omega-conotoxin GVIA binding to crude membranes from alpha1B subunit (Cav2.2) expressed BHK cells and mice brain lacking the alpha1B subunits. AB - Characteristics for the specific binding of 125I-omega-CTX GVIA and 125I-omega CTX MVIIC to crude membranes from BHKN101 cells expressing the alpha1B subunits of Cav2.2 channels and from mice brain lacking the alpha1B subunits of Cav2.2 channels, particularly, the effects of CaM and various Ca2+ channel blockers on these specific bindings were investigated. Specific binding of 125I-omega-CTX GVIA to the crude membranes from BHKN101 cells was observed, but not from control BHK6 cells. omega-CTX GVIA, omega-CTX MVIIC and omega-CTX SVIB inhibited the specific binding of 125I-omega-CTX GVIA to crude membranes from BHKN101 cells, and the IC50 values for omega-CTXGVIA, omega-CTX MVIIC and omega-CTX SVIB were 0.07, 8.5 and 1.7 nM, respectively. However, omega-agatoxin IVA and calciseptine at concentrations of 10(-9)-10(-6) M did not inhibit specific binding. Specific binding was also about 80% inhibited by 20 microg protein/ml CaM. The amount of 125I-omega-CTX GVIA (30 pM) specifically bound to membranes from brain of knockout mice lacking alpha1B subunits of Cav2.2 channels was about 30% of that to the crude membranes from brain of wild-type. On the other hand, specific binding of 125I-omega-CTX MVIIC (200 pM) was observed on the crude membranes of both BHKN101 and control BHK6 cells. The specific binding of 125I-omega-CTX MVIIC (200 pM) was not inhibited by omega-CTX GVIA and omega-CTX SVIB, and also omega Aga IVA and calciseptine at concentrations of 10(-9)-10(-7) M, although specific binding was almost completely dose dependently inhibited by non-radiolabeled omega-CTX MVIIC (IC50 value was about 0.1 nM). 20 microg protein/ml CaM did not inhibit specific binding. Therefore, these results suggest that BHKN101 cells have a typical Cav2.2 channels which are also inhibited by CaM and have not specific binding sites for omega-CTX MVIIC, although omega-CTX MVIIC is a blocker for both Cav2.1 (alpha1A; P/Q-type) and Cav2.2 channels. PMID- 16258855 TI - Locally administered low nicotine-induced neurotransmitter changes in areas of cognitive function. AB - The present study examined the effect of a low-dose of nicotine; below that one expects to be achieved from a single cigarette, on brain regional heterogeneity and sensitivity of catecholaminergic responses. 1 microM nicotine was infused into six brain areas via a microdialysis probe: the dorsal and ventral hippocampus, the medial temporal and prefrontal cortex, the basolateral amygdala, and the ventral tegmental area (VTA). The nicotine concentration in the brain tissue near the probe site was approximately 0.1 microM. Nicotine-induced increases and decreases could be noted in dopamine (DA), norepinephrine (NE), and serotonin (5HT) levels. In particular, DA and 5HT decreased in both hippocampal areas, while NE increased in the dorsal and decreased in the ventral hippocampus. In the cortical areas, DA and NE increased and 5HT was not significantly altered. In the amygdala all three neurotransmitters increased and in the VTA, all three decreased. Many of the nicotine-induced changes in neurotransmitter concentrations were reversed in the presence of atropine. Where nicotine induced decreases in DA and 5HT in the VTA, increases were observed in the presence of atropine. A similar reversal was seen with NE in the VTA and ventral hippocampus. In contrast, the increases in DA observed in the cortex and amygdala and the increases in NE observed in the cortex, amygdala and dorsal hippocampus were inhibited by the presence of atropine. 5HT was also significantly decreased in the amygdala and both cortical areas in the presence of atropine, where nicotine alone had no significant effect. We conclude, that at low doses, nicotine significantly alters the release of DA, NE, and 5HT--in some areas increasing, in others decreasing endogenous neurotransmitter levels. This data, in conjunction with previous experiments, indicates that the effects of nicotine are regionally heterogeneous and arise from both direct and indirect actions on various receptors and neurotransmitter systems and nicotine's effects at low doses differ from that at higher doses. The changes in effects in the presence of atropine suggest that muscarinic acetylcholine receptors play a major role in nicotine's actions on neurotransmitter systems. PMID- 16258857 TI - Evidence of recent progress in clinical practice and research in infectious disease without official measures or meddling -- an example for Germany. PMID- 16258856 TI - Citicoline inhibits MAP kinase signalling pathways after focal cerebral ischaemia. AB - The link between membrane phospholipids and different intracellular signal transduction pathways affected by cerebral ischaemia is unclear. CDP-choline, a major neuronal membrane lipid precursor and its intracellular target proteins and transcription factors were studied to further understand its role in ischaemic stroke. Cerebral ischaemia was produced by distal, permanent occlusion of the middle cerebral artery (MCAO) in the rat. Animals receiving 500 mg/kg of CDP choline in 0.5 ml of 0.9% saline, intraperitoneally, 24 h and 1 h before MCAO and 23 h after MCAO demonstrated a notable reduction in the phosphorylation of MAP kinase family members, ERK1/2 and MEK1/2, as well as Elk-1 transcription factor, compared with control animals treated with 0.5 ml of 0.9% saline. Immunohistochemistry showed a particular reduction in immunoreactivity in glia. The effects of CDP-choline on intracellular mechanisms of signal transduction, suggests that this molecule may play a key role in recovery after ischaemic stroke. PMID- 16258859 TI - Dutch guideline for preventing nosocomial transmission of highly resistant microorganisms (HRMO). AB - Hospitals are faced with the increasingly rapid emergence and dissemination of antimicrobial-resistant microorganisms. US and European guidelines on the prevention of antimicrobial resistance in hospitals were, until recently, mainly directed at methicillin-resistant Staphylococcus aureus (MRSA). In 2004, the Dutch Working Party on Infection Prevention issued a guideline on the prevention of nosocomial transmission of highly resistant microorganisms (HRMO), in order to fulfill the growing need for additional guidance on the control of other pathogens with acquired resistance and the potential to spread within hospitals (such as glycopeptide-resistant Enterococcus faecium, penicillin-resistant Streptococcus pneumoniae, extendedspectrum beta-lactamase producing Enterobacteriaceae, and other (multi)drug-resistant gram-negatives). In addition to providing criteria for defining HRMO, the Dutch guideline provides recommendations on isolation of patients, active surveillance, and contact tracing. The guideline will enable the comparison of HRMO rates between hospitals, and may be used to evaluate the efficacy of programs to control antibiotic use and/or nosocomial transmission of resistant pathogens. The eventual success of nationwide implementation of this guideline remains to be established in the coming years. PMID- 16258860 TI - Low rate of clinical consequences derived from results of blood cultures obtained in an internal medicine emergency department. AB - BACKGROUND: Blood cultures detect bacteremia in individual patients and help define local pathogen and resistance spectra. At the same time, the benefits of blood culture results in the management of individual patients -- and therefore their cost-effectiveness -- are disputed. PATIENTS AND METHODS: During 1 calendar year, we conducted a prospective study of emergency department admissions with blood culture draws and at least a 3-day hospitalization afterwards. We prospectively surveyed treating physicians on usefulness of blood culture results for patient management. RESULTS: 428 diagnostic episodes (emergency visits) involving 390 patients occurred during the study period from 10/2002 to 10/2003. The analysis included 188/428 (44%) episodes with blood culture draws performed according to the predefined clinical standard where patients were hospitalized with sufficient duration. Absence of therapeutic consequences in response to blood culture results was reported for 138/142 (97%) of episodes with negative blood culture results, for 16/21 (76%) with blood culture results positive only for skin flora, and for 14/25 (56%) of episodes with blood cultures positive for obligate pathogens. Treating physicians regarded the blood culture results necessary for clarifying the etiology in 34/188 (18%) episodes, and rated blood culture results necessary for their therapeutic decisions in 29/188 (15%) episodes. CONCLUSION: Negative blood culture results rarely changed the management of medical inpatients. Our study suggests that in settings with broad spectrum empirical antibiotic therapy positive blood culture results for obligate pathogens trigger adjustment of the antibiotic therapy in only about half of instances. Many blood cultures drawn in the emergency department where considered unnecessary by ward physicians. Guidelines for preventing unnecessary blood culture draws are warranted in order to increase the rate of their meaningful clinical consequences for medical inpatients initially treated with broad spectrum empirical antibiotics. PMID- 16258861 TI - Antibiotic-resistant bloodstream infections in hospitalized patients: specific risk factors in a high-risk population? AB - BACKGROUND: The aim of this study was to explore characteristics that are associated with bloodstream infections due to specific multiresistant microorganisms (methicillinresitant Staphylococcus aureus, MRSA; vancomycin resistant enterococci, VRE; third-generation cephalosporin-resistant Enterobacteriaceae) or Candida spp. in hospitalized patients. PATIENTS AND METHODS: All patients who experienced a bloodstream infection with one of the aforementioned pathogens between September 1999 and October 2001 were included into a statistical analysis of independent risk factors. The possible impact of previous antibiotic and antifungal therapies was evaluated. RESULTS: Of the study population, 22% had two or more episodes with different pathogens. In the 314 patients with a single bloodstream infection MRSA was isolated in 189 patients, VRE in 31, Enterobacteriaceae in 13, and Candida spp. in 80 patients. Crude mortality was high in the study population (overall 40%) and varied between 33% (MRSA bacteremia only) and 58% (VRE bacteremia only). Patients who yielded more than one of the pathogens under surveillance had crude mortalities ranging from 41% to 83% (all four pathogens). In this group of high-risk patients, the following factors were independently associated with the individual pathogen: prior chemotherapy (OR 4.88 CI(95) 1.50-15.87) and bronchoscopy (OR 3.17 CI(95) 1.05-9.52) for VRE patients; burns (OR 4.50 CI(95) 0.90-22.73), presence of a tracheostomy (OR 4.22 CI(95) 1.15-15.38) and acute dialysis (OR 3.62 CI(95) 0.99 13.16) for patients with Enterobacteriaceae; and an underlying malignant disease (OR 1.98 CI95 0.99-3.97), performance of a bowel endoscopy (OR 2.80 CI(95) 1.27 6.13) and presence of a central venous catheter (CVC) (OR 12.34 CI(95) 1.63 90.91) for patients with candidemia. CONCLUSION: Patients with bacteremia due to VRE, Enterobacteriaceae or Candida spp. had more severe risk factors associated with the respective pathogen than patients with MRSA bacteremia. PMID- 16258862 TI - Risk factors for colonization with third-generation cephalosporin-resistant enterobacteriaceae. AB - BACKGROUND AND METHOD: Colonization and infections caused by Enterobacteriaceae resistant to third-generation cephalosporins (CRE) have been observed with increasing frequency in intensive care unit (ICU) patients. In contrast to outbreak investigations, information about risk factors for colonization in an endemic situation are rare. We studied risk factors for colonization with CRE in a case control study including 1,706 patients, admitted to any of the 15 ICUs of Heidelberg University Hospitals. RESULTS: 163 patients carried CRE with Enterobacter spp. representing the predominant species. Independent risk factors for CRE carriage in the multivariate logistic regression analysis were an age of under 2.5 years (OR 4.034), an indwelling central venous catheter (CVC) for more than 3 days (OR 2.640), treatment with second- or third generation cephalosporin for longer than 3 days (OR 2.260) and any antibiotic therapy before admission to the ICU. CONCLUSION: Apart from the well-recognized risk factor previous antibiotic treatment, the risk factors age and presence of a CVC might suggest that bacterial overgrowth of the gut either due to an increased susceptibility in younger age or as a consequence of parenteral nutrition is a relevant mechanism for acquiring carriage of CRE in a non-outbreak situation. PMID- 16258864 TI - Prevalence of Clostridium difficile toxins A and B and Clostridium perfringens enterotoxin A in stool samples of patients with antibiotic-associated diarrhea. AB - BACKGROUND: Antibiotic-associated diarrhea (AAD) is a major nosocomial as well as a community health problem. Clostridium difficile toxins (CDT) can be detected in only 10-25% of patients with AAD. The role of Clostridium perfringens enterotoxin A (CPEnt) as a cause of AAD remains to be elucidated. We, therefore, prospectively investigated the prevalences of both CPEnt and CDT in stool samples of patients with AAD. MATERIALS AND METHODS: A total of 693 stool samples consecutively submitted to our department from patients with AAD were screened for CDT and CPEnt using commercially available enzyme-linked immunoassays (ELISA). C. difficile and C. perfringens were detected by standard culture techniques. In addition, samples being CPEnt positive and/ or harboring C. perfringens were screened for the CPEnt gene by duplex PCR. RESULTS: CDT was detected in 79 (11.4%) of 693 stool samples. Of these, 49 (62.0%) harbored C. difficile. In one (0.14%) of 693 samples, CPEnt could be detected by ELISA. This respective CPEnt-positive stool sample also harbored C. perfringens. 147 (21.2%) of all stool samples were culture positive for C. perfringens. We did not detect samples positive for both CPEnt and CDT. In five (3.4%) of 147 C. perfringens isolates, the CPEnt gene could be detected by duplex PCR. PCR was positive in two (40%) of the five stool samples harboring CPEnt gene-positive C. perfringens isolates. CONCLUSION: The present prospective study revealed a prevalence of CDT of 11.4%, whereas the prevalence of CPEnt was less than 1%. Routine screening of stool samples for CPEnt does not appear to be justified in patients with AAD. PMID- 16258863 TI - Antibiotic use in non-university regional acute care general hospitals in southwestern Germany, 2001-2002. AB - BACKGROUND: A previous study from Germany showed high antibiotic use in university hospitals, particularly in intensive care units (ICU) and hematology oncology services, but there has been no information about recent antibiotic use in non-university hospitals. In the present study, we collected data from 40 non university regional general hospitals located in the southwestern part of the country, and analyzed use density in the medical and surgical services of these hospitals. MATERIALS AND METHODS: Hospital pharmacy records for the calendar years 2001 and 2002 were evaluated. The number of defined daily doses (DDD, definition according to the WHO/ATC 2001 index) and prescribed daily doses (PDD) per 100 patient days (DDD/100 or PDD/100, respectively) were calculated to compare antibiotic use densities in medical and surgical services. Data for surgery included various subspecialties and gynecology. RESULTS: Antibiotic use in the participating hospitals increased minimally between 2001 and 2002 both in medicine as well as in surgery. Use density in internal medicine (ICU areas excluded) in the year 2002 ranged between 13.5 and 93.7 DDD/100 with a weighted mean of 49.9 DDD/100 (corresponding to 28.6 PDD/100, respectively). Values for surgery were lower with a weighted mean of 43.4 DDD/100 (corresponding to 26.1 PDD/100, range, 10 to 65.4 DDD/100), respectively. Hospital size was not a strong predictor of use density, while large differences were observed between intensive care areas and normal wards. Mean use densities in intensive care areas in 2002 were 105.6 DDD/100 (or 49.7 PDD/100) in medical intensive care units, 116.9 DDD/100 (or 61.2 PDD/100) in surgical intensive care units, and 112.7 DDD/100 (or 66.7 PDD/100) in mixed, interdisciplinary intensive care units. Betalactams made up > 50% of all PDDs, while fluoroquinolones were the second most frequently prescribed drugs (15% of all PDDs). Fluoroquinolones were usually given orally. Overall glycopeptide and aminoglycoside use was < 1 PDD/100. CONCLUSION: This recent data from a large regional nonuniversity acute care hospital sample confirms that hospital antibiotic use density largely depends on patient care areas and less on hospital size. Surprisingly low use was observed for glycopeptides and aminoglycosides. The data may be useful as a benchmark for further pharmaco-epidemiologic evaluation and focused drug use control interventions. PMID- 16258865 TI - Incidence and prognosis of CMV disease in HIV-infected patients before and after introduction of combination antiretroviral therapy. AB - BACKGROUND: Highly active antiretroviral therapy (HAART) has improved the prognosis of HIV-infected patients. We studied the changes in the incidence and prognosis of cytomegalovirus (CMV) disease preceding and during the first few years of HAART in a clinic cohort. PATIENTS AND METHODS: All patients with CMV disease diagnosed between 1993 and 1999 from a clinic cohort in Cologne, Germany, were included. The patients were followed until death or until December 31, 2001. The time period from 1993-1996 was classified as pre-HAART, the period from 1997 1999 as the HAART era. Survival was analyzed with a Cox-proportional hazard model. RESULTS: From a total of 1,279 HIV-infected patients, 127 patients with CMV disease were enrolled. The incidence of CMV disease declined rapidly and significantly from 7.34 cases per 100 patient years (py) in the pre-HAART era to 0.75 cases per 100 py in the HAART era. The median survival time in the pre-HAART era was 9.5 months; the median survival was not yet reached at 4 years of follow up in the HAART era. The only risk factors influencing survival were CD4-cell count and antiretroviral therapy before and after diagnosis of CMV disease. Treatment naive patients had a better prognosis than pretreated patients and patients treated with triple combination therapy survived longer than patients with other treatment modalities. CONCLUSION: A rapid decline in the incidence of new CMV manifestations and a better prognosis of patients with CMV disease, especially if they were treatment naive and treated with triple combination therapy, were observed in the HAART era. PMID- 16258866 TI - Prospective surveillance of nosocomial infections in a Swiss NICU: low risk of pneumonia on nasal continuous positive airway pressure? AB - BACKGROUND: This study assessed the rate of invasive nosocomial infections in very low birth weight (VLBW) 1,500 g, who had received a central venous or umbilical catheter, or assisted ventilation. Nosocomial infections (sepsis, pneumonia, necrotizing enterocolitis [NEC]) were defined according to Centers for Disease Control (CDC) recommendations with slight modifications and their rates measured longitudinally. RESULTS: Among VLBW neonates, 16 nosocomial infections for an overall infection rate of 6 per 1,000 patient days were found. Infants with infection were of lower birth weight, a greater proportion was male, received lipid infusions, and on average had a higher severity of illness (CRIB) score. Interestingly, the ventilator-associated pneumonia (VAP) rate (12.5/1,000 ventilator days) seemed significantly higher than the pneumonia rate during nasal continuous positive airway pressure (NCPAP) treatment (1.8/1,000 NCPAP days; p = 0.04). The sepsis rate associated with peripheral catheters almost equaled the central line-associated rate, although numbers for both device-related infections were small. CONCLUSION: Further studies are needed to confirm the observation that the NCPAP-associated pneumonia rate might be lower than the VAP rate in VLBW infants, as well as to confirm the second observation that the sepsis rates on peripheral catheters compared to central venous catheters might be almost equal in VLBW infants. Reducing the exposure to ventilation via endotracheal tube, but not using peripheral as opposed to central catheters, might reduce the incidence of device-associated infection in this patient population. PMID- 16258867 TI - Pseudomonas aeruginosa strains from nosocomial pneumonia are more serum resistant than P. aeruginosa strains from noninfectious respiratory colonization processes. AB - BACKGROUND: Serum resistance is regarded as a major virulence factor of bacteria and is thought to be mediated by O side chains of the lipopolysaccharides (LPS). We investigated the serum-resistance properties and O serogroups of Pseudomonas aeruginosa strains isolated from intensive careunit (ICU) patients with pneumonia and from the respiratory tract of ICU patients without respiratory tract infections. MATERIALS AND METHODS: 171 P. aeruginosa strains were consecutively isolated from bronchoalveolar lavage fluid or transtracheal aspirates of ICU patients with monobacterial nosocomial pneumonia and 49 strains were isolated from the respiratory tract of ICU patients without respiratory tract infections. All strains were O serogrouped using Oantigen- specific sera for 14 O serogroups and tested for their sensitivity to the serum's bactericidal effect. RESULTS: Using two different analyses, the frequency of serum-sensitive isolates was significantly lower in strains from patients with pneumonia (56.1%; n = 96/171 and 22.8%, n = 39/171, respectively) than in strains from asymptomatically colonized patients (73.46%; 36/49 and 38.8%, n = 19/49, respectively) (p = 0.03; OR = 2.163; 95% CI = 1.072-4.368 and p = 0.0289; OR = 2.144; 95% CI = 1.089 4.368, respectively). O serogrouping revealed higher frequency of the serogroups A (11.9% and 16.3%, respectively), B (14.3% and 21%), E (26.5% and 24.6%), and I (28.6% and 28%) in both strain collections. The frequency of serum-sensitive strains (13/28 and 3/45, respectively) was significantly lower among strains expressing the A and B serogroups, than for all other serogroups (p < 0.05). CONCLUSION: Strains isolated from patients with pneumonia and strains possessing O-A or O-B serogroups appear to have greater pathogenic potential by virtue of their ability to resist serum-mediated killing. The linkage, however, between the O serogroups, serum resistance, and a strain's virulence remains unclear at this stage. PMID- 16258868 TI - Silencing of Bak ameliorates apoptosis of human proximal tubular epithelial cells by Escherichia coli-derived Shiga toxin 2. AB - BACKGROUND: Escherichia coli-derived Shiga toxin (Stx), the cause of the enteropathic hemolytic uremic syndrome, is a potent inducer of apoptotic cell death. The present study was performed to examine the hypothesis that Stx initiates apoptosis by activating the mitochondrial pathway involving mitochondrial-associated, pro-apoptotic Bcl-2 family proteins Bax and Bak. MATERIALS AND METHODS: To determine if Stx2-mediated apoptosis is dependent on Bax or Bak, a gene-silencing approach was employed using sequence-specific small interfering (si)RNA duplexes. Silencing of Bax and Bak protein expression in human renal proximal tubular epithelial (HK-2) cells and its effect on Shiga toxicity was assessed by immunofluorescence microscopy and Western blotting. RESULTS: Transfection of HK-2 cells, shown to be exquisitely sensitive to Stx, with siRNA duplexes successfully diminished Bak, but not Bax protein expression. In order to determine if silencing of pro-apoptotic gene expression affects Stx induced apoptosis, HK-2 cells were transfected with Bak-specific or control siRNA, exposed to lethal concentrations of Stx2 and assessed for cleavage of poly(ADPribose) polymerase-1 (PARP) as a marker of apoptosis, using Western blot technology. We observed that siRNA-induced reduction of Bak expression levels correlated with decreased PARP cleavage. CONCLUSION: Results suggest that Stx induced cell death involves pro-apoptotic Bak and that silencing of Bak gene expression affords partial protection against Stx-mediated apoptosis. PMID- 16258869 TI - Inducible metronidazole resistance in nim-positive and nim-negative bacteroides fragilis group strains after several passages metronidazole containing columbia agar plates. AB - BACKGROUND: Recent data show an emergence of resistance in the Bacteroides fragilis group against several antimicrobial agents and inducible resistance against metronidazole in nim-positive strains. The aim of the present study was to investigate inducible metronidazole resistance in nim-positive as well as in nim-negative B. fragilis group strains. MATERIALS AND METHODS: Of 18 B. fragilis strains (including four nim-positive reference strains and one ATCC strain), two Bacteroides ovatus strains, and one Bacteroides thetaiotaomicron DSM strain minimum inhibitory concentration (MIC) values for metronidazole were determined by Etest and analyzed for nim genes (nimA to -G) by PCR. For this purpose bacterial suspensions were incubated on supplemented Columbia agar plates containing metronidazole at twice the MIC value of the specific strain and incubated under anaerobic conditions for 48 hours. After incubation, growing bacteria were harvested and thereafter incubated at four times the original MIC. This procedure was repeated with increasing antibiotic concentrations. The resulting MIC values were confirmed by Etest. RESULTS: The MIC values for metronidazole of the four nim-positive reference strains ranged from 3 to 8 mg/l. The B. fragilis ATCC 25285 strain and the B. thetaiotaomicron DSM 2255 strain were nim negative with MIC values of 0.19 mg/l and 0.75 mg/l, respectively. Three clinical isolates of B. fragilis strains showed MIC values of > 256 mg/l. In all three strains, nim genes were detected by PCR. The other clinical isolates were nim negative. In these strains, MIC values ranged from 0.19 to 0.75 mg/l. After several passages on metronidazole containing agar, all B. fragilis group strains exhibited MIC values of > 256 mg/l determined by Etest. CONCLUSION: Metronidazole resistance can be selected not only in nim-positive strains but also in nim negative strains, suggesting that mechanisms other than nim genes are involved. These findings and the emerging resistance of the B. fragilis group against several antimicrobial agents underscore the importance of susceptibility testing of anaerobes even in routine laboratories. PMID- 16258870 TI - Analysis of the specific immune response against capsular polysaccharides of two patients with systemic enterococcal infections. AB - Systemic enterococcal infections often lead to life-threatening disease. By analyzing the immune response of two patients with systemic enterococcal infections against enterococcal polysaccharide antigens, we found that both patients had antibodies against all four of the capsular serotypes identified to date. Antibody concentrations against the causative capsular serotype were in the same range as antibodies against the other three capsular protoserotypes. Interestingly, we noted a difference between the two patients with respect to opsonic activity in the killing assay: one patient showed better killing of all four capsular prototypes than the other. However, killing against the infecting serotype was not increased in comparison to killing of the other serotypes in the two patients. This finding supports previously published data that most healthy humans possess preexisting, naturally acquired, anti-enterococcal antibodies. We conclude, therefore, that systemic infection with enterococci does not lead to higher antibody concentrations or better opsonic killing against the causative capsular serotype. PMID- 16258871 TI - In vitro assessment of the host response against Enterococcus faecalis used in probiotic preparations. AB - Along with other lactic acid bacteria, enterococci are used in food products and as health promoting agents. The safety of these products must be ensured, because they contain potentially pathogenic microorganisms. Here we present an in vitro opsonophagocytic assay that closely mimics the protective human immune response to Enterococcus faecalis and Enterococcus faecium. A collection of closely related E. faecalis isolates used as probiotics showed different susceptibilities to opsonic killing, suggesting that some of these isolates possess a capsule while other do not. This information may be helpful in assessing the safety of a given bacterial isolate used and could detect likely enterococcal candidates for probiotic preparations. PMID- 16258872 TI - Interleukin-8 serum levels at fever onset in patients with neutropenia predict early medical complications. AB - BACKGROUND: Previous studies have shown that interleukin-8 serum levels in febrile neutropenic patients are significantly higher in patients with gram negative bacteremia than in patients with other causes of fever and may indicate unfavorable outcomes. We assessed the value of interleukin-8 serum levels at fever onset to predict clinical complications in order to confirm these earlier findings. PATIENTS AND METHODS: In a prospective observational study of adult patients receiving cancer chemotherapy, serum samples obtained at the onset of 147 febrile neutropenic episodes were measured by an immunoluminescence assay. RESULTS: Complicated courses of fever including severe sepsis or septic shock, respiratory insufficiency or death were observed in 13 episodes (9%); in six episodes complications had developed within 1 week after fever onset and five of them were associated with bloodstream infections. At an interleukin-8 cutoff level of 1,000 pg/ml, these early complications were predicted with a sensitivity of 83%, a specificity of 97%, a positive predictive value of 50%, and a negative predictive value of 99%, respectively. CONCLUSION: Interleukin-8 levels at fever onset may be used for the prediction of early medical complications associated with bacteremia and can help identify patients who might benefit from intensive care admission. PMID- 16258873 TI - Fatal outcome of a hyperinfection syndrome despite successful eradication of Strongyloides with subcutaneous ivermectin. AB - We report the case of a 77-year-old man who developed a Strongyloides hyperinfection syndrome following immunosuppressive therapy more than 60 years after he moved away from an area endemic for Strongyloides stercoralis. Successful eradication of the nematode was achieved with an off label subcutaneous formulation of ivermectin. However, the patient subsequently died from acute respiratory distress syndrome (ARDS). Despite a high wormload in the stool and sputum of the patient and delayed infection control measures in the hospital, testing of the medical staff revealed a very low risk of Strongyloides transmission among healthcare workers. PMID- 16258874 TI - Aortitis diagnosed by F-18-fluorodeoxyglucose positron emission tomography in a patient with syphilis and HIV coinfection. AB - The most common manifestation of cardiovascular syphilis, a rare diagnosis since the introduction of penicillin, is aortitis of the ascending aorta. Since the majority of patients with uncomplicated aortitis are asymptomatic, early diagnosis is difficult. We report the case of an HIV-positive patient with asymptomatic syphilitic aortitis that was incidentally diagnosed with F-18 fluorodeoxyglucose (FDG) positron emission tomography (PET). We conclude that FDG PET could become a promising new imaging technique for both diagnosis and follow up of patients with syphilitic aortitis. PMID- 16258875 TI - Peritonitis due to Neisseria mucosa in an adolescent receiving peritoneal dialysis. AB - Neisseria mucosa is part of the normal nasopharyngeal flora and rarely pathogenic in humans. Reports of serious infections associated with this pathogen are very unusual. A 17-year-old boy with end-stage renal disease due to IgA nephropathy presented with acute, spontaneous, symptomatic peritoneal dialysis-associated peritonitis without reported break in sterility or PD catheter exit site infection. beta-lactamase-negative N. mucosa was isolated from the dialysate effluent. Intraperitoneal antibiotic treatment with cephalothin/gentamicin for 5 days and subsequent ceftriaxone led to complete resolution of the infection. This case demonstrates that "non-pathogenic" Neisseria species can cause clinically severe peritonitis with high intraperitoneal neutrophil counts, elevated C reactive protein levels in the peritoneal effluent (in the presented case, 27,600/mul and 3.6 mg/l, respectively) and impaired peritoneal membrane transport function. To our knowledge, this is the first case of N. mucosa peritonitis complicating chronic peritoneal dialysis in an adolescent patient. PMID- 16258876 TI - Landouzy septicemia (sepsis tuberculosa acutissima) due to Mycobacterium microti in an immunocompetent man. AB - Even in developed countries, tuberculosis still contributes significantly to morbidity and mortality. The most frequent causative agent is Mycobacterium tuberculosis, while infections due to other mycobacterial species are usually associated with immunocompromised patients. In the following, we describe the case of a previously healthy man who underwent laparotomy for suspected adrenal carcinoma. Peritoneal "cancerous nodules" turned out to be tuberculous granulomas. After surgery the patient developed a protracted septic shock and died 6 days after surgery. Isolation and identification of the causative agent yielded Mycobacterium microti, an uncommon species of the M. tuberculosis complex. No other pathogen could be isolated during the clinical course, which finally led to the diagnosis of Landouzy septicemia (sepsis tuberculosa acutissima). PMID- 16258878 TI - What's new in HIV/AIDS? Neutralizing HIV antibodies: do they really protect? AB - Neutralizing antibodies work as a second line of defence. They are detected more or less in nearly every HIV-1-infected individual. 2G12, 2F5 and 4E10 represent antibodies with broadly neutralizing activity made from B cells of HIV- 1 infected humans. Unfortunately these antibodies are extremely rare and all attempts to elicit them via vaccine immunogens have failed. The discovery of autoreactive features of these antibodies could now explain why. Additionally, new results show the delayed viral rebound under antibody treatment in some HIV-1 infected individuals. PMID- 16258877 TI - Breakthrough invasive infection due to Debaryomyces hansenii (teleomorph Candida famata) and Scopulariopsis brevicaulis in a stem cell transplant patient receiving liposomal amphotericin B and caspofungin for suspected aspergillosis. AB - An allogeneic stem cell transplant recipient developed pulmonary infiltrates and Aspergillus antigenemia during prophylactic low-dose liposomal amphotericin B. No response to therapy was observed after increasing the dose of liposomal amphotericin B and addition of caspofungin, and breakthrough candidemia developed. Therapy switch to voriconazole did not prevent the development of lethal septic shock. Shortly before death, Scopulariopsis brevicaulis was cultured from bronchial secretions, and positive blood cultures demonstrated persistent candidemia due to Debaryomyces hansenii, teleomorph of Candida famata. PMID- 16258879 TI - What's New in HIV/AIDS? Chemokine receptor antagonists: a new era of HIV therapy? AB - Chemokine receptors are essential for human immunodeficiency virus (HIV) cell entry. CXCR4 and CCR5 are the two most relevant receptors and by inhibition of each of them a delayed onset of disease could be achieved. As both receptors are used at different stages of disease due to the domination of different HIV strains, a dual blockage of CXCR4 and CCR5 could be highly valuable for inhibiting viral transmission and replication. PMID- 16258880 TI - Can micronucleus technique predict the risk of lung cancer in smokers? AB - Smoking is the main etiological factor in the carcinogenesis process of lung cancer. But genetically defined factors such as increased levels of oxidase enzymes or chromosome aberrations have been shown to correlate with the higher possibility of contracting lung cancer among smokers. In this study, chromosome aberrations measured by micronucleus (MN) technique following in vitro irradiation were investigated in peripheral blood lymphocytes of long term smokers with or without lung cancer. Our aim is to establish the role of MN scores in identifying the individuals who might develop cancer among smokers. Twelve lung cancer patients and appropriately matching 10 healthy controls were evaluated. Spontaneous and radiation induced MN frequencies were evaluated in the two groups. An increase in the amounts of MN after 3 Gy irradiation was observed in the patient and control group when compared to spontaneous frequencies. Absolute MN frequencies as a determinant of radiosensitivity were calculated by subtraction of spontaneous aberration frequencies from the frequencies that were obtained following 3 Gy of irradiation. Absolute MN frequency range was between 0.0116 and 0.3883 with the average value of 0.1114 +/- 0.0390 (SE) for the lung cancer patients, and was between 0.0216 and 0.2291 with the average value of 0.1410 +/- 0.0234 (SE) for the controls. When the comparison was made between the absolute MN frequencies of both groups, there was no difference (p=0.159) between the two groups. In our study, it can be concluded that radiation induced MN scores in peripheral blood lymphocytes of long term smokers do not predict the risk of lung cancer. PMID- 16258882 TI - Diagnostic yield of closed pleural brushing. AB - The aim of this study was to assess the diagnostic yield of closed pleural brushing (CPBR) in the diagnosis of malignant pleural effusion. Twenty-one adult patients (20 men and 1 woman); aged 62.9 +/- 8.6 were participated to this prospective study. Thoracentesis, CPBR and closed pleural biopsy (CPB) following the brushing were applied to every patient. While CPBR provided diagnosis in 12 (57.1%) of 21 cases, in 3 of these 12 cases, pleural fluid cytology (PFC) and CPB were negative. The sensitivities of PFC, CPBR and CPB in the diagnosis of malignant effusions were 33%, 57% and 52%, respectively. When three procedures were used in combination, the sensitivity increased to 67%. When CPBR is performed in addition to PFC and CPB, the yield of the diagnosis increased 14% additionally. There was no mortality due to these interventions. Complications were chest pain in 3 (14.2%) cases, hypotension in 2 (9.5%) cases, cough in 1 (4.8%) case, pneumothorax in 1 (4.8%) case, and hemothorax in 1 (4.8%) case. In conclusion, CPBR as a safe, simple and well tolerated procedure provides high diagnostic yield in diagnosis of patients with malignant pleural effusion. PMID- 16258881 TI - The detection of quantitative serum p53 protein in lung cancer. AB - p53 protein, which accumulates intracellularly in over half of all human tumors, has been reported to be variably present in the sera of patients with various malignancies. In this study, it was aimed to detect p53 protein in the sera of lung cancer patients, and to verify its value as a marker of p53 alterations in lung cancer. A pantropic quantitative ELISA technique was used to detect serum p53 protein of 94 newly diagnosed patients with lung cancer. Serum samples were collected on admission before any treatment. There was no detectable serum p53 protein in the control group including 34 healthy volunteers. Serum p53 protein was present in only 3 (3.2%) of 94 patients. In nonsmall cell lung cancer (NSCLC) group, serum p53 protein had been detected in 2 (2.8%) of 72 patients, and it was detectable in 1 (4.5%) of 22 patients in SCLC group. Serum levels of p53 protein ranged from 1 U/mL to 31.25 U/mL in positive samples. Patients who had p53 protein in their serum samples, were at late stage and had poor prognosis. In conclusion; prognostic value of detectable serum p53 protein levels could not be define, because of the small number of p53 positive patients. The use of quantitative serum p53 protein analysis with ELISA is of very limited value as a marker in evaluating p53 changes in lung cancer patients, despite the fact that is an easy technique to perform. PMID- 16258883 TI - [Investigating the anti-inflammatory effect of dexamethasone in an asthma mouse model]. AB - We performed an asthma mice model in this study and aimed to investigate the levels of mediators in bronchoalveolar lavage fluid (BALF), and lung tissue, and the pathological changes response to the steroid treatment. BALB/c mice divided into three groups. PBS was applied to group 1 (control group). Asthma model was performed by exposing to ovalbumin in group 2 and 3. DEX was injected to group 3. After the last DEX dose all of the mice were killed by cervical dislocation. The samples of BALF and lung tissue were obtained. IL-4 and IL-5 levels of all samples were measured and inflammatory cells were counted in BALF. Evident eosinophilia was determined in BALF of group 2. Eosinophil numbers were lower in group 3 when compared with group 2 and this was statistically significant (p< 0.001). Inflammatory cell infiltration, eodema and hyperemia observed around the walls of bronchus and bronchiols in group 2. The lungs of group 3 had normal histological appearance. Both two cytokin levels of lung tissue were higher in group 2 than group 1, and this was statistically significant (for IL-4 p< 0.003, and for IL-5 p< 0.002). In group 3, both two cytokin levels were statistically lower than group 2 (for IL-4 p< 0.001, and for IL-5 p< 0.026). In BALF samples both two cytokin levels were higher in group 2 than group 1, and this was statistically significant (for IL-4 p< 0.004, and for IL-5 p< 0.001). In group 3, both two cytokin levels were lower than group 2, but it was not statistically significant (p> 0.05). In conclusion, it is thought that antiinflammatory effect of glucocorticoids occur by inhibiting the formation of IL-4, IL-5 and eosinophils. PMID- 16258884 TI - [Comparison of three clinical prediction rules among patients with suspected pulmonary embolism]. AB - Certain clinical findings raise the suspicion of pulmonary embolism (PE) and may be useful in selecting patients for further diagnostic testing. Three prediction rules for PE have been described recently: Wells' rule (WR), Geneva rule (GR) and Miniati' rule (MR). The aim of present study is to compare the predictive accuracy of the three methods on the basis of our patients' results. Eighty-five patients admitted to our department with suspicion of PE were included into the study. Sixty-three patients were discharged with the diagnosis of PE, whereas in 22 patients, the initial PE diagnosis was ruled out. The three methods for assessing the clinical probability of PE classified similar proportions of patients into the low, intermediate and high clinical probability categories. The frequencies of PE in each method (WR, GR and MR) were 5%, 64% and 14% in the low category, 90%, 80% and 75% in the intermediate category and 100%, 100% and 94% in the high category (p = 0.001, 0.064, 0.001) respectively. When we compared the performances of WR and GR, including all possible total score values, the area under the ROC curve (AUC) was 0.99 for the WR (p= 0.001) and 0.74 for the GR (p= 0.001). When we used only the three probability categories (low, intermediate, high), AUC was 0.96 for the WR (p= 0.001), 0.64 for the GR (p= 0.04), and 0.7 for the MR (p= 0.005). In conclusion, the present study indicates that clinical assessment is a fundamental step in the diagnostic work-up of PE. The Wells' method performs better than other two methods. PMID- 16258885 TI - [The diagnosis of definitive or probable tuberculosis and latent tuberculosis infection in children with suspected tuberculosis]. AB - Pulmonary tuberculosis (Tbc) continues to be an important cause of morbidity in children in our country and in the world. There are diagnostic difficulties in the evaluation of the patients with suspected Tbc. In our study of 118 cases, the signs or symptoms suggestive of pulmonary Tbc have a microbiologically confirmed ratio of 26% within the cases having a positive tuberculin test result or having a contact with an adult that had Tbc. Thirty-one (26%) patients were diagnosed with definite Tbc, 48 (41%) patients with probable Tbc and 28 (24%) patients with latent Tbc infection. The tuberculin test was positive in 22 (71%) patients with definite Tbc and in 29 (60%) patients with probable Tbc. A history of a contact with an adult having Tbc has been observed in 18 (58%) patients in definite Tbc case group, in 23 (48%) patients in probable Tbc case group and in 13 (46%) patients in latent Tbc infection case group. No significant difference has been observed between patients with definite or probable pulmonary Tbc according to the average age, sex, tuberculin test positivity and history of Tbc contact. The most common clinical symptoms that has been observed in both groups were cough, fever and weight loss and the most common radiological finding that has been found in both groups was persistent infiltration. Definite or probable Tbc cases were treated for a period of 6 months with 3 anti-Tbc drugs. There were no patients who had failed to response to the treatment. These results showed that, the diagnostic criteria used for probable pulmonary Tbc are also useful in detecting the patients who should be treated with anti-Tbc drugs. PMID- 16258886 TI - [Evaluation of phenol ammonium sulfate sedimentation method for diagnosis of pulmonary tuberculosis]. AB - In the study, the results of culture have been accepted as a gold standard and the specificity and sensitivity of phenol ammonium sulfate sedimentation method has been evaluated. When it is evaluated according to the results of culture, it has been found that the specificity and sensitivity of phenol ammonium sulfate sedimentation method is 90%, the specificity of the process which is made through the N-acetyl-L-cysteine NaOH method is 90% and the sensitivity of it is 85%. In conclusion, the phenol ammonium sulfate method seems to be as a secure method that can only be used in the laboratories in which microscopic studies are made. PMID- 16258887 TI - [Dust exposure levels and pneumoconiosis prevalence in a lignite coal miners]. AB - The effects of working conditions on health in lignite mines in Turkey have not been studied comprehensively. The aim of this study was to determine the prevalence of coal miner pneumoconiosis among lignite miners and to evaluate the effects of working conditions. The study was carried out between 2001 and 2003. All workers in the lignite mine and maintenance workers were consisted in the study. A face to face questionnaire was used to obtain demographics and work life variables. Posterior-anterior chest radiograms were evaluated by an A reader chest physician, according to ILO 1980 standard guidelines. Profusion of densities related with pneumoconiosis as 1/0 and above regarded as the "suspected pneumoconiosis". Parenchyma degeneration and/or massive opacities have been accepted as complicated silicosis. Dust concentrations in the work environment obtained from the periodical examinations. Two thousand four hundred and sixty four X-ray were evaluated and 333 (13.5%) pneumoconiosis compatible changes were found. Among the pneumoconiosis cases, 25 (7.5%) were assigned as complicated silicosis. There was significant and positive association between worked years and pneumoconiosis prevalence (p= 0.019). Our findings indicated that pneumoconiosis prevalence among lignite miners in Turkey comparable to the USA prevalence prior to implementation of effective dust control programmes. It has been suggested that dust exposure in the work environment were high enough to developed pneumoconiosis in lignite mines. Dust control systems and measures should be re-evaluated. PMID- 16258888 TI - [Poland syndrome (a case report)]. AB - Poland syndrome is characterized with unilateral absence of pectoralis major muscle. Its incidence is one in 30000 live births. A 20 years old case with Poland syndrome is presented together with its clinical and laboratory features in this study. The case had anomaly of shortness of right hand fingers and syndactily between second and third fingers in addition to absence of right pectoralis muscle group. There was not another associated anomaly except aforementioned ones. Strength loss in abduction and adduction of right shoulder was detected with Cybex dynamometer. Furthermore decrease in predicted maximal inspiratory and expiratory pressures was detected. PMID- 16258889 TI - A case of bronchogenic carcinoma presenting with acute abdomen. AB - Colonic metastasis of the bronchogenic carcinoma is quite rare. Here we document an extremely rare presentation of the lung cancer that presented with acute abdomen and was diagnosed as intestinal obstruction due to colon carcinoma initially. He underwent an urgent operation and the obliterating mass in the colon was resected and reported as "colon metastasis from epidermoid carcinoma probably of the lung". Afterwards bronchoscopy revealed an endobronchial lesion in the right lower lobe that was diagnosed as poorly differentiated squamous cell lung carcinoma. In this case, colon metastasis was diagnosed before the diagnosis of the primary disease. PMID- 16258890 TI - Gross mural cartilage in a congenital bronchogenic cyst: MRI features. AB - An infant who had a cardiac murmur after the delivery was evaluated with echocardiogram. A 1 cm cystic area was incidentally identified in the upper right mediastinum, that was not related with the heart. A 4 x 4.5 x 8 cm cystic mass in the superior mediastinum and lower neck was demonstrated on the magnetic resonance imaging (MRI). At surgery, a small solid structure corresponding to the nodule seen on MRI was identified, which was firm and cartilaginous in nature and attached to the wall of the cyst. Grossly visible cartilage in association with a bronchogenic cyst has not been previously reported. In conclusion, the bronchogenic cysts can have grossly visible cartilage in their wall and extend to extrathoracic spaces in a contiguous fashion, mimicking other entities such as teratomas and cystic lymphangiomas. PMID- 16258891 TI - [Churg-Strauss syndrome (two case reports)]. AB - Churg-Strauss syndrome is a necrotizing vasculitis with multiple organ involvement characterized by asthma, peripheral blood eosinophilia, eosinophilic tissue infiltration and extravascular granulomas. A 35 years-old male with 6 months history of asthma and a 43 years-old female with 4-years history of asthma, were further examined due to clinical worsening and lesions on chest radiographs. They were finally diagnosed as Churg-Strauss syndrome. Clinical and radiological response to oral corticosteroid therapy was obtained. PMID- 16258892 TI - [Pulmonary alveolar microlithiasis]. AB - Pulmonary alveolar microlithiasis (PAM) is a rare, chronic lung disease characterized by extensive intra alveolar calcium and phosphate deposition throughout both lung parenchyma. Etiology and pathogenesis of PAM is not known. There are some hypothesis for etiology and pathogenesis of PAM but none of them are satisfactory. The incidence of PAM is high in countries such as Turkey, Italy and USA. There is a surprising discordancy between radiological appearance and clinical presentation. Chest X-ray appearance of PAM is almost pathognomonic. Patients with PAM may have all findings of interstitial lung disease in varying degrees as well as micro-nodules on their HRCTs. Patients with PAM usually asymptomatic until the underlying process affects alveolar gas exchange however patients may ultimately develop hypoxemia and cor pulmonale. A definitive therapy for PAM did not exist. Patients with advanced lung disease may benefit from lung transplantation. PMID- 16258893 TI - [Obstructive sleep apnea syndrome, endothelial dysfunction and coronary atherosclerosis]. AB - In obstructive sleep apnea syndrome (OSAS), repetitive episodes of apnea cause increased sympathetic nerve activity, increased surges in arterial blood pressure, swings in intrathoracic pressure, oxidative stres, hypoxia and hypercapnia. The association of OSAS with some diseases, having endothelial dysfunction in their physiopathology, such as hypertension, diabetes mellitus, obesity, coronary artery diseases, stroke and heart failure is common. Increased sympathetic nerve activity and also endothelial dysfunction which are the results of hypoxia, have important roles in vascular complications of OSAS. When compared with healthy population, an important endothelial dysfunction in OSAS patients and relationship between OSAS severity and endothelial dysfunction have been shown. In this review, the relationship between OSAS and endothelial dysfunction was overviewed. PMID- 16258894 TI - [Diagnostic approach to solitary pulmonary nodule]. AB - The solitary pulmonary nodule is a common radiologic abnormality that is often detected incidentally. Many malignant and benign diseases can present as a solitary pulmonary nodule on a chest roentgenogram. It is important to differentiate malignant nodules from benign nodules in the least invasive way and to make as specific and an accurate diagnosis as possible. In this paper, a diagnostic approach to the solitary pulmonary nodule was discussed. PMID- 16258895 TI - LC-ESI-MS/MS analysis for the quantification of morphine, codeine, morphine-3 beta-D-glucuronide, morphine-6-beta-D-glucuronide, and codeine-6-beta-D glucuronide in human urine. AB - A liquid chromatographic-electrospray ionization-tandem mass spectrometric method for the quantification of the opiates morphine, codeine, and their metabolites morphine-3-beta-D-glucuronide (M-3-G), morphine-6-beta-D-glucuronide (M-6-G) and codeine-6-beta-D-glucuronide (C-6-G) in human urine has been developed and validated. Identification and quantification were based on the following transitions: 286 to 201 and 229 for morphine, 300 to 215 and 243 for codeine, 462 to 286 [corrected] for M-3-G, 462 to 286 for M-6-G, and 476 to 300 for C-6-G. Calibration by linear regression analysis utilized deuterated internal standards and a weighting factor of 1/X. The method was accurate and precise across a linear dynamic range of 25.0 to 4000.0 ng/ml. Pretreatment of urine specimens using solid phase extraction was sufficient to limit matrix suppression to less than 40% for all five analytes. The method proved to be suitable for the quantification of morphine, codeine, and their metabolites in urine specimens collected from opioid-dependent participants enrolled in a methadone maintenance program. PMID- 16258896 TI - Gas-phase tautomers of protonated 1-methylcytosine. Preparation, energetics, and dissociation mechanisms. AB - Tautomers of 1-methylcytosine that are protonated at N-3 (1+) and C-5 (2+) have been specifically synthesized in the gas phase and characterized by tandem mass spectrometry and quantum chemical calculations. Ion 1+ is the most stable tautomer in aqueous and methanol solution and is likely to be formed by electrospray ionization of 1-methylcytosine and transferred in the gas phase. Gas phase protonation of 1-methylcytosine produces a mixture of 1+ and the O-2 protonated tautomer (3+), which are nearly isoenergetic. Dissociative ionization of 6-ethyl-5,6-dihydro-1-methylcytosine selectively forms isomer 2+. Upon collisional activation, ions 1+ and 3+ dissociate by loss of ammonia and [C,H,N,O], whose mechanisms have been established by deuterium labeling and ab initio calculations. The main dissociations of 2+ following collisional activation are losses of CH2=C=NH and HN=C=O. The mechanisms of these dissociations have been elucidated by deuterium labeling and theoretical calculations. PMID- 16258898 TI - Molecular targets in melanoma: strategies and challenges for diagnosis and therapy. AB - In coming years, we expect rapid advances in cutaneous melanoma diagnosis and therapy, because of the incorporation of new technologies into experimental and clinical research. Major discoveries in melanoma are often made by investigators outside the field, and the melanoma research community will need to develop a better means of incorporating these advances into their work, to capitalize on the promise they hold for patients. A far greater level of cooperation between labs and clinics will be to bring new technology-based discoveries from bench-to bedside and back. Metastatic melanoma should become a treatable disease in the next few years, because specific inhibitors are expected for most major targets. However, major challenges lie ahead in securing funding, building infrastructure and gaining expertise in new technologies. To meet these challenges, multidisciplinary collaborations will be required all the more. PMID- 16258897 TI - Specificity of receptor-ligand interactions and their effect on dimerisation as observed by electrospray mass spectrometry: bile acids form stable adducts to the RXRalpha. AB - Electrospray (ES) mass spectrometry data is presented showing that agonist binding to the nuclear receptor (NR), retinoid X receptor alpha (RXRalpha), is competitive. The competitive nature of agonist binding can be used to discriminate between the specific and non-specific binding of small lipophilic molecules to NRs. Further, data is presented which show that high-affinity ligand binding to the RXRalpha ligand-binding domain (LBD) stabilises the domain homodimer. The results indicate that homodimerisation, a functional property of the receptor associated with the binding of agonist ligands, could be used to discriminate between specific and non-specific binding events. Additionally, we report on the remarkable stability of the gas-phase complex between the RXRalpha LBD protein and endogenous bile acids. Protein-bile acid interactions in the gas phase were found to be surprisingly strong, withstanding 'in-source' fragmentation in the ES interface, and, in the case of taurocholic acid (TCA) and lithocholic acid-3-sulphate (LCA-3-sulphate), collision-induced dissociation within the collision cell of a tandem mass spectrometer. Bile acids were found to be inactive towards RXRalpha in transfection assays, and have not been reported to be ligands for the RXRalpha, although lithocholic acid (LCA) has been found to be a competitor in the photoaffinity labelling of RXRbeta with 9-cis-retinoic acid (9-cis-RA). The observation of strong RXRalpha-bile acid non-covalent complexes in ES mass spectrometry highlight the danger of extrapolating gas-phase binding data to the solution phase and further to a possible biological activity, particularly when surface-active compounds such as bile acids are involved. The introduction of a competitive ligand-binding experiment can alleviate this problem and allow the differentiation between specific and non-specific binding. PMID- 16258899 TI - Clinical and radiographic outcomes of four different treatment strategies in patients with early rheumatoid arthritis (the BeSt study): a randomized, controlled trial. AB - OBJECTIVE: Several treatment strategies have proven value in the amelioration of rheumatoid arthritis (RA), but the optimal strategy for preventing long-term joint damage and functional decline is unclear. We undertook this study to compare clinical and radiographic outcomes of 4 different treatment strategies, with intense monitoring in all patients. METHODS: In a multicenter, randomized clinical trial, 508 patients were allocated to 1 of 4 treatment strategies: sequential disease-modifying antirheumatic drug monotherapy (group 1), step-up combination therapy (group 2), initial combination therapy with tapered high-dose prednisone (group 3), and initial combination therapy with the tumor necrosis factor antagonist infliximab (group 4). Treatment adjustments were made every 3 months in an effort to obtain low disease activity (a Disease Activity Score in 44 joints of < or =2.4). RESULTS: Initial combination therapy including either prednisone (group 3) or infliximab (group 4) resulted in earlier functional improvement than did sequential monotherapy (group 1) and step-up combination therapy (group 2), with mean scores at 3 months on the Dutch version of the Health Assessment Questionnaire (D-HAQ) of 1.0 in groups 1 and 2 and 0.6 in groups 3 and 4 (P < 0.001). After 1 year, mean D-HAQ scores were 0.7 in groups 1 and 2 and 0.5 in groups 3 and 4 (P = 0.009). The median increases in total Sharp/Van der Heijde radiographic joint score were 2.0, 2.5, 1.0, and 0.5 in groups 1-4, respectively (P < 0.001). There were no significant differences in the number of adverse events and withdrawals between the groups. CONCLUSION: In patients with early RA, initial combination therapy including either prednisone or infliximab resulted in earlier functional improvement and less radiographic damage after 1 year than did sequential monotherapy or step-up combination therapy. PMID- 16258900 TI - Involvement of subchondral bone marrow in rheumatoid arthritis: lymphoid neogenesis and in situ relationship to subchondral bone marrow osteoclast recruitment. AB - OBJECTIVE: To evaluate the presence and immunohistochemical characteristics of subchondral bone marrow inflammatory infiltrate in rheumatoid arthritis (RA) and to determine the in situ relationship between marrow inflammation and osteoclast recruitment. METHODS: Bone samples and paired synovia from 8 RA patients undergoing joint surgery were analyzed by immunohistochemistry and in situ hybridization for specific lymphoid neogenetic features, such as T and B cell composition, follicular dendritic cell (FDC) networks, peripheral lymph node addressin (PNAd)-positive high endothelial venules, and lymphoid chemokine expression. Osteoclasts were identified as multinucleated tartrate-resistant acid phosphatase (TRAP)-positive and cathepsin K-positive cells adherent to the bone surface. RESULTS: An inflammatory infiltrate with perivascular aggregates of variable size was detected in 7 (87.5%) of 8 synovial samples and in paired bone samples. Lymphoid neogenetic features typical of rheumatoid synovium were also recognized in the bone marrow. PNAd+ blood vessels were found in 4 of 8 patients, CD21+ FDC networks in 2 patients, CXCL13+ cells in 5 patients, and CCL21+ cells in 6 patients. TRAP-positive and cathepsin K-positive osteoclasts were identified on both the synovial and marrow sides of the bone surface. Bone marrow samples showing a higher degree of inflammation were characterized by a significantly increased number of osteoclasts adherent to the subchondral bone. CONCLUSION: Our data demonstrate that lymphoid aggregates with lymphoid neogenetic features are detectable on the subchondral side of the joint in established RA. Moreover, the local inflammation/aggregation process appears to be related to osteoclast differentiation on the marrow side of subchondral bone, supporting a functional role of the bone compartment in local damage. PMID- 16258901 TI - Circadian pattern of spontaneous behavior in monarthritic rats: a novel global approach to evaluation of chronic pain and treatment effectiveness. AB - OBJECTIVE: Preclinical evaluation is an essential step in the assessment of new antiinflammatory or analgesic drugs. This study was undertaken to develop a new mode of evaluation of drug effectiveness based on behavior indicating well-being in a rat model of chronic inflammatory pain. We chose to examine the circadian pattern of spontaneous behavior. METHODS: The work was performed with a model of chronic monarthritis induced by Freund's complete adjuvant. Variations in behavioral patterns during the time course of arthritis were analyzed. In a second phase, the impact of acetaminophen and 2 nonsteroidal antiinflammatory drugs (aspirin and celecoxib), which are currently used in clinical practice to treat chronic inflammation, was studied after 7 days of treatment. RESULTS: The nocturnal pattern of activity of healthy rats comprised 3 main bursts. Chronic painful monarthritis altered this spontaneous pattern of nocturnal behavior (normal period of activity). Monarthritic rats showed a decrease in the total time spent in activity during the night, and lost their pattern of activity. These behavioral disturbances were reversed after long-term treatment with acetaminophen or celecoxib, with celecoxib appearing to be more effective. Aspirin was ineffective. CONCLUSION: These results enabled us to test this new procedure as a means of assessing well-being or ill- being during stages of chronic inflammatory pain in rats, and the effectiveness of repeated pharmacologic treatments. PMID- 16258902 TI - Obesity is an independent contributor to functional capacity and inflammation in systemic lupus erythematosus. AB - OBJECTIVE: Obesity induces a proinflammatory state and is a major cause of morbidity in the general population. However, little is known about the effects of obesity in patients with chronic inflammatory illnesses such as systemic lupus erythematosus (SLE). METHODS: One hundred consecutive patients with SLE were studied to determine the relationship between body mass index (BMI) and functional capacity, measures of fatigue, quality of life, and the inflammation markers C-reactive protein (CRP), the erythrocyte sedimentation rate, and interleukin-6 (IL-6). The association between BMI and patient characteristics was determined, and multiple logistic regression models were used to adjust for age, sex, disease activity, and disease-related damage. RESULTS: Thirty-three patients had a normal BMI (< 25 kg/m(2)), 28 were overweight (25-29.9 kg/m(2)), and 39 were obese (> or =30 kg/m(2)). Obese patients had worse functional capacity, more fatigue, and higher concentrations of inflammation markers. The mean +/- SD modified Health Assessment Questionnaire (M-HAQ) score was 0.6 +/- 0.4 in obese patients compared with 0.3 +/- 0.4 and 0.2 +/- 0.3 in overweight patients and those with a normal BMI, respectively (P = 0.001). The mean +/- SD concentrations of CRP in obese patients (10.0 +/- 8.6 mg/liter) were higher than those in patients who were overweight (4.7 +/- 5.4 mg/liter) or had a normal BMI (6.2 +/- 9.9 mg/liter) (P < 0.001). Similarly, concentrations of IL-6 were higher in obese patients (P = 0.003). After adjusting for age, sex, disease activity, and damage indices, the associations between BMI and CRP (P < 0.001), M-HAQ scores (P = 0.005), and IL-6 concentrations (P = 0.01) remained significant. CONCLUSION: Obesity is independently associated with impaired functional capacity and inflammation markers in patients with lupus. Thus, weight loss may improve functional capacity and decrease cardiovascular risk factors. PMID- 16258903 TI - Unitary hypothesis for multiple triggers of the pain and strain of migraine. AB - Migraine headache is triggered by and associated with a variety of hormonal, emotional, nutritional, and physiological changes. The perception of migraine headache is formed when nociceptive signals originating in the meninges are conveyed to the somatosensory cortex through the trigeminal ganglion, medullary dorsal horn, and thalamus. Is there a common descending pathway accounting for the activation of meningeal nociceptors by different migraine triggers? We propose that different migraine triggers activate a wide variety of brain areas that impinge on parasympathetic neurons innervating the meninges. According to this hypothesis, migraine triggers such as perfume, stress, or awakening activate multiple hypothalamic, limbic, and cortical areas, all of which contain neurons that project to the preganglionic parasympathetic neurons in the superior salivatory nucleus (SSN). The SSN, in turn, activates postganglionic parasympathetic neurons in the sphenopalatine ganglion, resulting in vasodilation and local release of inflammatory molecules that activate meningeal nociceptors. Are there ascending pathways through which the trigeminovascular system can induce the wide variety of migraine symptoms? We propose that trigeminovascular projections from the medullary dorsal horn to selective areas in the midbrain, hypothalamus, amygdala, and basal forebrain are functionally positioned to produce migraine symptoms such as irritability, loss of appetite, fatigue, depression, or the quest for solitude. Bidirectional trafficking by which the trigeminovascular system can activate the same brain areas that have triggered its own activity in the first place provides an attractive network of perpetual feedback that drives a migraine attack for many hours and even days. PMID- 16258904 TI - Momentary relationship between cortisol secretion and symptoms in patients with fibromyalgia. AB - OBJECTIVE: To compare the momentary association between salivary cortisol levels and pain, fatigue, and stress symptoms in patients with fibromyalgia (FM), and to compare diurnal cycles of cortisol secretion in patients with FM and healthy control subjects in a naturalistic environment. METHODS: Twenty-eight patients with FM and 27 healthy control subjects completed assessments on salivary cortisol levels and pain, fatigue, and stress symptoms, 5 times a day for 2 consecutive days, while engaging in usual daily activities. Only those participants who adhered to the protocol (assessed via activity monitor) were included in the final analyses. RESULTS: Twenty FM patients and 16 healthy control subjects adhered to the protocol. There were no significant differences in cortisol levels or diurnal cortisol variation between FM patients and healthy controls. Among women with FM, a strong relationship between cortisol level and current pain symptoms was observed at the waking time point (t = 3.35, P = 0.008) and 1 hour after waking (t = 2.97, P = 0.011), but not at the later 3 time points. This association was not due to differences in age, number of symptoms of depression, or self-reported history of physical or sexual abuse. Cortisol levels alone explained 38% and 14% of the variation in pain at the waking and 1 hour time points, respectively. No relationship was observed between cortisol level and fatigue or stress symptoms at any of the 5 time points. CONCLUSION: Among women with FM, pain symptoms early in the day are associated with variations in function of the hypothalamic-pituitary-adrenal axis. PMID- 16258905 TI - Characterization and consequences of pain variability in individuals with fibromyalgia. AB - OBJECTIVE: A growing body of evidence suggests that real-time electronic assessments of pain are preferable to traditional paper-and-pencil measures. We used electronic assessment data derived from a study of patients with fibromyalgia (FM) to examine variability of pain over time and to investigate the implications of pain fluctuation in the context of a clinical trial. METHODS: The study group comprised 125 patients with FM who were enrolled in a randomized, placebo-controlled trial of milnacipran. Pain intensity levels were captured in real time by participants using electronic diaries. Variability in pain was assessed as the standard deviation of pain entries over time (pain variability index [PVI]). RESULTS: Substantial between-subject differences in pain variability were observed (mean +/- SD PVI 1.61 +/- 0.656 [range 0.27-4.05]). The fluctuation in pain report was constant over time within individuals (r = 0.664, P < 0.001). Individuals with greater variability were more likely to be classified as responders in a drug trial (odds ratio 6.14, P = 0.006); however, this association was primarily attributable to a greater change in pain scores in individuals receiving placebo (r = 0.460, P = 0.02) rather than active drug (r = 0.09, P > 0.10). CONCLUSION: Among individuals with FM, there were large between subject differences in real-time pain reports. Pain variability was relatively constant over time within individuals. Perhaps the most important finding is that individuals with larger pain fluctuations were more likely to respond to placebo. It is not clear whether these findings are applicable only to patients with FM or whether they may also be seen in patients with other chronic pain conditions. PMID- 16258906 TI - Smoking and HLA-DR shared epitope alleles in rheumatoid arthritis: comment on the article by Padyukov et al. PMID- 16258907 TI - The interaction of smoking and the HLA-DRB1 shared epitope in rheumatoid factor positive rheumatoid arthritis: comment on the article by Padyukov et al. PMID- 16258908 TI - Angiotensin receptor blockers ameliorate collagen-induced arthritis in mice: comment on the article by Sagawa et al. PMID- 16258909 TI - Skewed X chromosome inactivation in scleroderma: comment on the article by Ozbalkan et al. PMID- 16258914 TI - Hydroxyapatite crystals and rotator cuff disorders: comment on the article by Gomoll et al. PMID- 16258912 TI - Does oral glucosamine prevent the loss of proteoglycans in a rabbit model of osteoarthritis? Comment on the article by Tiraloche et al. PMID- 16258916 TI - Clinical images: Mechanical bull myopathy. PMID- 16258919 TI - Positive effects of moderate exercise on glycosaminoglycan content in knee cartilage: a four-month, randomized, controlled trial in patients at risk of osteoarthritis. AB - OBJECTIVE: To evaluate the effects of moderate exercise on glycosaminoglycan (GAG) content in knee cartilage in subjects at high risk of knee osteoarthritis (OA). METHODS: Forty-five subjects (16 women, mean age 46 years, mean body mass index 26.6 kg/m(2)) who underwent partial medial meniscus resection 3-5 years previously were randomized to undergo a regimen of supervised exercise 3 times weekly for 4 months or to a nonintervention control group. Cartilage GAG content, an important aspect of the biomechanical properties of cartilage, was estimated by delayed gadolinium-enhanced magnetic resonance imaging of cartilage (dGEMRIC), with results expressed as the change in the T1 relaxation time in the presence of Gd-DTPA (T1[Gd]). RESULTS: Thirty of 45 patients were examined by dGEMRIC at baseline and followup. The exercise group (n = 16) showed an improvement in the T1(Gd) compared with the control group (n = 14) (15 msec versus -15 msec; P = 0.036). To study the dose response, change in the T1(Gd) was assessed for correlation with self-reported change in physical activity level, and a strong correlation was found in the exercise group (n = 16, r(S) = 0.70, 95% confidence interval [95% CI] 0.31-0.89) and in the pooled group of all subjects (n = 30, r(S) = 0.74, 95% CI 0.52-0.87). CONCLUSION: This in vivo cartilage monitoring study in patients at risk of knee OA who begin exercising indicates that adult human articular cartilage has a potential to adapt to loading change. Moderate exercise may be a good treatment not only to improve joint symptoms and function, but also to improve the knee cartilage GAG content in patients at high risk of developing OA. PMID- 16258920 TI - PINCH-1 expression during early avian embryogenesis: implications for neural crest and heart development. AB - The invasion of the cardiac neural crest (CNC) into the outflow tract (OFT) and subsequent OFT septation are critical events during vertebrate heart development. We previously had performed four modified differential display (DD) screens in the chick embryo to identify genes that may be involved in CNC and heart development. Full-length sequence of one of the DD clones has been obtained and identified as chick PINCH-1. This particularly interesting new cysteine-histidine rich protein contains five protein-binding LIM domains (five double zinc fingers), a nuclear localization signal, and a nuclear export signal, allowing it to participate in integrin and growth factor signaling and possibly act as a transcription factor. We show here for the first time that chick PINCH-1 is expressed in neural crest cells, both in the neural fold and cardiac OFT, and is also expressed in mesoderm derived-structures, including the myocardium, during avian embryogenesis. The normal expression pattern and overexpression in neural crest cell explants suggest that PINCH-1 may be a regulator of neural crest cell adhesion and migration. PMID- 16258921 TI - A novel mutant phenotype implicates dicephalic in cyst formation in the Drosophila ovary. AB - The establishment of polarity in Drosophila requires the correct specification of the oocyte in early stages of oogenesis, its positioning at the posterior of the egg chamber, and signalling events between the oocyte and the adjacent posterior follicle cells. As a consequence, the anterior-posterior and the dorsal-ventral axes are fixed. The posterior localisation of the oocyte depends on cadherin mediated adhesion between the oocyte and the follicle cells. Here we show that dicephalic mutants affect the posterior positioning of the oocyte without interfering with oocyte specification in the germarium. Unlike other mutants that also affect the posterior placement of the oocyte, dicephalic mutants affect neither gurken expression nor karyosome formation during meiosis. By analysing in detail the mutant phenotypes of dicephalic, we find that cyst formation in mutant germaria is defective and that it shares some similarities with cysts that lack DE-cadherin in the germline cells. We propose a model in which dicephalic is involved in the proper adhesion between the oocyte and the somatic follicle cells. PMID- 16258922 TI - Viable human buccal mucosa cells do not yield typical nucleoids: impacts on the single-cell gel electrophoresis/Comet assay. AB - Buccal mucosa (BM) cells have been used in human biomonitoring studies for detecting DNA adducts and chromosomal damage in an epithelial cell population. In the present study, we have investigated if human BM cells are suitable for use in the single-cell gel electrophoresis (SCGE)/Comet assay as an approach for estimating the exposure of epithelial cells to DNA-damaging agents. Our results indicate that only a few cells from BM cell samples yield comets that can be analyzed by current methods, and that the yield of cells with comets is independent of the percentage of viable BM cells in the sample. Data generated after enzymatic enrichment of viable cells and immunomagnetic separation of epithelial cells suggest that most of the BM cells that do form comets are probably leukocytes. Moreover, by reevaluating specific cells after running the Comet assay, we found that viable epithelial BM cells give rise to atypical comets that are not included in the analysis. Comparing DNA migration patterns between small groups of smokers and nonsmokers indicated that long-term smoking had no effect on the subpopulation of cells that yield typical comets. Our results indicate that the SCGE assay, as it is commonly performed, may not be useful for genotoxicity monitoring in human epithelial BM cells. PMID- 16258924 TI - Translating advances in understanding of cardiovascular development into clinical care. PMID- 16258923 TI - A QSAR for the mutagenic potencies of twelve 2-amino-trimethylimidazopyridine isomers: structural, quantum chemical, and hydropathic factors. AB - An isomeric series of heterocyclic amines related to one found in heated muscle meats was investigated for properties that predict their measured mutagenic potency. Eleven of the 12 possible 2-amino-trimethylimidazopyridine (TMIP) isomers were tested for mutagenic potency in the Ames/Salmonella test with bacterial strain TA98, and resulted in a 600-fold range in potency. Structural, quantum chemical, and hydropathic data were calculated on the parent molecules and the corresponding nitrenium ions of all of the tested isomers to establish models for predicting the potency of the unknown isomer. The principal determinants of higher mutagenic potency in these amines are: (1) a small dipole moment, (2) the combination of b-face ring fusion and N3-methyl group, (3) a lower calculated energy of the pi electron system, (4) a smaller energy gap between the amine HOMO and LUMO orbitals (Pearson "softness"), and (5) a more stable nitrenium ion. Based on predicted potency from the average of six regression models, the isomer not yet synthesized and tested is expected to have a mutagenic potency of 0.77 revertants/microg in tester strain TA98, which is near the low end of the potency range of the isomers. PMID- 16258925 TI - Use of genetic toxicology information for risk assessment. AB - Genetic toxicology data are used worldwide in regulatory decision-making. On the 25th anniversary of Environmental and Molecular Mutagenesis, we think it is important to provide a brief overview of the currently available genetic toxicity tests and to outline a framework for conducting weight-of-the-evidence (WOE) evaluations that optimize the utility of genetic toxicology information for risk assessment. There are two major types of regulatory decisions made by agencies such as the Environmental Protection Agency (EPA) and the Food and Drug Administration (FDA): (1) the approval and registration of pesticides, pharmaceuticals, medical devices, and medical-use products, and (2) the setting of standards for acceptable exposure levels in air, water, and food. Genetic toxicology data are utilized for both of these regulatory decisions. The current default assumption for regulatory decisions is that chemicals that are shown to be genotoxic in standard tests are, in fact, capable of causing mutations in humans (in somatic and/or germ cells) and that they contribute to adverse health outcomes via a "genotoxic/mutagenic" mode of action (MOA). The new EPA Guidelines for Carcinogen Risk Assessment [Guidelines for Carcinogen Risk Assessment, USEPA, 2005, EPA Publication No. EPA/630/P-03/001F] emphasize the use of MOA information in risk assessment and provide a framework to help identify a possible mutagenic and/or nonmutagenic MOA for potential adverse effects. An analysis of the available genetic toxicity data is now, more than ever, a key component to consider in the derivation of an MOA for characterizing observed adverse health outcomes such as cancer. We provide our perspective and a two-step strategy for evaluating genotoxicity data for optimal use in regulatory decision-making. The strategy includes integration of all available information and provides, first, for a WOE analysis as to whether a chemical is a mutagen, and second, whether an adverse health outcome is mediated via a mutagenic MOA. PMID- 16258926 TI - Establishment and maintenance of planar epithelial cell polarity by asymmetric cadherin bridges: a computer model. AB - Animal scales, hairs, feathers, and cilia are oriented due to cell polarization in the epithelial plane. Genes involved have been identified, but the signal and mechanism remain unknown. In Drosophila wing polarization, the action of a gradient of Frizzled activity is widely assumed; and cell-cell signalling by cadherins such as Flamingo surely plays a major role. We present a computer model where reading the Frizzled gradient occurs through biased, feedback-reinforced formation of Flamingo-based asymmetric intercellular complexes. Through these complexes neighboring cells are able to compare their Frizzled activity levels. Our computations are highly noise-resistant and reproduce both wild-type and all known mutant wing phenotypes; other phenotypes are predicted. The model puts stringent limits on a Frizzled activation signal, which should exhibit unusual properties: (1) the extracellular Frizzled signalling gradient should be counterdirectional--decreasing from proximal (P) to distal (D), whereas during polarization, the intracellular Frizzled gradient builds up from P to D; (2) the external gradient should be relatively weak and short-lived, lest it prevent inversion of intracellular Frizzled. These features, largely independent of model details, may provide useful clues for future experimental efforts. PMID- 16258927 TI - Lessons learned from nine clinical trials of disease-modifying osteoarthritis drugs. PMID- 16258928 TI - Progress toward the cure of rheumatoid arthritis? The BeSt study. PMID- 16258929 TI - Adult cancer survivors: how are they faring? AB - This study identified the psychosocial problems that 752 patients from 3 states who had been diagnosed with 1 of the 10 most commonly occurring cancers indicated concerned them the most. Approximately 1 year after being diagnosed with cancer, 68.1% of patients were concerned with their illness returning, and more than half were concerned with developing a disease recurrence (59.8%) or had fears regarding the future (57.7%). In addition to these psychological problems focused on fear, approximately two-thirds (67.1%) of patients were concerned about a physical health problem, fatigue, and loss of strength. Two other physical health problems that concerned more than two-fifths of patients were sleep difficulties (47.9%) and sexual dysfunction (41.2%). More problems were reported by younger survivors (ages 18-54 yrs), women, nonwhites, those who were not married, and those with a household income of less than 20,000 US dollars a year. Those patients currently in treatment for cancer reported on average significantly more problems (P < 0.001) and on average had a higher Cancer Problems in Living Scale (CPILS) total score (P < 0.001) compared with those not currently in treatment. In a comparison of respondents with one of the four most common cancers, the most concerns regarding problems in living and highest mean CPILS scores were reported by those diagnosed with lung cancer, followed by survivors of breast cancer, colorectal cancer, and prostate cancer. PMID- 16258930 TI - Closing the divide: diverse populations and cancer survivorship. AB - Currently, there is a "divide" between cancer survivorship initiatives and minority participation. The level of participation is nearly nonexistent in many cancer support and control initiatives. Cancer survivorship resources that facilitate access to treatment, psychosocial interventions, clinical trials, and research are key components to eliminating this divide. Differences in cancer results among minority populations are caused by several factors, including biologic reactions to environmental activities, socioeconomic status, perceived beliefs and notions of medical professionals, a lack of resources to participate in cancer support groups, and having personal contact with cancer survivors. Health professionals, advocates, and researchers hold the key to opening more opportunities for the improvement of cancer survivorship among minorities. The belief that "one size fits all" is unrealistic. These beliefs can influence participation in innovative clinical trials, decisions about treatment, emotional responses, and social support relationships. To help ensure participation in these programs, researchers and health workers must understand the role of social and psychosocial implications and results of the assessment, strategies, and sustainability that must be included in the development stage of any cancer support and survivorship initiatives. For this article, the authors examined mechanisms that can be used by cancer-control researchers and program staff to limit the divide between cancer survivorship initiatives and minority participation. They identified three strategies that must be used to address this divide effectively: the inclusion of minorities in clinical trials, intervention studies, treatment, and research programs; the development of culturally sensitive environments; and the ability to sustain minority participation. In summary, cancer survivorship includes many components that are developed individually and collectively to formulate sound strategies for including minorities in cancer-control initiatives. These programs should go beyond basic support groups and should include research studies, clinical trials, and alternative treatments for increasing cancer survival rates and quality of life among minorities. The divide can be addressed only through a proactive initiative that brings cancer survivorship initiatives and minority communities together in full partnership. PMID- 16258931 TI - Pathways for psychosocial care of cancer survivors. AB - Today, the growing number of survivors from many sites of cancer necessitates that thought be given to ways that ensure follow-up psychosocial care and its integration into ongoing medical surveillance. The establishment of standards of care together with evidence- and consensus-based clinical practice guidelines have provided a highly effective method of enhancing quality care for treatment of cancer. There remain, however, major problems in dissemination and application of these guidelines on the clinical level. The National Comprehensive Cancer Network (NCCN) formed a Panel on Distress Management that developed the first set of consensus-based standards for psychosocial care and clinical practice guidelines specific to cancer illnesses. This article proposes the extension of their concepts to cancer survivors. A model is presented that can assist oncologists and multidisciplinary teams in busy ambulatory settings to more readily identify those survivors who are distressed, whose quality of life is impaired, and who may benefit from further psychological evaluation and treatment. Three groups of cancer survivors are identified for whom pathways for psychosocial care should be defined and developed: 1) survivors with physical sequelae, often resulting in significant neuropsychologic and physical consequences; 2) survivors with psychological sequelae or psychiatric disorders that interfere with functioning and quality of life; and 3) survivors with subsyndromal symptoms who have no identified physical or psychiatric sequelae, but who may nonetheless need help integrating the cancer experience into their lives to increase a sense of purpose, direction, and well being. A rapid screening tool for distress could be used at the time of follow-up visits to oncologists or physicians to identify patients with psychological, social, or spiritual concerns and could serve as a pathway for evaluation and referral for psychosocial counseling. Treating distress in these areas is to be viewed as an integral part of surveillance for survivors, and pathways to ensure integration are important. PMID- 16258932 TI - A model for care across the cancer continuum. AB - With contemporary therapy, the majority of children and adolescents who are diagnosed with cancer will be cured. However, curative therapy predisposes to adverse health outcomes that affect the long-term survivor's quality of life and increase the risk of early mortality. Recognition of the adverse effects of cancer treatment on growth and development, vital organ function, fertility and reproduction, and secondary carcinogenesis has been the stimulus for the development of risk-adapted treatment approaches for pediatric malignancies. Because the consequences of these therapeutic modifications may not manifest for many years, long-term follow-up is required to accurately define new or changing patterns of adverse health outcomes, appropriate screening approaches, and ultimately, risk-reducing interventions. Identification of vulnerable survivors is essential to provide timely interventions to detect, ameliorate, reduce, or prevent cancer-related sequelae. This process is challenging, because risk factors constantly are evolving as cancer therapies are modified and as survivors age. Aging also disrupts the continuity of after-cancer care, as adolescent and young adult survivors graduate from pediatric oncology practices to community medical providers who are largely unfamiliar with cancer-related health risks. Health outcomes research objectives that target cancer survivors must adapt as cancer therapies evolve and as new risk factors for cancer-related morbidity emerge. Prospectively using a multidimensional, comprehensive approach that considers host-related, cancer-related, genetic, and lifestyle factors in combination with results from focused medical and behavioral evaluations obtained from cancer survivors who participate in long-term follow-up programs provides an optimal method of defining high-risk profiles for adverse health outcomes across the age spectrum. PMID- 16258933 TI - Satellite cells from dystrophic (mdx) mice display accelerated differentiation in primary cultures and in isolated myofibers. AB - In the dystrophic (mdx) mouse, skeletal muscle undergoes cycles of degeneration and regeneration, and myogenic progenitors (satellite cells) show ongoing proliferation and differentiation at a time when counterpart cells in normal healthy muscle enter quiescence. However, it remains unclear whether this enhanced satellite cell activity is triggered solely by the muscle environment or is also governed by factors inherent in satellite cells. To obtain a better picture of myogenesis in dystrophic muscle, a direct cell-by-cell analysis was performed to compare satellite cell dynamics from mdx and normal (C57Bl/10) mice in two cell culture models. In one model, the kinetics of satellite cell differentiation was quantified in primary cell cultures from diaphragm and limb muscles by immunodetection of MyoD, myogenin, and MEF2. In mdx cell cultures, myogenin protein was expressed earlier than normal and was followed more rapidly by dual myogenin/MEF2A expression and myotube formation. In the second model, the dynamics of satellite cell myogenesis were investigated in cultured myofibers isolated from flexor digitorum brevis (FDB) muscle, which retain satellite cells in the native position. Consistent with primary cultures, satellite cells in mdx myofibers displayed earlier myogenin expression, as well as an enhanced number of myogenin-expressing satellite cells per myofiber compared to normal. The addition of fibroblast growth factor 2 (FGF2) led to an increase in the number of satellite cells expressing myogenin in normal and mdx myofibers. However, the extent of the FGF effect was more robust in mdx myofibers. Notably, many myonuclei in mdx myofibers were centralized, evidence of segmental regeneration; all central nuclei and many peripheral nuclei in mdx myofibers were positive for MEF2A. Results indicated that myogenic cells in dystrophic muscle display accelerated differentiation. Furthermore, the study demonstrated that FDB myofibers are an excellent model of the in vivo state of muscle, as they accurately represented the dystrophic phenotype. PMID- 16258934 TI - cadherin-6 message expression in the nervous system of developing zebrafish. AB - Cadherins are cell surface adhesion molecules that play important roles in development of a variety of tissues including the nervous system. In this study, we analyzed expression pattern of cadherin-6, a member of the type II cadherin subfamily, in the embryonic zebrafish nervous system using in situ hybridization methods. cadherin-6 message is first expressed by the neural keel, then by restricted regions in the brain and spinal cord. cadherin-6 expression in the brain transiently delineates specific brain regions. In the peripheral nervous system, cadherin-6 message is expressed by the neurogenic placodes and the dorsal root ganglia. As development proceeds, cadherin-6 expression domain and/or expression levels increased in the embryonic nervous system. Our results show that cadherin-6 expression in the zebrafish developing nervous system is both spatially and temporally regulated, implicating a role for cadherin-6 in the formation of these nervous structures. PMID- 16258935 TI - I-SceI meganuclease-mediated transgenesis in Xenopus. AB - Several experimental approaches have been described to generate transgenic frogs. Here, we report on the application of a novel method in Xenopus, making use of I SceI meganuclease. The characteristic feature of this endonuclease is that it has an extended recognition site of 18 bp, which is expected to exist only once in 7 x 10(10) bp of random DNA sequences. Various reporter constructs flanked by two I SceI recognition sites were injected together with the I-SceI meganuclease into one-cell stage Xenopus embryos. We observed an overall transgenesis frequency of 10% or more under optimized condition. The injected genes were integrated into the genome and transmitted to F1 offspring. Southern blot analysis showed that between one and eight copies of the transgene were integrated. Meganuclease-aided transgenesis, thus, provides a simple and highly efficient tool for transgenesis in Xenopus. PMID- 16258936 TI - Expression of avian C-terminal binding proteins (Ctbp1 and Ctbp2) during embryonic development. AB - C-terminal binding proteins (CtBPs) are transcriptional corepressors of mediators of Notch, Wnt, and other signalling pathways. Thus, they are potential players in the control of several developmentally important processes, including segmentation, somitogenesis, and neural tube and limb patterning. We have cloned the avian orthologues of Ctbp1 and Ctbp2 and examined their expression pattern by whole-mount in situ hybridization between Hamburger and Hamilton (HH) stages 3 and 24. Both Ctbp genes show similar expression patterns during embryonic development, and both are detected from HH stage 3 in the developing central nervous system, by HH stage 7 in the paraxial mesoderm and later in the limb bud. In most places, Ctbp1 and Ctbp2 are expressed in overlapping domains. However, there are interesting domains and/or temporal expression patterns that are specific to each Ctbp gene. For instance, Ctbp1 is predominantly expressed in the epiblast, whereas Ctbp2 is in the primitive streak at HH stage 3. However, by HH stage 4, both genes are found in the primitive streak and in the ectoderm. Similarly, although both genes display similar expression patterns in early somitogenesis, in mature somites, Ctbp1 transcripts are located in myotomal cells, whereas Ctbp2 transcripts are observed in dermomyotomal cells. Finally, we found that emigrating neural crest cells express Ctbp2, whereas dorsal root ganglia express Ctbp1. These data suggest that Ctbp1 and Ctbp2 may be functionally redundant in some tissues and have unique functions in other tissues. PMID- 16258937 TI - The 5' zebrafish scl promoter targets transcription to the brain, spinal cord, and hematopoietic and endothelial progenitors. AB - The stem cell leukemia (SCL) gene encodes a basic helix-loop-helix transcription factor and is essential for embryonic angiogenesis, hematopoietic stem cell specification, and erythrocyte maturation. Here, we report the isolation and characterization of the zebrafish scl promoter. We show that a 5-kilobase (kb) genomic fragment immediately upstream of the translation start site is capable of targeting the enhanced green fluorescence protein (EGFP) expression to the anterior and posterior lateral mesoderm where the endogenous scl normally expresses. Detailed analysis of the stable transgenic fish reveals that this 5-kb upstream sequence is sufficient to direct the EGFP transcription to the brain, spinal cord, and hematopoietic-endothelial progenitors, possibly the hemangioblast, but not primitive erythrocyte, suggesting that the zebrafish scl transcription in hematopoietic-endothelial progenitors and erythrocyte is regulated by distinct cis element(s). Our study has defined the cis regulatory element(s) for zebrafish scl expression in the brain, spinal cord, and hematopoietic-endothelial progenitors and established a valuable transgenic line Tg(5'5kbscl:EGFP) for studying hematopoietic lineage development. PMID- 16258938 TI - Expression patterns of Wnt genes during development of an anterior part of the chicken eye. AB - To address the roles of Wnts in the development of the anterior eye, we used a chicken model to perform comprehensive expression analysis of all Wnt genes during anterior eye development. In analyzing the available genomic sequences, we found that the chicken genome encodes 18 Wnt proteins that are homologous to corresponding human and mouse proteins. The mRNA sequences for 12 chicken Wnt genes are available in GenBank, and mRNAs for six other Wnt genes (Wnt2, Wnt5b, Wnt7b, Wnt8b, Wnt9b, and Wnt16) were identified and cloned based on the homology to the genes from other species. In addition, we found that chicken Wnt3a and Wnt7b genes encode two alternative mRNA isoforms containing different first exons. Following in situ hybridization, we found that out of 18 Wnt genes, 11 genes were expressed in the anterior eye, exhibiting distinct temporal-spatial patterns. Several Wnts were expressed in the lens, including Wnt2 and Wnt2b in the anterior epithelium and Wnt5a, Wnt5b, Wnt7a, and Wnt7b in the differentiating lens fiber cells. In the cornea, we detected Wnt3a, Wnt6, and Wnt9b in the ocular surface ectoderm, including the corneal epithelium, and Wnt9a in the corneal endothelium from the onset of its differentiation. In the optic cup, Wnt2, Wnt2b, and Wnt9a were localized in the rim of the optic cup (presumptive iris), while Wnt5a and Wnt16 were detected in the ciliary epithelium/iris zone of the differentiated optic cup, and Wnt6 was expressed in the iridial mesenchyme. These data suggest that Wnt signaling might play important roles in anterior eye development. PMID- 16258940 TI - Introduction: resilience of cancer survivors across the lifespan. PMID- 16258939 TI - Regulation of early Xenopus development by ErbB signaling. AB - ErbB signaling has long been implicated in cancer formation and progression and is shown to regulate cell division, migration, and death during tumorigenesis. The functions of the ErbB pathway during early vertebrate embryogenesis, however, are not well understood. Here we report characterization of ErbB activities during early frog development. Gain-of-function analyses show that EGFR, ErbB2, and ErbB4 induce ectopic tumor-like cell mass that contains increased numbers of mitotic cells. Both the muscle and the neural markers are expressed in these ectopic protrusions. ErbBs also induce mesodermal markers in ectodermal explants. Loss-of-function studies using carboxyl terminal-truncated dominant-negative ErbB receptors demonstrate that blocking ErbB signals leads to defective gastrulation movements and malformation of the embryonic axis with a reduction in the head structures in early frog embryos. These data, together with the observation that ErbBs are expressed early during frog embryogenesis, suggest that ErbBs regulate cell proliferation, movements, and embryonic patterning during early Xenopus development. PMID- 16258941 TI - Beat-to-beat blood pressure analysis after premature ventricular contraction indicates sensitive baroreceptor dysfunction in Parkinson's disease. AB - Extrasystoles occur in normal subjects but are significant more frequently (16.25% vs. 55%; chi(2) = 19.3; P < 0.001) seen in Parkinson's disease (PD) patients. The extrasystolic decreases in stroke volume and systolic pressure activate sympathetic vasomotor innervation and lead to a blood pressure increase for a few heartbeats. The purpose of this study was to prove whether the short time analysis of this blood pressure regulation allows the assessment of sympathetic neurocirculatory function. Records of noninvasive blood pressure monitoring were reviewed from 40 PD patients and 80 controls. A battery of cardiovascular autonomic tests, including Valsalva maneuver, tilt-table testing, echocardiography, and cardiac scintigraphy with [(123)I]meta-iodobenzylguanidine were performed. Fifty-five percent of the PD patients had at least one premature ventricular contraction (PVC) in 10 minutes lying supine at rest. After every PVC (13 PVCs) recorded from normal subjects, we found an increase in systolic blood pressure above base line with a maximum at the seventh heart beat. In all of the 22 PD patients, the systolic blood pressure was significantly decreased less than baseline in every PVC from the second to the ninth postextrasystolic beat (P < 0.001). In both groups, the extrasystolic fall in blood pressure was on average approximately 22%. The postextrasystolic potentiation did not differ (5.3% vs. 4.4%, not significant). If a PVC occurs, the analysis of short-time blood pressure regulation is a sensitive tool for baroreceptor reflex function. The advantage of this method results from the independence of patients cooperation and the high sensitivity to prove a sympathetic neurocirculatory failure within 10 heart beats. PMID- 16258942 TI - CYTE-I-HD: phase I dose finding and tolerability study of cysteamine (Cystagon) in Huntington's disease. AB - Cystamine, an inhibitor of transglutaminases, slows progression of Huntington's disease in the murine model by approximately 20%. Cysteamine, the dimer of cystamine, is an orphan drug approved for the treatment of nephropathic cystinosis and has a similar benefit in the murine model but with a narrower therapeutic window. In a single-center open-label study, we determined the maximum tolerable dose (MTD) and side effects of cysteamine in people with Huntington's disease. Cysteamine was started at a dose of 10 mg/kg per day, divided into four doses, and increased by 10 mg/kg per day weekly until the development of intolerable side effects or a maximum dose of 70 mg/kg per day. Of the 9 subjects, 1 had an MTD of 10 mg/kg per day, 1 had an MTD of 20 mg/kg per day, the maximum dose was 30 mg/kg per day for 2, 40 mg/kg per day for 2, and 50 mg/kg per day for 3. Dose-limiting side effects were motoric impairment in 5 and nausea in 4. The dose found tolerable by 8 of the subjects was 20 mg/kg per day. All had a noticeable hydrogen sulfide odor at doses of 40 mg/kg per day or higher. We conclude that, at a dose of 20 mg/kg per day, cysteamine was tolerable in people with Huntington's disease. Nausea and motoric impairment were the dose limiting side effects. PMID- 16258943 TI - Self-stimulatory behavior associated with deep brain stimulation in Parkinson's disease. PMID- 16258945 TI - Desmosomes in uterine epithelial cells decrease at the time of implantation: an ultrastructural and morphometric study. AB - Displacement of uterine epithelial cells is an important aspect of implantation in the rat and other species, allowing invasion of the blastocyst into the endometrial stroma. Desmosomes, which are part of the lateral junctional complex, function in cell-to-cell adhesion, and are therefore likely to be involved in displacement of uterine epithelial cells at the time of implantation. This study used transmission electron microscopy to study rat uterine epithelial cells during the peri-implantation period to investigate the change in the number of structural desmosomes along the lateral plasma membrane of uterine epithelial cells. We found a significant decrease in the number of desmosomes along the entire lateral plasma membrane as pregnancy progressed. Furthermore, there were also significant decreases in the number of desmosomes on the apical portion of the lateral plasma membrane between all days of pregnancy examined. In addition, on day 6 of pregnancy, the time of attachment, desmosomes were larger and seen as "giant desmosomes." For the first time, this study has shown that there is a significant reduction in cell height and actual number of ultrastructurally observable desmosomes at the time of implantation in the rat. It is proposed that this reduction in desmosome number leads to a decrease in lateral adhesion between uterine epithelial cells at the time of implantation, and hence is involved in the loss of uterine epithelial cells to facilitate blastocyst invasion. PMID- 16258944 TI - Cranial anatomy of the spade-headed amphisbaenian Diplometopon zarudnyi (Squamata, Amphisbaenia) based on high-resolution X-ray computed tomography. AB - The skull of the trogonophid amphisbaenian Diplometopon zarudnyi is described from high-resolution X-ray computed tomographic (HRXCT) imagery of a whole museum specimen preserved in ETOH. The skull was digitally resliced and disarticulated into individual elements, producing novel visualizations that allow detailed morphological analysis of its three-dimensionally complex structure. The prefrontal and jugal are absent in Diplometopon. The septomaxilla is present but hidden entirely from superficial view. In contrast to previous studies, we recognize a splenial fused to the compound bone of the mandible and a squamosal fused to the otic-occipital complex. Comparison of Diplometopon to the two other amphisbaenians previously described in comparable detail, Rhineura hatcherii and Amphisbaena alba, reveals a mosaic of cranial similarities and differences. Both Diplometopon and Rhineura exhibit a craniofacial angulation and expanded rostral blade related to use of the head as a digging tool, but the detailed architecture of these features is quite different. Additionally, whereas the snout of Rhineura exhibits a high degree of sculpturing and sensory innervation, this is not the case in Diplometopon. Unlike in Rhineura and Amphisbaena, the cranial elements of Diplometopon do not exhibit an extensive degree of overlap or complex interlocking sutures; instead, most of the cranial elements lie in loose apposition to each other. The degree to which this mosaic of features reflects functional demands, shared ancestry, and/or convergence is unclear in the absence of a stable hypothesis of amphisbaenian phylogeny. PMID- 16258946 TI - Cardiac conduction alterations in a French family with amyloidosis of the Finnish type with the p.Asp187Tyr mutation in the GSN gene. AB - Familial amyloidosis of the Finnish type (FAF) is a rare autosomal-dominant disorder caused by the accumulation of a 71-amino acid amyloidogenic fragment of mutant gelsolin, an actin-modulating protein. The main symptoms include corneal lattice dystrophy, progressive cranial and peripheral neuropathy, and skin changes. To date, only two mutations in the GSN gene have been described: the p.Asp187Asn mutation in most patients and the p.Asp187Tyr mutation in a Danish and Czech family. We report on the third family with the p.Asp187Tyr mutation and the first French FAF family. Severe cardiac conduction alterations in three patients were mainly caused by cardiac sympathetic denervation. These findings demonstrate the cardiological involvement of the FAF phenotype and suggest that cardiological follow-up is required in FAF patients. PMID- 16258947 TI - Abnormal expression of proteoglycans in Ullrich's disease with collagen VI deficiency. AB - Patients with Ullrich's disease have generalized muscle weakness, multiple contractures of the proximal joints, and hyperextensibility of the distal joints. Recently we found a marked reduction of fibronectin receptors in the skin and cultured fibroblasts of two patients with Ullrich's disease with collagen VI deficiency, and speculated that an abnormality of cell adhesion may be involved in the pathogenesis of the disease. In this study, we investigated the expression of proteoglycans and adhesion molecules in Ullrich's disease and other muscle diseases. We found a reduction of NG2 proteoglycan in the membrane of skeletal muscle but not in the skin in Ullrich's disease. By contrast, we found the upregulation of tenascin C in the extracellular matrix of skeletal muscle in Ullrich's disease. Our findings suggest that abnormal expression of proteoglycans and adhesion molecules may be involved in the pathogenesis of the dystrophic muscle changes in Ullrich's disease. PMID- 16258948 TI - Pulmonary predictors of survival in amyotrophic lateral sclerosis: use in clinical trial design. AB - The rapidity of progression of amyotrophic lateral sclerosis (ALS) to death or respiratory failure impacts patients, clinicians, and clinical investigators. This study compared the abilities of various pulmonary function tests to predict tracheostomy-free survival. We evaluated 95 ALS patients by determining upright and supine forced vital capacity (FVC), maximal inspiratory (MIP) and expiratory (MEP) pressures, arterial partial pressure of carbon dioxide (PaCO2), and transdiaphragmatic sniff pressures (Pdi-sniff). Tracheostomy-free survival time was measured from the date of spirometry. Supine FVC, upright FVC, MIP, MEP, and Pdi-sniff were significantly associated with tracheostomy-free survival after controlling for nonpulmonary factors, whereas PaCO2 was not. A normal supine FVC, MIP, or MEP was highly predictive for one-year survival. These tests are well suited to predict survival for trial enrollment and patient counseling. Supine FVC's simplicity of use and availability to ALS investigators makes it a particularly attractive predictor of one-year survival in ALS. PMID- 16258949 TI - Relapsing sensorimotor neuropathy with ophthalmoplegia, antidisialosyl antibodies, and extramembranous glomerulonephritis. AB - A 72-year-old man presented with oculomotor dysfunction, subacute relapsing sensorimotor neuropathy, elevated erythrocyte sedimentation rate, IgM monoclonal paraprotein, cold agglutinins, and antidisialosyl IgM antibodies, features previously described by the acronym CANOMAD (chronic ataxic neuropathy with ophthalmoplegia, M protein, agglutination, and disialosyl antibodies). The patient also had extramembranous glomerulopathy associated with this syndrome. Treatment with corticosteroids improved both the neuropathy and glomerulopathy. This case suggests that the spectrum of neuropathy associated with monoclonal gammopathy may be broader than originally believed. PMID- 16258950 TI - Differential response of heat shock proteins to hindlimb unloading and reloading in the soleus. AB - Hindlimb unloading (HU) results in oxidative stress, skeletal muscle atrophy, and increased damage upon reloading. Heat shock proteins (HSPs) protect against oxidative stress. However, it is unknown whether HSPs are depressed with long term unloading (28 days) or reloading. We tested the hypotheses that long-term HU would depress Hsp70 and Hsp25 pathways, whereas reloading would allow recovery in the soleus. Adult Sprague-Dawley rats were divided into three groups: controls; HU for 28 days; and HU + 7 days of reloading (HU-R). Soleus mass decreased with HU, and did not recover to control values with reloading. Hsp70 decreased with HU (-78.5%) and did not recover with HU-R (-81.4%). Upstream heat shock factor-1 was depressed with HU and HU-R. Hsp25 was reduced with HU, but recovered with reloading. Downstream of Hsp25, NADP-specific isocitrate dehydrogenase and glutathione peroxidase decreased with unloading, but only NADP-specific isocitrate dehydrogenase recovered with HU-R. Lipid peroxidation increased in both HU and HU-R. These data indicate that prolonged unloading and subsequent reloading results in complex, differential regulation of Hsp70 and Hsp25 pathways in the rat soleus muscle. Thus dysregulation and uncoupling of the Hsp70 and Hsp25 pathways may lead not only to muscle atrophy with prolonged unloading, but also impaired recovery of muscle mass during early reloading. PMID- 16258951 TI - Prenatal forehead edema in 4p- deletion: the 'Greek warrior helmet' profile revisited. AB - OBJECTIVES: Deletion of short arm of chromosome 4 is difficult to ascertain prenatally, and can be missed. METHODS: A prenatal suspicion of 4p- syndrome was thoroughly investigated by using two-dimensional and three-dimensional sonography, with a description of the fetal face dysmorphological pattern. The cytogenetic confirmation, obtained by karyotype and FISH technique, allowed a precise description of the prenatal abnormalities. Post-termination tridimensional helicoidal scanner of the fetal face was performed. RESULTS: The main anomaly discovered using two-dimensional sonography was the presence of a strikingly thick prefrontal edema (8 mm, twice the normal values, at 22 weeks: 3.81 +/- 0.62 mm). Three-dimensional sonography showed the classical postnatal profile, with the phenotypic aspect of a 'Greek warrior helmet'. Nasal bones were normal in size and placement, confirmed by helicoidal scanner. CONCLUSION: Prenatal diagnosis of 4p deletion syndrome can be difficult, and it is the presence of prefrontal edema, associated with more subtle facial anomalies (short philtrum, microretrognathia) which should trigger cytogenetic investigation for 4p- deletion, even with only borderline growth retardation. PMID- 16258952 TI - Variceal hemorrhage and cystic fibrosis: outcomes and implications for liver transplantation. AB - Autopsy and imaging studies show that liver involvement is common in cystic fibrosis. However, complications of chronic liver disease including portal hypertension and variceal bleeding are infrequently encountered, and the degree to which variceal hemorrhage affects prognosis in cystic fibrosis is unclear. This uncertainty has lead to debate as to whether liver transplantation is indicated in these patients. We describe a case series of 18 patients and compare their survival with a control group of cystic fibrosis patients without liver disease. The median age at first bleed was 20.0 years (range 9.7-30.9). The median survival after first bleed was 8.4 years, compared to 13.0 years in the control group (P = 0.15). A total of 14 patients have died, 9 from respiratory disease with no discernable contribution from their liver disease. Liver disease contributed to 4 deaths. Only 1 patient suffered a fatal hemorrhage, which may have been either variceal or bronchial in origin. Long-term survival is a frequent occurrence in patients with cystic fibrosis who suffer variceal hemorrhage, and age at death is comparable to the general cystic fibrosis population. In conclusion, this suggests that liver transplantation is not indicated in these patients without additional features of liver decompensation. PMID- 16258953 TI - High preoperative recipient plasma 7beta-hydroxycholesterol is associated with initial poor graft function after liver transplantation. AB - Oxidative stress is implicated in the pathogenesis of hepatic ischemia reperfusion injury, a major determinant of initial poor graft function (IPGF) after orthotopic liver transplantation (OLT). We prospectively investigated the association between the recipient plasma preoperative oxidative stress and the occurrence of IPGF after deceased-donor OLT and indirectly studied the source hepatic or extra-hepatic-of systemic oxidative stress in vivo in cirrhosis. We used a recently developed specific and sensitive mass spectrometry assay to measure 7beta-hydroxycholesterol and 7-ketocholesterol (oxysterols), markers of oxidative stress, in biological matrices. At univariate analysis, preoperative recipient 7beta-hydroxycholesterol plasma concentration was significantly higher in transplants with subsequent IPGF (n = 9) compared with those with initial good graft function (IGGF; n = 23) [mean +/- SD: 30.63 +/- 26.42 and 11.57 +/- 15.76 ng/mL, respectively] (P = 0.017). In a logistic regression model, which included also the Model for End-Stage Liver Disease (MELD) score, 7beta-hydroxycholesterol plasma concentration was an independent predictor of IPGF with an odds ratio of 1.17 (95% CI, 1.02-1.33, P = 0.028). Patients with cirrhosis (n = 32) had increased oxysterol plasma levels compared with healthy controls (n = 49); livers with cirrhosis (n = 21), however, had oxysterol content comparable with normal livers obtained from organ donors (n = 19). Oxysterols persisted elevated in plasma 1 month after OLT (n = 23). In conclusion, cirrhosis presents upregulated systemic oxidative stress likely of extrahepatic source that is associated with graft failure after OLT. PMID- 16258954 TI - Fine allelotyping of Erbb2-induced mammary tumors in mice reveals multiple discontinuous candidate regions of tumor-suppressor loci. AB - Loss of heterozygosity (LOH) at human chromosome bands 1p32-36 and 10q23-26 is frequent in various human tumors, including breast cancers, and is thought to reflect the loss of tumor-suppressor genes (TSGs). To map such genes, high resolution LOH analysis was performed on 93 Erbb2-induced mammary tumors from (BALB/c x C57BL/6) F1 MMTV/Erbb2 transgenic mice. A panel of 24 microsatellite markers specific to the region of mouse chr4, homologous to human 1p31-36, and 16 markers specific to the mouse chr19 region, homologous to human 10q23-26 were used. In addition, lower-density mapping was performed on the remaining portion of mouse chr4 [homologous to human 9p13, 9p21-24, 9q21-22, 9q31-34 (12 markers)] and chr19 [homologous to 9q21, 9p24, 11q12-13 (9 markers)]. Several distinct, discrete, and discontinuous LOH regions flanked by areas of heterozygosity were identified, 22 on chr4 and 14 on chr19. Among these, 13 were mapped in the region of homology with human 1p31-36 (between D4Mit153 and D4Mit254) and 9 in the region of homology with human 10q23-26 (between D19Mit46 and D19Mit6). Although several LOH loci span a large interval, many are relatively short (1-4 Mb), and a few span an interval of <1 Mb. This allelotyping represents the highest density of LOH loci yet mapped in these chromosomal regions. The presence of numerous LOH regions in alternation with regions of heterozygosity, consistent with mitotic recombination as a mechanism for generating such a mosaic pattern, suggests the presence of several TSGs in these regions and should facilitate their identification. PMID- 16258955 TI - Gross SDHB deletions in patients with paraganglioma detected by multiplex PCR: a possible hot spot? AB - Pheochromocytoma and paraganglioma are rare neuroendocrine tumors that arise in the adrenal medulla and the extra-adrenal paraganglia, respectively. Inheritance of these tumors is mainly a result of mutations affecting the VHL, RET, NF1, and SDH genes. Germ-line mutations of the SDH genes have been found to account for nearly 10% of apparently sporadic cases. Nevertheless, alterations other than point mutations have not yet been well characterized. In this study, we investigated the frequency of gross SDH deletions in 24 patients who tested negative for point mutations and had at least one of the recommended features for genetic testing. For this purpose, we used a technique that is easy to implement in the lab to specifically detect gross deletions affecting SDHB, SDHC, and SDHD. We identified 3 heterozygous SDHB deletions (3/24) in 3 independent cases with paraganglioma: 1 whole SDHB deletion and 2 deletions exclusively affecting exon 1. These latter mutations match the unique gross deletion previously reported, indicating this region could be a hot spot for gross SDHB deletions. It seems likely that these alterations can account for a considerable number of both familial and apparently sporadic paraganglioma cases. Although this is the first report describing the presence of gross deletions in patients with apparently sporadic paragangliomas, the extra-adrenal location of the tumor seems to constitute a determining factor for whether to include these patients in genetic testing for gross deletions in the SDHB gene. PMID- 16258956 TI - Differential effect of epigenetic alterations and genomic deletions of CDK inhibitors [p16(INK4a), p15(INK4b), p14(ARF)] in mantle cell lymphoma. AB - Mantle cell lymphoma (MCL) is characterized by the chromosomal translocation t(11;14)(q13;q32), resulting in overexpression of CCND1 in the vast majority of cases. In addition, alterations of other cell-cycle-regulating signal pathways (CDKN2B/CDKN2A-CCND1 and ARF-MDM2-TP53) are frequently observed. However, the hierarchy of promoter methylations and genomic alterations as well as the interaction with other cell-cycle regulator CDKN1A is poorly understood. A complete methylation-specific PCR coupled with direct sequencing of 71 MCL patient samples previously characterized for TP53 alterations, Ki67 expression by immunohistochemistry, and other genomic alterations was performed. In contrast to rare p16(INK4a) promoter methylation (9%), frequent p15(INK4b) (62%) and p14(ARF) (70%) promoter methylation was detectable in MCL. In an additional 16% of MCL cases, LOH for p16(INK4a) was detected. However, MCL cases with p15(INK4b) methylation tended to have lower proliferation (73% vs. 57%), and p15(INK4b) and p14(ARF) promoter methylation was also detected in normal stem cells. Therefore, epigenetic changes of those genes seem not to represent primary oncogenic mechanisms but physiological mechanisms of cell regulation. The rare p16(INK4a) promoter methylation and p16(INK4a) genetic alterations were directly correlated to cell proliferation and therefore are regarded as additional molecular alterations involved in the cell-cycle dysregulation of MCL. PMID- 16258957 TI - Influence of salivary organic substances on the discoloration of esthetic dental materials-a review. AB - The objective of this article was to review the articles on the interaction of salivary organic substances with resin-based dental materials and on the interaction of these organic substances with exogenous chemical agents, which results in discoloration. Original scientific articles or reviews on the saliva, acquired pellicle, and the interaction with pellicle and chemical agents related to dental resin-based materials were reviewed. Salivary esterases can increase or decrease the internal and external discoloration. The formation of acquired pellicle on the surface of a material varies by the properties of material, and the pellicle interacts with denaturation agents, such as tannin and chlorhexidine, to form stains and also adsorbs staining substances. Therefore, for the quality and longevity of restorations, protocols for the evaluation of the influence of organic substances on the extrinsic staining of restorative materials should be included in the evaluation of aesthetic restorative materials. PMID- 16258958 TI - Manufacturing, mechanical characterization, and in vitro performance of bioactive glass 13-93 fibers. AB - Fibers were manufactured from the bioactive glass 13-93 by melt spinning. The fibers were further characterized by measuring their tensile and flexural strength, and their in vitro performance was characterized by immersing them in simulated body fluid, which analyzed changes in their mass, their flexural strength, and surface reactions. The strength of glass fibers is highly dependent on fiber diameter, test method, and possible surface flaws, for example, cracks due to abrasion. In this study, the thinnest fibers (diameter between 24 and 33 microm) possessed the highest average tensile strength of 861 MPa. The flexural strength was initially 1353.5 MPa and it remained at that level for 2 weeks. The Weibull modulus for both tensile and flexural strength values was initially about 2.1. The flexural strength started to decrease and was only approximately 20% of the initial strength after 5 weeks. During the weeks 5-40, only a slight decrease was detected. The flexural modulus decreased steadily from 68 to 40 GPa during this period. The weight of the samples initially decreased due to leaching of ions and further started to increase due to precipitation of calcium phosphate on the fiber surfaces. The mass change of the bioactive glass fibers was dependent on the surface area rather than initial weight of the sample. The compositional analysis of the fiber surface after 24 h and 5 weeks immersion did confirm the initial leaching of ions and later the precipitation of a calcium phosphate layer on the bioactive glass 13-93 fiber surface in vitro. PMID- 16258959 TI - Poly-epsilon-caprolactone/hydroxyapatite composites for bone regeneration: in vitro characterization and human osteoblast response. AB - Polycaprolactone (PCL), a semicrystalline linear resorbable aliphatic polyester, is a good candidate as a scaffold for bone tissue engineering, due to its biocompatibility and biodegradability. However, the poor mechanical properties of PCL impair its use as scaffold for hard tissue regeneration, unless mechanical reinforcement is provided. To enhance mechanical properties and promote osteoconductivity, hydroxyapatite (HA) particles were added to the PCL matrix: three PCL-based composites with different volume ratio of HA (13%, 20%, and 32%) were studied. Mechanical properties and structure were analysed, along with biocompatibility and osteoconductivity. The addition of HA particles (in particular in the range of 20% and 32%) led to a significant improvement in mechanical performance (e.g., elastic modulus) of scaffold. Saos-2 cells and osteoblasts from human trabecular bone (hOB) retrieved during total hip replacement surgery were seeded onto 3D PCL samples for 1-4 weeks. Following the assessment of cell viability, proliferation, morphology, and ALP release, HA loaded PCL was found to improve osteoconduction compared to the PCL alone. The results indicated that PCL represents a potential candidate as an efficient substrate for bone substitution through an accurate balance between structural/ mechanical properties of polymer and biological activities. PMID- 16258960 TI - Buried alive: how osteoblasts become osteocytes. AB - During osteogenesis, osteoblasts lay down osteoid and transform into osteocytes embedded in mineralized bone matrix. Despite the fact that osteocytes are the most abundant cellular component of bone, little is known about the process of osteoblast-to-osteocyte transformation. What is known is that osteoblasts undergo a number of changes during this transformation, yet retain their connections to preosteoblasts and osteocytes. This review explores the osteoblast-to-osteocyte transformation during intramembranous ossification from both morphological and molecular perspectives. We investigate how these data support five schemes that describe how an osteoblast could become entrapped in the bone matrix (in mammals) and suggest one of the five scenarios that best fits as a model. Those osteoblasts on the bone surface that are destined for burial and destined to become osteocytes slow down matrix production compared to neighbouring osteoblasts, which continue to produce bone matrix. That is, cells that continue to produce matrix actively bury cells producing less or no new bone matrix (passive burial). We summarize which morphological and molecular changes could be used as characters (or markers) to follow the transformation process. PMID- 16258961 TI - Preparation and swelling behavior of chitosan-based superporous hydrogels for gastric retention application. AB - Chitosan and glycol chitosan hydrogels were prepared, and their swelling behaviors in acidic solution were studied to investigate their application for gastric retention device. The optimum preparation condition of superporous hydrogels was obtained from the gelation and blowing kinetics measured at varying acidic conditions. Both the swelling rate and swelling ratio of glycol chitosan hydrogels were higher than those of chitosan hydrogels. Swelling behaviors were significantly affected by not only foaming/drying methods but also crosslinking density, as the sizes and structures of pores generated were highly dependent on those preparation conditions. The prepared superporous hydrogels were highly sensitive to pH of swelling media, and showed reversible swelling and de-swelling behaviors maintaining their mechanical stability. The degradation kinetics in simulated gastric fluid was also studied. PMID- 16258962 TI - Synthesis and characterization of poly-alpha,beta-[N-(2-hydroxyethyl)-L aspartamide]-g-poly(L-lactide) biodegradable copolymers as drug carriers. AB - A series of biodegradable amphiphilic graft polymers were successfully synthesized by grafting poly(L-lactide) (PLLA) sequences onto a water-soluble polymer poly-alpha,beta-[N-(2-hydroxyethyl)-L-aspartamide] (PHEA) backbone. We established the feasibility of preparing these novel graft polymers by the ring opening polymerization initiated by the macroinitiator PHEA bearing hydroxyl groups without adding any catalyst. The successful grafting of PLLA sequences onto the PHEA backbone was verified by combined size exclusion chromatography (SEC) and multiangle laser light scattering (MALLS) analysis. The chemical structures of graft polymers were characterized by FTIR and (1)H NMR. The critical micelle concentration (CMC) of the graft polymer was determined by fluorescence probe technique using pyrene. By controlling the feed ratio of the macroinitiator to the monomer, graft polymers with different branch lengths can be obtained. Using the 3-(4,5 dimethylthiozol-2-yl)-2,5-diphenyl-tetrazolium bromide (MTT) assay, the graft copolymer has been proved to have low cytotoxicity. Based on the amphiphilicity of the graft copolymers, nanoparticular drug delivery systems were prepared by the direct dissolution method and the dialysis method. The anticancer drug Tegafur was encapsulated into polymeric nanoparticles, and in vitro drug release behavior was investigated. Transmission electron microscopy (TEM) images demonstrate that these nanoparticles are regularly spherical in shape. The particle size and distribution of the nanoparticles were measured. PMID- 16258963 TI - Epithelial transformations in the establishment of the blood-gas barrier in the developing chick embryo lung. AB - The tall epithelium of the developing chick embryo lung is converted to a squamous one, which participates in formation of the thin blood-gas barrier. We show that this conversion occurred through processes resembling exocrine secretion. Initially, cells formed intraluminal protrusions (aposomes), and then transcellular double membranes were established. Gaps between the membranes opened, thus, severing the aposome from the cell. Alternatively, aposomes were squeezed out by adjacent cells or were spontaneously constricted and extruded. As a third mechanism, formation and fusion of severed vesicles or vacuoles below the aposome and their fusion with the apicolateral plasma membrane resulted in severing of the aposome. The atria started to form by progressive epithelial attenuation and subsequent invasion of the surrounding mesenchyme at regions delineated by subepithelial alpha-smooth muscle actin-positive cells. Further epithelial attenuation was achieved by vacuolation; rupture of such vacuoles with resultant numerous microfolds and microvilli, which were abscised to accomplish a smooth squamous epithelium just before hatching. PMID- 16258964 TI - Spatio-temporal expression of MRF4 transcripts and protein during Xenopus laevis embryogenesis. AB - Whereas there have been extensive studies of the expression of XMyf5 and XMyoD during Xenopus embryogenesis, nothing is known about the spatio-temporal accumulation of XMRF4 transcripts and protein. In this report, we describe the cloning and characterization of two full-length MRF4 cDNAs and of their proximal promoters in Xenopus laevis. The comparison of the relative transcript levels of the XMRF4-a and -b genes in developing and adult muscles is highly suggestive of specific functions for the corresponding XMRF4 proteins. Whole-mount embryo in situ hybridization revealed the first XMRF4 transcripts in the more differentiated anterior myocytes of the embryo when the myosin heavy chain E3 mRNA begins to be detectable. XMRF4 mRNA accumulation later extended posteriorly but was never detected in the posterior unsegmented mesoderm, in contrast to XMyoD and XMyf-5. Whole-mount embryo immunohistochemistry revealed that XMRF4 protein accumulated in somite nuclei slightly after XMRF4 transcripts. PMID- 16258965 TI - Transforming growth factor-beta induces loss of epithelial character and smooth muscle cell differentiation in epicardial cells. AB - During embryogenesis, epicardial cells undergo epithelial-mesenchymal transformation (EMT), invade the myocardium, and differentiate into components of the coronary vasculature, including smooth muscle cells. We tested the hypothesis that transforming growth factor-beta (TGFbeta) stimulates EMT and smooth muscle differentiation of epicardial cells. In epicardial explants, TGFbeta1 and TGFbeta2 induce loss of epithelial morphology, cytokeratin, and membrane associated Zonula Occludens-1 and increase the smooth muscle markers calponin and caldesmon. Inhibition of activin receptor-like kinase (ALK) 5 blocks these effects, whereas constitutively active (ca) ALK5 increases cell invasion by 42%. Overexpression of Smad 3 did not mimic the effects of caALK5. Inhibition of p160 rho kinase or p38 MAP kinase prevented the loss of epithelial morphology in response to TGFbeta, whereas only inhibition of p160 rho kinase blocked TGFbeta stimulated caldesmon expression. These data demonstrate that TGFbeta stimulates loss of epithelial character and smooth muscle differentiation in epicardial cells by means of a mechanism that requires ALK5 and p160 rho kinase. PMID- 16258966 TI - Wingless signaling in a large insect, the blowfly Lucilia sericata: a beautiful example of evolutionary developmental biology. AB - Blowflies are the primary facultative agent in causing myiasis of domestic sheep in the whole world and, at the same time, it is an important tool for forensic medicine. Surprisingly, and in contrast to its importance, almost no data regarding the embryology and molecular markers are known for this insect. In this report, we present a detailed description of the blowfly Lucilia sericata embryogenesis and of imaginal disc development. The embryogenesis of Lucilia strongly resembles that of Drosophila, despite their apparent size difference. Moreover, imaginal disc development appears to be equally well conserved. Through cloning, expression, and functional studies, we show that the Lucilia Wingless (Wg) protein is highly conserved between the two species. We further show that parasegments are established in Lucilia, however, engrailed expression shows a more dynamic expression pattern than expected in comparison to Drosophila. Over expression of Lucilia Wingless in Drosophila shows wingless-like wing phenotypes, suggesting that Lucilia Wingless blocks the signalling activity of Drosophila Wingless. Upon injection of wg dsRNA, we observe a "lawn of denticle" phenotype, closely resembling that of Drosophila. Due to the large size of the insect, the distance over which Wingless exerts signalling activity is up to three times larger than in Drosophila, yet the consequences are very similar. Our data demonstrate long-range wingless signaling mechanisms adapted for patterning large domains of naked cuticle and suggest signaling properties of Lucilia Wingless that are distinct from those of Drosophila Wingless. PMID- 16258967 TI - Klippel-Trenaunay syndrome associated with polydactyly. AB - Klippel-Trenaunay syndrome (KTS) manifests cutaneous vascular nevus, superficial venous varicosities, and hypertrophy of the affected limb. KTS may be associated with other developmental anomalies such as polydactyly, syndactyly, and macrocephaly. We present a case with KTS associated with polydactyly. PMID- 16258968 TI - Bilateral thoracic ducts with coexistent persistent left superior vena cava. AB - A case of bilateral thoracic ducts with coexistent persistent left superior vena cava (SVC) was identified in a 77-year-old Japanese female cadaver during dissection in a gross anatomy course. The persistent left SVC began at the lower surface of the left brachiocephalic vein, descended in front of the aortic arch, and drained into the right atrium through the coronary sinus. The right SVC was normal both in size and in position. The azygos vein, receiving the hemiazygos vein, opened into the right SVC. The accessory hemiazygos vein and the left superior intercostal vein united to form a common trunk, which drained into the left SVC. The left and right thoracic ducts began at the level of the 1st lumbar vertebra, ran upwards parallel and anterior to the vertebral column, and terminated at the venous angles of their corresponding sides. There was an anastomotic branch between them. The present case was considered to be very rare, since the persistent left SVC and bilateral thoracic ducts coexisted. The embryologic basis and clinical importance of this case are discussed. PMID- 16258969 TI - Right coronary ostium agenesis with absence of the right coronary artery: a rare case of non-ST elevation coronary syndrome. AB - We present a case with right coronary ostium agenesis with anomalous origin of the right coronary artery from the left circumflex artery, which caused a non-ST elevation coronary syndrome. A review of the literature indicates this to be an extremely rare case. PMID- 16258970 TI - Charles Estienne (Carolus Stephanus) (ca.1504-1564): physician and anatomist. PMID- 16258971 TI - Correlation of magnetic resonance imaging with histopathology in arrhythmogenic right ventricular dysplasia/cardiomyopathy (ARVD/C). AB - Arrhythmogenic right ventricular dysplasia/cardiomyopathy (ARVD/C) is an asymptomatic cardiac disease characterized by fatty infiltration of the right ventricular myocardium and often results in sudden cardiac death. ARVD/C diagnosis includes the assessment of fatty infiltration, which can be achieved noninvasively with cardiovascular magnetic resonance (CMR). The standard CMR protocol places the signal-generating coil directly on the anterior chest wall and produces a nonspecific high intensity signal that obscures the high signal from fatty infiltration. The aim of this study was to determine whether increasing the coil-to-chest distance would improve identification of fatty infiltration. Thoraces from seven embalmed cadavers were imaged on a conventional 1.5 Tesla CMR scanner using the control protocol and an experimental protocol, with a 6 cm coil-to-chest distance. A representative axial MR image and corresponding gross section of the heart were analyzed in each case. Fatty infiltration was graded in a blinded fashion on the MR images with independent histopathologic assessment. In five of the seven cases, the experimental protocol provided a correlation between CMR and histopathology that was as good as or better than the control protocol. The experimental protocol was also better in preventing false positive diagnosis in cases of minimal infiltration. Thus, the experimental protocol showed a stronger correlation with histopathology than did the control protocol. Increasing the distance between the anterior surface coil and chest wall may improve classification of presence or absence of fatty infiltration in the right ventricular myocardium, potentially improving the noninvasive detection of ARVD/C with CMR. PMID- 16258972 TI - The course of the greater occipital nerve in the suboccipital region: a proposal for setting landmarks for local anesthesia in patients with occipital neuralgia. AB - The anatomical relationships of the greater occipital nerve (GON) to the semispinalis capitis muscle (SCM) and the trapezius muscle aponeurosis (TMA) were examined to identify topographic landmarks for use in anesthetic blockade of the GON in occipital neuralgia. The course and the diameter of the GON were studied in 40 cadavers (29 females, 11 males), and the points where it pierced the SCM and the TMA were identified. The course of the GON did not differ between males and females. A left-right difference was detected in the site of the GON in the TMA region but not in the SCM region. The nerve became wider towards the periphery. This may be relevant to entrapment of the nerve in the development of occipital neuralgia. In three cases, the GON split into two branches before piercing the TMA and reunited after having passed the TMA, and it pierced the obliquus capitis inferior muscle in another three cases. The GON and the lesser occipital nerve reunited at the level of the occiput in 80% of the specimens. The occiput and the nuchal midline are useful topographic landmarks to guide anesthetic blockade of the GON for diagnosis and therapy of occipital neuralgia. The infiltration is probably best aimed at the site where the SCM is pierced by the GON. PMID- 16258973 TI - Chiari anomalies in the human right atrium. AB - Chiari anomalies in the human right atrium ostensibly are encountered rarely. There is only sporadic mention in the literature of these fenestrated, net-like valves of the inferior vena cava, coronary sinus, or various strands connecting these with other right atrial structures. The effects of such structural anomalies on heart function are unknown. We report here gross observations of the right atrial net from among 213 cadavers, 38 autopsied, and 11 fetal hearts. Histological and ultrastructural examination of inferior vena cava and coronary sinus valves demonstrated that only the anomalous coronary sinus valves contained cardiac muscle. Chiari anomalies typically have referred to perforations or tissue strands related to the inferior vena cava valve and possibly the coronary sinus valve. The anomaly commonly is cited as occurring in 2% of individuals, although there has been no study to support this. We observed Chiari malformations in 13.6% of the 213 cadaver hearts, and 10.5% of the autopsied hearts examined. Of these malformations, the coronary sinus valve was fenestrated most frequently. We propose the term "right atrial net" for "Chiari net," for anomalies involving valves of the inferior vena cava and coronary sinus, and strands within the right atrium connecting these valves with the crista terminalis, right atrial wall, or interatrial septum. PMID- 16258974 TI - An unusual case of a mediastinal mass in a cadaver. AB - Since 1975, only approximately 60 saphenous vein graft aneurysms have been reported in the medical literature. We now report this unusual finding in a male cadaver who had undergone a recent coronary artery bypass grafting procedure. Although aneurysm formation is an unusual complication of saphenous vein graft surgery, the diagnosis must be suspected particularly in the face of a mediastinal mass on chest radiography in patients who have undergone coronary artery bypass grafting. Needle biopsy in these cases may prove lethal. We believe this to be the first description of this complication in a cadaver. PMID- 16258975 TI - Phase II study of pemetrexed in combination with carboplatin in the first-line treatment of advanced nonsmall cell lung cancer. AB - BACKGROUND: The primary objectives of this study were to determine the efficacy and tolerability of a pemetrexed-carboplatin combination as first-line therapy in patients with advanced nonsmall cell lung cancer. METHODS: Eligibility criteria included Zubrod performance status of 0 or 1, Stage IIIB (malignant effusion) or IV disease, and no prior chemotherapy. Treatment was pemetrexed 500 mg/m2 given intravenously and carboplatin area under the serum concentration-time curve = 6 given intravenously on Day 1 every 3 weeks for six cycles; patients could receive additional cycles at the discretion of the treating physician and patient. All patients received folic acid, vitamin B12, and dexamethasone prophylaxis. RESULTS: Fifty patients (31 men and 19 women) were treated. The median age was 62 years. Ninety-six percent of patients had Stage IV disease, and 88% had a performance status of 1. The median number of cycles was 6; 15 patients received 8 or more cycles. There was Grade 3/4 neutropenia in 11 (22%) and 2 (4%) patients, respectively; Grade 3/4 thrombocytopenia in 1 (2%) and 0 patients, respectively. Three patients (6%) experienced Grade 3 nonhematologic side effects (diarrhea, neutropenic pneumonia, and fatigue). No patients had sensory neuropathy or alopecia >Grade 1. The partial response rate was 24%, median time to progression 5.4 months, 1-year survival 56.0%, and median survival 13.5 months. CONCLUSIONS: This is an active, very well-tolerated regimen. Trials focused on how to integrate this doublet with novel agents are warranted. PMID- 16258976 TI - The molecular genetics of gastroenteropancreatic neuroendocrine tumors. AB - The pathobiology of neuroendocrine tumors (NETs) is hampered by the lack of scientific tools that define their mechanisms of secretion, proliferation, and metastasis; and, currently, there are no accurate means to assess tumor behavior and disease prognosis. Molecular biologic techniques and genetic analysis may facilitate the delineation of the molecular pathology of NETs and provide novel insights into their cellular mechanisms. The current status and recent advances in assessment of the molecular basis of tumorigenesis of gastroenteropancreatic neuroendocrine tumors (GEP-NETs) were reviewed (1981-2004). The objectives of this retrospective study were to provide a cohesive overview of the current state of knowledge and to develop a molecular understanding of these rare tumor entities to facilitate the establishment of therapeutic targets and rational management strategies. Multiple differences in chromosomal aberration patterns were noted between gastrointestinal (GI) neuroendocrine and pancreatic endocrine tumors (PETs). Divergence in gene expression patterns in the development of GI carcinoids and PETs was identified, whereas examination of the PET and GI carcinoid data demonstrated only few areas of overlap in the accumulation of genetic aberrations. These data suggest that the recent World Health Organization classification of GEP-NETs may require updating. In addition, previous assumptions of tumor similarity (pancreatic vs. GI) may be unfounded when they are examined at a molecular level. On the basis of the evolution of genetic information, enteric neuroendocrine lesions (carcinoids) and PETs may need to be classified as two distinct entities rather than grouped together as the single entity "GEP-NETs." PMID- 16258977 TI - Left sided abdominal pain in a patient with situs inversus. PMID- 16258978 TI - Biodistribution characteristics of all-trans retinoic acid incorporated in liposomes and polymeric micelles following intravenous administration. AB - The aim of this study was to investigate the biodistribution characteristics of all-trans retinoic acid (ATRA) incorporated in liposomes and polymeric micelles following intravenous administration. [3H] ATRA were incorporated in distearoylphosphatidylcholine (DSPC)/cholesterol (6:4) liposomes. Two types of block copolymers, poly (ethylene glycol)-b-poly-(aspartic acid) derivatives with benzyl (Bz-75) groups, were synthesized to prepare the polymeric micelles for [(3)H]ATRA incorporation. ATRA were dissolved in mouse serum to analyze their inherent distribution. After intravenous administration, the blood concentration of [3H] ATRA in liposomes and polymeric micelles (Bz-75) was higher than that of inherent [3H]ATRA, suggesting that liposomes and polymeric micelles (Bz-75) control the distribution of ATRA. Pharmacokinetic analysis demonstrated that [3H]ATRA incorporated in polymeric micelles (Bz-75) exhibit the largest AUC(blood) and lowest hepatic clearance of ATRA, suggesting that polymeric micelles (Bz-75) are an effective ATRA carrier system for acute promyelocytic leukemia (APL) therapy. These results have potential implications for the design of ATRA carriers for APL patients. PMID- 16258979 TI - Surgical anatomy of the middle turbinate. AB - Clinicians have encountered many variations of the middle turbinate. Previous descriptions of the middle turbinate were only focused on its size and shape and lacked surgical implications associated with endoscopic sinus surgery. Therefore, the aim of this study was to examine the surgical anatomy of the middle turbinate in hemisected cadaveric heads. The middle turbinates from 101 hemisections of adult Korean cadaveric heads were measured using digital calipers and a protractor. The middle turbinates were then classified according to their shape. The mean distance between the anterior attachment of the middle turbinate and the anterior attachment of the superior turbinate was 18.5 mm. The posterior end of the middle turbinate extended more posteriorly than that of the inferior turbinate in 40% of the cases, while in 26.3% of the cases, the posterior end of the inferior turbinate extended more posteriorly than that of the middle turbinate. The middle turbinate was classified into three types according to the shape of its anterior border. In type 1, the anterior border of the middle turbinate ran directly posteroinferiorly from its attachment to the conchal plate, and was observed in 45.3% of the cases. In type 2, the anterior border of the middle turbinate initially coursed inferiorly from the conchal plate and then turned in a posteroinferior direction. This type was observed in 44.2% of the cases. Type 3 involved 10.5% of the cases where the anterior border bulged anteriorly before it coursed posteroinferiorly. The information provided in this report should assist surgeons when performing partial middle turbinectomies. PMID- 16258980 TI - Rapid loss of striatal VMAT2 binding associated with onset of Lewy body dementia. PMID- 16258981 TI - Application of substrate depletion assay for early prediction of nonlinear pharmacokinetics in drug discovery: assessment of nonlinearity of metoprolol, timolol, and propranolol. AB - The purpose of this study was to investigate the advantages of the substrate depletion assay for evaluating linearity of pharmacokinetics compared with the metabolite formation assay. For propranolol, metoprolol, and nisoldipine with multiple and/or sequential metabolisms, the Michaelis constant (Km) and maximum metabolic intrinsic clearance obtained from the depletion assay using rat and human liver microsomes showed a good correlation with relevant parameters with the formation assay. In vitro kinetics and in vivo pharmacokinetic profiles after oral administration of timolol, metoprolol, and propranolol, were investigated in rats using the depletion assay. The same rank order was found between nonlinearities based on dose-normalized areas under the plasma concentration curve (AUC/Dose) and Km values. Using the kinetic parameters of these compounds, AUC was predicted based on a physiological based pharmacokinetic model incorporated saturable metabolism. The AUCs predicted for propranolol and metoprolol had a good relationship with those observed in the in vivo studies, implying that the depletion assay could be useful for assessing linearity of pharmacokinetics. PMID- 16258982 TI - Capillary zone electrophoresis for the estimation of transdermal iontophoretic mobility. AB - The objective of the study was to investigate the relationship between transdermal iontophoretic flux--specifically, the electromigratory component--and electrophoretic mobility as determined by capillary zone electrophoresis (CZE). First, the steady-state iontophoretic transport rates of a series of dipeptides across porcine skin were determined in vitro. Co-iontophoresis of acetaminophen was used to quantify the respective contributions of electroosmosis (EO) and electromigration (EM). Second, the electrophoretic mobilities of the dipeptides and three other cationic drugs (lidocaine, propranolol, and quinine) were determined, under equivalent experimental conditions, using CZE. Analysis of the transport data using the results of the CZE experiments revealed a linear dependence (r2 > 0.9) between EM flux and electrophoretic mobility. The CZE measurements also provided insight into the charge state of "zwitterionic" dipeptides, H-Glu-epsilon-Lys-OH and H-Tyr-Gln-OH, revealing that these molecules had partial net negative charges under the formulation conditions, accounting for the absence of anodal iontophoretic delivery. The results suggest that CZE might (i) enable identification of ionization states of complex molecules, (ii) serve as a preliminary screen to identify electrically mobile compounds suitable for iontophoretic delivery, and (iii) prove useful for predicting the EM contribution to transdermal iontophoretic flux. PMID- 16258983 TI - Neurenteric cyst: case report and a review of the potential dysembryology. AB - We report a 13-year-old female who presented to an outside emergency room following blunt trauma to the head. As part of her evaluation, an unenhanced CT of the face and radiographs of the cervical spine were performed. She was referred to our clinic with "abnormal" imaging. Radiographs revealed dysmorphic cervical spine and ventral clefting of the vertebral bodies from C7-T2. A subsequent MRI was obtained that demonstrated a large spinal cord mass. MRI demonstrated an unenhanced intramedullary mass of the cervicothoracic spinal cord measuring approximately 1.5 cm in greatest axial diameter. Surgical exploration of her intraspinal mass revealed a neurenteric cyst. Spinal neurenteric cysts are categorized in the spectra of occult spinal dysraphism and most likely arise from incomplete separation of the developing notochord and foregut in the embryo. We discuss potential etiologies for the formation of such cysts. The clinician should consider neurenteric cysts in their differential diagnosis of patients with spinal dysraphism and/or intraspinal masses. PMID- 16258984 TI - Significant differences in the disposition of cyclic prodrugs of opioid peptides in rats and guinea pigs following IV administration. AB - The stabilities of DADLE ([D-Ala2,D-Leu5]-Enk, H-Tyr-D-Ala-Gly-Phe-D-Leu-OH), the capped derivative Ac-DADLE-NH2, and the oxymethyl-coumarinic acid (OMCA)-based cyclic prodrug of DADLE and [D-Ala2,Leu5]-Enk (H-Tyr-D-Ala-Gly-Phe-Leu-OH) were determined at 37 degrees C in rat and guinea pig liver microsomes in the presence and absence of paraoxon, an esterase B inhibitor, and ketoconazole, a CYP3A4 inhibitor. These studies showed that the order of stability in microsomes was: DADLE >> Ac-DADLE-NH2 > OMCA-DADLE = OMCA-[D-Ala2,Leu5]-Enk. While paraoxon produced no significant effect on the stability of the studied compounds in liver microsomes, ketoconazole inhibited the metabolism, suggesting that the capped peptide and the cyclic prodrugs are substrates for cytochrome P450 enzymes. For pharmacokinetic studies, the cyclic prodrugs of DADLE and [D-Ala2,Leu5]-Enk were administered i.v. to rats and guinea pigs. Various biological fluids and tissue (brain, bile, and blood) were collected and analyzed for the free peptide and the prodrugs by high performance liquid chromatography with tandem mass spectrometric detection (LC-MS-MS). These studies showed that the conversion of the cyclic prodrugs to the respective linear peptides (i.e., DADLE and [D-Ala2,Leu5]-Enk) was rapid in rat and guinea pig. In terms of drug elimination, only trace amounts of OMCA-DADLE and OMCA-[D-Ala2,Leu5]-Enk were recovered in guinea pig bile (3.3% and 0.82%, respectively), while significant amounts were recovered in rat bile (38.1% and 51.7%, respectively). Brain uptake of the cyclic prodrugs in guinea pigs compared to previously determined brain uptake of OMCA-DADLE in rats was also significantly different. For OMCA-DADLE, the brain levels of the cyclic prodrug and DADLE in guinea pigs were approximately 80 and 8.5 times greater, respectively, than the levels observed in rat brain. The brain-to-plasma prodrug concentration ratios in guinea pigs (>or= 0.6) were significantly higher than the ratio observed in rats (0.01). These species differences are most likely due to the different substrate specificities of the efflux transporters that facilitate liver clearance of these prodrugs and limit their permeation into the brain. PMID- 16258985 TI - Solution behavior of a novel type 1 interferon, interferon-tau. AB - Interferon-tau (IFN-tau) is a novel cytokine that appears during fetal development of mammals. It is currently being investigated for treatment of viral infections and autoimmune diseases. In order to develop a commercial product, a stable formulation will need to be identified. In this study, the solution behavior of IFN-tau was studied using a variety of biophysical methods. The overall structure of IFN-tau is well defined, with the polypeptide chain folding into a four-helix bundle structure, much like other type 1 interferons. However, its solution behavior has not been characterized. The globular structure has a free energy of unfolding of approximately 4 kcal/mole at room temperature. IFN tau was found to remain monomeric upon increasing the protein concentration, even up to 60 mg/mL. The overall structure of IFN-tau is maintained across a pH range of 2-8, but is significantly altered in the presence of nonaqueous solvents. However, IFN-tau appears to refold efficiently when diluted into an aqueous medium from a nonaqueous solution. This behavior allows the protein to be formulated in low water content formulations suitable for use in capsules. PMID- 16258986 TI - Deamidation of model beta-turn cyclic peptides in the solid state. AB - To investigate the importance of secondary structure on peptide deamidation in the solid state, two cyclic beta-turn peptides and their linear analogs were used as models of Asn residues in structured and unstructured domains, and incorporated into poly(vinyl pyrrolidone) (PVP)-based lyophilized solids. The secondary structure of the model peptides was determined in solution and the solid state using a combination of nuclear magnetic resonance (NMR) spectroscopy, circular dichroism (CD), and Fourier transform infrared (FTIR) spectroscopy. The model beta-turn cyclic peptides were found to be type II beta-turns while the linear analogs were determined to be predominantly unstructured. Quantitatively, the cyclic peptides consisted of approximately 80% beta-turn while the linear analogs contained only 30%-35% beta-turn. To characterize the solid environment, T(g), and moisture content of the solid-state formulations were determined. Accelerated stability studies were conducted in the solid state at 37 degrees C using formulations lyophilized from solutions at pH 8.8 (0.1 M borate buffer). The effect of matrix mobility on solid-state deamidation was investigated by altering the moisture content through variation of relative humidity or the addition of a plasticizer. Cyclic peptides degraded 1.2-8 times slower than the linear analogs under all of the conditions studied. The observed rate constants, however, for all of the peptides decreased dramatically (four orders of magnitude) in the glassy solids. This suggests the greater importance of matrix mobility in solid-state degradation. Molecular dynamics (MD) simulations were also performed to explore the low energy, preferred state of the peptides, and determine the structure around the beta-turn. PMID- 16258987 TI - Comparison of statistical analysis and Bayesian Networks in the evaluation of dissolution performance of BCS Class II model drugs. AB - This project compared the effect of formulation variables on the dissolution performance of model Biopharmaceutics Classification System (BCS) Class II drugs from hard gelatin capsules using statistical analysis and Bayesian networks. The drugs chosen for this study were carbamazepine (CAR), chlorpropamide (CHL), diazepam (DIA), ketoprofen (KET), and naproxen (NAP). Formulations contained anhydrous lactose, microcrystalline cellulose, sodium stearyl fumerate, sodium lauryl sulfate, and croscarmellose sodium. A Box-Behnken experimental design was used in the statistical analysis. The weakly acidic drugs were tested using USP apparatus II with capsule sinkers in 0.1M pH 6.8 Potassium Phosphate buffer. The weakly basic drugs were tested using USP apparatus I in 0.1N HCl buffer. Mean dissolution profiles were compared via calculation of the similarity factor. The Box-Behnken experimental design was found to be useful in assessing primary and secondary excipient effects on dissolution. The Bayesian Network developed for the dataset mirrored the key excipient effects on dissolution performance. PMID- 16258988 TI - Quantitation of crystalline and amorphous forms of anhydrous neotame using 13C CPMAS NMR spectroscopy. AB - Although most drugs are formulated in the crystalline state, amorphous or other crystalline forms are often generated during the formulation process. The presence of other forms can dramatically affect the physical and chemical stability of the drug. The identification and quantitation of different forms of a drug is a significant analytical challenge, especially in a formulated product. The ability of solid-state 13C NMR spectroscopy with cross polarization (CP) and magic-angle spinning (MAS) to quantify the amounts of three of the multiple crystalline and amorphous forms of the artificial sweetener neotame is described. It was possible to quantify, in a mixture of two anhydrous polymorphic forms of neotame, the amount of each polymorph within 1-2%. In mixtures of amorphous and crystalline forms of neotame, the amorphous content could be determined within 5%. It was found that the crystalline standards that were used to prepare the mixtures were not pure crystalline forms, but rather a mixture of crystalline and amorphous forms. The effect of amorphous content in the crystalline standards on the overall quantitation of the two crystalline polymorphic forms is discussed. The importance of differences in relaxation parameters and CP efficiencies on quantifying mixtures of different forms using solid-state NMR spectroscopy is also addressed. PMID- 16258989 TI - Biophysical signatures of noncovalent aggregates formed by a glucagonlike peptide 1 analog: a prototypical example of biopharmaceutical aggregation. AB - LY307161 is a 31 amino acid analog of glucagonlike peptide-1(7-37)OH susceptible to physical instability associated with pharmaceutical processing. Orthogonal biophysical studies were conducted to explore the origins of this physical instability and to distinguish pharmaceutically desirable states of this aggregating peptide from undesirable ones. Equilibrium sedimentation analysis established that LY307161 exists as a monomer at pH 3, and reversibly self associates in the pH range 7.5-10.5. Causative factors for physical instability related to lyophilization conditions were investigated. Solution pH, acetonitrile content, and concentration of the peptide prior to lyophilization each impacted physicochemical properties of the resultant powders. A comparative study of two powder samples exhibiting physicochemically disparate properties established that LY307161 forms soluble noncovalent aggregates. FT-IR analyses in the solid and solution states identified a prominent band at 1657-1659 cm(-1) attributed to alpha-helix structure. Noncovalent soluble aggregate exhibited characteristic bands at 1615 and 1698 cm(-1) indicative of intermolecular beta-sheet structure. An agitation-induced, precipitated solid form of LY307161 exhibited a different FT-IR signature indicative of a conformationally distinct species. Circular dichroism and fluorescence spectroscopy, together with dynamic light scattering measurements and dye-aggregate complexation, provided additional insights into the distinctions between aggregated and native LY307161. PMID- 16258990 TI - Determination of polymorph conversion of an active pharmaceutical ingredient in wet granulation using NIR calibration models generated from the premix blends. AB - Near infrared (NIR) spectroscopy was used for quantitatively monitoring polymorph conversion of an active pharmaceutical ingredient (API) in wet granulation. The API under pharmaceutical development has two different polymorphs. Polymorph A is the stable form for drug development and polymorph B is the undesired form produced at elevated temperature and humidity. Because a reference method was not available for quantitation of polymorph B, a calibration set was not readily available for NIR method development. Analysis of NIR spectra of different polymorphs of the API, the premix blend and wet granulation samples revealed narrow spectral regions, which were unique to polymorph B and insensitive to differences in physical properties between the premix blend and wet granulation. Therefore, the premix blend samples spiked with polymorph B were used as the calibration set. The final univariate NIR method can be used for off-line or on line monitoring of polymorph conversion in the wet granulation process. PMID- 16258991 TI - Formation and characterization of porous indomethacin-PVP coprecipitates prepared using solvent-free supercritical fluid processing. AB - Supercritical carbon dioxide (sc-CO2) was used to prepare coprecipitates of indomethacin (IM) and poly(vinylpyrrolidone) (PVP) with the aim to improve the dissolution rate of IM. The coprecipitates of IM and PVP at various proportions were prepared using a stirred batch reactor containing sc-CO2 as a gas saturated solution (i.e., the compressible CO2 is dissolved in the molten compound). Temperatures between 40 and 90 degrees C and pressure of 150 or 200 bar were employed. The coprecipitates prepared at 75 degrees C and 150 bar were characterized using differential scanning calorimetry (DSC), powder X-ray diffraction (PXD), scanning electron microscopy (SEM), and dissolution testing. The results suggested that IM was totally amorphous at PVP weight fraction of 0.80 and above (indeed, as a molecular composite in which the drug molecules interact with the polymer backbone). As the PVP weight fraction decreased, IM displayed an increasing amount of crystalline material. The SEM photographs of coprecipitates showed a foamed and porous structure. The dissolution rate of IM was increased by incorporation of PVP. IM and PVP at various weight fractions exhibited comparatively higher dissolution rates than that of crystalline IM alone. The sc-CO2 based process produced a solvent free, completely amorphous porous IM solid dispersion with a rapid dissolution rate. PMID- 16258992 TI - pH-dependent functional activity of P-glycoprotein in limiting intestinal absorption of protic drugs: kinetic analysis of quinidine efflux in situ. AB - The purpose of this investigation is to evaluate the quantitative contribution of pH-dependent passive permeability on the functional activity of P-glycoprotein (P gp) in limiting intestinal absorption of weakly basic drugs, in order to include this effect in prediction models. pH-dependent octanol/buffer partition coefficient, artificial membrane permeability and in situ rat intestinal permeability of quinidine were determined in the physiological pH range of gastrointestinal tract. In situ permeability, as a function of luminal pH, was also determined in the presence of P-gp inhibitor, verapamil (500 microM). Octanol/buffer partition coefficient, transport across artificial membrane, and rat in situ permeability showed high pH-dependency. Absorption quotient (AQ), calculated from in situ permeability to express the functional activity of P-gp, declined with increase in luminal pH or increase in luminal quinidine concentration because of the increased passive permeability or saturation of P gp. AQ was 0.57 +/- 0.02 and 0.41 +/- 0.05, while passive permeability was 0.32 +/- 0.01 x 10(-4) cm/sec and 0.43 +/- 0.02 x 10(-4) cm/sec, in jejunum and ileum, respectively, at pH 7.4. Further, apparent Michaelis-Menten constants (K(M), J(P gp,max)) for the quinidine efflux in jejunum indicated that efflux activity was more at luminal pH 4.5 over pH 7.4. K(M) values for jejunum quinidine efflux at pH 4.5 and pH 7.4 were determined to be 77.63 +/- 10.90 and 22.86 +/- 5.22 microM, with J(P-gp,max) values of 1.47 +/- 0.08 and 0.62 +/- 0.04 nM/cm2/sec, respectively. AQ vs passive permeability showed significant relationship indicating dependency of P-gp-mediated efflux on pH-dependent passive permeability, which is dictated by ionization status for a protic or ampholytic drug. In conclusion, an orally administered drug is absorbed from various segments of intestine, which inherit difference in luminal pH, transcellular permeability and P-gp expression. In situ data suggests that pH-dependency and regional variability in passive permeability of protic substrates significantly influence their P-gp-mediated efflux and may have implications on predictions of the in vivo drug absorption. PMID- 16258993 TI - Dose dependency in the oral bioavailability of an organic cation model, tributylmethyl ammonium (TBuMA), in rats: association with the saturation of efflux by the P-gp system on the apical membrane of the intestinal epithelium. AB - The oral bioavailability of tributylmethyl ammonium (TBuMA), an organic cation (OC), exhibited a dose-dependency (i.e., 17, 27, and 35% at doses of 0.4, 4, or 12 micromol/kg, respectively) in the rat. Relevant mechanisms were investigated in the present study by estimating the mucosal to serosal (m-s) and serosal to mucosal (s-m) transport of TBuMA across the rat ileum in an Ussing chamber experiment. The m-s permeability rapidly increased with TBuMA concentration in the mucosal side, and then becoming constant at high TBuMA concentrations. Various studies, including temperature- and potential-dependency and inhibition experiments, revealed that carrier-mediated transport mechanisms (most likely OCT1, OCT3, and P-gp) are involved in the s-m transport of TBuMA, and the saturation of the transport at higher concentrations is responsible for the concentration-dependency in the m-s permeability or dose-dependency of the bioavailability of TBuMA. A nonlinear regression of the m-s transport, based on the assumption of a mixed process of linear diffusion and saturable efflux, exhibited a clearance (CLlinear) of 0.343 microL/min/cm2 for the passive diffusion, and an apparent Km of 241 microM for the saturable process. The Km value is consistent with the concentration range in the intestine which is expected to be achieved after the oral dosing of TBuMA at a dose of 0.4 micromol/kg (i.e., 68 approximately 185 microM). Interestingly, the m-s transport of TBuMA was increased by the presence of P-gp substrates or inhibitors in the mucosal side, but not by the mucosal presence of OCT substrates or inhibitors, suggesting that only efflux transport systems on the apical membrane (e.g., P gp), but not those on the serosal membrane (e.g., OCT1 and OCT3), of the intestinal epithelial cells, are involved in the dose-dependency or concentration dependency. A similar relationship seems likely for drugs that are substrates of efflux transporters on the apical membrane of the intestinal epithelium. PMID- 16258994 TI - Supercritical fluid crystallization of griseofulvin: crystal habit modification with a selective growth inhibitor. AB - Poly (sebacic anhydride) (PSA) was used as a growth inhibitor to selectively modify habit of griseofulvin crystals formed via the Precipitation with a compressed-fluid antisolvent (PCA) process. PSA and griseofulvin were coprecipitated within a PCA injector, which provided efficient mixing between the solution and compressed antisolvent process streams. Griseofulvin crystal habit was modified from acicular to bipyramidal when the mass ratio of PSA/griseofulvin in the solution feed stream was 3 years and specific activity ranging from 237 to 477 U/g (dry) based on benzylpenicillin hydrolysis). The efficiency of the immobilized rPAC(P.rett) was finally evaluated by studying the kinetically controlled synthesis of beta lactam antibiotics (cephalexin) and estimating the synthesis/hydrolysis ratio (S/H), which is a crucial parameter for the feasibility of the process. PMID- 16259001 TI - A computer-aided approach to compare the production economics of fed-batch and perfusion culture under uncertainty. AB - Fed-batch and perfusion culture dominate mammalian cell culture production processes. In this paper, a decision-support tool was employed to evaluate the economic feasibility of both culture modes via a case study based upon the large scale production of monoclonal antibodies. The trade-offs between the relative simplicity but higher start-up costs of fed-batch processes and the high productivity but higher chances of equipment failure of perfusion processes were analysed. Deterministic analysis showed that whilst there was an insignificant difference (3%) between the cost of goods per gram (COG/g) values, the perfusion option benefited from a 42% reduction in capital investment and a 12% higher projected net present value (NPV). When Monte Carlo simulations were used to account for uncertainties in titre and yield, as well as the risks of contamination and filter fouling, the frequency distributions for the output metrics revealed that neither process route offered the best of both NPV or product output. A product output criterion was formulated and the options that met the criterion were compared based on their reward/risk ratio. The perfusion option was no longer feasible as it failed to meet the product output criterion and the fed-batch option had a 100% higher reward/risk ratio. The tool indicated that in this particular case, the probabilities of contamination and fouling in the perfusion option need to be reduced from 10% to 3% for this option to have the higher reward/risk ratio. The case study highlighted the limitations of relying on deterministic analysis alone. PMID- 16259002 TI - Optimal in-vitro expansion of chondroprogenitor cells in monolayer culture. AB - A continuous production of large quantities of chondroprogenitor cells for the manufacture of engineered cartilage tissue products is required. Expansion of the cell population in vitro has become an essential step in the process of tissue engineering of articular cartilage and the optimization of the culture conditions is a fundamental problem that needs to be addressed. The analysis of both seeding density and passage length was considered crucial in the optimization of expansion processes, and their correct selection should be taken as a requisite to establish culture conditions for monolayer systems. The determination of the optimal seeding density and the corresponding passage length for cell expansion in a serial passaging operation was found to be a compromise between growth kinetics and process time. This optimal determination was carried out using a mathematical approach that led to values of 10(4) cell/cm(2) for seeding density and 73 h for passage length. Additional considerations concerning the running cost of the process were introduced. Although the optimal passage length gave the desired expansion factor in a minimum process time, the selection of an alternative value of 120 h was shown to reduce the cost of the expansion process in more than 60%. The optimization approach presented will contribute to the development of feasible large scale expansion operations of chondroprogenitor cells required by the cartilage tissue engineering industry. PMID- 16259003 TI - Copolymer effects on microglia and T cells in the central nervous system of humanized mice. AB - The random amino acid copolymers FYAK and VWAK ameliorate EAE in a humanized mouse model expressing both a human transgenic myelin basic protein (MBP)85-99 specific T cell receptor and HLA-DR2. Here we show that microglia isolated from the central nervous system (CNS) of humanized mice with EAE induced by MBP85-99 and treated with these copolymers had reduced expression of HLA-DR, and thus reduced capacity to present MBP85-99 and activate transgenic T cells. In vitro microglia up-regulated empty HLA-DR2 upon activation with GM-CSF with or without LPS or IFN-gamma, but not with IL-4 or IL-10. Correspondingly, gene chip arrays showed that the CNS of untreated and YFAK-treated mice differentially expressed pro- and anti-inflammatory molecules during MBP85-99-induced EAE. Interestingly, microglia expressed the full-length gammabeta and alphabeta subunits of the tetrameric adaptor protein complexes AP-1 and AP-2 respectively, but after treatment with GM-CSF these complexes were cleaved, as had been found in immature dendritic cells derived from bone marrow. Strikingly, in vivo the perivascular lymphocyte infiltration seen in untreated mice immunized with MBP85-99 was composed of equal numbers of hVbeta2+ MPB85-99-specific transgenic and hVbeta2- endogenous T cells, while the much smaller infiltration seen after treatment with YFAK was composed predominantly of hVbeta2- endogenous T cells. PMID- 16259004 TI - Hypoxia suppresses the production of matrix metalloproteinases and the migration of human monocyte-derived dendritic cells. AB - As most solid tumors are hypoxic, dendritic cells (DC) in solid tumors are also exposed to hypoxia. While many adaptation responses of tumor cells to hypoxia are known, it is yet to be determined how hypoxia affects the functions of DC. To explore the effects of hypoxia on the functions of DC, we compared the expression of surface markers, cytokines, chemokine receptors and matrix metalloproteinases (MMP) of human monocyte-derived DC (hmDC) differentiated under hypoxia to those differentiated under normoxia. Both groups of hmDC expressed similar levels of surface markers and cytokines. However, expression of MMP-9 and membrane type-1 MMP, as well as migrating activity, was significantly suppressed in hmDC differentiated under hypoxia compared with their normoxia counterparts. We also demonstrated that trichostatin A restored the production of MMP-9 in hmDC, under hypoxia. Collectively, our findings show that a hypoxic microenvironment suppresses the production of MMP in hmDC, most probably through the deacetylation of promoter regions of MMP, thus suppressing the migrating activity of hmDC. Our results suggest that the hypoxic microenvironment in solid tumor tissues may suppress the function of DC. PMID- 16259005 TI - The p38 mitogen-activated protein kinase regulates effector functions of primary human CD4 T cells. AB - The role of p38 mitogen-activated protein kinase in primary human T cells is incompletely understood. We analyzed in detail the role of p38 in the regulation of effector functions and differentiation of human CD4 T cells by using a p38 specific inhibitor and a dominant-negative mutant of p38. p38 was found to mediate expression of IL-10 and the Th2 cytokines IL-4, IL-5, and IL-13 in both, primary naive and memory T cells. In contrast, inhibition of p38 activity did not affect expression of the Th1 cytokines IFN-gamma and TNF induced by TCR stimulation, but decreased IL-12-mediated IFN-gamma expression. Cytokine expression from established Th2 effector cells was also regulated by p38, however, the role of p38 was less pronounced compared to primary CD4 T cells. p38 MAPK regulated cytokine gene expression at both, the transcriptional level by activating gene transcription and the post-transcriptional level by stabilizing cytokine mRNA. As a result of the effect of p38 on IL-4 expression, p38 activity modulated differentiation of naive precursor T cells by inducing a shift of the Th1/Th2 balance toward the immuno-modulatory Th2 direction. Together, the data suggest that p38 plays a key role in human Th2 cell immune responses. PMID- 16259006 TI - The CD3 gamma epsilon/delta epsilon signaling module provides normal T cell functions in the absence of the TCR zeta immunoreceptor tyrosine-based activation motifs. AB - T cell receptor (TCR) signal transduction is mediated by the immunoreceptor tyrosine-based activation motifs (ITAM). The ten ITAM in the TCR complex are distributed in two distinct signaling modules termed TCR zetazeta and CD3 gammaepsilon/deltaepsilon. To delineate the specific role of the zeta ITAM in T cell development and TCR signal transmission, we compared the properties of T cells from different TCR zeta-transgenic lines wherein tyrosine-to-phenylalanine substitutions had been introduced in the zeta subunit. These lines lack selected phosphorylated forms of TCR zeta including just p23, both p21 and p23, or all phospho-zeta derivatives. We report herein that the efficiency of positive selection in HY TCR-transgenic female mice was directly related to the number of zeta ITAM in the TCR. In contrast, TCR-mediated signal transmission and T cell proliferative responses following agonist peptide stimulation were similar and independent of the zeta ITAM. Only the duration of MAPK activation was affected by multiple zeta ITAM substitutions. These results strongly suggest that the ITAM in the CD3 gammaepsilon/deltaepsilon module can provide normal TCR signal transmission, with zeta ITAM providing a secondary function facilitating MAPK activation and positive selection. PMID- 16259007 TI - The multigenic structure of the MHC locus contributes to positive selection efficiency: a role for MHC class II gene-specific restriction. AB - The study of T cell positive selection in the thymus has long been focused on the specificity of the MHC-TCR interactions, making use of genetically manipulated mice that display TCR specificities or selecting peptides of limited diversity. However, little is known on the role of the MHC molecules irrespective of the peptide specificity and the implications of MHC multigenic structure in thymic positive selection have not been addressed. Here, we investigated the effect of MHC class II genetic configuration on the positive selection efficiency of naturally generated pre-selection repertoires in the mouse thymus. Analysis of positively selected thymocyte populations in MHC-congenic and -transgenic mice revealed that expression of I-E molecule in the thymic cortex increases positive selection efficiency of CD4 cells by approximately 50%. We show that increments in positive selection attributable to either the I-A and I-E genes are not due to increased MHC class II expression in the thymic cortex and are not affected by the number of MHC alleles. Collectively, our findings imply that MHC class II gene-restricted TCR specificities significantly contribute to positive selection efficiency, introducing the notion that multigenic structure of the MHC locus serves to increase selection of non-overlapping TCR repertoires. PMID- 16259008 TI - Genetic control of thymic development of CD4+CD25+FoxP3+ regulatory T lymphocytes. AB - Among the several mechanisms known to be involved in the establishment and maintenance of immunological tolerance, the activity of CD4+CD25+ regulatory T lymphocytes has recently incited most interest because of its critical role in inhibition of autoimmunity and anti-tumor immunity. Surprisingly, very little is known about potential genetic modulation of intrathymic regulatory T lymphocyte development. We show that distinct proportions of CD4+CD25+FoxP3+ regulatory T cells are found in thymi of common laboratory mouse strains. We demonstrate that distinct levels of phenotypically identical regulatory T cells develop with similar kinetics in the mice studied. Our experimental data on congenic mouse strains indicate that differences are not caused by the distinct MHC haplotypes of the inbred mouse strains. Moreover, the responsible loci act in a thymocyte intrinsic manner, confirming the latter conclusion. We have not found any correlation between thymic and peripheral levels of regulatory T cells, consistent with known homeostatic expansion and/or retraction of the peripheral regulatory T cell pool. Our data indicate that polymorphic genes modulate differentiation of regulatory T cells. Identification of responsible genes may reveal novel clinical targets and still elusive regulatory T cell-specific markers. Importantly, these genes may also modulate susceptibility to autoimmune disease. PMID- 16259009 TI - Destructive potential of the aspartyl protease cathepsin D in MHC class II restricted antigen processing. AB - Whether specific proteases influence MHC class II antigen presentation is still not clearly defined. Cathepsin D, one of the most abundant lysosomal proteases, is thought to be dispensable for MHC class II antigen presentation, yet in vitro digestions of antigen substrates with endosomes/lysosomes from antigen-presenting cells sometimes reveal a dominant role for pepstatin-sensitive aspartyl proteases of which cathepsin D is the major representative. We tested whether the aspartyl protease substrate myoglobin requires cathepsin D activity for presentation to T cells. Surprisingly, in dendritic cells (DC) lacking cathepsin D, presentation of two different myoglobin T cell epitopes was enhanced rather than hindered. This paradox is resolved by the finding that pepstatin-sensitive myoglobin processing activity persists in lysosomes from cathepsin D-null DC and that this reduced activity, most likely due to cathepsin E, is closer to the optimum level required for myoglobin antigen presentation. Our results indicate redundancy among lysosomal aspartyl proteases and show that while processing activities can be productive for MHC class II T cell epitope generation at one level, they can become destructive above an optimal level. PMID- 16259010 TI - ABCB1 transporter discriminates human resting naive B cells from cycling transitional and memory B cells. AB - The exact identification of B cell subsets is instrumental to understand their dynamics under physiological and pathological conditions. Human memory B cells are currently identified according to the expression of CD27, which is absent on naive B cells. We report here that the ATP-binding cassette (ABC)B1 transporter is exclusively present on mature CD27- naive B cells, while it is absent in CD27+ memory B cells and in a heterogeneous subset of CD27- cells that comprise both switch memory and transitional B cells. Thus, ABCB1 activity precisely discriminates naive from transitional and all memory B cells. Using this improved method to discriminate human B cell subsets, and Ki67 staining to identify recently divided cells, we show that in both cord blood and adult peripheral blood, mature naive B cells are quiescent while transitional B cells and memory B cells have a high in vivo turnover. PMID- 16259011 TI - An essential role for Rxr alpha in the development of Th2 responses. AB - A viable hypomorphic allele of mouse retinoid X receptor alpha (Rxralpha) was created by random germline mutagenesis. The mutation (I273N) alters the ligand binding and heterodimerization domain, and causes a 90% decline in ligand inducible transactivation. Homozygotes develop progressive alopecia and dermal cysts, and progressive exaggeration of Th1 and loss of Th2 responses to antigen. Th1 skewing is directly caused by aberrant function of both antigen-presenting cells and naive CD4 T cells; the predominant Th1 response to antigen is attributable to decreased suppression of regulatory T cells in mutant mouse. Dietary depletion of vitamin A in Th2-prone wild-type mice mimics the immune phenotype caused by the mutation. Hence, RXRalpha plays an important post developmental role in the regulation of adaptive immune responses, and provides a plausible link between nutritional environment and the type of adaptive response that results from immunization. PMID- 16259012 TI - Rapid and sensitive determination of phosphorus-containing amino acid herbicides in soil samples by capillary zone electrophoresis with diode laser-induced fluorescence detection. AB - A straightforward and sensitive method has been developed for the analysis of phosphorus-containing amino acid herbicides (glufosinate and aminomethylphosphonic acid, the major metabolite of glyphosate) in soil samples. For this purpose, the analytical features of two indocyanine fluorescent dyes, sulfoindocyanine succinimidyl ester (Cy5) and 1-ethyl-1-[5-(N-succinimidyl oxycarbonyl)pentyl]-3,3,3,3-tetramethyl-indodicarbocyanine chloride, as labeling reagents for the determination of these herbicides by CZE with diode LIF detection were investigated. Practical aspects related to the labeling chemistry and CZE separation showed that the two probes behave similarly, Cy5 being the best choice for the determination of these herbicides on account of its higher sensitivity. The optimum procedure includes a derivatization step of the pesticides at 25 degrees C for 30 min and direct injection to CZE analysis, which is conducted within about 14 min using ACN in the running buffer. The lowest detectable analyte concentration ranged from 0.025 to 0.18 microg/L with a precision of 3.6-5.4%. These results indicate that indocyanine fluorescence dyes are useful as rapid and sensitive labels for the determination of these herbicides when compared with typical fluorescein dyes such as FITC and 5-(4,6 dichloro-s-triazin-2-ylamino) fluorescein, because they provide faster labeling reactions even at room temperature and the excess of reagent practically does not interfere the determination. Finally, the Cy5 method was successfully applied to soil samples without a preliminary clean-up procedure, and the herbicides were measured without any interference from coexisting substances. The recoveries of these compounds in these samples at fortification levels of 100-500 ng/g were 90 93%. PMID- 16259013 TI - Prostaglandin D2 synthase and its post-translational modifications in neurological disorders. AB - Prostaglandin D2 synthase (PGDS) (beta-trace protein) is a highly abundant cerebrospinal fluid (CSF) glycoprotein. A number of studies have been performed to determine the potential value of this protein for the diagnosis of various neurological disorders. The measurement of total PGDS levels in CSF has proved marginally useful for this purpose, but promising results were obtained while investigating changes in the posttranslational modifications (PTM) pattern. Using 2-DE analysis, we previously showed that PGDS is differentially expressed in ante and post mortem CSF samples. In the present study, we examined whether the PGDS isoforms may help to distinguish stroke and neurodegenerative disease patients from healthy subjects. The pattern of PGDS PTM was analyzed in CSF from patients with various neurological disorders (n = 44) using IEF/immunoblotting techniques. Strong alterations of this pattern were detected in patients with different forms of degenerative dementia. These findings are consistent with PGDS being altered in some neurological diseases and provide new opportunities for clinical applications. PMID- 16259014 TI - Online concentration by field-amplified sample injection in acidic buffer for analysis of fangchinoline and tetrandrine in herbal medicine by flow injection micellar electrokinetic capillary chromatography. AB - A novel, rapid, and continuous online concentration approach based on field amplified sample injection for the analysis of fangchinoline and tetrandrine was developed in this paper by combination of flow injection-MEKC. The BGE used was a solution composed of 75 mM H3PO4-triethylamine-2.5% v/v polyoxyethylene sorbitan monolaurate-20% v/v methanol buffer (pH* 5.0). The analytes prepared in 50% v/v aqueous ethanol were used as the test analytes. Sample was injected electrokinetically between plugs of water. When the cations reached the boundary between the water plug and BGE, they slowed down and became concentrated. Thereafter, MEKC was initiated for the separation. This results in 6.8-8.9-fold improvement in concentration sensitivity relative to conventional CE methods. The separation could be achieved within 10 min and sample throughput rate can reach up to 50/h. The repeatability (defined as RSD) was 4.8, 4.4% with peak height evaluation and 3.6, 0.94% with peak area evaluation for TET and FAN, respectively. PMID- 16259015 TI - Differential staining of Western blots of human secreted glycoproteins from serum, milk, saliva, and seminal fluid using lectins displaying diverse sugar specificities. AB - Human milk, serum, saliva, and seminal fluid glycoproteins (gps) nourish and protect newborn and adult tissues. Their saccharides, which resemble cell membrane components, may block pathogen adhesion and infection. In the present study, they were examined by a battery of lectins from plants, animals, and bacteria, using hemagglutination inhibition and Western blot analyses. The lectins included galactophilic ones from Aplysia gonad, Erythrina corallodendron, Maclura pomifera (MPL), peanut, and Pseudomonas aeruginosa (PA-IL); fucose binding lectins from Pseudomonas aeruginosa (PA-IIL), Ralstonia solanacearum (RSL), and Ulex europaeus (UEA-I), and mannose/glucose-binding Con A. The results demonstrated the chosen lectin efficiency for differential analysis of human secreted gps as compared to CBB staining. They unveiled the diversity of these body fluid gp glycans (those of the milk and seminal fluid being highest): the milk gps interacted most strongly with PA-IIL, followed by RSL; the saliva gps with RSL, followed by PA-IIL and MPL; the serum gps with Con A and MPL, followed by PA-IIL and RSL, and the seminal plasma gps with RSL and MPL, followed by UEA-I and PA-IIL. The potential usage of these lectins as probes for scientific, industrial, and medical purposes, and for quality control of the desired gps is clearly indicated. PMID- 16259016 TI - Nonaqueous capillary electrophoretic assays of p-phenylene-bis-4,4'-(1-aryl-2,6 diphenylpyridinium) molecular wires. AB - A nonaqueous CE system for separation and detection of novel electron-dopable molecular wires based on p-phenylene-bis-4,4'-(1-aryl-2,6-diphenylpyridinium) oligomers is described. The method is based on the coupling of CE separation in pure organic solvent, DMF with 50 mM acetic acid, and UV detection at 338 nm. The system offers a rapid measurement in less than 20 min for two priority nanowires and their impurities. Calibration data confirmed linear response for all compounds of interest in the concentration range 0.1-1.0 mg/mL. A highly stable response was observed for repetitive injections (RSD < or = 2.8%, n = 10). PMID- 16259017 TI - The origin of stereoselectivity in primary amino acid catalyzed intermolecular aldol reactions. PMID- 16259018 TI - Mass spectrometric profiling of intact biological tissue by using desorption electrospray ionization. PMID- 16259021 TI - The catalysis gold rush: new claims. PMID- 16259020 TI - Wrapping peptide tubes: merging biological self-assembly and polymer synthesis. PMID- 16259022 TI - The metal flux: a preparative tool for the exploration of intermetallic compounds. AB - This review highlights the use and great potential of liquid metals as exotic and powerful solvents (i.e. fluxes) for the synthesis of intermetallic phases. The results presented demonstrate that considerable advances in the discovery of novel and complex phases are achievable utilizing molten metals as solvents. A wide cross-section of examples of flux-grown intermetallic phases and related solids are discussed and a brief history of the origins of flux chemistry is given. The most commonly used metal fluxes are surveyed and where possible, the underlying principal reasons that make the flux reaction work are discussed. PMID- 16259025 TI - Sigma-pi energy separation in homodesmotic reactions. AB - A well-established quantity for specifying the aromaticity or antiaromaticity of cyclic conjugated molecules is the so-called aromatic stabilization energy (ASE), which can be derived-either experimentally or theoretically-from appropriate homodesmotic reactions. To gain further insight into the origin of aromaticity, several schemes have been devised to partition ASE into nuclear and electronic as well as sigma and pi contributions, some of which have resulted in contradictory statements about the driving force of aromatic stabilization. Currently, these contradictions have not been resolved and have resulted in a confusing distinction between two different types of aromaticity: extrinsic and intrinsic aromaticity. By investigating different homodesmotic reactions we show that, in contrast to ASE itself, the individual contributions that enter the ASE can strongly depend on the type of reaction. Caution is therefore advised if conclusions or physical interpretations are derived from the individual components. The contradictions result from the fact that some reactions suffer from an imbalance in the number of interaction terms at the two sides of the reaction equation. The concept of isointeractional reactions is introduced and results in the elimination of the imbalance. For these reactions, the contradictions disappear and the distinction between intrinsic and extrinsic aromaticity becomes unnecessary. As far as the sigma-pi partitioning is concerned, several schemes proposed in the literature are compared. Contradictory results are obtained depending on the partitioning scheme and reaction used. In this context, it is demonstrated that for the partitioning of the electron electron interaction, the scheme introduced by Jug and Koster is the one that is most theoretically grounded. PMID- 16259023 TI - Synthesis, structure, and chemoselective reactivity of N-(2 iodylphenyl)acylamides: hypervalent iodine reagents bearing a pseudo-six-membered ring scaffold. PMID- 16259026 TI - In situ 1H NMR study on the trioctylphosphine oxide capping of colloidal InP nanocrystals. AB - We used trioctylphosphine oxide (TOPO) capped colloidal InP nanocrystals (Q InP|TOPO) to explore the potential of solution 1H NMR spectroscopy in studying in situ the capping and capping exchange of sterically stabilized colloidal nanocrystals. The spectrum of Q-InP|TOPO shows resonances of free TOPO, superimposed on broadened spectral features. The latter were assigned to TOPO adsorbed at Q-InP by means of pulsed field gradient diffusion NMR and 1H-13C HSQC spectroscopy. The diffusion coefficient of Q-InP|TOPO nanocrystals was inferred from the decay of the adsorbed TOPO NMR signal. The corresponding hydrodynamic diameter correlates well with the diameter of Q-InP. By using the resolved methyl resonance of adsorbed TOPO, the packing density of TOPO at the InP surface can be estimated. Spectral hole burning was used to demonstrate explicitly that the adsorbed TOPO resonances are heterogeneously broadened. Exchange of the TOPO capping by pyridine was demonstrated by the disappearance of the resonances for adsorbed TOPO and the appearance of pyridine resonances in the 1H NMR spectrum. These results show that solution NMR spectroscopy should be considered a powerful technique for the in situ study of the capping of sterically stabilized colloidal nanocrystals. PMID- 16259027 TI - Model for the conductivity of ionic liquids based on an infinite dilution of holes. PMID- 16259028 TI - Insights into the role of the liquid-liquid interface in biphasic reactions: the reaction of vitamin B12s(aq) with vicinal dibromides(oil). AB - Electrocatalytic processes can take place either homogeneously in a single liquid phase or heterogeneously at the liquid-liquid interface formed in emulsions. This Article addresses the question as to whether a change in rate and/or mechanism can occur between the two possibilities. Specifically, cyclic voltammetry and electrosynthetic experiments are used to demonstrate that for the vitamin B12 mediated reduction of vicinal dibromides producing olefins, electric field effects likely operate at the liquid-liquid interface which can change the populations of different conformers relative to the single homogeneous-phase experiment, leading to significant changes in rate. PMID- 16259029 TI - "Hydrogen-catalyzed" dehydrogenation: a supercritical conundrum. PMID- 16259030 TI - On the chemistry of gold in silicate glasses: studies on a nonthermally activated growth of gold nanoparticles. PMID- 16259031 TI - A triazadiphosphole. PMID- 16259032 TI - Acyl 1,3-migration in rhodium-catalyzed reactions of acetylenic beta-ketoesters with aryl boronic acids: application to two-carbon-atom ring expansions. PMID- 16259033 TI - Squeezing C-C bonds. PMID- 16259035 TI - Controlled hydrothermal synthesis and structural characterization of a nickel selenide series. AB - A series of nickel selenides (NiSe2 microcrystals, Ni(1-x)Se and Ni3Se2 microspheres) has been successfully synthesized through a convenient, low temperature hydrothermal method. A good nucleation and growth environment has been created by forming a uniform and transparent solution reaction system. The compositions (including the x value of Ni(1-x)Se), phase structures, as well as the morphologies of nickel selenides, can be controlled by adjusting the Ni/Se ratio of the raw materials, the pH, the reaction temperatures and times, and so forth. The newly produced Se microspheres in the system have been used as both reactant and in situ template to the Ni(1-x)Se microspheres. It is found that Ni(1-x)Se microspheres act as the intermediate precursor during the formation of Ni3Se2 microspheres. Under certain conditions, hexagonal NiSe microspheres can be converted into rhombohedral NiSe nanowires in solution. The formation mechanisms of a series of nickel selenides has been investigated in detail by means of X-ray diffraction (XRD) and scanning electron microscopy (SEM) analyses. This work has provided a general, simple, and effective method to control the composition, phase structure, and morphology of metal selenides in aqueous solution, which will be important for inorganic synthesis methodology and further applications of selenides. PMID- 16259034 TI - Numerical simulation of the effect of solvent viscosity on the motions of a beta peptide heptamer. AB - This report examines the effect of a decrease in solvent viscosity on the simulated folding behaviour of a beta-peptide heptamer in methanol. Simulations of the molecular dynamics of the heptamer H-beta3-HVal-beta3-HAla-beta3-HLeu (S,S)-beta3-HAla(alphaMe)-beta3-HVal-beta3-HAla-beta3-HLeu-OH in methanol, with an explicit representation of the methanol molecules, were performed for 80 ns at various solvent viscosities. The simulations indicate that at a solvent viscosity of one third of that of methanol, only the dynamic aspects of the folding process are altered, and that the rate of folding is increased. At a viscosity of one tenth of that of methanol, insufficient statistics are obtained within the 80 ns period. We suggest that 80 ns is an insufficient time to reach conformational equilibrium at very low viscosity because the dependence of the folding rate of a beta-peptide on solvent viscosity has two regimes; a result that was observed in another computational study for alpha-peptides. PMID- 16259036 TI - DFT-HSAB prediction of regioselectivity in 1,3-dipolar cycloadditions: behavior of (4-substituted)benzonitrile oxides towards methyl propiolate. AB - The regioselectivity of 1,3-dipolar cycloadditions between (4 substituted)benzonitrile oxides and methyl propiolate cannot be rationalized on the basis of the electron demand of the reactants or frontier molecular-orbital theory. To this problem, we have applied a quantitative formulation of the hard soft acid-base principle developed within the density functional theory. Global and local reactivity indices were computed at B3LYP/6-311+G(d,p) level. The details of charge transfer upon the reactive encounter have been elucidated, and the computed regioselectivity has been shown to be in good agreement with experimental data. PMID- 16259038 TI - Assignment of absolute configuration of cyclic secondary amines by NMR techniques using Mosher's method: a general procedure exemplified with (-)-isoanabasine. AB - A general procedure to determine the absolute configuration of cyclic secondary amines with Mosher's NMR method is demonstrated, with assignment of absolute configuration of isoanabasine as an example. Each Mosher amide can adopt two stable conformations (named rotamers) caused by hindered rotation around amide C- N bond. Via a three-step structural analysis of four rotamers, the absolute configuration of (-)-isoanabasine is deduced to be (R) on the basis of Newman projections, which makes it easy to understand and clarify the application of Mosher's method to cyclic secondary amines. Furthermore, it was observed that there was an unexpected ratio of rotamers of Mosher amide derived from (R) isoanabasine and (R)-Mosher acid. This phenomenon implied that it is necessary to distinguish the predominant rotamer from the minor one prior to determining the absolute configuration while using this technique. PMID- 16259039 TI - (13)C NMR spectral assignment of 1,4-diarylpiperazinones. AB - The piperazinone derivatives have potential application in the pharmaceutical, polymer and textile fields. The present work describes the preparation of a series of new 1,4-diarylsubstituted-2-piperazinones by condensation of substituted N,N'-bis-(2-hydroxyphenyl)-ethylenediamines with glyoxal and the complete (13)C NMR spectral assignment accomplished using APT, HMQC and HMBC techniques. Substituent chemical-shift effects (SCS) were calculated, which showed different values for the lactam- and amine-substituted aromatic rings. The results show that predictions based on SCS effects are not simple for these molecules due to electronic and steric effects. Moreover, in the case of the ortho-substituted derivative 2 g, the NMR spectra reveal a dynamic behavior related to restricted rotation of the phenyl groups (atropisomerism). PMID- 16259041 TI - Laser ablation synthesis of selenium superoxide anion SeO4- via selenium trioxide photolysis. Time-of-flight mass spectrometry and ab initio calculations. AB - Laser desorption/ionisation and laser ablation of solid selenium trioxide, as well as the gas-phase behaviour of selenium trioxide, were studied. Selenium trioxide undergoes photochemical decomposition and, from the mass spectra obtained by laser desorption/ionisation time-of-flight mass spectrometry (LDI-TOF MS), the following species were identified: O-, O2-, O3-, SeO-, SeO2-, SeO3-, SeO4-, Se2O7-, Se3O11-, and Se4O14-. Formation of the selenium superoxide SeO4- anion is described in this work for the first time. In addition, low-abundance selenium species such as Se2O8H2-, Se3O11H-, and Se4O15H2- were also detected. The stoichiometry of all ions was confirmed via isotopic pattern modeling and/or post-source decay (PSD) analysis. Photolysis of selenium trioxide leads partly to ozone formation. It was found that the most likely mechanisms of selenium superoxide formation are oxidation of selenium trioxide with ozone and/or reactive oxygen radicals, or photolysis of selenium trioxide tetramer (SeO3)4. Therefore, ab initio calculations were performed to support the mass spectrometric evidence and to suggest probable geometries for selenium superoxide anion SeO4- and diselenium superoxide anion Se2O7-, as well as to provide insight into and/or predict possible formation pathways. It has been found that both cyclic and non-cyclic peroxide structures of SeO4- and Se2O7- ions are possible. In addition, the SeO4 structure was also calculated guided by thermodynamic considerations using Gaussian-2 methodology, and the inferred stability of the SeO4 neutral molecule was supported by ab initio calculations. PMID- 16259040 TI - Post-translational modifications of recombinant B. cinerea EPG 6. AB - The fungus Botrytis cinerea is a ubiquitous plant pathogen that infects more than 200 different plant species and causes substantial economic losses in a wide range of agricultural crops and harvested products. Endopolygalacturonases (EPGs) are among the first array of cell-wall-degrading enzymes secreted by fungi during infection. Up to 13 EPG glycoforms have been described for B. cinerea. The presence of multiple N-linked glycosylation modifications in BcPG1-6 is predicted by their deduced amino acid sequences. In this work, the glycosylation sites and the attached oligosaccharide structures on BcPG6 were analyzed. The molecular mass of the intact glycoprotein was determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometric (MALDI-TOFMS) analysis. BcPG6 contains seven potential N-linked glycosylation sites. Occupancy of these glycosylation sites and the attached carbohydrate structures were analyzed by tryptic digestion followed by liquid chromatography/mass spectrometry (LC/MS) using a stepped orifice voltage approach. Five out of seven potential N-linked sites present in BcPG6 were determined to be occupied by high-mannose-type oligosaccharides. Four of them were readily determined to be at Asn58 (T3 peptide), Asn198 (T7 peptide), Asn237 (T9 peptide) and Asn256 (T11 peptide), respectively. Another was located on the T8 peptide, which contained two potential N-linked sites, Asn224 and Asn227 (SNNN224VTN227ITFK). LC/MS/MS of a sample treated with N-glycanase placed the glycan in this peptide at Asn224 rather than at Asn227. The potential glycosylation site on Asn146 (T6 peptide) was not glycosylated. In addition, two disulfide bonds were observed, linking the Cys residues within the T13 and T16 peptides. PMID- 16259042 TI - Determination of arecoline (areca nut alkaloid) and nicotine in hair by high performance liquid chromatography/electrospray quadrupole mass spectrometry. PMID- 16259043 TI - Cu(II)-loaded iminodiacetic acid-silica particles for protein profiling of human serum samples using surface-enhanced affinity capture: support porosity effects. AB - Silica particles of different porosity were functionalised with iminodiacetic acid (IDA) and loaded with Cu(II) ions to yield Cu(II)-IDA-silica. These immobilised metal affinity chromatography (IMAC) materials were subjected to a comprehensive characterisation study. The Cu(II) content--determined via UV/Vis spectroscopy and atomic absorption spectroscopy (AAS)--was found to be linearly dependent on the specific surface area of the silica particles. The evaluation of protein adsorption isotherms provided information on binding properties towards biomolecules. The data fitted Langmuir's adsorption theory. Binding capacity of hen egg white lysozyme (HEWL) was highest for Cu(II)-IDA-silica with mean pore diameter of 120 A, reaching nearly 350 mg/g. All derivatised materials were applied to the fractionation of human serum samples and subsequent mass spectrometric analysis (m/z: 2000-10,000) according to a surface-enhanced affinity capture (SEAC) protocol. Pore size of the support material affected the appearance of the mass spectra to a great extent, showing that surface morphology is another parameter that has to be considered in addition to surface chemistry. Signal intensity as well as the number of detected masses were strongly dependent on the pore diameter, indicating that the carrier material has to be carefully chosen to assure best results. PMID- 16259044 TI - Mass spectrometric analysis of the interactions between CP12, a chloroplast protein, and metal ions: a possible regulatory role within a PRK/GAPDH/CP12 complex. AB - The small chloroplast protein CP12 plays the role of a protein linker in the assembly process of a PRK/GAPDH/CP12 complex that is involved in CO2 assimilation in photosynthetic organisms. The redox state of CP12 regulates its role as a protein linker. Only the oxidized protein, with two disulfide bonds, is active in complex formation. Several observations indicating that CP12 might bind a metal ion led us to screen the binding of different metal ions on oxidized or reduced CP12 using non-covalent electrospray ionization mass spectrometry (ESI-MS) experiments. The oxidized protein bound specifically Cu2+ and Ni2+ (Kd of 26+/-1 microM and 11+/-1 microM, respectively); other cations such as Fe2+ and Zn2+ did not bind, while cations such as Cd2+ formed non-specific adducts to CP12. Similar results were obtained for metal ions on screening with the reduced CP12. Interestingly, the present results suggest that Cu2+ catalyzes the re-formation of the disulfide bonds of the reduced CP12, leading to recovery of the fully oxidized CP12 that is then able to bind a Cu2+ ion. Finally the high similarity between CP12 and copper chaperones from Arabidopsis thaliana, as judged by hydrophobic cluster analysis, provides additional evidence for the relevance of metal binding for the in vivo situation. The findings that CP12 is able to bind a metal ion, and that Cu2+ catalyzes the oxidation of the thiol groups of CP12, are new characteristics of this protein that may prove to be important in the regulation of the assembly process of the PRK/GAPDH/CP12 complex. PMID- 16259045 TI - Specific detection of Lewis x-carbohydrates in biological samples using liquid chromatography/multiple-stage tandem mass spectrometry. AB - The Lewis x structure [Lex, Galbeta1-4(Fucalpha1-3)GlcNAc] motif is one of the tumor antigens and plays an important role in oncogenesis, development, cellular differentiation and adhesion. The detection of Lex-carbohydrates and their structural analysis are necessary to clarify the role of Lex in several biological events. Mass spectrometry has been preferably used for the structural analysis of carbohydrates. Especially, collision-induced dissociation (CID) tandem mass spectrometry (MS/MS), which causes a glycosidic bond cleavage, is used for carbohydrate sequencing. However, Lex cannot be identified by MS/MS due to the existence of the positional isomers, such as Lewis a [Galbeta1-3(alpha1 4Fuc)GlcNAc]. In the present study, we demonstrate the specific detection of Lex carbohydrates in a biological sample by using multiple-stage MS/MS (MSn). Using pyridylaminated oligosaccharides bearing Lex, we found that the Lex-motif yields a cross-ring fragment by the cleavage of a bond between C-3 and C-4 of GlcNAc in Gal(Fuc)GlcNAc. The Lex-specific cross-ring fragment ion at m/z 259 was effectively detected by sequential scans, consisting of a full MS1 scan, data dependent CID MS2 scan, MS3 of [Gal(Fuc)GlcNAc+Na]+ at m/z 534, and MS4 of [GalGlcNAc+Na]+ at m/z 388. The sequential scan was applied to N-linked oligosaccharide profiling using a LC/ESI-MSn system equipped with a graphitized carbon column. We successfully detected the Lex-motif and elucidated the structures of several Lex and Lewis y [(Fucalpha1-2)Galbeta1-4(Fucalpha1 3)GlcNAc] oligosaccharides in the murine kidney used as a model tissue. Our method is expected to be a powerful tool for the specific detection of the Lex motif, and structural elucidation of Lex-carbohydrates in biological samples. PMID- 16259046 TI - Mass spectrometry of stanozolol and its analogues using electrospray ionization and collision-induced dissociation with quadrupole-linear ion trap and linear ion trap-orbitrap hybrid mass analyzers. AB - Mass spectrometric identification and characterization of growth-promoting anabolic-androgenic steroids in biological matrices has been a major task for doping control as well as food safety laboratories. The fragmentation behavior of stanozolol, its metabolites 17-epistanozolol, 3'-OH-stanozolol, 4alpha-OH stanozolol, 4beta-OH-stanozolol, 17-epi-16alpha-OH-stanozolol, 16alpha-OH stanozolol, 16beta-OH-stanozolol, as well as the synthetic analogues 4 dehydrostanozolol, 17-ketostanozolol, and N-methyl-3'-OH-stanozolol, was investigated after positive electrospray ionization and subsequent collision induced dissociation utilizing a quadrupole-linear ion trap and a novel linear ion trap-orbitrap hybrid mass spectrometer. Stable isotope labeling, H/D-exchange experiments, MS3 analyses and high-resolution/high mass accuracy measurements of fragment ions were employed to allow proposals for charge-driven as well as charge-remote fragmentation pathways generating characteristic product ions of stanozolol at m/z 81, 91, 95, 105, 119, 135 and 297 and 4-hydroxylated stanozolol at m/z 145. Fragment ions were generated by dissociation of the steroidal A- and B-ring retaining the introduced charge within the pyrazole function of stanozolol and by elimination of A- and B-ring fractions including the pyrazole residue. In addition, a charge-remote fragmentation causing the neutral loss of methanol was observed, which was suggested to be composed by the methyl residue at C-18 and the hydroxyl function located at C-17. PMID- 16259047 TI - Determination of microbial volatile organic compounds from Staphylococcus pasteuri against Tuber borchii using solid-phase microextraction and gas chromatography/ion trap mass spectrometry. AB - The mycelium of Tuber borchii Vittad., a commercial truffle species, is used as a model system for in vitro ectomycorrhizal synthesis, infected seedling production and biotechnological applications. Our fungal cultures were accidentally contaminated with a Staphylococcus pasteuri strain, showing a strong antifungal activity against T. borchii mycelium. In order to identify the antifungal volatile agents produced by S. pasteuri, solid-phase microextraction (SPME) with gas chromatography and mass spectrometry (GC/MS) was used. Using this method 65 microbial volatile organic compounds (MVOCs), synthesized by this bacterium in either single or in fungal-bacterial dual culture, were identified. SPME combined with GC/MS may be a useful method for the determination of MVOCs involved in the antifungal activity. These results showed that bacteria with unusual biological activities could be a major problem during large-scale production of inoculum for truffle-infected seedling. PMID- 16259048 TI - Bayesian cost-effectiveness analysis with two measures of effectiveness: the cost effectiveness acceptability plane. AB - Cost-effectiveness analysis (CEA) compares the costs and outcomes of two or more technologies. However, there is no consensus about which measure of effectiveness should be used in each analysis. Clinical researchers have to select an appropriate outcome for their purpose, and this choice can have dramatic consequences on the conclusions of their analysis. In this paper we present a Bayesian cost-effectiveness framework to carry out CEA when more than one measure is considered. In particular, we analyse the case in which two measures of effectiveness, one binary and the other continuous, are considered. Decision making measures, such as the incremental cost-effectiveness ratio, incremental net-benefit and cost-effectiveness acceptability curves, are used to compare costs and one measure of outcome. We propose an extension of cost-acceptability curves, namely the cost-effectiveness acceptability plane, as a suitable measure for decision taking. The models were validated using data from two clinical trials. In the first one, we compared four highly active antiretroviral treatments applied to asymptomatic HIV patients. As measures of effectiveness, we considered the percentage of patients with undetectable levels of viral load, and changes in quality of life, measured according to EuroQol. In the second clinical trial we compared three methadone maintenance programmes for opioid-addicted patients. In this case, the measures of effectiveness considered were quality of life, according to the Nottingham Health Profile, and adherence to the treatment, measured as the percentage of patients who participated in the whole treatment programme. PMID- 16259050 TI - Eremophilane esters of Robinsonecio gerberifolius and their rearranged products. Study of the coupling constants (2)J(H, H), (3)J(H, H) and (4)J(H, H). AB - In the course of the basic hydrolysis of four eremophilane esters isolated from Robinsonecio gerberifolius, some rearrangements, eliminations, and additions occurred. Five compounds were obtained, three of them not previously described. Additionally, a new sesquiterpene was produced by autooxidation of compound 1. The (1)H and (13)C NMR spectra of these compounds were completely assigned by utilization of HMQC, HMBC, COSY, DEPT, and NOESY techniques. The long-range coupling constants of the peroxide 10 are reported, and all its coupling constants (2)J(H, H), (3)J(H, H), and (4)J(H, H) are calculated at the B3LYP/6 31G(d,p) level of theory. Their magnitude is explained in terms of electronic delocalization and the additivity of stereoelectronic effects. PMID- 16259049 TI - Sometimes more equal than others: how health inequalities depend on the choice of welfare indicator. AB - In recent years, a large body of empirical work has focused on measuring and explaining socio-economic inequalities in health outcomes and health service use. In any effort to address these questions, analysts must confront the issue of how to measure socioeconomic status. In developing countries, socioeconomic status has typically been measured by per capita consumption or an asset index. Currently, there is only limited information on how the choice of welfare indicators affect the analysis of health inequalities and the incidence of public spending. The purpose of this paper is to illustrate the potential sensitivity of the analysis of health related inequalities to how socioeconomic status is measured. Using data from Mozambique, the paper focuses on five key health service indicators, and tests whether measured inequality (concentration index) in health service utilization differs depending on the choice of welfare indicator. The paper shows that, at least in some contexts, the choice of welfare indicator can have a large and significant impact on measured inequality in utilization of health services. In consequence, we can reach very different conclusions about the 'same' issue depending on how we define socioeconomic status. The paper also provides some tentative conclusions about why and in what contexts health inequalities can be sensitive to the choice of living standards measure. The results call for more clarity and care in the analysis of health related inequalities, and for explicit recognition of the potential sensitivity of findings to the choice of welfare measure. The results also point at the need for more careful research on how different dimensions of SES are related, and on the pathways by which the respective different dimensions impact on health related variables. PMID- 16259052 TI - Carbon-13 and proton NMR assignments of a new agathisflavone derivative. AB - The (1)H and (13)C NMR spectra of 5-acetyl-7,4'-dimethoxyflavone-(6-8'')-5'' acetyl-7'',4'''-dimethoxyflavone, a new agathisflavone derivative, were completely assigned on the basis of 1D and 2D NMR techniques. PMID- 16259051 TI - Dicondensed indolinobenzospiropyrans as precursors of thermo- and photochromic spiropyrans. Part II: Assignment of (1)H and (13)C NMR spectra. AB - The (1)H and (13)C NMR spectra of dicondensed indolinobenzospiropyrans as precursors of thermo- and photochromic spiropyrans, DC1-DC5, were completely assigned. Especially, the (1)H assignment and coupling characteristics of the diastereotopic protons at the carbon-3 position of the benzopyran rings were achieved by conducting (1)H-(1)H COSY and nOe experiments. The dihedral angles (theta(1), theta(2) and theta(3)) calculated from the experimental values of the vicinal coupling constants ((3)J) of DC5 are in good agreement with the observed values in the solid state. All of the carbons in the DC dye molecules were investigated through a combination of heteronuclear 2D-shift correlation spectroscopy (HETCOR) and DEPT135. PMID- 16259053 TI - Complete assignment of (1)H and (13)C NMR data of dihydroxyflavone derivatives. AB - Six flavone derivatives were studied. Previously reported NMR data of three of these derivatives were corrected and the NMR data for the other three derivatives not studied previously were completely assigned on the basis of the basic 1D and 2D NMR experiments and molecular modeling. PMID- 16259054 TI - Different types of hydrogen bonds in 2-substituted pyrroles and 1-vinyl pyrroles as monitored by (1)H, (13)C and (15)N NMR spectroscopy and ab initio calculations. AB - According to the (1)H, (13)C and (15)N NMR spectroscopic data and ab initio calculations, the strong N--H...O intramolecular hydrogen bond in the Z-isomers of 2-(2-acylethenyl)pyrroles causes the decrease in the absolute size of the (1)J(N,H) coupling constant by 2 Hz in CDCl(3) and by 4.5 Hz in DMSO-d(6), the deshielding of the proton and nitrogen by 5-6 and 15 ppm, respectively, and the lengthening of the N--H link by 0.025 A. The N--H...N intramolecular hydrogen bond in the 2(2'-pyridyl)pyrrole leads to the increase of the (1)J(N,H) coupling constant by 3 Hz, the deshielding of the proton by 1.5 ppm and the lengthening of the N--H link by 0.004 A. The C--H...N intramolecular hydrogen bond in the 1 vinyl-2-(2'-pyridyl)-pyrrole results in the increase of the (1)J(C,H) coupling constant by 5 Hz, the deshielding of the proton by 1 ppm and the shortening of the C--H link by 0.003 A. Different behavior of the coupling constants and length of the covalent links under the hydrogen bond influence originate from the different nature of the hydrogen bonding (predominantly covalent or electrostatic), which depends in turn on the geometry of the hydrogen bridge. The Fermi-contact mechanism only is responsible for the increase of the coupling constant in the case of the predominantly electrostatic hydrogen bonding, whereas both Fermi-contact and paramagnetic spin-orbital mechanisms bring about the decrease of coupling constant in the case of the predominantly covalent hydrogen bonding. PMID- 16259057 TI - Development of the somatic stress response scale and its application in clinical practice. AB - The objective of this study was to develop the Somatic Stress Response Scale (SSRS), and then to use the scale in clinical practice. A preliminary survey was conducted using 109 healthy adults to obtain somatic stress responses. Then, 215 healthy subjects completed a preliminary questionnaire. A comparison was made regarding the somatic stress responses among 191 patients (71 with anxiety disorders, 73 with depressive disorders and 47 with somatoform disorders) and 215 healthy subjects. Factor analysis of the SSRS yielded five subscales: the cardiorespiratory response, somatic sensitivity, gastrointestinal response, general somatic response and genitourinary response subscales. The test-retest reliability for the five subscales and the total score was significantly high, ranging from .86 to .94. The Cronbach's afor the five subscales ranged from .72 to .92, and was .95 for the total score. By correlating the five subscales and the total score of the SSRS with the somatization subscale scores of the Symptom Checklist-90-Revised (SCL-90-R), convergent validity was calculated. The correlations were all at significant levels. Each of the disorder groups was significantly higher in scores of the cardiorespiratory response, gastrointestinal response, general somatic response and genitourinary response subscale, and in the total SSRS score than the healthy group. Only the depressive disorder group scored significantly higher on the somatic sensitivity subscale than the healthy group, and they also scored significantly higher on the genitourinary response subscale than the anxiety disorder group did. These results suggest that the SSRS is highly reliable and valid, and that it can be effectively utilized as a measure for research of the somatic symptoms related to stress. It also implies that somatic sensitivity and genitourinary responses are associated with depressive disorders. PMID- 16259055 TI - Molecular targets of dietary polyphenols with anti-inflammatory properties. AB - There is persuasive epidemiological and experimental evidence that dietary polyphenols have anti-inflammatory activity. Aspirin and other non-steroidal anti inflammatory drugs (NSAIDs) have long been used to combat inflammation. Recently, cyclooxygenase (COX) inhibitors have been developed and recommended for treatment of rheumatoid arthritis (RA) and osteoarthritis (OA). However, two COX inhibitors have been withdrawn from the market due to unexpected side effects. Because conventional therapeutic and surgical approaches have not been able to fully control the incidence and outcome of many inflammatory diseases, there is an urgent need to find safer compounds and to develop mechanism-based approaches for the management of these diseases. Polyphenols are found in many dietary plant products, including fruits, vegetables, beverages, herbs, and spices. Several of these compounds have been found to inhibit the inflammation process as well as tumorigenesis in experimental animals; they can also exhibit potent biological properties. In addition, epidemiological studies have indicated that populations who consume foods rich in specific polyphenols have lower incidences of inflammatory disease. This paper provides an overview of the research approaches that can be used to unravel the biology and health effects of polyphenols. Polyphenols have diverse biological effects, however, this review will focus on some of the pivotal molecular targets that directly affect the inflammation process. PMID- 16259058 TI - Behcet's disease combined with various types of fistula. AB - Behcet's disease (BD) is a chronic, relapsing, multisystem disorder, characterized by recurrent oral ulcer, genital ulcers, eye lesion, and skin lesion. The underlying pathology is nonspecific vasculitis of all vessel sizes, and severe vasculitis can result in fistula formation of neighboring tissues due to a necrotic process. Herein, eleven cases of BD combined with fistula are presented. In the present study, various types of fistula were associated; enterocutaneous fistula in six patients, and rectovaginal fistula in two. The other three patients showed aortoduodenal fistula, urethrovaginal fistula and urethrocutaneous fistula. They were treated with a corrective operation, but the prognoses were poor due to frequent relapses. PMID- 16259056 TI - Aberrant cell cycle regulation in cervical carcinoma. AB - Carcinoma of the uterine cervix is one of the most common malignancies among women worldwide. Human papillomaviruses (HPV) have been identified as the major etiological factor in cervical carcinogenesis. However, the time lag between HPV infection and the diagnosis of cancer indicates that multiple steps, as well as multiple factors, may be necessary for the development of cervical cancer. The development and progression of cervical carcinoma have been shown to be dependent on various genetic and epigenetic events, especially alterations in the cell cycle checkpoint machinery. In mammalian cells, control of the cell cycle is regulated by the activity of cyclin-dependent kinases (CDKs) and their essential activating coenzymes, the cyclins. Generally, CDKs, cyclins, and CDK inhibitors function within several pathways, including the p16(INK4A)-cyclin D1-CDK4/6-pRb E2F, p21(WAF1)- p27(KIP1)-cyclinE-CDK2, and p14(ARF)-MDM2-p53 pathways. The results from several studies showed aberrant regulation of several cell cycle proteins, such as cyclin D, cyclin E, p16(INK4A), p21(WAF1), and p27(KIP1), as characteristic features of HPV- infected and HPV E6/E7 oncogene-expressing cervical carcinomas and their precursors. These data suggested further that interactions of viral proteins with host cellular proteins, particularly cell cycle proteins, are involved in the activation or repression of cell cycle progression in cervical carcinogenesis. PMID- 16259059 TI - Atopy and house dust mite sensitization as risk factors for asthma in children. AB - Asthma is commonly described as an atopic disease in childhood, but some cases of this disorder do not fit this description. The aim of this study was to evaluate the frequency of atopy, asthma, and sensitization to house dust mites in children with allergic symptoms. This study was performed at the Severance Hospital of Yonsei University with patients who visited the allergy clinic for evaluation of nonspecific upper respiratory symptoms, typical symptoms of asthma, or a general health workup. The patients were divided into three age groups: 0-3 years (group 1), 4-7 years (group 2), and 8-12 years (group 3). Of the 1,244 children examined, 844 (67.8%) were atopic and 400 (32.2%) were non-atopic. The frequency of atopy and asthma increased with age. Asthma was diagnosed in the same proportion (64%) of atopic and non-atopic children. As risk factors for asthma symptoms, the positive values of house dust mite (HDM) sensitivity were significantly increased in groups 1, 2, and 3 to 53.5%, 68.9%, and 80.2%, respectively. A significant difference between the percentage of asthmatics sensitized to HDM and that of asthmatics not sensitized to HDM was found only in group 3. In conclusion, asthma is related to atopy with increasing age, and house dust mite sensitization seems to be an important determinant of asthma in older children in Korea. PMID- 16259060 TI - Real-time CT fluoroscopy (CTF)-guided vertebroplasty in osteoporotic spine fractures. AB - The purpose of this study was to evaluate the clinical feasibility, benefits, and limitations of CT fluoroscopy (CTF)-guided percutaneous vertebroplasty (PVP). PVP under the guidance of CTF without additional guidance by conventional C-arm fluoroscopy was performed in a total of 29 vertebral bodies in 21 patients with vertebral compression fractures. While monitoring sectional CTF images, the needle was advanced from the skin to the target vertebra. Contrast media and polymethylmethacrylate (PMMA) were injected into the target vertebra with careful monitoring of their distribution. After the procedure, an evaluation was conducted to determine whether extraosseous leakage of PMMA occurred and whether sufficient filling of PMMA had been achieved. Needle placement into the target vertebra was easily achieved with both the transpedicular and posterolateral approaches. Injection of PMMA and venous leakage of contrast media were carefully monitored in all patients, and early detection of PMMA leaking was achieved in 5 patients. Extraosseous leakage that had not been detected during the procedure was not found upon postoperative evaluation. Pain scales were significantly decreased after the procedure, and no obvious complications occurred following the procedure. CTF-guided PVP without the combined use of C- arm fluoroscopy was feasible and showed definite benefits. We believe that, in spite of some limitations, CTF-guided PVP provides an alternative technique appropriate in certain situations. PMID- 16259061 TI - The relationship of the anthropometric variables to the infusion rate of rocuronium in the elderly. AB - We have determined the infusion rates of rocuronium in the elderly and young adult patients during sevoflurane and nitrous oxide anesthesia. The correlation of some anthropometric predictors with infusion rate of rocuronium was also investigated for both elderly and young adult. Participating patients were assigned to one of two groups: 1) young adult patients aged 20 to 50 years (n = 30); 2) elderly patients aged over 65 years (n = 30). The anthropometric variables such as height, weight, ratio of weight to body surface area, subscapularis and suprailiac skin folds, body surface area, body mass index and % ideal body weight were evaluated as predictors for infusion rate. The infusion rate in elderly patients was significantly less compared with that in young adult patients (p < 0.05). In elderly patients, no anthropometric predictor was related to the infusion rate of rocuronium. This suggests that the infusion rate of rocuronium for an elderly patient needs to be individualized by monitoring neuromuscular transmission to avoid excessive dose. PMID- 16259062 TI - Percutaneous needle decompression during laparoscopic gastric surgery: a simple alternative to nasogastric decompression. AB - Laparoscopic gastric surgeries are routinely performed with use of a nasogastric tube to decompress the upper gastrointestinal tract. A distended upper gastrointestinal tract can complicate successful laparoscopic gastric surgery as the distention compromises not only the visual field but also the laparoscopic manipulation of the stomach. Since nasogastric intubation is not without risks, we have attempted laparoscopic-assisted gastric cancer surgeries without nasogastric tubes. In this article we describe a simple method of aspirating gastric contents using a 9 cm long 19-gauge needle inserted percutaneously during laparoscopic-assisted gastrectomy. First, a 9 cm long 19-gauge disposable needle was introduced through the abdominal wall. This needle was then introduced to the stomach through the anterior wall and the stomach gases and fluids were aspirated by connecting the needle to suction. Thus, a collapsed upper gastrointestinal tract was easily obtained. We performed this procedure instead of nasogastric decompression on twenty-two patients with gastric cancer who underwent laparoscopic-assisted distal subtotal gastrectomy with lymph node dissection. The results were good with only one patient experiencing wound infection (4.5%) and one patient with postoperative acalculus cholecystitis (4.5%). There were no patients with either intraabdominal infection or anastomotic leakage and none of the patients needed postoperative nasogastric decompression, except the patient who experienced acaculus cholecystitis. Percutaneous needle aspiration is a very simple and efficient technique with little risk of postoperative complications. It can be used as an alternative to nasogastric tube decompression of the gastrointestinal tract for laparoscopic-assisted gastrectomy. PMID- 16259063 TI - Uterine artery Doppler velocimetry during mid-second trimester to predict complications of pregnancy based on unilateral or bilateral abnormalities. AB - We performed this study to evaluate uterine artery Doppler velocimetry (UADV) measurement of unilateral or bilateral abnormalities as a predictor of complications in pregnancy during the mid-second trimester (20-24 weeks). We enrolled 1,090 pregnant women who had undergone UADV twice: once between the 20th and 24th week (1st stage) and again between the 28th and 32nd week (2nd stage) of pregnancy, and then delivered at Yonsei Medical Center. UADV was performed bilaterally. Follow-up UADV was performed between the 28th and 32nd week, and the frequencies of pregnancy-induced hypertension (PIH), fetal growth restriction (FGR), and preterm delivery (before 34 weeks of gestation) were determined. Chi squared and t-tests were used where appropriate, with p < .05 considered significant. According to the results of UADV performed between 20-24 weeks of gestation, 825 women (75.7%) were included in the normal group, 196 (18.0%) in the unilateral abnormality group, and 69 (6.3%) in the bilateral abnormality group. The incidences of FGR were 8.0%, 10.2%, and 26.1%, and the incidences of PIH were 0.1%, 3.6%, and 14.5%, respectively. The incidence of PIH was significantly lower in the normal group. The incidences of preterm delivery were 2.2%, 5.6%, and 8.7%, respectively. PIH developed in 46.7% of patients with bilateral abnormal findings in both the 1st and 2nd stage tests, and developed in none of the patients with normal findings in both tests. Abnormal results found by UADV performed between the 20-24th weeks of pregnancy, such as high S/D ratios regardless of placental location and the presence of an early diastolic notch, were associated with significant increases in the incidences of intrauterine growth restriction (IUGR) and PIH. This was true for both bilateral and unilateral abnormalities. Abnormal findings in bilateral UADV during the second trimester especially warrant close follow up for the detection of subsequent development of pregnancy complications. PMID- 16259064 TI - Comparison of anthropometric data between end-stage renal disease patients undergoing hemodialysis and healthy adults in Korea. AB - Protein-calorie malnutrition is prevalent in hemodialysis (HD) patients. The prevalence of obesity in healthy Korean adults has increased rapidly during the last 10 years. However, there are few large scale data collections available about the current weight status of Korean HD patients. The weight statuses of 10,304 HD patients (data from the Insan Memorial Dialysis Registry 2002, Korean Society of Nephrology) were compared to those of 12,436 control subjects (age > 18) by using body mass index (BMI). Weight status was assessed by WHO classification for Asian-Pacific region [underweight (UW): < 18.5; normal weight (NW): 18.5-22.9; overweight (OW): 23-24.9; obese (OB): 25-29.9; and extremely obese (EOB): > 30 kg/m2] in both the control and HD patients. HD patients had significantly lower body weight and BMI than the controls in all age groups and in both sexes. For the male controls, the proportions of OW and OB showed a reversed U-shape, peaking at the 5th and 6th decades. of the numbers of those classified as NW and UW were relatively small. For the female controls, the proportions of OW and OB progressively increased with age. On the contrary, in HD patients, the proportions of NW and UW were large, up to more than 70%, and those of OW and OB were small in both sexes. In each age group, UW was seen significantly more in the HD group than in the control group. The 6th decade age group showed the highest prevalence ratio for UW in the HD group for both sexes, compared to the controls (Male: 17.33, Female: 17.68). The percentages of UW were related to HD duration and age in both sexes. In conclusion, Korean HD patients seem to have small proportions of OW and OB, compared to the general population, and protein-calorie malnutrition may still be an important nutritional condition. PMID- 16259065 TI - Expression of mucin genes in the human testis and its relationship to spermatogenesis. AB - In this study we investigate the expression pattern of mucin genes in the human testis and evaluate the relationship between the expression of mucin genes and impaired spermatogenesis in the human testis. Thirty human testis tissues were collected from patients undergoing diagnostic testicular biopsy to investigate the cause of infertility. One part of the tissue underwent histological observation, and the other part of the tissue was subjected to semiquantitative RT-PCR of mucin genes, that is, mucin1, 2, 3, 4, and 9. The relative amount of mucin mRNAs was calculated by densitometry using glyceraldehydes-3-phosphate dehydrogenase (GAPDH) as an internal control. The samples were histologically diagnosed as either obstructive azoospermia with normal spermatogenesis (n = 13) or non-obstructive azoospermia with impaired spermatogenesis (n = 17). In the human testis with normal spermatogenesis, mRNA expression of mucin1, 9, 13 and GAPDH were found, but RT-PCR products of mucin 2, 3 and 4 were not detected. In the testis with impaired spermatogenesis, however, RT-PCR product of mucin1 was not found. There was no difference in the other mucin mRNA expression patterns between the testis with either normal or impaired spermatogenesis. To our knowledge, this study is the first that has detected the mRNA of mucin9 and 13 in human testis. This study also shows that mucin1 expression might be closely related to spermatogenesis. Our findings should be substantiated by more direct evidence, such as mucin protein expression and localization. PMID- 16259066 TI - The significance of bladder trabeculation in the female lower urinary system: an objective evaluation by urodynamic studies. AB - This study aimed to investigate the relationship between bladder trabeculation, urinary function, and the stage of pelvic organ prolapse (POP). The medical records of 104 patients with POP who underwent cystoscopies and urodynamic studies were reviewed retrospectively. Age, incidence of detrusor instability, stage and site of POP, and the parameters of urodynamic studies of patients with and without bladder trabeculation were compared. The difference in the incidence of bladder trabeculation was estimated between patients with and without a suspected bladder outlet obstruction. There were significant differences in the patients' age, stage of POP, and maximal voiding velocity. Patients with a suspected bladder outlet obstruction had a significantly higher incidence of bladder trabeculation. In addition, patients with advanced stages of POP were also found to have a higher incidence of bladder trabeculation. PMID- 16259067 TI - The regulators of VEGF expression in mouse ovaries. AB - The objectives of this study were to explore whether ovarian vascular endothelial growth factor (VEGF) expression in mice can be regulated by IL-6 (interleukin-6), angiotensin II, FSH, and hCG; and to test whether the mouse ovarian VEGF expression can result in angiogenesis. The ICR mice were sacrificed, and their ovaries were recovered. Recovered ovaries were treated with IL-6, angiotensin II, FSH, and hCG separately and incubated for 24 hours in alpha-MEM. Expression of mRNA and protein of VEGF were assessed by RT-PCR and immunohistochemistry. The resulting angiogenesis was evaluated through immunohistochemical analysis for CD34. Treatment of mice ovaries with IL-6, FSH, and hCG resulted in a significant increase of VEGF mRNA, and IL-6 was the most potent inducer of VEGF. IL-6 and FSH resulted in increased neovascularization in the follicular phase of mouse ovaries. In contrast, angiotensin II could not increase VEGF expression or neovascularization. We documented an in vitro increase in VEGF expression by IL 6, FSH, and hCG; and reaffirmed that the proliferative response of murine ovarian endothelial cells paralleled an increase of VEGF expression. PMID- 16259068 TI - The involvement of adult stem cells originated from bone marrow in the pathogenesis of pterygia. AB - Pterygium is a proliferative disease. Recent research has reported that stem cells are involved in the pathogenesis of various proliferative diseases, including solid tumors and diabetic proliferate vitreoretinopathy. In previous literature, we hypothesized that adult stem cells originated from bone marrow were involved in the pathogenesis of pterygium. We proved this by immunohistochemical staining with various stem cell markers. The staining showed adult stem cells in the pterygium. c-kit positive cells were observed primarily in the stroma, and some cells were also found in the basal epithelium. AC133 and CD34 positive cells were primarily found in the basal epithelium and were ovoid shaped, similar to the c-kit cells. However, some cells were found in vascular endothelium. STRO-1 positive cells were found mainly in the stroma and were spindle shaped. In recurrent pterygium, cells were more scattered and the expression pattern was denser. Therefore, we suggest a new theory of pterygium pathogenesis. Inflammation caused by environmental factors triggers the abnormal production of some growth factors and cytokines in order to recover from cellular damage. If these healing signals are excessive, limbal basal cells will be changed to abnormally-altered pterygial cells. The excessive wound healing process and remnant altered cells result in recurrence using the same mechanism. PMID- 16259069 TI - Hematopoietic differentiation of embryoid bodies derived from the human embryonic stem cell line SNUhES3 in co-culture with human bone marrow stromal cells. AB - Human embryonic stem (ES) cells can be induced to differentiate into hematopoietic precursor cells via two methods: the formation of embryoid bodies (EBs) and co-culture with mouse bone marrow (BM) stromal cells. In this study, the above two methods have been combined by co-culture of human ES-cell-derived EBs with human BM stromal cells. The efficacy of this method was compared with that using EB formation alone. The undifferentiated human ES cell line SNUhES3 was allowed to form EBs for two days, then EBs were induced to differentiate in the presence of a different serum concentration (EB and EB/high FBS group), or co cultured with human BM stromal cells (EB/BM co-culture group). Flow cytometry and hematopoietic colony-forming assays were used to assess hematopoietic differentiation in the three groups. While no significant increase of CD34+/CD45- or CD34+/CD38- cells was noted in the three groups on days 3 and 5, the percentage of CD34+/CD45- cells and CD34+/ CD38- cells was significantly higher in the EB/BM co-culture group than in the EB and EB/high FBS groups on day 10. The number of colony-forming cells (CFCs) was increased in the EB/BM co-culture group on days 7 and 10, implying a possible role for human BM stromal cells in supporting hematopoietic differentiation from human ES cell-derived EBs. These results demonstrate that co-culture of human ES-cell-derived EBs with human BM stromal cells might lead to more efficient hematopoietic differentiation from human ES cells cultured alone. Further study is warranted to evaluate the underlying mechanism. PMID- 16259070 TI - A case of Chromobacterium infection after car accident in Korea. AB - Chromobacterium violaceum is a gram negative straight rod, 0.8-1.2 by 2.5 to 6.0 m, which is motile by one polar flagella and one to four lateral flagella. The organism inhabits soil and water and is often found in semitropical and tropical climates. Infections in humans are rare. We report a case of infection caused by strains of C. violaceum. A 38-year-old male patient was admitted to KyungHee University Hospital, Seoul, Korea on July 28th, 2003, after a car accident. The patient had multiple trauma and lacerations. He had an open wound in the left tibial area from which C. violaceum was isolated. The strain was resistant to ampicillin, tobramycin, ampicillin/sulbactam, ceftriaxone and cefepime, but was susceptible to amikacin, gentamicin, ciprofloxacin, levofloxacin, trimethoprim/sulfamethoxazole and piperacillin/tazobactam. The patient was treated successfully by debridement, cephapirin sodium and astromicine sulfate. PMID- 16259071 TI - Three cases of diffuse large B-cell lymphoma presenting as primary splenic lymphoma. AB - Primary splenic lymphoma (PSL) is often defined as generalized lymphoma with splenic involvement as the dominant feature. It is a rare disease that comprises approximately 1% of all malignant lymphomas. We investigated three cases of non Hodgkin's splenic lymphoma that had different clinical features on presentation. The patients' survival times from diagnosis ranged from 59 to 143 months, without evidence of relapse after splenectomy and chemotherapy, with or without radiotherapy. This data suggest that PSL is potentially curable. Further studies are needed to evaluate the impact that different treatment modalities without splenectomy have on patient survival. PMID- 16259072 TI - A case of soft tissue myoepithelial tumor arising in masticator space. AB - Soft tissue myoepithelial tumors of the head and neck region are very rare, and only one case of soft tissue myoepithelial tumor occurring in the masticator space has been reported in the world literature. A case of soft tissue myoepithelial tumor with benign histomorphology, but with an invasive growth pattern, occurred in the masticator space of a 46-year- old male patient. Magnetic resonance imaging of paranasal sinus/nasopharynx revealed a well defined, lobulated heterogeneous mass with high signal intensity and dense calcification in the masticator space between the left mandible ramus and pterygoid process. Grossly, the tumor was a well- circumscribed ovoid solid mass and consisted of yellowish gray glistening firm tissue. Histologically, the tumor showed a multinodular growth pattern and consisted of epithelioid cells in chondromyxoid stroma and of spindle-shaped to ovoid cells in the hyaline stroma. The tumor cells appeared bland, and no mitosis or necrosis was found within the tumor. The tumor focally invaded to adhered bone tissue. Immunohistochemically, the tumor cells were diffusely positive for epithelial membrane antigen, smooth muscle actin, but negative for other epithelial markers. Ultrastructurally, the cytoplasm of the tumor cells contained sparse microfilaments and subplasmalemmal densities. Attenuated desmosomes were commonly seen between the tumor cells. PMID- 16259073 TI - Recurrent retroperitoneal cystic lymphangioma. AB - Retroperitoneal cystic lymphangioma is a rare congenital malformation. The majority of lymphangiomas are present at birth and nearly all present before the age of two years. We report a case of giant cystic retroperitoneal lymphangioma in a patient who first presented with symptoms at the age of 7, underwent surgery, and who then suffered a recurrent mass 11 years later. PMID- 16259074 TI - Three cases of Descemet's membrane detachment after cataract surgery. AB - Descemet's membrane detachment (DMD) is an uncommon condition with a wide range of etiologies. More than likely, the most common cause is a localized detachment occurring after cataract surgery. We report three cases of Descemet's membrane detachment that occurred after uncomplicated phacoemulsification cataract surgeries. The first patient was managed without surgical intervention, the second patient was treated using an intracameral air injection, and the last patient was treated with an intracameral perfluoropropane (C3F8) gas injection. All three patients recovered their vision following the reattachment of Descemet's membrane. The three patients were treated according to the extent of the detachment. PMID- 16259075 TI - Barotraumatic perforation of pharyngoesophagus by explosion of a bottle into the mouth. AB - Pharyngoesophageal perforation from an exploding bottle is an extremely rare injury. To date, twenty-four cases have been documented in English literature. In this study, we reported two additional cases of pharyngoesophageal perforation by a bottle exploding in the mouth. Explosion of the bottle occurred when the patients removed the cap of a home-made wine bottle with their teeth, which resulted in pharyngoesophageal perforation. The patients were managed by conservative treatment and operative repair, respectively. Both patients had an uneventful recovery. Possible mechanisms and preventive measures are discussed in this study, along with a review of the literature. PMID- 16259077 TI - Small left atrium: an adjunctive sign of hemodynamically compromised massive pulmonary embolism. AB - Pulmonary embolism (PE) is a common disease with a high mortality rate due to right ventricular dysfunction and underfilling of the left ventricle. We present a case of a 33-year-old man with hemodynamically compromised massive PE. His left atrium was collapsed with marked dilatation of the right atrium and ventricle on multi-detector-row CT scans. The patient was treated with an intracatheter injection of a mutant tissue-type plasminogen activator and subsequently showed clinical and radiological improvements. The small left atrial size in combination with a right ventricular pressure overload was considered to be an adjunctive sign of hemodynamically compromised massive PE. PMID- 16259076 TI - Anomaly of the left anterior descending coronary artery arising from the right sinus of valsalva and ventricular septal defect in adult: a rare case. AB - Anomaly of the left anterior descending (LAD) coronary artery arising from the right sinus of valsalva is frequently seen with tetralogy of Fallot (TOF). The association of the LAD coronary artery with ventricular septal defect (VSD) is uncommon. We described an anomalous origin of the LAD coronary artery from the right sinus of valsalva with ventricular septal defect in a 38-year-old male patient suffering from atypical angina. The LAD coronary artery arose from the right sinus of valsalva, just next to the right coronary artery. There was a single opening in the membranous part of the interventricular septum. From this case, we suggest that angiography is useful for both documenting anomalies of the LAD coronary artery associated with VSD and for determining the safest surgical procedures. PMID- 16259078 TI - Present and future of cancer vaccines. PMID- 16259080 TI - Prognostic significance of the thymidine phosphorylate/dihydropyrimidine dehydrogenase ratio in invasive cervical cancer. PMID- 16259081 TI - Black tea may be as good as green. PMID- 16259082 TI - JavaFit: strange brews. PMID- 16259083 TI - Sunscreen use in children may lower risk of developing future skin cancer. PMID- 16259084 TI - The Delilah revelation. PMID- 16259085 TI - Comment on "real time detection of ventricular fibrillation and tachycardia. PMID- 16259086 TI - Practical ethics. How do you cut benefits? PMID- 16259087 TI - Medication safety issue brief. Medication reconciliation. PMID- 16259089 TI - Health care remains segregated, driving disparities, report asserts. PMID- 16259088 TI - Report raises questions about the future of the hospital safety net. PMID- 16259090 TI - Guide simplifies screening, treating of patients who might drink too much alcohol. PMID- 16259091 TI - Americans concerned about overcrowding and boarding of patients in the ED. PMID- 16259092 TI - Medicare and Medicaid programs; requirements for long term care facilities; nursing services; posting of nurse staffing information. Final rule. AB - In this rule, we finalize provisions specified in the Medicare, Medicaid, and SCHIP Benefits Improvement and Protection Act of 2000 (BIPA) that establish new data collection, posting, and recordkeeping requirements for skilled nursing facilities (SNFs) and nursing facilities (NFs). It requires that on a daily basis for each shift, SNFs and NFs must post nurse staffing data for the licensed and unlicensed staff directly responsible for resident care in the facility. Facility census information must also be posted. This final rule is also part of a broader communication outreach initiative by CMS to provide beneficiaries, their families, and the public with access to updated data and other information that can assist them in making healthcare decisions. PMID- 16259093 TI - [A didactical experience about "paleonutrition"]. AB - The item arises from an "elective" Paleopathology course suggested for students graduating in Medicine and Surgery at the University of Milan. This course discussed subjects as: molecular paleobiology, tumors and infectious paleopathology, paleonutrition, paleopathological museums and history of medicine. The author dealt with the "paleonutrition" subject. Extremely actual aspects emerged which drew the students' attention. PMID- 16259094 TI - [History of psychotherapy]. AB - History of psychotherapy invites to reflect on the evolution of the relation between doctor and patient, that today in official medicine presents some unsatisfactory aspects; in coincidence to this bad relation occurred a success of non conventional medicine, that reserves much space to therapeutical relationship. PMID- 16259095 TI - [The historical bases of a super-specialty: electrocardiography]. AB - In the XVIII century the first structured experiments in the field of bioelectricity were performed, and the Italian scientist Luigi Galvani documented the muscular contraction of a frog undergoing an electric shock. In 1791 he showed that the electric stimulation of the heart of a frog determined the contraction of the heart itself. In the first thirty years of the XIX century galvanometers were developed, and in 1842-42 Carlo Matteucci documented that electric activity was present even in the cardiac muscle at rest. At the end of the XIX century Augustus Waller was among the first scientists to publish an electrocardiographic recording obtained from the human body surface; most of his contemporaneous, however, did not retain that electrocardiography might have been an effective clinical application. Willem Einthoven, instead, was convinced of the widespread feasibility of clinical electrocardiography, and promoted a number of improvements and refinements in electrocardiographic technique. The most important and diagnostic-technical development of electrocardiography occurred in the second half of the XX century, and still today, even if many different sophisticated instrumental examinations are available for cardiologic evaluation, electrocardiography represents an essential first-line diagnostic tool in clinical cardiology. PMID- 16259096 TI - [Eugenio Centanni and the rise of immunology in Italy]. AB - Eugenio Centanni, born in Montotto, in Italy, in 1863, is - together with Mecnikoff and Ehrilich - one of the founders of Immunology. The article presents his main discoveries and underlines the importance of the Italian scientist in contributing to the rise and development of a 'new' discipline. PMID- 16259097 TI - Financial issues in nursing education. PMID- 16259098 TI - Surviving the enrollment growth funding formula. AB - Leadership changes in universities usually bring about philosophical and operational shifts within the institution. When these shifts involve resource allocation procedures, nursing programs are often highlighted because their costs appear great in comparison to traditional academic units. This was the exact situation at East Carolina University, a case history of which is presented here. Strategies for dealing constructively with this negative situation are outlined, as are lessons learned. A positive outcome required a thorough knowledge of funding processes, faculty involvement in generating solution alternatives, frequent and effective communication, and patience. PMID- 16259099 TI - A budget model to determine the financial health of nursing education programs in academic institutions. AB - In the allocation of resources in academic settings, hierarchies of tradition and status often supersede documented need. Nursing programs sometimes have difficulty in getting what they need to maintain quality programs and to grow. The budget is the crucial tool in documenting nursing program needs and its contributions to the entire academic enterprise. Most nursing programs administrators see only an operating expense budget that may grow or shrink by a rubric that may not fit the reality of the situation. A budget is a quantitative expression of how well a unit is managed. Educational administrators should be paying as much attention to analyzing financial outcomes as they do curricular outcomes. This article describes the development of a model for tracking revenue and expense and a simple rubric for analyzing the relationship between the two. It also discusses how to use financial data to improve the fiscal performance of nursing units and to leverage support during times of growth. PMID- 16259100 TI - Contribution margin modeling by academic unit: an evidence-based approach to programmatic decision making. AB - As members of the greater university, it is important that the nursing administrators understand how revenues and expenses are calculated for each academic unit and how higher administration performs comparative analytes. This paper will share the model used by one private comprehensive university to do comparison of contribution margins by each school or college. The model affords an opportunity to ask the "what if" questions surrounding expansion or contractions in programs. PMID- 16259101 TI - A model to identify direct costs of nursing education: the Colorado experience. AB - During spring 2003, national, state, and local economic factors converged in a manner that propelled us to better identify the costs of the educational programs offered within the University of Colorado Health Sciences Center School of Nursing (SON). Two factors prompted analysis of direct costs of the nursing education programs: a shrinking state appropriation and a 36.7% rescission of state funds during that academic year. The SON supports 618 students in four programs (baccalaureate, master of science, doctor of nursing, and doctor of philosophy). During summer 2003, the SON leadership team met numerous times in an iterative process to clarify assumptions and make recommendations in an attempt to cost out academic programs. Data were obtained from a variety of sources. The SON Office of Budget and Finance provided revenue and expense data. The Office of Academic Affairs provided course schedules, the course offering plan, and projected student enrollment in courses. The Division Chairs provided data concerning faculty workload and faculty areas of expertise. The data were compiled in Access tables and arrayed in a series of Excel spreadsheets that captured course data and faculty data. A "what-if" analysis was completed to determine cost of a pilot accelerated baccalaureate program. This method provides a dynamic analytic system shown to be prospectively and retrospectively effective. As a result of this analysis, the following metrics are available: direct cost per student per course/program; revenue per student per course/program; faculty teaching FTE; and faculty-to-student ratio. PMID- 16259102 TI - Managing resources and ensuring accountability: understanding return on investment. AB - Leaders of colleges of nursing face increasing challenges associated with insufficient resources coupled with growing market demands. Managing financial resources and ensuring accountability are essential leadership skills for deans and academic leaders across the nation. An understanding of "return on investment" or "ROI" analysis as a mechanism to negotiate and validate outcomes with varied stakeholders is critical to building a financial business case. ROI analysis is one component of metrics for knowledge management and must be understood from a business perspective. Providing leadership to a resilient college of nursing requires a willingness to utilize innovative strategies and effective metrics to measure outcomes. By raising the effectiveness of base spending, thereby increasing ROI, an academic leader is better positioned to innovate, which is crucial to the future success of nursing education. PMID- 16259103 TI - The dedicated education unit: a practice and education partnership. AB - Both educational institutions and health care agencies have a propensity to look to the past for solutions to their present and projected problems, such as the nursing shortage. One problem that continually surfaces and has become one of two primary limitations on expanding nursing school enrollments is finding and sustaining quality clinical experiences in acute care settings. The demand for these clinical sites continues to grow and the consequent need for effective clinical partnerships grows with it. The Dedicated Education Unit is a creative strategy for addressing the confluence of highly sophisticated marketing efforts promoting the profession of nursing as a whole, the high demand job market for nurses, and the increase in nursing school enrollments at both undergraduate and graduate levels. PMID- 16259104 TI - Benefits and costs of integrating technology into undergraduate nursing programs. AB - Advances in technology over the last decade have resulted in increased opportunities for educators to become more innovative in classroom and clinical teaching. These innovations have allowed students and faculty to access essential clinical information at the point of care/need. By capitalizing on technologies such as personal digital assistants and course delivery shells, faculty and students have both portable and remote access to information that can guide practice and learning activities in clinical, classroom, and distance settings. For instance, a student can use a personal digital assistant to research a patient's new medication at the bedside, study course information, access references during class in response to a question, or download clinical materials from home. Although the benefits of having ready access to information seem obvious, there are costs and strategic planning activities associated with implementing these projects. Clearly, the objective of any academic nursing program is to develop skills among students so they can efficiently access information and use that information to guide their nursing practice. To do so, academic nursing administrators must have the forethought to envision how new technologies can support achieving this goal as well as the ability to put in place the infrastructure supports needed for success. This article presents a case study of how one institution developed the necessary infrastructure and garnished the appropriate resources to implement an ambitious technology initiative integrated throughout a large undergraduate nursing program. In addition, how the integration of technology, online and mobile, can enhance clinical learning will be discussed. PMID- 16259105 TI - The expression of five different claudins in invasive breast carcinomas: comparison of pT1pN1 and pT1pN0 tumors. AB - The evaluation of the role of claudins (CLDNs) in breast carcinogenesis has recently begun. We investigated the expression of CLDNs 1, 2, 3, 4, and 7 in pT1pN0 and pT1pN1 invasive ductal breast carcinomas. Tissue arrays of 30-30 pT1pN0 and pT1pN1 invasive ductal breast carcinomas of different grades were constructed, and the expression of CLDN 1, 2, 3, 4, and 7 proteins was analyzed using standard and immunofluorescent immunohistochemistry. The results were evaluated by light and confocal microscopy. Regarding CLDN 1, 4, and 7 expressions, differences were noted between normal and tumor cells and also between tumors of different grades, while no remarkable differences were noted between pT1pN0 and pT1pN1 tumors. CLDNs 1 and 7 were found to be downregulated in tumor cells compared to the normal epithelium, while CLDN 4 expression was decreased in grade 1 tumors. CLDN 7 protein was abundant in normal epithelia, and the staining decreased in grade 3 tumors. There were no differences between normal and neoplastic cells regarding CLDN 2 and 3 expressions. As a preliminary result, our observations suggest that the analyzed CLDNs do not promote tumor metastasis. On the basis of our findings, it seems that CLDN 1, CLDN 4, and CLDN 7 may rather have an important role in tumorigenesis or in cell-to-cell adhesion. PMID- 16259106 TI - Caveolin-1 and 14-3-3 sigma expression in follicular variant of thyroid papillary carcinoma. AB - There are two prominent types of thyroid carcinoma, papillary carcinoma (PC) and follicular carcinoma (FC) arising from thyroid follicular cells, which have different biological characteristics. The follicular variant of papillary carcinoma (FVPC) has a structure similar to FC, although it is classified as a subtype of PC. We have previously demonstrated that caveolin-1, a major component of caveolae and an inhibitor of growth signal transduction, and 14-3-3 sigma, a regulator of the cell cycle and signal transduction, are frequently expressed in pure PC, but not in FC. In this study, we investigated these expressions in FVPC and FC. Caveolin-1 and 14-3-3 sigma were expressed in 96.2% and 92.3% of FVPC, respectively. However, almost all FC, with only a few exceptions, were negative for these findings, indicating that these proteins were more frequently expressed in FVPC than in FC (p < 0.0001). These findings suggest that caveolin-1 and 14-3 3 sigma positively or negatively regulate the development of FVPC but not that of FC, reflecting the difference of biological characteristics of these two types of carcinoma. PMID- 16259108 TI - Loss of heterozygosity of APC and CDH1 genes in laryngeal squamous cell carcinoma. AB - The molecular mechanisms involved in the development and progression of laryngeal cancer, specifically squamous cell carcinoma, still need further investigation and elucidation. Twenty-two laryngeal squamous cell carcinomas were analyzed in our study regarding genetic changes of two tumor suppressor genes: Adenomatous polyposis coli (APC) and E-cadherin (CDH1). APC gene instability was tested by polymerase chain reaction (PCR)/loss of heterozygosity (LOH) using the restriction fragment length polymorphism (RFLP) method. The samples were also screened for mutations using the heteroduplex method. E-cadherin gene was analyzed by PCR amplification of tetranucleotide marker (D16S752) linked to E cadherin gene. The results of our analysis showed three samples with LOH of the APC gene out of 15 heterozygous patients (20%). Only one LOH of the CDH1 gene (5.5%) out of 18 heterozygous patients was discovered. D16S752 marker did not reveal any replication error-positive samples. There were six samples showing heteroduplexes (33%) encompassed in APC's exon 11. Altogether, nine samples (41%) showed alterations of the APC gene. Our results suggest that alterations of APC gene may have a role in squamous cell carcinoma development. Detected LOH of the E-cadherin gene indicates that genetic changes of this gene are not very frequent, but that other components of the wnt signaling cascade may also be involved. PMID- 16259107 TI - S100A9 expression is significantly linked to dedifferentiation of thyroid carcinoma. AB - S100A9, a calcium-binding protein, is associated with myeloid cell differentiation and is expressed in some adenocarcinomas as well as in squamous epithelia and squamous cell carcinoma. In this study, we immunohistochemically investigated S100A9 expression in thyroid neoplasms. S100A9 was absent in normal follicles, follicular adenoma, and follicular and papillary carcinomas with conventional growth structures. In lesions showing a solid, trabecular, or scirrhous growth pattern, S100A9 immunoreacitivity was occasionally observed. One (5.9%) of the 17 follicular carcinomas and three (7.8%) of the 38 papillary carcinomas were regarded as positive for S100A9, but the positive cell areas always accounted for 5% or less. However, S100A9 was positive in all 19 undifferentiated carcinomas examined. Among them, the positive cell area was greater than 5% in 16 (84.2%), and greater than 25% in six (31.6%) cases. It is therefore suggested that S100A4 protein plays an important role in thyroid carcinoma dedifferentiation, and can be considered a novel characteristic of undifferentiated carcinoma. PMID- 16259109 TI - Cholangitis: a histologic classification based on patterns of injury in liver biopsies. AB - Inflammatory disorders of the biliary tract present difficult diagnostic problems in liver needle biopsies. The aim of this study was to perform a detailed histologic analysis of liver biopsies from patients with biliary tract disorders, classify them by pattern of inflammation, and determine the accuracy of the histologic classification by clinical follow-up. Percutaneous liver needle biopsies from the surgical pathology files of UmassMemorial Healthcare (UMMHC) from 2000 to 2003 with a diagnosis suggesting a biliary tract process (n = 32) and four biopsies from cases with systemic non-biliary tract disorders were analyzed for multiple histologic features and classified as one of five patterns: acute cholangitis/pericholangitis (ACP), lymphocytic cholangitis (LC), granulomatous (G), ductopenia (D), or non-specific (NS). When compared to the "gold standard" diagnosis based on all clinical data, the concordance between the histologic classification and the clinical diagnosis was: 50% for ACP and bile duct obstruction; 77% for LC and immune-mediated cholangitis NOS; 100% for G and G cholangitis; 100% for D and idiopathic adulthood D; and 50% for NS and non biliary tract disorders. Our findings suggest that classifying biopsies by pattern of injury is helpful in guiding the subsequent clinical work-up. ACP pattern correlates with bile duct obstruction, infection, and ischemia. LC correlates with serologic studies supporting immune-mediated processes. G pattern suggests further work-up for PBC, drug, tuberculosis, or sarcoidosis. D pattern establishes the clinical diagnosis. NS pattern includes cases of primary sclerosing cholangitis, which cannot be diagnosed by biopsy alone. PMID- 16259110 TI - Expression of CDX2 and MUC2 in Barrett's mucosa. AB - Barrett's mucosa is a risk factor for esophageal adenocarcinoma and should be detected at an early stage. It is defined by the presence of columnar epithelium with goblet cells in the lower esophagus, but histologic diagnosis can be uncertain in the absence of distinct goblet cells. We investigated 55 biopsies from 48 patients with endoscopically plain Barrett's esophagus and performed immunohistochemistry for CDX2 and MUC2. In addition, alcian blue (pH 2,5)/PAS staining was done. In histologically unequivocal Barrett's mucosa, nuclear expression of CDX2 in goblet cells and many columnar cells, as well as cytoplasmic positivity for MUC2 in goblet cells, could be observed. Alcian blue (pH 2,5)/PAS stained acidic mucins in goblet cells and in some non-goblet columnar cells. In six cases, no definite Barrett's mucosa was present, and no expression of MUC2 could be observed. In these biopsies, there was granular cytoplasmic and/or focal nuclear staining for CDX2 in non-goblet columnar epithelial cells, indicating their intestinal differentiation. We suggest that this peculiar mucosa is the precursor of unequivocal Barrett's mucosa and would designate it early Barrett's mucosa. Alcian blue for acidic mucins is inconsistent in this epithelium and does not reliably indicate early intestinal differentiation. PMID- 16259111 TI - Napsin A is useful to distinguish primary lung adenocarcinoma from adenocarcinomas of other organs. AB - The aim of the present study was to compare the usefulness of the peripheral airway cell markers, naspin A and surfactant protein A (SP-A), for distinguishing primary lung adenocarcinoma from adenocarcinomas of other organs in various clinical conditions. Immunohistochemical expression of napsin A and SP-A was analyzed at primary sites of 120 lung carcinomas and 40 adenocarcinomas of other organs, at lung metastatic sites of 32 adenocarcinomas of other organs, and in metastatic lymph nodes of 21 lung adenocarcinomas and 45 adenocarcinomas of other organs. Napsin A and SP-A expressions were compared between primary and recurrent sites in 8 lung adenocarcinomas. Napsin A and SP-A expressions were noted in 84.3% and 53.0% of primary sites of 83 lung adenocarcinomas, respectively, but neither napsin A nor SP-A was expressed at primary sites of other histological types of lung carcinomas or at primary or metastatic sites of adenocarcinomas of other organs. In lung adenocarcinomas, napsin A and SP-A were expressed in metastatic lymph nodes in 81.0% and 19.0%, respectively, and at recurrent sites in 87.5% and 37.5%, respectively. Napsin A is superior to SP-A for distinguishing primary lung adenocarcinoma from adenocarcinomas of other organs at primary, metastatic, and recurrent sites. PMID- 16259112 TI - Evaluation of nucleolar organizer region (NOR) parameters in the uterine leiomyoma. AB - Nucleolar organizer regions (NORs) were assessed in 27 women affected by uterine leiomyoma. Tissue samples obtained during surgery were silver-stained according to the method of Ploton et al. The assessed parameters were as follows: the number of argyrophylic nucleolar organizer regions (AgNORs) per nucleus, single AgNOR area, AgNOR intranuclear distribution, and AgNOR coefficient. The parameters were assessed quantitatively. It was found that the AgNOR coefficient was higher in uterine leiomyoma compared to the normal smooth muscle cells. The development of leiomyoma is associated with a marked decrease in myocyte nucleolar area, which accounts for 30% of the nucleus in the normal myometrium. With the comparable AgNOR number in the single nucleus, there were no differences in the single AgNOR granule area (1.21 microm2 +/- 0.047 and 1.11 microm2 +/- 0.025 in normal myometrium and in leiomyoma, respectively). In the normal myometrium, there was a positive correlation between nuclear area and the single AgNOR granule area, as well as between the AgNOR coefficient and the single AgNOR granule area. There was also a negative correlation between the number of granules per nucleus and their central and peripheral intranuclear distribution. The development of leiomyoma was associated with loss of all correlations observed in the control group. PMID- 16259113 TI - Vascular endothelial growth factor expression in untreated and androgen-deprived patients with prostate cancer. AB - The aim of the study was to investigate immunohistochemically the expression of vascular endothelial growth factor (VEGF) in untreated and androgen-deprived patients with prostate cancer. The study included 20 patients with prostate cancer who had undergone transurethral prostatectomy due to infravesical obstruction. All patients had been receiving androgen deprivation therapy for at least 3 months. Transurethral prostatectomy specimens were examined for VEGF expression after androgen deprivation, and the biopsy samples of the same patients were used for the evaluation of VEGF expression before androgen deprivation. VEGF expression was analyzed using immunohistochemistry. Staining patterns determined by the staining scores were compared before and after treatment. The correlation of VEGF expression with PSA, Gleason score, and the percent change in PSA after treatment was also investigated. Eligible biopsy specimens were available in 15 of the 20 patients, allowing for the evaluation of VEGF expression before treatment. All prostate cancer specimens were positive. VEGF was localized mainly in the cytoplasm or on the membrane of carcinoma cells. Staining was strong in 86.7% of patients before androgen deprivation. Heterogeneous staining (strong in 25%, moderate in 35%, and weak in 40%) was observed after treatment. Staining scores were significantly higher in patients before androgen deprivation and showed a significant decrease after androgen deprivation (p = 0.007). Tumor staining correlated with Gleason score. No significant correlation was determined between VEGF expression and pre-treatment PSA and percent change of PSA after treatment. Immunohistochemical results indicate that VEGF expression is downregulated by androgen deprivation therapy. VEGF may be a potential target for therapeutic intervention in prostate cancer. PMID- 16259114 TI - Tissue microarrays from HOPE-fixed specimens allow for enhanced high throughput molecular analyses in paraffin-embedded material. AB - In this study, we describe a technique that allows for the production and the use of tissue microarrays (TMA) from HOPE-fixed, paraffin-embedded specimens. The combination of these two technologies unites the advantages of the high throughput aspects in TMA, with good preservation of nucleic acids, proteins, and morphology of HOPE-fixed tissues, thus substantially widening the capabilities presently available for molecular studies in paraffin-embedded tissues. PMID- 16259115 TI - A case report of dystrophic localized amyloidosis that developed in the left thigh. AB - We report on a 50-year-old man with dystrophic localized amyloidosis who noticed a soft tumor in his left thigh about 20 years ago, after which the tumor has gradually enlarged. The multicystic tumor showed hemorrhage, hematoma, necrosis, fibrosis, and tiny nodules and various polymorphous granulomas were observed. One was rich in eosinophilic amorphous materials and cholesterol crystals, and was poor in cell reaction. Another was formed by granuloma consisting of multinucleated giant cells, foamy cells, and macrophages. Transitional granulomas between the two were also observed. The materials showed eosinophilia and red staining and apple-green birefringence in polarized light by alkaline Congo-red stain, and they were also resistant to potassium permanganate pretreatment. They were also positive for amyloid P component and consistently negative for amyloid A, kappa- and lambda-light chains, beta2-microglobulin, and transthyretin. Therefore, it was suggested that this might be an amyloid derived from the hematoma, which has not been reported to date. PMID- 16259116 TI - Malignant stromal tumor, so called "gastrointestinal stromal tumor", with rhabdomyomatous differentiation occurring in the gallbladder. AB - Gastrointestinal stromal tumors (GISTs) constitute the largest category of primary nonepithelial neoplasms of the gastrointestinal tract. It is extremely rare that this tumor occurs in the bile tract, and only a few cases have been reported. Immunohistochemically, the tumor cells revealed a phenotype similar to Cajal cells, occasionally with differentiation to smooth muscle cells or neural cells. We present a case of malignant stromal tumor similar to GISTs with rhabdomyomatous differentiation of the gallbladder in a 68-year-old woman. The resected tumor was predominantly composed of spindle cells with rhabdomyomatous differentiation. Immunohistochemical study revealed diffuse staining of tumor cells using vimentin despite negative staining for desmin or S-100. This indicated a mesenchymal origin of the cells without smooth muscle or neuronal differentiation. Myoglobin-positive cells, in which phosphotungstic acid hematoxylin staining revealed cross striations of the cytoplasm, suggested rhabdomyomatous differentiation. Diffuse positivity for KIT in the cells suggested that the pathogenesis of this tumor may resemble that of GIST. The tumor may have derived from a mesenchymal stem cell that had undergone partial rhabdomyomatous differentiation. PMID- 16259117 TI - Pseudosaccharin amine derivatives: synthesis and elastase inhibitory activity. AB - Pseudosaccharin amines were synthesized from saccharin either by the reaction of pseudosaccharin chloride with amines, or via thiosaccharin which was treated with amines yielding thiosaccharinates, and their reaction with glacial acetic acid. This route gave lower yields than the first way. The synthesis of alkyl [(1,1 dioxo-benzo[d]isothiazol-3-yl)amino]alkanoates as possible Human Leukocyte Elastase (HLE) inhibitors was realized by the reaction between amino acid esters and pseudosaccharin chloride. Hydrolysis of the esters was possible under aqueous basic conditions. Selected compounds were screened for elastase inhibitory activity. Compounds 4k and 4m were found to be reversible inhibitors of HLE with Ki values of 45 microM and 60 microM. PMID- 16259118 TI - [Synthesis of substituted 6-phenylpyrazolo[3,4-d]pyrimidines with potential adenosin-A2A antagonistic activity]. AB - Substituted pyrazolo[3,4-d]pyrimidines were prepared by reaction of methyl- or phenylhydrazine with pyrimidine derivatives containing a methylthio or chlorine substituent as nucleofuge. Using a methylthio-6-imino-1,3-thiazine as starting material instead of a methylthiopyrimidine the conversion with phenylhydrazine could already be achieved under mild conditions thus leading first to the formation of a hydrazinopyrimidine. The affinity of the products against the human adenosine A2A receptor was determined. PMID- 16259119 TI - Synthesis of zinc-crosslinked thiolated alginic acid beads and their in vitro evaluation as potential enteric delivery system with folic acid as model drug. AB - The aim of this study is to explore the potential of synthetic modifications of alginic acid as a method to enhance the stability of its complexes with divalent cations under physiological conditions. A fraction of algin's carboxylic acid moieties was substituted with thiol groups to different substitution degrees through conjugating alginate to cysteine to produce alginate-cysteine (AC) conjugates. Infrared spectrophotometry and iodometry were used to characterize the resulting polymeric conjugates in terms of structure and degree of substitution. Moreover, zinc ions were used to crosslink the resulting AC polymers. Folic acid loaded beads were prepared from Zinc-crosslinked AC polymers (AC-Zn) of different cysteine substitution degrees. The generated beads were then investigated in vitro for their capacity to modify folic acid release. AC-Zn polymeric beads resisted drug release under acidic conditions (pH 1.0). However, upon transfer to a phosphate buffer solution (pH 7.0) they released most of their contents almost immediately. This change in drug release behavior is most probably due to the sequestering of zinc cations by phosphate ions within the buffer solution to form insoluble chelates and, to a lesser extent, the ionization of the carboxylic acid and thiol moieties. Removal of zinc ions from the polymeric matrix seems to promote polymeric disintegration and subsequent drug release. A similar behavior is expected in vivo due to the presence of natural zinc sequestering agents in the intestinal fluids. AC-Zn polymers provided a novel approach for enteric drug delivery as drug release from these matrices complied with the USP specifications for enteric dosage forms. PMID- 16259120 TI - Quality control of tablets by Near Infrared (NIR)-Spectroscopy. AB - Today, NIR-spectroscopy is an established analytical technique not only in the identification of raw materials but also in the quantification of active ingredients in tablets. In this work calibration models were set up with tablets of the same active ingredient but of miscellaneous origin and manufacturess. Consequently the tablets had different excipients and appearance. The pharmaceutical preparations used included atenolol 100 mg tablets, enalapril 20 mg tablets and acetylsalicylic acid (ASS) tablets of different dosage units. In order to proof if the calibration models set up are generally feasible the assay declared by the manufacturer was used to calculate the partial least square (PLS) calibration. With respect to enalapril tablets simultaneous analysis by HPLC, according to USP 26 was carried out. It was investigated if such methods allow a determination of active ingredients in tablets within limits of +/- 10% of declaration. It was shown that it is possible to set up calibration models to quantify active ingredients in tablets independent of adjuvants or optical appearance. Additionally it could be shown that NIR-spectroscopy is also applicable to determine the concentration of active ingredients in blister-packed tablets. PMID- 16259121 TI - Electrochemical study of natamycin--analytical application to pharmaceutical dosage forms by differential pulse voltammetry. AB - The electrochemical oxidation and determination of natamycin has been carried out at a carbon paste electrode in aqueous solutions in the pH range of 2.5-10.30 by cyclic and differential pulse voltammetry. Best results were obtained with the differential pulse voltammetric technique in 0.5 M sulfuric acid at pH 1.82. The diffusion controlled nature of the waves was established. A differential pulse voltammetric technique for the determination of natamycin 0.5 M sulfuric acid which allows quantitation over the range of 2 x 10(-6)-8 x 10(-5) M range method is proposed. Limit of detection and limit of quantitatification were 1.5 x 10(-6) and 5 x 10(-6) M, respectively. Based on this study a simple, rapid, selective and sensitive voltammetric method was developed for the determination of natamycin in capsules. In order to validate the proposed method, UV spectroscopy was applied. PMID- 16259122 TI - Skin moisturizing effect and skin penetration of ascorbyl palmitate entrapped in solid lipid nanoparticles (SLN) and nanostructured lipid carriers (NLC) incorporated into hydrogel. AB - This study was performed as a complimentary to our previous study regarding the chemical stability of ascorbyl palmitate (AP) in solid lipid nanoparticles (SLN), nanostructured lipid carriers (NLC) and for comparison, in nanoemulsion (NE) incorporated into a hydrogel produced by high pressure homogenization. AP is known as an effective antioxidant that protects tissue integrity similar to vitamin C. Recently, its moisturizing activity in conventional topical formulations was found to be high. The aim of the present study was to investigate the moisturizing potential of AP in SLN and NLC incorporated into hydrogel as colloidal carrier systems. It has been known that SLN and NLC have occlusive effects, but AP incorporation moisturized skin significantly better than placebo in short-term (p < 0.001) and long-term trials (p < 0.01) for both SLN and NLC. In the second part of the study, SLN and NLC were found to sustain the penetration of AP through excised human skin about 1/2 and 2/3 times compared to NE (p < 0.001 and p < 0.01), respectively, due to the solid state of Witepsol E85 in the lipid phase. PMID- 16259123 TI - In vitro methods for the evaluation of drug availability from suppositories: comparison between biological and artificial membranes. AB - Drug availability from suppositories is currently evaluated in vitro by means of a model consisting of a dialysis tube (porous membrane) or isolated biological membrane (animal rectum). We propose a new alternative in vitro method to determine drug availability from suppositories consisting of an artificial membrane soaked with n-octanol, coupled with a filter paper sheet soaked with phosphate buffer. This method provides for an integrated hydro-lipophilic simulation of the biological membrane, including the mucus layer adhering to the rectal mucosa. By simply using the porous membrane, the amount of drug released varied directly according to its solubility for formulations with lipophilic excipients. For formulations with hydrophilic excipients, drugs with low/intermediate solubility in water showed increased availability in comparison to lipophilic excipients. The in vitro rat rectum model provided overall results that were similar to those obtained with the porous membrane method, although the percentage values of AUC were lower. The new model of in vitro simulated absorption produced a degree of drug availability that was lower in comparison to both previous methods. However, the simulated model appeared to give a pattern of drug availability closer to that of the model of in vitro rat rectum. The new in vitro artificial model thus appears to be useful in suppositories preformulation studies, allowing for an estimate of drug availability and the choice of the most adequate excipient. PMID- 16259124 TI - Interaction between ethanol and diazepam in mice: chronobiological aspects. AB - The mechanism of action of benzodiazepines and ethanol demonstrates that these agents can synergistically affect the central nervous system (CNS). The effects of both ethanol and diazepam are likely to depend on the time of the day when they were administered. Diazepam influence on ethanol-induced sleeping and hypothermic activity in mice as well as the influence of combined administration of these agents on spontaneous locomotor activity and coordination in mice (rota rod) were investigated. Experiments were carried out in the light phase (10:00 12:00 h) and the dark phase (22:00-24:00 h). It was shown that ethanol-induced sleeping time was longer in the dark phase than the light phase, and that ethanol increased spontaneous locomotor activity both in the light and the dark. Ethanol induced hypothermia was lower in the dark than in the light. Diazepam decreased locomotor activity more strongly in the dark phase than by day. It impaired the hypothermic action of ethanol in the light phase, but did not have such an effect in the dark phase. Diazepam prolonged ethanol-induced sleep in the light phase, enhanced its action on locomotor coordination and decreased the stimulating effect of ethanol on spontaneous locomotor activity in mice. The chronobiological effect of the interaction between diazepam and ethanol seems to be of practical importance (sleep and motor coordination). PMID- 16259125 TI - Vitamin K3 triggers human leukemia cell death through hydrogen peroxide generation and histone hyperacetylation. AB - Vitamin K3 (VK3) is a well-known anticancer agent, but its mechanism remains elusive. In the present study, VK3 was found to simultaneously induce cell death, reactive oxygen species (ROS) generation, including superoxide anion (O2*-) and hydrogen peroxide (H2O2) generation, and histone hyperacetylation in human leukemia HL-60 cells in a concentration- and time-dependent manner. Catalase (CAT), an antioxidant enzyme that specifically scavenges H2O2, could significantly diminish both histone acetylation increase and cell death caused by VK3, whereas superoxide dismutase (SOD), an enzyme that specifically eliminates O2*-, showed no effect on both of these, leading to the conclusion that H2O2 generation, but not O2*- generation, contributes to VK3-induced histone hyperacetylation and cell death. This conclusion was confirmed by the finding that enhancement of VK3-induced H2O2 generation by vitamin C (VC) could significantly promote both the histone hyperacetylation and cell death. Further studies suggested that histone hyperacetylation played an important role in VK3 induced cell death, since sodium butyrate, a histone deacetylase (HDAC) inhibitor, showed no effect on ROS generation, but obviously potentiated VK3 induced histone hyperacetylation and cell death. Collectively, these results demonstrate a novel mechanism for the anticancer activity of VK3, i.e., VK3 induced tumor cell death through H2O2 generation, which then further induced histone hyperacetylation. PMID- 16259126 TI - Lipoic acid improves glucose utilisation and prevents protein glycation and AGE formation. AB - The present study investigates the antiglycating effect of alpha-lipoic acid (LA) in high fructose-fed rats in vivo and its potential to inhibit the process of glycation in vitro. In addition, the effect of LA on glucose utilisation in rat diaphragm was also studied. Rats fed a high fructose diet (60% total calories) were administered with 35 mg/kg b.w, lipoic acid (LA) intraperitoneally for 20 days. The effects of LA on plasma glucose, fructosamine, protein glycation and glycated haemoglobin in high fructose rats and on in vitro glycation were studied. In vitro utilization of glucose was carried out in normal rat diaphragm in the presence and absence of insulin in which LA was used as an additive. The contents of glucose, glycated protein, glycated haemoglobin and fructosamine were significantly lowered on LA administration to high fructose-fed rats. LA prevented in vitro glycation and the accumulation of advanced glycation end products. Further LA enhanced glucose utilization in the rat diaphragm. This effect was additive to that of insulin and did not interfere with the action of insulin. The findings provide evidence for the therapeutic utility of lipoic acid in diabetes and its complications. PMID- 16259127 TI - Antimicrobial, cytotoxic and antioxidant activity of selected basidiomycetes from Yemen. AB - Dichloromethane, methanol and aqueous extracts of 23 selected Basidiomycetes species fruiting bodies collected in Yemen were screened in vitro for their antibacterial activities against three Gram-positive bacteria (Staphyloccocus aureus, Bacillus subtilis, Micrococcus flavus), two Gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa) and against one yeast fungus (Candida maltosa), as well as for their cytotoxic and antioxidant activity. The highest antibacterial activity was shown by extracts from Agaricus sp. (Type 1), Coriolopsis caperata, Ganoderma colossus, Ganoderma resinaceum, Phellorinia herculea and Tulostoma obesum. Strong antioxidative effects employing the DPPH assay were exhibited by methanol extracts from Ganoderma resinaceum, Inonotus ochroporus, Phellinus rimosus and Phellorinia herculea. The results provide evidence that some of the studied fungi might be potential sources for new biologically active agents. PMID- 16259128 TI - Screening of Leishmania APRT enzyme inhibitors. AB - Adenine phosphoribosyltransferase (APRT) enzyme from Leishmania tarentolae has been proposed as a target for the rational search of new leishmanicidal drugs. In this paper, we describe the evaluation of the inhibitory activity on L. tarentolae APRT enzyme of 46 crude extracts of Meliaceae and Rutaceae plants, besides three furoquinolone alkaloids. The results showed that 21 extracts were able to decrease the APRT enzymatic activity (IA% > or = 50). The methanolic extracts from roots and leaves of Cedrela fissilis and from fruits, branches and leaves of Cipadessa fruticosa have showed strong activities. Therefore, these species could be a promising source of lead compounds for the rational design of new leishmanicidal drugs. The phytochemical investigation of an active fraction from Almeidea rubra afforded the alkaloids isodutaduprine, isoskimmianine and isokokusagine, which showed low to moderate activity on APRT. PMID- 16259129 TI - New aristolochic acid, aristololactam and renal cytotoxic constituents from the stem and leaves of Aristolochia contorta. AB - Two novel phenanthrene derivatives, aristololactam IVa (1) and 9-hydroxy aristolochic acid I (2) were isolated from the stem and leaves of Anstolochia contorta Bunge, together with 17 known compounds (3-19). The structures of these compounds were determined by spectroscopic analysis. The phenanthrenes obtained were tested for cytotoxicity against renal proximal tubular epithelial cell line (HK-2). Aristololactam IVa and 7-methoxy aristololactam IV were found to have strong cytotoxic activity against HK-2 cells with a potency similar to or even stronger than those of aristolochic acid I and aristololactam I. PMID- 16259130 TI - A new biflavonoid from Aristolochia contorta. AB - From the fruits of Aristolochia contorta Bge, beside the known aristolochic acids IVa and VII, aristolactam-N-beta-D-glucopyanoside, aristoloctam Ia N-beta-D glucopyanoside, pinitol and daucosterol, a new biflavonoid was isolated. Its structure was determined as (+/-)-2"R,3"R-dihydro-3"-hydroxyamentoflavone (1) by means of spectral methods including 1D-, 2D-NMR and HR-ESIMS and the known compounds were identified on the basis of comparing their NMR data with those of the corresponding compounds in the literature. PMID- 16259131 TI - Spectrofluorimetric determination of fendiline in tablets. AB - A simple, sensitive and accurate spectrofluorimetric method was developed for the assay of fendiline in tablets. Validation of the method provided good results concerning linearity, precision and accuracy. The linearity range was found to be 0.1-0.8 mg/l. The fluorescence intensity was measured at 288 nm for fendiline tablet solutions. It was also found that the excipients in the commercial tablets did not interfere with the method. PMID- 16259132 TI - Topical delivery of retinyl ascorbate co-drug: 6. Determination of toxic dose and antioxidant activity in cultured human epidermal keratinocytes. AB - The maximum effective dose of retinyl ascorbate and its potential therapeutic benefits against induced oxidative damage were assessed in vitro using cultured human epidermal keratinocytes. RA-AsA exhibited toxic effects at concentrations >6 microM. The findings indicate to the potency of RA-AsA as free radical scavenger and cell proliferation regulator. PMID- 16259133 TI - Effect of different phenolic compounds on alpha-amylase activity: screening by microplate-reader based kinetic assay. AB - The inhibitory effect of different polyphenolic plant compounds on alpha-amylase activity was investigated in vitro. A kinetic assay was performed using 96-well plates. Acarbose was used as positive control (IC50: 23.2 microM). Some of the tested compounds, occurring in plants traditionally used in anti-diabetic tea species, showed an inhibition of the enzyme in physiological concentrations, e.g. luteolin, tannic acid, and isochlorogenic acid. PMID- 16259134 TI - A new phenylethanoid glycoside from Clerodendrum inerme. AB - A new phenylethanoid glycoside, 2-(3-methoxy-4-hydroxylphenyl) ethyl-O-2",3" diacetyl-alpha-L-rhamnopyranosyl-(1-->3)-4-O-(E)-feruloyl-beta-D-glucopyranoside, was isolated from the aerial parts of Clerodendrum inerme (L.) Gaertn, together with monomelittoside, melittoside, inerminoside A1, verbascoside, isoverbascoside, campneoside I. Their structures were determined by spectroscopic methods. PMID- 16259135 TI - Pay for MAHOASHC. PMID- 16259136 TI - Better business, more money. PMID- 16259137 TI - Deals on wheels. PMID- 16259138 TI - The new frontier of data mining. PMID- 16259139 TI - Five trends impacting hospitals' business processes and financial health. PMID- 16259140 TI - An ideal dics storage solution. PMID- 16259141 TI - What works. Managing the business of surgery. New York hospital employs perioperative system to increase revenue. PMID- 16259143 TI - Physician practice management hotlist. PMID- 16259142 TI - What works. Meeting security regulations for e-mail. Texas health system selects e-mail encryption solution. PMID- 16259144 TI - Flying in the dark: physicians and pay for performance. PMID- 16259145 TI - Research in mental health: social etiology versus social consequences. AB - This article differentiates a social etiology model focused on identifying the social antecedents of one particular mental disorder from a social consequences model concerned with the overall mental health consequences of various social arrangements. In the social etiology model, people with disorders other than the one particular disorder singled out for investigation are implicitly classified as "well." This disorder-specific model is inappropriate for the more general sociological task of identifying the consequences of various social arrangements, such as concentrated poverty, racial segregation, and gender stratification. It is problematic because these consequences are typically nonspecific, not limited to one particular disorder. From this perspective, persons classified as "well" in the disorder-specific model who have a different disorder are misclassified. Consequently, the impact of social arrangements is underestimated, and estimates of causal effects are biased. To address these problems, the full range of theoretically derived mental health outcomes needs to be simultaneously analyzed. PMID- 16259147 TI - Is obesity stigmatizing? Body weight, perceived discrimination, and psychological well-being in the United States. AB - We investigate the frequency and psychological correlates of institutional and interpersonal discrimination reported by underweight, normal weight, overweight, obese I, and obese II/III Americans. Analyses use data from the Midlife Development in the United States study, a national survey of more than 3,000 adults ages 25 to 74 in 1995. Compared to normal weight persons, obese II/III persons (body mass index of 35 or higher) are more likely to report institutional and day-to-day interpersonal discrimination. Among obese II/III persons, professional workers are more likely than nonprofessionals to report employment discrimination and interpersonal mistreatment. Obese II/III persons report lower levels of self-acceptance than normal weight persons, yet this relationship is fully mediated by the perception that one has been discriminated against due to body weight or physical appearance. Our findings offer further support for the pervasive stigma of obesity and the negative implications of stigmatized identities for life chances. PMID- 16259146 TI - Race differentials in obesity: the impact of place. AB - This article reveals race differentials in obesity as both an individual- and neighborhood-level phenomena. Using neighborhood-level data from the 1990-1994 National Health Interview Survey, we find that neighborhoods characterized by high proportions of black residents have a greater prevalence of obesity than areas in which the majority of the residents are white. Using individual-level data, we also find that residents of neighborhoods in which at least one-quarter of the residents are black face a 13 percent increase in the odds of being obese compared to residents of other communities. The association between neighborhood racial composition and obesity is completely attenuated after including statistical controls for the poverty rate and obesity prevalence of respondents' neighborhoods. These findings support the underlying assumptions of both institutional and social models of neighborhood effects. PMID- 16259148 TI - A changed America? The effects of September 11th on depressive symptoms and alcohol consumption. AB - In the weeks following the terrorist attacks of September 11, 2001, social commentators argued that America had profoundly "changed." In light of these arguments and the literature on disasters, we examine the immediate and longer term mental health consequences of September 11th using a national sample of full time American workers. We model the effects of temporal proximity to the attacks on depressive symptoms and alcohol consumption, while controlling for demographic characteristics. Our data revealed a significant increase in the number of depressive symptoms reported during the 4 weeks after the attacks. In the subsequent weeks, levels of depressive symptoms returned to pre-September 11th levels. Contrary to expectations, there was some indication of decreased alcohol consumption after September 11th, although these effects were modest. These analyses provide little support for popular assertions that September 11th resulted in lasting and measurable impacts on Americans' well-being. PMID- 16259149 TI - Stress, life events, and socioeconomic disparities in health: results from the Americans' Changing Lives Study. AB - It has been hypothesized that exposure to stress and negative life events is related to poor health outcomes, and that differential exposure to stress plays a role in socioeconomic disparities in health. Data from three waves of the Americans' Changing Lives study (n = 3,617) were analyzed to investigate prospectively the relationship among socioeconomic indicators, five measures of stress/negative life events, and the health outcomes of mortality, functional limitations, and self-rated health. The results revealed that (1) life events and other types of stressors are clearly related to socioeconomic position; (2) a count of negative lifetime events was positively associated with mortality; (3) a higher score on a financial stress scale was predictive of severe/moderate functional limitations and fair/poor self-rated health at wave 3; and (4) a higher score on a parental stress scale was predictive of fair/poor self-rated health at wave 3. The negative effects of low income on functional limitations attenuated to insignificance when waves 1 and 2 stress/life event measures were controlled for, but other socioeconomic disparities in health change remained sizable and significant when adjusted for exposure to stressors. The results support the hypothesis that differential exposure to stress and negative life events is one of many ways in which socioeconomic inequalities in health are produced in society. PMID- 16259151 TI - Systemic delivery versus elution of drugs that enhance device performance. AB - Questions are being raised as to whether it is better to deliver a drug systemically or via a controlled-release mechanism in situations where it is intended that the drug would enhance medical device performance. The issues are addressed here in relation to cardiovascular devices and orthopaedics. PMID- 16259150 TI - Environmental stressors: the mental health impacts of living near industrial activity. AB - A growing literature examines whether the poor, the working class, and people of color are disproportionately likely to live in environmentally hazardous neighborhoods. This literature assumes that environmental characteristics such as industrial pollution and hazardous waste are detrimental to human health, an assumption that has not been well tested. Drawing upon the sociology of mental health and environmental inequality studies, we ask whether industrial activity has an impact on psychological well-being. We link individual-level survey data with data from the US. Census and the Toxic Release Inventory and find that residential proximity to industrial activity has a negative impact on mental health. This impact is both direct and mediated by individuals' perceptions of neighborhood disorder and personal powerlessness, and the impact is greater for minorities and the poor than it is for whites and wealthier individuals. These results suggest that public health officials need to take seriously the mental health impacts of living near industrial facilities. PMID- 16259152 TI - Design considerations when using semicrystalline engineering polymers. AB - Intelligently exploiting the material properties of polymers can lead to multifunctional components that are commercially and functionally superior to previous designs. This article explains how to achieve this. PMID- 16259153 TI - Measuring volumes with gas mass flow sensors. AB - With some design care, a mass flow sensor, pressure transducer and a temperature sensor can provide a fast and accurate method of measuring many difficult volumes. PMID- 16259155 TI - Understanding US labelling requirements. AB - Medical device manufacturers should not assume that the information and symbols on labels for the European market are always suitable to meet United States (US) requirements. This article discusses US labelling requirements and how they differ from European requirements. PMID- 16259154 TI - Polyurethane scaffolds for meniscal tissue regeneration. AB - More than 1 million procedures for the total or partial removal of the meniscus in the knee joint are performed in the United States and Europe each year. These meniscectomies lead to degenerative changes of the knee and to immobility of the patients. A polyurethane scaffold is described here, which has been developed as an alternative repair solution. PMID- 16259156 TI - Applying the new medical collateral EMC standard EN60601-1-2:2001. AB - Manufacturers of electromedical equipment must test their products to the new requirements set out in collateral standard EN 60601-1-2:2001. The new tests to ensure correct risk analysis are outlined here. PMID- 16259157 TI - Making product commercialisation more successful. AB - Turning an idea or concept into a winning product is one of the toughest challenges for lone innovators as well as global companies. The four critical elements of the process are discussed here. PMID- 16259158 TI - The innovation process centre stage. AB - London's Olympia 2 was the venue for the first Medical Device Technology Innovation Forum. The event was characterised by dialogue and discussion that explored the key issues involved in delivering clinical need through innovative products to the end user. Highlights of the event are reported here. PMID- 16259159 TI - Detecting surface issues. AB - Surface analysis can be a significant aid in product development. Recent advances in these techniques allow the fine details of different coatings to be compared and optimised coating processes to be developed. PMID- 16259161 TI - Cold curing ultraviolet adhesives. PMID- 16259160 TI - Plasma processing of surfaces. PMID- 16259162 TI - Novel micro-probe sensors for real-time measurement of ATP/Purines in the brain. PMID- 16259163 TI - Sourcing made easy. AB - MDT Sourcing is a free online service for buyers seeking the range of services used in the design, manufacture and marketing of finished medical devices. Take up of the service has been good since its launch earlier this year. Some of the latest requests for quotations are outlined here. PMID- 16259164 TI - Too many meetings? PMID- 16259165 TI - The epidemiology of terrorism casualties. PMID- 16259166 TI - Outcome after transthoracic resection of carcinoma of the oesophagus and oesophago-gastric junction. AB - AIMS: To assess the postoperative morbidity and mortality, length of stay and long-term survival after resection of carcinoma of the oesophagus and gastro oesophageal junction, after establishment of a new surgical team unit between thoracic and gastroenterologic surgeons. METHODS: We analysed the prospective collected data of 166 consecutive patients who underwent a transthoracic oesophageal resection between June 1997 and December 2003. RESULTS: There were 119 men and 47 women. The median age was 63 years (range 36-81). Fifty-five patients (33%) had squamous cell carcinoma and 111 (67%) had adenocarcinoma. Postoperative complications occurred in a total of 60 patients (36%). Ten patients (6%) died postoperatively, eight (4.8%) due to medical and two (1.2 %) due to surgical complications. The median postoperative length of stay was 11 days (range 6-75). The overall 3- and 5- years survival was 35.6% and 30.6% respectively. Survival was adversely affected by patient age and tumor stage. CONCLUSIONS: Concentrating resection for carcinoma of the oesophagus and oesophagogastric junction to a dedicated team of specialists, including both gastrointestinal and thoracic surgeons as well as thoracic-anaesthesiological know-how, results in acceptable complication rates as well as low mortality rates especially due to surgical complications. PMID- 16259167 TI - Intraoperative cholangiography can be safely omitted during laparoscopic cholecystectomy: a prospective study of 413 consecutive patients. AB - BACKGROUND AND AIMS: The aim of the study was to show that laparoscopic cholecystectomy (LC) can be performed safely without intraoperative cholangiography (IOC). MATERIAL AND METHODS: We conducted a prospective study of 413 consecutive patients with symptomatic gallstone disease, who underwent LC. According to the preoperative clinical, laboratory and ultrasound criteria, 38 patients (9.2%) were selected for preoperative endoscopic retrograde cholangiography (ERC). All patients were followed postoperatively for symptoms and signs of common bile duct (CBD) stones. RESULTS: Preoperative ERC allowed to make a diagnosis of choledocholithiasis in 22 (58%) of the 38 selected patients. Stone clearance was achieved with endoscopic sphincterotomy (ES) in all cases. Three patients (7.9%) had an episode of mild self-limited pancreatitis after the procedure. Eight patients (1.9%) of 413 required conversion from LC to open cholecystectomy. There were no CBD injuries and no death cases. Of the postoperative complications, 1.5% were recorded during hospital stay. During the follow-up period, for at least 2 years after surgery, retained CBD stones were verified in 6 patients (1.5%); however, the supposed rate of residual stones was 2.4%. CONCLUSIONS: This study demonstrates that performance of selective preoperative ERC with ES when necessary, followed by LC, is an appropriate and safe approach to the treatment of patients with cholecystolithiasis and unsuspected choledocholithiasis. This approach allows to omit IOC and to perform LC safely without biliary duct injuries, ensuring low rate of retained CBD stones in the late follow-up period. PMID- 16259168 TI - Acute appendicitis: the reliability of diagnosis by clinical assessment alone. AB - BACKGROUND AND AIMS: This prospective study aimed to review the trustworthiness of the initial clinical assessment in acute appendicitis without employment of imaging modalities, laparoscopy or any other adjunct diagnostic test. PATIENTS AND METHODS: 717 patients were operated on for appendicitis by six different surgeons. Initial clinical and laboratory examination were evaluated in relation to the intraoperative and the pathological appreciation of the appendiceal inflammation. RESULTS: 598 patients were found to have appendicitis, 34 a different condition, 41 had both appendicitis and an additional condition and 44 no pathology. 6% of the laparotomies and 11% of the appendectomies were unnecessary. The severity of the inflammation correlated significantly with periumbilical pain, pain migrating to right lower quadrant, loss of appetite, fever, rebound tenderness, local rigidity, polymorphonuclear predominance on deferential, polymorhonucleosis and leukocytosis. Leukocytosis was less frequent in aged patients. All the six surgeons were found to be equally reliable, although they all underestimated the setting compared to the pathologists. Patients with a long duration of symptoms had milder forms of inflammation and increased percentage of unanticipated abdominal conditions. CONCLUSIONS: Appendicitis can be reliably diagnosed clinically without employment of adjunct tests. These can be reserved for equivocal cases. PMID- 16259169 TI - Epidemiologic aspects of complete rectal prolapse. AB - BACKGROUND AND AIMS: This study was undertaken to find out the incidence of rectal prolapse. MATERIAL AND METHODS: Ninety-nine patients operated on for rectal prolapse at Jyvaskyla Central Hospital were studied. Patients operated between 1988 and 1998 were studied retrospectively from hospital records using chart review and thirty-five patients operated on between 1999 and 2002 were studied prospectively using our proctologic database. RESULTS: The annual incidence of diagnosed complete rectal prolapse in the district of Central Finland was mean 2.5 (range, 0.79-6.08) per 100 000 population. There were ten men (10 percent) and 89 women (90 percent). Median age of the patients was 69 (range, 21-91) years. Forty-eight percent of the patients had concomitant cardiovascular disease and 15 percent psychiatric illness. Anal incontinence affecting quality of life was seen in 64 percent and constipation in 72 percent of patients. Constipation tended to be more attributed to difficult evacuation (72 percent) than to impaired bowel action (18 percent). CONCLUSION: The annual incidence of rectal prolapse is 2.5 per 100 000 population. Rectal prolapse is associated with anal incontinence and constipation in majority of patients. PMID- 16259170 TI - Demand for plastic surgical operations after primary breast cancer surgery. AB - BACKGROUND AND AIMS: The aim of this study is to evaluate the demand for plastic operations after primary breast cancer surgery. In addition, this study aims to increase knowledge on factors affecting the wish of women for breast reconstruction. MATERIAL AND METHODS: A questionnaire was sent to some 111 patients who had undergone either mastectomy or breast conserving surgery. The response rate was 76%. The questionnaire consisted of 20 structured questions. The data was analysed statistically using the t test and the chi-square test. RESULTS: 28% of the mastectomized patients wanted a breast reconstruction. Factors found to affect the patient's wish for reconstruction were age (p < 0.001) and whether the patient had received radio- (p < 0.05) or chemotherapy (p < 0.05). A difference was found between the mastectomy group and the breast conserving surgery group as to satisfaction in the general (p < 0.05) and the cosmetic (p < 0.05) outcome of the surgery, as well as patient's expectations regarding the outcome (p < 0.01). CONCLUSIONS: Rather fewer mastectomized patients wanted a reconstruction. The proportion, however, grew significantly larger in the younger age groups. This study also shows that receiving radio- or chemotherapy seems to predict a woman's choice against a reconstruction. Patients who receive radio- or chemotherapy may consider their illness more severe and life-threatening than those not receiving such treatments. PMID- 16259171 TI - Risk assessment for severe postoperative hypocalcaemia after neck exploration for primary hyperparathyroidism. AB - BACKGROUND AND AIMS: A further development of the risk profile for severe postoperative hypocalcaemia after surgery for primary hyperparathyroidism (pHPT) was made with the aim of expanding the group of patients who can be discharged safely after 23 hours. METHODS: Prospective study with 156 consecutive pHPT patients (158 operations) during 2001 and 2002. Risk factors for postoperative severe hypocalcaemia (ionised calcium < 1 mmol/L), were (1) preoperative concentration of parathyroid hormone (PTH) > 35 pmol/L (five times the upper reference value, reference range 1.1 to 6.9), (2) history of previous neck surgery, (3) biopsy/excision of > 2 parathyroid glands or (4) concomitant thyroid surgery. RESULTS: The risk factors showed a sensitivity of 100 % (9/9). In 110 of the operations (70%) no risk factors were identified. Postoperative calcium levels were significantly lower after 48 operations with risk factor(s) identified, as compared to the group without risk factors (p < 0.01). Seven of 17 patients (41%) with PTH > 35 pmol/L developed severe postoperative hypocalcaemia. Two of 31 patients (6%) with PTH < 35 pmol/L in the presence of other risk factor(s) developed severe postoperative hypocalcaemia. CONCLUSION: Patients with no risk factor can safely been discharged from hospital on the first postoperative day. Patients with preoperative concentration of PTH > 35 pmol/ L (five times the upper reference value) should stay in hospital until nadir level of calcium is reached. Patients with concentration of parathyroid hormone less than 35 pmol/ L in the presence of other risk factor(s) may have an early discharge from hospital (second postoperative day) combined with outpatient measurements of calcium levels. PMID- 16259172 TI - Early elective conversion for failing EVAR. AB - BACKGROUND: The aim of the study was to assess the long term results of endovascular aneurysm repair (EVAR) of abdominal aortic aneurysms at the Helsinki University Central Hospital 1996-2004 with a special emphasis on elective conversion procedures and their outcome. METHODS: Treatment results and follow-up data of all 110 elective EVAR procedures performed in our institution were gathered prospectively and evaluated. RESULTS: Conversion to open surgery was performed in 23 (21%) of 110 EVAR patients. 30-day mortality after elective stent graft deployment was 0% as it was also on elective conversions (n = 20). Secondary elective conversions were performed due to infection in two and after failing stent-graft treatment in 12 patients. Six of the conversions were performed primarily. Three urgent conversions (3% overall) were performed: two of these patients died, corresponding to an overall mortality of 9% (2/23) in all conversions. Five-year aneurysm-related mortality after EVAR among patients with elective conversion was 0% and 19% in patients with secondary procedures other than elective conversions. CONCLUSIONS: As conversions seem to be hazardous only when performed in urgent situations, elective conversion could be an alternative treatment method in complex failing first-generation stent-grafts as it may reduce mortality associated with urgent conversions or repeated attempts to maintain graft integrity with endovascular procedures. PMID- 16259173 TI - Albumin induced hypercoagulability does not reduce blood loss in patients undergoing total hip arthroplasty. AB - BACKGROUND AND AIMS: Albumin may enhance and hydroxyethyl starch (HES) may impair haemostasis. While the effects are also dependent on haemodilution we minimized it by early structured transfusion therapy, and compared albumin and HES regarding blood loss and coagulation parameters in hip arthroplasty patients. MATERIAL AND METHODS: 101 patients undergoing primary hip arthroplasty received in random order 4% albumin (n = 48) or HES (average Mw 120 kDa/molar substitution ratio 0.7, n = 53). The administration of colloid, red blood cell (RBC), fresh frozen plasma and platetet concentrates begun after a 6-8%, 12-16%, 60% and 100% blood loss of the patient's calculated blood volume respectively. Explanatory risk factors for blood loss were modelled by regression analysis. RESULTS AND CONCLUSIONS: Administration of albumin or HES 1200 ml (500-2000 and 500-1800) [median (range) respectively] did not affect blood loss. The vWF antigen was higher in the albumin group (p = 0.04) postoperatively. Haematocrit value, platelet count, bleeding time, prothrombin time value, activated thromboplastin time, FV activity and fibrinogen concentration were comparable between the groups. Long operation time was associated with great blood loss (p < 0.001). In hip arthroplasty patients with near normal levels of haematocrit albumin enhanced coagulation without altering blood loss. PMID- 16259174 TI - Comparison of digital and conventional radiostereometric image analysis in an ankle phantom model. AB - BACKGROUND AND AIMS: Radiostereometric analysis (RSA) allows accurate three dimensional measurements of micromotion in skeletal structures. The current RSA techniques are based on the analysis of scanned plain films. This study was undertaken to compare digital filmless RSA technique to conventional scanning technique using a phantom model of the ankle mortise. MATERIAL AND METHODS: In the first experiment, the relative displacement of the markers inserted to the fibula in relation to the markers inserted to the tibia was studied by means of double examinations and the precision of DICOM images were compared to scanned images of printed radiographs. In the second experiment, the film pair of double examination was re-imported or re-scanned and self-compared in order to show merely the error related to the image processing. RESULTS: The precision of RSA using scanned images of printed radiographs was compatible to DICOM images. However, the mean error of rigid body fitting (ME) values were significantly lower in use of DICOM images compared with scanned radiographs, indicating less deformation of rigid body segments in filmless analysis. CONCLUSIONS: Precision of the RSA method was improved under the completely filmless environment. Therefore, this technique can be recommended for clinical studies of radiostereometric analysis. PMID- 16259175 TI - No long-term effects of ultrasound therapy on bioabsorbable screw-fixed lateral malleolar fracture. AB - BACKGROUND AND AIMS: The present study was initiated to evaluate the long-term effects of low-intensity ultrasound therapy on bioabsorbable screw-fixed lateral malleolar fractures, which has not been studied earlier. PATIENTS AND METHODS: The study design was prospective, randomized, double-blinded, and placebo controlled. Sixteen dislocated lateral malleolar fractures were fixed with one bioabsorbable self-reinforced poly-L-lactide screw. The patients used an ultrasound device 20 minutes daily for six weeks without knowing it was active (eight patients) or inactive (eight patients). The follow-up time was 18 months. The radiological bone morphology was assessed by multidetector computed tomography (MDCT) scans, the bone mineral density by dual-energy X-ray absorptiometry scans, and the clinical outcome by Olerud-Molander scoring and clinical examination of the ankle. RESULTS: The MDCT scans revealed that all fractures were fully healed, and no differences were observed in radiological bone morphology at the fracture site. The bone mineral density of the fractured lateral malleolus tended to increase slightly during the 18-month follow-up, the increase being symmetrical in both groups. No differences were observed in the clinical outcome or Olerud-Molander scores. CONCLUSIONS: The six-week low intensity ultrasound therapy had no effect on radiological bone morphology, bone mineral density or clinical outcome in bioabsorbable screw-fixed lateral malleolar fractures 18 months after the injury. PMID- 16259176 TI - Postoperative wound complications after internal fixation of closed calcaneal fractures: a retrospective analysis of 126 consecutive patients with 148 fractures. AB - AIM: The aim of the study was to analyse the number of soft-tissue complications after internal fixation of calcaneal fractures and to evaluate risk factors leading to these complications. MATERIAL: A retrospective analysis of 126 consecutive patients with 148 operatively treated calcaneal fractures was performed. Only primarily closed fractures were included in the study. RESULTS: Wound healing was problematic in 35 cases (24 per cent). The wound was infected in 23 cases (16 per cent) and a wound edge necrosis was observed in 12 cases (8 per cent). The soft tissue complication needed operative treatment in 20 cases (14 per cent). The complications did not lead to amputations in any case. In the end of the follow up all wounds had healed. CONCLUSIONS: The statistical analysis identified a longer delay in surgery and longer operation time to be positive risk factors for wound complications. PMID- 16259177 TI - Intestinal microdialysis--applicability, reproducibility and local tissue response in a pig model. AB - BACKGROUND AND AIMS: Microdialysis has been applied to the intestinal wall for the purpose of monitoring local ischemia. The aim of this study was to investigate the applicability, reproducibility and local response to microdialysis in the intestinal wall. MATERIALS AND METHODS: In 12 pigs two microdialysis probes were inserted into the ileal wall, one in the peritoneal cavity and one in the psoas muscle. Relative recovery was measured for all probes by the no net flux method. Metabolic measurements of glucose, lactate and glycerol were performed over six hours. The ileal wall segments containing the probes were processed for histological examination. RESULTS: Large intra- and inter-group differences in the relative recovery were found between all locations. Absolute values of metabolites showed no significant changes during the study period. The lactate in blood was 25-30% of the intra-tissue values. A severe inflammatory reaction was seen in the ileal wall around all probes. CONCLUSION: Measurement of the relative recovery is essential for valid measurements of metabolites when using microdialysis. The inflammatory reaction around the probe in the intestinal wall is likely to affect metabolism and measurements hereof. Therefore intestinal wall microdialysis seems confined to experimental research, and future studies should consider the intra-peritoneal approach. PMID- 16259183 TI - Keeping up with mobile chest drains. PMID- 16259184 TI - Do your part to stave off STREP. PMID- 16259185 TI - Radiofrequency catheter ablation. PMID- 16259186 TI - 2005 earings survey: how does your pay stack up? PMID- 16259187 TI - Hurricane Katrina: nursing in the eye of the storm. PMID- 16259188 TI - A wild reaction to a topical anesthetic. PMID- 16259189 TI - Nurse wins battle for cancer drug. PMID- 16259190 TI - Ageism must be tackled through education. PMID- 16259191 TI - Under pressure. PMID- 16259193 TI - Improving identification and awareness of hepatitis C. AB - Hepatitis C has been described as a silent epidemic, as most of those infected show no symptoms over a long period. A report from the Hepatitis C Trust (2005) raises concerns that detection and treatment is low compared to that in the rest of Europe. Improvements in screening, treatment and public awareness are needed in order to reduce potential death from liver disease related to infection. PMID- 16259192 TI - Four-star service. PMID- 16259194 TI - Mouth cancer. PMID- 16259195 TI - Blood cells. Part three--granulocytes and monocytes. PMID- 16259196 TI - Management and effects of parenteral nutrition. AB - Despite nutrition being a basic human need, malnutrition in hospitals remains surprisingly common, with the effects clearly documented (Taylor and Goodison McLaren, 1992). Malnourished patients are more likely to suffer from complications; are at increased risk of developing infections; have poor or delayed wound healing, increased mortality rates and longer hospital stays. This article focuses on the basics of parenteral nutrition in the adult, addressing a small selection of potential complications. PMID- 16259197 TI - Adhesive film dressings. PMID- 16259198 TI - Improving referral information in community mental health. AB - This article explains how a questionnaire to improve the quality of information about clients referred to a community mental health team was devised. The questionnaire was drawn up with the help of the local advocacy service and service-user representatives, and led to improved information that enabled us to improve our service to clients. PMID- 16259199 TI - The evidence on quality of life for older people. AB - Quality of life is a major concern for older people and many factors contribute to how this group perceive the concept. Quality of life relates to the individual person therefore a good quality of life means different things to different people. Health care professionals must place an emphasis upon promoting quality of life and educating older people on how it can be optimised. PMID- 16259200 TI - Welcome outcomes of the GMS contract. PMID- 16259201 TI - Changes in thte provision of home oxygen therapy. PMID- 16259203 TI - Placebo inhaler devices and infection risks. PMID- 16259202 TI - Principles for inserting and managing chest drains. PMID- 16259204 TI - Incentive spirometry for children with sickle cell disorder. PMID- 16259206 TI - Nursing, for all its strengths is anything but single-minded. PMID- 16259205 TI - Reducing the toll of Parkinson's. PMID- 16259207 TI - Mortality in a recent study of 625 patients with chronic obstructive pulmonary disease compared with results of 3 older studies. AB - OBJECTIVE: To assess comparative mortality in COPD patients by severity in a recent study with results in 3 older studies. METHOD: Analysis is made of a recent multicenter study (7 clinics in the United States, Spain, and Venezuela) of COPD patients. An evaluation cohort of 207 patients was utilized to establish a scoring system based on body mass index (B), airflow obstruction (O--measured by FEV1, forced expiratory volume at 1 second), dyspnea (D), and exercise capacity (E). A scoring system for each of these 4 severity factors led to the development of the BODE Index, with a range of 0-10. This index was shown to produce a wider range of mortality than staging (1 to 3) by the FEV1 alone. RESULTS: From the FEV1 Stage and the BODE Index data, a validation cohort of 625 COPD patients was observed for 52 months. This follow-up showed a wider range of mortality by the BODE Index than that obtained by use of FEV1 staging alone. This recent experience (1997-2002) is compared with results of 3 previously published mortality studies of COPD. CONCLUSION: Incorporation of additional severity factors such as dyspnea and exercise capacity improves the prediction of mortality by severity of the COPD, as compared with the use of FEV1 staging alone. Mortality remains at a very high level in all cases, except for those with the mildest form of COPD. PMID- 16259208 TI - The association between the length of the QT interval and mortality in the Cardiovascular Health Study: a mortality analysis. AB - Prolonged QT interval was found to be associated with increased cardiac and all cause mortality in a subgroup of the Cardiovascular Health Study (CHS). A mortality analysis is presented of this study, underscoring the need to use the appropriate expected group. PMID- 16259209 TI - Practical cancer genetics, genomics and proteomics. AB - Functional genomics is the science that defines the function of newly identified genes and translates this knowledge into related health and disease. Functional genomics is complex, involving multiple scientific disciplines including computational biology, genetics, physiology, structural biology, and molecular and cell biology. Due to its inherent complexity, the National Institutes of Health (NIH) has termed functional genomics "the science of collaboration." This paper will review microarray gene expression analysis, proteomics, and the impact of molecular medical genetics on medical care. PMID- 16259210 TI - Update on HIV infection. AB - In the third decade of the global epidemic, it is evident that human immunodeficiency virus (HIV) disease is quite different from the disease first recognized among a small number of homosexual men in 1981. The spread of HIV has been particularly alarming in developing countries, especially sub-Saharan Africa and Southeast Asia, and it continues to threaten other populations in Eastern Europe, Latin America and the Caribbean. HIV therapeutic advances have resulted in a marked decrease in acquired immunodeficiency syndrome (AIDS) incidence and death in the United States and Western Europe. With the introduction of "triple therapy," antiretroviral treatment has resulted in an extraordinary increase in the quality of life and life expectancy among HIV-infected persons. However, the rate of decline in AIDS incidence and deaths from the latter part of 1998 through 2000 has slowed for a number of reasons. Even with the stabilization of HIV in Western countries, it remains as important as ever to follow sound insurance principles to address HIV risk. The clinical success stories have yet to be translated into the insurance realm. A very strict analysis of the medical literature will be needed. PMID- 16259211 TI - Endovascular treatment of carotid artery stenosis. AB - Carotid artery stenting (CAS) was originally developed as an alternative to carotid endarterectomy (CEA) for high-risk patients. CAS is illustrated with before and after angiograms showing severe recurrent stenosis treated with a self expanding stent. Indications and available outcome data for CEA and CAS are summarized. The future for CAS may well be bright, but the procedure is early in its development. Further trials are needed to demonstrate its long-term efficacy compared with CEA. PMID- 16259212 TI - Left axis deviation. AB - Left axis deviation is one of the most commonly encountered ECG abnormalities. Its presence should alert medical directors and underwriters to the possibility of underlying structural heart disease. Many of the causes of left axis deviation are apparent from the clinical findings. Left anterior fascicular block is one of the commonest causes of left axis deviation and has specific ECG criteria for its diagnosis. PMID- 16259213 TI - A case of cutaneous lymphoma. AB - Cutaneous lymphomas are relatively rare neoplasms, yet they are occasionally encountered in underwriting. The classification of cutaneous lymphomas leaves many in a quandary. The following case represents an uncommon cutaneous lymphoma. It will be used as a springboard to present a classification of cutaneous lymphomas, the staging definitions, prognostic variables, and some mortality data. PMID- 16259214 TI - Arterial dissection as a cause of stroke. PMID- 16259215 TI - Arrhythmogenic right ventricular dysplasia. AB - Arrhythmogenic right ventricular dysplasia is a type of nonischemic cardiomyopathy that is characterized by fibrofatty replacement of the right ventricle and is an important cause of ventricular arrhythmias in younger people. PMID- 16259216 TI - On a historical note: the legacy of Thomas Hodgkin. AB - Physicians in the practice of insurance medicine are exposed to an unbelievable spectrum of pathology. This series of short biographies is intended to give the reader a quick overview of the history of diseases encountered in our everyday practice and to see "the face behind the name." In the first article of this series, I wish to pay homage to one of the "great men of Guy's," Dr. Thomas Hodgkin. PMID- 16259217 TI - Understanding polycystic ovarian syndrome. PMID- 16259218 TI - JNK: bridging the insulin signaling and inflammatory pathway. AB - Obesity and insulin resistance are strongly associated with systemic markers of inflammation and endoplasmic reticulum stress. c-Jun N-terminal kinases (JNK) are activated by inflammatory cytokines and have a key role in beta-cell apoptosis and in negative regulation of insulin signaling. JNK1-deficient mice are protected from diet-induced obesity and insulin resistance, while genetically obese mice with targeted mutations in JNK1 are leaner and have reduced insulin and blood glucose levels. These studies validate JNK as a link between inflammation and metabolic diseases and as a promising drug target. This review highlights recent advances in small-molecule inhibitors of JNK that have also been targeted for other diseases with an inflammatory component such as stroke, rheumatoid arthritis, and Alzheimer's and Parkinson's diseases. PMID- 16259219 TI - Adiponectin and its potential in the treatment of obesity, diabetes and insulin resistance. AB - Adiponectin is a protein hormone produced exclusively by adipocytes. Its circulating levels are decreased in individuals with obesity, atherosclerosis and insulin resistance, suggesting that its deficiency may have a causal role in the etiopathogenesis of these diseases. Studies have shown that adiponectin administration in rodents has insulin-sensitizing, anti-atherogenic and anti inflammatory effects and under certain settings also decreases body weight. Therefore, adiponectin replacement in humans may represent a promising approach to prevent and/or treat obesity, insulin resistance and type 2 diabetes; however, clinical studies with adiponectin administration need to be conducted to confirm this hypothesis. Current experimental and clinical data regarding adiponectin physiology and pathophysiology are detailed in this review. PMID- 16259220 TI - Nuclear receptor regulation of lipid metabolism: potential therapeutics for dyslipidemia, diabetes, and chronic heart and liver diseases. AB - Lipids are essential components of biological membranes, fuel molecules and metabolic regulators that control cellular functions, metabolism and homeostasis. The liver plays a central role in regulating lipid metabolism and whole body lipid homeostasis. Sterols, bile acids and fatty acids are the endogenous ligands of the liver orphan receptor, farnesoid X receptor, peroxisome proliferator activated receptor, vitamin D receptor, constitutive androstane receptor and pregnane X receptor. These metabolic receptors coordinately regulate lipid, glucose, energy and drug metabolism. Alteration of lipid homeostasis causes dyslipidemia, which is a major risk factor contributing to atherosclerotic cardiovascular diseases, diabetes, obesity and liver diseases. Advances in the understanding of the mechanisms of nuclear receptor regulation of lipid homeostasis have provided an opportunity to investigate potential therapeutic drugs targeted to nuclear receptors. This could be useful for the treatment of diabetes, and cardiovascular and chronic liver diseases. PMID- 16259221 TI - Neuropeptide Y2 receptors as drug targets for the central regulation of body weight. AB - During the past decade, a detailed understanding has emerged of the aminergic and peptidergic neural pathways present within the brain that regulate appetite. Central among the peptide regulators is neuropeptide Y (NPY), a potent orexigenic agent that acts through five different receptor subtypes. Efforts to find novel appetite suppressant drugs that inhibit the interaction of NPY with either the NPY Y1 or NPY Y5 receptor subtypes have proven disappointing. Attempts have now been made to identify an NPY Y2 stimulator that will suppress appetite. Within the hypothalamus, NPY Y2 receptors have a predominantly presynaptic location where they act to inhibit NPY release. Stimulation of NPY Y2 receptors with synthetic peptide ligands or the gut derived peptide PY3-36 has been shown to reduce food intake. The NPY Y2 receptor has a wide distribution both within the brain and in the periphery. Stimulation of the NPY Y2 subtype at these sites produces a wide array of effects unrelated to changes in food intake. In consequence, the administration of both endogenous and exogenous agonists of the NPY Y2 receptor is likely to cause side effects, particularly regarding pituitary hormone release, as well as on the cardiovascular and gastrointestinal systems. The possibility that long-term NPY Y2 agonism could cause bone thinning and retinal angiogenesis are of particular concern and will need to be investigated as drug discovery moves forward. PMID- 16259222 TI - Use of insulin sensitizers in polycystic ovarian syndrome. AB - Polycystic ovarian syndrome is a common disorder associated with a significant long-term risk of developing type 2 diabetes and cardiovascular diseases. Insulin resistance and hyperinsulinemia play an important role in its pathophysiology and therefore insulin sensitizers have been proposed as a possible treatment option for this condition. In this review, pertinent literature is described that supports the use of insulin sensitizers for the management of short-term (fertility and hyperandrogenism) as well as long-term (type 2 diabetes, cardiovascular diseases and endometrial cancer) clinical issues of the syndrome. There is sufficient evidence in the literature to support the initial use of insulin sensitizers for fertility and the chronic treatment of hyperandrogenism. Furthermore, insulin sensitizers may prevent type 2 diabetes or cardiovascular diseases, whereas some evidence suggests that oral contraceptives could increase these risks. Therefore, although oral contraceptives may provide a more reliable control of menstrual disorders, insulin sensitizers should be considered as a preferential treatment option in women with polycystic ovarian syndrome at an increased risk of developing type 2 diabetes or cardiovascular disease, especially if they do not need contraception. PMID- 16259223 TI - Pharmacological options in the treatment of acromegaly. AB - The last decade has seen two major changes in the management of patients with growth hormone (GH)-secreting pituitary tumors. Firstly, evidence has revealed a common pattern of increased mortality in patients with elevated GH and/or the target growth factor insulin-like growth factor-I (IGF-I). Secondly, the availability of highly specific and selective pharmacological agents to lower GH and/or IGF-I levels has greatly facilitated a more aggressive approach to the treatment of patients with persistently active acromegaly. This review will summarize the current major classes of pharmacotherapeutic agents with special emphasis on their recognized benefits and risks. PMID- 16259225 TI - E1-INT (Transition Therapeutics/Novo Nordisk). AB - Transition Therapeutics (through its acquisition of Waratah Pharmaceuticals), in collaboration with Novo Nordisk, is developing E1-INT, an injectable islet neogenesis therapy comprising an epidermal growth factor analog and a gastrin analog, for the treatment of insulin-dependent (type 1) and non-insulin-dependent (type 2) diabetes. The compound is currently undergoing phase II clinical trials. PMID- 16259224 TI - Androgen replacement therapy in women. AB - Physiologically, androgens in women either derive from direct ovarian production or from the peripheral conversion of the adrenal sex steroid precursor dehydroepiandrosterone to the active androgens. Therefore, loss of adrenal or ovarian function, as in Addison's disease or after bilateral oophorectomy, usually results in severe androgen deficiency. Androgen replacement in these women may produce significant improvements, particularly in libido and mood. Physiological menopause is not necessarily associated with androgen deficiency and therefore does not routinely require androgen therapy. The number of randomized controlled trials of androgen use in women is still limited. Choosing both a convenient and efficient mode of androgen administration in women remains a challenge and currently none of the available preparations is officially approved for use in women. PMID- 16259226 TI - DiaPep277 (DeveloGen). AB - DiaPep277, a 24-amino acid peptide based on residues 437 to 460 of heat shock protein 60, is undergoing phase II clinical trials by DeveloGen for the potential treatment and prevention of established and newly diagnosed type 1 diabetes symptoms of the prediabetic state and of latent autoimmune diabetes of the adult. PMID- 16259227 TI - APD-356 (Arena). AB - Arena is developing APD-356, the lead in a series of orally active, small molecule 5-hydroxytryptamine 2C agonists for the potential treatment of obesity and diabetes. A phase IIb trial was initiated in June 2005, and preliminary results were expected at the end of 2005. PMID- 16259228 TI - PREOS NPS (Allelix/Nycomed). AB - NPS Allelix and Nycomed are developing PREOS, an injectable recombinant human parathyroid hormone, for the potential treatment of osteoporosis. In May 2005, NPS filed a market authorization application for PREOS in the US which was accepted for review in July 2005. PMID- 16259229 TI - Lasofoxifene (Pfizer). AB - Pfizer is developing lasofoxifene, an oral naphthalene derivative selective estrogen receptor modulator identified under a collaboration with Ligand, for the treatment of osteoporosis and vaginal atrophy. In August 2004, Pfizer submitted a New Drug Application to the Food and Drug Administration (FDA) seeking approval of lasofoxifene for the treatment of osteoporosis; however, in September 2005, the FDA issued a non-approvable letter. PMID- 16259230 TI - Mary Glover Lecture 2004: leaving a legacy. AB - Mary Glover was a Head Nurse at St. Paul's Hospital in Vancouver. She was killed in a plane crash more than 25 years ago. Yet, through this neuroscience nurse's passion for her specialty, we share in her legacy through the annual Mary Glover Lecture, which was established by her parents after her death. The first Mary Glover Lecturer was Pamela Mitchell, a well-known neuroscience nurse from the School of Nursing at the University of Washington. She is leaving a multifaceted legacy through her research on intracranial pressure and quality of care as well as her books and her mentorship. Jessie Young has left a legacy as the founder and first president of the Canadian Association of Neuroscience Nurses (CANN). CANN is leaving a legacy with many firsts among Canadian nursing specialty organizations. Leaving a legacy is not just about donating money or writing a famous book. For most of us, our legacy comes in the little everyday things of life. Ask yourself, what is the legacy that you are leaving as a neuroscience nurse and as an individual? PMID- 16259231 TI - Psychosocial aspects of caregiving to stroke patients. AB - A high percentage of individuals who have suffered a stroke will be cared for at home, primarily by aging spouses and/or relatives. Providing care to a family member with a chronic or life-threatening condition can be both emotionally rewarding and distressing for the care provider. The objective of this research was to test the factors associated with caregiver experiences. The findings of a convenience sample of 48 caregivers indicated that the higher the amount of caregiver burden, the greater the lifestyle impact and emotional distress for the caregiver. Caregiver satisfaction was not found to be associated with emotional well-being. The amount of support, both instrumental and social, did not improve the emotional well-being of the caregiver. The caregiver's sense of mastery was found to moderate the relationship between lifestyle impact and emotional well being and also between caregiver satisfaction and emotional well-being. PMID- 16259232 TI - Post-stroke shoulder subluxation: a concern for neuroscience nurses. AB - Approximately 84% of all stroke patients with hemiplegia will experience shoulder injury and pain. The importance of maintaining proper posture while positioning and transferring a stroke patient is key to decreasing risk for shoulder injury. Shoulder subluxation injury post-stroke is a consequence of sustained hemiplegia and spasticity. Current research evidence suggests that using therapies such as gentle range of motion and functional electrical stimulation may reduce and prevent shoulder subluxation and hemiplegic shoulder pain. However, physiotherapists are currently the only professionals who can implement such therapies. Considering that stroke care provided by neuroscience nurses includes transferring, positioning and assisting in activities of daily living, it is clear that nurses are an important part of the therapy process. Therefore, the question is: "What is the role of the neuroscience nurse in the reduction and prevention of shoulder pain post-stroke?" The purposes of this paper are to i) discuss the causes of shoulder subluxation and related pain post-stroke, ii) review current best practice in prevention and treatment of shoulder subluxation, and iii) explore ways in which the acute neuroscience nurse can prevent or reduce shoulder subluxation in the hemiplegic stroke patient. PMID- 16259233 TI - Different strokes... PMID- 16259234 TI - Staff support after trauma in A&E. PMID- 16259235 TI - Practice makes perfect: fishing hook removal. PMID- 16259236 TI - Managing children with croup in emergency departments. PMID- 16259237 TI - Compression bandaging for soft tissue injury of the ankle: a literature review. PMID- 16259238 TI - Recognising and treating Colles' type fractures in emergency care settings. AB - In this article, the author explains how to identify a Colles' type fracture, explains how to insert a haematoma block, and gives a step by step guide to reducing this type of fracture. PMID- 16259239 TI - Board's eye view. PMID- 16259240 TI - Promina-Aetna US healthcare physician provider contracts raise troubling questions. PMID- 16259242 TI - The ultimate recycling. PMID- 16259243 TI - The men's health issue, January 1998: questions regarding the diagnosis of prostate cancer. PMID- 16259245 TI - The state of internal medicine. PMID- 16259244 TI - The men's health issue, January 1998: questions regarding the diagnosis of prostate cancer. PMID- 16259246 TI - GAFP celebrates 50 years. PMID- 16259247 TI - Organ procurement organizations: A history and overview of LifeLink. PMID- 16259248 TI - Organ donation: tragedy transformed to triumph. PMID- 16259249 TI - The role of the hospital and the OPO in the donation process. PMID- 16259250 TI - The role of immunosuppressive drugs in transplantation. PMID- 16259251 TI - The world of transplantation medicine: a bittersweet mixture. PMID- 16259252 TI - Renal failure and donation for transplantation in the African-American community. PMID- 16259253 TI - Lung transplantation: a current perspective. AB - Lung transplantation is a viable therapeutic option for patients with end-stage lung disease. Quality of life and survival are improved for most recipients. Donor availability remains an impediment to widespread application. The development of OB after lung transplantation continues to affect long-term survival. Clinical and basic science research will provide new strategies to further improve results. PMID- 16259254 TI - Living-related kidney donation. AB - Living renal donation is an effective means to expand the organ donor pool and allow more transplants to be performed at greater convenience for the recipient and with greater long-term success. Risks to the recipient appear to be minimal as long as careful donor screening is employed. Living donation has been implemented in extra-renal transplantation more recently. While the experience is limited so far, living donation is likely to remain an important option for liver and lung recipients. PMID- 16259255 TI - Increasing the availability of organs for transplantation: an ethical analysis. PMID- 16259256 TI - The recipient process. PMID- 16259257 TI - A donor family's story. PMID- 16259258 TI - A recipient's story. PMID- 16259259 TI - Brain death criteria. PMID- 16259260 TI - Surgical innovations in the treatment of children with end-stage liver disease. PMID- 16259261 TI - Allograft tissue--an unsung transplant hero. PMID- 16259262 TI - Heart transplantation--current perspectives. PMID- 16259263 TI - Bone marrow transplantation: cancer therapy comes of age. PMID- 16259264 TI - The Georgia Eye Bank and corneal transplantation. PMID- 16259265 TI - Chlamydial infections among family planning clinic patients in Georgia: who should be screened? PMID- 16259266 TI - Conservative art. PMID- 16259267 TI - The role of Georgia physicians in control of farmers' skin cancer. PMID- 16259268 TI - Legal perspectives in organ and tissue procurement. PMID- 16259269 TI - The new role of physicians under the Family and Medical Leave Act. PMID- 16259270 TI - Though challenged by shortage, OR leaders find ways to cope. PMID- 16259271 TI - OR incentive programs reward staff for ingenuity, taking initiative. PMID- 16259272 TI - National surgical QI project rolls out. PMID- 16259274 TI - Fighting fatigue for perioperative staff. PMID- 16259273 TI - Surgical robot assists in human procedure. PMID- 16259275 TI - OR team helps to win Baldrige award. PMID- 16259276 TI - Taking supply management to next level. PMID- 16259277 TI - [Cardiac tumors]. AB - Primary cardiac tumors are rare and their subdivision often difficult because of their unknown origin. In the most recent classification, cardiac tumors are divided into benign (about 75% and malignant neoplasms in relationship to their tissue differentiation (rhabdomyoma, haemangioma, etc.) or uncertain aetiology (myxoma, papillary fibroelastoma). Primary malignant tumors are maimly represented by sarcomas. The most frequent tumor is cardiac myxoma, which by itself represents about 50% of all primary cardiac neoplasms. Although non invasive technologies as trans-esophageal ecocardiography allow the detection and exact localization of cardiac mass, clinical diagnosis is often tardive. This is due, besides the intrinsic rarity, to two main factors: first, the tumor is often asymptomatic (incidental autopic finding) or; alternatively, it may show aspecif symptoms mimicking heart failure or other pathologies. In this article, clinicopathological features of main primary cardiac tumors are presented. Investigation of the histogenesis of some of these neoplasms is still a primary field of research. PMID- 16259279 TI - Malignant granular cell tumor of the abdominal wall. AB - The granular cell tumor is an uncommon tumor that usually appears as a solitary small nodular growth and runs a benign course. It occurs widely throughout the body, but is rarely described in the abdominal wall. The authors report a case of malignant granular cell tumor which was arising in anterior abdominal wall of a 67-year-old woman. Malignant variant is rare and the abdominal wall site is extremely uncommon. Regarding this clinical case and the literature the authors purpose to review the criteria of malignancy. PMID- 16259278 TI - [Splenic hamartoma]. AB - The Authors present a case of Splenic Hamartoma. This is a rare unique complex vascular lesion of the spleen. This is also a recently reinterpreted lesion with some persistent confusion regarding its definition, histogenesis and classification. The patient is a young woman who following a check up with a raised erythrocyte sedimentation rate underwent abdominal sonography demonstrating an incidental slightly hypoechoic nodular splenic lesion. The pathologic study of the splenectomy specimen showed a large (10 cm) sharply demarcated mass. Histologically the lesion presented a remarkably angiomatoid lobular-nodular configuration with abundant fibrosclerotic stroma with areas of calcification. The immunohistochemical study revealed within the angiomatoid tissue different types of blood vessels: a) capillaries CD31+, CD34+, CD8-; b) structures consistent with splenic venous sinuses CD31+, CD34+/- , CD8-/+; c) small veins CD31+, CD34+, CD8-. The Authors judge this complex picture as indicative of a Splenic Hamartoma with a peculiar lobular-nodular pattern that seems to coincide with the recently described SANT: Sclerosing Angiomatoid Nodular Transformation of the spleen. In this report the Authors discuss the pathology of the lesion and the problems concerning its vasular profile. It is also emphasized the Hamartoma's great variety of morphologic patterns derived from the preponderant growth of one or another of several histologic components. This is the cause of the presence in literature of some different pathologic entities which today are fairly recognized as part of a large pathologic spectrum of the same lesion. The Authors discuss the differential diagnosis of Splenic Hamartoma with other lesions as haemangiomas and inflammatory pseudotumor. PMID- 16259280 TI - ["Proximal type" epithelioid sarcoma of the vulva: differential diagnosis with other extrarenal rhabdoid tumors]. AB - Proximal type epithelioid sarcoma is a rare neoplasia in which morphological findings are characterized by nodular proliferation of epithelioid cells with focal rhabdoid features. It shares some histological features with other neoplasias and this gives an account of several differential diagnosis with other extrarenal rhabdoid tumors. Immunohistochemical and ultrastructural analysis are important in defining this entity: vimentin, cytokeratin, EMA and often CD34 expression of tumoral cells, moreover ultrastructurally evidence of large paranuclear whorls of intermediate filaments, are requested for diagnosis. A correct diagnostic framing is necessary because of the aggressive clinical behaviour of this tumor, that has a tendency to early spreading. We describe a case of vulvar proximal type epithelioid sarcoma in a 34 years old woman. PMID- 16259281 TI - [Cutaneous angiomyolipoma: a case report and literature review]. AB - Angiomyolipoma is a lesion usually observed in the kidney of patients with tuberous sclerosis. Extrarenal sites are very unusual with sporadic cases in internal organ, soft tissue and skin (fifteen cases have been described in this site). Herein we describe an adding case located on the ear in 58-year-old man reviewing the pertinent literature. The main diagnostic differential criteria are also discussed. PMID- 16259282 TI - Tumoral, quasitumoral and pseudotumoral lesions of the superficial and somatic soft tissue: new entities and new variants of old entities recorded during the last 25 years. Part VIII: excerpta VI. PMID- 16259283 TI - [New diagnostic markers: podoplanin-d2-40]. PMID- 16259284 TI - [Increase of obesity in school children who attend ambulatory services of the Pediatric Hospital of the Sonora State]. PMID- 16259285 TI - [Myopia in school children of the marginal area of the State of Mexico]. PMID- 16259286 TI - [Effect of dietary calcium vs calcium citrate on conventional biochemical markers in perimenopausal women]. AB - OBJECTIVE: To compare the effect of calcium citrate and a calcium enriched diet on conventional biochemical markers. MATERIAL AND METHODS: Eighty-two women aged 30 to 35 years were randomized to any of three groups:A control group of 23 women who remained intact in their dietary habits and physical activity; a second group of 28 women who received 1000 mg of dietary calcium plus physical activity 30 minutes three times per week; and a third group of 31 women who received 600 mg of calcium citrate plus 500 mg of dietary calcium and physical activity three times per week for seven months. Calcaneum bone densitometry was measured to classify women into normal and osteopenic groups. Biochemical markers were measured at baseline and at the end of the study, as follows: serum alkaline phosphatase, magnesium, calcium and phosphorus, as well as the calcium/creatinine ratio in urine. RESULTS: Thirty-four percent of women were osteopenic. These women showed a significant reduction in the final level of calcium in the third group, as compared to the second group (p < 0.05, 7.4 mg/dl vs 8.8 mg/dl). The final measurement showed significant hypermagnesemia in the second group, as compared to the third group (p < 0.05). Phosphorus levels decreased in the second group (3.5 to 3.2 mg/dl) (p > 0.05). The calcium/creatinine ratio was normal in all groups. CONCLUSIONS: The second group showed a higher bone production than the third group. No group showed bone resorption. PMID- 16259287 TI - Obesity and overweight in IMSS female workers in Mexico City. AB - OBJECTIVE: To estimate the prevalence and risk factors for overweight (OW) and obesity (OB) in women working at the Mexican Social Security Institute (IMSS, per its abbreviation in Spanish) in Mexico City, using two different classification criteria. MATERIAL AND METHODS: A cross-sectional study was performed from July 1999 to September 2000. It included 588 women 20 to 65 years of age and who were working at the IMSS. The criteria used to estimate the prevalence of OW and OB were the WHO criteria and the Mexican Official Norm (NOM) for the integrated management of obesity in Mexico. RESULTS: The frequency of OB, according to WHO criteria, was 27.6% (26% adjusted) and for OW 43.2% (40.2% adjusted). According to the NOM, the levels of OB in those with short height (< 1.50 m) increased to 75% and to 52.2% for those with height > 1.50 m. Comparison of BMI between the two height groups showed no differences. The risk factor associated with OB and OW was age. Education and exercise were protective factors. CONCLUSIONS: The high prevalences detected in this particular working group highlight the importance of prevention and control of OB in health personnel because it can result in a high number of disability and premature retirement due to disease. The use of the NOM criteria for the identification of women at risk may be useful for early detection of high-risk groups. PMID- 16259288 TI - [Prevalence of Streptococcus pneumoniae serotypes on nasopharyngeal colonization in children of Mexico City]. AB - OBJECTIVE: To determine the frequency, serotypes and susceptibility profiles to eight antimicrobials in Streptococcus pneumoniae nasopharyngeal isolates from a representative sample of children under 5 years of age, residents of Mexico City. PATIENTS AND METHODS: A cross-sectional survey was conducted in 573 children aged 2 months to 5 years.A nasopharyngeal sample was taken. S. pneumoniae identification, capsular serotyping and antimicrobial susceptibility to eight antimicrobials were performed according to standardized methods. RESULTS: S. pneumoniae was isolated in 122/573 (21.4%) children. The most frequent capsular serotypes were 23F, 35, 19F, 11A and 15A; 46% of isolates corresponded to serotypes not included in the heptavalent vaccine. Low penicillin susceptibility was found in 12% of strains with 3% of highly resistant penicillin strains; erythromycin resistance was >30% and trimethoprim-sulfamethoxazole resistance >40%. None of the isolates was resistant to vancomycin, cefotaxime, amoxacillin clavulanate, chloramphenicol or ampicillin. CONCLUSIONS: Several S. pneumoniae serotypes from nasopharyngeal carriers are not included in the heptavalent vaccine. Resistance to trimethoprim-sulfamethoxazole and macrolides is high. The medical community should be aware of these results. PMID- 16259289 TI - [Efficacy of a rapid test to diagnose Plasmodium vivax in symptomatic patients of Chiapas, Mexico]. AB - OBJECTIVE: To evaluate, under laboratory conditions, the sensitivity and specificity of a rapid diagnostic test (OptiMAL), based on immunoreactive strips, to detect Plasmodium vivax infection in febrile patients in Southern Chiapas, Mexico. MATERIAL AND METHODS: The presence of parasites in blood samples of 893 patients was investigated by Giemsa-stained thick blood smear microscopic examination (gold standard). A blood drop from the same sample was smeared on immunoreactive strips to investigate the presence of the parasite pLDH. Discordant results were resolved by PCR amplification of the parasite's 18S SSU rRNA, to discard infection. RESULTS: OptiMAL had an overall sensitivity of 93.3% and its specificity was 99.5%. Its positive and negative predictive values were 96.5% and 98.9%, respectively. Signal intensity in OptiMAL strips correlated well with the parasitemia density in the blood samples (r = 0.601, p = 0.0001). CONCLUSION: This rapid test had acceptable sensitivity and specificity to detect P. vivax under laboratory conditions and could be useful for malaria diagnosis in field operations in Mexico. PMID- 16259291 TI - ["Damage, not age, is what makes a person old": ageing and health in rural males]. AB - OBJECTIVE: To analyze the social factors associated with the health status of elderly men, 60 years and older, who live in poverty in the rural areas of Mexico. MATERIAL AND METHODS: A cross-sectional, descriptive study was conducted between February and October 2002, in a convenience sample of 392 elderly men residing in rural areas of the states of Guerrero, Morelos and Jalisco states. A validated questionnaire was applied to collect data on sociodemographic characteristics, general wellbeing, health problems, food consumption, and some indicators of mental health, personal strength and social support. Data analysis was performed with the SPSS v. 11. software, to obtain chi-squared tests; food intake measures were analized with one-way ANOVA. RESULTS: Study subjects were married or were widowers and had six children on average. A higher proportion of the elderly men of Morelos state reported to have suffered more illnesses and a higher consumption of alcohol when compared to the other two state groups. Sixty percent of the men in the study used both home remedies and medication for their health problems. More than one half of them did not receive adequate social support from their families. CONCLUSIONS: The results of this study suggest that the process of ageing among rural elderly men is perceived as a burden, mainly because of the context of poverty in which they live. The lack of appropriate income and access to health services through adequate pension and retirement plans for the elderly poor is a problem that requires immediate attention. PMID- 16259290 TI - [Trend of antimicrobial drug-susceptibility of blood isolates at an oncological center (1998-2003)]. AB - OBJECTIVE: To describe the patterns of antimicrobial resistance organisms isolated in blood cultures from patients detected in a tertiary level of care, teaching oncological hospital. MATERIAL AND METHODS: All strains obtained from blood cultures from 1998 to 2003 were included and processed using the Bactec and Microscan system to determinate isolates and susceptibility to antimicrobials. The percent difference (increase or decrease) was obtained by comparing the frequency of resistance at baseline and at the end of the study. RESULTS: A total of 2071 positive blood cultures were obtained; 59.7% of isolates were Gram negative bacteria, 35.7% Gram-positive bacteria and 4.6% were yeasts. E. coli was the most frequent isolated (18.6%), followed by. Staphylococcus epidermidis (12.7%) and Klebsiella spp (9%). Throughout the study the susceptibility of Gram negative bacteria was stable and over 88% for most of the antimicrobials tested (except for Pseudomonas aeruginosa). Ciprofloxacin susceptibility for Escherichia coli stayed around 50%. Susceptibility to amikacin was higher than that to gentamicin. Staphylococcus aureus susceptibility for oxacillin was 96% and that for vancomycin 100%. S. epidermidis susceptibility for oxacillin was 14% and for vancomycin was 98.6%. No strains of vancomycin-resistant enterococci were found. All Streptococcus pneumoniae strains were penicillin susceptible. CONCLUSIONS: The drug-resistance found in this hospital is the result of the control in the use of antimicrobials, the hospital nosocomial infection program and the use of drug combination in all patients with bacteremia. PMID- 16259292 TI - Informing the TB suspect for sputum sample collection and communicating laboratory results in Nicaragua: a neglected process in tuberculosis case finding. AB - OBJECTIVE: To assess the patient information process before sputum sample collection, the quality of sputum sample and transmission of acid-fast bacilli (AFB) examination results to TB suspects, in three local areas of Nicaragua. METHODS: (a) directed interviews of consecutive series of TB suspects whose sputum had been examined for AFB; (b) directed interview of health personnel; and (c) assessment of the sputum sample quality. RESULTS: A total of 115 TB suspects and 33 health personnel were interviewed and 625 sputum samples were assessed. Results show multiple weaknesses in the process of information to the patient during sputum collections, as well as in the communication of results. CONCLUSIONS: This study unveiled an aspect usually overlooked of case finding, that is, the information process during sputum production, sputum sample quality, and the communication of results to the TB suspects. The results illustrate the need for routine assessment of the whole diagnostic process. PMID- 16259293 TI - [Eating disorders as risk factors for osteoporosis]. AB - Eating disorders (TCA per its abbreviation in Spanish) are common in young women, with an estimated prevalence of 4-5%. One of the physical complications of eating disorders, especially anorexia nervosa (AN) and eating disorder not otherwise specified (TANE) is bone mass loss, which affects both cortical and trabecular bone. The synergistic effect of malnutrition and estrogen deficiency produces significant bone mass loss, resulting from the uncoupling of bone turnover characterized by a decrease in osteoblastic bone formation and an increase in osteclastic bone resorption. The mechanisms implied in the pathogenesis of bone loss are the hypoestrogenism, hypercortisolism, serum leptin levels and insulin like growth factor decrease. Severity of bone loss in anorexia nervosa varies depending on duration of illness, the minimal weight ever and sedentarism or strenuous exercise. Long term consequences occur, such as a fracture risk increase in patients who have suffered anorexia nervosa, compared with the general population. The first treatment line to recover bone mass is nutritional rehabilitation together with weight gain. Hormonal replacement therapy may be effective if combined with an anabolic method. Osteopenia and osteoporosis are terms adopted to define the deficiency of bone mass in adults. Authors have used these terms to define densitometric data in young subjects who have not reached their peak bone mass. We suggest the term "hypo-osteogenesia" to define the deficiency in the development of bone mass in adolescents or children. PMID- 16259295 TI - [Modern medical science and Military Medical Academy]. AB - The article presents the information about the main directions of scientific investigations of Military Medical Academy and their results during the period of 1999-2000. The scientific work was conducted in conformity with demands of orders and directives of RF Ministry of Defense. 12 integrated scientific problems were formed in the annual plans of the Academy's research work. Together with traditional directions the new ones connected with the experience of troops medical support during the armed conflicts, liquidation of consequences of extreme situations, participation of military contingents in peace-making operations were developed. The complex clinical investigations of specific features of combat pathology due to firearms used by the enemy during the military operations in Afghanistan and in the Northern Caucasus are going on. In the most of clinical departments the problems of etiology, pathogenesis and treatment of servicemen' diseases under peacetime conditions are the main directions of scientific investigations. Every year the Academy's rationalizers and inventors produce 60-70 inventions and more than 500 rationalization proposals. Since 1995 the Academy publishes the journal "Clinical medicine and pathophysiology" and since 1999--"Bulletin of Russian Military Medical Academy". The Academy's scientific potential comprises 194 professors, 295 associate professors, 349 Doctors and 894 Candidates of Science, 20 Honoured Scientists of RF, 57 members and corresponding members of academies (Russian Academy of Medical Sciences, Russian Academy of Natural Sciences and other social academies). PMID- 16259294 TI - [Main Military Medical Headquarters is two centuries]. AB - Basing on the wide spectrum of sources the authors describe the history of creation, formation and functioning of the military medicine central headquarters in Russia, i.e. the Main Military Medical Headquarters. Control of military and civil medicine in Russia had a rich history. In XVI-XVII centuries there was a Pharmaceutical order in the system of State apparatus which served as the highest headquarters of health service in the country till 1707. Subsequently the problems of military and civil medicine were in charge of Pharmaceutical office (since 1707), Medical board (since 1721), Medical office (since 1725), Doctors' assembly (since 1730), Medical office (since 1732), Medical board (since 1763) and the 3rd expedition of Ministry of Home Affairs (1803-1805). In XIX the Medical expedition solved all the problems connected with control of military medical service. Later on this work was conducted by the Medical department of Military Ministry and since 1867 it was in charge of the Main Military Medical Headquarters. The experience of wars and armed conflicts confirms the necessity of strategic planning and development of modern military health service which is successfully coordinated by the main headquarters of Russian Armed Forces medical service during two centuries. Today the RF MD Main Military Medical Headquarters is the central organ in the system of RF Ministry of Defense, which heads the whole medical service of the Armed Forces of our country. PMID- 16259296 TI - [Medical service of Russian Federal Security Service]. PMID- 16259297 TI - [From the history of Frontier Forces' military medicine]. PMID- 16259298 TI - [Medical service of internal troops of Russian Ministry for Internal Affairs]. PMID- 16259299 TI - [Formation and development of Central Headquarter of the Navy medical service]. PMID- 16259300 TI - [V. S. Kudrin the chief medical inspector of Russian fleet]. PMID- 16259301 TI - [From the history of the Kronshtadt Naval Medical Library]. PMID- 16259302 TI - [The Voroshilov Central Soci sanatorium (the little-known pages of military medicine history)]. PMID- 16259304 TI - [Significance of the XXXVI World Congress Devoted to the Military Medicine]. PMID- 16259303 TI - [The first military medical museum of Russia]. PMID- 16259305 TI - [The peculiarities of medical support of Home Affairs National Committee Forces of USSR in protection of front-line Red Army rear during the years of Great Patriotic War]. PMID- 16259306 TI - [The patriarch of our medicine (devoted to the 85th anniversary of F. I. Komarov's birth)(Interview by L L Galin and F I Zlain )]. PMID- 16259307 TI - [The specialists' certification as the factor of development of Russian Federation Armed Forces medical service]. PMID- 16259308 TI - Laser acupuncture for mild to moderate depression in a primary care setting--a randomised controlled trial. AB - OBJECTIVE: Depression is a major public health problem. There is some evidence supporting the effectiveness of needle acupuncture in its treatment. Laser stimulation, regarded here as a modality of acupuncture, is non-invasive and therefore free of infection risk; and it is acceptable to patients with needle phobia. The technique is relatively easily learned by community-based general practitioners. It is also amenable to sham control and therefore double-blinding in clinical trials. A double-blind randomised controlled trial was conducted to test the efficacy of low level laser acupuncture in mild to moderate depression. METHODS: Thirty patients with depression were randomised to receive either active or inactive laser treatment. The laser unit could be switched to one of two settings. One switch position delivered active laser acupuncture and the other was inactive (sham). In the active mode, 0.5J was delivered to each of six to eight individually tailored acupuncture sites per visit. All patients were treated twice weekly for four weeks then weekly for a further four weeks. The patients and the acupuncturist were both blinded to conditions. Outcome was assessed using the Beck Depression Inventory at baseline, weeks four and eight during treatment, and at 4 and 12 weeks following the treatment. RESULTS: At the end of the treatment period, Beck Depression Inventory scores fell from baseline by 16.1 points in the intervention group and by 6.8 points in the sham control group (P<0.001). The difference showed only a trend four weeks later, but was again significant after 12 weeks (P=0.007). Laser acupuncture was well tolerated with transient fatigue as the most common adverse effect. CONCLUSION: Laser acupuncture may be worth further investigation as a treatment for mild to moderate depression in primary care. PMID- 16259309 TI - Short term reactions to acupuncture--a cross-sectional survey of patient reports. AB - OBJECTIVE: To explore the type and frequency of short term reactions associated with a single acupuncture treatment. METHODS: As part of recruitment to a large scale prospective survey of the safety of acupuncture, 9408 consecutive patients each completed one survey form soon after receiving treatment with acupuncture, and returned it directly to the research centre. On this form, patients were asked to report on a range of possible short term reactions relating to their most recent acupuncture treatment using a checklist of options. RESULTS: At least one short term reaction to acupuncture during or immediately after treatment was reported by 94.6% (CI 94.2 to 95.1) of patients, an average of 1.8 reactions per patient. The most common experiences reported were feeling 'relaxed' (79.1%) followed by feeling 'energised' (32.7%). A total of 24.4% of patients reported 'tiredness' or 'drowsiness', with obvious implications for safety if the patient intended to drive after treatment. 'Negative' reactions, such as pain and bruising at the site of needling, were reported by 29.7% of patients who were more likely to be female patients (OR 1.58), patients under the age of 40 (OR 1.62), patients who had consulted their GP or hospital specialist beforehand (OR 1.30), patients consulting their acupuncturist for the first time (OR 1.24), and patients treated by an acupuncturist with less than two years' experience since qualification (OR 1.24). Only 13 patients were unwilling to have acupuncture again as a result of these short term reactions. CONCLUSION: In this large cross sectional study, extensive patient reports showed that 'positive' reactions to acupuncture treatment were very common. Tiredness, drowsiness and a range of 'negative' reactions were also frequently reported. Almost all patients were willing to experience these reactions again. PMID- 16259310 TI - A review of myofascial pain and fibromyalgia--factors that promote their persistence. AB - Chronic muscle pain (myalgia) is a common problem throughout the world. Seemingly simple, it is actually a difficult problem for the clinician interested in determining the aetiology of the pain, as well as in managing the pain. The two common muscle pain conditions are fibromyalgia and myofascial pain syndrome. Fibromyalgia is a chronic, widespread muscle tenderness syndrome, associated with central sensitisation. It is often accompanied by chronic sleep disturbance and fatigue, visceral pain syndromes like irritable bowel syndrome and interstitial cystitis. Myofascial pain syndrome is an overuse or muscle stress syndrome characterised by the presence of trigger points in muscle. The problem these syndromes pose lies not in making the diagnosis of muscle pain. Rather, it is the need to identify the underlying cause(s) of persistent or chronic muscle pain in order to develop a specific treatment plan. Chronic myalgia may not improve until the underlying precipitating or perpetuating factor(s) are themselves managed. Precipitating or perpetuating causes of chronic myalgia include structural or mechanical causes like scoliosis, localised joint hypomobility, or generalised or local joint laxity; and metabolic factors like depleted tissue iron stores, hypothyroidism or Vitamin D deficiency. Sometimes, correction of an underlying cause of myalgia is all that is needed to resolve the condition. PMID- 16259311 TI - A model for acupuncture training in primary care. AB - A recent innovation in acupuncture training at a clinic in a primary care practice is reported. The trainer is a General Practitioner within the practice and the trainee an academic nurse. The semi-structured training programme is outlined and benefits and challenges to both trainer and trainee are discussed. The patients generally welcomed the scheme. We consider that the programme could provide a model for similar-training initiatives in the future. PMID- 16259312 TI - Introducing voluntary donations to fund primary care acupuncture--a user survey. AB - An acupuncture clinic has been running in general practice without any funding for nine years. However, reorganisation of the health service meant that it had to become self-funding. A scheme for voluntary donations was introduced, with a target of l15 per treatment. In the first six months, the scheme covered the clinic's costs. A user survey was undertaken to elicit patients' views on the scheme. The majority of patients thought that the scheme was a good idea, and the amount about right. However, a few patients found payment difficult, and stated that they would be less likely to use the service for that reason, or feel guilty about using it without paying. This information should be used to try to obtain public funding through public funds in the future. PMID- 16259314 TI - Acupuncture-induced galactorrhoea--a case report. AB - A case of acupuncture-induced galactorrhoea in a healthy subject is described. The acupuncture was performed at a non-traditional site using a periosteal technique and galactorrhoea occurred on two separate occasions. Galactorrhoea following acupuncture has been reported on one previous occasion. PMID- 16259313 TI - Acupuncture in the treatment of post viral fatigue syndrome--a case report. AB - This case report concerns the treatment of post viral fatigue (chronic fatigue syndrome) with electroacupuncture. This condition is particularly difficult to treat whether using conventional or complementary therapy. Whilst the treatment did not cure the patient, it appears to have facilitated her return to work and markedly improved her symptoms. There are few publications on acupuncture treatment of this condition and the approach used here has not been reported previously. PMID- 16259315 TI - Pharmacological potential of endothelin receptors agonists and antagonists. AB - Endothelins are potent predominantly vasoconstricting agents that act as local autocrine and paracrine mediators. Endothelin-1 is the most potent and sustained vasoconstrictor and pressor substance yet identified. Abnormalities of the endothelin system occur in a range of diseases associated with vasoconstriction, vasospasm, and vascular hypertrophy. ET receptor antagonists were until recently regarded as drugs of great promise in patients with congestive heart failure, pulmonary hypertension and others. The aim of this article is a survey of compounds that affect the endothelin receptors and clinical trials with these agents. PMID- 16259316 TI - The diagnostic role of urinary N-acetyl-beta-D-glucosaminidase (NAG) activity in the detection of renal tubular impairment. AB - The kidney function can be assessed by a number of methods. The urinary excretion of enzymes, in particular N-acetyl-beta-D-glucosaminidase (NAG), is considered a relatively simple, cheap, fast and non-invasive method in the detection and follow-up of renal tubular function under various conditions. The determination of urinary NAG provides a very sensitive and reliable indicator of renal damage, such as injury or dysfunction due to diabetes mellitus, nephrotic syndrome, inflammation, vesicoureteral reflux, urinary tract infection, hypercalciuria, urolithiasis, nephrocalcinosis, perinatal asphyxia, hypoxia, hypertension, heavy metals poisoning, treatment with aminoglycosides, valproate, or other nephrotoxic drugs. This paper gives an overview of the current use of urinary NAG in the detection of renal injury. PMID- 16259318 TI - The influence of the time of antidotal treatment administration on the potency of newly developed oximes to counteract acute toxic effects of tabun in mice. AB - (1) The influence of the time of administration of antidotal treatment consisting of anticholinergic drug (atropine) and newly developed oxime (K027 or K048) on its effectiveness to eliminate tabun-induced lethal toxic effects was studied in mice. (2) The therapeutic efficacy of antidotal treatment of tabun-induced acute poisoning depends on the time of its administration regardless of the choice of the oxime. (3) Our results show that both oximes studied (K027, K048) are able to sufficiently eliminate lethal effects of tabun. Nevertheless, their efficacy significantly decreases when they were administered 5 min after tabun poisoning. (4) The findings support the hypothesis that both newly developed oximes appear to be suitable oximes to counteract acute toxicity of tabun although their ability to eliminate lethal toxic effects of tabun significantly decreases with prolonged time interval between tabun challenge and antidotal treatment administration. PMID- 16259317 TI - A comparison of the potency of the oxime HLo-7 and currently used oximes (HI-6, pralidoxime, obidoxime) to reactivate nerve agent-inhibited rat brain acetylcholinesterase by in vitro methods. AB - (1) The efficacy of the oxime HLo7 and currently used oximes (pralidoxime, obidoxime, HI-6) to reactivate acetylcholinesterase inhibited by various nerve agents (sarin, tabun, cyclosarin, VX) was tested by in vitro methods. (2) Both H oximes (HLo-7, HI-6) were found to be more efficacious reactivators of sarin and VX-inhibited acetylcholinesterase than pralidoxime and obidoxime. On the other hand, their potency to reactivate tabun-inhibited acetylcholinesterase is very low and does not reach the reactivating efficacy of obidoxime. In the case of cyclosarin, the oxime HI-6 was only found to be able to sufficiently reactivate cyclosarin-inhibited acetylcholinesterase in vitro. (3) Thus, the oxime HLo-7 does not seem to be more efficacious reactivator of nerve agent-inhibited acetylcholinesterase than HI-6 according to in vitro evaluation of their reactivation potency and, therefore, it is not more suitable to be introduced for antidotal treatment of nerve agent-exposed people than HI-6. PMID- 16259319 TI - Bone marrow necrosis: a rare complication of herbal treatment with Hypericum perforatum (St. John's wort). AB - A 22-year-old man presented with fever and ulceration of the oral mucosa. The patient had pharyngeal and esophageal candidiasis. For the 3 weeks prior to presentation, he had been taking approximately 1000 mg/day of flowering herbs (Hypericum perforatum L, known as St. John's wort) for treatment of depression. A complete blood count on the first day of hospitalization showed agranulocytosis and normocytic anemia. His condition worsened, and he developed progressive dysphagia. A bone marrow biopsy on day 3 revealed bone marrow necrosis. After the diagnosis was established (day 3 of hospitalization), treatment with granulocyte colony-stimulating factor 48 U/day, intravenous immunoglobulin 400 mg/kg, and amphotericin B 100 mg/day was initiated. The patient did not respond, and died within one week of the diagnosis. This cases suggests that Hypericum species may lead to severe hematologic toxicity, with conditions involving bone marrow necrosis. PMID- 16259320 TI - Use of two parallel oxygenators in a very large patient (2.76 m2) for an acute "A" dissecting aortic aneurysm repair. AB - The very large patient (weight 142 kg, height 197 cm, body surface 2.76 m2) was referred to acute operation with dissecting type A ascending aortic aneurysm. The calculated blood flow was 6.63 l/min. To anticipate potential difficulties with perfusion and oxygenation two oxygenators connected in parallel were incorporated into the circuit. Bentall procedure with ACB to the RCA was performed. The perfusion was uneventful. Bypass time was 259 minutes, cross clamp time 141 minutes, circulatory arrest 7 minutes. The highest oxygenators gas flow was 2.6 l/min with maximum FiO2 0.42. The use of two in parallel connected oxygenators is a very effective, easy and safe method in such extreme perfusions, offering to the perfusionist a great reserve of oxygenator output. PMID- 16259321 TI - Mental retardation in childhood: clinical and diagnostic profile in 100 children. AB - Mental retardation (MR) is present in 2-3% of individuals in the general population, either as an isolated finding, or as part of an underlying disorder. It is the reason for a substantial part of referrals of patients and families to the pediatric genetic counseling units. MR may be genetically determined, or due to environmental (including perinatal) influences. Despite the thorough examination, physicians often are unable to define the etiology, in at least 30 50% of cases. This report presents the experience with 100 consecutive children with mental retardation, admitted to the Section of clinical genetics of the University pediatric hospital in Sofia over one year period. According to the routinely used classification, the patients' disease states have been subgrouped into genetic (35), multifactorial (3), environmental (3), and of unknown etiology (59). The research protocol included careful dysmorphologic, neurologic and developmental assessment, as well as cytogenetic, biochemical and molecular genetic testing. The rate of the diagnostic yield of the whole group was 41%. The discussion includes clinical examples and literature reports. PMID- 16259322 TI - A follow-up study on 12 prenatally diagnosed boys with Klinefelter syndrome. AB - Longitudinal follow-up data on males with Klinefelter syndrome are still scarce. In the present study we collected data on the general and psychosocial development of 12 prenatally diagnosed boys with Klinefelter syndrome. PMID- 16259323 TI - M-FISH applications in clinical genetics. AB - Until recently, presence of de novo marker or derivative chromosomes was quite problematic for genetic counseling especially in prenatal diagnosis, because characterization of marker and derivative chromosomes by conventional cytogenetic techniques was nearly impossible. However, recently developed molecular cytogenetic technique named Multicolor Fluorescence in Situ Hybridization (M FISH) which paints all human chromosomes in 24 different colors allows us to characterize marker and derivative chromosomes in a single hybridization. In this study, we applied M-FISH to determine the origin of 3 marker and 3 derivative chromosomes. Marker chromosomes were found to originate from chromosome 15 in two postnatal and one prenatal case. Of these, one of the postnatal cases displayed clinical findings of inv dup (115) syndrome and the other of infertility, and the prenatal case went through amniocentesis due to the triple test results. Karyotypes of the patients with derivative chromosomes were designated as 46,XY,der (21)t(1;21)(q32;p11), 46,XX,der(8)t(8;9)(p23;p22) and 46,XX,der(18)t(18;20)(q32;p11.2) according to cytogenetic and M-FISH studies. All of the M-FISH results were confirmed with locus specific or whole chromosome painting probes. The case with der (8)t(8;9) had trisomy 9(p22-pter) and monosomy 8(p23-pter) due to this derivative chromosome. The case with der(18)t(18;20) had trisomy 20(p11.2-pter) and monosomy 18(q32-qter). Parental origins of the derivative chromosomes were analyzed using microsatellite markers located in the trisomic chromosomal segments. Patients' clinical findings were compared with the literature. PMID- 16259324 TI - Proximal 6q interstitial deletion without severe mental retardation. AB - Deletions of the proximal portion of the long arm of chromosome 6 are rare, and the patients reported in the literature have been described as having significant mental retardation, often in the severe to profound range. We report on a young girl with a proximal 6q interstitial deletion [46, XX, del (6) (q1 3q15)] who exhibits the typical morphological and neurological manifestations except that the degree of language and non-language cognitive delay is mild. Cognitive development has been assessed using the Capute Scales. Gross motor development has been assessed clinically using standard milestones. At 34 months of age, she was found to have severe gross motor delay, with mildly delayed non-language cognitive abilities and expressive language abilities. Her receptive language skills tested in the average range. In recent years. several forms of autosomal dominant drusen and macular degeneration have been mapped to 6q14, and substantial variability in clinical expression has been described. Serial ophthalmologic examinations have not detected any drusen or atrophic macular changes in our patient. To our knowledge, this is the first description of a child with proximal 6q interstitial deletion in the documented absence of severe cognitive deficiency. This information may be important to clinicians who counsel families of infants determined to have proximal 6q interstitial deletions. Because the deleted region contains the area to which autosomal dominant drusen and macular degeneration has been mapped, we believe that this child is at risk for visual loss, and she will continue to be monitored closely. PMID- 16259325 TI - Report of a patient with a trisomy of chromosome region 20q11.2-->20q12 and characterization with FISH. AB - We report on a 16-month-old boy presenting with psychomotor retardation, craniofacial anomalies and severe vision deficit. Analysis of GTG-banded chromosomes showed that the patient had extra chromosomal material in the long arm of one chromosome 20. This chromosome aberration was further characterized with FISH using a chromosome 20 specific paint and band-specific probes. A partial trisomy 20q was shown to be present, the karyotype being 46, XY, dup (20) (q11.2q12). The cytogenetic and clinical findings are compared with cases previously reported in the literature. PMID- 16259326 TI - Partial trisomy 11q syndrome (11q23.1-->11qter) due to de novo t (11q; 13q) detected by multicolor fluorescence in situ hybridisation. AB - In this report we describe the identification of a de novo 46, XX, 13q + by multicolour fluorescence in situ hybridisation (M-FISH), as a partial distal 11q trisomy (11q23.1-->11qter). The clinical phenotype association with this distal 11q trisomy is briefly reviewed. PMID- 16259327 TI - Robinow Syndrome: a case report. AB - We report a case with Robinow syndrome which has been rarely reported in the literature. A male newborn who had fetal face appearance (broad and prominent forehead, hypertelorism, small saddle nose, anteverted nostrils, glabellar nevus flammeus, malar hypoplasia, down-turned mouth and retrognathia), mesomelic limb shortening, hemivertebra and genital hypoplasia was diagnosed as Robinow syndrome. Elevated levels of both basal and stimulated testosterone and dihydrotestosterone were found along with normal baseline levels of gonadotropins. These endocrinologic studies were suggestive for an androgen insensitivity. Mental and motor development of the infant were normal at 3 and 6 months of age. Because of the high level of consanguineous marriages in Turkey, we may expect a higher incidence of the autosomal recessive form of the syndrome. This gives a high recurrence risk and makes prenatal diagnosis an important option for future pregnancies in the families. PMID- 16259328 TI - Fetal therapy and cytogenetic testing: prenatal detection of chromosome aberration during thoracocentesis for congenital chylothorax by karyotyping from pleural effusion fluid and review of the literature. AB - This report serves to emphasize the necessity of rapid cytogenetic testing during fetal therapy for congenital hydrothorax and to review the literature. A 31-year old primigravid woman was noted to have bilateral fetal hydrothorax, polyhydramnios, and preterm labor at 32 weeks' gestation. Echo-guided thoracocentesis was performed to draw 50 ml of golden/yellow pleural effusion fluid and 500 ml of amniotic fluid. Cytogenetic analysis of the lymphocytes obtained from the pleural effusion fluid revealed a karyotype of 47, XY, + 21. The pleural effusion fluid was predominantly lymphocytic and positive for the Rivalta test. A sonographic examination at 33 weeks' gestation revealed recurrent pleural effusion, but the woman refused repeat thoracocentesis and tocolytic management. A 2,568-g male baby with characteristic phenotypic findings of Down syndrome was delivered vaginally and expired after birth. The present case reinforces the notions that fetuses with congenital chylothorax are at risk for chromosomal abnormalities, and drainage of pleural effusion must include a rapid diagnosis of fetal karyotype. The cytogenetic information acquired is useful for genetic counseling and perinatal obstetric management. PMID- 16259329 TI - The phenotypic effect of Hb G-Coushatta [beta22 (B4) Glu-Ala] and association with IVS.II.1(G-A) in a Turkish family. PMID- 16259330 TI - Alterations in the GJB3 and CLDN14 genes in families with nonsyndromic sensorineural hearing loss. PMID- 16259331 TI - Maternal diabetes and neural tube defects: prenatal diagnosis of lumbosacral myelomeningocele, ventriculomegaly, Arnold-Chiari malformation and foot deformities in a pregnancy with poor maternal metabolic control, and review of the literature. PMID- 16259332 TI - A rare chromosomal rearrangement [inv (2)(p11.q13)] in a 37-year-old normal male and 45,X/46,X,+ mar in his daughter. PMID- 16259334 TI - [Modern treatment of rheumatoid arthritis]. AB - Modern treatment of rheumatoid arthritis. Rheumatoid arthritis is a chronic inflammatory disease of unknown etiology, which is characterised by pain, loss of capability to work, in severe cases the life expectancy is also reduced. The disease cannot be cured with current therapeutic possibilities, but complaints can be reduced, the destruction can be retarded. The treatment is more efficient in the early stage of the disease, but early diagnosis is difficult because of insidious onset and limited sensitivity of diagnostic methods. The complaints can be alleviated by nonsteroidal anti-inflammatory drugs and transient glucocorticoid treatment, but risk of continuous glucocorticoid therapy is significant. To prevent structural damage disease modifying antirheumatic drugs are used. Out of these methotrexate is the most effective and it is well tolerated. Destruction of the joints is the consequence of inflammation, so intensity of drug treatment must be adjusted to inflammatory activity. For monitoring in clinical practice the composite index disease activity score is recommended. To achieve the reduction of inflammatory activity the dosage of disease modifying drugs can be increased, they can be switched or combined, and continuous glucocorticoid treatment can be started. In cases refractory to conventional treatment it is possible to inhibit the activity of proinflammatory cytokines, which play a pivotal role in pathomechanism of rheumatoid arthritis. In synovitis limited to one joint intraarticular glucocorticoid injection can be given, in refractory cases synovectomy is indicated. Destruction of the joints can be partially corrected by exercise, orthoses and after all with surgery. PMID- 16259333 TI - [Kaposi's sarcoma]. AB - In recent years large amounts of findings have accumulated about Kaposi's sarcoma, a virus induced angioproliferative disorder appearing in four clinical forms: classical, epidemic, endemic and iatrogenic, as it has been in focus of not only from the dermatologic but also from the viral tumorgenesis perspective. The common characteristics are the histopathological appearance, the causative role of the human herpesvirus 8 and the similar clinical picture (bluish-red macules, papules, and nodes). Frequency of the distinguished clinical forms differs with geographical location. Viruses, genetic -, and environmental factors have been shown to play a role in the pathomechanism of the disease, of which the most important is the human herpesvirus-8. The mechanisms by which viral proteins and virus infection enhance tumorgenesis and alter immune functions directed at cells have been studied in detail. During the initiation of tumorgenesis, virus induced viral and host cell products (cytokines, receptors and oncogens) initiate inflammatory and angiogenic polyclonal cell proliferation, which later, by the synergistic action of other viruses and/or environmental factors, give rise to malignant proliferation and allow the selected cell to clonally expand and behave like a true malignant tumor. In light of newly published results the authors not only present the clinical appearances and summarize diagnostic possibilities and the pathomechanism of the disease, but also give a thorough overview of the therapeutic tools of Kaposi's sarcoma, and share their experiences obtained during the follow-up of classical Kaposi's sarcoma patients. PMID- 16259335 TI - [Cardiovascular consequences of thyroid diseases, their prevalence and treatment, particularly in cases with subclinical hyper- and hypothyroidism]. AB - Cardiovascular consequences of thyroid diseases, their prevalence and treatment, particularly in cases with subclinical hyper- and hypothyroidism. The aim of the study was to draw attention to an association between the thyroid and cardiovascular diseases. The main topic was to lay emphasis on the importance of cardiovascular diseases caused by subclinical hyper- and hypothyroidism in the relation to the practice. The subclinical states precede the overt hyper- and hypothyroidism, and they are often present during their treatments. The changes in the levels of thyrotropin demonstrating subclinical thyroid diseases, could be detected in different non-thyroid diseases and resulted from the effect of some drugs. Particularly, the subclinical thyroid diseases become more frequent in older age. In the background of the high prevalences of atrial fibrillation, hypertension and psychosomatic events subclinical hyperthyroidism could be revealed. Subclinical hypothyroidism is characterized by an increased prevalence of elevated serum lipid levels, atherosclerosis, ischemic heart disease and hypertension often associating with the presence of anti-thyroid antibodies. It is important to reveal subclinical thyroid diseases in time for the effective treatment and for stopping of the cardiovascular damages before manifestations of cardiovascular diseases. The paper gives advice for the practice and the rational management. At the end, a survey of the association between thyroid and heart diseases in own department of internal medicine at the last three years is given. PMID- 16259336 TI - [Hepatorenal syndrome]. AB - Hepatorenal syndrome. Hepatorenal syndrome is a functional renal failure in state of portal hypertension caused by splanchnic vasodilatation that brings decrease of renal perfusion. The inductive factors include acute alcoholic hepatitis, non adequate diuretic therapy, infections and bleeding from upper parts of gastrointestinal tract. Recognition of pathophysiological mechanisms of hepatorenal syndrome helped the elaboration of therapeutic schemes. The treatment is successful in improvement of renal functions in many patients but it is very expensive. PMID- 16259337 TI - [Five hundred lines on the margin of the "Bela Johan case"]. PMID- 16259338 TI - Pelvic fractures in a level I trauma center: a test case for the efficacy of the evolving trauma system in Israel. AB - BACKGROUND: Pelvic fracture is a severe and life-threatening injury that requires treatment by a dedicated team. One of the goals of a nationwide trauma system is to provide appropriate medical care for such injuries. OBJECTIVES: To use pelvic fractures as a test case for the efficiency of the Israeli trauma system, as reflected in the experience of our medical center. METHODS: Data were obtained from the medical charts of all cases of pelvic fractures admitted to our medical center between 1987 and 1999. We obtained demographic data, information on the cause of injury, fracture classification, co-injuries and Injury Severity Score, treatment strategies, and mortality rate. RESULTS: Altogether, 808 patients with pelvic injuries were treated in our medical center. The most common cause of injury was motor vehicle accidents (51%). Pelvic fractures without acetabular involvement were diagnosed in 58% of patients and isolated acetabular fractures in 32%, while 10% sustained combined injuries to the pelvic ring and the acetabulum. The overall rate of operative stabilization was 34%. The majority of patients had associated injuries, mostly additional musculoskeletal injuries. Altogether, 13% were referred from Level II/III trauma centers. We observed an increase in the total number of local admissions, in the percentage of referred patients and in the percentage of operated patients during the study period. The observed mortality rate was 5%. CONCLUSIONS: Our results show a more than twofold increase in the percentage of referred patients following the designation of a Level I trauma center. These referrals are due not only to the designation as a Level I trauma center, but also to the presence of a dedicated team of pelvic fracture specialists, available 24 hours a day. In addition, a larger percentage of patients undergo surgery for internal fixation of pelvic fractures, in accordance with current worldwide trends. PMID- 16259339 TI - The role of pelvic fractures in the course of treatment and outcome of trauma patients. AB - BACKGROUND: Pelvic fracture poses a complex challenge to the trauma surgeon. It is associated with head, thoracic and abdominal injuries. As pelvic fracture severity increases so does the number of associated injuries and the mortality rate. OBJECTIVES: To report our experience in the treatment of pelvic fractures. METHODS: Between October 1998 and September 2001, 78 patients with pelvic fractures were admitted to our hospital. The age range of the 56 male and 22 female patients was 16-92 (mean 42 years). The cause of injury was road accident in 52 patients, fall from a height in 15, a simple fall in 9, and gunshot wounds in 2 patients. The Glascow Coma Scale score on arrival at the hospital was 3-15 (average 12). Twenty-five patients (32%) were admitted to the intensive care unit, 38 (48%) to the orthopedic department, 5 (6.4%) to neurosurgery and the remainder to a surgical department. RESULTS: Twenty-six patients (33.3%) received blood transfusion in the first 24 hours. Of the 25 patients with associated head trauma, 6 had intracranial bleeding (32%); 29 patients (37%) had associated chest trauma, 28 (35.9%) had associated abdominal trauma, 16 (20.5%) had vertebral fractures and 40 (51.2%) had associated limb fractures. Pelvic angiography was performed in 5 patients (6.4%), and computed tomography-angiography of the cervical arteries and chest was performed in 1 and 5 patients respectively. Overall, a CT scan was performed in 56 patients (71.8%), of whom 25 (32%) had a pelvic CT on admission. Injury Severity Score was 4-66 (median 20). Laparotomy was performed in 14 patients (18%), spinal fusion in 5 (6.4%), limb surgery in 16 (20.5%), cranial surgery in 4 (5.02%), pelvic surgery in 10 (12.8%), chest surgery in 3 (3.85%), and facial surgery in 2 patients (2.56%). Seven patients (9%) died during the course of treatment. CONCLUSIONS: Pelvic fracture carries a high morbidity rate. Associated chest, abdomen and limb injuries are often encountered. A multidisciplinary approach is needed to improve survival and outcome in patients with pelvic fractures. PMID- 16259340 TI - Psychological correlates of idiopathic intracranial hypertension. AB - BACKGROUND: Idiopathic intracranial hypertension, also known as pseudotumor cerebri, is a relatively rare disorder of increased intracranial pressure >250 mm water, with a normal neuroimaging and normal cerebrospinal fluid content. OBJECTIVES: To examine whether hostility, anxiety and autobiographical memory (a correlate of depression) are associated with IIH. METHODS: Using a case-control cross sectional design, 20 patients with IIH were compared with 9 healthy controls of similar age, weight and height, and 11 headache controls. Patients were assessed for hostility and anxiety. The Autobiographical Memory Test was used to assess episodic memories. RESULTS: The IIH group reported significantly more anxiety and more general episodic memories than the healthy controls, but not the headache control group. The headache control patients reported more general memories than did the healthy controls. CONCLUSIONS: Patients with headaches, whether of general origin or related to IIH, have a poor psychosocial profile. While the study design does not permit any conclusions regarding causality, our results support the need to consider psychological factors in evaluating and treating IIH and headache patients. PMID- 16259341 TI - Measurement of electrical skin impedance of dermal-visceral zones as a diagnostic tool for inner organ pathologies: a blinded preliminary evaluation of a new technique. AB - BACKGROUND: As complementary and alternative medicine is gaining popularity among health consumers, diagnostic screening tools based on neuroreflexology are also being developed. These techniques, which are based on the rationale that measurement of electrical impedance of specific dermatomes reflects corresponding internal organ pathologies, have not yet been the subject of conventional scientific research. OBJECTIVES: To determine the effectiveness of a neuroreflexology-based screening test, specifically the Medex device (Medex Screen Ltd.), for diagnosing patients undergoing conventional internal organ assessment, in a hospital setting. METHODS: Patients admitted to an internal medicine department, who met the inclusion criteria and agreed to participate, underwent conventional medical evaluation that included past medical history and physical examination. Another examination was conducted by a second physician using the Medex device to determine internal organ pathologies. A third researcher compared the actual "conventional" diagnosis with the Medex device output using standard statistical analysis. RESULTS: Overall, 150 patients participated in the study. Correlation was significant for all categories (P < 0.01) except for blood and lymphatic disease. A high sensitivity (>70%) was measured for cardiovascular, respiratory, gastrointestinal and genitourinary diseases. The highest measure of agreement, as represented by the Cohen-Kappa factor, was found for respiratory disease (0.57). CONCLUSIONS: Although the exact mechanism is not entirely clear, measurement of electroskin impedance of dermal visceral zones has the potential to serve as a screening tool for inner organ pathologies. Further research should be conducted to create more evidence to support or dispute the use of this technique as a reliable diagnostic tool. PMID- 16259342 TI - Characteristics and outcomes of ninth and tenth decade patients hospitalized in a sub-acute geriatric hospital. AB - BACKGROUND: Population structures are changing across the western world, with particularly rapid growth in the number of very old people. Life expectancy has been increasing gradually over years, resulting in a larger subpopulation of people aged 90 and over. OBJECTIVES: To describe the sociodemographic, medical and functional characteristics of people aged 80-90 and 90+ who were admitted to a sub-acute geriatric hospital and to compare the hospitalization outcomes between these subgroups. METHODS: We compared the demographic and clinical data (extracted by means of chart review) of two groups of elderly who were admitted to the Reuth Medical Center during 2001-2002: those aged 90+ and those 80-89. Among survivors, the main outcome measures at discharge were mortality rate, functional ability, and place of residence. RESULTS: The study included 108 patients who were admitted to different divisions of Reuth: 55 patients aged 90+ and 53 aged 80-90. The mortality rate was significantly elevated in the older age group (49.1% vs. 28.1% in the younger age group). On multivariate analysis, the most important prognostic factors for mortality were incontinence (odds ratio 3.45) and being dependent before admission (OR 4.76). Among survivors, an association was found between being incontinent and dependent before hospitalization, and being dependent on discharge. CONCLUSIONS: The main prognostic factors for mortality and functional outcome in patients admitted to a non-acute geriatric hospital are incontinence and functional state prior to admission, and not age per se. PMID- 16259343 TI - Pediatric advanced life support (PALS) courses in Israel: ten years of experience. AB - BACKGROUND: The Pediatric Advanced Life Support course of the American Heart Association/American Academy of Pediatrics was established in Israel in 1994 and has since been presented to over 3,108 medical and paramedical personnel. OBJECTIVES: To assess the achievements of participants in the PALS course, as a cohort and by professional group, and their evaluations of different aspects of the course; and to describe the educational modifications introduced to the course since its introduction in Israel based on our teaching experience. METHODS: The study sample consisted of physicians, nurses and paramedics from all areas of Israel who registered for PALS between January 2001 and December 2003. Participants took a standardized test before and after the course; a score of 80 or higher was considered a pass. On completion of the course, participants were requested to complete a 24-item questionnaire evaluating the quality of the course as a whole, as well as the lectures, skill stations, and instructors' performance. Items were rated on a 5-point scale. Results were analyzed using the BMPD statistical package. RESULTS: Altogether, 739 subjects participated in 28 courses: 13 attending (in-hospital) physicians (1.8%), 89 community pediatricians (12%), 124 residents (16.8%), 304 nurses (41.1%), and 209 paramedics (28.3%). About half (48.9%) were hospital-based, and about half (47.9%) had no experience in emergency medicine. A passing grade was achieved by 89.4% of the participants; the mean grade for the whole sample was 87.2%. The mean test score of the residents was significantly better than that of the nurses (P < 0.05) and pediatricians (P < 0.01). The median evaluation score for four of the five stations was 5, and the mean overall score for all items was 4.56 (range by item 3.93-4.78). CONCLUSIONS: PALS was successfully delivered to a large number of healthcare providers in various professional groups with very good overall achievements and high participant satisfaction. It significantly increased participants' knowledge of pediatric resuscitation. We therefore recommend the PALS course as an educational tool in Israel. PMID- 16259344 TI - Quality of diabetes care in the community: a cross-sectional study in central Israel. AB - BACKGROUND: Good care of the diabetic patient reduces the incidence of long-term complications. Treatment should be interdisciplinary; in the last decade a debate has raged over how to optimize treatment and how to use the various services efficiently. OBJECTIVES: To evaluate the quality of care of diabetic patients in primary care and diabetes clinics in the community in central Israel. METHODS: We conducted a retrospective cross-sectional study of a random sample of 209 diabetic patients in a district of the largest health management organization in Israel. Patients were divided into two groups - those treated only by their family physician and those who had attended diabetes clinics. Data included social demographics, medications, risk factors, quality of follow-up, laboratory tests, quality of diabetes and blood pressure control, and complications of diabetes. RESULTS: Of the 209 patients 38% were followed by a diabetes clinic and 62% by a family physician. Patients attending the specialist clinic tended to be younger (P= 0.01) and more educated (P= 0.017). The duration of their diabetes was longer (P < 0.01) and they had more diabetic microvascular complications (P= 0.001). The percentage of patients treated with insulin was higher among the diabetes clinic patients (75% vs. 14%, P= 0.0001). More patients with nephropathy received angiotensin-converting enzyme inhibitors in the diabetes clinic (94% vs. 68%, P= 0.02). Follow-up in the specialist clinic as compared to by the family physician was better in the areas of foot examination (P< 0.01), fundus examination (P= 0.0001), and hemoglobin A1c testing (P= 0.01). On a regression model only fundus examination, foot examination and documentation of smoking status were significantly better in the diabetes clinic (P< 0.05). CONCLUSION: There is still a large gap between clinical guidelines and clinical practice. Joint treatment of diabetic patients between the family physician and the diabetes specialist may be a proposed model to improve follow-up and diabetes control. This model of treatment should be checked in a prospective study. PMID- 16259345 TI - Advances in the treatment of pelvic fractures. PMID- 16259346 TI - Screening, the everlasting quest for the needle in the haystack. PMID- 16259347 TI - Non-conventional diagnostic tools: do we really need them? PMID- 16259348 TI - The advanced XXXX life support ("AXLS") contribution to emergency medicine in Israel. PMID- 16259349 TI - Allopurinol-induced DRESS syndrome. AB - Hyperuricemia is present in approximately 5% of the population, the vast majority of whom are asymptomatic and at no clinical risk. Complications, including renal calculi, uric acid nephropathy and gout, occur in a small proportion of patients. Allopurinol, an analog of hypoxanthine, has been widely used in clinical practice for over 30 years for the treatment of hyperuricemia and gout. Two percent of patients taking this medication develop a mild exanthema. A syndrome characterized by exfoliative dermatitis, hepatitis, interstitial nephritis and eosinophilia has been described previously. Termed allopurinol hypersensitivity syndrome, its etiology is related to the accumulation of one of the allopurinol metabolites, oxypurinol; clearance of oxypurinol is decreased in the setting of renal insufficiency and the use of thiazide diuretics. The term DRESS syndrome (Drug Rash with Eosinophilia and Systemic Symptoms) was recently introduced to describe a disorder associated with various drugs or viral infections and characterized by similar features. The pathophysiology of allopurinol-induced hypersensitivity, clinical presentation and treatment are reviewed. PMID- 16259350 TI - Moyamoya disease--diagnosis and treatment: indirect cerebral revascularization at the Sheba Medical Center. AB - Moyamoya disease is a cerebral vasculopathy characterized mainly by progressive narrowing of the major intracranial vessels. While more common and having a familial predilection in the Far East, it can also develop in association with some common hereditary diseases and can be acquired after environmental exposure. In the young its manifestations are the result of cerebral ischemia. Adults usually suffer from repeated incidents of intracerebral hemorrhage. Surgical revascularization of ischemic cerebral territories plays a major role in their treatment. We review the literature and present our series of three adult and five pediatric patients; these patients were diagnosed at our institution and treated with indirect revascularization techniques. PMID- 16259351 TI - Rheumatoid arthritis in thalassemia intermedia: coincidence or association? PMID- 16259352 TI - Acute polyarthritis associated with clopidogrel treatment. PMID- 16259353 TI - Improved survival with surgery and systemic chemotherapy for undifferentiated embryonal sarcoma of the liver. PMID- 16259354 TI - Carcinoid tumor and Crohn's ileitis. PMID- 16259355 TI - Vascular compression of the optic chiasm resembling glaucoma-like visual field defects. PMID- 16259356 TI - Coincidence of ankylosing spondylitis, gouty arthritis and chondrocalcinosis articularis. PMID- 16259357 TI - Schistosomal appendicitis. PMID- 16259358 TI - [Immunohistochemical study on expression of iNOS and eNOS during skin incised wound healing in mice]. AB - OBJECTIVE: To investigate the expressions of iNOS and eNOS during skin incised wound healing and the applicability of time-dependent expressions of iNOS and eNOS to timing of wound. METHODS: An incised wound was made in the dorsal skin of mouse. By using S-P method, the expression of iNOS and eNOS were studied in cutaneous incised wound as well as normal skin in control mice. RESULTS: iNOS and eNOS expressed in epidermis, hair follicle, sebaceous gland, and the eNOS were also found in vascular endothelium in control group. Three hours after injuryed, the expression of iNOS and eNOS were detected in polymorphonuclear cells (PMNs) in the wound and peripheral region. From 6h to 24h of wound interval, the expression of NOS isoforms were identified in a large number of infiltrating PMNs and part of mononuclear cells (MNCs), afterwards the MNCs and fibroblastic cells (FBCs) accounted for the most part of the iNOS and eNOS positive cells. In addition, during wound healing, the expressions of iNOS and eNOS were noticed in different level in vascular epithelial cells, especially neonatal capillary vessel in granulation tissue. CONCLUSION: The results suggest that iNOS and eNOS express in polymorphonuclear cells, macrophages and fibroblasts during healing process of skin incised wound in mice. Furthermore, iNOS and eNOS may play pivotal roles in all the events relating to wound healing, and the time-dependent expressions of iNOS and eNOS may possibly be used as new markers for the wound age determination. PMID- 16259359 TI - [Expression of beta-amyloid precursor protein in diffuse axonal injury of rats]. AB - OBJECTIVE: To explore an method for diffuse axonal injury (DAI) diagnosis and injury time estimation, the changes of beta-APP immunoreactivity and to observe the morphology of axonal in different parts of brain after experimental DAI injury. METHODS: The animal models of DAI was established according to the Marmarou's method. Immunohistochemistry and Gless staining were performed to observe the changes of beta-APP expression and the morphology of axon with the time elapsed after the DAI injury. RESULTS: In the brain injury group, the morphologic changes of axon in brain stem were showed as twisted, broken and swellen at 0.5 h, and the myelin sheaths broken could be observed, the retraction ball was found at 12 h. Those morphology changes further progressed at 12h, reached to peack up to 1 d, then repaired at 3 d, and recovered at 10 d; Meanwhile the analysis of beta-APP immunoreactivity was also showed a time dependent difference as fellows: beta-APP expression begin at 3h, increased its immunoreactivity at 12h, reached to maximize at 1 d, decreased after 3 d, returned to basal level at 10 d. CONCLUSIONS: The results suggest that beta-APP immunohistochemistry combine with Gless staining be sensitive methods for DAI diagnosis, they could discover the time-dependent changes of the axonal morphology.The changes beta-APP are quite regular and could be used for timing DAI injury. PMID- 16259360 TI - [Lethal anaphylactic shock model induced by human mixed serum in guinea pigs]. AB - OBJECTIVE: To establish an anaphylactic shock model induced by human mixed serum in guinea pigs. METHODS: Eighteen guinea pigs were divided into two groups: sensitized and control, The sensitized group were immunized intracutaneously with human mixed serum and then induced by endocardiac injection after 3 weeks. RESULTS: Symptoms of anaphylactic shock appeared in the sensitized group. The level of serum IgE were increased in the sensitized group significantly. CONCLUSIONS: An animal model of anaphylactic shock wer established successfully. It provide a tool for both forensic study and anaphylactic shock therapy. PMID- 16259361 TI - [Expression of c-Fos in rats organs after electrical injury]. AB - OBJECTIVE: To study the expression of c-fos in organs after rats electrified. METHODS: The brain, lung, heart, liver, spleen, kidney, muscle of electrified limb, and cutis of electrified limb of all experimental rats and those organs of control groups were dissected to detect the expression of c-fos by using immunohistochemistry staining, and the staining intensity were assessed by image analysis system. RESULTS: The expression of c-fos was observed in brain, heart, liver, lung, kidney and muscle in electrified directly group, the amount of expression reached peak at 2 h after electrified and decreasing at 8 h after electrified, and the expression showed faintness in electrified at the immediate after death group. The expressions of c-fos in spleen and cutis is negative in all groups. The expression of c-fos in all detected organs was negative in other rats that were electrified after death. CONCLUSION: c-fos can be regard as a target in identifying electrical injury, whether it formed at antemortem or postmortem. PMID- 16259362 TI - [Correlation between PMI and DNA degradation of costicartilage and dental pulp cells in human being]. AB - OBJECTIVE: To probe the correlation between the postmortem interval (PMI) and the DNA degradation of costicartilage and dental pulp cells in human being after death, and to seek a new method for estimating PMI. METHODS: The image cytometry was used to measure the DNA degradation under different ambient temperatures (30 35 degrees C, 15-20 degrees C) in 0-15 days after death. RESULTS: The average DNA content of two kinds of tissue was degradated with the prolongation of PMI. But there was a plateau period of 0-4 days for dental pulp cells of human being in 15 20 degrees C. There was a high negative correlativity P<0.01 between the average DNA content and PMI. CONCLUSION: PMI could be estimated accurately according to the DNA degradation of costicartilage and dental pulp cells in human being after death. PMID- 16259363 TI - [Research & development on computer expert system for forensic bones estimation]. AB - OBJECTIVE: To build an expert system for forensic bones estimation. METHODS: By using the object oriented method, employing statistical data of forensic anthropology, combining the statistical data frame knowledge representation with productions and also using the fuzzy matching and DS evidence theory method. RESULTS: Software for forensic estimation of sex, age and height with opened knowledge base was designed. CONCLUSION: This system is reliable and effective, and it would be a good assistant of the forensic technician. PMID- 16259364 TI - [Analysis of Doppler ultrasonography in penile artery]. AB - OBJECTIVE: To study nomal valuels of PI, RI and S/D in healthy male penile dorsum artery (DA) and cavernosal artery (CA). METHODS: 257 healthy mature men were divided into 5 groups by age. Group 1: <30, n=65; Group 2: 30-39, n=83; Group 3: 40-49, n=61; Group 4: 50-59, n=38; Group 5: > or = 60, n=10. Hibateral PI, RI and S/D values of penile dorsum artery and cavernosal artery were examined by Logidop(r)2 Type Digital Doppler Ultrasonography. RESULTS: There were no significant difference for PI, RI and S/D of penile dorsum artery and cavernosal artery not only in personal left and right artery but also in different age groups. Normal values advised: (1)LDA:PI 1.43-3.43, RI 0.72-0.92, S/D 2.68-10.56. (2)RDA PI 1.47-3.47, RI 0.73-0.93, S/D 3.27-10.09. (3)LCA:PI 1.49-3.21, RI 0.74 0.90, S/D 3.17-9.55. (4)RCA:PI 1.93-3.27, RI 0.72-0.90, S/D 3.22-9.42. CONCLUSION: Doppler ultrasonography is a favorable method in filtering penile arterial function. PMID- 16259365 TI - [Application of MSCT and post-processing images to fractures of nasal bone in forensic identification]. AB - OBJECTIVE: To evaluate the application of MSCT and post-processing images to fractures of nasal bone in forensic identification. METHODS: 134 cases were examined by thin slice scanning with MSCT and all of the data were sent to workstation for MPR and SSD. The result of MSCT was compared with that of X-ray. RESULTS: There are 55 (41.04%) cases of linear fracture, 46 (34.33%) cases of comminuted fracture, 27 (20.15%) cases of depressed fracture and 6 (4.48%) cases of no fracture in this sample. With X-ray or CR, 48 (35.82%) cases were misdiagnosed or underdiagnosed. 133 (99.25%) cases were confirmed by MSCT. Significance difference was found between X-ray and MSCT (chi2= 45.0816, P<0.001). CONCLUSION: MSCT and post-processing images might be the chief evidence for nasal fractures in forensic identification. PMID- 16259366 TI - [Anatermic separated epiphysis or avulsion fracture: image observation]. AB - OBJECTIVE: To explore an approach to distinguish avulsion fracture of articular portion from anatomic separated epiphysis. METHODS: 21 cases with suspicioned avulsion fracture of articular portion of tubular bone were reviewed. The cortical continuity of separate small bone and contiguous bone portion was investigated. The swell of soft tissue around these small bones was observed simultaneously. Then the configuration of the fracture was researched by injury mechanics. RESULTS: Continued bone cortex between separate small bone and corresponding bone was seen in 15 cases, so they were considered as a separate epiphysis. Sharp linear fracture between the separate small bone and corresponding bone were seen in 6 cases. The swell of soft tissue around the separate small bone was observed in all cases, so they were considered as a avulsion fracture. CONCLUSION: The meticulous investigation of separated small bone at the articular portion by image observation is of important value for distinguishing avulsion fracture from anatomic separate epiphysis. PMID- 16259367 TI - [Genetic polymorphism analysis of 6 STR loci on the X chromosome in Xi'an Han population]. AB - OBJECTIVE: To investigate the alleles and genotypes frequency of 6 short tandem repeat (STR) loci (DXS8378, DXS7132, DXS6789, DXS101, HPRTB and DXS7423) on the X chromosome in Han population. METHODS: The six X-chromosome STR loci were analyzed by PCR following polyacylamide gel electrophoresis and silver stain. RESULTS: Among 120 females from Xi'an Han population, the number of alleles in the 6 loci (DXS8378, DXS7132, DXS6789, DXS101, HPRTB and DXS7423) were 5, 6, 11, 10, 8, and 4 respectively; the number of genotypes in the 6 loci were 10, 17, 29, 32, 22, and 7 respectively; Exact tests demonstrated genotype frequencies in females had no departure from Hardy-Weinberg equilibrium. CONCLUSION: The six X chromosome STR loci are appropriate for individual identification, paternity testing involving a female child and for study on related disease. PMID- 16259368 TI - [Genetic polymorphism of DXS7423, DXS7424, DXS7132, DXS7133, DXS6804 loci on X chromosome in Ewenke population]. AB - OBJECTIVE: To study the genetic polymorphism of five STR loci on X-chromosome in 98 Ewenke population of China and establish the related genetic database. METHODS: The sequence variation of DXS7423, DXS7424, DXS7132, DXS7133, DXS6804 loci on X-chromosome were analyzed by PCR following PAG electrophoresis and silver staining. RESULTS: The genetic data of the 5 X-chromosome STRs in Ewenke ethnic group of China is in accordance with Hard-Weinberg equilibrium. CONCLUSION: Five X-STR loci could be used in forensic identification and population genetic research. PMID- 16259369 TI - [Species identification of biological samples by amplifying intron 8 of the TP53 gene]. AB - OBJECTIVE: To identify the species of biological samples by amplifying intron 8 of the TP53 gene. METHODS: Collected the bloodstains or muscle tissues of 14 kinds of common animals and human. DNA were quantified after extraction. The PCR amplification products were analyzed by PAGE and silver staining. RESULTS: The human and monkey have one amplification band of 460 bp while the eel, fish, frog, duck, rabbit, cat, mouse, cavy, pig, ox and sheep could amplify different unspecial bands. No amplification products were found in chicken and dog. CONCLUSION: The method of species identification by amplifying intron 8 of the TP53 gene is simple and sensitive. PMID- 16259370 TI - [Diversity of paternity index with different allele frequencies]. AB - OBJECTIVE: To evaluate the diversity of combined paternity index (CPI) of multiple STR loci when different population allele frequencies was used to calculate the paternity index. METHODS: CPI of 13 CODIS (combined DNA index system) loci for 108 trio cases and 108 duo cases selected randomly were calculated by using five Chinese Han population allele frequencies, respectively. RESULTS: The CPI range for trio cases and duo cases were 2077.63-50897711626.46 and 25.12-2998685141, respectively. When different population allele frequencies were applied to the same case, the ratio of maximum CPI and minimum CPI, which was more than 100, for trio cases and duo cases were 20 cases (19.52%) and 13 cases (12.04%), respectively. CONCLUSION: The variation of CPI value of the CODIS loci was obvious with different allele frequencies. To prevent the error causing by uncertain allele frequencies, a conservative CPI value should be calculated in paternity testing. PMID- 16259371 TI - [Study of rating scale of mentally prisoner's competency to serve a sentence]. AB - OBJECTIVE: To create an instrument to determine the mental prisoners' competency to serve a sentence, which is according with the Chinese legal system. METHODS: Integrating the Chinese criminal jurisprudence and the authors' forensic psychiatric experience, the research team created an instrument which called Competency to serve a sentence Rating scale firstly, then used the instrument retrospectively, in the end the validity and reliability of the instrument were inspected and, through an diagnostic test, the feasibility of the instrument was evaluated. RESULTS: Homogeneity reliability of the instrument is 0.8779, the correspondence of the conclusion between the instrument and the expertise is 0.909, except the positive likelihood ratio is 0.0683, the other diagnostic index are better. CONCLUSION: The Competency to serve a sentence Rating Scale is feasible. PMID- 16259372 TI - [The method and new advance in diagnosing erectile dysfunction]. AB - Recently, we have a further understanding on the pathogeny of erectile dysfunction, and with the application of so many new technologies, such as color Doppler ultrasound, electrical impedance, evoked potential, sensor etc. The diagnosis of ED becomes more and more scientific and convenient. Today we make a review on all kinds of diagnosing methods in order to recognise it more clearly and to provide some more precise and practical way for forensic evaluation in future. PMID- 16259373 TI - [Practice value of whole genome amplification technology to be used in forensic science and analysis of its result]. AB - Genetic analysis from forensic microsamples is a urgent, difficult task in forensic science, because it is frequently limited by the amount of specimen available in forensic practice, much effort has been carried out to resolve this difficulty. Whole genome amplification (WGA) technology, which was developing quickly in these years, has been thought to be a powerful, reliable and efficient strategy in analysis of minute amount DNA on many fields. In this review, we discuss its application in forensic science. PMID- 16259374 TI - [The expression of COX-1 and COX-2 following brain injuries]. AB - Cyclooxygenase (COX) is the obligate, rate-limiting enzyme for the conversion of arachidonic acid into prostaglandins, which mediate mitogenesis, apoptosis, angiogenesis, blood flow, secondary injury, and inflammation. COX is consist of 2 subtypes: COX-1 and COX-2. In recent years, there are a number of lines of evidence that COX-1 and COX-2 play a important in role brain injuries. PMID- 16259375 TI - In vitro application of endotoxin enhances nitric oxide production in thoracic aortas from Mg-deficient rats. AB - Since endotoxin-induced vascular hyporeactivity to phenylephrine is enhanced in Mg-deficient rats, this study was designed to determine whether endotoxin directly enhances nitric oxide (NO) production in thoracic aortas isolated from Mg-deficient rats in vitro. Thoracic aortas isolated from Mg-deficient and control rats were cultured for 6 h with or without endotoxin (LPS). LPS (0.01-1.0 microg) increased NO production in a concentration-dependent manner. NO production in the presence of 0.1 and 1.0 microg/mL LPS was significantly higher in Mg-deficient rat aortas compared to aortas from control rats. The enhanced NO production was not significantly affected by endothelium-denudation. LPS stimulated NO production was fully inhibited by a selective iNOS inhibitor, 1400W (0.1, 1.0 microM), in control rat aortas, but in Mg-deficient rat aortas inhibition by 1400W was only partial. A similar inhibitory effect was observed with anti-CD14 and anti-TLR4 antibodies. These results suggest that endotoxin enhances NO production in Mg-deficient rat aortas directly, and that endotoxin receptors might, at least in part, contribute to this enhancement. PMID- 16259376 TI - Reduction of perifusate magnesium alters inotropic response of papillary muscle to ion channel modulators. AB - Magnesium has a significant role in the regulation of ion transport. Marginal deficiency of Mg can therefore affect myocardial excitability and contractility. This study was taken up with the objective of examining the inotropic response of the myocardium to variation in extracellular [Mg]o and identifying the ion channels and pumps mediating the inotropic changes. Electrically stimulated rat papillary muscle was used as the experimental model and mechanical changes were recorded using a physiograph. Channel specific antagonists were used to identify the channels mediating the functional changes. Diastolic Ca2+ levels were determined in isolated myocytes by the ratiometric method using the fluorescent indicator Fura2-AM. A negative association was observed between the level of [Mg]o and force of contraction, with a peak at 0.48 mM Mg. The force of contraction in Mg deficient medium (0.48 mM) was 158% of control (1.2 mM Mg) (p < 0.001). Inotropic response to the L-type channel antagonist (verapamil-1 microm) and NaK ATPase inhibitor (Ouabain-0.3 mM) was augmented in Mg deficiency (p < 0.005), indicating activation of the channel and the pump. The response to T-type channel inhibitor (NiCl2-40 microM) was attenuated in Mg deficiency (p < 0.05). The response to the sarcoplasmic reticular Ca pump inhibitor (caffeine-10 mM) and the SR Ca2+ release channel inhibitor (ryanodine-1 microM) were not significantly affected by Mg deficiency. Diastolic level of Ca2+ increased with a decrease in Mg (p < 0.05). The observations of the study lead to the conclusion that the positive inotropic response in Mg deficiency is mediated by an increase in basal Ca2+ combined with Ca-induced-Ca release consequent to Ca2+ influx through L-type Ca channel. Variation in sensitivity to Ca channel blockers and NaK ATPase inhibitor in Mg deficiency can have pharmacological implications. PMID- 16259378 TI - Na+/Mg2+ antiport in erythrocytes of spontaneously hypertensive rats: role of Mg2+ in the pathogenesis of hypertension. AB - Total Mg2+ content in plasma and erythrocytes did not significantly differ between WKY and SHR. Mg2+ efflux via Na+/Mg2+ antiport was 10% lower in non Mg(2+)-loaded erythrocytes of SHR than in WKY, and 16% lower in Mg(2+)-loaded erythrocytes of SHR. The activation of Na+/Mg2+ antiport in erythrocytes by Cl-, as tested by substitution of Cl- with SCN-, and the regulation of Na+/Mg2+ antiport by protein kinases, as tested by PMA and staurosporine, showed no differences between WKY and SHR. The reduction of Na+/Mg2+ antiport was explained by a reduction in the number of Na+/Mg2+ antiporter molecules in SHR erythrocytes. Mg2+ efflux in KCl medium by K+/Mg2+ antiport via the unspecific choline exchanger was not significantly reduced in SHR and was equally affected by PMA and staurosporine in WKY and SHR. An explanation for some controversial results, unchanged or reduced concentration of Mg2+ in serum, total Mg2+ and free Mg2+ in erythrocytes of SHR and patients with essential hypertension was proposed. The role of Na+/Mg2+ antiport and [Mg2+]i in the pathogenesis of experimental and clinical hypertension was discussed. PMID- 16259377 TI - Effects of magnesium sulfate on spinal cord tissue lactate and malondialdehyde levels after spinal cord trauma. AB - OBJECTIVE: In the present study, the effects of magnesium sulfate (MgSO4) on tissue lactate and malondialdehyde (MDA) levels after spinal cord trauma (SCT) in rabbits were studied. SUBJECTS: Thirty New Zeland rabbits. Interventions. The rabbits were divided equally into three groups: group I was the sham- operated group, group II suffered from SCT but received no treatment, group III was given a dose of 100 mg/kg of magnesium sulfate intravenously at 5th minute after SCT. MEASUREMENTS. The lactate and MDA levels were measured in contused spinal cord tissue at 60 minutes after SCT. There was a significant increase of lactate and MDA levels in group II (p < 0.05) when compared with groups I and III, and a significant increase in the level of MDA in group III compared with group I, and also a significant decrease compared with group II, which was the trauma group without treatment (p < 0.05). CONCLUSION: The findings of this study showed that magnesium sulfate can attenuate the increase of tissue MDA and supply a normalization of lactate levels following SCT which may be related to the neuroprotective effects of (MgSO4). PMID- 16259379 TI - Magnesium chloride or magnesium sulfate: a genuine question. AB - MgSO4 is routinely used in therapeutics despite its toxicity. The aim of the present review was to compare MgSO4 and MgCl2 effects in order to answer the question whether MgSO4 could be or not replaced by MgCl2. Considering that the two salts have both similar and proper effects, a clear-cut conclusion is not easy to draw. However, choosing MgCl2 seems advisable because of its more interesting clinical and pharmacological effects and its lower tissue toxicity as compared to MgSO4. PMID- 16259380 TI - Self-construals and values in different cultural and socioeconomic contexts. AB - In this study the authors investigated (a) how individuational and relational self-orientations, as well as self-directed and other-directed values, are related to one another, and (b) how these self- and value orientations differ across 2 cultural (i.e., 422 Turkish and 441 American university students) and 2 socioeconomic status (SES) groups (i.e., 186 lower SES and 167 upper SES Turkish high school students). Across cross-cultural and SES groups, individuational and relational self-orientations appeared to be not opposite but distinct orientations, as predicted by the Balanced Integration-Differentiation (BID) model (E. O. Imamoglu, 2003). Furthermore, both Turkish and American students with similar self-construal types, as suggested by the BID model, showed similar value orientations, pointing to both cross-cultural similarities and within cultural diversity. Individuational and relational self-orientations showed weak to moderate associations with the respective value domains of self-directedness and other-directedness, which seemed to represent separate but somewhat positively correlated orientations. In both cross-cultural and SES groups, students tended to be high in both relational and individuational self orientations; those trends were particularly strong among the Turkish and American women compared with men and among the upper SES Turkish adolescents compared with lower SES adolescents. Results are discussed as contesting the assumptions that regard the individuational and relational orientations as opposites and as supporting the search for invariant aspects of psychological functioning across contexts. PMID- 16259381 TI - The paradox of promoting creativity in the Asian classroom: an empirical investigation. AB - To shed light on the paradox of promoting creativity in the Asian classroom, the authors conducted 3 studies. The 1st study found that novice teachers classified student behaviors as desirable but uncreative (DBU) versus creative but undesirable (CBU). The 2nd study found that conservative-autocratic teachers were more likely to encourage DBU behaviors in class, whereas liberal-democratic teachers were more likely to encourage CBU behaviors in class. The 3rd study found that cultural individualism-collectivism had a positive impact on liberal democratic teaching attitude but a negative impact on conservative-autocratic teaching attitude. In turn, liberal-democratic teaching attitude had a positive impact on the tendency to promote CBU behaviors, whereas conservative-autocratic teaching attitude had a positive impact on the tendency to promote DBU behaviors. PMID- 16259382 TI - When ideals are too "far off': physical self-ideal discrepancy and body dissatisfaction in Japan. AB - There is much evidence that young Japanese adults manifest relatively low body esteem-a phenomenon often explained as an outcome of modesty or limited need for self-enhancement. The author sought to identify additional determinants of this phenomenon and to explain its sources by examining the relationship between several presumed factors and level of body satisfaction rated by 263 Japanese students. The findings confirm earlier evidence of low body satisfaction among young Japanese adults and indicate that it is predicted mainly by a discrepancy between perceptions of the actual body and the ideal, self-esteem, and a predisposition to interpersonal phobia. Although some of the factors revealed are relevant to other cultures as well, the relatively wide discrepancy between self and ideal body and predisposition to interpersonal phobia may characterize young Japanese adults in particular and stem from a specific historical background. Overall, the results suggest that culture has a significant role in shaping collective body images in Japan and that a broader conceptualization of physical self-ideal discrepancy may better explain divergent levels of body satisfaction across cultures. PMID- 16259383 TI - Making "someday" happen--the years of living dangerously. PMID- 16259384 TI - Retirement and un-retirement in the 21st century. PMID- 16259385 TI - Physicians: gender-neutral professional responsibilities. PMID- 16259386 TI - Is Medicaid relevant to your practice? Take a survey-let us know. PMID- 16259387 TI - Highlights of the 2005 annual American Medical Association meeting. PMID- 16259388 TI - Health care coverage should be amendment XXVIII--not a business commodity. PMID- 16259389 TI - Responding to an emergency in and out of the office. AB - Moral duties aside, a physician generally has no legal liability to a stranger in a medical emergency if the physician decides to do nothing so long as the physician is not under a pre-existing duty or has created a risk of harm to such stranger. Missouri, like any other state, has a Good Samaritan statute specifically designed to encourage medical care providers to offer assistance to injured persons at the scene of an emergency or accident. Although, the statute on its face draws the line between ordinary negligence and gross negligence in extending the immunity to any qualified rescuer who provides emergency medical care without compensation at the scene of emergency, no Missouri case law has recognized "gross negligence" as a cause of action at common law. The practical effect is that it is unlikely any qualified rescuer will be held liable for a victim's injuries when the care was rendered in good faith during emergency and without compensation, however negligently, unless there is willful or wanton misconduct on the part of such rescuer. PMID- 16259390 TI - Legalizing imported medicines will increase health and safety risks to American patients. PMID- 16259392 TI - Physician conduct in tort litigation. PMID- 16259391 TI - Mentoring young minds. PMID- 16259393 TI - Update on anesthesiology and critcal care medicine. PMID- 16259394 TI - Bacteriocidal activity of lizard and mouse serum for Borrelia lonestari, putative agent of a Lyme-like illness (AKA STARI or Masters disease) in Missouri. AB - OBJECTIVE: To determine responses of Borrelia lonestari and Borrelia burgdorferi to Eastern Fence lizard (Sceloporus undulatus) and Swiss-Webster mouse (Mus musculus) sera. RESULTS: Lizard sera lysed both Borrelia lonestari and Borrelia burgdorferi. Mouse sera lysed only Borrelia lonestari. CONCLUSIONS: Borrelia lonestari and Borrelia burgdorferi spirochetes did not survive exposure to lizard sera. Mice are reservoirs for Borrelia burgdorferi but may not be a Borrelia lonestari reservoir because spirochetes did not survive exposure to mouse sera. PMID- 16259395 TI - Evolving concepts in pediatric anesthesia. AB - The cuffed endotracheal tube and laryngeal mask airway are very useful in the airway management of children. Safely anesthetizing and recovering a child with obstructive sleep apnea requires an understanding of both the altered anatomy and physiology of the patient. The addition of clonidine in caudal blocks prolongs pain relief. PMID- 16259396 TI - Virtual reality and medicine--from the cockpit to the operating room: are we there yet? AB - Teaching medicine to medical students, physicians in training and nurses is a challenging task that has remained unchanged for decades. The airline industry has achieved a great deal of safety and quality in a technically challenging environment. Many believe that their outstanding achievement is due to team training and crew resource management using simulators and dedicated training programs. Many experts in the medical profession believe that adopting the same strategies in teaching medical students and trainees could achieve significant reductions in medical errors and improve the quality of patient care. This article explores the role of teaching medicine using virtual reality in a multitude of medical specialties and outlines the use of simulation training at Saint Louis University. PMID- 16259398 TI - Perioperative transesophageal echocardiography. AB - The first reported use of the transesophageal route to record the thoracic aortic flow using continuous wave Doppler was described by Side and Gosling in 1971. Since then, transesophageal echocardiography (TEE) has become a valuable tool in monitoring and diagnosis of heart function when indicated during surgery. Currently, anesthesiologists with competence in TEE play an indispensable role in perioperative care. This review will cover several of the important uses of TEE as both a monitor and diagnostic aid in anesthesiology. PMID- 16259397 TI - Update on chronic pain management. AB - Chronic pain remains a significant health problem in the United States. Old treatments are being modified, and new treatment options are being developed based on outcome studies. This article addresses some of the new concepts in conservative and invasive treatment options that are supported by outcome studies for the treatment of chronic pain patients. PMID- 16259399 TI - Acute respiratory distress syndrome update. AB - Respiratory failure and eventual ventilator dependence are hallmarks of the acute respiratory distress syndrome (ARDS). ARDS carries a significant mortality in all patient populations and the treatment strategies are largely supportive in nature. Despite years of clinical experience, ARDS continues to pose a substantial threat and no definitive therapy has yet to be described. We will present an overview of management options. PMID- 16259400 TI - Arthritis in Missouri: disability, quality of life and treatment. AB - This study estimates Missouri's arthritis and activity limitation burden using the Behavioral Risk Factor Surveillance System data. In 2001, approximately one third of Missouri adults reported physician-diagnosed arthritis and/or chronic joint symptoms, while over one-tenth reported an activity limitation from joint symptoms. These conditions are common among individuals with other chronic health problems and contribute to disability. The Missouri Arthritis and Osteoporosis Program provides self-management resources that can improve patient outcomes and quality of life. PMID- 16259401 TI - Pathology, prognosis, and prevention of heart failure. AB - Optimal management of patients with heart failure (HF) should be guided by thorough evaluation and use of functional classification and disease staging systems. Though the understanding of precise underlying pathophysiological mechanisms has increased dramatically in recent years, it is essential to continue the search for greater efficacy in preventing, controlling, and reversing this pathological state. Recent developments in the diagnosis and treatment of heart failure have improved prognosis in terms of both survival and morbidity due to re-hospitalization. The underutilization of medical and preventive therapies in patients with heart failure is a major public health issue, despite the recommendations of various evidence-based guide lines. Patient education and disease management programs have been shown to optimize HF management by bridging the gap between evidence-based medicine and clinical practice, thereby, reducing hospitalization rate. This paper, the first in a series of three papers, will identify the pathology, prognosis and prevention of heart failure, while the paper to follow will discuss modern methods of interventional management. PMID- 16259402 TI - [PolySomnographic findings in pickwickian syndrome]. PMID- 16259403 TI - [Psychophysiological basis of smells]. AB - There has been an increase in the use of psychophysiological techniques, especially event-related potentials (ERPs) to evaluate the effects of odor on the central nervous system. In the study of ERPs related to odor, there are two main methods of the odor presentation: (1) to investigate the olfactory ERP (OERP) or chemosensory ERP (CSERP), that is to calculate electrophysiological responses to odor stimulation as a trigger; (2) to compare ERP of mental functioning during the odor exposed condition with that of no odor exposure. The amplitude of contingent negative variation (CNV) varies with the odorant being present, when the odor is presented as a trigger and when it is administrated during a task. It is suggested that CNV changes depend on not only the odorant but the anticipation, expectation and emotion of the subject. The latency of the N1 component becomes shorter with increasing concentrations of odors. The N1 amplitude does not depend on the odor concentration while the amplitude of the auditory N1 strongly depends on the stimulus intensity. There is only one report that the olfactory mismatch negativity (MMN) is present in the CSERP. On the other hand, auditory MMN is not affected by odor administration. In relation to the P3 component, rare odors evoked a larger amplitude in contrast to frequent odors, as well as P3 evoked by other modalities. These ERP studies with odors are expected to be applied to such clinical settings as the differentiation between the anosmic patients and normosmic persons, the functional evaluation of patients with brain tumors, the earlier detection of dementia, and the objective evaluation of aromatherapy. PMID- 16259404 TI - [Effect of acupuncture treatment for a patient with severe axial dystonia appearing during treatment for schizophrenia]. AB - We performed acupuncture treatment on first consultation for a female 28-year-old patient with severe axial dystonia, causing involuntary movement and abnormal posture of the neck and body, which had developed during treatment for schizophrenia. Involuntary movement involving elevation of the right shoulder began to occur in October X-1. Drugs were prescribed by her doctor, but her involuntary movement worsened and spread to the whole body. Thereafter, she began receiving acupuncture treatment at the out patient clinic for dystonia at the Kansai Medical College Hospital in July X. Involuntary movements of her neck involved repeated left lateral bending or a rigidly straight posture while sitting and standing. Her neck also showed a left lateral bend and right rotation. Her body showed a left lateral bend and right shoulder elevation. The neck problems in this case were induced by a hypertonicity of the left sternocleidomastoid (SCM), which caused the left lateral bending and right rotation of the neck. Problems in her body involved left lateral bending due to hypotonicity of the left abdominal muscle and hypotonicity of the left back muscles, which were unable to control the left lateral bending of the body. The right shoulder elevation was caused by a hypertonicity of the right trapezius and this was another of her problems. Acupuncture treatments were given using a penetrating needle method. The treatment points were left LI4 to decrease the hypertonicity of the left SCM, left ST41 to increase the hypotonicity of the left abdominal muscles, right BL60 to increase the hypotonicity of the right back muscles and right TE5 to decrease the hypertonicity of the right trapezius. At the initial stage of acupuncture treatment, the patient was not able to attend the hospital regularly enough to obtain sufficient improvement by acupuncture. In December X+1, she started to receive acupuncture treatment weekly, and the posture of the neck and body improved. In May X+3, her neck and body postures remained erect while sitting and she did not show involuntary movement. For problems of dystonia, we perform acupuncture treatment, using meridian and acupressure points selected based on the oriental medicine system, and we achieved improvement of symptoms in this case. The patient also achieved improved stability with regard to the symptoms of schizophrenia. It is suggested that acupuncture treatment has had a positive effect on tardive dystonia including axial dystonia. PMID- 16259405 TI - [Atypical antipsychotic-associated impaired glucose tolerance]. AB - Some reports of impaired glucose tolerance associated with olanzapine (OLZ) treatment have been published before OLZ was marketed in Japan. In Japan, we have been prohibited from using OLZ for patients with diabetes mellitus, since several cases with OLZ-associated impaired glucose tolerance including two deaths from diabetic coma have been reported. Here, we report four cases of OLZ-associated impaired glucose tolerance and review the points to consider in treatment with OLZ. Of our four cases, three cases were new-onset (non diabetes mellitus cases) and the other case was a diabetes mellitus-existent (diabetes mellitus case). In the non DM cases, the time to the onset of impaired glucose tolerance after initiating treatment with OLZ was 8-9 months, and the impaired glucose tolerance immediately improved after discontinuing treatment with OLZ and initiating treatment for diabetes mellitus. Therefore, it is necessary to continue long-term monitoring of the parameters of glucose metabolism for all patients treated with OLZ. Should impaired glucose tolerance develop during treatment with OLZ, treatment with OLZ should be discontinued immediately and treatment for diabetes mellitus should be started if necessary. Although the condition of diabetes mellitus was stable befor initiating treatment with OLZ in the DM case, hyperglycemia developed immediately after initiating treatment with OLZ and the condition remained unstable even after early treatment for diabetes mellitus. Therefore, it is necessary to check for a previous history of diabetes mellitus and hyperglycemia befor initiating treatment with OLZ. Correlations between weight gain and occurrence of impaired glucose tolerance are not clear, so it is necessary to monitor the occurrence of impaired glucose tolerance even in cases without weight gain. PMID- 16259406 TI - Standards for psychiatric residency training and certification: issues for institutions and program graduates. PMID- 16259407 TI - [My clinical experience in interaction with patients]. AB - In clinical practice, many kinds of stories are told by patients, stories such as those about distress associated with illness, those about their interaction with the people surrounding and those about the medical experiences. Stories are never fixed. They are always revised. Several dimensional stories are interrelated and affect one another, and the modification of any influences the others. The stories told by patients are coded and interpreted so as to fit the doctors' paradigm. The coding and interpretation are sometimes willingly accepted by patients for the purpose of being recognized as patients by the community, or because the patients need to close their eyes to their real mind state. In other cases they are rejected because patients perceive the application of paradigms either as doctors' indifference about patients' stories or even as a kind of violence. There are various kinds of paradigms. The choice of a paradigm is determined not only by the patients' disease nature but the personal history of individual doctor and the academic group which the doctor is belonging to. So the paradigm has always has the cultural aspect. Through the interaction between patients' stories and doctors' explanation model based on the paradigms, new stories start developing. How the application of the doctors' explanation models impact each patient should be intuitively considered beforehand, keeping in mind the patients' past histories, the situation the patients are now put in, past experiences with medical services. This is because the medical explanations offered to the patients could change their experience of illnesses in various ways, sometimes making them more distressful. To indorse my opinions, I extracted some pieces from my description about two patients. PMID- 16259408 TI - [Swiss scrapie surveillance. I. Clinical aspects of neurological diseases in sheep and goats]. AB - Small ruminants infected with scrapie show a large range of often unspecific clinical symptoms. The most-often described signs, locomotion, sensibility and behavioural disorders and emaciation, rarely occur together, and cases have been described in which only one of those signs was detectable.Thus, formulating a well-circumscribed definition of a clinical suspect case is difficult. Most animals with CNS-effecting diseases such as listeriosis, polioencephalomacia, cerebrospinal nematidiasis and enterotoxemia will, in a thorough neurological examination, show at least some scrapie-like symptoms. Among the 22 neurological field cases examined in this study, a goat with cerebral gliomatosis and hair lice showed the closest similarity to clinical scrapie. The unilateral deficiency of the cerebral nerves has potential as an clinical exclusion criterion for scrapie. However, the laboratory confirmation--or exclusion--of scrapie remains important. It thus needs to be realized that a consistent and thorough examination of neurologically diseased small ruminants (including fallen stock) is the backbone of a good surveillance system for these diseases. This should be a motivation for submitting adult sheep and goats for neuropathological examination. PMID- 16259409 TI - [Swiss scrapie surveillance. II. Epidemiologic aspects of the detection of neurological diseases in sheep and goats]. AB - Monitoring of transmissible spongiform encephalopathy (TSE) in Swiss sheep and goats is based on the examination of animals from different sources. In this study, frequencies and proportions of the different diagnoses were compared between routinely submitted sheep and goats, notified scrapie suspects as well as fallen stock. Meningitis/ encephalitis cases were significantly more frequent (OR = 2.2) in the scrapie suspect group when compared to the routine submissions. Metabolic-toxic encephalopathy was seen more frequently within the fallen stock. Rare neurological diagnoses were more frequent among scrapie suspects and routine submissions when compared to fallen stock. Listeriosis was diagnosed equally frequent among the scrapie suspects and routine submissions but less frequent in fallen stock. Scrapie prevalence among the fallen stock and the routine submissions was 0 (zero), with 95% certainty that prevalence is < 1%. The examined animals are representative for most of the Swiss regions with considerable sheep and goat production. Continuation of the detailed neuropathological examination of small ruminants from these three groups, substituted by actively testing a sufficiently large sample of fallen stock and possibly also healthy-slaughtered adult sheep and goats for transmissible spongiform encephalopathies would ensure a good surveillance within the small ruminant population. PMID- 16259410 TI - [Abortion in sheep: epidemic Salmonella abortusovis outbreak 2005 in Switzerland]. AB - In spring 2005, the outbreak of contagious abortion caused by Salmonella Abortusovis in 6 sheep flocks in Switzerland led to considerable economic losses. The Swiss small ruminant health service (BGK) evaluated this case. The aim was to identify the source of the epidemic in order to avoid further spread of infection and to evaluate the possibility of using vaccination. Moreover, a strategy for prevention of future outbreaks was developed. This article aims to increase disease awareness of food animal practitioners for Salmonella Abortusovis abortion in sheep. PMID- 16259411 TI - Astrocytoma in a chamois. AB - Astrocytomas represent the most common cerebral tumors in humans and in animals, and the fibrillary cytological subtype is the most frequently observed. In this report and for the first time, a thalamic astrocytoma is described in a chamois showing depressed mentation, pleurothotonus and circling to the right side. PMID- 16259412 TI - What will you do for your profession? PMID- 16259413 TI - Joy in dentistry. PMID- 16259414 TI - Compliance with the code: an ethical obligation. PMID- 16259415 TI - Support for the chemically dependent dentist. PMID- 16259416 TI - Identifying drug problems in dentistry. PMID- 16259417 TI - Diagnosis and treatment of chemical dependency in professionals. AB - Accurate diagnosis of chemical dependence is possible with careful evaluation of the patient following the preceding general guidelines. While history is as critical in this field as it is in every other area of medicine or dentistry, we must remain cognizant of resistance in accurate history taking, since the patient has a need to keep the disease hidden. The need for corroborating history through interviews with significant others, spouses or other family members is critical. We all must be aware of the impact of social stigma on our willingness to make the diagnosis, particularly in colleagues, to ensure that we are willing to consider the diagnosis when necessary. Since physicians, dentists, nurses and the other professionals are at least as likely to have chemical dependence as the rest of the population, we must also be willing to identify and discuss with our colleagues issues of chemical dependence. This requires a great deal of courage, since it is often more difficult to confront a friend or colleague with evidence of chemical abuse or dependence. Yet a true friend is willing to confront, particularly when a colleague's family, well-being and professional life are at stake. In the case of chemical dependency, we must be our brothers' and sisters' keeper. PMID- 16259420 TI - Personnel files: practical guidelines for your office. PMID- 16259421 TI - Criteria and air-toxic emissions from in-use automobiles in the National Low Emission Vehicle program. AB - The U.S. Environmental Protection Agency (EPA) implemented a program to identify tailpipe emissions of criteria and air-toxic contaminants from in-use, light-duty low-emission vehicles (LEVs). EPA recruited 25 LEVs in 2002 and measured emissions on a chassis dynamometer using the cold-start urban dynamometer driving schedule of the Federal Test Procedure. The emissions measured included regulated pollutants, particulate matter, speciated hydrocarbon compounds, and carbonyl compounds. The results provided a comparison of emissions from real-world LEVs with emission standards for criteria and air-toxic compounds. Emission measurements indicated that a portion of the in-use fleet tested exceeded standards for the criteria gases. Real-time regulated and speciated hydrocarbon measurements demonstrated that the majority of emissions occurred during the initial phases of the cold-start portion of the urban dynamometer driving schedule. Overall, the study provided updated emission factor data for real world, in-use operation of LEVs for improved emissions modeling and mobile source inventory development. PMID- 16259422 TI - Combustion characteristics of spent catalyst and paper sludge in an internally circulating fluidized-bed combustor. AB - Combustion of spent vacuum residue hydrodesulfurization catalyst and incineration of paper sludge were carried out in thermo-gravimetric analyzer and an internally circulating fluidized-bed (ICFB) reactor. From the thermo-gravimetric analyzer differential thermo-gravimetric curves, the pre-exponential factors and activation energies are determined at the divided temperature regions, and the thermo-gravimetric analysis patterns can be predicted by the kinetic equations. The effects of bed temperature, gas velocity in the draft tube and annulus, solid circulation rate, and waste feed rate on combustion efficiency of the wastes have been determined in an ICFB from the experiments and the model studies. The ICFB combustor exhibits uniform temperature distribution along the bed height with high combustion efficiency (>90%). The combustion efficiency increases with increasing reaction temperature, gas velocity in the annulus region, and solid circulation rate and decreases with increasing waste feed rate and gas velocity in the draft tube. The simulated data from the kinetic equation and the hydrodynamic models predict the experimental data reasonably well. PMID- 16259423 TI - Inhalation of concentrated ambient particulate matter near a heavily trafficked road stimulates antigen-induced airway responses in mice. AB - The goal of this study was to test the following hypotheses: (1) exposure to mobile emissions from mobile sources close to a heavily trafficked roadway will exacerbate airway inflammation and allergic airway responses in a sensitized mouse model, and (2) the magnitude of allergic airway disease responses will decrease with increasing distance from the roadway. A particle concentrator and a mobile exposure facility were used to expose ovalbumin (OVA)-sensitized BALB/c mice to purified air and concentrated fine and concentrated ultrafine ambient particles at 50 m and 150 m downwind from a roadway that was heavily impacted by emissions from heavy duty diesel-powered vehicles. After exposure, we assessed interleukin (IL)-5, IL-13, OVA-specific immunoglobulin E, OVA-specific immunoglobulin G1, and eosinophil influx as biomarkers of allergic responses and numbers of polymorphonuclear leukocytes as a marker of inflammation. The study was performed over a two-year period, and there were differences in the concentrations and compositions of ambient particulate matter across those years that could have influenced our results. However, averaged over the two-year period, exposure to concentrated ambient particles (CAPs) increased the biomarkers associated with airway allergies (IL-5, immunoglobulin E, immunoglobulin G1 and eosinophils). In addition, mice exposed to CAPs 50 m downwind of the roadway had, on the average, greater allergic responses and showed greater indications of inflammation than did mice exposed to CAPs 150 m downwind. This study is consistent with the hypothesis that exposure to CAPs close to a heavily trafficked roadway influenced allergic airway responses. PMID- 16259425 TI - Improving urban air quality in China: Beijing case study. AB - China is undergoing rapid urbanization because of unprecedented economic growth. As a result, many cities suffer from air pollution. Two-thirds of China's cities have not attained the ambient air quality standards applicable to urban residential areas (Grade II). Particulate matter (PM), rather than sulfur dioxide (SO2), is the major pollutant reflecting the shift from coal burning to mixed source pollution. In 2002, 63.2 and 22.4% of the monitored cities have PM and SO2 concentrations exceeding the Grade II standard, respectively. Nitrogen oxides (NOx) concentration kept a relatively stable level near the Grade II standard in the last decade and had an increasing potential in recent years because of the rapid motorization. In general, the air pollutants emission did not increase as quickly as the economic growth and energy consumption, and air quality in Chinese cities has improved to some extent. Beijing, a typical representative of rapidly developing cities, is an example to illustrate the possible options for urban air pollution control. Beijing's case provides hope that the challenges associated with improving air quality can be met during a period of explosive development and motorization. PMID- 16259424 TI - Hazardous air pollutants from mobile sources in the Metropolitan Area of Mexico City. AB - Environmental agencies are currently in the process of implementing a new air management program, which includes the improvement of fuel quality. In this work, exhaust emissions data and estimated relative risk for various fuels testing in use vehicles, equipped with three different exhaust emission control technologies, are presented. Aromatics, sulfur, and olefins contents; type of oxygenated compound; and Reid vapor pressure were varied. The aim also includes calculating the ozone (O3) forming potential and a relative cancer risk of emissions from current and formulated gasoline blends in Mexico. The proposed gasoline decreases carbon monoxide, total hydrocarbons (THC), and nitrogen oxides emissions by 18 and 14%, respectively, when compared with gasoline sold in the rest of the country and within ozone nonattainment metropolitan areas in Mexico, respectively. PMID- 16259426 TI - Predictions by the Odor From Feedlots, Setback Estimation Tool (OFFSET) compared with observations by neighborhood monitors. AB - A comparison between predicted and observed odor intensities at 20 neighborhood residences in the vicinity of seven various livestock farms in five different Minnesota counties was made to evaluate the Odor From Feedlots, Setback Estimation Tool (OFFSET) developed by the University of Minnesota. Observations by neighborhood monitors suggest that the OFFSET-predicted separation distances for annoyance-free frequencies of 99, 98, and 97% are large enough. The observations additionally indicate that predicted distances to obtain 94 and 91% annoyance-free frequencies may be large enough for some farms, but for other farms, greater distances may be needed. For two farms in the study, no significant difference between all of the observed and predicted intensities could be found. At four sites, a significant difference was found, and at three of these the difference was considerable. Odor emission rates used in the OFFSET model seem to describe the average emission fairly well for many odor sources, but improvement may be needed for some types of sources. Possible reasons for observations of annoying odor when not predicted include fluctuations in the odor emissions, wind speed fluctuations, topographic variation between sites, sensitivity differences by neighborhood monitors, and background emissions from other sources. PMID- 16259427 TI - Properties of pyrolytic chars and activated carbons derived from pilot-scale pyrolysis of used tires. AB - Used tires were pyrolyzed in a pilot-scale quasi-inert rotary kiln. Influences of variables, such as time, temperature, and agent flow, on the activation of obtained char were subsequently investigated in a laboratory-scale fixed bed. Mesoporous pores are found to be dominant in the pore structures of raw char. Brunauer-Emmett-Teller (BET) surfaces of activated chars increased linearly with carbon burnoff. The carbon burnoff of tire char achieved by carbon dioxide (CO2) under otherwise identical conditions was on average 75% of that achieved by steam, but their BET surfaces are almost the same. The proper activation greatly improved the aqueous adsorption of raw char, especially for small molecular adsorbates, for example, phenol from 6 to 51 mg/g. With increasing burnoff, phenol adsorption exhibited a first-stage linear increase followed by a rapid drop after 30% burnoff. Similarly, iodine adsorption first increased linearly, but it held as the burnoff exceeded 40%, which implied that the reduction of iodine adsorption due to decreasing micropores was partially made up by increasing mesopores. Both raw chars and activated chars showed appreciable adsorption capacity of methylene-blue comparable with that of commercial carbons. Thus, tire-derived activated carbons can be used as an excellent mesoporous adsorbent for larger molecular species. PMID- 16259428 TI - Vapor-phase and fine particulate matter concentrations of polycyclic aromatic hydrocarbons measured during the winter months in a northern Rocky Mountain urban airshed. AB - A fine particulate matter (PM2.5) sampling program was conducted in Missoula, MT, to investigate both the particle and vapor phases of PM2.5-associated polycyclic aromatic hydrocarbons (PAHs) found in a northern Rocky Mountain urban airshed. Twenty-four-hour samples were collected during the cold winter months of January through April 2002, when many of the more volatile organic components of PM2.5 were expected to be found in the condensed particle form. To meet analytical detection limits, each of the 12 individual sample days were aggregated into four total filter and polyurethane foam (PUF) samples, respectively, with each aggregate containing 3 sample days. Quartz filter (particle-phase PAHs) and PUF (vapor-phase PAHs) aggregates were analyzed separately for 18 individual PAHs and phenolics by gas chromatography/mass spectrometry. Results showed that 87% of the PM2.5-associated phenolics and PAHs measured in this study were found in the vapor phase. PM2.5-associated gas/particle partition coefficients (Kp,2.5) ranged from 0 for the lighter phenolics and PAHs to approximately 0.1 for some of the heavier PAHs, such as fluoranthene and pyrene. Calculating Kp,2.5 for the heaviest measured PAHs was not feasible because of low or undetectable concentrations in the vapor phases of these compounds. Phenolics and two-ringed and three-ringed PAHs were found almost exclusively in the vapor phase. Four ringed PAHs were distributed between the particle and vapor phases, with more mass measured in the vapor phase. Very little five-ringed and higher PAHs were measured from either the filter or PUF sampling medium. These results provide information on both the concentrations and different phases of PM2.5-associated PAHs measured during the winter months in a northern Rocky Mountain urban airshed, when concentrations of PM2.5 are generally at their highest compared with the rest of the year. PMID- 16259429 TI - Chemical mass balance model with fractionation for apportioning PM2.5: a test case for Los Angeles traffic sources. AB - Chemical mass balance receptor models (CMBs) use measured pollutant concentrations, along with source information, to apportion the contributions of primary sources to the measured concentrations. CMBs can be used to evaluate the accuracy of the emission inventories that underlie State Implementation Plan (SIP) modeling, by providing an allocation of emissions to individual source categories. CMBs, however, traditionally have not accounted for the chemical reaction and differential deposition or fractionation that occur between the source and receptor. This means that they have historically had severe limitations in apportioning secondary particulate matter (PM), which is an especially important component of fine PM (PM2.5). Stafford and Liljestrand developed a method to account for fractionation in CMBs using depletion factors based on a solution of the steady-state advection-dispersion equation, including gravitational settling, dry deposition, and first-order chemical reaction. PMID- 16259430 TI - Heterogeneous oxidation by ozone of naphthalene adsorbed at the air-water interface of micron-size water droplets. AB - The mass transfer of naphthalene vapor to water droplets in air was studied in the presence of ozone (O3) in the gas phase. A falling droplet reactor with water droplets of diameters 55, 91, and 182 microm was used for the study. O3 reacted with naphthalene at the air-water interface, thereby decreasing the mass transfer resistance and increasing the rate of uptake of naphthalene into the droplet. A Langmuir-Hinshelwood reaction mechanism at the air-water interface satisfactorily described the surface reaction. The first-order surface reaction rate constant, ks, increased with decreasing droplet size. Three organic intermediates were identified in the aqueous phase as a result of ozonation of naphthalene at the surface of the droplet indicating both peroxidic and nonperoxidic routes for ozonation. The presence of an organic carbon surrogate (fulvic acid) increased both the partition constant of naphthalene and the surface reaction rate of O3. The heterogeneous oxidation of naphthalene by O3 on the droplet was 15 times faster than the homogeneous oxidation by O3 in the bulk air phase, whereas it was only 0.08 times the homogeneous gas-phase oxidation by hydroxyl radicals under atmospheric conditions. PMID- 16259431 TI - Interactive analysis of waste recycling and energy recovery program in a small scale incinerator. AB - Conflicting goals affecting solid waste management are explored in this paper to find the best implementation of resource recovery with a small-scale waste-to energy process. Recycling paper and plastic material often leaves a shortage of thermal energy to support incineration that forces operators to supplement the process with auxiliary fuels. Although there are considerable profits to be made from material recovery, the increase of fuel usage causes conflict given that it is cost prohibitive. A series of trials performed on a small-scale 1.5-t/day incineration plant with a cyclone heat recovery system found that material recycling can impede performance. Experimental results are expressed as empirical regression formulas with regard to combustion temperature, energy transfer, and heat recovery. Process optimization is possible if the waste moisture content remains <30%. To test the robustness of the optimization analysis, a series of sensitivity analyses clarify the extent of material recycling needed with regard to plastic, paper, and metal. The experiments also test whether the moisture in the waste would decrease when recycling paper because of its exceptional capacity to absorb moisture. Results show that recycling paper is strongly recommended when the moisture content is >20%, whereas plastic recycling is not necessary at that moisture condition. Notably, plastic recovery reduces the heat needed to vaporize the water content of the solid waste, thus it is recommended only when the moisture content is <10%. For above-normal incineration temperatures, plastic recycling is encouraged, because it removes excess energy. Metal is confirmed as an overall priority in material recycling regardless of the moisture content of the incoming waste. PMID- 16259432 TI - Nitrogen oxides emission control options for coal-fired electric utility boilers. AB - Recent regulations have required reductions in emissions of nitrogen oxides (NOx) from electric utility boilers. To comply with these regulatory requirements, it is increasingly important to implement state-of-the-art NOx control technologies on coal-fired utility boilers. This paper reviews NOx control options for these boilers. It discusses the established commercial primary and secondary control technologies and examines what is being done to use them more effectively. Furthermore, the paper discusses recent developments in NOx controls. The popular primary control technologies in use in the United States are low-NOx burners and overfire air. Data reflect that average NOx reductions for specific primary controls have ranged from 35% to 63% from 1995 emissions levels. The secondary NOx control technologies applied on U.S. coal-fired utility boilers include reburning, selective noncatalytic reduction (SNCR), and selective catalytic reduction (SCR). Thirty-six U.S. coal-fired utility boilers have installed SNCR, and reported NOx reductions achieved at these applications ranged from 15% to 66%. Recently, SCR has been installed at >150 U.S. coal-fired utility boilers. Data on the performance of 20 SCR systems operating in the United States with low NOx emissions reflect that in 2003, these units achieved NOx emission rates between 0.04 and 0.07 lb/10(6) Btu. PMID- 16259434 TI - [Changes in the digestive apparatus in patients with the metabolic syndrome]. PMID- 16259433 TI - Application of satellite remote-sensing data for source analysis of fine particulate matter transport events. AB - Satellite sensors have provided new datasets for monitoring regional and urban air quality. Satellite sensors provide comprehensive geospatial information on air quality with both qualitative imagery and quantitative data, such as aerosol optical depth. Yet there has been limited application of these new datasets in the study of air pollutant sources relevant to public policy. One promising approach to more directly link satellite sensor data to air quality policy is to integrate satellite sensor data with air quality parameters and models. This paper presents a visualization technique to integrate satellite sensor data, ground-based data, and back trajectory analysis relevant to a new rule concerning the transport of particulate matter across state boundaries. Overlaying satellite aerosol optical depth data and back trajectories in the days leading up to a known fine particulate matter with an aerodynamic diameter of <2.5 microm (PM2.5) event may indicate whether transport or local sources appear to be most responsible for high PM2.5 levels in a certain location at a certain time. Events in five cities in the United States are presented as case studies. This type of analysis can be used to help understand the source locations of pollutants during specific events and to support regulatory compliance decisions in cases of long distance transport. PMID- 16259435 TI - [Particular features of the concurrent course of arterial hypertension and ulcer in elderly patients]. AB - The article presents a clinical course analysis in elderly patients suffering concurrently from arterial hypertension and ulcer as well as a study of the state of the mucous coat of the stomach and duodenum at the endoscopy and morphological assessment. It gives data of the daily monitoring of arterial pressure, central hemodynamics and indices of the lipid blood composition and hemorheology. There was a comparative analysis, which established the direct dependence of the ulcerous defect area on the atherogeneity index. Data of the morphological assessment seem to be most interesting as this study revealed abnormalities in microcirculation in the mucous coat of the stomach. It should be noted that disorders in the blood supply of the mucous coat of the stomach play a considerable role in the formation of ulcers in elderly patients with long-term arterial hypertension, and semination with Helicobacter pylori is very rare. PMID- 16259436 TI - [Particular features of the treatment of ulcer recurrence in patients with myocardial infarction]. AB - 107 patients with myocardial infarction and ulcer recurrence were examined. It was revealed that the ulcerous process proceeded with focal trombohemorragic disorders of local microcirculation in the gastroduodenal mucous coat in severe cases of myocardial infarction and with tromboischemic disorders of terminal blood flow in extremely severe cases. It was accompanied by the activation of acid and pepsin production, depression of mucopolysaccharides production and hypermotor gastric dyskinesis in patients with severe coronary pathology, in patients with the extremely severe pathology--by the depression of all functions of the stomach, except for normal acid production. The administration of an antiulcerous therapy helped to reduce the period of elimination of ulcer recurrence symptoms, liquidated microcirculatory and gastric functional disorders, and promoted the healing of ulcers. The therapy included omeprazole, sucralfate, long-term form of nifedipine for patients with severe myocardial infarction, and ranitidine, sucralfate and doxazosin in common doses for 20-25 days for patients with extremely severe cases. PMID- 16259437 TI - [Clinical-functional and morphological changes in the large intestine in patients with chronic abdominal ischemia]. AB - The article presents a study of clinical variants of ischemic lesions of the large intestine (ILLI) in patients with the chronic abdominal ischemia (CAI) syndrome. There was an examination of 42 patients with putative ILLI, who had a cardiovascular pathology at the background, which led to significant hemodynamic disorders of the blood flow in unpaired visceral branches of the abdominal aorta. The authors performed a complex diagnostics of ILLI including anamnestic and clinical data, laboratory and morphological assessment of the large intestine and vessels providing its blood supply. Based on the data collected, other functional and organic diseases of the large intestine (LI) were excluded. It was revealed that the final result of ILLI differed depending on the ischemia degree--from reversible functional disorders to high-grade organic lesions of the LI. Each clinical variant of ILLI has its own clinical manifestations as well as functional, organic and morphological peculiarities of changes in the LI revealed by a laboratory and histological examination of the LI. PMID- 16259438 TI - [Impact of Bestim on the immunological status and clinical course of ulcer]. AB - Ulcer disease is accompanied by all immune responses with the development of secondary immune deficiency. This makes it necessary to include immunomodulators into the complex antiulcer treatment. The application of a new drug Bestim in the combined antiulcer treatment affects the immune state of the patients favourably. It decreases the inflammatory effect in the stomach and duodenal mucosa, it precipitate regress of clinical symptoms and scarring of the ulcer. PMID- 16259439 TI - [Role of luminescence endoscopy in the diagnostics of gastroesophageal reflux disease]. PMID- 16259440 TI - [Duodenogastric refluxes and peculiarities of the duodenal ulcer course]. PMID- 16259441 TI - [H2-blockers of histamine receptors in the therapy of early stages of gastroduodenal reflux disease]. AB - The efficacy of the administration of famotidine, 20 mg BID, at early stages of gastroduodenal reflux disease (GDRD) based on a clinical and endoscopy study and treatment of patients of 0 and 1 degrees on the modified Savary-Miller scale. PMID- 16259442 TI - [Clinical value of the correction of disorders of intestinal microcenosis in patients with acid-dependent diseases of the digestive apparatus]. PMID- 16259443 TI - [Ulcer recurrence and gastroduodenal dysbacteriosis]. AB - The studies that were conducted for 82 patients with ulcer established that 22 genera of microorganisms including H. pylori in a combination from 2 up to 8 various microbial cultures can be secured from the biopsy of the mucous coat of the pre-ulcer zone at the acute phase of recurrence. They have high enzymatic and cytotoxic activity. This draws together microbiological characteristics of the ulcer defect with parameters of the infected wound and corresponds to dysbacteriosis of the third degree. The data obtained form the basis for considering that the exacerbation of ulcer is accompanied by expressed dysbiotic shifts depending on the recurrence phase. It is necessary for taking this into account in the development of methods of adequate therapy for the disease aimed at the suppression of the excess growth of mucous microflora and restoration of normal microbiocenosis in the gastroduodenal zone. PMID- 16259444 TI - [Celiac disease: up-to-date diagnostics and therapy methods]. PMID- 16259445 TI - [Formation and development of the Surgery Department of the Central Research Institute of Gastroenterology]. PMID- 16259446 TI - [Enteral tube feeding at the early postoperative period (stomach surgery)]. AB - The article describes a comparative analysis of the efficacy of various variants of nutritive support in 115 patients with complicated gastroduodenal ulcers at the early postoperative period. 95 patients received enteral tube feeding, 50 of them receiving balanced isocaloric formulas and 45 of them receiving broths and decoctions as a nutritious substrate. The control group comprised 20 patients who received full parenteral feeding. The results were assessed on Day 5 after the beginning of feeding. Satisfactory results were received in all of the groups. The main advantages of balanced formulas are their easy dosage, smaller number of complications and lower price (as compared to parenteral feeding) as well as better (as compared to "ordinary" products) indexes of the nutritious status recovery. PMID- 16259447 TI - [Adhesion and migration functions of peripheral blood leukocytes in children with chronic gastroduodenitis]. AB - The goal of the research was to study the peculiarities of adhesion and migration characteristics of leukocytes in children with chronic gastroduodenitis as compared to patients with functional dyspepsia and healthy children. An increase in the values of spontaneous adhesion and migration was noted in lymphocytes, monocytes and neutrophils during the exacerbation period of gastroduodenitis, which correlates with clinical manifestations, data of the morphological study and intragastric pH-metry. At the same time, a reduction in the functional reserve of leukocytes and their sensibility to adhesion stimulators. The dynamics of the examined characteristics was determined after the treatment. The results obtained are the evidence of the possibility to use the parameters of adhesion and migration capabilities of leukocytes for diagnosing chronic gastroduodenitis. PMID- 16259448 TI - [Immunosuppressive therapy of inflammatory diseases of the large intestine]. AB - The treatment of ulcerous colitis and Crohn's disease is based on suppression of various components of the immunoinflammatory cascade. In addition to aminosalicylates and glucocorticoids, drugs of primary immunosuppressive action are used for this purpose, which are adopted from clinical oncology and transplantation medicine: azathioprine, 6-mercaptopurine, methotrexate, cyclosporine A, tacrolimus, mycophenolate mofetil, thalidomide. Some of them are used for remission induction in case of hormonal dependence or resistance, while others are applied for its maintenance or at various stages of the inflammatory process course. The presented review discloses problems of clinical pharmacology of immunosuppressive drugs, side effects, results of pilot and clinical studies of ulcerous colitis and Crohn's disease. All this makes it possible to determine their role in the therapy of inflammatory diseases of the bowels. PMID- 16259449 TI - [Serotonergic systems of the biliary tract, stomach and duodenum in the norm and at the experimental stomach ulcer]. AB - Stimulation of the sympathetic trunk against the background of the vagus nerve irritation was shown to cause the enhancement of the electromotor activity of the gallbladder, Oddi's sphincter, stomach and duodenum blocked by ganglionary and peripheral serotonin blockers. A lateral-medial gradient of the serotonergic innervation of the biliary tract was revealed. The gastro- and duodenoprotective action of the 5-HT 1,2-serotonin blocker was established at the experimental stomach ulcer. PMID- 16259450 TI - [Systemic mechanisms of the evacuator activity of the gastroduodenal complex]. PMID- 16259451 TI - [Analysis of duodenogastric reflux symptoms in patients from Russia]. PMID- 16259452 TI - [pH, bile, and impedance monitoring in diagnosis of duodenogastric reflux]. PMID- 16259453 TI - [Diagnosis and treatment of duodenogastric reflux in aged]. PMID- 16259454 TI - [Are the proton pump inhibitors all the same?]. PMID- 16259455 TI - [Endoscopy in duodenogastric reflux ]. PMID- 16259456 TI - [What to do with Helicobacter pylori in duodenogastric reflux and why?]. PMID- 16259457 TI - [Barrett Esophagus]. PMID- 16259458 TI - [Treatment of esophagus peptic strictures]. PMID- 16259459 TI - [Treatment of extra-esophageal manifestations of gastroesophageal reflux]. PMID- 16259460 TI - [Treatment of gastroesophageal reflux: algorithm]. PMID- 16259461 TI - [Surgical treatment of gastroesophageal reflux]. PMID- 16259462 TI - [Potential risks in gastroesophageal reflux without adequate treatment]. PMID- 16259463 TI - [Historical experience of the prophylactic medical examination of the population]. PMID- 16259464 TI - Continuing care, continuing problems. PMID- 16259465 TI - Beating the bugs. PMID- 16259466 TI - Never too late to learn. PMID- 16259467 TI - How can we, as healthcare professionals, ensure that older people are given an opportunity to discuss their own resuscitation status? What impact will this have on the service we deliver? PMID- 16259469 TI - Medication management: older people and nursing. PMID- 16259468 TI - Across cultures: implementing the Evercare model. PMID- 16259470 TI - Recognising and managing stress. PMID- 16259471 TI - Walking the walk in Wales. PMID- 16259472 TI - Soul music. PMID- 16259473 TI - Alternative healthcare use in the under-served population. AB - OBJECTIVE: To apply the Behavioral Model for Vulnerable Populations to the examination of the correlates of alternative healthcare utilization among Hispanic and African-American adults residing in public housing. DESIGN: Cross sectional survey of a community-based sample. SETTING: Urban public housing communities in the county of Los Angeles. PARTICIPANTS: A geographically defined random sample of 287 African-American and Latino heads of households from three urban public housing communities. RESULTS: The use of alternative health care was assessed with three indices reflecting how frequently respondents used alternative sources of health care: 1) to prevent sickness; 2) to treat sickness; and 3) to substitute for conventional health care. Multivariate analysis of data indicates that lower education, greater perceived racial discrimination, and poorer health status were associated with the use of alternative health care to prevent sickness. Furthermore, greater perceived racial discrimination, greater financial strain, and poorer health status were associated with the use of alternative health care to treat sickness. In addition, four variables were associated with increased frequency of alternative healthcare utilization as a substitute for conventional care, namely: 1) diminished belief that powerful individuals (such as healthcare professionals) control one's health; 2) greater perception of racial discrimination; 3) greater financial strain; and 4) reduced access to health care. CONCLUSION: Enabling characteristics helped explain the use of alternative health care to treat sickness as a substitute for conventional health care, but not to prevent sickness, in this population. Perceived racial discrimination was the strongest correlate for each type of alternative healthcare use, while health status was also a strong predictor. The use of alternative health care for prevention and for substitution should be examined separately in disadvantaged minority populations. PMID- 16259474 TI - Predictors of multivitamin supplement use among African-American female students: a prospective study utilizing the theory of planned behavior. AB - BACKGROUND: Public health officials recommend that women capable of becoming pregnant use folic acid-containing supplements (FAS) to prevent neural tube defects (NTD) in their newborn infants. However, the knowledge about NTD prevention and the prevalence of the use of FAS among women capable of becoming pregnant increased only modestly since the issuing of the recommendation in 1992. Since most commonly available multivitamin supplements (MVS) contain the recommended 400 gg of folic acid, finding out reasons why women take MVS and utilizing these factors in educational campaigns may contribute to increasing the use of FAS. METHODS: The Theory of Planned Behavior variables and the self reported use of MVS were measured by two separate surveys within one week. A preliminary open-ended questionnaire was utilized to elicit beliefs about MVS. A convenience sample of 100 African-American female college students, mean age 20.99 (SD=1.7) years, participated in this study. RESULTS: Approximately 65% of variance in behavioral intention was explained by attitude, subjective norms, and perceived behavioral control (P<.001). Subjective norms had the greatest influence (3=0.348, P<.001), followed by PBC (3=0.336, P<.001), and attitude (1=0.228, P<.038). Behavioral intention significantly predicted the use of MVS accounting for =59.2% of variance. CONCLUSION: Consistent with the results of the present study, educational campaigns that target African-American female college students to encourage the use of MVS should focus on the impact of physicians, family, and peers. PMID- 16259475 TI - Traditional and western medicine: cultural beliefs and practices of South African Indian Muslims with regard to stroke. AB - OBJECTIVES: To investigate the beliefs of caregivers and traditional healers within the South African Indian Muslim community regarding the etiology and treatment of stroke and the persons likely to be consulted in this regard. DESIGN: A descriptive case study design was employed which incorporated two groups and was located within a qualitative paradigm. SETTING: Data were collected within the homes of caregivers and the consulting rooms of traditional healers. PARTICIPANTS: Ten caregivers of persons who had sustained strokes and 10 traditional healers were interviewed. INTERVENTIONS: Individual interviews were held with participants. OUTCOME MEASURES: Responses to semi-structured interview schedules were analyzed using thematic content analysis and descriptive statistics. RESULTS: For both groups, religion and faith in God played a pertinent role in beliefs regarding etiology of illnesses such as stroke. Caregivers used a combination of traditional and Western medicine approaches. For traditional healers, treatment was based on the premise of restoring the balance between hot and cold in the body, which had been placed in disequilibrium by the stroke. Participants expressed disillusionment with referrals to Western healthcare professionals whose treatment was often regarded as culturally inappropriate. They also emphasized the integral role played by family members in the treatment of illness and disease. CONCLUSIONS: Results have implications for: culturally sensitive management of stroke patients in the South African Indian Muslim community; collaboration between Western and traditional healers; involvement of families in the remediation process; and further research. PMID- 16259476 TI - Low high-density lipoprotein cholesterol is associated with impaired endothelial function in Asian Indians. AB - This study was designed to assess the relationship between plasma lipid levels and endothelial function in Asian Indians without cardiovascular risk factors living in the United States. While traditional risk factors do not account for the increased incidence of coronary heart disease (CHD) in Asian Indians, low high-density lipoprotein (HDL) cholesterol, elevated triglycerides, elevated lipoprotein (a), and insulin resistance are consistently found in Asian Indians with CHD. Endothelial function was measured in 86 healthy Asian Indians (mean age 33 years) free of cardiac risk factors with LDL levels<160 mg/dL. Subjects were divided into two groups on the basis of HDL levels (low HDL<40 mg/dL and normal HDL-40 mg/dL). Endothelial function during reactive hyperemia was significantly impaired in Asian Indians in the low HDL group. After covariate adjustment, NTG induced brachial vasodilation was not different between patients in the two HDL groups. These data indicate that low HDL is associated with endothelial dysfunction in this population. PMID- 16259477 TI - Hemoglobin A1c levels in diagnosed and undiagnosed black, Hispanic, and white persons with diabetes: results from NHANES 1999-2000. AB - PURPOSE: Although the prevalence of diabetes among various racial/ethnic groups has been well studied, little is known about the racial/ ethnic differences in Hemoglobin A1c (HbA1c) in diagnosed and undiagnosed diabetes. HbA1c correlates with morbidity and mortality in diabetes. Knowledge of the racial/ethnic differences in HbA1c would impact screening and intervention in primary care settings. This study describes racial/ethnic differences in HbA1c among US Black, Hispanic, and White diagnosed and undiagnosed persons with diabetes. METHODS: This study included participants in the 1999-2000 National Health and Nutrition Examination Survey who were > or =20 years old with a HbA1c measurement. The association between HbA1c and race in diagnosed and undiagnosed persons with diabetes (with body mass index [BMI] and age as covariates) was determined. The distribution of HbA1c and mean HbA1c in diagnosed and undiagnosed diabetes and the rates of diagnosed and undiagnosed diabetes with their corresponding HbA1c levels are described by race/ethnicity. RESULTS: Estimated diabetes prevalence in US persons > or =20 years is 8.2%, with 2.3% having undiagnosed diabetes. Whites with diabetes had lower mean HbA1c levels (7.6%, standard error [SEI 0.2) than Blacks (8.1%, SE 0.3) or Hispanics (8.2%, SE .3). Whites with diagnosed diabetes were less likely to have HbA1c> or =11% (1.7%) than Blacks (11.1%) or Hispanics (10.4%). Hispanics with undiagnosed diabetes were more likely to have HbA1c-7% (60.5%) than Blacks (39.3%) or Whites (37.8%). CONCLUSIONS: Significant numbers of persons with diabetes are undiagnosed. There are significant racial/ethnic differences in HbA1c levels, which are significantly higher in Blacks and Hispanics. Comprehensive risk-based screening and intervention for diabetes is needed in order to address racial and ethnic disparities, especially in minorities. PMID- 16259479 TI - Standard electrocardiographic criteria for left ventricular hypertrophy in Nigerian hypertensives. AB - OBJECTIVES: Left ventricular hypertrophy (LVH) is a major risk factor for cardiovascular morbidity and mortality. Various electrocardiographic criteria for LVH have differing sensitivities and specificities. Most of the available electrocardiographic criteria for LVH have not been evaluated in the African populace. METHODS: Electrocardiograms (ECGs) and echocardiograms were obtained from 100 hypertensive subjects and 60 controls. Electrocardiogram (ECG) LVH was determined by the Sokolow-Lyon, Sokolow-Lyon-Rappaport, Cornell voltage, Romhilt Estes point score, and the Perugia score criteria. Echocardiographic LVH was defined by LV mass indexed for height at 97.5 percentile of the controls (126 g/m and 130 g/m in females and males respectively). RESULTS: The prevalence of echocardiographic LVH indexed for height was 34% and 1.67% in the hypertensive patients and controls respectively. The prevalence of ECG LVH obtained in the hypertensive patients with the various ECG criteria were 56% for Sokolow-Lyon Rappaport voltage, 48% for Sokolow-Lyon voltage, 41% for Perugia score, 22% for Cornell sex specific voltage, and 18% for Romhilt-Estes score. Sokolow-Lyon Rappaport voltage criteria had the best sensitivity (80%) and area under the receiver operating characteristic (ROC) curve while the Romhilt-Estes score had the best specificity (93%). CONCLUSION: Sokolow-Lyon and Sokolow-Lyon-Rappaport voltage criteria combine the best sensitivity and specificity values and would seem better suited for the diagnosis of ECG LVH in Nigerians. PMID- 16259478 TI - Genetic and environmental determinants of lipid profile in black and white youth: a study of four candidate genes. AB - OBJECTIVE: To identify genotypes and gene-environment interactions, which may explain ethnic differences on lipid profile in Black and White youth. DESIGN, SETTING, PARTICIPANTS: Healthy adolescents and young adults (N=413, 18.6 +/-2.8 yrs, 44% Black, 53% Male) drawn from a cardiovascular study. MAIN OUTCOME MEASURES: Total cholesterol (TC), high-density lipoprotein cholesterol (HDLC), and triglyceride (TG) concentrations were obtained from frozen plasma. The ApoB Glu4154Lys, LDL receptor (LDLR) T1773C, PPARgamma Pro12Ala, and TNFalpha -308G/A polymorphisms were genotyped. Analyses adjusted for age, sex, ethnicity, body mass index (BMI), socioeconomic status (SES), and interactions. RESULTS: The ApoB Glu4154Lys polymorphism interacted with obesity and age to predict TC levels. As BMI increased, 4154Lys ApoB allele carriers had higher TC levels than 4154Glu homozygotes (difference=0.23 mmol/L at BMI=30 kg/m2, 0.54 at BMI=40, P<.05). Juvenile, but not adult, ApoB 4154Lys allele carriers had higher TC (0.34 mmol/L, P<.01). Male -308A TNFalpha allele carriers had lower HDLC (0.10 mmol/L, P<.01). Carriers of the T1 773 LDLR allele had higher TG (0.26 mmol/ L, P<.01). No effect of the PPARgamma Pro12Ala polymorphism was found; the 12Ala PPARgamma allele was rare among Blacks (2%). CONCLUSIONS: The ApoB, TNFalpha, and LDLR candidate genes influenced lipid profiles in youth independent of environmental factors. The T1773 LDLR allele, which is rare among Blacks (7%), may contribute to lower TG in Blacks. The -308A TNFalpha allele may contribute to lower HDLC in males. These gene effects and gene-environment interactions may inform prevention and treatment of atherosclerosis. PMID- 16259480 TI - Stress-related racial discrimination and hypertension likelihood in a population based sample of African Americans: the Metro Atlanta Heart Disease Study. AB - OBJECTIVE: Exposure to racial discrimination has been identified as an adverse biopsychosocial stressor that may be related to the prevalence of hypertension in African Americans. The overall objective of this research was to estimate aspects of the effects of self-reported exposure to stress-related racial discrimination and hypertension likelihood in a sample of African Americans. DATA: Physiologic and self-reported cross-sectional data were collected on a stratified sample of hypertensive (n=174) and normotensive (n=182) African Americans, aged > or =21 years, residing in metropolitan Atlanta, Georgia from 1999 through 2001. METHOD: Separate, multivariate logistic regression models were fit, including relevant covariates, to ascertain the effects of exposure to incidents of stress-related racial discrimination and level of generally derived stress with hypertension likelihood. RESULTS: Exposure to incidents of racial discrimination was not significantly associated with an increased likelihood of hypertension. More than 74% of respective normotensive and hypertensive respondents reportedly experienced stress-provoking encounters of racial discrimination. Magnitude of stress generally derived from exposure was, however, a highly significant predictor. Respondents reporting "moderate" and a "high to very high" level of derived stress were more than twice as likely to be hypertensive when compared to those reporting "no to low" derived stress (P value=.02 and .01, respectively). CONCLUSION: Exposure to racial discrimination is a prevalent psychosocial stressor in African Americans but may not be significantly associated with hypertension prevalence; degree of stress derived from encounters may be an important determinant. More research is needed to clarify the complex relationship between stress-related racial discrimination and hypertension in African Americans. PMID- 16259481 TI - The development of a race and gender-specific stress measure for African-American women: Jackson, Hogue, Phillips contextualized stress measure. AB - OBJECTIVE: Community-based research was conducted to develop an identity stress measure for African-American women. The aim of the investigation was to capture the voices of African-American women telling their experiences of stress and support and to have their voices inform the development of an identity stress measure representing the realities of being Black and female. In this paper, we describe the components of a race and gender-specific stress measure emerging from a multidisciplinary iterative process that employed qualitative and quantitative methods. METHOD: The research was initiated by focus groups and interviews where women were asked to share their experiences of stress and support. Four hundred seventy-four (474) African-American women from the metropolitan Atlanta area collaborated in the study by participating in one or more phases of the research. Content analysis of the qualitative data informed the development of a 71-item race and gender-specific stress measure for African American women. The scale and a battery of validity measures (Spielburger Anger and Anxiety, John Henryism, and NHIS-depression) were administered twice over a 30-day period followed by group discussions and interviews. RESULTS: Content and factor analysis resulted in the development of six subscales: racism, burden, personal history, work, support/coping, and stress states. The measure has been validated with established measures of anger, anxiety, depression. Significant correlations were established for all of the stress subscales and measures of anger (trait anger, anger-in, anger-out, and anger expression). Findings indicate significant correlations for the burden subscale and anger-in (r=.33, <.01) and stress states and trait anxiety (r=.57, <.01). PMID- 16259482 TI - Race/ethnic and sex differentials in pulse pressure among us adults. AB - The prevalence of high blood pressure in the United States is a public health concern. This study uses the Third National Health and Nutrition Examination Survey (1988-1994) and linear regression to document variations in pulse pressure by race/ethnicity and sex in the United States. We find higher pulse pressures among racial and ethnic minorities than among non-Hispanic Whites and among males than females. The results indicate that the effect of race on pulse pressure decreases with the inclusion of various controls; nevertheless, African Americans maintain higher pulse pressures than non-Hispanic White Americans, even net of controls. Compared to females, males exhibit higher pulse pressures. Moreover, this sex gap progressively increases with controls for socioeconomic status and physical activity. Given the known health consequences associated with high pulse pressure, these results highlight the importance of better understanding and addressing the risk of high pulse pressure among demographic subpopulations in the United States. PMID- 16259483 TI - Coronary heart disease risk factors among Cuban Americans. AB - Cuban Americans, one of three major Hispanic subgroups, represent = 4% of total US Hispanics. Coronary heart disease (CHD) is the leading cause of death among Cuban Americans. Yet, we know very little about the risk factors of CHD. Findings from one Hispanic subgroup cannot be applicable or extrapolative to other Hispanic subgroups because each subgroup's social histories, cultural identities, health behaviors, and genetic compositions are unique. This paper reviews the existing information on Cuban-American health behavior in relation to CHD. Analyses of the Hispanic Health and Nutrition Examination Survey (HHANES) data revealed a high prevalence of overweight, cigarette smoking, and type 2 diabetes among Cuban Americans. However, the cross-sectional nature of HHANES precludes identifying a cause-and-effect relationship. Well-designed studies are warranted to identify the lifestyle, biochemical, and emerging risk factors of CHD among Cuban Americans. PMID- 16259484 TI - Clinical characteristics of South Asian patients hospitalized with heart failure. AB - Ethnic variations in prevalence, presentation characteristics, and mortality have been identified in Canadian patients with coronary artery disease. Similar data with respect to heart failure do not exist. A retrospective sequential chart review of South Asians and non-South Asian Whites in Canada hospitalized with a primary diagnosis of congestive heart failure between 1997 and 1999 showed South Asians were significantly younger, of lower body mass index, were more often diabetic, and were less often smokers. In-hospital mortality was not different between groups, although South Asians were more likely to experience ventricular arrhythmias. Despite presenting at a younger age, South Asians had more high-risk features at hospital discharge. Since South Asians are at high risk of developing premature coronary artery disease, a more aggressive approach to prevention strategies in this ethnic group may reduce the subsequent burden of heart failure. PMID- 16259485 TI - Racial differences in the use of aspirin: an important tool for preventing heart disease and stroke. AB - BACKGROUND: Regular aspirin use, particularly as secondary prevention, reduces morbidity from heart disease and stroke. Few studies have examined racial/ethnic differences in aspirin use for the prevention of cardiovascular disease (CVD). METHODS: Data from the 2001 Behavioral Risk Factor Surveillance System (n=2,514 African Americans; n=865 Hispanics; n=28,038 Whites) were used to assess racial/ethnic differences in aspirin use. Multivariable logistic regression was used to examine whether the likelihood of aspirin use differs by race/ethnicity after accounting for sociodemographic and CVD risk factors. RESULTS: Regular aspirin use was associated with increasing age, male gender, lower educational attainment, hypertension, diabetes, overweight, and histories of myocardial infarction, coronary heart disease, and stroke. Aspirin use was lower among African Americans and Hispanics than Whites (28.6% and 28.7% vs 37.1%, respectively). After adjustment for sociodemographic and CVD risk factors, African Americans and Hispanics were 30%-40% less likely than Whites (OR=0.6, 95% C1=0.5, 0.7, African Americans; OR=0.7, 95% CI=0.5, 1.0, Hispanics) to take aspirin. Although aspirin use was higher among those with CVD (73.6% African Americans, 73.6% Hispanics, and 82.7% Whites), after multivariable adjustment, African Americans and Hispanics with CVD remained 30% to 50% less likely to use aspirin than Whites (OR=0.7, 95% CI=0.4, 1.2, African Americans; OR=0.5, 95% CI=0.2, 1.1, Hispanics). CONCLUSIONS: African Americans and Hispanics are less likely to take aspirin than their White counterparts. Differences in sociodemographic characteristics and CVD risk factors do not account for lower aspirin use among racial/ethnic minorities. Additional studies should examine methods to increase aspirin use in these populations. PMID- 16259486 TI - Leisure time physical activity and metabolic syndrome in Asian Indian immigrants residing in northern California. AB - OBJECTIVE: Immigrant Asian Indians possess major lipid and non-lipid risk factors that constitute features characteristic of metabolic syndrome. First-line therapy recognized in risk management of this syndrome is weight reduction and increased physical activity. We investigated the relationship of intensity and duration of leisure time physical activity (LTPA) to physiological indices of metabolic syndrome in Asian Indian immigrants. METHODS: Fifty-six apparently healthy men (43.7 years +/- 7.1; body mass index [BMI] 21-34) and women (43.1 years +/- 6.9; BMI 21-36) were screened to participate in this cross-sectional study. Leisure time physical activity (LTPA) was determined by Minnesota LTPA questionnaire. Anthropometric measurements were recorded by using standard procedures. Blood samples taken after an overnight fast were analyzed for measures defined by the NCEP ATP III criteria for metabolic syndrome. RESULTS: Prevalence of metabolic syndrome was 33.9% (age 29-59 years; average BMI 26.1 +/- 3.7) suggesting development of syndrome at younger age. While participants reported little LTPA, men were more active than women (total activity metabolic index (AMI) per week: 533 vs 204, respectively). In men, moderate activity was associated with a lower prevalence of metabolic syndrome, lower fasting glucose (r=-0.44), 2-hour glucose tolerance (r=-0.40), and lower serum triglyceride (r=-0.63). Only heavy activity was inversely associated with waist girth for both men (r=-0.46) and women (r= 0.41). Leisure activity levels reported by women were not significantly associated with any other risk factors. Low levels of physical activity were associated with prevalence of low high-density lipoprotein cholesterol (HDL-C), but reported LTPA levels were not significantly associated with favorable changes in serum HDL-C or blood pressure in both sexes. CONCLUSION: We provide evidence that Asian Indians who are physically active have a more favorable metabolic syndrome risk factor profile. Results highlight need to encourage physical activity in Asian Indian immigrants, particularly women, to reduce prevalence of metabolic syndrome. PMID- 16259487 TI - Left ventricular function in type 2 diabetes patients without cardiac symptoms in Zaria, Nigeria. AB - Fifty type 2 diabetes patients (25 of them being hypertensive) who had no cardiac symptoms had their left ventricular function assessed. There were 24 female and 26 male diabetes patients evaluated, along with a control group of 50 healthy subjects. The patients and controls underwent full clinical evaluation, which included physical examination, blood biochemistry (urea and electrolyte; creatinine, creatinine clearance; fasting blood and two-hour postprandial glucose levels, lipid profile), electrocardiograph, chest radiograph, and echocardiograph. The hypertensive diabetes patients had higher cholesterol levels, and 50% had levels >5.0 mmol/L. Sixteen patients had cataracts, 14 had background retinopathy, 12 had peripheral neuropathy, and 7 had peripheral vascular disease. The subjects had significantly lower ejection fraction than controls, and fractional shortening showed a similar pattern. Eight patients had ejection fraction <50% compared to none of the controls. Sixty-six percent of the subjects and 30% of the controls had diastolic dysfunction (reverse E/A ratio, prolonged deceleration time, and lower deceleration rate), respectively, but the diabetes patients did not show any difference. Diastolic dysfunction correlated significantly with age, fasting blood glucose, and two-hour postprandial glucose. The subjects had higher left ventricular mass (LVM) than controls. The LVM correlated significantly positively with diastolic blood pressure, systolic blood pressure, and pulse pressure. Subclinical diabetic cardiomyopathy exists in our patients; in addition, other risk factors for cardiomyopathy and coronary artery disease exist, including hypertension, hypercholesterolemia, and obesity. PMID- 16259488 TI - Discrepancies in the regulation of plasma adiponectin and TNF-alpha levels and adipose tissue gene expression in obese African Americans with glucose intolerance: a pilot study using rosiglitazone. AB - OBJECTIVES: We examined the effects of rosiglitazone: 1) on glucose homeostasis, insulin action, beta-cell function, and plasma adiponectin and TNF-alpha (TNF alpha) levels; and 2) the expression of adipose tissue TNF-alpha and adiponectin mRNA in African Americans with parental history of type 2 diabetes and with varying degrees of glucose intolerance. SUBJECTS AND METHODS: The study groups comprised 11 African Americans with normal glucose tolerance and six with diabetes and impaired glucose tolerance. The glucose-intolerant subjects received rosiglitazone (4-8 mg/day) every morning for 12 weeks. They underwent oral glucose tolerance test (OGTT) and subcutaneous adipose tissue biopsy (under local anesthesia) before and after 12 weeks of rosiglitazone therapy. Beta cell function and insulin resistance were calculated by using homeostasis model assessment (HOMA). Adipose tissue gene expression (mRNA) was measured by real time polymerase chain reaction in both groups. RESULTS: Rosiglitazone monotherapy improved both fasting and two-hour serum glucose levels during OGTT in the glucose-intolerant group. However, mean serum insulin and C-peptide levels did not change when compared with baseline. Rosiglitazone monotherapy improved insulin resistance but not overall beta-cell secretion. Mean adiponectin levels at fasting and two hours after oral glucose ingestion were significantly (50%) lower in the glucose-intolerant group than in the control group. Rosiglitazone monotherapy significantly increased plasma adiponectin levels at fasting and two hours after oral challenge by two-fold in the glucose-intolerant group. Mean plasma TNF-alpha levels were not significantly different at fasting and after two hours during OGTT. Rosiglitazone had no significant effect on plasma TNF-alpha levels during OGTT. No significant differences were seen in the expression of adipose tissue TNF-alpha and adiponectin mRNA in the groups at baseline. Rosiglitazone did not significantly change the adipose tissue adiponectin and TNF alpha mRNA. Rosiglitazone was well tolerated, without experiencing weight gain, edema, and liver function test abnormality in the glucose intolerant subjects. SUMMARY: Rosiglitazone improved glucose homeostasis and insulin resistance in high-risk African Americans. We found that adiponectin was lower in the glucose intolerant group, while TNF-alpha was similar. While rosiglitazone increased plasma adiponectin, it had no effect on adipose tissue adiponectin mRNA. In addition, rosiglitazone had no effect on plasma TNF-alpha and adipose tissue TNF alpha mRNA. We conclude that the metabolic effects of rosiglitazone could be mediated by adiponectin but not TNF-alpha in African Americans with glucose intolerance. Our study demonstrates that: 1) the role of adipocytokines in the etiology of type 2 diabetes in African Americans is complex; and 2) that adiponectin, but not TNF-alpha, could mediate the metabolic benefits of thiazolidinediones in African Americans with glucose intolerance. PMID- 16259490 TI - Ethnic disparities in glycemic control among rural older adults with type 2 diabetes. AB - Glycemic control is a predictor of diabetes-related morbidity and mortality. However, little is known about how well older adults in rural communities, with limited access to self-care resources and specialty care practitioners, control their diabetes. Even less is known about whether minority, older, rural adults are at increased risk for poor glycemic control. We analyzed data from a cross sectional survey of randomly selected older (> or =65 years) adults with type 2 diabetes in rural North Carolina. Participants (N=693) were men and women from three ethnic groups: African American, Native American, and White. Capillary blood samples were collected for HbA1C analysis. HbA1C levels (<7%, 7%-<8%, and > or =8%) were compared across ethnic and gender groups. Two multiple logistic regression models (model 1: personal characteristics; model 2: personal and health characteristics) were used to evaluate potential predictors of HbA1C > or =7%. Overall, 36.4% had HbA1C > or =7%. Native Americans and African-American men had the highest proportion at levels of poor glycemic control (> or =7%), and African-American women and White men had the lowest. In bivariate analysis, ethnicity, living arrangements, use of medications for diabetes, having a diabetes-related healthcare visit in the past year, and duration of diabetes were significantly associated with glycemic control. In multivariate analysis (model 1), being Native American, having low income without Medicaid, and being married were associated with poor glycemic control. Adding health characteristics (model 2), longer diabetes duration and diabetes medication therapy were significant predictors. These data indicate that older ethnic minorities in rural communities are at increased risk for diabetes complications and need diabetes management strategies to improve glycemic control. PMID- 16259491 TI - Ethnic comparisons of sarcopenia and obesity in diabetes. AB - OBJECTIVE: To examine the association between obesity and low relative skeletal muscle mass (sarcopenia) with type 2 diabetes and poor glycemic control, and to determine whether these associations varied by ethnicity. DESIGN, SETTING, PARTICIPANTS: Data from the Third National Health and Nutrition Examination Survey (NHANES III) was used (5,727 adults, 40-74 years of age: 26% Mexican Americans, 25% non-Hispanic Blacks, and 49% non-Hispanic Whites). MAIN OUTCOME MEASURES: Sarcopenia was defined as a skeletal muscle [SM(kg)/ height(m2)] <1 standard deviation below the young adult mean. Obesity was defined as waist circumference >102 cm in men and >88 cm in women. RESULTS: The prevalence of diabetes was 40% higher in Mexican Americans than in non-Hispanic Blacks or Whites (P<.05). The lowest prevalence of obesity and sarcopenia were observed in Mexican Americans (except for obesity in women). Independent of ethnicity, subjects with a high waist circumference were more likely to have diabetes and poor glycemic control (P<.05). These associations were strongest in non-Hispanic Whites. Conversely, sarcopenia was not associated with diabetes or poor glycemic control in any ethnic group. CONCLUSIONS: Although the prevalence of type 2 diabetes was highest in Mexican Americans, non-Hispanic Whites exhibited the highest prevalence of obesity and sarcopenia. The role of genetically based, ethnic differences in body composition and diabetes risk needs to be taken into account when developing clinical guidelines such as the waist circumference cutpoints used in this study. PMID- 16259489 TI - Diabetes management by residents in training in a municipal hospital primary care site (IPCAAD 2). AB - PURPOSE: Since diabetes is largely a primary care problem but we know little about management by residents in training--the primary care practitioners of the future--we examined surrogate outcomes reflective of their performance. METHODS: A seven-week observational study was conducted in a typical training site- a municipal hospital internal medicine resident "continuity" (primary care) clinic in a large, academic, university-affiliated training program. We evaluated control of glucose, blood pressure, and lipids; screening for proteinuria; and use of aspirin relative to national standards. RESULTS: Five hundred fifty-six (556) patients were 72% female and 97% African-American, with mean age 63 years, duration of diabetes 12 years, and BMI 34 kg/m2. Patients were managed largely with diet alone (22%) or oral agents alone (40%); 7% used oral agents and insulin in combination, and 30% insulin alone. Hemoglobin A1c (mean 8.2%) was above goal (<7.0%) in 61% of patients. Low density lipoprotein cholesterol (mean 128 mg/dL) was above goal (<100) in 76% of patients, but high density lipoprotein (mean 53 mg/dL) was at goal in 46%, and triglycerides (mean 138 mg/dL) were at goal in 85%. Diastolic pressure (mean 75 mm Hg) was at goal (<85) in 77% of patients, but systolic pressure (mean 143) was at goal (<130) in only 25% of patients. An average of only 53% of the patients had urine protein screening per 12 months, and use of aspirin was documented for only 39% of patients. CONCLUSIONS: Patients with type 2 diabetes in a typical internal medicine resident primary care clinic frequently do not achieve national standard of care goals. Since skills and attitudes developed in residency are likely to carry over into later practice, local diabetes educators may need to work with medical faculty to develop new interventions to improve postgraduate medical education in diabetes management. PMID- 16259492 TI - Evaluating a problem-based empowerment program for African Americans with diabetes: results of a randomized controlled trial. AB - OBJECTIVE: The objective of this study was to evaluate the impact of a problem based empowerment patient education program specifically tailored for urban African Americans with type 2 diabetes. RESEARCH DESIGN AND METHODS: The study used a randomized controlled trial (RCT) pretest/post-test design with repeated measures. Patients were randomly assigned to either a six-week intervention group or a six-week wait-listed control group. After completing the six sessions, patients were invited to participate in one of two follow-up conditions; attend a monthly support group or receive a monthly phone call from a nurse. Assessment measures included HbA1C, lipids, blood pressure, weight, self-management behavior and psychosocial adaptation. RESULTS: Both control and intervention patients showed a broad array of small-to-modest positive changes during the six-week RCT. These gains were maintained or improved upon during the one-year follow-up period. For patients in the two follow-up conditions, a positive correlation was seen between the number of follow-up contacts and their one-year HbA1C values. CONCLUSIONS: We believe that results of this study can be attributed to volunteer bias, study effects (ie, providing study data on several occasions to patients and their physicians during the one-year study period), and impact of the interventions. However, the study design does not allow us to examine the relative impact of these three factors on the patient improvements seen over the one-year study period. PMID- 16259493 TI - Knowledge of heart disease risk among spanish speakers with diabetes: the role of interpreters in the medical encounter. AB - OBJECTIVE: To investigate heart disease risk knowledge among Spanish speakers with diabetes. DESIGN: Single sample cross-sectional design. SETTING: A Spanish language diabetes health fair in an inner-city community center in Connecticut. PARTICIPANTS: Ninety-four Spanish-speaking adults participated. They were predominantly from Puerto Rico, had less than high school education, and were economically disadvantaged. Most had type 2 diabetes (96%) for an average of 10 years. Most had health insurance and a primary care provider. A sizable minority relied on ad hoc interpreters (friends or family members) during clinic visits, but most would prefer to use a professional medical interpreter. MEASURES AND RESULTS: Knowledge of risk for heart disease was measured by a Spanish version of the Heart Disease Fact Questionnaire (HDFQ). Knowledge of heart disease was low (mean score 17.5 [out of 25], SD=5.0), and lack of knowledge was found for the risks of diabetes, high-fat foods, cholesterol, physical activity, hypertension, family history of heart disease, sex, and whether one is necessarily cognizant of having heart disease. Regression analyses showed that bank account status and use of ad hoc interpreters contributed significantly to the prediction of HDFQ scores. Having a bank account and not using family or friends as interpreters in visits with the primary care provider predicted higher HDFQ scores. CONCLUSION: Heart disease risk knowledge was low in Spanish speakers with diabetes. Providing professional medical interpretation instead of relying on ad hoc interpreters is recommended for this high-risk group. PMID- 16259494 TI - Depression among Hispanic women with type 2 diabetes. AB - OBJECTIVE: Investigate psychosocial barriers to self-care and the prevalence of depression among Hispanic women with type 2 diabetes. RESEARCH AND DESIGN METHODS: One hundred twenty-six (126) Hispanic women living in Miami-Dade, South Florida were recruited through outpatient diabetes clinics, physicians' offices, or community referrals to participate in a cross-sectional survey to assess psychosocial factors that can interfere with type 2 diabetes self-care. The survey collected information on sociodemographics, performance of self-care regimens, a diabetes care profile (support, knowledge, empowerment, and attitudes), depression (Beck Depression Inventory-II), health locus of control, and perceived stress, as well as other data used to evaluate diabetes status. RESULTS: Only 4 of the 126 diabetic women interviewed reported having had depression previously assessed. Mean for depression scores was 12.76 +/- 8.71. Scores on the depression inventory indicated depression in 40.6% of subjects, with 23% categorized as mildly, 11.1% moderately, and 6.3% severely depressed. Greater self-assessed depression was associated with poorer self-rated health, understanding of diabetes, and diabetes knowledge scores. Subjects classified as severely depressed had lived with diabetes for a significantly longer mean number of years than those classified as moderately, mildly, or minimally depressed. CONCLUSIONS: Depression had not been previously assessed by any healthcare providers seen by participants or addressed in most participants in this study. While the literature abounds with findings on the depression in diabetes and potential for improved compliance when depression is treated, this concept seems to not yet be adopted into mainstream diabetes care for our Hispanic population. PMID- 16259495 TI - Prevalence of depressive symptoms in new Mexico Hispanic and non-Hispanic white elderly. AB - OBJECTIVE: To evaluate the prevalence rates, risk factors, and pattern of depressive symptoms in elderly Hispanic and non-Hispanic Whites (NHWs). METHODS: This survey was a community-based, cross-sectional survey of randomly selected Medicare recipients living in Bernalillo County (Albuquerque, New Mexico). The survey's objective was to examine the health and health-related issues of Hispanic and NHW elderly persons (> or =65 years of age). As part of the survey, participants were administered the Geriatric Depression Scale-short form (GDS). RESULTS: Complete data were available on 798 subjects with a mean age of 73.7 years, age range 65-96. The prevalence of a GDS score > or =6 was: Hispanic males, 9.5%; Hispanic females, 19.2%; NHW males, 5.4%; and NHW females, 8.7%. Hispanics (P=.001) and women (P=.003) had higher prevalence rates. Sociodemographic variables, health, social support, and the activities of daily living (ADLs) were significantly related to symptoms. English skills (P<.0001) and birthplace (P=.011) were associated with symptoms in Hispanics. Significant differences were found in the response patterns between Hispanics and NHWs. Logistic regression analyses showed that ethnic differences were largely explained by differences in the level of education and income among Hispanics and NHWs. CONCLUSION: A difference was seen in the prevalence rates of depressive symptoms between Hispanic and NHW elderly persons and between men and women. In addition to the traditional risk factors for depressive symptoms, we found that ethnic differences in prevalence rates can be largely explained by education and income differences in the two groups. PMID- 16259496 TI - Associations of stress and depressive symptoms with cancer in older Mexican Americans. AB - OBJECTIVES: Cancer outcomes vary by ethnicity and socioeconomic status, particularly among the elderly. Although not well understood, there is increasing evidence that psychosocial issues, including depression, may be particularly influential among older minority groups. The objective of this study was to explore associations of stress and depressive symptoms with prevalent cancer in older Mexican Americans. METHODS: Conditional logistic regression analyses were performed on baseline data from the Hispanic EPESE (collected in 1993, 1994) to explore associations of stress and depressive symptoms with prevalent cancer. RESULTS: Higher education, income, and acculturation were associated with cancer. In general, people with more education and greater income, men, and those evidencing greater acculturation were more likely to have cancer. People with cancer had also experienced more adverse life events: specific discrete life events, especially losses, were found to be more important than total number of life events. There was evidence of a J curve in the relationship of depressive symptoms with cancer, particularly for women. The presence of between three and seven symptoms was inversely associated with cancer for women, while clinical depression was positively associated with the disease. In addition, depression was positively associated with cancer in men who felt hopeless. CONCLUSIONS: In the fastest growing segment of those aged > or =65 in the United States, Mexican Americans, our findings suggest that some psychosocial factors are associated with cancer and have implications for treatment. Consistent with other research, factors associated with cancer generally reflect greater assimilation. Further exploration of subclinical depression in this older group is needed. PMID- 16259498 TI - Evaluation of African-American and white racial classification in a surveillance, epidemiology, and end results cancer registry. AB - OBJECTIVES: This study evaluated the validity of registry-reported race for individuals who participated in research studies conducted since 1980 through the Metropolitan Detroit Cancer Surveillance System (MDCSS), a Surveillance, Epidemiology, and End Results (SEER) Program registry. METHODS: 5329 individuals who self-identified as African American or White and were classified in the MDCSS registry as African American or White were included. Self-identified and registry reported race were compared, and associations between demographics and racial misclassification were examined. RESULTS: Most self-identified African Americans and Whites were correctly classified (sensitivity=98.5%, specificity=99.7%). Males were two times more likely to be misclassified than females [odds ratio (OR)=2.13, 95% confidence interval (CI): 1.06-4.29]. Individuals diagnosed with cancer after 1990 were two times more likely to be misclassified than those diagnosed before 1990 (OR= 2.17, 95% CI: 1.07-4.42). African Americans were four times more likely to be misclassified than Whites (OR=4.39, 95% CI: 2.24-8.60). CONCLUSIONS: Misclassification in the MDCSS registry of African Americans as Whites, and vice versa, is relatively low. Additional studies should evaluate misclassification of African Americans and Whites as other races and/or ethnicities in the SEER registry. PMID- 16259497 TI - Dietary patterns of reservation and non-reservation Native American youths. AB - OBJECTIVE: A leading cause of morbidity and mortality among Native Americans, cancer is largely preventable through lifestyle habits. Ranked high among those habits is a diet low in fat and high in fiber, fruits, and vegetables. Because Native Americans are not included in national nutritional monitoring systems, limited data exist on the cancer-related dietary habits of this population. To bridge this gap, this study measured the eating patterns of Native American youths in the northeastern United States. DESIGN: Cross-sectional. SETTING: Urban Indian centers and tribal and reservation settings located in New York, Massachusetts, Connecticut, Vermont, and Maine. PARTICIPANTS: 191 self-identified Native American parents of children between 8 and 14 years of age. MAIN OUTCOME MEASURES: Frequency of intake of foods recommended for inclusion in or omission from a diet to reduce cancer risk. RESULTS: Dietary patterns among Native American youths differed from a national US sample and varied between youths in reservation and non-reservation settings. CONCLUSIONS: A need exists for nutrition education and behavior change programs that reflect the culturally specific eating habits of the indigenous peoples of the Northeast. Ideally, developing nutrition curricula for specific communities will be done in cooperation with these communities and acknowledge barriers that may limit Native Americans' access to healthful foods. PMID- 16259499 TI - Barriers to breast abnormality follow-up: minority, low-income patients' and their providers' view. AB - Little is known about the factors associated with delayed or incomplete adherence to recommendations for follow-up when breast abnormalities are seen in minority women. This study examines barriers to follow-up in a cohort of predominantly minority women, with input from providers, using quantitative and qualitative methods. We conducted telephone interviews with 535 women and inperson, unstructured interviews with 31 providers from three medical facilities in the Los Angeles area. Most patient respondents were <50 years old (59.6%), Latina (84.2%), and unmarried (60.9%); half (49.1%) had six or fewer years of education, and most were foreign-born (83.4%). Data from patient and provider groups identified race/ethnicity, country of birth, financial issues, fear of pain, and difficulty navigating the healthcare system as barriers to follow-up, though certain provider-identified barriers did predict adherence among women. System barriers, not individual patient characteristics, were more salient factors in the follow-up of breast abnormalities. PMID- 16259500 TI - Explaining geographic variation in breast and cervical cancer incidence rates in US Hispanic women. AB - OBJECTIVE: This study examined geographic variation in incidence rates for two cancers common in US Hispanic women and considered some potential explanations, by using data from several high-quality cancer registries. METHODS: Age standardized incidence rates (ASIRs) per 100,000 Hispanic women per year were analyzed for breast and cervical cancer in the population-based cancer registries of the Surveillance, Epidemiology and End Results (SEER) Program. The percentage potentially misclassified (PPM) as Hispanic, because of the frequent absence of maiden name, was estimated. Sociodemographic characteristics of the cancers and of the Hispanic population in eight areas were compared. RESULTS: The ASIRs varied from 80.9 to 113.3 for breast and 8.2 to 19.9 for cervix cancers. The PPM was higher for breast (16%) than for cervix (7%). Differences in PPM across SEER areas did not appear consistent with geographic variation in ASIRs, while some variation in ASIRs was consistent with differences in sociodemographic characteristics. Registry data on subgroups (defined by ancestry and birthplace) were too incomplete for analyses of CONCLUSIONS: Some of the geographic variation appeared to be explained by sociodemographic factors, but improvements in cancer registries are needed to analyze ASIRs for Hispanic subgroups. PMID- 16259502 TI - African-American preference for same-race healthcare providers: the role of healthcare discrimination. AB - OBJECTIVE: To determine the extent to which African Americans prefer same-race clinicians and the extent to which: 1) knowledge of historical mistreatment; 2) perceptions of current racial inequities in medical treatment; and 3) personal experiences of discrimination are associated with preference for same-race healthcare providers among African Americans. DESIGN: Statistical analysis of a nationally representative telephone survey designed by the Henry J. Kaiser Family Foundation and conducted by Princeton Survey Research Associates (PSRA). Bivariate significance is determined by using chi-square tests of association. Multinominal logistic regression models adjust for age, gender, income, education, and self-reported health status. RESULTS: Approximately one in five African Americans states a preference for a same-race healthcare provider. Neither knowledge of historical mistreatment nor perceptions of current racial inequities in medical treatment are related to preferred race of healthcare providers. In contrast, personal experiences of discrimination in health care are associated with a preference for same-race healthcare providers. CONCLUSIONS: The results suggest that while knowledge of unfair treatment historically and perceptions of current racial inequity do not affect preferences, personal experiences of unfair treatment may have a significant effect on African-American patients' preferences regarding health care. Findings suggest that rather than focusing on how historical mistreatment and current inequities in medical treatment affect individual patients, research should focus on individual experiences. PMID- 16259503 TI - Knowledge, attitudes, beliefs, and blood pressure control in a community-based sample in Ghana. PMID- 16259501 TI - Acculturation and its association with health-risk behaviors in a rural Latina population. AB - PURPOSE: To evaluate the performance of proxy measures of acculturation and to examine the association between acculturation and selected health-risk behaviors. METHODS: Participants were 1062 Latina pregnant women who received prenatal care at clinics in San Joaquin County, California between 1999 and 2001. We used the receiver-operating characteristic (ROC) curve to characterize the sensitivity and specificity of proxy measures and regression analysis to examine health-risk behaviors. RESULTS: Using the ARSMA-II short version scale as a reference, age at immigration had the highest percentage of correctly classified individuals. Acculturation was significantly associated with a lifetime history of substance use, risky sexual behavior, low fruit consumption, and high fast-food meal consumption. CONCLUSIONS: Acculturation is an important predictor of health-risk behavior among women. Further research is needed to better understand the phenomenon and to avert associated adverse health consequences. PMID- 16259504 TI - Health problems and hospitalizations among Asian-American ethnic groups. AB - OBJECTIVES: To study health status and hospitalization risk among Asian Americans. DESIGN: Cross-sectional analyses and cohort study. SETTING: Comprehensive prepaid health care program in Northern California. PATIENTS: Adult Asian Americans (N=13,592), self-classified at health examinations as 6050 (44.5%) Chinese, 1707 (12.6%) Japanese, 4232 (31.1%) Filipinos, 714 (5.3%) South Asians, and 889 (6.5%) Other Asians. INTERVENTIONS: None except data analysis. OUTCOME MEASURES: With Chinese and Whites (n=72,019) as referents, comparison of symptom composites by logistic regression and hospitalization risk by Cox proportional hazards models. RESULTS: Compared to Chinese, Filipinos, South Asians, and other Asians more frequently reported coronary, respiratory, gastrointestinal, and emotional problems. Chinese and Japanese generally had the lowest hospitalization risk. Compared to Chinese, hospitalization risk was higher (P<.05) among Filipinos for circulatory (men or women), respiratory (men), and digestive (women) conditions (relative risks [RR] range from 1.5 to 1.7) and among South Asian men for cardiovascular conditions (RR=2.2). While Asian groups generally had similar or lower hospitalization risk than Whites, risks were higher for asthma (Filipino and South Asian men, RRs >3.0), peptic ulcer (Chinese men, Filipino men and women, other Asian women [RRs 1.9-5.6]), and coronary disease (South Asian men (RR=2.3) and Filipino women (RR=1.5). CONCLUSIONS: Variations in risk of hospitalization and frequency of reported health problems point out differences in health problems and health needs among subgroups of Asian Americans. This diversity shows the need to study Asian ethnic groups separately. PMID- 16259505 TI - Hepatitis B knowledge and testing among Vietnamese-American women. AB - The study objective was to examine factors associated with previous hepatitis B virus (HBV) testing among Vietnamese women. A population-based survey was conducted in Seattle. The survey was completed by 370 women (response rate: 82%). Sixty-eight percent of the respondents reported previous HBV testing. Only 44% recalled a physician recommendation for the test. The following factors were associated with previous testing in bivariate comparisons: knowing that HBV can be spread during childbirth, during sexual intercourse, and by sharing toothbrushes; doctor(s) had recommended testing; family member(s) and friend(s) had suggested testing; and family member(s) were chronically infected with HBV. In a multiple regression analysis, women who reported a previous physician recommendation had nearly a five times higher odds of testing than those who did not. Education about HBV transmission may stimulate Vietnamese women to seek testing. Intervention strategies that target social networks might be effective in increasing testing levels. Physicians should be educated about the importance of testing Asian immigrants for HBV. PMID- 16259506 TI - Hair-relaxer use and risk of preterm birth among African-American women. AB - PURPOSE: To assess whether hair-relaxer use is associated with an increased risk of preterm birth among African-American women. METHODS: We used data from the Black Women's Health Study. The participants were enrolled in 1995 and provided follow-up information in 1997, 1999, and 2001. Among 6130 singleton births reported by women <45 years of age during follow-up, 497 were preterm (<37 weeks gestation) because of premature rupture of membranes or spontaneous preterm labor for no known reason. In a case-control analysis, we compared the 497 preterm births with the 5633 births of longer gestation, and we used generalized estimation equation models to estimate multivariable odds ratios of preterm birth for users of hair relaxers. RESULTS: With control for confounding factors, the odds ratio for preterm birth among ever users of hair relaxers relative to never users was 1.0 (95% confidence interval, 0.6-1.8). No elevations were seen in risk for use started at a young age or for frequent use for long durations. CONCLUSIONS: These data suggest that hair-relaxer use does not play a role in the etiology of preterm birth in Black women. PMID- 16259507 TI - Strategies for recruiting African-American residents of public housing developments into a randomized controlled trial. AB - OBJECTIVES: Two community-based strategies used to implement a clinical trial within public housing developments are discussed: 1) hiring and training community outreach residents (CORE) team members to recruit and retain primarily African-American participants; and 2) conducting health fairs to recruit participants into a trial examining the effects of nicotine gum and motivational interviewing on smoking cessation rates. DESIGN: A cluster randomized, community based clinical trial. SETTING: This trial was conducted in housing developments within a metropolitan area in the Midwest. PARTICIPANTS: Over a period of 20 months, the research team recruited 813 residents, 80% of whom were African American, to attend health fairs. Of this number, 273 (33%) smokers were identified, and 173 were ultimately enrolled into the study. RESULTS: Attendance at health fairs of public housing development residents ranged from 8%-66% across the housing developments, with an average of 21%. A brief survey was conducted at the health fair to assess smoking status, fruit/vegetable consumption, and physical activity. CONCLUSIONS: A number of possible explanations for the relatively high participation rates among a community-based trial include engaging the community in the research process, offering free health screening services, building recruitment incentives for the CORE, and tailoring health education/promotion materials according to the demographic make-up of the developments. Details regarding the development of recruitment strategies that may boost recruitment rates in community-based clinical trials with predominantly ethnic minorities are provided. PMID- 16259508 TI - Revisiting the 1973 report, "Alarming increase of the cancer mortality in the US Black population (1950-1967)". AB - A 1973 review article by Henschke et al has been described as a "landmark," influencing the development of cancer surveillance by race/ethnicity in the United States. The 1973 article showed larger increases in total cancer mortality in Black than White males and larger increases for "non-Whites" than Whites for lung, prostate, pancreas, and various other cancers from 1950 to 1967. A review of data published after 1973 shows that the Black-White disparities in cancer mortality rates have generally increased. Research in the past 30 years supports Henschke et al's emphasis on racial differences in specific risk factors (including tobacco, alcohol, obesity, diet, and infectious diseases) and shows the importance of socioeconomic status in explaining Black-White differences in cancer risk and survival. Continued surveillance is needed to determine if declining cancer mortality rates in 1992-2001 for Blacks will continue. PMID- 16259509 TI - Esophageal cancer in African blacks of Kwazulu Natal, South Africa: an epidemiological brief. AB - BACKGROUND: Esophageal cancer is the most common carcinoma in Black South African men. A number of etiological factors have been associated with the high prevalence. OBJECTIVES: The present study was undertaken to address social and environmental factors associated with the cancer in the African Black population of KwaZulu-Natal. CASE-CONTROL STUDIES AND STATISTICAL ANALYSIS: The total number of cases recruited was 208. Of these, 87 were esophageal cancer patients, 61 nonesophageal or other-cancer patients, and 60 were non-cancer patients. We tested several symptoms and risk factors to find out causes of the cancer: eyes watering (symptom), smoking, effects of smoking and alcohol consumption combined, and consumption of beer fermented with infected maize. RESULTS AND CONCLUSIONS: Our study demonstrated statistically significant differences between the rates of eyes watering or smoking in the esophageal and the non-esophageal cancer patients. Further, a statistically significant difference was found in the rates of eyes watering between esophageal cancer and non-cancer patients. There was indeed a significant difference in the number of cases of esophageal cancer between patients who smoked or drank beer and those who did not. Patients who drank beer fermented from infected maize were more likely to have esophageal cancer than non-cancer patients. PMID- 16259510 TI - For the patient. Depression among Hispanic women with diabetes. PMID- 16259511 TI - For the patient. Understanding the health problems of Asian Americans. PMID- 16259512 TI - For the patient. Heart disease risk factors for Cuban Americans. PMID- 16259513 TI - For the patient. Why do some women avoid follow-up care for breast cancer? PMID- 16259514 TI - For the patient. Use of aspirin lower among ethnic populations. PMID- 16259515 TI - For the patient. Language is important to diabetes care. PMID- 16259516 TI - For the patient. Recruiting for clinical studies. PMID- 16259517 TI - For the patient. Prevent or control diabetes--it's important to your heart. PMID- 16259518 TI - [20-year experience in surgical treatment of popliteal aneurysm]. AB - We present our 20-year experience in surgical treatment of popliteal aneurysm in 67 patients. We conclude that popliteal aneurysms should be surgically treated as early as possible after its diagnosis, preferably before ischemic complications occur. PMID- 16259519 TI - [Prevention of the anastomosis dehiscence following low anterior rectal resections]. AB - Authors analyse 106 patients that were operated in their department by Dixon's method, in retrospective study. They analyse indications for operations, lesions distance in rectum, endosonographic and CT findings. Basic principles of the operation technique are adequate blood flow, sufficient colon mobilization and tightness of the anastomosis being supplemented with transanal pertubation. Out of early complications anastomosis dehiscence appeared in 6 patients (e.g. 5.6%) out of which 4 were treated conservatively and 2 were being reoperated on (by axial ileostomia and drainage). In discussion different opinions on preoperative preparation, neoadjuvant therapy, presacral drainage, transanal pertubation and other decompressive techniques are anticipated. PMID- 16259520 TI - [Massive spongioplasty and external fixation in the posttraumatic pseudoarthrosis management--a case review]. AB - The authors of this case review present three cases of posttraumatic pseudoarthroses as complications of open fractures of the forearm and the course of their treatment. External fixation with the Ilizar external fixation apparatus and bridging spongioplasty was applied in all patients. In the first presented case, the external fixation and spongioplasty was required after resection of the atrophic posttraumatic pseudoarthrosis. In the remaining two cases, furthermore, transposition of the distal fragment of the radius using distraction in order to correct its position against the distal part of the ulna, was required. The pseudoarthroses healed in all three subjects. The case review highlights the key significance of the external fixation method and the massive spongioplasty. PMID- 16259521 TI - [Anastomotic fistula. Complication caused by surgery]. AB - Chronic anastomotic fistula are an expression of a complication after anastomoses caused by surgery. The cause of this is diverse and reaches from incorrect seam technology, over factors of the comorbidity up to the fateful process. All regions from the esophagus to the rectum and, in addition, the biliary and pancreatic ducts can be affected. To evaluate the general problem localization diagnostics are necessary, whereby different endoscopic, radiological and combining techniques are being applied. The treatment of fistulas contains therapies of conservative, conservatively medicamentous over the intervention in endoscopic management up to the surgical treatment. In most cases the treatment of anastomotic fistula require an individual patient-fair therapy; exceptions are the pancreatic fistula. PMID- 16259523 TI - ACE inhibitors in heart failure: what more do we need to know? AB - ACE inhibitors have significantly decreased cardiovascular mortality, myocardial infarction (MI), and hospitalizations for heart failure (HF) in patients with asymptomatic or symptomatic left ventricular (LV) systolic dysfunction. Furthermore, the extended 12-year study of the SOLVD (Studies Of Left Ventricular Dysfunction) Prevention and Treatment trials (X-SOLVD) demonstrated a significant benefit with a reduction of cumulative all-cause death compared with placebo (50.9% vs 56.4%) [hazard ratio (HR) 0.86; 95% CI 0.79, 0.93; p < 0.001]. The survival benefits and significant reductions in cardiovascular morbidity related to treatment with ACE inhibitors are likely achieved by titrating the dose of ACE inhibitors to the target dose achieved in clinical trials. Although the ATLAS (Assessment of Treatment with Lisinopril And Survival) study, which randomly allocated HF patients to low- or high-dose lisinopril, showed no significant difference between groups for the primary outcome of all-cause mortality (HR 0.92; 95% CI 0.82, 1.03), the predetermined secondary combined outcome of all cause mortality and HF hospitalization was reduced by 15% for the patients receiving high-dose lisinopril compared with low-dose (p < 0.001) with a 24% reduction in HF hospitalization (p = 0.002). Despite the use of ACE inhibitors, blockade of the renin angiotensin aldosterone system (RAAS) remains incomplete, with evidence of continued production of angiotensin II by non-ACE-dependent pathways. The safety and potential benefits of angiotensin receptor antagonists (angiotensin receptor blockers [ARBs]) in patients with impaired systolic function have been assessed in moderate to large clinical trials. In patients with impaired LV systolic function and HF, combination therapy with ARBs with recommended HF therapy including ACE inhibitors in patients who remain symptomatic may be considered for its morbidity benefit. Based on the CHARM (Candesartan in Heart failure: Assessment of Reduction in Mortality and morbidity)-Added data, candesartan cilexetil in addition to standard HF therapy results in a further reduction of cardiovascular mortality. Close monitoring of renal function and serum potassium levels is needed in this setting. The VALIANT (VALsartan In Acute myocardial iNfarction Trial) results suggest that valsartan is as effective as captopril in patients following an acute MI with HF and/or LV systolic dysfunction and may be used as an alternative treatment in ACE inhibitor intolerant patients. There was no survival benefit with valsartan-captopril combination compared with captopril alone in this trial. Despite these results, ACE inhibitors remain the first-choice therapeutic agent in post-MI patients, and ARBs can be used in patients with clear intolerance. Although the use of ACE inhibitors may be appealing in patients with HF and preserved LV systolic function, there is currently no evidence from large clinical trials to support this. PMID- 16259524 TI - Pharmacological management of atrial fibrillation following cardiac surgery. AB - Atrial fibrillation (AF) is the most common complication following coronary artery bypass graft surgery (CABG). Post-CABG AF occurs most commonly on the second postoperative day and declines in incidence thereafter. A number of risk factors have been found to be associated with a higher frequency of post-CABG AF. These risk factors include advanced age, a prior history of AF, hypertension, and heart failure. Postoperative complications--including low cardiac output, use of an intra-aortic balloon pump, pneumonia, and prolonged mechanical ventilation- are also associated with higher rates of post-CABG AF. Post-CABG AF increases the risk of stroke, and the length and cost of hospitalization. Prophylactic administration of conventional beta-adrenoceptor antagonists (beta-blockers) or sotalol produces a consistent and significant reduction in the incidence of post CABG AF; however, results with prophylactic amiodarone or magnesium are less consistent. Termination of post-CABG AF, once it occurs, can be accomplished with a number of antiarrhythmic agents. Ibutilide has been the most widely studied agent for this indication. Sotalol is not indicated for cardioversion of AF and has not been studied in the post-CABG setting. Electrical cardioversion and biatrial pacing have also been used to terminate post-CABG AF. Ventricular rate is best controlled with beta-blockers and calcium channel antagonists. Esmolol has a rapid onset of action and is easily titrated to effect. Digoxin can control the ventricular rate, but has a slow onset of action. There are limited data available to guide decisions regarding the optimal management of post-CABG AF. PMID- 16259525 TI - Antiarrhythmic drugs in patients with implantable cardioverter-defibrillators. AB - Antiarrhythmic drugs need to be initiated in up to 70% of patients with implantable cardioverter-defibrillators (ICDs) in order to treat atrial tachyarrhythmias, decrease the frequency of defibrillator shocks, and terminate ventricular arrhythmias along with antitachycardia pacing. trial fibrillation (AF) occurs in about 20% of patients with ICDs (the majority with congestive heart failure [CHF]). Antiarrhythmic drugs are initiated for this indication in 2 20% of the ICD population. Data from CHF-STAT (Congestive Heart Failure: Survival Trial of Antiarrhythmic Therapy; amiodarone vs placebo) and DIAMOND-AF (Danish Investigations of Arrhythmia and Mortality ON Dofetilide--rial Fibrillation; dofetilide vs placebo) support the approach that restoration and maintenance of sinus rhythm might be beneficial in CHF, even though no study has specifically addressed the CHF population with ICDs. Further clarification on potential benefits of rhythm control in CHF-associated AF will come from the AF-CHF (Atrial Fibrillation and Congestive Heart Failure) trial that is currently underway. The vast majority of patients with ICDs will have discharges of their devices during follow-up. Although class III antiarrhythmic drugs are widely considered to be effective for prophylaxis against frequent shocks, there are surprisingly few controlled studies that demonstrate this. In contrast to conflicting amiodarone data, sotalol has been found to be effective in preventing shocks from ICDs in prospective, randomized, placebo-controlled studies. A large study (SHIELD; SHock Inhibition Evaluation with azimiLiDe) has shown that azimilide significantly reduces ventricular tachyarrhythmia recurrence, thereby reducing the burden of symptomatic ventricular tachyarrhythmia. Other novel antiarrhythmic drugs, such as dofetilide or dronedarone, as well as different strategies (e.g. in the OPTIC [Optimal Pharmacological Therapy in Implantable Cardioverter] trial; beta adrenoceptor antagonist therapy alone, amiodarone plus beta-adrenoceptor antagonist therapy, or sotalol alone) for the prevention of ICD shocks are under evaluation. The majority of antiarrhythmic drugs, including sotalol, dofetilide, and azimilide, have no effect on, or are even associated with a decrease in, defibrillation thresholds in humans. Amiodarone, in contrast, has been shown to be related to higher defibrillation thresholds at implant and during follow-up of monophasic devices. Potential cardiac (e.g. ventricular proarrhythmia, negative inotropic effect) and drug-specific non-cardiac adverse effects are a frequent cause for drug discontinuation and need to be considered when initiating and maintaining antiarrhythmic drug therapy. In conclusion, antiarrhythmic drugs are frequently used in ICD patients, the main indications being treatment of atrial tachyarrhythmias and prevention of ICD shocks. Despite potential adverse effects, antiarrhythmics can be administered safely, as long as ICD/drug interactions are appreciated. Controlled studies that will further define the role of concomitant antiarrhythmic drug utilization in patients with ICDs are underway. PMID- 16259526 TI - Beyond low-density lipoprotein: addressing the atherogenic lipid triad in type 2 diabetes mellitus and the metabolic syndrome. AB - The metabolic syndrome and type 2 diabetes mellitus are both becoming more prevalent, and both increase the risk of cardiovascular disease. Many patients are not receiving appropriate treatment for the type of dyslipidemia that commonly occurs in these disorders--the so-called 'atherogenic lipid triad' of high serum triglyceride levels, low serum high-density lipoprotein cholesterol (HDL-C) levels, and a preponderance of small, dense, low-density lipoprotein cholesterol (LDL-C) particles. All of the processes involved in atherogenesis can be exacerbated by insulin resistance and/or the metabolic syndrome. Hypertriglyceridemia is a strong predictor of coronary heart disease. There is also an inverse relationship between serum levels of HDL-C and triglycerides in diabetic patients, with low serum HDL-C levels possibly representing an independent risk factor for cardiovascular disease. Small, dense, LDL-C particles are also highly atherogenic as they are more likely to form oxidized LDL and are less readily cleared. Insulin resistance, which is central to the metabolic syndrome and type 2 diabetes mellitus, leads to high levels of very low-density lipoprotein (VLDL), which contain a high concentration of triglycerides, resulting in high serum triglyceride levels and low serum HDL-C levels. Even though modification of the atherogenic lipid triad is probably one of the most effective methods of reducing cardiovascular risk, therapy for diabetic dyslipidemia is often directed to first lowering serum LDL-C levels with a HMG CoA reductase inhibitor. This may leave substantial excess risk for cardiovascular disease in patients with these types of dyslipidemia. The results of recent trials evaluating HMG-CoA reductase inhibitors have been mixed, with two showing no significant effect on cardiovascular outcomes in subgroups of diabetic patients. The recent CARDS (Collaborative Atorvastatin Diabetes Study) showed that atorvastatin can reduce cardiovascular events in a trial specifically designed for a diabetic population, though the population had to have at least one other risk factor in addition to diabetes mellitus. Fibric acid derivatives, such as fenofibrate, bezafibrate and gemfibrozil, are potentially well suited to the treatment of dyslipidemia that is generally associated with type 2 diabetes mellitus and the metabolic syndrome, as they are usually more effective than HMG CoA reductase inhibitors for normalizing serum levels of HDL-C and triglycerides. Promising results have been obtained from several trials of fibric acid derivatives including the BIP (Bezafibrate Infarction Prevention) study and the VA-HIT (Veterans Affairs Cooperative Studies Program HDL-C Intervention Trial; gemfibrozil). The FIELD (Fenofibrate Intervention and Event Lowering in Diabetes) trial, a clinical outcomes trial specifically designed to evaluate fenofibrate in a large population of patients with type 2 diabetes mellitus, many of whom have the metabolic syndrome, is underway. The FIELD trial results should shed light on the efficacy and safety of fenofibrate in reducing cardiovascular morbidity in diabetic and metabolic syndrome patients and on the safety profile of combination therapy with fenofibrate and a HMG-CoA reductase inhibitor. PMID- 16259527 TI - Peroxisome proliferator-activated receptor-gamma and its agonists in hypertension and atherosclerosis : mechanisms and clinical implications. AB - Cardiovascular diseases are the leading cause of morbidity and mortality in the US. Proper management and/or prevention of atherosclerosis and hypertension, two complex and chronic disorders, would significantly reduce the risk for cardiovascular events such as myocardial infarction and stroke, but this requires an understanding of the mechanisms underlying their development and progression. Whereas a great deal has been learned and applied toward the management of these disorders, especially hypertension, morbidity and mortality remains unacceptably high, most likely because there are disease-causing mechanisms that have yet to be fully recognized. Understanding these disease mechanisms is necessary so that novel management strategies can be developed. One of these novel mechanisms centers on peroxisome proliferator-activated receptor (PPAR)-gamma. PPAR-gamma is a member of the nuclear receptor superfamily of ligand-activated transcription factors known to play a role in glucose homeostasis and adipocyte differentiation and, more recently, has been shown to have anti-inflammatory, antiatherogenic, and antihypertensive effects. Thiazolidinediones, a class of drugs used in the treatment of type 2 diabetes mellitus, are high-affinity ligands for PPAR-gamma. In this review, the anti-inflammatory, anti-atherosclerotic, and anti hypertensive mechanisms by which PPAR-gamma and its agonists are thought to exert protective effects on the cardiovascular system are discussed. Ongoing clinical trials using PPAR-gamma activators for the management of cardiovascular diseases, especially in patients with type 2 diabetes mellitus, are summarized. PMID- 16259528 TI - The role of the platelet in the pathogenesis of atherothrombosis. AB - Platelet adhesion, activation, and aggregation at sites of vascular endothelial disruption caused by atherosclerosis are key events in arterial thrombus formation. Platelet tethering and adhesion to the arterial wall, particularly under high shear forces, are achieved through multiple high-affinity interactions between platelet membrane receptors (integrins) and ligands within the exposed subendothelium, most notably collagen and von Willebrand factor (vWF). Platelet adhesion to collagen occurs both indirectly, via binding of the platelet glycoprotein (GP) Ib-V-IX receptor to circulating vWF, which binds to exposed collagen, and directly, via interaction with the platelet receptors GP VI and GP Ia/IIb. Platelet activation, initiated by exposed collagen and locally generated soluble platelet agonists (primarily thrombin, ADP, and thromboxane A2), provides the stimulus for the release of platelet-derived growth factors, adhesion molecules and coagulation factors, activation of adjacent platelets, and conformational changes in the platelet alpha(IIb)beta3 integrin (GP IIb/IIIa receptor). Platelet aggregation, mediated primarily by interaction between the activated platelet GP IIb/IIIa receptor and its ligands, fibrinogen and vWF, results in the formation of a platelet-rich thrombus. Currently available antiplatelet drugs (aspirin [acetylsalicylic acid], dipyridamole, clopidogrel, ticlopidine, abciximab, eptifibatide, tirofiban) act on specific targets to inhibit platelet activation and aggregation. Elucidation of the multiple mechanisms involved in platelet thrombus formation provides opportunities for selectively inhibiting the pathways most relevant to the pathophysiology of atherothrombosis. PMID- 16259529 TI - Pharmacological prevention of venous thromboembolism in medical patients at risk. AB - Acutely ill general medical patients are at moderate-to-high risk of venous thromboembolism (VTE); approximately 10-30% may develop deep vein thrombosis or pulmonary embolism, the latter being a leading contributor to deaths in hospital. Medical conditions associated with a high risk of VTE include cardiac disease, cancer, respiratory disease, inflammatory bowel disease, and infectious disease. Predisposing risk factors for VTE in medical patients include history of VTE, history of malignancy, complicating infections, increasing age, thrombophilia, prolonged immobility, and obesity. Unfractionated heparin (UFH), low-molecular weight heparin (LMWH), and fondaparinux sodium have been shown to be effective agents in the prevention of VTE in medical patients. In this setting, UFH has a higher rate of bleeding complications than LMWH. There is no evidence supporting the use of aspirin, warfarin, or mechanical methods to prevent VTE in medical patients. We recommend either LMWH or fondaparinux sodium as well tolerated and effective thromboprophylactic agents in medical patients. PMID- 16259530 TI - Heritability of malaria in Africa. AB - BACKGROUND: While many individual genes have been identified that confer protection against malaria, the overall impact of host genetics on malarial risk remains unknown. METHODS AND FINDINGS: We have used pedigree-based genetic variance component analysis to determine the relative contributions of genetic and other factors to the variability in incidence of malaria and other infectious diseases in two cohorts of children living on the coast of Kenya. In the first, we monitored the incidence of mild clinical malaria and other febrile diseases through active surveillance of 640 children 10 y old or younger, living in 77 different households for an average of 2.7 y. In the second, we recorded hospital admissions with malaria and other infectious diseases in a birth cohort of 2,914 children for an average of 4.1 y. Mean annual incidence rates for mild and hospital-admitted malaria were 1.6 and 0.054 episodes per person per year, respectively. Twenty-four percent and 25% of the total variation in these outcomes was explained by additively acting host genes, and household explained a further 29% and 14%, respectively. The haemoglobin S gene explained only 2% of the total variation. For nonmalarial infections, additive genetics explained 39% and 13% of the variability in fevers and hospital-admitted infections, while household explained a further 9% and 30%, respectively. CONCLUSION: Genetic and unidentified household factors each accounted for around one quarter of the total variability in malaria incidence in our study population. The genetic effect was well beyond that explained by the anticipated effects of the haemoglobinopathies alone, suggesting the existence of many protective genes, each individually resulting in small population effects. While studying these genes may well provide insights into pathogenesis and resistance in human malaria, identifying and tackling the household effects must be the more efficient route to reducing the burden of disease in malaria-endemic areas. PMID- 16259531 TI - Maternal malaria and gravidity interact to modify infant susceptibility to malaria. AB - BACKGROUND: In endemic areas, placental malaria due to Plasmodium falciparum is most frequent and severe in first-time mothers, and increases the risk of infant mortality in their offspring. Placental malaria may increase the susceptibility of infants to malaria parasitemia, but evidence for this effect is inconclusive. METHODS AND FINDINGS: During 2002-2004, we monitored parasitemia in 453 infants, including 69 who were born to mothers with placental malaria, in a region of northeastern Tanzania where malaria transmission is intense. We used a Cox proportional hazards model to evaluate the time from birth to first parasitemia, and a generalized estimating equations logistic regression model to evaluate risk of any parasitemia throughout the first year of life. Compared with infants whose mothers did not have placental malaria at delivery ("PM-negative"), offspring of mothers with placental malaria at delivery ("PM-positive") were 41% more likely to experience their first parasitemia at a younger age (adjusted hazard ratio [AHR] = 1.41, 95% confidence interval [CI] 1.01-1.99). The odds of parasitemia throughout infancy were strongly modified by the interaction between placental malaria and gravidity (p for interaction = 0.008, Type 3 likelihood ratio test). Offspring of PM-negative primigravidae had lower odds of parasitemia during infancy (adjusted odds ratio [AOR] = 0.67, 95% CI 0.50-0.91) than offspring of PM negative multigravidae, and offspring of PM-positive primigravidae had the lowest odds (AOR = 0.21, 95% CI 0.09-0.47). In contrast, offspring of PM-positive multigravidae had significantly higher odds of parasitemia (AOR = 1.59, 95% CI 1.16-2.17). CONCLUSION: Although parasitemia is more frequent in primigravid than multigravid women, the converse is true in their offspring, especially in offspring of PM-positive women. While placental malaria is known to increase mortality risk for first-born infants, it surprisingly reduced their risk of parasitemia in this study. Placental malaria of multigravidae, on the other hand, is a strong risk factor for parasitemia during infancy, and therefore preventive antimalarial chemotherapy administered to multigravid women close to term may reduce the frequency of parasitemia in their offspring. PMID- 16259532 TI - Prevalence, correlates, and comorbidity of bipolar I disorder and axis I and II disorders: results from the National Epidemiologic Survey on Alcohol and Related Conditions. AB - OBJECTIVE: To present nationally representative data on 12-month and lifetime prevalence, correlates, and comorbidity of bipolar I disorder. METHOD: The data were derived from the 2001-2002 National Epidemiologic Survey on Alcohol and Related Conditions (N = 43,093). Prevalences and associations of bipolar I disorder with sociodemographic correlates and Axis I and II disorders were determined. RESULTS: Prevalences of 12-month and lifetime DSM-IV bipolar I disorder were 2.0% (95% CI = 1.82 to 2.18) and 3.3% (95% CI = 2.76 to 3.84), respectively, and no sex differences were observed. The odds of bipolar I disorder were significantly greater among Native Americans, younger adults, and respondents who were widowed/separated/divorced and of lower socioeconomic status and significantly lower among Asians and Hispanics (p < .05). Men were significantly (p < .05) more likely to have unipolar mania and earlier onset and longer duration of manic episodes, while women were more likely to have mixed and major depressive episodes and to be treated for manic, mixed, and major depressive episodes. Bipolar I disorder was found to be highly and significantly related (p < .05) to substance use, anxiety, and personality disorders, but not to alcohol abuse. CONCLUSION: Bipolar I disorder is more prevalent in the U.S. population than previously estimated, highlighting the underestimation of the economic costs associated with this illness. Associations between bipolar I disorder and Axis I and II disorders were all significant, underscoring the need for systematic assessment of comorbidity among bipolar I patients. PMID- 16259533 TI - Aripiprazole augmentation of antidepressants for the treatment of partially responding and nonresponding patients with major depressive disorder. AB - OBJECTIVE: To determine the efficacy and tolerability of aripiprazole, a dopamine D2 and 5-HT1A receptor partial agonist, as augmentation of antidepressant treatment of partially responding and nonresponding patients with major depressive disorder. METHOD: Fifteen patients with major depressive disorder (diagnosed with a site-generated form described in the text) and an incomplete response or no response to > or = 8 weeks of antidepressant (selective serotonin reuptake inhibitor, venlafaxine, or bupropion) monotherapy were treated with aripiprazole augmentation in an 8-week, open-label study. Data were gathered from July 2003 to March 2004. RESULTS: The mean duration of antidepressant monotherapy at baseline was 43.1 weeks. At baseline, mean Clinical Global Impressions Severity of Illness scale and Hamilton Rating Scale for Depression (HAM-D) scores were 4.3 and 18.9, respectively. After initiation of aripiprazole augmentation, 6 of 15 patients achieved remission (HAM-D score < or = 7) at week 1, and 9 of 15 patients remitted by week 2. All 8 completers achieved remission by study endpoint. Akathisia in 2 patients who withdrew prematurely prompted a reduction in the starting dose of aripiprazole from 10 mg/day to 2.5 mg/day, resulting in a 50% reduction in attrition due to akathisia (2/7 withdrew due to akathisia with the 10-mg starting dose, 1/8 withdrew due to akathisia with the 2.5-mg starting dose). Discontinuation rates after 4 weeks of treatment were lower for the 2.5-mg starting dose (1/8 patients) than for the 10-mg starting dose (3/7 patients). Overall discontinuation rates at endpoint were lower for the 2.5-mg dose (3/8 patients) than the 10-mg dose (4/7 patients). Response to aripiprazole augmentation did not appear to be related to the antidepressant used at study initiation. CONCLUSION: Aripiprazole is an effective augmentation strategy for improving therapeutic response in patients with treatment-resistant major depressive disorder when administered in combination with standard antidepressant therapy. Based on this clinical signal, a double-blind, placebo-controlled trial is warranted. PMID- 16259534 TI - Prevalence and features of intermittent explosive disorder in a clinical setting. AB - OBJECTIVE: To determine the lifetime and current prevalence, along with other characteristics such as age at onset, patterns of comorbidity, and interest in treatment, of DSM-IV intermittent explosive disorder (IED) in an outpatient psychiatric sample. METHOD: 1300 individuals presenting for outpatient psychiatric treatment at Rhode Island Hospital, Providence, R.I., underwent structured diagnostic assessment for Axis I and II disorders. The diagnosis of IED was made according to DSM-IV criteria. RESULTS: 6.3% (SE, +/- 0.7%) of patients met criteria for lifetime DSM-IV IED, and 3.1% +/- 0.5% of patients met criteria for current DSM-IV IED. While DSM-IV IED was the current principal diagnosis in only 0.6% +/- 0.2% of patients, most patients with current DSM-IV IED (80%) were interested in treatment for their intermittent aggressive behavior. Only lifetime alcohol/drug disorder was more frequent in DSM-IV IED compared with non-IED patients. Age at onset for DSM-IV IED peaked in the teen years, was earlier for men than women, and occurred earlier than all comorbid disorders, with the exception of phobic anxiety disorders, suggesting that IED cannot be attributed to most comorbid conditions. CONCLUSIONS: DSM-IV IED in psychiatric samples is far more common than previously thought. DSM-IV IED develops early in life, especially in male patients, and its development may be independent of most other disorders. PMID- 16259535 TI - Paroxetine response and tolerability among ethnic minority patients with mood or anxiety disorders: a pooled analysis. AB - BACKGROUND: Because of the poor quality of mental health care received by minorities, analyses documenting comparable response to and tolerability of medications for anxiety and depression in large samples of minority and majority populations could increase the willingness of providers and patients to use medications in minority populations. METHOD: A pooled analysis of 14,875 adults who participated in 104 double-blind, placebo-controlled paroxetine clinical trials investigating major depression, panic disorder, generalized anxiety disorder, social anxiety disorder, obsessive-compulsive disorder, posttraumatic stress disorder, or premenstrual dysphoric disorder from March 1984 through March 2002. An intent-to-treat analysis with last observation carried forward used the Clinical Global Impressions (CGI) scale to measure dichotomous outcome, classified as either response (CGI score of 1 or 2) or more complete response (CGI score of 1) ("full response"). Minority group differences were examined using logistic regression for the entire sample and repeated for those with major depression. Adverse events greater than 5% and twice the rate of placebo were descriptively tabulated. Finally, a survival analysis examined group differences in speed of onset of response. RESULTS: Hispanic and Asian subjects had a slightly lower response rate, while Asians had the highest rates and Hispanics had the lowest rates of "full response." The more consistent Hispanic outcome differences appeared to be due to a higher placebo response rate. There was no treatment by minority group interaction for depressed patients. Speed of response and adverse effects were similar across groups. CONCLUSIONS: There were few consistent differences in medication response and tolerability. These findings may serve to counteract the greater rate of negative attitudes toward medication use among minorities and reinforce the value of medications used to treat anxiety and depression in minorities. PMID- 16259536 TI - Augmentation with open-label atomoxetine for partial or nonresponse to antidepressants. AB - BACKGROUND: Atomoxetine is a selective nor-epinephrine reuptake inhibitor currently approved for the treatment of attention-deficit/hyperactivity disorder. Other compounds that enhance synaptic norepinephrine have shown efficacy as antidepressant monotherapies and as augmentation agents. This case series study examined the role of atomoxetine in antidepressant augmentation. METHOD: Fifteen adult outpatients with primary DSM-IV Axis I depressive disorders received open label atomoxetine augmentation following partial response or nonresponse to at least 8 weeks of standard antidepressant pharmacotherapy. Atomoxetine 40 mg/day was added to ongoing medication regimens and titrated according to clinical response. Atomoxetine was systematically offered to patients from July through October 2003. RESULTS: Eleven patients (73%) completed at least 6 weeks of atomoxetine augmentation. Mean endpoint dose was approximately 80 mg/day. Nine patients (60%) met criteria for positive categorical response. Inventory of Depressive Symptomatology-Self-Report scores decreased significantly from baseline to endpoint, and clinician ratings of social and occupational functioning increased. There were no significant changes in heart rate or blood pressure, and the most common side effect was activation. A modest but significant drop in body mass index was observed (p = .025), and a subset (6/15; 40%) of patients reported improved sexual function. CONCLUSION: More studies are warranted to evaluate the potential utility of atomoxetine for antidepressant augmentation. PMID- 16259537 TI - Diagnostic stability 18 months after treatment initiation for first-episode psychosis. AB - OBJECTIVES: (1) Assessment of diagnostic stability of psychotic disorders or psychotic mood disorders from 6 weeks to 18 months after initiation of treatment in a representative first-episode psychosis (FEP) sample. (2) Comparison between those patients who shifted from DSM-IV schizophreniform disorder to schizophrenia or schizo-affective disorder and those whose diagnosis of schizophreniform disorder remained stable. METHOD: The Early Psychosis Prevention and Intervention Centre (EPPIC) in Australia admitted 786 FEP patients from January 1998 to December 2000. Data were collected from patients' medical records (MRs) using a standardized questionnaire. Seven hundred four MRs were available, 36 of which were excluded owing to nonpsychotic diagnoses or a psychotic disorder due to a general medical condition. Of the remaining 668 patients, 176 (26.3%) were lost to follow-up. Four hundred ninety-two subjects were analyzed. Strategies to assure validity and reliability of diagnoses were applied. RESULTS: The same diagnosis was made at baseline (< or = 6 weeks after admission into EPPIC) and 18 months for 69.9% of the patients. Among the most consistent diagnoses were schizophrenia (97.3%), schizoaffective disorder (94.1%), and bipolar disorder (83.2%); the least stable, as expected, was schizophreniform disorder (40.0%). In subjects with schizophreniform disorder at baseline, the best predictors of a shift from schizophreniform disorder to schizophrenia or schizoaffective disorder were a higher baseline Clinical Global Impressions-Severity of Illness scale score and lower premorbid Global Assessment of Functioning score, although the variance accounted for was small (R2 = .07). CONCLUSIONS: A longitudinally based diagnostic process in FEP samples is needed, especially in schizophreniform disorder and bipolar disorder. However, a thorough initial assessment of patient and family by a specialized team of investigators regarding the kind and duration of patient symptoms may lead to high diagnostic stability, especially in schizophrenia and schizoaffective disorder, even in a FEP sample with a relatively short duration of untreated psychosis. PMID- 16259538 TI - Dissociative disorders among inpatients with drug or alcohol dependency. AB - OBJECTIVE: To determine the prevalence of dissociative disorders among inpatients with alcohol or drug dependency. METHOD: The Dissociative Experiences Scale was used to screen 215 consecutive inpatients admitted to the dependency treatment center of a large mental hospital over a 1-year period (March 1, 2003, to March 31, 2004). Patients who had scores of 30.0 or above were compared with patients who scored below 10.0 on the scale. The patients in both groups were then evaluated using the Dissociative Disorders Interview Schedule and the Structured Clinical Interview for DSM-IV Dissociative Disorders. The interviewers were blind to the Dissociative Experiences Scale scores. RESULTS: Of the patients, 36.7% had a Dissociative Experiences Scale score of 30.0 or above. The prevalence of DSM-IV dissociative disorders was 17.2% (N = 37). On average, 64.9% of these patients' dissociative experiences had started 3.6 years (SD = 2.9; range, 1.0-11.0 years) before onset of the substance use. Patients with dissociative disorders were younger, and the mean duration of their remission periods was shorter. Dissociative disorder patients tended to use more than 1 substance, and drugs were used more frequently than alcohol in this group. The frequency of borderline personality disorder, somatization disorder, history of suicide attempt, and childhood abuse and neglect occurred more frequently in the dissociative disorder group than in the nondissociative disorder group. History of suicide attempt (p = .005), female sex (p = .050), and childhood emotional abuse (p = .010) were significant predictors of a dissociative disorder diagnosis. Significantly more patients with dissociative disorders stopped their treatment prematurely (p < .001). CONCLUSION: Impact of dissociative disorders on development and treatment of substance dependency requires further study. PMID- 16259539 TI - Sleep and depression. AB - BACKGROUND: Of all the psychiatric disorders associated with insomnia, depression is the most common. It has been estimated that 90% of patients with depression complain about sleep quality. Since the first reports of short rapid eye movement (REM) latency in depressed patients and of the effect of sleep deprivation on depression in the 1970s, numerous sleep studies have provided extensive observations and theoretical hypotheses concerning the etiology and pathophysiology of depression. The aim of this review is to summarize knowledge regarding the relationships between sleep and depression. DATA SOURCES AND SELECTION: MEDLINE and PsycINFO searches of the literature published in English or French between 1964 and 2005 that examined the relationships between sleep disturbance and depression were conducted. Search terms used were depression, depressive disorder, affective disorder, mood disorders, seasonal affective disorder, sleep, sleep disorders, insomnia, REM, polysomnography, sleep deprivation, electroencephalography, PET, SPECT, and fMRI. DATA SYNTHESIS: Two hundred five papers were identified and selected and then integrated into the following categories: sleep architecture, antidepressive therapies, age- and gender-associated differences, functional imaging results, and sleep-related hypotheses explaining the pathophysiology of depression. CONCLUSION: Numerous studies provide findings indicating the remarkable relationship between sleep alterations and depression. Although the existing hypotheses are not likely to explain all aspects of the sleep alterations in depression, each may be worth being maintained for refinements of pathophysiologic models of depression as new data accumulate. Further research taking into account the heterogeneity of depressive disorder and linking the different areas of research is needed to develop more comprehensive theoretical models and new therapies for depression. PMID- 16259540 TI - A 24-week randomized, double-blind, placebo-controlled study of escitalopram for the prevention of generalized social anxiety disorder. AB - OBJECTIVE: Escitalopram has proven efficacy in the short-term treatment of generalized social anxiety disorder (SAD). The present relapse prevention study investigated relapse rates during a 24-week, randomized, double-blind, placebo controlled period in patients with generalized SAD who had responded to 12-week open-label treatment with escitalopram. METHOD: A total of 517 patients with a primary diagnosis of generalized SAD (per DSM-IV criteria) and a Liebowitz Social Anxiety Scale (LSAS) total score of > or = 70 received 12 weeks of open-label treatment with flexible doses (10-20 mg/day) of escitalopram. Of these patients, 371 responded (Clinical Global Impressions-Improvement scale [CGI-I] score of 1 or 2) and were randomly assigned to 24 weeks of double-blind treatment with escitalo-pram (10 or 20 mg/day) (N = 190) or placebo (N = 181), continuing with the dose level administered at the end of the open-label period. Relapse was defined as either an increase in LSAS total score of > or = 10 or withdrawal due to lack of efficacy, as judged by the investigator. The study was conducted from January 2001 to June 2002. RESULTS: Survival analysis of relapse and time to relapse showed a significant advantage for escitalopram compared to placebo (log rank test: p < .001). The risk of relapse was 2.8 times higher for placebo treated patients than for escitalopram-treated patients (p < .001), resulting in significantly fewer escitalopram-treated patients relapsing (22% vs. 50%), at both doses. Escitalopram was well tolerated during double-blind treatment of generalized SAD, and only 2.6% of the escitalopram-treated patients withdrew because of adverse events. The overall discontinuation rate, excluding relapses, was 13.2% for patients treated with escitalopram and 8.3% for patients treated with placebo. CONCLUSION: Escitalopram was effective and well tolerated in the long-term treatment of generalized SAD. PMID- 16259541 TI - Associations of anxiety-related symptoms with reported history of childhood sexual abuse in schizophrenia spectrum disorders. AB - OBJECTIVE: Research suggests that persons with schizophrenia tend to experience significant levels of anxiety and that history of childhood sexual abuse may predispose some with schizophrenia to experience significant levels of persistent anxiety. It is unclear whether childhood sexual abuse is more closely linked to specific forms of anxiety including symptoms of post-traumatic stress disorder (PTSD). METHOD: Data were gathered from April 2004 through November 2004 on trauma history, PTSD symptoms, social anxiety, and state and trait anxiety from 45 men with a SCID-I-confirmed diagnosis of schizophrenia or schizoaffective disorder and 11 with a SCID-I-confirmed diagnosis of PTSD with no history of psychosis. Participants with schizophrenia spectrum disorders (schizophrenia group) were divided into those with and without history of childhood sexual abuse. Five participants in the schizophrenia group with a history of adult but not childhood sexual assault were excluded from analyses. RESULTS: Analysis of variance comparing the childhood sexual abuse (N = 21) and non-abused (N = 19) schizophrenia groups and the PTSD group on all anxiety assessments revealed that the sexually abused schizophrenia group had significantly higher levels of dissociation, intrusive experiences, and state and trait anxiety than the non abused schizophrenia group. The schizophrenia groups did not differ statistically on levels of anxious arousal, defensive avoidance, or social anxiety. When compared with participants with PTSD and no psychosis, the sexually abused schizophrenia group had significantly lower levels of state anxiety, anxious arousal, intrusive experiences, and fearful social avoidance but failed to differ statistically on other scores. CONCLUSION: These results, if replicated, could lead to identification of those at risk for anxiety and PTSD and to targeted interventions. PMID- 16259542 TI - Reduction of opioid-withdrawal symptoms with quetiapine. AB - OBJECTIVE: To determine the utility of quetiapine in a population undergoing ambulatory detoxification from opioids. METHOD: Medications utilized in our outpatient clinic for opioid withdrawal were evaluated for quality-assurance purposes. The treatment regimen generally included clonidine, hydroxyzine, trazodone, diphenoxylate/atropine, and sometimes chlordiazepoxide. Patients were also initially given eight 25-mg tablets of quetiapine and instructed to take 1 or 2 tablets every 4 hours as needed for symptoms of withdrawal or craving (with a maximum daily dose of 200 mg). Data were based on patient evaluations from June 2003 to June 2004. RESULTS: 41% of all patients (N = 213) successfully completed the detoxification phase of the program (i.e., completed at least 5 days of abstinence). A medication questionnaire was instituted for quality-assurance purposes after some apparent initial success with quetiapine. A retrospective analysis of these data revealed that, of the 107 patients evaluated for medication response, 79 reported that quetiapine helped reduce craving for opioids, 52 reported that quetiapine helped reduce their anxiety, 24 reported a reduction in somatic pain, 22 reported that quetiapine helped alleviate insomnia, and 14 reported an improved appetite. Four individuals did not feel quetiapine had any benefit, and another 7 were unable to tolerate quetiapine because of side effects. The quetiapine dose used ranged from 25 to 600 mg/day (mean +/- SD dose = 206 +/- 122 mg/day). CONCLUSIONS: Quetiapine use during opioid cessation was found to help abate symptoms of opioid withdrawal in our patient population and was generally well tolerated. PMID- 16259544 TI - Open-label study of atypical neuroleptic quetiapine for treatment of borderline personality disorder: impulsivity as main target. AB - BACKGROUND: Recent studies indicate that atypical neuroleptics may be safe and useful in treating many symptoms of borderline personality disorder (BPD), including impulsivity, which can constitute the core dimension of this pathology. This study aimed to evaluate the efficacy and safety of quetiapine in patients with well-defined BPD. It was hypothesized that quetiapine would reduce impulsivity (primary hypothesis) and also affective and micropsychotic symptoms, resulting in improved social and global functioning (secondary hypothesis). METHOD: Twenty-three outpatients with BPD according to DSM-IV criteria and the revised Diagnostic Interview for Borderlines completed a 12-week open-label study with quetiapine. The study was conducted from May 2001 to May 2003. The clinical efficacy was assessed using the following: Hamilton Rating Scales for Depression and Anxiety, Hopelessness Scale, Brief Psychiatric Rating Scale, Barratt Impulsivity Scale, Buss-Durkee Hostility Inventory, Temperament and Character Inventory, Social Adjustment Scale, and Global Assessment of Functioning. RESULTS: The mean daily dose of quetiapine (251 +/- 50 mg; range, 175-400 mg) was well tolerated. Impulsivity was significantly improved by quetiapine (p = .0015), as were most of our outcome measures: hostility, depression, anxiety, character dimensions, and social and global functioning (p < .05). In the small subgroup of patients with psychotic symptoms at baseline, there was a significant reduction in these symptoms (N = 8, p = .018). CONCLUSION: In a sample of patients with severe BPD without or with only few psychotic symptoms, a low dose of quetiapine was associated with a strong positive clinical impact, including improvement of impulsivity. PMID- 16259543 TI - Olanzapine/fluoxetine combination for treatment-resistant depression: a controlled study of SSRI and nortriptyline resistance. AB - BACKGROUND: This 8-week, double-blind, multicenter study was undertaken to replicate, in a larger sample of patients with treatment-resistant major depressive disorder (MDD; DSM-IV criteria), the results of a pilot study of the olanzapine/fluoxetine combination. METHOD: The study was begun in August 1999. The primary entry criterion was a history of failure to respond to a selective serotonin reuptake inhibitor (SSRI). Patients (N = 500) who subsequently failed to respond to nortriptyline during an open-label lead-in phase were randomly assigned to 1 of 4 treatment groups: olanzapine (6-12 mg/day) plus fluoxetine (25 50 mg/day) combination, olanzapine (6-12 mg/day), fluoxetine (25-50 mg/day), or nortriptyline (25-175 mg/day). The primary outcome measure was baseline-to endpoint mean change in score on the Montgomery-Asberg Depression Rating Scale (MADRS). RESULTS: At the 8-week study endpoint, MADRS total scores decreased by a mean 8.7 points from baseline (28.5) with the olanzapine/fluoxetine combination, 7.0 points from baseline (28.4) with olanzapine (p = .08), 8.5 points from baseline (28.4) with fluoxetine (p = .84), and 7.5 points from baseline (28.8) with nortriptyline (p = .30), with no significant differences among the therapies. The olanzapine/fluoxetine combination was associated with significantly (p < or = .05) greater improvement (decrease) in MADRS scores than olanzapine at weeks 2, 4, 6, and 7; than fluoxetine at weeks 2 through 5; and than nortriptyline at weeks 1 through 4. A post hoc analysis of a subgroup of patients who had an SSRI treatment failure during their current MDD episode (N = 314) revealed that the olanzapine/fluoxetine combination group had a significantly (p = .005) greater decrease in MADRS scores than the olanzapine group at endpoint. Safety data for the olanzapine/fluoxetine combination were similar to those for its component monotherapies. CONCLUSIONS: The olanzapine/fluoxetine combination did not differ significantly from the other therapies at endpoint, although it demonstrated a more rapid response that was sustained until the end of treatment. The results raised several methodological questions, and recommendations are made regarding the criteria for study entry and randomization. PMID- 16259545 TI - Predictors of everyday functioning among older Mexican Americans vs. Anglo Americans with schizophrenia. AB - OBJECTIVE: We assessed clinical, demographic, and cognitive predictors of everyday functioning in Mexican American and Anglo-American outpatients with schizophrenia. METHOD: Three groups of participants aged 40 years and over with a DSM-IV diagnosis of schizophrenia or schizoaffective disorder were compared: 79 Anglo-Americans, 24 Mexican American patients who chose to be tested in English, and 33 Mexican American patients who preferred Spanish. The study was conducted from October 2001 to July 2004. RESULTS: On demographic, clinical, cognitive, and functional measures, Anglo-American participants were generally similar to Mexican American participants who opted for being tested in English. Mexican American participants who preferred Spanish were significantly different from the other 2 groups on several measures, including everyday functioning performance. To determine the predictors of everyday functioning, separate regression analyses were conducted for each of the 3 groups. Cognitive ability consistently accounted for the greatest proportion of variance in performance regardless of ethnicity, language preference, or education. Among the Mexican American participants, level of acculturation was the second strongest predictor of everyday functioning when the group was examined as a whole; however, acculturation was not a significant predictor when controlling for language preference. CONCLUSION: Cognitive performance and language preference (a proxy for acculturation) may play a particularly important role in predicting ability to perform everyday tasks. Further studies to better understand the potential impact of ethnicity, culture, education, and language on everyday functioning may help develop more specific and culture-sensitive intervention strategies for different ethnic groups. PMID- 16259546 TI - Randomized trial of sertraline versus venlafaxine XR in major depression: efficacy and discontinuation symptoms. AB - BACKGROUND: The comparative efficacy of selective serotonin reuptake inhibitors (SSRIs) and serotonin-norepinephrine reuptake inhibitors (SNRIs) was recently debated. Meta-analyses, based mainly on fluoxetine comparator data, suggest that the SNRI venlafaxine has superior efficacy to SSRIs in treatment of major depression. OBJECTIVE: To compare quality of life (QOL), efficacy, safety, and tolerability associated with sertraline and venlafaxine extended release (XR) for treatment of DSM-IV major depression. METHOD: This was an 8-week, double-blind, randomized study of sertraline (50-150 mg/day) versus venlafaxine XR (75-225 mg/day), followed by a 2-week taper period. Subjects were recruited from 7 sites in Turkey and 6 sites in Australia between October 2002 and July 2003. The primary outcome measure was the Quality of Life Enjoyment and Satisfaction Questionnaire. Secondary outcome measures included measures of depression (including response and remission), anxiety, pain, safety (e.g., blood pressure), and tolerability (e.g., discontinuation symptoms). RESULTS: A total of 163 subjects received study treatment (women, 69%; mean age, 37.0 [SD = 12.9] years). No significant differences in QOL or efficacy were noted between treatments on the primary or secondary endpoints for the total study population or the anxious depression and severe depression subgroups. A priori analyses of symptoms associated with treatment discontinuation demonstrated no difference between treatment groups. However, in post hoc analyses, sertraline was associated with less burden of moderate to severe discontinuation symptoms. Venlafaxine XR was associated with a relative increase in mean blood pressure (supine diastolic blood pressure, -4.4 mm Hg difference at week 8/last observation carried forward). CONCLUSION: Sertraline and venlafaxine XR demonstrated comparable effects on QOL and efficacy in treatment of major depression, although sertraline may be associated with a lower symptom burden during treatment discontinuation and a reduced risk of blood pressure increase. PMID- 16259548 TI - Aripiprazole augmentation of selective serotonin reuptake inhibitors for treatment-resistant major depressive disorder. AB - BACKGROUND: Due to their favorable side effect profile, atypical antipsychotic agents offer important therapeutic advantages in mood disorders. Aripiprazole, an atypical antipsychotic agent with partial dopaminergic and serotonin 1A receptor agonist activity, may be particularly useful when used in conjunction with standard antidepressants in treatment-resistant depression. The purpose of this study was to test this hypothesis in depressed outpatients who have not experienced significant clinical improvement following an adequate trial of a selective serotonin reuptake inhibitor (SSRI). METHOD: 12 patients (mean +/- SD age = 46.6 +/- 11.3 years, 66.7% female) with major depressive disorder (MDD) diagnosed by use of the Structured Clinical Interview for DSM-IV-Axis I Disorders, who had failed to experience a clinical response following an adequate trial of an SSRI, were treated with open-label aripiprazole in addition to their SSRI for 8 weeks. Clinical response was defined as a 50% or greater decrease in depressive symptoms during the course of the trial (baseline-endpoint) as measured by the 17-item Hamilton Rating Scale for Depression total score. Data were collected from August 2003 to July 2004. RESULTS: 9/12 (75.0%) patients completed the trial. Using a completer analysis, 5/9 (55.6%) patients were classified as responders. An intent-to-treat (ITT) analysis resulted in 7 responders (58.3%). The overall proportion of remitters was 3/9 (33.3%) using a completer analysis and 5/12 (41.7%) using the ITT analysis. Aripiprazole administration appeared safe, with no severe adverse events observed in any of the study participants. CONCLUSIONS: These results suggest a possible augmentation role for aripiprazole when used in conjunction with SSRIs in SSRI resistant MDD. PMID- 16259547 TI - Adjunctive risperidone in generalized anxiety disorder: a double-blind, placebo controlled study. AB - OBJECTIVE: Although significant advances have been made in recent years in the treatment of generalized anxiety disorder (GAD), many patients remain symptomatic despite ongoing treatment, underscoring the need for adjunctive new treatments to help improve response. METHOD: Forty patients with a primary diagnosis of DSM-IV GAD, who continued to experience GAD symptoms despite current anxiolytic treatment of at least 4 weeks' duration, as evidenced by Hamilton Rating Scale for Anxiety (HAM-A) total score > or = 18 and Clinical Global Impressions Severity of Illness scale score of moderate or greater, completed a 1-week screening phase and were then randomly assigned to 5 weeks of double-blind adjunctive treatment with placebo or risperidone at flexible doses of 0.5 to 1.5 mg/day. Patients continued to take their anxiolytics throughout the study. The study was conducted from June 2001 through March 2003. RESULTS: Adjunctive risperidone was associated with statistically significant improvements in core anxiety symptoms, as demonstrated by greater reductions in HAM-A total scores (p = .034) and HAM-A psychic anxiety factor scores (p = .047) compared with placebo. Although change scores on other outcome variables, including response rates, were higher in the risperidone group, differences did not achieve statistical significance. CONCLUSION: Study findings suggest that risperidone at low doses may represent a useful tool in the management of symptomatic GAD patients. PMID- 16259549 TI - Metabolic side effects of second-generation antipsychotics in children and adolescents: a different story? PMID- 16259550 TI - Is there a real difference between the first onset of efficacy for atypical antipsychotic monotherapies in acute bipolar mania? PMID- 16259551 TI - Addition of lamotrigine to clozapine in inpatients with chronic psychosis. PMID- 16259552 TI - Thyroid dysfunction during treatment with atypical antipsychotics. PMID- 16259554 TI - Weight gain, sexual dysfunction, and bupropion. PMID- 16259556 TI - Treatment-emergent psychosis with aripiprazole. PMID- 16259557 TI - Silencing and variegation of gammaretrovirus and lentivirus vectors. AB - Retrovirus vectors integrate into the genome, providing stable gene transfer, but integration contributes in part to transcriptional silencing that compromises long-term expression. In the case of gammaretrovirus vectors based on murine leukemia virus, many integration events are completely silenced in undifferentiated stem cells and in transgenic mice. Gammaretrovirus vectors are also subject to variegation in which sister cells bearing the same provirus differentially express, and cell differentiation can lead to extinction of vector expression. In contrast, lentivirus vectors based on human immunodeficiency virus type 1 appear to express more efficiently, although other reports indicate that lentivirus vectors can be silenced. This review summarizes the key features of gammaretrovirus vector silencing. The evidence for and against gene silencing of lentivirus vectors is described with special emphasis on the potential effects of vector design, provirus copy number, and integration site preferences on silencing. This analysis suggests that the difference between selfinactivating (SIN) lentivirus vectors and their modified SIN gammaretrovirus counterparts may be less dramatic than previously thought. It will therefore be important to further characterize the mechanisms of silencing, in order to create better gammaretrovirus and lentivirus vectors that consistently express at single copy for gene therapy. PMID- 16259558 TI - Adeno-associated virus-mediated expression of vascular endothelial growth factor peptides inhibits retinal neovascularization in a mouse model of oxygen-induced retinopathy. AB - Vascular endothelial growth factor (VEGF) has been demonstrated to be a key stimulator of retinal neovascularization (NV), the most common cause of severe and progressive vision loss. In this study, we used a mouse model of oxygen induced retinopathy (OIR) to explore the potential of gene expression and secretion of short VEGF peptides as a treatment. Peptide-encoding fragments of exons 6 and 7 of the VEGF gene were cloned into a recombinant adeno-associated virus (rAAV) vector. Expression of each peptide in vector-injected eyes was confirmed by reverse transcription-polymerase chain reaction and Western blot analysis. Intravitreal injection of each rAAV vector inhibited retinal NV by 71 83% (p < 0.001) compared with contralateral control eyes in the OIR mouse. Injection and expression of these peptides did not seem to affect the normal appearance of the retina. The results demonstrated that exon 6- and 7-derived VEGF peptides effectively inhibited oxygen-induced retinal NV. Therefore, these VEGF peptides have potential in the treatment of angiogenesis-associated retinal diseases in humans. PMID- 16259559 TI - Induction of therapeutic antitumor immunity by in vivo administration of a lentiviral vaccine. AB - Direct in vivo administration of a lentiviral vaccine has been shown to transduce dendritic cells (DCs) in order to induce antigen-specific CD8+ T cell responses, but the efficacy of antitumor immunity has not been reported. In this study we tested whether direct in vivo administration of a lentiviral vaccine can induce selfantigen- based therapeutic antitumor immunity in murine tumor models. Lentiviral vector (LV) transduced DCs efficiently in vitro and was able to transduce DCs in vivo. LV-transduced DCs effectively presented antigens to T cells. Compared with a naked DNA tyrosinase-related protein-2 (TRP2)-heat shock protein-70 (hsp70) vaccine, the TRP2-specific interferon-gamma-producing CD8+ T cell response was augmented by direct in vivo administration of an LV-TRP2hsp70 vaccine, which induced significant therapeutic antitumor immunity in subcutaneous B16 and subcutaneous GL-26 models. Moreover, in vivo administration of an LV NeuEDhsp70 vaccine induced significant therapeutic antitumor immunity against spontaneous breast tumors in a BALB/c- Neu transgenic model. Our observations indicate that direct in vivo administration of a lentiviral vaccine not only enhances antigen-specific CD8+ T cell responses, but also generates significant therapeutic antitumor activities. PMID- 16259560 TI - Novel SRESPHP peptide mediates specific binding to primary medullary thyroid carcinoma after systemic injection. AB - The efficient and specific introduction of genes into cancer cells in vivo remains a major challenge for current gene therapy modalities. Peptides possess appropriate properties to serve as tumor-targeting agents. Thus, finding new cancer-selective peptides directing gene transfer to neoplastic cells by reducing transduction of normal cells is a central goal for molecular targeting. We have previously reported identification of a peptide (HTFEPGV) that selectively binds to human medullary thyroid carcinoma (MTC)-derived TT cells in vitro and transplanted tumor xenografts in vivo, using phage display. In the present study, we have performed this approach in primary orthotopically growing murine MTCs of RET-C634R transgenic mice as a clinically relevant model for thyroid cancer by intravenous injection of a complex peptide library. Two rounds of screening on primary tumors yielded multiple copies of a phage that displays a cyclic 7-amino acid peptide, SRESPHP, with a 3000-fold increase in titer between rounds 1 and 2. The selected phage showed highly specific binding to the tumor after systemic administration, whereas binding to other organs such as lung, liver, kidney, and heart was reduced up to 90%. After tail vein injection, homing to the tumor was substantially reduced in the presence of synthetic SRESPHP peptide, indicating that tumor phage interaction strictly depends on the displayed peptide. Immunohistochemical analysis of paraffin sections from mouse tissues revealed direct binding of the SRESPHP peptide to MTC tissue. Moreover, this peptide also mediates binding to human MTC cells in vitro and in vivo, suggesting abundant expression of its cognate receptor in murine and human medullary thyroid carcinoma. Because the SRESPHP peptide is also efficiently internalized into MTC cells, it likely provides the basis for a new selective therapy of medullary thyroid carcinoma. PMID- 16259561 TI - Gene therapy for lipoprotein lipase deficiency: working toward clinical application. AB - Lipoprotein lipase (LPL) deficiency causes hypertriglyceridemia and recurrent, potentially life-threatening pancreatitis. There currently is no adequate treatment for this disease. Previously, we showed that intramuscular administration of an adeno-associated virus serotype 1 (AAV1) vector encoding the human LPL(S447X) variant cDNA (AAV1-LPL(S447X)) normalized the dyslipidemia of LPL-/- mice for more than 1 year. In preparation for a clinical trial, we evaluated the safety and biodistribution of AAV1-LPL(S447X) in wild-type mice and fully characterized six LPL-deficient patients. Toxicological analysis in mice showed that intramuscular administration was well tolerated. Acute inflammatory response markers were transiently increased, and anti- AAV1 antibodies were generated. Histological analyses indicated a dose-dependent reversible spleen hyperplasia, and myositis at the injection sites. Biodistribution data showed short-term vector leakage from injection sites into the circulation, followed by liver-mediated clearance. Persistence of vector DNA was limited to the injected muscle and draining lymph nodes, and spread to reproductive organs was limited. Characterization of LPL-deficient patients showed that all patients presented with hypertriglyceridemia and recurrent pancreatitis. LPL catalytic activity was absent, but LPL protein levels were 20-100% of normal. Myoblasts derived from skeletal muscle biopsies of these patients were efficiently transduced by AAV1 LPL(S447X) and secreted active LPL. These data support the initiation of a clinical trial in LPL-deficient patients, for which regulatory approval has been granted. PMID- 16259562 TI - Novel compound enables high-level adenovirus transduction in the absence of an adenovirus-specific receptor. AB - Viral vectors are extensively used to deliver foreign DNA to cells for applications ranging from basic research to potential clinical therapies. A limiting step in this process is virus uptake and internalization into the target cells, which is mediated by membrane receptors. Although it is possible to modify viral capsid proteins to target the viruses, such procedures are complex and often unsuccessful. Here we present a rapid, inexpensive system for improving transduction of cells, including those that lack receptors for adenovirus fiber proteins. Addition of GeneJammer (Stratagene, La Jolla, CA) during the adenovirus transduction led to a significant increase in both the total number of transduced cells and the level of transgene expression per cell. Studies using cell lines deficient in adenovirus receptors demonstrated that addition of GeneJammer provided a novel cellular entry mechanism for the virus. These findings were tested in a cell-based gene therapy system for the induction of bone, which is contingent on high-level expression of the transgene. Inclusion of GeneJammer in either Ad5BMP2 or Ad5F35BMP2 transduction of a variety of cells demonstrated a correlating increase in bone formation. The results suggest a novel and versatile method for achieving high-level transduction using adenovirus. PMID- 16259563 TI - Comparative efficacy of intratracheal adeno-associated virus administration to newborn rats. AB - Transient local overexpression of genes that promote lung defense or repair may help to protect or promote alveolar development in premature neonates. We showed that the use of adenoviral vectors in neonates was limited by the induction of lung growth disorders. In the present work we compare the efficiency of gene transfer to the neonatal lung by three adeno-associated viral vectors: rAAV1, rAAV2, and rAAV5. Transduction efficiency was first measured in vitro, by infecting A549 immortalized human lung epithelial cells, and primary epithelial and mesenchymal cells isolated from human fetal lung. AAV vectors yielded similar low levels of luciferase gene expression in the different cell types. In vivo transduction efficiency was evaluated in newborn rats, with AAV-LacZ vectors being intratracheally instilled at 3 days of age. Both rAAV5 and rAAV1, but not rAAV2, induced significant lung beta-galactosidase expression, which persisted on day 35. Highest beta- galactosidase levels were measured with rAAV5, but remained far lower than those obtained with adenoviral vectors. A transient increase in alveolar macrophages was observed on day 6, but not on day 8, after rAAV5-LacZ instillation. Morphometric evaluation of lung structures was performed on day 21, and showed no altered lung growth. We conclude that rAAV1 or rAAV5 was more efficient at mediating gene transfer in the neonatal lung than was rAAV2, without adversely affecting lung development. However, in vivo transgene expression was relatively low, and needs to be improved for future therapeutic use of these adeno-associated vectors. PMID- 16259564 TI - Poly(ethylene imine)-poly(ethylene glycol) copolymers facilitate efficient delivery of antisense oligonucleotides to nuclei of mature muscle cells of mdx mice. AB - Antisense oligonucleotides (AO) can facilitate dystrophin expression via targeted exon skipping in cultured cells of Duchenne muscular dystrophy (DMD) patients and in the mouse model of DMD (mdx mice). However, the lack of effective means to deliver AO to myonuclei remains the foremost limitation to their usefulness in DMD gene therapy. In this study we show that copolymers of cationic poly(ethylene imine) (PEI) and poly(ethylene glycol) (PEG) facilitated efficient cellular uptake and nuclear delivery of AO in mature skeletal muscle fibers isolated from mdx mice. Confocal analysis of dual fluorescently tagged PEG-PEI-AO polyplexes, 24 hr after transfection, showed that the copolymer and AO were colocalized within punctate membrane- associated structures. Importantly, AO was efficiently translocated into myonuclei, whereas the copolymer was mostly excluded. The morphology of all transfected myofibers was perfectly maintained with no indication of damage or cytotoxicity. Quantitative fluorescence analysis showed that transfection with PEG-PEI-AO resulted in a 6-fold higher uptake of AO into myonuclei compared with transfections of AO alone. Interestingly, transfections with rhodamine-labeled PEG-PEI copolymers yielded an approximately 2- fold higher uptake of AO into myonuclei compared with transfections of unlabeled copolymers. Attempts to further increase AO delivery by addition of insulin-transferrin selenium (ITS) to the medium showed no further improvement in AO delivery. Dose response analysis indicated saturation of endocytotic uptake of the polyplex. Overall, we conclude that PEG-PEI copolymers represent high-capacity, nontoxic carriers for efficient delivery of AO to nuclei of mature myofibers. PMID- 16259565 TI - Amphiphilic block copolymers promote gene delivery in vivo to pathological skeletal muscles. AB - We reported that amphiphilic block copolymers hold promise as nonviral vectors for the delivery of plasmid DNA, ranging from 4.7 to 6.2 kb, to healthy muscle for the production of local or secreted proteins. To evaluate the efficiency of these vectors to deliver large plasmid DNA molecules to pathological muscles, plasmid DNAs of various lengths were complexed with Lutrol or poloxamine 304 and injected intramuscularly into dystrophic muscles. Lutrol-DNA and poloxamine 304 DNA complexes promoted gene transfer into muscles of the naturally occurring mouse model for DMD (mdx) in a dose- and plasmid DNA size-dependent manner. For small plasmid DNAs encoding reporter genes, this improvement over naked DNA was smaller in mdx than in the wild-type control strain. By contrast, Lutrol enabled us to deliver the large plasmid (16.1 kb) encoding the rod-deleted dystrophin in mdx mouse muscle, whereas the same amount of naked DNA did not lead to dystrophin expression, under the same experimental conditions. Lutrol-treated mdx mice showed the production of dystrophin in large numbers of muscle fibers. More importantly, we also found that expressing dystrophin with Lutrol led to restoration of the dystrophin-associated protein complex. Thus, we conclude that block copolymers constitute a novel class of vectors for the delivery of large plasmid DNA not only to healthy muscles but also to pathological muscle tissues. PMID- 16259566 TI - Dynamic gene expression after systemic delivery of plasmid DNA as determined by in vivo bioluminescence imaging. AB - Nonviral gene therapy approaches face the challenge of achieving stable gene expression in target organs and tissues. We used fluorescence microscopy and in vivo imaging after rapid, high-volume delivery of plasmid DNA (pDNA) encoding DsRed and luciferase as products of the same mRNA to detect localized gene expression in liver. Plasmids encoding the luciferase gene transcriptionally regulated by cellular and viral promoters were injected under similar conditions to test for potency and stability of gene expression in the liver of adult mice. Animals were imaged at 3, 6, 12, 24, and 48 hr and followed up at 1- to 2-week intervals over 2 months to identify maximum and persistent luciferase expression after injection of equimolar amounts of each plasmid. Emitted light representing luciferase expression was detected as early as 3 hr after pDNA infusion, reaching maximum levels at 12 hr for promoters containing viral sequences and at 24 hr for elements from human genes. Viral elements displayed 10- to 20-fold higher peak levels of expression but also yielded the most dramatic decline in expression over 8 weeks. In contrast, only a moderate decrease was observed for the cellular ubiquitin C promoter during that same period of time. Both promoter-deleted and phosphoglycerate kinase (PGK)-containing plasmids failed to maintain luciferase expression at levels above the limit of detection. These results demonstrate fine temporal analyses of reporter gene activity using promoters of various strength, suggesting an effective and reproducible method for studying gene therapy vectors in vivo. PMID- 16259567 TI - Effective growth arrest of human colon cancer in mice, using rat sodium iodide symporter and radioiodine therapy. AB - We have demonstrated that the rat sodium iodide symporter (rNIS) and 131I can effectively induce growth arrest of human prostate tumor xenografts [Mitrofanova, E., Unfer, R., Vahanian, N., Daniels, W., Roberson, E., Seregina, T., Seth, P., and Link, C. (2004). Rat sodium iodide symporter (rNIS) for radioiodide therapy of cancer. Clin. Cancer Res. 10, 6969-6976]. In that study the average size of tumors established in athymic nude mice was 200 +/- 50 mm3 when treated. Testing under more rigorous and extreme in vitro conditions will better evaluate the ability of an anticancer approach to induce tumor regression or killing capacity in preclinical studies. In this work the ability of the rNIS and 131I system to inhibit the growth of relatively large (about 800 mm3 when treated with 131I) and rapidly growing colon tumors in an animal model was examined. in vitro experiments demonstrated that transduction of human colon cancer cells with Ad rNIS resulted in a 100- to 150-fold increase in 125I uptake compared with nontransduced cells. Western blot analysis revealed robust expression of rNIS protein in cells 72-120 hr posttransduction with Ad-rNIS. Immunocytochemical analysis demonstrated that intracellular localization of rNIS-specific staining was observed mainly in plasma membranes of cells. in vitro studies revealed an immediate inhibition of growth of rapidly expanding tumors after radioiodine injection in the rNIS and 131I treatment group of mice. Twenty-seven percent of experimental mice survived more than 30 days (p = 0.019), whereas control groups had only 7% survival over 30 days. This is the first report demonstrating that rat NIS and 131I can effectively induce growth arrest of relatively large tumors in an animal model. PMID- 16259568 TI - Immunomodulation and protection induced by DNA-hsp65 vaccination in an animal model of arthritis. AB - We described a prophylactic and therapeutic effect of a DNA vaccine encoding the Mycobacterium leprae 65-kDa heat shock protein (DNA-hsp65) in experimental murine tuberculosis. However, high homology of the vaccine to the corresponding mammalian hsp60, together with the CpG motifs in the plasmidial vector, could trigger or exacerbate an autoimmune disease. In the present study, we evaluate the potential of DNA-hsp65 vaccination to induce or modulate arthritis in mice genetically selected for acute inflammatory reaction (AIR), either maximal (AIRmax) or minimal (AIRmin). Mice immunized with DNA-hsp65 or injected with the corresponding DNA vector (DNAv) developed no arthritis, whereas pristane injection resulted in arthritis in 62% of AIRmax mice and 7.3% of AIRmin mice. Administered after pristane, DNA-hsp65 downregulated arthritis induction in AIRmax animals. Levels of interleukin (IL)-12 were significantly lower in mice receiving pristane plus DNA-hsp65 or DNAv than in mice receiving pristane alone. However, when mice previously injected with pristane were inoculated with DNA hsp65 or DNAv, the protective effect was significantly correlated with lower IL-6 and IL-12 levels and higher IL-10 levels. Our results strongly suggest that DNA hsp65 has no arthritogenic potential and is actually protective against experimentally induced arthritis in mice. PMID- 16259569 TI - Serotype specificity of adenovirus purification using anion-exchange chromatography. AB - Recombinant adenoviruses continue to be a leading vector choice for gene transfer applications, with growing interest in the use of less prevalent serotypes, and of chimeras. As a result, the development of scaleable purification processes for alternative serotypes is needed. Anion-exchange chromatography is routinely used for scaleable adenovirus type 5 purification; however, retention varies for other serotypes because of differences in the exposed capsid proteins. Understanding how the viral surface influences retention behavior can provide a rational basis for chromatography development and optimization. In this work, chimeric vectors were used to show that the hexon protein is responsible for retention differences in anion-exchange chromatography. Next, the relative retention of eight serotypes from three subgroups was studied. Although all serotypes bound to the anion exchange resin, the sodium chloride required to elute the virus varied over a 2- fold range, from 270 to 490 mM. Retention was accurately correlated to the electrostatic properties of the hexon protein, with an average error in sodium chloride concentration required to elute of only 14 mM. This correlation enables preparative chromatography gradients for alternative serotypes to be established with minimal effort. PMID- 16259571 TI - Current pharmacotherapy for acromegaly: a review. AB - Acromegaly is associated with considerable morbidity and excess mortality; however, after effective treatment, both morbidity and mortality risks improve. Growth hormone excess in acromegaly can be controlled in many patients by pharmacotherapy alone, and with a combination of transsphenoidal surgery and pharmacotherapy in almost all patients. Since the clinical introduction of pegvisomant, a growth hormone-receptor antagonist, the role of radiotherapy is restricted. This review focuses on the treatment options for acromegaly (e.g., surgery, radiotherapy and pharmacotherapy with the depot preparations of the somatostatin analogues octreotide long-acting release formulation, lanreotide slow-release formulation and lanreotide Autogel, the growth hormone antagonist pegvisomant and the dopamine agonist cabergoline). Pharmacological characteristics of these drugs and the clinical and adverse effects are discussed individually and in relation to the other treatment modalities. The evidence for biochemical goals aimed at during medical treatment and the costs of pharmacotherapy are discussed. A new treatment algorithm is proposed, in which the choice between primary medical treatment and primary surgery is individualised, dependent on adenoma size and extension, patient factors (age, preference for therapy, contraindication for surgery), surgical experience of the centre and octreotide sensitivity of the adenoma. The high cost of lifelong medical treatment, especially of pegvisomant, must be weighed against the cost of a single surgical procedure. PMID- 16259572 TI - Management of adrenal insufficiency in different clinical settings. AB - Adrenal insufficiency is a rare disease, but its prevalence is increasing. The most frequent cause of primary adrenal insufficiency in western countries is autoimmune adrenalitis, whereas secondary adrenal insufficiency is most often caused by pituitary tumours and their treatment (e.g., surgery). Chronic glucocorticoid replacement consists of hydrocortisone 15-25 mg/day in divided doses and dose monitoring is largely based on clinical judgement. Fludrocortisone 0.05-0.2 mg/day is given for substitution in mineralocorticoid deficiency aiming at normotension, normokalaemia and a plasma renin activity in the upper normal range. It has recently been shown that, despite adequate glucocorticoid and mineralocorticoid replacement well being in patients with adrenal insufficiency is still impaired. Several studies have demonstrated that dehydroepiandosterone 25-50 mg/day p.o. may improve mood, fatigue, well-being and, in women, also sexuality, suggesting that dehydroepiandosterone should become part of the standard treatment regime. However, large Phase III trials of dehydroepiandosterone for adrenal insufficiency are still lacking and it has not yet been approved for the treatment of this disease. Patients with adrenal insufficiency are at risk of adrenal crisis, usually precipitated by major stress, such as severe infection or surgery. Early dose adjustments are required to cover the increased glucocorticoid demand in stress. Careful and repeated education of patients and their partners is the best strategy to avoid this life threatening emergency. Some recent studies suggest that during sepsis some patients with intact adrenal function may develop transient relative adrenal insufficiency and benefit from administration of hydrocortisone plus fludrocortisone. However, the pathophysiology and diagnosis criteria of relative adrenal insufficiency and its treatment remain unsettled issues. PMID- 16259573 TI - Insulin-sensitisers in the treatment of polycystic ovary syndrome. AB - Polycystic ovary syndrome (PCOS) is a common endocrine condition with reproductive and metabolic implications. In the current setting there is an evolving, yet inadequate, understanding of the pathophysiology, long-term health implications and ideal therapies for women with PCOS. Insulin resistance, secondary to both genetic and lifestyle factors, is integrally involved in the pathogenesis, the metabolic and clinical features and the long-term sequelae of PCOS in a majority of patients. Therapeutic strategies targeting insulin resistance ameliorate clinical features and may reduce long-term sequelae of PCOS, including diabetes. The main benefit of improved insulin resistance is to improve fertility and potentially to improve clinical features of hyperandrogenism and lower androgen levels. Insulin sensitisers also have the potential to delay the development of diabetes and cardiovascular disease in PCOS. Lifestyle therapy is indicated as the first intervention; however, metformin as an insulin sensitising agent has a role in first-line medical therapy in women with PCOS. Further research is needed to define the role of insulin sensitisers in PCOS and to determine the long-term risks and benefits of these therapies. PMID- 16259574 TI - Efficacy of the combination of an alpha1-blocker with an anticholinergic agent in the treatment of lower urinary tract symptoms associated with bladder outlet obstruction. AB - The combination of an alpha1-blocker with an anticholinergic is a new and promising therapeutic approach for bladder outlet obstruction and detrusor overactivity. Both placebo-controlled and comparative studies have demonstrated that the addition of an anticholinergic in the conventional treatment of patients with bladder outlet obstruction is safe, as the likelihood of acute urinary retention is low. Although the pathophysiology of detrusor overactivity is unknown and most probably multifactorial, it is not expected that the voiding phase is influenced by regular doses of anticholinergics, although high doses may affect detrusor contraction. However, safety issues must be studied further. The combination of tamsulosin with propiverine or tolterodine, and of doxasosin with tolterodine has been shown to cause a significant improvement of lower urinary tract symptoms when compared with alpha1-blocker monotherapy. Indisputably, the existing literature provides clear evidence that the combination of an alpha1 blocker with an anticholinergic extends physicians ability to manage lower urinary tract symptoms caused by bladder outlet obstruction and overactive bladder syndrome. PMID- 16259577 TI - Genetic polymorphisms of ATP-binding cassette transporters ABCB1 and ABCG2: therapeutic implications. AB - Pharmacogenomics, the study of the influence of genetic factors on drug action, is increasingly important for predicting pharmacokinetics profiles and/or adverse reactions to drugs. Drug transporters, as well as drug metabolism play pivotal roles in determining the pharmacokinetic profiles of drugs and their overall pharmacological effects. There is an increasing number of reports addressing genetic polymorphisms of drug transporters. However, information regarding the functional impact of genetic polymorphisms in drug transporter genes is still limited. Detailed functional analysis in vitro may provide clear insight into the biochemical and therapeutic significance of genetic polymorphisms. This review addresses functional aspects of the genetic polymorphisms of human ATP-binding cassette transporters, ABCB1 and ABCG2, which are critically involved in the pharmacokinetics of drugs. PMID- 16259575 TI - Drug therapy for the treatment of Langerhans cell histiocytosis. AB - Langerhans cell histiocytosis results from the abnormal accumulation of a class of dendritic cells normally found in the skin, which proliferate in many organ systems along with lymphocytes, macrophages and eosinophils. Standard therapy for Langerhans cell histiocytosis includes vinblastine and prednisone with or without methotrexate and mercaptopurine, depending on the extent of disease. Effective therapies for patients unresponsive to the above include cytosine arabinoside and cladribine. Thalidomide has proven useful for patients with Langerhans cell histiocytosis of the skin and/or bone. Emerging therapies include the use of monoclonal antibodies against the CD1a or CD52 epitopes found on Langerhans cells. Specific therapies directed against the cytokines that are apparently critical to the abnormal proliferation have not yet been defined. PMID- 16259576 TI - An update on the treatment options for gout and calcium pyrophosphate deposition. AB - Gout and calcium pyrophosphate deposition disease are two common causes of inflammatory joint disease. Despite differences underlying their pathogenesis, their clinical presentation and treatment share some common features. Optimal treatment for both requires prompt resolution of acute synovitis, reduction of chronic joint damage and management of associated conditions. Available therapeutic interventions and future strategies are reviewed in this article. PMID- 16259578 TI - Pharmacological advancements in the treatment of chronic anal fissure. AB - Chronic anal fissure is a tear in the lining of the anal canal that, if not treated appropriately at an early stage, causes considerable anal pain during defaecation. Surgery is no longer considered the first-line treatment of this common condition, as recent advancements in medical treatment has produced promising results in the healing of fissures, thus avoiding the unwanted complications that frequently occur following operative treatment. This review looks at those pharmacological agents used commonly in the treatment of chronic anal fissures and explores alternative therapies that may be of benefit in the future. PMID- 16259579 TI - The use of orlistat in the treatment of obesity, dyslipidaemia and Type 2 diabetes. AB - Orlistat (tetrahydrolipstatin) is an inhibitor of gastrointestinal lipases, especially pancreatic lipase. It is used as an adjunct to diet and exercise in order to achieve weight loss in obese individuals (body mass index > 30 kg/m2) or in overweight individuals (body mass index > 27 kg/m2) with other risk factors for atherosclerotic vascular disease, such as hypertension, dyslipidaemia or diabetes. Short- and long-term studies of up to 4 years duration have shown the drug to have significant benefits in weight loss, as well as in the reduction in lipids, glucose and haemoglobin A1c, and in time to onset of Type 2 diabetes compared with diet alone or placebo groups. The incremental amount of weight loss that orlistat produces is modest, but sufficient to result in improvement in obesity comorbidities such as elevated blood pressure, dyslipidaemia and hyperglycaemia compared with diet and exercise alone. Orlistat should only be prescribed for individuals who are motivated to adhere to lifestyle modifications, especially dietary fat restriction. PMID- 16259580 TI - Oseltamivir in the management of influenza. AB - Seasonal influenza causes significant morbidity and mortality in adults and children. However, a worldwide influenza pandemic could cause considerably more deaths (20-40 million) and would majorly disrupt everyday life in most countries. Oseltamivir has proven to be safe and effective for the prevention or treatment of all known influenza subtypes, reducing the severity and duration of symptoms, the complications arising from influenza infection (pneumonia, hospitalisation, antibiotic use) and mortality. Oseltamivir has been shown to be effective against pandemic strains of influenza, including the currently circulating strain (H5N1). As a result, the World Health Organization has recommended the stockpiling of oseltamivir in the event of an influenza pandemic. PMID- 16259581 TI - Review of immediate-release omeprazole for the treatment of gastric acid-related disorders. AB - Immediate-release omeprazole (Zegerid, Santarus) is the first immediate-release oral proton pump inhibitor to reach the market. As a powder formulation for oral suspension, it is indicated for the treatment of gastroesophageal reflux disease, erosive oesophagitis, duodenal ulcer and gastric ulcer, and is the only proton pump inhibitor approved for the reduction of risk of upper gastrointestinal bleeding in critically ill patients. Administration of immediate-release omeprazole at bedtime results in a rapid and sustained elevation of gastric pH, and seems to provide better night time control of gastric acidity than that observed with conventional morning dosing of delayed-release proton pump inhibitors. The immediate-release formulation may provide a good treatment option for patients who require flexible dosing, quick onset of action and nocturnal gastric acid control. PMID- 16259582 TI - Review of desloratadine for the treatment of allergic rhinitis, chronic idiopathic urticaria and allergic inflammatory disorders. AB - Desloratadine is a once-daily, non-sedating, non-impairing, selective histamine H1-receptor antagonist. It relieves the symptoms of seasonal allergic rhinitis (including nasal obstruction and congestion, and morning symptoms), perennial allergic rhinitis and chronic idiopathic urticaria by blocking multiple critical steps in the systemic allergic cascade and downregulating key allergy-induced inflammatory mediators. It also relieves asthma symptoms and decreases rescue medication use in patients with seasonal allergic rhinitis and comorbid asthma. Numerous clinical studies have demonstrated that desloratadine is safe, well tolerated and free of serious cardiac effects. Pharmacokinetic studies have demonstrated a low propensity for drug-drug or drug-food interactions. This review outlines the mechanism of action, efficacy and safety of desloratadine for the treatment of allergic inflammatory disorders. PMID- 16259583 TI - Budesonide and formoterol combination for the treatment of chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) is a major cause of morbidity and mortality throughout the world. Current guidelines recommend the addition of inhaled steroids to bronchodilators, which are central to the symptomatic management of COPD in patients with severe disease. Budesonide/formoterol is a combination inhaled steroid and long-acting bronchodilator delivered by a dry powder inhaler, approved for use in COPD. Two large, randomised, double-blind, 12 month studies found that combination budesonide/formoterol is more effective than either component alone in addressing many important aspects of the disease, such as pulmonary function, symptoms, use of relief medication, health-related quality of life and exacerbation in patients suffering from severe COPD. This review discusses the pharmacological and clinical properties of the drug. PMID- 16259584 TI - Pregabalin or morphine and gabapentin for neuropathic pain. PMID- 16259585 TI - Standardized experimental procedures in tissue engineering: cure or curse. PMID- 16259586 TI - Polymer scaffolds fabricated with pore-size gradients as a model for studying the zonal organization within tissue-engineered cartilage constructs. AB - The zonal organization of cells and extracellular matrix (ECM) constituents within articular cartilage is important for its biomechanical function in diarthroidal joints. Tissue-engineering strategies adopting porous three dimensional (3D) scaffolds offer significant promise for the repair of articular cartilage defects, yet few approaches have accounted for the zonal structural organization as in native articular cartilage. In this study, the ability of anisotropic pore architectures to influence the zonal organization of chondrocytes and ECM components was investigated. Using a novel 3D fiber deposition (3DF) technique, we designed and produced 100% interconnecting scaffolds containing either homogeneously spaced pores (fiber spacing, 1 mm; pore size, about 680 microm in diameter) or pore-size gradients (fiber spacing, 0.5 2.0 mm; pore size range, about 200-1650 microm in diameter), but with similar overall porosity (about 80%) and volume fraction available for cell attachment and ECM formation. In vitro cell seeding showed that pore-size gradients promoted anisotropic cell distribution like that in the superficial, middle, and lower zones of immature bovine articular cartilage, irrespective of dynamic or static seeding methods. There was a direct correlation between zonal scaffold volume fraction and both DNA and glycosaminoglycan (GAG) content. Prolonged tissue culture in vitro showed similar inhomogeneous distributions of zonal GAG and collagen type II accumulation but not of GAG:DNA content, and levels were an order of magnitude less than in native cartilage. In this model system, we illustrated how scaffold design and novel processing techniques can be used to develop anisotropic pore architectures for instructing zonal cell and tissue distribution in tissue-engineered cartilage constructs. PMID- 16259587 TI - Expansion of human chondrocytes in an intermittent stirred flow bioreactor, using modified biodegradable microspheres. PMID- 16259588 TI - Development of an ectopic site for islet transplantation, using biodegradable scaffolds. AB - Clinical islet transplantation in liver has achieved normoglycemia. However, this site may not be ideal for islet survival. To create a more optimal site for islet transplantation, we have developed a construct with biodegradable scaffolds. Islets were seeded in scaffolds and transplanted into the epididymal fat pad of diabetic BALB/c mice. Controls included islets transplanted underneath the kidney capsule or into the fat pad without scaffolds. All animals with islets in scaffolds or the kidney became normoglycemic and maintained this metabolic state. When islets were transplanted without scaffolds the time to achieve normoglycemia was significantly increased and less than 45% of mice survived. An oral glucose tolerance test was performed on the scaffold and kidney groups with similar blood glucose levels and area under the curve values between the groups. Grafts were removed at more than 100 days posttransplantation and all animals became hyperglycemic. There was no significant difference in insulin content between the grafts and all grafts were well vascularized with insulin-positive beta cells. Therefore, islets in scaffolds function and restore diabetic animals to normoglycemic levels, similar to islets transplanted underneath the kidney capsule, suggesting scaffolds can be used to create a site for islet transplantation. PMID- 16259589 TI - Improved microvascular network in vitro by human blood outgrowth endothelial cells relative to vessel-derived endothelial cells. AB - Evidence suggests that bone marrow-derived cells circulating in adult blood, sometimes called endothelial progenitor cells, contribute to neovascularization in vivo and give rise to cells expressing endothelial markers in culture. To explore the utility of blood-derived cells expressing an endothelial phenotype for creating tissue-engineered microvascular networks, we employed a three dimensional in vitro angiogenesis model to compare microvascular network formation by human blood outgrowth endothelial cells (HBOECs) with three human vessel-derived endothelial cell (EC) types: human umbilical vein ECs (HUVECs), and adult and neonatal human microvascular ECs. Under every condition investigated, HBOECs within collagen gels elongated significantly more than any other cell type. Under all conditions investigated, gel contraction and cell elongation were correlated, with HBOECs demonstrating the largest generation of force. HBOECs did not exhibit a survival advantage, nor did they enhance elongation of HUVECs when the two cell types were cocultured. Network formation of both HBOECs and HUVECs was inhibited by blocking antibodies to alpha2beta1, but not alpha(v)beta3, integrins. Taken together, these data suggest that superior network exhibited by HBOECs relative to vessel-derived endothelial cells is not due to a survival advantage, use of different integrins, or secretion of an autocrine/paracrine factor, but may be related to increased force generation. PMID- 16259590 TI - Proliferation and differentiation of rat bone marrow stromal cells on poly(glycolic acid)-collagen sponge. AB - We studied the effects of dexamethasone (Dex) and basic fibroblast growth factor (bFGF) on proliferation and differentiation of rat bone marrow stromal cells (RBMSCs), using three scaffolds: collagen sponge, poly(glycolic acid) (PGA) collagen sponge, and PGA-collagen (UV) sponge. RBMSCs were seeded into the sponges, and cultured in primary medium, primary medium with Dex, and primary medium with bFGF and Dex. Three weeks after cultivation, we examined alkaline phosphatase (ALP) activity and cell number in the sponges, and also performed macroscopic, light microscopic, and scanning electron microscopic (SEM) observations. Collagen sponge shrank considerably, but PGA-collagen and PGA collagen (UV) sponges maintained most of their original shape. PGA-collagen (UV) sponge supplemented with bFGF and Dex together had the highest ALP activity and cell number, followed by PGA-collagen sponge. Although collagen sponge showed cell proliferation only on the surface, the other two sponges showed cell proliferation in the interior. SEM showed the best cell attachment to PGA collagen (UV) sponge in the presence of bFGF and Dex, followed by PGA-collagen sponge. In conclusion, PGA-collagen (UV) and PGA-collagen sponges proved to be much more useful as scaffolding for bone regeneration when combined with bFGF and Dex. PMID- 16259591 TI - Methods: a comparative analysis of radiography, microcomputed tomography, and histology for bone tissue engineering. AB - This study focused on the assessment of radiography, microcomputed tomography, and histology for the evaluation of bone formation in a 15.0-mm defect in the rabbit radius after the implantation of a tissue-engineered construct. Radiography was found to be useful as a noninvasive method for obtaining images of calcified tissue throughout the time course of the experiment. With this method, however, image quality was low, making it difficult to obtain precise information about the location and quantity of the bone formed. Microcomputed tomography was used to create three-dimensional reconstructions of the bone (25 microm resolution). These reconstructions allowed for greater spatial resolution than the radiography, but did not allow for imaging of the implanted scaffold material or the surrounding, nonmineralized tissue. To visualize all materials within the defect area at the cellular level, histology was used. Histological analysis, however, is a destructive technique that did not allow for any further analysis of the samples. Each technique examined here has its own advantages and limitations, but each yields unique information regarding bone regeneration. It is only through the use of all three techniques that complete characterization of the bone growth and tissue/construct responses after implantation in vivo. PMID- 16259592 TI - Development of a biologic prosthetic composite for cartilage repair. AB - At present there is no satisfactory treatment for deep osteochondral defects. Here we report the development of a biologic prosthetic composite containing periosteum from 2-month-old rabbits and a porous tantalum scaffold. When cultured under chondrogenic conditions, the composites form a robust hyaline-like cartilage outgrowth that is attached to the porous scaffold by fibrous tissue ingrowth. The mechanical properties of these composites are similar to those of normal osteochondral plugs after only 6 weeks in culture. Thus, porous tantalum scaffolds are compatible with the chondrogenic capacity of periosteum. We hypothesize that these periosteum-porous tantalum composites will be useful for the repair of major osteochondral defects. However, in vivo experiments using biological resurfacing of large osteochondral defects with a porous tantalum scaffold and autologous periosteal graft in animal models are necessary to further explore this possibility. The implications of a successful method for cartilage regeneration would be great in terms of the number of patients affected and the quality of life for each of those patients. PMID- 16259593 TI - Stimulated endothelial cell adhesion and angiogenesis with laminin-5 modification of expanded polytetrafluoroethylene. AB - Biomedical implants often exhibit poor clinical performance due to the formation of a periimplant avascular fibrous capsule. Surface modification of synthetic materials has been evaluated to accelerate the formation of functional microcirculation in association with implants. The current study used a flow mediated protein deposition system to modify expanded polytetrafluoroethylene (ePTFE) with a laminin-5-rich conditioned growth medium and with medium from which laminin-5 had been selectively removed. An in vitro model of endothelial cell adherence determined that laminin-5 modification resulted in significantly increased adhesion of human microvessel endothelial cells to ePTFE. In vivo studies evaluating the periimplant vascular response to laminin-5-treated samples indicated that absorption of laminin-5-rich conditioned medium supported accelerated neovascularization of ePTFE implants. A flow system designed to treat porous implant materials facilitates laminin-5 modification of commercially available ePTFE, resulting in increased endothelial cell adhesion in vitro and increased vascularization in vivo. PMID- 16259594 TI - Enhancement of gap junctional intercellular communication of normal human dermal fibroblasts cultured on polystyrene dishes grafted with poly-N isopropylacrylamide. AB - Technology developed to allow recovery of cells without enzyme treatment, involving a dish grafted with a thermoreactive polymer gel of poly-N isopropylacrylamide (PIPAAm), was found to significantly enhance gap junctional intercellular communication (GJIC) in normal human dermal fibroblasts (NHDF cells). NHDF cells were cultured for 4 days on PIPAAm-grafted dishes irradiated with various doses of electron beams, and GJIC was assayed by the scrape-loading dye transfer method. The area of dye transfer was greater in the PIPAAm-grafted dishes than in the control culture dishes, indicating that the PIPAAm-grafted dishes enhanced the GJIC of NHDF cells. Connexin-43 (Cx43) expression was analyzed because Cx43 is considered to be a main component of the gap junctional channel. PIPAAm-grafted dishes irradiated with 100, 250, or 500 kGy of electron beams showed significantly enhanced expression of Cx43-NP, Cx43-P1, and especially Cx43-P2. Enhanced expression of Cx43-P2, a functional transmembrane protein, may be related to the promotion of GJIC. These results suggest that the PIPAAm-grafted dish not only enables the enzyme-free recovery of a cell monolayer for use in the construction of a three-dimensional artificial tissue, but also significantly contributes to the enhancement of GJIC, which may partly promote tissue strength on the surface of the PIPAAm-grafted dish. PMID- 16259595 TI - Factors that influence transgene expression and cell viability on DNA-PEI-seeded collagen films. AB - Gene delivery from tissue-engineering devices has the potential to improve healing, but better regulation of the level and duration of gene expression is needed. We hypothesized that transgene expression could be controlled by varying the fabrication and soaking parameters used in making collagen- based gene delivery scaffolds. Collagen films were made from acid-insoluble type I bovine dermal collagen and seeded with plasmid DNA encoding firefly luciferase, complexed with polyethylenimine. By varying the thickness of the films, the volume of the DNA soak solution, and the pH of the DNA soak solution, and by cross-linking the films, we identified variable combinations that produce significantly different levels of cell number and transgene expression in L-929 cells in vitro. Increasing film thickness or soak volume increased overall reporter gene expression. Decreasing film thickness or soak volume decreased cell number but did not significantly change reporter gene expression per cell. Cross linking by ultraviolet irradiation (before adding the DNA) significantly decreased transgene expression, probably because of decreased swelling of the collagen film. These results suggest that collagen-based biomaterials may be designed and fabricated to induce, in a controlled fashion, various levels of cellularity and transgene expression. PMID- 16259596 TI - Rapid isolation, expansion, and differentiation of osteoprogenitors from full term umbilical cord blood. AB - There is an urgent clinical requirement for appropriate bone substitutes that can be used for the repair and regeneration of diseased or damaged skeletal tissues. Cell-sourcing limitations in particular have affected progress, largely because of the shortage of accessible tissues capable of yielding sufficient numbers of viable osteoprogenitor cells. Previous work has suggested that umbilical cord blood (UCB) contains circulating progenitor cells (mesenchymal stem cells) capable of osteogenic differentiation, although a comparable number of reports refute this claim. From a screen of more than 20 different culture conditions, we have identified an optimal, simple, and reliable technique to generate, from full term human UCB, stromal cells with the ability to undergo rapid osteogenic differentiation. By comparing different sorting and culture strategies, we demonstrated that early exposure of mononuclear UCB cells to medium conditioned by osteoblastic cells in the presence of osteogenic supplements and human plasma, markedly increased the frequency of stromal cell growth, the rate of osteogenic differentiation, and their attachment to and spreading on calcium phosphate scaffolds. These findings suggest that full-term UCB may act as an appropriate source of osteoprogenitor cells, which will impact significantly on the development of autologous tissue- engineered bone constructs. PMID- 16259597 TI - Effects of in vitro preculture on in vivo development of human engineered cartilage in an ectopic model. AB - We investigated whether, and under which conditions (i.e., cell-seeding density, medium supplements), in vitro preculture enhances in vivo development of human engineered cartilage in an ectopic nude mouse model. Monolayer-expanded adult human articular chondrocytes (AHACs) were seeded into Hyalograft C disks at 1.3 x 10(7) cells/cm3 (low density) or 7.6 x 10(7) cells/cm3 (high density). Constructs were directly implanted subcutaneously in nude mice for up to 8 weeks or precultured for 2 weeks before implantation. Preculture medium contained either transforming growth factor-beta1 (TGF-beta1, 1 ng/mL), fibroblast growth factor 2, and platelet-derived growth factor (proliferating medium) or TGF-beta1 (10 ng/mL) and insulin (differentiating medium). Both in vitro and after in vivo implantation, constructs derived by cell seeding at high versus low density and precultured in differentiating versus proliferating medium generated more cartilaginous tissues containing higher amounts of glycosaminoglycan and collagen type II and lower amounts of collagen type I, and with higher equilibrium moduli. As compared with direct implantation of freshly seeded scaffolds, preculture of AHAC-Hyalograft C constructs in differentiating medium, but not in proliferating medium, supported enhanced in vivo development of engineered cartilage. The effect of preculture was more pronounced when constructs were seeded at low density as compared with high density. This study indicates that preculture of human engineered cartilage in differentiating medium has the potential to provide grafts with higher equilibrium moduli and enhanced in vivo developmental capacity than freshly seeded scaffolds. These findings need to be validated in an orthotopic model system. PMID- 16259598 TI - Downregulation of human type III collagen gene expression by antisense oligodeoxynucleotide. AB - The amount of type III collagen is increased during the early healing stage following ligament and tendon injury. Concomitantly, the mechanical properties of the healing tissues are abnormal and the fibril diameters are homogeneously small. It is therefore thought that downregulating type III collagen gene expression after injury may be helpful in improving the quality of healing tissue. In the current study, the efficacy of using antisense oligodeoxynucleotides (ODNs) to downregulate type III collagen gene expression in human patellar tendon fibroblasts (HPTFs) was tested, with Lipofectamine reagent used to deliver the ODN. It was shown that the majority of HPTFs can efficiently uptake antisense ODN from as early as 1 h to as long as 3 days after delivery; also, one selected ODN can consistently inhibit human type III collagen gene expression at both the mRNA and protein levels. Reverse transcriptase-polymerase chain reaction results showed that the inhibitory effects by this ODN were significant at 1 day, as the type III collagen mRNA level was 38.9 +/- 19.6 and 42.8 +/- 28.1% of missense and sense controls, respectively (p < 0.05). At 3 days, these differences could no longer be observed (p >0.05), but the amount of type III collagen protein was significantly less than for missense and sense controls (31.7 +/- 5.5 and 25.3 +/- 5.3%, respectively; p < 0.05). At 5 days after the delivery, these differences in protein were no longer observed (p > 0.05). Immunohistochemical staining of the type III collagen confirmed these results. The findings of this study demonstrate that antisense ODN can downregulate type III collagen gene expression of tendon fibroblasts. Therefore, this approach offers the potential to explore the effect of the reduction of type III collagen in healing ligaments and tendons as a means to improve their mechanical properties. PMID- 16259599 TI - Formation of lung alveolar-like structures in collagen-glycosaminoglycan scaffolds in vitro. AB - The objective of this study was to investigate the histology of tissue formed when fetal rat lung cells were grown in a collagen-glycosaminoglycan (GAG) tissue engineering scaffold. The goal was the formation of lung histotypic structures in the tissue-engineering scaffolds in vitro. Achieving this goal would facilitate future investigations of the effects of selected scaffold design parameters on processes that may underlie aspects of lung regeneration in vivo. Lung cells were obtained from Sprague-Dawley rats after 16 and 19 days of gestation. These dissociated cells were seeded into type I collagen-chondroitin 6-sulfate matrices, 8 mm in diameter by 2 mm in thickness, cross-linked and sterilized by dehydrothermal treatment. Approximately 28 million cells were seeded into each spongelike sample. Histological and immunohistochemical studies were performed at termination periods of 2 days and 1, 2, and 3 weeks. The enzymatically dissociated 19-day gestation fetal rat lung cells formed and maintained alveolar like structures, 50-60 microm in diameter, in the collagen- GAG scaffold. A novel finding was that all of the cell-seeded scaffolds underwent cell-mediated contraction that appeared to be associated with the finding by immunohistochemistry of expression of alpha-smooth muscle actin in some cells. These results demonstrate the capability of dissociated lung cells to form lung histotypic structures in collagen-GAG tissue-engineering scaffolds in vitro. This culture system may be of value in facilitating exploration of strategies for preparing such scaffolds for the regeneration of lung tissue in vivo. PMID- 16259600 TI - Modifying the NH2 and COOH termini of aquaporin-5: effects on localization in polarized epithelial cells. AB - To reengineer polarized epithelial cell functions directly in situ, or ex vivo in the fabrication of an artificial organ, it is necessary to understand mechanisms that account for polarized membrane sorting. We have used the aquaporins (AQPs), a family of homotetrameric water channel proteins, as model membrane proteins for this purpose. AQP monomers contain six transmembrane-spanning domains linked by five interconnecting loops, with the NH2 and COOH termini residing in the cytosol. AQP5 is localized in the apical membranes of several different epithelia in vivo, and in stably transfected MDCK-II cells grown as a polarized monolayer. We wished to identify a structural region(s) within rat AQP5 (rAQP5) important for apical localization, and to study the MDCK-II cell localization of rAQP5s modified in either their NH2 or COOH terminus. We show that the NH2- terminal region does not play a major role in apical localization as deletion of the NH2 terminus produced a modified rAQP5 construct (AQP5-NT(del)) that was stably expressed and localized primarily to the apical membranes of MDCK-II cells. Attachment of a FLAG epitope to the NH2 terminus of AQP5 (AQP5(flag) construct) also did not perturb apical localization. In addition, we found that the exchange of NH2-terminal regions between rAQP5 and human AQP1 (hAQP1; a nonpolarized AQP isoform) produced a modified rAQP5 construct (AQP5-1NT) and a modified hAQP1 construct (AQP1-5NT), each of which localized as the parental AQP (apically, and to both apical and basolateral membranes, respectively). In contrast, we found that deletion of the COOH terminus resulted in a modified rAQP5 construct (AQP5 CT(del)) that was unstably expressed and localized to intracellular site(s) in MDCK-II cells. Substitution of the COOH terminus of AQP1 with the COOH terminus of AQP5 also produced a construct (AQP1-5CT) transiently expressed in intracellular compartment(s). However, substitution of the COOH terminus of rAQP5 with the COOH terminus of hAQP1 produced a modified rAQP5 construct (AQP5-1CT) that was stably expressed and localized to basolateral membranes, suggesting the loss of an apical targeting/retention signal from rAQP5, the gain of a basolateral targeting/retention signal from hAQP1, or a combination of these two possibilities. PMID- 16259601 TI - Impregnation of plasmid DNA into three-dimensional scaffolds and medium perfusion enhance in vitro DNA expression of mesenchymal stem cells. AB - This article describes the development of an in vitro culture system to enhance the expression of a plasmid DNA for mesenchymal stem cells (MSCs) by a combination of plasmid DNA impregnation into three-dimensional cell scaffolds and culture methods. Gelatin was cationized by introducing spermine to the carboxyl groups for complexation with the plasmid DNA. As the MSC scaffold, poly(glycolic acid) (PGA) fiber fabrics, collagen sponges, and collagen sponges reinforced by incorporation of PGA fibers were used. A complex of cationized gelatin and plasmid DNA encoding bone morphogenetic protein 2 (BMP-2) was impregnated into the scaffolds. Plasmid DNA was released from PGA-reinforced collagen sponge for longer than from the other scaffolds. MCS were seeded into each type of scaffold and cultured by static, stirring, and perfusion methods. When MSCs were cultured in PGA-reinforced sponge, the level of BMP-2 expression was significantly enhanced by perfusion culture compared with the other culture methods, and the time of expression was prolonged. Irrespective of the culture method, the expression level was significantly higher from plasmid DNA impregnated in scaffold than by plasmid DNA in medium. The alkaline phosphatase activity and osteocalcin content of MSCs cultured in PGA-reinforced sponge by the perfusion method were significantly higher compared with those of other methods, and a significantly higher amount of plasmid DNA internalized into MSCs was observed. We conclude that a combination of plasmid DNA-impregnated PGA-reinforced sponge and the perfusion method was promising to promote in vitro gene expression for MSCs. PMID- 16259602 TI - Perfusion culture enhances osteogenic differentiation of rat mesenchymal stem cells in collagen sponge reinforced with poly(glycolic Acid) fiber. AB - The objective of this study was to obtain fundamental knowledge about in vitro culture systems to enhance the proliferation and differentiation of mesenchymal stem cells (MSCs) in collagen sponge reinforced by the incorporation of poly(glycolic acid) (PGA) fiber. A collagen solution with PGA fiber homogeneously localized at PGA:collagen weight ratios of 0.67, 1.25, 2.5, and 5 was freezedried, followed by cross-linking of combined dehydrothermal, glutaraldehyde, and ultraviolet treatment. Scanning electron microscopy revealed that collagen sponges exhibited homogeneous and interconnected pore structures with an average size of 180 microm, irrespective of PGA fiber incorporation. When rat MSCs were seeded into collagen sponge with or without PGA fiber incorporation, more attached cells were observed in collagen sponge incorporating PGA fiber than in collagen sponge without PGA fiber incorporation, irrespective of the PGA:collagen ratio. The proliferation and osteogenic differentiation of MSCs in PGA-reinforced sponge at a weight ratio of 5 were greatly influenced by the culture method and growth conditions. Alkaline phosphatase (ALP) activity and osteocalcin content of MSCs cultured in PGA-reinforced sponge by the perfusion method became maximum at a flow rate of 0.2 mL/min, although they increased with culture time period. It may be concluded that appropriate perfusion conditions enable MSCs to positively improve the extent of proliferation and differentiation. PMID- 16259603 TI - Osteoblast differentiation with titania and titania-silica-coated titanium fiber meshes. AB - Two surface-reactive sol-gel coatings, namely titania (TiO2) and a mixture of titania and silica (TiSi), were applied to titanium fiber meshes. Differentiation of rat bone marrow stromal cells toward an osteogenic phenotype with coated and uncoated (cpTi) substrates was compared. The amount of DNA in cpTi and TiSi matrices did not increase after day 3, but with TiO2 matrices the amount increased for 7 days. The prolonged period of proliferation with TiO2 scaffolds resulted in a delay in alkaline phosphatase induction. However, osteocalcin incorporation into extracellular matrix by day 14 was greater with TiO2 scaffolds than with cpTi scaffolds. Calcium deposition was also greater with TiO2-coated substrates than with uncoated substrates. With the TiSi scaffolds osteocalcin production and mineralization were lower than with the cpTi scaffolds. The current study confirms our previous findings that titanium fiber mesh supports attachment, growth, and differentiation of rat bone marrow stromal cells. Furthermore, the osteogenic capacities of cell-scaffold constructs under cell culture conditions were increased with a sol-gel-derived titania coating, but not with a titania-silica coating. PMID- 16259604 TI - Poly(ethylene glycol) hydrogel system supports preadipocyte viability, adhesion, and proliferation. AB - The ultimate goal of this research is to develop an injectable cell-scaffold system capable of permitting adipogenesis to abrogate soft tissue deficiencies resulting from trauma, tumor resection, and congenital abnormalities. The present work compares the efficacy of photopolymerizable poly(ethylene glycol) and specific derivatives as a scaffold for preadipocyte (adipocyte precursor cell) viability, adhesion, and proliferation. Four variations of a poly(ethylene glycol) scaffold are prepared and examined. The first scaffold consists of poly(ethylene glycol) diacrylate, which is not susceptible to hydrolysis or enzymatic degradation. Preadipocyte death is observed over 1 week in this hydrogel configuration. Adhesion sites, specifically the laminin-binding peptide sequence YIGSR, were incorporated into the second scaffold to promote cellular adhesion as a prerequisite for preadipocyte proliferation. Preadipocytes remain viable in this scaffold system, but do not proliferate in this nondegradable hydrogel. The third scaffold system studied consists of poly(ethylene glycol) modified with the peptide sequence LGPA to permit polymer degradation by cell secreted collagenase. No adhesion peptide is incorporated into this scaffold system. Cellular proliferation is initially observed, followed by cell death. The previous three scaffold configurations do not permit preadipocyte adhesion and proliferation. In contrast, the fourth system studied, poly(ethylene glycol) modified to incorporate both LGPA and YIGSR, permits preadipocyte adherence and proliferation subsequent to polymer degradation. Our results indicate that a scaffold system containing specific degradation sites and cell adhesion ligands permits cells to adhere and proliferate, thus providing a potential cell-scaffold system for adipogenesis. PMID- 16259605 TI - Evaluation of proliferation and functional differentiation of LLC-PK1 cells on porous polymer membranes for the development of a bioartificial renal tubule device. AB - To develop a bioartificial renal tubule system using renal tubular cells and porous polymer membrane hollow fibers, long-term maintenance of a confluent monolayer and the functionally differentiated condition of cells is essential. We examined the proliferation and functional differentiation of LLC-PK1 (Lewis-lung cancer porcine kidney 1) cells on two types of membranes: polysulfone and cellulose acetate. Cell proliferation was significantly higher on the polysulfone membrane than on the cellulose acetate membrane, and was enhanced by coating the membranes with various extracellular matrices. Confluent monolayer formation of cells was observed on matrix-coated polysulfone membrane but not on matrix-coated cellulose acetate membrane within 1 week. Cell proliferation continued for 3 weeks after confluent monolayer formation. Messenger RNA (mRNA) expression of glucose transporters, indicators of the functional differentiation of the LLC-PK1 cells, was observed in the polysulfone and cellulose acetate membrane groups, but was not observed in the nonporous polystyrene plate group under subconfluent conditions. Expression of glucose transporters mRNA was maintained for 3 weeks after confluent monolayer formation. Polysulfone membrane is more suitable than cellulose acetate membrane for a bioartificial renal tubule system with regard to LLC-PK1 cell proliferation. Extracellular matrix coating of the membrane further improves cell proliferation. PMID- 16259606 TI - Hyaline cartilage regeneration using mixed human chondrocytes and transforming growth factor-beta1- producing chondrocytes. AB - The purpose of this study was to investigate the efficacy of cartilage regeneration when using a mixture of transforming growth factor-beta1 (TGF-beta1) producing human chondrocytes (hChon-TGF-beta1) and primary human chondrocytes (hChon) ("mixed cells"), compared with either hChon-TGF-beta1 or hChon cells alone. Specifically, mixed cells or hChon cells were first injected intradermally into the backs of immune-deficient nude mice to test the feasibility of cartilage formation in vivo. Both the mixed cells and the hChon-TGF-beta1 cells alone induced cartilage formation in nude mice, whereas hChon cells alone did not. To further test the efficacy of the cells in generating cartilage, an artificially induced partial thickness defect of the femoral condyle of a rabbit knee joint was loaded with hChon-TGF-beta1 cells with or without mixing additional untransduced hChon cells, and hyaline cartilage regeneration was observed at 4 or 6 weeks. The efficiency of complete filling of the defect and the quality of tissue generated after implanting were evaluated on the basis of a histological grading system modified from O'Driscoll et al. (J. Bone Joint Surg. 70A, 595, 1988). Significantly, mixed cells (14.2 +/- 0.9) produced significantly better results than hChon-TGF-beta1 (9.0 +/- 1.7) or hChon (8.0 +/- 1.8) cells alone. Histological and immunohistochemical staining of the newly repaired tissues produced after treatment with either mixed cells or hChon-TGF-beta1 cells alone showed hyaline cartilage- like characteristics. These results suggest that the implantation of mixed cells may be a clinically efficient method of regenerating hyaline articular cartilage. PMID- 16259607 TI - Selective enhancement of cytochrome p-450 activity in rat hepatocytes by in vitro heat shock. AB - We investigated the effect of heat shock on cytochrome P-450 activity in rat hepatocytes and report a significant, selective, and time-dependent enhancement of cytochrome P-450 activity in heatshocked hepatocytes. Stable long-term cultures of rat hepatocytes were heat shocked (42.5 degrees C) for 1 to 3 h and allowed to recover at 37 degrees C. Cytochrome P-450-dependent ethoxyresorufin O dealkylase (EROD) and benzyloxyresorufin O-dealkylase (BROD) activities were measured up to 48 h after heat shock treatment. In general, the optimal heat shock exposure time was between 2 and 3 h. BROD activity (induced by sodium phenobarbital) increased approximately 6-fold in hepatocytes heat shocked for 3 h in comparison with hepatocytes maintained at 37 degrees C. EROD activity (induced by 3-methylcholanthrene) increased 2-fold on exposure to heat shock for 2 h. The expression of inducible heat shock proteins Hsp70 and Hsp32 was verified by Western immunoblot analyses. In the absence of the appropriate inducer, heat shock treatment did not enhance cytochrome P-450 activity. Furthermore, enhanced P-450 enzyme activity was delayed for heat-shocked hepatocytes. It is hypothesized that heat shock treatment attenuates the negative effects triggered by the addition of the toxic inducers and possibly stabilizes the levels of cytochrome P-450 proteins. These results suggest that heat shock treatment may be used to enhance the functionality of hepatocytes, specifically, in bioartificial liver assist devices. PMID- 16259608 TI - Review: application of stem cells for vascular tissue engineering. AB - As the prevalence of vascular disease has continued to expand, the need for a suitable arterial replacement has prompted researchers to look beyond synthetic and autologous grafts toward the field of tissue engineering. Advances in vascular tissue engineering have utilized both mesenchymal and hematopoietic stem cells as a cell source in an attempt to create a fully engineered small-diameter graft. Stem cells offer enormous potential as a cell source because of their proliferative and growth potential, and the application of stem cell technology has far-reaching implications for future applications. The innovative use of stem cells for vascular tissue engineering has opened new possibilities for a fully engineered blood vessel. The purpose of this review is to summarize the current perspective on the use of stem cells for vascular tissue engineering. It focuses principally on the classes of stem cells used, techniques for differentiation scaffolding technology, and the successes and failures of models. PMID- 16259609 TI - Novel methodology for fabrication of tissue-engineered tubular constructs using magnetite nanoparticles and magnetic force. AB - Novel technologies for creating three-dimensional constructs with complex shapes would be highly useful in tissue engineering. In the present study, tubular structures were constructed using magnetic force. Magnetite nanoparticles in cationic liposomes were taken up by target cells. The magnetically labeled cells were seeded onto ultralow-attachment plates, and a magnet was placed under the wells. After 24 h of culture, the magnetically labeled cells formed a cell sheet. Subsequently, when a cylindrical magnet was rolled onto the cell sheet, the cell sheet was attracted to the magnet and formed a tube around it. The magnet was then removed, leaving behind a tubular structure. Two types of tissue were used to create tubular structures: urinary tissue, consisting of a monotypic urothelial cell layer; and vascular tissue, consisting of heterotypic layers of endothelial cells, smooth muscle cells, and fibroblasts. The present results suggest that this novel methodology using magnetite nanoparticles and magnetic force, which we have termed "magnetic force-based tissue engineering" (Mag-TE), is a promising approach to constructing tissue-engineered tubular structures. PMID- 16259610 TI - Fibrin-polyurethane composites for articular cartilage tissue engineering: a preliminary analysis. AB - In this study we investigated the use of a fibrin hydrogel to improve the potential of a polyurethane (PU) scaffold-based system for articular cartilage tissue engineering. PU-only ("no-fibrin") and PU-fibrin ("fibrin") composites were cultured for up to 28 days and analyzed for DNA content, glycosaminoglycan (GAG) content, type II collagen content, GAG release, and gene expression of aggrecan, collagen I, and collagen II. The use of fibrin allowed for higher viable cell-seeding efficiency (10% higher DNA content on day 2 in fibrin versus no-fibrin composites) and more even cell distribution on seeding, a more than 3 fold increase in the percentage of newly synthesized GAG retained in the constructs, and 2- to 6-fold higher levels of type II collagen and aggrecan gene expression through day 14. Addition of aprotinin to the medium inhibited fibrin degradation, most noticeably in the center of the constructs, but had little effect on biochemical composition or gene expression. Short-term mechanical compression (0-10% sinusoidal strain at 0.1 Hz for 1 h, applied twice daily for 3 days) doubled the rate of GAG release from the constructs, but had little effect on gene expression, regardless of the presence of fibrin. Although further work is needed to optimize this system, the addition of fibrin hydrogel to encapsulate cells in the stiff, macroporous PU scaffold is a step forward in our approach to articular cartilage tissue engineering. PMID- 16259611 TI - Fabrication and endothelialization of collagen-blended biodegradable polymer nanofibers: potential vascular graft for blood vessel tissue engineering. AB - Electrospun collagen-blended poly(L-lactic acid)-co-poly(epsilon-caprolactone) [P(LLA-CL), 70:30] nanofiber may have great potential application in tissue engineering because it mimicks the extracellular matrix (ECM) both morphologically and chemically. Blended nanofibers with various weight ratios of polymer to collagen were fabricated by electrospinning. The appearance of the blended nanofibers was investigated by scanning electron microscopy and transmission electron microscopy. The nanofibers exhibited a smooth surface and a narrow diameter distribution, with 60% of the nanofibers having diameters between 100 and 200 nm. Attenuated total reflectance-Fourier transform infrared spectra and X-ray photoelectron spectroscopy verified the existence of collagen molecules on the surface of nanofibers. Human coronary artery endothelial cells (HCAECs) were seeded onto the blended nanofibers for viability, morphogenesis, attachment, and phenotypic studies. Five characteristic endothelial cell (EC) markers, including four types of cell adhesion molecule and one EC-preferential gene (von Willebrand factor), were studied by reverse transcription-polymerase chain reaction. Results showed that the collagen-blended polymer nanofibers could enhance the viability, spreading, and attachment of HCAECs and, moreover, preserve the EC phenotype. The blending electrospinning technique shows potential in refining the composition of polymer nanofibers by adding various ingredients (e.g., growth factors) according to cell types to fabricate tissue-engineering scaffold, particularly blood vessel-engineering scaffold. PMID- 16259612 TI - Functional bone engineering using ex vivo gene therapy and topology-optimized, biodegradable polymer composite scaffolds. AB - Bone tissue engineering could provide an alternative to conventional treatments for fracture nonunion, spinal fusion, joint replacement, and pathological loss of bone. However, this approach will require a biocompatible matrix to allow progenitor cell delivery and support tissue invasion. The construct must also support physiological loads as it degrades to allow the regenerated tissue to bear an increasing load. To meet these complex requirements, we have employed topology-optimized design and solid free-form fabrication to manufacture biodegradable poly(propylene fumarate)/beta-tricalcium phosphate composites. These scaffolds were seeded with primary human fibroblasts transduced with an adenovirus expressing bone morphogenetic protein-7 and implanted subcutaneously in mice. Specimens were evaluated by microcomputed tomography, compressive testing, and histological staining. New bone was localized on the scaffold surface and closely followed its designed contours. Furthermore, the total stiffness of the constructs was retained for up to 12 weeks after implantation, as scaffold degradation and tissue invasion took place. PMID- 16259613 TI - Tissue-engineered hybrid tooth and bone. AB - Tooth loss accompanied by alveolar bone resorption presents a significant clinical problem. We have investigated the utility of a tissue-engineering approach to provide corrective therapies for tooth-bone loss. Hybrid tooth-bone tissues were bioengineered as follows. Tooth implants were generated from pig third molar tooth bud cells seeded onto polyglycolide (PGA) and polyglycolide colactide (PLGA) scaffolds, and grown for 4 weeks in the omenta of adult rat hosts. Bone implants were generated from osteoblasts induced from bone marrow progenitor cells obtained from the same pig, seeded onto PLGA fused wafer scaffolds, and grown for 10 days in a rotational oxygen-permeable bioreactor system. The tooth and bone implants were harvested, sutured together, reimplanted, and grown in the omenta for an additional 8 weeks. Histological and immunohistochemical analyses of the excised hybrid tooth-bone constructs revealed the presence of tooth tissues, including primary and reparative dentin and enamel in the tooth portion of hybrid tooth-bone implants, and osteocalcin and bone sialoprotein-positive bone in the bone portion of hybrid tooth-bone constructs. Collagen type III-positive connective tissue resembling periodontal ligament and tooth root structures were present at the interface of bioengineered tooth and bone tissues. These results demonstrate the utility of a hybrid tooth-bone tissue engineering approach for the eventual clinical treatment of tooth loss accompanied by alveolar bone resorption. PMID- 16259614 TI - Neural tissue engineering: a self-organizing collagen guidance conduit. AB - We report a novel implantable device that will deliver a tethered aligned collagen guidance conduit containing Schwann cells into a peripheral nerve injury site. Cells (Schwann cells and fibroblasts) incorporated into tethered rectangular collagen gels contracted and resulted in uniaxial alignment. This tissue-engineered construct was tested in three-dimensional culture and demonstrated the ability to guide neurite extension from dissociated dorsal root ganglia. A silicone tube was adapted to provide tethering sites for an implantable construct such that uniaxial cell-generated tension resulted in the formation of a bridge of aligned collagen fibrils, with a resident Schwann cell population. The potential of this device for surgical nerve regeneration was assessed in a 5-mm defect in a rat sciatic nerve model. Neural regeneration through this device was significantly greater than in controls, demonstrating that this system has potential both as a simple robust clinical implant and as a three-dimensional engineered tissue model. PMID- 16259615 TI - Brain-derived neurotrophic factor enhances periodontal tissue regeneration. AB - To address whether brain-derived neurotrophic factor (BDNF) could be involved in periodontal tissue regeneration, we examined the effects of BDNF on proliferation and the expression of bone (cementum)- related proteins (osteopontin, bone morphogenetic protein [BMP]-2, type I collagen, alkaline phosphatase [ALPase], and osteocalcin) in cultures of human periodontal ligament (HPL) cells, which are thought to be prerequisite for periodontal tissue regeneration, and on proliferation and angiogenesis in human endothelial cells. Furthermore, we examined the effect of BDNF on the regeneration of periodontal tissues in experimentally induced periodontal defects in dogs. BDNF elevated the expression of ALPase and osteocalcin mRNAs and increased the synthesis of osteopontin, BMP 2, and type I collagen DNA in HPL cells. BDNF stimulated mRNA expression of vascular endothelial growth factor-B and tenascin-X, and proliferation and angiogenesis in human endothelial cells. In vivo studies showed that BDNF stimulated the formation of new alveolar bone cementum and connective new fibers, which were inserted into the newly formed cementum and bone. BDNF also stimulated blood capillary formation. These findings suggest that the regulation of functioning of periodontal ligament cells and endothelial cells by BDNF results in the promotion of periodontal tissue regeneration. PMID- 16259620 TI - Binding of the C-terminal domain of the HIV-1 capsid protein to lipid membranes: a biophysical characterization. AB - The capsid protein, CA, of HIV-1 forms a capsid that surrounds the viral genome. However, recent studies have shown that an important proportion of the CA molecule does not form part of this capsid, and its location and function are still unknown. In the present work we show, by using fluorescence, differential scanning calorimetry and Fourier-transform infrared spectroscopy, that the C terminal region of CA, CA-C, is able to bind lipid vesicles in vitro in a peripheral fashion. CA-C had a greater affinity for negatively charged lipids (phosphatidic acid and phosphatidylserine) than for zwitterionic lipids [PC/Cho/SM (equimolar mixture of phosphatidylcholine, cholesterol and sphingomyelin) and phosphatidylcholine]. The interaction of CA-C with lipid membranes was supported by theoretical studies, which predicted that different regions, occurring close in the three-dimensional CA-C structure, were responsible for the binding. These results show the flexibility of CA-C to undergo conformational rearrangements in the presence of different binding partners. We hypothesize that the CA molecules that do not form part of the mature capsid might be involved in lipid-binding interactions in the inner leaflet of the virion envelope. PMID- 16259622 TI - Lack of correlation between the levels of soluble cytotoxic T-lymphocyte associated antigen-4 (CTLA-4) and the CT-60 genotypes. AB - BACKGROUND: Cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) plays a critical role in downregulation of antigen-activated immune response and polymorphisms at the CTLA-4 gene have been shown to be associated with several autoimmune diseases including type-1 diabetes (T1D). The etiological mutation was mapped to the CT60 A/G single nucleotide polymorphism (SNP) that is believed to control the processing and production of soluble CTLA-4 (sCTLA-4). METHODS: We therefore determined sCTLA-4 protein levels in the sera from 82 T1D patients and 19 autoantibody positive (AbP) subjects and 117 autoantibody negative (AbN) controls using ELISA. The CT-60 SNP was genotyped for these samples by using PCR and restriction enzyme digestion of a 268 bp DNA segment containing the SNP. Genotyping of CT-60 SNP was confirmed by dye terminating sequencing reaction. RESULTS: Higher levels of sCTLA-4 were observed in T1D (2.24 ng/ml) and AbP (mean = 2.17 ng/ml) subjects compared to AbN controls (mean = 1.69 ng/ml) with the differences between these subjects becoming significant with age (p = 0.02). However, we found no correlation between sCTLA-4 levels and the CTLA-4 CT-60 SNP genotypes. CONCLUSION: Consistent with the higher serum sCTLA-4 levels observed in other autoimmune diseases, our results suggest that sCTLA-4 may be a risk factor for T1D. However, our results do not support the conclusion that the CT-60 SNP controls the expression of sCTLA-4. PMID- 16259621 TI - Role of N-terminal residues in the ubiquitin-independent degradation of human thymidylate synthase. AB - Thymidylate synthase (TS) catalyses the reductive methylation of dUMP to form dTMP, a reaction that is essential for maintenance of nucleotide pools during cell growth. Because the enzyme is indispensable for DNA replication in actively dividing cells, it is an important target for cytotoxic drugs used in cancer chemotherapy, including fluoropyrimidines (e.g. 5-fluorouracil and 5-fluoro-2' deoxyuridine) and anti-folates (e.g. raltitrexed, LY231514, ZD9331 and BW1843U89). These drugs generate metabolites that bind to the enzyme's active site and inhibit catalytic activity, leading to thymidylate deprivation and cellular apoptosis. Ligand binding to TS results in stabilization of the enzyme and an increase in its intracellular concentration. Previously, we showed that degradation of the TS polypeptide is carried out by the 26 S proteasome in a ubiquitin-independent manner. Such degradation is directed by the disordered N terminal region of the TS polypeptide, and is abrogated by ligand binding. In the present study, we have verified the ubiquitin-independent nature of TS proteolysis by showing that a 'lysine-less' polypeptide, in which all lysine residues were replaced by arginine, is still subject to proteasome-mediated degradation. In addition, we have mapped the structural determinants of intracellular TS degradation in more detail and show that residues at the N terminal end of the molecule, particularly the penultimate amino acid Pro2, play an important role in governing the half-life of the enzyme. This region is capable on its own of destabilizing an evolutionarily distinct TS molecule that normally lacks this domain, indicating that it functions as a degradation signal. Interestingly, degradation of an intrinsically unstable mutant form of TS, containing a Pro-->Leu substitution at residue 303, is directed by C-terminal, rather than N-terminal, sequences. The implications of these findings for the control of TS expression, and for the regulation of protein degradation in general, are discussed. PMID- 16259623 TI - Comparison of the systemic inflammatory response syndrome between monomicrobial and polymicrobial Pseudomonas aeruginosa nosocomial bloodstream infections. AB - BACKGROUND: Some studies of nosocomial bloodstream infection (nBSI) have demonstrated a higher mortality for polymicrobial bacteremia when compared to monomicrobial nBSI. The purpose of this study was to compare differences in systemic inflammatory response and mortality between monomicrobial and polymicrobial nBSI with Pseudomonas aeruginosa. METHODS: We performed a historical cohort study on 98 adults with P. aeruginosa (Pa) nBSI. SIRS scores were determined 2 days prior to the first positive blood culture through 14 days afterwards. Monomicrobial (n = 77) and polymicrobial BSIs (n = 21) were compared. RESULTS: 78.6% of BSIs were caused by monomicrobial P. aeruginosa infection (MPa) and 21.4% by polymicrobial P. aeruginosa infection (PPa). Median APACHE II score on the day of BSI was 22 for MPa and 23 for PPa BSIs. Septic shock occurred in 33.3% of PPa and in 39.0% of MPa (p = 0.64). Progression to septic shock was associated with death more frequently in PPa (OR 38.5, CI95 2.9-508.5) than MPa (OR 4.5, CI95 1.7-12.1). Maximal SIR (severe sepsis, septic shock or death) was seen on day 0 for PPa BSI vs. day 1 for MPa. No significant difference was noted in the incidence of organ failure, 7-day or overall mortality between the two groups. Univariate analysis revealed that APACHE II score > or = 20 at BSI onset, Charlson weighted comorbidity index > or = 3, burn injury and respiratory, cardiovascular, renal and hematologic failure were associated with death, while age, malignant disease, diabetes mellitus, hepatic failure, gastrointestinal complications, inappropriate antimicrobial therapy, infection with imipenem resistant P. aeruginosa and polymicrobial nBSI were not. Multivariate analysis revealed that hematologic failure (p < 0.001) and APACHE II score > or = 20 at BSI onset (p = 0.005) independently predicted death. CONCLUSION: In this historical cohort study of nBSI with P. aeruginosa, the incidence of septic shock and organ failure was high in both groups. Additionally, patients with PPa BSI were not more acutely ill, as judged by APACHE II score prior to blood culture positivity than those with MPa BSI. Using multivariable logistic regression analysis, the development of hematologic failure and APACHE II score > or = 20 at BSI onset were independent predictors of death; however, PPa BSI was not. PMID- 16259625 TI - Is it worth offering a routine laparoscopic cholecystectomy in developing countries? A Thailand case study. AB - OBJECTIVE: The study aims to investigate whether laparoscopic cholecystectomy (LC) is a cost-effective strategy for managing gallbladder-stone disease compared to the conventional open cholecystectomy (OC) in a Thai setting. DESIGN AND SETTING: Using a societal perspective a cost-utility analysis was employed to measure programme cost and effectiveness of each management strategy. The costs borne by the hospital and patients were collected from Chiang Rai regional hospital while the clinical outcomes were summarised from a published systematic review of international and national literature. Incremental cost per Quality Adjusted Life Year (QALY) derived from a decision tree model. RESULTS: The results reveal that at base-case scenario the incremental cost per QALY of moving from OC to LC is 134,000 Baht under government perspective and 89,000 Baht under a societal perspective. However, the probabilities that LC outweighed OC are not greater than 95% until the ceiling ratio reaches 190,000 and 270,000 Baht per QALY using societal and government perspective respectively. CONCLUSION: The economic evaluation results of management options for gallstone disease in Thailand differ from comparable previous studies conducted in developed countries which indicated that LC was a cost-saving strategy. Differences were due mainly to hospital costs of post operative inpatient care and value of lost working time. The LC option would be considered a cost-effective option for Thailand at a threshold of three times per capita gross domestic product recommended by the committee on the Millennium Development Goals. PMID- 16259624 TI - Protocol for: Sheffield Obesity Trial (SHOT): a randomised controlled trial of exercise therapy and mental health outcomes in obese adolescents [ISRCNT83888112]. AB - BACKGROUND: While obesity is known to have many physiological consequences, the psychopathology of this condition has not featured prominently in the literature. Cross-sectional studies have indicated that obese children have increased odds of experiencing poor quality of life and mental health. However, very limited trial evidence has examined the efficacy of exercise therapy for enhancing mental health outcomes in obese children, and the Sheffield Obesity Trial (SHOT) will provide evidence of the efficacy of supervised exercise therapy in obese young people aged 11-16 years versus usual care and an attention-control intervention. METHOD/DESIGN: SHOT is a randomised controlled trial where obese young people are randomised to receive; (1) exercise therapy, (2) attention-control intervention (involving body-conditioning exercises and games that do not involve aerobic activity), or (3) usual care. The exercise therapy and attention-control sessions will take place three times per week for eight weeks and a six-week home programme will follow this. Ninety adolescents aged between 11-16 years referred from a children's hospital for evaluation of obesity or via community advertisements will need to complete the study. Participants will be recruited according to the following criteria: (1) clinically obese and aged 11-16 years (Body Mass Index Centile > 98th UK standard) (2) no medical condition that would restrict ability to be active three times per week for eight weeks and (3) not diagnosed with insulin dependent diabetes or receiving oral steroids. Assessments of outcomes will take place at baseline, as well as four (intervention midpoint) and eight weeks (end of intervention) from baseline. Participants will be reassessed on outcome measures five and seven months from baseline. The primary endpoint is physical self-perceptions. Secondary outcomes include physical activity, self-perceptions, depression, affect, aerobic fitness and BMI. PMID- 16259626 TI - Resource-oriented music therapy for psychiatric patients with low therapy motivation: protocol for a randomised controlled trial [NCT00137189]. AB - BACKGROUND: Previous research has shown positive effects of music therapy for people with schizophrenia and other mental disorders. In clinical practice, music therapy is often offered to psychiatric patients with low therapy motivation, but little research exists about this population. The aim of this study is to examine whether resource-oriented music therapy helps psychiatric patients with low therapy motivation to improve negative symptoms and other health-related outcomes. An additional aim of the study is to examine the mechanisms of change through music therapy. METHODS: 144 adults with a non-organic mental disorder (ICD-10: F1 to F6) who have low therapy motivation and a willingness to work with music will be randomly assigned to an experimental or a control condition. All participants will receive standard care, and the experimental group will in addition be offered biweekly sessions of music therapy over a period of three months. Outcomes will be measured by a blind assessor before and 1, 3, and 9 months after randomisation. DISCUSSION: The findings to be expected from this study will fill an important gap in the knowledge of treatment effects for a patient group that does not easily benefit from treatment. The study's close link to clinical practice, as well as its size and comprehensiveness, will make its results well generalisable to clinical practice. PMID- 16259627 TI - The Oxford hip score: the patient's perspective. AB - BACKGROUND: In the last 25 years, assessment of orthopaedic intervention has become patient focused, with the development of self-completion patient-centred outcome measures. The Oxford hip score (OHS) is a joint specific outcome measure tool designed to assess disability in patients undergoing total hip replacement (THR). Although the psychometric properties of the OHS have been rigorously examined, there is little research on the patient's perspective of the OHS. Therefore, the aim of this study is to assess whether the OHS is an adequate disability measure from the patient's perspective using qualitative analysis of annotations written on the OHS by patients. METHODS: In total, 276 orthopaedic patients completed an OHS between April 2004 and May 2005. One hundred and fifty six pre-operative patients listed for a THR completed the OHS during a pre admission assessment clinic, and 120 post-operative patients completed the OHS postally in the home setting. Patient's unprompted annotations in response to the questions on the OHS were recorded and grouped into thematic categories. RESULTS: In total, 46 (17%) patients made 52 annotations when completing the OHS. These annotations identified five main areas of difficulty that patients experienced: lack of question clarity (particularly concerning the use of aids), difficulty in reporting measurements of pain, restrictive and irrelevant questions, the influence of co-morbidities on responses, and double-barrelled questions. CONCLUSION: Although the OHS is a useful short tool for the assessment of disability in patients undergoing THR, this study identified several problem areas that are applicable to patient-centred outcome tools in general. To overcome these current limitations, further work is underway to develop a more individualised patient-centred outcome measure of disability for use in patients with osteoarthritis. PMID- 16259629 TI - Evaluation of the profile of alopecia areata and the prevalence of thyroid function test abnormalities and serum autoantibodies in Iranian patients. AB - BACKGROUND: The study aimed at evaluating the prevalence of thyroid function abnormalities in patients with alopecia areata (AA) and its association with other autoimmune diseases and various autoimmune antibodies. METHOD: We retrospectively analyzed medical records of 123 patients with AA. The main site of involvement, pattern, and extent of alopecia as well as presence of the similar disease in first-degree family members and serologic status of patients were recorded. RESULTS: Participating in the study were 57 males and 66 females (6 to 59 years old). In the majority of patients (69.9%) the disease was manifested in the first two decades of life. Patients with family members having alopecia were recorded in 24.4%. Thyroid function abnormalities were found in 8.9% of patients. Positive autoimmune antibodies were associated with AA in 51.4% of patients with no significant association between the severity and duration of disease and presence of these antibodies. CONCLUSION: The incidence of positive auto-immune antibodies in Iranian patients is higher than previous reports. Concerning the female:male ratio, thyroid function tests and the prevalence of alopecia in first-degree relatives, our results are compatible with previous data obtained from different ethnic populations. Previous reports documented that a greater severity and longer duration of AA were seen in the early onset forms; however our result are relatively different which could be explained by differences in genetic factors. PMID- 16259628 TI - The microglial "activation" continuum: from innate to adaptive responses. AB - Microglia are innate immune cells of myeloid origin that take up residence in the central nervous system (CNS) during embryogenesis. While classically regarded as macrophage-like cells, it is becoming increasingly clear that reactive microglia play more diverse roles in the CNS. Microglial "activation" is often used to refer to a single phenotype; however, in this review we consider that a continuum of microglial activation exists, with phagocytic response (innate activation) at one end and antigen presenting cell function (adaptive activation) at the other. Where activated microglia fall in this spectrum seems to be highly dependent on the type of stimulation provided. We begin by addressing the classical roles of peripheral innate immune cells including macrophages and dendritic cells, which seem to define the edges of this continuum. We then discuss various types of microglial stimulation, including Toll-like receptor engagement by pathogen associated molecular patterns, microglial challenge with myelin epitopes or Alzheimer's beta-amyloid in the presence or absence of CD40L co-stimulation, and Alzheimer disease "immunotherapy". Based on the wide spectrum of stimulus specific microglial responses, we interpret these cells as immune cells that demonstrate remarkable plasticity following activation. This interpretation has relevance for neurodegenerative/neuroinflammatory diseases where reactive microglia play an etiological role; in particular viral/bacterial encephalitis, multiple sclerosis and Alzheimer disease. PMID- 16259630 TI - The reliability, validity, and preliminary responsiveness of the Eye Allergy Patient Impact Questionnaire (EAPIQ). AB - BACKGROUND: The Eye Allergy Patient Impact Questionnaire (EAPIQ) was developed based on a pilot study conducted in the US and focus groups with eye allergy sufferers in Europe. The purpose of this study was to present the results of the psychometric validation of the EAPIQ. METHODS: One hundred forty six patients from two allergy clinics completed the EAPIQ twice over a two-week period during the fall and winter allergy seasons, along with concurrent measures of health status, work productivity, and utility. Construct validity, reliability (internal consistency and test-retest), concurrent, known-group, and clinical validities, and responsiveness of the EAPIQ were assessed. Known-group validity was assessed by comparing EAPIQ scale scores between patients grouped according to their self rating of ocular allergy severity (no symptoms, very mild, mild, moderate, severe, very severe). Clinical validity was assessed by assessing differences in EAPIQ scores between groups of patients rated by their clinician as non symptomatic, mild, moderate, and severe. RESULTS AND DISCUSSION: Results from the validation study suggested the deletion of 14 of 43 items (including embedded questions) that required patients to complete the percentage of time they were troubled by something (daily activity limitations/emotional troubles). These items yielded a significant amount of missing or inconsistent data (50%). The resulting factor analysis suggested four domains: symptoms, daily life impact, psychosocial impact, and treatment satisfaction. When included as separate scales, the symptom-bother and symptom-frequency scales were highly correlated (> 0.9). As a consequence, and due to superior discriminative validity, the symptom bother and frequency items were summed. All items met the tests for item convergent validity (item-scale correlation = 0.4). The success rate for item discriminant validity testing was 97% (item-scale correlation greater with own scale than with any other). The criterion for internal consistency reliability (alpha coefficient > or = 0.70) was met for all EAPIQ scales (range 0.89-0.93), as was the criterion for test-retest reliability (intraclass correlation [ICC] > or = 0.70). Largely moderate correlations between the scales of the EAPIQ and the mini Rhinoconjunctivitis Quality of Life Questionnaire (miniRQLQ) and low correlations with the Health Utilities Index 2/3 (HUI2/3) were indicative of satisfactory concurrent validity. The EAPIQ symptoms, Daily Life Impact, and Psychosocial Impact scales were able to distinguish between patients differing in eye allergy symptom severity, as rated by patients and clinicians, providing evidence of satisfactory known-group and clinical validities, respectively. Preliminary analyses indicated the EAPIQ Symptoms, Daily Life Impact, and Psychosocial Impact scales to be responsive to changes in eye allergies. CONCLUSION: Following item reduction, construct validity, reliability, concurrent validity, known-group validity, and preliminary responsiveness were satisfactory for the EAPIQ in this population of ocular allergy patients. PMID- 16259631 TI - Porcine circovirus type 2 (PCV2) causes apoptosis in experimentally inoculated BALB/c mice. AB - BACKGROUND: We have previously described microscopic and electron microscopic alterations in lymphoid organs of PCV2 inoculated mice as apoptosis. In this study we wanted to investigate the molecular pathogenetic mechanism of PCV2 induced apoptosis. Eight-week old BALB/c mice were either sham inoculated (control mice) or inoculated intraperitoneally (ip) and intranasally (in) with a single (sPCV mice) or multiple (mPCV mice) doses of PCV2. Four control mice and 4 sPCV mice were sacrificed 7, 14, 28 and 42 days post inoculation (PI). All 4 mPCV mice were sacrificed 42 days PI. Following necropsy, immunohistochemistry for caspase 3 and in-situ TUNEL assay were performed on sections of spleen, lymph nodes, thymus and ileum from control, sPCV and mPCV mice. In addition, total RNA was extracted from spleens of control, sPCV and mPCV mice for simultaneous detection and semiquantitation of bcl-2 homologues and various caspase mRNAs using a multiprobe RNase protection assay system. RESULTS: PCV2 replicated and was associated with apoptosis in spleens, lymph nodes and Peyer's patches of infected BALB/c mice. Upregulation of caspase 1, 2, 3, 6, 7, 8, 11 and 12 and upregulation for the transcripts of apoptosis inhibitors bcl-2, bcl-w and bcl-X and apoptosis promoters' bax, bak and bad was detected in spleens of sPCV and mPCV mice, but not control mice. Apoptosis was further confirmed by light and electron microscopic morphology as well as by positive TUNEL assay and detection of activated caspase 3. PCV2 nucleic acid was detected by in-situ hybridization in the nuclei and cytoplasm of such apoptotic cells. CONCLUSION: The data presented here support the hypothesis that PCV2 induces apoptosis mediated through the activation of caspases 8 and 3 in the spleens of infected mice. PMID- 16259632 TI - Reversal of experimental colitis disease activity in mice following administration of an adenoviral IL-10 vector. AB - Genetic deficiency in the expression of interleukin-10 (IL-10) is associated with the onset and progression of experimental inflammatory bowel disease (IBD). The clinical significance of IL-10 expression is supported by studies showing that immune-augmentation of IL-10 prevents inflammation and mucosal damage in animal models of colitis and in human colitis. Interleukin-10 (IL-10), an endogenous anti-inflammatory and immunomodulating cytokine, has been shown to prevent some inflammation and injury in animal and clinical studies, but the efficacy of IL-10 treatment remains unsatisfactory. We found that intra-peritoneal administration of adenoviral IL-10 to mice significantly reversed colitis induced by administration of 3% DSS (dextran sulfate), a common model of colitis. Adenoviral IL-10 (Ad-IL10) transfected mice developed high levels of IL-10 (394 +/- 136 pg/ml) within the peritoneal cavity where the adenovirus was expressed. Importantly, when given on day 4 (after the induction of colitis w/DSS), Ad-IL10 significantly reduced disease activity and weight loss and completely prevented histopathologic injury to the colon at day 10. Mechanistically, compared to Ad null and DSS treated mice, Ad-IL10 and DSS-treated mice were able to suppress the expression of MAdCAM-1, an endothelial adhesion molecule associated with IBD. Our results suggest that Ad-IL10 (adenoviral IL-10) gene therapy of the intestine or peritoneum may be useful in the clinical treatment of IBD, since we demonstrated that this vector can reverse the course of an existing gut inflammation and markers of inflammation. PMID- 16259633 TI - Differential signaling mechanisms regulate expression of CC chemokine receptor-2 during monocyte maturation. AB - BACKGROUND: Peripheral blood monocytes and monocyte-derived macrophages are key regulatory components in many chronic inflammatory pathologies of the vasculature including the formation of atherosclerotic lesions. However, the molecular and biochemical events underlying monocyte maturation are not fully understood. METHODS: We have used freshly isolated human monocytes and the model human monocyte cell line, THP-1, to investigate changes in the expression of a panel of monocyte and macrophage markers during monocyte differentiation. We have examined these changes by RT-PCR and FACS analysis. Furthermore, we cloned the CCR2 promoter and analyzed specific changes in transcriptional activation of CCR2 during monocyte maturation. RESULTS: The CC chemokine receptor 2 (CCR2) is rapidly downregulated as monocytes move down the macrophage differentiation pathway while other related chemokine receptors are not. Using a variety of biochemical and transcriptional analyses in the human THP-1 monocyte model system, we show that both monocytes and THP-1 cells express high levels of CCR2, whereas THP-1 derived macrophages fail to express detectable CCR2 mRNA or protein. We further demonstrate that multiple signaling pathways activated by IFN gamma and M-CSF, or by protein kinase C and cytoplasmic calcium can mediate the downregulation of CCR2 but not CCR1. CONCLUSION: During monocyte-to-macrophage differentiation CCR2, but not CCR1, is downregulated and this regulation occurs at the level of transcription through upstream 5' regulatory elements. PMID- 16259634 TI - Communication of bed allocation decisions in a critical care unit and accountability for reasonableness. AB - BACKGROUND: Communication may affect perceptions of fair process for intensive care unit bed allocation decisions through its impact on the publicity condition of accountability for reasonableness. METHODS: We performed a qualitative case study to describe participant perceptions of the communication of bed allocation decisions in an 18-bed university affiliated, medical-surgical critical care unit at Sunnybrook and Women's College Health Sciences Centre. Interviewed participants were 3 critical care physicians, 4 clinical fellows in critical care, 4 resource nurses, 4 "end-users" (physicians who commonly referred patients to the unit), and 3 members of the administrative staff. Median bed occupancy during the study period (Jan-April 2003) was 18/18; daily admissions and discharges (median) were 3. We evaluated our description using the ethical framework "accountability for reasonableness" (A4R) to identify opportunities for improvement. RESULTS: The critical care physician, resource nurse, critical care fellow and end-users (trauma team leader, surgeons, neurosurgeons, anesthesiologists) functioned independently in unofficial "parallel tracks" of bed allocation decision-making; this conflicted with the official designation of the critical care physician as the sole authority. Communication between key decision-makers was indirect and could exclude those affected by the decisions; notably, family members. Participants perceived a lack of publicity for bed allocation rationales. CONCLUSION: The publicity condition should be improved for critical care bed allocation decisions. Decision-making in the "parallel tracks" we describe might be unavoidable within usual constraints of time, urgency and demand. Formal guidelines for direct communication between key participants in such circumstances would help to improve the fairness of these decisions. PMID- 16259635 TI - Clinical and pathologic factors associated with survival in young adult patients with fibrolamellar hepatocarcinoma. AB - BACKGROUND: Fibrolamellar Carcinoma (FLC), a subtype of hepatocellular carcinoma (HCC), is a rare primary hepatic malignancy. Several aspects of the clinic features and epidemiology of FLC remain unclear because most of the literature on FLC consists of case reports and small cases series with limited information on factors that affect survival. METHODS: We did a retrospective analysis of the clinical and histological characteristics of FLC. We also determined the rate of cellular proliferation in biopsies of these tumors. We assessed whether these variables were associated with survival. RESULTS: We found 15 patients with FLC out of 174 patients with HCC (8.6%). Between patients with these neoplasms, we found statistically significant survival, age at onset, level of alpha fetoprotein, and an earlier stage of the disease. The 1, 3 and 5 year survival in patients with FLC was of 66, 40 and 26% respectively. The factors associated with a higher survival in patients with FLC were age more than 23 years, feasibility of surgical resection, free surgical borders, absence of thrombosis or invasion to hepatic vessels and the absence of alterations in liver enzymes. The size of the tumor, gender, cellular proliferation and atypia did not affect the prognosis. CONCLUSION: We concluded that FLC patients diagnosed before 23 years of age have worse prognosis than those diagnosed after age 23. Other factors associated with worse prognosis in this study are: lack of surgical treatment, presence of positive surgical margins, vascular invasion, and altered hepatic enzymes. PMID- 16259636 TI - The neuroblast and angioblast chemotaxic factor SDF-1 (CXCL12) expression is briefly up regulated by reactive astrocytes in brain following neonatal hypoxic ischemic injury. AB - BACKGROUND: Stromal cell-derived factor 1 (SDF-1 or CXCL12) is chemotaxic for CXCR4 expressing bone marrow-derived cells. It functions in brain embryonic development and in response to ischemic injury in helping guide neuroblast migration and vasculogenesis. In experimental adult stroke models SDF-1 is expressed perivascularly in the injured region up to 30 days after the injury, suggesting it could be a therapeutic target for tissue repair strategies. We hypothesized that SDF-1 would be expressed in similar temporal and spatial patterns following hypoxic-ischemic (HI) injury in neonatal brain. RESULTS: Twenty-five 7-day-old C57BL/J mice underwent HI injury. SDF-1 expression was up regulated up to 7 days after the injury but not at the later time points. The chief sites of SDF-1 up regulation were astrocytes, their foot processes along blood vessels and endothelial cells. CONCLUSION: The localization of SDF-1 along blood vessels in the HI injury zone suggests that these perivascular areas are where chemotaxic signaling for cellular recruitment originates and that reactive astrocytes are major mediators of this process. The associated endothelium is likely to be the site for vascular attachment and diapedesis of CXCR4 receptor expressing cells to enter the injured tissue. Here we show that, relative to adults, neonates have a significantly smaller window of opportunity for SDF-1 based vascular chemotaxic recruitment of bone marrow-derived cells. Therefore, without modification, following neonatal HI injury there is only a narrow period of time for endogenous SDF-1 mediated chemotaxis and recruitment of reparative cells, including exogenously administered stem/progenitor cells. PMID- 16259637 TI - Association of paternal age at birth and the risk of breast cancer in offspring: a case control study. AB - BACKGROUND: Older paternal age may increase the germ cell mutation rate in the offspring. Maternal age may also mediate in utero exposure to pregnancy hormones in the offspring. To evaluate the association between paternal and maternal age at birth with the risk of breast cancer in female offspring, a case-control study was conducted in Korea. METHODS: Histologically confirmed breast cancer cases (n = 1,011) and controls (n = 1,011) with no present or previous history of cancer, matched on year of birth and menopausal status, were selected from several teaching hospitals and community in Seoul during 1995-2003. Information on paternal and maternal ages and other factors was collected by interviewed questionnaire. Odds ratio (OR) and 95% confidence interval (95% CI) were estimated by unconditional logistic regression model adjusting for family history of breast cancer in 1st or 2nd degree relatives, and lifetime estrogen exposure duration. RESULTS: The risk of breast cancer significantly increased as the paternal age increased (p for trend = 0.025). The association was stronger after controlling for maternal age; women whose fathers were aged >or=40 years at their birth had 1.6-fold increased risk of breast cancer compared with fathers aged <30 years. This association was profound in breast cancer cases in premenopausal women (OR = 1.9, 95% CI = 1.12-3.26, for paternal aged >or=40 vs. <30) (p for trend = 0.031). Although the risk of breast cancer increased as maternal age increased up to the intermediate, and then reduced; the risks in women whose mother were aged 25-29, 30-34, and >or=35 yrs at birth compared to women whose mothers were aged <25 years, were 1.2, 1.4, and 0.8, respectively, the trend was not significant (p for trend = 0.998). CONCLUSION: These findings suggest that older paternal age increases the risk of breast cancer in their female offspring. PMID- 16259639 TI - Assessment of pulse rate variability by the method of pulse frequency demodulation. AB - BACKGROUND: Due to its easy applicability, pulse wave has been proposed as a surrogate of electrocardiogram (ECG) for the analysis of heart rate variability (HRV). However, its smoother waveform precludes accurate measurement of pulse-to pulse interval by fiducial-point algorithms. Here we report a pulse frequency demodulation (PFDM) technique as a method for extracting instantaneous pulse rate function directly from pulse wave signal and its usefulness for assessing pulse rate variability (PRV). METHODS: Simulated pulse wave signals with known pulse interval functions and actual pulse wave signals obtained from 30 subjects with a trans-dermal pulse wave device were analyzed by PFDM. The results were compared with heart rate and HRV assessed from simultaneously recorded ECG. RESULTS: Analysis of simulated data revealed that the PFDM faithfully demodulates source interval function with preserving the frequency characteristics of the function, even when the intervals fluctuate rapidly over a wide range and when the signals include fluctuations in pulse height and baseline. Analysis of actual data revealed that individual means of low and high frequency components of PRV showed good agreement with those of HRV (intraclass correlation coefficient, 0.997 and 0.981, respectively). CONCLUSION: The PFDM of pulse wave signal provides a reliable assessment of PRV. Given the popularity of pulse wave equipments, PFDM may open new ways to the studies of long-term assessment of cardiovascular variability and dynamics. PMID- 16259638 TI - Immunohistochemical evidence for an endocrine/paracrine role for ghrelin in the reproductive tissues of sheep. AB - BACKGROUND: The gut hormone, ghrelin, is involved in the neuroendocrine and metabolic responses to hunger. In monogastric species, circulating ghrelin levels show clear meal-related and body weight-related changes. The pattern of secretion and its role in ruminant species is less clear. Ghrelin acts via growth hormone secretagogue receptors (GHSR-1a) to alter food intake, fat utilization, and cellular proliferation. There is also evidence that ghrelin is involved in reproductive function. In the present study we used immunohistochemistry to investigate the presence of ghrelin and GHSR-1a in sheep reproductive tissues. In addition, we examined whether ghrelin and GHSR-1a protein expression is developmentally regulated in the adult and fetal ovine testis, and whether there is an association with markers of cellular proliferation, i.e. stem cell factor (SCF) and proliferating cell nuclear antigen (PCNA). METHODS: Antibodies raised against ghrelin and its functional receptor, GHSR-type 1a, were used in standard immunohistochemical protocols on various reproductive tissues collected from adult and fetal sheep. GHSR-1a mRNA presence was also confirmed by in situ hybridisation. SCF and PCNA immunoexpression was investigated in fetal testicular samples. Adult and fetal testicular immunostaining for ghrelin, GHSR-1a, SCF and PCNA was analysed using computer-aided image analysis. Image analysis data were subjected to one-way ANOVA, with differences in immunostaining between time points determined by Fisher's least significant difference. RESULTS: In adult sheep tissue, ghrelin and GHSR-1a immunostaining was detected in the stomach (abomasum), anterior pituitary gland, testis, ovary, and hypothalamic and hindbrain regions of the brain. In the adult testis, there was a significant effect of season (photoperiod) on the level of immunostaining for ghrelin (p < 0.01) and GHSR-1a (p < 0.05). In the fetal sheep testis, there was a significant effect of gestational age on the level of immunostaining for ghrelin (p < 0.001), GHSR-1a (p < 0.05), SCF (p < 0.05) and PCNA (p < 0.01). CONCLUSION: Evidence is presented for the presence of ghrelin and its receptor in various reproductive tissues of the adult and fetal sheep. In addition, the data indicate that testicular expression of ghrelin and its receptor is physiologically regulated in the adult and developmentally regulated in the fetus. Therefore, the ghrelin ligand/receptor system may have a role (endocrine and/or paracrine) in the development (cellular proliferation) and function of the reproductive axis of the sheep. PMID- 16259640 TI - Stakeholder views regarding cultural diversity teaching outcomes: a qualitative study. AB - BACKGROUND: Cultural diversity teaching is increasingly present in both undergraduate and postgraduate training programmes. This study explored the views of stakeholders in medical education about the potential outcomes of cultural diversity teaching and how they thought cultural diversity programmes might be effectively evaluated. METHODS: A semi-structured interview was undertaken with 61 stakeholders (including policymakers, diversity teachers, students and users). The data were analysed and themes identified. RESULTS: Many participants felt that clinical practice was improved through 'cultural diversity teaching' and this was mostly as a result of improved doctor-patient communication. There was a strong view that service users need to participate in the evaluation of outcomes of cultural diversity teaching. CONCLUSION: There is a general perception, rather than clear evidence, that cultural diversity teaching can have a positive effect on clinical practice. Cultural diversity teaching needs to be reviewed in undergraduate and postgraduate medicine and better evaluation tools need to be established. PMID- 16259641 TI - Determination of Interleukin-6 and Tumor Necrosis Factor-alpha concentrations in Iranian-Khorasanian patients with preeclampsia. AB - BACKGROUND: Our objective was to determine the role of Interleukin-6 (IL-6) and Tumor Necrosis Factor-alpha (TNF-alpha), markers of immune activation and endothelial dysfunction, in patients with preeclampsia. METHODS: Twenty four women with preeclampsia and eighteen antepartum normotensive pregnant women were recruited as controls. Serum levels of IL-6 and TNF-alpha were measured by enzyme linked immunosorbent assay. We used independent-samples t test to assess the differences in the concentration of cytokines in preeclamptic patients and control subjects. RESULTS: IL-6 levels [mean (S.D.)] were significantly higher in preeclamptic women [5.8 (4.85) pg/ml] compared to normal pregnant women [3.01 (2.45) pg/ml] (p = 0.02). There was no significant change in concentration of TNF alpha in preeclamptic women [53.8 (30.0) pg/ml] compared to normal pregnant women [51.9 (33.8) pg/ml] (p > 0.1). CONCLUSION: The results of this study show that IL 6 as a pro-inflammatory cytokine is present in higher concentration in women with preeclampsia. The study was undertaken in women with established preeclampsia and it is not possible to determine whether the increased concentration of IL-6 is a cause or consequence of the disease. Furthermore, these findings suggest that serum TNF-alpha level is not associated with preeclampsia. PMID- 16259642 TI - Selecting additional tag SNPs for tolerating missing data in genotyping. AB - BACKGROUND: Recent studies have shown that the patterns of linkage disequilibrium observed in human populations have a block-like structure, and a small subset of SNPs (called tag SNPs) is sufficient to distinguish each pair of haplotype patterns in the block. In reality, some tag SNPs may be missing, and we may fail to distinguish two distinct haplotypes due to the ambiguity caused by missing data. RESULTS: We show there exists a subset of SNPs (referred to as robust tag SNPs) which can still distinguish all distinct haplotypes even when some SNPs are missing. The problem of finding minimum robust tag SNPs is shown to be NP-hard. To find robust tag SNPs efficiently, we propose two greedy algorithms and one linear programming relaxation algorithm. The experimental results indicate that (1) the solutions found by these algorithms are quite close to the optimal solution; (2) the genotyping cost saved by using tag SNPs can be as high as 80%; and (3) genotyping additional tag SNPs for tolerating missing data is still cost effective. CONCLUSION: Genotyping robust tag SNPs is more practical than just genotyping the minimum tag SNPs if we can not avoid the occurrence of missing data. Our theoretical analysis and experimental results show that the performance of our algorithms is not only efficient but the solution found is also close to the optimal solution. PMID- 16259643 TI - High carriage rate of high-level penicillin-resistant Streptococcus pneumoniae in a Taiwan kindergarten associated with a case of pneumococcal meningitis. AB - BACKGROUND: The Taiwan19F-14 Streptococcus pneumoniae clone and its variants are being found with increasing frequency in the Asia-Pacific region. A 5-year old child with S. pneumoniae meningitis caused by a high-level penicillin resistant strain (MIC = 4 microg/ml) was admitted to a hospital in southern Taiwan. We carried out a study to determine the potential source of this strain. METHODS: Nasopharyngeal cultures were obtained from all children attending the same kindergarten as the index case. To determine their relatedness all isolates were compared by serotype, antimicrobial susceptibility profile and pulsed field gel electrophoresis (PFGE). RESULTS: A high proportion of the children including the index case (32/78, 41.0%) carried S. pneumoniae in their nasopharynx (NP). The most common serotype was 19F (13/32, 40.6%). The PFGE types of the 19F serotype isolates obtained from the patient's blood, CSF and NP were identical and were related to 11 other serotype 19F NP isolates including 10 that were indistinguishable from the Taiwan19F-14 clone. All 14 isolates had similar high level penicillin and multi-drug resistance. The serotypes of the other 19 NP isolates included 6A (2), 6B (10), 23F (5), 9V (1) and 3 (1). The overall rate of penicillin resistance in these S. pneumoniae from these children was 87.5% (28/32), with an MIC50 of 2 and MIC90 of 4 ug/ml. In addition, multi-drug resistant-isolates (isolates resistant to 3 different classes of antimicrobials) accounted for 87.5% (28/32) of all isolates. CONCLUSION: The high carriage rate of high-level penicillin- and multi-drug- resistant S. pneumoniae in a kindergarten associated with a case of pneumococcal meningitis emphasizes the need for restraint in antibiotic use and consideration of childhood immunization with conjugate pneumococcal vaccine to prevent the further spread of resistant S. pneumoniae in Taiwan. PMID- 16259644 TI - TIMP3 mutation in Sorsby's fundus dystrophy: molecular insights. AB - Sorsby's fundus dystrophy (SFD) is a rare autosomal dominant disorder that results in degeneration of the macular region of the retina, with onset usually in the fourth to fifth decade of life. It leads to the rapid loss of central vision, often followed by further loss of peripheral vision. SFD shares several pathological features commonly found in the 'wet' or exudative form of age related macular degeneration (AMD), the most common cause of blindness in the elderly in developed countries. These phenotypic similarities have led to SFD being proposed as an acceptable genetic model for AMD. Whereas AMD appears to have a complex aetiology, with both genetic and environmental factors playing a role, SFD has been shown to be a single-gene disorder, linked to mutations in exon 5 of the tissue inhibitor of metalloproteinases 3 (TIMP3) gene on chromosome 22q12-q13. This review confines itself to a discussion of the known biochemical properties of the wild-type and SFD TIMP3 proteins and attempts to relate these to the pathology encountered in SFD patients. We also discuss briefly how, despite the lack of inherited mutations in the structural gene, the TIMP3 protein might play a role in the onset and progression of AMD. PMID- 16259645 TI - Efficacy and safety of 30 mg/d and 45 mg/d nemifitide compared to placebo in major depressive disorder. AB - Nemifitide is a novel pentapeptide antidepressant, which appears to be effective in the treatment of major depressive disorder (MDD). In the present study 81 patients with MDD, DSM-IV criteria were randomized following a 1-wk screening period to receive 30 mg/d nemifitide, 45 mg/d nemifitide or placebo in a 6-wk double-blind, multicentre, outpatient efficacy study. Nemifitide or placebo was delivered by subcutaneous injection for 2 wk daily for 5 days (Monday to Friday) in the first 2 wk and patients were followed up for a further 4 wk. The primary efficacy measure was the change from baseline on the Montgomery-Asberg Depression Rating Scale. Secondary measures included the 17-item Hamilton Psychiatric Rating Scale for Depression (HAMD), the CGI severity and improvement scale and the Carroll Self-Rating Scale for Depression. This proof-of-principle study demonstrated a statistically significant superiority of the 45-mg/d dose vs. placebo at the time-point of peak effect (1 wk after the end of treatment). There appeared to be a greater effect with the 45 mg/d nemifitide dose than with 30 mg/d. An additional exploratory analysis by stratification of all patients by severity above and below or equal to the median baseline HAMD score of 22 showed a higher percentage of responders for both doses of nemifitide with statistical separation from placebo for patients with baseline HAMD score of >22 (above the median). There was no significant difference among treatment groups for patients with baseline HAMD score of 0.05). Analysis of both irradiated and non-irradiated patients revealed that local and regional recurrence was determined independently by pathologic T (pT) stage, tumour localization, radiation status and pN stages (p < 0.05). The distant metastasis rate significantly depended on N stage and tumour localization (p < 0.05) and the rate of formation of multiple primary tumours was significantly affected by the patient's age and radiation status (p < 0.05). In conclusion irradiation of laryngeal cancer patients independently increases the risk of local and regional recurrence, and also increases the risk of multiple primary tumours while not significantly influencing the risk of distant metastasis. The risk of distant metastasis is affected by determinants of advanced lesions and tumour localization. PMID- 16259656 TI - Hyperbaric oxygen therapy for idiopathic sudden sensorineural hearing loss and tinnitus: a systematic review of randomized controlled trials. AB - BACKGROUND: Idiopathic sudden sensorineural hearing loss (ISSHL) and tinnitus are common. Hyperbaric oxygen therapy (HBOT) may improve hearing loss and/or reduce the intensity of tinnitus. METHODS: We performed a systematic search of the literature for randomized controlled trials, and made pooled analyses of pre determined clinical outcomes where possible. RESULTS: Six trials contributed to this review (304 subjects). Pooled analysis suggested a significantly increased chance of a 25 per cent improvement in hearing threshold on pure tone average with HBOT (relative risk (RR) 1.39, 95 per cent confidence interval (CI) 1.05 1.84, p = 0.02; number-needed-to-treat 5, 95 per cent CI 3-20), but not a 50 per cent increase (RR 1.53, 95 per cent CI 0.85-2.78, p = 0.16). The significance of any improvement in tinnitus following HBOT could not be assessed due to poor reporting. CONCLUSIONS: HBOT improved hearing, but the clinical significance of the level of improvement is not clear. Routine application of HBOT to patients with ISSHL is not justified by this review. More research is needed. PMID- 16259657 TI - Immunotherapy for allergic rhinitis--a United Kingdom survey and short review. AB - Allergic rhinitis (AR) is a common condition which is treated using different modalities, including immunotherapy. The aim of this study was to survey the current management strategies among ENT consultants in the UK in treating AR, and their views on immunotherapy. The study design was a postal questionnaire survey and the setting a university teaching hospital. Participants were consultant members of the British Association of Otolaryngologists - Head and Neck Surgeons (BAO-HNS). The main outcome measures were common treatment modalities adopted by the survey group to treat AR, and the number of consultants practising immunotherapy. The majority (81.1 per cent) of the consultants surveyed practise medical therapy with or without surgery. Immunotherapy is advised by 26 per cent of ENT consultants, but only 6.6 per cent currently administer immunotherapy. PMID- 16259659 TI - Sphenopalatine artery ligation: technical note. AB - Epistaxis is a common problem. Most patients presenting to hospital will stop bleeding with simple first-aid measures or with nasal packing. Those who do not stop will usually require surgical management. For persistent posterior epistaxis, the sphenopalatine artery may be ligated as the artery leaves the sphenopalatine foramen to enter the nasal mucosa of the lateral wall of the nose. This may be performed endoscopically. We describe the anatomy of the area and the surgical technique. We also present a brief review of the literature on this technique. PMID- 16259658 TI - The influence of the Internet on decision making in acoustic neuroma. AB - OBJECTIVE: To determine the access to and the use of the Internet by acoustic neuroma patients as a medical information source, and the influence of the Internet on decision-making, as well as the patients' views on the quality of particular sites. MATERIALS AND METHODS: A retrospective 27-item questionnaire was sent to 138 consecutive acoustic neuroma patients diagnosed between 1997 and 2002. The study included patients treated by conservative, radiotherapeutic and surgical modalities. MAIN OUTCOME MEASURES: Patient demographics, the extent of Internet use as well as quality and quantity of available information. RESULTS: There were 86.95 per cent questionnaires available for analysis. Twenty-four per cent accessed the Internet to obtain information about acoustic neuromas. Ninety seven per cent of patients said that the information provided in the clinic matched with the information obtained from the websites, 49 per cent said that the information obtained from the Internet did not influence decision making regarding acceptance of treatment of their tumour. Some patients said the Internet information was depressing, devastating, and they would never look up the Internet again for medical information. CONCLUSION: A significant proportion of our patients accessed the Internet for information. Caution should be exercised when advising patients regarding websites as the information on the Internet is often not controlled or checked for accuracy. Acoustic neuroma specialists and ENT surgeons in general should familiarize themselves with related websites so that patients can then be directed to high-quality sites. PMID- 16259660 TI - Self-retaining retractors in head and neck surgery. AB - The Norfolk and Norwich retractor is a vital tool in head and neck surgery. It is of great aid in training junior surgeons and has become an integral part of the standard neck dissection instrument set in our unit. This retractor enables good exposure of the carotid sheath, its atraumatic blunt tip retracting the carotid sheath without damage. It makes a single skin incision for neck exposure possible, rather than a Y, T or wine glass incision, avoiding a three-point junction, especially in the post-irradiated neck. In thyroid surgery it reduces the need for manual retraction thereby relieving the assistant surgeon and enhancing the quality of the learning experience. PMID- 16259661 TI - Pseudomonas cervical osteomyelitis with retropharyngeal abscess: an unusual complication of otitis media. AB - We present the unusual case of a 54-year-old diabetic man with chronic suppurative otitis media, presenting with cervical osteomyelitis and retropharyngeal abscess. This was treated with decompression, debridement and fusion from C2 to C4 with external halo-frame stabilization. Pseudomonas aeruginosa was cultured from the ear and the osteomyelitis specimen. Exploration of the left ear showed evidence of mucosal disease, with granulations in the middle ear and oedematous mucosa in the mastoid antrum, but no evidence of dural plate dehiscence. Haematogenous spread probably led to cervical osteomyelitis and retropharyngeal abscess formation. Cervical osteomyelitis may develop as a rare complication and present as a cause of severe neck pain in patients with otitis media. PMID- 16259663 TI - Consent and the use of Foley catheters in epistaxis. AB - The Medicines and Healthcare Products Regulatory Agency stated in 2003 that doctors should endeavour to avoid using products in treatments not covered by their product licence. Foley catheters are commonly used in the management of epistaxis although their product licence does not cover this. We undertook a questionnaire survey of members of the British Association of Otorhinolaryngologists--Head & Neck Surgeons to study the extent of the use of these catheters and the knowledge that members had of their legal status. Most members appear to use Foley catheters in the management of epistaxis; however, many are not aware that the product is not licensed for this purpose. Because of this lack of knowledge, only half obtain verbal consent for treatment with this device and only a very small number obtain written consent from patients. In the era of increasing litigation, documentation of informed consent could be considered mandatory to protect us from possible legal action, and this needs to be known by all practising otolaryngologists. PMID- 16259664 TI - Vocal fold injury following endotracheal intubation. AB - Vocal fold scarring results in the formation of fibrous tissue which disturbs the vibratory pattern of the fold during phonation. However, vocal fold scarring in humans is poorly understood because of the lack of clear case reports focusing on voice quality. The authors present a case of vocal fold scarring with changes in voice quality. At the time of injury the pedicle mucosa was cemented with fibrin glue. Phonation was inhibited for two weeks and tranilast (300 mg/day) was given for 3 months. Sixty-nine days later, perceptual evaluation showed a normal result and the phonation time became better, but the mucosal vibration was still lacking. Ninety-seven days later, mucosal vibration was finally restored. We suggest that characterization of vocal fold scarring in humans may be different from that in animals, and recommend that surgical management should be avoided for at least three months after injury. PMID- 16259662 TI - Airway obstruction: a rare complication of thrombolytic therapy. AB - Bleeding into head and neck structures as a complication of thrombolysis is extremely rare but potentially lethal. It is usually associated with local trauma. We describe a case of a 67-year-old woman who developed spontaneous, massive pan aerodigestive tract haemorrhage precipitating airway obstruction following administration of streptokinase for myocardial infarction; a previously unreported complication. This case highlights the importance of being vigilant of airway symptoms following such treatment, prompting early involvement of anaesthetic and otolaryngology personnel. PMID- 16259665 TI - A cavernous haemangioma of the thyroid gland. AB - Haemangioma of the thyroid gland is extremely rare. We report a case of a cavernous haemangioma of the neck that was located both inside and outside the thyroid gland of a 21-year-old man. Ultrasonography of the neck revealed numerous calcified nodules. Computed tomography (CT) showed a large calcified soft tissue mass in the right lobe of the thyroid. Based on the imaging findings a papillary thyroid carcinoma was suspected, but examination of the surgical specimen revealed a huge cavernous haemangioma containing numerous phleboliths. PMID- 16259666 TI - Laser arytenoidectomy for bilateral vocal fold palsy in Parkinson's disease. AB - We report a case of laser arytenoidectomy for bilateral abductor palsy of the vocal fold in a patient with Parkinson's disease. Parkinson's disease is known to be a rare cause for bilateral vocal fold palsy and this is the second case reported in the English literature. Majority of the reports to date are in the non-English literature. Tracheostomy has been the classic treatment option offered to these patients. To date no report has been found in the literature about a laser arytenoidectomy being performed in a patient suffering from bilateral vocal fold palsy in Parkinson's disease. We have suggested this option as it improves the quality of life of the individual and avoids the difficulty faced by a Parkinson's patient in managing tracheostomy care due to bradykinesia, rigidity and tremor. PMID- 16259667 TI - Colonic carcinoma metastasis to the thyroid: a case of skip metastasis. AB - Colonic carcinoma metastasis to the thyroid is rare. Here the authors present the case of an 81-year-old lady who presented with metastatic colonic adenocarcinoma in her thyroid gland. This case is unique as it is the first to demonstrate metastasis from the colon to the thyroid with no other site involvement. The use of cytokeratin immunohistochemical staining is reviewed along with the current perspectives on the concept of skip metastasis. PMID- 16259668 TI - Involuntary automaticity: a work-system induced risk to safe health care. AB - Automaticity is the term given by psychologists to the skilled action that people develop through repeatedly practising the same activity, for example driving a car. Usually, automaticity is discussed in terms of the benefits it brings to people, such as the reduction in the degree of conscious attention a person needs to pay to such skilled activities. However, there is evidence to suggest that substantial costs may also be associated with such learned behaviour. Managing patient safety is a difficult task and one of the ways in which health-care professionals seek to accomplish it is through the use of verbal challenge response protocols. However, it is argued in this paper that it is possible for the negative effects of automaticity to involuntarily capture those using such verbal checklist techniques and cause them to erroneously believe that the treatment that they are about to administer to a patient is safe when it is not. This phenomenon does not, however, seem to have been recognized by the health care community nationally or internationally. We conclude that patient safety could be significantly improved worldwide if the organizational arrangements which appear to induce involuntary automaticity were to be robustly addressed by the management of all health-care organizations. PMID- 16259669 TI - The nursing shortage and the scope for reducing it by redefining their role. AB - The objective of this research is to measure the extent to which nurses are undertaking tasks that could be left to support workers. The method employed was firstly to classify the activities being undertaken by nurses. This was achieved by surveying appropriate professional groups. Secondly, the amount of time nurses spent on each activity was measured. The results indicate that nurses spend at least 8% of their time undertaking activities which could be undertaken by less qualified staff. It was also found that 6% of unregistered nurses' time is spent on activities which are classified as being within the remit of a registered nurse. The scale of these results raises significant efficiency and effectiveness issues with respect to the use of both groups of staff and therefore has major implications for their management. PMID- 16259670 TI - Managing health inequalities locally: a baseline survey of primary care trusts' experience with health equity audit in the implementation year. AB - Health equity audit (HEA), a pragmatic policy tool to ensure that services and resources are focused on issues that have the highest impact on health inequalities, has now become embedded in the national strategy to tackle these inequalities through a new mandatory requirement that primary care trusts (PCTs) conduct one such audit annually. This study aimed to assess their experience through a national baseline survey in 2004, all PCTs (n = 303) being electronically mailed a questionnaire with non-respondent follow-up. Replies were received from 132 PCTs (44%), a representative sample of PCT diversity, most of whom had completed only a few steps in the audit cycle. Audit topics (most frequently coronary heart disease, smoking cessation, and access issues) and dimensions of inequity (mainly area and deprivation) were agreed through only limited engagement with local strategic partnerships. Local public health networks and multiagency teams were infrequent partners in undertaking the HEA. Most PCTs wanted comparator data, good practice examples, and specific methodological expertise. While significant progress has been made, this survey shows only limited use of HEA as a tool for multisectoral use by PCTs in partnership with others and a focus on intra-PCT comparisons at the expense of those with a wider pool of 'look-a-likes'. PMID- 16259671 TI - An analysis of high-cost users at an Australian public health service organization. AB - High-cost users generate extremely high costs when compared with average users in the same diagnostic-related group (DRG). They represent a major financial loss for a health service organization. The research was conducted using an area health service patient database for online analytical processing to produce descriptive statistics and graphs of 'high-cost' and 'non-high-cost users'. Trends and patterns were identified across key variables derived from clinical, financial and operational categories. The main results are: 20% of costs are spent by 3% of the population; elective admission is higher in the high-cost group; tracheostomy has the most number of cases and is the most expensive DRG; LOS is mostly longer for complex cases however, high costs can be attributed to other factors. In conclusion, these findings are potentially useful to patients, medical staff, management and health service decision-makers. The limitation of this study is the exclusion of profitability. PMID- 16259672 TI - End-of-life care volunteers: a systematic review of the literature. AB - This report presents a review of 1988 and onwards research and other literature on end-of-life (EOL) care volunteers. Only 18 research or case studies articles were identified for an integrative review through a search of nine library databases. A review of this literature revealed three themes: (1) the roles of EOL volunteers, (2) volunteer training and other organizational needs or requirements, and (3) outcomes, particularly the impact of volunteering on volunteers and the impact of volunteers on EOL care. Despite limited statistical evidence, the available literature on EOL care volunteers clearly indicates that considerable potential benefit can be derived from EOL care volunteers' contributions, with their efforts benefiting dying persons, their families, paid EOL staff, and the volunteers themselves. More specifically, willing volunteers, particularly those with diverse skills and abilities, have the potential to significantly and positively impact EOL care in that they can perform many necessary and extra functions of value. Volunteers often augment and enhance the range of EOL care services provided to terminally ill individuals and their families. Volunteers should also be recognized as increasing the accessibility of EOL care. The role of the volunteer is not without challenge, however, both for the individuals who volunteer and the organizations that must orient them and provide a meaningful role for them. PMID- 16259673 TI - The relationship between income and performance indicators in general practice: a cross-sectional study. AB - OBJECTIVE: To explore the relationship between the income of general practitioners (GPs) and the performance characteristics of their practices. DESIGN: Cross-sectional survey. SETTING: All practices (n = 166) in an inner city health authority, two years before the introduction of the new GP contract in April 2004 were studied. MAIN OUTCOME MEASURES: True income per GP was unavailable to us. Instead, the proxy measure - superannuable pay - was calculated (gross eligible income per GP minus the national average sum for GP expenses). Practice staff funding figures were also obtained. These two financial indicators were compared with practice characteristics and performance indicators. RESULTS: Data were available from 151 out of 166 practices. Based on regression analysis, larger list sizes and higher practice staff budgets predicted 31% of the variation in GP income (standardized beta = 0.66, P < 0.001; beta = 0.19, P = 0.02; respectively). Higher staff budgets were independently associated with better cervical smear and two-year-old vaccination rates (standardized beta = 0.24, P < 0.01; beta = 0.18, P = 0.03; respectively). No association was demonstrated between performance indicators and income. CONCLUSION: Under the previous contract, GPs were able to maximize their income by taking on more patients, whereas achievement of performance targets had very little impact on overall income. The opportunity costs of pursuing higher-quality care might have outweighed the modest financial rewards attached to performance targets. Provided rewards for good-quality care are sufficiently high, the new GP contract is likely to tip the balance in favour of generating earnings by improving the quality of clinical care. To deliver this care, as measured by available performance indicators, our findings imply that a greater investment in practice staff will be needed. PMID- 16259674 TI - Can we use routine data to evaluate organizational change? Lessons from the evaluation of business process re-engineering in a UK teaching hospital. AB - OBJECTIVE: To review and critically evaluate a multidisciplinary independent evaluation of business process re-engineering within a UK hospital using routine data. METHODS: Routine data measures of hospital cost efficiency are used to compare rates of changes at Leicester Royal Infirmary (LRI) with a 'peer group' of 22 teaching hospitals and to ascertain changes attributable to re-engineering. Different adjustment factors are quantified and the robustness of individual measures discussed. In addition to providing context to the quantitative primary research and the qualitative research on change management, service performance and quality indicators not captured in overall efficiency measures are assessed using detailed routine data measures. RESULTS: LRI is one of the most efficient teaching hospitals in England and continued improvement during re-engineering is shown to be at a faster rate than the peer group average, although attribution of changes is difficult. Lack of availability and inconsistency of data for support services meant that many re-engineering initiatives are not quantifiable using routine data measures. CONCLUSIONS: When combining different measures, routine data is a valuable tool in evaluating organizational change initiatives. However, use in future evaluations would require consistent benchmarking of routine data. PMID- 16259683 TI - Whether passenger or patient, organizational safety is the key. PMID- 16259684 TI - A chronic case of mismanagement? PMID- 16259685 TI - Simulation analysis of the consequences of shifting the balance of health care: a system dynamics approach. AB - OBJECTIVES: The shift in the balance of health care, bringing services 'closer to home', is a well-established trend. This study sought to provide insight into the consequences of this trend, in particular the stimulation of demand, by exploring the underlying feedback structure. METHODS: We constructed a simulation model using the system dynamics method, which is specifically designed for the analysis of feedback structure. The model was calibrated to two cases of the shift in cardiac catheterization services in the UK. Data sources included archival data, observations and interviews with senior health care professionals. Key model outputs were the basic trends displayed by waiting lists, average waiting times, cumulative patient referrals, cumulative patient activity and cumulative overall costs. RESULTS: Demand was stimulated in both cases via several different mechanisms. We revealed the roles for clinical guidelines and capacity changes, and the typical responses to imbalances between supply and demand. Our analysis also demonstrated the potential benefits of changing the goals that drive activity by seeking a waiting list goal rather than a waiting time goal. CONCLUSIONS: Appreciating the wider consequences of shifting the balance of care is essential if services are to be improved overall. The underlying feedback mechanisms of both intended and unintended effects need to be understood. Using a systemic approach, more effective policies may be designed through coordinated programmes rather than isolated initiatives, which may have only a limited impact. PMID- 16259686 TI - Demystifying knowledge translation: learning from the community. AB - OBJECTIVES: While there is increasing interest in research related to so-called Knowledge Translation, much of this research is undertaken from the perspective of researchers. The objective of this paper is to explore, through the participatory evaluation of Manitoba's The Need to Know Project, the characteristics of effective knowledge translation initiatives from the perspective of community partners. METHODS: The multi-method evaluation adopted a utilization-focused approach, where stakeholders participated in identifying evaluation questions, and methods were made transparent to participants. Over 100 open-ended, semi-structured interviews were conducted with project stakeholders over the first three years of the project. These interviews explored the perspectives of participants on all aspects of project development. Formal feedback processes allowed further refinement of emerging theory. RESULTS: This research suggests that there has been insufficient emphasis on personal factors in knowledge translation. The themes of 'quality of relationships' and 'trust' connected many different components of knowledge translation, and were essential for collaborative research. Organizational barriers and lack of confidence in researchers present greater challenges to knowledge translation than individual interest or community capacity. The costs of participation in collaborative research for community partners and the benefits for researchers, also require greater attention. CONCLUSIONS: Participation of community partners in The Need to Know Project has provided unique perspectives on knowledge translation theory. It has identified limitations to the common interpretations of knowledge translation principles and highlighted the characteristics of collaborative research initiatives that are of greatest importance to community partners. PMID- 16259687 TI - Urban versus rural populations' views of health care in Scotland. AB - OBJECTIVES: To compare satisfaction with, and expectations of, health care of people in rural and urban areas of Scotland. METHODS: Questions were included in the 2002 Scottish Social Attitudes Survey (SSAS). The Scottish House-hold Survey urban-rural classification was used to categorize locations. A random sample of 2707 people was contacted to participate in a face-to-face interview and a self completion questionnaire survey. SPSS (v.10) was used to analyse the data. Relationships between location category and responses were explored using logistic regression analysis. RESULTS: In all, 1665 (61.5%) interviews were conducted and 1507 (56.0%) respondents returned self-completion questionnaires. Satisfaction with local doctors and hospital services was higher in rural locations. While around 40% of those living in remote areas thought A&E services too distant, this did not rank as a top priority for health service improvement. This could be due to expectations that general practitioners would assist in out of-hours emergencies. Most Scots thought services should be good in rural areas even if this was costly, and that older people should not be discouraged from moving to rural areas because of their likely health care needs. In all, 79% of respondents thought that care should be as good in rural as urban areas. Responses to many questions were independently significantly affected by rural/urban location. CONCLUSIONS: Most Scots want rural health care to continue to be good, but the new UK National Health Service (NHS) general practitioner contract and service redesign will impact on provision. Current high satisfaction, likely to be due to access and expectations about local help, could be affected. This study provides baseline data on attitudes and expectations before potential service redesign, which should be monitored at intervals in future. PMID- 16259688 TI - Adapting the randomized consent (Zelen) design for trials of behavioural interventions for chronic disease: feasibility study. AB - OBJECTIVES: Standard randomized controlled trials of interventions for chronic conditions that involve behavioural change, or that are highly desired by participants, are difficult to undertake because of problems with recruitment and contamination. Alternatives include cluster-randomized trials or pre randomization designs such as the Zelen design. The aim here was to develop a pre randomization design that would overcome ethical and methodological problems associated with the conventional Zelen design, and permit the rigorous evaluation of a complex package of care, involving physical therapy and behavioural changes, for patients with painful patello-femoral osteoarthritis of the knee joint. METHODS: Eligible patients were first consented to a one-year observational study of their arthritis. They were subsequently randomized into intervention and control arms. Those in the intervention arm were then asked if they were willing to participate in a further study involving regular sessions with a physiotherapist. Those in the control arm were not told about this, but were followed up as agreed. RESULTS: Eighty-seven patients consented to the observational study, 43 of whom were subsequently randomized to the intervention arm. All 43 consented to the intervention, although five of these did not receive the full package of care. Assessments were carried out at five months and one year on 82 patients, and concealment was satisfactorily maintained in the majority. CONCLUSIONS: We conclude that this study design could potentially offer an acceptable compromise between the need for scientific rigour and the ethical imperative of fully informed consent in trials that involve behavioural change or interventions that patients might want to obtain. PMID- 16259689 TI - Patient- and ward-level determinants of nursing time in nursing facilities. AB - OBJECTIVES: To explore the patient- and ward-level determinants of wage-adjusted nursing time in long-term care wards for the elderly with a view to improving efficiency of the use of labour resources. METHODS: The wage-adjusted nursing time given to patients was obtained from a time measurement study. Patient characteristics were based on the Minimum Data Set 2.0 of the Resident Assessment Instrument for nursing homes. Ordinary least-squares regression analysis and multilevel modelling were used to disentangle the effect of patient- and ward level factors on nursing time. RESULTS: A significant difference in wage-adjusted nursing time between wards was detected, which was partly explained by characteristics of patients in wards. The combination of patients' physical functioning and cognition and the Resource Utilization Groups RUG-III/22 resource use classification explained 20-25% of patients' nursing time over a 24-hour period. Variables related to the operational environment of the ward did not explain differences in wage-adjusted nursing time once the patient profile of the ward had been controlled for. The results also showed notable unmeasured patient and ward level effects, inefficiency and randomness in the allocation of nursing time. CONCLUSIONS: By improving the allocation and use of labour resources, the substantial variation in nursing time between wards could be diminished. Managers should allocate their staff primarily according to patients' resource needs. PMID- 16259690 TI - Using pharmacy data to identify those with chronic conditions in Emilia Romagna, Italy. AB - BACKGROUND AND OBJECTIVES: Automated pharmacy data have been used to develop a measure of chronic disease status in the general population. The objectives of this project were to refine and apply a model of chronic disease identification using Italian automated pharmacy data; to describe how this model may identify patterns of morbidity in Emilia Romagna, a large Italian region; and to compare estimated prevalence rates using pharmacy data with those available from a 2000 Emilia Romagna disease surveillance study. METHODS: Using the Chronic Disease Score, a list of chronic conditions related to the consumption of drugs under the Italian pharmaceutical dispensing system was created. Clinical review identified medication classes within the Italian National Therapeutic Formulary that were linked to the management of each chronic condition. Algorithms were then tested on pharmaceutical claims data from Emilia Romagna for 2001 to verify the applicability of the classification scheme. RESULTS: Thirty-one chronic condition drug groups (CCDGs) were identified. Applying the model to the pharmacy data, approximately 1.5 million individuals (37.1%) of the population were identified as having one or more of the 31 CCDGs. The 31 CCDGs accounted for 77% (E556 million) of 2001 pharmaceutical expenditures. Cardiovascular diseases, rheumatological conditions, chronic respiratory illness, gastrointestinal diseases and psychiatric diseases were the most frequent chronic conditions. External validation comparing rates of the diseases found through using pharmacy data with those of a 2000 Emilia Romagna disease surveillance study showed similar prevalence of illness. CONCLUSIONS: Using Italian automated pharmacy data, a measure of population-based chronic disease status was developed. Applying the model to pharmaceutical claims from Emilia Romagna 2001, a large proportion of the population was identified as having chronic conditions. Pharmacy data may be a valuable alternative to survey data to assess the extent to which large populations are affected by chronic conditions. PMID- 16259691 TI - A certain ratio? The policy implications of minimum staffing ratios in nursing. AB - The debate about how best to determine nurse staffing levels continues. The conventional wisdom is that determining staffing levels is something best left to local management, taking account of local workload and resources. This 'bottom up' philosophy has now been challenged by the use of a different approach--the use of 'top down'standardized, and mandatory, nurse:patient or nurse:bed ratios. This paper examines the characteristics and early results of the use of staffing ratios in the two health systems where nurse staffing ratios are now mandatory- the states of Victoria (Australia) and California (USA). It then discusses the policy implications of using ratios. The paper identifies the main weaknesses of the use of nurse:patient ratios as being their relative inflexibility and their potential inefficiency, if they are wrongly calibrated. Their strength is their simplicity and their transparency. Their impact will be most pronounced when ratios are mandatory and where they offer a mechanism to improve and then to maintain staffing levels at some pre-determined level. The biggest challenges in their use are calibration (what is 'safe'? or 'minimum'?) and achieving the support of all stake-holders. The paper concludes that nurse:patient ratios are a blunt instrument for achieving employer compliance, where reliance on alternative, voluntary (and often more sophisticated) methods of determining nurse staffing have not been effective. PMID- 16259693 TI - Health as an investment. Catchy idea caught in a time warp? PMID- 16259692 TI - Use of evidence in decision models: an appraisal of health technology assessments in the UK since 1997. AB - OBJECTIVES: To review the sources and quality of evidence used in the development of economic decision models in health technology assessments (HTAs). METHODS: All economic decision models developed as part of the NHS Research and Development HTA Programme between 1997 and 2003 were reviewed. Quality of evidence was assessed using a hierarchy of data sources developed for economic analyses. RESULTS: Decision models are parameterized using diverse sources of evidence (e.g. randomized controlled trials, observational studies, expert opinion). Evidence on the main clinical effect was mostly identified and quality assessed as part of the companion systematic review/meta-analysis of the HTA and therefore reported in a transparent and reproducible way. For the other model inputs (i.e. adverse events, baseline clinical data, resource use and utilities), the search strategies for identifying relevant evidence were rarely made explicit and in a number of reports the sources of specific evidence were unclear due to poor reporting. CONCLUSIONS: A more formal and replicable approach to identification and assessment of quality of model inputs is required to reduce the 'black box' nature of decision models, and lead to less scepticism regarding model outputs. PMID- 16259694 TI - For-profit ownership of facilities would lead to a more efficient health care system. PMID- 16259695 TI - Restructuring acute care hospitals in Newfoundland and Labrador. PMID- 16259696 TI - Acute care restructuring in Newfoundland and Labrador: the history and impact on expenditure. AB - OBJECTIVES: To document the history of regionalization and its effects on the Newfoundland and Labrador acute care health system, and to describe changes in acute care expenditure in the St John's region where hospital redesign, closure and aggregation occurred in relation to other regions not exposed to aggregation. METHODS: Interviews were conducted with senior health officials. Transcripts and other reports were reviewed. Financial data were abstracted from audited general ledger statements received from the Ministry of Health. RESULTS: Regionalization achieved its objectives of hospital aggregation in St John's. The average number of full-time equivalent employees increased slightly by 2% (5304-5416). In some regions, integration of services was delayed because of conflict and resistance to change. There was some disparity between the Provincial Government's objectives for cost control and the CEOs' perceptions of economies of scale. Between 1995/96 and 2002/03, total expenditures for the St John's region and the other five regional hospitals increased by 46% and 54%, respectively; total personal income of the population and government revenues increased by only 18% and 16%, respectively. CONCLUSIONS: Regionalization in Newfoundland and Labrador facilitated aggregation of hospitals, but did not control the number of front line workers and, consequently, total acute care expenditure. Expenditure increased significantly between 1995 and 2002, at a rate which exceeded the increase in government revenues. The government's ability to pay for acute care will not be achieved unless employee costs are controlled or provincial income increases. PMID- 16259697 TI - Regionalization of health services in Newfoundland and Labrador: perceptions of the planning, implementation and consequences of regional governance. AB - OBJECTIVES: To describe the context and key drivers for regionalization of one provincial health care system in Canada; to document the original expectations of regionalization on governance and the extent to which these expectations were met; to identify the perceived successes and weaknesses of the process; and to examine the key issues and concerns that warrant further consideration and action in the future. METHODS: Forty-five CEO/senior administrator or senior health department officials in the period 1993-2001 were invited to participate, of whom 35 were interviewed (67% of senior health officials and 85% of CEOs/senior administrators). RESULTS: For the most part, key informants felt that expectations of reform with respect to reduction in the number of boards and integration of services under each board's mandate did occur. However, ongoing financial restraint, failure to include the full range of health services under the regional board mandate (including physician and pharmaceutical services), uncertainty regarding the level of authority the regional boards had for decision making, and unclear accountability mechanisms between the regional boards and the provincial Ministry of Health limited the extent to which broader expectations related to development of a population health focus, and improved continuity of care for individuals and families was achieved. CONCLUSIONS: Implications for policy-makers were identified in four main areas: alignment between health policy goals and the governance structure; clarification of authority and accountability relationships; clarification of roles and responsibilities among all key actors; and strengthening of mechanisms that support accountability. PMID- 16259698 TI - Attitudes and perceptions of registered nurses during and shortly after acute care restructuring in Newfoundland and Labrador. AB - OBJECTIVES: To monitor changes in registered nurses' perceptions of the impact of seven years of health care restructuring in Newfoundland and Labrador (NL) and to measure the attitudinal and behavioural reactions over four years comparing the St John's region, where hospital aggregation occurred, to other regions of the province. METHODS: Data were collected on acute care nurses' personal characteristics and perceptions of the importance of reform and its impact on workplace conditions and health care quality in 1995, 1999, 2000 and 2002. Nurses' attitudes and intentions were monitored across three time periods (i.e. 1999, 2000 and 2002). RESULTS: Perceived workplace conditions and health care quality, as well as attitudes and behaviours were generally negative. However, there was some improvement over time. The temporal sequence of scores suggests that restructuring had an adverse impact on nurses' attitudes. Few significant regional differences were observed. CONCLUSIONS: Although health services restructuring had an adverse impact on nurses' attitudes, aggregation of hospitals in St John's region was achieved without further deterioration. Provincial wide initiatives are needed to promote more positive work environments and increase the organizational effectiveness. PMID- 16259699 TI - Hospital utilization, efficiency and access to care during and shortly after restructuring acute care in Newfoundland and Labrador. AB - OBJECTIVES: Since the 1990s restructuring, including regionalization and downsizing, has largely been driven by a desire for cost containment. Regionalization, hospital closure and changes in management processes occurred in Newfoundland and Labrador (NL), Canada between 1995 and 2000. The objectives of the current study were: to describe trends in the utilization of acute care hospital services by residents of NL during and shortly after restructuring; to examine trends in the efficiency of utilization of acute care beds in the province during the same time frame; and to compare the trends in St John's with the rest of the province, taking account of confounding events, in an attempt to understand the impact of aggregation of hospitals in this region. METHODS: Hospital discharge and day surgical data were analysed for all facilities in NL from 1995/96 to 2000/01. Analyses were by facility of service and also by region of residence directly standardized to the provincial population for 1996. Efficiency of bed utilization was examined on three occasions by concurrent utilization review using a modified version of the Appropriateness Evaluation Protocol. Trends in the St John's region (where most tertiary services are located and greater aggregation of hospitals occurred) were compared with the rest of the province. RESULTS: Admissions declined by 14% in St John's facilities and by 17% elsewhere. Inpatient days fell by 9% in St John's and by 12% elsewhere. Average length of stay and Resource Intensity Weight changed little, apart from a rise in the final study year, with the largest change in St John's. Standardized hospital admission rates declined by 10% and inpatient days by 5.6% for residents of St John's region, and by 16% and 14% respectively for residents of other regions. There was no change over time in the use of day surgery. Efficiency of acute care bed use improved in 2002 in St John's, but was unchanged in other regions. Use of acute care beds by elderly patients for extended stay, or when an alternate level of care would have been appropriate, was greater in St John's with the disparity persisting over time. Waiting time for continuing care in the St John's region was unchanged comparing 1995/96 and 1999/00. CONCLUSIONS: The degree to which acute care restructuring or financial pressures and constraints imposed at the provincial level contributed to observed utilization trends is unclear. Aggregation of hospitals in the St John's region may have contributed to more efficient use of acute care beds. Restructuring as carried out did not integrate health care sectors, and problems at the acute care/continuing care boundary were not resolved in St John's, where access to continuing care remained difficult. PMID- 16259700 TI - Quality of medical care during and shortly after acute care restructuring in Newfoundland and Labrador. AB - OBJECTIVES: To critically evaluate the quality of hospital medical care at the beginning, during and shortly after regionalization of health boards in Newfoundland and Labrador, and aggregation of hospitals in the St John's region. METHODS: Retrospective chart audits for the years 1995/96, 1998/99 and 2000/01 (at the beginning, during and after restructuring) focused on outcomes in cardiology, respiratory medicine, neurology, nephrology, psychiatry, surgery and women's health programmes. Where possible, quality of care was judged on measurable outcomes in relation to published statements of likely optimal care. Comparisons were made over time within the St John's region, and separately for hospitals in the rest of the province. RESULTS: There was improvement in the use of thrombolytics and secondary measures post-myocardial infarction in both regions. Mortality and appropriateness of initial antibiotic choice for community acquired pneumonia remained stable in both regions, with an improvement in admission appropriateness based on the severity in St John's. Aspects of stroke management (referral and time to see allied health professionals, imaging and discharge home) improved in both regions, while mortality remained stable. There was improvement in fistula rate, quality of dialysis and anaemia management in haemodialysis patients, and improvement in the peritoneal dialysis patient peritonitis rate. Readmission rate for schizophrenia remained unchanged. Stable mortality rates were observed for frequently performed surgical procedures. The post-coronary artery bypass grafting (CABG) morbid event rate improved, although access to CABG was not optimal. CONCLUSIONS: Aggregation of acute care hospitals was feasible without attendant deterioration in patient care, and in some areas care improved. However, access to services continued to be a major problem in all regions. PMID- 16259701 TI - Health care quality from the perspective of health care providers and patients during and shortly after acute care restructuring in Newfoundland and Labrador. AB - OBJECTIVES: To monitor changes in providers' perceptions of health care quality and the importance of health reform, and in patients' satisfaction with services during and two years after restructuring, comparing the region of the province that was restructured (St John's) with those regions in which hospitals were not aggregated. METHODS: The Employee Attitude Survey questionnaire was sent to acute care providers (n = 5353) to assess personal characteristics and perceptions of the impact of reform on workplace conditions, work-related attitudes and turnover intentions. The response rate for 2000 and 2002 was 42% (n = 1222 and 1034, respectively). Only respondents in both surveys (n = 589) were used in the analysis because study results were the same for both the repeat sample and total samples. A Patient Satisfaction Survey questionnaire was administered to patients discharged from acute care facilities in 2000 (n = 1741) and 2002 (n = 704). Response rates were 82.5% and 90.2%, respectively. RESULTS: Most providers felt, at both time periods, that restructuring of the health care system was a positive step, but felt that health care quality was low. In the St John's region, perceptions of quality and standards of care improved over time. Patients were extremely satisfied with the admission process and hospital stay at both time periods in St John's. However, satisfaction declined in 2002 in regions outside St John's. CONCLUSIONS: Aggregation of acute care hospitals is possible without adverse effects on providers' perceptions of health care quality or on patient satisfaction. PMID- 16259702 TI - Health care provider outcomes during and shortly after acute care restructuring in Newfoundland and Labrador. AB - OBJECTIVES: To monitor changes in human resource indicators during six years of restructuring in Newfoundland and Labrador, and to measure providers' perceptions of reform impact and attitudinal and behavioural reactions comparing changes in the St John's region, where hospital aggregation occurred, to other regions. METHODS: Data on human resource indicators from 1995/96 to 2001/02 were obtained and analysed. The Employee Attitude Survey was sent to acute care staff (n = 5353) to assess perceptions of reform impact on workplace conditions, work related attitudes, turnover intentions and personal characteristics. The response rate for 2000 and 2002 was approximately 42% (n = 1222 and 1034, respectively). Only respondents to both surveys (n = 589) were used in the analysis. RESULTS: Increases in average employee and full-time equivalent numbers occurred in the St John's region, despite hospital closure and aggregation. Increases in staff dislocation and turnover were observed, but paid sick hours decreased. Sick leave and overtime costs increased. Although perceived workplace conditions, and attitudes and behaviours were generally negative, there was evidence of improvement over time, especially in St John's. Few significant regional or provider group differences were observed on most study variables. CONCLUSIONS: Aggregation of hospitals in St John's did not lead to a decrease in employee counts, or deterioration in human resource indicators or attitudes. However, province-wide initiatives are needed to promote more positive work environments and increase organizational effectiveness. PMID- 16259703 TI - The Health Care Corporation of St John's: the first five years of regionalization and restructuring. PMID- 16259704 TI - An evaluation of acute care restructuring in Newfoundland and Labrador: conclusions. PMID- 16259706 TI - Special issue to commemorate the life and work of the late Professor Derek Willoughby. PMID- 16259711 TI - Derek Willoughby: an appreciation from a clinical rheumatologist. PMID- 16259713 TI - What if Derek Willoughby were right? Cyclo-oxygenase is an anti-inflammatory enzyme. PMID- 16259714 TI - Prostaglandin F2alpha produced by inducible cyclooxygenase may contribute to the resolution of inflammation. AB - Cyclooxygenase-2 may play a role in resolution of carrageenan-induced pleurisy in rats by generating anti-inflammatory prostanoids. Here, we show exudate prostaglandin F2alpha concentrations rise during resolution of this model. These were reduced by the selective cyclooxygenase-2 inhibitor NS-398, which exacerbated inflammation. Concomitant treatment with NS-398 and the synthetic FP receptor agonist fluprostenol reversed this exacerbation. This suggests prostaglandin F2alpha produced by cyclooxygenase-2 contributes to resolution of this inflammatory reaction. PMID- 16259705 TI - Professor Derek Albert Willoughby (1930-2004). PMID- 16259717 TI - The peroxisome proliferator-activated receptor alpha activator, Wy14,643, is anti inflammatory in vivo. AB - The peroxisome proliferator-activated receptor system is exciting much interest as a novel point of therapeutic intervention in inflammation. Here, the effect of a peroxisome proliferator-activated receptor alpha agonist, [4-chloro-6-(2,3 xylidine)-pyrimidinylthio]acetic acid (Wy14,643), was examined in arachidonic acid-induced murine ear inflammation. 3-[1-(4-Chlorobenzyl)-3-t-butyl-thio-5 isopropylindol-2-yl]-2,2-dimethylpropanoic acid (MK886, a 5-lipoxygenase inhibitor) and indomethacin (a cyclo-oxygenase inhibitor) were used as reference compounds. Wy14,643 dose dependently inhibited ear swelling and polymorphonuclear leukocyte influx, as did MK886, associated with reduced tissue leukotriene B4 but not prostaglandin E2 levels. Unlike MK886, Wy14,643 did not inhibit ex vivo leukotriene B4 production. However, Wy14,643, but not MK886, induced peroxisomal enzyme activity. Indomethacin was less effective, though tissue prostaglandin E2 but not leukotriene B4 levels were reduced. Again, unlike indomethacin, Wy14,643 did not reduce ex vivo prostaglandin E2 production. However, indomethacin did increase peroxisomal enzyme activity but to a lesser extent than Wy14,643. This study demonstrates that peroxisome proliferator-activated receptor alpha activation can inhibit arachidonic acid-induced inflammation in part by enhancing degradation of leukotriene B4. PMID- 16259716 TI - Prostanoids as friends, not foes: further evidence from the interference by cycloxygenase-inhibitory drugs when inducing tolerance to experimental arthritigens in rats. AB - Pharmacologists have generally been prejudiced against prostanoids, uncritically accepting their suppression as desirable therapy, especially for 'quick-fix' analgesia. This myopic perception for a long time ignored (a) the essentiality of prostanoid precursors in nutrition, (b) the physiological protective functions of natural prostaglandins (PGs) (vasculature, stomach, kidney), (c) resolution of inflammation after the expression of COX-2 and (d) increasing therapeutic use of either synthetic PGs (for erectile dysfunction, ophthalmic disorders, inducing parturition, etc) or their natural precursors, e.g., omega3-rich polyunsaturated oils, to treat arthritis. Experimental studies in rats have indicated that prostaglandins (E series) are (i) useful, perhaps auto-regulators of established immunoreactivity and (ii) able to amplify (or even induce) anti-inflammatory activity with other agents. Furthermore, anti-prostanoid therapy (APT) can be arthritigenic!!, interfering with the acquisition of tolerance to some arthritigens. For patients with rheumatoid arthritis this additional side-effect of APT, barely recognised to date, may actually perpetuate their arthritis by impairing prostanoid-mediated remission processes. Hopefully, recent adverse publicity about COX-2 inhibitory drugs might stimulate serious re-assessment of some traditional anti-inflammatory therapies with low APT activity for the management of both acute pain (non-addictive cannabinoids, celery seed, etc.) and chronic inflammation, e.g., Lyprinol (a mussel lipid extract). PMID- 16259719 TI - NMI-1182, a gastro-protective cyclo-oxygenase-inhibiting nitric oxide donor. AB - Non-steroidal anti-inflammatory drugs (NSAIDs) are widely used to treat inflammation and to provide pain relief but suffer from a major liability concerning their propensity to cause gastric damage. As nitric oxide (NO) is known to be gastro-protective we have synthesized a NO-donating prodrug of naproxen named NMI-1182. We evaluated two cyclo-oxygenase (COX)-inhibiting nitric oxide donors (CINODs), NMI-1182 and AZD3582, for their ability to be gastro protective compared to naproxen and for their anti-inflammatory activity. NMI 1182 and AZD3582 were found to produce similar inhibition of COX activity to that produced by naproxen. Both NMI-1182 and AZD3582 produced significantly less gastric lesions after oral administration than naproxen. All three compounds effectively inhibited paw swelling in the rat carrageenan paw edema model. In the carrageenan air pouch model all three compounds significantly reduced PGE2 levels in the pouch exudate but only NMI-1182 and naproxen inhibited leukocyte influx. These data demonstrate that NMI-1182 has comparable anti-inflammatory activity to naproxen but with a much reduced likelihood to cause gastric damage. PMID- 16259718 TI - The role of antioxidants in models of inflammation: emphasis on L-arginine and arachidonic acid metabolism. AB - Inflammatory processes are made up of a multitude of complex cascades. Under physiological conditions these processes aid in tissue repair. However, under pathophysiological environments, such as wound healing and hypoxia-ischaemia (HI), inflammatory mediators become imbalanced, resulting in tissue destruction. This review addresses the changes in reactive oxygen species (ROS), L-arginine and arachidonic acid metabolism in wound healing and HI and subsequent treatments with promising anti-oxidants. Even though these models may appear divergent, anti oxidant treatments are nevertheless still having favourable effects. On the basis of recent findings, it is apparent that protection with anti-oxidants is not solely attributed to scavenging of ROS. In addition, the actions of anti-oxidants must be considered in light of the inflammatory process being assessed. To this end, there does not appear to be any universally applicable single mechanism to explain the actions of anti-oxidants. PMID- 16259720 TI - The inhibition of neutrophil-endothelial cell adhesion by hyaluronan independent of CD44. AB - OBJECTIVE: To study the effect of hyaluronan on cell adhesion and recruitment both in vitro and in vivo, since hyaluronan both inhibits restenosis and is anti inflammatory. When administered to animals undergoing angioplasty the recruitment of cells into the restenotic plaque is inhibited, as well as into inflammatory lesions. The recent discovery that ICAM-1 binds hyaluronan and exhibits the B(X(7))B HA binding motif, led us also to investigate whether cell adhesion could be modulated by hyaluronan. MATERIALS AND METHODS: Human neutrophils were adhered to human umbilical vein (HUVEC) or Ea.hy.926 HUVEC cells stimulated with phorbol myristate acetate (PMA) or tumour necrosis factor (TNFalpha). Neutrophil binding in vivo utilized FMLP-stimulated hamster cheek pouch post-capillary venules. RESULTS: Hyaluronan inhibited human neutrophil adhesion to both PMA and TNFalpha stimulated HUVEC. Ea.hy.926 human immortal HUVECs expressed ICAM-1 in response to TNFalpha and PMA. E-selectin was also upregulated by 6 h with TNFalpha but not significantly with PMA. TNFalpha induced CD44 expression within 4 h, but PMA not significantly up to 6 h. However, specific binding of [125I]hyaluronan to Ea.hy.926 cells was increased by PMA-stimulation at 4 h. Neutrophil adhesion to PMA-stimulated Ea.hy.926 HUVECs was inhibited in a concentration dependent fashion by both anti-ICAM-1 and hyaluronan (1 ng/ml-10 microg/ml) at 4 h. At 1 mg/ml adhesion was stimulated by hyaluronan. Hyaluronan had no effect on neutrophil adhesion to resting Ea.hy.926 cells. Hyaluronan (25 mg/kg, i.v.) inhibited cell adhesion to FMLP-stimulated post capillary venules of the hamster cheek pouch, whilst leaving cell rolling unaffected. CONCLUSIONS: These results show that hyaluronan, at concentrations below those where intra-molecular associations occur, binds selectively to stimulated endothelial cells and inhibits neutrophil adhesion in vitro and in vivo via a mechanism which may involve molecules other than CD44, such as ICAM-1. PMID- 16259721 TI - The modulation of intra-articular inflammation, cartilage matrix and bone loss in mono-articular arthritis induced by heat-killed Mycobacterium tuberculosis. AB - Inflammation models for the assessment of anti-rheumatic drug activity utilize a variety of stimuli and sites. However, the determination of cartilage and bone degradation remains time consuming and problematic. A rapid rat model of Mycobacterium tuberculosis monoarticular arthritis with induces of inflammation, as well as patellar cartilage proteoglycan and bone degradation has been reported. This study characterizes this model with respect to the actions of anti rheumatic drugs. Dexamethasone, cyclosporin and prednisolone inhibited all three parameters. Methotrexate inhibited joint inflammation alone, whilst azathioprine was without effect. Levamisole inhibited cartilage and bone degradation without affecting joint inflammation. NSAIDs were divided in their actions. Naproxen, piroxicam, diclofenac and tiaprofenic acid all inhibited joint inflammation and bone loss, but naproxen and piroxicam both significantly potentiated cartilage proteoglycan loss. This model appears to rely on cellular recruitment at this early stage, the anti-metabolites being ineffective. The modulation of inflammation can result in a protection against cartilage and bone damage in arthritis; however, certain NSAIDs are detrimental to cartilage integrity. The pharmacological manipulation of inflammatory arthritis can therefore dislocate inflammation from its effects on tissue destruction. PMID- 16259723 TI - The kallikrein-kinin system: from mediator of inflammation to modulator of cardioprotection. AB - Kinin is an important mediator of hyperalgesia, inflammatory conditions and asthma. It causes pain, inflammation, increased vascular permeability and vasodilatation. Several kinin antagonists have been developed with the aim of treating these pathologies. Kinin B2 receptor agonists and kallikrein may have clinical utility in the treatment of hypertension, left ventricular hypertrophy, ischemic heart disease, congestive heart failure and diabetes. However, there is a need to know whether there is a safe therapeutic window between potential cardio-protective and pro-inflammatory effects following administration of kinin B2 receptor agonists. PMID- 16259722 TI - Heat-shock proteins and their role in chondrocyte protection, an application for autologous transplantation. AB - Articular cartilage injury presents a unique therapeutic challenge. As cartilage possesses no blood or nerve supply of its own it has a particular susceptibility to early injury and a poor capacity for self-repair. Treatment options are limited and injury can eventually lead to osteoarthritis. Autologous chondrocyte transplantation is an exciting therapeutic development, but despite initial encouraging results, graft failure and formation of fibro- as opposed to hyaline cartilage remain problematic. Bleeding is an inevitable consequence of surgery, and blood-induced cartilage damage is well documented. It is hypothesised here that protecting chondrocytes against blood could significantly improve results. Heat-shock protein induction may confer chondroprotection. The expression of heat shock proteins in human chondrocytes and rat femoral head cartilage following heat shock was analysed by Western blotting, and red-blood-cell-induced chondrocyte death was assessed by cell viability and apoptosis by flow cytometry. We demonstrate that heat-shock induced expression of heat-shock protein 70 (HSP70) (rat and human) and HSP32 (human). Blood and blood products reduced rat cartilage proteoglycan synthesis and human chondrocyte viability, and induced human chondrocyte apoptosis at concentrations considerably lower than those reported previously. The induction of HSP70 in rat cartilage was ineffective in reducing chondrocyte death in the absence or presence of red blood cells or red cell products. Heat shock to human chondrocytes reduced low levels of apoptosis (<20%) and cell death induced by low levels of blood products, but not higher levels. Induction of HSP32 with diacetylrhein appeared to be more effective and may hold greater promise. Blood has potent adverse effects on chondrocytes and the induction and chondroprotective effects of heat-shock proteins could be applied to increase the initial success of implanted chondrocytes improving the outcome of autologous chondrocyte transplantation. PMID- 16259725 TI - Role of cyclooxygenase isoforms in gastric mucosal defense and ulcer healing. AB - The rationale for the development of selective inhibitors of cyclooxygenase-2 (COX-2) was the proposal that this enzyme plays an important role in inflammation but does not contribute to the resistance of the gastrointestinal mucosa against injury. However, studies from several groups have established that both COX-1 and COX-2 have important functions in the maintenance of gastrointestinal mucosal integrity. Thus, in the normal rat stomach lesions only develop when both COX-1 and COX-2 are inhibited. On the other hand, in specific pathophysiological situations the isolated inhibition of either COX-1 or COX-2 without simultaneous suppression of the other COX isoenzyme is ulcerogenic. Furthermore, COX-2 plays an important role in the healing of gastric ulcers and inhibition of COX-2 delays ulcer healing. From these findings the initial concept that only inhibition of COX-1 interferes with gastrointestinal defense has to be re-evaluated. PMID- 16259724 TI - Functional oesophageal epithelial defense against acid. AB - Functional oesophageal epithelial defense, including cell proliferation, restitution, buffers and ion transporters, plays a significant role in maintaining mucosal integrity and enabling rapid repair after injury. Growth factors such as epidermal growth factor (EGF), transforming growth factor-alpha (TGF-alpha), hepatocyte growth factor (HGF) and keratinocyte growth factor (KGF) are associated with oesophageal epithelial proliferation or restitution. Na+/H+ exchanger-1, an ion transporter, regulates intracellular pH and cell volume, and may have roles in cell proliferation, migration and survival. Cytokine, adhesion molecules, cyclooxygenase-2 and free radicals are associated with oesophageal inflammation and breach of the functional epithelial defense. Although the oesophagus does not have strong functional epithelial defense against acid, this defensive mechanisms may be involved in the pathogenesis of non-erosive gastro oesophageal reflux disease. Medical therapy may be developed in future to enhance functional oesophageal epithelial defense. PMID- 16259726 TI - Gastroprotective action of glucocorticoid hormones during NSAID treatment. AB - In this article we present an overview of the results of our studies suggesting that endogenous glucocorticoid hormones play a role as natural defensive factors in maintaining the integrity of the gastric mucosa during treatment with non steroidal anti-inflammatory drugs (NSAIDs). In-domethacin and aspirin at ulcerogenic doses induce a rise in corticosterone, which helps the gastric mucosa to resist the harmful actions of these ulcerogenic agents. The gastroprotective action of glucocorticoids during NSAID treatment may be mediated by multiple actions, including maintenance of glucose homeostasis, mucus production and attenuation of enhanced gastric motility and microvascular permeability. According to our findings, glucocorticoid hormones also participate in the healing processes of NSAID-induced gastric injury. It was demonstrated that there is some cooperative interaction between glucocorticoids and prostaglandins (PGs) in gastroprotection, in a way that a deficiency of one protective factor can lead to an apparently compensatory increase of the other. The gastric mucosa becomes more susceptible to injury during deficiency of both glucocorticoids and PGs. PMID- 16259729 TI - Secretory organelles in ECL cells: effects of pharmacological blockade of the gastrin/CCK2 receptor versus its elimination by gene targeting. AB - Histamine-producing ECL cells are numerous in the stomach. They express gastrin/CCK2 receptors and respond to gastrin by releasing histamine. Ultrastructurally, they display numerous and very characteristic secretory organelles: granules, secretory vesicles and microvesicles. This paper focuses on the impact of the gastrin/CCK2 receptor on the ultrastructure of the ECL cells. The effects of pharmacological blockade of the receptor are compared with the effects of receptor elimination following selective gene targeting. Long-term administration of powerful gastrin/CCK2 receptor antagonists was found to induce hypotrophy of rat stomach ECL cells with reduced number of granules, secretory vesicles and microvesicles. In gastrin/CCK2 receptor knockout mice ECL cells, i.e., histamine-storing cells with the characteristic ultrastructure of ECL cells, had disappeared from the oxyntic mucosa and been replaced by a novel population of endocrine-like cells. These cells harbored granules and microvesicles, but were devoid of histamine and secretory vesicles. We suggest that the gastrin/CCK2 receptor is important for the proper differentiation of the ECL cells and for maintaining their characteristic ultrastructure. PMID- 16259727 TI - Role of central and peripheral ghrelin in the mechanism of gastric mucosal defence. AB - Ghrelin, identified in the gastric mucosa, has been involved in the control of food intake and growth hormone (GH) release, but whether this hormone influences the gastric secretion and gastric mucosal integrity has been little elucidated. We compared the effects of intraperitoneal (i.p.) and intracerebroventricular (i.c.v.) administration of ghrelin on gastric secretion and gastric lesions induced in rats by 75% ethanol or 3.5 h of water immersion and restraint stress (WRS) with or without suppression of nitric oxide (NO)-synthase or functional ablation of afferent sensory nerves by capsaicin. The number and the area of gastric lesions was measured by planimetry, the GBF was assessed by the H2-gas clearance method and blood was withdrawn for the determination of the plasma ghrelin and gastrin levels. In addition, the gastric mucosal expression of mRNA for CGRP, the most potent neuropeptide released from the sensory afferent nerves, was analyzed in rats exposed to WRS with or without ghrelin pre-treatment. Ghrelin (5-80 microg/kg i.p. or 0.6-5 microg/kg i.c.v.) increased gastric acid secretion and attenuated gastric lesions induced by ethanol and WRS. This protective effect was accompanied by a significant rise in the gastric mucosal blood flow (GBF), luminal NO concentration and plasma ghrelin and gastrin levels. Ghrelin-induced protection was abolished by vagotomy and significantly attenuated by L-NNA and deactivation of afferent nerves with neurotoxic dose of capsaicin. The signal for CGRP mRNA was significantly increased in gastric mucosa exposed to WRS as compared to that in the intact gastric mucosa and this was further enhanced in animals treated with ghrelin. We conclude that central and peripheral ghrelin exerts a potent protective action on the stomach of rats exposed to ethanol or WRS, and these effects depend upon vagal activity and hyperemia mediated by the NOS-NO system and CGRP released from sensory afferent nerves. PMID- 16259728 TI - Oxidative stress in Helicobacter pylori-induced gastric cell injury. AB - Oxygen radicals are supposed to be involved in inflammation and cell proliferation. Helicobacter pylori induces decrease in antioxidant defense factors, such as GSH, mucus and constitutive nitric oxide (NO), gastric mucosal injury and inflammation. Inflammation and injury might be caused by oxidant mediated expression of inflammatory cytokine interleukin-8 (IL-8) and inflammatory enzymes such as cyclooxtgenase-2 (COX-2) and inducible nitric oxide synthase (iNOS), which were mediated by oxidant-sensitive transcription factors such as nuclear factor-kappaB (NF-kappaB) and activator protein-1 (AP-1), possibly with mitogen activated protein kinase (MAPK) activation. H. pylori induced alterations in protein expression demonstrate the involvement of oxidative stress in the pathogenesis of H. pylori-induced gastric diseases. The differentially expressed genes and proteins may be useful as prognostic indices for gastric diseases associated with H. pylori infection. In conclusion, oxygen radicals are produced in gastric epithelial cells infected with H. pylori, which may reduce the antioxidant defense mechanism and turn on the expression of inflammatory genes, adhesion molecules and mediators stimulating cell proliferation, as well as defensive molecular chaperones in gastric epithelial cells. PMID- 16259730 TI - Role of the nrf-2 gene in protection and repair of gastric mucosa against oxidative stress. AB - Helicobacter pylori infection, as well as NSAIDs induce oxidative stress on gastric mucosa, thereby causing mucosal damage and retarding mucosal repair. Cells can survive against chronic oxidative stress by enhancing activities of anti-oxidant enzymes, thereby protecting cells from DNA damage. Recent studies have clearly shown that the gene encoding Nrf-2 (NF-E2 p45-related factor-2) plays an important role in the induction of antioxidant enzymes against oxidative stress. In this paper, we will describe the cellular mechanisms by which the nrf 2 gene stimulates anti-oxidant enzyme activities during exposure to oxidative stress. Secondly, we will also mention the beneficial effects of sulforaphane, an isothiocyanate family which is abundantly included in broccoli sprouts, on gastric mucosa. Sulforaphane stimulates nrf-2 gene-dependent anti-oxidant enzyme activities, thereby protecting cells from oxidative injury. Finally, we will state our perspective on the efficacy of sulforaphane in protection and repair of gastric mucosa against oxidative stress during H. pylori infection. PMID- 16259731 TI - Polyamines regulate expression of E-cadherin and play an important role in control of intestinal epithelial barrier function. AB - Epithelial cells line the gastrointestinal mucosa and form an important barrier that protects the subepithelial tissue against a wide array of noxious substances, allergens, viruses and luminal microbial pathogens. Restoration of mucosal integrity following injury and various environmental stresses requires epithelial cell decisions that regulate signaling networks controlling gene expression, survival, migration and proliferation. Recently, it has been shown that polyamines play an important role in the regulation of cell-cell interactions and are critical for maintenance of intestinal epithelial integrity. Both the function of polyamines in expression of adherens junction proteins and their possible mechanisms, especially in implication of intracellular Ca2+ and c Myc transcription factor, are the subject of this review article. PMID- 16259732 TI - Intestinal dysmotility in inflammatory bowel disease: mechanisms of the reduced activity of smooth muscle contraction. AB - Inflammation suppresses intestinal motility, which secondarily induces abnormal growth of intestinal flora. Disturbance of this flora plays a role in the pathogenesis of mucosal inflammation, which in turn aggravates the intestinal dysmotility. Therefore, it is important to know the mechanism of alteration in motor function in the inflamed intestine. Recent studies have shown molecular mechanisms responsible for the motility disorder in the inflamed gut. These include an increase in the activity of myosin light-chain phosphatase and an alteration of ion channel activity in smooth muscle cells. PMID- 16259734 TI - Co-localization of TRPV1-expressing nerve fibers with calcitonin-gene-related peptide and substance P in fundus of rat stomach. AB - The localization of vanilloid receptor TRPV1 was studied in rat gastric fundus by an immunohistochemical technique. Numerous TRPV1-immunoreactive nerve fibers were found around arterioles in the submucosal layer. A large number of the nerve fibers were also seen in the smooth muscle layer and in the myenteric nerve plexus, but the cell bodies could not be found. TRPV1 nerve fibers within the circular muscle layers were running parallel to the muscle fibers. Virtually all TRPV1 axons were immunoreactive for calcitonin-gene-related peptide (CGRP), with particularly extensive double labeling seen in axons of the submucosa around blood vessels. TRPV1 nerve fibers containing substance P were found running in longitudinal muscle and circular muscle. The TRPV1 axons seem to be predominantly extrinsic and contain CGRP and substance P in gastric fundus. TRPV1 neurons are thought to be sensory afferent neurons that operate to maintain gastric motility and blood flow. PMID- 16259735 TI - Participation of vanilloid/capsaicin receptors, calcitonin-gene-related peptide and substance P in gastric protection of omeprazole and omeprazole-like compounds. AB - The effects of omeprazole and different omeprazole-like compounds, associated with anti-ischaemic, antioxidant and poly(adenosine-diphosphate-ribose) polymerase (PARP) inhibitory properties, on the gastric acid secretion (4 h pylorus-ligated) and indomethacin-induced gastric mucosal damage connected with the specific immunohistochemical distribution of TRPV1, CRGP and SP during the effects of these compounds, were studied. The observations were carried out in CFY-strain rats (180-210 g), according to the standard methods and the above mentioned parameters were studied in these experimental circumstances without and with application of different compounds. We found that: (1) all of the compounds dose-dependently inhibited the gastric acid secretion and mucosal damage; (2) the expression of TRPV1 receptor, CGRP and SP decreased significantly in both pylorus ligated and indomethacin-treated animals and (3) the expression of TRPV1 and CGRP was reduced. Meanwhile, no change was obtained in SP expression during the gastric mucosal protection produced by omeprazole and omeprazole-like compounds. The conclusions were that (1) a functional overlap exists between the capsaicin sensitive afferent and efferent vagal nerve during omeprazole effects; (2) chemical modification of omeprazole molecule offers a new pathway to obtain a new drug for the introduction in the clinical practice. PMID- 16259733 TI - AP-1 and colorectal cancer. AB - Activator protein-1 (AP-1) is a transcription factor that consists of either a Jun-Jun homodimer or a Jun-Fos heterodimer. AP-1 regulates the expression of multiple genes essential for cell proliferation, differentiation and apoptosis. Numerous reports suggest that AP-1 plays an important role in various human diseases. Among them, the roles relating to human cancers have been strongly suggested for a long time. In human cancers, colorectal cancer is still a leading cause of morbidity and mortality in the world. Since there are some reports about the role of AP-1 in colorectal cancer response to a number of stimuli, such as cytokines and growth factors, and oncogenictransformation, therapeutic inhibition of AP-1 activity has attracted considerable interest. Here, we demonstrate the biological properties of AP-1 and its role in colorectal cancer, and discuss a possibility of an AP-1 inhibitor, an adenovirus dominant-negative mutant of c Jun, as a therapeutic agent for gene therapy. PMID- 16259736 TI - Immunohistochemical distribution of vanilloid receptor, calcitonin-gene related peptide and substance P in gastrointestinal mucosa of patients with different gastrointestinal disorders. AB - The immunohistochemical distribution of capsaicin/vanilloid (transient receptor potential vanilloid 1, TRPV1) receptors and neuropeptides (CGRP, SP) was studied in the gastrointestinal mucosal biopsies of patients with gastritis, erosions, ulcers, polyps, adenocarcinoma, chronic inflammatory bowel diseases, polyps without and with hyperplasia, dysplasia and adenocarcinoma in colon. The studies were carried out in 127 patients and 30 people with only functional dyspepsia (without any histological alteration). The results were: (1) the positivity of TRPV1 receptor and CGRP was detected, and weak participation of SP was detected in patients with different gastric diseases; (2) the presence of TRPV1, CGRP and SP could be detected in chronic inflammation of bowel disease; (3) SP could not detected in patients with colon polyps, dysplasia and adenocarcinoma; (4) the presence of TRPV1 and CGRP was proved in colon dysplasia and adenocarcinoma. We conclude that (1) the immunohistochemical distribution of TRPV1, CGRP and SP differs in gastrointestinal diseases of the upper and lower tract, and (2) the participation of TRPV1, CGRP and SP differs significantly in these different gastrointestinal diseases. PMID- 16259738 TI - Acid secretion in urinary bladder of rats subjected to gastrocystoplasty. AB - Urinary bladder augmentation with a segment of the stomach, i.e., gastrocystoplasty, has been used to improve capacity and compliance in patients with bladder dysfunction. In the present study, rats were subjected to gastrocystoplasty (using the oxyntic segment) with or without fundectomy (removal of the oxyntic part of stomach), and the acid secretion in the augmented bladder was measured. In freely fed rats, the pH values were neutral and not significantly decreased in the rats subjected to gastrocystoplasty with or without fundectomy compared to controls (no operation or sham operation). In response to food intake after being fasted, the rats subjected to gastocystoplasty + fundectomy produced significant amounts of acid. Immunohistochemical examination revealed that the ECL cells and parietal cells seemed to be normal in rats with gastrocystoplasty alone, and that micronodules of ECL appeared to develop in rats with gastrocystoplasty + fundectomy. We suggest that the rats subjected to gastrocystoplasty + fundectomy are capable of producing acid secretion in the bladder, probably due to the secretagogue and trophic effects of gastrin on the ECL cells in the segment of the oxyntic mucosal segment of the bladder. PMID- 16259739 TI - Genetic dissection of the signaling pathways that control gastric acid secretion. AB - Gastric acid secretion is regulated by endocrine, paracrine and neurocrine signals via at least three pathways, the gastrin-histamine pathway, the CCK somatostatin pathway and the neural pathway. Genetically-engineered mice, subjected to targeted gene disruption (i.e., knockout mice), have been used to dissect the signaling pathways that are responsible for the complexity of the regulation of acid secretion in vivo. Both gastrin knockout and gastrin/CCK2 receptor knockout mice displayed greatly impaired acid secretion, presumably because of the loss of the gastrin-histamine pathway. Gastrin/CCK double-knockout mice had a relatively high percentage of active parietal cells with a maintained ability to respond with copious acid secretion to pylorus ligation-evoked vagal stimulation and to a histamine challenge. The low acid secretion in gastrin knockout mice and gastrin/CCK2 receptor knockout mice and the restoration of acid secretion in gastrin/CCK double-knockout mice suggest that CCK plays an important role as inhibitor of the parietal cells via the CCK-somatostatin pathway by stimulating the CCK1 receptor of the D cell. In the absence of both the gastrin histamine and the CCK-somatostatin pathway (as in gastrin/CCK2 receptor double knockout mice), the control of acid secretion is probably taken over by neural pathways, explaining the high acid output. The observations illustrate the complexity and plasticity of the acid regulatory mechanisms. It seems that one pathway may be suppressed or allowed to dominate over the others depending on the circumstances. PMID- 16259740 TI - Development of intestinal, but not gastric damage caused by a low dose of indomethacin in the presence of rofecoxib. AB - The ulcerogenic effect of rofecoxib, a selective cyclooxygenase-2 (COX-2) inhibitor, on the gastrointestinal mucosa was investigated in the presence of a low dose of indomethacin. Indomethacin at 3 mg/kg did not cause any damage in both the stomach and small intestine, despite inhibiting prostaglandin (PG) production. Rofecoxib had no effect on PG production and did not cause any damage in these tissues. In the presence of indomethacin, however, rofecoxib provoked damage in the small intestine but not the stomach. Indomethacin at 3 mg/kg induced hypermotility and COX-2 expression in the intestine but not in the stomach, both in an atropine-sensitive manner. These results suggest that a low dose of indomethacin produces damage in the small intestine but not in the stomach when administered together with rofecoxib. The PG deficiency caused by a low dose of indomethacin produces hypermotility and COX-2 expression in the small intestine, and results in damage when COX-2 is inhibited. It is assumed that the hypermotility response is a key event in the expression of COX-2 and thereby important in the development of mucosal damage in the gastrointestinal tract. PMID- 16259737 TI - Gastric HCO3- secretion induced by mucosal acidification: different mechanisms depending on acid concentration. AB - We compared the HCO3- secretory responses induced by mucosal acidification at different HCl concentrations (100 and 200 mM HCl) in the rat stomach. Under urethane anesthesia, the stomach was mounted on an ex vivo chamber and perfused with saline under inhibition of acid secretion by omeprazole (60 mg/kg, i.p.). TheHCO3- secretion was measured at pH 7.0 using a pH-stat method and by adding 2 mM HCl. The acidification was performed by exposure of the mucosa to 100 mMor 200 mM HCl for 10 min. The secretion of HCO3- was increased by acidification of the mucosa at both 100 and 200 mM of HCl, and the maximal HCO3- response was 1.5 times greater at the latter concentration. The HCO3- responses induced by 100 and 200 mM HCl were both totally inhibited by prior administration of indomethacin, an inhibitor of prostaglandin (PG) production. The HCO3- stimulatory effect of 200 mM HCl was also significantly attenuated by pre-treatment with N(G)-nitro L arginine methyl ester (L-NAME), the inhibitor of nitric oxide (NO) synthase, as well as chemical ablation of capsaicin-sensitive afferent neurons, whereas that of 100 mM HCl was affected by neither of these treatments. We conclude that the mucosal acidification stimulates gastric HCO3- secretion in different mechanisms, depending on the concentration of acid; the response caused by 100 mM HCl is mediated only by PGs, while that caused by 200 mM HCl is mediated by both capsaicin-sensitive afferent neurons and NO, in addition to PGs. PMID- 16259741 TI - Gastroprotective action of glucocorticoid hormones in rats with desensitization of capsaicin-sensitive sensory neurons. AB - The ability of glucocorticoid hormones to protect gastric mucosa during desensitization of capsaicin-sensitive afferent neurons has been investigated in rats. Functional ablation of the afferent neurons was performed by pre-treatment with neurotoxic doses of capsaicin (100 mg/kg s.c.). After 1 week of recovery, capsaicin-desensitized, as well as control rats were adrenalectomized or sham operated. Seven days later, indomethacin at an ulcerogenic dose (35 mg/kg s.c.) was given to each group of rats. One half of adrenalectomized capsaicin-pre treated rats were injected by corticosterone for replacement (4 mg/kg s.c., 15 min before indomethacin). Gastric lesions, plasma corticosterone and blood glucose levels were estimated 4 h after indomethacin administration. Indomethacin caused gastric erosions that were aggravated by adrenalectomy or desensitization of capsaicin-sensitive afferent neurons approximately with the same extension. Combination of adrenalectomy with the sensory desensitization profoundly potentiated the effect of sensory desensitization alone on indomethacin-induced gastric erosions: the mean gastric erosion area was increased approximately 10 fold. Corticosterone replacement completely prevented this profound effect of adrenalectomy. The results suggest a pivotal role of glucocorticoid hormones in the maintenance of gastric mucosal integrity in the case of impaired gastroprotective mechanisms provided by PGs and capsaicin-sensitive sensory neurons. PMID- 16259742 TI - Gastric bypass surgery does not increase susceptibility to Helicobacter pylori infection in the stomach of rat or mouse. AB - Gastric bypass is a clinical option for obesity surgery. An increased susceptibility to Helicobacter pylori infection in the bypassed stomach has been speculated. The aim of the present study was to examine the susceptibility of the bypassed stomach to H. pylori infection in rats and mice. Adult Sprague-Dawley and Wistar rats and NMRI mice were subjected to either gastric bypass or laparotomy only as control. The animals were inoculated with the CagA- and VacA- positive H. pylori strain 67/21 (not mouse-adapted) in the first experiment and with 9 additional isolates in the second, by injection into the bypassed stomach or the control stomach during surgery. The stomach of each animal was collected for H. pylori culture 2-3 weeks later. While all the rats were H. pylori negative, 54% of gastric bypassed mice and 75% of controls were positive (P = 0.4). We conclude that susceptibility to H. pylori infection in the stomach is not increased by gastric bypass surgery. PMID- 16259745 TI - A novel gastric lesion model induced in rats by partial gastric vascular ligation. AB - Previous studies have suggested that histamine treatment after gastric vascular ligation induces mucosal damage in the rat stomach. Although ligation of left gastric artery and vein (L-AV) alone did not cause any damage in the stomach within 4 h, but provoked mild lesions due to ischaemia 24 h later. In the present study we demonstrated a new model of gastric lesions induced by L-AV ligation and examined the effects of various anti-ulcer drugs on this lesion model. The gastric lesions induced by L-AV ligation occurred at the corpus and antrum, especially at the corpus-antrum border, when examined 24 h later, and the severity of damage reached maximum 3 days after L-AV ligation. Repeated treatment with omeprazole or sucralfate for 3 days significantly prevented the development of gastric lesions induced by L-AV ligation, in whole mucosa, including the antrum. By contrast, famotidine given for 3 days showed a significant protection against total lesions in the whole mucosa, but had no effect on the antral lesions. Both omeprazole and famotidine dose-dependently decreased gastric acid output while sucralfate raised the intraluminal pH due to the acid-neutralizing action. These results suggest that the pathogenesis of gastric lesions induced by L-AV ligation differs depending on the region, the corpus and the antrum, and the lesions occurred in the latter area seem to be resistant to acid suppression. It is assumed that this new model of gastric lesions is useful for screening the drugs that affect gastric mucosal defense rather than acid secretion. PMID- 16259744 TI - Role of gastric mucosal ascorbic acid in gastric mucosal lesion development in rats with water immersion restraint stress. AB - We examined the role of gastric mucosal ascorbic acid (AA) in gastric mucosal lesion development in rats with water immersion restraint stress (WIRS). When fasted rats were subjected to WIRS for 1, 3 or 6 h, gastric mucosal lesions developed at 3 and 6 h. Gastric mucosal AA concentration decreased at 3 and 6 h after the onset of WIRS, while gastric mucosal non-protein SH concentration decreased at 1, 3, and 6 h and gastric mucosal vitamin E concentration decreased at 6 h. Gastric mucosal lipid peroxide concentration and myeloperoxidase activity increased at 3 and 6 h of WIRS. Pre-administration of AA (250 mg/kg) prevented gastric mucosal development with attenuation of the decreased gastric mucosal AA, non-protein SH and vitamin E concentrations, and the increased gastric mucosal lipid peroxide concentration and myeloperoxidase activity. These results suggest that gastric mucosal AA plays an important role in WIRS-induced gastric mucosal lesion development. PMID- 16259746 TI - Irritative action of alcoholic beverages in rat stomachs: a comparative study with ethanol. AB - The mucosal irritative action of alcoholic beverages such as white wine, Japanese sake and whisky was examined in rat stomachs in vivo and in vitro, in comparison with ethanol. The concentration of ethanol in these alcoholic beverages was 15%. Mucosal application of ethanol (15%) and whisky in the chambered stomach caused a decrease in gastric potential difference (PD), while that of Japanese sake and white wine caused a slight increase but not decrease in PD. Likewise, both ethanol and whisky markedly reduced the cell viability of RGM1 cells after 5 min incubation, whereas neither Japanese sake nor white wine had any effect. In addition, supplementation of glucose, one of the non-alcoholic ingredients of white wine and Japanese sake, antagonized a reduction in both PD and cell viability caused by ethanol. These results suggest that the mucosal irritative action of Japanese sake and white wine is much less than that of ethanol or whisky and that these properties may be, at least partly, due to the glucose contained in these alcoholic beverages. PMID- 16259743 TI - Modulation of gastric hemorrhage and ulceration by oxidative stress and histamine release in Salmonella typhimurium-infected rats. AB - Infection with Salmonella typhimurium can produce multiple organ dysfunctions. However, document concerning with gastric hemorrhagic ulcers occur in this infectious disease is lacking. The aim was to study modulation of gastric hemorrhagic ulcer by oxidative stress and mast cell histamine in S. typhimurium infected rats. Additionally, the protective effects of drugs, such as ofloxacin, lysozyme chloride, ketotifen, ranitidine, and several antioxidants, including exogenous glutathione (GSH), allopurinol and dimethylsulfoxide (DMSO) were evaluated. Male Wistar rats were injected intrajejunally with a live culture of S. typhimurium (1 x 10(10) colony-forming units/rat) and followed by deprivation of food for 36 h. Age-matched control rats received sterilized vehicle only. Rat stomachs were irrigated for 3 h with either normal saline or a simulated gastric juice containing 100 mM HCl, 17.4 mM pepsin and 54 mM NaCl. S. typhimurium caused aggravation of offensive factors, including enhancing gastric acid back diffusion, mucosal lipid peroxide generation, histamine release, microvascular permeability and hemorrhagic ulcer, as well as an attenuation of defensive substances, such as mucosal GSH and mucus level. Intragastric irrigation of gastric juice caused further aggravation of these gastric biochemical parameters. This exacerbation of ulcerogenic factors was abolished by pretreatment of ofloxacin and lysozyme chloride. Antioxidants, such as reduced GSH, allopurinol and DMSO also produced significant (P < 0.05) amelioration of gastric damage in S. typhimurium infected rats. In conclusion, gastric oxidative stress and histamine play pivotal roles in the formation of hemorrhagic ulcers that were effectively ameliorated by ofloxacin, lysozyme chloride, ketotifen, ranitidine, diamine oxidase and various antioxidants in S. typhimurium-infected rats. PMID- 16259747 TI - Histamine regulates growth of malignant melanoma implants via H2 receptors in mice. AB - The present study examined the effect of histamine H2-receptor antagonists and exogenous histamine on growth of malignant melanoma implant in mice. Drugs were administered to B16BL6 malignant-melanoma-implanted syngeneic mice, and the tumor volume was measured throughout the experiments. Cell proliferation was assessed by MTT assay and mRNA expression was determined by RT-PCR. Both roxatidine and cimetidine significantly suppressed growth of B16BL6 implant compared with vehicle. On the other hand, systemically administered histamine significantly stimulated growth of B16BL6 implants. In addition, the histamine-stimulated B16BL6 implant growth was markedly suppressed by co-administration of cimetidine in a dose-dependent manner. H2-receptor antagonists, however, failed to affect in vitro proliferation of B16BL6 cells. H2-receptor mRNA was detected in B16BL6 implants but not in the cell line. These results indicated that both endogenous and exogenous histamine have ability to stimulate growth of malignant melanoma implants via H2 receptors expressed in host cells. PMID- 16259751 TI - Drug delivery to tumours: recent strategies. AB - Despite several advancements in chemotherapy, the real therapy of cancer still remains a challenge. The development of new anti-cancer drugs for the treatment of cancer has not kept pace with the progress in cancer therapy, because of the nonspecific drug distribution resulting in low tumour concentrations and systemic toxicity. The main hindrance for the distribution of anti-cancer agents to the tumour site is the highly disorganized tumour vasculature, high blood viscosity in the tumour, and high interstitial pressure within the tumour tissue. Recently, several approaches such as drug modifications and development of new carrier systems for anti-cancer agents have been attempted to enhance their tumour reach. Approaches such as drug delivery through enhanced permeability and retention (EPR) effect have resulted in a significant improvement in concentration in tumours, while approaches such as drug-carrier implants and microparticles have resulted in improvement in local chemotherapy of cancer. This review discusses different strategies employed for the delivery of anti-cancer agents to tumours, such as through EPR effect, local chemotherapeutic approaches using drug delivery systems, and special strategies such as receptor-mediated delivery, pH-based carriers, application of ultrasound and delivery to resistant tumour cells and brain using nanoparticles. PMID- 16259749 TI - Lansoprazole binding to the neutrophils in dextran sulfate sodium-induced rat colitis. AB - To clarify the effector sites of lansoprazole in the colonic mucosa during the formation of colitis, dextran sulfate sodium-induced colitis was induced by the oral administration of 3% aqueous solution for 3 and 7 days. The effector sites of [3H]lansoprazole were examined by the intra-aortic infusion of the radiolabelled compound and the autoradiographic tracing of water-soluble compounds. As a result, the [3H]lansoprazole binding in the control rat colon was negligible, while in dextran sulfate sodium-treated rats specific binding sites of [3H]lansoprazole were recognized in the cytoplasm of the mesenchymal cells, and most of them coincided with polymorphonuclear leucocytes and macrophages. PMID- 16259750 TI - Agonist of peroxisome proliferator-activated receptor gamma (PPAR-gamma): a new compound with potent gastroprotective and ulcer healing properties. AB - Pioglitazone, a specific ligand for peroxisome proliferator-activated receptor gamma (PPAR-gamma), was recently implicated in the control of inflammatory processes and in the modulation of the expression of various cytokines such as tumor necrosis factor alpha (TNF-alpha), but its role in the mechanism of gastric mucosal integrity has not been studied extensively. This study was designed to determine the effect of pioglitazone on gastric mucosal lesions induced in rats by topical application of 100% ethanol and by 3.5 h of water immersion and restraint stress (WRS) with or without pretreatment with indomethacin (5 mg/kg i.p.) to inhibit cyclooxygenase-1 (COX-1) and COX-2 enzyme activities and L-NNA (20 mg/kg i.p.) to suppress nitric oxide (NO)-synthase. In addition, the effect of pioglitazone on ulcer healing in rats with chronic acetic acid ulcers (ulcer area 28 mm2) was determined. Rats were killed 1 h and 3.5 h after ethanol administration or WRS exposure or at day 9 upon ulcer induction, and the number and area of gastric lesions were measured by planimetry, the gastric blood flow (GBF) was determined by H2-gas clearance technique and the mucosal PGE2 generation and gene expression and plasma concentration of TNF-alpha and IL-1beta were also evaluated. Pre-treatment with pioglitazone dose-dependently attenuated gastric lesions induced by 100% ethanol and WRS; the dose reducing these lesions by 50% (ID50) being 10 mg/kg and 7 mg/kg, respectively. The protective effect of pioglitazone was accompanied by the significant rise in the GBF, an increase in PGE2 generation and the significant fall in the plasma TNF-alpha and IL-1beta levels. Strong signals for IL-1beta- and TNF-alpha mRNA were recorded in gastric mucosa exposed to ethanol or WRS, and these effects were significantly decreased by pioglitazone. Indomethacin which suppressed PG generation by about 90%, while augmenting WRS damage, and L-NNA, that suppressed NO-synthase activity, significantly attenuated the protective and hyperaemic activity of this PPAR gamma ligand. In the chronic study, pioglitazone significantly reduced the area of gastric ulcers on day 9 and significantly raised the GBF at the ulcer margin. The acceleration of ulcer healing by PPAR-gamma ligand was accompanied by a significant increase in the expression of PECAM-1 protein, a marker of angiogenesis. We conclude that (1) pioglitazone exerts a potent gastroprotective and hyperaemic actions on the stomach involving endogenous PG and NO and attenuation of the expression and release of proinflammatory cytokines TNF-alpha and IL-1beta, and (2) PPAR-gamma ligand accelerates ulcer healing, possibly due to the enhancement in angiogenesis at ulcer margin. PMID- 16259748 TI - Different effects of dexamethasone and the nitric oxide synthase inhibitor L-NAME on caerulein-induced rat acute pancreatitis, depending on the severity. AB - Effects of dexamethasone and N(G)-nitro-L-arginine methyl ester (L-NAME), the nitric oxide (NO) synthase inhibitor, on caerulein-induced acute pancreatitis were examined in rats. Acute pancreatitis was induced by caerulein (20 mug/kg, s.c.) given repeatedly 2 or 4 times every hour, and serum amylase levels, pancreas weight and myeloperoxidase (MPO) activity were measured 6 h after the first injection of caerulein. Dexamethasone (3 mg/kg) and L-NAME (30 mg/kg) were administered p.o. 30 min before the first injection of caerulein. Caerulein caused moderate or severe pancreatitis, depending on the times of injections, resulting in different degrees of increase in serum amylase levels and pancreas weight, and the marked elevation of MPO activity was observed only after injections of caerulein given 4 times per hour. Both dexamethasone and L-NAME suppressed the severity of pancreatits, yet the effect of L-NAME as compared with dexamethasone was more potent against mild pancreatitis but less potent against severe pancreatitis. These results suggest that caerulein-induced acute pancreatitis shows different responsiveness to L-NAME and dexamethasone, depending on the severity; the former is more effective against pancreatitis with less inflammation, while the latter is more effective against pancreatitis with severe inflammation. It is assumed that endogenous NO may be involved in oedema formation as the early event in the development of acute pancreatitis. PMID- 16259752 TI - Hydrodynamic simulation (computational fluid dynamics) of asymmetrically positioned tablets in the paddle dissolution apparatus: impact on dissolution rate and variability. AB - The aim of this work was to investigate the dissolution rate from both the curved and planar surfaces of cylindrical compacts of benzoic acid, which were placed centrally and non-centrally at the base of the vessel of the paddle dissolution apparatus. The effect of fixing the compacts to a particular position on the variability of dissolution results was also examined. In addition, computational fluid dynamics (CFD) was used to simulate fluid flow around compacts in the different positions in the vessel, and the relationship between the local hydrodynamics in the region of the compacts and the dissolution rate determined. The dissolution rate was found to increase from the centre position to the off centre positions for each surface examined. There was a corresponding increase in maximum fluid velocities calculated from the CFD fluid flow simulations at a fixed distance from the compact. There was less variability in dissolution from compacts fixed to any of the positions compared with those that were not fixed. Fluid flow around compacts in different positions could be successfully modelled, and hydrodynamic variability examined, using CFD. The effect of asymmetric fluid flow was evident visually from the change in shape of the eroded compacts. PMID- 16259753 TI - Characterization of crosslinking effects on the physicochemical and drug diffusional properties of cationic hydrogels designed as bioactive urological biomaterials. AB - This study examined the effects of concentration and type of crosslinker (tetraethyleneglycol diacrylate, TEGDA; diethyleneglycol dimethacrylate, DEGDMA; and polyethyleneglycol dimethacrylate, PEGDMA) on the mechanical and drug diffusional properties of hydrogels that had been selected as candidate coatings for bioactive medical devices. Hydrogels (dimethylaminoethylmethacrylate-co vinylpyrrolidone; 1:1) were prepared by free radical polymerization and characterized using tensile analysis, dynamic contact angle analysis and analysis of swelling at pH 6.0. The release of fusidic acid and chlorhexidine was evaluated using buffered medium at pH 6.0 and, in addition, using dissolution medium that had been buffered to pH 9 in the presence and absence of elevated concentrations of calcium, representative of urinary encrustation. Crosslinker concentration, but not type, affected the advancing and receding contact angles. Conversely, both crosslinker type and concentration affected the mechanical and swelling properties of the hydrogels. Maximum swelling and elongation at break were associated with the PEGDMA-crosslinked hydrogels whereas TEGDA-crosslinked hydrogels exhibited the maximum ultimate tensile strength and Young's modulus. Drug release from all systems occurred by diffusion. The mass of chlorhexidine and fusidic acid released was dependent on crosslinker type and concentration, with hydrogels crosslinked with PEGDMA offering the greatest mass of drug released at each sampling period. The mass of fusidic acid but not chlorhexidine released at pH 9.0 in a calcium augmented medium was lower than that released in the same medium devoid of elevated calcium, due to the formation of the poorly soluble calcium salt. In conclusion, this study has uniquely examined the effects of crosslinker type and concentration on physicochemical and drug release properties essential to the clinical and non-clinical performance of bioactive hydrogels for medical device application. PMID- 16259754 TI - Effect of the molecular weight of poly(ethylene glycol) used as emulsifier on alpha-chymotrypsin stability upon encapsulation in PLGA microspheres. AB - Poly(ethylene glycol) (PEG) was used as emulsifier to prepare alpha-chymotrypsin loaded poly(lactic-co-glycolic) acid (PLGA) microspheres by a solid-in-oil-in water (s/o/w) technique. The effect of the molecular weight of PEG on protein stability was assessed by the determination of the amount of insoluble aggregates, the activity loss and the magnitude of structural perturbations. In addition, the effect of the molecular weight of PEG on the encapsulation efficiency, microsphere characteristics and release kinetics was investigated. X ray photoelectron spectroscopy (XPS) was employed to characterize the surface chemistry of the microspheres. Microspheres were prepared using PEG with molecular weight of 6,000, 8,000, 10,000, 12,000 and 20,000. The results indicate that PEG 20,000 was the most effective emulsifier when producing alpha chymotrypsin-loaded microspheres with respect to protein stability. The aggregate formation was decreased from 18% to 3%; the protein inactivation and the encapsulation-induced structural perturbations were largely prevented. XPS confirmed that PEG was largely located on the surface of microspheres. The molecular weight of PEG affected the microspheres' characteristics and release kinetics. Microspheres prepared with PEG 20,000 showed improved encapsulation efficiency (80%) and a continuous release (for 50 days) with the lowest amount of initial release. It is demonstrated that the selection of the optimum molecular weight of PEG when used as emulsifier in the preparation of microspheres is a critical factor in the development of sustained-release formulations for the delivery of proteins. PMID- 16259755 TI - Differential molecular interactions between the crystalline and the amorphous phases of celecoxib. AB - We have investigated the differences in molecular interactions between the crystalline (ordered) and amorphous (disordered) phase of a poorly soluble drug, celecoxib. Molecular interactions in the crystalline phase were investigated with the help of Mercury software, using single crystal X-ray diffractometric data for celecoxib. A simulated annealing molecular dynamics approach was used for the assessment of altered molecular interactions in the amorphous phase. Crystalline celecoxib was found to contain an ordered network of H-bonding between all its electron donors (-S=O group, 2-N of pyrazole ring and -C-F) and the acceptor (-N H). Amorphous celecoxib retained all these interactions in its disordered molecular arrangement, with a relatively stronger H-bonding between the interacting groups, as compared with crystalline celecoxib. However, these inter molecular interactions differed in strength in the two solid-state forms. The altered configurations of the molecular arrangement in the two phases were supported by the shifts observed in the Fourier-transform infra-red vibrational spectra of respective states. These interactions could have strong implications on devitrification kinetics of amorphous celecoxib, and could further guide the choice of stabilizers for the amorphous form. PMID- 16259756 TI - In-vitro cytotoxicity, in-vivo biodistribution and anti-tumour effect of PEGylated liposomal topotecan. AB - In attempt to increase the accumulation of topotecan in tumours and improve its anti-cancer activity, PEGylated liposome (H-PEG) containing topotecan was prepared. The in-vitro cytotoxicity, in-vivo biodistribution pattern and anti tumour effect of H-PEG were studied systemically. Compared with free topotecan or conventional liposome (H-Lip), H-PEG improved the cytotoxic effect of topotecan against human ovarian carcinoma A2780 and human colon carcinoma HCT-8 cells. The IC50 value (concentration leading to 50% cell-killing) of H-PEG decreased 5 fold (P<0.01) and 9 fold (P<0.01) against A2780 and HCT-8 cells compared with H-Lip, respectively. The results of biodistribution studies in sarcoma S(180) tumour bearing mice showed that liposomal encapsulation increased the concentration of total topotecan and the ratio of lactone form in plasma. H-PEG resulted in a 70 fold and 3.7-fold increase in AUC(0-->24 h) compared with free topotecan and H Lip, respectively. Moreover, H-PEG increased the accumulation of topotecan in tumours and the relative tumour uptake ratio compared with free topotecan was 5.2, and higher than that of H-Lip. The anti-cancer effect studies in murine heptocarcinoma H(22) tumour-bearing mice showed that H-PEG improved the therapeutic efficiency of topotecan and decreased the toxicity of topotecan to a certain extent compared with H-Lip. These results indicated that PEG-modified liposome might be an efficient carrier of topotecan. PMID- 16259757 TI - Nebulization of four commercially available amphotericin B formulations in persistently granulocytopenic rats with invasive pulmonary aspergillosis: evidence for long-term biological activity. AB - The nebulization of amphotericin B desoxycholate (AMB-DOC), liposomal amphotericin B (L-AMB), amphotericin B lipid complex (ABLC) and amphotericin B colloidal dispersion (ABCD) has been investigated. Particle sizes of generated aerosol droplets were measured. Pulmonary amphotericin B deposition and amphotericin B concentration in blood directly after nebulization and at six weeks after nebulization was measured in healthy rats. The efficacy of nebulized amphotericin B formulations was evaluated in persistently granulocytopenic rats with invasive pulmonary aspergillosis. Treatment was given either after or before fungal inoculation. The endpoint was survival of animals. Aerosol particle sizes, expressed as the values for the mass median diameter were 1.38, 2.43, 0.90 and 2.29 microm for AMB-DOC, L-AMB, ABLC and ABCD, respectively. Amphotericin B concentrations in the lungs directly after nebulization exceeded the minimum inhibitory concentration of Aspergillus fumigatus and amphotericin B was still detected in lungs of rats at six weeks after nebulization. Treatment, started at 16 h after fungal inoculation, resulted in a significantly prolonged survival as compared with sham-treated rats for all four formulations. Prophylactic treatment at one week before fungal inoculation resulted in a significantly prolonged survival for all four formulations. Aerosol treatment given at two weeks before inoculation was effective only for AMB-DOC and L-AMB, whereas treatment given at six weeks resulted in a significantly prolonged survival for L-AMB only. All commercially available amphotericin B preparations could be nebulized efficiently and may be of value in the prophylactic treatment of invasive pulmonary aspergillosis. PMID- 16259758 TI - Transport characteristics of rutin deca (H-) sulfonate sodium across Caco-2 cell monolayers. AB - The aim of this study was to explore potential transport mechanisms of rutin deca (H-) sulfonate sodium (RDS) across Caco-2 cell monolayers. As an in-vitro model of human intestinal epithelial membrane, Caco-2 cells were utilized to evaluate the transepithelial transport characteristics of this hydrophilic macromolecular compound. Bi-directional transport study of RDS demonstrated that the apparent permeability (P(app)) in the secretory direction was 1.4 approximately 4.5-fold greater than the corresponding absorptive P(app) at concentrations in the range 50.0 approximately 2,000 microM. The transport of RDS was shown to be concentration, temperature and pH dependent. In the presence of ciclosporin and verapamil, potent inhibitors of P-glycoprotein (P-gp)/MRP2, the absorptive transport was enhanced and secretory efflux was diminished. RDS significantly reduced the efflux ratio of the P-gp substrate rhodamine-123 in a fashion indicative of P-gp activity suppression, while rhodamine-123 competitively inhibited the polarized transport of the compound. In conclusion, the results indicated that RDS was likely a substrate of P-gp. Several efflux transporters, including P-gp, participated in the absorption and efflux of RDS and they might play significant roles in limiting the oral absorption of the compound. These observations offered important information for the pharmacokinetics of RDS. PMID- 16259759 TI - Transporter-mediated influx and efflux mechanisms of pitavastatin, a new inhibitor of HMG-CoA reductase. AB - The purpose of this study was to gain a better understanding of the transport mechanism of pitavastatin, a novel synthetic HMG-CoA reductase inhibitor. Experiments were performed using oocytes of Xenopus laevis expressing several solute carrier (SLC) transporters and recombinant membrane vesicles expressing several human ABC transporters. The acid form of pitavastatin was shown to be a substrate for human OATP1, OATP2, OATP8, OAT3 and NTCP, and for rat Oatp1 and Oatp4 with relatively low K(m) values. In contrast, these SLC transporters were not involved in the uptake of the lactone form. A significant stimulatory effect was exhibited by pitavastatin lactone, while the acid form did not exhibit ATPase hydrolysis of P-glycoprotein. In the case of breast cancer resistant protein (BCRP), the acid form of pitavastatin is a substrate, whereas the lactone form is not. Taking these results into consideration, several SLC and ABC transporters were identified as critical to the distribution and excretion of pitavastatin in the body. This study showed, for the first time, that acid and lactone forms of pitavastatin differ in substrate activity towards uptake and efflux transporters. These results will potentially contribute to the differences in the pharmacokinetic profiles of pitavastatin. PMID- 16259760 TI - Protective effect of taurine on myocardial antioxidant status in isoprenaline induced myocardial infarction in rats. AB - We have examined the protective effect of taurine on the myocardial antioxidant defense system in isoprenaline (isoproterenol)-induced myocardial infarction in rats, an animal model of myocardial infarction in man. Levels of diagnostic marker enzymes in plasma, lipid peroxides and reduced glutathione, and the activity of glutathione-dependent antioxidant enzymes and anti-peroxidative enzymes in the heart tissue were determined. Intraperitoneal administration of taurine significantly prevented the isoprenaline-induced increases in the levels of alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase, and creatine phosphokinase in the plasma of rats. Taurine exerted an antioxidant effect against isoprenaline-induced myocardial infarction by preventing the accumulation of lipid peroxides and by maintaining the level of reduced glutathione and the activity of glutathione peroxidase, glutathione-S transferase, catalase and superoxide dismutase at near normality. The results indicated that the cardioprotective potential of taurine was probably due to the increase of the activity of the free radical enzymes, or to a counteraction of free radicals by its antioxidant nature, or to a strengthening of myocardial membrane by its membrane stabilizing property. PMID- 16259761 TI - Design and synthesis of long-chain arylpiperazines with mixed affinity for serotonin transporter (SERT) and 5-HT(1A) receptor. AB - A new generation of antidepressant agents could be represented by compounds with mixed activity as serotonin transporter (SERT) inhibitors and 5-HT(1A) receptor antagonists. We report here on the synthesis and evaluation of SERT and 5-HT(1A) receptor affinity of long-chain arylpiperazines obtained either by modifying 6 nitroquipazine into a long-chain arylpiperazine or by inserting a modified 6 nitroquipazine moiety or other structures endowed with SERT affinity into a long chain arylpiperazine with 5-HT(1A) affinity. Among the compounds studied, 2-[4-(2 methoxyphenyl)piperazin-1-yl]-N-(6-nitro-2-quinolyl)ethylamine (21) and 1-(5 bromo-1,2,3,4-tetrahydronaphthalen-1-yl)-3-[4-(2-methoxyphenyl)-piperazin-1-yl]-1 propanone (24) showed good affinity values for SERT and 5-HT(1A) receptors (SERT: K(i) (inhibition constant)=71.8 and 62.8 nM; 5-HT(1A)K(i)=14.2 and 0.82 nM, respectively). PMID- 16259762 TI - Renal effects induced by the lectin from Vatairea macrocarpa seeds. AB - Lectins are glycoproteins that interact reversibly and specifically with carbohydrates. The renal effects of the galactose-binding lectin from the seeds of Vatairea macrocarpa were investigated. Isolated kidneys from Wistar rats (240 280 g) were perfused with Krebs-Henseleit solution containing 6% bovine serum albumin. The V. macrocarpa lectin (10 microg mL(-1)) increased the perfusion pressure, renal vascular resistance, urinary flow and glomerular filtration rate. However, V. macrocarpa lectin did not change the percentage sodium, potassium or chloride tubular transport. Pre-treatment with lectin-galactose complex significantly blocked the increase in perfusion pressure, renal vascular resistance, urinary flow and glomerular filtration rate. The control group showed a small amount of a proteinaceous material in the urinary space, although no alteration in the renal tubules was detected. The administration of galactose alone did not modify the functional parameters of the kidney. Kidneys perfused with V. macrocarpa lectin showed moderate deposits of a proteinaceous material in the tubules and urinary space. Those pre-treated with lectin-galactose complex had only small amount of a proteinaceous material in the urinary space. No abnormalities were seen in renal tubules. The results suggest that lectin from V. macrocarpa seeds has important effects on the carbohydrate-binding sites of the renal system, given the reversal of renal effects with the use of that specific inhibitor. PMID- 16259763 TI - The mechanisms underlying the anti-aging activity of the Chinese prescription Kangen-karyu in hydrogen peroxide-induced human fibroblasts. AB - Our previous study showed that Kangen-karyu extract protected against cellular senescence by reducing oxidative damage through the inhibition of reactive oxygen species generation and regulation of the antioxidative status. Although these findings suggest that Kangen-karyu could delay the aging process, the mechanisms responsible for protection against aging have rarely been elucidated. Therefore, this study was focussed on the mechanisms responsible for the anti-aging activity of Kangen-karyu extract using hydrogen peroxide (H(2)O(2))-induced human diploid fibroblasts, a well-established experimental model of cellular aging. Kangen karyu extract exerted a protective effect against the morphological changes induced by H(2)O(2) treatment and inhibited senescence-associated beta galactosidase activity. In addition, the beneficial effects of Kangen-karyu extract on cell viability and lifespan indicated that Kangen-karyu extract could delay the cellular aging process. The observation that Kangen-karyu extract prevented nuclear factor kappa B (NF-kappaB) translocation in response to oxidative stress suggested that Kangen-karyu exerted its anti-aging effect through NF-kappaB modulation and prevention of H(2)O(2)-induced overexpression of haem oxygenase-1 protein. Moreover, pretreatment with Kangen-karyu extract reduced overexpression of bax protein and prevented the mitochondrial membrane potential decline, suggesting that Kangen-karyu extract may protect mitochondria from mitochondrial oxidative stress and dysfunction. These findings indicate that Kangen-karyu is a promising potential anti-aging agent that may delay, or normalize, the aging process by virtue of its protective activity against oxidative stress-related conditions. PMID- 16259764 TI - Antinociceptive action of the extract and the flavonoid quercitrin isolated from Bauhinia microstachya leaves. AB - This study examined the antinociceptive effect of Bauhinia microstachya (Leguminosae), a native plant widely distributed in the South of Brazil, in several chemical and mechanical models of pain. The methanolic extract (ME) from B. microstachya (3--30 mg kg(-1), i.p.) and the isolated compound quercitrin (1- 10 mg kg(-1), i.p.), given 30 min earlier, produced a dose-dependent inhibition of acetic-acid-induced visceral pain in mice, with a mean ID50 value (dose necessary to reduce the nociceptive response by 50% relative to the control value) of 7.9 and 2.4 mg kg(-1), respectively. The ME of B. microstachya (3--100 mg kg(-1), i.p., 30 min earlier) also caused a dose-dependent inhibition of capsaicin-induced pain, with a mean ID50 value of 18.8 mg kg(-1). Moreover, the ME (3--100 mg kg(-1), i.p., 30 min earlier) produced marked inhibition of both phases of formalin-induced pain, with mean ID50 values for the neurogenic and the inflammatory phases of 30.3 and 17.2 mg kg(-1), respectively. In addition, the ME of B. microstachya (3--300 mg kg(-1), i.p., 30 min earlier) inhibited, in a graded manner, the hyperalgesia induced by bradykinin (3.2 microg/paw), substance P (13.5 microg/paw), carrageenan (300 microg/paw), capsaicin (100 microg/paw) and adrenaline (100 ng/paw) in the rat paw, with mean ID50 values of 20.5, 17.9, 101.8, 54.2 and 99.7 mg kg(-1), respectively. Taken together, these data demonstrate that ME of B. microstachya elicited a pronounced antinociceptive action against several chemical and mechanical models of pain in mice and rats. The precise mechanism responsible for the antinociceptive effect of the extract still remains unclear, but seems to be partly related to modulation of the release or action of pro-inflammatory mediators involved in the models of pain used. Finally, the flavonoid quercitrin isolated from this plant appears to contribute for the antinociceptive property of the methanolic extract. PMID- 16259765 TI - Preventive effect of Aegle marmelos leaf extract on isoprenaline-induced myocardial infarction in rats: biochemical evidence. AB - We have evaluated the preventive effects of an aqueous Aegle marmelos leaf extract (AMLEt) in isoprenaline (isoproterenol)-induced myocardial infarction in rats. Rats were pretreated with AMLEt (50, 100 or 200 mg kg(-1)) for 35 days. After the treatment period, isoprenaline (200 mg kg(-1)) was administered subcutaneously to rats at an interval of 24 h for two days. The activity of creatine kinase (CK) and lactate dehydrogenase (LDH) was significantly increased in serum and significantly decreased in heart of isoprenaline-treated rats. Pretreatment with AMLEt decreased the activity of CK and LDH in serum and increased them in the heart. The activity of sodium-potassium dependent adenosine triphosphatase (Na(+)K(+)ATPase) was significantly decreased while the activity of calcium dependent adenosine triphosphatase (Ca(2+)ATPase) was simultaneously increased in the heart and aorta. AMLEt pretreatment increased the activity of Na(+)K(+) ATPase and decreased the activity of Ca(2+)ATPase in the heart and aorta simultaneously. The levels of cholesterol and triglycerides increased, while the levels of phospholipids decreased in the heart and aorta of isoprenaline-treated rats. In AMLEt-pretreated rats the levels of cholesterol and triglycerides decreased whereas phospholipids increased in heart and aorta. All the deranged biochemical parameters were restored with 200 mg kg(-1) AMLEt. Similarly alpha-tocopherol (60 mg kg(-1))-pretreatment to isoprenaline-treated rats exhibited a significant effect on all the parameters studied. The results from this study may have clinical relevance. PMID- 16259766 TI - Bioavailability of glycyrrhizin from Shaoyao-Gancao-Tang in laxative-treated rats. AB - Shaoyao-Gancao-Tang (SGT), a traditional Chinese formulation composed of Shaoyao (Paeoniae Radix) and Gancao (Glycyrrhizae Radix), is frequently used in conjunction with laxatives such as sodium picosulfate in colonoscopy to relieve abdominal pains. We have investigated the alterations of the bioavailability of glycyrrhizin when SGT was co-administered with sodium picosulfate and we tried to identify a regimen that might minimize the alterations. Glycyrrhizin is one of the active glycosides in Gancao and SGT and is hydrolysed into the bioactive metabolite, 18 beta-glycyrrhetic acid (GA) by intestinal bacteria following oral administration. We found that the maximum plasma concentration (C(max)) and the area under the mean concentration vs time curve from zero to 24 h (AUC(0-24 h)) of GA from a single dose of SGT administered 5 h after a single pretreatment with sodium picosulfate were significantly reduced to 15% and 20% of the control level in rats, respectively. These reductions were still significant four days after sodium picosulfate pretreatment, but were restored by repetitive administration of SGT following sodium picosulfate pretreatment. Similar reductions and recovery were observed for the glycyrrhizin-metabolizing activity of intestinal bacteria in rat faeces. The results warrant clinical studies for co-administration of laxatives such as sodium picosulfate and SGT. PMID- 16259767 TI - Oral administration of a decaffeinated green tea (Camellia sinensis) extract did not alter urinary 8-epi-prostaglandin F(2 alpha), a biomarker for in-vivo lipid peroxidation. AB - Oxidative stress is involved in the pathogenesis of numerous chronic human diseases. The objective of this study was to determine whether administration of a decaffeinated green tea extract providing 844 mg flavonoids daily reduced the urinary excretion of 8-epi-prostaglandin F(2 alpha) (8-epi-PGF(2 alpha)), a product of lipid peroxidation in cellular membranes and of low-density lipoprotein (LDL). Nine healthy male and female subjects were studied at baseline and after 14 days of green tea supplementation. Analysis of urinary 8-epi-PGF(2 alpha) was performed using immunoaffinity extraction-gas chromatography-negative ion chemical ionization-mass spectrometry (GC-NICI-MS). Urinary 8-epi-PGF(2 alpha) concentrations were 0.286+/-0.120 nmol (mmol creatinine)(-1) at baseline and 0.244+/-0.177 nmol mmol(-1) creatinine after green tea supplementation. There were no significant differences in the excretion of urinary 8-epi-PGF(2 alpha) after treatment with green tea. We conclude that 14 days of green tea supplementation did not significantly alter in-vivo lipid peroxidation. PMID- 16259768 TI - Respiratory syncytial virus infections: characteristics and treatment. AB - In this review, we describe the history, epidemiology and clinical manifestations of infections attributed to respiratory syncytial virus (RSV) in children. At present, no cure exists for RSV infection but commonly employed palliative treatments include oxygen and inhaled beta(2)-adrenoceptor agonists, such as salbutamol, to relieve the wheezing and increased bronchiolar smooth muscle constriction. Adrenaline (epinephrine) has been found to be superior to the selective beta(2)-adrenoceptor agonists. Oral or inhaled corticosteroids should counteract the inflammatory response to RSV infection but their effectiveness is controversial. Inhaled ribavirin is the only licensed antiviral product approved for the treatment of RSV lower respiratory-tract infection in hospitalized children, although its use is now restricted to high-risk infants. Other treatments considered are nasopharyngeal suctioning, surfactant therapy, recombinant human deoxyribonuclease I, heliox (helium:oxygen) and inhaled nitric oxide. Prevention of infection by RSV antibodies is another strategy and, currently, palivizumab is the only safe, effective and convenient preventative treatment for RSV disease in high-risk populations of infants and young children. Its cost-effectiveness, however, has been questioned. Both live attenuated and subunit vaccines against RSV infection have been developed but so far there is no safe and effective vaccine available. Finding effective treatments and prophylactic measures remains a major challenge for the future. PMID- 16259769 TI - Towards an understanding of adsorption behaviour in non-aqueous systems: adsorption of poly(vinyl pyrrolidone) and poly(ethylene glycol) onto silica from 2H, 3H-perfluoropentane. AB - The adsorption behaviour of low molecular weight poly(ethylene glycol) (PEG 600) and poly(vinyl pyrrolidone) (PVP K25) to silica particles has been investigated at room temperature (21 degrees C) in the partially fluorinated solvent 2H,3H perfluoropentane (HPFP). PVP (absorbed amount, Gamma = 12 mg g(-1)) was found to adsorb more strongly than PEG (Gamma = 4 mg g(-1)). Both of these values were higher than observed in water. In a further distinction to the aqueous case, where PVP displaces PEG from the interface, no competitive adsorption effects were observed between these two polymers in HPFP, with the adsorbed amounts of each polymer being unchanged by the presence of the other. The stability of silica suspensions in HPFP was primarily dependent on the presence of PVP; PEG/silica systems were unstable, but PVP/silica and PEG/PVP/silica systems formed stable suspensions. All suspensions were destabilized by the addition of small (0.15 wt%) amounts of water. The observations made in this work would point to a flocculation phenomenon due to the addition of water, and not Ostwald ripening. The mechanism of this destabilization is likely to be water acting as a flocculation bridge between particles. PMID- 16259770 TI - Discriminant analysis as a tool to identify compounds with potential as transdermal enhancers. AB - Structure-activity relationships were sought for 73 enhancers of hydrocortisone permeation from propylene glycol across hairless mouse skin. Enhancers had chain lengths (CC) from 0 to 16 carbon atoms, 1 to 8 H-bonding atoms (HB), molecular weight 60 to 450, log P (calculated) -1.7 to 9.7 and log S (calculated) -7.8 to 0.7. These predictive properties were chosen because of their ready availability. Enhancement ratio (ER) was defined as hydrocortisone transferred after 24 h relative to control. Values for the ER ranged from 0.2 to 25.3. Multiple regression analysis failed to predict activity; ER values for the 'good' enhancers (ER > 10) were underestimated. Simple guidelines suggested that high ER was associated with CC > 12 and HB 2-5. This was refined by multivariate analysis to identify significant predictors. Discriminant analysis using CC, HB, and molecular weight correctly assigned 11 of the 12 'good' enhancers (92%). The incorrectly assigned compound was a known, idiosyncratic Br compound. Seventeen of the 61 'poor' enhancers (28%) were incorrectly assigned but four could be considered marginal (ER > 8). The success of this simple approach in identifying potent enhancers suggested its potential in predicting novel enhancer activity. PMID- 16259771 TI - Pharmacokinetic interaction between 5-[2-propyloxy-5-(1-methyl-2 pyrollidinylethylamidosulfonyl)phenyl]-1-methyl-3-propyl-1,6-dihydro-7H-pyrazolo (4,3-d) pyrimidine-7-one (DA-8159) and nitroglycerin in rats. AB - The pharmacokinetic interaction between 5-[2-propyloxy-5-(1-methyl-2 pyrollidinylethylamidosulfonyl)phenyl]-1-methyl-3-propyl-1,6-dihydro-7H-pyrazolo (4,3-d)pyrimidine-7-one (DA-8159), a new erectogenic, and nitroglycerin has been evaluated in rats. Male Sprague-Dawley rats received DA-8159 (30 mg kg(-1)) as a single intravenous or oral dose with the simultaneous single intravenous administration of nitroglycerin (2.5 mg kg(-1)). After simultaneous intravenous administration, the total area under the plasma concentration-time curve from time zero to time infinity (AUC(inf)) of DA-8159 (746 vs 457 microg min mL(-1)) was found to be significantly greater than with DA-8159 alone. Also, after simultaneous intravenous administration total body clearance (CL) (40.2 vs 65.6 mL min(-1) kg(-1)), renal clearance (CL(R)) (1.65 vs 5.11 mL min(-1) kg(-1)), and nonrenal clearance (CL(NR)) (38.3 vs 60.2 mL min(-1) kg(-1)) of DA-8159 were significantly slower compared with DA-8159 alone. The slower CL(NR) of DA-8159 could have been due to the inhibition of the metabolism of DA-8159 by nitroglycerin, since DA-8159 is metabolized via CYP3A1/2 in rats and nitroglycerin inhibits CYP3A1/2 in rats. The slower CL(R) of DA-8159 could have been due to the urine flow rate-dependent CL(R) of DA-8159 in rats. After the simultaneous intravenous administration of nitroglycerin and DA-8159, the AUC(inf) of nitroglycerin was significantly smaller (635 vs 960 microg min mL( 1)), which could have been due to the cardiac output-dependent CL of nitroglycerin. However, after the oral administration of DA-8159, the pharmacokinetic parameters of DA-8159 with and without the intravenous administration of nitroglycerin became comparable. This was not due to the decrease in nitroglycerin's gastrointestinal absorption of DA-8159, but could have been due to changes in nitroglycerin's intestinal firstpass effect of DA 8159. Human studies are required to determine the administration time of DA-8159 when nitroglycerin is concomitantly taken. PMID- 16259772 TI - The pharmacokinetics of etanercept in patients with end-stage renal disease on haemodialysis. AB - Inflammation is strongly associated with malnutrition and cardiovascular risk in patients with chronic renal failure on haemodialysis (HD). The acute-phase inflammatory response, defined by the increased synthesis of positive acute-phase proteins, is stimulated by the production of such cytokines as interleukin 6 (IL 6), interleukin 1 (IL-1) and tumour necrosis factor-alpha TNF-alpha The availability of cytokine antagonists allows testing of the hypothesis that suppression of inflammation reverses the malnutrition-inflammation syndrome in HD patients. Etanercept is a soluble TNF-alpha receptor fusion protein used to suppress inflammation in rheumatoid and psoriatic arthritis. Its metabolism in HD patients is unknown. In a study designed to test the safety and pharmacokinetics of etanercept in HD patients, etanercept was administered to six HD patients with albumin levels above 4.2 g dL(-1) and C-reactive protein levels <5 mg L(-1) (five men, one woman, age range 34-59 years). Etanercept (25 mg) was administered subcutaneously twice weekly immediately after dialysis for 13-16 weeks. Etanercept concentrations were measured pre- and post-dialysis by ELISA. Concentrations were compared graphically to assess whether, firstly, dialysis affects etanercept apparent clearance and, secondly, etanercept kinetics were similar between HD patients and the more extensively studied psoriasis population with normal renal function (PS). The second stage examined model-based parameter predictions of the terminal elimination rate constant (k) for HD patients. Steady state etanercept levels were comparable between HD and PS patients. Treatment with HD had no effect on etanercept levels. When etanercept was discontinued, the terminal rate constant for HD patients was not significantly different from that observed in PS patients. No adverse effects were noted during the 3-month treatment phase and subsequent 6-month follow-up. Albumin and C-reactive protein levels did not change in these non-inflamed patients during the study period. The pharmacokinetics of etanercept in patients with chronic renal failure on HD are similar to patients with normal renal function. It is, therefore, feasible to administer etanercept to HD patients without adjusting the dose. PMID- 16259773 TI - Dexmedetomidine prolongs spinal anaesthesia induced by levobupivacaine 0.5% in guinea-pigs. AB - Alpha-2 adrenoceptor agonists have been used in association with local anaesthetic to increase the duration of spinal anaesthesia. Intrathecal administration of clonidine prolonged motor blockade induced by local anaesthetic. Since the affinity of dexmedetomidine (DEX) to alpha-2 adrenoceptors is eight-times greater than clonidine, it is expected that DEX could be advantageous in clinical anaesthesia. We investigated the duration of motor nerve block induced by spinal injection of 0.5% levobupivacaine (LVB) associated with intrathecal or intraperitoneal administration of DEX. Seventy-two guinea-pigs were randomly divided in 12 groups, which were all treated with intrathecal injection of 50 microL of LVB. DEX was injected intrathecally with LVB in 6 groups or injected intraperitoneally after LVB in another 6 groups. Intrathecal DEX (0.1, 0.2 and 0.4 microg) increased the LVB-induced motor anaesthesia from 48 (41-66) min to 84.5 (52-91) min (P < 0.05), 101.5 (83-115) min (P < 0.05) and 105 (97-114) min (P < 0.05), respectively. Similarly, intraperitoneal DEX (20 and 40 microg kg(-1)) increased the motor blockade from 48.5 (33-59) min to 88 (71-114) min (P < 0.05) and 114.5 (103-156) min (P < 0.05), respectively. Pre-treatment with yohimbine reduced the duration of motor block from 101.5 (83-115) to 76.5 (68-86) min (P < 0.05) or from 114.5 (103-156) to 90 (83-93) min (P < 0.05) when DEX was administered by the intrathecal or intraperitoneal routes. Motor block induced by spinal injection of LVB was prolonged by intrathecal and systemic administration of DEX, which was partially dependent on activation of alpha-2 adrenoceptors. PMID- 16259774 TI - Involvement of P-glycoprotein in the release of cytokines from peripheral blood mononuclear cells treated with methotrexate and dexamethasone. AB - P-glycoprotein (P-gp), a product of the MDR1 gene, is an important factor in the turnover of many drugs and xenobiotics. Recent reports have suggested that P-gp can also be involved in the transport of cytokines. The aim of this study was to examine the role of P-gp in cytokine release from phytohaemagglutinin (PHA) stimulated peripheral blood mononuclear cells (MNCs) as well as in the release of cytokines from MNCs treated with methotrexate (MTX) and dexamethasone (DEX). The study was carried out on PHA-stimulated MNC from 10 healthy subjects. Flow cytometry was applied to measure interleukin (IL)-2, IL-4, IL-6, IL-10, interferon (IFN)-gamma and tumour necrosis factor (TNF)-alpha levels in the culture supernatants. In the experiments verapamil (VER) and P-gp specific monoclonal antibodies (mAb) (clone 17F9) were used to inhibit P-gp function. P-gp inhibitors suppressed the release of IL-2, IL-4, IFN-gamma and TNF-alpha from PHA stimulated MNC, whereas release of IL-6 and IL-10 remained unaffected. VER and mAb significantly decreased the release of IL-2, IL-4, TNF-alpha and INF-gamma in MNC cultures treated with MTX or DEX. The results of this study suggest that P-gp may be involved in the transmembrane transport of some cytokines. Moreover, it seems that blocking of P-gp function may influence the release of some cytokines from MNCs, displaying an additive inhibitory effect to DEX and MTX. PMID- 16259775 TI - Synthesis and pharmacological effects of new, N-substituted soft anticholinergics based on glycopyrrolate. AB - To reduce the possibility of systemic side-effects in locally administered anticholinergics, two new N-substituted glycopyrrolate analogues designed using soft drug design approaches have been synthesized and evaluated in vitro and in vivo. Because stereospecificity is known to be important at muscarinic receptors, the new compounds SGM and SGE also have been prepared as their pure 2R isomers, 2R-SGM and 2R-SGE, by starting from optically pure (-)-cyclopentylmandelic acid, and the corresponding isomers were indeed found to be more active. The new soft glycopyrrolates were chemically more stable under acidic conditions, and the ethyl esters SGE were more stable than the methyl esters SGM. The new compounds were also found to be quite susceptible to extrahepatic metabolism, having half lives of 20-30 min in rat plasma (in vitro), consistent with their soft nature. Binding studies at human muscarinic receptors (M(1)-M(4)) and guinea-pig ileum assays found 2R-SGM and 2R-SGE to have potencies somewhat less than, but close to, those of glycopyrrolate and N-methylscopolamine. They caused pupil dilation in rabbit eyes, but their mydriatic effects lasted for considerably less time than that of glycopyrrolate, and they did not induce dilation of the pupil in the contralateral, water-treated eyes, indicating that, in agreement with their soft nature, they are locally active, but safe and with a low potential to cause systemic side-effects. PMID- 16259776 TI - Role of lupeol and its ester on cyclophosphamide-induced hyperlipidaemic cardiomyopathy in rats. AB - Cyclophosphamide, an alkylating agent widely used in cancer chemotherapy, causes fatal cardiotoxicity. In this study, lupeol, a pentacyclic triterpene isolated from Crataeva nurvala stem bark, and its ester, lupeol linoleate, were investigated for their possible hypocholesterolaemic effects against cyclophosphamide-induced lipidaemic instabilities. Male albino Wistar rats were categorized into 6 groups. Group I served as control. Rats in groups II, V and VI were injected intraperitoneally with a single dose of cyclophosphamide (200 mg kg(-1)) dissolved in saline. Cyclophosphamide-treated groups V and VI respectively received lupeol and lupeol linoleate (50 mg kg(-1)), dissolved in olive oil, for 10 days by oral gavage. Groups III and IV served as drug controls and were administered lupeol and lupeol linoleate, respectively. Cyclophosphamide administration induced abnormal changes in serum lipoproteins and lipid fractions in both serum and cardiac tissue. The activity of lipid metabolizing enzymes was distorted significantly in the cyclophosphamide-treated rats. The cyclophosphamide-treated rats also showed extensive intermuscular haemorrhage in histology. Lupeol and its ester reversed the above alterations induced by cyclophosphamide. This study encapsulates the early lipaemic abnormalities in the heart tissue of cyclophosphamide-treated rats. Treatment with lupeol linoleate was more effective than lupeol in rendering protection to the cardiac tissue challenged by cyclophosphamide. PMID- 16259777 TI - Dietary palm olein oil augments cardiac antioxidant enzymes and protects against isoproterenol-induced myocardial necrosis in rats. AB - Wistar rats, 150-200 g, of either sex, were fed daily with commercial rat diet supplemented with palm olein oil in two doses (5% v/w (n = 16) and 10% v/w (n = 16) of diet) for 30 days. Control rats (n = 16) were fed with normal diet. On the 29(th) and 30(th) days, 8 rats from each group were administred isoproterenol (85 mg/kg, s.c., 24-h interval). On the 31(st) day, all rats were sacrificed and myocardial tissues were studied for thiobarbituric acid reactive substances (TBARS), antioxidant enzymes and light microscopic changes, along with the ferric reducing ability of plasma (FRAP). A significant rise in myocardial superoxide dismutase (SOD), catalase and glutathione peroxidase (GPx) activity and FRAP level were observed in rats fed with palm olein oil. Isoproterenol caused an increase in myocardial oxidative stress in control rats, as evidenced by an increase in myocardial TBARS level, reduction in FRAP and myocardial SOD, catalase and GPx activity, along with focal necrosis of cardiac muscle fibres on light microscopy. The rise in myocardial TBARS and depletion of SOD and catalase activity following isoproterenol administration were prevented in palm-olein-oil supplemented diet-fed rats at both doses. Isoproterenol-induced myocardial light microscopic changes were also prevented in the treated groups. The results suggest that dietary palm olein oil caused augmentation of myocardial antioxidant enzymes and protected against isoproterenol-induced myocardial necrosis and associated oxidative stress. PMID- 16259779 TI - Treatment with Y-40138, a multiple cytokine production modulator, inhibits lipopolysaccharide- or tumour necrosis factor-alpha-induced production of pro inflammatory cytokines and augments interleukin-10. AB - N-[1-(4-[4-(pyrimidin-2-yl)piperazin-1-yl]methyl phenyl)cyclopropyl] acetamide . HCl (Y-40138) suppresses liver injury in concanavalin A- and D galactosamine/lipopolysaccharide (LPS)-induced mouse hepatitis models. However, the mechanism of action of Y-40138 has not been fully investigated. In this study, we examined the effect of Y-40138 on cytokine production in mice. Cytokine production was induced by intraperitoneal injection of LPS (0.5 mg kg(-1)) or intravenous injection of recombinant mouse tumour necrosis factor (TNF)-alpha (10 mug mouse(-1)) in BALB/c mice. TNF-alpha and interleukin (IL)-10 reached maximum levels 1.5 h after the LPS injection. IL-12 and interferon-sigma (IFN-sigma) reached maximum levels 3 to 9 h after the injection. When Y-40138 was orally administered 30 min prior to the injection, it inhibited TNF-alpha, IL-12 and IFN sigma production and augmented IL-10 production. Y-40138 also inhibited IL-12 production and augmented IL-10 production in TNF-alpha-stimulated mice. In IL-10 knockout mice, Y-40138 inhibited TNF-alpha and IL-12 production 1.5 h after the LPS injection but not after 3 h or later, unlike in wild mice. In addition, TNF alpha production was inhibited by Y-40138 at concentrations that could not augment IL-10 production. These data suggest that Y-40138 modulates pro inflammatory cytokine production by both IL-10-dependent and -independent mechanisms. PMID- 16259778 TI - Distribution of sigma receptors in EMT-6 cells: preliminary biological evaluation of PB167 and potential for in-vivo PET. AB - Sigma(1) and sigma(2) receptors have been detected in many tissues and are highly expressed in several tumour cell lines from various tissues. The high level of expression observed for sigma receptors and their involvement in cell proliferation and apoptosis has led to the development of several sigma ligands in order to obtain a molecular probe for in-vivo diagnostic imaging techniques such as positron emission tomography (PET) and single photon emission computerized tomography (SPECT). The EMT-6 cells implanted in mice were a good model for evaluating the proliferation of solid tumours by in-vivo PET. Moreover, we developed the sigma ligand PB167, a cyclohexylpiperazine derivative, previously evaluated for sigma(2) receptor affinity and activity in standard protocols. The related results encouraged us to verify if this compound could be developed as a radiotracer for in-vivo PET in order to visualize sigma(2) receptors expressed in EMT-6 cells when implanted in mice. This perspective was thought to be favourable because PB167 bears a methoxy substituent on the tetraline nucleus, an easy point for (11)C labelling. The aims of this preliminary study were both to assess the relative distribution of sigma(1) and sigma(2) receptors in EMT-6 cells and to verify if PB167 could be developed as a sigma(2) radiotracer for in-vivo PET. The results showed that both sigma(1) and sigma(2) receptors were overexpressed in EMT-6 cells and that the ligand PB167 can be positively considered for radiosynthesis preparation in order to suitably visualize sigma(2) receptors by the in-vivo PET technique and correlate their presence to tumour proliferation. PMID- 16259780 TI - Dietary fatty acid unsaturation levels, lipoprotein oxidation and circulating chemokine in experimentally induced atherosclerotic rats. AB - The dietary balance of long-chain fatty acids may influence processes involving leukocyte endothelial interactions, such as atherogenesis and inflammation. The relationship between proatherogenic lipoproteins and chemotactic motility is still controversial. However, the interaction of the former can increase recruitment of monocytes to the vessel walls and accelerate the events of atherogenesis. The current study examined the effects of unsaturated fatty acid levels on the oxidative susceptibility of lipoprotein, chemokine expressions and their relationship to atherosclerotic lesion development in experimental rats. Male Wistar rats were fed an atherogenic diet for 4 months and the diet was then supplemented with 10% v/w of virgin olive oil (OO group), sunflower oil (SO group) or fish oil (FO group) for 4 and 8 weeks. Blood samples were collected at four time points: at baseline, after feeding with the atherogenic diet and during the dietary regimen (4 and 8 weeks). Plasma lipid profile and lipoprotein oxidative susceptibility (LOS), C-reactive protein (CRP), monocyte chemoattractant protein (MCP-1), and regulated upon activation normal T-cell expressed and secreted (RANTES) were measured. The superoxide dismutase (SOD) and reduced glutathione (GSH) antioxidant activities were also studied in aortic segments. Histological assessment of the aortic segment was determined. Compared to baseline data, the high-fat and cholesterol-enriched diet increased atheroma formation, plasma LOS and inflammatory indexes (CRP, MCP-1, RANTES). However, it dramatically reduced aortic SOD and GSH contents. Dietary treatment of atherosclerotic rats with OO greatly reduced LOS and remarkably increased aortic SOD and GSH contents as compared to the SO- and FO-treated groups. The FO supplemented diet had a more pronounced lowering effect on MCP-1 and RANTES compared to the OO and SO diets. In conclusion, this study demonstrated a strong relationship between LOS and circulating levels of chemokines. OO is a potent antioxidant and moderate anti-inflammatory, which effectively reduced aortic atherosclerotic lesions more than the SO- or FO-treated groups in male Wistar rats. PMID- 16259781 TI - Statin-induced apoptosis linked with membrane farnesylated Ras small G protein depletion, rather than geranylated Rho protein. AB - Rhabdomyolysis is a severe adverse effect of 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors (statins). This myopathy is strongly enhanced by the combination with statins and fibrates, another hypolipidaemic agent. We have evaluated the initial step of statin-induced apoptosis by the detection of membrane flip-flop using flow cytometric analysis. L6 rat myoblasts were treated with various statins (atorvastatin (3 microM), cerivastatin (3 microM), fluvastatin (3 microM), pravastatin (3 mM), or simvastatin (3 microM)) for 2, 4 or 6 h followed by reacting with FITC-conjugated annexin V for the detection of initial apoptosis signal (flip-flop). Various statin-treated myoblasts were significantly stained with FITC-annexin V at 6 h, whereas they were not detected at 2 h. Moreover, immunoblot analysis indicated that when the cells were treated with cerivastatin (3 microM), membrane-associated Ras protein was activated and detached until 6 h, resulting in cell death through the consequent activation of caspase-8. On the other hand, since cytosolic Ras activation did not activate, there is still an unknown mechanism in statin-related Ras depletion. In conclusion, statin-induced apoptosis in muscular tissue was directly initiated by the farnesyl-anchored Ras protein depletion from cell membrane with subsequent apoptosis. PMID- 16259782 TI - Attenuation of ciclosporin-induced nephrotoxicity by dietary supplementation of seal oil in Sprague-Dawley rats. AB - Fish oil, rich in omega-3 (n-3) polyunsaturated fatty acids (PUFAs), has been reported to attenuate nephrotoxicity induced by ciclosporin (cyclosporine A). Harp seal oil is a rich source of n-3 PUFAs. This study investigated the ability of dietary seal oil to reduce nephrotoxicity caused by ciclosporin. Sprague Dawley rats were maintained on a standard diet (with sunflower oil as lipid, SFO) or a diet enriched with seal oil (with 85% seal oil and 15% sunflower oil as lipid, SO) for four weeks before and four weeks after intravenous administration of ciclosporin (15 mg kg(-1) daily). Kidney function was assessed by measuring blood urea nitrogen, creatinine clearance, urinary N-acetyl-1-beta-D glucosaminidase, 6-keto-prostaglandin F(1alpha), thromboxane B(2) and malondialdehyde. Systolic blood pressure (SBP) was monitored. Ciclosporin concentrations in blood were measured using liquid chromatographytandem mass spectrometry (LC-MS/MS). The fatty acid compositions of the diets and erythrocyte membranes were analysed by gas chromatography (GC). The results showed that nephrotoxicity was induced by ciclosporin in rats maintained on both SO and SFO diets. However, rats fed on SO diet endured less toxicity than those on SFO diet. The n-3 and n-6 PUFAs in the erythrocyte membrane of rats maintained on SO diet were found to be 10.79% and 11.93%, while those in rats maintained on SFO diet were found to be 1.67% and 22.71%, respectively. In conclusion, dietary supplementation of seal oil was found to reduce ciclosporin-induced nephrotoxicity in rats. PMID- 16259784 TI - Protective effects of Polygala paniculata extract against methylmercury-induced neurotoxicity in mice. AB - We have examined the possible protective effects of Polygala paniculata extract against methylmercury (MeHg)-induced neurotoxicity in adult mice. MeHg was diluted in drinking water (40 mg L(-1), freely available) and the hydroalcoholic Polygala extract was diluted in a 150 mM NaCl solution and administered by gavage (100 mg kg(-1) b.w., twice a day). After a two-week treatment, MeHg exposure significantly inhibited glutathione peroxidase and increased glutathione reductase activity, while the levels of thiobarbituric acid reactive substances were increased in the cerebral cortex and cerebellum. These alterations were prevented by administration of Polygala extract, except for glutathione reductase activity, which remained elevated in the cerebral cortex. Behavioural interference in the MeHg-exposed animals was evident through a marked deficit in the motor performance in the rotarod task, which was completely recovered to control levels by Polygala extract co-administration. This study has shown, for the first time, the in-vivo protective effects of Polygala extract against MeHg induced neurotoxicity. In addition, our findings encourage studies concerning the beneficial effects of P. paniculata on neurological conditions related to excitotoxicity and oxidative stress. PMID- 16259783 TI - Species differences in the gut stimulatory effects of radish seeds. AB - This study describes the gastrointestinal (GI) prokinetic effects of the aqueous extract of radish seeds (Rs.Cr). Rs.Cr, which tested positive for terpenes, flavonoids, phenols, alkaloids and saponins, showed a spasmogenic effect in isolated rabbit jejunum and ileum, rat stomach fundus and ileum, and guinea-pig ileum and jejunum. Rs.Cr was around 10 times more potent in the guinea-pig tissues and this effect was resistant to atropine, pyrilamine or SB203186 while the spasmogenic effect in the rat and rabbit tissues was atropine sensitive. The extract exhibited atropine-sensitive GI prokinetic and laxative effects in vivo in mice. In the atropinized rabbit jejunum, Rs.Cr produced a spasmolytic effect independent of Ca(++) or K(+) channels, adrenergic or opioid receptor involvement. Activity-directed fractionation of Rs.Cr yielded four fractions, all showing effects similar to that of the parent extract. Rs.Cr and its fractions were found to be non-lethal up to 10 g kg(-1) in mice for 24 h, except for the petroleum fraction, which showed 50% mortality at high doses. Some known radish compounds (spermine, spermidine, putrescine and sinigrin) were also tested and found to be devoid of any activity. The study shows species-specific spasmogenic effects of radish in rabbit, rat and mouse via muscarinic receptors but through an uncharacterized pathway in guinea-pig tissues. Additionally, a dormant relaxant effect was also seen, while the three polyamines and one glucosinolate from radish were found to be inactive, indicating that the compound(s) responsible for the activities reported remains to be isolated. PMID- 16259786 TI - Oxidative stress in myocardial ischaemia reperfusion injury: a renewed focus on a long-standing area of heart research. AB - Advances in the treatment of coronary artery disease have seen a significant drop in mortality and morbidity particularly amongst patients with acute myocardial infarction (MI). In particular, percutaneous trans-luminal balloon angioplasty (PTCA) with stenting to re-open atherosclerotic coronary arteries has yielded marked improvement in clinical outcome for patients with acute MI. Furthermore, with the advent of drug-eluting stents occurrence rates for coronary artery restenosis, one common clinical problem associated with angioplasty and stent deployment, have declined markedly. However, coronary restenosis in diabetic patients remains an on-going problem. The success of drug-eluting stents has seen a renewed focus on myocardial ischaemia reperfusion (IR) injury as this represents one area of research where many questions remain unanswered. In particular, the relationship between myocardial IR injury and decreased myocardial micro-vasculature re-flow post PTCA (that ultimately leads to poor clinical outcome and myocardial damage/dysfunction) is one area of research with the potential to decrease current complication rates further in patients suffering myocardial IR injury sustained during MI. This review discusses the role for oxidative stress, oxidant source(s) and both gene regulation and stem cell therapy as potential strategic targets in the ischaemic myocardium, with the ultimate aim of providing significant cardioprotection in the setting of acute MI. PMID- 16259785 TI - Oxidized proteins and their contribution to redox homeostasis. AB - Proteins are major target for radicals and other oxidants when these are formed in both intra- and extracellular environments in vivo. Formation of lesions on proteins may be highly sensitive protein-based biomarkers for oxidative damage in mammalian systems. Oxidized proteins are often functionally inactive and their unfolding is associated with enhanced susceptibility to proteinases. ROS scavenging activities of intact proteins are weaker than those of misfolded proteins or equivalent concentrations of their constituent amino acids. Protein oxidation and enhanced proteolytic degradation, therefore, have been suggested to cause a net increase in ROS scavenging capacity. However, certain oxidized proteins are poorly handled by cells, and together with possible alterations in the rate of production of oxidized proteins, may contribute to the observed accumulation and damaging actions of oxidized proteins during ageing and in pathologies such as diabetes, arteriosclerosis and neurodegenerative diseases. Protein oxidation may play a controlling role in cellular remodelling and cell growth. There is some evidence that antioxidant supplementation may protect against protein oxidation, but additional controlled studies of antioxidant intake to evaluate the significance of dietary/pharmacological antioxidants in preventing physiological/pathological oxidative changes are needed. PMID- 16259787 TI - Lysosomal ROS formation. AB - Ubiquinone is inhomogenously distributed in subcellular biomembranes. Apart from mitochondria, where ubiquinone has bioenergetic and pathophysiological functions, unusually high levels of ubiquinone have also been reported in Golgi vesicles and lysosomes. In lysosomes, the interior differs from other organelles in its low pH value which is important to ensure optimal activity of hydrolytic enzymes. Since redox-cycling of ubiquinone is associated with the acceptance and release of protons, we assumed that ubiquinone is part of a redox chain contributing to unilateral proton distribution. A similar function of ubiquinone was earlier suggested by Crane to operate in Golgi vesicles. Support for the involvement of ubiquinone in a presumed couple of redox carriers came from our observation that almost 70% of total lysosomal ubiquinone was in the divalently reduced state. Further reduction was seen in the presence of external NADH. Analysis of the components involved in the transfer of reducing equivalents from cytosolic NADH to ubiquinone revealed the existence of an FAD-containing NADH dehydrogenase. The latter was found to reduce ubiquinone by means of a b-type cytochrome. Proton translocation into the interior was linked to the activity of the novel lysosomal redox chain. Oxygen was found to be the terminal electron acceptor, thereby also regulating acidification of the lysosomal matrix. In contrast to mitochondrial respiration, oxygen was only trivalently reduced giving rise to the release of HO radicals. The role of this novel proton-pumping redox chain and the significance of the associated ROS formation has to be elucidated. PMID- 16259788 TI - Chronically active: activation of microglial proteolysis in ageing and neurodegeneration. AB - One of the microglial cell functions is the removal of modified extracellular proteins in the brain. The connection between protein oxidation, proteolysis, and microglial activation is the topic of this review. The effect of various activation agents on microglial cells with regard to changes in substrate uptake, proteolytic capacity and degradation efficiency of different types of oxidized protein materials is reviewed. It is shown that different activation stimuli initiate substrate-specific modulation for uptake and proteolysis, influencing an array of factors including receptor expression, lysosomal pH, and proteasome subunit composition. Age-related alterations in activation and proteolytic capacity in microglial cells are also discussed. In ageing, proteolytic effectiveness is diminished, while microglial cells are chronically activated and lose the oxidative burst ability, possibly supporting a 'vicious circle' of macrophage-induced neurodegeneration. PMID- 16259789 TI - Effects of antioxidants on glutathione levels and clinical recovery from the malnutrition syndrome kwashiorkor--a pilot study. AB - Kwashiorkor is a severe edematous form of malnutrition with high prevalence and lethality in many African countries, and repeatedly has been reported to be associated with oxidative stress. The therapy of kwashiorkor is still ineffective. In this pilot study, we tested the hypothesis that oral application of thiol-containing antioxidants increases glutathione status and is beneficial for the clinical recovery of kwashiorkor patients. The longitudinal clinical intervention study was carried out at St Joseph's Hospital, Jirapa, Ghana. Children with severe kwashiorkor were randomly assigned to either a standard treatment (ST) receiving a therapeutic protocol based on the recommendations of the WHO or to one of three study groups receiving in addition 2 x 600 mg reduced glutathione or 2 x 50 mg alpha-lipoic acid or 2 x 100 mg N-acetylcysteine per day. Patients were followed up clinically and biochemically for 20 days and compared with 37 healthy controls. Both glutathione and alpha-lipoic acid supplementation had positive effects on survival. Also, the blood glutathione concentrations correlated positively with survival rates. Furthermore, the initial skin lesions, glutathione and total protein concentrations were found to be strong predictors of survival. The data strongly suggest that a therapy restoring the antioxidative capacity by applying cysteine equivalents in the form of glutathione and/or alpha-lipoic acid is beneficial for biochemical and clinical recovery of kwashiorkor patients. PMID- 16259790 TI - Interaction and reactivity of urocanic acid towards peroxyl radicals. AB - The capacity of urocanic acid to interact with peroxyl radicals has been evaluated in several systems: oxidation in the presence of a free radical source (2,2'-azobis(2-amidinopropane; AAPH), protection of phycocyanin bleaching elicited by peroxyl radicals, and Cu(II)- and AAPH-promoted LDL oxidation. The results indicate that both isomers (cis and trans) are mild peroxyl radical scavengers. For example, trans-urocanic acid is nearly 400 times less efficient than Trolox in the protection of the peroxyl radical promoted bleaching of phycocyanin. Regarding the removal of urocanic acid by peroxyl radicals, nearly 100 muM trans-urocanic acid is required to trap half of the produced radicals under the employed conditions (10 mM AAPH, 37 degrees C). Competitive experiments show that the cis-isomer traps peroxyl radicals 30% less efficiently than the trans-isomer. Given the high concentrations that trans-urocanic acid reaches in skin, its capacity to trap peroxyl radicals could contribute to the protection of the tissue towards ROS-mediated processes. Furthermore, both isomers, and particularly the cis-isomer, protect LDL from Cu(II)-induced oxidation. PMID- 16259791 TI - Plasma non-esterified fatty acids: why are we not measuring them routinely? PMID- 16259792 TI - Caspase and calpain function in cell death: bridging the gap between apoptosis and necrosis. AB - Calpain and caspase are families of cysteine proteases that have important roles in the initiation, regulation and execution of cell death. The function of both groups of proteases in the progression of apoptotic and necrotic pathways is presented here in the context of a concise overview of regulated cell death. Many of the morphological differences between apoptotic and necrotic processes are thought to be as a consequence of the action of cysteine proteases. Recent studies suggest that caspase and calpain cascades are tightly interrelated and an appreciation of how these proteases cross-talk should enable a greater understanding of how the boundaries between apoptotic and necrotic cell death have become blurred. Furthermore, an assessment of the contribution that caspase and calpain make to human physiology and pathology is provided, with a description of how these proteases can be detected and quantified. Lastly, an evaluation is made of how caspase and calpain activation might be exploited diagnostically. PMID- 16259793 TI - Natriuretic peptides and the heart: current and future implications for clinical biochemistry. AB - The measurement of B-type natriuretic peptides in plasma is under intense promotion as a method of screening for heart failure. This article provides a historical context for this contention, and attempts to highlight what practical problems may be encountered in establishing a screening service from a clinical biochemistry standpoint. B-type natriuretic peptide measurements may also prove, in future, to have a significant role in the objective monitoring of treatment for heart failure, and to be a valuable prognostic indicator in patients suffering from acute coronary syndrome. PMID- 16259794 TI - Detection assessment and monitoring of hepatic fibrosis: biochemistry or biopsy? AB - Hepatic fibrosis is an important consequence of inflammatory disorders affecting the liver, and ultimately progresses to cirrhosis. Here we explore methods for the detection and monitoring of hepatic fibrosis, particularly in hepatitis C, alcoholic liver disease, non-alcoholic fatty liver disease and during methotrexate therapy, in all of which progressive fibrosis can develop over a number of years in a minority of patients. Liver biopsy currently remains the gold standard to assess fibrosis. However, it has several limitations, including manpower issues, cost, risk of patient injury, including mortality and morbidity, observer variability and sampling variation. Several non-invasive diagnostic tests for fibrosis and cirrhosis have therefore been evaluated. The usefulness of a laboratory test for screening for a pathological abnormality such as fibrosis is critically dependent on the prevalence of the pathology in the population under investigation. When the prevalence is expected to be low, screening tests should have a high negative predictive value so that large numbers of patients can be spared the next diagnostic step, namely liver biopsy. For the moment, clinical chemistry laboratories should offer the aspartate aminotransferase alanine aminotransferase ratio, AST/platelet ratio and the Rosenberg fibrosis index as part of their routine service for monitoring the development of hepatic fibrosis. PMID- 16259795 TI - Erythrocyte zinc in hospital patients. AB - BACKGROUND: Abnormal thyroid function tests (serum thyrotropin [TSH], free thyroxine [T(4)] and free triiodothyronine [T(3)]) are frequently seen in hospitalized patients. Assessment of thyroid function in these patients is difficult. It has been suggested that acutely ill patients may be hypothyroid at the tissue level. Erythrocyte zinc (EZn) has been shown to be increased in hypothyroidism. The aim of this study was to examine EZn as an index of thyroid status of hospital patients. METHODS: In order to assess the thyroid status at tissue level, we measured EZn in 26 healthy subjects, 39 critically ill patients and 19 hospitalized geriatric patients. EZn was measured in young cells, as the effect of illness is likely to be seen in the newly formed cells. RESULT: TSH and free T(3) were lower in critically ill patients and serum free T(3) was lower in geriatric patients. EZn in young cells was higher in both patient groups (by 13% and 23%, respectively). EZn in old cells was also higher in the geriatric group. CONCLUSION: We conclude that EZn is higher in hospitalized patients, suggesting that these patients may be hypothyroid at the tissue level. PMID- 16259796 TI - Potential pitfalls in intraoperative parathyroid hormone measurements during parathyroid surgery. AB - BACKGROUND: The outcome of parathyroid surgery is often not clear for at least 24 h after the operation. A frozen section does not always distinguish between an adenoma and hyperplasia. Minimally invasive surgical techniques are being refined, so the need for perioperative assurance about the completeness of surgery has increased. The value of intraoperative parathyroid hormone (PTH) measurements in 26 surgical cases undergoing parathyroidectomy has been evaluated. METHODS: Twenty-one patients were diagnosed as having primary hyperparathyroidism, including two patients with multiple endocrine neoplasia type I (MEN I). Five patients had tertiary hyperparathyroidism, including one patient with X-linked hypophosphataemia and four with renal hyperparathyroidism (RHPT). Blood samples were taken at the onset of surgery, at the time of tumour resection and at 5-min intervals following removal of the tumour. PTH was measured using a PTH Turbo assay on the DPC Immulite analyser. RESULTS: Current practice suggests that the PTH concentration should fall to less than 50% of the pre-incision value or to less than 50% of the level at the time of tumour resection (time equals zero). PTH levels were therefore monitored at 5-min intervals following removal of the tumour. In most of the case studies PTH followed the suggested pattern, but not when further exploration was warranted to determine if another adenoma was present. In some cases the PTH levels fell by the appropriate margin to deem the procedure a success but at 10 min post-gland excision the PTH began to rise again. Further exploration was required to confirm the continued source of PTH. CONCLUSION: We recommend that intraoperative PTH measurements continue until at least 15 min post-gland removal in cases of suspected single-gland disease. A decline in PTH concentration to at least 50% of the pre-incision or time of gland resection levels should be observed. If the PTH remains elevated or rises again after an appropriate decrease in levels, then multigland disease or ectopic sources should be considered. Caution is recommended in interpreting intraoperative PTH measurements to ensure complete success of the surgical procedure. PMID- 16259797 TI - The influence of genetic and environmental factors on plasma homocysteine concentrations in a population at high risk for coronary artery disease. AB - BACKGROUND: Elevated plasma total homocysteine (tHcy) predisposes to vascular disease and results from interactions between genetic and nutritional factors. MTHFR C(677)T increases tHcy in association with low folate. CBS 844ins68 lowers tHcy and negates the raising effect of MTHFR C(677)T in healthy subjects, but it is unclear if this is the case in subjects at high risk of vascular disease. This study examines the effect on plasma tHcy of interactions between these polymorphisms in an at-risk group. METHODS: Blood samples were collected from 376 subjects at increased risk of coronary artery disease. Plasma tHcy and vitamin B(6) were measured by HPLC and red cell folate and serum vitamin B(12) were measured by immuno-luminometric assay. MTHFR C(677)T and CBS 844ins68 status was established by standard PCR techniques. RESULTS: MTHFR TT predisposed to hyperhomocysteinaemia; this was increased in the presence of low folate (P<0.05) and vitamin B(12) (P<0.01). An inverse relationship was found between tHcy and folate (r=-0.42, P<0.0001), vitamin B(12) (r=-0.26, P<0.0005) and vitamin B(6) (r=-0.25, P<0.01). There was no interaction between plasma tHcy, vitamins or MTHFR C(677)T and CBS 844ins68. DISCUSSION: In this population at high risk of coronary artery disease, plasma tHcy was determined by vitamin status. This was exacerbated by the MTHFR C(677)T mutation. CBS 844ins68 did not influence tHcy and did not negate the tHcy-raising effect of MTHFR C(677)T. PMID- 16259798 TI - Do we need lipid clinics? Shifting the balance between secondary and primary care. AB - BACKGROUND: Increasing activity in cholesterol lowering is placing increasing demands on lipid clinics to be able to cope with the increase in demand. A combination of interventions has been used to improve laboratory testing, focus interpretative results and provide educational and advisory facilities for general practitioners in order to increase shared care of many potential clinic patients. METHODS: Retrospective study of clinic waiting times, results in patients managed on a shared care basis, and overall clinical activity over a three-and-a-half-year intervention period between March 2001 and August 2004 in a single-consultant, single-centre secondary care clinic serving three primary care trusts, covering a population of approximately 270,000 people. The interventions involved a change to the laboratory request form, outreach educational meetings and promotion of use of "written advice only" for certain patients as an alternative to direct clinic referral. The main outcome measures were percentages of patients followed up in primary care, change in cholesterol and triglyceride results after advice, and clinic waiting time. RESULTS: A total of 520 patients were referred over three and a half years, either to be seen or for written advice only. About 291 of these were handled by advisory letters. In all, 90% of these patients were already receiving or had received lipid-lowering therapy at the time of referral. 98% of patients were followed up by their general practice post-advice. Cholesterol and triglyceride concentrations fell by 23% and 39%, respectively, post-advice. Waiting times fell from a peak of 35 weeks before the interventions to an average of three weeks after. This fall has been maintained over the three-and-a-half years the intervention has run. CONCLUSIONS: The interventions have resulted in a large fall in clinic waiting times, improvement in lipid results, and high rate of general practice follow-up, all of which have been sustained in the long term. PMID- 16259799 TI - Development of a method for the measurement of dehydroepiandrosterone sulphate by liquid chromatography-tandem mass spectrometry. AB - BACKGROUND: Dehydroepiandrosterone sulphate (DHEAS) is a steroid that is increasingly being recognized as a potential drug of abuse in many countries. This is due to its reputation as a hormone that may be able to retard the ageing process. The measurement of DHEAS is useful in the diagnosis of medical conditions such as congenital adrenal hyperplasia and polycystic ovary syndrome. Thus, a liquid chromatography-tandem mass spectrometry method has been developed to determine DHEAS concentrations in human serum. METHOD: The chromatography was performed using a Waters 2795 Alliance HT LC system coupled to a Mercury Fusion RP column fitted with a SecurityGuard column. RESULTS: DHEAS and the internal standard, deuterated DHEAS, both had a retention time of 1.5 min. The transition determined by the Micromass Quattro tandem mass spectrometer for DHEAS was m/z 367.3>4 96.7 and for the internal standard m/z 369.3>96.6. The method was linear up to 20 micromol/L; the lower limit of detection and the lower limit of quantitation were both 1 micromol/L. The intra- and interassay imprecision were <11% over a concentration range of 1-18 micromol/L for the in-house quality control and <12% for the intra- and interassay imprecision for the Bio-Rad Lyphocheck QC. CONCLUSION: The measurement of DHEAS by liquid chromatography tandem mass spectrometry is robust and has a simple sample preparation procedure with a rapid cycle time of only 4 min. PMID- 16259800 TI - Interference of ascorbic acid with chemical analytes. AB - BACKGROUND: Ascorbic acid can interfere with methodologies involving redox reactions, while comprehensive studies on main chemistry analysers have not been reported. We therefore attempted to determine the interference of ascorbic acid with analytes on the Beckman Synchron LX20. METHODS: Various concentrations of ascorbic acid were added to serum, and the serum analytes were measured on the LX20. RESULTS: With a serum ascorbic acid concentration of 12.0 mmol/L, the values for sodium, potassium, calcium and creatinine increased by 43%, 58%, 103% and 26%, respectively (P<0.01). With a serum ascorbic acid concentration of 12.0 mmol/L, the values for chloride, total bilirubin and uric acid decreased by 33%, 62% and 83%, respectively (P<0.01), and were undetectable for total cholesterol, triglyceride, ammonia and lactate. There was no definite influence of ascorbic acid on analytical values for total CO(2), urea, glucose, phosphate, total protein, albumin, amylase, creatine kinase, creatine kinase-MB, aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, total iron, unbound iron-binding capacity or magnesium. CONCLUSIONS: Ascorbic acid causes a false increase in sodium, potassium, calcium and creatinine results and a false decrease in chloride, total bilirubin, uric acid, total cholesterol, triglyceride, ammonia and lactate results. PMID- 16259804 TI - Injectable contraceptives: a worldwide perspective. PMID- 16259806 TI - Depo-Provera and bone density. PMID- 16259808 TI - Depot medroxyprogesterone and bone mineral density. AB - OBJECTIVE: To investigate depot medroxyprogesterone (DMPA)-associated bone loss in a general practice setting. DESIGN: Forty-eight patients from a single practice who had used DMPA for contraception for more than 2 years. All patients had a serum oestradiol and if the serum level was <52 pmol/l or >52 pmol/l with menopausal symptoms, bone mineral densitometry (BMD) measurements were made at the lumbar spine (LS) and femoral neck (FN) using dual-energy x-ray absorptiometry (DEXA). Thirty-two patients had bone densitometry, of whom 27 had a serum oestradiol <52 pmol/l and five >52 pmol/l associated with menopausal symptoms. Of the remaining 16 patients, nine patients had a serum oestradiol <52 pmol/l but did not have a BMD as they moved away (five women) or switched to another contraceptive (four women). RESULTS: BMD results showed a significantly reduced bone mass at both sites with mean Z score LS -0.84 (95% CI -1.17 to 0.52) and FN -0.32 (95% CI -0.62 to -0.02). Eighteen women (56% of 32 women) had either osteopenia (15 cases) (T score < -1.0) or osteoporosis (three cases) (T score < -2.5) at the LS. There were trends to an association of a family history of height loss or tobacco smoking (current or past) for LS and FN Z scores that did not quite achieve significance. There was also a trend to lower body weight in those with a possible family history of osteoporosis or who were smokers and an inverse correlation of weight with BMD at the FN (p < 0.05) and a non significant inverse correlation at the LS. CONCLUSION: The present results demonstrate that a low bone mass should be considered in patients with prolonged DMPA usage especially if they have risk factors for osteoporosis. PMID- 16259809 TI - Questionnaire survey of women aged 56-59 years: consultations in general practice, use of hormone replacement therapy (HRT) and participation in screening programmes. AB - OBJECTIVES: To ascertain attitudes to consultations in general practice and the gender of the doctor. To determine whether women participate in the National Health Service (NHS) screening programmes available to them and their use of hormone replacement therapy (HRT). DESIGN: Postal questionnaire survey to women aged 56-59 years. SETTING: The Honiton Group Practice, East Devon, UK. PARTICIPANTS: Two hundred and ninety-four of the 334 eligible women returned the questionnaire (response rate 88%). MAIN OUTCOME MEASURES: Women's embarrassment on consulting general practitioners (GPs) and preference for a female doctor. Up to-date cervical smears and mammography screening in eligible women and use of HRT. RESULTS: A large majority of women (248/294; 84.4%) were not embarrassed when consulting a GP and did not express a preference to see a woman GP for any problem (240/294; 81.6%). However 123/294 (41.8%) did express a preference to see a woman doctor for 'women's problems'. Sixty-five women had had a hysterectomy, and 218 of the remaining 229 (95.2%) had had a smear in the past 5 years. For breast screening, 267/294 (90.8%) had undergone mammography in the previous 3 years. Ninety-three women (32%) were currently on HRT, and on record searching a total of 146 (49.7%) had at some time been HRT users. CONCLUSIONS: Embarrassment does not appear to be a major problem for consultations related to female health. A well-organised general practice with motivated patients can achieve very high uptakes of both cervical and breast screening programmes. Interest in HRT has also risen, with half the women aged 56-59 years having used systemic HRT. PMID- 16259810 TI - Sexual health work in a playground: lessons learnt from the evaluation of a small scale sexual health project. AB - OBJECTIVE: To evaluate a sexual health outreach programme in an adventure playground. DESIGN: A qualitative approach to evaluation is appropriate for a sexual health project where outcomes are unpredictable, difficult to define in numerical terms and the small scale of the intervention means that an impact on indices such as teenage pregnancy rates is unlikely. The data collection techniques used were the outreach worker's diary, questionnaires and interviews with youth workers, and a focus group with young people. RESULTS: Work in young people's leisure space requires flexible working in disrupted environments and a responsive approach to diverse individual needs. The outreach worker successfully engaged boys and girls in sexual health discussions. As a result of the project, youth workers' confidence in discussing sexual health issues and ability to refer to the clinic increased. CONCLUSION: Clinical staff providing a sexual health outreach service for young people offer a knowledge of sexual health and local clinical services and an opportunity to build positive relationships between clinics and local community organisations. The relationships help youth workers to raise sexual health issues with young people and to refer problems appropriately. Clinical staff doing outreach work may require additional skills to work in young people's leisure space, for example, the ability to negotiate the content of sessions and to tolerate disruption. A qualitative approach to evaluation effectively documents a range of project outcomes, including those which were unforeseen and which could not have been documented through the collection of numerical data. It also provides a detailed description of the process of project implementation, which aids replication. PMID- 16259811 TI - Anonymity and confidentiality: rural teenagers' concerns when accessing sexual health services. AB - CONTEXT: While confidentiality is recognised as a key aspect of successful health services aimed at young people, most research has looked at the concerns of those in urban centres. This paper reports on qualitative and quantitative data collected from general practitioners (GPs) and young people in a rural health district. OBJECTIVE: To assess the concerns of rural teenagers regarding anonymity and confidentiality when accessing sexual health services. DESIGN: The views of teenagers about using health services for issues of sexual health were sought through an in-school survey of 311 Year 9 and 119 Year 11 students. In addition, 18 single-sex focus groups discussions were conducted in North and East Devon. All GPs in the district were asked to complete a questionnaire. RESULTS: These reveal that the particular concerns of young people from small communities are more to do with the difficulties of remaining anonymous, which are related to visibility and lack of privacy in small communities. These problems were more pervasive among rural young people than those concerns more usually reported about confidential consultations. PMID- 16259812 TI - Prevention of migraine in the pill-free interval of combined oral contraceptives: a double-blind, placebo-controlled pilot study using natural oestrogen supplements. AB - CONTEXT: Migraine in the pill-free interval of combined oral contraceptives is reported by many women, but there is little published information on possible mechanisms and treatments. OBJECTIVE: To determine whether the use of natural oestrogen patches affected the occurrence and severity of migraine during the pill-free interval. DESIGN: A double-blind, placebo-controlled, randomised, crossover study. SETTING: The City of London Migraine Clinic. PARTICIPANTS: Fourteen women with migraine during the pill-free interval. INTERVENTIONS: 50 microg oestradiol patches (Evorel) used during the pill-free interval for two cycles versus placebo for two cycles (total four cycles). MAIN OUTCOME MEASURES: Number of pill-free intervals (zero, one or two) during which migraine occurred; number of days of migraine; severity of migraine; number of days of migraine accompanied by nausea, vomiting and/or photophobia. RESULTS: Complete data were available for 12 women and for two cycles for one woman. Use of 50 microg oestrogen patches during the pill-free interval showed a trend towards reducing the frequency and severity of migraine. DISCUSSION: These results were not as good as expected. However, we had originally aimed for 20 eligible women to participate in the trial, but only 14 were recruited and only 12 completed the study with full data for analysis. CONCLUSION: The results of this pilot study suggest that use of 50 microg oestrogen patches during the pill-free interval may reduce the frequency and severity of migraine at that time. This study should be repeated with larger numbers of women and a higher dose of oestrogen. PMID- 16259813 TI - Contraception and unexplained abdominal pain. AB - OBJECTIVE: To test the hypothesis that hospital referral for unexplained abdominal pain might occur less frequently in oral contraceptive (OC) users and more frequently in intrauterine device (IUD) users than in other women. DESIGN: Prospective cohort study of 17 032 women using different methods of contraception [the Oxford-Family Planning Association (FPA) contraceptive study]. OUTCOME MEASURE: Referral to hospital for unexplained abdominal pain (including pelvic pain) coded to ICD rubric 785.5 in the 8th Revision of the ICD. RESULTS: Referral was decreased by 13% (95% CI 0%-25%) in current or recent users of OCs in comparison with non-users, a difference that just reached statistical significance. No significant difference was found in the corresponding analysis for IUD users. Past users of OCs had much the same risk of referral as non-users, but the risk of referral in past users of an IUD was increased at all intervals examined, varying from 17% (95% CI -14%-55%) 13-24 months after discontinuation to 56% (95% CI 15%-107%) 73-96 months after discontinuation. Referral was 29% (95% CI 11%-50%) higher in smokers of 15 or more cigarettes a day than in non smokers. CONCLUSION: Referral to hospital for unexplained abdominal pain may be slightly reduced in current or recent OC users and moderately increased in past IUD users. The types of IUD used most commonly in the Oxford-FPA study are, however, no longer in current use today. Hospital referral for unexplained abdominal pain seems to be moderately increased in heavy cigarette smokers. PMID- 16259814 TI - Female sterilisations with Filshie clips: what is the risk failure? A retrospective survey of 30,000 applications. AB - OBJECTIVE: To assess the failure rate of Filshie clip sterilisations in Victoria, Australia. DESIGN: Retrospective study between 1994 and 1998. SETTING: All specialist gynaecologists practising in the state of Victoria, Australia. INTERVENTIONS: Questionnaires (up to three) followed by telephone calls if necessary. MAIN OUTCOME MEASURES: A response rate of 276/277 (99.6%) was obtained. All pregnancies were recorded. RESULTS: Seventy-three failures were notified from an estimated 30,000 Filshie clip sterilisations carried out in the study period. CONCLUSIONS: The estimated failure rate of Filshie clip sterilisations in the state of Victoria carried out by specialist gynaecologists was between 2 and 3 per 1000 sterilisation operations. PMID- 16259815 TI - Pilot study to assess the presence of Chlamydia trachomatis in urine from 18-30 year-old males using EIA/IF and PCR. AB - CONTEXT: To increase detection, urine samples from young males could be opportunistically tested for Chlamydia trachomatis. OBJECTIVE: To determine C. trachomatis prevalence in urine, optimum specimen and compare sensitivity/feasibility of routine use of different testing methods. DESIGN: Group A, 'sterile' pyuria samples June 1998-January 1999, tested by enzyme immunoassay (EIA) and, if reactive, by immunofluorescence (IF). Subsequently batch-tested by polymerase chain reaction (PCR). Group B, consecutive urine samples October 1998-January 1999; batch-tested by PCR. SETTING: Microbiology laboratory. SAMPLES: From males aged 18-30 years; group A = 71, group B = 83. MAIN OUTCOME MEASURES: Chlamydia trachomatis positive if EIA- and IF- or PCR positive. RESULTS: Group A: 12 EIA/IF-positive; 9/12 and 15 EIA-negative samples PCR-positive. Group B: 11 PCR-positive; 8/11 showed 'sterile' pyuria. CONCLUSIONS: Opportunistic testing of urine from young men shows a significant number of C. trachomatis infections. 'Sterile' pyuria samples are optimal. EIA/IF are less sensitive than PCR but can be routinely performed and detect a significant proportion of cases. PMID- 16259816 TI - Revisiting a pilot survey involving contraception and teenage pregnancy in Ayrshire and Arran. AB - CONTEXT: How to respond to the challenge of reducing unplanned pregnancy rates in Ayrshire and Arran. OBJECTIVES: (1) To improve understanding of the educational needs of 11-15-year-olds regarding contraception which could be used to inform planning of future sex education in schools in Ayrshire and Arran. (2) To put this in context by reviewing contraceptive usage amongst 14-16-year-old pregnant teenagers. DESIGN: (1) A questionnaire given to 11-15-year-olds during a sex education class. (2) A retrospective study of pregnant 14-16-year-olds. PARTICIPANTS: (1) School pupils aged 11-15 from an area of mixed social background (n = 80). (2) Pregnant 14-16-year-olds presenting at Ayrshire Central Maternity Hospital between September 1997 and March 1998 (n = 74). RESULTS: (1) Ninety-nine percent of the teenagers said they would use contraception. Seventy three percent of girls, but only 52% of boys, were aware of the services available. Ninety percent of girls knew about emergency contraception, but only 59% of boys. Thirty-three percent thought they received too little sex education at school. Thirty-two percent received no sex education from home, and 10% 'too little'. More information about sexually-related diseases, safe sex and having a baby was requested. (2) The study of pregnant teenagers showed that 69% went ahead with their pregnancies, that 71% were not habitually using contraception and only 4% were using contraception at the time of conception. DISCUSSION: Teenagers said they would use contraception. However, boys were not aware of local services giving advice and help to young people, nor about emergency contraception. Teenagers felt they had no or 'too little' sex education at home. They felt too young to become a parent and be 'tied down'. CONCLUSIONS: (1) Teenagers at school want information about a wider range of sexual issues. (2) There is a gap between learning about contraception and its practical use, including emergency contraception. (3) Discussion and reinforcement of sex education at home may not exist for many teenagers. (4) Teenagers at school felt that they were not ready to be parents as this would restrict their social lives and disrupt their education. (5) It is recognised that young men are less well informed than young women are, and this could impact on safe sex. PMID- 16259820 TI - Prosecution of rape and sexual assault. PMID- 16259822 TI - The problem of rape and supporting victims. PMID- 16259821 TI - Rape: the victim's perspective. PMID- 16259823 TI - One in four: providing emotional support to female victims of domestic violence. PMID- 16259824 TI - Sexual abuse and learning disability. PMID- 16259825 TI - Drug-facilitated rape. PMID- 16259827 TI - Assisting and advising complainants of sexual assault in the family planning setting. PMID- 16259829 TI - Uterine balloon therapy for menorrhagia: a feasibility study of its use in the community setting. AB - OBJECTIVES: To determine whether uterine balloon therapy (UBT) for menorrhagia can be performed safely in the community setting, obviating the need for hospital admission or general anaesthesia. DESIGN: Prospective case studies of 20 women undergoing Thermachoice endometrial ablation for menorrhagia. SETTING: Glasgow Centre for Family Planning and Reproductive Health Care, Greater Glasgow Primary Care NHS Trust, Glasgow, UK. PARTICIPANTS: Twenty women with menorrhagia unresponsive to medical therapy. MAIN OUTCOME MEASURES: Pain levels experienced by women during the procedure, measured by visual analogue scores and analgesia requirements postoperatively. RESULTS: Pain scores were in the range 0.1-6.6 (median 1.1) for outpatient hysteroscopy, compared to 0.1-9.8 (median 4.0) for uterine balloon therapy. No procedure was abandoned due to pain. CONCLUSION: UBT performed under local anaesthetic is tolerated well by patients. It is an effective treatment for menorrhagia, which is safe and easy to perform in the community setting. PMID- 16259830 TI - Atrophic vulvovaginitis in women aged 50 years and above, both current and non users of hormone replacement therapy, attending a genitourinary medicine clinic. AB - INTRODUCTION: In women aged 50 years and over attending a genitourinary medicine (GUM) department the commonest presentation was with symptoms of vulval soreness, irritation or dryness. Atrophic vulvovaginitis (AVV) was the commonest diagnosis made. This study was undertaken to determine if the presence of AVV was related to the use, or not, of systemic hormone replacement therapy (HRT). METHOD: A prospective study was made of all women aged 50 years and over attending a GUM department over a 3-month period. RESULTS: Of the 124 women seen in this age group, 60 (40%) had AVV and 28 (23%) had vaginal candidiasis. No difference was found in current users or non-users of HRT. CONCLUSION: Accurate diagnosis is essential in women with vulval symptoms to ensure that appropriate therapy is given. In the present study symptomatic women aged 50 years and over were more likely to have AVV than candidiasis irrespective of their use of systemic HRT. PMID- 16259831 TI - The evolution of the Marie Stopes electrocautery no-scalpel vasectomy procedure. AB - OBJECTIVE: To review the evolution of the procedure, the reoperation rate and efficacy data for vasectomies performed in Marie Stopes centres during the periods 1990-1994 and 1995-1999. DESIGN: Retrospective review of re-operation rates and primary and secondary failures during the periods 1990-1994 and 1995 1999. SETTING: Marie Stopes vasectomy centres in the UK. PARTICIPANTS: A total of 41 123 men undergoing vasectomy. RESULTS: The re-operation rate for the period 1990-1994 was 0.7% and fell to 0.46% for 1995-1999. The reported pregnancy rate fell from 1 in 1429 procedures for the period 1990-1994 to 1 in 2804 for 1995 1999. CONCLUSION: The results show that vasectomy has had a low failure rate well below that of other methods of birth control. The outcome data continue to improve over time with the evolution of improved techniques and surgical expertise. PMID- 16259833 TI - A study to assess the prevalence of chronic testicular pain in post-vasectomy men compared to non-vasectomised men. AB - OBJECTIVE: To assess the prevalence of chronic post-vasectomy testicular pain (CPTP) compared to the prevalence of chronic testicular pain in a control population of non-vasectomised men. METHODS: A retrospective postal study of 198 men who had a vasectomy more than 3 years previously at the Palatine Centre to determine the incidence of CPTP, of whom 101 (51%) replied (mean age 40.4 years, range 29-54 years, mean time since vasectomy 46.5 months). A control group of 102 men (mean age 40.2 years, range 28-55 years) who had not had a vasectomy were obtained via patients attending the community family planning clinics or associated general practitioner (GP) services in the same geographical area. RESULTS: Occasional non-troublesome discomfort was reported by 37/101 of the post vasectomy men compared to 21/102 of the control group. Occasional discomfort which was a nuisance was felt by 10/101 of the post-vasectomy group compared with 3/102 of the control group, and 6/101 in the post-vasectomy group reported pain severe enough to seek medical advice compared to only 2/102 controls. None of the post-vasectomy men regretted having had the operation. CONCLUSIONS: Only 6% of cases (compared to 2% of controls) experienced pain severe enough to seek medical advice in the 3-4-year post-operative period. However, there was a significant difference in the prevalence of occasional testicular discomfort between post vasectomy men and controls. Mostly these pains were not regarded as troublesome, however men attending for vasectomy counselling should be informed of the possibility of this morbidity. PMID- 16259834 TI - Countywide survey of the current practice of chlamydia detection in primary care. AB - BACKGROUND: The Chief Medical Officer's (CMO's) Advisory Group on Chlamydia trachomatis (chlamydia) published its report in 1998 and a national screening programme is anticipated. Meanwhile the Public Health Laboratory Service (PHLS) reports that the number of positive diagnoses of genital chlamydia continued to rise throughout the last decade. OBJECTIVES: To consider the current practice of Gloucestershire general practitioners (GPs) for detecting genital chlamydia infections, and based on the findings to help the development of local guidelines and sexual health service provision. DESIGN: Questionnaire survey. SETTING: Primary care groups within a single English county. METHODS: A questionnaire survey was sent to GPs. Responses were handled anonymously and pooled for analysis. MAIN OUTCOME MEASURES: Response rates from GPs on the types of tests used for detection of chlamydia infection, how frequently they test different patient groups for chlamydia infection and attitudes to contact tracing. RESULTS: In women, opportunistic screening is not routinely performed and the rate of diagnostic testing varies with presentation. The rate of testing is comparatively lower in men and over 50% of GPs refer symptomatic men directly to a genitourinary medicine (GUM) clinic. Fewer than 50% of respondents always or usually forward details to GUM for contact tracing. CONCLUSIONS: Opportunistic screening is performed by a proportion of GPs but there is scope for more screening in higher risk groups. There is scope to increase the number of tests for chlamydia infection in patients presenting with symptoms. A programme of training and education in conjunction with guidelines may have merit in unifying practice and making detection and management of chlamydia more effective. This will have financial and resource implications. PMID- 16259836 TI - Laboratory testing for Chlamydia trachomatis urogenital infections. PMID- 16259835 TI - Clinical management of chlamydia in general practice: a survey of reported practice. AB - BACKGROUND: Chlamydia trachomatis is the most common curable sexually transmitted infection (STI) in the UK. Prevalence of the infection in different health care settings has been found to be between 2% and 29% and diagnoses of the infection continue to rise significantly. It is estimated, however, that only 10% of all chlamydial infections are seen and treated in genitourinary medicine (GUM) clinics. Few data exist about the actual practice patterns of primary care providers in the management of chlamydia. The management of patients identified with the infection within this setting warrants attention. AIMS: To determine general practitioners' (GPs') and nurses' reported clinical practice in the management of genital chlamydial infection. To design a needs-based education guideline on the management of chlamydia in rural primary care. DESIGN: An anonymous, confidential, self-administered postal questionnaire was sent to all GPs and cervical cytology practice nurses in 84 practices in Dyfed Powys Health Authority (DPHA) area, Wales, UK. RESULTS: Following two mailings of the survey, responses were obtained from 75/84 practices (response rate 89%). A total of 130/307 GPs (response rate 42%) and 72/152 nurses (response rate 47%) in these practices responded. The data demonstrated that few tests for chlamydia are being carried out in general practice: 42% (55/130) of GPs and 54% (37/72) of nurses reported carrying out between one and four tests a month, and 35% (46/130) of GPs and 29% (21/72) of nurses reported performing less than one test a month. Very few positive results are being obtained. The majority of GPs (72%) and nurses (68%) routinely use the appropriate endocervical swab when testing for chlamydia, however 19% of GPs and 20% of nurses are using either an incorrect wooden-stemmed swab or are taking an inappropriate high vaginal swab. CONCLUSIONS: Few chlamydia tests are being performed in primary care in DPHA, Wales. Health professionals in general practice are in need of, and are in favour of, up-to-date training on all aspects of chlamydial identification and management in primary care. Attention needs to be focused upon resources and the paucity of GUM services and their access within the DPHA area. PMID- 16259837 TI - Caesarean section scar ectopic pregnancy following postcoital contraception. AB - This is believed to be the first reported case of an ectopic pregnancy following failed progestogen-only emergency contraception. The ectopic pregnancy was at the site of a previous caesarean section scar and was managed conservatively. PMID- 16259838 TI - Intrauterine device insertions: setting our standards. AB - OBJECTIVE: To determine if the documentation for intrauterine device (IUD) insertions was satisfactory and to agree minimum standards for practice. METHOD: A list was drawn up with minimum standards for documentation in the case notes. These were agreed at the clinical audit meeting. A retrospective analysis of case notes was done and a list completed for each IUD insertion. SETTING AND PARTICIPANTS: IUD fittings at all the Abacus clinics from 1 September 1999 to 31 October 1999. RESULTS: Of the 232 IUD insertions, the gold standard for documentation was met in 20%, however documentation of essential criteria was met 93% and all criteria 83% of the time. Each doctor received individual, confidential, comparative feedback. CONCLUSION: We were able to agree a minimum standard for documentation to enhance adequate counselling, safe insertion, communication with colleagues and risk management. PMID- 16259839 TI - Revisiting 'Fraser ruling competence' for under 16s in the UK. PMID- 16259840 TI - Measuring risk: how, when and why? PMID- 16259847 TI - Working in the transference: clinical and research perspectives. AB - The aim of this paper is to address a matter that has been noted, but not adequately confronted, in the psychotherapy research literature--that different psychoanalytic psychotherapists mean very different things by the expression transference interpretations. We explore the matter in three ways: firstly, by providing a brief overview of clinical perspectives on transference interpretations; secondly, by looking at examples of transference interpretations provided by researchers who have studied this form of clinical intervention; and thirdly, by reporting an empirical study of the quality, frequency, and distribution of transference interpretations employed by a psychoanalytic psychotherapist in the course of three routine psychodynamic assessment interviews in the National Health Service (NHS). We emphasize that the meaning of a clinical concept such as transference interpretation is grounded in a set of therapeutic principles and practices that are difficult to characterize by means of a succinct definition. It follows that we should be circumspect about generalizing from research on the therapeutic effectiveness of transference interpretations when what is meant may differ widely from study to study. PMID- 16259848 TI - A question of finding harmony: a grounded theory study of clinical psychologists' experience of addressing spiritual beliefs in therapy. AB - In spite of the increasing emphasis upon spirituality in the psychological literature, research continues to highlight concerns that issues relating to spirituality are regularly overlooked within a therapeutic setting. The aim of the current study was to develop an account of the way in which clinical psychologists understand and address spirituality within therapy. Semi-structured interviews were conducted with eight clinical psychologists. The transcripts were analysed using the qualitative methodology of grounded theory, which also informed the data collection process. Two core categories were developed and termed spirituality as an elusive concept and finding harmony with spiritual beliefs. The diversity of meaning surrounding spirituality and the relative lack of engagement was found to create difficulty for constructing spirituality coherently as a concept. When specifically considering methods for identifying and addressing spiritual beliefs, contrasting approaches were identified. This diversity in understanding and approach has implications for the process and outcome of therapy, which are discussed in detail. Recommendations are suggested for developing spirituality as a more coherent and accessible concept, both within professional dialogue and the therapeutic context. PMID- 16259849 TI - Variability in lay perceptions of depression: a vignette study. AB - This paper describes a study investigating lay perceptions of depression in terms of perceived severity. Students (N = 128) were presented with vignettes describing individuals with symptoms of depression based on DSM-IV. The descriptions were varied in terms of gender, social status, and a self-referent manner of communicating depressive symptomatology. Participants were asked to rate the degree to which vignette characters were thought to be depressed on a Likert-type scale. Results indicate that a non-self-referent style of communicating depressive symptoms by female vignette characters was seen as an indication of elevated levels of depression, and these findings are discussed with reference to the literature. PMID- 16259850 TI - Stability of attachment-related anxiety and avoidance and their relationships with the five-factor model and the psychobiological model of personality. AB - OBJECTIVES: The stability of attachment-related anxiety and avoidance and their relationship with influential personality models are issues worth further investigation. Studying discriminant validity with respect to aspects of personality deemed as prevalently heritable, such as temperament, is particularly important. Our aims were to examine the stability of attachment dimensions and to study their relationship with psychological distress, the Big Five factors, and temperament and character dimensions. METHODS: The Experiences in Close Relationships (ECR) questionnaire, the State-Trait Anxiety Inventory (STAI), the Zung depression scale (ZDS), the Temperament and Character Inventory (TCI-125), and the Big Five Questionnaire (BFQ) were administered to 222 undergraduate and graduate students. After 1 month, 115 participants completed again the ECR, the STAI, and the ZDS. RESULTS: Attachment-related anxiety was modestly correlated with depression and anxiety, while attachment-related avoidance was modestly correlated with depression. The test-retest reliability of ECR scores was high. Changes in attachment-related anxiety were not related to changes in depression or anxiety, and changes in attachment-related avoidance were modestly correlated with changes in depression. In separate multiple regression models also including gender and age, attachment-related anxiety was correlated with harm avoidance, reward dependence, low novelty seeking, low self-directedness and low cooperativeness; low energy/extraversion and low emotional stability; attachment related avoidance was correlated with low reward dependence and tended to be correlated with low self-directedness. CONCLUSIONS: These findings corroborate the reliability and the convergent and discriminant validity of the ECR, and support the notion that the attachment dimensions are only modestly related to psychological distress and are not redundant with constructs developed within influential personality models such as the five-factor model and the psychobiological model. PMID- 16259852 TI - Self-reported attachment styles and therapeutic orientation of therapists and their relationship with reported general alliance quality and problems in therapy. AB - The aims of this study were to explore the relationship between therapists' self reported attachment styles and therapeutic orientation with the self-reported general therapeutic alliance and therapist-reported problems in psychological therapy.A sample of 491 psychotherapists from differing therapeutic orientations responded to a postal questionnaire. The questionnaire contained standardized measures of therapeutic alliance quality, attachment behaviours, a checklist of problems in therapy, and a brief personality inventory.Therapist-reported attachment styles generally explained a significant additional proportion of the variance in alliance and problems in therapy, over and above variance explained by general personality variables. Self-reported secure attachment style was significantly positively correlated with therapist-reported general good alliance. Self-reported anxious attachment styles were significantly negatively correlated with good alliance, and significantly positively correlated with the number of therapist-reported problems in therapy. Therapeutic orientation independently predicted a small but significant amount of the variance in reported general alliance quality in addition to that explained by attachment behaviours. PMID- 16259851 TI - Clinical significance and patients' perceived change in four sessions of brief psychodynamic intervention: characteristics of early responders. AB - This study investigated Tingey, Lambert, Burlingame, and Hansen's (1996) extension of Jacobson, Follette and Revenstorf's (1984) proposal for assessing clinical significance. Seventy (N=70) outpatients with/without Cluster C personality disorders treated with a brief psychodynamic intervention (BDI) were included in the study. Results showed that 33% of patients demonstrated clinically significant change on the Global Severity Index. Patients who improved reported more perceived subjective change, greater satisfaction with the treatment, and greater improvement on the Social Adjustment Scale than patients who did not improve (60%) or deteriorated (7%). Further analyses showed that clinical significance achieved in a four session ultra-brief therapy is associated with patient characteristics such as co-morbid personality disorders, level of defensive functioning, and specific interpersonal problems. Results were maintained at 3 month and 6 month follow-ups. Findings are discussed in reference to Howard's suggestions on remoralization and remission. PMID- 16259853 TI - From soap opera to science: towards gaining access to the psychopaths who live amongst us. AB - This idiographic and essentially exploratory study examined the unique experiences of 20 women who had been victimized within the context of heterosexual relationships with a male partner who was rated as having the characteristics associated with psychopathy. An integrated approach, using questionnaires, and biographical and narrative data, was utilized. The experimental group consisted of 20 women who had partnered males rated by the women on the Hare P-SCAN (Hare & Herve, 1999) as having many or most of the features of psychopathy. As this was the first reported use of the Hare P-SCAN to provide partner ratings, norms for scorings on this assessment tool were obtained from ratings provided by a control group of a random sample of 100 women who had completed the Hare P-SCAN and provided ratings for their last male partner. Four two-tailed independent samples t tests were conducted to test for significant differences in scorings between the two groups. Following screening, semi structured interviews were conducted with the experimental group, that is, 20 women who had partnered males with psychopathic traits. The four independent samples t test showed that significant differences occurred between the ratings of male partners by women in the experimental group and ratings provided by the control group. Narrative data, generated from written accounts and interviews with women in the experimental group, was successful in clarifying the manner in which males with the personality traits which define psychopathy managed the heterosexual relationship and the nature and patterns of the abusive behaviours which occurred in this context. PMID- 16259854 TI - The evolutionary psychology of eating disorders: female competition for mates or for status? AB - The relationship between eating disorders and female intrasexual competition (ISC) was studied. More specifically, it was predicted that Female ISC for mates would be the strongest predictor of bulimia, and that, in contrast, Female ISC for status would be the strongest predictor of anorexia nervosa. A group of 202 undergraduate women, attending the University of Arizona, Tucson, USA, participated in this study. These respondents completed surveys that contained the following measures: the Female competition for mates scale, the Female competition for status scale, the General Competitiveness Scale, the Eating Disorders Inventory (EDI), and an additional measure specific to Anorexia. The internal consistencies of the measures were computed using Cronbach's alpha, and the measures were found to have adequate measurement reliability. Correlations were computed and a structural equation model was constructed for all the subscales within the measures. The resulting model demonstrated that ISC for mates was ultimately the driving factor that contributed to Female competition for status, General competitiveness, Perfectionism, Body dissatisfaction, Drive for thinness, and both Bulimia and Anorexia. Contrary to initial expectations, the results supported a mostly spurious causal relationship between Female competition for status and anorexia, with the only indirect causal effect being through the influence of perfectionism, which was uniquely on anorexia and not on bulimia. The role of perceived personal and Ideal partner mate value was also explored. Although they were strongly positively related to each other, these were shown to have nearly equal and opposite effects on body dissatisfaction. PMID- 16259855 TI - Is the time right for direct entry into a career in health and biomedical informatics? PMID- 16259856 TI - A real-time, mobile phone-based telemedicine system to support young adults with type 1 diabetes. AB - Telemedicine systems have been proposed as a means of supporting people with diabetes in the self-management of their condition. Requirements for monitoring parameters of care, including glycaemic control, extent of analysis and interpretation of data, patient-clinician contacts, and involvement of a multidisciplinary care team with effective communication, can be addressed by telemedicine systems. We describe the development and implementation of an innovative real-time telemedicine system based around transmission and feedback of data to and from a mobile phone. Proprietary Java-based programs were used to link a blood glucose meter to a mobile phone. In addition to immediate transmission of blood glucose data, information about insulin dose, eating patterns and physical exercise were collected. Immediate feedback to the phone included a colour histogram to draw attention to levels of control over glycaemia over the previous two weeks. Clinicians supporting patients had access to summary screens identifying users not testing, and those with levels of blood glucose outside pre-defined limits. More detailed graphical displays of data were used to provide data about control of insulin dose and the degree to which it was modified in response to diet and exercise. The system has been evaluated in a clinical trial conducted in secondary care and is now being adapted for use in a trial in primary care, which is designed to assess its effectiveness in providing integrated management for the patient, general practitioner and pharmacist. PMID- 16259857 TI - Designing and implementing an electronic health record system in primary care practice in sub-Saharan Africa: a case study from Cameroon. AB - OBJECTIVE: To review the key issues related to the design and implementation of an electronic health record (EHR) system in urban primary health care (PHC) practice in Cameroon. METHODOLOGY: The goal of the project was to assess EHR as a tool to improve providers' performance, quality and continuity of care, and the availability of data in PHC practice in Cameroon. A locally designed EHR system called MEDCAB was developed. The system was based on the International Classification for Primary Care (ICPC) and was designed taking into consideration the PHC practice environment in Cameroon. An original cohort of 14 users was involved in the experiment. RESULTS: Users generally showed good acceptance of the system. Monitoring the use of the system at the early stages of implementation was important to ensure immediate response to users' comments and requests. Some of the key issues identified during the development and implementation of the system were: user involvement, the choice of an appropriate terminology, pre-existing data collection culture and leadership issues. Some positive achievements brought about by the system included promotion of good medical practice and routine availability of consultation data. CONCLUSION: Strengthening the medical record in general, and the EHR in particular, could contribute to its position as a valuable source of information for healthcare delivery, public health and policy making in Cameroon. Challenges to adoption are huge and successful implementation for any specific setting will require a comprehensive modelling of the local medical practice, the choice of an appropriate terminology and a co-ordinated approach involving all stakeholders. PMID- 16259858 TI - The implementation of electronic services: planned or organic growth? AB - The literature on innovation suggests that projects are successful when rigorous project management is mixed judiciously with 'organic' development. This paper argues that organic growth can play a substantial role in the implementation of electronic services in healthcare settings. Evidence for organic growth is presented, based on a study of email use. Methods are presented for investigating email use in health service settings in the National Health Service (NHS) in Bradford, England. Geographical information systems (GIS) outputs and social network analyses are presented. The results demonstrate a fivefold increase in the use of email over a 13-month period, which is shown to be largely independent of the growth in the number of organisations using the network. They also demonstrate a marked increase in the complexity of the patterns of email use over the period. PMID- 16259859 TI - Pandora's electronic box: GPs reflect upon email communication with their patients. AB - BACKGROUND: Global access to information technology has increased dramatically in the past decade, with electronic health care changing medical practice. One example for general practitioners (GPs) is communication with patients via electronic mail (email). GPs face issues regarding e-communication with patients, including how and when it should it be used. OBJECTIVE: The study aims were to assess the extent that GPs communicate with patients by email and explore their attitudes to this mode of communication. METHODS: Design--telephone interview survey. Setting--primary care, largest urban and suburban area in New Zealand (NZ). Subjects--randomly selected GPs from the Auckland region. Main outcome measure--description of email use; analysis of issues by telephone survey. Data analysed using SPSS-12 and by thematic content analysis. RESULTS: At data saturation, 80 GPs had been interviewed. The majority (68%) had not used email with patients. Only 4% used it regularly. However, there was strong interest in this method. Perceived advantages were the ability to communicate at a distance and time convenient to both doctor and patient; communication where disability affected traditional methods; information-giving (for example, web links); passing on normal results. Identified problems involved inequity of access; linking of electronic data; security; unsuitability for some topics; medico-legal concerns; time; remuneration. CONCLUSION: Study sample closely mirrored current NZ GP population. Although few GPs emailed with patients, many might once barriers are addressed. GPs had a collective view of the appropriate boundaries for email communication, routine tasks and the transmission of information. GPs would encourage professional debate regarding guidelines for good practice, managing demand and remuneration. PMID- 16259860 TI - Development of functional requirements for electronic health communication: preliminary results from the ELIN project. AB - BACKGROUND: User participation is important for developing a functional requirements specification for electronic communication. General practitioners and practising specialists, however, often work in small practices without the resources to develop and present their requirements. It was necessary to find a method that could engage practising doctors in order to promote their needs related to electronic communication. MATERIALS AND METHODS: Qualitative research methods were used, starting a process to develop and study documents and collect data from meetings in project groups. Triangulation was used, in that the participants were organised into a panel of experts, a user group, a supplier group and an editorial committee. RESULTS: The panel of experts created a list of functional requirements for electronic communication in health care, consisting of 197 requirements, in addition to 67 requirements selected from an existing Norwegian standard for electronic patient records (EPRs). Elimination of paper copies sent in parallel with electronic messages, optimal workflow, a common electronic 'envelope' with directory services for units and end-users, and defined requirements for content with the possibility of decision support were the most important requirements. CONCLUSIONS: The results indicate that we have found a method of developing functional requirements which provides valid results both for practising doctors and for suppliers of EPR systems. PMID- 16259861 TI - Randomised studies in general practice: how to integrate the electronic patient record. AB - The 'randomised database study' strategy was first proposed in 1997, with the aim of combining the generalisability of observational database studies based on electronic patient records (EPRs) with the validity of randomised clinical trials (RCTs). The key feature was to randomly assign treatments and to use routine care data, as available in the observational database, for patient identification and follow-up. To our knowledge, however, the idea of the randomised database study has not been implemented yet. The conduct of a randomised study in an observational database requires adjustments to methods of medical information processing in the general practice. We developed a software system that facilitates the conduct of an RCT with observational databases based on EPRs. It identifies eligible subjects and presents them one by one to the physician once their EPR is accessed. The general practitioner can then start an interactive recruitment process; after completion, the computer randomises the patients. Follow-up is documented by normal routine care in the EPR. Although the randomised database study has many methodological advantages, it has never been tested. Our software system is meant as a tool to implement and facilitate evaluation of the randomised database approach. PMID- 16259862 TI - INRstar: computerised decision support software for anticoagulation management in primary care. AB - Computerised decision support software (CDSS) for anticoagulation management has become established practice in the UK, offering significant advantages for patients and clinicians over traditional methods of dose calculation. The New GMS Contract has been partly responsible for this shift of management from secondary to primary care, in which INRstar has been the market leader for many years. In September 2004, INRstar received the John Perry Prize, awarded by the PHCSG for excellence and innovation in medical applications of information technology. PMID- 16259863 TI - Links between systems in Accident & Emergency and primary care. AB - The hospital emergency department and other elements of rapid access primary care constitute an emergency care network. Integration aims to maximise the network's strengths and overcome its weaknesses. Taking the patient as a starting point, it is possible to envisage an objective data model that can operate at multiple levels within the network to describe its process efficiency and clinical effectiveness. Other means of integration are also identified. These contain significant subjective elements. In particular, the decision support system of NHSDirect has operated successfully to legitimise national and local intervention based on skill-mix, whereas its technical operation has been susceptible to human deviation from prescribed routine. As we scrutinise a system, we discover that it contains people who are doing things. Logical elements in the system turn out to be givers or recipients of deliberate and thoughtful care. Information systems in Accident & Emergency (A&E) and primary care can help accountable planners to measure and control aspects of the network's operation. Clinicians also need their systems to enable, rather than constrain, effective interactions. PMID- 16259864 TI - The National Alliance for Primary Care Informatics: an update. PMID- 16259865 TI - Minimally invasive surgery in prostate cancer: current and future perspectives. AB - Prostate cancer is a leading cause of death for men. Its prevalence increases substantially with advancing age, which coincides with increased incidence of comorbidities. The mainstay therapy for early prostate cancer remains radical surgery and radiotherapy. In the era of prostate-specific antigen and the newly emerging highly specific screening tests, more men are diagnosed earlier in their life and with confined disease. The application of minimally invasive approaches, such as the laparoscopic/telerobotic approach and focal treatments, is becoming more popular, yielding shorter hospital stays and faster recovery, with minimal complications. In part, this is possible due to groundbreaking advances in laparoscopic equipment, prostate imaging modalities, and gained urologic expertise in minimally invasive treatment. In this review we critically discuss the current knowledge and future perspectives of minimally invasive treatment in prostate cancer. PMID- 16259866 TI - Clinical trials involving vitamin D analogs in prostate cancer. AB - Vitamin D shows significant potential as a therapy for prostate cancer. However, its use in clinical trials has been hampered by its induction of hypercalcemia at serum concentrations required to suppress cancer cell proliferation. This has spurred the development of less calcemic analogs of vitamin D. In this article, we review the clinical trials and consider the future directions of the use of vitamin D and its analogs in the treatment or chemoprevention of prostate cancer. First, we summarize the epidemiological evidence leading to the hypothesis that vitamin D has anticancer activity. We then review the clinical trials using vitamin D analogs that involve patients with prostate cancer and conclude with a brief overview of our planned study with vitamin D5, [1alpha(OH)D5], which will begin shortly. Data for this review were identified by searches of PubMed, the Cochrane Library, Biosis, and references from relevant articles, using the search terms "vitamin D," "prostate cancer," "chemoprevention" and "vitamin D analog." Abstracts from recent international meetings were also reviewed but were only included when they were the only known reference to the clinical trial or the research mentioned. Only papers published in English were included. PMID- 16259867 TI - Immunoproliferative small intestinal disease: current concepts. AB - Immunoproliferative small intestinal disease is a distinctive lymphoproliferative disorder. Among these disorders, it is the only disease associated with a specific and characteristic abnormal protein, and also an identifiable, at least in some patients, early phase with a benign-looking histo-pathologic expression. Whether the disease at this stage is malignant or not is not known. Treatment of this early phase with antibiotics may cause remission in some patients. This observation is significant and raises the question of chemoprevention in lymphomas. In contrast to primary nonimmunoproliferative small intestinal lymphomas, in which the pathology in the intestine is usually focal and involving specific segments of the intestine and leaving the segments between the involved areas free of disease, the pathology in immunoproliferative small intestinal disease is diffuse, with a mucosal cellular infiltrate involving large segments of the intestine and sometimes the entire length of the intestine, thus producing malabsorption. Preliminary recent epidemiological data have shown a decrease in the incidence of this disease in endemic areas, and therefore environmental factors are suspected to play a major role in its pathogenesis. Additional research is indicated not only to understand this specific lymphoproliferative disorder but also to understand lymphomas in general. PMID- 16259868 TI - Prostate implant therapy: iodine-125 versus palladium-103. PMID- 16259869 TI - I-125 versus Pd-103 for low-risk prostate cancer: long-term morbidity outcomes from a prospective randomized multicenter controlled trial. AB - BACKGROUND: We tested the hypothesis that the shorter half-life of Pd-103 versus I-125 results in different late radiation-related morbidities following prostate brachytherapy. METHODS: As of June 14th, 2002, 352 of a planned total of 600 patients with 1997 American Joint Committee on Cancer (AJCC) clinical stage T1c T2a prostatic carcinoma (Gleason grade 2-6, PSA 4-10 ng/mL) had been randomized to implantation with I-125 (144 Gy, TG-43) or Pd-103 (125 Gy, NIST-99). Treatment related morbidity was monitored by questionnaires based on standard American Urologic Association (AUA) and Radiation Therapy Oncology Group (RTOG) criteria that were mailed at 1, 3, 6, 12, 18, and 24 months after implant. The use of alpha-blockers to relieve obstructive symptoms was not controlled for but was noted at each follow-up point. All patients reported here had a minimum follow-up of 2 years. Dosimetric parameters analyzed included the V100, which was defined as the percentage of the postimplant prostate volume covered by 100% of the prescription dose. Rectal doses were expressed as the R100, defined as the rectal volume (cc) that received at least 100% of the prescription dose. Statistical comparisons were by Student's unpaired t-test at specified follow-up times. RESULTS: The AUA scores peaked at the 1-month postimplant time point for both isotopes and gradually declined. The difference in AUA scores between patients who received I-125 versus those who received Pd-103 was greatest at 1 and 6 months following implantation. At 1 month, I-125 patients had a mean AUA score of 14.8 (+/-9.5) compared with 18.6 (+/-9.8) for the Pd-103 patients (P = 0.0009). By 6 months, mean AUA scores for the I-125 patients had decreased to 12.0 (+/ 9.1) compared with 9.9 (+/-8.7) for the Pd-103 patients (P = 0.04). The use of alpha-blockers was similar between groups at all time points. Radiation proctitis (persistent bleeding) occurred in 29 of 314 patients (9%). There was an overall trend toward more proctitis in I-125 patients (P = 0.21). However, only four of the 163 patients (2%) with an R100 below the recommended 1.0 cc developed bleeding, which did not differ between isotopes (P = 0.49). DISCUSSION: Patients treated with Pd-103 had more intense radiation prostatitis in the first month after implantation, but they recovered from their radiation-related symptoms sooner than I-125 patients, consistent with palladium's shorter half-life. The trend toward more proctitis in the I-125 patient group likely reflects their higher R100 values due to less rapid dose fall-off that can be overcome with judicious treatment planning and implant execution. PMID- 16259870 TI - Effects of breast-conserving therapy on lactation after pregnancy. AB - BACKGROUND: As the incidence of breast-conserving therapy in women of childbearing years increases, patient concerns regarding subsequent pregnancies and lactation have become more prevalent. There is a paucity of data regarding lactation outcomes in women who have undergone breast-conserving therapy and then sustained full-term pregnancies. Our objective was to evaluate lactation outcomes in patients with early-stage breast cancer treated with breast-conserving therapy. METHODS: We reviewed a database of over 3,000 patients treated from 1965 to 2003 to identify our cohort of premenopausal women who underwent breast conserving therapy and subsequently sustained full-term pregnancies. Lactation outcome parameters (breast swelling, ability to lactate, and volume of lactation in the treated and untreated breasts) were the main outcome measures. RESULTS: We identified 28 pregnancies in 21 patients. The median age at diagnosis was 32 years. One patient underwent bilateral breast treatment; therefore, a total of 22 breasts were irradiated. All patients interviewed reported little or no swelling of the treated breast during pregnancy. Of the patients studied, 4 (18.2%) elected pharmacological suppression of lactation. Of the remaining 18 breasts, lactation occurred in 10 (55.6%), did not occur in 7 (38.9%) and was unknown for 1 (5.5%). The volume was reported as significantly diminished in 80% of breasts treated. Lactation in the contralateral breast occurred in all patients who did not undergo pharmacological suppression. CONCLUSION: Patients can experience successful lactation in the contralateral, untreated breast after breast conserving therapy. In the treated breast, functional lactation is possible but is significantly diminished in the majority of patients. PMID- 16259871 TI - Immunohistochemical biomarkers in patients with early-onset breast carcinoma by tissue microarray. AB - Young women with breast cancer have a poor prognosis, and the role of biologic markers in young women is not well defined. We investigated the association of estrogen receptor, progesterone receptor, Bcl-2, HER-2/neu, p53, and Ki-67 with clinicopathologic features and outcome in young women with breast cancer. METHODS: A cohort of 103 patients with early-onset breast cancer treated with conservative surgery and radiotherapy were entered in this study. Age range was 25-45 years, and median follow-up was 8.7 years. Each of the paraffin-embedded specimens was immunologically stained for six biomarkers expression by a recently developed tissue microarray method. RESULTS: The 10-year overall breast relapse free and distant relapse-free survival rates were 82.7%, 84.6.4%, and 66.7%, respectively, with 14 local relapses and 26 distant metastases among the 103 patients evaluated. Positive expression of estrogen receptor, progesterone receptor, bcl-2, HER-2/neu, p53, and Ki-67 were 42.7%, 48.5%, 35.6%, 28.0%, 36.9%, and 39.7%, respectively. Tumor stage and nodal status were significantly associated with overall survival and distant metastasis-free rate in univariate and multivariate analysis. Progesterone receptor negativity and Ki-67 positivity were associated with distant metastasis. There was no statistically significant correlation between the six biomarkers and local relapse. CONCLUSIONS: Progesterone receptor, Ki-67, tumor stage, and nodal status were prognostic factors for distant failure in early-stage breast cancer in young patients. Further studies are needed to find other biologic markers associated with local failure in this group of patients. PMID- 16259872 TI - Pathological distribution of positive lymph nodes in patients with clinically and radiologically N0 oropharyngeal carcinoma: implications for IMRT treatment planning. AB - BACKGROUND: We identified involved lymph node groups in patients with oropharyngeal carcinoma and radiologically N0 disease, aiming to define the clinical target volume, and to assess the negative predictive value of CT neck and correlate it to the tumor site, stage, and grade. METHODS AND MATERIALS: Between 1988 and 2000, we evaluated 53 patients who satisfied all of the following criteria: 1) oropharyngeal carcinoma diagnosis; 2) NO stage based on CT; 3) no prior treatment; and 4) primary surgical resection including ipsi lateral neck dissection. The pathology reports were reviewed to identify the exact site of lymph node involvement. RESULTS: Twenty patients (37.7%) were found to have pathologically positive lymph nodes, yielding a negative predictive value of 62.3% for CT neck. Node levels II, III, and IV were the most commonly involved (26%, 17%, and 11% of all patients, respectively). Fifty percent of patients with T3 and T4 tumors had positive lymph nodes versus 20% of patients with T1 and T2 (P = 0.036). Tumor grade and site were insignificant (P > 0.05). DISCUSSION: Ipsilateral neck levels II-IV should be included during elective nodal irradiation in patients with N0-stage oropharyngeal carcinoma, regardless of the primary tumor site, stage, and grade. PMID- 16259873 TI - Interstitial lung disease in Japanese patients with non-small cell lung cancer receiving gefitinib: an analysis of risk factors and treatment outcomes in Okayama Lung Cancer Study Group. AB - Risk factors for the development of interstitial lung disease in patients with non-small cell lung cancer receiving gefitinib and the prognostic factors after interstitial lung disease development have not been established. The aim of this study was to retrospectively identify and evaluate these possible factors. PATIENTS AND METHODS: We reviewed the clinical records and radiographs of 365 consecutive patients with non-small cell lung cancer who received gefitinib in West Japan between 2000 and 2003. RESULTS: In total, 330 patients were eligible for interstitial lung disease evaluation, and 15 patients (4.5%) were finally confirmed to have developed interstitial lung disease by blinded expert review. Multivariate analysis revealed that preexisting pulmonary fibrosis, poor performance status, and prior thoracic irradiation were independent risk factors for interstitial lung disease, with odds ratios of 21.0 (95% confidence interval, 5.12-86.3, P < 0.0001), 9.70 (2.27-41.4, P = 0.001), and 4.33 (1.27-14.8, P = 0.019), respectively. Among the 15 patients who developed interstitial lung disease, eight have died of the condition. Short interval from the initiation of gefitinib treatment to the onset of interstitial lung disease, acute interstitial pneumonia pattern, and the presence of pre-existing pulmonary fibrosis were associated with poor prognosis. DISCUSSION: Our results suggest the importance of patient selection for gefitinib treatment based on interstitial lung disease risk factors in the Japanese population identified. PMID- 16259875 TI - Permanent prostate brachytherapy (PPB) as it concerns seed migration and extraprostatic seed placement. PMID- 16259874 TI - Refractory or relapsed Hodgkin's disease and non-Hodgkin's lymphoma: optimizing involved-field radiotherapy in transplant patients. AB - This study assessed efficacy, optimal dosage and timing, and toxicity of involved field radiotherapy used in conjunction with high-dose chemotherapy and stem cell transplantation for patients with refractory/relapsed Hodgkin's disease and non Hodgkin's lymphoma. METHODS AND MATERIALS: 306 patients with refractory or relapsed Hodgkin's disease and non-Hodgkin's lymphoma were analyzed. Forty-one patients underwent involved-field radiotherapy in conjunction with high-dose chemotherapy and bone marrow or peripheral stem cell transplantation. Thirty three patients received involved-field radiotherapy prior to stem cell transplantation directed at symptomatic and/or bulky sites; eight patients received involved-field radiotherapy after stem cell transplantation directed at sites of persistent disease. The other 265 patients with refractory/relapsed non Hodgkin's lymphoma and Hodgkin's disease received high-dose chemotherapy/stem cell transplantation, but not involved-field radiotherapy. Data were analyzed using Cox proportional hazards regression to determine the risk of death among patients treated with stem cell transplantation compared with that among patients treated with stem cell transplantation and involved-field radiotherapy. RESULTS: There were 124 deaths during the follow-up period, including 17% of the patients treated with involved-field radiotherapy and 44.2% of the patients receiving chemotherapy without involved-field radiotherapy. Multivariate analysis found that patients who did not receive involved-field radiotherapy were 2.09 times more likely to die during the follow-up period than patients who received involved-field radiotherapy (P = 0.066; adjusted for age, stem cell transplantation type, stage I/II vs stage III/IV, refractory vs relapsed, and Hodgkin's disease vs non-Hodgkin's lymphoma). When patients were treated with involved-field radiotherapy prior to stem cell transplantation, 27 (79.4%) of the 34 patients achieved local control; when involved-field radiotherapy followed stem cell transplantation, 6 (85.7%) of the 7 patients experienced local control. Timing of involved-field radiotherapy prior to or following stem cell transplantation did not affect patient survival. Five of the 41 patients treated with involved-field radiotherapy developed toxicity subsequent to treatment. All but one of these patients had been treated with doses greater than 30 Gy. CONCLUSIONS: Although of borderline significance in this small sample, results of this study suggest that patients who receive involved-field radiotherapy in conjunction with stem cell transplantation may have increased survival when compared with patients who do not receive involved-field radiotherapy. Further follow-up of this cohort is necessary to confirm these findings. PMID- 16259876 TI - Intra-abdominal pressure in the morbidly obese. AB - BACKGROUND: Recent data suggests that increased intra-abdominal pressure (IAP) is one factor associated with the morbidity of morbidly obese patients, who have a BMI>35 kg/m2. IAP has been proposed to be an abdominal compartment syndrome (ACS). This study investigated the characteristics of IAP in morbidly obese patients. METHODS: 45 morbidly obese patients (mean BMI55+/-2 kg/m2) had IAP measured using urinary bladder pressure. RESULTS: The mean IAP for the morbidly obese group was 12+/-0.8 cmH2O, increased when compared to controls (IAP=0+/-2 cmH2O). The IAP correlated to the sagittal abdominal diameter, an index of the degree of central obesity (r=+0.83, P<0.02); however, it did not correlate to basal insulin, body weight, or BMI. The end-expiratory IAP did not change when measured after the laparotomy incision was made, but IAP measured in the last 15 patients increased during the first 2 postoperative days. The IAP for patients with pressure-related morbidity (gastroesophageal reflux disease, hernia, stress incontinence, diabetes, hypertension, and venous insufficiency) was 12+/-1 cmH2O, while those without these morbidities had an IAP of 9+/-0.8 cmH2O. CONCLUSION: We conclude that IAP is increased in morbid obesity. This increased IAP is a function of central obesity and is associated with increased morbidity. The degree of IAP elevation correlates with increased co-morbidities. We also conclude that elevation in IAP in morbid obesity is not a true ACS but represents a direct mass effect of the visceral obesity. PMID- 16259877 TI - Magnetic resonance spectroscopy facilitates assessment of intramyocellular lipid changes: a preliminary short-term study following biliopancreatic diversion. AB - BACKGROUND: Intramyocellular content of lipid (IMCL) appears to be important in the pathogenesis of insulin resistance, and the improvement of insulin activity observed following biliopancreatic diversion (BPD) for obesity has to be related to reduction of IMCL. This study evaluates the possibility of detecting changes after BPD in IMCL by means of H1 magnetic resonance spectroscopy. METHODS: The investigation was carried out in obese patients undergoing BPD prior to and at 1 month following the operation. Insulin sensitivity was assessed according to the homeostatic model of assessment (HOMA), and IMCL was determined by the spectroscopy of the tibialis anterior muscle. RESULTS: At 1 month following BPD, an improvement in insulin action was observed in all subjects, and in nearly all cases the IMCL signal decreased. CONCLUSION: The data from this study indicate that magnetic resonance spectroscopy can detect IMCL changes following BPD and is fully suitable for longitudinal studies on muscle metabolic status. PMID- 16259878 TI - Gait cinematic analysis in morbidly obese patients. AB - BACKGROUND: Functional co-morbidities of excess body weight such as gait problems are never life-threatening like those associated with certain metabolic sequelae. Nevertheless, they may interfere with quality of life and also act as a mirror of muscle, bone and joint stress. In this prospective study, the goal was to document dynamic aspects of gait in severely obese subjects. METHODS: An outpatient population (age 47.2+/-12.9 years, 94.1% females, BMI 40.1+/-6.0 kg/m2, n=34) had their gait analyzed by an experienced physical therapist. Variables included speed, cadence, stride, support base and foot angle, which were compared to reference values for the Brazilian population. RESULTS: All variables were significantly lower in the obese patients, except for support base which was increased. Speed was 73.3+/-16.3 vs 130 cm/s, cadence was 1.4+/-0.2 vs 1.8 steps/s, stride was 106.8+/-13.1 vs 132.0 cm, and support was 12.5+/-3.5 vs 10.0 cm (P<0.05). CONCLUSIONS: 1) Widespread cinematic impairment was the rule in the studied population. 2) These findings are consistent with poor skeletal muscle performance, high metabolic expenditure and constant physical exhaustion. 3) Attention should be paid not only to the metabolic management but also to the physical rehabilitation required in cases of advanced obesity. PMID- 16259879 TI - Does nonarticlar tenderness change after bariatric surgery? AB - BACKGROUND: A previous study reported an association between obesity and increased tenderness. However, the effect of weight reduction on tenderness is not known. The aim of the study was to assess tenderness thresholds before and after bariatric surgery. METHODS: 42 obese women were evaluated for tenderness before and 6 months after bariatric surgery. A count of 18 tender points at 9 symmetrical sites was performed by thumb palpation. 13 point sites (9 tender point sites and 4 control sites) were further studied using a dolorimeter. RESULTS: The obese subjects displayed increased tenderness before surgery and its level did not change 6 months later. The mean tender point count before surgery was 8.8+/-3.9 and after surgery was 8.3+/-4.3 (P=0.500). The mean tenderness thresholds at nine tender points were 4.0+/-1.1 and 3.8+/-1.1, respectively (P=0.247). CONCLUSION: Nonarticular tenderness in obese women remained high after weight reduction. These findings are relevant to physicians taking care of obese patients. Further studies are needed to elucidate the relationship between weight reduction and pain thresholds. PMID- 16259880 TI - Utilization of intensive care resources in bariatric surgery. AB - BACKGROUND: Obese patients occasionally require either elective or emergency critical care services following bariatric surgery. We describe this subgroup of patients. METHODS: From July 1, 1991 to July 31, 2004, we performed 1,279 bariatric operations; 241 (19%) required admission to the surgical critical care service. We retrospectively reviewed medical records for gender, body mass index (BMI), age, whether the operation was initial or revisional, and whether critical care admission was elective or emergent. 3 complication clusters (thromboembolic, pulmonary, and anastomotic) were identified using discharge ICD-9 codes. The costs and length of stay of these subpopulations was calculated. RESULTS: Patients were on average 46+/-10 years old, with BMI 59+/-13. Critical care admission was emergent in 52.7% (n=127) of cases. Revisional cases did not differ from the initial cases in BMI (56.4 vs 59.2, P=0.42) and they were no more likely to require emergent critical care admission than initial cases (P=0.16). Revisional cases were hospitalized longer (27.2+/-25.6 vs 12.5+/-18.7 days, P=0.003); had higher total hospital costs (US$ 60,631+/-78,337 vs 27,697+/ 52,351, P=0.025); and were more likely to die from their complications (revisional surgery mortality 6.5% vs 1.9% for initial surgery [P=0.002]). CONCLUSIONS: An increasing number of surgical revisions will likely accompany the recent increase in popularity of bariatric surgery. In our experience, these patients require significant critical care services, and have longer, complicated, and more costly hospitalizations. PMID- 16259881 TI - Conservative management of anastomotic leaks after 557 open gastric bypasses. AB - BACKGROUND: One of the most serious complications after gastric bypass is an anastomotic leak. In a prospective surgical protocol for the management of this complication, the authors determined the incidence of anastomotic leaks. METHODS: From August 1999 to January 2005, 557 patients with morbid obesity were submitted to laparotomic resectional gastric bypass. In all patients a left drain was placed during surgery. All patients had a radiological study with liquid barium sulphate on the 5th postoperative day. After the occurrence of an anastomotic leak, the daily output of the leak was carefully measured. RESULTS: 12 patients developed an anastomotic leak at the gastrojejunostomy. All were managed medically, with antibiotics if necessary, enteral or parenteral feeding and frequent control by imaging procedures. In 8 patients, the left drain was maintained in situ up to 43 days after surgery. In 4 patients, the drain had been removed between the 5th and 8th days after surgery after a normal radiologic study, but had to be inserted under radiological control 2-3 weeks after the gastric bypass. Daily output increased significantly the second week after surgery, and the leak closed at a mean of 30 days after surgery. One patient of the 12 (8%) died 32 days after surgery from septic shock, without any abdominal collection secondary to the leak. CONCLUSION: The occurrence of an anastomotic leak is nearly 2% after gastric bypass. The majority of them can be managed medically, without the need for a reoperation, due to the fact that there is no acid production in the small gastric pouch and there is no intestinal reflux due to the long Roux loop. PMID- 16259882 TI - Management of anastomotic leaks. PMID- 16259883 TI - Early improvement of glucose tolerance after ileal transposition in a non-obese type 2 diabetes rat model. AB - BACKGROUND: Surgical operations which shorten the intestinal tract between the stomach and the terminal ileum result in an early improvement in type 2 diabetes, and one possible explanation is the arrival of undigested food in the terminal ileum. This study was designed to evaluate the role of the distal ileum in the improvement of glucose control in type 2 diabetic patients who underwent bariatric surgery. METHODS: An ileal transposition (IT) to the jejunum was performed in lean diabetic Goto-Kakizaki (GK) rats. The IT was compared to sham operated diabetic rats and a control group of diabetic rats. Non-diabetic controls were age-matched Sprague-Dawley (SD) rats, which underwent IT and no operation. Food intake and body weight were measured. An oral glucose tolerance test (OGTT) was performed 10 days before the operation and 10 days, 30 days and 45 days after the surgery. GLP-1 and insulin were measured during the OGTT 45 days after surgery. An insulin tolerance test (ITT) was performed 50 days after surgery. RESULTS: Glucose tolerance improved in the IT diabetic group compared with both the sham-operated animals and control diabetic group 30 days and 45 days after surgery (P=0.029 and P=0.023, respectively). Insulin sensitivity, as measured by an ITT, was not significantly different between diabetic groups and the normal groups respectively after surgery. No differences in basal glucose and glucose tolerance were noted between non-diabetic operated animals and control non-diabetic rats. No differences were recorded between the diabetic rat groups and the non-diabetic rats in terms of weight and food intake. GLP-1 levels were significantly higher in the IT diabetic group compared with the sham-operated rats (P=0.05). CONCLUSIONS: Ileal transposition is effective in inducing an improvement in glucose tolerance in lean diabetic rats without affecting weight and food intake. The possible mechanism underlying the early improvement of diabetes after bariatric surgery may be due to the action of the terminal ileum through an insulin-independent action. PMID- 16259884 TI - Higher content of trans fatty acids in abdominal visceral fat of morbidly obese individuals undergoing bariatric surgery compared to non-obese subjects. AB - BACKGROUND: The purpose of this study was to determine the total content of trans fatty acids (TFA) in subcutaneous, retroperitoneal and visceral fat of morbidly obese and non-obese patients submitted to bariatric surgery or plastic and abdominal surgery. METHODS: The adipose tissues were obtained by surgery; lipids were extracted, saponified and esterified. TFA were measured by FTIR-ATR spectroscopy. RESULTS: The TFA average in obese patients was 6.3% for retroperitoneal and 8.7% for visceral fat. For non-obese patients, the figures were 6.9% (subcutaneous) and 9.3% (visceral). There was no difference between the groups. However, the TFA depot in visceral fat was higher than other fatty tissues for morbidly obese (P<0.001) and non-obese (P<0.05) patients. CONCLUSIONS: Our values for TFA content in all adipose tissues analyzed are higher than reported in other countries (3-6%). We showed more TFA in visceral adipose tissue than in other abdominal fat (subcutaneous and retroperitoneal) stores. The visceral adipose tissue level is worrisome because the higher rate of lipolysis in this tissue appears to be an important indicator of metabolic alterations and the levels of TFA found in adipose tissue presumably reflect the higher dietary intake of TFA by Brazilians. PMID- 16259885 TI - Comparison of bariatric and non-bariatric elective operations in morbidly obese patients on the basis of wound infection. AB - BACKGROUND: Wound infection rates after various types of bariatric operations have been well described. The question of whether bariatric surgery increases wound infection rate compared with similar elective surgical procedures in obese patients has not been clearly answered. The purpose of this study was to investigate wound status of morbidly obese patients after elective general surgery. METHODS: A prospective evaluation was conducted of 141 morbidly obese patients undergoing bariatric (n=60) and non-bariatric elective general surgery operation of similar invasiveness (n=81) with the ASEPSIS wound surveillance method. RESULTS: Median age of patients undergoing non-bariatric elective surgery (51, 32-68) was significantly higher than patients exposed to bariatric surgery (39, 24-57). Patients undergoing bariatric surgery had higher BMIs (44.0, 35 52.5) compared to the others (38.4, 35-43). All patients enrolled in the study were followed for a 21-day period. At the 7th postoperative day, 9 patients in the bariatric surgery group developed infection (15%), whereas 13 patients (16%) in the non-bariatric surgery group suffered wound infection. Wound infection was still present in 2 patients (3.4%) in the bariatric surgery group and 3 patients (3.7%) in the non-bariatric surgery group at the 21st day. Risk factors for wound infection included history of coronary artery diseases, diabetes, chronic respiratory illness and malignant disease. CONCLUSION: Bariatric surgery does not cause an additional risk of postoperative wound infection in morbidly obese patients, compared to elective general surgical operations of the same invasiveness. PMID- 16259886 TI - Wound infections in the morbidly obese. PMID- 16259887 TI - Use of gentamicin in the treatment of access-port infections. AB - BACKGROUND: A therapy concept for access-port infections is presented. METHODS: Between January 2001 and May 2005, 556 adjustable gastric bands were placed laparoscopically, and access-port infection data were analyzed. 6 early infections and 1 late infection occurred. 2 early infections were treated successfully with placement of a PMMA-chain at the port-site--without port removal. 2 other early infections were treated successfully with port removal and later reconnection; however, infection recurred at the access-port soon after reconnection, so a PMMA-chain was positioned around the port. The last 2 early infections were treated successfully by port removal and later connection of a new access-port surrounded by a PMMA-chain. The late access-port infection appeared to be caused by gastric erosion. RESULTS: Complete healing was achieved in all cases of early infection, and follow-up revealed no complications with subsequent band adjustments. The gastric erosion required removal of the entire banding system. CONCLUSION: For early port infection, the placement of a PMMA chain around the subcutaneous port appears to be a safe and effective approach that is less invasive than the usual port removal under general anesthesia. Placing the PMMA-chain is a rapid and simple procedure that allows retention of the original access-port. Once local healing is complete, the port can then be accessed easily and safely for band inflation. PMID- 16259888 TI - The impact of preoperative weight loss in patients undergoing laparoscopic Roux en-Y gastric bypass. AB - BACKGROUND: Laparoscopic Roux-en-Y gastric bypass (LRYGBP) is a widely performed bariatric operation. Preoperative factors that predict successful outcomes are currently being studied. The goal of this study was to determine if preoperative weight loss was associated with positive outcomes in patients undergoing LRYGBP. METHODS: A retrospective analysis was performed of all patients undergoing LRYGBP at our institution between July 2002 (when a policy of preoperative weight loss was instituted) and August 2003. Outcome measures evaluated at 1 year postoperatively included percent excess weight loss (EWL) and correction of co morbidities. Statistical analysis was performed by multiple linear regression. P<0.05 was considered significant. RESULTS: The study included 90 subjects. Initial BMI ranged from 35.4 to 63.1 (mean 48.1). Preoperative weight loss ranged from 0 to 23.8% (mean 7.25). At 12 months, postoperative EWL ranged from 40.4% to 110.9 % (mean 74.4%). Preoperative loss of 1% of initial weight correlated with an increase of 1.8% of postoperative EWL at 1 year. In addition, initial BMI correlated negatively with EWL, so that an increase of 1 unit of BMI correlated with a decrease of 1.34% of EWL. Finally, preoperative weight loss of >5% correlated significantly with shorter operative times by 36 minutes. Preoperative weight loss did not correlate with postoperative complications or correction of co-morbidities. CONCLUSIONS: Preoperative weight loss resulted in higher postoperative weight loss at 1 year and in shorter operative times with LRYGBP. No differences in correction of co-morbidities or complication rates were found with preoperative weight loss in this study. Preoperative weight loss should be encouraged in patients undergoing bariatric surgery. PMID- 16259889 TI - Prevalence of subclinical hypothyroidism in a morbidly obese population and improvement after weight loss induced by Roux-en-Y gastric bypass. AB - BACKGROUND: There are many studies concerning thyroid function in obesity, and some of them describe higher TSH levels in obese subjects. Few studies evaluated long-term changes in thyroid function caused by weight loss after bariatric surgery. Our aims were to evaluate the prevalence of subclinical hypothyroidism (SH) in a morbidly obese population and to analyze the effect of weight loss induced by Roux-en-Y gastric bypass (RYGBP) on TSH and thyroid hormone (TH) levels. METHODS: TSH, free thyroxine (fT4) and total triiodothyronine (T3) levels were analyzed before and 12 months after RYGBP in patients with grade III or grade II obesity with co-morbidities. Subjects taking TH and/or with positive antithyroid antibodies and/or with overt hypothyroidism were excluded. RESULTS: 72 subjects (62F/10M), with mean age 39.6+/-9.8 years and mean BMI 53.0+/-10.4 kg/m2 were studied. The prevalence of SH before RYGBP was 25% (n=18). There was a significant post-surgical decrease in BMI in the whole population, as well as in SH patients. In the SH group and normal TSH group, there was a decrease in TSH and T3, but not in fT4. TSH was not correlated with initial BMI or percent change in BMI. TSH concentrations reached normal values in all SH patients after RYGBP. CONCLUSION: Our data confirm that severe obesity is associated with increased TSH. The decrease in TSH was independent of BMI, but occurred in all SH patients. A putative effect of weight reduction on the improvement of SH in all patients may be an additional benefit of bariatric surgery. PMID- 16259890 TI - Laparoscopic adjustable gastric banding versus open vertical banded gastroplasty: a prospective randomized trial. AB - BACKGROUND: Laparoscopic adjustable gastric banding (LAGB) and open vertical banded gastroplasty (VBG) are treatment modalities for morbid obesity. However, few prospective randomized clinical trials (RCT) have been performed to compare both operations. METHODS: 100 patients (50 per group) were included in the study. Postoperative outcomes included hospital length of stay (LOS), complications, percent excess weight loss (%EWL), BMI and reduction in total comorbidities. Follow-up in all patients was 2 years. RESULTS: LOS was significantly shorter in the LAGB group. 3 LAGB were converted to open (1 to gastric bypass). Directly after VBG, 3 patients needed relaparotomies due to leakage, of which one (2%) died. After 2 years, 100% follow-up was achieved. BMI and %EWL were significantly decreased in both groups but significantly more in the VBG group compared to the LAGB group (31.0 kg/m2 and 70.1% vs 34.6 and 54.9% respectively). Co-morbidities significantly decreased in both groups in time. 2 years after LAGB, 20 patients needed reoperation for pouch dilation/slippage (n=12), band leakage (n=2), band erosion (n=2) and access-port problems (n=4). In the VBG group, 18 patients needed revisional surgery due to staple-line disruption (n=15), narrow outlet (n=2) or insufficient weight loss (n=1). Furthermore, 8 VBG patients developed an incisional hernia. CONCLUSION: This RCT demonstrates that, despite the initial better weight loss in the VBG group, based on complication rates and clinical outcome, LAGB is preferred. It had a shorter LOS and less postoperative morbidity. PMID- 16259891 TI - Laparoscopic vertical banded gastroplasty: 5-year results. AB - BACKGROUND: Vertical banded gastroplasty (VBG) has been a popular bariatric operation for the past 2 decades, and this operation has evolved into a laparoscopic procedure. However, reports of laparoscopic VBG (LVBG) from large series with longer results are limited. METHODS: From October 1998 to May 2002, 612 consecutive patients underwent LVBG. Mean age was 30.1 years and mean BMI 43.0 kg/m2. Laparoscopic Mason gastroplasty was performed. The change of BMI, obesity-related co-morbidities, and GI quality-of-life index (GIQLI) were studied. RESULTS: The major and minor complication-rate was 1.14% and 4.58% respectively. The mortality-rate was 0.16%. Mean BMI fell from 43.1 kg/m2 to 31.2, 31.3, 31.4, 32.2, and 32.8 kg/m2 at 1, 2, 3, 4, and 5 years respectively, with 93% follow-up. Revision rate was 9.2%. GIQLI decreased from 113.0 to 106.6, 110.9, 111.9, 112.1, and 106.4 at 1, 2, 3, 4, and 5 years. CONCLUSION: LVBG is safe and effective in weight reduction. The GIQLI failed to improve postoperatively even with good resolution of obesity-related co-morbidities. In carefully selected patients with diligent postoperative follow-up, LVBG is a bariatric surgery option. PMID- 16259892 TI - Continued excellent results with the mini-gastric bypass: six-year study in 2,410 patients. AB - BACKGROUND: There is a growing body of evidence showing that the Mini-Gastric Bypass (MGB) is a safe and effective alternative to other bariatric surgical operations. This study reports on the results of a consecutive cohort of patients undergoing the MGB. METHODS: A prospective database was used to continuously assess the results in 2,410 MGB patients treated from September 1997 to February 2004. RESULTS: The average operative time was 37.5 minutes, and the median length of stay was 1 day. The 30-day mortality and complication rates were 0.08% and 5.9% respectively. The leak rate was 1.08%. Average weight loss at 1 year was 59 kg (80% of excess body weight). The most frequent long-term complications were dyspepsia and ulcers (5.6%) and iron deficiency anemia (4.9%.) Excessive weight loss with malnutrition occurred in 1.1%. Weight loss was well maintained over 5 years, with <5% patients regaining more than 10 kg. CONCLUSIONS: Overall, the MGB is very safe initially and in the long-term. It has reliable weight loss and complications similar to other forms of gastric bypass. PMID- 16259893 TI - The comparison of scoring scales for liver biopsy assessment in morbidly obese patients undergoing bariatric surgery. AB - BACKGROUND: Many scoring systems have been applied for the grading and staging of non-alcoholic fatty liver disease (NAFLD). There is no consensus according to semiquantitative scales for the assessment of steatosis, inflammatory grading, and fibrosis staging in NAFLD. METHODS: We analysed 24 consecutive patients who underwent bariatric surgery. The grading for steatosis was estimated according to the systems proposed by Brunt and by Dixon. Brunt's scale and Scheuer's scale modified by Gabriel were used for inflammatory activity and fibrosis staging. Additionally, types of NAFLD disease were diagnosed according to Matteoni's classification. RESULTS: Steatosis was observed in 88% and steatohepatitis in 54% of patients. We observed portal, periportal and pericellular fibrosis. Neither bridging fibrosis nor cirrhosis were found. Extent of steatosis estimated according to Dixon and Brunt's scales was positively associated with appearance of steatohepatitis. The comparison of Dixon's and Brunt's scales according to grade of steatosis demonstrated a statistically significant difference. Inflammatory activity grades and fibrosis stages assessed according to Scheuer and Brunt scales differ significantly. Inflammatory activity evaluated with the Brunt scale was associated with the extent of steatosis and occurrence of steatohepatitis. CONCLUSIONS: Non-advanced forms of liver fibrosis do not appear to be dependent on steatosis and inflammatory grade in NAFLD. It is necessary to find the precise estimation of extent of steatosis especially occupying less than 1/3 or 1/4 of the lobule area. Brunt's scale seems to be more useful for the estimation of liver biopsy in NAFLD. It is essential to create a consensus for evaluation of steatosis and necroinflammatory grading and fibrosis staging in NAFLD. PMID- 16259894 TI - Non-alcoholic fatty liver disease: scoring systems need standardization, but are we ready? PMID- 16259895 TI - Efficacy of venous thromboembolism prophylaxis in morbidly obese patients undergoing gastric bypass surgery. AB - BACKGROUND: Patients undergoing gastric bypass are at risk of developing venous thromboembolism (VTE) due to multiple risk factors including obesity and abdominal surgery. The purpose of this study is to evaluate the effectiveness of inpatient VTE prophylaxis in morbidly obese patients undergoing gastric bypass and the incidence of symptomatic VTE following discharge. METHODS: A retrospective chart review of patients undergoing gastric bypass from August 2000 to August 2001 was performed. Inpatient charts from medical records and physician outpatient office charts were reviewed. Evaluation consisted of: VTE prophylaxis utilized, acquired risk factors for VTE, BMI, development of deep venous thrombosis (DVT) or pulmonary embolism (PE) during hospitalization, outpatient office visits following discharge, and VTE after discharge. RESULTS: 107 patient charts were reviewed. There were no incidences of VTE documented during hospitalization, and only one patient developed a symptomatic DVT after discharge. During hospitalization, all patients received DVT prophylaxis consisting of medical management, external compression devices, and ambulation orders. At the time of surgery, patients had a mean age of 40 years (23-69 years) and a BMI of 51.3 kg/m2 (37-82). Surgery lasted a mean of 108.9 minutes (65-305), patients were hospitalized for a mean of 4.3 days (3-7), and had a mean of 3.4 risk factors (2-7 risk factors) for the development of VTE. After hospital discharge, 101 patients were followed for the development of VTE. CONCLUSION: Combination of medical management, early ambulation, and external compression devices adequately prevented the development of VTE in patients after gastric bypass. PMID- 16259896 TI - Gastric electrical stimulation (GES) with parameters for morbid obesity elevates lower esophageal sphincter (LES) pressure in conscious dogs. AB - BACKGROUND: Treatment of obese patients with the implantable gastric stimulator (IGS) was reported to improve reflux symptoms, independent of weight loss. We evaluated the effect of gastric electrical stimulation on LES pressure in conscious dogs. METHODS: 8 dogs were studied. GES with three sets of parameters was randomly applied via a pair of electrodes implanted in the fundus on separate days. Manometry was performed with a Dent-Sleeve catheter passed through an esophageal canula. The involvement of the cholinergic pathway was also tested. RESULTS: 1) Stimulation with IGS parameters (40 Hz, 0.3 ms, 6 mA, 2 seconds on and 3 seconds off) induced a significant increase in LES pressure (29.9+/-4.8 mmHg), and remained significantly higher during the post-stimulation period (32.6+/-9.6 mmHg) compared to baseline (24.5+/-3.8 mmHg), P<0.01.2) Long pulse stimulation (10 cpm, 300 ms, 8 mA) tended to increase LES pressure from 29.6+/ 4.4 mmHg of baseline to 31.8+/-4.9 mmHg with stimulation, to 32.6+/-4.5 mmHg after discontinuation (P=0.08); 3) Modified IGS parameters (40 Hz, 2 ms, 6 mA, 2 seconds on and 3 seconds off) did not induce a significant change in LES pressure during and after stimulation. 4) Effect of stimulation with IGS parameters on LES pressure was blocked by intravenous atropine. CONCLUSION: GES with IGS parameters significantly increases LES pressure in conscious dogs. This effect is mediated by the cholinergic pathway. These results suggest that GES may be able to benefit GERD patients and obese patients with GERD. PMID- 16259897 TI - Laparoscopic colorectal surgery in obese patients. AB - BACKGROUND: The aim of the study was to the evaluate results of laparoscopic colorectal surgery in obese patients. METHODS: All patients who underwent elective laparoscopic colorectal surgery from January 1993 to December 2003 were included in the study. BMI>30 was used as an objective obesity criterion. The evaluated parameters included BMI, age, sex, diagnosis and associated diseases, American Society of Anesthesiologists classification score (ASA), type and duration of procedure, peroperative and postoperative complications, postoperative course, reoperation, length of hospitalization, morbidity and early mortality. RESULTS: 435 patients were evaluated. There were 80 patients (18%) in the obese group, and 355 patients (82%) were non-obese. The samples were comparable in terms of age, gender, ASA, diagnosis and procedure. Peroperative complications occurred more frequently in the obese group of patients (4% vs 2.5%, P>0.05) and the operating time was longer as well (151 min vs 141 min, P>0.05), both statistically not significant. There was no difference in postoperative course in both groups with regard to intravenous administration of analgesics (2 days), start of solid diet (day 3) and first bowel movement (day 4). Morbidity was higher in the obese group of patients (33% vs 24%, P>0.05), and reoperations were also more frequent here (13% vs 7%, P>0.05), which was reflected in prolonged hospital stay (14 days vs 12 days, P>0.05). On the other hand, early mortality was surprisingly lower in the obese group of patients (2.5% vs 6%, P>0.05). However, none of these differences achieved statistical significance on the set significance level of P=0.05. CONCLUSION: With sufficient experience, laparoscopic colorectal surgery in obese patients is feasible and safe. It is associated with no increased risk of complications and preserves all benefits of the mini-invasive approach. PMID- 16259898 TI - Laparoscopic Roux-en-Y gastric bypass with subtotal gastrectomy. AB - Laparoscopic gastric bypass is a common procedure for morbid obesity. After gastric bypass, the distal stomach is unavailable for surveillance. When a suspicious distal gastric lesion is present preoperatively, a distal subtotal gastrectomy may be needed. Herein we describe such a case performed laparoscopically. Laparoscopic gastric bypass with subtotal gastrectomy for morbid obesity should be considered for patients with suspicious distal gastric lesions. PMID- 16259900 TI - Intractable abdominal pain following laparoscopic adjustable gastric banding. AB - A 42-year-old woman presented with intractable crampy abdominal pain continuing for 1 year. The pain worsened after eating and improved when walking. She had undergone laparoscopic adjustable gastric banding 4 years previously, with re banding 18 months later because of anterior band slippage. The patient underwent numerous examinations, all with normal findings. A scintigraphy finally showed some tracer enrichment in the terminal ileum, which led to the assumption that a Meckel's diverticulum was causing her discomfort. Subsequent diagnostic laparoscopy showed no Meckel's diverticulum but instead displaced tubing, which was wrapped around the mesenteric root. The mesenteric root showed scarred alterations from chronic strangulation. After replacing the tubing from the band the abdominal pain immediately vanished. In retrospect, the contrast study of the gastric band shows unnatural traction of the tubing towards the lower abdomen, allowing suspicion of the intraoperative findings. PMID- 16259899 TI - Closure of the abdominal cavity after severe peritonitis in bariatric surgery utilizing a mesh and plastic device. AB - The major cause of peritonitis in bariatric surgery is leakage of GI contents, which can have a catastrophic outcome for the bariatric patient. To resolve this serious problem, the surgeon must act quickly. This paper describes a 27-year-old female after gastric bypass with disruption of the gastroenterostomy and severe contamination and peritonitis. Closure of the anastomotic leak, drainage, and gastrostomy in the bypassed stomach were performed, but the abdomen could not be closed, due to dilated bowel and the intra-abdominal edema with the sepsis. Temporary laparostomy closure was performed; a plastic sheet with an overlying mesh was sutured to the fascial margins. Planned multiple reoperations permitted removal of necrotic and infected debris, with progressive approximation and ultimate closure of the fascia. This treatment resulted in a successful outcome for the patient. PMID- 16259901 TI - Laparoscopic gastric banding for morbid obesity in a patient with situs inversus totalis. AB - We discuss the operative challenges posed by the advanced laparoscopic approach for a patient with situs inversus totalis. The patient was a morbidly obese woman with multiple co-morbidities related to her weight. The modifications in the surgical technique include the insertion of trocars according to the mirror image anatomy of the intra-abdominal organs under laparoscopic visualization. We suggest preoperative abdominal ultrasound in order to diagnose both gallbladder stones and also the reverse location of intra-abdominal organs that is rarely seen. A laparoscopic gastric banding, not a contradiction for situs inversus totalis, was performed successfully. PMID- 16259903 TI - How to improve office collection in a bariatric practice. AB - Office collections have always been under-valued in bariatric surgery because the money received is less than monies received from surgery. Co-payment must be obtained prior to the patient's evaluation, which will bring office collection to 99%. Such collections will cover costs and office charges. Virtually any consultation regarding bariatric surgery is high complexity, and should be charged as such. Morbidly obese patients usually have multiple disorders, each of which must be evaluated and taken into consideration. High cost is involved in operating a bariatric clinic; therefore, it is important to perform an accurate and detailed office analysis and collections for the economics of the practice. These profits will allow for needed personnel to be hired, covering the costs, and avoiding loss of income and time due to unwarranted cancellations. PMID- 16259902 TI - Rhabdomyolysis after biliopancreatic diversion with duodenal switch. AB - Rhabdomyolysis is an uncommon event in bariatric surgery. It can be caused by ischemia, crush injury, alcohol ingestion and drug intake, and as a consequence renal failure can develop. A few reports indicate that patients undergoing bariatric surgical intervention are at risk for rhabdomyolysis. A super-obese male (BMI 52 kg/m2) is reported, who underwent laparoscopic biliopancreatic diversion with duodenal switch (BPD/DS). Operative time was 265 minutes, and the BPD/DS operation was uneventful. Post-operatively, the patient complained of pain in both hips and the left shoulder, and suffered oliguria. He was treated with fluids (isotonic saline), bicarbonate, and mannitol. Despite this, he developed renal failure, which subsequently required hemodialysis. The patient died from arrhythmia and cardiac arrest on the 8th postoperative day. Obese patients undergoing bariatric surgery are at risk of rhabdomyolysis. Prolonged compression of the muscles during the surgical intervention, in long laparoscopic procedures, predisposes to this complication. PMID- 16259904 TI - William the Conqueror: war over weight. PMID- 16259905 TI - Prophylactic vena caval filters. PMID- 16259907 TI - Critical analysis of the staged laparoscopic Roux-en-Y: a two-stage operation to diminish the size of the liver in super-obese patients. PMID- 16259910 TI - [The combat casualty cure strategy under the new revolution in military affairs]. PMID- 16259909 TI - [Liver transplantation is laboring in the progress in China]. PMID- 16259911 TI - [Changes in pulmonary gas exchange and intrapulmonary shunt during orthotopic liver transplantation with non-venovenous bypass]. AB - OBJECTIVE: To study the changes in pulmonary gas exchange and intrapulmonary shunt during orthotopic liver transplantation (OLT) with non-venovenous bypass. METHODS: Nineteen American Society of Anesthesiologists (ASA) III-IV patients (male 17, female 2) with terminal liver diseases were enrolled for study. Their age ranged from 25-67 years. Anesthesia was induced with midazolam 0.05 mg/kg, propofol 0.5-1.0 mg/kg, fentanyl 4 microg/kg, with vecuronium 0.1 mg/kg, and it was maintained with isoflurane inhalation, fentanyl and vecuronium. All patients were mechanically ventilated with 100% O(2) during operation. After induction of anesthesia, Swan-Ganz catheter was inserted via right internal jugular vein. Cardiac output (CO), mixed venous oxygen saturation and core venous temperature were continuously monitored with continuous cardiac output monitor, and electrocardiogram (ECG), central venous pressure (CVP), pulmonary arterial wedge pressure (PAWP), pulse oxygen saturation (SpO(2)) and end-tidal carbon dioxide tension (P(ET)CO(2)) were also continuously monitored during operation. Radial artery was cannulated for continuous direct blood pressure monitoring. Arterial and mixed venous blood samples were taken after induction of anaesthesia, and partial pressure of oxygen (PaO(2)), partial pressure of carbon dioxide (PaCO(2)), and cardiac index(CI) were determined after induction of anaesthesia, 30 minutes before anhepatic stage, 30 minutes during anhepatic stage, 30 minutes during neohepatic stage and at the end of operation. Alveolar-arterial oxygen partial pressure difference (P(A-a)O(2)) and intrapulmonary shunt (Qs/Qt) were calculated according to the standard formula. RESULTS: After induction of anaesthesia, when the inspired oxygen flow (FiO(2)) was 1.00, PaO(2) was only (385.0+/-56.4) mm Hg (1 mm Hg=0.133 kPa), P(A-a)O(2) and Qs/Qt were all higher than normal values. There were no significant changes 30 minutes before anhepatic stage as compared with that after induction of anaesthesia. CO, CI and Qs/Qt were decreased significantly during anhepatic stage compared with that after induction of anaesthesia. PaO(2), PaCO(2), CO and CI were increased and P(A-a)O(2) decreased significantly, but there were no significant changes in Qs/Qt 30 minutes during neohepatic stage. CI and CO increased and Qs/Qt decreased significantly at the end of operation, but there were no significant difference in PaO(2), PaCO(2) and P(A-a)O(2). CONCLUSION: There are obvious changes in pulmonary gas exchange and intrapulmonary shunt during OLT with non-veno-venous bypass. PMID- 16259912 TI - [Model for predicting survival of patients with severe virus hepatitis and its clinical application]. AB - OBJECTIVE: To study prospectively the short-time survival of patients with severe virus hepatitis using model of severe liver diseases (MSLD) established by our previous study. METHODS: One hundred and three patients with severe hepatitis were included by cohort study. Of them, there were 85 patients with severe chronic hepatitis patients, 10 acute and 8 subacute severe hepatitis patients, respectively. The follow-up endpoint was 6 months. The cutoff score of the MSLD was determined by receiver operating characteristic cure (ROC) statistic analysis, and the survival of severe hepatitis patients in 2 weeks, 4 weeks, 3 months and 6 months were estimated by Kaplan-Meier statistic analysis. RESULTS: The cutoff MSLD score for predicting survival was 5. The survival curves of group A (total MSLD score< or =4) was significantly better than group B (total MSLD score> or =5, P<0.000). After treatment for 2 weeks, the survival rate in 2 weeks and 4 weeks was 37.9% and 3.5%, respectively, if MSLD score had no change or increased. The survival rate in 2 weeks, 4 weeks and 3 months was respectively 61.5%, 15.4%, 5.8% if the MSLD score decreased 1. Then, the survival in 2 weeks, 4 weeks, 3 months and 6 months was respectively 95.0%, 90.0%, 63.9% and 52.4% if MSLD score decreased 2 or more. CONCLUSION: It is suggested that MSLD may be valuable in predicting 6-month survival of severe virus hepatitis patients. It may be used to determine the efficacy of medical treatment and to guide clinical decision. PMID- 16259913 TI - [Analysis of peri-operative management in 1,510 patients with orthotopic liver transplantation]. AB - OBJECTIVE: To study the clinical factors and treatment that related to early prognosis of patients after orthotopic liver transplantation (OLT), in order to improve peri-operative care and raise the survival rate in patients undergoing this operation. METHODS: Clinical records of 1,510 patients who had undergone OLT in our center were retrospectively analyzed. Twenty variables in pre-operative, intra-operative or post-operative periods were included, and several risk factors which affected the early prognosis after OLT were screened. RESULTS: With univariate analysis, the following variables showed significant difference between the survival group and the dead group: acute pathology and chronic health evaluation III (APACHE III) score, volume of transfusion and oxygen delivery (DO(2)) of tissue during operation, incidence of infection, and demand of blood purification. CONCLUSION: During peri-operative periods of OLT, combined therapy should be used, and attention should be paid to the risk factors. PMID- 16259914 TI - [Changes in hemodynamics and nitric oxide/endothelin-1 during liver transplantation in patients with cirrhosis]. AB - OBJECTIVE: To study the systemic and pulmonary hemodynamic changes of patients with cirrhosis during liver transplantation and evaluate the role of nitric oxide (NO) and endothelin-1(ET-1). METHODS: Twenty-four patients with cirrhosis at terminal stage underwent modifying piggy-back liver transplantation. Hemodynamic parameters including cardiac index (CI), arterial blood pressure (ABP) and pulmonary arterial pressure (PAP) were monitored continuously. NO and ET-1 levels were measured by radioimmunoassay. Blood samples were obtained from superior vena cava at induction of anesthesia (T1), 10 minutes before vascular cross clamping (T2), 30 minutes after vascular cross clamping (T3), 30 minutes after reperfusion of the new liver (T4), and at the end of surgery (T5). RESULTS: (1) Mean arterial blood pressure (MABP) lowered significantly in the early stage of anhepatic period and neohepatic period (P<0.05 or P<0.01). (2) Central venous pressure (CVP), mean pulmonary arterial pressure (MPAP) and pulmonary arterial wedge pressure (PAWP) lowered significantly during anhepatic period. They rose significantly after graft reperfusion, and remained at a high level with respect to the baseline level (P<0.05). (3) CI declined significantly during anhepatic period and increased 10 minutes after reperfusion of new liver. (4) Systemic vascular resistance index and pulmonary vascular resistance index increased during anhepatic period and were higher than the baseline level 15 minutes after reperfusion. SVRI was lower than baseline level 30 minutes after reperfusion. (5) Compared with the baseline level, NO decreased significantly after vascular cross clamping and elevated 30 minutes after reperfusion. ET levels were significant elevated 30 minutes after clamping and after reperfusion (P<0.05). CONCLUSION: Significant hemodynamic changes occur in patients with cirrhosis during liver transplantation, and pulmonary hypertension develops during neohepatic period. The role of elevated contents of NO and ET-1 after reperfusion needs further study. PMID- 16259915 TI - [Changes in oxygen metabolism in hepatitis B gravis in peri-operative stages of orthotopic liver transplantation]. AB - OBJECTIVE: To investigate the changes in oxygen metabolism in peri-operative stages in hepatitis B gravis and non-hepatitis B gravis undergoing orthotopic liver transplantation (OLT). METHODS: Twelve patients undergoing OLT for hepatitis B gravis (experimental group) and 10 patients without hepatitis B gravis (control group) were enrolled for study. Anesthesia was induced by propofol, fentanyl and vecuronium bromide, and maintained with isoflurane. Arterial catheter was inserted into the left radial artery. Swan-Ganz catheter was inserted through the right internal jugular vein. Arterial partial pressure of oxygen (PaO(2)), mixed venous partial pressure of oxygen (PvO(2)), arterial oxygen content (CaO(2)), mixed venous oxygen content (CvO(2)), arterial-venous oxygen content difference (Ca-vO(2)), oxygen delivery (DO(2)), index of oxygen delivery (DO(2)I), oxygen consumption (VO(2)), index of oxygen consumption (VO(2)I), index of oxygen extract (O(2)EI), oxygen extract rate (O(2)ER) were determined before skin incision (T1), 10 minutes before anhepatic phase (T2), 25 minutes after liver was removed (anhepatic phase, T3), 30 minutes in neohapitic phase (T4), and the end of operation (T5). RESULTS: (1) In the experimental group, PvO(2) increased, Ca-vO(2) decreased, DO(2) and VO(2) showed no change, and O(2)EI and O(2)ER decreased in T2. In preanhepatic period, PvO(2) increased, Ca-vO(2) decreased, DO(2) and VO(2) did not change. In anhepatic period, DO(2), DO(2)I, VO(2) and VO(2)I decreased obviously, DO(2) decreased by 43%, while VO(2) decreased by 21%, and O(2)ER increased obviously. In reperfusion period, PaO(2) and PvO(2) increased, CaO(2) and Ca-vO(2) decreased, DO(2) and DO(2)I increased, VO(2) and VO(2)I recovered to base level. After termination of operation, PvO(2), DO(2), DO(2)I were still high. (2) In the control group: PvO(2) increased, O(2)EI and O(2)ER decreased, but no significant changes were found in DO(2) and VO(2) in T2. DO(2), DO(2)I, VO(2), and VO(2)I all decreased in T3, while DO(2) reduced by 25% and VO(2) reduced by 12%. In T4, PvO(2), DO(2) and DO(2)I all increased, while Ca-vO(2), VO(2) and VO(2)I reached the pre-operative levels in T4. DO(2) and DO(2)I levels were higher than those of pre-operation in T5. CONCLUSION: The decrease in DO(2) is more obvious than VO(2) in anhepatic period during OLT in hepatitis B gravis patients. In neohepatic period, DO(2) increases while VO(2) returns to base level. PMID- 16259917 TI - [A pilot study of using pure albumin as a dialysate in the treatment of liver failure]. AB - OBJECTIVE: To observe the effect of a new extracorporeal hepatic support hemodialysis with albumin-based dialysate on patient with liver failure, and to compare the result with that of molecular absorbent recycling system (MARS). METHODS: Eighteen patients with liver failure were enrolled and treated intermittently with artificial liver on the basis of conventional regime. Among them, 12 of them were treated with single albumin dialysis (SAD) for 6 hours each period, and 6 with MARS for 6 hours. During each session of SAD, 4,000 ml albumin based dialysate (45 g/L) was circulated in the dialysate route at a flow rate of 10 L/h with the blood flow rate of 250 ml/min. Liver and renal function, prothrombin activity (PTA), serum ammonia (NH(3)), endotoxin were monitored before and after the treatment, and the serial bilirubin content was measured in the serum and the albumin dialysate of all patients respectively 1-hour, 3-hour, 6-hour after the treatment with SAD. RESULTS: After treatment with SAD, the clinical symptoms and signs were improved and the hemodynamics were stabilized, complications were ameliorated partially, in particular hepatic encephalopathy was improved, with low incidence of adverse reaction, and the effective rate and survival rate were high. The serum PTA was increased obviously (P<0.05), and the serum blood urea nitrogen (BUN), serum creatinine (SCr), total bilirubin (TB), total bile acid (TBA), NH(3), endotoxin were decreased significantly (all P<0.05), and elimination of bilirubin slowed down gradually. The results showed that SAD was as effective as MARS, with a lower cost, and simpler and more convenient than MARS. CONCLUSION: Single albumin dialysis may eliminate protein bound and water-soluble hepatic toxins, improve clinical symptoms and liver function in the treatment of liver failure, and worth further clinical study. PMID- 16259918 TI - [Protective effect of protease inhibitor on renal function of rat challenged by lipopolysaccharide]. AB - OBJECTIVE: To investigate the protective effects of protease inhibitor on lipopolysaccharide (LPS)-induced kidney injury in rats and its possible mechanism. METHODS: Thirty-two male Wistar rats were divided into four groups randomly: normal control (n=6); LPS group (n=10), receiving intravenous endotoxin (LPS, O55:B5, 5 mg/kg); low-dose ulinastatin (UT) intervention group (n=8), receiving intraperitoneal UT 50 000 U/kg and LPS 5 mg/kg as above; high-dose UT intervention group (n=8), UT 100,000 U/kg and LPS 5 mg/kg. The following examinations were carried out: blood gas analysis, kidney pathological changes, plasma endothelin-1, plasma lactic acid, N-acetylglucosaminidase (NAG) in urine, and plasma creatinine (Cr) level. RESULTS: Blood gas analysis showed that pH, partial pressure of oxygen in artery (PaO(2)) and base excess (BE) lowered significantly (all P<0.01) in LPS group compared with normal control group, and elevated in low-dose and large dose UT intervention groups compared with LPS group, the differences were statistically significant (P<0.01). The plasma endothelin-1 increased significantly (P<0.01) in LPS group compared with normal control group and intervention groups. Plasma lactate increased significantly in LPS group compared with normal control group (P<0.001), decreased significantly in intervention groups compared with LPS group (P<0.01). No significant difference was found between two intervention groups (P>0.05). Plasma Cr and urine NAG level increased in LPS groups, and the difference was significant compared with normal control groups and intervention groups (P<0.01). Pathohistologic examination revealed normal glomeruli but vacuolar degeneration of tubular epithelial cells, and part of them disrupted and desquamated, and also tubular dilatation. Only mild pathological changes were seen in the intervention groups. There was no obvious difference in morphology between two intervention groups. CONCLUSION: The protease inhibitor, UT, may alleviate LPS-induced inflammatory reaction and damage to kidney in rats. PMID- 16259919 TI - [Effect of p21 on the changes in renal tubular epithelial cells after ischemia/reperfusion injury of kidney]. AB - OBJECTIVE: To investigate the contribution of p21 gene in renal tubular epithelial cells in p21 (+/+) and p21 (-/-) mice of young and old ages at different times after kidney ischemia/reperfusion injury (IRI). METHODS: In p21 (+/+) and p21 (-/-) male mice at the ages of 2 and 12 months the kidneys were made ischemic by clamping the left renal artery for 45 minutes followed by declamping. On 0, 1, 3 and 7 days, 1, 3 and 6 months after reflow, renal tissue was processed for pathological study, determination of proliferating cell nuclear antigen (PCNA), apoptosis and senescence-associated beta-galactosidase (SA-beta gal) analysis, using hematoxylin and eosin staining, immunohistochemistry, terminal deoxynucleotidyl transferase-mediated dUTP-biotin in situ nick-end labeling (TUNEL), and histochemical staining, respectively. RESULTS: Renal tubule necrosis and cell apoptosis were more severe in p21 (-/-) mice and old mice as compared with p21 (+/+) mice and young mice (both P<0.05), respectively. In young p21 (+/+) mice, occasionally faint staining for SA-beta-gal activity began to appear after 1 month, and significantly increased 3 and 6 months after IRI (P<0.05), but there was no positive staining for SA-beta-gal in the contralateral kidney or both kidneys in p21 (-/-) mice at any time. Another manner of the expression of SA-beta-gal was detected in aged p21 (+/+) mice, as both kidneys showed intensely positive staining for SA-beta-gal at 0 day after IRI, it then subsided notably on 1 day in the IRI kidney (P<0.05), but increased again at 3 months, though still less intense than the contralateral kidney, albeit more intense than the young mice at the same time (P<0.05). Three months after IRI, in both the IRI kidney and the contralateral kidney, positive staining for SA-beta gal almost reached the same level. On the contrary, only occasional faint staining for SA-beta-gal activity was observed in aged p21 (-/-) mice at any time. No significant difference in positive staining of nuclear PCNA was found between in young and aged p21 (+/+) mice (P>0.05), although the numbers of positively stained nuclear PCNA were more in number in young mice than in aged mice. But in p21 (-/-) mice, significantly more cells were positively stained for PCNA, especially in young mice and in IRI kidneys (P<0.05). Correlation analysis between senescent and apoptotic cells in aged mice made at 1 day after IRI showed striking negative correlation between both of them [p21 (+/+) mice: r=-0.82; P<0.001; p21 (-/-) mice: r=-0.76, P<0.001]. CONCLUSION: IRI can promote the senescence process of normal tubular cells, and can accelerate death (necrosis and apoptosis) process of senescent tubular cells. p21 gene may play an important role in the senescence changes in tubular epithelial cells after kidney ischemia/reperfusion injury. PMID- 16259921 TI - [Expression of peroxisome proliferators-activated receptor gamma and alpha-smooth muscle actin in unilateral ureteral obstruction in rats]. AB - OBJECTIVE: To evaluate the expression of peroxisome proliferators-activated receptor gamma (PPAR gamma) and alpha-smooth muscle actin (alpha-SMA) in unilateral ureteral obstruction (UUO) in rats and the relationship between them and the extent of tubulointerstitial injury. METHODS: Male SD rats were randomly subjected to either left ureteral ligation (n=30) or sham operation (n=6), and they were sacrificed on 3, 7, 14, 21 or 28 days after UUO. Sections of renal tissue were stained with hematoxylin and eosin (HE), Masson, or periodic acid silver methenamine (PASM). The extent of tubulointerstitial injury was determined by Banff classification. Immunohistochemical staining was performed to investigate the expression of PPAR gamma and alpha-SMA in the renal tissue. RESULTS: Swelling of tubular epithelia cells, infiltration of inflammatory cells and proliferation of fibroblasts were not so obvious on days 3 and 7 after UUO. A diffuse inflammatory cells infiltration, and massive fibrous hyperplasia were detectable on days 14 and 21 after UUO. Most tubules showed serious damage and replaced by proliferative fibrous tissue on day 28. The expression of PPAR gamma was almost undetectable in sham operation group. However, it was upregulated on day 3 and peaking on day 14, and then it was slightly decreased on days 21 and 28. The expression of alpha-SMA was only found in vascular smooth muscle cells in sham operation group. It was upregulated mainly in some interstitial cells on day 3 and increased with the progression of tubulointerstitial fibrosis. CONCLUSION: PPAR gamma expression increases in rat renal tissue after UUO, but does not always correlate with the extent of the alpha-SMA expression and tubulointerstitial injury. These data suggest that increase in PPAR gamma in renal tissue may play an important role in response to inflammation and fibrosis. PMID- 16259923 TI - [Comparison of clinical characteristics between hospital-acquired and community acquired acute renal failure]. AB - OBJECTIVE: To investigate the differences between hospital-acquired acute renal failure (HA-ARF) and community-acquired acute renal failure (CA-ARF) in epidemiology, etiology and prognosis. METHODS: The diagnosis of ARF of patients diagnosed with ARF from ICD-9 codes, who were discharged from Peking University Third Hospital from January 1994 to December 2003, was reconfirmed and categorized by two nephrologists independently. The indexes of epidemiology, etiology and prognosis were studied. Single-variable analysis and multivariate logistic regression analysis were performed to investigate the correlation between clinical features and prognosis respectively. RESULTS: Two hundred and five cases were collected and all were reconfirmed. CA-ARF had a predominance of 59.5%. HA-ARF, however, increased by 1.06 times in last 5 years (P<0.05); 59.0% of HA-ARF was diagnosed in department of surgery while 70.5% of CA-ARF was in medical department (both P<0.05); 36.1% HA-ARF patients had two or more pathogenic causes, while 91.2% CA-ARF only had one cause (P<0.05); 49.4% HA-ARF developed after operation; 26.5% HA-ARF and 18.8% CA-ARF were drug-related (P>0.05); 24.1% HA-ARF and 12.3% CA-ARF were infection-related (P=0.028). Mortality and recovery rates were 62.7% and 20.6%, respectively, in HA-ARF while 23.0% and 67.2% in CA-ARF respectively (both P<0.01). The percentage of oliguria, multiple organ failure (MOF), systemic inflammatory response syndrome (SIRS), and use of mechanical ventilation were significantly higher in HA-ARF than in CA-ARF (all P<0.01). Acute tubule necrosis-injury severity score (ATN-ISS), acute pathological and chronic health evaluation (APACHE II) score were 0.54+/-0.24 and 19.6+/-4.9 in HA-ARF, while they were 0.27+/-0.18 and 15.7+/-5.6 in CA-ARF (both P<0.01). Multiple regression analysis identified that both MOF and SIRS were common independent risk factors for HA-ARF and CA-ARF, and oliguria and advanced age were respective independent risk factor for HA-ARF and CA-ARF. CONCLUSION: CA ARF prevails in hospitalized Chinese patients during the last 10 years, but HA ARF is increasing in incidence significantly during the last 5 years. The etiology is mostly simple and the prognosis is relatively good in CA-ARF, while the pathogenic cause is mostly complicated and the outcome is much poorer in HA ARF. PMID- 16259924 TI - [Replication of a model of injury to human renal proximal tubular cells induced by hypoxia/reoxygenation]. AB - OBJECTIVE: To replicate a new model of injury to human renal proximal tubular cells (HK-2) induced by hypoxia/reoxygenation. METHODS: Human renal proximal tubular cell line HK-2 cell was used as the target cell. Tubular cells were divided into six groups: 4 hours of hypoxia, 12 hours of hypoxia, 24 hours of hypoxia, and 24 hours of hypoxia followed by reoxygenation 4, 12 or 24 hours later groups. Each group was accompanied by a control group. Hypoxic culture conditions were produced by covering the culture with liquid paraffin. Trypan blue exclusion was used for cell count and cell viability. The activity of lactate dehydrogenase (LDH) in the culture medium was determined by biochemical method. RESULTS: After being challenged by hypoxia followed by reoxygenation, trypan blue exclusion rate was greater, cell count and cell viability were lower, and the activity of LDH was increased. It indicated that the destruction of integrity of cellular membrane was induced by ischemia/reperfusion injury, and the tubular cells may be injured irreversibly. CONCLUSION: A simple model of hypoxic injury of renal tubular cells is replicated by covering the culture cells with liquid paraffin. PMID- 16259926 TI - [Role of interleukin-1 receptor antagonist in protecting kidney from injury induced by asphyxia in neonatal rats]. AB - OBJECTIVE: To observe the role of recombinant human interleukin-1 receptor antagonist (rhIL-1ra) in protecting kidney from injury induced by asphyxia in neonatal rats. METHODS: Neonatal rats were used as experimental animals. The changes in intrarenal inflammatory response and renal injury were examined in the control group (n=13), and 2, 24 and 48 hours after asphyxia followed by normal saline treatment in those treated with rhIL-1ra. RESULTS: In normal saline group, the white blood cell count, the blood interleukin-1 (IL-1), IL-8, IL-6, nitric oxide (NO), endothelin (ET-1) levels, and the renal coefficient (LRC), the scores of injured tubules of the left kidney were significantly increased at 2 hours (n=10), 24 hours (n=11), and 48 hours (n=10, P<0.05 or P<0.01). Compared with the normal saline group, the levels of the above parameters, except IL-6, were significantly decreased in rhIL-1ra treatment group at the same time points (P<0.05 or P<0.01). Serum IL-6 at 24 hours and 48 hours was also decreased in rhIL-1ra treatment group significantly (P<0.05 or P<0.01). CONCLUSION: The results suggest that rhIL-1ra may protect renal injury after asphyxia via inhibiting intrarenal inflammatory response. PMID- 16259927 TI - [Changes in serum contents of inflammatory mediators after tetramethylene-disulfo tetramine poisoning and clinical study of a new treatment regime for the poisoning]. AB - OBJECTIVE: To study the changes in serum contents of beta-endorphin (beta-EP), endothelins (ET), nitric oxide (NO) and tumor necrosis factor (TNF) after acute tetramethylene-disulfo-tetramine (TDT) poisoning and therapeutic effect of a new treatment regime. METHODS: (1) Forty-eight patients with tetramethylene-disulfo tetramine poisoning (experiment group) were enrolled in this study. The serum levels of beta-EP, ET, NO and TNF were measured upon hospitalization and 1, 3, 5, 7, 9, 11, 13, 15, 17 and 19 days after poisoning, respectively, and compared with those of 30 healthy individuals (control group B). (2) They were treated with the improved regime and compared with patients treated with the conventional regime designated as control group A. RESULTS: (1) In 48 patients treated with improved regime, 45 were cured and 3 died. (2) The serum levels of beta-EP, ET, NO and TNF from 45 patients who were cured were significantly higher at hospitalization compared with those of healthy individuals, with the peak values appeared on day 1 after poisoning in the mild, moderate and severe groups. Beta-EP levels returned to normal range on days 9, 13 and 17 after poisoning respectively in the mild, moderate and severe groups. ET levels returned to normal range on days 7, 13 and 15 after poisoning respectively in the mild, moderate and severe groups. NO levels returned to normal range on days 7, 11 and 11 after poisoning respectively in the mild, moderate and severe groups. TNF levels returned to normal range on days 9, 11 and 17 after poisoning respectively in the mild, moderate and severe groups. (3) The serum levels of beta-EP, ET, NO and TNF in 3 non-survivors were very high at hospitalization and continued to increase in the course of treatment. (4) The cumulative doses of diazepam and Phenobarbital, and the eclampsia time were significantly less in the experiment group than those of control group A. CONCLUSION: (1) The serum levels of beta-EP, ET, NO and TNF are correlated with the severity of tetramethylene-disulfo-tetramine poisoning and general conditions of the patients. (2) When the serum levels of beta-EP, ET, NO and TNF decrease gradually in the course of treatment, prognosis is better. On the contrary, the prognosis is poor when their levels increase gradually. (3) Measures to decrease levels of beta-EP, ET, NO and TNF result in a better prognosis of patients with tetramethylene-disulfo-tetramine poisoning. (4) The improved regime can be considered a better therapeutic strategy in tetramethylene disulfo-tetramine poisoning. PMID- 16259936 TI - Isotopic labeling of proteins by utilizing photosynthetic bacteria. PMID- 16259937 TI - A zymogram method for detecting carrageenase activity. PMID- 16259938 TI - Multicommuted optosensor for the determination of pipemidic acid in biological fluids. PMID- 16259939 TI - Brushes with sage. AB - Wherein we learn that when working with plant terpene synthases, sometimes it is not the isotopes that get scrambled. PMID- 16259940 TI - An immunohistochemical study of the expression of heat-shock protein-25 and cell proliferation in the dental pulp and enamel organ during odontogenesis in rat molars. AB - OBJECTIVES: The aim of this study is to clarify the functional significance of heat-shock protein (HSP)-25 during tooth development. DESIGN: We compared the expression of HSP-25 in the dental epithelial and mesenchymal cells with their proliferative activity during odontogenesis in rat molars on postnatal days 1-100 by immunohistochemistry using anti-HSP-25 and anti-5-bromo-2'-deoxyuridine (BrdU) for cell proliferation assay. RESULTS: On day 1, BrdU-immunoreactive cells were densely located in the inner enamel epithelium in the cervical loop and intercusped areas and the dental pulp adjacent to them, whereas HSP-25 immunoractivity (IR) was restricted to the cusped area where odontoblasts and ameloblasts had already differentiated. Subsequently, BrdU-IR shifted in the apical direction to be localized around Hertwig's epithelial root sheath during days 5-30, never overlapping with concomitantly apically-shifted HSP-25-IR. On days 60-100, BrdU-immunoreactive cells were hardly recognizable in the dental pulp, where HSP-25-IR was exclusively localized in the odontoblast layer. Furthermore, the odontoblast- and ameloblast-lineage cells exhibited two steps in the expression of HSP-25 throughout the postnatal stages: first, dental epithelial and pulpal mesenchymal cells showed a weak IR for HSP-25 after the cessation of their proliferative activity, and subsequently odontoblasts and ameloblasts consistently expressed an intense HSP-25-IR. CONCLUSION: Odontoblast- and ameloblast-lineage cells acquire HSP-25-IR after they complete their cell division, suggesting that this protein acts as a switch between cell proliferation and differentiation during tooth development. The consistent expression of HSP-25-IR in the formative cells may be involved in the maintenance of their functional integrity. PMID- 16259941 TI - Gap junctional communication in the male reproductive system. AB - Male fertility is a highly controlled process that allows proliferation, meiosis and differentiation of male germ cells in the testis, final maturation in the epididymis and also requires functional male accessory glands: seminal vesicles, prostate and corpus cavernosum. In addition to classical endocrine and paracrine controls, mainly by gonadotropins LH and FSH and steroids, there is now strong evidence that all these processes are dependent upon the presence of homocellular or heterocellular junctions, including gap junctions and their specific connexins (Cxs), between the different cell types that structure the male reproductive tract. The present review is focused on the identification of Cxs, their distribution in the testis and in different structures of the male genital tract (epididymis, seminal vesicle, prostate, corpus cavernosum), their crucial role in the control of spermatogenesis and their implication in the function of the male accessory glands, including functional smooth muscle tone. Their potential dysfunctions in some testis (spermatogenic arrest, seminoma) and prostate (benign hyperplasia, adenocarcinoma) diseases and in the physiopathology of the human erectile function are also discussed. PMID- 16259942 TI - Habit reversal versus supportive psychotherapy in Tourette's disorder: a randomized controlled trial and predictors of treatment response. AB - Tourette syndrome (TS) is characterized by chronic motor and vocal tics. Habit reversal therapy (HR) is a behavioral treatment for tics which has received recent empirical support. The present study compared the efficacy of HRT in reducing tics, improving life-satisfaction and psychosocial functioning in comparison with supportive psychotherapy (SP) in outpatients with TS. In addition, we investigated whether impairments in response inhibition in patients with TS predict response to HR treatment which specifically aims to inhibit tics. Thirty adult outpatients with DSM-IV TS were randomized to 14 individual sessions of HR (n = 15); or SP (n = 15). HR but not SP reduced tic severity over the course of the treatment. Both groups improved in life-satisfaction and psychosocial functioning during active treatment. Reductions in tic severity (HR) and improvements in life-satisfaction and psychosocial functioning (HR and SP) remained stable at the 6-month follow-up. The extent of pre-treatment response inhibition impairment in the HR group predicted reductions in tic-severity from pre- to post-treatment. Our results suggest that HR has specific tic-reducing effects although SP is effective in improving life-satisfaction and psychosocial functioning. Assessments of response inhibition may be of value for predicting treatment response to HR. PMID- 16259943 TI - Sphingosine releases Ca2+ from intracellular stores via the ryanodine receptor in sea urchin egg homogenates. AB - Various reports have demonstrated that the sphingolipids sphingosine and sphingosine-1-phosphate are able to induce Ca2+ release from intracellular stores in a similar way to second messengers. Here, we have used the sea urchin egg homogenate, a model system for the study of intracellular Ca2+ release mechanisms, to investigate the effect of these sphingolipids. While ceramide and sphingosine-1-phosphate did not display the ability to release Ca2+, sphingosine stimulated transient Ca2+ release from thapsigargin-sensitive intracellular stores. This release was inhibited by ryanodine receptor blockers (high concentrations of ryanodine, Mg2+, and procaine) but not by pre-treatment of homogenates with cADPR, 8-bromo-cADPR or blockers of other intracellular Ca2+ channels. However, sphingosine rendered the ryanodine receptor refractory to cADPR. We propose that, in the sea urchin egg, sphingosine is able to activate the ryanodine receptor via a mechanism distinct from that used by cADPR. PMID- 16259944 TI - Downregulation of cyclin-dependent kinase inhibitor; p57(kip2), is involved in the cell cycle progression of vascular smooth muscle cells. AB - Immature vascular smooth muscle cells (VSMCs) proliferate responding to extrinsic mitogens and accumulate in neointima after arterial injuries. Cell proliferation is positively regulated by cyclin/cyclin-dependent kinase (CDK) complex and negatively controlled by CDK inhibitors; CKIs such as p27(kip1) and p57(kip2). In this study, embryonic rat thoracic aorta VSMCs; A10 were G0/G1 arrested by serum starvation, re-stimulated with serum, and harvested every four hours. Both CKIs co-expressed in quiescent VSMCs and rapidly diminished by stimulation. Protein level of p27(kip1) was regulated by both transcription and post-transcription, but that of p57(kip2) was mainly by post-transcription. Supplemental overexpression of p57(kip2) inhibited the activations of G1 cyclin/CDKs and subsequent hyperphosphorylations of all three retinoblastoma pocket proteins as well as G1/S transition of cell cycle. Our findings suggest that the downregulations of not only p27(kip1), but also p57(kip2) responding to mitogenic stimulation, play key roles in the cell cycle progression of VSMCs. PMID- 16259945 TI - A role for 3-O-sulfotransferase isoform-4 in assisting HSV-1 entry and spread. AB - Many heparan sulfate (HS) 3-O-sulfotransferase (3-OST) isoforms generate cellular receptors for herpes simplex virus type-1 (HSV-1) glycoprotein D (gD). Interestingly, the ability of 3-OST-4 to mediate HSV-1 entry and cell-to-cell fusion has not been determined, although it is predominantly expressed in the brain, a primary target of HSV-1 infections. We report that expression of 3-OST-4 can render Chinese hamster ovary K1 (CHO-K1) cells susceptible to entry of wild type and a mutant (Rid1) strain of HSV-1. Evidence for generation of gD receptors by 3-OST-4 was suggested by gD-mediated interference assay and the ability of 3 OST-4 expressing CHO-K1 cells to preferentially bind HSV-1 gD, which could be reversed by prior treatment of cells with HS lyases (heparinases-II/III). In addition, 3-OST-4 expressing CHO-K1 cells acquired the ability to fuse with cells expressing HSV-1 glycoproteins. Demonstrating specificity, the cell fusion was inhibited by soluble 3-O-sulfated forms of HS, but not unmodified HS. Taken together our results suggest a role of 3-OST-4 in HSV-1 pathogenesis. PMID- 16259946 TI - Efficient targeting of adenoviral vectors to integrin positive vascular cells utilizing a CAR-cyclic RGD linker protein. AB - Vascular smooth muscle (VSMC) and endothelial cells (EC) are particularly resistant to infection by type 5 adenovirus (Ad) vectors. To overcome this limitation and target Ad vectors to ubiquitously expressed alpha(V)beta(3/5) integrins, we have generated a linker protein consisting of the extracellular domain of the coxsackie adenovirus receptor (CAR) connected via avidin to a biotinylated cyclic (c) RGD peptide. After optimization of CAR to cRGD and to Ad coupling, infection of mouse heart endothelial cells (H5V) could be augmented significantly, as demonstrated by 600-fold increased transgene expression levels. In EOMAs, a hemangioendothelioma-derived cell line, the fraction of infected cells was enhanced 4- to 6-fold. Furthermore, the fraction of infected primary mouse VSMC was increased from virtually 0% to 25%. Finally, in human umbilical vein endothelial cells, the number of GFP positive cells was enhanced from 2% to 75%. In conclusion, CAR-cRGD is a versatile and highly efficient construct to target Ad vectors to both transformed and primary VSMC and EC. PMID- 16259947 TI - Co-culture of osteoblasts and chondrocytes modulates cellular differentiation in vitro. AB - Biological integration of cartilage grafts with subchondral bone remains a significant clinical challenge. We hypothesize that interaction between osteoblasts and chondrocytes is important in regenerating the osteochondral interface on tissue-engineered osteochondral grafts. We describe here a sequential co-culturing model which permits cell-cell contact and paracrine interaction between osteoblast and chondrocytes in 3-D culture. This model was used to determine the effects of co-culture on the phenotypic maintenance of osteoblasts and chondrocytes. It was found that while chondrocytes synthesized a type II collagen and glycosaminoglycan (GAG) matrix, GAG deposition was significantly lower in co-culture. Alkaline phosphatase activity was maintained in osteoblasts, but cell-mediated mineralization in co-culture was markedly lower compared to osteoblast controls. These results collectively suggest that interactions between osteoblasts and chondrocytes modulate cell phenotypes, and the importance of these interactions on osteochondral interface regeneration will be explored in future studies. PMID- 16259948 TI - ATP binding is required for physiological activation of retinal guanylate cyclase. AB - ATP bound to retinal guanylate cyclase (retGC)/membranes prior to the assay (pre binding effect) and during the assay (direct effect) further enhances retGC activity stimulated by GC-activating proteins (GCAPs). Here we investigate differences between these two effects. We found that the pre-binding effect, but not the direct effect, was absent in membranes pre-washed with Mg(2+)-free hypotonic buffers, that the pre-binding effect, but not the direct effect, was strictly limited to GCAP-stimulated retGC activity, and that these two effects were independent and additive rather than being synergistic. Pre-incubation with amiloride enhanced GCAP2-activated retGC activity in a manner similar to that by ATP pre-binding; however, amiloride did not directly stimulate the retGC activity. These results indicate that these two effects are mechanistically different. Levels of retGC activation by these effects and conditions required for these effects indicate that only the mechanism involving ATP pre-binding is physiologically relevant to retGC activation. PMID- 16259949 TI - Ginkgolides induce apoptosis and decrease cell numbers in mouse blastocysts. AB - In this report, we examine the cytotoxic effect of ginkgolides, the major components of Ginkgo biloba extracts, on the blastocyst stage of mouse embryos and on subsequent early postimplantation embryonic development in vitro. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling assay revealed that blastocysts treated with 5 or 10muM ginkgolide A or ginkgolide B showed increased apoptosis versus untreated controls. This could be correlated with the observation that ginkgolide-treated blastocysts showed a significant reduction in the average number of total cells in the blastocyst and trophectoderm/inner cell mass lineage versus controls. In addition, ginkgolide-pretreated blastocysts showed normal levels of implantation on culture dishes in vitro, but significantly fewer embryos reached the later stages of embryonic development in the treatment groups versus the controls, instead dying at relatively early stages of development. Our results collectively indicate that ginkgolide treatment of mouse blastocysts induces apoptosis, decreases cell numbers, retards early postimplantation blastocyst development, and increases early-stage blastocyst death. These novel findings provide important new insights into the effect of Ginkgo biloba extracts on mouse blastocysts. PMID- 16259950 TI - Transforming activity of the lymphotoxin-beta receptor revealed by expression screening. AB - Pancreatic ductal carcinoma (PDC) remains one of the most intractable human malignancies. To obtain insight into the molecular pathogenesis of PDC, we constructed a retroviral cDNA expression library with total RNA isolated from the PDC cell line MiaPaCa-2. Screening of this library with the use of a focus formation assay with NIH 3T3 mouse fibroblasts resulted in the identification of 13 independent genes with transforming activity. One of the cDNAs thus identified encodes an NH(2)-terminally truncated form of the lymphotoxin-beta receptor (LTBR). The transforming activity of this short-type LTBR in 3T3 cells was confirmed by both an in vitro assay of cell growth in soft agar and an in vivo assay of tumorigenicity in nude mice. The full-length (wild-type) LTBR protein was also found to manifest similar transforming activity. These observations suggest that LTBR, which belongs to the tumor necrosis factor receptor superfamily of proteins, may contribute to human carcinogenesis. PMID- 16259951 TI - Mitochondrial localization of mu-calpain. AB - Calcium-dependent cysteine proteases, calpains, have physiological roles in cell motility and differentiation but also play a pathological role following insult or disease. The ubiquitous calpains are widely considered to be cytosolic enzymes, although there has been speculation of a mitochondrial calpain. Within a highly enriched fraction of mitochondria obtained from rat cortex and SH-SY5Y human neuroblastoma cells, immunoblotting demonstrated enrichment of the 80kDa mu calpain large subunit and 28kDa small subunit. In rat cortex, antibodies against domains II and III of the large mu-calpain subunit also detected a 40kDa fragment, similar to the autolytic fragment generated following incubation of human erythrocyte mu-calpain with Ca(2+). Mitochondrial proteins including apoptosis inducing factor and mitochondrial Bax are calpain substrates, but the mechanism by which calpains gain access to these proteins is uncertain. Mitochondrial localization of mu-calpain places the enzyme in proximity to its mitochondrial substrates and to Ca(2+) released from mitochondrial stores. PMID- 16259952 TI - GATA4-mediated cardiac hypertrophy induced by d-myo-inositol 1,4,5-tris phosphate. AB - We evaluated the effects of d-myo-inositol 1,4,5-tris-phosphate on cardiac hypertrophy. d-myo-inositol 1,4,5-tris-phosphate augmented cardiac hypertrophy as evidenced by its effects on DNA synthesis, protein synthesis, and expression of immediate-early genes c-myc and c-fos, beta-myosin heavy chain, and alpha-actin. The administration of d-myo-inositol 1,4,5-tris-phosphate increased the expression of nuclear factor of activated T-cells and cardiac-restricted zinc finger transcription factor (GATA4). Real-time quantitative RT-PCR showed that d myo-inositol 1,4,5-tris-phosphate-induced GATA4 mRNA was significantly enhanced even in the presence of the calcineurin inhibitor, cyclosporine A. The effect of d-myo-inositol 1,4,5-tris-phosphate was blocked after inhibition of inositol trisphosphate receptors but not after inhibition of c-Raf/mitogen-activated protein kinase kinase (MEK)/mitogen-activated protein kinase (ERK) or p38 mitogen activated protein kinase pathways. The study shows that d-myo-inositol 1,4,5-tris phosphate-induced cardiac hypertrophy is mediated by GATA4 but independent from the calcineurin pathway. PMID- 16259953 TI - Nitric oxide synthase in the nervous system and ink gland of the cuttlefish Sepia officinalis: molecular cloning and expression. AB - Nitric oxide (NO) signaling is involved in numerous physiological processes in mollusks, e.g., learning and memory, feeding behavior, neural development, and defence response. We report the first molecular cloning of NOS mRNA from a cephalopod, the cuttlefish Sepia officinalis (SoNOS). SoNOS was cloned using a strategy that involves hybridization of degenerate PCR primers to highly conserved NOS regions, combined with RACE procedure. Two splicing variants of SoNOS, differing by 18 nucleotides, were found in the nervous system and the ink gland of Sepia. In situ hybridization shows that SoNOS is expressed in the immature and mature cells of the ink gland and in the regions of the nervous system that are related to the ink defence system. PMID- 16259954 TI - Identification of genes preferentially expressed in periodontal ligament: specific expression of a novel secreted protein, FDC-SP. AB - Gene expression in human periodontal ligament (PDL) was examined by suppression subtractive hybridization to identify genes that are preferentially expressed in tissue compared to cultured PDL fibroblasts. The most enriched genes in a subtracted cDNA library are primarily genes for extracellular matrix components, types I and III collagen, lumican, periostin, and asporin, among others, whose expression conveys unique mechanical properties to the PDL. Also within this group is the gene for follicular dendritic cell secreted protein (FDC-SP), a small protein like statherin in saliva, not previously found in PDL. FDC-SP's presence in PDL was confirmed by in situ hybridization in mouse which also showed that it was definitely present in the parotid gland, but, surprisingly, not in the other salivary glands: submandibular and sublingual. Since only normal tissue was examined, these findings suggest that FDC-SP plays an important but previously unsuspected role within oral connective tissue. PMID- 16259955 TI - Targeting of the mitochondrial membrane proteins to the cell surface for functional studies. AB - Studying mitochondrial membrane proteins for ion or substrate transport is technically difficult, as the organelles are hidden within the cell interior and thus inaccessible to many conventional nondisruptive techniques. This technical barrier can potentially be overcome if the mitochondrial membrane proteins are targeted to the cell surface, where they can be more readily studied. We undertook experiments presented here to target two related mitochondrial membrane proteins, mitochondrial ATP-binding cassette-1 and -2 protein (mABC1 and mABC2, respectively) to the cell surface for functional studies. These two proteins have an N-terminal mitochondrial targeting signal (MTS), and we hypothesized that removal of this sequence or addition of a cell surface targeting signal would lead to cell membrane targeting of these proteins. When the MTS was removed from mABC1, it localized to intracellular secretory compartments as well as the plasma membrane. However, truncated mABC2 lacking the MTS aggregated inside the cell. Addition of a cell membrane signal sequence or the transmembrane domain from CD8 to the N-terminus of mABC1 or mABC2 resulted in similar subcellular localizations. We then performed patch clamp on cells expressing mABC1 on their surface. These cells exhibited nonselective transport of K(+) and Na(+) ions and resulted in the loss of membrane potential. Our findings open new ways to study mitochondrial membrane proteins in established cell culture systems by targeting them to the cell surface, where they can more reliably be studied using various molecular and cellular techniques. PMID- 16259956 TI - Apolipoprotein A-I induces IL-10 and PGE2 production in human monocytes and inhibits dendritic cell differentiation and maturation. AB - Apolipoprotein A-I (apoA-I), the major protein component of serum high-density lipoprotein, exhibits anti-inflammatory activity in atherosclerosis. In this study, we demonstrate that apoA-I inhibits DC differentiation and maturation. DC differentiated from monocytes in the presence of apoA-I showed a decreased expression of surface molecules such as CD1a, CD80, CD86, and HLA-DR. In addition, these DC exhibited decreased endocytic activity and weakened allogeneic T-cell activation. During DC differentiation in the presence of apoA-I, PGE(2) and IL-10, which are known to be DC differentiation inhibitors and/or modulators of DC function, were produced at remarkable rates, whereas IL-12 production in the cells after stimulation with CD40 mAb and IFN-gamma was significantly decreased in comparison with the control DC. T cells stimulated by apoA-I pretreated DC produced significantly low levels of IFN-gamma, and apoA-I inhibited cross-talk between DC and NK cells, in terms of IL-12 and IFN-gamma production. Therefore, apoA-I appears to play an important role in modulating both innate immune response and inflammatory response. The novel inhibitory function of apoA-I on DC differentiation and function may facilitate the development of new therapeutic interventions in inflammatory diseases. PMID- 16259957 TI - Blocking the MyD88-dependent pathway protects the myocardium from ischemia/reperfusion injury in rat hearts. AB - We examined whether blocking the MyD88 mediated pathway could protect myocardium from ischemia/reperfusion (I/R) injury by transfecting Ad5-dnMyD88 into the myocardium of rats (n=8) 3 days before the hearts were subjected to ischemia (45min) and reperfusion (4h). Ad5-GFP served as control (n=8). One group of rats was (n=8) subjected to I/R without transfection. Transfection of Ad5-dnMyD88 significantly reduced infarct size by 53.6% compared with the I/R group (15.1+/ 3.02 vs 32.5+/-2.59) while transfection of Ad5-GFP did not affect I/R induced myocardial injury (35.4+/-2.59 vs 32.5+/-2.59). Transfection of Ad5-dnMyD88 significantly inhibited I/R-enhanced NFkappaB activity by 50% and increased the levels of phospho-Akt by 35.6% and BCL-2 by 81%, respectively. Cardiac myocyte apoptosis after I/R was significantly reduced by 59% in the Ad5-dnMyD88 group. The results demonstrate that both inhibition of the NFkappaB activation pathway and activation of the Akt signaling pathway may be responsible for the protective effect of transfection of dominant negative MyD88. PMID- 16259958 TI - Reactive oxygen species mediates homocysteine-induced mitochondrial biogenesis in human endothelial cells: modulation by antioxidants. AB - It has been proposed that homocysteine (Hcy)-induces endothelial dysfunction and atherosclerosis by generation of reactive oxygen species (ROS). A previous report has shown that Hcy promotes mitochondrial damage. Considering that oxidative stress can affect mitochondrial biogenesis, we hypothesized that Hcy-induced ROS in endothelial cells may lead to increased mitochondrial biogenesis. We found that Hcy-induced ROS (1.85-fold), leading to a NF-kappaB activation and increase the formation of 3-nitrotyrosine. Furthermore, expression of the mitochondrial biogenesis factors, nuclear respiratory factor-1 and mitochondrial transcription factor A, was significantly elevated in Hcy-treated cells. These changes were accompanied by increase in mitochondrial mass and higher mRNA and protein expression of the subunit III of cytochrome c oxidase. These effects were significantly prevented by pretreatment with the antioxidants, catechin and trolox. Taken together, our results suggest that ROS is an important mediator of mitochondrial biogenesis induced by Hcy, and that modulation of oxidative stress by antioxidants may protect against the adverse vascular effects of Hcy. PMID- 16259959 TI - Expression of Nanog gene promotes NIH3T3 cell proliferation. AB - Cells are the functional elements in tissue engineering and regenerative medicine. A large number of cells are usually needed for these purposes. However, there are numbers of limitations for in vitro cell proliferation. Nanog is an important self-renewal determinant in embryonic stem cells. However, it remains unknown whether Nanog will influence the cell cycle and cell proliferation of mature cells. In this study, we expressed Nanog in NIH3T3 cells and showed that expression of Nanog in NIH3T3 promoted cells to enter into S phase and enhanced cell proliferation. This suggests that Nanog gene might function in a similar fashion in mature cells as in ES cells. In addition, it may provide an approach for in vitro cell expansion. PMID- 16259961 TI - A plasminogen-like protein, present in the apical extracellular environment of thyroid epithelial cells, degrades thyroglobulin in vitro. AB - The prothyroid hormone, thyroglobulin (Tg), is stored at high concentrations in the thyroid follicular lumen as a soluble 19S homo-dimer and as heavier soluble (27S and 37S) and insoluble (Tgm) forms. Follicular degradation of Tg may contribute to maintaining Tg concentrations compatible with follicle integrity. Here, we report on the presence of a plasminogen-like protein in the follicular lumen of normal human thyroids and its synthesis and apical secretion by cultured epithelial thyroid cells. Since all the main luminal forms of Tg are cleaved by this plasminogen-like protein, we suggest that it contributes to Tg degradation in the follicular lumen. PMID- 16259960 TI - Role of growth hormone receptor signaling in osteogenesis from murine bone marrow progenitor cells. AB - Growth hormone (GH) regulates many of the factors responsible for controlling the development of bone marrow progenitor cells (BMPCs). The aim of this study was to elucidate the role of GH in osteogenic differentiation of BMPCs using GH receptor null mice (GHRKO). BMPCs from GHRKO and their wild-type (WT) littermates were quantified by flow cytometry and their osteogenic differentiation in vitro was determined by cell morphology, real-time RT-PCR, and biochemical analyses. We found that freshly harvested GHRKO marrow contains 3% CD34 (hematopoietic lineage), 43.5% CD45 (monocyte/macrophage lineage), and 2.5% CD106 positive (CFU F/BMPC) cells compared to 11.2%, 45%, and 3.4% positive cells for (WT) marrow cells, respectively. When cultured for 14 days under conditions suitable for CFU F expansion, GHRKO marrow cells lost CD34 positivity, and were markedly reduced for CD45, but 3- to 4-fold higher for CD106. While WT marrow cells also lost CD34 expression, they maintained CD45 and increased CD106 levels by 16-fold. When BMPCs from GHRKO mice were cultured under osteogenic conditions, they failed to elongate, in contrast to WT cells. Furthermore, GHRKO cultures expressed less alkaline phosphatase, contained less mineralized calcium, and displayed lower osteocalcin expression than WT cells. However, GHRKO cells displayed similar or higher expression of cbfa-1, collagen I, and osteopontin mRNA compared to WT. In conclusion, we show that GH has an effect on the proportions of hematopoietic and mesenchymal progenitor cells in the bone marrow, and that GH is essential for both the induction and later progression of osteogenesis. PMID- 16259962 TI - Expression profiles of various transporters for oligopeptides, amino acids and organic ions along the human digestive tract. AB - Various transporters such as H+/peptide cotransporter PEPT1 are expressed in the intestine, and play important physiological and pharmacological roles in the body. Present study was performed to examine the expression profile of 20 kinds of transporters (PEPT1 and 2, P-glycoprotein, amino acid transporters and organic ion transporters) along the human digestive tract, especially focusing on PEPT1. Using normal mucosal specimens, real-time polymerase chain reactions were carried out. Immunoblot analyses were also performed for PEPT1 expression. PEPT1 mRNA was highly expressed in the small intestine (duodenum>jejunum>ileum) compared to other tissues, and some patients showed a significant level of expression in the stomach. The expressional pattern of PEPT1 in the stomach and histological diagnosis indicated that gastric PEPT1 originated from the intestinal metaplasia. The amino acid transporters showed unique mRNA expression levels and distributions in the digestive tract. For example, the expression levels of B(0)AT1, a Na+-dependent and chloride-independent neutral amino acid transporter, were increased from the duodenum to ileum, which pattern is completely inverted to that for PEPT1. There is little expression of organic ion transporters except for organic cation/carnitine transporter OCTN2. In conclusion, PEPT1 was abundantly expressed in the small intestine, and the reciprocal expression of PEPT1 and B(0)AT1 may serve for the efficient absorption of protein digestive products. PMID- 16259963 TI - Specific adducts recognised by a monoclonal antibody against cisplatin-modified DNA. AB - Numerous clinical or experimental studies have employed monoclonal antibody CP9/19 for quantification of cisplatin DNA adducts. The nature of adducts recognised by CP9/19 on polymeric DNA were defined using synthetic deoxynucleotides reacted with cisplatin. Total adduct levels were determined by atomic absorption spectrometry. The nature of adducts formed were confirmed by analysis of enzymatic hydrolysates using an established ion-exchange chromatography method combined with inductively coupled plasma mass spectrometry. Of the Pt bound to oligonucleotide A (TTTTTGGTTTTTGGTTTTTGGTTTTTGGTTTTT), 77% was recovered in a product consistent with the expected 1,2 intra-strand cross-link between GG. For oligonucleotide B (TTTTTAGTTTTTAGTTTTTAGTTTTTAGTTTTT), 62% of the bound Pt was recovered in a product consistent with the 1,2 intra-strand cross link between AG. Of Pt bound to oligothymydylic acid, 65% was recovered in a product not previously described, small quantities of which were also formed on oligonucleotides A and B. The concentrations of adducts required to cause 50% reduction of signal in a competitive enzyme-linked immunosorbant assay (ELISA) (K values) were determined. Adducts on sequences containing no guanine or only non adjacent guanine residues, including sequences containing adenines adjacent to guanines, exhibited low or undetectable immunoreactivities (K-values = from 1 to >100 pmoles Pt per assay well). Adducts formed on oligodeoxynucleotides containing guanine doublets interspersed amongst thymine residues were the most immunoreactive (K-values: 2-7 fmoles adduct per assay well), comparable to adducts on calf-thymus DNA. The only cisplatin-DNA adducts recognised with high sensitivity by antibody CP9/19 were those involving adjacent guanine residues but immunorecognition of these was influenced by the surrounding DNA sequence. PMID- 16259964 TI - Inhibition of interferon-gamma signaling by a mercurio-substituted dihydropsoralen in murine keratinocytes. AB - Psoralens and ultraviolet light A (PUVA) are used in the treatment of a variety of epidermal proliferative and inflammatory disorders. These compounds are known to intercalate and photo crosslink DNA. Specific receptor proteins for psoralens have also been identified. We describe a novel activity of a thiol reactive derivative, iodomercurio-4',5'-dihydrotrimethylpsoralen (iodomercurio-H2TMP) in keratinocytes. Without UVA, this psoralen was found to be an effective inhibitor of interferon-gamma (IFN-gamma)-signaling as measured by induction of nitric oxide biosynthesis (IC50 = 0.8 microM). This activity was increased (IC50 = 0.1 microM) when the cells were depleted of intracellular glutathione (GSH) with buthionine sulfoximine. In keratinocytes, IFN-gamma stimulates expression of inducible nitric oxide synthase (NOS2). Although iodomercurio-H2TMP did not alter NOS2 enzymatic activity, it blocked IFN-gamma-induced expression of NOS2 mRNA and protein, an effect that was enhanced in GSH-depleted cells. Iodomercurio-H2TMP was found to readily inhibit IFN-gamma signaling in transient transfection assays using NOS2 promoter/luciferase reporter constructs. NOS2 gene expression is known to require a variety of transcription factors including STAT-1, NF-kappaB and AP 1. Using mobility shift assays the psoralen, at concentrations that inhibit nitric oxide biosynthesis, had no effect on the DNA binding activity of STAT-1 or NF-kappaB. However, iodomercurio-H2TMP was found to suppress AP-1. These data indicate that iodomercurio-H2TMP acts at sulfhydryl-sensitive sites to inhibit NOS2. Moreover, this is dependent on early events in the IFN-gamma signal transduction pathway. Inhibition of AP-1 suggests that the psoralen functions by interfering with an important transcription factor that regulates expression of NOS2 in keratinocytes. PMID- 16259965 TI - Effects of nicotine on hippocampal and cingulate activity during smooth pursuit eye movement in schizophrenia. AB - BACKGROUND: Abnormal smooth pursuit eye movement (SPEM) in schizophrenic patients is a well known phenomenon, but the neurophysiological mechanisms underlying the deficit are unknown. Nicotine temporarily improves SPEM and has been associated with reduced hippocampal hemodynamic activity in schizophrenics. Nicotine's effect on brain activity in control subjects performing SPEM has not been studied. The purpose of this work was to determine if nicotine differentially affects brain activity in schizophrenic and control subjects during pursuit eye tracking. METHODS: 16 subjects with schizophrenia and 16 control subjects underwent functional MR imaging during SPEM after receiving placebo or nicotine gum. Four brain regions were analyzed for main effects of group, drug, and interactions: hippocampus, cingulate gyrus, frontal eye fields, and area MT. RESULTS: Nicotine reduced hippocampal activity in both groups, but the effect was greater in control subjects. A group by drug interaction was observed in the anterior cingulate gyrus, where nicotine decreased activity in control subjects and increased activity in schizophrenic subjects. There were no significant effects of group, drug, or interactions in frontal eye fields or area MT. CONCLUSIONS: Nicotine may improve SPEM performance in people with schizophrenia through cholinergic stimulation of the hippocampus and cingulate gyrus. Potential mechanisms include improved inhibitory function and attention. PMID- 16259966 TI - IGF-1 protects oligodendrocyte progenitor cells and improves neurological functions following cerebral hypoxia-ischemia in the neonatal rat. AB - To investigate if insulin-like growth factor-1 (IGF-1) provides neuroprotection to oligodendrocyte progenitor cells (OPCs) following cerebral hypoxia-ischemia, a previously developed neonatal rat model of white matter damage was used in this study. Postnatal day 4 (P4) SD rat pups were subjected to bilateral common carotid artery ligation, followed by exposure to 8% oxygen for 10 min. IGF-1 (0.5 microg) or vehicle was injected into the left ventricle after artery ligation and before the hypoxic exposure. Cerebral hypoxia-ischemia caused death of O4+ late OPCs in the P5 rat brain and impaired myelination in the P9 and P21 rat brain. Caspase-3 activation was involved in the death of OPCs. Moreover, cerebral hypoxia-ischemia impaired neurobehavioral performance in juvenile rats. IGF-1 treatment attenuated damages to OPCs and improved neurological functions after cerebral hypoxia-ischemia. It reduced death of O4+ OPCs by 39% on P5 and enhanced myelination on P9 and P21. Bromodeoxyuridine uptake assay showed that cerebral hypoxia-ischemia inhibited proliferation of stem/progenitor cells in the subventricular zone and NG2+ early OPCs in the white matter area. IGF-1 treatment increased cell proliferation in the subventricular zone by 31% 1 day following hypoxic-ischemic insult. Proliferation of early and late OPCs in the IGF-1 treated group was 1.5- and 2.4-fold of that in the vehicle-treated group, respectively. In conclusion, IGF-1 provided potent neuroprotection to OPCs and improved neurological functions following cerebral hypoxia-ischemia in the neonatal rat. The neuroprotection of IGF-1 was associated with its antiapoptotic and mitogenic effects. PMID- 16259967 TI - Mu opioid receptor agonist DAMGO-induced suppression of saccharin intake in Lewis and Fischer rats. AB - Rats suppress intake of a saccharin cue when paired with a drug of abuse such as morphine or cocaine. Relative to Lewis rats, Fischer rats exhibit greater avoidance of a saccharin cue following saccharin-morphine pairings. The present study used the mu agonist, [D-Ala2,N-MePhe4,Gly-ol5]enkephalin (DAMGO), to test whether strain differences in sensitivity of the mu receptor contribute to this effect. Water-deprived Lewis and Fischer rats were given 5 min access to 0.15% saccharin followed by an icv injection of either DAMGO (0.5 microg/1 microl/rat) or an equal volume of saline. There were six taste-drug pairings occurring at 48 h intervals. The results showed that, relative to the saline treated controls, all rats reduced intake of the saccharin cue following saccharin-DAMGO pairings. No differences occurred between strains. These data suggest that greater morphine induced suppression of saccharin intake by the Fischer rats is not likely mediated by differences in sensitivity of the mu receptor. Other mechanisms are implicated. PMID- 16259968 TI - NMDA receptors mediate feeding elicited by neuropeptide Y in the lateral and perifornical hypothalamus. AB - Neuropeptide Y (NPY) and N-methyl-d-aspartate (NMDA) receptors in the lateral (LH) and perifornical hypothalamus (PFH) are believed to be involved in the stimulation of feeding behavior. To investigate the possibility that neurons with these receptors interact to stimulate eating, the NMDA receptor antagonists d-(-) 2-amino-5-phosphonopentanoic acid (D-AP5) or 7-chlorokynurenic acid (7-CK) were injected into the LH or PFH of satiated rats 5 min prior to NPY in the same site and subsequent food intake was measured 1, 2, and 4 h postinjection. The injection of NPY (78 pmol/0.3 microl aCSF) in the PFH produced an average food intake of 9.7 g in 4 h, compared to the intake of 1.3 g after the artificial cerebrospinal fluid (aCSF) vehicle. D-AP5 (1, 10, or 20 nmol/0.3 microl aCSF) pretreatment suppressed NPY-induced eating, with the 20 nmol dose of D-AP5 producing up to an 80% suppression of elicited food intake down to 1.9 g in 4 h. Similar effects were produced with the LH as the injection site. Illustrating the specificity of the NMDA receptor antagonist's suppression of NPY-elicited feeding, D-AP5 suppressed NMDA-elicited feeding but did not affect the eating response induced by kainic acid. Consistent with the effects of D-AP5, the NMDA receptor antagonist 7-CK (40 nmol/0.3 microl dimethyl sulfoxide, DMSO) suppressed feeding elicited by NPY in the LH by 78%. Collectively, the findings suggest that the feeding elicited by NPY is dependent upon the activation of the NMDA receptors in the LH and PFH. PMID- 16259969 TI - Effect of peripheral axotomy on pain-related behavior and dorsal root ganglion neurons excitability in NPY transgenic rats. AB - In order to clarify the physiologic role of NPY in sensory processing, we obtained intracellular recordings of DRG neurons from wild type (WT) and NPY overexpressing transgenic rats (NPY-TG) before and after injury. We investigated medium and large diameter DRG neurons since upregulation of NPY peptide following the nerve injury occurs primarily in those cells. Neurons were classified as Aalpha/beta and Adelta using conduction velocity and action potential duration. Prior to the injury, Aalpha/beta neurons of NPY-TG rats conducted more slowly and had a more brief AHP than similar cells from the WT group. Adelta neurons at baseline conducted faster in TG animals compared to WT. Ligation of the 5th lumbar spinal nerve (SNL) produced certain changes in Aalpha/beta cells that were evident only in the TG group. These include increased refractory period, increased input resistance, AHP prolongation and a depolarizing shift in threshold for AP initiation. The expected injury-induced CV slowing was not seen in NPY-TG Aalpha/beta cells. In the Adelta cell group, injury produced a depolarizing shift in the resting membrane potential, an increase in AP duration and decrease in AHP and refractory period duration only in WT rats, while NPY-TG cells lacked these injury-induced changes. Behavior tests showed diminished sensory response to nerve injury in NPY-TG rats, i.e., shorter duration of enhanced pain-related behavior and attenuation of contralateral effect. In conclusion, our observations suggest that NPY overexpression leads to reduced neuronal activity following nerve injury in a cell-specific manner. PMID- 16259970 TI - Enzymatic transglycosylation of xylose using a glycosynthase. AB - The application of the hyperactive glycosynthase derived from Agrobacterium sp. beta-glucosidase (AbgE358G-2F6) to the synthesis of xylo-oligosaccharides by using alpha-D-xylopyranosyl fluoride as donor represents the first successful use of glycosynthase technology for xylosyl transfer. Transfer to p-nitrophenyl beta D-glucopyranoside yields di- and trisaccharide products with beta-(1-->4) linkages in 63% and 35% yields, respectively. By contrast, transfer to p nitrophenyl beta-D-xylopyranoside yielded the beta-(1-->3) linked disaccharide and beta-D-Xyl-(1-->4)-beta-D-Xyl-(1-->3)-beta-D-Xyl-pNP as major products in 42% and 30% yields, respectively. Transfer of xylose to beta-D-Xyl-(1-->4)-beta-D-Xyl pNP yielded the beta-(1-->4) linked trisaccharide in 98% yield, thereby indicating that transfers to xylo-disaccharides occur with formation of beta-(1- >4) bonds. Xylosylation of carbamate-protected deoxyxylonojirimycin produced a mixture of di- and tri-'saccharide' products in modest yields. PMID- 16259971 TI - Structural insights into conformational flexibility at the peripheral site and within the active center gorge of AChE. AB - The peripheral anionic site on acetylcholinesterase (AChE), located at the active site gorge entry, encompasses overlapping binding sites for allosteric activators and inhibitors. Yet the molecular mechanisms coupling this site to the active center at the base of the gorge to modulate catalysis remain unclear. Crystal structures of mAChE bound with decidium, propidium and gallamine unveiled new determinants contributing to ligand interactions at the peripheral site. Subsequent studies using the syn and anti regioisomers of the click-chemistry inhibitor, TZ2PA6, that link propidium and tacrine moieties via distinctively substituted triazoles, revealed the inherent flexibility and a unique conformation of the peripheral site, along with substantial binding contributions from the triazoles with the Tyr337 region within the gorge. The recently solved structures of the mAChE mutant, Tyr337Ala, complexed with the TZ2PA6 isomers now reveals distinctive and time-dependent conformations of the complexes that are consistent with the triazole contribution to the energetics of inhibitor binding manifested in the respective dissociation rates of the complexes. PMID- 16259972 TI - Calculated low density lipoprotein cholesterol levels frequently underestimate directly measured low density lipoprotein cholesterol determinations in patients with serum triglyceride levels < or =4.52 mmol/l: an analysis comparing the LipiDirect magnetic LDL assay with the Friedewald calculation. AB - BACKGROUND: Increased low density lipoprotein cholesterol (LDL-C) is an established risk factor for the development of coronary artery disease (CAD). Recent guidelines detail specific LDL-C cutpoints for therapeutic goals. In practice, LDL-C is usually derived from the Friedewald formula (FF). This calculation is known to be inaccurate with serum triglyceride (TG) concentrations >4.52 mmol/l, however, its accuracy among relatively healthy patient cohorts with TG concentrations < or =4.52 mmol/l is less well studied. METHODS: We studied 661 ambulatory adults with TG concentrations < or =4.52 mmol/l and no overt CAD. Fasting venous lipid panels were obtained. LDL-C was calculated from the FF and also directly measured with the LipiDirect Magnetic LDL assay. Linear regression and paired t-test analyses were performed. RESULTS: Calculated and directly measured LDL-C concentrations were significantly different (4.26+/-0.88 vs. 4.83+/-1.06 mmol/l respectively, p<0.0001). In 93% of measurements directly measured LDL-C exceeded calculated LDL-C. Although calculated and directly measured LDL-C concentrations were related (R=0.90), the discrepancy between them increased linearly with increasing TG concentrations (R=0.67) and clinically important differences existed at normal or slightly increased TG concentrations. Concordant results for NCEP ATP-III risk categories were present for only 48.1% of samples. CONCLUSIONS: Significant differences between calculated and directly measured LDL-C using the LipiDirect Magnetic LDL assay exist in healthy subjects with TG < or =4.52 mmol/l. These differences are linearly related to TG concentrations and occur frequently at relatively low TG concentrations. PMID- 16259973 TI - Estimation of wavelet and short-time Fourier transform sonograms of normal and diabetic subjects' electrogastrogram. AB - Electrogastrography (EGG) is a noninvasive way to record gastric electrical activity of stomach muscle by placing electrodes on the abdominal skin. Our goal was to investigate the frequency of abnormalities of the EGG in real clinical diabetic gastroparesis patients using WT method and to compare performance of STFT and WT methods in the case of time-frequency resolution. The results showed that WT sonograms can be used to classify patients successfully as healthy or sick. And also, due to the fact that the WT method does not suffer from some intrinsic problems that affect the STFT method, one can see that the WT method can help improve the quality of the sonogram of the EGG signals. PMID- 16259974 TI - The Drosophila LIM-homeo domain protein Islet antagonizes pro-neural cell specification in the peripheral nervous system. AB - The pattern of the external sensory organs (SO) in Drosophila depends on the activity of the basic helix-loop-helix (bHLH) transcriptional activators Achaete/Scute (Ac/Sc) that are expressed in clusters of cells (pro-neural clusters) and provide the cells with the potential to develop a neural fate. In the mesothorax, the GATA1 transcription factor Pannier (Pnr), together with its cofactor Chip, activates ac/sc genes directly through binding to the dorso central enhancer (DC) of ac/sc. We identify the LIM-homeo domain (LIM-HD) transcription factor Islet (Isl) by genetic screening and investigate its role in the thoracic pre-patterning. We show that isl loss-of-function mutations result in expanded Ac expression in DC and scutellar (SC) pro-neural clusters and formation of ectopic sensory organs. Overexpression of Isl decreases pro-neural expression and suppresses bristle development. Moreover, Isl is coexpressed with Pnr in the posterior region of the mesothorax. In the DC pro-neural cluster, Isl antagonizes Pnr activity both by dimerization with the DNA-binding domain of Pnr and via competitive inhibition of the Chip-bHLH interaction. We propose that sensory organ pre-patterning relies on the antagonistic activity of individual Chip-binding factors. The differential affinities of these binding-factors and their precise stoichiometry are crucial in specifying pre-patterns within the different pro-neural clusters. PMID- 16259975 TI - The effects of olopatadine hydrochloride on the number of scratching induced by repeated application of oxazolone in mice. AB - It is suggested that atopic dermatitis is a skin disease associated with itching as subjective symptoms, and histamine H(1) receptor antagonists are used in order to prevent the itching, and the deterioration for scratch by itching. Histamine H(1) receptor selective anti-histamine olopatadine hydrochloride (olopatadine; Allelock shows consistent efficacy and safety in the treatment of allergic disorders. We investigated the possible efficacy of olopatadine on the number of scratching induced by repeated application of oxazolone in BALB/c mice. The repeated treatment of olopatadine significantly inhibited the ear swelling and the increased number of scratching. It significantly inhibited the increased production of interleukin (IL)-4, IL-1beta and granulocyte-macrophage colony stimulating factor (GM-CSF) in the lesioned ear. Moreover, it significantly inhibited the increased production of nerve growth factor (NGF) and substance P. On the other hand, loratadine, bepotastine and chlorpheniramine did not inhibit the ear swelling and the increased number of scratching. These results indicate that olopatadine inhibited not only the increased production of cytokines but also NGF and substance P unlike other histamine H(1) receptor antagonists. It was suggested that olopatadine suppressed the increased number of scratching by the anti-inflammatory effects. Therefore, olopatadine appears to exert additional biological effects besides its blockade of a histamine H(1) receptor. PMID- 16259976 TI - Costovertebral dysplasia in a patient with partial trisomy 22. AB - A newborn female presented with costovertebral dysplasia (CVD), subtle facial anomalies, and neonatal respiratory distress. Her karyotype demonstrated a small supernumerary NOR-positive marker that was subsequently identified as del(22)(q11.2). This extra structurally abnormal chromosome was found by DNA microsatellite marker analyses to be derived from a paternal chromosome 22. The child has had severe growth and developmental delay along with pulmonary insufficiency and hypoxia but is presently stable at age 20 months. Findings in our patient correlate with similar observations in children with small markers derived from D/G and D/D translocations reported before banding technology was available. These reports and recent mapping results suggest that a pericentric gene family, distributed on one or more acrocentric chromosomes, may have played a role in the development of the human axial skeleton. Data from additional studies will be needed to confirm or refute this hypothesis. PMID- 16259977 TI - Tenascin-C protein expression and mRNA splice variants in thyroid carcinoma. AB - Tenascin-C (Tn-C) is a matricellular protein involved in the initial and intermediate stages of cell adhesion. The present study is the first undertaken to comparatively investigate Tn-C in neoplastic, non-neoplastic thyroid lesions and normal thyroid tissues. Forty-eight thyroid specimens were studied immunohistochemically using a monoclonal antibody against Tn-C. Immunohistochemistry was supplemented by RT-PCR analysis of the two Tn-C mRNA splice variants in 13 thyroid cancer cell lines. Normal and non-neoplastic tissues were devoid of Tn-C, as well as follicular neoplasms, Huerthle-cell and anaplastic carcinomas. Most papillary carcinomas showed a focally intensive extracellular staining, localized in the connective tissue stroma, whereas most medullary carcinomas showed a staining in the connective tissue but also in intracellular location mainly. RT-PCR analysis detected Tn-C mRNA in all thyroid cancer cell lines with prevalence of the large splice variant in all but the medullary line, characterized by a higher Tn-Csmall:Tn-Clarge ratio. In conclusion, Tn-C re-expression has been observed in papillary and medullary thyroid carcinomas with different staining patterns accompanied by the prevalence of different mRNA splice variants in cell cultures. It seems possible that Tn-C is rather synthesized by tumor cells than by activated stromal cells. PMID- 16259978 TI - Rho kinase regulates fragmentation and phagocytosis of apoptotic cells. AB - During the execution phase of apoptosis, a cell undergoes cytoplasmic and nuclear changes that prepare it for death and phagocytosis. The end-point of the execution phase is condensation into a single apoptotic body or fragmentation into multiple apoptotic bodies. Fragmentation is thought to facilitate phagocytosis; however, mechanisms regulating fragmentation are unknown. An isoform of Rho kinase, ROCK-I, drives membrane blebbing through its activation of actin-myosin contraction; this raises the possibility that ROCK-I may regulate other execution phase events, such as cellular fragmentation. Here, we show that COS-7 cells fragment into a number of small apoptotic bodies during apoptosis; treating with ROCK inhibitors (Y-27632 or H-1152) prevents fragmentation. Latrunculin B and blebbistatin, drugs that interfere with actin-myosin contraction, also inhibit fragmentation. During apoptosis, ROCK-I is cleaved and activated by caspases, while ROCK-II is not activated, but rather translocates to a cytoskeletal fraction. siRNA knock-down of ROCK-I but not ROCK-II inhibits fragmentation of dying cells, consistent with ROCK-I being required for apoptotic fragmentation. Finally, cells dying in the presence of the ROCK inhibitor Y-27632 are not efficiently phagocytized. These data show that ROCK plays an essential role in fragmentation and phagocytosis of apoptotic cells. PMID- 16259979 TI - Smad7 mediates inhibition of Saos2 osteosarcoma cell differentiation by NFkappaB. AB - The transcription factor NFkappaB is constitutively activated in various tumor cells where it promotes proliferation and represses apoptosis. The bone morphogenetic proteins (BMPs) delay cell proliferation and promote differentiation and apoptosis of bone cells through activation of Smad downstream effectors and via Smad-independent mechanisms. Thus, NFkappaB and BMP pathways play opposing roles in regulating osteoblastic cell fate. Here, we show that in osteosarcoma Saos2 osteoblasts, NFkappaB regulates the activity of the BMP/Smad signaling. Inhibition of NFkappaB by overexpression of mIkappaB leads to the induction of osteoblast differentiation. Saos2 cells overexpressing mIkappaB (Saos2-mIkappaB) exhibit higher expression of osteoblast phenotypic genes such as alkaline phosphatase, Runx2 and osteocalcin and are more responsive to BMP2 in comparison to wild-type cells (Saos2-wt) or empty vector infected controls (Saos2 EV). Furthermore, BMP-2 signaling and Smad phosphorylation are significantly increased in Saos2-mIkappaB cells in comparison to Saos2-EV cells. Inhibition of NFkappaB signaling in Saos2-mIkappaB cells is associated with decreased expression of the BMP signaling inhibitor Smad7. While gain of Smad7 function in Saos2-mIkappaB cells results in inhibition of BMP signaling, anti-sense knockdown of Smad7 in Saos2-EV cells leads to upregulation of BMP signaling. We therefore conclude that in osteosarcoma Saos2 cells, NFkappaB represses BMP/Smad signaling and BMP2-induced differentiation through Smad7. PMID- 16259980 TI - Choroidal endothelial cells transmigrate across the retinal pigment epithelium but do not proliferate in response to soluble vascular endothelial growth factor. AB - The purpose of this study was to investigate the effects of soluble VEGF on human choroidal endothelial cell (CEC) transmigration across an RPE monolayer as it relates to choroidal neovascularization in AMD. In coculture assays, ARPE-19 (ARPE) was plated on the undersides of Transwell inserts having 0.4 microm pores. Primary human CECs were then plated into the insert. CECs in the Transwell inserts were counted after 72 hr of growth. CEC proliferation was also measured after culturing CECs in ARPE-CEC coculture-conditioned media or in media with exogenous VEGF121 and/or VEGF165 added. Transmigration assays were performed on Transwells with 8.0 microm pores: green-labelled CECs were plated in Transwell inserts with or without red-labelled ARPE plated on the undersides of the insert. In some transmigration assays, ARPE was plated into the wells to provide a chemotactic gradient for CEC transmigration. After 72 hr CECs were plated, green cells were counted either within the well media as CECs that transmigrated the epithelial monolayer, or on the underside of the insert as CECs that transmigrated the Transwell insert to but not beyond the ARPE monolayer. A neutralizing antibody to VEGF was added to the wells of Transwells at the time the CECs were plated in the insert and transmigrated CECs were counted. VEGF protein was measured in the conditioned media of ARPE and CEC coculture and in transmigration assays. Compared to control, CEC proliferation significantly increased when CECs were cultured in coculture conditioned media (p=0.001) or in coculture assays (p<0.001). However, there was no effect on CEC proliferation when VEGF121, VEGF165, or both were added to solo CECs. Antibody to VEGF did not reduce the proliferative effects of coculture conditioned media on CEC. ARPE plated in the well significantly increased CEC transmigration (p<0.001) compared to transmigration assays without ARPE in the well. VEGF protein measured in the well media of transmigration assays having ARPE within the well was significantly greater than in the assays without ARPE within the well (p<0.004). Exogenous neutralizing antibody to VEGF significantly reduced transmigration, and this effect was dose-dependent. VEGF provides a chemotactic gradient for human CECs to transmigrate across a monolayer of ARPE. Neutralization of VEGF in the media partially reduces transmigration. Whereas soluble VEGF does not increase proliferation of solo CECs, coculture conditioned media enhances proliferation, suggesting that growth factors other than VEGF cause CEC proliferation. These findings may have relevance to the transformation of occult CNV into CNV within the neurosensory retina in AMD. PMID- 16259981 TI - Physiologic progesterone reduces mitochondrial dysfunction and hippocampal cell loss after traumatic brain injury in female rats. AB - Growing literature suggests important sex-based differences in outcome following traumatic brain injury (TBI) in animals and humans. Progesterone has emerged as a key hormone involved in many potential neuroprotective pathways after acute brain injury and may be responsible for some of these differences. Many studies have utilized supraphysiologic levels of post-traumatic progesterone to reverse pathologic processes after TBI, but few studies have focused on the role of endogenous physiologic levels of progesterone in neuroprotection. We hypothesized that progesterone at physiologic serum levels would be neuroprotective in female rats after TBI and that progesterone would reverse early mitochondrial dysfunction seen in this model. Female, Sprague-Dawley rats were ovariectomized and implanted with silastic capsules containing either low or high physiologic range progesterone at 7 days prior to TBI. Control rats received ovariectomy with implants containing no hormone. Rats underwent controlled cortical impact to the left parietotemporal cortex and were evaluated for evidence of early mitochondrial dysfunction (1 h) and delayed hippocampal neuronal injury and cortical tissue loss (7 days) after injury. Progesterone in the low physiologic range reversed the early postinjury alterations seen in mitochondrial respiration and reduced hippocampal neuronal loss in both the CA1 and CA3 subfields. Progesterone in the high physiologic range had a more limited pattern of hippocampal neuronal preservation in the CA3 region only. Neither progesterone dose significantly reduced cortical tissue loss. These findings have implications in understanding the sex-based differences in outcome following acute brain injury. PMID- 16259982 TI - Lesional accumulation of P2X4 receptor+ monocytes following experimental traumatic brain injury. AB - P2X4 receptor (P2X4R) is an ATP-gated ion channel. ATP is an important messenger in traumatic brain injury. Here, we report expression of P2X4R in rat traumatic brain injury with focus on the early phase, most amenable to therapy. Accumulation of P2X4R+ cells was observed as early as 6 h after injury and continued to increase 4 days post-injury at the lesion and remote areas. Double staining revealed that most P2X4R+ cells co-expressed ED-1, a marker for reactive microglia/macrophages, but not nestin or W3/13. Our data suggest that P2X4R expression defines a subtype of activated microglia/macrophages involved in the early processes following traumatic brain injury. PMID- 16259983 TI - Minocycline and intracerebral hemorrhage: influence of injury severity and delay to treatment. AB - Intracerebral hemorrhage (ICH) is a devastating condition currently lacking a defined line of treatment. The inflammatory response that ensues following its onset is thought to contribute to secondary injury following ICH, making inflammation a potential therapeutic target. Minocycline (MC), a commonly used antibiotic that also has anti-inflammatory and anti-apoptotic properties, provides histological protection in several animal stroke models when given soon after injury. However, its ability to provide protection with more clinically relevant delays is unknown. The objective of this study was to examine the effects of MC on histopathological changes and long-term functional outcomes in a collagenase-induced ICH model in rats when drug administration was delayed 3 h following the onset of ICH. In accordance with other studies, MC suppressed microglial/macrophage activation in the peri-infarct region at 5 days based on B4 isolectin histochemistry. However, no reduction in infarct volume was detected at 5 or 28 days post-ICH. Minocycline given for either 5 or 14 days also provided no functional benefit as assessed with a battery of sensory-motor tests (i.e., staircase, cylinder, ladder tests). These findings raise questions about the ability of MC to provide protection in ICH when delay to treatment is increased. PMID- 16259984 TI - Production and purification of recombinant somatolactin beta and its effects on melanosome aggregation in zebrafish. AB - A second form of somatolactin, somatolactin beta (SLbeta), was recently discovered in zebrafish (Danio rerio). This novel subtype of somatolactin is distantly related to somatolactin alpha (SLalpha) found in teleost species and is produced in a different region of the pituitary. To date, no physiological study of SLbeta has been reported. In order to study the physiological functions of SLbeta, recombinant SLbeta protein has been produced and purified. The cDNA of zebrafish SLbeta was cloned into a pET100 bacteria expression vector and His tagged fusion proteins were produced in BL21 (DE3) Escherichia coli cells. The majority of recombinant somatolactins produced by E. coli were isolated in inclusion bodies although a small percentage of recombinant proteins (<1%) were also found in soluble form. Fusion proteins were solubilized from inclusion bodies using 6M guanidine hydrochloride. Pure recombinant somatolactins were obtained by affinity purification. The estimated molecular weight of 28 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis correlates with the molecular mass calculated from the deduced amino acid sequence of SLbeta. Thereafter, specific polyclonal antibodies against the recombinant SLbeta were developed. These antibodies recognized specifically a group of cells located in the anterior pars intermedia of the pituitary. The antibodies did not react with SLalpha, growth hormone or prolactin cells in the zebrafish pituitary glands. Furthermore, recombinant SLbeta induced melanosome aggregation in a concentration dependent manner in skin of zebrafish scales. Significant melanosome aggregation was observed in zebrafish melanophores at a concentration of 1 microg/ml. These results, combined with previous reports demonstrate that the recombinant SLbeta proteins produced here are bioactive. The function of inducing melanosome aggregation is conserved among the somatolactin functions. PMID- 16259985 TI - Meet the (burying) beetles. PMID- 16259986 TI - Evolving trends in the care of polytrauma patients. AB - Management of polytrauma patients has changed considerably in recent years. This is in keeping with the developments that have occurred in the fields of fracture fixation techniques and intensive care. Prior to the 1970s, patients with multiple injuries were treated non-operatively, as it was believed that they were too ill to withstand surgery. Around this time, literature appeared to suggest that these patients had high rates of complications as a result of prolonged recumbency. Fracture fixation techniques were also developing rapidly, and these events led to the advent of early fracture stabilisation of multiply injured patients, known as early total care. In the following decade, the surgical world came to recognise that early stabilisation of skeletal injuries produced poor results in certain patients. The concept of 'damage control' surgery was introduced for multiply injured patients. The current era may give way to new methods as our understanding of the pathophysiology of polytrauma improves. PMID- 16259987 TI - Risk factors and spatial patterns of hookworm infection among schoolchildren in a rural area of western Cote d'Ivoire. AB - This study is aimed at investigating the risk factors for hookworm infection among schoolchildren in a rural area of western Cote d'Ivoire and predicting and mapping the spatial distribution of infection. We used demographic and socio economic data from a cross-sectional survey of 6-16-year-old schoolchildren from 56 schools. Infection with hookworm was determined by microscopic examination of stool samples employing the Kato-Katz technique and an ether-concentration method. Environmental data were derived from satellite images and digitised maps. Bayesian variogram models were applied to investigate the variation of hookworm infection in relation to demographic, socio-economic and environmental factors. The overall hookworm infection prevalence, based on the pooled microscopic diagnoses, was 43.3% and ranged from 5.4 to 79.1% in the schools surveyed. Bivariate analyses showed that sex, age, socio-economic status, elevation, rainfall and land cover were significantly associated with the spatial distribution of hookworm infection. The final multivariate spatial model consisted of the covariates age, sex, socio-economic status, elevation and land cover. When assuming non-stationary underlying spatial dependency, the results of the model suggested that spatial correlation depended on the location only marginally. We conclude that, at the current resolution, it seems more reasonable to target interventions based on well-established epidemiologic risk factors, rather than on spatial factors. PMID- 16259989 TI - Finite element modeling of a progressively expanding shape memory stent. AB - Cardiovascular stents are small cylindrical devices introduced in stenosed arteries to reopen the lumen and restore blood flow. However, this treatment presents complications, including restenosis, which is the reclosing of the artery's diameter after the insertion of a stent. The structure of the prosthesis penetrates into and injures the walls of the patient's artery. There then follows a proliferation of cells and the formation of scar tissue around the injury, similar to the scarring of other organic tissues. This reaction to the trauma subjects the artery to close. The proposed solution is to develop a Nitinol stent with a progressive expansion device made of polyethylene, allowing smooth and gradual contact between the stent and the artery's wall by creep effect. The purpose of this paper is to describe the technology and methodology for the numerical study of this kind of stent through the finite element method. ANSYS 8.0 software is used to perform the analysis. The Nitinol is modeled with a superelastic law and the polyethylene with a yield hardening law. A first simulation determines the final geometry of the stent laser cut from a small tube. A second simulation examines the behavior of the prosthesis during surgery and over the 4 weeks following the operation. The results demonstrate that a compromise can be reached between a limited expansion prior the inflation of the expandable balloon and a significant expansion by creep of the polymer rings. PMID- 16259988 TI - Intraplaque MMP-8 levels are increased in asymptomatic patients with carotid plaque progression on ultrasound. AB - Carotid atherosclerotic plaque remodelling and increased risk of symptomatic plaque rupture seem to be partially mediated by matrix metalloproteinases (MMPs). In this study, we have investigated whether different MMPs are related to carotid atherosclerosis or to recent ischaemic brain disease. Eighty-four consecutive patients undergoing carotid endarterectomy for symptomatic and asymptomatic disease were studied. Plaques were analysed by ultrasound and later by morphology. Plasma MMP-2, MMP-8 and MMP-9 levels were quantified by ELISA. MMP expression and activity in carotid plaques was analysed by Western blotting and in situ zymography. Results were analysed with respect to plaque stability, morphology, symptomatic disease, presence of vascular risk factors and plasma markers of acute inflammation as high sensitivity C-reactive protein (hsCRP), fibrinogen, D-dimer and white blood cell counts. Patients with hypoechogenic plaques on ultrasound had more plasma MMP-8 (p = 0.04) and increased MMP activity as assessed by in situ zymography. Asymptomatic patients with plaque progression had more active intraplaque MMP-8 than asymptomatic patients without plaque progression. Presence of recent intraplaque haemorrhage or past history of CAD was related to increased activity of MMPs as assessed by in situ zymography (p < 0.01, CI 95% 0.8-1.0). Plasma MMP-8 and MMP-9, but not MMP-2 levels, decrease with time after ischaemic stroke. Patients with hypertension had more intraplaque active MMP-9 than normotensive (p = 0.03, CI 95% 0.7-1.0). Hypoechogenic carotid plaques had increased MMP activity and asymptomatic patients with plaque progression show increase intraplaque MMP-8 levels. PMID- 16259991 TI - Estimation of the axis of a screw motion from noisy data--a new method based on Plucker lines. AB - The problems of estimating the motion and orientation parameters of a body segment from two n point-set patterns are analyzed using the Plucker coordinates of a line (Plucker lines). The aim is to find algorithms less complex than those in conventional use, and thus facilitating more accurate computation of the unknown parameters. All conventional techniques use point transformation to calculate the screw axis. In this paper, we present a novel technique that directly estimates the axis of a screw motion as a Plucker line. The Plucker line can be transformed via the dual-number coordinate transformation matrix. This method is compared with Schwartz and Rozumalski [2005. A new method for estimating joint parameters from motion data. Journal of Biomechanics 38, 107 116] in simulations of random measurement errors and systematic skin movements. Simulation results indicate that the methods based on Plucker lines (Plucker line method) are superior in terms of extremely good results in the determination of the screw axis direction and position as well as a concise derivation of mathematical statements. This investigation yielded practical results, which can be used to locate the axis of a screw motion in a noisy environment. Developing the dual transformation matrix (DTM) from noisy data and determining the screw axis from a given DTM is done in a manner analogous to that for handling simple rotations. A more robust approach to solve for the dual vector associated with DTM is also addressed by using the eigenvector and the singular value decomposition. PMID- 16259993 TI - Evaluation of solid-phase microextraction as an alternative to the official method for the analysis of organic micro-pollutants in drinking water. AB - The objective this study was to compare the official EU liquid-liquid extraction (LLE) method with solid-phase microextraction (SPME) for the analysis of compounds migrating from cross-linked polyethylene into water. A medium polarity polydimethylsiloxane/divinylbenzene (PDMS/DVB) 65 microm fibre proved most efficient for the SPME extraction of nine test compounds and the optimum extraction conditions were an immersion time of 30 min with heating to 60 degrees C. The repeatability of the SPME method was variable: RSD values ranged from approximately 4-18% depending on the individual compound, though correlation coefficients were greater than 0.999 in the concentration range 0.5-1000 microg/l. It would also seem that there is some competition amongst different compounds for sites on the fibre and this is a potential drawback of SPME when applied to unknown samples. However, when applied to water samples in contact with polyethylene, SPME proved to be immensely more sensitive and to have a greater extraction range than LLE. These factors coupled with the rapidity and ease of use of SPME mean that it could be developed for use as an alternative to the existing official method or as an alert system in the routine analysis of materials used to transport domestic water. PMID- 16259992 TI - Solid phase extraction of some precious metals from hydrochloric acid to polystyrene-divinylbenzene porous resin impregnated with polyoxyethylene-type nonionic surfactant. AB - The solid phase extraction of gold(III), platinum(II), and palladium(II) to surfactant-impregnated polystyrene-divinylbenzene porous resin (XAD-4) was studied. The extracting media could be prepared just by mixing the resin in aqueous surfactant solutions. XAD-4 impregnated with a nonionic surfactant, polyethylene glycol monooleyl ether, was useful for extracting gold(III) from hydrochloric acid. The extractions of platinum(II) and palladium(II) were improved in the use of XAD-4 impregnated with a nitrogen-containing nonionic surfactant, polyethylene glycol stearyl amine. On the other hand, base metals such as copper(II), cobalt(II), nickel(II) and zinc(II), were hardly extracted. PMID- 16259994 TI - Wetting properties at the surface of iota-carrageenan-based edible films. AB - Surface properties of edible films composed of a polymeric matrix of carrageenan in association with hydrophobic material were studied by contact angle measurements. The use of this technique not only in a static mode but also in a dynamic way enables investigation of surface hydrophobicity as well as surface wettability. The absorption flux inside the material can be estimated from the wetting kinetic, which can be very useful to quickly compare water barrier efficiency of the tested films. Comparison of carrageenan films with films containing known amounts of additives enables understanding and correlation of changes of the surface properties with the nature of used additives (glycerol used as a plasticizer, glycerol monostearate used as a surfactant, and fat) and their influence on the orientation of polymer chains at the surface during film formation. Very different responses were observed from one surface of the film (film-casting-support interface) to the other (film-air interface), which could be also attributed to the influence of the support on the polymer and to macromolecular orientation during drying after casting. PMID- 16259995 TI - Effect of stress-jump condition on electrophoretic behavior of a spherical dispersion of soft particles. AB - The electrophoresis of a concentrated dispersion of soft particles, where a particle comprises a rigid core and an ion-penetrable membrane layer, is modeled theoretically, taking the effect of double-layer polarization into account. In particular, the influence of a stress-jump condition of the flow field at the membrane layer-liquid interface on the electrophoretic mobility of a particle is investigated. The type of particles considered mimic biocolloids, such as cells and microorganisms, and inorganic colloids covered by an artificial polymer layer such as surfactant molecules. A unit cell model is adopted to simulate the present spherical dispersion, and the governing equations and the associated boundary conditions are solved by a pseudo-spectral method based on Chebyshev polynomials. We show that while the stress-jump condition, characterized by a stress-jump coefficient, can have a significant influence on the mobility of a particle, the associated flow field is not influenced appreciably. Also, the influence of the stress-jump condition on the mobility of a particle depends largely on the nature of the membrane layer, characterized by its friction coefficient. PMID- 16259996 TI - Synthesis and characterization of new cationic quaternary ammonium polymerizable surfactants. AB - Two new polymerizable surfactants (surfmers), (11 acryloyloxyundecyl)dimethylethylammonium bromide (ethyl surfmer) and (11 acryloyloxyundecyl)dimethyl(2-hydroxyethyl)ammonium bromide (hydroxyethyl surfmer), were synthesized and characterized. The binary phase diagrams of both surfmer/water systems are described. Both surfmers can form isotropic solutions and lamellar lyotropic liquid crystalline phases. The hydroxyethyl surfmer/water system forms a lamellar phase for weight concentrations of surfmer between 70 and 90% relative to between 75 and 85% for the ethyl surfmer/water system. The differences in the self assembly of these surfmers were attributed to the ability of hydroxyethyl surfmer to form hydrogen bonds (between two head groups and with water), whereas no such interactions can occur with the ethyl surfmer system. PMID- 16259997 TI - A stable disulfide-free gene-3-protein of phage fd generated by in vitro evolution. AB - Disulfide bonds provide major contributions to the conformational stability of proteins, and their cleavage often leads to unfolding. The gene-3-protein of the filamentous phage fd contains two disulfides in its N1 domain and one in its N2 domain, and these three disulfide bonds are essential for the stability of this protein. Here, we employed in vitro evolution to generate a disulfide-free variant of the N1-N2 protein with a high conformational stability. The gene-3 protein is essential for the phage infectivity, and we exploited this requirement for a proteolytic selection of stabilized protein variants from phage libraries. First, optimal replacements for individual disulfide bonds were identified in libraries, in which the corresponding cysteine codons were randomized. Then stabilizing amino acid replacements at non-cysteine positions were selected from libraries that were created by error-prone PCR. This stepwise procedure led to variants of N1-N2 that are devoid of all three disulfide bonds but stable and functional. The best variant without disulfide bonds showed a much higher conformational stability than the disulfide-containing wild-type form of the gene 3-protein. Despite the loss of all three disulfide bonds, the midpoints of the thermal transitions were increased from 48.5 degrees C to 67.0 degrees C for the N2 domain and from 60.0 degrees C to 78.7 degrees C for the N1 domain. The major loss in conformational stability caused by the removal of the disulfides was thus over-compensated by strongly improved non-covalent interactions. The stabilized variants were less infectious than the wild-type protein, probably because the domain mobility was reduced. Only a small fraction of the sequence space could be accessed by using libraries created by error-prone PCR, but still many strongly stabilized variants could be identified. This is encouraging and indicates that proteins can be stabilized by mutations in many different ways. PMID- 16259998 TI - Striatal dopamine release after prefrontal repetitive transcranial magnetic stimulation in major depression: preliminary results of a dynamic [123I] IBZM SPECT study. AB - Though there is considerable evidence that prefrontal repetitive transcranial magnetic stimulation (rTMS) exerts antidepressant effects, the neurobiological action of rTMS in patients with depression is poorly understood. Preclinical studies in animals and humans have demonstrated that prefrontal rTMS can induce dopamine release in mesostriatal and mesolimbic regions. We therefore investigated whether rTMS also modulates striatal dopaminergic neurotransmission in depressed patients using a dynamic [123I] iodobenzamide (IBZM) single photon emission computed tomography (SPECT) approach. Five patients with a major depressive episode (DSM-IV) underwent an acute 10 Hz rTMS challenge with 3000 stimuli over the left dorsolateral prefrontal cortex during an [123I] IBZM-SPECT bolus and constant infusion protocol. In four subjects the protocol was repeated after a three week rTMS standard treatment. Striatal IBZM binding to dopamine D2 receptors was assessed with a region-of-interest (ROI) technique. The change in striatal IBZM binding after the rTMS challenge was regarded as measure of change in endogenous striatal dopamine. Data of nine SPECT investigations showed a significant reduction by 9.6+/-6.2% in IBZM binding to striatal dopamine D2 receptors after rTMS challenge compared to baseline (p=0.01, Wilcoxon test). In this preliminary study, the reduction of IBZM binding observed after rTMS challenge is suggestive of a release in endogenous dopamine induced by prefrontal rTMS. In future, this approach can be used to differentiate specific and non specific reward-related effects of rTMS on dopaminergic neurotransmission. PMID- 16259999 TI - The herbal medicine, Dai-Kenchu-to, accelerates delayed gastrointestinal transit after the operation in rats. AB - BACKGROUND: Post-operative ileus (POI) is a transient bowel dysmotility after operation. We have previously shown that laparotomy alone significantly delayed gastrointestinal (GI) transit, compared to anesthesia alone. The GI transit was further delayed after laparotomy plus intestinal manipulation. Dai-Kenchu-to (DKT), an herbal medicine, has been used for treating adhesive bowel obstruction in Japan. We studied whether DKT improves delayed GI transit after the operation, with or without morphine administration in rats. MATERIALS AND METHODS: Under isoflurane anesthesia, POI was induced by laparotomy with intestinal manipulation. Immediately after the operation, the rats received 51Cr by gavage. Three hours after the operation, the rats were sacrificed and GI transit was estimated by calculating the geometric center (GC). DKT (120, 360, and 1,200 mg/kg) were administered by gavage after the operation, with or without morphine administration (1 mg/kg s.c.). A muscarinic receptor antagonist (atropine; 50 mug/kg), a 5HT3 receptor antagonist (ondansetron; 1 mg/kg) and a 5HT4 receptor antagonist (GR113,808; 3 mg/kg) were administered before the operation. Truncal vagotomy was performed preceding the operation. RESULTS: Laparotomy with intestinal manipulation produced a significant delay in GI transit (GC = 2.93 +/- 0.16), compared to that of anesthesia alone (9.51 +/- 0.45). DKT at the dose of 360 mg/kg (GC = 3.77 +/- 0.10, P < 0.01) and 1,200 mg/kg (GC = 3.77 +/- 0.20, P < 0.01) significantly accelerated delayed GI transit induced by operation. Ondansetron, GR113,808, atropine, and truncal vagotomy abolished the stimulatory effect of DKT (360 mg/kg). When morphine was administered, GI transit was further reduced (GC = 1.97 +/- 0.10). DKT at the dose of 360 mg/kg (GC = 2.81 +/- 0.22, P < 0.05) and 1,200 mg/kg (GC = 2.87 +/- 0.23, P < 0.05) significantly improved delayed GI transit in morphine treated rats. CONCLUSIONS: DKT accelerates delayed GI transit induced by intestinal manipulation with and without concomitant morphine administration. DKT treatment may be useful for the patients with POI. PMID- 16260000 TI - Tissue distribution and induction of the rat multidrug resistance-associated proteins 5 and 6. AB - Multidrug resistance-associated proteins (Mrps) are ATP-dependent transporters which transport a wide variety of anionic and cationic compounds. The purpose of this study was to determine the tissue distribution of Mrp5 and 6 in male and female Sprague-Dawley rats in various tissues, and to investigate whether the expression is altered by cholestasis or administration of microsomal enzyme inducers (MEIs). These MEIs activate six different transcriptionally-mediated pathways, and their effects on Mrp5 and Mrp6 expression were determined. The effects of bile-duct ligation, a cholestasis model, on Mrp5 and 6 expression in male rats were quantified. Mrp5 had marked expression in adrenal gland, and moderate expression in cerebral cortex, cerebellum, and stomach. The MEIs polychlorinated biphenyl (PCB)126, phenobarbital, and PCB99 slightly repressed Mrp5, but no single class of receptor agonists induced or repressed Mrp5. Bile duct ligation tended to increase Mrp5 expression, but was not statistically significant at a 3 day timepoint. Mrp6 expression was highest in intestine, liver, and kidney. Mrp6 was slightly repressed by phenobarbital, dexamethasone, and isoniazid, but no one class of receptor agonists induced or repressed Mrp6, and expression was also unchanged bile-duct ligation. In conclusion, Mrp5 in rats is most highly expressed in the adrenal gland, whereas Mrp6 is mainly expressed in excretory organs (liver, intestine, and kidney), suggesting markedly different functions. Hepatic mRNA levels of Mrp5 or Mrp6 do not seem to be coordinately regulated along with Phase I enzymes via receptor-mediated pathways, and are not part of the hepatoprotective upregulation of basolateral transporters that occurs during cholestasis. PMID- 16260001 TI - Stevioside does not cause increased basal insulin secretion or beta-cell desensitization as does the sulphonylurea, glibenclamide: studies in vitro. AB - We have shown that stevioside (SVS) enhances insulin secretion and thus may have a potential role as antihyperglycemic agent in the treatment of type 2 diabetes mellitus. However, whether SVS stimulates basal insulin secretion (BIS) and/or cause desensitization of beta cells like sulphonylureas (SU), e.g. glibenclamide (GB), is not known. To explore and compare the effects of SVS pretreatment with those of GB and glucagon-like peptide-1 (GLP-1), we exposed isolated mouse islets to low or high glucose for 1 h after short-term (2 h) or long-term (24 h) pretreatment with SVS, GB or GLP-1, respectively. BIS at 3.3 or 5.5 mM glucose were not changed after short-term pretreatment with SVS (10(-7) M), while it increased about three folds after pretreatment with GB (10(-7) M). Glucose stimulated insulin secretion (GSIS) (16.7 mM) increased dose-dependently after long-term pretreatment with SVS at concentrations from 10(-7) to 10(-5) M. Pretreatment for 24 h with GB (10(-7) M) increased the subsequent BIS (3.3 mM glucose) (p < 0.001), but decreased GSIS (16.7 mM glucose) (p < 0.001). In contrast SVS (10(-7) M) and GLP-1 (10(-7) M) did not stimulate BIS but both enhanced the subsequent GSIS (16.7 mM glucose) (p < 0.05 and p < 0.05, respectively). While SVS pretreatment increased the intracellular insulin content, GB pretreatment decreased the insulin content. Our study suggests that SVS pretreatment does not cause a stimulation of BIS and does not desensitize beta-cells, i.e. SVS seems to have advantageous characteristics to GB as a potential treatment of type 2 diabetes. PMID- 16260002 TI - Apoptosis induction by dohevanil, a DHA substitutive analog of capsaicin, in MCF 7 cells. AB - Capsaicin (8-methyl-N-vanillyl-6-nonenamide), a major pungent ingredient in a variety of red peppers of the genus Capsicum, is a type of vanilloid. It has been shown to induce apoptosis in many cell types. The effects of vanilloids on apoptosis induction are thought to be correlated with the length and degree of the unsaturation of the fatty acyl chains. In this study, we compared the effect of capsaicin and its docosahexaenoic acid (DHA, C22:6) analog (we named as dohevanil) on human breast cancer MCF-7 cells, which do not express caspase-3. Dohevanil, which was synthesized from DHA and vanillylamine, has longer and highly unsaturated fatty acyl chain than capsaicin. We showed that both vanilloids exhibit effects of growth inhibition and DNA fragmentation induction in MCF-7 cells. These effects of dohevanil were more potent than capsaicin. Because these effects were inhibited by z-VAD-fmk, a broad-spectrum caspase inhibitor, the vanilloids induced the apoptosis via caspase-dependent pathway not involving caspase-3. In conclusion, dohevanil has a more potent effect on apoptosis induction in MCF-7 cells than capsaicin. PMID- 16260003 TI - Blood volume measurement: The comparison of pulse dye densitometry and Dill and Costill's methods. AB - In many clinical situations, it is crucial to determine circulating blood volume (BV) easily and to repeat this measurement. The Dye DensitoGram Analyzer (DDG, Nihon Kohden Corp) measures semi-automatically BV, using an injection of IndoCyanine Green (ICG, 10 mg), and avoiding intermittent blood samples. The DDG was used during a 90-day microgravity simulation by Head-Down-Tilt bed rest (HDT) to measure BV and compared with the calculation of the plasma volume (PV) variations according to Dill and Costill's formula (DC). Seventeen healthy volunteers were included: 8 control subjects (Co) and 9 subjects submitted to a resistive exercise counter-measure (CM). Measurements were performed, one day before HDT, on days 3 and 90 of HDT and on day 9 after HDT. A double measurement of the BV was performed to assess the repeatability of this method. On the last day of HDT a significant decrease (p < 0.05) in the PV was noted with the DDG (Co: -12.3+/-5.7%, CM: -9.0+/-5.3%) and DC; (Co: -4.7+/-1.8%, CM: -6.8+/-2.5%). A good repeatability of the technique was shown with a low intrasubjects coefficient of variation (4.95+/-0.95%) and an acceptable intersubjects coefficient of variation (15.30+/-1.13%). No correlation was noted between DDG and DC (r2 = 0.27). The DDG gives a good repeatability, not affected by the microgravity exposure. Thanks to its capacity to measure accurately the BV within 7-10 min, this device presents major advantages for clinical use and research purpose. PMID- 16260004 TI - Amphipod acute and chronic sediment toxicity assessment in estuarine environmental monitoring: An example from Ria de Aveiro, NW Portugal. AB - This paper presents a comprehensive estuarine sediment toxicity survey, using acute and full life-cycle tests with an indigenous species, the amphipod Corophium multisetosum. In the acute test, adult specimens were exposed to sediments from 144 sites for 10 days. In the full life-cycle test, 5-7 days old specimens, obtained in laboratory cultures, were exposed during 21 days to sediments from 56 sites. Three endpoints were considered: survival, for the acute and the chronic test and fecundity (number of gravid females per replicate at the end of the exposure) and growth (weight increase), for the chronic test. In the acute tests, adult survival was high overall and almost no stressed areas could be recognized in the estuary. This image was not reflected by the chronic endpoints, in particular growth and fecundity, which, corrected for the number of survivors, was the most affected descriptor. The chronic data could not be related to the baseline sediment variables analysed (grain-size, total organic content and redox potential), suggesting that these play no part in the biological responses. Results indicate that a tiered sediment bioassessment approach including an initial comprehensive ecotoxicological screening phase should not rely solely upon acute responses, although these are obtained from simpler and more rapid laboratory testing procedures. PMID- 16260005 TI - Characterisation of recombinant rat TRPM2 and a TRPM2-like conductance in cultured rat striatal neurones. AB - TRPM2, a member of the TRP ion channel family, is expressed both in the brain and immune cells of the monocyte lineage. Functionally, it is unique in its activation by intracellular ADP-ribose and both oxidative and nitrosative stress. To date studies of this channel have concentrated on human recombinant channels and rodent native preparations. This provides the potential for cross-species complications in the interpretation of native tissue observations based on recombinant channel phenotype. Consequently, we have cloned and heterologously expressed rat TRPM2 (rTRPM2) in HEK293 cells. We find that, like hTRPM2, it responds to intracellular ADP-ribose in a manner dependent on extracellular Ca(2+). At the single channel level rTRPM2 is a slow gating, large conductance (84pS) channel that rapidly runs down in isolated membrane patches. Pharmacologically, rTRPM2 is rapidly and irreversibly blocked by clotrimazole (10muM), thus resembling hTRPM2 but not the TRPM2-like current of the rat-derived insulinoma CRI-G1, which exhibits reversible inhibition by this agent. We show that cultured rat striatal neurones exhibit an ADP-ribose-activated conductance at both the whole cell and single channel level. Pharmacologically this neuronal current can be irreversibly inhibited by clotrimazole. It is also sensitive to removal of extracellular Ca(2+), suggesting that it is mediated by TRPM2 containing channels. These data provide a functional characterisation of heterologously expressed rTRPM2 and demonstrate that, in addition to the previous descriptions in immune cells, microglia and insulinomas, a TRPM2-like conductance can be found in neurones derived from the rodent CNS. PMID- 16260006 TI - Parietal rTMS distorts the mental number line: simulating 'spatial' neglect in healthy subjects. AB - Patients with left-sided visuospatial neglect, typically after damage to the right parietal lobe, show a systematic bias towards larger numbers when asked to bisect a numerical interval. This has been taken as further evidence for a spatial representation of numbers, perhaps akin to a mental number line with smaller numbers represented to the left and larger numbers to the right. Previously, contralateral neglect-like symptoms in physical line bisection have been induced in healthy subjects with repetitive transcranial magnetic stimulation (rTMS) over right posterior parietal lobe. Here we used rTMS over parietal and occipital sites in healthy subjects to investigate spatial representations in a number bisection task. Subjects were asked to name the midpoint of numerical intervals without calculating. On control trials subjects' behaviour was similar to performance reported in physical line bisection experiments. Subjects underestimated the midpoint of the numerical interval. Repetitive transcranial magnetic stimulation produced representational neglect like symptoms in number bisection when applied over right posterior parietal cortex (right PPC). Repetitive TMS over right PPC shifted the perceived midpoint of the numerical interval significantly to the right while occipital TMS had no effect on bisection performance. Our study therefore provides further evidence that subjects use spatial representations, perhaps akin to a mental number line, in basic numerical processing tasks. Furthermore, we showed that the right posterior parietal cortex is crucially involved in spatial representation of numbers. PMID- 16260007 TI - Signals that influence food intake and body weight. AB - Energy homeostasis is a complex on-going process that includes maintaining immediately available as well as stored nutrient levels at optimal levels given the environment. To accomplish this, the brain receives continuous information about stored energy and current and anticipated fluxes in critical organs, as well as about food that is potentially available or being eaten and absorbed. The brain in turn determines when and how much food will be consumed, balancing this activity with other behaviors. This review discusses hormonal and related satiety signals generated as food is being consumed, and upon adiposity signals related to the amount of fat stored in the body, that influence energy intake and ultimately body weight. PMID- 16260008 TI - Early influences on human energy regulation: thrifty genotypes and thrifty phenotypes. AB - Early influences on human ingestive behavior and other aspects of energy homeostasis can be defined according to two very different time scales: the evolutionary time frame responsible for selection of behavioral and metabolic traits embedded within the genome; and the life-course time frame responsible for setting the phenotype. Evolutionary influences: Famine has been a constant threat to human survival leading to the selection of thrifty genes. Thriftiness can take many forms: metabolic (an 'energy-sparing' super-efficient metabolism); adipogenic (a propensity to rapid fat gain); physiologic (an ability to switch off non-essential processes); gluttony (a tendency to gorge when food is available); sloth (a tendency to conserve energy through inactivity); or behavioral (hoarding, meanness, theft, etc). Life-course influences: The nutritional environment of the early embryo can have a major impact on its survival, and its immediate and later physiology. Subsequently, the fetus is sensitive to its nutrient supply that in turn is affected by maternal fuel supply and by the constraints of the utero-placental unit. Adaptive plasticity also continues through infancy. Ingestive behavior in terms of appetite and satiety could theoretically be affected by some of these metabolic adaptations. This paper will describe the key elements of the thrifty genotype and phenotype and review the evidence base relating these early effects to differences in ingestive behavior. PMID- 16260009 TI - Vaccination coverage in the South-East Anatolian Project (SEAP) region and factors influencing low coverage. AB - OBJECTIVES: To determine the vaccination coverage of children living in the South east Anatolian Project (SEAP) region; whether the vaccination coverage was similar to formal reports, other studies and other countries; and which factors influence vaccination, in order to indicate how vaccination coverage can be improved. STUDY DESIGN: A descriptive cross-sectional study conducted in nine provinces of the SEAP region in order to determine public health problems and their causes. METHODS: A population-based sample of 1150 houses was selected from rural and urban areas of the SEAP region and visited by the researchers. Questionnaires were applied in 2001 and 2002. RESULTS: In the SEAP region, only 30% of children had received a complete set of vaccines. The vaccination coverage was 76.7% for Bacille Calmette-Guerin; 62.0% for the third doses of diphtheria, tetanus toxoid, pertussis and polio vaccine; 62.7% for measles; 44% for the third dose of hepatitis B vaccine in children aged 12-23 months; and 13.3% for the second dose of tetanus toxoid in women who gave birth in the last 5 years. In logistic regression analysis, residence type, number of siblings, birth interval, follow-up visits of midwives, and maternal level of education were found to influence whether children were completely vaccinated. CONCLUSIONS: The findings of this study indicate that vaccination coverage is not acceptable in the SEAP region. Efforts must focus on family planning services, education of women, follow-up visits and strengthening health facilities, especially in rural regions, to improve vaccination. PMID- 16260011 TI - Rationales for home safety promotion in the Iranian primary healthcare system: results from a pilot study. AB - OBJECTIVE: To analyse the prerequisites for a nationwide primary healthcare (PHC) home safety promotion programme in Iran. BACKGROUND: Injury is a major public health problem throughout the world, currently accounting for one-seventh of all premature deaths and disabilities. Within 20 years, it is estimated that the proportion will increase to one-fifth. The present healthcare system in Iran was started in 1979, with a major focus on easy access to services and prevention. The system is based on the 'health house', which is run by community health workers. A survey shows that 36% of injuries occur in the home environment. A pilot phase of the Home Safety Promotion Programme was initiated in 1994, and included safety checking at home for fences, kitchens, drugs and poisons, heaters, electricity, and stairs and ladders. The pilot study covered 478,551 households out of the 12 million (approximately) in Iran. Sixty-nine supervisors were involved individually, assembled into eight focus groups. RESULTS: Household safety increased by 10-20% over the 4 years of the study. The frequency of home visits changed from annual to seasonal, since all participants agreed that there were seasonal differences in safety problems. The supervisors showed a high level of knowledge of injury as a public health problem, and also positive attitudes towards doing something about safety on the basis of a PHC scheme. The role of a surveillance system was highlighted, and it was suggested that such a system should be added to the programme. DISCUSSION AND CONCLUSION: Based on our preliminary findings, there were reasons to obtain a policy decision concerning a national programme for safety promotion before extending the pilot scheme to the whole country. A national safety programme was decided upon following completion of the pilot study. It includes a home-related-injury surveillance system that is mandatory in rural areas and voluntary in some cities. PMID- 16260010 TI - The epidemiology of moderate and severe injuries in a Nicaraguan community: a household-based survey. AB - Although injuries constitute a major public health problem worldwide, the magnitude and nature of this problem is difficult to establish in low-income settings due to the lack of valid and representative data. In Nicaragua, several parallel studies have been carried out attempting to assess levels and patterns of injury using various sources of information. The objective of this study was to describe the magnitude and profile of moderate and severe injuries in a well defined community in Nicaragua. The study was conducted using a household-based survey design. Randomized cluster sampling provided information from 10,797 households (63,886 inhabitants). The outcomes included fatal and non-fatal injuries registered during a recall period of 6 weeks. Seven percent of all injuries were classified as moderate or severe. The estimated annual incidence rate of moderate/severe injuries was 27.6 per 1000 [95% confidence intervals (CI): 26.4-29.2], while the mortality and impairment rates were 108.9 (95% CI: 83.5-134.4) and 95.3 (95% CI: 71.4-119.2) per 100,000 inhabitants, respectively. Home and traffic areas were the main environments associated with injury occurrence. The most affected groups were the elderly, children and males. No differences were found between urban and rural areas. Only 9% of all cases, including minor injuries, sought hospital treatment. The main causes of non-fatal injuries were falls, traffic and cuts, whereas fatalities were largely associated with intentional injuries. For every death due to injury, there was one permanent disability, 25 moderate/severe injuries and 354 minor injuries. This study provides a broad description of injury magnitudes and patterns in a defined Nicaraguan community, and demonstrates the aggregate injury pyramid of the same community. It also identifies the issue of severity and concludes with a recommendation to apply different criteria of severity. Our results support the call for a careful evaluation of injury data sources and severity scores when planning injury prevention programmes. PMID- 16260013 TI - Treatment of fascioliasis in human infections. PMID- 16260012 TI - The causes and consequences of injury in students at UK institutes of higher education. AB - BACKGROUND: Injuries are the leading cause of mortality and also an important cause of permanent disability in young people living in developed countries. This study aimed to inform injury prevention programmes by assessing the incidence of injury and disabling injury, and identifying factors predicting injury in students at institutes of higher education, an under-researched group with regard to injury studies. METHODS: Secondary analysis of data obtained from a postal questionnaire survey of 1208 students at three institutions in the UK. RESULTS: Eighteen percent (222/1208) of students reported at least one injury requiring medical attention in the last year. Males and younger students were at greater risk, as were those who reported 'seeking out risky activities because they enjoyed potential danger'. Seventy-nine percent of reported injuries caused some degree of, at least temporary, disability. Four percent of students reported an injury in the last year that interfered with studies, work or other regular daily activities for at least 1 month. Team sports increased the risk of injury by a factor of 2.5 independently of age, gender and risk-taking attitudes. Sport/physical activity injuries were much more likely to be disabling than injuries attributable to other causes. CONCLUSIONS: There is a need for injury prevention programmes among students at institutes of higher education. Programmes need to disseminate the risk of disabling injury, particularly that attributable to participation in sport/physical activity. There is an urgent need for those working on injury prevention to collaborate with those working on other aspects of health promotion to define the net health gain from participation in sport/physical activity and to develop coherent public health messages. PMID- 16260014 TI - Helicobacter pylori infection in children: prevalence, diagnosis and treatment outcome. AB - The clinical significance of Helicobacter pylori infection in children remains largely unknown. The rate of acquisition at different ages has not been ascertained using reliable tests on gastric biopsies. We determined prospectively the prevalence of H. pylori infection in children and its association with gastroduodenal disease. We evaluated 240 children undergoing upper gastrointestinal endoscopy for H. pylori infection by rapid urease test, culture, ureA PCR and histopathology. Group I constituted 58 children with upper abdominal pain (UAP) and group II (controls) of 182 children without UAP who underwent diagnostic or therapeutic endoscopy for other reasons. Helicobacter pylori positive children with UAP received anti-H. pylori therapy. Helicobacter pylori infection was significantly higher in children with UAP than controls (53.4% vs. 28%; P<0.001) and overall prevalence increased with age. On follow-up endoscopy, H. pylori had been eradicated from 82% of children with UAP; it was eradicated from the remaining 18% after a second regimen. Treated H. pylori-positive children with UAP remained symptom-free for a median of 25 months. Control children remained chronically H. pylori infected. Chronic inflammation was present in all infected children, and active inflammation in 48.8%. The study shows H. pylori infection increases with age and is strongly linked to UAP in children. PMID- 16260015 TI - Seroprevalence of Toxoplasma gondii infection among pre-schoolchildren aged 1-5 years in the Democratic Republic of Sao Tome and Principe, Western Africa. AB - The prevalence status of Toxoplasma gondii infection in children of the Democratic Republic of Sao Tome and Principe (DRSTP), Western Africa, is unknown to date. A serologic survey of T. gondii infection among pre-schoolchildren aged <5 years in the DRSTP was assessed by the latex agglutination (LA) test from November 2003 to March 2004. The overall seroprevalence of T. gondii infection was not low, reaching 21.49% (26/121). No significant gender difference in seroprevalence was found between boys (19.30%; 11/57) and girls (23.44%; 15/64) (chi2 = 0.31, P = 0.58). The older age group of 4-5 years had significantly higher seroprevalence (36.67%; 11/30) than the younger age group of <2 years (10.34%; 3/29) (chi2 = 5.64, P = 0.02). It was noteworthy that the majority of seropositive boys (90.91%; 10/11) or older children aged > or = 2 years (82.61%; 19/23) had high LA titres of > or = 1:1024, indirectly indicating acute Toxoplasma infection. This study is the first report indicating that T. gondii infection is not low in pre-schoolchildren aged <5 years in the DRSTP. Whether the DRSTP pre-schoolchildren acquire T. gondii infection through constant exposure to the parasite from their daily activities owing to poor environmental hygiene should be further evaluated. PMID- 16260017 TI - Light and electron microscopy studies of the oesophagus and crop epithelium in Aplysia depilans (Mollusca, Opisthobranchia). AB - The oesophagus and crop epithelium of Aplysia depilans consist in a single layer of columnar cells with apical microvilli, and some of them also possess cilia. Cell membrane invaginations, small vesicles, multivesicular bodies and many dense lysosomes were observed in the apical region of the cytoplasm. In most cells, a very large lipid droplet was observed above the nucleus and a smaller one was frequently found below the nucleus; glycogen granules are also present. Considering these ultrastructural features, it seems that these cells collect nutritive substances from the lumen by endocytosis, digest them in the apical lysosomes and store the resulting products. The cell bodies of mucus secreting flask-shaped cells are subepithelial in the oesophagus and intraepithelial in the crop. Histochemistry methods showed that the secretion stored in these cells contains acidic polysaccharides. Secretory vesicles with thin electron-dense filaments scattered in an electron-lucent background fill most of these cells, and the basal nucleus is surrounded by dilated rough endoplasmic reticulum cisternae containing small tubular structures. Considering the relatively low number of secretory cells, mucus production cannot be high. Moreover, since protein secreting cells were not observed in either oesophagus or crop, extracellular digestion in the lumen of these anterior segments of the digestive tract most probably depend on the enzymes secreted by the salivary and digestive glands. PMID- 16260016 TI - Comment on: Strongyloidiasis: a mistaken diagnosis and a fatal outcome in a patient with diarrhoea. PMID- 16260018 TI - Interactive toxicity of chlorpyrifos and parathion in neonatal rats: role of esterases in exposure sequence-dependent toxicity. AB - We previously reported that sequence of exposure to chlorpyrifos and parathion in adult rats can markedly influence toxic outcome. In the present study, we evaluated the interactive toxicity of chlorpyrifos (8 mg/kg, po) and parathion (0.5 mg/kg, po) in neonatal (7 days old) rats. Rats were exposed to the insecticides either concurrently or sequentially (separated by 4 h) and sacrificed at 4, 8, and 24 h after the first exposure for biochemical measurements (cholinesterase activity in brain, plasma, and diaphragm and carboxylesterase activity in plasma and liver). The concurrently-exposed group showed more cumulative lethality (15/24) than either of the sequential dosing groups. With sequential dosing, rats treated initially with chlorpyrifos prior to parathion (C/P) exhibited higher lethality (7/23) compared to those treated with parathion before chlorpyrifos (P/C; 1/24). At 8 h after initial dosing, brain cholinesterase inhibition was significantly greater in the C/P group (59%) compared to the P/C group (28%). Diaphragm and plasma cholinesterase activity also followed a relatively similar pattern of inhibition. Carboxylesterase inhibition in plasma and liver was relatively similar among the treatment groups across time-points. Similar sequence-dependent differences in brain cholinesterase inhibition were also noted with lower binary exposures to chlorpyrifos (2 mg/kg) and parathion (0.35 mg/kg). In vitro and ex vivo studies compared relative oxon detoxification of carboxylesterases (calcium-insensitive) and A-esterases (calcium-sensitive) in liver homogenates from untreated and insecticide pretreated rats. Using tissues from untreated rats, carboxylesterases detoxified both chlorpyrifos oxon and paraoxon, while A-esterases only detoxified chlorpyrifos oxon. With parathion pretreatment, A-esterases still detoxified chlorpyrifos oxon while liver from chlorpyrifos pretreated rats had little apparent effect on paraoxon. We conclude that while neonatal rats are less capable than adults at detoxifying many organophosphorus insecticides including chlorpyrifos and parathion, toxicant-selective differences in detoxification play a role in sequence-dependent toxicity in both neonatal and adult rats with these two insecticides. PMID- 16260019 TI - Reconstitution of active diphtheria toxin based on a hexahistidine tagged version of the B-fragment produced to high yields in bacteria. AB - Diphtheria toxin consists of an A-fragment that inactivates elongation factor 2 and a B-fragment that both binds to the toxin receptor and mediates translocation of the A-fragment across cellular membranes to the cytosol. Several fragments of the toxin and an inactive version of the holotoxin have been expressed in Escherichia coli, but the B-fragment alone has proven difficult to express. Only low levels of expression have been achieved. We have designed a hexahistidine tagged version of a modified diphtheria toxin B-fragment (DT-BHis) that can be expressed to high levels in E. coli. DT-BHis contains the entire diphtheria toxin B-fragment preceded by an alanine and succeeded by a leucine, a glutamic acid and a hexahistidine tag and could be purified in a single step using nickel-coated agarose beads to 85% homogeneity. DT-BHis bound specifically to the diphtheria toxin receptor and was able to compete out the effect of the wild type diphtheria toxin. Furthermore, DT-BHis was able to form pores in cellular membranes in a manner similar to the wild type B-fragment. The high yield makes DT-BHis a suitable tool in studies of diphtheria toxin interaction with cells or liposomes since functional diphtheria toxin was easily formed upon addition of A-fragment. The reconstituted diphtheria toxin showed toxicity in the same range as the wild type. PMID- 16260020 TI - Effect of pepsin digestion on the antivenom activity of equine immunoglobulins. AB - Enzyme digestion of animal-derived sera followed by antibody purification is a classical process used to prepare snake antivenoms worldwide. In this work, we have studied the effect of the harsh conditions prevailing during the digestion step on the activity of the final product, F(ab')(2). To this purpose, the recovery of the activity of anti-Bothrops hyperimmune equine plasma was determined after pepsin digestion under different sets of processing conditions. The balance between pH level and reaction time was found to be critical, reflecting a compromise between complete cleavage of immunoglobulins and strong denaturation of the F(ab')(2) fragments. For pH in the range 2.8-3.2, 30-65% of the initial activity was lost depending mainly on the processing time, as determined by a competition ELISA technique. Pepsin digestion was also carried out with purified immunoglobulins from the same plasma. SDS PAGE run on the digested immunoglobulins allowed us to verify that the lightest isotypes were more resistant to digestion than the heavier ones. In conclusion, for equine F(ab')(2) antivenom production, it seems convenient to carry out digestion at pH values sufficiently low to ensure that total IgG breakdown is achieved in the shortest time compatible with precise operation in the production scale. PMID- 16260021 TI - HIV-1 gp120 induces NFAT nuclear translocation in resting CD4+ T-cells. AB - The replication of human immunodeficiency virus (HIV) in CD4+ T-cells is strongly dependent upon the state of activation of infected cells. Infection of sub optimally activated cells is believed to play a critical role in both the transmission of virus and the persistence of CD4+ T-cell reservoirs. There is accumulating evidence that HIV can modulate signal-transduction pathways in a manner that may facilitate replication in such cells. We previously demonstrated that HIV gp120 induces virus replication in resting CD4+ T cells isolated from HIV-infected individuals. Here, we show that in resting CD4+ T-cells, gp120 activates NFATs and induces their translocation into the nucleus. The HIV LTR encodes NFAT recognition sites, and NFATs may play a critical role in promoting viral replication in sub-optimally activated cells. These observations provide insight into a potential mechanism by which HIV is able to establish infection in resting cells, which may have implications for both transmission of HIV and the persistence of viral reservoirs. PMID- 16260022 TI - Quantification of stato-kinetic dissociation by semi-automated perimetry. AB - The difference in threshold sensitivities that are found when examining the visual field (VF) with static versus kinetic perimetric methods is called stato kinetic dissociation (SKD). In this pilot study, we describe a semi-automated procedure for quantifying SKD. Fifteen patients with VF defects were examined with kinetic and static perimetry. SKD values were defined as positive when the static scotoma was larger than the kinetic one. We found significant local variations of SKD along scotoma borders with the individual reaction time as an important criterion when determining kinetic thresholds. There was a verifiable SKD in all patients with locally negative values in eight subjects. PMID- 16260023 TI - Impaired visual processing of letter and digit strings in adult dyslexic readers. AB - The slow word reading of developmental dyslexics may stem from a string processing impairment which in turn reflects visual attentional deficits. We indeed found substantially enhanced recognition time thresholds in the dyslexic adult readers. However, their position profiles were hard to reconcile with any of the discussed visual attentional deficit hypotheses and with the prediction that dyslexic readers suffer from an absent string processing system as they exhibited similar M-shaped position profiles for digit and letter strings as the normal reading controls. PMID- 16260024 TI - Effects of local and global factors in the Pinna illusion. AB - The Pinna illusion (Pinna & Brelstaff, 2000) consists of two concentric rings of micropatterns that appear to counter-rotate when the observer moves towards the stimulus. There have been several reports that the illusion is stronger when the retinal expansion is produced by observer self-motion than when produced on a computer screen without observer self-motion. In fact, we found that the illusion is as strong (or stronger) when the retinal expansion is produced on a computer screen without observer self-motion. In a second series of experiments the strength of the Pinna illusion was inferred from the amount of physical counter rotation required to null it. The strength of the illusion is relatively unaffected by changes to the global structure of the display but minor changes to the micropatterns comprised in the display can effectively eliminate the illusion. We provide a simple model of optical flow that is in very good agreement with many of the results reported. PMID- 16260025 TI - Antisaccade velocity, but not latency, results from a lack of saccade visual guidance. AB - Antisaccades are slower in peak velocity, more dysmetric, and longer in latency than prosaccades. This study used a novel visually guided antisaccade task to determine how visual target presence affects antisaccade metrics. The results showed that peak velocity and endpoint error of visually guided antisaccades were more similar to prosaccades than to traditional antisaccades, whereas their latencies were similar to those of traditional antisaccades. The velocity of prosaccades, and to a lesser extent that of antisaccades, were boosted by the sudden appearance of a target. These results suggest that the lower velocity and increased dysmetria of traditional antisaccades result from the absence of a visual target, but their longer latency is more likely a result of suppressing a prosaccadic reflex. PMID- 16260026 TI - Yellow pigments used in rapid identification of aflatoxin-producing Aspergillus strains are anthraquinones associated with the aflatoxin biosynthetic pathway. AB - Studies on biological control of aflatoxin production in crops by pre-infection with non-toxigenic Aspergillus flavus strains have created a need for improved methods to screen isolates for aflatoxigenicity. We have evaluated two empirical aflatoxigenicity tests: (i) yellow pigment production, and (ii) the appearance of a plum-red color in colonies exposed to ammonium hydroxide vapor. Yellow pigments from aflatoxigenic A. flavus were shown to function as pH indicator dyes. Seven pigments representing most of the pigmentation in extracts have been isolated using color changes when chromatography spots were exposed to ammonium hydroxide vapor to guide fractionation. Their structures have been shown to be norsolorinic acid, averantin, averufin, versicolorin C, versicolorin A, versicolorin A hemiacetal and nidurufin, all of which are known anthraquinone pigments on, or associated with, the aflatoxin biosynthetic pathway in Aspergillus spp. Thus, the basis of both empirical tests for aflatoxigenicity is detecting production of excess aflatoxin biosynthetic intermediates. PMID- 16260027 TI - Hydrazide derivatives produce active oxygen species as hydrazine. AB - It is well documented that some hydrazines are quite sensitive to oxidation and may serve as the electron donor for the reduction of oxygen, whereas hydrazides are not believed to react directly with oxygen. Data presented in this paper show that both hydrazides and hydrazines share an N-N moiety, which is assumed to react with atmospheric oxygen and produce oxygen radicals, at various degrees of efficiency. Since spectrometric measurements of hydrazide just after solubilization showed that the molecular mass remains constant in the absence of oxygen, we can conclude that hydrazides do not react with the oxygen through a slow spontaneous hydrolytic release of hydrazine. However, hydrazine is more reactive than hydrazide, which requires hours rather than minutes to produce measurable quantities of radical species. Differences were also apparent for various substituted derivatives. The reaction was significantly enhanced by the presence of metal ions. Data reported here demonstrate that hydrazides cause irreversible damage to the prosthetic group of proteins as well as causing degradation of the polypeptide chain into small fragments. PMID- 16260028 TI - Are pharmaceuticals potent environmental pollutants? Part II: environmental risk assessments of selected pharmaceutical excipients. AB - As part of achieving national environmental goals, the Swedish Government commissioned a report from the Swedish Medical Products Agency on environmental effects of pharmaceuticals and cosmetics and hygiene products. Five excipients used in pharmaceutical products were selected for environmental risk assessments, applying the computer-based model EUSES. Docusate sodium was identified as a possible risk for sediment-dwelling organisms when taking the total amount used into account. Although, experimental toxicity data on sediment-dwelling organism and data on concentrations in sediments are still required before firm conclusions regarding the environmental risk can be made. The environmental risks posed by excipients used in pharmaceutical products are likely to be negligible. This study identifies that knowledge gaps regarding environmental risks posed by pharmaceutical excipients are evident. PMID- 16260029 TI - A phase II study of irofulven in women with recurrent and heavily pretreated ovarian cancer. AB - OBJECTIVE: To determine the safety and efficacy of a novel illudin S derivative, irofulven (MGI-114), in patients with recurrent ovarian cancer who had received extensive prior chemotherapy. METHODS: The trial was an open label phase II study. Patients initially enrolled in this study were treated every 14 days with a dose of 24 mg/m2. Unexpected retinal toxicity associated with this dose and schedule lead to modification of the dosing to 0.55 mg/kg on the same schedule with a maximum individual dose of 50 mg. Dose reductions were permitted based on both hematologic and non-hematologic toxicities. RESULTS: Seventy-four women were accrued and stratified into two cohorts including 58 women with platinum resistant disease and 16 with platinum-sensitive disease. Non-hematologic toxicities included nausea, vomiting, and fatigue. Thirty-one women had between one and six visual symptoms, most were Grade 1 and 2 in nature. The majority of visual toxicities resolved either during treatment or post-treatment with irofulven. There was one partial response in each cohort with 19 (33%) and 8 (50%) of women having stable disease in the platinum-resistant and platinum sensitive cohorts, respectively. CONCLUSIONS: Irofulven at 24 mg/m2 on every 14 day schedule is associated with significant retinal toxicity in this patient population. Dosing at 0.55 mg/kg has persistent retinal toxicity, yet demonstrated only limited anti-tumor activity in a population of women who had received extensive prior chemotherapy. PMID- 16260030 TI - Interest of adolescents in genetic testing for nicotine addiction susceptibility. AB - OBJECTIVES: Genetic tests for nicotine addiction susceptibility may someday offer preventive medicine specialists new tools to reduce smoking among adolescents. This paper examines adolescents' interest in, and reasons behind interest in, such testing and correlates of interest. METHODS: The sample included 211 healthy patients (ages 13-21) recruited from Georgetown University Medical Center's adolescent medicine clinic. Subjects completed a one-time behavioral survey immediately prior to or following a general medical check-up during calendar years 2001-2005. A 4-point self-report survey item served as the dependent variable. RESULTS: Sixty-two percent of adolescents were interested in genetic testing. Among the 72% of adolescents who provided a reason for their interest, 35% would find the information interesting for general or nonspecific reasons, 30% would find it personally useful, 8% noted it would be irrelevant, and 13% stated it would be unimportant; school performance and interest in cancer susceptibility testing were associated with interest in nicotine addiction susceptibility testing (adjusted r2 = 21%; P < 0.0001). CONCLUSIONS: Adolescent primary care patients will likely be receptive to comprehensive tobacco control programs incorporating genetic testing. Higher levels of educational achievement and greater interest in DNA-based preventive medicine may characterize those most interested. Offering testing will be contingent upon the development of safe and effective genetic tests. PMID- 16260032 TI - Injectable and self-curing composites of acrylic/bioactive glass and drug systems. A histomorphometric analysis of the behaviour in rabbits. AB - Injectable self-curing systems based on PMMA, phosphate-free bioactive glasses and the drug fosfosal, a phosphate derivative of salicylic acid with analgesic and moderate anti-inflammatory properties, have been tested in vivo to evaluate their biocompatibility. The model consisted of the injection of dough of cement into a defect created in the femur of rabbits, and the cure of the cement in situ after implantation. The biological response was studied in the short and long terms by macroscopic, radiological and histopathological examination, and quantitatively by histomorphometric and statistical analysis considering the most representative variables at the bone-cement interface: cement, bone marrow, newly formed bone and connective tissue. All bioactive formulations presented resorption of the cement at the end of the experiment in contrast to the control of PMMA, due to the presence of resorbable components. The presence or absence of the phosphate group added by the drug fosfosal influenced mainly on the new bone formation process. The cement formulated with bioactive glasses and in absence of fosfosal produced the maximum amount of neoformed bone at 2 weeks, and then it resorbed at 4 weeks to give a higher amount of neoformed bone at the end of the experiment, compared with the formulation containing only fosfosal. The presence of fosfosal and bioactive glass together affected the ossification process strongly. The osseous tissue was produced more gradually but it continuously increased giving rise to a more stable bone at the end of the experiment. PMID- 16260033 TI - Fatigue and life prediction for cobalt-chromium stents: A fracture mechanics analysis. AB - To design against premature mechanical failure, most implant devices such as coronary and endovascular stents are assessed on the basis of survival, i.e., if a fatigue life of 10(8) cycles is required, testing is performed to ascertain whether the device will survive 10(8) cycles under accelerated in vitro loading conditions. This is a far from satisfactory approach as the safety factors, which essentially tell you how close you are to failure, remain unknown; rather, the probability of fatigue failure should instead be assessed on the basis of testing to failure. In this work, a new damage-tolerant analysis of a cardiovascular stent is presented, where the design life is conservatively evaluated using a fracture mechanics methodology. In addition to enabling estimates of safe in vivo lifetimes to be made, this approach serves to quantify the effect of flaws in terms of their potential effect on device failure, and as such provides a rational basis for quality control. PMID- 16260034 TI - RNA interference targeting of Bcr-Abl increases chronic myeloid leukemia cell killing by 17-allylamino-17-demethoxygeldanamycin. AB - 17-Allylamino-17-demethoxygeldanamycin (17-AAG) induces degradation of Hsp90 client proteins, including Bcr-Abl, however, its clinical use as an anti-tumor agent may be limited by toxicity and modest efficacy. We reasoned that Bcr-Abl targeting by RNA interference (RNAi) might selectively increase the activity of 17-AAG against Bcr-Abl+ leukemia cells. 17-AAG in combination with targeting small interfering RNAs (siRNAs) reduced Bcr-Abl protein levels, triggered increases in markers of apoptosis and decreased cell viability more effectively than did control siRNA and 17-AAG together, or Bcr-Abl targeting siRNA alone. Combination targeting strategies such as this may therefore achieve enhanced therapeutic potency. PMID- 16260035 TI - Depsipeptide-resistant KU812 cells show reversible P-glycoprotein expression, hyper-acetylated histones, and modulated gene expression profile. AB - Depsipeptide (FK228), a histone deacetylase inhibitor, is a promising new anticancer agent. The mechanism of resistance to this agent was studied using KU812 cells. Depsipeptide-resistant KU812 cells expressed P-glycoprotein (P-gp) and their resistance was abolished by co-treatment with verapamil. P-gp expression returned to the parental cell level when resistant cells were cultured in depsipeptide-free medium, while resistant cells cultured in the medium containing 16 nM depsipeptide still showed hyper-acetylation of histones. Moreover, resistant cells showed erythroid differentiation. Microarray analysis revealed that 28 genes showed increased expression and three genes showed decreased expression in resistant cells compared with parental cells. These 31 genes had various functions relating to signal transduction, cell cycle, apoptosis, and control of cell morphology and differentiation. Among the 28 genes that were upregulated, 15 genes also showed an increased expression in parental cells treated with 4 nM depsipeptide for 48 h, while the other 13 genes including P-gp were different. Among the three genes with decreased expression, HEP27 was most dramatically downregulated. These findings suggest that continuous exposure to depsipeptide reversibly induces P-gp, which contributes to the onset of resistance, but the altered gene expression profile of resistant cells may also play a role. PMID- 16260036 TI - Trends in children's disclosure of abuse in Israel: a national study. AB - OBJECTIVE: To identify characteristics of suspected child abuse victims that are associated with disclosure and nondisclosure during formal investigations. METHODOLOGY: The database included all suspected cases of physical and sexual abuse investigated in the state of Israel between 1998 and 2002. All investigative interviews were conducted using a single standardized protocol, the National Institute of Child Health and Human Development (NICHD) Investigative Interview Protocol. FINDINGS: Overall, 65% of the 26,446 children made allegations when interviewed, but rates of disclosure were greater in the case of sexual (71%) than physical (61%) abuse. Children of all ages were less likely to disclose/allege abuse when a parent was the suspected perpetrator. Rates of disclosure/allegation increased as children grew older, with 50% of the 3- to 6 year-olds, 67% of the 7- to 10-year-olds, and 74% of the 11- to 14-year-olds disclosing abuse when questioned. CONCLUSIONS: Although most interviews of suspected victims yielded allegations, such rates of disclosure varied systematically depending on the nature of the alleged offences, the relationship between alleged victims and suspected perpetrators, and the age of the suspected victims. The findings obtained in this large and unselected data set confirm patterns previously reported in smaller and quite selective samples, most of them obtained in the United States. PMID- 16260037 TI - The future of the protocol movement: commentary on Hershkowitz, Horowitz, and Lamb (2005). PMID- 16260038 TI - The effects of early sexual abuse on adult risky sexual behaviors among persons with severe mental illness. AB - OBJECTIVE: There were two aims: first, to examine the relationship between prior sexual abuse and three types of adult risky sexual behaviors [(1) ever traded sex for drugs or money, (2) had unprotected sex in the past 6 months, and (3) frequency of unprotected sex in the past 6 months] among persons with severe mental illness (SMI), and second, to examine the potential mediating effects of adult rape, substance use, and PTSD. METHOD: Using a pooled sample of individuals with SMI (N=609), logistic and negative binomial regression analyses were used to investigate the impact of prior sexual abuse on these adult risky sexual behaviors. RESULTS: Childhood sexual abuse was associated with having ever traded sex for money and having engaged in unprotected sex in the past 6 months. However, childhood sexual abuse was inversely associated with the number of times males had unprotected sex in the past 6 months. Results differed between males and females and the impact of potential mediators also varied by gender and type of outcome studied. CONCLUSION: These findings suggest a complex link between childhood sexual abuse and adult risky sexual behaviors in persons with SMI. Clinical assessments of child abuse sequelae should include a variety of indicators and parameters of adult risky sexual behavior, as persons with SMI are at an increased risk of engaging in high-risk sexual behaviors and tend to have a higher exposure to childhood sexual abuse than does the general population. PMID- 16260039 TI - Childhood sexual abuse, social support, and psychological outcomes: a loss framework. AB - OBJECTIVE: The objectives of the study were: (a) to develop a scale to assess CSA related loss among a college sample of CSA survivors (CSALM), (b) to examine the measure's convergent validity through associations among depression, alexithymia, coping, and social support, and (c) to test whether social support moderates the relation between multiple experiences of CSA and loss. METHOD: The study involved a survey methodology and included college-age women (n=116) reporting CSA experiences. RESULTS: Based on the Sexual Victimization Questionnaire (SVQ; [Finkelhor, D. (1979). Sexually victimized children. New York: The Free Press]), 90% of the sample reported CSA before age of 12, 12.3% (n=15) reported CSA before age 12 with an adult over 16, and 42.2% (n=49) reported CSA after age 12 with an adult. Exploratory Factor Analysis of the CSALM revealed a three-factor solution: (a) Loss of Optimism, (b) Loss of Self, and (c) Loss of Childhood. Convergent validity of several scales was evidenced through associations with depression, alexithymia, coping, and social support. Social support from family and friends was found to moderate the association between CSA experiences and loss dimensions. CONCLUSIONS: Findings provide preliminary psychometric support for the CSALM, a tool that will be useful in future investigations of loss among college-age CSA survivors. Findings also support using a loss framework to understand the current mental health of these survivors. PMID- 16260040 TI - Dating violence and sexual risk behaviors in a sample of at-risk Israeli youth. AB - OBJECTIVE: This exploratory study examines the reported dating violence and its association with sexual risk behavior among Israeli adolescents, who are at risk for dropping out of school. METHODOLOGY: A convenience sample of 105 at-risk youth (51 boys and 54 girls) completed self-administered anonymous, questionnaires in small same-gender groups. The questionnaire included a Hebrew version of the Conflict in Adolescent Dating Relationships Inventory, a valid scale developed in the US especially for adolescents, the Hebrew version of the Conflict Tactics Scale and the Self-Efficacy to Refuse Sexual Behavior Scale. RESULTS: We found high rates of perpetration of verbal-emotional abuse for both girls and boys. Rates of reported victimization by physical abuse were higher among boys but that by sexual abuse were higher among girls. We found a moderate inverse association between victimization by dating violence and practicing safe sex among girls. There was a similar but much stronger association for boys who were victims of dating violence. CONCLUSIONS: The high rates of various types of dating violence in Israel, at least among at-risk adolescents, should raise concern and motivate designing and implementing specific prevention interventions. The high reported rates of victimization from dating violence among boys are somewhat surprising and call for special attention and more indepth studies. PMID- 16260041 TI - Prolonged dysphagia due to Listeria-rhombencephalitis with brainstem abscess and acute polyradiculoneuritis. AB - We report a case of previously healthy student with acute rhombencephalitis and brainstem abscess caused by Listeria monocytogenes. The disease begun with uncharacteristic prodromal symptoms of gastrointestinal infection followed by headache and vertigo. After hospital admission the patient rapidly deteriorated, presenting pronounced dysphagia and respiratory failure requiring mechanical ventilation. The diagnosis was established upon clinical symptoms of infection, brainstem involvement, typical MRI findings and positive for L. monocytogenes blood culture. Infection was complicated by acute, demyelinating neuropathy, diagnosed upon clinical symptoms of frail palsy confirmed by ENG. Initially introduced empirical doxycyclin/ceftriaxon treatment was subsequently changed to targeted ampicillin/gentamycin therapy, mechanical ventilation, intravenous human immunoglobulin treatment, tracheostomy and endoscopic gastrostomy. Prolonged dysphagia resolved after rehabilitation. After one year the patient remains well with only slight dysmetria. PMID- 16260042 TI - Rapid identification of 14-3-3-binding proteins by protein microarray analysis. AB - The 14-3-3 protein family consists of acidic 30-kDa proteins composed of seven isoforms in mammalian cells, expressed abundantly in neurons and glial cells of the central nervous system (CNS). The 14-3-3 isoforms form a dimer that acts as a molecular adaptor interacting with key signaling components involved in cell proliferation, transformation, and apoptosis. Until present, more than 300 proteins have been identified as 14-3-3-binding partners, although most of previous studies focused on a limited range of 14-3-3-interacting proteins. Here, we studied a comprehensive profile of 14-3-3-binding proteins by analyzing a high density protein microarray using recombinant human 14-3-3 epsilon protein as a probe. Among 1752 proteins immobilized on the microarray, 20 were identified as 14-3-3 interactors, most of which were previously unreported 14-3-3-binding partners. However, 11 known 14-3-3-binding proteins, including keratin 18 (KRT18) and mitogen-activated protein kinase-activated protein kinase 2 (MAPKAPK2), were not identified as a 14-3-3-binding protein. The specific binding to 14-3-3 of EAP30 subunit of ELL complex (EAP30), dead box polypeptide 54 (DDX54), and src homology three (SH3) and cysteine rich domain (STAC) was verified by immunoprecipitation analysis of the recombinant proteins expressed in HEK293 cells. These results suggest that protein microarray is a powerful tool for rapid and comprehensive profiling of 14-3-3-binding proteins. PMID- 16260043 TI - The use of the GBI as predictor of bipolar disorder in a population of adolescent offspring of parents with a bipolar disorder. AB - OBJECTIVE: To assess the usefulness of the General Behavior Inventory (GBI) to predict the development of mood disorders in the offspring of parents with bipolar disorder. METHOD: The GBI and the K-SADS (first measurement) and the SCID (last measurement) were used to assess psychopathology among 129 adolescent and young adult offspring of a bipolar parent with an interval of 5 years. Based on the SCID results at the last measurement, the offspring were assigned to one of four groups: with bipolar mood disorder, with unipolar mood disorders, with non mood disorders and without disorders and GBI-scores at the first measurement were compared across the four groups. RESULTS: The scores on the Depression scale of the GBI for the offspring who later developed a bipolar or any mood disorder were significantly higher than for the offspring who did not develop a mood disorder across a 5-year interval. For the offspring with a unipolar mood disorder at the first measurement, the scores on the Depression scale were significantly higher for those who switched to bipolar disorder versus those who remained unipolar. CONCLUSIONS: The GBI can be used in a high-risk sample of offspring of parents with bipolar disorder as a self-report measure as an aid to detect those who will develop bipolar disorder across a 5-year interval. PMID- 16260044 TI - Molecular cloning and tissue expression of canine Toll-like receptor 2 (TLR2). AB - Toll-like receptor 2 (TLR2) is involved in an important role on inflammatory reactions against various pathogenic microorganisms. Although some investigations suggested a relation between human and murine TLR2 gene polymorphisms and infections or inflammatory reactions, canine TLR2 gene has not been investigated. In the present study, therefore, the canine full-length of cDNA of Toll-like receptor 2 gene was sequenced and the expression of canine TLR2 mRNA was investigated in various tissues. Canine TLR2 clone was 2552bp encoding 785 amino acids. The similarities of canine TLR2 with human, mouse, rat, cattle, horse and pig TLR2 were 87.6, 72.9, 73.2, 83.1, 85.1 and 80.7% at the nucleotide sequence level, and 84.6, 68.7, 68.5, 78.8, 82.9 and 79.0% at amino acid sequence level, respectively. By a semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) analysis, mRNA of canine TLR2 was detected in blood mononuclear cells (BMCs), lymph node, lung, liver, spleen, bladder, pancreas, small intestine, large intestine and skin of the dogs. PMID- 16260045 TI - Tolerance induction through simultaneous double bone marrow transplantation with two-signal blockade. AB - Cotransplantation of donor bone marrow cells (BMCs) in allograft recipients is currently the most promising concept for clinical tolerance induction; however, it still has many difficulties in its successful performance due to the toxicity of the required host conditioning, the risk of engraftment failure, and the problem of graft-versus-host disease (GVHD), as well as the limited accessibility of donor bone marrow cells. Therefore, we performed the studies to determine whether BMCs from multi-donors are simultaneously engrafted and lead to induction of chimerism-based tolerance through the tolerogenic protocol of whom effectiveness we have shown in a previous study. Using a murine model, it was demonstrated that grafted BMCs from BALB/c (H-2(d)) and CBA mice (H-2(k)) establish mixed type and multi-lineage double chimerism and induce immunological donor-specific tolerance to fully MHC-mismatched skin allografts in host C57BL/6 mice (H-2(b)) receiving conditioning with Busulfan and treatment with the two signal blockade comprised of anti-CD45RB and anti-CD154 monoclonal antibodies. PMID- 16260046 TI - Historical review: Negative efficacy and the constitutive activity of G-protein coupled receptors. AB - The idea that a receptor can produce signalling without agonist intervention and that several antagonists can be 'active' in repressing such spontaneous activity is contained in the concept of ligand-induced conformational changes. Yet, this idea was neglected by pharmacologists for many years. In this article, we review the events that brought inverse agonism and constitutive activity to general attention and made this phenomenon a topic of current research. We also suggest a classification of antagonists based on the cooperativity that links their primary site of interaction with other functional domains of the receptor. PMID- 16260047 TI - A therapeutic role for cannabinoid CB1 receptor antagonists in major depressive disorders. AB - Cannabinoid receptors in the CNS have been implicated in the control of appetite, cognition, mood and drug dependence. Recent findings support the hypothesis that cannabinoid CB1 receptor blockade might be associated with antidepressant and anti-stress effects. A novel potential antidepressant drug class based on this mechanism is supported by the neuroanatomical localization of CB1 receptors and signal transduction pathways that are involved in emotional responses, together with the antidepressant-like neurochemical and behavioral effects induced by CB1 receptor antagonists. Selective CB1 receptor antagonists are in development for the treatment of obesity and tobacco smoking, and could be tested for antidepressant efficacy because recent results of clinical studies suggest that they would also treat comorbid symptoms of depression such as cognitive deficiencies, weight gain, impulsivity and dependence disorders. Thus, CB1 receptor antagonism might constitute an integrated pharmacotherapeutic approach that impacts the affective, cognitive, appetitive and motivational neuronal networks involved in mood disorders. PMID- 16260049 TI - Chronic toxicity of uranium to a tropical green alga (Chlorella sp.) in natural waters and the influence of dissolved organic carbon. AB - The chronic toxicity (72-h cell division rate) of uranium (U) to the unicellular alga, Chlorella sp., was assessed in natural Magela Creek water (NMCW) to provide data for the derivation of a site-specific water quality trigger value for U in Magela Creek, NT, Australia. In addition, the data were compared to those for Chlorella sp. when tested for U toxicity using synthetic Magela Creek water (SMCW), which simulates the inorganic composition of Magela Creek water and contains no organic component. Based on one rangefinder and four definitive toxicity tests, concentrations causing a 50% inhibition of algal growth after 72 h exposure (72 h IC50s) ranged between 137 and 238 microg/LU, no-observed-effect concentrations (NOECs) from 72 to 157 microg/LU and lowest-observed-effect concentrations (LOECs) from 120 to 187 microg/LU. Based on these data, Chlorella sp. was the second most sensitive organism to U of five local species that have been assessed using NMCW. The U toxicity data for Chlorella sp. were incorporated with existing data for the four other species to derive a site-specific guideline value for Magela Creek that is protective of 99% of species of 6 microg/L. The toxicity of U to Chlorella sp. in NMCW was approximately two to four times lower than in SMCW. Based on geochemical speciation modelling, this difference corresponded to a four-fold decrease in the proportion of free uranyl ion (UO2(2+)) in NMCW compared to SMCW, most likely due to the presence of dissolved organic carbon (DOC) in NMCW. Relatively, large variability in U toxicity across the tests conducted in NMCW was found to be inversely related to DOC concentration (r2 = 0.996, n = 4, P = 0.002). Speciation modelling indicated that the increase in DOC was associated with an increase in the proportion of U complexed with DOC (r2 = 0.986, n = 4, P < 0.001) and a decrease in the proportion of the UO2(2+) (r2 = 0.989, n = 4, P = 0.006). When the proportion of UO2(2+) was regressed against U toxicity, a very strong, positive relationship was observed (r2 = 1, n = 4, P < 0.001). The results indicate that the bioavailability and toxicity of U is highly influenced by dissolved organic matter and that the relationship should be further quantified. PMID- 16260050 TI - Study on the constructions and activities of three novel hGHRH analogs with N terminal prolyl modulation+. AB - Growth hormone releasing hormone is one of the hormones secreted by the hypothalamus. Because of its potential applications in agriculture and medicine, its short half-life and its expensive chemical synthesis, an analog with high GHRH activity and prolonged half-life has been looked for. The fusion partner gene with 127 amino acid residues of the C-terminus from L-asparaginase was recombined respectively with asp-pro-pro-hGHRH(1-44), asp-pro-hGHRH(1-44) or asp 1pro-GHRH(2-44) genes synthesized by PCR method to form three kinds of fusion proteins with unique acid labile linker Asp-Pro. The Pro-Pro-hGHRH(1-44), Pro hGHRH(1-44), and 1Pro-GHRH(2-44) peptides were purified to homogeneity by means of cell disruption, washing of inclusion body, ethanol fraction precipitation, acid hydrolysis, SP-Sephadex C-25 and Sephadex G-10 column chromatography. The peptide molecular mass of 5235, 5139 or 4975 Da was determined by ESI mass spectroscopy and purity was determined by SDS-PAGE. In the study of in vitro activity, the antiserum kit against human GH and peptide doses of 0.1, 1.0 and 10 microg/ml were used. These peptides obviously increased GH releases both from human pituitary and from rat pituitary. The activity comparisons showed that there was significant difference between Pro-Pro-hGHRH(1-44)-Gly-Gly-Cys and Pro Pro-hGHRH(1-44) at 1.0 microg/ml, or between 1Pro-hGHRH(2-44) and Pro-Pro-hGHRH(1 44) or Pro-hGHRH(1-44) at 10 microg/ml. The structure-activity relationships showed that at the original C-terminus, for rat pituitary the activity of the GHRH analog with 1Tyr-->Pro was more than that of Pro-Pro-hGHRH(1-44) or Pro hGHRH(1-44). The results showed that the analogs had good GH-releasing activity and species specificity. PMID- 16260051 TI - Tadalafil improves quality of life and exercise tolerance in idiopathic pulmonary arterial hypertension. AB - Pulmonary arterial hypertension has a poor prognosis quoad vitam et valitudinem. Herein, we report on a middle-aged woman affected by idiopathic pulmonary arterial hypertension whose quality of life and exercise tolerance improved remarkably after a six-month course of treatment with the long-acting phosphodiesterase-5 inhibitor tadalafil. PMID- 16260052 TI - Left ventricular hypertrophy in Caucasian master athletes: Differences with hypertension and hypertrophic cardiomyopathy. AB - AIM: To study, by conventional echocardiography, left ventricular remodelling and function in master athletes, hypertension and hypertrophic cardiomyopathy. METHODS: We studied 30 master athletes (MA; soccer players; mean age 43.9+/-5.9), 24 subjects with essential hypertension (HYP; 46.6+/-6), 20 patients with hypertrophic cardiomyopathy (HCM; 42.2+/-9) and 30 normal individuals (CG; 43.4+/ 5). An integrated M-mode/two-dimensional echocardiographic analysis was performed to determine chambers dimensions, relative wall thickness (RWT) and left ventricular mass (LVM), indexed to height in meters raised to the power of 2.7 (LVM/h(2.7)). Cut-off levels for LVM/h(2.7) and RWT were defined to assess 4 different patterns of LV geometric remodelling. In addition, we measured indexes of global systolic performance and indexes of global diastolic function. RESULTS: LV wall thickness and LV end-diastolic dimensions were higher in MA than controls, but significantly lower than other groups. LVH/h(2.7) was increased in 79% of HYP and in 95% of HCM, but was within the normal limits in MA. LV geometry was normal in 22 out of 30 MA (73%), while the remaining (8 athletes, 27%) showed a concentric remodelling. Systolic function (FS and EF) was normal in MA, but was slightly reduced in HYP and increased in HCM. Analysis of diastolic function showed an abnormal relaxation pattern in all HYP and 95% of HCM, but was normal in all MA. The ratio between peak filling rate and stroke volume (PFR/SV), a relatively independent index of diastolic function, was significantly greater in hypertensive patients with normal LV remodelling compared to those without it (4+/-0.39 vs. 4.91+/-0.19; P = 0.0002). CONCLUSION: MA showed lower values of wall thickness, LV dimensions and LV mass compared with HYP and HCM. Despite an abnormal remodelling, all the athletes showed a normal systolic and diastolic function. The differential diagnosis between MA, HYP and HCM is feasible by accurate, comprehensive standard Doppler echocardiography. PMID- 16260053 TI - QT interval prolongation and torsade de pointes associated with indapamide. AB - Direct blockade of the delayed rectifier repolarising potassium current is the major underlying mechanism of drug-induced QT interval prolongation. Indapamide is a well known blocker of the slow component of the delayed rectifier current leading to prolongation of cardiac repolarization. The case of an acquired long QT and torsade de pointes ventricular tachycardia in a woman with systemic lupus erythematosus and hypertension receiving prednisolone and indapamide, respectively, is described in the present report. PMID- 16260055 TI - Radiotherapy for the treatment of primary vaginal cancer. AB - Disease control rates and toxicity were evaluated among 28 primary vaginal cancer patients treated with curative intent using radiotherapy. At 5 years, the majority (60%) of patients were disease-free and local control was 73% when a combination of external beam radiotherapy and brachytherapy were used. Serious late toxicity was seen in 11% of patients. PMID- 16260054 TI - The 'QUAD SHOT'--a phase II study of palliative radiotherapy for incurable head and neck cancer. AB - BACKGROUND AND PURPOSE: The primary objective of this study was to estimate the rate of tumour response to a cyclical hypofractionated palliative radiotherapy regimen (QUAD SHOT) in previously untreated patients with incurable squamous cell carcinoma of the head and neck. Secondary objectives were to prospectively evaluate toxicity, quality of life (QoL) and survival in these patients. PATIENTS AND METHODS: The QUAD SHOT consisted of 14 Gy in four fractions, given twice a day and at least 6h apart, for 2 consecutive days. This regimen was repeated at 4 weekly intervals for a further two courses if there was no tumour progression. The QoL tool used was an abbreviation of the EORTC QLQ-C30. RESULTS: Thirty eligible patients (29 Stage IV, 20 performance status 2-3) had at least one treatment and 16 patients completed all three cycles. Sixteen patients (53%) had an objective response (2CR, 14PR) and a further seven had stable disease. Median overall survival was 5.7 months, median progression free survival was 3.1 months. The treatment was very well tolerated, with improved QoL in 11 of 25 evaluable patients (44%). CONCLUSION: The QUAD SHOT regimen is an effective palliative treatment with minimal toxicity and a good response rate, which impacts positively on patients' QoL. PMID- 16260056 TI - Comparing 3DCRT and inversely optimized IMRT planning for head and neck cancer: equivalence between step-and-shoot and sliding window techniques. AB - BACKGROUND AND PURPOSE: To investigate the feasibility and the advantages of using Intensity-Modulated Radiotherapy (IMRT) for the treatment of head-and-neck cancer. Comparing different methods to deliver IMRT in this clinical setting. MATERIALS AND METHODS: Seven patients (four radical; three post-operative), treated on a 6MV Varian Linac (equipped with an 80 leaves MLC) in accordance with a routine 3DCRT plan, were replanned. Original treatment plans were computed to irradiate a primary Planning Target Volume (PTV1, 54 Gy) and then to perform a boost on a PTV2 (radical: 70.2 Gy; post-operative: 64.8 Gy). IMRT dose plans were inversely-optimized using appropriate constraints with the Helios tool on a Varian Eclipse system. Once the optimal fluences were calculated, different modalities to deliver IMRT were considered: Sliding Window (SW) and Step and Shoot (SS) techniques using a different number of intensity levels to approximate the optimal fluences (e.g. 5, 10 and 20). Mean dose, maximum dose and a number of dose-volume parameters regarding CTV1, CTV2, PTV1, PTV2, OARs (spinal and planning spinal cord, parotids, optical structures, brain and temporal mandibular joint) were considered to compare the five modalities (3DCRT, SW, SS5, SS10, SS20); the Conformity Index (CI), the Irradiated Volume (IV) and the Treated Volume (TV) were also considered in the comparison. RESULTS: A more uniform coverage of the PTV in the IMRT dose plans with respect to the 3DCRT plan was found (for PTV2: V90% = 94.3 for 3DCRT, 97.6 for SS5, 98 for SS10 and 98.1 for SW; V107% = 20.7 for 3DCRT, 5.9 for SS5, 2 for SS10 and 1.3 for SW). Concerning OARs, they all present a significant reduction of mean and/or maximum dose and dose-volume patterns assessed from DVHs: in particular the mean dose of parotids decrease on average of about 13.5Gy passing from 3DCRT to IMRT with an average reduction of NTCP ranging from about 20% to more than 40% for radically treated patients, depending on the chosen end-point. IV and TV are also slightly smaller with IMRT. The results obtained with SS techniques employing 10 or more intensity levels are comparable with those obtained with SW; no differences between SS10 and SW may be appreciated when considering the DVHs of PTV, CTV and OARs. On the other hand, in some cases SS5 may be slightly sub-effective with respect to SS10 SW when considering PTV coverage and Dmax of the spinal cord. CONCLUSIONS: With the Varian planning and delivery system, Step-and-shoot approximations of inversely optimised fluences in head-neck IMRT compare well with SW delivery, even with only five intensity levels. With a number of intensity level of 10 or more, no differences can be appreciated in PTV coverage/OAR sparing with respect to SW. PMID- 16260058 TI - Inactivation of Bacillus spores by the combination of moderate heat and low hydrostatic pressure in ketchup and potage. AB - The combination effect of moderate heat and low hydrostatic pressure (MHP) on the reduction of Bacillus subtilis, Bacillus coagulans and Geobacillus stearothermophilus spores in food materials (potage and ketchup) was investigated. These bacterial spores were suspended in potage (pH 7), acidified potage (pH 4), neutralized ketchup (pH 7) and ketchup (pH 4). The suspensions were treated with and without pressure (100 MPa) and temperatures of 65-85 degrees C for 3 to 12 h. The bacterial spores were inactivated by 4-8 log cycles during MHP treatment in potage, acidified potage and ketchup, whereas the spores were highly resistant to long time heat treatment in potage and neutralized ketchup. The degrees of spore destruction were mostly dependent on pH and medium composition during MHP treatment. The inactivation effect in MHP treatment was higher at the pH 7 than at pH 4 both in ketchup and potage. The bacterial spores showed higher inactivation in potage than ketchup during MHP treatment. PMID- 16260057 TI - Prevalence, serotypes and virulence genes of Shiga toxin-producing Escherichia coli isolated from ovine and caprine milk and other dairy products in Spain. AB - The aim of this study was to determinate the prevalence, serotypes and virulence genes of Shiga toxin-producing Escherichia coli (STEC) strains isolated from different dairy products (DP) in Spain with the purpose of determining whether DP represent a potential source of STEC pathogenic for humans. A total of 502 DP were examined from 64 different ovine and caprine flocks and 6 dairy plants in Extremadura (Western Spain). Samples were collected monthly between March 2003 and June 2004 and included 360 unpasteurised milk obtained from the bulk tank, 103 fresh cheese curds and 39 cheeses. Samples obtained were examined for STEC using genotypic (PCR) methods. STEC strains were detected from 39 (10.8%) bulk tank, 4 (3.9%) fresh cheese curds and 2 (5%) cheese, whereas O157:H7 serotype were isolated from one (0.3%) bulk tank. A total of 9 STEC strains (O27:H18, O45:H38, O76:H19, O91:H28, O157:H7, ONT:H7, ONT:H9 and ONT:H21) were identified in this study. One of them, the serotype O27:H18, has not been reported previously as STEC. PCR showed that 3 strains carried stx1 genes, 5 possessed stx2 genes and 1 both stx1 and stx2. Whereas all STEC caprine isolates showed ehxA genes, only O157:H7 serotype showed eae virulence genes. The strain O157:H7 isolated possessed intimin type gamma1 and belonged to phage type 31. This study confirms that dairy product is an important reservoir of STEC pathogenic for humans. PMID- 16260059 TI - Fast-melting tablets based on highly plastic granules. AB - Highly plastic granules that can be compressed into tablets at low pressure were developed to make fast-melting tablets (FMTs) by compression method. The highly plastic granules are composed of three components: a plastic material, a material enhancing water penetration, and a wet binder. One of the unique properties of the highly plastic granules is that they maintain a porous structure even after compression into tablets. The porous and plastic nature of the granules allows fast absorption of water into the compressed tablet for fast melting/dissolution of the tablet. The prepared tablets possess tablet strength and friability that are suitable for multi-tablet packages. The three-component highly plastic granules provide an effective way of making FMTs by compression. PMID- 16260060 TI - Construction and characterization of a t-PA mutant for use in ATTEMPTS: a drug delivery system for achieving targeted thrombolysis. AB - To resolve the bleeding risk associated with thrombolytic therapy, we have designed an approach, termed ATTEMPTS (Antibody Targeted Triggered Electrically Modified Prodrug Type Strategy), to deliver t-PA to the clot site in an inactive form and then trigger its conversion to the active form, so that it would selectively activate the clot bound plasminogen while alleviating the bleeding risk. This delivery system was composed of a large protein complex, consisting of two components: (i) a heparin-modified, negatively charged fibrin-targeting antibody; and (ii) a cationic peptide-modified, positively charged t-PA. Both in vitro and in vivo studies have confirmed the feasibility of this targeted drug delivery approach. A site-specific thrombolysis was observed in animals, without concomitant depletion of the coagulation factors -- the phenomenon in conventional thrombolytic therapy that contributes to the bleeding risk. Despite promise, the chemical conjugation method employed previously in the preparation of the cationic peptide-modified t-PA also revealed several major shortcomings. The primary drawback was that the number of the cationic peptides and the location at which these peptides were attached to a t-PA molecule could not be regulated by using the chemical conjugation method. As a consequence, the resultant modified t-PA possessed a wide range of heparin-binding strength, rendering the inhibition of t-PA activity by heparin binding ineffective. In this paper, we present a new strategy in producing the desired modified t-PA, utilizing the genetic engineering approach. A computer simulation-guided rational design strategy was adopted to identify the most desirable site in t-PA (i.e. the 37-loop) for incorporation of the heparin-binding peptide sequence. By altering the amino acid composition via mutation at three locations, i.e. Ser(300) to Cys, Gly(302) to Arg, and Glu(303) to Arg, a highly cationic nanomer sequence consisting of (297)KHRRCPRRR(304) and possessing a well-demonstrated heparin binding domain was established within the 37-loop. To ensure the binding of heparin to this specifically modified domain, a cysteine residue (i.e. Cys(300)) was created to allow for site-specific conjugation of an additional heparin binding peptide (i.e. the LMWP peptide previously developed in our laboratory) to this domain via the chemical conjugation method. In vitro fibrinolysis assays showed that both the t-PA mutant and the LMWP-attached t-PA mutant exhibited a fibrinolytic potency similar to that of the wild type t-PA. Inhibition studies using small chromogenic substrates demonstrated that the activity of mutant tPA LMWP could be significantly inhibited by heparin binding. In conclusion, using computer simulation and molecular biology approaches, a mutated t-PA that meets the needs of the ATTEMPTS system, in providing a safe thrombolytic therapy, could be readily prepared. PMID- 16260062 TI - GASepo-a software solution for quantitative analysis of digital images in Epo doping control. AB - A software has been developed that is aimed at quantitative analysis of images acquired by isoelectric focusing and double blotting procedures used for recombinant erythropoietin doping control. It represents a unified and easy-to use tool for Epo doping experts in WADA accredited laboratories. It is based on image segmentation philosophy that enables identification of individual bands whose characteristics are needed for evaluation of the Epo doping positivity criteria. Several modules implemented in the GASepo software include an original know how, in particular, the method of robust calculation of the cut-off line, band segmentation and classification algorithms. GASepo is being used in several doping control laboratories worldwide. PMID- 16260061 TI - Leanness and type 2 diabetes in a population of indigenous Australians. AB - OBJECTIVE: To determine the prevalence of type 2 diabetes and its risk factors in a population of indigenous Australians. RESEARCH DESIGN AND METHODS: A cross sectional study of 332 indigenous community residents aged 15 years and over with fasting blood samples and anthropometric measurements. RESULTS: Almost half of the study population (47.3%) was extremely lean (BMI<22 kg/m(2)). Leanness was particularly pronounced in the youngest age group (15<20 years), 78% of which had a BMI<22 kg/m(2). The prevalence of diabetes was 12%. It was highest in those 45 54 years and declined in older aged people. No cases of diabetes were detected in those aged less than 30 years. Diabetes prevalence was strongly linked to BMI and age (age-adjusted odds ratio=24.1, 95% CI 6.0-96.5, p<0.001) for BMI>or=25 kg/m(2) versus BMI<22 kg/m(2). Those with the lowest diabetes risk profile are lean (BMI<22 kg/m(2)) and/or young (age 15-34 years). CONCLUSIONS: These results highlight that strategies to prevent or delay the onset of diabetes should focus on the maintenance of leanness from adolescence and throughout adult life whilst young people are still in the process of forming lifelong habits. PMID- 16260063 TI - Expressions of cytokines and chemokines in the dorsal motor nucleus of the vagus nerve after right vagotomy. AB - The aim of this study was to investigate the expression of cytokines, tumor necrosis factor alpha (TNF-alpha), interleukin-1 beta (IL-1beta), interleukin-6 (IL-6) and transforming growth factor-beta 1 (TGF-beta1) and chemokines, fractalkine, monocyte chemoattractant protein 1 (MCP-1) and stromal cell-derived factor 1 (SDF-1) in the dorsal motor nucleus of the vagus nerve (DMV) after right vagotomy. Results showed that the immunoreactivities of IL-1beta, IL-6, TGF beta1, fractalkine and MCP-1 were upregulated in the DMV at 14 days and the upregulation persisted at least until 28 days following right vagotomy. Quantification analysis revealed significant increases in the number of their immunopositive cells in the right DMV at 14 and 28 days after right vagotomy. Moreover, the upregulation of TNF-alpha immunoreactivity and significantly increased number of TNF-alpha-immunopositive cells were observed in the injured DMV at 7 and 14 days, and the increase in SDF-1-immunopositive cells at 14 days, after right vagotomy. Real time RT-PCR analysis showed the significant increase in the mRNA expression of IL-1beta, fractalkine and MCP-1 at 7 days, and the upregulation of TNF-alpha mRNA expression at 1 day after vagotomy. However, the peak increase in TGF-beta1 mRNA expression was observed at 1 day and the significant increase persisted at least until 14 days following right vagotomy. Double immunofluorescence analysis showed co-localization of lectin, a marker for microglia with CX3CR1 but not with IL-1beta at 14 days following right vagotomy. This study suggests that cytokines and chemokines involved in neuroprotection and neurodestruction could be activated in the axotomized DMV. However, it warrants further investigation to understand the neurodestructive and neuroprotective mechanisms that determine the fate of the vagal motoneurons after vagotomy. PMID- 16260064 TI - Optimization, evaluation, and characterization of molecularly imprinted polymers. AB - The underlying mechanisms for molecular recognition exhibited by the imprinting effect can be attributed to two processes. The pre-organization of complementary functional groups in the polymer by the template and the formation of a shape selective cavity that is complementary to the template. However, measurements of binding and selectivity combine all effects contributing to molecular recognition in MIPs into one figure of merit. If the two molecules being compared are not enantiomers, then there are other factors which contribute to differential binding such as size or different partitioning effects due to differences in polarity, hydrophobicity, ionization state or shape and/or conformational effects. The best probe for the imprinting effect is therefore an enantiomeric pair. Therefore, the first section of this article discusses enantioselective optimization of polymerization, the second section will review methods employed for evaluation of MIPs and the last section will cover materials science methods used to characterize the physical properties of MIP materials. PMID- 16260065 TI - Stress response and binge eating disorder. AB - In clinical practice, obese patients report stress as a primary trigger for binge eating. However, the biological mechanism underlying this relationship is poorly understood. This paper presents, a theoretical overview of how cortisol secretion, a major component of the stress response, could play a role in binge eating, given that exogenous glucocorticoids can lead to obesity by increasing food intake. I will discuss findings from recent studies demonstrating links between laboratory stress, cortisol, food intake and abdominal fat in humans. Cortisol is elevated following laboratory stressors in women with anorexia nervosa (AN), bulimia nervosa (BN), and obesity, but has not been widely studied in women with binge eating disorder (BED). Additionally, I will review recent findings demonstrating a greater cortisol response to stress in obese women with BED compared to non-BED. PMID- 16260066 TI - A key role for the microglial NADPH oxidase in APP-dependent killing of neurons. AB - Reactive oxygen species (ROS) and deposition of cleaved products of amyloid precursor protein (APP) are thought to contribute to neuronal loss observed in Alzheimer's disease (AD). The relationship between these factors was studied in a neuroblastoma and microglia co-culture system. Overexpression of wild-type APP (APP-wt) or APP with three mutations typical of familial AD (APP-3m) in SH-SY5Y neuroblastoma cells did not directly alter their morphology, growth rate, cell cycle or H(2)O(2) sensitivity. In a co-culture of APP-wt neuroblastoma cells with microglia, microglial cells generated ROS and neuronal cells died. The cell death was more pronounced in APP-3m-expressing neurons. Neuroblastoma cell death was attenuated by ROS-scavengers and was dose-dependently inhibited by the NADPH oxidase inhibitor diphenyleneiodonium chloride (DPI). Macrophage cell lines behaved similarly to microglia in the co-culture model. However, a macrophage cell line deficient in the NADPH oxidase subunit, gp91phox, failed to kill neurons. These results suggest that APP-dependent microglia activation and subsequent ROS generation by the phagocyte NADPH oxidase play a crucial role in neuronal killing in a cellular model of AD. PMID- 16260067 TI - Synthesis and antiprotozoal activity of some 2-(trifluoromethyl)-1H-benzimidazole bioisosteres. AB - A series of 2-(trifluoromethyl)-1H-benzimidazole derivatives with various 5- and 6-position bioisosteric substituents (-Cl, -F, -CF3, -CN), namely 1-7, were prepared using a short synthetic route. Each analogue was tested in vitro against the protozoa Giardia intestinalis and Trichomonas vaginalis in comparison with albendazole and metronidazole. Several analogues had IC50 values < 1 microM against both species, which make them significantly more potent than either standard. Compound 4 [2,5(6)-bis(trifluoromethyl)-1H-benzimidazole], was 14 times more active than albendazole against T. vaginalis. This compound (4) also showed moderate antimalarial activity against W2 and D6 strains of Plasmodium falciparum (5.98 and 6.12 microM, respectively). Studying further structure activity relationships through the use of bioisosteric substitution in these benzimidazolic derivatives should provide new leads against protozoal and possibly malarial diseases. PMID- 16260068 TI - Synthesis and evaluation of new quinazolone derivatives of nalidixic acid as potential antibacterial and antifungal agents. AB - In continuation of our work on synthesis of biheterocycles carrying the biodynamic heterocyclic systems at position 3, a series of new nalidixic acid derivatives having quinazolones moiety were synthesised to achieve enhanced biological activity and wide spectrum of activity. Nalidixic acid was first converted into its acid chloride using thionyl chloride as an acylating agent at laboratory temperature. Later it was converted to methyl ester. Nalidixoyl chloride formed vigorously reacts with methanol to give a methyl ester of nalidixic acid. The ester on addition of hydrazine hydrate furnished nalidixic acid hydrazide. Appropriate anthranilic acid was refluxed with acetic anhydride to form Benzoxazine/Acetanthranil. 5-iodo-derivative of anthranilic acid was prepared and also utilised to obtain 6-iodo-Benzoxazine/Acetanthranil. Also, 6 nitro-Benzoxazine/Acetanthranil was obtained by nitration of acetanthranil using conc. H(2)SO(4) and fuming HNO(3). Equimolar proportions of the appropriate synthesised acetanthranils and nalidixic acid hydrazide in the presence of ethanol were refluxed to synthesise quinazolones. Elemental analysis and IR spectra confirmed nalidixic acid hydrazide formation. The structures of the compounds obtained have been established on the basis of Spectral (IR, (1)H NMR and mass) data. The current study also involves in vitro antimicrobial screening (using Agar dilution and Punch well diffusion method) of synthesised quinazolone derivatives bearing nalidixic acid moiety on randomly collected microbial strains. The derivatives Ga (NAH), Gb (QN) and Gd (NiQNA) showed marked inhibitory activity against enteric pathogen like Aeromonas hydrophila, a causative agent of diarrhoea in both children as well as adults. Among the respiratory pathogens included in study, derivative Gd (NiQNA) was found to be active against Streptococcus pyogenes. No significant inhibitory activity was seen by any of synthesised derivatives against Coagulase negative Staphylococcus. Derivative Ga (NAH) was found to show very high activity against the Candida colonies and derivative Gd (NiQNA) was also found to exhibit inhibitory activity against Candida albicans; a normal flora of the human body which plays an important role in causing opportunistic infections in immunocompromised hosts. Proteus vulgaris, a gram-negative bacteria included in our study was found to be inhibited by derivative Gb (QN). PMID- 16260069 TI - Synthesis and biological evaluation of 1beta-methylcarbapenems having guanidino moieties. AB - The synthesis of a new series of 1beta-methylcarbapenems having the substituted guanidinocarbonyl pyrrolidine moieties is described. Their in vitro antibacterial activities against both Gram-positive including MRSA and Gram-negative bacteria were tested and the effect of substituents on the pyrrolidine ring was investigated. In particular, the compound Ib having piperazinylguanidine moiety showed the most potent antibacterial activity. PMID- 16260070 TI - [Influenza vaccination among health-care workers. Vaccination rates in universitary hospital of Besancon, winter 2003-2004]. AB - BACKGROUND: Influenza vaccination reduces the mortality of the patients when the vaccination rates of healthcare workers is important. PURPOSE AND METHOD OF THE STUDY: To investigate the vaccination rates at the Universitary Hospital of Besancon by anonymous questionnaire. RESULTS: Three thousand hundred seventy seven answers were analyzed (228 men and 1145 women). Two hundred seventy-seven persons declared receiving the vaccine (20.1%) corresponding to sixty-three men (27.6%) and two hundred thirteen women (18.6%) (P = 0.001). The average age of the healthcare workers vaccined was of 38.9+/-11 years. Among most than 50 years, 34% was vaccined. Among the doctors, 40.5% were vaccined against 20.6% of the nurses. In the services of geriatrics, 78.5% of the staff was vaccined. CONCLUSIONS: Our results indicate a weak rate of influenza vaccination in our establishment and a misunderstanding of the character nosocomial of the influenza among the nurse. PMID- 16260071 TI - [Lymphocytic colitis following administration of mianserine]. PMID- 16260072 TI - Age-related changes in the immune response to influenza vaccination in a racially diverse, healthy elderly population. AB - Proliferation to phytohemagglutinin (PHA) and cell-mediated responses to influenza vaccine (FLU) were assessed in healthy elderly Caucasians, African Americans, Latinos and young Caucasians before and after immunization. Both PHA- and FLU-induced proliferation were reduced in elderly subgroups. FLU-induced proliferation increased post-vaccination in all elderly, except African Americans, but responses were lower than young. IL-2 and IFN-gamma levels did not change after vaccination of elderly, regardless of race. The altered response to FLU in elderly African Americans, compared to elderly Caucasians and Latinos, indicates that racial background is important when drawing conclusions about immune responses to FLU in mixed elderly populations. PMID- 16260073 TI - Simultaneous detection of antibodies to foot-and-mouth disease non-structural proteins 3ABC, 3D, 3A and 3B by a multiplexed Luminex assay to differentiate infected from vaccinated cattle. AB - For the first time, a multiplex bead immunoassay was used to test simultaneously, with a single sample, the immune response to foot-and-mouth disease non structural proteins 3ABC, 3A, 3B and 3D from experimentally infected and vaccinated cattle. We cloned and expressed these non-structural proteins (NSPs) as recombinant antigens. The purified proteins were coupled to microspheres labeled with anti-His monoclonal antibody with different proportions of red and orange fluorescent dyes and reacted against serum specimens. Antibody reacting against different NSPs, and thus, the different colored beads, was detected by use of the Luminex system. This multiplex bead immunoassay can detect the immune response to NSPs in cattle as early as 7 days post-infection. In general antibodies to the protein 3D appeared early after infection and anti-3ABC antibodies were detected at higher levels than the other NSPs. A clear differentiation was established between infected and vaccinated or uninfected cattle. The multiplex bead immunoassay was compared to individual indirect enzyme linked immunosorbent assays (iELISAs) for the same NSP's responses. Results indicated that this new assay had a high positive correlation with those generated by iELISA. The Luminex-based technology promises to be a sensitive and efficient method that permits multiplexed NSP antibody detection from a single sample and would therefore provide both a time and cost saving to the laboratory. PMID- 16260074 TI - Functional aspects of cross-legged sitting with special attention to piriformis muscles and sacroiliac joints. AB - BACKGROUND: Transversely oriented pelvic muscles such as the internal abdominal oblique, transversus abdominis, piriformis and pelvic floor muscles may contribute to sacroiliac joint stability by pressing the sacrum between the hipbones. Surface electromyographic measurements showed that leg crossing lowers the activity of the internal oblique abdominal muscle significantly. This suggests that leg crossing is a substitute for abdominal muscle activity. No previous studies addressed piriformis muscle and related pelvic structures in cross-legged sitting. METHODS: Angles of pelvis and femur were measured in healthy subjects in standing, normal sitting and cross-legged sitting, and were used to simulate these postures on embalmed pelvises and measure piriformis muscle elongation. Deformations of pelvic ring and iliolumbar ligament caused by piriformis muscle force were measured on embalmed pelvises. FINDINGS: Cross legged sitting resulted in a relative elongation of the piriformis muscle of 11.7% compared to normal sitting and even 21.4% compared to standing. Application of piriformis muscle force resulted in inward deformation of the pelvic ring and compression of the sacroiliac joints and the dorsal side of the pubic symphysis. INTERPRETATION: Cross-legged sitting is common. We believe that it contributes to sacroiliac joint stability. This study demonstrates the influence of the piriformis muscle on sacroiliac joint compression. The elongation of the piriformis muscle bilaterally by crossing the legs may be functional in the build up of active or passive tension between sacrum and femur. PMID- 16260075 TI - Wear patterns on tibial plateau from varus osteoarthritic knees. AB - BACKGROUND: The knowledge of cartilage wear patterns at the medial tibial plateau is important to understand the main causes of arthritis in varus knees. The most important factors influencing knee arthritis in fact seem to be the severity of the degenerative changes determined by the lower limb mechanical axis and the abnormal knee joint kinematics which frequently results from dysfunction of the anterior cruciate ligament. METHODS: We studied the wear patterns of cartilage damage in 70 medial tibial plateaus resected at operation during total knee arthroplasty indicated for varus osteoarthritic knee. Anterior cruciate ligament and medial meniscus integrity was assessed intra-operatively. Calibrated digital images were used to measure the wear patterns with a standard software tool. The medial compartment of the tibial plateau was divided into six zones, and the amount of cartilage and bone destruction in each zone was classified into two grades. FINDINGS: The wear pattern was found to be highly dependent upon knee varus deformity (Mann Whitney P<0.001) and anterior cruciate ligament integrity (Friedman P<0.0005). Anterior cruciate ligament was found intact in 35.7% of the cases. Wear patterns on intact anterior cruciate ligament knees occurred in the central to medial aspect of the tibial plateau. Anterior cruciate ligament deficient knees had significantly larger wear patterns anteriorly and posteriorly in the most medial region of the medial plateau. INTERPRETATION: These observations suggest altered joint mechanics exist in anterior cruciate ligament deficient varus knees, which would worsen cartilage degeneration and osteoarthritis progression. PMID- 16260076 TI - Organochlorine contaminant and retinoid levels in blubber of common dolphins (Delphinus delphis) off northwestern Spain. AB - The effect of age, sex, nutritive condition and organochlorine concentration on blubber retinoid concentrations was examined in 74 common dolphins incidentally caught off northwestern Spain. Age and blubber lipid content were strong determinants of the retinoid concentrations in males, while these variables did not account for the variation found in females. Retinoids were positively correlated with organochlorines in males and negatively in females. However, pollution levels were moderate and likely to be below threshold levels above that a toxicological response is to be expected. Thus, a cause-effect relationship between organochlorine and retinoid concentrations could not be properly established, and the observed correlation may be the result of an independent association of the two variables with age. Further research on the influence of the best predictor variables on retinoid dynamics is required to implement the use of retinoids as biomarkers of pollutant exposure in cetaceans. PMID- 16260077 TI - Neuronal control of leech behavior. AB - The medicinal leech has served as an important experimental preparation for neuroscience research since the late 19th century. Initial anatomical and developmental studies dating back more than 100 years ago were followed by behavioral and electrophysiological investigations in the first half of the 20th century. More recently, intense studies of the neuronal mechanisms underlying leech movements have resulted in detailed descriptions of six behaviors described in this review; namely, heartbeat, local bending, shortening, swimming, crawling, and feeding. Neuroethological studies in leeches are particularly tractable because the CNS is distributed and metameric, with only 400 identifiable, mostly paired neurons in segmental ganglia. An interesting, yet limited, set of discrete movements allows students of leech behavior not only to describe the underlying neuronal circuits, but also interactions among circuits and behaviors. This review provides descriptions of six behaviors including their origins within neuronal circuits, their modification by feedback loops and neuromodulators, and interactions between circuits underlying with these behaviors. PMID- 16260079 TI - Relationship between preferential interaction of a protein in an aqueous mixed solvent and its solubility. AB - The present paper is devoted to the derivation of a relation between the preferential solvation of a protein in a binary aqueous solution and its solubility. The preferential binding parameter, which is a measure of the preferential solvation (or preferential hydration) is expressed in terms of the derivative of the protein activity coefficient with respect to the water mole fraction, the partial molar volume of protein at infinite dilution and some characteristics of the protein-free mixed solvent. This expression is used as the starting point in the derivation of a relationship between the preferential binding parameter and the solubility of a protein in a binary aqueous solution. The obtained expression is used in two different ways: (1) to produce a simple criterion for the salting-in or salting-out by various cosolvents on the protein solubility in water, (2) to derive equations which predict the solubility of a protein in a binary aqueous solution in terms of the preferential binding parameter. The solubilities of lysozyme in aqueous sodium chloride solutions (pH=4.5 and 7.0), in aqueous sodium acetate (pH=8.3) and in aqueous magnesium chloride (pH=4.1) solutions are predicted in terms of the preferential binding parameter without any adjustable parameter. The results are compared with experiment, and for aqueous sodium chloride mixtures the agreement is excellent, for aqueous sodium acetate and magnesium chloride mixtures the agreement is only satisfactory. PMID- 16260078 TI - Protective medical legislation deficient knowledge of maternity (health and safety) rights for work-adjustment exists amongst flexible trainee doctors: is there a risk to maternal and foetal health? AB - OBJECTIVE: To assess awareness, identify knowledge source and evaluate uptake amongst doctors of "health and safety rights" (HSR) contained within the current European protective medical legislation for pregnant workers. STUDY DESIGN: A descriptive cross-sectional pilot study, by a postal questionnaire during the period 1998-1999, targeted 97 UK doctors (West Midlands region) after their first pregnancy. RESULTS: Of 67 respondents (response rate 73%), 41 (61%) were Registrars (SpR) and 25 (37%) SHOs: 80% work-schedules did not change during pregnancy. Only 11% (95% CI, 4-21%) of the doctors surveyed actually knew their maternity rights. 66.2% had no knowledge of maternity legislation; 80% of respondents had not taken up health and safety rights. Fifty-two percent (95% CI, 40-65%) reported maternal and neonatal complications. CONCLUSIONS: In a self selected group of flexible trainees following their first pregnancy, only one in five female doctors have adequate knowledge about the legislative "health and safety rights" of work-schedule adjustment. A combination of reasons may contribute to the low uptake of these rights. The question of whether or not poor knowledge and uptake of legislative rights may be detrimental towards pregnancy and neonatal complications requires a large prospective study. An improvement in the knowledge of current maternity legislation could occur by targeting all medical students, all doctors, postgraduate trainers and National Health Service (NHS) employers. PMID- 16260081 TI - The First 30 Years of EUROPEAN UROLOGY (1975-2005). PMID- 16260082 TI - Re: Roddam AW, Duffy MJ, Hamdy FC, et al. Use of prostate-specific antigen (PSA) isoforms for the detection of prostate cancer in men with a PSA Level of 2-10 ng/ml: systematic review and meta-analysis. Eur Urol 2005;48:386-99. PMID- 16260083 TI - KAI1/CD82, a tumor metastasis suppressor. AB - Tetraspanin KAI1/CD82 is a wide-spectrum tumor metastasis suppressor. KAI1/CD82 suppresses tumor metastasis by primarily inhibiting cancer cell motility and invasiveness. In tetraspanin-enriched microdomain, KAI1/CD82 associates with the proteins important for cell migration such as cell adhesion molecule, growth factor receptor, and signaling molecule. Likely, KAI1/CD82 down-regulates the functions of these motility-related proteins to inhibit cell migration. The loss of KAI1/CD82 expression in invasive and metastatic cancers is due to a complex, epigenetic mechanism that probably involves transcription factors such as NFkappaB, p53, and beta-catenin. PMID- 16260084 TI - Improved and selective platinum recovery from spent alpha-alumina supported catalysts using pretreated anionic ion exchange resin. AB - Improved and selective recovery of platinum from a spent dehydrogenation platinum alpha-alumina supported catalyst using a strong basic ion exchange resin is reported. Platinum and other precious metal group (PMG) complexes are leached using concentrated hydrochloric acid along with about 0.20 vol.% nitric acid as an oxidizing agent from de-coked and crushed spent catalyst. Effects of hydrochloric acid concentration, time, and temperature in leaching stage are investigated. The strong basic anionic resin is treated by sodium hydroxide solution to replace chloride anion by hydroxyl group ion. The supernatant of the leaching process is passed through a fixed column of hydroxylated strong base anionic resin. The treated resin on which the platinum complex is adsorbed is dried and burned in an oxidizing atmosphere at 750-800 degrees C. The recovered gray metallic powder is mainly platinum. Results compared with those obtained from untreated anionic resin show that adsorption of platinum complexes onto the treated anionic resin is more selective and the yield of separation is considerably improved. The breakthrough curves of the pretreated anion exchanger and that of untreated exchange resin reveals that the capacity of the hyroxilated resin is decreased by about 14%. These breakthrough curves can be used for calculation of height of a practical exchange plate (HPEP) for design purposes. PMID- 16260085 TI - Solid/solution partitioning and speciation of heavy metals in the contaminated agricultural soils around a copper mine in eastern Nanjing city, China. AB - Solid/solution partition coefficient (Kd) and speciation of soil heavy metals can be used for predicting their environmental risks. The Kd values and solution speciation of soil Cu, Cd and Zn were analyzed in 40 samples of contaminated agricultural soils around Jiuhua copper mine in eastern Nanjing city, China. The Kd ranges (and mean values) for soil Cu, Cd and Zn are 703-7418 (3453), 37.3-3963 (940) and 319-17965 (7244) L kg-1, respectively, showing a large variability both for metals and soils. The results of differential pulse anodic stripping voltammetry (DPASV) indicates that 95.6% solution Cu is bound to dissolved organic ligands. About half of the dissolved Zn is DPASV-labile at pH<6, while 92.1% solution Zn is in the form of organic complexes at pH>6. DPASV-labile Cd is ranged from 22.6 to 98.7% with the mean value of 56.3%. Multiple linear regressions indicate that Kd, the dissolved and DPASV-labile concentrations of Cd and Zn are mostly influenced by the soil solution pH with R2 of 0.50, 0.59 and 0.63, respectively for Cd, and 0.58, 0.72 and 0.64, respectively for Zn. Considering the second parameter of corresponding soil metal, the linear relationships of Kd with pH were improved with R2 of 0.70 and 0.73 for Cd and Zn, respectively. However, the solubility of soil Cu was insensitive to pH. Only SOC shows a weak relationship to the dissolved Cu with R2 of 0.21. As for its Kd, total soil Cu is the most significant factor. But for DPASV-labile Cu, no soil parameters were found to be good predictors. PMID- 16260086 TI - Nicotine enhances both foreground and background contextual fear conditioning. AB - The present study examined if nicotine enhances contextual fear conditioning when the training context is either a background stimulus or a foreground stimulus. In the background conditioning experiment, mice were trained using two auditory conditioned stimulus (CS; 30 s, 85 dB white noise)-footshock unconditioned stimulus (US; 2 s, 0.57 mA) pairings and tested 24 h later. In the foreground conditioning experiment, mice were trained with two presentations of a footshock US (2 s, 0.57 mA) and tested 24 h later. Mice received 0.09 mg/kg nicotine before training and testing. For both the foreground and background conditioning experiments, nicotine enhanced contextual conditioning. No enhancement of the auditory CS-US association was seen. These results demonstrate that nicotine enhances contextual fear conditioning regardless of whether the context is a background stimulus or a foreground stimulus during conditioning. PMID- 16260087 TI - Changes in neuromuscular function after tasks involving control of EMG versus torque feedback of the same duration. AB - This study was designed to compare alterations in neuromuscular function after two tasks of similar duration involving the control of (1) torque level fixed at 40% maximal voluntary contraction (MVC) torque (torque task) and (2) EMG level when exerting 40% MVC torque on the knee extensor muscles. Ten healthy subjects volunteered to participate in two testing sessions separated by approximately 2 h. Contraction duration for the EMG task was fixed for each subject to the time to task failure of the torque task (104+/-20s). MVC, maximal voluntary activation level, muscle compound action potential (M-wave), peak twitch and potentiated peak doublet were assessed before and immediately after each task using electrical stimulation of the femoral nerve. Average EMG activity of quadriceps muscle increased (p<0.01) during the torque task from 27.7+/-5.4% to 46.2+/-19.3% maximal EMG, whereas torque decreased during the EMG task from 41.5+/-2.9% to 28.9+/-3.8% MVC torque. Alterations in MVC torque (p<0.01) and maximal voluntary activation level (p<0.05) were comparable at termination of the two tasks. Rate of perceived exertion was greater (p<0.05) at the end of the torque task compared to the EMG task. Despite the absence of change in the M-wave for either task, potentiated peak doublet was altered after the torque task (-18+/-14%, p<0.01), whereas there was no change after the EMG task (p>0.05). The absence of peripheral failure at the end of the EMG task could be attributed to (1) a lower intramuscular pressure allowing a lesser accumulation of metabolites and (2) a slower rate of PCr hydrolysis compared to the torque task. PMID- 16260088 TI - Okadaic acid induces JNK activation, bim overexpression and mitochondrial dysfunction in cultured rat cortical neurons. AB - Apoptosis via tau phosphorylation has been implicated in the selective neuronal losses seen in Alzheimer's disease (AD). Previous studies in vivo and in cultured neurons have shown that okadaic acid (OA) evokes tau phosphorylation to initiate a neurodegeneration that resembles the pathogenesis of AD. In an effort to identify additional key molecules in this neurodegeneration, we treated cultured rat neurons with OA and examined the apoptosis-related effects, such as changes in mitochondrial activity and expression levels of JNK, Bim, Bad, Bax and caspase 3. Western blotting revealed that phosphorylation of JNK and c-jun occurred first, followed by increased expression of Bim and subsequent caspase-3 activation in OA-treated neurons. In contrast, Bad levels decreased as early as 4 h after OA treatment. Immunocytochemistry showed that the increased phospho-JNK immunoreactivity was localized in the cytosol of degenerating neurons, while increased phospho-c-jun was localized in the nucleus. The mitochondria showed decreased membrane potential and increased swelling after OA treatment. Collectively, these data suggest that JNK- and Bim-related mitochondrial dysfunction is involved in OA-induced neurodegeneration. PMID- 16260089 TI - Benfotiamine alleviates diabetes-induced cerebral oxidative damage independent of advanced glycation end-product, tissue factor and TNF-alpha. AB - Diabetes mellitus leads to thiamine deficiency and multiple organ damage including diabetic neuropathy. This study was designed to examine the effect of benfotiamine, a lipophilic derivative of thiamine, on streptozotocin (STZ) induced cerebral oxidative stress. Adult male FVB mice were made diabetic with a single injection of STZ (200 mg/kg, i.p.). Fourteen days later, control and diabetic (fasting blood glucose >13.9 mM) mice received benfotiamine (100 mg/kg/day, i.p.) for 14 days. Oxidative stress and protein damage were evaluated by glutathione/glutathione disulfide (GSH/GSSG) assay and protein carbonyl formation, respectively. Pro-oxidative or pro-inflammatory factors including advanced glycation end-product (AGE), tissue factor and tumor necrosis factor alpha (TNF-alpha) were evaluated by immunoblot analysis. Four weeks STZ treatment led to hyperglycemia, enhanced cerebral oxidative stress (reduced GSH/GSSG ratio), elevated TNF-alpha and AGE levels without changes in protein carbonyl or tissue factor. Benfotiamine alleviated diabetes-induced cerebral oxidative stress without affecting levels of AGE, protein carbonyl, tissue factor and TNF-alpha. Collectively, our results indicated benfotiamine may antagonize diabetes-induced cerebral oxidative stress through a mechanism unrelated to AGE, tissue factor and TNF-alpha. PMID- 16260090 TI - Garcinone B reduces prostaglandin E2 release and NF-kappaB-mediated transcription in C6 rat glioma cells. AB - In the course of our survey of natural compounds inhibiting prostaglandin E2 release and/or lipopolysaccharide (LPS)-induced transcriptional stimulation via NF-kappaB, a central regulator of inflammatory genes, from natural resources, we found garcinone B, a xanthone from callus tissue culture of Hypericum patulum, as a compound with such pharmacological activities, that is a derivative of gamma mangostin which potently inhibits COX-1 and COX-2 activities to reduce PGE2 release from C6 rat glioma cells, and inhibits IKK activity to prevent NF-kappaB dependent COX-2 gene transcription. Garcinone B, to a lesser extent, reduced A23187-induced increase in prostaglandin E2 release than gamma-mangostin and its structurally related compound, patulone, in C6 cells. This compound also prevented LPS-induced stimulation of NF-kappaB-dependent transcription. These results suggest that garcinone B becomes a unique pharmacological tool to investigate intracellular signaling pathways involved in inflammation. PMID- 16260091 TI - Cell size-dependent Nogo-A expression in layer V pyramidal neurons of the rat primary somatosensory cortex. AB - Nogo-A mRNA and protein are present in the perikarya of neurons in both the intact and injured CNS. The present study focused primarily on Nogo-A protein expression in the primary somatosensory cortex of the adult rat. Coronal brain sections were probed with double-immunofluorescent labeling against Nogo-A together with NeuN, RBPC, or MAP-2 for confocal imaging. The sizes of the cell somata in pyramidal neurons and the thicknesses of neurites were measured on the captured confocal images. Nogo-A was expressed in larger pyramidal neurons and thicker neurites in layer V, but not in smaller pyramidal neurons and thinner neurites. Considering the morphological properties and the cell soma size reported in previous studies together with the present data, Nogo-A-positive neurons of layer V appear to be intrinsically bursting neurons that project axons to the subcortical regions. This suggests that intraneuronal Nogo-A may play roles in neurite growth and axonal regeneration of the corticofugal neurons, but not of columnar intrinsic neurons, in layer V of the S1 barrel cortex. Additionally, this study demonstrates a novel result, which is that layer V pyramidal neurons of the S1 barrel cortex exhibit a pattern of cell size dependent intraneuronal Nogo-A expression. PMID- 16260093 TI - New aliphatic nitro-compounds from Indigofera carlesii. AB - An investigation of aliphatic nitro-compounds in Indigofera carlesii resulted in the identification of two blended novel compounds: 2-O-acryl-3,6-di-O-(3 nitropropanoyl)-alpha-d-glucopyranose and 6-O-acryl-2,3-di-O-(3-nitropropanoyl) alpha-d-glucopyranose. The structures of the new compounds were elucidated on basis of spectral analysis. PMID- 16260092 TI - Neutrophil activation induced by the lectin KM+ involves binding to CXCR2. AB - The lectin KM+ from Artocarpus integrifolia, also known as artocarpin, induces neutrophil migration by haptotaxis. The interactions of KM+ with both neutrophils and the extracellular matrix depend on the lectin's ability to recognize mannose containing glycans. In the present study, we characterized the binding of KM+ to human neutrophils and the responses stimulated by this binding. Exposure to KM+ results in cell polarization, formation of a lamellipodium, and induction of deep ruffles on the cell surface. By fluorescence microscopy, we observed that KM+ is distributed homogeneously over the cell surface. KM+/ligand complexes are rapidly internalized, reaching maximum intracellular concentrations at 120 min, and decreasing thereafter. Furthermore, KM+ binding to the surface of human neutrophils is inhibited by the specific sugars, d-mannose or mannotriose. KM+ induced neutrophil migration is inhibited by pertussis toxin as well as by inhibition of CXCR2 activity. These results suggest that the KM+ ligand on the neutrophil surface is a G protein-coupled receptor (GPCR). The results also suggest that neutrophil migration induced by KM+ involves binding to CXCR2. PMID- 16260094 TI - Sleep, sleep-dependent procedural learning and vigilance in chronic cocaine users: Evidence for occult insomnia. AB - Sleep disturbance has been implicated in cocaine use; however, the nature of the disturbance and its potential effects on cognition and learning are largely unknown. Twelve chronic cocaine users completed a 23-day inpatient study that included randomized, placebo-controlled, cocaine self-administration sessions. Six subjects received cocaine on each of days 4-6 and placebo on days 18-20, the other six received cocaine on each of days 18-20 and placebo on days 4-6. Sleep was measured by polysomnography, the Nightcap sleep monitor, and self-reported measures. Simple and vigilance reaction times were measured daily; a motor sequence test of procedural learning was administered four times. Electrophysiological measures of sleep showed a different pattern than self reported sleep across cocaine administration and abstinence: total sleep time and sleep latency were at their worst by 14-17 days of abstinence while self-reported sleep was at its best. Vigilance correlated positively with electrophysiologically measured sleep and negatively with self-reported measures. Similarly, sleep-dependent procedural learning correlated with total sleep time and was impaired at 17 days abstinence relative to 2- and 3-days abstinence. Slow wave activity was lowest at days 4-9 of abstinence and highest during use and days 10-17 of abstinence. With sustained abstinence, chronic cocaine users exhibit decreased sleep, impaired vigilance and sleep-dependent procedural learning, and spectral activity suggestive of chronic insomnia. However, they report subjectively improving sleep, indicating they are unaware of this "occult" insomnia. These results suggest the possibility of homeostatic sleep drive dysregulation in chronic cocaine users. PMID- 16260095 TI - Polyhydroxyalkanoate accumulating diversity of Pseudomonas species utilising aromatic hydrocarbons. AB - A number of Pseudomonas strains accumulated polyhdroxyalkanoate (PHA) from a variety of aromatic hydrocarbons. In many strains the level of PHA accumulation was dependent on the side chain length of the phenylalkanoic acid provided for growth. 4 of the 8 strains accumulated increased levels of PHA as the side chain length of the phenylalkanoic acid substrate increased. PHA accumulated from styrene and phenylacetic acid was composed of aliphatic monomers only. The PHA accumulated from any one of the phenylalkanoic acids with 5 carbons or more in their side chain (n>or=5) was almost identical for all strains with PHA composed of both aromatic and aliphatic monomers. The predominant monomers accumulated were 3-hydroxyphenylvaleric acid and 3-hydroxyphenylhexanoic acid. The addition of the metabolic pathway inhibitors acrylic acid and 2-bromoctanoic acid resulted in decreased levels of PHA from phenylacetic acid, suggesting a role for both beta-oxidation and fatty acid synthesis in PHA accumulation from phenylacetic acid. PMID- 16260096 TI - Synthesis of a small, cysteine-rich, 29 amino acids long peptide in Mycoplasma pneumoniae. AB - A 205-210 bases long, small RNA (MP200RNA) of Mycoplasma pneumoniae encodes an open reading frame (ORF pmp200) that has the potential to be translated into a 29 amino acids long peptide with nine cysteines. The expression of this peptide in M. pneumoniae was proven indirectly by constructing a gene fusion between the ORF pmp200 and mrfp1, the gene encoding the monomeric red fluorescent protein. The fusion construct was translated in M. pneumoniae. The corresponding fusion protein, with a molecular mass of approximately 35,000 Da, was isolated and the correct sequence was proven by Edman degradation and by mass spectrometry. PMID- 16260097 TI - Human and murine paraoxonase 1 are host modulators of Pseudomonas aeruginosa quorum-sensing. AB - The pathogenic bacterium Pseudomonas aeruginosa uses acyl-HSL quorum-sensing signals to regulate genes controlling virulence and biofilm formation. We found that paraoxonase 1 (PON1), a mammalian lactonase with an unknown natural substrate, hydrolyzed the P. aeruginosa acyl-HSL 3OC12-HSL. In in vitro assays, mouse serum-PON1 was required and sufficient to degrade 3OC12-HSL. Furthermore, PON2 and PON3 also degraded 3OC12-HSL effectively. Serum-PON1 prevented P. aeruginosa quorum-sensing and biofilm formation in vitro by inactivating the quorum-sensing signal. Although 3OC12-HSL production by P. aeruginosa was important for virulence in a mouse sepsis model, Pon1-knock-out mice were paradoxically protected. These mice showed increased levels of PON2 and PON3 mRNA in epithelial tissues suggesting a possible compensatory mechanism. Thus, paraoxonase interruption of bacterial communication represents a novel mechanism to modulate quorum-sensing by bacteria. The consequences for host immunity are yet to be determined. PMID- 16260098 TI - Tuber borchii mycelial protoplasts isolation, characterization and functional delivery of liposome content, a new step towards truffles biotechnology. AB - The filametous ascomycete Tuber borchii is a plant-symbiotic ectomycorrhizal microrganism with an high value due to the production of hypogeous fruitbodies (truffles). The present work was undertaken to develop a procedure for the release of T. borchii viable protoplasts from Tuber mycelium, isolate ATTC 96540; several factors which affect the isolation, morphology and viability were examined and developed in order to improve applications of T. borchii protoplasts in morphological, biochemical and genetic investigations (protoplast fusion or transformation). Functional delivery of liposome content into T. borchii protoplasts has also been examined with a cytotoxic ribosome inactivator as saporin. T. borchii protoplasts incubation/fusion with saporin containing liposomes were made to demonstrate the absence of cell wall of 16 days cultured protoplasts. PMID- 16260099 TI - Application of liquid chromatography-mass spectrometry to biomonitoring of exposure to industrial chemicals. AB - Recent advances on biomarker research are discussed, primarily relying on experience gained with technologies based on liquid chromatography-tandem mass spectrometry (LC-MS-MS). Determination of urinary metabolites of industrial chemicals (n-hexane, benzene, toluene, and styrene) in samples from occupationally exposed workers and controls was performed by LC-MS-MS with either electrospray (ESI) or atmospheric pressure chemical ionization (APCI), as appropriate. Both phase I and II metabolites (glucuronides, sulfates, and mercapturic acids) can be detected with little or no sample manipulation, thus allowing the identification of a number of artifacts and "new" metabolites. However, experimental evidence indicates the need for properly addressing the matrix effect, which is always associated with the analysis of biological samples. Both efficient sample preparation and the use of isotopically labeled internal standards seem to be necessary to develop validated quantitative methods. PMID- 16260100 TI - Isolation and characterization of heparin and gelatin binding buffalo seminal plasma proteins and their effect on cauda epididymal spermatozoa. AB - Seventy semen ejaculates were obtained from 14 Murrah buffalo bulls and were subjected to plasma separation immediately after collection by centrifugation at 2000 rpm for 20 min and stored in liquid nitrogen until analysis. In the seminal plasma the total protein concentration were estimated and the heparin and gelatin binding (HB and GB) proteins were isolated using heparin and gelatin affinity column chromatography. The molecular weight of individual isolated HB and GB protein was determined by SDS-PAGE analysis. Buffalo bull spermatozoa was collected from cauda epididymis under aseptic conditions and was used for the in vitro fertility tests (i.e. bovine cervical mucus penetration test (BCMPT) and hypo-osmotic swelling test (HOST)). The heparin and gelatin binding buffalo seminal plasma proteins were used in six concentrations i.e. 10, 20, 30, 40, 50 and 60 microg/ml to test their effect on in vitro fertility assessment of cauda epididymal spermatozoa. The overall mean values of total protein, HB and GB proteins were recorded as 29+/-2.7, 2.61 and 0.2mg/ml, respectively. Eighteen total protein bands were observed in the range of 12-127 kDa. Eight major HB proteins were isolated in the range of 13-71 kDa. Seven major GB proteins were isolated in the range of 13-61 kDa in the buffalo seminal plasma. The mean penetration distance (mm) travelled by the buffalo cauda spermatozoa was maximum in HB proteins (26.9+/-0.6) followed by GB proteins (25.4+/-0.6) and control (21.2+/-1.4). The difference in BCMPT values between protein treated and control group was significant (P<0.05). Almost similar trend in the effect of protein on values of HOST percentage in both HB and GB proteins treated semen samples were recorded (66.4+/-0.65 and 66.1+/-0.6, respectively). The difference in HOST values between proteins treated and control group (50.4+/-2.0) was significant (P<0.05). The present results indicate that among the isolated proteins, 4 proteins were commonly seen in both the heparin and gelatin-sepharose affinity column chromatography, and the addition of buffalo seminal plasma proteins improved the in vitro sperm functions (40 microg/ml gave best results) of buffalo cauda spermatozoa. PMID- 16260101 TI - Alcohol, caffeine and smoking in relation to age at menopause. AB - OBJECTIVES: Exposures which might influence age at natural menopause have been extensively studied but, with the exception of cigarette smoking, results have been inconsistent. We sought to determine: (i) whether alcohol and caffeine intake are associated with age at menopause; (ii) whether the association of cigarette smoking with age at menopause is confined to current smokers. METHODS: Analyses drew on longitudinal data from 494 women, aged 44-60 in 1993, of whom 159 experienced menopause before intake or during follow-up. We used parametric logistic survival analysis to estimate shifts in median age at menopause for women who drink alcohol or caffeine or who smoke cigarettes. RESULTS: The estimated median age at menopause was 2.2 (95% confidence interval (CI) 0.5, 3.9) years later for women who drank alcohol 5-7 days/week (13% of the sample) than for women who did not drink alcohol (54%). For women who drank at least 1 day/week, the estimated shift was 1.3 (95% CI 0.2, 2.3) years. Caffeine intake in approximate quartiles of mg/day-0-100, >100-200, >200-400, 400+-was not related to age at menopause. Current smokers of 14+cigarettes/day (6%) experienced menopause 2.8 (95% CI -4.8, -0.8) years earlier than women who never smoked (51%). Current smokers of 1-13 cigarettes/day (5%) and former smokers (38%) experienced menopause at about the same age as women who never smoked. CONCLUSIONS: The results are compatible with a pro-estrogenic effect of moderate alcohol intake and an anti-estrogenic effect of current cigarette smoking of 14+cigarettes/day. PMID- 16260102 TI - Transfollicular drug delivery--is it a reality? AB - Once regarded as merely evolutionary remnants, the hair follicles and sebaceous glands are increasingly recognised as potentially significant elements in the percutaneous drug delivery paradigm. Interest in pilosebaceous units has been directed towards their use as depots for localised therapy, particularly for the treatment of follicle-related disorders such as acne or the alopecias. Furthermore, considerable attention has also been focused on exploiting the follicles as transport shunts for systemic drug delivery. This paper reviews various key facets of this field including; relevant aspects of pilosebaceous anatomy and physiology, the design and efficacy of follicle-targeting formulations and the emergence of quantitative modeling systems. Several novel developments in this area promise to greatly expand our understanding of this field in the near future. PMID- 16260103 TI - Effects of citrus fruit juices on cytotoxicity and drug transport pathways of Caco-2 cell monolayers. AB - The aim of this study was to correlate the taxonomy of grapefruit, pummelo, orange, lime and lemon with fruit juice-mediated cytotoxicity, modulation of epithelial permeability and P-glycoprotein (P-gp)-mediated efflux using 0-50% juice concentrations. Lime and lemon juices at 30% enhanced the absorption of [14C]-mannitol across Caco-2 cell monolayers by six- and eight-fold, respectively, but grapefruit and pummelo juices did not modulate the paracellular [14C]-mannitol transport even at 50%. Orange juice at 30% increased mannitol absorption to a comparable level as lime juice, but had minimal effects on TEER. All five juices did not modulate the passive diffusional pathway as exemplified by their negligible effects on [3H]-propranolol absorption. Grapefruit, pummelo and orange juices showed P-gp inhibitory activity by reducing rhodamine-123 (R 123) efflux and elevating R-123 cellular accumulation, but lime and lemon juices did not. Lime and lemon juices at >or=30% were cytotoxic towards Caco-2 cells. Grapefruit and pummelo juices at 10% did not affect Caco-2 cell viability, but they enhanced cell growth at concentrations of >or=30%. Orange juice increased cell viability only at lower concentrations. On the basis of these data, lime and lemon juices could be regarded as a group distinct from grapefruit and pummelo juices, while orange juice appeared to belong to a bridging group. This grouping was consistent with the categorization of the citrus fruits according to their dominant flavonoid pattern and taxonomy. PMID- 16260105 TI - The influence of the type of accelerant, type of burned material, time of burning and availability of air on the possibility of detection of accelerants traces. AB - The presented research consisted in a series of field experiments and laboratory analyses of obtained samples. Their aim was to investigate how different factors influence the possibility of identification of accelerant traces in conditions that are as similar as possible to those of real fires. The studied factors were: type of burned material, type of accelerant, length of time between lighting and extinguishing of fire and the air availability level. The obtained results show that, among investigated factors, the type of burned material has the greatest influence. The other factors, with regard to their influence, could be ranked in descending order as follows: 'other' (hard to determine and regulate factors), type of accelerant, time of burning and air availability. 'Other' factors include arrangement of the burned material and dispersion of the accelerant. PMID- 16260104 TI - Release of bioactive BMP from dextran-derived microspheres: a novel delivery concept. AB - Recent developments of biotechnology have produced a great variety of protein and bioactive drugs. For these drugs to be used therapeutically, suitable drug delivery systems have become increasingly essential. Dextran-derived biomaterials have been considered to be compatible matrices for protein and bioactive drugs because of their hydrophilic properties and ability to control drug dissolution and permeability. A novel class of dextran-glycidylmethacrylate (Dex GMA)/poly(ethylene glycol) (PEG) microspheres were designed and synthesized by polymerization of Dex-GMA emulsified in an aqueous PEG solution. Dex-GMA was prepared by substituting the hydroxyl groups in Dex by GMA. The drug loading and in vitro drug release was evaluated by routine procedure and the biological activity of BMP-loaded microspheres was studied by experimental cytology methods. Recombinant human bone morphogenetic protein-2 (rhBMP-2) were entrapped in dextran-derived microspheres quantitatively and with full preservation of their biological activity. In vitro release kinetics indicated that dextran-derived microspheres could retain rhBMP-2 in a variable manner depending on the preparation and degradation of the microspheres. The release profiles of rhBMP-2 from microspheres as a function of time showed that rhBMP-2 releasing kinetics in vitro fitted to first-order and Higuchi equations. The release profile in vitro was in accord with two phases kinetics law and more than 60% drug were released during 20 days. Cytology studies showed rhBMP-2 microspheres have good biological effects on cultured periodontal ligament cells, and could achieve a longer action time than concentration of rhBMP-2 solution. These properties make those microspheres interesting osteo-conductive BMP carriers, allowing to decrease the amount of implanted factor required for tissue regeneration. PMID- 16260106 TI - Influence of electricity on post-mortal body temperature. AB - In a case of suicidal application of electricity differences between the rectal temperature of the body and the suspected time of death were observed. In order to answer the question whether an electric current from hand to hand over >30 min led to a rise in body temperature FEM-based computer simulations and animal experiments were carried out. Both resulted in a warming of the soft parts in the arm without warming the body core. Thus a temperature-based estimation of the time since death can also be used in cases with electricity as the cause of death. Besides, in the animal experiment we found a spontaneous rise in the body core temperature even without application of electricity which may be a reason for the typical temperature plateau after death. PMID- 16260107 TI - Population data for 11 Y-chromosome STRs in northeast China Han. AB - Allele and haplotype frequencies for the 12 Y-specific short tandem repeats (STR) loci DYS19, DYS385a/b, DYS389I/II, DYS390, DYS391, DYS392, DYS393, DYS437, DYS438, DYS439 (PowerPlex Y System STR Amplification Kit, Promega) were determined in a population sample of 187 unrelated China Han in northeast China. PMID- 16260108 TI - [Neuraminidase inhibitors in the general practice management of influenza: who prescribe them, when and with which results?]. AB - OBJECTIVE: To describe in real-life conditions the flu therapeutic management, motivations to prescribe or not NAI (General Practitioners' (GPs) characteristics, decisional factors) and treated patients' course. DESIGN: A prospective, longitudinal, pharmacoepidemiological study involved 305 GPs in France during 2002-2003 winter epidemic peak. All patients>or=1 year old, with a clinical diagnostic of flu were included. RESULTS: One hundred and eighty-five GPs (150 NAI prescribing and 30 non-prescribing physicians) have included at least 1 patient. Prescribing physicians were the best informed on flu and NAI. 660 patients were analysed (250 NAI+ and 410 NAI-). 66% of NAI+ and 40% of NAI- attended to a consultation within 24 h (P<0.001). 31% of NAI+ and 20% of NAI- had a visit at home (P=0.002). Among the patients without complication at inclusion (N=585), 3% of NAI+ received an antibiotherapy vs 13% of NAI- (P<0.001). 43% of the patients had a sick leave, shorter for the NAI+ than NAI- (respectively, 3.7+/-1.7 vs 4.2+/-1.7 days, p=0.017). NAI was taken within 3 hours (median) after prescription by the 78% of the patients who returned their diary cards. The NAI+ patients had a faster improvement of symptoms than NAI- (within 24 h, respectively: 18 vs 5%, P<0.001) and they returned faster to routine activities (within 48 h, respectively: 27 vs 11%, P<0.001). CONCLUSIONS: This study evidenced the good use of NAI by the physicians. It confirms their therapeutic efficacy in real-life conditions and suggests their prescription allows decreasing antibiotic co-prescriptions and sick leaves duration, profits to consider in NAI benefit/risk ratio. PMID- 16260109 TI - Aging affects but does not eliminate the enzymatic antioxidative response to hypoxia/reoxygenation in cerebral cortex. AB - The effect of aging on basal and hypoxia/reoxygenation levels of both oxidative stress (protein carbonyl and TBARS) and antioxidative-enzyme activity (Cu/Zn-SOD; Mn-SOD; Catalase, CAT; Se-independent and Se-dependent glutathione peroxidase, GPX; glutathione transferase, GST and glutathione reductase, GR) has been studied in the cerebral cortex of adult and old rats. Oxidative stress markers increased with aging and show an age-dependent post-hypoxic response. Moreover, aging caused either no change (GST, GR and CAT) or an increase (Se-GPX, Cu/Zn-SOD, Mn SOD) in the basal activity of the enzymes analysed. Only Se-independent GPX activity decreases. However, we detected an age-dependent response of SODs to the hypoxic injury. The early and sustained Cu/Zn-SOD activity rise in adult animals became late and weak in aged animals. Meanwhile, aging slowed the Mn-SOD post hypoxic response although this activity was consistently higher in aged rats. Aging eliminated the post-hypoxic CAT response, but, perhaps offset by increased GPX activity, did not affect the GST response and slightly reduced post-hypoxic GR activity. In conclusion, aging rise basal ROS production, does not diminish or even increase the antioxidative-enzyme activity, and may slow but does not usually eliminate the enzymatic antioxidant response to the increased post hypoxic ROS generation. PMID- 16260110 TI - Structure elucidation and conformational study of V8: a novel synthetic non peptide AT(1) antagonist. AB - AT(1) antagonists constitute the most recent class of antihypertensive drugs which act through the Renin Angiotensin System (RAS). In an effort to comprehend their stereoelectronic features, a study was initiated to compare the conformational properties of drugs already marketed for the treatment of hypertension with synthetic ones, possessing common structural characteristics. In this study, the synthetic AT(1) antagonist V8 is structurally elucidated and its conformational properties are studied through a combination of NMR spectroscopy and computational analysis. Its conformational properties are compared with those of the structurally similar prototype AT(1) antagonist losartan. PMID- 16260111 TI - Imipenem-resistant Achromobacter xylosoxidans carrying blaVIM-2-containing class 1 integron. AB - We characterized seven isolates of imipenem-resistant Achromobacter xylosoxidans that were isolated from patients hospitalized in the intensive care unit at a tertiary hospital in Korea during 2001 to 2003. From the analysis with an isoelectric focusing, polymerase chain reaction, and sequencing methods, all isolates were found to produce VIM-2, OXA-30, and chromosomal AmpC beta-lactamase with a pI of 8.4. They showed a similar antibiogram, which were resistant to all tested aminoglycosides as well as beta-lactams including imipenem (16-32 mg/L) and aztreonam (128 mg/L), and a same DNA fingerprinting pattern by random amplified polymorphic DNA analysis, suggesting that these originated from a single clone. From the analysis of integron structure carried by an isolate of A. xylosoxidans CBU1760, bla(VIM-2) was found to be part of a gene cassette carried on a class 1 integron (3.4 kb) containing three aacA4 gene cassettes. This is the first report of bla(VIM-2) in A. xylosoxidans. PMID- 16260112 TI - Steroid hormone receptor expression in male breast cancer. AB - AIMS: To investigate expression of the steroid hormone receptors estrogen receptor (ER)-alpha and -beta, progesterone receptor (PR) and androgen receptor (AR) in male breast cancer. METHODS: Specimens from 16 male breast cancers were immunostained for ERalpha, ERbeta, PR and AR. FINDINGS: Eighty-seven percent of tumours expressed ERalpha, 93% PR, 87% ERbeta and 87% AR. Staining for ERalpha and PR was confined exclusively to the nuclei of epithelial cells with some heterogeneity. Nuclear immunoreactivity was also observed with AR. Again this was restricted to epithelial cells but tended to be more uniform. ERbeta was seen in the nuclei of epithelial cells and also in stromal fibroblasts and lymphocytes. Analysis of serial sections revealed a similar pattern of staining with ERbeta and AR in epithelial cells. CONCLUSIONS: In addition to expression of the better known steroid receptors, ERalpha, PR and AR, we have demonstrated a high rate of expression of ERbeta in male breast cancer. This is in keeping with the generally high steroid receptor expression seen in males. However, the abundance of ERbeta expressed in this small series of male breast cancer is in contrast to female breast cancer where ERbeta expression is often reduced. PMID- 16260114 TI - Insecta immune-cognitive interactions. PMID- 16260113 TI - Application of boron-entrapped stealth liposomes to inhibition of growth of tumour cells in the in vivo boron neutron-capture therapy model. AB - Tumour cell destruction in boron neutron-capture therapy (BNCT) is due to the nuclear reaction between (10)B and thermal neutrons. It is necessary for effective BNCT therapy to accumulate (10)B atoms in the tumour cells. The delivery system consisted of polyethylene-glycol (PEG) binding liposomes (DPPC/cholesterol/DSPC-PEG2000) with an entrapped (10)B-compound and we evaluated the cytotoxic effects of intravenously injected (10)B-PEG-liposomes on human pancreatic carcinoma xenografts in nude mice with thermal neutron irradiation. After thermal neutron irradiation of mice injected with (10)B-PEG-liposomes, growth of AsPC-1 tumours was suppressed relative to controls. Injection of (10)B PEG-liposomes caused the greatest tumour suppression with thermal neutron irradiation in vivo. These results suggest that intravenous injection of (10)B PEG-liposomes can increase the retention of (10)B atoms by tumour cells, causing suppression of tumour growth in vivo, after thermal neutron irradiation. PMID- 16260115 TI - Apoptotic effects of 25-hydroxycholesterol in immature rat Sertoli cells: prevention by 17beta-estradiol. AB - The aim of the present study was to determine whether or not apoptosis occurs in Sertoli cells in presence of 25-hydroxycholesterol, an oxysterol derived from cholesterol-containing foods or endogenous oxidation. Here, we provide evidence that 25-hydroxycholesterol can induce cultured Sertoli cells of immature rat to undergo apoptosis. The cell death was identified by analysis of fragmented DNA detected using enzyme-immunoassay. After 48 h of treatment with 50 microM of 25 hydroxycholesterol, apoptosis increased by 70% in Sertoli cells. Moreover, 50 microM of 25-hydroxycholesterol inhibited the incorporation of [14C] acetate into cholesterol by 70%. Addition of mevanolate to prevent isoprenoid deficiency do not inhibit the apoptosis generated by 25-hydroxycholesterol. In contrast, this increase of DNA fragmentation was reversed by addition of caspase-3 inhibitors as Ac-DEVD-CHO or Ac-ESMD-CHO. Bcl-2 mRNA level in the Sertoli cells decreased by 60% after 24 h exposure to 25-hydroxycholesterol. In parallel, Bax mRNA level increased by 40% in the Sertoli cells incubated in presence of 50 microM of 25 hydroxycholesterol. Physiological concentrations of 17beta-estradiol (10 or 100 nM) elicited a significant protection on apoptosis generated by 25 hydroxycholesterol in Sertoli cells. Our results show that the 25 hydroxycholesterol would control the cholesterol synthesis without toxic effect in immature rat Sertoli cells, these cells being able to protect themselves by estradiol production. PMID- 16260116 TI - Methods for reducing interference in the Complementary Learning Systems model: oscillating inhibition and autonomous memory rehearsal. AB - The stability-plasticity problem (i.e. how the brain incorporates new information into its model of the world, while at the same time preserving existing knowledge) has been at the forefront of computational memory research for several decades. In this paper, we critically evaluate how well the Complementary Learning Systems theory of hippocampo-cortical interactions addresses the stability-plasticity problem. We identify two major challenges for the model: Finding a learning algorithm for cortex and hippocampus that enacts selective strengthening of weak memories, and selective punishment of competing memories; and preventing catastrophic forgetting in the case of non-stationary environments (i.e. when items are temporarily removed from the training set). We then discuss potential solutions to these problems: First, we describe a recently developed learning algorithm that leverages neural oscillations to find weak parts of memories (so they can be strengthened) and strong competitors (so they can be punished), and we show how this algorithm outperforms other learning algorithms (CPCA Hebbian learning and Leabra at memorizing overlapping patterns. Second, we describe how autonomous re-activation of memories (separately in cortex and hippocampus) during REM sleep, coupled with the oscillating learning algorithm, can reduce the rate of forgetting of input patterns that are no longer present in the environment. We then present a simple demonstration of how this process can prevent catastrophic interference in an AB-AC learning paradigm. PMID- 16260117 TI - Genetics of autoimmune myasthenia gravis: the multifaceted contribution of the HLA complex. AB - The HLA complex plays a prominent role in predisposition to many autoimmune diseases. Thus far, the highly polymorphic class I and class II loci have been considered as the prime candidates to explain this role. There is nonetheless growing evidence that other closely linked HLA loci are also involved in autoimmune susceptibility. Their search, however, has been hampered by the often strong linkage disequilibria, i.e. the non-random association of alleles at linked loci, across the HLA complex. Here, we discuss recent work from our laboratory on the dissection of this emblematic genetic region in a model autoimmune disease, acquired myasthenia gravis (MG). PMID- 16260118 TI - Anti-TNF therapy: where have we got to in 2005? AB - The blockade of TNF has had significant impact on the therapy of a number of chronic autoimmune diseases. In this chapter we review the concepts leading up to this therapy in rheumatoid arthritis (RA), how it spreads into other autoimmune diseases, and how greater understanding of its use has led to augmented therapeutic benefit. There are still many limitations, but the prospects for the future are intriguing. PMID- 16260119 TI - Signalling via cGMP: lessons from Drosophila. AB - Guanosine 3', 5'-cyclic monophosphate (cGMP) signalling has received increasing attention over the last decade, since the discovery of the gaseous signalling molecule, nitric oxide, which activates cGMP synthesis. Furthermore, research into cGMP signalling has also been stimulated by the development of Viagra and pharmacologically active related compounds, which act to prevent cGMP breakdown. While much is known about the biochemical aspects of components of the cGMP signalling pathway, the precise in vivo roles of such components have only recently come to light through work in model organisms. This review outlines recent work utilising the genetic model organism Drosophila melanogaster in studies of organotypic cGMP signalling. While organisms such as Drosophila may not be the obvious choice for such studies, use of this model has proved that unique and detailed insights for cGMP signalling can be achieved. PMID- 16260120 TI - Specific cognitive deficits and differential domains of social functioning impairment in schizophrenia. AB - There is considerable inconsistency in findings regarding the relationship between specific cognitive deficits and social impairment in patients with schizophrenia. This inconsistency may relate to variability across studies in how social functioning is measured and preliminary evidence suggests that different indices of social functioning (e.g., laboratory test, community assessment) may have different cognitive correlates. The present study examined this issue by evaluating the relationships between cognitive deficits (including social cognitive deficits), role-play test performance, and community social functioning in 28 inpatients with schizophrenia. We expected the two measures of social functioning to have only modest convergence with each other. Moreover, informed by the literature on cognitive functioning in schizophrenia, we identified specific cognitive processes that were hypothesized to be associated with role play performance (delayed verbal memory and attentional vigilance) and social functioning in the community (delayed verbal memory and executive functioning). As expected, the two measures of social functioning were modestly correlated with each other. Community social functioning was associated with a relatively constrained pattern of cognitive deficits and received a significant contribution (Deltar2=0.24) from specific cognitive processes beyond that of general cognitive functioning and symptom severity. In contrast to our hypotheses, role-play test performance was associated with a wide range of cognitive impairments and received little contribution from the specific cognitive processes beyond the effects of general cognitive functioning. Community social functioning, but not role-play test performance, was significantly associated with social cognition. These findings highlight the importance of conceptualizing social functioning as a multidimensional construct for schizophrenia research. PMID- 16260122 TI - NMR as a tool in the investigation of fundamental problems in ordered liquids. AB - An overview is presented of modern NMR techniques and a variety of experimental and theoretical tools employed in the study of solutes dissolved in liquid crystals. The NMR techniques involve multiple quantum and spectral subtraction methods. In addition, various experimental and theoretical tools are discussed, including: the theoretical background of observed order parameters; the use of 'magic mixtures' to separate orientational contributions; the reorientation vibration interaction; the use of model calculations based on size and shape of the various solutes; and the use of computer simulations. Applications to the benchmark probe molecules hydrogen, methane, ethane, and butane and their isotopomers are treated. PMID- 16260121 TI - Effect of physicochemical properties of cyclic terpenes on their ex vivo skin absorption and elimination kinetics. AB - BACKGROUND: The terpenes disturb lipid arrangement in the intercellular region of the stratum corneum (SC) that leads to the increased permeability of the skin. This effect is used in technology of transdermal drug forms and depends on physicochemical properties of terpenes and their amounts penetrated to the stratum corneum; however terpenes do not need penetrate into viable skin tissue and this event is not even desired. OBJECTIVE: To correlate skin absorption and elimination kinetics of four cyclic terpenes, namely alpha-pinene, beta-pinene, eucalyptol and terpinen-4-ol, applied as neat substance with their physicochemical properties. METHODS: The terpenes were applied onto the human skin in vitro, and after 1-4 h their content in the separated by a tape-stripping method stratum corneum layers and in the epidermis/dermis was determined using GC. Similarly, the amounts of terpenes in the skin were analysed during 4 h following 1 h absorption. RESULTS: The fastest and progressive penetration into all skin layers was observed for terpinen-4-ol. All studied terpenes are absorbed in the viable epidermis/dermis, however penetration into this layers is time dependent process, constantly increasing during 4 h. Like for stratum corneum, the largest cumulation in epidermis/dermis was observed for terpinen-4-ol. The elimination of terpenes from the stratum corneum was fast, especially in deeper layers, and much faster if the initial cumulation was small. CONCLUSION: Investigated cyclic terpenes represent different penetration and elimination characteristics and do not permeate across the skin to the acceptor medium due to large cumulation in the skin tissue. The penetration of terpenes into stratum corneum is greater if their log P-value is close to 3. PMID- 16260123 TI - Cross-modal plasticity in deaf subjects dependent on the extent of hearing loss. AB - Cross-modal plasticity in deaf subjects is still discussed controversial. We tried to figure out whether the plasticity is dependent on the extent of hearing loss. Three groups of volunteers, comprising twelve individuals each, were investigated. They were characterized by three distinctive features, one had normal hearing, the other one lost hearing and the third had only minimal residual hearing ability. All participants, except those of group one, were capable of using German Sign Language (GSL). The groups were studied with functional MRI in a standard block design during individuals' watching sign language videos alternating with black frame. During sign language conditions, deaf subjects revealed a significant activation of the auditory cortex in both hemispheres comprising Brodmann areas (BA) 42 and 22 corresponding to the secondary associative auditory areas. Additionally, activation of the angular and supramarginal gyrus was seen. Activation of the primary auditory cortex was revealed in deaf subjects with total hearing loss during sign language tasks but not in subjects with residual hearing ability. In conclusion our results indicate a cortical reorganization of the auditory cortex comprising primary auditory fields only present in subjects with total hearing loss. PMID- 16260124 TI - Arithmetic ability and parietal alterations: a diffusion tensor imaging study in velocardiofacial syndrome. AB - Velocardiofacial syndrome (VCFS) is a congenital anomaly that causes somatic as well as cognitive and psychiatric impairments. Previous studies have found specific deficits in arithmetic abilities in subjects with VCFS. In this study, we investigated whether abnormalities in white matter pathways are correlated with reduced arithmetic ability. Nineteen individuals with VCFS aged 7-19 years received diffusion-weighted magnetic resonance imaging (MRI) scans. A linear regression model was used to correlate fractional anisotropy (FA) values with scores of the arithmetic subscale on the WISC/WAIS on a voxel-by-voxel basis, after covarying for any IQ- and age-related effects. There was a statistically significant positive correlation between the arithmetic score on the WISC/WAIS and FA values in white matter tracts adjacent to the left supramarginal and angular gyri, as well as along the left intraparietal sulcus. Inferior parietal lobe white matter structural aberrations may contribute to reduced arithmetic ability in VCFS. PMID- 16260125 TI - Synthesis of novel phenylacetic acid derivatives with halogenated benzyl subunit and evaluation as aldose reductase inhibitors. AB - In the course of our ongoing studies several substituted benzyloxyphenylacetic acids were prepared. Comparison of their aldose reductase inhibition with the biological activity obtained for recently evaluated benzoic acid analogues revealed the critical role of a methylene spacer between the aromatic core and the acidic function. Starting from the most potent derivative (i.e. 5d, IC50 = 20.9 microM) further structural modifications were performed and their influence on the inhibitory effect was established. PMID- 16260128 TI - Chemical biotechnology: what's new, what's next? PMID- 16260129 TI - Fear extinction in rodents: basic insight to clinical promise. AB - Fear extinction, the reduction of fear by repeated exposure to the object of fear, is a crucial paradigm of inhibitory learning and the acknowledged preclinical model for behavior therapy of human anxiety. Recent insights have clarified roles for infralimbic prefrontal cortex, hippocampus and periaqueductal gray in extinction learning, while maintaining a central role for the basolateral amygdaloid nucleus in the acquisition and storage of this learning. Simultaneously, molecular insights have implicated several neurotransmitter and second messenger systems in extinction learning, and revealed that extinction is surprisingly easy to improve, yielding the promise of a novel approach to improved psychiatric treatments for a variety of human anxiety disorders. PMID- 16260130 TI - How social experiences influence the brain. AB - Social experiences throughout life influence gene expression and behavior, however, early in development these influences have a particularly profound effect. In mammals, mother-infant interactions are the primary source of social stimulation and result in long-term changes in offspring phenotype. This has previously been demonstrated in rodents and primates, however, recent studies in rats have advanced our understanding of how these influences are achieved at a mechanistic level, through epigenetic modification, and provide a model for studying the transmission of social behavior across generations. These studies emphasize the importance of a life-history approach to the study of brain development; incorporating information about genetic background, prenatal and postnatal maternal care received, and post-weaning social interactions of an individual, in addition to the social environment experienced by previous generations. PMID- 16260126 TI - Modified conventional hard gelatin capsules as fast disintegrating dosage form in the oral cavity. AB - Fast disintegrating capsules for administration in the oral cavity were prepared either by perforation or by vacuum-drying of conventional hard capsules. When compared to other fast disintegrating dosage forms (e.g. lyophilized sponges or tablets), these capsules have various advantages, in particular, a high drug loading capacity and no compression steps. The disintegration time of conventional hard gelatin capsules (HGC) was reduced from 91 to 39 s by introducing 6-10 small holes (diameter =25-50 microm) into the capsule shell. Vacuum-drying of conventional hard gelatin capsules resulted in brittle capsules, which broke rapidly in the oral cavity. The brittleness of the hard gelatin capsules correlated well with their moisture content. The critical moisture value for sufficient brittleness of hard gelatin capsules was <4% w/w. In contrast, HPMC capsules remained flexible, even at low moisture content. The moisture uptake of various capsule fillers was in the order of Avicel PH101 > lactose > Avicel PH112 > or = mannitol. Hard gelatin capsules filled with mannitol and packaged in bottles with silica gel kept their desired brittleness during 6 months storage at various relative humidities. PMID- 16260127 TI - Serum lipid fatty acids, phonological processing, and reading in children with oral clefts. AB - Reading skill is suggested to be related to phonological processing ability and polyunsaturated fatty acids (PUFAs). Here we investigated whether fatty acids (FAs) are related to phonological processing, whether the relations between PUFAs and reading generalize to other FAs, whether these relations are mediated by phonological processing, and whether relations of FAs are specific for language related functions. Blood samples of 49 ten-year-old children with oral clefts were collected for FA proportion analysis in serum cholesteryl esters and phospholipids. On the same day, they performed tasks of phonological processing, reading, and both verbal and nonverbal intelligence. Sequential regression analyses (adjusted for age, gender, and cleft type) showed that phonological processing was inversely related to myristic acid in phospholipids and positively related to eicosapentaenoic acid in cholesteryl esters. Reading was inversely related to palmitoleic and gammalinolenic acids in phospholipids. The relations between FAs and reading were not mediated by phonological processing and FAs related only to language-related functions. PMID- 16260131 TI - Tetrahydrobenzothiophene inhibitors of hepatitis C virus NS5B polymerase. AB - A novel series of selective HCV NS5B RNA dependent RNA polymerase inhibitors has been disclosed. These compounds contain an appropriately substituted tetrahydrobenzothiophene scaffold. This communication will detail the SAR and activities of this series. PMID- 16260132 TI - Synthesis of N-benzylated-2-aminoquinolines as ligands for the Tec SH3 domain. AB - In recent work, we have been developing 2-aminoquinolines as ligands for Src Homology 3 (SH3) domains, so far the only reported examples of small-molecule ligands for these domains. In this paper, we report the synthesis of a series of N-benzylated-2-aminoquinolines by reductive amination of aryl aldehydes with 2 aminoquinoline. These ligands bound the SH3 domain with ca. one and a half to twofold reduced affinity relative to 2-aminoquinoline; however, some evidence was found to suggest that the benzylic substituents made new contacts with the SH3 domain surface. These results provide useful SAR information that may assist in future ligand design. PMID- 16260133 TI - The identification of potent and selective imidazole-based inhibitors of B-Raf kinase. AB - A novel triarylimidazole derivative, SB-590885 (33), bearing a 2,3-dihydro-1H inden-1-one oxime substituent has been identified as a potent and extremely selective inhibitor of B-Raf kinase. PMID- 16260134 TI - Hydroxylated analogues of the orally active broad spectrum antifungal, Sch 51048 (1), and the discovery of posaconazole [Sch 56592; 2 or (S,S)-5]. AB - As part of a detailed study, the syntheses, biological activities, and pharmacokinetic properties of hydroxylated analogues of the previously described broad spectrum antifungal agents, Sch 51048 (1), Sch 50001 (3), and Sch 50002 (4), are described. Based on an overall superior profile, one of the alcohols, Sch 56592 (2), was selected for clinical studies. PMID- 16260135 TI - Biomimetic synthesis and ultrastructural characterization of a zerovalent gold hydroxyapatite composite. AB - This manuscript describes one-pot, in situ synthesis of organoapatite-gold nanoparticle composite and its characterization by XRD, energy dispersive X-ray analysis, atomic force, and scanning electron microscopy. This methodology offers a different approach into the synthesis of zerovalent metal-mineral-phase nanocomposites having potential application as osteointegrative ceramics. PMID- 16260136 TI - Tip of another iceberg: Drosophila serpins. AB - Serpins are serine protease inhibitors with a conserved structure that have been identified in nearly all species and act as suicide substrates by binding covalently to their target proteases. Serpins regulate various physiological processes and defence mechanisms. In humans, several serpin mutations are linked to diseases. The genome of Drosophila melanogaster encodes 29 serpins and even more serine proteases. To date, three serpins have been investigated in detail. Spn27A controls the Toll pathway during early development and is involved in defence reactions in adult flies. SPN42DaA is an inhibitor of furin, a subtilisin like convertase that is required for pro-protein maturation. Spn43Ac controls the Toll pathway during the immune response. In each case, Drosophila genetics has shed new light on the function of these serine protease inhibitors. PMID- 16260137 TI - Cytotoxic T lymphocyte exocytosis: bring on the SNAREs! AB - Despite our general understanding of membrane traffic, the molecular machinery at the immunological synapse (IS) that regulates exocytosis of lytic granules from cytotoxic T lymphocytes (CTLs) remains elusive. The identification of disease causing mutations in the small GTPase Rab27a, priming factor Munc13-4 and fusion protein syntaxin11 has defined an important role for these proteins in CTL exocytosis. In addition, the demonstration of a direct interaction in vitro between Rab27a and Munc13-4 suggests the possibility that the Rab27a-Munc13-4 cascade might regulate CTL exocytosis by engaging SNAREs such as syntaxin11. We propose that these SNAREs are likely to mediate the fusion of lytic granules with the plasma membrane of the IS. PMID- 16260138 TI - The experience of agency nurses working in a London teaching hospital. AB - This Husserlian phenomenological study explored the lived experience of eight full-time agency nurses working in the environment of intensive care in London, United Kingdom. In-depth interviews were used to ascertain the participants' experiences of full-time agency nursing in intensive care. Colaizzi's [Colaizzi P. Psychological research as the phenomenologist views it. In: Vale R, King M, editors. Existential-phenomenological alternatives for psychology. London: Oxford University Press; 1978, p. 48-71] method of data analysis was utilised. Thematic analysis identified three cluster themes: the shared experience of lacking confidence; the shared experience of deskilling; and the shared experience of feelings of isolation. The theme of the shared experience of lacking confidence identified the importance of support and understanding of agency nurses' needs in the ICU. The theme of the shared experience of deskilling illuminated participants concern at not having the opportunity to care for patients of higher acuity. The theme of the shared experience of feelings of isolation highlighted that often participants did not feel they "belonged to a team". Study findings suggest the need to include: the implementation of protocols within the clinical setting; the provision of performance feedback to agency nurses; and the consideration of ongoing structured professional development and education for ICU agency nurses. PMID- 16260139 TI - The saving lives and 100,000 Lives programmes: good news for critical care nurses. PMID- 16260140 TI - Effect of skin movement artifact on knee kinematics during gait and cutting motions measured in vivo. AB - Eight healthy male subjects had intra-cortical bone-pins inserted into the proximal tibia and distal femur. Three reflective markers were attached to each bone-pin and four reflective markers were mounted on the skin of the tibia and thigh, respectively. Roentgen-stereophotogrammetric analysis (RSA) was used to determine the anatomical reference frame of the tibia and femur. Knee joint motion was recorded during walking and cutting using infrared cameras sampling at 120Hz. The kinematics derived from the bone-pin markers were compared with that of the skin-markers. Average rotational errors of up to 4.4 degrees and 13.1 degrees and translational errors of up to 13.0 and 16.1mm were noted for the walk and cut, respectively. Although skin-marker derived kinematics could provide repeatable results this was not representative of the motion of the underlying bones. A standard error of measurement is proposed for the reporting of 3D knee joint kinematics. PMID- 16260141 TI - The kinematic and kinetic effects of solid, hinged, and no ankle-foot orthoses on stair locomotion in healthy adults. AB - This study compared the effects of a unilateral solid ankle-foot orthosis (AFO), hinged AFO and no AFO (shoe) worn by healthy adults on pelvic angles, lower extremity joint angles, moments and powers, and temporal-spatial gait characteristics during stair locomotion. A convenience sample of 19 healthy adults participated in this repeated measures design with subjects serving as their own controls. Subjects ambulated on stairs wearing a left shoe and either a right solid AFO, hinged AFO or shoe. Kinematic and kinetic data were collected with motion analysis equipment and a force plate for the three conditions. Pelvic angles and right hip, knee and ankle angles, moments and powers during stance were compared to determine differences among the conditions. Subjects wearing either orthosis walked slower during stair locomotion and with a shorter right single limb support time during descent. Sagittal knee and ankle angles, moments and powers were similar in individuals wearing a hinged AFO or shoe during pull up (PU) in ascent and controlled lowering (CL) in descent. Decreased ankle dorsiflexion angle, plantar flexion power, knee flexion angle and extensor moment were seen in subjects wearing a solid AFO as compared to a hinged AFO during PU in ascent and CL in descent. Findings contributed to understanding how biomechanical changes imposed at the ankle by a unilateral solid AFO resulted in more kinetic and kinematic compensations than the hinged AFO in healthy adults without the confounding effects of neuromuscular impairments. PMID- 16260142 TI - Accuracy and repeatability of the Pedar Mobile system in long-term vertical force measurements. AB - Portable insole pressure systems can be used to measure the vertical force during long-term (hours) measurements to determine the patient's amount of weight bearing during daily activities in the hospital and at home. Especially for long term measurements, the amount and duration of loading pressure insoles can have a large influence on the accuracy, as previous studies found a time-dependent behavior after a relatively short period (minutes) of constant loading. Therefore, this study assessed the accuracy and repeatability of a portable capacitive insole system (Pedar, Novel(GmbH)) to measure vertical force during long-term loading. Static loading experiments were performed during which the Pedar insoles were loaded with 5 and 10 N/cm2 for 7 h. Dynamic loading experiments were performed with one Pedar insole which was cyclically loaded with 300, 500 and 1000 N during two sessions of 1200 load cycles. The static and dynamic experiments were repeated 3 days later. Accuracy, due to offset drift, decreased in time during the start of the static experiments (percent error: 1.9% to 0.3% at hour 0; 26.3% to 34% at hour 7). The percent error for the dynamic experiments ranged from -16% to -19%, from -3% to -7% and from -8% to approximately 0% when the insole was loaded with 300, 500 and 1000 N, respectively. The amount of drift ranged from 12 to 62 N for the 500 and 1000 N loads, respectively. The mean day-to-day percentage difference for the static and dynamic experiments ranged from -2.3% to 0.5%, and from -2.9% to 3.0%, respectively. The results indicate that drift correction is necessary for accurate assessment of vertical force by the Pedar Mobile system to determine the amount of weight bearing during long-term measurements. PMID- 16260143 TI - Phospholipase C-gamma: diverse roles in receptor-mediated calcium signaling. AB - Ca2+ is a universal signal: the dynamic changes in its release and entry trigger a plethora of cellular responses. Central to this schema are members of the phospholipase C (PLC) superfamily, which relay information from the activated receptor to downstream signal cascades by production of second-messenger molecules. Recent studies reveal that, in addition to its enzymatic activity, PLC gamma regulates Ca2+ entry via the formation of an intermolecular lipid-binding domain with canonical transient receptor potential 3 (TRPC3) ion channels. This complex, in turn, controls TRPC3 trafficking and cell-surface expression. Thus, TRPC3 ion channels are functionally linked to both lipase-dependent and independent activities of PLC-gamma. Understanding the underlying molecular mechanisms that regulate this complex will probably clarify the processes of receptor-activated Ca2+ entry. PMID- 16260144 TI - Amino acid specificity in translation. AB - Recent structural and biochemical experiments indicate that bacterial elongation factor Tu and the ribosomal A-site show specificity for both the amino acid and the tRNA portions of their aminoacyl-tRNA (aa-tRNA) substrates. These data are inconsistent with the traditional view that tRNAs are generic adaptors in translation. We hypothesize that each tRNA sequence has co-evolved with its cognate amino acid, such that all aa-tRNAs are translated uniformly. PMID- 16260145 TI - The mitochondrial permeability transition pore and the Ca2+-activated K+ channel contribute to the cardioprotection conferred by tumor necrosis factor-alpha. AB - Pretreatment with tumor necrosis factor-alpha (TNF-alpha) is known to trigger cardioprotection and it can activate multiple downstream signaling cascades. However, it is not known whether the mitochondrial permeability transition pore and the Ca(2+)-activated K(+) channel (K(Ca) channel) are involved in the TNF alpha-induced cardioprotection. In the present study, we examined whether TNF alpha inhibits pore opening and activates the K(Ca) channel in the cardioprotection. In isolated rat hearts subjected to 30 min of regional ischemia and 120 min of reperfusion, pretreatment with 10 U/ml TNF-alpha for 7 min followed by 10 min washout improved the recovery of rate-pressure product (RPP=left ventricular developed pressure x heart rate) and coronary flow (CF) during reperfusion, and reduced the infarct size and release of lactate dehydrogenase (LDH). Administration of 20 micromol/L atractyloside, a pore opener, for the last 5 min of ischemia and first 15 min of reperfusion, and pretreatment with 1 micromol/L paxilline, an inhibitor of the K(Ca) channel, for 5 min before ischemia, attenuated the recovery of RPP and CF, and the reductions of infarct size and release of LDH induced by TNF-alpha. On the other hand, administration of 10 micromol/L NS 1619, an opener of the K(Ca) channel, for 10 min before ischemia, decreased the infarct size and LDH release, and improved contractile functions and CF; these effects were attenuated by atractyloside. Pretreatment with 0.2 micromol/L cyclosporin A for the last 5 min of ischemia and first 15 min of reperfusion showed similar effects to those of TNF-alpha, and they were not attenuated by paxilline. In mitochondria isolated from hearts pretreated with 10 U/ml TNF-alpha for 7 min, a significant inhibition of Ca(2+) induced swelling was observed. Furthermore, paxilline attenuated the inhibition of Ca(2+)-induced mitochondrial swelling by TNF-alpha. These findings indicate that TNF-alpha protects the myocardium against ischemia and reperfusion injury by inhibiting mitochondrial permeability transition pore opening as well as activating K(Ca) channels, probably the mitochondrial K(Ca) channel, which is upstream from the pore. PMID- 16260146 TI - Stabilization of ubiquitous mitochondrial creatine kinase preprotein by APP family proteins. AB - Amyloid precursor protein (APP) is involved in the pathogenesis of Alzheimer's disease (AD). However, the physiological role of APP and its family members is still unclear. To gain insights into APP function, we used a proteomic approach to identify APP interacting proteins. We report here for the first time a direct interaction between the C-terminal region of APP family proteins and ubiquitous mitochondrial creatine kinase (uMtCK). This interaction was confirmed in vitro as well as in cultured cells and in brain. Interestingly, expression of full-length and C-terminal domain of APP family proteins stabilized uMtCK preprotein in cultured cells. Our data suggest that APP may regulate cellular energy levels and mitochondrial function via a direct interaction and stabilization of uMtCK. PMID- 16260147 TI - Validation of ELISA for the determination of anti-ricin immunoglobulin G concentration in mouse sera. AB - An enzyme-linked immunosorbent assay (ELISA) for the determination of anti-ricin immunoglobulin G (IgG) concentration in mouse sera was systematically validated. The results obtained throughout the validation process strongly demonstrated that the ELISA was reliable, reproducible, and suitable for its intended use. The assay had a high level of precision within and between runs, was specific for the anti-ricin IgG, and showed no interference with a number of different serum matrices. The assay exhibited excellent accuracy, linearity, and stability. The mean recovery of four test samples with different known concentrations was 100.9+/-11.3%, 102.7+/-10.8%, 99.0+/-7.2%, and 95.9+/-11.3%, respectively (n=10). The mean recovery of the observed anti-ricin IgG concentration of three quality control samples run on 73 plates to their nominal concentrations was 100.1+/ 7.3%, 100.2+/-5.8%, and 103.7+/-8.1%; and the coefficient of variation (CV) was 7.3%, 5.8%, and 7.8%, respectively. The back-calculated anti-ricin IgG concentration, %CV, and relative error of seven standards from the calibration curves run in the entire validation study were analyzed (n=7 x 73). The results indicated that the four-parameter logistic (4PL) equation, y=(a-d)/(1+(x/c)b)+d, provided an accurate representation of a sigmoidal relationship between the measured response and the logarithm of observed concentration of anti-ricin IgG in mouse sera for this ELISA. The lower limit of quantification and upper limit of quantification of the calibration curve were 3.3 ng/ml and 82.8 ng/ml, respectively. The measurable range of the assay would cover all possible anti ricin IgG concentrations in mouse sera stimulated with a ricin vaccine candidate, when the test sera are measured at a 1:800 starting dilution followed by four additional fourfold serial dilutions. PMID- 16260148 TI - Construction, expression, and purification of HIV-TAT-survivin (T34A) mutant: a pro-apoptosis protein in Escherichia coli. AB - As a novel member of the IAP family, survivin was observed to express in the most common human cancers. Anti-cancer therapy targeting survivin has drawn considerable attention. This report presented firstly construction of recombinant plasmid pRSET-B-TAT-survivin (T34A), expression in Escherichia coli, purification, renaturation, and bioactivity. The cDNA encoding survivin was cloned by RT-PCR from breast cancer cell lines B-cap37. Expression vector pRSET-B TAT-survivin (T34A) was constructed by PCR after survivin was mutated by PCR site directed mutagenesis. Recombinant TAT-survivin (T34A) protein was expressed highly in E. coli BL21 (DE3) by 0.5 mM IPTG induction and its yield could reach 650 mg/l in fermentation culture. The fusion protein in a form of inclusion body was then solubilized, refolded, and purified to a purity of 98% by cation exchange chromatography and size-exclusion chromatography. Four hundred and eighty milligrams protein of interest was obtained in per liter fermentation culture. The protein of interest was identified by SDS-PAGE and Western blot analysis, and great bioactivity of target protein to two cancer cell lines was confirmed by morphological changes and evaluated by MTT. The findings suggested that recombinant protein TAT-survivin (T34A) has a bright future in cancer therapy targeting towards survivin, and the efficient procedure of expression and purification may be useful for the mass production of this therapeutically important protein. PMID- 16260149 TI - Expression and purification of human placenta lactogen in Escherichia coli. AB - There are many growth factors secreted by placenta including growth hormone, placenta lactogen (PL), prolactin, follicle stimulating hormone, luteinizing hormone, thyroid stimulating hormone, and chorionic gonadotropin. For a systematic study of how these growth factors work together to result in the various biological functions and future clinical applications, it is needed to produce enough quantities of each protein. In this paper, we report the cloning of human PL (hPL) and expression by Escherichia coli (E. coli). Four kinds of expression vectors containing the hPL gene were transformed into several kinds of suitable host strains and grown at 37 and/or 30 degrees C. Determination of the yield of recombinant hPL by SDS-PAGE reveals that among the various conditions, pQE30-PL in E. coli strain M15[pREP4] expressed the largest amount of recombinant hPL at 37 degrees C. However, the expressed recombinant hPL was accumulated in inclusion body forms. The inclusion bodies were solubilized in 8M urea and purified by a His6 tagged affinity column under denaturing condition and the final yield of hPL was determined to be 48 mg/L. Intra-chain disulfide bonds could be formed either by oxidation in the refolding buffer or by air oxidation in the presence of urea. The biological activity was examined by the fact that hPL could stimulate erythroid maturation by the formation of hemoglobin in K-562 cells in the presence of erythropoietin. Initial optimization studies resulted in the production of 282.4 mg/L of hPL. PMID- 16260150 TI - An optimized system for expression and purification of secreted bacterial proteins. AB - In this report, we describe an optimized system for the efficient overexpression, purification, and refolding of secreted bacterial proteins. Candidate secreted proteins were produced recombinantly in Escherichia coli as Tobacco Etch Virus protease-cleavable hexahistidine-c-myc eptiope fusion proteins. Without regard to their initial solubility, recombinant fusion proteins were extracted from whole cells with guanidium chloride, purified under denaturing conditions by immobilized metal affinity chromatography, and refolded by rapid dilution into a solution containing only Tris buffer and sodium chloride. Following concentration on the same resin under native conditions, each protein was eluted for further purification and/or characterization. Preliminary studies on a test set of 12 secreted proteins ranging in size from 13 to 130 kDa yielded between 10 and 50 mg of fusion protein per liter of induced culture at greater than 90% purity, as judged by Coomassie-stained SDS-PAGE. Of the nine proteins further purified, analytical gel filtration chromatography indicated that each was a monomer in solution and circular dichroism spectroscopy revealed that each had adopted a well-defined secondary structure. While there are many potential applications for this system, the results presented here suggest that it will be particularly useful for investigators employing structural approaches to understand protein function, as attested to by the crystal structures of three proteins purified using this methodology (B.V. Geisbrecht, B.Y. Hamaoka, B. Perman, A. Zemla, D.J. Leahy, J. Biol. Chem. 280 (2005) 17243-17250). PMID- 16260151 TI - Purification and characterization of a recombinant anti-angiogenic kringle fragment expressed in Escherichia coli: Purification and characterization of a tri-kringle fragment from human apolipoprotein (a) (kringle IV (9)-kringle IV (10)-kringle V). AB - A kringle fragment (type IV (9)-IV (10)-V) from human apolipoprotein (a) (called LK68) was expressed in an inclusion body in Escherichia coli. The LK68 in this inclusion body was rendered soluble with urea, and efficiently refolded via oxidation in the presence of re-dox couple. The refolded LK68 was then purified via two steps of ion exchange chromatography, concentrated via preparative reversed-phase chromatography, and freeze-dried, at a final yield of approximately 30%. The purified LK68 exhibited profound affinity for lysine and fibrinogen, which suggests the proper folding of the kringle fragment, and also indicates that the native characteristics of apolipoprotein (a) were preserved. The purified LK68 was determined to be highly homogeneous upon reversed-phase HPLC analysis and size-exclusion HPLC analysis, in the presence of 20% (v/v) acetonitrile. However, on size-exclusion HPLC analysis without acetonitrile, it was determined to be somewhat heterogeneous, and this was corroborated by native analyses, including native PAGE and IEF. PMID- 16260152 TI - Specific detection of RNA on ultra-thin sections. AB - In this paper, we describe a specific method for ultrastructural detection of RNA. Our method is based in the bromination "in situ" of uridine residues in the RNA, which allows the detection of brominated RNA with specific antibodies against bromo-deoxyuridine. With this method we can achieve high specificity and resolution, and it can be applied to Epon or acrylic resin embedded material. PMID- 16260153 TI - White spot syndrome virus (WSSV) interaction with crayfish haemocytes. AB - WSSV particles were detected in separated granular cells (GCs) and semigranular cells (SGCs) by in situ hybridisation from WSSV-infected crayfish and the prevalence of WSSV-infected GCs was 5%, whereas it was 22% in SGCs. This indicates that SGCs are more susceptible to WSSV and that this virus replicated more rapidly in SGCs than in GCs and as a result the number of SGCs gradually decreased from the blood circulation. The effect of haemocyte lysate supernatant (HLS), containing the degranulation factor (peroxinectin), phorbol 12-myristate 13-acetate (PMA), the Ca(2+) ionophore A23187 on GCs from WSSV-infected and sham injected crayfish was studied. The results showed that the percentage of degranulated GCs of WSSV-infected crayfish treated with HLS or PMA was significantly lower than that in the control, whereas no significant difference was observed when treated with the Ca(2+) ionophore. It was previously shown that peroxinectin and PMA have a degranulation effect via intracellular signalling involving protein kinase C (PKC), whereas the Ca(2+) ionophore uses an alternative pathway. HLS treatment of GCs and SGCs from WSSV-infected crayfish results in three different morphological types: non-spread, spread and degranulated cells. The non-spread cell group from both GCs and SGCs after treatment with HLS had more WSSV positive cells than degranulated cells, when detected by in situ hybridisation. Taken together, it is reasonable to speculate that the PKC pathway might be affected during WSSV infection. Another interesting phenomenon was that GCs from non-infected crayfish exhibited melanisation, when incubated in L-15 medium, while no melanisation was found in GCs of WSSV-infected crayfish. However, the phenoloxidase activities of both sham- and WSSV-injected crayfish in HLS were the same as well as proPO expression as detected by RT-PCR. This suggests that the WSSV inhibits the proPO system upstream of phenoloxidase or simply consumes the native substrate for the enzyme so that no activity is shown. The percentage of apoptotic haemocytes in WSSV-infected crayfish was very low, but it was significantly higher than that in the sham-injected crayfish on day 3 or 5 post-infection. The TEM observation in haematopoietic cells (hpt cells) suggests that WSSV infect specific cell types in haematopoietic tissue and non-granular hpt cells seem more favourable to WSSV infection. PMID- 16260154 TI - Levels of consciousness and self-awareness: A comparison and integration of various neurocognitive views. AB - Quite a few recent models are rapidly introducing new concepts describing different levels of consciousness. This situation is getting confusing because some theorists formulate their models without making reference to existing views, redundantly adding complexity to an already difficult problem. In this paper, I present and compare nine neurocognitive models to highlight points of convergence and divergence. Two aspects of consciousness seem especially important: perception of self in time and complexity of self-representations. To this I add frequency of self-focus, amount of self-related information, and accuracy of self knowledge. Overall, I conclude that many novel concepts (e.g., reflective, primary, core, extended, recursive, and minimal consciousness) are useful in helping us distinguish between delicate variations in consciousness and in clarifying theoretical issues that have been intensely debated in the scientific literature--e.g., consciousness in relation to mirror self-recognition and language. PMID- 16260155 TI - fMRI resting state networks define distinct modes of long-distance interactions in the human brain. AB - Functional magnetic resonance imaging (fMRI) studies of the human brain have suggested that low-frequency fluctuations in resting fMRI data collected using blood oxygen level dependent (BOLD) contrast correspond to functionally relevant resting state networks (RSNs). Whether the fluctuations of resting fMRI signal in RSNs are a direct consequence of neocortical neuronal activity or are low frequency artifacts due to other physiological processes (e.g., autonomically driven fluctuations in cerebral blood flow) is uncertain. In order to investigate further these fluctuations, we have characterized their spatial and temporal properties using probabilistic independent component analysis (PICA), a robust approach to RSN identification. Here, we provide evidence that: i. RSNs are not caused by signal artifacts due to low sampling rate (aliasing); ii. they are localized primarily to the cerebral cortex; iii. similar RSNs also can be identified in perfusion fMRI data; and iv. at least 5 distinct RSN patterns are reproducible across different subjects. The RSNs appear to reflect "default" interactions related to functional networks related to those recruited by specific types of cognitive processes. RSNs are a major source of non-modeled signal in BOLD fMRI data, so a full understanding of their dynamics will improve the interpretation of functional brain imaging studies more generally. Because RSNs reflect interactions in cognitively relevant functional networks, they offer a new approach to the characterization of state changes with pathology and the effects of drugs. PMID- 16260156 TI - Males and females differ in brain activation during cognitive tasks. AB - To examine the effect of gender on regional brain activity, we utilized functional magnetic resonance imaging (fMRI) during a motor task and three cognitive tasks; a word generation task, a spatial attention task, and a working memory task in healthy male (n = 23) and female (n = 10) volunteers. Functional data were examined for group differences both in the number of pixels activated, and the blood-oxygen-level-dependent (BOLD) magnitude during each task. Males had a significantly greater mean activation than females in the working memory task with a greater number of pixels being activated in the right superior parietal gyrus and right inferior occipital gyrus, and a greater BOLD magnitude occurring in the left inferior parietal lobe. However, despite these fMRI changes, there were no significant differences between males and females on cognitive performance of the task. In contrast, in the spatial attention task, men performed better at this task than women, but there were no significant functional differences between the two groups. In the word generation task, there were no external measures of performance, but in the functional measurements, males had a significantly greater mean activation than females, where males had a significantly greater BOLD signal magnitude in the left and right dorsolateral prefrontal cortex, the right inferior parietal lobe, and the cingulate. In neither of the motor tasks (right or left hand) did males and females perform differently. Our fMRI findings during the motor tasks were a greater mean BOLD signal magnitude in males in the right hand motor task, compared to females where males had an increased BOLD signal magnitude in the right inferior parietal gyrus and in the left inferior frontal gyrus. In conclusion, these results demonstrate differential patterns of activation in males and females during a variety of cognitive tasks, even though performance in these tasks may not vary, and also that variability in performance may not be reflected in differences in brain activation. These results suggest that in functional imaging studies in clinical populations it may be sensible to examine each sex independently until this effect is more fully understood. PMID- 16260157 TI - Modulation of effective connectivity by cognitive demand in phonological verbal fluency. AB - Verbal fluency is a classic neuropsychological measure of language production. Phonological verbal fluency involves the generation of words beginning with a specified letter, and its functional neuroanatomy is comprised of a distributed network of regions which is modulated by cognitive load. In order to investigate the functional relationship of these regions, the effective connectivity was analyzed with covariance structural equation modeling under conditions of varying cognitive load. Significant path coefficients were evident between the anterior cingulate, left middle frontal gyrus, and precuneus. The left middle frontal gyrus showed a facilitory projection to the precuneus which had a suppressive influence on anterior cingulate activation. With increasing cognitive demand, the left middle frontal projection to the precuneus became suppressive, and the path coefficient from the precuneus to the anterior cingulate showed a marked diminution in strength. The path analysis suggests that the lead-in process for letter verbal fluency may primarily involve an orthographic visual strategy. The marked changes in path coefficients with the increased cognitive load may reflect the greater demands placed on executive function. The significant changes in path coefficient values with increased cognitive demand indicate the importance of accounting for task difficulty not only in the interpretation of brain activation maps but also for effective connectivity measurements. PMID- 16260158 TI - High frequency transformation of Cryptococcus neoformans and Cryptococcus gattii by Agrobacterium tumefaciens. AB - Cryptococcus neoformans and Cryptococcus gattii are the caus-ative agents of cryptococcal meningoencephalitis and are amenable to genetic manipulations, making them important models of pathogenic fungi. To improve the efficiency of Agrobacterium tumefaciens mediated transformation (ATMT) in C. neoformans, we optimized various co-cultivation conditions including incubation time and temperature, and bacteria to yeast ratio. ATMT was also applied to both serotypes (B and C) of C. gattii. Transformation efficiency by ATMT in C. neoformans was comparable to either electroporation or biolistic transformation and gave superior efficiencies in serotypes B and C, but unlike Saccharomyces cerevisiae, adenine auxotrophy did not increase ATMT efficiency in C. neoformans or C. gattii. All transformants tested were stable, with a majority containing only a single T-DNA insertion; however, homologous recombination was not observed. Additionally, we isolated adenine auxotrophs containing a single T-DNA insertion in the ADE2 gene for representative serotype B and C strains. PMID- 16260159 TI - Separation of 47Ti and 49Ti solid-state NMR lineshapes by static QCPMG experiments at multiple fields. AB - Experimental procedures are proposed and demonstrated that separate the spectroscopic contribution from both (47)Ti and (49)Ti in solid-state nuclear magnetic resonance spectra. These take advantage of the different nuclear spin quantum numbers of these isotopes that lead to different "effective" radiofrequency fields for the central transition nutation frequencies when these nuclei occur in sites with a significant electric field gradient. Numerical simulations and solid-state NMR experiments were performed on the TiO(2) polymorphs anatase and rutile. For anatase, the separation of the two isotopes at high field (21.1T) facilitated accurate determination of the electric field gradient (EFG) and chemical shift anisotropy (CSA) tensors. This was accomplished by taking advantage of the quadrupolar interaction between the EFG at the titanium site and the different magnitudes of the nuclear quadrupole moments (Q) of the two isotopes. Rutile, having a larger quadrupolar coupling constant (C(Q)), was examined by (49)Ti-selective experiments at different magnetic fields to obtain spectra with different scalings of the two anisotropic tensors. A small chemical shielding anisotropy (CSA) of -30 ppm was determined. PMID- 16260160 TI - Structure-based drug design to the discovery of new 2-aminothiazole CDK2 inhibitors. AB - N-(5-Bromo-1,3-thiazol-2-yl)butanamide (compound 1) was found active (IC50=808 nM) in a high throughput screening (HTS) for CDK2 inhibitors. By exploiting crystal structures of several complexes between CDK2 and inhibitors and applying structure-based drug design (SBDD), we rapidly discovered a very potent and selective CDK2 inhibitor 4-[(5-isopropyl-1,3-thiazol-2-yl)amino] benzenesulfonamide (compound 4, IC50=20 nM). The syntheses, structure-based analog design, kinases inhibition data and X-ray crystallographic structures of CDK2/inhibitor complexes are reported. PMID- 16260161 TI - Classification and comparison of ligand-binding sites derived from grid-mapped knowledge-based potentials. AB - We describe the application of knowledge-based potentials implemented in the MOE program to compare the ligand-binding sites of several proteins. The binding probabilities for a polar and a hydrophobic probe are calculated on a grid to allow easy comparison of binding sites of superimposed related proteins. The method is fast and simple enough to simultaneously use structural information of multiple proteins of a target family. The method can be used to rapidly cluster proteins into subfamilies according to the similarity of hydrophobic and polar fields of their ligand-binding sites. Regions of the binding site which are common within a protein family can be identified and analysed for the design of family-targeted libraries or those which differ for improvement of ligand selectivity. The field-based hierarchical clustering is demonstrated for three protein families: the ligand-binding domains of nuclear receptors, the ATP binding sites of protein kinases and the substrate binding sites of proteases. More detailed comparisons are presented for serine proteases of the chymotrypsin family, for the peroxisome proliferator-activated receptor subfamily of nuclear receptors and for progesterone and androgen receptor. The results are in good accordance with structure-based analysis and highlight important differences of the binding sites, which have been also described in the literature. PMID- 16260162 TI - The use of bronchodilators in the treatment of airway obstruction in elderly patients. AB - Ageing is associated with important anatomical, physiological and psychosocial changes that may have an impact on the management of obstructive airway diseases (asthma and chronic obstructive pulmonary disease (COPD)) and on their optimal therapy. Ageing-related modifications might be responsible for a different effectiveness of bronchodilators in the elderly patients as compared to younger subjects. Furthermore, the physiological involution of organs and the frequent comorbidity, often interfere with pharmacokinetics of bronchodilator drugs used in asthma and COPD. This review will focus on the use of bronchodilators in the elderly, with particular attention to the achievable goals and to rationale, utility and pitfalls in using the inhalation therapy in this age group. beta(2) agonists, anticholinergics and methylxanthines will be discussed and their side effects in the elderly will be considered. PMID- 16260163 TI - Two-tiered universal newborn screening strategy for severe combined immunodeficiency. AB - Outcomes for infants with severe combined immunodeficiency (SCID) would be improved by universal newborn screening, but there are not yet screening tests of sufficient accuracy for the disorder. In a pilot study, we assessed the ability of a two-tiered strategy to improve accuracy. Dried blood samples from patients were assessed with two tests for lymphopenia: interleukin-7, a T-cell growth cytokine, and TRECs, a byproduct of T-cell receptor recombination. IL-7 screening has a specificity of 96.1% and TRECs have a specificity of 92.3%. Combining these tests in a two-tiered strategy increases specificity to 100% (97-100% CI). Sensitivity was 85% for IL-7 screening and 100% for TREC screening. A two-tiered strategy may be of sufficient accuracy to enable universal SCID screening, and should be assessed in a prospective trial. PMID- 16260164 TI - Nutritional therapy improves growth and protein status of children with a urea cycle enzyme defect. AB - BACKGROUND: Poor growth has been described in patients with urea cycle enzyme defects treated with protein-restricted diets, while protein status is seldom reported. OBJECTIVE: To assess the effects of nutritional therapy with a medical food on growth and protein status of patients with a urea cycle enzyme defect. METHODS: A 6-mo multicenter outpatient study was conducted with infants and toddlers managed by nutrition therapy with Cyclinex-1 Amino Acid-Modified Medical Food with Iron (Ross Products Division, Abbott Laboratories, Columbus, OH). Main outcome variables were anthropometrics and plasma amino acids (selected), albumin, and transthyretin concentrations. RESULTS: Seventeen patients completed the study. Mean (+/-SE) baseline age was 11.30+/-3.20 months (median 4.40 months; range 0.22-38.84 months). Length and weight z-scores increased significantly during the 6-month study. Head circumference increased, but not significantly. Three patients were stunted and two were wasted (-2.0 z-score) at baseline while at study end, only one patient was both stunted and wasted. The majority of patients increased in length, head circumference, and weight z-scores during study. Mean (+/-SE) plasma albumin concentration increased from 34+/-2g/L at baseline to 38+/-1g/L at study end. Plasma transthyretin increased from a mean (+/-SE) of 177+/-13 mg/L at baseline to 231+/-15 mg/L at study end. No correlation was found between plasma NH(3) concentrations and medical food intake. Plasma NH(3) concentration was positively correlated with the percentage of Food and Agriculture Organization/World Health Organization/United Nations recommended protein ingested. CONCLUSIONS: Intakes of adequate protein and energy for age result in anabolism and linear growth without increasing plasma NH(3) concentrations. Medical food intakes did not correlate with plasma NH(3) concentrations. PMID- 16260165 TI - Pathways of galactose metabolism by galactosemics: evidence for galactose conversion to hepatic UDPglucose. AB - To determine if classic galactosemics have residual galactose-1-phosphate uridyltransferase (GALT) activity to explain their considerable ability to oxidize galactose over 24 h, we devised a method for assessing their ability to form hepatic UDPglucose (UDPglu), an intermediate in the normal Leloir pathway of galactose metabolism. The protocol involved the single oral administration of 7 mg/kg [2-13C]galactose concomitant with multiple small doses of acetaminophen with measurement of the extent of labeling of urinary acetaminophen glucuronide, the glucuronide moiety being formed from hepatic UDPglu. We performed the study lasting 24 h in two normal subjects and three classic galactosemics, two homozygous for the Q188R mutation and one compound for the Q188R/K258N mutation. The labeling and total excretion of acetaminophen glucuronide was measured in urine by nuclear magnetic resonance techniques. Concomitant with determination of label in the glucuronide measurement was made of galactose oxidation to 13CO2 and the 13C enrichment of plasma glucose. All of the galactosemic patients formed 13C enriched acetaminophen glucuronide indicating that they had converted the labeled galactose to [13C]UDPglu and that residual GALT or another pathway that forms UDPglu is present in hepatic tissue. Compared to the normal whose glucuronide labeling was rapid and short-lived that of the galactosemics was delayed and extended for a long period over 10 h. The extent of isotopic enrichment of glucuronide by galactosemics was comparable to the normals, resulting in a much greater conversion of galactose to UDPglu by the galactosemics. The labeling of the UDPglu pool was reflected by the rate of 13CO2 formation being rapid in the normal with peak labeling at 2-3 h with total oxidation of over 70% in 24 h. The oxidation of the galactosemics was slow with a broad peak of 13CO2 at 10 h and a total excretion of 25-39% of the [13C]galactose administered. The normal subjects formed highly enriched plasma glucose within 30 min while no enrichment of plasma glucose was detected until after 300 min in galactosemics. The exact pathway(s) of galactose metabolism by galactosemics to UDPglu remain to be determined. Their delineation may contribute to new approaches to therapeutic strategies for this enigmatic disorder. PMID- 16260167 TI - Symptomatic nosocomial urinary tract infection in very elderly patients. PMID- 16260166 TI - Chickenpox complications in Saudi Arabia: Is it time for routine varicella vaccination? AB - INTRODUCTION: Varicella zoster (chickenpox) infection is a common and benign disease of childhood. The predominance of uncomplicated cases in children tends to overshadow the morbidity associated with severe cases and the resultant hospitalization. OBJECTIVE: The objective was to establish the complication rate of chickenpox in the Saudi National Guard population over a 2(1/2)-year period. METHODS: All reported cases of chickenpox at the King Abdulaziz Medical City, Riyadh were prospectively followed for development of complications during the period between 1 June 2001 and 30 December 2003. RESULTS: A total of 3802 cases of chickenpox were reported and followed during the acute illness phase. The majority of cases, 2984 (78%), occurred in children less than 15 years of age. Among all chickenpox cases, 78 (2%) required hospitalization, and 50 patients developed 58 (1.5%) complications. Skin and soft tissue infections were the most common complications, 20 (34%), followed by pneumonia in 16 (28%), bacteremia in six (10%), encephalitis in four (7%) and necrotizing fasciitis in four (7%). Complications were less common in children (36/2984, 1.2%) as compared to adults (14/818, 1.7%) (p = 0.26). However, pneumonia (p = 0.03) and hospitalization (p = 0.004) were more common in adults as compared to children. The overall fatality rate was 0.05%; two patients died, one aged two months the other 28 years. CONCLUSION: These data provide a baseline for morbidity and mortality from chickenpox, and further support the inclusion of varicella vaccine in the routine childhood immunization program in Saudi Arabia. PMID- 16260168 TI - Coccidioidal pericarditis. PMID- 16260169 TI - Helicobacter pylori and Schistosoma japonicum co-infection in a Chinese population: helminth infection alters humoral responses to H. pylori and serum pepsinogen I/II ratio. AB - The effects of helminth infection on humoral IgG responses and clinical outcome of gastric Helicobacter pylori infection are unknown. IgG and IgG subclass responses to H. pylori and serum pepsinogen I/II ratio, a marker of gastric atrophy, were investigated in a Schistosoma japonicum prevalent Chinese population. H. pylori, CagA and IgG subclass responses were assayed by ELISA. Serum pepsinogen I and pepsinogen II were assayed by ELISA and the pepsinogen I/II ratio determined. In 150 subjects, infection with S. japonicum and H. pylori was 55.3% and 51.3%, respectively. H. pylori IgG titres and CagA seropositivity were significantly lower (P<0.05) in co-infected subjects, and differences in H. pylori IgG isotype responses were evident. In H. pylori positives, a significantly higher (P<0.05) pepsinogen I/II ratio was observed in co-infected subjects. The difference between S. japonicum positive and negative subjects was only evident in H. pylori CagA seronegative subjects. In conclusion, S. japonicum co-infection with H. pylori is associated with alterations in IgG responses to H. pylori and less gastric atrophy. PMID- 16260170 TI - Electroactive polymeric sensors in hand prostheses: bending response of an ionic polymer metal composite. AB - In stark contrast to the inspiring functionality of the natural hand, limitations of current upper limb prostheses stemming from marginal feedback control, challenges of mechanical design, and lack of sensory capacity, are well established. This paper provides a critical review of current sensory systems and the potential of a selection of electroactive polymers for sensory applications in hand prostheses. Candidate electroactive polymers are reviewed in terms of their relevant advantages and disadvantages, together with their current implementation in related applications. Empirical analysis of one of the most novel electroactive polymers, ionic polymer metal composites (IPMC), was conducted to demonstrate its potential for prosthetic applications. With linear responses within the operating range typical of hand prostheses, bending angles, and bending rates were accurately measured with 4.4+/-2.5 and 4.8+/-3.5% error, respectively, using the IPMC sensors. With these comparable error rates to traditional resistive bend sensors and a wide range of sensitivities and responses, electroactive polymers offer a promising alternative to more traditional sensory approaches. Their potential role in prosthetics is further heightened by their flexible and formable structure, and their ability to act as both sensors and actuators. PMID- 16260172 TI - An unusual pessary of dough and cocaine. PMID- 16260173 TI - Adding amino acids to the genetic repertoire. AB - Considerable progress has been made in expanding the number and nature of genetically encoded amino acids in Escherichia coli, yeast and mammalian cells in the past four years. To date, over 30 unnatural amino acids have been cotranslationally incorporated into proteins with high fidelity and efficiency by means of a unique codon and corresponding orthogonal tRNA-aminoacyl-tRNA synthetase pair. The incorporated amino acids contain spectroscopic probes, post translational modifications, metal chelators, photoaffinity labels and unique functional groups. The ability to genetically encode additional amino acids, beyond the common 20, provides a powerful approach for probing protein structure and function both in vitro and in vivo, as well as generating proteins with new or enhanced properties. PMID- 16260174 TI - Dendrimers as artificial enzymes. AB - Dendrimers are regular tree-like macromolecules accessible by chemical synthesis from a variety of building blocks. Their topology enforces a globular shape that offers a unique opportunity to design artificial enzymes. Catalytic groups such as metal complexes and cofactors can be placed at the dendrimer core to exploit microenvironment and selectivity effects of the dendritic shell. In a second approach, attaching catalytic groups in multiple copies at the end of the dendritic branches may lead to cooperativity effects. Finally, exploration of dendritic structural space by screening combinatorial libraries of peptide dendrimers for catalytic activity can lead to discovery of functional dendrimers with enzyme-like properties, in a process mimicking natural selection. PMID- 16260176 TI - Detection of initiating as well as promoting activity of chemicals by a novel cell transformation assay using v-Ha-ras-transfected BALB/c 3T3 cells (Bhas 42 cells). AB - Cell transformation assay using BALB/c 3T3 cells, C3H10T1/2 cells and others, can simulate the two-stage carcinogenesis utilized for formation of transformed foci. A sensitive cell transformation assay for tumor initiators as well as promoters has been developed using a v-Ha-ras-transfected BALB/c 3T3 cell line, Bhas 42; these cells are regarded as initiated in the two-stage paradigm of carcinogenesis. To distinguish between initiation and promotion, the initiation assay involves a 2-day treatment of low-density cells, obtained one day after plating, with a test chemical, and the promotion assay involves treatment of near confluent cells with a test chemical for a period of 12 days (Day 3-14). When Bhas 42 cells were treated with tumor initiators, N-methyl-N'-nitro-N nitrosoguanidine and 3-methylcholanthrene, transformed foci were induced in the initiation assay but not in the promotion assay. In contrast, tumor promoters, 12 O-tetradecanoylphorbol-13-acetate, lithocholic acid and okadaic acid, gave negative responses in the initiation assay but positive responses in the promotion assay. The results were reproducible with various treatment protocols. Sixteen polycyclic aromatic hydrocarbons were examined using both assays. Benzo[a]pyrene and 7,12-dimethylbenz[a]anthracene induced focus formation only in the initiation assay. Increase of focus formation was observed in the promotion assay with benzo[e]pyrene, benzo[ghi]perylene, 1-nitropyrene and pyrene. Benz[a]anthracene, benz[b]anthracene, chrysene and perylene showed positive responses in both initiation and promotion assays. Results of initiation and promotion assays of acenaphthylene, anthracene, coronene, 9,10 diphenylanthracene, naphthalene and phenanthrene were negative or equivocal. The present Bhas assays for the detection of either/both initiating and promoting activities of chemicals are sensitive and of high performance compared with other cell transformation assays. PMID- 16260175 TI - DNA repair capacity measured by high throughput alkaline comet assays in EBV transformed cell lines and peripheral blood cells from cancer patients and healthy volunteers. AB - We collected peripheral blood (PB) from 556 patients with various types of cancer who had undergone radiotherapy and from 81 healthy volunteers. We exposed whole PB and Epstein-Barr virus-transformed lymphoblastoid cell lines (EBLs) derived from the PB mononucleocytes to X-irradiation (5 Gy). Using the alkaline comet assay, we measured the immediate DNA damage and, at 15 min, the % residual damage. In PB, the immediate damage was similar in patients and healthy volunteers while the % residual damage (mean+/-S.D.) was significantly higher in patients with breast (54.3+/-A23.9), cervical (54.7+/-A23.9), head/neck (56.8+/ A24.4), lung (60.1+/-23.5), or esophageal cancers (59.5+/-A33.7) than in healthy donors (42.9+/-19.6) (P<0.05). We did not observe such differences in the EBV transformed cell lines. Thus, radiation sensitivity of fresh PB cells measured by the alkaline comet assay was related to cancer status. PMID- 16260177 TI - High prevalence of alpha 1 antitrypsin phenotypes in viral hepatitis B infected patients in Iran. AB - OBJECTIVE: Hepatitis B virus (HBV) infection is a major global public health problem. Approximately 2 billion people are infected worldwide and more than 350 million of these individuals are chronic carriers of HBV. Approximately 15-40% of infected patients will develop cirrhosis, liver failure, or hepatocellular carcinoma (HCC). Alpha 1 antitrypsin (AAT) deficiency is one of many factors that may be involved in abnormalities such as liver and lung disease, inflammatory joint diseases, and inflammatory eye diseases. In the present study, the role played by AAT in HBV infected individuals is analyzed. METHODS: AAT phenotyping and trypsin inhibitory capacity (TIC) experiments were performed on 281 HBV infected patients who were referred to Tehran and Zahedan Hepatitis Center for a period of 3 years from June 2001 to September 2003. The same tests were performed on 257 individuals who did not suffer from any systemic diseases (control group). The case group was subdivided into three groups: carrier (36.7%), chronic (50.5%), and cirrhotic (12.8%). RESULTS: The results showed that AAT phenotypes, MS, MZ, M(1)Z, and M(1)S, were significantly higher in the HBV group (p<0.01). In addition, there was a significant difference in AAT phenotypes (MS, MZ, and M(1)Z) among inactive carriers and individuals in the chronic and cirrhotic group (p<0.001). CONCLUSIONS: There is a high prevalence of moderate AAT (MS, M(1)S, and MV) and severely deficient (MZ and M(1)Z) phenotypes in Iranian HBV individuals. In addition, AAT deficiency might be a risk factor for infected HBV individuals progressing from the carrier stage to chronic and cirrhotic stages. PMID- 16260178 TI - Cost-effectiveness of radiofrequency ablation and surgical therapy for small hepatocellular carcinoma of 3cm or less in diameter. AB - BACKGROUND: Cost-effectiveness of radiofrequency ablation (RFA) was assessed in treatment of hepatocellular carcinoma (HCC). PATIENTS AND METHODS: During 5 years, 153 patients with HCC of 3cm or less received RFA, and 60 underwent surgery. Judgment after RFA therapy was classified into three grades: residual tumor (grade 1), necrotic area with a less safety margin of 5mm (grade 2), and necrosis with a safety margin of 5mm in all directions (grade 3). RESULTS: Local recurrence rates after RFA and surgery were 7.9% and 0% at the third year. The rates in patients with grades 2 and 3 after RFA were 18.7% and 1.2% at the third year, respectively (P=0.0005). Among 91 patients with grades 1 and 2 necrosis after initial therapy, 52 received additional ablation. Although local recurrence rate was 24.9% in 39 patients without additional therapy, the rates after therapy repetition were 10.9% in 21 patients with eventual grade 2 necrosis, and 0% in 31 patients with grade 3 (P=0.038). Median costs of single RFA, repeated RFA, and surgery were yen849,900, yen1,086,000, and yen1,745,100, respectively. Additional ablation reduced local recurrence by 20.7% at the cost of yen236,100. CONCLUSION: Cost-effectiveness of RFA in the treatment of small HCC was superior to that of surgery. PMID- 16260179 TI - Tiagabine enhances slow wave sleep and sleep maintenance in primary insomnia. AB - BACKGROUND AND PURPOSE: To evaluate the effect of tiagabine on sleep and next morning alertness and performance in adult patients with primary insomnia. PATIENTS AND METHODS: Patients with primary insomnia, as defined by Diagnostic and Statistical Manual of Mental Disorders--Fourth Edition (DSM-IV), received tiagabine 4, 8, 12, 16 mg, and placebo in a randomized, double-blind, five period, Latin square, crossover study. Efficacy was assessed using polysomnographic and self-report techniques; residual effects were evaluated using the Digit Symbol Substitution Test (DSST) and the Rey Auditory Verbal Learning Test (RAVLT). RESULTS: Fifty-eight patients (40f, 18m; mean age 46.6+/ 8.0 years) were randomized. Results showed a significant dose-dependent increase in slow wave sleep percentage with all tiagabine doses, a trend toward a dose dependent increase in total sleep time, and no effect on latency to persistent sleep. Wake after sleep onset also decreased in a dose-dependent manner, with the 16-mg dose differing significantly from placebo. The tolerability profiles of tiagabine 4 and 8 mg were similar to placebo. The most common adverse events reported following tiagabine 12 and 16 mg were dizziness and nausea. Residual effects were only apparent at 12- and 16-mg doses. CONCLUSIONS: Tiagabine increased slow wave sleep and reduced wake after sleep onset in a dose-dependent manner. Tiagabine dosages up to 8 mg did not compromise next-morning alertness and psychomotor performance in adult patients with primary insomnia. Further investigation of tiagabine doses up to 8 mg is warranted. PMID- 16260180 TI - A preliminary study of sleep-disordered breathing in major depressive disorder. AB - BACKGROUND AND PURPOSE: Individuals with obstructive sleep-disordered breathing (OSDB) commonly report symptoms of depression; however, the percentage of individuals with major depressive disorder (MDD) who experience OSDB is less clear. This study aimed to examine OSDB in a sample of individuals with MDD, unselected for sleep-related complaints, along a continuum of ventilatory and hypoxic abnormalities. PATIENTS AND METHODS: The overnight sleep-related breathing of 19 individuals with MDD and 15 non-depressed controls was recorded using an unattended nasal pressure-based home sleep monitoring device. The device recorded nasal airflow, breathing effort, heart rate, oxygen saturation, and body position. RESULTS: The two groups varied significantly on three sleep-related breathing variables: major flow-limitation events, major flow-limitation events accompanied by a desaturation, and average saturation throughout the evening; and these groups approached significance on minor flow-limitation events accompanied by a desaturation and average number of desaturations throughout the evening. Sleep-related breathing variables predicted accurate grouping in 81.3% of those with MDD and 80.6% of the non-depressed participants. CONCLUSIONS: These results suggest that OSDB may play a more important role in MDD than previously recognized. OSDB may contribute to or exacerbate the condition of someone predisposed to MDD, and the treatment of OSDB may ameliorate or possibly prevent depressive symptoms. PMID- 16260181 TI - Safety issues regarding new vaccines for tuberculosis, with an emphasis on post exposure vaccination. AB - There are several new types of vaccines that have become potential candidates for the prevention of tuberculosis. While these have been tested in the conventional animal models, less emphasis has been placed as yet on the development of safety testing procedures. This is particularly important if we are ever going to use such vaccines in a therapeutic or post-exposure mode, in which the risk of severe "Koch-like" reactions may be high. A further element that badly needs more research is the state of immunity already preexisting in the infected host and how vaccine induced immunity may interact with this. PMID- 16260182 TI - Structural biology of mycobacterial proteins: the Bangalore effort. AB - As part of an international effort and a national programme, structural analysis of mycobacterial proteins involved in recombination and repair, stringent response and protein synthesis has been undertaken, and work on proteins in a couple of metabolic pathways has been initiated. Already X-ray analysed are Mycobacterium tuberculosis and Mycobacterium smegmatis RecA and their nucleotide complexes, and different crystal forms of M. tuberculosis single-stranded DNA binding protein, M. smegmatis DNA binding protein from stationary phase cells and M. tuberculosis ribosome recycling factor. A comparative study involving these structures and those of similar proteins from other sources brings out the special features of the mycobacterial proteins, which are likely to be useful in selective inhibitor design. The structures provide insights into the plasticity of the molecules and its biological implications, and yield valuable information on their assembly and quaternary structure. They also provide leads for further structural investigations. PMID- 16260183 TI - Clinical link between MHC class II haplotype and interferon-beta (IFN-beta) immunogenicity. AB - Interferon-beta (IFN-beta) is currently the first-line therapy for the treatment of multiple sclerosis (MS). However, a significant percentage of MS patients develop anti-IFN-beta antibodies, which can reduce the efficacy of the drug. We describe an association between a common MHC class II allele (DRB1*0701), present in 23% of the patients studied, and the anti-IFN-beta antibody response. We identified IFN-beta epitopes using a peptide-binding assay with B cell lines expressing this allele. Moreover, epitope-specific activation responses obtained with peripheral blood mononuclear cells (PBMCs) from IFN-beta treated patients with the DRB1*0701 allele indicated a role for T-cell activation in IFN-beta immunogenicity. These results suggest that HLA typing of MS patients may provide an accurate screen for subjects who are likely to develop anti-IFN-beta antibodies and should therefore be considered for alternative therapies. In addition, elucidation of the factors underlying the anti-IFN-beta antibody response should accelerate the engineering of less immunogenic IFN-beta therapeutics. PMID- 16260184 TI - The role of LMO2 in development and in T cell leukemia after chromosomal translocation or retroviral insertion. AB - Chromosomal translocations are primary events in the development of leukemias, representing at least one genetic feature of the putative cancer stem cell. Studies of genes influenced by chromosomal translocations have yielded a vast amount of information about how cancer is initiated and maintained. In particular, acute leukemias have demonstrated that chromosomal translocations often involve transcription regulators that function by interacting with proteins and by controlling cell fate in the aberrant setting of the developing cancer cell. As a quintessential chromosomal translocation gene product, LMO2 has many properties that typify this class of molecule. In addition to its involvement in chromosomal translocations, the LMO2 gene was inadvertently activated in an X SCID gene therapy trial by retroviral insertion. New molecular therapies targeted directly at the LMO2 protein could have major impact as adjuncts to existing therapies or as therapeutics in their own right. In this review, we outline the current knowledge about LMO2 and some possible routes to develop reagents that might be possible macromolecular drugs in the future. PMID- 16260185 TI - High levels of persistent expression of alpha1-antitrypsin mediated by the nonhuman primate serotype rh.10 adeno-associated virus despite preexisting immunity to common human adeno-associated viruses. AB - Alpha1-antitrypsin (alpha1AT) deficiency is a genetic disorder causing emphysema if serum alpha1AT levels are <570 microg/ml. We have shown that intrapleural administration of an AAV5alpha1AT vector yielded persistent therapeutic alpha1AT serum levels. Since anti-AAV2 and -AAV5 antibodies prevalent in humans may limit the use of these common serotypes in gene therapy, we screened 25 AAV vectors derived from humans and nonhuman primates for alpha1AT expression following intrapleural administration to mice. The rhesus AAVrh.10 serotype yielded the highest levels and was chosen for further study. Following intrapleural administration, 77% of total body transgene expression was in the chest wall, diaphragm, lung, and heart. Intrapleural administration of AAVrh.10alpha1AT provided long-term, therapeutic alpha1AT expression in mice, although higher doses were required to achieve therapeutic levels in female mice than in male mice. Intrapleural administration of AAVrh.10alpha1AT produced the same levels in AAV2/AAV5-preimmune and naive mice. In mice administered with AAV5alpha1AT and subsequently "boosted" with the AAVrh.10alpha1AT vector, serum levels were increased by 300%. These data indicate that AAVrh.10 is the most effective known AAV vector for intrapleural gene delivery and has the advantage of circumventing human immunity to AAV. PMID- 16260186 TI - KDE Bioscience: platform for bioinformatics analysis workflows. AB - Bioinformatics is a dynamic research area in which a large number of algorithms and programs have been developed rapidly and independently without much consideration so far of the need for standardization. The lack of such common standards combined with unfriendly interfaces make it difficult for biologists to learn how to use these tools and to translate the data formats from one to another. Consequently, the construction of an integrative bioinformatics platform to facilitate biologists' research is an urgent and challenging task. KDE Bioscience is a java-based software platform that collects a variety of bioinformatics tools and provides a workflow mechanism to integrate them. Nucleotide and protein sequences from local flat files, web sites, and relational databases can be entered, annotated, and aligned. Several home-made or 3rd-party viewers are built-in to provide visualization of annotations or alignments. KDE Bioscience can also be deployed in client-server mode where simultaneous execution of the same workflow is supported for multiple users. Moreover, workflows can be published as web pages that can be executed from a web browser. The power of KDE Bioscience comes from the integrated algorithms and data sources. With its generic workflow mechanism other novel calculations and simulations can be integrated to augment the current sequence analysis functions. Because of this flexible and extensible architecture, KDE Bioscience makes an ideal integrated informatics environment for future bioinformatics or systems biology research. PMID- 16260187 TI - JAK-STAT pathway in cardiac ischemic stress. AB - In our quest for comprehensive protection of ischemic myocardium, both basic and clinical studies have lead us to examine signal transduction pathways involved in ischemia-reperfusion injury for potential therapeutic targets. In this review, we have highlighted the importance of the JAK-STAT pathway in modulating ischemia reperfusion injury. The mechanisms linking glucose metabolism, angiotensin II, with JAK-STAT pathway in ischemic injury are explored in this review. Clearly, the studies discussed in this review provide rationale for the design and synthesis of selective blockers of JAK-STAT pathway as potential therapeutic adjuncts in protecting ischemic myocardium. PMID- 16260188 TI - Optimal number of repeated measures and group sizes in clinical trials with linearly divergent treatment effects. AB - The effect of number of repeated measures on the variance of the generalized least squares (GLS) treatment effect estimator is considered assuming a linearly divergent treatment effect, equidistant time-points and either a fixed number of subjects or a fixed study budget. The optimal combination of group sizes and number of repeated measures is calculated by minimizing this variance subject to a linear cost function. For a fixed number of subjects, the variance of the GLS treatment effect estimator can be decreased by adding intermediate measures per subject. This decrease is relatively large if a) the covariance structure is compound symmetric or b) the structure approaches compound symmetry and the correlation between two repeated measures does not exceed 0.80, or c) the correlation between two repeated measures does not exceed 0.60 if the time-lag goes to zero. In case the sample sizes and number of repeated measures are limited by budget constraints and the covariance structure includes a first-order auto-regression part, two repeated measures per subject yield highly efficient treatment effect estimators. Otherwise, it is more efficient to have more than two repeated measures. If the covariance structure is unknown, the optimal design based on a first-order auto-regressive structure with measurement error is preferable in terms of robustness against misspecification of the covariance structure. The numerical results are illustrated by three examples. PMID- 16260189 TI - Gene expression changes reveal patterns of aging in the rat digestive tract. AB - Similarly to other organs, the human digestive system is adversely affected by aging presenting physiologic manifestations that include compromised absorption and secretion, decreased motility, weakened mucosal barrier and as well as a high incidence of colon cancer. As biomedical advances enable the population to live longer, our understanding of molecular events that govern aging and disease states is enhanced through methodical analyses of temporal tissue-specific gene expression profiles. Recently, DNA microarray analyses have been employed to examine age-associated transcriptional profiles in the mammalian digestive tract. Gene expression patterns revealed that the magnitude and trend of age-associated changes differ in the rat colon and duodenum. Interestingly, the expression of genes involved in energy-generating metabolic pathways was decreased in the duodenum and increased in the colon. Microarray analyses detected modulations in expression of genes associated with compromised intestinal function and propensity for colon cancer in the aged population. Furthermore, altered expression was observed for certain genes implicated in governance of aging and lifespan in other organisms suggesting intriguing commonalities across species. Thus, these studies demonstrated feasibility and usefulness of DNA microarrays for identifying pathways involved in the molecular pathophysiology of the aging process and lifespan control in complex organisms. PMID- 16260190 TI - Quantification of tipranavir in human plasma by high-performance liquid chromatography with UV detection. AB - A simple method for the quantification of tipranavir, a new non-peptidic protease inhibitor, was developed. An internal standard, prazepam, was added to 100 microl of plasma before a liquid-liquid extraction by 3 ml of tert-butyl methyl ether. The extracts were evaporated to dryness and reconstituted with 100 microl of mobile phase before being injected in the chromatographic system. The separation was made on a C8 column using sodium acetate buffer (pH 5):methanol:acetonitrile (35:30:35, v/v/v) as mobile phase. The detection was performed at a wavelength of 260 nm. The method was linear and has been validated over a concentration range of 2-80 mg/l. The mean precision and accuracy of the method were respectively, 10.5 and -9.1%. The mean recovery was 70.8%. PMID- 16260192 TI - Influence of storage temperature and freezing time on histamine level in the European anchovy Engraulis encrasicholus (L., 1758): A study by capillary electrophoresis. AB - Histamine content in fish may increase by decarboxylation of free histidine to values that can be toxic, if storage conditions are not well controlled. We have studied the influence of storage temperature and time of freezing on histamine formation in the anchovy, Engraulis encrasicholus (L., 1758), for which little information is available. Analysis, carried out by capillary zone electrophoresis (CZE) without sample pre-treatment, was very simple, fast and reproducible. Results indicate that temperatures above 20 degrees C notably increase histamine production, whereas freezing can clearly prevent or slow down the process. PMID- 16260191 TI - Determination of the heat shock protein 90 inhibitor 17-dimethylaminoethylamino 17-demethoxygeldanamycin in plasma by liquid chromatography-electrospray mass spectrometry. AB - A rapid method was developed for the quantitative determination of the novel heat shock protein 90 inhibitor, 17-dimethylaminoethylamino-17-demethoxygeldanamycin (17-DMAG; NSC707545), in human plasma. Calibration curves were constructed, and were analyzed using a weight factor proportional to the nominal concentration. Sample pretreatment involved a one-step extraction with ethyl acetate of 0.5-ml samples. The analysis was performed in the range of 1-100 ng/ml on a column (75 mm x 2.1 mm internal diameter with 3.5 microm C18 particle size), using 55% methanol in water containing formic acid as the mobile phase. The column effluent was monitored by mass spectrometry with positive electrospray ionization. The values for precision and accuracy were always <8% and <10% relative error, respectively. The method was successfully applied to examine the pharmacokinetics of 17-DMAG in a cancer patient. PMID- 16260193 TI - Alternative splicing: a new drug target of the post-genome era. AB - Alternative splicing allows for the creation of multiple distinct mRNA transcripts from a given gene in a multicellular organism. Pre-mRNA splicing is catalyzed by a multi-molecular complex, including serine/arginine-rich (SR) proteins, which are highly phosphorylated in living cells, and thought to play crucial roles in spliceosomal formation and in the regulation of alternative splicing. Recently, reports of low molecular compounds, which alter splicing pattern of genes, have been accumulated. A benzothiazole compound TG003, a kinase inhibitor that targets Clk1 and Clk4, suppressed dissociation of nuclear speckles, altered the splicing patterns, and rescued the embryonic defects induced by excessive Clk activity. The emerging inhibitors of the signal transduction pathways regulating pre-mRNA alternative splicing may open the way to therapies against diseases caused by missplicing. PMID- 16260194 TI - Characterization and crystallization of human DPY-30-like protein, an essential component of dosage compensation complex. AB - Human DPY-30-like is a homolog of C. elegans DPY-30. DPY-30 is an essential component of dosage compensation machinery and loss of dpy-30 activity results in XX-specific lethality. In XO animals, DPY-30 is required for developmental processes other than dosage compensation. In yeast, the homolog of DPY-30, Saf19p, functions as a member of histone 3 lysine 4 methylation complex, which is the key part of epigenetic developmental control. In this report, human DPY-30 like protein was overexpressed and purified with the goal of structure determination. It was crystallized at 291 K in hanging drops by the vapour diffusion technique from a precipitant solution consisting of (NH4)2SO4 (1.5-2.0 M), Tris-HCl (0.1 M, pH 8.0). The crystal diffracted to 2.7 A resolution at 100 K in-house and belongs to the space group P4(1)2(1)2 or P4(3)2(1)2 with unit-cell parameters of a=b=74.5 A, c=87.0 A, alpha=beta=gamma=90.0 degrees. The asymmetric unit contains two molecules with 49% solvent content. We also analyzed its biochemical and biophysical characterizations. Efforts are now under way to determine the molecular structure of the DPY-30-like. These studies will open a new avenue towards the structure-based functional analysis of human DPY-30-like and dosage compensation machinery. PMID- 16260195 TI - Age-related change in the damage morphology of human cortical bone and its role in bone fragility. AB - Application of cyclic loading results in the formation of distinct strain dependent microdamage morphologies. It is still unknown; however, how the morphology of microdamage affects age-related increase in bone fragility. In this study, four-point bending fatigue tests were conducted on aging human bone (age 26 to 89) in conjunction with histological evaluation of the resultant tensile (diffuse damage) and compressive (linear microcracks) damage to identify the damage morphologies associated with an increase in age-related bone fragility. The results demonstrate that young donors (38 +/- 9 years) had a longer fatigue life (P < 0.05) and formed more diffuse damage than the older donors (82 +/- 5 years) (P < 0.05). In contrast, old donors had a shorter fatigue life and formed more linear microcracks than the younger donors (P < 0.05). Linear microcracks were longer in older than in younger donors (P < 0.05) and were associated with weak lamellar interfaces. Areas of diffuse damage were, however, larger in younger than in older donors (P < 0.05), and these showed no relationship with the lamellar arrangement of bone. These findings show, for the first time, that the propensity of bone to form a particular damage morphology is subject to change with age and that the propensity of young donors to form diffuse damage over interlamellae linear microcracks plays a critical role in the ability of bone to dissipate energy and resist a catastrophic fracture. Age-related changes in damage morphology may therefore be an important contributor to the increased bone fragility in the elderly. PMID- 16260196 TI - Dexamethasone to improve maternal outcome in women with hemolysis, elevated liver enzymes, and low platelets syndrome. PMID- 16260197 TI - Dexamethasone treatment does not improve the outcome of women with HELLP syndrome: a double-blind, placebo-controlled, randomized clinical trial. AB - OBJECTIVE: The purpose of this study was to determine the efficacy of dexamethasone for treatment of HELLP (hemolysis, elevated liver enzymes and low platelet count) syndrome. STUDY DESIGN: A prospective, double-blind clinical trial was conducted among 132 women with HELLP syndrome who were assigned randomly to treatment or placebo groups. Pregnant women in the experimental group received 10-mg doses of dexamethasone intravenously every 12 hours until delivery and 3 additional doses after delivery. Puerperal women received 3 10-mg doses of dexamethasone after delivery. The same schedule was used in the placebo group. The main outcome variable was the duration of hospitalization. In addition, we evaluated treatment effects on the time to recovery of laboratory and clinical parameters and on frequency of complications. RESULTS: The mean duration of hospitalization of patients who received dexamethasone therapy was shorter than in the placebo group (6.5 vs 8.2 days), but this difference was not statistically significant (P = .37). No significant differences were found in the time to recovery of platelet counts (hazard ratio, 1.2; 95% CI, 0.8-1.8), lactate dehydrogenase (hazard ratio, 0.9; 95% CI, 0.5-1.5), aspartate aminotransferase (hazard ratio, 0.6; 95% CI, 0.4-1.1) and to the development of complications. The results were found in both pregnant and puerperal women. CONCLUSION: The results of this investigation do not support the use of dexamethasone for treatment of HELLP syndrome. PMID- 16260198 TI - WHI clinical trial revisit: imprecise scientific methodology disqualifies the study's outcomes. AB - We analyzed The Women's Health Initiative (WHI) Study because it had a significant impact on clinical practice, both nationally and internationally. However, despite the widespread public and professional awareness of the results, an independent, nonbiased analysis of the quality of the methodology of the study has not been available. We find the study design and its execution question the validity of the results, making it difficult to apply the WHI results to healthy postmenopausal women, different ethnic groups, or as general postmenopausal prevention. PMID- 16260200 TI - Evidence-based surgery for cesarean delivery. AB - OBJECTIVE: The purpose of this study was to provide evidenced-based guidance for surgical decisions during cesarean delivery. STUDY DESIGN: We performed MEDLINE, PubMed, EMBASE, and COCHRANE searches with the terms cesarean section, cesarean delivery, cesarean, pregnancy, randomized trials, and each technical aspect of cesarean delivery. All randomized trials that covered a surgical aspect of cesarean delivery were included in the review. Each surgical step of cesarean delivery was reviewed separately. RESULTS: US Preventive Services Task Force recommendations favor blunt uterine incision expansion, prophylactic antibiotics (either ampicillin or first-generation cephalosporin for just 1 dose), spontaneous placental removal, non-closure of both visceral and parietal peritoneum, and suture closure or drain of the subcutaneous tissue when thickness is > or =2 cm. CONCLUSION: Cesarean delivery techniques that are supported by good quality recommendations should be performed routinely. All technical aspects that have recommendations with lower quality should be researched with adequately powered and designed trials. PMID- 16260201 TI - Uterine artery embolization versus hysterectomy in the treatment of symptomatic uterine fibroids (EMMY trial): peri- and postprocedural results from a randomized controlled trial. AB - OBJECTIVE: This was a randomized controlled trial to evaluate the safety of uterine artery embolization (UAE) compared with hysterectomy. STUDY DESIGN: Twenty-eight Dutch hospitals recruited 177 patients with symptomatic uterine fibroids and menorrhagia who were eligible for hysterectomy. Patients were randomized to UAE (n = 88) or hysterectomy (n = 89). In this paper we evaluate the peri- and postprocedural complications, length of hospital stay, unscheduled visits, and readmission rates up to 6 weeks' post-intervention. Analysis was by intention to treat. RESULTS: Bilateral UAE failure occurred in 4 patients (4.9%). Major complications occurred in 4.9% (UAE) and 2.7% (hysterectomy) of cases (P = .68). The minor complication rate from discharge until 6 weeks after was significantly higher in the UAE group than in the hysterectomy group (58.0% vs 40.0%; RR 1.45 [1.04-2.02]; P = .024). UAE patients were more often readmitted (11.1% vs 0%; P = .003). Total length of hospital stay was significantly shorter in UAE patients (mean [SD]: 2.5 [2.7] vs 5.1 [1.3], P < .001). CONCLUSION: UAE is a procedure similar to hysterectomy with a low major complication rate and with a reduced length of hospital stay. Higher readmission rates after UAE stress the need for careful postprocedural follow-up. PMID- 16260202 TI - Ovarian cancer screening in the Prostate, Lung, Colorectal and Ovarian (PLCO) cancer screening trial: findings from the initial screen of a randomized trial. AB - OBJECTIVE: Ovarian cancer screening with transvaginal ultrasound (TVU) and CA-125 was evaluated in the Prostate, Lung, Colorectal and Ovarian (PLCO) Trial. STUDY DESIGN: This was a randomized controlled trial of screening versus usual care. Baseline screening results are reported. RESULTS: Of 39,115 women randomized to receive screening, 28,816 received at least 1 test. Abnormal TVU was found in 1338 (4.7%), and abnormal CA-125 in 402 (1.4%). Twenty-nine neoplasms were identified (26 ovarian, 2 fallopian, and 1 primary peritoneal neoplasm). Nine were tumors of low malignant potential and 20 were invasive. The positive predictive value for invasive cancer was 3.7% for an abnormal CA-125, 1.0% for an abnormal TVU, and 23.5% if both tests were abnormal. CONCLUSION: The effect of screening on ovarian cancer mortality in the PLCO cohort has yet to be evaluated and will require longer follow-up. Screening identified both early- and late stage neoplasms, and the predictive value of both tests was relatively low. PMID- 16260203 TI - Endometrial cancer in women 45 years of age or younger: a clinicopathological analysis. AB - OBJECTIVE: The purpose of this study was to evaluate the experience with endometrial carcinoma in women < or =45 years of age at Ochsner Clinic Foundation, New Orleans, La. STUDY DESIGN: We evaluated the clinical history, treatment, and follow-up of 38 women < or =45 years of age diagnosed with endometrial cancer. RESULTS: Thirty-eight patients received primary treatment for endometrial cancer: stage I, 32 (84.2%); stage II, 1 (2.6%); stage III, 4 (10.5%); stage IV, 1 (2.6%). Tumors were well differentiated in 20 (52.6%), moderately differentiated in 10 (26.3%), and poorly differentiated in 8 (21.1%). At end of study period 32 women (84.2%) were alive with no evidence of disease, 5 had died of recurrent disease, and 1 died of metastatic breast cancer. CONCLUSION: Patients < or =45 years of age had lower incidence of advanced stage disease, higher degree of tumor differentiation, and better prognosis compared to patients older than 45 years. PMID- 16260205 TI - Uterine innervation after hysterectomy for chronic pelvic pain with, and without, endometriosis. AB - OBJECTIVE: Chronic pelvic pain is associated with a wide range of clinical conditions that include endometriosis. The precise cause, mechanisms of pain, and natural history are imprecise. Patterns of uterine innervation have been studied after hysterectomy for chronic pelvic pain with and without endometriosis. STUDY DESIGN: Tissue blocks were taken from the lower one half of the uterus after hysterectomy for advanced endometriosis (n = 16 specimens; group 1) and for chronic pelvic pain without endometriosis (n = 15 specimens; group 2). The control group consisted of uteri that were removed for painless gynecologic conditions (n = 25 specimens; group 3). Tissue sections from the lower one half of the uterus were stained with anti-S100 to demonstrate patterns of innervation, and nerve fiber profiles were counted by standardized techniques; qualitative differences were also recorded. RESULTS: In uteri from women with advanced endometriosis, there were increased numbers of nerve fiber profiles compared with control specimens (group 1 vs group 3; P = .0013, Mann Whitney U test). There were also increased numbers of nerve fiber profiles in uteri that were associated with chronic pelvic pain without endometriosis (group 2 vs group 3; P = .04, Mann Whitney U test). There were no differences in nerve fiber count in uteri from groups 1 and 2 (P = .35, Mann Whitney U test). Comparing both groups of uteri with controls (groups 1 and 2 vs 3) demonstrated marked differences in nerve fiber counts (P = .002, Mann Whitney U test). Two distinctive patterns of reinnervation that were observed: disruption of nerve bundles (collateral sprouting with microneuroma formation) and ingrowth around blood vessels (perivascular nerve fiber proliferation). There were increased numbers of microneuromas (groups 1 and 2 vs 3; P = .001, chi-squared test with Yates correction) and perivascular nerve fiber proliferation (groups 1 and 2 vs 3; P = .008, chi-squared test with Yates correction) in the myometrium in chronic pelvic pain with, and without, endometriosis compared with the control group. CONCLUSION: Nerve fiber proliferation and other features of reinnervation have been observed in the isthmic regions of uteri that were removed at hysterectomy for chronic pelvic pain with and without endometriosis. There were no quantitative differences between the groups with chronic pelvic pain and endometriosis. These observations provide an alternative explanation for the source of pain and other clinical symptoms in these clinical settings. PMID- 16260204 TI - Laparoscopic staging in patients with incompletely staged cancers of the uterus, ovary, fallopian tube, and primary peritoneum: a Gynecologic Oncology Group (GOG) study. AB - OBJECTIVE: The purpose of this study was to determine the feasibility of laparoscopically staging patients with incompletely staged cancers of the uterus, ovary, fallopian tube, and primary peritoneum, and to evaluate related effects. STUDY DESIGN: Patients without evidence of metastatic disease had laparoscopic bilateral para-aortic and pelvic lymph node dissection. Other procedures were individualized based on extent of the primary surgery; laparotomy was undertaken for identified resectable disease. RESULTS: Ninety-five eligible patients were entered on 2 Gynecologic Oncology Group (GOG) protocols. Eleven were excluded. Fifty-eight patients (69%) underwent complete endoscopic staging with photographic documentation. Nine others (10%) were incompletely staged. Seventeen patients (20%) had laparotomy. In patients undergoing laparoscopy, 6% had bowel complications; 11% were found to have more advanced disease. Hospital stay was significantly shorter with laparoscopy alone (3 vs 6 days, P = .04). CONCLUSION: Interval laparoscopic staging of gynecologic malignancies can be successfully undertaken in selected patients, but laparotomy for adhesions or metastatic disease and risk of visceral injury may be anticipated. PMID- 16260206 TI - Maternal complications with vaginal birth after cesarean delivery: a multicenter study. AB - OBJECTIVE: This study was undertaken to determine incidence and risk factors for uterine rupture in women attempting vaginal birth after cesarean delivery (VBAC) in a wide range of hospital settings. STUDY DESIGN: We performed a case-control study nested within a cohort of women who have had a prior cesarean to determine the incidence and risk factors for uterine rupture in women attempting VBAC. RESULTS: The incidence rate of uterine rupture in those who attempt VBAC was 9.8 per 1000. A prior vaginal delivery was associated with a lower risk of uterine rupture (adjusted odds ratio [OR] = 0.40, 95% CI 0.20-0.81). Although prostaglandins alone were not associated with uterine rupture, sequential use of prostaglandin and pitocin was associated with uterine rupture (adjusted OR = 3.07, 95% CI 0.98-9.88). CONCLUSION: Women with a prior cesarean should be offered VBAC, and women with a prior cesarean and prior vaginal delivery should be encouraged to VBAC. Although other studies have suggested that prostaglandins should be avoided, we suggest that inductions requiring sequential agents be avoided. PMID- 16260207 TI - Increased intrauterine frequency of Ureaplasma urealyticum in women with preterm labor and preterm premature rupture of the membranes and subsequent cesarean delivery. AB - OBJECTIVE: The purpose of this study was to evaluate the prevalence of microbial invasion of the amniotic cavity at the time of preterm cesarean delivery for therapy-resistant preterm labor or preterm premature rupture of membranes, which are events that commonly are induced by infection, and to compare this group of patients with a group of patients who underwent preterm cesarean delivery for indications other than preterm labor or preterm premature rupture of membranes. STUDY DESIGN: We studied 207 consecutive women between 23 and 34 weeks of gestation who underwent cesarean delivery. These patients were divided into 3 groups according to the indication for cesarean delivery: patients with preterm labor (group 1), patients with preterm premature rupture of membranes (group 2), and patients with other indications (group 3). In the course of the surgical procedure, amniotic fluid, amniotic membrane, and placental tissue specimens were collected for the detection of pathogens. RESULTS: Ureaplasma urealyticum was detected in 43.9% (58/132) of the patients of groups 1 and 2, with no significant difference between these 2 subgroups. In group 3, which served as the comparison group, Ureaplasma urealyticum was isolated in only 2.7% (2/75) of the patients. Ureaplasma urealyticum as a single pathogen was more frequent than all obligate pathogens together (43.9% vs 39.3%). CONCLUSION: Our results provide evidence for an association between intrauterine colonization with Ureaplasma urealyticum and both therapy-resistant preterm labor and preterm premature rupture of membranes. PMID- 16260208 TI - Persistance of adverse obstetric and neonatal outcomes in monochorionic twins after exclusion of disorders unique to monochorionic placentation. AB - OBJECTIVE: This study was undertaken to assess obstetric and neonatal outcomes in dichorionic twins and monochorionic-diamniotic twins after exclusion of twin-to twin transfusion syndrome and twin reversed arterial perfusion sequence. STUDY DESIGN: Data from a tertiary center were collected in twin gestations between 1994 and 2002. Chorionicity was defined by standard echographic criteria and placental examination at delivery. Neonatal outcomes were compared between monochorionic and dichorionic gestations. RESULTS: This study included 503 women: 378 (75%) dichorionic and 125 (25%) monochorionic twin gestations. Monochorionic twin gestations had a higher risk of preterm deliveries between 30 and 34 weeks' gestation than pregnancies with dichorionic twins (P < .01). Monochorionic twins had a higher number of birth weight less than 10th percentile (P < .001) discordancy 25% or greater (P < .02), admission to neonatal intensive care unit (P < .03), and intraventricular hemorrhage grade 3 and 4 (P < .007) than dichorionic twins even after adjusting for gestational age. CONCLUSION: Monochorionic diamniotic twins have a higher risk of perinatal complications than dichorionic twin gestations, even after exclusion of disorders unique to monochorionic placentation. PMID- 16260209 TI - Hypertensive disease in pregnancies complicated by systemic lupus erythematosus. AB - OBJECTIVE: The purpose of this study was to assess the percentage of hypertensive disease in pregnancies complicated by systemic lupus erythematosus at a single institution. STUDY DESIGN: We conducted a retrospective analysis of medical records between 1992 and 2003 of 68 pregnancies that were complicated by systemic lupus erythematosus from 48 parturients. Patients were categorized into 3 groups: no chronic hypertension (n = 49 women), chronic hypertension-no medication (n = 6 women) and chronic hypertension-treated (n = 13 women). Analyses of variance (with Tukey-Kramer adjusted follow-up evaluation) and chi-squared/Fisher's exact tests were used for the analyses of continuous and categoric variables, respectively. Significance was defined by a probability value of < or = .05. RESULTS: Chronic hypertension complicated 28% of systemic lupus erythematosus pregnancies. Mean systolic blood pressures at intake were significantly different between the normotensive and no chronic hypertension groups and between the chronic hypertension-no medication and chronic hypertension-treated groups; the differences in diastolic pressures reached significance only between the no chronic hypertension and the chronic hypertension-treated groups. Maternal age, gestational age at delivery, birth weight, lowest platelet count, and highest serum creatinine levels were similar between the hypertensive and the nonhypertensive groups. There were no differences in the percentage of aspirin or heparin treatments among the groups, but the percentage of the chronic hypertension-treated group who received steroids was significantly greater than the percentage of women who received steroids in the other 2 groups (P < .05). Preeclampsia developed in 23% of the no chronic hypertension pregnancies and in 32% of the hypertensive pregnancies (P = .54). When pregnancies that were treated with prednisone (n = 34 pregnancies) were compared with those pregnancies that were managed with other agents (n = 34 pregnancies), the percentages of preeclampsia were similar (26% and 24%, respectively; P = .78). CONCLUSION: The percentage of parturients with systemic lupus erythematosus in whom preeclampsia develops is increased, regardless of the presence of underlying chronic hypertension. Prednisone therapy was not associated with a higher risk of preeclampsia in this series. PMID- 16260210 TI - Gestational age-specific predicted risk of neonatal respiratory distress syndrome using lamellar body count and surfactant-to-albumin ratio in amniotic fluid. AB - OBJECTIVE: This study was undertaken to study the statistical correlation between lecithin/sphingomylein (L/S) ratio, percent phosphatidylglycerol (%PG), lamellar body count (LBC), and surfactant-to-albumin ratio (TDx-FLM(II)) in amniotic fluid (AF); and derive gestational age-specific (GA) predicted risk of neonatal respiratory distress syndrome (RDS) for LBC and TDx-FLM(II). STUDY DESIGN: AF specimens (238) were collected by transabdominal amniocentesis. L/S ratio, %PG, LBC, and TDx-FLM(II) were determined by established procedures. RDS diagnosis was ascertained by a neonatalogist, and statistical analyses were performed with the use of the SPSS software program (SPSS Inc, Chicago, Ill). RESULTS: Significant correlation was obtained among the 4 variables (L/S ratio, %PG, LBC, and TDx FLM(II)). Independent linear regression analyses between L/S ratio versus LBC and TDx-FLM(II) provided acceptable correlation. Multiple regression analysis showed a significant (P < .001) contribution from TDx-FLM(II) and GA for predicting the L/S ratio. Receiver operating characteristic curve analysis provided the immature cutoffs (LBC = < 30.0 x 10(3)/microL; TDx-FLM(II) = < 40.0 mg/g). Total accuracy (either positive or negative) for RDS was similar for LBC (75.5%) and TDx-FLM(II) (76.7%). CONCLUSION: LBC and TDx-FLM(II) are equally accurate. GA-specific predicted risk of RDS by both tests significantly eliminated L/S ratio identified false positive cases of fetal lung maturity. PMID- 16260211 TI - The mean weekly increment of amniotic fluid TDx-FLM II ratio is constant during the latter part of pregnancy. AB - OBJECTIVE: The purpose of this study was to determine the mean weekly increment in amniotic fluid TDx-FLM II ratio during the latter part of pregnancy. STUDY DESIGN: All women who underwent > 1 amniocentesis for the determination of fetal lung maturity between 1998 and 2004 were identified retrospectively. Clinical information and TDx-FLM II ratios were collected from the participant's chart and analyzed. RESULTS: The gestational age of all participants at the first TDx-FLM II test was 31.2 to 37.5 weeks (mean, 34.7 +/- 1.4 weeks of gestation). The median interval between the 2 tests was 7 days (range, 5-36 days). We found that the mean weekly increment of TDx-FLM II was 14.4 +/- 9.9 mg/g (surfactant to albumin) and remained constant across the gestational ages. CONCLUSION: The mean weekly increment of TDx-FLM II is 14.4 +/- 9.9 mg/g and is constant during the latter part of pregnancy. This information, combined with the gestational age, should be useful in treating women with an initial immature test. PMID- 16260212 TI - Maternal plasma concentrations of IGF-1, IGFBP-1, and C-peptide in early pregnancy and subsequent risk of gestational diabetes mellitus. AB - OBJECTIVE: Insulin-like growth factor-1 (IGF-1) and IGF binding protein-1 (IGFBP 1) may be important determinants of glucose homeostasis. We examined the association between circulating concentrations of IGF-1, IGFBP-1 in early pregnancy and development of gestational diabetes mellitus (GDM). STUDY DESIGN: Maternal plasma (collected at 13 weeks) IGF-1, IGFBP-1, and C-peptide were measured using immunoassay. Relative risks (RR) and 95% CIs were calculated. RESULTS: The percentage of the cohort that developed GDM was 5.8% (n = 804). Free IGF-1 and IGFBP-1 were inversely associated with GDM risk, while C-peptide was positively associated with GDM risk (P for trend test < .05). Women with free IGF 1 > or = 1.08 ng/mL experienced a 69% reduced risk of GDM (CI 0.12-0.75) compared with women having concentrations < 0.80 ng/mL. There was a 57% reduced risk of GDM among women with IGFBP-1 > or = 68.64 ng/mL (RR = 0.43, CI 0.18-1.05). Women with C-peptide > or = 3.00 ng/mL experienced a 2.28-fold increased risk of GDM (CI 1.00-5.19) compared with women who had concentrations < 1.45 ng/mL. CONCLUSION: These associations may help to further elucidate the pathologic process of GDM. PMID- 16260213 TI - Protein Z in patients with pregnancy complications. AB - OBJECTIVE: This study was undertaken to evaluate the association between protein Z concentration and pregnancy complications. STUDY DESIGN: A prospective case control study was conducted over a 2-year period to evaluate the prevalence of protein Z deficiency in pregnancy complications. Protein Z levels were measured at the time of diagnosis of complications such as preeclampsia, intrauterine growth restriction, and intrauterine fetal demise. Protein Z deficiency was defined as a plasma level below 1.2 mg/L. In addition to patients presenting with pregnancy complications, healthy age-matched nonpregnant and pregnant women were invited to participate. RESULTS: A total of 145 women were included in the study: 50 nonpregnant women, 34 healthy pregnant women, 29 women with preeclampsia, 25 women presented with intrauterine growth restriction, and 7 women with intrauterine fetal demise. The median protein Z level was similar in healthy pregnant and nonpregnant women (1.63 [0.47-3.1] mg/L and 1.69 [0.7-3] mg/L, respectively). Three women with normal pregnancies had a low protein Z level (8.8%), compared with 8 patients presenting with intrauterine growth restriction (33.3%) and 8 patients with intrauterine fetal demise (50%). Compared with normal pregnancy, the frequency of decreased protein Z was significantly higher in cases of intrauterine growth restriction and in intrauterine fetal demise (relative risk [RR] 1.96, 95% CI 1.16-3.32; P = .041 and RR 3.36, 95% CI 1.65-6.8; P = .0031, respectively), but not in preeclampsia (RR 1.6, 95% CI 0.9-2.8; P = .23). Placenta histologic examination revealed vascular lesions in 50% of patients with protein Z deficiency and in 33% of patients with normal levels of protein Z (RR 0.84; 95% CI 0.6-1.2). CONCLUSION: Protein Z deficiency is associated with late fetal demise and intrauterine growth restriction. The pathophysiologic role of protein Z deficiency, either congenital or caused by the presence of specific antibodies remains unclear and should be further investigated. PMID- 16260215 TI - Embryogenesis of fused umbilical arteries in human embryos. AB - OBJECTIVE: The objective of this study was to elucidate the embryologic basis of fused umbilical arteries in the human. STUDY DESIGN: Twenty-nine human embryo specimens at Carnegie stages 11 through 15 (4-5 weeks after fertilization) were examined histologically, with special reference to the development of umbilical arteries. RESULTS: All embryos at Carnegie stage 11 and 12 had fused umbilical arteries, and 66% of Carnegie stage 13 embryos and 29% of Carnegie stage 14 embryos still had the condition. None of the embryos at Carnegie stage 15 or older had fused umbilical arteries, but there were always 2 arteries present in their umbilical cords. CONCLUSION: Our data suggest that (1) a single umbilical artery splits into 2 as the developmental stage of the embryo advances, (2) that fused umbilical arteries represent a remnant of the embryonic phenotype, and (3) that fused umbilical arteries are embryologically distinct from true single umbilical artery. PMID- 16260214 TI - A free radical scavenger, edaravone, inhibits lipid peroxidation and the production of nitric oxide in hypoxic-ischemic brain damage of neonatal rats. AB - OBJECTIVE: The purpose of this study was to elucidate a role for edaravone, a free radical scavenger 3-methyl-1-phenyl-2-pyrazolin-5-one, in neonatal hypoxic ischemic brain damage. We determined the level of thiobarbituric acid reactive substances as an index of lipid peroxidation and nitric oxide metabolites as nitric oxide production. STUDY DESIGN: Seven-day-old Wistar rats were subjected to left common carotid artery ligation followed by 2 hours of 8% oxygen exposure. Then, the rats were administered edaravone (9 mg/kg) or saline solution intraperitoneally. Cerebrospinal fluid was withdrawn just before the rats were killed at 2, 5, 24, and 48 hours after hypoxia, and brains were removed. The thiobarbituric acid reactive substances and nitric oxide metabolites levels were measured in the brain tissue and cerebrospinal fluid, respectively. RESULTS: On the ligated side, edaravone significantly decreased thiobarbituric acid reactive substances levels at 5 and 24 hours after hypoxia, compared with saline group (P < .01). Edaravone significantly decreased the nitric oxide metabolites level in the cerebrospinal fluid only at 5 hours, compared with saline group (P < .01). CONCLUSION: Edaravone potently and transiently inhibited lipid peroxidation and the production of nitric oxide in the neonatal rat brain after hypoxic-ischemic insult. PMID- 16260217 TI - Human preterm amnion cells cultured in 3-dimensional collagen I and fibrin matrices for tissue engineering purposes. AB - OBJECTIVE: In this study, human preterm amnion cells were investigated in 3 dimensional (3D) cell-matrix culture systems in an attempt to design therapeutic strategies for preterm premature rupture of the membranes. STUDY DESIGN: Three dimensional collagen I and fibrin cell-containing biomatrices were created to mimic the architecture of native amnion. Amnion mesenchymal cells were embedded in 3D matrices, and epithelial cells were placed on top of these matrices. Cell viability and morphology were visualized by DiI-ac-LDL, F-actin, and nuclear staining. Proteolytic activity of matrix metalloproteinases (MMPs) was investigated using gelatine zymography. RESULTS: Preterm amnion epithelial and mesenchymal cells cultured in collagen I and fibrin matrices assume cell morphologies similar to those observed in vivo. Mesenchymal cells were capable of remodelling collagen I, as seen by extensive volume contraction, by 40% at day 1 and 80% at day 5. Matrix contraction was independent of the presence of epithelial cells, and could not be inhibited by GM6001 and/or aprotinin. No contraction was observed in fibrin matrices over 8 days. The migratory response of mesenchymal cells cultured in 3D fibrin matrices supplemented with fibronectin was associated with specific activated MMP-9. CONCLUSION: Three-dimensional fibrin matrices might be useful in amnion cell tissue engineering, including cell matrix transplantation. PMID- 16260216 TI - Insulin and fatty acids regulate the expression of the fat droplet-associated protein adipophilin in primary human trophoblasts. AB - OBJECTIVE: This study was undertaken to test the hypothesis that insulin and fatty acids regulate adipophilin expression in cultured human trophoblasts. STUDY DESIGN: Cytotrophoblasts isolated from term human placentas were cultured in the absence or presence of insulin (10 nmol/L), and a mix of oleic and linoleic acid in serum-free medium. The expression of adipophilin as well as the fatty acid transport proteins (FATP) 2, 3, 4 and 6 was examined. Fat accumulation was quantified by BODIPY staining and fat uptake determined using [3H]-oleic acid. RESULTS: A combination of insulin and fatty acids enhanced the expression of adipophilin (2.3-fold, P < .05). In contrast, the expression of FATPs was unchanged. Furthermore, insulin and fatty acids increased the accumulation of fat droplets in trophoblasts by 4- to 5-fold (P < .05), but had no effect on oleic acid uptake. CONCLUSION: Insulin and fatty acids enhance the expression of adipophilin and the formation of fatty acid droplets in term human trophoblasts. PMID- 16260218 TI - The American Association of Obstetricians and Gynecologists Foundation Scholars Program: additional data on research-related outcomes. AB - OBJECTIVE: The purpose of this study was to ascertain the progress of recipients of research training in obstetrics and gynecology in establishing an active research career in academic medicine. STUDY DESIGN: Existing data were used to examine the extent to which 41 individuals who had received American Association of Obstetricians and Gynecologists Foundation (AAOGF) fellowships had achieved outcomes indicative of a career in academic medicine. Outcomes included employment as a full-time faculty member, receipt of NIH research funding, number of publications, the types of journals in which these articles had appeared, and the type of research (eg, basic vs patient-oriented). RESULTS: Among individuals who were awarded their fellowship between 1984 and 1997, 88% held faculty appointments, and 40% of these positions were in institutions that were more research-intensive that the medical degree-granting institutions of fellows. Slightly more than half of former fellows had successfully competed for NIH research funding, with 22% being awarded at least one R01 grant. Overall, fellows produced a total of 878 articles, one third of which appeared in clinical journals, 18% were in basic biomedical research journals, and 48% were in journals that published both types of research. CONCLUSION: Previous AAOGF scholars have actively pursued research careers in academic obstetrics and gynecology. Their performance compares favorably with those of individuals receiving research training in other clinical specialties. A more complete understanding of their performance and the value added by the program would be possible if a core set of data on outcomes were available from other types of training efforts in both obstetrics and gynecology and other relevant disciplines. PMID- 16260219 TI - Ischiorectal abscess after sacrospinous ligament suspension. AB - An ischiorectal abscess in a 66-year-old patient was determined to be an uncommon complication of sacrospinous fixation. The abscess was diagnosed 9 months after the patient had a sacrospinous ligament suspension. She was treated successfully with perianal incision, drainage, and intravenous antibiotics. PMID- 16260220 TI - Application of the three-dimensional maximum mode in prenatal diagnosis of Apert syndrome. AB - Apert syndrome is a rare disorder characterized by coronal craniosynostosis, syndactyly, brachycephaly, midfacial hypoplasia, and central nervous system anomalies, among other malformations. We present a case of Apert syndrome examined at 22 + 0 weeks' gestation. Three-dimensional maximum mode was decisive for the correct prenatal diagnosis by demonstrating the cranial deformities. PMID- 16260221 TI - Placement of a temporary vena cava filter during labor. AB - Placement of a vena cava filter for the prevention of pulmonary thromboembolism in select patients is a well established procedure in critical care medicine. We describe a case of placement and removal of a new removable vena cava filter in a pregnant patient, in this case during early labor. Vaginal delivery was accomplished without incident. PMID- 16260222 TI - Maternal death caused by midgut volvulus after bariatric surgery. AB - A 31-year-old woman with a history of laparoscopic Roux-en-Y gastric bypass surgery presented at 25 6/7 weeks' gestation with complaints of abdominal pain. Maternal death followed midgut volvulus, perforation, and septic shock. PMID- 16260223 TI - Vulvar cellular angiofibroma: a case report. AB - Cellular angiofibroma is a benign growth initially described in 1997, with few reports to date. A 31-year-old woman presented with a 3-year history of a small left labial mass, which had recently increased in size to 5 cm, and was clinically thought to be a lipoma. A simple excision was performed. Histologically, the mass was consistent with a cellular angiofibroma. Ten months later, the growth has not recurred. Cellular angiofibroma is a rare, benign mesenchymal lesion typically occurring on the vulva, and should be considered in the differential diagnosis of a painless, soft, vulvar mass. PMID- 16260224 TI - A tale of 2 pedunculated myomas. AB - We present 2 unusual cases of prolapsed pedunculated submucous myomas. In 1 patient, the prolapsed part measured 12 cm, with a 64-cm intrauterine part. The second patient had prolapsed pedunculated submucous myoma, which subsequently retracted into the uterus. Gynecologists should be aware of unusual presentations of pedunculated submucous myoma to plan surgery. PMID- 16260225 TI - Temporary balloon occlusion of the common iliac artery: new approach to bleeding control during cesarean hysterectomy for placenta percreta. AB - A case of placenta percreta was referred at 31 weeks' gestation. We performed a cesarean hysterectomy preceded by placement of occlusive balloon catheters at bilateral common iliac arteries at 34 weeks' gestation. This simple and safe technique provides satisfactory efficacy for control of profuse bleeding during operation, with blood loss estimated at 800 mL. PMID- 16260226 TI - Episiotomy dehiscence that required intestinal diversion. AB - Postpartum episiotomy dehiscence is a rare complication of vaginal delivery. Forceps-assisted vaginal delivery over mediolateral episiotomy was complicated by infection and dehiscence with rectal injury. A diverting ileostomy was used to permit healing. Episiotomy infection requires early recognition and thorough evaluation to exclude occult rectal injury. PMID- 16260227 TI - Endometriotic umbilical port site metastasis after laparoscopy. AB - We present the case of a 40-year-old woman who had been previously operated for endometrioma 2 years ago by laparoscopy, with the complaint of an umbilical mass with cyclical pain pattern. The dark-colored mass was excised and pathology report revealed it to be an endometriotic implant. PMID- 16260228 TI - On the sites of the negative and positive feedback actions of estradiol in the control of gonadotropin secretion in the rhesus monkey. 1978. PMID- 16260229 TI - Discovery of the hypothalamic gonadotropin-releasing hormone pulse generator and of its physiologic significance. 1992. PMID- 16260231 TI - Steadfastly forward. PMID- 16260233 TI - Medical students self-reported work hours: perception versus reality. AB - OBJECTIVE: The objective of this study was to compare the students' actual work hours with their self-reported work hours during the obstetrics and gynecology clerkship, and to determine whether the number of hours worked correlate with the amount of "scut" reported or students' rating of the quality of the clerkship. STUDY DESIGN: Students self-reported work hours were compared against their actual scheduled hours over 2 different academic years. Pearson's correlation was performed to correlate the actual hours with the amount of reported "scut" work and the overall rating of the quality of the clerkship. RESULTS: The actual hours per week worked by students averaged 59 hours in 2003 and 48 hours in 2004. Students overestimated their work hours both years. Students who worked more hours rated the clerkship lower and the quality of the clerkship significantly improved from 2003 to 2004 (4.2 vs 3.8 P < .03). CONCLUSION: The majority of third-year students overestimate their work hours in obstetrics and gynecology. The rating of the overall quality of the clerkship increase significantly with fewer hours worked, and it is not affected by the amount of "scut" work. PMID- 16260232 TI - The essential elements of undergraduate medical education in obstetrics and gynecology: a comparison of the Association of Professors of Gynecology and Obstetrics Medical Student Educational Objectives and the National Board of Medical Examiners Subject Examination. AB - OBJECTIVE: The objective of this study was to investigate whether the essential elements of the Association of Professors of Gynecology and Obstetrics (APGO) Medical Student Educational Objectives were adequately represented on the National Board of Medical Examiners (NBME) obstetrics and gynecology subject examination, and that the topics questioned on that examination were covered by the APGO objectives. STUDY DESIGN: The Undergraduate Medical Education Committee of APGO and the NBME staff separately reviewed the same 2 NBME obstetrics and gynecology subject examinations. The questions were mapped to the 15 essential elements of the APGO educational objectives and comparisons were made to check how well they matched. RESULTS: All the essential elements of the educational objectives were covered by the NBME subject examination. Of the questions on the examination, 99% were deemed appropriate for medical students with 70% of the questions mapping to "Priority 1" objectives. CONCLUSION: The NBME examination provides an appropriate assessment of mastery of what a medical student should learn, as represented by the APGO Medical Student Educational Objectives. PMID- 16260234 TI - Development and assessment of a Web-based evaluation and management coding curriculum for residents. AB - OBJECTIVE: The purpose was to assess a Web-based learning module for residents on evaluation and management coding, and to determine whether using a point system based on reimbursement is an effective way to measure improvement in performance. STUDY DESIGN: Nineteen residents at a university-based residency completed an online module on evaluation and management coding. Pretest and posttest were administered consisting of 10 actual patient notes from which to abstract the level of service. Tests were scored by percent correct as well as assigning points to correct responses equal to 2004 Medicare reimbursement for that item's level of service. Incorrect responses were debited points equal to the absolute difference in reimbursement between the incorrect and correct evaluation and management level. RESULTS: Average percent correct was 44.2 on the pretest and 45.3 on the posttest (P = 1.0), with only 47% of subjects improving their performance. The mean point score was 513.44 on the pretest and 555.87 on the posttest (P = .02), with 84% improving. Most subjects (84%) rated the module equal or superior to a didactic format and felt that their knowledge was improved by the exercise. CONCLUSION: An electronic learning module on evaluation and management coding for residents, using scoring based on reimbursement schedules, is effective for instruction and assessment and well accepted by learners. PMID- 16260235 TI - Teaching residents coding and documentation: effectiveness of a problem-oriented approach. AB - OBJECTIVE: We sought to assess the effectiveness of a problem-oriented approach to teaching residents accurate coding and documentation of ambulatory gynecology visits. STUDY DESIGN: This was a pilot before-and-after study. Nine resident volunteers underwent 4 individual instructional sessions on coding and documentation with a trained faculty member over 6 weeks. Outcomes were assessed by comparing the appropriateness of procedure and diagnostic codes billed in participant continuity clinic prior to and in the 6 to 9 months following the intervention. RESULTS: Following the intervention, participants demonstrated an increase in the accuracy of coding the correct category of the evaluation and management service, an increase in the appropriate use of modifiers, and a decline in undercoding errors. CONCLUSION: Problem-oriented interactive learning appears to be an effective method of teaching residents proper coding and documentation. PMID- 16260237 TI - Obstetrics-Gynecology resident satisfaction. AB - OBJECTIVE: The objective of the study was to identify factors associated with satisfaction in the Indiana University School of Medicine Obstetrics-Gynecology residency program and the residents' and faculty's perception of whether these factors were extant in our program. STUDY DESIGN: Residents and faculty at the Indiana University School of Medicine Obstetrics-Gynecology program were surveyed using an instrument based on prior primary care specialty investigations. Multivariate regression evaluated the impact of various factors on resident satisfaction. RESULTS: Seventy-seven percent (44 of 57) and 100% (35 of 35) of faculty and residents, respectively, completed the survey. Relevant training, collegiality, adequate resources, workload, care continuity, supportive coworkers, learning environment, autonomy, role ambiguity, and supportive faculty were significantly associated with resident satisfaction. Care continuity, role ambiguity, and learning environment were the areas of largest faculty/resident disagreement. CONCLUSION: Relevant training and collegiality were most strongly linked to resident satisfaction. Three areas of dissatisfaction were identified, and we will seek to remedy these areas. PMID- 16260236 TI - Implementation and evaluation of a genetics curriculum to improve obstetrician gynecologist residents' knowledge and skills in genetic diagnosis and counseling. AB - OBJECTIVE: This study was undertaken to develop, implement, and evaluate a genetics curriculum for obstetrician-gynecologist residents. STUDY DESIGN: We prospectively evaluated the effect of a genetics curriculum on obstetrician gynecologist residents' knowledge and skills. Residents completed a needs assessment and pretest. Educational intervention included 2 3-hour didactic sessions with 1 hour of lecture followed by case discussion and 1 3-hour session of experiential learning using standardized patients who evaluated residents' knowledge and skills in taking family history, drawing genetic pedigrees, and counseling patients. Posttest scores were compared with pretest scores. RESULTS: Needs assessment was completed by all 40 obstetrics and gynecology residents and identified limited and variable genetics education in medical school. Twenty eight of 40 residents attended the entire educational intervention and completed the pretest and posttest, and 25 of 28 showed improved test scores. Residents stated that they were more confident in their ability to take a family history, record a 3-generation pedigree, and counsel patients about genetic conditions after completion of the genetics curriculum. CONCLUSION: This multifaceted genetics curriculum improved residents' knowledge of genetics as well as their confidence in applying genetic concepts as assessed by the pretest and posttest and by their comments in the debrief session. PMID- 16260238 TI - Residency attrition rate in obstetrics and gynecology: are we losing more postgraduates today? AB - OBJECTIVE: The purpose of this descriptive study was to determine the attrition rate in 2003 and to establish where residents matriculate after leaving an obstetrics and gynecology residency program. STUDY DESIGN: A questionnaire was sent by e-mail to all program directors in obstetrics and gynecology residencies in the United States. The questionnaire asked for the number of residents who had left a program, what year of training the resident was in, and whether the departure was a transfer, withdrawal, or dismissal. It asked whether a transfer was to an obstetrics and gynecology residency program or to another specialty; if the resident transferred to another specialty, which specialty did the resident choose. RESULTS: Two hundred nineteen of 253 programs responded (86.5%). Of residents who left programs, 49% left in the first year of training; 34% left in the second year of training; 13% left in the third year of training, and 4% left in the fourth year of training. The reason for attrition was that 75% of the residents transferred to another residency program; 16% of the residents withdrew from training, and 8% of the residents were dismissed. Of the transferring residents, 60% remained in obstetrics and gynecology. CONCLUSION: Although resident attrition was higher than in 1992, more residents remained in obstetrics and gynecology. PMID- 16260239 TI - Personality type and clinical evaluations in an obstetrics/gynecology medical student clerkship. AB - OBJECTIVE: The purpose of this study was to determine whether the Myers-Briggs type inventory extraversion is associated with clinical evaluation ratings that students earn during their (obstetrics/gynecology) junior medical student clerkship. STUDY DESIGN: The Myers-Briggs type inventory was administered to medical students during their obstetrics/gynec clerkship. Bivariate correlations between clinical evaluations, National Board of Medical Examiners subject scores, and data from the Myers-Briggs type inventory extraversion scale were analyzed. RESULTS: Pearson product-moment correlation between clinical and National Board of Medical Examiners subject scores was not significant (r = .25; P = .05). The National Board of Medical Examiners did not show significant correlations with the Myers-Briggs type inventory extraversion data. The clinical evaluations showed a significant correlation (r = .35; P = .005) with Myers-Briggs type inventory extraversion. CONCLUSION: Results show that Myers-Briggs type inventory extraversion is correlated positively with clinical evaluations. The National Board of Medical Examiners subject examination and clinical evaluations were not correlated significantly. Findings question whether clinical evaluation data should be included in the obstetrics/gynecology medical student evaluation process. PMID- 16260241 TI - Self-assessment of resident surgical skills: is it feasible? AB - OBJECTIVE: The purpose of this study was to examine obstetrics and gynecology residents' self-assessment of proficiency on a variety of surgical bench procedures and to compare their ratings with those ratings of trained faculty observers who used instruments that have been shown to be reliable and valid. STUDY DESIGN: As part of a 6-station Objective Structured Assessment of Technical Skills, 74 residents at 5 institutions estimated their overall open and laparoscopic skill level before the testing. After completing each station, residents evaluated their overall and global skills performance. RESULTS: Residents rated their proficiency higher on open skills than on laparoscopic skills. Task-specific, overall, and global assessments were correlated significantly with the faculty ratings (P < .001). Residents tended to rate themselves lower than did faculty on almost all measures; even those residents with poor skills indicated that they were aware of their deficiencies. Overall and global self-assessments increased with each resident level, which indicated good construct validity. CONCLUSION: Residents can rate their overall open and laparoscopic skills, task-specific performance, and global skills with good reliability and validity. Although they tended to score themselves lower than did faculty observers, the correlations are high (ie, residents who give themselves a higher score tended to receive a higher score from faculty, and vice versa). One of the concerns about self-assessment is that residents with poor skills might not be aware of their deficiencies. We did not find that to be the case. Therefore, when residents work on self-directed exercises, task-specific and global checklists can be used for both learning and self-assessment. PMID- 16260240 TI - Focused assessment of surgical performance: difficulty with faculty compliance. AB - OBJECTIVE: This study evaluated faculty compliance in the use of the global surgical rating scale of the Objective Structured Assessment of Technical Skills to rate resident surgical performance after every endoscopic procedure. STUDY DESIGN: For this prospective cohort study, 4 faculty members in the Minimally Invasive Gynecology Surgery Program were asked to rate resident surgical performance using the Objective Structured Assessment of Technical Skills instrument after every case. Faculty compliance was analyzed with respect to the influence of the resident or surgical case characteristics. Faculty and residents completed surveys about the value of the case-by-case ratings. RESULTS: Faculty members used the Objective Structured Assessment of Technical Skills instrument 36% of the time (range, 26%-60%). Faculty member compliance did not vary according to resident or surgical case characteristics. Faculty members did not think the forms had much impact on whether they gave feedback. Residents thought the opportunity to read their ratings was helpful. CONCLUSION: Faculty member compliance with case-by-case surgical performance evaluation of the residents was low. PMID- 16260242 TI - Resident job satisfaction: one year of duty hours. AB - OBJECTIVE: This study was undertaken to assess job satisfaction and quality of life aspects among residents in obstetrics/gynecology before and after the implementation of duty-hour requirements. STUDY DESIGN: We administered a survey to residents before and after duty-hour restrictions, addressing satisfaction with residency training, quality of life, and predictions/impressions of the effect of reduced work hours. RESULTS: Satisfaction with overall residency training as a discrete survey item did not change; however, the composite score from all responses to specific items increased. Several specific clinical and academic items garnered higher satisfaction scores in 2004. Residents reported less-than-anticipated increases in healthiness of their lifestyle and a decrease in interest in teaching. CONCLUSION: This is a prospective assessment of the effect of duty-hour requirements, improvements in residents' perception of their time and ability to study and pursue research and in clinical areas are encouraging. The perception that there is less interest in teaching is of concern. PMID- 16260243 TI - The influence of an audience response system on knowledge retention: an application to resident education. AB - OBJECTIVE: The purpose of the study was to compare delivery methods of lecture material regarding contraceptive options by either traditional or interactive lecture style with the use of an audience response system with obstetrics and gynecology residents. STUDY DESIGN: A prospective, randomized controlled trial that included 17 obstetrics and gynecology residents was conducted. Group differences and comparison of pre/posttest scores to evaluate efficacy of lecture styles were performed with the Student t test. Each participant completed an evaluation to assess usefulness of the audience response system. RESULTS: Residents who received audience response system interactive lectures showed a 21% improvement between pretest and posttest scores; residents who received the standard lecture demonstrated a 2% improvement (P = .018). The evaluation survey showed that 82% of residents thought that the audience response system was a helpful learning aid. CONCLUSION: The results of this randomized controlled trial demonstrate the effectiveness of audience response system for knowledge retention, which suggests that it may be an efficient teaching tool for residency education. PMID- 16260244 TI - Obstetrics and gynecology residents as teachers of medical students: predictors of excellence. AB - OBJECTIVE: The purpose of this study was to assess variables that might predict which intern candidates will become excellent teachers of medical students. STUDY DESIGN: This retrospective cohort study compared demographic characteristics, previous work experience, United States Medical Licensing Examinations scores, honors on core clerkships, membership in Alpha Omega Alpha, and match list ranking of 43 residents to identify predictors of excellent teaching evaluations during residency. RESULTS: Fifteen residents (35%) were identified as excellent teachers. They were more likely to have had previous work experience, to be older, or to be male. They were not more likely to have higher United States Medical Licensing Examinations test scores, more honors grades, Alpha Omega Alpha membership, or a higher rank list position. CONCLUSION: Work experience, age, and male gender are associated with increased likelihood of being identified as an excellent teacher by medical students. Programs in which residents have a significant role as teachers of students may consider these factors in the residency selection process. PMID- 16260246 TI - Candid candidate comments: the relationship between residency program selection factors and match list placements from ranked applicants. AB - OBJECTIVE: The purpose of this study was to explore the relationship between residency program selection factors and placements we received on candidates' match lists. STUDY DESIGN: An annual (2001-2004) telephone survey of applicants (n = 140) ranked for acceptance to the department's residency program addressed 16 program selection factors important in choosing our site. Candidates' comments about these program selection factors were recorded. Logistic regression was conducted with program selection factors as independent variables and program receipt of candidates' top quartile match list rankings as the dependent variable. RESULTS: The overall response rate was 68% (n = 95). Applicants' positive ratings on 3 program selection factors were related to our receipt of top quartile match list rankings: 1) our capacity to meet personal career goals (odds ratio [OR] 4.59); 2) positive faculty-resident relationships (OR 1.97); and 3) suitable location (OR 1.96). CONCLUSION: The single most important factor contributing to receipt of candidates' top quartile match list placements was positive perceptions of our program's abilities to meet their personal career goal needs. PMID- 16260245 TI - A multicenter study to determine motivating factors for residents pursuing obstetrics and gynecology. AB - OBJECTIVE: This study was undertaken to determine why residents choose obstetrics and gynecology. STUDY DESIGN: Applicants to obstetrics and gynecology residency programs were surveyed; a 5-point scale (5 = most important) was used to rate various aspects of the specialty. Univariate statistics were performed. Bivariate analysis comparing results that were based on gender and timing of decisions was completed with Student t test, chi2, and Kruskal-Wallis tests. RESULTS: A total of 153 applicants (42% response rate) from 10 programs participated; 85.3% of respondents were female. Surgical opportunities, variety of clinical experience, and fast-paced/high-acuity experiences attract applicants to obstetrics and gynecology. When considering programs, resident camaraderie, gynecologic experience, and commitment to education were most important. Over 70% of residents decided to pursue obstetrics and gynecology during or after their third year clerkship. CONCLUSION: Surgical opportunities and clinical variety appeal to applicants. The majority choose obstetrics and gynecology during or after their core clerkship. In addition, program dynamics are important when choosing a residency. PMID- 16260247 TI - Educational games in an obstetrics and gynecology core curriculum. AB - OBJECTIVE: The purpose of this study was to compare learning satisfaction and effectiveness using traditional lecture or educational game in teaching medical students about ectopic pregnancy. STUDY DESIGN: Third-year medical students were randomized to instruction about ectopic pregnancy through either standard lecture or educational Jeopardy style game. Students in each group completed a pretest, posttest, and satisfaction survey. Experts in ectopic pregnancy validated the pretest and posttest. The satisfaction survey was taken from published validated tests. Paired samples t test was used to compare pretest and posttest scores. Independent samples t tests were used to compare test scores and satisfaction responses between groups. Chi-square tested dichotomous satisfaction responses. RESULTS: All 104 students in both groups showed significant improvement in learning about ectopic pregnancy (P < .001) on pre- and posttest comparison, with scores being almost identical. Students in the group randomized to game format rated it higher in stimulating faculty/student interaction, helping retain information, and overall enjoyment than students participating in the lecture method (P < .001). In addition, students in the game group responded positively that the format was interactive, stimulated their interest, and kept them engaged in class content (P < .001). CONCLUSION: The innovative educational game format was as effective as standard lecture in educating students about ectopic pregnancy, while being more enjoyable and stimulating. Based on these conclusions, we hope to motivate other teachers in obstetrics and gynecology to use innovative teaching methods to provide a more enjoyable, stimulating, and active means of effective medical education. PMID- 16260248 TI - Medical student identification of domestic violence as measured on an objective, standardized clinical examination. AB - OBJECTIVE: This study examined the incidence and predictors of domestic violence screening by third-year medical students at an end of clerkship Objective Standardized Clinical Examination. STUDY DESIGN: Two hundred and seventy-five third-year medical students completed an 8-station end of clerkship Objective Standardized Clinical Examination as part of this retrospective observational study, one with nonspecific abdominal pain and possible domestic violence. Checklists on history, physical, communication, and interpersonal skills were collected. Domestic violence screening was analyzed by logistic regression and analysis of variance. RESULTS: The incidence of domestic violence screening by history alone was 34% before the physical. Interpersonal scores on the overall exam and domestic violence station, but not gender or rotation sequence, predicted domestic violence questioning. CONCLUSION: In this standardized patient study there was a low rate of domestic violence screening by history. PMID- 16260249 TI - A new curriculum for hysteroscopy training as demonstrated by an objective structured assessment of technical skills (OSATS). AB - OBJECTIVE: The purpose of this study was to develop an effective curriculum to teach hysteroscopy, as well as to develop an objective assessment instrument (OSATS). STUDY DESIGN: All 24 residents in our training program attended a 3-hour hysteroscopy curriculum in each of 3 years. Each year after the curriculum, an OSATS was administered consisting of an assembly and operative task. Each task was evaluated with a task-specific checklist and a previously validated global assessment form. Twenty-four residents from an outside institution served as controls. All residents were examined by blinded and unblinded examiners. RESULTS: The reliability coefficient was .7857 for the checklist and .9763 for the global scale. Interrater reliability for the checklist was .7478 and ranged from .4222 to .9329 for the global instruments. Evaluation of construct validity revealed that the junior residents at both locations did significantly worse on the checklist and global scale than the senior residents for all 3 years of the study (P < .001). Those residents provided the curriculum did significantly better on both the checklist and the global scale than the resident controls with a mean of 45% versus 63% for the checklist and 18.3 versus 24.9 for the global score. (P = .001 checklist, P = .007 global scale). CONCLUSION: This curriculum is an effective way to impart knowledge and skill in the assembly and use of the operative hysteroscope. The checklist developed for this OSATS has excellent reliability and construct validity. PMID- 16260250 TI - Measurement of endometrial stripe thickness by obstetrics and gynecology residents. AB - OBJECTIVE: How well do obstetrics and gynecology residents measure the endometrial stripe by transvaginal sonography? STUDY DESIGN: Three obstetrics and gynecology residents at the University of Texas Medical Branch from each year level were tested for their ability to perform endometrial stripe measurements. Measurements of endometrial stripe thickness within 1 mm of the reference value that was obtained by an expert sonographer were deemed to be correct. RESULTS: Residents correctly measured the endometrial stripe in 14 of 24 cases (58%). Postgraduate year-4 residents correctly measured endometrial stripe thickness in 5 of 6 cases (83%); lower level residents were correct in 9 of 18 cases (50%). The most common error that was noted was an incorrect image plane in 25 of 47 attempts (53%). Fewer errors were committed by upper level as compared with lower level residents (P < .05). CONCLUSION: By the end of residency, residents can accurately measure the endometrial stripe using transvaginal sonography. Emphasis should be placed on the improvement of the measurement skills. PMID- 16260251 TI - Impact of 1996 Residency Review Committee Obstetrics-Gynecology primary care requirements on residency training and surgical procedures. AB - OBJECTIVE: This study evaluated the impact of required primary care rotations in obstetrics-gynecology residency training after 1996. STUDY DESIGN: A questionnaire was sent to the 1994 to 2003 graduates from 1 residency program, and records of surgical procedures completed during residency were analyzed. RESULTS: Thirty-nine of 46 graduates participated in the study (response rate 85%). Required primary care training was associated with increased confidence in providing primary care (81.5% versus 54.5%, P = .12) but less agreement that obstetrics-gynecology is a primary care specialty (21.4% versus 45.5%, P = .23). Abdominal hysterectomies and vaginal hysterectomies per resident did not decrease (127 versus 113, P = 0.149, and 55 versus 48, P = .06, respectively). Adjusted for temporal trends, cesarean sections per resident decreased (366 versus 321, P = .009). CONCLUSION: Residents maintained adequate rates of major inpatient surgical procedures after implementation of required primary care training. There was a tendency for residents who graduated after 1996 to have less favorable attitudes about primary care. PMID- 16260252 TI - Improved performance and student satisfaction after implementation of a problem based preclinical obstetrics and gynecology curriculum. AB - OBJECTIVE: This study was undertaken to assess student performance and satisfaction as a preclinical obstetrics and gynecology course changed from a didactic to a problem-based format. STUDY DESIGN: We prospectively compared examination scores and course evaluations given to 162 second-year medical students over 4 years: 2 years before and 2 years after the curricular change, to assess student performance and satisfaction with learning. We used analysis of variance for the analysis of student performance and likelihood-ratio test for analysis of student satisfaction. RESULTS: Mean examination scores increased from 79.1% to 84% after implementation of the problem-based curriculum (P < .0001). The data showed statistically significant improvement in satisfaction with course content, learning objectives, learning resources, instructional methods, and course examinations. CONCLUSION: Students' satisfaction and performance in a preclinical obstetrics and gynecology course improved significantly when problem based learning methods were introduced. PMID- 16260253 TI - Risk assessment for neonatal respiratory distress syndrome with FLM II combined with gestational age. PMID- 16260257 TI - Evidence does not support cervical preservation. PMID- 16260255 TI - Lymphatic mapping for endometrial cancer: is hysteroscopic injection a safe technique for sentinel lymph node biopsy? PMID- 16260259 TI - Fetal monitoring with the ST analyser: need for a long-term follow-up of the infants. PMID- 16260261 TI - What makes words sound similar? AB - Although similarity plays an important role in accounts of language processing, there are surprisingly few direct empirical studies of the phonological similarity between words, and it is therefore not clear whether similarity comparisons between words involve processes similar to those involved in other cognitive domains. In five experiments, participants chose which of two monosyllabic pseudo-words sounded more similar to a target pseudo-word. Our results are generally consistent with the structural alignment theory of comparisons between complex mental representations, suggesting that phonological word similarity parallels similarity involving other kinds of information including visual objects and scenes, events, and word meanings. We use our results to test new metrics of word similarity, and identify predictions for future similarity research both in the domain of word sounds and in other cognitive domains. PMID- 16260262 TI - Different time courses of integrative semantic processing for plural and singular nouns: implications for theories of sentence processing. AB - The research investigated the time course of integrative semantic processing during sentence processing. Reading time was measured on sentences containing an NP composed of an adjective and a noun whose combined meaning was plausible or anomalous (Experiment 1) or was typical or atypical (Experiment 2). The noun in the NP was either plural or singular. Plural nouns were expected to be more rapidly integrated with a preceding adjective than singular nouns because plural nouns can be ruled out as the first noun in a noun compound more rapidly than singular nouns. The results confirmed this prediction, showing that the effects of semantic plausibility and typicality were observed immediately during the processing of plural nouns, but were observed at a delay following the processing of singular nouns. Implications for theories of sentence processing are discussed. PMID- 16260263 TI - On the limits of infants' quantification of small object arrays. AB - Recent work suggests that infants rely on mechanisms of object-based attention and short-term memory to represent small numbers of objects. Such work shows that infants discriminate arrays containing 1, 2, or 3 objects, but fail with arrays greater than 3 [Feigenson, L., & Carey, S. (2003). Tracking individuals via object-files: Evidence from infants' manual search. Developmental Science, 6, 568 584; Feigenson, L., Carey, S., & Hauser, M. (2002). The representations underlying infants' choice of more: Object files versus analog magnitudes. Psychological Science, 13(2), 150-156]. However, little is known about how infants represent arrays exceeding the 3-item limit of parallel representation. We explored possible formats by which infants might represent a 4-object array. Experiment 1 used a manual search paradigm to show that infants successfully discriminated between arrays of 1 vs. 2, 2 vs. 3, and 1 vs. 3 objects. However, infants failed to discriminate 1 vs. 4 despite the highly discriminable ratio, providing the strongest evidence to date for object-file representations underlying performance in this task. Experiment 2 replicated this dramatic failure to discriminate 1 from 4 in a second paradigm, a cracker choice task. We then showed that infants in the choice task succeeded at choosing the larger quantity with 0 vs. 4 crackers and with 1 small vs. 4 large crackers. These results suggest that while infants failed to represent 4 as "exactly 4", "approximately 4", "3", or as even as "a plurality", they did represent information about the array, including the existence of a cracker or cracker material and the size of the individual objects in the array. PMID- 16260264 TI - Rhesus monkeys (Macaca mulatta) spontaneously compute addition operations over large numbers. AB - Mathematics is a uniquely human capacity. Studies of animals and human infants reveal, however, that this capacity builds on language-independent mechanisms for quantifying small numbers (<4) precisely and large numbers approximately. It is unclear whether animals and human infants can spontaneously tap mechanisms for quantifying large numbers to compute mathematical operations. Moreover, all available work on addition operations in non-human animals has confounded number with continuous perceptual properties (e.g. volume, contour length) that correlate with number. This study shows that rhesus monkeys spontaneously compute addition operations over large numbers, as opposed to continuous extents, and that the limit on this ability is set by the ratio difference between two numbers as opposed to their absolute difference. PMID- 16260265 TI - Time to get a new mountain? The role of function in children's conceptions of natural kinds. AB - Prior research indicates that young children are promiscuously teleological, attributing purpose not only to artifacts, but also to living and non-living natural entities. This study further examines the role of function in children's reasoning about different object kinds by indirectly probing children's intuitions about what types of entities can be rendered functionless. Specifically, children were asked to decide whether entities that could no longer perform certain activities should be fixed/replaced (e.g. "Do you need to get a new one?"). Results reveal that young children broadly view both artificial and natural kinds that can no longer perform certain activities as needing to be fixed or replaced. These findings suggest that the teleo-functional bias not only influences children's explanatory preferences but also their category judgments. PMID- 16260266 TI - Mesothelioma versus carcinoma: getting easier? PMID- 16260268 TI - Value of estrogen and progesterone receptor immunostaining in distinguishing between peritoneal mesotheliomas and serous carcinomas. AB - The differential diagnosis between peritoneal mesotheliomas and serous carcinomas involving the peritoneum may be difficult, but it can be facilitated by the use of immunohistochemistry. To determine whether estrogen receptors (ER) or progesterone receptors (PR) may have any value as immunohistochemical markers for discriminating between these malignancies, 40 serous carcinomas of the ovary metastatic to the peritoneum, 7 primary peritoneal serous carcinomas, 30 epithelioid peritoneal malignant mesotheliomas, 5 well-differentiated papillary mesotheliomas, and 4 adenomatoid tumors were immunostained for ER and PR. Reactivity for ER was obtained in 35 (88%) of the metastatic serous carcinomas of the ovary and 6 (86%) of the primary peritoneal serous carcinomas, whereas positivity for PR was observed in 24 (60%) of the metastatic serous carcinomas and 4 (56%) of the primary peritoneal serous carcinomas. None of the mesotheliomas or adenomatoid tumors expressed ER or PR. It is concluded that, because of its high sensitivity for serous carcinomas, ER immunostaining could be very useful in distinguishing between serous carcinomas and peritoneal mesotheliomas. Immunostaining for PR, however, has little practical utility. PMID- 16260267 TI - GADD45-alpha expression in cirrhosis and hepatocellular carcinoma: relationship with DNA repair and proliferation. AB - Growth arrest and DNA damage 45-alpha (GADD45-alpha) is a nuclear protein involved in maintenance of genomic stability, DNA repair, and suppression of cell growth through interaction with nuclear elements, including cyclin-dependent kinase inhibitor 1A (CDKN1A) and PCNA. In this study, GADD45-alpha expression was assessed in 28 cases of hepatocellular carcinoma (HCC) and matched cirrhosis tissues, and correlated with the presence of DNA-bound PCNA and CDKN1A as markers of DNA repair, as well as with clinicopathologic variables including histopathologic grade, tumor size, nodularity, viral status, alpha-fetoprotein serum levels, and p53 and Ki67 immunostaining. GADD45-alpha and CDKN1A messenger RNA (mRNA) were analyzed by reverse transcriptase-polymerase chain reaction. GADD45-alpha protein expression was evaluated by Western blot (WB) and enzyme linked immunosorbent assays (ELISAs). PCNA and CDKN1A DNA-bound fractions were determined by WB. GADD45-alpha mRNA was down-regulated in 20 of 26 HCCs with respect to matched cirrhosis, but no correlation was found with the corresponding protein levels assessed by both WB and ELISA. GADD45-alpha and CDKN1A protein levels were related to each other both in cirrhotic and in neoplastic tissues, and a concordant up- or down-regulation was observed in HCCs with respect to cirrhosis. DNA-bound PCNA and CDKN1A were present in 5 HCCs and were associated with higher GADD45-alpha protein levels assessed by ELISA. No significant association was found in HCCs between GADD45-alpha protein expression and histopathologic grading, nodule size, focality, and proliferation, whereas a positive correlation was found with alpha-fetoprotein serum levels. In conclusion, GADD45-alpha mRNA was down-regulated with respect to matched cirrhosis in most HCCs; however, no correlation was found between mRNA and protein levels. GADD45-alpha protein levels were higher in HCCs with DNA-bound CDKN1A and PCNA, suggesting a possible role in DNA repair. PMID- 16260269 TI - The pathology student fellowship program at the University of Vermont: 1956-2005. AB - Student fellowship programs in pathology offer a unique educational experience for selected medical students. In this report, the specialty preferences of former student fellows graduating from the University of Vermont were analyzed. Since 1956, 110 students have participated in this program, of whom 33 chose pathology as a career. These individuals represented 32.6% of all Vermont graduates who entered pathology between 1958 and 2005. In addition, former student fellows were more likely to become academic pathologists and to obtain subspecialty certification. Furthermore, based on comments received from former student fellows, the program had a positive impact on the medical education and career of pathologists and nonpathologists alike. It is concluded that student fellowship programs represent a powerful recruitment tool for pathology generally and for academic pathology in particular. The development of a database to track these programs and their graduates is essential to fully assess the educational impact of pathology student fellowship programs nationally. PMID- 16260270 TI - Number and location of nucleoli and presence of apoptotic bodies in diagnostically challenging cases of prostate adenocarcinoma on needle biopsy. AB - There is limited published data regarding the significance of the number or position of nucleoli and the presence of apoptotic bodies in diagnostically challenging cases of adenocarcinoma of the prostate on needle biopsy material. One hundred consecutive prostate cancers on needle biopsy were sent because of diagnostic difficulty to an expert in urological pathology, and the remaining normal benign prostatic glands on the same core were evaluated for the number and location of nucleoli and for the presence of mitotic figures and apoptotic bodies. The Gleason scores of the cases were 6 (86%), 7 (9%), and 8 to 10 (5%). For comparison, the same parameters were evaluated in mimickers of cancer on needle biopsy from other cases, including partial atrophy (n = 135), fully developed atrophy (n = 89), adenosis (n = 50), prostate glands with acute inflammation (n = 50), and high-grade prostatic intraepithelial neoplasia (n = 100). Findings were recorded under high dry magnification (x40) using hematoxylin and eosin-stained sections. Although the number and position of nucleoli did not discriminate between cancer and benign mimickers, mitotic figures and apoptotic bodies were more commonly seen in cancer. Apoptotic bodies in particular were seen fairly frequently (34%) in prostatic adenocarcinoma (also seen in 13% of high-grade prostatic intraepithelial neoplasia), yet rarely in benign mimickers on needle biopsy. Our findings indicate that the presence of apoptotic bodies should be added to the list of histological features that are helpful in the diagnosis of challenging cases of prostate cancer on needle biopsy. PMID- 16260271 TI - Comparative study for the detection of peritubular capillary C4d deposition in human renal allografts using different methodologies. AB - Detection of peritubular capillary (PTC) C4d deposition in tissue sections of renal allograft biopsies became an important aid in the diagnosis of antibody mediated rejection. Pathologists in many major transplant centers now routinely stain renal allograft biopsies for C4d. Currently, there are 3 commercially available antibodies. Two of these antibodies are monoclonal and are usually used with either a 3- or a 2-step indirect immunofluorescence (IF) methodology on frozen sections. A polyclonal antibody is used on formalin-fixed, paraffin embedded tissue section with an immunoperoxidase detection system. The goal of our study was to compare these antibodies and methodologies in our renal allograft biopsy material. Twenty renal allograft biopsies with diffuse or focal PTC C4d staining, using immunofluorescence methods on frozen sections, were selected for this study. These biopsies were tested with the 3 commercially available anti-C4d antibodies (Biogenesis, Brentwood, Calif, cat no. 222-8004; Quidel Corporation, Santa Clara, Calif, cat no. A213; and ALPCO Diagnostic, Windham, NH, cat no. 004-BI-RC4D). Both monoclonal antibodies (Biogenesis and Quidel) were tested with a 3- and a 2-step indirect IF method on frozen sections. The polyclonal antibody (ALPCO) was applied to formalin-fixed paraffin sections using immunoperoxidase methodology. In selected cases, the polyclonal antibody was tested on frozen sections with a 3-step indirect IF method. To exclude possible false-negative staining with the IF method, we selected 10 additional biopsies that showed PTC margination of inflammatory cells, but were C4d-negative or only focally positive, and tested them with the ALPCO antibody on paraffin sections. We have found that all methodologies and antibodies tested provided adequate results with only minor differences between them. Perhaps the most sensitive method is the 3-step indirect IF on frozen sections using one of the monoclonal antibodies. We prefer the 2-step indirect IF method with the Quidel monoclonal antibody because of its simplicity, quick turnaround time, and relatively low cost. The advantages and disadvantages of the individual methodologies are discussed. PMID- 16260272 TI - Up-regulation of insulin-like growth factor axis components in human primary prostate cancer correlates with tumor grade. AB - There is an evidence that components of the insulin-like growth factor (IGF) signaling pathway are involved in the development and progression of prostate cancer. The aim of the present study was to provide a comprehensive analysis of the expression levels of proteins of the IGF axis in prostate cancer. We studied expression of the ligands IGF-I and IGF-II, the inhibitory IGF binding protein-3, the type I IGF receptor (IGF-IR), and the downstream mediator insulin receptor substrate-1 by immunohistochemistry in 56 tissue specimens (28 low-grade and 28 high-grade prostate adenocarcinomas). Protein expression in tumor areas, prostatic intraepithelial neoplasias (PINs), and adjacent benign prostatic tissue were evaluated regarding staining intensity and fraction of positive cells. An immunoreactivity score was established from staining intensity and fraction of positive cells, and correlated with the prognostic clinicopathologic parameters prostate-specific antigen serum levels, Gleason score, and TNM stage. The expression levels of all proteins investigated, except IGF binding protein-3, were up-regulated in PIN and in cancer. IGF-I and IGF-II expression showed a higher expression in high-grade compared with low-grade tumor areas. IGF-I and IGF-II and insulin receptor substrate-1 immunoreactivity was higher in tumors from patients with preoperative prostate-specific antigen serum levels 10 ng/mL or greater, and IGF-II expression was correlated with Gleason score. The data indicate significant alterations in the IGF system as prostate cancer develops. Differential expression of growth-stimulating components of the IGF system may be associated with the malignant phenotype and more aggressive tumor behavior. Expression of IGFs, especially IGF-II, may be predictors of the outcome of prostate cancer. PMID- 16260273 TI - eIF-4E expression is associated with histopathologic grades in cervical neoplasia. AB - Translation initiation factor eIF-4E (eukaryotic initiation factor 4E) is a 25 kd messenger RNA cap-binding phosphoprotein and is involved in the initiation of protein synthesis. The expression is known to be elevated in several carcinomas as compared with normal tissues and benign lesions. In the present study, we undertook to determine whether eIF-4E expression is associated with progression in cervical neoplasia. eIF-4E expression was evaluated by immunohistochemistry in 88 formalin-fixed, paraffin-embedded cervical tissues; 10 normal cervical specimens; 19 low-grade cervical intraepithelial neoplasias (CINs); 19 high-grade CINs; and 40 invasive squamous cell carcinomas (ISCCs). In addition, eIF-4E expression was evaluated at the RNA level in fresh frozen cervical carcinoma tissues by real-time quantitative reverse transcriptase polymerase chain reaction. Immunohistochemical staining showed that eIF-4E expression was undetectable in most normal cervical squamous epithelial tissues (90%), but variable staining was observed in the basal layer of all normal endocervical glands. eIF-4E expression, which was mainly observed as cytoplasmic staining, gradually increased in accordance with histopathologic grade in the order low grade CIN < high-grade CIN < ISCC (P < .001) and, in particular, was strongly detected in all ISCC cases. Furthermore, real-time quantitative reverse transcriptase polymerase chain reaction revealed that eIF-4E expression in tumor was significantly enhanced versus normal cervical tissues (P = .037). These results suggest that eIF-4E may play a significant role in tumor progression of cervical neoplasia and may represent useful markers for malignant transformation of cervical squamous cells. PMID- 16260274 TI - Smad4 protein expression correlates with grade, stage, and DNA ploidy in prostatic adenocarcinomas. AB - The tumor suppressor gene Smad4 (DPC4) has been localized to chromosome 18q21.1 and is a member of the Smad family that mediates the transforming growth factor beta signaling pathway suppressing epithelial cell growth. However, variable expression of this protein has been reported, with a loss in some cancers and increased expression in others. Given both the variability and lack of consensus reported regarding Smad4 expression in prostate cancer, we assessed Smad4 immunoreactivity in prostatic adenocarcinomas (PACs). Formalin-fixed, paraffin embedded tissue sections from 133 PACs were immunostained by a manual method using indirect biotin streptavidin horseradish peroxidase and diaminobenzidine detection using a monoclonal mouse antihuman Smad4 antibody (sc-7966; Santa Cruz Biotechnology Inc, Santa Cruz, Calif). Nuclear immunoreactivity and cytoplasmic immunoreactivity were each semiquantitatively scored based on intensity and percentage of positive cells. Deoxyribonucelic acid ploidy was determined on Feulgen-stained tissue sections by static image analysis. Results were correlated with morphological and prognostic variables. Variable nuclear and cytoplasmic Smad4 positivity was noted in the adjacent benign glands in all cases. Of 133 PACs, 64 (48%) featured increased nuclear and 68 (51%) featured increased cytoplasmic protein expression. Nuclear Smad4 overexpression correlated with tumor grade (P = .02), stage (P = .04), and DNA ploidy (P = .04). Cytoplasmic overexpression correlated with tumor grade (P = .04) and DNA ploidy (P = .04) while showing a trend for correlation with tumor stage (P = .08). Neither nuclear nor cytoplasmic Smad4 overexpression correlated with postsurgical biochemical disease recurrence. Smad4 protein expression persists in PACs compared with benign glands, with both nuclear and cytoplasmic overexpression correlating with prognostic variables indicative of aggressive tumor behavior. Given the significant reported variability of Smad4 in several different cancers, further studies in prostate and other tumors are warranted to elucidate its role in tumorigenesis. PMID- 16260275 TI - Diagnosis of focal nodular hyperplasia of the liver by needle biopsy. AB - Focal nodular hyperplasia (FNH) of the liver can be a difficult diagnosis to establish in limited diagnostic samples such as a needle-core tissue biopsy, especially for pathologists with limited experience with the lesion. To characterize the features that can be used to make the diagnosis, we reviewed and analyzed the clinicopathologic features of 100 consecutive cases submitted for consultation in which we were confident of the diagnosis of FNH in needle biopsy material. A diagnosis of FNH was correctly made by the contributing pathologist in 24 of the 100 referred cases. Most of the patients (81%) were women of childbearing age with a mean age of 36.75 +/- 9.82 years. Most of the patients (70%) were asymptomatic at diagnosis. The most consistent diagnostic histological feature of FNH in needle biopsy was the presence of ductular reaction with varied intensity at the junction of the fibrous septa with the hepatocellular component, which was present in all 100 cases. Thick abnormal arteries were seen in all but 2 cases (n = 98). Features of chronic cholestasis with cholate stasis and accumulation of copper (demonstrable by the rhodanine stain) and copper-binding protein (demonstrable with the Victoria blue stain) were nearly as common (n = 94). A confident diagnosis of FNH can be made with a needle biopsy, especially if the biopsy is known to come from a mass, and the lesion contains characteristic fibrosis with ductules at the interface between hepatocytes and the fibrous region, prominent arteries, and benign hepatocytes with features of chronic cholestasis. PMID- 16260276 TI - Immunohistochemical expressions of cytokeratins, mucin core proteins, p53, and neuroendocrine cell markers in epithelial neoplasm of appendix. AB - Epithelial neoplasms of appendix are infrequent, and their pathological features are not fully characterized. We collected 33 cases of appendiceal tumors and examined immunohistochemically the expression of cytokeratins (CK, CK7, and CK20), mucin core protein (MUC1, MUC2, MUC5AC, and MUC6), E-cadherin, chromogranin A, and p53 protein. Gene analysis of TP53 was also conducted on exons 5 to 8. Clinically, mucinous tumors were predominant in females. Immunohistochemically, all the tumors expressed CK20, whereas CK7 was positive in one third of the cases. Similarly, MUC2 was expressed in all the tumors, whereas MUC1 and MUC5AC were detected in about a half of the cases. Although chromogranin A-positive cells are generally sparse in normal appendix, they were more common in mucinous tumors than in nonmucinous tumors. Contrary to the previous data reported (Mod Pathol 2002;15:599-605), mucinous carcinoma exhibited a higher frequency of p53-positive cells (mean 29%) compared with mucinous adenoma (2.8%) (P < .001), whereas nonmucinous tumors showed high levels of p53-positive cells to similar extent (51%-67%) in both adenoma and carcinoma. The high expression of p53 protein coincided with the presence of mutations in multiple sites of TP53 gene in mucinous tumors. This is the first report that characterized the immunophenotypic profile of appendiceal epithelial neoplasms with an emphasis of a higher frequency of p53 positivity in mucinous carcinoma cases compared with mucinous adenoma in the appendix. PMID- 16260277 TI - Diagnostic significance of aquaporin-1 in liver tumors. AB - The diagnostic utility of aquaporin (AQP)-1 in liver tumors was tested and compared with other well-established markers. In 30 cholangiocarcinomas (CCs), 20 hepatocellular carcinomas (HCCs), and 10 metastatic colorectal carcinomas (MCCs) of the liver, expression of AQP-1, CD10, cytokeratin (CK) 7, CK20, and polyclonal carcinoembryonic antigen (pCEA) was tested. In addition, staining patterns of CD10 and pCEA were analyzed. To compare the selectivity of AQP-1 and CK7 as possible markers for differentiated cholangiocytes, liver biopsies of cholestatic disease were also analyzed. Aquaporin-1 expression was found in 93% of all CCs compared with 0% of HCC (P < .000001) and with 30% of MCC (P < .01). CD10 was positive in 16.7% of CC compared with 40% of HCC (P < .04) and to 20% of MCC (not significant). Cytokeratin 7 was positive in 90% of CC compared with 10% of HCC (P < .00001) and with 20% of MCC (P < .0001). Cytokeratin 20 was positive in 90% of MCC compared with 16.7% of CC (P < .0001) and with 20% of HCC (P < .00001). Canalicular staining patterns of CD10 and pCEA were observed in HCC (100% and 89.5%, respectively) but typically not in CC (0% and 6.7%, respectively) and never in MCC. In cholestatic disease, AQP-1 was expressed in differentiated epithelial cells of the bile ducts, whereas CK7-positive hepatocytes of Rappaport zone 1 did not show any AQP-1 reactivity. Therefore, AQP-1 seems to be a highly selective marker for differentiated cholangiocytes and can be very helpful in the differential diagnosis of liver tumors. PMID- 16260279 TI - Ovarian serous cystadenoma with associated genital rhabdomyoma. PMID- 16260278 TI - Clonal proliferation of cyclin D1-positive mantle lymphocytes in an asymptomatic patient: an early-stage event in the development or an indolent form of a mantle cell lymphoma? AB - Mantle cell lymphoma (MCL) is a B-cell neoplasm with a relatively aggressive clinical course. There is a very small subgroup of patients who present with atypical lymphocytes in peripheral blood, with or without lymphocytosis, lymphadenopathy, or splenomegaly, and with an indolent clinical course. They frequently show mutated IgV(H) genes and CD5 negativity. We report an asymptomatic elderly patient who presented with a single submandibular lymphadenopathy. The biopsy showed immunophenotype and t(11;14)(q13;q32) consistent with MCL. The abnormal lymphoid population was also detected in peripheral blood and bone marrow. The patient has remained asymptomatic for 5 years without receiving any therapy. It is uncertain whether these cases represent an early-stage event in the development or an indolent form of MCL. The existence of such asymptomatic patients with an indolent clinical course should induce a strict clinical judgment in terms of therapeutic decisions. PMID- 16260280 TI - Comparative genomic hybridization for DNA extracted from cytological smear. PMID- 16260282 TI - Anoxic media design, preparation, and considerations. AB - Exclusion of oxygen from growth media is essential for the growth of anoxic prokaryotes. In general, anaerobic techniques focus on the use of deaerated boiled growth media. Successful enrichment, isolation, and cultivation of anoxic prokaryotes critically depend on the choice of appropriate growth media and incubation conditions. This chapter discusses the requirements of anoxic prokaryotes for growth in the laboratory and different existing methods for their cultivation. PMID- 16260283 TI - Enrichment and isolation of anaerobic hydrocarbon-degrading bacteria. AB - Recent progress in microbiology resulted in the enrichment and isolation of anaerobic bacteria capable of the biodegradation of various hydrocarbons under a variety of electron-accepting conditions. Problems challenging the enrichment and isolation of anaerobic hydrocarbonclastic organisms required new approaches and modifications of conventional microbiological techniques. This chapter summarizes the collective experience accumulated in this area starting from anaerobic sampling precautions and includes all stages of cultivation from the construction of initial incubations to final isolation steps and the evaluation of culture purity. PMID- 16260284 TI - Anammox organisms: enrichment, cultivation, and environmental analysis. AB - Anaerobic ammonium oxidation (anammox) is the microbial oxidation of ammonium with nitrite to dinitrogen gas under strict anoxic conditions mediated by planctomycete-like bacteria. Anammox is not only important in the oceanic nitrogen cycle, but can also contribute substantially to nitrogen removal in municipal and industrial wastewater treatment. This chapter addresses the enrichment and cultivation of anammox bacteria in a sequencing batch reactor (SBR) and a gas lift reactor. The reactors can be operated anoxically as an anammox reactor or as an oxygen-limited "completely autotrophic nitrogen removal over nitrite" (CANON) system. Pure cultures of anammox organisms have not yet been obtained, but anammox cells can be purified to more than 99.5% using a Percoll density gradient centrifugation protocol. Furthermore, we show how anammox communities in natural and man-made ecosystems can be identified and characterized using molecular methods such as fluorescence in situ hybridization (FISH) or Planctomycetes/anammox-specific polymerase chain reaction (PCR). Both techniques are based on retrieved 16S rRNA gene sequences. In addition to 16S rRNA, unique anammox ladderane lipids can also serve as biomarkers to determine the abundance of anammox organisms in environmental samples. PMID- 16260285 TI - Enrichment and isolation of ferric-iron- and humic-acid-reducing bacteria. AB - In anoxic habitats, ferric iron oxides and humic acids are widespread, and ferric iron- and humic-acid-reducing microorganisms presumably play an important role in the oxidation of organic matter. Representative strains of ferric-iron- or humic acid-reducing bacteria were isolated from a wide range of freshwater or marine environments. Most of them are strict anaerobes, and facultatively anaerobic microorganisms reduce ferric iron oxides or humic acids only after oxygen has been consumed. Hence, anaerobic techniques have to be used for the preparation of media as well as for the cultivation of microorganisms. Furthermore, special caution is needed in the preparation of ferric iron oxides and humic acids. PMID- 16260286 TI - Enrichment, cultivation, and detection of reductively dechlorinating bacteria. AB - Strategies and procedures for enriching, isolating, and cultivating reductively dechlorinating bacteria that use chloroorganic compounds as metabolic electron acceptors from environmental samples are described. Further, nucleic acid-based approaches used to detect and quantify dechlorinator (i.e., Dehalococcoides) specific genes are presented. PMID- 16260287 TI - Enrichment and isolation of iron-oxidizing bacteria at neutral pH. AB - Methods are provided for the culture of neutrophilic Fe-oxidizing bacteria (FeOB) that grow under microaerobic and anaerobic conditions. For oxygen-requiring lithotrophic Fe oxidizers, it is essential that both O2 and Fe(II) concentrations are kept low, but that an adequate flux of both O2 and Fe(II) are provided to support growth. Techniques using opposing gradients of Fe(II) and O2 are discussed. Preparation of stock solutions of FeS and FeCl2 are described. Methods for the culture of anaerobic FeOB that utilize nitrate or light (anoxygenic photosynthesis) are discussed and presented briefly. PMID- 16260288 TI - High-throughput cultivation of microorganisms using microcapsules. AB - This chapter describes a universal and novel method that provides access to the immense reservoir of untapped microbial diversity by cultivation. This technique uses microcapsules to encapsulate single cells combined with parallel microbial cultivation under low nutrient flux conditions. Under these conditions, single encapsulated cells grow and form microcolonies within the microcapsules. Flow cytometry is used as a sensitive tool to detect growth within the microcapsules. Microcapsules that contain microcolonies (originated from a single encapsulated cell) are sorted individually into microtiter dishes containing organic-rich medium. This high-throughput cultivation can provide more than 10,000 bacterial and fungal isolates per environmental sample. PMID- 16260289 TI - Chemotaxis and behavioral physiology of not-yet-cultivated microbes. AB - Chemotaxis assays provide a rapid and efficient means of (1) studying the chemotactic behavior of microorganisms in complex samples and (2) identifying potential growth substrates and generating inocula for subsequent isolation trials. The chemotaxis method thus complements the set of techniques currently available for the investigation of not-yet-cultured microbes. Although restricted to motile and chemotactically active microorganisms, a considerable fraction of species can be covered with this technique, particularly in bacterioplankton communities. Several formats of the chemotaxis assay have been developed. Capillaries are loaded with solutions of test compounds and are inserted in small microscopic chambers, in bottles containing culture suspensions, or incubated directly in situ. The latter two techniques are also suitable for experiments with anaerobic bacteria. In flat rectangular glass capillaries, the accumulating microorganisms can be observed directly by light microscopy in a dark field. Afterward, the chemotactically active bacteria can be identified by analyses of their 16S rRNA gene fragments. The method has been used to identify an essential carbon compound required for the growth of previously unculturable phototrophic consortia. This knowledge proved essential for the subsequent successful enrichment of these bacterial associations. Furthermore, it has been shown that different not-yet-cultured members of aerobic lake water bacterioplankton communities are chemotactically active and attracted by different carbon compounds. PMID- 16260290 TI - Analysis of microbial communities with electrochemical microsensors and microscale biosensors. AB - Electrochemical microsensors for O2, pH, H2S, H2, and N2O are now available commercially, thus it has become a relatively simple task to analyze the microenvironment in stratified microbial communities for several chemical species. In addition, sensors are available for the physical parameters diffusivity and flow, and based on knowledge about both transport processes and microdistribution of chemistry, it becomes possible to calculate the spatial distribution and local rates of transformations, such as aerobic respiration or denitrification. As compared to other advanced techniques, microsensor equipment is inexpensive. For example, it is possible to start working with oxygen microsensors with an investment of only about 5000 dollars. Construction of one's own microsensors is only recommended for the very dedicated user, but the investment here is mainly in terms of man-hours as the equipment is simple and inexpensive. By establishing a microsensor construction facility, it is possible to work with short-lived sensors such as ion-selective microsensors for H+, NO2( ), NO3(-), Ca2+, and CO3(-) based on ion exchangers and with microscale biosensors for NO(x)-, NO2(-), CH4, and volatile fatty acids based on immobilized bacteria. PMID- 16260291 TI - Optical microsensors for analysis of microbial communities. AB - Fiber-optic microprobes connected to sensitive light meters are ideal tools to resolve the steep gradients of light intensity and spectral composition that prevail in aggregates and surface-associated microbial communities in sediments, biofilms, and microbial mats. They allow for a detailed mapping of light fields and enable insights to the complex optical properties of such highly light scattering and -absorbing microbial systems. Used in combination with microsensors for chemical species, fiber-optic irradiance microprobes allow for detailed studies of photosynthesis regulation and of the photobiology of microbial phototrophs in intact samples under ambient microenvironmental conditions of the natural habitat. Fiber-optic microprobes connected to sensitive fluorometers enable microscale fluorescence measurements, which can be used to map (i) diffusivity and flow; (ii) distribution of photosynthetic microbes, via their photopigment autofluorescence; and (iii) activity of oxygenic photosynthesis via variable chlorophyll fluorescence measurements. Furthermore, by immobilizing optical indicator dyes on the end of optical fibers, fiber-optic microsensors for temperature, salinity, and chemical species such as oxygen, pH, and CO2 can be realized. PMID- 16260292 TI - In situ measurements of metabolite fluxes: microinjection of radiotracers into insect guts and other small compartments. AB - In microbial ecology, it is of great interest to determine metabolic activities under in situ conditions, i.e., without disturbing the structure of the community and the spatial arrangement of individual populations by experimental manipulation. Microinjection of radiotracers and subsequent analysis using the isotope dilution technique has proven to be a powerful method to measure metabolic fluxes in small biological systems, e.g., the intestinal tract of termites. The large variety of commercially available radiolabeled substrates and the identification and quantitation of radiolabeled products by chromatographic methods allow for investigation of the complete metabolic network in a given system. PMID- 16260293 TI - Assaying for the 3-hydroxypropionate cycle of carbon fixation. AB - The 3-hydroxypropionate cycle is a novel pathway for autotrophic CO2 fixation, which has been demonstrated in the thermophilic phototrophic bacterium Chloroflexus aurantiacus; a yet to be defined variant of this pathway occurs in autotrophic members of the Sulfolobales (Crenarchaeota). The 3-hydroxypropionate cycle consists of the conversion of acetyl-CoA to succinyl-CoA, via malonyl-CoA, 3-hydroxypropionate, propionyl-CoA, and methylmalonyl-CoA. Carboxylation of acetyl-CoA and propionyl-CoA by acetyl-CoA/propionyl-CoA carboxylase are the CO2 fixation reactions. Succinyl-CoA serves as a precursor of cell carbon and also as a precursor of the starting compound acetyl-CoA. In C. aurantiacus, the cycle is completed by converting succinyl-CoA to malyl-CoA and cleaving malyl-CoA to acetyl-CoA and glyoxylate. Glyoxylate is then converted in a second cyclic pathway to pyruvate, which serves as a universal cell carbon precursor. The fate of succinyl-CoA in Sulfolobales is at issue. Assays used to study the characteristic enzymes of this novel pathway in C. aurantiacus are reported. PMID- 16260294 TI - Analysis of trace hydrogen metabolism. AB - H2 maintains an important role in bacterial metabolism in anaerobic environments. The ability to measure and analyze H2 concentrations in biological systems is often useful for determining the physiological state of the microbiota. Methods for precisely analyzing H2 in bacterial cultures and environmental samples are now available. This chapter discusses H2 measurements from both the theoretical and practical methodology perspective of the analysis and the interpretation of data. PMID- 16260295 TI - Advances in microscopy: microautoradiography of single cells. AB - Microautoradiography (MAR) is an efficient method to obtain reliable information about the ecophysiology of microorganisms at the single cell level in mixed communities. Data obtained by the traditional MAR method can now be improved significantly when MAR is combined with fluorescence in situ hybridization (FISH) with oligonucleotide probes for the identification of target organisms. This chapter discusses how to use MAR-FISH in various ecosystems with emphasis on the type of information to be obtained, incubation conditions, and detailed protocols for the MAR technique. It also discusses new MAR applications and the type of information that can be obtained, e.g., the use of dual labeling to investigate the simultaneous uptake of several substrates by individual prokaryotes. PMID- 16260296 TI - Atomic force microscopy of bacterial communities. AB - This chapter discusses atomic force microscopy (AFM) for the benefit of microbiologists who are interested in using this technique to examine the structures and dynamics of bacteria. AFM is a powerful technique for imaging biological samples at the nanometer to micrometer scale under nondestructive conditions. In order to be imaged with AFM, bacteria must be supported by a surface, which presents challenges because many laboratory strains of bacteria are planktonic. Still, in nature many bacteria live at surfaces and interfaces. This chapter discusses the benefits and difficulties of different methods that have been used to support bacteria on surfaces for AFM imaging and presents two methods in detail used to successfully grow and image bacteria at solid-liquid and solid-air interfaces. Using these methods it is possible to study bacterial morphology and interactions in a native state. These explorations by AFM have important applications to the study of different kinds of bacteria, interfacial bacterial communities, and biofilms. PMID- 16260297 TI - Nucleic acid recovery from complex environmental samples. AB - Effective extraction of nucleic acid from environmental samples is an essential starting point in the molecular analysis of microbial communities in the environment. However, there are many different extraction methods in the literature and deciding which one is best suited to a particular sample is very difficult. This article details the important steps and choices in deciding how to extract nucleic acids from environmental samples and gives specific details of one method that has proven very successful at extracting DNA and RNA from a range of different samples. PMID- 16260298 TI - The application of rarefaction techniques to molecular inventories of microbial diversity. AB - With the growing capacity to inventory microbial community diversity, the need for statistical methods to compare community inventories is also growing. Several approaches have been proposed for comparing the diversity of microbial communities: some adapted from traditional ecology and others designed specifically for molecular inventories of microbes. Rarefaction is one statistical method that is commonly applied in microbial studies, and this chapter discusses the procedure and its advantages and disadvantages. Rarefaction compares observed taxon richness at a standardized sampling effort using confidence intervals. Special emphasis is placed here on the need for precise, rather than unbiased, estimation methods in microbial ecology, but precision can be judged only with a very large sample or with multiple samples drawn from a single community. With low sample sizes, rarefaction curves also have the potential to lead to incorrect rankings of relative species richness, but this chapter discusses a new method with the potential to address this problem. Finally, this chapter shows how rarefaction can be applied to the comparison of the taxonomic similarity of microbial communities. PMID- 16260299 TI - Culture-independent microbial community analysis with terminal restriction fragment length polymorphism. AB - Terminal restriction fragment length polymorphism is a polymerase chain reaction (PCR)-based technique that has been used to effectively interrogate microbial communities to determine the diversity of both phylogenetic and functional markers. It requires the isolation of community DNA and knowledge of the target sequence. PCR amplification, performed with fluorescently labeled primers, is followed with restriction digestion and size selection on automated sequencing systems. The fluorescent tag identifies the terminal fragment, and the length polymorphism of the terminal fragments reveals a fraction of the phylogenetic diversity within the target sequence. Because the technique has high-throughput capabilities, it performs well in surveys where a large number of samples must be interrogated to ascertain spatial or temporal changes in community structure. PMID- 16260300 TI - Quantitative community analysis: capillary electrophoresis techniques. AB - This chapter presents methodologies for RNA extraction from soils coupled with competitive reverse transcription-polymerase chain reaction and capillary electrophoresis techniques. Combined, these approaches provide new capabilities to quantify gene expression in different environments and can aid our understanding of not only community composition, but also community activity. Such information will prove important for enhancing our knowledge of how microbial communities respond to changing geochemical parameters (e.g., temperature, pH, redox conditions, substrate levels) in situ. PMID- 16260301 TI - In situ functional gene analysis: recognition of individual genes by fluorescence in situ hybridization. AB - Fluorescence in situ hybridization (FISH) using specific probes certainly is one of the most commonly applied molecular techniques in microbial ecology. Monitoring of community composition and dynamics can be combined with localization and identification of individual cells in situ. However, the resolution power of the method is limited by the need for high target numbers per cell. Apart from standard targets (ribosomal RNAs), mRNAs could be used successfully for in situ visualization in some cases. A new promising variant of in situ hybridization could be established that should provide access to any low copy number nucleic acid targets, such as chromosomal genes. The recognition of individual genes by FISH (RING-FISH) technology is based on target visualization mediated by polynucleotide probe network formation. The specificity of the approach is provided by intracellular probe-target hybridization. This initial hybridization apparently acts as a focal point for inter-probe hybridization within and mainly in the periphery of the cells. Probe-conferred fluorescence typically appears halo-like around the cells. RING-FISH can be used in combination with conventional oligonucleotide FISH. Thus, genetic potential can be assigned to in situ identified cells. PMID- 16260302 TI - Simultaneous fluorescence in situ hybridization of mRNA and rRNA for the detection of gene expression in environmental microbes. AB - A protocol is presented for the detection of gene expression in environmental microorganisms by means of fluorescence in situ hybridization (FISH). Messenger RNA (mRNA) is hybridized with digoxigenin (DIG)- or fluorescein (FLUOS)-labeled ribonucleotide probes. Subsequently the hybrid is detected immunochemically with a horseradish peroxidase (HRP)-labeled antibody and tyramide signal amplification (catalyzed reporter deposition, CARD). After mRNA FISH, microorganisms can be identified by rRNA FISH with oligonucleotide probes labeled either with a fluorochrome or with HRP. Sample preparation and cell permeabilization strategies for various microbial cell types are discussed. The synthesis of DIG- and FLUOS labeled probes, as well as custom labeling of tyramides with different fluorochromes, is described. As a case study, we describe in detail mRNA FISH of the particulate methane-monooxygenase, subunit A (pmoA) in endosymbiotic bacteria from tissue sections of a marine mollusc. PmoA is used as a marker gene for methanotrophy. PMID- 16260303 TI - Community-level analysis of phototrophy: psbA Gene Diversity. AB - Photosynthetic organisms play a crucial role in the marine environment. In vast areas of the oceans, most of this marine production is performed by cells smaller than 2-3 microm (picoplankton). This chapter describes molecular analyses of the conserved photosynthetic psbA gene (protein D1 of photosystem II reaction center) as a diversity indicator of naturally occurring marine oxygenic picophytoplankton and of marine cyanophages carrying photosynthesis genes. PMID- 16260304 TI - Quantitative analysis of nifH genes and transcripts from aquatic environments. AB - The availability of fixed inorganic nitrogen often plays a fundamental role in regulating primary production in both aquatic and terrestrial ecosystems. Because biological nitrogen fixation is an important source of nitrogen in marine environments, the study of N2-fixing microorganisms is of fundamental importance to our understanding of global nitrogen and carbon cycles. Quantitative molecular tools have made it possible to examine uncultivated N2-fixing microorganisms directly in the environment. Currently, we are using quantitative polymerase chain reaction (PCR; Q-PCR) and quantitative reverse transcriptase PCR (Q-RT-PCR) to study the ecology and gene expression of N2-fixing bacteria in aquatic environments. Using these methods, we discovered that specific estuarine diazotrophs have distinct nonrandom distributions and that some diazotrophs in the open ocean have different diel patterns of nifH gene expression. This chapter describes briefly our 5' nuclease assay protocols for Q-PCR and Q-RT-PCR of nifH gene fragments in environmental samples and discusses some important methodological considerations for the quantitative molecular examination of microbes in aquatic environments. PMID- 16260305 TI - Community level analysis: genetic and biogeochemical approaches to investigate community composition and function in aerobic ammonia oxidation. AB - Aerobic ammonia oxidation is the process that converts ammonium to nitrate and thus links the regeneration of organic nitrogen to fixed nitrogen loss by denitrification. It is performed by a phylogenetically restricted group of Proteobacteria (ammonia-oxidizing bacteria, AOB) that are autotrophic and obligately aerobic. This chapter describes methods for the measurement of ammonia oxidation in the environment, with a focus on seawater systems and stable isotopic tracer methods. It also summarizes the current state of molecular ecological approaches for detection of AOB in the environment and characterization of the composition of AOB assemblages. PMID- 16260306 TI - Community-level analysis: key genes of aerobic methane oxidation. AB - Aerobic methane-oxidizing bacteria (methanotrophs) are a diverse group of bacteria that are currently represented by 13 recognized genera. They play a major role in the global methane cycle and are widespread in nature with representatives found in soils, freshwater, seawater, freshwater and marine sediments, peat bogs and at extremes of temperature, salinity, and pH. There has been an interest in methanotrophs for their potential in bioremediation processes. Methanotroph diversity and ecology are often studied using the "functional" genes pmoA, mmoX, and mxaF, encoding subunits of the particulate methane monooxygenase, soluble methane monooxygenase, and the methanol dehydrogenase, respectively. This chapter describes methods used to detect and analyze these functional genes. PMID- 16260307 TI - Methyl-coenzyme M reductase genes: unique functional markers for methanogenic and anaerobic methane-oxidizing Archaea. AB - In many anoxic environments, methanogenesis is the predominant terminal electron accepting process involved in the mineralization of organic matter, which is catalyzed by methanogenic Archaea. These organisms represent a unique but phylogenetically diverse guild of prokaryotes, which can be conveniently tracked in the environment by targeting the mcrA gene as a functional marker. This gene encodes the alpha subunit of the methyl-coenzyme M reductase (MCR), which catalyzes the last step in methanogenesis and is present in all methanogens. Cultivation-independent analysis of methanogenic communities involves the polymerase chain reaction (PCR) amplification of the mcrA gene from extracted community DNA, comparative analysis of mcrA clone libraries, or PCR-based fingerprinting analysis by terminal restriction fragment polymorphism analysis (T RFLP). It has also been suggested that anaerobic methane-oxidizing Archaea possess MCR, which facilitates detection of this novel group of "reverse methanogens" as well using the mcrA gene as a functional marker. PMID- 16260308 TI - Community-level analysis: genes encoding methanopterin-dependent enzymes. AB - This chapter describes a set of novel tools for the environmental detection of C1 transfer functions linked to the cofactor methanopterin. These tools include degenerate environmental primers targeting four of the most conserved genes in the methanopterin-linked C1 transfer pathway in bacteria, fae, mtdB, mch, and fhcD, and extensive databases of the respective genes. The tools described are suitable for detecting methanopterin-linked formaldehyde-oxidizing capacity in natural microbial communities and for determining the phylogenetic affiliations of major phyla involved in single-carbon cycling in the environment. The range of detection includes a variety of methano- and methylotrophic groups, other proteobacterial species capable of methanopterin-mediated reactions, and a variety of planctomycetes, as well as groups of microbes with currently unknown phylogenetic affiliations. PMID- 16260309 TI - Community-level analysis: key genes of CO2-reductive acetogenesis. AB - CO2-reductive acetogenic bacteria are ubiquitous in anaerobic habitats and are physiologically and phylogenetically diverse. The latter characteristics have rendered their diversity in natural environments, their distributions, and their ecological function(s) difficult to assess. Recently introduced polymerase chain reaction (PCR) primers for specific amplification of the structural gene encoding formyltetrahydrofolate synthetase (FTHFS, EC 6.3.4.3), a key enzyme in the acetyl CoA pathway of acetogenesis, have facilitated studies of acetogen diversity and ecology. These primers amplify an approximately 1100-bp segment of the FTHFS gene. FTHFS sequences have been recovered from authentic acetogens, from sulfate reducing bacteria, and from a variety of other nonacetogenic bacteria. Phylogenetic analyses segregated these sequences into distinct clusters, only one of which contained sequences from known acetogens. This chapter describes the PCR primers, defines conditions for successful amplification of FTHFS sequences, and details the phylogenetic analysis of the FTHFS sequences. Information on the types of sequences that have been recovered from natural acetogen habitats and how they have been interpreted is also included. PMID- 16260312 TI - Introduction to veterinary physical rehabilitation. AB - Physical therapy is a profession with an established scientific basis in human beings and companion animals. It has a large number of clinical applications in the restoration, maintenance, and promotion of optimal physical function. In providing physical therapy, the goal is to restore, maintain, and promote optimal function, optimal fitness, wellness, and quality of life as they relate to movement disorders and health. A major emphasis is to prevent or minimize the onset, clinical signs, and progression of impairments, functional limitations, and disabilities that may result from diseases, disorders, conditions, and injuries. PMID- 16260310 TI - Functional marker genes for identification of sulfate-reducing prokaryotes. AB - Sulfate-reducing prokaryotes (SRPs) exploit sulfate as an electron acceptor for anaerobic respiration and exclusively catalyze this essential step of the world's sulfur cycle. Because SRPs are found in many prokaryotic phyla and are often closely related to non-SRPs, 16S rRNA gene-based analyses are inadequate to identify novel lineages of this guild in a cultivation-independent manner. This problem can be solved by comparative sequence analysis of environmentally retrieved gene fragments of the dissimilatory (bi)sulfite (dsrAB) and adenosine 5'-phosphosulfate reductases (apsA), which encode key enzymes of the SRP energy metabolism. This chapter provides detailed protocols for the application of these functional marker molecules for SRP diversity surveys in the environment. Data from the analysis of dsrAB sequence diversity in water samples from the Mariager Fjord in northeast Denmark are presented to illustrate the different steps of the protocols. Furthermore, this chapter describes a novel gel retardation-based technique, suitable for fingerprinting of the approximately 1.9-kb-large dsrAB polymerase chain reaction amplification products, which efficiently increases the chance of retrieving rare and novel dsrAB sequence types from environmental samples. PMID- 16260313 TI - Biomechanics of rehabilitation. AB - The biomechanics of motion and rehabilitation are complex, with many tissue types and structures involved. In addition, consideration must be given to the stage of tissue healing with some injuries, such as fractures.A more thorough knowledge of some of the infrequently discussed biomechanical aspects of musculoskeletal tissues and motion during rehabilitation, combined with known features of tissue recovery, should enhance the development of rehabilitation programs for patients. PMID- 16260314 TI - Joint mobilization. AB - Therapeutic touch has been used in human beings to soothe aches and pains. Most dogs also seem to enjoy being touched. Manual therapy techniques are skilled hand movements intended to improve tissue extensibility; increase range of motion; induce relaxation; mobilize or manipulate soft tissue and joints; modulate pain; and reduce soft tissue swelling, inflammation, or restriction. The intent of this article is to provide an overview of the principles of manual therapy, followed by selected treatment techniques for the hip, stifle, elbow, shoulder, carpus.and thoracic and lumbar spine. The techniques of G.D. Maitland, an Australian physical therapist who developed a clinically based approach in the 1960s and 1970s, are emphasized. PMID- 16260315 TI - Physical agent modalities. AB - The purpose of this article is to review the use of cold, heat, therapeutic ultrasound, and electrical stimulation in small animal rehabilitation. The material in this article is a compilation from the veterinary and hu-man literature. Additional information is needed on how to adapt the techniques used in human beings to small animals and then to establish the efficacy of these techniques in animals. PMID- 16260316 TI - Emerging modalities in veterinary rehabilitation. AB - Many new modalities have been introduced in human and veterinary physical rehabilitation. In many instances, there is sound theory of how they may impact the physiology of various cells, tissues, or organs. This article reviews some of the modalities that have been introduced recently in human and veterinary rehabilitation. Topics include low-level laser, phototherapy, and extracorporeal shock wave treatment. PMID- 16260317 TI - Rehabilitation for the orthopedic patient. AB - An understanding of orthopedic conditions and their medical and surgical treatment is important to help the therapist develop a treatment plan that will help the patient return to function quickly with minimal complications. The therapist must constantly assess the patient for improvement or complications and adjust the therapy plan accordingly. Knowledge of the stages of tissue healing and of the strength of tissues is critical to avoid placing too much stress on the surgical site, yet some challenge to tissues must be provided to optimize the return to function. PMID- 16260318 TI - Rehabilitation for the neurologic patient. AB - A properly designed rehabilitation program should be an important component of the treatment plan of animals with neurologic disease. Such a program should be designed in conjunction with appropriate treatment of the underlying problem and after special consideration of the origin of the neurologic problem, the severity of the signs, the cause of the signs, their anticipated progression, and the needs of the owner and the pet. This article describes the pathophysiology of injury and recovery in the central and peripheral nervous systems, assessment of the neurologic patient, data on the prognosis and expected course of recovery for a variety of different diseases, and rehabilitation exercises appropriate for neurologic patients. PMID- 16260319 TI - Rehabilitation of medical and acute care patients. AB - Cancer and serious systemic illness result in several physiologic changes that involve multiple body systems. While the primary conditions are addressed with traditional modalities of medicine, the side effects, secondary changes, and complications can be ameliorated or even prevented with rehabilitation and supportive care. This article reviews problems facing the oncologic and critically ill animal, discusses basic techniques in the management of these animals, and highlights the essential role of rehabilitation in obtaining maximal functional capacity in the critically ill patient. PMID- 16260320 TI - Rehabilitation and conditioning of sporting dogs. AB - Owners and trainers exercise sporting dogs to increase their fitness and optimize their conditioning and performance. Training is designed to in-crease strength, endurance, and agility and is sport-specific. Sporting dogs are susceptible to specific musculoskeletal injuries. The rehabilitation of sporting dogs after these injuries follows specific principles during the acute, subacute, and reconditioning periods. PMID- 16260321 TI - Assistive devices, orthotics, and prosthetics. AB - Deciding on which supportive device, orthotic, or prosthetic is best suited for a given patient is a complex process involving many different factors. The ability to manage biomechanical abnormalities successfully may be enhanced by an understanding of the properties of the various materials that comprise these devices, their effect on functional performance, and other associated patient factors. Veterinary health care providers are faced with the challenge of effectively addressing the physiologic and fiscal needs of the patient in a rapidly changing patient care environment. PMID- 16260322 TI - Wound healing in the veterinary rehabilitation patient. AB - Wound healing is a biologically complex cascade of predictable overlap-ping events and is a natural restorative response to tissue injury. The biologic process for wound healing is the same for all wounds, although the specific mechanisms may vary. This article reviews the wound heal-ing process, discussing factors that may delay normal healing progression and potential modalities and treatments to aid healing. PMID- 16260323 TI - Logistics of companion animal rehabilitation. AB - Setting up rehabilitation services presents several challenges to the veterinary practitioner. Members of the rehabilitation team need to have a solid knowledge base of acute and chronic orthopedic and neurologic disorders and a knowledge of rehabilitation principles and applications. The working environment has to be adapted to patients with limited mobility, specific equipment and supplies are to be used, and a fee structure should be implemented. This article reviews these logistic aspects of companion animal rehabilitation. PMID- 16260324 TI - The practice of infection control and applied epidemiology: a historical perspective. AB - The United States health care system and patient populations have changed substantially over the past several decades. The practice of infection control also has evolved since the landmark Study on the Efficacy of Nosocomial Infection Control project, and infection control professionals (ICPs) must continue to develop the knowledge and skills necessary to practice infection prevention and control. Practice analyses of infection control conducted between 1982 and 2001 were analyzed to determine changes in practice. These data reflect a 145% increase in infection control activities over a 20-year period. However, resources for infection control and prevention have not kept pace with this increased activity. In addition, the current trend toward mandatory reporting of health care-associated infections (HAIs) among several states will add more tasks for ICPs with limited resources, at the risk of spending less time on prevention and control activities. In keeping with its philosophy of quality health care and responsible public reporting, the Association of Professionals in Infection Control and Epidemiology, Inc, continues to explore the issue of mandatory reporting of HAIs. PMID- 16260325 TI - Systematic review of economic analyses of health care-associated infections. AB - BACKGROUND: Economic evidence is needed to assess the burden of health care associated infections (HAIs) and cost-effectiveness of interventions aimed at reducing related morbidity and mortality. The objective of this study was conducted to assess the quality of economic evaluations related to HAI and synthesize the evidence. METHODS: A systematic review of research published between January 2001 and June 2004 was conducted. Quality of the publication was estimated using a Likert-type scale. All cost estimates were standardized into a common currency. Descriptive statistics and a logistic regression were conducted to identify predictors of high quality. RESULTS: 70 studies were audited. There was wide variation in these cost estimates. Publications estimating the cost attributable to an infection were almost 7 times more likely judged to be of higher quality than studies of the cost of interventions (P < .05). Papers in which the authors stated the perspective (hospital or societal) were twice as likely to be judged as being of high quality (P < .05). CONCLUSION: There are more publications and growing interest in estimating the costs of HAI. However, the methods employed vary. We recommend (1) the use of guidelines for authors and editors on conducting an economic analysis, (2) development of more sophisticated mathematical models, and (3) training of infection control professionals in economic methods. PMID- 16260326 TI - Hepatitis B vaccination in correctional health care workers. AB - BACKGROUND: Data on bloodborne pathogen risk among health care workers (HCWs) employed in the correctional setting are sparse, even though the prevalence of bloodborne infections, including hepatitis B virus (HBV), among inmates is high. To address this, we determined prevalence and correlates of hepatitis B virus vaccination status in correctional health care workers (CHCWs) employed in 3 state correctional health care facilities. METHODS: A confidential, self administered cross-sectional survey was performed. RESULTS: Four hundred eleven (69.8%) of 588 eligible participants completed the survey. Of these, 264 (64.2%) reported receiving a primary hepatitis B (HB) vaccine series. Vaccination rates varied by state and by job category. Parenteral exposures were not uncommon; 8.6% (n = 24) of clinical CHCWs and 2.0% (n = 7) of nonclinical CHCWs reported one or more needlesticks in the 6-month period prior to the study. Among clinical staff, vaccination correlated with licensure (RN or MD) and race (white) and in nonclinical staff with history of close contact with HBV infected inmates and with needlestick injury. CONCLUSION: Although the HB vaccination rate among CHCWs was generally high, given their potential risk of exposure to HBV, universal vaccination should be encouraged and should include those nonclinicians with job duties that may involve potential exposure to blood/body fluids. PMID- 16260328 TI - A review of the possible role of oral and dental colonization on the occurrence of health care-associated pneumonia: underappreciated risk and a call for interventions. PMID- 16260327 TI - Tuberculosis in health care workers during declining tuberculosis incidence in New York State. AB - BACKGROUND: Nosocomial tuberculosis (TB) transmission has decreased dramatically in New York State since 1992; however, health care workers (HCWs) still compose >3% of TB cases. METHODS: Aggregate surveillance data on incident TB cases from 1994 to 2002 were examined for trends among HCWs. Additional information was available for HCW cases from 1998 to 2002, including facility type, tuberculin skin test (TST) result at hire, and treatment of latent TB infection (TLTBI). RESULTS: In New York State, 2.5% of TB cases in 1994 and 4.0% in 2002 were in HCWs (P value for trend <.001). Fifty percent of HCWs TB cases in 1994 and 77.6% in 2002 were in non-US born (P = .002) HCWs. Multidrug-resistant TB in HCWs decreased from 15.6% in 1994 to 6.9% in 2002 (P = .001). Of 297 HCWs TB cases in 1998-2002, 54.9% were TST positive at hire, and 21.2% had unknown TST result; 50.2% of 221 HCWs who were TST positive at or after hire met guidelines for TLTBI, and 23.4% received treatment. The highest proportion with unknown TST at hire and the lowest proportion receiving TLTBI were in ambulatory facilities. CONCLUSION: Many HCWs who developed TB were either TST positive at hire and did not receive TLTBI or did not receive TST at hire. Facilities should encourage treatment for HCWs who meet criteria for TLTBI. Provider education should focus on ambulatory facilities. PMID- 16260329 TI - The economic impact of infection control: making the business case for increased infection control resources. PMID- 16260330 TI - State-of-the-science--2004: time for a "No Excuses/No Tolerance" (NET) strategy. PMID- 16260332 TI - Bacterial contamination of original microbiology culture reports in a community hospital. PMID- 16260331 TI - Comparative efficacy of hand hygiene agents in the reduction of bacteria and viruses. PMID- 16260333 TI - A pilot study on the transfer of 137Cs and 90Sr to horse milk and meat. AB - The radiological assessment of the impact of nuclear weapon's testing on the Semipalatinsk Test Site (STS) on the local population requires comprehensive site specific information on radionuclide behaviour in the environment. However, information on radionuclide behaviour in the conditions of the STS is rather sparse and, in particular, there are no data in the literature on parameters of radionuclide transfer from feed to horse products proofed to be important contributors to the internal dose to the local population. The transfer of 137Cs and 90Sr to horse milk and meat was studied under laboratory and field conditions: in controlled experiment with three lactating horses maintained in the Kazakh Agricultural Research Institute, and in field measurements of horse products taken from horses grazing at the Semipalatinsk Test Site. The equilibrium transfer factors from feed to horse milk and meat were estimated to be 0.012 dl(-1) and 0.035 dkg(-1) for (137)Cs and 0.0022 dl(-1) and 0.003 dkg(-1) for (90)Sr, respectively. The biological half-lives were approximated by a sum of two exponentials amounting to 3 (85%) and 15 (15%) days for 137Cs and 3.5 (70%) and 100 (30%) days for 90Sr. The highest 137Cs transfer has been found to be to spleen, followed by lung, heart, muscles, kidneys, intestine, and finally skin and bones. For90Sr, the maximum activity concentration was observed in bones; contamination of other tissues is rather uniform except for liver and intestine with a factor of about 2 higher than muscles. PMID- 16260335 TI - Neonatal and maternal outcomes in twin gestations > or =32 weeks according to the planned mode of delivery. AB - OBJECTIVE: To determine a possible relationship between neonatal and maternal outcomes in twin gestations and the planned mode of delivery. STUDY DESIGN: A single-centre retrospective cohort study in twins > or =32 weeks of gestational age was performed. Baseline characteristics, and neonatal and maternal outcomes were documented according to the planned mode of delivery: a planned caesarean section or a planned vaginal birth. Statistical analysis was performed using chi square test. Fisher exact test was used in case correction was needed. RESULTS: During the study period (1999-2002), 164 twins > or =32 weeks were enrolled in the study. In 29 women (17.7%) an elective caesarean section was performed. The remaining 135 twins (82.3%) were allowed to start a vaginal delivery. An emergency or an urgent secondary caesarean section for both twins was performed in 26 women, and in 2 women for twin B only. One twin B baby died during planned vaginal delivery. No significant differences in perinatal mortality and serious neonatal morbidity were found between both groups (10.3% versus 9.6%). Neonatal outcomes in twins A were significantly better than in twins B (2.4% versus 7.3%), independent of the planned mode of delivery. Serious maternal morbidity was not significantly different between both groups (13.8% versus 19.3%), although 2 women in the elective caesarean section group needed a relaparotomy for haemorrhage. CONCLUSION: Our results do not support an elective caesarean section for twin gestations > or =32 weeks. The success rate of vaginal delivery in the planned vaginal birth group was nearly 80%. PMID- 16260336 TI - Is epidural analgesia a risk factor for occiput posterior or transverse positions during labour? AB - OBJECTIVE: The aim of this study was to assess whether the station of the fetal head at epidural placement is associated with the risk of malposition during labour. STUDY DESIGN: Retrospective study (covering a 3-month period) of patients in labour with singleton cephalic term fetuses and epidural placement before 5 cm of dilatation. We studied the following risk factors for malposition: station and cervical dilatation at epidural placement, induction of labour, parity and macrosomia. Malposition, defined as all occiput posterior and occiput transverse positions, was assessed at 5 cm of dilatation because of our policy of systematic manual rotation for malpositions. RESULTS: The study included 398 patients, 200 of whom had malpositions diagnosed at 5 cm of dilatation. In both the univariate and multivariate analyses, station at epidural placement was the only risk factor significantly associated with this malposition (adjusted OR: 2.49, 95% CI 1.47 4.24). None of the other factors studied was significantly associated with malposition: nulliparity (OR 1.45, 95% CI 0.96-2.20), macrosomia (OR 0.75, 95% CI 0.37-1.50), induction of labour (OR 0.84, 95% CI 0.49-1.45), or dilatation less than 3 cm at epidural administration (OR 1.16, 95% CI 0.59-2.30). Only three infants of the 365 delivered vaginally (0.8%) were born in occiput posterior positions. CONCLUSION: Epidural placement when the fetal head is still "high" is associated with an increased rate of occiput posterior and transverse malpositions during labour. PMID- 16260337 TI - Haemoglobin and serum ferritin in pregnancy--correlation with smoking and body mass index. AB - BACKGROUND: Little is known about factors which may influence haemoglobin (Hb) and ferritin levels in pregnancy. AIM: To analyse if haemoglobin and ferritin levels during pregnancy are influenced by maternal age, body mass index, cigarette smoking, and iron supplementation. METHODS: A random sample of 561 parous pregnant women were recruited from the catchment areas of three Scandinavian university hospitals. The analyses were based on 5024 haemoglobin and 1529 ferritin measures sampled from the first trimester to 42 weeks of gestation. Multilevel modelling was used to construct mean and percentile curves for haemoglobin and ferritin by gestational age. RESULTS: Women aged 25-34 years had significantly higher haemoglobin values than older and younger women. Haemoglobin values were significantly lower for women with body mass index < 19 kg/m(2) than for women with body mass index > or =19. Smokers had significantly lower haemoglobin values throughout pregnancy compared to non-smokers, with the lowest values among women who smoked 1-9 cigarettes per day. There were no similar associations between ferritin and maternal age, body mass index, or smoking. Women with iron supplementation throughout pregnancy had a higher relative increase in haemoglobin concentration toward the end of pregnancy. In non-supplemented women the decline in ferritin concentration was significantly steeper than in those who received iron supplementation. CONCLUSIONS: Haemoglobin levels during pregnancy are significantly associated with maternal age, cigarette smoking, body mass index, and iron supplementation. No such associations were found with ferritin levels, except for iron supplementation. PMID- 16260339 TI - Diagnostic criteria and reporting procedures for pre-eclampsia: a national survey among obstetrical departments in Denmark. AB - OBJECTIVE: A precondition for the rational use of obstetric databases in biomedical research is detailed knowledge on how data are being generated. We identified the diagnostic procedures and criteria for pre-eclampsia (PE) and assessed the level of obstetric training of the personnel responsible for the records submitted to the patient registry at the Danish National Board of Health. STUDY DESIGN: A structured questionnaire, including three case stories, was sent to the chief consultant of the department. RESULTS: Thirty-three out of the 34 Danish departments (97%) returned the questionnaire. Reporters of pregnancy diagnoses to the National Patient Registry differed widely in training. For complicated pregnancies, departments ranged from having only specialists reporting all cases to secretaries reporting up to 50%. Cut off limits of blood pressure (BP) and protein loss used to diagnose pre-eclampsia showed large differences across departments. The diagnoses given to three case stories showed little correlation to the criteria the departments reported using. CONCLUSION: Even in a small country like Denmark with 34 obstetrical departments, there was little consensus on the diagnostic criteria for pre-eclampsia. The findings emphasize the need for standardizing diagnostic criteria and reporting practice and may have implications on how to interpret data regarding pre-eclampsia. PMID- 16260338 TI - Vascular expression of adrenomedullin is increased in Wistar rats during early pregnancy. AB - OBJECTIVE: Circulating levels of adrenomedullin (ADM)--a vasodilator peptide with long-lasting effects--increase in the course of pregnancy. Neither the site nor the concomitant rate of ADM synthesis in pregnancy is known. The aim of this study was to test the hypothesis that the rise in plasma levels of ADM during pregnancy is paralleled by increased gene expression and protein levels in the vascular bed. STUDY DESIGN: We determined in cardiovascular and reproductive tissues of non-pregnant (n=10) and 10-days pregnant (n=10) Wistar rats ADM gene expression by semi-quantitative RT-PCR (normalized to GAPDH). As a support for the mRNA data, protein concentrations were measured by both ELISA and Western blot analysis. Finally, ADM in these tissues was localized by immunohistochemical staining. Statistical analysis was carried out by applying Mann-Whitney U-test. RESULTS: ADM mRNA levels in the abdominal aorta, renal artery and the kidney were increased during pregnancy. In addition, immunohistochemical staining in the kidney, uterus, abdominal aorta, renal, uterine and superior mesenteric artery was more intense as compared to non-pregnant rats. However, we observed lower concentrations of tissue ADM protein in pregnant rats, indicating an increased release of the hormone by the producing cells. CONCLUSION: Vascular ADM gene expression is increased in the first half of rat pregnancy. This coincides and may be functionally related to the institution of a high flow/low resistance circulation in pregnancy. PMID- 16260340 TI - Fluctuations in C-reactive protein concentration and neutrophil activation during normal human pregnancy. AB - OBJECTIVES: To clarify the changes in serum C-reactive protein (CRP) levels and in the neutrophil activation state during normal human pregnancy. MATERIALS AND METHODS: A longitudinal study (n=23) was performed during the three trimesters of pregnancy; a group of non-pregnant women (n=24) was used as control. Total and differential leukocyte count, serum concentration of CRP and plasma levels of granulocyte-macrophage colony stimulating factor (GM-CSF) and of lactoferrin and elastase (two indirect markers of neutrophil activation) were measured. RESULTS: Pregnancy imposed an inflammatory response in the mother, observed by the significant increment in total white blood cell (WBC) and neutrophil counts and in the circulating levels of CRP, GM-CSF and lactoferrin, in all trimesters of gestation compared with non-pregnant controls. Plasma elastase concentration was also significantly higher in pregnant women, but only in the first trimester of gestation. Regarding the ratios of lactoferrin and elastase per neutrophil, they were significantly lower in pregnant women (all trimesters). During gestation, WBC and neutrophil count increased significantly from the first to the second trimester and remained high in the third period. In contrast, the ratios of lactoferrin and elastase per neutrophil decreased significantly from the first to the second trimester, remaining low in the last trimester. Concerning CRP levels, no consistent changes were observed throughout gestation; 12 cases (52.2%) presented fluctuations, whereas 7 (30.4%) showed progressive reductions and 4 (17.4%) progressive increments throughout pregnancy. CONCLUSIONS: Changes in CRP levels vary in a wide manner between subjects along pregnancy, even though median values are consistently elevated throughout pregnancy. Moreover, circulating levels of neutrophil-activation products are higher in normal human gestation. PMID- 16260341 TI - Factors affecting survival in infants weighing 750 g or less. AB - OBJECTIVE: To assess which factors independently affect survival in infants weighing 750g or less. STUDY DESIGN: We reviewed the obstetric, neonatal, and placental pathology information of all non-malformed neonates with birth weight of 750g or less from January 1998 to December 2002. Logistic regression analysis was used to control for the effect of confounding variables. A P<0.05 was considered significant. RESULTS: Fifty nine neonates fulfilled the inclusion criteria; 30 (51%) survived the perinatal period. Surviving neonates were more frequently born after steroid administration (P=0.03) and from indicated delivery (P=0.01), had greater birth weight (P=0.001), gestational age at delivery (P<0.001), and 5-min Apgar scores of 7 or more (P=0.04) than those who died. There were no significant differences in placental pathology between survivors and neonates who died. Stepwise logistic regression analysis showed that gestational age (P=0.01), birth weight (P=0.004), female sex (P=0.03), 5-min Apgar score (0.026), and steroid administration (P=0.04) were independent predictors of survival. Cumulatively these five predictors explained 69% of neonatal survival. CONCLUSIONS: The predictors of survival among micropremies are the same as those reported for older preterm neonates. The type of preterm delivery (spontaneous versus indicated) and placental pathology do not independently affect survival. PMID- 16260342 TI - Preference and acceptability of oral versus vaginal administration of misoprostol in medical abortion with mifepristone. AB - OBJECTIVES: To compare the experience of pain, need of analgesic interventions, preference and acceptability in medical abortion up to 49 days of amenorrhea with mifepristone and orally versus vaginally administered misoprostol. STUDY DESIGN: Ninety-seven women were randomised to oral misoprostol, n=48, or vaginal misoprostol, n=49. On day 1 of the study, both the groups received 600 mg of mifepristone. On day 3 of the study, one group received 0.4 mg of misoprostol orally and the other group received 0.8 mg of misoprostol vaginally. RESULTS: Even though oral administration of misoprostol seemed to be associated with a higher rate of gastrointestinal side effects, women in both the groups showed a clear preference towards the oral route of administration. The willingness to administer the misoprostol at home was also higher among the women in the oral group, which may in part depend on a more positive/less negative experience of the abortion. CONCLUSION: A majority of women prefer oral administration of misoprostol in early medical abortion. PMID- 16260343 TI - Insulin receptor tyrosine kinase activity and substrate 1 (IRS-1) expression in human myometrium and leiomyoma. AB - BACKGROUND: Uterine leiomyomas are the commonest tumors of the genital tract. Growth factors seem to be implicated in the development of leiomyoma. OBJECTIVE: To determine the insulin receptor (IR) tyrosine kinase activity--phosphorylation of exogenous substrate poly(Glu 4: Tyr 1)--and insulin receptor substrate 1 expression in normal myometrium and leiomyoma. STUDY DESIGN: The study group consisted of 12 women with leiomyoma undergoing routine hysterectomy. Samples of leiomyoma and adjacent normal myometrium were obtained at the time of operation. Plasma membrane fractions were prepared and samples were incubated with and without insulin and incubated with exogenous substrate poly(Glu 4: Tyr 1). IRS-1 expression was studied in the whole lysate via Western blotting using specific antibodies. Data were analyzed using Student's t-test. RESULTS: The phosphorylation of the exogenous substrate poly(Glu 4: Tyr 1) in myometrium (1.566+/-0.177) and in leiomyoma (1.98+/-0.612) were similar (P=0.774). The IRS-1 levels in myometrium (0.190+/-0.022) and in leiomyoma (0.226+/-0.022) were not different (P=0.184). CONCLUSIONS: There was no difference in IR tyrosine kinase activity (phosphorylation of exogenous substrate) and IRS-1 expression between normal myometrium and leiomyomata. Other steps in the insulin signaling cascade require further study to investigate the role of insulin receptor in leiomyomata. PMID- 16260344 TI - Autoimmune progesterone dermatitis: a rare pathology. PMID- 16260345 TI - Vulvar lichen sclerosus successfully treated with topical tacrolimus. PMID- 16260346 TI - Specialist life--Janesh Gupta. PMID- 16260347 TI - Presence of the metabolic syndrome distinguishes patients with ketosis-prone diabetes who have a Type 2 diabetic phenotype. AB - OBJECTIVE: This study was designed to determine if patients with ketosis-prone diabetes could be distinguished clinically on the basis of features of the metabolic syndrome (MetS). METHODS: Participants with ketosis-prone diabetes admitted to our institution during a 3-year period were studied. One hundred seventy-two patients from four ethnic backgrounds were divided into two groups: with or without MetS, as defined by the World Health Organization. Pearson's chi square test or one-way ANOVA was used as appropriate to evaluate group differences. RESULTS: Of the 172 patients, 74 (43%) had at least three characteristics of MetS (+MetS group), while 98 (57%) had less than three characteristics of MetS (-MetS group). The +MetS group had significantly higher beta-cell functional reserve as measured by the glucagon stimulation test [area under the curve (AUC) for C-peptide levels 19.50+/-2.26 compared with 8.66+/-1.66 ng/ml over 10 min; P=.0001]. A significantly higher proportion of patients in the +MetS group had preserved beta-cell functional reserve than in the -MetS group (70% compared with 30%; P<.00001). After 12 months of follow-up, +MetS patients had significantly lower HbA1c levels (8.3+/-0.4% compared with 9.8+/-0.4%; P=.01) and higher C-peptide/glucose ratios (2.01+/-0.28 compared with 1.14+/-0.21; P=.02) than the -MetS patients did. CONCLUSIONS: Patients with ketosis-prone diabetes with the MetS are more likely to have features associated with Type 2 diabetes, despite having developed diabetic ketoacidosis (DKA), with better beta cell functional reserve and glycemic control after the index DKA episode. PMID- 16260348 TI - Are Type 2 diabetic patients offered adequate foot care? The role of physician and patient characteristics. AB - In the context of a nationwide outcomes research program on Type 2 diabetes, we investigated physician and patient practices related to foot care. Patients filled in a questionnaire investigating whether they had received information about foot care, how often they had had their feet examined in the last year, and how often they usually checked their feet. Analyses were adjusted for patient case-mix and physician-level clustering. Overall, 3564 patients were recruited by 125 diabetes outpatient clinics (DOCs) and 103 general practitioners (GPs), of whom 6.8% suffered form lower limb complications. The presence of foot complications was correlated with insulin treatment, cigarette smoking, low levels of school education, and the presence of other diabetic complications. More than 50% of the patients reported that they had not had their feet examined by their physician and 28% referred that they had not received foot education. Patients with lower levels of school education and income, as well as overweight individuals, were less likely to receive foot education. Physicians tended to perform foot examination more often in males, low-income patients, those with foot complications, and those treated with insulin, but not in patients with the highest risk of foot complications, that is, those with diabetic neuropathy or peripheral vascular disease (PVD). GPs tended to perform foot examination less frequently than diabetologists do. Foot self-examination was not performed by 33% of the patients. Those individuals who had received foot education or had had their feet examined were more likely to check their feet regularly. A substantial proportion of Type 2 diabetic patients is not offered adequate foot care, even in the presence of major risk factors for lower limb complications. Patient knowledge and practices are strongly related to physicians' attitudes. PMID- 16260349 TI - A copper chelating agent suppresses carbonyl stress in diabetic rat lenses. AB - To clarify whether transition metals are involved in carbonyl stress in diabetic tissues, we observed the effects of a metal chelating agent, trientine (TE) hydrochloride on the levels of methylglyoxal (MG), 3-deoxyglucosone (3-DG), advanced glycation end products, 8-hydroxy-2'-deoxyguanosine (8-OHdG), and polyol pathway metabolites along with semicarbazide-sensitive amine oxidase (SSAO) enzyme activity in lenses from streptozotocin-induced diabetic rats. Lens MG and 3-DG levels were significantly higher in diabetic rats than nondiabetic controls, and TE significantly restored the increase of these compounds. Lens argpyrimidine was also increased in diabetic rats as compared with controls and was significantly reduced by TE. Lens SSAO activity and 8-OHdG were also significantly elevated in diabetic rats, and TE suppressed both of them, whereas TE showed no effect on the polyol pathway metabolites. The results indicate that transition metals play a significant role in the formation of MG and 3-DG via oxidative stress and SSAO activity. PMID- 16260350 TI - Increased interleukin-18 content and angiogenic activity of sera from diabetic (Type 2) patients with background retinopathy. AB - Hyperglycaemia increases inflammatory cytokine concentration in the blood. Elevated levels of interleukin-18 (IL-18), a cytokine belonging to the interleukin-1 (IL-1) family, were recently reported in patients with Type 2 diabetes mellitus (DM2) and nephropathy. The aim of the present work was an examination of IL-18 concentration in the sera of elderly DM2 patients with nonproliferative retinopathy and age-matched control people and an estimation whether this cytokine plays pro- or anti-angiogenic role in in vivo angiogenic activity of their sera in mice cutaneous angiogenesis test. Recombinant human IL 18 injected intradermally to murine skin induced significant neovascular reaction. DM2 patients sera contained higher concentration of IL-18 and induced stronger neovascular reaction in mice skin than did the sera of corresponding control people. Sera from both groups of people after neutralization with antihuman IL-18 antibodies lost substantial part of their angiogenic activity. PMID- 16260351 TI - Early treadmill exercise decreases intrastriatal hemorrhage-induced neuronal cell death and increases cell proliferation in the dentate gyrus of streptozotocin induced hyperglycemic rats. AB - Intracerebral hemorrhage (ICH) is a severe complication in diabetic patients. Currently, physical exercise is recommended as a behavioral intervention to promote functional recovery in brain diseases, including ICH. Recently, hyperglycemia is known to aggravate brain injury in experimental ICH. Here, we examined the effect of treadmill exercise on the intrastriatal hemorrhage-induced neuronal cell death and cell proliferation in the dentate gyrus of hyperglycemic rats. Hyperglycemia was induced by the intraperitoneal injection of 50 mg/kg streptozotocin (STZ). Intrastriatal hemorrhage was induced by the infusion of 0.2 U collagenase into the striatum using stereotaxic instrument. Rats in the exercise groups were forced to run on a treadmill for 30 min daily for 10 days. Apoptosis was assessed by the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. Cell proliferation was assessed by the 5-bromo 2'-deoxyuridine (BrdU) immunohistochemistry. Our data showed that in rats started treadmill exercise 24 h after ICH induction, the size of lesion induced by hemorrhage and the number of apoptotic cells were decreased significantly. The number of proliferating cells in the dentate gyrus was significantly decreased in hyperglycemic rats. Treadmill exercise markedly enhanced cell proliferation in the dentate gyrus of hyperglycemic rats. The data suggest that treadmill exercise may provide therapeutic value to ICH patients with hyperglycemia by suppressing neuronal apoptosis and increasing cell proliferation. PMID- 16260352 TI - High serum TNF-alpha level in Type 2 diabetic patients with microangiopathy is associated with eNOS down-regulation and apoptosis in endothelial cells. AB - A high dose of tumor necrosis factor (TNF)-alpha induces endothelial dysfunction and enhances apoptosis in vitro. The present study was conducted to examine whether incubating human umbilical vein endothelial cells (HUVECs) with serum from Type 2 diabetic patients complicated with retinopathy and/or microalbuminemia demonstrate endothelial dysfunction. Serum levels of TNF-alpha and vascular endothelial growth factor (VEGF) were elevated in diabetic patients. Plasma levels of TNF-alpha, two soluble TNF-alpha receptors (sTNFR), and VEGF were assessed in diabetic patients (CD, n=21) complicated with retinopathy and/or nephropathy, uncomplicated diabetic patients (UD, n=18), and in healthy normal participants (NS, n=16). In HUVECs incubated with patient's serum, endothelial constitutive nitric oxide synthase (eNOS) protein expressions were measured by Western blot analysis. Apoptosis in HUVECs was determined by optical microscopy, DNA fragmentation, and CPP32-like protease activity. Serum TNF-alpha, sTNFR-I, and asymmetric dimethylarginine (ADMA), an endogenous inhibitor of NOS, in CD were significantly higher than in UD or NS. While, serum sTNFR-I and VEGF levels were significantly increased in the both diabetic patients, compared with those of NS, no difference was observed in the serum TNF-alpha, sTNFR-II, and ADMA levels between UD and NS. eNOS down-regulation and apoptosis were seen in HUVECs incubated with serum from CD for 24 h, but those observations were completely counteracted in the incubation by the addition of the antihuman TNF-alpha antibody. These results imply that eNOS down-regulation in CD is associated with high serum TNF-alpha levels despite of high serum of VEGF levels. Therefore, endothelial dysfunction in diabetic patients complicated with microangiopathy may, in part, be attributed to high serum TNF-alpha levels. PMID- 16260353 TI - Effect of the administration of a single dose of nateglinide on insulin secretion at two different concentrations of glucose in healthy individuals. AB - BACKGROUND: Nateglinide is a D-phenylalanine derivative that stimulates fast insulin secretion with a short activity span. It has been suggested that the hypoglycemic effect of nateglinide is related to the glucose concentration, an aspect that still has not been completely evaluated in human beings. OBJECTIVE: The aim of this study is to evaluate the effect of nateglinide on the insulin secretion at two different concentrations of glucose level. PARTICIPANTS AND METHODS: A randomized, double-blind, cross-over, placebo-controlled clinical trial with two parallel groups was carried out; each group was made up by six healthy volunteers who were submitted to a hyperglycemic-hyperinsulinemic clamp technique on two different occasions, one of them prior to the administration of 120 mg nateglinide and the other one prior to the administration of an homologated placebo. One group was submitted to and maintained at a hyperglycemia of 6.9 mmol/l above the fasting glucose level and the other group at a hyperglycemia of 4.1 mmol/l above the baseline of fasting glucose level. RESULTS: In volunteers submitted to the clamp at 4.1 mmol/l above the baseline of glucose level, the insulin secretion in the early phase was 212.4+/-55.8 pmol/l in the placebo test versus 338.4+/-124.8 pmol/l in the nateglinide test (P<.05), whereas in the group submitted at 6.9 mmol/l over the baseline, no significant differences were observed. CONCLUSION: Nateglinide increased the early insulin secretion in healthy individuals submitted to a mild hyperglycemia, but not at high glucose concentrations. PMID- 16260354 TI - Myocardial injury with biomarker elevation in diabetic ketoacidosis. AB - We report of two patients with severe ketoacidosis, minute elevations of myocardial biomarkers (troponin T and CK-MB) and initial ECG changes compatible with myocardial infarction (MI). All successive investigations, including coronary arteriography, were normal, and the patients recovered fully without further evidence of ischemic heart disease. We suggest that acidosis and very high levels of free fatty acids could cause membrane instability and biomarker leakage. Regardless of the pathogenesis, these two case stories suggest that nonspecific myocardial injury may occur in severe diabetic ketoacidosis and that the presence of minute biomarker elevation and ECG changes does not necessarily signify MI. PMID- 16260355 TI - Bony palatal necrosis in a diabetic patient secondary to palatal rotational flap. AB - Donor site necrosis is a very rare complication of palatal rotational flaps. The aim of this case report is to present a 43-year-old female, Type I diabetic patient with a chronic oroantral fistula in the right second molar region. The patient had bony necrosis in the donor site following palatal rotational flap operation. The treatment approach and the alternative methods are discussed. PMID- 16260356 TI - Perceptions of predisposing and protective factors for perinatal depression in same-sex parents. AB - Increasing numbers of women are choosing to have children in the context of same sex relationships or as "out" lesbian or bisexual individuals. This study used qualitative methods to assess perceived predisposing and protective factors for perinatal depression in lesbian, gay, bisexual, and queer (LGBQ) women. Two focus groups with LGBQ women were conducted: 1) biological parents of young children and 2) nonbiological parents of young children or whose partners were currently pregnant. Three major themes emerged. Issues related to social support were primary, particularly related to disappointment with the lack of support provided by members of the family of origin. Participants also described issues related to the couple relationship, such as challenges in negotiating parenting roles. Finally, legal and policy barriers (e.g., second parent adoption) were identified as a significant source of stress during the transition to parenthood. Both lack of social support and relationship problems have previously been identified as risk factors for perinatal depression in heterosexual women, and legal and policy barriers may represent a unique risk factor for this population. Therefore, additional study of perinatal mental health among LGBQ women is warranted. PMID- 16260357 TI - Is collaborative practice a malpractice risk? Myth versus reality. PMID- 16260359 TI - Closed claims analysis of 65 medical malpractice cases involving nurse-midwives. AB - The threat of litigation in clinical midwifery is evident in daily practice. Although midwives have not had the same risk of claims and suits as obstetricians, all obstetric providers are potentially subject to claims of malpractice. Closed claims analysis has been used in the past to review risk patterns and to heighten awareness of certain risks. It is a methodology that can suggest corrective or preventive action for future practice and thereby minimize the risk of future errors. An analysis of nurse-midwifery closed claims can help to evaluate past risk and proactively modify future liability. PMID- 16260360 TI - Improving midwifery practice: the American College of Nurse-Midwives' benchmarking project. AB - Quality management in clinical practice involves the use of numerous techniques that monitor the quality of care clinicians provide. Quality improvement is an approach to quality management that emphasizes system and processes, rather than a focus on individual performance. Quality improvement examines objective data to improve these processes, even when high standards of performance appear to have been met. Benchmarking measures one's processes and outcomes against "best in class" and is a part of a quality improvement program. By using benchmarking to provide goals for realistic process improvement and identification of the most efficient and effective methods of meeting all of their customer's needs, health care providers can document their effectiveness in terms of cost, quality, and satisfaction. This article details the American College of Nurse-Midwives' benchmarking project and presents benchmarks for obstetric practice from the year 2004. PMID- 16260361 TI - Liability in triage: management of EMTALA regulations and common obstetric risks. AB - The Emergency Medical Treatment and Active Labor Act (EMTALA) affects all clinicians who provide triage care for pregnant women. EMTALA has specific regulations for hospitals relative to women in active labor. Violations can carry stiff penalties. It is critical for clinicians performing obstetric triage to understand the duties and obligations of this law. This article discusses EMTALA and reviews common liability risks in obstetric triage as well as strategies to modify those risks. PMID- 16260362 TI - Trouble in the office: liability issues in ambulatory care. AB - Most clinicians are keenly aware of the liability risks associated with adverse birth outcomes for mother and/or infant. This level of awareness and concern also needs to extend to the day-to-day management of office practice. Clinicians who care for healthy women and their families may see abnormalities infrequently. The low yield of abnormal results means that close monitoring is essential to prevent overlooking the rare abnormal finding. This article focuses on three areas of liability risk for midwives and others who practice in the ambulatory setting: 1) inadequate tracking of test and referral results, 2) failure to diagnose, and 3) poor communication. Although none of these issues are limited to the ambulatory setting, only office practice is addressed in this article. PMID- 16260363 TI - Shoulder dystocia: etiology, common risk factors, and management. AB - Shoulder dystocia and brachial plexus injury occur in 0.5% to 1.5% of all births. Risk factors for both include maternal obesity, excessive prenatal weight gain, maternal diabetes, protracted labor, and fetal macrosomia. These factors are involved in only about 50% of births complicated by shoulder dystocia or brachial plexus injury. Shoulder dystocia has a low recurrence rate (9.8%-16.7%), although history of previous shoulder dystocia is the most reliable predictor of occurrence. Brachial plexus injury is the most common morbidity associated with shoulder dystocia, but 50% of newborns who present with this injury were not subject to shoulder dystocia at birth. Most brachial plexus injuries are transient, although 5% to 22% become permanent. Shoulder dystocia followed by permanent brachial plexus injury or mental impairment is one of the leading causes of malpractice allegations. Prompt assessment and management of shoulder dystocia and preparation to maximize the efficiency of shoulder dystocia maneuvers are critical. Documentation of the appropriate use of maneuvers to relieve shoulder dystocia demonstrates standard of care practice, thereby decreasing the potential for successful malpractice allegations. PMID- 16260364 TI - Intrauterine asphyxia: clinical implications for providers of intrapartum care. AB - Advances in science and technology have allowed researchers to gain a better understanding of the pathophysiology leading to long-term neurologic damage in newborns. Intrapartum events are now known to be an infrequent cause of adverse neurologic outcome. Clinicians caring for women during labor must have an understanding of the pathophysiology of intrauterine asphyxia as well as an awareness of the capabilities and limitations of available intrapartum fetal assessment tools to diagnose intrauterine fetal asphyxia or predict neurologic outcome. This article reviews the physiology of acid-base balance and fetal gas exchange as well as the current scientific understanding of the role of intrauterine asphyxia in the pathophysiology of neonatal encephalopathy and cerebral palsy. Recommendations for care and documentation are included. PMID- 16260365 TI - System errors in intrapartum electronic fetal monitoring: a case review. AB - Intrapartum electronic fetal monitoring (EFM) interpretation and management continue to be a common issue in litigation involving adverse outcomes in term pregnancies. This article uses a case study approach to illustrate system errors related to intrapartum EFM. Common system errors related to use of intrapartum EFM include knowledge deficits, communication failures, and fear of conflict. Strategies for reducing error and the promotion of a patient safety approach to risk management in EFM are discussed, with an emphasis on the importance of a true team approach to EFM education, interpretation, and management. PMID- 16260367 TI - The National Practitioner Data Bank: information for and about midwifery. AB - The National Practitioner Data Bank (NPDB), mandated by federal legislation in 1986, serves as a mechanism to protect the public from unsafe practitioners who attempt to avoid discovery of prior negligent behavior or malpractice record(s) by moving from state to state. Reporting to the NPDB about malpractice payments on behalf of nurse-midwives began in 1990. Reporting of providers excluded from Medicare and Medicaid program participation began in September 1999. Practitioners who were already in an excluded status at that time were reported. Reports of adverse action against a nurse-midwife can be submitted to the NPDB by a state licensure board, a governmental agency, hospitals, health maintenance organizations, or other health care organizations. Reporting of licensing actions and clinical privilege actions are not required, although these may be voluntarily reported. The NPDB received 484 reports about nurse-midwives from September 1, 1999, to March 31, 2005. Of the 484 reports, 375 have an obstetric malpractice code. The median claim payment made on behalf of nurse-midwives during this period is 225,000 dollars. Although limited, the NPDB is the only systematic national source of nurse-midwifery malpractice data collection. PMID- 16260366 TI - Use of the vacuum extractor by midwives--what has changed in the last decade? AB - This article provides an update on use of vacuum extraction by certified nurse midwives (CNMs) and certified midwives (CMs). Research findings from the last decade relevant to the training and use of vacuum extractors are reviewed along with professional guidelines and legal implications. PMID- 16260368 TI - Liability concerns: a view from the American College of Nurse-Midwives. AB - The national office of the American College of Nurse-Midwives (ACNM) is the center of day-to-day administration for the professional organization. Staff members respond to requests for information and assistance from certified nurse midwives or certified midwives and facilitate the work of the Board, Divisions, and Committees. This article reviews the questions most frequently asked of the Professional Services department about risk management and professional liability issues and summarizes the most important lessons learned from the calls received. The focus is on 5 topic areas: 1) collaboration confusion, 2) systems needed to implement and effectively track diagnostic tests and referrals, 3) expansion of services beyond the core competencies, 4) patient satisfaction and patient complaints, and 5) maintaining medical malpractice coverage. PMID- 16260369 TI - Parvovirus B19 in pregnancy. AB - Pregnant women who report possible exposure to a viral illness can present a clinical predicament for obstetric providers. Although some viruses are benign in pregnancy, others can have serious consequences. Parvovirus B19, the causative agent of fifth disease, is one of the more serious of the common viruses. Understanding the pathophysiology and appropriate testing for parvovirus will help providers better counsel their patients. PMID- 16260370 TI - Patient safety Internet resources for midwifery practice. PMID- 16260376 TI - Detection of chlorinated DNA and RNA nucleosides by HPLC coupled to tandem mass spectrometry as potential biomarkers of inflammation. AB - Upon inflammation, activated neutrophils secrete myeloperoxidase, an enzyme able to generate hypochlorous acid (HOCl) from hydrogen peroxide and chloride ions. An analytical method, involving HPLC coupled to electrospray tandem mass spectrometry, has been set-up to detect low levels of HOCl-induced nucleic acids lesions, including both ribo and 2'-deoxyribonucleoside derivatives of 8 chloroguanine, 8-chloroadenine and 5-chlorocytosine. Validation of the developed method was achieved using isolated cells treated with HOCl. The method was found to be sensitive enough to allow the measurement of background levels of 5-chloro 2'-deoxycytidine in the DNA of human white blood cells isolated from 7 mL of blood. PMID- 16260377 TI - Porous graphitic carbon chromatography-tandem mass spectrometry for the study of isoprostanes in human cerebrospinal fluid. AB - F2-isoprostanes are produced by the non-enzymatic peroxidation of arachidonic acid in membrane phospholipids. This paper describes a new method for the determination of all four classes of F2-isoprostanes in human cerebrospinal fluid (CSF) involving separation on a 1 mm x 150 mm porous graphitic carbon (PGC) column and detection by triple quadrupole mass spectrometry in negative-ion electrospray mode. The sample pre-treatment consisted of an ultrafiltration step, following which 300 microl of CSF sample could be injected directly onto a 1 mm x 10 mm PGC guard column functioning as a trap for the analytes. The loading solvent was Milli-Q water at 125 microl/min. After 3 min, the sample was switched into the separation column. The F2-isoprostanes were separated in 20 min using a linear solvent gradient comprising water, methanol, acetonitrile and ammonium hydroxide at a pH of 9.5 and a flow of 50 microl/min The limit of detection (calculated as 3S/N) was approximately 40 pM (14 pg/ml). The assay was linear within the examined range (18-450 pg/ml), using CSF spiked with iPF2alpha-III standard (r(2)>0.995). Repeatability data were calculated for CSF spiked to 90 pg/ml and the relative standard deviation (RSD) obtained was 3% (n=6). PMID- 16260378 TI - Assessment of in situ cellular glutathione labeling with naphthalene-2,3 dicarboxaldehyde using high-performance liquid chromatography. AB - We have presently studied a dialdehydic reagent, i.e. naphthalene-2,3 dicarboxaldehyde (NDA), as a fluorogenic probe for the labeling of intracellular reduced glutathione (GSH), using a yeast strain Candida albicans as a cell model. Chemical reactivity of NDA with both amino and sulfhydryl groups of the GSH molecule leads to a highly selective detection. Moreover, fluorescence properties of the resulting adduct fit well with most of modern instruments adapted for in situ measurements, and equipped with an argon laser. After incubation of cells with 100 microM of NDA for 20 min, cells were harvested and corresponding lysates obtained after a freezing cycle, were suspended in 0.2M borate buffer pH 9.2 and analysed with HPLC (column: Spherisorb ODS-2 (125 mm x 4.6 mm i.d.) 5 microm; mobile phase: methanol-0.01 M phosphate buffer pH 6.5 (20:80, v/v) at a flow rate of 0.8 mL min(-1); spectrofluorimetric detection: lambda(exc)=430 nm and lambda(em)=530 nm). The GSH-NDA adduct was identified in the yeast strain extracts using the reported HPLC technique and quantified versus a calibration curve of NDA derivatized with an excess of GSH (linearity range: 9-230 nM). The cell loading step of the free probe NDA and the extraction efficiency of the resulting NDA-GSH adduct were optimized. PMID- 16260379 TI - Determination of urinary 8-hydroxy-2'-deoxyguanosine by two approaches-capillary electrophoresis and GC/MS: an assay for in vivo oxidative DNA damage in cancer patients. AB - 8-Hydroxy-2'-deoxyguanosine (8OHdG) has been considered as an excellent marker of oxidative DNA damage associated with age-related diseases such as cancer. In this paper, two sensitive methods-capillary electrophoresis with electrochemical detection (CE-ECD) and gas chromatography/mass spectrometry (GC/MS) were developed for urinary 8OHdG analysis. The R.S.D. of the spiked recovery of the two methods for determining urinary 8OHdG was 4.03% and 8.25%, respectively, and the results from the two methods have a good consistency (r=0.999, P<0.01). The developed CE-ECD method was applied to investigate the urinary 8OHdG levels in different cancer patients and follow up the response of therapy. It was found that the excretion levels of urinary 8OHdG in cancer patients were significantly higher than those in healthy persons (35.26+/-27.96 nM versus 13.51+/-5.08 nM, P<0.05), and cancer patients receiving surgical therapy and chemotherapy showed a significant decrease in urinary 8OHdG. PMID- 16260380 TI - Suppression of murine cerebral F2-isoprostanes and F4-neuroprostanes from excitotoxicity and innate immune response in vivo by alpha- or gamma-tocopherol. AB - Oxidative damage to brain is a featured shared by several destructive and degenerative diseases and is thought to contribute to disease pathogenesis. Two commonly proposed sources of the increased free radical stress that leads to oxidative damage in several of these diseases are excitotoxicity and activation of innate immunity, both of which are proposed pharmacologic targets. Here we used models of excitotoxicity, intracerebroventricular (ICV) kainate (KA), and innate immune activation, ICV lipopolysaccharide (LPS), to test the effectiveness of peripherally administered alpha-tocopherol (AT) and gamma-tocopherol (GT) as neuroprotectants. We quantified murine cerebral oxidative damage by measuring F(2)-isoprostanes (IsoPs) and F(4)-neuroprostanes (NeuroPs) using stable isotope dilution methods followed by gas chromatography-mass spectrometry with selective ion monitoring. Our data showed that peripherally administered AT and GT were equally effective at suppressing acute oxidative damage from direct excitotoxicity caused by KA. In contrast, peripherally administered AT, but not GT, was effective at suppressing delayed neuronal oxidative damage from activated glial innate immune response. These data imply that AT may be more broadly protective of cerebrum from oxidative damage in different disease contexts. PMID- 16260381 TI - Oxidative stress and DNA damage caused by the urban air pollutant 3-NBA and its isomer 2-NBA in human lung cells analyzed with three independent methods. AB - The air pollutant 3-nitrobenzanthrone (3-NBA), emitted in diesel exhaust, is a potent mutagen and genotoxin. 3-NBA can isomerise to 2-nitrobenzanthrone (2-NBA), which can become more than 70-fold higher in concentration in ambient air. In this study, three independent methods have been employed to evaluate the oxidative stress and genotoxicity of 2-NBA compared to 3-NBA in the human A549 lung cell line. HPLC-EC/UV was applied for measurements of oxidative damage in the form of 8-oxo-2'-deoxyguanosine (8-oxodG), (32)P-HPLC for measurements of lipophilic DNA-adducts, and the Comet assay to measure a variety of DNA lesions, including oxidative stress. No significant oxidative damage from either isomer was found regarding formation of 8-oxodG analysed using HPLC-EC/UV. However, the Comet assay (with FPG-treatment), which is more sensitive and detects more types of damages compared to HPLC-EC/UV, showed a significant effect from both 3-NBA and 2-NBA. (32)P-HPLC revealed a strong DNA-adduct formation from both 3-NBA and 2-NBA, and also a significant difference between both isomers compared to negative control. These results clearly show that 2-NBA has a genotoxic potential. Even if the DNA-adduct forming capacity and the amount of DNA lesions measured with the (32)P-HPLC and Comet assay is about one third of 3-NBA, the high abundance of 2-NBA in ambient air calls for further investigation and evaluation of its health hazard. PMID- 16260382 TI - Determination of 3-nitrotyrosine in human urine at the basal state by gas chromatography-tandem mass spectrometry and evaluation of the excretion after oral intake. AB - 3-Nitrotyrosine (NO(2)Tyr) is a potential biomarker of reactive-nitrogen species (RNS) including peroxynitrite. 3-Nitrotyrosine occurs in human plasma in its free and protein-associated forms and is excreted in the urine. Measurement of 3 nitrotyrosine in human plasma is invasive and associated with numerous methodological problems. Recently, we have described an accurate method based on gas chromatography (GC)-tandem mass spectrometry (MS) for circulating 3 nitrotyrosine. The present article describes the extension of this method to urinary 3-nitrotyrosine. The method involves separation of urinary 3 nitrotyrosine from nitrite, nitrate and l-tyrosine by HPLC, preparation of the n propyl-pentafluoropropionyltrimethylsilyl ether derivatives of endogenous 3 nitrotyrosine and the internal standard 3-nitro-l-[(2)H(3)]tyrosine, and GC tandem MS quantification in the selected-reaction monitoring mode under negative ion chemical ionization conditions. In urine of ten apparently healthy volunteers (years of age, 36.5+/-7.2) 3-nitrotyrosine levels were determined to be 8.4+/ 10.4 nM (range, 1.6-33.2 nM) or 0.46+/-0.49 nmol/mmol creatinine (range, 0.05 1.30 nmol/mmol creatinine). The present GC-tandem MS method provides accurate values of 3-nitrotyrosine in human urine at the basal state. After oral intake of 3-nitro-l-tyrosine by a healthy volunteer (27.6 microg/kg body weight) 3-nitro-l tyrosine appeared rapidly in the urine and was excreted following a biphasic pharmacokinetic profile. Approximately one third of administered 3-nitro-l tyrosine was excreted within the first 8 h. The suitability of the non-invasive measurement of urinary 3-nitrotyrosine as a method of assessment of oxidative stress in humans remains to be established. PMID- 16260383 TI - The AIDS dementia complex: clinical and basic neuroscience with implications for novel molecular therapies. AB - The AIDS dementia complex (ADC, also referred to as HIV-associated cognitive impairment) is a common disorder among HIV-infected patients associated with both inflammatory and neurodegenerative processes. This review describes recent advances in the clinical and basic neurosciences of HIV infection and discusses the multivariable nature of what has become a chronic disorder in the context of highly active antiretroviral therapies (HAART). Since its initial description twenty years ago, advances in cell and molecular biology along with those in neuroimaging have furthered our understanding of the underlying pathogenic mechanisms. The clinical and neuropsychological profile of ADC is generally consistent with a "frontal-subcortical" pattern of injury. Neuropathogenesis is largely driven by indirect mechanisms mediated by infected, or more commonly, immune activated macrophages, which secrete viral and host-derived factors. Magnetic resonance spectroscopy (MRS) provides a robust in vivo method to measure the inflammatory and neurotoxic events triggered by these factors and their associated signals. Although the use of combined or highly active antiretroviral therapies (HAART) has significantly improved survival rates, cerebral injury and cognitive impairment remain common events. Factors such as aging and chronic infection will likely impact the course of this disease, its pathogenesis, and treatment. The combined observations presented in this review suggest a number of critical areas for future inquiry. PMID- 16260385 TI - The shifting patterns of HIV encephalitis neuropathology. AB - HIV infected macrophages infiltrate the nervous system early in the progression of HIV infection, leading to a complex set of neuropathological alterations including HIV encephalitis (HIVE), leukoencephalopathy and vacuolar myelopathy that in turn result in neurodegeneration of selective cellular populations and pathways involved in regulating cognitive and motor functioning. Rapid progress in the development of highly active antiretroviral therapy (HAART) has changed the patterns of HIV related neuropathology and neurological manifestations in the past 10 years. The prevalence of opportunistic infections and central nervous system (CNS) neoplasms has decreased, and some groups have proposed that the frequency of chronic forms of HIVE have been rising as the HAART-treated HIV population ages. Accordingly, clinical manifestations have shifted from severe dementia forms to more subtle minor cognitive impairment, leading to the suggestion of a classification of HIV associated neurological conditions into an inactive form, a chronic variety, and a 'transformed' variant. From a neuropathological point of view these variants might correspond to: a) aggressive forms with severe HIVE and white matter injury, b) extensive perivascular lymphocytic infiltration, c) 'burnt-out' forms of HIVE and d) aging-associated amyloid accumulation with Alzheimer's-like neuropathology. Factors contributing to the emergence of these variants of HIVE include the development of viral resistance, immune reconstitution, anti-retroviral drug toxicity and co-morbid factors (e.g., methamphetamine, HCV). More detailed characterization of these proposed variants of HIVE is important in order to better understand the pathogenesis of HIV-associated neurological damage and to design more effective treatments to protect the nervous system. PMID- 16260387 TI - Human brain derived cell culture models of HIV-1 infection. AB - During the clinical course of acquired immune deficiency syndrome, infection of the CNS by human immunodeficiency virus-1 (HIV-1) may ultimately result in the impairment of cognitive, behavioral and motor functions. Viral neuropathogenesis involves inflammatory molecules and neurotoxins produced from infected and immune activated lymphocytes, microglial cells and astrocytes. Here, we discuss the current understanding of HIV-1 infection of the CNS and various cell culture systems from the developing human brain in order to study the neurobiology of HIV 1 infection, the mechanisms contributing to HIV-1 infection, and disease progression. PMID- 16260384 TI - Mononuclear phagocytes in the pathogenesis of neurodegenerative diseases. AB - Brain mononuclear phagocytes (MP, bone marrow monocyte-derived macrophages, perivascular macrophages, and microglia) function to protect the nervous system by acting as debris scavengers, killers of microbial pathogens, and regulators of immune responses. MP are activated by a variety of environmental cues and such inflammatory responses elicit cell injury and death in the nervous system. MP immunoregulatory responses include secretion of neurotoxic factors, mobilization of adaptive immunity, and cell chemotaxis. This incites tissue remodelling and blood-brain barrier dysfunction. As disease progresses, MP secretions engage neighboring cells in a vicious cycle of autocrine and paracrine amplification of inflammation leading to tissue injury and ultimately destruction. Such pathogenic processes tilt the balance between the relative production of neurotrophic and neurotoxic factors and to disease progression. The ultimate effects that brain MP play in disease revolves "principally" around their roles in neurodegeneration. Importantly, common functions of brain MP in neuroimmunity link highly divergent diseases (for example, human immunodeficiency virus type-one associated dementia, Alzheimer's disease and Parkinson's disease). Research into this process from our own laboratories and those of others seek to harness MP inflammatory processes with the intent of developing therapeutic interventions that block neurodegenerative processes and improve the quality of life in affected people. PMID- 16260388 TI - Rodent model systems for studies of HIV-1 associated dementia. AB - Understanding of HIV-1 neuropathogenesis and development of rationale therapeutic approaches requires relevant animal models. The putative mechanisms of neuroinflammatory and neurotoxic events triggered by HIV-1 brain infection are reflected by a number of rodent models. These include transgenic animals (either expressing viral proteins or pro-inflammatory factors), infection with murine retroviruses, and severe combined immunodeficient (SCID) mice reconstituted with human lymphocytes and injected intracerebrally with HIV-1-infected human monocyte derived macrophages. The potential importance and limitations of the models in reflecting human disease are discussed with emphasis on their utility for development of therapies to combat HIV-1-associated neurologic impairment. PMID- 16260386 TI - Molecular targets of opiate drug abuse in neuroAIDS. AB - Opiate drug abuse, through selective actions at mu-opioid receptors (MOR), exacerbates the pathogenesis of human immunodeficiency virus-1 (HIV-1) in the CNS by disrupting glial homeostasis, increasing inflammation, and decreasing the threshold for pro-apoptotic events in neurons. Neurons are affected directly and indirectly by opiate-HIV interactions. Although most opiates drugs have some affinity for kappa (KOR) and/or delta (DOR) opioid receptors, their neurotoxic effects are largely mediated through MOR. Besides direct actions on the neurons themselves, opiates directly affect MOR-expressing astrocytes and microglia. Because of their broad-reaching actions in glia, opiate abuse causes widespread metabolic derangement, inflammation, and the disruption of neuron-glial relationships, which likely contribute to neuronal dysfunction, death, and HIV encephalitis. In addition to direct actions on neural cells, opioids modulate inflammation and disrupt normal intercellular interactions among immunocytes (macrophages and lymphocytes), which on balance further promote neuronal dysfunction and death. The neural pathways involved in opiate enhancement of HIV induced inflammation and cell death, appear to involve MOR activation with downstream effects through PI3-kinase/Akt and/or MAPK signaling, which suggests possible targets for therapeutic intervention in neuroAIDS. PMID- 16260389 TI - The role of monocytes and perivascular macrophages in HIV and SIV neuropathogenesis: information from non-human primate models. AB - Perivascular macrophages are located in the perivascular space of cerebral microvessels and thus uniquely situated at the intersection between the brain parenchyma and blood. Connections between the nervous and immune systems are mediated in part through these cells that are ideally located to sense perturbations in the periphery and turnover by cells entering the central nervous system (CNS) from the circulation. It has become clear that unique subsets of brain macrophages exist in normal and SIV- or HIV-infected brains, and perivascular macrophages and similar cells in the meninges and choroid plexus play a central role in lentiviral neuropathogenesis. Common to all these cell populations is their likely replacement within the CNS by monocytes. Studies of SIV-infected non-human primates and HIV-infected humans underscore the importance of virus-infected and activated monocytes, which traffic to the CNS from blood to become perivascular macrophages, potentially drive the blood-brain barrier damage and cause neuronal injury. This review summarizes what we know about SIV- and HIV induced neuropathogenesis focusing on brain perivascular macrophages and their precursors in blood that may mediate HIV CNS infection and injury. PMID- 16260392 TI - HIV-1, chemokines and neurogenesis. AB - HIV-1 infection of the brain results in a large number of behavioural defecits accompanied by diverse neuropathological signs. However,it is not clear how the virus produces these effects or exactly how the neuropathology and behavioural defecits are related. In this article we discuss the possibility that HIV-1 infection may negatively impact the process of neurogenesis in the adult brain and that this may contribute to HIV-1 related effects on the nervous system. We have previously demonstrated that the development of the dentate gyrus during embryogenesis requires signaling by the chemokine SDF-1 via its receptor CXCR4. We demonstrated that neural progenitor cells that give rise to dentate granule neurons express CXCR4 and other chemokine receptors and migrate into the nascent dentate gyrus along SDF-1 gradients. Animals deficient in CXCR4 receptors exhibit a malformed dentate gyrus in which the migration of neural progenitors is stalled. In the adult, neurogenesis continues in the dentate gyrus. Adult neural progenitor cells existing in the subgranlar zone, that produce granule neurons, express CXCR4 and other chemokine receptors, and granule neurons express SDF-1 suggesting that SDF-1/CXCR4 signaling is also important in adult neurogenesis. Because the cellular receptors for HIV-1 include chemokine receptors such as CXCR4 and CCR5 it is possible that the virus may interfere with SDF-1/CXCR4 signaling in the brain including disruption of the formation of new granule neurons in the adult brain. PMID- 16260393 TI - Adjunctive therapies for HIV-1 associated neurologic disease. AB - In the past decade we have seen a milder phenotype and decreased incidence of HIV 1 associated dementia (HAD), largely due to the widespread use of combination chemotherapy to reduce viral burden. However, the prevalence of neurologic disease in people living with HIV-1 has actually increased, raising significant concerns that new therapeutic strategies, directed at restoring neuronal and glial homeostasis and signaling in the central nervous system (CNS), as opposed to directly interfering with the life cycle of HIV-1, must be developed. In this review, we focus briefly on previous Phase 1 clinical trials for adjunctive (i.e., chemotherapeutic agents that do not have a primary antiretroviral mechanism of action) therapy in patients with HAD, followed by an overview of key molecular events in the neuropathogenesis of HAD, and then discuss in more detail our rationale for investigating the effects of therapeutic agents that restore impaired mitochondrial bioenergetics in the CNS. Specifically, we focus on agents that either work in part through K-ATP channels, present in both mitochondria and plasma membranes, and agents that work to weakly uncouple the respiratory capacity of the electron transport chain in mitochondria from ATP production. We propose these agents may be complementary to currently available antiretroviral agents and may significantly improve the capacity of CNS infected with HIV-1 to meet increased bioenergetic demands involved in normal synaptic communication. PMID- 16260391 TI - The signaling and apoptotic effects of TNF-related apoptosis-inducing ligand in HIV-1 associated dementia. AB - HIV-1 Associated Dementia (HAD) develops during progressive HIV-1 infection and is characterized by cognitive impairments, behavioral disorders and potential progressive motor abnormality. Abnormal inflammation within the central nervous system (CNS), activation of macrophage/microglia and involvement of proinflammatory cytokines have been suggested as primary factors in the pathogenesis of HAD. Impairment of neuronal function and neuronal cell death are believed to be the end pathophysiological result of HAD. TNF-related apoptosis inducing ligand (TRAIL), a member of the TNF family of cytokines, was suggested to participate in apoptotic cell death during HAD. As a death ligand, TRAIL was originally thought to target only tumor cells. TRAIL is not typically present in CNS; however, emerging data show that TRAIL can be induced by immune stimuli on macrophage and microglia, major disease effector cells during HAD. Upregulated TRAIL may then cause neuronal apoptosis through direct interaction with TRAIL receptors on neurons or through macrophage death-mediated release of neurotoxins. In this review, we summarize the pivotal role of TRAIL in HAD and TRAIL-initiated intracellular death cascades that culminate in neuronal apoptosis as observed in HAD. PMID- 16260390 TI - Molecular and cellular mechanisms of neuronal cell death in HIV dementia. AB - The deaths of neurons, astrocytes and endothelial cells have been described in patients with HIV (human immunodeficiency virus) dementia. HIV-1 does not infect neurons; instead, neurotoxic substances shed by infected glia and macrophages can induce a form of programmed cell death called apoptosis in neurons. These neurotoxins include the HIV-1 proteins Tat and gp120, as well as pro-inflammatory cytokines, chemokines, excitotoxins and proteases. In this article we review the evidence for apoptosis of various cell types within the brain of HIV-infected patients, and describe in vitro and in vivo experimental studies that have elucidated the mechanisms by which HIV causes apoptosis of brain cells. PMID- 16260396 TI - Introduction: aging education in a global context. AB - Introduction to a collection of articles on aging education in a global context based on the proceedings of the annual conference of the Association for Gerontology in Higher Education (AGHE). The volume includes a dual focus on (1) issues related to gerontology and geriatric education around the world and (2) issues related to teaching about global and cross-cultural aging. Several of the chapters provide useful information and resources for educators seeking ways to incorporate cross-cultural aging into their courses. The remaining three chapters deal with issues related to aging education in Kenya, Japan and China. PMID- 16260397 TI - Globalization and the experiences of aging. AB - Globalization is a product of urbanization and economic intensification which has escalated since the 1970s. Globalized markets have created many of the features of modern life including consumerism, increased cultural homogeneity, increased social polarization, erosion of the sovereignty of nation states, and delocalization of daily life. The consequences of globalization for older people are primarily disadvantageous. Most notably is the restructuring of the redistributive economy. The globalization of labor results in lower wages and marked social stratification. Also family life is altered with fewer relatives who must balance work and family obligations. PMID- 16260398 TI - Course design on aging: incorporating cross-cultural perspectives that challenge assumptions about assessment and service delivery. AB - This article explores the benefits of using cross-cultural, anthropological approaches to help students understand the varying experience of aging within the United States and around the globe. Through cultural comparison and increased knowledge of the cultural context of aging standardized aspects of practice such as assessment protocols and concepts of function are critiqued, so that students will be better prepared to work as professionals with elders from diverse cultural backgrounds. A gerontology course designed to accomplish these goals is used as the focus of discussion and as an example of how such a course can be constructed. PMID- 16260399 TI - Cross-cultural perspectives in thanatology: through a prism of religious faiths. AB - Recent decades have witnessed an increase in thanatology education in colleges and universities. However, the infusion into thanatology curricula of religious faiths as they affect behaviors, experiences and emotions of dying individuals and survivors is still in its infancy. In this article I describe an effective approach I have used to integrate various religious beliefs and practices into an undergraduate course on death and dying. Before presenting my approach, I provide a brief description of the state of death education in professional and undergraduate institutions in America and a rationale for infusing cross-cultural components into thanatology curricula. PMID- 16260400 TI - Heroes of their own stories: expressions of aging in international cinema. AB - This study of 14 international feature-length films (1988- 2003) is aimed at providing gerontologists with models of successful aging that portray elders as being valued within the context of community. Elders serve as role models and mentors for the young, and they resolve mid-life crises for the middle-aged. Elders complete their life's work in the context of community (aging in place), where they struggle to maintain abiding values and draws others to the community. The study concludes with examples of key visual metaphors utilized at the endings of selected films that affirm the role of elders as catalysts for change for the young, the middle-aged, and even their communities. PMID- 16260395 TI - Brain-derived neurotrophic factor as a prototype neuroprotective factor against HIV-1-associated neuronal degeneration. AB - Patients with human immunodeficiency virus type 1 (HIV-1) infection develop a broad spectrum of motor impairments and cognitive deficits, which follow or parallel cellular loss and atrophy in their brains. The viral envelope glycoprotein 120 (gp120) has been suggested to be a causal agent of neuronal loss. Therefore, reducing gp120 neurotoxicity may prevent neuronal degeneration seen in these patients. Here, we describe in vitro and in vivo experimental evidence that gp120 toxicity can be reduced by brain-derived neurotrophic factor (BDNF), a naturally occurring peptide that has been shown to block neurotoxin and trauma-induced neuronal injury. Moreover, we review the survival promoting properties of BDNF and the issues concerning its delivery into the brain, in an attempt to explain the rationale for exploring BDNF as a prototype trophic factor for a therapy to reduce neuronal cell death in HIV-1 infected patients. PMID- 16260401 TI - Teaching cross-cultural aging: using literary portrayals of elders from Chile and the United States. AB - Literary texts are cultural artifacts revealing a society's values and attitudes; reading literature about elders and old age can change readers' ageist attitudes. Beginning with these assumptions, I discuss ways of teaching cross-cultural aging in undergraduate literature courses, using Chilean texts paired with American texts. Students learn how old age is socially constructed and how writers can either reinforce or challenge negative societal stereotypes of elders. Chilean texts reveal Chileans' respect and affection by associating elders with nature and ascribing to them otherworldly wisdom. American texts respectfully depict elders connected to nature, but not as transcending the earthly. PMID- 16260394 TI - Experimental and potential future therapeutic approaches for HIV-1 associated dementia targeting receptors for chemokines, glutamate and erythropoietin. AB - Severe and debilitating neurological problems that include behavioral abnormalities, motor dysfunction and frank dementia can occur after infection with the human immunodeficiency virus-1 (HIV-1). Infected peripheral immune competent cells, in particular macrophages, infiltrate the central nervous system (CNS) and provoke a neuropathological response involving all cell types in the brain. HIV-1 infection results in activation of chemokine receptors, inflammatory mediators, extracellular matrix-degrading enzymes and glutamate receptor-mediated excitotoxicity, all of which can trigger numerous downstream signaling pathways that result in disruption of neuronal and glial function. Despite many major improvements in the control of viral infection in the periphery, a truly effective therapy for HIV-1 associated dementia is currently not available. This review will discuss experimental and potentially future therapeutic strategies based on recently uncovered pathologic mechanisms contributing to neuronal damage induced by HIV-1. PMID- 16260402 TI - Gerontology programs in Japanese higher education: brief history, current status, and future prospects. AB - The development of gerontological education is lagging behind in Japan in spite of Japan's large population of elders. Nevertheless, there are signs that this may be changing. In this paper we discuss how gerontology education has evolved in Japan over the past 40 years. Specifically, we provide an overview of the development of academic societies related to gerontology, the number of gerontological books published, government statements about gerontology education, and Japan's only graduate program in gerontology. We also identify some reasons for the delay in creating gerontology programs and propose specific steps that might be taken to reverse this trend. PMID- 16260403 TI - Gerontology education and research in Kenya: establishing a US-African partnership in aging. AB - This article reprises four presentations on Gerontology Education in Kenya, a seminar at the 2004 Annual Meeting of the Association of Gerontology in Higher Education. It describes the process by which the Gerontology Institute of Georgia State University established a 3-year gerontology education and research partnership with Kenyatta University in Nairobi, Kenya, and the field experiences of two scholars who have conducted aging research in Kenya. We provide four key elements of cultural competence and recommendations for American gerontologists wishing to establish international linkages. PMID- 16260405 TI - A comparision of the hemostatic effects of notoginseng and yun nan bai yao to placebo control. AB - INTRODUCTION: We previously demonstrated the hemostatic effects of notoginseng using a rat bleeding model. Yun-Nan-Bai-Yao is a proprietary product for external use in China to treat bleeding but the hemostatic effects have not been proven. This study was conducted to compare the hemostatic effects of notoginseng to that of Yun-Nan- Bai-Yao and placebo control. METHODS: Rats (n = 37) were randomized into 3 groups and their tails were transected 5mm from the tip in this blinded investigation. Group 1 received placebo (wheat flour, n = 17), group 2 received notoginseng (n = 10) and group 3 received the Yun-Nan-Bai-Yao (n = 10). The total bleeding time was determined and compared among groups. RESULTS: Beeding time in minutes was 29.7% and 22.3% lower in the notoginseng and Yun-Nan-Bai-Yao groups than the placebo group (p < 0.001 and p < 0.001, respectively). No significant difference was found between the two active groups (p = 0.418). CONCLUSIONS: When applied externally, both notoginseng and Yun-Nan-Bai-Yao provide appreciable hemostatic effects compared to placebo. PMID- 16260404 TI - Teaching Chinese health care professionals about community-based long-term care (CBLTC) in China. AB - Academic exchanges between the U.S. and other countries around the world are increasing and teaching students abroad is part of this trend. China is in its initial stage of developing gerontology education and is in great need of new concepts and ideas for dealing with its rapidly aging population. This paper discusses the challenges and rewards of teaching gerontology to health care professionals in China. To achieve the desired learning outcomes in another country requires culturally appropriate course materials and teaching methods; drawing on students' knowledge and expertise by using an interactive format and gaining students' respect. PMID- 16260406 TI - Hypoglycemic effect of ethanolic extract of Musa sapientum on alloxan induced diabetes mellitus in rats and its relation with antioxidant potential. AB - The antihyperglycemic effect of ethanolic extract of flowers of Musa sapientum (Musaceae), a herb (used in Indian folklore medicine for the treatment of diabetes mellitus) in alloxan induced diabetic rats. Oral administration of the ethanolic extract showed significant (p < 0.001) blood glucose lowering effect at 200 mg/kg in alloxan induced diabetic rats (120 mg/kg, i.p.) and the extract was also found to significantly (p < 0.001) scavenge oxygen free radicals, viz., superoxide dismutase (SOD), catalase (CAT) and also protein, malondialdehyde and ascorbic acid in vivo. Musa sapientum induced blood sugar reduction may be due to possible inhibition of free radicals and subsequent inhibition of tissue damage induced by alloxan. The antidiabetic activity observed in this plant may be attributed to the presence of flavonoids, alkaloids, steroid and glycoside principles. PMID- 16260407 TI - Experimental neurodegeneration in hippocampus and its phytoremidation. AB - Present study reports cytological and biochemical changes associated with stress induced neurodegeneration in hippocampal subregion of the brain in animals subjected to physical stressors such as immobilization or swimming stress for specific period of time. Studies also demonstrate neuroprotective activity of herbal extract in brain. PMID- 16260408 TI - Philosophy in Phytopharmacology:Ockham's Razor versus synergy. AB - Philosophy profoundly influences the sciences. In research models Ockham's Razor is often utilized as an operational principle in understanding the outcomes of modeling. However this principle of parsimony may delay the progress of understanding phytochemical matrices and the emergence of synergy that can arise from their interaction with human biology. Presented is a philosophical argument for synergy and medicinal plants as well as multiple examples of synergic systems occurring in the literature. PMID- 16260409 TI - Hypoglycemic and antiperglycemic effects of Semecarpus anacardium linn in normal and alloxan-induced diabetic rats. AB - The effect of ethanolic extract of dried nuts of Semecarpus anacardium on blood glucose level was investigated in both normal and alloxan induced diabetic rats. The blood glucose levels were measured at 0, 1, 2 and 3 hours after the treatment. The ethanolic extract of S. anacardium (100 mg/kg) reduced the blood glucose of normal rat from 84 +/- 1.4 to 67 +/- 1.7 mg/dl, 3 hours after oral administration of the extract (P < 0.05). It also significantly lowered blood glucose level in alloxan induced diabetic rat from 325 +/- 2.2 to 144 +/- 1.4 mg/dl, 3 hours after oral administration of the extract (P < 0.05). The antihyperglycemic activity of S. anacardium was compared with tolbutamide, an oral hypoglycemic agent. PMID- 16260410 TI - An evidence-based systemic review Echinacea E. angustifolia DC, E. pallida, E. purpurea by the Natural Standard Research Collaboration. PMID- 16260411 TI - Smoking status and mammography among women aged 50-75 in the 2002 behavioral risk factor surveillance system. AB - BACKGROUND: Since the late 1980's, data show an association between smoking status and mammography. Women smokers reported 12-15% lower rates than non smokers. This study investigated whether an association persists in a recent national-level database. METHODS: The sample was women aged 50-75 years from the 2002 Behavioral Risk Factor Surveillance System (N = 52,300). Analyses used two definitions of recent mammography, one based on a one-year interval between exams, the other on a two-year interval. Smoking was classified as current, former, or never. Other covariates included sociodemographic variables, health practices, insurance status and usual source of care. RESULTS: The rate of mammography for current smokers was 16% lower than for never smokers for the past year interval, and 14.0% lower for the two-year interval. Multiple logistic regression supported the smoking/mammography association. Other covariates associated with lower mammography on both dependent variables were recent Pap test, health insurance/usual source of care, recent dental visit, seat belt use, marital status and age. CONCLUSIONS: Reasons for the persistent association between smoking and mammography must be determined. These variables will inform interventions with women who smoke. A key question is whether to intervene on smoking directly or whether mediating variables are sufficient targets of intervention. PMID- 16260412 TI - The relationship between childhood adverse experiences and disability due to physical health problems in a community sample of women. AB - The goal of this study was to examine the association of physical and sexual abuse in childhood, poverty, parental substance abuse problems and parental psychiatric problems with disability due to physical health problems in a community sample of women. We included 4,243 women aged 15-64 years from the Ontario Mental Health Supplement in the analysis. The associations were tested by multiple logistic regression. Ten percent of women had a disability due to physical health problems. Among women with a disability, approximately 40% had been abused while growing up. After controlling for income and age, disability showed the strongest association with childhood physical abuse, parental education less than high school and parental psychiatric disorder. The association with child sexual abuse was not significant. Given the high correlation between abuse and disability due to physical health problems, it is important to investigate approaches to identify women who are at increased risk of subsequent impairment. PMID- 16260413 TI - The feminization of the physician assistant profession. AB - Although the physician assistant profession has historically been male-dominated, women now comprise over sixty percent of physician assistants (PAs) in the U.S. This paper explores the reason for the increase of women into the physician assistant profession in recent decades and whether gender differences exist in how PAs are utilized. Twenty-one qualitative interviews with male and female physician assistants and key informants were conducted to assess the reasons for the influx of women. In addition, data from the American Academy of Physician Assistants Census Survey (n = 16, 569) were analyzed to assess current gender differences in employment characteristics of PAs. In the interviews, female PAs reported entering the profession because it allowed them to practice within the medical model without having the high expense and demanding schedule of medical school. In fact, they claimed that the profession was quite compatible with family life. Significant gender differences were found in work characteristics, primary employer type, and practice specialty. Although women tend to concentrate in practice areas of women and children's health, evidence suggests that they are moving beyond these traditional roles into areas such as internal medicine and surgery. PMID- 16260414 TI - Serostatus disclosure to sexual partners by HIV-infected women before and after the advent of HAART. AB - HIV-positive individuals have been encouraged by public health officials to disclose their HIV status to sexual partners. In deciding what to do, however, they must weigh what they see as the potential costs and benefits of disclosing or not disclosing. In the present report we examine the reasons women offer for disclosing or not disclosing their serostatus and the reactions to that disclosure among two matched samples of HIV-infected women. The first sample was interviewed in 1994-1996, before the widespread availability of HAART; while the second sample was interviewed from 2000-2003 after these medications were widely in use. The findings reveal striking similarities between the two time periods in women's reasons for sharing or not sharing their status with partners, and the reactions to disclosure they experienced. The reconceptualization of AIDS as a chronic illness, rather than an acute fatal one, did not appear to have diminished women's felt responsibility to share their diagnosis with potential sexual partners nor their fear that disclosure would be met with rejection. The data revealed that disclosure remains a highly stressful event for HIV-infected women and that they experience considerable emotional suffering as a result of the diminished sense of self-worth and physical attractiveness brought about by their diagnosis. These findings suggest that intervention efforts toward reducing the stigmatization of HIV/AIDS and for assisting women to manage the stress of disclosure and non-disclosure and address women's feelings of self-worth continue to be needed despite the advent of HAART. PMID- 16260415 TI - Disordered eating, trauma, and sense of community: examining women in substance abuse recovery homes. AB - Women with substance-related disorders are likely to suffer from disordered eating and past traumatic experience, issues that might inhibit the recovery process. The present study determined the prevalence of co-morbidity of disordered eating, trauma, and substance-related disorders among 60 women living in one of 15 mutual help substance abuse recovery homes that establish new social networks for residents. Psychological sense of community was also examined to determine whether residents were able to obtain support that may have previously been absent. Results indicated that women with co-existing disordered eating and substance-related disorders, as well as women who have experienced trauma benefit from democratic, independent-living environments. Implications for treatment and future research are discussed. PMID- 16260416 TI - The role of the media in public participation: framing and leading. AB - This article focuses on the framing of women's health services in the context of restructuring dictated by health system regionalization. By examining the archives of a local newspaper and the minutes and documents of one of the key organizations involved in restructuring after regionalization, it was possible to examine the public discourse of the time and subsequently the journalists' and the readerships' understandings of women's health. The evidence suggests that the Salvation Army was instrumental in setting the tone that was taken by the media in framing the issues around the closure and move of its Grace Women's Health Centre. While the Calgary Health Region was successful in bringing the Grace under its mandate and organizational control, it was the Salvation Army, with its highly visible and powerful fundraising arm and its advocacy for holistic women's health that caught the public's attention. The internal discourse tracked some of the emerging issues, known only to those involved at managerial levels within the health system, but the public discourse kept women centered in decisions regarding the partnership. Women from many constituencies must continue to participate in the public policy realm to ensure that women's health remains an issue in health reform. PMID- 16260417 TI - Factors related to physical activity adherence in women: review and suggestions for future research. AB - Approximately 50 percent of individuals who start an exercise program withdraw within 6 months. Thus, many individuals withdraw before health benefits have been realized. This is a disconcerting statistic considering the well known benefits of physical activity for decreasing risk of hypokinetic diseases and improving quality of life. The literature has suggested a plethora of factors to increase the number of individuals who initiate a physical activity program. However, little is known about the factors that keep women exercising-otherwise known as exercise adherence. The purpose of this paper is to: (a) systematically review the quantitative literature to discern the major factors contributing to adherence to physical activity in women and men and make recommendations for specific gender-based considerations that are important when designing PA interventions for women, and (b) suggest areas of future research related to increasing adherence to physical activity in women. Key factors reviewed in this paper may be useful in developing efficacious physical activity programs for women. PMID- 16260418 TI - Regulation of KiSS-1 metastasis suppressor gene expression in breast cancer cells by direct interaction of transcription factors activator protein-2alpha and specificity protein-1. AB - KiSS-1 has been shown to function as a tumor metastasis suppressor gene and reduce the number of metastases in different cancers. The expression of KiSS-1 or KiSS1, like other tumor suppressor, is commonly reduced or completely ablated in a variety of cancers via an unknown mechanism. Here we show that the loss of KiSS 1 expression in highly metastatic breast cancer cell lines correlates directly with the expression levels of two transcription factors, activator protein-2alpha (AP-2alpha) and specificity protein 1 (Sp1), which synergistically activate the transcriptional regulation of KiSS-1 in breast cancer cells. Although the KiSS-1 promoter contains multiple AP-2alpha binding elements, AP-2alpha-mediated regulation occurs indirectly through Sp1 sites, as determined by deletion and mutation analysis. Overexpression of AP-2alpha into highly metastatic breast cell lines did not alter KiSS-1 promoter-driven luciferase gene activity. However, co transfection of AP-2alpha wild-type or the dominant negative form of AP-2 lacking its C-terminal DNA-binding domain, AP-2B, together with Sp1, increased KiSS-1 promoter activity dramatically, suggesting that AP-2alpha regulation of KiSS-1 transcription does not require direct binding to the KiSS-1 promoter. Furthermore, we demonstrated that AP-2alpha directly interacted with Sp1 to form transcription complexes at two tandem Sp1-binding sites of the promoter to activate KiSS-1 transcription. Together, our results indicate that AP-2alpha and Sp1 are strong transcriptional regulators of KiSS-1 and that loss or decreased expression of AP-2alpha in breast cancer may account for the loss of tumor metastasis suppressor KiSS-1 expression and thus increased cancer metastasis. PMID- 16260419 TI - Interferon-alpha-induced expression of phospholipid scramblase 1 through STAT1 requires the sequential activation of protein kinase Cdelta and JNK. AB - Phospholipid scramblase 1 (PLSCR1), a calcium-binding protein that either inserts into the plasma membrane or binds to genomic DNA in the nucleus, has been shown to contribute to the cell proliferation, differentiation, and apoptosis as well as antiviral activity of interferon (IFN). The expression of PLSCR1 protein is also known to be markedly increased in response to IFN and to some differentiation inducing agents such as all-trans retinoic acid, but the precise mechanisms of this response remain to be investigated. In this study, we show that the protein kinase Cdelta (PKCdelta)-specific inhibitor rottlerin and the dominant negative mutant of PKCdelta significantly antagonized IFN-induced PLSCR1 expression. The influence of PKCdelta on IFN-mediated induction of PLSCR1 was dependent upon the phosphorylation of STAT1 at Ser-727. Furthermore, PKCdelta mediated activation of STAT1 required the activation of JNK, as the inhibition of JNK activity by its specific inhibitor or transfection of its dominant negative mutant suppressed both serine phosphorylation of STAT1 and PLSCR1 expression but not the activation of PKCdelta. In conclusion, our results suggest that the induction of PLSCR1 transcription through STAT1 depends upon sequential activation of PKCdelta and JNK. PMID- 16260420 TI - Shift from Conn's syndrome to Cushing's syndrome in a recurrent adrenocortical carcinoma. AB - OBJECTIVE: Adrenocortical tumors may originate from the zona glomerulosa, zona fasciculata, or zona reticularis and be associated with syndromes due to overproduction of mineralocorticoids, glucocorticoids, or androgens respectively. We report an unusual case of recurrent adrenocortical carcinoma (ACC), which seems to contradict the paradigm of functional adrenal zonation. CASE REPORT: A male patient presented with severe primary aldosteronism due to an ACC, which relapsed after adrenalectomy and adjuvant mitotane therapy. After removal of the tumor recurrence and eight cycles of chemotherapy with etoposide, doxorubicin and cisplatin, the patient presented again with ACC masses, but in association with overt Cushing's syndrome and normal aldosterone levels. METHODS AND RESULTS: Extensive pathologic examination showed that this shift in steroid hormone production was paralleled by an attenuation of tumor cell atypia and polymorphism, whereas gene expression profile analysis demonstrated a change in expression of adrenal steroidogenic enzymes. Moreover, cancer progression was associated with overexpression of the inhibin-alpha subunit, which could have contributed to the phenotypic changes. CONCLUSIONS: This case of recurrent ACC demonstrates that adrenocortical cells can reverse their differentiation program during neoplastic progression and change their specific hormone synthesis, as a consequence of modifications in the expression profile of steroidogenic enzymes and cofactors. We hypothesize that this shift in steroid hormone secretion is a consequence of chromosome amplification induced by chemotherapy. These findings, besides opening new perspectives to study adrenocortical cell plasticity and potential, demonstrate how conventional clinical and pathologic evaluation can be combined with genomic analysis in order to dissect thoroughly the biology of cancer. PMID- 16260421 TI - High prevalence of suspicious cytology in thyroid nodules associated with positive thyroid autoantibodies. AB - OBJECTIVE: We assessed the association between thyroid autoimmunity and thyroid cancer in a retrospective series of unselected thyroid nodules submitted to fine needle aspiration cytology (FNAC) to avoid the selection bias of surgical series. SUBJECTS AND METHODS: Ultrasound (US)-guided FNACs were obtained from 590 unselected consecutive patients with single thyroid nodules and positive (ATA + , n = 197) or negative (ATA - , n = 393) serum anti-thyroid antibody (ATA). Cytological results were classified in three classes of increased risk of malignancy: low risk or benign (class II); indeterminate risk (class III); and suspect or malignant (class IV). RESULTS: A higher prevalence of class III (28.9% vs 21.4%, P < 0.05) and class IV (18.8% vs 9.2%, P < 0.001) and lower prevalence of class II (52.3% vs 69.5%, P < 0.001) were found in ATA + vs ATA - nodules respectively. By multivariate logistic regression analysis ATA + conferred a significant risk (odds ratio (OR): 2.29 (95% confidence interval (CI): 1.39 3.76)) for class IV cytology independently from age and sex. In 106 patients where thyroidectomy was carried out, thyroid cancer was found in 54/61 (88.5%) patients with class IV nodules (with similar positive predictive value for cancer in ATA + (96.4%) and ATA- (81.8%) nodules), in 6/31 (19.3%) of class III nodules (all ATA - ) and in none of 14 class II nodules. Non-specific cytological atypias from hyperplastic nodules in lymphocytic thyroiditis probably accounted for the different prevalence of cancer in class III ATA + and ATA - nodules. Histologically proven thyroid cancer (mostly papillary) was then observed in a higher proportion (27/197 = 13.7%) of ATA + , when compared with ATA - nodules (33/393 = 8.4%, P = 0.044), but the significance of this finding is limited by the low number of class II nodules operated on. CONCLUSIONS: The presence of ATA + confers an increased risk of suspicious or malignant cytology in unselected thyroid nodules. Since ATA + is not responsible for increased false-positive class IV FNAC, our study provides indirect evidence supporting a significant association between thyroid carcinoma and thyroid autoimmunity, although further studies with a different design are needed for a definitive histological proof. PMID- 16260422 TI - Thyroid hormone increases mannan-binding lectin levels. AB - BACKGROUND: Recent studies have indicated the existence of causal links between the endocrine and immune systems and cardiovascular disease. Mannan-binding lectin (MBL), a protein of the innate immune system, may constitute a connection between these fields. METHODS: To test whether thyroid hormone regulates MBL levels, we studied eight patients with Graves' hyperthyroidism before and after methimazole therapy, eight healthy subjects before and after short-term experimental hyperthyroidism, and eight hypothyroid patients with chronic auto immune thyroiditis before and after L-thyroxine substitution. RESULTS: In all hyperthyroid patients, MBL levels were increased--median (range), 1886 ng/ml (1478-7344) --before treatment and decreased to 954 ng/ml (312-3222) after treatment (P = 0.01, paired comparison: Wilcoxon's signed ranks test). The healthy subjects had MBL levels of 1081 ng/ml (312-1578). Administration of thyroid hormones to these persons induced mild hyperthyroidism and increased MBL levels significantly to 1714 ng/ml (356-2488) (P = 0.01). Two of the eight hypothyroid patients had undetectably low levels of MBL both before and after L thyroxine substitution. The other six hypothyroid patients had decreased levels of MBL of 145 ng/ml (20-457) compared with 979 ng/ml (214-1533) after L-thyroxine substitution (P = 0.03, paired comparison: Wilcoxon's signed ranks test). CONCLUSION: Our data show that thyroid hormone increases levels of MBL. MBL is part of the inflammatory complement system, and this modulation of complement activation may play a role in the pathogenesis of a number of key components of thyroid diseases. PMID- 16260423 TI - Post-surgical use of radioiodine (131I) in patients with papillary and follicular thyroid cancer and the issue of remnant ablation: a consensus report. AB - OBJECTIVE: To determine, based on published literature and expert clinical experience, current indications for the post-surgical administration of a large radioiodine activity in patients with differentiated thyroid cancer. DESIGN AND METHODS: A literature review was performed and was then analyzed and discussed by a panel of experts from 13 European countries. RESULTS: There is general agreement that patients with unifocal microcarcinomas = 1 cm in diameter and no node or distant metastases have a <2% recurrence rate after surgery alone, and that post-surgical radioiodine confers recurrence and cause-specific survival benefits in patients, strongly suspected of having persistent disease or known to have tumor in the neck or distant sites. In other patients, there is limited evidence that after complete thyroidectomy and adequate lymph node dissection performed by an expert surgeon, post-surgical radioiodine provides clear benefit. When there is any uncertainty about the completeness of surgery, evidence suggests that radioiodine can reduce recurrences and possibly mortality. CONCLUSION: This survey confirms that post-surgical radioiodine should be used selectively. The modality is definitely indicated in patients with distant metastases, incomplete tumor resection, or complete tumor resection but high risk of recurrence and mortality. Probable indications include patients with tumors >1 cm and with suboptimal surgery (less than total thyroidectomy or no lymph node dissection), with age <16 years, or with unfavorable histology. PMID- 16260424 TI - Interdependence of lean body mass and total body water, but not quality of life measures, during low dose GH replacement in GH-deficient adults. AB - Lean body mass (LBM) and total body water (TBW) are reduced in GH-deficient (GHD) adults and alter with GH replacement. Whether these parameters are interdependent and whether alterations in their homeostasis contribute to the perceived quality of life (QOL) deficit in GHD remains unclear. In this study, IGF-I, body composition by whole-body dual-energy X-ray absorptiometry, TBW by deuterium dilution (D(2)O) and two validated QOL instruments - psychological general well being schedule (PGWB, generic, 6 domains; lower score worse QOL) and assessment of GH deficiency in adults (AGHDA, disease orientated; higher score worse QOL) were studied at baseline and after 3 and 6 months of GH replacement in thirty GHD adults. Patients with diabetes insipidus, and cardiac and renal failure were excluded. Median age-adjusted IGF-I standard deviation score increased from -3.40 (-6.40 to -1.60) to -0.2 (-1.88 to 0.78) (P < 0.0001) at a median daily GH dose of 0.4 mg. During treatment, LBM increased from 47.4 +/- 10.7 kg at baseline to 49.5 +/- 10.8 kg at 6 months (P = 0.0008), and fat mass decreased from 28.0 +/- 12.1 kg at baseline to 27.2 +/- 12.6 kg at 6 months (P = 0.0004). A non significant trend towards an increase in TBW was observed (mean 1.7 kg, P = 0.08). The PGWB score increased from 62.9 +/- 20.6 to 73.7 +/- 21.7 (P = 0.0006). The AGHDA score decreased from 13.7 +/- 7.3 to 8.75 +/- 7.75 (P = 0.0002). At each time point, a linear correlation between LBM and TBW was demonstrated, defined by TBW = (0.972 x LBM)-10.6. However, only a weakly positive correlation existed between the percentage changes in these variables (R = 0.40, P = 0.04). No correlations were demonstrated between QOL measures and body composition. The change in LBM with physiological GH replacement correlates weakly with change in TBW, therefore factors other than TBW may also contribute to the LBM changes. Improved QOL with GH replacement is not explained by favourable changes in body composition. PMID- 16260425 TI - Administration of recombinant human GHRH-1,44-amide for 3 months reduces abdominal visceral fat mass and increases physical performance measures in postmenopausal women. AB - OBJECTIVE: A recent study indicated that twice-daily s.c. administration of a high dose of recombinant human GHRH-1,44-amide (GHRH) for 90 days can alter body composition in healthy older men. No data establish whether this is also true in postmenopausal women. The present study tests the hypothesis that the same GHRH regimen applied in women will: (i) elevate both IGF-I and GH concentrations; and (ii) reduce abdominal visceral fat mass, augment total body water and enhance functional performance. DESIGN: Ten postmenopausal volunteers underwent baseline study and then received 1 mg GHRH twice daily s.c. for 3 months. METHODS: Statistical comparisons were made with pre-intervention baseline data. RESULTS: GHRH administration stimulated: (i) a mean 98 +/- 14% elevation of overnight GH concentrations after administration of the peptide for 1 and 3 months (P < 0.005); (ii) a sustained 71 +/- 3.5% rise in IGF-I concentrations over the interval from 2 weeks to 3 months (P < 0.0012); (iii) a 16 +/- 7% reduction in abdominal visceral fat mass (P = 0.029) and a 14 +/- 5% increase in tri-tiated water space (P < 0.025); (iv) an abbreviation of the times required to walk 30 m (P = 0.015) and ascend two flights of stairs (P = 0.003). Most (70%) subjects experienced local skin reactivity. There were no systemic adverse events. CONCLUSIONS: A 3-month regimen of GHRH supplementation in postmenopausal women can stimulate GH and IGF-I production, reduce abdominal visceral fat and improve selected measures of physical performance, while inducing significant local skin reactivity. PMID- 16260426 TI - Efficacy and safety of Monascus purpureus Went rice in subjects with hyperlipidemia. AB - OBJECTIVE: The purpose of this study was to assess the lipid-lowering effect of Monascus purpureus Went rice on serum lipids in patients with hyperlipidemia, and to assess its safety by reporting adverse events and clinical laboratory measurements. DESIGN AND METHODS: This was a randomized, double-blind, placebo controlled study. In all, 79 patients (aged 23-65 years) with a mean baseline low density lipoprotein cholesterol (LDL-C) level of 5.28 mmol/l (203.9 mg/dl) received a twice daily dose of placebo or Monascus purpureus Went rice (600 mg) for 8 weeks. RESULTS: At week 8, Monascus purpureus Went rice therapy reduced LDL C by 27.7%, total cholesterol by 21.5%, triglycerides by 15.8% and apolipoprotein B by 26.0%. High-density lipoprotein cholesterol and apolipoprotein A-I levels were increased by 0.9 and 3.4% respectively (not significant). No patient in the Monascus purpureus Went rice treatment group had an alanine aminotransferase (ALT), aspartate aminotransferase (AST) or creatine phosphokinase (CPK) measurement that was > or = 3 times the upper limit of normal at week 4 and week 8. CONCLUSION: Monascus purpureus Went rice significantly reduced LDL-C, total cholesterol, triglycerides and apolipoprotein B levels, and was well tolerated in patients with hyperlipidemia. However, this study only provides data from an 8 week trial and long-term safety and efficacy data are needed. PMID- 16260427 TI - The tumour necrosis factor (TNF)-alpha system is activated in accordance with pulse pressure in normotensive subjects with type 1 diabetes mellitus. AB - OBJECTIVE: Pulse pressure (PP) and inflammation are important predictors of cardiovascular disease (CVD), even in the normotensive. The age-related increase in PP can be diagnosed up to 20 years earlier in subjects with type 1 diabetes mellitus (T1DM) than in the general population. Some evidence suggests that PP can stimulate inflammation. Our aim was to study the relationship between PP and plasma inflammatory proteins in normotensive subjects with T1DM. DESIGN: This was a cross-sectional study of a group of normotensive (<140/80 mmHg) subjects diagnosed with T1DM 14 years before. None of them had clinically proven CVD or inflammatory conditions or were on antiplatelet, antihypertensive, anti inflammatory or lipid-lowering treatment. METHODS: The following information was recorded: sex, age, body-mass index (BMI), waist-to-hip ratio (WHR), systolic blood pressure (SBP), diastolic blood pressure (DBP), PP, mean blood pressure (MBP), smoking, alcohol intake, insulin dose, lipid profile, HbA1c, microvascular complications, and plasma concentrations of soluble receptor types 1 and 2 of tumour necrosis factor (TNF)-alpha (sTNFR1 and sTNFR2, respectively), interleukin 6, C-reactive protein, adiponectin and leptin. RESULTS: A total of 112 subjects were evaluated (aged 27.4+/-6.6 years, 52.7% women, BMI: 20.4+/-2.7 kg/m2, WHR: 0.82+/-0.09, SBP: 112+/-12 mmHg, DBP: 68+/-9 mmHg, PP: 45+/-9 mmHg, MBP: 82+/-9 mmHg, HbA1c: 8.2% (7.3-9.0%), 41.1% microvascular complications). After adjusting for potential confounders, only inflammatory markers of the TNF-alpha system correlated significantly with PP (Pearson correlation coefficient between sTNFR1 and PP: r = 0.215, P = 0.030; and between PP and sTNFR2: r = 0.238, P = 0.020). CONCLUSION: In normotensive subjects with T1DM after 14 years of diagnosis, the activation of the TNF-alpha system is positively associated with PP levels. This finding might suggest a pathogenic role of the TNF-alpha system in the development of cardiovascular disease in T1DM. PMID- 16260428 TI - The expression of wild-type pendrin (SLC26A4) in human embryonic kidney (HEK 293 Phoenix) cells leads to the activation of cationic currents. AB - OBJECTIVE: The SLC26A4 protein (pendrin) seems to be involved in the exchange of chloride with other anions, therefore being responsible for iodide organification in the thyroid gland and the conditioning of the endolymphatic fluid in the inner ear. Malfunction of SLC26A4 leads to Pendred syndrome, characterized by mild thyroid dysfunction often associated with goiter and/or prelingual deafness. The precise function of the SLC26A4 protein, however, is still elusive. An open question is still whether the SLC26A4-induced ion exchange mechanism is electrogenic or electroneutral. Recently, it has been shown that human pendrin expressed in monkey cells leads to chloride currents. METHODS: We overexpressed the human SLC26A4 isoform in HEK293 Phoenix cells and measured cationic and anionic currents by the patch-clamp technique in whole cell configuration. RESULTS: Here we show that human pendrin expressed in human cells does not lead to the activation of chloride currents, but, in contrast, leads to an increase of cationic currents. CONCLUSION: Our experiments suggest that the SLC26A4-induced chloride transport is electroneutral when expressed in human cellular systems. PMID- 16260429 TI - Expression of vascular endothelial growth factor (VEGF) and its receptors in thyroid carcinomas of follicular origin: a potential autocrine loop. AB - OBJECTIVE: The aim of this study was to clarify the role of vascular endothelial growth factor (VEGF) and VEGF receptor (VEGFR) pathways in thyroid tumourigenesis. METHODS: We examined VEGF, VEGFR-1 and VEGFR-2 expression on 34 papillary thyroid carcinomas (PTCs), 18 follicular thyroid carcinomas (FTCs), eight poorly differentiated thyroid carcinomas (PDTCs) and on a thyroid tumour derived cell line (NPA'87) by immunohistochemistry, reverse transcriptase PCR, immunofluorescence and Western blotting. RESULTS: We have demonstrated that VEGF expression was significantly (P < 0.05) more prevalent in PTCs (79%) than in FTCs (50%) or PDTCs (37%). Similarly, 76% of PTCs, 83% of FTCs and 25% of PDTCs expressed VEGFR-1, whereas 68% of PTCs, 56% of FTCs and 37% of PDTCs expressed VEGFR-2. Coexpression of VEGF and its receptors was observed in 50% of PTCs, 39% of FTCs and 12% of PDTCs, raising the possibility that VEGF may signal in an autocrine loop in these neoplasias, as observed previously for other types of cancer. In agreement with the idea that autocrine VEGF signalling plays an important role in thyroid carcinogenesis, the blockade of either VEGF or its receptors with neutralizing antibodies significantly reduced cell viability and increased apoptosis levels of the VEGFR-positive thyroid tumour cell line NPA'87. CONCLUSIONS: Our results highlight a previously undefined VEGF autocrine action in thyroid carcinomas which could play a crucial role in tumour cell survival and could represent a useful therapeutic target for thyroid tumours. PMID- 16260431 TI - Increased frequency of subclinical hypothyroidism and thyroid-associated antibodies in siblings of children and adolescents with type 1 diabetes mellitus. PMID- 16260430 TI - Genetic influence of an ACTH receptor promoter polymorphism on adrenal androgen secretion. AB - OBJECTIVE: Adrenocorticotropic hormone (ACTH) is the primary secretagogue stimulating secretion of adrenal androgens (AA). Yet, genetic and environmental factors are assumed to play a determining role in the regulation of their biosynthesis and thus might explain the high variability of AA levels. Here we investigate the influence of an ACTH receptor promoter polymorphism affecting ACTH receptor gene transcription on ACTH-dependent dehydroepiandrosterone (DHEA) secretion. DESIGN: We recently reported a polymorphism within the transcription initiation site of the ACTH receptor gene promoter that alters the consensus sequence from CTC to CCC at -2 bp. This results in lower promoter activity in vitro and is associated with impaired cortisol response to ACTH stimulation in vivo. We now studied 14 normal, lean volunteers aged 20-35 years (eight CTC/CTC and six CCC/CCC carriers) in a 6-h ACTH stimulation test. METHODS: After overnight dexamethasone suppression, ACTH1-24 was administered continuously in each subject with hourly increasing doses (120-3840 ng/m2 body surface area/h) within a 6-h period. On a separate day, baseline DHEA samples were collected. RESULTS: In the 6-h ACTH stimulation test, CTC/CTC carriers showed a significantly higher DHEA response than CCC/CCC carriers (area under the curve: 19 367 +/- 2919 vs 11 098 +/- 1241 nmol/l per min; P < 0.04, Mann-Whitney U test). In contrast, baseline DHEA concentrations did not differ between groups. CONCLUSION: These data demonstrate that genetic variations within the ACTH receptor promoter result in decreased DHEA secretion. Thus, we might have identified one of the genetic factors responsible for variation in ACTH-dependent DHEA secretion. PMID- 16260434 TI - Feasibility and preliminary outcomes from a pilot study of a brief psychological intervention for families of children newly diagnosed with cancer. AB - OBJECTIVE: To report initial feasibility and outcome from a pilot study of a new three-session intervention for caregivers of children newly diagnosed with cancer, Surviving Cancer Competently Intervention Program-Newly Diagnosed (SCCIP ND). METHOD: Nineteen families (38 caregivers) were randomly assigned to SCCIP-ND or treatment as usual subsequent to learning of their child's illness. The study design included pre- and 2-month postintervention assessments, with state anxiety and posttraumatic stress symptoms as outcomes. Feasibility was based on therapist feedback and supervision, program evaluations, and data from study-tracking procedures. RESULTS: SCCIP-ND appears to be an acceptable intervention that can be used successfully with caregivers over the first few months after diagnosis. Recruitment and retention data document feasibility but also highlight challenges. Preliminary outcome data show changes in the desired direction [e.g., reduced anxiety and parental posttraumatic stress symptoms (PTSS)]. CONCLUSIONS: The pilot data are supportive of the value and challenges of developing evidence based family interventions in pediatric psychology. PMID- 16260435 TI - Multisystemic treatment of poorly controlled type 1 diabetes: effects on medical resource utilization. AB - OBJECTIVE: To determine whether multisystemic therapy (MST), an intensive, home based psychotherapy, could decrease rates of hospital utilization and related costs of care among adolescents with poorly controlled type diabetes. METHODS: Thirty-one adolescents were randomly assigned to receive either MST or standard care. MST lasted approximately 6 months, and all participants were followed for 9 months. Rates of inpatient admissions and emergency room (ER) visits were calculated for a 9-month prestudy period and during the 9 months of study participation. The relationship between changes in inpatient admissions and changes in metabolic control was also investigated. RESULTS: Intervention participants had a decreasing number of inpatient admissions from the baseline period to the end of the study, whereas the number of inpatient admissions increased for controls. Use of the emergency room did not differ. Related medical charges and direct care costs were significantly lower for adolescents receiving MST. Correlational analyses conducted with a subset of participants indicated that decreases in inpatient admissions were associated with improved metabolic control for MST but not control participants. CONCLUSIONS: Findings suggest that MST has the potential to decrease inpatient admissions among adolescents with poorly controlled type 1 diabetes. PMID- 16260436 TI - Family-based intervention to enhance infant-parent relationships in the neonatal intensive care unit. AB - OBJECTIVE: To examine how family-based interventions in the neonatal intensive care unit (NICU) may change parental knowledge and behaviors and decrease stress. METHODS: Eighty-four high-risk mother-infant dyads were randomly assigned to two intervention and one control groups. Group 1 (n = 28) participated in a demonstration of infant reflexes, attention, motor skills, and sleep-wake states. Group 2 (n = 31) viewed educational materials. Group 3 (n = 25), controls, participated in an informal discussion. Parent-infant interactions (Nursing Child Assessment Feeding Scale) were videotaped. Mothers completed measures of stress (Parenting Stress Index) and knowledge of infant cues (Knowledge of Preterm Infant Behavior Scale). RESULTS: Mothers in both intervention groups evidenced greater knowledge and more contingent and sensitive interactions with their infants than did the control group. Stress also differed across groups, and all mothers reported scores above norms. CONCLUSIONS: In a high-risk sample, short term, family-based NICU interventions may enhance mothers' knowledge, sensitivity, contingency, and stress. PMID- 16260437 TI - Brief report: family-based group intervention for young siblings of children with chronic illness and developmental disability. AB - OBJECTIVE: To evaluate the impact of a family-based group intervention for young siblings of children with chronic illness and developmental disability (CI/DD). METHODS: Forty-three healthy siblings (ages 4-7 years) of children with CI/DD and their parents participated in an intervention designed to address sibling challenges that cut across types of diagnostic conditions. The intervention consisted of six sessions of collateral and integrated sibling-parent groups. Measures of sibling knowledge, sibling sense of connectedness with other children in similar family circumstances, and sibling global functioning were collected before and after intervention. A subsample of 17 families completed a 3-month follow-up. RESULTS: Siblings' knowledge of the child's disorder and sibling connectedness increased significantly from pre- to posttreatment for both boys and girls, regardless of the nature of the brother or sister's condition. Sibling perceptions of self-competence increased from pre- to posttreatment, whereas parent reports of sibling behavioral functioning remained within the normal range. Improvements in sibling knowledge and connectedness maintained at follow up. Parent satisfaction with the program was high. CONCLUSIONS: Results support more controlled evaluations of family-based intervention to improve young sibling adaptation to CI/DD. PMID- 16260438 TI - Brief report: In-home family therapy for adolescents with poorly controlled diabetes: failure to maintain benefits at 6-month follow-up. AB - OBJECTIVE: To examine 6-month follow-up data on the effectiveness of in-home Behavioral Family Systems Therapy (BFST) for adolescents with poorly controlled diabetes, using a pilot and feasibility study. METHODS: Eighteen adolescents with poorly controlled diabetes received ten 90-min sessions of in-home BFST. Diabetes related functioning, general family functioning, and health status were assessed at baseline, immediately following treatment and 6-months after the treatment. RESULTS: Although the initial posttreatment follow-up evaluation indicated decreases in general family conflict, diabetes-related family conflict, and behavior problems, evaluation at a 6-month follow-up (N = 17) demonstrated that initial posttreatment improvements were no longer present for any of the variables assessed. Metabolic control remained unchanged from baseline to initial posttreatment as well as at 6-month follow-up. CONCLUSIONS: A plausible explanation for this finding is that participating families were experiencing distress that required longer-term treatment for enduring results, beyond what was employed in this study. Further research is necessary before in-home BFST can be considered an effective psychosocial intervention for adolescents with poorly controlled diabetes. PMID- 16260439 TI - Commentary: Involving families in psychological interventions in pediatric psychology: critical needs and dilemmas. PMID- 16260440 TI - Nitrogen reserves, spring regrowth and winter survival of field-grown alfalfa (Medicago sativa) defoliated in the autumn. AB - AIMS: The objective of the study was to characterize variations in proline, arginine, histidine, vegetative storage proteins, and cold-inducible gene expression in overwintering roots of field-grown alfalfa, in response to autumn defoliation, and in relation to spring regrowth and winter survival. METHODS: Field trials, established in 1996 in eastern Canada, consisted of two alfalfa cultivars ('AC Caribou' and 'WL 225') defoliated in 1997 and 1998 either only twice during the summer or three times with the third defoliation taken 400, 500 or 600 growing degree days (basis 5 degrees C) after the second summer defoliation. KEY RESULTS: The root accumulation of proline, arginine, histidine and soluble proteins of 32, 19 and 15 kDa, characterized as alfalfa vegetative storage proteins, was reduced the following spring by an early autumn defoliation at 400 or 500 growing degree days in both cultivars; the 600-growing-degree-days defoliation treatment had less or no effect. Transcript levels of the cold inducible gene msaCIA, encoding a glycine-rich protein, were markedly reduced by autumn defoliation in 'WL 225', but remained unaffected in the more winter-hardy cultivar 'AC Caribou'. The expression of another cold-inducible gene, the dehydrin homologue msaCIG, was not consistently affected by autumn defoliation. Principal component analyses, including components of root organic reserves at the onset of winter, along with yield and plant density in the following spring, revealed that (a) amino acids and soluble proteins are positively related to the vigour of spring regrowth but poorly related to winter survival and (b) winter survival, as indicated by plant density in the spring, is associated with higher concentrations of cryoprotective sugars in alfalfa roots the previous autumn. CONCLUSIONS: An untimely autumn defoliation of alfalfa reduces root accumulation of specific N reserves such as proline, arginine, histidine and vegetative storage proteins that are positively related to the vigour of spring regrowth but poorly related to winter survival. PMID- 16260441 TI - Co-ordinated growth between aerial and root systems in young apple plants issued from in vitro culture. AB - BACKGROUND AND AIMS: In several species exhibiting a rhythmic aerial growth, the existence of an alternation between root and shoot growth has been demonstrated. The present study aims to investigate the respective involvement of the emergence of new organs and their elongation in relation to this phenomenon and its possible genotypic variation in young apple plants. METHODS: Two apple varieties, X6407 (recently named 'Ariane') and X3305 ('Chantecler' x 'Baujade'), were compared. Five plants per variety, issued from in vitro culture, were observed in minirhizotrons over 4 months. For each plant, root emergence and growth were observed twice per week. Growth rates were calculated for all roots with more than two segments and the branching density was calculated on primary roots. On the aerial part, the number of leaves, leaf area and total shoot length were observed weekly. KEY RESULTS: No significant difference was observed between varieties in any of the final characteristics of aerial growth. Increase in leaf area and shoot length exhibited a 3-week rhythm in X3305 while a weaker signal was observed in Ariane. The primary root growth rate was homogeneous between the plants and likewise between the varieties, while their branching density differed significantly. Secondary roots emerged rhythmically, with a 3-week and a 2-week rhythm, respectively, in X3305 and 'Ariane'. Despite a high intra-variety variability, significant differences were observed between varieties in the secondary root life span and mean length. A synchronism between leaf emergence and primary root growth was highlighted in both varieties, while an opposition phase was observed between leaf area increments and secondary root emergence in X3305 only. CONCLUSION: A biological model of dynamics that summarizes the interactions between processes and includes the assumption of a feedback effect of lateral root emergence on leaf emergence is proposed. PMID- 16260444 TI - Self-rated health in relation to employment status during periods of high and of low levels of unemployment. AB - BACKGROUND: There is a need for more research on the health impact of changes in the national unemployment rate. Therefore, the present study was carried out to compare levels of self-rated health during periods of high and low levels of unemployment. METHODS: Data included cross-sectional interviews from the Swedish Survey of Living Conditions, which were based on random samples of inhabitants between 16 and 64 years of age living in Sweden. Data were collected for the period 1983-89, when unemployment levels were low (n = 35 562; 2.5%) and for the period 1992-97 when unemployment was high (n = 24 019; 7.1%). RESULTS: After adjusting for sociodemographic variables as well as long-term disease or handicap, the differences in self-rated health between the unemployed and employed were larger when unemployment levels were high in the 1990s, than when they were low in the 1980s. More groups of the unemployed were afflicted with poor health when unemployment was high, compared with when it was low. In 1992 97, being married, living in larger cities, or not having a long-term disease or handicap no longer buffered the negative effects on health among the unemployed. CONCLUSIONS: Poorer self-rated health among the unemployed seems to be an increasing public health problem during high levels of unemployment. PMID- 16260442 TI - Epigenetics and its implications for plant biology 2. The 'epigenetic epiphany': epigenetics, evolution and beyond. AB - SCOPE: In the second part of a two-part review, the ubiquity and universality of epigenetic systems is emphasized, and attention is drawn to the key roles they play, ranging from transducing environmental signals to altering gene expression, genomic architecture and defence. KEY ISSUES: The importance of transience versus heritability in epigenetic marks is examined, as are the potential for stable epigenetic marks to contribute to plant evolution, and the mechanisms generating novel epigenetic variation, such as stress and interspecific hybridization. FUTURE PROSPECTS: It is suggested that the ramifications of epigenetics in plant biology are immense, yet unappreciated. In contrast to the ease with which the DNA sequence can be studied, studying the complex patterns inherent in epigenetics poses many problems. Greater knowledge of patterns of epigenetic variation may be informative in taxonomy and systematics, as well as population biology and conservation. PMID- 16260446 TI - High prevalence of the plasmid-mediated quinolone resistance determinant qnrA in multidrug-resistant Enterobacteriaceae from blood cultures in Liverpool, UK. AB - OBJECTIVES: To determine the prevalence of the plasmid-mediated quinolone resistance qnrA gene in a selected collection of blood culture isolates of Enterobacteriaceae resistant to both ciprofloxacin and cefotaxime. METHODS: Over a 29 month period, a total of 47 non-repetitive isolates of Enterobacteriaceae resistant to both ciprofloxacin and cefotaxime were identified. Isolates were screened for the presence of the qnrA gene, class I integrons and bla(ESBL) by PCR. Transferability was examined by conjugation with the sodium azide-resistant Escherichia coli J53. All qnrA-positive isolates were examined for DNA relatedness by PFGE. RESULTS: A total of 15 of the 47 test isolates (32%) were positive for the qnrA gene, and included single isolates of E. coli and Citrobacter freundii, 4 Klebsiella pneumoniae and 9 Enterobacter cloacae. All 15 qnrA-positive isolates carried class 1 integrons, and 11 the extended-spectrum beta-lactamase gene bla(SHV-12). By PFGE two K. pneumoniae and three E. cloacae, respectively, were considered clonally but not temporally related. Plasmid transfer of quinolone resistance was only achieved with single isolates of K. pneumoniae and E. cloacae. Both plasmids carried class 1 integrons with a pSAL-1 like gene cassette arrangement intl1-aadA2-qacEDelta-sul1. CONCLUSIONS: In this selected group of ciprofloxacin- and cefotaxime-resistant bacteria, carriage of the qnrA gene was high (32%). This compares with <2.0% as demonstrated in worldwide studies of laboratory collections of ciprofloxacin-resistant bacteria. The majority of qnrA-positive isolates in our study originated from high dependency care units within our hospital, but were shown not to be clonal by PFGE. This is the first report of qnrA-positive Enterobacteriaceae in the United Kingdom. PMID- 16260445 TI - In vitro activity of pazufloxacin, tosufloxacin and other quinolones against Legionella species. AB - OBJECTIVES: The activities of pazufloxacin and tosufloxacin against Legionella spp. were evaluated in vitro and compared with those of other quinolones, macrolides and azithromycin. METHODS: The conventional MICs were determined by the microbroth dilution method. Intracellular activities of drugs were evaluated by a cfu count. The minimal extracellular concentration inhibiting intracellular growth of bacteria (MIEC) was determined by a colorimetric cytopathic assay. RESULTS: MICs of pazuloxacin and tosufloxacin at which 90% (MIC90) of isolates are inhibited in 76 different Legionella spp. strains (38 ATCC strains and 38 clinical isolates) were 0.032 and 0.016 mg/L, whereas the MIC90s of levofloxacin, ciprofloxacin, garenoxacin, erythromycin, clarithromycin and azithromycin were 0.032, 0.032, 0.032, 2.0, 0.125 and 2.0 mg/L, respectively. Pazufloxacin and tosufloxacin at 4x MIC inhibited intracellular growth of Legionella pneumophila SG1 (80-045 strain), as did other quinolones, clarithromycin and azithromycin, whereas erythromycin at 4x MIC did not. MIECs of pazufloxacin, tosufloxacin, levofloxacin, ciprofloxacin and garenoxacin for the strain were 0.063, 0.004, 0.016, 0.032 and 0.008 mg/L respectively, which were superior to those of macrolides and azithromycin. Pazufloxacin showed potent activity against three additional clinical isolates of L. pneumophila SG1, one clinical isolate each of L. pneumophila SG3 and SG5, as well as Legionella micdadei, Legionella dumoffii and Legionella longbeachae SG1. CONCLUSIONS: Pazufloxacin and tosufloxacin, as well as other quinolones, were more potent than macrolides and an azalide. Present data warrant further study on the efficacy of these drugs in the treatment of Legionella infections. PMID- 16260447 TI - Responses to cardiovascular drugs during alcohol withdrawal. AB - AIM: To present findings on the kinetics and dynamics of cardiovascular drugs during alcohol withdrawal (AW), compared with that observed in remission. METHOD: Studies were reviewed and summarized. RESULTS: A single-dose study in alcoholic patients with propranolol, a beta-adrenergic antagonist, showed that the negative inotropic effect was decreased and the bradycardiac effect increased during AW as compared with during early remission. The hypotensive effect of isosorbid dinitrate, commonly used as a vasodilatator, was weaker at the onset of AW, being associated with the decreased bioavailability of the drug. Verapamil, which is a L-type Ca2+ channel antagonist, produced a bradycardiac effect at the onset of AW, but no effect was observed in early remission. The effect was probably due to changes in L-type Ca2+ channels because no differences in verapamil concentrations between AW and early remission were observed. CONCLUSION: Taken together, AW modifies the dynamics and kinetics of cardiac drugs, which may have an impact on the treatment of alcoholic patients with cardiac diseases. PMID- 16260448 TI - Alcohol hangover effects on measures of affect the morning after a normal night's drinking. AB - AIM: To investigate the effects of students' usual levels of alcohol consumption on aspects of mood and anxiety the following morning. METHODS: Students were recruited who consumed their usual quantity of any type of alcoholic beverage in their chosen company and then completed assessments of the effects the following day. The timing of drinking was restricted to the period between 22:00 and 02:00 h the night before testing as these are the most popular hours for consuming alcohol in the population under investigation. The testing included an assessment of mood and anxiety; testing was also performed after an evening of abstinence (no hangover condition), following a counterbalanced repeated measure design, with time of testing and order of testing as 'between participant' factors. Forty eight student social drinkers (33 women, 15 men) aged between 18 and 43 years were tested, with a 1 week interval between test sessions. RESULTS: Males reported consuming on average 14.7 units and females 10.5 units the night before testing. On the morning after alcohol consumption, ratings of alertness and tranquility were lower than the ratings the morning following an evening of abstinence at both 11:00 and 13:00 h and the post intoxication physical symptoms, emotional symptoms and symptoms of fatigue persisted throughout the morning. CONCLUSION: Heavy alcohol consumption lowers mood, disrupts sleep, increases anxiety and produces physical symptoms, emotional symptoms and symptoms of fatigue throughout the next morning. PMID- 16260449 TI - The association of fat patterning with blood pressure in rural South African children: the Ellisras Longitudinal Growth and Health Study. AB - BACKGROUND: Hypertension is one of the major causes of death in developed and underdeveloped nations. Essential hypertension and obesity may have their inception in childhood, with little data in African children to support these findings. Objectives were to determine the prevalence of overweight and hypertension in rural children in South Africa. Additionally, the association between fat-patterning ratios and blood pressure (BP) was investigated. METHODS: Data were collected from 1884 subjects (967 boys and 917 girls), aged 6-13 years, participating in the Ellisras Longitudinal Study. Height; weight; and triceps, biceps, subscapular, and suprailiac skinfolds were measured according to the protocol of the International Society for the Advancement of Kinanthropometry. Skinfold ratio was used as an indicator of the central pattern of body fat. Internationally recommended cut-off points for body mass index (BMI) were used. Hypertension, defined as the average of three separate BP readings where the systolic BP or diastolic BP is >or=95th percentile for age and sex, was determined. RESULTS: The prevalence of hypertension ranged from 1 to 5.8% for boys and 3.1 to 11.4% for girls, and that of overweight from 1.1 to 2.9% for boys and 0.6 to 4.6% for girls. The association between high systolic BP and high BMI was -3.0, while that for high diastolic BP and high BMI was -0.68. CONCLUSIONS: The prevalence of hypertension is evident from the age 6 years for girls, while that of overweight was low. Overweight became evident from the age 10 to 13 years for both sexes. A significant association between high diastolic BP and high BMI was noted, while children with low BMIs were less likely to be hypertensive. Investigating habitual physical activity, fitness and dietary patterns will shed more light on the association of fat patterning and BP in this population. PMID- 16260450 TI - Maternal smoking during pregnancy in relation to child overweight: follow-up to age 8 years. AB - BACKGROUND: Data from several studies indicate that children of mothers who smoked during pregnancy may be at a risk of overweight compared with children of non-smoking mothers. The size of this relation, however, is unclear, as is the age at which it becomes detectable. METHODS: Prospective data for 34 866 children enrolled in the US Collaborative Perinatal Project were analysed to examine maternal pregnancy smoking in relation to weight, height, and body mass index (BMI) in offspring at ages 1, 3, 4, 7, and 8 years. RESULTS: Compared with offspring of non-smokers, children of smokers had (i) weight that was lower at birth but then quickly equalled or exceeded that of non-smokers, (ii) consistently decreased height, and (iii) increased risk of overweight, particularly in girls. For example, at age 7 years, the adjusted odds ratio of BMI>or=85th percentile in boys of mothers who smoked on an average>or=20 cigarettes per day while pregnant was 1.22 (95% confidence interval 1.03-1.46), and in girls it was 1.30 (1.08-1.56). CONCLUSIONS: In these data, maternal smoking during pregnancy was associated with a modest increase in risk of overweight in children before the age of 8 years. PMID- 16260451 TI - Longevity of popes and artists between the 13th and the 19th century. PMID- 16260452 TI - Commentary: Donald Budd Armstrong (1886-1968)--pioneering tuberculosis prevention in general practice. PMID- 16260453 TI - Commentary: the first Framingham Study--a pioneer in community-based participatory research. PMID- 16260456 TI - Improving quality of care for depression: the German Action Programme for the implementation of evidence-based guidelines. AB - ISSUE: Depressive disorders are of great medical and political significance. The potential inherent in achieving better guideline orientation and a better collaboration between different types of care is clear. Throughout the 1990s, educational initiatives were started for implementing guidelines. Evidence-based guidelines on depression have been formulated in many countries. PURPOSE: This article presents an action programme for structural, educational, and research related measures to implement evidence-based care of depressive disorders in the German health system. The starting points of the programme are the 'Guidelines Critical Appraisal Reports' of the 'Guideline Clearing House' and measures from the 'Competence Network on Depression and Suicidality' (CNDS) funded by the Federal Ministry of Education and Research. The article gives an overview of the steps achieved as recommended by the Guidelines Critical Appraisal Reports and the ongoing transfer process into the German health care system. RESULTS: The action programme shows that comprehensive interventions to develop and introduce evidence-based guidelines for depression can achieve benefits in the care of depression, e.g. in recognition, management, and clinical outcome. CONCLUSION: It was possible to implement the German Action Programme in selected care settings, and initial evaluation results suggest some improvements. The action programme provides preliminary work, materials, and results for developing a future 'Disease Management Programme' (DMP) for depression. PMID- 16260455 TI - The medical aspects of the Framingham Community Health and Tuberculosis Demonstration. PMID- 16260457 TI - Promotion to hospital consultant: regression analysis using NHS administrative data. AB - OBJECTIVES: To examine factors influencing promotion to hospital consultant. DESIGN: Multivariate logistic regression analysis of NHS administrative data between 1991 and 2000. SETTING: Hospitals in NHS Scotland. Population All registrars, senior registrars, and specialist registrars in Scotland. MAIN OUTCOME MEASURE: The proportion of doctors promoted to NHS consultant. RESULTS: Compared with doctors who graduated in Scotland, graduates from the rest of the United Kingdom and from overseas were less likely to be promoted to consultant (odds ratio 0.65, 95% confidence interval 0.52 to 0.82; and 0.37, 0.28 to 0.50, respectively). Promotion and holding an honorary contract before promotion were positively associated (1.37, 1.03 to 1.83); and the number of years since graduation (5.98 per year, 4.94 to 7.23). Women were less likely to be promoted (0.73, 0.60 to 0.90), as were doctors who worked part time (0.27, 0.17 to 0.42). Probabilities of promotion did not have a clear time trend between 1993 and 2000, and NHS boards in non-metropolitan areas of Scotland were more likely to offer promotions than NHS boards in metropolitan areas, presumably reflecting a higher gap between demand and supply in these boards. CONCLUSION: As the proportion of women in hospital medicine increases, government targets for the recruitment of consultants are unlikely to be met unless the promotion process is examined. It is unclear whether more recent reforms of the medical career structure will deal with these issues. PMID- 16260459 TI - European capacity for health promotion at the national level. PMID- 16260461 TI - No clinical evidence of hidden vCJD in UK children. AB - Between May 1997 and November 2004 this national prospective surveillance study identified 1007 children with "progressive intellectual and neurological deterioration" (PIND). In most cases specific diagnoses were made, but of 92 undiagnosed children with PIND 46 had died and only four underwent full necropsy. There was no clinical evidence of variant Creutzfeldt-Jakob disease (vCJD) in these undiagnosed cases, but without necropsy it is not possible to exclude vCJD completely. PMID- 16260462 TI - Epidemiological analysis of factors influencing an episode of exertional rhabdomyolysis in high school students. AB - BACKGROUND: An episode of rhabdomyolysis occurred after an endurance test in high school students in Taipei County in November 2003. PURPOSE: To determine the incidence, outcome, and risk factors in an episode of exertional rhabdomyolysis in high school students. STUDY DESIGN: Descriptive epidemiology study. METHODS: We enrolled all 225 high school students who had performed an endurance test. Using data from retrospective questionnaires, we estimated the incidence and assessed risk factors of exertional rhabdomyolysis among these students. Multiple logistic regression was used to determine risk factors associated with exertional rhabdomyolysis. RESULTS: The completed questionnaire was returned by 70% (157 of 225) of the students. Of these, 43.3% (68 of 157) were identified as having exertional rhabdomyolysis, and the incidence was not statistically different between male and female students (P = .49). Dark urine was noted in only 25% of the students. None of the students developed acute renal failure. The risk of exertional rhabdomyolysis was significantly higher in those students who had not exercised 1 day before the endurance test (odds ratio [OR], 6.10; 95% confidence interval [CI], 2.00-18.00) and those who had performed postexercise stretching of the legs (OR, 3.13; 95% CI, 1.28-7.69) or performed complete squats during the test (OR, 3.21; 95% CI, 1.12-10.00). There were no statistically significant differences in gender, body mass index, presence of flulike symptoms, previous exercise routine, and medication history between students with or without exertional rhabdomyolysis. CONCLUSION: Our findings suggest that exertional rhabdomyolysis is not uncommon in strenuous eccentric exercise in both men and women, but the risk of developing acute renal failure is very low. Exercise 1 day before eccentric exercise was significantly associated with a reduced risk of exertional rhabdomyolysis, whereas postexercise stretching of the involved extremities might increase the risk. PMID- 16260463 TI - Allograft reconstruction for massive, irreparable rotator cuff tears. AB - BACKGROUND: There is currently no widely accepted treatment for massive, irreparable rotator cuff tears. HYPOTHESIS: Allograft reconstruction to span the remaining defect in massive, irreparable rotator cuff tears will lead to increased functional results and will demonstrate healing of the allograft on follow-up magnetic resonance imaging studies. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: Between 1989 and 2003, 32 patients underwent allograft reconstruction of massive rotator cuff tears. University of California, Los Angeles shoulder scores were compared preoperatively and postoperatively and analyzed using paired Student t tests. In addition, 15 patients underwent postoperative magnetic resonance imaging to evaluate the structural integrity of the allograft reconstruction at the greater tuberosity insertion. RESULTS: Of 32 patients, 28 were available for review at a mean follow-up of 31.3 months; 23 of the 28 patients were satisfied with their outcome. There was 1 postoperative infection and 1 acute allograft rejection. The mean University of California, Los Angeles score increased from 12.1 preoperatively to 26.1 postoperatively (P < .001). All 15 patients evaluated with postoperative magnetic resonance imaging arthrograms demonstrated complete radiographic failure of the allograft rotator cuff reconstruction. Despite radiographic failure, the mean University of California, Los Angeles score increased from 13.2 preoperatively to 28.3 postoperatively in this subset of patients. CONCLUSION: The functional results of this reconstruction method are similar to those reported for debridement and subacromial decompression alone and are satisfactory, despite magnetic resonance imaging arthrogram evaluation demonstrating failure of the structural integrity of the allograft. Allograft reconstruction carries increased risk of infection and rejection and is technically more difficult than other less expensive treatment options with similar functional results. Therefore, we do not recommend allograft reconstruction of massive and otherwise irreparable rotator cuff tears. PMID- 16260464 TI - The relationship between quadriceps muscle force, knee flexion, and anterior cruciate ligament strain in an in vitro simulated jump landing. AB - BACKGROUND: An instrumented cadaveric knee construct was used to quantify the association between impact force, quadriceps force, knee flexion angle, and anterior cruciate ligament relative strain in simulated unipedal jump landings. HYPOTHESIS: Anterior cruciate ligament strain will correlate with impact force, quadriceps force, and knee flexion angle. STUDY DESIGN: Descriptive laboratory study. METHODS: Eleven cadaveric knees (age, 70.8 [19.3] years; 5 male; 6 female) were mounted in a custom fixture with the tibia and femur secured to a triaxial load cell. Quadriceps, hamstring, and gastrocnemius muscle forces were simulated using pretensioned steel cables (stiffness, 7 kN/cm), and the quadriceps tendon force was measured using a load cell. Mean strain on the anteromedial bundle of the anterior cruciate ligament was measured using a DVRT. With the knee in 25 degrees of flexion, the construct was vertically loaded by an impact force initially directed 4 cm posterior to the knee joint center. Tibiofemoral kinematics was measured using a 3D optoelectronic tracking system. RESULTS: The increase in anterior cruciate ligament relative strain was proportional to the increase in quadriceps force (r(2) = 0.74; P < .00001) and knee flexion angle (r(2) = 0.88; P < .00001) but was not correlated with the impact force (r(2) = 0.009; P = .08). CONCLUSION: The increase in knee flexion and quadriceps force during this simulated 1-footed landing strongly influenced the relative strain on the anteromedial bundle of the anterior cruciate ligament. CLINICAL RELEVANCE: These results suggest that even in the presence of knee flexor muscle forces, the increase in quadriceps force required to prevent the knee from flexing during landing can place the anterior cruciate ligament at risk for large strains. PMID- 16260465 TI - Replacement of the knee meniscus by a porous polymer implant: a study in dogs. AB - BACKGROUND: Meniscectomy will lead to articular cartilage degeneration in the long term. Therefore, the authors developed an implant to replace the native meniscus. HYPOTHESIS: The porous polymer meniscus implant develops into a neomeniscus and protects the cartilage from degeneration. STUDY DESIGN: Controlled laboratory study. METHODS: In a dog model, a porous polymer scaffold with optimal properties for tissue infiltration and regeneration of a neomeniscus was implanted and compared with total meniscectomy. The tissue infiltration and redifferentiation in the scaffold, the stiffness of the scaffold, and the articular cartilage degeneration were evaluated. RESULTS: Three months after implantation, the implant was completely filled with fibrovascular tissue. After 6 months, the central areas of the implant contained cartilage-like tissue with abundant collagen type II and proteoglycans in their matrix. The foreign-body reaction remained limited to a few giant cells in the implant. The compression modulus of the implant-tissue construct still differed significantly from that of the native meniscus, even at 6 months. Cartilage degeneration was observed both in the meniscectomy group and in the implant group. CONCLUSION: The improved properties of these polymer implants resulted in a faster tissue infiltration and in phenotypical differentiation into tissue resembling that of the native meniscus. However, the material characteristics of the implant need to be improved to prevent degeneration of the articular cartilage. CLINICAL RELEVANCE: The porous polymer implant developed into a polymer-tissue construct that resembled the native meniscus, and with improved gliding characteristics, this prosthesis might be a promising implant for the replacement of the meniscus. PMID- 16260467 TI - Risk factors for noncontact ankle sprains in high school football players: the role of previous ankle sprains and body mass index. AB - BACKGROUND: In a previous study, we noted a possible connection between an athlete's weight and risk of ankle sprain. HYPOTHESIS: A high body mass index and a history of a previous ankle sprain increase the risk of a subsequent noncontact sprain. STUDY DESIGN: Cohort study; Level of evidence, 2. METHODS: One hundred fifty-two athletes from 4 football teams were observed (2 varsity and 2 junior varsity). Two teams were observed for 3 seasons, and 2 teams were observed for 1 season. Before each season, body mass, height, history of previous ankle sprains, and ankle tape or brace use were recorded. RESULTS: There were 24 ankle sprains, of which 15 were noncontact inversion sprains (11 grade I, 3 grade II, 1 grade III; incidence, 1.08 per 1000 athlete-exposures). Injury incidence was higher in athletes with previous ankle injuries (2.60 vs 0.39; P < .001). Body mass index was also a risk factor (P < .05): injury incidence was 0.52 for players with a normal body mass index, 1.05 for players at risk of overweight, and 2.03 for overweight players. Injury incidence was 0.22 for normal-weight players with no previous ankle sprain compared with 4.27 for overweight players who had a previous sprain. CONCLUSION: An overweight player who had a previous ankle sprain was 19 times more likely to sustain a noncontact ankle sprain than was a normal weight player with no previous ankle sprain. CLINICAL RELEVANCE: Ankle sprain prevention strategies should be targeted at football players with a high body mass index and a history of previous ankle sprains. PMID- 16260466 TI - Kinetic comparison among the fastball, curveball, change-up, and slider in collegiate baseball pitchers. AB - BACKGROUND: Controversy exists about whether breaking pitches are more stressful than are fastballs. Previous biomechanical studies compared kinematics but not kinetics. HYPOTHESIS: Elbow and shoulder forces and torques are statistically different among the fastball, curveball, change-up, and slider. STUDY DESIGN: Descriptive laboratory study. METHODS: Twenty-one healthy collegiate pitchers were studied with a high-speed automated digitizing system. All subjects threw fastballs (n = 21), most threw curveballs (n = 20) and change-ups (n = 19), and a few threw sliders (n = 6). Wrist, elbow, and shoulder kinetics were calculated using inverse dynamics. Nine kinetic and 26 kinematic parameters were compared among the different pitch types using repeated-measures analysis of variance. RESULTS: At the shoulder, internal rotation torque, horizontal adduction torque, abduction torque, and proximal force were significantly less in the change-up than in the other 3 pitches. Shoulder horizontal adduction torque was greater in the fastball than in the curveball and slider. Shoulder proximal force was greater in the slider than in the curveball. Elbow proximal force was less in the change-up than in the other 3 pitches. Elbow varus torque was greater in the fastball and curveball than in the change-up. Elbow flexion torque was greater in the curveball than in the change-up. The curveball and change-up demonstrated kinematic differences from the fastball, consistent with previous studies. CONCLUSION: There were significant kinematic differences between the fastball and curveball but few kinetic differences. The change-up had lower joint kinetics, lower angular velocities, and different body positions than the other 3 pitch types had. Results for the slider were inconclusive because of small sample size. CLINICAL RELEVANCE: Because the resultant joint loads were similar between the fastball and curveball, this study did not indicate that either pitch was more stressful or potentially dangerous for a collegiate pitcher. The low kinetics in the change-up implies that it is the safest. PMID- 16260468 TI - Patterns of vascular and anatomical response after rotator cuff repair. AB - BACKGROUND: It has been assumed that a robust vascular response at the tendon to bone interface during rotator cuff repairs is an integral part to the healing process. There are few studies that have explored this in an in-vivo prospective fashion. PURPOSE: To prospectively characterize vascular and anatomical patterns in repaired rotator cuff tendons using Power Doppler sonography in a double blinded fashion. STUDY DESIGN: Case control study; Level of evidence, 3. METHODS: Fifty patients undergoing rotator cuff repair were enrolled: 28 mini-open, 14 open, and 8 arthroscopic repairs; 20 patients were controls. Patients underwent Power Doppler sonography at 6 weeks, 3 months, and 6 months postoperatively. Power Doppler sonography analysis examined 6 areas of the rotator cuff repair: discretely marginated intrasubstance, partial-thickness defects, full-thickness defects, focal thinning of repair, presence of bursal or joint fluid, and location of anchors. A subjective scoring system assessed blood flow in each region. RESULTS: There was a predictable, significant decrease in vascular scores after rotator cuff repair over time. The mean vascular score was 11.6 at 6 weeks, 8.3 at 3 months, 7.0 at 6 months, and 2.4 for controls. There was a significant difference (P < .05) in vascular recruitment scores between each time period, with the most robust flow at the peritendinous region. The lowest vascular score was at the anchor site or cancellous trough. Forty-eight percent of the patients had a rotator cuff repair defect postoperatively. These findings did not correlate with functional assessment and outcome at 6 months. There was no significant difference in vascular scores between the defect and no-defect groups. Mean University of California, Los Angeles; L'Insalata; and American Shoulder and Elbow Surgeons scores at 6 months were 28.6, 86.3, and 81.5, respectively. Thirty-three percent of asymptomatic controls had a rotator cuff tear that averaged 7.6 x 7.1 mm. CONCLUSION: The robust vascular response dropped with time, which is not seen in asymptomatic shoulders. Nearly half of the patients demonstrated persistent rotator cuff defects after rotator cuff repair that did not correlate with functional outcome and physical findings at 6 months. PMID- 16260469 TI - Intrinsic risk factors for the development of achilles tendon overuse injury: a prospective study. AB - BACKGROUND: Although Achilles tendon overuse injuries occur commonly, our understanding of the pathologic changes and the factors that predispose athletes to them is limited. PURPOSE: To identify measurable intrinsic risk factors for Achilles tendon overuse injuries. STUDY DESIGN: Cohort study (prognosis); Level of evidence, 2. METHODS: Sixty-nine male officer cadets followed the same 6-week basic military training. Before this training, each subject was evaluated for anthropometrical characteristics, isokinetic ankle muscle strength, ankle joint range of motion, Achilles tendon stiffness, explosive strength, and leisure and sports activity. During military training, Achilles tendon overuse injuries were registered and diagnosed by the same medical doctor. To identify the intrinsic risk factors, a multivariate analysis with the use of stepwise logistic regression was performed. The sensitivity, specificity, and cutoff values of the risk factors were evaluated by receiver operating characteristic curve analysis. RESULTS: Ten of the 69 male recruits (14.5%) sustained an Achilles tendon overuse injury diagnosed on the basis of medical history and clinical examination. Analysis revealed that male recruits with lower plantar flexor strength and increased dorsiflexion excursion were at a greater risk of Achilles tendon overuse injury. The cutoff value of the plantar flexor strength at 85% sensitivity was 50.0 N.m, with a 4.5% specificity; the cutoff value of the dorsiflexion range of motion at 85% sensitivity was 9.0 degrees , with 24.2% specificity. CONCLUSIONS: The strength of the plantar flexors and amount of dorsiflexion excursion were identified as significant predictors of an Achilles tendon overuse injury. A plantar flexor strength lower than 50.0 N.m and dorsiflexion range of motion higher than 9.0 degrees were possible thresholds for developing an Achilles tendon overuse injury. PMID- 16260470 TI - Repair of full-thickness rotator cuff tears in professional baseball players. AB - BACKGROUND: Despite the relative frequency of partial-thickness rotator cuff tears seen in baseball players, full-thickness rotator cuff tears in baseball players are uncommon. HYPOTHESIS: Return to competitive baseball is difficult after surgical treatment of a full-thickness rotator cuff tear. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: We evaluated the results of 16 professional baseball players after a mini-open repair of a full-thickness rotator cuff tear. Twelve patients were pitchers with injury to their dominant shoulders. Four patients were position players; 2 had injuries involving their dominant shoulders, and 2 had injuries to their nondominant shoulders. RESULTS: At a mean follow-up of 66.6 months for the pitchers, only 1 player (8%) was able to return to a high competitive level of baseball with no significant shoulder dysfunction after mini-open repair of a full-thickness rotator cuff tear. Of the 2 position players with mini-open repairs of the full-thickness rotator cuff tear of their dominant shoulders, 1 was able to return to professional baseball. Of the 2 position players with mini-open repairs of the full-thickness rotator cuff tear of their nondominant shoulders, both were able to return to professional baseball at the same or higher level. CONCLUSION: It is very difficult for a professional baseball pitcher to return to a competitive level of pitching after a full-thickness rotator cuff repair with a mini-open approach. PMID- 16260471 TI - A new generation of vectors with increased induction ratios by overimposing a second regulatory level by attenuation. AB - A major drawback of regulated gene expression from vectors bearing strong promoters is the associated high basal expression level. Simple regulatory systems have an intrinsic limitation in the range of induction, and attempts to mutate promoters to reduce basal expression usually result in concomitant reduction of induced levels. We have explored the possibility of reducing basal levels of gene expression while keeping induced levels intact by incorporating an additional regulatory circuit controlling a different step of the expression process. We have integrated the nasFEDCBA transcriptional attenuation system of Klebsiella oxytoca into a cascade expression circuit based on different regulatory elements of Pseudomonas putida, and also into a system based on the tac promoter, to expand their regulatory capacity. Basal expression from the promoters of these circuits was reduced by more than 10-fold by the nasF attenuator sequence while keeping the induced levels intact in the presence of the antiterminator protein, thus increasing the induction ratio by up to 1700 fold. In addition, using different combinations of regulatory elements and inducing conditions, we were able to obtain a broad range of expression levels. These vectors and the concept of their design will be very useful in regulating overproduction of heterologous proteins both at laboratory and industrial scales. PMID- 16260473 TI - Structural dynamics of cereal mitochondrial genomes as revealed by complete nucleotide sequencing of the wheat mitochondrial genome. AB - The application of a new gene-based strategy for sequencing the wheat mitochondrial genome shows its structure to be a 452 528 bp circular molecule, and provides nucleotide-level evidence of intra-molecular recombination. Single, reciprocal and double recombinant products, and the nucleotide sequences of the repeats that mediate their formation have been identified. The genome has 55 genes with exons, including 35 protein-coding, 3 rRNA and 17 tRNA genes. Nucleotide sequences of seven wheat genes have been determined here for the first time. Nine genes have an exon-intron structure. Gene amplification responsible for the production of multicopy mitochondrial genes, in general, is species specific, suggesting the recent origin of these genes. About 16, 17, 15, 3.0 and 0.2% of wheat mitochondrial DNA (mtDNA) may be of genic (including introns), open reading frame, repetitive sequence, chloroplast and retro-element origin, respectively. The gene order of the wheat mitochondrial gene map shows little synteny to the rice and maize maps, indicative that thorough gene shuffling occurred during speciation. Almost all unique mtDNA sequences of wheat, as compared with rice and maize mtDNAs, are redundant DNA. Features of the gene based strategy are discussed, and a mechanistic model of mitochondrial gene amplification is proposed. PMID- 16260472 TI - Information theory based T7-like promoter models: classification of bacteriophages and differential evolution of promoters and their polymerases. AB - Molecular information theory was used to create sequence logos and promoter models for eight phages of the T7 group: T7, phiA1122, T3, phiYeO3-12, SP6, K1-5, gh-1 and K11. When these models were used to scan the corresponding genomes, a significant gap in the individual information distribution was observed between functional promoter sites and other sequences, suggesting that the models can be used to identify new T7-like promoters. When a combined 76-site model was used to scan the eight phages, 108 of the total 109 promoters were found, while none were found for other T7-like phages, phiKMV, P60, VpV262, SIO1, PaP3, Xp10, P-SSP7 and Ppu40, indicating that these phages do not belong to the T7 group. We propose that the T7-like transcription system, which consists of a phage-specific RNA polymerase and a set of conserved T7-like promoters, is a hallmark feature of the T7 group and can be used to classify T7-like phages. Phylogenetic trees of the T7 like promoter models and their corresponding RNA polymerases are similar, suggesting that the eight phages of the T7 group can be classified into five subgroups. However the SP6-like polymerases have apparently diverged from other polymerases more than their promoters have diverged from other promoters. PMID- 16260474 TI - Identification of RecQL1 as a Holliday junction processing enzyme in human cell lines. AB - Homologous recombination provides an effective way to repair DNA double-strand breaks (DSBs) and is required for genetic recombination. During the process of homologous recombination, a heteroduplex DNA structure, or a 'Holliday junction' (HJ), is formed. The movement, or branch migration, of this junction is necessary for recombination to proceed correctly. In prokaryotes, the RecQ protein or the RuvA/RuvB protein complex can promote ATP-dependent branch migration of Holliday junctions. Much less is known about the processing of Holliday junctions in eukaryotes. Here, we identify RecQL1 as a predominant ATP-dependent, HJ branch migrator present in human nuclear extracts. A reduction in the level of RecQL1 induced by RNA interference in HeLa cells leads to an increase in sister chromatid exchange. We propose that RecQL1 is involved in the processing of Holliday junctions in human cells. PMID- 16260475 TI - Detection of low-level promoter activity within open reading frame sequences of Escherichia coli. AB - The search for promoters has largely been confined to sequences upstream of open reading frames (ORFs) or stable RNA genes. Here we used a cloning approach to discover other potential promoters in Escherichia coli. Chromosomal fragments of approximately 160 bp were fused to a promoterless lacZ reporter gene on a multi copy plasmid. Eight clones were deliberately selected for high activity and 105 clones were selected at random. All eight of the high-activity clones carried promoters that were located upstream of an ORF. Among the randomly-selected clones, 56 had significantly elevated activity. Of these, 7 had inserts which also mapped upstream of an ORF, while 49 mapped within or downstream of ORFs. Surprisingly, the eight promoters selected for high activity matched the canonical sigma70 -35 and -10 sequences no better than sequences from the randomly-selected clones. For six of the nine most active sequences with orientations opposite to that of the ORF, chromosomal expression was detected by RT-PCR, but defined transcripts were not detected by northern analysis. Our results indicate that the E.coli chromosome carries numerous -35 and -10 sequences with weak promoter activity but that most are not productively expressed because other features needed to enhance promoter activity and transcript stability are absent. PMID- 16260477 TI - Editorial: a more compassionate approach to punishment? PMID- 16260476 TI - The POU protein Oct-6 is a nucleocytoplasmic shuttling protein. AB - Like many POU domain proteins, Oct-6 plays important roles during vertebrate development. In accord with its function as a transcriptional regulator during neurogenesis and myelination, Oct-6 is predominantly found in the nucleus. Nuclear import is mediated by a nuclear localization signal at the N-terminal end of the POU homeodomain. Here we show, that Oct-6 in addition contains a nuclear export signal so that Oct-6 is able to shuttle constantly between nucleus and cytoplasm. This nuclear export signal is also localized in the POU homeodomain as part of helix 2 and the connecting loop to DNA recognition helix 3. It conforms to the consensus of hydrophobic leucine-rich export sequences and mediates export from the nucleus via CRM1/Exp1. Several amino acid substitutions or insertions that inactivate this nuclear export sequence, reduce DNA-binding of Oct-6 to its octamer recognition element slighty, but interfere strongly with Oct-6-dependent transcriptional activation, thus arguing that nuclear export and nucleocytoplasmic shuttling are essential aspects of Oct-6 function. Importantly, the nuclear export signal identified for Oct-6 is conserved in most, if not all other vertebrate POU proteins. Nuclear export might therefore be of general relevance for POU protein function throughout development. PMID- 16260478 TI - Life on the atoll: Singapore ecology as a neglected dimension of social order. AB - Based on field interviews in Singapore in 1985, 1997, and 2003, this article addresses the issue of how island ecology helps explain the remarkable low rates of crime that are often attributed mainly to cultural and government policies. Understanding crime and control on this most densely populated Southeast Asian atoll must begin with how people are dispersed over the limited spatial area. Ecology also influences how styles of some crimes are defined and controlled. Several of Donald Black's propositions are given further consideration. PMID- 16260479 TI - Isolation and normlessness: attitudinal comparisons of adolescent sex offenders, juvenile offenders, and nondelinquents. AB - The authors explored attitudinal differences among adolescent male sex offenders, juvenile delinquents, and nondelinquent youth based on three variables drawn from integrated delinquency theory: conventional attitudes, normlessness, and social isolation. Consistent with previous juvenile delinquency studies, the results indicate no differences among the three groups on conventional attitudes. With respect to normlessness, both the sex offenders and juvenile delinquent groups demonstrated more school normlessness than did nondelinquent youths, and adolescent sex offenders showed greater peer normlessness than did either nondelinquent youths or juvenile delinquents. Examination of perceived social isolation among the three groups indicates that sex offenders consistently perceived themselves as more isolated than other youths with their families, in their school, and among their peers. These results suggest that interpersonal factors, in addition to a lack of social controls and normlessness, are associated with sexually inappropriate behavior. PMID- 16260480 TI - A comparison of the male customers of female street prostitutes with national samples of men. AB - Previous research on customers of prostitutes has relied on small samples and qualitative interviews. Conceptions of customers have tended toward either the "every man" perspective, which implies customers are no different than other men, or the "peculiar man" perspective, which implies customers are characterized by stark differences or psychological inadequacies. This study compares a large sample of men (N = 1672) arrested for trying to hire street prostitutes with nationally representative samples of men. Customers were less likely to be married, less likely to be happily married if married, and more likely to report being unhappy in general than men in the national samples. Customers also expressed greater sexual liberalism and reported thinking about sex, masturbating, and participating in other aspects of the sex industry more frequently than men in general. Most differences were small, indicating customers as a category differ from other men in degree rather than quality. PMID- 16260481 TI - A Rorschach investigation of incarcerated female offenders with antisocial personality disorder. AB - Although male psychopathy has been linked to histrionic, narcissistic, and antisocial personality disorders (ASPD), less is known about female psychopathy. The Psychopathy Checklist-Revised (PCL-R) and the Rorschach were used to explore the personality functioning of 45 incarcerated female offenders with ASPD delineated by their psychopathy level. Psychopaths (PCL-R > or = 30) and nonpsychopaths (PCL-R < 24) were compared on Rorschach measures of self perception, interpersonal relatedness, and reality testing. Compared to female offenders with ASPD who were nonpsychopathic, female offenders with ASPD who were psychopathic exhibited marked disturbances in self-perception, interpersonal relatedness, and reality testing. Our findings highlight the heterogeneity of the ASPD diagnosis in women, support the utility of the psychopathy construct with female offenders, and implicate important differences between men and women with ASPD. These gender differences have relevance to the evaluation (PCL-R scoring) and treatment of female offenders. Our findings are discussed within the context of the female psychopath's hypothesized hysterical character style. PMID- 16260482 TI - Cross-validation of the self-appraisal questionnaire: a tool for assessing violent and nonviolent recidivism with female offenders. AB - The Self-Appraisal Questionnaire (SAQ) is a 72-item self-report measure designed to predict violent and nonviolent recidivism among adult male criminal offenders. It was administered to 91 female offenders incarcerated in Pennsylvania and 183 incarcerated in Singapore correctional systems. Results indicated that the SAQ has sound psychometric properties, with acceptable reliability and concurrent and predictive validity for assessing violent and nonviolent recidivism. There were no significant differences between the scores of African American and Asian offenders and the responses of the White offenders. Similar to the findings from male offenders, the present results provide some support for the validity of the SAQ in the prediction of violent and nonviolent recidivism risk among White, African American, and Asian female offenders. PMID- 16260483 TI - Expressed needs and behavioral risk factors of HIV-positive inmates. AB - Considerable research has found inmates to be at high risk for HIV/AIDS. Furthermore, research has shown additional HIV/AIDS threats faced by females in the general population and especially by those incarcerated. Behavioral interventions and educational programs have been developed based on these studies. However, few empirical studies have examined the self-expressed needs of HIV-positive inmates or the degree to which inmates' needs are gender specific. This pilot study compares the needs of HIV-positive male and female jail detainees. Results illustrate surprisingly few differences between men and women and their HIV-related needs. The primary need identified for both males and females was postrelease housing. Somewhat unexpectedly, HIV treatment and care ranked low on the list of needs. The implications of these finds are discussed. PMID- 16260484 TI - Incarcerated clients' perceptions of therapeutic change in substance abuse treatment: a 4-year case study. AB - Therapeutic communities (TCs) are a primary approach for treating substance abusers in the criminal justice system. The transformation from residential treatment to a TC marked a significant change in the service delivery and organizational culture for one in-prison substance abuse treatment program. This longitudinal case study examines how clients perceived their treatment as a result of treatment and organizational changes. This is important because the scientific literature shows that client perceptions of their treatment impacts treatment retention and the benefit they gain from treatment. Data that were collected over 5 years are presented. Implications, including how the program's interim therapeutic milieu eased the transition to a therapeutic community, are discussed. PMID- 16260485 TI - The effects of cognitive-behavioral treatment for forensic inpatients. AB - The effects of an intramural cognitive-behavioral treatment for forensic inpatients with personality disorders in a high-security hospital were examined. Treatment was aimed at modifying maladaptive coping and social skills, at enhancing social awareness, at reducing egoistic and oppositional behaviors, and at reducing psychological complaints. The patients, who all had committed serious crimes (violence, arson, sexual offences), participated voluntarily in the study. A total of 39 patients started the study, but during the course of the study, several patients dropped out because of several reasons. Patients as a group showed significant improvements over time on psychopathological symptoms, personality traits, and coping. A significant decrease of oppositional behaviors was reported by the staff. Though the patients improved well at the group level, only a minority of patients showed reliable change over time at the individual level. The meaning of the results in relation to treatment are discussed. PMID- 16260486 TI - RNA-associated autoantigens activate B cells by combined B cell antigen receptor/Toll-like receptor 7 engagement. AB - Previous studies (Leadbetter, E.A., I.R. Rifkin, A.H. Hohlbaum, B. Beaudette, M.J. Shlomchik, and A. Marshak-Rothstein. 2002. Nature. 416:603-607; Viglianti, G.A., C.M. Lau, T.M. Hanley, B.A. Miko, M.J. Shlomchik, and A. Marshak-Rothstein. 2003. Immunity. 19:837-847) established the unique capacity of DNA and DNA associated autoantigens to activate autoreactive B cells via sequential engagement of the B cell antigen receptor (BCR) and Toll-like receptor (TLR) 9. We demonstrate that this two-receptor paradigm can be extended to the BCR/TLR7 activation of autoreactive B cells by RNA and RNA-associated autoantigens. These data implicate TLR recognition of endogenous ligands in the response to both DNA- and RNA-associated autoantigens. Importantly, the response to RNA-associated autoantigens was markedly enhanced by IFN-alpha, a cytokine strongly linked to disease progression in patients with systemic lupus erythematosus (SLE). As further evidence that TLRs play a key role in autoantibody responses in SLE, we found that autoimmune-prone mice, lacking the TLR adaptor protein MyD88, had markedly reduced chromatin, Sm, and rheumatoid factor autoantibody titers. PMID- 16260487 TI - Characterization of marginal zone B cell precursors. AB - Selection of recently formed B cells into the follicular or marginal zone (MZ) compartments is proposed to occur by way of proliferative intermediates expressing high levels of CD21/35 and CD23. However, we show that CD21/35(high) CD23(+) splenocytes are not enriched for proliferative cells, and do not contribute substantially to the generation of follicular B cells. Instead, ontogenic relationships, steady-state labeling kinetics, and adoptive transfer experiments suggest that CD21/35(high) CD23(+) splenocytes serve primarily as precursors for MZ B cells, although their developmental potential seems to be broader and is influenced by environmental cues that are associated with lymphopenia. Furthermore, CD21/35(high) CD23(+) splenocytes share several key functional characteristics with MZ B cells, including their capacity to trap T independent antigen and a heightened proliferative response to LPS. These observations challenge previous models of peripheral B cell maturation, and suggest that MZ B cells develop by way of CD21/35(high) CD23(+) intermediates. PMID- 16260488 TI - The insulin A-chain epitope recognized by human T cells is posttranslationally modified. AB - The autoimmune process that destroys the insulin-producing pancreatic beta cells in type 1 diabetes (T1D) is targeted at insulin and its precursor, proinsulin. T cells that recognize the proximal A-chain of human insulin were identified recently in the pancreatic lymph nodes of subjects who had T1D. To investigate the specificity of proinsulin-specific T cells in T1D, we isolated human CD4(+) T cell clones to proinsulin from the blood of a donor who had T1D. The clones recognized a naturally processed, HLA DR4-restricted epitope within the first 13 amino acids of the A-chain (A1-13) of human insulin. T cell recognition was dependent on the formation of a vicinal disulfide bond between adjacent cysteine residues at A6 and A7, which did not alter binding of the peptide to HLA DR4. CD4(+) T cell clones that recognized this epitope were isolated from an HLA DR4(+) child with autoantibodies to insulin, and therefore, at risk for T1D, but not from two healthy HLA DR4(+) donors. We define for the first time a novel posttranslational modification that is required for T cell recognition of the insulin A-chain in T1D. PMID- 16260489 TI - Live attenuated yellow fever 17D infects human DCs and allows for presentation of endogenous and recombinant T cell epitopes. AB - The yellow fever (YF) 17D vaccine is one of the most successful live attenuated vaccines available. A single immunization induces both long-lasting neutralizing antibody and YF-specific T cell responses. Surprisingly, the mechanism for this robust immunity has not been addressed. In light of several recent reports suggesting flavivirus interaction with dendritic cells (DCs), we investigated the mechanism of YF17D interaction with DCs and the importance of this interaction in generating T cell immunity. Our results show that YF17D can infect immature and mature human DCs. Viral entry is Ca(2+) dependent, but it is independent of DC SIGN as well as multiple integrins expressed on the DC surface. Similar to infection of cell lines, YF infection of immature DCs is cytopathic. Although infection itself does not induce DC maturation in vitro, TNF-alpha-induced maturation protects DCs from YF-induced cytopathogenicity. Furthermore, we show that DCs infected with YF17D or YF17D carrying a recombinant epitope can process and present antigens for CD8(+) T cell stimulation. These findings offer insight into the immunologic mechanisms associated with the highly capable YF17D vaccine that may guide effective vaccine design. PMID- 16260490 TI - A role for dual viral hits in causation of subacute sclerosing panencephalitis. AB - Subacute sclerosing panencephalitis (SSPE) is a progressive fatal neurodegenerative disease associated with persistent infection of the central nervous system (CNS) by measles virus (MV), biased hypermutations of the viral genome affecting primarily the matrix (M) gene with the conversion of U to C and A to G bases, high titers of antibodies to MV, and infiltration of B cells and T cells into the CNS. Neither the precipitating event nor biology underlying the MV infection is understood, nor is their any satisfactory treatment. We report the creation of a transgenic mouse model that mimics the cardinal features of SSPE. This was achieved by initially infecting mice expressing the MV receptor with lymphocytic choriomeningitis virus Cl 13, a virus that transiently suppressed their immune system. Infection by MV 10 days later resulted in persistent MV infection of neurons. Analysis of brains from infected mice showed the biased U to C hypermutations in the MV M gene and T and B lymphocyte infiltration. These sera contained high titers of antibodies to MV. Thus, a small animal model is now available to both molecularly probe the pathogenesis of SSPE and to test a variety of therapies to treat the disease. PMID- 16260491 TI - Protection from Alzheimer's-like disease in the mouse by genetic ablation of inducible nitric oxide synthase. AB - Brains from subjects who have Alzheimer's disease (AD) express inducible nitric oxide synthase (iNOS). We tested the hypothesis that iNOS contributes to AD pathogenesis. Immunoreactive iNOS was detected in brains of mice with AD-like disease resulting from transgenic expression of mutant human beta-amyloid precursor protein (hAPP) and presenilin-1 (hPS1). We bred hAPP-, hPS1-double transgenic mice to be iNOS(+/+) or iNOS(-/-), and compared them with a congenic WT strain. Deficiency of iNOS substantially protected the AD-like mice from premature mortality, cerebral plaque formation, increased beta-amyloid levels, protein tyrosine nitration, astrocytosis, and microgliosis. Thus, iNOS seems to be a major instigator of beta-amyloid deposition and disease progression. Inhibition of iNOS may be a therapeutic option in AD. PMID- 16260492 TI - Loss of splicing factor ASF/SF2 induces G2 cell cycle arrest and apoptosis, but inhibits internucleosomal DNA fragmentation. AB - ASF/SF2 is an SR protein splicing factor that participates in constitutive and alternative pre-mRNA splicing and is essential for cell viability. Using a genetically modified chicken B-cell line, DT40-ASF, we now show that ASF/SF2 inactivation results in a G2-phase cell cycle arrest and subsequent programmed cell death. However, although several hallmarks of apoptosis are apparent, internucleosomal DNA fragmentation was not detected. Furthermore, inactivation of ASF/SF2 also blocks DNA fragmentation normally induced by a variety of apoptotic stimuli. Notably, mRNA encoding the inhibitor of caspase-activated DNase-L (ICAD L), which acts as an inhibitor as well as a chaperone of caspase-activated DNase (CAD), decreased in abundance, whereas the level of mRNA encoding ICAD-S, which has only inhibitory activity, increased upon ASF/SF2 depletion. Strikingly, expression of appropriate levels of exogenous human ICAD-L restored apoptotic DNA laddering in ASF/SF2-depleted cells. These results not only indicate that loss of an SR protein splicing factor can induce cell cycle arrest and apoptosis, but also illustrate the important role of ICAD and its regulation by alternative splicing in the process of apoptotic DNA fragmentation. PMID- 16260495 TI - Morphogen gradient interpretation by a regulated trafficking step during ligand receptor transduction. AB - Morphogen gradients are important in early development, but how cells recognize their position in such a gradient is not well understood. Cells need to correctly interpret a morphogen concentration when the morphogen is no longer present in the extracellular medium. This memory of morphogen exposure is necessary for correct cell fate decisions in the changing morphogen gradient concentration in an embryo. Our results demonstrate that a previously unrecognized step in gradient interpretation is a temporal stop that arrests the progression of a ligand-receptor complex between internalization and lysosomal destruction. Signaling continues during this arrested progression, which constitutes the basis of memory of morphogen concentration. We show that prolonged signaling requires Dynamin-dependent internalization of the complex. Rab5QL- and Rab7QL-mediated increases in the speed of the endo-lysosomal progression do not affect memory. In contrast, memory is abolished by increasing the targeting of receptors to the lysosome through expression of the Smad7/Smurf2 ubiquitin ligase. We conclude that the basis for memory is the long-lasting residence of a signaling complex in the endo-lysosomal pathway. The regulated duration of this step helps to determine the choice of gene expression resulting from gradient interpretation. PMID- 16260493 TI - TAK1, but not TAB1 or TAB2, plays an essential role in multiple signaling pathways in vivo. AB - TGF-beta-activated kinase 1 (TAK1), a member of the MAPKKK family, is thought to be a key modulator of the inducible transcription factors NF-kappaB and AP-1 and, therefore, plays a crucial role in regulating the genes that mediate inflammation. Although in vitro biochemical studies have revealed the existence of a TAK1 complex, which includes TAK1 and the adapter proteins TAB1 and TAB2, it remains unclear which members of this complex are essential for signaling. To analyze the function of TAK1 in vivo, we have deleted the Tak1 gene in mice, with the resulting phenotype being early embryonic lethality. Using embryonic fibroblasts lacking TAK1, TAB1, or TAB2, we have found that TNFR1, IL-1R, TLR3, and TLR4-mediated NF-kappaB and AP-1 activation are severely impaired in Tak1(m/m) cells, but they are normal in Tab1(-/-) and Tab2(-/-) cells. In addition, Tak1(m/m) cells are highly sensitive to TNF-induced apoptosis. TAK1 mediates IKK activation in TNF-alpha and IL-1 signaling pathways, where it functions downstream of RIP1-TRAF2 and MyD88-IRAK1-TRAF6, respectively. However, TAK1 is not required for NF-kappaB activation through the alternative pathway following LT-beta signaling. In the TGF-beta signaling pathway, TAK1 deletion leads to impaired NF-kappaB and c-Jun N-terminal kinase (JNK) activation without impacting Smad2 activation or TGF-beta-induced gene expression. Therefore, our studies suggests that TAK1 acts as an upstream activating kinase for IKKbeta and JNK, but not IKKalpha, revealing an unexpectedly specific role of TAK1 in inflammatory signaling pathways. PMID- 16260494 TI - The tumor suppressors Ink4c and p53 collaborate independently with Patched to suppress medulloblastoma formation. AB - Recurrent genetic alterations in human medulloblastoma (MB) include mutations in the sonic hedgehog (SHH) signaling pathway and TP53 inactivation (approximately 25% and 10% of cases, respectively). However, mouse models of MB, regardless of their initiating lesions, generally depend upon p53 inactivation for rapid onset and high penetrance. The gene encoding the cyclin-dependent kinase inhibitor p18(Ink4c) is transiently expressed in mouse cerebellar granule neuronal precursor cells (GNPs) as they exit the cell division cycle and differentiate. Coinactivation of Ink4c and p53 provided cultured GNPs with an additive proliferative advantage, either in the presence or absence of Shh, and induced MB with low penetrance but with greatly increased incidence following postnatal irradiation. In contrast, mice lacking one or two functional Ink4c alleles and one copy of Patched (Ptc1) encoding the Shh receptor rapidly developed MBs that retained wild-type p53. In tumor cells purified from double heterozygotes, the wild-type Ptc1 allele, but not Ink4c, was inactivated. Therefore, when combined with Ptc1 mutation, Ink4c is haploinsufficient for tumor suppression. Methylation of INK4C (CDKN2C) was observed in four of 23 human MBs, and p18(INK4C) protein expression was extinguished in 14 of 73 cases. Hence, p18(INK4C) loss may contribute to MB formation in children. PMID- 16260496 TI - Phosphorylation by Cdk1 induces Plk1-mediated vimentin phosphorylation during mitosis. AB - Several kinases phosphorylate vimentin, the most common intermediate filament protein, in mitosis. Aurora-B and Rho-kinase regulate vimentin filament separation through the cleavage furrow-specific vimentin phosphorylation. Cdk1 also phosphorylates vimentin from prometaphase to metaphase, but its significance has remained unknown. Here we demonstrated a direct interaction between Plk1 and vimentin-Ser55 phosphorylated by Cdk1, an event that led to Plk1 activation and further vimentin phosphorylation. Plk1 phosphorylated vimentin at approximately 1 mol phosphate/mol substrate, which partly inhibited its filament forming ability, in vitro. Plk1 induced the phosphorylation of vimentin-Ser82, which was elevated from metaphase and maintained until the end of mitosis. This elevation followed the Cdk1-induced vimentin-Ser55 phosphorylation, and was impaired by Plk1 depletion. Mutational analyses revealed that Plk1-induced vimentin-Ser82 phosphorylation plays an important role in vimentin filaments segregation, coordinately with Rho-kinase and Aurora-B. Taken together, these results indicated a novel mechanism that Cdk1 regulated mitotic vimentin phosphorylation via not only a direct enzyme reaction but also Plk1 recruitment to vimentin. PMID- 16260497 TI - Traffic of Kv4 K+ channels mediated by KChIP1 is via a novel post-ER vesicular pathway. AB - The traffic of Kv4 K+ channels is regulated by the potassium channel interacting proteins (KChIPs). Kv4.2 expressed alone was not retained within the ER, but reached the Golgi complex. Coexpression of KChIP1 resulted in traffic of the channel to the plasma membrane, and traffic was abolished when mutations were introduced into the EF-hands with channel captured on vesicular structures that colocalized with KChIP1(2-4)-EYFP. The EF-hand mutant had no effect on general exocytic traffic. Traffic of Kv4.2 was coat protein complex I (COPI)-dependent, but KChIP1-containing vesicles were not COPII-coated, and expression of a GTP loaded Sar1 mutant to block COPII function more effectively inhibited traffic of vesicular stomatitis virus glycoprotein (VSVG) than did KChIP1/Kv4.2 through the secretory pathway. Therefore, KChIP1seems to be targeted to post-ER transport vesicles, different from COPII-coated vesicles and those involved in traffic of VSVG. When expressed in hippocampal neurons, KChIP1 co-distributed with dendritic Golgi outposts; therefore, the KChIP1 pathway could play an important role in local vesicular traffic in neurons. PMID- 16260498 TI - Excess of Gbetae over Gqalphae in vivo prevents dark, spontaneous activity of Drosophila photoreceptors. AB - Drosophila melanogaster photoreceptor cells are capable of detecting single photons. This utmost sensitivity is critically dependent on the maintenance of an exceedingly low, dark, spontaneous activity of photoreceptor cells. However, the underlying mechanisms of this hallmark of phototransduction are not fully understood. An analysis of the Drosophila visual heterotrimeric (alphabetagamma) Gq protein revealed that wild-type Drosophila flies have about a twofold excess of Gbeta over Galpha subunits of the visual Gq protein. Studies of Gbetae mutants in which the excess of Gbeta was genetically eliminated showed dramatic dark, spontaneous activity of the photoreceptor cells, whereas concurrent genetic reduction of the Galpha subunit, which restored the excess of Gbeta, abolished this effect. These results indicate that an excess of Gbeta over Galpha is a strategy used in vivo for the suppression of spontaneous activity, thereby yielding a high signal to noise ratio, which is characteristic of the photoreceptor light response. This mechanism could be relevant to the regulation of G protein signaling in general. PMID- 16260499 TI - Localization of MMR proteins on meiotic chromosomes in mice indicates distinct functions during prophase I. AB - Mammalian MutL homologues function in DNA mismatch repair (MMR) after replication errors and in meiotic recombination. Both functions are initiated by a heterodimer of MutS homologues specific to either MMR (MSH2-MSH3 or MSH2-MSH6) or crossing over (MSH4-MSH5). Mutations of three of the four MutL homologues (Mlh1, Mlh3, and Pms2) result in meiotic defects. We show herein that two distinct complexes involving MLH3 are formed during murine meiosis. The first is a stable association between MLH3 and MLH1 and is involved in promoting crossing over in conjunction with MSH4-MSH5. The second complex involves MLH3 together with MSH2 MSH3 and localizes to repetitive sequences at centromeres and the Y chromosome. This complex is up-regulated in Pms2-/- males, but not females, providing an explanation for the sexual dimorphism seen in Pms2-/- mice. The association of MLH3 with repetitive DNA sequences is coincident with MSH2-MSH3 and is decreased in Msh2-/- and Msh3-/- mice, suggesting a novel role for the MMR family in the maintenance of repeat unit integrity during mammalian meiosis. PMID- 16260500 TI - Drosophila melanogaster Cad99C, the orthologue of human Usher cadherin PCDH15, regulates the length of microvilli. AB - Actin-based protrusions can form prominent structures on the apical surface of epithelial cells, such as microvilli. Several cytoplasmic factors have been identified that control the dynamics of actin filaments in microvilli. However, it remains unclear whether the plasma membrane participates actively in microvillus formation. In this paper, we analyze the function of Drosophila melanogaster cadherin Cad99C in the microvilli of ovarian follicle cells. Cad99C contributes to eggshell formation and female fertility and is expressed in follicle cells, which produce the eggshells. Cad99C specifically localizes to apical microvilli. Loss of Cad99C function results in shortened and disorganized microvilli, whereas overexpression of Cad99C leads to a dramatic increase of microvillus length. Cad99C that lacks most of the cytoplasmic domain, including potential PDZ domain-binding sites, still promotes excessive microvillus outgrowth, suggesting that the amount of the extracellular domain determines microvillus length. This study reveals Cad99C as a critical regulator of microvillus length, the first example of a transmembrane protein that is involved in this process. PMID- 16260502 TI - Cytoplasmic dynein nomenclature. AB - A variety of names has been used in the literature for the subunits of cytoplasmic dynein complexes. Thus, there is a strong need for a more definitive consensus statement on nomenclature. This is especially important for mammalian cytoplasmic dyneins, many subunits of which are encoded by multiple genes. We propose names for the mammalian cytoplasmic dynein subunit genes and proteins that reflect the phylogenetic relationships of the genes and the published studies clarifying the functions of the polypeptides. This nomenclature recognizes the two distinct cytoplasmic dynein complexes and has the flexibility to accommodate the discovery of new subunits and isoforms. PMID- 16260503 TI - Expert witnesses: perpetuating a flawed system. PMID- 16260501 TI - How does the TOM complex mediate insertion of precursor proteins into the mitochondrial outer membrane? AB - A multisubunit translocase of the outer mitochondrial membrane (TOM complex) mediates both the import of mitochondrial precursor proteins into the internal compartments of the organelle and the insertion of proteins residing in the mitochondrial outer membrane. The proposed beta-barrel structure of Tom40, the pore-forming component of the translocase, raises the question of how the apparent uninterrupted beta-barrel topology can be compatible with a role of Tom40 in releasing membrane proteins into the lipid core of the bilayer. In this review, I discuss insertion mechanisms of proteins into the outer membrane and present alternative models based on the opening of a multisubunit beta-barrel TOM structure or on the interaction of outer membrane precursors with the outer face of the Tom40 beta-barrel structure. PMID- 16260504 TI - Embracing change in obstetrics and gynecology. PMID- 16260505 TI - Forceps compared with vacuum: rates of neonatal and maternal morbidity. AB - OBJECTIVE: To compare perinatal outcomes between forceps- and vacuum-assisted deliveries. Our hypothesis was that the force vectors achieved in forceps delivery will lead to fewer shoulder dystocias, but greater perineal lacerations. METHODS: This was a retrospective cohort study of 4,120 term, cephalic, singleton, nonrotational operative vaginal deliveries at a single institution. Outcomes examined included rates of neonatal trauma, shoulder dystocia, and perineal lacerations. Potential confounders, including maternal age, birthweight, ethnicity, parity, station at delivery, episiotomy, attending physician, anesthesia, and length of labor, were controlled for using multivariate logistic regression. RESULTS: Among the 2,075 (50.4%) forceps- and 2,045 (49.6%) vacuum assisted deliveries, the rate of shoulder dystocia was lower among women undergoing forceps delivery (1.5% compared with 3.5%, P < .001), as was the rate of cephalohematoma (4.5% compared with 14.8%, P < .001), whereas the rate of third- or fourth-degree perineal laceration was higher (36.9% compared with 26.8%, P < .001). These differences in perinatal complications persisted when controlling for the confounders listed above. The adjusted odds ratio for shoulder dystocia was 0.34 (95% confidence interval [CI] 0.20-0.57), for cephalohematoma was 0.25 (95% CI 0.19-0.33), and for third- or fourth-degree lacerations was 1.79 (95% CI 1.52-2.10) when comparing forceps to vacuum. CONCLUSION: Vacuum-assisted vaginal birth is more often associated with shoulder dystocia and cephalohematoma. Forceps delivery is more often associated with third- and fourth-degree perineal lacerations. These differences in complications rates should be considered among other factors when determining the optimal mode of delivery. LEVEL OF EVIDENCE: II-2. PMID- 16260506 TI - Risk factors for obstetric brachial plexus palsy among neonates delivered by vacuum extraction. AB - OBJECTIVE: The risk of obstetric brachial plexus palsy (OBPP) is increased in infants delivered instrumentally. The aim of this study was to identify risk factors for OBPP and to evaluate the association between possible risk factors linked to the duration of the vacuum extraction procedure and the subsequent risk. METHODS: A population-based retrospective design was adopted. Using a national registry of operative vaginal deliveries linked to the Medical Birth Registry in Sweden, we evaluated by univariate and multiple logistic regression analyses the risk factors for OBPP in 13,716 women delivered by vacuum extraction. The variables assessed in the multiple logistic regression analysis were shoulder dystocia, fetal birth weight of 3,999 g or greater, fundal pressure, number of tractions, vacuum application time, parity, vacuum silicone cup, epidural anesthesia, and fetal head at the level of the ischial spines at vacuum application time. RESULTS: Obstetric brachial plexus palsy was recorded in 153 (1.1%) infants. The following variables increased significantly the risk of OBPP in the newborn: shoulder dystocia (odds ratio 16.0; 95% confidence interval 8.9-28.7), fetal birth weight of 3,999 g or greater (7.1; 4.8-10.5), and administration of fundal pressure (1.6; 1.1-2.3). The probability of the risk of OBPP in vacuum-assisted deliveries increased in relation to vacuum extraction time (minutes). CONCLUSION: Shoulder dystocia in the setting of vacuum extraction is a prominent risk factor for OBPP in the newborn. The risk of OBPP increases with the time required for vacuum extraction. LEVEL OF EVIDENCE: II-3. PMID- 16260507 TI - Magnetic resonance imaging pelvimetry and the prediction of labor dystocia. AB - OBJECTIVE: To study whether magnetic resonance imaging (MRI) pelvimetry has the ability to identify those women who require cesarean delivery for labor dystocia. METHODS: From July 2003 to April 2004, nulliparous women scheduled for a labor induction for prolonged pregnancy (42 weeks) were asked to participate in a pelvimetry study. Those who consented underwent fast-acquisition MRI that included two 90-second acquisitions to evaluate fetal biometry and volumetry and maternal pelvimetry, including novel measurements of pelvic bony and soft tissue volumes as determined by MRI. Information about each patient's pregnancy, labor course, and neonatal outcome was prospectively collected. Pelvimetry results for those women undergoing operative delivery for labor dystocia were compared with those who did not. Single fetal and maternal pelvic measurements, as well as ratios of both, were analyzed. In addition, previously described radiographic pelvimetry techniques and formulas to predict dystocia were used. RESULTS: One hundred one women underwent MRI, and 22 of these underwent cesarean delivery for dystocia. No single fetal measurement was statistically associated with dystocia. Several maternal pelvic measures, fetal-to-maternal ratios, and previously reported pelvimetric techniques were significantly associated with dystocia. The ratio of magnetic resonance (MR) fetal head volume to pelvic soft tissue volume had statistical significance (P = .04). Receiver operator characteristic curves were developed for the different measurements, ratios, and formulas studied to assess whether any of the techniques could accurately predict labor dystocia requiring operative delivery. The area under the curve values ranged from 0.6 to 0.8, with the ratio of MR head volume to pelvic soft tissue being 0.7. These values suggest that MRI can identify those women at greatest risk for dystocia, but it cannot with accuracy predict which ones will require a cesarean. CONCLUSION: We found significant associations with MRI pelvimetry and labor dystocia, but MRI was not a significant improvement over previously described pelvimetric techniques. LEVEL OF EVIDENCE: II-3. PMID- 16260508 TI - The effect of fetal number on the development of hypertensive conditions of pregnancy. AB - OBJECTIVE: To estimate the incidence and effect of pregnancy-related hypertensive conditions on multiple gestations. METHODS: Women with 1-4 fetuses enrolled in an outpatient perinatal services program at 28 or more weeks of gestation were identified. Those without a prior diagnosis of hypertension at enrollment and who delivered at more than 28 weeks of gestation were included in the analysis. The incidence of all pregnancy-related hypertensive conditions, diagnosis of severe hypertensive conditions (hemolysis, elevated liver enzymes and low platelets syndrome; disseminated intravascular coagulation; eclampsia; low platelets; renal failure; and abruption), and interventional delivery related to hypertension were estimated, and compared according to fetal number. RESULTS: Data were analyzed for 34,374 singleton, twin, triplet, and quadruplet gestations. The incidence of pregnancy-related hypertensive conditions increased with multifetal gestations as compared with singletons (12.7-19.6% for multifetal gestations compared with 6.5% for singletons, P < .001). The incidence of severe pregnancy-related hypertensive conditions was significantly increased in twin (1.6%) and triplet (3.1%) gestations as compared with singletons (0.5%, P < .001). Quadruplet pregnancies were not significantly higher than triplet gestations for these complications. The need for early delivery related to hypertension was greater with increasing fetal number through triplet gestations. By logistic regression, higher fetal number, nulliparity, and advanced maternal age were each independently associated with the development of pregnancy-related hypertensive conditions. CONCLUSION: Mild and severe pregnancy-related hypertensive disease increases progressively with advancing fetal number from singleton to triplets but is not further increased in quadruplet pregnancies. Multifetal pregnancies should be observed closely for onset of gestational hypertensive disease. LEVEL OF EVIDENCE: II-2. PMID- 16260509 TI - Long-term outcome of uterine artery embolization of leiomyomata. AB - OBJECTIVE: To determine the long-term outcome from uterine artery embolization for leiomyomata. METHODS: In a prospective study, 200 consecutive patients treated with uterine embolization were each followed for 5 years. Outcome, including symptom status compared with baseline, reinterventions, menstrual status, and satisfaction were recorded. Summary statistics were used to report baseline characteristics and outcome at each interval. Predictors of subsequent interventions, failure, and satisfaction with treatment were analyzed using logistic regression and Cox proportional hazards models. Failure was defined as subsequent hysterectomy, definitive myomectomy, repeat embolization, or failure of symptom improvement at the patient's final follow-up interval. RESULTS: Of the 200 patients initially treated, 5-year follow-up was completed in 182 (91%), with 18 patients missing. At 5 years after treatment, 73% had continued symptom control, whereas 36 (20%) had failed or recurred. There had been 25 hysterectomies (13.7%), 8 myomectomies (4.4%), and 3 repeat embolizations (1.6%). Long-term failure was more likely in those not improved at 1 year (relative risk [RR] 5.73; 95% confidence interval [CI] 2.32-14.12, P < .001) and in those with baseline leiomyoma volumes greater than the median (RR 2.18; 95% CI 1.05-4.51, P = .036). After adjustment, patients in the first tertile of leiomyoma volume reduction (< or = 30.5%) were 3 times more likely to be dissatisfied with outcome compared with women in the third tertile (> or = 56.3% volume reduction) (RR 3.23; 95% CI 1 07-9.81, P = .037). CONCLUSION: Uterine embolization provides durable symptom relief for most patients, with a 25% chance of failure of symptom control or recurrence over the course of a 5-year follow-up. LEVEL OF EVIDENCE: II-3. PMID- 16260510 TI - Postmenopausal hormone therapy: does it cause incontinence? AB - OBJECTIVE: To estimate the effect of hormone therapy on risk of stress and urge urinary incontinence. METHODS: The Heart Estrogen/progestin Replacement Study was a randomized, placebo-controlled, double-blinded trial to evaluate daily oral conjugated estrogen (0.625 mg) plus medroxyprogesterone acetate (2.5 mg) therapy for the prevention of heart disease events in women with established heart disease. The 1,208 participants in Heart Estrogen/progestin Replacement Study who reported no loss of urine in the previous 7 days at baseline are included in this analysis. RESULTS: During 4.2 years of treatment, 64% of women randomly assigned to hormone therapy compared with 49% of those assigned to placebo reported weekly incontinence (P < .001). The higher risk of incontinence in the hormone group was evident at 4 months, persisted throughout the treatment period, and was independent of the age of the women. The odds ratios for weekly incontinence among women on hormone therapy compared with placebo were 1.5 for urge incontinence (95% confidence interval [CI] 1.2-1.8; P < .001) and 1.7 for stress incontinence (95% CI 1.5-2.1; P < .001). Four years of treatment with hormone therapy caused an excess risk of 12% for weekly urge incontinence and 16% for weekly stress incontinence; the corresponding numbers needed to harm were 8.6 (95% CI 5.8-16.6) and 6.2 (95% CI 4.6-9.4). CONCLUSION: Estrogen plus progestin therapy increases risk of urge and stress incontinence within 4 months of beginning treatment. LEVEL OF EVIDENCE: I. PMID- 16260511 TI - The effect of ultralow-dose transdermal estradiol on urinary incontinence in postmenopausal women. AB - OBJECTIVE: To estimate the effect of 2 years of treatment with ultralow-dose transdermal estradiol (E2) on incontinence in postmenopausal women. METHODS: Ultra Low Dose Transdermal estRogen Assessment (ULTRA) was a multicenter, randomized, double-blinded, placebo-controlled trial of unopposed ultralow-dose (0.014 mg/d) transdermal E2 for prevention of osteoporosis in 417 postmenopausal women aged 60 to 80 years. Frequency of incontinence episodes was assessed at baseline and after 4 months and 2 years of treatment using a self-reported questionnaire. We used an intention-to-treat analysis to compare change in incontinence frequency, improved (decreased 2 or more episodes per week), unchanged (increased or decreased no more than 1 episode per week), or worsened (increased 2 or more episodes per week) between the E2 and placebo groups among women with and without at least weekly incontinence at baseline. RESULTS: At baseline, the prevalence of at least weekly incontinence was similar between E2 and placebo groups (43%). After 2 years, there was no difference between groups in the proportions of women with incontinence at baseline whose incontinence improved, worsened, or was unchanged. The odds ratio for worsening incontinence in the E2 compared with placebo group was 1.35 (95% confidence interval 0.75 2.42. In women without incontinence at baseline, the odds of developing at least weekly incontinence after 2 years in the E2 compared with placebo group was not significant (odds ratio 1.2, 95% confidence interval 0.7-2.2). CONCLUSION: Two years of treatment with unopposed ultralow-dose transdermal E2 did not substantially change the frequency of incontinence symptoms or alter the risk of developing at least weekly incontinence. LEVEL OF EVIDENCE: I. PMID- 16260513 TI - Birth outcomes in obese women after laparoscopic adjustable gastric banding. AB - OBJECTIVE: This prospective study sought to examine the outcomes of 79 consecutive first pregnancies (> 20 weeks of gestation) in women following laparoscopic adjustable gastric banding (LAGB) for severe obesity. METHODS: The 79 women are from a cohort of 1,382 consecutive patients. The prospectively collected data from 79 first pregnancies has been compared with these patients' previous penultimate pregnancies (n= 40), obstetric histories from matched severely obese subjects (n = 79), and community outcomes. RESULTS: The mean maternal weight gain was 9.6 +/- 9.0 kg, compared with 14.4 +/- 9.7 kg for the 40 penultimate pregnancies of women in this group (P < .001). There was no difference in birth weights: 3,397 g compared with 3,350 g for preband pregnancies, and these were consistent with normal community birth weights. The incidence of pregnancy-induced hypertension (10%) and gestational diabetes (6.3%) were comparable with community levels (12% and 5.5%) and lower than the obese cohort (38% and 19%) and these patients' penultimate pregnancies (45% and 15%). Monitoring and, if necessary, band adjustments during pregnancy provided more favorable maternal weight outcomes (P = .027). Neonatal outcomes, including stillbirths, preterm deliveries, low birth weight, and high birth weight, were consistent with community values. One woman developed anemia during pregnancy. CONCLUSION: Pregnancy outcomes after LAGB are consistent with general community outcomes rather than outcomes from severely obese women. The adjustability of the LAGB assists in achieving these outcomes. Adjustability is appealing because it allows adaptation to the altered requirements of pregnancy. LEVEL OF EVIDENCE: II 2. PMID- 16260512 TI - Remodeling of vaginal connective tissue in patients with prolapse. AB - OBJECTIVE: As pelvic organ prolapse progresses, the morphology of the vagina dramatically changes. The objective of this study was to determine whether these changes observed clinically correlate with histologic and biochemical evidence of tissue remodeling METHODS: After informed consent, full-thickness biopsies of the vaginal apex were obtained at the time of surgery from 77 women. The tissue of 15 premenopausal women with less than stage II prolapse (controls) was compared with that of 62 women with prolapse divided according to their menopausal status. All specimens were examined histologically. Scanning confocal microscopic analysis of fluorescent micrographs was used to quantitate collagen subtypes I, III, and V. Collagen fiber orientation was analyzed by scanning electron microscopy. Gelatin zymography was used to quantitate the expression of the proenzyme and active forms of matrix metalloproteinases (MMP) -2 and -9. Median values were compared using Mann-Whitney U or Kruskal-Wallis tests, where appropriate RESULTS: Vaginal collagen fibers are arranged in a whorled pattern, with collagen III as the predominant fibrillar collagen. The amount of total collagen in the vagina was increased in women with prolapse relative to women without prolapse (P = .054) primarily due to increased expression of collagen III (P = .031). There was no difference in the expression of proMMP-2, active MMP-2, or proMMP-9; however, active MMP-9 was increased in patients with prolapse (P = .030) CONCLUSION: The increase in collagen III and active MMP-9 expression in the vaginal tissues of patients with prolapse suggests that this tissue is actively remodeling under the biomechanical stresses associated with prolapse. LEVEL OF EVIDENCE: II-2. PMID- 16260514 TI - Transient hypothyroxinemia in severe hypertensive disorders of pregnancy. AB - OBJECTIVE: Assess whether and to what extent thyroid function is affected in pregnant women with early and severe hypertensive disorders and in their newborns. METHODS: Patients were 80 women with preeclampsia, hemolysis, elevated liver enzymes, and low platelet count syndrome or gestational hypertension combined with fetal growth restriction in the 24th to 34th week of singleton pregnancies. Maternal thyroid hormone levels and thyroid peroxidase antibodies were determined at admission and 3 months postpartum. Neonatal levels were determined from cord blood at delivery. Maternal hypothyroxinemia was defined as free T(4) (fT(4)) value below 9 pM. RESULTS: At admission 26 (33%) women in the study group had fT(4) levels below 9 pM, with spontaneous normalization during pregnancy. There were no statistically significant differences between thyroid hormone values in women in the study group and 10 normotensive pregnant women in their third trimester. Three months postpartum, 97.5% of patients had normal thyroid hormone levels. Thyroid peroxidase antibodies were elevated in 10% of women postpartum. Their infants, born at a median gestational age of 30 6/7 weeks, had lower cord blood fT(4) and thyroid-stimulating hormone values compared with preterm infants of the comparison group, appropriate for gestational age. Cord blood fT(4) had no correlation with gestational age or maternal fT(4), but there was a significant correlation of cord blood fT(4) with umbilical artery pH. CONCLUSION: Women with severe hypertensive disorders of pregnancy may have transiently lower fT(4) levels, without evidence of a thyroid disorder. Their neonates have lower fT(4) levels at birth unrelated to maternal fT(4), but related to prenatal acidosis. LEVEL OF EVIDENCE: II-2. PMID- 16260515 TI - Fetal fibronectin as a predictor of vaginal birth in nulliparas undergoing preinduction cervical ripening. AB - OBJECTIVE: We sought to evaluate whether the presence of a positive fetal fibronectin (> or = 50 ng/mL) in nulliparous women undergoing preinduction cervical ripening with the intracervical Foley catheter predicted vaginal birth. METHODS: This was a prospective blinded observational trial of nulliparous women undergoing preinduction cervical ripening. We excluded women who had a contraindication to vaginal birth. Cervical and vaginal fetal fibronectin specimens were obtained before preinduction cervical ripening with an intracervical Foley catheter. The managing obstetrician was blinded to these results. RESULTS: A total of 241 women met the inclusion criteria, of which 54.4% delivered vaginally. There was no difference in the rate of vaginal delivery among women with either a positive cervical fetal fibronectin (positive fetal fibronectin 55.8% compared with negative fetal fibronectin 53.3%, P = .70) or positive vaginal fetal fibronectin (positive fetal fibronectin 57.6% compared with negative fetal fibronectin 53.3%, P = .56). Women with a positive cervical fetal fibronectin did have a shorter duration of cervical ripening (fetal fibronectin-positive 229 +/- 220 minutes compared with fetal fibronectin-negative 379 +/- 193 minutes, P < .05), duration of oxytocin (fetal fibronectin-positive 655 +/- 555 minutes compared with fetal fibronectin-negative 731.5 +/- 342 minutes, P < .025) and required lower maximal doses of oxytocin (fetal fibronectin-positive 18.4 mIU/min compared with fetal fibronectin-negative 21.8 mIU/min, P = .005). Women with a positive vaginal fetal fibronectin demonstrated only a shorter duration of cervical ripening compared with their fetal fibronectin negative counterparts (fetal fibronectin-positive 300 +/- 216 minutes compared with fetal fibronectin-negative 345 +/- 201 minutes, P < .05). CONCLUSION: Fetal fibronectin does not predict vaginal delivery in nulliparous women requiring preinduction cervical ripening. LEVEL OF EVIDENCE: II-2. PMID- 16260516 TI - Maternal smoking and its association with birth weight. AB - OBJECTIVE: Maternal smoking has been associated with a reduction in newborn birth weight. We sought to estimate how the pattern of maternal smoking throughout pregnancy influences newborn size. METHODS: One hundred sixty pregnant smoking women were enrolled in a prospective study. We collected data on maternal age, education, prepregnancy body mass index, and parity, as well as alcohol and illicit drug use. Cigarette use was defined as self-reported consumption before pregnancy, at the time of study enrollment, and in the third trimester. Statistical analyses were performed based on bivariate correlations and multiple linear regression. RESULTS: Of the smoking parameters examined, maternal third trimester cigarette consumption was the strongest predictor of birth weight percentile (partial r = -0.23, P < .001). For each additional cigarette per day that a participant smoked in the third trimester, there was an estimated 27 g reduction in birth weight. Prepregnancy smoking volume was not significantly associated with birth weight percentile in bivariate (r = -0.06, P = .47) or multivariable analyses. Additional factors contributing to birth weight include gestational age (partial r = 0.69, P < .001), maternal body mass index (partial r = 0.23, P < .001), and parity (partial r = 0.16, P < .004). In total, these 4 variables explain 61% of the variance in newborn birth weight. CONCLUSION: Maternal third-trimester cigarette consumption is a strong and independent predictor of birth weight percentile. This supports the hypothesis that reductions in maternal cigarette consumption during pregnancy will result in improved birth weight, regardless of the prepregnancy consumption levels. LEVEL OF EVIDENCE: III. PMID- 16260517 TI - Risk factors for spontaneous abortion in early symptomatic first-trimester pregnancies. AB - OBJECTIVE: To evaluate the association of an ultimate diagnosis of miscarriage with various clinical symptoms and historical factors in a cohort of women presenting with pain, bleeding, or both in the first trimester of pregnancy. METHODS: This was a case-control study from a population of women presenting for care with pelvic pain or vaginal bleeding in the first trimester of pregnancy whose diagnoses were not definite upon initial evaluation. Analyses were performed in 2 ways. In one instance cases were defined as women ultimately definitively diagnosed with a miscarriage and controls were defined as women with a pregnancy that did not result in miscarriage (ectopic pregnancy or ongoing intrauterine pregnancy). The second analysis compared women with a miscarriage only to women who had an ongoing intrauterine pregnancy. RESULTS: A total of 2,026 women were evaluated, with 1,192 ultimately diagnosed with a spontaneous abortion, 367 with ectopic pregnancy, and 467 with a viable intrauterine pregnancy. Although many risk factors were individually associated with miscarriage in preliminary analysis, in the final analysis only extremes in age (< 25 and > 35) and the complaint of bleeding (odds ratio [OR] 7.35, 95% confidence interval[CI] 5.74-9.41) were associated with miscarriage. The complaint of pain (OR 0.72, 95% CI 0.57-0.92), human chorionic gonadotropin (hCG) value greater than 500 (hCG < or = 500 IU/mL compared with hCG 501-2000: OR 0.52, 95% CI 0.39-0.69) and concurrent cervical infection (OR 0.69, 95% CI 0.55-0.88) were negatively associated with miscarriage. CONCLUSION: Few factors predict miscarriage in women who present with a symptomatic first trimester pregnancy of unknown location. Heavy bleeding was most strongly associated with miscarriage. Concurrent cervical infections should not be overlooked as a cause of bleeding in early pregnancy and were not associated with miscarriage. LEVEL OF EVIDENCE: II 2. PMID- 16260518 TI - Bladder perforation during tension-free vaginal tape procedures: analysis of learning curve and risk factors. AB - OBJECTIVE: To estimate whether rates of bladder perforation decrease with increasing surgical experience. METHODS: We performed a review of all patients undergoing a tension-free vaginal tape procedure performed by senior resident physicians under the guidance of a single surgeon. Physician experience was assessed by sequentially assigning case numbers to each procedure for each resident. For analysis of learning curve, cases were grouped in fives (ie, first five representing cases 1 to 5, second five cases 6 to 10). RESULTS: Twenty-three residents performed 278 procedures. The median number of cases performed was 13 (range 3 - 22); mean number was 12.1 (sd = +/- 5.6). The rate of perforation was 34.2% (95/278, 95% confidence interval 28.8-39.9%). Age and weight were significantly associated with perforation. The cystotomy group was, on average 4.5 years younger (P = .007) and 7.86 kg (17.3 lb) lighter (P < .001). Rate of injury in the first five series was 40.9%, 30.7% in second series of five, and 25.9% in the third series of five and was statistically significant (linear-by linear association chi(2) = 4.286, df = 1, P = .038). The relationship between the incidence of cystotomy and the cumulative number of cases performed was inversely correlated. As the number of cases a resident completed increased, there was a slight tendency for cystotomy to decrease (P.033). On cystoscopic examination, residents missed 35 of the 95 injuries (37%, 95% confidence interval 27.8-46.9%). CONCLUSION: A learning curve exists for tension-free vaginal tape procedures. Many injuries are missed on initial resident cystoscopic inspection, highlighting the need for comprehensive cystoscopic training during residency. LEVEL OF EVIDENCE: II-3. PMID- 16260519 TI - Involvement of the female genital tract in pemphigus vulgaris. AB - BACKGROUND: Involvement of the genital tract in women with pemphigus vulgaris has rarely been reported. We report 34 patients with pemphigus vulgaris who have involvement of the vulva, vagina, or both. METHODS: We reviewed the clinical data on 34 women treated at a tertiary dermatology referral center from July 1988 to June 2005. RESULTS: Of 34 patients, 21 had labial involvement, 3 had vaginal involvement, and 10 had both labial and vaginal involvement. The mean age of onset of pemphigus vulgaris was 49 years. All patients had pemphigus vulgaris involvement at multiple other sites, and all had other mucous membrane involvement, most commonly oral (97%). Recurrent episodes of genital pemphigus vulgaris were noted in 35%, with a mean of 6 recurrences (range 1-11). In all the patients the pemphigus vulgaris was controlled with systemic and local therapy. Long-term follow-up for a mean of 76 months (range 9-204) indicated that no long term sequela occurred due to genital involvement. CONCLUSION: Involvement of the female genital tract in pemphigus vulgaris is rare. Vulvar lesions occur more commonly than vaginal lesions. With appropriate topical and systemic therapy, patients can have full recovery with no sequelae. LEVEL OF EVIDENCE: II-3. PMID- 16260520 TI - Adherent biofilms in bacterial vaginosis. AB - OBJECTIVE: Bacterial vaginosis is a common infectious disorder. Although known since ancient times, little progress has occurred in identifying causal factors. Our aims were to study the bacterial community structure and the spatial organization of microbiota on the epithelial surfaces of vaginal biopsy specimens. METHODS: We investigated the composition and spatial organization of bacteria associated with the vaginal epithelium in biopsy specimens from 20 patients with bacterial vaginosis and 40 normal premenopausal and postmenopausal controls using a broad range of fluorescent bacterial group-specific rRNA targeted oligonucleotide probes. RESULTS: Bacterial vaginosis was associated with greater occurrence and higher concentrations of a variety of bacterial groups. However, only Gardnerella vaginalis developed a characteristic adherent biofilm that was specific for bacterial vaginosis. CONCLUSION: A biofilm comprised of confluent G vaginalis with other bacterial groups incorporated in the adherent layer is a prominent feature of bacterial vaginosis. LEVEL OF EVIDENCE: II-2. PMID- 16260521 TI - Ten estrogen-related polymorphisms and endometriosis: a study of multiple gene gene interactions. AB - OBJECTIVE: Genetic as well as hormonal factors are known to influence the development and clinical course of endometriosis. We aimed to investigate the association among 10 single nucleotide polymorphisms (SNPs) involved in the estrogen metabolism and endometriosis and to develop a multiple genetic model. METHODS: In a case-control study, we investigated the genotype frequencies of 10 estrogen metabolizing SNPs in 32 patients with endometriosis and 790 healthy controls using sequencing-on-chip-technology with solid-phase polymerase chain reaction on oligonucleotide microarrays: catechol-O-methyltransferase, Val158Met G->A, 17-beta-hydroxysteroid dehydrogenase type 1 (HSD17), vlV A->C, cytochrome P450 (CYP), 17 A2 allele T->C, CYP1A1 MspI RFLP T->C, CYP1A1 Ile462Val A->G, CYP19 Arg264Cys C->T, CYP19 C1558T C->T, CYP 1B1 Leu432Val, CYP1B1 Asn453Ser, and estrogen receptor alpha IVS1 -401>C. Associations and 2-way interaction models between SNPs were calculated by stepwise logistic regression models. RESULTS: In a univariate model, HSD17 vlV A->C was associated with a significantly increased risk of endometriosis (P = .004; odds ratio 3.9, 95% confidence interval 1.6 9.8). When all 2-way interactions of investigated SNPs were ascertained, no significant interactions among SNPs were observed. In a multivariate model, HSD17 vlV A->C was also significantly associated with endometriosis (P = .002). CONCLUSION: We present data on multiple SNPs in patients with endometriosis indicating an association between HSD17 gene variation and the disease. Although not able to demonstrate interaction models of SNPs, we provide evidence of HSD17 vlV A->C as a low penetrance genetic marker of endometriosis. LEVEL OF EVIDENCE: II-2. PMID- 16260523 TI - Assisted reproductive technology and pregnancy outcome. AB - OBJECTIVE: To determine whether the use of assisted reproductive technology (ART) is associated with an increase in chromosomal abnormalities, fetal malformations, or adverse pregnancy outcomes. METHODS: A prospective database from a large multicenter investigation of singleton pregnancies, the First And Second Trimester Evaluation of Risk trial, was examined. Subjects were divided into 3 groups: no ART use, use of ovulation induction (with or without intrauterine insemination), and use of in vitro fertilization (IVF). Multivariate logistic regression analysis was used to assess association between ART and adverse pregnancy outcomes (significance of differences was accepted at P < .05). RESULTS: A total of 36,062 pregnancies were analyzed: 34,286 (95.1%) were spontaneously conceived, 1,222 (3.4%) used ovulation induction, and 554 (1.5%) used IVF. There was no association between ART and fetal growth restriction, aneuploidy, or fetal anomalies after adjustment for age, race, marital status, years of education, prior preterm delivery, prior fetal anomaly, body mass index, smoking history, and bleeding in the current pregnancy. Ovulation induction was associated with a statistically significant increase in placental abruption, fetal loss after 24 weeks, and gestational diabetes after adjustment. Use of IVF was associated with a statistically significant increase in preeclampsia, gestational hypertension, placental abruption, placenta previa, and risk of cesarean delivery. CONCLUSION: Patients who undergo IVF are at increased risk for several adverse pregnancy outcomes. Although many of these risks are not seen in patients undergoing ovulation induction, several adverse pregnancy outcomes are still increased in this group. There was no increased incidence of fetal chromosomal or structural abnormalities in the women who used any type of ART compared with the women who conceived spontaneously. LEVEL OF EVIDENCE: II-2. PMID- 16260522 TI - Third-trimester maternal toxicity with nevirapine use in pregnancy. AB - OBJECTIVE: Nevirapine-based therapy is associated with increased frequency of adverse events among women with CD4+ cell count of 250 cells/microL or greater. We evaluated the safety of nevirapine-based antiretroviral therapy in human immunodeficiency virus (HIV)-1-infected pregnant women. METHODS: We retrospectively evaluated 23 pregnancies managed with nevirapine-based regimens from July 2001 to April 2005. The incidence of adverse events was determined and analyzed by CD4+ cell count of either less than or greater than or equal to 250 cells/microL, and gestational age when nevirapine was initiated. Liver function abnormality was graded according to the National Institute of Allergy and Infectious Diseases Division of AIDS toxicity guidelines. RESULTS: Five of 23 patients (21.7%) started nevirapine-based therapy after 27 weeks of gestation. All 3 cases of adverse events occurred in this group within 6 weeks of initiating therapy and with CD4+ cell count greater than 250 cells/microL. A significant difference was noted in the proportion of patients who developed toxicity while starting nevirapine in the third trimester (3/5, 60%; 95% confidence interval 14.66-94.73) compared with those starting nevirapine earlier in pregnancy (0/18, 0%; 95% confidence interval 0.0-18.53; P < .006). Two patients developed rash, eosinophilia, and liver function abnormality, with one developing clinical hepatitis and renal failure. A third patient had abnormal elevation of liver enzymes but was asymptomatic. CONCLUSION: The incidence of adverse events with nevirapine may be lower than previously reported (13% versus 29%) and may be primarily noted with initiating the drug late in pregnancy. Further study of nevirapine in larger cohorts of HIV-infected pregnant women is warranted to determine the relationship between nevirapine hepatotoxicity and trimester use. LEVEL OF EVIDENCE: II-3. PMID- 16260524 TI - Severe asthma exacerbations during pregnancy. AB - OBJECTIVE: To estimate the frequency of severe asthma exacerbations in pregnant women and to estimate whether there is an association with adverse perinatal outcomes. METHODS: Asthma exacerbations were evaluated in 146 women who were enrolled in a prospective cohort study of asthma and pregnancy. A severe exacerbation was defined as a hospital admission, emergency department presentation, or unscheduled doctor visit for asthma or a course of oral corticosteroids. Women were classified as having mild (n = 63), moderate (n = 34), or severe (n = 49) asthma. RESULTS: Severe exacerbations occurred in 8% (95% confidence interval [CI] 1.3-14.6%) of women with mild asthma, 47% (95% CI 30.3 63.8%) of women with moderate asthma, and 65% (95% CI 52-78.6%) of women with severe asthma at a mean gestational age of 25.1 +/- 0.9 (range 9-39) weeks of gestation. Among women who had severe exacerbations, there were 2 male stillbirths (P = .102) and a significantly increased rate of male low birth weight (P = .03). Maternal age, lung function, body mass index, gravidity, and parity were not different between women who did or those who did not have a severe exacerbation. Maternal pregnancy weight gain was significantly lower in women who had a severe exacerbation (P = .039). Forty-three percent of severe exacerbations occurred in winter, 34% were associated with self-reported viral infection, and 29% with nonadherence to inhaled corticosteroid medication. CONCLUSION: The exacerbation rate among pregnant women with asthma is high and associated with poor outcomes for the male fetus. Improvements in asthma management to prevent severe exacerbations may lead to a better outcome for both mother and baby. LEVEL OF EVIDENCE: II-2. PMID- 16260525 TI - Ethical issues related to medical expert testimony. AB - The obstetrician-gynecologist who provides expert witness testimony is recognized as an important participant in the medical liability system. He or she must define a standard of care and opine whether the standard has been breached and whether any perceived injury was caused by the breach. The American College of Obstetricians and Gynecologists (ACOG) ethical guidelines insist that the testimony be objective (free from intentioned distortion) and that the witness not assume an advocacy or partisan role. The ethical and professional boundaries of appropriate expert testimony as delineated by ACOG are more restrictive than the legal boundaries. Members of ACOG should be held to more restrictive ACOG guidelines and egregious testimony condemned by ACOG. Prospective peer review, increased judicial review, and testimony banks are other examples of methods to improve the quality of expert witness testimony. Alternatives to litigation for medical liability disputes and further tort reform might also make the system fairer and more sustainable. PMID- 16260526 TI - Recognition and prevention of fetal alcohol syndrome. AB - Alcohol use among women of childbearing age is prevalent in the United States, with approximately 1 in 5 nonpregnant women reporting binge drinking (5 or more drinks on any one occasion) and 1 in 25 pregnant women reporting binge drinking. Alcohol use during pregnancy results in a spectrum of adverse outcomes known as fetal alcohol spectrum disorders. Fetal alcohol syndrome (FAS) is one of these disorders. Fetal alcohol syndrome is characterized by specific facial abnormalities and significant impairments in neurodevelopment and physical growth. Early identification of children with FAS has been shown to enhance their long-term outcomes. In an effort to improve clinical recognition of children with this condition, Centers for Disease Control and Prevention (CDC) was directed by Congress in 2002 to lead the development of uniform diagnostic criteria for FAS and other prenatal alcohol-related conditions. The purpose of this commentary is to provide clinicians a summary of the report released by CDC describing the current diagnostic criteria for FAS. In addition, advancements have been made in screening and brief interventions for alcohol use disorders in women who have the potential to make significant strides in the prevention of FAS spectrum disorders. Knowledge of the diagnostic criteria for FAS can lead to increased identification of the syndrome in infants and children and the provision of appropriate medical and support services. Screening for and intervening with women at risk for an alcohol-exposed pregnancy can prevent FAS and other fetal alcohol spectrum disorders. PMID- 16260527 TI - Why can't a man be more like a woman? Sex, power, and politics. AB - Conflicts between male and female reproductive agendas continue to play out in contemporary issues of sex, power, and politics. Viewing gender through the lens of biologic evolutionary psychology reveals persistent controversies surrounding women's rights. The history of oral contraceptives compared with that of erectile dysfunction drugs is just one example of the disparity between female and male reproductive choices. Contraceptives, maternal mortality, abortion, and domestic violence are issues directly influenced by politics, religion, and gender biases. Ultimately, everything that can we can do to give women control over their bodies and their fertility is not only just and humane, but it also changes the world for the better. The United States must restore its leadership in international family planning--ensuring reproductive freedoms could be the genesis of other freedoms. PMID- 16260528 TI - Perinatal depression: a systematic review of prevalence and incidence. AB - OBJECTIVE: We systematically review evidence on the prevalence and incidence of perinatal depression and compare these rates with those of depression in women at non-childbearing times. DATA SOURCES: We searched MEDLINE, CINAHL, PsycINFO, and Sociofile for English-language articles published from 1980 through March 2004, conducted hand searches of bibliographies, and consulted with experts. METHODS OF STUDY SELECTION: We included cross-sectional, cohort, and case-control studies from developed countries that assessed women for depression during pregnancy or the first year postpartum with a structured clinical interview. TABULATION, INTEGRATION, AND RESULTS: Of the 109 articles reviewed, 28 met our inclusion criteria. For major and minor depression (major depression alone), the combined point prevalence estimates from meta-analyses ranged from 6.5% to 12.9% (1.0 5.6%) at different trimesters of pregnancy and months in the first postpartum year. The combined period prevalence shows that as many as 19.2% (7.1%) of women have a depressive episode (major depressive episode) during the first 3 months postpartum; most of these episodes have onset following delivery. All estimates have wide 95% confidence intervals, showing significant uncertainty in their true levels. No conclusions could be made regarding the relative incidence of depression among pregnant and postpartum women compared with women at non childbearing times. CONCLUSION: To better delineate periods of peak prevalence and incidence for perinatal depression and identify high risk subpopulations, we need studies with larger and more representative samples. PMID- 16260529 TI - Urinary tract infection in women. AB - Urinary tract bacterial infections are common in women. Moreover, they tend to recur throughout life and in the same relatively small group of women. In most cases, bladder and renal infections are asymptomatic and manifest by demonstrating coincidental bacteriuria. In some instances, however, especially with frequent sexual activity, pregnancy, stone disease, or diabetes, symptomatic cystitis or pyelonephritis develops and antimicrobial therapy is indicated. In most cases, cystitis is easily managed with minimal morbidity. When acute pyelonephritis develops in an otherwise healthy woman, however, consideration for ureteral obstruction is entertained. If her clinical response to proper therapy is not optimal, then imaging studies are indicated. Pregnancy is a common cause of obstructive uropathy, and severe renal infections are relatively common. Because they usually arise from preexisting covert bacteriuria, experts recommend screening and eradication of these silent infections as a routine prenatal practice. PMID- 16260530 TI - Lift up your eyes. PMID- 16260535 TI - Ovarian conservation at the time of hysterectomy for benign disease. PMID- 16260537 TI - Simpson, Semmelweis, and transformational change. PMID- 16260536 TI - Ovarian conservation at the time of hysterectomy for benign disease. PMID- 16260538 TI - Diagnosis of anal sphincter tears to prevent fecal incontinence: a randomized controlled trial. PMID- 16260539 TI - Temporary Erb-Duchenne palsy without shoulder dystocia or traction to the fetal head. PMID- 16260540 TI - Endometrial effects of bazedoxifene acetate, a novel selective estrogen receptor modulator, in postmenopausal women. PMID- 16260542 TI - Efficacy and safety of isopropanolic black cohosh extract for climacteric symptoms. PMID- 16260543 TI - ACOG Practice Bulletin #68: Antiphospholipid syndrome. PMID- 16260544 TI - ACOG Committee Opinion #320: Partnering with patients to improve safety. PMID- 16260545 TI - ACOG Committee Opinion #321: Maternal decision making, ethics, and the law. PMID- 16260546 TI - ACOG Committee Opinion #322: Compounded bioidentical hormones. PMID- 16260547 TI - ACOG Committee Opinion #323: Elective coincidental appendectomy. PMID- 16260548 TI - ACOG Committee Opinion #324: Perinatal risks associated with assisted reproductive technology. PMID- 16260549 TI - Case reports: catalysts for improving patient care. PMID- 16260550 TI - Uterine anomalies, induction of labor, and uterine rupture. PMID- 16260551 TI - Postcesarean pulmonary embolism, sustained cardiopulmonary resuscitation, embolectomy, and near-death experience. AB - BACKGROUND: Survival after surgical embolectomy for massive postcesarean pulmonary embolism causing sustained cardiac arrest is rare. CASE: One day after an uneventful cesarean delivery, a woman developed cardiac asystole and apnea due to pulmonary embolism. Femoral-femoral cardiopulmonary bypass performed during continuous cardiopulmonary resuscitation allowed a successful embolectomy. Upon awakening, the patient reported a near-death experience. Pulmonary embolism causes approximately 2 deaths per 100,000 live births per year in the United States, and postcesarean pulmonary embolism is probably more common than pulmonary embolism after vaginal delivery. CONCLUSION: Massive pulmonary embolism is a potentially treatable catastrophic event after cesarean delivery, even if continuous cardiopulmonary resuscitation is required until life-saving embolectomy is done. PMID- 16260552 TI - Massive pulmonary embolism in pregnancy treated with tissue plasminogen activator. AB - BACKGROUND: Systemic thrombolysis with tissue plasminogen activator (t-PA) in pregnancy is still considered an experimental treatment. Several reports have described the successful use of t-PA in the setting of hemodynamic instability in gravidas with massive pulmonary emboli. CASE: A 34-year-old woman received a diagnosis of severe pulmonary embolism at 23 weeks of gestation. She developed pulmonary hypertension and became hemodynamically unstable. Thrombolytic therapy using t-PA was administered. The patient tolerated thrombolysis well and delivered at term. No placental abnormalities were identified on ultrasonogram or after delivery. The patient was also found to be a heterozygous carrier of prothrombin G20210A mutation. CONCLUSION: We describe the successful thrombolysis with t-PA of a massive, life-threatening pulmonary embolism without complications followed by a term delivery. PMID- 16260553 TI - Massive pulmonary embolism in pregnancy treated with catheter fragmentation and local thrombolysis. AB - BACKGROUND: Catheter-directed thromboembolus fragmentation and thrombolysis is used with success for treatment of pulmonary embolism with hemodynamic decompensation in nonpregnant patients, but information on its use during pregnancy is limited. We report successful treatment of massive bilateral pulmonary emboli in the third trimester of pregnancy. CASE: A 29-year-old multigravida at 30 weeks of gestation presented with dyspnea, chest pain, heart palpitations, and syncope. A computed tomographic angiogram demonstrated massive bilateral central pulmonary emboli. Despite heparin and oxygen therapy, aggressive fluid resuscitation and pressor treatment, hypotension persisted, and there were prolonged, deep fetal heart rate decelerations. Emergency percutaneous pulmonary artery catheter thrombus fragmentation, followed by local infusion of tissue plasminogen activator, was performed. The patient recovered rapidly and was discharged from the hospital on subcutaneous low-molecular-weight heparin. She was delivered of a normal, healthy infant at term. CONCLUSION: Catheter directed mechanical fragmentation and local thrombolytic infusion therapy is a treatment option for pulmonary embolism with hemodynamic decompensation in pregnancy. Advantages are rapid clot lysis with consequent return of normal hemodynamics and uterine perfusion and avoidance of systemic thrombolytics and associated risk of bleeding complications. PMID- 16260554 TI - Second-trimester rudimentary uterine horn pregnancy: rupture after labor induction with misoprostol. AB - BACKGROUND: Uterine anomalies are often first suspected after bimanual or ultrasonographic examination. Currently there are no specific recommendations for further evaluation of asymptomatic women with suspected uterine anomalies in pregnancy. CASE: A young primigravida with a history of an ultrasound diagnosis of bicornuate uterus presented with mild abdominal pain. An ultrasound examination showed a viable 18-week fetus with anhydramnios in the left uterine horn. Labor induction with misoprostol culminated in uterine rupture. At laparotomy, a ruptured left noncommunicating rudimentary uterine horn of a unicornuate uterus was noted. CONCLUSION: Pregnancies within noncommunicating uterine horns significantly increase the risk of potentially catastrophic outcome, therefore, consideration should be given to performing 3-dimensonal ultrasonography and/or magnetic resonance imaging examinations to determine the nature of uterine anomalies. Caution should be exercised if prostaglandins are considered for use in this setting. PMID- 16260555 TI - Percutaneous transluminal coronary angioplasty and stent placement in pregnancy. AB - BACKGROUND: Myocardial infarction (MI) is uncommon during pregnancy. As the average maternal age increases and assisted reproductive technology allows for very advanced maternal ages, so too may the incidence of MI during pregnancy. Percutaneous transluminal coronary angioplasty (PTCA) with stent placement is an attractive option for treatment of MI in pregnancy when revascularization is required. CASE: We present a gravida with an ST elevation MI during the third trimester, who was treated with emergent PTCA, stent placement, and platelet inhibitors, and we discuss the patient's subsequent obstetric and anesthetic management. CONCLUSION: Percutaneous transluminal coronary angioplasty with stent placement may be used during the third trimester with successful outcome. PMID- 16260557 TI - Splenosis mimicking pelvic mass. AB - BACKGROUND: Splenosis is a rare complication after splenectomy and generally does not cause clinical symptoms. CASE: A 34-year-old woman who had splenectomy in childhood after trauma presented with a pelvic mass. Ultrasound examination demonstrated a right adnexal mass and a myomatous uterus. Exploratory laparotomy revealed multiple splenic implants along the small and large bowel and in the ileocecal region, including the appendix. Total hysterectomy, right salphengectomy, and appendectomy were performed. CONCLUSION: After splenectomy, splenic implants may mimic benign or malignant tumors of the pelvis and may require surgical exploration. PMID- 16260556 TI - Management of interstitial pregnancy using selective uterine artery embolization. AB - BACKGROUND: Interstitial pregnancy is a rare and dangerous form of ectopic pregnancy which is treated by surgery, medical treatment, or both. Management options are not standardized. Currently, conservative nonsurgical treatment seems to be an alternative method in case of interstitial pregnancy. CASE: A right interstitial pregnancy was diagnosed in a 28-year-old woman. She was successfully treated by 2 courses of systemic methotrexate (1 mg/kg) 24 hours apart followed by selective uterine artery embolization. The postembolization course was uneventful, and no rupture occurred. Ten weeks after embolization, human chorionic gonadotropin level was negative. CONCLUSION: Uterine embolization associated with methotrexate can be used successfully in treating selected cases of early interstitial pregnancy. We hypothesize that this procedure combined with methotrexate could reduce hemorrhagic risk. PMID- 16260558 TI - Laparoscopic management of abdominal pelvic splenosis. AB - BACKGROUND: Splenosis is an unusual cause of pelvic pain. CASE: A 37-year-old nulligravida presented with a 10-year history of chronic pelvic and abdominal pain that was diagnosed as splenosis via laparoscopy. The patient was previously advised against surgical intervention. We report our experience with laparoscopic excision of the above splenosis. CONCLUSION: Splenosis should be considered in patients with a history of splenic injury and abdominal pain. Laparoscopic excision of splenic nodules with the argon beam coagulator is a well-suited method of treatment for symptomatic splenosis. PMID- 16260559 TI - Iatrogenic fetal injury. AB - BACKGROUND: Iatrogenic fetal injury during cesarean delivery is a serious but underreported complication. CASE: A distal iatrogenic amputation of a digit occurred during a cesarean delivery. CONCLUSION: Obstetricians should be aware of this potential complication. PMID- 16260560 TI - Endometrial carcinoma after endometrial resection for dysfunctional uterine bleeding. AB - BACKGROUND: Endometrial destruction is an accepted conservative surgical approach for women with dysfunctional uterine bleeding. However, this procedure cannot guarantee complete removal of the entire endometrium. The possibility exists that endometrial carcinoma may develop even years after such procedure. CASE: We report on a case of endometrial carcinoma, which was diagnosed 3 years after hysteroscopic resection of the endometrium for dysfunctional uterine bleeding in a patient with no risk factors. CONCLUSION: Endometrial carcinoma after hysteroscopic endometrial ablation is still a possibility even when strict selection criteria are applied. PMID- 16260561 TI - Delayed leiomyoma degeneration after microwave endometrial ablation. AB - BACKGROUND: Microwave endometrial ablation is an effective treatment for dysfunctional uterine bleeding. Patients with leiomyomata, including submucosal leiomyomata up to 3 cm, may also be treated with microwave endometrial ablation. CASES: A 46-year-old woman with multiple leiomyomata and menometrorrhagia underwent microwave endometrial ablation. Two months after microwave endometrial ablation, she developed signs of peritoneal irritation. A negative laparoscopy excluded a thermal bowel injury. Imaging and clinical examination ultimately determined that her symptoms were due to leiomyoma degeneration. A 38-year-old woman with menometrorrhagia and leiomyomata underwent microwave endometrial ablation. Fifteen days after microwave endometrial ablation, she developed signs of peritoneal irritation. With a presumptive clinical diagnosis of microwave endometrial ablation degeneration, the patient was expectantly managed with pain medications and observation. CONCLUSION: Fibroid degeneration may have a delayed presentation after microwave endometrial ablation. Thermal bowel injury must be excluded in a patient presenting with signs of peritoneal irritation after microwave ablation of the endometrium before diagnosing leiomyoma degeneration, which can be managed expectantly. PMID- 16260562 TI - Twin pregnancy complicated by severe hemolytic disease of the fetus and newborn due to anti-g and anti-C. AB - BACKGROUND: Hemolytic disease of the fetus and newborn caused by anti-G antibodies is rare, and in most previously reported cases, leads to a mild anemia. The RhG antigen is usually found in association with both RhD and RhC. We report a case of a twin pregnancy affected by both anti-G and anti-C alloantibodies leading to severe hemolytic disease of the fetus and newborn requiring multiple intrauterine transfusions and prolonged postnatal therapy. CASE: A patient with a prolonged history of previously affected pregnancies due to anti-D and anti-C was subsequently found to be affected with anti-G instead. She required aggressive therapy during her pregnancy, initially with intravenous immune globulin and plasmapheresis until umbilical blood sampling and intrauterine transfusions were feasible. CONCLUSION: Although hemolytic disease of the fetus and newborn due to anti-G antibodies is rare and usually mild, these pregnancies should be followed up closely and in utero therapy should be offered if necessary. PMID- 16260563 TI - Severe hemolytic disease of the newborn due to anti-Cw. AB - BACKGROUND: Pregnancies complicated by Rh isoimmunization have decreased significantly since the widespread use of Rh immune globulin. Uncommon red blood cell antigens have therefore become more clinically evident. We report a case of anti-Cw immunization that resulted in severe fetal anemia that required multiple transfusions. CASE: A 28-year-old multigravida presented to our service at 18 weeks of gestation with her fourth pregnancy. Her pregnancy was complicated by anti-Cw isoimmunization that resulted in severe fetal anemia requiring in utero fetal blood transfusions. CONCLUSION: While previous reports recommend only postpartum surveillance when Cw isoimmunization is present, we report a case resulting in severe fetal anemia. PMID- 16260564 TI - Spontaneous resolution of mirror syndrome. AB - BACKGROUND: Mirror syndrome, a rare condition characterized by maternal anasarca in a pregnancy complicated by fetal hydrops, may have a devastating fetal outcome and significant maternal morbidity. CASE: We report a case of mirror syndrome caused by parvovirus B19 infection, which resolved spontaneously with good fetal and maternal outcome. CONCLUSION: The pathogenesis of mirror syndrome is not understood. The trigger for mirror syndrome may be derived from a compromised fetus or placenta. PMID- 16260565 TI - Preterm torsion of a gravid uterus didelphys horn of a twin pregnancy. AB - BACKGROUND: Simultaneous pregnancy in each horn of a uterus didelphys is a rare and complex clinical situation. CASE: Torsion of one horn of a gravid didelphic uterus during a twin pregnancy was diagnosed during cesarean delivery after erroneous diagnosis of abruptio placentae. CONCLUSION: Localization of the placenta in a pregnancy complicated by uterine didelphys may improve the ability to diagnose hemiuterus torsion. PMID- 16260566 TI - Herpes simplex virus hepatitis causing acute liver dysfunction and thrombocytopenia in pregnancy. AB - BACKGROUND: Herpes simplex virus (HSV) hepatitis in pregnant women is a rare condition. We report a case confirmed by liver biopsy and successfully treated with empiric intravenous acyclovir. CASE: A 25-year-old primigravida at 34 weeks of gestation presented with fever, thrombocytopenia, and markedly elevated liver enzymes. The patient was treated empirically and was delivered by cesarean. After delivery failed to correct her condition, a liver biopsy revealed HSV hepatitis. The fetus was unaffected and the patient recovered with an extended course of acyclovir. CONCLUSION: Pregnant women are susceptible to disseminated HSV causing hepatitis. A high index of suspicion is necessary to diagnose HSV hepatitis and begin appropriate treatment with acyclovir. Herpes simplex virus hepatitis should be included in the differential diagnosis for liver failure during pregnancy. PMID- 16260567 TI - Life-threatening maternal and fetal macrocytic anemia from antiretroviral therapy. AB - BACKGROUND: Antiretroviral therapy is recommended for human immunodeficiency virus (HIV)-infected patients during pregnancy to reduce the vertical transmission to the newborn. Complications from this therapy are uncommon. CASE: A 38-year-old HIV-positive pregnant woman was treated with lamivudine and zidovudine. At 28 weeks of gestation, her hemoglobin had fallen to 4.6 g/dL with an mean corpuscular volume (MCV) of 126 microm. At 36 weeks the fetal biophysical profile was abnormal. A pale hydropic infant was delivered via emergency cesarean, with a hemoglobin of 2.1 gm and MCV of 131 microm. The newborn hemoglobin normalized after withdrawal of the neonatal retroviral therapy. CONCLUSION: Maternal-fetal macrocytic anemia may complicate antiretroviral therapy. PMID- 16260568 TI - Budd-Chiari syndrome, systemic lupus erythematosus, and secondary antiphospholipid antibody syndrome in pregnancy. AB - BACKGROUND: Hypercoaguable states such as the antiphospholipid antibody syndrome may be associated with thromboses in any vascular bed. CASE: This case report demonstrates clinical manifestations of Budd-Chiari syndrome during pregnancy, diagnostic dilemmas, and suggestions for prevention of serious thromboembolic complications. CONCLUSION: Patients with antiphospholipid antibody syndrome and previous thromboses in any vascular bed who are considering pregnancy should be considered candidates for full anticoagulation throughout the entirety of gestation and the puerperium. Anti-Xa levels may reflect inadequate dosing of low molecular weight heparin, particularly during the first trimester, and should be monitored frequently. In patients with suspected hepatic venous thrombosis, Doppler evaluation may be inadequate to establish the diagnosis. PMID- 16260569 TI - Acute onset of polymyalgia rheumatica in pregnancy. AB - BACKGROUND: Polymyalgia rheumatica is uncommon in young women and remains a diagnostic challenge for pregnant women. CASE: A 28-year-old pregnant woman developed polymyalgia rheumatica in the third trimester. Laboratory investigations revealed elevated erythrocyte sedimentation rate and C-reactive protein levels with normal muscle enzyme levels and seronegativity for rheumatoid factor. Although her symptoms deteriorated as pregnancy progressed, she drastically improved by treatment with prednisone. She underwent cesarean delivery at 39 weeks. She was relapse-free of polymyalgia rheumatica after discontinuation of prednisone on the 50th postoperative day. CONCLUSION: The diagnosis of polymyalgia rheumatica is important to properly manage pregnancy. PMID- 16260570 TI - Goodpasture syndrome in a pregnant woman. AB - BACKGROUND: Goodpasture syndrome, an immunologic disorder characterized by glomerulonephritis and pulmonary hemorrhage, rarely presents in pregnancy. CASE: We describe a patient who was diagnosed with Goodpasture syndrome in her second trimester. She required daily hemodialysis, intermittent plasmapheresis, and immunosuppressive therapy. Her pregnancy was complicated by hypertension, and she delivered a low birth weight neonate prematurely at 26 4/7 weeks of gestation by cesarean due to nonreassuring fetal status. Deterioration in the fetal status may have been secondary to complications of hypertension, in addition to prematurity. CONCLUSION: Goodpasture syndrome in pregnancy may be associated with significant maternal and fetal morbidity. PMID- 16260571 TI - Retropubic hematoma after transobturator sling procedure. AB - BACKGROUND: The transobturator tape procedure is one of the newer minimally invasive sling procedures used for the surgical treatment of genuine stress urinary incontinence. CASES: Two cases of retropubic hematomas following transobturator tape procedure are reported. One patient was managed conservatively and did not require reoperation, and the other patient required computed tomography-guided drainage of the hematoma. In both cases the patients' hematomas resolved, and they remained continent 3-6 months after surgery. CONCLUSION: The transobturator tape procedure is a minimally invasive alternative to the tension-free vaginal tape operation for stress urinary incontinence, but it may be associated with vascular complications. PMID- 16260572 TI - Hematometra in a patient with Cornelia De Lange syndrome. AB - BACKGROUND: Hematometra is usually the result of developmental anomalies or may be secondary to cervical obstruction. Abnormal uterine contractile function (atony) would be an uncommon cause of hematometra. CASE: An 18-year-old female with Cornelia De Lange syndrome and abdominal pain was found to have a hematometra on ultrasound examination. On pelvic examination, her cervical canal was patent and was easily dilated, but the hematometra did not drain until suprapubic pressure was applied. Two weeks postoperatively, pelvic magnetic resonance imaging showed a markedly thinned uterine myometrium and a recurrent hematometra, prompting the decision to perform a hysterectomy. CONCLUSION: Hematometra in a patient with Cornelia De Lange syndrome may be the result of abnormal uterine contractile function. PMID- 16260573 TI - Acute coronary syndrome and preeclampsia. AB - BACKGROUND: Myocardial ischemia and infarction are rare during pregnancy. Approximately 150 cases of myocardial infarction during pregnancy have been documented in the literature worldwide (predominantly anterior wall). However, elevated troponin levels have been reported in patients with preeclampsia. CASES: We describe 2 patients with preeclampsia who presented with acute myocardial ischemia; 1 with ST segment elevation and one non-ST segment elevation at EKG. Our patients clearly had acute coronary syndromes with troponin levels much higher than would be accounted for by preeclampsia. CONCLUSIONS: From the existing body of literature regarding this patient population, it is unclear why there is a higher incidence of adverse myocardial events. It may be that coronary ischemia has been missed in patients with preeclampsia. Troponin I is now readily available for detection of myocardial damage and should be used in this patient population only when clinically indicated, such as when chest discomfort or new electrocardiogram changes are observed. Patients with preeclampsia may be at higher risk for coronary events, and troponin I levels could be a valuable tool with which to monitor women who develop related symptoms. PMID- 16260574 TI - Pregnancy outcome in patients with pulmonary arterial hypertension receiving prostacyclin therapy. AB - BACKGROUND: Pregnancy is contraindicated in cases of pulmonary hypertension, a highly morbid disease affecting young women of childbearing age. CASES: We describe the pregnancies of 3 patients with pulmonary arterial hypertension (idiopathic, Eisenmenger syndrome, and related to systemic lupus erythematosus). They received epoprostenol and low-molecular-weight heparin throughout pregnancy. The patient with Eisenmenger syndrome started epoprostenol in gestational week 16. Cesarean delivery under general anesthesia was performed at 28-33 weeks of gestation; early delivery was necessary in the patient with Eisenmenger syndrome because of fetal growth restriction. All deliveries were uneventful, and birth weights were 1,700, 1,500, and 795 g. There were no postpartum complications. CONCLUSION: Pregnancy in women with pulmonary hypertension should still be considered high risk for both mother and child, but stable patients on epoprostenol may successfully complete pregnancy. PMID- 16260575 TI - Burns in pregnancy. AB - BACKGROUND: Treatment of a major burn injury during pregnancy must incorporate modifications in management resulting from gestational physiologic changes. CASE: A 25-year-old woman, at 34 weeks of gestation, sustained a major burn injury at home. She required ventilatory support, invasive hemodynamic monitoring, and massive fluid resuscitation. Labor was augmented and a spontaneous vaginal delivery of a healthy neonate was achieved. Later, wound autografting was performed. CONCLUSION: Pregnancy-induced physiologic changes affect key factors in the management of the burned patient, including airway management and hemodynamic support. Multidisciplinary management is essential to achieve the best possible outcome. PMID- 16260576 TI - Cavernous hemangioma: diffuse enlarged venous spaces within the myometrium in pregnancy. AB - BACKGROUND: Diffuse enlarged vessels throughout the myometrium are very rare. This case illustrates the diagnosis and clinical management of a pregnancy complicated by large vessels diffusely distributed throughout the myometrium. CASE: A primigravida measured large for dates at 27 weeks gestation. Ultrasonography demonstrated tubular echolucent spaces throughout the myometrium. No flow could be detected within them by color or spectral Doppler. During cesarean delivery blood loss was 1,700 mL, but the uterus was successfully closed with 3 suture layers. A biopsy specimen showed myometrium containing large vascular spaces thought to be veins, consistent with a hemangioma. CONCLUSION: Enlarged vascular spaces diffusely distributed throughout the myometrium proved to be a cavernous hemangioma. Cesarean delivery in the present case produced some additional bleeding that was easily controlled, and the uterus was closed without incident. PMID- 16260577 TI - An unusual cause of sepsis during pregnancy: recognizing infection with chlamydophila abortus. AB - BACKGROUND: Chlamydophila abortus (formerly Chlamydia psittaci serovar 1) is a rare but severe cause of gestational septicemia, with particular problems in diagnosis and clinical management. CASE: A 32-year-old woman in her fourth pregnancy (16th week of gestation) presented with progressive septicemia after extensive contact with abortive material from her goat flock. Treatment with levofloxacin could not prevent abortion. Multiorgan failure requiring catecholamines and artificial ventilation developed in the patient. After the agent was identified by polymerase chain reaction from acute-phase serum, macrolides were administered and yielded clinical improvement. The patient fully recovered. There were no sequelae in the subsequent 6 months. CONCLUSION: Cp abortus must be considered in gestational septicemia after contact with ruminants. Polymerase chain reaction from acute-phase serum is a quick and easy way to establish the diagnosis. Macrolide antibiotics are still the treatment of choice. PMID- 16260578 TI - Lichen sclerosus involving the vagina. AB - BACKGROUND: A review of the English literature since 1940 did not reveal a reported case of lichen sclerosus involving the vaginal mucosa. Diagnosis of lichen sclerosus involving the vagina must thus be a rare occurrence. CASE: This report presents the findings on a 54-year-old white woman with a history of lichen sclerosus involving the vulva. She was found to have lichen sclerosus involving the vaginal mucosa extending to the posterior vaginal fornix. The patient was started on the use of topical clobetasol ointment 0.05% to the vulva to be used twice daily for 1 month, at bedtime for 2 months, and every other day for 3 months. At follow-up, the vulvar and vaginal lichen sclerosus was unchanged, but the patient was asymptomatic. She was using the clobetasol 1 to 2 times per week. CONCLUSION: Lichen sclerosus involving the vagina is a rare occurrence. Each case must be assessed separately and therapy initiated accordingly in each circumstance. Biopsy must be performed in all cases to identify the disease process and rule out malignancy. PMID- 16260579 TI - Pasteurella multocida bacteremia and tuboovarian abscess. AB - BACKGROUND: Tuboovarian abscess is commonly associated with sexually transmitted pathogens. We report a tuboovarian abscess caused by a rare pathogen, Pasteurella multocida, which was managed conservatively. CASE: A 50-year-old sexually inactive woman presented with suprapubic pain and fever. Cat scratches were seen on her hand. Ultrasonography showed a 7.9-cm complex cystic adnexal structure. Her fever persisted despite broad-spectrum parenteral antibiotics. After placement of a transvaginal drain, the patient defervesced, and her pain improved. Both blood cultures and cyst aspirates grew Pasteurella multocida. CONCLUSION: Tuboovarian abscess secondary to rare pathogens must be considered in the differential diagnosis of acute febrile pelvic illness in a non-sexually active woman. Minimally invasive drainage procedures may avoid surgery in patients failing initial antibiotic therapy. PMID- 16260580 TI - The effect of NCX4016 [2-acetoxy-benzoate 2-(2-nitroxymethyl)-phenyl ester] on the consequences of ischemia and reperfusion in the streptozotocin diabetic rat. AB - The aim of this study was to assess the effect of chronic administration of NCX4016 [2 acetoxy-benzoate 2-(2-nitroxymethyl)-phenyl ester], a nitric oxide releasing aspirin derivative on the consequences of coronary artery occlusion in streptozotocin-diabetic rats. Rats were made diabetic by injection of streptozotocin (60 mg kg(-1)) and received insulin (2.5 U kg(-1) s.c.) daily for 4 weeks. Animals received vehicle (1 ml kg(-1) polyethylene glycol), aspirin (65.2 mg kg(-1)), NCX4016 (60 mg kg(-1)), or (iv) NCX4016 (120 mg kg(-1)) orally, once daily for the last 5 days before coronary artery occlusion (CAO). One hour after the last dose, pentobarbital-anesthetized rats were subjected to CAO for 30 min followed by 120-min reperfusion. Neither drug significantly modified initial hemodynamics or plasma glucose levels compared with vehicle treatment in either nondiabetic or diabetic rats. Neither drug modified the total ventricular premature beat (VPB) count in normal animals, although NCX4016, but not aspirin, reduced the total VPB count and the incidence of ventricular tachycardia in diabetic rats. In nondiabetic animals, both aspirin and NCX4016 reduced infarct size. However, in diabetic rats, infarct size was reduced only by the larger dose of NCX4016 (120 mg kg(-1)) but not by aspirin or the lower dose of NCX4016. These results demonstrate that the cardioprotective effects of NCX4016 are reduced in the presence of diabetes compared with the effects seen in nondiabetic animals. In summary, the present study confirms the protective effect of NCX4016 against ischemia-reperfusion injury in the normal rat heart and demonstrates for the first time its protective effect in the heart of streptozotocin-diabetic rats. PMID- 16260581 TI - Blood glucose-lowering nuclear receptor agonists only partially normalize hepatic gene expression in db/db mice. AB - Agonists of the nuclear receptors peroxisome proliferator-activated receptor (PPAR) gamma, PPARalpha, and liver X receptors (LXRs) reduce blood glucose in type 2 diabetic patients and comparable mouse models. Since the capacity of these drugs to normalize hepatic gene expression is not known, we compared groups of obese diabetic db/db mice treated with agonists for PPARgamma [rosiglitazone (Rosi); 10 mg/kg/day], PPARalpha [Wy 14643 (Wy; 4-chloro-6-(2,3-xylidino)-2 pyrimidinyl)thioacetic acid); 30 mg/kg/day], and LXR [T0901317 (T09; N-(2,2,2 trifluoroethyl)-N-[4-[2,2,2-trifluoro-1-hydroxy-1(trifluoromethyl)-ethyl]phenyl] benzenesulfonamide); 40 mg/kg/day] and from untreated nondiabetic litter mates (db/+) by oligonucleotide microarrays and quantitative reverse transcriptase polymerase chain reaction. The 10-day treatment period of db/db mice with Rosi, Wy, and T09 altered expression of 300, 620, and 735 genes including agonist specific target genes, respectively. However, from the 337 genes differentially regulated in untreated db/+ versus db/db animals, only 34 (10%), 51 (15%), and 82 (24%) were regulated in the direction of the db/+ group by Rosi, Wy, and T09, respectively. Gene expression normalization by drug treatment involved glucose homeostasis, lipid homeostasis, and local glucocorticoid activation. In addition, our data pointed to hitherto unknown interference of these nuclear receptors with growth hormone receptor gene expression and endoplasmic reticulum stress. However, many diabetes-associated gene alterations remained unaffected or were even aggravated by nuclear receptor agonist treatment. These results suggest that diabetes-induced gene expression is minimally reversed by potent blood glucose lowering nuclear receptor agonists. PMID- 16260582 TI - Therapeutic promise of proteinase-activated receptor-2 antagonism in joint inflammation. AB - Biological therapies such as tumor necrosis factor-alpha inhibitors have advanced the treatment of rheumatoid arthritis, but one-third of patients do not respond to such therapy. Furthermore, these inhibitors are now usually administered in combination with conventional disease-modifying antirheumatic drugs, suggesting they have not achieved their early promise. This study investigates a novel therapeutic target, proteinase-activated receptor (PAR)-2, in joint inflammation. Intra-articular carrageenan/kaolin (C/K) injection in mice resulted in joint swelling that was associated with synovial PAR2 up-regulation. Inhibiting receptor up-regulation using small interfering RNA technology, as confirmed by immunoblotting, substantially reduced the inflammatory response in the joint. Serine proteinase-induced joint swelling was mediated primarily via PAR2 activation, since the response to exogenous application of trypsin and tryptase was absent in PAR2 knockout mice. Furthermore, serine proteinase inhibitors were effective anti-inflammatory agents in this model. Disrupting proteolytic activation of PAR2 using antiserum (B5) directed to the receptor cleavage/activation site also attenuated C/K-induced inflammation, as did the similarly targeted PAR2 monoclonal antibody SAM-11. Finally, we report the activity of a novel small molecule PAR2 antagonist, N1-3-methylbutyryl-N4-6 aminohexanoyl-piperazine (ENMD-1068), that dose dependently attenuated joint inflammation. Our findings represent a major advance in collectively identifying PAR2 as a novel target for the future treatment of arthritis. PMID- 16260584 TI - Chronic alcohol consumption in mice increases the proportion of peripheral memory T cells by homeostatic proliferation. AB - This study examined the mechanism underlying the increase of peripheral memory phenotype T cells that occurs during chronic alcohol consumption in mice. Female C57BL/6 mice were given 20% (w/v) alcohol in the drinking water for 2 weeks to 6 months. Chronic alcohol consumption significantly induced peripheral T cell lymphopenia; up-regulated expression of CD44 on T cells and increased the percentage of CD4+CD44int/hi and CD8+CD44int/hi Ly6C+ T cells; up-regulated the expression of CD43 on CD8+ T cells; increased the percentage of interferon- producing T cells; decreased the percentage of CD8+CD28+ T cells; and down regulated the expression of CD28 on CD4+ T cells. Expression of CD25 and CD69 on peripheral CD8+ T cells was not affected and inconsistently expressed on CD4+ T cells. Neither cell type showed altered expression of CD137 or CD153. Alcohol withdrawal did not abrogate the increase in CD8+Ly6C+cells induced by alcohol consumption. In vivo bromodeoxyuridine incorporation experiments demonstrated that chronic alcohol consumption decreases naive T cells that are presumed to have emigrated from the thymus and increases proliferation of memory T cells, but accelerates peripheral T cell turnover. Together these results indicate that chronic alcohol consumption results in T cell lymphopenia, which in turn induces T cell homeostatic proliferation that increases the proportion of peripheral memory T cells relative to naive T cells. PMID- 16260583 TI - Analysis of the antinociceptive effect of the flavonoid myricitrin: evidence for a role of the L-arginine-nitric oxide and protein kinase C pathways. AB - The present study investigated the antinociceptive effects of the flavonoid myricitrin in chemical behavioral models of pain in mice and rats. Myricitrin given by i.p. or p.o. routes produced dose-related antinociception when assessed on acetic acid-induced visceral pain in mice. In addition, the i.p. administration of myricitrin exhibited significant inhibition of the neurogenic pain induced by intraplantar (i.pl.) injection of capsaicin. Like-wise, myricitrin given by i.p. route reduced the nociception produced by i.pl. injection of glutamate and phorbol myristate acetate (PMA). Western blot analysis revealed that myricitrin treatment fully prevented the protein kinase C (PKC) alpha and PKCepsilon activation by PMA in mice hind paws. Myricitrin given i.p. also inhibited the mechanical hyperalgesia induced by bradykinin, without affecting similar responses caused by epinephrine and prostaglandin E(2). The antinociception caused by myricitrin in the acetic acid test was significantly attenuated by i.p. treatment of mice with the nitric oxide precursor, L-arginine. In contrast, myricitrin antinociception was not affected by naloxone (opioid receptor antagonist) or neonatal pretreatment of mice with capsaicin and myricitrin antinociceptive effects is not related to muscle relaxant or sedative action. Together, these results indicate that myricitrin produces pronounced antinociception against chemical and mechanical models of pain in rodents. The mechanisms involved in their actions are not completely understood but seem to involve an interaction with nitric oxide-L-arginine and protein kinase C pathways. PMID- 16260585 TI - Involvement of neutrophil recruitment and protease-activated receptor 2 activation in the induction of IL-18 in mice. AB - Activated neutrophils produce serine proteases, which activate cells through protease-activated receptor 2 (PAR2). As proteinase 3 (PR3) induces the secretion of interleukin (IL)-18 from epithelial cells in combination with lipopolysaccharide (LPS) in vitro, we examined whether neutrophils, serine proteases, and PAR2 are involved in the induction of serum IL-18 and IL-18 dependent liver injury in mice treated with heat-killed Propionibacterium acnes and LPS. LPS-induced serum IL-18 levels in P. acnes-primed mice were reduced significantly by anti-Gr-1 injection (depletion of neutrophils and macrophages) but not by a macrophage "suicide" technique, using liposomes encapsulating clodronate. The IL-18 induction was decreased significantly by coadministration of a serine protease inhibitor [Nafamostat mesilate (FUT-175)] with LPS. Serum levels of tumor necrosis factor alpha and liver enzymes induced by P. acnes and LPS were abolished by anti-Gr-1 treatment, and concomitantly, liver injury (necrotic change and granuloma formation) and Gr-1(+) cell infiltration into the liver were prevented by the treatment. A deficiency of PAR2 in mice significantly impaired IL-18 induction by treatment with P. acnes and LPS, and only slight pathological changes in hepatic tissues occurred in the PAR2-deficient mice treated with P. acnes and LPS. Furthermore, coadministration of exogenous murine PR3 or a synthetic PAR2 agonist (ASKH95) with LPS in the anti-Gr-1-treated mice restored the serum IL-18 levels to those in control mice treated with P. acnes and LPS. These results indicate that neutrophil recruitment and PAR2 activation by neutrophil serine proteases are critically involved in the induction of IL-18 and IL-18-dependent liver injury in vivo. PMID- 16260586 TI - Galectin-3 interacts with naive and primed neutrophils, inducing innate immune responses. AB - The neutrophil is the first line of defense against infection. As a part of the innate immune response, neutrophils start to emigrate from blood to an affected site and their state is altered from passively circulating naive to primed, and then to fully activated. The extent of neutrophil activation and their subsequent response varies depending on the stimuli and environment that neutrophils encounter. Because neutrophils can also induce deleterious effects on host tissues, tight regulation of recruitment and functions of neutrophils is required for efficient recovery. Galectin-3, a soluble beta-galactoside binding protein, of which expression is up-regulated during inflammation/infection, is suggested to be involved in various inflammatory responses. However, the precise roles of this lectin in innate immunity remain unknown, while it has been demonstrated that galectin-3 binds to naive and primed neutrophils. Here we report that galectin-3 can induce L-selectin shedding and interleukin-8 production in naive and primed neutrophils. These activities were shown to be dependent on the presence of the C-terminal lectin domain and the N-terminal nonlectin domain of galectin-3, which is involved in oligomerization of this lectin. We also found that, after galectin-3 binds to neutrophils, primed but not naive neutrophils can cleave galectin-3, mainly through elastase, which results in the formation of truncated galectin-3 lacking the N-terminal domain. Together, these results suggest that galectin-3 activates naive and primed neutrophils, and galectin-3 activated primed neutrophils have an ability to inactivate galectin-3. PMID- 16260587 TI - Plasmacytoid dendritic cells (PDC) are the major DC subset innately producing cytokines in human lymph nodes. AB - Plasmacytoid dendritic cells (PDC) constitute a distinct subset of DC found in human peripheral lymph nodes (LN), but little is known about their function. Cell suspensions were prepared from tumor draining LN (n=20) and control LN (n=11) of women undergoing surgical resection for primary breast cancer and elective surgery for benign conditions, respectively. Using four-color flow cytometry, human leukocyte antigen-DR+ DC subsets were identified phenotypically. The proportions and numbers of cells innately producing interleukin (IL)-4, IL-10, IL 12, and interferon-gamma (IFN-gamma) were also measured from intracellular accumulation of cytokine after blocking with monensin. All flow cytometry data were collected without compensation and were compensated off-line using the Winlist algorithm (Verity software). This package also provided the subtraction program to calculate percentage positive cells and intensity of staining. PDC (CD11c-, CD123+) expressed more cytokines than did myeloid DC (CD11c+) or CD1a+ putative "migratory" DC (P<0.001). LN PDC from patients with a good prognosis (px; n=11) demonstrated a relative increase in IL-12 and IFN-gamma expression (median IL-10:IL-12 ratio=0.78 and median IL-4:IFN-gamma ratio=0.7), and PDC from LN draining poor px cancer (n=9) showed a relative increase in IL-10 and IL-4 expression (median IL-10:IL-12 ratio=1.31 and median IL-4:IFN-gamma ratio=2.6). The difference in IL-4:IFN-gamma expression between good and poor px cancer groups was significant (P<0.05). Thus, PDC innately producing cytokines were identified in cell suspensions from human LN, and the character of PDC cytokine secretion may differ between two breast cancer prognostic groups. We speculate that a shift towards PDC IL-10 and IL-4 expression could promote tumor tolerance in LN draining poor px breast cancer. PMID- 16260588 TI - Effect of fixation to the degradation of nuclear and mitochondrial DNA in different tissues. AB - Samples of different tissues were preserved in seven fixatives for periods of time extending from 1 to 336 days, to determine which fixatives reduce the time dependent degradation of DNA and preserve the histological structure. To achieve these results, three PCR systems were used: FGA and TC11 (both for nuclear DNA) and HV1 for mitochondrial DNA (mt-DNA). For long-term storage in combination with amplification of nuclear and mt-DNA, consistent results were obtained in Carnoy's solution and glutaraldehyde. Variable results were observed for buffered formalin; an mt-DNA product could be detected even after 3 months of fixation. In regard to comparison of the different tissues, the quantities recovered from skeletal muscles and kidneys were higher than from other tissues. PMID- 16260589 TI - Gene expression and localization of insulin-like growth factors and their receptors throughout amelogenesis in rat incisors. AB - Insulin-like growth factors (IGFs) are expressed in many tissues and control cell differentiation, proliferation, and apoptosis. In teeth, the temporo-spatial pattern of expression IGFs and their receptors has not been fully characterized. The purpose of this study was to obtain a comprehensive profile of their expression throughout the life cycle of ameloblasts, using the continuously erupting rat incisor model. Upper incisors of young male rats were fixed by perfusion, decalcified, and embedded in paraffin. Sections were processed for in situ hybridization and immunohistochemistry. mRNA and protein expression profiles IGF-I, IGF-II, IGF-IR, and IGF-IIR mRNA were essentially identical. At the apical loop of the incisor, very strong signals were seen in the outer enamel epithelium while the inner enamel epithelium showed a moderate reaction. In the region of ameloblasts facing pulp, inner enamel epithelium cells were still moderately reactive while signals over the outer enamel epithelium were slightly reduced. In the region of ameloblasts facing dentin and the initial portion of the secretory zone, signals in ameloblasts were weak while those over the outer enamel epithelium were strong. In the region of postsecretory transition, signals in both ameloblasts and papillary layer cells gradually increased. In maturation proper, signals in ameloblasts appeared as alternating bands of strong and weak reactivities, which corresponded to the regions of ruffle-ended and smooth-ended ameloblasts, respectively. Papillary layer cells also showed alternations in signal intensity that matched those in ameloblasts. These results suggest that the IGF family may act as an autocrine/paracrine system that influences not only cell differentiation but also the physiological activity of ameloblasts. PMID- 16260590 TI - UDP-N-acetyl-D-galactosamine: polypeptide N-acetylgalactosaminyltransferase-6 as a new immunohistochemical breast cancer marker. AB - Mucin O-glycosylation is characterized in cancer by aberrant expression of immature carbohydrate structures (Tn, T, and sialyl-Tn antigens). The UDP-N acetyl-D-galactosamine: polypeptide N-acetylgalactosaminyltransferases (ppGalNAc T) family enzymes regulate the initial steps of mucin O-glycosylation and could be responsible for the altered glycosylation observed in cancer. Considering that we recently found the ppGalNAc-T6 mRNA expressed in breast carcinomas, we produced a highly specific monoclonal antibody (MAb T6.3) to assess the expression profile of ppGalNAc-T6 protein product in breast tissues. The expression of ppGalNAc-T6 by breast carcinoma cells was confirmed on MCF-7 and T47D cell lines. In formalin-fixed tissues, ppGalNAc-T6 expression was observed in 60/74 (81%) breast cancers, 21/23 (91.3%) adjacent ductal carcinoma in situ (DCIS), 4/20 benign breast lesions (2/2 sclerosing adenosis and 2/13 fibroadenoma), and in 0/5 normal breast samples. We observed a statistically significant association of ppGalNAc-T6 expression with T1 tumor stage. This fact, as well as the observation that ppGalNAc-T6 was strongly expressed in sclerosing adenosis and in most DCIS, suggests that ppGalNAc-T6 expression could be an early event during human breast carcinogenesis. Considering that an abnormal O glycosylation greatly contributes to the phenotype and biology of breast cancer cells, ppGalNAc-T6 expression could provide new insights about breast cancer glycobiology. PMID- 16260591 TI - Zinc-secreting Paneth cells studied by ZP fluorescence. AB - We have used a new family of zinc-specific-responsive fluorescent dyes (ZPs) to study the sequestration and secretion of zinc from Paneth cells, which are located in the bases of the crypts of Lieberkuhn within the rat small intestine. Vivid ZP fluorescence zinc staining of Paneth cell secretory granules is seen in both cryostat sections and isolated crypts, providing firm evidence for a pool of labile (rapidly exchangeable) zinc within these cells. We further demonstrate that this ionic zinc pool is secreted under physiological conditions. In vivo stimulation of the small intestine by IP injection of the secretagogue pilocarpine results in discrete zinc staining within the lumens of subsequently isolated crypts, concomitant with a decrease in the zinc staining of Paneth cell granules located within the same crypts. In contrast, the secretion of zinc into the lumens of isolated crypts stimulated in vitro with either carbachol or LPS (lipopolysaccharide) is not observed. However, a distinct change in Paneth cell morphology, suggesting attempted secretion, is seen in response to the direct application of cholinergics but not LPS. These findings suggest that zinc is coreleased with other Paneth cell anti-microbials, and that the intact intestine is necessary for secretion into the crypt lumen. PMID- 16260593 TI - p38 mitogen-activated protein kinase/Hog1p regulates translation of the AU-rich element-bearing MFA2 transcript. AB - AU-rich-element (ARE)-mediated mRNA regulation occurs in Saccharomyces cerevisiae in response to external and internal stimuli through the p38 mitogen-activated protein kinase (MAPK)/Hog1p pathway. We demonstrate that the ARE-bearing MFA2 3' untranslated region (UTR) controls translation efficiency in a p38 MAPK/Hog1p dependent manner in response to carbon source growth conditions. The carbon source-regulated effect on MFA2 3'-UTR-controlled translation involves the role of conserved ARE binding proteins, the ELAV/TIA-1-like Pub1p, which can interact with the cap/eIF4G complex, and the translation/mRNA stability factor poly(A) binding protein (Pab1p). Pub1p binds the MFA2 3'-UTR in a p38 MAPK/Hog1p regulated manner in response to carbon source growth conditions. Significantly, the p38 MAPK/Hog1p is also required to modulate Pab1p in response to carbon source. We find that Pab1p can bind the MFA2 3'-UTR in a regulated manner to control MFA2 3'-UTR reporter translation. Binding of full-length Pab1p to the MFA2 3'-UTR correlates with translation repression. Importantly, Pab1p binds the MFA2 3'-UTR only in a PUB1 strain, and correlating with this requirement, Pub1p controls translation repression of MFA2 in a carbon source/Hog1p-regulated manner. These results suggest that the p38 MAPK/Hog1p pathway regulates 3'-UTR mediated translation by modulating recruitment of Pab1p and Pub1p, which can interact with the translation machinery. PMID- 16260592 TI - Interleukin-21 receptor gene induction in human T cells is mediated by T-cell receptor-induced Sp1 activity. AB - Interleukin-21 (IL-21) plays important roles in regulating the immune response. IL-21 receptor (IL-21R) mRNA is expressed at a low level in human resting T cells but is rapidly induced by mitogenic stimulation. We now investigate the basis for IL21R gene regulation in T cells. We found that the -80 to -20 region critically regulates IL-21R promoter activity and corresponds to a major DNase I hypersensitive site. Electrophoretic mobility shift assays, DNA affinity chromatography followed by mass spectrometry, and chromatin immunoprecipitation assays revealed that Sp1 binds to this region in vitro and in vivo. Moreover, mutation of the Sp1 motif markedly reduced IL-21R promoter activity, and Sp1 small interfering RNAs effectively diminished IL-21R expression in activated T cells. Interestingly, upon T-cell receptor (TCR) stimulation, T cells increased IL-21R expression and Sp1 protein levels while decreasing Sp1 phosphorylation. Moreover, phosphatase inhibitors that increased phosphorylation of Sp1 diminished IL-21R transcription. These data indicate that TCR-induced IL-21R expression is driven by TCR-mediated augmentation of Sp1 protein levels and may partly depend on the dephosphorylation of Sp1. PMID- 16260594 TI - Scavenger decapping activity facilitates 5' to 3' mRNA decay. AB - mRNA degradation occurs through distinct pathways, one primarily from the 5' end of the mRNA and the second from the 3' end. Decay from the 3' end generates the m7GpppN cap dinucleotide, which is subsequently hydrolyzed to m7Gp and ppN in Saccharomyces cerevisiae by a scavenger decapping activity termed Dcs1p. Although Dcs1p functions in the last step of mRNA turnover, we demonstrate that its activity modulates earlier steps of mRNA decay. Disruption of the DCS1 gene manifests a threefold increase of the TIF51A mRNA half-life. Interestingly, the hydrolytic activity of Dcs1p was essential for the altered mRNA turnover, as Dcs1p, but not a catalytically inactive Dcs1p mutant, complemented the increased mRNA stability. Mechanistic analysis revealed that 5' to 3' exoribonucleolytic activity was impeded in the dcs1Delta strain, resulting in the accumulation of uncapped mRNA. These data define a new role for the Dcs1p scavenger decapping enzyme and demonstrate a novel mechanism whereby the final step in the 3' mRNA decay pathway can influence 5' to 3' exoribonucleolytic activity. PMID- 16260595 TI - Repair-independent chromatin assembly onto active ribosomal genes in yeast after UV irradiation. AB - Chromatin rearrangements occur during repair of cyclobutane pyrimidine dimers (CPDs) by nucleotide excision repair (NER). Thereafter, the original structure must be restored to retain normal genomic functions. How NER proceeds through nonnucleosomal chromatin and how open chromatin is reestablished after repair are unknown. We analyzed NER in ribosomal genes (rDNA), which are present in multiple copies but only a fraction are actively transcribed and nonnucleosomal. We show that removal of CPDs is fast in the active rDNA and that chromatin reorganization occurs during NER. Furthermore, chromatin assembles on nonnucleosomal rDNA during the early events of NER but in the absence of DNA repair. The resumption of transcription after removal of CPDs correlates with the reappearance of nonnucleosomal chromatin. To date, only the passage of replication machinery was thought to package ribosomal genes in nucleosomes. In this report, we show that early events after formation of UV photoproducts in DNA also promote chromatin assembly. PMID- 16260596 TI - Xeroderma pigmentosum complementation group E protein (XPE/DDB2): purification of various complexes of XPE and analyses of their damaged DNA binding and putative DNA repair properties. AB - Xeroderma pigmentosum is characterized by increased sensitivity of the affected individuals to sunlight and light-induced skin cancers and, in some cases, to neurological abnormalities. The disease is caused by a mutation in genes XPA through XPG and the XP variant (XPV) gene. The proteins encoded by the XPA, -B, C, -D, -F, and -G genes are required for nucleotide excision repair, and the XPV gene encodes DNA polymerase eta, which carries out translesion DNA synthesis. In contrast, the mechanism by which the XPE gene product prevents sunlight-induced cancers is not known. The gene (XPE/DDB2) encodes the small subunit of a heterodimeric DNA binding protein with high affinity to UV-damaged DNA (UV damaged DNA binding protein [UV-DDB]). The DDB2 protein exists in at least four forms in the cell: monomeric DDB2, DDB1-DDB2 heterodimer (UV-DDB), and as a protein associated with both the Cullin 4A (CUL4A) complex and the COP9 signalosome. To better define the role of DDB2 in the cellular response to DNA damage, we purified all four forms of DDB2 and analyzed their DNA binding properties and their effects on mammalian nucleotide excision repair. We find that DDB2 has an intrinsic damaged DNA binding activity and that under our assay conditions neither DDB2 nor complexes that contain DDB2 (UV-DDB, CUL4A, and COP9) participate in nucleotide excision repair carried out by the six-factor human excision nuclease. PMID- 16260597 TI - Mixed-disulfide folding intermediates between thyroglobulin and endoplasmic reticulum resident oxidoreductases ERp57 and protein disulfide isomerase. AB - We present the first identification of transient folding intermediates of endogenous thyroglobulin (Tg; a large homodimeric secretory glycoprotein of thyrocytes), which include mixed disulfides with endogenous oxidoreductases servicing Tg folding needs. Formation of disulfide-linked Tg adducts with endoplasmic reticulum (ER) oxidoreductases begins cotranslationally. Inhibition of ER glucosidase activity blocked formation of a subgroup of Tg adducts containing ERp57 while causing increased Tg adduct formation with protein disulfide isomerase (PDI), delayed adduct resolution, perturbed oxidative folding of Tg monomers, impaired Tg dimerization, increased Tg association with BiP/GRP78 and GRP94, activation of the unfolded protein response, increased ER-associated degradation of a subpopulation of Tg, partial Tg escape from ER quality control with increased secretion of free monomers, and decreased overall Tg secretion. These data point towards mixed disulfides with the ERp57 oxidoreductase in conjunction with calreticulin/calnexin chaperones acting as normal early Tg folding intermediates that can be "substituted" by PDI adducts only at the expense of lower folding efficiency with resultant ER stress. PMID- 16260598 TI - TRAF6 is required for TRAF2-dependent CD40 signal transduction in nonhemopoietic cells. AB - The emerging role of CD40, a tumor necrosis factor (TNF) receptor family member, in immune regulation, disease pathogenesis, and cancer therapy necessitates the analysis of CD40 signal transduction in a wide range of tissue types. In this study we present evidence that the CD40-interacting proteins TRAF2 and TRAF6 play an important physiological role in CD40 signaling in nonhemopoietic cells. Using mutational analysis of the CD40 cytoplasmic tail, we demonstrate that the specific binding of TRAF2 to CD40 is required for efficient signaling on the NF kappaB, Jun N-terminal protein kinase (JNK), and p38 axis. In fibroblasts lacking TRAF2 or in carcinoma cells in which TRAF2 has been depleted by RNA interference, the CD40-mediated activation of NF-kappaB and JNK is significantly reduced, and the activation of p38 and Akt is severely impaired. Interestingly, whereas the TRAF6-interacting membrane-proximal domain of CD40 has a minor role in signal transduction, studies utilizing TRAF6 knockout fibroblasts and RNA interference in epithelial cells reveal that the CD40-induced activation of NF-kappaB, JNK, p38, and Akt requires the integrity of TRAF6. Furthermore, we provide evidence that TRAF6 regulates CD40 signal transduction not only through its direct binding to CD40 but also indirectly via its association with TRAF2. These observations provide novel insight into the mechanisms of CD40 signaling and the multiple roles played by TRAF6 in signal transduction. PMID- 16260599 TI - A novel mitogen-activated protein kinase docking site in the N terminus of MEK5alpha organizes the components of the extracellular signal-regulated kinase 5 signaling pathway. AB - The alternative splicing of the mek5 gene gives rise to two isoforms. MEK5beta lacks an extended N terminus present in MEK5alpha. Comparison of their activities led us to identify a novel mitogen-activated protein kinase (MAPK) docking site in the N terminus of MEK5alpha that is distinct from the consensus motif identified in the other MAPK kinases. It consists of a cluster of acidic residues at position 61 and positions 63 to 66. The formation of the MEK5/extracellular signal-regulated kinase 5 (ERK5) complex is critical for MEK5 to activate ERK5, to increase transcription via MEF2, and to enhance cellular survival in response to osmotic stress. Certain mutations in the ERK5 docking site that prevent MEK5/ERK5 interaction also abrogate the ability of MEKK2 to bind and activate MEK5. However, the identification of MEK5alpha mutants with selective binding defect demonstrates that the MEK5/ERK5 interaction does not rely on the binding of MEK5alpha to MEKK2 via their respective PB1 domains. Altogether these results establish that the N terminus of MEK5alpha is critical for the specific organization of the components of the ERK5 signaling pathway. PMID- 16260600 TI - Convergence of protein kinase C and JAK-STAT signaling on transcription factor GATA-4. AB - Angiotensin II (AII), a potent vasoactive hormone, acts on numerous organs via G protein-coupled receptors and elicits cell-specific responses. At the level of the heart, AII stimulation alters gene transcription and leads to cardiomyocyte hypertrophy. Numerous intracellular signaling pathways are activated in this process; however, which of these directly link receptor activation to transcriptional regulation remains undefined. We used the atrial natriuretic factor (ANF) gene (NPPA) as a marker to elucidate the signaling cascades involved in AII transcriptional responses. We show that ANF transcription is activated directly by the AII type 1 receptor and precedes the development of myocyte hypertrophy. This response maps to STAT and GATA binding sites, and the two elements transcriptionally cooperate to mediate signaling through the JAK-STAT and protein kinase C (PKC)-GATA-4 pathways. PKC phosphorylation enhances GATA-4 DNA binding activity, and STAT-1 functionally and physically interacts with GATA 4 to synergistically activate AII and other growth factor-inducible promoters. Moreover, GATA factors are able to recruit STAT proteins to target promoters via GATA binding sites, which are sufficient to support synergy. Thus, STAT proteins can act as growth factor-inducible coactivators of tissue-specific transcription factors. Interactions between STAT and GATA proteins may provide a general paradigm for understanding cell specificity of cytokine and growth factor signaling. PMID- 16260601 TI - Kinetic analysis of Smad nucleocytoplasmic shuttling reveals a mechanism for transforming growth factor beta-dependent nuclear accumulation of Smads. AB - Upon transforming growth factor beta (TGF-beta) stimulation, Smads accumulate in the nucleus, where they regulate gene expression. Using fluorescence perturbation experiments on Smad2 and Smad4 fused to either enhanced green fluorescent protein or photoactivatable green fluorescent protein, we have studied the kinetics of Smad nucleocytoplasmic shuttling in a quantitative manner in vivo. We have obtained rate constants for import and export of Smad2 and show that the cytoplasmic localization of Smad2 in uninduced cells reflects its nuclear export being more rapid than import. We find that TGF-beta-induced nuclear accumulation of Smad2 is caused by a pronounced drop in the export rate of Smad2 from the nucleus, which is associated with a strong decrease in nuclear mobility of Smad2 and Smad4. TGF-beta-induced nuclear accumulation involves neither a release from cytoplasmic retention nor an increase in Smad2 import rate. Hence, TGF-beta dependent nuclear accumulation of Smad2 is caused exclusively by selective nuclear trapping of phosphorylated, complexed Smad2. The proposed mechanism reconciles signal-dependent nuclear accumulation of Smad2 with its continuous nucleocytoplasmic cycling properties. PMID- 16260602 TI - The novel ATP-binding cassette protein ARB1 is a shuttling factor that stimulates 40S and 60S ribosome biogenesis. AB - ARB1 is an essential yeast protein closely related to members of a subclass of the ATP-binding cassette (ABC) superfamily of proteins that are known to interact with ribosomes and function in protein synthesis or ribosome biogenesis. We show that depletion of ARB1 from Saccharomyces cerevisiae cells leads to a deficit in 18S rRNA and 40S subunits that can be attributed to slower cleavage at the A0, A1, and A2 processing sites in 35S pre-rRNA, delayed processing of 20S rRNA to mature 18S rRNA, and a possible defect in nuclear export of pre-40S subunits. Depletion of ARB1 also delays rRNA processing events in the 60S biogenesis pathway. We further demonstrate that ARB1 shuttles from nucleus to cytoplasm, cosediments with 40S, 60S, and 80S/90S ribosomal species, and is physically associated in vivo with TIF6, LSG1, and other proteins implicated previously in different aspects of 60S or 40S biogenesis. Mutations of conserved ARB1 residues expected to function in ATP hydrolysis were lethal. We propose that ARB1 functions as a mechanochemical ATPase to stimulate multiple steps in the 40S and 60S ribosomal biogenesis pathways. PMID- 16260603 TI - Smooth muscle-specific genes are differentially sensitive to inhibition by Elk-1. AB - Understanding the mechanism of smooth muscle cell (SMC) differentiation will provide the foundation for elucidating SMC-related diseases, such as atherosclerosis, restenosis, and asthma. In the current study, overexpression of Elk-1 in SMCs down-regulated expression of several endogenous smooth muscle restricted proteins, including telokin, SM22alpha, and smooth muscle alpha-actin. In contrast, down-regulation of endogenous Elk-1 in smooth muscle cells increased the expression of only telokin and SM22alpha, suggesting that smooth muscle specific promoters are differentially sensitive to the inhibitory effects of Elk 1. Consistent with this, overexpression of the DNA binding domain of Elk-1, which acts as a dominant-negative protein by displacing endogenous Elk-1, enhanced the expression of telokin and SM22alpha without affecting expression of smooth muscle alpha-actin. Elk-1 suppressed the activity of smooth muscle-restricted promoters, including the telokin promoter that does not contain a consensus Elk-1 binding site, through its ability to block myocardin-induced activation of the promoters. Gel mobility shift and chromatin immunoprecipitation assays revealed that Elk-1 binds to a nonconsensus binding site in the telokin promoter and Elk-1 binding is dependent on serum response factor (SRF) binding to a nearby CArG box. Although overexpression of the SRF-binding B-box domain of Elk-1 is sufficient to repress the myocardin activation of the telokin promoter, this repression is not as complete as that seen with an Elk-1 fragment that includes the DNA binding domain. In addition, reporter gene assays demonstrate that an intact Elk-1 binding site in the telokin promoter is required for Elk-1 to maximally inhibit promoter activity. Together, these data suggest that the differential sensitivity of smooth muscle-specific genes to inhibition by Elk-1 may play a role in the complex changes in smooth muscle-specific protein expression that are observed under pathological conditions. PMID- 16260604 TI - The histone fold subunits of Drosophila CHRAC facilitate nucleosome sliding through dynamic DNA interactions. AB - The chromatin accessibility complex (CHRAC) is an abundant, evolutionarily conserved nucleosome remodeling machinery able to catalyze histone octamer sliding on DNA. CHRAC differs from the related ACF complex by the presence of two subunits with molecular masses of 14 and 16 kDa, whose structure and function were not known. We determined the structure of Drosophila melanogaster CHRAC14 CHRAC16 by X-ray crystallography at 2.4-angstroms resolution and found that they dimerize via a variant histone fold in a typical handshake structure. In further analogy to histones, CHRAC14-16 contain unstructured N- and C-terminal tail domains that protrude from the handshake structure. A dimer of CHRAC14-16 can associate with the N terminus of ACF1, thereby completing CHRAC. Low-affinity interactions of CHRAC14-16 with DNA significantly improve the efficiency of nucleosome mobilization by limiting amounts of ACF. Deletion of the negatively charged C terminus of CHRAC16 enhances DNA binding 25-fold but leads to inhibition of nucleosome sliding, in striking analogy to the effect of the DNA chaperone HMGB1 on nucleosome sliding. The presence of a surface compatible with DNA interaction and the geometry of an H2A-H2B heterodimer may provide a transient acceptor site for DNA dislocated from the histone surface and therefore facilitate the nucleosome remodeling process. PMID- 16260605 TI - Human c-Myc isoforms differentially regulate cell growth and apoptosis in Drosophila melanogaster. AB - The human c-myc proto-oncogene, implicated in the control of many cellular processes including cell growth and apoptosis, encodes three isoforms which differ in their N-terminal region. The functions of these isoforms have never been addressed in vivo. Here, we used Drosophila melanogaster to examine their functions in a fully integrated system. First, we established that the human c Myc protein can rescue lethal mutations of the Drosophila myc ortholog, dmyc, demonstrating the biological relevance of this model. Then, we characterized a new lethal dmyc insertion allele, which permits expression of human c-Myc in place of dMyc and used it to compare physiological activities of these isoforms in whole-organism rescue, transcription, cell growth, and apoptosis. These isoforms differ both quantitatively and qualitatively. Most remarkably, while the small c-MycS form truncated for much of its N-terminal trans-activation domain efficiently rescued viability and cell growth, it did not induce detectable programmed cell death. Our data indicate that the main functional difference between c-Myc isoforms resides in their apoptotic properties and that the N terminal region, containing the conserved MbI motif, is decisive in governing the choice between growth and death. PMID- 16260606 TI - Protein phosphatase 5 is required for ATR-mediated checkpoint activation. AB - In response to DNA damage or replication stress, the protein kinase ATR is activated and subsequently transduces genotoxic signals to cell cycle control and DNA repair machinery through phosphorylation of a number of downstream substrates. Very little is known about the molecular mechanism by which ATR is activated in response to genotoxic insults. In this report, we demonstrate that protein phosphatase 5 (PP5) is required for the ATR-mediated checkpoint activation. PP5 forms a complex with ATR in a genotoxic stress-inducible manner. Interference with the expression or the activity of PP5 leads to impairment of the ATR-mediated phosphorylation of hRad17 and Chk1 after UV or hydroxyurea treatment. Similar results are obtained in ATM-deficient cells, suggesting that the observed defect in checkpoint signaling is the consequence of impaired functional interaction between ATR and PP5. In cells exposed to UV irradiation, PP5 is required to elicit an appropriate S-phase checkpoint response. In addition, loss of PP5 leads to premature mitosis after hydroxyurea treatment. Interestingly, reduced PP5 activity exerts differential effects on the formation of intranuclear foci by ATR and replication protein A, implicating a functional role for PP5 in a specific stage of the checkpoint signaling pathway. Taken together, our results suggest that PP5 plays a critical role in the ATR-mediated checkpoint activation. PMID- 16260607 TI - CRMP-2 is involved in kinesin-1-dependent transport of the Sra-1/WAVE1 complex and axon formation. AB - A neuron has two types of highly polarized cell processes, the single axon and multiple dendrites. One of the fundamental questions of neurobiology is how neurons acquire such specific and polarized morphologies. During neuronal development, various actin-binding proteins regulate dynamics of actin cytoskeleton in the growth cones of developing axons. The regulation of actin cytoskeleton in the growth cones is thought to be involved in axon outgrowth and axon-dendrite specification. However, it is largely unknown which actin-binding proteins are involved in axon-dendrite specification and how they are transported into the developing axons. We have previously reported that collapsin response mediator protein 2 (CRMP-2) plays a critical role in axon outgrowth and axon dendrite specification (N. Inagaki, K. Chihara, N. Arimura, C. Menager, Y. Kawano, N. Matsuo, T. Nishimura, M. Amano, and K. Kaibuchi, Nat. Neurosci. 4:781 782, 2001). Here, we found that CRMP-2 interacted with the specifically Rac1 associated protein 1 (Sra-1)/WASP family verprolin-homologous protein 1 (WAVE1) complex, which is a regulator of actin cytoskeleton. The knockdown of Sra-1 and WAVE1 by RNA interference canceled CRMP-2-induced axon outgrowth and multiple axon formation in cultured hippocampal neurons. We also found that CRMP-2 interacted with the light chain of kinesin-1 and linked kinesin-1 to the Sra 1/WAVE1 complex. The knockdown of CRMP-2 and kinesin-1 delocalized Sra-1 and WAVE1 from the growth cones of axons. These results suggest that CRMP-2 transports the Sra-1/WAVE1 complex to axons in a kinesin-1-dependent manner and thereby regulates axon outgrowth and formation. PMID- 16260609 TI - Regulation of apoptotic c-Jun N-terminal kinase signaling by a stabilization based feed-forward loop. AB - A sequential kinase cascade culminating in activation of c-Jun N-terminal kinases (JNKs) plays a fundamental role in promoting apoptotic death in many cellular contexts. The mechanisms by which this pathway is engaged in response to apoptotic stimuli and suppressed in viable cells are largely unknown. Here, we show that apoptotic stimuli increase endogenous cellular levels of pathway components, including POSH, mixed lineage kinases (MLKs), and JNK interacting protein 1, and that this effect occurs through protein stabilization and requires the presence of POSH as well as activation of MLKs and JNKs. Our findings suggest a self-amplifying, feed-forward loop mechanism by which apoptotic stimuli promote the stabilization of JNK pathway components, thereby contributing to cell death. PMID- 16260608 TI - The DnaJ-related factor Mrj interacts with nuclear factor of activated T cells c3 and mediates transcriptional repression through class II histone deacetylase recruitment. AB - The calcium-regulated protein phosphatase calcineurin (PP2B) functions as a regulator of gene expression in diverse tissues through the dephosphorylation and activation of a family of transcription factors known as nuclear factor of activated T cells (NFAT). Here we show that NFATc3, in addition to being calcium responsive, is regulated through an indirect recruitment of class II histone deacetylases (HDACs). Specifically, yeast two-hybrid screening with the rel homology domain of NFATc3 identified the chaperone mammalian relative of DnaJ (Mrj) as a specific interacting factor. Mrj and NFATc3 were shown to directly associate with one another in mammalian cells and in vitro. Mrj served as a potent inhibitor of NFAT transcriptional activity within the nucleus through a mechanism involving histone deacetylase recruitment in conjunction with heat shock stimulation. Indeed, Mrj was determined to interact with class II histone deacetylases, each of which translocated to the nucleus following heat shock stimulation. Mrj also decreased NFATc3 occupancy of the tumor necrosis factor alpha promoter in cardiomyocytes in an HDAC-dependent manner, and Mrj blocked calcineurin-induced cardiomyocyte hypertrophic growth. Conversely, small interfering-RNA-mediated reduction of Mrj augmented NFAT transcriptional activity and spontaneously induced cardiac myocyte growth. Collectively, our results define a novel response pathway whereby NFATc3 is negatively regulated by class II histone deacetylases through the DnaJ (heat shock protein-40) superfamily member Mrj. PMID- 16260610 TI - Menin is a regulator of the stress response in Drosophila melanogaster. AB - Menin, the product of the multiple endocrine neoplasia type I gene, has been implicated in several biological processes, including the control of gene expression and apoptosis, the modulation of mitogen-activated protein kinase pathways, and DNA damage sensing or repair. In this study, we have investigated the function of menin in the model organism Drosophila melanogaster. We show that Drosophila lines overexpressing menin or an RNA interference for this gene develop normally but are impaired in their response to several stresses, including heat shock, hypoxia, hyperosmolarity and oxidative stress. In the embryo subjected to heat shock, this impairment was characterized by a high degree of developmental arrest and lethality. The overexpression of menin enhanced the expression of HSP70 in embryos and interfered with its down regulation during recovery at the normal temperature. In contrast, the inhibition of menin with RNA interference reduced the induction of HSP70 and blocked the activation of HSP23 upon heat shock, Menin was recruited to the Hsp70 promoter upon heat shock and menin overexpression stimulated the activity of this promoter in embryos. A 70-kDa inducible form of menin was expressed in response to heat shock, indicating that menin is also regulated in conditions of stress. The induction of HSP70 and HSP23 was markedly reduced or absent in mutant embryos harboring a deletion of the menin gene. These embryos, which did not express the heat shock-inducible form of menin, were also hypersensitive to various conditions of stress. These results suggest a novel role for menin in the control of the stress response and in processes associated with the maintenance of protein integrity. PMID- 16260612 TI - Cyclin D3 promotes adipogenesis through activation of peroxisome proliferator activated receptor gamma. AB - In addition to their role in cell cycle progression, new data reveal an emerging role of D-type cyclins in transcriptional regulation and cellular differentiation processes. Using 3T3-L1 cell lines to study adipogenesis, we observed an up regulation of cyclin D3 expression throughout the differentiation process. Surprisingly, cyclin D3 was only minimally expressed during the initial stages of adipogenesis, when mitotic division is prevalent. This seemingly paradoxical expression led us to investigate a potential cell cycle-independent role for cyclin D3 during adipogenesis. We show here a direct interaction between cyclin D3 and the nuclear receptor peroxisome proliferator-activated receptor gamma (PPARgamma). Our experiments reveal cyclin D3 acts as a ligand-dependent PPARgamma coactivator, which, together with its cyclin-dependent kinase partner, phosphorylates the A-B domain of the nuclear receptor. Overexpression and knockdown studies with cyclin D3 had marked effects on PPARgamma activity and subsequently on adipogenesis. Chromatin immunoprecipitation assays confirm the participation of cyclin D3 in the regulation of PPARgamma target genes. We show that cyclin D3 mutant mice are protected from diet-induced obesity, display smaller adipocytes, have reduced adipogenic gene expression, and are insulin sensitive. Our results indicate that cyclin D3 is an important factor governing adipogenesis and obesity. PMID- 16260611 TI - Phosphorylation by Rho kinase regulates CRMP-2 activity in growth cones. AB - Collapsin response mediator protein 2 (CRMP-2) enhances the advance of growth cones by regulating microtubule assembly and Numb-mediated endocytosis. We previously showed that Rho kinase phosphorylates CRMP-2 during growth cone collapse; however, the roles of phosphorylated CRMP-2 in growth cone collapse remain to be clarified. Here, we report that CRMP-2 phosphorylation by Rho kinase cancels the binding activity to the tubulin dimer, microtubules, or Numb. CRMP-2 binds to actin, but its binding is not affected by phosphorylation. Electron microscopy revealed that CRMP-2 localizes on microtubules, clathrin-coated pits, and actin filaments in dorsal root ganglion neuron growth cones, while phosphorylated CRMP-2 localizes only on actin filaments. The phosphomimic mutant of CRMP-2 has a weakened ability to enhance neurite elongation. Furthermore, ephrin-A5 induces phosphorylation of CRMP-2 via Rho kinase during growth cone collapse. Taken together, these results suggest that Rho kinase phosphorylates CRMP-2, and inactivates the ability of CRMP-2 to promote microtubule assembly and Numb-mediated endocytosis, during growth cone collapse. PMID- 16260613 TI - A nuclear surveillance pathway for mRNAs with defective polyadenylation. AB - The pap1-5 mutation in poly(A) polymerase causes rapid depletion of mRNAs at restrictive temperatures. Residual mRNAs are polyadenylated, indicating that Pap1 5p retains at least partial activity. In pap1-5 strains lacking Rrp6p, a nucleus specific component of the exosome complex of 3'-5' exonucleases, accumulation of poly(A)+ mRNA was largely restored and growth was improved. The catalytically inactive mutant Rrp6-1p did not increase growth of the pap1-5 strain and conferred much less mRNA stabilization than rrp6delta. This may indicate that the major function of Rrp6p is in RNA surveillance. Inactivation of core exosome components, Rrp41p and Mtr3p, or the nuclear RNA helicase Mtr4p gave different phenotypes, with accumulation of deadenylated and 3'-truncated mRNAs. We speculate that slowed mRNA polyadenylation in the pap1-5 strain is detected by a surveillance activity of Rrp6p, triggering rapid deadenylation and exosome mediated degradation. In wild-type strains, assembly of the cleavage and polyadenylation complex might be suboptimal at cryptic polyadenylation sites, causing slowed polyadenylation. PMID- 16260614 TI - Homologues of the Caenorhabditis elegans Fox-1 protein are neuronal splicing regulators in mammals. AB - A vertebrate homologue of the Fox-1 protein from C. elegans was recently shown to bind to the element GCAUG and to act as an inhibitor of alternative splicing patterns in muscle. The element UGCAUG is a splicing enhancer element found downstream of numerous neuron-specific exons. We show here that mouse Fox-1 (mFox 1) and another homologue, Fox-2, are both specifically expressed in neurons in addition to muscle and heart. The mammalian Fox genes are very complex transcription units that generate transcripts from multiple promoters and with multiple internal exons whose inclusion is regulated. These genes produce a large family of proteins with variable N and C termini and internal deletions. We show that the overexpression of both Fox-1 and Fox-2 isoforms specifically activates splicing of neuronally regulated exons. This splicing activation requires UGCAUG enhancer elements. Conversely, RNA interference-mediated knockdown of Fox protein expression inhibits splicing of UGCAUG-dependent exons. These experiments show that this large family of proteins regulates splicing in the nervous system. They do this through a splicing enhancer function, in addition to their apparent negative effects on splicing in vertebrate muscle and in worms. PMID- 16260615 TI - Transforming growth factor beta-dependent sequential activation of Smad, Bim, and caspase-9 mediates physiological apoptosis in gastric epithelial cells. AB - Transforming growth factor beta (TGF-beta) has been implicated in the maintenance of homeostasis in various organs, including the gastric epithelium. In particular, TGF-beta-induced signaling was shown to be required for the differentiation-associated physiological apoptosis of gastric epithelial cells, but its mechanism has not been well understood. In this study, the molecular mechanism of TGF-beta-induced apoptosis was analyzed in a human gastric epithelial cell line, SNU16, as an in vitro model. Expression of Smad7 and Bcl X(L), but not viral FLIP, was shown to prevent TGF-beta-induced apoptosis, indicating an exclusive requirement of the activation of Smad signaling pathway and mitochondrial dysfunction followed by activation of caspase-9. In addition, treatment with TGF-beta induced binding of Bim, a proapoptotic Bcl-2 homology domain 3 (BH3)-only protein, to Bcl-X(L), which is dependent on the activation of Smad, and reduction in the expression of Bim by RNA interference decreased the sensitivity to TGF-beta-induced apoptosis. Moreover, we found abnormalities in the gastric epithelium of both Bim and caspase-9 knockout mice; these abnormalities were associated with a defect of physiological apoptosis in gastric epithelial cells. These results indicate for the first time that TGF-beta is involved in the physiological loss of gastric epithelial cells by activating apoptosis mediated by Smad, Bim, and caspase-9. PMID- 16260616 TI - The basic domain of ATH5 mediates neuron-specific promoter activity during retina development. AB - In the developing retina, the gene encoding the beta3 subunit of the neuronal nicotinic receptor, a specific marker of retinal ganglion cells, is under the direct control of the atonal homolog 5 (ATH5) basic helix-loop-helix (bHLH) transcription factor. Although quite short (143 bp in length), the beta3 promoter has the remarkable capacity to discriminate between ATH5 and the other neuronal bHLH proteins expressed in the developing nervous system. We have identified three amino acids within the basic domain that confer specificity to the ATH5 protein. These residues do not mediate direct DNA binding but are required for interaction between ATH5 and chromatin-associated proteins during retina development. When misexpressed in neurons, the myogenic bHLH factor MyoD is also able to activate the beta3 gene. This, however, is achieved not by binding of the protein to the promoter but by dimerization of MyoD with a partner, a process that depends not on the basic domain but on the HLH domain. By sequestering an E box-binding protein, MyoD relieves the active repression that blocks the beta3 promoter in most neurons. The mechanisms used by bHLH proteins to activate beta3 thus highlight how ATH5 is selected by the beta3 promoter and coordinates the derepression and transcriptional activation of the beta3 gene during the specification of retinal ganglion cells. PMID- 16260617 TI - Intrinsic function of the aryl hydrocarbon (dioxin) receptor as a key factor in female reproduction. AB - Dioxins exert a variety of adverse effects on organisms, including teratogenesis, immunosuppression, tumor promotion, and estrogenic action. Studies using aryl hydrocarbon receptor (AhR)-deficient mice suggest that the majority of these toxic effects are mediated by the AhR. In spite of the adverse effects mediated by this receptor, the AhR gene is conserved among a number of animal species, ranging from invertebrates to vertebrates. This high degree of conservation strongly suggests that AhR possesses an important physiologic function, and a critical function is also supported by the reduced fertility observed with AhR null female mice. We demonstrate that AhR plays a crucial role in female reproduction by regulating the expression of ovarian P450 aromatase (Cyp19), a key enzyme in estrogen synthesis. As revealed by in vitro reporter gene assay and in vivo chromatin immunoprecipitation assay, AhR cooperates with an orphan nuclear receptor, Ad4BP/SF-1, to activate Cyp19 gene transcription in ovarian granulosa cells. Administration to female mice of an AhR ligand, DMBA (9,10 dimethyl-1,2-benzanthracene), induced ovarian Cyp19 gene expression, irrespective of the intrinsic phase of the estrus cycle. In addition to elucidating a physiological function for AhR, our studies also suggest a possible mechanism for the toxic effects of exogenous AhR ligands as endocrine disruptors. PMID- 16260618 TI - Loss of the putative tumor suppressor band 4.1B/Dal1 gene is dispensable for normal development and does not predispose to cancer. AB - The band 4.1 proteins are cytoskeletal proteins, harboring a conserved FERM domain highly homologous to the N-terminal FERM domain of ezrin, radixin, moesin, and merlin. Recently, a truncated form of the 4.1B protein, termed Dal-1, was identified in a screen as down regulated in adenocarcinoma of the lung and was mapped to chromosome 18p11.3, which is lost in 38% of primary non-small cell lung carcinoma tumors. Analysis of several meningiomas has shown that Dal-1 expression was lost in 76% of the tumors. To further elucidate the function of the 4.1B/Dal 1 gene in development and tumorigenesis we generated mice deficient for this allele. The 4.1B/Dal-1 null mice develop normally and are fertile. Rates of cellular proliferation and apoptosis in brain, mammary, and lung tissues from the 4.1B/Dal-1 null mice were indistinguishable from those seen with wild-type mice. Aging studies indicate that these mice do not have a propensity to develop tumors. Analysis of fibroblasts from these mice demonstrated that the growth characteristics and kinetics of these cells were not different from those of cells from the wild-type mice. These findings indicate that the 4.1B gene is not required for normal development and that 4.1B/Dal-1 does not function as a tumor suppressor gene. PMID- 16260619 TI - Insights into the role of histone H3 and histone H4 core modifiable residues in Saccharomyces cerevisiae. AB - The biological significance of recently described modifiable residues in the globular core of the bovine nucleosome remains elusive. We have mapped these modification sites onto the Saccharomyces cerevisiae histones and used a genetic approach to probe their potential roles both in heterochromatic regions of the genome and in the DNA repair response. By mutating these residues to mimic their modified and unmodified states, we have generated a total of 39 alleles affecting 14 residues in histones H3 and H4. Remarkably, despite the apparent evolutionary pressure to conserve these near-invariant histone amino acid sequences, the vast majority of mutant alleles are viable. However, a subset of these variant proteins elicit an effect on transcriptional silencing both at the ribosomal DNA locus and at telomeres, suggesting that posttranslational modification(s) at these sites regulates formation and/or maintenance of heterochromatin. Furthermore, we provide direct mass spectrometry evidence for the existence of histone H3 K56 acetylation in yeast. We also show that substitutions at histone H4 K91, K59, S47, and R92 and histone H3 K56 and K115 lead to hypersensitivity to DNA-damaging agents, linking the significance of the chemical identity of these modifiable residues to DNA metabolism. Finally, we allude to the possible molecular mechanisms underlying the effects of these modifications. PMID- 16260620 TI - Snm1-deficient mice exhibit accelerated tumorigenesis and susceptibility to infection. AB - The eukaryotic SNM1 gene family has been implicated in a number of cellular pathways, including repair of DNA interstrand cross-links, involvement in VDJ recombination, repair of DNA double-strand breaks, and participation in cell cycle checkpoint pathways. In particular, mammalian SNM1 has been shown to be required in a mitotic checkpoint that causes arrest of cells in prophase prior to chromosome condensation in response to spindle poisons. Here, we report on the phenotype of a knockout of Snm1 in the mouse. Snm1-/- mice are viable and fertile but exhibit a complex phenotype. Both homozygous and heterozygous mice show a decline in survival compared to wild-type littermates. In homozygous mutant males, this reduction in survival is principally due to bacterial infections in the preputial and mandibular glands and to a lesser extent to tumorigenesis, while in homozygous and heterozygous females, it is due almost solely to tumorigenesis. The high incidence of bacterial infections in the homozygous mutant males suggests an immune dysfunction; however, examinations of T- and B cell development and immunoglobulin class switching did not reveal a defect in these pathways. Crossing of Snm1 mutant mice with a Trp53 null mutant resulted in an increase in mortality and a restriction of the tumor type to lymphomas, particularly those of the thymus. Taken together, these findings demonstrate that Snm1 is a tumor suppressor in mice that in addition has a role in immunity. PMID- 16260621 TI - 53BP1 cooperates with p53 and functions as a haploinsufficient tumor suppressor in mice. AB - p53 binding protein 1 (53BP1) is a putative DNA damage sensor that accumulates at sites of double-strand breaks (DSBs) in a manner dependent on histone H2AX. Here we show that the loss of one or both copies of 53BP1 greatly accelerates lymphomagenesis in a p53-null background, suggesting that 53BP1 and p53 cooperate in tumor suppression. A subset of 53BP1-/- p53-/- lymphomas, like those in H2AX-/ p53-/- mice, were diploid and harbored clonal translocations involving antigen receptor loci, indicating misrepair of DSBs during V(D)J recombination as one cause of oncogenic transformation. Loss of a single 53BP1 allele compromised genomic stability and DSB repair, which could explain the susceptibility of 53BP1+/- mice to tumorigenesis. In addition to structural aberrations, there were high rates of chromosomal missegregation and accumulation of aneuploid cells in 53BP1-/- p53+/+ and 53BP1-/- p53-/- tumors as well as in primary 53BP1-/- splenocytes. We conclude that 53BP1 functions as a dosage-dependent caretaker that promotes genomic stability by a mechanism that preserves chromosome structure and number. PMID- 16260622 TI - Caveolin-1 is not essential for biosynthetic apical membrane transport. AB - Caveolin-1 has been implicated in apical transport of glycosylphosphatidylinositol (GPI)-anchored proteins and influenza virus hemagglutinin (HA). Here we have studied the role of caveolin-1 in apical membrane transport by generating caveolin-1-deficient Madin-Darby canine kidney (MDCK) cells using retrovirus-mediated RNA interference. The caveolin-1 knockdown (cav1-KD) MDCK cells were devoid of caveolae. In addition, caveolin-2 was retained in the Golgi apparatus in cav1-KD MDCK cells. However, we found no significant alterations in the apical transport kinetics of GPI-anchored proteins or HA upon depletion of caveolin-1. Similar results were obtained using embryonic fibroblasts from caveolin-1-knockout mice. Thus, we conclude that caveolin-1 does not play a major role in lipid raft-mediated biosynthetic membrane trafficking. PMID- 16260624 TI - Exon selection in alpha-tropomyosin mRNA is regulated by the antagonistic action of RBM4 and PTB. AB - RNA-binding motif protein 4 (RBM4) has been implicated in the regulation of precursor mRNA splicing. Using differential display analysis, we identified mRNAs that associate with RBM4-containing messenger RNPs in vivo. Among these mRNAs, alpha-tropomyosin (alpha-TM) is known to exhibit a muscle cell type-specific splicing pattern. The level of the skeletal muscle-specific alpha-TM mRNA isoform partially correlated with that of RBM4 in human tissues examined and could be modulated by ectopic overexpression or suppression of RBM4. These results indicated that RBM4 directly influences the expression of the skeletal muscle specific alpha-TM isoform. Using minigenes, we demonstrated that RBM4 can activate the selection of skeletal muscle-specific exons, possibly via binding to intronic pyrimidine-rich elements. By contrast, the splicing regulator polypyrimidine tract binding protein (PTB) excluded these exons; moreover, RBM4 antagonized this PTB-mediated exon exclusion likely by competing with PTB for binding to a CU-rich element. This study suggests a possible mechanism underlying the regulated alternative splicing of alpha-TM by the antagonistic splicing regulators RBM4 and PTB. PMID- 16260623 TI - Tumor-derived p53 mutants induce NF-kappaB2 gene expression. AB - Overexpression of mutant p53 is a common theme in tumors, suggesting a selective pressure for p53 mutation in cancer development and progression. To determine how mutant p53 expression may lead to survival advantage in human cancer cells, we generated stable cell lines expressing p53 mutants p53-R175H, -R273H, and -D281G by use of p53-null human H1299 (lung carcinoma) cells. Compared to vector transfected cells, H1299 cells expressing mutant p53 showed a survival advantage when treated with etoposide, a common chemotherapeutic agent; however, cells expressing the transactivation-deficient triple mutant p53-D281G (L22Q/W23S) had significantly lower resistance to etoposide. Gene expression profiling of cells expressing transcriptionally active mutant p53 proteins revealed the striking pattern that all three p53 mutants induced expression of approximately 100 genes involved in cell growth, survival, and adhesion. The gene NF-kappaB2 is a prominent member of this group, whose overexpression in H1299 cells also leads to chemoresistance. Treatment of H1299 cells expressing p53-R175H with small interfering RNA specific for NF-kappaB2 made these cells more sensitive to etoposide. We have also observed activation of the NF-kappaB2 pathway in mutant p53-expressing cells. Thus, one possible pathway through which mutants of p53 may induce loss of drug sensitivity is via the NF-kappaB2 pathway. PMID- 16260625 TI - Identification and characterization of Elf1, a conserved transcription elongation factor in Saccharomyces cerevisiae. AB - In order to identify previously unknown transcription elongation factors, a genetic screen was carried out to identify mutations that cause lethality when combined with mutations in the genes encoding the elongation factors TFIIS and Spt6. This screen identified a mutation in YKL160W, hereafter named ELF1 (elongation factor 1). Further analysis identified synthetic lethality between an elf1Delta mutation and mutations in genes encoding several known elongation factors, including Spt4, Spt5, Spt6, and members of the Paf1 complex. Genome-wide synthetic lethality studies confirmed that elf1Delta specifically interacts with mutations in genes affecting transcription elongation. Chromatin immunoprecipitation experiments show that Elf1 is cotranscriptionally recruited over actively transcribed regions and that this association is partially dependent on Spt4 and Spt6. Analysis of elf1Delta mutants suggests a role for this factor in maintaining proper chromatin structure in regions of active transcription. Finally, purification of Elf1 suggests an association with casein kinase II, previously implicated in roles in transcription. Together, these results suggest an important role for Elf1 in the regulation of transcription elongation. PMID- 16260626 TI - RelB/p52 NF-kappaB complexes rescue an early delay in mammary gland development in transgenic mice with targeted superrepressor IkappaB-alpha expression and promote carcinogenesis of the mammary gland. AB - Classical NF-kappaB (p65/p50) transcription factors display dynamic induction in the mammary gland during pregnancy. To further elucidate the role of NF-kappaB factors in breast development, we generated a transgenic mouse expressing the IkappaB-alpha S32/36A superrepressor (SR) protein under control of the mouse mammary tumor virus (MMTV) long terminal repeat promoter. A transient delay in mammary ductal branching was observed in MMTV-SR-IkappaB-alpha mice early during pregnancy at day 5.5 (d5.5) and d7.5; however, development recovered by mid- to late pregnancy (d14.5). Recovery correlated with induction of nuclear cyclin D1 and RelB/p52 NF-kappaB complexes. RelB/p52 complexes induced cyclin D1 and c-myc promoter activities and failed in electrophoretic mobility shift assay to interact with IkappaB-alpha-glutathione S-transferase, indicating that their weak interaction with IkappaB-alpha can account for the observed recovery of mammary gland development. Activation of IKKalpha and NF-kappaB-inducing kinase was detected by d5.5, implicating the alternative NF-kappaB signaling pathway in RelB/p52 induction. Constitutively active IKKalpha induced p52, RelB, and cyclin D1 in untransformed mammary epithelial cells. Moreover, mouse mammary tumors induced by 7,12-dimethylbenz(a)anthracene treatment displayed increased RelB/p52 activity. Inhibition of RelB in breast cancer cells repressed cyclin D1 and c-Myc levels and growth in soft agar. These results implicate RelB/p52 complexes in mammary gland development and carcinogenesis. PMID- 16260627 TI - Chromatin immunoprecipitation-based screen to identify functional genomic binding sites for sequence-specific transactivators. AB - In various human diseases, altered gene expression patterns are often the result of deregulated gene-specific transcription factor activity. To further understand disease on a molecular basis, the comprehensive analysis of transcription factor signaling networks is required. We developed an experimental approach, combining chromatin immunoprecipitation (ChIP) with a yeast-based assay, to screen the genome for transcription factor binding sites that link to transcriptionally regulated target genes. We used the tumor suppressor p53 to demonstrate the effectiveness of the method. Using primary and immortalized, nontransformed cultures of human mammary epithelial cells, we isolated over 100 genomic DNA fragments that contain novel p53 binding sites. This approach led to the identification and validation of novel p53 target genes involved in diverse signaling pathways, including growth factor signaling, protein kinase/phosphatase signaling, and RNA binding. Our results yield a more complete understanding of p53-regulated signaling pathways, and this approach could be applied to any number of transcription factors to further elucidate complex transcriptional networks. PMID- 16260628 TI - Enhanceosome formation over the beta interferon promoter underlies a remote control mechanism mediated by YY1 and YY2. AB - The expression of beta interferon genes from humans and mice is under the immediate control of a virus-responsive element (VRE) that terminates 110 bp upstream from the transcriptional start site. Whereas a wealth of information is available for the enhanceosome that is formed on the VRE upon the signals generated by viral infection, early observations indicating the existence of other far-upstream control elements have so far remained without a molecular fundament. Guided by a computational analysis of DNA structures, we could locate three as-yet-unknown transcription factor-binding regions at -0.5, -2, and -3 kb. Our present study delineates the interplay of factors YY1 and YY2 as it occurs at the sites at -3 kb and -2 kb (otherwise called HS1 and HS2), consistent with the idea that the novel factor YY2 antagonizes the negative actions exerted by YY1. Differences between the human and murine control regions will be described. PMID- 16260629 TI - Pendrin is a novel in vivo downstream target gene of the TTF-1/Nkx-2.1 homeodomain transcription factor in differentiated thyroid cells. AB - Thyroid transcription factor gene 1 (TTF-1) is a homeobox-containing gene involved in thyroid organogenesis. During early thyroid development, the homeobox gene Nkx-2.5 is expressed in thyroid precursor cells coincident with the appearance of TTF-1. The aim of this study was to investigate the molecular mechanisms underlying thyroid-specific gene expression. We show that the Nkx-2.5 C terminus interacts with the TTF-1 homeodomain and, moreover, that the expression of a dominant-negative Nkx-2.5 isoform (N188K) in thyroid cells reduces TTF-1-driven transcription by titrating TTF-1 away from its target DNA. This process reduced the expression of several thyroid-specific genes, including pendrin and thyroglobulin. Similarly, down-regulation of TTF-1 by RNA interference reduced the expression of both genes, whose promoters are sensitive to and directly associate with TTF-1 in the chromatin context. In conclusion, we demonstrate that pendrin and thyroglobulin are downstream targets in vivo of TTF 1, whose action is a prime factor in controlling thyroid differentiation in vivo. PMID- 16260630 TI - Trf4 and Trf5 proteins of Saccharomyces cerevisiae exhibit poly(A) RNA polymerase activity but no DNA polymerase activity. AB - The Saccharomyces cerevisiae Trf4 and Trf5 proteins are members of a distinct family of eukaryotic DNA polymerase beta-like nucleotidyltransferases, and a template-dependent DNA polymerase activity has been reported for Trf4. To define the nucleotidyltransferase activities associated with Trf4 and Tr5, we purified these proteins from yeast cells and show that whereas both proteins exhibit a robust poly(A) polymerase activity, neither of them shows any evidence of a DNA polymerase activity. The poly(A) polymerase activity, as determined for Trf4, is strictly Mn2+ dependent and highly ATP specific, incorporating AMP onto the free 3'-hydroxyl end of an RNA primer. Unlike the related poly(A) polymerases from other eukaryotes, which are located in the cytoplasm and regulate the stability and translation efficiency of specific mRNAs, the Trf4 and Trf5 proteins are nuclear, and a multiprotein complex associated with Trf4 has been recently shown to polyadenylate a variety of misfolded or inappropriately expressed RNAs which activate their degradation by the exosome. To account for the effects of Trf4/Trf5 proteins on the various aspects of DNA metabolism, including chromosome condensation, DNA replication, and sister chromatid cohesion, we suggest an additional and essential role for the Trf4 and Trf5 protein complexes in generating functional mRNA poly(A) tails in the nucleus. PMID- 16260631 TI - Mitochondrial lipid abnormality and electron transport chain impairment in mice lacking alpha-synuclein. AB - The presynaptic protein alpha-synuclein, implicated in Parkinson disease (PD), binds phospholipids and has a role in brain fatty acid (FA) metabolism. In mice lacking alpha-synuclein (Snca-/-), total brain steady-state mass of the mitochondria-specific phospholipid, cardiolipin, is reduced 22% and its acyl side chains show a 51% increase in saturated FAs and a 25% reduction in essential n-6, but not n-3, polyunsaturated FAs. Additionally, 23% reduction in phosphatidylglycerol content, the immediate biosynthetic precursor of cardiolipin, was observed without alterations in the content of other brain phospholipids. Consistent with these changes, more ordered lipid head group and acyl chain packing with enhanced rotational motion of diphenylhexatriene (DPH) about its long axis were demonstrated in time-resolved DPH fluorescence lifetime experiments. These abnormalities in mitochondrial membrane properties were associated with a 15% reduction in linked complex I/III activity of the electron transport chain, without reductions in mitochondrial number, complex II/III activity, or individual complex I, II, III, or IV activity. Reduced complex I activity is thought to be a critical factor in the development of PD. Thus, altered membrane composition and structure and impaired complex I/III function in Snca-/- brain suggest a relationship between alpha-synuclein's role in brain lipid metabolism, mitochondrial function, and PD. PMID- 16260632 TI - Predictive value of fetal pulmonary venous flow patterns in identifying the need for atrial septoplasty in the newborn with hypoplastic left ventricle. AB - BACKGROUND: Pulmonary venous Doppler (PVD) flow patterns in the fetus with hypoplastic left heart syndrome (HLHS) have been correlated with restrictive interatrial communication or intact atrial septum (RAS) postnatally; however, the ability of PVD to identify the neonate requiring emergent atrial septoplasty (EAS) for severe left atrial hypertension and hypoxemia has not been critically evaluated. It was the purpose of this study to determine the predictive power of fetal PVD in identifying the need for EAS in newborns with HLHS and RAS. METHODS AND RESULTS: Forty-one patients with fetal PVD flow analysis and postnatally confirmed HLHS were studied. Pulsed-wave assessment of PVD flow included S-, D-, and A-wave velocity, time-velocity integral (VTI) of forward and reverse flow, and S/D velocity and forward/reverse VTI ratio. Neonatal EAS was used as the primary clinical outcome variable. Receiver operating characteristic curves were used to determine cutpoints at which PVD indices best predicted EAS. Cutpoints were evaluated for clinical accuracy and usefulness by use of Bayesian analysis. Eight of 41 subjects underwent EAS. Need for EAS was most accurately predicted by forward/reverse VTI ratio <5 (sensitivity, 0.88, 95% CI, 0.49 to 0.99; specificity, 0.97, 95% CI, 0.82 to 0.99), which, when present, increases the posttest likelihood of EAS to 74%, assuming a pretest prevalence of 10%. Accuracy and usefulness of other PVD indices were affected by false-positive results. CONCLUSIONS: In the fetus with HLHS, a PVD forward/reverse VTI ratio of <5 is the strongest predictor of the need for EAS in the newborn period. These observations should improve our ability to identify and expectantly manage the fetus with HLHS and RAS. PMID- 16260633 TI - Temporally controlled onset of dilated cardiomyopathy through disruption of the SRF gene in adult heart. AB - BACKGROUND: Serum response factor (SRF) is a cardiac transcription factor involved in cell growth and differentiation. We have shown, using the Cre/loxP system, that cardiac-specific disruption of SRF gene in the embryonic heart results in lethal cardiac defects. The role of SRF in adult heart is unknown. METHODS AND RESULTS: We disrupted SRF in the adult heart using a heart-specific tamoxifen-inducible Cre recombinase. This disruption led to impaired left ventricular function with reduced contractility, subsequently progressing to dilated cardiomyopathy, as demonstrated by serial echocardiography, including tissue Doppler imaging. The cytoarchitecture of cardiomyocytes was altered in the intercalated disks. All mutant mice died from heart failure 10 weeks after treatment. These functional and structural defects were preceded by early alterations in the cardiac gene expression program: major decreases in mRNA levels for cardiac alpha-actin, muscle creatine kinase, and calcium-handling genes. CONCLUSIONS: SRF is crucial for adult cardiac function and integrity. We suggest that the rapid progression to heart failure in SRF mutant mice results primarily from decreased expression of proteins involved in force generation and transmission, low levels of polymerized actin, and changes in cytoarchitecture, without hypertrophic compensation. These cardiac-specific SRF-deficient mice have the morphological and clinical features of acquired dilated cardiomyopathy in humans and may therefore be used as an inducible model of this disorder. PMID- 16260634 TI - Reverse remodeling of the left cardiac chambers after catheter ablation after 1 year in a series of patients with isolated atrial fibrillation. AB - BACKGROUND: Isolated atrial fibrillation (AF) is associated with mild enlargement of the left atrium (LA) and left ventricular (LV) diastolic dysfunction. The impact of ablation of isolated AF on left chamber size and function is unclear, and whether diastolic dysfunction is the cause or the consequence of AF remains unknown. The objective of this prospective study was to evaluate the impact of sinus rhythm restoration by catheter ablation on LV diastolic dysfunction, LA morphology, and mechanical function. METHODS AND RESULTS: Forty-eight patients with isolated AF were studied by serial echocardiographic studies at baseline and at 1-, 3-, 6-, 9-, and 12-month intervals after radiofrequency ablation. LA dimensions and mechanical function and LV systolic and diastolic functions were evaluated at each time interval. Diastolic function was assessed with conventional Doppler parameters and new indexes such as tissue Doppler imaging, mitral flow propagation velocity, and combined criteria. LV diastolic dysfunction was present in paroxysmal and chronic AF patients with a reduction of tissue Doppler imaging lateral early diastolic peak velocity in 37% (P<0.001) and 48% (P<0.01), respectively, compared with healthy control subjects. At the end of the follow-up, LA area decreased significantly by 18% (P<0.001) in paroxysmal and 23% (P<0.05) in chronic AF patients. Diastolic function improved significantly with an increase in lateral early diastolic peak velocity of 29% (P<0.001) in paroxysmal AF and 46% (P<0.05) in chronic AF patients. A significant increase in LV ejection fraction was also noted for both groups: 7.7% and 18.8%, respectively. CONCLUSIONS: This study demonstrates reverse morphological remodeling of the LA and improvement of LV diastolic and systolic functions after restoration of sinus rhythm by ablation for isolated AF. Because patients with isolated AF have none of the traditional causes of LV diastolic dysfunction, our findings suggest that AF may be partly the cause rather than the consequence of diastolic dysfunction. PMID- 16260636 TI - The thromboxane A2 receptor antagonist S18886 prevents enhanced atherogenesis caused by diabetes mellitus. AB - BACKGROUND: S18886 is an orally active thromboxane A2 (TXA2) receptor (TP) antagonist in clinical development for use in secondary prevention of thrombotic events in cardiovascular disease. We previously showed that S18886 inhibits atherosclerosis in apolipoprotein E-deficient (apoE(-/-)) mice by a mechanism independent of platelet-derived TXA2. Atherosclerosis is accelerated by diabetes and is associated with increased TXA(2) and other eicosanoids that stimulate TP. The purpose of this study was to determine whether S18886 lessens the enhanced atherogenesis in diabetic apoE(-/-) mice. METHODS AND RESULTS: Diabetes mellitus was induced in apoE(-/-) mice with streptozotocin and was treated or not with S18886 (5 mg.kg(-1).d(-1)). After 6 weeks, aortic lesion area was increased >4 fold by diabetes in apoE(-/-) mice, associated with similar increases in serum glucose and cholesterol. S18886 largely prevented the diabetes-related increase in lesion area without affecting the hyperglycemia or hypercholesterolemia. S18886 prevented deterioration of endothelial function and endothelial nitric oxide synthase expression, as well as increases in intimal markers of inflammation associated with diabetes. In human aortic endothelial cells in culture, S18886 also prevented the induction of vascular cell adhesion molecule-1 and prevented the decrease in endothelial nitric oxide synthase expression caused by high glucose. CONCLUSIONS: The TP antagonist inhibits inflammation and accelerated atherogenesis caused by diabetes, most likely by counteracting effects on endothelial function and adhesion molecule expression of eicosanoids stimulated by the diabetic milieu. PMID- 16260635 TI - Decreased perivascular fibrosis but not cardiac hypertrophy in ROCK1+/- haploinsufficient mice. AB - BACKGROUND: Rho GTPase and its downstream target, Rho-associated kinase (ROCK), have been implicated in diverse cardiovascular diseases such as cardiac hypertrophy. However, pharmacological inhibitors of ROCK are not entirely specific, nor can they discriminate between the ROCK isoforms ROCK1 and ROCK2. To determine the specific role of ROCK1 in the development of cardiac hypertrophy, we generated ROCK1(+/-) haploinsufficient mice and determined whether cardiac hypertrophy and remodeling are decreased in these mice. METHODS AND RESULTS: Litters of ROCK1(-/-) mice on C57Bl/6 background were markedly underrepresented, suggesting lethality in utero or postnatally. ROCK1(+/-) mice, however, are viable and fertile with no obvious phenotypic abnormalities. Basal blood pressure, heart rate, and cardiac dimension and function in ROCK1(+/-) mice were similar to those in wild-type (WT) littermates. Infusion of angiotensin II (400 ng.kg(-1).min(-1) for 28 days) or treatment with NG-nitro-L-arginine methyl ester (1 mg/mL in drinking water for 28 days) caused similar increases in systolic blood pressure, left ventricular wall thickness, left ventricular mass, ratio of heart weight to tibial length, and cardiomyocyte size in ROCK1(+/-) mice and WT littermates. In contrast, perivascular fibrosis in hearts was increased to a lesser extent in ROCK1(+/-) mice compared with WT littermates. This was associated with decreased expression of transforming growth factor-beta, connective tissue growth factor, and type III collagen. In addition, perivascular fibrosis induced by transaortic constriction or myocardial infarction was decreased in ROCK1(+/-) mice compared with WT littermates. CONCLUSIONS: These findings indicate ROCK1 is critical for the development of cardiac fibrosis, but not hypertrophy, in response to various pathological conditions and suggest that signaling pathways leading to the hypertrophic and profibrotic response of the heart are distinct. PMID- 16260637 TI - Leukocyte engagement of platelet glycoprotein Ibalpha via the integrin Mac-1 is critical for the biological response to vascular injury. AB - BACKGROUND: Leukocyte-platelet interactions are critical in the initiation and progression of atherosclerosis as well as restenosis. Although the leukocyte integrin Mac-1 (alphaMbeta2, CD11b/CD18) has been implicated in the firm adhesion and transmigration of leukocytes at sites of platelet deposition, the precise alphaMbeta2 counterligand responsible for mediating adhesion-strengthening interactions between neutrophils and platelets in vivo has not previously been identified. METHODS AND RESULTS: Our previous studies have established the P201 K217 sequence in the alphaMI domain as the binding site for platelet glycoprotein (GP) Ibalpha. Here we report that antibody targeting of alphaM(P201-K217) reduced alphaMbeta2-dependent adhesion to GP Ibalpha but not other alphaMbeta2 ligands, including fibrinogen, intercellular adhesion molecule-1, and junctional adhesion molecule-3. Anti-alphaM(P201-K217) inhibited the firm adhesion of both human and murine leukocytes to adherent platelets under laminar flow conditions. In a mouse femoral artery wire injury model, antibody targeting of alphaM(P201-K217) reduced leukocyte accumulation after injury that was accompanied by inhibition of cellular proliferation and neointimal thickening. CONCLUSIONS: This study demonstrates that GP Ibalpha is a physiologically relevant ligand for alphaMbeta2 and that integrin engagement of GP Ibalpha is critical to leukocyte function and the biological response to vascular injury. These observations establish a molecular target for selectively disrupting leukocyte-platelet complexes that promote inflammation in thrombosis and restenosis. PMID- 16260638 TI - Chronic atrioventricular nodal vagal stimulation: first evidence for long-term ventricular rate control in canine atrial fibrillation model. AB - BACKGROUND: We have previously demonstrated that selective atrioventricular nodal (AVN) vagal stimulation (AVN-VS) can be used to control ventricular rate during atrial fibrillation (AF) in acute experiments. However, it is not known whether this approach could provide a long-term treatment in conscious animals. Thus, this study reports the first observations on the long-term efficacy and safety of this novel approach to control ventricular rate during AF in chronically instrumented dogs. METHODS AND RESULTS: In 18 dogs, custom-made bipolar patch electrodes were sutured to the epicardial AVN fat pad for delivery of selective AVN-VS by a subcutaneously implanted nerve stimulator (pulse width 100 micros or 1 ms, frequency 20 or 160 Hz, amplitude 6 to 10 V). Fast-rate right atrial pacing (600 bpm) was used to induce and maintain AF. ECG, blood pressure, and body temperature were monitored telemetrically. One week after the induction of AF, AVN-VS was delivered and maintained for at least 5 weeks. It was found that AVN VS had a consistent effect on ventricular rate slowing (on average 45+/-13 bpm) over the entire period of observation. Echocardiography showed improvement of cardiac indices with ventricular rate slowing. AVN-VS was well tolerated by the animals, causing no signs of distress or discomfort. CONCLUSIONS: Beneficial long term ventricular rate slowing during AF can be achieved by implantation of a nerve stimulator attached to the epicardial AVN fat pad. This novel concept is an attractive alternative to other methods of rate control and may be applicable in a selected group of patients. PMID- 16260639 TI - Absorption, metabolization, and antiplatelet effects of 300-, 600-, and 900-mg loading doses of clopidogrel: results of the ISAR-CHOICE (Intracoronary Stenting and Antithrombotic Regimen: Choose Between 3 High Oral Doses for Immediate Clopidogrel Effect) Trial. AB - BACKGROUND: For patients undergoing percutaneous coronary intervention, the administration of a clopidogrel loading dose ranging from 300 to 600 mg is currently recommended. It is unknown, though, whether loading doses higher than 600 mg exert additional suppression of platelet function. METHODS AND RESULTS: Sixty patients with suspected or documented coronary artery disease admitted to our hospital for coronary angiography were included in this trial. They were allocated to 1 of 3 clopidogrel loading doses (300, 600, or 900 mg) in a double blinded, randomized manner. Plasma concentrations of the active thiol metabolite, unchanged clopidogrel, and the inactive carboxyl metabolite of clopidogrel were determined before and serially after drug administration. Optical aggregometry was performed before and 4 hours after administration of clopidogrel. Loading with 600 mg resulted in higher plasma concentrations of the active metabolite, clopidogrel, and the carboxyl metabolite compared with loading with 300 mg (P< or =0.03) and lower values for adenosine diphosphate-induced (5 and 20 micromol/L) platelet aggregation 4 hours after drug administration (P=0.01 and 0.004). With administration of 900 mg, no further increase in plasma concentrations of active metabolite and clopidogrel (P> or =0.38) and no further suppression of adenosine diphosphate-induced (5 and 20 micromol/L) platelet aggregation 4 hours after drug administration was achieved when compared with administration of 600 mg (P=0.59 and 0.39). CONCLUSIONS: Single doses of clopidogrel higher than 600 mg are not associated with an additional significant suppression of platelet function because of limited clopidogrel absorption. PMID- 16260640 TI - Ghrelin improves endothelial function in patients with metabolic syndrome. AB - BACKGROUND: Metabolic syndrome importantly accelerates the atherosclerotic process, the earliest event of which is endothelial dysfunction. Ghrelin, a gastric peptide with cardiovascular actions, has been shown to inhibit proatherogenic changes in experimental models. This study therefore investigated whether ghrelin administration might beneficially affect endothelial function in metabolic syndrome. METHODS AND RESULTS: Endothelium-dependent and -independent vasodilator responses to intra-arterial infusion of increasing doses of acetylcholine and sodium nitroprusside (SNP), respectively, were assessed by strain-gauge plethysmography before and after local administration of human ghrelin (200 microg/min). During saline, the vasodilator response to acetylcholine was significantly blunted (P=0.008) in patients with metabolic syndrome (n=12, 5 female) compared with controls (n=12, 7 female), whereas the vasodilator response to SNP was not different between groups (P=0.68). In patients with metabolic syndrome, basal plasma ghrelin was significantly lower than in controls (P=0.02). In these patients, ghrelin infusion markedly increased intravascular concentrations of the peptide (P<0.001) and resulted in a potentiation of the vasodilator response to acetylcholine (P=0.001 versus saline) but not to SNP (P=0.22). This effect was likely related to enhanced nitric oxide bioavailability because, in a group of patients with metabolic syndrome (n=6, 2 female), ghrelin had no effect on the vasodilator response to acetylcholine (P=0.78 versus saline) after nitric oxide inhibition by NG-monomethyl-L-arginine. CONCLUSIONS: These findings indicate that ghrelin reverses endothelial dysfunction in patients with metabolic syndrome by increasing nitric oxide bioactivity, thereby suggesting that decreased circulating levels of the peptide, such as those found in these patients, might play a role in the pathobiology of atherosclerosis. PMID- 16260642 TI - Roadmaps, translational research, and childish curiosity. PMID- 16260641 TI - Angiotensin II type 1 receptor agonistic antibodies reflect fundamental alterations in the uteroplacental vasculature. AB - Abnormal uterine perfusion detected by Doppler sonography reflects impaired trophoblast invasion, a factor involved in the pathogenesis of pregnancy complications such as preeclampsia or intrauterine growth retardation. Recent studies have demonstrated an autoantibody against the angiotensin type 1 (AT1) receptor in pregnant women with preeclampsia. Our aim was to determine whether the AT1 autoantibody precedes the clinical symptoms and is thus predictive of preeclampsia. We therefore detected this antibody in serum from second trimester pregnancies with abnormal uterine perfusion because these women show an indirect sign of inadequate trophoblast invasion. Then the AT1 autoantibody distribution/concentration was compared with that of women at term with or without pregnancy pathology. The AT1 autoantibody was already detectable in second trimester pregnant women with abnormal uterine perfusion before the clinical manifestation of preeclampsia (80%). However, it was also found in second trimester pregnant women with abnormal uterine perfusion who later developed intrauterine growth retardation (60%) or even had a normal course of pregnancy (62%). In the third trimester, the AT1 autoantibody was demonstrated in 89% of patients with manifest preeclampsia, 86% of those with manifest intrauterine growth retardation, and even in healthy pregnant women at term with a history of abnormal uterine perfusion in the second trimester. We conclude that the AT1 autoantibody is an early but nonspecific marker for preeclampsia. The generation of this antibody seems to be associated with distinct types of pregnancy disorders resulting from impaired placental development. The AT1 autoantibody may thus be causative for pathological uteroplacental perfusion. PMID- 16260643 TI - Biological weapons and secrecy (WC 2300). PMID- 16260644 TI - Liposomes and the physico-chemical basis of unconsciousness. PMID- 16260646 TI - Dopamine receptors set the pattern of activity generated in subthalamic neurons. AB - Information processing in the brain requires adequate background neuronal activity. As Parkinson's disease progresses, patients typically become akinetic; the death of dopaminergic neurons leads to a dopamine-depleted state, which disrupts information processing related to movement in a brain area called the basal ganglia. Using agonists of dopamine receptors in the D1 and D2 families on rat brain slices, we show that dopamine receptors in these two families govern the firing pattern of neurons in the subthalamic nucleus, a crucial part of the basal ganglia. We propose a conceptual frame, based on specific properties of dopamine receptors, to account for the dominance of different background firing patterns in normal and dopamine-depleted states. PMID- 16260647 TI - Structure of infectious prions: stabilization by domain swapping. AB - A candidate structure for the minimal prion infectious unit is a recently discovered protein oligomer modeled as a beta-helical prion trimer (BPT); BPTs can stack to form cross-beta fibrils and may provide insight into protein aggregates of other amyloid diseases. However, the BPT lacks a clear intermonomer binding mechanism. Here we propose an alternative domain-swapped trimeric prion (DSTP) model and show with molecular dynamics (MD) that the DSTP has more favorable intermonomer hydrogen bonding and proline dihedral strain energy than the BPT. This new structural proposal may be tested by lysine and N terminus fluorescent resonance energy transfer (FRET) either directly on recombinant prion protein amyloid aggregates or on synthetic constructs that contain the proline/lysine-rich hinge region critical for domains to swap. In addition, the domain swapping may provide 1) intrinsic entanglement, which can contribute to the remarkable temperature stability of the infectious prion structure and help explain the absence of PrP(Sc) monomers, 2) insight into why specific prolines are potentially relevant to three inherited forms of prion disease, and 3) a simple explanation of prion strains assuming the strain is encoded in the monomer number of the oligomers. PMID- 16260648 TI - "Simple but not simpler": toward a unified picture of energy requirements in cell death. AB - In 1996, Wang and his group empirically disclosed a key role of (deoxy)-ATP in functioning of the apoptotic machinery. After almost a decade, and despite the emerged intricacy of the death pathways, ATP is still considered a key determinant of apoptosis with no apparent active roles in necrosis. Yet recent findings indicate that apoptosis proceeds even without energy and that necrosis can be regulated by ATP-dependent processes. This review strictly focuses on current knowledge on the role of energy in execution of different death programs. A thorough understanding of energy requirements in cell death can help to overcome obsolete dogmas in cell biology, paving the way to a more integrated, albeit not simpler, view of the molecular mechanisms contributing to cell dismantling. PMID- 16260649 TI - Establishment and characterization of renal progenitor like cells from S3 segment of nephron in rat adult kidney. AB - Kidney is thought to be a regenerative organ in terms of repair from acute tubular injury. It is unknown whether cell population contributes to repair disordered kidney. We attempted to identify and isolate highly proliferative cells from a single cell. We dissected a single nephron from adult rat kidney. Isolated nephrons were separated into segments and cultured. Outgrowing cells were replated after limiting dilution so that each well contained a single cell. One of cell line which was the most potent to grow was designated as rKS56. rKS56 cells showed cobblestone appearance and expressed immature cell markers relating to kidney development and mature tubular cell markers. rKS56 cells grew exponentially and could be maintained for 300 days without transformation. In different culture conditions, rKS56 cells differentiated into mature tubular cells defined by aquaporin-1, 2 expression, and responsiveness to parathyroid hormone or vasopressin. Engrafted to kidney in rat ischemic reperfusion model, rKS56 cells replaced in injured tubules in part after implantation and improved renal function. These results suggest rKS56 cells possess character such as self renewal, multi-plasticity and capability of tissue repair. rKS56 may possibly contribute to the future development of cell therapy for renal regeneration. PMID- 16260650 TI - Fibronectin is required for integrin alphavbeta6-mediated activation of latent TGF-beta complexes containing LTBP-1. AB - Transforming growth factor-betas (TGF-beta) are secreted as latent complexes consisting of the TGF-beta dimer, the TGF-beta propeptide dimer, and the latent TGF-beta binding protein (LTBP). Although the bonds between TGF-beta and its propeptide are cleaved intracellulary, the propeptide associates with TGF-beta by electrostatic interactions, thereby conferring latency to the complex. We reported that a specific sequence of LTBP-1 is required for latent TGF-beta activation by the integrin alphavbeta6. Here we describe a 24 amino acid sequence from the hinge domain required for activation. The LTBP-1 polypeptide rL1N, which includes the hinge, associates with fibronectin in binding assays. We present evidence that fibronectin null cells minimally activate latent TGF-beta and poorly incorporate the active hinge sequence into their matrix. In addition, cells missing the fibronectin receptor alpha5beta1 exhibit defective activation of latent TGF-beta by alphavbeta6 and decreased matrix incorporation. The results indicate specificity for integrin-mediated latent TGF-beta activation that include unique sequences in LTBP-1 and an appropriate matrix molecule. PMID- 16260651 TI - Characterization of vascular protein expression patterns in cerebral ischemia/reperfusion using laser capture microdissection and ICAT-nanoLC-MS/MS. AB - Cerebral ischemia rapidly initiates structural and functional changes in brain vessels, including blood-brain barrier disruption, inflammation, and angiogenesis. Molecular events that accompany these changes were investigated in brain microvessels extracted using laser-capture microdissection (LCM) from Sprague-Dawley rats subjected to a 20 min transient global cerebral ischemia followed by 1, 6, or 24 h reperfusion. Proteins extracted from approximately 300 LCM captured microvessels (20-100 microm) were ICAT-labeled and analyzed by nanoLC-MS. In-house software was used to identify paired ICAT peaks, which were then sequenced by nanoLC-MS/MS. Pattern analyses using k-means clustering method classified 57 differentially expressed proteins in 7 distinct dynamic patterns. Protein function was assigned using Panther Classification system. Early reperfusion (1 h) was characterized by down-regulation of ion pumps, nutrient transporters, and cell structure/motility proteins, and up-regulation of transcription factors, signal transduction molecules and proteins involved in carbohydrate metabolism. The up-regulation of inflammatory cytokines and proteins involved in the extracellular matrix remodeling and anti-oxidative defense was observed in late reperfusion (6-24 h). The up-regulation of IL-1beta and TGF 1beta in ischemic brain vessels was confirmed by ELISA, quantitative PCR, and/or immunohistochemistry. A biphasic postischemic (1 and 24 h) BBB opening for (3)H sucrose was evident in the same model. Differentially expressed proteins identified in brain vessels during reperfusion are likely involved in orchestrating functional vascular responses to ischemia, including the observed BBB disruption. PMID- 16260652 TI - SGLT-1-mediated glucose uptake protects intestinal epithelial cells against LPS induced apoptosis and barrier defects: a novel cellular rescue mechanism? AB - Excessive apoptosis induced by enteric microbes leads to epithelial barrier defects. This mechanism has been implicated in the pathogenesis of inflammatory bowel diseases (IBD) and bacterial enteritis. The sodium-dependent glucose cotransporter (SGLT-1) is responsible for active glucose uptake in enterocytes. The aim was to investigate the effects of SGLT-1 glucose uptake on enterocyte apoptosis and barrier defects induced by bacterial lipopolysaccharide (LPS). SGLT 1-transfected Caco-2 cells were treated with LPS (50 mug/mL) in low (5 mM) or high (25 mM) glucose media. LPS in low glucose induced caspase-3 cleavage, DNA fragmentation, and increased paracellular permeability to dextran in epithelial cells. These phenomena were significantly attenuated in high glucose. LPS increased SGLT-1 activity in high, but not low glucose media. Addition of phloridzin, which competitively binds to SGLT-1, inhibited the cytoprotection mediated by high glucose. Western blot showed that LPS in high glucose increased the levels of anti-apoptotic Bcl-2 and Bcl-X(L,) and did not change proapoptotic Bax. Differential extraction of membranous vs. cytosolic cell components demonstrated that high glucose inhibits mitochondrial cytochrome c translocation to cytosol. Collectively, SGLT-1-mediated glucose uptake increases anti-apoptotic proteins, and protects enterocytes from LPS-induced apoptosis and barrier defects. The understanding of this novel glucose-mediated rescue mechanism may lead to therapeutic interventions for various enteric diseases. PMID- 16260653 TI - Dual function of focal adhesion kinase in regulating integrin-induced MMP-2 and MMP-9 release by human T lymphoid cells. PMID- 16260654 TI - Ethics in orthopaedic surgery. PMID- 16260655 TI - Which research is to be believed? The ethics of industrial funding of orthopaedic research. PMID- 16260656 TI - Spinal infections. PMID- 16260657 TI - Current concepts in the management of femoroacetabular impingement. PMID- 16260658 TI - The puzzles of Perthes' disease: definitive studies of causal factors are needed. PMID- 16260659 TI - Sciatic nerve palsy secondary to postoperative haematoma in primary total hip replacement. AB - Sciatic nerve palsy is a recognised complication of primary total hip replacement. In our unit this complication was rare with an incidence of < 0.2% in the past ten years. We describe six cases of sciatic nerve palsy occurring in 355 consecutive primary total hip replacements (incidence 1.69%). Each of these palsies was caused by post-operative haematoma in the region of the sciatic nerve. Cases, which were recognised early and surgically-evacuated promptly, showed earlier and more complete recovery. Those patients for whom the diagnosis was delayed, and who were therefore managed expectantly, showed little or no recovery. Unexpected pain and significant swelling in the buttock, as well as signs of sciatic nerve irritation, suggest the presence of haematoma in the region of the sciatic nerve. It is, therefore, of prime importance to be vigilant for the features of a sciatic nerve palsy in the early post-operative period as, when recognised and treated early, the injury to the sciatic nerve may be reversed. PMID- 16260660 TI - The effect of hip resurfacing on oxygen concentration in the femoral head. AB - We inserted an electrode up the femoral neck into the femoral head of ten patients undergoing a metal-on-metal hip resurfacing arthroplasty through a posterior surgical approach and measured the oxygen concentration during the operation. In every patient the blood flow was compromised during surgery, but the extent varied. In three patients, the oxygen concentration was zero at the end of the procedure. The surgical approach caused a mean 60% drop (p < 0.005) in oxygen concentration while component insertion led to a further 20% drop (p < 0.04). The oxygen concentration did not improve significantly on wound closure. This study demonstrates that during hip resurfacing arthroplasty, patients experience some compromise to their femoral head blood supply and some have complete disruption. PMID- 16260661 TI - Mid-term wear characteristics of an uncemented acetabular component. AB - We investigated the rate of polyethylene wear of a cementless acetabular component at different periods of follow-up in order to test the hypothesis than an irrecoverable deformation process (creep) was followed by an initially low, but gradually increasing wear rate. We studied prospectively 93 uncemented total hip arthroplasties in 83 patients (mean age 50 years (22 to 63)) with a mean follow-up of 8.2 years (3 to 12). We measured the penetration of the femoral head from radiographs taken immediately after surgery at three, six and nine years, or at the latest follow-up. The median wear rate was 0.17 mm per year in the first three years, a finding which we considered to be caused by creep. Thereafter, the rate of wear declined to 0.07 mm per year (four- to six-year period) and then increased to 0.17 mm per year (seven to nine years) and 0.27 mm per year (more than nine years), which we considered to be a reflection of genuine polyethylene wear. After the nine-year follow-up the wear rates were higher in patients with marked osteolysis. We found no relationship between the inclination angle of the acetabular component or femoral head orientation and the rate of wear. No acetabular component required revision. PMID- 16260662 TI - Blood loss after total knee replacement: effects of computer-assisted surgery. AB - We carried out a prospective randomised study to evaluate the blood loss in 60 patients having a total knee arthroplasty and divided randomly into two equal groups, one having a computer-assisted procedure and the other a standard operation. The surgery was carried out by a single surgeon at one institution using a uniform approach. The only variable in the groups was the use of intramedullary femoral and tibial alignment jigs in the standard group and single tracker pins of the imageless navigation system in the tibia and femur in the navigated group. The mean drainage of blood was 1351 ml (715 to 2890; 95% confidence interval (CI) 1183 to 1518) in the computer-aided group and 1747 ml (1100 to 3030; CI 1581 to 1912) in the conventional group. This difference was statistically significant (p = 0.001). The mean calculated loss of haemoglobin was 36 g/dl in the navigated group versus 53 g/dl in the conventional group; this was significant at p < 0.00001. There was a highly significant reduction in blood drainage and the calculated Hb loss between the computer-assisted and the conventional techniques. This allows the ordering of less blood before the operation, reduces risks at transfusion and gives financial saving. Computer assisted surgery may also be useful for patients in whom blood products are not acceptable. PMID- 16260663 TI - Assessment of osteoarthritis after reconstruction of the anterior cruciate ligament: a study using single-photon emission computed tomography at ten years. AB - We used single-photon emission computed tomography (SPECT) to determine the long term risk of degenerative change after reconstruction of the anterior cruciate ligament (ACL). Our study population was a prospective series of 31 patients with a mean age at injury of 27.8 years (18 to 47) and a mean follow-up of ten years (9 to 13) after bone-patellar tendon-bone reconstruction of the ACL. The contralateral normal knee was used as a control. All knees were clinically stable with high clinical scores (mean Lysholm score, 93; mean Tegner activity score, 6). Fifteen patients had undergone a partial meniscectomy and ACL reconstruction at or before reconstruction of their ACL. In the group with an intact meniscus, clinical symptoms of osteoarthritis (OA) were found in only one patient (7%), who was also the only patient with marked isotope uptake on the SPECT scan compatible with OA. In the group which underwent a partial meniscectomy, clinical symptoms of OA were found in two patients (13%), who were among five (31%) with isotope uptake compatible with OA. Only one patient (7%) in this group had evidence of advanced OA on plain radiographs. The risk of developing OA after ACL reconstruction in this series is very low and lower than published figures for untreated ACL-deficient knees. There is a significant increase (p < 0.05) in degenerative change in patients who had a reconstruction of their ACL and a partial meniscectomy compared with those who had a reconstruction of their ACL alone. PMID- 16260664 TI - Oxford medial unicompartmental knee arthroplasty in patients younger and older than 60 years of age. AB - We present a comparison of the results of the Oxford unicompartmental knee arthroplasty in patients younger and older than 60 years of age. The ten-year all cause survival of the < 60 years of age group (52) was 91% (95% confidence interval (CI) 12), while in the > or = 60 years of age group (512), the figure was 96% (95% CI 3). For the younger group, the mean Hospital for Special Surgery score at ten-year follow-up (n = 21) was 94 of 100, compared with a mean of 86 of 100 for the older group (n = 135). The results show that the Oxford unicompartmental arthroplasty can achieve ten-year results that are comparable to total knee arthroplasty in patients < 60 years of age. We conclude that for patients aged over 50, age should not be considered a contraindication for this procedure. PMID- 16260665 TI - Ten-year in vivo wear measurement of a fully congruent mobile bearing unicompartmental knee arthroplasty. AB - Polyethylene particulate wear debris continues to be implicated in the aetiology of aseptic loosening following knee arthroplasty. The Oxford unicompartmental knee arthroplasty employs a spherical femoral component and a fully congruous meniscal bearing to increase contact area and theoretically reduce the potential for polyethylene wear. This study measures the in vivo ten-year linear wear of the device, using a roentgenstereophotogrammetric technique. In this in vivo study, seven medial Oxford unicompartmental prostheses, which had been implanted ten years previously were studied. Stereo pairs of radiographs were acquired for each patient and the films were analysed using a roentgen stereophotogrammetric analysis calibration and a computer-aided design model silhouette-fitting technique. Penetration of the femoral component into the original volume of the bearing was our estimate of linear wear. In addition, eight control patients were examined less than three weeks post-insertion of an Oxford prosthesis, where no wear would be expected. The control group showed no measured wear and suggested a system accuracy of 0.1 mm. At ten years, the mean linear wear rate was 0.02 mm/year. The results from this in vivo study confirm that the device has low ten year linear wear in clinical practice. This may offer the device a survival advantage in the long term. PMID- 16260666 TI - Men and women have similar Q angles: a clinical and trigonometric evaluation. AB - The Q angle is an important determinant of patellar tracking, though its clinical relevance is debatable. One controversy centres around any possible differences in its value between men and women. The accepted, though unproven explanation, for the greater Q angle in women is that a woman has a wider pelvis. However, because of the long distance between the pelvis and patella, relative to the distance from the patella to the tibial tuberosity, large changes in the position of the anterior superior iliac spine are necessary to effect significant changes in the Q angle. In our study of 69 subjects, we did not find such large differences in the position of the anterior superior iliac spine, and found a mean difference of only 2.3 degrees between the Q angles of men and women. Furthermore, we found that men and women of equal height demonstrated similar Q angles, with taller people having slightly smaller Q angles. The slight difference in Q angles between men and women can be explained by the fact that men tend to be taller. PMID- 16260667 TI - Osteolysis in a surface-cemented, primary, modular Freeman-Samuelson total knee replacement. AB - We analysed at a mean follow-up of 7.25 years the clinical and radiological outcome of 117 patients (125 knees) who had undergone a primary, cemented, modular Freeman-Samuelson total knee replacement. While the tibial and femoral components were cemented, the patellar component was uncemented. A surface cementing technique was used to secure the tibial components. A total of 82 knees was available for radiological assessment. Radiolucent lines were seen in 41 knees (50%) and osteolytic lesions were seen in 13 knees (16%). Asymptomatic, rotational loosening of the patellar implant was seen in four patients and osteolysis was more common in patients with a patellar resurfacing. Functional outcome scores were available for 41 patients (41 knees, 35%) and the mean Western Ontario McMasters Universities score was 77.5 (sd 19.5) and the cumulative survival was 93.4% at ten years with revision for aseptic loosening as an endpoint. Increased polyethylene wear from modular components, a rotationally loose patella, and the surface-cementing technique may have contributed to the high rate of osteolysis seen in our study. PMID- 16260668 TI - The Weil osteotomy: a seven-year follow-up. AB - We prospectively evaluated the one- and seven-year results of the Weil osteotomy for the treatment of metatarsalgia with subluxed or dislocated metatarsophalangeal joints in 25 feet of 24 patients. Good to excellent results were achieved in 21 feet (84%) after one year and in 22 (88%) after seven years. The American Orthopaedic Foot and Ankle Society score significantly improved from 48 (SD 15) points before surgery to 75 (SD 24) at one year, and 83 (SD 18) at seven years. The procedure significantly reduced pain, diminished isolated plantar callus formation and increased the patient's capacity for walking. Redislocation of the metatarsophalangeal joint was seen in two feet (8%) after one year and in three (12%) after seven years. Although floating toes and restricted movement of the metatarsophalangeal joint may occur, the Weil osteotomy is safe and effective. PMID- 16260669 TI - Tenodesis of the extensor carpi ulnaris for chronic, post-traumatic lunotriquetral instability. AB - We describe a technique of soft-tissue reconstruction which is effective for the treatment of chronic lunotriquetral instability. Part of extensor carpi ulnaris is harvested with its distal attachment preserved. It is passed through two drill holes in the triquetrum and sutured to itself. This stabilises the ulnar side of the wrist. We have reviewed 46 patients who underwent this procedure for post traumatic lunotriquetral instability with clinical signs suggestive of ulnar sided carpal instability. Standard radiographs were normal. All patients had pre operative arthroscopy of the wrist at which dynamic lunotriquetral instability was demonstrated. A clinical rating system for the wrist by the Mayo clinic was used to measure the outcome. In 19 patients the result was excellent, in ten good, in 11 satisfactory and in six poor. On questioning, 40 (87%) patients said that surgery had substantially improved the condition and that they would recommend the operation. However, six (13%) were unhappy with the outcome and would not undergo the procedure again for a similar problem. There were six complications, five of which related to pisotriquetral problems. The mean follow up was 39.1 months (6 to 100). We believe that tenodesis of extensor carpi ulnaris is a very satisfactory procedure for isolated, chronic post-traumatic lunotriquetral instability in selected patients. In those with associated pathology, the symptoms were improved, but the results were less predictable. PMID- 16260670 TI - The validity of the mangled extremity severity score in the assessment of upper limb injuries. AB - The Mangled Extremity Severity Score (MESS) may be used to decide whether to perform amputation in patients with injuries involving a limb. A score of 7 points or higher indicates the need for amputation. We have treated three patients with a MESS of 7 points or higher, in two of which the injured limb was salvaged. This scoring system was originally devised to assess injuries to the lower limb. However, a MESS of 7 points as a justification for amputation does not appear appropriate when assessing injuries to the major vessels in the upper limb. PMID- 16260671 TI - The use of a virtual three-dimensional model to evaluate the intraosseous space available for percutaneous screw fixation of acetabular fractures. AB - We created virtual three-dimensional reconstruction models from computed tomography scans obtained from patients with acetabular fractures. Virtual cylindrical implants were placed intraosseously in the anterior column, the posterior column and across the dome of the acetabulum. The maximum diameter which was entirely contained within the bone was determined for each position of the screw. In the same model, the cross-sectional diameters of the columns were measured and compared to the maximum diameter of the corresponding virtual implant. We found that the mean maximum diameter of virtual implant accommodated by the anterior columns was 6.4 mm and that the smallest diameter of the columns was larger than the maximum diameter of the equivalent virtual implant. This study suggests that the size of the screw used for percutaneous fixation of acetabular fractures should not be based solely on the measurement of cross sectional diameter and that virtual three-dimensional reconstructions might be useful in pre-operative planning. PMID- 16260672 TI - Incidence of infection with the use of non-irradiated morcellised allograft bone washed at the time of revision arthroplasty of the hip. AB - Implantation of allograft bone is an integral part of revision surgery of the hip. One major concern with its use is the risk of transmission of infective agents. There are a number of methods of processing allograft bone in order to reduce this risk. One method requires washing the tissue using pulsed irrigation immediately before implantation. We report the incidence of deep bacterial infection in 138 patients (144 revision hip arthroplasties) who had undergone implantation of allograft bone. The bone used was fresh-frozen, non-irradiated and pulse-washed with normal saline before implantation. The deep infection rate at a minimum follow-up of one year was 0.7%. This method of processing appears to be associated with a very low risk of allograft-related bacterial infection. PMID- 16260673 TI - Surgical treatment and outcome of conventional pelvic chondrosarcoma. AB - We reviewed 124 patients with a conventional pelvic chondrosarcoma who had been treated over a period of 20 years. We recorded the type of tumour (central or peripheral), type of operation (limb salvage surgery or hemipelvectomy), the grade of tumour, local recurrence and/or metastases, in order to identify the factors which might influence survival. More satisfactory surgical margins were achieved for central tumours or in those patients treated by hemipelvectomy. However, grade 1 tumours, whatever the course, did not develop metastases or cause death, while grade 3 tumours had the worst outcome and prognosis. Central, high-grade tumours require aggressive surgical treatment in order to achieve adequate surgical margins, particularly in those lesions located close to the sacroiliac joint. By contrast, grade 1 peripheral chondrosarcomas may be treated with contaminated margins in order to reduce operative morbidity, but without reducing survival. PMID- 16260674 TI - The incidence of Perthes' disease in Southwest Scotland. AB - We have investigated the annual incidence of Perthes' disease in Dumfries and Galloway (Southwest Scotland), in relation to the population density and socio economic status. The incidence of Perthes' disease in rural Scotland is comparable with that in urban areas (15.4 per 100,000). There was a direct association between the incidence of Perthes' disease and deprivation scores, with the highest incidence in the most deprived areas. A higher incidence of Perthes' disease was noted in areas with a lower population density compared with those with a higher density. We found no correlation between population density and deprivation scores for individual electoral wards within the region. PMID- 16260675 TI - A profile of Perthes' disease in Greater Glasgow: is there an association with deprivation? AB - It has been reported that there is an association between Perthes' disease and poverty. We examined the demographic data of a group of 240 children (263 hips) who presented with Perthes' disease in Greater Glasgow, where the mean deprivation scores are substantially greater than in the rest of Scotland, to see if this association applied and whether other clues to the aetiology of Perthes' disease could be found. There were 197 boys and 43 girls; 39 (16.25%) had a family history of Perthes' disease. Bone age in this series was heavily skewed towards the lower percentiles. The mean number of siblings was 1.9, with 31 (12.9%) being an only child. Maternal age at the birth of the first child showed no preponderance of older mothers. Maternal smoking during and after pregnancy was noted in 132 (55%), which compared with the 52% reported in the population of Greater Glasgow in general. Of the children in our series, 60 (25%) were in social class IV and V. However, this applies to more than half of the population of Greater Glasgow. There was no significant evidence of a preponderance of Perthes' disease in the most deprived groups. The aetiology of Perthes' disease is likely to be multifactorial and may include a genetic or deprivation influence resulting in delayed bone age. PMID- 16260676 TI - Operative treatment for hip subluxation in spinal muscular atrophy. AB - Subluxation of the hip is common in patients with intermediate spinal muscular atrophy. This retrospective study aimed to investigate the influence of surgery on pain and function, as well as the natural history of subluxed hips which were treated conservatively. Thirty patients were assessed clinically and radiologically. Of the nine who underwent surgery only one reported satisfaction and four had recurrent subluxation. Of the 21 patients who had no surgery, 18 had subluxation at the latest follow-up, but only one reported pain in the hip. We conclude that surgery for subluxation of the hip in these patients is not justified. PMID- 16260677 TI - Clinical features and microbiology in 204 cases of septic arthritis in Malawian children. AB - We examined 204 children (137 boys and 67 girls) aged 12 years and under with septic arthritis. Their mean age was 31.1 months (1 to 144; SD 41.6). The most common joints affected were the knees and shoulders. Joints in the upper limb were affected more often in younger children and in the lower limb in those who were older. The mean age for an infection was 12 months in the shoulder and 73 months in the hip. The most common organisms cultured were species of Salmonella. PMID- 16260678 TI - Haemarthroses after total knee arthroplasty caused by an isolated platelet factor 3 availability defect. AB - We present seven patients with recurrent haemarthroses after total knee arthroplasty, caused by an inherent platelet function defect. These patients developed painful knee swelling, persistent bleeding and/or wound breakdown, a platelet factor 3 availability defect being identified in all cases. Surgical exploration, with joint debridement, lavage and synovectomy, was performed in four patients who did not improve with conservative therapy. Histopathological examination of synovium revealed a focal synovial reaction with histiocytic infiltration, and occasional foreign-body giant cells. One patient required an early revision because of aseptic loosening of their tibial component. The condition was treated by single-donor platelet transfusions with good results. The diagnosis, management, and relevance of this disorder are discussed. PMID- 16260679 TI - Intraneural metastasis of a synovial sarcoma to a peripheral nerve. AB - We describe a case of intraneural metastasis of a synovial sarcoma, the first published case of a metastasis of a soft-tissue sarcoma to a peripheral nerve. PMID- 16260680 TI - A completely shattered tibia. AB - Despite advances in reconstructive surgery, salvage of mangled extremities still requires long periods of treatment with many operations that can be taxing both to the surgeon and the patient. Attempts at reconstruction of severely shattered limbs necessitate counselling with regard to the protracted course of treatment and associated morbidity as well as problems which may require abandoning of the procedure and secondary amputation. We report the successful salvage of a severely comminuted and open fracture of the tibia in a 32-year-old man. PMID- 16260681 TI - Finite-element analysis of failure of the Capital Hip designs. AB - The Capital Hip implant was a Charnley-based system which included a flanged and a roundback stem, both of which were available in stainless steel and titanium. The system was withdrawn from the market because of its inferior performance. However, all four of the designs did not produce poor rates of survival. Using a simulated-based, finite-element analysis, we have analysed the Capital Hip system. Our aim was to investigate whether our simulation was able to detect differences which could account for the varying survival between the Capital Hip designs, thereby further validating the simulation. We created finite-element models of reconstructions with the flanged and roundback Capital Hips. A loading history was applied representing normal walking and stair-climbing, while we monitored the formation of fatigue cracks in the cement. Corresponding to the clinical findings, our simulation was able to detect the negative effects of the titanium material and the flanged design in the Capital Hip system. Although improvements could be made by including the effect of the roughness of the surface of the stem, our study increased the value of the model as a predictive tool for determining failure of an implant. PMID- 16260682 TI - Iontophoresis as a means of delivering antibiotics into allograft bone. AB - Allograft bone is widely used in orthopaedic surgery, but peri-operative infection of the graft remains a common and disastrous complication. The efficacy of systemic prophylactic antibiotics is unproven, and since the graft is avascular it is likely that levels of antibiotic in the graft are low. Using an electrical potential to accelerate diffusion of antibiotics into allograft bone, high levels were achieved in specimens of both sheep and human allograft. In human bone these ranged from 187.1 mg/kg in endosteal (sd 15.7) to 124.6 (sd 46.2) in periosteal bone for gentamicin and 31.9 (sd 8.9) in endosteal and 2.9 (sd 1.1) in periosteal bone for flucloxacillin. The antibiotics remained active against bacteria in vitro after iontophoresis and continued to elute from the allograft for up to two weeks. Structural allograft can be supplemented directly with antibiotics using iontophoresis. The technique is simple and inexpensive and offers a potential means of reducing the rate of peri-operative infection in allograft surgery. Iontophoresis into allograft bone may also be applicable to other therapeutic compounds. PMID- 16260683 TI - Tissue restoration after resorption of polyglycolide and poly-laevo-lactic acid screws. AB - Despite worldwide clinical use of bio-absorbable devices for internal fixation in orthopaedic surgery, the degradation behaviour and tissue replacement of these implants are not fully understood. In a long-term experimental study, we have determined the patterns of tissue restoration 36 and 54 months after implantation of polyglycolic acid and poly-laevo-lactic acid screws in the distal femur of the rabbit. After 36 months in the polyglycolic acid group the specimens showed no remaining polymer and loose connective tissue occupied 80% of the screw track. Tissue restoration remained poor at 54 months, the amounts of trabecular bone and haematopoietic elements being significantly lower than those in the intact control group. The amount of trabecular bone within the screw track at 54 months in the polyglycolic acid group was less than in the empty drill holes (p = 0.04). In the poly-laevo-lactic acid group, polymeric material was present in abundance after 54 months, occupying 60% of the cross-section of the core area of the screw track. When using absorbable internal fixation implants we should recognise that the degradation of the devices will probably not be accompanied by the restoration of normal trabecular bone. PMID- 16260684 TI - Glove perforation and contamination in primary total hip arthroplasty. PMID- 16260685 TI - The complications of displaced intracapsular fractures of the hip. PMID- 16260686 TI - Glove perforation and contamination in primary total hip arthroplasty. PMID- 16260687 TI - Colorectal cancer surveillance: 2005 update of an American Society of Clinical Oncology practice guideline. AB - PURPOSE: To update the 2000 American Society of Clinical Oncology guideline on colorectal cancer surveillance. RECOMMENDATIONS: Based on results from three independently reported meta-analyses of randomized controlled trials that compared low-intensity and high-intensity programs of colorectal cancer surveillance, and on recent analyses of data from major clinical trials in colon and rectal cancer, the Panel recommends annual computed tomography (CT) of the chest and abdomen for 3 years after primary therapy for patients who are at higher risk of recurrence and who could be candidates for curative-intent surgery; pelvic CT scan for rectal cancer surveillance, especially for patients with several poor prognostic factors, including those who have not been treated with radiation; colonoscopy at 3 years after operative treatment, and, if results are normal, every 5 years thereafter; flexible proctosigmoidoscopy [corrected] every 6 months for 5 years for rectal cancer patients who have not been treated with pelvic radiation; history and physical examination every 3 to 6 months for the first 3 years, every 6 months during years 4 and 5, and subsequently at the discretion of the physician; and carcinoembryonic antigen every 3 months postoperatively for at least 3 years after diagnosis, if the patient is a candidate for surgery or systemic therapy. Chest x-rays, CBCs, and liver function tests are not recommended, and molecular or cellular markers should not influence the surveillance strategy based on available evidence. PMID- 16260688 TI - Getting melanoma cells to stimulate with frequency. PMID- 16260689 TI - Can statin therapy reduce the risk of breast cancer? PMID- 16260690 TI - The irreversibility of radiation-induced fibrosis: fact or folklore? PMID- 16260691 TI - Disease-free survival versus overall survival as a primary end point for adjuvant colon cancer studies: a commentary. PMID- 16260692 TI - Carboplatin for stage I seminoma and the sword of Damocles. PMID- 16260693 TI - Low-dose outpatient chemobiotherapy with temozolomide, granulocyte-macrophage colony stimulating factor, interferon-alpha2b, and recombinant interleukin-2 for the treatment of metastatic melanoma. AB - PURPOSE: The objective of this study was to further investigate the efficacy and safety of low-dose outpatient chemobiotherapy in patients with unresectable metastatic melanoma. PATIENTS AND METHODS: Thirty-one patients with histologically confirmed unresectable measurable metastatic melanoma were enrolled onto an open-label, multicenter phase II study. The treatment regimen consisted of oral temozolomide followed by subcutaneous biotherapy with granulocyte macrophage colony-stimulating factor, interferon-alfa, and recombinant interleukin-2 (rIL-2). RESULTS: Twenty-eight patients (90%) had M1c disease, and 58% had three or more sites of metastasis. Four patients (13%), all with M1c disease, had a complete response, and four patients had a partial response. The median progression-free survival was 4.9 months and the median overall survival was 13.1 months. Two patients (6%) developed CNS metastasis as the first site of disease progression, and 7 (23%) of 30 experienced CNS progression after receiving chemobiotherapy. A total of 112 cycles of therapy were administered. Toxicity occurred in 78% of the cycles and was grade 1 or 2 in the majority of cases and easily managed. Grade 4 toxicity occurred in 3% of the cycles. CONCLUSION: This low-dose chemobiotherapy combination produces clinical responses in patients with metastatic melanoma, even in those with M1c disease, is well tolerated, and allows home dosing. It offers a reasonable alternative to high-dose regimens, such as high-dose biochemotherapy or rIL-2 requiring prolonged periods of hospitalization, or single agent outpatient regimens, such as dacarbazine, which is usually not effective in patients with M1c disease. Furthermore, it may protect against the development of brain metastases. PMID- 16260694 TI - Use of statins and breast cancer: a meta-analysis of seven randomized clinical trials and nine observational studies. AB - PURPOSE: A growing body of evidence suggests that statins may have chemopreventive potential against breast cancer. Laboratory studies demonstrate that statins induce apoptosis and reduce cell invasiveness in various cell lines, including breast carcinoma cells. However, the clinical relevance of these data remains unclear. The nonconclusive nature of the epidemiologic data prompted us to conduct a detailed meta-analysis of the studies published on the subject in peer-reviewed literature. PATIENTS AND METHODS: A comprehensive search for articles published up until 2005 was performed; reviews of each study were conducted; and data were abstracted. Before meta-analysis, the studies were evaluated for publication bias and heterogeneity. Pooled relative risk (RR) estimates and 95% CIs were calculated using the random and the fixed-effects models. Subgroup and sensitivity analyses were also performed. RESULTS: Seven large randomized trials and nine observational studies (five case-control and four cohort studies) contributed to the analysis. We found no evidence of publication bias or heterogeneity among the studies. Statin use did not significantly affect breast cancer risk (fixed effects model: RR = 1.03; 95% CI, 0.93 to 1.14; random effects model: RR = 1.02; 95% CI, 0.89 to 1.18). When the analyses were stratified into subgroups, there was no evidence that study design substantially influenced the estimate of effects. Furthermore, the sensitivity analysis confirmed the stability of our results. CONCLUSION: Our meta-analysis findings do not support a protective effect of statins against breast cancer. However, this conclusion is limited by the relatively short follow-up times of the studies analyzed. Further studies are required to investigate the potential decrease in breast cancer risk among long-term statin users. PMID- 16260695 TI - Kinetics of response to long-term treatment combining pentoxifylline and tocopherol in patients with superficial radiation-induced fibrosis. AB - PURPOSE: Significant regression of radiation (RT) -induced fibrosis (RIF) has been achieved after treatment combining pentoxifylline (PTX) and alpha-tocopherol (vitE). In this study, we focus on the maximum response, how long it takes to achieve response, and changes after treatment discontinuation. PATIENTS AND METHODS: Measurable superficial RIF was assessed in patients treated by RT for breast cancer in a long-treatment (24 to 48 months) PTX-vitE (LPE) group of 37 patients (47 RIFs) and in a short-treatment (6 to 12 months) PTX-vitE (SPE) group of seven patients (eight RIFs). Between April 1995 and April 2000, women were treated with a daily combination of PTX (800 mg) and VitE (1,000 IU). RESULTS: Combined PTX-vitE was continuously effective and resulted in exponential RIF surface area regression (-46% for LPE and -68% for SPE at 6 months, -58% for LPE and -69% for SPE at 12 months, -63% for LPE and -62% for SPE at 18 months, and 68% for LPE at 24 and 36 months). The mean estimated maximal treatment effect was 68% RIF surface area regression. The mean time to this effect was 24 months and was shorter (16 months) in more recent RIF (< 6 years since RT) than in older RIF (28 months; P = .0003). Symptom severity (Subjective Objective Medical Management and Analytic Evaluation score) was halved in both groups. After treatment discontinuation, mean RIF surface area at 1 year had increased by +40% in the SPE group (rebound) and +8.5% in the LPE group. CONCLUSION: Under combined PTX-vitE treatment, RIF regression was exponential, with a two-thirds maximum response after a mean of 2 years. There was a risk of a rebound effect if treatment was too short. Long treatment (>/= 3 years) is recommended in patients with severe RIF. PMID- 16260696 TI - Immunogenicity, including vitiligo, and feasibility of vaccination with autologous GM-CSF-transduced tumor cells in metastatic melanoma patients. AB - PURPOSE: To determine the feasibility, toxicity, and immunologic effects of vaccination with autologous tumor cells retrovirally transduced with the GM-CSF gene, we performed a phase I/II vaccination study in stage IV metastatic melanoma patients. PATIENTS AND METHODS: Sixty-four patients were randomly assigned to receive three vaccinations of high-dose or low-dose tumor cells at 3-week intervals. Tumor cell vaccine preparation succeeded for 56 patients (88%), but because of progressive disease, the well-tolerated vaccination was completed in only 28 patients. We analyzed the priming of T cells against melanoma antigens, MART-1, tyrosinase, gp100, MAGE-A1, and MAGE-A3 using human leukocyte antigen/peptide tetramers and functional assays. RESULTS: The high-dose vaccination induced the infiltration of T cells into the tumor tissue. Three of 14 patients receiving the high-dose vaccine showed an increase in MART-1- or gp100-specific T cells in the peripheral blood during vaccination. Six patients experienced disease-free survival for more than 5 years, and two of these patients developed vitiligo at multiple sites after vaccination. MART-1- and gp100-specific T cells were found infiltrating in vitiligo skin. Upon vaccination, the T cells acquired an effector phenotype and produced interferon gamma on specific antigenic stimulation. CONCLUSION: We conclude that vaccination with GM-CSF-transduced autologous tumor cells has limited toxicity and can enhance T-cell activation against melanocyte differentiation antigens, which can lead to vitiligo. Whether the induction of autoimmune vitiligo may prolong disease-free survival of metastatic melanoma patients who are surgically rendered as having no evidence of disease before vaccination is worthy of further investigation. PMID- 16260697 TI - Temozolomide in combination with interferon-alfa versus temozolomide alone in patients with advanced metastatic melanoma: a randomized, phase III, multicenter study from the Dermatologic Cooperative Oncology Group. AB - PURPOSE: Temozolomide (TMZ) has shown efficacy in metastatic melanoma equal to that of dacarbazine (DTIC), the standard chemotherapeutic agent for melanoma. As the combination with interferon-alfa (IFN-alpha) appears superior to single-agent DTIC regarding response rates, the purpose of this study was to compare TMZ alone and TMZ plus IFN-alpha in terms of objective response (OR), overall survival, and safety in a prospective, randomized, multicenter trial. PATIENTS AND METHODS: Two hundred ninety-four patients with untreated stage IV metastatic melanoma (American Joint Committee on Cancer staging system) were randomly assigned to receive either oral TMZ alone (200 mg/m2/day; days 1 through 5 every 28 days) or in combination with subcutaneous IFN-alpha (5 MU/m2; days 1, 3, and 5 every week). RESULTS: Two hundred eighty-two patients were eligible for an intent-to treat analysis, 271 patients were treated per protocol. In the TMZ + IFN-alpha arm, 33 (24.1%) of 137 patients responded to therapy (partial or complete remission) whereas in the monotherapy arm, in 18 (13.4%) of 134 patients, a response was evident. Thus, the response rate was significantly higher in the combination arm (P = .036). Median survival time was 8.4 months for patients treated with TMZ (95% CI, 7.07 to 9.27) and 9.7 months for those treated with the combination (95% CI, 8.26 to 11.18; P = .16). Dose modifications and interval prolongations due to hematologic toxicity were significantly more frequent in the TMZ + IFN-alpha arm (P < .001). CONCLUSION: In metastatic melanoma treatment with TMZ + IFN-alpha leads to a significantly superior OR rate compared to treatment with TMZ alone, which did not translate into prolonged survival in our study population. PMID- 16260698 TI - Risk-adapted management for patients with clinical stage I seminoma: the Second Spanish Germ Cell Cancer Cooperative Group study. AB - PURPOSE: To assess the efficacy of a risk-adapted treatment policy for patients with stage I seminoma by using universally accepted risk criteria. PATIENTS AND METHODS: Between 1999 and 2003, 314 patients with clinical stage I seminoma after orchiectomy were prospectively included. One hundred patients (31.8%) presented no risk factors and were managed with surveillance. In contrast, 131 patients (41.7%) had tumors larger than 4 cm, 33 patients (10.5%) had rete testis involvement, and 50 patients (15.9%) had both risk factors. All the latter received two courses of adjuvant carboplatin. RESULTS: Chemotherapy was well tolerated, as only 17 patients (7.9%) presented grade 3 to 4 toxicity. Relapses were observed in six patients (6.0%) on surveillance and in seven patients (3.3%) treated with carboplatin (0.8% of tumors larger than 4 cm, 9.1% of those involving the rete testis, and 6.0% of patients with both risk criteria). All were located at the retroperitoneum, except for one at the spermatic cord. Median tumor size was 25 mm (range, 11 to 70 mm), and median time to relapse was 9 months (range, 4 to 28 months). All patients were rendered disease-free with chemotherapy (etoposide plus cisplatin). Median follow-up was 34 months (range, 12 to 72 months). The actuarial 5-year disease-free survival rate was 93.4% for patients on surveillance and 96.2% for patients treated with adjuvant chemotherapy. Overall 5-year survival was 100%. CONCLUSION: Adjuvant carboplatin is effective in reducing the relapse rate in patients with stage I seminoma and risk factors. A risk-adapted strategy is safe and feasible and should be considered an alternative to systematic approaches, such as irradiation, chemotherapy, or surveillance. PMID- 16260699 TI - Building upon the standard of care in adjuvant therapy of high-risk melanoma. PMID- 16260700 TI - Disease-free survival versus overall survival as a primary end point for adjuvant colon cancer studies: individual patient data from 20,898 patients on 18 randomized trials. AB - PURPOSE: A traditional end point for colon adjuvant clinical trials is overall survival (OS), with 5 years demonstrating adequate follow-up. A shorter-term end point providing convincing evidence to allow treatment comparisons could significantly speed the translation of advances into practice. METHODS: Individual patient data were pooled from 18 randomized phase III colon cancer adjuvant clinical trials. Trials included 43 arms, with a pooled sample size of 20,898 patients. The primary hypothesis was that disease-free survival (DFS), with 3 years of follow-up, is an appropriate primary end point to replace OS with 5 years of follow-up. RESULTS: The recurrence rates for years 1 through 5 were 12%, 14%, 8%, 5%, and 3%, respectively. Median time from recurrence to death was 12 months. Eighty percent of recurrences were in the first 3 years; 91% of patients with recurrence by 3 years died before 5 years. Correlation between 3 year DFS and 5-year OS was 0.89. Comparing control versus experimental arms within each trial, the correlation between hazard ratios for DFS and OS was 0.92. Within-trial log-rank testing using both DFS and OS provided the same conclusion in 23 (92%) of 25 cases. Formal measures of surrogacy were satisfied. CONCLUSION: In patients treated on phase III adjuvant colon clinical trials, DFS and OS are highly correlated, both within patients and across trials. These results suggest that DFS after 3 years of median follow-up is an appropriate end point for adjuvant colon cancer clinical trials of fluorouracil-based regimens, although marginally significant DFS improvements may not translate into significant OS benefits. PMID- 16260702 TI - Emotional health of black and white dementia caregivers: a contextual examination. AB - The aim of this study was to examine emotional health (e.g., depressive symptomatology) among 295 Black and 425 White dementia caregivers. I selected caregivers from the Resources for Enhancing Alzheimer's Caregivers' Health (REACH) data set. I used regression analysis to examine racial differences and the relationship between the contextual variables and depressive symptoms. Overall, age, size of social network, satisfaction with social activities, financial difficulty, positive aspects of caregiving, stressfulness appraisal, and health problems were related to depressive symptoms. Contextual factors were not homogeneous across race; the number of memory and behavior problems of the care recipient was associated with more reports of depressive symptoms among Black caregivers only. Results suggest the importance of examining various contexts surrounding the actual environment in which Black and White dementia caregivers provide care. PMID- 16260701 TI - Prospective, randomized, multicenter, double-blind placebo-controlled trial comparing adjuvant interferon alfa and isotretinoin with interferon alfa alone in stage IIA and IIB melanoma: European Cooperative Adjuvant Melanoma Treatment Study Group. AB - PURPOSE: The combination of interferon alfa (IFNalpha) and isotretinoin has shown a direct antiproliferative effect on human melanoma cell lines, but it remained unclear whether this combination is more effective than IFNalpha alone in patients with metastatic melanoma. We evaluated safety and efficacy of IFNalpha and isotretinoin compared with IFNalpha alone as adjuvant treatment in patients with primary malignant melanoma stage IIA and IIB. PATIENTS AND METHODS: In a prospective, randomized, double-blind, placebo-controlled trial, 407 melanoma patients in stage IIA (301 patients) and IIB (106 patients) were randomly assigned to either IFNalpha and isotretinoin (isotretinoin group; 206 patients) or IFNalpha and placebo (placebo group; 201 patients) after excision of the primary tumor. IFNalpha was administered three times a week at a dose of 3 million units subcutaneously for 24 months. Isotretinoin at a dose of 20 mg for patients < or = 73 kg, 30 mg for patients greater than 73 kg, or placebo daily for 24 months. RESULTS: A scheduled interim analysis revealed no significant differences in survival rates, with the isotretinoin group and the placebo group showing 5-year disease-free survival rates of 55% (95% CI, 46% to 65%) and 67% (95% CI, 59% to 75%), respectively, and overall 5-year survival rates of 76% (95% CI, 67% to 84%) and 81% (95% CI, 74% to 88%), respectively. The trial was stopped for futility. CONCLUSION: The addition of isotretinoin to an adjuvant treatment of low-dose IFNalpha in patients with stage IIA and IIB melanoma had no significant effect on disease-free or overall survival and is therefore not recommended. PMID- 16260703 TI - Longitudinal analysis of the reciprocal effects of self-assessed global health and depressive symptoms. AB - This study examined whether a reciprocal relationship exists between measures of self-assessed global health and depressive symptoms, net of covariates that included chronic illness, functional disability, education, income, gender, race, and age. Analyses of five waves of data from the Rand version of the Health and Retirement Survey (N=7,475), using an autoregressive, cross-lagged panel design, indicated that self-assessed overall health had a modest but statistically significant and consistent effect on depressive symptoms. In contrast, the level of depressive symptoms had a statistically nonsignificant effect on self-assessed health. There has been growing interest in identifying the factors that inform self-assessments of overall health. The present findings indicate that self assessed global health is not simply a manifestation of depressed affect. PMID- 16260704 TI - Understanding the relative importance of positive and negative social exchanges: examining specific domains and appraisals. AB - Negative social exchanges have been more reliably related to psychological health than have positive social exchanges. Little research, however, has sought to understand how underlying appraisal processes link such exchanges to psychological health. This study examined the frequencies of occurrence and appraisals of four parallel domains of positive and negative exchanges in relation to positive well-being and psychological distress in a national sample of 916 older adults. Structural equation analyses revealed that negative exchanges were related both to less well-being and greater psychological distress, whereas positive exchanges were related only to positive well-being. Furthermore, results supported a process in which appraisals mediated the link between social exchanges and psychological health. This social appraisal process helps explain the disproportionate impact of negative exchanges on psychological health. PMID- 16260705 TI - Depressive symptoms among African American and white older adults. AB - Guided by a stress and coping model, we explored determinants of depressive symptoms among community samples of older African Americans (n=255) and older Whites (n=452). We gave focus to the effects of demographic variables, physical health constraints (chronic conditions and functional disability), and psychosocial attributes (sense of mastery, religiosity, social support, and satisfaction with support), along with their interactive roles. We identified lower education, greater functional disability, lower sense of mastery, and poorer satisfaction with support as common risk factors for depressive symptoms in both groups; in contrast, the effects of age, gender, and religiosity were race specific. In addition, we obtained significant interactions among predictor variables in each group, identifying risk-reducing and risk-enhancing factors within each group. PMID- 16260706 TI - Trajectories of cognitive decline and social relations. AB - We assessed conjoint trajectories of cognitive decline and social relations over 7 years on a representative sample of community-dwelling elderly persons. We analyzed data using repeated measurement models. Social integration, family ties, and engagement with family were associated with cognitive function at advanced ages, controlling for education and introducing depressive symptoms, functional limitations, and chronic conditions as intervening variables. Association of social integration, through participation in community activities, with change in cognitive decline was more significant at advanced ages. Having friends was significantly associated with change in cognitive function in women only. Our findings have important implications for clinical medicine and public health because associations of social relations with cognitive function suggests that they may help to maintain cognitive function in old age. PMID- 16260707 TI - Influence of neuroticism, ethnicity, familism, and social support on perceived burden in dementia caregivers: pilot test of the transactional stress and social support model. AB - In this study we assessed the new transactional stress and social support model, postulating the role of neuroticism, ethnicity, familism, and social support in perceived burden in dementia caregivers. We used a convenience sample (N=77) of African American and White dementia caregivers. Results substantiated interrelationships among social support variables, and the influence of perceived positive social support on burden. Neuroticism was related to the perception of positive social support and burden. Results corroborated the model, focusing on neuroticism and quality of social support in modeling perceived burden in family caregivers. Findings call attention to the role of presumably long-standing individual differences in neuroticism that influence caregiver appraisals of stress and social support. PMID- 16260708 TI - Strength training in older adults: does desire determine confidence? AB - This study investigated conceptually relevant determinants of change in self efficacy beliefs related to muscular strength in a prospective study design. Results suggested that older adults' desire for muscular strength is an important determinant of changes in self-efficacy for strength. Studies involving older adults in strength training and physical rehabilitation should explore the development of behavioral interventions that target increases in such motives to be used in conjunction with these programs. PMID- 16260709 TI - Moderating role of marital quality in older adults' depressed affect: beyond the main-effects model. AB - We examine the role of three indicators of marital quality (marital disagreement, marital happiness, and time spent together) as moderators of the association between physical disability and depressed affect among married older individuals (N=1,044). We found support for the moderating role of marital disagreement wherein the detrimental effect of disability on depressed affect was significantly heightened among older adults with more disagreements with their spouse; a moderating effect was not detected for marital happiness or time spent together. We conclude that, in addition to its main effect on older adults' depressed affect, marital quality (as indicated by marital disagreement) plays a significant stress-moderating role in the physical disability-depressed affect link. PMID- 16260711 TI - On the shoulders of a giant: the legacy of Matilda White Riley for gerontology. AB - The legacy of Matilda White Riley to the study of aging encompasses multiple disciplines and extends to multiple domains within these disciplines. Although her greatest intellectual legacy is in sociology, she presented a compelling vision of the need for other disciplines to consider the role of social forces in shaping both aging as an individual, lifelong process and age as a feature of culture and social systems. This article reviews Riley's theoretical contributions in four areas: (1) articulating age and social systems; (2) identifying fallacies in the interpretation of research on aging; (3) theorizing about social change and structural lag; and (4) presenting social possibilities related to age integration. We conclude by considering briefly the reach of her legacy beyond sociology-in collaborating across disciplinary boundaries, in encouraging the development of sound longitudinal data archives, and in developing an institutional infrastructure to support and sustain research on aging in the social and behavioral sciences. Although short of physical stature, Riley's contributions to gerontology are enormous. Gerontologists from many disciplinary backgrounds have been informed by and rely upon these insights and thus share the advantage of "standing on the shoulders of a giant." PMID- 16260712 TI - Class, cohort, and consumption: the British experience of the third age. AB - OBJECTIVE: This study examines the relative effect of historical and social class location on engagement in consumerism within two different cohorts of retired people in the United Kingdom. METHODS: With use of self-reported data from the retired members of a nationally representative survey, an index of consumption was constructed. Its internal reliability was analysed and analyses of variance performed to examine the impact of class of origin, cohort, and class at exit on levels of self-reported "consumerism." RESULT: The index demonstrated sufficient internal reliability to provide an operational measurement of "consumerism" within this retired population. Analyses of variance confirmed that class at exit and cohort but not class of origin contributed significantly to variation in levels of consumerism. These effects were not mediated by cohort differences in health. DISCUSSION: Occupational standing immediately before retirement was associated with "consumerism" after retirement. This is unsurprising. The results also show that birth cohort exercised a stronger historical influence on current consumption in later life than did class background (i.e., paternal social class). This supports our thesis that the limited but increasing immersion in mass consumer society of successive cohorts who were born and grew up earlier in the 20th century continues to be reflected in levels of "consumerism" in retirement. PMID- 16260713 TI - Social networks among men and women: the effects of age and socioeconomic status. AB - OBJECTIVES: This study examines the main and interactive effects of age and socioeconomic status (SES) on social networks. METHODS: Respondents are drawn from a regional stratified probability sample aged 40 to 93 years. Hierarchical regression analysis estimates the influence of age and SES on dimensions of social networks, controlling for marital status and health among men and women. RESULTS: Among men, older age was associated with older networks. Professional men report networks that are less geographically proximal, however, occupational effects are most obvious in late life. Among women, age is associated with smaller networks that are older, less geographically proximal, and less frequently contacted. Whereas less education is associated with younger network members in midlife, among women in later life, lower levels of education are not associated with a younger network. Professional women report older networks composed of a higher proportion of friends than do homemakers. Higher levels of education are linked to larger personal networks among men and women, but not to the number of individuals considered closest. Among women, higher levels of education are also associated with less proximal networks. DISCUSSION: An examination of within-group variability reveals influences of age and SES on personal networks among men and women. PMID- 16260714 TI - Proxy sources for information on nursing home residents' quality of life. AB - OBJECTIVES: This study explores how well staff and family proxies' reports on selected quality-of-life (QOL) domains (comfort, dignity, functional competence, privacy, meaningful activity, food enjoyment, relationships, security, and autonomy) correspond to residents' own reports. METHODS: We compared QOL domain scores for nursing home residents and 1,326 staff proxies and 989 family proxies at the individual and facility level using means, Pearson correlation statistics, and intraclass correlations. Regression models adjusted for residents' age, gender, length of stay, ability to perform activities of daily living, and cognition. RESULTS: For each domain in more than half the cases, proxy means were within 1 SD of the resident means. Resident and family proxy individual reports for selected domains were correlated at 0.14 to 0.46 (all p <.000). Resident and staff proxy individual reports were correlated at 0.13 to 0.37 (all p <.000). Correlation of mean levels by facility for staff proxies was 0.26 to 0.64 (generally p <.05) and for family proxies 0.13 to 0.61 (p <.01 except for one domain). DISCUSSION: Although staff and family proxy domain scores are significantly correlated with resident scores, the level of correlation suggests they cannot simply be substituted for resident reports of QOL. Determining how proxy reports can be used for residents who cannot be interviewed at all remains an unresolved challenge. PMID- 16260715 TI - A refined protocol for coding nursing home residents' comments during satisfaction interviews. AB - OBJECTIVE: This study's objective was to refine a method for coding nursing home (NH) residents' comments about their perceptions of care into unmet needs specific to the manner and frequency of care delivery. METHODS: NH residents (N=69) were interviewed with both closed-ended (i.e., forced-choice) and open ended (i.e., residents' own words) questions about their perceptions of care across eight care domains. Unmet needs included comments indicating that residents desired a change in staff- and non-staff-related care. Staff-related unmet needs were further coded into unmet emotional support (i.e., emotional support or manner of care delivery) and instrumental (i.e., instrumental support or frequency of care) needs. RESULT: Of 66 residents who commented, 66% expressed at least one unmet need across eight care domains. Among these 44 residents, 52% and 84% had unmet emotional support and instrumental support needs, respectively (kappa=68 and.92). An additional 18% expressed both unmet emotional support and instrumental support needs. DISCUSSION: The refined method offers a systematic way to code residents' comments about their care into unmet needs related to the manner and frequency of care delivery. The findings have direct implications for the identification of care areas in need of improvement from the resident's perspective and the evaluation of improvement efforts. PMID- 16260716 TI - Self-rated health in a longitudinal perspective: a 9-year follow-up twin study. AB - OBJECTIVES: This study first considers age and cohort explanations for age related changes in mean values and variance in self-rated health. Second, it evaluates the contributions of genes and environments to self-rated health measured longitudinally. METHODS: Subjects were participants in the Swedish Adoption/Twin Study of Aging. Self-rated health assessments were collected in four waves over a 9-year follow-up period, from one or both members of 788 twin pairs. Linear mixed effect models were used to test for differences in means and variances. Structural equation modeling provided estimates of genetic and environmental components of variance and contributions to stability. RESULTS: Changes in means and variance within cohorts seem to reflect illness. Earlier born cohorts are more variable and have lower self-rated health. These cohort differences were not explained by childhood socioeconomic status. Correlations between time points reflect both environmental and genetic factors. DISCUSSION: Both genes and environments contribute to self-rated health longitudinally, and both age and cohort effects are seen. Age-related changes in self-rated health can be attributed to illness. Cohort differences are most likely attributable to socially mediated and individual-specific environmental factors. PMID- 16260717 TI - Letter to the editor. "Putting a human face on gerontologic research: identity issues". PMID- 16260718 TI - Letter to the editor. Qualitative and quantitative research methods. PMID- 16260719 TI - Thyroid hormone receptor beta mutants: Dominant negative regulators of peroxisome proliferator-activated receptor gamma action. AB - Thyroid hormone (T3) and peroxisome proliferators have overlapping metabolic effects in the maintenance of lipid homeostasis. Their actions are mediated by their respective receptors: thyroid hormone receptors (TR) and peroxisome proliferator-activated receptors (PPAR). We recently found that a dominantly negative TRbeta mutant (PV) that causes a genetic disease, resistance to thyroid hormone, acts to repress the ligand (troglitazone)-mediated transcriptional activity of PPARgamma in cultured thyroid cells. This finding suggests that TRbeta mutants could crosstalk with PPARgamma-signaling pathways. The present study explored the molecular mechanisms by which PV represses the PPARgamma transcriptional activity. Gel-shift assays show that the PV, similar to wild-type TRbeta, bound to the peroxisome proliferator response element (PPRE) as homodimers and heterodimers with PPARgamma or the retinoid X receptor (RXR), thereby competing with PPARgamma for binding to PPRE and for sequestering RXR. Association of PPRE-bound PV with corepressors [e.g., nuclear receptor corepressor (NCoR)] that led to transcriptional repression was independent of T3 and troglitazone. Chromatin immunoprecipitation assay further demonstrated that, despite the presence of ligands, NCoR was recruited to PPRE-bound PV on a PPARgamma-target gene, the lipoprotein lipase, in vivo, suggesting the dominant action of PV on PPARgamma-mediated transcriptional activity. Thus, the dominant negative action of PV is not limited on the wild-type TRs. The findings that TRbeta mutants affect PPARgamma functions through dominant negative action provide insights into the molecular mechanisms by which TR regulates the PPARgamma-target genes involved in metabolic pathways, lipid homeostasis, and carcinogenesis. PMID- 16260720 TI - Epidermal growth factor receptor signaling is required for normal ovarian steroidogenesis and oocyte maturation. AB - The midcycle luteinizing hormone (LH) surge triggers several tightly linked ovarian processes, including steroidogenesis, oocyte maturation, and ovulation. We designed studies to determine whether epidermal growth factor receptor (EGFR) mediated signaling might serve as a common regulator of these activities. Our results showed that EGF promoted steroidogenesis in two different in vitro models of oocyte-granulosa cell complexes. Inhibition of the EGFR kinase prevented EGF induced steroidogenesis in these in vitro systems and blocked LH-induced steroidogenesis in intact follicles primed with pregnant mare serum gonadotropin. Similarly, inhibition of the EGFR kinase attenuated LH-induced steroidogenesis in MA-10 Leydig cells. Together, these results indicate that EGFR signaling is critical for normal gonadotropin-induced steroidogenesis in both male and female gonads. Interestingly, inhibition of metalloproteinase-mediated cleavage of membrane-bound EGF moieties abrogated LH-induced steroidogenesis in ovarian follicles but not MA-10 cells, suggesting that LH receptor signaling activates the EGFR by different mechanisms in these two models. Finally, steroids promoted oocyte maturation in several ovarian follicle models, doing so by signaling through classical steroid receptors. We present a model whereby steroid production may serve as one of many integrated signals triggered by EGFR signaling to promote oocyte maturation in gonadotropin-stimulated follicles. PMID- 16260721 TI - The architecture of pre-mRNAs affects mechanisms of splice-site pairing. AB - The exon/intron architecture of genes determines whether components of the spliceosome recognize splice sites across the intron or across the exon. Using in vitro splicing assays, we demonstrate that splice-site recognition across introns ceases when intron size is between 200 and 250 nucleotides. Beyond this threshold, splice sites are recognized across the exon. Splice-site recognition across the intron is significantly more efficient than splice-site recognition across the exon, resulting in enhanced inclusion of exons with weak splice sites. Thus, intron size can profoundly influence the likelihood that an exon is constitutively or alternatively spliced. An EST-based alternative-splicing database was used to determine whether the exon/intron architecture influences the probability of alternative splicing in the Drosophila and human genomes. Drosophila exons flanked by long introns display an up to 90-fold-higher probability of being alternatively spliced compared with exons flanked by two short introns, demonstrating that the exon/intron architecture in Drosophila is a major determinant in governing the frequency of alternative splicing. Exon skipping is also more likely to occur when exons are flanked by long introns in the human genome. Interestingly, experimental and computational analyses show that the length of the upstream intron is more influential in inducing alternative splicing than is the length of the downstream intron. We conclude that the size and location of the flanking introns control the mechanism of splice-site recognition and influence the frequency and the type of alternative splicing that a pre-mRNA transcript undergoes. PMID- 16260722 TI - Methylprednisolone neutralizes the beneficial effects of erythropoietin in experimental spinal cord injury. AB - Inflammation plays a major pathological role in spinal cord injury (SCI). Although antiinflammatory treatment using the glucocorticoid methyprednisolone sodium succinate (MPSS) improved outcomes in several multicenter clinical trials, additional clinical experience suggests that MPSS is only modestly beneficial in SCI and poses a risk for serious complications. Recent work has shown that erythropoietin (EPO) moderates CNS tissue injury, in part by reducing inflammation, limiting neuronal apoptosis, and restoring vascular autoregulation. We determined whether EPO and MPSS act synergistically in SCI. Using a rat model of contusive SCI, we compared the effects of EPO [500-5,000 units/kg of body weight (kg-bw)] with MPSS (30 mg/kg-bw) for proinflammatory cytokine production, histological damage, and motor function at 1 month after a compression injury. Although high-dose EPO and MPSS suppressed proinflammatory cytokines within the injured spinal cord, only EPO was associated with reduced microglial infiltration, attenuated scar formation, and sustained neurological improvement. Unexpectedly, coadministration of MPSS antagonized the protective effects of EPO, even though the EPO receptor was up-regulated normally after injury. These data illustrate that the suppression of proinflammatory cytokines alone does not necessarily prevent secondary injury and suggest that glucocorticoids should not be coadministered in clinical trials evaluating the use of EPO for treatment of SCI. PMID- 16260723 TI - Carbonic anhydrase IV and XIV knockout mice: roles of the respective carbonic anhydrases in buffering the extracellular space in brain. AB - Previous studies have implicated extracellular carbonic anhydrases (CAs) in buffering the alkaline pH shifts that accompany neuronal activity in the rat and mouse hippocampus. CAs IV and XIV both have been proposed to mediate this extracellular buffering. To examine the relative importance of these two isozymes in this and other physiological functions attributed to extracellular CAs, we produced CA IV and CA XIV knockout (KO) mice by targeted mutagenesis and the doubly deficient CA IV/XIV KO mice by intercrossing the individual null mice. Although CA IV and CA XIV null mice both are viable, the CA IV nulls are produced in smaller numbers than predicted, indicating either fetal or postnatal losses, which preferentially affect females. CA IV/XIV double KO mice are also produced in fewer numbers than predicted and are smaller than WT mice, and many females die prematurely before and after weaning. Electrophysiological studies on hippocampal slices on these KO mice showed that either CA can mediate buffering after synaptic transmission in hippocampal slices in the absence of the other, but that eliminating both is nearly as effective as the CA inhibitor, benzolamide, in blocking the buffering seen in the WT mice. Thus, both CA IV and CA XIV contribute to extracellular buffering in the central nervous system, although CA IV appears to be more important in the hippocampus. These individual and double KO mice should be valuable tools in clarifying the relative contributions of each CA to other physiological functions where extracellular CAs have been implicated. PMID- 16260725 TI - Generation of tissue-specific transgenic birds with lentiviral vectors. AB - Birds are of great interest for a variety of research purposes, and effective methods for manipulating the avian genome would greatly accelerate progress in fields that rely on birds as model systems for biological research, such as developmental biology and behavioral neurobiology. Here, we describe a simple and effective method for producing transgenic birds. We used lentiviral vectors to produce transgenic quails that express GFP driven by the human synapsin gene I promoter. Expression of GFP was specific to neurons and consistent across multiple generations. Expression was sufficient to allow visualization of individual axons and dendrites of neurons in vivo by intrinsic GFP fluorescence. Tissue-specific transgene expression at high levels provides a powerful tool for biological research and opens new avenues for genetic manipulation in birds. PMID- 16260724 TI - A cAMP-response element binding protein-induced microRNA regulates neuronal morphogenesis. AB - MicroRNAs (miRNAs) regulate cellular fate by controlling the stability or translation of mRNA transcripts. Although the spatial and temporal patterning of miRNA expression is tightly controlled, little is known about signals that induce their expression nor mechanisms of their transcriptional regulation. Furthermore, few miRNA targets have been validated experimentally. The miRNA, miR132, was identified through a genome-wide screen as a target of the transcription factor, cAMP-response element binding protein (CREB). miR132 is enriched in neurons and, like many neuronal CREB targets, is highly induced by neurotrophins. Expression of miR132 in cortical neurons induced neurite outgrowth. Conversely, inhibition of miR132 function attenuated neuronal outgrowth. We provide evidence that miR132 regulates neuronal morphogenesis by decreasing levels of the GTPase-activating protein, p250GAP. These data reveal that a CREB-regulated miRNA regulates neuronal morphogenesis by responding to extrinsic trophic cues. PMID- 16260726 TI - Detection of functional single-nucleotide polymorphisms that affect apoptosis. AB - Human EBV-transformed B lymphocyte cell lines (LCLs) were used to measure the apoptotic response of individuals to gamma radiation. The responses form a normal distribution around a median of 35.5% apoptosis with a range of 12-58%. This heterogeneous response has a genetic basis. LCLs from Caucasian donors and African American donors form distinct distributions of apoptotic response; all of the 11 LCLs comprising the lowest responding group (exhibiting between 12-20% apoptosis) are from Caucasian donors. The assay is capable of detecting significant effects of SNPs in two genes, MDM2 and AKT1, whose products are involved in controlling the p53 pathway and cellular response to DNA damage, suggesting that these data and this assay can be used to identify novel SNPs in other genes whose products impact the cellular response to radiation. Finally, the LCLs in the lowest apoptotic response group have the highest concentration of AKT1 protein and all harbor a haplotype in AKT1 that is present in Caucasians but absent in African Americans. PMID- 16260727 TI - Dental senescence in a long-lived primate links infant survival to rainfall. AB - Primates tend to be long-lived, and, except for humans, most primate females are able to reproduce into old age. Although aging in most mammals is accompanied by dental senescence due to advanced wear, primates have low-crowned teeth that wear down before old age. Because tooth wear alters crown features gradually, testing whether early dental senescence causes reproductive senescence has been difficult. To identify whether and when low-crowned teeth compromise reproductive success, we used a 20-year field study of Propithecus edwardsi, a rainforest lemur from Madagascar with a maximum lifespan of >27 years. We analyzed tooth wear in three dimensions with dental topographic analysis by using Geographical Information Systems (GIS) technology. We report that tooth wear exposes compensatory shearing blades that maintain dental function for 18 years. Beyond this age, female fertility remains high; however infants survive only if lactation seasons have elevated rainfall. Therefore, low-crowned teeth accommodate wear to a point, after which reproductive success closely tracks environmental fluctuations. These results suggest a tooth wear-determined, but rainfall-mediated, onset of reproductive senescence. Additionally, our study indicates that even subtle changes in climate may affect reproductive success of rainforest species. PMID- 16260728 TI - Fate of dietary perchlorate in lactating dairy cows: Relevance to animal health and levels in the milk supply. AB - Perchlorate is a goitrogenic anion that competitively inhibits the sodium iodide transporter and has been detected in forages and in commercial milk throughout the U.S. The fate of perchlorate and its effect on animal health were studied in lactating cows, ruminally infused with perchlorate for 5 weeks. Milk perchlorate levels were highly correlated with perchlorate intake, but milk iodine was unaffected, and there were no demonstrable health effects. We provide evidence that up to 80% of dietary perchlorate was metabolized, most likely in the rumen, which would provide cattle with a degree of refractoriness to perchlorate. Data presented are important for assessing the environmental impact on perchlorate concentrations in milk and potential for relevance to human health. PMID- 16260729 TI - An immune-responsive serpin, SRPN6, mediates mosquito defense against malaria parasites. AB - We have functionally analyzed the orthologous SRPN6 genes from Anopheles stephensi and Anopheles gambiae using phylogenetic, molecular, reverse genetic, and cell biological tools. The results strongly implicate SRPN6 in the innate immune response against Plasmodium. This gene belongs to a mosquito-specific gene cluster including three additional Anopheles serpins. SRPN6 expression is induced by Escherichia coli and both rodent and human malaria parasites. The gene is specifically expressed in midgut cells invaded by Plasmodium ookinetes and in circulating and attached hemocytes. Knockdown of SRPN6 expression by RNA interference in susceptible An. stephensi leads to substantially increased parasite numbers, whereas depletion in susceptible An. gambiae delays progression of parasite lysis without affecting the number of developing parasites. However, the An. gambiae SRPN6 knockdown increases the number of melanized parasites in the L3-5 refractory strain and in susceptible G3 mosquitoes depleted of CTL4. These results indicate that AsSRPN6 is involved in the parasite-killing process, whereas AgSRPN6 acts on parasite clearance by inhibiting melanization and/or promoting parasite lysis. We propose that these observed phenotypic differences are due to changed roles of the respective target serine proteases in the two mosquito species. PMID- 16260730 TI - Mullerian inhibiting substance acts as a motor neuron survival factor in vitro. AB - The survival of motor neurons is controlled by multiple factors that regulate different aspects of their physiology. The identification of these factors is important because of their relationship to motor neuron disease. We investigate here whether Mullerian Inhibiting Substance (MIS) is a motor neuron survival factor. We find that motor neurons from adult mice synthesize MIS and express its receptors, suggesting that mature motor neurons use MIS in an autocrine fashion or as a way to communicate with each other. MIS was observed to support the survival and differentiation of embryonic motor neurons in vitro. During development, male-specific MIS may have a hormone effect because the blood-brain barrier has yet to form, raising the possibility that MIS participates in generating sex-specific differences in motor neurons. PMID- 16260732 TI - Ocular dominance plasticity is stably maintained in the absence of alpha calcium calmodulin kinase II (alphaCaMKII) autophosphorylation. AB - The molecule alpha calcium calmodulin kinase II (alphaCaMKII) is known to play a fundamental role in the induction of many forms of synaptic plasticity. A major theory of alphaCaMKII function proposes that autophosphorylation of the molecule mediates not only the induction but also the maintenance of synaptic plasticity. To test this hypothesis, we assessed ocular dominance plasticity in genetically engineered mice that carry a mutation preventing autophosphorylation of alphaCaMKII. These mutant mice are deficient in plasticity after monocular deprivation, but a sufficiently long period of monocular deprivation will induce ocular dominance plasticity. After induction of ocular dominance plasticity, the stability of the induced changes was assayed after binocular deprivation. Plasticity in homozygous mutant animals was as stable as that measured in WT littermates; also, response characteristics did not differ between the two groups. Our results suggest that alphaCaMKII autophosphorylation is required for the induction of ocular dominance plasticity but is not needed for its stable maintenance thereafter. PMID- 16260731 TI - IL-32 synergizes with nucleotide oligomerization domain (NOD) 1 and NOD2 ligands for IL-1beta and IL-6 production through a caspase 1-dependent mechanism. AB - The activation of innate immunity requires the amplification of signals induced by pattern-recognition receptors for bacterial products. We have investigated the role of the newly described cytokine IL-32 in the amplification of cytokine production induced by the two most clinically relevant families of microbial receptors, the cell-surface Toll-like receptors (TLRs) and the intracellular nuclear oligomerization domain (NOD) receptor family. IL-32 synergized with the NOD1- and NOD2-specific muropeptides of peptidoglycans for the release of IL 1beta and IL-6 (a 3- to 10-fold increase). In contrast, IL-32 did not influence the cytokine production induced via TLRs. The synergistic effect of IL-32 and synthetic muramyl dipeptide (MDP) on cytokine production was absent in the cells of patients with Crohn's disease bearing the NOD2 frameshift mutation 3020insC, demonstrating that the IL-32/MDP synergism depends on NOD2. This in vitro synergism between IL-32 and NOD2 ligands was consistent with a marked constitutive expression of IL-32 in human colon epithelial tissue. In addition, the potentiating effect of IL-32 on the cytokine production induced by the synthetic muropeptide FK-156 was absent in NOD1-deficient macrophages, supporting the interaction between IL-32 and NOD1 pathways. When specific caspase inhibitors were used, the synergism between IL-32 and MDP/NOD2 depended on the activation of caspase 1. Only additive effects of IL-32 and muropeptides were observed for TNF alpha production. The modulation of intracellular NOD2 pathways by IL-32, but not cell-surface TLRs, and the marked expression of IL-32 in colon mucosa suggest a role of IL-32 in the pathogenesis of Crohn's disease. PMID- 16260733 TI - A persistent pesticide residue and the unusual catalytic proficiency of a dehalogenating enzyme. AB - The soil of potato fields in The Netherlands harbors bacteria with the ability to metabolize 3-chloroacrylic acid, generated by the degradation of a pesticide (1,3 dichloropropene) that entered the environment in 1946. From examination of rate constants at elevated temperatures, we infer that the half-time at 25 degrees C for spontaneous hydrolytic dechlorination of trans-3-chloroacrylic acid is 10,000 years, several orders of magnitude longer than half-times for spontaneous decomposition of other environmental pollutants such as 1,2-dichloroethane (72 years), paraoxon (13 months), atrazine (5 months), and aziridine (52 h). With thermodynamic parameters for activation similar to those for the spontaneous hydration of fumarate at pH 6.8, this slow reaction proceeds at a constant rate through the pH range between 2 and 12. However, at the active site of the enzyme 3-chloroacrylate dehalogenase (CaaD), isolated from a pseudomonad growing in these soils, hydrolytic dechlorination proceeds with a half-time of 0.18 s. Neither k(cat) nor k(cat)/K(m) is reduced by increasing solvent viscosity with trehalose, implying that the rate of enzymatic dechlorination is controlled by chemical events in catalysis rather than by diffusion-limited substrate binding or product release. CaaD achieves an approximately 10(12)-fold rate enhancement, matching or surpassing the rate enhancements produced by many enzymes that act on more conventional biological substrates. One of those enzymes is 4-oxalocrotonate tautomerase, with which CaaD seems to share a common evolutionary origin. PMID- 16260734 TI - Rapid perceptual integration of facial expression and emotional body language. AB - In our natural world, a face is usually encountered not as an isolated object but as an integrated part of a whole body. The face and the body both normally contribute in conveying the emotional state of the individual. Here we show that observers judging a facial expression are strongly influenced by emotional body language. Photographs of fearful and angry faces and bodies were used to create face-body compound images, with either matched or mismatched emotional expressions. When face and body convey conflicting emotional information, judgment of facial expression is hampered and becomes biased toward the emotion expressed by the body. Electrical brain activity was recorded from the scalp while subjects attended to the face and judged its emotional expression. An enhancement of the occipital P1 component as early as 115 ms after presentation onset points to the existence of a rapid neural mechanism sensitive to the degree of agreement between simultaneously presented facial and bodily emotional expressions, even when the latter are unattended. PMID- 16260735 TI - Studies on the regulation of ornithine decarboxylase in yeast: effect of deletion in the MEU1 gene. AB - Methylthioadenosine is formed during the biosynthesis of spermidine and of spermine and is metabolized by methylthioadenosine phosphorylase, an enzyme missing in several tumor cell lines. In Saccharomyces cerevisiae, this enzyme is coded by the MEU1 gene. We have now studied the effect of the meu1 deletion on polyamine metabolism in yeast. We found that the effects of the meu1Delta mutation mostly depend on the stage of cell growth. As the cell density increases, there is a marked fall in the level of ornithine decarboxylase (ODC) in the MEU1(+) cells, which we show is caused by an antizyme-requiring degradation system. In contrast, there is only a small decrease in the ODC level in the meu1Delta cells. The meu1Delta cells have a higher putrescine and a lower spermidine level than MEU1(+) cells, suggesting that the decreased spermidine level in the meu1Delta cultures is responsible for the greater apparent stability of ODC in the meu1Delta cells. The lower spermidine level in the meu1Delta cells probably results from an inhibition of spermidine synthase by the methylthioadenosine that presumably accumulates in these mutants. In both MEU1(+) and the meu1Delta cultures, the ODC levels were markedly decreased by the addition of spermidine to the media, and thus our results contradict the postulation of Subhi et al. [Subhi, A. L., et al. (2003) J. Biol. Chem. 278, 49868-49873] of a novel regulatory pathway in meu1Delta cells in which ODC is not responsive to spermidine. PMID- 16260737 TI - Evidence for capillarity contributions to gecko adhesion from single spatula nanomechanical measurements. AB - The hairy attachment system on a gecko's toes, consisting of one billion spatulae in the case of Gekko gecko [Ruibal, R. & Ernst, V. (1965) J. Morphol. 117, 271 294], allows it to adhere to nearly all surface topographies. The mechanistic basis for gecko adhesion has been intensely investigated, but the lowest hierarchical level, that of the spatula, has become experimentally accessible only recently. This report details measurements of the adhesion force exerted by a single gecko spatula for various atmospheric conditions and surface chemistries. Through judicious choice and modification of substrates, the short- and long-range adhesive forces are separated. In contrast to previous work [Autumn, K., Sitti, M., Liang, Y. C. A., Peattie, A. M., Hansen, W. R., Sponberg, S., Kenny, T. W., Fearing, R., Israelachvili, J. N. & Full, R. J. (2002) Proc. Natl. Acad. Sci. USA 99, 12252-12256], our measurements clearly show that humidity contributes significantly to gecko adhesion on a nanoscopic level. These findings are crucial for the development of artificial biomimetic attachment systems. PMID- 16260736 TI - Structural basis for the recognition between HIV-1 integrase and transcriptional coactivator p75. AB - Integrase (IN) is an essential retroviral enzyme, and human transcriptional coactivator p75, which is also referred to as lens epithelium-derived growth factor (LEDGF), is the dominant cellular binding partner of HIV-1 IN. Here, we report the crystal structure of the dimeric catalytic core domain of HIV-1 IN complexed to the IN-binding domain of LEDGF. Previously identified LEDGF hotspot residues anchor the protein to both monomers at the IN dimer interface. The principal structural features of IN that are recognized by the host factor are the backbone conformation of residues 168-171 from one monomer and a hydrophobic patch that is primarily comprised of alpha-helices 1 and 3 of the second IN monomer. Inspection of diverse retroviral primary and secondary sequence elements helps to explain the apparent lentiviral tropism of the LEDGF-IN interaction. Because the lethal phenotypes of HIV-1 mutant viruses unable to interact with LEDGF indicate that IN function is highly sensitive to perturbations of the structure around the LEDGF-binding site, we propose that small molecule inhibitors of the protein-protein interaction might similarly disrupt HIV-1 replication. PMID- 16260738 TI - Pharmacological induction of heat-shock proteins alleviates polyglutamine mediated motor neuron disease. AB - Spinal and bulbar muscular atrophy (SBMA) is an adult-onset motor neuron disease caused by the expansion of a trinucleotide CAG repeat encoding the polyglutamine tract in the first exon of the androgen receptor gene (AR). The pathogenic, polyglutamine-expanded AR protein accumulates in the cell nucleus in a ligand dependent manner and inhibits transcription by interfering with transcriptional factors and coactivators. Heat-shock proteins (HSPs) are stress-induced chaperones that facilitate the refolding and, thus, the degradation of abnormal proteins. Geranylgeranylacetone (GGA), a nontoxic antiulcer drug, has been shown to potently induce HSP expression in various tissues, including the central nervous system. In a cell model of SBMA, GGA increased the levels of Hsp70, Hsp90, and Hsp105 and inhibited cell death and the accumulation of pathogenic AR. Oral administration of GGA also up-regulated the expression of HSPs in the central nervous system of SBMA-transgenic mice and suppressed nuclear accumulation of the pathogenic AR protein, resulting in amelioration of polyglutamine-dependent neuromuscular phenotypes. These observations suggest that, although a high dose appears to be needed for clinical effects, oral GGA administration is a safe and promising therapeutic candidate for polyglutamine mediated neurodegenerative diseases, including SBMA. PMID- 16260739 TI - Innate control of adaptive immunity via remodeling of lymph node feed arteriole. AB - The adaptive immune system relies on rare cognate lymphocytes to detect pathogen derived antigens. Naive lymphocytes recirculate through secondary lymphoid organs in search of cognate antigen. Here, we show that the naive-lymphocyte recirculation pattern is controlled at the level of innate immune recognition, independent of antigen-specific stimulation. We demonstrate that inflammation induced lymphocyte recruitment to the lymph node is mediated by the remodeling of the primary feed arteriole, and that its physiological role is to increase the efficiency of screening for rare antigen-specific lymphocytes. Our data reveal a mechanism of innate control of adaptive immunity: by increasing the pool of naive lymphocytes for detection of foreign antigens via regulation of vascular input to the local lymph node. PMID- 16260740 TI - Site-specific recombinase and integrase activities of a conjugative relaxase in recipient cells. AB - Conjugative relaxases are the proteins that initiate bacterial conjugation by a site-specific cleavage of the transferred DNA strand. In vitro, they show strand transferase activity on single-stranded DNA, which suggests they may also be responsible for recircularization of the transferred DNA. In this work, we show that TrwC, the relaxase of plasmid R388, is fully functional in the recipient cell, as shown by complementation of an R388 trwC mutant in the recipient. TrwC transport to the recipient is also observed in the absence of DNA transfer, although it still requires the conjugative coupling protein. In addition to its role in conjugation, TrwC is able to catalyze site-specific recombination between two origin of transfer (oriT) copies. Mutations that abolish TrwC DNA strand transferase activity also abolish oriT-specific recombination. A plasmid containing two oriT copies resident in the recipient cell undergoes recombination when a TrwC-piloted DNA is conjugatively transferred into it. Finally, we show TrwC-dependent integration of the transferred DNA into a resident oriT copy in the recipient cell. Our results indicate that a conjugative relaxase is active once in the recipient cell, where it performs the nicking and strand-transfer reactions that would be required to recircularize the transferred DNA. This TrwC site-specific integration activity in recipient cells may lead to future biotechnological applications. PMID- 16260741 TI - Measles virus replication in lymphatic cells and organs of CD150 (SLAM) transgenic mice. AB - A transgenic mouse containing the complete human SLAM (hSLAM/CD150) gene, including its endogenous promoter for transcription, was generated by using human genomic DNA cloned into a bacterial artificial chromosome. hSLAM, the primary receptor for measles viruses (MV), was expressed on activated B, T, and dendritic cells with an expression profile equivalent to that of humans. We demonstrated that hSLAM(+) cells obtained from the transgenic mouse, including activated B, T, and dendritic cells, were susceptible to MV infection in a receptor-dependent manner. Evidence was provided for transient infection in the nasal lymph nodes of hSLAM(+) mice after intranasal inoculation. Virus was rapidly cleared without signs of secondary replication. To improve the efficiency of MV production, the hSLAM(+) mice were bred with mice having a Stat1-deficient background. These mice were more susceptible to MV infection and produced more virus particles. After intranasal and intraperitoneal inoculation of these mice with MV, infections of the thymus, spleen, nasal, mesenteric, and leg lymph nodes were detected. Upon necropsy, enlarged lymph nodes and spleen were apparent. Flow cytometric analysis showed that abnormally large numbers of mature neutrophils and natural killer cells caused the splenomegaly. The hSLAM transgenic mouse constitutes an improved rodent model for studying the interaction of MV with immune cells that more accurately reflects the infection pattern found in humans. PMID- 16260742 TI - Structural basis of the resistance of an insect carboxypeptidase to plant protease inhibitors. AB - Corn earworm (Helicoverpa zea), also called tomato fruitworm, is a common pest of many Solanaceous plants. This insect is known to adapt to the ingestion of plant serine protease inhibitors by using digestive proteases that are insensitive to inhibition. We have now identified a B-type carboxypeptidase of H. zea (CPBHz) insensitive to potato carboxypeptidase inhibitor (PCI) in corn earworm. To elucidate the structural features leading to the adaptation of the insect enzyme, the crystal structure of the recombinant CPBHz protein was determined by x-ray diffraction. CPBHz is a member of the A/B subfamily of metallocarboxypeptidases, which displays the characteristic metallocarboxypeptidase alpha/beta-hydrolase fold, and does not differ essentially from the previously described Helicoverpa armigera CPA, which is very sensitive to PCI. The data provide structural insight into several functional properties of CPBHz. The high selectivity shown by CPBHz for C-terminal lysine residues is due to residue changes in the S1' substrate specificity pocket that render it unable to accommodate the side chain of an arginine. The insensitivity of CPBHz to plant inhibitors is explained by the exceptional positioning of two of the main regions that stabilize other carboxypeptidase-PCI complexes, the beta8-alpha9 loop, and alpha7 together with the alpha7-alpha8 loop. The rearrangement of these two regions leads to a displacement of the active-site entrance that impairs the proper interaction with PCI. This report explains a crystal structure of an insect protease and its adaptation to defensive plant protease inhibitors. PMID- 16260743 TI - Brain lactate kinetics: Modeling evidence for neuronal lactate uptake upon activation. AB - A critical issue in brain energy metabolism is whether lactate produced within the brain by astrocytes is taken up and metabolized by neurons upon activation. Although there is ample evidence that neurons can efficiently use lactate as an energy substrate, at least in vitro, few experimental data exist to indicate that it is indeed the case in vivo. To address this question, we used a modeling approach to determine which mechanisms are necessary to explain typical brain lactate kinetics observed upon activation. On the basis of a previously validated model that takes into account the compartmentalization of energy metabolism, we developed a mathematical model of brain lactate kinetics, which was applied to published data describing the changes in extracellular lactate levels upon activation. Results show that the initial dip in the extracellular lactate concentration observed at the onset of stimulation can only be satisfactorily explained by a rapid uptake within an intraparenchymal cellular compartment. In contrast, neither blood flow increase, nor extracellular pH variation can be major causes of the lactate initial dip, whereas tissue lactate diffusion only tends to reduce its amplitude. The kinetic properties of monocarboxylate transporter isoforms strongly suggest that neurons represent the most likely compartment for activation-induced lactate uptake and that neuronal lactate utilization occurring early after activation onset is responsible for the initial dip in brain lactate levels observed in both animals and humans. PMID- 16260744 TI - Diversity and functional plasticity of eukaryotic selenoproteins: identification and characterization of the SelJ family. AB - Selenoproteins are a diverse group of proteins that contain selenocysteine (Sec), the 21st amino acid. In the genetic code, UGA serves as a termination signal and a Sec codon. This dual role has precluded the automatic annotation of selenoproteins. Recent advances in the computational identification of selenoprotein genes have provided a first glimpse of the size, functions, and phylogenetic diversity of eukaryotic selenoproteomes. Here, we describe the identification of a selenoprotein family named SelJ. In contrast to known selenoproteins, SelJ appears to be restricted to actinopterygian fishes and sea urchin, with Cys homologues only found in cnidarians. SelJ shows significant similarity to the jellyfish J1-crystallins and with them constitutes a distinct subfamily within the large family of ADP-ribosylation enzymes. Consistent with its potential role as a structural crystallin, SelJ has preferential and homogeneous expression in the eye lens in early stages of zebrafish development. A structural role for SelJ would be in contrast to the majority of known selenoenzymes. The unusually highly restricted phylogenetic distribution of SelJ, its specialization, and the comparative analysis of eukaryotic selenoproteomes reveal the diversity and functional plasticity of selenoproteins and point to a mosaic evolution of the use of Sec in proteins. PMID- 16260745 TI - High-efficiency transformation of Plasmodium falciparum by the lepidopteran transposable element piggyBac. AB - Functional analysis of the Plasmodium falciparum genome is restricted because of the limited ability to genetically manipulate this important human pathogen. We have developed an efficient transposon-mediated insertional mutagenesis method much needed for high-throughput functional genomics of malaria parasites. A drug selectable marker, human dihydrofolate reductase, added to the lepidopteran transposon piggyBac, transformed parasites by integration into the P. falciparum genome in the presence of a transposase-expressing helper plasmid. Multiple integrations occurred at the expected TTAA target sites throughout the genome of the parasite. We were able to transform P. falciparum with this piggyBac element at high frequencies, in the range of 10(-3), and obtain stable clones of insertional mutants in a few weeks instead of 6-12 months. Our results show that the piggyBac transposition system can be used as an efficient, random integration tool needed for large-scale, whole-genome mutagenesis of malaria parasites. The availability of such an adaptable genetic tool opens the way for much needed forward genetic approaches to study this lethal human parasite. PMID- 16260746 TI - Electrocatalytic hydrogen oxidation by an enzyme at high carbon monoxide or oxygen levels. AB - Use of hydrogen in fuel cells requires catalysts that are tolerant to oxygen and are able to function in the presence of poisons such as carbon monoxide. Hydrogen cycling catalysts are widespread in the bacterial world in the form of hydrogenases, enzymes with unusual active sites composed of iron, or nickel and iron, that are buried within the protein. We have established that the membrane bound hydrogenase from the beta-proteobacterium Ralstonia eutropha H16, when adsorbed at a graphite electrode, exhibits rapid electrocatalytic oxidation of hydrogen that is completely unaffected by carbon monoxide [at 0.9 bar (1 bar = 100 kPa), a 9-fold excess] and is inhibited only partially by oxygen. The practical significance of this discovery is illustrated with a simple fuel cell device, thus demonstrating the feasibility of future hydrogen-cycle technologies based on biological or biologically inspired electrocatalysts having high selectivity for hydrogen. PMID- 16260748 TI - Community inertia of Quaternary small mammal assemblages in North America. AB - One of the longest running debates in ecology is whether chance or determinism structures biotic communities, and this question is often studied by looking for the presence or absence of community inertia (lack of change) over time or space. Results have been equivocal. We adopted three tactics for a fresh approach: (i) allowing the answer to vary with the geographic, temporal, and taxonomic scale of study, (ii) using appropriate reference points for the amount of inertia in random biological systems, and (iii) using a robust approach for measurement of inertia. We examined fossil assemblages of mammalian communities across almost 1,000,000 years and at sites spanning approximately 3,500 km. We showed that in general there is good evidence for inertia but that the results change in a quantifiable fashion with taxonomic, spatial, and temporal scales. By using neutral theory we place a reference point on the degree of inertia and demonstrate that empirical mammalian communities show greater inertia than neutral communities over time scales >3,000 year. Although our results do not specifically reveal mechanism, they emphasize that deterministic forces are at work in structuring communities over millennia. PMID- 16260747 TI - Mechanisms regulating tissue-specific polarity of monocarboxylate transporters and their chaperone CD147 in kidney and retinal epithelia. AB - Proton-coupled monocarboxylate transporters (MCT) MCT1, MCT3, and MCT4 form heterodimeric complexes with the cell surface glycoprotein CD147 and exhibit tissue-specific polarized distributions that are essential for maintaining lactate and pH homeostasis. In the parenchymal epithelia of kidney, thyroid, and liver, MCT/CD147 heterocomplexes are localized in the basolateral membrane where they transport lactate out of or into the cell depending on metabolic conditions. A unique distribution of lactate transporters is found in the retinal pigment epithelium (RPE), which regulates lactate levels of the outer retina. In RPE, MCT1/CD147 is polarized to the apical membrane and MCT3/CD147 to the basolateral membrane. The mechanisms responsible for tissue-specific polarized distribution of MCTs are unknown. Here, we demonstrate that CD147 carries sorting information for polarized targeting of the MCT1/CD147 hetero-complexes in kidney and RPE cells. In contrast, MCT3 and MCT4 harbor dominant sorting information that cotargets CD147 to the basolateral membrane in both epithelia. RNA interference experiments show that MCT1 promotes CD147 maturation. Our results open a unique paradigm to study the molecular basis of tissue-specific polarity. PMID- 16260749 TI - Direct interaction of the human I-mfa domain-containing protein, HIC, with HIV-1 Tat results in cytoplasmic sequestration and control of Tat activity. AB - The primary function of the HIV-1 regulatory protein Tat, activation of transcription from the viral LTR, is highly regulated by complex interactions between Tat and a number of host cell proteins. Tat nuclear import, a process mediated by importin beta, is a prerequisite for its activity. Here, we report and characterize the interaction of the human inhibitor of MyoD family domain containing protein (I-mfa), HIC, with Tat at a biochemical and a functional level. This interaction was shown to occur in vivo and in vitro and to involve the nuclear localization signal and the transactivation responsive element binding domains of Tat and the I-mfa domain of HIC. Coexpression of HIC and Tat resulted in the down-regulation of transactivation of the HIV-1 LTR, and colocalization studies revealed the cytoplasmic sequestration of Tat by HIC. Functionally this sequestration appears to be the underlying mechanism of LTR transcriptional repression by HIC and represents a unique mechanism for the control of Tat activity and regulation of HIV-1 replication. PMID- 16260750 TI - Modeled regional climate change and California endemic oak ranges. AB - In the coming century, anthropogenic climate change will threaten the persistence of restricted endemic species, complicating conservation planning. Although most efforts to quantify potential shifts in species' ranges use global climate model (GCM) output, regional climate model (RCM) output may be better suited to predicting shifts by restricted species, particularly in regions with complex topography or other regionally important climate-forcing factors. Using a RCM based future climate scenario, we found that potential ranges of two California endemic oaks, Quercus douglasii and Quercus lobata, shrink considerably (to 59% and 54% of modern potential range sizes, respectively) and shift northward. This result is markedly different from that obtained by using a comparable GCM-based scenario, under which these species retain 81% and 73% of their modern potential range sizes, respectively. The difference between RCM- and GCM-based scenarios is due to greater warming and larger precipitation decreases during the growing season predicted by the RCM in these species' potential ranges. Based on the modeled regional climate change, <50% of protected land area currently containing these species is expected to contain them under a future midrange "business-as usual" path of greenhouse gas emissions. PMID- 16260751 TI - Long-range protein electron transfer observed at the single-molecule level: In situ mapping of redox-gated tunneling resonance. AB - A biomimetic long-range electron transfer (ET) system consisting of the blue copper protein azurin, a tunneling barrier bridge, and a gold single-crystal electrode was designed on the basis of molecular wiring self-assembly principles. This system is sufficiently stable and sensitive in a quasi-biological environment, suitable for detailed observations of long-range protein interfacial ET at the nanoscale and single-molecule levels. Because azurin is located at clearly identifiable fixed sites in well controlled orientation, the ET configuration parallels biological ET. The ET is nonadiabatic, and the rate constants display tunneling features with distance-decay factors of 0.83 and 0.91 A(-1) in H(2)O and D(2)O, respectively. Redox-gated tunneling resonance is observed in situ at the single-molecule level by using electrochemical scanning tunneling microscopy, exhibiting an asymmetric dependence on the redox potential. Maximum resonance appears around the equilibrium redox potential of azurin with an on/off current ratio of approximately 9. Simulation analyses, based on a two step interfacial ET model for the scanning tunneling microscopy redox process, were performed and provide quantitative information for rational understanding of the ET mechanism. PMID- 16260752 TI - Semantic congruity affects numerical judgments similarly in monkeys and humans. AB - Monkeys (Macaca mulatta) were trained to order visual arrays based on their number of elements and to conditionally choose the array with the larger or smaller number of elements dependent on a color cue. When the screen background was red, monkeys were reinforced for choosing the smaller numerical value first. When the screen background was blue, monkeys were reinforced for choosing the larger numerical value first. Monkeys showed a semantic congruity effect analogous to that reported for human comparison judgments. Specifically, decision time was systematically influenced by the semantic congruity between the cue ("choose smaller" or "choose larger") and the magnitude of the choice stimuli (small or large numbers of dots). This finding demonstrates a semantic congruity effect in a nonlinguistic animal and provides strong evidence for an evolutionarily primitive magnitude-comparison algorithm common to humans and monkeys. PMID- 16260753 TI - Three genomes differentially contribute to the biosynthesis of benzoxazinones in hexaploid wheat. AB - Hexaploid wheat (Triticum aestivum) accumulates benzoxazinones (Bxs) as defensive compounds. Previously, we found that five Bx biosynthetic genes, TaBx1-TaBx5, are located on each of the three genomes (A, B, and D) of hexaploid wheat. In this study, we isolated three homoeologous cDNAs of each TaBx gene to estimate the contribution of individual homoeologous TaBx genes to the biosynthesis of Bxs in hexaploid wheat. We analyzed their transcript levels by homoeolog- or genome specific quantitative RT-PCR and the catalytic properties of their translation products by kinetic analyses using recombinant TaBX enzymes. The three homoeologs were transcribed differentially, and the ratio of the individual homoeologous transcripts to total homoeologous transcripts also varied with the tissue, i.e., shoots or roots, as well as with the developmental stage. Moreover, the translation products of the three homoeologs had different catalytic properties. Some TaBx homoeologs were efficiently transcribed, but the translation products showed only weak enzymatic activities, which inferred their weak contribution to Bx biosynthesis. Considering the transcript levels and the catalytic properties collectively, we concluded that the homoeologs on the B genome generally contributed the most to the Bx biosynthesis in hexaploid wheat, especially in shoots. In tetraploid wheat and the three diploid progenitors of hexaploid wheat, the respective transcript levels of the TaBx homoeologs were similar in ratio to those observed in hexaploid wheat. This result indicates that the genomic bias in the transcription of the TaBx genes in hexaploid wheat originated in the diploid progenitors and has been retained through the polyploidization. PMID- 16260755 TI - A novel approach to structural alignment using realistic structural and environmental information. AB - In the era of structural genomics, it is necessary to generate accurate structural alignments in order to build good templates for homology modeling. Although a great number of structural alignment algorithms have been developed, most of them ignore intermolecular interactions during the alignment procedure. Therefore, structures in different oligomeric states are barely distinguishable, and it is very challenging to find correct alignment in coil regions. Here we present a novel approach to structural alignment using a clique finding algorithm and environmental information (SAUCE). In this approach, we build the alignment based on not only structural coordinate information but also realistic environmental information extracted from biological unit files provided by the Protein Data Bank (PDB). At first, we eliminate all environmentally unfavorable pairings of residues. Then we identify alignments in core regions via a maximal clique finding algorithm. Two extreme value distribution (EVD) form statistics have been developed to evaluate core region alignments. With an optional extension step, global alignment can be derived based on environment-based dynamic programming linking. We show that our method is able to differentiate three-dimensional structures in different oligomeric states, and is able to find flexible alignments between multidomain structures without predetermined hinge regions. The overall performance is also evaluated on a large scale by comparisons to current structural classification databases as well as to other alignment methods. PMID- 16260754 TI - Selection of ribozymes that catalyse multiple-turnover Diels-Alder cycloadditions by using in vitro compartmentalization. AB - In vitro compartmentalization (IVC) has previously been used to evolve protein enzymes. Here, we demonstrate how IVC can be applied to select RNA enzymes (ribozymes) for a property that has previously been unselectable: true intermolecular catalysis. Libraries containing 10(11) ribozyme genes are compartmentalized in the aqueous droplets of a water-in-oil emulsion, such that most droplets contain no more than one gene, and transcribed in situ. By coencapsulating the gene, RNA, and the substrates/products of the catalyzed reaction, ribozymes can be selected for all enzymatic properties: substrate recognition, product formation, rate acceleration, and turnover. Here we exploit the complementarity of IVC with systematic evolution of ligands by exponential enrichment (SELEX), which allows selection of larger libraries (>/=10(15)) and for very small rate accelerations (k(cat)/k(uncat)) but only selects for intramolecular single-turnover reactions. We selected approximately 10(14) random RNAs for Diels-Alderase activity with five rounds of SELEX, then six to nine rounds with IVC. All selected ribozymes catalyzed the Diels-Alder reaction in a truly bimolecular fashion and with multiple turnover. Nearly all ribozymes selected by using eleven rounds of SELEX alone contain a common catalytic motif. Selecting with SELEX then IVC gave ribozymes with significant sequence variations in this catalytic motif and ribozymes with completely novel motifs. Interestingly, the catalytic properties of all of the selected ribozymes were quite similar. The ribozymes are strongly product inhibited, consistent with the Diels-Alder transition state closely resembling the product. More efficient Diels Alderases may need to catalyze a second reaction that transforms the product and prevents product inhibition. PMID- 16260756 TI - Pneumococcal phosphorylcholine esterase, Pce, contains a metal binuclear center that is essential for substrate binding and catalysis. AB - The phosphorylcholine esterase from Streptococcus pneumoniae, Pce, catalyzes the hydrolysis of phosphorylcholine residues from teichoic and lipoteichoic acids attached to the bacterial envelope and comprises a globular N-terminal catalytic module containing a zinc binuclear center and an elongated C-terminal choline binding module. The dependence of Pce activity on the metal/enzyme stoichiometry shows that the two equivalents of zinc are essential for the catalysis, and stabilize the catalytic module through a complex metal-ligand coordination network. The pH dependence of Pce activity toward the alternative substrate p nitrophenylphosphorylcholine (NPPC) shows that k(cat) and k(cat)/K(m) depend on the protonation state of two protein residues that can be tentatively assigned to the ionization of the metal-bound water (hydrogen bonded to D89) and to H228. Maximum activity requires deprotonation of both groups, although the catalytic efficiency is optimum for the single deprotonated form. The drastic reduction of activity in the H90A mutant, which still binds two Zn2+ ions at neutral pH, indicates that Pce activity also depends on the geometry of the metallic cluster. The denaturation heat capacity profile of Pce exhibits two peaks with T(m) values of 39.6 degrees C (choline-binding module) and 60.8 degrees C (catalytic module). The H90A mutation reduces the high-temperature peak by about 10 degrees C. Pce is inhibited in the presence of 1 mM zinc, but this inhibition depends on pH, buffer, and substrate species. A reaction mechanism is proposed on the basis of kinetic data, the structural model of the Pce:NPPC complex, and the currently accepted mechanism for other Zn-metallophosphoesterases. PMID- 16260757 TI - Disruption of an intermonomer salt bridge in the p53 tetramerization domain results in an increased propensity to form amyloid fibrils. AB - We describe in molecular detail how disruption of an intermonomer salt bridge (Arg337-Asp352) leads to partial destabilization of the p53 tetramerization domain and a dramatically increased propensity to form amyloid fibrils. At pH 4.0 and 37 degrees C, a p53 tetramerization domain mutant (p53tet-R337H), associated with adrenocortical carcinoma in children, readily formed amyloid fibrils, while the wild-type (p53tet-wt) did not. We characterized these proteins by equilibrium denaturation, 13C(alpha) secondary chemical shifts, (1H)-15N heteronuclear NOEs, and H/D exchange. Although p53tet-R337H was thermodynamically less stable, NMR data indicated that the two proteins had similar secondary structure and molecular dynamics. NMR derived pK(a) values indicated that at low pH the R337H mutation partially disrupted an intermonomer salt bridge. Backbone H/D exchange results showed that for at least a small population of p53tet-R337H molecules disruption of this salt bridge resulted in partial destabilization of the protein. It is proposed that this decrease in p53tet-R337H stability resulted in an increased propensity to form amyloid fibrils. PMID- 16260759 TI - Evolution of an acylase active on cephalosporin C. AB - Semisynthetic cephalosporins are synthesized from 7-amino cephalosporanic acid, which is produced by chemical deacylation or by a two-step enzymatic process of the natural antibiotic cephalosporin C. The known acylases take glutaryl-7-amino cephalosporanic acid as a primary substrate, and their specificity and activity are too low for cephalosporin C. Starting from a known glutaryl-7-amino cephalosporanic acid acylase as the protein scaffold, an acylase gene optimized for expression in Escherichia coli and for molecular biology manipulations was designed. Subsequently we used error-prone PCR mutagenesis, a molecular modeling approach combined with site-saturation mutagenesis, and site-directed mutagenesis to produce enzymes with a cephalosporin C/glutaryl-7-amino cephalosporanic acid catalytic efficiency that was increased up to 100-fold, and with a significant and higher maximal activity on cephalosporin C as compared to glutaryl-7-amino cephalosporanic acid (e.g., 3.8 vs. 2.7 U/mg protein, respectively, for the A215Y H296S-H309S mutant). Our data in a bioreactor indicate an ~90% conversion of cephalosporin C to 7-amino-cephalosporanic acid in a single deacylation step. The evolved acylase variants we produced are enzymes with a new substrate specificity, not found in nature, and represent a hallmark for industrial production of 7-amino cephalosporanic acid. PMID- 16260758 TI - Solution structure of (gamma)S-crystallin by molecular fragment replacement NMR. AB - The solution structure of murine gammaS-crystallin (gammaS) has been determined by multidimensional triple resonance NMR spectroscopy, using restraints derived from two sets of dipolar couplings, recorded in different alignment media, and supplemented by a small number of NOE distance restraints. gammaS consists of two topologically similar domains, arranged with an approximate twofold symmetry, and each domain shows close structural homology to closely related (approximately 50% sequence identity) domains found in other members of the gamma-crystallin family. Each domain consists of two four-strand "Greek key" beta-sheets. Although the domains are tightly anchored to one another by the hydrophobic surfaces of the two inner Greek key motifs, the N-arm, the interdomain linker and several turn regions show unexpected flexibility and disorder in solution. This may contribute entropic stabilization to the protein in solution, but may also indicate nucleation sites for unfolding or other structural transitions. The method used for solving the gammaS structure relies on the recently introduced molecular fragment replacement method, which capitalizes on the large database of protein structures previously solved by X-ray crystallography and NMR. PMID- 16260761 TI - Functional expression of eukaryotic membrane proteins in Lactococcus lactis. AB - The overproduction of eukaryotic membrane proteins is a major impediment in their structural and functional characterization. Here we have used the nisin-inducible expression system of Lactococcus lactis for the overproduction of 11 mitochondrial transport proteins from yeast. They were expressed at high levels in a functional state in the cytoplasmic membrane. The results also show that the level of expression is influenced by the N-terminal regions of the transporters. Expression levels were improved >10-fold either by replacing or truncating these regions or by adding lactococcal signal peptides. The observed expression levels are now compatible with a realistic exploration of crystallization conditions. The lactococcal expression system may be used for the high-throughput functional characterization of eukaryotic membrane proteins and structural genomics. PMID- 16260760 TI - Distinct interaction modes of an AKAP bound to two regulatory subunit isoforms of protein kinase A revealed by amide hydrogen/deuterium exchange. AB - The structure of an AKAP docked to the dimerization/docking (D/D) domain of the type II (RIIalpha) isoform of protein kinase A (PKA) has been well characterized, but there currently is no detailed structural information of an AKAP docked to the type I (RIalpha) isoform. Dual-specific AKAP2 (D-AKAP2) binds in the nanomolar range to both isoforms and provided us with an opportunity to characterize the isoform-selective nature of AKAP binding using a common docked ligand. Hydrogen/deuterium (H/D) exchange combined with mass spectrometry (DXMS) was used to probe backbone structural changes of an alpha-helical A-kinase binding (AKB) motif from D-AKAP2 docked to both RIalpha and RIIalpha D/D domains. The region of protection upon complex formation and the magnitude of protection from H/D exchange were determined for both interacting partners in each complex. The backbone of the AKB ligand was more protected when bound to RIalpha compared to RIIalpha, suggesting an increased helical stabilization of the docked AKB ligand. This combined with a broader region of backbone protection induced by the AKAP on the docking surface of RIalpha indicated that there were more binding constraints for the AKB ligand when bound to RIalpha. This was in contrast to RIIalpha, which has a preformed, localized binding surface. These distinct modes of AKAP binding may contribute to the more discriminating nature of the RIalpha AKAP-docking surface. DXMS provides valuable structural information for understanding binding specificity in the absence of a high-resolution structure, and can readily be applied to other protein-ligand and protein-protein interactions. PMID- 16260762 TI - Modulation of oxygen binding to insect hemoglobins: the structure of hemoglobin from the botfly Gasterophilus intestinalis. AB - Hemoglobins (Hbs) reversibly bind gaseous diatomic ligands (e.g., O2) as the sixth heme axial ligand of the penta-coordinate deoxygenated form. Selected members of the Hb superfamily, however, display a functionally relevant hexa coordinate heme Fe atom in their deoxygenated state. Endogenous heme hexa coordination is generally provided in these Hbs by the E7 residue (often His), which thus modulates accessibility to the heme distal pocket and reactivity of the heme toward exogenous ligands. Such a pivotal role of the E7 residue is prominently shown by analysis of the functional and structural properties of insect Hbs. Here, we report the 2.6 A crystal structure of oxygenated Gasterophilus intestinalis Hb1, a Hb known to display a penta-coordinate heme in the deoxygenated form. The structure is analyzed in comparison with those of Drosophila melanogaster Hb, exhibiting a hexa-coordinate heme in its deoxygenated derivative, and of Chironomus thummi thummi HbIII, which displays a penta coordinate heme in the deoxygenated form. Despite evident structural differences in the heme distal pockets, the distinct molecular mechanisms regulating O2 binding to the three insect Hbs result in similar O(2 affinities (P50 values ranging between 0.12 torr and 0.46 torr). PMID- 16260763 TI - Crystal structures of T cell receptor (beta) chains related to rheumatoid arthritis. AB - The crystal structures of the Vbeta17+ beta chains of two human T cell receptors (TCRs), originally derived from the synovial fluid (SF4) and tissue (C5-1) of a patient with rheumatoid arthritis (RA), have been determined in native (SF4) and mutant (C5-1(F104-->Y/C187-->S)) forms, respectively. These TCR beta chains form homo-dimers in solution and in crystals. Structural comparison reveals that the main-chain conformations in the CDR regions of the C5-1 and SF4 Vbeta17 closely resemble those of a Vbeta17 JM22 in a bound form; however, the CDR3 region shows different conformations among these three Vbeta17 structures. At the side-chain level, conformational differences were observed at the CDR2 regions between our two ligand-free forms and the bound JM22 form. Other significant differences were observed at the Vbeta regions 8-12, 40-44, and 82-88 between C5-1/SF4 and JM22 Vbeta17, implying that there is considerable variability in the structures of very similar beta chains. Structural alignments also reveal a considerable variation in the Vbeta-Cbeta associations, and this may affect ligand recognition. The crystal structures also provide insights into the structure basis of T cell recognition of Mycoplasma arthritidis mitogen (MAM), a superantigen that may be implicated in the development of human RA. Structural comparisons of the Vbeta domains of known TCR structures indicate that there are significant similarities among Vbeta regions that are MAM-reactive, whereas there appear to be significant structural differences among those Vbeta regions that lack MAM-reactivity. It further reveals that CDR2 and framework region (FR) 3 are likely to account for the binding of TCR to MAM. PMID- 16260764 TI - High yield bacterial expression of active c-Abl and c-Src tyrosine kinases. AB - The Abl and Src tyrosine kinases are key signaling proteins that are of considerable interest as drug targets in cancer and many other diseases. The regulatory mechanisms that control the activity of these proteins are complex, and involve large-scale conformational changes in response to phosphorylation and other modulatory signals. The success of the Abl inhibitor imatinib in the treatment of chronic myelogenous leukemia has shown the potential of kinase inhibitors, but the rise of drug resistance in patients has also shown that drugs with alternative modes of binding to the kinase are needed. The detailed understanding of mechanisms of protein-drug interaction and drug resistance through biophysical methods demands a method for the production of active protein on the milligram scale. We have developed a bacterial expression system for the kinase domains of c-Abl and c-Src, which allows for the quick expression and purification of active wild-type and mutant kinase domains by coexpression with the YopH tyrosine phosphatase. This method makes practical the use of isotopic labeling of c-Abl and c-Src for NMR studies, and is also applicable for constructs containing the SH2 and SH3 domains of the kinases. PMID- 16260765 TI - Solution structure of the Escherichia coli protein ydhR: a putative mono oxygenase. AB - YdhR is a 101-residue conserved protein from Escherichia coli. Sequence searches reveal that the protein has >50% identity to proteins found in a variety of other bacterial genomes. Using size exclusion chromatography and fluorescence spectroscopy, we determined that ydhR exists in a dimeric state with a dissociation constant of approximately 40 nM. The three-dimensional structure of dimeric ydhR was determined using NMR spectroscopy. A total of 3400 unambiguous NOEs, both manually and automatically assigned, were used for the structure calculation that was refined using an explicit hydration shell. A family of 20 structures was obtained with a backbone RMSD of 0.48 A for elements of secondary structure. The structure reveals a dimeric alpha,beta fold characteristic of the alpha+beta barrel superfamily of proteins. Bioinformatic approaches were used to show that ydhR likely belongs to a recently identified group of mono-oxygenase proteins that includes ActVA-Orf6 and YgiN and are involved in the oxygenation of polyaromatic ring compounds. PMID- 16260766 TI - The crystal structure of a novel SAM-dependent methyltransferase PH1915 from Pyrococcus horikoshii. AB - The S-adenosyl-L-methionine (SAM)-dependent methyltransferases represent a diverse and biologically important class of enzymes. These enzymes utilize the ubiquitous methyl donor SAM as a cofactor to methylate proteins, small molecules, lipids, and nucleic acids. Here we present the crystal structure of PH1915 from Pyrococcus horikoshii OT3, a predicted SAM-dependent methyltransferase. This protein belongs to the Cluster of Orthologous Group 1092, and the presented crystal structure is the first representative structure of this protein family. Based on sequence and 3D structure analysis, we have made valuable functional insights that will facilitate further studies for characterizing this group of proteins. Specifically, we propose that PH1915 and its orthologs are rRNA- or tRNA-specific methyltransferases. PMID- 16260767 TI - Automated identification and quantification of protein phosphorylation sites by LC/MS on a hybrid triple quadrupole linear ion trap mass spectrometer. AB - Complete phosphorylation mapping of protein kinases was successfully undertaken using an automated LC/MS/MS approach. This method uses the direct combination of triple quadrupole and ion trapping capabilities in a hybrid triple quadrupole linear ion trap to selectively identify and sequence phosphorylated peptides. In particular, the use of a precursor ion scan of m/z -79 in negative ion mode followed by an ion trap high resolution scan (an enhanced resolution scan) and a high sensitivity MS/MS scan (enhanced product ion scan) in positive mode is a very effective method for identifying phosphorylation sites in proteins at low femtomole levels. Coupling of this methodology with a stable isotope N-terminal labeling strategy using iTRAQtrade mark reagents enabled phosphorylation mapping and relative protein phosphorylation levels to be determined between the active and inactive forms of the protein kinase MAPKAPK-1 in the same LC/MS run. PMID- 16260768 TI - Egg predation fuels unique species association at deep-sea hydrocarbon seeps. PMID- 16260769 TI - Endosymbiotic yeast maternally transmitted in a marine sponge. AB - The detection of an endosymbiotic yeast in demosponges of the genus Chondrilla described here records the first such association within the phylum Porifera. The symbiont, interpreted as a yolk body in previous ultrastructural studies, is a chitinous-walled fission yeast. Chitin was detected by an immunocytochemical technique that labels its beta-1,4-N-acetyl-D-glucosamine residues. Abundant symbiotic yeast cells (4.4 +/- 2.3 cells per 10 microm2) transmitted from the soma through the oocytes to the fertilized eggs are directly propagated by vertical transmission in the female. Vertically transmitted yeast were detected in three Chondrilla species with disjunct biogeographical distributions: the Mediterranean, the Caribbean, and the Australian Pacific. Apparently these yeasts are not present in other demosponge genera. Therefore, the fungal endosymbiosis most likely evolved before or during the diversification of the genus Chondrilla. PMID- 16260770 TI - Self- and cross-fertilization in the solitary ascidian Ciona savignyi. AB - Solitary ascidians are hermaphrodites that release sperm and eggs simultaneously. However, many species are self-sterile, owing to a self/non-self recognition system operating at the outer surface of the chorion during sperm-egg interaction. In Ciona intestinalis, self-incompatibility is thought to have a genetic basis. Here, we report a survey of the self-fertilization potential of a Santa Barbara, California, population of Ciona savignyi, a close relative of C. intestinalis. We found that, in contrast to reports on C. intestinalis, C. savignyi is highly self-fertile. However, using two nonlethal recessive mutant strains, aimless (aim) and immaculate (imc), and a stable transgenic strain that expresses green fluorescent protein (GFP) in the notochord to follow offspring genotype, we demonstrate that non-self sperm outcompete self-sperm in fertilization competition assays. When the chorion was removed, both self- and non-self sperm performed equally well in the competition assay. Thus the non self/self gamete recognition in C. savignyi is not absolute but relative, and is mediated by one or more components in the chorion. We discuss the significance of this finding in the context of natural populations in the wild, where individuals of C. savignyi are typically found growing in large groups that spawn in unison and where self-fertilization would be expected to be very rare. PMID- 16260771 TI - Gamete compatibility and sperm competition affect paternity and hybridization between sympatric Asterias sea stars. AB - Gamete interactions may strongly influence speciation and hybridization in sympatric broadcast-spawning marine invertebrates. We examined the role of gamete compatibility in species integrity using cross-fertilization studies between sympatric Asterias sea stars from a secondary contact zone in the northwest Atlantic. In crosses between single males and single females, gametes of both species were compatible and produced viable, fertile hybrid offspring, but with considerable variation in the receptivity of eggs to heterospecific sperm. Differential compatibility of heterospecific gametes was detected in sperm competition studies in which we used a nuclear DNA marker to assign paternity to larval offspring. Several families showed conspecific sperm precedence in A. forbesi eggs, and one family showed competitive superiority of A. forbesi sperm fertilizing A. rubens eggs. Gametic interactions are an important component of prezygotic reproductive isolation in sympatric Asterias. The interaction between gametes of these closely related sea stars is consistent with the function of gamete recognition systems that are known to mediate fertilization success and speciation in other marine invertebrates. PMID- 16260773 TI - Infaunal hydraulics generate porewater pressure signals. AB - Many activities by infauna, including burrowing and feeding, involve hydraulic mechanisms. We expected these activities to generate low-frequency pressure waves that would propagate through sediments and be detectable at some distance from the source. Pressure sensors in intertidal sediments recorded large-amplitude porewater pressure signals. Laboratory recordings of single individuals allowed us to identify characteristic signals of arenicolid and nereidid polychaetes and tellinid bivalves. In the bivalve Macoma nasuta, these high-amplitude signals were associated with burrowing, expulsion of pseudofeces, and siphon relocation. In the polychaetes Neanthes brandti and Abarenicola pacifica, the high-amplitude pressure signals were associated with burrowing, burrow construction, burrow ventilation, and defecation. These signals were detectable in the field at distances of at least 20 cm. Since the waveforms are species-specific as well as activity-specific, they may provide a mechanism for prey detection, for predator avoidance, for competitor detection, and perhaps even for mate detection. PMID- 16260772 TI - Developmental plasticity in Macrophiothrix brittlestars: are morphologically convergent larvae also convergently plastic? AB - The pluteus larval forms of sea urchins (echinoids) and brittlestars (ophiuroids) use an internal skeleton to project arms that bear a long ciliated band used in swimming and feeding. The length of this ciliated band influences rates of maximum food clearance for larvae of both echinoderm classes and affects rates of growth and development in the plankton. Phylogenetic and morphological evidence, however, tend to support the view that the pluteus morphologies of the two classes are independently derived. Studies with echinoplutei have shown that investment in skeletal growth and ciliated band length changes in response to food conditions, with poorly fed larvae investing more in growth of the larval skeleton and arms either absolutely or in relation to other larval or developing postlarval structures. We present evidence for similar plasticity of skeletal growth in ophioplutei. We examined four species in the brittlestar genus Macrophiothrix that spanned a 3.8-fold range in egg size. Sibling larvae in 14 male-female crosses were reared with high (H) or low (L) food rations, and measurements were recorded for five skeletal arm rods and three non-arm body dimensions. The expression of adaptive plasticity (significantly longer arms in L versus H cultures on a given day) was apparent for most crosses in M. koehleri, the species with the smallest egg size. In the single cross for M. longipeda, larvae from L cultures had longer arms for their body length or stomach width than did larvae from H cultures. In these cases, plasticity was similar in timing, persistence, and magnitude to previously published results from echinoplutei. If internal skeletons are independently derived in the two classes, then plasticity in the expression of this homoplastic trait may itself be homoplastic. PMID- 16260774 TI - Piscivorous behavior of a temperate cone snail, Conus californicus. AB - Most of the more than 500 species of predatory marine snails in the genus Conus are tropical or semitropical, and nearly all are thought to be highly selective regarding type of prey. Conus californicus Hinds, 1844, is unusual in that it is endemic to the North American Pacific coast and preys on a large variety of benthic organisms, primarily worms and other molluscs, and also scavenges. We studied the feeding behavior of C. californicus in captivity and found that it regularly killed and consumed live prickleback fishes (Cebidichthys violaceus and Xiphister spp.). Predation involved two behavioral methods similar to those employed by strictly piscivorous relatives. One method utilized stings delivered by radular teeth; the other involved engulfing the prey without stinging. Both methods were commonly used in combination, and individual snails sometimes employed multiple stings to subdue a fish. During the course of the study, snails became aroused by the presence of live fish more quickly, as evidenced by more rapid initiation of hunting behavior. Despite this apparent adaptation, details of prey-capture techniques and effectiveness of stings remained similar over the same period. PMID- 16260775 TI - Muscle organization of the cubozoan jellyfish Tripedalia cystophora Conant 1897. AB - The musculature of the cubomedusa Tripedalia cystophora was investigated using immunohistochemical staining with an anti-actin antibody and histochemical staining with fluorescent phalloidin. The subumbrella is lined with a sheet of circular, striated muscle that is interrupted at the perradii, and by the nerve ring. The sheet is continuous with circular, striated muscle of the velarium, which turns radially on each face of the four velarial frenula. Perradial strips of smooth muscle run radially from just above the level of the rhopalia into the manubrium and lips. The strips give off perpendicular offshoots that run a short distance in parallel with the circular swim muscle. Musculature of the tentacles and pedalia is longitudinal and limited to the oral side of the pedalia. The pedalial muscle connects with bundles of smooth muscle that runs circularly from the tentacle base well into the subumbrella. The arrangement of striated muscle in the frenula suggests that these structures may function in directional nozzle formation of the velarium during turning. In addition, the perpendicular branching of the radial strips and the circular extensions of pedalial muscle may function in hinge formation to aid bending of the pedalia and tentacles into the subumbrella during feeding and protective responses. PMID- 16260776 TI - Egr-1 induces the expression of its corepressor nab2 by activation of the nab2 promoter thereby establishing a negative feedback loop. AB - The transcription factor Egr-1 regulates the expression of numerous genes involved in differentiation, growth, and in response to environmental signals. Egr-1 activity is modulated in part through the binding of corepressors Nab1 and Nab2. Nab2 appears crucial for controlling Egr-1-mediated transactivation because it is a delayed early response gene, induced by the same stimuli that induce the immediate early gene Egr-1. To identify important elements regulating Nab2 expression, we cloned the human Nab2 gene and investigated the 5'-region. The TATA- and initiator-less Nab2 promoter, located from -679 to -74 bp, contains a total of 11 Egr binding sites, including a cluster of multiple overlapping Egr/Sp1 sites between -329 and -260 bp. This region is critical for basal promoter activity as well as for maximum induction by phorbol esters. Electromobility shifts show that Sp1 binds to this region in normal and stimulated cells, whereas stimulation induces binding of Egr-1. In addition Egr-1 activates the Nab2 promoter in a pattern similar to phorbol esters, suggesting that Egr-1 is a major inducer of protein kinase C-mediated Nab2 induction. Depletion of Egr-1 by each of two distinct Egr-1 short-interfering RNAs reduces Nab2 expression and inducibility, confirming that Egr-1 is an important regulator of Nab2 expression. Transfection experiments show that Egr-1-induced Nab2 promoter activity is itself repressed by Nab2. These results indicate that Egr-1 mediates the induction of its own repressor, thereby preventing a permanent transactivation of Egr-1 target genes and a damaging overreaction in response to environmental signals. PMID- 16260777 TI - Decreased subunit stability as a novel mechanism for potassium current impairment by a KCNQ2 C terminus mutation causing benign familial neonatal convulsions. AB - KCNQ2 and KCNQ3 K+ channel subunits underlie the muscarinic-regulated K+ current (I(KM)), a widespread regulator of neuronal excitability. Mutations in KCNQ2- or KCNQ3-encoding genes cause benign familiar neonatal convulsions (BFNCs), a rare autosomal-dominant idiopathic epilepsy of the newborn. In the present study, we have investigated, by means of electrophysiological, biochemical, and immunocytochemical techniques in transiently transfected cells, the consequences prompted by a BFNC-causing 1-bp deletion (2043deltaT) in the KCNQ2 gene; this frameshift mutation caused the substitution of the last 163 amino acids of the KCNQ2 C terminus and the extension of the subunit by additional 56 residues. The 2043deltaT mutation abolished voltage-gated K+ currents produced upon homomeric expression of KCNQ2 subunits, dramatically reduced the steady-state cellular levels of KCNQ2 subunits, and prevented their delivery to the plasma membrane. Metabolic labeling experiments revealed that mutant KCNQ2 subunits underwent faster degradation; 10-h treatment with the proteasomal inhibitor MG132 (20 microm) at least partially reversed such enhanced degradation. Co-expression with KCNQ3 subunits reduced the degradation rate of mutant KCNQ2 subunits and led to their expression on the plasma membrane. Finally, co-expression of KCNQ2 2043deltaT together with KCNQ3 subunits generated functional voltage-gated K+ currents having pharmacological and biophysical properties of heteromeric channels. Collectively, the present results suggest that mutation-induced reduced stability of KCNQ2 subunits may cause epilepsy in neonates. PMID- 16260778 TI - An hGCN5/TRRAP histone acetyltransferase complex co-activates BRCA1 transactivation function through histone modification. AB - It is well established that genetic mutations that impair BRCA1 function predispose women to early onset of breast and ovarian cancer. However, the co regulatory factors that support normal BRCA1 functions remain to be identified. Using a biochemical approach to search for such co-regulatory factors, we identified hGCN5, TRRAP, and hMSH2/6 as BRCA1-interacting proteins. Genetic mutations in the C-terminal transactivation domain of BRCA1, as found in breast cancer patients (Chapman, M. S., and Verma, I. M. (1996) Nature 382, 678-679), caused the loss of physical interaction between BRCA1 and TRRAP and significantly reduced the co-activation of BRCA1 transactivation function by hGCN5/TRRAP. The reported transcriptional squelching between BRCA1 and estrogen receptor alpha (Fan, S., Wang, J., Yuan, R., Ma, Y., Meng, Q., Erdos, M. R., Pestell, R. G., Yuan, F., Auborn, K. J., Goldberg, I. D., and Rosen, E. M. (1999) Science 284, 1354-1356) was rescued by the overexpression of TRRAP or hGCN5. Histone acetyltransferase hGCN5 activity appeared to be indispensable for coregulator complex function in both BRCA1-mediated gene regulation and DNA repair. Biochemical purification of the hGCN5/TRRAP-containing complex suggested that hGCN5/TRRAP formed a complex with hMSH2/hMSH6, presumably as a novel subclass of hGCN5/TRRAP-containing known TFTC (TBP-free TAF-containing)-type histone acetyltransferase complex (hTFTC, hPCAF, and hSTAGA) (Yanagisawa, J., Kitagawa, H., Yanagida, M., Wada, O., Ogawa, S., Nakagomi, M., Oishi, H., Yamamoto, Y., Nagasawa, H., McMahon, S. B., Cole, M. D., Tora, L., Takahashi, N., and Kato, S. (2002) Mol. Cell 9, 553-562). Unlike other subclasses, the isolated complex harbored a previously unknown combination of components including hMSH2 and hMSH6, major components of the BRCA1 genome surveillance repair complex (BASC). Thus, our results suggested that the multiple BRCA1 functions require a novel hGCN5/TRRAP histone acetyltransferase complex subclass. PMID- 16260779 TI - The streptococcal lipoprotein rotamase A (SlrA) is a functional peptidyl-prolyl isomerase involved in pneumococcal colonization. AB - Streptococcus pneumoniae expresses two surface-exposed lipoproteins, PpmA and SlrA, which share homology with distinct families of peptidyl-prolyl isomerases (PPIases). In this study, we demonstrated for the first time that the lipoprotein cyclophilin, SlrA, can catalyze the cis-trans isomerization of proline containing tetrapeptides and that SlrA contributes to pneumococcal colonization. The substrate specificity of SlrA is typical for prokaryotic and eukaryotic cyclophilins, with Suc-Ala-Ala-Pro-Phe-p-nitroanilide (pNA) being the most rapidly catalyzed substrate. In a mouse pneumonia model the slrA knock-out D39DeltaslrA did not cause significant differences in the survival times of mice compared with the isogenic wild-type strain. In contrast, a detailed analysis of bacterial outgrowth over time in the nasopharynx, airways, lungs, blood, and spleen showed a rapid elimination of slrA mutants from the upper airways but did not reveal significant differences in the lungs, blood, and spleen. These results suggested that SlrA is involved in colonization but does not contribute significantly to invasive pneumococcal disease. In cell culture infection experiments, the absence of SlrA impaired adherence to pneumococcal disease specific epithelial and endothelial non-professional cell lines. Adherence of the slrA mutant could not be restored by exogenously added SlrA. Strikingly, deficiency in SlrA did not reduce binding activity to host target proteins, but resulted in enhanced uptake by professional phagocytes. In conclusion, SlrA is a functional, cyclophilin-type PPIase and contributes to pneumococcal virulence in the first stage of infection, namely, colonization of the upper airways, most likely by modulating the biological function of important virulence proteins. PMID- 16260781 TI - Receptor activator of NF-kappaB (RANK) cytoplasmic motif, 369PFQEP373, plays a predominant role in osteoclast survival in part by activating Akt/PKB and its downstream effector AFX/FOXO4. AB - Receptor activator of NF-kappaB ligand (RANKL) plays a crucial role in osteoclast differentiation, function, and survival. RANKL exerts its effect by activating its receptor RANK (receptor activator of NF-kappaB), which recruits various intracellular signaling molecules via specific motifs in its cytoplasmic tail. Previously, we identified three RANK cytoplasmic motifs (Motif 1, 369PFQEP373; Motif 2, 559PVQEET564; and Motif 3, 604PVQEQG609) mediating osteoclast formation and function. Here, we investigated RANK cytoplasmic motifs involved in osteoclast survival. Motif 1, in contrast to its minimal role in osteoclast formation and function, plays a predominant role in promoting osteoclast survival. Moreover, whereas Motif 2 and Motif 3 are highly potent in osteoclast formation and function, they exert a moderate effect on osteoclast survival. We also investigated the role of these motifs in activating Akt/protein kinase B (PKB), which has been implicated in RANKL-induced osteoclast survival. Motif 1, but not Motif 2 or Motif 3, is able to stimulate Akt/PKB activation. Because Akt/PKB has been shown to utilize distinct downstream effectors (glycogen synthase kinase-3beta, FKHR/FOXO1a, BAD, and AFX/FOXO4) to regulate cell survival, we next determined which downstream effector(s) is activated by Akt/PKB to promote osteoclast survival. Our data revealed that RANKL only stimulates AFX/FOXO4 phosphorylation, indicating that AFX/FOXO4 is a key downstream target activated by Akt/PKB to modulate osteoclast survival. Taken together, we conclude that Motif 1 plays a predominant role in mediating osteoclast survival in part by activating Akt/PKB and its downstream effector AFX/FOXO4. PMID- 16260780 TI - Study of highly constitutively active mutants suggests how cAMP activates cAMP receptor protein. AB - The cAMP receptor protein (CRP) of Escherichia coli undergoes a conformational change in response to cAMP binding that allows it to bind specific DNA sequences. Using an in vivo screening method following the simultaneous randomization of the codons at positions 127 and 128 (two C-helix residues of the protein interacting with cAMP), we have isolated a series of novel constitutively active CRP variants. Sequence analysis showed that this group of variants commonly possesses leucine or methionine at position 127 with a beta-branched amino acid at position 128. One specific variant, T127L/S128I CRP, showed extremely high cAMP independent DNA binding affinity comparable with that of cAMP-bound wild-type CRP. Further biochemical analysis of this variant and others revealed that Leu(127) and Ile(128) have different roles in stabilizing the active conformation of CRP in the absence of cAMP. Leu(127) contributes to an improved leucine zipper at the dimer interface, leading to an altered intersubunit interaction in the C helix region. In contrast, Ile(128) stabilizes the proper position of the beta4/beta5 loop by functionally communicating with Leu(61). By analogy, the results suggest two direct local effects of cAMP binding in the course of activating wild-type CRP: (i) C-helix repositioning through direct interaction with Thr(127) and Ser(128) and (ii) the concomitant reorientation of the beta4/beta5 loop. Finally, we also report that elevated expression of T127L/S128I CRP markedly perturbed E. coli growth even in the absence of cAMP, which suggests why comparably active variants have not been described previously. PMID- 16260783 TI - TAK1 is recruited to the tumor necrosis factor-alpha (TNF-alpha) receptor 1 complex in a receptor-interacting protein (RIP)-dependent manner and cooperates with MEKK3 leading to NF-kappaB activation. AB - Receptor-interacting protein (RIP) plays a critical role in tumor necrosis factor alpha (TNF-alpha)-induced IkappaB kinase (IKK) activation and subsequent activation of transcription factor NF-kappaB. However, the molecular mechanism by which RIP mediates TNF-alpha-induced NF-kappaB activation is not completely defined. In this study, we have found that TAK1 is recruited to the TNF-alpha receptor complex in a RIP-dependent manner following the stimulation of TNF-alpha receptor 1 (TNF-R1). Moreover, a forced recruitment of TAK1 to TNF-R1 in the absence of RIP is sufficient to mediate TNF-alpha-induced NF-kappaB activation, indicating that the major function of RIP is to recruit its downstream kinases to the TNF-R1 complex. Interestingly, we also find that TAK1 and MEKK3 form a functional complex, in which TAK1 regulates autophosphorylation of MEKK3. The TAK1-mediated regulation of MEKK3 phosphorylation is dependent on the kinase activity of TAK1. Although TAK1-MEKK3 interaction is not affected by overexpressed TAB1, TAB1 is required for TAK1 activation and subsequent MEKK3 phosphorylation. Together, we conclude that TAK1 is recruited to the TNF-R1 complex via RIP and likely cooperates with MEKK3 to activate NF-kappaB in TNF alpha signaling. PMID- 16260782 TI - Cross-talk between thyroid hormone receptor and liver X receptor regulatory pathways is revealed in a thyroid hormone resistance mouse model. AB - Hypercholesterolemia is found in patients with hypothyroidism and resistance to thyroid hormone. In this study, we examined cholesterol metabolism in a thyroid hormone receptor beta (TR-beta) mutant mouse model of resistance to thyroid hormone. Whereas studies of cholesterol metabolism have been reported in TR-beta knock-out mice, generalized expression of a non-ligand binding TR-beta protein in this knock-in model more fully recapitulates the hypothyroid state, because the hypothyroid effect of TRs is mediated by the unliganded receptor. In the hypothyroid state, a high cholesterol diet increased serum cholesterol levels in wild-type animals (WT) but either did not change or reduced levels in mutant (MUT) mice relative to hypothyroidism alone. 7alpha-Hydroxylase (CYP7A1) is the rate-limiting enzyme in cholesterol metabolism and mRNA levels were undetectable in the hypothyroid state in all animals. triiodothyronine replacement restored CYP7A1 mRNA levels in WT mice but had minimal effect in MUT mice. In contrast, a high cholesterol diet markedly induced CYP7A1 levels in MUT but not WT mice in the hypothyroid state. Elevation of CYP7A1 mRNA levels and reduced hepatic cholesterol content in MUT animals are likely because of cross-talk between TR beta and liver X receptor alpha (LXR-alpha), which both bind to a direct repeat + 4 (DR+4) element in the CYP7A1 promoter. In transfection studies, WT but not MUT TR-beta antagonized induction of this promoter by LXR-alpha. Electromobility shift analysis revealed that LXR/RXR heterodimers bound to the DR+4 element in the presence of MUT but not WT TR-beta. A mechanism for cross-talk, and potential antagonism, between TR-beta and LXR-alpha is proposed. PMID- 16260784 TI - Substrate distortion in the Michaelis complex of Bacillus 1,3-1,4-beta-glucanase. Insight from first principles molecular dynamics simulations. AB - The structure and dynamics of the enzyme-substrate complex of Bacillus 1,3-1,4 beta-glucanase, one of the most active glycoside hydrolases, is investigated by means of Car-Parrinello molecular dynamics simulations (CPMD) combined with force field molecular dynamics (QM/MM CPMD). It is found that the substrate sugar ring located at the -1 subsite adopts a distorted 1S3 skew-boat conformation upon binding to the enzyme. With respect to the undistorted 4C1 chair conformation, the 1S3 skew-boat conformation is characterized by: (a) an increase of charge at the anomeric carbon (C1), (b) an increase of the distance between C1 and the leaving group, and (c) a decrease of the intraring O5-C1 distance. Therefore, our results clearly show that the distorted conformation resembles both structurally and electronically the transition state of the reaction in which the substrate acquires oxocarbenium ion character, and the glycosidic bond is partially broken. Together with analysis of the substrate conformational dynamics, it is concluded that the main determinants of substrate distortion have a structural origin. To fit into the binding pocket, it is necessary that the aglycon leaving group is oriented toward the beta region, and the skew-boat conformation naturally fulfills this premise. Only when the aglycon is removed from the calculation the substrate recovers the all-chair conformation, in agreement with the recent determination of the enzyme product structure. The QM/MM protocol developed here is able to predict the conformational distortion of substrate binding in glycoside hydrolases because it accounts for polarization and charge reorganization at the -1 sugar ring. It thus provides a powerful tool to model E.S complexes for which experimental information is not yet available. PMID- 16260785 TI - The yeast Arr4p ATPase binds the chloride transporter Gef1p when copper is available in the cytosol. AB - Cellular ion homeostasis involves communication between the cytosol and the luminal compartment of organelles. This is particularly critical for metal ions because of their toxic potential. We have identified the yeast homologue of the prokaryotic ArsA protein, the homodimeric ATPase Arr4p, as a protein that binds to the yeast intracellular CLC chloride-transport protein, Gef1p. We show that binding of Arr4p to the C terminus of Gef1p requires the presence of yeast cytosol and is sensitive to a highly specific copper chelator in vitro and in vivo. Copper alone can substitute for cytosol to support the interaction of Arr4p with the C terminus of Gef1p. The migration behavior of Arr4p in nonreducing gel electrophoresis correlates with cellular copper deficiency, repletion, or stress. Our homology model of Arr4p shows that the antimony (arsenic) metal binding site of ArsA is not conserved in Arr4p. The model suggests that a pair of cysteines, Cys285 and Cys288, is located in the interface of the Arr4p dimer. These residues are required for Arr4p homodimerization and for binding to the C terminus of Gef1p. Whereas both proteins are required for normal growth under iron-limiting conditions, they play opposite roles when copper and heat stress are combined in an alkaline environment. Under these conditions, deltagef1 cells grow much better than wild type yeast, whereas deltaarr4 cells are unable to grow. Comparison of the deltaarr4 with the deltaarr4deltagef1 strain suggests that Arr4p antagonizes the function of Gef1p. PMID- 16260786 TI - Molecular basis of the interaction between the flagellar export proteins FliI and FliH from Helicobacter pylori. AB - Bacterial flagellar protein export requires an ATPase, FliI, and presumptive inhibitor, FliH. We have explored the molecular basis for FliI/FliH interaction in the human gastric pathogen Helicobacter pylori. By using bioinformatic and biochemical analyses, we showed that residues 1-18 of FliI very likely form an amphipathic alpha-helix upon interaction with FliH, and that residues 21-91 of FliI resemble the N-terminal oligomerization domain of the F1-ATPase catalytic subunits. A truncated FliI-(2-91) protein was shown to be folded, although the N terminal 18 residues were likely unstructured. Deletion and scanning mutagenesis showed that residues 1-18 of FliI were essential for the FliI/FliH interaction. Scanning mutation of amino acids in the N-terminal 10 residues of FliI indicated that a cluster of hydrophobic residues in this segment was critical for the interaction with FliH. The interaction between FliI and FliH has similarities to the interaction between the N-terminal alpha-helix of the F1-ATPase alpha-subunit and the globular domain of the F1-ATPase delta-subunit, respectively. This similarity suggests that FliH may function as a molecular stator. PMID- 16260788 TI - Familial Parkinson mutant alpha-synuclein causes dopamine neuron dysfunction in transgenic Caenorhabditis elegans. AB - Mutations in alpha-synuclein gene cause familial form of Parkinson disease, and deposition of wild-type alpha-synuclein as Lewy bodies occurs as a hallmark lesion of sporadic Parkinson disease and dementia with Lewy bodies, implicating alpha-synuclein in the pathogenesis of Parkinson disease and related neurodegenerative diseases. Dopamine neurons in substantia nigra are the major site of neurodegeneration associated with alpha-synuclein deposition in Parkinson disease. Here we establish transgenic Caenorhabditis elegans (TG worms) that overexpresses wild-type or familial Parkinson mutant human alpha-synuclein in dopamine neurons. The TG worms exhibit accumulation of alpha-synuclein in the cell bodies and neurites of dopamine neurons, and EGFP labeling of dendrites is often diminished in TG worms expressing familial Parkinson disease-linked A30P or A53T mutant alpha-synuclein, without overt loss of neuronal cell bodies. Notably, TG worms expressing A30P or A53T mutant alpha-synuclein show failure in modulation of locomotory rate in response to food, which has been attributed to the function of dopamine neurons. This behavioral abnormality was accompanied by a reduction in neuronal dopamine content and was treatable by administration of dopamine. These phenotypes were not seen upon expression of beta-synuclein. The present TG worms exhibit dopamine neuron-specific dysfunction caused by accumulation of alpha-synuclein, which would be relevant to the genetic and compound screenings aiming at the elucidation of pathological cascade and therapeutic strategies for Parkinson disease. PMID- 16260787 TI - SHP-2 phosphatase regulates DNA damage-induced apoptosis and G2/M arrest in catalytically dependent and independent manners, respectively. AB - SHP-2, a tyrosine phosphatase implicated in diverse signaling pathways induced by growth factors and cytokines, is also involved in DNA damage-triggered signaling and cellular responses. We previously demonstrated that SHP-2 played an important role in DNA damage-induced apoptosis and G2/M cell cycle checkpoint. In the present studies, we have provided evidence that SHP-2 functions in DNA damage apoptosis and G2/M arrest in catalytically dependent and independent manners, respectively. Mutant embryonic fibroblasts with the Exon 3 deletion mutation in SHP-2 showed decreased apoptosis and diminished G2/M arrest in response to cisplatin treatment. Wild type (WT), but not catalytically inactive mutant SHP-2 (SHP-2 C459S), rescued the apoptotic response of the mutant cells. Interestingly, both WT and SHP-2 C459S efficiently restored the G2/M arrest response. Furthermore, inhibition of the catalytic activity of endogenous SHP-2 in WT cells by overexpression of SHP-2 C459S greatly decreased cell death but not G2/M arrest induced by cisplatin. Biochemical analyses revealed that activation of c-Abl kinase was decreased in SHP-2 C459S-overexpressing cells. However, DNA damage induced translocation of Cdc25C from the nucleus to the cytoplasm was fully restored in both WT and SHP-2 C459S "rescued" cells. Additionally, we demonstrated that the role of SHP-2 in DNA damage-induced cellular responses was independent of the tumor suppressor p53. Embryonic stem cells with the SHP-2 deletion mutation showed markedly decreased sensitivity to cisplatin-induced apoptosis, attributed to impaired induction of p73 but not p53. In agreement with these results, DNA damage-induced apoptosis and G2/M arrest were also decreased in SHP-2/p53 double mutant embryonic fibroblasts. Collectively, these studies have further defined the mechanisms by which SHP-2 phosphatase regulates DNA damage responses. PMID- 16260791 TI - Scientific aspects of ageing: a lordly report. PMID- 16260792 TI - Venous thromboembolism in medical inpatients--the silent epidemic of neglect. PMID- 16260789 TI - Enzymatic redesigning of biologically active heparan sulfate. AB - Heparan sulfate carries a wide range of biological activities, regulating blood coagulation, cell differentiation, and inflammatory responses. The sulfation patterns of the polysaccharide are essential for the biological activities. In this study, we report an enzymatic method for the sulfation of multimilligram amounts of heparan sulfate with specific functions using immobilized sulfotransferases combined with a 3'-phosphoadenosine 5'-phosphosulfate regeneration system. By selecting appropriate enzymatic modification steps, an inactive precursor has been converted to the heparan sulfate having three distinct biological activities, associated with binding to antithrombin, fibroblast growth factor-2, and herpes simplex virus envelope glycoprotein D. Because the recombinant sulfotransferases are expressed in bacteria, and the method uses a low cost sulfo donor, it can be readily utilized to synthesize large quantities of anticoagulant heparin drug or other biologically active heparan sulfates. PMID- 16260794 TI - Embryonic stem cells--sources and destinations. PMID- 16260793 TI - Possession and Jinn. PMID- 16260795 TI - Brain drain from developing countries: how can brain drain be converted into wisdom gain? AB - Brain drain is defined as the migration of health personnel in search of the better standard of living and quality of life, higher salaries, access to advanced technology and more stable political conditions in different places worldwide. This migration of health professionals for better opportunities, both within countries and across international borders, is of growing concern worldwide because of its impact on health systems in developing countries. Why do talented people leave their countries and go abroad? What are the consequences of such migrations especially on the educational sector? What policies can be adopted to stem such movements from developing countries to developed countries? This article seeks to raise questions, identify key issues and provide solutions which would enable immigrant health professionals to share their knowledge, skills and innovative capacities and thereby enhancing the economic development of their countries. PMID- 16260796 TI - Unwanted foreign doctors: what is not being said about the brain drain. PMID- 16260797 TI - The Doctor's PDA and Smartphone Handbook. Personal digital assistant. PMID- 16260798 TI - Research governance: where did it come from, what does it mean? AB - For a variety of historical and social reasons, research has become increasingly formalized and regulated. This change has potential benefits (reduction in fraud and misconduct, protection of vulnerable groups, financial probity) but also disadvantages (increased paperwork, time delays, constraints on research freedom). The terms 'research' and 'governance' mean different things in different contexts. Even with explicit guidance, ambiguities must be resolved by human judgement. Variation in the nature and outcome of approval decisions is therefore a fact of life. The type of approval needed for a research study depends on the official remit of the approval body, the question to be addressed; the methods to be used; the context in which the work will take place; the level of analysis and interpretation; and the plans for how the findings will be presented and used. PMID- 16260799 TI - Palliative care in non-malignant diseases. PMID- 16260800 TI - Venous thromboprophylaxis in UK medical inpatients. AB - We prospectively assessed the implementation of venous thromboembolism (VTE) prophylaxis guidelines and the impact of grand round presentation of the data in changing clinical practice. Two NHS teaching hospitals were studied for 24 months from January 2003. Patients were risk stratified according to the THRIFT (thromboembolic risk factor) consensus group guidelines and compared with the recommendations of the THRIFT and ACCP (American College of Chest Physicians) consensus groups. Six months following presentation of the initial results, a further analysis was made to assess changes in clinical practice. 1128 patients were assessed of whom 1062 satisfied the inclusion criteria for thromboprophylaxis. 89% of all patients were stratified as having high or moderate risk of developing VTE. Of these only 28% were prescribed some form of thromboprophylaxis-4% received the THRIFT-recommended and 22% received the ACCP recommended thromboprophylaxis. The vast majority (72%) received no thromboprophylaxis at all. Reassessment, following data presentation at grand rounds, showed a significant increase to 31% inpatients receiving THRIFT (P<0.0001) and ACCP (P=0.002) recommended thromboprophylaxis. However,the proportion of patients receiving no form of prophylaxis barely changed (72% to 69%: P=0.59). We found a gross underutilization of thromboprophylaxis in hospitalized medical patients. A simple grand-round presentation of the data and recommended guidelines to clinicians significantly increased the proportion of patients receiving recommended thromboprophylaxis but did not increase the overall proportion of patients receiving it. We therefore conclude that a single presentation of guidelines is not enough to achieve the desired levels. Such presentations may only serve to make DVT (deep venous thromboembolism) aware clinicians prescribe prophylaxis more accurately. PMID- 16260801 TI - Cryptococcal small-bowel obstruction in an HIV-positive patient. PMID- 16260802 TI - Pyosalpinx in a man. PMID- 16260804 TI - Treatment with octreotide to suppress corticotropin secretion by a carcinoid tumour. PMID- 16260803 TI - Delayed diagnosis of HIV-associated thrombocytopenia in a man of 70. PMID- 16260808 TI - Lind and scurvy: 1747 to 1795. PMID- 16260809 TI - Challenges to autism research. PMID- 16260810 TI - Now do it our way. PMID- 16260811 TI - Autistic-spectrum disorders: lessons from neuroimaging. AB - Autistic-spectrum disorder is approximately half as common as schizophrenia but its cause remains unknown. Recent studies have begun to clarify the underlying neuroanatomical abnormalities and brain-behaviour relationships in autism. In the past decade, great advances have been made in our understanding of the neurobiological basis of autism. PMID- 16260812 TI - Mind the gap: service transition and interface problems for patients with eating disorders. AB - Recent reports and guidelines that have an impact on the management of people with eating disorders are summarised. The core competencies of every psychiatrist should include: the ability to assess acute risk (including a medical evaluation) and long-term prognosis, and to know what treatments effectively address these needs. PMID- 16260813 TI - Social fragmentation, deprivation and urbanicity: relation to first-admission rates for psychoses. AB - BACKGROUND: Social disorganisation, fragmentation and isolation have long been posited as influencing the rate of psychoses at area level. Measuring such societal constructs is difficult. A census-based index measuring social fragmentation has been proposed. AIMS: To investigate the association between first-admission rates for psychosis and area-based measures of social fragmentation, deprivation and urban/rural index. METHOD: We used indirect standardisation methods and logistic regression models to examine associations of social fragmentation, deprivation and urban/rural categories with first admissions for psychoses in Scotland for the 5-year period 1989-1993. RESULTS: Areas characterised by high social fragmentation had higher first-ever admission rates for psychosis independent of deprivation and urban/rural status. There was a dose-response relationship between social fragmentation category and first-ever admission rates for psychosis. There was no statistically significant interaction between social fragmentation, deprivation and urban/rural index. CONCLUSIONS: First-admission rates are strongly associated with measures of social fragmentation, independent of material deprivation and urban/rural category. PMID- 16260815 TI - Association between stalking victimisation and psychiatric morbidity in a random community sample. AB - BACKGROUND: No studies have assessed psychopathology among victims of stalking who have not sought specialist help. AIMS: To examine the associations between stalking victimisation and psychiatric morbidity in a representative community sample. METHOD: A random community sample (n=1844) completed surveys examining the experience of harassment and current mental health. The 28-item General Health Questionnaire (GHQ-28) and the Impact of Event Scale were used to assess symptomatology in those reporting brief harassment (n=196) or protracted stalking (n=236) and a matched control group reporting no harassment (n=432). RESULTS: Rates of caseness on the GHQ-28 were higher among stalking victims (36.4%) than among controls (19.3%) and victims of brief harassment (21.9%). Psychiatric morbidity did not differ according to the recency of victimisation, with 34.1% of victims meeting caseness criteria 1 year after stalking had ended. CONCLUSIONS: In a significant minority of victims, stalking victimisation is associated with psychiatric morbidity that may persist long after it has ceased. Recognition of the immediate and long-term impacts of stalking is necessary to assist victims and help alleviate distress and long-term disability. PMID- 16260814 TI - IQ and mental disorder in young men. AB - BACKGROUND: Most research investigating the relationship between IQ and risk of mental disorder has focused on schizophrenia. AIMS: To illuminate the relationship between IQ test scores in early adulthood and various mental disorders. METHOD: For 3289 men from the Copenhagen Perinatal Cohort, military IQ test scores and information on psychiatric hospitalisation were available. We identified 350 men in the Danish Psychiatric Central Register, and compared the mean IQ test scores of nine diagnostic categories with the mean scores of 2939 unregistered cohort controls. RESULTS: Schizophrenia and related disorders, other psychotic disorders, adjustment, personality, alcohol and substance-use-related disorders were significantly associated with low IQ scores, but this association remained significant for the four non-psychotic disorders only when adjusting for comorbid diagnoses. For most diagnostic categories, test scores were positively associated with the length of the interval between testing and first admission. ICD mood disorders as well as neuroses and related disorders were not significantly associated with low IQ scores. CONCLUSIONS: Low IQ may be a consequence of mental disease or a causal factor in psychotic and non-psychotic disorders. PMID- 16260816 TI - Heroin dependence in an English town: 33-year follow-up. AB - BACKGROUND: There has been no long-term study of people addicted to injected heroin who have been treated without the prescribing of substitute opioids. AIMS: To investigate the outcome for patients treated for injected heroin addiction 33 years after they were first seen, and 26 years after they were first followed up, in terms of sustained abstinence, continuing maintenance on methadone and deaths. METHOD: Eighty-six people with heroin addiction first seen in in 1966-1967 in a small town in the south-east of England were located and their clinical state assessed using multiple sources, including personal interviews with a proportion of the cohort. RESULTS: Forty-two per cent of the cohort had been abstinent for at least 10 years; 10% were taking methadone and were classified as addicted; and 22% had died. Eight percent of the cohort could not be located. CONCLUSIONS: Results proved favourable in the above three parameters compared with other long term studies. PMID- 16260817 TI - Violent behaviour in schizophrenia. Retrospective study of four independent samples from Prague, 1949 to 2000. AB - BACKGROUND: A number of studies have reported increased violence in patients with schizophrenia. AIMS: To determine the prevalence of violence among those with schizophrenia in samples from 1949, 1969, 1989 and 2000 in Prague (Czech Republic) and to examine trends in this behaviour. METHOD: Records from 404 patients meeting DSM-IV criteria for schizophrenia were screened for violence (defined as 3 points on the Modified Overt Aggression Scale) from the first observed psychotic symptoms until the time of latest available information. RESULTS: Logistic regression revealed a marginally significant increase in violence only in the 2000 cohort. Overall, violence was associated with schizophrenia in 41.8% of men and 32.7% of women, with no association between substance misuse and violence. CONCLUSIONS: The violence rate found in our sample is expected to remain stable over time under stable conditions. Substance misuse is not the leading cause of violence among those with schizophrenia. PMID- 16260819 TI - Processes of disengagement and engagement in assertive outreach patients: qualitative study. AB - BACKGROUND: Assertive outreach has been established to care for'difficult to engage' patients, yet little is known about how patients experience their disengagement with mainstream services and later engagement with outreach teams. AIMS: To explore the views of disengagement and engagement held by patients of assertive outreach teams. METHOD: In-depth interviews were conducted with 40 purposefully selected patients and analysed using components of both thematic analysis and grounded theory. RESULTS: Patients reported a desire to be independent, a poor therapeutic relationship and a loss of control due to medication effects as most important for disengagement. Time and commitment of staff, social support and engagement without a focus on medication, and a partnership model of the therapeutic relationship were most relevant for engagement. CONCLUSIONS: The findings underline the importance of a comprehensive care model, committed staff with sufficient time, and a focus on relationship issues in dealing with 'difficult to engage' patients. PMID- 16260818 TI - Cognitive style in bipolar disorder. AB - BACKGROUND: Abnormalities of cognitive style in bipolar disorder are of both clinical and theoretical importance. AIMS: To compare cognitive style in people with affective disorders and in healthy controls. METHOD: Self-rated questionnaires were administered to 118 individuals with bipolar I disorder, 265 with unipolar major recurrent depression and 268 healthy controls. Those with affective disorder were also interviewed using the Schedules for Clinical Assessment in Neuropsychiatry and case notes were reviewed. RESULTS: Those with bipolar disorder and those with unipolar depression demonstrated different patterns of cognitive style from controls; negative self-esteem best discriminated between those with affective disorders and controls; measures of cognitive style were substantially affected by current levels of depressive symptomatology; patterns of cognitive style were similar in bipolar and unipolar disorder when current mental state was taken into account. CONCLUSIONS: Those with affective disorder significantly differed from controls on measures of cognitive style but there were no differences between unipolar and bipolar disorders when current mental state was taken into account. PMID- 16260820 TI - Experience of caring for someone with anorexia nervosa: qualitative study. AB - BACKGROUND: Caring for someone with anorexia nervosa is distressing. AIMS: To gain a detailed understanding of carers' illness models and caregiving experiences. METHOD: Qualitative analysis and computerised text analysis were conducted on narratives written by parents as part of a family intervention at a specialist in-patient unit (20 mothers, 20 fathers). RESULTS: Themes concerned illness perceptions, impact on the family, and carers'emotional, cognitive and behavioural responses towards the illness. Parents perceived anorexia nervosa to be chronic and disabling. Carers blamed themselves as contributing to the illness and perceived themselves as helpless in promoting recovery. Mothers illustrated an intense emotional response, whereas fathers produced a more cognitive and detached account. CONCLUSIONS: Part of the distress in living with anorexia nervosa may be explained by unhelpful assumptions and maladaptive responses to the illness. Training parents in skills to manage the illness may improve outcome by reducing interpersonal maintaining factors. PMID- 16260821 TI - Reality orientation therapy combined with cholinesterase inhibitors in Alzheimer's disease: randomised controlled trial. AB - BACKGROUND: Reality orientation therapy combined with cholinesterase inhibitors has not been evaluated in patients with Alzheimer's disease. AIMS: To perform such an evaluation. METHOD: We randomly assigned 79 of 156 patients treated with donepezil to receive a reality orientation programme. Caregivers of the treatment group were trained to offer the programme at home 3 days a week, 30 min/day, for 25 consecutive weeks, and were invited to stimulate and involve patients in reality-based communication. RESULTS: The treatment group showed a slight improvement in Mini-Mental State Examination (MMSE) scores (mean change +0.2, s.e.=0.4) compared with a decline in the control group (mean change -1.1, s.e.=0.4; P=0.02). Similarly for the Alzheimer's Disease Assessment Scale- Cognition (treatment group mean change +0.4, s.e.=0.8; control group -2.5, s.e.=0.8; P=0.01). The intervention had an equal effect on cognition in those with mild (MMSE score > or = 20) and moderate (score <20) dementia. No significant effect was observed for behavioural and functional outcomes. CONCLUSIONS: Reality orientation enhances the effects of donepezil on cognition in Alzheimer's disease. PMID- 16260822 TI - Internet-based self-help for depression: randomised controlled trial. AB - BACKGROUND: Major depression can be treated by means of cognitive-behavioural therapy, but as skilled therapists are in short supply there is a need for self help approaches. Many individuals with depression use the internet for discussion of symptoms and to share their experience. AIMS: To investigate the effects of an internet-administered self-help programme including participation in a monitored, web-based discussion group, compared with participation in web-based discussion group only. METHOD: A randomised controlled trial was conducted to compare the effects of internet-based cognitive-behavioural therapy with minimal therapist contact (plus participation in a discussion group) with the effects of participation in a discussion group only. RESULTS: Internet-based therapy with minimal therapist contact, combined with activity in a discussion group, resulted in greater reductions of depressive symptoms compared with activity in a discussion group only (waiting-list control group). At 6 months' follow-up, improvement was maintained to a large extent. CONCLUSIONS: Internet-delivered cognitive cognitive-behavioural therapy should be pursued further as a complement or treatment alternative for mild-to-moderate depression. PMID- 16260823 TI - Adherence to guidelines for treatment of depression in in-patients. AB - BACKGROUND: Adherence to treatment guidelines enhances treatment outcome. However, in clinical practice many patients with depression do not receive appropriate treatment. AIMS: To evaluate the treatment of depression in in patients of German psychiatric hospitals with respect to treatment outcome and adherence to guidelines. METHOD: We recruited 1202 in-patients with depression from ten different hospitals. Quality data concerning treatment were collected at admission, during the treatment course and at discharge. RESULTS: The level of depression was significantly decreased and most patients were satisfied with treatment. Many aspects of the treatment routine adhered to guideline recommendations. Adherence to guidelines could be improved with respect to adjustment of antidepressant dosage, reduction of benzodiazepine prescription, enhanced use of electroconvulsive therapy and wider use of interpersonal therapy. CONCLUSIONS: The study reveals a high standard of psychiatric treatment of in patients with depression. Nevertheless there is still room for improvement. Differences between hospitals in adherence to guidelines indicates the need for individual application of quality management tools. PMID- 16260824 TI - Mortality and suicide after non-fatal self-poisoning: 16-year outcome study. AB - BACKGROUND: Suicide reduction is government strategy in many countries. We need to quantify the connection between non-fatal self-poisoning and eventual suicide. AIMS: To determine mortality after an episode of self-poisoning and to identify predictors of death by any cause or by suicide. METHOD: A retrospective single group cohort study was undertaken with 976 consecutive patients attending a large accident and emergency unit in 1985-1986 after non-fatal self-poisoning. Information about deaths was determined from the Office for National Statistics. RESULTS: Of the original patients, 94% were traced 16 years later; 17% had died, 3.5% by probable suicide. Subsequent suicide was related to numerous factors evident at the time of the episode of self-poisoning but, when examined for their independent effects, only the severity of the self-poisoning episode and relevant previous history seemed important. CONCLUSIONS: Patients attending a general hospital after self-poisoning all require good basic assessment and care responsive to their needs. Attempts to reduce the huge excess of suicide subsequent to self-harm are not likely to achieve much if they are based on the identification of subgroups through'risk assessment'. PMID- 16260825 TI - Clustering of suicides among people with mental illness. AB - BACKGROUND: Most previous investigations of imitative suicide have reported suicide clustering in the general population, either temporal clustering following media reporting of suicide or case studies of geographically localised clusters. AIMS: To determine whether space - time and space-time-method clustering occur in a national case register of those who had recent contact with mental health services and had died by suicide and to estimate the suicide imitation rate in this population. METHOD: Knox tests were used for space-time and space-time-method clustering. Model simulations were used to estimate effect size. RESULTS: Highly significant space-time and space-time-method clustering was found in a sample of 2741 people who died by suicide over 4 years who had had recent contact with one of 105 mental health trusts. Model simulations with an imitation rate of 10.1% (CI 4-17) reproduced the observed space-time-method clustering. CONCLUSIONS: This study provides indirect evidence that imitative suicide occurs among people with mental illnesses and may account for about 10% of suicides by current and recent patients. PMID- 16260827 TI - A role for psychedelics in psychiatry? PMID- 16260826 TI - Structural brain correlates of unconstrained motor activity in people with schizophrenia. AB - Avolition affects quality of life in chronic schizophrenia. We investigated the relationship between unconstrained motor activity and the volume of key executive brain regions in 16 male patients with schizophrenia. Wristworn actigraphy monitors were used to record motor activity over a 20 h period. Structural magnetic resonance imaging brain scans were parcellated and individual volumes for anterior cingulate cortex and dorsolateral prefrontal cortex extracted. Patients'total activity was positively correlated with volume of left anterior cingulate cortex. These data suggest that the volume of specific executive structures may affect (quantifiable) motor behaviours, having further implications for models of the 'will' and avolition. PMID- 16260828 TI - A role for psychedelics in psychiatry? PMID- 16260829 TI - A role for psychedelics in psychiatry? PMID- 16260830 TI - Olanzapine co-therapy in bipolar disorder. PMID- 16260831 TI - CBT for refractory symptoms in schizophrenia. PMID- 16260832 TI - ECT in depression. PMID- 16260833 TI - Hospital admission rates and diagnosis. PMID- 16260834 TI - Zero tolerance of violence. PMID- 16260835 TI - Structure and anticipatory movements of the S6 gate in Kv channels. PMID- 16260836 TI - Status of the intracellular gate in the activated-not-open state of shaker K+ channels. AB - Voltage-dependent K+ channels like Shaker use an intracellular gate to control ion flow through the pore. When the membrane voltage becomes more positive, these channels traverse a series of closed conformations before the final opening transition. Does the intracellular gate undergo conformational changes before channel opening? To answer this question we introduced cysteines into the intracellular end of the pore and studied their chemical modification in conditions favoring each of three distinct states, the open state, the resting closed state, and the activated-not-open state (the closed state adjacent to the open state). We used two independent ways to isolate the channels in the activated-not-open state. First, we used mutations in S4 (ILT; Smith-Maxwell, C.J., J.L. Ledwell, and R.W. Aldrich. 1998. J. Gen. Physiol. 111:421-439; Ledwell, J.L., and R.W. Aldrich. 1999. J. Gen. Physiol. 113:389-414) that separate the final opening step from earlier charge-movement steps. Second, we used the open channel blocker 4-aminopyridine (4-AP), which has been proposed to promote closure of the intracellular gate and thus specifically to stabilize the activated-not-open state of the channels. Supporting this proposed mechanism, we found that 4-AP enters channels only after opening, remaining trapped in closed channels, and that in the open state it competes with tetraethylammonium for binding. Using these tools, we found that in the activated-not-open state, a cysteine located at a position considered to form part of the gate (Shaker 478) showed higher reactivity than in either the open or the resting closed states. Additionally, we have found that in this activated state the intracellular gate continued to prevent access to the pore by molecules as small as Cd2+ ions. Our results suggest that the intracellular opening to the pore undergoes some rearrangements in the transition from the resting closed state to the activated not-open state, but throughout this process the intracellular gate remains an effective barrier to the movement of potassium ions through the pore. PMID- 16260837 TI - Generation of functional fluorescent BK channels by random insertion of GFP variants. AB - The yellow and cyan variants of green fluorescent protein (GFP) constitute an excellent pair for fluorescence resonance energy transfer (FRET) and can be used to study conformational rearrangements of proteins. Our aim was to develop a library of fluorescent large conductance voltage- and Ca2+-gated channels (BK or slo channels) for future use in FRET studies. We report the results of a random insertion of YFP and CFP into multiple sites of the alpha subunit of the hslo channel using a Tn5 transposon-based technique. 55 unique fluorescent fusion proteins were obtained and tested for cell surface expression and channel function. 19 constructs are expressed at the plasma membrane and show voltage and Ca2+-dependent currents. In 16 of them the voltage and Ca2+ dependence is very similar to the wild-type channel. Two insertions in the Ca2+ bowl and one in the RCK2 domain showed a strong shift in the G-V curve. The remaining 36 constructs were retained intracellularly; a solubility assay suggests that these proteins are not forming intracellular aggregates. The "success rate" of 19 out of 55 hslo insertion constructs compares very favorably with other studies of random GFP fusions. PMID- 16260838 TI - A spatiotemporal white noise analysis of photoreceptor responses to UV and green light in the dragonfly median ocellus. AB - Adult dragonflies augment their compound eyes with three simple eyes known as the dorsal ocelli. While the ocellar system is known to mediate stabilizing head reflexes during flight, the ability of the ocellar retina to dynamically resolve the environment is unknown. For the first time, we directly measured the angular sensitivities of the photoreceptors of the dragonfly median (middle) ocellus. We performed a second-order Wiener Kernel analysis of intracellular recordings of light-adapted photoreceptors. These were stimulated with one-dimensional horizontal or vertical patterns of concurrent UV and green light with different contrast levels and at different ambient temperatures. The photoreceptors were found to have anisotropic receptive fields with vertical and horizontal acceptance angles of 15 degrees and 28 degrees, respectively. The first-order (linear) temporal kernels contained significant undershoots whose amplitudes are invariant under changes in the contrast of the stimulus but significantly reduced at higher temperatures. The second-order kernels showed evidence of two distinct nonlinear components: a fast acting self-facilitation, which is dominant in the UV, followed by delayed self- and cross-inhibition of UV and green light responses. No facilitatory interactions between the UV and green light were found, indicating that facilitation of the green and UV responses occurs in isolated compartments. Inhibition between UV and green stimuli was present, indicating that inhibition occurs at a common point in the UV and green response pathways. We present a nonlinear cascade model (NLN) with initial stages consisting of separate UV and green pathways. Each pathway contains a fast facilitating nonlinearity coupled to a linear response. The linear response is described by an extended log-normal model, accounting for the phasic component. The final nonlinearity is composed of self-inhibition in the UV and green pathways and inhibition between these pathways. The model can largely predict the response of the photoreceptors to UV and green light. PMID- 16260840 TI - Calcium activation of ryanodine receptor channels--reconciling RyR gating models with tetrameric channel structure. AB - Despite its importance and abundance of experimental data, the molecular mechanism of RyR2 activation by calcium is poorly understood. Recent experimental studies involving coexpression of wild-type (WT) RyR2 together with a RyR2 mutant deficient in calcium-dependent activation (Li, P., and S.R. Chen. 2001. J. Gen. Physiol. 118:33-44) revealed large variations of calcium sensitivity of the RyR tetramers with their monomer composition. Together with previous results on kinetics of Ca activation (Zahradnikova, A., I. Zahradnik, I. Gyorke, and S. Gyorke. 1999. J. Gen. Physiol. 114:787-798), these data represent benchmarks for construction and testing of RyR models that would reproduce RyR behavior and be structurally realistic as well. Here we present a theoretical study of the effects of RyR monomer substitution by a calcium-insensitive mutant on the calcium dependence of RyR activation. Three published models of tetrameric RyR channels were used either directly or after adaptation to provide allosteric regulation. Additionally, two alternative RyR models with Ca binding sites created jointly by the monomers were developed. The models were modified for description of channels composed of WT and mutant monomers. The parameters of the models were optimized to provide the best approximation of published experimental data. For reproducing the observed calcium dependence of RyR tetramers containing mutant monomers (a) single, independent Ca binding sites on each monomer were preferable to shared binding sites; (b) allosteric models were preferable to linear models; (c) in the WT channel, probability of opening to states containing a Ca2+-free monomer had to be extremely low; and (d) models with fully Ca-bound closed states, additional to those of an Monod-Wyman-Changeaux model, were preferable to models without such states. These results provide support for the concept that RyR activation is possible (albeit vanishingly small in WT channels) in the absence of Ca2+ binding. They also suggest further avenues toward understanding RyR gating. PMID- 16260841 TI - Coupled K+-water flux through the HERG potassium channel measured by an osmotic pulse method. AB - The streaming potential (V(stream)) is a signature feature of ion channels in which permeating ions and water molecules move in a single file. V(stream) provides a quantitative measure of the ion and water flux (the water-ion coupling ratio), the knowledge of which is a prerequisite for elucidating the mechanisms of ion permeation. We have developed a method to measure V(stream) with the whole cell patch-clamp configuration. A HEK293 cell stably expressing the HERG potassium channel was voltage clamped and exposed to hyperosmotic solutions for short periods of time (<1 s) by an ultrafast solution switching system (the osmotic pulse [quick jump-and-away] method). The reversal potentials were monitored by a series of voltage ramps before, during, and after the osmotic pulse. The shifts of the reversal potentials immediately after the osmotic jump gave V(stream). In symmetrical K+ solutions (10 mM), the V(stream)s measured at different osmolalities showed a linear relationship with a slope of -0.7 mV/DeltaOsm, from which the water-ion coupling ratio (n, the ratio of the flux of water to the flux of cations; Levitt, D.G., S.R. Elias, and J.M. Hautman. 1978. Biochim. Biophys. Acta. 512:436-451) was calculated to be 1.4. In symmetrical 100 mM K+ solutions, the coupling ratio was decreased significantly (n = 0.9), indicating that the permeation process through states with increased ion occupancy became significant. We presented a diagrammatic representation linking the water-ion coupling ratio to the mode of ion permeation and suggested that the coupling ratio of one may represent the least hydrated ion flux in the single file pore. PMID- 16260839 TI - Charge screening by internal pH and polyvalent cations as a mechanism for activation, inhibition, and rundown of TRPM7/MIC channels. AB - The Mg2+-inhibited cation (MIC) current, believed to represent activity of TRPM7 channels, is found in lymphocytes and mast cells, cardiac and smooth muscle, and several other eukaryotic cell types. MIC current is activated during whole-cell dialysis with divalent-free internal solutions. Millimolar concentrations of intracellular Mg2+ (or other divalent metal cations) inhibit the channels in a voltage-independent manner. The nature of divalent inhibition and the mechanism of channel activation in an intact cell remain unknown. We show that the polyamines (spermine, spermidine, and putrescine) inhibit the MIC current, also in a voltage-independent manner, with a potency that parallels the number of charges. Neomycin and poly-lysine also potently inhibited MIC current in the absence of Mg2+. These same positively charged ions inhibited IRK1 current in parallel with MIC current, suggesting that they probably act by screening the head group phosphates on PIP2 and other membrane phospholipids. In agreement with this hypothesis, internal protons also inhibited MIC current. By contrast, tetramethylammonium, tetraethylammonium, and hexamethonium produced voltage dependent block but no inhibition. We show that inhibition by internal polyvalent cations can be relieved by alkalinizing the cytosol using externally applied ammonium or by increasing pH in inside-out patches. Furthermore, in perforated patch and cell-attached recordings, when intracellular Mg2+ is not depleted, endogenous MIC or recombinant TRPM7 currents are activated by cytosolic alkalinization and inhibited by acidification; and they can be reactivated by PIP2 following rundown in inside-out patches. We propose that MIC (TRPM7) channels are regulated by a charge screening mechanism and may function as sensors of intracellular pH. PMID- 16260842 TI - Potential method of optimizing the klapskate hinge position in speed skating. AB - Acceptance of the klap speed skate was fully realized on the world speed skating scene in 1997. However, one of the most important unknowns regarding the klapskate was the positioning of the point of foot rotation (pivot point), which is believed to play an important role in optimizing klapskate performance. The purposes of this study were to explore the ankle, knee, and hip joint mechanical changes that occurred when the pivot point location was modified, and to determine whether maximal ankle torques provide predictive ability as to where the optimal pivot point positioning is for a skater. We tested 16 proficient skaters at three pivot point PP) locations, ranging from just in front of the metatarsal-phalangeal joint to just in front of the first phalangeal joint. Of the 16 skaters, 10 were tested at a fourth position; tip of the toe. Push phase kinetics and kinematics were measured on a modified slide board. The optimal PP for each skater was defined as the position that allowed him to generate the most total push energy. Maximum voluntary static torque measures of the ankle and knee were collected on a Biodex dynamometer. Overall, anterior pivot point shifting led to a significant increase in ankle energy generated and a decrease in knee energy generated, with no significant change at the hip joint. We found no significant correlations between the static strength measures and the skaters' optimal pivot points. PMID- 16260843 TI - The margin for error when releasing the asymmetric bars for dismounts. AB - It has previously been shown that male gymnasts using the "scooped" giant circling technique were able to flatten the path followed by their mass center, resulting in a larger margin for error when releasing the high bar (Hiley and Yeadon, 2003a). The circling technique prior to performing double layout somersault dismounts from the asymmetric bars in women's artistic gymnastics appears to be similar to the "traditional" technique used by some male gymnasts on the high bar. It was speculated that as a result the female gymnasts would have margins for error similar to those of male gymnasts who use the traditional technique. However, it is unclear how the technique of the female gymnasts is affected by the need to avoid the lower bar. A 4-segment planar simulation model of the gymnast and upper bar was used to determine the margins for error when releasing the bar for 9 double layout somersault dismounts at the Sydney 2000 Olympics. The elastic properties of the gymnast and bar were modeled using damped linear springs. Model parameters, primarily the inertia and spring parameters, were optimized to obtain a close match between simulated and actual performances in terms of rotation angle (1.2 degrees), bar displacement (0.011 m), and release velocities (<1%). Each matching simulation was used to determine the time window around the actual point of release for which the model had appropriate release parameters to complete the dismount successfully. The margins for error of the 9 female gymnasts (release window 43-102 ms) were comparable to those of the 3 male gymnasts using the traditional technique (release window 79-84 ms). PMID- 16260844 TI - Vertical jumping reorganization with aging: a kinematic comparison between young and elderly men. AB - To our knowledge jumping kinematics have never been studied in elderly persons. This study was aimed at examining the influence of aging on vertical jump performance and on interjoint coordination. Two groups of adults, 11 young men ages 18-25 years and 11 older men ages 79-100 years, were filmed while performing a maximal squat jump. Compared to young adults, jump height was significantly decreased by 28 cm in the elderly. Older adults spontaneously jumped from a more extended position of the hip. Results showed a decrease in hip, knee, and ankle linear velocity and angular amplitude with aging. The decrease in jump height was attributed to a decrease in explosive force and in the range of shortening of extensor muscles. In agreement with the literature, a proximo-distal coordination pattern was observed in young adults. Older adults used a simultaneous pattern. This may indicate that adults adjust their pattern of joint coordination as they age. PMID- 16260845 TI - Dynamics of the in-run in ski jumping: a simulation study. AB - A ski jumper tries to maintain an aerodynamic position in the in-run during changing environmental forces. The purpose of this study was to analyze the mechanical demands on a ski jumper taking the in-run in a static position. We simulated the in-run in ski jumping with a 4-segment forward dynamic model (foot, leg, thigh, and upper body). The curved path of the in-run was used as kinematic constraint, and drag, lift, and snow friction were incorporated. Drag and snow friction created a forward rotating moment that had to be counteracted by a plantar flexion moment and caused the line of action of the normal force to pass anteriorly to the center of mass continuously. The normal force increased from 0.88 G on the first straight to 1.65 G in the curve. The required knee joint moment increased more because of an altered center of pressure. During the transition from the straight to the curve there was a rapid forward shift of the center of pressure under the foot, reflecting a short but high angular acceleration. Because unrealistically high rates of change of moment are required, an athlete cannot do this without changing body configuration which reduces the required rate of moment changes. PMID- 16260846 TI - The effect of a repetitive, fatiguing lifting task on horizontal ground reaction forces. AB - There are many outdoor work environments that involve the combination of repetitive, fatiguing lifting tasks and less-than-optimal footing (muddy/slippery ground surfaces). The focus of the current research was to evaluate the effects of lifting-induced fatigue of the low back extensors on lifting kinematics and ground reaction forces. Ten participants performed a repetitive lifting task over a period of 8 minutes. As they performed this task, the ground reaction forces and whole body kinematics were captured using a force platform and magnetic motion tracking system, respectively. Fatigue was verified in this experiment by documenting a decrease in the median frequency of the bilateral erector spinae muscles (pretest-posttest). Results indicate significant (p < 0.05) increases in the magnitude of the peak anterior/posterior (increased by an average of 18.3%) and peak lateral shear forces (increased by an average of 24.3%) with increasing time into the lifting bout. These results have implications for work environments such as agriculture and construction, where poor footing conditions and requirements for considerable manual materials handling may interact to create an occupational scenario with an exceptionally high risk of a slip and fall. PMID- 16260847 TI - Filtering of kinematic signals using the Hodrick-Prescott filter. AB - The use of the Hodrick-Prescott (HP) filter is presented as an alternative to the traditional digital filtering and spline smoothing methods currently used in biomechanics. In econometrics, HP filtering is a standard tool used to decompose a macroeconomic time series into a nonstationary trend component and a stationary residual component. The use of the HP filter in the present work is based on reasonable assumptions about the jerk and noise components of the raw displacement signal. Its applicability was tested on 4 kinematic signals with different characteristics. Two are well known signals taken from the literature on biomechanical signal filtering, and the other two were acquired with our own motion capture system. The criterion for the selection of cutoff frequency was based on the power spectral density of the raw displacement signals. The results showed the technique to be well suited to filtering biomechanical displacement signals in order to obtain accurate higher derivatives in a simple and systematic way. Namely, the HP filter and the generalized cross-validated quintic spline (GCVSPL) produce similar RMS errors on the first (0.1063 vs. 0.1024 m/s2) and second (23.76 vs. 23.24 rad/s2) signals. The HP filter performs slightly better than GCVSPL on the third (0.209 vs. 0.236 m/s2) and fourth (1.596 vs. 2.315 m/s2) signals. PMID- 16260848 TI - The influence of stretcher height on the mechanical effectiveness of rowing. AB - The aim of the present study was to determine the effect of varying the height of the foot stretcher on the mechanical effectiveness of rowing. Ten male university level rowers rowed maximally for 3 minutes 30 seconds on a modified Concept 2 rowing ergometer. Each participant completed one trial at three foot stretcher heights. Position 1 was the original Concept 2 stretcher position, with Position 2 being located 5 cm and Position 3 being 10 cm above the original position and in the same orientation. Pull force and velocity were measured, and mean power generated by the rowers was calculated for each stroke. It was shown that in all three stretcher positions, mean power per stroke decreased as a function of time during the trial, confirming the fatiguing effects of the task. Although mean power per stroke did not differ significantly between stretcher positions at the start of the trial, p = 0.082, a significant difference was observed between the original stretcher position and Positions 2 and 3 at the end of the trial, p < 0.05. The lowest decline in mean power occurred in the highest stretcher position. It is suggested that this improvement in effectiveness is due to a reduction in the active downward vertical forces applied to the foot stretchers which does not contribute to forward propulsion, and thus a reduction in energy waste during each stroke. It was hypothesized that further raising the stretchers will continue to lead to an improvement in effectiveness until the optimum stretcher height is reached, above which effectiveness will be reduced. PMID- 16260849 TI - Modification of cycling biomechanics during a swim-to-cycle trial. AB - The aim of this study was to investigate the effects of drafting, i.e., swimming directly behind a competitor, on biomechanical adaptation during subsequent cycling. Eight well-trained male triathletes underwent three submaximal sessions in a counterbalanced order. These sessions comprised a 10-min ride on a bicycle ergometer at 75% of maximal aerobic power (MAP) at a freely chosen cadence. This exercise was preceded either by a 750-m swim performed alone at competition pace (SCA trial; swimming-cycling alone), a 750-m swim in a drafting position at the same pace as during SCA (SCD trial; swimming-cycling with drafting), or a cycling warm-up at 30% of MAP for the same duration as the SCA trial (CTRL trial). The results indicated that the decrease in metabolic load when swimming in a drafting position (SCD trial) was associated with a significantly lower pedal rate and significantly higher mean and peak resultant torques when compared to the SCA trial, p < 0.05. These results could be partly explained by the lower relative intensity during swimming in the SCD trial when compared with the SCA trial, involving a delayed manifestation of fatigue in the muscles of the lower limbs at the onset of cycling. PMID- 16260851 TI - Adjuvant chemotherapy in 780 patients with early breast cancer: 10-year data from Saudi Arabia. AB - BACKGROUND: By and large, data about adjuvant chemotherapy for breast cancer in the Middle East are lacking. Retrospective analysis of prospectively captured data from a main referral center in the Kingdom of Saudi Arabia (KSA) may shed some light on the clinicopathological features and survival of patients offered adjuvant chemotherapy in a similar population in that part of the world. PATIENTS AND METHODS: Data on patients with invasive breast cancer (Stages I to IIIA) seen between 1992 and the end of 2001 and who received adjuvant chemotherapy were analyzed. A total of 780 patients were considered eligible and constitute the basis of this report. RESULTS: The median age +/- SD of the 780 patients was 42 +/- 9.6 yr. The majority of patients were younger than 50 yr (78%) and premenopausal (83%). Ten percent, 69%, and 21% of patients had Stage I, II, and IIIA, respectively. Patients expressed relatively high prevalence of adverse clinicopathological characteristics. Most patients (523 patients, 67%) received anthracyclines-containing adjuvant chemotherapy, 610 patients (78%) received adjuvant radiotherapy, and 296 (38%) received adjuvant tamoxifen. At a median follow-up of 42 mo (95% CI, 38.1-62.8 mo), the median overall (OS) and disease free survival (DFS) were not reached; however, the 5-yr actuarial survival was estimated as 74% and 59%, respectively. Cox proportional regression hazard model identified positive axillary nodal status, and positive vascular invasion are the only variables that influenced OS adversely. The model also distinguished the same variables plus negative estrogen receptor status as covariates with negative effect on DFS. CONCLUSION: In conclusion, this series of 780 predominantly young patients with breast cancer receiving adjuvant chemotherapy highlighted the disease patterns and survival outcome in the KSA. The current series is significant being one of the few reports about adjuvant chemotherapy experience in a developing country and certainly the first from that part of the world. PMID- 16260850 TI - Immunoglobulin gene analysis of mature B-cell malignancies: reconsideration of cellular origin and potential antigen involvement in pathogenesis. AB - Mature B-cell malignancies stem from B cells transformed at various developmental stages, accounting for the wide range of heterogeneous features observed in the different disease entities. Analysis of the immunoglobulin (Ig) genes can facilitate the identification of the normal B-cell counterpart of lymphomas and leukemias, as Ig genes acquire somatic hypermutation in germinal centers during the immune response to antigen. Therefore, lymphomas that derive from a naive, pregerminal center B cell lack somatic hypermutation in the clonal Ig gene, whereas germinal center-derived lymphomas, such as diffuse large B-cell lymphoma and follicular lymphoma, display somatic hypermutation of their Ig genes. Furthermore, biases in the Ig variable heavy chain gene repertoire in B-cell malignancies can indicate a possible antigenic influence in pathogenesis. Much work has been accomplished in the past decade to characterize the Ig genes in different lymphoma entities, and the separation of chronic lymphocytic leukemia into two prognostic subgroups in the late 1990s based on the presence or absence of somatic hypermutation led to investigations of Ig genes in larger cohorts of previously uncharacterized entities, such as mantle cell lymphoma. This review will briefly discuss relevant aspects of normal B-cell development, and then focus on what can be ascertained from Ig studies of newly characterized entities, mantle cell lymphoma, hairy cell leukemia, lymphoplasmacytic lymphoma/ Waldenstrom's macroglobulinemia, and splenic marginal zone lymphoma, from the point of view of cellular origin and variable heavy chain gene restrictions as a sign of antigen involvement. Correlations with gene expression profiling data and the clinical implications of Ig gene studies, when relevant, will be mentioned. The recent evidence that an alternative pathway of gaining somatic hypermutation might exist is also considered, and the implications this has for understanding the cellular origin of B-cell malignancies. PMID- 16260852 TI - Serum levels of leptin and proinflammatory cytokines in advanced-stage non-small cell lung cancer. AB - In this study, we aimed to investigate the diagnostic and prognostic roles and the effects of chemotherapy of serum proinflammatory cytokines consisting of IL 6, TNF-alpha, CRP, and leptin levels in patients with advanced-stage non-small cell lung cancer. Twenty-eight patients newly diagnosed of non-surgical advanced non-small cell lung cancer and 15 healthy controls were included. All patients with good performance status were treated with combination therapy consisting of cisplatin plus vinorelbine chemotherapy. Blood samples were obtained in fasting conditions before chemotherapy first and after two cycles of chemotherapy. IL-6 and TNF-alpha immunoassays employ the quantitative sandwich enzyme immunoassay technique. Leptin (Sandwich) ELISA is a solid-phase enzyme-linked immunosorbent assay based on the sandwich principle. CRP is a photometric immunoturbidimetric test. Most of the patients were elderly, male predominance, good performance status, and no or less than 10% weight loss. Higher serum TNF-alpha (p < 0.001) and CRP (p < 0.001), and lower leptin (p = 0.021) levels in patients than in controls. Serum IL-6 cytokine (p = 0.693) levels were not significantly different. No statistically significant relationships between investigated serum parameters and various characteristics of patient and disease. Likewise, serum levels of leptin, IL-6, TNF-alpha, and CRP were all similar in lung cancer patients independently from severity of weight loss (p > 0.05). A direct relationship was found between serum IL-6 and TNF-alpha levels (r = 0.530, p = 0.004). We found that both serum leptin (p = 0.046) and IL-6 (p = 0.002) levels were decreased owing to the chemotherapy effect independently from chemotherapy response. However, serum TNF-alpha and CRP levels were not changed by the chemotherapy effect. The stage of the disease, serum LDH levels, performance status, and responsiveness to chemotherapy yielded prognostic value. Only serum IL-6 levels out of the parameters showed a trend (p = 0.06) related to a worse prognosis. PMID- 16260853 TI - Gemcitabine plus carboplatin in patients with advanced non-small cell lung cancer. AB - To evaluate the efficacy and safety of gemcitabine combined with carboplatin for a chemotherapy regimen for patients with metastatic, recurrent, or locally advanced non-small cell lung cancer (NSCLC), 46 chemotherapy-naive patients with histologically confirmed stage IIIB or IV NSCLC were treated with 1250 mg/m(2) of gemcitabine on d 1 and 8, with carboplatin of AUC 6 additionally applied on d 1. This treatment was repeated every 3 wk. In all, a total of 215 chemotherapy courses were administered. The median age of the patients was 46, ranging from 33 to 83. Ten patients (22%) had an ECOG performance status of 2. Responses were observed objectively in 20 patients (43.5%) and maintained for a median of 7.4 mo. The median duration of progression-free and overall survivals were 5.0 and 12.3 mo, respectively. Neutropenia was frequently encountered, and gastrointestinal side effects, such as anorexia and nausea, were mostly predictable but manageable. One patient died of septic shock due to a complication with pneumonia while simultaneously trying to recover from myelosuppression. A subgroup consisting of patients aged 65 yr or older and/or PS 2 showed a outcome similar with the entire group of all patients involved in the study: response rate (43.5%), median PFS (4.6 months), median OS (12.3 months), and similar toxicity rate. After analyzing all the results, it was evident that a treatment of gemcitabine combined with carboplatin is an active and safe regimen for first-line treatment of advanced NSCLC. The results of the elderly and/or PS 2 patients were similar to those of the entire group of patients. PMID- 16260855 TI - Role of gross tumor volume on outcome and of dose parameters on toxicity of patients undergoing chemoradiotherapy for locally advanced non-small cell lung cancer. AB - The aim of this retrospective study was to evaluate the prognostic role of gross tumor volume (GTV) on survival of locally advanced NSCLC patients, regardless of TNM stage, and to analyze whether GTV and other radiotherapy (RT) parameters were important for the development of lung toxicity. Thirty-two patients with locally advanced NSCLC (stage IIIA bulky/IIIB) treated with chemoradiotherapy were retrospectively analyzed. Patients received induction chemotherapy followed by combination treatment (27 patients) or induction chemotherapy followed by RT alone (5 patients). Thoracic RT consisted in 60 Gy, with standard fractionation and was the same for all 32 patients. Dose volume histograms were collected from the 3D treatment plans and GTV, planning target volume, mean lung dose, volume of lung receiving more than 20 Gy or more than 30 Gy were analyzed. Survival was significantly longer in patients with a GTV < 100 cm(3) compared with patients having GTV > 100 cm(3) (p = 0.03). In a multivariate analysis only N-status and GTV were predictors of survival with a risk ratio of 0.51 and 0.62, respectively. Ten patients (31%) developed radiation pneumonitis grade 2 or higher. None of the RT parameters examined correlated significantly with the development of lung toxicity. In locally advanced NSCLC, GTV and N-status play a prognostic role even in patients at the same clinical stage and receiving a combination of chemo- and radiotherapy. This could imply a reassessment of the current staging system in patients with non-resectable NSCLC to better identify those patients who would benefit more from the combined treatment, despite its higher toxicity. PMID- 16260854 TI - Induction and concurrent chemotherapy with concomitant boost radiotherapy in non small cell lung cancer. AB - This study was designed to evaluate the tolerability and therapeutic activity of paclitaxel and carboplatin combination therapy followed by radical thoracic radiotherapy with a concomitant boost technique with concurrent weekly paclitaxel in good performance status of patients with stage IIIA and IIIB non-small cell lung cancer. Patients with newly diagnosed inoperable non-small cell lung cancer received paclitaxel (100 mg/m(2)) as a 1-h infusion on d 1,8,15,28,35, and 42. Carboplatin (area under the curve of 6) was given as a 30-min infusion on d 1 and 28. Radiotherapy commenced on d 49 and was delivered with accelerated fractionation with concomitant boost at 1.8 Gy/fraction/d, 5 d/week and 1.5 Gy/fraction/d to a boost field as a second daily treatment for the last 10 treatment days to 60 Gy/35 fractions/5 wk. During radiation treatment, paclitaxel (60 mg/m(2)) was given as a 1-h infusion once weekly for 5 wk. Twenty-four patients were enrolled in the study. Hematologic toxicities and alopecia were the major acute toxicities during induction chemotherapy; 8.7% of the patients experienced grade 3-4 neutropenia and alopecia. The main acute toxicity of concurrent chemoradiotherapy was esophagitis; grade 3 esophagitis was documented in 23.5% of the patients. No major late toxicity was seen. Overall response rate to the treatment was 65.2%. The median and 1-yr overall-survival rates were 24.9 mo and 63.8%, respectively. The median and 1-yr progression-free survival rates were 9.0 mo and 27.8%, respectively. The main acute toxicities were hematologic toxicity, esophagitis, and alopecia. The response rate and the survival rates achieved with this treatment regimen are particularly noteworthy, especially considering the advanced stage of the patients treated. PMID- 16260857 TI - Overexpression of cyclooxygenase-2 in gastric cancer correlates with the high abundance of vascular endothelial growth factor-C and lymphatic metastasis. AB - Cyclooxygenase-2 (COX-2) and vascular endothelial growth factor (VEGF)-C are closely related with the development and metastasis of tumors. The gene expression of COX-2 and VEGF-C in gastric cancer and the correlation between them were investigated; 64 paraffin-embedded gastric cancer samples and 22 flesh gastric cancer samples were tested by using immunohistochemistry and the reverse transcription polymerase chain reaction (RT-PCR) technology, respectively. The mean expressive density of COX-2 and VEGF-C mRNA in gastric cancer, with beta actin coamplified as an internal standard, were both significantly higher than those in non-cancerous gastric mucosa (1.363 +/- 0.351 vs 0.763 +/- 0.304, 0.972 +/- 0.331 vs 0.314 +/- 0.215, p < 0.001). The positive rates of COX-2 and VEGF-C in 64 gastric cancer samples were 72% and 64% respectively. Their expression in the lymph-node metastasis groups were higher than that of the non-lymph-node metastasis groups (p < 0.05). Moreover, there was a close correlation between COX 2 and VEGF-C expression levels (p < 0.05). The study indicates gastric tumor tissues that produce COX-2 and VEGF-C may have a higher lymphatic invasion and metastatic potential. COX-2 may participate in VEGF-C lymphangiogenic pathway and the high expression of them may play an important role in the lymphatic proliferation and spread in gastric cancer. PMID- 16260856 TI - Residual disease after chemotherapy in aggressive malignant lymphoma: the role of radiotherapy. AB - Residual disease in patients with diffuse large B-cell lymphoma after intensive chemotherapy remains a problem. Radiotherapy has been used in some retrospective studies without definitive conclusions. We report the first controlled clinical trial to define the role of radiotherapy in this setting of patients. One hundred and sixty-six patients with diagnosis of diffuse large B-cell lymphoma, high- or high-intermediate clinical risk, with residual disease (defined as tumor mass < 5 cm) were randomly assigned to received radiotherapy at the involved field, with 30 Gy delivered in 20 sessions or no radiation (control group). Median follow-up was 135 mo; patients who received radiotherapy have an better outcome. Actuarial curves at 10 yr showed that progressive-free disease was 86% and overall survival was 89%; those were statistical significant when compared to patients who did no received radiotherapy: 32% and 58% respectively, (p < 0.001). Toxicity was mild and well tolerated. We concluded that presence of residual mass after chemotherapy in patients with aggressive malignant lymphoma has a worse prognosis, and salvage radiotherapy improves outcome with mild toxicity. We feel that radiotherapy will be considered as necessary treatment in this special group of patients. PMID- 16260858 TI - Implications of therapy choice on overall survival in metastatic renal cell carcinoma: a single institution experience. AB - PURPOSE: To compare the effect of high-dose interleukin-2 (HD IL-2) vs other cytokine therapies on 1-, 2, and 5-yr overall survival in patients with metastatic renal cell cancer (RCC). PATIENTS AND METHODS: We conducted a retrospective chart review of patients with untreated metastatic RCC treated by a single investigator. The different treatment groups included HD IL-2, low-dose IL 2 alone or in combination, interferon alpha alone and other therapies. The primary end point was survival from time of treatment. RESULTS: A total of 85 patients were studies with a median follow up of 13 mo (0.6-112.9). Median age at treatment was 59 yr with predominantly male patients and histology of clear cell type. Thirty-four percent received HD IL-2 and treatment was initiated less than 6 mo from the time of diagnosis in 66%. For all patients, median survival was 16 mo with a 5-yr survival of 12%. Two factors were good predictors of overall survival: Karnofsky performance status (KPS) of 100 (p < 0.0001) and soft tissue metastasis (p = 0.01). When comparing HD IL-2 to all other groups, median survival was 18 vs 14 mo and 1-yr survival was 74% vs 51%, respectively. CONCLUSION: HD IL-2 is associated with clinically meaningful improvement in median and 1-yr survival. PMID- 16260859 TI - Spontaneous remission in adults with primary myelodysplastic syndromes: incidence and characteristics of patients. AB - Reports on spontaneous remissions in patients with primary myelodysplastic syndromes (MDS) occasionally appear in the literature. We report five adult patients with spontaneous remission of MDS, achieved without cytotoxic or any other treatment. These five patients represent 1.6% of 307 MDS patients, diagnosed in our Institute since 1987. According to FAB criteria, three patients had RA, and one patient had RARS and RAEB, each. All patients were women, median age of 63 yr (range 32-68 yr). Patients were without significant complaints and peripheral cytopenia was mild. Bone marrow dyshematopoiesis was also mild, mostly affecting erythroid and megakaryocytic series. At diagnosis, three patients had cytogenetic abnormalities [+8,+12; +15 and del(16)(q22)]. Median time to complete hematological and cytogenetical remission was 51 mo, while median duration of spontaneous remission was 45 mo (range 44-60 mo). As for the follow-up, none of the patients relapsed. In conclusion, although spontaneous remissions (i.e., "regression") of MDS are uncommon, better understanding of their basis may lead to crucial advances in the study of leukemogenesis. PMID- 16260860 TI - Clonal populations of chondrocytes with progenitor properties identified within human articular cartilage. AB - The aim of the present study was to identify and characterize progenitor properties of human articular chondrocytes selected by using agarose suspension culture. In this chondrogenic selective culture condition, about 3.6% of seeded surplus chondrocytes from patients undergoing articular chondrocyte transplantation proliferated and formed cell clusters after 6 weeks. Phase contrast microscopy and transmission electron microscopy revealed four different types of cell clusters differing in cellular content and matrix production. Based on their morphological features, they were named the homogenous (H), the homogenous matrix (HM), the differentiated matrix (DM) and the differentiated (D) cell clusters. All cell clusters showed positive safranin O staining, and matrix was positive for antibodies detecting type II collagen and aggrecan. The clusters were further demonstrated to express the genes for fibroblast growth factor receptor 3, type IIA collagen and type IIB collagen, while type X collagen was not expressed. After subcloning, the H and HM clusters demonstrated the best proliferative capacity. Chondrocytes from these two cell clusters also showed phenotypic plasticity in chondrogenic, adipogenic as well as osteogenic assays. This study demonstrates that existing subpopulations of cells with chondroprogenitor properties can be isolated from human adult articular cartilage using agarose suspension cultures. PMID- 16260861 TI - Low oxygen tension enhances chondroinduction by demineralized bone matrix in human dermal fibroblasts in vitro. AB - Endochondral bone formation is induced by demineralized bone powder (DBP) when DBP is implanted subcutaneously in rodents. Previously, we developed an in vitro model of this process, wherein human dermal fibroblasts (hDFs) differentiate to chondrocytes when cultured in a three-dimensional porous collagen sponge containing DBP. In other studies, medium perfusion was beneficial in maintaining phenotype and viability of many cell types in plain porous collagen sponges, including fibroblasts, bone marrow stromal cells, osteoblasts, and epidermal cells. In contrast, medium perfusion inhibited chondrogenesis by articular chondrocytes; reduction of oxygen tension to 5%, however, restored chondrogenesis. These observations are consistent with the fact that in vivo cartilage is avascular and relatively hypoxic compared with other vascularized tissues. In this study, we tested the hypothesis that low oxygen tension (hypoxia, 5% oxygen) would enhance induced chondrogenesis in hDFs cultured with DBP. As expected, hypoxia upregulated hypoxia-inducible factor-1alpha in hDFs in all conditions (i.e. +/- perfusion, +/- DBP). Hypoxia increased accumulation of cartilage-specific matrix chondroitin 4-sulfate in hDFs, but only in the presence of DBP (165%, compared to normoxia, p < 0.05). Hypoxia did not appear to have detrimental effects on cell viability and proliferation. In sum, hypoxia enhanced cartilage matrix accumulation by hDFs cultured with DBP. These defined conditions can optimize the use of dermal fibroblasts for cartilage tissue engineering. PMID- 16260862 TI - Rhodopsin, violet and blue opsin expressions in the chick are highly dependent on tissue and serum conditions. AB - The molecular, cellular or tissue environment can influence the expression of genes and thereby regulate processes of tissue formation. Here we determined the tissue and serum dependence of the expression of all photopigments in the chick by a series of distinct retinal cell cultures, analyzed by RT-PCR using specific primers for all four opsins and rhodopsin followed by quantitative scanning of the respective gel bands. For comparison, we first determined expression of all opsins during normal chick retinogenesis, which began with red and violet opsins at E12, shortly followed by blue and green opsins and finally rhodopsin at E14. This period corresponds to the time of synaptogenesis in the inner retina. All cultures were started with 6-day-old dissociated retinal cells. Cells were kept at low or high cell density (called LoDens or HiDens), or they were reaggregated as retinal spheres, whereby all of them were raised at low (2%) or high serum (12%) levels (called LoSer or HiSer). In LoDens/HiSer cultures, expression of all opsins was weak. At HiDens/LoSer red and green opsin expression was strong, while rhodopsin and violet/blue remained low. In HiDens/HiSer cultures the expression of red and green was strong; rhodopsin was almost normal, while violet and green were low. In reaggregates at high serum the expression came closest to a normal retina, but violet and blue opsins were still below normal. At low serum, however, violet and blue were negligible and rhodopsin was low. This in vitro study shows that rhodopsin, followed by violet and blue opsin expressions is highly dependent on serum, cell density and tissue conditions, while red and green opsins are more autonomous. PMID- 16260863 TI - Vectorial secretion of perivitelline membrane glycoprotein ZPC of Japanese quail (Coturnix japonica) in polarized Madin-Darby canine kidney cells. AB - The avian perivitelline membrane (PL), which is an investment homologous to the mammalian zona pellucida, is found between the surface of the oocyte and the apical surface of ovarian granulosa cells. Our previous study demonstrated that ZPC, one of the components of PL, is synthesized in ovarian granulosa cells. However, how the secretion of ZPC is regulated in the cells has been insufficiently investigated. We studied the secretion of quail ZPC expressed in polarized Madin-Darby canine kidney (MDCK) cells in a dual-chamber apparatus. Western blot analyses of the conditioned medium demonstrated that the majority of the secreted ZPC were distributed in the apical compartment. When ZPC lacking N linked oligosaccharides was transfected into the cells, the 31-kDa immunoreactive band was detected in both the apical and the basolateral medium. Interestingly, immunohistochemical observations of the follicular wall demonstrated that the predominant intracellular form of ZPC in the cells localized in the apical side of the perinuclear region apposed to the PL, but not the basolateral side, indicating the possibility that ZPC could be selectively transported toward the apical surface in vivo. Taken together, these results indicated that ZPC expressed in MDCK cells are selectively released to the apical compartment, and that the N-linked carbohydrates might possess information that causes the efficient transport of ZPC to the apical surface of the cells. PMID- 16260864 TI - Constitutive coexpression of nitric oxide synthase-1 and soluble guanylyl cyclase in myoepithelial cells of mammary glands in mice. AB - An impact of nitric oxide (NO) on lactation and milk secretion in mammary glands has previously been documented, but the underlying molecular mechanisms for this modulatory effect remain unclear. Therefore, we investigated the expression patterns of NO synthase (NOS)-1, NOS-3 and the NO receptor soluble guanylyl cyclase (sGC) in mammary glands of lactating and non-lactating female C57/Bl6 mice. RT-PCR demonstrated the existence of NOS-1-mRNA and NOS-3-mRNA in both lactating and resting mammary tissue. Immunoblots loaded with equal amounts of homogenate proteins from lactating and resting mammary tissues revealed comparable intensities of NOS-1 and sGC bands. Performing catalytic NADPH diaphorase histochemistry and immunohistochemistry, NOS-1 was only detected in myoepithelial cells (MEC), while sGC was localized in alveolar epithelial cells (lactocytes) and MEC in both lactating and non-lactating mammary glands. The non modulated co-expression of both enzymes suggests that NOS-1 and sGC contribute to the constitutive regulation of tone in MEC. PMID- 16260865 TI - Age-related morphological changes in the human hyoid bone. AB - OBJECT: Age-related morphological changes in the human hyoid bone were investigated radiographically and histologically. MATERIALS AND METHODS: Thirty two measurements were performed on radiographs of 238 hyoid bones from autopsy cases of known age and sex. Thirty-one hyoid bones that were studied by radiography were also examined histologically in horizontal sections. RESULTS: Analysis of the length and width of the hyoid bone revealed significant increases in the body and the anterior part of the greater cornu and a significant decrease in the posterior part of the greater cornu with aging. Most measurements of the body and the greater cornu revealed differences between male and female hyoid bones. The outer margins of the body and the greater cornu were situated further outside in older males compared with females. The breadth of the joint space showed a significant age-related decrease, and the degree of fusion showed a significant age-related increase. Histological findings showed ossified or calcified fusion, with osteoclasts in the marginal area of the joint space. CONCLUSIONS: Increasing age induces fusion of the body and the greater cornu. The morphometric changes in the shape of the hyoid bone may represent functional adaptation to articulation fixation. PMID- 16260866 TI - Clinical features of chronic pancreatitis in Korea: a multicenter nationwide study. AB - BACKGROUND/AIMS: No reliable nationwide clinical data about chronic pancreatitis (CP) was available in Korea. The etiology and clinical features of CP were investigated using a multicenter nationwide study. METHODS: 814 cases of CP were enrolled retrospectively over the past 4 years at 13 hospitals. The following data were obtained from all patients: etiology, symptoms, complications, and surgery. RESULT: Alcohol (64.3%) was the major cause of CP and idiopathic CP (20.8%) was the second most common form. Mean patient age was 50.6 years and the male:female ratio was 6:1 (24:1 for alcoholic CP vs. 2:1 in idiopathic CP, p < 0.001). Diabetes (31.6%), pseudocysts (28.4%), biliary stricture (13.9%), and pancreatic ascites (6.6%) were the main complications. Of these, diabetes (35 vs. 26%) and pseudocyst (33.7 vs. 21.9%) were more frequent in alcoholic than in idiopathic CP. Pancreatic cancer developed in 25 patients (3.1%) during follow-up and their mean age was 59.1 years. CONCLUSIONS: In Korea, alcohol is the most common etiology of CP. Moreover, diabetes and pseudocysts are frequent complications, especially in alcoholic CP, and pancreatic cancer development is not infrequent. PMID- 16260868 TI - Bilateral video-assisted thoracoscopic thymectomy vs. extended transsternal thymectomy in myasthenia gravis: a prospective study. AB - The optimal approach to thymectomy remains controversial. This study is designed to prospectively compare the results between bilateral video-assisted thoracoscopic thymectomy (BVTx) and extended transsternal thymectomy (ETTx) in patients with myasthenia gravis (MG) without thymoma. Fifteen patients who had undergone BVTx and 16 patients who had undergone ETTx were compared for age, gender, severity of disease, preoperative duration of disease, operative time, intraoperative blood loss, postoperative complications, hospital stay, duration of chest tube drainage, thymic histopathology, pain perception by visual analog scale (VAS), remission and improvement rate, period of follow-up, and activities of daily living (ADL). Fisher's exact test, t test and paired t test were used for statistical analysis. BVTx had longer operative time and less intraoperative blood loss than that of the ETTx. Their remission rates and their degree of postoperative ADL improvement were not significantly different. However, the lowering of VAS was significantly greater in the sternotomy group at 3 months. All other parameters were not significantly different. No mortality was noted in the series. We consider BVTx as an effective alternative procedure to the transsternal approach for patients with nonthymomatous MG. As more and more people care about cosmetics, BVTx could become the future trend. PMID- 16260867 TI - Combination of enoxaparin and fibroblast growth factor-1 increases myocardial blood flow and capillary density after myocardial infarction in rabbits. AB - OBJECTIVE: The effect of enoxaparin and fibroblast growth factor-1 (FGF-1) on post-infarction capillary density and regional myocardial blood flow (RMBF) was examined. METHODS: New Zealand White rabbits received an intramyocardial injection of either physiological saline, FGF-1 + enoxaparin, FGF-1 or enoxaparin directly after ligation of the left anterior descending artery. RMBF and capillary density were investigated using fluorescent microspheres and histological examination. RESULTS: One week after infarction a significant difference in the number of capillaries could be demonstrated within the FGF-1 + enoxaparin group (p < 0.001 versus the control group), the FGF-1 group (p < 0.01) and the enoxaparin group (p < 0.05). Treatment with FGF-1 + enoxaparin resulted in a significantly increased number of capillaries compared to treatment with FGF 1 (p < 0.05) and enoxaparin (p < 0.05) alone. Additionally, all groups treated with FGF-1 and/or enoxaparin showed a significant increase of microvessel density in the treated ischemic border zone compared to the non-treated ischemic border zone (p < 0.001 for FGF-1 + enoxaparin, p < 0.01 for FGF-1, p < 0.05 for enoxaparin). RMBF was significantly increased within the FGF-1 + enoxaparin group compared to the control group (p < 0.05). Moreover, perfusion rates within the FGF-1 + enoxaparin-treated area did not significantly differ from the pre infarction values. CONCLUSION: Treatment with either enoxaparin or FGF-1 or FGF-1 + enoxaparin resulted in increased microvessel growth. However, only the combination of enoxaparin with FGF-1 promotes capillary growth and RMBF. Thus, we conclude that enoxaparin enhances the angiogenic potential of intramyocardially injected FGF-1 in the acutely infarcted rabbit heart. PMID- 16260869 TI - Development of a special balloon occlusion device to prevent adverse events in high-risk patients during open aortic surgery. AB - OBJECTIVE: To prevent clamp injury that may occur during aortic surgery, we aimed to develop a special balloon occlusion (BO) device to lower the thromboembolic risk in patients with severe atherosclerosis during aortic aneurysm repair. METHODS: The study comprised two test phases: a laboratory-testing series focussing on flexible artificial aortas, and an experimental series conducted on 10 pigs. RESULTS: The device proved to be effective during the laboratory tests and the experiments on pigs. No complications such as intraoperative balloon rupture, dislocation, or occlusion leaks occurred. No damage to the aortic vessels was observed in further histological examinations. CONCLUSIONS: This BO device has the potential to become an alternative to cross-clamping for vascular surgeons in patients with severely atherosclerotic vessels. PMID- 16260871 TI - D-dimer in the early diagnosis of acute mesenteric ischemia secondary to arterial occlusion in rats. AB - BACKGROUND: Early diagnosis of mesenteric ischemia can be life saving. The aim of this study is to investigate the time-dependent diagnostic value of plasma D dimer and other ancillary laboratory evaluations such as creatine phosphokinase, lactic dehydrogenase, aspartate aminotransferase, alanine aminotransferase, amylase, and leukocyte count in an experimental superior mesenteric arterial occlusion (SMA-O) model in rats. METHODS: Forty male Wistar-Albino rats were separated into 4 groups: 2-, 4-, 6-, and 12-hour SMA-O groups. After laparotomy, the SMA was identified and ligated for 2, 4, 6 and 12 h in the 4 respective study groups. Blood samples were taken for laboratory tests 2 h after laparotomy in the control group and at the end of the ischemic period in the study groups. RESULTS: The longer the duration of mesenteric ischemia, the higher were the serum D-dimer levels in the study groups, and statistical significance was obtained at 2 h (p = 0.021). Sensitivity, positive predictive value, negative predictive value, and accuracy of the relation were 88.8, 88.8, 100 and 90%, respectively. Leukocyte count was significantly higher than controls after 12 h. No other laboratory parameter correlated positively with the duration of mesenteric ischemia. CONCLUSION: Serum D-dimer measurements may be a valuable diagnostic parameter in the early diagnosis of mesenteric ischemia. PMID- 16260870 TI - Reduced capacity of whole blood leucocytes to express tumour necrosis factor alpha and interleukin-10 following major orthopaedic surgery. AB - BACKGROUND: Severe trauma is a challenge to the immune response and may cause reduced immune capacity. As a marker of decreased cellular activity, studies with ex vivo lipopolysaccharide (LPS) stimulation of whole blood or isolated mononuclear cells from injured patients have revealed reduced production of inflammatory cytokines. To gain further insight into immune alterations in orthopaedic surgery, we studied LPS-induced tumour necrosis factor (TNF)-alpha and interleukin (IL)-10 in whole blood of patients during peri- and postoperative phases of total hip replacement. METHODS: Four females and 3 males undergoing elective total hip replacement were included in the study. Ex vivo LPS-induced TNF-alpha and IL-10 were measured in a whole blood assay before, during and at 1 and 6 days after operation. In addition, the counts of white blood cells were determined. RESULTS: During the operation, there were significant reductions in the number of monocytes, but at day 1 and 6 after surgery, there were significant increases as compared to the levels before surgery. The capacity of whole blood to express TNF-alpha and IL-10 did not change significantly during the operation and the following postoperative day. At day 6, however, there were significant reductions in expression of both TNF-alpha and IL-10 as compared to the levels before the operation. In relation to the values of monocytes, there was a significant reduction in the expression of TNF-alpha also at day 1 after operation. CONCLUSION: Our data indicate that in the course of at least 6 days after a major orthopaedic trauma, there is suppression of the whole blood capacity to express the inflammatory cytokine TNF-alpha and the anti-inflammatory cytokine IL-10 when exposed to LPS. During this time, then, the patient is particular susceptible to septic complications. PMID- 16260873 TI - Irradiated pancreatic cancer cells undergo both apoptosis and necrosis, and could be phagocytized by dendritic cells. AB - The interaction of immature dendritic cells (DC) with irradiated pancreatic cancer cells was examined. Flow cytometric analysis using annexin V and propidium iodide revealed that ionizing radiation (25-35 Gy X-ray) induced both apoptosis and necrosis in pancreatic cancer cell lines. After irradiation, PK-1 and Panc-1 cells were likely to undergo necrosis, whereas MIAPaCa-2 cells underwent apoptosis. When DiO-stained immature DCs were co-incubated with DiI-stained irradiated MIAPaCa-2, it was observed under fluorescent microscopy that DCs phagocytized dead tumor cells as early as 4 h after co-incubation. The DCs' phagocytosis of irradiated tumor cells was also confirmed by flow cytometry. These results suggest that irradiated pancreatic cancer cells, which undergo both apoptosis and necrosis, could be a good source of tumor-associated antigens for cross-presentation by DCs. PMID- 16260872 TI - Interleukin-10 gene transfection of donor pancreas grafts protects against rejection after heterotopic pancreas transplantation in a rat model. AB - OBJECTIVE: The aim of this study was to assess the effect of immunoregulatory cytokine interleukin-10 (IL-10) gene therapy on pancreas tissue rejection in a heterotopic pancreas transplantation model. BACKGROUND: Modulation of inflammatory responses by anti-inflammatory cytokines (e.g., IL-10) has been suggested to minimize organ rejection. In this context, modulation of cytokines using gene therapy could be a new therapeutic modality in preventing organ rejection. METHODS: The study was performed using male inbred Wistar rats as recipients and Sprague-Dawley rats as donors. 24 h before transplantation, groups of rats, named IL-10 (n = 20) and green fluorescent protein (GFP, n = 20), were injected with viral vectors Ad5CMVhIL10 or Ad5CMVGFP. Sham-operated rats (n = 20) underwent saline injection only before transplantation. The pancreatic tissue from each of these donor rats was subsequently transplanted into the corresponding groups of streptozotocin-induced diabetic recipient rats. Recipients were thus transfected with either IL-10 (n = 20), GFP-only carrying viral vectors (n = 20) or no viral vectors (normal saline, n = 20). A selected number of animals from each recipient group (n = 5) was sacrificed at weekly intervals for 3 weeks and some were further followed up to 12 weeks before sacrifice. Histological assessment of the pancreatic tissue was made based on rejection and GFP expression. Blood glucose levels were checked daily in all groups until sacrifice. Upon sacrifice, serum cytokine and insulin levels were measured. Histopathological correlations between blood glucose levels, serum insulin levels and serum IL-10 levels were made. RESULTS: IL-10 gene therapy significantly attenuated pancreas rejection compared to controls, provided more normal blood glucose levels and elevated plasma insulin levels. Upon assumed natural deactivation of transferred viruses after 4 weeks, differences between groups in terms of rejection, blood glucose and insulin levels disappeared. CONCLUSION: These findings suggest that IL-10 gene therapy significantly reduced pancreas rejection. PMID- 16260874 TI - Coincidence of connective tissue growth factor expression with fibrosis and angiogenesis in postoperative peritoneal adhesion formation. AB - PURPOSE: To investigate the relationship between connective tissue growth factor (CTGF) and fibrosis and angiogenesis in postoperative peritoneal adhesion formation. METHODS: Adhesions were performed in 35 rats by creation of a peritoneal patch. Animals were sacrificed at 7 different time-points over 3 weeks. Adhesions and uninjured peritoneum from all animals were assessed by Northern blotting for CTGF and collagen-I mRNA and by immunohistochemistry for CTGF localization, degree of fibrosis and angiogenesis. RESULTS: Persistent adhesions formed in all animals. CTGF and collagen-I mRNA were increased in adhesions compared to uninjured peritoneum (p < 0.05 for both). The temporal expression pattern depicted delayed peak levels of collagen-I mRNA with increasing tendency for both transcripts at the end of the observation period. Fibrosis within adhesions correlated positively with time after surgery (r = 0.85; p < 0.001) and showed typical signs of chronic tissue fibrosis at later time points. Angiogenesis was detected in adhesions but not in uninjured peritoneum (p = 0.001) and coincided with the spatial and temporal expression of CTGF protein in fibroblasts and vascular endothelial cells. CONCLUSIONS: The co expression of CTGF with increasing fibrosis and angiogenesis in postoperative peritoneal adhesions suggests a role for CTGF as critical molecule in fibrous adhesive disease and target for future adhesion prevention. PMID- 16260875 TI - Effect of povidone-iodine liposome hydrogel on colonic anastomosis. AB - BACKGROUND: To evaluate the effect of povidone-iodine liposome hydrogel on colonic anastomosis. METHODS: 70 Wistar-Albino male rats were randomly divided into seven groups. The left colon was transected and end-to-end anastomosis was performed. PVP-I liposome hydrogel was applied around the anastomoses in groups 2 and 5. Colonic bursting pressures and tissue hydroxyproline contents were measured on postoperative days 3 and 7. RESULTS: PVP-I application did not cause any difference in the bursting pressures on postoperative day 3, but anastomotic strength was significantly increased by the use of PVP-I on postoperative day 7. The use of PVP-I liposome hydrogel had no effect on the level of perianastomotic hydroxyproline on postoperative day 3 but had positive effects on postoperativeday 7. CONCLUSIONS: We concluded that the beneficial effects of PVP I liposome hydrogel might be due to the combination of broad-spectrum antimicrobial activity of PVP-I with the drug delivery properties and moisturizing molecular film effects of the liposome hydrogel. PMID- 16260876 TI - Methylene blue in the evaluation of gastrointestinal tract integrity: potential limitations. AB - INTRODUCTION: The integrity of the gastrointestinal tract can be evaluated by oral or rectal application of methylene blue. In the presence of anastomotic leaks or fistulas, methylene blue can be recovered in adjacent drains. However, parts of the dye can biochemically be reduced by intestinal bacteria to its colorless form leucomethylene blue, limiting the prediction of the test. MATERIALS AND METHODS: Diluted methylene blue was added to different concentrations of bacterial suspensions of Escherichia coli and Enterococcus faecalis. The time for discoloration of the suspension was measured. RESULTS: Reduction of methylene blue to leucomethylene blue was observed in both types of bacterial suspension. A 10(8) bacterial concentration discolorated the dye within 1 h in the E. faecalis suspension, respectively 2.5 h in the E. coli suspension. Longer bacterial interaction with methylene blue reduced the bacterial concentration required to achieve complete discoloration. DISCUSSION: Methylene blue can reliably be used as routine diagnostic test for the assessment of upper gastrointestinal integrity, where bacterial load is low. In the lower gastrointestinal tract, where bacterial load is generally higher, the dye can indicate leaks, only if extended intestinal passage after oral ingestion is avoided. In all other cases, the examiner has to be aware of false-negative results by bacterial discoloration of methylene blue. PMID- 16260877 TI - Effect of pantoprazole administered subcutaneously on the healing of sutured gastric incisions in rats. AB - BACKGROUND: The properties of proton pump inhibitors most investigated are related to peptic diseases and upper gastrointestinal bleeding, but their influence on the healing of sutured gastric incisions has not been assessed. In the present study we evaluated the effect of subcutaneously administered pantoprazole on the healing of sutured gastric incisions in rats. METHODS: Sixty rats were divided into a pantoprazole group and a control group. All rats were submitted to gastric suture in the gastric body and in the gastric fundus and had their gastric pH measured. The pantoprazole group received 20 mg/kg pantoprazole and the control group received 0.9% isotonic NaCl, both subcutaneously t.i.d. Healing analysis was carried out using collagen picrosirius red F3BA staining, and breaking strength was measured on the 4th and 7th postoperative days in all groups. RESULTS: Gastric pH was higher in the pantoprazole group. In the fundus, the pantoprazole group had a higher measurement of breaking strength and a higher proportion of type-I over type-III collagen on the 7th postoperative day. In the body, the pantoprazole group had a higher proportion of type-I over type-III collagen on the 4th and 7th postoperative days. CONCLUSIONS: Pantoprazole given subcutaneously promoted a reduction in gastric acid secretion and was associated with improved healing of the sutured gastric incision in the fundus (squamous epithelium) of rats. These findings suggest that pantoprazole has healing properties in sutured gastric incisions with potential benefits in gastric surgery. PMID- 16260878 TI - Testicular-epididymal hemodynamics and inguinal hernia. AB - Testicular-epididymal hemodynamics is studied by ultrasonographic color velocity imaging (CVI) in patients with primary, uncomplicated inguinal hernias. This procedure allows more precision than any other known Doppler system in measuring hemodynamic parameters such as peak systolic and diastolic velocity, resistance index and pulsatility index. The results of the exploration of 480 arteries of various testicular vascular territories are compared with those obtained by other authors in young healthy individuals using color Doppler. This procedure shows that the hernia does not cause significant alterations in the arterial circulation of the testicle and epididymis. PMID- 16260879 TI - Elevated aldosterone in amniotic fluid and maternal blood has diagnostic potential in pregnancies complicated with a fetus of Bartter syndrome. AB - Pregnancies with fetuses affected with the Bartter syndrome, an autosomal recessive disorder of hyperreninism and hyperaldosteronism, are complicated by early onset of polyhydramnios which results in preterm deliveries. We have assessed biochemical changes in amniotic fluid and the mother's blood with a view to early diagnosis. Aldosterone levels of both amniotic fluid and the mother's blood were found to be increased at 27 weeks of gestation, while electrolyte levels did not differ significantly from those reported earlier for controls. After birth the baby suffered from polyuria with hyponatremia, hypomagnesemia and hypercalciuria which could be controlled by treatment with sodium chloride and magnesium. Elevated aldosterone thus might be a useful marker for early diagnostic purposes. PMID- 16260880 TI - Feasibility of retroviral vector-mediated in utero gene transfer to the fetal rabbit. AB - OBJECTIVES: Successful treatment or prevention of severe hereditary diseases could conceivably be achieved by genetic intervention early in development. Viral vector-mediated fetal gene transfer is proving a valuable tool to test the above concept in relevant animal models. Although the pregnant rabbit is a well recognized model for fetal therapy, few preclinical assays have used it to validate fetal gene transfer approaches. In this preliminary study we assessed for the first time the feasibility of retroviral vector-mediated in utero gene transfer in the fetal rabbit. METHODS: Different amounts of the vesicular stomatitis virus G pseudotyped MFG(nls)LacZ retroviral vector, expressing a nuclear-localized beta-galactosidase reporter protein were injected intraperitoneally and -hepatically into 20- to 22-day-old fetuses. At 8-9 days post-treatment, the pups were sacrificed and the tissues harvested for analysis. Evidence of gene transfer was obtained by PCR amplification of proviral sequences within genomic DNA isolated from the treated samples. Transgenic beta galactosidase expression was assessed by X-gal histochemical staining. RESULTS: By intraperitoneal injection 43% of the viable fetuses treated (3/7) showed evidence of successful LacZ gene transfer and low-level beta-galactosidase expression into liver and heart, whereas by intrahepatic injection roughly 38% (3/8) of the livers were positive for LacZ gene transfer and expression. The success rate for the viable fetuses rose to 67% positive livers (4/6) when a near double amount of recombinant virus was injected using a 10-fold concentrated virus stock. In terms of short-term safety, fetal and maternal survival rates approached 80% of treated fetuses, and 100% of treated does. CONCLUSIONS: The pregnant rabbit is a useful and reliable model allowing the design of further studies to optimize the conditions for effective, safer, and persistent retroviral vector-mediated fetal gene transfer. PMID- 16260881 TI - Aminiotic fluid and intrauterine growth restriction in a gastroschisis fetal rat model. AB - Fetuses with digestive anomalies such as gastroschisis may present intrauterine growth restriction (IUGR) and shortened intestines. OBJECTIVE: The aim of this study was to assess the influence caused by amniotic fluid (AF) in intestinal length and somatic growth in an experimental gastroschisis fetal model at two distinct gestational ages. MATERIAL AND METHOD: Fetal rats were operated according to Correia-Pinto on 2 different days of gestation: day 18.5 (group I) and day 19.5 (group II). Each group was divided into three sub-groups: fetuses with gastroschisis (G), control (C) and sham(S). Body measurements and histological analysis were done. RESULT: Body measurement analysis showed: average body weight (g) in group I was G = 5.32, C = 5.68, S = 5.86; group II was G = 5.32, C = 5.80, S = 5.66. Average intestine weight (g) in group I was G = 0.283, C = 0.238, S = 0.231; group II was G = 0.272, C = 0.231, S = 0.233. Average intestine length (mm) in group I was G = 125, C = 216, S = 209; group II was G = 148, C = 226, S = 226. Histological analysis showed a decrease in the number and size of the intestinal microvillae and a light edema of serosa. CONCLUSION: Gastroschisis had a direct correlation with IUGR and the time of exposure of the fetuses to AF had no influence on body weight in gastroschisis fetuses but did interfere with intestinal length. PMID- 16260883 TI - Pseudotrisomy 13: clinical findings and genetic implications. AB - The combination of holoprosencephaly, postaxial polydactyly, and normal karyotype has been termed pseudotrisomy 13 syndrome. Here, we report the prenatal diagnosis of pseudotrisomy 13 in three siblings suggesting autosomal recessive inheritance of this syndrome. Clinical overlap with hydrolethalus syndrome, Smith-Lemli-Opitz syndrome, Meckel syndrome, and Pallister-Hall syndrome is discussed. PMID- 16260882 TI - Isolated fetal hyperechogenic bowel associated with intra-uterine parvovirus B19 infection. AB - We report a case of fetal hyperechogenic bowel diagnosed at midgestation that was associated with fetal parvovirus B19 infection. Isolated hyperechogenic bowel was detected at 25 weeks. Cystic fibrosis, chromosomal abnormalities and cytomegalovirus infection were excluded, whereas polymerase chain reaction DNA for parvovirus B 19 was found positive on amniotic fluid. The hyperechogenic bowel decreased with complete resolution by 32 weeks of gestation. No other signs of fetal B19 infection were detected prenatally and the baby had normal postnatal outcome. This case provides additional arguments in favor of a possible intestinal tropism of parvovirus B19 during fetal life. Fetal B19 infection should be systematically incorporated in the prenatal evaluation of isolated fetal hyperechogenic bowel. PMID- 16260884 TI - Prenatal diagnosis of a fetal chest wall cystic lymphangioma using ultrasonography and MRI: a case report with literature review. AB - A case of fetal chest wall cystic lymphangioma diagnosed prenatally at 28 weeks' gestation is reported. Ultrasound examination showed a multilocular, large cystic mass (10 x 22 x 29 mm) on the left side of the fetus in the area of the lower chest and upper abdomen, without color flow imaging. Magnetic resonance imaging was used to evaluate the extent and the tissue characteristics of the lesion. Two months after birth the lymphangioma was surgically removed, following an infection and a rapid increase in size. The case is discussed, and a short review of the literature is reported. PMID- 16260885 TI - Endothelial nitric oxide synthase polymorphism is not associated with placental abruption in Finnish women. AB - OBJECTIVE: To study whether genetic variability in the gene encoding endothelial nitric oxide synthase (eNOS) affects individual susceptibility to the development of placental abruption during pregnancy. METHODS: One hundred and sixteen pregnant women with placental abruption and 113 healthy controls were genotyped for Glu298Asp polymorphism in the eNOS gene. Chi-square analysis was used to assess the differences in genotype and allele frequencies between the two groups. RESULTS: A statistically similar allelic distribution of eNOS Glu298Asp polymorphism was observed in the two groups, with the frequency of the variant G allele being 66.8% in the abruption group and 68.1% in the control group (OR 0.94, 95% CI: 0.64-1.39; p = 0.76). The genotype distribution of the eNOS polymorphism was also found to be statistically similar (p = 0.72). CONCLUSIONS: The observed genotype data in subjects from eastern Finland suggest that the Glu298Asp polymorphism of the eNOS gene does not contribute to placental abruption in this population. PMID- 16260886 TI - Congenital myotonic dystrophy. AB - We describe a case of severe congenital myotonic dystrophy (CDM). A 38-year-old primigravida, who was known to suffer from mild myotonic dystrophy (DM), conceived spontaneously and booked for confinement at 11 weeks in our unit. The couple had been fully counseled about the risks of transmission of this condition to their offspring before embarking on this pregnancy. Despite being fully aware of the risks, they declined prenatal diagnosis. The pregnancy was monitored by serial ultrasound scans. The diagnosis of CDM was suspected by ultrasound markers of borderline ventriculomegaly, polyhydramnios, and reduced fetal movements. The pregnancy ended prematurely at 33 weeks in an emergency caesarean section because of severe fetal compromise. The neonate died almost immediately after birth. The genetic analysis of cord blood confirmed severe DM. This case highlights the importance of ultrasound markers for the diagnosis of CDM in the absence of definitive prenatal diagnosis. PMID- 16260887 TI - Periconceptional clinics: a medical health care infrastructure of new genetics. AB - The necessary paradigm shift of medical prevention and health promotion from general prevention to specific genetic-oriented prevention require two crucial points: the selection of the optimal time for the primary prevention of birth defects and predictive genetic testing and the establishment of the appropriate healthcare infrastructure. The optimal time for the primary prevention of birth defects (e.g. neural-tube defects) and predictive genetic testing is the preconceptional period, i.e. the preparatory time for the planned conception, particularly before the first pregnancy, and the most acceptable time for the concrete diagnosis of the expected serious genetic diseases is the early postconceptional period. Theoretically, preimplantation genetic diagnosis is the optimal option for offspring of couples at high risk including autosomal and X linked recessive, dominant disorders and chromosomal aberrations. The pre- and early postconceptional health can be addressed jointly in a new type of the health care infrastructure entitled periconceptional clinic. Periconceptional clinics seem to be appropriate for the starting point for the primary prevention of common complex diseases as well. PMID- 16260888 TI - Antenatal detection of arterio-arterial anastomoses by Doppler placental assessment in monochorionic twin pregnancies. AB - OBJECTIVES: To evaluate the reproducibility of Doppler antenatal detection of arterio-arterial anastomoses (AAA) in monochorionic (MC) twin pregnancies. METHODS: Between October 2002 and February 2004, 21 MC diamniotic twin pregnancies and one dichorionic triamniotic triplet seen at the Twin Clinic at the University of Brescia were recruited. After routine ultrasonographic assessment, AAA were searched using Color or Power and spectral Doppler. The presence of AAA was confirmed postnatally by placental injection studies. RESULTS: Data of 19 patients were available for the analysis. AAA were detected in 12 cases (63%) antenatally and in 16 (84.2%) at injection study. Sensitivity and specificity of Doppler for detecting AAA were 75 and 100%, respectively. Detection rates increased at advanced gestations and with anterior/fundal placentae. The incidence of twin-twin transfusion syndrome was higher in the group with no AAA detected in vivo compared to the group with AAA found with Doppler (28.5 vs. 16.6%), but the difference was not statistically significant (p = 0.5). CONCLUSION: This study confirmed the feasibility of AAA Doppler detection in vivo in MC pregnancies. PMID- 16260889 TI - Fetal MRI of a severe Dandy-Walker malformation with an enlarged posterior fossa cyst causing severe hydrocephalus. AB - We present a case of severe Dandy-Walker malformation with enlarged posterior fossa cyst extruding through the incisura of the tentorium and causing severe hydrocephalus. A posterior fossa malformation was suspected by ultrasonography and was further evaluated by fetal magnetic resonance imaging (MRI). Fetal MRI demonstrated that there were no associated anomalies and the decision was made to continue with labor instead of terminating the pregnancy. The precise diagnosis and absence of associated anomalies revealed by MRI aided in parent counseling and obstetrical management. PMID- 16260890 TI - Rare case of exomphalos complicated with umbilical cord teratoma in a fetus with trisomy 13. AB - An exomphalos containing unusual solid and cystic mass was diagnosed during a routine ultrasound examination in the 17th week of gestation. Further investigations were planned but the pregnancy was terminated. The fetopathological examination revealed an umbilical cord teratoma. Although this entity is very rare it should be emphasized as a possible differential diagnosis when cystic lesion of the cord is detected. Large teratomas associated with abdominal wall defect may have poor fetal outcome and can be associated with structural and chromosomal abnormalities. In our case trisomy 13 was diagnosed. PMID- 16260891 TI - Effects of ibuprofen and indomethacin on urinary antidiuretic hormone excretion in preterm infants treated for patent ductus arteriosus. AB - OBJECTIVE: To compare the effects of intravenous ibuprofen and indomethacin for treatment of patent ductus arteriosus (PDA) on urinary antidiuretic hormone (ADH) excretion, as a cause of oliguria. STUDY DESIGN: Forty-four respiratory distress syndrome prematures (or= 350 cells/microl) and HIV RNA levels >or= 5 log10 copies/ml (P = 0.0008). Those with AIDS at the time of starting HAART had a less rapid relative decline in incidence (P = 0.0007). CONCLUSIONS: Our results suggest that some groups may experience less rapid relative declines in AIDS incidence than others over the first year on HAART. PMID- 16260905 TI - Dose-dependent influence of didanosine on immune recovery in HIV-infected patients treated with tenofovir. AB - BACKGROUND: Antiretroviral therapy (ART) containing tenofovir disoproxil fumarate (TDF) and didanosine (ddI) has been associated with poor immune recovery despite virologic success. This effect might be related to ddI toxicity since ddI exposure is substantially increased by TDF. OBJECTIVE: To analyze whether immune recovery during ART with TDF and ddI is ddI-dose dependent. DESIGN AND METHODS: A retrospective longitudinal analysis of immune recovery measured by the CD4 T-cell slope in 614 patients treated with ART containing TDF with or without ddI. Patients were stratified according to the tertiles of their weight-adjusted ddI dose: low dose (< 3.3 mg/kg), intermediate dose (3.3-4.1 mg/kg) and high dose (> 4.1 mg/kg). Cofactors modifying the degree of immune recovery after starting TDF containing ART were identified by univariable and multivariable linear regression analyses. RESULTS: CD4 T-cell slopes were comparable between patients treated with TDF and a weight-adjusted ddI-dose of < 4.1 mg/kg per day (n = 143) versus TDF-without-ddI (n = 393). In the multivariable model the slopes differed by -13 CD4 T cells/mul per year [95% confidence interval (CI), -42 to 17; P = 0.40]. In contrast, patients treated with TDF and a higher ddI dose (> 4.1 mg/kg per day, n = 78) experienced a significantly impaired immune recovery (-47 CD4 T cells/microl per year; 95% CI, -82 to -12; P = 0.009). The virologic response was comparable between the different treatment groups. CONCLUSIONS: Immune recovery is impaired, when high doses of ddI (> 4.1 mg/kg) are given in combination with TDF. If the dose of ddI is adjusted to less than 4.1 mg/kg per day, immune recovery is similar to other TDF-containing ART regimen. PMID- 16260907 TI - Spontaneous control of viral load and CD4 cell count progression among HIV-1 seroconverters. AB - OBJECTIVES: To identify factors associated with sustained undetectable viraemia after HIV-1 seroconversion in treatment-naive patients, and to describe concomitant CD4 cell count progression. METHODS: Seroconverters enrolled in CASCADE were assumed to control viraemia if at least two consecutive viral load measurements were < 400/500 copies/ml without treatment. Factors associated with undetectable viraemia were identified through a logistic regression. A joint model was used to describe simultaneously the CD4 cell count progression during and after that period and to identify factors associated with sustained undetectable viraemia. RESULTS: Of 2176 seroconverters, 145 (6.7%) spontaneously controlled viraemia. Women were more likely than men to achieve undetectable viraemia [adjusted odds ratio (OR), 2.12; 95% confidence interval (CI), 1.49 3.12] unlike patients who reported a symptomatic primary infection (adjusted OR, 0.58; 95% CI, 0.36-0.94). AIDS and death rates were significantly lower in patients achieving undetectable viraemia than in the others. The median period of undetectable viraemia was 11.2 months; on average, CD4 cell counts remained stable during that period, and decreased with a mean rate of 5 cells/microl per month thereafter. High CD4 cell count at the beginning of undetectable viraemia and non-symptomatic primary infection favoured the preservation of undetectable viraemia. CONCLUSION: A small proportion of seroconverters appeared to be able to control HIV viraemia spontaneously, mostly those without seroconversion illness and within a few years following seroconversion; this is associated with the benefits of slower CD4 cell count decline and improved long-term prognosis. Such persons should be targeted for in depth investigation. PMID- 16260909 TI - Effect of HIV on work-related injury rates in South African gold miners. AB - OBJECTIVE: Work-related injuries have severe, well-documented economic and social impacts, and injury is a leading cause of death in working adults. As adults of working age are one of the groups most affected by the HIV epidemic, the interaction between work-related injuries and HIV is important. The objective was to calculate the effect of HIV on the rate and severity of work-related injuries by duration of infection. DESIGN AND METHODS: A large, retrospective seroincident cohort of South African gold miners was studied. Data routinely collected by the mines, and assurance company injury data were analysed. HIV-positive and negative miners were compared, allowing the calculation of injury rates and rate ratios. Severity of injuries was measured by the number of days away from work, percentage of permanent disability, and fatalities. RESULTS: Results were available for 1661 HIV-positive and 6166 HIV-negative miners over 10 years. HIV infection increased the rate of work-related injuries overall (adjusted rate ratio, 1.3; 95% confidence interval, 1.1-1.4), but had less effect on severe injuries. Injury rates in HIV-positive men prior to the first positive test were similar to those in HIV-negative men. The injury rate rose soon after the first HIV positive test. After seroconversion there was only weak evidence of an increase in injury rates by duration of infection. CONCLUSION: This is the first study to demonstrate an increase in injury rates in HIV-positive individuals. The increase may reflect direct effects of HIV infection as well as behaviour change once HIV is diagnosed. PMID- 16260910 TI - Using economic threshold analysis to determine the intensity of HIV prevention services for HIV-seropositive persons. AB - OBJECTIVE: Studies of the effectiveness of HIV prevention interventions for persons living with HIV have examined interventions with very wide variation in intensity (from single session interventions to those offered twice-weekly for 6 months); this raises questions about cost-effectiveness. Herein the question is asked: at varied (but specified) levels of HIV transmission rates and intervention effectiveness, how much money can be spent per HIV-seropositive client on HIV prevention services and still be considered cost-saving to society (in the United States)? DESIGN AND METHODS: Standard methods of economic evaluation (threshold analysis, in particular) were used. Per-client HIV transmission rates and intervention effectiveness were specified and then allowed to vary widely. The threshold for intervention cost (as well as number of sessions per client) that could be justified on the basis of societal cost savings was then calculated. RESULTS: If HIV transmissions are permanently prevented and lifetime medical costs are avoided in an HIV-seronegative partner, then monthly counseling sessions, even after high quality pre- and post-test counseling, and even at moderate levels of effectiveness, may be cost-saving to society (and at higher transmission rates and effectiveness, dozens of sessions per client could be cost-saving). If these interventions delay (but do not permanently prevent) HIV infection among HIV-seronegative partners, then the cost and number of session thresholds are substantially lower. CONCLUSIONS: Even with considerable uncertainty in input parameters, an economic evaluation threshold analysis framework can yield useful insights for guiding the selection of intensity of HIV prevention services for persons living with HIV. PMID- 16260908 TI - Patterns of the hazard of death after AIDS through the evolution of antiretroviral therapy: 1984-2004. AB - OBJECTIVE: To characterize changing survival patterns after development of clinical AIDS from 1984 to 2004, when different antiretroviral therapies were being introduced. DESIGN: Cohort of homosexual men since 1984 and cohort of women since 1994. METHODS: A total of 1504 men and 461 women were followed for all cause mortality after an incident AIDS diagnosis. Relative hazards of death and relative times to death were determined in five therapy eras: no/monotherapy (July 1984-December 1989), monotherapy/combination therapy (January 1990-December 1994), HAART introduction (January 1995-June 1998), short-term stable HAART use (July 1998-June 2001), and moderate-term stable HAART use (July 2001-December 2003). RESULTS: A total of 1057 (54%) study participants died. The time at which 25% of individuals died after an AIDS diagnosis increased significantly from 0.56 years [95% confidence interval (CI), 0.50-0.64] in the no/monotherapy era to 0.74 (95% CI, 0.67-0.82), 1.78 (95% CI, 1.29-2.44), 4.22 (95% CI, 2.94-6.05) and 5.08 years (95% CI, 2.39-10.79) in the four subsequent therapy eras, respectively. Inferences on the beneficial effects of HAART were confirmed after adjustment by age, sex, type of AIDS diagnosis and CD4 cell count at diagnosis. The pattern of the hazard of death after AIDS changed from increasing in the pre-HAART era to being lower and non-increasing in the eras of HAART. CONCLUSIONS: The sustained beneficial effect of HAART, even in individuals with clinical AIDS and extensive treatment histories, attenuates concerns about emergence of resistance but augurs that a substantial number of HIV-infected individuals may require care for very long periods. PMID- 16260911 TI - Renal tubular toxicity associated with tenofovir assessed using urine-beta 2 microglobulin, percentage of tubular reabsorption of phosphate and alkaline phosphatase levels. AB - Despite its wide use, the renal tubular toxicity of tenofovir has not been fully evaluated. Twelve weeks after initiating a tenofovir-containing HAART regimen, a high urine-beta 2 microglobulin level was observed in 12 out of 17 patients, the percentage of tubular reabsorption of phosphate decreased from 96.0 to 91.1% and alkaline phosphatase increased from 294 to 365 U/l, whereas serum creatinine and phosphorus remained largely unchanged. Patients with the above findings should be monitored carefully for renal tubular toxicity. PMID- 16260912 TI - HIV testing among female sex workers in Andhra Pradesh, India. AB - Of 6648 female sex workers (FSW) in 13 districts of Andhra Pradesh state in India, only 7.9% reported having undergone HIV testing, and three-quarters of the rest were unwilling to undergo HIV testing in the future. The risk of HIV infection as a result of the non-use of condoms was higher among FSW who reported not having been tested and were also unwilling to get tested, and they also held significantly more negative beliefs about HIV/AIDS. PMID- 16260913 TI - Risk of selecting K65R in antiretroviral-naive HIV-infected individuals with chronic hepatitis B treated with adefovir. AB - Seven antiretroviral-naive HIV-infected individuals with chronic hepatitis B treated with adefovir for longer than 6 months were assessed. Using bulk population sequencing and a sensitive limiting dilution analysis, the selection of K65R or other resistance mutations did not occur in HIV, suggesting that adefovir can be confidently used as hepatitis B virus (HBV) therapy in HIV/HBV-co infected patients who do not require antiretroviral therapy. PMID- 16260914 TI - Performance of a T-cell-based diagnostic test for tuberculosis infection in HIV infected individuals is independent of CD4 cell count. AB - The performance characteristics of the enzyme-linked immunospot assay (ELISPOT) assay (T-SPOT TB) for the diagnosis of latent tuberculosis infection in HIV infected individuals are unknown. Given that ELISPOT enumerates Mycobacterium tuberculosis antigen-specific IFN-gamma-secreting T cells, HIV-associated immunosuppression might adversely affect test performance. However, we found that 28 out of 29 HIV-positive individuals (97%) gave evaluable test results, and performance was independent of the CD4 T-cell count. ELISPOT test performance appears to be independent of HIV-associated immunosuppression. PMID- 16260915 TI - Potential hazard of pharmacokinetic interactions between lopinavir-ritonavir protease inhibitors and irinotecan. PMID- 16260916 TI - Response to Foss et al., 'Care should be taken when promoting microbicide use among sex workers who are able to use condoms consistently'. PMID- 16260917 TI - Zidovudine-induced pure red cell aplasia presenting after 4 years of therapy. PMID- 16260918 TI - Pulmonary Mycobacterium celatum immune restoration disease: immunopathology and response to corticosteroid therapy. PMID- 16260919 TI - Response to 'Does immune reconstitution promote active tuberculosis in patients receiving highly active antiretroviral therapy?' AIDS, 22 July 2005. PMID- 16260920 TI - Cryptococcocal immune reconstitution disease: a major cause of early mortality in a South African antiretroviral programme. PMID- 16260921 TI - Spinal epidural lipomatosis: a manifestation of HAART-associated lipodystrophy. PMID- 16260922 TI - Atazanavir/ritonavir versus lopinavir/ritonavir: equivalent or different efficacy profiles? PMID- 16260923 TI - Clinical factors associated with employment among people with severe mental illness: findings from the employment intervention demonstration program. AB - Research has shown that supported employment programs are effective in helping psychiatric outpatients achieve vocational outcomes, yet not all program participants are able to realize their employment goals. This study used 24 months of longitudinal data from a multisite study of supported employment interventions to examine the relationship of patient clinical factors to employment outcomes. Multivariate random regression analysis indicated that, even when controlling for an extensive series of demographic, study condition (experimental versus control), and work history covariates, clinical factors were associated with individuals' ability to achieve competitive jobs and to work 40 or more hours per month. Poor self-rated functioning, negative psychiatric symptoms, and recent hospitalizations were most consistently associated with failure to achieve these employment outcomes. These findings suggest ways that providers can tailor supported employment programs to achieve success with a diverse array of clinical subpopulations. PMID- 16260924 TI - Personality differences in schizophrenia are related to performance on neuropsychological tasks. AB - Altered cognition and personality appear to emerge in tandem and adversely affect outcome in schizophrenia, yet little research has been done to determine whether these are related or independent domains. In this study, the relationship between the Big Five personality traits--neuroticism, extraversion, openness, agreeableness, conscientiousness--and cognitive and motor performance in outpatients with chronic, clinically stable schizophrenia (N = 30) and age matched healthy comparison subjects (N = 45) was examined. Subjects completed tests of attention, executive and motor functions, and the NEO-Five Factor Personality Inventory. Patients scored significantly higher on neuroticism and lower on extraversion and agreeableness, but after variance due to neuropsychological performance was statistically removed from NEO scale scores, personality dimensions and profiles no longer differed between groups. Neuropsychological performance and demographic variables, but not diagnosis, uniquely accounted for statistically significant amounts of personality variance, and neuropsychological task performance was correlated with personality dimensions in both patients and comparison subjects. These cross-sectional data provide preliminary evidence that personality dysfunction in schizophrenia may be mediated by disease-related changes in cognitive operations, or the neural processes underlying them. Longitudinal studies utilizing more comprehensive measures of neurocognitive performance are needed to define further the relationship between neuropsychological function and personality in schizophrenia. PMID- 16260925 TI - Replication of dissociation-psychosis link in New Zealand students and inmates. AB - To assess the relationship between dissociative and psychotic experiences, New Zealand university students (N = 119) and prison inmates (N = 42) were administered the Dissociative Experiences Scale (DES) and Symptom Checklist-90 Revised (SCL-90-R). Strong correlations were found between DES scores and the psychoticism and paranoid ideation subscales of the SCL-90-R (students: r = .520, .517, respectively; inmates: r = .637, .649, respectively). While other correlations were also significant (but smaller), these results are consistent with previous studies that have used a range of measures of psychosis or schizotypy with a variety of clinical and nonclinical populations. Such consistent findings in the face of methodological diversity offer strong support for the validity of a link between the concepts of dissociation and psychosis. While this relationship has previously been interpreted indirectly, as dissociative experiences predisposing to psychotic symptoms, we suggest a direct route: that dissociative experiences of various forms may underlie some (or even all) psychotic symptoms. PMID- 16260926 TI - Proposed diagnostic criteria of Internet addiction for adolescents. AB - The aims of the present study were to develop diagnostic criteria of Internet addiction for adolescents and to examine the discriminating potential and validity of diagnostic criteria by an empirical community study among adolescent populations. We developed 13 candidate diagnostic criteria for characteristic symptoms of Internet addiction in adolescents. A total of 468 adolescents completed the Chen Internet Addiction Scale (CIAS) and were systematically assessed for Internet-using behaviors by seven psychiatrists using the diagnostic interview schedule. The sensitivity, specificity, and diagnostic accuracy of the 13 candidate diagnostic criteria were analyzed with references to the interviewers' global clinical impressions and CIAS results. The cutoff point of the diagnostic criteria to differentiate the Internet-addicted subjects with nonaddicted ones was then determined by the best diagnostic accuracy and the receiver operating characteristic curve. This study selected nine of the 13 candidate diagnostic criteria to construct the diagnostic criteria of Internet addiction for adolescents, which were composed of three main criteria: characteristic symptoms of Internet addiction, functional impairment secondary to Internet use, and exclusive criteria. The diagnostic criteria had high diagnostic accuracy, specificity, negative predictive value, accepted sensitivity, and accepted positive predictive rate. The validity of the diagnostic criteria proposed in this study was further confirmed by comparing the demographic and Internet-using characteristics between those with and without Internet addiction. The diagnostic criteria for Internet addiction can provide health care professionals with a means to communicate and make comparisons of clinical cases. PMID- 16260927 TI - Rosenhan revisited: the scientific credibility of Lauren Slater's pseudopatient diagnosis study. AB - In a recent and widely publicized book, psychologist Lauren Slater reported an attempt to test David Rosenhan's hypothesis that psychiatric diagnoses are influenced primarily by situational context rather than by patients' signs and symptoms. Slater presented herself to nine psychiatric emergency rooms with the lone complaint of an isolated auditory hallucination (hearing the word "thud"). In almost all cases, she reported receiving the diagnosis of psychotic depression and prescriptions for antidepressants and antipsychotics. Slater concluded that psychiatric diagnoses are largely arbitrary and driven by a "zeal to prescribe." Our goal was to examine the scientific credibility of Slater's findings using a vignette methodology. We presented a sample of emergency room psychiatrists (N = 74) with a detailed case vignette derived from the clinical description in Slater's book, and asked them a series of questions regarding diagnosis and treatment recommendations. In sharp contrast to what Slater reported, we found that only three psychiatrists offered a diagnosis of psychotic depression. Moreover, only one third recommended medication. Our study raises questions regarding Slater's results and conclusions, and provides scant support for the claim that psychiatric diagnoses are mostly products of fashion or fad, as claimed by Slater. PMID- 16260929 TI - Reply to Spitzer and colleagues. PMID- 16260928 TI - Pseudopatient or pseudoscience: a reviewer's perspective. PMID- 16260930 TI - A response to a nonresponse to criticisms of a nonstudy: one humorous and one serious rejoinder to Slater. PMID- 16260931 TI - Personality disorders and the trimensional personality questionnaire factors in major depressive disorder. AB - We assessed the potential relationship between personality disorder (PD) clusters, as assessed by the SCID-II, and temperamental traits assessed by the Tridimensional Personality Questionnaire (TPQ) among a well-characterized, unmedicated cohort of outpatients with major depressive disorder (MDD). The TPQ and SCID-II were administered to 263 depressed outpatients (mean age = 39.9 +/- 10.5 years; women = 139, 53%; initial 17-item Hamilton Rating Scale for Depression = 19.6 +/- 3.4) who currently met criteria for MDD and who were enrolled in an 8-week treatment trial. The multiple linear regression method was used to evaluate the relationship between TPQ factors and personality disorder clusters, controlling for age, gender, and initial 17-item Hamilton Rating Scale for Depression score as necessary. Among outpatients with MDD, meeting criteria for a Cluster A PD diagnosis was related to high harm avoidance (HA) scores, as well as low reward dependence and novelty seeking (NS) scores. Additionally, high HA scores were associated with meeting criteria for a Cluster C PD diagnosis, while high NS scores were associated with meeting criteria for a Cluster B PD diagnosis. Certain temperament traits, especially HA and NS, appear to be associated with specific patterns of personality clusters among depressed patients. PMID- 16260932 TI - Social cognition and symptom dimensions. AB - This study used the Picture Arrangement subtest of the Wechsler Adult Intelligence Scale to assess social cognitive functioning of psychotic patients diagnosed with bipolar, schizophrenia, or schizoaffective disorder. All participants were rated on positive and negative symptom rating scales, from which three symptom dimensions were obtained. Symptom dimensions were not significantly related to ratings of symptom severity or mental status examination scores. Disorganized symptoms were correlated with a wide range of impairments on nearly all subscale measures of social cognition. Negative symptoms were correlated with lower ratings of capacity for emotional investment, complexity of representations, and integration of episodes. Psychoticism was associated with negative affect tone. Diagnostic categories were not related to social cognitive impairments independent of estimates of premorbid level of cognitive function. Evidence of syndromal differences suggests that heterogeneity of variance in studies of functioning of schizophrenic patients can be reduced by inclusion of symptom dimensions. PMID- 16260933 TI - A study of the face validity of the 40 item version of the Defense Style Questionnaire (DSQ-40). AB - There are few studies examining the face validity of the 40-item version of the Defense Style Questionnaire (DSQ-40). Moreover, the existing studies have provided conflicting results. The present study provides an in-depth examination of the face validity of the DSQ-40. Eight clinicians independently attributed each item of the DSQ-40 to a defense mechanism. The defense mechanisms listed in the DSM-IV Defensive Functioning Scale and their definitions were provided as a guide, along with the definition of those defense mechanisms investigated by the DSQ that are not included. It was further specified that the raters could attribute the items to defense mechanisms other than those listed or coping mechanisms. Twelve items out of 40 (30%) were attributed to the defense mechanisms they were supposed to investigate by fewer than four out of the eight raters. This result suggests that a substantial part of the DSQ-40 is lacking in face validity. PMID- 16260934 TI - Unaccompanied adolescents seeking asylum: poorer mental health under a restrictive reception. AB - We assessed the effects of a stringent reception policy on the mental health of unaccompanied adolescent asylum seekers by comparing the mental health of adolescents in a restricted campus reception setting and in a setting offering more autonomy (numbers [response rates]: 69 [93%] and 53 [69%], respectively; mean age, 16 years). Unaccompanied adolescent asylum seekers in a restricted reception setting reported more emotional problems on the Hopkins Symptom Checklist than their counterparts in the more autonomy group (mean scores [SD]: restricted, 59.3 [13.1]; other, 53.4 [10.5]; p = 0.033, F test). Main effects concerned a rise in anxiety. Girls showed larger differences than boys. A restrictive reception may therefore affect the mental health of minor asylum seekers. Health care workers and policy makers should be aware of this adverse effect. PMID- 16260935 TI - Narcissistic vulnerability and the development of PTSD: a prospective study. AB - This study empirically examined the role of narcissistic traits and narcissistic vulnerability in the development of post-traumatic stress disorder (PTSD). One hundred forty-four survivors of a traumatic event were assessed 1 week, 1 month, and 4 months following the event. In the first-week assessment, patients were administered the Narcissistic Vulnerability Scale and self-reported rating scale to assess event severity and symptoms ensuing from the impact of the traumatic event: depression, intrusions, avoidance, and arousal. In the follow-up assessments, subjects were interviewed on the Clinician-Administered PTSD Scale and were readministered the self-rating symptoms scale. Survivors who developed acute (1 month) and chronic (4 months) PTSD had significantly higher levels of narcissistic vulnerability in the first-week assessment. Narcissistic Vulnerability Scale scores predicted PTSD status with sensitivity of 81.6% and 85.1% and specificity of 40.4% and 38.6% at the 1-month and 4-month assessments, respectively. Narcissistic vulnerabilities contribute to the occurrence of PTSD. PMID- 16260936 TI - Changing the attitudes of Hong Kong medical students toward people with mental illness. AB - This study examines the personal and attitudinal variables that are associated with helping behavior in a hypothetical general practice setting. We explored the effect of an antistigma seminar during a psychiatric clerkship on medical students' attitudes toward the mentally ill. We randomly assigned three rotations of students (81 students) to receive the seminar and three rotations (85 students) as controls. The students expressed views about patients with schizophrenia or depressive disorder portrayed in video vignettes. How dangerous the students perceived target individuals to be was the major determinant of helping behavior. The students' gender, religious affiliation, affective reaction, skill assessment, and controllability attribution were less consistent in predicting behavior. Exposure to the seminar and clerkship experience significantly improved attitudes, but attributes of responsibility and readiness to provide medical care for psychiatric patients were the most resistant to change. We identified certain issues that should be highlighted in future antistigma programs. PMID- 16260937 TI - Adolescent attitudes toward serious mental illness. AB - While there is a growing literature on mental illness stigma and strategies for reducing stigma among adults, less is known about how children and adolescents view persons with mental illness. In this paper, we describe the Attitudes Toward Serious Mental Illness Scale-Adolescent Version (ATSMI-AV) and our initial examinations of its factor structure and variations among subgroups of adolescents. Findings suggest that strategies aimed at reducing stigma in this age group would be wise to specifically target categorical thinking about mental health and mental illness and perceptions of persons with mental illness being violent and out of control. PMID- 16260939 TI - The context & clinical evidence for common nursing practices during labor. AB - The purpose of this article is to review the context and current evidence for common nursing care practices during labor and birth. Although many nursing interventions during labor and birth are based on physician orders, there are a number of care processes that are mainly within the realm of nursing practice. In many cases, particularly in community hospitals, routine physician orders for intrapartum care provide wide latitude for nurses in how they ultimately carry out those orders. An important consideration of common nursing practices during labor is the context or practice model in which those practices occur. Nursing practice is not the same in all clinical environments. Intrapartum nursing practice consists of an assortment of different roles depending on the circumstances, hospital setting, and context in which it takes place. A variety of intrapartum nursing practice models have evolved as a result and in response to the range of sizes, locations, and provider practice styles found in hospitals providing obstetric services. A summary of intrapartum nursing models is presented. The evidence is reviewed for the three most common clinical practices for which nurses have primary responsibility in most settings and that comprise the majority of their time in caring for women during labor: (1) maternal-fetal assessment, (2) management of oxytocin infusions, and (3) second-stage care. Evidence exists for these nursing interventions that can be used to promote maternal-fetal well-being, minimize risk, and enhance patient safety. PMID- 16260941 TI - Effectiveness of adolescent smoking prevention strategies. AB - In the past 30 years, nursing knowledge of adolescent healthcare has grown tremendously, and the added knowledge can help us improve the care we provide to adolescents using evidence-based interventions for practice. While substantial progress has been made in reducing the numbers of adolescents who smoke, tobacco use by adolescents continues to be one of the largest public health problems in the United States. Current estimates are that about one in four adolescents smokes cigarettes on a regular basis. Researchers have tested community-based, school-based, and family-based interventions to prevent tobacco use among teens. This article revisits these interventions, summarizes those that have demonstrated effectiveness in preventing tobacco use among adolescents, and suggests how the interventions can be incorporated into nursing practice with teens. Recommendations for further research to establish evidence-based practice also are offered. PMID- 16260942 TI - Thirty years of maternal-child health policies in the community. AB - This article reviews outcomes, indicators, and challenges for building evidence based practice in community maternal-child health (MCH), and includes promising new design and analytical strategies. In addition, 10 topic areas are listed, which are the foundation of community MCH evidence: (1) evidence of health behavior on mortality/morbidity; (2) theoretical underpinnings of public policy interventions; (3) evidence of growing health disparities; (4) the potential of exploding information technologies; (5) data on aging, maternity, employment, and lactation; (6) data on the changing face of HIV/AIDS; (7) data on the changing way we give birth; (8) drug safety registries; (9) antibiotic-resistant organisms; and (10) environmental pollutants and health. In addition, evidence of indirect and global influences on community MCH is reviewed and the principles of lifestyle change and health promotion are emphasized. PMID- 16260943 TI - What has the literature taught us about culturally competent care of women and children. AB - This article describes what is currently in the literature about culturally competent care for women and children. With the population of the United States growing increasingly diverse, there is a developing need for cultural competency among nurses and throughout healthcare organizations. Cultural competence includes both culture-specific and culture-generic knowledge, attitudes, and skills. While databased literature on cultural competency still requires further development, we do have evidence of positive outcomes of culturally competent care. The end result of the provision of culturally competent care by culturally competent nurses and healthcare organizations can be significant improvements in the health and well-being of women and children. PMID- 16260944 TI - Evidence for healing interventions with perinatal bereavement. AB - The purpose of this article is to explore the concept of perinatal grief and evidence-based healing interventions for it. The loss of a pregnancy or death of an infant causes profound grief, yet society has long minimized or ignored this grief, which is among the most painful of bereavement experiences. Throughout the last century, research on grief and the special needs of bereaved parents has changed the context of professional intervention from protective to supportive. The central focus of bereavement interventions is to assist families in healing by helping them make meaning of their losses. The use of symbols, spirituality, and rituals has been shown to help bring meaning. Research has shown that memories are key to healing, and that gender, age, and relationships bring different grief expressions and experiences. While children's understanding of loss and grief differs with developmental age, they should also be given the opportunity to participate in grief rituals and practices. Professionals who care for bereaved parents have a unique opportunity to offer support by validating their grief, facilitating rituals, providing mementos, and letting the bereaved tell their stories. While no intervention can bring back their beloved children, appropriate intervention can promote healing. PMID- 16260945 TI - Evidence for best practices in the neonatal period. AB - The purpose of this article is to provide a review of current nursing research that supports best practices during the newborn period. The literature review of peer-reviewed research articles published between January 2000 and October 2004 was conducted via keyword searches using the databases of the Cochrane Library, CINAHL, and MEDLINE. Key words included neonatal nursing, newborn, neonate, premature infant, preterm infant, and low birthweight. Content analysis revealed the following primary categories of studies that provide solid evidence for nursing practice: developmentally focused nursing care, neonatal skin care, feeding, skin-to-skin care, and pain management. Neonatal nurse researchers have made many important contributions to the research literature. Future research should expand the findings to date on the effective use of pain scales, the outcomes of skin-to-skin care and infant massage as standard practice for all neonates, and the effectiveness of nursing interventions to support the developmental sequelae of prematurity. Neonatal nurses should become familiar with and implement those findings from nursing research that strongly support evidence-based nursing practice. PMID- 16260947 TI - Where is the evidence for teaching methods used in nursing education? AB - Twenty-first century maternal-child faculty members are now being encouraged to base their teaching practice on sound evidence. Finding that evidence, however, can be challenging. This article discusses not only the current state of research on teaching in nursing, but also how and where educators can access information about teaching that is based on research. It also describes strategies educators can use to develop an evidence-based approach to teaching. Topics included in this discussion include recommended literature resources in nursing and education, bibliographic database search techniques, guides for appraisal of quantitative and qualitative research, and relevant nursing education and general education professional organizations. In addition, an evidence hierarchy from the field of education is introduced to rank educational studies in nursing. If nurse educators are to apply the evidence-based process used in clinical practice to the practice of nursing education, more research is needed in this field. PMID- 16260948 TI - Back safety for nurses. PMID- 16260962 TI - Chronic fatigue syndrome: exercise performance related to immune dysfunction. AB - PURPOSE: To date, the exact cause of abnormal exercise response in chronic fatigue syndrome (CFS) remains to be revealed, but evidence addressing intracellular immune deregulation in CFS is growing. Therefore, the aim of this cross-sectional study was to examine the interactions between several intracellular immune variables and exercise performance in CFS patients. METHODS: After venous blood sampling, subjects (16 CFS patients) performed a maximal exercise stress test on a bicycle ergometer with continuous monitoring of cardiorespiratory variables. The following immune variables were assessed: the ratio of 37 kDa Ribonuclease (RNase) L to the 83 kDa native RNase L (using a radiolabeled ligand/receptor assay), RNase L enzymatic activity (enzymatic assay), protein kinase R activity assay (comparison Western blot), elastase activity (enzymatic-colorimetric assay), the percent of monocytes, and nitric oxide determination (for monocytes and lymphocytes; flow cytometry, live cell assay). RESULTS: Forward stepwise multiple regression analysis revealed 1) that elastase activity was the only factor related to the reduction in oxygen uptake at a respiratory exchange ratio (RER) of 1.0 (regression model: R = 0.53, F (1,14) = 15.5, P < 0.002; elastase activity P < 0.002); 2) that the protein kinase R activity was the principle factor related to the reduction in workload at RER = 1.0; and 3) that elastase activity was the principle factor related to the reduction in percent of target heart rate achieved. CONCLUSION: These data provide evidence for an association between intracellular immune deregulation and exercise performance in patients with CFS. To establish a causal relationship, further study of these interactions using a prospective longitudinal design is required. PMID- 16260963 TI - Intramuscular hemangioma of the foot: a diagnostic dilemma. AB - Intramuscular hemangioma is a rare but important cause of pain and potentially decreased performance in athletes. These benign tumors occur more often in the lower extremity and usually present during the first three decades of life. Symptomatic intramuscular hemangiomas usually present with pain and swelling, which often worsen with activity. Diagnosis can be difficult, resulting in frustration and prolonged activity limitations for these patients. Magnetic resonance imaging and clinical findings can often suggest the diagnosis. Recommended treatment is usually wide total excision, but there are certain cases in which excision is not possible. In difficult areas such as the hands or feet, a sclerosing agent can be injected into the hemangioma to reduce the size of the hemangioma, decrease pain, and improve function. PMID- 16260964 TI - Injury rates in house league, select, and representative youth ice hockey. AB - PURPOSE: The purpose of this study was to determine injury rates in a youth ice hockey program over two seasons (2002-2004). Injury rates for age groups (4-18 yr) and for different levels of competition were compared. Another purpose was to determine the effect of body checking on injury rates among these youths. METHODS: A prospective injury report form was completed by a volunteer trainer for each injury that caused a loss of player time and resulted in evaluation by a physician. The injury form documented age group, type of injury, length of time that the player missed action due to the injury, location of the injury, and circumstances that led to the injury. Participants included 2632 boys aged 4-18 who played in the 2002-2003 season and 2639 boys who played in the 2003-2004 season. RESULTS: Injuries were four times more likely to occur in games than practices. Boys who played in the most advanced levels of competition are 6.1 times more likely to be injured than boys playing in house leagues. Injury rates during games showed a trend toward increasing with the age of the player. Injury rates spiked the first year that body checking was introduced in two different competition levels. Injury rates also spiked with the onset of adolescence (age 13). CONCLUSION: The study findings suggest that the introduction of body checking at age 9 to competitive youth hockey causes an immediate but relatively short-term increase in injury rates. The period of adjustment that accompanies body checking should be taken into account when determining the age at which body checking is introduced. PMID- 16260965 TI - Effects of muscle strength training and testosterone in frail elderly males. AB - PURPOSE: Determine the independent and combined effects of progressive resistance muscle strength training (PRMST) and testosterone on strength, muscle mass, and function in hypogonadal elderly male recuperative care patients. METHODS: Between 1999 and 2004, 71 subjects (mean age 78.2 +/- 6.4 yr, 86% white) were enrolled. After baseline one-repetition maximum (1RM) strength testing and then randomization to one of four treatment groups (low-resistance (20% of the 1RM) exercises and weekly injections of either 100 mg of testosterone enanthate or placebo or high-intensity PRMST (> or =80% 1RM) and weekly injections), each subject received training and injections for 12 wk. RESULTS: Ten subjects withdrew from the study before its completion. Based on intent-to-treat analyses, strength improved in all groups, but was greater with high-intensity PRMST compared with low-resistance exercise (e.g., leg press, (mean +/- SE), 28 +/- 4 vs 13 +/- 4%, P = 0.009). Although testosterone led to significantly greater increases in midthigh cross-sectional muscle area compared with placebo (7.9 +/- 1.3 vs 2.4 +/- 1.4%, P = 0.005), it produced only a nonsignificant trend toward greater strength gains (e.g., leg press 25 +/- 4 vs 16 +/- 4%, P = 0.144). Change in aggregate functional performance score (the sum of 4 functional performance test scores) did not differ between the four intervention groups nor with high intensity PRMST compared with low-resistance exercise (7 +/- 5 vs 15 +/- 5%, P = 0.263). There was not a significant interaction between exercise and testosterone for any outcome. CONCLUSION: High-intensity PRMST is as safe and well tolerated as a similarly structured low-resistance exercise regimen for very frail elderly patients, but produces greater muscle strength improvements. The addition of testosterone leads to greater muscle size and a trend toward greater strength but did not produce a synergistic interaction with exercise. Neither intervention had a significant effect on functional performance. PMID- 16260966 TI - Spring-levered versus piezo-electric pedometer accuracy in overweight and obese adults. AB - PURPOSE: The purpose of this study was to examine the effects of BMI, waist circumference, and pedometer tilt on the accuracy of a spring-levered pedometer (Yamax Digiwalker SW-200 (SW)) and a piezo-electric pedometer (New Lifestyles NL 2000 (NL)) during treadmill walking and over a 24-h period in overweight and obese adults. METHODS: Forty participants (40 +/- 13.0 yr, 32.6 +/- 4.8 kg.m) walked on a treadmill at various speeds (54, 67, 80, 94, and 107 m.min) for 3-min stages. Simultaneously, an investigator determined actual steps by a hand counter. For all walking trials the SW and NL were positioned on the right and left waistband, respectively. Height, weight, pedometer tilt angle and circumference measures of the hip and waist were also measured. Thirty-six participants wore the pedometers for a 24-h period in the same position as during the treadmill walking trials. RESULTS: : In general, the SW became less accurate with increasing BMI, increasing waist circumference, and greater pedometer tilt, whereas the NL was not affected by these variables. The SW error scores were significantly correlated with the absolute pedometer tilt angle at all walking speeds (P < 0.05), but the NL error scores were not. On average the NL recorded 1030 +/- 1414 (16.5 +/- 22.7%) more steps that the SW during the 24-h trial. CONCLUSION: In overweight and obese individuals, a piezo-electric pedometer (NL) is more accurate than a spring-levered pedometer (SW), especially at slower walking speeds. In addition, it appears that pedometer tilt; more so than waist circumference and BMI, was the most important factor influencing the accuracy of the SW. The NL accuracy was not affected by pedometer tilt, waist circumference, or BMI. PMID- 16260967 TI - Effects of aerobic training on gene expression in skeletal muscle of elderly men. AB - PURPOSE: To analyze the effect of 3 months of training on global gene expression in skeletal muscle of healthy elderly men in order to better characterize the pathways that differentiate the trained from the sedentary state. METHODS: Needle biopsies were obtained from the vastus lateralis of six healthy, sedentary, 68.0 +/- 2.7-yr-old males, before and after 3 months of training at 80% of maximal capacity, that improved peak O2 uptake by 17.8 +/- 4.0% (P = 0.009). RNA extracted from the samples was hybridized to Affymetrix U133A Genechip arrays. RESULTS: Overall, training modified the expression of 397 out of 14,500 genes tested. In the trained state, the expression of genes linked with energy metabolism, protein amino acid dephosphorylation, and heme biosynthesis increased, whereas those linked with ribosome and protein catabolism decreased. CONCLUSION: This set of 397 genes that includes the upregulated energy pathways and the downregulated protein catabolism genes represents the transcriptional response of the skeletal muscle in the trained state. It is highly likely that many of these genes are mediators of the beneficial effects of physical activity on health and fitness. PMID- 16260968 TI - Hyperosmolarity causes BK Ca-dependent vasodilatations in rat skeletal muscle arteries. AB - PURPOSE: The release of different metabolites during skeletal muscle contraction causes a pronounced increase in extracellular osmolarity (hyperosmolarity (HO)). HO has been considered as a possible mediator of the exercise hyperemia. In the present study, we investigated the vasodilatory effect of physiologically relevant increases in the extracellular osmolarity in isolated rat gluteal muscle arterioles. In addition, we analyzed the underlying mechanisms of the HO-induced vasodilatations. METHODS: Rat gluteal arteries were isolated and mounted in an organ bath for isometric tension recording. After precontraction with norepinephrine, 20, 40, or 60 mmol x L(-1) sucrose, mannitol, or urea was added in control conditions, after removal of the endothelium or in the presence of inhibitors. RESULTS: Application of sucrose or mannitol induced large and fast concentration-dependent vasodilatations (up to 46.15% with 60 mmol x L(-1) sucrose). Removal of the vascular endothelium had no effect on this relaxation. Inhibition of the Na+/K+ pumps with ouabain, the Kir IR channels with Ba2+ and the K ATP channels with glibenclamide did not alter the HO-induced relaxations. Incubation with the KCa channel blockers charybdotoxin and apamin significantly inhibited sucrose-induced vasodilatations. In addition, application of the specific BK Ca channel blocker iberiotoxin significantly decreased the HO-induced vasodilatations. CONCLUSION: The present study shows that an increase in the extracellular osmolarity elicits strong, fast, and long-lasting relaxations of rat skeletal muscle arterioles, suggesting an important role both at the onset and during the steady-state phase of an exercise bout. Vascular smooth muscle BK Ca channels seem to play a crucial role in the HO-induced vasorelaxations. PMID- 16260969 TI - Beta1-adrenoceptor mediated origin of the heart rate performance curve deflection. AB - PURPOSE: The deflection of the HR performance curve (HRPC) has been described as an objective marker of submaximal exercise performance. HR response to incremental cycle ergometer exercise is shown to be neither linear nor uniform and a physiological explanation of the deflection phenomenon is lacking. We hypothesized that differences in the beta1-adrenoceptor site are the source of these differences. The aim of the study was to investigate the influence of the highly selective beta1-adrenoceptor (beta1-AR) antagonist bisoprolol (Bi) on the HRPC in young healthy male subjects with different HR response patterns. METHODS: Sixteen subjects were treated in randomized order with Bi or a placebo (Pl) in two separate trials. HR response during incremental cycle ergometer exercise was compared between the two trials. Blood lactate concentration (La) and ventilatory variables were measured throughout both tests. RESULTS: Bi changed the direction of the HRPC more in subjects with a regular, s-shaped response pattern under placebo than those with a nonregular or linear pattern. The influence of Bi on the HR at the second lactate turn point was significantly related (R = 0.78; P < 0.001) to the pattern of the HRPC in Pl conditions. CONCLUSION: We suggest that differences between the subjects with regular s-shaped versus nonregular HRPC may be due to differences at the beta1-AR site. The origin of the HRPC deflection is mediated in part by the beta1-AR sensitivity. PMID- 16260970 TI - Aortic stiffness and aerobic exercise: mechanistic insight from microarray analyses. AB - INTRODUCTION/PURPOSE: Regular aerobic exercise reduces aortic stiffness. However, the mechanisms by which chronic exercise lowers arterial stiffness are not known. To determine the molecular mechanisms of these changes, the alteration of gene expression in the aorta by aerobic exercise training was measured with the microarray technique. METHODS/RESULTS: The differences in expression levels of 3800 genes in the abdominal aorta of sedentary control rats (8 wk old) and exercise-trained rats (8 wk old, treadmill running for 4 wk) were compared by the microarray analysis. Aortic pulse wave velocity (PWV) was lower and systemic arterial compliance was higher (both P < 0.05) in the exercise-trained group than in the control group. Of the 323 genes that displayed differential expression (upregulation of 206 genes and downregulation of 117 genes), a total of 29 genes (24 upregulated and 5 downregulated genes) were identified as potential candidate genes that may be involved in vasodilation and arterial destiffening. Using real time quantitative polymerase chain reaction, we confirmed the results of microarray analysis that prostaglandin EP2 receptor (PGE-EP2R), prostaglandin EP4 receptor (PGE-EP4R), C-type natriuretic peptide (CNP), and endothelial nitric oxide synthase (eNOS) genes were differentially expressed. Furthermore, there were modest correlations between arterial stiffness and levels of these factors. Differential expression of eNOS gene was further verified at protein level by using Western blot analysis. CONCLUSION: These results suggest that exercise training induces the altered expression in several genes including prostaglandin, CNP, and nitric oxide in the aorta and that these molecular changes (particularly eNOS as its protein expression was altered) may contribute, at least in part, to the beneficial effect of exercise training on aortic stiffness. PMID- 16260971 TI - Effect of oral creatine supplementation on urinary methylamine, formaldehyde, and formate. AB - PURPOSE: It has been claimed that oral creatine supplementation might have potential cytotoxic effects on healthy consumers by increasing the production of methylamine and formaldehyde. Despite this allegation, there has been no scientific evidence obtained in humans to sustain or disprove such a detrimental effect of this widely used ergogenic substance. METHODS: Twenty young healthy men ingested 21 g of creatine monohydrate daily for 14 consecutive days. Venous blood samples and 24-h urine were collected before and after the 14th day of supplementation. Creatine and creatinine were analyzed in plasma and urine, and methylamine, formaldehyde, and formate were determined in 24-h urine samples. RESULTS: Oral creatine supplementation increased plasma creatine content 7.2-fold (P < 0.001) and urine output 141-fold (P < 0.001) with no effect on creatinine levels. Twenty-four-hour urine excretion of methylamine and formaldehyde increased, respectively, 9.2-fold (P = 0.001) and 4.5-fold (P = 0.002) after creatine feeding, with no increase in urinary albumin output (9.78 +/- 1.93 mg x 24 h(-1) before, 6.97 +/- 1.15 mg x 24 h(-1) creatine feeding). CONCLUSION: This investigation shows that short-term, high-dose oral creatine supplementation enhances the excretion of potential cytotoxic compounds, but does not have any detrimental effects on kidney permeability. This provides indirect evidence of the absence of microangiopathy in renal glomeruli. PMID- 16260973 TI - Function of the lower esophageal sphincter during and after high-intensity exercise. AB - INTRODUCTION: Gastroesophageal reflux is commonly reported during high-intensity endurance exercise in otherwise healthy asymptomatic individuals. Although the mechanisms underlying this exercise-induced reflux are unknown, it most likely reflects a failure of the primary barrier to reflux, the lower esophageal sphincter (LES). The aim of this study was to determine the influence of exercise with and without ingestion of fluid on the LES barrier pressure in asymptomatic individuals. METHODS: Seven recreational cyclists (five males) performed four 5 min bouts of cycle exercise at 90% VO2max, each separated by 1-3 min. Before, during, and after exercise, measurements were made of esophageal pressure (Pes), LES pressure (Ples), and gastric pressure (Pg). LES barrier pressure (Pb) was defined as the difference between Ples and Pg. Following exercise bouts 2 and 3, subjects ingested 600 and 200 mL of a sports drink, respectively. RESULTS: Pb before exercise was 13.1 +/- 5.2 cm H2O (+/- SD), decreased to 6.5 +/- 4.6 cm H2O during each of the four bouts of high-intensity exercise (P < 0.05), and remained decreased at 7.4 +/- 3.5 cm H2O after exercise (P < 0.05). CONCLUSIONS: High intensity exercise reduces LES Pb during exercise in asymptomatic individuals. The magnitude of this exercise-induced impairment in LES function is unaffected by ingestion of a sports drink. PMID- 16260972 TI - Metabolic responses to exercise after carbohydrate loads in healthy men and women. AB - PURPOSE: This study aimed to investigate gender differences in i) pancreatic insulin secretory (beta-cell sensitivity) and whole body insulin sensitivity responses to an intravenous carbohydrate (CHO) load, and (ii) metabolic responses to exercise after both intravenous and oral CHO loads. METHODS: Seven untrained healthy men and seven age-, body mass-, and VO2max-matched women performed two trials. In one trial they cycled for 60 min at 50% VO2max, starting 60 min after ingestion of a carbohydrate-rich meal (ME trial). In the other trial, subjects were infused with 20% dextrose solution to maintain blood glucose concentration at approximately 8 mmol x L(-1) for 60 min (INF trial), then the infusion rate was maintained constant during the following 60 min while exercising at 50% VO2max. RESULTS: There was no gender effect on beta-cell sensitivity (serum insulin: 161 +/- 37 and 159 +/- 28 pmol x L(-1) for men and women, respectively) and whole body insulin sensitivity (155 +/- 24 and 135 +/- 29 mg x KgFFM(-1) x min(-1) per pmol x L(-1) x 100 for men and women, respectively). This may explain the similarity in glycemic, substrate oxidation and other metabolic responses to exercise after both intravenous and oral CHO loads in men and women. CONCLUSION: These results suggest that moderate exercise performed in the postprandial state presents a similar challenge to the ability of healthy, untrained men and women to perform exercise without a substantial decline in plasma glucose concentration below fasting values. PMID- 16260974 TI - Validity of a single-visit protocol to estimate the maximum lactate steady state. AB - INTRODUCTION: The maximum lactate steady state (MLSS) represents the highest intensity of exercise at which a balance exists between the rate of lactate production and the rate of lactate clearance. The MLSS is an important determinant of endurance exercise performance but its determination, which involves 4-5 laboratory visits, is labor-intensive and time consuming. PURPOSE: To compare the MLSS estimated from a single-visit protocol (MLSSsingle) with the traditional, directly measured MLSS (MLSStrad). METHODS: Following an incremental treadmill test for the determination of VO2peak, eight endurance-trained runners completed: 1) a series of 4-5 constant-speed treadmill runs of up to 30-min duration, on separate days, for determination of the MLSStrad; and 2) a single visit protocol consisting of two constant-speed treadmill runs of 20-min duration at approximately 65 and 90% VO2peak separated by 40 min of rest, for determination of MLSSsingle. Blood lactate concentration ([La]), oxygen uptake (VO2), and heart rate (HR) were measured every 5 min in all treadmill runs. Comparisons between values were made using paired t-tests. RESULTS: The MLSSsingle significantly underestimated the MLSStrad with respect to speed (13.4 +/- 1.2 vs 16.4 +/- 1.6 km x h(-1), P = 0.002), HR (148 +/- 8 vs 170 +/- 10 b x min(-1), P < 0.001), blood [La] (2.3 +/- 1.5 vs 3.9 +/- 1.0 mmol x L(-1); P = 0.01), and % VO2peak utilized (75 +/- 8 vs 90 +/- 2%, P = 0.002). The MLSStrad speed and MLSSsingle speed were poorly correlated (r = 0.29, P = 0.49). CONCLUSION: The single-visit method of determining the MLSS substantially underestimates the actual MLSS. PMID- 16260975 TI - Estrogen replacement, muscle composition, and physical function: The Health ABC Study. AB - PURPOSE: Although the beneficial effects of estrogen use on cardiovascular and cognitive function in postmenopausal women have been recently discredited, controversy remains regarding its usefulness for maintaining skeletal muscle mass or strength. Therefore, the purpose of this study was to determine whether estrogen use is associated with enhanced muscle composition and, if so, whether this translates into improved strength and physical function. METHODS: Cross sectional analysis of 840 well-functioning community-dwelling white women (current estrogen replacement therapy (ERT) users = 259, nonusers = 581) aged 70 79 yr participating in the Health, Aging and Body Composition Study. Muscle composition of the midthigh by computed tomography included cross-sectional area (CSA) of the quadriceps, hamstrings, intermuscular fat and subcutaneous fat, and muscle attenuation in Hounsfield units (HU) as a measure of muscle density. Isometric hand grip and isokinetic knee extensor strength were assessed by dynamometry. Physical function was assessed using a summary scale that included usual 6-m walk and narrow walk speed, repeated chair stands, and standing balance. RESULTS: In analyses of covariance adjusted for relevant confounders, quadriceps muscle CSA and HU were greater in current ERT than non-ERT women (P < 0.05). Grip strength was also greater (P < 0.05) in women taking ERT while knee extensor strength approached significance (P < 0.10). However, differences in muscle composition and strength were modest at < or =3.3%. There was no difference by ERT status for the hamstring muscles, fat CSA, or for physical function. CONCLUSION: The associations between ERT and muscle composition and strength were minor and did not translate into improved physical function. Initiation of ERT for preservation of muscle composition and function may not be indicated. PMID- 16260976 TI - Correlates of physical activity among pregnant women in the United States. AB - INTRODUCTION: Physical activity recommendations for pregnant and nonpregnant women have been issued, but little data exist to compare the extent that these women are meeting the recommendation levels. METHODS: A population-based, cross sectional study using data from the 1994, 1996, 1998, and 2000 Behavioral Risk Factor Surveillance System evaluated the physical activity recommendations met by pregnant and nonpregnant women. The study population included women who were 18 44 yr of age. Random digit dialing, telephone surveys were used to assess all information. Pregnant and nonpregnant women were categorized into five mutually exclusive groups based upon their level of physical activity in the past month: vigorous activity (meeting guidelines) moderate activity (meeting guidelines), moderate or vigorous activity (not meeting guidelines), irregular activity, or no physical activity. RESULTS: Nonpregnant women were more likely than pregnant women to meet the vigorous or moderate physical activity recommendations. Walking was the most common activity among pregnant and nonpregnant women (52 and 45%, respectively). Pregnant women meeting the moderate or vigorous physical activity recommendations were more likely to be younger, nonhispanic white, more educated, not married, nonsmokers, and to have higher incomes. CONCLUSION: Many pregnant women do not appear to be meeting the physical activity recommendations. Healthcare providers should further encourage the promotion of physical activity during uncomplicated pregnancies. PMID- 16260977 TI - Video analysis of the deformation and effective mass of gymnastics landing mats. AB - INTRODUCTION: Landing mats that undergo large area deformation are now essential for the safe completion of landings in gymnastics. The aim of this study was to determine the effective mass, shock transmission time, and deformation characteristics of a mat during impacts using high-speed video and to improve the accuracy of measuring foot-mat contact forces during landing. For validation, the same variables need to be accurately assessed using accelerometer and force plate data. METHODS: A 24-kg impactor with an attached accelerometer was dropped onto the sample mat from various heights. The surface deformation of the mat was recorded using high-speed video, and force data were obtained from a force plate beneath the mat. RESULTS: The maximum vertical deformations were between 0.088 and 0.118 m, with corresponding volume deformation estimates ranging from 0.030 to 0.044 m3. The delay between accelerometer and force plate readings at initial contact was approximately 7 ms. The delay between peak acceleration and peak force was 3 ms. The peak acceleration calculated from the video data was within 2.5% of that recorded via the accelerometer. The effective mass of the mat being accelerated corresponded to forces that ranged from 481 to 930 N, up to 12% of the peak force. CONCLUSIONS: The acceleration estimates obtained from high-speed video were combined with the effective mass estimates from the volume calculation to give peak calculated forces at the bottom of the mat to within -1.1 to +3.7% of the force recorded via the force plate. The use of high-speed video can be used to give data of sufficient accuracy for measuring foot/mat contact forces in gymnastics landings. PMID- 16260978 TI - Comparison of PAEE from combined and separate heart rate and movement models in children. AB - PURPOSE: Accurate measurement of physical activity in children is a challenge. Combining physiological (e.g., heart rate (HR)) and body movement registration (e.g., accelerometry) may overcome limitations with either method used alone. This study aimed to compare the estimated physical activity energy expenditure (PAEE) from hip- and ankle-mounted MTI Actigraphs, a hip-mounted Actical, and a new combined HR and movement sensor, the Actiheart (Cambridge Neurotechnology, Papworth, UK). METHODS: Resting EE and submaximal EE (treadmill walking and running) were measured in 39 children (13.2 +/- 0.3 yr) by indirect calorimetry during a progressive treadmill exercise bout. Associations between monitor outputs (activity counts, HR, and activity counts + HR) and the criterion were examined by linear regression models. The agreement between measured and predicted PAEE was examined by modified Bland-Altman plots in a subsample of participants. RESULTS: The combined Actiheart model (activity counts + HR) had the strongest relationship with PAEE (R2 = 0.86), compared with those from the single-measure models (R2 = 0.69 and 0.82 for the activity model and HR model). The explained variances from the other activity monitors were lower (R2 = 0.50, 0.37, and 0.67) for the hip MTI, ankle MTI, and Actical, respectively. In cross validation analyses, significant correlations were observed between estimation errors of the methods with the criterion (r = 0.49 to 0.90) in all models using only activity counts indicating a large systematic error. The HR and combined models indicated less systematic error (r = 0.41 and 0.33, respectively). CONCLUSIONS: Of the techniques considered, combined HR and movement sensing is the most valid for estimating PAEE in children during treadmill walking and running, compared with movement or HR alone. It also has the lowest level of systematic error. PMID- 16260979 TI - Effect of carbohydrate and prolonged exercise on affect and perceived exertion. AB - INTRODUCTION: It has been reported that perceptions of exertion are attenuated during prolonged cycle exercise, following CHO ingestion. However, no studies to date have examined the influence of such feedings on psychological affect during prolonged exercise, even though affect and perceived exertion are different constructs. PURPOSE: To examine the influence of regular CHO beverage ingestion on affect (pleasure-displeasure) and perceived exertion during prolonged cycle exercise. METHODS: In a randomized, double-blind, counterbalanced design, nine endurance trained males cycled for 2 h at 70% VO2max on two occasions, separated by 1 wk. On each occasion, they consumed either a water placebo (PLA) or a 6.4% carbohydrate-electrolyte solution (CHO) immediately before they cycled (5 mL x kg(-1) body mass) and every 15 min thereafter (2 mL x kg(-1) body mass). Pleasure displeasure was assessed before, during, and after the prolonged bout of cycling. RESULTS: During exercise, reported pleasure initially improved and was subsequently maintained in the CHO trial, in contrast to a decline reported in the PLA trial. Ratings of pleasure-displeasure were more positive during recovery in the CHO trial compared with the PLA trial (P < 0.05) and the only significant increase (P < 0.05) in pleasure occurred 15 min postexercise in the CHO trial only. RPE increased (P < 0.05) over the course of the bout of cycling and was lower (P < 0.05) 75 min into exercise in the CHO trial. Immediately postexercise, plasma glucose concentration was higher in the CHO compared with the PLA trial (P < 0.05). A main effect of trial was found for plasma cortisol concentration, with higher values reported in PLA trial. CONCLUSION: Results suggest that CHO ingestion enhanced feelings of pleasure during and following prolonged cycling and highlighted the importance of assessing not only "what," but also "how" a person feels. PMID- 16260980 TI - Profile of physical activity levels in community-dwelling older adults. AB - PURPOSE: To examine relationships between selected sociodemographic, health related and environmental factors and levels of physical activity in older adults across three age groups. METHODS: Seven hundred sixty-four older adults (mean age = 77.4 +/- 8.6 yr) from a midsize Canadian city completed a self-administered questionnaire under researcher supervision. Level of physical activity was determined using the Physical Activity Scale for the Elderly (PASE). Correlates of physical activity were examined using previously validated questionnaires. The findings pertaining to personal and environmental factors are presented. RESULTS: Overall, significantly higher mean PASE scores were seen in those individuals in the following categories: male (P < 0.001), married or common-law (P < 0.001), not living alone (P < 0.001), not living in senior's housing (P < 0.001), higher levels of education (P < 0.001) and higher incomes (P < 0.001). Better physical health showed significant positive associations (P < 0.001) with PASE score. Individuals reporting at least four or more chronic health conditions had significantly lower PASE scores than those reporting no chronic conditions (P < 0.001). Significantly lower PASE scores were also reported in those using domestic services (P < 0.001). Higher PASE scores were related to the presence of hills, biking and walking trails, street lights, various recreation facilities, seeing others active and unattended dogs (P < 0.001 to P < 0.05). CONCLUSION: An understanding of the factors that influence physical activity behavior in older adults is critical to developing effective intervention strategies that will address the problem of physical inactivity in this population, and in doing so, improve the health status and quality of life of the older adult, while having a significant impact on healthcare expenditures. PMID- 16260981 TI - Are there limits to running world records? AB - PURPOSE: Previous researchers have adopted linear models to predict athletic running world records, based on records recorded throughout the 20th century. These linear models imply that there is no limit to human performance and that, based on projected estimates, women will eventually run faster than men. The purpose of this article is to assess whether a more biologically sound, flattened "S-shaped" curve could provide a better and more interpretable fit to the data, suggesting that running world records could reach their asymptotic limits some time in the future. METHODS: Middle- and long-distance running world record speeds recorded during the 20th century were modeled using a flattened S-shaped logistic curve. RESULTS: The logistic curves produce significantly better fits to these world records than linear models (assessed by separating/partitioning the explained variance from the logistic and linear models using ANOVA). The models identify a slow rise in world-record speeds during the early year of the century, followed by a period of "acceleration" in the middle of the century (due to the professionalization of sport and advances in technology and science), and a subsequent reduction in the prevalence of record-breaking performances towards the end of the century. The model predicts that men's world records are nearing their asymptotic limits (within 1-3%). Indeed, the current women's 1500-m world record speed of 6.51 m x s(-1) may well have reached its limit (time 3:50.46). CONCLUSIONS: Many of the established men's and women's endurance running world records are nearing their limits and, consequently, women's world records are unlikely to ever reach those achieved by men. PMID- 16260983 TI - The effect of training on running economy and performance in recreational athletes. AB - PURPOSE: To analyze the effect of an 8-wk training program on the energy cost of running (C) and the performance of 16 recreational males. METHODS: A training group (TG, N = 8, 25.3 +/- 2.9 yr, 183.6 +/- 7.3 cm, 80.9 +/- 9.6 kg) and a control group (CG, N = 8, 24.3 +/- 3.7 yr, 179.3 +/- 6.1 cm, 75.5 +/- 8.0 kg) performed three two-stage tests (TST) at weeks 0, 4, and 8 (W0, W4, W8). Speeds of the first (v-slow) and second stage (v-fast) were 2.4 +/- 0.3 vs 2.5 +/- 0.4 m x s(-1) and 3.7 +/- 0.3 vs 3.9 +/- 0.4 m.s (TG vs CG), respectively. Maximum running time at v-fast (T) served as the measure of performance. C was calculated from oxygen uptake above rest, blood lactate concentration, and speed. The TG trained 3-5x wk(-1) at an HR of +/-10 beats of the HR measured at v-slow at W0 (161 +/- 12 bpm). The CG did not train. RESULTS: At W0, there were no significant differences between the groups in T (377 +/- 47 vs 335 +/- 34 s) and C (v-slow: 4.1 +/- 0.3 vs 4.3 +/- 0.4 J x kg(-1) x m(-1); v-fast: 4.2 +/- 0.4 vs 4.0 +/- 0.4 J x kg(-1) x m(-1)). In the CG, T and C remained almost unchanged at W4 (363 +/- 38 s, 4.0 +/- 0.4 J x kg(-1) x m(-1)) and at W8 (342 +/- 49 s, 4.0 +/- 0.3 J x kg(-1) x m(-1)). In the TG, T increased (P < 0.05) at W4 (469 +/- 45 s) and at W8 (591 +/- 109 s). At v-fast, also C increased (P < 0.05) at W8 (4.6 +/- 0.4 J x kg(-1) x m(-1)), whereas at v-slow, C decreased (P < 0.05) at W4 (3.7 +/- 0.4 J x kg(-1) x m(-1)) with no further change at W8 (3.7 +/- 0.4 J x kg(-1) x m(-1)). CONCLUSION: The training successfully increased running performance in terms of T. During the initial training period, C could be reduced at the speed predominantly used in training. However, at high running speeds, C may even increase if the corresponding running time is largely increased. PMID- 16260982 TI - Running 8000 m fast or slow: Are there differences in energy cost and fat metabolism? AB - PURPOSE: To compare energy requirements and substrate use for running a given distance fast versus slow under field conditions. METHODS: Ten males and four females who were moderately endurance trained (32 +/- 6 yr; 71 +/- 11 kg; body mass index: 22.7 +/- 2.3 kg x m(-2); VO2max: 62.0 +/- 6.0 mL x min(-1) x kg(-1); individual anaerobic threshold [IAT]: 13.7 +/- 1.4 km x h(-1)) performed an incremental running protocol to determine IAT. Subsequently, two 8000-m runs at 70 and 95% IAT were performed on separate days in randomized order on an indoor track. Energy expenditure (EE) was measured by means of a portable metabolic device (indirect calorimetry). A meaningful difference in EE was defined as >10%. RESULTS: EE was significantly greater during the 95% IAT run than during the 70% IAT run (2650 +/- 276 and 2554 +/- 348 kJ, respectively). However, this difference was only 3.8 +/- 4.8%. Including measurements up to 10 min postexercise, the difference rose to 5.1 +/- 4.7% (95% IAT: 2830 +/- 301 kJ; 70% IAT: 2692 +/- 368 kJ). There was no significant difference between the absolute amounts of fat oxidized during the runs (70% IAT: 26 +/- 5 g; 95% IAT: 20 +/- 5 g). During the 95% IAT run, significantly more carbohydrates were metabolized than during the 70% IAT run (108 +/- 14 and 90 +/- 5 g, respectively). The difference in EE between the two runs increased from the first to the third part of the running distance (first: no significance; second: P < 0.01; third: P < 0.001). CONCLUSION: Energy requirements for a commonly run distance in recreational endurance training differ significantly but not relevantly between slow and fast speeds. However, increasing total running distance might lead to larger differences. PMID- 16260984 TI - Laboratory measurement of posture allocation and physical activity in children. AB - PURPOSE: The purpose of this study was to validate the combined use of inclinometers and accelerometers to measure body posture and movement in children in a laboratory setting. METHODS: We performed two separate experiments. In the first experiment, we tested the hypothesis that four inclinometers (tilt sensors) could be used to capture body posture in children. We observed and recorded body posture in eight healthy children (mean +/- SD; body mass index (BMI), 18 +/- 3 kg x m(-2)) on 2880 occasions and compared these records with the inclinometer data. In the second experiment, the hypothesis was that two inclinometers could be used to determine whether 18 children (BMI, 21 +/- 5 kg x m(-2)) were sedentary. We observed and recorded sedentariness (sitting/lying compared to standing) on 5575 occasions and compared these records with the inclinometer data. In both of these experiments, we also addressed the hypothesis that accelerometer output, when measured at varying velocities, correlated with walking energy expenditure. RESULTS: In experiment 1, body posture was correctly identified in 2880 out of 2880 inclinometer measurements. In experiment 2, sedentary behavior was correctly identified in 5575 out of 5575 occasions. For the entire group, acceleration and body weight correlated well with energy expenditure (r2 = 0.84). CONCLUSION: The inclinometer-accelerometer system that we tested can be used to measure body posture and movement. We can measure sedentary behavior using two inclinometers instead of four inclinometers. This monitoring system may be useful for measuring energy expenditure, body posture, and physical activity in children. PMID- 16260985 TI - Effect of activity and age on long bones using a new densitometric technique. AB - INTRODUCTION: Long bone structural parameters such as cross-sectional area or area moments of inertia are useful measures of long bone mechanical properties. We implemented a three-scan densitometric method to measure structural parameters in long bones of the lower leg in vivo. The validated method was applied to investigate the relationship between activity level, age, and long bone structural parameters in women. METHODS: An aluminum phantom was used to estimate in vivo setup accuracy. In vivo precision was determined by same-day repeated measures on human subjects. For the activity study, women were recruited in two age groups (25-35, 60+yr) and two activity levels (recreational runners, nonrunners). Scans were taken of the middle third of the lower right leg; structural parameters for the tibia and fibula were determined at each scan line, averaged over the section, and adjusted by factors accounting for body size variations. RESULTS: Aluminum phantom cross-sectional area was underestimated by 4-6%, principal moments were underestimated by <5%, and principal angles were within +/-1.2 degrees. In vivo precision results (lower energy, scans spanning 60 degrees) indicated coefficients of variation for cross-sectional area (A), principal moments of inertia (Imax, Imin), and polar moment of inertia (J) of 0.52, 5.87, 2.22, and 3.82%, respectively. The activity study showed mean adjusted tibial A, Imax, Imin, and J were significantly higher in runners compared with nonrunners. There was no dependence on age. CONCLUSIONS: A three scan densitometric method for measuring cross-sectional structural parameters in long bones in vivo was validated; accuracy and precision measurements establishes confidence limits. From the activity study results, we postulate that higher loads associated with running lead to increased cross-sectional parameters to support axial loads, bending, and torsion in the tibia. PMID- 16260986 TI - Aquatic plyometric training increases vertical jump in female volleyball players. AB - PURPOSE: Numerous studies have reported that land-based plyometrics can improve muscular strength, joint stability, and vertical jump (VJ) in athletes; however, due to the intense nature of plyometric training, the potential for acute muscle soreness or even musculoskeletal injury exists. Performance of aquatic plyometric training (APT) could lead to similar benefits, but with reduced risks due to the buoyancy of water. Unfortunately, there is little information regarding the efficacy of APT. Thus, the purpose of this study was to examine the effects of APT on VJ and muscular strength in volleyball players. METHODS: Nineteen female volleyball players (aged 15 +/- 1 yr) were randomly assigned to perform 6 wk of APT or flexibility exercises (CON) twice weekly, both in addition to traditional preseason volleyball training. Testing of leg strength was performed at baseline and after 6 wk, and VJ was measured at baseline and after 2, 4, and 6 wk. RESULTS: Similar increases in VJ were observed in both groups after 4 wk (APT = 3.1%, CON = 4.9%; both P < 0.05); however, the APT group improved by an additional 8% (P < 0.05) from week 4 to week 6, whereas there was no further improvement in the CON group (-0.9%; P = NS). After 6 wk, both groups displayed significant improvements in concentric peak torque during knee extension and flexion at 60 and 180 degrees x s(-1) (all P < 0.05). CONCLUSIONS: The combination of APT and volleyball training resulted in larger improvements in VJ than in the CON group. Thus, given the likely reduction in muscle soreness with APT versus land-based plyometrics, APT appears to be a promising training option. PMID- 16260987 TI - Should time trial performance be predicted from three serial time-to-exhaustion tests? PMID- 16260989 TI - Mathematical constants that vary? PMID- 16260993 TI - The development of a patient safety program across the continuum of care. AB - In keeping with the national emphasis on patient safety, and in keeping with the concept that quality healthcare is not episodic but spans the entire continuum of care, with the patient's home being the "preferred site of care," the Home Operations Utilizing Safety Education Program was developed. This program uses patient assessment, education, and other interventions that adjust the home environment to complement the patient's needs. The assessment and education serve as a link between in-patient, ambulatory care, and the patient's home in a cost effective and efficient manner for reducing preventable injuries in a predominantly rural setting. The unique aspects of this program include patient follow-up in the home after initial face-to-face interviews/assessments and interventions via a hand-held camera phone. This program was developed as part of our plan to further enhance safety as a cultural norm within our Medical Center. PMID- 16260994 TI - The transforming effect of handheld computers on nursing practice. AB - Handheld computers have the power to transform nursing care. The roots of this power are the shift to decentralization of communication, electronic health records, and nurses' greater need for information at the point of care. This article discusses the effects of handheld resources, calculators, databases, electronic health records, and communication devices on nursing practice. The US government has articulated the necessity of implementing the use of handheld computers in healthcare. Nurse administrators need to encourage and promote the diffusion of this technology, which can reduce costs and improve care. PMID- 16260995 TI - Information technology: changing nursing processes at the point-of-care. AB - Changing societal demographics, increasing complexity in healthcare knowledge, and increasing nursing shortages have led healthcare strategists to call for a redesign of the healthcare system. Embedded within most redesign recommendations is the increased use of technology to make nursing practice more efficient. However, information technology (IT) has the potential to go beyond simple efficiency increases. If IT is perceived truly as a part of the redesign of healthcare delivery rather than simply the automation of existing processes, then it can change nursing processes within institutions and furthermore change the point-of-care between nurses and patients. Nursing adoption of technology within the workplace is a result of the interactions between technical skills, social acceptance, and workplace culture. Nursing needs for information not only influence their adoption of particular technologies but also shape their design. The objective of this article is to illustrate how IT can change not only nursing practice and processes but also the point-of-care. A case study of the use of IT by nurses in telehomecare is presented and administrative implications are discussed. PMID- 16260996 TI - Cultivating informatics competencies in a community of practice. AB - To move the healthcare industry into the 21st century, nurses must become savvy in the use of informatics to provide optimal care to their patients. However, the relatively few formal nursing informatics programs that exist across the country are simply not adequate to meet the demands of both new and existing nurses. Informatics competencies must be incorporated into nursing curricula at entry level and via staff development to provide a ready workforce. Creative faculty development strategies that capitalize on the concept of faculty as a community of practice are required to incorporate informatics competencies into nursing curricula. PMID- 16260997 TI - Aging well with smart technology. AB - As baby-boomers age, the need for long-term nursing care services increases. In the future, there will simply not be enough long-term care facilities to accommodate all of these patients. In addition, many people prefer to grow old at home, a concept known as aging-in-place. Smart home technology facilities aging in-place by assisting patients with emergency assistance, fall prevention/detection, reminder systems, medication administration and assistance for those with hearing, visual or cognitive impairments. Benefits include making aging-in-place a reality, continuous monitoring, and improved psychosocial effects. Concerns of this technology include cost, availability of technology, retrofitting complications, and potential inappropriate use of the technology. Overall, the concept of smart homes is gaining in popularity and will expand the role of the nurse in the future. It is important for all nurses to understand how their practices will be transformed as smart homes become a reality for the aging population. PMID- 16260998 TI - Changing the face of standard nursing practice through telehealth and telenursing. PMID- 16260999 TI - From Tele-ed to Telehealth: the need for IT ubiquity in nursing. AB - Although numerous studies have shown that information technology (IT) improves care, makes the workplace better for clinicians, and reduces costs, healthcare especially nursing-has been slow to adopt it for a variety of reasons. However, because of a worsening nursing shortage, couples with increasing healthcare costs and administrative overhead, nursing demands that IT become not only common, but pervasive, playing a role in every aspect of nursing, from recruitment and education, to patient care. This article will examine the several potential applications of IT to nursing and their benefits, challenges to widespread IT adoption, and the importance of IT ubiquity to the very viability of the profession. PMID- 16261000 TI - Ethics in informatics: the intersection of nursing, ethics, and information technology. PMID- 16261101 TI - A comparison of pedicle and lateral mass screw construct stiffnesses at the cervicothoracic junction: a biomechanical study. AB - STUDY DESIGN: Biomechanical comparison of five different posterior cervicothoracic junction (C7-T1) fixation constructs in a cadaveric model. OBJECTIVES: To determine whether augmenting C7 lateral mass screws with spinous process wires or additional fixation in the C6 lateral mass can create constructs of similar normalized stiffness to that of C7 pedicle screws. SUMMARY OF BACKGROUND DATA: Cervical pedicle screws are known to provide excellent fixation but are potentially dangerous and technically demanding to insert. Lateral mass screws are safer and easier to insert but have less pullout strength and must often be short at C7. METHODS: Twelve cadaveric cervicothoracic specimens (C5-T2) were randomly assigned to one of three experiments: Experiment A (Part 1 and Part 2), Experiment B, and Experiment C (Part 1 and Part 2) (n = 4 each for each experiment). First, the intact specimens were biomechanically tested according to a seven-part loading protocol. The specimens were then destabilized, and then restabilized with the following constructs in conjunction with bilateral T1 pedicle screws and biomechanically tested again using the same seven-part biomechanical protocol as was applied to the intact specimens. Experiment A: Part 1: lateral mass screw fixation at C7 (C7LM); then Part 2: retested after augmentation with triple wiring (C7LM+W). Experiment B: pedicle screw fixation at C7 (C7PS). Experiment C: Part 1: C6 and C7 lateral mass screws (C6C7LM); then Part 2: retested after augmentation with triple wiring (C6C7LM+W). Thus, five different constructs were biomechanically compared in these three experiments. RESULTS: None of the lateral mass constructs demonstrated a significant increase in normalized stiffness when augmented with wiring in any mode of testing. In axial compression, the C7PS construct showed significantly higher (P < 0.001) normalized stiffness than any of the other four constructs. In extension, there were no significant differences among any of the five constructs. Inflexion, right/left lateral bending and right/left axial torsion, the C7PS construct again showed significantly higher normalized stiffness (P < 0.05) than lateral mass fixation at C7 alone. However, in these five modes of testing, the addition of a secondary point of lateral mass fixation at C6 (C6C7LM) produced a construct with a normalized stiffness similar to that of C7PS with no significant difference (P > 0.05). CONCLUSION: C7 pedicle screw fixation provides the construct with the highest normalized stiffness for stabilizing the cervicothoracic junction. If C7 pedicle fixation is not possible, then performing two-level lateral mass fixation at C6 and C7 will achieve a construct with similar normalized stiffness except in axial compression. The addition of triple wiring to the spinous processes does not significantly increase lateral mass construct normalized stiffness. PMID- 16261102 TI - A Cochrane review of electrotherapy for mechanical neck disorders. AB - STUDY DESIGN: Systematic review. OBJECTIVE: To assess whether electrotherapy relieves pain or improves function/disability in adults with mechanical neck disorders (MND). SUMMARY OF BACKGROUND DATA: The effectiveness of electrotherapy as a physiotherapy option has remained unclear. METHODS: Databases were searched from root to March 2003. Independent reviewers conducted selection, data abstraction, and quality assessment. Relative risk and standard mean differences were calculated. RESULTS: Fourteen comparisons were included. For the pain outcome, we found limited evidence of benefit, ie, pulsed electromagnetic field (PEMF) therapy resulted in only immediate post-treatment pain relief for chronic MND and acute whiplash (WAD). Other findings included unclear or conflicting evidence (Galvanic current for acute or chronic occipital headache; iontophoresis for acute, subacute WAD; TENS for acute WAD, chronic MND; PEMF for medium- or long-term effects in acute WAD, chronic MND); and limited evidence of no benefit (diadynamic current for reduction of trigger point tenderness in chronic MND, cervicogenic headache; permanent magnets for chronic MND; electrical muscle stimulation (EMS) for chronic MND). CONCLUSIONS: In pain as well as other outcomes, the evidence for treatment of acute or chronic MND by different forms of electrotherapy is either lacking, limited, or conflicting. PMID- 16261103 TI - Effects of backpack loading on the pulmonary capacities of normal schoolgirls and those with adolescent idiopathic scoliosis. AB - STUDY DESIGN: A prospective evaluation of the effects of backpack carriage on the pulmonary function of schoolgirls without spinal deformity versus those with adolescent idiopathic scoliosis (AIS). OBJECTIVE: To establish if recommended backpack load limits for normal schoolchildren are also appropriate for study participants with AIS. SUMMARY OF BACKGROUND DATA: The weight of schoolchildren's backpacks are of concern because of effects including compromise of pulmonary function. Impaired pulmonary function is also found with AIS, but the effect of backpack carriage on the respiratory parameters of schoolchildren with AIS has not previously been examined. METHODS: Forced vital capacity (FVC), forced expiratory volume (FEV1), peak expiratory flow (PEF), and forced expiratory flow (FEF25-75%) were recorded in 17 girls (mean age, 12 years) with moderate AIS (Cobb angle, 26 degrees-50 degrees) and 18 girls (mean age, 11 years) without musculoskeletal deformity during carriage of a backpack loaded at 0%, 5%, 7.5%, 10%, 12.5%, and 15% body weight in random order. Absolute values and proportions of reference values were analyzed by repeated-measures analysis of variance. RESULTS.: No interaction between load and group was found, indicating that backpack loading has a similar effect on the pulmonary function of both normal and AIS groups. However, all recorded pulmonary parameters were found to be significantly lower in the AIS than normal group, significantly so for the referenced FVC and PEF. A significant decrease in FVC and FEV1 was found with increasing backpack load, and the load at which these changes were found to be significant was lower than those established in previous studies. CONCLUSIONS: Pulmonary function may be more sensitive to backpack load than previously considered, especially when study participants with AIS are being considered, and the recommended loading limit of 10% body weight may not be applicable to schoolgirls with AIS. PMID- 16261104 TI - Adjacent segment degeneration at T1-T2 presenting as chest pain: case report. AB - STUDY DESIGN: A case report of a T1-T2 herniated disc adjacent to a cervical spine fusion. OBJECTIVE: To describe the clinical presentation and treatment of a symptomatic T1-T2 disc herniation adjacent to an anterior cervical spine arthrodesis. SUMMARY OF BACKGROUND DATA: Symptomatic adjacent segment disease after cervical spine arthrodesis occurs most commonly in the lower cervical spine motion segments and has not been previously reported at T1-T2. METHODS: A 51-year old female patient presented 4 years after three-level anterior cervical discectomy and fusion involving the C5-T1 levels. She complained of chronic chest pain and had undergone extensive medical evaluation before presentation. Magnetic resonance imaging revealed a left-sided T1-T2 herniated disc compressing the T1 nerve root. She underwent T1-T2 anterior discectomy and fusion. RESULTS: The patient's symptoms resolved completely. CONCLUSION: Adjacent-segment degeneration after cervical spine fusion may rarely occur even at T1-T2, and the unusual symptoms of a T1 radiculopathy may obscure the diagnosis. The combination of sharp anterior paramedian chest pain and numbness or weakness of the hand should prompt consideration of the diagnosis when cardiac ischemia or other more serious conditions have been ruled out. PMID- 16261105 TI - Atraumatic vertebral artery dissection after cervical corpectomy: a traction injury? AB - STUDY DESIGN: Case report with review of the literature. OBJECTIVES: Presented is the first case of vertebral artery dissection secondary to intraoperative traction in cervical spine surgery. The pathogenesis and management of vertebral dissection in the immediate postoperative period are reviewed in detail. SUMMARY OF BACKGROUND DATA: Vertebral artery dissection is commonly associated with direct trauma, atraumatic or spontaneous. There are numerous reports of direct injury to the vertebral artery with cervical spine surgery, and it is a well recognized risk. Cervical traction is also used routinely for improved placement of intervertebral devices/grafts. This is the first report of vertebral artery dissection occurring intraoperatively secondary to traction. The postoperative management is reviewed in detail. METHODS: Case study with extensive review of the literature. RESULTS: The patient underwent C6 corpectomy without intraoperative complications. Intraoperative wake-up test was normal. The patient remained intubated overnight for airway precautions. On postoperative day 1, the patient was lethargic and not following commands. Emergent CT of the brain and cervical spine revealed multiple posterior circulation infarcts with normal cervical spine and no hematoma. A stat angiogram revealed vertebral dissection. Medical management was initially attempted; however, infarcts continued, eventually requiring posterior fossa craniectomy/decompression and sacrificing the vertebral at the O-A junction. CONCLUSIONS: This is the first report of vertebral artery dissection occurring secondary to traction in cervical spine surgery. Surgeons must be aware that traction, even when performed appropriately, is not without risks. Anomalous vertebral arteries, osteophytes, and numerous other anatomic variants can lead to vertebral injury with traction. PMID- 16261106 TI - Lumbar vertebral hemangioma causing cauda equina syndrome: a case report. AB - STUDY DESIGN: Case report. OBJECTIVES: To report a case of lumbar hemangioma causing neurogenic claudication and early cauda equina, managed with hemostatic vertebroplasty and posterior decompression. SUMMARY OF BACKGROUND DATA: This is the first report to our knowledge of a lumbar hemangioma causing neurogenic claudication and early cauda equina syndrome. Most hemangiomas causing neurologic symptoms occur in thoracic spine and cause spinal cord compression. Vertebroplasty as a method of hemostasis and for providing mechanical stability in this situation has not been discussed previously in the literature. METHODS: L4 hemangioma was diagnosed in a 64-year-old woman with severe neurogenic claudication and early cauda equina syndrome. Preoperative angiograms showed no embolizable vessels. Posterior decompression was performed followed by bilateral transpedicular vertebroplasty. The patient received postoperative radiation to prevent recurrence. RESULTS: Complete relief of neurogenic claudication and cauda equina with less than 100 mL of blood loss. CONCLUSION: A lumbar hemangioma of the vertebral body, although rare, can cause neurogenic claudication and cauda equina syndrome. Intraoperative vertebroplasty can be an effective method of hemostasis and provide stability of the vertebra following posterior decompression. PMID- 16261108 TI - A prospective randomized study of preemptive analgesia for postoperative pain in the patients undergoing posterior lumbar interbody fusion: continuous subcutaneous morphine, continuous epidural morphine, and diclofenac sodium. AB - STUDY DESIGN: A prospective randomized clinical trial in preemptive analgesia for postoperative pain was conducted. OBJECTIVE: To compare the efficacy of three preemptive analgesics combined with local anesthesia: continuous subcutaneous morphine (SC), continuous epidural morphine (ED), and diclofenac sodium (DS). SUMMARY OF BACKGROUND DATA: Systemic opioids are known to be effective methods of postoperative pain control. The use of epidural morphine for postoperative analgesia has been a standard treatment in spinal surgery. Only a few studies in the literature have investigated the efficacy of preemptive analgesia using morphine. This is the first prospective randomized clinical trial to assess both subcutaneous and epidural continuous administration of opioids for preemptive analgesia. METHODS: For this study, 73 patients were assigned randomly to one of three treatment groups: SC, ED, or DS. All patients underwent posterior lumbar interbody fusion with instrumentation. Pain management was assessed using the visual analogue scale (VAS). Usage of supplemental analgesics, the time to first request of them, and side effects were also investigated. RESULTS: Twenty-two patients were randomized to SC, 23 to ED, and 27 to DS. No baseline variable differences among the three groups were seen. The results showed no significant difference in analgesic effects among those three preemptive analgesics. Only immediately after surgery (at 0 hours), the VAS of the DS group was lowest among three groups. But the DS group took more supplemental analgesic drugs until 72 hours, and the time to first request of this group was shorter than that of the other two groups. High rates of minor side effects were seen in both the ED and DS groups. SC gave moderate analgesic effects as well as the other two groups with few adverse effects. CONCLUSIONS: DS provided a favorable effect immediately after surgery, but the effective time was short and the patient needed more supplemental drugs after that. ED did not give the expected effect, with comparatively high rates of side effects. Continuous epidural anesthesia did not seem to be suitable for preemptive analgesia. Continuous subcutaneous morphine brought some analgesic effects with a low rate of complications. It can be an attractive method for postoperative analgesia with technical ease. PMID- 16261109 TI - Corticosteroids peroperatively diminishes damage to the C-fibers in microscopic lumbar disc surgery. AB - STUDY DESIGN: A subgroup analysis of patient outcomes from a double-blind randomized controlled study comparing corticosteroid versus saline in microscopic lumbar disc surgery. OBJECTIVES: To study if the use of corticosteroids combined with surgery alleviates the damage to the nerve fibers in lumbar disc herniation. SUMMARY OF BACKGROUND DATA: The use of quantitative sensory testing can detect damage to the myelinated A-delta fibers (cold sense) and the unmyelinated C fibers (warmth sense). Corticosteroids combined with surgery in lumbar disc surgery enhance the outcome after surgery in terms of less pain and more rapid rehabilitation. METHODS: Analyzing quantitative sensory testing before surgery and after 2 weeks and 2 years. RESULTS: In the corticosteroid group, we saw a statistically significant normalization for the warmth disturbance comparing with control group, which not was detected concerning the cold disturbance. CONCLUSIONS: The use of corticosteroids combined with surgery seems to protect some of the damage to the C-fibers in lumbar disc herniation. PMID- 16261111 TI - A randomized, double-blind, controlled trial: intradiscal electrothermal therapy versus placebo for the treatment of chronic discogenic low back pain. AB - STUDY DESIGN: A prospective, randomized, double-blind, placebo-controlled trial of intradiscal electrothermal therapy (IDET) for the treatment of chronic discogenic low back pain (CDLBP). OBJECTIVES: To test the safety and efficacy of IDET compared with a sham treatment (placebo). SUMMARY OF BACKGROUND DATA: In North America alone, more than 40,000 intradiscal catheters have been used to treat CDLBP. The evidence for efficacy of IDET is weak coming from retrospective and prospective cohort studies providing only Class II and Class III evidence. There is one study published with Class I evidence. This demonstrates statistically significant improvements following IDET; however, the clinical significance of these improvements is questionable. METHODS: Patients with CDLBP who failed to improve following conservative therapy were considered for this study. Inclusion criteria included the presence of one- or two-level symptomatic disc degeneration with posterior or posterolateral anular tears as determined by provocative computed tomography (CT) discography. Patients were excluded if there was greater than 50% loss of disc height or previous spinal surgery. Fifty-seven patients were randomized with a 2:1 ratio: 38 to IDET and 19 to sham procedure (placebo). In all cases, the IDET catheter was positioned to cover at least 75% of the annular tear as defined by the CT discography. An independent technician connected the catheter to the generator and then either delivered electrothermal energy (active group) or did not (sham group). Surgeon, patient, and independent outcome assessor were all blinded to the treatment. All patients followed a standard postprocedural rehabilitation program. Independent statistical analysis was performed. OUTCOME MEASURES: Low Back Outcome Score (LBOS), Oswestry Disability Index (ODI), Short Form 36 questionnaire (SF-36), Zung Depression Index (ZDI), and Modified Somatic Perceptions Questionnaire (MSPQ) were measured at baseline and 6 months. Successful outcome was defined as: no neurologic deficit, improvement in LBOS of greater then 7 points, and improvement in SF-36 subsets (physical function and bodily pain) of greater than 1 standard deviation. RESULTS: Baseline demographic data, initial LBOS, ODI, SF-36, ZDI, and MSPQ were similar for both groups. No neurologic deficits occurred. No subject in either arm showed improvement of greater than 7 points in LBOS or greater than 1 standard deviation in the specified domains of the SF-36. Mean ODI was 41.42 at baseline and 39.77 at 6 months for the IDET group, compared with 40.74 at baseline and 41.58 at 6 months for the placebo group. There was no significant change in ZDI or MSPQ scores for either group. CONCLUSIONS: The IDET procedure appeared safe with no permanent complications. No subject in either arm met criteria for successful outcome. Further detailed analyses showed no significant change in outcome measures in either group at 6 months. This study demonstrates no significant benefit from IDET over placebo. PMID- 16261113 TI - Differentiation of mesenchymal stem cells transplanted to a rabbit degenerative disc model: potential and limitations for stem cell therapy in disc regeneration. AB - STUDY DESIGN: An in vivo study to assess the differentiation status of mesenchymal stem cells (MSCs) transplanted to the nucleus pulposus of degenerative discs in a rabbit model. OBJECTIVES: To evaluate the fate of MSCs transplanted to the nucleus pulposus of degenerative discs in a rabbit and to determine whether they are a suitable alternative for cell transplantation therapy for disc degeneration. SUMMARY OF BACKGROUND DATA: Although MSCs have been proposed as candidate donor cells for transplantation to treat intervertebral disc degeneration, their differentiation after transplantation has not been adequately investigated. METHODS: Autologous MSCs, labeled with green fluorescent protein, were transplanted into mature rabbits. Consecutive counts of transplanted MSCs in the nucleus area were performed for 48 weeks after transplantation. Differentiation of transplanted cells was determined by immunohistochemical analysis. The proteoglycan content of discs was measured quantitatively using a dimethylmethylene blue assay, and mRNA expression of Type I and II collagen, aggrecan and versican was measured semi-quantitatively using reverse transcription polymerase chain reaction. RESULTS: Many cells that were positive for green fluorescent protein were observed in the nucleus pulposus of cell-transplanted rabbit discs 2 weeks after transplantation. Their number increased significantly by 48 weeks. Some GFP-positive cells were positive for cell-associated matrix molecules, such as Type II collagen, keratan sulfate, chondroitin sulfate, aggrecan, and the nucleus pulposus phenotypic markers, hypoxia inducible factor 1 alpha, glutamine transporter 1, and matrix metalloproteinase 2. MSCs did not show significant expression of these molecules before transplantation. Biochemical and gene expression analyses showed significant restoration of total proteoglycan content and matrix-related genes compared with nontransplanted discs. CONCLUSIONS: MSCs transplanted to degenerative discs in rabbits proliferated and differentiated into cells expressing some of the major phenotypic characteristics of nucleus pulposus cells, suggesting that these MSCs may have undergone site-dependent differentiation. Further studies are needed to evaluate their functional role. PMID- 16261114 TI - Spinal kyphosis causes demyelination and neuronal loss in the spinal cord: a new model of kyphotic deformity using juvenile Japanese small game fowls. AB - STUDY DESIGN: Histologic changes in the spinal cord caused by progressive spinal kyphosis were assessed using a new animal model. OBJECTIVES: To evaluate the effects of chronic compression associated with kyphotic deformity of the cervical spine on the spinal cord. SUMMARY OF BACKGROUND DATA: The spinal cord has remarkable ability to resist chronic compression, however, delayed paralysis is sometimes seen following the development of spinal kyphosis. In the past, no animal model to clarify the mechanism of spinal cord damage due to spinal kyphotic deformity has been available. METHODS: Laminectomy and bilateral facetectomy at the C4-C5 level was performed in 52 Japanese small game fowls. Histologic changes in the spinal cord associated with progressive kyphotic deformity were examined at different time points after surgery in each animal. The degree of spinal cord flattening and the severity of demyelination in histologic sections were quantitatively evaluated using an image analyzer, and their association with the kyphotic angle was analyzed. Changes in the microvascular distribution in the spinal cord were also examined by microangiography. RESULTS: In all operated animals, progressive kyphosis developed reproducibly. The kyphotic angle increased gradually until 3 weeks after surgery and stabilized thereafter. There was a significant correlation between the kyphotic angle and the degree of spinal cord flattening. The spinal cord was compressed most intensely at the apex of the kyphosis, where demyelination of the anterior funiculus as well as neuronal loss and atrophy of the anterior horn were observed. Demyelination progressed as the kyphotic deformity became more severe, initially affecting the anterior funiculus and later extending to the lateral and then the posterior funiculus. Angiography revealed a decrease of the vascular distribution at the ventral side of the compressed spinal cord. CONCLUSIONS: Progressive kyphosis of the cervical spine resulted in demyelination of nerve fibers in the funiculi and neuronal loss in the anterior horn due tochronic compression of the spinal cord. These histologic changes seem to be associated with both continuous mechanical compression and vascular changes in the spinal cord. PMID- 16261115 TI - Ectopic firing due to artificial venous stasis in rat lumbar spinal canal stenosis model: a possible pathogenesis of neurogenic intermittent claudication. AB - STUDY DESIGN: An electrophysiologic analysis was performed on a chronic lumbar spinal stenosis model of rats. The effects of venous stasis on ectopic firing originating in the nerve root were investigated. OBJECTIVES: To elucidate the mechanisms of neurogenic intermittent claudication in lumbar spinal canal stenosis. SUMMARY OF BACKGROUND DATA: Neurogenic intermittent claudication has been known as a characteristic symptom of lumbar spinal canal stenosis (LSCS), but the pathogenesis is poorly understood. Venous stasis of cauda equina has been speculated as a possible factor in the development of symptoms of the lower extremities while walking. On the other hand, ectopic firing originating in the dorsal root ganglia is thought to play an important role in the development of radicular pain or abnormal sensation. However, a direct association between venous stasis and ectopic firing has been never demonstrated. METHODS: Using 10 Wistar rats, the LSCS group was prepared by inserting two silicone strips into the L3 and L5 dorsal epidural spaces. Another 10 animals were treated without silicone insertion as a sham group. Fourteen days later, the ectopic firing originating in the L5 nerve root was antidromically recorded from the distal stump of the severed sural nerve. After recording initial spontaneous firing, the posterior vena cava was clamped for 60 seconds to simulate a transient venous stasis and the changes in firing were analyzed. RESULTS: None of the animals in the sham group showed a significant change in firing due to venous stasis. In contrast, most animals in the LSCS group showed a marked increase in firing during the venous stasis with some latency and then returned to the initial firing state after the release of the clamp. This phenomenon was repeated as long as the animals were maintained. CONCLUSIONS: We demonstrated that ectopic firing was elicited by venous stasis only in the LSCS animals. Therefore, the venous stasis may be a major factor of neurogenic intermittent claudication. PMID- 16261116 TI - Exposure to pulsed low intensity ultrasound stimulates extracellular matrix metabolism of bovine intervertebral disc cells cultured in alginate beads. AB - STUDY DESIGN: In vitro study on the effects of pulsed low intensity ultrasound on the cellular metabolism of bovine intervertebral disc cells. OBJECTIVE: To determine whether pulsed low intensity ultrasound has effects on cell proliferation and extracellular matrix metabolism by bovine intervertebral disc cells. SUMMARY OF BACKGROUND DATA: The application of pulsed low intensity ultrasound is known to be effective in stimulating fracture and cartilage repair. However, the effects of pulsed low intensity ultrasound on intervertebral disc cells are not known. METHODS: Cells of the nucleus pulposus and inner and outer anulus fibrosus were enzymatically isolated from bovine coccygeal tissue and precultured in alginate beads for 14 days. In the ultrasound group, pulsed low intensity ultrasound was administered to the culture for 20 minutes daily for an additional 20 days. The control group was cultured in the same way but without administration of ultrasound. Cell viability, DNA content, proteoglycan and collagen synthesis, and proteoglycan content at days 10 and 20 after the initiation of treatment were evaluated. Characterization of newly synthesized collagen and proteoglycan was performed. RESULTS: No significant differences in cell viability and DNA content were observed between the two groups. On day 20, proteoglycan synthesis was increased by the application of pulsed low intensity ultrasound in nucleus pulposus and inner and outer anulus fibrosus cells (24%-26% increase, P < 0.001). The application of pulsed low intensity ultrasound increased proteoglycan content in alginate beads containing inner and outer anulus fibrosus cells (P < 0.05). Collagen synthesis by cells isolated from all three zones of the intervertebral disc was increased by the application of pulsed low intensity ultrasound (16%-19% increase, P < 0.05-0.0001). CONCLUSIONS: The application of pulsed low intensity ultrasound stimulated extracellular matrix metabolism in intervertebral disc cells. Pulsed low intensity ultrasound may prove useful for the physical stimulation of cell metabolism for tissue engineering of intervertebral disc tissue. PMID- 16261117 TI - Thoracic vertebral screw impingement on the aorta in an in vivo bovine model. AB - STUDY DESIGN: A bovine model was used to evaluate the effects of thoracic vertebral screw impingement of the aorta. OBJECTIVES: To evaluate the histologic and biomechanical changes in aortic wall tissue that was severely impinged by abutting instrumentation. SUMMARY OF BACKGROUND DATA: Case reports of vascular injury associated with spinal instrumentation generally describe intraoperative injury; some report delayed presentation of large vessel damage. Risks associated with placing instrumentation adjacent to large vessels are largely unknown. METHODS: Six 1-month-old calves underwent left-sided thoracotomies, exposing the anterior thoracic spine and aorta. With the heads removed, screws were inserted in reverse fashion into T6 through T11, leaving the screw tips 1 cm proud and abutting the aorta. After 3, 6, or 12 months (2 calves each), the spines were resected with the adjacent aorta and underwent radiographic, histologic, and biomechanical testing. RESULTS: Computed tomography revealed varying degrees of vessel impingement. Although there were no frank ruptures, 96% of aortic specimens showed histopathologic changes, including 52% with wall thinning; 43% were no longer impinged, yet 60% of these had increased collagen (scar). Biomechanical testing of screw-impinged aortas demonstrated a lower failure stress (1.2 +/- 0.5 N/mm vs. 1.8 +/- 0.4 N/mm, P = 0.016) but no difference in failure strain (42 +/- 9% vs. 32 +/- 10%, P = 0.06) than controls. CONCLUSIONS: Major impingement of vertebral screws on the aorta caused acute and chronic histopathologic and biomechanical changes in the vessel wall. This model represents a severe form of vessel penetration by a screw that confirms such a "worst case" scenario results in marked compromise of the vessel wall integrity. The sequelae of less severe impingement are unknown. PMID- 16261118 TI - Is posterior spinal cord shifting by extensive posterior decompression clinically significant for multisegmental cervical spondylotic myelopathy? AB - STUDY DESIGN: Posterior cervical spinal cord shifting after selective single laminectomy associated with partial laminotomies was compared with that after bilateral open-door laminoplasty between the C3 and C7 levels in relation to the clinical results of each procedure. OBJECTIVES: To investigate the clinical significance of posterior spinal cord shifting after extensive cervical laminoplasty. SUMMARY OF BACKGROUND DATA: Current techniques used for cervical laminoplasty for multisegmental cervical spondylotic myelopathy (CSM) are consecutively performed between the C3 and C6 or C7 levels with expectation that the spinal cord will shift backward to keep it clear of anterior compression. However, the clinical significance of the posterior spinal cord shifting remains controversial, and there has been no report verifying it by comparing limited posterior decompression procedures with conventional extensive ones. METHODS: Twenty-six patients with consecutive 2- to 3-level CSM who underwent selective laminoplasty (Group A) were enrolled in the study, and among 56 CSM patients who underwent bilateral open-door laminoplasty between the C3 and C7 levels, 25 who had consecutive 2- or 3- level stenosis identified by preoperative magnetic resonance imaging were used as controls (Group B). The recovery rate was calculated using preoperative and postoperative Japanese Orthopedic Association (JOA) scores for each patient, and for each patient's magnetic resonance imaging, the postoperative cervical curvature index was obtained according to Ishihara's method and the magnitude of postoperative backward shifting of the spinal cord was measured. RESULTS: There was no significant difference between the subjects in Groups A and B with respect to the spinal curvature index, preoperative JOA scores, and recovery rate, but the magnitude of the postoperative posterior shifting of the spinal cord was greater for those in Group B than for those in Group A. There was no correlation between the recovery rate and posterior shifting of the spinal cord for each group, and no correlation was also found between the curvature index and posterior shifting of the spinal cord. CONCLUSIONS: The outcome of posterior decompression surgery for multisegmental CSM is not correlated with the magnitude of postoperative backward shifting of the spinal cord. Extensive and consecutive decompression performed in conventional cervical laminoplasties is therefore not always necessary for multisegmental CSM. PMID- 16261119 TI - Cervical laminoplasty combined with muscle release in patients with athetoid cerebral palsy. AB - STUDY DESIGN: A retrospective study comparing cervical laminoplasty with or without muscle release for the treatment of cervical myelopathy resulting from athetoid cerebral palsy. OBJECTIVE: To assess the effectiveness of muscle release in the treatment of athetoid cerebral palsy. SUMMARY OF BACKGROUND DATA: While anterior and/or posterior spinal fusion has been generally accepted as necessary in surgical treatment for cervical myelopathy due to athetoid cerebral palsy, several studies have shown relatively favorable results following laminoplasty. Better results can be obtained by combining muscle release. METHODS.: Study participants were 10 patients who underwent cervical laminoplasty combined with muscle release (mean age, 44.6 years) and 15 patients who underwent cervical laminoplasty alone (mean age, 48.2 years). Therapeutic outcomes 1 year after surgery, as assessed by Kurokawa's methods and JOA scores, were compared between groups. RESULTS: Recovery rate 1 year after surgery was significantly higher for the muscle release group than for the control group. In both groups, recovery rates were significantly better for patients who could walk before surgery. CONCLUSIONS: Cervical laminoplasty combined with muscle release for the treatment of cervical myelopathy due to athetoid cerebral palsy is effective in simplifying postoperative therapy and improving JOA scores. PMID- 16261120 TI - An outcomes analysis of the treatment of cervical pseudarthrosis with posterior fusion. AB - STUDY DESIGN: A retrospective review of 33 consecutive patients treated with posterior fusion and selective nerve root decompression for the treatment of pseudarthrosis following anterior cervical discectomy and fusion. OBJECTIVES: Use standardized outcome measures to evaluate the results of posterior fusion with selective nerve root decompression as a treatment option for symptomatic pseudarthrosis of the cervical spine. SUMMARY OF BACKGROUND DATA: Pseudarthrosis after anterior cervical discectomy and fusion has been recognized as a cause of continued cervical pain and unsatisfactory outcomes. Debate continues as to whether a revision anterior approach or a posterior fusion procedure is the best treatment for symptomatic cervical pseudarthrosis. To our knowledge, standardized outcome measures have not been used to evaluate the results of either surgical treatment option; therefore, it is difficult to evaluate outcomes in these patients, let alone compare surgical treatment options. Data on fusion rates in these two surgical treatment groups suggest a trend of a higher fusion rate with utilization of a posterior revision procedure, but the largest study to date includes the study of only 19 patients treated with a posterior fusion. METHODS: Thirty-three consecutive patients with symptomatic pseudarthrosis following anterior cervical discectomy and fusion were treated with selective nerve root decompression and posterior fusion using iliac crest or local bone graft as well as posterior wiring and/or lateral mass plating. The average follow-up period was 46 months (range, 20-86 months). Patients were assessed using physical examination, flexion-extension lateral radiographs, and standardized outcome measures including the SF-36, Arthritis Impact Measurement Scales 2, and Cervical Spine Outcomes Questionnaire. RESULTS: All 33 patients (100%) demonstrated a solid fusion at their most recent follow-up, and all 33 patients noted significant improvement in their preoperative symptoms. No difference in fusion status was noted between those treated with iliac crest versus patients treated with local bone graft--all had a solid fusion; 72% of the patients were satisfied with the result of their surgery. Cervical Spine Outcomes Questionnaire pain scales demonstrated 52% of patients reported mild or nopain at follow-up, whereas 20% described their pain as "discomforting" and 28% of the patients continued to report moderate to severe pain. CONCLUSIONS: This is the first study to our knowledge to use standardized outcome measures to assess clinical outcome in patients treated with posterior fusion for pseudarthrosis after anterior cervical discectomy and fusion. Patients and surgeons need to understand the potential for success with this revision procedure but also be aware of the relatively high rate of continued moderate to severe pain observed in this patient population even after a solid fusion is achieved. All of the patients in this study fused with a single posterior fusion procedure, further supporting the relatively higher fusion rates observed in the literature using posterior fusion as a treatment for cervical pseudarthrosis. Our results also support the ability of surgeons to use local bone graft without iliac crest in a posterior fusion for cervical pseudarthrosis and therefore avoid the morbidity associated with iliac crest bone graft harvest. PMID- 16261121 TI - "Occult" osteoporotic vertebral fractures: vertebral body fractures without radiologic collapse. AB - STUDY DESIGN: Retrospective observational study. OBJECTIVES: We report our experience with patients who presented with osteoporotic vertebral fractures with no visible deformation of vertebral body. SUMMARY OF BACKGROUND DATA: The diagnosis of osteoporotic vertebral fractures largely relies on the observation of vertebral deformations on plain radiographs, termed vertebral collapses. There are no data on the characteristics, or indeed of the reality, of osteoporotic vertebral fractures with no significant deformation of the vertebral body. METHODS: We retrospectively analyzed cases that presented with acute back pain with no initial deformation of the vertebral body on plain radiographs, and later proved to be fresh osteoporotic vertebral body fractures. All cases met each of the following criteria: 1) The incriminated vertebra appeared normal on initial radiographs (Genant's Grade 0 deformation). 2) The diagnosis of fresh vertebral body fracture was confirmed by MRI. 3) The diagnosis of osteoporosis was made by the combination of established osteoporosis, ruling out of underlying disease, and follow-up. RESULTS: We observed 21 fractures in 16 patients (11 female/5 male; mean age, 72 years). Most of these fractures affected the lumbar spine (14 of 21 occurred at L2-L5). Osteoporosis was known beforehand in 9 patients and newly diagnosed in 7 patients. At follow-up, radiographs were obtained for 19 of 21 fractures: in 15 cases, the vertebral fracture developed a vertebral collapse (Genant's Grade > or = 0.5) in a mean of 12.5 weeks (range, 4-24 weeks); in the 4 remaining cases, the vertebra remained normal. All cases had a clinically favorable outcome. CONCLUSION: Osteoporotic vertebral fractures with no sign of vertebral collapse on initial radiographs do indeed exist. They are analogous to occult stress fractures well known in other skeletal sites. They must not be misdiagnosed as malignant lesions. PMID- 16261122 TI - Complete paraplegia following gunshot injury without direct trauma to the cord. AB - STUDY DESIGN: A retrospective review of all patients with neurologic deficit following gunshot wounds that did not penetrate the spinal canal. OBJECTIVES: To evaluate the possibility that neurologic deficit following gunshot wounds is possible without direct trauma to the cord. SUMMARY OF BACKGROUND DATA: Gunshot injuries to the spine account for 13% to 17% of all spinal cord injuries and are likely to result in complete paraplegia. Neurologic deficit is the result of direct trauma to the nervous tissue by the bullet, bone, or disc fragments, which compress the cord. METHODS: The medical charts, radiographs, CT scans, and myelographies or MRIs of all patients admitted to our hospital with neurologic deficit secondary to gunshot wounds between 1977 and 2003 were reviewed. Twenty six patients were identified: 15 with complete and 11 with incomplete paraplegia. In 19 patients, the spinal canal at the level of injury was explored and decompressed. RESULTS: In 3 patients with complete paraplegia at the thoracic level, CT scan, myelography (1 patient), MRI (2 patients), and operative exploration (2 patients) did not indicate any signs of canal compromise. Two were injured by low-velocity bullets passing through the vertebra (in 1 patient) and the posterior part of the lamina (in the second). The third was injured by high velocity bullets with no signs on MRI of any injury to the vertebral column or spinal cord. At follow-up 4.1 years later, on average, none had any neurologic recovery. CONCLUSION: Neurologic deficit following gunshot wounds is possible even without violation of the spinal canal. It is most probably the result of the kinetic energy emitted by the bullet. PMID- 16261123 TI - Spinal hydatidosis. AB - STUDY DESIGN: Retrospective study. OBJECTIVES: To revise the treatment of spinal hydatidosis. SUMMARY OF BACKGROUND DATA: About 50% of cases of bone hydatidosis are spinal. METHODS: Twenty patients with spinal hydatidosis were treated. There were 13 men and 7 women, with a mean age of 53.1 years. The median follow-up was 4.8 years. The infected area was the cervical spine in 1 patient, the dorsal spine in 7, the lumbar spine in 7, and the sacrum in 5. Curettage and resection of the infected bone were performed. The spine was exposed posteriorly in all but 2 patients. An additional posterior decompression and fusion with instrumentation were performed after removing the involved posterior elements in 8 patients. An anterior approach was used in 2 patients: a total corpectomy was performed and a bone graft was added. Two weeks later, a posterior decompression and fixation with instrumentation was done. No preoperative drugs were used. Medical antihelmintic treatment was used after surgery in all but 8 patients. RESULTS: Thirteen patients had spinal cord injury at the moment of surgery. The neurologic damage recovered only in 1 case. Repeated curettage was necessary in 13 cases: in 12 of them because of recurrence and in 1 because of surgical wound infection. In 6 cases of recurrence, there was also chronic wound infection. At last follow-up, 17 patients were free of disease. CONCLUSIONS: The treatment of choice of spinal hydatidosis is surgical removal of the affected vertebrae combined with posterior stabilization, followed by postoperative adjuvant chemotherapy. Recurrence is very frequent. PMID- 16261124 TI - Radiographic progression of lumbar spine disc degeneration is influenced by variation at inflammatory genes: a candidate SNP association study in the Chingford cohort. AB - STUDY DESIGN: A candidate gene association study in a longitudinal cohort. OBJECTIVE: To investigate the association between polymorphisms at 25 candidate genes and progression of individual radiographic features of lumbar disc degeneration (LDD). SUMMARY OF BACKGROUND DATA: LDD is characterized radiographically by the presence of osteophytes and disc space narrowing and is known to have a genetic component. Because of the high prevalence of radiographic features, progression may be a more useful phenotype clinically to study than prevalence itself. METHODS: We tested the effect on radiographic progression of LDD of polymorphisms in 25 genes, 24 of which had been previously tested with regards to knee osteoarthritis. The progression traits used were the change in radiographic grade over 9 years in osteophytes, disc space narrowing, and summary Kellgren-Lawrence grade. Lumbar spine radiographs (L1-L5) at baseline and at follow-up were read for 720 women genotyped at the 25 genes participating in the Chingford study. RESULTS: Polymorphisms in MMP3, TIMP1, and COX2, which encode molecules involved in inflammatory pathways, were associated with radiographic progression of LDD. The strongest associations observed (statistically significant after correcting for multiple comparisons) were between COX2 and change in osteophyte grade (P < 0.001) and Kellgren-Lawrence grade (P < 2 x 10( 5)), and between the genes for vitamin D receptor (P < 0.002) and a thrombospondin (THSD2) (P < 0.002) and change in osteophyte grade. CONCLUSIONS: Our results suggest a role for genes regulating inflammatory pathways in the radiographic progression of spine degeneration. This could prove a fruitful area for future therapeutics for the spine and other joints. PMID- 16261125 TI - Anomalous vertebral artery at the extraosseous and intraosseous regions of the craniovertebral junction: analysis by three-dimensional computed tomography angiography. AB - STUDY DESIGN: This study examined the extraosseous and intraosseous anomalies of vertebral arteries in patients who underwent surgery of the craniovertebral junction. OBJECTIVES: To describe the usefulness of three-dimensional computed tomography angiography for evaluating vertebral artery anomalies before surgery. SUMMARY OF BACKGROUND DATA: Previous studies using catheter angiograms have identified anomalous courses of the vertebral artery at the craniovertebral junction. Studies using computed tomography reconstruction also showed deviation of the vertebral artery groove at the C2 isthmus, demonstrating a risk of vertebral artery injury for C1-C2 transarticular screw placement. These analyses provided us with useful information for identifying anomalies of the vertebral artery, but they could not visualize the artery and its circumferential osseous tissue simultaneously, nor could they analyze the reciprocal anatomy of both tissues. METHODS: Thirty-one consecutive patients who submitted to surgery at the craniovertebral junction were evaluated before surgery by three-dimensional computed tomography angiography. Eleven of the patients had congenital osseous anomalies at the craniovertebral junction including os odontoideum and ossiculum terminale. Anomalous vertebral arteries at the extraosseous region were visualized by three-dimensional reconstruction images, and the intraosseous deviation of the vertebral artery at the C2 isthmus was evaluated by multiplanar reconstruction images. RESULTS: Extraosseous and/or intraosseous vertebral artery anomalies were detected in 9 cases. Eight of the 9 cases had osseous anomalies at the craniovertebral junction. Abnormal courses of the vertebral artery at the extraosseous region were detected in 4 cases: 2 had fenestration and 2 had persistent first intersegmental artery. Asymmetry of bilateral vertebral arteries was found in 5 cases: the right was dominant in 3 cases and the left in 2 cases. A high-riding vertebral artery at the C2 isthmus was detected in 5 cases. Based on these findings, we modified our surgical approach and the screw placement; consequently, no vertebral artery injury occurred. CONCLUSIONS: In patients having osseous anomalies at the craniovertebral junction, the frequency of vertebral artery anomalies at the extraosseous and intraosseous regions is increased. With preoperative three-dimensional computed tomography angiography, we can precisely identify the anomalous vertebral artery and reduce the risk of intraoperative injury to the vertebral artery, in advance. PMID- 16261126 TI - The prognosis of conservative treatments for lumbar spinal stenosis: analysis of patients over 70 years of age. AB - STUDY DESIGN: A prospective study. OBJECTIVES: To identify outcomes of aged patients with lumbar spinal stenosis (LSS) treated conservatively and to examine factors that control the prognosis. SUMMARY AND BACKGROUND DATA: There have been no reports evaluating the outcomes of conservative treatments for elderly LSS patients. METHODS: A total of 89 patients, 70 years of age and older, who underwent in-hospital conservative treatment were included. The Japanese Orthopedic Association's score (JOA score) and the disturbance level of activities of daily living (ADL) were used for evaluation. Nerve involvement was classified into radicular, cauda equina, and mixed type. Myelographic findings were classified into central defect with or without block and root defect. Associations between disturbance level of ADL, nerve involvement, and myelographic classifications were investigated. RESULTS: The mean JOA score increased from 11.1 points at admission to 15.9 points at discharge, with 14.3 points maintained at the follow-up; 48.8% of radicular type showed no obstacle in ADL at the follow-up compared with 33.3% of the other types; 13.3% of central defect with block showed no obstacle in ADL compared with 47.8% of the other types with significant difference. CONCLUSION: The prognosis of conservative treatment for aged LSS was relatively good. Radicular type may be a candidate for conservative treatment. However, patients with complete block in the myelogram may not respond favorably to conservative treatment. PMID- 16261127 TI - Reliability and validity of adapted Turkish Version of Scoliosis Research Society 22 (SRS-22) questionnaire. AB - STUDY DESIGN: Outcome study to determine the internal consistency, and validity of adapted Turkish version of Scoliosis Research Society-22 (SRS-22) Instrument. OBJECTIVES: To evaluate the validity and reliability of adapted Turkish Version of SRS-22 questionnaire. SUMMARY OF BACKGROUND DATA: The SRS-22 questionnaire is a widely accepted questionnaire to assess the health-related quality of life for scoliotic patients in the United States. However, its adaptation in languages other than the source language is necessary for its multinational use. METHODS: Translation/retranslation of the English version of the SRS-22 was done, and all steps for cross-cultural adaptation process were performed properly by an expert committee. Later, SRS-22 questionnaires and previously validated Short Form-36 (SF-36) outcome instruments were mailed to 82 patients who had been surgically treated for idiopathic scoliosis. All patients had a minimum of 2 years follow up. Fifty-four patients (66%) responded to the first set of questionnaires. Forty seven of the first time respondents returned their second survey. The average age of the 47 patients (12 male, 35 female) was 19.8 years (range, 14-31 years). The two measures of reliability as internal consistency and reproducibility were determined by Cronbach alpha statistics and intraclass correlation coefficient, respectively. Concurrent validity was measured by comparing with an already validated questionnaire (SF-36). Measurement was made using the Pearson correlation coefficient (r). RESULTS: The study demonstrated satisfactory internal consistency with high Cronbach alpha values for the four of the corresponding domains (pain, 0.72; self-image, 0.80; mental health, 0.72; and satisfaction, 0.83). However, the Cronbach alpha value for function/activity domain (0.48) was considerably lower than the original questionnaire. The intraclass correlation coefficient for the same domains was 0.80, 0.82, 0.78, 0.81, and 0.76, respectively, demonstrating a satisfactory test/retest reproducibility. Considering concurrent validity, two domains had excellent correlation (r = 0.75-1), while 9 had good correlation (r = 0.50 to 0.75), and 6 had moderate correlation (r =0.25-0.50). Based on these results, question 18 in the function/activity domain with lower Cronbach alpha value was revised while question 15 was excluded. The revised SRS-22 was given to 30 adolescent idiopathic scoliosis patients not included in the index study. The revision could improve the Cronbach alpha value for function/activity domain from 0.48 to 0.81. CONCLUSION: This study demonstrated that, if measures are to be used across cultures, the items must not only be translated well linguistically but also must be culturally adapted to maintain the content validity of the instrument at a conceptual level across different cultures. This may necessitate several validation studies to ensure and improve consistency in the content and face validity between source and target versions of a questionnaire due to difficulty in detecting subtle differences in the living habits of different cultures. PMID- 16261128 TI - Rates of lumbar disc surgery before and after implementation of multidisciplinary nonsurgical spine clinics. AB - STUDY DESIGN: Correlation study. OBJECTIVES: To assess the rates of lumbar disc surgery in North Jutland County, Denmark, before and after implementation of two nonsurgical spine clinics, and to compare the observed rates with those for the rest of Denmark in the same time periods. SUMMARY OF BACKGROUND DATA: Few studies have addressed initiatives to reduce high rates of lumbar disc surgery by improving nonsurgical care offered to patients with sciatica and low back pain. METHODS: The study was conducted in North Jutland County, Denmark with 500,000 inhabitants (10% of the Danish population). In 1997, two nonsurgical spine clinics were established, along with an educational program for general practitioners. The clinics targeted patients with sciatica of 1 to 3 months' duration, with or without low back pain. Data on rates of lumbar disc surgery were obtained from the National Registry of Patients. RESULTS: The annual rate of lumbar disc operations for patients in North Jutland County decreased from approximately 60 to 80 per 100,000 before 1997 to 40 per 100,000 in 2001 (P = 0.00), and the rate of elective, first-time disc surgeries decreased by approximately two thirds (P = 0.00). In contrast, the annual rate of lumbar disc operations for patients in the rest of Denmark remained unchanged during the same period. CONCLUSIONS: The implementation of multidisciplinary, nonsurgical spine clinics coincided closely with a significant reduction in the rate of lumbar disc surgery. The observed reduction seems most likely to be causally associated with educational activities and improved patient care provided by the clinics. PMID- 16261130 TI - Abnormal motion in spondylolytic spondylolisthesis. PMID- 16261131 TI - Public health strategies for preventing and controlling overweight and obesity in school and worksite settings: a report on recommendations of the Task Force on Community Preventive Services. AB - Reducing morbidity and mortality related to overweight and obesity is a public health priority. Various interventions in school and worksite settings aim to maintain or achieve healthy weight. To identify effective strategies for weight control that can be implemented in these settings, the Task Force on Community Preventive Services (Task Force) has conducted systematic reviews of the evidence on nutrition, physical activity, combinations of these interventions, and other behavioral interventions (e.g., cognitive techniques such as self-awareness and cue recognition). Task Force recommendations are based on evidence of effectiveness, which is defined in this report as achieving a mean weight loss of > or =4 pounds, measured > or =6 months after initiation of the intervention program. The Task Force recommends multicomponent interventions that include nutrition and physical activity (including strategies such as providing nutrition education or dietary prescription, physical activity prescription or group activity, and behavioral skills development and training) to control overweight and obesity among adults in worksite settings. The Task Force determined that insufficient evidence existed to determine the effectiveness of combination nutrition and physical activity interventions to prevent or reduce overweight and obesity in school settings because of the limited number of qualifying studies reporting noncomparable outcomes. This report describes the methods used in these systematic reviews; provides additional information regarding these recommendations; and cites sources for full reviews containing details regarding applicability, other benefits and harms, barriers to implementation, research gaps, and economic data (when available) regarding interventions. PMID- 16261133 TI - Deficits in sexual and aggressive behaviors in Cnga2 mutant mice. AB - Odors detected by the vomeronasal organ or the main olfactory epithelium (MOE) trigger social behaviors in many animals. It is unknown whether MOE neurons detect cues that initiate mating or aggression. We demonstrate that mice lacking functional CNGA2 (cyclic nucleotide-gated channel alpha2), which is required for odor-evoked MOE signaling, fail to mate or fight, suggesting a broad and essential role for the MOE in regulating these behaviors. PMID- 16261132 TI - Synaptic background activity controls spike transfer from thalamus to cortex. AB - Characterizing the responsiveness of thalamic neurons is crucial to understanding the flow of sensory information. Typically, thalamocortical neurons possess two distinct firing modes. At depolarized membrane potentials, thalamic cells fire single action potentials and faithfully relay synaptic inputs to the cortex. At hyperpolarized potentials, the activation of T-type calcium channels promotes burst firing, and the transfer is less accurate. Our results suggest that this duality no longer holds if synaptic background activity is taken into account. By injecting stochastic conductances into guinea-pig thalamocortical neurons in slices, we show that the transfer function of these neurons is strongly influenced by conductance noise. The combination of synaptic noise with intrinsic properties gives a global responsiveness that is more linear, mixing single-spike and burst responses at all membrane potentials. Because in thalamic neurons, background synaptic input originates mainly from cortex, these results support a determinant role of corticothalamic feedback during sensory information processing. PMID- 16261134 TI - BK channel beta4 subunit reduces dentate gyrus excitability and protects against temporal lobe seizures. AB - Synaptic inhibition within the hippocampus dentate gyrus serves a 'low-pass filtering' function that protects against hyperexcitability that leads to temporal lobe seizures. Here we demonstrate that calcium-activated potassium (BK) channel accessory beta4 subunits serve as key regulators of intrinsic firing properties that contribute to the low-pass filtering function of dentate granule cells. Notably, a critical beta4 subunit function is to preclude BK channels from contributing to membrane repolarization and thereby broaden action potentials. Longer-duration action potentials secondarily recruit SK channels, leading to greater spike frequency adaptation and reduced firing rates. In contrast, granule cells from beta4 knockout mice show a gain-of-function for BK channels that sharpens action potentials and supports higher firing rates. Consistent with breakdown of the dentate filter, beta4 knockouts show distinctive seizures emanating from the temporal cortex, demonstrating a unique nonsynaptic mechanism for gate control of hippocampal synchronization leading to temporal lobe epilepsy. PMID- 16261135 TI - Dissociation between physical and mental number line bisection in right hemisphere brain damage. AB - To compare numeric quantities, humans make use of a 'mental number line' with smaller quantities located to the left of larger ones; it is unclear, however, whether orienting along the number line is like orienting along a physical line. We found that in brain-damaged subjects with defective leftward orienting, rightward deviation in the bisection of numeric and physical intervals is doubly dissociated. Deviation in numeric interval bisection was associated with prefrontal damage and spatial working memory deficit. PMID- 16261136 TI - Matching storage and recall: hippocampal spike timing-dependent plasticity and phase response curves. AB - Hippocampal area CA3 is widely considered to function as an autoassociative memory. However, it is insufficiently understood how it does so. In particular, the extensive experimental evidence for the importance of carefully regulated spiking times poses the question as to how spike timing-based dynamics may support memory functions. Here, we develop a normative theory of autoassociative memory encompassing such network dynamics. Our theory specifies the way that the synaptic plasticity rule of a memory constrains the form of neuronal interactions that will retrieve memories optimally. If memories are stored by spike timing dependent plasticity, neuronal interactions should be formalized in terms of a phase response curve, indicating the effect of presynaptic spikes on the timing of postsynaptic spikes. We show through simulation that such memories are competent analog autoassociators and demonstrate directly that the attributes of phase response curves of CA3 pyramidal cells recorded in vitro qualitatively conform with the theory. PMID- 16261144 TI - Postop monitoring: on the record? Compromise is in order. PMID- 16261145 TI - Postop monitoring: on the record? PMID- 16261146 TI - Postop monitoring: on the record? Proof is in the records. PMID- 16261147 TI - Postop monitoring: on the record? Consider analgesia for all. PMID- 16261150 TI - Protocol audits for post-approval monitoring of animal use protocols. AB - Monitoring of animal use protocols following IACUC approval is a way to ensure regulatory compliance and to improve communication between the IACUC and investigators. The authors describe the protocol audit system used by the University of California, San Diego's Animal Welfare Program to ensure that IACUC approved protocols are followed. This program, as well as the protocol audit form they provide, can be used by institutions to initiate, enhance, or evaluate post approval monitoring programs. PMID- 16261148 TI - Failure to observe fluoroscopic contrast agent in mouse hind limb. PMID- 16261151 TI - Engaging the IACUC through comprehensive training. AB - The IACUC is one of the most important committees at a research institution and plays a critical role in the success of an animal care and use program. It is the responsibility of the institution to provide IACUC members with adequate and appropriate training. The authors explore various IACUC training options. PMID- 16261152 TI - Defining the Animal Care and Use Program. AB - An effective Animal Care and Use Program is critical to an institution's ability to ensure that animal research is conducted humanely and follows all applicable regulations and guidelines; however, no straightforward definition of the fundamentals of such a Program now exists. The author provides a global view of the key programmatic components, which can be used to improve existing programs or implement new programs. PMID- 16261153 TI - A review of tribromoethanol anesthesia for production of genetically engineered mice and rats. AB - Tribromoethanol (TBE) is easy and inexpensive to make in the laboratory from readily available reagents, requires no special equipment for its administration, and is not subject to federal or state drug enforcement agency regulations. Intraperitoneal (i.p.) injection of TBE results in the simple and rapid induction of short-term surgical anesthesia; however, recent adverse reports about the efficacy and safety of TBE make its continued routine use as a rodent anesthetic controversial. The authors review the history and use of TBE as an animal anesthetic and conclude that TBE should be relegated to acute terminal studies when administered i.p. PMID- 16261155 TI - Head and neck squamous cell carcinoma transcriptome analysis by comprehensive validated differential display. AB - Head and neck squamous cell carcinoma (HNSCC) is common worldwide and is associated with a poor rate of survival. Identification of new markers and therapeutic targets, and understanding the complex transformation process, will require a comprehensive description of genome expression, that can only be achieved by combining different methodologies. We report here the HNSCC transcriptome that was determined by exhaustive differential display (DD) analysis coupled with validation by different methods on the same patient samples. The resulting 820 nonredundant sequences were analysed by high throughput bioinformatics analysis. Human proteins were identified for 73% (596) of the DD sequences. A large proportion (>50%) of the remaining unassigned sequences match ESTs (expressed sequence tags) from human tumours. For the functionally annotated proteins, there is significant enrichment for relevant biological processes, including cell motility, protein biosynthesis, stress and immune responses, cell death, cell cycle, cell proliferation and/or maintenance and transport. Three of the novel proteins (TMEM16A, PHLDB2 and ARHGAP21) were analysed further to show that they have the potential to be developed as therapeutic targets. PMID- 16261156 TI - Chromosomal instability at a mutational hotspot in polyoma middle T-antigen affects its ability to activate the ARF-p53 tumor suppressor pathway. AB - We have isolated spontaneous mutants of polyoma virus middle T-antigen (PyMT) that do not activate the ARF-p53 pathway based on their inability to block REF52 cell division. The REF52 cells containing these mutants have a flat untransformed morphological phenotype and do not express the ARF protein. The PyMT mutations in the different cell isolates so far analysed occur at a mutational hotspot in the PyMT sequence between nucleotides 1241 and 1249, which contains nine consecutive cytosines. In one set of mutants a single cytosine was deleted, while in another mutant set an additional cytosine was inserted. Both these mutations result in frameshifts, generating altered PyMT proteins containing amino-acid sequences derived from each of the two other alternative reading frames of the polyoma virus early region. Both types of mutations result in the loss of the C-terminal PyMT region containing the membrane-binding hydrophobic region and result is mislocalization of the PyMT mutant proteins. Revertant wild-type PyMT (containing nine cytosines) was easily detected in transformants generated after infection of REF52 cells expressing high amounts of dominant negative p53 with retroviruses containing either mutation. We demonstrate that wild-type PyMT revertants are derived from mutations in the hotspot sequence of the integrated mutant PyMT sequences. PMID- 16261158 TI - Multiple alternate p21 transcripts are regulated by p53 in human cells. AB - The p53 tumor-suppressor is a transcription factor that is stabilized in response to cellular stress leading to growth arrest or apoptosis. p21(WAF1/CIP1) is a major transcriptional target of p53 and it plays a critical role in p53-dependent cell cycle arrest. In this study, we identified multiple alternate human p21 transcripts that have their transcriptional start sites in the direct proximity of the distal p53 response element. These transcripts are upregulated as a result of DNA damage-induced p53 activation. Furthermore, the basal expression of these alternate transcripts is strongly regulated by p53 and they are undetectable in p53-knocked down cells. This is in contrast to classical p21 transcripts, which have reduced, albeit detectable expression levels in the absence of p53. The existence of the alternate transcripts underscores the complexity of the human p21 genomic locus and opens up new avenues for further investigation. PMID- 16261157 TI - Expression of cathepsin L in human tumor cells is under the control of distinct regulatory mechanisms. AB - Cathepsin L, a cysteine protease, is overexpressed in human tumor cells and plays a major role in melanoma progression. Our aim was herein to identify molecular mechanisms, which contribute to its overexpression. We found that cathepsin L protein expression correlated with mRNA level in tumor cells. Therefore, we focused on mechanisms involved in cathepsin L mRNA regulation. CpG island was localized in the 5' region of cathepsin L gene that encompassed regulatory regions identified as essential for promoter activity. CpG dinucleotides, not methylated in any melanoma cells analysed, were methylated in a B lymphoma cell line, which poorly express cathepsin L. Our data demonstrate that in lymphoma cells, cathepsin L silencing was methylation-dependent. Furthermore, gene amplification was involved in cathepsin L overexpression in one melanoma cell line, while transcriptional mechanisms but not mRNA stability are responsible of cathepsin L overexpression in others melanoma cells. In addition, NF-Y, Sp1, Sp2 and Sp3 transcription factors, essential to basal cathepsin L transcription, are not directly involved in overexpression. Thus, our data provides the first demonstration that cathepsin L expression in tumor cells is under the control of distinct molecular mechanisms. PMID- 16261160 TI - HTPAP gene on chromosome 8p is a candidate metastasis suppressor for human hepatocellular carcinoma. AB - Our previous studies suggested that chromosome 8p deletion is associated with metastasis of hepatocellular carcinoma (HCC), in which some novel metastasis suppressor genes might be harbored. The present study aimed to identify the metastatic suppressor gene(s). A cDNA chip was constructed with the expressed sequence tags (ESTs) from chromosome 8p and used to compare the difference of expression profiling between the MHCC97-H and MHCC97-L cell lines with different metastatic potentials and similar genetic backgrounds. In all, 10 ESTs were significantly downregulated in MHCC97-H cell line with higher metastatic potential. One full-length gene, HTPAP (phosphatidic acid phosphatase type 2 domain containing 1B), was identified at chromosome 8p12. Sequencing and bioinformatic analyses revealed that HTPAP has 826 bp and encodes a putative protein of 175 amino acids with a transmembrane segment at the NH2 terminus, two protein kinase C phosphorylation site and one tyrosine kinase phosphorylation site. Its expression level in metastatic tumor tissues was much lower than that of primary HCC tissues. Both in vitro and in vivo assays suggested that HTPAP could suppress the invasion and metastasis of HCC. These suggested that HTPAP is a novel metastatic suppressor gene for HCC. The mechanism of the effect of HTPAP on HCC metastasis is not clear yet and deserves further investigation. PMID- 16261161 TI - GPI-anchored TIMP-1 treatment renders renal cell carcinoma sensitive to FAS meditated killing. AB - The resistance of tumours to immune-mediated lysis has been linked to the biology of matrix metalloproteinases (MMPs), and specifically to the cell surface expression of MMPs by the tumour cell. The endogenous tissue inhibitors of metalloproteinases (TIMPs) exhibit diverse physiological/biological functions including the moderation of tumour growth, metastasis and apoptosis. These biologic activities are mediated in part by the stoichiometry of TIMP/MMP/cell surface protein interactions. A glycosylphosphatidylinositol (GPI) anchor was fused to TIMP-1 to focus defined concentrations of this inhibitory protein on the surface of three renal cell carcinoma (RCC) cell lines (RCC-26, RCC-53 and A498) independently of cell surface protein-protein interactions. Exogenously added TIMP-1-GPI efficiently inserted into the RCC cell membrane and dramatically altered the association of MMPs with the cell surface. TIMP-1-GPI treatment inhibited RCC proliferation and rendered the normally FAS-resistant RCC cells sensitive to FAS-induced apoptosis but did not alter perforin-mediated lysis by cytotoxic effector cells. The increased sensitivity to FAS-mediated apoptosis correlated with an alteration in the balance of pro- and antiapoptotic BCL-2 family proteins. By interfering with the proliferative capacity and inducing sensitivity to immune effector mechanisms GPI-anchored TIMP-1 may represent a more effective version of the TIMP-1 protein for therapeutic strategies. PMID- 16261159 TI - Cell adhesion and prostate tumor-suppressor activity of TSLL2/IGSF4C, an immunoglobulin superfamily molecule homologous to TSLC1/IGSF4. AB - The TSLL2/IGSF4C encodes an immunoglobulin (Ig) superfamily molecule showing significant homology with a lung tumor suppressor, TSLC1. The TSLL2 protein of 55 kDa is mainly expressed in the kidney, bladder, and prostate in addition to the brain. Here, we report the biological significance of TSLL2 in the urinary tissues. An immunohistochemical study reveals that TSLL2 is expressed at the cell cell attachment sites in the renal tubules, the transitional epithelia of the bladder, and the glandular epithelia of the prostate. Confocal microscopy analysis demonstrates that TSLL2 is localized in the lateral membranes in polarized Mardin-Darby canine kidney (MDCK) cells. TSLL2 forms homo-dimers and its overexpression induces aggregation of suspended MDCK cells in a Ca2+/Mg2+ independent manner, suggesting that it is involved in cell adhesion through homophilic trans-interaction. The TSLL2 gene is mapped on the chromosomal region 19q13.2, whose loss of heterozygosity has been frequently reported in prostate cancer. TSLL2 protein is lost in nine of nine primary prostate cancers and in a prostate cancer cell, PPC-1. Introduction of TSLL2 into PPC-1 strongly suppresses subcutaneous tumor formation in nude mice. These results suggest that TSLL2 is a new member of the Ig superfamily cell adhesion molecules and is a tumor suppressor candidate in prostate cancer. PMID- 16261164 TI - Laser microdissection and microarray analysis of breast tumors reveal ER-alpha related genes and pathways. AB - About 70-80% of breast cancers express estrogen receptor alpha (ER-alpha), and estrogens play important roles in the development and growth of hormone-dependent tumors. Together with lymph node metastasis, tumor size, and histological grade, ER status is considered as one of the prognostic factors in breast cancer, and an indicator for hormonal treatment. To investigate genes and pathways that are associated with ER status and epithelial cells in breast tumor, we applied laser capture microdissection (LCM) technology to capture epithelial tumor cells from 28 lymph node-negative breast tumor samples, in which 17 patients had ER-alpha+ tumors, and 11 patients have ER-alpha- tumors. Gene expression profiles were analysed on Affymetrix Hu133A GeneChip. Meanwhile, gene profiles using total RNA isolated from bulk tumors of the same 28 patients were also generated. In total, 146 genes and 112 genes with significant P-value and having significant differential expression between ER-alpha+ and ER-alpha- tumors were identified from the LCM data set and bulk tissue data set, respectively. A total of 61 genes were found to be common in both data sets, while 85 genes were unique to the LCM data set and 51 genes were present only in the bulk tumor data set. Pathway analysis with the 85 genes using Gene Ontology suggested that genes involved in endocytosis, ceramide generation, Ras/ERK/Ark cascade, and JAT-STAT pathways may play roles related to ER. The gene profiling with LCM-captured tumor cells provides a unique approach to study epithelial tumor cells and to gain an insight into signaling pathways associated with ER. PMID- 16261163 TI - RAR-mediated epigenetic control of the cytochrome P450 Cyp26a1 in embryocarcinoma cells. AB - Retinoic acid (RA) is a signaling molecule that plays a pivotal role in major cellular processes and vertebrate development. RA action is mediated by specialized transcription factors, the nuclear RA receptors (RARs), which regulate the transcription of genes containing a RA-responsive element (RARE). Here we demonstrate that the genes for the RA-receptor RARbeta2 and the cytochrome P450 RA-specific hydrolase Cyp26a1 involved in RA catabolism are coordinately regulated by RA. We found that both RARbeta2 and Cyp26a1 genes are epigenetically silenced in the absence of DNA methylation in RAC65, a P19 embryocarcinoma cell line derivative carrying a dominant-negative RARalpha mutant and resistant to the growth-inhibitory and differentiation effects of RA. In response to RA, RARbeta2 transcription is epigenetically regulated by RARalpha. Similarly, we found that Cyp26a1 transcription is epigenetically regulated by RARbeta2. Knocking down RARbeta2 transcription by RNA interference in wild-type P19 cells, with an intact RARalpha, induced Cyp26a1 transcriptional repression in the absence of DNA methylation. Concomitantly, cells developed RA resistance and did not undergo RA-induced neuron differentiation. Apparently, RARalpha, RARbeta2 and Cyp26a1 are components of a RA-regulated gene network. Factors affecting an upstream gene of the network can trigger repressive chromatin changes -- which are propagated in a domino fashion - at downstream genes of the network. This study also shows that chromatin inactivity, and consequent transcriptional silencing, can be achieved in the absence of DNA methylation. PMID- 16261162 TI - RelB regulates manganese superoxide dismutase gene and resistance to ionizing radiation of prostate cancer cells. AB - The relationship between NF-kappaB and resistance to radiation treatment in many tumor cell types has been generally well recognized. However, which members of the NF-kappaB family contribute to radiation resistance is unclear. In the present study, we demonstrate that RelB plays an important radioprotective role in aggressive prostate cancer cells, in part by the induction of antioxidant and antiapoptotic manganese superoxide dismutase (MnSOD) gene. RelB is both constitutively present and is inducible by radiation in aggressive prostate cancer cells. Using ectopically expressed dominant negative inhibitor, p100 mutant, and the siRNA approach, we demonstrate that selective inhibition of RelB significantly decreases the levels of MnSOD resulting in a significant increase in the sensitivity of prostate cancer cells to radiation treatment. These results demonstrate that RelB plays an important role in redox regulation of the cell and protects aggressive prostate cancer cells against radiation-induced cell death. Thus, inhibition of RelB could be a novel mechanism to radiosensitize prostate cancer. PMID- 16261165 TI - Renal cell carcinoma risk in type 2 von Hippel-Lindau disease correlates with defects in pVHL stability and HIF-1alpha interactions. AB - The von Hippel-Lindau (VHL) tumor suppressor protein is the substrate binding subunit of the CBC(VHL) E3 ubiquitin ligase complex. Mutations in the VHL gene cause a variety of tumors with complex genotype/phenotype correlations. Type 2A and type 2B VHL disease are characterized by a low or high risk of renal cell carcinoma, respectively. To investigate the molecular basis underlying the difference between disease types 2A and 2B, we performed a detailed biochemical analysis of the two most frequent type 2A mutations, Y98 H and Y112 H, in comparison to type 2B mutations in the same residues, Y98N and Y112N. While none of these mutations affected the assembly of CBC(VHL) complexes, the type 2A mutant proteins exhibited higher stabilities at physiological temperature. Moreover, the type 2A mutant proteins possessed higher binding affinities for the key cellular substrate, hypoxia-inducible transcription factor 1 (HIF-1alpha). Consistent with these results, type 2A but not type 2B mutant VHL proteins retained significant ubiquitin ligase activity towards HIF-1alpha in vitro. We propose that this residual ubiquitin ligase activity is sufficient to suppress renal cell carcinogenesis in vivo. PMID- 16261166 TI - Cdt1 transgenic mice develop lymphoblastic lymphoma in the absence of p53. AB - The exact duplication of chromosomal DNA during each cell cycle ensures the correct inheritance of genetic material from mother to daughter cells. In eukaryotic cells, DNA replication can occur only when the origin of DNA replication is accurately marked by a group of proteins termed licensing proteins. One such protein is Cdt1, which is recruited first to the origin of DNA replication followed by cell division cycle 6 (Cdc6) and mini-chromosome maintenance proteins (Mcms). We previously reported that NIH3T3 cells overexpressing Cdt1 readily formed tumors in mice. To further investigate its oncogenic mechanism, we generated transgenic mice expressing Cdt1 in thymocytes. Our studies demonstrated that T-cell-directed Cdt1 transgenic mice showed normal T-cell development. However, such transgenic mice developed thymic lymphoblastic lymphoma when crossed with p53 null mice. Furthermore, tumor cells derived from NIH3T3 cells overexpressing Cdt1 displayed numerical and structural chromosomal aberrations in the form of ploidy, double minutes, translocation, inversion, chromosome end-to-end fusion and robertsonian mutation. Collectively, our studies suggest that Cdt1 overexpression most likely contributes to tumorigenecity by causing genomic instability. PMID- 16261168 TI - Autism spectrum disorders associated with X chromosome markers in French-Canadian males. AB - It is now well established that genetic factors play an important role in the pathogenesis of autism disorder and converging lines of evidence suggest the implication of the X chromosome. Using a sample of subjects diagnosed with autism spectrum disorders, exclusively composed of males from French-Canadian (FC) origin, we tested markers covering the entire X chromosome using a family-based association study. Our initial analysis revealed the presence of association at two loci: DXS6789 (P=0.026) and DXS8043 (P=0.0101). In a second step, we added support to the association at DXS8043 using additional markers, additional subjects and a haplotype-based analysis (best obtained P-value=0.00001). These results provide support for the existence of a locus on the X chromosome that predisposes the FC to autism spectrum disorders. PMID- 16261167 TI - Identification of additional variants within the human dopamine transporter gene provides further evidence for an association with bipolar disorder in two independent samples. AB - The dopamine transporter (DAT) is the site of action of stimulants, and variations in the human DAT gene (DAT1) have been associated with susceptibility to several psychiatric disorders including attention deficit hyperactivity disorder (ADHD) and bipolar disorder. We have previously reported the association of bipolar disorder to novel SNPs in the 3' end of DAT1. We now report the identification of 20 additional SNPs in DAT1 for a total of 63 variants. We also report evidence for association to bipolar disorder in a second independent sample of families. Eight newly identified SNPs and 14 previously identified SNPs were analyzed in two independent samples of 50 and 70 families each using the transmission disequilibrium test. Two of the eight new SNPs, one in intron 8 and one in intron 13, were found to be moderately associated with bipolar disorder, each in one of the two independent samples. Analysis of haplotypes comprised of all 22 SNPs in sliding windows of five adjacent SNPs revealed an association to the region near introns 7 and 8 in both samples (empirical P-values 0.002 and 0.001, respectively, for the same window). The haplotype block structure observed in the gene in our previous study was confirmed in this sample with greater resolution allowing for discrimination of a third haplotype block in the middle of the gene. Together, these data are consistent with the presence of multiple variants in DAT1 that convey susceptibility to bipolar disorder. PMID- 16261169 TI - Precise hit: adeno-associated virus in gene targeting. AB - Vectors based on the adeno-associated virus (AAV) have attracted much attention as potent gene-delivery vehicles, mainly because of the persistence of this non pathogenic virus in the host cell and its sustainable therapeutic gene expression. However, virus infection can be accompanied by potentially mutagenic random vector integration into the genome. A novel approach to AAV-mediated gene therapy based on gene targeting through homologous recombination allows efficient, high-fidelity, non-mutagenic gene repair in a host cell. PMID- 16261170 TI - The bacterial ribosome as a target for antibiotics. AB - Many clinically useful antibiotics exert their antimicrobial effects by blocking protein synthesis on the bacterial ribosome. The structure of the ribosome has recently been determined by X-ray crystallography, revealing the molecular details of the antibiotic-binding sites. The crystal data explain many earlier biochemical and genetic observations, including how drugs exercise their inhibitory effects, how some drugs in combination enhance or impede each other's binding, and how alterations to ribosomal components confer resistance. The crystal structures also provide insight as to how existing drugs might be derivatized (or novel drugs created) to improve binding and circumvent resistance. PMID- 16261171 TI - Deinococcus radiodurans - the consummate survivor. AB - Relatively little is known about the biochemical basis of the capacity of Deinococcus radiodurans to endure the genetic insult that results from exposure to ionizing radiation and can include hundreds of DNA double-strand breaks. However, recent reports indicate that this species compensates for extensive DNA damage through adaptations that allow cells to avoid the potentially detrimental effects of DNA strand breaks. It seems that D. radiodurans uses mechanisms that limit DNA degradation and that restrict the diffusion of DNA fragments that are produced following irradiation, to preserve genetic integrity. These mechanisms also increase the efficiency of the DNA-repair proteins. PMID- 16261172 TI - Opinion: antibody-based therapies for malaria. AB - Antibodies are multifunctional glycoproteins that are found in blood and tissue fluids, and can protect against malaria by binding and neutralizing malaria parasites and preparing them for destruction by immune cells. Important technical advances mean that it is now possible to synthesize antibodies against important Plasmodium antigens that could be used for therapeutic purposes. These reagents could be designed to act like a drug and kill parasites directly, or could be used in vaccine strategies to protect individuals from infection. In this article, we discuss the possible therapeutic uses of antibodies in the treatment and prevention of malaria. PMID- 16261173 TI - Follicular B helper T cells in antibody responses and autoimmunity. AB - T-cell help for B cells is essential for high-affinity antibody responses and B cell memory. Recently, the identity of a discrete follicular population of T cells that has a crucial role in this process has become clearer. Similar to primed CD4(+) T cells in the tonsils and memory CD4(+) T cells in the peripheral blood, this follicular population of T cells expresses CXC-chemokine receptor 5 (CXCR5). Owing to their distinct homing preferences and helper function, these T cells differ from T helper 1 and T helper 2 cells and have been denoted follicular B helper T cells. Here, we outline the central role of this subset in normal and pathological immune responses. PMID- 16261175 TI - Molecular defects in T- and B-cell primary immunodeficiency diseases. AB - More than 120 inherited primary immunodeficiency diseases have been discovered in the past five decades, and the precise genetic defect in many of these diseases has now been identified. Increasing understanding of these molecular defects has considerably influenced both basic and translational research, and this has extended to many branches of medicine. Recent advances in both diagnosis and therapeutic modalities have allowed these defects to be identified earlier and to be more precisely defined, and they have also resulted in more promising long term outcomes. The prospect of gene therapy continues to be included in the armamentarium of treatment considerations, because these conditions could be among the first to benefit from gene-therapy trials in humans. PMID- 16261176 TI - Twenty-first-century medical microbiology services in the UK. AB - With infection once again a high priority for the UK National Health Service (NHS), the medical microbiology and infection-control services require increased technology resources and more multidisciplinary staff. Clinical care and health protection need a coordinated network of microbiology services working to consistent standards, provided locally by NHS Trusts and supported by the regional expertise and national reference laboratories of the new Health Protection Agency. Here, I outline my thoughts on the need for these new resources and the ways in which clinical microbiology services in the UK can best meet the demands of the twenty-first century. PMID- 16261177 TI - A comparative genomic view of clostridial sporulation and physiology. AB - Clostridia are anaerobic, endospore-forming prokaryotes that include strains of importance to human and animal health and physiology, cellulose degradation, solvent production and bioremediation. Their differentiation and related developmental programmes are not well understood at the molecular level. Recent genome sequencing and transcriptional-profiling studies have offered a glimpse of their inner workings and indicate that a better understanding of the orchestration of the molecular events that underlie their unique physiology, capabilities and diversity will pay major dividends. PMID- 16261174 TI - Reconstructing immune phylogeny: new perspectives. AB - Numerous studies of the mammalian immune system have begun to uncover profound interrelationships, as well as fundamental differences, between the adaptive and innate systems of immune recognition. Coincident with these investigations, the increasing experimental accessibility of non-mammalian jawed vertebrates, jawless vertebrates, protochordates and invertebrates has provided intriguing new information regarding the likely patterns of emergence of immune-related molecules during metazoan phylogeny, as well as the evolution of alternative mechanisms for receptor diversification. Such findings blur traditional distinctions between adaptive and innate immunity and emphasize that, throughout evolution, the immune system has used a remarkably extensive variety of solutions to meet fundamentally similar requirements for host protection. PMID- 16261178 TI - Neuroenergetics and the kinetic design of excitatory synapses. AB - Why is the characteristic timescale of neural information processing in the millisecond range, corresponding to a 'clock speed' of about 1 kHz, whereas the clock speed of modern computers is about 3 GHz? Here we investigate how the brain's energy supply limits the maximum rate at which the brain can compute, and how the molecular components of excitatory synapses have evolved properties that are matched to the information processing they perform. PMID- 16261179 TI - Pathways modulating neural KCNQ/M (Kv7) potassium channels. AB - K(+) channels play a crucial role in regulating the excitability of neurons. Many K(+) channels are, in turn, regulated by neurotransmitters. One of the first neurotransmitter-regulated channels to be identified, some 25 years ago, was the M channel. This was categorized as such because its activity was inhibited through stimulation of muscarinic acetylcholine receptors. M channels are now known to be composed of subunits of the Kv7 (KCNQ) K(+) channel family. However, until recently, the link between the receptors and the channels has remained elusive. Here, we summarize recent developments that have begun to clarify this link and discuss their implications for physiology and medicine. PMID- 16261180 TI - Synaptic plasticity at hippocampal mossy fibre synapses. AB - The dentate gyrus provides the main input to the hippocampus. Information reaches the CA3 region through mossy fibre synapses made by dentate granule cell axons. Synaptic plasticity at the mossy fibre-pyramidal cell synapse is unusual for several reasons, including low basal release probability, pronounced frequency facilitation and a lack of N-methyl-D-aspartate receptor involvement in long-term potentiation. In the past few years, some of the mechanisms underlying the peculiar features of mossy fibre synapses have been elucidated. Here we describe recent work from several laboratories on the various forms of synaptic plasticity at hippocampal mossy fibre synapses. We conclude that these contacts have just begun to reveal their many secrets. PMID- 16261181 TI - Critical period plasticity in local cortical circuits. AB - Neuronal circuits in the brain are shaped by experience during 'critical periods' in early postnatal life. In the primary visual cortex, this activity-dependent development is triggered by the functional maturation of local inhibitory connections and driven by a specific, late-developing subset of interneurons. Ultimately, the structural consolidation of competing sensory inputs is mediated by a proteolytic reorganization of the extracellular matrix that occurs only during the critical period. The reactivation of this process, and subsequent recovery of function in conditions such as amblyopia, can now be studied with realistic circuit models that might generalize across systems. PMID- 16261182 TI - Science and society: death, unconsciousness and the brain. AB - The concept of death has evolved as technology has progressed. This has forced medicine and society to redefine its ancient cardiorespiratory centred diagnosis to a neurocentric diagnosis of death. The apparent consensus about the definition of death has not yet appeased all controversy. Ethical, moral and religious concerns continue to surface and include a prevailing malaise about possible expansions of the definition of death to encompass the vegetative state or about the feared bias of formulating criteria so as to facilitate organ transplantation. PMID- 16261183 TI - Short-term changes in inflammatory response protein (hsCRP) do not parallel with changes in coronary vasoreactivity in obese men. AB - AIM: Obese subjects are characterized by increased high-sensitivity C-reactive protein (hsCRP) and coronary vascular resistance. Clucocorticoids suppress inflammation, a possible cardioprotective effect. We tested the short-term anti inflammatory effect of dexamethasone (dx) on these parameters in obese subjects. METHODS: Coronary vascular resistance was quantitated basally and during adenosine infusion with or without simultaneous euglycemic hyperinsulinemic clamp (insulin infusion rate of 1 mU/kg/min) in 11 obese and 19 age-matched nonobese males using positron emission tomography and (15)O-water. Each subject was studied both with and without previous dx treatment for 2 days (2 mg/day). RESULTS: Before dx treatment, hsCRP concentration was significantly higher in obese than in nonobese subjects (1.55+/-1.73 vs 0.32+/-0.32 mg/l, P = 0.005). In addition, coronary vascular resistances were higher (P < 0.05) in obese than in nonobese subjects at baseline (139+/-36 vs 117+/-22) and during adenosine infusion without (32+/-7 vs 26+/-7) or with simultaneous clamp (26+/-8 vs 21+/-5 mmHg min g/ ml). Dx treatment decreased significantly hsCRP concentration in obese but not in nonobese subjects, leading to similar hsCRP concentrations between the groups (0.45+/-0.43 vs 0.26+/-0.42 mg/l, respectively, P = 0.3). Dx had no effect on coronary vascular resistances (NS). CONCLUSIONS: Obese subjects are characterized by high hsCRP, which can be normalized by dx. However, despite this, coronary vascular resistances did not decrease in obese subjects. Short term changes in inflammatory response protein appear not to parallel with changes in coronary vasoreactivity in obese men. PMID- 16261184 TI - Relationship between severity of nocturnal desaturation and adaptive thermogenesis: preliminary data of apneic patients tested in a whole-body indirect calorimetry chamber. AB - The purpose of this study was to investigate the possibility of a relationship between the severity of obstructive sleep apnea syndrome (OSAS) and adaptive thermogenesis. Daily energy expenditure (DEE) and sleeping metabolic rate (SMR) were measured in apneic and a priori nonapneic subjects who were tested in a whole-body indirect calorimetry chamber for 24 h. The apneic patients were diagnosed by nocturnal home oximetry to determine the percentage of total recording time spent below 90% arterial oxygen saturation (% TRT <90% SaO(2)). Reference equations established from body weight and age in nonapneic subjects were used to predict DEE and SMR in apneic patients. The predicted values of the apneic patients were then compared to their measured values. No significant difference was found between predicted and measured values in SMR nor in DEE. We observed a significant relationship between the severity of nocturnal desaturation and the difference between predicted and measured DEE in apneic patients (r = -0.74, P < 0.05) and a similar negative trend with SMR (r = -0.65, P = 0.08). These preliminary data suggest that a nocturnal hypoxia may influence adaptive thermogenesis in apneic patients and complicate their body weight regulation. PMID- 16261185 TI - The 2nd-4th digit ratio (2D:4D) and neck circumference: implications for risk factors in coronary heart disease. AB - BACKGROUND: The ratio of the lengths of the 2nd and 4th digit (2D:4D) is negatively related to prenatal and adult concentrations of testosterone (T). Testosterone appears to be a protective against myocardial infarction (MI) in men as men with low 2D:4D are older at first MI than men with high 2D:4D, and men with coronary artery disease have lower T levels than men with normal angiograms. Neck circumference (NC), a simple and time-saving screening measure to identify obesity is reported to be positively correlated with the factors of the metabolic syndrome, a complex breakdown of normal physiology characterized by obesity, insulin resistance, hyperlipidemia, and hypertension, and is therefore likely to increase the risk of coronary heart disease (CHD). OBJECTIVE: To investigate possible associations between 2D:4D ratios and NC in men and women. RESEARCH METHODS AND PROCEDURES: 2D:4D ratios, NC, along with measures of waist and hip circumferences, body mass index (BMI), and waist-to-hip ratio was recorded from 127 men and 117 women. RESULTS: A significant positive correlation between 2D:4D and NC and was found for men but not for women after controlling for body mass index (BMI); the higher the ratio the higher the NC. DISCUSSION: This finding supports the suggestion of NC to serve as a predictor for increased risk for CHD as previously suggested. In addition, the present association suggests a predisposition for men towards CHD via 2D:4D as proxy to early sex-steroid exposure. PMID- 16261186 TI - Microsatellite polymorphism of the human leptin gene (LEP) and risk of cardiovascular disease. AB - BACKGROUND: No data have been so far reported on the relationship between polymorphisms of LEP gene and cardiovascular disease. PATIENTS AND METHODS: We genotyped a tetranucleotide repeat mapped in the 3'UTR of the LEP gene (LEP-tet) in 109 subjects with cardiovascular events and in 109 control subjects. RESULTS: Univariate analysis and multivariate logistic regression analysis adjusted for age, gender, smoking status, history of hyperlipidemia, hypertension or diabetes showed not significant association between the genotype of the LEP-tet and cardiovascular diseases. Moreover, no differences were observed in the plasma leptin concentrations between cases and control subjects (22 +/- 19 vs 22 +/- 14 ng/ml, P = 0.52) and in relation to the LEP-tet classes or carriage of specific alleles (P = 0.76 for the association between LEP-tet classes and leptin levels in overall analysis). CONCLUSIONS: In conclusion, our data do not support an association between the LEP-tet microsatellite polymorphism of the human LEP gene and cardiovascular diseases. PMID- 16261187 TI - ALOX5AP expression, but not gene haplotypes, is associated with obesity and insulin resistance. AB - OBJECTIVE: Inflammation in adipose tissue may link obesity to insulin resistance and atherosclerosis. Arachidonate 5-lipoxygenase activating protein (ALOX5AP) gene is involved in the pathogenesis of atherosclerotic cardiovascular disease (CVD). We investigated ALOX5AP expression in adipose tissue, and association of gene polymorphisms with obesity and insulin resistance. DESIGN: For gene expression analysis in adipose tissue, we studied 12 lean and 36 obese women, eight lean and 13 obese men, and nine women before and 2-4 years after gastric banding surgery. For genetic analysis, we studied 231 nonobese and 350 obese men. RESULTS: The ALOX5AP protein, 5-lipoxygenase activating protein (FLAP), as well as 5-lipoxygenase (5LO) itself, were detected in adipocytes. The mRNA expression of ALOX5AP in subcutaneous adipose tissue was increased in obesity and normalized following weight reduction. High adipose tissue mRNA expression of ALOX5AP is associated with insulin resistance as measured by homeostasis model assessment (HOMA(IR)). ALOX5AP haplotypes that associate with CVD are not associated with obesity or insulin resistance. CONCLUSION: ALOX5AP is present in adipose tissue, where its expression is associated with body weight and HOMA(IR), and may provide a link between adipose tissue, inflammation and insulin resistance. Investigated ALOX5AP haplotypes are not major primary risk factors for obesity and insulin resistance. PMID- 16261188 TI - Value of body fat mass vs anthropometric obesity indices in the assessment of metabolic risk factors. AB - OBJECTIVE: To compare the value of body fat mass (%FM) to indirect measures of general (body mass index (BMI)) and central adiposity (waist circumference (WC); waist-to-height ratio (WC/ht)) for the prediction of overweight- and obesity related metabolic risk in a study population with a high prevalence of metabolic syndrome (MSX). METHODS: BMI, WC, WC/ht, body composition (by air-displacement plethysmography) and metabolic risk factors: triglycerides, cholesterol, HDL cholesterol (HDL-C), uric acid, systolic blood pressure (BPsys), insulin resistance by homeostasis model assessment (HOMA-IR) and C-reactive protein (CRP) were measured in 335 adults (191 women, 144 men; mean age 53 +/-13.9 years, prevalence of MSX 30%). RESULTS: When compared with BMI and WC, %FM showed weaker associations with metabolic risk factors, except for CRP and BPsys in men. In women, HDL-C and HOMA-IR showed the closest correlations with BMI. For all other risk factors, WC or WC/ht were the best predictors in both sexes. Differences in the strength of correlations between an obesity index and different risk factors exceeded the differences observed between all obesity indices within one risk factor. In stepwise multiple regression analyses, WC/ht was the main predictor of metabolic risk in both sexes combined. However, analysis of the area under receiver operating characteristic curves for prediction of the prevalence of >or=2 component traits of the MSX revealed a similar accuracy of all obesity indices. CONCLUSIONS: At the population level, measurement of body FM has no advantage over BMI and WC in the prediction of obesity-related metabolic risk. Although measures of central adiposity (WC, WC/ht) tended to show closer associations with risk factors than measures of general adiposity, the differences were small and depended on the type of risk factor and sex, suggesting an equivalent value of methods. PMID- 16261189 TI - The diverse functions of histone lysine methylation. AB - Covalent modifications of histone tails have fundamental roles in chromatin structure and function. One such modification, lysine methylation, has important functions in many biological processes that include heterochromatin formation, X chromosome inactivation and transcriptional regulation. Here, we summarize recent advances in our understanding of how lysine methylation functions in these diverse biological processes, and raise questions that need to be addressed in the future. PMID- 16261190 TI - Growth of the plant cell wall. AB - Plant cells encase themselves within a complex polysaccharide wall, which constitutes the raw material that is used to manufacture textiles, paper, lumber, films, thickeners and other products. The plant cell wall is also the primary source of cellulose, the most abundant and useful biopolymer on the Earth. The cell wall not only strengthens the plant body, but also has key roles in plant growth, cell differentiation, intercellular communication, water movement and defence. Recent discoveries have uncovered how plant cells synthesize wall polysaccharides, assemble them into a strong fibrous network and regulate wall expansion during cell growth. PMID- 16261191 TI - Flux balance analysis of mycolic acid pathway: targets for anti-tubercular drugs. AB - Mycobacterium tuberculosis is the focus of several investigations for design of newer drugs, as tuberculosis remains a major epidemic despite the availability of several drugs and a vaccine. Mycobacteria owe many of their unique qualities to mycolic acids, which are known to be important for their growth, survival, and pathogenicity. Mycolic acid biosynthesis has therefore been the focus of a number of biochemical and genetic studies. It also turns out to be the pathway inhibited by front-line anti-tubercular drugs such as isoniazid and ethionamide. Recent years have seen the emergence of systems-based methodologies that can be used to study microbial metabolism. Here, we seek to apply insights from flux balance analyses of the mycolic acid pathway (MAP) for the identification of anti tubercular drug targets. We present a comprehensive model of mycolic acid synthesis in the pathogen M. tuberculosis involving 197 metabolites participating in 219 reactions catalysed by 28 proteins. Flux balance analysis (FBA) has been performed on the MAP model, which has provided insights into the metabolic capabilities of the pathway. In silico systematic gene deletions and inhibition of InhA by isoniazid, studied here, provide clues about proteins essential for the pathway and hence lead to a rational identification of possible drug targets. Feasibility studies using sequence analysis of the M. tuberculosis H37Rv and human proteomes indicate that, apart from the known InhA, potential targets for anti-tubercular drug design are AccD3, Fas, FabH, Pks13, DesA1/2, and DesA3. Proteins identified as essential by FBA correlate well with those previously identified experimentally through transposon site hybridisation mutagenesis. This study demonstrates the application of FBA for rational identification of potential anti-tubercular drug targets, which can indeed be a general strategy in drug design. The targets, chosen based on the critical points in the pathway, form a ready shortlist for experimental testing. PMID- 16261194 TI - SNPdetector: a software tool for sensitive and accurate SNP detection. AB - Identification of single nucleotide polymorphisms (SNPs) and mutations is important for the discovery of genetic predisposition to complex diseases. PCR resequencing is the method of choice for de novo SNP discovery. However, manual curation of putative SNPs has been a major bottleneck in the application of this method to high-throughput screening. Therefore it is critical to develop a more sensitive and accurate computational method for automated SNP detection. We developed a software tool, SNPdetector, for automated identification of SNPs and mutations in fluorescence-based resequencing reads. SNPdetector was designed to model the process of human visual inspection and has a very low false positive and false negative rate. We demonstrate the superior performance of SNPdetector in SNP and mutation analysis by comparing its results with those derived by human inspection, PolyPhred (a popular SNP detection tool), and independent genotype assays in three large-scale investigations. The first study identified and validated inter- and intra-subspecies variations in 4,650 traces of 25 inbred mouse strains that belong to either the Mus musculus species or the M. spretus species. Unexpected heterozygosity in CAST/Ei strain was observed in two out of 1,167 mouse SNPs. The second study identified 11,241 candidate SNPs in five ENCODE regions of the human genome covering 2.5 Mb of genomic sequence. Approximately 50% of the candidate SNPs were selected for experimental genotyping; the validation rate exceeded 95%. The third study detected ENU induced mutations (at 0.04% allele frequency) in 64,896 traces of 1,236 zebra fish. Our analysis of three large and diverse test datasets demonstrated that SNPdetector is an effective tool for genome-scale research and for large-sample clinical studies. SNPdetector runs on Unix/Linux platform and is available publicly (http://lpg.nci.nih.gov). PMID- 16261192 TI - Genomic variability within an organism exposes its cell lineage tree. AB - What is the lineage relation among the cells of an organism? The answer is sought by developmental biology, immunology, stem cell research, brain research, and cancer research, yet complete cell lineage trees have been reconstructed only for simple organisms such as Caenorhabditis elegans. We discovered that somatic mutations accumulated during normal development of a higher organism implicitly encode its entire cell lineage tree with very high precision. Our mathematical analysis of known mutation rates in microsatellites (MSs) shows that the entire cell lineage tree of a human embryo, or a mouse, in which no cell is a descendent of more than 40 divisions, can be reconstructed from information on somatic MS mutations alone with no errors, with probability greater than 99.95%. Analyzing all approximately 1.5 million MSs of each cell of an organism may not be practical at present, but we also show that in a genetically unstable organism, analyzing only a few hundred MSs may suffice to reconstruct portions of its cell lineage tree. We demonstrate the utility of the approach by reconstructing cell lineage trees from DNA samples of a human cell line displaying MS instability. Our discovery and its associated procedure, which we have automated, may point the way to a future "Human Cell Lineage Project" that would aim to resolve fundamental open questions in biology and medicine by reconstructing ever larger portions of the human cell lineage tree. PMID- 16261195 TI - Ultrasensitization: switch-like regulation of cellular signaling by transcriptional induction. AB - Cellular signaling networks are subject to transcriptional and proteolytic regulation under both physiological and pathological conditions. For example, the expression of proteins subject to covalent modification by phosphorylation is known to be altered upon cellular differentiation or during carcinogenesis. However, it is unclear how moderate alterations in protein expression can bring about large changes in signal transmission as, for example, observed in the case of haploinsufficiency, where halving the expression of signaling proteins abrogates cellular function. By modeling a fundamental motif of signal transduction, the phosphorylation-dephosphorylation cycle, we show that minor alterations in the concentration of the protein subject to phosphorylation (or the phosphatase) can affect signal transmission in a highly ultrasensitive fashion. This "ultrasensitization" is strongly favored by substrate sequestration on the catalyzing enzymes, and can be observed with experimentally measured enzymatic rate constants. Furthermore, we show that coordinated transcription of multiple proteins (i.e., synexpression) within a protein kinase cascade results in even more pronounced all-or-none behavior with respect to signal transmission. Finally, we demonstrate that ultrasensitization can account for specificity and modularity in the regulation of cellular signal transduction. Ultrasensitization can result in all-or-none cell-fate decisions and in highly specific cellular regulation. Additionally, switch-like phenomena such as ultrasensitization are known to contribute to bistability, oscillations, noise reduction, and cellular heterogeneity. PMID- 16261197 TI - Ten simple rules for getting published. PMID- 16261196 TI - Dissimilatory metabolism of nitrogen oxides in bacteria: comparative reconstruction of transcriptional networks. AB - Bacterial response to nitric oxide (NO) is of major importance since NO is an obligatory intermediate of the nitrogen cycle. Transcriptional regulation of the dissimilatory nitric oxides metabolism in bacteria is diverse and involves FNR like transcription factors HcpR, DNR, and NnrR; two-component systems NarXL and NarQP; NO-responsive activator NorR; and nitrite-sensitive repressor NsrR. Using comparative genomics approaches, we predict DNA-binding motifs for these transcriptional factors and describe corresponding regulons in available bacterial genomes. Within the FNR family of regulators, we observed a correlation of two specificity-determining amino acids and contacting bases in corresponding DNA recognition motif. Highly conserved regulon HcpR for the hybrid cluster protein and some other redox enzymes is present in diverse anaerobic bacteria, including Clostridia, Thermotogales, and delta-proteobacteria. NnrR and DNR control denitrification in alpha- and beta-proteobacteria, respectively. Sigma-54 dependent NorR regulon found in some gamma- and beta-proteobacteria contains various enzymes involved in the NO detoxification. Repressor NsrR, which was previously known to control only nitrite reductase operon in Nitrosomonas spp., appears to be the master regulator of the nitric oxides' metabolism, not only in most gamma- and beta-proteobacteria (including well-studied species such as Escherichia coli), but also in gram-positive Bacillus and Streptomyces species. Positional analysis and comparison of regulatory regions of NO detoxification genes allows us to propose the candidate NsrR-binding motif. The most conserved member of the predicted NsrR regulon is the NO-detoxifying flavohemoglobin Hmp. In enterobacteria, the regulon also includes two nitrite-responsive loci, nipAB (hcp-hcr) and nipC (dnrN), thus confirming the identity of the effector, i.e. nitrite. The proposed NsrR regulons in Neisseria and some other species are extended to include denitrification genes. As the result, we demonstrate considerable interconnection between various nitrogen-oxides-responsive regulatory systems for the denitrification and NO detoxification genes and evolutionary plasticity of this transcriptional network. PMID- 16261198 TI - [Preventive and therapeutic effects of recombinant IFN-alpha2b nasal spray on SARS-CoV infection in Macaca mulata]. AB - OBJECTIVE: To study the preventive and therapeutic effects of recombinant IFN alpha2b for nasal spray on SARS-CoV infection in Macaca mulata (rhesus monkey). METHODS: Ten rhesus monkeys were divided into two groups, 5 in interferon group, and 5 in control group. Before and after SARS-CoV attack, the virus was detected in samples such as pharyngeal swab in all the two groups by Real-time PCR (RT PCR) and virus isolation was performed. RESULTS: After virus attack, the level of SARS-CoV-specific IgG and neutralizing antibody were induced by SARS-CoV in the interferon group was weaker than in control group. Hematology items showed no apparent changes after virus attack in treated group. Through pathological examination, the morphology of the lung tissues of two Macaques in the treated group was normal, while the other three displayed the interstitial pneumonia with the thickened septum and infiltration with mononuclear cells. Among which, one monkey showed part of thickened septum fused with each other. These lesions in the interferon treated animals were similar to those seen in the animals in control group, but with smaller scope of pathological changes. No significant abnormity was detected in other organs. CONCLUSION: Recombinant IFN-alpha2b could effectively interdict or weaken SARS-CoV injury in monkeys. PMID- 16261199 TI - [A field trial for evaluating the safety of recombinant human interferon alpha-2b for nasal spray]. AB - OBJECTIVE: To evaluate the safety of recombinant human interferon alpha-2b for nasal spray for the prevention of SARS and other upper respiratory viral infections. METHODS: Field epidemiologic evaluation was conducted, the design was randomized and had a synchronously parallel control group. In the study, the drugs were given for five days and all subjects were followed up for ten days. RESULTS: During the period of using interferon, body temperature of the experimental group was normal compared to the control group. Experimental group had more influenza-like symptoms than the control group (P < 0.05), such as headache (4.83%-7.09%), dizziness (7.17%-11.63%), lassitude (8.55%-15.06%), muscular soreness (4.43%-7.09%), pharynx dryness (12.10%-17.85%), angina (6.25% 8.72%), abdominal pain (2.30%-5.50%) and diarrhea (2.45%-5.66%). Most of side effects reached their peak with in the first 3 days. Except for pharynx dryness, the incidences of all other side effects declined after completion of the use of the trial drug, and incidences of some symptoms in experimental group were lower than those of the control group. There were no significant differences in the symptoms of cough and expectoration between the experimental group and the control group. The incidence of exanthem in the control group was significantly higher than that in the experimental group. The side effect of bloody nasal mucus was not observed in experimental group, which had been reported by other authors in several volunteer studies. CONCLUSION: Using recombinant human interferon alpha-2b for nasal spray could lead to some influenza-like symptoms, however, all those symptoms were mild , reversible, and relieved after completion of the use of the trial drug. No serious side effects were found during the period of following up. The authors conclude that the drug is safe. PMID- 16261200 TI - [A field trial of recombinant human interferon alpha-2b for nasal spray to prevent SARS and other respiratory viral infections]. AB - OBJECTIVE: To study the preventive effect of recombinant human interferon alpha 2b for nasal spray against SARS and other common respiratory viral infections by serum-epidemiological method. METHODS: A randomized, placebo-controlled, double blind field trial study in populations with 14,391 persons from SARS prevalent cities or provinces in China during May-Jun, 2003 and Dec-Apr, 2004. Interferon alpha-2b was given twice per day, once 9 x 10(5) IU by nasal spray for 5 days. Serum samples were taken at 15 days after last administration. Serological tests included SARS IgG antibody and IgM antibodies against influenza B, parainfluenza virus types 1-3, adenovirus type 3, 7 and respiratory syncytial virus by using commercial ELISA kits. RESULTS: No statistically significant difference in serum SARS IgG antibody positive rate was found between the interferon and control groups among 2,757 serum samples. On the other hand, after using interferon, all four respiratory viruses (parainfluenza virus types 1-3 influenza B, adenovirus types 3, 7 and respiratory syncytial virus) in interferon group had lower IgM antibody positive rates than those in control group. Among them there were statistically significant differences between the interferon and control groups for parainfluenza virus, influenza B and adenovirus. The preventive efficacy of interferon against four respiratory viruses was different, from high to low, the rank was Flu B (66.76%), parainfluenza types 1-3 (66.75%), RSV (39.61%) and adenovirus (32.86%). The average preventive efficacy was 50.27%. CONCLUSION: The recombinant human interferon alpha-2b for nasal spray could decrease the rates of common respiratory viruses infection in the selected population. PMID- 16261201 TI - [Preliminary study on nasal spray of interferon alpha-2b used for prevention of rubella and measles virus infections]. AB - OBJECTIVE: To evaluate the efficacy of the interferon alpha-2b nasal spray in prevention of rubella and measles virus infections. METHODS: The properly selected volunteer groups have been divided into interferon alpha-2b experimental and control group. The experimental group received interferon alpha-2b treatment by nasal spray for 2 days before the immunization, then both groups were challenged with rubella and measles attenuated live vaccine respectively through nasal spray. The sera from pre-immunization and 21 and 28 days after immunization were collected to test the IgG antibody titers. The influence on the viral antibody titer reflects the viral preventive effect by interferon alpha-2b. RESULTS: The antibody titer difference of measles virus between experimental and control group was 1.26 (21 day) and 2.96 (28 day), there were statistically difference between them; the difference of rubella virus was 0.95 (21 day) and 0.37 (28 day), but there were no statistically differences found. CONCLUSION: The preliminary results showed that the interferon alpha-2b can be used as prevention method for measles and rubella viral infections. PMID- 16261202 TI - [Cloning, expression and purification of interferon-kappa, a novel human interferon, and its antiviral activity]. AB - OBJECTIVE: To prepare human interferon-k (hIFN-kappa) and study its biological activities. METHODS: Whole length of hIFN-kappa's cDNA was cloned, and its sequence was chemically synthesized according to the optimized codons of E.coli, then was expressed in E.coli DH5alpha. After purified, the rhIFN-kappa protein was tested for its various kinds of biological activities. RESULTS: The purity of rhIFN-kappa was above 90%. In WHIS-VSV system, the antiviral activity of rhIFN kappa was 2.0 x 10(6) IU/mg. Compared with rhIFN-alpha-2b, the biological activities of rhIFN-kappa were all feeble, including antiviral activity, promoting NK cell activity and anti-proliferation activity. CONCLUSION: Antiviral activities of rhIFN-kappa on cell lines of different species are different, different viruses show different sensitivity to rhIFN-kappa. PMID- 16261203 TI - [Biological activities of recombinant human interferon Epsilon]. AB - OBJECTIVE: To construct a novel recombinant rhIFN-epsilon155ser, and study its biological activities. METHODS: The whole sequence of rhIFN-epsilon was artificially synthesized and some codons were altered according to the preferred codon using of E.coli. The sequence was cloned into plasmid vector pBV220 to express in E.coli DH5alpha. After purification and re-folding of rhIFN epsilon155ser inclusion body, the final product was tested for its biological activities, including anti-viral, anti-proliferative and NK cell enhancing activities. At the same time, by using DNA microarray biochips, the gene expression patterns in the rhIFN-epsilon155ser and rhIFN-alpha2b treated cells were compared and analyzed. RESULTS: The re-built rhIFN-epsilon155ser sequence was expressed in E.coli as a form of inclusion body. After purified and re folded, the rhIFN-epsilon155ser protein reached a purity of above 95%. The rhIFN epsilon155ser protein had a specific anti-viral activity of about 6 x 10(5) IU/mg in WISH/VSV system. Its anti-proliferative activity and NK cell enhancing activities in vitro seemed to be lower than that of rhIFN-alpha2b. Data obtained from microarray biochips indicated that there were 283 pieces increasing 2 folds and 1489 pieces decreasing 2 folds among totally 22,278 pieces of human genes were found in the rhIFN-epsilon155ser treated cells; more changes in gene expression pattern were detected in the rhIFN-alpha treated cells. CONCLUSION: A novel recombinant rhIFN-epsilon155ser was constructed, which belonged to type 1 interferon. The biological activities of rhIFN-epsilon155ser were compared with rhIFN-alpha2b. The changes of gene expression pattern in the interferon treated cells were detected, analyzed and discussed. PMID- 16261204 TI - [Expression, purification and antiviral activities of a new recombinant human interferon-lambda2]. AB - OBJECTIVE: To express recombinant human interferon lambda2 in E.coli and to study its antiviral activities. METHODS: According to preferred codons used in E.coli, the highly-expressed human interferon lambda2 gene was designed, synthesized and cloned into expression vector pBV220 and transfected into E.coli DH5alpha. The expressed product was purified by using CM FF and size exclusion chromatography. Its antiviral activities were tested on different cells. RESULTS: The expressed product was calculated about 15% of the total E.coli protein. The purified protein reached about 90% purity. Its specific antiviral activity was about 1.5 x 10(6) IU/mg on WISH/VSV test system. It was shown that the antiviral activity of the product on primates-origin cells seemed to be much higher than that on other non-primates-origin cells, indicating that interferon lambda2 possessed more stringent species specificity as compared with interferon-alpha2b. New interferon lambda2 showed similar anti-HBV activity as interferon-alpha2b. CONCLUSION: Recombinant human interferon lambda2 could be expressed on E.coli. The purified product showed more stringent species specificity and similar anti-HBV activity as compared with interferon-alpha2b. PMID- 16261205 TI - [Establishment of a new low-density cDNA macroarray and the application in the activity of IFN against HBV]. AB - OBJECTIVE: To investigate the expression profile of genes which are involved in IFN antiviral activity and IFN signal transduction pathway in Hep G2 and HepG2.2.15 cells. METHODS: Genes of interest were selected from the UniGene database (http://www.ncbi.nlm.gov/UniGene/Hs.Home.html). The 5'IMAGE clones with 0.5-0.8 kb length were chosen and ordered from RZPD company. The cDNA inserts were amplified by PCR and then were spotted onto the Hybond-N+ membranes. The membranes were denatured and neutralized for Macroarray analysis. HepG2.2.15 and Hep G2 cells were treated without or with IFN-alpha for 6 h, and the total cellular RNA was isolated using Trizol Reagent. Radio-labelled cDNA was generated from 20 microgram of RNA by reverse transcription using 360 units of reverse transcriptase in the presence of 30 microCi of alpha-32P dCTP. Hybridization was performed between 32P-labelled cDNA and membrane arrays. The membranes were then scanned, and the intensity of autoradiographic spots was quantitated by Cyclone Storage Phosphor System. The images were subsequently analysed by the OptiQuant Imager Analysis Software and converted into digital data. RESULTS: The authors found that just partially IFN-inducible genes were expressed in Hep G2 and HepG2.2.15 cells, and the majority of IFN-inducible genes was lowly responsive or non-responsive to IFN-a treatment. Some interferon-stimulated genes (ISGs) were inhibited or blocked, especially in HepG2.2.15 cells. Interestingly, the authors found that the IFN signal transduction pathway (Jak-STAT) was intact and unimpaired in HepG2.2.15 cells. CONCLUSION: Differential gene expression profiles in response to IFN were found between Hep G2 and HepG2.2.15 cells. PMID- 16261207 TI - [Potential antiviral drug Pueraria crude extract and puerarin protect against ethanol-induced cytotoxicity in embryonic mouse hippocampal cultures]. AB - OBJECTIVE: To examine whether Chinese medical herb Pueraria crude extract (CP) and standard of pure puerarin (SP) possess the same neuroprotective effects during concomitant ethanol (EtOH) treatment. METHODS: Hippocampus cultures were prepared from mice at gestational age of 18 day. Cell viability was measured by MTT assay. RT-PCR was employed to determine mRNA expression of superoxide dismutase (SOD). RESULTS: As measured by MTT assay, supplementation with 15 mg/L CP or 10 mg/L SP afforded neuroprotection against all EtOH concentrations (50, 200 and 350 mmol/L, respectively) in embryonic hippocampal culture system. In addition, both 15 mg/L CP and 10 mg/L SP could decrease expression of SOD at mRNA level. CONCLUSION: This study suggests that CP and SP could decrease oxidative stress induced by ethanol treatment by the decreased expression of SOD at mRNA level, and demonstrates antioxidative neuroprotective effect of CP and SP against developmental ethanol exposure in vitro. PMID- 16261206 TI - [Construction and identification of non-replication recombinant vaccinia virus co expressing human papillomavirus type 16 L1/L2/E6/E7 proteins]. AB - OBJECTIVE: To generate a human papillomavirus (HPV16) prophylactic and therapeutic vaccine candidate for cervical cancer. METHODS: HPV16 major capsid protein L1 gene/minor capsid protein L2 gene and HPV16 early E6/E7 genes were inserted into a vaccinia virus expression vector. A strain of non-recombinant vaccinia virus containing the sequences was obtained through a homologous recombination and identified. RESULTS: DNA hybridization confirmed that the HPV16L1/L2/E6/E7 genes were integrated into vaccinia virus DNA. Western Blot result showed that full-length L1/L2/E6/E7 proteins were co-expressed in CEF cells infected with the recombinant virus. CONCLUSION: NTVJE6E7CKL1L2 could be taken as a candidate of prophylactic and therapeutic vaccine for HPV-associated tumors and their precancerous transformations. PMID- 16261209 TI - [Multiplex RT-PCR for the rapid detection of influenza virus types and subtypes]. AB - OBJECTIVE: To establish a sensitive and specific multiplex RT-PCR(MRT-PCR) for the simultaneous detection of influenza virus types and subtypes. METHODS: Primers were designed from highly conserved region of the hemagglutinin(HA) gene of influenza H1N1H3N2 and B virus and MRT-PCR was performed. Additional two pairs of primers were designed to determine the N1 and N2 subtypes of neuramidinase NA of influenza H1N1 and H3N2 virus. RESULTS: The fragments of HA gene of all types of influenza virus were amplified and there was no cross reaction. The sensitivity of detection of influenza H1N1 and H3N2 virus was 0.10 TCID50/50 microliter by the second PCR and that was 0.01 TCID50/50 microliter for influenza B virus. The NA gene of influenza H1N1 and H3N2 virus was also amplified by this method. CONCLUSION: The sensitivity of detection of influenza virus from clinical patients' throat washing specimens by MRT-PCR was higher than that by MDCK cell culture or egg embryo isolation. This method was highly sensitive and timely for detection of influenza virus types and subtypes. PMID- 16261208 TI - [Screening and cloning of hepatitis C virus non-structural protein 4B interacting protein gene in hepatocytes]. AB - OBJECTIVE: To investigate biological functions of non-structural protein 4B (NS4B) of hepatitis C virus (HCV), yeast-two hybrid technique was performed to seek proteins in hepatocytes interacting with HCV NS4B. METHODS: HCV NS4B bait plasmid was constructed by ligating the NS4B gene with carrier plasmid pGBKT7 and transformed into yeast cells AH109 (type alpha). The transformed yeast cells were amplified and mated with yeast cells Y187 (alpha type) containing liver cDNA library plasmid pACT2 in 2 x YPDA medium. Diploid yeast cells were plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) and synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing x-alpha-gal for selecting two times. After extracting plasmid from blue colonies, plasmid DNA was transformed into competent Escherichia coli and analysed by DNA sequencing and bioinformatics. RESULTS: Five genes in eight positive colonies were obtained. There were one NADH dehydrogenase subunit 3, one cytochrome c oxidase subunit III, one retinol binding protein 4, one reticulon 3-A (RTN3) and one fibrinogen gamma polypeptide (FGG). CONCLUSION: Genes of HCV NS4B interacting proteins in hepatocytes were successfully cloned and the results paved the way for studying the biological functions of NS4B and associated proteins. PMID- 16261210 TI - [Effect of CCR5delta32, CCR5m303, CCR2-64I and SDF1-3'A gene polymorphism on the prognosis of Chinese HIV-1 carriers]. AB - OBJECTIVE: To study how CCR5delta32, CCR5m303, CCR2-64I, SDF1-3'A gene polymorphisms affect the prognosis of Chinese HIV-1 carrier. METHODS: Epidemiologic survey was done to the HIV-1 carriers who were found in Shenzhen area. PCR/RFLP technology was applied to analyze CCR5delta32, CCR5m303, CCR2-64I, SDF1-3'A gene polymorphisms of the HIV-1 carriers. The plasma virus load and CD4+ cell counting was assayed. The incubation period of some carriers was estimated. SPSS11.0 software was used to analyze the data. RESULTS: No persons with CCR5delta32 and CCR5m303 mutation genotype were found from 189 HIV-1 carriers. SDF1-3'A allele frequency was 26.14% and CCR2-64I allele 19.82%. The carriers were divided into high virus load group (virus load < 20,000 copies/ml) and low virus load group (virus load > or =20,000 copies/ml). It was found by one-way ANOVA analysis on the logarithm of virus load that there was no significant difference between CCR2-64I wild genotype and cross bred genotype (P=0.272). One way ANOVA analysis on delitescence of some carriers showed that there was not significant difference between CCR2-64I wild genotype and cross bred genotype (P=0.662). One-way ANOVA analysis on the logarithm of virus load showed that there was significant difference among SDF1-3'A wild genotype, cross bred genotype and pure mutation genotype (P=0.001). CONCLUSION: CCR2-64I gene mutation may not significantly affect virus load of Chinese HIV-1 carriers, nor it affect the incubation period of HIV-1 carriers. SDF1-3'A gene mutation can decrease virus load, but it may not prolong the incubation period of HIV-1 carriers. PMID- 16261211 TI - [Cloning of ACE-2 gene encoding the functional receptor for the SARS coronavirus and its expression in eukaryotic cells]. AB - OBJECTIVE: Angiotensin-converting enzyme 2 (ACE-2) has been identified as a functional receptor of severe acute respiratory syndrome coronavirus (SARS-CoV), so its gene was cloned and eukaryotic expressed for further insight into mechanisms in SARS-CoV entry and pathogenesis, as well as development of a safe and reliable neutralization assay for SARS-CoV. METHODS: Total RNA was extracted from right atrial tissue of a patient with right heart failure resected during a valvular replacement surgery by Trizol one-step method, and the full-length ACE-2 encoding gene was acquired by RT-nested-PCR. The ACE-2 encoding gene was then cloned into pcDNA4/HisMax-TOPO eukaryotic expression vector to construct the recombinant plasmid pcDNA4/ ACE-2, which was then transfected into 293 T cell and ACE-2 eukaryotic transient expression was detected by Western Blot. Syncytia inhibition assay was established to detect SARS-CoV neutralizing antibody, and compared parallelly with SARS pseudovirus neutralization assay. RESULTS: The recombinant plasmid pcDNA4/ ACE-2 could express ACE-2 protein in eukaryotic cells and induce cell-cell fusion between S protein- and ACE2-expressing cells. This cell-cell fusion assay could be used to detect SARS-CoV neutralizing antibody. CONCLUSION: SARS-CoV receptor ACE-2 gene was successfully cloned and eukaryotic expressed, and used to establish syncytia inhibition assay for SARS-CoV neutralizing antibody assay. PMID- 16261212 TI - [Correlation of pathology in chronic hepatitis B to viral markers in serum and hepatic tissue]. AB - OBJECTIVE: To investigate the relation of the viral markers in serum and those expressed by hepatocytes to pathological lesions of hepatic tissue in patients with chronic hepatitis B. METHODS: The relation of viral markers including HBsAg, HBsAb, HBeAg, HBeAb, HBcAb and HBV DNA in serum of 647 patients with chronic hepatitis B and HBsAg, HBcAg expressed by hepatocytes in 418 of these patients to pathological lesions of hepatic tissue was determined. RESULTS: Viral markers in serum and those expressed by hepatocytes in patients with chronic hepatitis B were closely correlated with pathological lesions of hepatic tissue. CONCLUSION: The degree of inflammation and fibrosis in hepatic tissue is milder in serum HBsAg, HBeAb, HBcAb positive and HBV DNA negative patients but more serious in those with negative hepatocytic expression of HBsAg and HBcAg. HBV DNA is not significantly associated with pathological lesions of hepatic tissue. PMID- 16261214 TI - [Screening of the genes of hepatitis B virus e antigen interacting proteins]. AB - OBJECTIVE: To screen and clone the genes in hepatocytes which encode protein that can interact with hepatitis B e antigen(HBeAg) by yeast-two hybridization. METHODS: Recombined HBeAg bait plasmid (pGBKT7-eAg) was transformed into yeast AH l09, followed by mating with yeast Yl87 containing liver cDNA library plasmid in 2 x YPDA medium. Diploid yeast was plated on synthetic dropout nutrient medium (SD/-Trp-Leu-Ade-His) which contains X-a-gal for selecting positive blue clones. Then positive clones were selected and plasmids were prepared and sequenced. Finally, bioinformatics analysis was performed. RESULTS: Totally 245 positive colonies were selected and 101 colonies were sequenced. Through sequences alignment, 6 novel genes and 35 recorded genes were screened. CONCLUSION: Genes of HBeAg interacting proteins have been cloned successfully, which brings some new clues for further studies on the biological functions of HBeAg and the related proteins. PMID- 16261213 TI - [Selection of recombinant fowlpox virus coexpressing HIV-1 gag-gp120 and IL-6]. AB - OBJECTIVE: To construct the recombinant fowlpox virus (rFPV) coexpressing HIV-1 gag-gp120 and hIL-6. METHODS: The recombinant expressing plasmid pUTA-GE-IL6 was successfully constructed by inserting gag-gp120 gene and hIL-6 gene into the downstream of the combined promoter ATI-p7.5 and p7.5 tandem promoter respectively. After transfecting the plasmid into chicken embryonic fibroblast (CEF) cells preinfected with FPV 282E4 strain and selecting the recombinant virus under the pressure of BUdR. The recombinant virus was analyzed by nucleic acid probe hybridization and immunoblotting. In addition, the formation of virus-like particle and the expression of interested proteins in the recombinant virus infected p815 cells were observed, and the immunogenicity of the recombinant virus was also analyzed. RESULTS: There was colorable dot for the positive recombinant virus, immunoblotting analysis showed that the recombinant virus could expressed both gag-gp120 and IL-6. Virus-like particles (VLP) were formed in virus-infected cells, and the interested proteins could be expressed in mammalian cells infected by the recombinant virus. The immunity index from the immunized mice showed that the recombinant virus had good immunogenicity. CONCLUSION: The recombinant fowlpox virus coexpressing gag-gp120 and IL-6 was successfully constructed, which may provide basis for the preparation of live vector genetic engineering vaccine and macromolecule particle vaccine against HIV 1. PMID- 16261215 TI - [Cloning, purification, and antigenic characterization of three recombinant fragments derived from SARS-CoV S1 domain]. AB - OBJECTIVE: The present study aimed to clone and express three fragments of genomic RNA derived from SARS associated coronavirus (SARS-CoV) S1 domain and to study its immunogenicity. METHODS: The S1 domain gene was amplified by PCR with specific primers and was inserted into the prokaryotic expression vector pQE-30. Three fragments (40-751, 746-1344 and 746-2001 bp) derived from S1 domain produced after the recombinant plasmid (pQE-30/S1) was digested by restriction endonucleases. The three fragments were cloned into pQE-30 and expressed in M15 strains of Escherichia coli. The expression products, designated S1a, S1b and S1c respectively, were purified by Ni affinity chromatography. The immunogenicity was analyzed by Western Blot and ELISA using serologically confirmed sera from SARS patients and the sera from healthy donors was used as control at the same assay. RESULTS: Three recombinant plasmids (pQE-30/S1a, pQE-30/S1b, pQE-30/S1c) were constructed.Fusion proteins with relative molecular mass of 26,700, 22,500 and 46,000 dalton were successfully expressed with amounts of 35%, 35% and 30% of total cell protein and purified by Ni affinity chromatography, respectively. Western Blot and ELISA analysis showed that the S1c protein could be specifically recognized by the sera from SARS patients. CONCLUSION: The recombinant S1c protein was a good immunogen and has the potential to be used as a vaccine against SARS-CoV infection. PMID- 16261216 TI - [Cytokines secretion by peripheral blood mononuclear cells from hepatitis C patients after stimulation with synthetic peptides at the highly variable region]. AB - OBJECTIVE: Test the cytokines secretion by peripheral blood mononuclear cells (PBMC) from hepatitis C patients after stimulation with highly variable region (HVR1) synthetic peptides. METHODS: ELISA was used to test the cytokines secretion, flow cytometry to group the proliferated cells, and the antigenicity of synthetic peptide was predicted by the computer modeling. RESULTS: (1)There were PBMC proliferation when stimulated by HVR1 antigen synthetic peptides. (2) There was a rise of IFN-gamma, IL-4 and IL-10. But no rise of IL-2 and TNF-gamma was found. (3) The proliferated cells were mainly CD4+ lymphocytes. CONCLUSION: PMBC from hepatitis C patients tend to secret Th2 cytokines after stimulation with HVR1 designed by the authors. PMID- 16261217 TI - [Sequence-dependent cleavage of HBV DNA by combining with triple helix-forming oligodeoxyribonucleotides modified with manganese porphyrin in vitro]. AB - OBJECTIVE: To observe the ability of triple helix-forming oligonucleotides (TFO) modified with manganese porphyrin to combine with and cleave HBV DNA fractions. METHODS: The ends of TFO were modified with manganese porphyrin and acridine; At 37 degrees C and pH 7.4 condition in vitro, TFO modified with manganese porphyrin and acridine were bound with 32P labeled HBV DNA fragments, the affinity and specificity binding to target sequence were tested by electrophoretic mobility shift and DNase 1 footprinting assays, the ability to cleave HBV DNA fractions was observed with cleavage experiments. RESULTS: TFO modified with manganese porphyrin and acridine could bind to target sequence in a sequence-dependent manner with Kd values of 3.5 x 10(-7) mol/L and a relative affinity of 0.008. In the presence of KHSO5, TFO modified with manganese porphyrin and acridine could cleave target sequence in the region forming triple DNA. CONCLUSION: In the presence of KHSO5, TFO modified with manganese porphyrin and acridine could cleave target HBV-DNA in sequence-dependent manner. PMID- 16261218 TI - [Detection of beef contaminated by bovine central nervous system tissue by enzyme immunoassay]. AB - OBJECTIVE: To study influencing factors of detection of bovine central nervous system (CNS) tissue contaminated beef by enzyme immunoassay (EIA), and the method was applied to the detection of imported beef and domestic beef of China. METHODS: Raw beef homogenates containing different concentrations of raw CNS tissue and the same samples which were heated were detected after different time by RIDASCREEN(r) Risk Material 10/5 and RIDASCREEN(r) Probennahme- zubehor Sampling tools kits. PBS suspension and sample dilution buffer (SDB) suspension of bovine brain tissue with the same concentration of the standard were detected. Beef from USA and domestic market of China were then detected by the kits. RESULTS: The kits could detect both raw and heated CNS tissue in the products with high sensitivity. The absorbance values (AV) increased with the concentrations of CNS in samples. Heating and increasing of time could decrease the absorbance values of the samples which contain CNS tissue. The AV of the PBS suspension of bovine brain tissue was higher than the SDB suspension and the AV of both were higher than the AV of standard of the same concentration. No CNS tissue was detected from all imported beef. No CNS tissue was detected in all samples from domestic market of China except for foxtail. CONCLUSION: The EIA method has high sensitivity for detection of bovine CNS tissue contaminated beef with the glial fibrillary acidic protein (GFAP) as accurate target substance. Heating and increasing of time can lead to decreasing of the AV of samples. Improper slaughter process can lead to contamination of bovine products by bovine CNS tissue. PMID- 16261220 TI - [Histological study of livers from the patients with chronic hepatitis B treated with bicyclol]. AB - OBJECTIVE: To study histological changes of the livers in patients with chronic viral hepatitis B treated with bicyclol tablets. METHODS: Thirty one patients with chronic viral hepatitis B were divided into two groups and were treated with bicyclol orally at doses of 150 mg daily or 75 mg daily for 36 weeds. The histological changes of the livers were observed before and after the treatment. RESULTS: Compared with pre-treatment findings, there were significant differences in histological activity index in each group (P < 0.01, P < 0.05), there were also significant differences between the two groups (P < 0.05). Decreased inflammatory reaction was also seen (P < 0.05). CONCLUSION: Daily use of 150 mg and 75 mg bicyclol tablets are effective in improving liver histological changes in chronic hepatitis B patients. Bicyclol 150 mg daily was better. PMID- 16261219 TI - [Experimental study on the inhibitory effect of Carboxymethyl Pachymaram on hepatitis B virus expression from transfected cells]. AB - OBJECTIVE: To observe the anti-HBV activity of Carboxymethyl Pachymaram (CMP) on the culturing of 2.2.15 cell line. METHODS: Concentrations of 20.0 g/L, 12.0 g/L, 6.0 g/L, 3.0 g/L, 1.5 g/L of CMP were used to evaluate its toxicity to the cell line and the inhibition rates of the secretion of HBsAg and HBeAg in the cultured 2.2.15 cell line. RESULTS: Experiments showed that the mean half toxicity concentration of CMP for 2.2.15 cell line was 13.6 g/L and concentration for 50% inhibition of the secretion of HBsAg and HBeAg were 4.45 g/L, 5.61 g/L and TI were 3.06 and 2.42. CMP showed stronger effect on anti-HBV than aciclovir. CONCLUSION: CMP has good inhibitory effect on the secretion of HBsAg and HBeAg on cultured cell line 2.2.15. PMID- 16261221 TI - Contemporary superconducting materials. AB - Superconductors currently under active study are briefly described from a chemical perspective. The author's current views on those superconductors, the role of theory in searching for new superconductors, and possibilities for new superconducting materials that might be found in the future are presented. PMID- 16261222 TI - Synthesis, processing and properties of conjugated polymer networks. AB - Despite the diverse research activities focused on the chemistry, materials science and physics of conjugated polymers, the feature of conjugated cross links, which can provide electronic communication between chains, has received little attention. This situation may be a direct consequence of the challenge to introduce such links while retaining adequate processability. Focusing on recent studies of materials for which charge transport or electrical conductivity data are available, this feature article attempts to present an overview of the synthesis, processing and electronic properties of conjugated polymer networks. For the purpose of this discussion, two distinctly separate architectures featuring covalent cross-links on the one hand and non-covalent organometallic bridges on the other-are treated in separate sections. The available data indicate that cross-linking can have significant benefits for intermolecular charge transfer if the polymers are carefully designed. PMID- 16261223 TI - Template synthesis of tungsten complexes with saturated N-heterocyclic carbene ligands. AB - Tungsten complex with a coordinated 2-azidoethyl isocyanide ligand reacts with PMe3 at the azido function to give a complex with a coordinated iminophosphorane which upon hydrolysis of the P=N bond yields a complex with an NH,NH-stabilized N heterocyclic carbene ligand, 7; alkylation of the carbene ring nitrogen atoms gives a complex with an N,N'-dialkylated imidazolidin-2-ylidene ligand, 8 . PMID- 16261224 TI - Vibrational circular dichroism of N-acetyl-l-cysteine protected gold nanoparticles. AB - Vibrational circular dichroism is used to determine the conformation of a thiol adsorbed on gold nanoparticles. PMID- 16261225 TI - Non-stoichiometry induced by differential oxygen/lone pair occupation in chiral bicyclic 1,1'-binaphthoxy cyclodiphosphazanes. AB - Non-stoichiometry and isostructurality in a set of chiral phosphorus compounds as a result of lone pair/oxygen exchange, substantiated by the combined use of 31P NMR spectroscopy and X-ray crystallography, is described. PMID- 16261226 TI - A novel route for in-situ H2O2 generation from selective reduction of O2 by hydrazine using heterogeneous Pd catalyst in an aqueous medium. AB - Hydrogen peroxide in high yields can be generated with high efficiency at mild conditions (25 degrees C and atmospheric pressure) with the formation of only environment-friendly by-products (N2 and H2O) by a reduction of O2 by hydrazine from its hydrate/salt with its complete conversion in a short reaction period (or=4 adjacent, with 2-3 adjacent and with 1 single cold segments were named as Group1(GR1), Group2(GR2) and Group3(GR3), respectively. Secondly, the patients were re-grouped according to RVG-EF values. (Group A: patients with EF<50%; Group B: patients with EF>or=50%). In each group, the GPS-EFs were compared with RVG performed within one week and also the variations of GPS-RVG EF differences among the groups were statistically analyzed. RESULTS: In overall (r=0.86) and in each subgroup, EFs obtained by GPS were well correlated with RVG. However, in overall (difference mean EF% [dEF%]=4.6+/-6.7, p<0.001) as well as in subgroup evaluation, GPS significantly (p<0.005) underestimated EF. There was no statistically significant difference in GPS-RVG EF variations between GR1, GR2 and GR3 (p>0.05). The RVG-mean differences and RVG-correlation coefficients calculated for GR1,GR2 and GR3 were dEF%=3.1+/ 4.6, r=0.85; dEF%=3.7+/-6.03, r=0.80 and dEF%=6.2+/-8.03, r=0.79, respectively. Mean dEF% was statistically higher in group-B than group-A (mean difference of dEF%=4,2, p<0.05). In group-A, GPS-EF values were better agreed with RVG (dEF%=3.34, r=0.75) than in group-B (dEF%=7.52, r=0.53). CONCLUSION: The stability of the calculation algorithm of QGS in EF calculation of patients with large depleted infarct areas could be confirmed. The agreement of GPS determined EF is higher in patients having myocardial integrity loss and left ventricular dysfunction. PMID- 16261280 TI - The long-term predictive value of bone mineral density measurements for fracture risk is independent of the site of measurement and the age at diagnosis: results from the Prospective Epidemiological Risk Factors study. AB - Although the utility of bone mass measurements has been the subjects of extensive investigations, the number of studies comparing the predictive value of bone mass measurement at different skeletal sites in the same cohort with a large number of clinically verified endpoints is limited. Furthermore, scant information is available on how age at the time of diagnosis influence the risk of future fractures posed by low bone mineral density (BMD). We have followed 5,564 Danish postmenopausal women for a mean period of 7.3 years. Bone mineral content (BMC) at the forearm and BMD at the spine and hip were assessed at baseline. Vertebral fractures were assessed on digitalized images of lateral X-rays of the thoracic and lumbar spine, whereas non-vertebral fractures were self-reported. At follow up, 17.6% of the women revealed an incidental vertebral fracture and 14.2% reported a new non-vertebral fracture. The absolute risk per 1,000 person-years of osteoporotic fracture increased significantly with decreasing bone mass at all three skeletal sites (P<0.001). Osteoporotic BMD (T-score < or =-2.5) had similar predictive values of fractures regardless of the skeletal site of measurement. Furthermore, the absolute risk of osteoporotic fractures increased significantly with increasing age at the same level of bone mass. Interestingly, the relative risk (RR) of vertebral fracture accompanying 1 SD decrease in spine BMD was similar across different age groups: <55 years (RR:2.1, 95% CI 1.3-3.3), 55-64 years (RR:2.3, 95%CI 1.7-3.2), 65-74 years (RR:2.0; 95%CI 1.5-2.6). Furthermore, women with any prior osteoporotic fracture had a 2.4-fold (95% CI 2.01-2.75, P<0.001) increased risk of a new vertebral fracture. Both age and prior fracture are strong predictors of future fractures. The long-term predictive value of bone mass measurement is independent of the site of measurement and the age at diagnosis. PMID- 16261281 TI - CNS involvement occurs more frequently in patients with Behcet's disease under cyclosporin A (CSA) than under other medications--results of a retrospective analysis of 117 cases. AB - The aim of this study was to evaluate the incidence of neurological manifestations of Behcet's disease (BD) in patients on cyclosporin A (CSA) compared with those on other medications. The records of 117 patients with BD who visited our hospital between 1990 and 2003 were reviewed with respect to symptoms and medication. All episodes of constant therapy prior to central nervous system (CNS) involvement were counted, and then the associations were analysed by the exact Fisher-Freeman-Halton test and adjusted for multiple tests by the Bonferroni-Holm method. We observed ten new cases of CNS manifestations in our patients with BD being regularly seen and treated in our tertiary care centre. The overall prevalence of neuro-BD in our patient group was 8.5%. In a retrospective analysis, the incidence of new-onset neurological disease (neuro BD) in all patients with BD who regularly visited our hospital was significantly higher in patients on CSA than in those on other medications (6 of 21 vs 0 of 175 episodes, P<0.0001). This contrasts the obvious efficacy of CSA on extracerebral manifestations of BD, such as severe ocular disease, mucocutaneous lesions or arthritis. CSA exerts differential efficacy on various manifestations of BD. It is very effective for severe ocular and other moderate to severe manifestations of BD, but its efficacy for the prevention of neuro-BD seems to be inferior to that of other medications used in BD, such as azathioprine or interferon-alpha. The reasons for this are unclear, but the potential toxic effects of CSA on the CNS may be a predisposing factor for CNS vasculitis in BD. PMID- 16261282 TI - Complete heart block in a patient with systemic sclerosis. AB - We describe a patient with diffuse systemic sclerosis and presyncopal episodes where the electrocardiogram revealed complete atrioventricular heart block associated with left posterior and right heart bundle block. The patient underwent implantation of a permanent pacemaker. PMID- 16261283 TI - Tryptophan degradation increases with stage in patients with rheumatoid arthritis. AB - Immune system activation is known to be involved in the progression of rheumatoid arthritis (RA). The proinflammatory cytokine interferon-gamma in various cells, including monocytes, induces the enzyme indoleamine (2,3)-dioxygenase (IDO), which converts tryptophan to kynurenine. In sera of 22 patients (17 women and 5 men) with RA stages 1 to 4 according to Steinbrocker, the concentrations of tryptophan and kynurenine were measured by high-pressure liquid chromatography. To estimate IDO activity, the kynurenine to tryptophan ratio (kyn/trp) was calculated. In parallel, concentrations of the macrophage activation marker neopterin were determined by enzyme-linked immunosorbent assay. Tryptophan concentrations were lower in patients with RA, and the decrease in serum tryptophan correlated with increase in stage (p<0.05). Kyn/trp correlated well with neopterin concentrations, which were elevated in most patients. Whereas higher C-reactive protein concentrations and erythrocyte sedimentation rates were observed in patients with greater disease activity, tryptophan and neopterin concentrations did not differ between patients with different subjective disease activity graded by the physician. Deficiency of the essential amino acid tryptophan in patients with RA most likely results from immune activation involved in the pathogenesis of the disease. It could also be relevant for the mood of patients, as tryptophan is the precursor of serotonin. PMID- 16261284 TI - Renal amyloidosis: an uncommon complication of juvenile idiopathic arthritis. AB - A 9-year-old girl presented with systemic-onset juvenile idiopathic arthritis, diagnosed at 3.5 of age and which was difficult to control despite several therapeutic trials. Five years after diagnosis of juvenile idiopathic arthritis, nephrotic proteinuria was noticed. Renal biopsy confirmed the diagnosis of amyloidosis, and chlorambucil was initiated, with general improvement of the disease and reduction of proteinuria. PMID- 16261285 TI - Different clinical features in patients with limited and diffuse cutaneous systemic sclerosis. AB - This study aims to analyze differences among established disease damage indicators in patients with limited cutaneous systemic sclerosis (lcSSc) and diffuse cutaneous systemic sclerosis (dcSSc). Fifty patients with lcSSc and 55 patients with dcSSc were included in this study. Difference in mean disease duration between the two subgroups of patients was not statistically significant (z=-0.88, p=0.38). Patients with lcSSc and dcSSc were compared, and differences in vascular, esophageal, lung, heart, renal, and musculoskeletal involvement were statistically assessed using chi (2), Mann-Whitney, and Kruskal-Wallis tests. Using the technique of nailfold capillaroscopy, we found normal capillaries or nonspecific capillary change in 10.0% of the patients with lcSSc and only in 3.6% of the patients with dcSSc. Dilated capillaries without loss of capillaries were found in 42% of the patients with lcSSc and in 10.9% of the patients with dcSSc (p=0.05). However, severe capillary damage (loss of capillaries) was noticed more frequently in patients with dcSSc (dcSSc/lcSSc: 85.5%/48.0%, p=0.002). Pitting scars or digital ulcers were found in 46.0% of the patients with lcSSc and in 67.3% of the patients with dcSSc (p=0.04). We did not notice a significant difference in frequency of fingertip osteolysis and telangiectasia. Esophageal hypomotility was found in 64% of the patients with lcSSc and in 85.5% of the patients with dcSSc (p<0.01). We found interstitial lung fibrosis more frequently in patients with dcSSc (lcSSc/dcSSc: 16.0%/72.7%, p<0.001). Reduced forced vital capacity (FVC) was found in 6.0% of the of patients with lcSSc and in 41.8% of the patients with dcSSc (p<0.001). A decreased value of the transfer factor for carbon monoxide (DLCO) was also observed more frequently in patients with dcSSc. Heart involvement was found in 29.1% of the patients with dcSSc and less frequently (p<0.001) in patients with lcSSc (8%). Similarly, we found renal involvement more frequently in patients with dcSSc (lcSSc/dcSSc: 2.0%/16.3%). Tendon friction rubs were noticed in 23.6% of the patients with dcSSc and only in 6% of the patients with lcSSc (p<0.01). Joint contractures were observed in 70.9% of the patients with dcSSc and in 26.0% of the patients with lcSSc (p<0.001). Muscle weakness was noticed more frequently in patients with dcSSc (lcSSc/dcSSc: 22.0%/40.0%, p<0.05). Arthralgia was found more frequently in patients with dcSSc, but arthritis became apparent, without significant difference in frequency, in 16% of the patients with lcSSc and in 16.4% of the patients with dcSSc. Loss of capillaries (detected by nailfold capillaroscopy), digital ulcers, interstitial lung fibrosis, decreased FVC and DLCO, esophageal hypomotility, musculoskeletal impairment, and heart and renal involvement are more common in patients with dcSSc. Fingertip osteolysis, telangiectasia, and arthritis are equally frequent in both forms of the disease. PMID- 16261286 TI - Ultrasound detection of entheseal insertions in the foot of patients with spondyloarthropathy. AB - The aim of the study was to evaluate entheseal involvement in the foot of patients with spondyloarthropathy (SpA) by ultrasonographic (US) examination and compare the results with clinical and radiological findings. Forty-four patients (27 men, 17 women) with a diagnosis of SpA were recruited. Patient evaluation included physical examination (swelling and pain), Bath Ankylosing Spondylitis Radiological Index, Bath Ankylosing Spondylitis Disease Activity, Bath Ankylosing Spondylitis Metrological Index, and laboratory parameters (erythrocyte sedimentation rate, C-reactive protein). Foot x-rays of the patients were assessed and scored according to SpA Tarsal Radiographic Index (SpA-TRI). A high resolution US was used to examine the tendon and ligament insertion sites in the foot, and the findings were scored according to Glasgow Ultrasound Enthesitis Scoring System (GUESS). The mean age and disease duration of the patients were 39.9+/-12.5 and 9+/-8.2 years, respectively. US revealed pathological findings in 25 of 44 (56.8%) patients, most of whom exhibited no clinical signs of foot involvement. Pain and swelling at entheseal insertions were detected in only 16 (37%) patients. The mean GUESS score and SpA-TRI score were 2.2+/-2.5 and 3.3+/ 3.7, respectively. There was a correlation between the scores of GUESS and SpA TRI, particularly at the Achilles and plantar fascia insertion sites. The mean score of SpA-TRI was higher in patients with enthesopathy, detected by US, than in patients without enthesopathy (4.6+/-4.4 vs 1.8+/-2.1). There was no significant correlation between the mean GUESS score and clinical and laboratory variables. In conclusion, involvement of tendon and entheses is not rare, and US is a valuable diagnostic method in detecting subclinical enthesopathic changes. The GUESS and SpA-TRI are suggested in quantifying US and radiological changes in the foot of patients with SpA. PMID- 16261287 TI - A randomized, double-blind, placebo-controlled study of the safety and efficacy of intravenous MCC-135 as an adjunct to primary percutaneous coronary intervention in patients with acute myocardial infarction: rationale and design of the evaluation of MCC-135 for left ventricular salvage in acute MI (EVOLVE) study. AB - As a consequence of acute ischemia and reperfusion in patients with acute ST elevation myocardial infarction, calcium overload inside myocytes not only affects myocardial contraction, relaxation, and myocyte recovery following reperfusion, but also may be related to myocyte necrosis and fatal arrhythmia. MCC-135 is the first in a new class of agents that reduce intracellular calcium overload. Pre-clinical and early clinical studies yielded promising results for patients with ST elevation myocardial infarction. The Evaluation of MCC-135 for Left Ventricular Salvage in Acute MI (EVOLVE) study is a Phase 2a, multicenter, randomized, double-blind, placebo-controlled clinical trial of 2 new doses of MCC 135 (4.5 mg/kg/48 hours and 9.0 mg/kg/48 hours) as adjunct therapy for preservation of left ventricular function and reduction of infarct size in patients undergoing primary percutaneous coronary intervention (PCI) for electrocardiographically moderate-large ST elevation myocardial infarction. The primary endpoint will be left ventricular ejection fraction on Day 5 post myocardial infarction as determined by single photon emission computed tomography (SPECT). Secondary endpoints will include SPECT and echocardiographic assessments, serum cardiac markers, clinical outcomes, and safety measures at specific time points through Day 30 post myocardial infarction. Follow-up clinical and safety assessments will be continued until Day 180. The rationale, design, and methods of the EVOLVE study are described in this paper, along with 2 sub-studies, involving a comparison of pre- and post-PCI measurements with either SPECT or echocardiography, to examine myocardial salvage and the time course of changes in myocardial infarction size and left ventricular function. MINIABSTRACT: The Evaluation of MCC-135 for Left Ventricular Salvage in Acute MI (EVOLVE) study is a Phase 2, multicenter, randomized, double-blind, placebo controlled clinical trial of two doses of MCC-135, first in a new class of agents that reduce intracellular calcium overload, as adjunct therapy for preservation of left ventricular function and reduction of infarct size in patients with moderate-large STEMI undergoing primary PCI. The rationale, design, and methods of the EVOLVE study, along with two sub-studies, are described in this paper. PMID- 16261288 TI - Combined thrombolysis with abciximab favourably influences platelet-leukocyte interactions and platelet activation in acute myocardial infarction. AB - BACKGROUND: In patients with acute myocardial infarction (AMI), activated platelets and altered haemostatic/fibrinolytic systems with and without thrombolytic therapy are known. Platelets thereby interact with neutrophils, stimulated endothelial cells and with monocytes leading to adverse effects on further myocardial damage. Thrombolysis in these patients is still hampered by procoagulant effects favoring early reocclusion. The additional treatment with a GPIIb/IIIa antagonist aimed to minimize early reocclusion thus improving the present therapeutic regimen. METHODS: In 38 patients with AMI, we investigated the effects of a thrombolytic regimen with half reteplase (r-PA) dose plus abciximab vs. full dose r-PA on membrane-bound adhesion molecules (CD41, CD42b, CD40, CD40L) expressed on platelets, neutrophils and monocytes as well as on soluble platelet-selectin as interaction and activation markers of these cells. RESULTS: The combination group had significantly (p < 0.05) lower sP-selectin levels over 48 h vs. the group treated with full dose r-PA. After 3 h, the percentage of CD41 and CD42b positive monocytes and granulocytes as well as the percentage of CD40 positive granulocytes and the percentage of CD40L positive monocytes markedly (p < 0.01, p < 0.05) decreased in the combination group vs. data at admission compared with the r-PA group indicating less leukocyte-patelet adhesion. CONCLUSIONS: The thrombolytic regimen with half dose r-PA and abciximab had a benefical influence on platelet activation and induced a more marked decrease of platelet-monocyte, and in part, platelet-granulocyte aggregates compared with the r-PA regimen. This could contribute to a probably lesser monocyte activation state with favourable effects on monocyte-endothelial adhesion and a consecutively possible influence of myocardial damage, a reduction of the additionally acute local inflammatory processes and a reduction of adherence of platelet-granulocyte aggregates to subendothelium. PMID- 16261289 TI - Varied prevalence of factor V G1691A (Leiden) and prothrombin G20210A single nucleotide polymorphisms among Arabs. AB - BACKGROUND: Factor V G1691A (FV-Leiden) and prothrombin (PRT) G20210A single nucleotide polymorphisms (SNPs) are major inherited risk factors of venous thromboembolism. In view of the heterogeneity in their world distribution and lack of sufficient information about their distribution among Arabs, we addressed the prevalence of both SNPs in 4 distinct Arab populations (Lebanon, Tunisia, Bahrain, and Saudi Arabia). METHODS: Study subjects comprised 698 Lebanese, 313 Tunisian, 194 Bahraini, and 149 Saudi Arabian healthy subjects; genotyping was done by PCR-RFLP using Mnl I and Hind III for FV-Leiden and PRT G20210A, respectively. RESULTS: The prevalence of the mutant A alleles of FV-Leiden and PRT G20210A were significantly higher among Lebanese (0.0788 and 0.0136) and Tunisians (0.0351 and 0.0128), as compared to Bahraini (0.0155 and 0.0052) and Saudi (0.0101 and 0.000) subjects. Higher frequency of the FV-Leiden G/A and A/A genotypes were seen in Lebanon (13.8 and 1.0%), followed by Tunisia (5.8 and 0.6%), Bahrain (3.1 and 0.0%) and Saudi Arabia ((2.0 and 0.0%). All PRT G20210A positive cases were in the heterozygote (G/A) state, and these comprised 3.6% for Lebanon, 2.6% for Tunisia, 1.0% for Bahrain. The carrier rate of FV-Leiden was significantly higher among Lebanese compared to the other populations (p < 0.001), while the difference in the prevalence of FV-Leiden between the other populations was not statistically different. With the exception of Lebanese-Saudi (p = 0.038), the prevalence of PRT G20210A was similar among the study communities. Furthermore, the overall average genetic differentiation between populations (estimated with the F(ST)) was 0.0022 for FV-Leiden and 0.005 for PRT G20210A. CONCLUSIONS: These results further confirm the heterogeneity in FV Leiden and PRT G20210A distribution among Arabs, and recommend potential institution of prophylactic measures for carriers of either or both SNPs. PMID- 16261290 TI - Thrombolysis with streptokinase during cardiopulmonary resuscitation: a single center experience and review of the literature. AB - OBJECTIVE: To report our experience with use of thrombolysis with streptokinase during cardiopulmonary resuscitation of patients with cardiac arrest due to myocardial infarction. DESIGN: A case series. METHODS: Thrombolytic therapy (streptokinase) was administered during cardiopulmonary resuscitation of 4 patients with suspected myocardial infarction as the cause of cardiac arrest. RESULTS: 3 of the 4 patients survived and were discharged from the hospital without any major complications or neurological sequela. CONCLUSION: Thrombolysis with streptokinase during cardiopulmonary resuscitation of patients with suspected acute myocardial infarction is associated with reduced mortality and favorable neurological outcome. PMID- 16261291 TI - Mean platelet volume in the patients with cardiac syndrome X. AB - BACKGROUND: Angina with normal coronary arteries, cardiac syndrome X, is a diagnosis of exclusion. The exact mechanism of this clinical syndrome remains unclear. Although the prognosis is as good as equal to that of normal population, symptoms related with the syndrome impair largely quality of life. Mean platelet volume showing the platelet size is an indicator of platelet function. Larger platelets are more active than smaller ones. METHODS AND RESULTS: We designed a study, evaluated mean platelet volume of the patients with cardiac syndrome X (group A) and stable angina (group B) and investigated the relation between groups. Eighty patients with cardiac syndrome X with a mean age of 51.08 +/- 9.79 years and 67 patients with stable angina with a mean age of 55.16 +/- 11.96 years were studied. At the end of the study, mean platelet volume of group A was significantly higher than that of group B, 10.55 +/- 1.08 fl vs. 9.39 +/- 0.58 fl, respectively (P < 0.001). CONCLUSION: Raised platelet size has been shown to be associated with adverse cardiac events. Mean platelet volume has increased in acute coronary syndromes and also in cardiac syndrome X in our study. Life style modification may optimize platelet size and improve symptoms in these patients. PMID- 16261292 TI - Venous thromboembolism in cancer patients referred to the American University of Beirut-Medical Center secondary to deep vein thrombosis; occurrence and risk factors. AB - BACKGROUND/OBJECTIVES: Cancer is a very important risk factor for the development of thromboembolic disorders. We conducted a retrospective study, in order to examine the rate of Deep vein thrombosis occurrence in cancer patients, as well as the effects of risk factors for DVT in cancer patients affected with DVT. METHODS: 148 patients were enrolled in this study. Several risk factors for DVT were retrospectively examined with the main emphasis being age, gender, obesity, previous history of past thromboembolic phenomena and metastatic disease. RESULTS: 55.5% of the study group was older than 60 years of age. Gender did not appear to be a significant risk factor. As for obesity, only 23% of our patients were defined as obese. An examination of previous history of thromboembolic conditions revealed that 16.8% had previous history of DVT, 1.3% developed a PE prior to presentation and 3.4% were positive for both DVT and PE. As for metastasis, 33.7% of our study population exhibited metastatic cancer upon their presentation. One interesting finding was that only 13 patients out of the 148 had been on DVT-prophylaxis prior to their admission. CONCLUSION: Cancer is an important risk factor for the development of thromboembolic phenomena, yet in most parts of the world, prophylaxis is discerned mostly through clinical judgement or local guidelines. With this in mind, and the additive effects of other risk factors, stricter medical practices and well-defined regimens should be set forth to protect cancer patients from these conditions. PMID- 16261294 TI - Biomarkers in atrial fibrillation: further observations on biologic plausibility, cause and effect. PMID- 16261295 TI - Radiation exposure and coronary atherothrombosis. PMID- 16261293 TI - A descriptive evaluation of routine complete blood count monitoring in patients receiving anticoagulation therapy. AB - CONTEXT: The most prevalent side effect associated with warfarin therapy is bleeding. Routine monitoring of laboratory tests, such as complete blood counts (CBC), may provide theoretical benefit in this patient population as a means of detecting occult bleeding through hemoglobin values that decrease compared to baseline. OBJECTIVE: To describe the clinical outcomes associated with routine CBC monitoring in a large, diverse sample of anticoagulated patients. DESIGN: Forty-seven month retrospective, observational study. SETTING: Large non-profit, group-model health maintenance organization with a centralized clinical pharmacy anticoagulation service that routinely orders baseline CBCs on all patients enrolled in the service with follow up CBCs repeated at 3 months and yearly thereafter. PATIENTS: Patients continuously enrolled in the anticoagulation service for warfarin therapy monitoring for at least one year between January 1, 2000 and December 5, 2003. MAIN OUTCOME MEASURES: Medical records were reviewed to determine clinical outcomes associated with hemoglobin decreases of > or =2.0 gm/dL. Clinical outcomes included both overt and occult bleeding events and a determination of major or minor bleeding in these events. RESULTS: Of the 4033 patients included in the analysis, 578 (14.3%) experienced at least one decrease in hemoglobin > or =2.0 gm/dL. Occult bleeding was confirmed in 121 patients (3.0% of all patients monitored) with a decrease of hemoglobin > or =2.0 gm/dL but only 13 of these patients (0.3% of all patients monitored) experienced major bleeding. The annual detection rate of occult bleeding through routine CBC monitoring was 0.8%. CONCLUSIONS: Although routine CBC monitoring provides some utility in detecting occult bleeding, the yield of clinically important decreases in hemoglobin detected was low. For most anticoagulated patients, routine CBC monitoring appears to be clinically unnecessary. PMID- 16261296 TI - Rearrangements in icosahedral boranes and carboranes revisited. AB - The structure, stability, and intermolecular rearrangements between ortho-, meta , and para-C2B10H12 and were investigated using the hybrid density functional B3LYP/6-31G(d) for vibrational frequencies, as well as B3LYP/6-311+G(2d,p) for single-point electronic energies. The general trends in free energies of rearrangement between ortho-C2B10H12 to meta-C2B10H12 and meta-C2B10H12 to para C2B10H12 presented here are consistent with experimental reaction temperatures. In addition, the majority of the rearrangements can be viewed in terms of concerted diamond-square-diamond steps and triangular face rotations. PMID- 16261297 TI - Periodic trends and easy estimation of relative stabilities in 11-vertex nido-p block-heteroboranes and -borates. AB - Density functional theory computations were carried out for 11-vertex nido-p block-hetero(carba)boranes and -borates containing silicon, germanium, tin, arsenic, antimony, sulfur, selenium and tellurium heteroatoms. A set of quantitative values called "estimated energy penalties" was derived by comparing the energies of two reference structures that differ with respect to one structural feature only. These energy penalties behave additively, i.e., they allow us to reproduce the DFT-computed relative stabilities of 11-vertex nido heteroboranes in general with good accuracy and to predict the thermodynamic stabilities of unknown structures easily. Energy penalties for neighboring heteroatoms (HetHet and HetHet') decrease down the group and increase along the period (indirectly proportional to covalent radii). Energy penalties for a five- rather than four-coordinate heteroatom, [Het(5k)(1) and Het(5k)(2)], generally, increase down group 14 but decrease down group 16, while there are mixed trends for group 15 heteroatoms. The sum of HetHet' energy penalties results in different but easily predictable open-face heteroatom positions in the thermodynamically most stable mixed heterocarbaboranes and -borates with more than two heteroatoms. PMID- 16261298 TI - Theoretical study of hydrogenation of the doubly aromatic B7- cluster. AB - We have studied the influence of hydrogenation on the relative stability of the low-lying isomers of the anionic B7- cluster, computationally. It is known that the pure-boron B7- cluster has a doubly (sigma- and pi-) aromatic C6v(3A1) quasi planar wheel-type triplet global minimum (structure 1), a low-lying sigma aromatic and pi-antiaromatic quasi-planar singlet C2v (1A1) isomer 2 (0.7 kcal mol(-1) above the global minimum), and a planar doubly (sigma- and pi-) antiaromatic C2v (1A1) isomer 3 (7.8 kcal mol(-1) above the global minimum). However, upon hydrogenation, an inversion in the stability of the species occurs. The planar B7H 2- (C2v, 1A1) isomer 4, originated from the addition of two hydrogen atoms to the doubly antiaromatic B7- isomer 3, becomes the global minimum structure. The second most stable B7H2- isomer 5, originated from the quasi-planar triplet wheel isomer 1 of B7- , was found to be 27 kcal mol(-1) higher in energy. The inversion in stability occurs due to the loss of the doubly aromatic character in the wheel-type global minimum isomer (C6v, 3A1) of B7- upon H2-addition. In contrast, the planar isomer of B7- (C2v, 1A1) gains aromatic character upon addition of two hydrogen atoms, which makes it more stable. PMID- 16261299 TI - [Assessment of health-related quality of life with the German SF-8. A comparison of telephone and postal survey modes]. AB - Within a comprehensive comparison of telephone and postal survey methods the SF-8 was applied to assess adult's health-related quality of life. The 1690 subjects were randomly assigned to a telephone survey and a postal survey. Comparisons across the different modes of administration addressed the response rates, central tendency, deviation, ceiling and floor effects observed in the SF-8 scores as well as the inter-item correlation. The importance of age and gender as moderating factors was investigated. Results indicate no or small statistically significant differences in the responses to the SF-8 depending on the actual mode of administration and the health aspect questioned. It was concluded that further investigations should focus on the exact nature of these deviations and try to generate correction factors. PMID- 16261300 TI - [Work disability from mental disorders]. AB - Sick leave certifications allow patients to stay away from work without negative consequences. German regulations state that patients can be granted sick leave when duties can no longer be fulfilled or may endanger health status and when there is a causal relationship between illness and disability. Relevant is not the presence of illness but of a disability which interferes with work requirements. In Germany, an average of 4% of workers are on sick leave at any given time, corresponding to about 2 weeks per year and person. The level of sick leave depends not only on the patient's health status but also on social factors. Mental disorders are a major reason for sick leave. Sick leave can have negative effects by fostering chronicity. In 75% of cases, physicians have been found to grant sick leave incorrectly. It must be kept in mind that not illness per se but only disorders of participation justify sick leave. PMID- 16261301 TI - Successful treatment of Candida glabrata peritonitis with fluconazole plus flucytosine in a premature infant following in vitro fertilization. PMID- 16261302 TI - Fatal varicella zoster virus infection as first manifestation of idiopathic CD4+ T-cell lymphocytopenia. PMID- 16261304 TI - Sensitivity and specificity of a new commercial enzyme-linked immunoassay kit for detecting Entamoeba histolytica IgG antibodies in serum samples. AB - The study presented here was conducted to evaluate the performance of the newly available RIDASCREEN Set (R-Biopharm AG, Darmstadt, Germany) for the detection of immunoglobulin G antibodies against Entamoeba histolytica. The sensitivity and specificity of this new enzyme-linked immunosorbent assay were evaluated using a panel of sera from 239 individuals. The assay was positive for 43 of 44 patients with invasive amebiasis, including all 18 patients with amebic liver abscess, while it was negative for 190 of 195 adult controls who were either healthy individuals or patients with other parasitic diseases. The kit was found to be highly specific (97.4%) and sensitive (97.7%) for detecting antibodies against E. histolytica in humans. Although antibody titers in patients with amebic liver abscess tend to be higher on average than in patients with invasive amebiasis, it is not possible to distinguish the two forms solely based on the results of this commercial test. PMID- 16261303 TI - Antimicrobial resistance of Streptococcus pneumoniae isolates in Athens, Greece. AB - In order to determine the current antibiotic susceptibility of Streptococcus pneumoniae strains in Greece, the present study was performed on 282 clinical isolates collected from patients at the Sotiria Chest Hospital of Athens, Greece, during the years 1997-2003. Susceptibility testing revealed that 52 (18.4%) isolates were not susceptible to penicillin, with 13.1% demonstrating intermediate and 5.3% high-level resistance. One of the penicillin-non susceptible isolates was also resistant to cefotaxime. Comparison with results of a previous study conducted at the same hospital during the period 1992-1993 showed that penicillin resistance had increased by 4.5%. The results of this study indicate the antimicrobial susceptibility of Streptococcus pneumoniae continues to change in Greece and continuous surveillance remains important for guiding empirical antibiotic therapy. PMID- 16261305 TI - Evaluation of the new Vitek 2 GN card for the identification of gram-negative bacilli frequently encountered in clinical laboratories. AB - The new Vitek 2 GN card (bioMerieux, Marcy-l'Etoile, France) was developed for better identification of fermenting and nonfermenting bacilli. This new card allows the identification of 159 taxa. A total of 426 isolates (331 fermenting and 95 nonfermenting gram-negative bacilli) belonging to 70 taxa covered by the database were evaluated. All isolates were identified in parallel with the ID 32 GN, the API 20E, and the API 20NE methods. The system correctly identified 97.4% (n=415) of the strains. Only 2.1% (n=9) needed additional testing. One strain (0.25%) was misidentified (Klebsiella pneumoniae subsp. pneumoniae), and another one (0.25%) was not identified (Morganella morganii subsp. morganii). The new GN card gives more accurate identifications overall for gram-negative bacilli when compared to the systems described in other similar studies. PMID- 16261306 TI - International, open-label, noncomparative, clinical trial of micafungin alone and in combination for treatment of newly diagnosed and refractory candidemia. AB - Candida spp. are the fourth leading cause of bloodstream infections, and non albicans species are increasing in importance. Micafungin is a new echinocandin antifungal agent with excellent in vitro activity against Candida spp. Pediatric, neonatal, and adult patients with new or refractory candidemia were enrolled into this open-label, noncomparative, international study. The initial dose of micafungin was 50 mg/d (1 mg/kg for patients <40 kg) for infections due to C. albicans and 100 mg/d (2 mg/kg for patients <40 kg) for infections due to other species. Dose escalation was allowed. Maximum length of therapy was 42 days. A total of 126 patients were evaluable (received at least five doses of micafungin). Success (complete or partial response) was seen in 83.3% patients overall. Success rates for treatment of infections caused by the most common Candida spp. were as follows: C. albicans 85.1%, C. glabrata 93.8%, C. parapsilosis 86.4%, and C. tropicalis 83.3%. Serious adverse events related to micafungin were uncommon. Micafungin shows promise as a safe and effective agent for the treatment of newly diagnosed and refractory cases of candidemia. Large scale, randomized, controlled trials are warranted. PMID- 16261307 TI - Neurotoxicity induced by beta-lactam antibiotics: from bench to bedside. AB - Central nervous system toxicity following administration of beta-lactam antibiotics, of which penicillin is the prototype, is a potential cause of morbidity and mortality. In recent years, important advances have been made in the pathogenesis of antibiotic-related neurotoxicity. This review focuses on the experimental and clinical aspects of neurotoxicity caused by beta-lactam antibiotics. The purpose is to provide an update on the pathogenesis, mechanism, and clinical manifestations of the neurotoxicity, along with an overview of the relationship between antibiotic structure and convulsive action. In particular, some of the prevailing ideas about pathogenesis are highlighted, including theories of the mechanism of pathogenicity. A better understanding of antibiotic related neurotoxicity, as derived from animal models and human clinical experience, would be of value in facilitating more efficient and safer use of antimicrobial compounds. PMID- 16261308 TI - Impaired beta cell glucose sensitivity and whole-body insulin sensitivity as predictors of hyperglycaemia in non-diabetic subjects. AB - AIMS/HYPOTHESIS: The aim of this prospective study was to investigate predictors of deteriorating glucose tolerance in subjects of British extraction. METHODS: A total of 156 non-diabetic subjects (86 with a family history of type 2 diabetes) underwent a 75-g OGTT and anthropometric assessment at baseline and 5 years later. Pancreatic beta cell function and whole-body insulin sensitivity were studied by model assessment. Subjects were classified as progressors if glucose tolerance moved one or more steps from normal, impaired fasting glucose, impaired glucose tolerance and diabetes over the follow-up period. RESULTS: At baseline, the progressors (n=22) had increased adiposity and a higher proportion of familial diabetes and abnormal glucose tolerance than non-progressors. Baseline pancreatic beta cell sensitivity to changes in glucose (p<0.02) and whole-body insulin sensitivity (p<0.0001) were decreased in the progressors. Logistic regression revealed that baseline and follow-up changes in beta cell glucose sensitivity and insulin sensitivity, rather than the classical clinical predictors (adiposity, familial diabetes and glucose levels), were the key independent predictors of progression (explaining over 50% of the progression). CONCLUSIONS/INTERPRETATION: Impaired pancreatic beta cell glucose sensing and whole-body insulin sensitivity predict progression to hyperglycaemia. Strikingly, these pathophysiological changes override the importance of the clinical risk factors and highlight potential metabolic targets for prevention strategies. PMID- 16261309 TI - Renal hyperfiltration in type 2 diabetes: effect of age-related decline in glomerular filtration rate. AB - AIMS/HYPOTHESIS: We sought to characterise the effect of the age-related decline of GFR on hyperfiltration in type 2 diabetes and to identify clinical characteristics associated with hyperfiltration. MATERIALS AND METHODS: GFR was measured in 662 type 2 diabetic patients by plasma disappearance of 99 m technetium-diethylene-triamine-penta-acetic acid. The prevalence of hyperfiltration was calculated using both an age-unadjusted GFR threshold of >130 ml min(-1) 1.73 m(-2) and an age-adjusted threshold incorporating a decline of 1 ml min(-1) year(-1) after the age of 40. The hyperfiltering patients were compared with type 2 diabetic subjects who had a GFR between 90 and 130 ml min( 1) 1.73 m(-2) and were matched for age, sex and disease duration to allow for identification of modifiable factors associated with hyperfiltration. RESULTS: The prevalence of hyperfiltration was 7.4% when age-unadjusted and 16.6% when age adjusted definitions were used. The age-unadjusted vs -adjusted prevalence rates for hyperfiltration were 50 vs 50%, 12.9 vs 23.4% and 0.3 vs 9.0% for patients aged <40 years, 40 to 65 years and >65 years, respectively. Both the age unadjusted and -adjusted hyperfiltration groups had lower mean diastolic blood pressure and lower serum creatinine levels than the control groups. Although the age-unadjusted hyperfiltration group had larger kidneys compared to the control group, this difference was no longer significant when the age-adjusted definition was used. There were no differences in HbA(1)c, mean arterial pressure, antihypertensive use, insulin therapy, dyslipidaemia, frequency of macro- or microvascular complications, BMI, urinary sodium, urea and albumin excretion between the groups. CONCLUSIONS/INTERPRETATION: Hyperfiltration was still more common among younger patients with type 2 diabetes even after adjusting for the expected age-related decline in GFR. Hyperfiltration was associated with a lower mean diastolic blood pressure independent of age. PMID- 16261310 TI - The impact of ethnicity and sex on subclinical cardiovascular disease: the Diabetes Heart Study. AB - AIMS/HYPOTHESIS: African-Americans with type 2 diabetes and access to adequate healthcare are at lower risk of clinical coronary artery disease than are white diabetic patients. We evaluated whether ethnic differences in subclinical cardiovascular disease, coronary and carotid artery calcified plaque and carotid artery intima-medial thickness (IMT) were present in members of The Diabetes Heart Study families. SUBJECTS AND METHODS: In a bi-racial cohort of 1,180 individuals from families enriched for members with type 2 diabetes, we calculated coronary and carotid artery calcified plaque using fast-gated helical computed tomography, and measured carotid artery IMT and clinical risk factor profiles. Generalised estimating equations were used to test for an association between measures of subclinical cardiovascular disease and ethnicity and sex. RESULTS: After adjustment for age, ethnicity and kidney function, African Americans had significantly lower amounts of coronary artery calcified plaque (mean+/-SE) (866+/-158 vs 1,915+/-135, respectively; p=0.0466) and carotid artery calcified plaque (179+/-51 vs 355+/-27, respectively; p=0.0240) relative to whites, despite having increased carotid IMT (0.71+/-0.01 vs 0.67+/-0.004 cm, respectively; p=0.0007), and higher blood pressure, albuminuria and HbA1c. Sex specific analyses revealed that African-American men had significantly lower coronary and carotid artery calcified atheroma than white men. In women, ethnic differences in calcified carotid artery plaque, but not coronary artery plaque, were observed. CONCLUSIONS/INTERPRETATION: In families enriched for members with type 2 diabetes, African-American men had markedly lower levels of coronary and carotid artery calcified plaque than white men, despite increased carotid artery IMT and conventional risk factors. These findings suggest that susceptibility to subclinical cardiovascular disease differs markedly according to ethnicity and sex. PMID- 16261311 TI - Leptin : adiponectin ratio as an atherosclerotic index in patients with type 2 diabetes : relationship of the index to carotid intima-media thickness. PMID- 16261313 TI - Visual fields correlate better than visual acuity to severity of diabetic retinopathy. AB - AIMS/HYPOTHESIS: We compared the outcomes of perimetric and visual acuity tests in patients with diabetic retinopathy. METHODS: We examined 59 diabetic patients with different degrees of retinopathy using stereo fundus photography in accordance with the Early Treatment of Diabetic Retinopathy Study (ETDRS) and fluorescein angiography. Conventional white-on-white perimetry (WWP) and short wavelength automated perimetry (SWAP) were performed and analysed with reference to normal values. Visual acuity was measured with ETDRS charts. RESULTS: Regression analysis revealed that visual acuity was significantly associated with increasing severity of retinopathy according to the ETDRS scale when visual acuity was estimated by counting logarithm of minimum angle of resolution (LogMar) scores, but not when visual acuity was measured by the conventional reading of the smallest line that could be seen. Visual acuity decreased by 0.02 LogMar per ETDRS step (p=0.03). The degree of visual field loss was significantly associated with increasing severity of retinopathy according to the ETDRS scale, perimetric sensitivity decreasing by 0.44 dB per ETDRS step (p=0.0001) using WWP, and by 0.40 dB per ETDRS step (p=0.04) with SWAP. The size of the area of the foveal avascular zone and adjacent perifoveal intercapillary areas (PIAs) also affected the central visual field as obtained both by WWP (-2.6 dB/mm2, p=0.03), and by SWAP (-7.9 dB/mm2, p=0.002), but did not affect visual acuity. The regression model fit for peripheral retinopathy according to the ETDRS scale was better using WWP than SWAP or visual acuity, while SWAP testing was superior to both WWP and visual acuity when measuring effects caused by enlarged foveal avascular zones and PIAs. CONCLUSIONS/INTERPRETATION: Perimetry can provide more useful information than visual acuity on functional loss in diabetic retinopathy, particularly when the perifoveal capillary network is damaged. PMID- 16261312 TI - Reduced intrahepatic fat content is associated with increased whole-body lipid oxidation in patients with type 1 diabetes. AB - AIMS/HYPOTHESIS: Insulin resistance may be associated with ectopic fat accumulation potentially determined by reduced lipid oxidation. In patients with type 1 diabetes peripheral insulin resistance is associated with higher intramyocellular lipid content. We assessed whether these patients are also characterised by intrahepatic fat accumulation and abnormal fat oxidation. METHODS: Nineteen patients with type 1 diabetes (6 women, 13 men, age 35+/-7 years, BMI 23+/-3 kg/m2), HbA1c 8.7+/-1.4%) and 19 healthy matched individuals were studied by (1) euglycaemic-hyperinsulinaemic clamp combined with [6,6 2H2]glucose infusion to assess whole-body glucose metabolism; (2) indirect calorimetry to assess glucose and lipid oxidation; and (3) localised 1H-magnetic resonance spectroscopy of the liver to assess intrahepatic fat content. RESULTS: Patients with type 1 diabetes showed a reduced insulin-stimulated metabolic clearance rate of glucose (4.3+/-1.3 ml kg(-1) min(-1)) in comparison with normal subjects (6.0+/-1.6 ml kg(-1) min(-1); p<0.001). Endogenous glucose production was higher in diabetic patients (p=0.001) and its suppression was impaired during insulin administration (66+/-30 vs 92+/-8%; p=0.047) in comparison with normal subjects. Plasma glucagon concentrations were not different between groups. The estimated hepatic insulin concentration was lower in diabetic patients than in normal subjects (p<0.05), as was the intrahepatic fat content (1.5+/-0.7% and 2.2+/-1.0% respectively; p<0.03), the latter in association with a reduced respiratory quotient (0.74+/-0.05 vs 0.84+/-0.06; p=0.01) and increased fasting lipid oxidation (1.5+/-0.5 vs 0.8+/-0.4 mg kg(-1) min(-1); p<0.01). CONCLUSIONS/INTERPRETATION: In patients with type 1 diabetes, insulin resistance was not associated with increased intrahepatic fat accumulation. In fact, diabetic patients had reduced intrahepatic fat content, which was associated with increased fasting lipid oxidation. The unbalanced hepatic glucagon and insulin concentrations affecting patients with type 1 diabetes may be involved in this abnormality of intrahepatic lipid metabolism. PMID- 16261314 TI - Control of the phytopathogen Botrytis cinerea using adipic acid monoethyl ester. AB - The in vitro and in vivo antifungal activity of adipic acid monoethyl ester (AAME) on the necrotrophic pathogen Botrytis cinerea has been studied. This chemical effectively controlled this important phytopathogen, inhibited spore germination and mycelium development at non-phytotoxic concentrations. The effectiveness of AAME treatment is concentration-dependent and influenced by pH. Spore germination in the presence of AAME is stopped at a very early stage, preventing germ tube development. In addition, cytological changes such as retraction of the conidial cytoplasm in the fungus are observed. AAME was also found to act on membrane integrity, affecting permeability without exhibiting lytic activity, as described previously for other antifungal compounds. Polyamine content in the mycelium of B. cinerea was also affected in response to AAME treatment, resulting in putrescine reduction and spermine accumulation similar to a number of antifungal agents. Microscopic observation of treated conidia after inoculation on tomato leaves suggested that inhibited spores are not able to attach to and penetrate the leaf. Finally, AAME completely suppressed the grey mould disease of tomato fruits under controlled inoculation conditions, providing evidence for its efficacy in a biological context and for the potential use of this chemical as an alternative fungicide treatment. PMID- 16261315 TI - Effect of the cannabinoid CB1 receptor antagonist SR-141716A on ethanol self administration and ethanol-seeking behaviour in rats. AB - RATIONALE: It has been suggested that endocannabinoid mechanisms are involved in the control of ethanol consumption. OBJECTIVES: The aims of the present study were (1) to evaluate the role of the endocannabinoid system in the control of operant ethanol self-administration and in the reinstatement of ethanol seeking, when induced by stress or conditioned stimuli and (2) to offer new insights on the specificity of such a role. METHODS: Rats were administered intraperitoneally with the selective cannabinoid CB1 receptor antagonist, SR-141716A, 30 min before operant self-administration or reinstatement sessions. Two schedules of reinforcement, the fixed-ratio 1 (FR1) and the progressive ratio (PR), were used to study 10% (w/v) alcohol and 5.0% sucrose self-administration. NaCl (2% w/v) intake in sodium-depleted rats was studied only under the FR1 program. RESULTS: Treatment with SR-141716A (0.3-3.0 mg/kg) significantly attenuated FR1 alcohol self-administration and lowered the break point for ethanol under PR. SR-141716A also markedly inhibited the reinstatement of alcohol seeking elicited by presentation of cues predictive of drug availability. Conversely, the cannabinoid antagonist did not prevent the reinstatement of alcohol seeking induced by foot shock stress. Lever pressing for sucrose under FR1 and PR schedules was also significantly decreased by SR-141716A treatment, whereas the drug modestly and only at the highest dose decreased 2% NaCl self-administration. CONCLUSIONS: Results emphasize that endocannabinoid mechanisms play a major role in the control of ethanol self-administration and in the reinstatement of conditioned ethanol seeking. However, these effects extend to the control of operant behaviours motivated by natural rewards (i.e. sucrose). On the other hand, SR 141716A only weakly reduces NaCl self-administration in sodium-depleted rats, in which salt intake is largely controlled by homeostatic mechanisms. Overall, these observations demonstrate that the inhibition of operant behaviour following blockade of CB1 receptors by SR-141716A is linked to a reduction of reward related responding and is not related to drug-induced motor deficits. PMID- 16261316 TI - Contribution of serotonin (5-HT) 5-HT2 receptor subtypes to the discriminative stimulus effects of cocaine in rats. AB - RATIONALE: The serotonin (5-hydroxytryptamine; 5-HT) 5-HT2 receptor (5-HT2R) family is an important regulator of the behavioral responsiveness to cocaine. OBJECTIVE: The present study is an analysis of the role of the 5-HT2R subtypes (5 HT2AR, 5-HT2BR, and 5-HT2CR) in the discriminative stimulus effects of cocaine. METHODS: Male Wistar rats were trained to discriminate cocaine (10 mg/kg) from saline in a two-lever, water-reinforced FR 20 task, and we investigated the ability of the 5-HT2AR antagonist 1(Z)-[2-(dimethylamino)ethoxyimino]-1(2 fluorophenyl)-3-(4-hydroxyphenyl)-2(E)-propene (SR 46349B), the 5-HT2BR antagonist N-(1-methyl-5-indolyl)-N'-(3-methyl-5-isothiazolyl) urea (SB 204741), and the 5-HT2CR antagonist [(+)-cis-4,5,7a,8,9,10,11,11a-octahydro-7H-10 methylindolo(1,7-bC)(2,6)naphthyridine (SDZ SER-082) to substitute for or to modulate the stimulus effects of cocaine. RESULTS: Pretreatment with SR 46349B (0.5-1 mg/kg) resulted in a rightward shift of the cocaine dose-response curve, while SDZ SER-082 (1 mg/kg) shifted the dose-response for cocaine to the left; SB 204741 (1-3 mg/kg) was inactive. CONCLUSIONS: Our pharmacological analyses of selective antagonists of 5-HT2AR, 5-HT2BR, and 5-HT2CR indicate oppositional influence of 5-HT2AR and 5-HT2CR on the stimulus effects of cocaine and exclude a role for the 5-HT2BR. These data suggest that 5-HT2AR and 5-HT2CR may be important in modulating the subjective effects of cocaine in humans. PMID- 16261317 TI - Reduction of abstinence-induced withdrawal and craving using high-dose nicotine replacement therapy. AB - RATIONALE: Decreasing withdrawal and craving during smoking cessation is a major aim of cessation medications. Prior studies have shown that Nicotine Replacement Therapy (NRT) decreases withdrawal symptom severity but have relied on retrospective reports and lacked robust measures of baseline symptoms or symptoms during unmedicated abstinence. OBJECTIVES AND METHODS: We tested the effect of high-dose (35 mg) nicotine patch on withdrawal and craving during abstinence using real-time assessment with electronic diaries during ad libitum smoking, a brief period of experimentally directed trial abstinence, and the first 3 days of cessation. Subjects were 324 smokers randomized to high-dose nicotine patches or placebo. RESULTS: Treatment with active patches reduced withdrawal and craving during cessation and completely eliminated deprivation-related changes in affect or concentration. CONCLUSION: High-dose NRT reduces withdrawal symptoms and craving and can eliminate some symptoms entirely. PMID- 16261319 TI - Fluorometric determination of thiol redox state. AB - This work presents a fluorometric thiol redox state (TRS) method based on the thiol-specific fluorescent probe monobromobimane. It determines the concentrations of non-protein (NP) thiols glutathione, cysteine and N acetylcysteine, the protein (P) thiols, as well as the contribution of these components to symmetric and mixed disulfides (NPSSR, NPSSC, NPSSCAc, PSSR, PSSC, PSSCAc, PSSP). The method is very sensitive since it measures as low as 30 pmol SH groups in samples with a minimum of 1-5 mg total protein, making possible the measurement of oxidative stress-related TRS components even in biological fluids such as cerebrospinal. PMID- 16261320 TI - Feasibility of analyzing molecular pigments in paint layers using TOF S-SIMS. AB - A first attempt to measure the molecular compositions of pigments in paintings using static SIMS was made. An investigation of pellets of pure pigments such as auripigment and verdigris allowed the detection of numerous high m/z ions useful for molecular identification. Analysis of pigments in embedded paint fragments, on the other hand, only yielded elemental information because of charge build-up and contamination problems. Optimization of the form in which the sample is presented to the analysis method is obviously the price to pay for the ultimate sensitivity and information depth of S-SIMS. PMID- 16261318 TI - Surface-enhanced Raman scattering detection of lysophosphatidic acid. AB - Surface-enhanced Raman scattering using silver nanoparticles was applied to detect various forms of lysophosphatidic acid (LPA) to examine its potential application as an alternative to current detection methods of LPA as biomarkers of ovarian cancer. Enhancement of the Raman modes of the molecule, especially those related to the acyl chain within the 800-1300 cm(-1) region, was observed. In particular, the C-C vibration mode of the gauche-bonded chain around 1100 cm( 1) was enhanced to allow the discrimination of two similar LPA molecules. Given the molecular selectivity of this technique, the detection of LPA using SERS may eliminate the need for partial purification of samples prior to analysis in cancer screening. PMID- 16261321 TI - Solution to the mass spectrometry challenge. PMID- 16261322 TI - Spectroscopy challenge 10. PMID- 16261323 TI - Analysis of alkyl polyglucosides in industrial products by capillary electrophoresis with pulsed amperometric detection. AB - Pulsed amperometric detection following micellar electrokinetic chromatography has been applied successfully to the direct detection of alkyl polyglucosides (APGs) in shampoos and other industrial products without prior conversion to highly absorbing or fluorescing derivatives. For electrochemical detection, it is necessary to dissociate the hydroxyl groups of the APGs. Thus, we used 0.1 M NaOH in the outlet vial to dissociate the APGs. The main problems associated with the combination of electrochemical detection and capillary electrophoresis are the need to isolate the detector from the electric field used in the capillary electrophoresis separation and the difficulty of aligning the working electrode with the end of the capillary. To overcome these problems, a simple capillary electrode holder was constructed. This holder automatically aligns the capillary and the electrode in a wall-jet configuration without the aid of micropositioners and facilitates the replacement of electrodes and capillaries without reconstruction of the entire capillary/electrode setup. Special microcylindrical gold electrodes have been produced by sealing 300-microm-diameter gold wire into borosilicate-glass capillaries. PMID- 16261326 TI - Aquatic degradation of triclosan and formation of toxic chlorophenols in presence of low concentrations of free chlorine. AB - The degradation of 2-(2,4-dichlorophenoxy)-5-chlorophenol (triclosan) in chlorinated water samples was investigated. Sensitive determination of the parent compound and its transformation products was achieved by gas chromatography with mass spectrometry detection after sample concentration, using a solid-phase extraction sorbent and silylation of the target compounds. Experiments were accomplished using ultrapure water spiked with chlorine and triclosan concentrations in the low mg/l and ng/ml ranges respectively. Chlorination of the phenolic ring and cleavage of the ether bond were identified as the main triclosan degradation pathways. Both processes led to the production of two tetra and a penta-chlorinated hydroxylated diphenyl ether, as well as 2,4 dichlorophenol. The formation of 2,3,4-trichlorophenol was not detected in any experiment; however, significant amounts of 2,4,6-trichlorophenol were noticed. All of these five compounds were also identified when triclosan was added to tap water samples with free chlorine concentrations below 1 mg/l. Minor amounts of three di-hydroxylated phenols, containing from one to three atoms of chlorine in their structures, were also identified as unstable triclosan chlorination by products. The analysis of several raw wastewater samples showed the co-existence of important concentrations of triclosan and its most stable by-products (2,4 dichlorophenol and 2,4,6-trichlorophenol), reinforcing the potential occurrence of the described transformations when products containing triclosan are mixed with chlorinated tap water. PMID- 16261327 TI - Detection of testosterone, nandrolone and precursors in horse hair. AB - Growing interest among several horse-breeder associations has initiated the development of a screening procedure to test for anabolic agents in hair, which has the advantage over blood and urine specimens of allowing long-term detection. An analytical method was established to monitor in tails or manes several anabolic substances available as veterinary medicines or as so-called nutritional supplements (clenbuterol, different esters or prohormones of nandrolone and testosterone). The analytical procedure to detect steroids in hair samples consists of the following steps: decontamination of the hair strand or segment with methanol/water (1:1), milling, extraction of the hair material in an ultrasonic bath using methanol, purification by liquid-liquid extraction (n pentane/methanol, 25:1) and HPLC cleanup, derivatisation of the relevant LC fractions with MSTFA, and measurement using GC-MS/MS technique. The first objective of our study was the detection of exogenous nandrolone (nortestosterone, NT) in the horse hair; therefore nandrolone-associated compounds [nandrolone dodecanoate administered intramuscularly (i.m.) and a mixture of 4-estrenediol and 4-estrenedione, transdermal] were administered to four geldings. The highest concentrations of NT following i.m. treatment were measured after 10 days in a 2-cm hair segment (up to 18 pg/mg); NT was detectable for up to 120 days and in some cases up to 330 days in tail hair (limit of detection 0.3 pg/mg). Following transdermal application, nandrolone as well as the administered prohormones were identified in tail and mane until the latest sampling at 3 months. Furthermore, untreated stallions (128) were investigated to estimate the range of endogenous levels of NT and testosterone (T) in hair. Maximum values of 3 pg/mg (NT) and 1 pg/mg (T) were quantified originating from endogenous formation in the male horse. Additionally, a possible relationship between steroid concentrations in hair specimens and the age of stallions was appraised. NT and T were not detected in hair samples of control geldings. Following nandrolone treatment of geldings, highest values in hair exceeded the endogenous amount detected in untreated stallions. Therefore comparison of concentrations measured in control samples with the estimated endogenous levels could give a clue to exogenous application in cases of abnormally high amounts of NT or T. The possibility of the evaluation of threshold values is discussed as a means to verify an exogenous administration of NT and T in hair samples. Furthermore, the detection of a synthetic substance in hair, e. g. the parent steroid ester by itself, would be unequivocal proof of an exogenous origin of NT or T and the previous medication of the stallion. PMID- 16261328 TI - Histological evidence for the role of mechanical stress in modulating thermal denaturation of collagen. AB - The hyperthermia and thermal denaturation literatures reveal a time-temperature equivalency when heating cells or connective tissues: thermal damage increases with increasing temperature (for the same duration) and increases with increasing duration (for the same temperature). Recent findings conversely suggest that increasing the mechanical loading on a tissue during heating decreases the thermal damage (for a given temperature and duration of heating). Surprisingly, however, there are few histological correlates of such damage. In this paper, we show that progressive light microscopic changes - swelling of collagen bands, thickening of collagen-rich layers, hyalinization, and loss of birefringence approximately - correlate very well with both increased heating times and decreased mechanical loading. Increased mechanical stress is thus thermally protective and should be considered in the design of clinical procedures that use heating to treat diseases or injuries. PMID- 16261329 TI - Exploitation of a marker dense linkage map of potato for positional cloning of a wart disease resistance gene. AB - A marker-saturated linkage map of potato was used to genetically map a locus involved in the resistance against wart disease Synchytrium endobioticum race 1. The locus mapped on the long arm of chromosome 4 and is named Sen1-4 in contrast to a Sen1 locus on chromosome 11. The AFLP markers from the Sen1-4 interval enabled the isolation of BAC clones from an 11 genome equivalent BAC library. This was achieved via fingerprinting of BAC pools with the AFLP primer pairs that resemble the genetic marker loci. With non-selective AFLP primers, fingerprints of individual BAC clones were generated to analyse the overlap between BAC clones using FPC. This resulted in a complete contig and a minimal tiling path of 14 BAC clones enclosing the Sen1-4 locus. The BAC contig has a genetic length of approximately 6 cM and a physical length of approximately 1 Mb. Our results demonstrate that map-based cloning of Sen1-4 can be pursued on the basis of a strategy of marker saturation alone. Genetic resolution achieved by screening large numbers of offspring for recombination events may not be required. Together with the construction of the BAC contig, a physical map with the position of the markers is accomplished in one step. This provides proof of concept for the utility of the marker saturation that is offered by the ultra dense AFLP map of potato for gene cloning. PMID- 16261330 TI - Accelerated myelination associated with venous congestion. AB - Magnetic resonance imaging is currently the gold standard in the assessment of brain myelination. The normal pattern of brain myelination conforms to a fixed chronological sequence. Focal accelerated myelination is a usual pathological state and previously has only been associated with Sturge-Weber syndrome. The purpose of our study is to describe alternate causes for accelerated myelination. We retrospectively reviewed serial MR scans, MR angiography, conventional angiography and the clinical progress of three children with accelerated myelination. Two patients with accelerated myelination had an underlying cerebral sinovenous thrombosis. The third patient had Sturge-Weber syndrome. Our study strongly suggests that cerebral venous thrombosis with the consequent restriction of venous outflow could be a key factor in the induction of accelerated myelination. We recommend that in patients with accelerated myelination, the search for an underlying etiology should include careful evaluation of the intracranial vascular pathology, especially cerebral venous thrombosis. PMID- 16261332 TI - Spine anatomy reveals the diversity of catfish through time: a case study of Synodontis (Siluriformes). AB - Synodontis (Mochokidae, Siluriformes) is a freshwater catfish endemic to Africa. The 118 extant species are present in almost all hydrographic basins. Some species are restricted to a single stream, whereas others have a vast distribution. Synodontis is known in the fossil record since the Miocene, and its history depends on the connections among African basins through time. The identification of species in the fossil record is essential to reconstruct this historical pattern. Catfish pectoral and dorsal spines are robust, they preserve well and they form most of the fossil remains for the genus Synodontis. Unfortunately, the criteria for the identification of extant Synodontis species are not applicable to fossil specimens. Here, we define 11 original morphological characters that permit to discriminate four extant species from the Chad-Chari hydrographic system. Six of these characters are defined on pectoral spines and five on dorsal spines. We then show that these characters can be used successfully for identifying fossil specimens. In particular, we present a case study in which we identify Synodontis cf. schall and Brachysynodontis cf. batensoda in the hominid-bearing sector Toros-Menalla (Late Miocene, northern Chad). We show that spine anatomy can be a powerful tool to recognise catfish species through time and thus to identify historical diversity pattern. PMID- 16261334 TI - Imaging features of copper deficiency myelopathy: a study of 25 cases. AB - Acquired copper deficiency presents with a spastic gait and sensory ataxia. Spinal cord magnetic resonance imaging (MRI) in patients with copper deficiency myelopathy may show increased T2 signal, most commonly in the dorsal midline cervical and thoracic cord. These imaging findings may be reversible with normalization of serum copper. The clinical and imaging picture is very similar to the subacute combined degeneration seen in patients with vitamin B12 deficiency. Neuroradiologists should consider this possibility when a long segment of symmetric dorsal spinal cord T2-hyperintensity is identified. PMID- 16261335 TI - Intraarterial injection of colforsin daropate hydrochloride for the treatment of vasospasm after aneurysmal subarachnoid hemorrhage: preliminary report of two cases. AB - We describe two patients with symptomatic vasospasms after aneurysmal subarachnoid hemorrhage who were successfully treated with intraarterial injection of colforsin daropate hydrochloride (HCl). Colforsin daropate HCl is capable of directly stimulating adenylate cyclase, which in turn causes vasorelaxation via elevated intracellular concentrations of cyclic adenosine monophosphate. We suggest that colforsin daropate HCl might be a useful therapeutic tool in treating cerebral vasospasm. PMID- 16261331 TI - Nuclear presence of adhesion-/growth-regulatory galectins in normal/malignant cells of squamous epithelial origin. AB - Cellular activities in the regulation of growth or adhesion/migration involve protein (lectin)-carbohydrate recognition at the cell surface. Members of the galectin family of endogenous lectins additionally bind distinct intracellular ligands. These interactions with protein targets explain the relevance of their nuclear and cytoplasmic presence. Expression profiling for galectins and accessible binding sites is a histochemical approach to link localization with cellular growth properties. Non-cross-reactive antibodies for the homodimeric (proto-type) galectins-1, -2 and -7 and the chimera-type galectin-3 (Gal-3) as well as the biotinylated lectins were tested. This analysis was performed with the FaDu squamous carcinoma cell line and long-term cultured human and porcine epidermal cells as models for malignant and normal cells of squamous cell epithelial origin. A set of antibodies was added for phenotypic cell characterization. Strong nuclear and cytoplasmic signals of galectins and the differential reactivity of labeled galectins support the notion of their individual properties. The length of the period of culture was effective in modulating marker expression. Cytochemical expression profiling is a prerequisite for the selection of distinct proteins for targeted modulation of gene expression as a step toward functional analysis. PMID- 16261333 TI - Src homology 2-containing phosphotyrosine phosphatase regulates endothelin-1 induced epidermal growth factor receptor transactivation in rat renal tubular cell NRK-52E. AB - Epidermal growth factor (EGF) and endothelin-1 (ET-1) have been shown to be involved in proliferation and autoregeneration of renal tubular cells. This study aims to investigate the regulatory mechanism of ET-1-mediated EGF receptor (EGFR) transactivation in rat renal tubular cells (NRK-52E). Exposure of NRK-52E cells to ET-1 was found to stimulate the phosphorylation of EGFR and induce reactive oxygen species (ROS) generation. Both NAD(P)H oxidase inhibitor, diphenyliodonium (DPI) and ROS scavenger N-acetylcysteine (NAC), inhibited EGFR transactivation and extracellular signal-regulated kinase (ERK) phosphorylation caused by ET-1. In contrast, blockade of EGFR by AG1478 inhibited the phosphorylation of ERK but not ROS generation following ET-1 exposure. We found that the catalytic cysteine of Src homology 2-containing phosphotyrosine phosphatase (SHP-2) was transiently oxidized by ET-1 treatment in a modified malachite green phosphatase assay. In EGFR co-immunoprecipitation, SHP-2 was also found to interact with EGFR following ET-1 treatment. In SHP-2 knockdown NRK-52E cells, ET-1-induced EGFR transactivation was dramatically elevated and not influenced by NAC. However, GM6001 (an MMP inhibitor) and heparin binding (HB)-EGF neutralizing antibody suppressed this elevation. Our data suggest that ROS-mediated oxidation of SHP-2 is essential for HB-EGF-mediated EGFR transactivation in ET-1 signaling pathway in NRK-52E cells. PMID- 16261336 TI - An evaluation of immediate sheath removal and use of the Angio-Seal vascular closure device in neuroradiological interventions. AB - We set out to assess the effect of using an arteriotomy closure device in patients at high risk of access site complications. One-hundred nineteen patients with femoral artery access were prospectively included in five centers in France (56 males, 63 females, age range 20-84 years). The neurointerventional procedures were embolization of intracranial aneurysm (71.4%), carotid angioplasty (11.8%), cerebral angioplasty (1.7%), thrombolysis (0.8%), and other procedures (14.3%). Sheaths were removed and the closure device deployed immediately postprocedure. One patient developed a false aneurysm, and one developed a small hematoma. Neither required treatment. Postdevice deployment bleeding was experienced in three patients (2.5%), requiring minimal management. There were no major complications. Three patients died of causes unrelated to the vessel closure. Patient comfort was 2.2 on a scale of 1 (none) to 10 (severe). The results showed that immediate sheath removal followed by use of the closure device is safe in patients undergoing neurointerventional procedures and offers a low level of patient discomfort. In selected patients, early hospital discharge was possible. PMID- 16261337 TI - Kinematic and non-kinematic signals transmitted to the cat cerebellum during passive treadmill stepping. AB - Previous work from this laboratory has shown that activity in the dorsal spinocerebellar tract (DSCT) relates strongly to global hindlimb kinematics variables during passive displacements of the hindlimb. A linear relationship to limb axis orientation and length variables accounts for most of the response variance for passive limb positioning and movement. Here we extend those observations to more natural movements by examining the information carried by the DSCT during passive stepping movements on a treadmill, and we compare it to information transmitted during passive robot-driven hindlimb movements. Using a principal component analysis approach, we found that a linear relationship between the responses and hindlimb kinematics was comparable across experimental conditions. We also observed systematic non-linearities in this relationship for both types of movement that could be attributed to events corresponding to the touch-down and lift-off phases of the movement. We concluded that proprioceptive information transmitted to the cerebellum by the DSCT during locomotion has at least two major components. One component is associated with limb kinematics (limb orientation) and may be more or less related to the metrics of the step (stride length, for example) or its velocity. The other component is associated with limb length and/or limb loading, and it may signal some aspect of limb stiffness. PMID- 16261338 TI - Recruitment of gastrocnemius muscles during the swing phase of stepping following partial denervation of knee flexor muscles in the cat. AB - In walking cats, the biarticular medial and lateral gastrocnemius (MG-LG) muscles act to produce extension and flexion torques at the ankle and knee, respectively, and they usually display only one burst of activity beginning just before ground contact and ending near the end of the stance phase. Currently, the MG-LG muscles are considered to function primarily to control extension movements around the ankle joint during the stance phase. However, their flexion action at the knee means that they have the capacity to regulate rotations at the knee, but this role has not yet been clearly defined. Following partial denervation of the other muscles that normally act to flex the knee during swing, we observed that the MG LG muscles, but not the Soleus muscle (a pure ankle extensor), often generated strong bursts of activity during early swing. These bursts were enhanced following mechanical stimulation of the paw, and they were especially prominent when the leg trailed over an object. They were absent when the leg led over an object. During treadmill walking the swing-related bursts in MG and LG had little influence on ankle flexion at the beginning of swing, but they were associated with slowing of ankle flexion when the leg trailed over an object. We hypothesized that the recruitment of these bursts functions to partially compensate for the reduction in knee torque resulting from the denervation of other knee flexors. Consistent with this hypothesis was our finding that the magnitude of the swing-related activity in the MG-LG muscles was linearly correlated to the extent of the knee flexion and to the peak angular velocity of knee flexion, and that the timing of the bursts was similar to that in the denervated muscles prior to denervation. Our findings suggest that an excitatory pathway exists from the flexor half-center of the central pattern-generating network to MG-LG motoneurons, and that this pathway is strongly regulated by central and/or peripheral signals. PMID- 16261339 TI - Experimental treatment for focal hyperglycemic ischemic brain injury in the rat. AB - Hyperglycemia aggravates ischemic brain injury, possibly due to the activation of signaling pathways involving reactive oxygen species, Src and mitogen-activated protein kinases. The aim of this study was to investigate the effects of the spin trap agent alpha-phenyl-N-tert-butyl nitrone (PBN), the Src family kinase inhibitor PP2 and the MEK1-inhibitor U0126 on focal hyperglycemic ischemic brain injury. Temporary middle cerebral artery occlusion (90 min) was induced in four groups of rats (PBN, PP2, and U0126 vs. control). Neurological testing and tetrazolium red staining were performed after 1 day. PBN decreased the infarct volume by 70% compared with the control (P<0.05) and a tendency towards reduced infarcts was seen in the PP2 or U0126 groups. Furthermore, neurological testing was consistent with the volumetric analysis. In conclusion, PBN appears to be a potential neuroprotective agent in hyperglycemic, focal ischemic brain injury, while the efficacy of PP2 and U0126 could not be confirmed by the present data. PMID- 16261340 TI - Contribution of reference frames for movement planning in peripersonal space representation. AB - The principal goal of our study is to gain an insight into the representation of peripersonal space. Two different experiments were conducted in this study. In the first experiment, subjects were asked to represent principal anatomical reference planes by drawing ellipses in the sagittal, frontal and horizontal planes. The three-dimensional hand-drawing movements, which were achieved with and without visual guidance, were considered as the expression of a cognitive process per se: the peripersonal space representation for action. We measured errors in the spatial orientation of ellipses with regard to the requested reference planes. For ellipses drawn without visual guidance, with eyes open and eyes closed, orientation errors were related to the reference planes. Errors were minimal for sagittal and maximal for horizontal plane. These disparities in errors were considerably reduced when subjects drew using a visual guide. These findings imply that different planes are centrally represented, and are characterized, by different errors when subjects use a body-centered frame for performing the movement and suggest that the representation of peripersonal space may be anisotropic. However, this representation can be modified when subjects use an environment-centered reference frame to produce the movement. In the second experiment, subjects were instructed to represent, with eyes open and eyes closed, sagittal, frontal and horizontal planes by pointing to virtual targets located in these planes. Disparities in orientation errors measured for pointing were similar to those found for drawing, implying that the sensorimotor representation of reference planes was not constrained by the type of motor tasks. Moreover, arm postures measured at pointing endpoints and at comparable spatial locations in drawing are strongly correlated. These results suggest that similar patterns of errors and arm posture correlation, for drawing and pointing, can be the consequence of using a common space representation and reference frame. These findings are consistent with the assumption of an anisotropic action related representation of peripersonal space when the movement is performed in a body-centered frame. PMID- 16261341 TI - Changes in muscle tissue oxygenation during stagnant ischemia in septic patients. AB - OBJECTIVE: To determine changes in the rate of thenar muscles tissue deoxygenation during stagnant ischemia in patients with severe sepsis and septic shock. DESIGN AND SETTING: Prospective observational study in the medical ICU of a general hospital. PATIENTS AND PARTICIPANTS: Consecutive patients admitted to ICU with septic shock (n=6), severe sepsis (n=6), localized infection (n=3), and healthy volunteers (n=15). INTERVENTIONS: Upper limb ischemia was induced by rapid automatic pneumatic cuff inflation around upper arm. MEASUREMENTS AND RESULTS: Thenar muscle tissue oxygen saturation (StO2) was measured continuously by near-infrared spectroscopy before and during upper limb ischemia. StO(2) before intervention was comparable in patients with septic shock, severe sepsis, or localized infection and healthy volunteers (89 [65, 92]% vs. 82 [72, 91]% vs. 87 [85, 92]% vs. 83 [79, 93]%, respectively; p>0.1). The rate of StO(2) decrease during stagnant ischemia after initial hemodynamic stabilization was slower in septic shock patients than in those with severe sepsis or localized infection and in controls (-7.0 [-3.6, -11.0] %/min vs. -10.4 [-7.8, -13.3] %/min vs. -19.5 [ 12.3, -23.3] vs. -37.4 [-27.3, -56.2] %/min, respectively; p=0.041). At ICU discharge the rate of StO2 decrease did not differ between the septic shock, severe sepsis, and localized infection groups (-17.0 [-9.3, -28.9] %/min vs. 19.9 [-13.3, -23.6] %/min vs. -23.1 [-20.7, -26.2] %/min, respectively), but remained slower than in controls (p<0.01). The rate of StO2 decrease was correlated with Sequential Organ Failure Assessment (SOFA) score (r=0.739, p<0.001). CONCLUSIONS: After hemodynamic stabilization thenar muscle tissue oxygen saturation during stagnant ischemia decreases slower in septic shock patients than in patients with severe sepsis or localized infection and in healthy volunteers. During ICU stay and improvement of sepsis the muscle tissue deoxygenation rate increases in survivors of both septic shock and severe sepsis and was correlated with SOFA score. PMID- 16261342 TI - A novel autosomal recessive non-syndromic hearing impairment locus (DFNB47) maps to chromosome 2p25.1-p24.3. AB - Hereditary hearing impairment (HI) displays extensive genetic heterogeneity. Autosomal recessive (AR) forms of prelingual HI account for approximately 75% of cases with a genetic etiology. A novel AR non-syndromic HI locus (DFNB47) was mapped to chromosome 2p25.1-p24.3, in two distantly related Pakistani kindreds. Genome scan and fine mapping were carried out using microsatellite markers. Multipoint linkage analysis resulted in a maximum LOD score of 4.7 at markers D2S1400 and D2S262. The three-unit support interval was bounded by D2S330 and D2S131. The region of homozygosity was found within the three-unit support interval and flanked by markers D2S2952 and D2S131, which corresponds to 13.2 cM according to the Rutgers combined linkage-physical map. This region contains 5.3 Mb according to the sequence-based physical map. Three candidate genes, KCNF1, ID2 and ATP6V1C2 were sequenced, and were found to be negative for functional sequence variants. PMID- 16261344 TI - Expression pattern of Bombyx vasa-like (BmVLG) protein and its implications in germ cell development. AB - Germ cell development in the silkworm Bombyx mori is interesting in that the species has no recognizable germ plasm, and its germ cells appear first on the ventral side of the embryo, not on the posterior pole as in Drosophila melanogaster. We previously reported the isolation of a vasa homologue (BmVLG) from B. mori and revealed the specific expression of transcript in the germ cells. In this paper, we describe the embryonic expression pattern of BmVLG protein. Consistent with the lack of recognizable germ plasm, the protein is not localized in freshly laid eggs, and its specific expression is first detectable several hours after energids penetrate the periplasm. This is in contrast to D. melanogaster, where germ cell lineage can be traced with anti-vasa antibody just after the formation of pole cells as they sequester vasa-positive germ (pole) plasm during cellularization. It is also revealed that, within the first few hours of their appearance when extensive cell movement does not seem to occur, stained cells are sometimes widely dispersed along the midline, which eventually may lead to the formation of ectopic germ cells. The implications of these results for germ cell development are discussed. PMID- 16261343 TI - Contrasting patterns of Y-chromosome variation in South Siberian populations from Baikal and Altai-Sayan regions. AB - In order to investigate the genetic history of autochthonous South Siberian populations and to estimate the contribution of distinct patrilineages to their gene pools, we have analyzed 17 Y-chromosomal binary markers (YAP, RPS4Y(711), SRY-8299, M89, M201, M52, M170, 12f2, M9, M20, 92R7, SRY-1532, DYS199, M173, M17, Tat, and LLY22 g) in a total sample of 1,358 males from 14 ethnic groups of Siberia (Altaians-Kizhi, Teleuts, Shors, Tuvinians, Todjins, Tofalars, Sojots, Khakassians, Buryats, Evenks), Central/Eastern Asia (Mongolians and Koreans) and Eastern Europe (Kalmyks and Russians). Based on both, the distribution pattern of Y-chromosomal haplogroups and results on AMOVA analysis we observed the statistically significant genetic differentiation between the populations of Baikal and Altai-Sayan regions. We suggest that these regional differences can be best explained by different contribution of Central/Eastern Asian and Eastern European paternal lineages into gene pools of modern South Siberians. The population of the Baikal region demonstrates the prevalence of Central/Eastern Asian lineages, whereas in the populations of Altai and Sayan regions the highest paternal contribution resulted from Eastern European descent is revealed. Yet, our data on Y-chromosome STRs variation demonstrate the clear differences between the South Siberian and Eastern European R1a1-lineages with the evolutionary ages compatible with divergence time between these two regional groups. PMID- 16261345 TI - Dominant expression of 85-kDa form of cortactin in colorectal cancer. AB - PURPOSE: Cortactin is commonly expressed in several human cancers, which may alter their invasive or metastatic properties. Eighty five kilodalton form (p85) and 80-kDa form (p80) of cortactin are two separate bands in SDS-PAGE representing different conformational states. The objective of this study was to investigate cortactin expression in colorectal cancer (CRC). EXPERIMENTAL DESIGN: Cortactin expression was studied in an eight paired laser capture microdissection (LCM) CRC tissues and matched non-cancerous epithelia by immunoblotting. The expression in 58 CRC and two cell lines, HCT8 and HCT116, was studied respectively by immunohistochemistry and confocal laser scanning immunofluorescence. RESULTS: Dominant expression of p85 was identified in LCM procured CRC tissues compared with equal intensity of p85 and p80 forms in non cancerous tissues, while the amount of total cortactin was approximate. Immunohistochemistry analysis demonstrated that cortactin located in the cytoplasm of tumor cells and adjacent non-cancerous cells, and its expression was negatively correlated with TNM staging and lymphatic invasion status. However, the invasion fronts in 3 of 58 primary tumors and 28 of 39 available lymph node metastases were intensively stained. Further, immunofluorescence analysis showed that cortactin was distributed in cytoplasm and enriched in the front of the extending lamellipodia at adhering side of cultured cancer cells. CONCLUSIONS: Our results demonstrated the dominant expression of p85 form of cortactin in CRC for the first time. The enrichment of cortactin in the invasion front of some tumor cells and in the extending lamellipodia of cultured cancer cells suggests that cortactin may help cancer cell movement. PMID- 16261347 TI - Fine mapping of inherent flexibility variation along DNA molecules: validation by atomic force microscopy (AFM) in buffer. AB - Curvature and flexibility are structural properties of central importance to genome function. However, due to the difficulties in finding suitable experimental conditions, methods for studying one without the interference of the other have proven to be difficult. We propose a new approach that provides a measure of inherent flexibility of DNA by taking advantage of two powerful techniques, X-ray crystallography and nuclear magnetic resonance. Both techniques are able to detect local curvature on DNA fragments but, while the first analyzes DNA in the solid state, the second works on DNA in solution. Comparison of the two data sets allowed us to calculate the relative contribution to flexibility of the three rotations and three translations, which relate successive base pair planes for the ten different dinucleotide steps. These values were then used to compute the variation of flexibility along a given nucleotide sequence. This allowed us to validate the method experimentally through comparisons with maps of local fluctuations in DNA molecule trajectory constructed from atomic force microscopy imaging in solution. We conclude that the six dinucleotide-step parameters defined here provide a powerful tool for the exploration of DNA structure and, consequently will make an important contribution to our understanding of DNA-sequence-dependent biological processes. PMID- 16261348 TI - A homozygous recA mutant of Synechocystis PCC6803: construction strategy and characteristics eliciting a novel RecA independent UVC resistance in dark. AB - We report here the construction of a homozygous recA460::cam insertion mutant of Synechocystis sp. PCC 6803 that may be useful for plant molecular genetics by providing a plant like host free of interference from homologous recombination. The homozygous recA460::cam mutant is highly sensitive to UVC under both photoreactivating and non-photoreactivating conditions compared to the wild type (WT). The liquid culture of the mutant growing in approximately 800 lx accumulates nonviable cells to the tune of 86% as estimated by colony counts on plates incubated at the same temperature and light intensity. The generation time of recA mutant in standard light intensity (2,500 lx) increases to 50 h compared to 28 h in lower light intensity (approximately 800 lx) that was used for selection, thus explaining the earlier failures to obtain a homozygous recA mutant. The WT, in contrast, grows at faster rate (23 h generation time) in standard light intensity compared to that at approximately 800 lx (26 h). The Synechocystis RecA protein supports homologous recombination during conjugation in recA (-) mutant of Escherichia coli, but not the SOS response as measured by UV sensitivity. It is suggested that using this homozygous recA460::cam mutant, investigations can now be extended to dissect the network of DNA repair pathways involved in housekeeping activities that may be more active in cyanobacteria than in heterotrophs. Using this mutant for the first time we provide a genetic evidence of a mechanism independent of RecA that causes enhanced UVC resistance on light to dark transition. PMID- 16261346 TI - Proteomic analysis on metastasis-associated proteins of human hepatocellular carcinoma tissues. AB - PURPOSE: A comparative proteomic approach was used to identify and analyze proteins related to metastasis of hepatocellular carcinoma (HCC). METHODS: Proteins extracted from 12 HCC tissue specimens (six with metastases and six without) were separated by two-dimensional gel electrophoresis (2-DE). The protein spots exhibiting statistical alternations between the two groups through computerized image analysis were then identified by mass spectrometry. In addition immunohistochemistry (IHC), Western blotting and RT-PCR were performed to verify the expression of certain candidate proteins. RESULTS: 16 proteins including HSP27, S100A11, CK18 were annotated by mass spectrometry, relevant to chaperone function, cell mobility, cytoskeletal architecture, respectively. Most were previously unconnected with metastasis of HCC. Of these HSP27 was found overexpressed consistently in 2-DE patterns of all metastatic HCC tissues compared with nonmetastatic ones. IHC and Western blotting of HCC tissues confirmed this difference while RT-PCR did not. CONCLUSION: There are various proteins joined together in HCC metastasis. The overexpression of HSP27 may serve as a biomarker for early detection and therapeutic targets unique to the metastatic phenotype of HCC. PMID- 16261349 TI - A fine physical map of the rice chromosome 5. AB - A fine physical map of the rice (Oryza sativa spp. Japonica var. Nipponbare) chromosome 5 with bacterial artificial chromosome (BAC) and PI-derived artificial chromosome (PAC) clones was constructed through integration of 280 sequenced BAC/PAC clones and 232 sequence tagged site/expressed sequence tag markers with the use of fingerprinted contig data of the Nipponbare genome. This map consists of five contigs covering 99% of the estimated chromosome size (30.08 Mb). The four physical gaps were estimated at 30 and 20 kb for gaps 1-3 and gap 4, respectively. We have submitted 42.2-Mb sequences with 29.8 Mb of nonoverlapping sequences to public databases. BAC clones corresponding to telomere and centromere regions were confirmed by BAC-fluorescence in situ hybridization (FISH) on a pachytene chromosome. The genetically centromeric region at 54.6 cM was covered by a minimum tiling path spanning 2.1 Mb with no physical gaps. The precise position of the centromere was revealed by using three overlapping BAC/PACs for approximately 150 kb. In addition, FISH results revealed uneven chromatin condensation around the centromeric region at the pachytene stage. This map is of use for positional cloning and further characterization of the rice functional genomics. PMID- 16261351 TI - Endoscopic peroneal retinaculum reconstruction. AB - This is a technical note on endoscopic peroneal retinaculum reconstruction in the management of peroneal tendons instability. The surgical trauma of open peroneal retinaculum reconstruction can be minimized with endoscopic approach. The author describes the surgical principle and technique of this endoscopic approach. PMID- 16261350 TI - Bilateral painful snapping elbows triggered by daily dumbbell exercises: a case report. AB - We treated a 64-year-old female with bilateral painful snapping elbows due to synovial folds. Resection of the bilateral synovial folds resulted in relief from pain and snapping, and resulted in patient satisfaction. Daily dumbbell exercises of the bilateral elbows starting at an elderly age may have led to the bilateral snapping. Painful snapping elbow is a relatively rare condition, which usually occurs in a unilateral elbow of a younger patient. To our knowledge, bilateral painful snapping elbows triggered by daily dumbbell exercises in older patients have not been previously reported. PMID- 16261352 TI - Screw fixation of a 4 1/2-year-old PCL avulsion injury. AB - We report about the case of a 4(1/2)-year delayed screw fixation of a tibial posterior cruciate ligament (PCL) avulsion fracture in combination with a posterolateral stabilization. After open reduction and rigid screw fixation, combined with a posterolateral stabilization using a semitendinosus tendon autograft, the patient returned to full activity in sports and daily life. If the PCL substance is sufficient, a delayed refixation in an old PCL avulsion injury seems to be a viable alternative to PCL reconstruction. PMID- 16261353 TI - Intramuscular immunization with p36(LACK) DNA vaccine induces IFN-gamma production but does not protect BALB/c mice against Leishmania chagasi intravenous challenge. AB - Acute visceral leishmaniasis is a progressive disease caused by Leishmania chagasi in South America. The acquisition of immunity following infection suggests that vaccination is a feasible approach to protect against this disease. Since Leishmania homologue of receptors for activated C kinase (LACK) antigen is of particular interest as a vaccine candidate because of the prominent role it plays in the pathogenesis of experimental Leishmania major infection, we evaluated the potential of a p36(LACK) DNA vaccine in protecting BALB/c mice challenged with L. chagasi. In this study, mice received intramuscular (i.m.) or subcutaneous (s.c.) doses of LACK DNA vaccine. We evaluated the production of vaccine-induced cytokines and whether this immunization was able to reduce parasite load in liver and spleen. We detected a significant production of interferon gamma by splenocytes from i.m. vaccinated mice in response to L. chagasi antigen and to rLACK protein. However, we did not observe a reduction in parasite load neither in liver nor in the spleen of vaccinated animals. The lack of protection observed may be explained by a significant production of IL-10 induced by the vaccine. PMID- 16261354 TI - Humoral and cytokine response during protection of mice against secondary hydatidosis caused by Echinococcus granulosus. AB - Infection of BALB/c mouse with protoscoleces of Echinococcus granulosus constitutes a model for the study of secondary hydatidosis and the associated immune response in immunization and infection trials. The aims of this study were to induce a protective immunity against secondary hydatidosis using conventional vaccination approaches and to analyse the immune responses that accompany this protection. Mice immunized with antigen B (AgB), a component of crude sheep hydatid fluid (CSHF), showed a significant level of protection as indicated by a 98.3% reduction in cyst load. This reduction in cyst development was accompanied by a high concentration of interferon gamma secreted by antigen-stimulated spleen cells, as compared with those secreted by cells of mice immunized with CSHF or protoscoleces homogenate (PSH) antigens. In contrast, interleukin-4 was significantly higher in the supernatants of cells stimulated with CSHF or PSH compared with AgB (191.5, 195.7 and 127.5 pg, respectively). Kinetic analysis of immunoglobulin subclasses showed persistently high levels of IgG1 and IgG2a subclasses in immunized infected animals until 6 months of infection, whereas IgG3 showed a significant decline after 1 month of infection. In infected non immunized control mice, all IgG subclasses showed a gradual increase after the first month of infection until the experiment termination (8 months after infection). PMID- 16261355 TI - ITS1 sequence variabilities correlate with 18S rDNA sequence types in the genus Acanthamoeba (Protozoa: Amoebozoa). AB - The subgenus classification of the ubiquitously spread and potentially pathogenic acanthamoebae still poses a great challenge. Fifteen 18S rDNA sequence types (T1 T15) have been established, but the vast majority of isolates fall into sequence type T4, and so far, there is no means to reliably differentiate within T4. In this study, the first internal transcribed spacer (ITS1), a more variable region than the 18S rRNA gene, was sequenced, and the sequences of 15 different Acanthamoeba isolates were compared to reveal if ITS1 sequence variability correlates with 18S rDNA sequence typing and if the ITS1 sequencing allows a differentiation within T4. It was shown that the variability in ITS1 is tenfold higher than in the 18S rDNA, and that ITS1 clusters correlate with the 18S rDNA clusters and thus corroborate the Acanthamoeba sequence type system. Moreover, high sequence dissimilarities and distinctive microsatellite patterns could enable a more detailed differentiation within T4. PMID- 16261357 TI - A new species of the medicinal leech (Oligochaeta, Hirudinida, Hirudo) from Transcaucasia and an identification key for the genus Hirudo. AB - A recent molecular phylogenetic study has suggested that the genus Hirudo contains a neglected species previously known as the orientalis coloration type of the medicinal leech Hirudo medicinalis. In this paper, the new species is formally described as Hirudo orientalis sp. n. It can most readily be identified by the grass green coloration of the dorsum, segmentally arranged pairs of black quadrangular or rounded dots on its paramarginal dorsal stripes and similarly arranged, but less regular light-colored markings on the predominantly black venter. It has medium-sized epididymes and an evenly coiled vagina. H. orientalis is known from Transcaucasia, Iran, and Uzbekistan. It is widely used in medicine as the "medicinal leech." Very little is known about its exact distribution, specific habitat, and conservation status. The paper contains an identification key to all species of the genus Hirudo. PMID- 16261356 TI - Identification and determination of the viability of Giardia lamblia cysts and Cryptosporidium parvum and Cryptosporidium hominis oocysts in human fecal and water supply samples by fluorescent in situ hybridization (FISH) and monoclonal antibodies. AB - In the present study, fluorescent in situ hybridization (FISH) and monoclonal antibodies (MAbs) were evaluated for species-specific detection and viability determination of Giardia lamblia, Cryptosporidium parvum, and Cryptosporidium hominis in human fecal and water supply samples. A total of 50 fecal human samples positive for G. lamblia cysts, 38 positive for C. parvum, and 23 positive for C. hominis were studied. Also, 18 water supply samples positive for Giardia spp. and Cryptosporidium spp. by the United States Environmental Protection Agency (USEPA) Method 1623 were studied by FISH and fluorescein isothiocyanate (FITC)-conjugated MAbs. Eighteen percent of the fecal samples parasitologically positive for G. lamblia presented viable and nonviable cysts, and 5% of those positive for Cryptosporidium spp. presented viable and nonviable oocysts. Of the 18 water supply samples analyzed, 6 (33%) presented Giardia spp. viable and nonviable cysts and 2 (11%) presented viable and nonviable Cryptosporidium spp. oocysts. G. lamblia identification was confirmed by polymerase chain reaction (PCR) and sequencing of the beta-giardin gene in the fecal and water samples found positive by FISH and FITC-conjugated MAbs. C. parvum and Cryptosporidium muris were identified, by PCR and sequencing of the small subunit of ribosomal RNA gene, in seven and one water samples, respectively. Our results confirm that this technique enables simultaneous visualization, species-specific identification, and viability determination of the organisms present in human fecal and water supply samples. PMID- 16261358 TI - Desulfovibrionales-related bacteria in a paper mill environment as detected with molecular techniques and culture. AB - The aim of the present study was to evaluate the suitability of a nested PCR-DGGE (denaturing gradient gel electrophoresis) method for the detection of Desulfovibrionales-related sulfate-reducing bacteria (SRB) from paper mill samples. The samples were also analyzed with culturing. SRB cause/enhance industrial problems, namely creation of foul-smelling gases (hydrogen sulfide) and biological corrosion, and so far there has not been a simple method to study these bacteria in paper mill laboratories. In our study, culturing was able to detect Desulfovibrionales-related bacteria from two different white waters, two different brokes, pulp, clay, and slime. Out of the isolated Desulfovibrionales, 23 enrichment cultures were further characterized with Desulfovibrionales selective PCR-DGGE. An identical Desulfovibrio species sequence was found from paper machine I (broke I, slime, and pulp) and from paper machine II (broke II and white water II), suggesting an in-house contamination with the same strain. Desulfovibrionales-selective PCR-DGGE was also performed from DNA templates extracted directly from the paper mill samples. The DGGE profiles derived from the samples without prior enrichment were more diverse and the sequenced amplicons proved to belong to the Desulfovibrionales order. Moreover, molecular techniques were able to detect Desulfovibrionales-related bacteria from calcium carbonate samples whereas culture did not. Altogether, the nested PCR-DGGE method used in this study was suitable for the detection of Desulfovibrionales-related SRB directly from different paper mill samples and it could be used for the rapid identification of SRB-contaminated industrial sites and, when combined with sequencing, for tracing of the contamination routes. PMID- 16261359 TI - Heterologous production of daptomycin in Streptomyces lividans. AB - Daptomycin and the A21978C antibiotic complex are lipopeptides produced by Streptomyces roseosporus and also in recombinant Streptomyces lividans TK23 and TK64 strains, when a 128 kbp region of cloned S. roseosporus DNA containing the daptomycin gene cluster is inserted site-specifically in the phiC31 attB site. A21978C fermentation yields were initially much lower in S. lividans than in S. roseosporus, and detection was complicated by the production of host metabolites. However A21978C production in S. lividans was improved by deletion of genes encoding the production of actinorhodin and by medium optimization to control the chemical form of the calcium dependent antibiotic (CDA). This latter compound has not previously been chemically characterized as a S. lividans product. Adding phosphate to a defined fermentation medium resulted in formation of only the phosphorylated forms of CDA, which were well separated from A21978C on chromatographic analysis. Adjusting the level of phosphate in the medium led to an improvement in A21978C yield from 20 to 55 mg/l. PMID- 16261361 TI - Artemisinin and thiabendazole are potent inhibitors of cytochrome P450 1A2 (CYP1A2) activity in humans. AB - OBJECTIVE: To investigate the likelihood of artemisinin and thiabendazole causing pharmacokinetic interactions involving cytochrome P450 (CYP1A2) in humans given their potent inhibitory effects on the isoform in vitro. METHODS: Ten healthy volunteers received caffeine (136.5 mg), and after a washout period of 48 h, the volunteers were given a caffeine tablet (136.5 mg) together with thiabendazole (500 mg). After an additional 14 days, the volunteers received caffeine together with artemisinin (500 mg). After each treatment, plasma was obtained up to 24 h post-dose. The plasma concentrations of the drugs were measured by HPLC with UV and MS detection. RESULTS: Using the ratio of paraxanthine to caffeine after 4 h as an indicator of CYP1A2 activity, thiabendazole and artemisinin inhibited 92 and 66%, respectively, of the enzyme activity in vivo. In addition, the pharmacokinetics of caffeine were altered in the presence of the drugs; increases in AUC(0-24) of 1.6-fold (P < 0.01) and 1.3-fold of caffeine in the presence of thiabendazole and artemisinin respectively were measured. The use of in vitro data to predict the effects of thiabendazole on the formation of paraxanthine yielded good results and underestimated the effects of artemisinin when total plasma concentrations were used. Corrections for protein binding resulted in underestimation of inhibitory effects on CYP1A2. CONCLUSIONS: Co-administration of thiabendazole or artemisinin with CYP1A2 substrates could result in clinically significant effects. Our results highlight the validity of in vitro data in predicting in vivo CYP inhibition. The formation of paraxanthine seems to be a better indicator of in vivo CYP1A2 activity than caffeine levels. PMID- 16261362 TI - Changes to methadone clearance during pregnancy. AB - OBJECTIVE: Measurement of plasma methadone concentration to investigate the rate of clearance of methadone prescribed for heroin dependence in the first, second and third trimesters of pregnancy. A secondary objective was to evaluate the outcome of pregnancy. METHODS: Longitudinal within subject study of nine pregnant opioid dependent subjects prescribed methadone at the Leeds Addiction Unit, an outpatient community based treatment centre. Plasma concentration versus time data for methadone was collected during each trimester and post-partum for our subjects. Data was available for the first and second trimesters for 4/9 cases. All but one of the subjects provided data during the third trimester and data post-partum was collected from three respondents. Measurements of methadone levels in plasma were carried out using high performance liquid chromatography (HPLC). RESULTS: Trough mean plasma methadone concentrations reduced as the pregnancies progressed from 0.12 mg/L (first trimester) to 0.07 mg/L (third trimester). The weight-adjusted clearance rates gradually increased from a mean of 0.17 to 0.21 L/hr/kg during pregnancy, although patterns differed substantially between the nine women. An assessment of relative clearance of methadone using two patients for whom we have had all three CL values (trimester 1-3) demonstrated notable change of CL (P = 0.056) over time. Eight of our subjects delivered (3 males), within two weeks of their due date the ninth (male) was premature (21 days). The mean length of gestation was 39.7 weeks (SD = 10 days) and none of the neonates met criterion for 'low birth weight' mean = 3094, SD = 368 g). Five neonates spent time (0.5-28 days) in a special care baby unit (SCUBU) and 4 of these displayed signs of methadone withdrawal. CONCLUSIONS: General Practitioners and hospital doctors should recognise the significant benefits of prescribing methadone for heroin-dependent women during pregnancy. We recommend that if a pregnant opioid user complains of methadone withdrawal symptoms (i.e. that the methadone dose does not "hold" them) the prescribing clinician takes this observation seriously and considers a more detailed assessment. Further work on key factors undergoing changes during pregnancy accounting for differences in methadone metabolism in the mother, fetus and neonate are required. PMID- 16261363 TI - Pharmacokinetics and pharmacodynamics of etizolam are influenced by polymorphic CYP2C19 activity. AB - OBJECTIVE: To examine the effect of cytochrome P450 (CYP) 2C19 activity on the single-dose pharmacokinetics and pharmacodynamics of etizolam. METHODS: The subjects were 21 healthy Japanese volunteers. The two mutated alleles (CYP2C19*2 and CYP2C19*3) causing absent CYP2C19 activity were identified by a polymerase chain reaction method. Twelve subjects were extensive metabolizers (EMs) with no or one mutated allele, and nine subjects were poor metabolizers (PMs) with two mutated alleles. The subjects received a single oral 1-mg dose of etizolam, and blood samplings and evaluation of psychomotor function were conducted up to 24 h after dosing. RESULTS: The PMs had significantly larger total area under the plasma concentration-time curve (287+/-74 vs 178+/-122 ng.h/ml, p<0.05) and longer elimination half-life (14.8+/-4.2 vs 10.5+/-3.9 h, p<0.05) of etizolam than the EMs. The area under the score-time curve from 0 to 8 h of the Stanford Sleepiness Scale was significantly larger in the PMs than in EMs (28.9+/-5.2 vs 22.9+/-6.9 score.h, p<0.05). CONCLUSION: The present study suggests that the single-dose pharmacokinetics and pharmacodynamics of etizolam are influenced by polymorphic CYP2C19 activity. PMID- 16261364 TI - Phase I dose-finding study and a pharmacokinetic/pharmacodynamic analysis of the neutropenic response of intravenous diflomotecan in patients with advanced malignant tumours. AB - PURPOSE: To determine the maximum tolerated dose (MTD) of intravenous (iv) diflomotecan administered once every 3 weeks, and to characterize the relationship between pharmacokinetics and neutropenic effect, using a semi mechanistic pharmacokinetic/pharmacodynamic (PK/PD) model. EXPERIMENTAL DESIGN: Twenty-four patients received a total of 75 cycles of iv diflomotecan that was administered as 20-min infusion, once every 3 weeks at escalating doses of 2, 4, 5, and 6 mg/m2. Haematological and non-haematological toxicities were evaluated. Plasma concentrations of diflomotecan were measured after the first drug administration. RESULTS: Dose limiting toxicity (DLT) following the first cycle occurred in 12 patients and a total of 16 patients experienced DLT at some point in the trial. During the first cycle of treatment the number of patients in the 5 and 6 mg/m2 dose groups that experienced DLT was 3 of 4, and 3 of 3, respectively. Therefore, the dose of 5 mg/m2 was considered the MTD and the dose of 4 mg/m2 the recommended dose (RD). During the first cycle, 12 patients experienced DLT, six had either infection of haematological toxicity and eight complained of fatigue. The best response was a partial response in one patient treated at the 6 mg/m2 dose level. Disease stabilization was observed in seven patients (four patients treated at 4 mg/m2 and one patient at each dose level of 2, 5, and 6 mg/m2). The remaining patients had all progressive disease. The median time to progression for all patients was 5.9 weeks. Pharmacokinetics of diflomotecan was described with a three-compartmental model. Mean population parameter estimates of the apparent volume of distribution of the central compartment (V c) increased linearly with body surface area (BSA) as: V c (L) = 41.5 x (BSA/1.85), and the mean population estimate of the apparent volume of distribution of the shallow compartment was lower in females (29.5 vs 48.8 L). Computer simulations showed the lack of clinical significance of these covariates. The time course of the neutropenic response was adequately described by a semi-mechanistic model that includes cellular processes and drug effects. CONCLUSIONS: The MTD and RD after a 20-min iv infusion of diflomotecan every 3 weeks are 4 and 5 mg/m2, respectively. Diflomotecan showed linear pharmacokinetic behaviour and the selected PK/PD model described adequately the time course of neutropenia. The mean model predicted values of nadir and time to nadir after a 20-min iv infusion of 4 mg/m2 of diflomotecan was 0.86 x 10(9) /L neutrophil cell counts and 11 days, respectively. PMID- 16261366 TI - Do early injections of bulking agents following radical prostatectomy improve early continence? AB - Bulking agents belong to the treatment options of stress urinary incontinence in both genders for a long time, but early injection of bulking agents following radical prostatectomy to improve early continence is a new treatment modality and the results of the first years will be presented in this study. From 1996 to 2001, 55 incontinent patients, of whom 44 suffered from post-prostatectomy incontinence (PPI), have been treated by bulking agents in our clinic and evaluated later on, using a semistandardized questionnaire. Mean age was 65.5 years and the mean follow-up was 32 months. Of 44 patients with PPI, 29 suffered from incontinence of the third, 10 from incontinence of the second and 5 from incontinence of the first degree. We examined the influence of transurethral injection of bulking agents on direct postoperative continence (short-term results) and on continence at the time of follow-up (long-term results) (mean 32 month). An improved early continence (short-term result) was achieved in 30 patients. At follow-up, 19 patients were completely dry, 18 reported an improved continence. Especially the time of treatment, referring to radical prostatectomy, had a significant influence on treatment results with better results in the subgroup of 34 patients that have been treated early (mean 23 days) compared to 10 patients that received delayed treatment (mean 26 months) following radical prostatectomy. The preoperative degree of incontinence had a significant influence on short-term results, and not on long-term results. The amount of bulking material and the number of injections had no significant influence on short-term results, but repeated injections resulted in statistically worse long term results. In summary, early transurethral injection of bulking agents is an effective and minimal invasive therapy option to improve early continence in patients with PPI. Time to improved continence due to training of the sphincter externus muscle cannot be shortened, but the quality of life during this time can be improved. In case of a severe lesion of the sphincteric muscle, continence cannot be improved for a longer period and the results are poor. In these cases, procedures like implantation of an artificial urinary sphincter may be required. PMID- 16261365 TI - Local anesthesia for transurethral manipulations: is a transrectal periprostatic nerve block effective? AB - Periprostatic nerve block has been reported to be an effective form of anesthesia for prostatic surgery. Recent studies have shown the simplicity and the efficacy of a transrectal approach for periprostatic nerve block. The goal of our study was to evaluate the effect of a transrectal periprostatic nerve block on the discomfort associated with rigid cystoscopy. A total of 73 patients underwent cystoscopy. Group 1 (n = 39) received a transrectal periprostatic lidocaine infiltration prior to the cystoscopy. Group 2 (n = 34) underwent cystoscopy alone. The pain that patients experienced during cystoscopy was assessed on a visual analog scale. The patients in the two groups were very similar in regard to age and size of the prostate. The mean pain score was 3.4 in group 1 and 3.9 in group 2. This difference was not statistically significant. We conclude that nerve block does not seem to reduce the pain associated with transurethral manipulations. PMID- 16261367 TI - Production and characterization of laccase from Cyathus bulleri and its use in decolourization of recalcitrant textile dyes. AB - Many fungi (particularly the white rot) are well suited for treatment of a broad range of textile dye effluents due to the versatility of the lignin-degrading enzymes produced by them. We have investigated decolourization of a number of recalcitrant reactive azo and acid dyes using the culture filtrate and purified laccase from the fungus Cyathus bulleri. For this, the enzyme was purified from the culture filtrate to a high specific activity of 4,022 IU mg(-1) protein, produced under optimized carbon, nitrogen and C/N ratio with induction by 2,6 dimethylaniline. The protein was characterized as a monomer of 58+/-5.0 kDa with carbohydrate content of 16% and was found to contain all three Cu(II) centres. The three internal peptide sequences showed sequence identity (80-92%) with laccases of a number of white rot fungi. Substrate specificity indicated highest catalytic efficiency (k(cat)/K(M)) on guaiacol followed by 2,2'-azino-bis(3 ethylthiazoline-6-sulfonic acid) (ABTS). Decolourization of a number of reactive azo and acid dyes was seen with the culture filtrate of the fungus containing predominantly laccase. In spite of no observable effect of purified laccase on other dyes, the ability to decolourize these was achieved in the presence of the redox mediator ABTS, with 50% decolourization in 0.5-5.4 days. PMID- 16261368 TI - MCNP5 evaluation of dose dissipation in tissue-like media exposed to low-energy monoenergetic X-ray microbeam. AB - Following a significant increase in the number of facilities in the world having and developing low- and high-linear energy transfer (LET) microbeams for experimental radiobiological studies, it is useful and demanding to establish reliable computational models to analyze such experiments. This paper summarizes initial MCNP5 calculations of the basic parameters needed to study X-ray microbeam penetration, dose deposition and dose spatial dissipation in tissue like media of micro and macro scales. The presented models can be used to predict doses delivered to neighboring cells and analyze the cause of bystander cell deaths. In the case of low-LET radiation, dose distribution is more homogenized when compared to high-LET that deposits almost all of its energy in the cell hit by radiation. Results are presented for a microbeam of monoenergetic soft (2-10 keV) X-rays for two different micro-models: (a) single-cells of homogeneous and uniform chemical compositions, and (b) single-cells of heterogeneous structures (nucleus and cytoplasm) with different chemical compositions. In both numerical models, only one cell is irradiated and the electron and X-ray doses in all cells are recorded. It was found that surrounding cells receive approximately five orders of magnitude less dose than the target cell in the homogenized cell model. The more detailed, heterogeneous model showed that the nucleus of the target cell receives more than 95% of the dose delivered to the entire cell, while neighboring cell nuclei receive approximately 65% of their total cell dose. Results of the macroscopic behavior of a soft X-ray microbeam using a cylindrical phantom 5 cm tall and 1 cm in diameter are also presented. Three-dimensional dose profiles indicate the spatial dose dissipation. For example, a 10 keV X-ray microbeam dose scatters to a negligible level at 0.3 cm radially from the center while it reaches an axial depth of 2 cm. PMID- 16261369 TI - Orally active antioxidative copper(II) aspirinate: synthesis, structure characterization, superoxide scavenging activity, and in vitro and in vivo antioxidative evaluations. AB - Ever since it was proposed that reactive oxygen species (ROS) are involved in the pathogeneses of various diseases, superoxide dismutase (SOD)-mimetic complexes have been intensively studied. We prepared copper(II) aspirinate [Cu2(asp)4] from Cu(II) and aspirin, which has been in use for many years as an antipyretic, an analgesic, and an anti-inflammatory agent. However, Cu2(asp)4 has been found to have additional activities, including anti-inflammatory, antiulcer, anti ischemic/reperfusion agent, anticancer, antimutagenic, and antimicrobial activities. The activity of copper salicylate [Cu(sal)2] was also compared with that of Cu2(asp)4. The structure of the Cu2(asp)4 was determined using X-ray structure analysis. Its SOD-mimetic activity was determined using cytochrome c, electron spin resonance (ESR) spectroscopy, and ESR spin trap methods. The activity of Cu2(asp)4 was slightly greater than CuSO4 and copper acetate [Cu(ace)2] and slightly less than that of Cu(sal)2. The in vitro antioxidant activity, evaluated in human epithelial or transformed neoplastic keratinocyte cells, HaCaT, and normal dermal fibroblasts in terms of cell survival following ultraviolet B (UVB) irradiation, was significantly increased in the presence of Cu2(asp)4, Cu(sal)2, and CuSO4. Further, ROS generation following UVA irradiation in the skin of hairless mice following oral treatment with Cu2(asp)4 for three consecutive days was significantly suppressed compared to the vehicle- or Cu(ace)2-treated mice. On the basis of these results, Cu2(asp)4 was observed to be a potent antioxidative compound possessing antioxidative activity in biological systems. In conclusion, Cu2(asp)4 is a potent antioxidative agent that may be useful for future treatment of diseases resulting from ROS. PMID- 16261370 TI - Thyroglossal duct cyst's inflammation. When do we operate? AB - Thyroglossal duct cyst (TGDC) disease, one of the most common developmental neck lesion in the pediatric population, often presents as infected neck mass. The authors reviewed their experience in the management of inflamed TGDC cases, in order to suggest the most efficient approach of these patients regarding the ideal type and time of surgical intervention. The medical records of all the patients with the diagnosis of TGDC treated at our department from 1988 to 2003 were reviewed. Data collected included age, gender, preoperative inflammation, treatment and time of definitive surgery. The outcome of the operation was graded as successful, recurrence, or postoperative infection. Eighty-nine (89) patients with histologically confirmed TGDC were treated at our department. Mean age at operation was 6.0 years (range, 9 months-14 years). Male to female ratio was 1.2:1. All patients underwent Sistrunk operation and the mean follow-up was 3 years. Fifty-four (54) patients (60.6%) presented with an inflamed TGDC. Among them, 24 patients were operated immediately after diagnosis at the phase of acute inflammation, and 30 patients after antibiotic administration and resolution of inflammation. The overall recurrence incidence was 6.7%. Recurrence was noted in 6 of 24 (25%) patients operated during the acute phase of inflammation and none in patients operated after resolution of inflammation (P = 0.0052) or with no preoperative inflammation (P = 0.0002). Postoperative wound infection was noted in seven cases but none of them developed recurrence of the disease. The presence of inflammation at the time of surgery is an important risk factor for relapse. We suggest that in the inflamed cases of TGDC disease, the initial treatment should be antibiotic administration and after resolution of the inflammation, surgical management should follow. PMID- 16261371 TI - Down's syndrome and esophageal achalasia: a rare but important clinical entity. AB - We report a case of esophageal achalasia (EA) in Downa9s syndrome. A six-year-old girl with DS was referred from another hospital for further management of dysphagia and growth retardation (weight 16.3 kg, height 105.4 cm: both below 1.5 SD). Dysphagia commenced when she was one year old and gastroesophageal reflux (GER) was diagnosed when she was four. Routine investigations suggested EA or esophageal stenosis secondary to GER. While dissecting the esophago-gastric junction in preparation for a Heller-Dor esophagocardiomyotomy (HD-ECM), the crus of the diaphragm was noted to be narrowed, severely fibrosed and attached to the lower esophagus which was covered by dense scar tissue suggestive of an old esophageal perforation secondary to GER esophagitis. A Nissen fundoplication was performed, but dysphagia persisted postoperatively, and a narrowing 2 cm above the fundoplication wrap with proximal dilatation was found on repeated barium studies. At re-laparotomy, the fundoplication was unwrapped and HD-ECM performed. Eight months postoperatively, she is well with no recurrence. EA must be considered in any DS patient presenting with dysphagia. PMID- 16261372 TI - Effect of TNF-alpha on Raji cells at different cellular levels estimated by various methods. AB - Tumor necrosis factor (TNF)-alpha, a pleiotropic cytokine, has been shown to induce diverse and opposite effects on lymphoid malignancy depending on TNF receptor system expression. Based on this, we investigated its in vitro dose- and time-related effect on the malignant B-cell line Raji, derived from Burkitt lymphoma patients, at different intracellular levels. The membrane alteration was estimated by lactate dehydrogenase (LDH) release and by flow cytometry; intracellular metabolic energy by determination of the total intracellular LDH activity; total cytosole protein mass by sulforhodamine B assay; and cell growth by incorporation of [3H]thymidine into DNA. Significant increase of LDH through cell membrane alteration was accompanied by decrease of intracellular metabolized energy and total protein mass. TNF-alpha at lower concentrations (125 and 250 pg/ml) significantly induced cell proliferation in comparison with 1,000 pg/ml of TNF-alpha, which induced more cell death. TNF-alpha induced maximal apoptosis rate up to 30% after 24 h, showing more effects for a necrotic form of cell death. Here we reported opposite and diverse effects of TNF-alpha at different intracellular levels in Raji cells, when applied in different assays, showing characteristics for every cellular compartment. PMID- 16261373 TI - Effective strategies for forensic analysis in the mitochondrial DNA coding region. AB - Recently, it has been recognized that accessing information in the mtDNA coding region can provide additional forensic discrimination with respect to the standard typing of the D-loop region, augmenting the sometimes rather limited forensic power of mtDNA testing. Here, we discuss considerations relating to maximally effective approaches for recovering additional discrimination in the coding region, bearing in mind that (1) DNA quality and quantity in typical mtDNA casework usually restrict the amount of additional sequence that can be obtained, and (2) the need for additional discrimination primarily arises when common HV1/HV2 types are encountered. Most investigators have sought additional discrimination by sequencing short segments of coding region that are thought to be particularly variable. Unfortunately, efforts in this regard have generally failed to appreciate that most variation in the coding region is redundant with information already present in HV1/HV2 and have therefore overvalued the potential of this approach for providing additional discrimination. An alternative single nucleotide polymorphism-based approach [Int J Legal Med 118:137-146, 2004] has been to identify specific bases that provide resolution in specific common HV1/HV2 types (and related sequences). We investigate several highly relevant data sets wherein the latter approach performs appreciably better than sequencing selected short portions of the coding region. This is true even when only synonymous variation is targeted to minimize the potential for problems arising from discovery of mutations that have reportedly been related to disease. PMID- 16261374 TI - Filaggrin repeat number polymorphism is associated with a dry skin phenotype. AB - Profilaggrin is a key epidermal protein, critical for the generation and maintenance of the stratum corneum barrier. It is encoded by a gene located in the epidermal differentiation complex of Chromosome 1q21 and is composed of multiple filaggrin repeats connected by highly conserved linker peptides. Within the human population the number of filaggrin repeats encoded by this gene varies between 10, 11 or 12 repeats. Using a PCR-based approach we have determined individual profilaggrin allelotypes in a group of 113 subjects and identified preliminary evidence of an inverse association between the 12 repeat allele and self-perceived frequent dry skin (P=0.0293). This is the first demonstration of a potential association between a genetic marker and cosmetic skin condition and suggests that cosmetic skin dryness may in part be genetically determined and associated with specific profilaggrin allelotypes. PMID- 16261375 TI - Sucrose synthase and RuBisCo expression is similarly regulated by the nitrogen source in the nitrogen-fixing cyanobacterium Anabaena sp. AB - In higher plants and cyanobacteria, sucrose (Suc) metabolism is carried out by a similar set of enzymes. The function and regulation of Suc metabolism in cyanobacteria has begun to be elucidated. In strains of Anabaena sp., filamentous nitrogen-fixing cyanobacteria, Suc synthase (SuS, EC 2.4.1.13) controls Suc cell level through the cleavage of the disaccharide. The present work shows that there are two sus genes in Anabaena (Nostoc) sp. that are co-regulated regarding the nitrogen source; however, only susA accounts for the extractable SuS activity and for the control of the Suc level. Primer extension analysis has uncovered the sequence of the Anabaena susA and susB ammonium-activated putative promoters, which share a high sequence similarity with that of rbcLS encoding ribulose bisphosphate carboxylase/oxygenase (EC 4.1.1.39) and other ammonium up-regulated genes. Moreover, susA and rbcLS expression is developmentally co-localized to the vegetative cells of the nitrogen-fixing cyanobacterial filaments. Our results strongly suggest the existence of a regulatory network that would coordinate the expression of key genes for Suc and nitrogen metabolism, carbon fixation, and development in Anabaena sp. PMID- 16261376 TI - Large herbivores influence the composition and diversity of shrub-steppe communities in the Rocky Mountains, USA. AB - It is widely believed that wild and domestic herbivores have modified the structure and composition of arid and semi-arid plant communities of western North America, but these beliefs have rarely been tested in long-term, well replicated studies. We examined the effects of removing large herbivores from semi-arid shrublands for 40-50 years using 17 fenced exclosures in western Colorado, USA. Shrub cover was greater (F=5.87, P=0.0020) and cover (F=3.01, P=0.0601) and frequency (F=3.89, P=0.0211) of forbs was less inside the exclosures (protected) relative to grazed plots. However, we found no significant effects (minimum P=0.18) of protection from grazing on cover or frequency of grasses, biotic crusts, or bare soil. Although mean species richness and diversity were similar between treatments, protected areas had much higher dominance by fewer species, primarily sagebrush. Exclusion of herbivores changed the relationship between species richness and evenness. Consistent with theoretical expectations, species evenness was positively correlated with richness in protected plots (r2=0.54). However, contrary to theory, evenness and richness were inversely related in grazed plots (r2adjacent=0.72, r2distant=0.84). We suggest that these differences resulted because grazing acts as a stressor promoting facilitative relationships between plant species that might compete for resources in the absence of grazing. We conclude that exclusion of grazing in the sites we studied caused minor changes in cover and diversity of herbaceous plants, but caused a clear increase in the cover of shrubs. Importantly, the exclusion of ungulates changed the relationship between evenness and richness. PMID- 16261377 TI - Fitness consequences for copepods feeding on a red tide dinoflagellate: deciphering the effects of nutritional value, toxicity, and feeding behavior. AB - Phytoplankton exhibit a diversity of morphologies, nutritional values, and potential chemical defenses that could affect the feeding and fitness of zooplankton consumers. However, how phytoplankton traits shape plant-herbivore interactions in the marine plankton is not as well understood as for terrestrial or marine macrophytes and their grazers. The occurrence of blooms of marine dinoflagellates such as Karenia brevis suggests that, for uncertain reasons, grazers are unable to capitalize on, or control, this phytoplankton growth-making these systems appealing for testing mechanisms of grazing deterrence. Using the sympatric copepod Acartia tonsa, we conducted a mixed diet feeding experiment to test whether K. brevis is beneficial, toxic, nutritionally inadequate, or behaviorally rejected as food relative to the palatable and nutritionally adequate phytoplankter Rhodomonas lens. On diets rich in K. brevis, copepods experienced decreased survivorship and decreased egg production per female, but the percentage of eggs that hatched was unaffected. Although copepods showed a 6 17% preference for R. lens over K. brevis on some mixed diets, overall high ingestion rates eliminated the possibility that reduced copepod fitness was caused by copepods avoiding K. brevis, leaving nutritional inadequacy and toxicity as remaining hypotheses. Because egg production was dependent on the amount of R. lens consumed regardless of the amount of K. brevis eaten, there was no evidence that fitness costs were caused by K. brevis toxicity. Copepods limited to K. brevis ate 480% as much as those fed only R. lens, suggesting that copepods attempted to compensate for low food quality with increased quantity ingested. Our results indicate that K. brevis is a poor food for A. tonsa, probably due to nutritional inadequacy rather than toxicity, which could affect bloom dynamics in the Gulf of Mexico where these species co-occur. PMID- 16261378 TI - Growth, compatible solute and salt accumulation of five mycorrhizal fungal species grown over a range of NaCl concentrations. AB - The oil sand industry in northeastern Alberta produces vast areas of severely disturbed land. The sodicity of these anthropic soils is one of the principal constraints that impede their revegetation. Previous in vitro studies have shown that the ectomycorrhizal fungi Laccaria bicolor (Maire) Orton UAMH 8232 and Hebeloma crustuliniforme (Bull) Quel. UAMH 5247 have certain salt-resistant traits and thus are candidate species for the inoculation of tree seedlings to be outplanted on salt-affected soil. In this study, the in vitro development of these fungi was compared to that of three mycorrhizal fungi [Suillus tomentosus (Kauff.) Sing., Snell and Dick; Hymenoscyphus sp. and Phialocephala sp.] isolated from a sodic site created by Syncrude Canada Ltd. Their growth, osmotica and Na/Cl contents were assessed over a range (0, 50, 100, 200 mM) of NaCl concentrations. After 21 days, the two ascomycetes (Hymenoscyphus sp. and Phialocephala sp.) were shown to be more resistant to the NaCl treatments than the three basidiomycete species. Of the basidiomycetes, L. bicolor was the most sensitive to NaCl stress, while H. crustuliniforme showed greater water stress resistance, and the S. tomentosus isolate exhibited greater Na and Cl filtering capacities and had a better biomass yield over the NaCl gradient tested. Both ascomycetes used mechanisms other than carbohydrate accumulation to palliate NaCl stress. While the Hymenoscyphus isolate accumulated proline in response to NaCl treatments, the darker Phialocephala isolate may have used compounds such as melanin. The basidiomycete species accumulated mainly mannitol and/or proline in response to increasing concentrations of NaCl. PMID- 16261380 TI - Exciting applications of single chain trimers of MHC-I molecules. PMID- 16261379 TI - Intratumour T cells, their activation status and survival in gastric carcinomas characterised for microsatellite instability and Epstein-Barr virus infection. AB - Gastric carcinomas (GCs) with high microsatellite instability (MSI) or an Epstein Barr virus (EBV) infection are prevalently poorly differentiated adenocarcinomas with abundant lymphoid infiltration. The aims of the study were to clarify (1) if tumour-infiltrating lymphocytes (TILs) and cytotoxic-activated TILs are associated with a better clinical outcome in patients with GCs characterised for the presence of MSI and EBV; (2) if the nature and the activation status of TILs are involved in tumour cell apoptosis, evaluated using the M30 antibody, directed against a fragment of cytokeratin-18 caspase-cleaved during early steps of epithelial cell apoptosis. The immunophenotype of TILs and the tumour cell apoptosis were analysed with immunohistochemistry in 96 GCs, including 35 MSI GCs, and 61 GCs without MSI [microsatellite stable (MSS)], 17 of which were EBV+. MSI and MSS/EBV+ GCs displayed a significantly higher mean number of cytotoxic activated TILs and apoptotic tumour cells than MSS/EBV- GCs (CD8+ TILs/HPF, 21.7 and 69.6 vs 6.4; T-cell intracellular antigen (TIA)-1+ TILs/HPF, 16.7 and 32.05 vs 5.2; granzyme B+ TILs/HPF, 7.5 and 8.6 vs 0.8; perforin+ TILs/HPF, 5.9 and 9.2 vs 0.9; and M30 IR tumour cells, 5.9 and 2.9 vs 2.3%). In addition to the most reliable clinico-pathological parameters (lymph node status, depth of tumour invasion and tumour stage), a univariate analysis showed that the presence of CD3+ TILs higher than 14.9 (p=0.01), CD8+ TILs higher than 9.5 (p<0.05) and MSI (p=0.02) were associated with better overall patient survival. Using a Cox regression model, only a high number of CD3+ TILs (p=0.02) and a low tumour stage (p=0.00001) were identified as independent prognostic factors. In conclusion, our study demonstrates that a high number of CD3+ and CD8+ TILs is a characteristic of MSI- and EBV-associated GCs and represents a favourable prognostic factor, independently of the pathogenesis of GCs. PMID- 16261382 TI - Characterization of MHC class II genes from an ancient reptile lineage, Sphenodon (tuatara). AB - The organization and evolution of major histocompatibility complex (MHC) genes vary considerably among vertebrate lineages. MHC genes have been well characterized in mammals, birds, amphibians and fish, but little is known about their organization in reptiles, despite the fact that reptiles occupy an important phylogenetic position for understanding the evolutionary history of both mammalian and avian MHC genes. Here we describe the characterization of the first MHC class II B cDNA sequences from a non-avian reptile, the tuatara (Sphenodon spp.). Three class II B sequences were isolated from a tuatara cDNA library, and four additional partial sequences were isolated by reverse transcriptase-polymerase chain reaction. Six of these sequences appear to belong to the same gene family, which we have named SppuDAB. The remaining sequence (named SppuDBB) shares only 43.9% amino acid similarity with SppuDAB and thus appears to represent a separate gene family. SppuDBB may be a non-classical locus as it does not contain all the conserved residues expected of a classical MHC class II gene. Southern blot analysis indicates that only a single copy of SppuDBB exists in tuatara, but that multiple loci related to SppuDAB are present. The SppuDAB sequences have the highest amino acid similarity (57.2-62.4%) with class II B sequences from the spectacled caiman, but only 26.4-48.7% similarity with sequences from other vertebrates. The tuatara sequences do not strongly group with other reptile sequences on a phylogenetic tree, reflecting the antiquity of the Sphenodon lineage and the lack of closely related sequences for comparison. PMID- 16261381 TI - Computational studies on class I ribonucleotide reductase: understanding the mechanisms of action and inhibition of a cornerstone enzyme for the treatment of cancer. AB - This review provides a synthesis of recent work, using computational methods, on the action and inhibition mechanisms of class I ribonucleotide reductase (RNR). This enzyme catalyzes the rate-limiting step of the pathway for the synthesis of DNA monomers and, therefore, has long been regarded as an important target for therapies aiming to control pathologies that depend strongly on DNA replication. In fact, over the last years, several molecules, which are able to impair RNR activity by different mechanisms, have been applied effectively in anti-cancer, anti-viral and anti-parasite therapies. A better understanding of the chemical mechanisms involved in normal catalysis and in inhibition of the enzyme is important for the rational design of more specific and effective inhibitor compounds. To achieve this goal, computational methods, particularly quantum chemical calculations, have been used more and more frequently. The ever-growing capabilities of these methods together with undeniable advantages make it a stimulating area for research purposes. PMID- 16261383 TI - High level of functional polymorphism indicates a unique role of natural selection at human immune system loci. AB - Several studies have shown that immune system proteins have on average a higher rate of amino acid evolution between different species of mammals than do most other proteins. To test whether immune-system-expressed loci show a correspondingly elevated rate of within-species nonsynonymous (amino acid altering) polymorphism, we examined gene diversity (heterozygosity) at 4,911 single nucleotide polymorphism (SNP) sites at 481 protein-coding loci. At loci with nonimmune functions, gene diversity at nonsynonymous SNP sites was typically lower than that at silent SNP sites (those not altering the amino acid sequence) in the same gene, a pattern that is an evidence of purifying selection acting to eliminate slightly deleterious variants. However, this pattern was not seen at nonsynonymous SNPs causing conservative amino acid replacements in immune system proteins, indicating that the latter are subject to a reduced level of functional constraint. Similarly, immune system genes showed higher gene diversities in their 5' noncoding regions than did other proteins. These results identified certain immune system loci that are likely to be subject to balancing selection that acts to maintain polymorphism in either coding or regulatory regions. PMID- 16261385 TI - Familial Dandy-Walker syndrome: a case report supporting an autosomal inheritance. AB - INTRODUCTION: We report an isolated pedigree in which a consanguineous couple had twin sons with Dandy-Walker malformation (DWM). The mother is similarly affected with the disorder. DISCUSSION: DWM is an abnormality of the central nervous system, which leads to hydrocephalus and is associated with other abnormalities. CONCLUSION: Inheritance of the disorder remains controversial, with the majority perceived to be sporadic cases. This report suggests an autosomal inheritance. PMID- 16261384 TI - Costs in rheumatology: results and lessons learned from the 'Hannover Costing Study'. AB - The objective of this study is to review the concept of the 'Hannover Costing Study' and to present and discuss the major insights generated during the course of the project. The costing study was performed in conjunction with a randomized controlled prospective trial assessing the effectiveness of a disease management module in rheumatoid arthritis (RA). A full set of clinical and cost data both from patient-reported and payer-derived cost data was developed. In particular the study included (1) the development of a matrix of cost domains which might be used as a common taxonomy in costing studies, (2) the descriptive analysis of payer derived cost data, (3) the analysis of cost data in patients with uncertain diagnosis; (4) the development and validation of a patient-reported costing instrument, and (5) an assessment of productivity costs. The following are the results (1) the developed matrix of cost domains included 16 separate cost domains: 7 outpatient, 3 inpatient, 4 other disease related, and 2 productivity domains; (2) the micro-costing analysis showed total direct costs of 3,815 per patient-year (standard error of mean, SEM: 267) and RA-related direct costs were 2,312 per patient-year; (3) in patients with uncertain diagnosis of RA and no treatment with 'Disease Modifying Antirheumatic Drugs' (DMARD) costs were significantly lower; (4) the comparison of patient-reported with payer reported cost data generally supports the use of highly aggregated items to assess health care utilization in RA; (5) productivity costs in patients that are gainfully employed and in patients who receive RA-related retirement payments exceed RA-related direct costs. Furthermore, RA-patients reported their productivity losses adequately. The study added some additional insights to the following questions: What costs should be collected, what level of detail is required for that task, what patients should by analyzed, and what data sources should be used in further studies in RA. PMID- 16261386 TI - Validity of the two-parameter model in estimating the anaerobic work capacity. AB - The curvature of the power-time (P-t) relationship (W') has been suggested to be constant when exercising above critical power (CP) and to represent the anaerobic work capacity (AWC). The aim of this study was to compare W' to (1) the total amount of work performed above CP (W (90s)') and (2) the AWC, both determined from a 90s all-out fixed cadence test. Fourteen participants (age 30.5 +/- 6.5 years; body mass 67.8 +/- 10.3 kg), following an incremental VO(2max) ramp protocol, performed three constant load exhaustion tests set at 103 +/- 3, 97 +/- 3 and 90 +/- 2% P-VO(2max) to calculate W' from the P-t relationship. Two 90s all out efforts were also undertaken to determine W (90s)' (power output-time integral above CP) and AWC (power output-time integral above the power output expected from the measured VO(2)). W' (13.6 +/- 1.3 kJ) and W (90s)' (13.9 +/- 1.1 kJ; P = 0.96) were not significantly different but were lower than AWC (15.9 +/- 1.2 kJ) by 24% (P = 0.03) and 17%, respectively (P = 0.04). All these variables were correlated (P < 0.001) but great extents of disagreement were reported (0.2 +/- 6.4 kJ between W' and W (90s)', 2.3 +/- 7.2 kJ between W' and AWC, and 2.1 +/- 4.3 kJ between W (90s)' and AWC). The underestimation of AWC from both W' and W (90s)' can be explained by the aerobic inertia not taking into consideration when determining the two latter variables. The low extents of agreement between W', W (90s)' and AWC mean the terms should not be used interchangeably. PMID- 16261387 TI - Relationship between erector spinae static endurance and muscle oxygenation-blood volume changes in healthy and low back pain subjects. AB - The purposes of this study were to: (1) compare the muscle blood volume (Mbv) and oxygenation (Mox) responses on the right and left side erector spinae during the Biering-Sorensen muscle endurance (BSME) test between healthy, low back pain active (LBP-A) and LBP-sedentary (LBP-S) subjects using near infrared spectroscopy (NIRS), and (2) determine the relationships between the BSME time and Mbv and Mox. Informed consent was obtained from 30 healthy and 30 chronic LBP subjects. The latter group was subdivided into an active (LBP-A; n = 18) and sedentary (LBP-S; n = 12) subgroups based on physical activity patterns. The groups were age and sex matched. The NIRS probes were placed bilaterally on the erector spinae muscle at the level of the third lumbar vertebrae. The testing protocol was: 2-min resting baseline, BSME to voluntary fatigue, and 4-min recovery. During the BSME Mbv immediately increased for approximately 30-60 s, then showed an asymptote, and a steady decline towards the baseline at termination. Mox demonstrated either an initial increase followed by a systematic decrease until the termination of the BSME, or a systematic decrease throughout the BSME until termination. The LBP-S subjects showed a reduced Mox-range and slower Mox (1/2) recovery time on the left side suggesting a reduced aerobic capacity of the erector spinae muscle in this group. Significant correlations were noted between BSME time and the pooled values of Mox-delta and-range only in the LBP-A group. These observations suggest that factors other than erector spinae aerobic capacity can influence BSME performance. PMID- 16261388 TI - Influence of cyclic mechanical strain and heat of human tendon fibroblasts on HSP 72. AB - Heat shock protein 72 (HSP-72) is a member of a superfamily of different proteins that are synthesized as a cytoprotective response following cellular stress. Mechanical strain is an important component in ligament and tendon healing. Up to the present point of time, the influence of mechanical strain on the expression of HSP-72 is unknown. Tendon fibroblasts from the patellar tendons of nine individuals were isolated and amplified in vitro. First, the effect of 15 or 60 min of heat exposition was studied immunohistochemically and by Western blotting. In a second experiment, the effects of 15 and 60 min of cyclic longitudinal stretching were investigated. Samples were taken after 2, 4 and 8 h. The heat exposition experiments indicate that HSP-72 accumulates in the nucleus and that there is a transient upregulation. This effect is more prominent after 60 min of heat exposure. The same reaction was found after stretching stimulation, however, to a lesser extent. There was a transient up regulation of HSP-72 after short term stretching and a biphasic increase after 60 min of stretching. Upregulation of HSP-72 by heat and mechanical stress is a response in human fibroblasts which involves a nuclear translocation. The response differs with regard to the time points beyond 2 h after the application of either stress. PMID- 16261389 TI - Estimation of maximal oxygen uptake by bioelectrical impedance analysis. AB - Previous non-exercise models for the prediction of maximal oxygen uptake VO(2max) have failed to accurately discriminate cardiorespiratory fitness within large cohorts. The aim of the present study was to evaluate the feasibility of a completely indirect method for predicting VO(2max) that was based on bioelectrical impedance analysis (BIA) in 66 young, healthy fit men and women. Multiple, stepwise regression analysis was used to determine the usefulness of BIA and additional covariates to estimate VO(2max) (ml min(-1)). BIA was highly correlated to VO(2max) (r = 0.914; P < 0.001) and entered the regression equation first. The inclusion of gender and a physical activity rating further improved the model which accounted for 88% of the variance in VO(2max) and resulted in a relative standard error of the estimate (SEE) of 7.2%. Substantial agreement between the methods was confirmed by the fact that nearly all the differences were within +/-2 SD. Furthermore, in contrast to previously published non exercise models, no trend of a reduction in prediction accuracy with increasing VO(2max) values was apparent. It was concluded that a non-exercise model based on BIA might be a rapid and useful technique to estimate VO(2max), when a direct test does not seem feasible. However, though the present results are useful to determine the viability of the method, further refinement of the BIA approach and its validation in a large, diverse population is needed before it can be applied to the clinical and epidemiological settings. PMID- 16261390 TI - Central perineal hernia mimicking rectocele: a case report. AB - BACKGROUND: A perineal hernia is a very rare clinical finding. Three forms are distinguished: anterior, posterior, and central. Diagnosis of the last one is difficult, and sometimes, it is falsely named a posterior rectocele. AIM: This work presents a successfully treated case of central perineal hernia and makes a brief summary of existent literature on the problem. PRESENTATION OF THE CASE: We report of a 67-year-old female patient with a symptomatic central pelvic floor hernia. After radiological confirmation of the diagnosis, a transperitoneal approach was chosen to reposition the protruded segment of the small bowel. The hernial orifice was closed by extraperitoneal implantation of a polypropylene mesh. DISCUSSION: In the present case, the use of a laparoscopic technique seemed unsuitable due to the extension of the findings. For the repair of perineal hernia, we followed the principles of the "tension-free" concept. If there are no signs of a pelvic floor infection and if the mesh can be implanted totally extraperitoneally, we recommend the use of nonabsorbable alloplastic material (polypropylene) for reinforcement of the pelvic floor as a suitable technique for the repair of large perineal hernias. PMID- 16261391 TI - Predictive factors of liver injury in blunt multiple trauma. AB - INTRODUCTION: This study was conducted to clarify whether injuries that are likely to be revealed by initial clinical and conventional radiological examination at the trauma bay (e.g., right-side rib fractures) meaningfully contribute to the prior probability of accompanying hepatic lesions in multiple injured patients. MATERIAL AND METHODS: Fifty-five subjects (sampled from a cohort of 218 patients) with liver injury fulfilling the definition of polytrauma were compared with 55 polytrauma patients without liver injury. Controls were individually matched for age, gender, and Injury Severity Scores. Whole-body, helical, contrast-enhanced computed tomography was applied to all participants. We modeled independent predictors of liver involvement by conditional logistic and random-effects regression analysis. RESULTS: In the present sample, the prevalence of hepatic injury was 25.2%. Neither the injury mechanism (car crash, pedestrian accident, fall from height) nor certain accompanying injuries (right side serial rib fractures, lumbar spine fractures) predicted the presence of hepatic injury. Liver injury was particularly unlikely in bikers [odds ratio (OR) 0.78, 95% confidence interval (CI) 0.59-1.03] and patients with left-side rib fractures (OR 0.80, 95% CI 0.66-0.98). DISCUSSION: There are no index injuries that will reliably indicate the presence of liver involvement in multiple trauma cases. Also, the absence of these injuries cannot rule out liver damage. PMID- 16261392 TI - Virtual placement of posterior C1-C2 transarticular screw fixation. AB - We wanted to evaluate how often safe and effective posterior C1-C2 transarticular screw placement is realizable when it is performed according to guidelines given in the literature. In 50 adult patients, computerized tomography scan data from C0 to C3 were transformed into a 3D spine model. Virtually, bilateral screws were placed from the medial third of the C2-C3 facet joint towards the rim of the C1 anterior arc parallel to midline. Three categories of virtual screw position were rated: optimal (virtual screw inside the C2 pars interarticularis, transversing the middle third of the atlantoaxial joint, and sparing the vertebral artery canal), suboptimal (virtual screw violating the C2 pars interarticularis, and/or transversing the lower or upper third of the C1-C2 joint, and sparing vertebral artery canal), and unacceptable (virtual screw breaching the vertebral artery canal). Optimal placement was seen in 74, suboptimal placement in 11, and unacceptable locations in 15 sites. We conclude that due to the variability of the anatomy of the upper cervical spine, optimal transarticular C1-C2 screw placement is not possible in up to 26%, and even hazardous in up to 15%. PMID- 16261393 TI - [Bacterial spectra and antibiotics in odontogenic infections. Renaissance of the penicillins?]. AB - BACKGROUND: The role played by odontogenic infection in dental, oral, and maxillofacial surgery is not to be underestimated even at the present time. An extensive, standardized, prospective study was performed with the intention of verifying the bacterial spectrum of odontogenic infections to evaluate antibiotic sensitivity. MATERIAL AND METHODS: Bacterial spectra and resistograms of 65 patients with an odontogenic infection were analyzed in a prospective study under standardized conditions for specimen collection and transport. RESULTS: A total of 226 bacterial strains were analyzed. The ratio between anaerobes and aerobes was approximately 2:1. The most frequent aerobes were members of the genera Streptococcus (46 isolates), Staphylococcus (10 isolates), and Neisseria (9 isolates), respectively. The anaerobic gram-positive spectrum was dominated by members of the genera Eubacterium (19 isolates), Peptostreptococcus (16 isolates), and Actinomyces (12 isolates). The most frequently isolated gram negative anaerobes were Prevotella (46 isolates), and Fusobacterium (21 isolates). The overall resistance to antibiotics was very low: only 7.3% of all bacteria were resistant to penicillin G/V, and 8.8% showed resistance to ampicillin. The resistance rates to other beta-lactam antibiotics were 4.4% to piperacillin and 0.6% to imipenem, respectively. Penicillin G presented the highest antimicrobial activity among aerobes: only 4.5% of anaerobic strains were resistant of penicillin G. The other resistance rates of anaerobic bacteria to antibiotics were as follows: ampicillin 24%, doxycycline 34%, erythromycin 18%, and clindamycin 9.3%. Penicillin G was also highly antimicrobially active to anaerobes. The resistance rates were: penicillin G 8.1%, ampicillin 2.6%, doxycycline 9.2%, erythromycin 10.2%, and clindamycin 1.4%, respectively. PMID- 16261394 TI - Preoperative progressive pneumoperitoneum in patients with abdominal-wall hernias. AB - Induction of preoperative progressive pneumoperitoneum is an elective procedure in patients with hernias with loss of domain. A prospective study was carried out from June 2003 to May 2005 at the Hospital de Especialidades, Instituto Mexicano del Seguro Social, Leon, Mexico. Preoperative progressive pneumoperitoneum was induced using a double-lumen intraabdominal catheter inserted through a Veress needle and daily insufflation of ambient air. Variables analyzed were age, sex, body mass index, type, location and size of defective hernia, number of previous repairs, number of days pneumoperitoneum was maintained, type of hernioplasty, and incidence of complications. Of 12 patients, 2 were excluded because it was technically impossible to induce pneumoperitoneum. Of the remaining 10 patients, 60% were female and 40% were male. The patients' average age was 51.5 years, average body mass index was 34.7, and evolution time of their hernias ranged from 8 months to 23 years. Nine patients had ventral hernias and one had an inguinal hernia. Pneumoperitoneum was maintained for an average of 9.3 days and there were no serious complications relating to the puncture or the maintenance of the pneumoperitoneum. One patient who previously had undergone a mastectomy experienced minor complications. We were able to perform hernioplasty on all patients, eight with the Rives technique, one with supra-aponeurotic mesh, and one using the Lichtenstein method for inguinal hernia repair. One patient's wound became infected postoperatively. Preoperative progressive pneumoperitoneum is a safe procedure that is easy to perform and that facilitates surgical hernia repair in patients with hernia with loss of domain. Complications are infrequent, patient tolerability is adequate, and the proposed modification to the puncture technique makes the procedure even safer. PMID- 16261395 TI - Homeward bound: moving treatment from the institution to the community. AB - This study examined changes in the length of stay, cost savings, recidivism and community access when individuals with serious mental illness who were mandated into extended treatment were moved from a regional center institution to community treatment. Results showed significantly shorter length of stay, cost savings and no increase in recidivism when individuals were treated in the community program. PMID- 16261396 TI - Neuregulins 1-4 are expressed in the cytoplasm or nuclei of ductal carcinoma (in situ) of the human breast. AB - A new family of epidermal growth factor-like proteins, the Neuregulins (NRGs), have recently been identified and are expressed in a range of normal tissues and in some forms of cancer including breast cancer. In this study we examined using immunohistochemical staining expression of NRG1alpha, NRG1beta, NRG2alpha, NRG2beta, NRG3 and NRG4 in sixty cases of pre-invasive ductal carcinoma in situ of the breast representing different degrees of differentiation. Each protein was expressed in a high proportion of these cases showing a predominantly homogenous cytoplasmic staining pattern. Nuclear expression of NRG1alpha, NRG1beta, and NRG3 was however also observed in a significant fraction of cases. High levels of expression of NRG2beta and NRG4 were associated with high-grade tumours (p< or =0.005), NRG2beta staining was associated with tumour size >25 mm (p=0.005) while NRG3 nuclear staining was present more often in low-grade tumours (p=0.039). This data demonstrates that each member of the NRG family of ligands is present in pre invasive ductal breast cancer and that they may be involved in regulating cell behaviour. The significance of intranuclear expression remains to be determined but suggests a novel mechanism of action for some of these proteins. PMID- 16261397 TI - Bidirectional cross talk between ERalpha and EGFR signalling pathways regulates tamoxifen-resistant growth. AB - We have previously demonstrated that oestrogen receptor alpha (ERalpha) modulates epidermal growth factor receptor (EGFR)/mitogen-activated protein kinase (MAPK) signalling efficiency in a tamoxifen-resistant MCF-7 breast cancer cell line (Tam R). In the present study we have investigated whether this cross-talk between EGFR/MAPK and ERalpha signalling pathways is bidirectional by examining the effects of EGFR/MAPK activity on ER functionality in the same cell line. Elevated expression levels of phosphorylated serine 118 (S118) ERalpha were observed in the Tam-R compared to the parental wild type MCF-7 cell line (WT-MCF-7) under basal growth conditions. Phosphorylation of ERalpha at S118 was regulated by the EGFR/MAPK pathway in Tam-R cells being increased in response to amphiregulin (AR) and inhibited by the selective EGFR tyrosine kinase inhibitor, gefitinib and the MEK1/2 inhibitor, PD184352. Recruitment of the co-activators p68 RNA helicase and SRC1 to ERalpha, oestrogen response element (ERE) activity and Tam-R cell growth were similarly EGFR/MAPK-regulated. Chromatin immunoprecipitation (ChIP) studies revealed that in Tam-R cells the ERalpha assembled on the AR gene promoter and this was associated with elevated basal expression of AR mRNA. Furthermore, AR mRNA expression was under the regulation of the EGFR/MAPK and ERalpha signalling pathways. Neutralising antibodies to AR inhibited EGFR/ERK1/2 activity, reduced S118 ERalpha phosphorylation and reduced AR mRNA expression in TAM-R cells. These findings suggest that ERalpha function in Tam-R cells is maintained as a consequence of EGFR/MAPK-mediated phosphorylation at serine residue 118 resulting in the generation of a self-propogating autocrine growth-regulatory loop through the ERalpha-mediated production of AR. PMID- 16261398 TI - The (mis)classification of chemo-fog--methodological inconsistencies in the investigation of cognitive impairment after chemotherapy. AB - BACKGROUND: A growing number of studies report cognitive impairment after chemotherapy; indeed the phenomenon of chemo-fog is now almost universally accepted. We are concerned however that there is little if any consistency in the way in which patients are classified as showing cognitive impairment or not. We aim to demonstrate that different methods of analysis produce markedly different results, making the true extent of impairment unclear. METHODS: We analysed data from 92 breast cancer patients 4 weeks post-chemotherapy and from 42 healthy controls using 7 different methods, each taken from a different research paper in the area of cognitive impairment post-chemotherapy. FINDINGS: The extent of impairment was dependent on the method of analysis. Impairment ranged from 12 to 68.5% in the chemotherapy group and from 4.8 to 64.3% in the healthy control group. INTERPRETATION: This brief report highlights the contrasting degrees of cognitive impairment calculated by using legitimate statistical methods and demonstrates the need for a collaborative effort to standardise our methods that we might better understand the phenomenon of chemo-fog. PMID- 16261399 TI - Influence of selected lifestyle factors on breast and ovarian cancer risk in BRCA1 mutation carriers from Poland. AB - It has been estimated that the lifetime risk of breast cancer among women who inherit a BRCA1 or BRCA2 mutation is as high as 80%, and the risk estimates for ovarian cancer range from 15 to 40%. Several environmental and lifestyle factors are believed to contribute to the development of breast cancer in the general population and it is of interest to establish if these factors operate among mutation carriers as well. To evaluate the effects of age of menarche, parity, breast-feeding, oophorectomy and oral contraceptive use, as well as smoking and coffee consumption, on the risks of breast and ovarian cancer, we conducted a matched case-control study of Polish women with BRCA1 mutations. There were 348 breast cancer patients, 150 ovarian cancer patients and similar numbers of age matched controls. BRCA1 carriers with late age of menarche, lower parity and long term breast-feeding were less likely to develop breast cancer. Oral contraceptives protected against ovarian cancer. PMID- 16261400 TI - Incidence of BRCA1 and BRCA2 mutations in 54 Chilean families with breast/ovarian cancer, genotype-phenotype correlations. AB - Our aim was to analyze the incidence of mutations in BRCA1 and BRCA2 genes in 54 families with breast/ovarian cancer. Families were selected from three Institutions following the standard criteria for hereditary breast/ovarian cancer. PCR amplification of all exons was performed, followed by SSCP, heteroduplex, PTT and sequencing analysis. We identified eight truncation mutations, three in the BRCA1 gene and five in the BRCA2 gene. Three of these mutations have not been reported previously by other groups: 308insA in one family, 3936 C>T in two families, for BRCA1, and 4970insTG in one family for BRCA2. In addition two families having Ashkenazi Jewish ancestors present the well known mutations 185delAG and 6174delT. Interestingly, 5 out of 11 families have mutations recurrent in Spanish families. Among the 54 families selected, seven have breast and ovary cancer cases, and only two presented a mutation in BRCA1 or BRCA2 genes. Other cancers as prostate and stomach are frequent among relatives carrying the mutation. Five cases of very early onset (<31 years old) breast cancer were detected. The frequencies of BRCA1 (0.074) and BRCA2 (0.13) mutations in our families is low but similar to the incidence found in other populations, like in Spain. Since is widely known that risk factors that modulate the development of breast cancer such as lifestyle risk factors, geographic location, country of origin and socioeconomic status, besides a familial history of breast cancer our findings suggest that the history of colonization and immigrations is very relevant when studying hereditary factors associated to breast cancer. PMID- 16261401 TI - A French national survey on infiltrating breast cancer: analysis of clinico pathological features and treatment modalities in 1159 patients. AB - BACKGROUND: Despite the approximate 42,000 yearly new cases of breast cancer in France, there have been very few exhaustive studies on the clinicopathological features and treatment options of this disease. METHODS: Thus, a prospective, non selective, nationwide survey on infiltrating breast cancer (IBC) was conducted in France from September 2001 to April 2002, in order to assess the epidemiological features of newly diagnosed disease, the prognostic and predictive variables with a special emphasis on hormone receptors, and the current approaches to therapy in everyday clinical practice. RESULTS: In total, 1159 patients were evaluable (median age 57 years); two-thirds of women were postmenopausal and 38% had undergone hormonal replacement therapy (HRT). Ductal and lobular infiltrating cancers represented 82.3% and 11.6% of cases, respectively. Most tumours expressed oestrogen (79.7%) and progesterone (69.7%) receptors. Overexpression of the human epidermal growth factor receptor-2 oncogene was found in 20.6% of the assessed cases. IBC diagnosed in women under HRT presented significantly better clinico-pathological features than in non-users. All patients underwent surgery as first treatment: 77.5% breast-conserving surgery (BCS) and 22.5% mastectomy; 1024 patients also underwent axillary surgery. The overall axillary lymph-node involvement rate was 44.4%. Radiotherapy was proposed in 98% and 83% of the women who had undergone BCS and mastectomy, respectively. Adjuvant chemotherapy was delivered in 58.7% of patients and hormonal treatment was provided in 76.5% of patients; tamoxifen was the most widely used hormonal treatment. CONCLUSIONS: This study showed a trend for global downstaging of IBC (with favourable clinico pathological features), leading to a high rate of BCS. Postoperative treatments were widely used, in accordance with national and international guidelines. Use of aromatase inhibitors and taxanes was limited, but is likely to rise in the future. PMID- 16261402 TI - Nipple-sparing mastectomy in association with intra operative radiotherapy (ELIOT): A new type of mastectomy for breast cancer treatment. AB - BACKGROUND: Breast-conserving surgery has become the standard approach for about 80% of patients treated for primary breast cancer in most centres. However, mastectomy is still required in case of multicentric and/or large tumours or where recurrences occur after conservative treatment. When a total mastectomy is performed, the removal of the nipple areola complex (NAC) is a strongly debated issue. In fact, although removal of the NAC greatly increases the patient's sensation of mutilation, and the risk of tumor involvement of the areola is reported as a very variable percentage, NAC excision still remains the standard treatment. PATIENTS AND METHODS: From March 2002 to September 2003, 106 nipple sparing mastectomies (NSM) were peformed in 102 patients, 63% of whom had invasive carcinoma and 37% of whom had in situ carcinoma. Four patients underwent bilateral surgery. In all cases, a large or multicentric tumour and/or diffuse microcalcifications, clinically distant from the NAC, were present. During surgery, the tissue under the areola was routinely sampled to exclude the presence of tumor. If disease-free at the frozen sections, the NAC was spared and a NSM was performed. Additionally, a total dose of 16 Gy of radiotherapy (ELIOT) was delivered intraoperatively in the region of the NAC. All the patients underwent an immediate plastic breast reconstruction. RESULTS: In eleven patients (10.4%), the breast tissue under the areola resulted infiltrated at the definitive histological examination: in 10 cases a single or multiple foci of in situ carcinoma and in one case an invasive component were present. Eleven patients (10.4%) developed a superficial skin areolar slough followed by spontaneous healing, and 5 patients (4.7%) lost their NAC due to total necrosis. Among these, one patient had a poor cosmetic result on the NAC with asymmetrical location and required further surgical removal and reconstruction with tattoo and local flap in a better position. When rating the results from 0 (bad) to 10 (excellent), on average, the colour of the areola was rated 9/10, the sensitivity of nipple 3/10, the overall aesthetic result was rated 8/10 by both the surgeon and the patients. Early radiodystrophy (pigmentation) was observed in eight cases (7.5%). After an average follow up of 13 months, one local recurrence, located under the clavicula, far from the NAC, was observed. The preliminary results of the psychological study show a very high satisfaction with the preservation of the nipple (97.6 %), with younger women expressing a higher satisfaction than older counterparts. CONCLUSIONS: In selected cases, NSM with ELIOT of NAC has so far permitted good local control of the disease and satisfactory cosmetic results. Wider surgical experience is required to minimise the risk of leaving tumor cells in the region of the spared NAC and a longer follow up is necessary to evaluate the long term tumor recurrence rate at the NAC. PMID- 16261403 TI - Clinical response to neoadjuvant docetaxel predicts improved outcome in patients with large locally advanced breast cancers. AB - PURPOSE: In the adjuvant setting, taxanes modestly improve clinical outcome and survival. The goal of the present study was to define the efficacy of neoadjuvant docetaxel in treatment-naive large, locally advanced breast cancers and to better understand docetaxel's mechanism of action by evaluating biomarker modulation in response to treatment. PATIENTS AND METHODS: Fifty-one patients were enrolled. Patients received four cycles of docetaxel (100 mg/m2 q3weeks) followed by surgery and four cycles of doxorubicin and cyclophosphamide (60/600 mg/m2 q3weeks). Radiation and hormonal therapy were given if clinically indicated. Clinical responses were assessed at completion of neoadjuvant docetaxel. Pathological responses were considered complete (pCR) if no tumor cells were identified in the surgical specimen or near complete (npCR) if only occasional scattered tumor cells were seen. Proliferation (Ki-67) and apoptosis (cleaved caspase-3) were measured by IHC in tissue obtained at baseline and at surgery. RESULTS: The median tumor size was 9 cm (range 4-30 cm). Objective response rate was 75% with clinical complete response in 27%, partial response in 48%, and stable disease in 25% of the patients. pCR/npCR was reported in 20% of patients. With a median follow up of 28 months, 98 and 78% of the patients were alive at 12 and 24 months, respectively. Overall survival at 24 months was significantly better in patients who achieved a clinical response, 85 versus 51%, p = 0.008, but pCR/npCR was not a significant predictor of outcome. Apoptosis was induced in clinical responders (p = 0.002), while the proliferation index did not change significantly. In patients who had no clinical response to docetaxel, neither apoptosis nor proliferation changed significantly. CONCLUSION: Neoadjuvant single agent docetaxel is effective in treating patients with large locally advanced breast cancer and clinical response is associated with improved survival. Docetaxel acts therapeutically by inducing apoptosis and this can be used as a marker of response. PMID- 16261404 TI - Identification of cell-of-origin breast tumor subtypes in inflammatory breast cancer by gene expression profiling. AB - Inflammatory breast cancer (IBC) is an aggressive form of locally advanced breast cancer with high metastatic potential. Most patients have lymph node involvement at the time of diagnosis and 1/3 of the patients have distant metastases. In a previous study, we demonstrated that IBC is a distinct form of breast cancer in comparison with non-IBC. The aim of this study was to investigate the presence of the different molecular subtypes in our data set of 16 IBC and 18 non-IBC specimen. Therefore, we selected an 'intrinsic gene set' of 144 genes, present on our cDNA chips and common to the 'intrinsic gene set' described by Sorlie et al. [PNAS, 2003]. This set of genes was tested for performance in the Norway/Stanford data set by unsupervised hierarchical clustering. Expression centroids were then calculated for the core members of each of the five subclasses in the Norway/Stanford data set and used to classify our own specimens by calculating Spearman correlations between each sample and each centroid. We identified the same cell-of-origin subtypes in IBC as those already described in non-IBC. The classification was in good agreement with immunohistochemical data for estrogen receptor protein expression and cytokeratin 5/6 protein expression. Confirmation was done by an alternative unsupervised hierarchical clustering method. The robustness of this classification was assessed by an unsupervised hierarchical clustering with an alternative gene set of 141 genes related to the cell-of origin subtypes, selected using a discriminating score and iterative random permutation testing. The contribution of the different cell-of-origin subtypes to the IBC phenotype was investigated by principal component analysis. Generally, the combined ErbB2-overexpressing and basal-like cluster was more expressed in IBC compared to non-IBC, whereas the combined luminal A, luminal B and normal like cluster was more pronounced in non-IBC compared to IBC. The presence of the same molecular cell-of-origin subtypes in IBC as in non-IBC does not exclude the specific molecular nature of IBC, since gene lists that characterize IBC and non IBC are entirely different from gene lists that define the different cell-of origin subtypes, as evidenced by principal component analysis. PMID- 16261405 TI - The time-related changes of the importance of prognostic factors in breast cancer. A sequential multivariate analysis of 1423 Japanese patients. AB - The value of individual prognostic factors may change dependent on the length of the follow-up period, if some factors have their greatest prognostic potential immediately after operation. It is not clear how long these factors keep their prognostic relevance. We retrospectively examined data on 1423 surgically treated Japanese patients with primary breast cancer between 1983 and 2002. Survival analysis was done starting at 2.5-yearly intervals after operation and follow-up in the first analysis started at the time of the operation. The changing importance of the prognostic factors during different follow-up periods was investigated by univariate and multivariate analysis. Based on multivariate analysis, tumor size retained its prognostic value even up to 7.5 years after operation, whereas the age, vascular involvement, ER and PgR showed a changing influence on prognosis dependent on the length of the follow-up period. The prognosis of patients some years after operation is necessarily different from the initial prognosis established after operation. Detecting the changing importance of prognostic factors could provide new biological insights that might otherwise be missed, and may help determine the most appropriate clinical use of various factors. PMID- 16261406 TI - EGFR signaling pathway in breast cancers: from traditional signal transduction to direct nuclear translocalization. AB - Aberrant epidermal growth factor receptor (EGFR) signaling is a major characteristic of many human malignancies including breast cancer. Since the discovery of EGF in 1960's and its receptor in 1980's, our understanding of the EGF/EGFR pathway has been significantly advanced and consequently, EGFR is considered as a major oncogenic factor and an attractive therapeutic target. The well-established traditional function of EGFR is known to transmit extra-cellular mitogenic signals, such as EGF and transforming growth factor-alpha (TGF-alpha), through activating a number of downstream signaling cascades. These include signaling modules that involve phospholipase C-gamma, Ras, and phosphatidylinositol-3 kinase (PI-3K). In cancer cells, the common outcomes following the activation of the EGFR-mediated downstream pathways are altered gene activities, leading to un-controlled tumor proliferation and apoptosis. Interestingly, emerging evidences suggest the existence of a direct mode of the EGFR pathway that is distinct from the traditional transduction pathway. This new mode of EGFR signaling involves cellular transport of EGFR from the cell-surface to the cell nucleus, association of nuclear EGFR complex with gene promoters, and transcriptional regulation of the target genes. Although the nature and pathological consequences of the nuclear EGFR pathway remain elusive, accumulating evidences suggest its association with increased tumor cell proliferation and poor survival rate in breast cancer patients. While several anti-EGFR agents are being tested in breast cancer patients clinically and others under pre-clinical development, a better understanding of the traditional and the nuclear EGFR pathways will facilitate the identification of patients that are likely to respond to these agents as well as future development of more effective anti-EGFR therapeutic interventions. PMID- 16261407 TI - Is adjuvant therapy for older patients with node (+) early breast cancer cost effective? AB - BACKGROUND: Node (+) breast cancer represents over 40% of cases in older women and currently there is a debate whether adjuvant therapy for all older women is cost-effective. PURPOSE: To evaluate if adjuvant treatment for early-stage (Stage I-IIIa) node (+) breast cancer with hormone therapy, chemotherapy, or combination therapy is cost-effective in older patients. DESIGN: A decision-analysis model for 65, 75, and 85 year-old female breast cancer patients using life tables integrated the cost of treatment in dollars and impact in length and quality of life. Both estrogen receptor (ER) (-) and (+) patients were considered. The primary data sources were meta-analysis from the Early Breast Cancer Trialists' Collaborative Group and the Red Book Average Wholesale Price for drugs. The cost of treatment in dollars and impact of quality of life was examined. Scenarios were used when treatment benefit was uncertain. The incremental cost effectiveness of different treatment strategies were then compared and mapped graphically. RESULTS: Adjuvant therapy is cost-effective in 65 year-old women with early breast cancer. In a 75 year-old ER (+) patient, hormone therapy is cost-effective, $10,965/quality-adjusted life years (QALY), but chemotherapy was more cost-effective, $27,406/QALY, if one assumed it was as efficacious as in a 65 year-old woman. In a 75 year-old ER (-) patient, chemotherapy was cost effective at $42,605 with the same assumption. In an 85 year-old ER (+) patient, hormone therapy was cost-effective, $26,463/QALY, if efficacy is not age sensitive, but chemotherapy was not as cost-effective for either ER (+) or ER (-) patients. CONCLUSION: Treatment decisions for older breast cancer patients suffer from the lack of sufficient clinical trial data. Decision-analytic models can help policy makers who are faced with decisions about whether to support adjuvant therapy in older breast cancer patients and also outline the important parameters that need to be considered in such a decision. PMID- 16261408 TI - Polymorphisms of the BRCA2 and RAD51 genes in breast cancer. AB - We performed a case-control study (150 cases and 150 controls) to test the association between three polymorphisms in BRCA2 and RAD51 genes and breast cancer risk. Genotypes were determined in DNA from blood cells by PCR-RFLP. Cancer occurrence was strongly associated with the BRCA2 Met/1915Thr homozygous polymorphic variants, whereas heterozygous variant was associated with significant reduction in breast cancer risk. Gene-gene interaction between the BRCA2-Met1915Thr Thr/Thr and BRCA2-Met784Val Met/Met homozygous variants increased the risk. Therefore, the Met1915Thr polymorphism in the BRCA2 gene may be considered as an independent marker of breast cancer. PMID- 16261409 TI - High incidence of 4153delA BRCA1 gene mutations in Lithuanian breast- and breast ovarian cancer families. AB - BRCA1 and BRCA2 gene mutations confer a high lifetime risk to breast and ovarian cancers. We have screened cancer patients from 13 families with at least three breast and/or ovarian cancers from Lithuania for 5382insC, C61G and 4153delA BRCA1 gene mutations. One of three mutations was found in 9 of the 13 studied families (69%). 4153delA was the most frequently detected and accounted for 56% of all identified mutation. 5382insC and C61G accounted for 33% and 11% of found mutations, respectively. Significantly higher, than in other populations, incidence of 4153delA indicates that this may be founder BRCA1 mutation characteristic for Lithuanians. Our analysis shows that testing of 4153delA, 5382insC, C61G BRCA1 mutations should be extremely effective and inexpensive tool in testing Lithuanian population aimed to identify individuals with high risk of breast and ovarian cancers. PMID- 16261410 TI - Mammographic breast density and the Gail model for breast cancer risk prediction in a screening population. AB - BACKGROUND: Estimating an individual woman's absolute risk for breast cancer is essential for decision making about screening and preventive recommendations. Although the current standard, the Gail model, is well calibrated in populations, it performs poorly for individuals. Mammographic breast density (BD) may improve the predictive accuracy of the Gail model. METHODS: Prospective observational cohort of 81,777 women in the San Francisco Mammography Registry presenting for mammography during 1993 through 2002 who had no prior diagnosis of breast cancer. Breast density was rated by clinical radiologists using the Breast Imaging Reporting and Data System classification (almost entirely fat; scattered fibroglandular densities; heterogeneously dense; extremely dense). Breast cancer cases were identified through linkage to Northern California Surveillance Epidemiology End Results (SEER) program. We compared the predictive accuracy of models with Gail risk, breast density, and the combination. All models were adjusted for age and ethnicity. RESULTS: During 5.1 years of follow-up, 955 women were diagnosed with invasive breast cancer. The Gail model had modest predictive accuracy (concordance index (c-index) 0.67; 95% CI 0.65-0.68). Adding breast density to the model increased the predictive accuracy to 0.68 (95% CI .66-.70, p < 0.01 compared with the Gail model alone). The model containing only breast density adjusted for age and ethnicity had predictive accuracy equivalent to the Gail model (c-index 0.67, 95% CI 0.65-0.68). CONCLUSION: The addition of breast density measured by BI-RADS categories minimally improved the predictive accuracy of the Gail model. A model based on breast density alone adjusted for age and ethnicity was as accurate as the Gail model. PMID- 16261411 TI - A cost-effectiveness analysis of two rehabilitation support services for women with breast cancer. AB - The purpose of this research was to estimate the cost-effectiveness of two rehabilitation interventions for breast cancer survivors, each compared to a population-based, non-intervention group (n = 208). The two services included an early home-based physiotherapy intervention (DAART, n = 36) and a group-based exercise and psychosocial intervention (STRETCH, n = 31). A societal perspective was taken and costs were included as those incurred by the health care system, the survivors and community. Health outcomes included: (a) 'rehabilitated cases' based on changes in health-related quality of life between 6 and 12 months post diagnosis, using the Functional Assessment of Cancer Therapy-Breast Cancer plus Arm Morbidity (FACT-B+4) questionnaire, and (b) quality-adjusted life years (QALYs) using utility scores from the Subjective Health Estimation (SHE) scale. Data were collected using self-reported questionnaires, medical records and program budgets. A Monte-Carlo modelling approach was used to test for uncertainty in cost and outcome estimates. The proportion of rehabilitated cases was similar across the three groups. From a societal perspective compared with the non-intervention group, the DAART intervention appeared to be the most efficient option with an incremental cost of $1344 per QALY gained, whereas the incremental cost per QALY gained from the STRETCH program was $14,478. Both DAART and STRETCH are low-cost, low-technological health promoting programs representing excellent public health investments. PMID- 16261412 TI - Serum lipids and outcome of early-stage breast cancer: results of a prospective cohort study. AB - PURPOSE: The prognosis of women with early-stage breast cancer is influenced by insulin and body mass index (BMI). High levels of serum insulin and obesity often coexist with dyslipidemia in the insulin resistance syndrome (IRS), but the contribution of lipids to breast cancer outcome is unclear. Here, we examine whether serum levels of total cholesterol (TC) and triglycerides (TG) influence breast cancer outcome. PATIENTS AND METHODS: A cohort of 520 women without known hyperlipidemia or diabetes, with stage T1-T3, N0-N1, M0 breast cancer, was assembled from July 1989 to June 1996. Fasting blood was collected at baseline. Subjects were followed prospectively, for recurrence (local, regional, distant) and death. Cox models were used to calculate the prognostic effect of TC and TG levels. Two-sided significance levels were set at 0.025. RESULTS: TC was correlated with age (Spearman's r = 0.44) and low tumor grade (p = 0.01), while TG was correlated with insulin (r = 0.43) and BMI (r = 0.45). At a median follow up of 8.7 years, TC and TG were not associated with breast cancer recurrence or death before of after adjustment for age, tumor-related variables, BMI or fasting insulin levels. In multivariate analysis adjusting for age, tumor-related variables and BMI, a trend towards an adverse effect of TC on disease recurrence was seen (HR recurrence = 1.62 for the 4th versus. 1st quartile, 2-sided p = 0.03). CONCLUSIONS: Fasting TG was not associated with outcome. A trend towards risk of recurrence was seen with higher TC in multivariate analysis. This potential association should be explored in future studies. PMID- 16261413 TI - Estrogen receptor beta (ESR2) polymorphisms in familial and sporadic breast cancer. AB - Estrogen is involved in both normal mammary development and in breast carcinogenesis. A family history of disease and exposure to estrogen are major risk factors for developing breast cancer. Estrogen exerts its biological effects through binding to the estrogen receptors, estrogen receptor alpha (ESR1) and the more recently discovered estrogen receptor beta (ESR2). Genetic variation in genes involved in estrogen biosynthesis, metabolism and signal transduction have been suggested to play a role in breast cancer risk. We therefore tested the hypothesis that common genetic variants of the ESR2 gene may be associated with increased risk for breast cancer and this risk may vary between breast cancer groups. We investigated three common ESR2 polymorphisms, rs1256049 (G1082A), rs4986938 (G1730A) and rs928554 (Cx+56 A-->G) for association to breast cancer risk. A total of 723 breast cancer cases and 480 controls were included in the study. Of the breast cancer cases, 323 were sporadic and 400 were familial, the familial cases were further divided into familial high-risk and familial low-risk breast cancer cases. We found no overall statistically significant association for any of the single polymorphisms studied. Haplotype analysis suggested one haplotype associated with increased risk in sporadic breast cancer patients (OR = 3.0, p = 0.03). Further analysis is needed to elucidate the role of estrogen receptor beta in breast cancer susceptibility. PMID- 16261414 TI - How should we inform women at higher risk of breast cancer about tamoxifen? An approach with a decision guide. AB - BACKGROUND: Tamoxifen has been shown to reduce the incidence of invasive breast cancer in women at higher risk. Translating these research results to clinical practice is challenging. Our objective was to develop and evaluate a decision making guide and process that can be used in clinical practice to inform eligible women of chemoprevention with tamoxifen. METHODS: A decision guide explaining the benefits and risks of tamoxifen was developed with input from health care professionals and two focus groups of women both with and without cancer. Following consent, 51 eligible women presenting to a multi-disciplinary diagnostic facility for breast problems were given the decision guide/questionnaire to read, fill out and return by mail. Women with further questions or wanting to take tamoxifen were encouraged to re-contact their physicians. RESULTS: Atypia was seen in 60% of subjects. Median 5-year Gail risk was 3.7 (range 1.7-9.4). Only 6 (11.8% 95% CI = 2.9, 20.6%) women reported they would like to take tamoxifen while 6 (11.8% 95% CI = 2.9, 20.6%) remained uncertain. CONCLUSION: We have developed a decision-making guide and process that is acceptable to providers and women to identify and inform women at higher risk of breast cancer with regard to chemoprevention with tamoxifen. Few women in this select group, when provided with a balanced decision guide, wished to pursue chemoprevention with tamoxifen. PMID- 16261415 TI - Anti-estrogenic effects of conjugated linoleic acid through modulation of estrogen receptor phosphorylation. AB - We previously showed that conjugated linoleic acids (CLA) can inhibit transcriptional activation mediated by estrogen response elements (EREs) and that this activity can, at least in part, account for the reported anti-tumor effects of these compounds on breast cancer cells. Using estrogen receptor positive (ER+) MCF-7 cells, we now demonstrate that CLA inhibited both the transactivation of artificial reporter constructs driven by canonical EREs, and the expression of endogenous progesterone receptors, a gene which is transcriptionally regulated by estrogen through novel ER-binding sites. This inhibition was accompanied by downregulation of ER alpha expression and decreased ER alpha-ERE binding activity. These effects on ER alpha were not causally linked since transfection of an ER alpha expression plasmid in MCF-7 cells failed to antagonize CLA downregulation of ER alpha-ERE binding. Immunoprecipitation/Western blot studies revealed that CLA dose-dependently suppressed the degree of phosphorylation of ER alpha, a modification known to inhibit receptor-ERE interactions. As a mechanism that may account for this induced dephosphorylation of ER alpha in MCF-7, we found that CLA specifically stimulated protein phosphatase 2A (PP2A) activity. Experiments using the PP2A inhibitor okadaic acid (OA) showed that OA antagonized both the dephosphorylation effects of CLA on ER alpha and its inhibition of ER alpha-ERE binding. These results provide evidence that the anti-estrogenic activity of CLA is caused by inducing the dephosphorylation of ER alpha through stimulation of PP2A activity. PMID- 16261416 TI - The influence of muscle type and dystrophin deficiency on murine expression profiles. AB - The phenotypic differences among Duchenne muscular dystrophy patients, mdx mice, and mdx(5cv) mice suggest that despite the common etiology of dystrophin deficiency, secondary mechanisms have a substantial influence on phenotypic severity. The differential response of various skeletal muscles to dystrophin deficiency supports this hypothesis. To explore these differences, gene expression profiles were generated from duplicate RNA targets extracted from six different skeletal muscles (diaphragm, soleus, gastrocnemius, quadriceps, tibialis anterior, and extensor digitorum longus) from wild-type, mdx, and mdx(5cv) mice, resulting in 36 data sets for 18 muscle samples. The data sets were compared in three different ways: (1) among wild-type samples only, (2) among all 36 data sets, and (3) between strains for each muscle type. The molecular profiles of soleus and diaphragm separate significantly from the other four muscle types and from each other. Fiber-type proportions can explain some of these differences. These variations in wild-type gene expression profiles may also reflect biomechanical differences known to exist among skeletal muscles. Further exploration of the genes that most distinguish these muscles may help explain the origins of the biomechanical differences and the reasons why some muscles are more resistant than others to dystrophin deficiency. PMID- 16261417 TI - Quantitative trait loci for baseline white blood cell count, platelet count, and mean platelet volume. AB - A substantial genetic contribution to baseline peripheral blood counts has been established. We performed quantitative trait locus/loci (QTL) analyses to identify chromosome (Chr) regions harboring genes influencing the baseline white blood cell (WBC) count, platelet (Plt) count, and mean platelet volume (MPV) in F(2) intercrosses between NZW/LacJ, SM/J, and C57BLKS/J inbred mice. We identified six significant WBC QTL: Wbcq1 (peak LOD score at 38 cM, Chr 1), Wbcq2 (42 cM, Chr 3), Wbcq3 (0 cM, Chr 15), Wbcq4 (58 cM, Chr 1), Wbcq5 (82 cM, Chr 1), and Wbcq6 (8 cM, Chr 14). Three significant Plt QTL were identified: Pltq1 (24 cM, Chr 2), Pltq2 (36 cM, Chr 7), and Pltq3 (10 cM, Chr 12). Two significant MPV QTL were identified, Mpvq1 (62 cM, Chr 15) and Mpvq2 (44 cM, Chr 8). In total, the WBC QTL accounted for up to 31% of the total variance in baseline WBC count, while the Plt and MPV QTL accounted for up to 30% and 49% of the total variance, respectively. These analyses underscore the genetic complexity underlying these traits in normal populations and provide the basis for future studies to identify novel genes involved in the regulation of mammalian hematopoiesis. PMID- 16261418 TI - Genetics of body weight in the LXS recombinant inbred mouse strains. AB - This is the first phenotypic analysis of 75 new recombinant inbred (RI) strains derived from ILS and ISS progenitors. We analyzed body weight in two independent cohorts of female mice at various ages and in males at 60 days. Body weight is a complex trait which has been mapped in numerous crosses in rodents. The LXS RI strains displayed a large range of weights, transgressing those of the inbred progenitors, supporting the utility of this large panel for mapping traits not selected in the progenitors. Numerous QTLs for body weight mapped in single- and multilocus scans. We assessed replication between these and previously reported QTLs based on overlapping confidence intervals of published QTLs for body weight at 60 days and used meta-analyses to determine combined p values for three QTL regions located on Chromosomes 4, 5, and 11. Strain distribution patterns of microsatellite marker genotypes, weight, and other phenotypes are available on WebQTL (http://www.webqtl.org/search.html ) and allow genetic mapping of any heritable quantitative phenotype measured in these strains. We report one such analysis, correlating brain and body weights. Large reference panels of RI strains, such as the LXS, are invaluable for identifying genetic correlations, GXE (Gene X Environment) interactions, and replicating previously identified QTLs. PMID- 16261419 TI - "Agouti NOD": identification of a CBA-derived Idd locus on Chromosome 7 and its use for chimera production with NOD embryonic stem cells. AB - Penetrance of the complex of genes predisposing the nonobese diabetic (NOD) mouse to autoimmune diabetes is affected by the maternal environment. NOD.CBALs Tyr(+)/Lt is an agouti-pigmented Chromosome 7 congenic stock of NOD/Lt mice produced as a resource for embryo transfer experiments to provide the necessary maternal factors and allow the easy identification of NOD (albino) embryo donor phenotype. CBcNO6/Lt, a recombinant congenic agouti stock already containing approximately 50% NOD genome, was used as the donor source of a wild-type CBA tyrosinase allele. When the incidence of diabetes was assessed after nine generations of backcrossing and one generation of sib-sib mating, significant reduction in diabetes development was observed. No difference in diabetes development was observed in Tyr/Tyr(c) heterozygotes, showing that protection was recessive. Analysis of diabetes progression in another NOD stock congenic for C57BL/6 alleles on Chromosome 7 linked to the glucose phosphate isomerase (Gpi1(b)) locus provided no protection, indicating that the diabetes resistance (Idd) gene was distal to 34 cM (D7Mit346). Approximately 5 cM of the distal congenic region overlaps a region from C57L previously associated with protection when homozygous. The delayed onset and reduced frequency of diabetes in the NOD.CBALs-Tyr(+)/Lt stock is an advantage when females of this stock are used as surrogate mothers in studies involving hysterectomy or embryo transfers. Indeed, a newly developed NOD embryonic stem (ES) cell line injected into NOD.CBALs- Tyr(+)/Lt blastocysts produced approximately 50% live-born mice, of which approximately 11% were chimeric. Presumably because of high genomic instability, no germline transmission was observed. PMID- 16261420 TI - A new method for identifying informative genetic markers in selectively bred rats. AB - Microsatellite length polymorphisms are useful for the mapping of heritable traits in rats. Over 4000 such microsatellites have been characterized for 48 inbred rat strains and used successfully to map phenotypes that differ between strains. At present, however, it is difficult to use this microsatellite database for mapping phenotypes in selectively bred rats of unknown genotype derived from outbred populations because it is not immediately obvious which markers might differ between strains and be informative. We predicted that markers represented by many alleles among the known inbred rat strains would also be most likely to differ between selectively bred strains derived from outbred populations. Here we describe the development and successful application of a new genotyping tool (HUMMER) that assigns "heterozygosity" (Het) and "uncertainty" (Unc) scores to each microsatellite marker that corresponds to its degree of heterozygosity among the 48 genotyped inbred strains. We tested the efficiency of HUMMER on two rat strains that were selectively bred from an outbred Sprague-Dawley stock for either high or low activity in the forced swim test (SwHi rats and SwLo rats, respectively). We found that the markers with high Het and Unc scores allowed the efficient selection of markers that differed between SwHi and SwLo rats, while markers with low Het and Unc scores typically identified markers that did not differ between strains. Thus, picking markers based on Het and Unc scores is a valuable method for identifying informative microsatellite markers in selectively bred rodent strains derived from outbred populations. PMID- 16261421 TI - A QTL resource and comparison tool for pigs: PigQTLDB. AB - During the past decade, efforts to map quantitative trait loci (QTL) in pigs have resulted in hundreds of QTL being reported for growth, meat quality, reproduction, disease resistance, and other traits. It is a challenge to locate, interpret, and compare QTL results from different studies. We have developed a pig QTL database (PigQTLdb) that integrates available pig QTL data in the public domain, thus, facilitating the use of this QTL data in future studies. We also developed a pig trait classification system to standardize names of traits and to simplify organization and searching of the trait data. These steps made it possible to compare primary data from diverse sources and methods. We used existing pig map databases and other publicly available data resources (such as PubMed) to avoid redundant developmental work. The PigQTLdb was also designed to include data representing major genes and markers associated with a large effect on economically important traits. To date, over 790 QTL from 73 publications have been curated into the database. Those QTL cover more than 300 different traits. The data have been submitted to the Entrez Gene and the Map Viewer resources at NCBI, where the information about markers was matched to marker records in NCBI's UniSTS database. Having these data in a public resource like NCBI allows regularly updated automatic matching of markers to public sequence data by e-PCR. The submitted data, and the results of these calculations, are retrievable from NCBI via Entrez Gene, Map Viewer, and UniSTS. Efforts were undertaken to improve the integrated functional genomics resources for pigs. PMID- 16261422 TI - BEGAIN: a novel imprinted gene that generates paternally expressed transcripts in a tissue- and promoter-specific manner in sheep. AB - In this article we describe the organization of the ovine BEGAIN gene, located 138 kb proximally from the imprinted DLK1 gene and 203 kb from the CLPG mutation that causes the callipyge phenotype. We have shown that in sheep BEGAIN is ubiquitously expressed, including in skeletal muscle, throughout development. We have identified four major BEGAIN transcripts resulting from a combination of alternate promoter usage and alternative splicing. In ovine brain, kidney, liver, and skeletal muscle, these four BEGAIN transcripts exhibited paternal or biallelic expression in a tissue- and promoter-specific manner. Our results indicate that the CLPG mutation does not alter transcript levels of BEGAIN, contrary to its effect on a core cluster of genes in the DLK1-GTL2 domain. Thus, although the BEGAIN gene represents another paternally expressed gene in the ovine DLK1-GTL2 imprinted domain, its expression is not governed by the long range regulatory element that contains the CLPG mutation. PMID- 16261423 TI - Isolation and characterization of the leucine-rich proteoglycan nyctalopin gene (cNyx) from chick. AB - We describe the isolation and molecular characterization of the chick ortholog of nyctalopin (NYX), the gene responsible for X-linked complete congenital stationary night blindness (CSNB1, also known as cCSNB). Chick Nyx (cNyx) comprises four exons spanning approximately 6.2 kb on Chromosome 1 and encodes a protein of 473 amino acids that shares 55% identity overall with its human counterpart. cNyx is expressed in both the developing and the fully differentiated retina. Transcripts are localized primarily to cells within the outer half of the inner nuclear layer (INL) and the ganglion cell layer (GCL), a pattern consistent with the principal electrophysiologic findings in CSNB1 that suggest a main defect in depolarizing ON-bipolar cells normally located in the outer half of the INL. Expression (albeit weaker) was also detected in the cerebrum and cerebellum and in non-neuronal tissues. Finally, we also report the identification of three novel splice variants, one of which predominates in the retina. PMID- 16261425 TI - Small animal imaging center design: the facility at the UCLA Crump Institute for Molecular Imaging. AB - PURPOSE: The growing number of mouse and rat experiments, coupled with advances in small-animal imaging systems such as microPET, optical, microCAT, microMR, ultrasound and microSPECT, has necessitated a common technical center for imaging small animals. PROCEDURES: At the UCLA Crump Institute for Molecular Imaging, we have designed and built a facility to support the research interests of a wide range of investigators from multiple disciplines. Requirements to satisfy both research and regulatory oversight have been critically examined. Support is provided for investigator training, study scheduling, data acquisition, archiving, image display, and analysis. RESULTS: The center has been in operation for more than 18 months, supporting more than 13,000 individual imaging procedures. CONCLUSIONS: We have created a facility that maximizes our resource utilization while providing optimal investigator support, as well as the means to continually improve the quality and diversity of the science by integrating physical and biological sciences. PMID- 16261426 TI - Prevalence of severe pelvic organ prolapse in relation to job description and socioeconomic status: a multicenter cross-sectional study. AB - The aim of this study was to determine if certain occupations or socioeconomic levels are associated with pelvic organ prolapse. Investigators at six American sites performed pelvic organ prolapse quantification examinations on women presenting for routine gynecologic care. Between September 1999 and March 2002, 1,004 patients were examined. Severe pelvic organ prolapse was defined as the leading edge being 1 cm or more beyond the hymeneal ring. The data was analyzed with the Kruskal-Wallis analysis of variance, Bonferroni test, multiple logistic regression, and descriptive statistics. The prevalence of severe pelvic organ prolapse in our group was 4.3%. Women who were laborers/factory workers had significantly more severe prolapse than the other job categories (p < 0.001). Women with annual income of Dollars 10,000 or less had significantly more severe pelvic organ prolapse than other income groups (p < 0.001). These differences persisted even when controlling for age, race, number of deliveries, body mass index >30, and smoking status (all p < 0.001). Laborers/factory worker jobs and an annual household income of Dollars 10,000 or less are associated with severe pelvic organ prolapse. PMID- 16261427 TI - Urethral syndrome: response to alpha-adrenergic blocking agents. AB - Urethral syndrome patients have irritative bladder symptoms in the absence of any objective urological findings. While not life threatening, it can have a severe disruptive effect on the quality of life. Attempts at symptom control are not always effective because of lack of clarity about the underlying aetiology. We describe a case of a 27-year-old patient whose irritative bladder symptoms showed response to alpha-adrenergic blocking agents on two separate occasions during the course of her treatment. PMID- 16261428 TI - Mucosal repair and growth factors: recombinant human hepatocyte growth factor as an innovative therapy for inflammatory bowel disease. AB - The repair of intestinal mucosal injuries is a tightly regulated process involving epithelial restitution, cell proliferation and maturation, and the dedifferentiation of epithelial cells. Deeper injuries also require additional repair mechanisms, including inflammatory processes, angiogenesis, and extracellular-matrix deposition. Once intestinal mucosal injury occurs, numerous growth factors and cytokines, including hepatocyte growth factor (HGF), keratinocyte growth factor, endothelial growth factor, epidermal growth factor, transforming growth factor-beta1, intestinal trefoil factor, interleukin (IL)-1, and IL-2, are induced in both the intestinal lumen and submucosa, and these factors cooperatively stimulate epithelial mucosal repair. HGF, a major agent promoting hepatocyte proliferation, also modulates intestinal epithelial cell proliferation and migration, leading to the acceleration of intestinal mucosal repair. Additionally, the proteolytic activation of HGF, which is mediated by HGF activator, is essential for the regeneration of injured intestinal mucosa. Recently, several studies have shown that the administration of recombinant human HGF or HGF gene therapy abrogates disease severity in several animal models of inflammatory bowel disease (IBD). Recombinant human HGF will soon be available for administration to patients with fulminant hepatic failure. Although additional preclinical biological studies are required, HGF has the potential to be an important new treatment modality promoting intestinal mucosal repair in patients with IBD. PMID- 16261429 TI - Self-expandable metallic stent placement for palliation in gastric outlet obstructions caused by gastric cancer: a comparison with surgical gastrojejunostomy. AB - BACKGROUND: In patients with gastric outlet obstruction (GOO), palliative enteral stenting is a less invasive procedure compared with gastroenterostomy. Most diseases analyzed in previous studies of such stenting were pancreaticobiliary malignancies. METHODS: We reviewed the medical records of patients with GOO secondary to gastric cancer who were admitted to our institution between September 1994 and September 2004. The outcome of stent placement for GOO was compared with the outcome in patients who underwent palliative open gastrojejunostomy during the same period. Enrolled patients from both groups displayed symptomatic GOO. Patients with recurrent gastric cancer were excluded from this study. RESULTS: Twenty-two patients underwent palliative enteral stenting, and 22 patients were subjected to surgical gastrojejunostomy (bypass). There were no significant differences between the two groups regarding patient baseline characteristics. Technical success and clinical success were obtained in 100% and 77.3%, respectively, of both groups. The operating time was shorter in the stent group (30 vs 118 min; P<0.0001). The time from the procedure to the resumption of food intake was shorter in the stent group than in the bypass group (2 days vs 8 days; P<0.0001). An improvement in performance score after the procedure was observed in both groups (stent group; P=0.0264; bypass group; P=0.0235). No significant differences were observed regarding the possibility of discharge. In patients discharged, the median postoperative hospital stays were 19 days and 28 days (P=0.0558). The median survival periods were 65 days and 90 days. Minor complications were observed in 1 patient in the stent group and in 4 in the bypass group. No mortality or severe complications were observed for either group. CONCLUSIONS: Self-expandable metallic stent placement is a safe and efficacious procedure for palliation, with shorter operating time and more prompt restoration of oral intake, compared to surgical alternatives in patients with GOO caused by gastric cancer. PMID- 16261430 TI - Serum pepsinogen concentrations as a measure of gastric acid secretion in Helicobacter pylori-negative and -positive Japanese subjects. AB - BACKGROUND: Although previous studies have indicated that serum pepsinogen I levels, as well as the pepsinogen I/II ratio, were positively correlated with maximal gastric output, the relationship may be different between Helicobacter pylori-negative and -positive subjects. The aim of this study was to investigate the relation between serum pepsinogen concentrations and gastric acid secretion in H. pylori-positive and -negative subjects separately. METHODS: The presence of H. pylori infection, the serum pepsinogen concentrations, and gastric acid secretion were investigated in 182 subjects without localized lesions in the upper gastrointestinal tract. Serum pepsinogen concentration was measured by radioimmunoassay, and maximal gastric acid output was estimated by an endoscopic gastrin test, as we have previously shown. RESULTS: In H. pylori-positive subjects, serum pepsinogen I levels and the pepsinogen I/II ratio were significantly correlated with gastric acid secretion, although the latter showed a better correlation (r=0.40 and 0.53, respectively). On the other hand, in H. pylori-negative subjects, serum pepsinogen concentrations were well correlated with acid secretion (r=0.57), but there was no relation between the pepsinogen I/II ratio and acid secretion. CONCLUSIONS: The correlations between serum pepsinogens and gastric acid secretion differ, depending on the presence or absence of H. pylori infection. With the use of serum pepsinogens as a simple measure of gastric acid secretion, therefore, consideration of H. pylori infection status is needed. Because the determination of the acid secretory level has some clinical implications in both H. pylori-positive and -negative subjects, its estimation by serum pepsinogen concentrations can be of practical use. PMID- 16261431 TI - A novel apoptosis-inducing monoclonal antibody (anti-LHK) against a cell surface antigen on colon cancer cells. AB - BACKGROUND: Apoptosis is a crucial element in the behavior of mammalian cells in many different situations. We here report the establishment of a novel monoclonal antibody (anti-LHK mAb) that has apoptosis-inducing activity against colon cancer Colo205 cells. METHODS: The mechanism of anti-LHK mAb-induced cell death was assessed by microscopic morphology, Annexin V/Hoechst 33528 staining, and detection of DNA fragmentation. The molecular weight of LHK antigen was determined by Western blotting. Growth inhibition of Colo205 cells induced by anti-LHK mAb was determined by in vitro and in vivo studies. RESULTS: Anti-LHK reacted with a 70-kDa antigen and completely blocked the proliferation of Colo205 cells bearing LHK in vitro in a manner characteristic of apoptosis. Strikingly, anti-LHK mAb suppressed tumor growth in a murine peritoneal dissemination model. CONCLUSIONS: LHK antigen, which is restricted to epithelial cells, may be a novel death receptor that plays a critical role in controlling the growth, invasion, and metastasis of human colon cancer cells. PMID- 16261432 TI - Identification and characterization of novel gut-associated lymphoid tissues in rat small intestine. AB - BACKGROUND: The crypt lamina propria of the mouse small intestine has been shown to harbor multiple tiny clusters filled with c-kit- and interleukin 7 receptor (IL-7R)-positive lympho-hemopoietic cells (cryptopatches; CPs). However, it has remained an open question whether similar lymphoid tissue are present in the gastrointesitinal tract in other animals. In the present study, we investigated whether the small intestine of rats harbored lymphoid tissues similar to mouse CPs. METHODS: Immunohistochemical and flow cytometric analyses were carried out using various antibodies, including those to c-kit and IL-7R molecules. RESULTS: Lymphocyte-filled villi (LFVs), populated predominantly with c-kit- and IL-7 receptor (IL-7R)-positive cells and less with T cell receptor (TCR)-alphabeta T cells were found throughout the small intestine of young adult rats. Although LFVs were absent from fetal rat intestine, they were first detected at around 2 weeks after birth. Notably, in most LFVs that settled in the antimesenteric wall of the small intestine in young adult rats, immunoglobulin M-positive B cells were also detectable at the bottom of the LFVs. In aged rats, lymphocytes in some LFVs displayed a different phenotype, comprising a large B-cell area that included a germinal center. Thus, these clusters represent the first description of isolated lymphoid follicles (ILFs) in the rat small intestine. CONCLUSIONS: The present study provides the first evidence for c-kit- and IL-7R-positive lymphocyte clusters in the rat small intestine. Our data also indicating that LFVs and ILFs may constitute novel organized gut-associated lymphoid tissues in lamina propria of the rat small intestine. PMID- 16261433 TI - Emergency balloon-occluded retrograde transvenous obliteration for gastric varices. AB - BACKGROUND: We evaluated the efficacy of emergency balloon-occluded retrograde transvenous obliteration (B-RTO) performed within 24 h after initial hemostasis for the prevention of rebleeding from ruptured gastric varices. METHODS: From December 1995 to March 2003, 11 patients who had undergone B-RTO within 24 h after the control of gastric variceal bleeding at Maebashi Red Cross Hospital were investigated. They were followed up for complete eradication, recurrence of varices, and rebleeding. Efficacy was determined by endoscopic examination and computed tomography. RESULTS: The 4 patients with acute bleeding from ruptured gastric varices were treated with endoscopic therapy-endoscopic variceal ligation (EVL) in 2 patients, and clipping treatment in 2. Initial hemostasis was achieved in all 4; the other 7 patients had already stopped bleeding at endoscopy. After hemostasis was achieved, emergency B-RTO was immediately performed within 24 h and was successful in all 11 patients. Ten (90.9%) of the 11 gastric varices were obliterated and the other 1 (9.1%) was diminished in size. During the mean follow up period of 1136 days, no rebleeding or recurrence as found. Four patients died during the follow-up period, but none died from variceal bleeding. Survival rates were 90.9% and 70.7%, respectively, at 1 year and 3 years. In 6 patients, development of esophageal varices appeared during the follow-up period, all of which were controlled by usual endoscopic therapy. No severe side effects were found after the B-RTO treatment. CONCLUSIONS: Emergency B-RTO is an effective treatment for the prevention of rebleeding from ruptured gastric varices. PMID- 16261434 TI - Prognostic effects of causative virus in hepatocellular carcinoma according to the Japan integrated staging (JIS) score. AB - BACKGROUND: The Japan integrated staging (JIS) score is recognized to be useful in managing hepatocellular carcinoma (HCC). We evaluated the effects of the causative virus in patients stratified by this system. METHODS: We compared clinicopathologic features, cumulative and tumor-free survival rates, and causes of death between 301 hepatitis C virus-positive patients (HCV group) and 60 hepatitis B virus-positive patients (HBV group). RESULTS: Among patients with low JIS scores (0 or 1), the proportions of patients with high aspartate and alanine aminotranferase activities, moderate-to-severe active hepatitis, and with cirrhosis were significantly higher in the HCV than in the HBV group. Among patients with high JIS scores (2 to 4), the proportion with moderate-to-severe active hepatitis was also significantly higher in the HCV group. In patients with low JIS scores, those in the HCV group had significantly lower tumor-free and cumulative survival rates than those in the HBV group. Although no patient in the HBV group died of causes other than liver disease (HCC or hepatic failure), some patients in the HCV group died of causes other than liver disease. The proportion of patients who died because of HCC recurrence tended to be higher among patients with high JIS scores than among patients with a low JIS score. CONCLUSIONS: The effects of viral status on survival outcomes are greatest in patients with JIS scores of 0 or 1. PMID- 16261435 TI - Hyperbaric oxygen-induced changes in bacterial translocation and acinar ultrastructure in rat acute necrotizing pancreatitis. AB - BACKGROUND: We aimed to investigate the effects of hyperbaric oxygen therapy on bacterial translocation and acinar cell ultrastructure in a rat model of acute necrotizing pancreatitis. METHODS: Forty-eight male Sprague-Dawley rats were randomly divided into three groups. Acute pancreatitis was induced in groups II and III. Groups I and II did not receive any treatment, and group III was treated with hyperbaric oxygen. All surviving animals were killed 48 h after the induction of pancreatitis. Bacterial translocation and histological and ultrastructural changes were determined. RESULTS: The incidence of bacterial translocation in group III was significantly lower in comparison with group II (P<0.001). Histopathological and ultrastructural injury scores were also significantly lower in group III (P<0.001 and P<0.04, respectively). CONCLUSIONS: Hyperbaric oxygen therapy displayed beneficial effects on pancreatic superinfection and or histopathological and ultrastructural changes in experimental necrotizing pancreatitis. PMID- 16261436 TI - Cytomegalovirus cholangitis and pancreatitis in an immunocompetent patient. AB - Cholangitis and pancreatitis associated with cytomegalovirus (CMV) infection in an immunocompetent patient is reported. Endoscopic retrograde cholangiography performed on a 55-year-old man for evaluation of the cause of jaundice and liver dysfunction revealed a distal focal irregular narrowing of the common bile duct. Microscopic findings of the resected specimen showed chronic cholangitis and CMV pancreatitis. Immunohistochemistry disclosed that epithelial cells in the inflamed bile duct were positive for CMV antigen, which was compatible with CMV cholangitis. Inflammation of the biliary tract or pancreas by CMV has been commonly reported as a complication in immunocompromised patients. Our report appears to be a rare case, but suggests that CMV cholangitis or pancreatitis should be considered in the differential diagnoses of common bile duct stenosis or pancreatitis even in immunocompetent individuals. PMID- 16261437 TI - Primary undifferentiated spindle-cell carcinoma of the gallbladder presenting as a liver tumor. AB - Undifferentiated spindle-cell carcinoma (SpCC) of the gallbladder is extremely rare. There is very little information available regarding the characteristics and treatment of this disease. We herein report the unique case of a 76-year-old female patient with a primary SpCC of the gallbladder that presented as a liver tumor. Preoperative radiologic examinations showed a 5-cm liver tumor around the gallbladder bed, and irregular thickening of the gallbladder wall. The patient underwent en-bloc resection of the gallbladder and segments 4b and 5 of the liver (including the liver tumor). Microscopic findings revealed that both lesions consisted mainly of a sarcomatous spindle-shaped component. Small foci of well differentiated adenocarcinoma cells were identified in the gallbladder mucosa. There was a gradual transition between the two different components, thereby implying that these two cell types had a common origin. Immunohistochemical studies showed that the spindle-shaped cells were epithelial in nature. The patient's postoperative course was uneventful. However, she died of recurrent liver disease 6 months after the surgery. In conclusion, we surmised that the sarcomatous spindle cells originated from a carcinomatous component in the gallbladder mucosa through dedifferentiation. Further studies are needed to better understand the characteristics of this deadly tumor, and to establish an effective therapy for it. PMID- 16261439 TI - Cancer in the anal canal, and in an anal fistula, that developed during a longstanding course of Crohn's disease. PMID- 16261438 TI - Gastric myoelectrical activity and serum albumin level in Japanese patients on hemodialysis. PMID- 16261440 TI - Budd-Chiari syndrome after in vitro fertilization in a patient with latent thrombophilia. PMID- 16261441 TI - Budd-Chiari syndrome with underlying homozygous factor V Leiden and heterozygous methylenetetrahydrofolate reductase mutations. PMID- 16261442 TI - Metastasis of hepatic angiosarcoma to the stomach. PMID- 16261444 TI - Metallic stent placement or gastroenterostomy for gastric outlet obstruction caused by gastric cancer? PMID- 16261443 TI - 18F-FDG PET for hepatocellular carcinoma presenting with portal vein tumor thrombus. PMID- 16261445 TI - Current status of monoclonal antibodies as therapeutic agents for solid cancer. PMID- 16261446 TI - BMP-2 prevents apoptosis of the N1511 chondrocytic cell line through PI3K/Akt mediated NF-kappaB activation. AB - The signal transduction pathway by which bone morphogenetic protein-2 (BMP-2) regulates apoptosis in chondrocytes remains largely unknown. We investigated the involvement of phosphatidylinositol 3-kinase (PI3K)/Akt-mediated NF-kappaB activation by BMP-2 stimulation in the modulation of this antiapoptotic process in a chondrocytic cell line, N1511. BMP-2 prevented apoptosis through the inhibition of caspase-3 and -9 and an increase in Bcl-xL expression, and this antiapoptotic effect was inhibited by Noggin. Not only was NF-kappaB p65 activated transiently in the early phase (5-15 min) after treatment with BMP-2 but p65 at serine 536 was phosphorylated from 5 min as well. Akt was rapidly phosphorylated in response to BMP-2 treatment; however, the inhibition of PI3K by Wortmannin markedly reduced the phosphorylation of Akt by BMP-2. Wortmannin also decreased the NF-kappaB transcriptional activity that was up-regulated by BMP-2. Thus, BMP-2-induced NF-kappaB activation is mediated by PI3K/Akt signaling. Wortmannin treatment inhibited the antiapoptotic effect of BMP-2. These data indicate that BMP-2 can utilize a new signal transduction pathway in the NF kappaB activation system, which plays a crucial role in the survival of the N1511 chondrocytic cell line. PMID- 16261447 TI - The effects of heat on the biological activity of recombinant human bone morphogenetic protein-2. AB - This study was designed to investigate effects of heat on the bone-inducing activity of recombinant human bone morphogenetic protein (rhBMP)-2. rhBMP-2 samples were heated at 50, 70, 90, or 100 degrees C for 15 min, or 1, 2, 4, or 8 h, or autoclaved at 120 degrees C for 15 min. The bone-inducing activity of the rhBMP-2 before and after heating was assayed in in vivo and in vitro systems. For the in vivo assay, 5 microg rhBMP-2 samples were impregnated into porous collagen disks (6 mm in diameter, 1 mm thickness), freeze dried, and implanted into the back muscles of ddY mice. Three weeks later, the implant was harvested from the host and examined for ectopic new bone tissue by radiography. The new bone mass was quantified by single-energy X-ray absorptiometry. The in vitro activity of the rhBMP-2 was assayed by adding the BMP sample at a concentration of 100 ng/ml to cultures of MC3T3-E1 cells. After 48 h, the alkaline phosphatase activity was measured. After heating at 50 degrees or 70 degrees C, no significant reduction in bone-inducing activity was noted in either in vivo or in vitro assay systems unless the protein was exposed to sustained heat at 70 degrees C for 8 h, based on in vitro assay data. However, heating above 90 degrees C and for longer periods led to a decrease in the biological activity of the rhBMP-2 in a time- and temperature-dependent manner. rhBMP-2 was rendered inactive when exposed to temperatures at or in excess of 120 degrees C. PMID- 16261448 TI - Expression profiles of BMP-related molecules induced by BMP-2 or -4 in muscle derived primary culture cells. AB - The formation of ectopic bone in muscle following the implantation of decalcified bone matrix led to the search and eventual discovery of bone morphogenetic proteins (BMPs) in bone matrix. The precise sequence of molecular events that underpin the cellular transformation of undifferentiated mesenchymal cells into bone has not been established, and is the subject of this study. Northern and Western blot analyses were used to examine changes in gene expression of cells treated with BMP-2 or -4. The molecules, which included BMP receptors (BMPRs), Noggin (a BMP-specific antagonist), osteocalcin (OC), Smad-4, and MyoD, were examined at messenger RNA (mRNA) and protein levels. The changes in expression of these molecules were followed in mouse muscle-derived primary culture cells, and osteoblastic or nonosteoblastic embryonic cell lines. We show the early up regulation of BMPR-1A, -2, Noggin, OC, and Smad-4 in muscle-derived primary culture cells in a dose-dependent manner in response to BMP-2 or -4. MyoD expression was not detected after BMP stimulation. The differential expression of these positive and negative regulators of BMP signaling points to a potential regulatory mechanism for bone induction in mesenchymal cells. PMID- 16261449 TI - Comparison of two assays for fibroblast growth factor (FGF)-23. AB - FGF-23 was recently shown to be involved in the development of several hypophosphatemic diseases, including X-linked hypophosphatemic rickets/osteomalacia (XLH) and tumor-induced rickets/osteomalacia (TIO). FGF-23 is processed between Arg179 and Ser180, and only full-length FGF-23 was shown to cause hypophosphatemia. Two assays for FGF-23 have been reported. One assay detects only full-length FGF-23. In contrast, the C-terminal assay recognizes both full-length and processed C-terminal fragment of FGF-23. However, discrepant results concerning circulatory levels of FGF-23 in patients with TIO and XLH have been reported using these two assays. We simultaneously measured FGF-23 levels in 13 patients with adult-onset hypophosphatemic osteomalacia and 29 patients with XLH by these two assays. The full-length assay indicated that FGF-23 was above the upper limit of the reference range in all patients with osteomalacia and in 24 of 29 patients with XLH. However, the C-terminal assay in dicated that FGF-23 was within the reference range in 3 of 13 patients with osteomalacia and 16 of 29 patients with XLH. In addition, there was no correlation between FGF-23 levels measured by these assays in patients with XLH whose FGF-23 was within the reference range by C-terminal assay. These results indicate that FGF-23 within the reference range by C-terminal assay does not rule out an increase in full length FGF-23. In addition, because FGF-23 was high in most of these hypophosphatemic patients, these results support the notion that FGF-23 plays a major role in the development of hypophosphatemia in patients with TIO and XLH. PMID- 16261450 TI - Polarization and secretion of cathepsin K precede tartrate-resistant acid phosphatase secretion to the ruffled border area during the activation of matrix resorbing clasts. AB - The activation sequence of clasts (the designation clast was used because ultrastructurally in this tissue, it is not always possible to differentiate between chondroclasts sitting on cartilage and osteoclasts sitting on bone matrix) was studied in vivo using the healing of low-phosphate, vitamin D deficiency rickets as a model system. Thus, the bones of 7-week-old rachitic animals were analyzed with a combination of morphological, biochemical, and molecular biological methods at 48 and 72 h, respectively, after change to normal food. A quantitative ultrastructural analysis showed that the number of clast profiles exhibiting the characteristic polarized features of actively resorbing cells, i.e., ruffled borders and clear zones, had reached normal levels after 48 h. By combining the data with quantitative analyses by the immunogold technique, we demonstrated that cathepsin K secretion was coupled to ruffled border formation in clasts irrespective of whether the number of polarized clasts was low (in rickets) or high (in healing). In contrast, the levels of tartrate resistant acid phosphatase (TRAP) both between ruffles and in the outside matrix adjoining the ruffled border were low in polarized clasts both in rickets and at the early (48 h) healing time-point, but were increased at the latest (72 h) healing time-point. Interestingly, expression of TRAP and the cathepsin K at the mRNA level, as well as protein expression and the activity of TRAP, were not different during the healing sequence. Although the two enzymes are confined to the same clast populations, their secretion during the resorption process is apparently differentially regulated: cathepsin K secretion is coupled to ruffled border formation in clasts, whereas TRAP is secreted at a later stage during the resorption sequence, suggesting a role for secreted TRAP as a modulator of resorptive activity. PMID- 16261451 TI - Low-dose parathyroid hormone and estrogen reverse alkaline phosphatase activity suppressed by dexamethasone in mouse osteoblastic cells. AB - Glucocorticoid (GC)-induced osteoporosis (GIO) is frequently seen in patients with excessive GC. Numerous questions remain to be clarified about the pathogenesis and treatment of GIO, and the mechanism of GC-inhibited bone formation is not well known. Several studies suggest that parathyroid hormone (PTH) and hormone replacement therapy are effective for GIO. We therefore investigated whether PTH and estrogen would affect cell proliferation and alkaline phosphatase (ALP) activity inhibited by dexamethasone (Dex) in mouse osteoblastic cell-line MC3T3-E1 cells. Low-dose (10(-11) M) PTH as well as 10(-8) M 17-beta-estradiol (17beta-E2) significantly attenuated Dex-inhibited ALP activity, although 10(-8) M PTH did not affect it. ICI 182780 (10(-8) M) antagonized the effects of 17beta-E(2) on Dex-suppressed ALP activity. Neutralizing anti-IGF-I antibody (3 microg/ml) blocked the reverse effects of 17beta-E2 on ALP activity suppressed by Dex. PTH (10(-11) M), but not 17beta-E2, significantly attenuated [3H]thymidine incorporation inhibited by Dex. On the other hand, PTH and estrogen did not affect the level of 11-beta-hydrosteroid dehydrogenase type I mRNA increased by Dex. In conclusion, the present study demonstrated that low-dose PTH and estrogen reversed Dex-inhibited ALP activity in the mouse osteoblastic cell-line. PMID- 16261452 TI - Effects of risedronate on femoral bone mineral density and bone strength in sciatic neurectomized young rats. AB - Immobilization induces a rapid loss of bone density and bone strength in rats. The purpose of the present study was to examine the effects of risedronate (Ris) on the femoral bone density and bone strength of sciatic neurectomized young rats. Forty male Sprague-Dawley rats, 6 weeks of age, were randomized by the stratified weight method into the following four treatment groups of 10 rats each: sham-operation, bilateral sciatic neurectomy (NX), NX + low-dose Ris (0.25 mg/kg/day, orally), and NX + high-dose Ris (0.5 mg/kg/day, orally). After 8 weeks of feeding, the volumetric bone mineral density (vBMD) and stress strain index (SSI) of the femoral distal metaphysis and middiaphysis of the rats were measured by peripheral quantitative computed tomography. The mechanical properties of the femoral distal metaphysis and middiaphysis were measured by the compression and three-point bending tests, respectively. The femoral length was also measured. As compared with the findings in the sham-operated controls, NX resulted in a loss of femoral length, cancellous vBMD, SSI, maximum load, stiffness, and breaking energy of the femoral distal metaphysis; there was also loss of cortical thickness, SSI, maximum load, and stiffness of the femoral middiaphysis, with no significant effects on the cortical vBMD or breaking energy of the femoral middiaphysis. High-dose Ris increased the vBMD to values higher than those in the sham-operated controls, and prevented the loss of SSI, maximum load, and stiffness of the femoral distal metaphysis, while low-dose Ris prevented the loss of cancellous vBMD of the femoral distal metaphysis. Neither high- nor low-dose Ris affected any of the cortical bone parameters of the femoral middiaphysis, except for cortical thickness, or the femoral length. These findings suggest that Ris may prevent immobilization-induced loss of cancellous bone density and bone strength in a dose-dependent manner without interfering with bone growth, but has no apparent effects on the cortical bone in sciatic neurectomized young rats. The results of the present preclinical study should be taken into consideration prior to the commencement of Ris treatment for disabled children. PMID- 16261453 TI - Effects of pubertal development, height, weight, and grip strength on the bone mineral density of the lumbar spine and hip in peripubertal Japanese children: Kyoto kids increase density in the skeleton study (Kyoto KIDS study). AB - The effects of growth and pubertal development on the bone mineral density (BMD) of the lumbar spine and hip in peripubertal Japanese children were studied as a basis for evaluating the effects of modifiable factors on bone mass gain. The study comprised bone mass measurements in the lumbar spine (L2-4), femoral neck, and total hip using dual-energy X-ray absorptiometry as well as body size measurements and detailed interviews on medical history and pubertal status. The subjects were 404 first-grade students in three junior high schools (129 boys and 275 girls, mean age 12.8 +/- 0.3 years) with no diseases or medication that would affect bone metabolism. BMD at each site showed an increasing trend with physical growth and sexual maturity. Significant positive correlations were observed between BMD at every skeletal site and height, weight, and grip strength in pre- and postpubertal boys and girls. In multiple regression analyses, pubertal development had a significant positive independent effect on BMD at every skeletal site in girls, but not in boys. Physical and pubertal development showed major effects on BMD, but the magnitude of these effects differed in boys and girls, even if they were of the same age. We conclude that confounding factors due to physical and pubertal development should be taken into consideration in different ways for boys and girls in investigations on the effects of environmental or behavioral factors on bone mass acquisition in peripubertal children. PMID- 16261454 TI - Determinants of peak bone mineral density and bone area in young women. AB - Osteoporosis is a disease caused by compromised bone strength, and individuals with a high peak bone mass at a young age are likely to have a high bone mass in old age. To identify the clinical determinants of peak bone mass in young adult women, 418 southern Chinese women, aged 20-39 years, were studied. Low bone mass was defined as areal bone mineral density (aBMD) Z-score < -1 at either the spine or total hip. Within the cohort, 62 (19.0%) and 86 (26.4%) women had low aBMD at the spine and hip, respectively. Regression model analysis revealed that low body weight (<44 kg) was associated with an 8.3-fold (95% CI, 3.7-18.9) and a 6.8-fold (95% CI, 3.0-15.6) risk of having low aBMD at the spine and hip, respectively. Low body weight was also predictive of low volumetric BMD (vBMD) at the spine (odds ratio (OR) 7.8, 95% CI, 3.1-20.1) and femoral neck (OR 3.0, 95% CI, 1.3 7.1). A body height below 153 cm was associated with a 4.8-fold risk in the small L2-4 bone area (95% CI, 2.3-9.8) and a 3.9-fold risk in the small femoral neck area (95% CI, 1.9-8.1). Delayed puberty (onset of menstruation beyond 14 years) was associated with a 2.2-fold (95% CI, 1.0-4.9) increased risk of having low aBMD at the hip. Physical inactivity was associated with a 2.8-fold risk of low spine vBMD (OR 2.8, 95% CI, 1.1-6.7) and a 3.3-fold risk of low hip aBMD (95% CI, 1.0-10.0). Pregnancy protected against low spine aBMD (OR 0.4, 95% CI, 0.1-1.2) and spine vBMD (OR 0.1, 95% CI, 0.0-1.0), low femoral neck vBMD (OR 0.3, 95% CI, 0.1-1.1) and small L2-4 bone area vBMD (OR 0.3, 95% CI, 0.1-1.1). In conclusion, this study identified a number of modifiable determinants of low peak bone mass in young adult women. Maintaining an ideal body weight, engaging in an active lifestyle, and diagnosing late menarche may enable young women to maximize their peak bone mass and so reduce their risk of osteoporosis in later life. PMID- 16261455 TI - Bone mineral density in girls and boys at different pubertal stages: relation with gonadal steroids, bone formation markers, and growth parameters. AB - Puberty has a key role in bone development. During puberty, several nutritional and hormonal factors play a major role in this process. The aim of this study was to determine the changes in areal bone mineral density (BMD), gonadal steroids, bone formation markers, and growth parameters in healthy Turkish pubertal girls and boys at different pubertal stages. In additional, we aimed to detect the relationship between BMD, sex steroids, and growth parameters, and to reveal the most important determinant of BMD in the pubertal period. BMD of the lumbar spine and total body was performed by dual-energy X-ray absorptiometry (Lunar DPX series) in 174 healthy pubertal children (91 girls, 83 boys), aged 11-15 years. Height and weight were measured. Pubertal stages were assesed. Bone formation markers and gonadal steroids were measured. BMD values significantly increased until stage IV in girls. In boys, BMD values also increased during puberty (P < 0.05), but it was significantly higher in stage IV compared with that in other pubertal stages (P < 0.01). Testosterone levels increased until stage IV in both sexes, particularly in boys. Estrogen levels significantly increased during puberty in girls, whereas it was significantly higher at stage IV in boys (P < 0.001). Bone-specific alkaline phosphatase (BAP) level was higher in early and midpuberty, and decreased in late puberty in girls (P < 0.001). BAP level was higher in stage IV in boys. Osteocalcin level was shown not to change significantly in pubertal stages. There was a modest correlation between BMD values and estrogen and testosterone levels in boys. In girls, there was a correlation between BMD values and estrogen levels only (P < 0.05). Weight was significantly associated with BMD in both sexes (P < 0.05). Estrogen had a significant influence on BMD in boys and girls. In conclusion, bone mass increased throughout puberty in both sexes. Peak bone mass was not achieved in girls, but was obtained at stage IV in boys. Bone formation markers were good predictors of bone mass in girls, but not in boys. Estrogen level made the greatest contribution to bone mineral acquisition in boys and girls. The achievement of peak bone mass was sustained by estrogen in boys. The major independent determinant of BMD in both sexes was weight. PMID- 16261456 TI - Independent effect of endogenous dehydroepiandrosterone-sulphate levels and birth weight on bone turnover parameters in young adults. AB - Data indicate that bone turnover is higher in young adults born with a low birth weight (LBW). Moreover, several data support the presence of altered adrenal hormone production in this population. The aim of our study was to investigate whether there is any connection between altered bone homeostasis and adrenal hormone levels. Bone mineral density (BMD), serum osteocalcin (OC), and urinary deoxypyridinoline (DPD) excretion were related to dehydroepiandrosterone-sulphate (DHEAS), cortisol, estradiol, testosterone, and sex-hormone binding globulin (SHBG) levels in 47 healthy young women (of those, 33 were LBW) and 65 healthy young men (of those, 49 were LBW). The age of the subjects was 19-21 years. BMD values were normal and did not correlate with any of the factors investigated. Cortisol did not have any independent effect on bone turnover parameters in either men or women. In women, birth weight, DHEAS levels, and free estradiol index were responsible for almost 50% (corrected r2= 0.45) of serum OC variability. Independent positive associations were observed between DHEAS and OC, and between DHEAS and DPD excretion. In men, birth weight and DHEAS levels together were responsible for more than one-third (corrected r(2) = 0.36) of the variability of serum OC. In contrast with women, DHEAS and OC were inversely correlated in men. Our results suggest that bone turnover depends on the subjects' birth weight. Moreover, DHEAS is also an independent determinant. The effect of DHEAS on bone turnover is different in women and men. DHEAS increases bone turnover in fertile women, while it decreases this in men. PMID- 16261457 TI - Serum 25-hydroxyvitamin D levels and activities of daily living in noninstitutionalized elderly Japanese requiring care. AB - To date, no study has investigated the nutritional status of vitamin D in frail elderly people living at home. The purposes of this study were to assess serum 25 hydroxyvitamin D (25[OH]D) levels and associated factors in noninstitutionalized elderly people who had various levels of physical disability, and to propose an adequate vitamin D nutritional status for the elderly by interpreting the serum 25(OH)D levels in relation to serum parathyroid hormone (PTH) levels in this population. Health examinations were conducted in the winter and summer of 2003. The subjects were 143 elderly people in the winter, and 120 elderly people in the summer, who all used the long-term care insurance system at home. Serum 25(OH)D concentrations were determined with a chemiluminescence protein-binding assay, and serum intact PTH concentrations were determined with an immunoradiometric assay. The subjects' disease histories and lifestyle information were obtained through an interview. Activities of daily living (ADL) levels were evaluated using the Barthel index, and grip strength was measured with a digital hand dynamometer. Average serum 25(OH)D levels in the winter and summer were 54.2 nmol/l (SD 29.0) and 53.3 nmol/l (SD 32.3), respectively, and intact PTH concentrations in the winter and summer were 4.2 pmol/l (SD 1.8) and 4.3 pmol/l (SD 1.8), respectively. The proportion of people who had a low 25(OH)D (<30 nmol/l) and high intact PTH levels (>6.9 pmol/l) were 15%-20% and 8%, respectively. Significant predictors of low serum 25(OH)D concentrations were low ADL levels, female sex, and low fish consumption in both seasons. Serum 25(OH)D concentrations of less than 50 nmol/l were associated with elevated serum intact PTH concentrations. In conclusion, elderly people requiring care at home are at high risk of hypovitaminosis D, and their low serum 25(OH)D levels are mainly associated with low ADL levels. In addition, maintenance of serum 25(OH)D concentrations above 50 nmol/l may prevent hypovitaminosis D-induced hyperparathyroidism. PMID- 16261458 TI - Urinary excretion of type I collagen cross-linked N-telopeptide and serum bone specific alkaline phosphatase analysis to determine the correlation of age and back-pain related changes in elderly women. AB - We evaluated bone turnover using biochemical markers in 273 women over 60 years of age with suspected osteoporosis. Their age range was 60-96 years, with an average of 72.7 years. Patients with disorders that might affect bone metabolism were excluded. Those complaining of back pain within 6 months before or after examination were assigned to the back-pain group. Serum bone-specific alkaline phosphatase (BAP) was measured as a bone formation marker, and the urinary N terminal telopeptide of type I collagen (NTX) was measured as a bone resorption marker. Both BAP and NTX levels increased in individuals over 60 years of age; moreover, these markers were significantly higher in subjects aged over 80 years. Among elderly patients, both BAP and NTX levels were significantly higher in those with back pain than in those without, and in a similar way were observed to increase in parallel with age from the 60-year point. We found that both NTX and BAP increased with aging, and that the increase in these bone turnover markers was closely related to back pain. We also showed that NTX and BAP levels increased significantly in women over 80 years, and specifically in patients with back pain. PMID- 16261459 TI - The VDR, COL1A1, PTH, and PTHR1 gene polymorphisms are not associated with bone size and height in Chinese nuclear families. AB - We tested the relationship of the ApaI, Eco31I, BstBI, and (AAAG)n polymorphisms in the vitamin D receptor (VDR), collagen type I alpha-1 (COL1A1), parathyroid hormone (PTH), and parathyroid hormone (PTH)/PTH-related peptide receptor (PTHR1) genes with variations in bone size (BS) and height. Population stratification, total-family association, and within-family association were used to test these relationships in 400 Chinese nuclear families with a total of 1256 individuals. The BS at hip and spine was measured using a Hologic QDR 2000 dual-energy X-ray absorptiometry (DXA) scanner. The minor allele frequencies were 29.2%, 36.0%, and 14.0% for the VDR-ApaI, COL1A1-Eco31I, and PTH-BstBI markers, respectively. (AAAG)5 and (AAAG)6 of the PTHR1 gene are two major alleles in the Chinese people. Significant population stratification was found between the spine BS and PTHR1-(AAAG)5 (P = 0.048) and PTHR1-(AAAG)6 (P = 0.023), as well as between PTHR1 (AAAG)5 and height (P = 0.048), but we did not detect any significant within family association or total-family association between the VDR, COL1A1, PTH, and PTHR1 gene polymorphisms and the variations in BS and height in our sample. Our results do not support that the VDR, COL1A1, PTH, and PTHR1 genes have an important influence on the variation in BS and height in our Chinese population. PMID- 16261460 TI - Effects of a low sodium diet on bone metabolism. AB - Osteoporosis is a serious public health problem, and dietary interventions may potentially be helpful in preventing this disorder. The purpose of this study was to determine the effects of a low sodium diet on bone metabolism in postmenopausal women. This was a longitudinal study to determine the effects of a low sodium (2-g/day) diet on bone. Forty postmenopausal African-American and Caucasian women were enrolled in a 2-g/day sodium diet for 6 months. Sodium and calcium excretion, bone turnover, and calcitropic hormones (intact parathyroid hormone (PTH) and 1,25 dihydroxyvitamin D) were measured before and 6 months after the intervention. In women who had baseline sodium excretions equal to or greater than the average sodium intake in the United States (> or =3.4 g/day), the low sodium diet resulted in significant decreases in sodium excretion (P = 0.01), in calcium excretion (P = 0.01), and in a biomarker of bone turnover, aminoterminal propeptide of type I collagen (P = 0.04). However, there were no significant changes in calcitropic hormones, including intact PTH (P = 0.97) or 1,25 dihydroxyvitamin D (P = 0.49) with the low sodium diet. These findings suggest that in postmenopausal women with sodium intakes > or =3.4 g/day, a low sodium diet may have benefits for skeletal health. PMID- 16261461 TI - Hungry bones without hypocalcaemia following parathyroidectomy. AB - Hungry bone syndrome is a complication of parathyroid surgery where the correction of primary hyperparathyroidism is associated with rapid bone remineralization, causing severe and prolonged hypocalcaemia [1]. We present a case of hungry bones where although there was rapid bone formation, the serum calcium was maintained within normal limits owing to the development of secondary hyperparathyroidism. PMID- 16261462 TI - How to decrease pain at rapid injection of propofol: effectiveness of flurbiprofen. AB - PURPOSE: Many studies have been conducted on how to decrease propofol injection pain, but none has been completely successful. In the present study, the most effective method was investigated by adding lidocaine or a nonsteroidal antiinflammatory drug or by changing the solvent. METHODS: A total of 250 patients scheduled for general anesthesia were divided into five groups. Anesthesia was induced with intravenous administration of flurbiprofen 50 mg followed immediately by propofol in a long-chain triglyceride (LCT) 2 mg.kg(-1) (flurbiprofen group, n = 50), flurbiprofen 50 mg followed by propofol LCT 2 mg.kg(-1) 1 min later (flurbiprofen 1 group, n = 50), 2% lidocaine 40 mg followed immediately by propofol LCT 2 mg.kg(-1) (lidocaine group, n = 50), propofol LCT 2 mg.kg(-1) alone (LCT group, n = 50), or propofol in a mixture of medium-chain triglyceride (MCT) and LCT 2 mg.kg(-1) (MCT/LCT group, n = 50). Pain at injection was assessed 10 and 20 s after starting the propofol infusion. RESULTS: The numbers of patients with severe and mild pain were larger in the order: LCT group (10 and 31 patients, respectively) > flurbiprofen 1 group (3 and 19) > or = MCT/LCT group (1 and 14) > or = lidocaine group (2 and 11) > flurbiprofen group (0 and 0). CONCLUSIONS: Flurbiprofen 50 mg i.v. just before propofol injection completely abolished propofol injection pain. When it was administered 1 min before propofol injection it was less effective. PMID- 16261463 TI - Insulin secretion and glucose utilization are impaired under general anesthesia with sevoflurane as well as isoflurane in a concentration-independent manner. AB - PURPOSE: The dose-dependent effects of sevoflurane and isoflurane anesthesia on glucose tolerance were compared in humans. METHODS: A prospective, randomized clinical study was conducted in 30 patients. The 30 patients were divided randomly into three sevoflurane anesthesia groups (0.5, 1.0, and 1.5 minimum alveolar concentration [MAC]) and three isoflurane anesthesia groups (0.5, 1.0, and 1.5 MAC). Induction of anesthesia was accomplished by inhalation of the volatile agent and nitrous oxide. After induction, anesthesia was maintained at the designated MAC for 15 min without surgical stimulation. The intravenous glucose tolerance test (IVGTT) was performed in these 30 patients while they were under general anesthesia and again several days after surgery in 5 of these patients while they were awake, as a control. RESULTS: The insulinogenic index (change in concentration of immunoreactive insulin/change in glucose concentration), the acute insulin response, and rates of glucose disappearance were significantly lower in all anesthesia groups than in the control group. However, the insulinogenic index, acute insulin response, and the glucose disappearance rate did not differ significantly among the six anesthesia groups. CONCLUSION: Sevoflurane anesthesia impairs glucose tolerance to the same degree as does isoflurane anesthesia. Glucose intolerance during sevoflurane or isoflurane anesthesia is independent of agent and dosage up to 1.5 MAC. PMID- 16261465 TI - Propofol attenuates oxidant-induced acute lung injury in an isolated perfused rabbit-lung model. AB - PURPOSE: Reactive oxygen species have been strongly implicated in the pathogenesis of acute lung injury (ALI). Some animal studies suggest that free radical scavengers inhibit the onset of oxidant-induced ALI. Propofol (2,6 diisopropylphenol) is chemically similar to phenol-based free radical scavengers such as the endogenous antioxidant vitamin E. Both in vivo and in vitro studies have suggested that propofol has antioxidant potential. We hypothesized that propofol may attenuate ALI by acting as a free-radical scavenger. METHODS: We investigated the effects of propofol on oxidant-induced ALI induced by purine and xanthine oxidase (XO), in isolated perfused rabbit lung, in two series of experiments. In series 1, we examined the relationship between the severity of ALI and the presence of hydrogen peroxide (H2O2). In series 2, we evaluated the effects of propofol on attenuating ALI and the dose dependence of these effects. The lungs were perfused for 90 min, and we evaluated the effects on the severity of ALI by monitoring the pulmonary capillary filtration coefficient (Kfc), pulmonary arterial pressure (Ppa), and the pulmonary capillary hydrostatic pressure (Ppc). RESULTS: In series 1, treatment with catalase (an H2O2 scavenger) prior to the addition of purine and XO resulted in complete prevention of ALI, suggesting that H2O2 may be involved closely in the pathogenesis of ALI. In series 2, pretreatment with propofol at concentrations in excess of 0.5 mM significantly inhibited the increases in the Kfc values, and that in excess of 0.75 mM significantly inhibited the increase in the Ppa values. CONCLUSION: Propofol attenuates oxidant-induced ALI in an isolated perfused rabbit lung model, probably due to its antioxidant action. PMID- 16261464 TI - Landiolol attenuates the cardiovascular response to tracheal intubation. AB - PURPOSE: The objective of this prospective study was to compare the cardiovascular responses with or without landiolol to the induction of general anesthesia and tracheal intubation. METHODS: Twenty-two patients were randomly allocated to receive a loading dose of landiolol 125 microg kg(-1) min(-1) for 1 min followed by an infusion at 40 microg kg(-1) min(-1) for 4 min, or placebo. Four minutes after landiolol or placebo was started, propofol and succinylcholine were administered. Laryngoscopy and tracheal intubation were performed 1 min after the administration of succinylcholine. Heart rate and blood pressure were measured noninvasively every minute. RESULTS: A significant attenuation of the heart rate and blood pressure response were seen in the landiolol group for 3 min after intubation. Heart rate and systolic blood pressure in the landiolol group were decreased for 2 min before intubation and just before intubation compared with baseline, respectively. CONCLUSION: Continuous administration of landiolol before tracheal intubation results in the attenuation of cardiovascular response for tracheal intubation. PMID- 16261466 TI - Effects of olprinone on hepatosplanchnic circulation and mitochondrial oxidation in a porcine model of endotoxemia. AB - PURPOSE: This study was performed in order to assess the effects of olprinone, a phosphodiesterase III inhibitor, on hepatic oxygen delivery (DO2H), oxygen consumption (VO2H), and mitochondrial oxidation in the liver of a porcine endotoxemia model. METHODS: Fourteen pigs received continuous infusion of endotoxin via the portal vein for 240 min. From t = 150 to t = 240 min, animals were randomly divided into two groups to receive saline (control [CONT]; n = 7), or olprinone (OLP; n = 7) via the central vein. RESULTS: In the OLP group, prior to olprinone treatment at 150 min, endotoxin induced significant decreases in the cardiac index (CI; from 120 +/- 31 to 65 +/- 13 ml.kg(-1).min(-1); P < 0.01) and DO2H (from 3.58 +/- 0.81 to 1.55 +/- 0.49 ml.kg(-1).min(-1); P < 0.01), while VO2H was maintained. After administration of olprinone (from t = 150 to t = 240 min), CI was unchanged, while DO2H increased from 1.55 +/- 0.49 to 1.93 +/- 0.38 ml.kg(-1).min(-1) (P < 0.01) and VO(2)H increased from 0.42 +/- 0.28 to 0.69 +/- 0.38 ml.kg(-1).min(-1) (P < 0.01). At t = 240 min, the oxidation level of cytochrome aa3 was significantly higher in the OLP group than in the CONT group (OLP, 66.2 +/- 19.3% vs CONT, 26.4 +/- 17.3%; P < 0.01). CONCLUSION: Our data for this porcine endotoxemia model suggest that olprinone may have beneficial therapeutic effects in restoring not only systemic and hepatic circulation but also mitochondrial oxidation in the liver. PMID- 16261467 TI - Drugs to facilitate recovery of neuromuscular blockade and muscle strength. AB - Several drugs that quicken recovery from neuromuscular blockade caused by vecuronium in anesthetized patients are reviewed. Ulinastatin, a protease inhibitor, is thought to promote the release of acetylcholine at the neuromuscular junction and increases hepatic blood flow and urine volume. For this reason, ulinastatin quickens recovery from neuromuscular blockade in anesthetized patients receiving vecuronium. Additionally, pretreatment with ulinastatin avoids prolongation of vecuronium-induced neuromuscular blockade in patients with hepatic cirrhosis. Gabexate mesilate is also a protease inhibitor. During a continuous infusion of gabexate mesilate, recovery from neuromuscular blockade was quickened. Amino acid-enriched solution supplies energy to the skeletal muscles and causes an increase in muscle strength. An infusion of amino acid-enriched solution hastens recovery from neuromuscular blockade in anesthetized patients. When amino acids supply energy to the skeletal muscles, they simultaneously produce heat in the skeletal muscles. This thermal generation may be closely related to fast recovery from neuromuscular blockade. Amino acid enriched solution makes recovery from neuromuscular blockade quick and avoids hypothermia during general anesthesia. Milrinone, a phosphodiesterase III inhibitor, is supposed to increase the release of acetylcholine at the neuromuscular junction and make the neuromuscular junction sensitive to acetylcholine. Therefore, recovery from neuromuscular blockade is hastened. Nicorandil enhances membrane K+ conductance in skeletal muscle and increases contraction of the skeletal muscle. Thus, nicorandil quickens recovery from neuromuscular blockade. PMID- 16261468 TI - Anesthetic and airway management of general anesthesia in a patient with Meckel Gruber syndrome. AB - Meckel-Gruber syndrome, characterized by occipital encephalocele, microcephaly, polydactyly, cleft lip or palate, mandibular micrognathism, and anatomical abnormality of the larynx and tongue, along with other associated malformations, is in the list of diseases associated with difficult airway. However, there has been no report on the management of general anesthesia and airway management for such patients. A 2-year-old girl with Meckel-Gruber syndrome was scheduled for cardioplasty and gastrostomy for gastroesophageal reflux under general anesthesia. Preoperative examination revealed obesity, microgenia, dysspondylism, proteinuria, hypoplastic kidneys, and stenosis of the anal canal. Although we anticipated some difficulty with the intubation and prepared several alternative methods for intubation, such as a bronchofiberscope and a laryngeal mask airway, tracheal intubation was completed without difficulty using conventional laryngoscopy after inhalational induction with sevoflurane. Because most patients with this syndrome die before and shortly after delivery, those who survive to some age might have less severe deformities. PMID- 16261469 TI - Combined spinal-propofol anesthesia with noninvasive positive-pressure ventilation. AB - Twenty-three adult patients undergoing repair of inguinal hernia under spinal anesthesia received propofol infusion for sedation with the assist of noninvasive positive-pressure ventilation (NPPV). Circulatory and respiratory parameters, such as percutaneous oxygen saturation, transcutaneous carbon dioxide tension, respiratory rate, tidal volume, blood pressure, and heart rate, were maintained within physiological ranges during the anesthesia. There were no adverse effects. These findings suggest that the application of NPPV in patients receiving propofol infusion for sedation is clinically practicable during anesthesia. PMID- 16261470 TI - Informed consent for anesthesia: survey of current practices in Japan. AB - Anesthesia requires informed consent because it is an invasive procedure with certain risks. However, the state of informed consent for anesthesia in Japan remains unclear. The purpose of this survey was to examine the state of informed consent for anesthesia in Japan. A questionnaire was sent to all hospitals certified by the Japanese Society of Anesthesiologists (n = 854). The questionnaire consisted of four sections: explanation of the anesthesia, method of documentation, consent for anesthesia, and other information such as the hospital's size. A total of 504 (59.0%) questionnaires were completed and returned. At 96.7% of hospitals, an anesthesiologist would explain the scheduled anesthesia. Most departments provide an explanation of dental damage, malignant hyperthermia, and nausea/vomiting. Explanation of anesthesia was standardized at 59.0% of hospitals. A written description was handed out to patients routinely at 61.3% of hospitals. Although consent for anesthesia was obtained at more than 90% of departments, only 59.9% of departments would keep records of having obtained consent. This survey found that the explanation of anesthesia varied among hospitals and was not standardized in Japan. Further attention is needed on how to improve the documentation of informed consent. PMID- 16261471 TI - Postoperative myasthenic crisis successfully treated with immunoadsorption therapy. AB - Myasthenia gravis (MG) is an autoimmune disorder characterized by loss of acetylcholine receptors (AChR) due primarily to the production of anti-AChR autoantibodies. We report a case of MG associated with elevated anti acetylcholine receptor antibody (anti-AChR Ab) and refractory crisis after thymectomy, in which immunoadsorption therapy was used successfully to stabilize myasthenic symptoms and decrease the anti-AChR Ab titer. A 79-year-old woman underwent extended thymectomy under the diagnosis of MG. One day after surgery she suddenly underwent a myasthenic crisis and was successfully resuscitated. Immunoadsorption therapy with a tryptophan-linked polyvinyl alcohol adsorber was performed three times for the purpose of decreasing the anti-AChR antibodies. The anti-AChR Ab titer was reduced by immunoadsorption during each therapy session; however, the level of anti-AChR Ab before immunoadsorption was higher than that of the previous treatment. This case suggests that the absolute serum level of anti-AChR Ab does not always correlate with the severity of the disease. Removal of pathogenic factors, not only anti-AChR Ab but other antibodies with specificities to skeletal muscles or certain components of the complement system, may contribute to effective treatment of myasthenic crisis. PMID- 16261472 TI - Immediate postoperative refeeding in orthopedic surgery is safe. AB - The purpose of this retrospective study was to investigate the morbidity of immediate postoperative refeeding after orthopedic surgery. We included all the 1077 patients who underwent orthopedic surgery between January and December 2003 at our military teaching hospital. General anesthesia was performed in 37% of the patients (n = 398), 24% (n = 259) had combined general and regional anesthesia, and 39% (n = 420) had isolated regional anesthesia (spinal anesthesia and/or peripheral regional anesthesia). After surgery, each patient was allowed free access to solid and liquid food immediately after discharge from the postanesthetic care unit. Although no systematic nausea and vomiting prophylaxis was performed, only 7% (n = 75) of the patients had postoperative nausea and vomiting during the first 48 h. Moreover, neither deglutition trouble nor aspiration syndrome was observed during that period. Our results suggest that immediate postoperative refeeding after orthopedic surgery is safe, does not increase postoperative nausea and vomiting, and probably increases the comfort of patients. PMID- 16261473 TI - Airway obstruction involving a laryngeal mask airway during arthroscopic shoulder surgery. AB - Several earlier reports have described life-threatening airway obstruction during arthroscopic shoulder surgery performed under regional anesthesia, caused by the leakage of irrigation fluid out of the shoulder joint space into the surrounding soft tissues and then the neck and the pharynx. Here, we present a case of airway obstruction that occurred in a patient under general anesthesia. A 77-year-old woman with a rotator cuff rupture who was to undergo right-shoulder arthroscopic surgery was anesthetized with fentanyl and propofol. Her airway was secured with a flexible laryngeal mask airway (LMA). During surgery, the compliance of her breathing bag became gradually poorer, and finally we were not able to ventilate her at an airway pressure of 60 cmH2O. We found that her chest wall, neck, and face were swollen and tense. Laryngoscopy revealed massive swelling of the pharyngeal soft tissues. The vocal cords were not visible. Her trachea was intubated blindly, and adequate ventilation was re-established. She was placed in the Fowler position and furosemide was given intravenously. Her neck and chest swelling were reduced over the next 2 h and she was extubated without any problem. We recommend that physicians should periodically examine the neck of any patient undergoing arthroscopic shoulder surgery, especially when general anesthesia is used, because anesthetized patients cannot complain of breathing difficulty and the airway swelling may progress until it becomes life threatening. PMID- 16261475 TI - Methemoglobinemia induced by automobile exhaust fumes. AB - Although methemoglobinemia is an uncommon disorder, it should always be considered in the differential diagnosis of cyanosis. Major causes of acquired methemoglobinemia are nitrates, aniline, and analgesics, though rare cases have been reported to have been caused by automobile exhaust fumes. A 24-year-old man had inhaled a large amount of automobile exhaust fumes, intending to commit suicide. He had become unconscious, with dilated pupils and symptoms of cyanosis. Arterial hemoglobin oxygen saturation (Sp(O2)) was 86%, with a methemoglobin level of 44.3% and a carboxyhemoglobin level of 0%, while electrolytes, blood urea nitrogen, creatine, and glucose measurement results were normal. He was treated with methylene blue 250 mg (approximately 4 mg/kg) through a nasogastric tube. Four hours after the treatment, because the methemoglobin level was slightly above normal (2.2%), we added 180 mg of methylene blue. The results of final arterial blood gas analysis were a methemoglobin level of 0.4% and a carboxyhemoglobin level of 0.8%. He recovered uneventfully and returned home by himself the next day. To summarize, we successfully treated, with methylene blue given through a nasogastric tube, a young man who had developed severe methemoglobinemia from inhaling automobile exhaust fumes. PMID- 16261474 TI - Combined neurolytic block of celiac, inferior mesenteric, and superior hypogastric plexuses for incapacitating abdominal and/or pelvic cancer pain. AB - Thirty-five patients with extensive abdominal or pelvic cancer who suffered uncontrolled, diffuse, extensive, and incapacitating pain were treated with a combination of neurolytic celiac plexus block (CPB), inferior mesenteric plexus block (IMPB), and superior hypogastric plexus block (SHGPB). The combination of neurolytic CPB, IMPB, and SHGPB was performed with alcohol, mainly using a transintervetebral disc approach. The combination neurolysis produced effective immediate pain relief in all the patients (visual analog scale (VAS), reduced from 8.8 +/- 0.2 to 0). This pain relief persisted during the first 3 months (VAS, 2.3 +/- 0.5) or until death. Morphine consumption was significantly decreased for the first 1 month (from 96 +/- 29 mg to 31 +/- 10 mg per day) after the neurolysis and thereafter continued to be lower than before the surgery, though not significantly so. No serious complications were observed to have been caused by the neurolytic procedure on the three sympathetic plexuses. Our preliminary clinical results suggest that the combination of neurolytic CPB, IMPB, and SHGPB improves the quality of life of patients who have incapacitating cancer pain, by reducing both the intensity of the pain and their opioid consumption, without serious complications. This combination procedure may provide a new therapeutic option for pain relief in patients with advanced cancer. PMID- 16261477 TI - Abnormally low bispectral index and isoelectric electroencephalogram observed after administration of small doses of propofol during induction of anesthesia. AB - We describe a case in which an unexpectedly, abnormally low bispectral index value (BIS = 4) and an almost isoelectric electroencephalogram (EEG) pattern were observed during typical induction of anesthesia with propofol. Starting 2 min after the beginning of propofol administration (1.26 mg kg(-1)), the EEG recordings showed burst and suppression pattern for the next 12 min. The EEG during this period was characterized by gradual prolongation of suppression periods until the appearance of the isoelectric line. After that, burst activity returned and eventually the burst suppression pattern disappeared. We excluded the possibility of ischemic brain damage, and the evidence increasingly points toward a greater sensitivity to propofol. The findings described in this case report support the thesis that there is a wide variability in the responses of patients to propofol that cannot be detected without continuous monitoring of cortical electrical activity. PMID- 16261476 TI - Target-controlled infusion of propofol for a patient with myotonic dystrophy. AB - We present a patient with myotonic dystrophy (MD) who was anesthetized with propofol using a target-controlled technique for electrophysiologic examination and cardiac catheter ablation. The patient became apneic unexpectedly at the same time when he fell asleep, with effect-site propofol concentration of 1.6 microg ml(-1). We had to insert a laryngeal mask airway (LMA), and mechanical ventilation was performed. The patient opened his eyes on verbal command at an effect-site concentration of 1.2 microg ml(-1) after the procedure. This concentration (1.2 microg ml(-1)) was slightly lower than our institutional average for adult male patients (1.5 +/- 0.2 microg ml(-1)). However, the time from the end of anesthesia to the patient's awakening was about 10 min. We considered that emergence from anesthesia was not delayed in this case. Careful titration of propofol by target-controlled infusion (TCI) enabled to evaluate the patient's sensitivity to propofol. We conclude that TCI of propofol was a useful anesthetic technique in the MD patient. Respiratory depression might occur in MD patients at low propofol concentrations. Precise control and titration over target propofol concentration is important in anesthetic management for MD patients. PMID- 16261478 TI - Volatile anesthetics constrict pulmonary artery in rabbit lung perfusion model. AB - Volatile anesthetics are generally considered to possess a vasodilator action. Some of their actions on pulmonary vessels, however, are not clearly understood. We examined the effects of various volatile anesthetics on pulmonary vessels using an in situ rabbit isolated-lung perfusion model. We prepared a rabbit constant-flow lung-perfusion model by sending blood to the pulmonary artery and removing blood from the left atrium, and observed the changes in pulmonary arterial perfusion pressure caused by inhalation of 0.5, 1, 2, and 3 minimum alveolar concentration (MAC) volatile anesthetics: halothane, enflurane, isoflurane, and sevoflurane, in random order. These volatile anesthetics increased pulmonary arterial perfusion pressure in a dose-dependent manner and caused the pulmonary arteries to constrict. In particular, halothane at all concentrations induced significantly greater pulmonary vasoconstriction than the other volatile anesthetics. Therefore, it is suggested that volatile inhalation anesthetics induce the pulmonary arteries to constrict, and halothane exhibits the most potent pulmonary vasoconstrictor effect among the volatile anesthetics tested. PMID- 16261479 TI - Postoperative necrosis of the anterior part of the cervical trachea following thyroidectomy. PMID- 16261480 TI - Shaw scalpel for breast mastectomy in a pacemaker-implanted patient. PMID- 16261481 TI - Using digital media to keep anesthesia history alive. AB - Multimedia applications have been developed to explain many aspects of anesthesiology, including anesthesia machines, cardiopulmonary bypass systems, airway management, anesthetic pharmacology, regional anesthesia, and more recently, the history of the specialty. This article describes how computer assisted instruction and multimedia have made it possible for an anesthesia department in a teaching hospital to become involved in the preservation, publication, and distribution of historically related anesthesia materials. The production of a digital video disc (DVD) based on the career of Leroy D. Vandam is discussed to illustrate the value of digital media in preserving, rejuvenating, and distributing anesthesia-related historical materials. PMID- 16261482 TI - Portfolio assessment. PMID- 16261483 TI - Mentoring veterinary students. PMID- 16261484 TI - Why self-directed learning is not learned and practiced in veterinary education. AB - Veterinary education as it is frequently implemented creates numerous impediments for the majority of students to achieve self-directed learning. Self-directed learning is a state in which the student is prepared to engage in learning activities defined by him- or herself. Essential components of veterinary education (i.e., curriculum and assessment) inhibit the development of self directed learning in many ways. This article will make suggestions on how we can promote self-directed learning within traditional veterinary education and discuss how teaching based upon the principles of problem-based learning (PBL) may foster self-directed learning. PMID- 16261486 TI - The veterinarian scientist: an endangered species. PMID- 16261487 TI - Veterinarians in biomedical research: a perilous future? PMID- 16261488 TI - Musings of a Connecticut Yankee in King Arthur's court: antemortem analysis of the veterinary profession. PMID- 16261489 TI - Funding opportunities for research and graduate education: how the NIH can help you. PMID- 16261490 TI - Funding opportunities for veterinary medicine with USDA competitive programs. AB - Dramatic changes are underway in how public higher education and professional training are funded. The shift from a public to a beneficiary funding model is driving increases in tuition for students, the collection of user fees, and fee for-service activities and has focused increased attention on the need for funding support from federal grants and contracts. The US Department of Agriculture (USDA) was the first federal agency to support university-based research and education. While the funding provided by the USDA is now only a fraction of what is available through other federal sources, the USDA remains an important source of funding dedicated specifically to farm-animal-related research and the training of the next generation of veterinary scientists and educators. PMID- 16261491 TI - NSF 101: fundamentals of the proposal review process and funding opportunities at the National Science Foundation. PMID- 16261492 TI - Quality assessment of graduate programs. PMID- 16261493 TI - R(X) for recruitment and retention of veterinarian scientists: money, marketing, mentoring. AB - Veterinary medicine is failing both to sustain its academic base and to meet national needs for research in the fields of comparative medicine (translational research), public health, and food production. The basis for the shortage of veterinarians with research expertise is multi-factorial and related to the substantial commitment of time and money required to obtain both a DVM and advanced training, as well as the lack of motivation among veterinary students to engage in biomedical science. Effective strategies for increasing the number of veterinarian scientists must address these issues using a balanced combination of money, marketing, and mentoring. Success will require not only that we increase and improve opportunities for research training, but also that we create and sustain veterinary college environments that attract, foster, and reward dedication to research. The ''research pipeline'' needs to be transformed into a ''research manifold'' with multiple portals for entry and re-entry of trainees. Age-appropriate educational and mentoring programs should be implemented at K-14, baccalaureate, veterinary college, post-graduate, and junior faculty levels to promote recruitment, training, and retention of veterinarian scientists. New initiatives are especially needed to attract students with primary interests in science and biomedical research to the veterinary profession and to facilitate transition of motivated veterinary graduates from private practice to research careers. Specific examples of such programs are presented and future directions are discussed. PMID- 16261494 TI - VMSTP Combined Degree Training (VMD/PhD): key features of the program at the University of Pennsylvania. AB - The Veterinary Medical Scientist Training Program (VMSTP), a combined degree program at the University of Pennsylvania School of Veterinary Medicine (Penn), has been in existence for approximately 35 years and has an excellent track record of producing veterinary physician-scientists. There are a number of key features of the program that I believe have been crucial to its success. Many of these features relate to how the PhD training component of the combined degree program is accomplished. Rather that describing the veterinary training component of the program, I will describe the PhD training component and how this training intersects with the veterinary curriculum. The key features of the VMSTP program at Penn are (1) admitting the right candidates, (2) placing the PhD training component of the program in the hands of individual graduate groups, (3) being committed to PhD training of veterinary students that is not compromised in terms of quality or time, (4) devising mechanisms to interdigitate VMD and PhD training to generate synergy between the programs, (5) providing continual advice to students from numerous perspectives, and (6) providing the monetary and emotional support needed for the long-term commitment students must make in order to complete the program. PMID- 16261495 TI - Developing scientist leaders for tumultuous times. AB - Leadership is a quality that can be learned. It is a behavior that one practices, and, after lots of practice, it becomes a habit. This is a lesson I learned from my father, who was a career pilot in the US Air Force and instilled this into me and my siblings from a very early age. It is also something I have learned in observing others. I have frequently asked why some people from certain disciplinary backgrounds seem to have an advantage in the leadership area. Think of the backgrounds of our Presidents, for example; so many of them have been attorneys. Members of Congress, as well, also frequently come from that disciplinary background. Key decision makers in government frequently come from economics backgrounds. I have also asked why this is the case. Frequently, the answer seems to be that these disciplines define themselves as being those that create leaders, not that they limit their members' aspirations. Why are so few veterinarians in leadership positions? It seems quite a paradox that they are not. The assets of an education in veterinary medicine are many. The education provides a very broad background in systems biology, medicine, and public health. There are many career paths for veterinarians. Most choose private practice, but, beyond that, career paths exist in industry, particularly the biomedical industry; in trade associations; in government and industry research; and in public health and regulatory positions. There are also many opportunities in academia, certainly in colleges of veterinary medicine but, beyond that, also in human medicine and in the biology disciplines. International opportunities also exist in governmental and non-governmental organizations, such as the Food and Agriculture Organization at the United Nations and the World Health Organization, and in advocacy and lobbying. Veterinarians are also making news these days. The emerging zoonotic diseases that have seized headlines in papers around the world give prominence to veterinarians and the skills they bring to bear in fighting current outbreaks and preventing future outbreaks of these diseases, such as SARS, Ebola, West Nile virus, and even HIV/AIDS. PMID- 16261496 TI - Providing stewardship and leadership for the future of veterinary medical research: a responsibility of the veterinary colleges. AB - As we contemplate responsibilities as well as opportunities in research, it is fair to begin by considering why research is, or should be, important to veterinary schools and colleges, to our profession, and to society. Veterinary research is conducted in many venues, such as colleges of veterinary medicine, veterinary science departments, comparative medicine departments, medical schools, and many other university departments, as well as in industry laboratories, governmental agencies, and other organizations. But schools and colleges of veterinary medicine have a unique and historic responsibility to ensure that our veterinary medical research programs are sufficient in quality, capacity, and depth to meet the research needs of our society. Consequently, my comments will be oriented toward veterinary schools and colleges. PMID- 16261497 TI - Teaching and learning at the Faculty of Veterinary Science, University of Sydney. PMID- 16261498 TI - The Faculty of Veterinary Science at the University of Queensland. PMID- 16261499 TI - The Massey University Veterinary School. PMID- 16261500 TI - Recruiting faculty with clinical responsibilities: factors that influence a decision to accept an academic position. AB - The current opportunities for veterinary clinical specialists in private practice and industry have made recruiting and retaining faculty a major focus for most clinical academic departments. To gain a better understanding of the importance of the various factors considered in accepting an academic position, an electronic survey was distributed to newly hired veterinary faculty with clinical responsibilities. The results suggest that the perceived climate and collegiality within the prospective hiring department is the most important factor influencing the decision to accept an academic position. Salary is the second most important factor. Institutional support for the newly hired faculty member and the reputation and quality of the prospective institution rank as more important than the perceived quality of the local community and the geographic location of the institution. The search process and administrative support are the least important factors. There were no differences between the responses of faculty hired into tenure-track positions and those of faculty hired into clinical-track positions. Focusing on the advantages of a collegial environment, enhancing compensation packages, and using creative and flexible appointments may improve faculty recruitment and retention in clinical academic departments. PMID- 16261501 TI - Experiences and difficulties encountered during a course on veterinary public health with students of different nationalities. AB - Veterinary public health (VPH) issues have received increased attention over the last few years as a result of the rising threat of emerging zoonoses (i.e., those due to globalized trade in animal and animal products and to changes in livestock production systems and the environment). The international dimension of VPH is gradually becoming recognized, and there is a growing need for veterinarians with experience in this field. In order to familiarize (future) veterinarians with the international dimension of VPH, the Department of Public Health and Food Safety of the Faculty of Veterinary Medicine, Utrecht University, has been organizing a course in Veterinary Public Health and Animal Production for over the last 10 years. This course has been intended for Dutch as well as foreign final-year veterinary students and recent veterinary graduates. By bringing together participants from different countries, the course reinforces the international dimension of the issues addressed through the exchange of experiences by the participants themselves. The present article provides information about this course on Veterinary Public Health (VPH): it discusses logistics, didactical approaches, the course program, and the use of information and communication technology (ICT). Special attention is given to the intercultural aspects of higher education, all of which play an important role in the efficient exchange of knowledge between lecturers and students. International courses are an important tool to enable participants to interact in a multicultural environment and address issues that demand international cooperation and a global public health focus. PMID- 16261502 TI - The North Carolina State University College of Veterinary Medicine Turtle Rescue Team: a model for a successful wild-reptile clinic. AB - The North Carolina State University College of Veterinary Medicine (NCSU-CVM) Turtle Rescue Team (TRT) is a veterinary student-run organization that treats native, sick and injured, wild chelonians. First-, second-, and third-year students are responsible for case management, consultation coordination, diagnostic testing within the hospital, and placing of recuperating animals with local wildlife rehabilitators. Several clinical research publications have resulted from the opportunity to work with these wild reptiles. Active student participants can also gain a course credit by attending eight hours of lecture/ seminar related to reptile medicine. With regards to outcome assessment, 86% of survey respondents found the program to be valuable or extremely valuable to their veterinary medical education. The logistics of organizing, supporting, and running this service are discussed, and its value as a clinical learning tool is supported by the results of a survey. PMID- 16261503 TI - Determination of the hyperfine coupling constant and zero-field splitting in the ESR spectrum of Mn(2+) in calcite. AB - The hyperfine coupling constant (A in units of energy or A' in units of magnetic field) and zero-field splitting (D in units of energy or D' in units of magnetic field) in the ESR spectrum of Mn(2+) in calcite were determined. The spreading of the non-central allowed transitions |3/2, m --> |5/2, m, |1/2, m --> |3/2, m, | 3/2, m --> |-1/2, m and |-5/2, m --> |-3/2, m was analysed and the experimental transitions were attributed. Particular relevance was given to the difference between the two types of resonances, shoulder or divergence, and to their origin (M, m). The analysis explains the presence of a weak shoulder between each of the five central doublets |-1/2, m --> |1/2, m. Six independent methods for calculating the hyperfine coupling constant and five methods for calculating the zero-field splitting, based on the analysis of the allowed and forbidden transitions, were provided. The values of the hyperfine coupling constant range from -93.9 to -94.6 G and those of the zero-field splitting range from -79.5 to 80.5 G. A critical evaluation of the advantages and drawbacks of the 11 methods is included: the best value for A' is -94.30 G and that for D' is -79.95 G. PMID- 16261504 TI - Complete assignments of (1)H and (13)C NMR data for two 3beta,8beta epoxymexicanolides from the fruit of a Chinese mangrove Xylocarpus granatum. AB - Three 3beta,8beta-epoxymexicanolides, including xyloccensin K, 6 acetoxycedrodorin and a new one named xyloccensin W, were isolated from the fruit of a Chinese mangrove Xylocarpus granatum. Their structures were determined by spectroscopic analyses. The first complete assignment of (1)H and (13)C NMR data for xyloccensin W was achieved by means of 2D NMR techniques, including (1)H-(1)H COSY, HSQC, HMBC and NOESY spectra. In addition, the confusion of (1)H and (13)C NMR data previously reported for xyloccensin K was clarified. PMID- 16261505 TI - (1)H and (13)C NMR spectral data for a tricyclic derivative of a Diels-Alder adduct. AB - A complete NMR analysis with full assignment for (1)H and (13)C NMR spectral data for 5-(acetyloxy)-3-hydroxy-9,10-dimethoxy-6-oxo-11 oxatricyclo[6.2.1.0(2,7)]undec-2-yl acetate is described. This compound was prepared by rapid hydrogenation of the unstable Diels-Alder adduct obtained from the reaction between 3,4-dimethoxyfuran and 2,5-diacetoxy-1,4-benzoquinone. Full homonuclear hydrogen coupling constants measurements and molecular mechanics calculations were performed for the determination of the relative stereochemistry. PMID- 16261506 TI - (29)Si NMR in LC-NMR analysis of silicones. AB - It is demonstrated using a practical example that indirect detection of (29)Si NMR signals is sufficiently sensitive in LC-NMR stop-flow arrangement to analyze mixtures of siloxane polymers. New cryogenic probes with better signal-to-noise ratio will turn this version of LC-NMR into a routine method for analysis of siloxane polymers. PMID- 16261508 TI - Conformational behavior of temporin A and temporin L in aqueous solution: a computational/experimental study. AB - Molecular dynamics (MD) simulations and circular dichroism (CD) experiments were carried out on aqueous temporin A and L, two short peptides belonging to an interesting class of natural substances known to be active mainly against Gram positive/negative bacteria and fungi. Experimental results indicate the higher propensity of temporin L, with respect to temporin A, in forming alpha-helical structures. These results were revisited by long-timescale MD simulations, in which their alpha-helical propensity was investigated in the absence of trifluoroethanol. Results clearly show the higher stability of alpha-helix conformations in temporin L; moreover, an interestingly strong mechanical analogy emerges since both temporins show the same residue interval (from 7 to 10) as the most energetically accessible for alpha-helix formation. Such studies provide some intriguing structural and mechanical evidence that may help in better understanding and rationalizing the conformational behaviour of temporins in water solution and, ultimately, the inner principles of their microbial targets selectivity and mechanism of action at the level of cell membranes. PMID- 16261509 TI - Conformational spaces of the gastrointestinal antisecretory chiral drug omeprazole: stereochemistry and tautomerism. AB - A study of the conformational spaces of the chiral proton pump inhibitor (PPI) drug omeprazole by semiempirical, ab-initio, and DFT methods is described. In addition to the chiral center at the sulfinyl sulfur atom, the chiral axis at the pyridine ring (due to the hindered rotation of the 4-methoxy substituents) was considered. The results were analyzed in terms of the 5-methoxy and 6-methoxy tautomers and the two pairs of enantiomers (R,P)/(S,M) and (R,M)/(S,P). Five torsion angles were systematically explored: the backbone rotations defined by D1 (N3-C2-S10-O11), D2 (C2-S10-C12-C13), and D3 (S10-C12-C13-N14) and two methoxy rotations defined by D4 (C6-C5-O8-C9) and D5 (C16-C17-O19-C20). Significant energy differences were revealed between the 5- and 6-methoxy tautomers, the extended and folded conformations, and the (S,M) and (S,P) diastereomers. The "extended M" conformation of the 6-methoxy tautomer of (S)-omeprazole was found to be the most stable conformer. PMID- 16261511 TI - Stimulating effect of adaptogens: an overview with particular reference to their efficacy following single dose administration. AB - Plant adaptogens are compounds that increase the ability of an organism to adapt to environmental factors and to avoid damage from such factors. The beneficial effects of multi-dose administration of adaptogens are mainly associated with the hypothalamic-pituitary-adrenal (HPA) axis, a part of the stress-system that is believed to play a primary role in the reactions of the body to repeated stress and adaptation. In contrast, the single dose application of adaptogens is important in situations that require a rapid response to tension or to a stressful situation. In this case, the effects of the adaptogens are associated with another part of the stress-system, namely, the sympatho-adrenal-system (SAS), that provides a rapid response mechanism mainly to control the acute reaction of the organism to a stressor. This review focuses primarily on the SAS mediated stimulating effects of single doses of adaptogens derived from Rhodiola rosea, Schizandra chinensis and Eleutherococcus senticosus. The use of these drugs typically generates no side effects, unlike traditional stimulants that possess addiction, tolerance and abuse potential, produce a negative effect on sleep structure, and cause rebound hypersomnolence or 'come down' effects. Furthermore, single administration of these adaptogens effectively increases mental performance and physical working capacity in humans. R. rosea is the most active of the three plant adaptogens producing, within 30 min of administration, a stimulating effect that continues for at least 4-6 h. The active principles of the three plants that exhibit single dose stimulating effects are glycosides of phenylpropane- and phenylethane-based phenolic compounds such as salidroside, rosavin, syringin and triandrin, the latter being the most active. PMID- 16261512 TI - In vitro GABA-transaminase inhibitory compounds from the root of Angelica dahurica. AB - The 80% aqueous MeOH extracts from the root of Angelica dahurica, found to inhibit the activities of GABA degradative enzymes GABA transaminase (GABA-T) and succinic semialdehyde dehydrogenase (SSADH), were fractionated using EtOAc, n BuOH and H2O. Repeated column chromatography for the EtOAc and n-BuOH fractions led to the isolation of two new coumarins, oxypeucedanin hydrate-3''-butyl ether and isopraeroside IV along with six known coumarins, isoimperatorin, imperatorin, phellopterin, oxypeucedanin hydrate, nodakenin and 3'-hydroxymarmesinin, and two polyacetylenes, falcarindiol and octadeca-1,9-dien-4,6-diyn-3,8,18-triol. Of the isolated pure compounds, imperatorin and falcarindiol inactivated the GABA-T activities in both time- and concentration-dependent manners. The kinetic studies showed that imperatorin and falcarindiol reacted with the GABA-T with a second order rate constant of 2.3 +/- 0.2 mm(-1) min(-1) and 1.5 +/- 0.1 mm(-1) min(-1), respectively. It is postulated that imperatorin and falcarindiol are able to elevate the neurotransmitter GABA levels in the central nervous system by an inhibitory action on the GABA degradative enzyme GABA-T. PMID- 16261513 TI - Effects of traditional herbal medicine, Hwaotang, on atherosclerosis using the spontaneous familial hypercholesterolemia model, Kurosawa and Kusanagi hypercholesterolemic rabbits and the venous thrombosis rats. AB - Hwaotang (HOT), a traditional Korean medicinal formulation, is a dried decoctum of a mixture of seven herbal medicines, consisting of Angelica gigantis Radix, Rehmanniae Radix, Paeoniae Radix, Ciniamomi Cortex, Cnidii Rhizoma, Persicae Semen and Carthami Flos. In the present study, the inhibitory effects and anti thrombic properties of HOT on the progression of atherosclerotic lesions were studied using the spontaneous familial hypercholesterolemia (FH) model, Kurosawa and Kusanagi-hypercholesterolemic (KHC) rabbits and rats. Changes in blood chemistry, pathology and low-density lipoprotein (LDL) oxidation were measured in a control and HOT group. In the control group, the area of atheromatous plaques of the aorta progressed between week 12 (36.65%) and week 14 (46.22%). This progression of atherosclerotic lesions did not occur in the HOT-treated group after 12 (24.24%) and 14 (23.34%) weeks. Antioxidative effects on LDL were seen in the HOT in weeks 12 and 14. HOT improved the hypercholesterolemia in the KHC rabbits. On the other hand, HOT and five of the seven herbs, except Cnidii Rhizoma and Carthami Flos, inhibited the endotoxin-induced hepatic venous thrombosis in high cholesterol diet-treated rats. However, Ciniamomi Cortex showed a very weak inhibitory effect on the endotoxin-induced hepatic venous thrombosis. The extract also inhibited the endotoxin-induced decrease in blood platelets and fibrinogen, and endotoxin-induced increase in fibrin degradation products (FDP) on disseminated intravascular coagulation in normal rats. In conclusion, these results suggest that HOT has inhibitory effects on the development of atheromatous plaque formation in spontaneous FH rabbits. It is also suggested that the antioxidative effects of HOT on LDL led to the beneficial effects observed in this study. The protection by HOT and its herbs on the artificially induced ischemic infarction might be related to their inhibitory effects on disseminated intravascular coagulation, platelet coagulation and thrombotic action. PMID- 16261514 TI - Hydroalcohol extract and fractions of Stachys lavandulifolia Vahl: effects on spontaneous motor activity and elevated plus-maze behaviour. AB - The present study aimed to investigate the anxiolytic effects of four fractions of Stachys lavandulifolia Vahl. The aerial parts of the plant were extracted with petroleum ether (PF), ethyl acetate (EF), butanol (BF) and water (AF) and tested for spontaneous motor activity and elevated plus-maze (EPM) behaviour in mice. The hydroalcohol extract (HE) and different fractions of S. lavandulifolia were administered intraperitoneally to male Syrian mice, at various doses, 30 min before the behavioural evaluation. The HE of S. lavandulifolia (at 50 mg/kg) increased the percentage of time spent (39%) and the percentage of arm entries in the open arms (53%). The HE (50 mg/kg), PF (25 and 50 mg/kg), EF (25 and 50 mg/kg) and AF (50 mg/kg) of S. lavandulifolia significantly increased the percentage of time spent and the percentage of arm entries in the open arms. The BF up to a dose of 50 mg/kg had no significant effects on any of the measured parameters in the EPM. The spontaneous locomotor activity was significantly decreased in animals injected with each plant fractions, compared with that of saline. The EF and AF showed the least and the most reduction in the activity, respectively. The anxiolytic effects of EF, PF and AF could be related to their content of flavonoids, phenylpropanoids or terpenoids. PMID- 16261515 TI - Evaluation of flavonoids from Bauhinia megalandra leaves as inhibitors of glucose 6-phosphatase system. AB - From the methanol extract of Bauhinia megalandra fresh leaves, eight flavonoids were isolated and evaluated by rat liver microsomal glucose-6-phosphatase (G-6 Pase) bioassay, which might be a useful methodology for screening antihyperglycaemic substances. All the flavonoids assayed showed an inhibitory effect on the intact microsomal G-6-Pase: quercetin and kaempferol exhibited the lowest effect; astilbin, quercetin 3-O-alpha-rhamnoside, kaempferol 3-O-alpha rhamnoside and quercetin 3-O-alpha-arabinoside an intermediate effect. The highest inhibitory activity was shown by quercetin 3-O-alpha-(2'' galloyl)rhamnoside and kaempferol 3-O-alpha-(2''galloyl)rhamnoside. None of the flavonoids mentioned above showed an inhibitory effect on the disrupted microsomal G-6-Pase. Quercetin 3-O-alpha-(2''-galloyl)rhamnoside and kaempferol 3 O-alpha-(2''-galloyl)rhamnoside exhibited the lowest IC50 of all the flavonoids assayed. Also, the phlorizin IC50 is reported. PMID- 16261516 TI - The toxicity of extracts of plant parts of Moringa stenopetala in HEPG2 cells in vitro. AB - The cytotoxicity of extracts from a widely used species of plant, Moringa stenopetala, was assessed in HEPG2 cells, by measuring the leakage of lactate dehydrogenase (LDH) and cell viability. The functional integrity of extract exposed cells was determined by measuring intracellular levels of ATP and glutathione (GSH). The ethanol extracts of leaves and seeds increased significantly (p < 0.01) LDH leakage in a dose- and time-dependent manner. The water extract of leaves and the ethanol extract of the root did not increase LDH leakage. A highly significant (p < 0.001) decrease in HEPG2 viability was found after incubating the cells with the highest concentration (500 microg/mL) of the ethanol leaf and seed extracts. At a concentration of 500 microg/mL, the water extract of leaves increased (p < 0.01), while the ethanol extract of the same plant part decreased (p < 0.01), ATP levels. The root and seed extracts had no significant effect on ATP levels. The ethanol leaf extract decreased GSH levels at a concentration of 500 microg/mL (p < 0.01), as did the ethanol extract of the seeds at 250 microg/mL and 500 microg/mL (p < 0.05). The water extract of the leaves did not alter GSH or LDH levels or affect cell viability, suggesting that it may be non-toxic, and is consistent with its use as a vegetable. The data obtained from the studies with the ethanol extract of the leaves and seeds from Moringa stenopetala show that they contain toxic substances that are extractable with organic solvents or are formed during the process of extraction with these solvents. The significant depletion of ATP and GSH only occurred at concentrations of extract that caused leakage of LDH. Further investigation with this plant in order to identify the constituents extracted and their individual toxic effects both in vivo and in vitro is warranted. This study also illustrates the utility of cell culture for screening plant extracts for potential toxicity. PMID- 16261517 TI - A pilot trial evaluating Meta050, a proprietary combination of reduced iso-alpha acids, rosemary extract and oleanolic acid in patients with arthritis and fibromyalgia. AB - The aim of this open-label, 8-week observational trial was to investigate the efficacy of Meta050 (a proprietary, standardized combination of reduced iso-alpha acids from hops, rosemary extract and oleanolic acid) on pain in patients with rheumatic disease. Osteoarthritis, rheumatoid arthritis and fibromyalgia patients were given 440 mg Meta050 three times a day for 4 weeks, which was changed to 880 mg twice a day for the subsequent 4 weeks in the majority of patients. Pain and condition-specific symptoms were assessed using a standard visual analog scale (VAS), an abridged arthritis impact measurement scale (AIMS2) and the fibromyalgia impact questionnaire. Fifty-four subjects with rheumatic disease completed the trial. Following treatment, a statistically significant decrease in pain of 50% and 40% was observed in arthritis subjects using the VAS (p < 0.0001; Wilcoxon-ranked sums) and AIMS2 (p < 0.0001), respectively. Fibromyalgia subject scores did not significantly improve. A decreasing trend of C-reactive protein, a marker for inflammation, was also observed in those subjects who presented with elevated C-reactive protein. No serious side effects were observed. These observations suggest that Meta050 at a dosage of 440 mg three times a day has a beneficial effect on pain in arthritis subjects. PMID- 16261518 TI - Antitubercular sterols from Thalia multiflora Horkel ex Koernicke. AB - Bioassay guided isolation of an antitubercular extract of the aerial parts of Thalia multiflora led to the isolation of nine stigmast-5-ene and stigmasta-5,22 dien steroids, four isorhamnetin and quercetin flavonoid glycosides, two ceramides, an indole alkaloid and two simple phenolic compounds. Stigmast-5-en 3beta-ol-7-one (2), stigmast-4-ene-6beta-ol-3-one (3), stigmast-5,22-dien-3beta ol-7-one (7) and stigmast-4,22-dien-6beta-ol-3-one (8) were found to be the most active compounds with MIC values of 1.98 +/- 0.02, 4.2 +/- 0.17, 1.0 +/- 0.06 and 2.2 +/- 0.3 microg/mL, respectively. Compounds 2, 3, 7 and 8 were not cytotoxic to Vero cells at 102 microg/mL. This investigation constitutes the first report of a chemical study of a species of the genus Thalia. PMID- 16261519 TI - Hypotensive activity and toxicology of constituents from root bark of Polyalthia longifolia var. pendula. AB - A defatted extract of Polyalthia longifolia var. pendula root bark (PRB) in 50% methanol showed a significant ability to reduce blood pressure. It caused a 22% and 47% fall in mean arterial blood pressure (MABP) in rats at doses of 3 mg/kg and 30 mg/kg, respectively. Compounds purified from this extract include kolavenic acid (3), clerodane (1) and its isomer (2), liriodenine (4), lysicamine (5) and bisclerodane imide (6) and its isomer (7). Of these, only kolavenic acid produced a 22% fall in MABP, at a dose of 30 mg/kg. PRB showed a decrease in blood pressure of normotensive and egg yolk induced hypertensive rats. The LD50 of PRB was determined as 100 mg/kg in mice. PMID- 16261520 TI - Development of pregnancy in rats treated with Hypericum perforatum. AB - St John's wort (Hypericum perforatum) is a medicinal plant used in the treatment of depression and other psychiatric disorders. In the present paper, the toxicity of H. perforatum administered to female rats during the period of organogenesis (day 9-15 of pregnancy) was evaluated. Thirty inseminated Wistar rats were randomly distributed into control and treated groups, which received, by gavage, 0.5 mL of saline and 36 mg/kg body weight of Jarsin dried extract diluted into 0.5 mL of saline, respectively. Maternal toxicity was evaluated through: water and food intake, body weight gain, piloerection, locomotor activity, diarrhea and death occurrence. Animals were killed on day 21 of pregnancy, when fetuses and placentas were removed and weighed. The indices of implantation and resorption were calculated. Clinical signs of maternal toxicity were not observed and none of the variables analysed showed statistically significant differences. In the dose administered in the experimental model used, H. perforatum does not seem to be toxic to the mother nor to interfere with the progress of gestation during organogenesis. PMID- 16261521 TI - Evaluation of the antibacterial activity of Ventilago madraspatana Gaertn., Rubia cordifolia Linn. and Lantana camara Linn.: isolation of emodin and physcion as active antibacterial agents. AB - The antibacterial activity of the extracts of Ventilago madraspatana stem-bark, Rubia cordifolia root and Lantana camara root-bark, prepared with solvents of different polarity, was evaluated by the agar-well diffusion method. Twelve bacteria, six each of gram-positive and gram-negative strains, were used in this study. Chloroform and ethanol extracts of V. madraspatana showed broad-spectrum activity against most of the bacteria except S. aureus, E. coli and V. cholerae. On the other hand, the activity of the chloroform and methanol extracts of R. cordifolia and L. camara was found to be more specific towards the gram-positive strains, although gram-negative P. aeruginosa was also inhibited by the methanol extracts of both these plants in a dose dependent manner. The water extracts of V. madraspatana and L. camara were found to be inactive, while that of R. cordifolia was significantly active against B. subtilis and S. aureus compared with streptomycin and penicillin G used as standards. In the course of bio-assay guided fractionation, emodin and physcion were isolated for the first time from the stem-bark of V. madraspatana. It was noteworthy to find the MICs of emodin in the range 0.5-2.0 microg/mL against three Bacillus sp. Both the anthraquinonoid compounds inhibited P. aeruginosa, emodin being more effective, showing an MIC of 70 microg/mL. PMID- 16261522 TI - Haemolytic activities and adjuvant effect of Gynostemma pentaphyllum saponins on the immune responses to ovalbumin in mice. AB - In this study, the haemolytic activities of Gynostemma pentaphyllum saponins (GPS) and its adjuvant potential on the immune responses of ICR mice against ovalbumin (OVA) were evaluated. GPS showed a slight haemolytic effect, with its haemolytic activity being 10.20% and 4.90% at concentrations of 500 and 250 microg/mL, respectively. ICR mice were immunized subcutaneously with OVA 100 microg alone or with OVA 100 microg dissolved in saline containing aluminium hydroxide gel (Alum, 200 microg), QuilA (10 and 20 microg) or GPS (50, 100 or 200 microg) on days 1 and 15. Two weeks later (day 28), concanavalin A (Con A)-, lipopolysaccharide (LPS)- and OVA-stimulated splenocyte proliferation and OVA specific antibodies in serum were measured. GPS significantly enhanced the Con A , LPS- and OVA-induced splenocyte proliferation in the OVA-immunized mice, especially at a dose of 100 microg (p < 0.05 or p < 0.001). OVA-specific IgG, IgG1 and IgG2b antibody levels in serum were also significantly enhanced by GPS compared with the OVA control group (p < 0.05, p < 0.01 or p < 0.001). In conclusion, the results suggest that GPS could be used safely as an adjuvant with low or no haemolytic effect. PMID- 16261523 TI - Effects of St John's wort (Hypericum perforatum L.) extract on plasma androgen concentrations in healthy men and women: a pilot study. AB - St John's wort extract (SJW; Hypericum perforatum L.) is taken extensively as a putative herbal antidepressant. It has been shown to induce the activity of cytochrome P-450 3A4 (CYP3A4) and to increase the clearance of numerous drugs and steroids such as cortisol and ethinyl estradiol. This study was conducted to determine if SJW exposure also alters the concentrations of circulating androgenic steroid hormones. The study was conducted using healthy volunteers (6M, 6F) studied before and after a 14-day treatment period with a SJW preparation previously demonstrated to induce the activity of CYP3A4. Plasma concentrations of testosterone, dihydrotestosterone (DHT), dehydroepiandrosterone sulfate (DHEAS), sex hormone-binding globulin (SHBG) and the combined concentrations of androsterone sulfate (AoS) and epiandrosterone sulfate (epiAoS) were measured by immunoassay methods. The results of analysis demonstrated that SJW did not significantly alter the majority of the androgens studied (p > 0.05) although the combined concentrations of the 5alpha-reduced steroids, AoS and epiAoS, significantly declined following treatment in all subjects (p = 0.02), and in males (p = 0.04). Furthermore, the testosterone to DHT ratio was increased in both men and women. Although the latter increase did not reach statistical significance, it is also consistent with the possible inhibition of 5alpha reductase by SJW. It is concluded that despite significant induction of CYP3A4, short term administration of SJW does not significantly alter the concentrations of most circulating androgens in men and women but may produce a dimunition in some of the circulating 5alpha-reduced androgens. PMID- 16261524 TI - Antioxidant activity of five vegetables traditionally consumed by South-Asian migrants in Bradford, Yorkshire, UK. AB - Five vegetables traditionally consumed among South-Asian migrants in Bradford (Yorkshire, UK) were tested for their free radical scavenging activity (FRSA) in the DPPH (1,1-diphenyl-2-picrylhydrazil radical) screening assay (using extracts prepared both by cold maceration and also by boiling the plant in the solvent under reflux) and for their in vitro non-enzymatic inhibition of bovine brain lipid peroxidation. In both antioxidant assays a strong activity was shown by extracts derived from okra (Abelmoschus esculentus, Malvaceae) fruits and charungli (Caralluma edulis, Asclepiadaceae) aerial parts. Extracts from bitter melon (Momordica charantia, Cucurbitaceae) and angular loofah (Luffa acutangula) showed a significant difference in the FRSA between the extract obtained by using cold maceration and that prepared by boiling the plant in the solvent under reflux, suggesting the chemical composition of the plant changed during the heating process, leading to an increase in the amount of antioxidant components. These findings confirm the great interest of the nutraceutical sciences in extracts of Caralluma edulis, whose phytochemistry and phytopharmacology should be investigated further in order to detect possible phytotherapeutic uses in the prevention of ageing related diseases (ARDs) and Alzheimer disease (AD). PMID- 16261525 TI - Protective effects of lycopene and tomato extract against doxorubicin-induced cardiotoxicity. AB - The protective effect of tomato extract and lycopene on acute doxorubicin (DOX) myocardial toxicity was evaluated in mice. DOX toxicity, induced by a single intraperitoneal injection (15 mg/kg), was revealed by an elevated serum CPK(MB) and histopathological observations. Tomato extract (1.2 and 2.4 g/kg, i.p.) and lycopene (1.7 and 3.5 mg/kg, i.p.) prevented the rise in serum CPK(MB) and ameliorated cardiac cell injury. These results suggest that tomato extract and lycopene inhibit DOX cardiotoxicity and might serve as a novel combination chemotherapeutic agent with DOX to limit free radical-mediated organ injury. PMID- 16261526 TI - A Kampo formula Juzen-taiho-to induces expression of metallothioneins in mice. AB - Juzen-taiho-to (JTX), one of the commonly prescribed traditional Japanese herbal medicines (Kampo), is indicated for adjunctive treatment of cancers and autoimmune diseases. To understand the mechanisms underlying the clinical effects of JTX, the effects of orally administered JTX on the expression of metallothioneins (MTs) were examined in the liver, spleen, small and large intestines of mice. In addition, the expression of MTs in specific pathogen free (SPF) mice was examined to understand the participation of intestinal bacteria in the expression of MTs. JTX enhanced expression of MT-I and -II significantly in the liver of SPF mice. Induction of MT-II expression was observed also in the small intestine. Intestinal bacteria appeared to have no effect on MTs expression. Neither expression of MT-III nor its induction was observed in any tissue. These findings strongly suggest that MTs should mediate at least some effects of JTX in mice. PMID- 16261527 TI - Amelioration of non-alcoholic steatohepatitis and glucose intolerance in ob/ob mice by oral immune regulation towards liver-extracted proteins is associated with elevated intrahepatic NKT lymphocytes and serum IL-10 levels. AB - Non-alcoholic steatohepatitis (NASH) is a common cause of cryptogenic cirrhosis in the Western world. In an animal model of NASH, leptin-deficient ob/ob mice present with alterations in number and function of hepatic NKT and peripheral CD4 lymphocytes. Oral immune regulation is a method to alter the immune response towards orally administered antigens. To determine the effect of oral immune regulation towards liver-extracted proteins on the metabolic disorders in ob/ob mice, ob/ob mice and their lean littermates were orally administered liver extracts from wild-type or ob/ob mice or bovine serum albumin for 1 month. The effect of treatment on hepatic fat content was measured by magnetic resonance imaging (MRI) and using a histological steatohepatitis grading scale. Glucose tolerance was measured by an oral glucose tolerance test (GTT). T lymphocyte subpopulations were assessed by flow cytometry analysis. Induction of immune regulation by oral presentation of liver-extracted proteins resulted in a significant 18% reduction of the hepatic fat content in ob/ob mice fed with either wild-type or ob/ob liver extracts for 1 month. The MRI signal intensity index in treated mice decreased to 0.48 and 0.51, respectively, compared with 0.62 in BSA-fed controls (p = 0.037 and p = 0.019, respectively), while the histological steatohepatitis score decreased in both treated groups to 2.0, compared with 2.4 in BSA-fed controls (p = 0.05). A significant improvement in GTT was noted in treated ob/ob mice. These changes were accompanied by a marked increase in the intrahepatic NKT lymphocyte population in mice fed with proteins extracted from both wild-type and ob/ob mice (46.96% and 56.72%, respectively, compared with 26.21% in BSA-fed controls; p < 0.05) and a significant elevation in serum IL-10 levels. Oral immune regulation towards liver extracted proteins in leptin-deficient mice resulted in a marked reduction in hepatic fat content and improved glucose tolerance. This effect was associated with a significant increase in the intrahepatic NKT lymphocyte population and serum IL-10 levels, suggesting a Th1 to Th2 immune shift. Immune regulation towards disease associated antigens holds promise as a new mode of therapy for NASH. PMID- 16261528 TI - Central projections of auditory receptor neurons of crickets. AB - We describe the central projections of physiologically characterized auditory receptor neurons of crickets as revealed by confocal microscopy. Receptors tuned to ultrasonic frequencies (similar to those produced by echolocating, insectivorous bats), to a mid-range of frequencies, and a subset of those tuned to low, cricket-like frequencies have similar projections, terminating medially within the auditory neuropile. Quantitative analysis shows that despite the general similarity of these projections they are tonotopic, with receptors tuned to lower frequencies terminating more medially. Another subset of cricket-song tuned receptors projects more laterally and posteriorly than the other types. Double-fills of receptors and identified interneurons show that the three medially projecting receptor types are anatomically well positioned to provide monosynaptic input to interneurons that relay auditory information to the brain and to interneurons that modify this ascending information. The more laterally and posteriorly branching receptor type may not interact directly with this ascending pathway, but is well positioned to provide direct input to an interneuron that carries auditory information to more posterior ganglia. These results suggest that information about cricket song is segregated into functionally different pathways as early as the level of receptor neurons. Ultrasound-tuned and mid-frequency tuned receptors have approximately twice as many varicosities, which are sites of transmitter release, per receptor as either anatomical type of cricket-song-tuned receptor. This may compensate in part for the numerical under-representation of these receptor types. PMID- 16261529 TI - Gene expression differences over a critical period of afferent-dependent neuron survival in the mouse auditory brainstem. AB - Deprivation of auditory nerve input in young mice results in dramatic neuron death in the anteroventral cochlear nucleus (CN), while the same manipulation performed in older mice does not result in significant neuronal loss. The molecular basis underlying this critical period of susceptibility to loss of afferent input remains largely unknown. One possibility is that developmental differences in baseline mRNA expression of specific genes could predispose CN neurons to either death or survival after deafferentation. We used two microarray platforms to identify differentially expressed genes in the CN during and after this critical period. Results across platforms were also compared to each other. Three ages were examined; during the critical period (postnatal day (P)7), at the closing of the critical period (P14), and 1 week after the critical period (P21). Of all the genes surveyed (22,690 or 15,247), 1,082 were identified as significantly changed in expression during the critical period relative to after. Real-time reverse transcription polymerase chain reaction and immunohistochemistry confirmed and extended the microarray results for a subset of these genes. Further analysis of genes related to apoptotic pathways showed 6 out of 7 differentially expressed known pro-apoptotic genes had higher expression during the critical period. In contrast, 9 out of 11 differentially expressed known pro-survival genes increased after the critical period when CN neurons survive deprivation. This finding supports the concept that multiple neuroprotective mechanisms increase and pro-apoptotic factors decrease over development to protect mature neurons from stressful insults, making them less dependent on afferent input for survival. PMID- 16261531 TI - DNA topoisomerase I in the mouse central nervous system: Age and sex dependence. AB - Topoisomerase I (topo I) is a nuclear enzyme responsible for the topological state of DNA and therefore participates in most DNA transactions, particularly in transcription. Topo I, a ubiquitous enzyme, was identified and characterized in various cell types and tissues; however, the characterization of topo I in the intact central nervous system was not performed. Here we investigated, for the first time, the activity, level, and distribution pattern of topo I in the various selected brain regions in the mouse. In the visual cortex, cerebellum, and striatum the activity of topo I was 3-4-fold higher compared to that found in the hippocampus and hypothalamus. Immunohistochemical and immunofluorescence analyses revealed specific distribution patterns of topo I protein in neurons of each of the areas examined. The highest topo I levels were observed in inhibitory neurons. In addition to the expected nuclear localization of this protein, some neurons exhibited significant cytoplasmic content as well. The activity and level of topo I is age- and gender-dependent. It increases from birth to maturity and decreases, more significantly in males, with senescence. These results point to a possible importance and involvement of topo I activity and regulation in various brain functions. PMID- 16261530 TI - Neuron-to-astrocyte transition: phenotypic fluidity and the formation of hybrid asterons in differentiating neurospheres. AB - To the extent that their fate choice and differentiation processes can be understood and manipulated, neural stem cells represent a promising therapeutic tool for a variety of neuropathologies. We have previously shown that mature astrocytes possess neural stem cell attributes, and can give rise to neurons through the formation of multipotent neurosphere clones. Here we show that relatively mature neurons generated from neurospheres derived from postnatal subependymal zone or cerebellar cortex undergo a phenotypic transformation into astrocytes that coincides with the appearance of a nonfused, hybrid cell type that shares the morphology, antigenicity, and physiology of both neurons and astrocytes. We refer to this astrocyte/neuron hybrid as an "asteron," and hypothesize that it represents an intermediate step in the trans- or dedifferentiation of neurons into astrocytes. The present finding suggests that seemingly terminally differentiated neural cells may in fact represent points along a bidirectionally fluid continuum of differentiation, with intermediate points represented by "hybrid" cells coexpressing phenotypic markers of more than one lineage. PMID- 16261532 TI - Modulation of intercellular calcium signaling by melatonin in avian and mammalian astrocytes is brain region-specific. AB - Calcium waves among glial cells impact many central nervous system functions, including neural integration and brain metabolism. Here, we characterized the modulatory effects of melatonin, a pineal neurohormone that mediates circadian and seasonal processes, on glial calcium waves derived from different brain regions and species. Diencephalic and telencephalic astrocytes, from both chick and mouse brains, expressed melatonin receptor proteins. Further, using the calcium-sensitive dye Fluo-4, we conducted real-time imaging analyses of calcium waves propagated among mammalian and avian astrocytes. Mouse diencephalic astrocytic calcium waves spread to an area 2-5-fold larger than waves among avian astrocytes and application of 10 nM melatonin caused a 32% increase in the spread of these mammalian calcium waves, similar to the 23% increase observed in chick diencephalic astrocytes. In contrast, melatonin had no effect on calcium waves in either avian or mammalian telencephalic astrocytes. Mouse telencephalic calcium waves radially spread from their initiation site among untreated astrocytes. However, waves meandered among mouse diencephalic astrocytes, taking heterogeneous paths at variable rates of propagation. Brain regional differences in wave propagation were abolished by melatonin, as diencephalic astrocytes acquired more telencephalon-like wave characteristics. Astrocytes cultured from different brain regions, therefore, possess fundamentally disparate mechanisms of calcium wave propagation and responses to melatonin. These results suggest multiple roles for melatonin receptors in the regulation of astroglial function, impacting specific brain regions differentially. PMID- 16261533 TI - Organization of optic lobes that support motion detection in a semiterrestrial crab. AB - There is a mismatch between the documentation of the visually guided behaviors and visual physiology of decapods (Malacostraca, Crustacea) and knowledge about the neural architecture of their visual systems. The present study provides a description of the neuroanatomical features of the four visual neuropils of the grapsid crab Chasmagnathus granulatus, which is currently used as a model for investigating the neurobiology of learning and memory. Visual memory in Chasmagnathus is thought to be driven from within deep retinotopic neuropil by large-field motion-sensitive neurons. Here we describe the neural architecture characterizing the Chasmagnathus lobula, in which such neurons are found. It is shown that, unlike the equivalent region of insects, the malacostracan lobula is densely packed with columns, the spacing of which is the same as that of retinotopic units of the lamina. The lobula comprises many levels of strata and columnar afferents that supply systems of tangential neurons. Two of these, which are known to respond to movement across the retina, have orthogonally arranged dendritic fields deep in the lobula. They also show evidence of dye coupling. We discuss the significance of commonalties across taxa with respect to the organization of the lamina and medulla and contrasts these with possible taxon specific arrangements of deeper neuropils that support systems of matched filters. PMID- 16261534 TI - Projections from the subfornical region of the lateral hypothalamic area. AB - The L-shaped anterior zone of the lateral hypothalamic area's subfornical region (LHAsfa) is delineated by a pontine nucleus incertus input. Functional evidence suggests that the subfornical region and nucleus incertus modulate foraging and defensive behaviors, although subfornical region connections are poorly understood. A high-resolution Phaseolus vulgaris-leucoagglutinin (PHAL) structural analysis is presented here of the LHAsfa neuron population's overall axonal projection pattern. The strongest LHAsfa targets are in the interbrain and cerebral hemisphere. The former include inputs to anterior hypothalamic nucleus, dorsomedial part of the ventromedial nucleus, and ventral region of the dorsal premammillary nucleus (defensive behavior control system components), and to lateral habenula and dorsal region of the dorsal premammillary nucleus (foraging behavior control system components). The latter include massive inputs to lateral and medial septal nuclei (septo-hippocampal system components), and inputs to bed nuclei of the stria terminalis posterior division related to the defensive behavior system, intercalated amygdalar nucleus (projecting to central amygdalar nucleus), and posterior part of the basomedial amygdalar nucleus. LHAsfa vertical and horizontal limb basic projection patterns are similar, although each preferentially innervates certain terminal fields. Lateral hypothalamic area regions immediately medial, lateral, and caudal to the LHAsfa each generate quite distinct projection patterns. Combined with previous evidence that major sources of LHAsfa neural inputs include the parabrachial nucleus (nociceptive information), defensive and foraging behavior system components, and the septo hippocampal system, the present results suggest that the LHAsfa helps match adaptive behavioral responses (either defensive or foraging) to current internal motivational status and external environmental conditions. PMID- 16261537 TI - Precentral sulcal complex of the human brain: morphology and statistical probability maps. AB - The morphology of the precentral sulcus was examined via 3D visualization in 40 structural magnetic resonance images of the human brain to define its common features and their variability. The precentral sulcus is composed of two distinct sulcal configurations: 1) the inferior precentral sulcus (IP), situated caudal to the inferior frontal sulcus, and 2) the superior precentral sulcus (SP), caudal to the superior frontal sulcus. The SP was usually a single connected structure, and only in 24% of cases studied did it consist of two separate folds. The caudal end of the superior frontal sulcus and the SP merge on the surface of the brain. However, a clear separation can be established in 72% of cases by analyzing the morphology of the depth of this region in serial sections. Two or three small sulci can be identified in the area between the SP and the midline: the medial precentral sulcus, the marginal precentral sulcus, and a paramidline sulcus. The IP is composed of three different parts: a dorsal and a ventral branch, both oriented vertically, and a short horizontal extension. The ventral branch forms the caudal border of the inferior frontal gyrus, the dorsal branch forms the caudal border of the ventral portion of the middle frontal gyrus, and the horizontal extension runs in a rostrodorsal direction into the middle frontal gyrus. These three structures are closely related to each other but can, in most cases, be separated in the depth of the inferior precentral sulcal complex. PMID- 16261538 TI - Localization of 5alpha-reductase in the rat main olfactory bulb. AB - The enzyme steroid 5alpha-reductase catalyzes the production of dihydroprogesterone and dihydrotestosterone, which were recently recognized as neurosteroids in the brain with variably potential neuroactivity. The present study reports for the first time detailed localization of 5alpha-reductase type 1 in the rat main olfactory bulb. The occurrence of 5alpha-reductase in the olfactory bulb was detected by reverse transcription-polymerase chain reaction and Western blotting analyses. In addition, the enzyme activity was also detected by thin layer chromatography. Immunocytochemistry showed that 5alpha-reductase immunoreactive cells of variable intensity were present in all layers of the olfactory bulb. Multiple immunolabeling revealed that 5alpha-reductase was mainly localized in glial cells, namely, in S-100beta- and glial fibrillary acidic protein-immunoreactive astrocytes, 2', 3'-cyclic nucleotide 3'-phosphodiesterase (CNPase)-immunoreactive oligodendrocytes, and in S-100beta- and neuropeptide-Y immunoreactive olfactory ensheathing cells, whereas the bulbar neurons exhibited little immunoreactivity. Quantitative analysis revealed that the number of 5alpha reductase-immunoreactive cells was greatest in the olfactory nerve layer. The most intense 5alpha-reductase-immunoreactivity was found in the olfactory ensheathing cells, and next in the CNPase-immunoreactive cells. The 5alpha reductase in the olfactory bulb was expressed constantly throughout different ages and sexes and in neutered and hypophysectomized rats. Thus, 5alpha-reductase may contribute via 5alpha-reduced metabolites to the formation and maintenance of olfactory inputs and outputs, which were closely associated with the olfactory ensheathing cells and the oligodendrocytes, respectively. PMID- 16261535 TI - Compartmental localization of gamma-aminobutyric acid type B receptors in the cholinergic circuitry of the rabbit retina. AB - Although many effects of gamma-aminobutyric acid (GABA) on retinal function have been attributed to GABA(A) and GABA(C) receptors, specific retinal functions have also been shown to be mediated by GABA(B) receptors, including facilitation of light-evoked acetylcholine release from the rabbit retina (Neal and Cunningham [1995] J. Physiol. 482:363-372). To explain the results of a rich set of experiments, Neal and Cunningham proposed a model for this facilitation. In this model, GABA(B) receptor-mediated inhibition of glycinergic cells would reduce their own inhibition of cholinergic cells. In turn, muscarinic input from the latter to the glycinergic cells would complete a negative-feedback circuitry. In this study, we have used immunohistochemical techniques to test elements of this model. We report that glycinergic amacrine cells are GABA(B) receptor negative. In contrast, our data reveal the localization of GABA(B) receptors on cholinergic/GABAergic starburst amacrine cells. High-resolution localization of GABA(B) receptors on starburst amacrine cells shows that they are discretely localized to a limited population of its varicosities, the majority of likely synaptic-release sites being devoid of detectable levels of GABA(B) receptors. Finally, we identify a glycinergic cell that is a potential muscarinic receptor bearing target of GABA(B)-modulated acetylcholine release. This target is the DAPI-3 cell. We propose, based on these data, a modification of the Neal and Cunningham model in which GABA(B) receptors are on starburst, not glycinergic amacrine cells. PMID- 16261539 TI - Contiguous spread of Mycobacterium ulcerans in Buruli ulcer lesions analysed by histopathology and real-time PCR quantification of mycobacterial DNA. AB - The distribution of M. ulcerans in Buruli ulcer lesions was analysed by IS2404 real-time PCR quantification of M. ulcerans DNA and by semi-quantitative microscopic assessment of the number of acid-fast bacilli (AFB). Mycobacterial burden was compared with histopathological changes. Focal distribution of tissue destruction extending into areas with high and low mycobacterial burden was a feature in all lesions analysed. Even where most of the mycobacteria were washed out of ulcerative lesions, peaks of mycobacterial DNA and AFB in the necrotic base of the ulcers still marked the position of the primary infection focus. Significant amounts of mycobacterial DNA and microcolonies were also present in samples from more peripheral regions and occasionally in excised margins of macroscopically and histologically healthy-appearing excised tissue margins. Additional peaks of mycobacterial DNA clearly marked sites where satellite lesions were developing. Even when granulomas provided evidence for the development of cell-mediated immunity, development of satellite lesions by contiguous spreading was not completely prevented. Areas free of mycobacterial DNA were found between primary and secondary infection foci and around scarring tissue of healing lesions. These results demonstrate that IS2404 real-time PCR analysis is a better tool than the less sensitive and only semi-quantitative microscopic enumeration of AFB for studying the dynamics of M. ulcerans infection in situ. PMID- 16261540 TI - Simulation of mefenacet concentrations in paddy fields by an improved PCPF-1 model. AB - An improved simulation model (PCPF-1) has been evaluated for the prediction of the fate of mefenacet in an experimental paddy field. This model simulates the fate and transport of pesticide in paddy water and the top 1 cm of paddy soil. Observed concentrations of mefenacet in the paddy water and the surface soil exponentially decreased from their maximum concentrations of 0.70 mg litre(-1) and 11.3 mg kg(-1), respectively. Predicted mefenacet concentrations both in the water and surface soil were in excellent agreement with those measured during the first 2 weeks after herbicide application, but concentrations in paddy water were appreciably overestimated thereafter. The model simulated mefenacet losses through runoff, percolation and degradation to be respectively 41.9%, 6.4% and 57.3% of applied, and the mass balance error was about -6%. The model simulation implied that drainage and seepage control, especially shortly after application when herbicide concentrations are high, is essential for preventing pesticide losses from paddy fields. In focusing on pesticide concentrations in this early period the PCPF-1 model can be a beneficial tool for risk assessment of pesticide losses and in the evaluation of agricultural management for reducing pesticide pollution associated with paddy rice production. PMID- 16261541 TI - Acceptability of pesticide impacts on the environment: what do United Kingdom stakeholders and the public value? AB - EU Directive 91/414/EEC requires there to be no unacceptable effects on the environment from the use of pesticides. This paper reports the views of direct stakeholder groups and results from an opinion survey of more than 2000 members of the general public on what in practice should constitute acceptable and unacceptable effects of pesticides. Stakeholders in focus groups were concerned with the potential effects of pesticides on animal and plant population viability and micro-organism function but recognized that a trade-off exists between the potential economic advantages of responsible pesticide use and the potential disadvantages of individual poisoning events. The public opinion survey showed that although pesticides are widely used in homes and gardens, their use on farm crops remains of concern to the public. Concerns are greatest on issues of human health and food quality but potential environmental effects are also an issue for a substantial number of people, particularly if attractive species could be affected. PMID- 16261542 TI - Impact of cypermethrin on feeding behaviour and mortality of the spider Pardosa amentata in arenas with artificial 'vegetation'. AB - Pesticides can modify invertebrate movement and feeding behaviour which could reduce predation in agroecosystems. Previous assays have exposed the spider Pardosa amentata (Clerck) to the synthetic pyrethroid cypermethrin and monitored prey items consumed in small containers (requiring very little movement to capture prey). The current study used larger arenas containing artificial 'vegetation' (a plastic analogue) to encourage spiders to hunt and capture prey. The period 24 h after exposure produced greatest variability in prey item consumption between treatments and was used to examine treatment effects. At this time, cypermethrin reduced prey consumption rates but these effects did not persist. Findings did not suggest that the presence of artificial vegetation in arenas modified prey consumption rates, which was consistent for individuals treated with cypermethrin and a control group. This is despite the majority of pesticide-treated individuals exhibiting both ataxia and paralysis of the hind legs (these effects persisting for a maximum of 3 and 6 days respectively). These findings were consistent for both sexes. Spider longevity under starvation conditions was not significantly reduced by cypermethrin exposure but overall females survived longer than males. The findings are discussed in the context of the arenas used and the ecology of this common predator. PMID- 16261544 TI - Rapid determination of haloperidol and its metabolites in human plasma by HPLC using monolithic silica column and solid-phase extraction. AB - A rapid, sensitive and reproducible HPLC method was developed and validated for the analysis of haloperidol and its three main metabolites in human plasma. The analysis was carried out on a monolithic silica column (Chromolith Performance RP 18e, 100 x 4.6 mm). The mobile phase consisted of sodium phosphate (0.1 m, pH 3.5)-acetonitrile (80:20, v/v) at a flow rate of 2.0 mL/min. UV detection at 230 nm was used, with the detection limits of these compounds ranging from 2 to 5 ng. The separation factors of all studied compounds were in the range 2.30-16.32, while the resolution factors were from 1.00 to 5.37. PMID- 16261545 TI - Simple and rapid high-performance liquid chromatographic method for endogenous alpha-tocopherol determination in human plasma. AB - A simple and rapid reversed-phase high-performance liquid chromatographic method was developed and validated for the determination of endogenous alpha-tocopherol in human plasma. Following addition of alpha-tocopheryl acetate as the internal standard, the plasma was deproteinized using acetonitrile and isopropanol mixture prior to HPLC analysis. Methanol was used as the mobile phase and the effluent was quantitated at 292 nm. By this developed method, the concentrations of alpha tocopherol were linearly related to their responses in the range of 0.8-30 microg/mL. The relative standard deviations intra-day and inter-day for alpha tocopherol in plasma were less than 10%. The percentage of bias was within +/-4%, which confirmed the accuracy of the method. The method has been successfully applied for determining endogenous alpha-tocopherol in healthy Thai male volunteers. PMID- 16261546 TI - Implications of the "watermelon seeding" phenomenon during coronary interventions for in-stent restenosis. AB - The occurrence of balloon slippage ("watermelon seeding"; WMS) during treatment of patients with in-stent restenosis (ISR) has been described, but predisposing factors and the potential implications of this phenomenon remain unknown. In the Restenosis Intrastent: Balloon Angioplasty vs. Elective Stenting (RIBS) randomized study, 450 patients with ISR were included. Of these, 42 patients (9%) presented WMS during the procedure. WMS was detected in 26 patients (12%) in the balloon arm and 16 (7%) in the stent arm (P=0.11). In the stent arm, WMS was only noticed during balloon predilation, never during stent implantation. As compared with 408 patients without WMS, patients with WMS had more severe (TIMI flow 1; 21% vs. 8%; P=0.01) and diffuse (length>15 mm: 45% vs. 28%; P=0.02) ISR lesions. Patients with WMS required more balloon inflations, longer total inflation time, had more frequent crossover to stenting or ended the procedure with residual dissections, and eventually obtained poorer acute results (minimal lumen diameter, 2.35+/-0.5 vs. 2.53+/-0.5 mm; P=0.03). In addition, at 6-month follow up, patients with WMS had a smaller minimal lumen diameter (1.26+/-0.7 vs. 1.61+/ 0.7 mm; P=0.007) and a higher restenosis rate (56% vs. 37%; P=0.017). On logistic regression analysis, the WMS phenomenon emerged as an independent predictor of recurrent restenosis (adjusted RR=2.1; 95% CI=1.1-4.1; P=0.04). The WMS phenomenon may complicate treatment of patients with ISR. Long and severe lesions appear to predispose to this technical problem that never occurs during stent deployment. In patients with ISR, WMS is associated with cumbersome procedures and poorer acute and long-term angiographic results. PMID- 16261547 TI - Population structure inferred by local spatial autocorrelation: an example from an Amerindian tribal population. AB - Spatial autocorrelation (SA) methods were recently extended to detect local spatial autocorrelation (LSA) at individual localities. LSA statistics serve as useful indicators of local genetic population structure. We applied this method to 15 allele frequencies from 43 villages of a South American tribe, the Yanomama. Based on a network of links T mutation in exon 12 of COCH that co-segregates with auditory dysfunction in the pedigree. The mutation results in a predicted p.C542F substitution at an evolutionarily conserved cysteine residue in the C-terminus of cochlin. The c.1625G > T transversion in COCH exon 12 represents the first reported mutation outside of the LCCL domain which is encoded by exons 4 and 5. The 542F mutant cochlin is translated and secreted by transfected mammalian cells. Western blot analysis under non-reducing and reducing conditions suggests that the 542F mutation alters intramolecular cochlin disulfide bond formation. In the vestibular system, a progressive horizontal canal hypofunction and a probable saccular otolith challenge were detected in family members with the c.1625G > T COCH alteration. Abnormal central oculomotor test results in family members with the c.1625G > T COCH alteration imply a possible central nervous system change not previously noted in DFNA9 pedigrees harboring mutations within the LCCL domain. PMID- 16261628 TI - Second hit deletion size in von Hippel-Lindau disease. AB - von Hippel-Lindau (VHL) disease is caused by germline mutations of the VHL tumor suppressor gene. Affected individuals are predisposed to development of multiple neoplastic and preneoplastic lesions of different organs. A "second hit," which is usually represented by deletion in the wild-type VHL allele at 3p25, is necessary for initiation of tumor formation. The impact of the size of this deletion is thought to be critical, because other tumor suppressor genes are located nearby, but this has not yet been studied in detail. We mapped the deletion size of the "second hit" in microdissected tissue from 16 different VHL associated lesions from the same patient using 10 polymorphic chromosome 3 markers. We found that the deletion size is highly variable, ranging from short deletions around the VHL gene to complete loss of the chromosome 3. The deletion sizes are not correlated with the site of the germline mutation, the affected organ, or the type or biological behavior of the tumor. Even preneoplastic cystic structures may harbor entire loss of the chromosome 3, suggesting that loss of VHL gene function alone is not immediately causative for neoplastic growth, but further events, either mutations and deletions in other chromosomes or epigenetic or other than genetic phenomena, are required for tumor formation. PMID- 16261629 TI - A new interpretation of ASM2d for modeling of SBR performance for enhanced biological phosphorus removal under different P/HAc ratios. AB - This study evaluated the prediction capability of Activated Sludge Model No. 2d (ASM2d), for the enhanced biological phosphorus removal (EBPR) performance of a sequencing batch reactor (SBR) receiving variable influent phosphate load. For this purpose, a laboratory-scale SBR was operated with a synthetic feed containing acetate as the sole carbon source. The experiments were conducted in four different Runs to ensure a range of different phosphate/acetate ratios in the influent. Model evaluations were carried out using concentration profiles measured throughout a representative cycle at steady state. An iterative calibration methodology was developed based on sensitivity analysis and applied to four different sets of experimental data on relevant model parameters reflecting SBR performance. ASM2d was able to predict the steady state behavior of the SBR system receiving variable influent phosphate loads only with the recalibrated parameter set. The regular changing pattern of the coefficients could be interpreted with the ability of the SBR system to sustain glycogen accumulating microorganisms, GAOs, which can store substrate under anaerobic conditions without polyphosphate energy, but deriving energy from the degradation of glycogen. Thus they are capable of prevailing at lower P/Ac ratios. The results indicate the need to include glycogen and GAOs as model components for processes involving both phosphate accumulating organisms, (PAOs) and GAOs, in order to obtain a better prediction of X(PHA) and oxygen uptake rate (OUR) profiles in the system. PMID- 16261630 TI - On-column refolding of recombinant human interferon-gamma inclusion bodies by expanded bed adsorption chromatography. AB - A refolding strategy was described for on-column refolding of recombinant human interferon-gamma (rhIFN-gamma) inclusion bodies by expanded bed adsorption (EBA) chromatography. After the denatured rhIFN-gamma protein bound onto the cation exchanger of STREAMLINE SP, the refolding process was performed in expanded bed by gradually decreasing the concentration of urea in the buffer and the refolded rhIFN-gamma protein was recovered by the elution in packed bed mode. It was demonstrated that the denatured rhIFN-gamma protein could be efficiently refolded by this method with high yield. Under appropriate experimental conditions, the protein yield and specific activity of rhIFN-gamma was up to 52.7% and 8.18 x 10(6) IU/mg, respectively. PMID- 16261631 TI - A novel superporous agarose medium for high-speed protein chromatography. AB - A novel superporous agarose (SA) bead characterized by the presence of wide pores has been fabricated by water-in-oil emulsification using solid granules of calcium carbonate as porogenic agent. After cross-linking, the solid granules were removed by dissolving them in hydrochloric acid. Then, the gel was modified with diethylaminoethyl groups to create an anion exchanger, SA-DEAE, for protein adsorption. A homogeneous agarose (HA) bead was also produced and modified with DEAE for comparison. It was found that the porosity of SA-DEAE was about 6% larger than that of HA-DEAE. Moreover, both optical micrographs and confocal laser scanning microscopy (CLSM) of the ion exchangers with adsorbed fluorescein isothiocyanate (FITC) labeled IgG revealed the superporous structure of the SA medium. In addition, the SA-DEAE column had lower backpressure than the HA-DEAE column, confirming the convective flow of mobile phase through the wide pores. Due to the presence of the wide pores, more channels were available for protein transport and, furthermore, more diffusive pores in the agarose network were accessible for the protein approach from different directions. This led to 40% higher protein capacity and two times higher effective pore diffusivity in the SA DEAE than in HA-DEAE. Moreover, an increase of the efficiency of the SA-DEAE column until a flow rate of 5 cm/min and the independency of the column efficiency at flow rates from 5 to 17.8 cm/min was found, indicating that intraparticle mass transfer was intensified by convective flow at elevated flow rates. Therefore, the chromatographic resolution of IgG and BSA was little affected up to a flow rate of 17.8 cm/min. The results indicate that the SA medium is favorable for high-speed protein chromatography. PMID- 16261632 TI - Terpenoid indole alkaloid production by Catharanthus roseus hairy roots induced by Agrobacterium tumefaciens harboring rol ABC genes. AB - We have established Catharanthus roseus hairy root cultures transgenic for the rol ABC genes from T(L)-DNA of the agropine-type Agrobacterium rhizogenes strain A4. The rol ABC hairy root lines exhibit a wild-type hairy root syndrome in terms of growth and morphology on solid medium. However, they differ from wild-type hairy root lines in that they more frequently have excellent adaptability to liquid medium and do not appear to form calli during cultivation. Moreover, they do not produce detectable levels of mannopine and agropine which, in contrast, are often synthesized abundantly in wild-type hairy root lines. The absence of these opines does not appear to cause the rol ABC lines to have higher levels of terpenoid indole alkaloids than wild-type hairy root lines. Unlike wild-type lines, rol ABC lines produce very similar levels of total alkaloids despite wide variations in individual alkaloid contents. This work demonstrates that the three genes rol ABC are sufficient to induce high-quality hairy roots in Catharanthus roseus. PMID- 16261634 TI - Tissue factor pathway inhibitor: structure, biology and involvement in disease. AB - Tissue factor (TF)-initiated coagulation plays a significant role in the pathophysiology of many diseases, including cancer and inflammation. Tissue factor pathway inhibitor (TFPI) is a plasma Kunitz-type serine protease inhibitor, which modulates initiations of coagulation induced by TF. In a factor (F) Xa-dependent feedback system, TFPI binds directly and inhibits the TF FVII/FVIIa complex. Normally, TFPI exists in plasma both as a full-length molecule and as variably carboxy-terminal truncated forms. TFPI also circulates in complex with plasma lipoproteins. The levels and the dual inhibitor effect of TFPI on FXa and TF-FVII/FVIIa complex offers insight into the mechanisms of various pathological conditions triggered by TF. The use of selective pharmacological inhibitors has become an indispensable tool in experimental haemostasis and thrombosis research. In vivo administration of recombinant TFPI (rTFPI) in an experimental animal model prevents thrombosis (and re-thrombosis after thrombolysis), reduces mortality from E. coli-induced-septic shock, prevents fibrin deposition on subendothelial human matrix and protects against disseminated intravascular coagulation (DIC). Thus, TFPI may play an important role in modulating TF-induced thrombogenesis and it may also provide a unique therapeutic approach for prophylaxis and/or treatment of various diseases. In this review, we consider structural and biochemical aspects of the TFPI molecule and detail its inhibitory mechanisms and therapeutic implications in various disease conditions. PMID- 16261633 TI - Oriented Schwann cell growth on microgrooved surfaces. AB - Silicon wafers bearing microgrooved surfaces with various groove width, spacing, and depth were fabricated using microlithography. The orientation of rat Schwann cells along the direction of the grooves was measured at 24 h after seeding the cells. When the width/spacing of the grooves was fixed at 10/10 microm, the mean percentage of aligned cells was 12% for grooves of 0.5 microm depth, 15% for those of 1 microm depth, and 26% for those of 1.5 microm depth (P < 0.05). When the depth of grooves was fixed at 1.5 microm, the mean percentage of aligned cells increased from 26% for width/spacing 10/10 microm, to 33% for 10/20 microm or 20/10 microm, and up to 41% for 20/20 microm (P < 0.05). On the surface with grooves of width/spacing/depth = 20/20/1.5 microm and modified by laminin, the alignment at 24 h approached 60%, versus 51% for collagen-coated surface and 41% for uncoated surface (P < 0.05). At 48 h after seeding, about 66% of the cells were aligned on the above laminin-modified surface. The groove depth influenced orientation of Schwann cells significantly. The cell alignment on 20/20/3 microm microgrooved poly(D,L-lactide-co-glycolide) 90:10 (PLGA) surfaces transferred from silicon reached 72% at 48 h and 92% at 72 h (P < 0.05). Coating this surface with laminin enhanced cell alignment only in short term (67% vs. 62% at 24 h, P < 0.05). The cell alignment guided by surface microgrooves was time dependent. PMID- 16261635 TI - A natural 15N approach to determine the biological fixation of atmospheric nitrogen by biological soil crusts of the Negev Desert. AB - Biological soil crusts are important cryptogamic communities covering the sand dunes of the north-western Negev. The biological crusts contain cyanobacteria and other free-living N(2)-fixing bacteria and are hence able to fix atmospheric nitrogen (N). This is why they are considered to be one of the main N input pathways into the desert ecosystem. However, up to now, in situ determinations of the N(2) fixation in the field are not known to have been carried out. We examined the natural (15)N method to determine the biological N(2) fixation by these soil crusts under field conditions. This novel natural (15)N method uses the lichen Squamarina with symbiotic green algae--which are unable to fix N(2)- as a reference in order to determine N(2) fixation. Depending on the sampling location and year, the relative biological fixation of atmospheric nitrogen was estimated at 84-91% of the total N content of the biological soil crust. The cyanobacteria-containing soil lichen Collema had a fixation rate of about 88%. These fixation rates were used to derive an absolute atmospheric N input of 10-41 kg N ha(-1) year(-1). These values are reasonable results for the fixation of atmospheric N(2) by the biological crusts and cyanolichens and are in agreement with other comparable lab investigations. As far as we are aware, the results presented are the first to have been obtained from in situ field measurements, albeit only one location of the Negev with a small number of samples was investigated. PMID- 16261636 TI - Glycosylation site analysis of human alpha-1-acid glycoprotein (AGP) by capillary liquid chromatography-electrospray mass spectrometry. AB - A new anionic surfactant (RapiGest SF) was successfully used for site-specific analysis of glycosylation in human alpha-1-acid glycoprotein (AGP). By means of this analytical approach combined with capillary HPLC-mass spectrometry (and tandem mass spectrometry), the N-linked glycosylation pattern of AGP was explored. On the basis of mass matching and MS/MS experiments ca 80 different AGP derived glycopeptides were identified. Glycosylation shows a markedly different pattern for the various glycosylation sites. At sites I and II, triantennary complex-type oligosaccharides predominate and at sites III, IV and V, tetra antennary complex-type oligosaccharides predominate. Sites IV and V show the presence of additional N-acetyl lactosamine (Gal-GlcNAc) units (even higher degree of branching and/or longer antennae are also present). PMID- 16261637 TI - How ischaemic preconditioning protects small liver grafts. AB - Interleukin-1 (IL-1) and transforming growth factor-beta (TGFbeta) are key inhibitors of hepatocyte proliferation after hepatectomy. IL-1 inhibition by heat shock proteins (HSPs) has been reported in inflammatory processes. A recent study indicated the benefits of ischaemic preconditioning in reduced-size orthotopic liver transplantation (ROLT). The present study examined: (a) the effect of ischaemic preconditioning on IL-1 and TGFbeta in ROLT; (b) whether preconditioning protects small liver grafts through HSP induction; and (c) whether the potential benefits of preconditioning on HSP is related to IL-1 inhibition. Our results, obtained with an IL-1 receptor antagonist, indicated the injurious effects of IL-1 in ischaemia-reperfusion (I/R) injury and established a relationship between IL-1 and growth factors. Thus, IL-1 reduced hepatocyte growth factor (HGF) and promoted TGFbeta release, thus contributing to the impaired liver regeneration associated with ROLT. Preconditioning inhibited IL-1 through nitric oxide (NO), thereby protecting against the injurious effects of IL 1. In addition, by another pathway independent of NO, preconditioning induced HSP70 and haem-oxygenase-1 (HO-1). HO-1 protected against I/R injury and liver regeneration, whereas the benefits resulting from HSP70 were mainly related to hepatocyte proliferation. These results suggest a mechanism that explains the effectiveness of preconditioning in ROLT. They suggest, too, that other strategies, in addition to preconditioning, that modulate IL-1 and/or HSPs could be considered in clinical situations requiring liver regeneration such as small liver grafts. PMID- 16261639 TI - Liquid chromatography/mass spectrometric analysis of rat samples for in vivo metabolism and pharmacokinetic studies of ginsenoside Rh2. AB - In vivo metabolism and pharmacokinetic studies on rat were conducted for ginsenoside Rh2, one of the components from ginseng that shows promise of anticancer activity. Liquid chromatography/mass spectrometry (LC/MS) and tandem mass spectrometry (MS/MS) with electrospray ionization were used to determine Rh2 and its metabolites in rat plasma, urine and feces. An average half-life of 16 min in plasma was obtained after intravenous administration to male Sprague Dawley rats at 5 mg/kg. No Rh2 was detected in plasma samples collected from 0 to 24 h following oral administration at 100 mg/kg, and only 0.12-0.25% of the dosed amount was found in the feces samples collected from 0 to 48 h after oral administration at 100 mg/kg. Three metabolites of Rh2 were detected in the feces samples. Oxygenation and deglycosylation were found to be the major metabolic pathways of Rh2. Intense metabolism, rather than excretion, appears to be the reason for the fast clearance of this ginsenoside. PMID- 16261640 TI - Theoretical optimization by genetic algorithm of delayed extraction parameters for a matrix-assisted laser desorption/ionization time-of-flight mass spectrometer. AB - This paper presents the application of a genetic algorithm (GA) to optimize the operating parameters, namely pulse voltage and extraction delay time, when using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOFMS). Simulations predict the presence of several combinations of these parameters that give a local maximum. The aim is to locate the optimal combination (a global maximum) of pulse voltage and extraction time delay in order to focus the ions of a particular m/z value to achieve the best resolution in a given instrumental geometry. The GA locates the global maximum quickly. The results indicate that it may be possible to achieve very high resolving power by using delayed extraction (DE)-MALDI-TOFMS with parameters obtained from the GA. PMID- 16261642 TI - Development and validation of a capillary high-performance liquid chromatography/electrospray tandem mass spectrometric method for the quantification of bisphenol A in air samples. AB - Bisphenol A (BPA) is a toxic industrial chemical that affects the endocrine system even at low concentrations. A new method, based on capillary high performance liquid chromatography/electrospray ionization tandem mass spectrometry (HPLC/ESI-MS/MS) analysis, has been developed to determine BPA in atmospheric samples. The method involves collection of air samples (typically 2 m(3)) on glass fiber filters, with ultrasonic extraction and sample concentration under vacuum before analysis. HPLC analysis was performed isocratically at a flow rate of 10 microL min(-1) using a capillary reversed-phase column and MS/MS analysis in negative ion multiple reaction monitoring (MRM) mode, using BPA-d(16) as internal standard. The present method provides linear response in the range 0.007-3.5 microg/filter (R(2) > 0.999) and is characterized by high accuracy (mean bias 2%) and good reproducibility (mean RSD 5%). High sensitivity (LOD = 2 ng/m(3) based on 2 m(3) of air collected), specificity, and speed of the analysis make the present method suitable for routine determination of BPA in the atmosphere, both for ambient and personnel monitoring. PMID- 16261641 TI - Direct infusion mass spectrometry as a fingerprint of protein-binding media used in works of art. AB - A direct infusion mass spectrometry method for the characterization of proteinaceous glues from binding media used in pictorial works of art prior to conservation or restoration treatment is proposed. Amino acids are released by acid hydrolysis and dissolved in a mixture of acidic water and ethanol. This mixture is directly infused into a mass spectrometer without any derivatization. The mass spectrometer is operated in positive ion electrospray mode (ESI-MS) to yield [M+H](+) ions for the amino acids. Relative amounts of each amino acid are calculated for each protein (beef and porcine gelatines, albumin, casein and egg). The analyzed proteins were satisfactorily distinguished. The method is easy and fast, and shows good sensitivity and resolution. The proposed method has been successfully applied to artistic samples from items of the cultural heritage of Valencia (Spain). PMID- 16261643 TI - Characterization and quantification of soy isoflavone metabolites in serum of renal transplanted patients by high-performance liquid chromatography/electrospray ionization mass spectrometry. AB - During a dietary intervention study on 16 renal transplanted patients, in which 25 g/day of animal proteins were replaced with 25 g of soy proteins, the metabolic profile of soy isoflavones in serum was characterized. This paper describes a reliable and fast liquid chromatography/electrospray ionization mass spectrometry (LC/ESI-MS) method, in negative ion mode, allowing the characterization and simultaneous quantification of several soy isoflavone metabolites. Six metabolites were identified and quantified: daidzein ([M-H](-) at m/z 252.8), dihydrodaidzein (DHD, [M-H](-) at m/z 254.8), equol ([M-H](-) at m/z 240.9), O-desmethylangolensin (O-DMA, [M-H](-) at m/z 256.8), genistein ([M H](-) at m/z 268.8), and dihydrogenistein (DHG, ([M-H(+)](-) at m/z 270.8). Quantification was assessed using two deuterated internal standards, D(3) daidzein and D(4)-genistein. This method permitted a limit of quantification (LOQ, S/N = 10) and a limit of detection (LOD, S/N = 3) of 0.05 microM and 0.005 microM for all analytes, except for genistein, where the LOQ and LOD were 0.005 microM and 0.001 microM, respectively. The linearity ranges were from 0.005 to 1.5 microM for genistein, from 0.05 to 1.5 microM for DHG, and from 0.05 to 0.7 microM for the other metabolites. The relative standard deviations (RSDs) were between 0.19% and 13.9% at the LOQ concentration for all metabolites, and between 0.6% and 4.8% at the maximum concentration. On the basis of the results obtained in the dietary intervention study, it was possible to split the patients into five groups characterized by different metabolic pathways. PMID- 16261644 TI - Application of stable isotope labeled glutathione and rapid scanning mass spectrometers in detecting and characterizing reactive metabolites. AB - The formation of reactive metabolites from a number of compounds was studied in vitro using a mixture of non-labeled and stable isotope labeled glutathione (GSH) as a trapping agent. GSH was labeled by incorporating [1,2-(13)C(2),(15)N]glycine into the tripeptide to give an overall increase of 3 Da over the naturally occurring substance. Detection and characterization of reactive metabolites was greatly facilitated by using the data-dependent scanning features of the linear ion trap mass spectrometers to give complimentary and confirmatory data in a single analytical run. A comparison was made by analyzing the samples simultaneously on a triple-stage quadrupole mass spectrometer operated in the constant neutral loss mode. The compounds studied included 2-acetamidophenol, 3 acetamidophenol, 4-acetamidophenol (acetaminophen), and flufenamic acid. GSH adducts for each of these compounds produced a characteristic pattern of 'twin ions' separated by 3 Da in the mass spectral data. This greatly facilitated the detection and characterization of any GSH-related adducts present in the microsomal extracts. Furthermore, characterization of these adducts was greatly facilitated by the rapid scanning capability of linear ion trap instruments that provided full-scan, MS/MS and MS(3) data in one single analysis. This method of detecting and characterizing reactive metabolites generated in vitro was found to be far superior to any of the existing methods previously employed in this laboratory. The combination of two techniques, stable isotope labeled glutathione and linear ion traps, provided a very sensitive and specific method of identifying compounds capable of producing reactive metabolites in a discovery setting. The complimentary set of mass spectral data (including full-scan, MS/MS and MS(3) mass spectra), obtained rapidly in a single analysis with the linear ion trap instruments, greatly accelerated identification of metabolically bioactivated soft spots on the molecules. This in turn enabled chemists to rapidly design out the potential metabolic liability from the back-up compounds by making appropriate structural modifications. PMID- 16261645 TI - Misunderstandings regarding clinical cross-over trials. PMID- 16261646 TI - Confidence interval construction for proportion difference in small-sample paired studies. AB - Paired dichotomous data may arise in clinical trials such as pre-/post-test comparison studies and equivalence trials. Reporting parameter estimates (e.g. odds ratio, rate difference and rate ratio) along with their associated confidence interval estimates becomes a necessity in many medical journals. Various asymptotic confidence interval estimators have long been developed for differences in correlated binary proportions. Nevertheless, the performance of these asymptotic methods may have poor coverage properties in small samples. In this article, we investigate several alternative confidence interval estimators for the difference between binomial proportions based on small-sample paired data. Specifically, we consider exact and approximate unconditional confidence intervals for rate difference via inverting a score test. The exact unconditional confidence interval guarantees the coverage probability, and it is recommended if strict control of coverage probability is required. However, the exact method tends to be overly conservative and computationally demanding. Our empirical results show that the approximate unconditional score confidence interval estimators based on inverting the score test demonstrate reasonably good coverage properties even in small-sample designs, and yet they are relatively easy to implement computationally. We illustrate the methods using real examples from a pain management study and a cancer study. PMID- 16261648 TI - Interactions of cationic porphyrins with double-stranded oligodeoxynucleotides: a study by electrospray ionisation mass spectrometry. AB - Electrospray ionisation mass spectrometry (ESI-MS), electrospray ionisation tandem mass spectrometry (ESI-MS/MS) and Ultraviolet-visible (UV-vis) spectroscopy were used to investigate the non-covalent interactions between small oligonucleotide duplexes with the GC motif and a group of cationic meso(N methylpyridynium-4-yl)porphyrins (four free bases with one to four positive charges, and the zinc complex of the tetracationic free base). The results obtained point to outside binding of the porphyrins, with the binding strength increasing with the number of positive charges. Fragmentations involving losses from both chains were observed for the porphyrins with N-methylpyridinium-4-yl groups in opposite meso positions. PMID- 16261649 TI - On-target trypsinization on MALDI target for MS. PMID- 16261651 TI - Stable hydrogen and oxygen isotope ratios of bottled waters of the world. AB - Bottled and packaged waters are an increasingly significant component of the human diet. These products are regulated at the regional, national, and international levels, and determining the authenticity of marketing and labeling claims represents a challenge to regulatory agencies. Here, we present a dataset of stable isotope ratios for bottled waters sampled worldwide, and consider potential applications of such data for regulatory, forensic and geochemical standardization applications. The hydrogen and oxygen isotope ratios of 234 samples of bottled water range from -147 per thousand to +15 per thousand and from -19.1 per thousand to +3.0 per thousand, respectively. These values fall within and span most of the normal range for meteoric waters, indicating that these commercially available products represent a source of waters for use as laboratory working standards in applications requiring standardization over a large range of isotope ratios. The measured values of bottled water samples cluster along the global meteoric water line, suggesting that bottled water isotope ratios preserve information about the water sources from which they were derived. Using the dataset, we demonstrate how bottled water isotope ratios provide evidence for substantial evaporative enrichment of water sources prior to bottling and for the marketing of waters derived from mountain and lowland sources under the same name. Comparison of bottled water isotope ratios with natural environmental water isotope ratios demonstrates that on average the isotopic composition of bottled water tends to be similar to the composition of naturally available local water sources, suggesting that in many cases bottled water need not be considered as an isotopically distinct component of the human diet. Our findings suggest that stable isotope ratios of bottled water have the power to distinguish ultimate (e.g., recharge) and proximal (e.g., reservoir) sources of bottled water and constitute a potential tool for use in the regulatory monitoring of water products. PMID- 16261652 TI - Matrix-assisted laser desorption/ionization mass spectrometry of taxanes. AB - Taxanes are biologically active compounds that have been extensively used in pharmacology for their powerful anticancer properties. High specificity and low level sensitivity for analysis of these compounds have been obtained with reversed-phase high-pressure liquid chromatography/mass spectrometry (RP HPLC/MS), but the number of applications of matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) for low molecular weight analytes is rapidly growing. A new MALDI-MS approach for the rapid screening of a variety of taxanes and a tandem mass spectrometric (MS/MS) analysis of the most important and diagnostic taxane fragmentation pathways are proposed. A solid phase extraction method followed by preliminary quantification is also reported. PMID- 16261653 TI - Identification of flavonoids using liquid chromatography with electrospray ionization and ion trap tandem mass spectrometry with an MS/MS library. AB - Searchable MS/MS spectra libraries, constructed using the results of liquid chromatography coupled with electrospray ionization (ESI) tandem mass spectrometry (LC/MS/MS) with data-dependent acquisition on an ion trap mass spectrometer, are presented with regard to the identification and confirmation of a variety of closely related flavonoids in a set of biological samples. Flavonoids were found to exhibit a maximum amount of structurally specific MS/MS spectra at 45% of normalized collision energy on the instrument used, without wideband activation. These MS/MS spectra were then searched automatically against a 297-substance MS/MS library that contains many previously acquired spectra of standard flavonoids. The possible applications of this powerful technique to biological samples are also discussed. Daidzein and genistein were identified through the MS/MS spectra library while searching through LC/MS/MS data for plant and microbial extracts. Moreover, these compounds proved completely distinguishable from other flavonoids of closely related structures in the MS/MS spectra library, using the NIST MS search program. The applicability of the library-searchable spectra at low concentrations was demonstrated by successful identification of daidzein and genistein at 0.05 and 0.5 microg/mL, respectively. PMID- 16261656 TI - Energy-resolved in-source collisionally induced dissociation for the evaluation of the relative stability of noncovalent complexes in the gas phase. AB - Energy-resolved in-source collisionally induced dissociation (CID) studies on the complexation of alkali metal cations by some crown ethers, nucleic acid bases, and amino acids have been performed. It has been shown that the cone voltage corresponding to the maximum ion abundance (V(c,Imax)) of the breakdown curve is characteristic of a given ion and not influenced by the cone desolvation process or the composition of the solution. Very good agreement of the V(c,Imax) value with the bond strength of the ion has been observed. Determination of the V(c,Imax) values for different ionic species is a useful, simple, and inexpensive way to obtain their relative stabilities in in-source CID conditions. PMID- 16261657 TI - Method for the elucidation of the elemental composition of low molecular mass chemicals using exact masses of product ions and neutral losses: application to environmental chemicals measured by liquid chromatography with hybrid quadrupole/time-of-flight mass spectrometry. AB - A method for elucidating the elemental compositions of low molecular weight chemicals, based primarily on mass measurements made using liquid chromatography (LC) with time-of-flight mass spectrometry (TOFMS) and quadrupole/time-of-flight mass spectrometry (LC/QTOFMS), was developed and tested for 113 chemicals of environmental interest with molecular masses up to approximately 400 Da. As the algorithm incorporating the method is not affected by differences in the instrument used, or by the ionization method and other ionization conditions, the method is useful not only for LC/TOFMS, but also for all kinds of mass spectra measured with higher accuracy and precision (uncertainties of a few mDa) employing all ionization methods and on-line separation techniques. The method involves calculating candidate compositions for intact ionized molecules (ionized forms of the sample molecule that have lost or gained no more than a proton, i.e., [M+H](+) or [M-H](-)) as well as for fragment ions and corresponding neutral losses, and eliminating those atomic compositions for the molecules that are inconsistent with the corresponding candidate compositions of fragment ions and neutral losses. Candidate compositions were calculated for the measured masses of the intact ionized molecules and of the fragment ions and corresponding neutral losses, using mass uncertainties of 2 and 5 mDa, respectively. Compositions proposed for the ionized molecule that did not correspond to the sum of the compositions of a candidate fragment ion and its corresponding neutral loss were discarded. One, 2-5, 6-10, 11-20, and >20 candidate compositions were found for 65%, 39%, 1%, 1%, and 0%, respectively, for the 124 ionized molecules formed from the 113 chemicals tested (both positive and negative ions were obtained from 11 of the chemicals). However, no candidate composition was found for 2% of the test cases (i.e., 3 chemicals), for each of which the measured mass of one of the product ions was in error by 5-6.7 mDa. PMID- 16261658 TI - Regulation and interplay of apoptotic and non-apoptotic cell death. AB - Various death triggers including DNA damage, oxidative stress, and growth factor deprivation promote the loss of mitochondrial membrane potential, leading to the production of reactive oxidative species (ROS) or enhanced permeability of the mitochondrial membrane, otherwise known as mitochondrial membrane permeabilization, by insertion of Bax/Bak into the outer membrane where it interacts with voltage-dependent anion channel (VDAC)/adenine nucleotide transporter (ANT). MMP leads to the release of small pro-apoptotic molecules, which induce caspase-dependent and -independent apoptotic cell death. The production of ROS due to the loss of mitochondrial membrane potential enhances the permeability of lysosomal membranes, resulting in the release of lysosomal proteases, which contribute to mitochondrial membrane permeabilization and the lysosomal degradation mechanism of autophagic cell death. Although defects in apoptotic and non-apoptotic cell death pathways can be carcinogenic, these pathways are more or less preserved within cancer cells and can therefore influence cell death and mediate resistance to cancer treatment. This paper discusses recent advances in determining the molecular mechanisms behind regulation of apoptotic and non-apoptotic cell death, as well as the interplay between these two processes, which may lead to the development of new strategies by which to enhance the therapeutic effects of chemotherapeutic agents. PMID- 16261660 TI - (1)H and (19)F PGSE diffusion and HOESY NMR studies on cationic palladium (II) 1,3-diphenylallyl complexes in THF solution. AB - THF solutions of the cationic chiral 1,3-diphenylallyl bidentate phosphine complexes [Pd(eta(3)-PhCHCHCHPh)(Duphos)](CF(3)SO(3)), Duphos = 1,2-Bis-((2R,5R) 2,5-dimethylphospholano)benzene), 2, and [Pd(eta(3)-PhCHCHCHPh)(P,S)]BF(4), 4, P,S = [8-((o-(diphenylphosphino)benzyl) thiomethyl]-(7,7'-dimethyl)-exo norborneol, have been studied via pulsed gradient spin-echo (PGSE) diffusion, (1)H, (19)F HOESY and a variety of other multi-dimensional NMR methods. On the basis of the (1)H, (19)F HOESY data, the anions show a preference for a specific structural position with respect to the eta(3)-PhCHCHCHPh allyl ligand, i.e. the anion does not move evenly around the periphery of the cation. THF is shown to promote significant ion pairing, although neither 2 nor 4 shows 100% ion pairing. PMID- 16261661 TI - Cardiovascular complications in patients with advanced prostatic cancer treated by means of orchiectomy or polyestradiol phosphate. AB - OBJECTIVE: To evaluate the cardiovascular (CV) complications associated with orchiectomy (OE) and parenteral polyestradiol phosphate (PEP) therapy (240 mg/month), taking into account the effect of pretreatment diseases and pretreatment medication. MATERIAL AND METHODS: A total of 244 T3-4 M0 patients and 200 T1-4 M1 patients were randomized to either OE or PEP therapy. The two groups of patients were analyzed separately. The follow-up period was 36 months. The effect of pretreatment vascular and other diseases and pretreatment medication which may be associated with a risk of CV complications was evaluated. RESULTS: In the T3-4 M0 patients, the treatment (PEP versus OE) and the presence of pretreatment vascular diseases were statistically significantly associated with a risk of CV complications (p=0.01 and 0.003, respectively). In the T1-4 M1 patients, such an association was not found. No association was observed between pretreatment medication and CV complications. There was no difference in progression-free time between the therapy groups in either the T3-4 M0 or T1-4 M1 patients. CONCLUSION: In patients with locally advanced prostatic cancer, PEP therapy is associated with a statistically significantly higher risk of CV complications compared to OE. PMID- 16261662 TI - Obstructive sleep apnoea and plasma homocysteine. PMID- 16261663 TI - [Diagnostic procedures in the suspected presence of a blood cell dyscrasia]. PMID- 16261664 TI - ESPEN guidelines on artificial enteral nutrition--percutaneous endoscopic gastrostomy (PEG). PMID- 16261665 TI - Supervisor-trainee relationship boundaries in medical education. AB - Despite concerns about the prevalence and ramifications of medical student mistreatment, the boundaries of faculty-student relationships have not been studied systematically in medical training programs. This study aimed to identify behaviours that occur with some frequency and potentially raise issues related to boundaries in the supervisor-trainee relationship. An anonymous questionnaire was distributed to the mailboxes of 154 residents in the departments of psychiatry, internal medicine, and obstetrics and gynaecology at four hospitals affiliated with Brown University Medical School. Residents were asked to report whether they had encountered specific behaviours from supervisors during medical training, the frequency of these experiences, and the professional status of the supervisor involved in each episode. There was a significant reported incidence of behaviours related to academic/professional boundaries, personal boundaries, and dating boundaries. Some of these behaviours raise issues related to exploitation. The major sources of these reported boundary behaviours were hospital-based clinical faculty, university-based academic faculty, and senior house staff. The potentially adverse effects of boundary behaviours on the individual student, the teacher-student relationship, and the doctor-patient relationship are discussed. Future research is recommended to clarify the limits of appropriate behaviour between supervisors and trainees in the medical learning environment PMID- 16261666 TI - Applying the case method for teaching within the health professions--teaching the teachers. AB - The case method provides training in problem solving and is useful in clinical education within health professions. The method consumes less teaching resources and might be useful in a situation with increasing numbers of students and staff cuts. As it is a relatively new method within the Faculty of Medicine, University of Lund, a training program for teachers has been established with a short, practical, course. This paper describes the application and introduction of the case method and evaluation of the introductory course by 106 teachers. In this course the case method is introduced and demonstrated. The participants construct cases and take an active part as both students and teachers. Almost everyone was planning to incorporate the case method in their teaching practice, at both undergraduate and graduate levels. To have constructed and led cases themselves was considered a great strength. More time was requested as well as a follow-up session after practically using the case method. PMID- 16261667 TI - Perspectives on management education: an exploratory study of UK and Portuguese medical students. AB - Healthcare management is becoming extremely important and large health organizations face increasing demands for leadership and system change. The role of doctors is pivotal but their relationship with management issues and practice has been a matter of long-lasting debate. The aim of this research was to establish opinions of medical students and other medical educational stakeholders on the value and structure of a management and leadership course in medical school. A survey of undergraduate medical students from two medical schools (n = 268) was carried out, and quantitative and qualitative data were analysed and compared with opinions collected from interviews with hospital managers and clinical professors. Portuguese medical students attributed higher relevance to leadership/management education than their UK counterparts. For both groups, such a course would be best: (1) situated in the clinical years, (2) optional and (3) one term/semester long. Main topics desired were 'Managing people/team management'; 'National Health Service'; 'Doctors and Leadership', 'Costs/prices and resource management'. In conclusion, leadership/management education is perceived as relevant but its inclusion in the medical curriculum as well as its content needs careful consideration. Education in informatics and knowledge management would also provide a positive contribution to professional development but is scarcely appreciated at present. PMID- 16261668 TI - A pilot survey of junior doctors' confidence in tasks related to broad aspects of competence. AB - Medical authorities and society are recommending that postgraduate medical education prepare physicians for broad aspects of competence. The most effective ways to prepare physicians for these are not known. The aim of this pilot study was to survey junior doctors' growth in confidence in different physician roles. A total of 165 junior doctors in internal medicine completed a self-administered survey of confidence levels in physician roles. Confidence levels between training levels were compared. Confidence in the roles of medical expert, communicator and collaborator increased between PRHOs and SHOs. Confidence levels increased in all roles between PRHOs and SpRs, but confidence between SHOs and SpRs increased only in the roles of collaborator and scholar. Confidence in the role of professional was low and did not increase significantly between training levels. The SHO year was rated as only moderately useful in facilitating learning of the tasks. These results suggest that growth of confidence in the physician roles proceeds at different rates during postgraduate training. Additional research is needed to identify effective ways for junior doctors to learn broad aspects of competence. Longitudinal tracking will be important to evaluate the effect of educational interventions on patient care. PMID- 16261669 TI - Learning in a web-based system in medical education. AB - New learning environments such as distance education and computer-aided instruction promise to bring a change in today's learning environments by adjusting the relationship between the learner, the educational content and the organization of education. In this study, we explored whether students' approaches to learning related to their perception of a particular virtual learning environment. Scales of the ASSIST questionnaire were loaded in a two principal component solution, surface and deep-strategic. We found statistically significant correlations between the approaches to learning and the student's attitudes towards ICT. Early identification of approaches to learning and attitudes towards ICT may prove to be important in order to provide assistance to aid the transition of students with diverse individual characteristics and to the design of new learning environments. PMID- 16261670 TI - A longitudinal curriculum to improve resident teaching skills. AB - Residents are essential teachers of students and interns, yet few opportunities exist for them to improve their teaching skills. The authors designed and implemented a longitudinal teaching skills curriculum for second-year categorical medicine residents on ambulatory block. This curriculum involves residents giving talks to one another, with close observation and personalized guidance by faculty in one-on-one and group feedback sessions. Evaluation by self-assessment surveys administered at the beginning and end of the curriculum showed significant improvement in nearly all of the domains assessed, including overall teaching skills. Intensive interventions can improve resident teaching proficiency and thereby improve the education provided to trainees. PMID- 16261671 TI - Romanians pay 1 million dollars extra a day for health. PMID- 16261672 TI - Mental health and religion in Israel's ultra-Orthodox Jews. PMID- 16261674 TI - Effects of repeated human chorionic gonadotropin administration on serum testosterone and testicular volume in prepubertal Thoroughbred colts. PMID- 16261673 TI - Belgian health minister targets mental health "charlatans". PMID- 16261675 TI - Genetic analysis of the fertility in Hanoverian Warmblood horses. PMID- 16261677 TI - The effect of sperm number and frequency of insemination on pregnancy rates of mares inseminated with frozen-thawed spermatozoa. PMID- 16261676 TI - Influence of the polyol pathway on norepinephrine transporter reduction in diabetic cardiac sympathetic nerves: implications for heterogeneous accumulation of MIBG. AB - PURPOSE: Cardiac scintigraphic studies using 123I-labeled metaiodobenzylguanidine ([123I]MIBG) have demonstrated heterogeneous myocardial accumulation of MIBG in diabetes. The accumulation has been found to correlate with a heterogeneous decrease in the expression of norepinephrine transporter (NET). In diabetic peripheral nerve tissue, polyol pathways are activated and cause nerve dysfunction and degeneration. However, there has been little research on the polyol pathway and cardiac sympathetic nerves. Therefore, to assess the influence of the polyol pathway on cardiac sympathetic nervous function, we investigated the regional accumulation of MIBG and NET protein expression in diabetic model rats treated with aldose reductase inhibitor (ARI) for the blockade of polyol pathways. METHODS: Rats were given a single intravenous injection of streptozotocin (n=76, STZ-D rats). Starting the day after STZ injection, ARI was administered daily to 42 of the rats for 4 weeks (ARI-D rats). To assess the cardiac sympathetic nervous function, [125I]MIBG autoradiographic experiments were carried out. Finally, NET protein expression was assessed with a saturation binding assay. RESULTS: The myocardial sorbitol concentration was significantly higher in STZ-D rats than in ARI-D rats. There was no heterogeneous accumulation of MIBG in ARI-D rats. There was a heterogeneous decrease of NET expression in STZ-D rats, but not in ARI-D or control rats. CONCLUSION: The gathered data indicate that the enhanced polyol pathway correlates with the decrease in regional cardiac sympathetic nervous function, and this impairment may lead to the reduction of NET protein in cardiac sympathetic nerves of the diabetic inferior wall. PMID- 16261678 TI - Influence of cryopreserved sperm or semen extender on uterine contractile activity in mares following conventional or hysteroscopic insemination. PMID- 16261679 TI - Effects of frequency of insemination, number of spermatozoa and insemination site on fertility of equine frozen semen. PMID- 16261680 TI - Selenium contents in equine semen and semen fractions and their relations with chromatin integrity and foal birthing rate. PMID- 16261681 TI - Evaluation and diagnosis of acrosome function/dysfunction in the stallion. PMID- 16261682 TI - Cholesterol to phospholipid ratio in sperm of stallions with unexplained subfertility. PMID- 16261683 TI - [The limits of empathy]. PMID- 16261684 TI - Hypoosmotic swelling test: a new approach by multi-hypotonic steps. PMID- 16261685 TI - For the patient. Race, stress and the health of African-American women. PMID- 16261686 TI - For the patient. Pulse pressure among US adults. PMID- 16261687 TI - For the patient. Diabetes affects your heart, too. PMID- 16261688 TI - For the patient. How fat tissue affects disease. PMID- 16261689 TI - For the patient. Hair relaxer not linked to early births. PMID- 16261690 TI - For the patient. Genetic science working to find solutions for healthier hearts. PMID- 16261692 TI - A simple method for characterising syndactyly in clinical practice. AB - Non-syndromic syndactyly is a heterogeneous group of limb malformations involving webbing of fingers and/or toes. There are at least nine non-syndromic types described in the literature. For the clinician and the genetic counsellor not having gathered experience with this malformation, it is rather tedious to identify the correct subtype for the patient's phenotype. We therefore present a protocol for clinical use, which visualises the malformation in a graphical way and thereby simplifies typing. In addition, this protocol provides a simple documentation system for reporting clinical data for new syndactyly families. It might encourage clinicians to report families that are still unclassifed and thus, helping to extend and improve the existing classification system. PMID- 16261691 TI - For the patient. Exercise and how it affects risk factors for heart disease among Asian Indians. PMID- 16261693 TI - Zimmermann-Laband syndrome: further clinical delineation. AB - Zimmermann-Laband syndrome (ZLS) is an autosomal dominant disorder characterized by gingival fibromatosis, absent or dysplastic distal phalanges, vertebral defects, hepatosplenomegaly, hypertrichosis and sometimes mental retardation. We describe two unrelated patients, a girl aged 9 years and a boy 11 months whose clinical and radiological findings permit us to diagnose the ZLS. Body overgrowth, present in both patients, was identified as a main clinical feature not previously reported as well as the presence in neuroimaging studies of a cavernous hemangioma on the frontal and the left cerebellar regions in the boy. The girl also presented important radiological characteristics such as broad medulary canals and metaphyses of long bones, thin cortices, broad ribs, accelerated skeletal maturation as well as high intelligence level. A wide clinical spectrum in ZLS is also considered. PMID- 16261694 TI - Metastatic adenocarcinoma of right supraclavicular fossa--delayed presentation of ovarian primary. PMID- 16261695 TI - Item method directed forgetting diminishes false memory. AB - Directed forgetting may reduce DRM false memory illusion by interfering with meaning processing. Participants were presented with a list composed of six 10 word semantically associated sub-lists, and they were either (a) asked to remember all list items of (b) asked to remember all associates from sub-lists and to forget all associates from other sub-lists. All participants were requested to recall and recognise list items. Although directed forgetting effects have been previously reported only for true responses in the DRM paradigm with the list method, we also found directed forgetting for false responses with the item method. Such forgetting instructions reduced both verbatim and meaning processing, decreasing both the intrusion and the false alarm rate. These results are consistent with two-process explanations of DRM false memories, such as fuzzy trace theory, and add to our understanding of false memory editing. PMID- 16261696 TI - [Ambulatory chemotherapy in the Kitasato University Hospital]. PMID- 16261697 TI - [Management of ambulatory chemotherapy in a hospital]. PMID- 16261698 TI - Abstracts from the 30th Congress of the Society of Biomechanics, Brussels, Belgium, 14-16 September 2005. PMID- 16261699 TI - The official 2005-2006 directory of members of the Texas Dental Association. PMID- 16261700 TI - Abstracts of the 37th Annual General Meeting of the European Brain and Behaviour Society, Dublin, Ireland, September 24-28, 2005. PMID- 16261701 TI - Abstracts of the 18th World Congress on Psychosomatic Medicine, August 21-26, 2005, Kobe, Japan. PMID- 16261702 TI - CHEST 2005. Abstracts. October 29-November 3, 2005, Montreal, Canada. PMID- 16261703 TI - Methotrexate" "lessons from rheumatology". PMID- 16261704 TI - Anticoagulants and gastrointestinal bleeding. PMID- 16261705 TI - Characteristics of patients referred to Alcoholism Treatment Centers. AB - Data collected in a pilot study of 57 Alcoholism Treatment Centers were used to test the hypothesis that different racial, age, and sex groups were associated with different referral sources. Regression analysis was used to predict types of referral from specific demographic groups. The analysis confirmed the hypothesis, suggesting a possible discriminatory referral process. PMID- 16261706 TI - Limited take-up of health coverage tax credits: a challenge to future tax credit design. AB - The Trade Act of 2002 created federal tax credits to subsidize health coverage for certain early retirees and workers displaced by international trade. Though small, this program offers the opportunity to learn how to design future tax credits for larger groups of uninsured. During September 2004, the most recent month for which there are data about all forms of Trade Act credits, roughly 22 percent of eligible individuals received credits. The authors find that health insurance tax credits are more likely to reach their target populations if such credits: 1) limit premium costs for the low-income uninsured and do not require full premium payments while applications are pending; 2) provide access to coverage that beneficiaries value, including care for preexisting conditions; 3) are combined with outreach that uses easily understandable, multilingual materials and proactive enrollment efforts; and 4) feature a simple application process involving one form filed with one agency. PMID- 16261707 TI - [Pharmacogenomics]. PMID- 16261708 TI - [Tropical or non-tropical sprue?]. AB - A 57-year-old Dutch man presented with weight loss and fatigue 6 months after a visit to West Papua, when he had suffered from serious diarrhoea. Macrocytic anaemia and vitamin B12 deficiency were diagnosed. A gastroduodenoscopy with biopsies of the small intestine was performed revealing no macroscopic abnormalities but partial villous atrophy was found microscopically, suggesting tropical sprue or coeliac disease. Antibodies against endomysium and tissue transglutaminase were negative, ruling out coeliac disease. The patient was successfully treated with vitamin B12, folic acid and doxycycline. This case shows that tropical sprue should be considered in the differential diagnosis of chronic diarrhoea in patients with a history of travel in tropical regions. The most frequent medical problem that travelers to the tropics experience is diarrhoea with an incidence of 30%. A small proportion of these patients eventually present with chronic diarrhoea. At that moment, the relation to their previous travelling may not be immediately clear. One of the causes of this chronic diarrhoea to be considered is tropical sprue. PMID- 16261709 TI - [Stroke services are increasingly effective]. AB - Since the first Consensus Statement on the treatment of stroke (UK, 1988) and the European Helsingborg Declaration on the treatment of stroke (1995), 69 stroke services have been geographically spread in the Netherlands; these consist of an integrated hospital stroke unit, a nursing home rehabilitation unit, and home care. The Dutch stroke services have improved their outcomes in a cost-effective way in comparison with the usual care. Consensus on patient-oriented outcomes- arranged in order of the International Classification of Functioning, Disability and Health--is recommended. PMID- 16261710 TI - [Radio-immunotherapy for B-cell non-Hodgkin lymphoma]. AB - Radio-immunotherapy is a new treatment modality for patients with B-cell non Hodgkin lymphoma. In radio-immunotherapy, a therapeutic radionuclide is coupled to a monoclonal antibody directed against a tumour-specific or tumour-associated antigen. Biodistribution studies and dosimetry are used in the planning of radio immunotherapy. Clinical studies, notably in patients with indolent non-Hodgkin lymphoma, have demonstrated the clinical feasibility and efficacy of this treatment. The use of a high dose ofradio-immunotherapy in combination with (high dose) chemotherapy and peripheral stem-cell transplantation constitutes a supplemental treatment for patients who respond insufficiently or not at all to standard therapy. The exact place of radio-immunotherapy in the treatment of patients with non-Hodgkin lymphomas is not yet clear. PMID- 16261711 TI - [Prevention of contrast nephropathy; guidelines from the Department Nephrology of the University Medical Centre Groningen]. AB - Contrast nephropathy' is commonly defined as an increase in serum creatinine concentration of more than 44 micromol/l or 25% of the reference value within 48 hours after administration of radiological contrast media. Risk factors are pre existing renal dysfunction, diabetes mellitus, heart failure, paraproteinaemia, advanced age, use of nephrotoxic medication, intra-arterial use of contrast media and use of large volumes of contrast media. The only form of therapy possible is supportive care, such as renal function replacement therapy. Prevention of contrast nephropathy is of great importance. For patients with risk factors, the treatment of first choice is hydration with intravenous sodium chloride 0.9% combined with oral acetylcysteine given before and after the administration of contrast media. PMID- 16261712 TI - [From gene to disease; Leber congenital amaurosis (LCA)]. AB - LCA is a severe retinal dystrophy characterised by an onset of symptoms before the age of 6 months, visual acuity below 201/400, searching nystagmus, sluggish pupillary reactions and no detectable responses on electrography. The visual fields are usually not measurable. LCA is genetically heterogeneous and is usually inherited in an autosomal recessive fashion. Seven genes have been reported to be mutated in LCA patients (AIPL1, CRB1, CRX, GUCY2D, RDH12, RPE65 and RPGRIP1). Each gene is responsible for a fraction of LCA patients. Mutations in these seven genes are estimated to underlie approximately 40-50% of LCA cases. Molecular genetic research is crucial to unravel the remaining genetic causes of this disabling disease. PMID- 16261713 TI - [Diagnostic image (249). An infant with a swollen red toe]. AB - A 4-month-old male infant had a red and swollen left fourth toe due to strangulation by a hair. PMID- 16261714 TI - [Participation in the Dutch national screening programme for uterine cervic cancer higher after invitation by a general practitioner, especially in groups with a traditional low level of attendance]. AB - OBJECTIVE: To gain insight into the differences in participation in the screening programme for uterine cervix cancer between women invited by a general practitioner (GP) and women invited by the local health authority (GGD). Specific attention was given to those groups whose participation is generally below average. DESIGN: Descriptive epidemiological study. METHOD: In the period 2000 2003, 237,719 women (30-60 years of age) were invited to participate in the national uterine cervix cancer screening programme in the Southwest of the Netherlands; 37.1% of the women were invited by the GP and 62.9% by the GGD. Data were obtained from the Cervix Information System of the GGDs. Differences in attendance between those invited by the GP and those invited by the GGD were tested by linear regression. Participation was defined as the number of women for whom the result of a cervical smear was known, divided by the number invited. RESULTS: Invitation by a GP led to a 7.9% (95% CI: 7.5-8.3) higher attendance rate than invitation by a GGD. This difference in attendance was higher for women born in Morocco, Turkey, Surinam and the Netherlands Antilles/Aruba (17.2%; 95% CI: 15.2-I9.2), young women (11.9%; 95% CI: 10.8-13.0), women with a low socio economic status (11.6%; 95% CI: 10.4-12.7), and women who lived in highly urban areas (13.0%; 95% CI: 12.3-13.6). The differences were the greatest among non western women who were also part of another low-attendance group: 19.0% (95% CI: 16.7-21.2) for women who lived in highly urban areas and 20.8% (95% CI: 16.8 24.9) for those in the youngest age group. PMID- 16261715 TI - [Efficacy of agreements within the Enchede Stroke Service to refer patients with a stroke from the stroke unit in the hospital to a nursing home for short-term rehabilitation]. AB - OBJECTIVE: To assess the efficacy of agreements within the Enschede Stroke Service to refer patients with a stroke from the stroke unit in the hospital to a nursing home for short-term rehabilitation. DESIGN: Prospective, partly retrospective. METHOD: All patients who were referred from the stroke unit at Medisch Spectrum Twente to the CVA Rehabilitation Unit (CRU) in the period 1 July 1999-31 July 2003 were included. Referral took place via an active multidisciplinary approach and specific referral agreements. The primary outcome was the number of patients that could be discharged home after rehabilitation. In addition, we assessed the influence on final discharge destination of age, the Barthel and Rankin scores at the time of admission to the CRU and the medical complications during the period of rehabilitation. RESULTS: 232 patients were included (133 women and 99 men, mean age 76.4 years). Within 3 months, 63% of the patients were discharged home. After 6 months, 82% had returned home. 8% of the patients died within 6 months and 9% had to stay in a nursing home permanently. Of the patient aged 80 years or older, 75% could return home within 6 months. Patients with poor Barthel and Rankin scores and medical complications had a smaller chance of being discharged home. CONCLUSION: Effective referral of patients from the stroke unit to a nursing home for short-term rehabilitation is possible. With adequate patient selection, the use of good referral agreements and multidisciplinary consultations, most patients could finally return home. PMID- 16261716 TI - [Rituximab instead of splenectomy in 4 children with chronic or refractory autoimmune haemolytic anaemia]. AB - 4 children, a boy aged 10 years and 3 girls aged 3, 3, and 16 years, suffering from chronic or refractory autoimmune haemolytic anaemia (AIHA), who were dependent on high doses of steroids and were refractory to immunosuppressants, were treated with rituximab at a dose of 375 mg/m2 once a week for 3 or 4 weeks as an alternative to splenectomy. Rituximab is a monoclonal anti-CD20 antibody that prevents the production ofautoantibodies by selective destruction of B lymphocytes. Haemoglobin levels increased and the parameters of chronic haemolysis (reticulocyte count, lactate dehydrogenase activity, bilirubin concentration) decreased to normal values. 3 patients were taken off corticosteroids completely; 1 of these was also no longer dependent on blood transfusions. Circulating B-lymphocytes were absent for 6 to 15 months after the treatment and the rituximab was well-tolerated. During the treatment, immunoglobulins were substituted and infectious complications were not seen. Rituximab was valuable in the treatment of chronic or refractory AIHA and eliminated the need for splenectomy. 1 patient did not respond to rituximab. PMID- 16261717 TI - [The influenza season 2004/'05 in the Netherlands with the largest epidemic of the last 5 years caused by the virus variant A/California and the composition of the vaccine for the season 2005/'06]. AB - In the Netherlands, the influenza epidemic of the 2004/'05 season started late. The background value of 3 cases of an influenza-like illness per 10,000 inhabitants per week was exceeded from week 1 until week 14 of 2005. The magnitude of the epidemic was the largest of the last 5 years, namely 104 per 10,000 inhabitants. As usual, the epidemic was caused mainly by influenza-A viruses of subtype H3N2 and to a lesser degree by A/H1N1 and B viruses. The H3N2 virus isolates belonged to the newly emerged variant A/California/7/04, which deviated slightly from the vaccine strain used for the 2004/'05 season. The influenza-B and H1N1 viruses matched the corresponding vaccine viruses well. For the 2005/'06 season, the World Health Organization has recommended the following vaccine composition: A/California/7/04 (H3N2), A/New Caledonia/20/99 (HiNI), and B/Shanghai/361/02. PMID- 16261718 TI - [Medical specialist training at the College of Medicine in Malawi; the value of the Dutch contribution]. AB - Malawi, one of the world's poorest nations, has been until recently largely dependent on foreign doctors. In 1991 the College of Medicine was founded in Blantyre, Malawi, to train doctors locally, using a curriculum that meets international standards and is tailored to local needs. The Dutch government has supported this initiative financially and by providing medical specialists to help develop the curriculum, to teach and to assist in clinical and research tasks. The College has been remarkably successful. Most graduates remain to practice in Malawi and student numbers have increased from 30 to 65 per year. A training programme for medical specialists has been recently started that is aimed at providing university staff who can take over from the expatriates. It will still take several years before the College is able to train sufficient, qualified local teaching staff. Continued support from the Dutch government is essential as well as intensified cooperation with Dutch academic centres in medical education, specialist training and research. PMID- 16261719 TI - [The guideline 'Diagnosis and treatment of peripheral artery disease of the lower extremities' of The Netherlands Surgical Society]. PMID- 16261721 TI - [Cannabinoid hyperemesis with the unusual symptom of compulsive bathing]. PMID- 16261722 TI - [Use of alcohol during conception, pregnancy and lactation]. PMID- 16261723 TI - [Metabolic syndrome]. PMID- 16261724 TI - Higher disease prevalence can induce greater sociality: a game theoretic coevolutionary model. AB - There is growing evidence that communicable diseases constitute a strong selective force on the evolution of social systems. It has been suggested that infectious diseases may determine upper limits of host sociality by, for example, inducing territoriality or early juvenile dispersal. Here we use game theory to model the evolution of host sociality in the context of communicable diseases. Our model is then augmented with the evolution of virulence to determine coevolutionarily stable strategies of host sociality and pathogen virulence. In contrast to a controversial hypothesis by Ewald (1994), our analysis indicates that pathogens may become more virulent when contact rates are low, and their prevalence can ultimately induce greater sociality. PMID- 16261725 TI - The effect of self-fertilization, inbreeding depression, and population size on autopolyploid establishment. AB - The minority cytotype exclusion principle describes how random mating between diploid and autotetraploid cytotypes hinders establishment of the rare cytotype. We present deterministic and stochastic models to ascertain how selfing, inbreeding depression, unreduced gamete production, and finite population size affect minority cytotype exclusion and the establishment of autotetraploids. Results demonstrate that higher selfing rates and lower inbreeding depression in autotetraploids facilitate establishment of autotetraploid populations. Stochastic effects due to finite population size increase the probability of polyploid establishment and decrease the mean time to tetraploid fixation. Our results extend the minority cytotype exclusion principle to include important features of plant reproduction and demonstrate that variation in mating system parameters significantly influences the conditions necessary for polyploid establishment. PMID- 16261726 TI - Sexual selection when fertilization is not guaranteed. AB - Much of the theory of sexual selection assumes that females do not generally experience difficulties getting their eggs fertilized, yet sperm limitation is occasionally documented. How often does male limitation form a selection for female traits that improve their mating rate? The question is difficult to test, because if such traits evolve to be efficient, sperm limitation will no longer appear to be a problem to females. Here, we suggest that changes in choosiness between populations, and in particular between virgin and mated females, offer an efficient way to test this hypothesis. We model the "wallflower effect," that is, changes in female preferences due to time and mortality costs of remaining unmated (for at least some time). We show that these costs cause adaptive reductions of female choice, even if mate encounter rates appear high and females only rarely end their lives unfertilized. We also consider the population consequences of plastic or fixed mate preferences at different mate encounter rates. If mate choice is plastic, we confirm earlier verbal models that virgins should mate relatively indiscriminately, but plastic increase of choosiness in later matings can compensate and intensify sexual selection on the male trait, particularly if there is last male sperm precedence. Plastic populations will cope well with unusual conditions: eagerness of virgins leads to high reproductive output and a relaxation of sexual selection at low population densities. If females lack such plasticity, however, population-wide reproductive output may be severely reduced, whereas sexual selection on male traits remains strong. PMID- 16261727 TI - Cryptic speciation in the cosmopolitan and clonal human pathogenic fungus Aspergillus fumigatus. AB - Microbes and other organisms smaller than one to a few millimeters in size are hypothesized to have global populations, in contrast to the geographically restricted ranges of larger organisms. However, fungi, which routinely have reproductive propagules no larger than 10 micrometers, challenge the generality of this hypothesis because recent studies have shown that globally distributed morphological species embrace two or more geographically restricted phylogenetic species. We used the concordance of gene genealogies to recognize phylogenetic species in the globally distributed opportunistic human pathogenic fungus, Aspergillus fumigatus. Based on DNA sequence data of five loci for each of 63 individuals collected from five continents, we have delineated two phylogenetic species in this single morphological species. Unlike all other fungi examined to date, both genetically isolated groups showed a global distribution with no evidence of a correlation between genotype and geographic location. Sexual reproduction has never been observed in A. fumigatus, but when the same data were used to explore the association of alleles at the five loci for one of the phylogenetic species, evidence was found to support recombination. The discovery of a cryptic species is medically relevant because different species are likely to differ in virulence or drug resistance. The discovery of a globally distributed A. fumigatus species clade highlights the need for ecological studies of the fungus to either document global dispersal or propose alternative mechanisms by which it persists as single, global phylogenetic population. PMID- 16261728 TI - Interaction between founder effect and selection during biological invasion in an aquatic plant. AB - Long-distance colonization and rapid range expansion associated with biological invasion may have major evolutionary consequences via both stochastic processes and selection. Using large-scale population genetic surveys, we demonstrate a major shift in the relative frequency of sexually fertile diploid versus sexually sterile triploid populations associated with the invasion of North America by a clonal aquatic plant, Butomus umbellatus. Most populations across the native European range were triploid (84% of 108), whereas most introduced populations were diploid (71% of 136). We evaluated the roles of stochastic processes versus natural selection in causing this shift by surveying predominantly neutral genetic variation at 28 RAPD loci. In Europe (EU) we detected 47 distinct genotypes among 142 plants sampled from 71 populations, whereas in North America (NA) we detected only six genotypes among 138 plants from 69 populations. Of the six NA genotypes, a set of four closely related genotypes were found only in triploid populations and a pair of closely related genotypes were found only in diploid populations, and these were genetically divergent from the triploid genotypes. This result is consistent with severe founder effect. Because sex creates genotypic variation and produces offspring with greater dispersal potential than those produced clonally, we tested the hypothesis that sexual reproduction characteristic of diploids has given them a colonization advantage that accounts for their high frequency in NA. However, we found little or no evidence of sexual recruitment in introduced diploids. One very widespread heterozygous genotype occurred in 95% of 38 introduced diploid populations (i.e., 72 of 76 plants surveyed) suggesting predominant clonal reproduction. Moreover genotypic diversity was not higher within or among diploid than triploid populations in either the native or introduced range. Low genetic diversity in diploid populations was also supported by a comparison of within-population quantitative variation for plant size under a common greenhouse environment. Thus, diploids have not been favored during colonization owing to their sexual fertility. However, concurrent studies have shown that NA diploids exhibit a much higher capacity for clonal reproduction, via small vegetative bulbils, than NA triploids, which almost never produce bulbils. The same difference in clonal capacity is not a consistent feature of the native EU populations. Taken together, these results suggest that strong founder effect has set the stage for a major increase in diploid frequency due to the particular, and possibly idiosyncratic, features of the diploid and triploid lineages introduced to North America. PMID- 16261729 TI - Climate and life-history evolution in evening primroses (Oenothera, Onagraceae): a phylogenetic comparative analysis. AB - Evolutionary ecologists have long sought to understand the conditions under which perennial (iteroparous) versus annual (semelparous) plant life histories are favored. We evaluated the idea that aridity and variation in the length of droughts should favor the evolution of an annual life history, both by decreasing adult survival and by increasing the potential for high seedling survival via reduced plant cover. We calculated phylogenetically independent contrasts of climate with respect to life history in a clade of winter-establishing evening primroses (sections Anogra and Kleinia; Oenothera; Onagraceae), which includes seven annuals, 12 perennials, and two variable taxa. Climate variables were quantified from long-term records at weather stations near collection localities. To explicitly account for phylogenetic uncertainty, contrasts were calculated on a random sample of phylogenetic trees from the posterior distribution of a Bayesian analysis of DNA sequence data. Statements of association are based on comparing the per-tree mean contrast, which has a null expectation of zero, to a set of per-tree mean contrasts calculated on the same trees, after randomizing the climate data. As predicted, increased annual aridity, increased annual potential evapotranspiration, and decreased annual precipitation were associated with transitions to the annual habit, but these trends were not significantly different from the null pattern. Transitions to the annual habit were not significantly associated with increases in one measure of aridity in summer nor with increased summer drought, but they were associated with significantly increased maximum summer temperatures. In winter, increased aridity and decreased precipitation were significantly associated with transitions to the annual habit. Changes in life history were not significantly associated with changes in the coefficient of variation of precipitation, either on an annual or seasonal (summer vs. winter) basis. Though we cannot attribute causality on the basis of a correlational, historical study, our results are consistent with the idea that increased heat and drought at certain times of the year favor the evolution of the annual habit. Increased heat in summer may cause adult survival to decline, while increased aridity and decreased precipitation in the season of seedling recruitment (winter) may favor a drought-avoiding, short-lived annual strategy. Not all of the predicted patterns were observed: the capability for drought induced dormancy may preclude change in habit in response to summer drought in our study group. PMID- 16261730 TI - Selection for phenotypic divergence between diploid and autotetraploid Heuchera grossulariifolia. AB - Much of the diversity of flowering plants is associated with genomic duplication through polyploidy. Little is known, however, about the evolutionary mechanisms responsible for the diversification of novel polyploid lineages. We evaluated the possibility that divergence is driven by natural selection by estimating the strength of phenotypic selection acting on three floral traits in sympatric populations of diploid and autotetraploid Heuchera grossulariifolia over three years. Our results demonstrate consistent directional selection for increasing scape length and floral display in both diploid and tetraploid populations. In contrast, selection acting on flowering phenology varied across year and ploidy. Specifically, selection was found to favor late-flowering diploids in 2001 and 2002 but early-flowering tetraploids in 2003. We investigated the mechanistic basis of divergent selection for flowering phenology in 2003 by estimating the relationship between plant flowering phenology and the probability of intercytotype pollinator movement. The results demonstrated that less divergent tetraploids were significantly more likely to experience intercytotype flights than were more divergent tetraploids. This result is consistent with the pattern of phenotypic selection observed. Taken together, our results suggest that divergence of polyploids and their diploid progenitors may be driven by a process analogous to reinforcement whereby selection favors phenotypes that reduce the probability of intercytotype matings with reduced fertility. PMID- 16261731 TI - Ecological factors influencing tetraploid speciation in snow buttercups (Ranunculus Adoneus): niche differentiation and tetraploid establishment. AB - Chromosome doubling plays an important role in generating new species of flowering plants. However, reproductive incompatibilities between newly formed tetraploid plants and their diploid progenitors are expected to create a significant barrier to the persistence and establishment of neopolyploid populations. Ecological differentiation can reduce this barrier via prezygotic isolation arising from spatial separation. Alternatively, superior viability or fecundity of neotetraploid plants might compensate for the reproductive cost of incompatible pollen from diploid neighbors. The performance of plants of both cytotypes can be assessed in their respective habitats through reciprocal transplants, although such experiments have not been used previously in the study of tetraploid speciation. We used a series of seed and seedling transplant experiments to assess ecological differentiation and competitive ability during early establishment phases for tetraploid and diploid forms of the snow buttercup (Ranunculus adoneus). At two sites, seeds from diploids and tetraploids had similar germination probabilities. Tetraploid snow buttercup seedlings had a significant growth advantage in a controlled environment chamber experiment. However, in the field diploid and tetraploid buttercup seedlings did not differ consistently in survival or growth, nor did the two cytotypes show reciprocal advantages in performance, as expected if ecological differentiation has occurred. At the seed and seedling stages, neither niche differentiation nor tetraploid competitive superiority appears sufficient to explain neotetraploid success in the presence of their diploid progenitors. PMID- 16261732 TI - Variation in asexual lineage age in Potamopyrgus antipodarum, a New Zealand snail. AB - Asexual lineages are thought to be subject to rapid extinction because they cannot generate recombinant offspring. Accordingly, extant asexual lineages are expected to be of recent derivation from sexual individuals. We examined this prediction by using mitochondrial DNA sequence data to estimate asexual lineage age in populations of a freshwater snail (Potamopyrgus antipodarum) native to New Zealand and characterized by varying frequency of sexual and asexual individuals. We found considerable variation in the amount of genetic divergence of asexual lineages from sexual relatives, pointing to a wide range of asexual lineage ages. Most asexual lineages had close genetic ties (approximately 0.1% sequence divergence) to haplotypes found in sexual representatives, indicating a recent origin from sexual progenitors. There were, however, two asexual clades that were quite genetically distinct (> 1.2% sequence divergence) from sexual lineages and may have diverged from sexual progenitors more than 500,000 years ago. These two clades were found in lakes that had a significantly lower frequency of sexual individuals than lakes without the old clades, suggesting that the conditions that favor sex might select against ancient asexuality. Our results also emphasize the need for large sample sizes and spatially representative sampling when hypotheses for the age of asexual lineages are tested to adequately deal with potential biases in age estimates. PMID- 16261733 TI - The genetic basis for fruit odor discrimination in Rhagoletis flies and its significance for sympatric host shifts. AB - Rhagoletis pomonella (Diptera: Tephritidae) use volatile compounds emitted from the surface of ripening fruit as important chemosensory cues for recognizing and distinguishing among alternative host plants. Host choice is of evolutionary significance in Rhagoletis because these flies mate on or near the fruit of their respective host plants. Differences in host choice based on fruit odor discrimination therefore result in differential mate choice and prezygotic reproductive isolation, facilitating sympatric speciation in the absence of geographic isolation. We test for a genetic basis for host fruit odor discrimination through an analysis of F2 and backcross hybrids constructed between apple-, hawthorn-, and flowering dogwood-infesting Rhagoletis flies. We recovered a significant proportion (30-65%) of parental apple, hawthorn, and dogwood fly response phenotypes in F2 hybrids, despite the general failure of F1 hybrids to reach odor source spheres. Segregation patterns in F2 and backcross hybrids suggest that only a modest number of allelic differences at a few loci may underlie host fruit odor discrimination. In addition, a strong bias was observed for F2 and backcross flies to orient to the natal fruit blend of their maternal grandmother, implying the existence of cytonuclear gene interactions. We explore the implications of our findings for the evolutionary dynamics of sympatric host race formation and speciation. PMID- 16261734 TI - Intralocus sexual conflict and the genetic architecture of sexually dimorphic traits in Prochyliza xanthostoma (Diptera: Piophilidae). AB - Because homologous traits of males and females are likely to have a common genetic basis, sex-specific selection (often resulting from sexual selection on one sex) may generate an evolutionary tug-of-war known as intralocus sexual conflict, which will constrain the adaptive divergence of the sexes. Theory suggests that intralocus sexual conflict can be mitigated through reduction of the intersexual genetic correlation (rMF), predicting negative covariation between rMF and sexual dimorphism. In addition, recent work showed that selection should favor reduced expression of alleles inherited from the opposite-sex parent (intersexual inheritance) in traits subject to intralocus sexual conflict. For traits under sexual selection in males, this should be manifested either in reduced maternal heritability or, when conflict is severe, in reduced heritability through the opposite-sex parent in offspring of both sexes. However, because we do not know how far these hypothesized evolutionary responses can actually proceed, the importance of intralocus sexual conflict as a long-term constraint on adaptive evolution remains unclear. In this study, we investigated the genetic architecture of sexual and nonsexual morphological traits in Prochyliza xanthostoma. The lowest rMF and greatest dimorphism were exhibited by two sexual traits (head length and antenna length) and, among all traits, the degree of sexual dimorphism was correlated negatively with rMF. Moreover, sexual traits exhibited reduced maternal heritabilities, and the most strongly dimorphic sexual trait (antenna length) was heritable only through the same-sex parent in offspring of both sexes. Our results support theory and suggest that intralocus sexual conflict can be resolved substantially by genomic adaptation. Further work is required to identify the proximate mechanisms underlying these patterns. PMID- 16261735 TI - Parallel evolution in ecological and reproductive traits to produce cryptic damselfly species across the holarctic. AB - The damselfly genus Enallagma originated in the Nearctic, and two Nearctic lineages recently underwent radiations partly associated with multiple independent habitat shifts from lakes dominated by fish predators into lakes dominated by dragonfly predators. A previous molecular study of four Palearctic morphospecies and all representative Nearctic species identified the presence of two cryptic species sets, with each set having Palearctic and Nearctic representatives. However, the cryptic species within each set are not sibling species. Here, we present quantitative data on ecologically important larval morphologies and behaviors involved in predator avoidance and on adult male morphological structures involved in mate recognition to quantify the phenotypic relationships among these cryptic species sets. For the adult stage, our data indicate strong parallel evolution of the structures involved in specific mate recognition-the male cerci. For the larval stage, morphometric analyses show that the Palearctic species evolved a nearly identical morphology to the sibling-clade members in the Nearctic that live in waters where dragonflies are the top predators. This implicates the importance of dragonfly predation in the history of the Palearctic clade. Behavioral analyses suggest population differentiation in response to the actual predator environment in the Palearctic clade, consistent with the species differentiation seen in the Nearctic. Our results suggest parallel evolution of adult traits that influence specific mate choice and larval traits that influence ecological performance underlie the striking similarity of Enallagma species across continents. This concurrent parallel evolution in both stages of a complex life cycle, especially when both stages do not share the same selective environment, may be a very unusual mechanism generating cryptic species. PMID- 16261736 TI - Have male and female genitalia coevolved? A phylogenetic analysis of genitalic morphology and sexual size dimorphism in web-building spiders (Araneae: Araneoidea). AB - Sexual size dimorphism (SSD) can strongly influence the evolution of reproductive strategies and life history. If SSD is extreme, and other characters (e.g., genitalic size) also increase with size, then functional conflicts may arise between the sexes. Spiders offer an excellent opportunity to investigate this issue because of their wide range of SSD. By using modern phylogenetic methods with 16 species of orb-weaving spiders, we provide strong evidence for the "positive genitalic divergence" model, implying that sexual genitalic dimorphism (SGD) increases as SSD increases. This pattern is supported by an evolutionary mismatch between the absolute sizes of male and female genitalia across species. Indeed, our findings reveal a dramatic reversal from male genitalia that are up to 87x larger than female genitalia in size-monomorphic species to female genitalia that are up to 2.8x larger in extremely size-dimorphic species. We infer that divergence in SGD could limit SSD both in spiders, and potentially in other taxa as well. Further, male and female body size, as well as male and female genitalia size, are decoupled evolutionarily. Finally, we show a negative scaling (hypoallometry) of male and female genitalic morphology within sexes. Evolutionary forces specific to each sex, such as larger female size (increased fecundity) or smaller male size (enhanced mate-searching ability), may be balanced by stabilizing selection on relative genitalic size. PMID- 16261738 TI - Low survival of parasite chicks may result from their imperfect adaptation to hosts rather than expression of defenses against parasitism. AB - Host parents exhibit a variety of behaviors toward avian brood parasites, but not all of their actions have necessarily evolved in response to costs imposed by parasites. To investigate whether common waxbills (Estrilda astrild) have evolved defenses specifically against parasitic pin-tailed whydahs (Vidua macroura), I studied the specificity and flexibility of host behaviors toward nestlings at two sites that differed significantly in parasitism rates and intensities. I focused on documenting nestling survival because V. macroura young match the elaborate gape morphology of E. astrild nestlings, a pattern that suggests hosts may possess unique defenses against parasite chicks. Parasite young survived significantly worse than host young in mixed broods. However, this apparent discrimination was not associated with parasitism risk as would be expected if defenses had evolved specifically to counter parasitism. Parasite young may have survived poorly compared to host young because individual chicks were less able to stimulate sufficient care from foster parents or because they were more susceptible to nestling competition, disease, or reduced provisioning by hosts. Mortality may have also been exacerbated by poor timing of parasite egg laying. In nonparasitized and parasitized nests, rates of nestling survival were similar, further suggesting that parenting behaviors that result in chick mortality did not evolve solely in response to parasite young. In addition, orange-breasted waxbills (Amandava subflava) and zebra finches (Taeniopygia guttata), rarely parasitized and nonparasitized relatives of E. astrild, experience similar levels of nestling mortality presumably as a result of phylogenetically widespread parenting strategies. Despite the similarity of parasitic V. macroura nestlings and E. astrild nestlings, I found no evidence that E. astrild parents possess defenses that allow for specific discrimination against parasite chicks during the nestling period. Rather than being subject to host defenses evolved in an arms race, Vidua chicks may simply be imperfectly adapted to life in the nests of their hosts. PMID- 16261737 TI - Phylogenetic analysis of ecomorphological divergence, community structure, and diversification rates in dusky salamanders (Plethodontidae: Desmognathus). AB - An important dimension of adaptive radiation is the degree to which diversification rates fluctuate or remain constant through time. Focusing on plethodontid salamanders of the genus Desmognathus, we present a novel synthetic analysis of phylogeographic history, rates of ecomorphological evolution and species accumulation, and community assembly in an adaptive radiation. Dusky salamanders are highly variable in life history, body size, and ecology, with many endemic lineages in the southern Appalachian Highlands of eastern North America. Our results show that life-history evolution had important consequences for the buildup of plethodontid-salamander species richness and phenotypic disparity in eastern North America, a global hot spot of salamander biodiversity. The origin of Desmognathus species with aquatic larvae was followed by a high rate of lineage accumulation, which then gradually decreased toward the present time. The peak period of lineage accumulation in the group coincides with evolutionary partitioning of lineages with aquatic larvae into seepage, stream edge, and stream microhabitats. Phylogenetic simulations demonstrate a strong correlation between morphology and microhabitat ecology independent of phylogenetic effects and suggest that ecomorphological changes are concentrated early in the radiation of Desmognathus. Deep phylogeographic fragmentation within many codistributed ecomorph clades suggests long-term persistence of ecomorphological features and stability of endemic lineages and communities through multiple climatic cycles. Phylogenetic analyses of community structure show that ecomorphological divergence promotes the coexistence of lineages and that repeated, independent evolution of microhabitat-associated ecomorphs has a limited role in the evolutionary assembly of Desmognathus communities. Comparing and contrasting our results to other adaptive radiations having different biogeographic histories, our results suggest that rates of diversification during adaptive radiation are intimately linked to the degree to which community structure persists over evolutionary time. PMID- 16261739 TI - Can selection by an ectoparasite drive a population of red crossbills from its adaptive peak? AB - The bill structures of different call types of red crossbills (Loxia curvirostra complex) in western North America usually approximate the predicted optima for foraging on single species of conifers. One clear exception is the call type in the South Hills, Idaho, that is coevolving in an evolutionary arms race with Rocky Mountain lodgepole pine (Pinus contorta ssp. latifolia). Although South Hills crossbills forage only on the cones of these lodgepole pines, their average bill depth is smaller than that predicted to be optimal. Because preliminary data showed that large-billed males were more likely to exhibit symptoms of ectoparasitic mite (Knemidokoptes jamaicensis) infestation, the goal of our study was to further quantify the incidence of mite infestation and determine whether selection by mites may have favored smaller-billed crossbills and thus driven crossbills away from the foraging optimum. We estimated annual survival of both infected and uninfected South Hills crossbills using program MARK, which allows for auxiliary variables such as bill size and sex to be included in survival analyses. Mite infestation depressed crossbill survival and, especially for males, caused directional selection against larger-billed individuals. Such selection may explain why South Hills crossbills have smaller bills than the optimum and why average bill size for males has decreased from 1998 to 2003. This selection may also explain why the degree of sexual size dimorphism has decreased by nearly 50% since 1998. PMID- 16261740 TI - Speciational history of Australian grass finches (Poephila) inferred from thirty gene trees. AB - Multilocus genealogical approaches are still uncommon in phylogeography and historical demography, fields which have been dominated by microsatellite markers and mitochondrial DNA, particularly for vertebrates. Using 30 newly developed anonymous nuclear loci, we estimated population divergence times and ancestral population sizes of three closely related species of Australian grass finches (Poephila) distributed across two barriers in northern Australia. We verified that substitution rates were generally constant both among lineages and among loci, and that intralocus recombination was uncommon in our dataset, thereby satisfying two assumptions of our multilocus analysis. The reconstructed gene trees exhibited all three possible tree topologies and displayed considerable variation in coalescent times, yet this information provided the raw data for maximum likelihood and Bayesian estimation of population divergence times and ancestral population sizes. Estimates of these parameters were in close agreement with each other regardless of statistical approach and our Bayesian estimates were robust to prior assumptions. Our results suggest that black-throated finches (Poephila cincta) diverged from long-tailed finches (P. acuticauda and P. hecki) across the Carpentarian Barrier in northeastern Australia around 0.6 million years ago (mya), and that P. acuticauda diverged from P. hecki across the Kimberley Plateau-Arnhem Land Barrier in northwestern Australia approximately 0.3 mya. Bayesian 95% credibility intervals around these estimates strongly support Pleistocene timing for both speciation events, despite the fact that many gene divergences across the Carpentarian region clearly predated the Pleistocene. Estimates of ancestral effective population sizes for the basal ancestor and long tailed finch ancestor were large (about 521,000 and about 384,000, respectively). Although the errors around the population size parameter estimates are considerable, they are the first for birds taking into account multiple sources of variance. PMID- 16261742 TI - Translating research into practice: intervening to promote better care. AB - Promoting better healthcare means tackling a slew of challenges such as obesity, smoking, and substance abuse. Researchers at the annual "Translating Research Into Practice" conference revealed how they are finding new ways to successfully address these challenges-breaking down barriers and providing solutions. PMID- 16261743 TI - VHA, Inc., introduces new data-collection tool allowing national and regional comparisons in real-time. AB - VHA, Inc., has rebuilt its online comparative clinical measurement (CCM) tool to help get hospital members on the same page in terms of clinical measurement in real-time. In June, it introduced a new version (CCM 2005) that includes the six new clinical goals or "changes of care" outlined late last year by the Institute for Healthcare Improvement in its 100,000 Lives campaign. PMID- 16261741 TI - Association of ploidy and sexual system in Lycium californicum (Solanaceae). AB - In North American Lycium (Solanaceae), the evolution of gender dimorphism has been proposed as a means of restoring outcrossing after polyploidization causes the loss of self-incompatibility. Previous studies of this process in Lycium focused on comparisons between species that differ in ploidy. We examined intraspecific variation in floral morphology and DNA content in populations of L. californicum to determine correlations between sexual system and cytotype. We also used nuclear ITS and GBSSI sequence data to determine whether diploid and polyploid forms represent the same phylogenetic species, and the phylogeographic relationships among populations and ploidy levels. Within populations, no variation in ploidy was found, although among populations there was a perfect correspondence between sexual system and cytotype. Diploid populations were all hermaphroditic, whereas tetraploid populations were all gender dimorphic. There was no clear geographic pattern to the occurrence of diploid and tetraploid forms. Phylogenetic analysis confirms that L. californicum, regardless of ploidy, forms a monophyletic group within the genus Lycium. Sequences from diploid and polyploid individuals did not form reciprocally monophyletic clades, indicating either multiple gains of polyploidy, ongoing gene flow between cytotypes, or lack of lineage sorting since the evolution of polyploidy. The correspondence between ploidy and sex expression is consistent with the hypothesis that polyploidization triggers the evolution of gender dimorphism in this and other Lycium species. PMID- 16261744 TI - Engineering a high-quality healthcare system. PMID- 16261745 TI - New Pennsylvania report provides insights on hospital-acquired infections. PMID- 16261746 TI - AHA recognizes hospitals for community health services. PMID- 16261747 TI - It is time for otolaryngologists to perform EGD. PMID- 16261748 TI - Glomus tympanicum. PMID- 16261749 TI - Nasopharyngeal teratoma in an adult. PMID- 16261751 TI - Schwannoma of the larynx. PMID- 16261750 TI - Vocal fold polyp in a professional singer. PMID- 16261752 TI - Reconstruction of the eyebrow. PMID- 16261753 TI - Prosthetic rehabilitation of large sinonasal-orbital and palatal defects. PMID- 16261754 TI - Neuroendocrine adenoma of the middle ear. PMID- 16261755 TI - Improving billing practices through peer-group feedback. PMID- 16261756 TI - Tonsillectomy without headlights: a unique solution. PMID- 16261757 TI - Nonsurgical home treatment of middle ear effusion and associated hearing loss in children. Part I: clinical trial. AB - We conducted a randomized, controlled clinical trial to investigate the efficacy of treatment of persistent middle ear effusion (MEE) and associated hearing loss with a modified Politzer device used in the home setting over a 7-week period. Efficacy was determined by comparing pre-and posttherapy air-conduction thresholds, tympanometric peak pressures, and otoscopic findings. The study group was made up of 94 children (174 ears), aged 4 to 11 years, who had at least a 2 month history of MEE and associated hearing loss. At study's end, patients in the treatment group experienced statistically significant improvements in all measured outcomes; no significant improvements were seen in the control group in all measured outcomes. At study's end, the hearing sensitivity of 73.9% of the treated ears was within normal limits, compared with only 26.7% of the control ears. These findings demonstrate that home treatment of children with persistent MEE and associated hearing loss with the modified Politzer device is highly efficacious. PMID- 16261758 TI - Vertigo and motion sickness. Part I: vestibular anatomy and physiology. AB - Control of the symptoms of vertigo and motion sickness requires consideration of the neurophysiology of areas both intrinsic and extrinsic to the vestibular system proper. We review the essential anatomy and physiology of the vestibular system and the associated vomiting reflex. PMID- 16261759 TI - Endoscopic resection of a venous hemangioma of the optic nerve sheath. AB - We describe a case of a venous hemangioma that arose in the optic nerve sheath of the orbital apex in a 28-year-old man who presented with progressive vision loss. To the best of our knowledge, this is the first reported case of a venous hemangioma occurring at this location. A definitive diagnosis and partial excision was achieved via an endoscopic transsphenoethmoid approach with interactive, computer-assisted, frameless stereotactic surgical navigation. The intervention resulted in minimal morbidity, demonstrating yet again that this surgical approach is a safe and effective way to treat lesions of the orbital apex. PMID- 16261760 TI - Primary chordoma of the lateral nasal wall: case report and review. AB - Chordomas are malignant, nonepithelial neoplasms derived from notochordal tissue. A primary chordoma of the nasal cavities and paranasal sinuses is extremely rare compared with clival chordomas, which often present as nasal masses after spreading anteriorly. Only a few cases of primary chordoma of the nasal cavities and paranasal sinuses have been reported in the literature. We report a case of a primary chordoma of the lateral nasal wall. Pathologic diagnosis was obtained using an intranasal endoscopic excision. As in our patient, a primary chordoma of the nasal cavity or paranasal sinuses may present with symptoms related to mechanical obstruction secondary to the tumor mass. We summarize our case in the context of the other reported cases, and we discuss treatment options, natural history, and prognosis. PMID- 16261761 TI - Acinic cell carcinoma of the parotid gland: a 15-year review limited to a single surgeon at a single institution. AB - The course of acinic cell carcinoma of the parotid gland following surgical and nonsurgical interventions is variable. The objective of this study was to report our experience in treating this disease and to evaluate the factors that might be involved in the treatment of the tumor and the prognosis of the patient. To limit the contributory variables that are usually found in most studies, we included only those patients (n = 11) who had been treated by a single surgeon at a single institution from 1988 through 2003. Hospital and office records were evaluated for demographic information, signs and symptoms, treatment modalities, pathology, and outcomes. Additional follow-up data were obtained through telephone interviews. For the most part, treatment included either superficial parotidectomy or total parotidectomy with facial nerve preservation; 1 patient with coexisting adenocarcinoma underwent a more radical procedure, and 4 patients underwent adjuvant radiation therapy. The most prevalent morphologic pattern of these tumors was microcystic. Follow-up ranged from 1 year and 3 months to 10 years and 9 months (mean: 4 yr and 11 mo). During that time, we found no recurrences of acinic cell carcinoma and no evidence of metastatic disease. Therefore, we conclude that acinic cell carcinoma can be successfully treated with a superficial or total parotidectomy with sparing of the facial nerve. Radiation therapy may provide adjunctive benefit. PMID- 16261762 TI - Implant exposure techniques at second stage surgery. Interview. PMID- 16261763 TI - Developmental competence and gene expression in preimplantation bovine embryos derived from somatic cell nuclear transfer using different donor cells. AB - This study compared the developmental competence of somatic cell nuclear transfer (SCNT) embryos reconstructed with different donor cells and analysed gene expression in the resulting embryos. Bovine fetal/adult ear fibroblasts and cumulus cells were used as donor cells and the developmental competence of the reconstructed embryos was monitored. The cell number and allocation in blastocysts were determined by differential staining. The Bax, E-cad, IF-tau, Hsp (heat shock protein) 70, Igf2r (insulin-like growth factor 2 receptor), DNMT (DNA methyltransferase) 1 and Mash (mammalian achaete-scute homologue) 2 genes were selected for gene expression analysis. The relative abundance (ratio to GAPDH mRNA) of gene transcripts in blastocysts was measured by semiquantitative reverse transcription-polymerase chain reaction. In experiment 1, development of SCNT preimplantation embryos and the cell numbers of inner cell masses and trophoblasts were not different among SCNT embryos derived from different cell types. In experiment 2, the relative expression of GAPDH and Hsp 70 transcripts was similar in all embryos. The expression of Bax, Igf2r and Mash2 transcripts was significantly increased in SCNT embryos reconstructed with adult fibroblasts. The E-cad transcript levels were reduced in SCNT embryos reconstructed with fetal fibroblasts. Relative abundance of DNMT1 in SCNT embryos derived from fetal fibroblasts was increased, and IF-tau expression in SCNT embryos derived from cumulus cells was increased. In conclusion, depending on the type of donor cells, preimplantation SCNT embryos displayed marked differences in gene expression. This may affect the developmental competence of SCNT embryos reconstructed with different cell types after implantation or during fetal growth in vivo. PMID- 16261764 TI - Molecular and ultrastructural studies of the sperm chromatin from Triturus cristatus. AB - The study of nuclear molecular architecture during gametogenesis represents one approach towards the deciphering of the molecular organization of eukaryotic chromatin. During spermatogenesis, chromatin undergoes several dynamic transitions, which are often associated with important changes not only in its physical conformation but even in its composition and structure. Dynamic alterations in chromatin structure mediated by postsynthetic histone modification and DNA methylation constitute a major regulatory mechanism of gene function of eukaryotes. Using transmission electron microscopy and molecular investigations, some peculiar aspects of chromatin organization and evolution in spermatogenesis of the crested newt Triturus cristatus were investigated. We focused our investigations on the dynamics of chromatin structure after treatment with TSA (a histone deacetylase inhibitor). PMID- 16261765 TI - Hyaluronan and its binding proteins in the epithelium and intraluminal fluid of the bovine oviduct. AB - Hyaluronan (HA) is involved in several important steps of sperm storage and of fertilization. This study investigates the presence and concentration of HA in oviductal fluid (ODF), together with the localization of HA and the presence of hyaluronan-binding proteins (HABPs) in the oviductal epithelium of normally cycling dairy heifers and cows. The concentration and amount of HA in ODF, collected over the course of several oestrous cycles via catheters placed in the isthmic and ampullar tubal segments, were measured using an ELISA. The concentration and amount of HA in ODF did not vary significantly between these anatomical regions, nor between the stages of the oestrous cycle (p > 0.05), although the amount of HA seemed to peak during oestrous. The most HA per day (2.9 +/- 0.64 microg, least square mean +/- SEM) was produced on the day of ovulation, whereas the lowest amount (1.25 +/- 0.68 microg) was produced 4 days before ovulation. To investigate the localization of HA, tissue samples were retrieved at well-defined stages of the oestrous cycle and from corresponding regions of the oviduct. Sections and protein extracts from the tissue samples were studied histochemically using biotinylated HABP and immunoblotted with fluorescein isothiocyanate (FITC)-HA, respectively. Presence of HA labelling in the oviductal epithelium was restricted to the sperm reservoir, a localization that seemed to be cycle-independent. The immunoblotting of samples from the lining epithelium revealed seven bands of HABPs. We confirm that the bovine oviduct produces HA and its binding proteins, and that HA is mainly localized to the epithelium of the sperm reservoir. PMID- 16261766 TI - Behavior of delta-tubulin during spindle formation in Xenopus oocytes: requirement of cytoplasmic dynein-dependent translocation. AB - Vertebrate oocytes do not contain centrosomes and therefore form an acentrosomal spindle during oocyte maturation. gamma-Tubulin is known to be essential for nucleation of microtubules at centrosomes, but little is known about the behaviour and role of gamma-tubulin during spindle formation in oocytes. We first observed sequential localization of gamma-tubulin during spindle formation in Xenopus oocytes. gamma-Tubulin assembled in the basal regions of the germinal vesicle (GV) at the onset of germinal vesicle breakdown (GVBD) and remained on the microtubule-organizing centre (MTOC) until a complex of the MTOC and transient-microtubule array (TMA) reached the oocyte surface. Prior to bipolar spindle formation, oocytes formed an aggregation of microtubules and gamma tubulin was concentrated at the centre of the aggregation. At the late stage of bipolar spindle formation, gamma-tubulin accumulated at each pole. Anti-dynein antibody disrupted the localization of gamma-tubulin, indicating that the translocation described above is dependent on dynein activity. We finally revealed that XMAP215, a microtubule-associated protein cooperating with gamma tubulin for the assembly of microtubules, but not gamma-tubulin, was phosphorylated during oocyte maturation. These results suggest that gamma-tubulin is translocated by dynein to regulate microtubule organization leading to spindle formation and that modification of the molecules that cooperate with gamma tubulin, but not gamma-tubulin itself, is important for microtubule reorganization. PMID- 16261767 TI - Effects of canine serum collected from dogs at different estrous cycle stages on in vitro nuclear maturation of canine oocytes. AB - Canine oocytes are ovulated at prophase of the first meiotic division and undergo maturation in the distal part of the oviduct for at least 48-72 h. Because of these differences from other domestic mammals, the efficiency of in vitro maturation (IVM) of canine oocyte is very low. The present study was conducted to evaluate the effects of canine serum on IVM of canine oocytes recovered from ovaries in various reproductive states (follicular, luteal or anestrous stages). Oocytes were recovered by mincing ovaries from bitches presented for ovariohysterectomy at various stages of the estrous cycle. Heat-inactivated canine serum was prepared with blood taken from dogs at the anestrous, estrous or diestrous stage of the estrous cycle as determined by progesterone concentration and vaginal cytology. Oocytes were cultured for 72 h in tissue culture medium (TCM)-199 supplemented with 10% canine anestrous, estrous or diestrous serum or fetal bovine serum (FBS) (experiment 1), or supplemented with 0 (control), 5%, 10% or 20% canine estrous serum (experiment 2). In experiment 1, IVM of oocytes collected at the follicular stage of the estrous cycle to metaphase II (MII) stage was higher (p < 0.05) with canine estrous serum (14.2%) than with canine anestrous (5.2%) or diestrous serum (6.3%), FBS (2.2%) or in the control (2.2%). In experiment 2, oocytes collected at the follicular stage of the estrous cycle cultured in TCM-199 with 10% canine estrous serum showed a higher maturation rate to MII stage (13.5%, p < 0.05) compared with those cultured with 5% (1.3% MII) or 20% canine estrous serum (5.1% MII) or the control (2.7% MII). In conclusion, our results demonstrate that supplementing culture medium with 10% canine estrous serum improves IVM of canine follicular stage oocytes. PMID- 16261769 TI - Ultrastructural study of spermatozoa of the paddlefish, Polyodon spathula. AB - Paddlefish, Polyodon spathula, spermatozoa were examined by transmission electron microscopy. Their structure has the same characteristic architectural features as sturgeon spermatozoa. Paddlefish spermatozoa are of the primitive type and consist of a rod-shaped head, a midpiece and a long flagellum. The head is about 5.15 mm in length and contains the nucleus and an apical acrosomal complex. Inside the nucleus there are three nuclear channels that begin in the subacrosomal area and have a triple helical arrangement. An nuclear fossa is present centrally, at the posterior end of the nucleus. The midpiece contains a pair of centrioles in a perpendicular arrangement, mitochondria and a narrow cytoplasmic sleeve. The flagellum has a central axoneme with a 9 + 2 pattern and two lateral projections or fins. PMID- 16261768 TI - In vitro culture and mtDNA fate of ibex-rabbit nuclear transfer embryos. AB - Rabbit oocyte can be used as the recipient in interspecies somatic cell nuclear transfer (iSCNT). This work was undertaken in order to study the developmental competence of Capra ibex somatic cells reprogrammed by rabbit oocytes and the fate of mitochondria in iSCNT embryos. Metaphase II (MII) oocytes from superovulated rabbit were used as nuclear recipients. The nuclear donors were Capra ibex somatic cells with different proliferative status: population doubling time (PDL) = 15 +/- 2 (group 1), 35 +/- 2 (group 2), 55 +/- 2 (group 3) and 70 +/ 2 (group 4). Oocytes reconstructed with electrical pulses (2.1kV/cm, 10 micros, 2 times) were activated (1.4kV, 20 micros, 2 times) and then cultured in Medium199 containing 10% fetal bovine serum at 38.5 degrees C, 5% CO2 in air. In groups 1, 2, 3 and 4, the fusion rates were 35.83%, 66.03%, 65.40% and 35.35%, respectively. Similar cleavage rates were observed among the four groups. However, the developmental potential to morula/blastocyst from early nuclear donor embryos (16.42%/10.45%) was significantly higher (p < 0.05) than in terminal donor embryos (9.52%/3.81%). Polymerase chain reaction analysis of the mitochondrial (mt) DNA cytb gene demonstrated that mtDNAs from ibex and rabbit could be detected at various developmental stages before implantation. In conclusion, our results provide some original information about rescuing Capra ibex using the iSCNT technique. These results indicate that: (1) enucleated rabbit oocytes make Capra ibex fibroblast nuclei reprogramme; (2) the proliferative status of donor cells affects the efficiency of iSCNT; and (3) rabbit ooplasm rescues the donor-derived mtDNAs, resulting in mtDNA heteroplasmy before implantation. PMID- 16261770 TI - Effects of oocyte collection time and injection position on pronucleus formation and blastocyst development in round spermatid injection in mouse. AB - The injection of spermatozoa into mouse, human and rabbit oocytes at specific times and positions can result in different rates of viable embryo development. However, it is not clear how the timing and position of round spermatid injection (ROSI) affect pronucleus (PN) formation and blastocyst development of mice. First, we determined the changes in relative position of the first polar body and the spindle, carried out ROSI from 11.5 to 13 h post-hCG administration, then activated by Sr2+, and finally compared the development of ROSI zygotes, including the formation of pronuclei and development of blastocyst. Between 11.5 and 13 h post-hCG administration, the rate of 2PN formation by ROSI at 3 o'clock was the highest among all treated oocytes. Moreover, the blastocyst rate of zygotes with two pronuclei (2PN) was up to 27.41%. These results suggest that the time and position of ROSI can significantly influence the formation of 2PN, that the rates of 2PN formation are closely correlated with blastocyst formation and that the formation of 2PN is necessary for later embryo development. PMID- 16261771 TI - Septate junctions mediate the barrier to paracellular permeability in sea urchin embryos. AB - In sea urchin embryos, blastula formation occurs between the seventh and tenth cleavage and is associated with changes in the permeability properties of the epithelium although the structures responsible for mediating these changes are not known. Tight junctions regulate the barrier to paracellular permeability in chordate epithelia; however, the sea urchin blastula epithelium lacks tight junctions and instead possesses septate junctions. Septate junctions are unique to non-chordate invertebrate cell layers and have a characteristic ladder-like appearance whereby adjacent cells are connected by septa. To determine the function of septate junctions in sea urchin embryos, the permeability characteristics of the embryonic sea urchin epithelia were assessed. First, the developmental stage at which a barrier to paracellular permeability arises was examined and found to be in place after the eighth cleavage division. The mature blastula epithelium is impermeable to macromolecules; however, brief depletion of divalent cations renders the epithelium permeable. The ability of the blastula epithelium to recover from depletion of divalent cations and re-establish a barrier to paracellular permeability using fluorescently labelled lectins was also examined. Finally, septate junction structure was examined in embryos in which the permeability status of the epithelium was known. The results provide evidence that septate junctions mediate the barrier to paracellular permeability in sea urchin embryos. PMID- 16261772 TI - Role of prolactin in nuclear maturation and ovulation in amphibian oocytes. AB - The present study was undertaken to determine the effect of prolactin (Prl) on Bufo arenarum oocyte maturation and ovulation, two characteristic events of the breeding period, the stage of the sexual cycle in which gamete growth is complete. We observed that Prl, at the doses assayed, did not affect nuclear maturation per se. In addition, when follicles were pretreated with Prl and progesterone was later added to the medium as a physiological nuclear maturation inducer, the percentage of germinal vesicle breakdown obtained with the steroid was unaffected by Prl. The analysis of the in vitro ovulation process demonstrated that pituitary homologous homogenate (PHH) produced a dose- and month-dependent stimulating effect. The maximum percentage of ovulated oocytes was obtained from the end of July to October, the period in which oviposition naturally occurs. Prl by itself did not affect the ovulation process, but when both the hormone and PHH were present in the incubation medium, a significant increase in ovulated oocytes was observed. The results suggest that Prl does not participate in oocyte maturation; however, its presence in the incubation medium would increase oocyte sensitivity to the action of the physiological ovulation inducers. PMID- 16261773 TI - Effect of exogenous carbohydrates in a serum-free culture medium on the development of in vitro matured and fertilized porcine embryos. AB - This study was conducted to examine the effect of energy substrates in a serum free culture medium on in vitro development of porcine embryos. Presumptive zygotes derived from in vitro fertilization were cultured in glucose-free North Carolina State University (NCSU)-23 medium with glucose, pyruvate, fructose and lactate added to the culture medium singly or in various combinations. In experiment 1, a higher percentage of embryos cleaved (53-63% vs 10-13%) and developed to the blastocyst stage (18-27% vs 0) after the single addition of glucose (5.6 mM), pyruvate (0.5 mM) or lactate (10 mM) than with no energy substrate addition or the addition only of fructose (5.6 mM). In experiment 2, the addition of pyruvate and lactate resulted in higher blastocyst formation (25%) than other combinations (6-22%), while the addition of glucose and pyruvate significantly inhibited blastocyst formation. Increasing lactate concentration, as a single energy supplement, from 5 to 20 mM significantly improved blastocyst formation (7% vs 14-18%), while no benefit was achieved from increasing pyruvate concentration up to 2 mM (experiment 3). Glucose-free NCSU-23 medium supplemented with 0.5 mM pyruvate and 5 mM lactate significantly improved blastocyst formation (28% vs 17%) compared with NCSU-23 medium supplemented with 5.6 mM glucose (experiment 4). In conclusion, pyruvate and lactate are preferable energy substrates to support in vitro development of porcine embryos cultured in a serum free NCSU-23 medium. PMID- 16261774 TI - Comparison of the effects of desflurane and sevoflurane on the QTc interval and QT dispersion. AB - OBJECTIVE: The effects of desflurane and sevoflurane on QT interval and QT dispersion have been investigated in a prospective, double-blind, randomized study of patients undergoing noncardiac surgery. INTERVENTIONS: Sixty American Society of Anaesthesiologists physical status I-II adult patients were randomly assigned to two groups. Anaesthesia was induced with inhalation of desflurane (desflurane group) or sevoflurane (sevoflurane group) in increasing concentrations to 3 minimal alveolar concentration level. The maintenance of anaesthesia was provided with 2 minimal alveolar concentration agents in both groups until the end of the study. Electrocardiogram, heart rate and blood pressure were recorded as follows: before premedication, before induction, 1 and 3 min after the induction of anaesthesia, after the administration of vecuronium and after the tracheal intubation. The induction times and the complications were recorded. MEASUREMENTS AND RESULTS: The QTc interval was significantly more prolonged with desflurane than with sevoflurane at the first and third minute after the induction, and at the third minute after the administration of vecuronium. There were no significant differences in the QT dispersion between the two groups. Heart rate and blood pressure were found to be significantly higher in the desflurane group. CONCLUSION: The QTc interval was more prolonged with desflurane than sevoflurane, and QT dispersion was normal with both agents. PMID- 16261776 TI - Frequency of diastolic dysfunction in patients with sickle cell anaemia: a tissue Doppler imaging study. AB - OBJECTIVE: Although cardiac dilatation and systolic dysfunction have been well documented as a late manifestation, there are conflicting reports regarding the diastolic dysfunction, by using traditional echocardiographic methods in patients with sickle cell anaemia (SCA). The aim of this study was to investigate left ventricular diastolic function in SCA patients without congestive heart failure by using pulsed wave tissue Doppler imaging (TDI). METHODS: Thirty-one patients (18 men and 13 women, mean age: 26.4 +/- 8.5 years) with homozygous SCA without congestive heart failure and 31 healthy control subjects (19 men and 12 women, mean age: 26.5 +/- 7.6 years) were enrolled in the study. All patients and the controls underwent echocardiographic examination. Left ventricular diastolic function was evaluated by using both traditional pulsed wave Doppler (PWD) and TDI. RESULTS: The cardiac chamber dimensions, wall thicknesses, E and A transmitral peak diastolic velocities, systolic (Sm), early (Em) and late (Am) diastolic myocardial velocities were significantly increased in the patients compared to the controls. Diastolic dysfunction was found in 7 (22.6%) and 3 (9.7%) patients by PWD and TDI, respectively. Of the 7 patients with diastolic abnormalities, 4 patients with a restrictive pattern by PWD were found to be normal by TDI. CONCLUSION: Our results show that left ventricular diastolic dysfunction is not a frequent finding in patients with SCA without congestive heart failure and TDI is a more reliable and easy method to evaluate diastolic function. Patients with a restrictive pattern found by PWD should be reevaluated by using TDI. PMID- 16261775 TI - Apoptotic myocardial cell death in the setting of arrhythmogenic right ventricular cardiomyopathy. AB - BACKGROUND: Arrhythmogenic right ventricular cardiomyopathy (ARVC) is a primary heart muscle disease characterized by progressive atrophy of the right ventricular myocardium accompanied by fibro-fatty replacement. We examined whether the loss of myocardial cells in the setting of ARVC could result from cell death by apoptosis as demonstrated by the detection of DNA fragmentation and the expression of apoptosis regulatory proteins (e.g. CPP-32, Bcl-2, and Bax). METHODS: Specimens obtained from the right ventricular myocardium of 11 patients with ARVC (ARVC group) and 10 age-matched normal individuals (control group) were analysed. To identify individual cells undergoing apoptosis, paraffin sections were examined with the TdT-mediated dUTP-biotin nick end labelling method (TUNEL) and the rabbit polyclonal anti-single stranded DNA method (ssDNA). The apoptotic index was calculated as the percentage of nuclei staining positive by the TUNEL or ssDNA method. We also used immunohistochemical techniques to examine the levels of CPP-32, Bcl-2, and Bax expression. RESULTS: Apoptosis was detected in the ARVC group with a mean apoptotic index of 5.7 +/- 4.5% (ssDNA) and 23.8 +/- 7.5% (TUNEL), but not in the control group (p < 0.01). CPP-32 expression and Bax overexpression were observed in the ARVC group. However, Bcl-2 expression was not seen in either group. CONCLUSIONS: Apoptotic myocardial cell death occurs in the setting of ARVC and may contribute to the loss of myocardial cells. PMID- 16261777 TI - Time domain heart rate variability analysis in patients with thalassaemia major. AB - OBJECTIVE: Cardiac complication is one of the major causes of death in patients with thalassaemia major. Heart rate variability is a non-invasive index of neuronal modulation of heart rate. In this study, autonomic functions of the heart in a preclinical stage of heart disease were evaluated in 48 thalassaemic patients with time domain heart rate variability analysis; the control group consisted of 45 healthy subjects. METHODS AND RESULTS: Mean RR time in the study and control group was 0.73 +/- 0.1, and 0.82 +/- 0.1 ms respectively (p < 0.001). In the study group; SDNN, SDNN-i, SDANN-i, RMSSD, and PNN50 were found to be 167.4 +/- 86.2 ms, 153.9 +/- 108.1 ms, 111.4 +/- 60.4 ms, 108.9 +/- 86.7 ms, and 14.5 +/- 13.4%, respectively. Time domain parameters were significantly lower in the study group than the control group (p < 0.00 1). There was a significant positive correlation between the mean RR time and SDNN, SDNN-i, SDANN-i, RMSSD, and PNN50 (for the RMSSD p < 0.05, r = 0.31; for the others p = 0.000, and r values were 0.65, 0.65, 0.38, 0.37 for the SDNN, SDNN-i, SDANN-i, and PNN50, respectively). CONCLUSION: The analysis of heart rate variability might be helpful to detect cardiac complications in the preclinical stage of the cardiac involvement. PMID- 16261778 TI - Femoral access management: comparison between two different vascular closure devices after percutaneous coronary intervention. AB - OBJECTIVES: Several devices have been proposed as an alternative to manual compression (MC) for femoral access management (FAME) following catheterization. Although these devices allow earlier ambulation, they have not always been shown to reduce vascular complications. As a consequence, their cost efficacy is not obvious. METHODS: During MC a special catheter deployed temporarily within the artery to achieve haemostasis (Bio-DISC) (BD) was compared with an anchor collagen based system Angio-Seal (AS) among 463 consecutive patients undergoing PCI. We examined vascular or systemic complications, nursing time spent to puncture site management and patient's satisfaction. RESULTS: Relative contra indications to the use of vascular closure devices were encountered in 158 patients. There were no significant differences in baseline characteristics between the patients assigned to each of the 3 treatment groups. The deployment success rate was 98% for AS and 90% for BD (p = 0.037). Vascular complications occurred in 10.8%, 4.0% and 5.8% (p: NS) of MC, AS and BD patients, respectively. The longer sheath dwell time contributed to most of the complications in MC and BD. Nursing time spent for access management was 48.9 min in MC; 28.1 min in BD and 9.9 min in AS (p < 0.0001 ). Satisfaction score above 70 was noted in 46%, 86% and 92% of patients managed by MC, BD and AS, respectively. CONCLUSION: AS use is associated with fewer complications, improved patient well being and saves 39 minutes of nursing time. The additional cost of AS is justified when used in selected patients undergoing PCI. PMID- 16261779 TI - Serum uric acid levels as a predictor of in-hospital death in patients hospitalized for decompensated heart failure. AB - OBJECTIVES: The aim of this study was to determine the value of serum uric acid levels in predicting in-hospital mortality of chronic heart failure patients hospitalized for decompensation in spite of appropriate medical therapy. METHODS AND RESULTS: This study was conducted in patients who were admitted to our clinic between January 2003 and April 2004 due to decompensated heart failure. Only patients who had a functional capacity of class IV and who already received loop diuretic and ACE inhibitor therapy before their admission were included. Patients with recurrent admissions during this period were excluded. Eighty-five patients fulfilled these criteria: group I consisted of 25 patients who died during hospitalization whereas group II consisted of 60 patients who were discharged alive after treatment. Age, sex, left ventricular ejection fraction derived from 2-D echocardiography, serum sodium (Na), gamma-glutamyl transpeptidase (GGT), creatinine, uric acid levels, white blood cell counts and drugs used on admission were the selected parameters as predictors of in-hospital mortality in these patients. When stepwise logistic regression analysis was used, female sex and serum uric acid levels at admission appear to be the only predictors of death during that hospitalization independent of other variables. CONCLUSIONS: Serum uric acid levels may be used as a predictor of death in hospitalized heart failure patients with class IV symptoms. PMID- 16261780 TI - Diverse effects of calcium channel blockers in the collar model. AB - OBJECTIVE: Calcium channel blockers (CCBs) are among the most frequently prescribed cardiovascular drugs. It has been shown that these drugs have antiatherosclerotic effects in both experimental and clinical settings. However, calcium channel blockers have markedly different chemical structures and different effects on the cardiovascular system. We investigated the effect of CD 832, a Ca(+2) channel antagonist, on collar-induced intimal thickening, as well as accompanied reactivity changes in rabbit carotid artery. METHODS AND RESULTS: Rabbits received 5 mg/kg/day CD-832 or vehicle (polyethylene glycol, 0.5 ml/kg/day) intramuscularly for a week before and 2 weeks after the collar application. Histological and isometric force measurements were performed in segments from sham and collared carotid arteries. A three-week treatment with CD 832 did not inhibit the intimal thickening caused by perivascular application of a silicone collar. Potassium chloride (KCl), phenylephrine, 5-hydroxytryptamine (5-HT, serotonin) and histamine induced concentration-dependent contractions in both sham-operated (sham) and collared arteries. Collar-induced attenuations in maximum KCl, histamine, phenylephrine and 5-HT contractions were not affected by CD-832. Collaring caused an increase in pD2 values of 5-HT and a decrease in those of phenylephrine, histamine and acetylcholine. CD-832 did not affect the altered sensitivity to these agonists. CONCLUSIONS: These results demonstrate that, in rabbit carotid artery, CD-832 did not inhibit the collar-induced intimal thickening and did not affect the accompanying changes in vascular reactivity. PMID- 16261781 TI - The endothelial leukocyte adhesion molecule. Role in coronary artery disease. AB - OBJECTIVES: It is hypothesized that adhesion molecules could be an early predictor of coronary artery disease. The endothelial leucocyte adhesion molecule (ELAM) is expressed on activated endothelial cells only and it has been found to exist in a soluble form. This soluble form (sELAM) may be an important marker for endothelial cell damage. The aim of the present study was to compare the sELAM levels in coronary artery disease (CAD) subjects and healthy controls and to evaluate their clinical usefulness. METHODS AND RESULTS: sELAM were measured using enzyme immunoassay methods in 145 subjects having angiographically determined CAD and compared with 70 healthy, normotensive controls having a normal stress test/angiogram. Significantly higher values (p < 0.0001 ) were observed in CAD subjects as compared to controls. Also, subjects who underwent angioplasty and were later on readmitted with restenosis within 1 year had significantly higher levels of sELAM as compared to those who did not get restenosis within a year. CONCLUSIONS: These findings show that sELAM concentration is elevated in the presence of CAD and is useful for determining the presence of coronary atherosclerosis. An increased level of sELAM in patients susceptible to restenosis supports a role for white blood cell/endothelial interaction in restenosis after angioplasty. PMID- 16261783 TI - Percutaneous closure of a patent foramen ovale: single-centre experience using different types of devices and mid-term outcome. AB - INTRODUCTION: Patent foramen ovale (PFO) has been identified as a source of paradoxical embolism and cryptogenic stroke. Low recurrence rates of ischaemic stroke after percutaneous closure have been described. We report our single centre experience using four different types of devices. METHODS: All patients, who underwent a percutaneous PFO closure in the University Hospitals Leuven between February 1999 and December 2003, were included. The primary end point was defined as reoccurrence of stroke, transient ischaemic attack (TIA), or a peripheral thrombo-embolic event. The periprocedural and mid-term complications were reported. RESULTS: One-hundred and twelve patients, mean age 52.1 +/- 12.5 years (63 men/49 women), were included in the study. Indications for closure were cryptogenic stroke (91.9%), peripheral embolism (4.5%), obligate right-to-left shunt in Ebstein anomaly (1.8%), platypnoea syndrome or brain abscesses (both 0.9%). The Cardioseal/Starflex was used in 12, the Amplatzer PFO occluder in 35, the PFO Star/CardiaStar in 64, and the Helex in one patient. The primary end point occurred in 1.8% for stroke and 2.8% for TIA during a median follow-up of 1.9 years, range 4.9 years. Periprocedural complications were dislocation of the device (0.9%), transient arrhythmias (15.5%), aspiration pneumonia (0.9%), inguinal haematoma (3.6%), and an allergic reaction to medication (1.8%). Mid term complications were perforation of the device (0.9%), persistent transient arrhythmias (6.3%) and thrombus formation on the device (0.9%). No significant differences in outcome or the occurrence of any type of complication could be documented between the different types of devices. CONCLUSION: Percutaneous PFO closure seems to be a highly efficient and relatively safe procedure, independent of the type of device used for closure. PMID- 16261782 TI - N-acetyl-beta-D-glucosaminidase--a marker of reperfusion and a prognostic indicator in patients with acute myocardial infarction. AB - OBJECTIVE: N-acetyl-beta-D-glucosaminidase (NAG) is a lysosomal enzyme of which the activity in plasma is increased in a number of conditions including myocardial infarction. Plasma levels of cardiac proteins, such as myoglobin, troponin and creatine kinase, have been used as markers of myocardial reperfusion as well as for the prognosis of the disease. The aim of this study is to investigate whether NAG could be used as an additional biochemical marker to predict myocardial reperfusion and to find out if its release following thrombolysis has a prognostic value as well. METHOD: Streptokinase (SK) in a dose of 1.5 million units was administered intravenously to 75 patients with acute myocardial infarction (AMI) and the response to SK was assessed over a period of 90 minutes. Plasma NAG activity was monitored before (0 min) and 90 min after SK treatment. RESULTS: The mean NAG activity values were found to be 8.6 +/- 4.8 U/I and 10.95 +/- 7.3 U/I, respectively, and when compared using paired samples t test revealed a significant increase (p = 0.0001) following thrombolytic therapy. The increase appears perfusion related as rabbits injected with SK failed to show any increase in plasma NAG activity. There was an association between plasma NAG activity and clinical response to SK treatment (p = 0.045). A follow-up of 66 patients over a period of 18 months, revealed increased survival in AMI patients having significantly more activity/release of plasma NAG after thrombolytic treatment (p = 0.001). CONCLUSION: NAG appears to be another potential biochemical marker of reperfusion. Moreover, NAG release profile during thrombolysis could be of value in predicting prognosis of the disease. PMID- 16261784 TI - Forty-year mortality from cardiovascular diseases and their risk factors in men of the Italian rural areas of the Seven Countries Study. AB - OBJECTIVE: Forty-year cardiovascular mortality and its association with entry risk factor levels are reported for men enrolled in the Italian Rural Areas of the Seven Countries Study of Cardiovascular Diseases. METHODS AND RESULTS: Cardiovascular risk factors were measured in 1712 men aged 40-59 years at entry examination in 1960. Mortality data were collected during 40 years of follow-up. Overall death rate was 83.7%. The main causes of death were cardiovascular diseases, with a preponderance of those of atherosclerotic origin (CVDA, 33.7 % of all causes) including coronary heart disease, stroke and peripheral arterial diseases. Using 14 risk factors measured at baseline, multivariate analysis for CVDA showed that seven of them were strongly and significantly associated with events. The multivariate hazard ratio for 5 years of age was 1.59 (CI 1.43-1.77); for heavy job-related physical activity it was 0.96 (CI 0.79-1.18); for 10 cigarettes smoked per day 1.16 (CI 1.05-1.28); for 20 mm Hg of systolic blood pressure 1.38 (CI 1.25-1.52); for 1 mmol/l of serum cholesterol 1.19 (CI 1.09 1.30); for 0.25 l/m2 of height of vital capacity 0.87 (CI 0.78-0.97); for the presence of corneal arcus 1.36 (CI 1.03-1.79). Risk factors of which the coefficient did not reach statistical significance were family history for cardiovascular diseases, forced expiratory volume, heart rate, mid-arm circumference, subscapular skinfold, body mass index and diabetes. Similar findings were obtained for CHD and partially for stroke mortality. CONCLUSIONS: During a 40-year period classical cardiovascular risk factors were still highly predictive of cardiovascular diseases of atherosclerotic origin in these Italian rural populations. PMID- 16261785 TI - Relationship between low levels of high-density lipoprotein cholesterol and metabolic syndrome in Turkish patients. AB - OBJECTIVE: It is stated that high-density lipoprotein cholesterol (HDL-C) levels are low in Turkish people and that this may be related to genetic factors. Low HDL-C is a parameter of the metabolic syndrome (MetS). In this study, the relationship between low HDL-C levels and MetS has been investigated. METHODS AND RESULTS: A total of 720 successive patients, 20 years or older, were categorized into two groups, as those with low-HDL-C (in men < 40 mg/dl, in women < 50 mg/dl) and those without low-HDL-C levels (in men > or = 40 mg/dl, in women > or = 50 mg/dl). The groups were compared according to the frequency of the MetS and the averages of the MetS criteria other than HDL-C. Moreover, two groups were formed with normal (< 150 mg/dl) and high (> or = 150 mg/dl) triglyceride levels and they have been compared with the average HDL-C levels. The frequency of MetS was 70.8% in the group with low-HDL-C and 24.2% in the group without low HDL-C levels (p < 0.001). The averages of the MetS criteria other than the HDL-C were 2.17 +/- 1.19 in the group with low-HDL-C levels and 1.59 +/- 1.15 in the group without low-HDL-C levels (p < 0.001). Average HDL-C level was 48.90 +/- 13.40 mg/dl in the group with normal triglyceride levels and 44.41 +/- 10.26 mg/dl in the group with high triglyceride levels (p < 0.001). CONCLUSIONS: This study shows that low HDL-C is related to the MetS in our patients, a selected group of a population reported to have a low HDL-C average. PMID- 16261787 TI - Right atrial rhabdomyoma acting as the substrate for Wolff-Parkinson-White syndrome in a 3-month-old infant. AB - We report a 3-month-old infant girl who presented with supraventricular tachycardia, who was detected to have a large right atrial rhabdomyoma and evidence of Wolff-Parkinson-White (WPW) syndrome with an accessory pathway corresponding to the position of the tumour. In view of the resistant cardiac arrhythmia and large size of the tumour, the infant underwent surgical excision of the tumour. The evidence of WPW syndrome disappeared from the surface electrocardiogram immediately upon removal of the tumour. A second resolving tumour in the interventricular septum was also detected during the investigation. Although there are no other stigmata of tuberous sclerosis at present, our infant requires close follow-up. Right atrial rhabdomyomas are rare and to our knowledge this is the first reported instance where we could document clearly that the tumour itself formed the substrate for the WPW syndrome. PMID- 16261786 TI - Prognostic value of B-type natriuretic peptide levels on admission in patients with acute ST elevation myocardial infarction. AB - OBJECTIVE: To assess the relation between B-type natriuretic peptide (BNP) levels on admission in ST elevation myocardial infarction (STEMI) and short-term, all cause mortality. METHODS AND RESULTS: Blood samples for BNP determination were obtained on admission in 88 patients (mean age 60.6 +/- 10.7 years old) with STEMI. In a 15-minute period, BNP was measured by using simple bedside test for rapid quantification of BNP. Thirty days follow-up was performed. During the period of follow-up 12 (13.6%) patients died. Mean BNP was 228.74 +/- 269.98 pg/ml. The lowest value was 5 pg/ml, the highest value 1300 pg/ml due to limitations of the method. The baseline level of BNP was higher among patients who died than among those who were alive at 30 days (mean, 545.6 vs. 178.7 pg/ml; P = 0.001). Mortality increased among patients in increasing quartiles (p = 0.009). The unadjusted odds ratio for 30-day risk of death in the fourth quartile was 5.6 (95 percent confidence interval, 1.6 to 20.5; P < 0.001). When BNP was added to a multivariate Cox regression model including clinical and electrocardiographic variables, BNP levels were independently associated with the prognosis. CONCLUSIONS: BNP levels obtained on admission are a powerful, independent indicator of short-term mortality in patients with STEMI. Rapid tests for BNP assay seem to be a new tool in risk stratification of patients with STEMI. PMID- 16261788 TI - Arrhythmogenic right ventricular dysplasia presenting with severe right heart failure leading to abdominal ascites in a child. AB - This report describes a 12-year-old girl with arrhythmogenic right ventricular dysplasia, presenting with severe right ventricular failure symptoms including fatigue, abdominal pain and distension due to abdominal ascites. PMID- 16261790 TI - Preoperative use of intra-aortic balloon counterpulsation in very high-risk patients prior to urgent noncardiac surgery. AB - The optimal management of high-risk patients requiring urgent or emergency noncardiac surgery when the options for further testing and evaluation are limited, is unclear. The focus of most of the literature, including the American College of Cardiology/American Heart Association guidelines, has been on patients undergoing elective as opposed to urgent surgery. A few reports have suggested the use of intra-aortic balloon counterpulsation as a strategy to help carry such patients through the procedure. We describe three such very high-risk patients who successfully underwent emergency surgery with prophylactic insertion of an intra-aortic balloon pump without any adverse cardiac events or device-related complications. Our experience, as well as that of others suggests that preoperative intra-aortic balloon counterpulsation is a useful (and safe) approach to help very high-risk patients undergo urgent and emergency noncardiac surgery. Randomized trials are needed to assess whether balloon counterpulsation offers an incremental benefit over aggressive preoperative beta blockade. PMID- 16261789 TI - Myocardial infarction as an uncommon clinical manifestation of intravascular large cell lymphoma. AB - Intravascular large cell lymphoma (IVL) is a very rare variant of non-Hodgkin's lymphoma presenting with puzzling clinical manifestations. There is a predilection for the central nervous system, but the tumour often affects also skin, lung, and kidneys while lymphadenopathy and hepatosplenomegaly are usually absent. Myocardial infarction due to IVL has not been reported so far. We here report on a 56-year-old patient who was admitted to our hospital with fever and clinical signs of erysipelas. He had a 6-month history of "collagen vasculitic disease" treated with prednisolone and azathioprine. He received antibiotic treatment, but after transient improvement fever recurred with generalized seizures and myocardial infarction, which required transfer to the intensive care unit where the patient died with signs of an acute cardiogenic shock. Autopsy revealed a generalized high-grade B cell lymphoma of IVL type affecting and obstructing small vessels of a variety of tissues including heart, brain and lungs. The tumorous obliteration of small intramyocardial vessels had led to an acute ischaemia with infarction and subsequent signs of myocardial insufficiency. To the best of the authors' knowledge myocardial infarction as a leading symptom of IVL has not been described. PMID- 16261791 TI - Patient communication as a reflection of treatment philosophy. PMID- 16261792 TI - Principles and mechanisms of peri-implant soft tissue healing. AB - The long-term clinical and esthetic success of implant-supported restorations is determined by osseointegration and optimal remodeling of peri-implant soft tissues. Complications of soft-tissue management are often caused by fibrotic regeneration of oral mucosa after multiple surgical procedures. Knowledge of the proliferative processes in wound healing is necessary to attain adequate soft tissue conditions. Successful reconstruction of peri-implant soft tissues is feasible even in fibrotic conditions when appropriate surgical techniques are selected. The pleiotropic proliferative cytokine TGF-beta is involved in the regulation of all phases of wound healing and tissue remodeling. The isoform TGF beta1 is a cytokine associated with the development of fibrotic tissue. Overexpression of TGF-beta1 causes scarring and fibrosis, and results in limited clinical success of intraoral soft-tissue management. Experimental therapeutic approaches with neutralizing antibodies to block TGF-beta1 resulted in less scarring and a reduction of fibrosis. Further molecular biologic research of cell matrix-cytokine interactions in wound healing will provide highly specific antifibrotic therapeutic approaches in the future. PMID- 16261793 TI - Degree of conversion and surface hardness of resin cement cured with different curing units. AB - OBJECTIVE: The aim of this study was to evaluate the degree of conversion and Vickers surface hardness of resin cement under a simulated ceramic restoration with 3 different curing units: a conventional halogen unit, a high-intensity halogen unit, and a light-emitting diode system. METHODS AND MATERIALS: A conventional halogen curing unit (Hilux 550) (40 s), a high-intensity halogen curing unit used in conventional and ramp mode (Optilux 501) (10 s and 20 s, respectively), and a light-emitting diode system (Elipar FreeLight) (20 s, 40 s) were used in this study. The dual-curing resin cement (Variolink II) was cured under a simulated ceramic restoration (diameter 5 mm, height 2 mm), and the degree of conversion and Vickers surface hardness were measured. For degree of conversion measurement, 10 specimens were prepared for each group. The absorbance peaks were recorded using the diffuse-reflection mode of Fourier transformation infrared spectroscopy. For Vickers surface hardness measurement, 10 specimens were prepared for each group. A load of 200 N was applied for 15 seconds, and 3 evaluations of each of the samples were performed. RESULTS: Degree of conversion achieved with Optilux 501 (20 s) was significantly higher than those of Hilux, Optilux 501 (10 s), Elipar FreeLight (20 s), and Elipar FreeLight (40 s). For Vickers surface hardness measurement, Optilux 501 (20 s) produced the highest surface hardness value. No significant differences were found among the Hilux, Optilux 501 (10 s), Elipar FreeLight (20 s), and Elipar FreeLight (40 s). CONCLUSION: The high-intensity halogen curing unit used in ramp mode (20 s) produced harder resin cement surfaces than did the conventional halogen curing unit, high-intensity halogen curing unit used in conventional mode (10 s) and light-emitting diode system (20 s, 40 s), when cured through a simulated ceramic restoration. PMID- 16261794 TI - The patient with diabetes mellitus: etiology, epidemiology, principles of medical management, oral disease burden, and principles of dental management. AB - Diabetes mellitus is an etiologically and clinically heterogeneous group of metabolic disorders that share the commonality of hyperglycemia. Long-term hyperglycemia produces tissue damage, which ultimately manifests as microvascular and macrovascular disease, and neuropathy. The presence of macrovascular disease should alert clinicians to the possibility that the patient may have coronary artery disease, particularly because coronary artery disease and myocardial ischemia are likely to be silent. Elderly patients with diabetes mellitus are also more likely to develop congestive heart failure. Patients with unstable coronary syndromes, decompensated heart failure, and symptomatic cardiac arrhythmias are at increased risk of perioperative cardiovascular complications (myocardial infarction, heart failure, and sudden death) while undergoing noncardiac procedures. In addition, clinicians must avoid the risk of hypoglycemic episodes. Oral health care providers can expect to be called upon to care for patients with this progressively debilitating disease. To provide competent care to patients with diabetes mellitus, dental clinicians must understand the disease, its treatment, and the impact the disease and its treatment may have on the patient's ability to undergo and respond to dental care. PMID- 16261795 TI - Long-term evolution of a case of direct pulp capping by adhesion to dentin. AB - This article presents the long-term follow-up of a female patient who suffered pulp exposure during removal of a large caries lesion from the mandibular right first molar. The clinical decision to perform direct pulp capping was made. The tooth was treated with 5% chlorhexidine, etched with 37% phosphoric acid, and then restored with Heliomolar composite after application of Gluma dentin adhesive. The patient was followed for more than 8 years. PMID- 16261796 TI - Action of antimicrobial agents on infected odontogenic cysts. AB - OBJECTIVE: The treatment of choice for odontogenic cysts is surgery, and when the cysts are infected, preoperative antibiotic coverage is needed. However, the diffusion of antibiotics is a matter of controversy because of the low vascularization of the cystic epithelium. The aim of the present study was to determine the antimicrobial action of amoxicillin and metronidazole on infected odontogenic cysts. METHODS AND MATERIALS: Ten odontogenic root cysts were punctured before and after antibiotic treatment in 2 patient groups. Group 1 consisted of 5 patients treated with 500 mg amoxicillin at 6-hour intervals, and group 2 consisted of 5 patients treated with 400 mg metronidazole at 8-hour intervals, for 7 days. After this period, the patients were submitted to surgery for enucleation. The fluid collected was seeded onto culture media for counts of total bacteria and facultative anaerobic bacteria. RESULTS: The results showed that the amount of bacteria present in the fluid was significantly higher than the amount isolated after antibiotic treatment. Also, most of the microorganisms present in the fluid collected before antibiotic administration were strict anaerobes. CONCLUSIONS: Both antibiotics reduced the number of bacteria in the cystic fluid, showing that they did diffuse into the lesions at sufficient concentrations to exert their antimicrobial action. PMID- 16261797 TI - Antimicrobial spray for toothbrush disinfection: an in vivo evaluation. AB - OBJECTIVE: The aim of this study was to evaluate the efficacy of a spray containing an antimicrobial solution for toothbrush disinfection. METHODS AND MATERIALS: Three different solutions were sprayed on toothbrush bristles among 30 adults after they had brushed: (1) basic formulation (base) plus chlorhexidine; (2) base only, and (3) sterile tap water (control). Each solution was tested for 1 week. After that, the toothbrushes were collected and sonicated in Letheen Broth, diluted in 10-fold series, and plated on selective and nonselective media for detection of anaerobes, aerobes, streptococci, and gram-negative bacilli. After incubation, the colonies of those microorganisms were counted. Presence of mutans streptococci on the bristles was also confirmed. RESULTS: Spray 1 produced a significant reduction in the microbial contamination of toothbrushes for all the microorganisms, spray 2 provided some reduction of contaminants, and spray 3 demonstrated the least antimicrobial effect. CONCLUSION: The antimicrobial spray with chlorhexidine proved to be an effective and practical means for toothbrush disinfection. PMID- 16261800 TI - Clinical images in oral medicine. Angioedema. PMID- 16261798 TI - Effects of adhesive liner and provisional cement on the bond strength of nickel/chrome/beryllium alloy cemented to dentin. AB - OBJECTIVE: Treating teeth with adhesive agents before placing a provisional restoration can prevent tooth sensitivity. This study evaluated the bond strength of resin cements to dentin treated with 2 adhesive agents and 2 provisional cements. METHODS AND MATERIALS: Extracted human molars were prepared by exposing dentin and were treated with either Prime & Bond NT or Clearfil SE Bond. After a simulated impression technique, the teeth were provisionalized with either a eugenol or noneugenol temporary cement. Teeth were cleaned for bonding by either mechanical removal of the cement or use of an acid conditioner. Panavia F and Calibra resin cements were used to cement nickel/chrome/beryllium alloy to the tooth surfaces, and the specimens were debonded. Mean shear bond strengths for each group were calculated. RESULTS: Mean shear bond strengths ranged from 26.6 +/- 5.8 MPa for Calibra bonded to dentin treated with Prime & Bond NT, a noneugenol cement, and mechanically cleaned, to 10.6 +/- 4.4 MPa for Panavia F bonded to unlined (no adhesive) dentin treated with a eugenol cement and mechanically cleaned. Of the 14 groups tested, significant differences were observed related to the adhesives and resin cements. Both temporary cements reduced the bond to dentin not treated with a resin adhesive. Use of an acid conditioner for cleaning the temporary cement also reduced bond strengths in all groups. CONCLUSIONS: Placement of a dentin adhesive before provisionalization may prevent the temporary cement from affecting the bond of the final resin cement to the tooth. For the products used in this study, use of phosphoric acid to clean the tooth surface is not recommended. PMID- 16261799 TI - Study of denture-induced fibrous hyperplasia cases diagnosed from 1979 to 2001. AB - OBJECTIVE: The purpose of this research was to study the cases of inflammatory fibrous hyperplasia (IFH) at the Clinic of Semiology, Department of Bioscience and Oral Diagnosis, Sao Jose dos Campos Dental School, State University of Sao Paulo, Brazil. METHODS AND MATERIALS: A total of 141 clinical file cards indicating a final diagnosis of IFH, from the archives of the Department of Bioscience and Oral Diagnosis and dated from 1979 to 2001, were included in the study. Of these files, 50 indicated a diagnosis of denture-induced fibrous hyperplasia. Sex, age, race, duration, and clinical features that confirm their classification in the non-neoplastic proliferating process were analyzed statistically. RESULTS: Of the 50 analyzed cases of denture-induced lesion, 22% occurred in men and 78% in women. Patients in the age group of 41 to 50 years presented the highest frequency of the lesion. CONCLUSIONS: Inflammatory fibrous hyperplasia occurs more frequently in women (71.63%), and denture-induced lesions appear mainly in patients over 40 years of age (70% of cases). Patients with denture-induced hyperplasia reported pain associated with the lesion (70%). PMID- 16261802 TI - A reappraisal of the ascending systems in man, with emphasis on the medial lemniscus. PMID- 16261801 TI - Chronic osteomyelitis of the mandible. PMID- 16261803 TI - Axonal branching and recovery of coordinated muscle activity after transection of the facial nerve in adult rats. AB - Facial nerve surgery inevitablyleads to pareses, abnormally associated movements, and pathologically altered reflexes. The reason for this "post-paralytic syndrome" is the misdirected reinnervation of targets, which consists of two major components. First, due to malfunctioning axonal guidance, a muscle gets reinnervated by a "foreign" axon, that has been misrouted along a "wrong" fascicle. Second, the supernumerary collateral branches emerging from all transected axons simultaneously innervate antagonistic muscles and cause severe impairment of coordinated activity. Since it is hardly possible to influence the first major component and improve the guidance of several thousands of axons, we concentrated on the second major component and tried to reduce the collateral axonal branching. The efficiency of various treatments was evaluated in rats by determining: (1) the degree of post-operative axonal branching as estimated by the number of double-or triple-labeled perikarya after application of crystalline DiI, Fluoro-Gold (FG), and Fast Blue (FB) to the zygomatic, buccal, and marginal mandibular branch of the facial nerve respectively; (2) the accuracy of reinnervation as estimated by the number of double-labeled perikarya innervating the whisker pad muscles before and after surgery as shown by intramuscular injections of FG and FB respectively; (3) the recovery of vibrissal motor performance, estimated by a video based motion analysis. So far, we have tried to reduce branching by alteration of the afferent trigeminal input to the axotomized facial motoneurons and by focal application of: (1) neurite outgrowth fostering ECM proteins; (2) neutralizing antibodies to NGF, BDNF, CNTF, GDNF, IGF-I, and FGF-II; (3) suspensions of olfactory ensheathing cells, Schwann cells, and bone marrow stroma cells; and (4) pieces of autologous olfactory mucosa to the transection site. Although most of these manipulations do influence peripheral nerve regeneration to some extent, only the application of autologous olfactory mucosa yielded a major improvement, i.e., better function. PMID- 16261804 TI - Aquaculture of "non-food organisms" for natural substance production. AB - Marine invertebrates are already sources of commercially important secondary metabolites and may become even more so as knowledge on marine natural products and chemical ecology develops. Among the producers of these compounds predominantly sponges, bryozoa and molluscs have received the attention of academic and industrial research and development. For all these invertebrate groups culture techniques have been developed encompassing in situ, laboratory and cell culture approaches for the production of natural products. Potential applications of these are not restricted to pharmaceuticals but include marine cements, biominerals and antifouling compounds. In addition, markets exist for ornamental species. All culture approaches require sound ecological knowledge about the organisms to be cultured and possible symbiotic interactions between host invertebrates and microheterotrophs. PMID- 16261805 TI - Bioprocess engineering data on the cultivation of marine prokaryotes and fungi. AB - The temperature/pressure dependency of marine prokaryotes and fungi, in terms of their growth behaviour as well as their potential to produce new metabolites or enzymes, is evaluated. Advanced shake-flask cultivations and controlled bioreactor cultivations following the batch-type, fed-batch-type and/or continuous-type procedures are summarized. After a summary of the fermentation data available so far, values on maximal biomass, specific growth rates, and (sub)optimal production yields are presented. The application of mesophilic microbes, especially bioactive metabolites, to intensify bioprocess engineering studies, is the goal. Cold-active enzymes and thermostable enzymes are the targets of experiments with psychrophilic and hyperthermophilic enzymes. A special challenge to bioengineers is also provided by barophilic strains originating from depths of, say, nearly 11000 m, or from hydrothermal vents. PMID- 16261806 TI - Downstream processing in marine biotechnology. AB - Downstream processing is one of the most underestimated steps in bioprocesses and this is not only the case in marine biotechnology. However, it is well known, especially in the pharmaceutical industry, that downstreaming is the most expensive and unfortunately the most ineffective part of a bioprocess. Thus, one might assume that new developments are widely described in the literature. Unfortunately this is not the case. Only a few working groups focus on new and more effective procedures to separate products from marine organisms. A major characteristic of marine biotechnology is the wide variety of products. Due to this variety a broad spectrum of separation techniques must be applied. In this chapter we will give an overview of existing general techniques for downstream processing which are suitable for marine bioprocesses, with some examples focussing on special products such as proteins (enzymes), polysaccharides, polyunsaturated fatty acids and other low molecular weight products. The application of a new membrane adsorber is described as well as the use of solvent extraction in marine biotechnology. PMID- 16261807 TI - Marine pharmacology: potentialities in the treatment of infectious diseases, osteoporosis and Alzheimer's disease. AB - Several molecules isolated from various marine organisms (microorganisms, algae, fungi, invertebrates, and vertebrates) are currently under study at an advanced stage of clinical trials, either directly or in the form of analogues deduced from structure-activity relationships. Some of them have already been marketed as drugs. The goal of this article is not to present a complete panorama of marine pharmacology but to show that new models and new mechanisms of action of marine substances bring new solutions for tackling some of the major public health problems of the 21st century. These include: malaria, which assails mainly the southern hemisphere; tuberculosis, an infectious disease once believed to be eliminated but alarmingly increasing, especially among HIV-positive populations; and osteoporosis and Alzheimer's disease, the extension of which are correlated with ageing populations, especially in the developed countries. PMID- 16261808 TI - Asymmetric total synthesis of complex marine natural products. AB - Among nature's ecosystems, the marine environment has been an extremely rich source of structurally complex and biologically active molecules. This review aims to cover the recent developments in the synthesis of marine natural products, also reflecting the trend of their increased use to address biological questions. The examples chosen should be viewed as representative of the different structural motifs on the one hand and the strategies and stimuli for their synthesis on the other. PMID- 16261809 TI - Seafood allergy: lessons from clinical symptoms, immunological mechanisms and molecular biology. AB - Food allergy consists of a wide range of disorders that result from adverse immune responses to dietary antigens. Manifestations of allergic response includes acute, potentially fatal anaphylactic reactions and a variety of chronic diseases that mainly affect the gastrointestinal tract, skin, and respiratory tract. Tools for clinical diagnosis and management, which have not changed much in the past two decades, include the clinical history, tests for specific IgE antibody to suspected foods, elimination diets, oral food challenges, and provision of medications such as epinephrine for emergency treatment. On the other hand, recent immunological and molecular biological research have enhanced our understanding of the mechanisms of these disorders and revealed the identities of many food allergens. Here, we will discuss seafood allergies with respect to the clinical manifestations, diagnosis, immunological mechanisms, and molecular biology of seafood allergens. Furthermore, potential applications and future directions in the clinical management of seafood allergies are discussed. PMID- 16261810 TI - Nursing research committee makes plans for the coming year. AB - Evidence-based practice (EBP) enhances the care of patients and families by guiding nurses in their patient care. Nursing leaders, health care administrators and policy-makers are increasingly requiring that nursing practice be based on the best available evidence. Murdoch (2004) stated that a review and synthesis of existing research is essential to identify the best available evidence. Koop (2002) further reflected on the challenges faced by oncology nurses when there are clinical situations for which there is little or no empirical evidence on which to base nursing decisions. A need to learn skills and gain confidence in identifying the best available evidence frequently motivates staff nurses and clinicians to learn more about critical appraisal of the research literature. At the McGill University Health Centre (MUHC) these needs have prompted nurses to enrol in the research utilization (RU) course that the health centre offers. This column will outline the context and components of the RU course and summarize feedback from participants. Challenges and limitations of this strategy and its relationship to oncology nursing practice are highlighted. PMID- 16261811 TI - Needs of patients and families undergoing autologous peripheral blood stem cell transplantation. AB - Autologous peripheral blood stem cell transplantation (APBSCT) as a rescue for myeloablative chemotherapy has become the standard of care for several malignancies. The majority of pre-transplant treatment is provided in outpatient oncology clinics and early discharge post-transplant is facilitated by the use of colony stimulating factors. The patient/family is then required to manage complex self-care in the home environment (Johns, 1998; Poloquin, 1997; Schulmeister et al., 2005). This needs assessment was undertaken to determine how to improve the quality of care delivery for patients/families undergoing APBSCT. This qualitative research utilized unstructured interviews, observation, and field notes for data collection. Data were analyzed using thematic content analysis. Four key concepts to enhance the quality of care delivery emerged: knowledge, communication, support, and skill. PMID- 16261812 TI - Peripheral nerve function and symptom distress during biotherapy for malignant melanoma. AB - Studies of biotherapy-induced physiological changes are few, and systematic monitoring for neurotoxic effects are lacking. The purpose of this exploratory pilot study was to determine the change in peripheral nerve function and symptom distress during treatment with biotherapy for malignant melanoma. A convenience sample of 11 participants with malignant melanoma receiving interferon-alpha had measures of peripheral nerve function measured at baseline, four and 12 weeks of treatment. Data were analyzed using plots and regression slopes to determine change over time in sensation, gait/balance, vision, hearing, vibratory sense, muscle strength, deep tendon reflexes, blood pressure, and symptom distress. Declines in hearing, sensation, vibration, and muscle strength were found. Changes in visual acuity, and orthostatic blood pressure were noted, while gait/balance remained stable. Additionally, neuropathy symptoms were associated with symptom distress. The characterization of such changes can increase our understanding of the nature of the physiological effects associated with high dose biotherapy treatment and aid clinicians to better prepare patients for anticipated changes in function and subsequent lifestyle adjustments. These findings can be used to provide information in a larger study of this phenomenon regarding important outcomes and measurement time-points of therapy-induced neuropathy and decreasing symptom distress in patients receiving cancer treatment with biologic agents. PMID- 16261814 TI - Advocacy: the role of oncology nursing. PMID- 16261813 TI - Telling the story of SARS: compassionate oncology care in the face of a futuristic health crisis. PMID- 16261815 TI - Palliative nurses' interest group--June 2005 report. PMID- 16261816 TI - A chemotherapy, hypersensitivity and anaphylaxis protocol. PMID- 16261817 TI - When too much is still not enough. PMID- 16261818 TI - Expression of inhibitory G proteins in adenomatous thyroid glands obtained from hyperthyroid cats. AB - OBJECTIVE: To identify within guanosine triphosphate-binding proteins (G proteins) the subset of inhibitory G proteins (G) that have decreased expression in adenomatous thyroid glands obtained from hyperthyroid cats. SAMPLE POPULATION: Adenomatous thyroid glands obtained from 5 hyperthyroid cats and normal thyroid glands obtained from 3 age-matched euthyroid cats. PROCEDURE: Expression of G(i1), G(i2), and G(i3) in enriched membrane preparations from thyroid glands was quantified by use of immunoblotting with G(i) subtype-specific antibodies. RESULTS: Expression of G(i2) was significantly decreased in tissues of hyperthyroid glands, compared with expression in normal thyroid tissue. Expression of G(i1) and G(i3) was not significantly different between normal thyroid tissues and tissues from hyperthyroid glands. CONCLUSIONS AND CLINICAL RELEVANCE: A decrease in G(i2) expression decreases inhibition of adenylyl cyclase and allows a relative increase in stimulatory G protein expression. This results in increased amounts of cAMP and subsequent unregulated mitogenesis and hormone production in hyperthyroid cells. Decreased G(i2) expression may explain excessive growth and function of the thyroid gland in cats with hyperthyroidism. PMID- 16261819 TI - Myocardial concentrations of fatty acids in dogs with dilated cardiomyopathy. AB - OBJECTIVE: To compare myocardial concentrations of fatty acids in dogs with dilated cardiomyopathy (DCM) with concentrations in control dogs. SAMPLE POPULATION: Myocardial tissues from 7 dogs with DCM and 16 control dogs. PROCEDURE: Myocardial tissues were homogenized, and total fatty acids were extracted and converted to methyl esters. Myocardial concentrations of fatty acids were analyzed by use of gas chromatography and reported as corrected percentages. RESULTS: The amount of docosatetraenoic acid (C22:4 n-6) was significantly higher in myocardial samples from dogs with DCM (range, 0.223% to 0.774%; median, 0.451%), compared with the amount in samples obtained from control dogs (range, 0.166% to 0.621%; median, 0.280%). There were no significant differences between DCM and control dogs for concentrations of any other myocardial fatty acids. CONCLUSIONS AND CLINICAL RELEVANCE: Although concentrations of most myocardial fatty acids did not differ significantly between dogs with DCM and control dogs, the concentration of docosatetraenoic acid was significantly higher in dogs with DCM. Additional investigation in a larger population is warranted to determine whether this is a primary or secondary effect of the underlying disease and whether alterations in fatty acids may be a target for intervention in dogs with DCM. PMID- 16261820 TI - Effects of zoledronate on markers of bone metabolism and subchondral bone mineral density in dogs with experimentally induced cruciate-deficient osteoarthritis. AB - OBJECTIVE: To evaluate effects of zoledronate on markers of bone metabolism in dogs after transection of the cranial cruciate ligament (CrCL). ANIMALS: 21 adult dogs. PROCEDURE: Unilateral CrCL transection was performed arthroscopically. Dogs were allocated to 3 groups (control group, low-dose zoledronate [10 microg/kg, SC, q 90 d for 12 months], and high-dose zoledronate [25 microg/kg, SC, q 90 d for 12 months]). Serum osteocalcin (OC), serum bone-specific alkaline phosphatase (BAP), and urine pyridinoline and deoxypyridinoline concentrations were measured at 0, 1, 3, 6, 9, and 12 months after surgery. Bone mineral density (BMD) was determined in the distal portion of the femur and proximal portion of the tibia via computed tomography at each time point. Data were analyzed by a repeated measures ANOVA. RESULTS: oledronate inhibited OC in the high-dose group at 9 and 12 months and at 12 months in the low-dose group, compared with the control group. High-dose zoledronate decreased BAP concentrations 3 and 9 months after surgery. In the control group, BMD was decreased in the femoral condyle and caudal tibial plateau. Zoledronate prevented significant BMD decreases starting 1 month after transection, compared with control dogs. In the caudomedial aspect of the tibial plateau, both zoledronate groups had significant increases in BMD after 3 months, compared with control dogs. CONCLUSIONS AND CLINICAL RELEVANCE: Zoledronate may reduce subchondral bone loss and effect markers of bone metabolism in dogs with experimentally induced instability of the stifle joint and subsequent development of osteoarthritis. PMID- 16261821 TI - Cardiopulmonary effects of dexmedetomidine in sevoflurane-anesthetized sheep with and without nitric oxide inhalation. AB - OBJECTIVE: To determine whether inhaled nitric oxide (NO) prevents pulmonary hypertension and improves oxygenation after i.v. administration of a bolus of dexmedetomidine in anesthetized sheep. ANIMALS: 6 healthy adult sheep. PROCEDURE: In a crossover study, sevoflurane-anesthetized sheep received dexmedetomidine (2 microg/kg, i.v.) without NO (DEX treatment) or with inhaled NO (DEX-NO treatment). Cardiopulmonary variables, including respiratory mechanics, were measured before and for 120 minutes after bolus injection of dexmedetomidine. RESULTS: Dexmedetomidine induced a transient decrease in heart rate and cardiac output. A short-lived increase in mean arterial pressure (MAP) and systemic vascular resistance (SVR) was followed by a significant decrease in MAP and SVR for 90 minutes. Mean pulmonary arterial pressure (MPAP) and pulmonary vascular resistance increased transiently after dexmedetomidine injection. The Pao2 was significantly decreased 3 minutes after injection and reached a minimum of (mean +/- SEM) 13.3 +/- 78 kPa 10 minutes after injection. The decrease in Pao2 was accompanied by a sudden and prolonged decrease in dynamic compliance and a significant increase in airway resistance, shunt fraction, and alveolar dead space. Peak changes in MPAP did not differ between the 2 treatments. For the DEX NO treatment, Pao2 was significantly lower and the shunt fraction significantly higher than for the DEX treatment. CONCLUSIONS AND CLINICAL RELEVANCE: Inhalation of NO did not prevent increases in pulmonary arterial pressures induced by i.v. administration of dexmedetomidine. Preemptive inhalation of NO intensified oxygenation impairment, probably through increases in intrapulmonary shunting. PMID- 16261822 TI - Effects of the omega-3 fatty acid, alpha-linolenic acid, on lipopolysaccharide challenged synovial explants from horses. AB - OBJECTIVE: To determine the effects of pretreatment with alpha-linolenic acid, an omega-3 polyunsaturated fatty acid, on equine synovial explants challenged with lipopolysaccharide (LPS). ANIMALS: 8 mature mixed-breed horses (4 mares and 4 geldings). PROCEDURE: Synovial explants were assigned to receive 1 of 7 concentrations of alpha-linolenic acid, ranging from 0 to 300 microg/mL. At each concentration, half of the explants were controls and half were challenged with 0.003 microg of LPS as a model of synovial inflammation. Cell inflammatory response was evaluated by measurement of prostaglandin E2 production via an ELISA. Synovial cell viability, function, histomorphologic characteristics, and cell membrane composition were evaluated by use of trypan blue dye exclusion, hexuronic acid assay for hyaluronic acid, objective microscopic scoring, and high performance liquid chromatography, respectively. RESULTS: Challenge with LPS significantly increased production of prostaglandin E2 and decreased production of hyaluronic acid. Treatment with alpha-linolenic acid at the highest dose inhibited prostaglandin E2 production. Cell viability and histomorphologic characteristics were not altered by treatment with alpha-linolenic acid or LPS challenge. Treatment with alpha-linolenic acid increased the percentage of this fatty acid in the explant cell membranes. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that investigation of alpha-linolenic acid as an anti inflammatory medication for equine synovitis is warranted. PMID- 16261823 TI - Concentrations of serum amyloid A and lipopolysaccharide-binding protein in horses with colic. AB - OBJECTIVE: To determine concentrations of 2 acute-phase proteins (serum amyloid A [SAA] and lipopolysaccharide-binding protein [LBP]) in serum samples obtained from horses with colic and identify relationships among these acute-phase proteins and clinical data. ANIMALS: 765 horses with naturally developing gastrointestinal tract diseases characterized by colic (ie, clinical signs indicative of abdominal pain) and 79 healthy control horses; all horses were examined at 2 university teaching hospitals. PROCEDURE: Serum concentrations of SAA and LBP were determined by immunoturbidometric and dot-blot assays, respectively. RESULTS: SAA and LBP concentrations were determined for 718 and 765 horses with colic, respectively. Concentrations of SAA were significantly higher in nonsurvivors than in survivors, and horses with enteritis or colitis and conditions characterized by chronic inflammation (eg, abdominal abscesses, peritonitis, or rectal tears) had SAA concentrations significantly greater than those for horses with other conditions. Serum concentrations of LBP did not correlate with outcome, disease process, or portion of the gastrointestinal tract affected. CONCLUSIONS AND CLINICAL RELEVANCE: Circulating concentrations of SAA were significantly higher at admission in horses with colic attributable to conditions having a primary inflammatory cause (eg, enteritis, colitis, peritonitis, or abdominal abscesses) and were higher in horses that failed to survive the episode of colic, compared with concentrations in horses that survived. Serum concentrations of LBP did not correlate with survival. Analysis of these findings suggests that evaluation of SAA concentrations may be of use in identifying horses with colic attributable to diseases that have inflammation as a primary component of pathogenesis. PMID- 16261824 TI - Retention of ingested porcine reproductive and respiratory syndrome virus in houseflies. AB - OBJECTIVE: To evaluate retention of porcine reproductive and respiratory syndrome virus (PRRSV) in houseflies for various time frames and temperatures. SAMPLE POPULATION: Fifteen 2-week-old pigs, two 10-week-old pigs, and laboratory cultivated houseflies. PROCEDURE: In an initial experiment, houseflies were exposed to PRRSV; housed at 15 degrees, 20 degrees, 25 degrees, and 30 degrees C; and tested at various time points. In a second experiment to determine dynamics of virus retention, houseflies were exposed to PRRSV and housed under controlled field conditions for 48 hours. Changes in the percentage of PRRSV-positive flies and virus load per fly were assessed over time, and detection of infective virus at 48 hours after exposure was measured. Finally, in a third experiment, virus loads were measured in houseflies allowed to feed on blood, oropharyngeal washings, and nasal washings obtained from experimentally infected pigs. RESULTS: In experiment 1, PRRSV retention in houseflies was proportional to temperature. In the second experiment, the percentage of PRRSV-positive houseflies and virus load per fly decreased over time; however, infective PRRSV was found in houseflies 48 hours after exposure. In experiment 3, PRRSV was detected in houseflies allowed to feed on all 3 porcine body fluids. CONCLUSIONS AND CLINICAL RELEVANCE: For the conditions of this study, houseflies did not support PRRSV replication. Therefore, retention of PRRSV in houseflies appears to be a function of initial virus load after ingestion and environmental temperature. These factors may impact the risk of insect-borne spread of PRRSV among farms. PMID- 16261826 TI - In vitro biomechanical comparison of a plate-rod combination-construct and an interlocking nail-construct for experimentally induced gap fractures in canine tibiae. AB - OBJECTIVE: To compare structural properties of a plate-rod combination-bone construct (PRCbc) and interlocking nail-bone construct (ILNbc) by use of an experimentally induced gap fracture in canine tibiae. SAMPLE POPULATION: 12 paired canine tibiae. PROCEDURE: Specimens were implanted with a plate-rod combination consisting of a 3.5-mm, limited-contact, dynamic-compression plate combined with an intramedullary rod or 6-mm interlocking nail. Ostectomy (removal of 10-mm segment) was performed. Paired constructs were loaded for bending, compression, or torsion measurements (4 constructs/group). Compliance was determined by fitting regression lines to the load-position curves at low (initial compliance) and high (terminal compliance) loads. RESULTS: Bending compliances did not differ significantly between constructs. For the ILNbc, initial compliance was greater than terminal compliance in compression and torsion. Initial compliance and terminal compliance for the PRCbc were similar in compression and torsion. Initial compliance in compression and torsion was greater for the ILNbc, compared with initial compliance for the PRCbc. Maximum deformations in bending and compression were similar between constructs; however, maximum torsional angle was significantly greater for the ILNbc, compared with values for the PRCbc. CONCLUSIONS AND CLINICAL RELEVANCE: The study documented that for an experimentally induced gap fracture in canine tibiae, a plate-rod combination is a significantly less compliant fixation method in torsion and compression, compared with an interlocking nail. Considering the deleterious effects of torsional deformation on bone healing, a plate-rod combination may represent a biomechanically superior fixation method, compared with an interlocking nail, for the treatment of dogs with comminuted tibial diaphyseal fractures. PMID- 16261825 TI - Isolation, characterization, and expression of stromelysin-1 in primary tumors of dogs. AB - OBJECTIVE: To isolate and characterize the cDNA sequence of canine stromelysin-1 (matrix metalloproteinase [MMPI-3), screen various naturally developing primary tumors of dogs, and assess the effect of stromelysin-1 on survival of dogs with cancer. SAMPLE POPULATION: 3 canine cell lines and biopsy specimens of primary tumors collected from 54 dogs. PROCEDURE: 3 canine cell lines and biopsy specimens of primary tumors collected from 54 dogs at the University of Illinois Veterinary Teaching Hospital were used in the study. Primer sets based on human stromelysin-1 and consensus sequences were designed for expression, screening, and isolation. Two additional primer sets were designed for screening. Samples were assayed at least in duplicate. Data were analyzed for differences in expression of stromelysin-1 on the basis of sex, age, metastasis, malignant versus nonmalignant tissue origin, and duration of patient survival. RESULTS: A 1,479-bp cDNA nucleotide sequence was amplified from established canine cell lines. The open reading frame encoded a protein consisting of 478 amino acids. This sequence was 70% to 88% homologous with stromelysin-1 of other species at the amino acid level. Fifty-four samples were screened for stromelysin-1. Of these, 34 (63%) had positive results and 20 (37%) had negative results for expression. Stromelysin-1 and metastasis were associated with a poor prognosis for survival. CONCLUSIONS AND CLINICAL RELEVANCE: Stromelysin-1 is a potential activator of other members of the MMP family. Additional studies are needed to investigate the relationship between stromelysin-1 production and aggressive biological behavior of tumors in dogs. PMID- 16261827 TI - Pulmonary arterial disease in cats seropositive for Dirofilaria immitis but lacking adult heartworms in the heart and lungs. AB - OBJECTIVE: To determine the prevalence and severity of pulmonary arterial lesions in cats seropositive for heartworms (Dirofilaria immitis) but lacking adult heartworms in the heart and lungs during necropsy. ANIMALS: 630 adult cats from an animal control shelter in Florida. PROCEDURE: Cats were tested for adult heartworms in the heart and pulmonary arteries and antibody against heartworms in the serum. Histologic examination was conducted on the right caudal lung lobe of 24 heartworm- and antibody-positive cats; 24 heartworm-negative and antibody positive cats; and 24 heartworm-, antibody-, and antigen-negative cats. Wall areas of 10 small to medium-sized pulmonary arteries of each cat were measured and expressed as a proportion of total cross-sectional area. RESULTS: Heartworm infection or seropositive status was significantly and strongly associated with seventy of medial hypertrophy of pulmonary arterial walls. Heartworm- and antibody-positive cats and heartworm-negative and antibody-positive cats had a significant increase in wall thickness, compared with wall thickness for heartworm- and antibody-negative cats. Heartworm- and antibody-positive cats had the most severe hypertrophy. The proportion with occlusive medial hypertrophy was significantly higher in heartworm- and antibody-positive cats (19/24 [79%]) and heartworm-negative and antibody-positive cats (12/24 [50%]), compared with heartworm- and antibody-negative cats (3/24 [13%]). CONCLUSIONS AND CLINICAL RELEVANCE: Cats with serologic evidence of exposure to heartworms, including those without adult heartworms in the lungs and heart, have a greater prevalence of pulmonary arterial lesions than heartworm-negative cats without serologic evidence of exposure. Additional studies are needed to define the pathogenesis, specificity, and clinical importance of these lesions. PMID- 16261828 TI - Relative sensitivity of polymerase chain reaction assays used for detection of feline herpesvirus type 1 DNA in clinical samples and commercial vaccines. AB - OBJECTIVE: To determine relative detection rates and detection limits for 6 published polymerase chain reaction (PCR) assays used for detection of feline herpesvirus type 1 (FHV-1) DNA. SAMPLE POPULATION: 5 vaccines licensed for use in preventing FHV-1-associated disease; 15 conjunctival biopsy specimens collected from cats with keratitis, conjunctivitis, or both; and a plaque-purified field isolate of FHV-1 cultured in vitro. PROCEDURE: Vaccines and clinical samples were assessed for FHV-1 DNA by use of all 6 assays. Detection rates were calculated by assuming that any sample in which FHV-1 DNA was detected was a true-positive result. Detection limits were estimated by use of serial dilutions of DNA extracted from cultured FHV-1 and 1 clinical sample. RESULTS: Testing by use of all 6 assays resulted in detection of FHV-1 DNA in all 5 vaccines. Testing by use of all 6 assays yielded concordant results for 9 of 15 conjunctival biopsy specimens (8 with negative results and 1 with a positive result). Calculated detection rates for clinical samples ranged from 29% to 86%. Assay sensitivity was ranked similarly by use of detection rate or detection limit. CONCLUSIONS AND CLINICAL RELEVANCE: Testing by use of all assays was equally likely to detect vaccine virus. Therefore, a positive PCR result in a cat may reflect vaccine virus rather than wild-type virus. Test sensitivity as assessed by detection limits and detection rates varied greatly. Because FHV-1 can be shed in clinically normal animals, high detection rate will not necessarily correlate with high diagnostic sensitivity. PMID- 16261829 TI - Assessment of the qualitative variation in bacterial microflora among compartments of the intestinal tract of dogs by use of a molecular fingerprinting technique. AB - OBJECTIVE: To evaluate the qualitative variation in bacterial microflora among compartments of the intestinal tract of dogs by use of a molecular fingerprinting technique. ANIMALS: 14 dogs (similarly housed and fed identical diets). PROCEDURE: Samples of intestinal contents were collected from the duodenum, jejunum, ileum, colon, and rectum of each dog. Bacterial DNA was extracted from the samples, and the variable V6 to V8 region of 16S ribosomal DNA (gene coding for 16S ribosomal RNA) was amplified by use of universal bacterial primers; polymerase chain reaction amplicons were separated via denaturing gradient gel electrophoresis (DGGE). Similarity indices of DGGE banding patterns were used to assess variation in the bacterial microflora among different compartments of the intestine within and among dogs. Bacterial diversity was assessed by calculating the Simpson diversity index, the Shannon-Weaver diversity index, and evenness. RESULTS: DGGE profiles indicated marked differences in bacterial composition of intestinal compartments among dogs (range of similarity, 25.6% to 36.6%) and considerable variation among compartments within individual dogs (range of similarity, 36.7% to 579%). Similarities between neighboring intestinal compartments were significantly greater than those between non-neighboring compartments. Diversity indices for the colon and rectum were significantly higher than those of the duodenum, jejunum, and ileum. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that the different intestinal compartments of individual dogs appear to host different bacterial populations, and these compartmental populations vary among dogs. In dogs, fecal sample analysis may not yield accurate information regarding the composition of bacterial populations in compartments of the gastrointestinal tract. PMID- 16261830 TI - Distribution of power across the hind limb joints in Labrador Retrievers and Greyhounds. AB - OBJECTIVE: To quantify angular excursions; net joint moments; and powers across the stifle, tarsal, and metatarsophalangeal (MTP) joints in Labrador Retrievers and Greyhounds and investigate differences in joint mechanics between these 2 breeds of dogs. ANIMALS: 12 clinically normal dogs (6 Greyhounds and 6 Labrador Retrievers) with no history of hind limb lameness. PROCEDURE: Small retroreflective markers were applied to the skin over the pelvic limb joints, and a 4-camera kinematic system captured data at 200 Hz in tandem with force platform data while the dogs trotted on a runway. Breed-specific morphometric data were combined with kinematic and force data in an inverse-dynamics solution for stance phase net joint moments and powers at the stifle, tarsal, and MTP joints. RESULTS: There were gross differences in kinematic patterns between Greyhounds and Labradors. At the stifle and tarsal joints, moment and power patterns were similar in shape, but amplitudes were larger for the Greyhounds. The MTP joint was a net absorber of energy, and this was greater in the Greyhounds. Greyhounds had a positive phase across the stifle, tarsal, and MTP joints at the end of stance for an active push-off, whereas for the Labrador Retrievers, the only positive phase was across the tarsus, and this was small, compared with values for the Greyhounds. CONCLUSIONS AND CLINICAL RELEVANCE: Gross differences in pelvic limb mechanics are evident between Greyhounds and Labrador Retrievers. Joint kinetics in specific dogs should be compared against breed-specific patterns. PMID- 16261831 TI - Effects of routine prophylactic vaccination or administration of aluminum adjuvant alone on allergen-specific serum IgE and IgG responses in allergic dogs. AB - OBJECTIVE: To determine the acute corn-specific serum IgE and IgG, total serum IgE, and clinical responses to s.c. administration of prophylactic vaccines and aluminum adjuvant in corn-allergic dogs. ANIMALS: 20 allergic and 8 nonallergic dogs. PROCEDURE: 17 corn-allergic dogs were vaccinated. Eight clinically normal dogs also were vaccinated as a control group. Serum corn-specific IgE, corn specific IgG, and total IgE concentrations were measured in each dog before vaccination and 1 and 3 weeks after vaccination by use of an ELISA. The corn allergic dogs also had serum immunoglobulin concentrations measured at 8 and 9 weeks after vaccination. Twenty allergic dogs received a s.c. injection of aluminum adjuvant, and serum immunoglobulin concentrations were measured in each dog 1, 2, 3, 4, and 8 weeks after injection. The allergic dogs were examined during the 8 weeks after aluminum administration for clinical signs of allergic disease. RESULTS: The allergic dogs had significant increases in serum corn specific IgE and IgG concentrations 1 and 3 weeks after vaccination but not 8 or 9 weeks after vaccination. Control dogs did not have a significant change in serum immunoglobulin concentrations after vaccination. After injection of aluminum adjuvant, the allergic dogs did not have a significant change in serum immunoglobulin concentrations or clinical signs. CONCLUSIONS AND CLINICAL RELEVANCE: Allergen-specific IgE and IgG concentrations increase after prophylactic vaccination in allergic dogs but not in clinically normal dogs. Prophylactic vaccination of dogs with food allergies may affect results of serologic allergen-specific immunoglobulin testing performed within 8 weeks after vaccination. PMID- 16261832 TI - Pharmacokinetics of azathioprine following single-dose intravenous and oral administration and effects of azathioprine following chronic oral administration in horses. AB - OBJECTIVE: To determine pharmacokinetics of azathioprine (AZA) and clinical, hematologic, and serologic effects of i.v. and oral administration of AZA in horses. ANIMALS: 6 horses. PROCEDURE: In study phase 1, a single dose of AZA was administered i.v. (1.5 mg/kg) or orally (3.0 mg/kg) to 6 horses, with at least 1 week between treatments. Blood samples were collected for AZA and 6 mercaptopurine (6-MP) analysis 1 hour before and at predetermined time points up to 4 hours after AZA administration. In study phase 2, AZA was administered orally (3 mg/kg) every 24 hours for 30 days and then every 48 hours for 30 days. Throughout study phase 2, blood samples were collected for CBC determination and serum biochemical analysis. RESULTS: Plasma concentrations of AZA and its metabolite, 6-MP decreased rapidly from plasma following i.v. administration of AZA, consistent with the short mean elimination half-life of 1.8 minutes. Oral bioavailability of AZA was low, ranging from 1% to 7%. No horses had abnormalities on CBC determination or serum biochemical analysis, other than 1 horse that was lymphopenic on day 5 and 26 of daily treatment. This horse developed facial alopecia from which 1 colony of a Trichophyton sp was cultured; alopecia resolved within 1 month after the study ended. CONCLUSIONS AND CLINICAL RELEVANCE: Overall, no adverse effects were observed with long-term oral administration of AZA to horses, although 1 horse did have possible evidence of immunosuppression with chronic treatment. Further investigation of the clinical efficacy of AZA in the treatment of autoimmune diseases in horses is warranted. PMID- 16261833 TI - Evaluation of cytokine production by equine alveolar macrophages exposed to lipopolysaccharide, Aspergillus fumigatus, and a suspension of hay dust. AB - OBJECTIVE: To evaluate cytokine production by equine alveolar macrophages after exposure to lipopolysaccharide (LPS), Aspergillus fumigatus, and hay dust, and determine the effect of clenbuterol on the cytokine response. ANIMALS: 6 horses. PROCEDURE: Alveolar macrophages were exposed to PBS solution (negative control), LPS, hyphae and conidia of Aspergillus fumigatus (AF), or a suspension of hay dust (HDS) and incubated for 24 hours at 37 degrees C. Concentrations of tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta were measured in the supernatant. The procedure was repeated with cells that were concurrently incubated with 0.5 microM clenbuterol. RESULTS: Exposure to HDS and AF significantly increased production of TNF-alpha by equine alveolar macrophages. The increase in TNF-alpha produced in response to HDS and AF was 5 and 7 times as great, respectively, as the increase measured in response to LPS. The concentration of IL-1beta in the supernatant was significantly increased after exposure of cells to AF. Clenbuterol was effective at inhibiting TNF-alpha production by cells exposed to LPS, HDS, or AF. CONCLUSIONS AND CLINICAL RELEVANCE: Increased production of TNF-alpha and IL-1 indicated that the pro inflammatory cytokines produced by alveolar macrophages in response to allergens may play a role in recurrent airway obstruction (RAO) in horses. Equine alveolar macrophages are not only a primary pulmonary defense mechanism but may also influence the pathogenesis of equine RAO. The beta2-adrenoceptor agonist clenbuterol, a drug that is commonly used for treatment of equine RAO, promotes immediate bronchodilation and may also contribute to downward modulation of the inflammatory response. PMID- 16261835 TI - Physiologic assessment of blood glucose homeostasis via combined intravenous glucose and insulin testing in horses. AB - OBJECTIVE: To characterize the physiologic response to i.v. bolus injection of glucose and insulin for development of a combined glucose-insulin test (CGIT) in horses. ANIMALS: 6 healthy mares and 1 mare each with pituitary adenoma and urolithiasis. PROCEDURE: Horses were given a CGIT (glucose, 150 mg/kg; insulin, 0.1 U/kg); results were compared with a singular i.v. glucose tolerance test (GTT; 150 mg/kg) and a singular i.v. insulin sensitivity test (IST; 0.1 U/kg). Healthy horses were also given a CGIT after receiving xylazine and undergoing stress. RESULTS: Physiologically, the CGIT resulted in a 2-phase curve with positive (hyperglycemic) and negative (hypoglycemic) portions; the positive phase came first (250% of baseline at 1 minute). The descending segment declined linearly to baseline by approximately 30 minutes and to a nadir at 58% of baseline by 75 minutes. After a 35-minute valley, a linear ascent to baseline began. Addition of insulin in the CGIT increased glucose utilization by approximately 4.5 times during the positive phase but not during the negative phase. The diseases' effects and experimental inhibition of insulin secretion with xylazine and stress were detectable by use of the 2 phases of the CGIT. Only a single positive phase resulted from the GTT and a single negative phase from the IST CONCLUSIONS AND CLINICAL RELEVANCE: The CGIT resulted in a consistent, well-defined glycemia profile, which can be disrupted experimentally or by a disease process. The CGIT has clinical potential because it provides integrated information and more information than either the singular GTT or IST. PMID- 16261834 TI - Cytokine response of bovine mammary gland epithelial cells to Escherichia coli, coliform culture filtrate, or lipopolysaccharide. AB - OBJECTIVE: To define the cytokine response of a cultured mammary gland epithelial cell line (ie, Mac-T) when incubated with Escherichia coli or its products. SAMPLE POPULATION: Mac-T cells and E coli from cows with mastitis. PROCEDURE: Mac T cells were incubated with E coli or its products. The cytokine response of Mac T cells to these treatments was quantified by measuring mRNA content of interleukin (IL)-1alpha, IL-1beta, IL-8, and tumor necrosis factor (TNF)-alpha by use of a quantitative reverse transcriptase-polymerase chain reaction assay. The amount of TNF-alpha secreted was also measured. RESULTS: Treatment with E coli or its products resulted in significant increases in IL-1alpha, IL-8, and TNF-alpha mRNA content in Mac-T cells. This increase was reversible when culture filtrate was incubated with polymyxin B. The amount of IL-1beta mRNA in Mac-T cells increased only slightly over baseline after treatment with E coli or its products, but this increase was not diminished by incubation of E coli filtrate with polymyxin B. CONCLUSIONS AND CLINICAL RELEVANCE: Incubation of Mac-T cells with E coli or its products resulted in increased amounts of IL1alpha, IL-8, and TNF-alpha mRNA. Inhibition of this response by incubation of culture filtrate with polymyxin B suggested that lipopolysaccharide was the main bacterial product that stimulated the cytokine response. The small increase in IL-1beta content in Mac-T cells incubated with E coli or its products suggested that this cytokine had a smaller role in the Mac-T cell response to E coli. PMID- 16261836 TI - Validation of pressure measurements and electromyographic results for the uterus of cattle during the early postpartum period. AB - OBJECTIVE: To evaluate methods for on-farm measurements of uterine contractility in postpartum dairy cows by comparing data simultaneously recorded by use of 2 intrauterine pressure (IUP) devices and quantified electromyographic (EMG) signals. ANIMALS: 5 cows during the first 48 hours after parturition. PROCEDURE: 2 EMG electrodes were implanted on the surface of the gravid uterine horn. Parturition was induced by injection of a prostaglandin F2alpha analogue at day 274 of gestation. An open-tip catheter and pressure microtransducer were transcervically inserted and affixed to a caruncle immediately after calving. Changes in IUP were recorded concurrent with EMG recordings during 2-hour periods at 2, 6, 12, 18, 24, 36, and 48 hours after parturition. Novel acquisition and analysis software programs were used with a digital data-filtering capability for evaluation of IUP and EMG signals. RESULTS: The method for intrauterine fixation of the 2 pressure measurement instruments was effective and allowed easy, externally guided removal of the devices 48 hours after parturition. There was a high correlation between the data obtained by the 2 pressure measuring systems. Good correlation was also found between pressure data obtained by the open-tip catheter system and EMG signals. Although the quantified IUP and EMG signals were highly comparable, synchronization was not always evident during visual inspection of these signals. CONCLUSIONS AND CLINICAL RELEVANCE: The open-tip IUP catheter system with a special fixation method is suitable for use in on-farm studies. It will enable investigators to record natural and pharmacologically influenced uterine contractility in early postpartum dairy cows. PMID- 16261837 TI - Assessment of a von Frey device for evaluation of the antinociceptive effects of morphine and its application in pharmacodynamic modeling of morphine in dogs. AB - OBJECTIVE: To assess the use of a von Frey device as a mechanical nociceptive stimulus for evaluation of the antinociceptive effects of morphine in dogs and its potential application in the pharmacodynamic modeling of morphine in that species. ANIMALS: 6 healthy Beagles. PROCEDURE: von Frey thresholds were measured in all dogs before and at intervals after they received no treatment (control dogs) and i.v. administration of morphine sulfate (1 mg/kg; treated dogs) in a crossover study. The von Frey device consisted of a rigid tip (0.5 mm in diameter) and an electronic load cell; the operator was unaware of recorded measurements. RESULTS: Application of the von Frey device was simple and well tolerated by all dogs and caused no apparent tissue damage. No significant changes in thresholds were detected in the control dogs at 8 hourly measurements, indicating a lack of acquired tolerance, learned aversion, or local hyperalgesia. When assessed as a group, treated dogs had significantly high thresholds for 4 hours following morphine administration, compared with baseline values; individually, thresholds decreased to baseline values within (mean +/- SE) 2.8 +/ 0.6 hours. The maximal effect (change from baseline values) was 213 +/- 43%, and the plasma morphine concentration to achieve 50% maximal effect was 13.92 +/- 2.39 ng/mL. CONCLUSIONS AND CLINICAL RELEVANCE: Data suggest that, in dogs, evaluation of the antinociceptive effect and pharmacodynamic modeling of a dose of morphine sulfate (1 mg/kg, i.v.) can be successfully achieved by use of a von Frey device. PMID- 16261838 TI - Functional adaptation through changes in regional biochemical characteristics during maturation of equine superficial digital flexor tendons. AB - OBJECTIVE: To quantify and compare biochemical characteristics of the extracellular matrix (ECM) of specimens harvested from tensional and compressive regions of the superficial digital flexor tendon (SDFT) of horses in age classes that include neonates to mature horses. SAMPLE POPULATION: Tendon specimens were collected on postmortem examination from 40 juvenile horses (0, 5, 12, and 36 months old) without macroscopically visible signs of tendonitis. PROCEDURE: Central core specimens of the SDFT were obtained with a 4-mm-diameter biopsy punch from 2 loaded sites, the central part of the mid-metacarpal region and the central part of the mid-sesamoid region. Biochemical characteristics of the collagenous ECM content (ie, collagen, hydroxylysylpyridinoline crosslink, and pentosidine crosslink concentrations and percentage of degraded collagen) and noncollagenous ECM content (percentage of water and glycosaminoglycans, DNA, and hyaluronic acid concentrations) were measured. RESULTS: The biochemical composition of equine SDFT was not homogeneous at birth with respect to DNA, glycosaminoglycans, and pentosidine concentrations. For most biochemical variables, the amounts present at birth were dissimilar to those found in mature horses. Fast and substantial changes in all components of the matrix occurred in the period of growth and development after birth. CONCLUSIONS AND CLINICAL RELEVANCE: Unlike cartilage, tendon tissue is not biochemically blank (ie, homogeneous) at birth. However, a process of functional adaptation occurs during maturation that changes the composition of equine SDFT from birth to maturity. Understanding of the maturation process of the juvenile equine SDFT may be useful in developing exercise programs that minimize tendon injuries later in life that result from overuse. PMID- 16261839 TI - Evaluation of well-being, productivity, and longevity of pregnant sows housed in groups in pens with an electronic sow feeder or separately in gestation stalls. AB - OBJECTIVE: To compare well-being, performance, and longevity of gestating sows housed in stalls or in pens with an electronic sow feeder (ESF). ANIMALS: 382 pregnant sows of parities 1 through 6. PROCEDURE: Sows were housed in separate stalls (n = 176) or group pens (206) with an ESF. Well-being of sows was assessed at various time points in terms of injuries, salivary cortisol concentration, and behavior in a novel arena or to a novel object. Farrowing performance and longevity of sows were also assessed. RESULTS: Total injury scores (TIS) of sows in pens were significantly higher at initial introduction and mixing. In stall housed sows, TIS was significantly higher during late gestation. The TIS and cortisol concentration were significantly lower in stall-housed sows, compared with values for sows in pens. As parity increased, the likelihood of higher median TIS decreased significantly in pen-housed sows and increased significantly in stall-housed sows. The TIS of sows in pens was negatively correlated with body weight and backfat thickness, whereas these correlations were positive in stall housed sows. Farrowing performance and results for novel arena or objects did not differ. Proportion of sows removed was significantly higher for pens than for stalls; lameness was the major reason for removal for both systems. CONCLUSIONS AND CLINICAL RELEVANCE: Stalls impose space restrictions for larger sows, resulting in injuries during late gestation. Interventions are needed to minimize aggression during initial introduction and mixing and at the ESF in pens to reduce severe injuries or lameness of gestating sows. PMID- 16261840 TI - Evaluation of a lithium dilution cardiac output technique as a method for measurement of cardiac output in anesthetized cats. AB - OBJECTIVE: To evaluate the use of a lithium dilution cardiac output (LiDCO) technique for measurement of CO and determine the agreement between LiDCO and thermodilution CO (TDCO) values in anesthetized cats. ANIMALS: 6 mature cats. PROCEDURE: Cardiac output in isoflurane-anesthetized cats was measured via each technique. To induce different rates of CO in each cat, anesthesia was maintained at > 1.5X end-tidal minimum alveolar concentration (MAC) of isoflurane and at 1.3X end-tidal isoflurane MAC with or without administration of dobutamine (1 to 3 microg/kg/min, i.v.). At least 2 comparisons between LiDCO and TDCO values were made at each CO rate. The TDCO indicator was 1.5 mL of 5% dextrose at room temperature; with the LiDCO technique, each cat received 0.005 mmol of lithium/kg (concentration, 0.015 mmol/mL). Serum lithium concentrations were measured prior to the first and following the last CO determination. RESULTS: 35 of 47 recorded comparisons were analyzed; via linear regression analysis (LiDCO vs TDCO values), the coefficient of determination was 0.91. The mean bias (TDCO-LiDCO) was -4 mL/kg/min (limits of agreement, -35.8 to + 27.2 mL/kg/min). The concordance coefficient was 0.94. After the last CO determination, serum lithium concentration was < 0.1 mmol/L in each cat. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated a strong relationship and good agreement between LiDCO and TDCO values; the LiDCO method appears to be a practical, relatively noninvasive method for measurement of CO in anesthetized cats. PMID- 16261841 TI - Life expectancy in a birth cohort of Boxers followed up from weaning to 10 years of age. AB - OBJECTIVE: To determine mortality rate over time, risk factors for death, and heritability of life expectancy in Boxers. ANIMALS: 1,733 purebred Boxers born in The Netherlands between January 1994 and March 1995. PROCEDURE: Dogs were followed up from weaning (ie, 49 days of age) to 10 years of age through use of a written questionnaire sent to owners every 6 months. Mortality rate over time, risk factors potentially associated with death, and heritability of life expectancy were examined by use of a proportional hazards model based on the Weibull distribution. RESULTS: stimated mortality rate during the 10-year study period for this birth cohort of Boxers was 45%. The probability of surviving to 5 years of age was 88%; the probability of surviving to 10 years of age was 55%. Estimated effective heritability of life expectancy was 0.076, meaning that in this population, an estimated 76% of the observed variation in life expectancy could be attributed to genetic differences among dogs that were passed from parents to their offspring. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that cumulative incidence of death from weaning to 10 years of age among this birth cohort of Boxers was 45%. The estimated heritability of life expectancy suggested that life expectancy can be improved by use of selective breeding. PMID- 16261842 TI - In vitro evaluation of canine and feline calcium oxalate urolith fragility via shock wave lithotripsy. AB - OBJECTIVE: To test the hypothesis that feline calcium oxalate uroliths are intrinsically more resistant to comminution via shock wave lithotripsy (SWL) than canine calcium oxalate uroliths through comparison of the fragility of canine and feline uroliths in a quantitative in vitro test system. SAMPLE POPULATION: Calcium oxalate uroliths (previously obtained from dogs and cats) were matched by size and mineral composition to create 7 pairs of uroliths (1 canine and 1 feline urolith/pair). PROCEDURE: Uroliths were treated in vitro with 100 shock waves (20 kV; 1 Hz) by use of an electrohydraulic lithotripter. Urolith fragmentation was quantitatively assessed via determination of the percentage increase in projected area (calculated from the digital image area of each urolith before and after SWL). RESULTS: After SWL, canine uroliths (n = 7) fragmented to produce a mean +/ SD increase in image area of 238 +/- 104%, whereas feline uroliths (7) underwent significantly less fragmentation (mean image area increase of 78 +/- 97%). The post-SWL increase in fragment image area in 4 of 7 feline uroliths was < 50%, whereas it was > 150% in 6 of 7 canine uroliths. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate that feline calcium oxalate uroliths are less susceptible to fragmentation via SWL than canine calcium oxalate uroliths. In some cats, SWL may not be efficacious for fragmentation of calcium oxalate nephroliths or ureteroliths because the high numbers of shock waves required to adequately fragment the uroliths may cause renal injury. PMID- 16261843 TI - Gene therapeutic exploration: retrovirus-mediated soluble vascular endothelial growth factor receptor-2 (sFLK-1) inhibits the tumorigenicity of S180, MCF-7, and B16 cells in vivo. AB - Inhibition of tumor angiogenesis is an anticancer strategy in which neovasculature is targeted because tumor progression relies on neovascularization. The soluble, truncated form of vascular endothelial growth factor receptor-2 (VEGFR-2), sFLK-1, is a well-known inhibitor of endothelial cells. This kind of soluble receptor retains its high-affinity binding to VEGF, but cannot work with the receptor tyrosine transphosphorylation and activation of downstream signal transduction to induce endothelial proliferation due to the lack of the tyrosine kinase domain. Therefore, we tried to use this sFLK-1 as an inhibitor for malignant tumor gene therapy. In this study we transferred a soluble VEGFR-2 (sFLK-1) from embryo mouse liver by RT-PCR to PA317 cells through retroviral vector (pLXSN) and obtained stable expression. NIH3T3 cells were used for measuring the virus titer. The virus titer in this experiment was 2 x 10(7) CFU/ml. After 7 days of preparing subcutaneous tumor models bearing S180, MCF-7, and B16 cells in mice, respectively, 2 x 10(7) PFU of recombinant retroviruses were injected locally into the tumors the treatment groups. After treatment, the tumor size and weight were significantly smaller than that of control (p < 0.05). After autopsy, the metastasic focus numbers in the treatment groups were also less than control groups. We also measured VEGFR-2 expression in tumor tissues by Western blot to check if sFLK-1 had been integrated into the cells of tumor tissues. Expression in the treatment groups was significantly greater than that of control groups (p < 0.001). Microvessel density (MVD) and proliferative cell nuclear antigen (PCNA) were investigated to determine whether the Re-sFLK-1 fragment had the ability to inhibit tumor angiogenesis and proliferation in mice bearing S180 and MCF-7 cells. The results showed that MVD and PCNA in th e treatment groups werelower than in control groups. There were significant difference between treatment groups and control groups (p < 0.0001). The results indicated that retroviral-mediated sFLK-1 gene therapy in animal tumor models has significant therapeutic effect. PMID- 16261844 TI - Bcl-2 as prognostic factor in head and neck squamous cell carcinoma. AB - A series of 66 cases of oral squamous cell carcinoma (OSCC) was retrospectively analyzed by immunohisto-chemistry for bcl-2 expression to verify its predictive value for clinical outcome in patients with OSCC. After grouping for bcl-2 expression, OSCCs were statistically analyzed for the variables age, gender, histological grading (G), TNM, staging, recurrence, and overall survival rate. Univariate and multivariate (Cox regression) analyses were performed. Thirty-six OSCC (54.5%) showed expression for bcl-2, whereas 30 (44.5%) were negative. No statistical association was found between bcl-2 expression and any variables considered at baseline. Overall disease-specific survival rate at 72 months was 51%, independently from the extent of the tumor. In terms of prognostic significance, the bcl-2-positive group showed more than 60% survival at 72 months whereas the bcl-2-negative group showed none. An independent association of bcl-2 expression was found with an improved overall survival rate (p = 0.048), although grading and staging were established to be the best baseline markers of prognosis. On the basis of these results, it is possible to suggest bcl-2 as an early marker of prognosis: lack of bcl-2 expression could constitute a hallmark of aggressive biological behavior in OSCC. PMID- 16261845 TI - Hypomethylation of the XIST gene promoter in prostate cancer. AB - In a process denoted "global hypomethylation" repetitive DNA sequences like LINE 1 retrotransposons become hypomethylated in human cancers, including a subset of prostate carcinomas. It is less well known to what extent single-copy sequences are affected by this phenomenon. Therefore, we have analyzed methylation and expression of the XIST gene by bisulfite sequencing and real-time RT-PCR. The promoter of this single-copy gene is strongly methylated in normal male cells, including leukocytes and normal prostate. In prostate cancer tissues and particularly in cell lines, partial hypomethylation was observed paralleling that of LINE-1 sequences. Weak XIST expression was found in normal prostate tissues, but none in leukocytes. Only slight increases in expression of this gene were found in cancer tissues and cell lines. Our data suggest that hypomethylation in prostate cancer is indeed "global," affecting repeat and unique sequences in parallel. Detection of partially hypomethylated XIST alleles in prostate cancer tissues might be useful for the identification of cases with pronounced hypomethylation, which tend to be more aggressive. PMID- 16261847 TI - The use of complementary and alternative therapies by patients with cancer. AB - The aim of this research was to assess the prevalence and predictors of complementary and alternative therapy (CAT) use among cancer patients in Australia. A total of 1492 cancer patients attending nine major public cancer treatment centers in New South Wales, Australia, were asked to complete the Supportive Care Needs Survey. Of the 1354 consenting patients, 888 (65%) returned a completed survey. This article reports the secondary analyses of the survey data, specifically focusing on CAT use. For all cancers, 17.1% of patients were using at least one CAT. The two main demographic characteristics of CAT users were gender and age, where females were more likely to use CAT than males and that CAT use declined as age increased. Time since diagnosis was identified as the only significant clinical predictor of CAT use, where CAT use increased with time until 5 years since diagnosis. Our research shows that herbal treatments and naturopathy are the most popular CAT used by cancer patients (constituting over 30% of all CAT use recorded). The use of CAT among cancer patients is a significant issue in cancer care, especially considering the potential interactions between CAT and conventional medicines. Given that many cancer patients may not be aware of potential risks associated with these interactions it is important that oncologists and others involved in cancer patient care are informed about CAT and its use amongst their patients. PMID- 16261846 TI - A bisanthracycline (WP631) represses uPAR gene expression and cell migration of RKO colon cancer cells by interfering with transcription factor binding to a chromatin-accessible -148/-124 promoter region. AB - The urokinase receptor (uPAR), transcriptionally activated in several cancers, contributes to tumor progression by promoting cell migration and proteolysis, and repressing expression of this gene could be of therapeutic utility. Indeed, targeting regulatory element(s) in the promoter may represent an efficient means for reducing expression because only two alleles have to be neutralized. We previously identified the -148/-124 promoter region, bound with Sp1 and Sp3, as regulatory for uPAR expression in vitro. The purpose of this study was twofold: to determine (a) the accessibility of this region in its natural chromatin setting and (b) the efficacy of WP631, a bisintercalator favoring GC-rich DNA sequences, in repressing endogenous uPAR expression in RKO colon cancer cells. In these cells, DNaseI hypersensitivity, genomic footprinting, and chromatin immunoprecipitation experiments revealed that the -148/-124 uPAR promoter region was accessible in chromatin and bound with Sp1, thus validating it as a therapeutic target. WP631 treatment competed for transcription factor binding to this regulatory region and reduced uPAR mRNA/protein. However, a chemically related compound (WP629), with low DNA binding affinity, failed to diminish uPAR protein amount. GAPDH mRNA level was only modestly affected by WP631, arguing against the possibility that this bisanthracycline universally represses expression of GC-rich promoter-driven genes. Further, uPAR function, as assessed by migration of cells across a vitronectin-coated filter, was attenuated with WP631. Thus, we have shown that the chromatinized -148/-124 regulatory region of the uPAR promoter is accessible to small molecules and that WP631, which disrupts the interaction of DNA binding proteins with this region, diminishes uPAR expression and function. PMID- 16261848 TI - Keyed vaporizer refilling not infallible. PMID- 16261849 TI - Area health education centers and health education training centers: a well-kept secret. PMID- 16261850 TI - The certified registered nurse anesthetist: occupational responsibilities, perceived stressors, coping strategies, and work relationships. AB - A qualitative inquiry was launched to explore occupational stress among Certified Registered Nurse Anesthetists (CRNAs). The purpose of this study was to examine how job-related stress manifests itself among CRNAs regarding their ability to relate to their peers. Twenty CRNAs and 15 of their coworkers from North Carolina and Tennessee participated in the study. To help confirm emerging findings, data triangulation (ie, semistructured interviews, clinical observations, and artifact data) was used to answer 4 research questions. Perceived occupational-related stressors identified by the CRNAs pertained to patient care, anesthesia work in general, job relationships, inadequate surgical preparation, the operating room environment, and physical stressors. Staying focused on patient care, the use of humor, verbalization and internalization of concerns, and adopting personal hobbies were identified as coping mechanisms to combat work-related stress. Moreover, 6 major themes surfaced after analyzing the data using the constant comparative method. The findings underscore that the shortage of registered nurses and anesthetists needs to be addressed to more effectively tackle the participants' perceived stressors. Employers can adopt concrete measures in assisting CRNAs with handling occupational stress, such as offering mandatory in servicing and adequate time to attend in-servicing. PMID- 16261851 TI - Dexmedetomidine as a sole sedating agent with local anesthesia in a high-risk patient for axillofemoral bypass graft: a case report. AB - The alpha2-agonist dexmedetomidine is indicated for sedation of patients receiving mechanical ventilation in the intensive care unit. It has additional off-label uses for coadministration with local, regional, and general anesthesia. This report describes the use of dexmedetomidine as a sole sedating agent in conjunction with local anesthesia for major vascular surgery. A PubMed literature search produced no previous report of the use of dexmedetomidine as a sole sedating agent used in conjunction with local anesthesia. The anxiolytic, hypnotic-sedative, anesthetic-sparing, and analgesic actions of the drug along with the lack of significant respiratory depressant effects are described. The patient required no airway management with the exception of supplemental mask oxygen. He tolerated the procedure well and was discharged without sequelae on the third postoperative day. Dexmedetomidine should be used judiciously, and understanding the potential adverse effects and how to treat them is of paramount importance. However, with vigilant intraoperative monitoring of blood pressure, heart rate, and level of consciousness, it can be administered safely, thus lessening the anesthetic requirements and possibly improving the surgical outcome of the high-risk patient. This report describes the indications, dosing, off label uses, pharmacodynamics, pharmacokinetics, and common adverse effects of dexmedetomidine. PMID- 16261852 TI - SNOMED CT: electronic health record enhances anesthesia patient safety. AB - This article discusses the importance of electronic health records (EHRs) in anesthesiology, emphasizing the critical role of standardized clinical terminology in the EHR's structure. SNOMED Clinical Terms (SNOMED CT), developed by SNOMED International, a division of the College of American Pathologists, in collaboration with the United Kingdom's National Health Service, offers a controlled healthcare terminology with comprehensive coverage of diseases, clinical findings, etiologies, therapies, procedures, and outcomes. Recommended as the core general terminology for electronic patient medical record information in the United States, it offers flexibility in expressing clinical concepts, enabling clinicians to say things in multiple ways and still be understood. SNOMED CT's comprehensive, scientifically validated clinical terminology enables a consistent way of capturing, sharing, and aggregating health data across specialties and sites of care. Its benefits range from facilitating system interoperability to allowing greater shared access to patient health information where and when it is needed. The article concludes by informing readers of access to English and Spanish language editions of SNOMED CT's core content, which recently was licensed through the National Library of Medicine, a part of the National Institutes of Health within Department of Health and Human Services. PMID- 16261853 TI - A national survey of certified registered nurse anesthetists' knowledge, beliefs, and assessment of herbal supplements in the anesthesia setting. AB - This study explored Certified Registered Nurse Anesthetists' (CRNAs') knowledge about 8 common herbal supplements that potentially cause perioperative complications, beliefs regarding herbal supplement-anesthesia interactions, and preoperative assessment practices. A multiple-choice descriptive survey was mailed to a random sample of CRNAs from the American Association of Nurse Anesthetists (AANA). Although response was low (19%, N = 191), respondent demographics corresponded to AANA 2002 CRNA membership. The mean knowledge score of adverse interactions was 21%; 17% of CRNAs indicated confidence in their familiarity with herbal supplement-anesthesia interactions. Of the participants, 23% correctly identified the ASA recommendation to discontinue herbal supplements 2 weeks before surgery. CRNAs agreed that herbal supplement use should be assessed preoperatively (92%), they are medically active (88%), and they can have an impact on surgical outcomes (87%). Nearly 4 in 10 respondents used herbal supplements themselves, but only 5% recommend supplement use to patients. Most CRNAs (93%) wanted more educational opportunities on anesthesia and herbal supplements. The low knowledge scores and lack of confidence in familiarity with herbal supplement-anesthesia interactions highlight the need for further CRNA education. Nurse anesthesia educational curricula and continuing education programs should be reviewed and updated to ensure adequate instruction on herbal supplements. PMID- 16261854 TI - Acupressure and postoperative nausea and vomiting. AB - Despite great strides during the preceding 3 decades, the ability to consistently eliminate postoperative nausea and vomiting (PONV) continues to elude anesthesia practitioners. The occurrence of PONV related to anesthesia and surgery prolongs hospital stays and increases healthcare costs. Protracted recovery times place constraints on patients, healthcare systems, and healthcare financiers. Many pharmacological antiemetics have been developed and are in use in the attempt to alleviate PONV. Side effects and cost profiles of many of these interventions, however, reinforce the broadly held belief that there remains opportunity for improvement. Because the Western culture almost exclusively favors evidence-based scientific practice and interventions, the search continues for an ideal, cost effective, safe, and efficacious pharmacological agent to prevent PONV. Eastern culture, on the other hand, relies heavily on naturopathic remedies whose successful use has spanned thousands of years. Increasing attention has been given to the potential benefits of nonpharmacological intervention for the prevention of PONV in association with anesthesia care. Therefore, the purpose of this AANA Journal course will be to focus attention on what is known and what is unknown in the literature regarding use of the nonallopathic remedy of acupressure as a nonpharmacological alternative to commonly utilized antiemetic prophylaxis. PMID- 16261855 TI - Differential inactivation of glucose- and glutamate dependent acid resistance of Escherichia coli TMW 2.497 by high-pressure treatments. AB - The inactivation by 200-400 MPa and post-pressure survival at acid conditions of E. coli TMW 2.497 was characterized by the measurement of intracellular pH (pHin), viable cell counts, glutamate (Glu) and arginine (Arg) consumption, and the influence of mild adaptation to mild acid stress prior to pressure treatment. Glutamate and arginine did not affect viable cell counts or the pHin during pressure application but improved the ability to maintain a high pHin after pressure treatment. In pH 4.0 buffer without arg and glu, a 3 log reduction of cell counts occurred after 24 h of incubation, whereas little or no loss of viability was observed after 24 h incubation in the presence of glu and arg. During post-pressure incubation at pH 4.0, 10 mM glutamate were metabolized but only 2 mM arginine were used, indicating that glutamate rather than arginine was responsible for the protective effect on pHin and survival. In conclusion, the pressure induced, irreversible loss of the transmembrane deltapH correlates to cell death and glu stabilizes the pHin of E. coli during post-pressure incubation. PMID- 16261856 TI - Stappia alba sp. nov., isolated from Mediterranean oysters. AB - Three bacterial strains isolated from oysters recovered at the Spanish Mediterranean coast have been phenotypically and genetically characterized. The results of the phylogenetic analysis based on almost complete 16S rDNA sequences clustered all three strains together with 99.9% average sequence similarity and situated them in the neighbourhood of the genera Stappia, Roseibium and Pannonibacter, Stappia aggregata being their closest neighbour with sequence similarities between 98.8% and 98.9%. DNA-DNA hybridization experiments using DNA of strains 5OM6T and S. aggregata CECT 4269T as reference DNAs confirmed the independent status at species level of the oyster isolates. Phenotypically, they can be distinguished from the closest relatives by the ionic requirements, growth temperatures and use of carbon compounds. We propose these oyster strains constitute a new species of Stappia, for which the name Stappia alba sp. nov. has been chosen, and strain 5OM6T (= CECT 5095T = CIP 108402T) as its type strain. PMID- 16261857 TI - Citreicella thiooxidans gen. nov., sp. nov., a novel lithoheterotrophic sulfur oxidizing bacterium from the Black Sea. AB - Four strains of obligately heterotrophic bacteria isolated from the oxygen sulfide interface of the Black Sea are characterized. The bacteria are aerobic, Gram-negative, with lemon-like, nonmotile cells. Bacteriochlorophyll a is not detected. They are mesophilic and neutrophilic with a temperature range of 8-35 degrees C (optimum 25) and pH range of 6.5-8.5 (optimum 7.8). Their growth is NaCl dependent within a range of 5 and 60 (optimum 20) g l(-1). They are able to oxidize thiosulfate, sulfide and elemental sulfur to sulfate and to use metabolic energy from these reactions (lithoheterotrophy). According to the level of DNA reassociation of more than 40%, all isolates represent a single generic group. The G+C content of the DNA was in the range of 67.5-69.2mol%. According to phylogenetic analysis, the new isolates form a separate branch in the alpha-3 subdivision of the Proteobacteria together with two undescribed marine bacterial strains. On the basis of phenotypical and genomic properties, the new isolates are described as a new genus and species Citreicella thiooxidans gen. nov., sp. nov. The type strain is CHLG 1T ( = DSM 10146, UNIQEM U 228). PMID- 16261858 TI - Lactobacillus harbinensis sp. nov., consisted of strains isolated from traditional fermented vegetables 'Suan cai' in Harbin, Northeastern China and Lactobacillus perolens DSM 12745. AB - Taxonomical analysis of two genetically distinguished Lactobacillus strains isolated from traditional Chinese fermented vegetables 'Suan cai' was performed. They formed L-lactate from glucose, were facultatively heterofermentative, and had a DNA G+C content of 53-54mol%. They fermented D- and L-arabinose. They produced lactate, ethanol and acetate from gluconate at a molar ratio of 1.1:0.4:0.7. Phylogenetic analysis of 16S rDNA revealed that the two strains were closely related to L. perolens. DNA-DNA hybridization analysis revealed that the two strains were different from L. perolens type strain DSM 12744 and formed a separate cluster with L. perolens DSM 12745. G + C molar content of DNA of the former is 51%, whereas those of the latter strains were in the range of 53-54%. Based on the results, we propose that the new species be named L. harbinensis sp. nov. and that L. perolens DSM 12745 be reclassified as L. harbinensis DSM 12745. The type strain of L. harbinensis DSM 16991T (= AHU 1762T = SBT 10908T). PMID- 16261859 TI - Rhodococcus phenolicus sp. nov., a novel bioprocessor isolated actinomycete with the ability to degrade chlorobenzene, dichlorobenzene and phenol as sole carbon sources. AB - The aerobic degradation of phenol, chlorobenzene and dichlorobenzene as a sole carbon source has been observed in bacterial Gram-positive strain G2PT isolated from a wastewater bioprocessor. Cells display branching mycelia fragmenting into rod and coccoid elements when grown on TSA. Aerial hyphae formation occurs when grown on phenol and chlorinated aromatics as the sole carbon source. Growth was observed at up to 0.75% phenol as a sole carbon source, indicating a strong tolerance for the compound. The 16S rRNA gene sequence shares the greatest similarity with members of the Rhodococcus genus, with the closest shared nucleotide identity of 98% with the aromatic toxin degrading bacteria Rhodococcus zopfii DSM 44108T. Neighbor-joining and parsimony analysis of Corynebacterineae 16S rRNA gene sequences consistently places strain G2PT in a clade shared with R. zopfii within the Rhodococcus rhodochrous subclade. Based on a unique polyphasic profile involving phenotypic, ribosomal DNA sequence analysis, DNA-DNA hybridization, mol% DNA G+C content and fatty acid composition, G2PT is proposed to represent a previously uncharacterized, novel species in the genus Rhodococcus. The name Rhodococcus phenolicus is proposed for the isolate with the type strain G2PT (= DSM 44812) (= NRRL B-24323) [corrected] PMID- 16261861 TI - Analysis of yeast isolates related to Metschnikowia pulcherrima using the partial sequences of the large subunit rDNA and the actin gene; description of Metschnikowia andauensis sp. nov. AB - Thirty-two yeast isolates were cultured from guts or excrements of three different pests of corn or from the stem of healthy corn. The strains were analyzed using MSP-PCR (micro/minisatellite-primed polymerase chain reaction), sequences of the D1/D2 region of the large subunit rDNA and a 979 bp long part of the actin gene (act-1). They seem to belong to three groups that are all sister groups of Metschnikowia pulcherrima, M. fructicola and M. chrysoperlae. A new species, Metschnikowia andauensis (HA 1657T) is described. In contrast to M. pulcherrima and M. fructicola, M. andauensis is well separated in the act-1 phylogenetic tree too. PMID- 16261860 TI - Molecular systematics of rhizobia based on maximum likelihood and Bayesian phylogenies inferred from rrs, atpD, recA and nifH sequences, and their use in the classification of Sesbania microsymbionts from Venezuelan wetlands. AB - A well-resolved rhizobial species phylogeny with 51 haplotypes was inferred from a combined atpD + recA data set using Bayesian inference with best-fit, gene specific substitution models. Relatively dense taxon sampling for the genera Rhizobium and Mesorhizobium was achieved by generating atpD and recA sequences for six type and 24 reference strains not previously available in GenBank. This phylogeny was used to classify nine nodule isolates from Sesbania exasperata, S. punicea and S. sericea plants native to seasonally flooded areas of Venezuela, and compared with a PCR-RFLP analysis of rrs plus rrl genes and large maximum likelihood rrs and nifH phylogenies. We show that rrs phylogenies are particularly sensitive to strain choice due to the high levels of sequence mosaicism found at this locus. All analyses consistently identified the Sesbania isolates as Mesorhizobium plurifarium or Rhizobium huautlense. Host range experiments on ten legume species coupled with plasmid profiling uncovered potential novel biovarieties of both species. This study demonstrates the wide geographic and environmental distribution of M. plurifarium, that R. galegae and R. huautlense are sister lineages, and the synonymy of R. gallicum, R. mongolense and R. yanglingense. Complex and diverse phylogeographic, inheritance and host association patterns were found for the symbiotic nifH locus. The results and the analytical approaches used herein are discussed in the context of rhizobial taxonomy and molecular systematics. PMID- 16261863 TI - Quick identification of acetic acid bacteria based on nucleotide sequences of the 16S-23S rDNA internal transcribed spacer region and of the PQQ-dependent alcohol dehydrogenase gene. AB - Acetic acid bacteria (AAB) are well known for oxidizing different ethanol containing substrates into various types of vinegar. They are also used for production of some biotechnologically important products, such as sorbose and gluconic acids. However, their presence is not always appreciated since certain species also spoil wine, juice, beer and fruits. To be able to follow AAB in all these processes, the species involved must be identified accurately and quickly. Because of inaccuracy and very time-consuming phenotypic analysis of AAB, the application of molecular methods is necessary. Since the pairwise comparison among the 16S rRNA gene sequences of AAB shows very high similarity (up to 99.9%) other DNA-targets should be used. Our previous studies showed that the restriction analysis of 16S-23S rDNA internal transcribed spacer region is a suitable approach for quick affiliation of an acetic acid bacterium to a distinct group of restriction types and also for quick identification of a potentially novel species of acetic acid bacterium (Trcek & Teuber 2002; Trcek 2002). However, with the exception of two conserved genes, encoding tRNAIle and tRNAAla, the sequences of 16S-23S rDNA are highly divergent among AAB species. For this reason we analyzed in this study a gene encoding PQQ-dependent ADH as a possible DNA-target. First we confirmed the expression of subunit I of PQQ-dependent ADH (AdhA) also in Asaia, the only genus of AAB which exhibits little or no ADH activity. Further we analyzed the partial sequences of adhA among some representative species of the genera Acetobacter, Gluconobacter and Gluconacetobacter. The conserved and variable regions in these sequences made possible the construction of A. acetispecific oligonucleotide the specificity of which was confirmed in PCR-reaction using 45 well-defined strains of AAB as DNA templates. The primer was also successfully used in direct identification of A. aceti from home made cider vinegar as well as for revealing the misclassification of strain IFO 3283 into the species A. aceti. PMID- 16261862 TI - Enumeration and characterization of culturable arsenate resistant bacteria in a large estuary. AB - Arsenic is a toxic element that exists in two major inorganic forms, arsenate and arsenite. A number of bacteria have been shown to resist arsenic exposure, and even more bacteria appear to possess the genes for arsenic resistance. In this study, the numbers of culturable arsenate-resistant bacteria present in water at three coastal sites in the Lake Pontchartrain estuary, Louisiana, was determined. Despite insignificant (less than 1.33 microM) levels of arsenic in this system, 20-50% of the viable count of bacteria showed appreciable arsenate resistance, suggesting that arsenic-resistant bacteria are common and widespread. A diverse array of arsenate-resistant isolates was obtained, with 16S rRNA sequence analysis indicating 37 different bacterial strains, representing six major bacterial groups. Many of these isolates were affiliated with groups of bacteria that have been poorly characterized in terms of arsenic resistance, such as the Betaproteobacteria or Flavobacteria. Some isolates were capable of tolerating very high (> 100 mM) levels of arsenate, although arsenite resistance was generally much lower. The results suggest that arsenic-resistant bacteria are common, even in environments with insignificant arsenic contamination, and that many different groups of aquatic bacteria show appreciable arsenic resistance. PMID- 16261864 TI - Halothiobacillus neapolitanus strain OSWA isolated from "The Old Sulphur Well" at Harrogate (Yorkshire, England). AB - The "Old Sulphur Well" has a subterranean input of water containing 5.5mM total sulfide, which would be inhibitory to the growth of most bacteria. The obligately chemolithoautotrophic Halothiobacillus neapolitanus is a sulfur bacterium known to tolerate and metabolize high sulfide concentrations, and we report the isolation of H. neapolitanus strain OSWA from this source. Strain OSWA grows well on thiosulfate and tetrathionate as energy sources, and tolerates at least 5mM sulfide. Its specific growth rates and yields in batch culture were 0.22h(-1) and 5.3 gmol(-1) (thiosulfate), and 0.23 h(-1) and 9.5 gmol(-1) (tetrathionate). Its 16S rRNA gene sequence shows >99% identity to reference sequences of H. neapolitanus, and it shares morphological and physiological characteristics typical of the species. It is one of a very small number of strains of H. neapolitanus described to date, and the first to be isolated from an ancient sulfide-rich natural spa. PMID- 16261865 TI - Mycosporines in carotenogenic yeasts. AB - The ability to produce mycosporines (MYCs) in 157 pigmented yeast strains (eight genera, 25 species) isolated from natural environments of Patagonia (Argentina) was assessed. The strains belong to four taxonomic groups: the Sporidiobolales and Erythrobasidium clade of the class Urediniomycetes, and Cystofilobasidiales and Tremellales of the class Hymenomycetes. Induction of MYCs did not occur in all yeast strains tested and appeared to be an exclusive trait of members of the Erythrobasidium clade and Tremellales. This is the first report on the production of MYCs by pigmented species from the latter group, as well as the first extensive screening of mycosporinogenic yeasts. The consistent occurrence of MYCs in some specific phylogenetic groups suggests this trait bears evolutionary significance and that the presence/absence of MYCs may have potential applications in yeast systematics. PMID- 16261866 TI - Biodiversity of Saccharomyces cerevisiae isolated from a survey of pito production sites in various parts of Ghana. AB - Biodiversity among Saccharomyces cerevisiae predominating the spontaneous fermentation of Dagarti pito in Ghana was assessed. Two hundred and forty-nine isolates obtained from samples of dried yeast taken from commercial pito production sites in eight geographical regions of Ghana were characterized phenotypically by colony and cell morphology as well as carbohydrate assimilation profiling. Yeast populations ranged between 10(6) and 10(8) cfug(-1). Ninety-nine percent of the isolates (247) investigated showed macro-and micro morphological characteristics typical of S. cerevisiae. Of these, 72% (179) had assimilation profiles similar to S. cerevisiae while 28% (68) had assimilation profiles atypical of S. cerevisiae or any other member of the Saccharomyces sensu stricto complex. Amplification of the region spanning the two intergenic transcribed spacers (ITS) and the 5.8S ribosomal gene (ITS1-5.8S rDNA-ITS2), followed by restriction analysis, as well as determination of chromosome length polymorphism by pulsed field gel electrophoresis (PFGE) of 25 representative isolates strongly indicated that all belonged to S. cerevisiae, notwithstanding the phenotypic differences. Sequencing of the mitochondrial cytochrome-c oxidase II gene (COX 2) and the actin-encoding gene (ACT1) of four isolates, confirmed their close relatedness to S. cerevisiae, particularly to the type strain CBS1171 (98.7%), as well as other members of the Saccharomyces sensu stricto complex. Twenty isolates selected from eight geographical regions of Ghana and investigated for their technological properties, showed different patterns of growth and flocculation but otherwise similar technological characteristica. Most of the isolates produced pito having sensory attributes, which compared favourably with commercially produced pito. PMID- 16261867 TI - [The results of Lichtenstein operation for groin hernias--prospective multicenter study]. AB - Authors describe the short and long term results of a prospective multicentric Lichtenstein-trial started in March 1999. For the first time in our country, the exchange of information between the participating institutes has been arranged via Internet. 1434 patients were admitted for primary hernia repair and included in the trial. However, only 714 (49.8 per cent) have attended at the last follow up control 4 years after surgery. Premilene and Prolene meshes were used in the course of Lichtenstein's tension-free procedure. RESULTS: Wound suppuration occurred only in a very small number of the patients (0.76 per cent) and there was merely one occasion when the mesh implant had to be removed because of sepsis. The intensity of postoperative pain and demand for analgesics was very low. During the follow-up period of 4 years 16 recurrent hernias were diagnosed (2.4 per cent). A hydrocele developed in 8 patients (0.5 per cent) while necrosis of the testicle was found in 4 patients (0.27 per cent). We realised that both the attitude and practice of the institutes participating in the study has changed thoroughly and the Bassini-repair seems to be replaced by the Lichtenstein procedure in these surgical departments. CONCLUSION: The study has reached its objective in more aspects: First it helped to achieve a significant reduction in recurrence rates and it also has shown that there is no increase in wound suppuration. The great number of surgeons performing this kind of inguinal hernia repair and their good results also indicate that this operation in less depending on surgical skills and practice than the Bassini-repair. The study has largely contributed to the national spread of this operation and has doubtlessly verified the advantage of information and data processing through the Internet, ie. a prompt and correct evaluation of the data recorded PMID- 16261868 TI - [Response rates following neoadjuvant chemotherapy and breast preserving treatment in patients with locally advanced breast cancer]. AB - INTRODUCTION: The neoadjuvant chemotherapy is increasingly being used in the treatment of patients with locally advanced breast cancer. We describe the hypothesis of the biological behaviour of breast cancer supporting the reason for the existence of this treatment. The improvement of neoadjuvant chemotherapy is being discussed as well as the advantages, disadvantages and problems of the treatment. THE AIM OF EXAMINATION: To study the results of neoadjuvant chemotherapy in patients with locally advanced breast cancer and the proportion of breast preserving surgery after the treatment. METHODS: Sixty seven patients were given neoadjuvant chemotherapy treatment between 01.01.1999 and 12.31.2003. Twenty three patients were stage III A while 35 stage III B and 9 stage III C. 63% of the patients received CEF chemotherapy and 19% were given MMM. 18% were given neoadjuvant Taxotere + Carboplatin and 4% were given Taxotere + Farmorubicin chemotherapy. RESULTS: After neoadjuvant chemotherapy 5 patients had SD (stable disease), 32 patients had MR (minor response) and in 28 cases patients had PR (partial response). Two patients showed pCR (complete pathologic response). Twenty patients (30%) had breast preserving surgery. CONCLUSIONS: On the basis of our own experience neoadjuvant therapy is justified in patients with locally advanced breast cancer as they have bigger chance for breast preserving surgery. If mastectomy and axillary block dissection has to be carried out they are easier to perform. Taxans must be introduced for neoadjuvant treatment in order to improve our results. A longer follow-up is necessary before drawing final conclusions. PMID- 16261869 TI - [Covering of bronchial stump fistula with omental flap]. AB - Previous lobectomy was performed on a 71 year old woman because of right inferior lobe lung adenocarcinoma. On the 25th postoperative day bronchial stump leak developed. During reoperation authors covered the fistula with a gastroepiploic omental flap which was pulled through an incision on the diaphragm. The empyema and air leak of the stump disappeared, there were no abdominal complications. This technique was used the first one in Hungary in this patient. PMID- 16261870 TI - [Inflammatory fibroid polyp of the ileum causing intestinal invagination]. AB - Inflammatory fibroid polyps are rare benign tumor-like lesions of the gastrointestinal tract. Most frequently they are localized in the gastric antrum but can develop anywhere in the GI tract. In the small intestine the ileum is the most common site, where these polyps can cause invagination and intussusception. This happened to the patient we describe here. The latest immunohistochemical, ultrastructural and molecular genetic studies are suggestive of dendritic cell origin of inflammatory fibroid polyps, with myofibroblastic differentiation in some cases. PMID- 16261871 TI - [Late metastases of cutaneous malignant melanoma on the abdominal wall to the small and large bowel]. AB - We describe the case of a 56 years old man, who was operated on with abdominal wall skin malignant melanoma 5 years ago. He received postoperative DTIC + Intron A treatment. Five years later he presented with complaints of epigastric pain, melena, hematochezia, anorexia and fatigue. Upper gastrointestinal tract endoscopy showed a tumour mass in the duodeno-jejunal flexure and colonoscopy showed a tumour in the large bowel. Histology verified anaplastic carcinoma. The patient was operated on. We found metastases in the small and the large bowel The patient underwent resection of the jejunum and right hemicolectomy. We describe the different types of metastases of malignant melanomas symptoms, therapies and prognosis. PMID- 16261872 TI - [The effects of classic and delayed ischemic preconditioning on the oxidative stress in small bowel autotransplantation model]. AB - Ischemic Preconditioning (IPC) can reduce ischemia/reperfusion injury produced during small bowel transplantation. We investigated the effects of classic and delayed preconditioning on oxidative stress prior to autotransplantation. Total orthotopic intestinal autotransplantation was performed on 18 dogs. In group I (G I, non-preconditioned) 3-hours cold preservation in University of Wisconsin solution followed by 1-hour of reperfusion. In group II (G-II, classic preconditioned) before this procedure the intestine was preconditioned with 4 cycles of 5 minutes ischemia and 10 minutes reperfusion (IPC protocol). In group III (G-III, delayed preconditioned) on day 1 the animals underwent IPC protocol, and on day 2 autotransplantation was performed. We determined the content of malondialdehyde (MDA), reduced glutathione (GSH), and the activity of superoxide dismutase (SOD) in tissue samples. Our results showed increased lipid peroxidation, slightly elevated GSH level and decreased SOD activity in G-I. In G II MDA slightly elevated, GSH increased markedly and SOD activity preserved by the end of reperfusion. In G-III GSH significantly increased and SOD activity passed the control activity. Our findings confirmed that both forms of preconditioning could moderate the severity of oxidative stress prior to preservation and auto transplantation. Delayed preconditioning is more effective especially to protect bowel tissue against oxidative injury. PMID- 16261873 TI - [Current concerns in surgery]. PMID- 16261874 TI - [Cerebral hemorrhagic stroke and gastric myoelectric activity. Case report]. AB - We showed case of subarachnoid hemorrhage with massive intracerebral hematoma of the left hemisphere and associating gastric myoelectrical disturbances. The cerebral angiography did not show vascular malformation. The electrogastrogram (EGG) at 2 days after the onset of neurological symptoms revealed tachygastria up to 66.7% of the recording time. PMID- 16261875 TI - [Early disturbances of gastric myoelectrical activity in a patient with brainstem hematoma. Case report]. AB - Described early disturbances of gastric myoelectrical activity in 18-years old patient, which appeared immediately after craniocerebral trauma. Brain MRI in a patient showed brainstem hematoma. Bradygastria amounts to 53.6% of the recording time. PMID- 16261876 TI - Cardiac syndrome X--autonomic system disorders. AB - BACKGROUND: The abnormal neural regulation of the heart is one of the most important pathogenesis of cardiac syndrome X (CSX). The aim the study was to evaluate changes in autonomic nervous system activity (ANS) in CSX patients. METHODS: Forty-two patients (age 46 +/- 9 years) with CSX and forty-two healthy volunteers (age 47 +/- 9 years) as a control group were included in the study. Function of ANS was estimated with non-invasive tests based on simple cardiovascular reflexes. RESULTS: In 50% of CSX patients was diminished response to parasympathetic stimulation in deep breathing test. In response to sympathetic stimulation (the changes in body position, hand grip test) was observed decreased 30/15 ratios. The cardiac autonomic activities were altered with parasympathetic withdrawal and/or sympathetic abnormality in patients with CSX. CONCLUSIONS: The population of CSX patients is heterogeneous with normal autonomic system (about 23% of population) and abnormal ANS activity (about 77% patients). In cardiac syndrome X patients selected tests of ANS indicated presence of disturbances in sympathetic-parasympathetic balance. PMID- 16261877 TI - [Ventilatory efficiency during incremental exercise in relation to various types of myosin heavy chain content in vastus lateralis m. quadricipitis femoris in healthy young men]. AB - The main aim of this study was to determine whether ventilatory efficiency during incremental exercise is related to various types of myosin heavy chain (MyHC) content in vastus lateralis in. quadricipitis femoris in young healthy men. The respiratory efficiency was expressed by VE/VCO2 slope, determined during sublactate threshold power outputs of the incremental exercise. A significant negative correlation between V(E)/VCO2 and P(ET)CO2 (amounting to: at rest r = 0.43 p < 0.05; at the LT r = -0.87 p < 0.01; at the VO2 max r = -0.95 p < 0.01) was observed. Moreover, the level of PETCO2 at rest and during exercise was not related to the MyHC content of the vastus lateralis muscle. No relationship between the content of various types of MyHC and respiratory efficiency was found. PMID- 16261879 TI - Effects of vagal pacing on food intake and body mass in pigs. AB - In obesity patients inexpensive, non-invasive, low risk treatment remains a holy grail. The purpose of this study was to evaluate effect of long term and low frequency vagal pacing on feeding behavior in pigs. METHODS: Two groups of animals were investigated, first control with sham operation and second group with microchip (MC) on both vagal nerves placed laparoscopically. In both groups EGG was performed before and after MC implantation. Parameters of stimulation were stable (amplitude 170 mV, frequency 1 Hz, impulse duration 170 ms). RESULTS: MC group demonstrated continuous decrease in body weight gain during 8 weeks of experiment at average of 3.73 +/- 0.5 vs. 4.83 +/- 1.1 (p < 0.05). Food intake also decreased in MC group and was 16.38 +/- 1.3 vs. control 17.5 +/- 1.7 kg/w (ns). EGG recording showed decreased percent normogastria in MC group 26.66 +/- 10 vs. 76.2 +/- 16 (p < 0.05) mostly at cost of tachygastria 62.3 +/- 15 vs. 35.4 +/- 16% of the recording time. CONCLUSION: MC vagal pacing mostly decreased body weight being without significant influence on food intake causing gastric dysrhythmia. PMID- 16261878 TI - Effect of bowel decontamination with metronidazole and vancomycin on gastroduodenal myoelectric activity. AB - It is well recognized that prolonged antibiotic therapy leading to gut decontamination often results in side effects and may lead to colonization of gut with pathologic bacteria. Changes of a gut microflora could play a role in dysmotility of gastrointestinal tract. The aim of the study was to evaluate influence of intraluminal colon anaerobic and aerobic bacterial flora on myoelectric activity of duodenum and stomach. A myoelectric activity recordings using electrodes implanted on small bowel of the conscious rats were performed. Group I was scheduled for control recording, group II for recordings in 4th day after metronidazole (M) administration (30 mg/kg) and group III for recordings after vancomycin (V) administration (15 mg/kg) respectively. Rat's stools were cultured for confirmation of changes in colon flora composition. Recordings were previously filtered digitally with bandwidth filter 0.01-0.1 Hz and 0.1-1.0 Hz to extract gastric and duodenal slow wave respectively and than analyzed with Fast Fourier Transformation. Baseline duodenal slow wave frequency in control group revealed 0.60 +/- 0.05 Hz. M increased slow waves frequency to 0.64 +/- 0.13 Hz and V did not 0.58 +/- 0.09 Hz (p > 0.05). Slow wave dominant frequency of the stomach showed decrease of frequency from control 0.035 +/- 0.04 to 0.025 +/- 0.06 Hz after M (p < 0.05). Pretreatment with V also did not influence slow wave dominant frequency in comparison to control group (0.036 +/- 0.07 Hz, p > 0.05). CONCLUSION: Only pretreatment with M significantly decreased gastric slow wave frequency. One can speculate that M effects are related not only to gut decontamination but also directly affects ENS. We propose hypothesis that M influence on slow wave frequency may be related not only to its antimicrobial activity but to its potential neurotoxic action on intramural ENS neurons. PMID- 16261880 TI - Anorectal and urinary dysfunction after surgery for rectal cancer. AB - AIM: Despite several studies conducted, the relationship between anorectal and urinary dysfunction has not been ultimately determined. METHODS: Forty four patients have been included into the study and divided into three groups according to the type of procedure: Group A - abdomino-perineal resection, group B - low anterior resection and group C - local excision. The urodynamic workup performed 5-9 months after surgical procedure consisted of uroflowmetry, residual urine volume measurement, and cystometry. The following urodynamic parameters were evaluated: sensory threshold, maximal urethral flow within first 30% of voiding time and during the first 5 seconds. Residual volume was controlled by abdominal ultrasound. Anal pressure profile was recorded for evaluation of resting and squeeze anal pressure, length of anal high-pressure zone, radial asymmetry and vector volume. Patients were asked to fulfil a standard questionnaire of voiding dysfunction and fecal continence. RESULTS: Mean values for groups A, B and C revealed for anorectal symptoms 4.7 (SD 3.4), unaffordable symptoms 3.0 (SD 3.3), and for urinary symptoms 2.5 (SD 2.0), 2.9 (SD 1.9), 0.2 (SD 0.6), respectively. Coexistence and aggravation of anorectal and/or urinary symptoms revealed high correspondence within group B (p = 0.055), while this kind of correlation was not observed in group C. Urinary disturbances suggested in uroflowmetric study were the most common in group B (90%). The percentage of patients with manometrically confirmed dysfunction after low anterior rectal resection was twice the value, as compared to the percentage of patients after local excision (p < 0.05). CONCLUSIONS: Our data revealed significant correlation between anorectal and urinary dysfunction after rectal surgery and suggests partially similar pathomechanism of these malfunctions, mediated by direct damage to pelvic autonomic plexus during extended rectal dissection for oncological reasons. PMID- 16261881 TI - Mast cell and eosinophil infiltration in various types of solid malignant tumors. AB - 50 biopsy samples of solid malignant tumors were studied for the presence and numbers of mast cells and eosinophils in the tumor parenchyma and tumors' stromata. Eosinophil infiltrations were very abundant at sites where hemosiderin deposits were present. It is probable that the eosinophils infiltration occurs as a response to the presence of intraerythrocytic facultative pathogens that oxidize hemoglobin and cause its conversion into hemosiderin. PMID- 16261882 TI - Comparing the sensitivity of anaplasma phagocytophilum DNA detection in Ixodes ricinus ticks by amplifying a fragment of the epank-1 and the 16s rDNA genes. AB - The study compared the sensitivity of two genetic markers (epank1, 16S rDNA) in identifying anaplasma DNA by using the polymerase chain reaction (PCR) technique. Material for anaplasma DNA studies was isolated from I. ricinus. Amplification of the 16S rDNA gene resulted in 5.77% positive samples, the value for the epank-1 gene equaled 11.54%. The results suggest that the sensitivity of the reaction for the epank-1 gene is greater than that of 16S rDNA for the identification of anaplasma DNA from isolates of I. ricinus. The application of epank- in identifying A. phagocytophilum DNA can change the pattern of infection in these arachnids in a given area. The results obtained in this study and cared out by another authors point at small useful a conservative gene as a marker for diagnostic goals. PMID- 16261883 TI - Reassessment of defibrase in treatment of acute cerebral infarction: a multicenter, randomized, double-blind, placebo-controlled trial. AB - OBJECTIVE: To evaluate the efficacy and safety of defibrase in patients with acute cerebral infarction by a large sample, multicenter, randomized, double blind, placebo-controlled clinical trial. METHODS: Patients with acute cerebral infarction within 12 hours of stroke onset were randomly assigned to receive either an initial intravenous infusion of defibrase 15 U plus normal saline 250 mL or 250 mL of normal saline only. Subsequent infusions of defibrase 5 U or placebo (normal saline) were given on the 3rd, 5th, 7th, and 9th day, respectively. Both groups received standard care of acute cerebral infarction. The primary efficacy outcome was functional status (Barthel Index) at 3 months after treatment. Safety outcome were bleeding events and mortality rate. Secondary outcome included Chinese Stroke Scale (CSS) score at 14 days and recurrence rate of stroke at 1 year. RESULTS: A total of 1053 patients were enrolled at 46 centers from September 2001 to July 2003, and 527 patients were randomly assigned to receive defibrase and 526 to receive placebo. A similar proportion of patients in both groups completed a full course of treatment. There was a significantly greater proportion of favorable functional status (Barthel Index > or = 95) in defibrase group than in placebo group at 3 months (52.2% vs. 42.8%, P < 0.01), and the proportion of dependent functional status (Barthel Index < or = 60) was a little lower in defibrase group compared with placebo group (27.7% vs. 32.4%). These differences were more obvious among patients who were treated within 6 hours of stroke onset. Patients in defibrase group had better improvement with respect to CSS score than those in placebo group at 14 days (P < 0.05). Recurrence rate of stroke at 1 year was lower in the defibrase group compared with placebo group (6.2% vs. 10.1%, P = 0.053). Patients in defibrase group had higher risk of extracranial bleeding events (4.7% vs. 1.5%, P < 0.01) and a tendency of higher risk of symptomatic intracranial hemorrhage. The hemorrhage incidence was higher in patients with fibrinogen level < 130 mg/dL than > or = 130 mg/dL (10.6% vs. 3.8%, P < 0.05). Mortality rate at 3 months were slightly higher in defibrase group than placebo group (5.9% vs. 4.2%). CONCLUSIONS: The defibrase is effective to improve neurological function and function of daily living for patients with acute cerebral infarction within 12 hours of symptom onset. The efficacy was even better for acute cerebral infarction within 6 hours of onset. The increased risks of intra- and extracranial hemorrhage during defibrase administration were related to the plasma fibrinogen level. PMID- 16261884 TI - A pilot trial for severe, refractory systemic autoimmune disease with stem cell transplantation. AB - OBJECTIVE: To evaluate the feasibility, efficacy, and safety of high dose immunosuppressive therapy (HDIT) and autologous hemopoietic stem cell transplantation (HSCT) with CD34+ cell selection in patients with severe, refractory autoimmune diseases. METHODS: Twenty-six patients with persistent systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), primary Sjogren's syndrome (pSS), or systemic sclerosis (SSc) who had been treated unsuccessfully with conventional treatment were enrolled in the trial in Peking Union Medical College Hospital from September 1999 to June 2004. The patients received HDIT with 200 mg/kg cyclophosphamide followed by an infusion of autologous stem cells that were CD34 selected. Disease activity, adverse effect, hemopoietic and immune reconstitution, and time to recurrence of disease were monitored. RESULTS: Overall treatment related mortality was 7.7% (2/26) with 1 patient died of cytomegalovirus infection and another of severe pneumonia. Relapse occurred in 3 SLE patients (17.6%) in 37, 26, and 19 months posttransplantation respectively, and 1 RA patient in 15 months posttransplantation. SLE Disease Activity Index (SLEDAI) scores of SLE survivors decreased significantly (P < 0.01). RA patients recorded a drop of Disease Activity Score 28 (DAS 28). The pSS patient remained symptoms free up to now, more than 50 months after the transplantation. CONCLUSION: HSCT can be performed relative safely in patients with severe autoimmune disease. Short-term effect of HSCT is promising. However treatment related mortality and relapse were observed in a subset of patients. PMID- 16261885 TI - Variation of protein's expression correlated dto the drug resistance after sequential anti-cancer treatment in human lung cancer cell line. PMID- 16261886 TI - Association between pemphigus vulgaris and human leukocyte antigen in Han nation of northeast China. AB - OBJECTIVE: To investigate the relationship between pemphigus vulgaris (PV) and human leukocyte antigen (HLA) in Han nation of northeast China. METHODS: Standard microcytotoxicity test and polymerase chain reaction-sequence specific primers method were used to detect the HLA class I antigens and HLA-DRB1 and DQB1 alleles in 27 patients with PV and results were compared with control group. RESULTS: Gene and phenotype frequencies of HLA-A3, A26(10), B60(40), and B13 (27.99%, 48%; 16.11%, 30%; 23.02%, 41%; 16.11%, 30%, respectively) increased significantly in PV group compared with control (1.01%, 2%; 0.5%, 1%; 4.61%, 9%; 5.13%, 10%, respectively). After P value correction, the difference of A3, A26 (10), and B60 (40) between the two groups was still significant. The gene frequencies of HLA DRB1*140x (1401, 1404, 1405, 1407, 1408), DRB1*120x, and DQB1*0503 alleles in PV group (42.26%, 25.46%, and 23.02%) were significantly higher than control group (5.09%, 7.74%, and 1.89%). After P value correction, the difference was still significant between the two groups. CONCLUSION: PV significantly relates with HLA in PV patients of Han nation of northeast China. PMID- 16261887 TI - Using of multislice helical CT colonography in patients with malignant lesions of colon. AB - OBJECTIVE: To investigate the techniques and clinical applications of multislice helical computed tomography (CT) colonography in colonic lesions. METHODS: Fifty nine patients with malignant lesions of colon underwent volume scanning using multislice helical CT. Four types of reconstruction including CT virtual colonoscopy (CTVC), shaded surface display (SSD), Raysum, and multiple planar reconstruction (MPR) were used for image post-processing. The results were compared with those of colonoscopy and pathology. RESULTS: Multislice helical CT colonography detected 54 colorectal carcinomas, 4 adenomas with focal carcinoma, 1 non-Hodgkin's lymphoma (NHL). The lesions' number, size, location, morphology, stricture of intestinal cavity, infiltration, and metastasis were shown satisfactorily by multislice helical CT colonography. Whole colon could be shown in all patients. CT colonography displayed 4 synchronous colonic tumors, 1 ascending colon carcinoma combined with left renal carcinoma among 54 patients with colonic carcinomas. The accuracy of location of CT colonography was 100%. There were 9 cases that CT showed the tumor location was different from the finding of conventional colonoscopy, while all of the CT location were proven exact by operation. CT colonography also displayed the infiltration of serous layer and fatty tissue in 45 cases; 21 cases matched the pathological results in all the 24 cases of suspicious lymph node metastasis, the sensitivity was 87.5%, the specificity was 90.6%; 9 cases hepatic metastasis, 2 ovarian metastasis, and 1 double adrenal gland metastasis. CONCLUSIONS: Multislice helical CT colonography is effective in preoperative diagnosis, location, stage, and making treatment plan of colorectal carcinoma. It can display the portion not seen during colonoscopy and may have an adjunctive role. PMID- 16261889 TI - MUC5AC expression up-regulation goblet cell hyperplasia in the airway of patients with chronic obstructive pulmonary disease. AB - OBJECTIVE: To determine the number of goblet cells, the change of MUC5AC expression in chronic obstructive pulmonary disease (COPD) patients and the relationship of smoking with goblet cell, MUC5AC, and lung function. METHODS: Eighteen patients undergoing lung resections for a solitary peripheral carcinoma were classified by lung function as having COPD. Twenty patients with normal lung function served as the control group. Normal lobe bronchioles far away from the lesion site were taken for paraffin section. Goblet cells were identified by AB/PAS staining and the expression of MUC5AC in the paraffin's section was tested by immunohistochemistry. RESULTS: Goblet cell hyperplasia was observed in the COPD group. The positive rate of goblet cell in COPD group (0.20% +/- 0.10%) was significantly higher than that in the normal lung function group (0.13% +/- 0.06%, P < 0.05). The positive rate of MUC5AC expression in the COPD group (0.27% +/- 0.09%) was higher than that in the normal lung function group (0.20% +/- 0.10%, P < 0.05). The positive rate of goblet cell in smokers (27.93% +/- 9.00%) of the COPD group and normal lung function group was higher than that in non smokers (17.70% +/- 9.37%, P < 0.05), while MUC5AC expression had no significant difference between smokers and non-smokers (17.88% +/- 6.44% and 10.88% +/- 7.10%, respectively). CONCLUSION: For COPD patients with declined lung function, there were goblet cell hyperplasia and increased expression of MUC5AC. MUC5AC expression up-regulation may due to goblet cell hyperplasia. Smoking may be an important factor for goblet cell hyperplasia. PMID- 16261888 TI - Effect of erythromycin on gastric dysmotility and neuroendocrine peptides in rats with diabetes mellitus. AB - OBJECTIVE: To investigate the effect of erythromycin on the contractive activity of the isolated gastric antrum smooth muscle and somatostatin (SS), vasoactive intestinal peptide (VIP), motilin (MTL), and substance P (SP) in plasma and isolated gastric antrum tissue of diabetes mellitus (DM) rat models. METHODS: Thirty male Sprague-Dawley rats were divided into three groups: control group (n = 10), DM group (n = 10), and erythromycin group (DM models with erythromycin treatment, n = 10). A single dose of streptozotocin (100 mg/kg, dissolved in 0.1 mol/L citric acid buffer, pH4.5) was injected intraperitoneally. After 48 to 72 hours, rats with blood glucose above 16.7 mmol/L and urine glucose level to be (+++) to (++++) over one week were considered successful DM models. The resting tension, mean contractile amplitude and frequency of spontaneous change in isolated longitudinal and circular gastric antrum smooth muscle strips were measured. SS, VIP, MTL, and SP levels in plasma and gastric antrum tissue were measured using radioimmunoassay. RESULTS: (1) In the isolated gastric antrum smooth muscle strips, the gastric motility parameters were lower in DM group than those in control group except circular smooth muscle contractile amplitude and longitudinal smooth muscle contractile frequency. The gastric motility parameters were significantly strengthened in erythromycin group, compared with DM group except longitudinal smooth muscle resting tension (P < 0.01). (2) Plasma SS, VIP, and MTL concentrations in DM group were higher than those in control (P < 0.05), while the SP level decreased (P < 0.05). In the gastric antrum, SS of DM group was significantly higher than that of control group (P < 0.01), while SP and MTL levels were lower than those of control group (P < 0.05 and P < 0.01, respectively). However, the level of VIP in gastric antrum tissue did not change among three groups. The plasma level of SS in erythromycin group was higher than that of DM group (P < 0.05). (3) The blood glucose was lower in erythromycin group than DM group (P < 0.01). CONCLUSIONS: Erythromycin has direct effects on contractive activity of gastric smooth muscle in diabetic rats, but there are few effects on neuroendocrine peptides. Gastric-motility disorders in diabetic rats have a correlation with the changes of neuroendocrine peptide levels in plasma and gastric antrum tissue. PMID- 16261890 TI - Identification of differential genes in ovarian cancer using representational difference analysis of cDNA. AB - OBJECTIVE: To identify differential genes between normal ovarian epithelium tissue and ovarian epithelial cancer using representational difference analysis of cDNA (cDNA-RDA). METHODS: cDNA-RDA was performed to identify the differentially expressed sequences between cDNAs from cancer tissue and cDNAs from normal ovarian tissue in the same patient who was in the early stage of ovarian serous cystadenocarcinoma. These differentially expressed fragments were cloned and analyzed, then sequenced and compared with known genes. RESULTS: Three differentially expressed cDNA fragments were isolated using cDNA from normal ovarian tissue as tester and cDNA from cancer tissue as driver amplicon by cDNA RDA. DP III-1 and DP III-2 cDNA clone showed significant homology to the cDNA of alpha actin gene; DP III-3 cDNA clone showed significant homology to the cDNA of transgelin gene. CONCLUSION: cDNA-RDA can be used to sensitively identify the differentially expressed genes in ovarian serous cystadenocarcinoma. Ovarian serous cystadenocarcinoma involves alteration of multiple genes. PMID- 16261891 TI - Evaluation of international classification criteria (2002) for primary Sjogren's syndrome in Chinese patients. AB - OBJECTIVE: To evaluate the sensitivity and specificity of international classification criteria (2002) for primary Sjogren's syndrome (pSS) and the role of lower lip biopsy in diagnosis of pSS in Chinese patients. METHODS: Patients who were diagnosed by the experts/rheumatologists as pSS during 1990-2002 from the Department of Rheumatology, Peking Union Medical College Hospital were retrospectively collected as experimental group. Patients who were diagnosed as non-pSS connective tissue diseases or non-connective tissue diseases served as control group. Those with a history of head-neck radiation, hepatitis C virus infection, AIDS, lymphoma, sarcoidosis, graft versus host disease (GVHD), and anti-acetylcholine drug use were exempted. Both groups were required to complete questionnaires about symptoms such as dry eyes and dry mouth, and complete the objective tests of keratoconjunctivitis and xerostomia including Schirmer test, corneal staining, unstimulated salivary flow, sialography, lower lip biopsy, and antinuclear antibodies (including anti-SSA/SSB antibodies) test. RESULTS: A total of 330 pSS patients were included in experimental group and 185 non-pSS patients in control group. The mean age of both groups matched (47.8 +/- 10.9 years vs. 46.2 +/- 13.6 years, P > 0.05). The sensitivities of the criteria in pSS patients with lower lip biopsy and in pSS patients without lower lip biopsy were 89.2% and 87.2%, respectively; the overall sensitivity was 88.5%. The specificity was 97.3%. A total of 11.3% pSS patients with negative anti-SSA/SSB anti- bodies were diagnosed as pSS by lower lip biopsy. CONCLUSION: The international classification criteria (2002) for pSS is feasible in Chinese patients. It has high sensitivity and specificity, and may serve as diagnosis criteria in routine clinical practice. PMID- 16261892 TI - Thromboendarterectomy for chronic pulmonary thromboembolism. AB - OBJECTIVE: To evaluate the improving reliability and safety of thromboendarterectomy and perioperative management for chronic pulmonary thromboembolism. METHODS: The clinical data of 12 cases with chronic pulmonary thromboembolism, who underwent thromboendarterectomy assisted by low flow or circulation arrest with deep hypothermia, were reviewed retrospectively. RESULTS: Pulmonary artery pressure decreased 20 to 40 mmHg immediately after surgical procedures in 9 cases. The postoperative pulmonary edema at various degrees happened in 12 cases, among them, 1 died of severe lung infection and pulmonary re-embolism at 19 days postoperation. Computed tomography pulmonary angiography and angiography of 11 cases indicated that the original obstruction of pulmonary artery disappeared. During the follow-up period of 2 months to 5 years, the clinical symptoms and activity was improved. CONCLUSION: Thromboendarterectomy is an effective treatment for chronic pulmonary thromboembolism. The outcome of the surgical procedure needs to be further investigated and followed up regularly according to an evaluative system, because it might be influenced by multiple factors. PMID- 16261893 TI - Reconstruction of the sellar floor following transsphenoidal surgery using gelatin foam and fibrin glue. AB - OBJECTIVE: To introduce a new principle of sellar reconstruction and to evaluate the effectiveness of absorbable gelatin foam and fibrin glue for sellar reconstruction. METHODS: A total of 176 consecutive patients who underwent surgery for pituitary adenomas, cysts, chordomas, or subdiaphragmatic craniopharyngiomas in the sella turcica between January 2001 and April 2003 at Peking Union Medical College Hospital were enrolled. Different techniques of sellar closure and indications for each specific condition were retrospectively reviewed. RESULTS: Seventy-seven (43.7%) patients developed a visible cerebrospinal fluid (CSF) leakage during surgery. Intra-operative CSF leakage were repaired simply with gelatin foam and fibrin glue in 62 (35.2%) patients, and with autologous fat graft and sellar floor reconstruction in 15 (8.5%) patients. Postoperative CSF rhinorrhea occurred only in 1 case. There were no visual deterioration, allergic rhinitis, meningitis, pneumocranium, granulomas, or other complications associated with the reconstruction procedure. CONCLUSION: The procedure of using gelatin foam and fibrin glue and principle of cranial base reconstruction is safe and effective in preventing postoperative complications following transsphenoidal surgery. PMID- 16261894 TI - Multivariate analysis of relationships between iodine biological exposure and subclinical thyroid dysfunctions. AB - OBJECTIVE: To assess the relationships between iodine biological exposure and subclinical thyroid dysfunctions. METHODS: The cross-sectional survey was performed to obtain the epidemiologic data of population in three communities with different iodine biological exposure: mild iodine deficiency [median urinary iodine concentration (MUI) of 50-99 microg/L], more than adequate iodine intake (MUI of 200-299 microg/L), and excessive iodine intake (MUI over 300 microg/L). Univariate and multivariate analysis (logistic regression analysis) were used to analyze the risk factors of subclinical hypothyroidism and subclinical hyperthyroidism. RESULTS: Logistic regression analysis with sex and age controlled suggested that more than adequate iodine intake (OR = 3.172, P = 0.0004) and excessive iodine intake (OR = 6.391, P = 0.0001) increased the risk of subclinical hypothyroidism, while excessive iodine intake decreased the risk of subclinical hyperthyroidism (OR = 0.218, P = 0.0001). Logistic regression analysis including interaction of iodine intake and antibodies [thyroid peroxidase antibody (TPOAb) and thyroglobulin antibody (TgAb)] suggested that excessive iodine intake was an independent risk factor of subclinical hypothyroidism (OR = 6.360, P = 0.0001), but independent protect factor of subclinical hyperthyroidism (OR = 0.193, P = 0.0001). More than adequate iodine intake and it's interaction with TgAb increased the risk of subclinical hypothyroidism independently, in addition, it decreased the risk of subclinical hyperthyroidism at the present of TPOAb. CONCLUSION: Both excessive iodine intake and more than adequate iodine intake could increase risk of subclinical hypothyroidism, supplement of iodine should be controlled to ensure MUI within the safe range. PMID- 16261895 TI - Three-dimensional computed tomography-guided radiofrequency trigeminal rhizotomy for treatment of idiopathic trigeminal neuralgia. AB - OBJECTIVE: To evaluate the effectiveness of three-dimensional computed tomography (3D-CT) guided radiofrequency trigeminal rhizotomy (RF-TR) in treatment of idiopathic trigeminal neuralgia (ITN). METHODS: From 1999 to 2001, 18 patients with ITN were treated with percutaneous controlled RF-TR. Intraoperative 3D-CT scanning was performed to guide the trajectory of the puncture. After correction of the needle tip according to the CT scans and stimulation effects, 2 to 5 lesions were made for a duration of 60-90 seconds at a temperature of 60 degrees C to 75 degrees C depending on the pain distribution and the age of patient. RESULTS: The needles located in foramen ovale. Pain alleviated immediately with no serious complication in all patients. The patients were followed up for an average of 31.5 months (range 24-41 months). Acute pain relief was experienced by 17 patients after the procedure, reaching an initial success rate of 94.4%. Early (< 6 months) pain recurrence was observed in 2 patients (11.1%), whereas late (> 6 months) recurrence was reported in 3 patients (16.7%). Thirteen patients had complete pain control, with no need for medication thereafter. Five cases experienced partial pain relief, but required medication at a lower dose than in the preoperative period. CONCLUSION: 3D-CT foramen ovale locations can raise the successful rate of puncture, enhance the safety, and reduce the incidence rate of complication. PMID- 16261896 TI - Diagnostic delay in women with endometriosis. PMID- 16261897 TI - Prevention and treatment of rejection after simultaneous pancreas-kidney transplantation. AB - OBJECTIVE: To explore methods of preventing and reversing rejection after simultaneous pancreas-kidney (SPK) transplantation. METHODS: Seventeen patients underwent SPK transplantation from September 1999 to September 2003 were reviewed retrospectively. Immunosuppression was achieved by a triple drug regimen consisting of cyclosporine, mycophenolate mofteil (MMF), and steroids. Three patients were treated with anti-CD3 monoclone antibody (OKT3, 5 mg x d(-1)) for induction therapy for a mean period of 5-7 days. One patients received IL-2 receptor antibodies (daclizumab) in a dose of 1 mg x kg(-1) on the day of transplant and the 5th day posttransplant. One patient was treated with both OKT3 and daclizumab for induction. RESULTS: No primary non-functionality of either kidney or pancreas occurred in this series of transplantations. Function of all the kidney grafts recovered within 2 to 4 days after transplantation. The level of serum creatinine was 94 +/- 11 micromol/L on the 7th day posttransplant. One patient experienced the accelerated rejection, resulting in the resection of the pancreas and kidney grafts because of the failure of conservative therapy. The incidence of the first rejection episodes at 3 months was 47.1% (8/17). Only the kidney was involved in 35.3% (6/17); and both the pancreas and kidney were involved in 11.8% (2/17). All these patients received a high-dose pulse of methylprednisone (0.5 g x d(-1)) for 3 days. OKT3 (0.5 mg x d(-1)) was administered for 7-10 days in two patients with both renal and pancreas rejection. All the grafts were successfully rescued. CONCLUSION: Rejection, particularly acute rejection, is the major cause influencing graft function in SPK transplantation. Monitoring renal function and pancreas exocrine secretion, and reasonable application of immunosuppressants play important roles in the diagnosis and treatment of rejection. PMID- 16261899 TI - Expression of a telomerase-associated gene in normal, atrophic, and tumorous testes. AB - OBJECTIVE: To evaluate the expression of telomerase transcriptional elements interacting factor (TEIF) in human testis under different status and its relation with human telomerase reverse transcriptase (hTERT) expression. METHODS: Specific antisera against TEIF were generated by immunization of rabbits with purified recombinated partial TEIF. Samples were assigned to three groups according to their pathological types, including 16 normal testes, 8 atrophic testes, and 6 testicular seminomas. They were subjected to immunohistochemical staining of TEIF and hTERT. Results from both TEIF and hTERT were analyzed semi-quantitatively and compared. RESULTS: The expressions of TEIF and hTERT were detected in all samples of normal, atrophic testes, and seminomas. No differences of TEIF expressions among these three groups were observed (P > 0.05). On the contrary, the expressions of hTERT were significantly lower in atrophic testes compared with those of normal testes and seminomas (both P < 0.05). Nevertheless, co expressions of TEIF with hTERT were revealed to be in normal and malignant cases (P < 0.05) but not in atrophic testes, which generally presented TEIF expression. The cellular distributions of both proteins were similar and mainly in spermatocytes and some Sertoli cells, while were all negative in the interstitial cells and other stromal cells. Conclusions The uniform expressions of TEIF in all these specimens suggest that it may be a marker of testis and its related diseases. The strong expression of hTERT in normal testes and testicular seminomas comparing with the low expression in atrophic testes may suggest a role for telomerase in maintaining proliferation of germ cells. PMID- 16261898 TI - Expression and significance of Survivin mRNA in lung cancer tissue microarray detected by FISH. AB - OBJECTIVE: To investigate the expression of Survivin mRNA in lung cancer tissue microarray (TMA) by fluorescence in situ hybridization (FISH) method, and determine the role and significance of it in lung cancer genesis and progress. METHODS: The expression of Survivin mRNA was detected by FISH method and TMA technology. Fifty-four cases of lung cancer and 10 cases of normal lung tissue were examined. RESULTS: Survivin mRNA was expressed in 66.7% (36/54) of lung cancer; the positive ratio of lung cancer was significantly higher than that of normal lung tissue (0/10; chi2 = 15.238, P < 0.05). The positive ratio of Survivin mRNA was significantly higher in poor differentiated cancer (20/24, 83.3%) than moderate and well differentiated cancer (16/30, 53.3%; chi2 = 5.40, P < 0.05). The positive ratio of Survivin mRNA was significantly higher in group with lymph node metastasis (27/32, 84.4%) than without lymph node metastasis (9/22, 40.9%; chi2 = 11.084, P < 0.05). The positive ratio of Survivin mRNA was significantly higher in stage III-IV(12/13, 92.3%) than stage I - II (24/41, 58.5%; chi2 = 5.066, P < 0.05). CONCLUSION: Survivin mRNA highly expresses in lung cancer, which is related to the progress and malignant behavior. Survivin may play a promoting role in lung cancer genesis and progress and provide a basis for estimating prognosis and treatment. PMID- 16261900 TI - Development on adjustable calibration marker for shock wave focus. PMID- 16261901 TI - Concentrations of nitric oxide in rat brain tissues after diffuse brain injury and neuroprotection by the selective inducible nitric oxide synthase inhibitor aminoguanidine. PMID- 16261902 TI - Less is not more. PMID- 16261903 TI - Remember the children. PMID- 16261904 TI - Tooth avulsion and replantation. PMID- 16261905 TI - Creating smiles in West Africa. PMID- 16261906 TI - Older adults--implications for private dental practitioners. AB - Currently, 35 million people are over the age of 65 in the United States. This number is expected to double to 70 million by 2030 (Figure 1). In California, 3.7 million people are over the age of 65, and this number is expected to increase to 6.4 million in the next 20 years or within the practice lifetime of students presently enrolled in California's dental hygiene and dental schools. The oldest old, those over age 85, are the fastest-growing segment of the United States and California's population. California's aging population will reflect the diversity of the state in general. Table 1 lists California's 65-plus population by age and ethnic/racial categories. By 2030, one in five Americans and Californians will be 65 years or older. Women who reach age 65 can expect to live an additional 19 years of life, while men can expect to live an additional 16 years. The gap in life expectancy between men and women is narrowing due to improvements in medical care, preventive health services, and healthier lifestyles. Figures 2-4 show the improvements in life expectancy at birth, age 65, and age 75 for the U.S. population. In the United States, there are an estimated 1.8 million nursing home beds used by 80 percent of the residents over age 65. A report by the U.S. General Accounting Office estimated that 43 percent of all Americans over age 65 will reside in a nursing home at some time in their life. California currently has approximately 100,000 residents living in one of the 1503 nursing home facilities throughout the state. Nursing home care in California accounts for 5.6 billion dollars. In 1998, the U.S. General Accounting Office reported that one in three California nursing homes was cited for serious or potentially life threatening care problems. With an aging imperative in California, this paper will discuss the implications of an aging society on maintaining oral health throughout one's life, and the ability of dental professionals to meet the oral health needs of this population. PMID- 16261907 TI - The role of safety net providers in delivering oral health services for people with special needs. AB - The U.S. health care system, best suited to acute care for adults, struggles to accommodate vulnerable populations (such as the elderly, disabled, and mentally ill), and struggles even more to find or put in place a system to care for special needs populations. Special needs populations require a protective and preventive system--one that helps families anticipate upcoming needs as patients transition through life cycles, and monitors problems as they arise while coordinating services. Developing such a system, using a life cycle methodology, is a critical health policy frontier. PMID- 16261908 TI - Access to care for people with special needs: role of alternative providers and practice settings. AB - Oral Health in America: A Report of the Surgeon General released in 2000 was the first-ever surgeon general's report on the status of oral health in the United States. It clearly outlined a growing set of challenges in such areas as reducing oral health disparities, improving access to oral and dental care, and prevention of common dental diseases. Findings revealed that 75 percent of dental disease is found in 25 percent of the population. California's children have twice as much untreated decay as their national counterparts. For children with special health care needs seeing a dentist, the data is sparse but a survey of general dentists conducted in 2001 showed that only 10 percent see these children often or very often. PMID- 16261909 TI - Education of dentists in the treatment of patients with special needs. AB - The dental education system has been suggested as the vital link in providing a workforce capable of improving oral health for people with special needs. Dental education institutions not only train dental professionals for their role in providing oral health services for people with special needs, they also provide a significant amount of services to this population in their clinical environments. However, there is no consensus about whether to concentrate the educational efforts on the preor postdoctoral level, or both. Furthermore, it is not clear if educational initiatives in the care of patients with special needs will translate into a larger oral health workforce willing to treat these patients. However, for the purposes of this paper, it will be assumed that more education and training in special care dentistry will lead to better-educated dentists and the desired result of better access to care for special needs patients. The authors will define special needs patients as those who have a chronic physical, developmental, behavioral, or emotional condition, and who also require health and related services of a type or amount beyond that the general population requires. This paper will describe accreditation issues and discuss the advantages and disadvantages of special care education in pre- and postdoctoral training and beyond. PMID- 16261911 TI - 'Doctor time'--the power begins here. PMID- 16261910 TI - Financing oral health services for people with special needs: projecting national expenditures. AB - Low-income people with disabilities or who are elderly have more dental disease, more missing teeth, and more difficulty obtaining dental care than other members of the general population. These realities lead to untreated infection, increased medical costs and needless suffering for the most vulnerable members of our society. It is critical we provide adequate reimbursement for oral health services in order to avoid the tragic and costly consequences of oral neglect. This article focuses on the financial implications of delivering oral health services to low-income individuals who are "aged, blind, and disabled" in the United States. The experience of providing oral health services in California for these populations is extrapolated to predict the cost implications of a national reimbursement system for ABD adults under Medicaid and reform Medicaid oral health programs for vulnerable children. The new federal dollars required to implement this legislation would be more than offset by a conservatively estimated 0.5 percent reduction in costly emergency room and hospital charges for the treatment of serious dental problems, as well as a reduction in the prevalence and severity of several general health conditions. Treating and/or preventing oral infection and disease for the ABD populations in our country will significantly reduce overall health care costs, improve quality of life, and end needless suffering for America's most vulnerable citizens. Treating and/or preventing oral infection and disease for this population simply is the right thing to do. PMID- 16261912 TI - Combination therapy in the treatment of malaria. PMID- 16261913 TI - Safety and efficacy of artemether-lumefantrine in the treatment of uncomplicated falciparum malaria in Ethiopia. AB - OBJECTIVE: To document baseline data on the efficacy and safety of artemether lumefantrine for the treatment of uncomplicated falciparum malaria in Ethiopia. DESIGN: Patients diagnosed for P. falciparum, who were treated with six doses of artemether-lumefantrine over three days, were followed for 28 days and treatment outcomes classified based on the WHO (2003) protocol. SETTING: Four health facilities located in malarious areas in two regions: Alamata and Humera hospitals in Tigray region and Assendabo and Nazareth in Oromia region. SUBJECTS: Patients with body weight of more than 10 kgs, excluding pregnant women, who or their guardians consented to participate in the study after fulfilling the inclusion criteria were enrolled in the study for a follow-up period of 28 days. MAIN OUTCOME MEASURES: Proportion of treatment success and adverse drug effects that required discontinuation of treatment and/or follow-up. RESULTS: A total of 213 patients who fulfilled the enrolment criteria completed the 28 days follow-up after treatment with artemether-lumefantrine. A treatment success rate of 99.1% (95% confidence interval [CI] 96.9, 99.8) and no adverse effects or complaints related to the drug that required discontinuation of treatment or withdrawal from follow-up was reported. Treatment success was not achieved in 213 (0.9%) subjects for whom fever and peripheral parasitaemia was demonstrated on day 21 and 28. The day 21 and day 28 blood samples of the treatment failure cases were not PCR corrected. CONCLUSION: The artemisinin based combination drug artemether lumefantrine has shown very high (99.1%) clinical and parasitological cure for the treatment of uncomplicated falciparum malaria with no reports of adverse reaction that required withdrawal of treatment or discontinuation of follow-up. In the presence of the low efficacy of sulfadoxine-pyrimethamine, chloroquine and amodiaquine, the use of artemether-lumefantrine for the treatment of uncomplicated falciparum malaria is the best choice for Ethiopia. PMID- 16261914 TI - Efficacy of sulfadoxine-pyrimethamine for the treatment of uncomplicated falciparum malaria in Ethiopia. AB - OBJECTIVE: To assess the status of the therapeutic efficacy of sulfadoxine pyrimethamine for the treatment of uncomplicated falciparum malaria to enable evidence based policy decisions. DESIGN: The study used the new WHO (2003) protocol for the assessment of the therapeutic efficacy of anti-malarial drugs. SETTING: Eleven health facilities located in malarious areas with seasonal transmission. SUBJECTS: Patients aged six months and above who presented to the health facilities for febrile illness and for whom consent was obtained to participate in the study after fulfilling the inclusion criteria were enrolled in the study. MAIN OUTCOME MEASURES: Proportion of treatment failures. RESULTS: In eleven (90.9%) of the sites, where adequate sample was collected, a total of 598 subjects were enrolled and 487 (81.4%) completed the follow-up. A mean treatment failure rate of 35.9% (95% confidence interval [CI] 31.8, 40.3) on the 14 days follow-up and 71.7% (95% CI 67.5, 75.9) on the 28-days follow-up was recorded (not PCR corrected). The mean clinical failure on the 14-days follow-up was 20.9% (95% CI 17.5, 24.7) and 70% (n=10) sites had aggregated clinical failure rates higher than 15%, while in 80% (n=10) sites the total treatment failure exceeded 25%. There was no significant difference in treatment failure rates in areas with malaria transmission duration of six months and above as compared to areas with below six months of transmission (odds ratio [OR] = 0.9, 95% CI 0.43,1.83 p = 0.75). The difference in mean treatment failure between the <5 and > or =15 years of age was not significant (OR 0.8, 95% CI 0.39,1.67 P = 0.54). CONCLUSION: The level of treatment failure detected is much higher than the WHO recommended tolerable levels. The findings, therefore, strongly indicate the need for an immediate review of the existing national anti-malarial treatment guideline. PMID- 16261915 TI - Malaria prevention practices in Mopti region, Mali. AB - OBJECTIVES: To assess insecticide-treated nets (ITNs) use, other malaria prevention measures, knowledge of malaria and diffusion of information about ITNs by the National Malaria Control Programme (NMCP) in Mali. DESIGN: A descriptive cross-sectional study. SETTING: Four villages in Mopti region, Mali that had participated in the Mopti Regional Malaria Control Program (MRMCP) ITN Education Programme five years prior. PARTICIPANTS: Three hundred thirty nine randomly selected households. Within each household, mothers of children one to nine years of age were interviewed regarding knowledge of malaria and prevention practices. RESULTS: Overall, 11% of households used ITNs, with 97% of these in two villages. Ninety eight percent of households used bednets, 22% used insecticide sprays and 39% used mosquito coils. Significant predictors of ITNs use were: head of household literacy, larger family size, Bambara ethnicity, hearing about the NMCP and hearing about ITNs from health agents. Reasons why ITNs were not used included not knowing anything about ITNs, cost and not having net impregnation services readily available in the village. Levels of knowledge concerning malaria disease, transmission and prevention varied amongst the four villages. ITN households had significantly higher levels of knowledge about malaria and its prevention. CONCLUSION: Five years after the implementation of the MRMCP, ITNs use was low and diffusion of malaria prevention information was unequal among villages. Future efforts in improving the programme must recognise these differences in knowledge and ITNs utilisation and make village-specific changes that are acceptable for each village. PMID- 16261916 TI - Impact of emergency mass immunisations on measles control in displaced populations in Gulu district, northern Uganda. AB - OBJECTIVE: To assess the impact of supplemental mass measles immunisations. DESIGN: Retrospective study of hospital and health centre records. SETTING: Gulu district, Northern Uganda, having approximately 81% of the population living in internally displaced persons' (IDPs) camps. RESULTS: The mean age in months for 4,812 measles cases seen was 28.2 +/- 46.0 (p < 0.0001). Supplemental mass immunisations in 1997 and 2000 caused a 91% reduction of measles cases, 93% reduction of mortality, 91% reduction of bed-days and 79% reduction of outpatient cases. There was a 67% reduction in mean measles case admissions, 63% reduction in mean measles mortality, and 73% reduction in mean measles bed-days following district mass measles immunisations in 1997. However, following IDPs camps supplemental immunisations in 2000; there was 82% reduction of mean measles case admissions, 80% reduction of mean measles mortality and 88% reduction of mean measles bed-days. CONCLUSIONS: In similar situations, supplemental mass measles immunisations should be focused on IDPs camps with a wide age group in addition to improved routine immunization activities in the entire district. PMID- 16261917 TI - Capacity of community-based organisations to disseminate sleeping sickness information. AB - OBJECTIVE: To assess the capacity of established community based organisations (CBOs) to disseminate information on sleeping sickness control. DESIGN: Participatory interview process administered to randomly selected CBOs in a tsetse and trypanosomosis endemic area. SETTING: Busia district, Western, Kenya. RESULTS: Community based organisations especially women groups and farmer field schools that are internally initiated have the potential to contribute greatly to sustainable sleeping sickness dissemination strategies. The study indicated a mean reach of between 400-600 persons per day, but with a range of up to 1000 persons per day. CONCLUSION: Internally initiated women groups may be the best options for targeting health education programmes with the aim of ensuring sustained community participation. PMID- 16261918 TI - Comparison of antibiotic use between an 'open' and a 'closed' intensive care unit. AB - OBJECTIVE: To determine differences in antibiotic use between a 'closed' ICU in a hospital in Israel and an 'open' ICU in a Kenyan hospital. DESIGN: Retrospective comparative study. SETTING: The ICU of Beilinson hospital in Rabin Medical Centre, Tel Aviv, Israel and the ICU of Mater Hospital, Nairobi Kenya. SUBJECTS: One hundred and forty patients from Beilinson and one hundred fifty one patients from Mater Hospital were enroled. MAIN OUTCOME MEASURES: Antibiotic use in the different ICU settings. RESULTS: Seven different antibiotics namely Erythromycin, Meropenem, Taxobactam/Piperacillin, Metronidazole, Gentamycin, Ceftriaxone and Cefuroxime were used in sufficient numbers in both centres to allow for statistical analysis. Four of these seven namely metronidazole, gentamycin, ceftriaxone and cefuroxime demonstrated statistically significant difference between the two centres. CONCLUSION: The level, of antibiotic use is demonstrably higher in 'open' ICUs with fewer controls to antibiotic prescriptions than in 'closed' ICUs with stricter control. PMID- 16261919 TI - Ameloblastic fibroma at the University of Nairobi Dental Hospital. AB - OBJECTIVE: To document the occurrence of cases of ameloblastic fibroma. DESIGN: A retrospective study. SETTING: The Division of Oral Pathology and Oral Medicine histopathology laboratory at the Faculty of Dental Sciences, University of Nairobi, January 1991 to December 2000. RESULTS: Seven complete records of histopathologically confirmed cases of ameloblastic fibroma were identified. There were four male and three female cases with an age range of 10 to 22 years (mean=14 years). Remarkably, six of the cases had lesions in the molar regions of the mandible with a single case with a maxillary molar area lesion. One of the cases had the neoplasm associated with an unerupted tooth. From the clinical notes all the patients were managed by meticulous enucleation and curettage of the lesions. After follow up periods ranging from several months to eight years no recurrences had been recorded. CONCLUSION: Evidently ameloblastic fibroma is an uncommon neoplasm as noted in the earlier literature. Although the lesions can be quite extensive at the time of diagnosis, the recurrence rate appears to be relatively low if the neoplasms are meticulously extirpated. PMID- 16261920 TI - Immunoassay and polymerase chain reaction techniques for detection of enterotoxigenic Bacillus cereus. AB - OBJECTIVES: To compare the Reverse Passive Latex Agglutination (RPLA) and Enzyme Linked Immunosorbent Assay (ELISA) techniques with a Polymerase Chain Reaction (PCR) for detection of enterotoxigenic Bacillus cereus. DESIGN: A cross-sectional study. SETTING: The Department of Public Health, Pharmacology and Toxicology, Faculty of Veterinary Medicine, University of Nairobi. SUBJECTS: Forty seven Bacillus cereus strains previously isolated from foods. MAIN OUTCOME MEASURES: Detection of hemolysin BL, non-hemolytic enterotoxin, binding protein gene (hblA) of the hemolysin BL, and binding protein gene (nheA) of nonhemolytic enterotoxin. RESULTS: Twenty five (53.2%) of the isolates produced hemolysin BL, while 81% of them produced non-hemolytic enterotoxin. Thirty eight (38.3%) produced both hemolysin BL and non-hemolytic enterotoxin. A polymerase chain reaction amplification assay detected the presence of hblA gene in all hemolysin BL positive isolates and nheA gene in 91.5% of non-hemolytic enterotoxin positive isolates. There was a strong association between PCR test and RPLA test (Pearson's X2 = 12.65; p < 0.001) as well as between PCR test and visual immunoassay test (Pearson's chi-square X2 = 18.46: p < 0.01). CONCLUSION: Polymerase chain reaction amplification assay technique for detection of enterotoxigenicity of B. cereus compare well with the immunoassay tests. The technique is sensitive detecting even strains with silentgenes, and is rapid with the test complete within 24 hours. PMID- 16261921 TI - Pesticide use on agricultural fields and health problems in various activities. AB - OBJECTIVES: To assess the health status, attitude and level of awareness of safe pesticide handling practices of farm workers engaged in the application of pesticides on agricultural farms. DESIGN: Cross sectional study. SETTING: Two farms in northwest Ethiopia, 2004. SUBJECTS: Farm workers of job categories; sprayers, pest assessors, supervisors, spraymachine mechanics and tractor operators. RESULTS: The farm workers had respiratory symptoms of cough, phlegm and wheezing. Systolic and diastolic blood pressures did not show abnormalities. Liver function tests showed elevated values. Respiratory symptoms in the farm workers revealed that cough phlegm and wheezing at Ayehu farm were significantly (p < 0.05) higher than the controls. Alkaline phosphatase (ALP) at Birr Farm in the sprayers and mechanics were significantly higher than the controls (p < 0.05). The ALP value in the sprayers, glutamate pyruvate transaminase (GPT) in the assessors and glutamate oxaloacetate transaminase (GOT) in the sprayers and mechanics at Ayehu were significantly higher than the controls (p < 0.05). From a total of 82 farm workers 35.7% at Birr and 75% at Ayehu Farm described that they were not formally instructed about safe pesticide handling methods. CONCLUSION: The farm workers health is affected by the unwise use of pesticides. The level of awareness and attitude on safe pesticide handling practices is low. It is recommended that appropriate type of personal protective device (PPD), in service training about the proper use of chemical pesticides and periodic medical check up should be fulfilled to minimise the adverse health effects of chemical pesticides. PMID- 16261922 TI - Congenital malformations in mice induced by addiction to alcohol and cocaine. AB - OBJECTIVE: To study the teratogenic effect of either alcohol alone, cocaine alone, or a combination of both alcohol and cocaine on mice foetuses. DESIGN: Eighty pregnant mice were divided into four equal groups. In the first (alcohol) group, the pregnant females were given absolute ethanol at 2.5 gm/100 gm twice daily by oro-gastric incubation. In the second (cocaine) group, the pregnant females received cocaine hydrochloride at 20 mg/ 100 gm daily by intraperitoneal injection. In the third (combined) group, the pregnant females received alcohol at 2.5 gm/100 gm twice daily and cocaine hydrochloride at 20 mg/100 gm daily. In all groups the treatment started from the 6th day to the 15th day of gestation. The fourth group is the control. SETTINGS: Department of Anatomy, College of Medicine, King Faisal University, Dammam, Saudi Arabia and Department of Anatomy, College of Medicine, Al-Azhar University, Cairo, Egypt. INTERVENTION: The pregnant mice were sacrificed at the 20th day of gestation (one day before term). The uterine contents were evaluated and the number of dead and live foetuses was determined as well as the number of resorption. The living foetuses were examined for soft tissue anomalies. SUBJECTS: Eighty pregnant mice. RESULTS: This study revealed a significant increase in the number of dead foetuses and foetal growth parameters showed a significant decrease in the combined group. This group showed significant increase in anomalies of the head as hydrocephaly and complete cleft palate, of the kidney as bilateral secondary hydronephrosis and ectopic kidney, of the limbs, and of the tail as kinked tail. Both alcohol and cocaine groups alone showed significant increase in some of the anomalies of the head while in the alcohol group there was unilateral hydronephrosis. CONCLUSION: This study indicates that the combined administration of alcohol and cocaine has a more teratogenic effect than either alcohol or cocaine alone. PMID- 16261923 TI - Daytime encopresis associated with gland mal epileptic seizures: case report. AB - Sphincteric incontinence of stool and urine are not unusual features of generalised epileptic seizures. Isolated secondary encopresis as a manifestation of an epileptic seizure is unusual. This report is of, a four year old boy, with daytime secondary non-retentive encopresis. The onset of encopresis was preceded by several episodes of nocturnal generalised tonic clonic epileptic seizures. An electroencephalogram showed features consistent with complex partial seizures. He was commenced on anti-epileptic treatment with phenytoin sodium, and by the third day of treatment, the patient had achieved stool control. PMID- 16261924 TI - Mortality in over 350,000 insured Swedish dogs from 1995-2000: I. Breed-, gender , age- and cause-specific rates. AB - This study presents data on over 350,000 insured Swedish dogs up to 10 years of age contributing to over one million dog-years at risk (DYAR) during 1995-2000. A total of 43,172 dogs died or were euthanised and of these 72% had a claim with a diagnosis for the cause of death. The overall total mortality was 393 deaths per 10,000 DYAR. Mortality rates are calculated for the 10 most common breeds, 10 breeds with high mortality and a group including all other breeds, crudely and for general causes of death. Proportional mortality is presented for several classifications. Five general causes accounted for 62% of the deaths with a diagnosis (i.e. tumour (18%), trauma (17%), locomotor (13%), heart (8%) and neurological (6%)). Mortality rates for the five most common diagnoses within the general causes of death are presented. These detailed statistics on mortality can be used in breed-specific strategies as well as for general health promotion programs. Further details on survival and relative risk by breed and age are presented in the companion paper (Egenvall et al. 2005). PMID- 16261925 TI - Mortality in over 350,000 insured Swedish dogs from 1995-2000: II. Breed-specific age and survival patterns and relative risk for causes of death. AB - This study continues analysis from a companion paper on over 350,000 insured Swedish dogs up to 10 years of age contributing to more than one million dog years at risk during 1995-2000. The age patterns for total and diagnostic mortality and for general causes of death (trauma, tumour, locomotor, heart and neurological) are presented for numerous breeds. Survival estimates at five, eight and 10 years of age are calculated. Survival to 10 years of age was 75% or more in Labrador and golden retrievers, miniature and toy poodles and miniature dachshunds and lowest in Irish wolfhounds (91% dead by 10 years). Multivariable analysis was used to estimate the relative risk for general and more specific causes of death between breeds accounting for gender and age effects, including two-way interactions. Older females had tumour as a designated cause of death more often than males in most breeds, but not in the Bernese mountain dog. Information presented in this and the companion paper inform our understanding of the population level burden of disease, and support decision-making at the population and individual level about health promotion efforts and treatment and prognosis of disease events. PMID- 16261926 TI - Infrared thermography and ultrasonography to indirectly monitor the influence of liner type and overmilking on teat tissue recovery. AB - Eight Danish Holstein cows were milked with a 1-mm thick specially designed soft liner on their right rear teat and a standard liner mounted under extra high tension on their left rear teat. Four of the animals were overmilked for 5 min. Rear teats were subjected to ultrasound examination on the first day and to infrared thermography on the second day. Teats were submersed in ethanol 20 min post-milking on the second day. Ultrasonography measurements showed that teat canal length increased by 30-41% during milking. Twenty minutes after milking, teats milked with modified standard liners still had elongated teat canals while teats milked with the soft liner were normalized. Overmilking tended to increase teat wall thickness. Approximately 80% of variability in teat canal length, from before teat preparation to after milking, could be explained by changes during teat preparation. Thermography indicated a general drop in teat temperature during teat preparation. Teat temperature increased during milking and continued to increase until the ethanol challenge induced a significant drop. Temperatures approached pre-challenge rather than pre-milking temperatures within 10 minutes after challenge. Teat temperatures were dependent on type of liner. Mid-teat temperatures post-challenge relative to pre-teat preparation were dependent on overmilking. Thermography and ultrasound were considered useful methods to indirectly and non invasively evaluate teat tissue integrity. PMID- 16261927 TI - The effect of transdermal delivery of fentanyl on activity in growing pigs. AB - Recently, decreased activity levels have been observed in pigs treated postoperatively with transdermal delivery of fentanyl (TD-fentanyl) after isoflurane anaesthesia. Whether the change in behaviour is related to opioid induced sedation or to insufficient pain relief remains to be investigated. This study was therefore undertaken to evaluate the effect of TD-fentanyl 50 microg h( 1) on the activity level with and without isoflurane anaesthesia. Eight pigs (25.4 +/- 5.2 kg) were submitted to a cross-over study and given two treatments; 1) fentanyl patch applied after 30 minutes of anaesthesia (treatment A/F) and 2) fentanyl patch without anaesthesia (treatment F). The pigs' behaviour was observed from a video recording instantaneously every 10 minutes for 24 h before treatments and up to 72 h after the patch attachment. Venous blood samples were taken 1, 6, 12, 24, 48 and 72 h after the patch application. The behaviour recordings showed that TD-fentanyl did not produce sedation in any pig. No differences were found between the two treatments in activity level, weight gain or serum fentanyl concentration. This concentration measured after 24 h was 0.27 +/- 0.11 ng ml(-1) and 0.47 +/- 0.40 ng ml(-1) in the A/F and F group, respectively. In conclusion, transdermal delivery of 50 microg h(-1) fentanyl did not cause inactivity in growing pigs. However, the large variations in serum fentanyl concentration indicate that drug absorption from transdermal patches is unpredictable and sometimes deficient. PMID- 16261928 TI - Detection of antibodies to Erysipelothrix in stray dogs in Japan. PMID- 16261929 TI - Regulation of the erm(C) gene in Staphylococci from reservoir with different usage of macrolides. PMID- 16261930 TI - Anaplasma phagocytophilum infection in north Norway. The first laboratory confirmed case. PMID- 16261931 TI - A combination of atrioventricular block and sinoatrial block in a horse. PMID- 16261932 TI - Comparison of lethal versus non-lethal sample sources for the detection of infectious salmon anemia virus (ISAV). AB - The emergence of infectious salmon anemia virus (ISAV) in Canada and the USA has led to the establishment of ISAV surveillance programs for cultured Atlantic salmon (Salmo salar L.) and wild fish species, including Atlantic salmon. Current testing procedures for ISAV consist of viral culture, reverse-transcription polymerase chain reaction (RT-PCR) and indirect fluorescent antibody testing (IFAT), and require lethal sampling. As the focus of this study, blood was evaluated as a possible non-lethal sample source for ISAV diagnostic screening by viral culture and RT-PCR. Tissue samples (consisting of kidney/spleen for viral culture or kidney only for RT-PCR), blood and, to a lesser extent, mucus were tested from Atlantic salmon survivors of laboratory ISAV infection trials and moribund fish from marine salmon grow-out facilities participating in a USDA sponsored surveillance program. The trial fish represented a potential carrier population, while the surveillance fish were composed of moribund individuals from ISA clinical sites. Sample sources and diagnostic techniques were compared. Blood compared well to tissue samples for viral culture and produced a greater number of positives than did kidney samples for ISAV detection by RT-PCR. RT-PCR using both kidney and blood samples was determined to be a more sensitive assay than viral isolation. Mucus did not perform well in either assay compared to the other sample sources. Blood appears to be a reliable non-lethal sample source for the detection of ISAV by viral culture and RT-PCR in both moribund and asymptomatic fish. PMID- 16261933 TI - Infectious hematopoietic necrosis virus: monophyletic origin of European isolates from North American genogroup M. AB - Infectious hematopoietic necrosis virus (IHNV) was first detected in Europe in 1987 in France and Italy, and later, in 1992, in Germany. The source of the virus and the route of introduction are unknown. The present study investigates the molecular epidemiology of IHNV outbreaks in Germany since its first introduction. The complete nucleotide sequences of the glycoprotein (G) and non-virion (NV) genes from 9 IHNV isolates from Germany have been determined, and this has allowed the identification of characteristic differences between these isolates. Phylogenetic analysis of partial G gene sequences (mid-G, 303 nucleotides) from North American IHNV isolates (Kurath et al. 2003) has revealed 3 major genogroups, designated U, M and L. Using this gene region with 2 different North American IHNV data sets, it was possible to group the European IHNV strains within the M genogroup, but not in any previously defined subgroup. Analysis of the full length G gene sequences indicated that an independent evolution of IHN viruses had occurred in Europe. IHN viruses in Europe seem to be of a monophyletic origin, again most closely related to North American isolates in the M genogroup. Analysis of the NV gene sequences also showed the European isolates to be monophyletic, but resolution of the 3 genogroups was poor with this gene region. As a result of comparative sequence analyses, several different genotypes have been identified circulating in Europe. PMID- 16261934 TI - Intra- and interlaboratory performance of antibiotic disk-diffusion susceptibility testing of bacterial control strains of relevance for monitoring aquaculture environments. AB - In the course of an international research project on hazard analysis of antimicrobial resistance in SE Asian aquaculture environments, 2 European Union and 3 SE Asian laboratories attempted to harmonize a procedure for antimicrobial agent susceptibility testing based on disk diffusion (DD). For this purpose, a selected panel of 10 bacterial control strains of relevance for monitoring warm water aquaculture environments was sent by the central laboratory to the other participating laboratories. In each laboratory, 10 independently replicated DD determinations of each control strain to 6 antibiotics were performed using Iso Sensitest Agar (ISA) according to a standard operating procedure (SOP); in total, this study thus yielded 300 data sets for all 5 laboratories. At the end of the study, strain authenticity of subcultures of the control strains used by the respective participating laboratories was verified by the central laboratory. Based on the arithmetic mean of 10 inhibition-zone diameter measurements and standard deviation (SD), intralaboratory SD variations ranged from 0 to 2 mm when 79% of the recorded data sets were considered. In 8% of the data sets, the SD value exceeded 4 mm, which in most cases could be attributed to the fact that the data points for a given strain-disk combination were not normally distributed in one of the laboratories. At the interlaboratory level, 81% of the SD values based on global averaging of 50 data points per strain-disk combination were situated in the 0 to 5 mm range. Comparison with a minimal data set from literature of DD testing performed with Mueller-Hinton (MH) medium indicated that the use of either ISA or MH medium in DD testing has a limited impact on the method's precision among different laboratories. In conclusion, the current study has provided a validated SOP to promote the coordination and harmonization of DD susceptibility methodologies for aquaculture-associated organisms at an international level. As one of the main action items for the future, new interpretive breakpoints should be specifically designed and validated for aquaculture drugs and organisms. PMID- 16261935 TI - Respiratory pathogenesis of amoebic gill disease (AGD) in experimentally infected Atlantic salmon Salmo salar. AB - The aim of this study was to investigate the respiratory responses of Atlantic salmon, Salmo salar, experimentally affected with amoebic gill disease (AGD). In Series I, arterial blood samples were taken over a 96 h period following amoebae addition to examine potential respiratory effects associated with initial exposure. No major significant treatment effects were found between fish exposed to amoebae and control (non-exposed) fish. Arterial pH (pHa) was seen to be significantly elevated at 48 h in AGD fish relative to the 0 h time point. To investigate the long-term respiratory effects associated with infection, fish were similarly exposed to amoebae and sampled over a 16 d period. As for Series I, caudal blood pH was significantly elevated by Day 2 (48 h) compared to the pre (Day 0)-time point, suggesting that initial exposure to amoebae and/or amoebae attachment may have induced an initial respiratory alkalosis via increased ventilation frequency and/or amplitude. From Day 7 onwards, and coinciding with a significant increase in the percentage of affected gill filaments, blood pH decreased significantly, possibly indicating the onset of the characteristic respiratory acidosis that has previously been described for experimentally AGD affected Atlantic salmon. Although fish in this study showed up to 90% AGD affected filaments, the corresponding respiratory results do not reflect a major acid-base disturbance. Therefore, the findings from the present study support the contention that, although AGD only affects the gill, AGD-associated mortality in Atlantic salmon may not be primarily associated with respiratory failure. PMID- 16261936 TI - Two genotypic groups of morphologically similar fish trypanosomes from the Okavango Delta, Botswana. AB - Blood smears and blood lysate samples from freshwater fishes captured in the Okavango Delta, Botswana, were examined to determine whether their trypanosomes were all Trypanosoma mukasai, a species of supposed broad host specificity and widespread existence across Africa. Trypanosomes and/or babesiosomes occurred in 20/32 blood smears, and morphometric analysis of trypanosomes from 13/32 smears showed features suggestive of T. mukasai, including nuclear indices consistently >1. In 16/32 blood lysate samples from which DNA was extracted, trypanosome DNA was detected in 12/16 by PCR (polymerase chain reaction), using trypanosome specific ssu rRNA gene primers. Two samples positive for trypanosomes in blood smears yielded no amplifiable trypanosome DNA, but 4 samples with no detectable infection in blood smears were positive for trypanosome DNA, suggesting an overall trypanosome prevalence rate of 17/32 (53%) among fishes and demonstrating the value of PCR in trypanosome recognition. Cloning and sequencing of the 12 amplified fragments revealed 2 genotypic groups among these fish trypanosomes. Group 1 trypanosomes were from cichlids and 3 families of catfishes, Group 2 from 2 types of catfishes. Sequence comparison showed that the consensus Group 1 sequence was most similar to that of Trypanosoma cobitis, representing European fish trypanosomes of the carassii type, while the consensus Group 2 sequence showed similarity with a trypanosome sequence from another African catfish, Clarias angolensis. It was concluded that the identification of T. mukasai remains a problem, but at least 2 genotypic groups of trypanosomes occur in Okavango Delta fishes, and catfishes in this region appear to contain both types. PMID- 16261937 TI - Severe, chronic proliferative kidney disease (PKD) induced in rainbow trout Oncorhynchus mykiss held at a constant 18 degrees C. AB - Proliferative kidney disease (PKD), caused by the myxozoan parasite Tetracapsuloides bryosalmonae, is well documented as a seasonal disease of rainbow trout Oncorhynchus mykiss. Water temperatures influence the course of the infection both within the fish and the invertebrate host, the recovery of fish from the disease being accelerated with decreasing water temperatures. During this study, groups of rainbow trout were held at a constant temperature (18 degrees C) for a sustained period of time following initial exposure to T. bryosalmonae. While the majority of these fish had recovered from the clinical disease after 9 mo, 10% remained infected, showing clinical signs of disease. A histological study revealed that the majority exhibited very high parasite loads and unusually severe symptoms of PKD. This demonstrates that while most rainbow trout can recover from PKD independent of water temperature, there exists a sub population that cannot. PMID- 16261938 TI - Taxonomy and molecular phylogeny of Myxobolus bilobus n. sp. (Myxozoa) parasitizing Notemigonus crysoleucas (Cyprinidae) in Algonquin Park, Ontario, Canada. AB - Myxobolus bilobus n. sp. (Myxozoa) is described from golden shiner Notemigonus crysoleucas in Algonquin Park, Ontario, Canada. Plasmodia develop at the distal end of gill filaments, where they form a novel-shaped plasmodium made up of 2 side-by-side hemispheres joined at a central pore. Surrounding gill tissue is vacuolated, necrotic, and hemorrhagic. Spores of M. bilobus n. sp. resemble those of M. aureatus Ward, 1919, M. angustus Kudo, 1934, M. spalli Landsberg & Lom, 1991, and M. pseudokoi Li & Desser, 1985 in parasitizing cyprinids and in their pyriform shape. Spores of M. bilobus n. sp., however, are much larger (20 to 22.1 microm long, 7.5 to 9.3 microm wide, and 6 microm thick) than those of these other species, and the plasmodium is bi-lobed rather than the typical hollow sphere. A phylogenetic analysis of the 18S rDNA (2014 bp) reveals that M. bilobus n. sp. is a member of a clade that includes 11 species of Myxobolus, all of which are parasites of cyprinid fishes in North America or Eurasia. The study concludes that M. bilobus n. sp. is a member of a clade that has undergone radiation within cyprinid fishes of the 2 continents and that this evolution has involved both host and site switching. PMID- 16261939 TI - Life cycle studies of Myxobolus parviformis sp. n. (Myxozoa: Myxobolidae) from bream. AB - We experimentally followed the life cycle of Myxobolus parviformis sp. n., a myxozoan parasite from the gills of common bream Abramis brama L. Establishing the development of both stages (myxospore and actinospore) in 2 consecutive, full transmission trials, we were able to separate plasmodia of a common genotype from sympatric Myxobolus spp. occurring in naturally infected gill lamellae. Therefore, isolated gill plasmodia representing individual myxosporean 'clones' were used for subsequent infection of oligochaetes after molecular and morphological identification. The plasmodia of this species are located in median to distal regions of the primary gill filaments, forming small spherical pseudocysts. The comparatively small myxospores share some uniform characteristics: they taper posteriorly, have 2 inward inclining polar capsules that occupy half of the spores' interior space, and usually show 4 posterior sutural edge markings. The corresponding actinosporean has already been described morphologically and molecularly. The 18S rDNA sequence of the actinosporean stage was identical in all our samples, including myxospores. The triactinomyxon had a stout style, 32 sporozoites and short tapering caudal processes, and was shed by the tubificid oligochaete Limnodrilus hoffmeisteri. The ellipsoid sporoplasm was covered by a soft sheath which was emitted after valve shell opening upon stimulation by agitation or fish mucus. The molecular data (unique restriction fragment length polymorphism pattern and a 1586 bp 18S rDNA sequence) clearly differ from those for similar species and, together with the morphological data, justify the description of this parasite as a new species. PMID- 16261940 TI - Histopathology, ultrastructure and immunohistochemistry of Coregonus lavaretus hearts naturally infected with Ichthyocotylurus erraticus (Trematoda). AB - Histopathological, ultrastructural and immunohistochemical investigations were conducted on 26 specimens of powan Coregonus lavaretus (L.) from Loch Lomond (Scotland). The hearts of all 26 powan (15 females and 11 males) investigated harboured metacercariae of the digenean trematode Ichthyocotylurus erraticus (Rudolphi, 1809). The vast majority of metacercariae were located either singly or as an aggregation of white cysts on the surface of the bulbus arteriosus. The intensity of infection ranged from 2 to 200 larvae heart(-1), although the number of metacercariae found on male powan did not exceed 13. Histochemically, the parasite cyst wall gave a strong positive reaction with periodic acid schiff (PAS) and a faint positive signal with Azan-Mallory stain. All the metacercariae cysts were embedded in a granulomatous proliferation of heart epicardium tissue, forming a reactive fibroconnective capsule around the parasite. The capsule enclosing the parasite (produced by the host's reaction to the parasite) measured 13.57 to 90.20 microm (37.43 +/- 3.56) in thickness. Within the capsule wall, eosinophilic granular cells (EGCs), granulocytes, melanocytes and, in some instances, partially degenerated or vacuolated epithelioid cells were observed in close proximity to the cyst wall. Pigment-bearing macrophages were scattered throughout the granulomatous host-tissue reaction and as macrophage aggregates (MAs) within the capsules surrounding parasites. Immunohistochemical tests were applied to infected heart sections using 12 different antisera. Nerve fibres immunoreactive to bombesin, substance P (SP), and atrial natriuretic peptide (ANP) antisera were observed in close proximity to the parasite larvae. The presence of a serotonin-like substance was also observed within host immune-cells surrounding trematode cysts. Large cells of the epicardium were found to be immunoreactive to met-enkephalin and vasoactive intestinal peptide (VIP) antisera but not immunoreactive to anti-protein gene-product 9.5 (PGP9.5) sera. PMID- 16261941 TI - Use of molecular markers for species identification of Korean Perkinsus sp. isolated from Manila clams Ruditapes philippinarum. AB - Perkinsus is the pathogen responsible for mass mortality of the Manila clam Ruditapes philippinarum. Perkinsus sp. isolated from Manila clams collected in Korean waters was assayed by polymerase chain reaction (PCR) to determine its phylogenetic affinity with other congeneric species. Regions of rRNA of Perkinsus sp. isolated from clam haemolymph were cloned and sequenced. Sequences of a non transcribed spacer (NTS), internal transcribed spacers (ITS 1, 2) and 5.8S rRNA genes were compared to those available from other Perkinsus species. The NTS sequence of Korean Perkinsus was approximately 99.9 to 100% similar to that of P. atlanticus and 98.06 to 98.15% and 73.05 to 73.14% similar to those of P. olseni and P. marinus, respectively. The ITS 1, 5.8S rRNA and ITS 2 sequences of Korean Perkinsus showed 100% similarity to P. atlanticus and Perkinsus sp. reported from Japan. The ITS-5.8S rRNA sequences of Korean Perkinsus were 99.86 and 93.73% similar to those of P. olseni and P. marinus, respectively. The sporulation pattern and morphology of the Korean Perkinsus were very similar to those of P. atlanticus. Our data suggest that the Perkinsus sp. isolated from clams in Korean waters is P. atlanticus, which is currently synonymous with P. olseni reported from Australia. By considering that P. olseni has taxonomic priority, Korean Perkinsus sp. is accepted as P. olseni (atlanticus). PMID- 16261942 TI - Fenneropenaeus indicus is protected from white spot disease by oral administration of inactivated white spot syndrome virus. AB - Fenneropenaeus indicus could be protected from white spot disease (WSD) caused by white spot syndrome virus (WSSV) using a formalin-inactivated viral preparation (IVP) derived from WSSV-infected shrimp tissue. The lowest test quantity of lyophilized IVP coated onto feed at 0.025 g(-1) (dry weight) and administered at a rate of 0.035 g feed g(-1) body weight d(-1) for 7 consecutive days was sufficient to provide protection from WSD for a short period (10 d after cessation of IVP administration). Shrimp that survived challenges on the 5th and 10th days after cessation of IVP administration survived repeated challenges although they were sometimes positive for the presence of WSSV by a polymerase chain reaction (PCR) assay specific for WSSV. These results suggest that F. indicus can be protected from WSD by simple oral administration of IVP. PMID- 16261943 TI - [Controlled-release hydromorphone in elderly patients with severe pain of different etiologies. Results of an observational study]. AB - BACKGROUND: Elderly patients (older than 65) do not always receive adequate analgesic treatment, or else they suffer from side effects of the administered opioid. An alternative is oral controlled-release hydromorphone, the efficacy and tolerability of which in patientswith cancer-related pain and pain of other genesis has been confirmed in clinical studies. AIMS AND METHODS: A total of 2650 patients (effective group 2412 patients) average age 64.3 +/- 13.4 most of whom suffering from tumor-related (56.2%) or musculoskeletal (49.4%) pain were recruited to a multicenter observational study to investigate controlled-release hydromorphone administered mainly on an ambulatory basis. Pain intensity and quality of life were self-assessed by the patients and recorded in a questionnaire. RESULTS: Pain intensity decreased by 64.3% from 7.0 initially to 2.5 at the final examination (0 = no pain, 10 = most severe pain). Quality of life improved by 51.9%. This corresponded to a self-assessed decrease of impairment of quality of life from 45.9 initially to 22.1 at the final examination (0 = no impairment, 70 = most severe impairment). Opioid-type side effects, documented before initiating treatment, decreased appreciably under treatment with hydromorphone. The efficacy and tolerability of the medication, as well as patient compliance, were assessed by the participating physicians as "very good" or "good". CONCLUSIONS: Controlled-release, orally administered hydromorphone is well suited for the treatment of elderly patients with severe pain of different etiologies, in particular those with cancer pain. It is both effective and well tolerated, and has an appreciable positive impact on the patient's quality of life. PMID- 16261944 TI - [Observational study of blood pressure control and microalbuminuria in type 2 diabetics on irbesartan or irbesartan/HCTZ]. AB - BACKGROUND: Patients with type 2 diabetes and microalbuminuria have an elevated risk of developing diabetic nephropathy or cardiovascular complications. The effectiveness of AT1 blockers, in particular irbesartan, in these patients has been confirmed in controlled clinical studies. AIM, METHOD: An observational study involving 500 general practitioners and 9057 patients was carried out to establish which therapeutic aims can be achieved under realistic day-to-day conditions. Inclusion criteria were type 2 diabetes, hypertension > 140/90 mmHg and current treatment with irbesartan or irbesartan/hydrochlorothiazide (HTCZ). The observational study covered a period of 6 months. RESULTS: A blood pressure measurement < or = 130/80 mmHgwas considered optimal, and the target blood pressure was defined as < or = 140/90 mmHg. At the time of the baseline examination, the systolic pressure was outside the target in 80% of the patients, and outside the optimal range in 92%. At the second control measurement the respective figures were 37% and 69%. The diastolic blood pressure was outside the target pressure in 43 % (initially) and in 7% (second measurement) of the patients, and outside the optimal range in 77% (initially) and 43% (second measurement). At the start of the study, microalbuminuria was exhibited by 66.5% of the patients and after 6 months in 50.9%. CONCLUSION: The observational study confirmed the good effect of irbesartan and irbesartan/HCTZ, and the benefit of effective medication and intensive standardized care. PMID- 16261945 TI - [Efficacy of a combined Sabal-urtica preparation in the symptomatic treatment of benign prostatic hyperplasia. Results of a placebo-controlled double-blind study]. AB - This re-evaluation of a double-blind placebo-controlled therapeutic study of the combined sabal-urtica preparation PRO 160/120 investigates the changes in the irritative symptoms of benign prostatic hyperplasia (BPH) under the test substance in comparison with placebo. It was found that, over the study period of 24 weeks, the micturition symptoms frequency and urgency were statistically significantly improved under the well-tolerated PRO 160/120 in comparison with placebo. The patient's quality of life was also significantly better under PRO 160/120 in comparison with placebo. CONCLUSION: The often distressing symptoms of BPH can be effectively ameliorated already after only a few weeks of treatment with the sabal-urtica preparation PRO 160/120. In particular those patients with the stigmatizing symptoms urinary urgency and frequency benefit from such treatment. PMID- 16261946 TI - [Local tolerability of two preparations with eucalyptus oil and pine-needle oil]. AB - The objective of this open-label, single-centre study was to assess the tolerability of two locally applicable eucalyptus oil and pine-needle oil preparations using patch testing as an accepted and standardised method. METHOD: Two eucalyptus oil and pine-needle oil preparations were applied to the forearm of 46 subjects with healthy skin using standardised patches. The patches were removed after 48 hours and skin reactions were assessed immediately and after 24 and 48 hours. The major objective was the reaction of the patch test. RESULT: Neither preparation caused any positive skin reaction in any of the subjects. CONCLUSION: Skin tolerability to the two eucalyptus oil and pine-needle oil preparations is very good with regard to their active ingredients and other excipients. PMID- 16261947 TI - [Investigation of the effect of oxymetazoline on the duration of rhinitis. results of a placebo-controlled double-blind study in patients with acute rhinitis]. AB - AIMS AND METHOD: A prospective placebo-controlled double-blind study investigated the question whether oxymetazoline has a clinically relevant impact on the duration of acute rhinitis. The study included 247 patients with acute rhinitis who, over a maximum of 10 days, received either oxymetazoline hydrochloride (test substance group) or physiological saline solution applied in the form of a spray (control group). The main target measure was the duration of rhinitis, with time to onset of effect and symptoms as secondary measures. The safety of the treatment was determined on the basis of patient satisfaction, heart rate measurement, and undesired side effects recorded. RESULTS: The duration of the rhinitis decreased significantly under oxymetazoline in comparison with the control group (4 vs. 6 days). On average, the effect of oxymetazoline set in after 25 seconds, as compared with 90 seconds for physiological saline (difference significant, P < 0.001). From the second day of treatment, the symptoms improved significantly more markedly in the oxymetazoline group, and were identical again in both arms of the study from the 9th day onward. The physicians assessed the efficacy of oxymetazoline to be good orvery good in 85% of the cases as compared with 38% for the physiological saline solution (P <0.001). 84% of the patients in the oxymetazoline group assessed the effect to be good or very good, as compared with 44% in the patients in the physiological saline group (P < 0.001). CONCLUSION: All the parameters investigated revealed oxymetazoline to be significantly superior to physiological saline solution. Treatment with the former significantly shortened the duration of the rhinitis by one-third (2 days). PMID- 16261948 TI - [Systematic management of chronic wounds employing the TIME concept]. AB - The notion that chronic wounds are merely a variant of acute wounds is obsolete. The pathophysiology of chronic wounds differs in essence from that of acute wounds. The former is a specific pathological entity that requires both a systematic disease-specific diagnostic work-up and treatment. For the diagnosis and treatment of chronic wounds, the Wound Bed Preparation Advisory Board has proposed a new conceptthat goes bythe acronym of TIME, the letters of which refer to the structures to be diagnosed and treated: T = tissue, I = inflammation or infection, M = moisture (wound exudate), E = edge (of the wound). The present article discusses the principles and scientific background to this TIME concept. PMID- 16261950 TI - Cancer incidence in 13811 patients skin tested for allergy. AB - AIM: Several studies have shown a negative correlation between cancer and atopy related diseases. There are also a few reports of a positive relationship. We wanted to further evaluate these relationships in a prospective study. SUBJECTS AND METHODS: The incidence of malignant diseases among adult patients with atopy related diseases (asthma, rhinitis, urticaria, eczema etc; n = 13811), who had been skin prick tested in 1976-1999 was compared with the incidence in the general population. Expected cancer incidence from the date of skin prick testing up to 1999 was obtained from cause-, sex-, calendar-year-, and 5-year-age-group specific incidence rates for the county. These rates were calculated from cancer incidence and population counts obtained from the Swedish Cancer Register. The 95% confidence intervals (CIs) for cause-specific standardized incidence ratios (SIRs) were calculated. Skin prick tests were performed with Dermatophagoides pteronyssinus, horse, dog, cat, timothy, mugwort, birch, and Cladosporium. Patients having one or several positive skin prick test reactions (> or = 2+) were regarded as atopics. RESULTS: 119 cases of cancer occurred among 6224 atopic individuals (SIR 1.0) compared with 216 cases (SIR 0.94, CI 0.82-1.08) among 6358 non-atopics. There was a slight excess of Hodgkin's lymphoma cases among atopic men (SIR 4.03, 95% CI 1-10.3), and of non Hodgkin lymphoma cases among atopic women (SIR 4.52, 95% CI 1.23-11.6). However, a large number of comparisons were made which can have caused random findings. CONCLUSIONS: The results showed no associations between atopy or allergic symptoms, and subsequent cancer risk, but supported the theory that type-I allergy is not related to cancer risk. PMID- 16261949 TI - [The situation of patients with dementia may be rectified by Ginkgo biloba. Results of a health services research study concerning the ability of patients with dementia, quality of life of the nursing family members and total treatment costs]. AB - BACKGROUND AND ISSSUES: Ginkgo biloba-extracts are often used in therapy of patients with dementia. In this study, benefit and structure of Ginkgo biloba extract EGb 761 in treatment of patients with dementia was examined. PATIENTS AND METHODS: For the assessment of quality of life of care-taking relatives and patients as well as treatment costs were documented. The study was conducted as a non-randomised, two-armed cohort study with an open design for 683 slightly or moderately demented patients, aged between 65 and 80 years. Society's perspective was taken. Barthel-Index and MMST were also documented. Because of significant differences at inclusion of both cohorts, a matched-pairs-analysis and multiple regression analysis conducted. RESULTS: According to PLC a significant improvement in quality-of-life of care-taking relatives (p < 0.001) and patients (positive mood p = 0.018, negative mood p < 0.001) was only observed in the Ginkgo-cohort. Also Barthel-Index indicated an improvement in the Ginkgo-cohort (p < or = 0,001). MMST-scores increased significantly only in the Ginkgo-cohort (p < 0.001). Average total cost per patient amounted to 3.614,75 euro in the standard-cohort, whereas these costs per patient in the Ginkgo-cohort amounted to 3.031,78 euro (p = 0.067). Results were confirmed by matched-pairs-analysis. RESULTS: Ginkgo treatment has a valid place in caretaking structure of health services. Gingko attributes to a higher quality of life for both care-takers and patients, the progression of disease is slowed down and treatment costs are lower. PMID- 16261951 TI - Direct and prolonged exposure to dogs does not influence the degree of skin prick test positivity to dog allergen. AB - BACKGROUND: The relationship between pet ownership and the risk of developing allergic sensitization to pet allergens is still controversial. We assessed the possible effect of direct exposure to dog allergen on skin reactivity in dog sensitized patients. METHODS: We studied, in a case-control trial, 116 adults sensitized to dog allergens (55 with a dog at home for at least 10 years and 61 without it). The degree of response was assessed by skin prick test, performed in quadruplicate with three concentrations of allergenic extract: A (1:20 w/v), B (1:200 w/v) and C (1:2000 w/v). The mean diameter of each wheal was assessed using a visilog image analysis software. RESULTS: No significant difference between the two groups in the wheal diameters induced by the three concentrations of dog allergen could be demonstrated. CONCLUSION: The results of this study suggest that direct dog exposure in adults with respiratory allergy is not associated with greater cutaneous response to dog allergens, as compared to non exposed subjects. PMID- 16261952 TI - Changes in thymus- and activation-regulated chemokine (TARC) associated with allergen immunotherapy in patients with perennial allergic rhinitis. AB - BACKGROUND: Thymus- and activation-regulated chemokine (TARC), which is a CC chemokine receptor (CCR) 4 ligand with ability to recruit Th2 cells to inflammatory sites, is pathogenetically important in allergic rhinitis. Specific immunotherapy (IT), among the most effective therapies for allergic rhinitis, has incompletely understood mechanisms of action. TARC might be involved in some benefits of IT. METHODS: TARC in sera was assayed, obtained from 50 patients with house dust mite allergic rhinitis before and 1 year after beginning IT. Their ages ranged from 6 to 34 years (mean, 10), 30 were male and 20 were female. RESULTS: In patients whose nasal obstruction responded to IT, TARC decreased significantly with IT, while when response was defined in terms of sneezing or rhinorrhea, TARC did not change significantly. CONCLUSION: TARC might be an important target of IT in reducing obstructive allergic rhinitis. PMID- 16261953 TI - Distribution of primary immunodeficiency disorders diagnosed in the Children's Medical Center in Iran. AB - Primary immunodeficiency disorders include a variety of diseases that render patients more susceptible to infections. To determine the percentage of different primary immunodeficiency disorders diagnosed in the Children's Medical Center Hospital affiliated to Tehran University of Medical Sciences in Iran, we retrospectively reviewed the charts of the patients being referred to our hospital for immunologic evaluation of recurrent infections during a 20 year period. Among these patients, antibody deficiencies were the most frequent ones and were found in 52.6% of patients (n = 130). T-cell disorders, phagocytic disorders and complement deficiencies were found to be present in 24.69% (n = 61). 22.2% (n = 55) and 0.4% (n = 1) respectively. On the whole, common variable immunodeficiency was the most frequent disorder (n = 65), followed by ataxia telangiectasia (n = 39), X-linked agammaglobulinemia (n = 33), chronic granulomatous disease (n = 29) and selective IgA deficiency (n = 20). This study reveals that antibody deficiencies are the most common type of disorders as shown in other studies. A comparative study shows some differences between our results and other registries. This article also indicates that immunodeficiency disorders should be considered in patients with recurrent infections. PMID- 16261954 TI - Increase in prevalence of rhinoconjunctivitis but not asthma and atopic eczema in teenagers. AB - OBJECTIVE: We sought to assess changes in the prevalence of asthma and other allergic diseases in schoolchildren of Curitiba between 1995 and 2001. METHODS: Schoolchildren (aged 13-14) were evaluated by a self-reported written questionnaire of ISAAC protocol for symptoms of asthma and allergic diseases. The diagnoses of asthma, rhinoconjunctivitis and atopic eczema were considered according to the ISAAC protocol. Probable asthma was considered: more than 4 attacks of wheezing in the last 12 months; 1 to 3 attacks of wheezing in the last 12 months with dry cough at night, and/or with sleep disturbance due to wheezing, and/or with wheezing after exercise. The data obtained in 2001 were compared with the data obtained in 1995 for the same age group. RESULTS: We evaluated 2,946 schoolchildren in 1995 and 3.628 in 2001. The prevalence of symptoms of asthma, probable asthma, rhinoconjunctivitis and eczema in 1995 and 2001 was 18.4% and 18.7%, 11.6% and 12.4%, 14.2% and 17.2%, 3.8% and 3.7%, respectively. There was a 23.7% increase in the prevalence of rhinoconjunctivitis symptoms (p < 0.05). CONCLUSIONS: The prevalence of symptoms of asthma, probable asthma and eczema did not change over the past 6 years but a significant increase of the rhinoconjunctivitis symptoms was detected. PMID- 16261955 TI - Mite allergen exposure, sensitisation and clinical symptoms in Valdivia, Chile. AB - One hundred consecutive asthmatic paediatric patients were evaluated and skin tested with a battery of skin prick test reagents, including 8 different standardized house dust mite extracts. Asthma severity was graded according to the Global Initiative for Asthma (GINA) document in mild persistent (52 patients), moderate persistent (39) and severe persistent (9). Sixty patients had asthma and allergic rhinitis, 12 asthma and eczema, and 8 asthma, allergic rhinitis and eczema. The patient population was divided into 2 different socioeconomic groups (50 patients per group) based on a standardized, validated questionnaire. A dust sample was collected with an adapted vacuum cleaner from the mattress of each patient and analysed for Der p 1, Der f 1 and Der p 2 allergen content using monoclonal antibodies. Eighty patients were skin test positive to at least one mite species. All positive skin test patients were positive to Dermatophagoides pteronyssinus, 99% to D. farinae, 92% to Euroglyphus maynei, 80% to Lepidoglyphus destructor, 73% to Tyrophagus putrescientae, 72% to Blomia tropicalis; 70% to Acarus siro and 68% to Chortoglyphus arcuatus. All patients with severe persistent asthma had a positive skin test to mites, 85% in the moderate group, and 73% in the mild group (p < 0.01). 95% of patients with asthma and allergic rhinitis had a positive skin test to mites, 92% of patients with asthma and eczema and 100% of patients with asthma, allergic rhinitis and eczema; (p < 0.01). Mean Der p 1, Der f 1 and Der p 2 allergen concentrations were 18.3, 0.6 and 5.6 microg/g of mattress dust, respectively. Mean Der p 1 allergen levels in the middle-low socioeconomic group were significantly higher than in the middle high group (p < 0.01). There is a high rate of allergic sensitisation among pediatric asthmatic patients in Chile. More than one species are implicated, although sensitisation and exposure to D. pteronyssinus predominates. Mite allergic patients are exposed to high mite allergen concentrations, exceeding previously established risk levels for sensitisation and symptoms. PMID- 16261956 TI - Acute effect of nebulized budesonide in asthmatic children. AB - The acute anti-inflammatory effects of inhaled steroids at high doses and their use at home and as emergency treatment of acute asthma attacks in children have been evaluated in many clinical studies. However very little is known about their additional bronchodilator response to systemic steroids plus nebulized salbutamol in the early management in children. Asthmatic patients aged between 5-15 years were investigated in a double-blind, placebo-controlled fashion. Both the study group (Group I) and the control group (Group II) received three consecutive doses of nebulized salbutamol (0.15 mg/kg/dose) and one dose of parenteral methylprednisolone (1 mg/kg/dose, intramuscularly). After this treatment, nebulized budesonide (1 mg/dose) was administered to patients in the study group and placebo (nebulized saline) was administered to patients in the control group. Pulmonary index scoring and peak flow meter was performed to both groups before and after the treatment. There were twelve patients in Group I (mean age: 7.90 +/ 2.34 years) and fourteen patients in Group II (mean age: 9.36 +/- 2.55 years). There was no difference between the two groups with respect to age (p = 0.1421), gender (p = 1.000) and inhaled steroid prophylaxis rate (p = 0.2177). No statistically significant difference was detected between the two groups with respect to the pulmonary index score (p = 0.3528). Yet, there was a statistically significant difference between the two groups with respect to the increase in PEFR (p = 0.0155). The positive acute effect of nebulized budesonide in addition to systemic steroids and nebulized salbutamol in improving the spirometric indices in asthmatic children is an encouraging finding for further investigations of its routine use in the pediatric emergency department. PMID- 16261957 TI - Validation of the Spanish version of the Phase III ISAAC questionnaire on asthma. AB - The prevalence of allergic diseases in childhood has increased significantly over the last decades. This increase seems to be closely associated with the way of life of western societies. The high prevalence differences on different regions may be due to linguistic and cultural reasons and not to real variations in prevalence. This is the reason why several authors felt the need to perform an objective validation of their versions. Our working group has published the results of the Phase I validation and now is publishing the Phase III validation in order to guarantee the reliability of this phase results. The study sample is formed by 366 children aged 3 to 17 years. The following steps were followed in this study: I. Assessment of the "Criterion validity" of the Spanish ISAAC Bronchial Asthma questionnaire, evaluating the sensitivity, specificity, relative value, and positive and negative predictive values. 2. Determine the questionnaire reliability, analysing its "Inner consistency". 3. Statistical comparison between our ISAAC-Bronchial Asthma results and the ones obtained by other groups (external concordance and consistency), in order to prove the previously evaluated reliability. 4. Comparison between the ISAAC-Bronchial asthma questionnaire diagnostic ability and the standard diagnostic criteria universally used in clinical praxis. We could confirm that there is a high and very significant concordance between the questions aimed to detect children with asthma. In this sense, it is especially useful the question about "ever had wheezing" because of its high sensitivity (93.3%) and specificity (89.9%), that make it able to be used as initial screening test in a general population, and that has shown a high concordance percentage with the questions "ever had asthma" (98%), "wheezing with exercise" (75%), and "cough at night"(80%). The questions that give more information about the evolution and control of the asthmatic disease are "wheezing in the last 12 months", "number of attacks in the last 12 months", "wakening at night", "wheezing with exercise" and "dry cough at night in the last 12 months". The questions more related to asthma severity were "number of attacks in the last 12 months", "wakening at night", "stop speaking in order to breath", and "wheezing with exercise". We conclude that ISAAC-Asthma questionnaire Phase III is a useful tool for the assessment of childhood asthma due to its criterion validity, inner consistency and external concordance. PMID- 16261958 TI - Hypoxemia: an early indication of pigeon breeders' disease. Clinical and laboratory findings among pigeon breeders in the Salonica area. AB - The purpose of this study is to identify pathological findings among pigeon breeders in the Salonica area. Fifty-four volunteer breeders with varying intensity of contact with pigeons participated in the study. All the breeders, after filling in a questionnaire that included questions about the existence of conjunctivitis, rhinitis, chronic cough and sputum, were subjected to clinical examination and spirometric, hematological (arterial and venous), radiographic and immunologic tests. Twenty-five point nine percent of the breeders suffered from conjunctivitis, 31.5% from rhinitis and 33.3% from chronic cough and sputum. Fourteen point eight percent of them had class I precipitins, 7.4% had class II, 5.6% had class III and 16.7% had class IV and a positive correlation of precipitin class with the number of pigeons bred was found. Seven breeders (13%) had hypoxemia at rest and other 8 (14.8%) presented with hypoxemia after the exercise testing. Two breeders presented with a combination of findings of allergic alveolitis that satisfied the criteria of the Pigeon Breeders' Disease (PBD). A substantial number of the examined pigeon breeders were sensitized to pigeon serum antigens. Arterial hypoxemia, either at rest and/or after exercise was observed in 27.8% of breeders. Three point seven percent of breeders presented with clinical and laboratory findings consistent with PBD. Chronic cough and sputum in pigeon breeders should be considered as a possible manifestation of PBD. PMID- 16261959 TI - Allergy to sea fishing baits. AB - We report a new case of rhinitis and asthma caused by sea fishing baits. The results showed exposure to Sipunculus nudus (Phylum Sipuncula; order Sipunculida: Sipunculidae) to be the main cause of the allergic symptoms. The intervention of IgE was demonstrated, with the presence of cross-reactions with allergenic extracts from other worm species used as baits, belonging to different orders of Annelida. PMID- 16261960 TI - Occupational asthma caused by white mushroom. AB - The commercial growing of white mushroom (Agaricus bisporus) is a frequent activity in certain Spain regions as La Rioja. We report two cases of white mushroom workers suffering from asthma caused by hypersensitivity to basidiocarp and spores of white mushroom. PMID- 16261961 TI - Acute pancreatitis possibly caused by allergy to bananas. AB - We are reporting a forty-seven year old female who had three attacks of acute pancreatitis after having; eaten bananas. She was treated with a light diet and intravenous fluids during each of her admissions. The treatment concluded with the disappearance of clinical symptoms such as epigastralgia, nausea, vomiting and diarrhea within a few days. In addition, elevated serum and urine amylase levels returned to normal values in parallel with the clinical symptoms. The data during her three attacks of acute pancreatitis were as follows: serum total IgE level = 644 IU/mL, specific IgE to bananas = 2.18 UA/ml. No remarkable abnormalities were present in sonography, computed tomography, magnetic resonance imaging of the abdomen, and magnetic resonance cholangiopancreatography. Endoscopic examination of the upper digestive tract showed the ampulla of Vater swollen and edematous, thus a biopsy was performed. Toluidine blue staining and immunohistochemical staining against human mast cell tryptase with the biopsy specimens showed mast cells accumulating in mucosa and submucosa. By avoiding consumption of bananas, she has not suffered from any additional attacks of pancreatitis since the third attack in the last thirty-four months up to this day of December 10, 2004. PMID- 16261962 TI - Anaphylaxis due to Brazil nut skin testing in a walnut-allergic subject. AB - The diagnosis and management of nut allergy can be difficult because of the possible severity of the clinical manifestations and the cross reactivity between different species. We report a case of anaphylaxis due to skin testing in a young adult with clinically ascertained walnut allergy. After an episode of anaphylaxis due to walnut ingestion, a routine diagnostic workup was carried out, involving skin prick test with commercial extracts, prick by prick with fresh food and CAP RAST assay for different nuts. Immediately after pricking with fresh Brazil nut, a severe episode of anaphylaxis occurred, that required epinephrine and intravenous steroids. The subject had never eaten Brazil nut before. Therefore we hypothesize a cross reactivity effect, since this phenomenon is well known for tree nuts. Our case suggests that in vivo diagnosis, especially if fresh nuts are used, should be performed only if adequate equipment to treat anaphylaxis is available. PMID- 16261963 TI - Allergy to mammal's meat in adult life: immunologic and follow-up study. AB - Allergy to bovine meat and Bovine serum albumin (BSA) is exceptional, especially in the adult life. BSA is considered a minor allergen in cow's milk allergy, but there is little information about this antigen in reactions produced by other beef products as meat. To our knowledge, evolutive studies of beef's allergic patients have not been reported. OBJECTIVE: To present one patient with several allergic reactions (urticaria-angioedema) after eating different mammals' meat. METHODS: The patient underwent allergy testing through skin prick test (SPT), specific IgE detection and SDS-PAGE Immunoblotting and Immunodot inhibition studies. Periodic determinations of specific IgE to meats and epithelia were performed. RESULTS: Routine studies for chronic urticaria were normal or negative. SPT showed positive responses to pork, cow, rabbit and lamb meat, and dog, pork, sheep and cow epithelia. It was negative to cat, horse, guinea pig, rabbit, lamb, mouse epithelia, mixture of feathers, cow milk, soybean, mustard, mites and chicken meat and Anisakis simplex. Intradermal testing to BSA was positive. Determinations of specific IgE were positive to beef meat, lamb meat, pork meat and rabbit meat, dog, cat, cow, sheep and pork dander, cow's milk, and negative to chicken meat. Immunoblot and immunodot studies showed IgE recognition bands to bovine and lamb meat which were totally inhibited by BSA. A progressive reduction of the total and specific IgE, the latter until its total negativization, has been observed in the following three-year period. CONCLUSION: We report a case of IgE-mediated urticaria-angioedema due to BSA hypersensitivity, possibly induced by a subclinical sensitivity to dog and cat epithelium. The exclusion diet in patients allergic to these foods may be a progressive loss of clinical allergy. PMID- 16261964 TI - Realising potential. PMID- 16261965 TI - A night at the opera. PMID- 16261966 TI - Best boot forward. PMID- 16261967 TI - Investing in mental health. PMID- 16261968 TI - Constitutional racism is unwritten yet it governs the mindset of British citizens. PMID- 16261969 TI - My advice is to look for the positives--the negatives will soon disappear. PMID- 16261970 TI - Taking the first step forward. PMID- 16261971 TI - Knowledge, skills, values. PMID- 16261972 TI - We want to be able to work. PMID- 16261973 TI - Improving combat medic learning using a personal computer-based virtual training simulator. AB - The new military occupational specialty for combat medics, the 91W, requires that all medics successfully pass the National Registry of Emergency Medical Technicians examination. The objective of this study was to supplement standard emergency medical technician training with a three-dimensional, computer-based, virtual training simulator and to assess whether scores and pass rates could be increased. Combat medics (N=167) were selected for training through the usual noncommissioned officer unit selection process and were randomized by cohort to the training simulator. Results showed no significant differences in National Registry of Emergency Medical Technicians examination scores (t = 1.019, df = 153, p = 0.745, one-tailed) or pass rates (chi2 = 1.575, df = 1, p = 0.209). The findings, however, were used to construct two models of combat medic characteristics that can be used to assist in initial selection for emergency medical technician courses and subsequent counseling of soldiers on course completion. With further research, these models could be refined for Army-wide use to increase the cost-effectiveness of combat medic recruitment, training, and testing. PMID- 16261974 TI - Difference in fitness for duty among soldiers on a mission: can these be explained by a difference in the preemployment assessment? AB - In 1998, the Royal Netherlands Army introduced a new examination system (abbreviated as BMEKL), which was based on the "workload-capability" model, to replace the old system (abbreviated as PULHEEMS), which focused on diagnosis and was based solely on the detection of diseases and infirmities. To discern differences under operational conditions between soldiers examined with one of the two medical examination systems, we performed a prospective cohort study. In the study, soldiers who had been declared fit for duty with one of the two medical assessment systems (randomized) and sent on a mission were monitored for 2 years. We used the two operational measures of availability and health care costs. In addition, the candidates were given a questionnaire twice per year during the study period. The study revealed that the soldiers assessed using the function-based BMEKL system displayed greater fitness for duty than did those assessed using the diagnosis-based PULHEEMS system. The BMEKL assessment system is a better predictor of the ability to function as a soldier in general, and with regard to deployment, health, and the locomotor apparatus specifically, than is the PULHEEMS system. PMID- 16261976 TI - Congenital hypoplasia of the inferior vena cava: an underappreciated cause of deep venous thromboses among young adults. AB - This report illustrates how newer imaging techniques are identifying vasculopathies as risk factors for deep venous thrombosis (DVT). In this case, a healthy young man presented with a straightforward DVT but without traditional risk factors. Doppler ultrasonography confirmed a proximal clot, and contrast enhanced computed tomography identified a hypoplastic inferior vena cava (IVC). DVTs cause considerable morbidity and death each year, including approximately 200,000 fatal pulmonary embolisms. Specific treatment of DVTs and long-term management and prevention strategies are contingent on the etiology, which can be determined in approximately 85% of cases. A hypoplastic IVC was discovered during efforts to find the cause of thrombosis, and a Medline search suggests that this anomaly should be considered for young persons with DVTs. DVTs are common and their underlying etiology should be identified to help guide long-term management. Evidence is emerging that an anomalous IVC should be ruled out as a cause of DVT among young patients. PMID- 16261975 TI - Acute dysuria among female soldiers. AB - OBJECTIVE: To assess behavior patterns among active duty female soldiers presenting to military care facilities for acute dysuria. METHODS: A self administered questionnaire was developed. One hundred twelve female soldiers presenting with acute dysuria and one hundred twenty-six presenting for other reasons were surveyed. RESULTS: During regular duty hours, the dysuria group drank less than the control group (21% and 14%, respectively; p = 0.004). However, field duty appeared to compound this problem, with the dysuria group drinking considerably less than the control group (79% and 19%, respectively; p = 0.002). Both groups stated that they postponed urination during working hours. The dysuria group postponed urination more than did the control group during regular duty (75% and 53%, respectively; p = 0.006) and field duty (79% and 65%, respectively; p = 0.008). CONCLUSIONS: Female soldiers presenting with dysuria were more likely to report fluid restriction and to postpone voiding during duty, behaviors that were reported even more frequently during field duty. Fluid restriction and postponed urination may be significant factors in the development of acute dysuria among female soldiers. PMID- 16261977 TI - The impact of laparoscopic resection rectopexy in patients with total rectal prolapse. AB - Total rectal prolapse is a disabling disease. The aim of this study was to evaluate pain management, hospital stays, constipation, and continence status among military personnel who underwent laparoscopic surgery. Forty patients (mostly men) underwent laparoscopic rectopexy (LR) or laparoscopic resection rectopexy (LRR). Colonic transit time, postoperative pain scores, preoperative and postoperative anal function, and changes in constipation were assessed. The median operation times for LR and LRR were 126 and 223 minutes, respectively. The median postoperative hospital stays were 3 and approximately 6 days for LR and LRR, respectively. Patients needed fewer analgesics in a short postoperative period. However, there was no difference between the two groups in analgesic requirements. Continence improved for approximately 71% of patients, but constipation was treated for 50% of affected patients. No recurrences were noted in the follow-up periods, which were 13 and 22 months for the LRR and LR groups, respectively. The quality of life for the patients who underwent LR was not as good as that for the patients who underwent LRR, at the end of 1 year. We eliminated total rectal prolapse and almost cured incontinence by using laparoscopy, although the disadvantageous aspects were long operation times and suboptimal healing with respect to constipation and related symptoms. LRR is the more feasible procedure, with the emphasis on elimination of incontinence and constipation, producing a better quality of life for patients, in addition to short hospitalizations, necessity for analgesia for a short time, and return to hard training field activities in a short time among military personnel. PMID- 16261978 TI - Strategic planning for military occupational health care. AB - OBJECTIVE: This article describes a framework and system for planning appropriate occupational health contributions to military manpower availability and operational capability, and auditing the complete and effectiveness of implementation. METHODS: Prepared through author-led consensus-seeking serial deliberations with occupational health experts over initial and consequential drafts of, latterly, a populated strategic planning framework based on key stages in a military career. Illustrative examples are provided from the author's experience and, along with referenced sources of further information, the scientific literature. RESULTS: The resulting framework facilitates timely management of occupational health risks to recruiting, retention, operational availability and capability, and the general well-being of military personnel. Monitoring and audit are integral to the activity to provide an evidence-based spiral of refinement of the plan and improvement of outcomes. CONCLUSIONS: Key stages of the military career may provide manageable challenge bites for planning with executive and health care colleagues to forecast and control occupational health risks to operational capability and thus contribute to achieving military objectives. PMID- 16261979 TI - Risk of accidents and occupational diseases among the Finnish Defence Forces. AB - OBJECTIVES: Military activities are often considered more dangerous than civilian work, especially in crisis situations, but peacetime or even peacekeeping conditions have seldom been analyzed in this respect. According to the compensation statistics of the Finnish State Treasury, in 2001, 80% of injuries among military personnel were caused by accidents at work, 15% occurred during commuting to and from work, and 5% were attributable to occupational diseases. RESULTS: The compensated accident frequencies varied between 22 and 26 cases per 1 million work hours during the 1990s. During the past few years, the incidence rate has slightly declined. When accidents among military personnel were considered, most occurred during military exercises (57%), during other work (35%), or during work commuting (9%). One person died in an accident during a military exercise in 2001. On the basis of the data of this study, the hypothesis of a substantially higher risk during peacetime for military work, compared with civilian work, is not defensible. The accident rates in the calendar year 2000 were 25% higher for Finnish civilian work (32 cases per 1 million hours) than for the Finnish Defence Forces (22 cases per 1 million hours). PMID- 16261980 TI - Physical fitness after apical resection for the treatment of primary spontaneous pneumothorax. AB - The objective of the present study was to assess the physical fitness of patients after apical resection and partial apical pleurectomy for the treatment of primary spontaneous pneumothorax (PSP). Between 1982 and 1999, 58 patients received surgical treatment for PSP in our department. Twelve patients needed bilateral surgical intervention. At an average of 121 months after surgery (range, 16-231 months), the patients underwent follow-up assessments. Information was obtained on the basis of a questionnaire and from clinical examinations, including spirometry/body plethysmography and exercise testing with a bicycle ergometer. High-resolution computed tomography was used to identify postoperative changes of the lung apex. Forty-eight of 58 patients took part in the study, and all were found to be fully fit. High-resolution computed tomography gave evidence of new postoperative fibrocystic processes in 26 of the 31 affected apexes. One recurrence was observed (3.2%). Because apical resection cannot counteract pathogenetic mechanisms underlying parenchymal destruction and the formation of postoperative bullae among patients with a history of PSP, additional treatment of the apical pleura is necessary to prevent recurrences. Our results suggest that the physical fitness of patients with PSP can be completely restored postoperatively. PMID- 16261981 TI - Epidemiological review of insect sting allergy in naval aviation: current policy and real-world practices. AB - Individuals with conditions not addressed in the physical standards section of the Navy's Manual of the Medical Department can be considered for a waiver that would allow them to continue in naval aviation. Insect sting allergy is addressed in the Navy's waiver guide; however, the actual disposition of these individuals does not coincide with published waiver policies. Our objective was to identify discrepancies, to review current clinical guidelines, and to offer recommendations for updating the Navy's waiver policy. Aviation medical records of individuals with insect sting allergies from 1985 to 2002 were reviewed. Disposition, waiver status, and allergy evaluation were investigated. The data suggest that waiver requests have been addressed not under the current waiver guidelines but instead under current clinical guidelines. New guidelines that properly reflect current diagnostic and treatment modalities were voted on by the Aeromedical Advisory Council and submitted to the Navy's Bureau of Medicine. These changes were incorporated into the Navy's aviation waiver policy guidelines. PMID- 16261982 TI - Issues related to the use of tourniquets on the battlefield. AB - On the battlefield, a properly applied tourniquet can be an extremely effective means of controlling severe extremity wound hemorrhage. However, a great deal of confusion exists among soldiers, medics, and military medical officers on a number of tourniquet-related issues. What is an appropriate combat tourniquet? When is it appropriate to use a tourniquet? When and by whom should a tourniquet be removed? Under what conditions should a tourniquet not be released or removed? What are the most effective ways to increase limb salvage while using a tourniquet? These and other issues were addressed by a panel of experts at the 2003 Advanced Technology Applications for Combat Casualty Care Conference, August 21 and 23, 2003, St. Pete Beach, Florida. Here we review those issues and present a summary of the panel's recommendations. PMID- 16261983 TI - Physiological evaluation of the U.S. Army one-handed tourniquet. AB - OBJECTIVE: To provide a physiological assessment of the U.S. Army one-handed tourniquet (OHT). METHODS: An OHT was self-applied by 26 subjects, to maximal tolerable tightness, to the proximal arm or thigh under different conditions and positions, and the presence of blood flow was assessed using Doppler ultrasonography or occlusion plethysmography. RESULTS: Doppler sound was eliminated at the radial artery for all subjects with OHT application but was not stopped at the popliteal or dorsalis pedis artery for any subjects. The OHT reduced forearm blood flow by 79% but decreased leg blood flow by only approximately 50%, regardless of condition and position of application to the thigh. CONCLUSIONS: The OHT appears to effectively minimize blood flow in the arm but not in the lower extremities, and clinical assessment of blood flow disappearance by Doppler ultrasonography may underestimate the magnitude of actual blood flow to the limb. PMID- 16261984 TI - Exploring veteran identity and health services use among Native American veterans. AB - The objectives of this study were to describe the military experiences of Native American veterans and to explore how factors related to veteran identity influence their use of health services. Study participants completed a demographic and health questionnaire, followed by participation in a focus group session. The findings revealed that, despite their negative experiences during military service, most participants had a positive veteran identity. Almost 46% of participants reported having a service-related illness or injury. Almost one third (28.2%) used the Indian Health Service (IHS) exclusively for their health care, followed by those who used both IHS and Veterans Affairs (VA) services (23.5%), followed by VA-only users (21.2%). We conclude that Native American veterans highly identify with their military service but may turn to IHS for their medical care. The data support the current VA policy of strengthening coordination with the IHS to ensure that the medical needs of Native American veterans are addressed. PMID- 16261985 TI - Lifetime sexual and physical victimization among male veterans with combat related post-traumatic stress disorder. AB - Because of the high prevalence of post-traumatic stress disorder (PTSD) among veteran men and the limited research on victimization in this group, we recruited 133 male veterans with combat-related PTSD from a psychiatric inpatient unit and assessed them for lifetime physical and sexual trauma. Results indicated that 96% of the sample had experienced some form of victimization over their lifetimes; 60% reported childhood physical abuse, 41% childhood sexual abuse, 93% adulthood physical assault, and 20% adulthood sexual assault. In the preceding year alone, 46% experienced either physical or sexual assault. These findings support the need for routine inquiry into the histories of noncombat victimization in this cohort. Determining the lifetime history of trauma exposure may have implications for vulnerability to subsequent development of PTSD and the risk of future violence. PMID- 16261986 TI - Smoking and other factors influencing the oral health of Lithuanian Army recruits. AB - The aim of our research was to determine the prevalence of smoking among Lithuanian army recruits and how smoking and other factors affect oral health. The findings of our research showed that 70% of recruits smoke. The analysis of the research findings showed that smoking had a negative effect on oral hygiene. Especially smoking was harmful with respect to periodontal tissues. The periodontal lesions were more prevalent and severe among recruits who smoked. Smokers had a higher Community Periodontal Index of Treatment Needs index than nonsmokers. The analysis of the research findings showed that the state of oral health was related to other factors, such as oral hygiene, age, and education. It was determined that the oral hygiene of males living in the countryside and having poorer education was worse than that of males living in the city. Their status of periodontal tissues was worse, and they had more decayed and untreated teeth. The findings of the logistical regression analysis showed that poor education and living in the country, irregular tooth brushing, poor oral hygiene, and smoking were the most important factors related to a great number of untreated decayed tooth surfaces. PMID- 16261987 TI - Osteochondroma of the mandibular condyle: report of a case and review of the literature. AB - Osteochondromas rarely affect the mandibular condyle. An unusual case of an osteochondroma occurring in the left mandibular condyle in a 40-year-old man who presented with mandibular deviation and malocclusion is reported; this represents the 37th documented case in the English-language literature. The tumor was resected through condylectomy. Four-year follow-up assessments revealed satisfactory function and occlusion, without evidence of recurrence of the tumor. PMID- 16261988 TI - Determination of gunshot residues with image analysis: an experimental study. AB - In firearm injuries, assessment of the firing range and determination of entrance and exit wounds are important. For this reason, evaluation of the amount and distribution of gunshot residues (GSRs) is necessary. Several methods and techniques for GSR analysis have been developed. Although these methods are relatively sensitive and specific, they may require expensive dedicated equipment. Therefore, a simple, easily applicable, more convenient method is needed. A total of 40 experimental shots were made to calf skin from distances of 0, 2.5, 5, 10, 20, 30, 45, and 60 cm. Eighty samples were taken from the right and left sides of the wounds, and Alizarin Red S dye staining was performed. The amounts of GSR particles were measured with image analysis. GSRs were detected in all shots. The mean size of the distribution area of barium and lead elements around the wound had a significant negative correlation with increasing shooting distance (r = -0.97, p < 0.001). As the distance increased, the amount of GSR decreased, and this decrease rate was nonlinear. Variance analysis suggested significant differences between data groups depending on range (p < 0.001). The image analysis method may solve some of the standardization problems for evaluation of GSRs. GSR detection with the image analysis method does not require experienced personnel and may be a suitable method for scientific studies and for routine purposes. PMID- 16261989 TI - Myotonias and army personnel: symptoms and effects on service fitness. AB - Myotonias are rare disorders characterized by difficulties in skeletal muscle relaxation. Either dominant or recessive modes of inheritance are possible. Underlying gene mutations cause defects in the ion channels of the muscle membranes. Previously undiagnosed myotonias may occur among military conscripts. We report here eight such patients with enhanced symptoms of myotonia during their military service. Six patients had myotonia congenita, one had myotonic dystrophy, and one paramyotonia congenita. In myotonia congenita, serum creatine kinase and aldolase levels correlated with the recommended service fitness classification. Because some anesthetic agents may have unfavorable side effects in myotonia, both patients and anesthesiologists need to be aware of the diagnosis. The awareness of military surgeons regarding the possibility of myotonia is necessary to provide a correct diagnosis and to establish the service fitness of these patients. PMID- 16261990 TI - Delivery of medical services to different national groups during the war: an example from Bosnia and Herzegovina, 1991-2000. AB - OBJECTIVE: To investigate the delivery of health services to different ethnic groups in the war and postwar periods in the area of Livno, Bosnia and Herzegovina. METHODS: The total population, percentages by nationality, and numbers of persons medically or surgically treated at the War and General Hospital of Livno between January 1991 and October 2000 were retrospectively analyzed. Demographic data from church, military, and local government archives of the Livno region were used. RESULTS: In the prewar period (1991-1995), there had been living approximately 40,000 inhabitants (Croats, 75%; Moslems, 17%; Serbs, 8%) and 685,361 patients were treated (Croats, 46%; Moslems, 33%; Serbs, 21%). In the war period, 1590 patients were treated surgically (Croats, 78.5%; Moslems, 16.8%; Serbs, 4.7%). In the postwar period (1996-2000), surgery was performed for 3217 patients (Croats, 76.8%; Moslems, 19.3%; Serbs, 3.9%). CONCLUSION: The constant numbers of patients of different nationalities treated during and after the war reveal that the medical staff worked in an ethical and highly professional manner. PMID- 16261991 TI - Comparative effects of vitamin K and vitamin D supplementation on calcium balance in young rats fed normal or low calcium diets. AB - We examined the effect of vitamin K and vitamin D supplementation on calcium balance in young rats fed a normal or low calcium diet. Eighty female Sprague Dawley rats, 6 wk of age, were randomized by the stratified weight method into eight groups with 10 rats in each group: 0.5% (normal) or 0.1% (low) calcium diet, 0.5% or 0.1% calcium diet+vitamin K (vitamin K2, menatetrenone, 30 mg/100 g, food intake), 0.5% or 0.1% calcium diet+vitamin D (25 microg/100 g, food intake), and 0.5% or 0.1% calcium diet+vitamin K+vitamin D. The duration of the study was 10 wk. Vitamin K supplementation promoted the reduction in urinary calcium excretion and retarded the abnormal elevation of serum PTH level in rats fed a low calcium diet, and stimulated intestinal calcium absorption in rats fed a normal calcium diet. Vitamin D supplementation stimulated intestinal calcium absorption with prevention of the abnormal elevation of serum PTH levels and prevented hypocalcemia in rats fed a low calcium diet, and stimulated intestinal calcium absorption in rats fed a normal calcium diet. The stimulation of intestinal calcium absorption was associated with increased serum 1,25 dihydroxyvitamin D levels. An additive effect of vitamin K and vitamin D on intestinal calcium absorption was found only in rats fed a normal calcium diet. This study shows the differential effects of vitamin K and vitamin D supplementation on calcium balance in young rats fed a normal or low calcium diet. PMID- 16261993 TI - Dietary sesame seeds elevate alpha-tocopherol concentration in rat brain. AB - We have previously reported that dietary sesame lignan elevates alpha-tocopherol concentration and decreases lipid peroxidation in tissues and serum of rats fed alpha-tocopherol. In this study, the effect of dietary sesame seeds on alpha tocopherol concentration and lipid peroxidation in rat brain was examined. In experiment 1, male Wistar rats (4 wk old) were fed a vitamin E-free diet, or a diet containing alpha-tocopherol with or without sesame seeds for 1, 4 and 8 wk. The dietary sesame seeds elevated the alpha-tocopherol and lowered the thiobarbituric acid-reactive substance (TBARS) concentrations in the brain of the rats fed alpha-tocopherol for 4 and 8 wk. The dietary sesame seeds maintained the high alpha-tocopherol concentration in the brain during the experimental period, while the concentration of the rats fed alpha-tocopherol without sesame seeds was lowered after 8 wk. Then, the alpha-tocopherol concentration in various regions of the brain of rats fed a basal level of alpha-tocopherol with sesame seeds was compared with that of rats fed an excess amount of alpha-tocopherol in experiment 2. The alpha-tocopherol concentration in the cerebrum, cerebellum, brain stem and hippocampus of the rats fed 50 mg alpha-tocopherol/kg with sesame seeds was higher than those of the rats fed 500 mg alpha-tocopherol/kg without sesame seeds. These results suggest that the dietary sesame seeds are more useful than the intake of an excess amount of alpha-tocopherol, for maintaining a high alpha tocopherol concentration and inhibiting lipid peroxidation in the various regions of the rat brain. PMID- 16261992 TI - Influence of phospholipids on beta-carotene absorption and conversion into vitamin A in rats. AB - This study determines the effect of lysophosphatidylcholine (lysoPC) and phosphatidylcholine (PC) in mixed micelles on beta-carotene and retinyl palmitate levels in rats in order to delineate the role of micellar phospholipids in the intestinal uptake of beta-carotene and its conversion into vitamin A. The rats were fed a single dose of beta-carotene solubilized in lysoPC (LPC group), PC (PC group) or no phospholipids (NoPL, control group) in micellar form. The level of beta-carotene and retinyl palmitate in plasma and beta-carotene in liver was analyzed by HPLC after 1, 2, 3, 6 and 9 h of feeding. The postprandial levels of beta-carotene in plasma (599.9 pmol/mL, Area Under Curve (AUC)) and in liver (1,161.3 pmol/g) were significantly (p<0.05) higher in the LPC group compared with its level in plasma (207.2 pmol/mL) and in liver (616.5 pmol/g) of the PC group and in plasma (119.1 pmol/mL) and in liver (626.2 pmol/g) of the NoPL group. No difference was seen between the PC and NoPL groups. The results demonstrate that beta-carotene absorption and its accumulation in plasma and liver were unaffected by PC compared with NoPL, while lysoPC not only enhanced its accumulation but also increased cleavage of intestinally absorbed beta carotene into vitamin A as the AUC of plasma BC was higher and the AUC of retinyl palmitate in plasma of the lysoPC group was significantly higher than those of the other two groups. The results suggest that the luminal hydrolysis of PC to lysoPC is necessary for intestinal uptake of beta-carotene solubilized in mixed micelles. PMID- 16261994 TI - Effects of economic status and education level on the height and weight of community adolescents in Nepal. AB - There is scarce information on the relative importance of socio-economic factors in determining the adolescent anthropometric measurements. The objective of this study was to examine the effects of economic status, education level, and food consumption on the height and weight of community adolescents in Nepal. The study was done in the communities of the Kathmandu Valley area in Nepal. All together 426 unmarried adolescent girls aged 14-19 y were selected. The adolescents were interviewed regarding socioeconomic background (education, occupation and property possessions) and frequency of foods consumption. Height and weight were determined and BMI was calculated. Z-scores of height-for-age and weight-for-age were calculated based on the WHO/NCHS standard to avoid bias by age. The adolescents participating in the survey were categorized into three groups using the various indicators of economic status: Low Economic Status (LES) group, Middle Economic Status (MES) group and High Economic Status (HES) group. The Z scores of height and weight were significantly lower in the LES group than in the MES and HES groups (p<0.05). The Z-score of height was significantly increased with education level even under the condition of controlling economic level (p<0.05). Since the frequency of milk consumption was significantly related not only with height (p<0.05), but also with economic (chi2=31.6, df=4, p<0.001) and education levels (chi2=22.4, df=6, p<0.01), the increased height in the groups of the better economic status or the better education level was interpreted to be due to the outcome of the higher frequency of milk consumption. This study indicated that education was a more important factor affecting the height of the adolescents via improved food habits even under adverse economic conditions. PMID- 16261995 TI - Fat-soluble and water-soluble vitamin contents of breast milk from Japanese women. AB - To determine the concentrations of fat-soluble and water-soluble vitamins in the maternal milk of Japanese women, we collected human milk samples from more than 4,000 mothers living throughout Japan between December 1998 and September 1999, and defined as group A the 691 samples among these that met the following conditions: breast milk of mothers who were under 40 y of age, who did not smoke habitually and/or use vitamin supplements, and whose babies showed no symptoms of atopy and had birth weights of 2.5 kg or more. We then analyzed the contents of vitamins individually. Large differences were observed among the contents of individual human milk samples. The mean contents of each component were as follows: vitamin A, 159.0 +/- 95.2 IU/100 mL; vitamin E, 0.325 +/- 0.165 alpha-TE mg/100mL; vitamin D3 (cholecalciferol), 8.0 +/- 10.7 ng/100mL; vitamin B1 (thiamin), 12.3 +/- 3.2 microg/100 mL; vitamin B2, 38.4 +/- 12.7 microg/100 mL; vitamin B6, 5.7 +/- 2.5 microg/100 mL; vitamin B12, 0.04 +/- 0.02 microg/100 mL; vitamin C, 5.1 +/- 1.9 mg/100 mL; biotin, 0.50 +/- 0.23 microg/100 mL; choline, 9.2 +/- 1.8 mg/100 mL; folic acid, 6.2 +/- 2.9 microg/100 mL; inositol, 12.6 +/- 3.6 mg/100 mL; niacin (nicotinamide), 32.9 +/- 20.4 microg/100 mL and pantothenic acid, 0.27 +/- 0.09 mg/100 mL. The concentrations of derivatives and/or related compounds of vitamin A (retinol, beta-carotene), vitamin E (alpha-, beta-, gamma , and delta-tocopherol), and B2 (riboflavin, FMN, and FAD) were determined separately. The contents of each were found to vary greatly as the duration of lactation increased. The present results indicate that it is necessary to evaluate individual differences in human milk in order to perform valid research regarding infant formula. PMID- 16261996 TI - Effects of fasting and refeeding on expression of proteolytic-related genes in skeletal muscle of chicks. AB - This experiment was conducted to study the effects of fasting and refeeding on proteolytic-related gene expression in skeletal muscles of chicks. Chicks were fasted for 24 h, and refed for 2 h. Plasma Ntau-methylhistidine concentration, as an index of myofibrillar protein degradation, was increased by fasting, and that increment was reduced by refeeding. We also examined the expression of the protease mRNAs (calpain, proteasome, cathepsin and caspase-3) by real-time PCR of cDNA in skeletal muscles of fasting and refeeding chicks. Calpain (m-, mu-, and p94/calpain-3) mRNA expressions were also increased by fasting, and their increment was reduced by refeeding. Ubiquitin and 20S proteasome alpha subunit (alpha6 and alpha7) mRNA expressions as well as cathepsin B, and caspase-3 mRNA expression were likewise increased by fasting, with their increment also reduced by refeeding. These results indicate that fasting stimulates proteolytic-related gene expression, resulting in an increase in myofibrillar protein degradation, and that refeeding suppresses proteolytic-related gene expression, resulting in a decrease in myofibrillar protein degradation in chicks. PMID- 16261997 TI - Soy isoflavone supplementation alleviates oxidative stress and improves systolic blood pressure in male spontaneously hypertensive rats. AB - The aim of this study was to investigate the protective effect of isoflavone against hypertension, via the mitigation of oxidative stress and prevention of nitric oxide (NO, a potent vasodilator) reduction, in spontaneously hypertensive rats (SHR). The 8 wk-old male SHR were divided into two groups, and fed a casein based high fat diet (120 g fat, 1 g cholesterol/kg diet) for 30 d, either with or without 10 g of soy powder (containing 31.2% of isoflavones)/kg. During the 30-d study period, tail systolic blood pressures (BP) in the control SHR group increased, from 162.4 +/- 2.3 to 177.9 +/- 5.4 mmHg (p<0.05), while the isoflavone-supplemented group benefited from a clear antihypertensive effect (160.1 +/- 1.8 to 160.2 +/- 4.9 mmHg). The serum NO and total radical trapping antioxidant potential (TRAP) were elevated in the isoflavone group. The isoflavone group also experienced a significant decrease in oxidative DNA damage in leukocytes, using comet assay. DNA damage correlated positively with incremental BP during the study, and systolic BP at the end of the study (p<0.01). Our results indicate that soy isoflavone has an antihypertensive effect, possibly through the amelioration of oxidative stress, and the augmentation of NO production, in SHR. PMID- 16261998 TI - Establishment and issues of new educational system by nutrition teachers. AB - The Japanese school lunch system, which has over 100 y of history, has gained world attention for its well-developed and steady system, healthy Japanese-style menus including rice, and educational programs utilizing the school lunch system. Meanwhile, risk factors of health among students have been continuously worsening for the past dozen years or so. Therefore, Japan had hastened to develop better educational programs utilizing the school lunch and nutritional teachers. Under such circumstances, the Japanese Diet passed a bill to amend the School Education Law (hereinafter called the Amendment) which includes the establishment of an educational system by nutrition teachers in May, 2004. This system will be enforced in April, 2005. This system by nutrition teachers in Japan is well organized in both the legal and administrative senses and is attracting domestic and international attention as an example of an educational measure to improve the dietary life of students. The reason for this article in English is the additional intention of enlightening other countries by showing the political promotion problems of educational administration agencies, educational contents of nutrition teachers and some issues of the new educational system by nutrition teachers. PMID- 16261999 TI - Positive correlation between dietary intake of sodium and balances of calcium and magnesium in young Japanese adults--low sodium intake is a risk factor for loss of calcium and magnesium--. AB - The content of calcium (Ca) and magnesium (Mg) in sweat during exercise is considerably higher during a relatively low intake of sodium (Na) of 100 mmol/d than with an intake of 170 mmol/d. For this reason and also because Ca and Mg have a negative balance with a Na intake of 100 mmol/d, we analyzed the relationship between Na intake and balances of Ca and Mg in data from 11 balance studies. From 1986 to 2000, 109 volunteers (23 males, 86 females) with an age range of 18 to 28 y took part in mineral balance studies. The balance periods ranged from 5 to 12 d. In a given experiment, the diet of each subject contained the same quantity of food, although this varied between experiments, and was supplied during the balance period without consideration of body weight. In the data of all the studies (n= 109), the balances of Ca and Mg did not correlate positively with Na intake. However, when the data of the highest Na study were excluded, the balances of Ca and Mg correlated positively with Na intake. The mean value for the regression equation between Na intake and Ca and Mg balances when the respective balance was equal to zero were, 63.308 mg Na/kg BW/d (Ca: n=96, r2=0.134) and 60.977 mg Na/kg BW/d (Mg: n=96, r2=0.268), respectively. These values are considerably higher than Na requirements estimated by inevitable Na loss. Low dietary Na may therefore be a risk factor for maintaining positive balances of Ca and Mg. PMID- 16262000 TI - Regulatory mechanisms of 6,7-dimethyl-8-ribityllumazine formation in resting cells of a riboflavin-adenine-deficient mutant of Bacillus subtilis. AB - The regulatory mechanisms of riboflavin biosynthesis in the resting cells of a riboflavin-adenine-deficient mutant of Bacillus subtilis were examined. The growth and pH of the medium remained unchanged in spite of the administration of 0-200 microg/mL riboflavin to the medium during incubation of the resting cells for 17 h. However, the formation of 6,7-dimethyl-8-ribityllumazine (DMRL) and flavin adenine dinucleotide (FAD) was restricted and augmented and then reached its plateau, showing an inverse relation in the addition of riboflavin up to 50 microg/mL to the medium and a parallel relation in the supplementation of riboflavin up to 200 microg/mL to the medium. In the experiments, the amount of flavin mononucleotide (FMN) was negligible and riboflavin was not detected in the resting cells. The results indicated that not the repression of related enzymes but negative feedback inhibition by FAD, but not that by riboflavin or FMN, is operative in the biosynthetic pathway of riboflavin in the riboflavin adenine double-less mutant of Bacillus subtilis. PMID- 16262001 TI - Molecular cloning and analysis of the 5'-flanking region of the human thiamin pyrophosphokinase gene. AB - We cloned and analyzed the 5'-flanking region of the human thiamin pyrophosphokinase gene (hTPK1). Truncation analysis using transiently transfected HepG2 cells revealed the minimal region required for basal activity of the hTPK1 promoter, which was encoded in a sequence between -105 and +441 relative to the transcription start site. In an electrophoretic mobility shift assay using the nuclear extracts from HepG2 cells and the synthetic oligonucleotide containing the Sp1 site, specific DNA-protein complexes were identified. These findings indicate the importance of the Sp1 cis-element in regulating the hTPK1 gene expression. PMID- 16262002 TI - Protective effects of gamma-aminobutyric acid in rats with streptozotocin-induced diabetes. AB - The effects of gamma-aminobutyric acid (GABA) in rats with experimental diabetes mellitus were examined. Diabetes mellitus was induced in adult male Wistar rats by streptozotocin (STZ) injection. Oral administration of GABA (100 or 200 mg/kg body weight/d) for 10 d to the diabetic rats resulted in a significant decrease in their serum glucose level. GABA also reduced the level of glycosylated protein in serum, indicating an improvement of hyperglycemic conditions. Rats with STZ induced diabetes showed arrested body weight gain and an increase in both liver and kidney weight, whereas oral administration of GABA attenuated the organ hypertrophy induced by hyperglycemia. In addition, the degree of serum thiobarbituric acid (TBA)-reactive substance level was significantly lower in the rats treated with 100 mg GABA, and the degree of TBA-reactive substance in the liver and kidney was reduced by GABA in a dose-dependent manner. These results suggest that GABA treatment protects against the development of diabetic complications resulting from impaired glucose metabolism and enhanced oxidative stress. PMID- 16262003 TI - Attenuation of renal ischemia-reperfusion injury by proanthocyanidin-rich extract from grape seeds. AB - The effects of proanthocyanidin-rich extract in rats subjected to renal ischemia reperfusion were examined. Proanthocyanidin-rich extract, which is prepared from grape seeds (Vitis vinifera L.), was given orally at doses of 5 and 10 mg/kg body weight/d for 20 consecutive days prior to ischemia-reperfusion. Administration of proanthocyanidin-rich extract attenuated renal dysfunction, as indicated by serum urea nitrogen and creatinine levels. Additionally, in the ischemic-reperfused kidneys, increased levels of thiobarbituric acid (TBA)-reactive substance and alterations of antioxidant enzyme activities such as superoxide dismutase, catalase and glutathione peroxidase (GSH-Px) were observed. Proanthocyanidin-rich extract-treated groups showed significantly reduced renal TBA-reactive substance levels and enhanced catalase and GSH-Px activities. These results suggest that proanthocyanidin-rich extract has protective effects against ischemia-reperfusion induced renal damage associated with oxidative stress. PMID- 16262004 TI - Histochemical structure of the mucus gel layer coating the fecal surface of rodents, rabbits and humans. AB - The aim of this study was to investigate whether a mucus layer covers the surface of various animal feces and, if so, to show the structure and mucin composition of this layer. The freshly excreted feces of mice, rats, guinea pigs, rabbits and humans were fixed with Carnoy solution. Cross-sections approximately 5 microm thick were stained with alcian blue (AB) or the periodic acid Schiff (PAS) reaction. We measured the thickness of the mucus layer on the fecal surface of these sections. The fecal surface was covered with a continuous mucus layer in all specimens. The mucus layers of mice, rats and humans consisted of the alternate stratification of AB-positive and PAS-positive mucin layers. In contrast, the mucus layer consisted of an inner PAS-positive neutral mucin layer and an outer AB-positive acidic mucin layer in guinea pigs and rabbits. The average thicknesses of the mucus layers upon the fecal surfaces for mice, rats, guinea pigs, rabbits and humans were 19 +/- 12, 22 +/- 14, 15 +/- 4.6, 19 +/- 14 and 17 +/- 11 microm, respectively. These results demonstrated that the fecal surfaces are covered with continuous mucus layers in rodents, rabbits and humans, with substructures varying among species. PMID- 16262006 TI - Breakfast with Henry. PMID- 16262005 TI - Effect of dietary factors on incidence of type 2 diabetes: a systematic review of cohort studies. AB - We systematically reviewed cohort studies on the effect of nutrient and food intake (except for alcohol) on the incidence of type 2 diabetes, which had been published in English as of May 2004. Using the MEDLINE (PubMed) database as well as reference lists of searched papers, 15 individual cohort studies (a total of 31 papers) were identified. The number of subjects (n= 895-85,060), follow-up length (5.9-23 y), the number of diabetes cases (n= 74-4,085), dietary assessment method used (simple food questionnaire, food frequency questionnaire, food frequency interview, diet history interview, and 24-h recall), and method of case ascertainment (questionnaire, oral glucose tolerance test, fasting glucose level, death certificate, and nationwide registry) varied among studies. For nutrients, intakes of vegetable fat, polyunsaturated fatty acid, dietary fiber (particularly cereal fiber), magnesium, and caffeine were significantly inversely correlated and intakes of trans fatty acid and heme-iron, glycemic index, and glycemic load were significantly positively correlated with the incidence of type 2 diabetes in several papers. For foods and food groups, several papers showed significantly decreased risk for type 2 diabetes with the higher consumption of grain (particularly whole grain) and coffee, and significantly increased risk with processed meat consumption. Because all the studies were carried out in Western countries, however, research in non-Western countries including Japan is needed. PMID- 16262007 TI - Osteotomy about the knee: applications, techniques, and results. AB - Varus or valgus malalignment of the knee may be either a cause or a consequence of unicompartmental knee arthritis in young, active adults. Proximal tibial osteotomy for the varus knee and distal femoral osteotomy for the valgus knee have been used for decades to manage this condition; however, their use has decreased significantly in recent years as the popularity of unicompartmental and total knee arthroplasty has grown. With the advent of biologic resurfacing techniques for focal full-thickness articular cartilage injury, combined or staged high tibial osteotomy is becoming increasingly popular. In addition, in the face of cruciate ligamentous instability with or without posterolateral corner instability coupled with varus malalignment, high tibial osteotomy with and without ligament reconstruction provides a solution to complex orthopedic problems. Recent long-term follow-up studies have concluded osteotomy allows for improved function and pain relief in properly selected young patients. PMID- 16262008 TI - Total knee replacement for patellofemoral disease. AB - This study evaluates the technical results and outcomes of total knee replacement (TKR) in a group of patients with isolated patellofemoral disease based on the amount of bone loss and patella subluxation seen at the time of surgery. Thirty three TKRs in 25 patients comprised the study group. All patients had radiographic evidence of isolated end-stage disease in the patellofemoral joint and had cemented tri-compartmental knee replacements. All patellae could be resurfaced. Average follow-up was 5.2 years (range: 3.8-8.4 years). Average Knee Society Scores was 62 preoperatively and 96 at final follow-up. There was no correlation in outcomes related to either the amount of patellar wear, size of the patella, or amount of subluxation of the patella. Total knee replacement is an effective means of managing end-stage arthritis of the patellofemoral joint. PMID- 16262009 TI - Arthroscopic meniscal repair: analysis of treatment failures. AB - The rationale for meniscal repair is based on the importance of the meniscus in overall knee function and stability as well as the inferior results seen with meniscectomy. The high success rate usually seen with arthroscopic meniscal repair has made it the treatment of choice for peripheral meniscal tears. This study reviewed the records of patients who have failed meniscal repair surgery to gain greater understanding of the factors that may predispose a patient to a failed outcome. From 1987 to 2002, three hundred meniscal repairs were performed (203 medial and 97 lateral). Thirty-seven patients had failed meniscal repairs. Records were available for 33 (89%) patients. The mean patient age was 25 years (range: 13-48 years) at the time of meniscal injury. The average initial tear size was 2.7 cm with a mean rim width of 2.3 mm. Eighty-eight percent occurred in ACL tears. The average time interval from initial repair to the recurrence of symptoms was 34 months. Patients who were older at the time of meniscal repair failed significantly later than those patients who were younger at the time of repair. With age stratification, those patients who were aged > or =29 years at time of meniscal repair failed at an average of 23 months. In contrast, patients who were aged > or =30 years at the time of repair failed at an average of 53 months. Larger initial tears failed significantly sooner than smaller tears. Initial tears with larger rim widths demonstrated a trend toward shorter time to failure. Patients who underwent combined ligament reconstruction with meniscal repair failed at an average of 37 months. Deficient ACLs that were treated with isolated meniscal repairs (ACL intact) failed at an average of 16 months. PMID- 16262010 TI - Chondral injury after meniscal repair with rapidLoc. PMID- 16262011 TI - Bone and soft-tissue tumors about the knee. Foreward. PMID- 16262012 TI - Evaluation of masses around the knee. PMID- 16262013 TI - Complications of modular oncology knee prostheses for sarcomas about the knee. PMID- 16262014 TI - Reconstruction options for pediatric bone tumors about the knee. AB - The surgical decisions involved with high-grade sarcoma surgery about the knee in pediatric patients are complicated. The orthopedic oncologist must have experience as well as a great deal of insight into the intricacies of each procedure and the wishes and demands of patients--not only while they are young but in into their adult life. It is difficult to predict the interests and vocation young patients will choose as their lives progress. However, if the patient qualifies as a candidate for limb preservation, the functional and cosmetic results obtained by limb preservation seem to be preferred. Amputation and rotationplasty should be reserved for patients who are not candidates for limb preservation or who are in an otherwise salvage situation. The prognosis for quality of life and limb after treatment for bone sarcomas has increased dramatically during the past decade. Survival approaching the 70% to 90% range can be expected, with >90% of those patients having limb preservation surgery. With further advancements in the biologic control of tissue healing, bioengineering, and prosthetic design and implantation, the future of these patients continues to be optimistic. PMID- 16262015 TI - Going for growth. PMID- 16262016 TI - Perceptions of growth monitoring and promotion among an international panel of district medical officers. AB - The growth chart has been proposed as an educational tool to make the child's growth visible to both health workers and caregivers and to enhance communication between them. In the case of growth faltering, this would trigger timely corrective measures. Although the relevance of growth monitoring and promotion (GMP) has often been questioned in the literature, opinions of District Medical Officers responsible for local implementation of GMP are unknown. The aim of this qualitative research was to explore the perceptions and difficulties of an international panel of District Medical Officers regarding GMP. As an exploratory study, in-depth interviews of an international panel of District Medical Officers (n=19) were conducted. Data were coded using the QSR Nudist 5.0 software. A discrepancy between intended purposes and practice of GMP was detected at two levels. First, lack of participation of care-givers was reported. Second, the District Medical Officers expressed a restrictive interpretation of the concept of growth monitoring. The communication with parents was never reported as a means or a result of GMP, neither as an evaluation criterion of programme efficiency. The growth chart was mainly considered a tool intended to be used by health services for the purpose of diagnosis. This two-fold discrepancy between the intention of international policy-planners and practice of local programme implementers could be a crucial factor affecting the performance of GMP. More emphasis should be put on social communication and involvement of caregivers. PMID- 16262017 TI - Acceptability of and adherence to dispersible zinc tablet in the treatment of acute childhood diarrhoea. AB - Zinc treatment is now recommended by the World Health Organization as part of the routine management of acute childhood diarrhoea. A dispersible zinc tablet formulation was developed taking into account the taste, cost, and feasibility to distribute and store. Only limited information is available on the acceptability of and adherence to dispersible zinc tablet. No study has formally assessed whether the formulation is acceptable to children and if caretakers can adhere to the instructions regarding preparation, dosage, and duration of treatment. This community-based study aimed at determining the acceptability of and adherence to a dispersible zinc tablet formulation in a cohort of children (n=320) aged less than five years. Caretakers of children with acute childhood diarrhoea were prescribed zinc tablet treatment and followed up after 2-3 weeks. The formulation was acceptable to children; 90.1% of 303 caretakers perceived that the tablets were equally or even more acceptable to their children compared to other medicines. Ninety-eight percent of the children received the standard dose of one tablet per day, and 55.8% completed the full 10-day course of zinc treatment. Adherence rates did not vary by age or gender of the child. These findings indicate that the tablet formulation is acceptable, but further efforts are required to enhance adherence. PMID- 16262018 TI - Feeding practices and factors contributing to wasting, stunting, and iron deficiency anaemia among 3-23-month old children in Kilosa district, rural Tanzania. AB - Infants in Tanzania are particularly vulnerable to under-nutrition during transition from breastmilk (as the only source of nourishment) to solid foods. A cross-sectional study was undertaken in Kilosa district in Tanzania to determine the feeding practices and the extent of wasting, stunting, and iron-deficiency anaemia. The study was done in two stages: in the first stage, a 24-hour dietary assessment was conducted to identify the type of complementary foods given and the eating habits according to age for 378 children aged 3-23 months. In the second stage, a progressive recruitment of 309 infants aged six months was made to measure weight, length, haemoglobin (Hb) concentration, zinc protoporphyrin concentration, and malaria parasitaemia. Birth-weight, the potential contributing factor to under-nutrition and iron-deficiency anaemia, was obtained from the children's clinic cards. The 24-hour dietary assessment revealed that children consumed mainly a thin porridge prepared from maize flour as complementary food. Carbohydrates contributed most energy (on average 69%), followed by fats (18.6%) and protein (on average 12.1%). The complementary food co-vered only 15%, 20%, and 27% of the recommended iron intake for children aged 6-8, 9-11 and 12-23 months respectively. The mean Hb concentration was 9.3 +/- 1.9 g/dL, 68% of the infants were moderately anaemic (7 < or =11 g/dL), and about 11% were severely anaemic with Hb below 7 g/dL, while 21% were non-anaemic Hb (> or =11 g/dL). Equally, the mean zinc protoporphyrin concentration was 10.0 +/- 6.2 microg/g Hb, and 76% of the infants were iron-deficient (>5 microg/g Hb). The prevalence of stunting was 35%, while wasting was only 1.3%. Low birth-weight and low body mass index of mothers were the strong predictors of stunting, whereas low birth-weight and iron-deficiency were the strong predictors of anaemia. The prevalence of malaria parasitaemia was high, affecting 50% of the infants. Having malaria was the only independent predictor associated with stunting, anaemia, and iron deficiency. There is an urgent need to improve tradi-tional complementary foods in the studied community in terms of energy density, amount of fat in the diet, and bioavailability of macro and micronutrients. PMID- 16262019 TI - Epidemiology of HIV infection among brothel-based sex workers in Kolkata, India. AB - A cross-sectional study was carried out during January-June 2004 to assess the status of HIV infection among brothel-based sex workers in Kolkata city, India. Six hundred and twenty-two sex workers, selected from six brothels, were included in the study to assess their HIV status, and 362 sex workers, a subset of the above population, were interviewed to study their risk behaviour and practices. Blood samples were collected from each sex worker for testing HIV by an unlinked anonymous method. The prevalence of HIV infection was 9.6%, but was much higher among younger sex workers aged 20 years or less (27.7%) compared to the older age group (8.4%). This difference was statistically significant, indicating an association of younger ages with HIV infection [p = 0.006 and odds ratio (OR) = 4.18; (95% confidence interval (CI) 1.2-13.8)]. HIV was not associated with duration of sex work, average number of clients entertained per day, condom use, practice of sex during menstruation, pre-coital examination of penis for visible ulcer/discharge, suffering from sexually transmitted infections, and entertaining clients outside the brothel. There was a big gap between the reported and the evaluated condom use by sex workers. The results suggest that there is a need to develop suitable HIV intervention strategies, considering the socioeconomic and cultural aspects of city sex workers, with a provision for continuous monitoring and evaluation. PMID- 16262020 TI - Routine use of antimicrobials by pregnant Indian women does not improve birth outcome: a randomized controlled trial. AB - Low birth-weight is a leading health problem in developing countries. In a randomized controlled trial, the effect of antimicrobials in pregnant women on improving birth-weight and duration of gestation was evaluated. Two hundred twenty-four pregnant women in their second trimester were randomized to receive metronidazole (200 mg 3 times daily for 7 days) and cephalexin (500 mg twice daily for 5 days) orally by one group. The mean (+/-SD) birth-weights were 2,545 g (+/-374) and 2584 g (+/-358, p=0.51), the low birth-weight rates (<2.5 kg) were 40% and 36% (p = 0.28), and the prematurity rates were 8% and 11% (p = 0.6) in the treated group and the control group respectively. Due to small sample size, it is cautiously concluded that routine antimicrobials for genital and urinary tract infections of pregnant women do not improve birth-weight or duration of gestation. Rather an unexpected observation was the proportion requiring caesarian section or forceps, which was five-fold higher in the treated group (p = 0.001), and given no plausible explanations, this finding needs confirmation. Stunted mothers (<25th centile or 146.4 cm) had two-fold higher risk for low birth-weight (p = 0.04) and assisted delivery (p = 0.1). Low maternal body mass index (<25th centile or 18) had six-fold higher risk for stillbirth or abortion (p = 0.007), and high body mass index (>75th centile or 21.2) had three-fold higher risk for assisted delivery (p = 0.003). PMID- 16262021 TI - Epidemiological aspects of rotavirus infection in Ahwaz, Iran. AB - Rotavirus is the major cause of diarrhoea in children worldwide. In this study, conducted in the city of Ahwaz, Iran, during November 2001-March 2002, stool samples from 200 inpatient (n=63) and outpatient (n=137) children aged 1-24 month(s) were analyzed. Polyacrylamide gel electrophoresis was used for isolating rotavirus. Rotavirus was isolated from 36 (26.3%) of the 137 stool samples of outpatients and from 23 (36.5%) of the 63 stool samples of inpatients. The overall frequency of rotavirus in this population was 29.5%. The highest detection of rotavirus was made in children aged 7-12 months, which demonstrated that the relationship between age and rate of rotaviral infection was statistically significant (p < 0.05). The predominant electrophoretic pattern detected was the long (L) electrophoretype (46 of 59; 78%), followed by the short (S) electrophoretype (12 of 59; 20.3%). One strain had a mixed pattern. Such analysis throughout Iran would assist in developing sound guidelines for the prevention of rotavirus infections. PMID- 16262022 TI - Growth patterns of Qatari school children and adolescents aged 6-18 years. AB - The study was conducted to analyze the patterns of growth in height and weight and the prevalence of over-weight among Qatari school children aged 6-18 years. Weights and heights of a cross-sectional sample of Qatari school children were measured. These children were selected randomly, in equal proportions of age and gender, from different schools from urban and semi-urban districts. Appropriate statistical procedures were performed to produce smooth percentile curves for boys and girls using a two-stage approach. Initial curve smoothing for selected major percentiles was accomplished by various paramet-ric and non-parametric procedures. In the second stage, a normalization procedure was used for creating z-scores that closely matched the smooth percentile curves. The height and weight results were compared with the international reference values of National Center for Health Statistics/Centers for Disease Control and Prevention (NCHS/CDC). The prevalence of over-weight was calculated using the new International Obesity Task Force (IOTF) reference. Of 7442 Qatari children studied, 50.3% were male and 49.7%' female. The mean values for height, weight, and body mass index (BMI) increased with the age for both boys and girls until the age of 18 years, except BMI, which stabilized at the age of 16-18 years at around 22.6 for boys and at 21.6 for girls. The growth patterns of the Qatari children, aged 6-18 years, appeared to be comparable with those of the NCHS/CDC reference. The weight-for age centile curves of the Qatari boys tended to be superior to those of the NCHS/CDC reference until the age of 15 years, less so those of the Qatari girls. In contrast, the height-for-age centile curves of the Qatari children tended to deviate in a negative sense from the NCHS/CDC reference curves, for boys and girls from age around 11 years and 13 years respectively. The deviation of the smoothed median height-for-age curves from the reference in adolescence could most likely be attributed to a later maturation among the Qatari children. The prevalence of under-weight, over-weight, and obesity for the Qatari children was quite below the CDC and IOTF rates, except for girls aged 6-9 years. More males than females were over-weight or obese according to either the local, the CDC, or the IOTF reference, and the prevalence increased with age. A good percentage of the Qatari children was at risk of being over-weight, which needs more attention because the development of obesity results in different types of diseases associated with changes in body composition. PMID- 16262023 TI - Risk factors for mortality due to shigellosis: a case-control study among severely-malnourished children in Bangladesh. AB - To determine the risk factors for death of severely-malnourished Bangladeshi children with shigellosis, a case-control study was conducted at the Clinical Research and Service Centre of ICDDR,B: Centre for Health and Population Research in Dhaka, Bangladesh. One hundred severely-malnourished children (weight-for-age <60% of median of the National Center for Health Statistics), with a positive stool culture for Shigella dysenteriae type 1 or S. flexneri, who died during hospitalization, were compared with another 100 similar children (weight-for-age <60% and with S. dysenteriae type 1 or S. flexneri-associated infection) discharged alive. Children aged less than four years were admitted during December 1993-January 1999. The median age of the cases who died or recovered was 9 months and 12 months respectively. Bronchopneumonia, abdominal distension, absent or sluggish bowel sound, clinical anaemia, altered consciousness, hypothermia, clinical sepsis, low or imperceptible pulse, dehydration, hypoglycaemia, high creatinine, and hyperkalaemia were all significantly more frequent in cases than in controls. In multivariate regression analysis, altered consciousness (odds ratio [OR]=2.6, 95% confidence interval [CI] 1.0-6.8), hypoglycaemia (blood glucose <3 mmol/L (OR=7.8, 95% CI 2.9-19.6), hypothermia (temperature <36 degrees C) (OR=5.7, 95% CI 1.5-22.1), and bronchopneumonia (OR=2.5, 95% CI 1.1-5.5) were identified as significant risk factors for mortality. Severely-malnourished children with shigellosis having hypoglycaemia, hypothermia, altered consciousness and/or bronchopneumonia were at high risk of death. Based on the findings, the study recommends that early diagnosis of shigellosis in severely-malnourished children and assertive therapy for proper management to prevent development of hypothermia, hypoglycaemia, bronchopneumonia, or altered consciousness and its immediate treatment are likely to reduce Shigella-related mortality in severely-malnourished children. PMID- 16262024 TI - Availability and use of emergency obstetric care services in four districts of West Bengal, India. AB - Process indicators have been recommended for monitoring the availability and use of emergency obstetric care (EmOC) services. A health facility-based study was carried out in 2002 in four districts of West Bengal, India, to analyze these process indicators. Relevant records and registers for 2001 of all studied facilities in the districts were reviewed to collect data using a pre-designed schedule. The numbers of basic and comprehensive EmOC facilities were inadequate in all the four districts compared to the minimum acceptable level. Overall, 26.2% of estimated annual births took place in the EmOC facilities (ranged from 16.2% to 45.8% in 4 districts) against the required minimum of 15%. The rate of caesarean section calculated for all expected births in the population varied from 3.5% to 4.4% in the four districts with an overall rate of 4%, which is less than the minimum target of 5%. Only 29.9% of the estimated number of complications (which is 15% of all births) was managed in the EmOC facilities. The combined case-fatality rate in the basic/comprehensive EmOC facilities was 1.7%. Major obstetric complications contributed to 85.7% of maternal deaths, and pre-eclampsia/eclampsia was the most common cause. It can be concluded that all the process indicators, except proportion of deliveries in the EmOC facilities, were below the acceptable level. Certain priority measures, such as making facilities fully functional, effective referral and monitoring system, skill based training, etc., are to be emphasized to improve the situation. PMID- 16262025 TI - A comparison of two systems for chlorinating water in rural Honduras. AB - This study investigated a small subset of the two community water-disinfection systems--hypochlorinators and tablet feeders-in rural Honduras. Levels of residual chlorine were assessed at three locations within the distribution system: the tank, the proximal house, and the distal house. The levels of residual chlorine were compared with the standard guidelines set by the Pan American Health Organization and the International Rural Water Association for potable water that require a minimum of 1.0 (tank), 0.5 (proximal house), and 0.2 (distal house) ppm for each location. The levels of residual chlorine were also compared across systems, e.g. hypochlorinators to tablet feeders. At the tank and proximal house, tablet feeders had significantly higher mean values for levels of residual chlorine (measured in ppm) than hypochlorinators (tank: 1.20 vs 0.67; proximal house: 0.44 vs 0.32, p < 0.001 for both) with no significant difference at the distal house (0.16 vs 0.16). At the tank and proximal house, tablet feeders were more likely to meet recommended standards than hypochlorinators (90.3% vs 13.3%, p < 0.0001 and 41.3% vs 23.7%, p < 0.0001) with a smaller difference seen at the distal house (30.6% vs 27.1%, p = 0.24). The apparent dichotomy in chlorine levels of tablet feeders (e.g. between tank/proximal house and distal house) is discussed. The results suggest that tablet feeders may be more effective than hypochlorinators in supplying clean water in rural, resource poor settings and possibly serve as an alternative technology for water disinfection. Further research on techniques for empowering and building capacity within community water boards will help organize and introduce sustainable water systems in developing countries. PMID- 16262026 TI - Impact of training on assessment of diarrhoea and acute respiratory infection at government health facilities in Egypt. AB - Egypt began training of physicians in case management of diarrhoea and acute respiratory infection (ARI) in the 1980s and 1990s respectively. Data from an independent assessment demonstrate the impact of training in the use of clinical practice guidelines on the quality of clinical examinations of 579 children presenting with diarrhoea or ARI. These examinations were conducted by 115 government physicians in 80 government health facilities and in two Egyptian governorates. The quality of care, although better than that in the past, remains poor, and training had little effect. Poor examinations were related to misclassification and inappropriate treatment of serious illnesses. Such errors have potentially serious consequences. Egypt is now implementing the Integrated Management of Childhood Illness strategy. This effort should be independently assessed to measure the improvement in the quality of care provided by government health facilities in Egypt. PMID- 16262028 TI - Microbial contamination of herbal preparations in Lagos, Nigeria. PMID- 16262027 TI - Entamoeba moshkovskii and Entamoeba dispar-associated infections in pondicherry, India. AB - The prevalence of Laredo strain--Entamoeba moshkovskii--and non-pathogenic E. dispar in patients attending the Jawaharlal Institute of Postgraduate Medical Education and Research hospital, Pondicherry, India, is reported here. E. moshkovskii is reported for the first time in India. The species are morphologically indistinguishable from pathogenic E. histolytica. Of 746 stool samples screened, 68 showing cyst or trophozoite stage of E. histolytica, E. dispar, or E. moshkovskii were subjected to small subunit (SSU) rRNA gene-based polymerase chain reaction, which revealed a higher prevalence of E. dispar (8.8%) and E. moshkovskii (2.2%) compared to E. histolytica (1.7%) in patients. Only 19% of the 68 stool samples, resembling E. histolytica by microscopy, were actually E. histolytica, implying that 81% of suspected infections were misdiagnosed and would have been treated unnecessarily with anti-amoebic drugs. PMID- 16262029 TI - Awareness of HIV/AIDS and risky sexual behaviour among male drug users of higher socioeconomic status in Dhaka, Bangladesh. PMID- 16262030 TI - Write it down! PMID- 16262031 TI - Periodontal disease and general health. AB - There is considerable interest in the possible relationship between periodontal disease and pregnancy complications, cardiovascular disease, diabetes and respiratory diseases. This interest has been stimulated by data obtained from epidemiological studies and a better understanding of the pathogenesis of periodontal disease. Whilst much of the data support an association between oral and general health, further studies are needed to accept or reject a causal relationship. PMID- 16262032 TI - Trends in indirect dentistry: 6. Provisional restorations, more than just a temporary. AB - The provision of well-fitting, functional provisional restorations is important for a wide variety of reasons, including maintenance of the stability of inter- and intra-arch relationships and positional stability of prepared teeth, and the preservation of occlusal function of anterior provisional restorations by providing appropriate protrusive and lateral guidance. Provisional restorations should be of sufficient strength to resist the forces of occlusion and should be luted with a cement that will resist the forces of removal, yet allow easy removal at the fit appointment without leaving a residue on the prepared tooth. This paper describes the use of provisional restorations in indirect restorative dentistry and suggests some specific techniques for clinical use. PMID- 16262033 TI - The role of the dental team in preventing and diagnosing cancer: 5. Alcohol and the role of the dentist in alcohol cessation. AB - Alcohol use contributes to many health disorders and social problems which may affect both the individual and the community, and is a major risk factor for oral cancer and potentially malignant lesions - leukoplakia and erythroplakia. Counselling by doctors and dentists can increase users' motivation to stop alcohol use but is not often applied in a systematic or frequent manner to people presenting with potentially malignant oral lesions. This paper makes recommendations for interventions by health professionals to encourage and aid cessation of alcohol use as a part of prevention of oral cancer. PMID- 16262034 TI - An overview of tooth discoloration: extrinsic, intrinsic and internalized stains. AB - The causes of tooth discoloration are varied and complex but are usually classified as being either intrinsic, extrinsic or internalized in nature. Dietary chromogens and other external elements deposit on the tooth surface or within the pellicle layer either directly or indirectly to form extrinsic discoloration. Stains within the dentine or intrinsic discoloration often results from systemic or pulpal origin, while internalized stains are the result of extrinsic stains entering the dentine via tooth defects such as cracks on the tooth surface. PMID- 16262035 TI - The medical management and dental implications of long QT syndrome. AB - Long QT is a disorder of the heart's conduction system. It can be congenital or acquired. The acquired type is caused mainly by medications. The diagnosis of LQT depends on the clinical features, the family history, and the ECG findings. The clinical presentations range from dizziness to syncope and sudden death. The dentist should consult the cardiologist to avoid any complications that may put the LQTS patient at risk. Medications such as erythromycin and antifungals are contra-indicated and should not be prescribed. Local anaesthetics containing adrenaline and bupivacaine should be avoided. Stress can precipitate symptoms and lead to sudden death. PMID- 16262036 TI - Dental management of patients taking methadone. AB - Methadone is a synthetic opiate used in the treatment of opiate addiction. Various side-effects have been associated with the use of methadone. These include xerostomia, which can contribute to a high caries rate. The UK Regional Drug Misuse Database reported that around 118,500 drug users were receiving treatment from drug misuse agencies and GPs. The vast majority (87%) were receiving treatment from community specialist services. As many drug abusers have poor oral health, general dental practitioners are likely to encounter such individuals. It is essential therefore that dentists are aware of the potential difficulties that may be encountered when treating subjects receiving methadone. These problems may relate to previous drug abuse and to the effects of methadone therapy. PMID- 16262037 TI - Dentistry and homeopathy: an overview. AB - For the dentist who endeavours to practise as a physician of the mouth with preventive dental medicine as a part of his/her service, clinical experience has demonstrated that homeopathy can be of benefit. Many homeopathic remedies have been found to be effective for dental conditions, namely dental caries, dental abscess, oral lesions, post-extraction bleeding and even medications to treat the anxious and nervous child. PMID- 16262038 TI - Physical signs for the general dental practitioner. PMID- 16262039 TI - Delegate clerical functions and free up professionals' time. PMID- 16262040 TI - CM, social work duties handled by specialists. PMID- 16262041 TI - CMs face challenges of hurricane Katrina. PMID- 16262043 TI - Salaries are up, but CMs are putting in long hours. PMID- 16262042 TI - Initiative helps hospital improve quality indicators. PMID- 16262044 TI - System aims to improve patient throughput. PMID- 16262045 TI - [Net primary productivity of several mangrove species under controlled habitats]. AB - Three mangrove species Sonneratia caseolaris, Kandelia candel and Aegiceras corniculatum were planted in different fishponds in April 2002 with three planting--breeding area proportions of 45 : 55, 30 : 70 and 15 : 85, respectively, and the growth of test mangrove plants were surveyed during 2 years after planting. The results showed that S. caseolaris and A. comiculatum could grow well in the coupling system, while K. candel could not. The survival percentage of S. caseolaris, K. candel and A. corniculatum was 92.9%, 93.9% and 44.7%, respectively. During the 2 years, the height of S. caseolaris increased 457.0 cm, and its basal diameter increased from 12.6 mm to 98.7 mm. A. corniculatum had an increment of 26.1 cm in height and 36.5 mm in basal diameter, while K. candel only had an increment of 20.4 cm in height and 26.4 mm in basal diameter. Based on the height, basal diameter, and biomass of trunk, branch, leaf and root, regressive equations of the four organs' biomass were obtained, and the whole biomass of standing trees in the coupling system was calculated. The average biomass of S. caseolaris was 5 597.8 g x m(-2) in April 2004, being increased 5 559.5 g x m(-2) in 2 years. At the same period, the standing biomass of A. corniculatum and K. candel was 962.5 g x m(-2) and 66.0 g x m(-2), with an increase of 932.7 g x m(-2) and 57.0 g x m(-2), respectively. The biomass of plant organs was in the order of stem > branch > root > leaf for S. caseolaris, leaf > branch > stem > root for A. corniculatum, and stem > root > leaf > branch for K. candel. The litter fall production of mangrove plants in 2 years was 1 149.2 g x m(-2), 170.8 g x m(-2) and 7.1 g x m(-2) for S. caseolaris, A. corniculatum and K. candel, respectively. Leaf litter took up more than half of the whole litter fall. From April 2002 to April 2004, the net primary production of S. caseolaris, A. corniculatum and K. candel was 7 048.9 g x m(-2), 1 105.9 g x m(-2) and 93.0 g x m(-2), respectively. The litter fall production occupied 20.5% of the net primary production for S. caseolaris, 15.4% for A. corniculatum, and 7.6% for K. candel, which meant that high productivity was accompanied by high return rate. PMID- 16262046 TI - [Seasonal variation of Tamarix ramosissima and Populus euphratica water potentials in southern fringe of Taklamakan Desert]. AB - The measurement of the seasonal and diurnal variations of Tamarix ramosissima and Populus euphratica water potentials in the southern fringe of Taklamakan Desert indicated that there was no apparent water stress for the two species during their growth period, with little change of predawn water potential and some extent decrease of midday water potential. Irrigation once or thinning had no significant effects on the water status of the plants, while groundwater appeared to be a prerequisite for the survival and growth of these species. It is very important to ensure a stable groundwater table for the restoration of Tamarix ramosissima and Populus euphratica in this area. PMID- 16262047 TI - [Population characteristics of Coptis chinensis var. brevisepala in Anhui Province and its endangering mechanism]. AB - The study on the distribution pattern, population size, and age structure of C. chinensis var. brevisepala in Anhui Province showed that C. chinensis var. brevisepala mainly distributed in the southern mountain areas such as Mts. Jiuhuashan, Guniujiang, Huangshan and Qingliang Peak. The habitats had two kinds, i.e., under woods and beside water ditches, both of which were shady and humid, with loose and acidic soils containing abundant organic matter. The shading degree surpassed 70%, air humidity reached 70% approximately 90%, and soil water content was more than 30%. The distribution pattern of existing C. chinensis var. brevisepala populations was clustering, with illogical age structure and small population size. It was suggested that the populations of C. chinensis var. brevisepala in Anhui Province were in an unsteady stage. Serious destroy of natural habitat and excessive digging and gathering adult individuals were the main causes of the rapid decrease of the individuals in some populations, which threatened the existence of C. chinensis var. brevisepala populations seriously. PMID- 16262048 TI - [Effects of nitrate application on alleviating photosynthesis restriction of Cinnamomum burmannii leaves under elevated CO2 concentration and enhanced temperature]. AB - In this study, potted C. burmannii saplings were cultured in a top-closed chamber with elevated CO2 (+ CO2, 731 micromol x mol(-1)) and ambient CO2(CO2, 365 micromol x mol(-1)), and at diurnal temperature (day/night) 25/23 degrees C and 32/25 degrees C, respectively. The gas exchange, calculated photosynthesis parameter, Rubisco content, and activated state of Rubisco were examined. The results showed that under + CO2 and at 25/23 degrees C, the mean photosynthetic rate (Pnsat) of sapling leaves was 5.1% higher than that under + CO2 and at 32/25 degrees C. Temperature enhancement declined Pnsat, while nitrate addition increased it. Under + CO2, saplings had lower V(cmax) and J(max) at 32/25 degrees C than at 25/23 degrees C. Temperature enhancement under + CO2 also declined V(cmax) and J(max). Under + CO2, lower photorespiration rate (Rp) occurred in leaves, but temperature enhancement could increase Rp. Under + CO2, Rubisco content (NR) and its active site per unit leaf area (M) decreased as diurnal temperature changed from 25/23 degrees C to 32/25 degrees C. Meanwhile, nitrate addition could increase NR and M. It may be suggested that nitrate addition could alleviate the restriction of photosynthesis under elevated CO2 concentration and enhanced temperature. PMID- 16262049 TI - [Quantitative characteristics of soil aggregates under different vegetations in upper reach of Minjiang River]. AB - Quantitative analysis on the soil aggregates under dark coniferous forest, coniferous and broad-leaved mixed forest, fargesia under the gap of dark coniferous forest, and sclerophyllous oaks (Quercus semicarpifolia) at Wolong Natural Reserve in the upper reach of Minjiang River showed that wet-sieving soil aggregates were of logarithmic- normal distribution, and the geometric mean diameters were negatively correlated to geometric standard deviation. The aggregates under coniferous and broad-leaved mixed forest and sclerophyllous oaks had larger sizes than those under other vegetations. The range of fractal dimension of soil aggregates was 2.40 - 2.78, along with more aggregates less than 0.25 mm in size. The fractal dimension of soil aggregates under dark coniferous forest and fargesia were larger than that under other vegetations. The soil aggregates with 3 - 1 mm and 1 - 0.5 mm in size had a higher stability, while those with > 10 mm and 0.5 - 0.25 mm in size were in adverse. The aggregate stability index of soil under coniferous and broad-leaved mixed forest was the highest, followed by that under sclerophyllous oaks, fargesia under the gap of dark coniferous forest, and dark coniferous forest, which meant that coniferous and broadleaf mixed forest and sclerophyllous oaks were favorable for soil aggregate stability. Significant correlations were found among the three quantitative characteristics, which could be used to indicate the stability of soil aggregates. PMID- 16262050 TI - [Effects of replacing natural secondary broad-leaved forest with Cunninghamia lanceolata plantation on soil biological activities]. AB - This paper studied the effects of replacing natural secondary broad-leaved forest with Cunninghamia lanceolata plantation in the south, central and upstate areas of subtropical China on the changes of soil chemical and biological properties. The results showed that after replacing with Cunninghamia lanceolata plantation, the total organic carbon (TOC) content in surface soil decreased by 31.51% - 58.24%, and the contents of soil total N and P, pH value, C/N and C/P also decreased to different degree. Soil microbial amount was less than that under natural secondary broad-leaved forest, soil urease, invertase, catalase and dehydrogenase activities decreased, while soil polyphenol oxidase activity increased by 8% - 40%. The respiration rate of Cunninghamia lanceolata soil was 51.15% - 54.48% lower than that of natural secondary broad-leaved forest soil. The correlation between soil TOC and polyphenol oxidase activity was negative (R = - 0.723, n = 18), while those between soil TOC, N, P and other enzyme activities were positive. It could be concluded that replacing natural secondary broad-leaved forest with Cunninghamia lanceolata plantation worsened soil quality, and the loss of soil organic matter in Cunninghamia lanceolata plantation ecosystem might be one of the important factors resulted in the decrease of soil nutrients and enzyme activities. PMID- 16262051 TI - [Effects of different ground clearance on soil fertility of Chinese fir stands]. AB - The study on the soil physical properties, nutrient contents, microbial amounts and enzyme activities of clear cutting and controlled burning Chinese fir stands showed that in comparing with the control, soil non-capillary porosity of clear cutting stand increased by 23%, whereas soil natural water capacity and capillary moisture capacity decreased by 25%. In controlled burning stand, soil bulk density increased by 10%, while soil non-capillary porosity, natural water capacity and capillary moisture capacity decreased by 61%, 48% and 26%, respectively. The contents of soil organic matter, total N, total P and total K in clear cutting stand decreased by 14%, 14%, 35% and 22%, and in controlled burning stand, they decreased by 37%, 37%, 47% and 7%, respectively. Soil alkalized N and available K in clear cutting stand increased by 24% and 31%, respectively, but soil available P decreased by 15%. The contents of soil alkalized N, available P and available K in controlled burning stand decreased by 25%, 43% and 40%, respectively. In clear cutting stand, the amounts of soil bacteria, fungi and actinomyces increased by 1.4, 11.3 and 0.8 times, respectively, but in controlled burning stand, the amounts of soil bacteria decreased by 24%, while those of soil fungi and actinomyces increased by 5.0 and 0.5 times, respectively. The activities of soil urease, catalase and cellulase in clear cutting stand increased by 1.9, 1.6 and 2.1 times, and in controlled burning land, they were 35%, 90% and 106% of the control, respectively. Damp soil had higher contents of organic matter, total N and total P, whereas porous soil was favorable for the accumulation of alkalized N, available P and available K and for the increase of soil urease activity. The amount of soil fungi decreased with increasing soil capillary porosity. Aerated soil was favorable for the increase of soil catalase activity. PMID- 16262052 TI - [Ecological distribution patterns of soil microbes under artificial Eucalyptus grandis stand]. AB - In order to find out the ecological distribution patterns of soil microbes under artificial Eucalyptus grandis stand, the amount and distribution of soil bacteria, fungi and actinomycetes in Hongya County of Sichuan Province were investigated in 2004. The results showed that soil microbial population under Eucalyptus grandis stand varied with seasons, being the maximum in autumn, fewer in spring, and the minimum in summer. The numbers were decreased in order of bacteria > actinomycetes > fungi, which were mainly concentrated in 0-20 cm soil layer, and rapidly decreased with increasing soil depth. In 0-60 cm soil layer, the numbers of aerobic bacteria were 0. 31 x 10(6) - 14.39 x 10(6), actinomycetes were 0.06 x 10(6) - 0.79 x 10(6), fungi were 0.06 x 10(6) - 0.79 x 10(6), and anaerobes were 0.05 x 10(6) - 3.22 x 10(6) CFU x g(-1). Comparing with artificial C. glauca stand and farming land, Eucalyptus grandis stand had a larger number of soil microbes, suggesting that Eucalyptus grandis was benefit for soil microbial activity. The Simpson and Shannon-Wiener index of the physiological groups of bacteria was 0.773 and 1.896, respectively. PMID- 16262053 TI - [Spatial pattern relationships between emerald ash borer larvae and their natural enemies]. AB - The investigation on the spatial patterns of emerald ash borer (EAB) larvae, parasitoid Spathius agrili, and EAB-preying woodpecker pecks in ash forests showed that in horizontal direction, EAB larvae, parasitoid S. agrili and woodpecker pecks were in negative binomial distribution, while in vertical direction, EAB larvae were distributed on ash trunks from above ground to 3.37 m in height, with a peak around the 1.50 m. Their natural enemies also favored this range. PMID- 16262054 TI - [Growth characteristics of rock plant Pogonatherum paniceum]. AB - The study on the growth characters, above and underground biomass, and root-shoot ratio of Pogonatherum paniceum grown on three types of substrates and five types of habitats in Chongqing showed that there were significant differences in plant growth characters, including stalk basal diameter, plant canopy and height, root system area and depth, and maximum root length among five habitats. For shoot growth, though the plants grown on purple soil had the maximum stalk basal diameter (15.18 cm), canopy (3 086.77 cm2) and height (6.58 cm) while those grown on purple sandy rock had the minimum values (stalk basal diameter 10.89 cm, canopy 1 868.79 cm2, and height 60.75 cm). There were no significant differences among three types of substrates. As for root system, there were significant differences between sandy rocks and purple soil, the plants grown on sandy rocks having higher means of root system area (1 389.14 cm2 and 1 487.14 cm2) and maximum root length (45.83 cm and 39.24 cm) than those grown on purple soil (root system area 717.09 cm2, maximum root length 21.42 cm). The plants grown on sandy rock allocated more biomass to root system, and had higher root-shoot ratio, which was helpful for its fixation on rock and its endurance on the desiccation and arid of rock substrate. PMID- 16262055 TI - [Spatial variability of nutrients in cultivated soils of Xinhui District, Jiangmen City]. AB - Employing geostatistical methods and GIS technology, this paper studied the spatial distribution characteristics of pH, organic matter, CEC, total nitrogen, available phosphorus and slowly available potassium in cultivated soils of Xinhui District, Jiangmen City. All the test variables were normally or log normally distributed. Semivariogram analysis showed that soil nutrients were moderately spatially-dependent in a given spatial range, except that soil total nitrogen was strongly spatially-dependent. It was shown from Kriging analysis that soil pH and CEC was the highest in the northeast, and soil organic matter content was higher in the middle and northeast investigation region. The area with 1.5-2.0 g x kg( 1) soil total nitrogen content accounted for 75.7% of the investigated region, mainly distributed in the west and east, that with > 40 mg x kg(-1) soil available phosphorus content was accounted for 48.7%, mainly distributed in the west and northeast, and the area that slowly available potassium content was 160 350 mg x kg(-1) accounted for 48.1%, mainly distributed in the east, northeast and the middle. PMID- 16262056 TI - [Soft-ridged bench terrace design in hilly loess region]. AB - Reconfiguration of hillside field into terrace is regarded as one of the key techniques for water and soil conservation in mountainous regions. On slopes exceeding 30 degrees, the traditional techniques of terracing are difficult to apply as risers (i.e., backslopes), and if not reinforced, are so abrupt and easy to collapse under gravity alone, thus damaging the terrace. To improve the reconfiguration of hillside field into terrace, holistic techniques of soft ridged bench terrace engineering, including revegetation, with trees and planting grasses on riser slopes, were tested between 1997 and 2001 in Xiabiangou watershed of Yan' an, Shaanxi Province. A "working with Nature" engineering approach, riser slopes of 45 degrees, similar to the pre-existing slope of 35 degrees, was employed to radically reduce gravity-erosion. Based on the concepts of biodiversity and the principles of landscape ecology, terrace benches, bunds, and risers were planted with trees, shrubs, forage grasses, and crops, serving to generate a diverse array of plants, a semi-forested area, and to stabilize terrace bunds. Soft-ridged bench terrace made it possible to significantly reduce hazards arising from gravity erosion, and reduce the costs of individual bench construction and maintenance by 24.9% and 55.5% of the costs under traditional techniques, respectively. Such a construction allowed an enrichment and concentration of nutrients in the soils of terrace bunds, providing an ideal environment for a range of plants to grow and develop. The terrace riser could be planted with drought-resistant plants ranging from forage grasses to trees, and this riser vegetation would turn the exposed bunds and risers existing under traditional techniques into plant-covered belts, great green ribbons decorating farmland and contributing to the enhancement of the landscape biology. PMID- 16262057 TI - [Effects of irrigation and nitrogen fertilization on winter wheat yield under straw mulch]. AB - A field experiment was conducted on a Hongyou soil of Yangling to investigate the effects of irrigation and nitrogen fertilization on wheat yield under straw mulch. The results showed that under straw mulch, N fertilization contributed more to the increase of wheat yield than irrigation, because soil moisture condition was improved greatly. The interaction between irrigation and nitrogen fertilization was negative in no-mulch treatment, but positive under straw mulch. For dryland, more attention should be paid to the input of nutrients when straw mulch was applied. High moisture content in soil profile before sowing and sufficient N input were essential for good harvest when field was not mulched. Under straw mulch, the irrigation rate for a maximum yield was reduced, and the optimum time of irrigation was postponed. Wheat grain yield had no relation to the irrigation during jointing stage in both no-mulch and straw mulch treatments. PMID- 16262058 TI - [Relationships between red soil enzyme activity and fertility]. AB - Correlation and cluster analyses on the enzyme activities and chemical-biological properties of eight red soils showed that soil urease, invertase, phosphatase and catalase activities correlated significantly with soil organic carbon, total nitrogen and total phosphorous. Similar results of soil fertility evaluation were obtained by using soil enzyme activities and by using soil chemical-biological properties, indicating that soil enzyme activity could be used as an index of evaluating red soil fertility. The enzyme activities of fresh soil were generally greater than those of air-dried sample, and more closely correlated with soil fertility. PMID- 16262059 TI - [Dynamics of water retaining capacity and chlorophyll content of two-line hybrid rice during heading-grain filling stage and their relations with grain yield]. AB - The study with 25 two-line hybrid rice strains showed that their water retaining capacity was different with their combinations. On the whole, the water content in leaf and stem-sheath decreased during heading-grain filling stage, while that in panicle increased first and then decreased, with a peak at 7th day after heading. Stem-sheath was the main organ for water retaining. The correlation of panicle yield with both leaf and stem-sheath water content was positive at the 1st day after heading but became negative later, and that with whole stem-plant water content was positive after heading. The chlorophyll content of the two-line hybrids increased first and then decreased, with a peak at the 7th day after heading. Comparing with normal rice (male parent), the chlorophyll content of most two-line hybrids was lower, and the decreasing rate was faster than that of normal rice after heading. For two-line hybrid rice breeding, it should preferentially consider to select those hybrids which have a clearer dominance of plant water retaining capacity and a higher chlorophyll content at earlier and middle heading-grain filling stage, and an obvious dry matter transportation from leaf and stem-sheath at later heading-grain filling stage. For two-line hybrid rice planting, it should not only pay attention to the supply of water and nutrients, especially nitrogen at earlier and middle heading-grain filling stage, but also control nitrogen and water in time to prevent leaf green clinging and to promote dry matter transportation from leaf and stem-sheath to grain at later heading-grain filling stage. PMID- 16262060 TI - [Effects of shading at blossoming and boll-forming stages on cotton fiber quality]. AB - The study on the effects of different shading level at blossoming and boll forming stages on cotton fiber quality of Zhongmiansuo No.41 and Lumianyan No. 18 showed that with increasing shading, the maximum fiber length of cotton decreased, while the elongation period increased. The fiber length in 70% shading treatment was 1.01 mm shorter than that in 40% shading treatment. Without shading, the fiber reached its maximum length 25 days after anthesis, while in shading treatments, the fiber reached its maximum length 35 days after anthesis. Fiber gauge tenacity was also decreased with increasing shading. Comparing with the control, both 40% and 70% shading significantly decreased the fiber maturation and maturity. Two test cotton varieties presented the same change trend under shading condition. PMID- 16262061 TI - [Representativeness of Northeast China spring soybeans and their genetic diversity at SSR loci]. AB - A total of 283 accessions were selected from the total 3 226 Northeast China spring soybeans, which represented > 80% of the whole based on their qualitative and quantitative traits. The representative samples were analyzed by 61 SSR loci, and a total of 534 alleles were detected, ranging 2 - 16 alleles per locus, with an average of 8.75 alleles per locus. Among the accessions, the Simpson diversity index (SDI) for each locus ranged from 0.406 to 0.886, with a mean of 0.704, which was relatively lower since there were dominant alleles at most of loci in the representative samples. 35 accessions had specific alleles, which distributed among 29 loci. The differentiation coefficient was lower than 9.27% in 61 loci among the three provinces of Northeast China, which might be caused by lots of common alleles shared among these provinces. The genetic diversity in Jilin and Liaoning provinces was nearly equal, but was higher than that in Heilongjiang Province. The landraces of the representative samples at 61 loci had a higher genetic diversity than that of the released cultivars. The genetic diversity appeared within and between the provinces could be used to broaden the genetic base of modern cultivars, and also, the landraces were the major source for soybean breeding because of their high genetic diversity. PMID- 16262062 TI - [Physiological and ecological effects of inter- and mixed cropping rape with milk vetch]. AB - The investigation on the growth, yield and benefit of rape inter- and mixed cropped with milk vetch showed that comparing with sowing rape under zero tillage, the physiological and ecological characters of transplanted rape, including its height, leaf size, root diameter, opening degree and yield were obviously improved. Under zero tillage, mixed cropping had an obvious advantage than inter- and single cropping. After tillage, the yield of transplanted rape under mixed cropping was 11.9% more than that under single cropping with zero tillage. The benefit of transplanting rape under mixed cropping with tillage was the biggest, followed by single cropping rape with zero tillage. It could be concluded that the patterns of transplanting rape under mixed cropping after tillage and zero tillage had the best physiological and ecological effects, not only increasing rape yield and income, but also improving soil fertility. PMID- 16262063 TI - [Root activity and nitrogen assimilation of rice (Oryza sativa) under free-air CO2 enrichment]. AB - With Free-Air CO2 Enrichment(FACE) technique, this paper studied the root activity and amino acid(aa) synthesis of rice (Oryza sativa )at low N(LN, 150 kgN x hm(-2)) and normal N(NN, 250 kgN x hm(-2)) under ambient air and elevated atmospheric CO2 (Ambient + 200 micromol x mol(-1)). Under elevated CO2, the xylem exudates per hill changed little, while the xylem exudates per stem declined by 1.4% - 21.7% as the result of greater tiller numbers. At tillering and heading stages, elevated CO2 increased aa N/inorganic N in xylem exudates by 11.1% - 143.1%, but did not affect the aa concentration in xylem exudates and the total amount of aa in roots significantly. However, at 35 days after heading, the aa N/inorganic N ratio decreased by 38.1% (LN) and 29.2% (NN) under elevated CO2. FACE also declined the aa concentration in xylem exudates by 34.0% (LN) and 44.7% (NN), and the total amount of aa by 50.8% (LN) and 40.0% (NN), which meant the retarded capability of aa synthesis in roots. N amendment led to a decrease of aa N/inorganic N in xylem exudates by 19.5% (Ambient) and 36.8% (FACE) at heading stage, as the result of unaffected aa and increased inorganic N concentration. There existed a significantly antagonistical CO2 x N interaction on aa N/inorganic N at heading stage. PMID- 16262064 TI - [Effects of simulated acid rain and its acidified soil on soluble sugar and nitrogen contents of wheat seedlings]. AB - The study showed that the cation release of simulated rain caused soil acidification and base ions release. With the decrease of simulated acid rain pH from 5.6 to 2.5, the acid rain-leached soil pH decreased from 6.06 to 3.41, and its total amount of exchange base ions decreased from 56.5 to 41.1 mmol x kg(-1). Spraying simulated acid rain on the shoots of wheat seedlings planted on such acidified soils caused a rapid decrease in the soluble sugar and nitrogen contents of wheat seedlings, and reduced some of their physiological activities. The effect of spraying simulated acid rain on the soluble sugar, nitrogen, and chlorophyll contents and photosynthetic rate of wheat stems and leaves was larger than that of acidified soil, while the effect of the latter on the soluble sugar and nitrogen contents and the physiological activity of NR and GOGAT in root system of wheat seedlings was larger than that of the former. The intensive acid rain of pH < or = 3.0 and the corresponding acidified soil had an obvious harm to the growth and physiological activity of wheat seedlings. PMID- 16262066 TI - [Effects of alkali-stress on Aneurolepidium chinense and Helianthus annuus]. AB - Employing monocotyledon Aneurolepidium chinense and dicotyledon Helianthus annuus, the two species with high alkali-saline resistance as test materials, and stressing them with neutral, alkalic, and mixed salts, this paper studied the characteristics of various stresses and their interrelations, with relative growth rate (RGR) as the main strain index. The results showed that under the same concentration, alkalic salt had a stronger effect than neutral salt, and the RGR of A. chinense and H. annuus was decreased with increasing salt concentration under the same kind of salt stress and pH conditions. When the pH was higher, the RGR decreased more obviously. In addition, the proline and citric acid contents in test plants were increased with increasing stress. The transformation of proline in A. chinense was greater than that in H. annuus, while that of citric acid was in adverse. In a word, alkalic salt stress was different from neutral salt stress in stressing plants and in plant responses. Alkalic salt stress and neutral salt stress were actually two distinct kinds of stresses. The former was better called "alkali-stress", while "salt-stress" only meant neutral salt stress. The key difference between them was their different pH value. It was reasonable to consider the buffer capacity as the strength value of alkali stress, and the salinity as the strength value of salt-stress. An interactive effect between salt-stress and alkali-stress could be seen under mixed saline and alkali stress. PMID- 16262065 TI - [Effects of fungicide on temporal and spatial niches of Rhizoctonia solani]. AB - This paper studied the variation of the temporal and spatial niches of rice sheath blight-causing Rhizoctonia solani under spraying fungicide. The results showed that between fungicide treatments and control, the spatial niche breadth of Rhizoctonia solani was different, while its temporal niche breadth was similar. The spatial niche breadth of Rhizoctonia solani was 0.5240 when spraying fungicide at booting and full heading stages, 0.5742 at booting and milkfilling stages, and 0.8577 at tillering stage, while the control was 0.8563. Spraying fungicide had little effect on temporal niche breath. The spatial niche breath, percentage of diseased leaves and sheathes, index of disease, and control effect all suggested that spraying fungicide two times in rice growth period was better than spraying it one time. Spraying fungicide at booting and full heading stages and at booting stage and milkfilling stages had the best effects. Fungicide could not only narrow the spatial niche breadth, limit the spread of Rhizoctonia solani on top leaves which were important for the yield, but also narrow the temporal niche breadth, limit the spread of Rhizoctonia solani during the yield formation period of rice. But, spraying fungicide only changed the distribution of the fungus niches and narrowed the niches during rice yield formation period, with no changes in the whole niches of Rhizoctonia solani. PMID- 16262067 TI - [Effects of eutrophication on distribution and population density of Corbicula fluminea and Bellamya sp. in Chaohu Lake]. AB - The investigation on the distribution an d population density of C. fluminea and Bellamya sp. in Chaohu Lake during September 2001 and September 2002 showed that in the west region of the lake where was seriously eutrophic, the density and biomass of C. fluminea were 5.1 ind. x m(-2) and 17.87 g x m(-2) in 2001, and 8.8 ind. x m(-2) and 47.29 g x m(-2) in 2002, while those of Bellamya sp. were 13.3 ind. x m(-2) and 45.45 g x m(-2) in 2001, and 3.8 ind. x m(-2) and 12.56 g x m( 2) in 2002, respectively. In the east region of the lake where was eutrophic, the density and biomass of C. fluminea were 23.8 ind. x m(-2) and 67.86 g x m(-2) in 2001, and 29.2 ind. x m(-2) and 96.18 g x m(-2) in 2002, while those of Bellamya sp. were 10.1 ind. x m(-2) and 32.00 g x m(-2) in 2001, and 9.4 ind. x m(-2) and 31.21 g x m(-2) in 2002, respectively. The density and biomass of C. fluminea and Bellamya sp. were declined with increasing eutrophication. In hypertrophic region, C. fluminea and Bellamya sp. were absent. The density and biomass of the two species were obviously higher in littoral than in pelagic region. The distribution type of C. fluminea was core-model, while that of Bellamya sp. was random. The correlation between the density and biomass of C. fluminea and Bellamya sp. and water depth was not significant (P > 0.05). The biomass of Bellamya sp. was negatively correlated with water TN (P < 0.01), NO3-N (P < 0.05), TP(P < 0.01) and PO4-P (P < 0.05), while that of C. fluminea only had a significantly negative correlation with PO4-P(P < 0.05). Compared with 1981, there was fewer C. fluminea in the lake nowadays. The effects of other environmental factors on the population distribution and growth of C. fluminea and Bellamya sp. were also discussed. PMID- 16262068 TI - [Relationships between antioxidant activities and heat-resistant features of two Laminaria japonica strains]. AB - The study on the relationships between antioxidant activities and heat-resistant features of two Laminaria japonica gametophyte mutants Kelp 901(abbreviated as 901)and Rongcheng No. 1 (abbreviated as RC) under different temperature conditions showed that under normal temperature condition (10 degrees C), there were no positive relationships between the basal antioxidant activities and heat resistant features of 901 and RC, while under heat stress(18 degrees C), the reactive oxygen species (ROS) contents kept constant in 901, but accumulated rapidly in RC, suggesting that ROS played an essential role in oxidative stress. The enzyme activities in RC were more vulnerable to high temperature, and its antioxidant enzyme activities decreased more sharply under heat stress. Ascorbic acid (ASA), vitamin E, superoxide dismutase (SOD) and glutathione peroxidase (Gpx) might play important roles in kelp's heat-resistance. Chlorophyll a was sensitive to heat stress, and its 50% nocuity time could be used as an indicator to distinguish kelp strains with different heat-resistance. It was suggested that the antioxidant system's responses to heat stress instead of its basic activities accounted for the kelp's heat-resistance. PMID- 16262069 TI - [Factor analysis of Beijing north ring water system eutrophication]. AB - Eutrophication is caused by hydrological, physical or chemical factors such as temperature, illumination, or shapes of waters. It is very important to identify the main factors to understand and control the process of eutrophication. In this paper, Beijing north circle water system was divided into river subsystem and lake subsystem by cluster analysis. Factor analysis indicated that the main factors of river subsystem eutrophication were TP, TN and NH4-N, while those for lake subsystem were TN and NH4-N, which meant that in the study area, the main cause for eutrophication was the overload of nutrients. Stepwise regression was introduced to build forecasting model, and the choice results of independent variables in the regression model inferred that the type of eutrophication in river subsystem was P limited, while that of lake was N limited. Two main causes for nutrient load were found from water quantity and quality analysis of the study area, one was the ecological water demands be not able to satisfy from 1990 to 1998 except 1998, and the other was the increase of nutrient load coming from domestic waste and non-point pollutant, resulting from the rapid population increase and city expansion. The measures to control the eutrophication process were put forward. PMID- 16262070 TI - [Effects of atrazine and its degrader Exiguobaterium sp. BTAH1 on soil microbial community]. AB - The study showed that the application of atrazine stimulated soil microorganisms obviously. In comparing with control(without atrazine), the respiration intensity of soil applied with 50 mg atrazine kg(-1) soil increased greatly, the concentrations of soil NH4(+)-N and NO3(-)-N changed significantly, and the individuals of soil microbes, especially bacteria and fungi, also increased greatly. The application of strain BTAH1 could degrade 98% of applied atrazine within one week, and led to the decrease of soil respiration intensity. Under BTAH1 application, the individuals of actinomyces and fungi decreased, while those of bacteria did not, and the concentrations of soil NH4(+)-N and NO3(-)-N came back to the level of the control. ARDRA analysis on the 16s rDNA library of soil bacteria suggested that the application of atrazine could decrease the biodiversity of soil microorganisms, while applying BTAH1 could recover the biodiversity. PMID- 16262071 TI - [Effects of Bt toxin Cry1Ac on biochemical responses of Eisenia fetida in an artificial soil]. AB - Bt toxin Cry1Ac could be introduced into soil by pollen, root exudates, and plant residues. To investigate its toxic effects, the Bt toxin was added into an earthworm-cultured artificial soil, and the survival, growth rate, sperm count, protein content, and activities of acetylcholinesterase (AchE), catalase (CAT), glutathione S-transferase (GST) and cellulase in Eisenia fetida were determined. The results suggested that Bt toxin Cry1Ac had little effects on the biomass and physiological characteristics of E. fetida, with no acute and subchronic toxicity, and was safe for earthworm at field dose level. PMID- 16262072 TI - [Acute toxicological effects of excessive Cu and Zn-containing in pig manure on earthworm]. AB - Cu and Zn are popularly used as additives in animal feed in China, which could result in their excessive accumulation in manure to a toxic level, and thus, possibly threaten the environment when the manure is applied to farmland. In this paper, the acute and sub-acute lethal effects of pig manure with excessive Cu and Zn on earthworm were studied, and the results indicated that the concentration of Cu and Zn in pig manure had a significantly positive correlation with the mortality of earthworm, while a significantly negative correlation with earthworm growth rate. The individuals of earthworm had different tolerance to excessive Cu and Zn, with the threshold values causing death being 250 mg x kg(-1) and 400 mg x kg(-1) for Cu and Zn, respectively. The LD50 was 646.68 mg x kg(-1) for Cu, and 947.38 mg x kg(-1) for Zn. A strong synergistic effect was observed under the combined pollution of 250 mg x kg(-1) for Cu and 500 mg x kg(-1) for Zn, whereas antagonistic effect happened when 750 mg x kg(-1) for Cu was supplied, suggesting that the joint toxic effects of Cu and Zn on earthworm were closely correlated to their concentrations in pig manure. PMID- 16262074 TI - [Discussion on hydrologic scaling]. AB - Hydrologic scaling is the hotspot and frontier of today's hydrologic research, which includes three different meanings distinguished from each other, i.e., process scale, observation scale and modelling scale. As a characteristic of hydrologic scale, dominant process scale has been paid more attention. Hydrologic scaling includes the scaling of hydrologic models, parameters, state variables and inputs, which owns special research methods respectively. The difficulty of hydrologic scaling mainly comes from the organized complexity of hydrologic systems, the heterogeneity and variability in space and time, and the insufficiency of data. The ubiquitous self-similarity of natural river networks makes itself an important part of hydrologic scaling research. The combination of various techniques and theories is needed to motivate hydrologic scaling research. PMID- 16262073 TI - [Forest ecosystem service and its evaluation in China]. AB - Facing the relative lag of forest ecosystem service and estimation in China, this paper proposed to quickly carry out the research on the evaluation of forest ecosystem service. On the basis of the classification of forest ecosystem types in China, the service of artificial and semi-artificial forest ecosystems was investigated, which was divided into eight types, i.e., timber and other products, recreation and eco-tourism, water storage, C fixation and O2 release, nutrient cycling, air quality purifying, erosion control, and habitat provision. According to the assessment index system for global ecosystem service proposed by Costanza et al., a series of assessment index system suitable for Chinese forest ecosystem service was set up, by which, the total value of forest ecosystem service in China was estimated to be 30 601.20 x 10(8) yuan x yr(-1), including direct and indirect economic value about 1 920.23 x 10(8) and 28 680.97 x 10(8) yuan x yr(-1), respectively. The indirect value was as 14.94 times as the direct one. The research aimed to bring natural resources and environment factors into the account system of national economy quickly, and to realize the green GDP at last, which would be helpful to realize sustainable development and environment protection. PMID- 16262075 TI - [Nematodes as bioindicator of soil health: methods and applications]. AB - This paper summarized the advantages of using nematodes as soil health bioindicator. A review of commonly used indices and analyzing methods by means of nematodes was presented, with emphases on Maturity Index, as well as the indices and analyzing methods based on their trophic groups. Compared with other indices, Maturity Index can reflect the stressed degree of soil more sensitively, and the indices and analyzing methods based on their trophic groups can well reveal the state of soil health based on ecosystem functioning. The applications of nematodes as bioindicator in agriculture, forest, grassland, and other ecosystems were also outlined. In view of some practical limitations in the methods, prospective researches and applications were suggested, i.e., intensify the ecological studies to advance the understanding on the characteristics of soil nematodes, such as their life histories and feeding habits, consummate the indication system by integrating various nematode indices and analyzing methods, and extend the applications of soil nematodes in different ecosystems and in different regions at large scales. PMID- 16262076 TI - [Affecting factors of soil microorganism and root respiration]. AB - Soil respiration is an important part of the carbon cycle in terrestrial ecosystems, and its contribution to the global carbon budget has been the focus of wide concern. Researches on the affecting factors of soil microorganism and root respiration, the main components of soil respiration, contribute to the understanding of the role of soil respiration on global carbon cycle, and to the accurate evaluation of global carbon budget. This paper reviewed the direct and indirect affecting factors of soil microorganism and root respiration, including climate factors, soil properties, vegetation and litterfall, air CO2 concentration, and human activities. These affecting factors were interactive, and their contributions to soil microorganism and root respiration varied in temporal and spatial scales. The study on natural and anthropogenic factors of soil microorganism and root respiration was emphasized, and some related research tasks in the future were also proposed. PMID- 16262077 TI - [Research advances on eco-chemical behaviors and toxicological effects of cadmium in root-soil interface]. AB - Many active substances such as organic acids and enzymes excreted by living plant roots could induce a great difference of Eh and pH values between root-soil interface and non-rhizosphere soil, forming a special root-soil interface miniature environment. As a mini-type ecological area with most frequent exchanges of substances, root-soil interface plays a crucial role in their absorption, transformation, migration and eco-toxicological effects. In this paper, the eco-chemical behaviors of Cd in root-soil interface affected by the change of pH, Eh and root secretion, and its eco-toxicological effects on microorganisms and enzymes in root-soil interface were reviewed, based on the related research advances in recent decade. The shortages in relevant fields were pointed out, and the scientific problems to be researched in the future were suggested. PMID- 16262078 TI - [Advances in research and application of water-treatment flocculants]. AB - Water-treatment flocculants are being developed from traditional to inorganic, from inorganic to organic, from natural organic to synthetic organic, from synthetic organic to modified natural organic, from inorganic or organic to inorganic-organic, and from chemical to biological with ecological safety. In order to promote the rapid development and practical application of water treatment flocculants, the research progress and application situation of flocculants for water treatment at home and abroad were reviewed. In particular, the characteristics of two important flocculant types, chemical and microbiological flocculants, as well as their application processes in water treatment were summarized, their development orientations were analyzed and evaluated, and the future research directions aimed to overcome the current shortages in developing and applying flocculants in China were put forward. PMID- 16262079 TI - [Recycling rate of N and P through a feeding-composting cycle and their recoveries in agro-ecosystems]. AB - A five-year experiment indicated that the average loss rate of N and P in harvested products through a feeding-composting cycle was about 50% and 15%, respectively. Under high yield condition, the amount of recycled N and P from 80% harvested products and through a feeding-composting cycle in farming systems was about 37 - 51 and 8 -14 kg x hm(-2), equivalent to 25% - 34% of N and 32% - 56% of P from chemical fertilizers applied each year to the systems. The apparent recoveries of N and P in organic manure increased with the prolongation of fertilization, indicating a synergetic residual effect existed, and was 61% and 39% in average in the five-year experiment. The use of nutrients recycled in the farming systems could not only improve soil fertility, but also increase the recoveries of nutrients and reduce the use of chemical fertilizers. PMID- 16262080 TI - [Biodegradation of di-n-butyl phthalate by mangrove microorganism Rhodococcus ruber 1K]. AB - A di-n-butyl phthalate (DBP)degrading bacterium Rhodococcus ruber was isolated from mangrove soil, and its degrading characteristics were studied. The results showed that the bacterium could grow well on the substrate with DBP as the sole source of carbon and energy, and the DBP of 50 mg x L(-1) could be completely degraded after 48 h. Under aerobic condition, the tentative pathway proposed for DBP degradation was through monoester initially, then phthalic acid, and finally CO2 and H2O. PMID- 16262082 TI - [Extraction of active substance from Chlorella vulgaris cells and its physiological effects on Saccharomyces cerevisiae]. AB - The study showed that freezing-thawing method could have a higher cellwall breakage rate, and a light injury to the active substance, which could not only increase the cell fission and growth, delay the death of Saccharomycete cells, but also enhance CO2 production rate and quicken fermentation process when the cells were exposed to the medium with 0.5% active substance. PMID- 16262081 TI - [Characteristics of Yunnan Province rice landrace core collections under phosphorus deficiency]. AB - Soil phosphorus (P) deficiency is a major yield-limiting factor in rice production. Employing 526 rice landrace accessions from 5 rice-planting regions and 16 prefectures in Yunnan Province, this paper studied their P-deficiency tolerance characteristics by comparing the relative plant dry weight under acid red soil condition. The results showed that Northwest Yunnan cold highland japonica rice-planting region was most abundant in P-deficiency tolerance core collection of indica, and there was no significant difference in japonica among five ecological zones of rice landraces. Nujiang, Lijiang, Dehong, Wenshan, Xishuangbana and Yuxi were most abundant in P-deficiency tolerance core collection of indica, and Lijiang, Wenshan and Chuxiong were most abundant in P deficiency tolerance core collection of japonica. Indica had a higher P deficiency tolerance than japonica. The P-deficiency tolerance of rice was significantly related with its genetic diversity, rice breeding and acid red soils. PMID- 16262084 TI - "Growing up" with JGN: new ideas are waiting in your break room. PMID- 16262083 TI - [Application effect of most probable number (MPN) method in photosynthetic bacteria counting]. AB - The application of most probable number (MPN) method in counting the numbers of photosynthetic bacteria (PSB) showed that this method was suitable for PSB counting under illumination and anaerobic condition, which remedied the shortages of other methods. With this method, the sensitivity and accuracy of the counting could be enhanced when more replicated tubes and lower dilution times for fewer bacteria samples were applied. When PSB were not dominant in soil and water, MPN method could be used instead of other methods. PSB agent could only survive 6 approximately 9 days, and not be dominant in the environment in a long-term. Its application time and oxygen level affect the numbers of PSB significantly, which were the largest in a short application time and under anaerobic condition. PMID- 16262085 TI - Creating a multisensory environment for dementia: the goals of a Snoezelen room. AB - The MR provides a means for a variety of sensory experiences that can increase the quality of life for an individual with dementia. It is an example of an innovative approach to meeting the social and recreational needs of individuals with dementia. In addition, the MR enables staff and family members to communicate and build relationships with residents on a new and different level. PMID- 16262086 TI - The rural community care gerontologic nurse entrepreneur: role development strategies. AB - Rural elderly individuals are an underserved population with limited access to health care. There is an increasing need for independent community care nurses to provide assistance to home-based elderly individuals with chronic illnesses to prevent unnecessary medical and placement decisions and, thus, allow them to maintain independence and quality of life. This article describes the rural setting and why community care nurses are needed, and explores strategies for implementing the role of the independent nurse entrepreneur in caring for community-based elderly individuals in rural settings. PMID- 16262087 TI - Perceived health and life satisfaction among older adults: the effects of worry and personal variables. AB - The purpose of this study was to describe how worry affects two desired outcomes of nursing care for older adults-perceived health and life satisfaction. A convenience sample of 100 individuals (ages 65 to 97) living in the community responded to the questionnaire. Despite several chronic diseases, 89% of the participants perceived themselves as "well." Descriptive statistics revealed relatively high life satisfaction. Data from the Perceived Health Scale indicated the participants were functioning near their normal capacity, and with few negative emotions and physical sensations. Worry was also minimal. Nevertheless, using multiple regression, worry contributed to lower life satisfaction, negative emotions, physical discomforts, and decreased functional ability. Because the reported worries were primarily related to health and functioning, assessment of worry is significant when planning nursing care for older adults. PMID- 16262088 TI - Wearing and using personal emergency respone system buttons. AB - Frail older adults living alone are at risk for falling and not being found. A personal emergency response system (PERS) is a technological adjunct to home care for such individuals, but little is known of their experiences with the PERS. The experiences of seven frail women (ages 83 to 96) who lived alone were studied using a descriptive phenomenological method. The key phenomenon was temporizing about the PERS button, and the two component phenomena were deciding when to wear it and deciding whether to use it. Findings were contrasted to the existing literature about wearing and using the PERS, which has been focused on compliance. The variations in the experiences of the women in this small sample warrant further research and individualized interventions to enhance consistent use of the PERS by frail older adults. PMID- 16262089 TI - Prostate cancer: appraisal, coping, and health status. AB - The purpose of this study was to identify how cognitive appraisal and types of coping affect the health status of men with prostate cancer. Lazarus and Folkman's model of stress and coping guided this correlational, cross-sectional study. The convenience sample was composed of 131 men with prostate cancer who completed the Cognitive Appraisal of Health Scale, the Ways of Coping Checklist, and the Short-Form Health Survey using mailed questionnaires. Participants who appraised more harm or loss experienced worse physical and mental health. When participants perceived their diagnosis as posing more harm or loss or a greater threat, they were more likely to use emotion-focused coping. When the diagnosis was perceived as a challenge, men were more likely to use more problem-focused coping. The findings of this study enable health care providers to be more attentive to the psychosocial needs of prostate cancer patients. PMID- 16262090 TI - Spirituality and health: perceptions of older women in a rural senior high rise. AB - The purpose of this study was to examine the essence of spirituality and health in older women living in a rural senior high-rise apartment building. The researchers used a phenomenological research design and interviewed 10 women 65 years of age and older. Data analysis using Colaizzi's (1978) method for analysis revealed three themes-health is functional and provides a sense of wholeness; the relationship with God or a higher power is a personal one; and death is a part of life. The three themes create an essential structure, which influences the way the participants live. These findings provide information for nurses on the spiritual needs of older women and how the experiences of living in a high rise affect spirituality and health. PMID- 16262091 TI - The passenger. PMID- 16262092 TI - Intervention, in its place. PMID- 16262093 TI - Pre-term birth: the risk factors. PMID- 16262094 TI - An early warning system for pre-eclampsia. PMID- 16262095 TI - Why mothers die. PMID- 16262096 TI - Midwifery basics: complications in pregnancy (1). Antepartum haemorrhage. PMID- 16262097 TI - The language of the complicated. PMID- 16262098 TI - Managing cord prolapse at a home birth. PMID- 16262099 TI - Clinical risk management (2): Understanding the causes of failure. PMID- 16262101 TI - More than a cuddle: skin-to-skin contact is key. PMID- 16262100 TI - When a simple 'yes' or 'no' is not enough. PMID- 16262102 TI - An education in training. PMID- 16262103 TI - Can psychiatrists prevent suicide? Yes, in collaboration. PMID- 16262104 TI - Can psychiatry prevent suicide? Not yet! PMID- 16262105 TI - Comorbid depression among untreated illicit opiate users: results from a multisite Canadian study. AB - OBJECTIVES: This study aimed to describe patterns of major depression (MDD) in a cohort of untreated illicit opiate users recruited from 5 Canadian urban centres, identify sociodemographic characteristics of opiate users that predict MDD, and determine whether opiate users suffering from depression exhibit different drug use patterns than do participants without depression. METHOD: Baseline data were collected from 679 untreated opiate users in Vancouver, Edmonton, Toronto, Montreal, and Quebec City. Using the Composite International Diagnostic Interview Short Form for Major Depression, we assessed sociodemographics, drug use, health status, health service use, and depression. We examined depression rates across study sites; logistic regression analyses predicted MDD from demographic information and city. Chi-square analyses were used to compare injection drug use and cocaine or crack use among participants with and without depression. RESULTS: Almost one-half (49.3%) of the sample met the cut-off score for MDD. Being female, white, and living outside Vancouver independently predicted MDD. Opiate users suffering from depression were more likely than users without depression to share injection equipment and paraphernalia and were also more likely to use cocaine (Ps < 0.05). CONCLUSIONS: Comorbid depression is common among untreated opiate users across Canada; targeted interventions are needed for this population. PMID- 16262106 TI - Obsessive-compulsive symptoms and positive, negative, and depressive symptoms in patients with recent-onset schizophrenic disorders. AB - OBJECTIVES: To study the relation between obsessive-compulsive symptoms (OCS) and positive, negative, and depressive symptoms in patients with recent-onset schizophrenic disorders. METHODS: We undertook a prospective study of 113 consecutively hospitalized patients with recent-onset schizophrenia or related disorders diagnosed according to DSM-IV criteria. We compared 3 subgroups: one without comorbid OCS, one with OCS not fulfilling DSM-IV criteria for obsessive compulsive disorder (OCD), and one with comorbid OCD diagnosed according to DSM IV criteria. We assessed OCS severity at admission and 6 weeks thereafter with the Yale-Brown Obsessive Compulsive Scale (Y-BOCS). The Positive and Negative Syndrome Scale (PANSS) and the Montgomery-Asberg Depression Rating Scale (MADRS) were independently administered. RESULTS: At admission, patients with schizophrenic disorders and OCD had higher mean MADRS scores than both other groups; patients with OCS not fulfilling DSM-IV criteria for OCD had lower mean PANSS negative subscale scores than both other groups. After 6 weeks, there were no significant between-group differences, and OCS severity remained constant. CONCLUSIONS: Acute patients with recent-onset schizophrenia and OCD have more severe depressive symptoms but do differ in negative symptoms, compared with patients without comorbid OCD. Mild OCS may be related to less severe negative symptoms. During regular inpatient treatment, OCS severity remains constant PMID- 16262107 TI - Facial expression and sex recognition in schizophrenia and depression. AB - BACKGROUND: Impaired facial expression recognition in schizophrenia patients contributes to abnormal social functioning and may predict functional outcome in these patients. Facial expression processing involves individual neural networks that have been shown to malfunction in schizophrenia. Whether these patients have a selective deficit in facial expression recognition or a more global impairment in face processing remains controversial. OBJECTIVE: To investigate whether patients with schizophrenia exhibit a selective impairment in facial emotional expression recognition, compared with patients with major depression and healthy control subjects. METHODS: We studied performance in facial expression recognition and facial sex recognition paradigms, using original morphed faces, in a population with schizophrenia (n=29) and compared their scores with those of depression patients (n=20) and control subjects (n=20). RESULTS: Schizophrenia patients achieved lower scores than both other groups in the expression recognition task, particularly in fear and disgust recognition. Sex recognition was unimpaired. CONCLUSION: Facial expression recognition is impaired in schizophrenia, whereas sex recognition is preserved, which highly suggests an abnormal processing of changeable facial features in this disease. A dysfunction of the top-down retrograde modulation coming from limbic and paralimbic structures on visual areas is hypothesized. PMID- 16262108 TI - Informed consent and adolescents. AB - OBJECTIVE: To explore the doctrine of informed consent and the development of capacity in adolescents with psychiatric problems to help clinicians better reflect on the relevant ethical issues. METHOD: We discuss the relevant literature and explore the role of psychiatric impairment in adolescents' ability to consent. RESULTS: In common law, there is no minimum age at which individuals are able to consent to medical treatment and no age below which they are unable to consent. Adolescents' right to self-determination is based on their ability to understand and appreciate the information relevant to the medical decision and on their ability to consent voluntarily and freely. There is a consensus in the literature that, around age 14 years, adolescents have the cognitive ability to understand information necessary for consent. However, there are limited empirical data regarding adolescents' ability to appreciate the information and to make a voluntary decision. CONCLUSION: Clinicians need to involve adolescents in the consent process to the extent possible and assess the elements of capacity to consent to treatment on an individual case basis, recognizing that capacity may evolve as adolescents' cognitive capacities and values mature. PMID- 16262109 TI - Psychiatric training in rural and remote areas: increasing skills and building partnerships. PMID- 16262110 TI - Tardive dyskinesia in the era of typical and atypical antipsychotics. Part 1: pathophysiology and mechanisms of induction. AB - OBJECTIVE: Tardive dyskinesia (TD) is the principal adverse effect of long-term treatment with conventional antipsychotic agents. Several mechanisms may exist for this phenomenon. Mechanisms for the lower incidence of TD with atypical antipsychotics also remain to be fully understood. We undertook to explore and better understand these mechanisms. METHODS: We conducted a comprehensive review of TD pathophysiology literature from January 1, 1965, to January 31, 2004, using the terms tardive dyskinesia, neuroleptics, antipsychotics, pathophysiology, and mechanisms. Additional articles were obtained by searching the bibliographies of relevant references. Articles were considered if they contributed to the current understanding of the pathophysiology of TD. RESULTS: Current TD vulnerability models include genetic vulnerability, disease-related vulnerability, and decreased functional reserve. Mechanisms of TD induction include prolonged blockade of postsynaptic dopamine receptors, postsynaptic dopamine hypersensitivity, damage to striatal GABA interneurons, and damage of striatal cholinergic interneurons. Atypical antipsychotics may cause less TD because they have less impact on the basal ganglia and are less likely to cause postsynaptic dopamine hypersensitivity. CONCLUSION: Although the ultimate model for TD is not yet understood, it is plausible that several of these vulnerabilities and mechanisms act together to produce TD. The lower incidence of TD with atypical antipsychotics has helped to elucidate the,mechanisms of TD. PMID- 16262111 TI - Motivational interviewing and clinical psychiatry. AB - OBJECTIVES: Our objectives were as follows: (1) to survey the literature on motivational interviewing (MI), "a client-centered yet directive method for enhancing intrinsic motivation to change by exploring and resolving client ambivalence" and a well-established method of brief intervention, especially in the field of addictions treatment; (2) to review hypotheses about its mode of action; and (3) to discuss its possible impact on clinical psychiatry, in particular, on teaching communications skills. METHOD: Literature reviews and metaanalyses of numerous clinical trials of MI for addictions treatment have already been published and are briefly summarized. So far, no literature survey exists for MI applied to psychiatric patients. This review is limited to a synthesis of the articles relevant to psychiatry and to comments based on our team's experiences with MI. RESULTS: There is no evidence that MI achieves better results than other established techniques for treating addictions; it may simply work faster. The explanation for the method's rapid effectiveness remains speculative. Outcomes concerning the application of MI to psychiatric patients, although preliminary, are promising. Methods of assessing the integrity of MI treatment are more developed than in most psychotherapies, which permits the learning progress of trainees to be measured. CONCLUSIONS: MI offers a complement to usual psychiatric procedures. It may be worthwhile to teach it, not only for addictions but also for other broad treatment issues, such as enhancing patients' medication compliance and professionals' communication skills. Questions remain concerning MI's feasibility in psychiatry settings. PMID- 16262112 TI - Atypical antipsychotics in psychiatric practice: practical implications for clinical monitoring. AB - OBJECTIVES: To provide practical recommendations for monitoring patients both before and during treatment with atypical antipsychotics, to assist clinicians in implementing preventative measures against diabetes, and to establish baselines according to which clinicians should initiate diabetes treatment. METHOD: A working group of Canadian specialists in psychiatry and endocrinology reviewed peer-reviewed clinical studies published in this area and other relevant papers and abstracts. RESULTS: The reviewed studies further confirm that atypical antipsychotic medications are the most effective components in the medical management of many psychotic conditions; they also further emphasize the need to more stringently monitor and recognize diabetes risk factors inherent in these patients. Recommendations are based on a review of the available data, on expert opinion and consensus, and on current Canadian guidelines for the treatment of schizophrenia and management of diabetes. CONCLUSIONS: Patients with psychiatric disorders, most particularly schizophrenia and mood disorders, have an increased risk for type 2 diabetes and should be screened frequently, especially when other risk factors are present. The resulting recommendations offer practical steps for effectively screening patients prior to and during treatment with atypical antipsychotics. They include (1) how to conduct an initial baseline assessment, (2) when and how to monitor blood glucose and lipid levels, and (3) how to educate patients regarding such lifestyle issues as nutrition, exercise, and diet. PMID- 16262113 TI - Re: Diogenes syndrome in a pair of siblings. PMID- 16262114 TI - Ziprasidone-induced tardive dyskinesia. PMID- 16262115 TI - Re: Evaluation of a children's temper-taming program. PMID- 16262116 TI - Canadian depression prevalence. PMID- 16262117 TI - [Tattoo pigments contains toxic compounds, but legislators do not pay attention]. PMID- 16262118 TI - [Good practice in occupational health care]. PMID- 16262119 TI - [Head lice and crab lice]. PMID- 16262120 TI - [Degradation products of endothelial basement membranes: their role as tumor suppressor]. PMID- 16262121 TI - [Bradycardia due to hyperkalemia]. PMID- 16262122 TI - [Care of tonsillitis]. PMID- 16262123 TI - [Update on current care guidelines. Care of Crohn's Disease]. PMID- 16262124 TI - [Cardiac arrhythmia and hairless patient]. PMID- 16262125 TI - [Pacemaker cord making loops]. PMID- 16262126 TI - [Evidence based medicine, paradigm and values]. PMID- 16262127 TI - [Alone]. PMID- 16262129 TI - [Gynecologic examination of young girls are a multidisciplinary challenge]. PMID- 16262128 TI - [Child and adolescent gynecology, what is beyond contraception]. PMID- 16262130 TI - [From girl to woman, normal development during adolescent and most common abnormalities]. PMID- 16262131 TI - [Lack of menstruation and abnormal bleeding]. PMID- 16262132 TI - [Repeated abdominal pain of teenage girls, gynecological or other problem?]. PMID- 16262133 TI - [How obesity of girls during childhood and adolescent reflects to reproductive ability]. PMID- 16262134 TI - [Gynecologic problems due to cancer treatment during childhood and adolescence]. PMID- 16262135 TI - Mississippi's tiniest babies do well. PMID- 16262136 TI - Lessons of the heart. PMID- 16262137 TI - New beginnlngs. PMID- 16262138 TI - An interview with UMC Vice Chancellor Dan W. Jones, MD. Interview by Philip T Merideth. PMID- 16262139 TI - The birthing of UMC. PMID- 16262140 TI - Images in Mississippi medicine. J. Robert Snavely, MD (1913-1964), UMC'S first chairman of Department of Medicine. PMID- 16262141 TI - [Health risks of occupational exposure to anticancer (antineoplastic) drugs in health care workers]. AB - In Europe and America, many health care workers have concerns about the risks to their health of handling anticancer drugs. The findings of the risks were reported in the late 1970's, and guidelines for the safe handling of anticancer drugs were established in the 1980's. The conditions of wearing personal protective equipment and of the working environment have improved dramatically as a result of introduction of the guidelines. Furthermore, researches and studies into the health effects of occupational exposure to anticancer drugs have been pursued actively. In Japan, the society of hospital pharmacists established guidelines for the safe handling of anticancer drugs in 1991. Since then, mainly nurses have been concerned about the safe handling of anticancer drugs, but in the medical setting, the present situation surrounding the safe handling of anticancer drugs has hardly changed. In the industrial hygiene field, the safe handling and the occupational exposure to anticancer drugs have been seldom reported and researched. The actual potential hazards to occupational exposure of anticancer drugs have not yet been determined. Nevertheless, the reduction of occupational exposure to anticancer drugs in health care workers has been an important challenge for the industrial hygiene field. In Japan, we need to promote action to spread the use of the appropriate personal protective equipment and the appropriate working environment. We also have to rethink the safe handling of anticancer drugs. We hope that the Japanese government will establish an effective authorized guideline as has been done in Europe and America. PMID- 16262142 TI - [Consciousness survey regarding genetic diagnosis of glutathione S-transferase M1 (GSTM1) and aldehyde dehydrogenase 2 (ALDH2) polymorphism]. AB - A Consciousness survey regarding genetic diagnosis of GSTM1 and ALDH2 was performed to evaluate the potential use of such a diagnosis in supporting those wanting to stop smoking and decrease alcohol intake. A questionnaire was given to 1,654 employees (male: 1,225, female: 429) who worked at an LSI manufacturing factory, and 1,434/1,654 (86.7%) responded to the survey. The number of respondents who replied that they "wanted to know the results of the genetic diagnosis of GSTM1 and ALDH2" were 731/1,401 (52.2%) and 812/1,434 (56.6%), respectively while the numbers of respondents who replied that they "did not want to know the results" were 138/1,401 (9.9%) and 103/1,434 (7.2%), respectively. The main reasons given for wanting to know the results of the genetic diagnosis of their enzymes reflected the respondents' awareness of their genetic susceptibility. These reasons included a desire to know the effects of tobacco smoke, to prevent diseases in the future, to know the effects of passive smoking or to know their tolerance for alcohol. On the other hand, the main reason for not wanting to know the genetic results that the respondents had no intention of stopping smoking and heavy drinking, or that they would be unable to stop even if they knew the results of the genetic diagnosis. Multiple regression analysis showed that the number of respondents who "wanted to know the results of the genetic diagnosis" was significantly higher among those respondents who are current smokers (male: OR = 1.66 95%CI 1.29-2.14, female: OR = 2.33 95%CI 1.37 3.98), those who understood the relationship between smoking and lung cancer (male: OR = 1.81 95%CI 1.25-2.63, female: OR = 2.77 95%CI 1.42-5.40) and those who with a high CAGE test score (male: OR = 1.96 95%CI 1.42-2.68, female: OR = 2.52 95%CI 1.07-5.94). The results of this survey suggest that genetic diagnosis of GSTM1 and ALDH2 polymorphism may be useful in supporting those who want to stop smoking and decrease their alcohol intake. PMID- 16262144 TI - [Status of noise in small-scale factories having press machines and hearing loss in workers]. AB - A study on noise and hearing loss was conducted in 36 small-scale factories where press machinery is actively used. Noise levels in working environments were measured in 34 factories. Of those measured, two (6%), eight (24%) and 24 (71%) factories were classified into control classes I, II and III, respectively, according to guidelines established for the prevention of noise-related disorders. Furthermore, personal exposure levels to noise were measured for 23 workers. Twenty-one (91%) of those workers measured were exposed to 85 dB(A) or higher, (the occupational exposure limit for an 8-hour exposure period). Maximum exposure levels were found to be at 102 dB (A). Hearing tests were then conducted on 97 male workers at those 36 factory sites. Twenty (21%), 30 (31%) and 10 (10%) of the workers measured were classified into a "precursory symptom group", a "slight hearing loss group" and a "medium to serious hearing loss group", respectively, based on the aforementioned guidelines. It was found that as workers' ages increased, the percentage of workers having some degree of hearing loss increased. Specifically, it was found that there was some level of hearing loss for 93% of those studied aged in their 50's, and up to 100% in those aged in their 60's. PMID- 16262143 TI - [Effect of work and organizational characteristics on workers in call centers: longitudinal study in an information service company]. AB - To investigate the effect of work and organizational characteristics on workers' health status, with job dissatisfaction and intentions to leave as "organizational health", we conducted a longitudinal study using a questionnaire survey in call centers of an information service company from July to August in 2001 and 2002. The response rates were 96.2% and 92.0%, respectively. For the statistical analysis, the completed data of 296 technical support staff, which was more than 80% of the data, was used. We identified seven subscales composed of 29 items of work and organizational characteristics as scales of "organizational characteristics" and "work and workplace characteristics". The results of hierarchical multiple regression analysis showed quantitative and qualitative job-overload influenced psychological health status and poor supervisor support influenced all outcome variables. Moreover, "organizational characteristics" influenced cumulative fatigue and job dissatisfaction, showing an indirect effect with poor supervisor support and coworker support. This study suggests that measures of work and organizational characteristics are useful interventions for "organizational health". PMID- 16262145 TI - [Road to implementation in the surgical specialities]. PMID- 16262146 TI - [Effect and measure against introduction of diagnosis procedure complex]. PMID- 16262147 TI - [Effect of DPC based payment and its future]. AB - Since 2003, the DPC based bundle type payment scheme has been applied for the acute care hospitals, including the special function hospitals. The DPC scientific committee under the Central Social Insurance Medical Council has examined the change in hospital services after the introduction of DPC based payment. It has been observed the reduction of ALOS, resource uses such as laboratory test radiological examination, drugs and supplies. The under treatment due to strong incentive for cost containment has not been observed up to now. PMID- 16262148 TI - [Role of the medical institution in comprehensive medical care]. AB - The diagnosis procedure combination (DPC) of Japan started in April 2003. It is a unique "per day" payment system that evolved from diagnosis-related group and prospective payment system, which was a "per case" system. After a trial of two years, various undesirable problems of DPC have been revealed. A review of those problems in major diagnostic classifications (16) is in progress. The main problems are: 1) the medical institution-specific coefficient; 2) up-coding; and 3) consideration for expensive medical equipment such as during cardiac catheterization. The inclusive payment system is described and the problems of DPC are discussed. PMID- 16262149 TI - [Influence of DPC on cardiovascular surgery]. AB - After a new medical expense system "DPC (Diagnosis Procedure Combination)" had been introduced in 2003, the change in the medical treatment fee was evaluated according to each cardiovascular surgical field. Application of the DPC is limited to hospitalization fee in the general ward by a fixed daily amount The former payment system remains for operation, anesthesia, and some invasive examinations such as cardiac catheterization, therefore, the influence of the DPC introduction on cardiovascular surgery was minimum. A lot of problems are still left, however, for the classification of diagnosis, especially in a congenital heart disease, and the fee setting of intensive care. It is required also to reconsider serious cases, advanced complications and methods of life support. In the field of cardiovacular surgery, the treatment is so risky and so expensive, that more detailed analysis should be necessary by establishment of data base system. PMID- 16262150 TI - [Effects and measures of the inclusion payment system with disease procedure combination in a respiratory surgical division]. AB - There are advantages for hospital managers, payment associations, and politic side with the intrduction of the inclusion payment system with disease procedure combination (DPC). However, what will be the effects on patients and medical personnel? The average length of hospitalization of DPC cases for pulmonary malignant tumor in DPC in hospitals has been shortened. However, hospital stays have been becoming shorter in even in hospitals not subject to DPC, and it appears that there is little effect of DPC on this. It is difficult to confirm the effects of DPC introduction on changes in the quantitative and qualitative use of medical supplies and agents. Less thorough examinations upon admission are one of the most obvious results of DPC. However, preoperative examinations are performed in outpatient departments, and postoperative examinations have not been very much affected. DPC is not only useful as a tool for analyzing hospital management, but also as a tool to analyze the components of medical treatment. We can use DPC to improve medical quality. PMID- 16262151 TI - [Influence and provision for comprehensive insurance system--digestive surgery]. AB - The diagnostic procedure combination (DPC) was introduced in 82 institutions from April 2003. DPC is similar to the diagnosis-related group and prospective payment system, which is widely used in the USA. The payment of DPC is calculated by multiplying the cost/day in each DPC, hospital stay in days, and index of each hospital, which is determined by several variables including the mean hospital stay in the previous year. After the introduction of DPC, clinical and diagnostic examinations, and cancer chemotherapy were shifted from the inpatient to outpatient setting in our institution, as well as in most of the other institutions reported by the Ministry of Health, Labor and Welfare. Because of the examinations in the outpatient clinic, the preoperative hospital stay was shortened. To validate the safety and effectiveness of cancer chemotherapy in the outpatient clinic, a central system was established in our institution by unifying the protocols and limiting the maximum administered doses by computer. After introduction of DPC, the average hospital stay was shortened in most institutions including ours. In spite of the satisfaction of the patients surveyed, the benefits of DPC should be confirmed based on the final outcome in terms of clinical therapeutic results. PMID- 16262152 TI - [Effects and measures of the flat payment system based on the diagnosis procedure combination system in pediatric surgery]. AB - The current status of the flat payment system based on the diagnosis procedure combination (DPC) system was examined in pediatric surgery. Many important diseases especially in neonatal surgery are not listed for the DPC system due to either the small number of cases or variations in hospital stay or cost In our university hospital, however, the DPC system was applied to 286 (90.8%) of 315 admissions. Total scores for the admissions were slightly higher (103.5%) in the DPC system compared with the fee-for-service system. Scores for inguinal hernia by day surgery and one-night stay were also slightly higher (102.2%) in the DPC system. Future measures for the DPC system include the provision that most preoperative evaluations should be performed in outpatient clinics and further elective surgery at a separate admission is recommended for patients with benign disease or in good condition. In cases of surgery for emergencies or malignancies, too many examinations should be avoided. Postoperative treatment using a clinical path without complications should be performed to achieve shortening of hospital stays and reduced cost. Problems in the current DPC system are complicated and inadequate classification, probably due to the unique and cost-unbeneficial nature of pediatric surgery. To establish a better medical fee system, further efforts to improve the DPC system should be continued. PMID- 16262153 TI - [Current status and future perspectives of living kidney transplantation from ABO incompatible or spousal donors]. PMID- 16262154 TI - [What can we learn from a case of medical malpractice?]. PMID- 16262155 TI - [Evidence based antiviral therapy for chronic hepatitis B]. PMID- 16262156 TI - [Molecular mechanisms of circadian clock: how digestive organs tick?]. PMID- 16262157 TI - [Circadian rhythm in cancer: target for cancer?]. PMID- 16262158 TI - [A case of multiple gastric carcinoid without type A gastritis, but having hypergastrinemia]. PMID- 16262159 TI - [A case of adult's still disease complicated with cytomegalovirus infection and hemorrhagic rectal ulcer]. PMID- 16262160 TI - [Simultaneous mucosa-associated lymphoid tissue lymphoma of the stomach, jejunum, and colon]. PMID- 16262161 TI - [A case of gastric outlet obstruction by gallstone (Bouveret's syndrome) treated by EHL]. PMID- 16262162 TI - [A case of a small intrahepatic cholangiocarcinoma treated with microwave coagulation therapy]. PMID- 16262163 TI - [Two cases of enteropathy type T cell lymphoma]. PMID- 16262164 TI - [A case of gangrenous ischemic colitis complicated with sigmoid-ileal fistula]. PMID- 16262165 TI - [Case report: cystic metastasis in liver fifteen years after surgical operation of malignant melanoma]. PMID- 16262166 TI - [A case of sclerosing cholangitis trigered by 5-FU/leucovorine combination therapy and rapidly deteriorated]. PMID- 16262167 TI - [A case of pancreatic pleural effusion demonstrated on MRCP and successfully treated with endoscopic pancreatic drainage]. PMID- 16262168 TI - Co-creation theory. AB - The Design Council is working with healthcare staff and users to reconfigure services for the 21st century. This 'co-creation approach' has focused on diabetes and motivating older people to stay healthy. PMID- 16262169 TI - Culture matters. AB - Zebaa Arif reflects on changes during her career as a mental health nurse in relation to cultural care issues: Cultural awareness is becoming embedded in patient care. All aspects of care are influenced by cultural beliefs and should form part of assessment. Leadership is essential in influencing cultural care, as is organisational commitment. PMID- 16262170 TI - Freedom from racism. AB - Commission for Racial Equality chair Trevor Phillips has criticised the NHS for being racist. Since then the health service has introduced various initiatives to tackle the problem - but are they working? PMID- 16262171 TI - When rites are wrong. AB - 140 million girls and women worldwide have undergone some form of female genital mutilation (FGM). The African Women's Clinic at University College London Hospitals NHS Trust is one of eight UK centres providing care and support for women experiencing problems as a result of FGM. FGM is most commonly performed in Africa, the Middle East and Asia. FGM is illegal in the UK. FGM can cause recurrent urinary tract infection, painful sexual intercourse and menstruation, and difficulties with childbirth. A ten-minute procedure under local anaesthetic can help with micturition and menstruation. Reversing FGM during pregnancy is best done at 20 weeks. PMID- 16262172 TI - Bullies beware. PMID- 16262173 TI - Positive results. PMID- 16262174 TI - Access denied. PMID- 16262175 TI - Setting the pace. PMID- 16262176 TI - Wound care for patients with darkly pigmented skin. AB - This article examines the main skin or wound assessment parameters used for patients with darkly pigmented skin. Differences in skin pigmentation are discussed along with the challenges health professionals encounter in clinical practice and why linking theory to practice is vital. PMID- 16262177 TI - The role of genetic factors in maintaining health. AB - This article reviews the essential background information nurses need to help them understand how genetics influences health and ill-health. The typical human genetic make-up is described, along with an explanation of how changes to this can result in disease. The author also describes the characteristics of different patterns of inheritance. PMID- 16262179 TI - Heart failure. PMID- 16262178 TI - Treating severe psoriasis: an update. AB - This article provides an overview of the common skin condition psoriasis, and the various treatments that are available, which include the newly available biologic drugs for severe psoriasis. It examines quality of life issues, including the rejection and stigmatisation felt by many patients, and explores coping strategies that can help patients. PMID- 16262180 TI - New blood. PMID- 16262181 TI - Global decision. PMID- 16262182 TI - Money troubles. PMID- 16262183 TI - A test of the formal and modern theories of matching. AB - The present study tested a formal, or purely mathematical, theory of matching, and a modern account derived by McDowell (1986) that incorporates deviations from strict matching-bias and sensitivity. Six humans pressed a lever for monetary reinforcers on five concurrent variable interval (VI) schedules of reinforcement. All schedules were presented during each session. The magnitude on one alternative remained constant, and five magnitudes were presented across sessions on the other alternative. To test the formal account, two absolute response rate equations were fitted to the response and reinforcement rates at each alternative at each magnitude. Although the equations accounted for a high percentage of variance, there was a significant negative correlation between the standardized residuals and the predicted response rates. To test the modern account, an ensemble of four equations was fitted to the data. The equations predicted relative and absolute responding, and the independent variables in each equation were adjusted for bias and sensitivity. The equations accounted for a high percentage of variance, and the standardized residuals were not correlated with the predicted response rates. The values of the parameters were consistent with empirical findings and theoretical predictions, including the prediction that k should remain constant across changes in reinforcer magnitude. The results suggest that the formal theory of matching does not describe the data, and that the modern theory may provide an accurate and coherent description of concurrent and single-alternative responding. PMID- 16262184 TI - Associative symmetry in the pigeon after successive matching-to-sample training. AB - If an organism is explicitly taught an A-->B association, then might it also spontaneously learn the symmetrical B-->A association? Little evidence attests to such "associative symmetry" in nonhuman animals. We report for the first time a clear case of associative symmetry in the pigeon. Experiment 1 used a successive go/no go matching-to-sample procedure, which showed all of the training and testing stimuli in one location and intermixed arbitrary and identity matching trials. We found symmetrical responding that was as robust during testing (B-->A) as during training (A-->B). In Experiment 2, we trained different pigeons using only arbitrary matching trials before symmetry testing. No symmetrical responding was found. In Experiment 3, we trained other pigeons with only arbitrary matching trials and then tested for symmetry. When these pigeons, too, did not exhibit symmetrical responding, we retrained them with intermixed identity and arbitrary matching trials. Less robust symmetrical responding was obtained here than in Experiment 1. Collectively, these results suggest that identity matching may have to be learned concurrently with arbitrary matching from the outset of training for symmetry to emerge. PMID- 16262185 TI - Earning and obtaining reinforcers under concurrent interval scheduling. AB - Contingencies of reinforcement specify how reinforcers are earned and how they are obtained. Ratio contingencies specify the number of responses that earn a reinforcer, and the response satisfying the ratio requirement obtains the earned reinforcer. Simple interval schedules specify that a certain time earns a reinforcer, which is obtained by the first response after the interval. The earning of reinforcers has been overlooked, perhaps because simple schedules confound the rates of earning reinforcers with the rates of obtaining reinforcers. In concurrent variable-interval schedules, however, spending time at one alternative earns reinforcers not only at that alternative, but at the other alternative as well. Reinforcers earned for delivery at the other alternative are obtained after changing over. Thus the rates of earning reinforcers are not confounded with the rate of obtaining reinforcers, but the rates of earning reinforcers are the same at both alternatives, which masks their possibly differing effects on preference. Two experiments examined the separate effects of earning reinforcers and of obtaining reinforcers on preference by using concurrent interval schedules composed of two pairs of stay and switch schedules (MacDonall, 2000). In both experiments, the generalized matching law, which is based on rates of obtaining reinforcers, described responding only when rates of earning reinforcers were the same at each alternative. An equation that included both the ratio of the rates of obtaining reinforcers and the ratio of the rates of earning reinforcers described the results from all conditions from each experiment. PMID- 16262186 TI - Effects of adding a second reinforcement alternative: implications for Herrnstein's interpretation of r(e). AB - Herrnstein's hyperbola describes the relation between response rate and reinforcer rate on variable-interval (VI) schedules. According to Herrnstein's (1970) interpretation, the parameter r(e) represents the reinforcer rate extraneous to the alternative to which the equation is fitted (the target alternative). The hyperbola is based on an assumption that extraneous reinforcer rate remains constant with changes in reinforcer rate on the target alternative (the constant-r(e) assumption) and that matching with no bias and perfect sensitivity occurs between response and reinforcer ratios. In the present experiment, 12 rats pressed levers for food on a series of 10 VI schedules arranged on the target alternative. Across conditions, six VI values and extinction were arranged on a second alternative. Reinforcer rate on the second alternative, r2, negatively covaried with reinforcer rate on the target alternative for five of the six VI values on the second alternative, and significant degrees of bias and undermatching occurred in response ratios. Given covariation of reinforcer rate on the second and target alternatives, the constant-r(e) assumption can be maintained only by assuming that reinforcer rate from unmeasured background sources, rb, covaries with reinforcer rate on the second alternative such that their sum, r(e), remains constant. In a single schedule arrangement, however, r(e) equals rb and thus rb is assumed to remain constant, forcing a conceptual inconsistency between single- and concurrent schedule arrangements. Furthermore, although an alternative formulation of the hyperbola can account for variations in bias and sensitivity, the modified equation also is based on the constant-r(e) assumption and therefore suffers from the same logical problem as the hyperbola when reinforcer rate on the second alternative covaries with reinforcer rate on the target alternative. PMID- 16262188 TI - The effect of test schedules on the formation of linked perceptual classes. AB - After training conditional discriminations among selected stimuli from two perceptual classes, the emergence of novel relations involving other members of both classes was assessed using cross-class probes. The cross-class probes were presented using one of four different testing schedules. In the 2/9 test, nine different probes were presented in each of two test blocks. In the 6/3 test, three different probes were presented in each of six test blocks. In the 18/1-RND test, each of the 18 cross-class probes was presented in separate test blocks. In the 2/9 and 6/3 tests, the cross-class probes were presented in a randomized order within test block. In the 18/1-RND test, the cross-class probes were presented in a randomized sequence. In the 18/1-PRGM test, however, the cross class probes were presented in a programmed order (i.e., the values of the stimuli in each cross-class probe were changed systematically in the succession of probe presentations). About 55% of the linked perceptual classes emerged during the 2/9, 6/3, and 18/1-RND tests. Thus the number of different probes in a test block did not influence the emergence of classes as long as the probes were presented in a random order. Virtually all classes emerged during the 18/1-PRGM test. Thus at least one ordered introduction of different cross probes resulted in the reliable emergence of linked perceptual classes. Mechanisms responsible for linked perceptual class formation are discussed along with the relation of these classes to other complex categories. PMID- 16262187 TI - Crewmember performance before, during, and after spaceflight. AB - The development of technologies for monitoring the welfare of crewmembers is a critical requirement for extended spaceflight. Behavior analytic methodologies provide a framework for studying the performance of individuals and groups, and brief computerized tests have been used successfully to examine the impairing effects of sleep, drug, and nutrition manipulations on human behavior. The purpose of the present study was to evaluate the feasibility and sensitivity of repeated performance testing during spaceflight. Four National Aeronautics and Space Administration crewmembers were trained to complete computerized questionnaires and performance tasks at repeated regular intervals before and after a 10-day shuttle mission and at times that interfered minimally with other mission activities during spaceflight. Two types of performance, Digit-Symbol Substitution trial completion rates and response times during the most complex Number Recognition trials, were altered slightly during spaceflight. All other dimensions of the performance tasks remained essentially unchanged over the course of the study. Verbal ratings of Fatigue increased slightly during spaceflight and decreased during the postflight test sessions. Arousal ratings increased during spaceflight and decreased postflight. No other consistent changes in rating-scale measures were observed over the course of the study. Crewmembers completed all mission requirements in an efficient manner with no indication of clinically significant behavioral impairment during the 10-day spaceflight. These results support the feasibility and utility of computerized task performances and questionnaire rating scales for repeated measurement of behavior during spaceflight. PMID- 16262189 TI - Acquisition of cocaine self-administration with unsignaled delayed reinforcement in rhesus monkeys. AB - Six experimentally naive rhesus monkeys produced 0.01 mg/kg/infusion cocaine by lever pressing under a tandem fixed-ratio 1 differential-reinforcement-of-other behavior schedule. One lever press initiated an unsignaled 15- or 30-s delay culminating in cocaine delivery. Each press made during the delay reset the delay interval. With two exceptions, responding was acquired and maintained at higher rates than responding on a second (inoperative) lever. For the exceptions, a cancellation contingency was arranged in which each formerly inoperative-lever response reset the tandem schedule. This manipulation reduced presses on the inoperative lever. Subsequently, the consequences of responding on the two levers were reversed, and the monkeys again responded at higher rates on the operative lever. As a comparison, 3 additional experimentally naive monkeys received response-independent cocaine deliveries. Although lever pressing was observed, it extinguished and was subsequently reestablished under the tandem schedule. The results suggest that although response-reinforcer contiguity is not required for cocaine to acquire reinforcing functions, a response-reinforcer relation appears necessary. PMID- 16262191 TI - [Nursing care in ECMO]. PMID- 16262190 TI - A theory of attending and reinforcement in conditional discriminations. AB - A model of conditional discrimination performance (Davison & Nevin, 1999) is combined with the notion that unmeasured attending to the sample and comparison stimuli, in the steady state and during disruption, depends on reinforcement in the same way as predicted for overt free-operant responding by behavioral momentum theory (Nevin & Grace, 2000). The rate of observing behavior, a measurable accompaniment of attending, is well described by an equation for steady-state responding derived from momentum theory, and the resistance to change of observing conforms to predictions of momentum theory, supporting a key assumption of the model. When probabilities of attending are less than 1.0, the model accounts for some aspects of conditional-discrimination performance that posed problems for the Davison-Nevin model: (a) the effects of differential reinforcement on the allocation of responses to the comparison stimuli and on accuracy in several matching-to-sample and signal-detection tasks where the differences between the stimuli or responses were varied across conditions, (b) the effects of overall reinforcer rate on the asymptotic level and resistance to change of both response rate and accuracy of matching to sample in multiple schedules, and (c) the effects of fixed-ratio reinforcement on accuracy. Some tests and extensions of the model are suggested, and the role of unmeasured events in behavior theory is considered. PMID- 16262192 TI - [Do we need nursing chambers in Germany?]. PMID- 16262193 TI - [Problems in expert assessment of nursing care need in children]. PMID- 16262194 TI - [Preventive health is the emphasis of political career objectives of BeKD e.V]. PMID- 16262195 TI - [Violence in pediatric nursing]. PMID- 16262197 TI - [The optimiX brochure--revised recommendations for nutrition of children and adolescents]. PMID- 16262196 TI - [Organ donation in childhood--what should be done when and how?]. PMID- 16262198 TI - [Language development and promotion in the first 3 years of life]. PMID- 16262199 TI - [Developmentally determined readiness for drinking]. PMID- 16262200 TI - [Youth and tobacco: how to prevent habit adoption]. PMID- 16262201 TI - [Educators inform: voice disorders in children and adults. Revised leaflets on etiology, manifestations and treatment available]. PMID- 16262202 TI - [The mentally ill are human]. PMID- 16262203 TI - [How safe are ADHD drugs?]. PMID- 16262204 TI - [Applying a teaching plan outline in Rhineland-Westphalia promotes readiness for specialty education]. PMID- 16262205 TI - [Phenotypes]. PMID- 16262206 TI - Genetic bases for endophenotypes in psychiatric disorders. AB - This article reviews the concept of an endophenotype, with particular reference to heritability as well as diagnostic specificity. An endophenotype need not be heritable, for example, the possible influence of in utero viral infections for schizophrenia. However, heritability is a useful characteristic for a potential endophenotype, as it can be studied in relation to a plausible candidate gene. It should be noted that the traditional methods of demonstrating heritability, eg, twin studies, can be supplemented with DNA sequence studies, suggesting heritability. Endophenotypes need not be specific to a given nosological class of psychiatric disorders, as these classes do not reflect biological categories. Evidence for two useful schizophrenia endophenotypes, the P50 abnormalities and cognitive deficits, is summarized. PMID- 16262207 TI - Environmental programming of stress responses through DNA methylation: life at the interface between a dynamic environment and a fixed genome. AB - Early experience permanently alters behavior and physiology. These effects are, in part, mediated by sustained alterations in gene expression in selected brain regions. The critical question concerns the mechanism of these environmental "programming" effects. We examine this issue with an animal model that studies the consequences of variations in mother-infant interactions on the development of individual differences in behavioral and endocrine responses to stress in adulthood. Increased levels of pup licking/grooming by rat mothers in the first week of life alter DNA structure at a glucocorticoid receptor gene promoter in the hippocampus of the offspring. Differences in the DNA methylation pattern between the offspring of high- and low-licking/grooming mothers emerge over the first week of life; they are reversed with cross-fostering; they persist into adulthood; and they are associated with altered histone acetylation and transcription factor (nerve growth factor-induced clone A [NGFIA]) binding to the glucocorticoid receptor promoter. DNA methylation alters glucocorticoid receptor expression through modifications of chromatin structure. Pharmacological reversal of the effects on chromatin structure completely eliminates the effects of maternal care on glucocorticoid receptor expression and hypothalamic-pituitary adrenal (HPA) responses to stress, thus suggesting a causal relation between the maternally induced, epigenetic modification of the glucocorticoid receptor gene and the effects on stress responses in the offspring. These findings demonstrate that the structural modifications of the DNA can be established through environmental programming and that, in spite of the inherent stability of this epigenomic marker, it is dynamic and potentially reversible. PMID- 16262208 TI - The use of neurophysiological endophenotypes to understand the genetic basis of schizophrenia. AB - Specifying the complex genetic architecture of the "fuzzy" clinical phenotype of schizophrenia is an imposing problem. Utilizing metabolic, neurocognitive, and neurophysiological "intermediate" endophenotypic measures offers significant advantages from a statistical genetics standpoint. Endophenotypic measures are amenable to quantitative genetic analyses, conferring upon them a major methodological advantage compared with largely qualitative diagnoses using the Diagnostic and Statistical Manual of Mental Health, 4th Edition (DSM-IV). Endophenotypic deficits occur across the schizophrenia spectrum in schizophrenia patients, schizotypal patients, and clinically unaffected relatives of schizophrenia patients. Neurophysiological measures, such as P50 event-related suppression and the prepulse inhibition (PPI) of the startle response, are endophenotypes that can be conceptualized as being impaired because of a single genetic abnormality in the functional cascade of DNA to RNA to protein. The "endophenotype approach" is also being used to understand other medical disorders, such as colon cancer, hemochromatosis, and hypertension, where there is interplay between genetically conferred vulnerability and nongenetic stressors. The power and utility of utilizing endophenotypes to understand the genetics of schizophrenia is discussed in detail in this article. PMID- 16262209 TI - Endophenotypes in the personality disorders. AB - The identification of endophenotypes in the personality disorders may provide a basis for the identification of underlying genotypes that influence the traits and dimensions of the personality disorders, as well as susceptibility to major psychiatric illnesses. Clinical dimensions of personality disorders that lend themselves to the study of corresponding endophenotypes include affective instability, impulsivity, aggression, emotional information processing, cognitive disorganization, social deficits, and psychosis. For example, the propensity to aggression can be evaluated by psychometric measures, interview, laboratory paradigms, neurochemical imaging, and pharmacological studies. These suggest that aggression is a measurable trait that may be related to reduced serotonergic activity. Hyperresponsiveness of amygdala and other limbic structures may be related to affective instability, while structural and functional brain alterations underlie the cognitive disorganization in psychotic-like symptoms of schizotypal personality disorder. Thus, an endophenotypic approach not only provides clues to underlying candidate genes contributing to these behavioral dimensions, but may also point the way to a better understanding of pathophysiological mechanisms. PMID- 16262210 TI - Alcoholism: the dissection for endophenotypes. AB - Alcohol dependence (alcoholism) is a complex disorder attributed to the interaction of genetic and environmental factors that form a collage of "disease" predisposition, which is not identical for every alcohol-dependent individual. There is considerable evidence to demonstrate that genetic predisposition accounts for roughly half the risk in the development of alcohol dependence. Both family and population studies have identified a number of genomic regions with suggestive links to alcoholism, yet there have been relatively few definitive findings with regard to genetic determinants of alcoholism. This ambiguity can be attributed to a multitude of complications of studying complex mental disorders, such as clinical heterogeneity, polygenic determinants, reduced penetrance, and epistatic effects. Complex mental disorders are clinical manifestations described by combinations of various signs and symptoms. One approach to overcoming the ambiguity in studying the association between genetic risk factors and disease is to dissect the complex, heterogeneous disorder by using intermediate phenotypes- or endophenotypes--to generate more homogeneous diagnostic groupings than an all encompassing definition, such as the Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV)-derived term "alcohol dependence" or the commonly used term "alcoholism." The advantage of using endophenotypes is that the number of influential factors that contribute to these characteristics should be fewer and more easily identified than the number of factors affecting the heterogeneous entity of alcohol dependence (alcoholism). A variety of alcohol related characteristics have been investigated in epidemiological, clinical, and basic research as potential endophenotypes of alcohol dependence. These include phenotypes related to alcohol metabolism, physiological and endocrine measures, neural imaging, electrophysiology, personality, drinking behavior, and responses to alcohol and alcohol-derived cues. This review summarizes the current literature, focused on human data, of promising endophenotypes for dissecting alcoholism. PMID- 16262212 TI - How should clinicians interpret results reflecting the effect of an intervention on composite endpoints: should I dump this lump? PMID- 16262211 TI - Intermediate phenotypes in schizophrenia: a selective review. AB - Studies aiming to identify susceptibility genes for schizophrenia and other complex psychiatric disorders are faced with the confounds of subjective clinical criteria, commonly occurring phenocopies, significant between-subject variability of candidate traits, and the likelihood of allelic and locus heterogeneity that has been shown to define the genetics of other complex human brain and somatic disorders. Additionally, research aimed at identification of the molecular origins of schizophrenia must also deal with the confounding nature of the human brain. Unlike organs with a few common cellular phenotypes, transcriptomes, and proteomes, individual neurons are often distinct from one another in all of these respects. In this review, we present recent work testing the assumption that studies of genetic susceptibility in complex polygenic disorders such as schizophrenia might be enhanced by the identification of intermediate phenotypes related to more fundamental aspects of brain development and function. Progress in the identification of meaningful intermediate phenotypes in schizophrenia has been made possible by the advent of newer methods in cognitive neuroscience and neuroimaging, and the use of combined multimodal techniques. PMID- 16262213 TI - Prophylactic coronary artery revascularization before elective vascular surgery did not improve long-term survival. PMID- 16262214 TI - Radical prostatectomy reduced death and progression more than watchful waiting in early prostate cancer. PMID- 16262215 TI - Review: chemotherapy and hormonal therapy reduce recurrence and mortality at 15 years in early breast cancer. PMID- 16262216 TI - Review: tiotropium reduces exacerbations and hospitalizations in COPD and improves quality of life. PMID- 16262217 TI - Daily corticosteroids were not better than as-needed corticosteroids in mild persistent asthma. PMID- 16262218 TI - A varicella-zoster virus vaccine reduced the burden of illness of herpes zoster in older adults. PMID- 16262219 TI - Review: early tracheostomy is not better than late tracheostomy for reducing all cause mortality in critically ill patients. PMID- 16262220 TI - Amlodipine or lisinopril was not better than chlorthalidone for reducing CVD risk in hypertensive black or nonblack patients. PMID- 16262221 TI - Endovascular and open repair did not differ for mortality rates at 2 years in abdominal aortic aneurysm. PMID- 16262222 TI - Screening and active management reduced perinatal complications more than routine care in gestational diabetes. PMID- 16262223 TI - Review: alpha-glucosidase inhibitors improve glycemic control but have uncertain effects on patient-important outcomes in type 2 diabetes. PMID- 16262224 TI - A lifestyle intervention or metformin prevented or delayed the onset of the metabolic syndrome in persons at risk. PMID- 16262225 TI - Review: invasive management after unstable angina or non-ST-segment elevation MI does not reduce risk for death or MI. PMID- 16262226 TI - Review: routine invasive management after unstable angina or non-ST-segment elevation MI reduces risk for death or MI. PMID- 16262227 TI - Review: clinical evidence and consensus support the recommendation of 15 interventions in hip osteoarthritis. PMID- 16262228 TI - Review: both aerobic and home-based quadriceps strengthening exercises reduce pain and disability in knee osteoarthritis. PMID- 16262229 TI - Review: 700 to 800 IU/d of vitamin D reduces hip and nonvertebral fractures in older persons. PMID- 16262230 TI - Calcium and vitamin D supplementation did not reduce fractures in women >/= 70 years of age. PMID- 16262231 TI - Vitamin D3, calcium, or both did not prevent secondary fractures in elderly people. PMID- 16262232 TI - An immediate antiepileptic drug regimen reduced short-term (2-y) recurrence of seizures more than a deferred regimen. PMID- 16262233 TI - Review: prompt endoscopy is not a cost-effective strategy for initial management of dyspepsia. PMID- 16262234 TI - Review: paired quantitative blood cultures most accurately detect intravascular device-related bloodstream infection. PMID- 16262235 TI - Review: computed tomographic colonography has high specificity but low-to moderate sensitivity for detecting colorectal polyps. PMID- 16262236 TI - Administrative data feedback failed to improve quality of care in acute myocardial infarction. PMID- 16262237 TI - Simvastatin was cost-effective for reducing major vascular events in vascular disease or diabetes mellitus. PMID- 16262238 TI - An algorithm comprising 7 baseline variables predicted the 2-year work disability status in nonspecific back pain. PMID- 16262239 TI - Role of the unique peptide tail in hyperthermostable Aquifex aeolicus cochaperonin protein 10. AB - All known cochaperonin protein 10 (cpn10) molecules are heptamers of seven identical subunits noncovalently linked by beta-strand interactions. Cpn10 from the deep-branching, hyperthermophilic bacterium Aquifex aeolicus (Aacpn10) shows high homology with mesophilic and other thermophilic cpn10 sequences, except for a 25-residue C-terminal extension not found in any other cpn10. Prior to atomic structure information, we here address the role of the tail by biophysical means. A tail-lacking variant (Aacpn10-del25) also adopts a heptameric structure in solution and exhibits nativelike substrate-refolding activity. Thermal and chemical perturbations of both Aacpn10 and Aacpn10-del25, probed by far-UV circular dichroism, demonstrate that both proteins have high thermodynamic stability. Heptamer-monomer dissociation midpoints were defined by isothermal titration calorimetry; at 25 degrees C, the values for Aacpn10 and Aacpn10-del25 are within 2-fold of each other and close to reported midpoints for mesophilic cpn10 proteins. In contrast, the monomer stabilities for the A. aeolicus proteins are significantly higher than those of mesophilic homologues at 30 degrees C; thus, heptamer thermophily is a result of more stable monomers. Electron microscopy data reveals that Aacpn10-del25 heptamers are prone to stack on top of each other forming chainlike molecules; the electrostatic surface pattern of a structural model can explain this behavior. Taken together, the unique tail in Aacpn10 is not required for heptamer structure, stability, or function; instead, it appears to be an ancient strategy to avoid cochaperonin aggregation at extreme temperatures. PMID- 16262240 TI - Conformational heterogeneity at position U37 of an all-RNA hairpin ribozyme with implications for metal binding and the catalytic structure of the S-turn. AB - The hairpin ribozyme is an RNA enzyme that performs site-specific phosphodiester bond cleavage between nucleotides A-1 and G+1 within its cognate substrate. Previous functional studies revealed that the minimal hairpin ribozyme exhibited "gain-of-function" cleavage properties resulting from U39C or U39 to propyl linker (C3) modifications. Furthermore, each "mutant" displayed different magnesium-dependence in its activity. To investigate the molecular basis for these gain-of-function variants, crystal structures of minimal, junctionless hairpin ribozymes were solved in native (U39), and mutant U39C and U39(C3) forms. The results revealed an overall molecular architecture comprising two docked internal loop domains folded into a wishbone shape, whose tertiary interface forms a sequestered active site. All three minimal hairpin ribozymes bound Co(NH(3))(6)(3+) at G21/A40, the E-loop/S-turn boundary. The native structure also showed that U37 of the S-turn adopts both sequestered and exposed conformations that differ by a maximum displacement of 13 A. In the sequestered form, the U37 base packs against G36, and its 2'-hydroxyl group forms a water mediated hydrogen bond to O4' of G+1. These interactions were not observed in previous four-way-junction hairpin ribozyme structures due to crystal contacts with the U1A splicing protein. Interestingly, the U39C and U39(C3) mutations shifted the equilibrium conformation of U37 into the sequestered form through formation of new hydrogen bonds in the S-turn, proximal to the essential nucleotide A38. A comparison of all three new structures has implications for the catalytically relevant conformation of the S-turn and suggests a rationale for the distinctive metal dependence of each mutant. PMID- 16262241 TI - Variations in antigen-antibody association kinetics as a function of pH and salt concentration: a QSAR and molecular modeling study. AB - The relationship between three environmental factors (ionic strength, pH, and temperature) and antigen-antibody binding kinetics was investigated using QSAR (quantitative structure-activity relationship) and molecular modeling approaches. The interaction used for this analysis is that between the camel antibody fragment cAbLys3 and lysozyme. Binding kinetics were measured using a Biacore 2000 instrument, at NaCl concentrations between 50 and 500 mM, at pH's between 5 and 10, and at temperatures between 15 and 30 degrees C, according to multivariate experimental designs. Variations in kinetic on- and off-rate parameters were up to 400- and 16-fold, respectively. Mathematical models that relate log k(on) to experimental conditions were developed. They indicated an influence of all three factors, with a clear dependency between pH and NaCl concentration for their effect on k(on). These models were able to predict on rate parameters under new experimental conditions. Titration calculations using continuum electrostatics were performed on the crystallographic structures of the isolated and bound proteins to gain structural insight for the on-rate enhancement observed at pH <6.5 and low salt concentrations. These calculations rule out electrostatic steering linked to global and/or local charge variations in the molecules as the factor responsible for the on-rate enhancement at low pH. His 111 of cAbLys3, located at the binding interface, can adopt two side chain orientations with different intramolecular contacts. The results of the calculations suggest an alternative mechanism whereby the conformation of the interfacial His 111 depends on the charge, and these differences in conformation may influence the solvation energy and the subsequent binding kinetics. Our results stress the complex relationship between environmental conditions and molecular binding properties. PMID- 16262242 TI - Residue F4 plays a key role in modulating oxygen affinity and cooperativity in Scapharca dimeric hemoglobin. AB - Residue F4 (Phe 97) undergoes the most dramatic ligand-linked transition in Scapharca dimeric hemoglobin, with its packing in the heme pocket in the unliganded (T) state suggested to be a primary determinant of its low affinity. Mutation of Phe 97 to Leu (previously reported), Val, and Tyr increases oxygen affinity from 8- to 100-fold over that of the wild type. The crystal structures of F97L and F97V show side chain packing in the heme pocket for both R and T state structures. In contrast, in the highest-affinity mutation, F97Y, the tyrosine side chain remains in the interface (high-affinity conformation) even in the unliganded state. Comparison of these mutations reveals a correlation between side chain packing in the heme pocket and oxygen affinity, indicating that greater mass in the heme pocket lowers oxygen affinity due to impaired movement of the heme iron into the heme plane. The results indicate that a key hydrogen bond, previously hypothesized to have a central role in regulation of oxygen affinity, plays at most only a small role in dictating ligand affinity. Equivalent mutations in sperm whale myoglobin alter ligand affinity by only 5 fold. The dramatically different responses to mutations at the F4 position result from subtle, but functionally critical, stereochemical differences. In myoglobin, an eclipsed orientation of the proximal His relative to the A and C pyrrole nitrogen atoms provides a significant barrier for high-affinity ligand binding. In contrast, the staggered orientation of the proximal histidine found in liganded HbI renders its ligand affinity much more susceptible to packing contacts between F4 and the heme group. These results highlight very different strategies used by cooperative hemoglobins in molluscs and mammals to control ligand affinity by modulation of the stereochemistry on the proximal side of the heme. PMID- 16262243 TI - The C2A domain of synaptotagmin exhibits a high binding affinity for copper: implications in the formation of the multiprotein FGF release complex. AB - Human acidic fibroblast growth factor (hFGF-1) is a potent mitogen and is involved in the regulation of key cellular process such as angiogenesis, differentiation, and morphogenesis. hFGF-1 is a signal peptide-less protein that is released into the extracellular compartment as a multiprotein complex consisting of S100A13, synaptotagmin (Syt1), and a hFGF-1 homodimer. Cu(2+) is known to play an important role in the formation of the multiprotein release complex. The source of Cu(2+) required for the formation of the multiprotein release complex is not clear. In this study, we show that the cytoplasmic C2A domain of synaptotagmin binds to Cu(2+) ions with high affinity. Results from the isothermal calorimetry (ITC), near-UV circular dichroism (CD), and absorption spectroscopy experiments suggest that four Cu(2+) ions bind per molecule of C2A domain. Far-UV CD and limited trypsin digestion analysis reveal that the C2A domain undergoes a mild conformational change upon binding to Cu(2+). Competition experiments monitored by ITC and fluorescence resonance energy transfer indicate that Cu(2+) and Ca(2+) ions share common binding sites on the C2A domain. Cu(2+) ions compete with and replace Ca(2+) ions bound to the C2A domain. Two dimensional nuclear magnetic resonance spectroscopy data clearly show that Cu(2+) ions bind to the Ca(2+) binding sites in the loops (loops 1-3) located at the apex of the structure of the C2A domain. In addition, there is a unique Cu(2+) binding site located in the loop connecting beta-strands 7 and 8. It appears that the C2A domain provides the Cu(2+) ions required for the formation of the multiprotein FGF release complex. PMID- 16262244 TI - Phosphorylation of aquaporin PvTIP3;1 defined by mass spectrometry and molecular modeling. AB - The water channel protein PvTIP3;1 (alpha-TIP) is a member of the Major Intrinsic Protein membrane channel family. The in vitro activity of this aquaporin is dependent on phosphorylation, and the protein is phosphorylated in vivo by a membrane-associated Ca(2+)-dependent kinase. Mutagenesis studies have implicated three serine residues as kinase targets, but only phosphorylation of Ser7 has been observed in vivo. An atomic model of PvTIP3;1 generated by homology modeling suggested that Ser7 is the only residue that would be sterically accessible to kinases. To further explain the phosphorylation of PvTIP3;1, we overexpressed this aquaporin in the methylotrophic yeast Pichia pastoris and purified the hexahistidine-tagged protein by immobilized metal affinity chromatography. Mass spectrometry confirmed that a fraction of recombinant PvTIP3;1 was phosphorylated. Phosphatase and kinase treatments indicated that Ser7 was the only residue that could be phosphorylated. In addition, mass spectrometry indicated that the native and expressed proteins are N-terminally acetylated. This is the first demonstration that a full-length, recombinant aquaporin can be produced in yeast and authentically phosphorylated in vitro. Characterization of phosphorylation-mediated gating in PvTIP3;1 will serve as a paradigm for understanding gating mechanisms of other channels. PMID- 16262245 TI - Requirement of SRC-family tyrosine kinases in fat accumulation. AB - Src-family tyrosine kinases mediate many receptor signals to various biological responses. Here we investigate the requirement of Src-family tyrosine kinases in adipogenesis. The biochemical mechanism by which insulin induces adipogenesis, converting fibroblast cells to adipocytes, is not clear. We show that fibroblast cells deficient of three ubiquitously expressed Src-family members (Src, Yes, and Fyn), SYF cells, are refractory to hormonally induced fat accumulation. The defect is rescued by reintroduction of c-Src into SYF cells. Furthermore, Src family tyrosine kinases are required in the early steps of insulin signaling; it is responsible for the tyrosine phosphorylation of adaptor protein c-Cbl. Deficiency of c-Cbl blocked adipogenesis. These genetic and biochemical data clearly demonstrate that Src-family tyrosine kinases serve as a critical signal relay, via phosphorylation of c-Cbl, for fat accumulation, and provide potential new strategies for treating obesity. PMID- 16262246 TI - Specificity of binding of all-trans-retinyl ester to RPE65. AB - Membrane-bound RPE65 (mRPE65) is a binding protein for all-trans-retinyl esters, which are the substrates for the isomerization reaction that completes the visual cycle. RPE65 is essential for rhodopsin regeneration and, hence, for vision. As RPE65 appears to be part of the rate-limiting pathway in the visual cycle, specific antagonists of the molecule will be important in evaluating its full physiological role. The protein is known to stereoselectively bind all-trans retinyl esters (tREs), with dissociation constants in the 50 nM range. This study explores the overall binding specificity of RPE65 with respect to both retinoids and other isoprenoids in an effort to define the specificity of binding, and to begin the process of designing specific antagonists for it. The nature of the specificity directed toward the three main structural elements (retinoid, linker, and acyl moieties) in the tRE molecule is reported. In the all-trans-retinyl ester series, binding affinity increased as a function of the hydrophobicity of the fatty acyl group. In the linker region, binding affinities were little affected by amide, ketone, and ether replacements for the carboxy ester moiety of the naturally occurring tRE ligand. Finally, modifications in the all-trans retinoid moiety are also tolerated. For example, E,E-farnesyl palmitate binds with approximately the same affinity as does all-trans-retinyl palmitate. Other isoprenoid analogues also bind, as do truncated retinoids in the beta-ionone series. Therefore, mRPE65 is a moderately specific retinoid binding protein directed at long chain all-trans-retinyl esters. PMID- 16262247 TI - Binding of 5'-GMP to the GluR2 AMPA receptor: insight from targeted molecular dynamics simulations. AB - Guanine nucleotides behave as competitive antagonists at ionotropic glutamate receptors and show neuroprotective activity in different experimental excitotoxicity paradigms, both in vivo and in cultured cell preparations. Taking 5'-GMP as the reference nucleotide, we have tried to understand how these molecules interact with the agonist-binding site of the GluR2 alpha-amino-3 hydroxy-5-methyl-4-isoxazolepropionate (AMPA) receptor. Using a crystallographic model of the ligand-binding core of the GluR2 receptor in complex with kainate, we have previously analyzed the structural changes associated to the binding of agonists to the receptor and suggested a mechanism for the coupling of agonist binding to channel gating. In the present investigation we used the structure of the apo form of the receptor to probe the primary interactions between GMP and GluR2 by means of an automated docking program. A targeted molecular dynamics (TMD) simulation procedure was subsequently used to force the closing of the protein and to study the rearrangement of the ligand and surrounding amino acids. The resulting structure provides a plausible model of the nucleotide-receptor complex. Indirect support for the validity of our approach was obtained when the same methodology was shown to yield structures of the kainate-GluR2 and 6,7 dinitroquinoxaline-2,3-dione (DNQX)-GluR2 complexes that were in very good agreement with the published crystallographic structures. Both the stacking interaction between the phenyl ring of Tyr73 and the purine ring of GMP and a salt bridge between the phosphate group of GMP and Arg108 in the S1 domain, together with several hydrogen bonds, are proposed to secure the anchoring of GMP to the agonist-binding site. Unlike conventional competitive antagonists, such as DNQX, occupancy of the site by GMP still allows receptor segments S1 and S2 to close tightly around GMP without interacting with the critical residue Glu209 that triggers channel opening. Thus, GMP appears to be rather a false agonist than a competitive antagonist. This fact and the nature of the energy barriers that stabilize GMP bound to the closed form of the receptor provide an explanation for the unusual behavior of some guanine nucleotides in ligand displacement experiments. PMID- 16262248 TI - Characterization of the heparin/heparan sulfate binding site of the natural cytotoxicity receptor NKp46. AB - NKp46 is a member of a group of receptors collectively termed natural cytotoxicity receptors (NCRs) that are expressed by natural killer (NK) cells. NCRs are capable of mediating direct killing of tumor and virus-infected cells by NK cells. We have recently shown that NKp46 recognizes the heparan sulfate moieties of membranal heparan sulfate proteoglycans (HSPGs), thus enabling lysis of tumor cells by NK cells. In the current study, we further examined the residues in NKp46 that may be involved in heparan sulfate binding on tumor cells. On the basis of both the electrostatic potential map and comparison to the heparin binding site on human fibronectin, we predicted a continuous region containing the basic amino acids K133, R136, H139, R142, and K146 to be involved in NKp46 binding to heparan sulfate. Mutating these amino acids on NKp46D2 to noncharged amino acids retained its virus binding capacity but reduced its binding to tumor cells with a 10-100 fold lower K(D) when tested for direct binding to heparin. The minimal length of the heparin/heparan sulfate epitope recognized by NKp46 was eight saccharides as predicted from the structure and proven by testing heparin oligomers. Testing selectively monodesulfated heparin oligomers emphasized the specific contributions of O-sulfation, N-sulfation, and N-acetylation to epitope recognition by NKp46. The characterization of heparan sulfate binding region in NKp46 offers further insight into the identity of the ligands for NKp46 and the interaction of NK and cancers. PMID- 16262249 TI - Structural insights into enzyme regulation for inositol 1,4,5-trisphosphate 3 kinase B. AB - D-Myoinositol 1,4,5-trisphophate 3-kinases (IP(3)-3Ks) play important roles in metazoan cellular signaling. It has been demonstrated that mice without a functional version of IP(3)-3K isoform B are deficient in peripheral T-cells, indicating that IP(3)-3KB is essential to the developing immune system. The recent apo IP(3)-3KA structure exhibited a helix at the catalytic domain N terminus exhibited a helix at the N-terminus of the catalytic domain, with a tryptophan indole moiety mimicking the binding mode of the substrate ATP purine ring, suggesting a mechanism of autoinhibition. Here we present the structure of the complete catalytic domain of IP(3)-3KB, including the CaM binding domain in complex with Mg(2+) and ATP. The crystal structure reveals a homodimeric arrangement of IP(3)-3KB catalytic domains, mediated via an intermolecular antiparallel beta-sheet formed from part of the CaM binding region. Residues from the putative autoinhibitory helix are rearranged into a loop configuration, with extensive interactions with the bound ATP. Mutagenesis of residues from this region reveals that substitution of the putative autoinhibitory tryptophan generates a hyperactive enzyme which retains Ca(2+)/CaM sensitivity. The IP(3) 3KB structure suggests a mechanism of enzyme activation, and raises the possibility that an interaction between IP(3)-3KB molecules may occur as part of the catalytic or regulatory cycle. PMID- 16262250 TI - Molecular interactions of nonpeptide agonists and antagonists with the melanocortin-4 receptor. AB - The melanocortin-4 (MC4) receptor is a potential therapeutic target for obesity and cachexia, for which nonpeptide agonists and antagonists are being developed, respectively. The aim of this study was to identify molecular interactions between the MC4 receptor and nonpeptide ligands, and to compare the mechanism of binding between agonist and antagonist ligands. Nonpeptide ligand interaction was affected by mutations that reduce peptide ligand binding (D122A, D126A, S190A, M200A, F261A, and F284A), confirming overlapping binding determinants for peptide and nonpeptide ligands. The common halogenated phenyl group of nonpeptide ligands was a determinant of F261A and F284A mutations' affinity-reducing effect, implying this group interacts with the aromatic side chains of these residues. All affected compounds contain this group, the mutations reduced binding of 2,4 dichloro-substituted compounds more than 4-chloro-substituted-compounds, and F284A mutation eliminated the affinity-enhancing effect of 2-chloro-substitution. F261A and F284A mutations reduced the affinity of antagonists more than agonists, suggesting that the stronger ligand interaction with these residues, the lower the ligand efficacy. Supporting this hypothesis, F261A mutation increased the efficacy of nonpeptide antagonist and partial agonist ligands. D122A and D126A mutations reduced nonpeptide ligand interaction. Removing the ligands' derivatized amide group eliminated the effect of the mutations. Interaction of agonists, which bear a common amine within this group, was strongly reduced by D126A mutation (550-3300-fold), suggesting an electrostatic interaction between the amine and the acidic group of D126. These postulated interactions with aromatic and acidic regions of the MC4 receptor are consistent with a molecular model of the receptor. Furthermore, the strength of interaction with the aromatic pocket, and potentially the acidic pocket, controls the signaling efficacy of the ligand. PMID- 16262251 TI - Expression of functional G protein-coupled receptors in photoreceptors of transgenic Xenopus laevis. AB - G protein-coupled receptors (GPCRs) constitute the largest superfamily of transmembrane signaling proteins; however, the only known GPCR crystal structure is that of rhodopsin. This disparity reflects the difficulty in generating purified GPCR samples of sufficient quantity and quality. Rhodopsin, the light receptor of retinal rod neurons, is produced in large amounts of homogeneous quality in the vertebrate retina. We used transgenic Xenopus laevis to convert these retina rod cells into bioreactors to successfully produce 20 model GPCRs. The receptors accumulated in rod outer segments and were homogeneously glycosylated. Ligand and [(35)S]GTPgammaS binding assays of the 5HT(1A) and EDG(1) GPCRs confirmed that they were properly folded and functional. 5HT(1A)R was highly purified by taking advantage of the rhodopsin C-terminal immunoaffinity tag common to all GPCR constructs. We have also developed an automated system that can generate hundreds of transgenic tadpoles per day. This expression approach could be extended to other animal model systems and become a general method for the production of large numbers of GPCRs and other membrane proteins for pharmacological and structural studies. PMID- 16262252 TI - An isotope-edited FTIR investigation of the role of Ser-L223 in binding quinone (QB) and semiquinone (QB-) in the reaction center from Rhodobacter sphaeroides. AB - In the photosynthetic reaction center (RC) from the purple bacterium Rhodobacter sphaeroides, proton-coupled electron-transfer reactions occur at the secondary quinone (Q(B)) site. Several nearby residues are important for both binding and redox chemistry involved in the light-induced conversion from Q(B) to quinol Q(B)H(2). Ser-L223 is one of the functionally important residues located near Q(B). To obtain information on the interaction between Ser-L223 and Q(B) and Q(B)(-), isotope-edited Q(B)(-)/Q(B) FTIR difference spectra were measured in a mutant RC in which Ser-L223 is replaced with Ala and compared to the native RC. The isotope-edited IR fingerprint spectra for the C=O [see text] and C=C [see text] modes of Q(B) (Q(B)(-)) in the mutant are essentially the same as those of the native RC. These findings indicate that highly equivalent interactions of Q(B) and Q(B)(-) with the protein occur in both native and mutant RCs. The simplest explanation of these results is that Ser-L223 is not hydrogen bonded to Q(B) or Q(B)(-) but presumably forms a hydrogen bond to a nearby acid group, preferentially Asp-L213. The rotation of the Ser OH proton from Asp-L213 to Q(B)( ) is expected to be an important step in the proton transfer to the reduced quinone. In addition, the reduced quinone remains firmly bound, indicating that other distinct hydrogen bonds are more important for stabilizing Q(B)(-). Implications on the design features of the Q(B) binding site are discussed. PMID- 16262253 TI - Reaction mechanism and regulation of mammalian thioredoxin/glutathione reductase. AB - Thioredoxin/glutathione reductase (TGR) is a recently discovered member of the selenoprotein thioredoxin reductase family in mammals. In contrast to two other mammalian thioredoxin reductases, it contains an N-terminal glutaredoxin domain and exhibits a wide spectrum of enzyme activities. To elucidate the reaction mechanism and regulation of TGR, we prepared a recombinant mouse TGR in the selenoprotein form as well as various mutants and individual domains of this enzyme. Using these proteins, we showed that the glutaredoxin and thioredoxin reductase domains of TGR could independently catalyze reactions normally associated with each domain. The glutaredoxin domain is a monothiol glutaredoxin containing a CxxS motif at the active site, which could receive electrons from either the thioredoxin reductase domain of TGR or thioredoxin reductase 1. We also found that the C-terminal penultimate selenocysteine was required for transfer of reducing equivalents from the thiol/disulfide active site of TGR to the glutaredoxin domain. Thus, the physiologically relevant NADPH-dependent activities of TGR were dependent on this residue. In addition, we examined the effects of selenium levels in the diet and perturbations in selenocysteine tRNA function on TGR biosynthesis and found that expression of this protein was regulated by both selenium and tRNA status in liver, but was more resistant to this regulation in testes. PMID- 16262254 TI - Primary and secondary modes of DNA recognition by the NarL two-component response regulator. AB - NarL is a model response regulator for bacterial two-component signal transduction. The NarL C-terminal domain DNA binding domain alone (NarL(C)) contains all essential DNA binding determinants of the full-length NarL transcription factor. In the full-length NarL protein, the N-terminal regulatory domain must be phosphorylated to release the DNA binding determinants; however, the first NarL(C)-DNA cocrystal structure showed that dimerization of NarL(C) on DNA occurs in a manner independent of the regulatory domain [Maris, A. E., et al. (2002) Nat. Struct. Biol. 9, 771-778]. Dimerization via the NarL(C) C-terminal helix conferred high-affinity recognition of the tail-to-tail promoter site arrangement. Here, two new cocrystal structures are presented of NarL(C) complexed with additional 20mer oligonucleotides representative of other high affinity tail-to-tail NarL binding sites found in upstream promoter regions. DNA structural recognition properties are described, such as backbone flexibility and groove width, that facilitate NarL(C) dimerization and high-affinity recognition. Lys 188 on the recognition helix accommodates DNA sequence variation between the three different cocomplexes by providing flexible specificity, recognizing the DNA major groove floor directly and/or via bridging waters. The highly conserved Val 189, which enforced significant DNA base distortion in the first cocrystal structure, enforces similar distortions in the two new cocrystal structures. Recognition also is conserved for Lys 192, which hydrogen bonds to guanines at regions of high DNA helical writhe. DNA affinity measurements for model NarL binding sites, including those that did not cocrystallize, suggest a framework for explaining the diversity of heptamer site arrangement and orientation. PMID- 16262255 TI - Ubiquitination of p21Cip1/WAF1 by SCFSkp2: substrate requirement and ubiquitination site selection. AB - Multiple proteolytic pathways are involved in the degradation of the cyclin dependent kinase inhibitor p21(Cip1/WAF1). Timed destruction of p21(Cip1/WAF1) plays a critical role in cell-cycle progression and cellular response to DNA damage. The SCF(Skp2) complex (consisting of Rbx1, Cul1, Skp1, and Skp2) is one of the E3 ubiquitin ligases involved in ubiquitination of p21(Cip1/WAF1). Little is known about how SCF(Skp2) recruits its substrates and selects particular acceptor lysine residues for ubiquitination. In this study, we investigated the requirements for SCF(Skp2) recognition of p21(Cip1/WAF1) and lysine residues that are ubiquitinated in vitro and inside cells. We demonstrate that ubiquitination of p21(Cip1/WAF1) requires a functional interaction between p21(Cip1/WAF1) and the cyclin E-Cdk2 complex. Mutation of both the cyclin E recruitment motif (RXL) and the Cdk2-binding motif (FNF) at the N terminus of p21(Cip1/WAF1) abolishes its ubiquitination by SCF(Skp2), while mutation of either motif alone has minimal effects, suggesting either contact is sufficient for substrate recruitment. Thus, SCF(Skp2) appears to recognize a trimeric complex consisting of cyclin E-Cdk2 p21(Cip1/WAF1). Furthermore, we show that p21(Cip1/WAF1) can be ubiquitinated at four distinct lysine residues located in the carboxyl-terminal region but not two other lysine residues in the N-terminal region. Any one of these four lysine residues can be targeted for ubiquitination in the absence of the others in vitro, and three of these four lysine residues are also ubiquitinated in vivo, suggesting that there is limited specificity in the selection of ubiquitination sites. Interestingly, mutation of the carboxyl-terminal proline to lysine enables ubiquitin conjugation at the carboxyl terminus of the substrate both in vitro and in vivo. Thus, our results highlight a unique property of the ubiquitination enzymatic reaction in that substrate ubiquitination site selection can be remarkably diverse and occur in distinct spatial areas. PMID- 16262256 TI - Equilenin-derived DNA adducts to cytosine in DNA duplexes: structures and thermodynamics. AB - The drug Premarin is the most widely used formula for hormone replacement therapy. However, long-term exposure to estrogens from the Premarin drug increases the risk of breast cancer. Equilin and equilenin, major components of Premarin, are predominantly metabolized to 4-hydroxyequilenin (4-OHEN). The quinoids produced by 4-OHEN oxidation react with dG, dA, and dC to form unusual stable cyclic bulky adducts, with four stereoisomers identified for each base adduct. The 4-OHEN-dC adducts are most predominant. They are mutagenic in vitro and have been found in human tumor tissue. We have carried out molecular modeling and molecular dynamics simulations to investigate structures and thermodynamics of the four 4-OHEN-dC stereoisomeric adducts in DNA duplexes. Our results show that the structure of each stereoisomer adduct in duplex DNA is specifically governed by its unique stereochemistry. The bulky adducts, with an obstructed Watson-Crick edge and an equilenin ring system near perpendicular to the damaged cytosine, are located in the B-DNA major or minor groove, with the modified cytosine in the syn or anti conformation, respectively. The DNA duplex structures are distorted, in terms of Watson-Crick pairing at and near the lesion, stacking interactions, and groove dimensions. Stereochemistry determines the orientation of the equilenin rings with respect to the 5'- to 3'-direction of the modified strand, as well as the positioning of the equilenin moiety's methyl and hydroxyl groups for each stereoisomer. The unusual structures and the stereochemical effects underlie their biological processing as miscoding DNA lesions whose mutagenic properties may contribute to breast cancer. PMID- 16262257 TI - Hammerheads derived from sTRSV show enhanced cleavage and ligation rate constants. AB - The catalytic properties of the hammerhead ribozyme embedded in the (+) strand of the satellite tobacco ringspot viral genome are analyzed with the goal of obtaining the elemental rate constants of the cleavage (k(2)) and ligation (k( )(2)) steps. Two different chimeras combining the sTRSV (+) hammerhead and the well-characterized hammerhead 16 were used to measure the cleavage rate constant (k(2)), the rate of approach to equilibrium (k(obs) = k(2) + k(-)(2)), and the fraction of full-length hammerhead at equilibrium (k(-)(2)/k(2) + k(-)(2)). When compared to minimal hammerheads that lack the recently discovered loop I-loop II interaction, an extended format hammerhead derived from sTRSV studied here shows at least a 20-fold faster k(2) and a 1300-fold faster k(-)(2) at 10 mM MgCl(2). However, the magnesium dependence of the cleavage rate is not significantly changed. Thus, the enhanced cleavage of this hammerhead observed in vivo is due to its higher intrinsic rate and not due to its tighter binding of magnesium ions. The faster k(-)(2) of this hammerhead suggests that ligation may be used to form circular RNA genomes. This in vitro system will be valuable for experiments directed at understanding the hammerhead mechanism and the role of the loop I loop II interaction. PMID- 16262258 TI - Conformational dynamics of M2 helices in KirBac channels: helix flexibility in relation to gating via molecular dynamics simulations. AB - KirBac1.1 and 3.1 are bacterial homologues of mammalian inward rectifier K channels. We have performed extended molecular dynamics simulations (five simulations, each of >20 ns duration) of the transmembrane domain of KirBac in two membrane environments, a palmitoyl oleoyl phosphatidylcholine bilayer and an octane slab. Analysis of these simulations has focused on the conformational dynamics of the pore-lining M2 helices, which form the cytoplasmic hydrophobic gate of the channel. Principal components analysis reveals bending of M2, with a molecular hinge at the conserved glycine (Gly134 in KirBac1.1, Gly120 in KirBac3.1). More detailed analysis reveals a dimer-of-dimers type motion. The first two eigenvectors describing the motions of M2 correspond to helix kink and swivel motions. The conformational flexibility of M2 seen in these simulations correlates with differences in M2 conformation between that seen in the X-ray structures of closed channels (KcsA and KirBac) in which the helix is undistorted, and in open channels (e.g. MthK) in which the M2 helix is kinked. Thus, the simulations, albeit on a time scale substantially shorter than that required for channel gating, suggest a gating model in which the intrinsic flexibility of M2 about a molecular hinge is coupled to conformational transitions of an intracellular 'gatekeeper' domain, the latter changing conformation in response to ligand binding. PMID- 16262259 TI - Cooperative regulation of p70S6 kinase by receptor tyrosine kinases and G protein coupled receptors augments airway smooth muscle growth. AB - We have previously demonstrated that concomitant activation of receptor tyrosine kinases and certain G protein-coupled receptors (GPCRs) can promote a synergistic increase in the rate of airway smooth muscle cell (ASM) proliferation. Here we clarify the role of p70S6 kinase (p70S6K) as an integrator of receptor tyrosine kinase and GPCR signaling that augments ASM DNA synthesis by demonstrating that specific p70S6K phosphorylation sites receive distinct regulatory input from GPCRs that promotes sustained kinase activity critical to mitogenesis. Prolonged stimulation of ASM cells with EGF and thrombin induced a greater than additive effect in levels of p70S6K phosphorylated at residue T389, whereas a significant but more modest increase in the level of T229 and T421/S424 phosphorylation was also observed. The augmenting effects of thrombin could be dissociated from p42/p44 MAPK activation, as selective inhibition of thrombin-stimulated p42/p44 failed to alter the profile of cooperative p70S6K T389 phosphorylation, p70S6K kinase activity, or ASM [(3)H]thymidine incorporation. Thrombin stimulated a sustained increase in the level of Akt phosphorylation and also augmented EGF stimulated Akt phosphorylation. The cooperative effects of thrombin on Akt/p70S6K phosphorylation and [(3)H]thymidine incorporation were all attenuated by heterologous expression of Gbetagamma sequestrants. These data suggest that PI3K dependent T389/T229 phosphorylation is limiting in late-phase p70S6K activation by EGF and contributes to the cooperative effect of GPCRs on p70S6K activity and cell growth. PMID- 16262260 TI - Divergence in nonspecific hydrophobic packing interactions in the apo state, and its possible role in functional specialization of mitochondrial and microsomal cytochrome b5. AB - The outer mitochondrial membrane isoform of mammalian cytochrome b(5) (OM b(5)) is distinguished from the microsomal isoform (Mc b(5)) by its considerably greater stability. In contrast, OM and Mc apocytochrome b(5) (apo-b(5)) exhibit similar thermodynamic stability. Contributing substantially to the greater stability of OM b(5) relative to that of Mc b(5) is the presence of Leu at position 71. Replacing Leu-71 in OM b(5) with the corresponding Mc b(5) residue (Ser) not only diminishes holoprotein stability but also markedly compromises apoprotein stability. The studies reported herein were undertaken to clarify the role played by Leu-71 in stabilizing OM b(5)s relative to Mc b(5)s, and were motivated by the possibility that stability is related to other differences in OM and Mc b(5) properties that are important for their specialized subcellular roles. The results of these studies show that Leu-71 plays an essential role in maintaining the structural integrity of the heme-independent folding core of OM apo-b(5) (core 2), despite its location in the disordered empty heme-binding pocket (core 1). The conformational integrity of core 2 in Mc apo-b(5)s is not similarly dependent on the presence of a hydrophobic residue at position 71, providing new evidence for evolution of compensating structural features not present in OM b(5)s. We propose that Leu-71 achieves its effect on OM apo-b(5) core 2 structure by participating in a nonspecific hydrophobic collapse of disordered core 1, templated by more conformationally restricted side chains of residues in the beta-sheet that separates the two cores. We hypothesize that this has the added effect of maintaining core 1 of OM apo-b(5)s in a state more compact than that which occurs in Mc apo-b(5)s, possibly contributing to stronger heme binding by limiting the number of non-native conformations that the empty heme-binding pocket can populate. PMID- 16262261 TI - N-terminal stathmin-like peptides bind tubulin and impede microtubule assembly. AB - Microtubules are major cytoskeletal components involved in numerous cellular functions such as mitosis, cell motility, or intracellular traffic. These cylindrical polymers of alphabeta-tubulin assemble in a closely regulated dynamic manner. We have shown that the stathmin family proteins sequester tubulin in a nonpolymerizable ternary complex, through their stathmin-like domains (SLD) and thus contribute to the regulation of microtubule dynamics. We demonstrate here that short peptides derived from the N-terminal part of SLDs impede tubulin polymerization with various efficiencies and that phosphorylation of the most potent of these peptides reduces its efficiency as in full-length stathmin. To understand the mechanism of action of these peptides, we undertook a NMR-based structural analysis of the peptide-tubulin interaction with the most efficient peptide (I19L). Our results show that, while disordered when free in solution, I19L folds into a beta-hairpin upon binding to tubulin. We further identified, by means of saturation transfer difference NMR, hydrophobic residues located on the beta2-strand of I19L that are involved in its tubulin binding. These structural data were used together with tubulin atomic coordinates from the tubulin/RB3-SLD crystal structure to model the I19L/tubulin interaction. The model agrees with I19L acting through an autonomous tubulin capping capability to impede tubulin polymerization and provides information to help understand the variation of efficiency against tubulin polymerization among the peptides tested. Altogether these results enlighten the mechanism of tubulin sequestration by SLDs, while they pave the way for the development of protein-based compounds aimed at interfering with tubulin polymerization. PMID- 16262262 TI - Effect of lipid composition on the "membrane response" induced by a fusion peptide. AB - To understand the initial stages of membrane destabilization induced by viral proteins, the factors important for binding of fusion peptides to cell membranes must be identified. In this study, effects of lipid composition on the mode of peptides' binding to membranes are explored via molecular dynamics (MD) simulations of the peptide E5, a water-soluble analogue of influenza hemagglutinin fusion peptide, in two full-atom hydrated lipid bilayers composed of dimyristoyl- and dipalmitoylphosphatidylcholine (DMPC and DPPC, respectively). The results show that, although the peptide has a common folding motif in both systems, it possesses different modes of binding. The peptide inserts obliquely into the DMPC membrane mainly with its N-terminal alpha helix, while in DPPC, the helix lies on the lipid/water interface, almost parallel to the membrane surface. The peptide seriously affects structural and dynamical parameters of surrounding lipids. Thus, it induces local thinning of both bilayers and disordering of acyl chains of lipids in close proximity to the binding site. The "membrane response" significantly depends upon lipid composition: distortions of DMPC bilayer are more pronounced than those in DPPC. Implications of the observed effects to molecular events on initial stages of membrane destabilization induced by fusion peptides are discussed. PMID- 16262263 TI - Protein stabilization by introduction of cross-strand disulfides. AB - Disulfides cross-link residues in a protein that are separated in primary sequence and stabilize the protein through entropic destabilization of the unfolded state. While the removal of naturally occurring disulfides leads to protein destabilization, introduction of engineered disulfides does not always lead to significant stabilization of a protein. We have analyzed naturally occurring disulfides that span adjacent antiparallel strands of beta sheets (cross-strand disulfides). Cross-strand disulfides have recently been implicated as redox-based conformational switches in proteins such as gp120 and CD4. The propensity of these disulfides to act as conformational switches was postulated on the basis of the hypothesis that this class of disulfide is conformationally strained. In the present analysis, there was no evidence to suggest that cross strand disulfides are more strained compared to other disulfides as assessed by their torsional energy. It was also observed that these disulfides occur solely at non-hydrogen-bonded (NHB) registered pairs of adjacent antiparallel strands and not at hydrogen-bonded (HB) positions as suggested previously. One of the half-cystines involved in cross-strand disulfide formation often occurs at an edge strand. Experimental confirmation of the stabilizing effects of such disulfides was carried out in Escherichia coli thioredoxin. Four pairs of cross strand cysteines were introduced, two at HB and two at NHB pairs. Disulfides were formed in all four cases. However, as predicted from our analysis, disulfides at NHB positions resulted in an increase in melting temperature of 7-10 degrees C, while at HB positions there was a corresponding decrease of -7 degrees C. The reduced state of all proteins had similar stability. PMID- 16262264 TI - Investigation of the iron-sulfur cluster in Mycobacterium tuberculosis APS reductase: implications for substrate binding and catalysis. AB - The sulfur assimilation pathway is a key metabolic system in prokaryotes that is required for production of cysteine and cofactors such as coenzyme A. In the first step of the pathway, APS reductase catalyzes the reduction of adenosine 5' phosphosulfate (APS) to adenosine 5'-phosphate (AMP) and sulfite with reducing equivalents from the protein cofactor, thioredoxin. The primary sequence of APS reductase is distinguished by a conserved iron-sulfur cluster motif, -CC-X( approximately )(80)-CXXC-. Of the sequence motifs that are associated with 4Fe-4S centers, the cysteine dyad is atypical and has generated discussion with respect to coordination as well as the cluster's larger functional significance. Herein, we have used biochemical, spectroscopic, and mass spectrometry analysis to investigate the iron-sulfur cluster and its role in the mechanism of Mycobacterium tuberculosis APS reductase. Site-directed mutagenesis of any cysteine residue within the conserved motif led to a loss of cluster with a concomitant loss in catalytic activity, while secondary structure was preserved. Studies of 4Fe-4S cluster stability and cysteine reactivity in the presence and absence of substrates, and in the free enzyme versus the covalent enzyme intermediate (E-Cys-S-SO(3)(-)), suggest a structural rearrangement that occurs during the catalytic cycle. Taken together, these results demonstrate that the active site functionally communicates with the iron-sulfur cluster and also suggest a functional significance for the cysteine dyad in promoting site differentiation within the 4Fe-4S cluster. PMID- 16262266 TI - DNA-induced dimerization of poly(ADP-ribose) polymerase-1 triggers its activation. AB - In response to DNA strand breaks in the genome of higher eukaryotes, poly(ADP ribose)polymerase 1 (PARP-1) catalyses the covalent attachment of ADP-ribose units from NAD(+) to various nuclear acceptor proteins including PARP-1 itself. This post-translational modification affecting proteins involved in chromatin architecture and in DNA repair plays a critical role in cell survival as well as in caspase-independent cell death. Although PARP-1 has been best-studied for its role in genome stability, several recent reports have demonstrated its role in the regulation of transcription. In this study, fluorescence spectroscopy and biochemical techniques are used to investigate the association of the amino terminal DNA-binding domain of human PARP-1 (hPARP-1 DBD) with various DNA substrates, characterized by different DNA ends and sequence features (5'- or 3' recessed end, double strands, telomeric repeats, and the palindromic sequence of a Not I restriction site). The correlation between the binding mode of hPARP-1 DBD to the DNA oligoduplexes and the enzymatic activation of hPARP-1 is analyzed. We show that hPARP-1 DBD binds a 5'-recessed DNA end cooperatively with a stoichiometry of two proteins per DNA molecule. In contrast, a 1:1 stoichiometry is found in the presence of a 3'-recessed end and double-strand DNA. A palindromic structure like the Not I restriction site is shown to induce protein dimerization and high enzymatic activation, suggesting that it can represent a recognition element for hPARP-1 in undamaged cells. Protein dimerization is found to be a requisite for high enzymatic activity. Taken together, our data allow further characterization of the features of hPARP-1 recognition in damaged cells and bring additional evidence that hPARP-1 may also play a role in undamaged cells. PMID- 16262265 TI - Tropolysin, a new oligopeptidase from African trypanosomes. AB - Oligopeptidases are emerging as important pathogenic factors and therapeutic targets in trypanosome infections. We describe here the purification, cloning, and biochemical analysis of a new oligopeptidase from two pathogenic African trypanosomes. This oligopeptidase, which we have called tropolysin (encoded by the trn gene), represents an evolutionarily distant member of the M3A subfamily of metallopeptidases, ancestral to thimet oligopeptidase, neurolysin, and saccharolysin. The trn gene was present as a single copy per haploid genome, was expressed in both the mammalian and insect stages of the parasite life cycle, and encoded an 84 kDa protein. Both purified and hyperexpressed tropolysin hydrolyzed bradykinin-derived fluorogenic peptide substrates at restricted sites, with an alkaline pH optimum, and were activated by dithiothreitol and reduced glutathione and by divalent metal cations, in the order Zn(2+) > Co(2+) > Mn(2+). Under oxidizing conditions, tropolysin reversibly formed inactive multimers. Tropolysin exhibited a preference for acidic amino acid side chains in P(4), hydrophobic side chains in P(3), and hydrophobic or large uncharged side chains in P(1), P(1)', and P(3)', while the S(2)' site was unselective. Highly charged residues were not tolerated in P(1)'. Tropolysin was responsible for the bulk of the kinin degrading activity in trypanosome lysates, potently (k(cat) approximately 119 s( )(1)) inactivated the vasoactive kinins bradykinin and kallidin, and generated angiotensin(1-7) from angiotensin I. This hydrolysis both abolished the capacity of bradykinin to stimulate the bradykinin B(2) receptor and abrogated bradykinin prohypotensive properties in vivo, raising the possibility that tropolysin may play a role in the dysregulated kinin metabolism observed in the plasma of trypanosome-infected hosts. PMID- 16262267 TI - Gelatin binding to the 6F1(1)F2(2)F2 fragment of fibronectin is independent of module-module interactions. AB - Fibronectin, a large modular protein, interacts with many other proteins in the extracellular matrix and on the cell surface. It has previously been shown that interactions between noncontiguous modules exist in the collagen binding region. It is shown here that the interaction between the sixth type I module ((6)F1) and the second type II module ((2)F2) can be disrupted by mutation of a residue in the intermodule interface of the (6)F1(1)F2(2)F2 fragment. The perturbation of the interface and the binding of collagen-derived peptides to individual modules were assessed by high-resolution nuclear magnetic resonance (NMR) spectroscopy. Cooperativity between the modules in binding ligand was assessed by analytical gelatin affinity chromatography of the mutant and wild-type proteins. Differential scanning calorimetry (DSC) was used to probe the influence of the interface on module stability. It is shown that while the (6)F1-(2)F2 interface confers significant thermal stability to the (2)F2 module, it has little effect on gelatin binding activity of the (6)F1(1)F2(2)F2 fragment. PMID- 16262269 TI - Cocklebur-shaped colloidal dispersions. AB - Unique cocklebur-shaped colloidal dispersions were prepared using a combination of a nanoextruder applied to the aqueous solution containing methyl methacrylate (MMA) and n-butyl acrylate (n-BA) with azo-bis-isobutyronitrile (AIBN) or potassium persulfate (KPS) initiators and stabilized by a mixture of sodium dioctyl sulfosuccinate (SDOSS) and 1,2-bis(10,12-tricosadiynoyl)-sn-glycero-3 phosphocholine (DCPC) phospholipid. Upon extrusion and heating to 75 degrees C, methyl methacrylate/n-butyl acrylate (MMA/nBA) colloidal particles containing tubules pointing outward were obtained as a result of DCPC phospholipids present at the particle surfaces. The same cocklebur-shaped particles were obtained when classical polymerization was used without a nanoextruder under similar compositional and thermal conditions, giving a particle size of 159 nm. However, when Ca(2+) ions are present during polymerization, cocklebur morphologies are disrupted. Because DCPC tubules undergo a transition at 38 degrees C, such cocklebur morphologies may offer numerous opportunities for devices with stimuli responsive characteristics. PMID- 16262268 TI - Total chemical synthesis of human psoriasin by native chemical ligation. AB - Human psoriasin (S100A7), a member of the S100 family of calcium-binding proteins, is richly expressed in keratinocytes of patients suffering from psoriasis. To date, the exact physiological function of psoriasin abundant in many human cell types remains unclear. A recent report by Schroder and colleagues suggests that psoriasin, purified from human stratum corneum extracts, selectively kills Escherichia coli by sequestering Zn(2+) ions essential for bacterial growth, indicative of an important role in innate immune defense against microbial infection. We chemically synthesized the N-terminally acetylated psoriasin of 100 amino acid residues using solid phase peptide synthesis in combination with native chemical ligation. More than 140 mg of highly pure and correctly folded synthetic psoriasin was obtained from a single synthesis on a 0.25 mmol scale. Analysis of synthetic psoriasin by size exclusion chromatography showed that the protein forms a homodimer in solution. Circular dichroism analysis indicated that the alpha-helicity of psoriasin increases by more than 20% in the presence of CaCl(2) or ZnCl(2), suggesting a metal ion binding induced conformational change. Circular dichroism based titration further established that the synthetic protein binds two Ca(2+) and two Zn(2+) ions per dimer, in agreement with the published structural findings. Importantly, the ability of the synthetic protein to kill E. coli and the inhibition of the killing by ZnCl(2) is comparable to that of psoriasin isolated from its natural source. The robust synthetic access to large quantities of human psoriasin should facilitate studies of its biological functions as well as its mode of action. PMID- 16262270 TI - Hydration of CO adsorbed on Pt(100) at cryogenic temperatures in UHV environments. AB - Adsorption of small amounts of D(2)O (ca. 0.01 L exposure) on CO(sat)/Pt(100) surfaces in ultrahigh vacuum at 105 K was found to split the asymmetric peak at 2100 cm(-1) in the infrared reflection absorption spectra attributed to the stretching of CO adsorbed on atop sites into two clearly defined features: an intense component, which shifted toward lower energies due to surface hydration of adsorbed CO (originally at 2100 cm(-1), peak a), and a smaller peak centered at 2094 cm(-1) (peak b), which remained fixed in position even after closing the D(2)O dosing valve. The energies of peaks a and b, as determined by statistical analyses, correlated very well with those reported in the literature for CO adsorbed at high coverages on Pt(100) originally in the (5 x 20) or (hex) reconstruction, and on the unreconstructed Pt(100)-(1 x 1), respectively, at 90 K. On these bases, the asymmetry of the peak observed for CO(sat)/Pt(100) (no D(2)O dosing) is ascribed to the presence of CO linearly adsorbed on these two different sites on the surface, for which the rate of hydration is larger for the (5 x 20) compared to the (1 x 1) phases. PMID- 16262271 TI - Ambient-pressure vapor deposition of octanethiol self-assembled monolayers. AB - Scanning tunneling microscopy was used to characterize self-assembled monolayers (SAMs) of octanethiol on Au(111), created using vapor deposition at elevated temperatures and ambient pressure. Monolayers contained large, close-packed ( radical3 x radical3)R30 degrees domains with sizes considerably larger than those typically formed from conventional solution-phase preparation and with crystallographically straight domain boundaries. New striped surface phases were also observed, including a 13 x radical3 phase with a density that was 69% of the close-packed density; these striped phases appeared topographically higher in STM images than close-packed monolayers. PMID- 16262272 TI - Ring stain effect at room temperature in silver nanoparticles yields high electrical conductivity. AB - We demonstrate that metallic rings formed spontaneously at room temperature via evaporation of aqueous drops containing silver nanoparticles (20-30 nm in diameter) exhibit high electrical conductivity (up to 15% of that for bulk silver). The mechanism underlying this self-assembly phenomena is the "ring stain effect", where self-pinning is combined with capillary flow to form a ring consisting of close-packed metallic nanoparticles along the perimeter of a drying droplet. Our macroscopic and microscopic (applying conductive atomic force microscopy) transport measurements show that the conductivity of the ring, which has a metallic brightness, is orders of magnitude larger than that of corresponding aggregates developed without the ring formation, where high conductivity is known to appear only after annealing at high temperature. PMID- 16262273 TI - Gelation of colloidal crystals without degradation in their transmission quality and chemical tuning. AB - A single-domain colloidal crystal with high transmission quality, prepared by a shear-induced process, was fixed as a hydrogel film by photopolymerization. Upon gelation, the original optical quality was almost perfectly preserved. By replacing the solvent, the gelled crystal could be converted to smaller lattice constant crystals without significant degradation in its transmission characteristics. The conversion results in a stop-band wavelength coverage across the entire visible light range. PMID- 16262274 TI - Role of the spacer of cationic gemini amphiphiles in the condensation of DNA. AB - The condensation of calf thymus DNA into the cholesteric-like psi-phase was observed by circular dichroism in liposome suspensions formulated with specific cationic gemini surfactants. The stereochemistry of the gemini spacer, the presence of specific functional groups, and the covalent link between the headgroups are fundamental issues in the condensation of DNA. Transmission electron microscopy images showed a multilamellar morphology, which corresponds with condensation. PMID- 16262275 TI - Synthesis of monodisperse biodegradable microgels in microfluidic devices. AB - Microgels are promising materials in drug delivery and biomedicine. Although monodisperse microgels would offer considerable advantages, most microgels investigated and used today are polydisperse in size. We report on the fabrication of 10 mum sized monodisperse microgels by emulsifying an aqueous dextran-hydroxyethyl methacrylate (dex-HEMA) phase within an oil phase at the junction of microfluidic channels. Dex-HEMA microgels are biodegradable and are ideally suited for the controlled delivery of proteins. PMID- 16262276 TI - Reversible transformations of gold nanoparticle morphology. AB - Herein is reported a metamorphosis taking place in a gold nanosized system. The observed phenomenon of shape and size transformations was found to be completely reversible. Unlike most procedures in the literature where shape and size control occur in the synthetic step by adding growth- and shape-controlling agents such as surfactants or polymers, in this system postsynthetic changes in shape and size can be carried out simply by changing the ratio of reactive, competing reagents, more specifically, alkylthiols versus tetralkylammonium salts. Interestingly, the transfer of gold metal occurs (large prismatic particles to small particles and vice versa) under the influence of reagents that do not cause such interactions with bulk gold. All intermediate steps of the morphology change were observed using HRTEM and electron diffraction. The processes of breaking down and "welding back" solid metal nanoparticles occur under mild conditions and are remarkable examples of the unique chemical properties of nanomaterials. The described process is expected to be relevant to other nanoscale systems where similar structural circumstances could occur. PMID- 16262277 TI - Colloidal particles coated and stabilized by DNA-wrapped carbon nanotubes. AB - Single-walled carbon nanotubes (SWNTs) are dispersed in water via wrapping with short segments of single-stranded DNA (ssDNA). Small angle neutron scattering suggests a power-law exponent that is consistent with clustered nanotubes and hence marginal stability. The SWNT-ssDNA complex is used to stabilize dispersions of hydrophilic colloidal particles with the nanotubes adhered to the surface of the colloids. Near-infrared fluorescence microscopy demonstrates the interfacial band-gap fluorescence of these SWNT-coated particles, suggesting potential routes to novel platforms and applications. PMID- 16262278 TI - Fabrication of self-organized chemically and topologically heterogeneous patterns on the surface of polystyrene-b-oligothiophene block copolymer films. AB - A novel fabrication of the chemically and topologically heterogeneous patterns on the surface of polymeric films over an area of more than 1 square centimeter in a single step was demonstrated by using the self-organizing character of polystyrene-b-oligothiophene block copolymers. Hexagonally arranged open pores of a size of approximately 2 mum are spontaneously formed by casting the polymer solution under a moist air flow. The amphiphilic character of the polystyrene-b oligothiophene block copolymers played the crucial role as a surfactant to stabilize the inverse emulsion of water droplets in the organic solvent, and subsequently the structure of the arranged hydrophilic oligothiophene segments remained on the interiors of the micropores. The chemical composition of the surface of the microporous films was characterized by time-of-flight secondary ion mass spectrometry (ToF-SIMS) to prove the chemical heterogeneity. The ToF SIMS imaging clearly indicated that the oligothiophene forms the aggregated structure on the interior of the open micropores on the surface while the flat area on the surface was covered with the polystyrene. PMID- 16262280 TI - Hierarchical nanoparticle/block copolymer surface features via synergistic self assembly at the air-water interface. AB - This work demonstrates the first example of the controlled organization of semiconducting nanoparticles (NPs) using amphiphilic block copolymer self assembly at the air-water interface. Preferential interactions between polystyrene-functionalized NPs and the polystyrene block of an amphiphilic polystyrene-b-poly(ethylene oxide) block copolymer result in synergistic self assembly at the air-water interface, forming a range of highly stable one dimensional NP/polymer surface features, including branched nanowires, nanocables up to 100 microm in length, and nanowires with nanoring connectors. This strategy offers new routes to hierarchical hybrid assemblies with potential photonics applications because the nanoscale organization of NPs is coupled to features with dimensions that are commensurate with optical wavelengths. PMID- 16262279 TI - Characterization of micropatterned lipid membranes on a gold surface by surface plasmon resonance imaging and electrochemical signaling of a pore-forming protein. AB - We report the fabrication and characterization of a micropatterned membrane electrode for electrochemical signaling of a bacterial pore-forming toxin, Streptolysin O (SLO) from S. pyogenes. Microcontact printing of an alkylthiol monolayer was used to fabricate an array template, onto which cholesterol containing DMPC vesicles were fused to form lipid layer structures. The construction of the supported membranes, including pattern transfer and vesicle fusion, was characterized by in-situ surface plasmon resonance (SPR) imaging and electrochemistry. Quantitative analysis of the resulting membrane by using SPR angular shift measurements indicates that the membranes in the hydrophilic pockets have an average thickness of 8.2 +/- 0.4 nm. Together with fluorescence microscopy studies, the results suggest that this could be a mixed lipid assembly that may consist of a bilayer, vesicle fragments, and lipid junctions. The voltammetric response of the redox probe ferrocene carboxylic acid (FCA) was measured to quantify the toxin action on the supported membrane. The electrochemical measurements indicate that fusion of vesicles on the template blocked the access of FCA, whereas the injection of SLO toxin restored the redox response. The anodic peak current of FCA was found to increase with toxin concentration until a plateau was reached at 40 HU/mL. The method is highly sensitive such that 0.1 HU/mL of SLO (1.25 pM) can yield a well-defined response. In addition, it eliminates the need for a highly insulating layer in membrane sensing, which opens up new avenues in developing novel sensing interfaces for membrane-targeting proteins and peptides. PMID- 16262281 TI - Self-assembled, mesoporous polymeric networks for patterned protein arrays. AB - A facile, self-assembly approach to the fabrication of a robust, mesoporous, biocompatible polymeric network for the spatial organization of proteins is described. Surface-deposited poly(styrene) (PS) beads that assemble into a two dimensional (2-D) hexagonal array are used to template cross-linked poly(vinyl alcohol) (PVA), yielding an inverse opal structure. The porous, water insoluble network is used to entrain a model, soluble protein, green fluorescent protein (GFP). The polymeric network is characterized by atomic force microscopy (AFM) and optical microscopy, and the spatial localization of the incorporated GFP is determined by fluorescence microscopy. The results demonstrate that this system may constitute a versatile platform for the lateral organization of biomolecules. PMID- 16262282 TI - Drug transport in responding lipid membranes can be regulated by an external osmotic gradient. AB - In this paper, we demonstrate, for the first time, how an external osmotic gradient can be used to regulate diffusion of solutes across a lipid membrane. We present experimental and theoretical studies of the transport of different solutes across a monoolein membrane in the presence of an external osmotic gradient. The osmotic gradient introduces phase transformations in the membrane, and it causes nonlinear transport behavior. The external gradient can thus act as a kind of switch for diffusive transport in the skin and in controlled release drug formulations. PMID- 16262283 TI - In-plane enyne metathesis and subsequent Diels-Alder reactions on self-assembled monolayers. AB - We report in-plane enyne metathesis and subsequent Diels-Alder reactions on self assembled monolayers (SAMs) terminating in vinyl and acetylenyl groups on gold. After the formation of SAMs of vinyl and acetylenyl group-containing dithiols on gold, in-plane enyne metathesis of the vinyl and acetylenyl groups, leading to the formation of 1,3-diene, was achieved on the SAMs, and Diels-Alder reactions were then successfully performed with tetracyanoethylene, maleic anhydride, and maleimide. The reactions were confirmed by FT-IR spectroscopy, X-ray photoelectron spectroscopy, and time-of-flight secondary-ion mass spectrometry. In-plane enyne metathesis developed herein would offer a versatile platform for the functionalization of surfaces with mild reaction conditions and a high compatibility in functional groups. PMID- 16262284 TI - Template-induced enhanced ordering under confinement. AB - We report a surprisingly strong ordering of Si-(CH(3))(2) groups upon confinement between two surfaces, an oxidized poly(dimethyl siloxane) (PDMS(ox)) elastomer and a methyl-terminated self-assembled monolayer (octadecyltrichlorosilane (OTS)) on sapphire substrates. This enhanced ordering is induced by the template of ordered methyl groups of OTS and is not observed for other surfaces (fluorinated monolayers and sapphire substrates). This strong ordering is reminiscent of layering observed for confined symmetric molecules between two mica surfaces but was expected to vanish between rough macroscopic surfaces. These results provide new insights on confined structure at the interface between two solids and are important in the understanding of surface-controlled processes of practical importance. PMID- 16262285 TI - Preparation temperature dependence of size and polydispersity of alkylthiol monolayer protected gold clusters. AB - The influence of preparation temperature on the size and size distribution of dodecylthiol monolayer protected gold clusters was studied. The monolayer protected clusters (MPCs) were synthesized by two different variations of the Brust-Schiffrin procedure. In all of the experiments, the stoichiometry of the reactants dodecylthiol, HAuCl(4), and sodium borohydride was kept constant, while the temperature was varied in the range of -18 to +90 degrees C. Two series were performed in which an aqueous solution of NaBH(4) was either added over 30 s or all in one portion. The size and size distribution of the MPCs were determined by small-angle X-ray scattering (SAXS) and transmission electron microscopy (TEM). It has been demonstrated that in general the MPC size increases with elevated preparation temperatures. PMID- 16262286 TI - Flat orientation of hydrophobic cores induced by two-dimensional confinement of flexible bolaamphiphiles at the air-water interface. AB - We designed and synthesized a new bolaamphiphile consisting of a biphenyl core, flexible trisiloxane spacers, and terminal ammonium groups. The compression of the trisiloxane-containing bolaamphiphile at the air-water interface led to the formation of monolayer films with the hydrophobic rigid core lying flat on the film surface. Such monolayer structures were formed through the compression induced conformational change of the flexible bolaamphiphile from an extended state to a folded one as confirmed by surface pressure-area isotherm measurements, water contact angle measurements, atomic force microscopy, and UV vis spectroscopy. PMID- 16262287 TI - Mechanically responsive films of variable hydrophobicity made of polyelectrolyte multilayers. AB - Mechanically responsive surfaces that allow to switch reversibly from a hydrophobic to a hydrophilic substrate are reported. The surfaces are constituted of polyelectrolyte multilayers deposited on modified charged silicone sheets. n bilayers of poly(allylamine)-Nafion (PAH-Naf) and m bilayers of poly(allylamine) poly(acrylic acid) (PAH-PAA) composed the multilayers. A (PAH-Naf)(n) film possesses a water contact angle of around 105 degrees, whereas the contact angle of a (PAH-Naf)(4)-(PAH-PAA)(m) multilayer is around 50 degrees. When such a film with m < 5 and terminated by PAA is stretched out, its water contact angle increases up to around 100 degrees. Successive elongation/retraction cycles allow the water contact angle to alternate reversibly between 100 and 57 degrees indicating the reversible mechanical responsive nature of the film. PMID- 16262288 TI - Preparation of nanoporous MgO-coated TiO2 nanoparticles and their application to the electrode of dye-sensitized solar cells. AB - Sol-gel-derived Mg(OH)(2) gel was coated onto TiO(2) nanoparticles, and the subsequent thermal topotactic decomposition of the gel formed a highly nanoporous MgO crystalline coating. The specific surface area of the electrode that was prepared from the core-shell-structured TiO(2) nanoparticles significantly increased compared with that of the uncoated TiO(2) electrode. The increase in the specific surface area of the MgO-coated TiO(2) electrode was attributed to the highly nanoporous MgO coating layer that resulted from the topotactic reaction. Dye adsorption behavior and solar cell performance were significantly enhanced by employing the MgO-coated TiO(2) electrode. Optimized coating of a MgO layer on TiO(2) nanoparticles enhanced the energy conversion efficiency as much as 45% compared to that of the uncoated TiO(2) electrode. This indicates that controlling the extrinsic parameters such as the specific surface area is very important to improve the energy conversion efficiency of TiO(2)-based solar cells. PMID- 16262289 TI - A tribute to the phospholipid. PMID- 16262290 TI - Donnan effects in the steady-state diffusion of metal ions through charged thin films. AB - The steady-state diffusion of metals ions through thin films with fixed charged groups was investigated using diffusive gradients in thin films (DGT) measurements. Copolymers of acrylamide and sodium acrylate cross-linked with N,N' methylenebisacrylamide were used as diffusive gels. The rate of diffusion of cadmium ions through the gels was measured by determining the mass of cadmium bound to a backing chelex resin after a known deployment time. Variation of the ionic strength as well as the fixed charge density and the thickness of the gel layer allowed evaluation of the impact of the Donnan partitioning and the diffusion layer in solution on the observed steady-state flux of ions through the layer. The results underscore that, as the Donnan partitioning increases, the impact of the diffusion layer in solution becomes more significant. At modest Donnan potentials, Donnan partitioning controls the net flux of metal ions, whereas at conditions of increasing Donnan potential, i.e., at decreasing ionic strength, the flux is increasingly limited by diffusion in solution. An analytical expression is developed to describe the influence of Donnan partitioning on the observed steady-state flux of metal ions. PMID- 16262291 TI - Lipid bilayer disruption by polycationic polymers: the roles of size and chemical functional group. AB - Polycationic polymers are used extensively in biology to disrupt cell membranes and thus enhance the transport of materials into the cell. The highly polydisperse nature of many of these materials makes obtaining a mechanistic understanding of the disruption processes difficult. To design an effective mechanistic study, a monodisperse class of polycationic polymers, poly(amidoamine) (PAMAM) dendrimers, has been studied in the context of supported dimyristoylphosphatidylcholine (DMPC) lipid bilayers using atomic force microscopy (AFM). Aqueous solutions of amine-terminated generation 7 (G7) PAMAM dendrimers caused the formation of 15-40-nm-diameter holes in lipid bilayers. This effect was significantly reduced for smaller G5 dendrimers. For G3, no hole formation was observed. In addition to dendrimer size, surface chemistry had a strong influence on dendrimer-lipid bilayer interactions. In particular, acetamide-terminated G5 did not cause hole formation in bilayers. In all instances, the edges of bilayer defects proved to be points of highest dendrimer activity. A proposed mechanism for the removal of lipids by dendrimers involves the formation of dendrimer-filled lipid vesicles. By considering the thermodynamics, interaction free energy, and geometry of these self-assembled vesicles, a model that explains the influence of polymer particle size and surface chemistry on the interactions with lipid membranes was developed. These results are of general significance for understanding the physical and chemical properties of polycationic polymer interactions with membranes that lead to the transport of materials across cell membranes. PMID- 16262292 TI - Spectroscopic investigations of metalloporphyrin-oligopeptide systems: evidence for peptide aggregation. AB - Anionic pentapeptides consisting of a string of four glutamic acid residues terminated by either tyrosine (Glu4Tyr) or tryptophan (Glu4Trp) were synthesized, and their aggregation properties in buffered (pH = 7.0) aqueous solutions were investigated using two different approaches. In the first approach, the effects of the concentration of peptide used as its own probe (intrinsic probe) on its fluorescence emission, circular dichroism, surface tension, and solution pH yielded similar critical peptide concentrations of around 175 microM. This particular concentration was taken as evidence for peptide aggregation. In the second approach, peptide aggregation was investigated using cationic metalloporphyrins, tetrakis(N-methyl-4-pyridyl)porphyrin (Pd(II)TMPyP(4+) and Zn(II)TMPyP(4+)), as extrinsic probes. The effect of peptide concentration on porphyrin ground-state absorption confirmed peptide aggregation, but at a lower critical peptide concentration near 125 microM. This difference was attributed to the possible distortion introduced by the association of one (or more) large metalloporphyrin molecule with the peptide aggregates. Evidence for peptide aggregation was also demonstrated from the effect of peptide concentration on Pd(II)TMPyP(4+) triplet-state decay. The fast component (k(f), associated with electron transfer from the target Tyr and Trp residues to the porphyrin triplet state) was found to be independent of peptide concentration, implying no noticeable effect of peptide aggregation on the electron-transfer event. This was attributed to the fact that species formed by excitation of porphyrin associated with ion-pair complexes or bound to peptide aggregates and the diffusion together of the separate T(1) and peptide entities in the bulk phase are kinetically similar. On the other hand, the slower component (k(s)) of the decay, which is associated with the diffuse formation of an encounter complex between the free peptide and T(1) porphyrin (bulk phase), was peptide-dependent and displayed a critical peptide concentration near 125 microM, above which it became practically independent of peptide concentration. This invariance of k(s) was taken as an indication that the free peptide concentration in the bulk phase remains constant above 125 microM, the concentration at which peptide molecules prefer to associate as aggregates. PMID- 16262295 TI - Changes in the shape and mobility of colloidal gold nanorods with electrospray and differential mobility analyzer methods. AB - The potential of the electrospray technique in analyzing the structure of nonspherical colloidal particles that are below 100 nm in volume-equivalent diameter was demonstrated by online size measurement using a differential mobility analyzer (DMA) with a condensation nucleus counter (CNC) system. The measured mobility of gold nanorods was confirmed by electron microscope images and the theoretical calculation of particle mobility using the dynamic shape factor and slip correction factor. To evaluate the mobility, rod particles were modeled as both a cylinder and a prolate spheroid. This study also showed that the organic surfactant coated on rod particles might be removed and that the rod particles became spherical upon the elevation of the ambient temperature during the gas-phase dispersion of colloidal nanoparticles. Moreover, the thickness of the surfactants coated on rod particles was estimated by comparing the theoretically and experimentally obtained mobilities. PMID- 16262294 TI - Modeling of ultralyophobicity: suspension of liquid drops by a single asperity. AB - With the aim of understanding the underlying physical phenomenon associated with utlralyophobic (or super repellent) surfaces, model studies have been performed on single asperities of different size and shape. A small liquid drop was deposited on top of each model asperity, and liquid was sequentially added. If the advancing contact angle was sufficiently large, it was possible to suspend large drops atop asperities with an apparent contact angle approaching 180 degrees. If more and more liquid was added, eventually the suspended drops collapsed. Roughening the surface of the asperities further bolstered suspension. Using an analysis that accounts for both capillary forces and the influence of gravity, the critical suspension volume was correctly predicted for each liquid/asperity combination. PMID- 16262293 TI - Spontaneously formed vesicles of sodium N-(11-acrylamidoundecanoyl)-glycinate and L-alaninate in water. AB - Two N-acyl amino acid surfactants, sodium N-(11-acrylamidoundecanoyl)-glycinate (SAUG) and L-alaninate (SAUA), were synthesized and characterized in aqueous solution. A number of techniques, such as surface tension, fluorescence probe, light scattering, and transmission electron microscopy were employed for characterization of the amphiphiles in water. The surface and interfacial properties were measured. The amphiphiles have two critical aggregation concentrations. The results of surface tension and fluorescence probe studies suggested formation of bilayer self-assemblies in dilute aqueous solutions of the amphiphiles. The magnitudes of free energy change of aggregation have indicated that bilayer formation is more favorable in the case of SAUG. Steady-state fluorescence measurements of pyrene and 1,6-diphenyl-1,3,5-hexatriene (DPH) were used to study the microenvironment of the molecular self-assemblies. Temperature dependent fluorescence anisotropy change of DPH probe revealed phase transition temperature of the bilayer self-assemblies. The effects of pH on the structure of the self-assemblies of SAUG and SAUA have been studied. The role of intermolecular hydrogen bonding between amide groups upon aggregation toward microstructure formation in solution has been discussed. Circular dichroism spectra suggested the presence of chiral aggregates in an aqueous solution of SAUA. The transmission electron micrographs revealed the presence of closed spherical vesicles in aqueous solutions of the amphiphiles. Dynamic light scattering measurements were performed to obtain average size of the aggregates. PMID- 16262296 TI - Specialist gelator for ionic liquids. AB - Cyclo(l-beta-3,7-dimethyloctylasparaginyl-L-phenylalanyl) (1) and cyclo(L-beta-2 ethylhexylasparaginyl-L-phenylalanyl) (2), prepared from L-asparaginyl-L phenylalanine methyl ester, have been found to be specialist gelators for ionic liquids. They can gel a wide variety of ionic liquids, including imizazolium, pyridinium, pyrazolidinium, piperidinium, morpholinium, and ammonium salts. The mean minimum gel concentrations (MGCs) necessary to make gels at 25 degrees C were determined for ionic liquids. The gel strength increased at a rate nearly proportional to the concentration of added gelator. The strength of the transparent gel of 1-butylpyridinium tetrafluoroborate ([C(4)py]BF(4)), prepared at a concentration of 60 g L(-1) (gelator 1/[C(4)py]BF(4)), was ca. 1500 g cm( 2). FT-IR spectroscopy indicated that a driving force for gelation was intermolecular hydrogen bonding between amides and that the phase transition from gel to liquid upon heating was brought about by the collapse of hydrogen bonding. The gels formed from ionic liquids were very thermally stable; no melting occurs up to 140 degrees C when the gels were prepared at a concentration of 70 g L(-1) (gelator/ionic liquid). The ionic conductivities of the gels were nearly the same as those of pure ionic liquids. The gelator had electrochemical stability and a wide electrochemical window. When the gels were prepared from ionic liquids containing propylene carbonate, the ionic conductivities of the resulting gels increased to levels rather higher than those of pure ionic liquids. The gelators also gelled ionic liquids containing supporting electrolytes. PMID- 16262297 TI - Comparison of microenvironments of aqueous sodium dodecyl sulfate micelles in the presence of inorganic and organic salts: a time-resolved fluorescence anisotropy approach. AB - Microenvironments of aqueous sodium dodecyl sulfate (SDS) micelles was examined in the presence of additives such as sodium chloride and p-toluidine hydrochloride (PTHC) by monitoring the fluorescence anisotropy decays of two hydrophobic probes, 2,5-dimethyl-1,4-dioxo-3,6-diphenylpyrrolo[3,4-c]pyrrole (DMDPP) and coumarin 6 (C6). It has been well-established that SDS micelles undergo a sphere-to-rod transition and that their mean hydrodynamic radius increases from 19 to 100 A upon the addition of 0.0-0.7 M NaCl at 298 K. A similar size and shape transition is induced by PTHC at concentrations that are 20 times lower compared to that of NaCl. This study was undertaken to find out how the microviscosity of the micelles is influenced under these circumstances. It was noticed that the microviscosity of the SDS/NaCl system increased by approximately 45%, whereas there was a less than 10% variation in the microviscosity of the SDS/PTHC system. The large increase in the microviscosity of the former system with salt concentration has been rationalized on the basis of the high concentration of sodium ions in the headgroup region of the micelles and their ability to strongly coordinate with the water present in this region, which decreases the mobility of the probe molecules. PMID- 16262298 TI - Amino Acid based cationic surfactants in aqueous solution: physicochemical study and application of supramolecular chirality in ketone reduction. AB - The present study provides a molecular understanding of the origin of the chirality in aqueous micelles and its correlation with the proficiency of stereoselective ketone reduction. The effects of varied headgroup architecture on the surface-active properties as well as on other microstructural parameters were studied and correlated to the structural differences of these naturally occurring amino acid containing surfactants (1-4). Micropolarity sensed by pyrene showed that the micelles prepared using 1-4 are mostly hydrated; particularly large headgroup size surfactant produces more polar environment. A theoretical study was done to quantify the varied spatial dissymmetry for all four surfactants. Asymmetric reduction of prochiral ketones was carried out at the aqueous micellar interface of these chiral amphiphiles by exploiting the supramolecular chirality as evidenced from a circular dichroism study. The enantioselectivity of the reduction process is rationally improved through increase in spatial dissymmetry and steric constraint imposed at the micellar interface by the polar head of surfactants. PMID- 16262299 TI - Synthesis and characterization of N,N-dimethyldodecylamine-capped Aucore-Pdshell nanoparticles in toluene. AB - In this paper a convenient route for synthesizing Au(core)-Pd(shell) bimetallic nanoparticles in toluene has been reported as a result of co-reduction of gold(III) and palladium(II) precursors in toluene. N,N-Dimethyldodecylamine was used as a capping agent for the core-shell particles, which not only imparts stability to the organosol but also controls morphology of the evolved particles. The particles were characterized using UV-visible, transmission electron microscopy, and X-ray diffraction measurements. All results substantiate the formation of core-shell structure of the synthesized particles. PMID- 16262300 TI - Sugar-based gemini surfactants with peptide bonds-synthesis, adsorption, micellization, and biodegradability. AB - The sugar-based gemini surfactant with peptide bonds, N,N'-bisalkyl-N,N'-bis[2 (lactobionylamide)ethyl]hexanediamide (2C(n)peLac, in which n represents hydrocarbon chain lengths of 12 and 16), was synthesized by reacting adipoyl chloride with the corresponding monomeric surfactant N-alkyl-N' lactobionylethylenediamine (C(n)peLac), which was obtained by reacting ethylenediamine with alkyl bromide and lactobionic acid. The adsorption and micellization properties of C(n)peLac and 2C(n)peLac were characterized by the measurement of their equilibrium and dynamic surface tension, steady-state fluorescence using pyrene as a probe, dynamic light scattering (DLS), and time resolved fluorescence quenching (TRFQ), and their biodegradability was also investigated. The critical micelle concentration (cmc) decreases with an increase in the hydrocarbon chains from monomeric to gemini surfactants, whereas it increases with an increase in the chain length from 12 to 16 for both systems. The increases in both the hydrocarbon chain and the chain length of sugar-based surfactants reduce surface activities such as the ability to lower the surface tension, the occupied area per molecule, and the adsorption rate at the air/water interface. The sugar-based surfactants C(n)peLac and 2C(n)peLac exhibit unique aggregation behavior in aqueous solution. The DLS results indicate that the apparent hydrodynamic diameter of C(n)peLac micelles decreases sharply with increasing concentration, whereas that of 2C(n)peLac micelles decreases gradually. From the TRFQ measurement, it was observed that, as concentration increases, the aggregation numbers are almost constant for C(n)peLac, whereas they increase for 2C(n)peLac. These results imply that loosely packed micelles formed by sugar-based surfactants become tightly packed micelles as the concentration increases. Furthermore, it was found that 2C(n)peLac shows lower biodegradability than does C(n)peLac because it contains tertiary amines in the molecule. PMID- 16262301 TI - Fabrication of spherical colloidal crystals using electrospray. AB - We demonstrated the use of electrohydrodynamic atomization to prepare uniform sized emulsion droplets in which equal spheres of silica or polystyrene were dispersed. The size of the emulsion droplets was easily controlled by the electric field strength and the flow rate, independently of the diameter of the nozzles. During the evaporation of solvent in the droplets, spherical colloidal crystals were formed by self-assembly of the monodisperse colloidal spheres. The diameter of the spherical colloidal crystals was in the range of 10-40 microm. Depending on the stability of colloidal particles, the morphology of the self assembled structure was varied. In particular, silica spheres in ethanol droplets were self-assembled into compactly packed silica colloidal crystals in spherical shapes, whereas polystyrene latex spheres in toluene droplets self-assembled into spherical colloidal crystal shells with hollow cores. The silica colloidal assemblies reflected diffraction colors according to the three-dimensionally ordered arrangement of silica spheres. PMID- 16262302 TI - Sedimentation and phase transitions of colloidal gibbsite platelets. AB - We study the competition between sedimentation, gelation, and liquid crystal formation in suspensions of colloidal gibbsite platelets of five different sizes at three ionic strengths. For large particles (with diameters of 350, 420, and 570 nm) sedimentation is initially the most important factor determining the macroscopic behavior. Only after the main part of the sample has sedimented in an amorphous phase, phase separation takes place. For the smallest particles (diameter 210 and 270 nm), it is the other way around: fast (within one week) phase separation or gelation takes place, after which sedimentation determines the final macroscopic appearance. We distinguish six different scenarios within this two-fold scheme and interpret these on the basis of the previously obtained phase diagram of colloidal gibbsite platelets (van der Beek, D.; Lekkerkerker, H. N. W. Langmuir 2004, 20, 8582). PMID- 16262304 TI - Phase behavior of concentrated aqueous solutions of cetyltrimethylammonium bromide (CTAB) and sodium hydroxy naphthoate (SHN). AB - We have characterized the phase behavior of mixtures of the cationic surfactant cetyltrimethylammonium bromide (CTAB) and the organic salt 3-sodium-2-hydroxy naphthoate (SHN) over a wide range of surfactant concentrations using polarizing optical microscopy and X-ray diffraction. A variety of liquid crystalline phases, such as hexagonal, lamellar with and without curvature defects, and nematic, are observed in these mixtures. At high temperatures the curvature defects in the lamellar phase are annealed gradually on decreasing the water content. However, at lower temperatures these two lamellar structures are separated by an intermediate phase, where the bilayer defects appear to order into a lattice. The ternary phase diagram shows a high degree of symmetry about the line corresponding to equimolar CTAB/SHN composition, as in the case of mixtures of cationic and anionic surfactants. PMID- 16262303 TI - Peptoads, a group of amphiphilic long-chain triamides. AB - Eleven triamides bearing long alkyl chains have been synthesized to produce a new class of amphiphilic compounds (dubbed "peptoad"). The properties of these molecules have been investigated by X-ray analysis, solubility studies, light and electron microscopy, surface tensiometry, light scattering, drug dissolution, and molecular dynamics. In the solid state, the peptoads assemble in layers with both intra- and interlayer hydrogen bonding coupled to side-by-side proximity of the hydrocarbon chains. Peptoads with a terminal primary amide and a total of three amide NH sites are water-insoluble owing presumably to attractive forces in the solid state. However, peptoads with terminal -CONMe(2) groups and two internal amide NH sites are water-soluble at room temperature. This solubility is critically dependent upon the chain length. For example, a C(7)-chained peptoad is 1600 times more soluble than its C(9) analogue. High concentrations (6-8 M) of C(7) peptoads in water are clear and do not gel. Light microscopy shows long fibers floating in an isotropic liquid. Water-soluble peptoads are highly surface active, lowering water's surface tension as effectively as a soap with a much longer chain. Surface tension plots show a "critical aggregation concentration", but it is believed from light scattering and molecular dynamics that the aggregates grow continuously as more peptoad is added to the water. In answer to the inevitable (but valid) question, "What possible good are they?", it can be pointed out that a peptoad solubilizes a water-insoluble drug, paclitaxel (Taxol), as efficiently as does Cremophor EL, a commercial excipient widely used with paclitaxel and other nonpolar drugs. Peptoads, being small molecules and consisting of hydrolyzable amide groups, are likely biodegradable and less prone to the hypersensitivity and neurotoxicity found with Cremophor EL. PMID- 16262305 TI - The dramatic effect of architecture on the self-assembly of block copolymers at interfaces. AB - Dramatic morphological changes are observed in the Langmuir-Blodgett (LB) film assemblies of poly(ethylene glycol)-b-(styrene-r-benzocyclobutene) block copolymer (PEG-b-(S-r-BCB)) after intramolecular cross-linking of the S-r-BCB block to form a linear-nanoparticle structure. To isolate architectural effects and allow direct comparison, the linear block copolymer precursor and the linear nanoparticle block copolymer resulting from selective intramolecular cross linking of the BCB units were designed to have exactly the same molecular weight and chemical composition but different architecture. It was found that the effect of architecture is pronounced with these macromolecular isomers, which self assemble into dramatically different surface aggregates. The linear block copolymer forms disklike surface assemblies over the range of compression states, while the linear-nanoparticle block copolymer exhibits long (>10 microm) wormlike aggregates whose length increases as a function of increasing cross-linking density. It is shown that the driving force behind the morphological change is a combination of the altered molecular geometry and the restricted degree of stretching of the nanoparticle block because of the intramolecular cross-linking. A modified approach to interpret the pi-A isotherm, which includes presence of the block copolymer aggregates, is also presented, while the surface rheological properties of the block copolymers at the air-water interface provide in-situ evidence of the aggregates' presence at the air-water interface. PMID- 16262306 TI - Nanosecond imaging of microboiling behavior on pulsed-heated au films modified with hydrophilic and hydrophobic self-assembled monolayers. AB - Fast transient microboiling has been characterized on modified gold microheaters using a novel laser strobe microscopy technique. Microheater surfaces of different hydrophobicity were prepared using self-assembled monolayers of hexadecane thiol (hydrophobic) and 16-mercaptohexadecanol (hydrophilic) as well as the naturally hydrophilic bare gold surface. The microheater was immersed in a pool of water, and a 5-micros voltage pulse to the heater was applied, causing superheating of the water and nucleation of a vapor bubble on the heater surface. Light from a pulsed Nd:Yag laser was configured to illuminate and image the sample through a microscope assembly. The timing of the short duration (7.5 ns) laser flash was varied with respect to the voltage pulse applied to the heater to create a series of images illuminated by the flash of the laser. These images were correlated with the transient resistance change of the heater both during and after the voltage pulse. It was found that hydrophobic surfaces produced a bubble that nucleated at an earlier time, grew more slowly to a smaller maximum size, and collapsed more rapidly than bubbles formed on hydrophilic surfaces. PMID- 16262307 TI - Transition from homogeneous Langmuir-Blodgett monolayers to striped bilayers driven by a wetting instability in octadecylsiloxane monolayers. AB - We show that two dips of an oxidized silicon substrate through a prepolymerized n octadecylsiloxane monolayer at an air-water interface in a rapid succession produces periodic, linear striped patterns in film morphology extending over macroscopic area of the substrate surface. Langmuir monolayers of n octadecyltrimethoxysilane were prepared at the surface of an acidic subphase (pH 2) maintained at room temperature (22 +/- 2 degrees C) under relative humidities of 50-70%. The substrate was first withdrawn at a high dipping rate from the quiescent aqueous subphase (upstroke) maintained at several surface pressures corresponding to a condensed monolayer state and lowered soon after at the same rate into the monolayer covered subphase (downstroke). The film structure and morphology were characterized using a combination of optical microscopy, imaging ellipsometry, and Fourier transform infrared spectroscopy. An extended striped pattern, perpendicular to the pushing direction of the second stroke, resulted for all surface pressures when the dipping rate exceeded a threshold value of 40 mm min(-1). Below this threshold value, uniform deposition characterizing formation of a bimolecular film was obtained. Under conditions that favored striped deposition during the downstroke through the monolayer-covered interface, we observed a periodic auto-oscillatory behavior of the meniscus. The stripes appear to be formed by a highly correlated reorganization and/or exchange of the first monolayer, mediated by the Langmuir monolayer at the air-water interface. This mechanism appears distinctly different from nanometer scale stripes observed recently in single transfers of phospholipid monolayers maintained near a phase boundary. The stripes further exhibit wettability patterns useful for spatially selective functionalization, as demonstrated by directed adsorptions of an organic dye (fluorescein) and an oil (hexadecane). PMID- 16262308 TI - Formation of a freely suspended membrane via a combination of interfacial reaction and wetting. AB - Applying poly(ethoxysiloxane) (a liquid non-water-soluble polymer that can be hydrolyzed and cross-linked by diluted acids) to an air/pH 1 water interface gave rise to thin homogeneous solid layers. These layers were strong enough to be transferable to electron microscopy grids with holes of dimensions up to 150 microm and covered the holes as freely suspended membranes. No homogeneous layers were formed at an air/pH 5 water interface. Brewster angle microscopy images show that the poly(ethoxysiloxane) is not spontaneously forming a wetting layer on water. It initially forms lenses, which slowly spread out within several hours. We conclude that the spreading occurs simultaneously with the hydrolysis and cross-linking of the poly(ethoxysiloxane) and that the reaction products finally assist the complete wetting of the water surface. PMID- 16262309 TI - Adsorption structures on metals--a thermodynamic description. AB - Thermodynamic conditions of the adsorption of gaseous molecules on metals are discussed in this work. The equilibria of the formation of surface arrays comprising several atoms, the formation of ordered structures, and the dissolution of gas atoms in the metal have been reviewed. It was found that the heat and the free energy of chemisorption is affected by the change in the number and energy of bonds between metal atoms and the adsorbate in the structures being formed. It was pointed out that the change in the interactions between the surface metal atoms before and after the adsorption must be followed by an energetic effect that would affect the chemisorption energy. PMID- 16262310 TI - Diffusion of hydrosilanes from the control layer to the vinylsilane-rich flow membrane during the fabrication of microfluidic chips. AB - During the fabrication of poly(dimethylsiloxane) (PDMS)-based microfluidic chips, polymethylhydrosiloxane (PMHS) species in the control layer diffuse into the flow membrane, which contains polymethylvinylsiloxane (PMVS), and the components cross link together to form the mechanically enhanced membrane. The diffusion course was investigated by using attenuated total reflectance FTIR and the improvement of mechanical properties of the flow membrane was studied by measuring the Young's modulus and the tensile strength. PMID- 16262311 TI - Changes in hydration of lanthanide ions on binding to DNA in aqueous solution. AB - The interaction of the trivalent lanthanides Ce(III), Eu(III), and Tb(III) with sodium deoxyribonucleic acid (DNA) in aqueous solution has been studied using their luminescence spectra and decays. Complexation with DNA is indicated by changes in luminescence intensity. In the system terbium(III)-DNA, changes in luminescence with pH are suggested to be due to the protonation of phosphate groups. The degree of hydration of Tb(III) on binding to DNA is followed by luminescence lifetime measurements in water and deuterium oxide solutions, and it is found that the lanthanide ion loses at least one hydration water on binding to long double stranded DNA at pH 4.7 and pH 7. Rather different behavior is observed on binding to long or short single stranded DNA, where six water molecules are lost, independent of pH. It is suggested that in this case the lanthanide probably binds to the bases of the DNA backbone. The DNA conformation seems to be an important factor in the binding. In addition, the isotopic effect on terbium luminescence lifetime may provide a useful method to distinguish between single and double stranded DNA. DSC results are consistent with cleavage of the double helix of DNA at pH 9 in the presence of terbium. PMID- 16262312 TI - Mixed self-assembled monolayers of semirigid tetrahydro-4H-thiopyran end-capped oligo(cyclohexylidenes). AB - Single-component and mixed self-assembled monolayers (SAMs) of one- and three ring semirigid tetrahydro-4H-thiopyran end-capped oligo(cyclohexylidenes)-that is, thiopyran (1), 4-(4-cyclohexylidene-cyclohexylidene)tetrahydro-4H-thiopyran (2), and 4-(tetrahydro-4H-thiopyran-4-cyclohexylidene-4'-ylidene)tetrahydro-4H thiopyran (3)--on Au(111) substrates have been prepared and studied by cyclic voltammetry (CV), atomic force microscopy (AFM), and scanning tunneling microscopy (STM). It was found that the shortest adsorbate 1 more readily forms a SAM than 2 or 3. Notwithstanding, the SAMs of 2 or 3 are thermodynamically more stable due to favorable intermolecular attractions. Holes were made with the AFM tip establishing tilt angles of 30-50 degrees with respect to the surface normal for all SAMs. STM imaging showed well-ordered, line-shaped packing patterns with molecular resolution for the SAM of 2. Similar patterned structures were not observed for 1 and 3. Mixed SAMs were prepared by exposing a SAM of 1 to ethanol solutions of either 2 or 3. STM imaging revealed that domains of molecules of 2 or 3 amidst a monolayer of 1 are formed in both cases. Whereas in the mixed SAM of 1 and 2 the domains are irregularly shaped, circular islands of uniform size are found in the mixed SAM of 1 and 3. PMID- 16262313 TI - Surface-enhanced Raman scattering on nanoshells with tunable surface plasmon resonance. AB - Fabrication, characterization, and optical enhancement applications of bimetallic AgAu nanoparticles and nanoshells are reported. Nanoparticles with tunable surface plasmon resonances are synthesized at room temperature and characterized by transmission electron microscopy, X-ray photoelectron spectroscopy, and photon correlation spectroscopy. The collective electron oscillation of the nanoparticles shows a controllable tunability in the 400-990 nm spectral range, in agreement with plasmon absorption calculated using Mie theory, providing an optimum substrate for surface plasmon-assisted enhanced spectroscopy. Surface enhanced Raman scattering experiments show that the average enhancement factor obtained with nanoshells could be higher than those obtained with silver sols. PMID- 16262314 TI - Microscale plasma-initiated patterning (muPIP). AB - A novel technique to create biomolecular micropatterns of varying complexity on several types of polymer substrates is presented. This method uses a patterned PDMS stamp to preferentially expose or protect areas of an underlying polymer substrate from oxygen plasma. Following plasma treatment, the substrate is immersed in a biomolecular ink, whereby molecules preferentially adsorb to either the plasma-exposed or plasma-protected substrate regions, depending on the particular substrate/ink combination. Using this method, polyethylene (PE), polystyrene (PS), poly(methyl methacrylate) (PMMA), poly(dimethylsiloxane) (PDMS), and poly(hydroxybutyrate/hydroxyvalerate) (PHBV) were micropatterned with different aqueous-based biomolecular inks (i.e., goat anti-rabbit immunoglobulin G, poly-l-lysine, and bovine serum albumin (BSA)). Water contact angle measurements performed on substrates after oxygen plasma exposure showed that the hydrophilicity of substrate areas exposed to plasma was significantly greater than that of areas protected from plasma by the PDMS stamp. In addition, scanning electron microscopy results demonstrated that substrate areas exposed to plasma were physically modified (e.g., roughened) compared to adjacent, protected areas. Areas in contact with a patterned PDMS stamp during plasma exposure were found to be physically unaffected by plasma treatment, and exhibited spatial features/dimensions consistent with the corresponding features of the patterned stamp. Last, protein patterns of BSA on the polymer substrates were stable and distinct after 4 weeks of incubation at 37 degrees C. PMID- 16262315 TI - Nature of capillary condensation and evaporation processes in ordered porous materials. AB - We report on a detailed experimental study of capillary condensation-evaporation processes of N(2) in ordered mesoporous SBA-15 silica. We have carried out measurements of boundary hysteresis loops, reversal curves, and subloops in order to test whether this material behaves as an assembly of independent cylindrical pores open at both ends. With these data, we come to the conclusion that, whereas the boundary hysteresis loop has the classical shape of type H1 associated with condensation-evaporation in cylindrical pores open at both ends, the capillary evaporation does not take place at equilibrium as it is generally assumed. Moreover, the pores do not desorb independently of one another. PMID- 16262316 TI - Formation of tetra(ethylene oxide) terminated Si-C linked monolayers and their derivatization with glycine: an example of a generic strategy for the immobilization of biomolecules on silicon. AB - Surface modification with oligo(ethylene oxide) functionalized monolayers terminated with reactive headgroups constitutes a powerful strategy to provide specific coupling of biomolecules with simultaneous protection from nonspecific adsorption on surfaces for the preparation of biorecognition interfaces. To date, oligo(ethylene oxide) functionalized monolayer-forming molecules which can be activated for attachment of biomolecules but which can selectively form monolayers onto hydrogen terminated silicon have yet to be developed. Here, self assembled monolayers (SAMs) containing tetra(ethylene oxide) moieties protected with tert-butyl dimethylsilyl groups were formed by thermal hydrosilylation of alkenes with single-crystal Si(111)-H. The protection group was used to avoid side reactions with the hydride terminated silicon surface. Monolayer formation was carried out using solutions of the alkene in the high-boiling-point solvent 1,3,5-triethylbenzene. The protecting group was removed under very mild acidic conditions to yield a free hydroxyl functionality, a convenient surface moiety for coupling of biological entities via carbamate bond formation. The chemical composition and structure of the monolayers before and after deprotection were characterized by X-ray photoelectron spectroscopy (XPS) and X-ray reflectometry. To demonstrate the utility of this surface for covalent modification, two reagents were compared and contrasted for their ability to activate the surface hydroxyl groups for coupling of free amines, carbonyl diimidazole (CDI), and disuccinimidyl carbonate (DSC). Analysis of XP spectra before and after activation by CDI or DSC, and after subsequent reaction with glycine, provided quantitative information on the extent of activation and overall coupling efficiencies. CDI activated surfaces gave poor coupling yields under various conditions, whereas DSC mediated activation followed by aminolysis at neutral pH was found to be an efficient method for the immobilization of amines on tetra(ethylene oxide) modified surfaces. PMID- 16262317 TI - Improvement of the Derjaguin-Broekhoff-de Boer theory for the capillary condensation/evaporation of nitrogen in spherical cavities and its application for the pore size analysis of silicas with ordered cagelike mesopores. AB - In a previous work, we proposed an improvement of the Derjaguin-Broekhoff-de Boer (DBdB) theory for capillary condensation/evaporation in open-ended cylindrical mesopores. In this paper, we report a further extension of this approach to the capillary condensation/evaporation of nitrogen in siliceous spherical cavities. The main idea of this improvement is to employ the Gibbs-Tolman-Koenig-Buff equation to predict the variation of the surface tension in spherical mesopores. In addition, the statistical film thickness (the so-called t-curve), which is evaluated accurately on the basis of adsorption isotherms measured for MCM-41 materials, is used instead of the originally proposed t-curve to take into account the excess chemical potential due to the surface forces. It is shown that the aforementioned modifications of the original DBdB theory that was refined by Ravikovitch and Neimark have significant implications for the pore size analysis of cagelike mesoporous silicas. To verify the proposed improvement of the DBdB pore size analysis (IDBdB), two series of FDU-1 samples, which are well-defined cagelike mesoporous materials (composed of siliceous spherical cavities interconnected by short necks), were used for the evaluation of the pore size distributions (PSDs). The correlation between the spinodal condensation point in the spherical pores predicted by the nonlocal density functional theory (NDFT) developed by Ravikovitch and Neimark and that predicted by the IDBdB theory is very good in the whole range of mesopores. This feature is mirrored to the realistic PSD characterized by the bimodal structure of pores computed from the IDBdB theory. As in the case of open-ended cylindrical pores, the improvement of the classical DBdB theory preserves its simplicity and simultaneously ensures a significant improvement of the pore size analysis, which is confirmed by the independent estimation of the average pore size by the NDFT and the powder X-ray diffraction method. PMID- 16262318 TI - Simple methods for the direct assembly, functionalization, and patterning of acid terminated monolayers on Si111. AB - This article describes mild methods to directly assemble, functionalize, and pattern monolayers of undecylenic acid on hydrogen-terminated Si(111). These monolayers were assembled under very mild conditions from a neat solution of undecylenic acid containing 0.1 mol % 4-(decanoate)-2,2,6,6 tetramethylpiperidinooxy at room temperature without the need for UV light. Because of these mild conditions, monolayers exposing carboxylic acids could be assembled in one step without the need to protect the acid prior to its assembly. The monolayers were extensively characterized by horizontal attenuated total reflection infrared spectroscopy, X-ray photoelectron spectroscopy (XPS), and contact angle goniometry. The monolayers bonded to the silicon surface preferentially through the olefin with no detectable bonds between the carboxylic acids and silicon. The crystallinity of the monolayer was studied by infrared spectroscopy through the antisymmetric--v(a)(CH(2))--and symmetric--v(s)(CH(2))- stretches for methylene. Because it is important for future applications to assemble functional surfaces, methods to react the acid-terminated monolayers with trifluoroacetic anhydride and triethylamine to yield a symmetric anhydride on the monolayer were studied. These anhydrides were reacted with a variety of milligram-quantity amines to yield amide-terminated surfaces. This method was general, and a variety of amines could be bonded to the monolayer. The stabilities of these monolayers upon exposure to ambient conditions and under a variety of solvents were described. Because patterned monolayers have found wide applications, we have developed methods to pattern 1-octadecylamine and poly(ethylenimine) on the micrometer scale using soft lithography. In addition, polymer brushes of polynorbornene with thicknesses from 32 to 150 nm were grown from monolayers patterned with the Grubbs' catalyst. The patterned surfaces were imaged by scanning electron microscopy, scanning probe microscopy, and ellipsometry to determine the thicknesses of the patterns and the fidelity of the method. PMID- 16262319 TI - FTIR spectroscopic study of titanium-containing mesoporous silicate materials. AB - The surface acidity of different mesoporous titanium-silicates, such as well organized hexagonally packed Ti-MMM, Ti-MMM-2, Ti-SBA-15, and amorphous TiO(2) SiO(2) mixed oxides (aerogels and xerogels), was studied by means of FTIR spectroscopy of CO adsorbed at 80 K and CD(3)CN adsorbed at 293 K. The surface hydroxyl groups of mesoporous titanium-silicates with 2-7 wt % Ti revealed a Bronsted acidity slightly higher to that of pure silicate. TiO(2)-SiO(2) xerogels revealed the highest Bronsted acidity among the titanium-silicates studied. CO adsorption revealed two additional sites on the surface in comparison to pure silicate, characterized by nu(CO) from 2185 (high pressure) to 2178 (low pressure) cm(-1) and from 2174 (high pressure) to 2170 (low pressure) cm(-1). These bands are due to CO adsorbed on isolated titanium cations in the silica surrounding or having one Ti(4+) cation in their second coordination sphere and due to CO interactions with Ti-OH groups, respectively. CD(3)CN adsorption similarly revealed the existence of two additional sites, which were not detected for pure silicate: at 2289 cm(-1) due to CD(3)CN interaction with titanol groups and from 2306 (low pressure) to 2300 (high pressure) cm(-1) due to acetonitrile interaction with isolated framework titanium cations with probably one Ti(4+) cation in their second coordination shell. The spectroscopic results are compared with computational data obtained on cluster models of titanium-silicate with different titanium content. According to the IR data, the Ti accessibility on the surfaces for mesoporous titanium-silicates with similar Ti loading (2 wt %) was found to fall in the order TiO(2)-SiO(2) aerogel approximately TiO(2)-SiO(2) xerogel > Ti-MMM approximately Ti-MMM-2 > Ti-SBA-15. This order (except TiO(2) SiO(2) xerogel) correlates with the catalytic activity found previously for titanium-silicates in 2,3,6-trimethylphenol oxidation with H(2)O(2). PMID- 16262320 TI - Sorbitan tristearate layers at the air/water interface studied by shear and dilatational interfacial rheology. AB - The shear and dilatational rheology of condensed interfacial layers of the water insoluble surfactant sorbitan tristearate at the air/water interface is investigated. A new interfacial shear rheometer allows measurements in both stress- and strain-controlled modes, providing comprehensive interfacial rheological information such as the interfacial dynamic shear moduli, the creep response to a stress pulse, the stress relaxation response to a strain step, or steady shear curves. Our experiments show that the interfacial films are both viscoelastic and brittle in nature and subject to fracture at small deformations, as was supported by in-situ Brewster angle microscopy performed during the rheological experiments. Although any large-deformation test is destructive to the sample, it is still possible to study the linear viscoelastic regime if the deformations involved are controlled carefully. Complementary results for the dilatational rheology in area step compression/expansion experiments are reported. The dilatational behavior is predominantly elastic throughout the frequency spectrum measured, whereas the layers exhibit generalized Maxwell behavior in shear mode within a deformation frequency regime as narrow as two decades, indicating the presence of additional relaxation mechanisms in shear as opposed to expansion/compression. If the transient rheological response from stress relaxation experiments is considered, then the data can be described well with a stretched exponential model both in the shear and dilatational deformations. PMID- 16262321 TI - Effect of Mn and Fe on the reactivity of sulfated zirconia toward H2 and n butane: a diffuse reflectance IR spectroscopic investigation. AB - Sulfated zirconia (SZ) and sulfated zirconia promoted with 2 wt % manganese (MnSZ) or iron (FeSZ), all active in n-butane isomerization, were investigated using diffuse reflectance Fourier transform IR spectroscopy (DRIFTS). By adsorption of H(2) at 77 K or of n-butane at room temperature, it was found that the promoters neither enhance the Lewis nor the Bronsted acid strength. SZ and promoted SZ do not exhibit higher acid strength than zeolites. In a batch experiment using 70 hPa of H(2), SZ did not react at 473 K. Reaction of H(2) with MnSZ produced water (band at 5242 cm(-1)) and a decrease in the sulfate groups (multiple bands). Heating of SZ in 10 hPa n-butane to 573 K caused total reduction of sulfate to H(2)S (2583, 2570 cm(-1)) and partial and total oxidation of butane to olefinic species (3062 cm(-1)), CO(2), and water. MnSZ and FeSZ reacted with n-butane already at 373 K; products of skeletal isomerization (methyne CH vibration at 2910 cm(-1)) were detected and sulfate groups were consumed. Rather than increasing the acidity, the promoters enhance the oxidation potential of sulfate and facilitate alkane activation via oxidative dehydrogenation. PMID- 16262322 TI - A reversibly switching block copolymer surface. AB - We present linear (AB)(n)() multiblock copolymers that exhibit a thermally induced reversible alteration of the surface composition at a sharply defined transition temperature T(s) of 120-170 degrees C depending on the polymer structure. At temperatures below T(s) the surface consists of block A, a 4,4' methylenediphenyl diisocyanate (4,4'-MDI) type polyurea, whereas above T(s) the hydrophobic block B, a poly(ricinoleic acid hexanediol ester) dominates the surface composition. The ratio of surface concentrations c(A)/c(B) changes by a factor of at least 1000 within an analyzed depth of approximately 10 A. The full A-B surface transition is obtained within minutes. A mechanism is proposed where microphase crystallization of block A in the bulk effectively locks surface segregation of the hydrophobic block B, yielding an A-rich surface. The topology of the copolymers imposes sufficient restrictions for the lateral separation of the connected constituents such that surface segregation is largely reduced. Only above the transition temperature T(s) of microphase crystallization of block A can block B segregate to the surface, yielding a B-rich surface. Such a scheme of competing self-organizing processes in copolymers may potentially be used to reversibly switch surface properties such as adhesion and wetting in various applications. PMID- 16262323 TI - Phase separation of ternary self-assembled monolayers into hydrophobic 1 dodecanethiol domains and electrostatically stabilized hydrophilic domains composed of 2-aminoethanethiol and 2-mercaptoethanesulfonic acid on Au(111). AB - Ternary self-assembled monolayers (SAM) composed of 2-aminoethanethiol (AET), 2 mercaptoethanesulfonic acid (MES), and 1-dodecanethiol (DDeT) form two types of domains as if it were a two-component SAM: DDeT-rich hydrophobic domains and electrostatically stabilized hydrophilic domains composed of MES and AET on Au(111). MES and AET behave virtually as a single surface-active species. Two distinct reductive desorption peaks in cyclic voltammograms (CV) and binarized images of scanning tunneling microscopy clearly show nanometer scale, yet macroscopically distinguishable, phase separation over a wide range of the mixing ratio of DDeT and MES-AET in the bathing solution. X-ray photoelectron spectroscopy measurements indicate that the ratio of MES to AET in the hydrophilic domains is unity and that both terminal groups are in the charged states, that is, the sulfonate group and the ammonium group. With decreasing the total concentration of the thiols, the mole fraction of DDeT in the bathing solution at which the surface coverage of MES-AET domains is equal to that of DDeT domains dramatically decreases. This suggests that the adsorption kinetics plays a crucial role in the formation of the domains structure. PMID- 16262324 TI - High-flux nanofiltration membranes prepared by adsorption of multilayer polyelectrolyte membranes on polymeric supports. AB - Layer-by-layer deposition of anionic and cationic polyelectrolytes readily converts polymeric ultrafiltration membranes into materials capable of nanofiltration. ATR-FTIR spectra confirm that layer-by-layer deposition occurs on the ultrafiltration substrates, and adsorption of as few as 2.5 bilayers of poly(styrenesulfonate) (PSS)/protonated poly(allylamine) (PAH) or 3.5 bilayers of PSS/poly(diallyldimethylammonium chloride) (PDADMAC) reduces the molecular weight cutoff of polyethersulfone ultrafiltration supports from 50 kDa to <500 Da. Deposition of multilayer polyelectrolyte films on 300 and 500 kDa membranes also decreases molecular weight cutoffs, but solute rejections are significantly lower when using these supports, suggesting that the polyelectrolyte films do not completely cover large (0.2-0.4 microm in diameter) pores. On the 50 kDa substrates, PSS/PDADMAC films containing 3.5 bilayers exhibit a 95% rejection of SO(4)(2-) and a chloride/sulfate selectivity of 27, whereas 4.5-bilayer PSS/PAH coatings show a glucose/raffinose selectivity of 100. Pure water flux for [PSS/PAH](3)PSS-coated membranes at 4.8 bar is 1.6 m(3)/(m(2)day), which is more than 2-fold higher than that through a commercial 500 Da membrane. PMID- 16262325 TI - Effect of humidity on the adsorption kinetics of lung surfactant at air-water interfaces. AB - The in vitro adsorption kinetics of lung surfactant at air-water interfaces is affected by both the composition of the surfactant preparations and the conditions under which the assessment is conducted. Relevant experimental conditions are surfactant concentration, temperature, subphase pH, electrolyte concentration, humidity, and gas composition of the atmosphere exposed to the interface. The effect of humidity on the adsorption kinetics of a therapeutic lung surfactant preparation, bovine lipid extract surfactant (BLES), was studied by measuring the dynamic surface tension (DST). Axisymmetric drop shape analysis (ADSA) was used in conjunction with three different experimental methodologies, i.e., captive bubble (CB), pendant drop (PD), and constrained sessile drop (CSD), to measure the DST. The experimental results obtained from these three methodologies show that for 100% relative humidity (RH) at 37 degrees C the rate of adsorption of BLES at an air-water interface is substantially slower than for low humidity. It is also found that there is a difference in the rate of surface tension decrease measured from the PD and CB/CSD methods. These experimental results agree well with an adsorption model that considers the combined effects of entropic force, electrostatic interaction, and gravity. These findings have implications for the development and evaluation of new formulations for surfactant replacement therapy. PMID- 16262326 TI - Enhanced photodegradation of 2,4,6-trichlorophenol over palladium phthalocyaninesulfonate modified organobentonite. AB - This article explores a supported photocatalyst of palladium(II) phthalocyaninesulfonate (PdPcS) onto organoclay for removal of 2,4,6 trichlorophenol (TCP) from water under visible light (lambda > or = 450 nm) irradiation. The composite clay was not only a good sorbent for uptake of TCP from water, but it also exhibited notable activity for TCP oxidation via singlet molecular oxygen, generated in situ from PdPcS photosensitization. Complete dechlorination of TCP could be achieved after TCP was totally oxidized. The initial rate of TCP degradation was observed to increase with the initial amount of TCP sorption, the kinetics following well the Langmuir-Hinshelwood equation. The resulting rate constant of TCP oxidation was found to increase with the alkyl chain length of the intercalated surfactant from dodecyltrimethylammonium to cetyltrimethylammonium and to octadecyltrimethylammonium. Such the trend in photoactivity was in agreement with the catalyst capacity for TCP sorption from aqueous medium. The result showed that enrichment of toxic TCP on the catalyst was indeed an efficient way for enhancement of the photosensitized degradation of target pollutant. However, PdPcS loading led to a notable decrease in TCP sorption, because of partial blocking of the sorptive sites. The optimal loading of PdPcS was about 1.0 wt %. The catalyst was characterized by nitrogen adsorption, X-ray diffraction, visible diffuse reflectance spectroscopy, and TCP sorption, which revealed that the incorporated PdPcS, mainly in the form of photoactive monomer, coexisted with the intercalated surfactant in the bentonite interlayers. Seven repeated experiments demonstrated that the composite clay was relatively stable and could be repeatedly used for sorption and degradation of TCP pollutant via molecular oxygen and visible light. PMID- 16262327 TI - Interactions of benzoic acid and phosphates with iron oxide colloids using chemical force titration. AB - Colloidal iron oxides are an important component in soil systems and in water treatment processes. Humic-based organic compounds, containing both phenol and benzoate functional groups, are often present in these systems and compete strongly with phosphate species for binding sites on the iron oxide surfaces. Here, we examine the interaction of benzoate and phenolic groups with various iron oxide colloids using atomic force microscopy (AFM) chemical force titration measurements. Self-assembled monolayers (SAMs) of 4-(12 mercaptododecyloxy)benzoic acid and 4-(12-mercaptododecyloxy)phenol were used to prepare chemically modified Au-coated AFM tips, and these were used to probe the surface chemistry of a series of iron oxide colloids. The SAMs formed were also characterized using scanning tunneling microscopy, reflection-absorption infrared spectroscopy, and X-ray photoelectron spectroscopy. The surface pK(a) of 4-(12- mercaptododecyloxy)benzoic acid has been determined to be 4.0 +/- 0.5, and the interaction between the tip and the sample coated with a SAM of this species is dominated by hydrogen bonding. The chemical force titraton profile for an AFM probe coated with 4-(12- mercaptododecyloxy)benzoic acid and a bare iron oxide colloid demonstrates that the benzoic acid function group interacts with all three types of iron oxide sites present on the colloid surface over a wide pH range. Similar experiments were carried out on colloids precipitated in the presence of phosphoric, gallic, and tannic acids. The results are discussed in the context of the competitive binding interactions of solution species present in soils or in water treatment processes. PMID- 16262328 TI - Enhanced anion electroadsorption into carbon molecular sieve electrodes in acidic media. AB - We previously showed that, for neutral electrolytes of small cations and relatively larger anions, it is possible to design certain pore sizes in active carbons that are large enough to electroadsorb cations but too small to allow anion electroadsorption. This situation leads to an electrical double-layer (EDL) capacitance that is significant only at potentials that are negative to the potential of zero charge (PZC); hence, much smaller capacitance is measured at potentials positive to the PZC. It was found that when the electrolyte is a strong acid (e.g., H(2)SO(4), HCl), a considerable capacitance is observed at positive potentials, even when the average pore size is too small to allow the insertion of large anions in neutral electrolyte solutions. This effect disappears when the pore size becomes considerably larger than the size of the ions. In this case, the EDL capacitance at positive potentials for both neutral and acidic solutions is comparable. The following four-step mechanism was found to comply best with the experimental data: (1) By acid catalysis, the protons form carbonium species within the conjugated carbon network. (2) The anions react with the carbonium ions, providing uncharged species in an activated state, which are chemibound as surface groups to the walls of the pores. (3) Because these surface groups are effectively much smaller in size than are the charged ions, they can migrate by chemical bond exchange within the carbon skeleton via constrictions (known to exist in microporous and molecular sieving carbons), which are too narrow to accommodate hydrated charged species. (4) Upon reaching wider spaces, the uncharged species are reionized and solvated by water molecules, which can fill small pores. The justification for the above mechanism is thoroughly discussed and demonstrated by the experimental results. PMID- 16262329 TI - Electron transfer in metal-molecule-metal junction composed of self-assembled monolayers of helical peptides carrying redox-active ferrocene units. AB - Electronic properties of three kinds of helical peptides with or without redox active ferrocene units were investigated by using scanning tunneling microscopy under ultrahigh vacuum. The currents through the helical peptides carrying ferrocene units at the molecular terminals became significantly larger than that through a reference peptide without any ferrocene units. On the other hand, ferrocene units in the middle of the peptide chain did not affect the current voltage characteristics. These results indicate that the ferrocene units near the metal electrode should play an important role for efficient electron transfer between the metal and the peptide molecules, which process is one of the rate determining steps for characterizing molecular conductance in metal-molecule metal junction. PMID- 16262330 TI - New reverse micelle surfactant systems optimized for high-resolution NMR spectroscopy of encapsulated proteins. AB - Sodium bis(2-ethylhexyl)sulfosuccinate (AOT) is a surfactant commonly used to encapsulate water soluble proteins within the aqueous core of a reverse micelle. In the context of high-resolution NMR studies of encapsulated proteins the size of the resulting reverse micelle is critically important. We have designed and synthesized a short AOT analogue, 3,3-dimethyl-1-butylsulfosuccinate sodium salt and determined that it is able to form reverse micelles and to encapsulate the protein ubiquitin with high structural fidelity. AOT is often found to significantly destabilize encapsulated proteins, largely through charge-charge interactions between the anionic headgroup and the surface of the protein. Here we demonstrate, for the first time, that proportional mixtures of anionic and cationic surfactants can form reverse micelles that are also capable of protein encapsulation with high fidelity. PMID- 16262331 TI - Cobalt and magnesium ferrite nanoparticles: preparation using liquid foams as templates and their magnetic characteristics. AB - An easy and convenient method for the synthesis of cobalt and magnesium ferrite nanoparticles is demonstrated using liquid foams as templates. The foam is formed from an aqueous mixture of an anionic surfactant and the desired metal ions, where the metal ions are electrostatically entrapped by the surfactant at the thin borders between the foam bubbles and their junctions. The hydrolysis is carried out using alkali resulting in the formation of desired nanoparticles, with the foam playing the role of a template. However, in the formation of ferrites with the formula MFe(2)O(4), where the metal ion and iron possess oxidation states of +2 and +3, respectively, forming a foam from a 1:2 mixture of the desired ionic solutions would lead to a foam composition at variance with the original solution mixture because of greater electrostatic binding of ions possessing a greater charge with the surfactant. In our procedure, we circumvent this problem by preparing the foam from a 1:2 mixture of M(2+) and Fe(2+) ions and then utilizing the in situ conversion of Fe(2+) to Fe(3+) under basic conditions inside the foam matrix to get the desired composition of the metal ions with the required oxidation states. The fact that we could prepare both CoFe(2)O(4) and MgFe(2)O(4) particles shows the vast scope of this method for making even multicomponent oxides. The magnetic nanoparticles thus obtained exhibit a good crystalline nature and are characterized by superparamagnetic properties. The magnetic features observed for CoFe(2)O(4) and MgFe(2)O(4) nanoparticles are well in accordance with the expected behaviors, with CoFe(2)O(4) particles showing higher blocking temperatures and larger coercivities. These features can easily be explained by the contribution of Co(2+) sites to the magnetocrystalline anisotropy and the absence of the same from the Mg(2+) ions. PMID- 16262332 TI - Biocompatibility of gold nanoparticles and their endocytotic fate inside the cellular compartment: a microscopic overview. AB - Macrophages are one of the principal immune effector cells that play essential roles as secretory, phagocytic, and antigen-presenting cells in the immune system. In this study, we address the issue of cytotoxicity and immunogenic effects of gold nanoparticles on RAW264.7 macrophage cells. The cytotoxicity of gold nanoparticles has been correlated with a detailed study of their endocytotic uptake using various microscopy tools such as atomic force microscopy (AFM), confocal-laser-scanning microscopy (CFLSM), and transmission electron microscopy (TEM). Our findings suggest that Au(0) nanoparticles are not cytotoxic, reduce the production of reactive oxygen and nitrite species, and do not elicit secretion of proinflammatory cytokines TNF-alpha and IL1-beta, making them suitable candidates for nanomedicine. AFM measurements suggest that gold nanoparticles are internalized inside the cell via a mechanism involving pinocytosis, while CFLSM and TEM studies indicate their internalization in lysosomal bodies arranged in perinuclear fashion. Our studies thus underline the noncytotoxic, nonimmunogenic, and biocompatible properties of gold nanoparticles with the potential for application in nanoimmunology, nanomedicine, and nanobiotechnology. PMID- 16262333 TI - Effect of a metal alloy fuel catalyst on bacterial growth. AB - Many microorganisms have been demonstrated to utilize petroleum fuel products to fulfill their nutritional requirement for carbon. As a result, the ability of these microbes to degrade fuel has both a deleterious affect as well as beneficial applications. This study focused on the undesired ability of bacteria to grow on fuel and the potential for some metal alloys to inhibit this biodegradation. The objective of this study was to review the pattern of growth of two reference strains of petroleum-degrading bacteria, Pseudomonas oleovorans and Rhodococcus rhodocrous, in a specific hydrocarbon environment in the presence of a commercially available alloy. The alloy formulated and supplied by Advanced Power Systems International Inc. (APSI) is sold for fuel reformulation and other purposes. The components of the alloy used in the study were antimony, tin, lead, and mercury formulated as pellets. Surface characterization also showed the presence of tin oxide and lead amalgam phases. Hydrocarbon used for the study was primarily 87-octane gasoline. The growth of the bacteria in the water and mineral supplemented gasoline mixture over 6-8 weeks was monitored by the viable plate count method. While an initial increase in bacteria occurred in the first week, overall bacterial growth was found to be suppressed in the presence of the alloy. Results also indicate that the alloy surface characteristics that convey the catalytic activity may also contribute to the observed antibacterial activity. PMID- 16262334 TI - Interactions of apo cytochrome C with alternating copolymers of maleic acid and alkene. AB - Apo cytochrome c (apo cyt c) tends to aggregate at alkali pH. Poly(isobutylene alt-maleic acid) (PIMA) is soluble molecularly, whereas poly(1-tetradecene-alt maleic acid) (PTMA) forms particles that tend to dissociate by increasing pH and decreasing concentration. Dynamic light scattering and surface plasmon resonance are used to investigate the interactions of PIMA and PTMA with apo cyt c at different pH values to understand the mechanism of the interactions. When the positive or negative charges are in excess, the copolymer-protein complex particles can be stabilized by the charges on the surface. When the ratio of the positive to negative charges is close to the stoichiometric value, precipitation occurs. At pH 11.8, both PTMA and apo cyt c carry negative charges, but the hydrophobic interaction makes them form complexes. A competition exists between the interaction of the copolymer with apo cyt c and the self-aggregation of PTMA or apo cyt c alone. The interaction of PIMA or PTMA with apo cyt c at neutral and alkali pH destroys the aggregation of PTMA or apo cyt c and forms new complex particles. PMID- 16262335 TI - Langmuir monolayer as a tool toward visualization of a specific DNA-protein complex. AB - Immobilization and imaging of protein molecules and protein-DNA complexes on a Langmuir-Blodgett (LB) substrate have been explored here. We have prepared a nickel-arachidate (NiA) monolayer and characterized it through pressure-area isotherm on a LB trough. Recombinant RNA polymerase from Escherichia coli, where the largest subunit was replaced with the same gene having a series of histidine amino acids at the C-terminus end of the protein, was immobilized over the Ni arachidate monolayer through a Ni(II)-histidine interaction. A single molecule of RNA polymerase could be seen through intermittent-contact atomic force microscopy (AFM). Under the condition of the formation of the LB monolayer, the enzyme molecules were arrayed and transcriptionally active. Interestingly, they could pick up sequence specific DNA molecules from the subphase in an oriented fashion. On the other hand, preformed RNA polymerase Ni(II)-arachidate monolayers bound DNA haphazardly when no surface pressure was employed. PMID- 16262336 TI - Using nile red-adsorbed gold nanoparticles to locate glutathione within erythrocytes. AB - An aqueous solution of Nile Red (NR)-absorbed 32-nm gold nanoparticles (AuNPs) have been used to sense glutathione (GSH). When the NR product is displaced by GSH on the AuNP surface, the fluorescence of the solution increases and the AuNPs aggregate. To determine the concentration and distribution of GSH within erythrocyte cells, a homemade fluorescence and scattering microscope was constructed. This system allows monitoring, within individual cells, of the uptake and transportation of the NRAuNPs and the displacement of the NR product from the NRAuNP surface by GSH. The fluorescence and scattering images clearly indicate the location of GSH inside the cells; these findings are supported by images recorded using 2,3-naphthalenedicarboxaldehyde, which is a highly selective fluorogenic reagent for GSH. Microscopic fluorescence measurements of the NRAuNPs revealed that the GSH concentration inside erythrocyte cells is 1.30 +/- 0.31 mM. To confirm this result, lysed erythrocyte cells were analyzed by applying capillary electrophoresis in conjunction with laser-induced fluorescence using NRAuNPs; accordingly, the average GSH concentration in a single erythrocyte cell was determined to be 1.32 +/- 0.06 mM. PMID- 16262337 TI - Resolution of the vertical and horizontal heterogeneity of adsorbed collagen layers by combination of QCM-D and AFM. AB - Collagen (type I from calf skin) adsorption on polystyrene (PS) and plasma oxidized polystyrene (PSox) was studied, using a quartz crystal microbalance with energy dissipation measurements (QCM-D) and atomic force microscopy (AFM) in tapping mode. Radio-labeled collagen was used to measure the adsorbed amount and the ability of adsorbed collagen to exchange with molecules in the solution. The results show that the collagen adlayer consists of two parts: a dense and thin sheet in which fibrils are formed (directly observed by AFM) and an overlying thick layer (up to 200 nm) containing protruding molecules or bundles which are in very low concentration but modify noticeably the local viscosity. The thickness and viscosity of the semi-liquid adlayer both increase with adsorption time and collagen concentration. Fibril formation near the surface also increases with time and collagen concentration and occurs more readily on PS compared to PSox. Radiochemical measurements show that this may be related to the larger mobility of molecules adsorbed on PS, presumably owing to a smaller number of binding points. PMID- 16262338 TI - Neuronal activation by GPI-linked neuroligin-1 displayed in synthetic lipid bilayer membranes. AB - We have characterized, in vitro, interactions between hippocampal neuronal cells and silica microbeads coated with synthetic, fluid, lipid bilayer membranes containing the glycosylphosphatidyl inositol (GPI)-linked extracellular domain of the postsynaptic membrane protein neuroligin-1. These bilayer-neuroligin-1 beads activated neuronal cells to form presynaptic nerve terminals at the point of contact in a manner similar to that observed for live PC12 cells, ectopically expressing the full length neuroligin-1. The synthetic membranes exhibited biological activity at neuroligin-1 densities of approximately 1 to 6 proteins/microm(2). Polyolycarbonate beads with neuroligin-1 covalently attached to the surface failed to activate neurons despite the fact that neuroligin-1 binding activity is preserved. This implies that a lipid membrane environment is likely to be essential for neuroligin-1 activity. This technique allows the study of isolated proteins in an environment that has physical properties resembling those of a cell surface; proteins can diffuse freely within the membrane, retain their in vivo orientations, and are in a nondenatured state. In addition, the synthetic membrane environment affords control over both lipid and protein composition. This technology is easily implemented and can be applied to a wide variety of cellular studies. PMID- 16262339 TI - Gold nanoparticle decoration of DNA on silicon. AB - Electrostatic assembly of cationic nanoparticles onto the negatively charged backbone of double-stranded DNA has been shown to produce one-dimensional chains with potential use as nanoelectronic components. In this paper, micron long DNA templates stretched on aminosilane- and hexamethyldisilazane-modified silicon surfaces are used to assemble 3.5 nm gold nanoparticles passivated with cationic thiocholine. Atomic force microscopy is used to analyze the density and defects along the approximately 5 nm high structures, with comparison between positively charged and neutral surfaces. Low background adsorption of nanoparticles is facilitated by both these surface chemistries, while the neutral surface yields a more densely packed assembly. PMID- 16262340 TI - Porous and electrically conductive polypyrrole-poly(vinyl alcohol) composite and its applications as a biomaterial. AB - Bulk modification of polypyrrole (PPY) with poly(vinyl alcohol) (PVA) was carried out by the electropolymerization of pyrrole in the presence of PVA in the reaction solution, with tetraethylammonium perchlorate (TEAP) as the electrolyte. The surface morphology of the as-synthesized PPY-TEAP-PVA film was investigated using scanning electron microscopy, and the film was further characterized using X-ray photoelectron spectroscopy, electrical conductivity, the water contact angle, and BET surface area measurements. The PPY-TEAP-PVA composite is electrically conductive, hydrophilic, and microporous with a high surface area. Its potential as a biomaterial was investigated with respect to its blood compatibility and function as a substrate for biosensor fabrication and cell culture. The presence of PVA in the film attenuates blood protein adsorption, and the porous nature of the PPY-TEAP-PVA film results in a 10-fold increase in the amount of glucose oxidase covalently immobilized on the film over that on a nonporous PPY film. PC12 cell attachment and growth on the PPY-TEAP-PVA film was also shown to be enhanced compared with that on tissue culture polystyrene. The attached cells proliferated and formed a monolayer on the film surface after 48 h of seeding. PMID- 16262341 TI - Bacillus atrophaeus outer spore coat assembly and ultrastructure. AB - Our previous atomic force microscopy (AFM) studies successfully visualized native Bacillus atrophaeus spore coat ultrastructure and surface morphology. We have shown that the outer spore coat surface is formed by a crystalline array of approximately 11 nm thick rodlets, having a periodicity of approximately 8 nm. We present here further AFM ultrastructural investigations of air-dried and fully hydrated spore surface architecture. In the rodlet layer planar and point defects as well as domain boundaries similar to those described for inorganic and macromolecular crystals were identified. For several Bacillus species rodlet structure assembly and architectural variation appear to be a consequence of species-specific nucleation and crystallization mechanisms that regulate the formation of the outer spore coat. We propose a unifying mechanism for nucleation and self-assembly of this crystalline layer on the outer spore coat surface. PMID- 16262342 TI - Novel sequence for generating glycopolymer tethered on a membrane surface. AB - Cell surface carbohydrates, usually binding with other biomacromolecules (such as lipids and proteins), are involved in numerous biological functions, including cellular recognition, adhesion, cell growth regulation, and inflammation. Synthetic carbohydrate-based polymers, so-called glycopolymers, are emerging as important well-defined tools for investigating carbohydrate-based biological processes and for simulating various functions of carbohydrates. In this study, a novel two-step sequence for the generation of a glycopolymer layer tethered on a polypropylene microporous membrane is described. First, a UV-induced graft polymerization of 2-aminoethyl methacrylate hydrochloride (AEMA) was carried out on the membrane to generate an amino-functionalized surface, and the effects of polymerization factors (monomer/initiator concentration and UV irradiation time) on the grafting density were studied. Second, sugar moieties were bound with the grafted functional layer to form glycopolymer by the reaction between the amino groups on the membrane surface and carbohydrate lactones. Chemical analysis by Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy combined with surface morphology observation by scanning electron microscopy confirmed the graft polymerization of AEMA and the formation of glycopolymer. The decreases of water contact angle and protein adsorption on the membrane revealed the enhancement of hydrophilicity and protein resistance due to the typical characteristics of the glycopolymer tethered on the surface. These results indicated that the novel sequence reported in this work is a facile process to form glycopolymer-modified surfaces. PMID- 16262343 TI - Synthesis of zeolite nanocrystals at room temperature. AB - Zeolite A nanoparticles were synthesized under room-temperature conditions from a very reactive organic-template-free gel system. The optimization of the syntheses parameters, namely, the composition of the initial system and the careful choice of the reactants, allowed the crystallization to be accomplished within 3 days. At this stage the individual zeolite crystals were in the range of 100-300 nm without well-developed crystal faces. The prolongation of the synthesis time up to 10 days led to formation of larger well-faceted cubic crystals averaging about 400-500 nm in size. The high-resolution transmission electron microscopy (HRTEM) study revealed that a thin layer of amorphous material covers the zeolite particles acting as a binder between individual zeolite crystals. The postsynthesis treatment of the product in NH(3) media under ultrasonic radiation disintegrated the loosely attached zeolite particles and decreased the fraction of zeolite A particles with low colloidal stability. The employed approach, however, did not result in complete disintegration of aggregated crystals. The zeolite crystals obtained under ambient conditions were characterized by XRD, SEM, dynamic light scattering, and N(2) adsorption measurements. PMID- 16262344 TI - Endohedral condensation and higher exohedral coverage of Kr on open single-walled carbon nanotubes at 77 K. AB - An isotherm quasi-discontinuity, or a near-vertical step, at 177 microTorr indicative of Kr condensation inside open single-walled carbon nanotubes (SWNT) has been observed at 77 K. The isotherm shows double adsorption-branch structure attributed to the existence of two endohedral phases of confined Kr. Three well pronounced steps corresponding to the formation of various exohedral phases are present in the high-density (low-pressure) branch. The desorption branch exhibits three rounded steps assigned to higher order exohedral coverage. PMID- 16262345 TI - Structure of a cyclodextrin functionalized anionic clay: XRD analysis, spectroscopy, and computer simulations. AB - Carboxy-methyl beta-cyclodextrin (CMCD) cavities have been intercalated within the galleries of anionic clay, Mg-Al layered double hydroxide (LDH). The cyclodextrin functionalized LDH has been reported to adsorb neutral and nonpolar guest molecules. X-ray diffraction, IR, and Raman vibrational spectroscopy and (13)C CPMAS NMR have been used to characterize the confined CMCD molecules, whereas molecular dynamics simulations have been used to probe the interlayer arrangement and orientation of the intercalated species. Spectroscopic measurements as well as MD simulations show that there is no significant change in the geometry of the CMCD cavity on intercalation. Within the galleries of the anionic clay, the CMCD anions are arranged as bilayers with the carboxy methyl substituents, located at the narrower opening of the bucket-like cyclodextrin toroid, anchored to the LDH sheet. This arrangement leaves the wider opening of the CMCD anion facing away from the layers allowing the interior of the cyclodextrin cavity to be accessible to guest molecules. Finally, the hydrophobicity of the anchored cyclodextrin cavity has been characterized using fluorescence from pyrene included within it. PMID- 16262346 TI - Nanoparticles of calcium hydroxide for wood conservation. The deacidification of the Vasa warship. AB - The seventeenth century Swedish warship Vasa represents a unique case in the study of ancient wrecks and a challenge for finding new methods for artifacts conservation. The presence of sulfuric acid inside the wooden structure of Vasa is one of the possible causes of chemical damage of the wood. During recent investigations, pH values below 2 were observed inside the wreck in several places. Neutralization treatments temporarily raised the surface pH about 6 units, but after a few months the pH reverted back to original values. In this study we show that wood from the Vasa warship can be deacidified by using a dispersion in 2-propanol of calcium hydroxide nanoparticles. These particles can penetrate into the wood allowing a very efficient deacidification. Alkaline nanoparticles are converted into calcium sulfate without mechanical stress to the wood's lumens. Additional applications produce an excess of alkaline nanoparticles that are converted into carbonate, an alkaline reservoir to protect the wood from further acid attack. Artificial aging of Vasa wood demonstrates that nanoparticles facilitate protection of wood toward further acid degradation. PMID- 16262347 TI - Polyurethanes containing oxetane-derived poly(2,2-substituted-1,3-propylene oxide) soft blocks: copolymer effect on wetting behavior. AB - Hydroxy-terminated poly(2,2-substituted-1,3-propylene oxide) telechelics and co telechelics bearing semifluorinated (R = -CH(2)OCH(2)(CF(2))(n)CF(3), n = 0, 1) and/or bromomethyl pendant groups were synthesized from the corresponding 3,3 substituted oxetanes. The new telechelics were incorporated in polyurethanes (PUs) with isophorone diisocyanate (IPDI) and 1,4-butanediol (BD) as the hard block. Surface properties were evaluated using tapping mode atomic force microscopy (TM-AFM) and dynamic contact angle (DCA) analysis. Interestingly, polyurethanes containing P(3FOx-BrOx) have higher theta(adv) and lower theta(rec) than the homo-telechelic PUs [P(3FOx) = poly(2-methyl-2-trifluoroethoxymethyl-1,3 propylene oxide; P(BrOx) = poly(2-methyl-2-bromomethyl-1,3-propylene oxide)]. For IPDI-BD(40)/P(3FOx/BrOx-1:1), theta(adv) (116 degrees) is higher and theta(rec) (32 degrees) is lower (Deltatheta, 84 degrees ) than any other homo- or co telechelic polyurethane. The unusual wetting behavior for P(FOx/BrOx) polyurethanes is correlated with FOx-BrOx dyad content, and a reversible H bonding mechanism is proposed to explain the results. PMID- 16262348 TI - Biotin-streptavidin-induced aggregation of gold nanorods: tuning rod-rod orientation. AB - We report herein biotin-streptavidin-mediated aggregation studies of long gold nanorods. We have previously demonstrated end-to-end linkages of gold nanorods driven by the biotin-streptavidin interaction (Caswell et al. J. Am. Chem. Soc. 2003, 125, 13914). In that report, the specific binding of biotin disulfide to the gold nanorod edges was achieved due to the preferred binding of thiol molecules to the Au[111] surface (gold nanorod ends) as opposed to the gold nanorod side faces. This led to the end-end linkage of gold nanorods upon subsequent addition of streptavidin. In this report we demonstrate a simple procedure to biotinylate the entire gold nanorod surface and subsequently form a 3-D assembly by addition of streptavidin. Gold nanorods were synthesized by the three-step seeding protocol documented in our previous articles. The surface of gold nanorods was further modified by a layer of a weak polyelectrolyte, poly(acrylic acid), PAA. A biotin molecule which has an amine group at one end (biotin-PEO-amine) was anchored to the carboxylic acid group of the polyelectrolyte using the well-known carbodiimide chemistry. This process biotinylates the entire gold nanorod surface. Addition of streptavidin further leads to aggregation of gold nanorods. A closer look at the aggregates reveals a preferential side-to-side assembly of gold nanorods. The gold nanorods were characterized at each stage by UV-vis spectroscopy, light scattering, and transmission electron microscopy (TEM) measurements. PMID- 16262349 TI - Layered polyelectrolyte-silica coating for nanocapsules. AB - This paper demonstrates the ability to grow silica directly on a deposited surface of polyelectrolyte. Using this strategy, we describe the deposition of layered polyelectrolyte-silica coating on negatively charged surfaces of polystyrene particles and latex nanocapsules, which could not be coated directly with silica alone. By etching away the underlying polystyrene bead, we were able to form polyelectrolyte-silica capsules that were mechanically robust. Using scanning and transmission electron microscopy, we imaged and studied the coating after the deposition of each layer of polyelectrolyte and silica. We then applied this new coating to latex nanocapsules that were loaded with fluorescein molecules. We found that the coating procedure did not cause the loaded molecules to leak out from the capsules, and we determined that the variation in the number of loaded molecules among capsules arose from differences in the volume of the nanocavities and was not caused by the loading and coating of the capsules. This layered architecture permits the thickness of the coating to be controlled in principle over a wide dynamic range, but more importantly, this coating could act as an effective seal to prevent undesired leakage from nanocapsules and thus increase the long-term storability of loaded capsules. Over a 30-day period, we determined that leakage from uncoated capsules was significant but negligible for ones that were coated with two layers of polyelectrolyte-silica. Using single pulse UV photolysis of individual nanocapsules, we demonstrate that the molecules contained within coated capsules could be released effectively and on demand with a single laser pulse. PMID- 16262350 TI - Connected open structures from close-packed colloidal crystals by hyperthermal neutral beam etching. AB - We report the fabrication of connected open structures from close-packed colloidal crystals by hyperthermal neutral beam etching. Colloidal crystal films of polystyrene microspheres were prepared by a vertical deposition method. Exposure of the colloidal crystal films to hyperthermal neutral beam made isolated microspheres in the face-centered cubic lattice, each of which was connected with its twelve nearest neighbors through very thin cylinders. Due to the charge neutrality of impinging gas molecules of the hyperthermal neutral beam, the spherical shape of polymer microspheres was almost maintained during the etching process. The Bragg reflection peaks were modulated by the etched volume of colloidal crystals. Finally, the inverse structures of such open structures were replicated by a simple room-temperature chemical vapor deposition and subsequently burning out polymer template spheres. PMID- 16262351 TI - Low-temperature activation conditions for PAMAM dendrimer templated Pt nanoparticles. AB - Surface immobilized polyamidoamine (PAMAM) dendrimer templated Pt nanoparticles were employed as precursors to heterogeneous catalysts. CO oxidation catalysis and in situ infrared spectroscopy were used to evaluate conditions for dendrimer removal. Infrared spectroscopy showed that PAMAM dendrimer amide bonds begin decomposing at temperatures as low as 75 degrees C. Although the amide stretches are completely removed after 3 h of oxidation at 300 degrees C, 16 h were required to reach maximum catalytic activity. Further treatment under oxidizing or reducing atmospheres did not cause substantial changes in activity. Infrared spectroscopy of the activated materials indicated that organic residues, probably surface carboxylates, are formed during oxidation. These surface species passivate the Pt NPs, and their removal was required to fully activate the catalyst. Substantially less forcing activation conditions were possible by employing a CO/O(2)/He oxidation treatment. At appropriate temperatures, CO acts as a protecting group for the Pt surface, helping to prevent fouling of the nanoparticle by organic residues. CO oxidation catalysis and infrared spectroscopy of adsorbed CO indicated that the low temperature activation treatment yielded supported nanoparticles that were substantially similar to those prepared with more forcing conditions. PMID- 16262352 TI - Reversible aggregation of polystyrene-block-poly(2-vinylpyridine)-block poly(ethylene oxide) block copolymer micelles in acidic aqueous Solutions. AB - Micelles of polystyrene-block-poly(2-vinylpyridine)-block-poly(ethylene oxide) (PS-PVP-PEO) were studied in acidic aqueous solutions by static and dynamic light scattering, alkalimetric titration, fluorescence correlation spectroscopy, and after deposition on a mica surface by atomic force microscopy. The PS-PVP-PEO micelles prepared by dialysis in ternary 1,4-dioxane-methanol-acidic water mixtures have a very low association number and show a strong tendency to form aggregates. The aggregation, which is promoted at low pH, seems to be fully reversible. Possible mechanisms of the aggregation are discussed. Atomic force microscopy scans of PS-PVP-PEO micelles deposited on a mica surface reveal the formation of micellar aggregates and support the general concept of aggregation upon changes in conditions and deterioration of the stability of small micelles. PMID- 16262353 TI - Sub-micrometer-sized metal tubes from electrospun fiber templates. AB - Metallic tubes have been synthesized by a polymer-based template approach using electroless deposition. Gold, copper, and nickel were deposited as thin films on sub-micrometer polymer fibers which ranged in diameter from approximately 160 to 400 nm. After thermal degradation of the template fibers at 300 and 650 degrees C, tubes between 450 and 730 nm were obtained with wall thicknesses of 50-150 nm. Characterization by scanning electron microscopy, energy dispersive spectroscopy, transmission electron microscopy, Fourier transform infrared spectroscopy, and powder X-ray diffraction indicate that the tubes have a face-centered cubic structure with [111] preferred orientation for all of the metals investigated and that the tube walls are polycrystalline, composed of nanoparticles, ranging in size from 5.0 to 25.0 nm. PMID- 16262354 TI - Poly(thieno[3,4-b]thiophene)-poly(styrene sulfonate): a low band gap, water dispersible conjugated polymer. AB - Herein we report the oxidative chemical polymerization of thieno[3,4-b]thiophene (T34bT) using several different oxidants including ferric sulfate, ammonium persulfate, and hydrogen peroxide in the presence of poly(styrenesulfonic acid) in water and properties of the resulting poly(thieno[3,4-b]thiophene) poly(styrenesulfonic acid) (PT34bT-PSS) dispersion. The PT34bT-PSS is rendered a colloidal dispersion in water with a particle size diameter ranging between 180 and 220 nm depending on the oxidant used for polymerization. PT34bT-PSS films have band gaps of ca. 1 eV (1260 nm) as determined by the onset of the pi to pi transition from the vis-NIR spectrum with absorption maxima ranging from 1.4 eV (912 nm) to 1.7 eV (724 nm). The neutral and oxidized forms of PT34bT-PSS prepared from ferric sulfate dispersed in water were blue and lime green, respectively, whereas the neutral and oxidized forms of PT34bT-PSS prepared from ammonium persulfate and hydrogen peroxide were blue and blue-green, respectively. Spectral properties of the PT34bT-PSS dispersion can be tuned by the combination of oxidants. PT34bT-PSS films showed ca. 100% cation dominant ion transport behavior as determined by electrochemical gravimetry with each charge-discharge cycle and the doping level of the polymer was calculated to be 26%. Electrical conductivities for these polymers were found to be dependent on chemical oxidants used and varied from 10(-2) to 10(-4) S/cm. PMID- 16262355 TI - Stepped light-induced transient measurements of photocurrent and voltage in dye sensitized solar cells: application for highly viscous electrolyte systems. AB - To measure electron diffusion coefficients (D) and electron lifetimes (tau) of dye-sensitized solar cells (DSC), we introduced stepped light-induced transient measurements of photocurrent and voltage (SLIM-PCV), which can simplify the optical setup and reduce measurement time in comparison to conventional time-of flight and frequency-modulated measurements. The method was applied to investigate the influence of the viscosity of a thermally stable high-boiling point solvent on the energy conversion efficiency of DSCs. By systematic study of the influence of the viscosity, the species of cations as the counter charge of I(-)/I(3)(-), and the concentrations of electrolytes, we concluded that a lower dye cation reduction rate due to slower iodine diffusion is a limiting factor for a highly viscous electrolyte system. On the other hand, comparable values of D and increased values of tau were observed in a highly viscous electrolyte. By employing 0.5 M TBAI and 0.05 M I(2) in propylene carbonate, the efficiency of the DSC became comparable to that of a DSC using conventional electrolytes consisting of LiI, imidazolium iodide, and 4-tert-butylpyridine in methoxyacetonitrile. The simultaneous evaluation of D and tau through the appropriately simple measurement realizes fast optimization of the efficient and reliable DSC composed of thermally stable but often viscous electrolytes. PMID- 16262356 TI - Biopolymer and carbon nanotubes interface prepared by self-assembly for studying the electrochemistry of microperoxidase-11. AB - Stable films of biopolymer chitosan and carbon nanotubes were prepared by a layer by-layer self-assembly technique. Atomic force microscopy, scanning electron microscopy, cyclic voltammetry, and UV-vis spectroscopy were used to characterize the film assembly. Atomic force microscopy and scanning electron microscopy showed that an even, stable film was formed. The UV-vis spectroscopy and cyclic voltammetry study indicated the uniform growth of the film. The property of the self-assembled multilayer film in promoting protein electron transfer was demonstrated by incorporating microperoxidase-11 in the film. Microperoxidase-11 in the multilayer film could transfer electrons with the electrode indicating that carbon nanotubes could wire the protein to the electrode. The electrocatalytic activity of the microperoxidase-11 containing multilayer film modified electrode toward H(2)O(2) and O(2) was investigated. The results showed that along with the increase in the assembled layers the electrocatalytic reduction potentials of H(2)O(2) and O(2) shifted positively. The prepared multilayer film of chitosan and carbon nanotubes containing protein was a sensitive interface for electrocatalytic study. PMID- 16262357 TI - Directed growth of pure phosphatidylcholine nanotubes in microfluidic channels. AB - The morphology of self-assembled phospholipid membranes (e.g., micelles, vesicles, rods, tubes, etc.) depends on the method of formation, secondary manipulation, temperature, and storage conditions. In this contribution, microfluidic systems are used to create pure phosphatidylcholine (PC) micro- and nanotubes with unprecedented lengths. Tubes up to several centimeters in length and aligned with the long axis of the microchannel were created from spots of dry films of 1,2-dilauroyl-sn-glycero-3-phosphocholine (DLPC), 1,2-dimyristoyl-sn glycero-3-phosphocholine (DMPC), and 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC). These high aspect ratio structures, which, to our knowledge, represent the first examples of extended tubes formed from pure PC lipids, were examined by fluorescence microscopy, electron and optical microscopy, and optical manipulation tools (i.e., a laser trap and laser scalpel) to characterize structure and stability. In particular, the tubular structure was confirmed by observation of fluorescent dyes that were sequestered within the aqueous cavity or within the phospholipid tube. Compared to other phospholipid tubes, the tubes formed from PC lipids in microfluidic channels show high mechanical stability and rigidity that depend on tube size, age, and storage conditions. PMID- 16262358 TI - Rapid polymer brush growth by TEMPO-mediated controlled free-radical polymerization from swollen plasma deposited poly(maleic anhydride) initiator surfaces. AB - Pulsed plasma-chemical deposition of poly(maleic anhydride) is shown to be a substrate-independent method for functionalizing solid surfaces with initiator sites for nitroxide-mediated controlled free-radical graft polymerization. Swelling of the initiator film via aminolysis can lead to grafted polymer brushes that are 1 order of magnitude thicker than those obtained by existing methods on solid surfaces. PMID- 16262359 TI - Organo-Laponites as novel mesoporous supports for manganese(III) salen catalysts. AB - A Mn(III) salen complex was immobilized onto the Laponite surface using three different methodologies: method A, direct immobilization of the complex on the parent Laponite; method B, covalent anchoring through cyanuric chloride (CC); and method C, covalent anchoring through CC into a 3-aminopropyl)triethoxysilane (APTES) modified Laponite. All of the materials were characterized by FTIR, XPS, thermogravimetry, XRD, and nitrogen isotherms at 77 K, to gather information on the modifications introduced by the organo spacers within the Laponite surface, as well as on the anchored complex integrity; the Mn based materials were screened in the heterogeneous epoxidation of styrene. The results have shown that the immobilization of the manganese(III) salen complex by methods B and C have occurred at the edges of the clay particles through the spacers (APTES and CC) that have been anchored onto the Si-OH groups, whereas in method A, the complex is distributed throughout the clay surface, including the interlayer region. Therefore, the manganese loadings on the Laponites were as follows: materials prepared by method A >> method B > method C. All of the heterogeneous catalysts showed high styrene epoxide selectivity, with that prepared by method A showing comparable styrene epoxide selectivity as the homogeneous phase reaction. The styrene epoxide yields decrease in the following order: materials prepared by method A > method B > method C (1st cycles), which parallel the respective support catalytic activity and decreasing of manganese content. The heterogeneous catalysts prepared using methods B and C could be reused at least for four times, with the former exhibiting the most stable catalytic activity, but that prepared by method A showed a significant decrease after two catalytic cycles. PMID- 16262360 TI - Small-angle neutron scattering on a core-shell colloidal system: a contrast variation study. AB - Small-angle neutron scattering (SANS) measurements are reported on a sterically stabilized, core-shell colloidal system using contrast variation. Aqueous dispersions of polystyrene particles bearing grafted poly(ethylene glycol) (PEG) have been studied over a large range of particle concentrations and two different solvent conditions for the PEG polymer. SANS data are analyzed quantitatively by modeling the particles as core-shell colloids. In a good solvent and under particle contrast conditions, an effective hard-sphere interaction captures excluded-volume interactions up to high concentrations. Contrast variation, through isotopic substitution of both the core and solvent, expedite a detailed study of the PEG layer, both in the dilute limit and as a function of the particle concentration. Upon diminishing the solvent quality, subtle changes in the PEG layer translate into attractions among particles of moderate magnitude. PMID- 16262361 TI - Carbon microspheres with supported silver nanoparticles prepared from pollen grains. AB - In this paper, we present a new method to fabricate carbon microspheres with supported silver nanoparticles on the surfaces. In this method, pollen grains were first treated with AgNO(3) aqueous solution, then preoxidized in air at 300 degrees C and carbonized in nitrogen at 600 degrees C, resulting in the silver/carbon nanocomposites. The silver/carbon nanocomposites were characterized by means of SEM, TEM, TG, and XRD. The size and distribution of the silver nanoparticles on the carbon microsphere surface could be controlled by tuning the AgNO(3) treatment conditions. PMID- 16262362 TI - Molybdenum carbide formation in molybdenum-doped organic and carbon aerogels. AB - A Mo-doped organic aerogel and its corresponding carbonized derivative at 1000 degrees C were obtained. Both samples were treated in a H(2)/Ar flow. After this treatment, a mixture of Mo(VI) and Mo(2)C was detected in both samples. The results obtained indicate that the presence of H(2) in the gas flow is necessary to obtain the carbide phase, due to the formation of CH(4) or even CH(x)() species that reduce and carburize the molybdenum oxide phase. Carburization of the Mo-doped organic aerogel yielded better results compared with carburization of the Mo-doped carbon aerogel. PMID- 16262363 TI - Computational predictions of stable 2D arrays of bidisperse particles. AB - We study computationally the stability of various 2D arrays of bidisperse mixtures of stabilized nanoparticles through a melting simulation employing the Metropolis algorithm for determining surface diffusion. In our previous work [Langmuir 2004, 20, 9408], we studied computationally the stability of bidispersed monolayers of thiol-stabilized gold nanoparticles with a size ratio (sigma) of 0.375. We found that interparticle forces were essential to stabilize the LS (the two-dimensional NaCl analogue) lattice at the experimentally determined surface coverage. In this paper, we extend our study to determine the conditions necessary to form stable LS(2), LS(4), and LS(6) lattices, which have yet to be observed. Using a simple design rule that involves matching the distances between either large-large particles and large-small particles or large small particles and small-small particles to correspond to the respective potential minima leads to predictions for size ratios that will form each desired lattice, given other parameters characterizing the systems' physical properties. We predict and verify computationally LS(2), LS(4), and LS(6) lattices at relatively low surface coverages. Additional simulations show that the LS, LS(2), and LS(6) lattices are indeed stable structures at their predicted surface coverage, whereas the LS(4) lattice is a metastable structure; however, a modest increase in the surface coverage of the LS(4) lattice converts it to a stable rather than long-lived metastable structure. This study may be used as a guide for experimentalists in their search for these novel structures. PMID- 16262364 TI - Influence of added particles on the phase behavior of polymer solutions. Analysis by mean-field lattice theory. AB - The influence of added colloidal particles on the phase stability of polymer solutions is investigated theoretically. The polymer has an affinity to the particle surface. A mean-field lattice theory based on the Flory-Huggins theory is used to calculate the phase behavior in solutions containing a single polymer component and particles. The particles are described in two different ways. The first approach considers the surface free energy associated with added solid particles and the mixing entropy of the particles. In the second approach, the particles are simply modeled as large polymers. Both ways of describing the added particles show that the added particles decrease the stability of the polymer solution when the polymer-particle attraction is strong. A higher particle concentration enhances the effect. Experiments where polystyrene latex particles are added at different concentrations to aqueous dispersions of ethyl(hydroxyethyl)cellulose (EHEC) support the theoretical findings. PMID- 16262365 TI - How many Langmuirs are required for monolayer formation? AB - In the present work, we provide the exact answer to the title question employing a probabilistic approach. The average number of Langmuirs L required for monolayer formation was found to be equal to (1/i), i.e., the armonic series up to the nth term, where n is the number of adsorption sites. This result is particularly useful when a reduced number of adsorption sites is considered, such as adsorption on small terraces of nanoscopic dimensions where the value of n could be in the range of a few thousands sites. In this case, the use of integrated equations derived from the mean-field approach would provide completely misleading results. PMID- 16262366 TI - Stability of Na-, K-, and Ca-montmorillonite at high temperatures and pressures: a Monte Carlo simulation. AB - Monte Carlo grand canonical molecular simulations on the hydration of Na-, K-, and Ca-montmorillonite show that between 333 and 533 K and 300-1300 bar Na montmorillonite forms stable one-layer hydrates of d(001) spacings 12.64-12.38 Angstroms, K-montmorillonite of 12.78-12.59 Angstroms, and Ca-montmorillonite of 12.48-12.32 Angstroms. A two-layer hydrate of 14.80 Angstroms occurs for Na montmorillonite at 533 K and 1300 bar, for K-montmorillonite of 15.32 Angstroms at 533 K and 1300 bar and of 14.74 Angstroms at 533 K and 2000 bar, and for Ca montmorillonite of 13.83 Angstroms at 473 K and 1000 bar. Three-layer hydrates may possibly form within these same ranges. Outside of them, one-layer hydrates simulate as the only stable hydrates. In sedimentary basins, the two-layer hydrate of Ca-montmorillonite will locate at 6.7 km depth and those of Na- and K montmorillonite at 8.7 km depth; above and below these depths, the one-layer hydrates are the stable phases. PMID- 16262367 TI - Two-dimensional supercritical behavior of an ethanol monolayer: a molecular dynamics study. AB - The two-dimensional (2D) supercritical behavior of an ethanol monolayer formed at the vapor/liquid interface of an ethanol solution has been investigated by a molecular dynamics (MD) calculations with a combination of the OPLS-UA and SPC/E potential models. A 100 A thick slab of ethanol solution was placed at the volume center of the rectangular unit cell by 10 A thick nonabsorbate water surfaces. With such an initial configuration, five independent 15 ns NVT constant MD calculations were carried out under 298.15 K, in which the initial ethanol mole fraction of the bulk solution layer was set to 0.010, 0.022, 0.045, 0.10, and 0.20, respectively. The 2D radial distribution function (rdf) of an adsorbed ethanol molecule showed that the ethanol monolayer could be regarded as a 2D fluid where the adsorbed ethanol molecule had an effective 2D diameter of 4.65 A. On the basis of the 2D rdf result, 2D cluster analysis was carried out from the perspective of the percolation theory. It is confirmed that the critical area occupation probability density, the critical exponents, and the fractal dimension of both nonpercolating and percolating clusters satisfied their nature of universality. Therefore, we concluded that an ethanol monolayer formed at the vapor/liquid interface of ethanol solution behaves as a 2D supercritical fluid at 298.15 K. PMID- 16262368 TI - Comment on breakdown of colloid filtration theory: role of the secondary energy minimum and surface charge heterogeneities. PMID- 16262370 TI - Comment on epitaxial relationships between uric acid crystals and mineral surfaces: a factor in urinary stone formation. PMID- 16262371 TI - Complementary matching in domain formation within lipid bilayers. AB - Domain structure and formation in lipid bilayers are investigated by molecular dynamics simulations using a coarse-grained lipid model. The lipid bilayers consist of two lipid types that are identical except for tail length. At a temperature intermediate to the two melting temperatures of the constituent lipid types, gel domains spontaneously form from an initial random structure. The simulations reveal that the gel domains consist of both lipid types in a complementary match. If a long lipid is in the top monolayer, then a short lipid is underneath and vice versa. The gel domains have a larger thickness than the surrounding liquid phase. The thickness of the gel domains is close to that of the pure long lipid gel phase bilayers. However, since in the mixed gel domains the lipids are not tilted and in the pure gel phase the lipids are tilted, the two thicknesses are similar, and the underlying structure is therefore not distinguishable solely by thickness measurements. PMID- 16262372 TI - Lacticin 481 synthetase phosphorylates its substrate during lantibiotic production. AB - Lacticin 481 synthetase (LctM) catalyzes the ATP-dependent conversion of a ribosomally synthesized peptide to a polycyclic thioether antibiotic. It is a bifunctional enzyme that dehydrates four Ser/Thr residues to the corresponding dehydro amino acids and catalyzes the conjugate addition of Cys residues to these dehydro residues in a regio- and stereoselective process. We show here that incubation of truncated substrates with LctM results in products that are phosphorylated in the region of dehydration. Furthermore, synthetic peptides containing phosphorylated Ser and/or Thr residues are accepted by the enzyme as substrates resulting in the elimination of phosphate and dehydro amino acid production. This activity is only observed if ADP is added as cosubstrate. These results argue strongly that the enzyme utilizes ATP to phosphorylate the Ser/Thr residues that are targeted for dehydration. ATP does not appear to be required for peptide translocation or cyclization. PMID- 16262373 TI - Water molecules in a protein cavity detected by a statistical-mechanical theory. AB - Four water molecules confined in a small cavity of hen egg-white lysozyme were detected by means of the three-dimensional (3D) RISM theory, a statistical mechanical theory of molecular solutions. This is the first theoretical realization of confined molecules in a protein without making nonsense tricks, such as placing the molecules in the space a priori. Possible impacts which the result may have on biochemistry and biophysics, including the molecular recognition, enzymatic reactions, etc., are discussed. PMID- 16262374 TI - Quasi-continuous growth of ultralong carbon nanotube arrays. AB - We report the growth of ultralong (>10 cm) multi-walled and single-walled carbon nanotubes such that the length is limited by the size of the furnace rather than by the termination of growth. The disturbance of microscale laminar flows results in disordered or shorter growth of carbon nanotubes. By downsizing reaction pipes, reaction gas flows are stabilized with low Reynolds numbers. In this way, the catalyst nanoparticles at the end of growing carbon nanotubes can travel a longer distance to grow ultralong nanotubes. PMID- 16262375 TI - Stable, reproducible nanorecording on rotaxane thin films. AB - Stable, reversible conductance transitions and memory effects were observed in solid-state rotaxane-based Langmuir-Blodgett (LB) thin films. Taking advantage of the switching property, we have achieved reproducible nanometer-scale recording dots on rotaxane LB films via voltage application using a scanning tunneling microscope probe. The stable and easily controlled conductance switching makes the rotaxanes a hopeful candidate for nanorecording. PMID- 16262376 TI - Bromine is an endogenous component of a vanadium bromoperoxidase. AB - On the basis of EXAFS and MS/MS experimental results and a reinterpretation of the electron density map obtained by X-ray crystallography, we describe a new post-translational modification, that is, a 3,5-dibromotyrosine residue that is incorporated in the polypeptide chain of a vanadium haloperoxidase. PMID- 16262377 TI - Sensitization of nanocrystalline TiO2 with black absorbers based on Os and Ru polypyridine complexes. AB - New metal complexes of the type [M(H3tcterpy)LY]+ (where M = Os(II) or Ru(II), L = substituted or unsubstituted bipyridine or pyridylquinoline, and Y = Cl-, I-, or SCN-) have been designed, synthesized, and characterized in view of their application for dye-sensitized solar cells (DSSCs). The Os dyes show a very broad absorption, and correspondingly, the DSSCs show an unprecedented spectral response in the NIR, with an onset at 1100 nm, reaching values of about 50% at 900 nm. The integrated photocurrent of some of such Os dyes is similar to that of the well-known [Ru(Htcterpy)(NCS)3](TBA) and superior to that of the [Ru(Hdcbpy)2(NCS)2](TBA)2 sensitizer. The Ru dyes show absorption and DSSC spectral response between those of the red and black dyes. Their advantage is their potential superior stability due to the reversible oxidative electrochemistry. PMID- 16262379 TI - The Tolman length: is it positive or negative? AB - By means of a large-scale molecular dynamics simulation, we show that the Tolman length, although positive, is much smaller in magnitude than previously reported. We found that the range of interparticle interaction can significantly affect the magnitude of the Tolman length. When the range of interaction is longer than five molecular diameters, the Tolman length is on the order of a few hundredths of the molecular diameter, rather than a few tenths known previously. PMID- 16262378 TI - A proposal for the mechanism-of-action of diazoparaquinone natural products. AB - Treatment of the representative diazoparaquinone prekinamycin dimethyl ether with Bu3SnH/AIBN in aromatic solvents furnishes moderate-to-good yields of formal aryl adducts wherein a molecule of solvent is attached to the carbon (C(11)) previously bearing the diazo function. Substituent studies provide evidence in support of a radical aromatic substitution mechanism, in which radical addition to the diazoparaquinone function generates an intermediate C(11) vinylic radical. PMID- 16262380 TI - Direct electrodeposition of highly dense Bi/Sb superlattice nanowire arrays. AB - Ordered arrays of Bi/Sb superlattice nanowires with diameters of about 50 nm have been produced by pulsed electrodeposition technique into the pores of anodic alumina membrane (AAM). The structure of Bi/Sb superlattice nanowire can be modulated by controlling the electrodeposition conditions. PMID- 16262382 TI - Synthesis, resolution, and aldol reactions of a planar-chiral Lewis acid complex. AB - The synthesis and resolution of a new planar-chiral Lewis acid complex is described. The compound is applied to asymmetric Mukaiyama aldol reactions, leading to the formation of the desired product with good stereoselectivity. The sense of stereoselection is as predicted by the design, which exploits the ability of the Lewis acid to simultaneously serve as a sigma and a pi acceptor. Mechanistic observations are consistent with rate-determining formation of an aldehyde-Lewis acid complex, followed by rapid nucleophilic addition. PMID- 16262381 TI - Functional GPCR microarrays. AB - This paper describes G-protein-coupled receptor (GPCR) microarrays on porous glass substrates and functional assays based on the binding of a europium-labeled GTP analogue. The porous glass slides were made by casting a glass frit on impermeable glass slides and then coating with gamma-aminopropyl silane (GAPS). The emitted fluorescence was captured on an imager with a time-gated intensified CCD detector. Microarrays of the neurotensin receptor 1, the cholinergic receptor muscarinic 2, the opioid receptor mu, and the cannabinoid receptor 1 were fabricated by pin printing. The selective agonism of each of the receptors was observed. The screening of potential antagonists was demonstrated using a cocktail of agonists. The amount of activation observed was sufficient to permit determinations of EC50 and IC50. Such microarrays could potentially streamline drug discovery by helping integrate primary screening with selectivity and safety screening without compromising the essential functional information obtainable from cellular assays. PMID- 16262383 TI - Regio- and stereoselective homocoupling of gamma-arylated tert-propargyl alcohols with liberation of a ketone molecule and successive cyclization to produce fluorescent dihydrofuran derivatives. AB - 1,1-Disubstituted 3-aryl-2-propyn-1-ols undergo unprecedented regio- and stereoselective homocoupling with liberation of a ketone molecule in the presence of a rhodium catalyst to give the corresponding 2-hydroxymethyl-(E)-enynes. The subsequent cyclization of the enynes in the presence of a base affords fluorescent 2,3-dihydrofuran derivatives. PMID- 16262384 TI - Blue ferrocenium azurin: an organometalloprotein with tunable redox properties. AB - A ferrocene derivative (2-[(methylsulfonyl)thio]ethylferrocene) (1) has been synthesized and incorporated into apo-azurin from Pseudomonas aeruginosa by covalent attachment to the highly conserved Cys112. The resulting artificial organometalloprotein (a protein containing organometallic compounds in the active site) has been characterized by UV-vis, electrospray mass spectrometry, and cyclic voltammetry (CV). Incorporation of 1 into azurin resulted in a higher solubility of the ferrocene group and improved stability of the ferrocenium species in aqueous solution, as shown by a more intense UV-vis absorption and a more reversible CV of the attached ferrocene group, respectively. The incorporation of 1 also increased the reduction potential of the complex from 402 to 579 mV (vs NHE), consistent with the ferrocene group being encapsulated inside the hydrophobic environment of the protein. Modulation of the reduction potential of ferrocene by residues near the secondary coordination sphere has also been demonstrated. Raising the pH from 4 to 9 resulted in a greater than 80 mV decrease in reduction potential of the protein-bound ferrocene (from 579 to 495 mV), while replacing Met121, an amino acid residue in close proximity to the ferrocene group with a positively charged Arg or negatively charged Glu, resulted in the predicted increase or decrease in reduction potential at all pH values. Similarly, substitution of Met121 with a more hydrophobic Leu raised the reduction potential. The increased solubility, stability, and tune-ability of this organometalloprotein make it an ideal choice for carrying out a number of biological reactions, such as long-range electron transfer or sensing. As an example of such applications, stoichiometric oxidation of ferrocytochrome c by the blue ferrocenium azurin was demonstrated. PMID- 16262385 TI - Polymer nano- and microspheres with bumpy and chain-segregated surfaces. AB - Reported is a novel and simple method for the preparation of polymer spheres bearing hemispherical surface bumps where one type of polymer chains concentrates. The method is used to produce spheres with a diameter between approximately 30 and approximately 500 nm. Spheres with chain-segregated bumpy surfaces may find applications in drug delivery and other areas. PMID- 16262386 TI - Exogenous nitrile substrate hydroxylation by a new dicopper-hydroperoxide complex. AB - A dicopper(I)/phenol-ligand complex in RCN solvents reacts with O2 producing a mu 1,1-hydroperoxo dicopper(II) species. Subsequent thermal transformation results in nitrile hydroxylation and elimination of cyanide, as revealed by the isolation in comparable yields of (i) a cyanide-bridged tetranuclear cluster complex and (ii) benzaldehyde (for R = PhCH2); 18O labeling confirms that the PhC(O)H oxygen atom derives from O2. PMID- 16262387 TI - Three-layer composite magnetic nanoparticle probes for DNA. AB - A method for synthesizing composite nanoparticles with a gold shell, an Fe3O4 inner shell, and a silica core has been developed. The approach utilizes positively charged amino-modified SiO2 particles as templates for the assembly of negatively charged 15 nm superparamagnetic water-soluble Fe3O4 nanoparticles. The SiO2-Fe3O4 particles electrostatically attract 1-3 nm Au nanoparticle seeds that act in a subsequent step as nucleation sites for the formation of a continuous gold shell around the SiO2-Fe3O4 particles upon HAuCl4 reduction. The three-layer magnetic nanoparticles, when functionalized with oligonucleotides, exhibit the surface chemistry, optical properties, and cooperative DNA binding properties of gold nanoparticle probes, but the magnetic properties of the Fe3O4 inner shell. PMID- 16262388 TI - Facile Ru-H2 heterolytic activation and intramolecular proton transfer assisted by basic N-centers in the ligands. AB - The use of the phosphine PPh2py instead of PPh3 in complexes of the type [Cp*RuH(P)2] enormously alters the kinetic control of the proton-transfer reactions over this compound and its chemical behavior. The reaction at low temperature of [Cp*RuH(PPh2py)2], 2, with HBF4 gives as products the classical dihydride trans-[Cp*RuH2(PPh2py)2](BF4), 3 (1 equiv of HBF4) or the dihydrogen bonded complex [Cp*RuHH(PPh2pyH)(PPh2py)](BF4)2, 4 (2 equiv of HBF4). These complexes exhibit very accessible intramolecular processes of proton transfer, and finally, a slow release of H2 takes place at room temperature. Derivatives 2 and 3 are active catalysts for the deuterium labeling of H2 using methanol-d4 as an isotopic source. This demonstrates that the release of hydrogen is reversible, that the heterolytic activation of H2 is an easy process, and that acid species participate in the intramolecular proton-transfer processes. These observations are supported by reaction-coordinate calculations at the DFT/B3LYP level that show the existence of a low-energy reaction path that easily transforms the classical trans dihydride complex into the nonclassical cis dihydrogen compound in a reversible way, through the involvement of hydrogen- and dihydrogen-bonded intermediates and the essential participation of the pyridine centers. The different energy minima of this reaction profile are very accessible through low energy transition states, all of which have been located. PMID- 16262390 TI - DNA single strands tethered to fused quartz/water interfaces studied by second harmonic generation. AB - Second harmonic generation (SHG) is used to study oligonucleotides at aqueous/solid interfaces for the first time. Detailed thermodynamic state information for interfacial DNA single strands, namely, the interfacial charge density, the interfacial potential, and the change in the interfacial energy density, is obtained. The phosphate groups on the DNA backbone serve as intrinsic labels that do not require DNA modification other than surface attachment. This approach is broadly applicable for the investigation of DNA during its interaction with biological targets, as well as charged biopolymers in general, and has important implications for predicting and controlling macromolecular interactions, improving biodiagnostics, and understanding life processes. PMID- 16262389 TI - E-olefin dipeptide isostere incorporation into a polypeptide backbone enables hydrogen bond perturbation: probing the requirements for Alzheimer's amyloidogenesis. AB - Herein, we report a stereospecific E-olefin dipeptide isostere synthesis that can be used to make gram quantities of the Phe-Phe isostere desired for eliminating a specific backbone H-bond donor and acceptor in the Alzheimer's disease related Abeta peptide. The Phe19-Phe20 E-olefin analogue of Abeta(1-40) was prepared by solid-phase peptide synthesis and was subjected to amyloidogenesis conditions. This analogue can aggregate into spherical morphologies but does not progress on to form protofibrils or fibrils as is the case for the all-amide sequence, providing insight into the structural requirements for amyloidogenesis. PMID- 16262391 TI - Nature of the cytochrome c molten globule. AB - We have employed fluorescence energy transfer (FET) kinetics to probe unfolded and molten globule states of five dansyl (Dns) variants of Saccharomyces cerevisiae iso-1 cytochrome c. The covalently bound Fe(III) heme group quenches Dns fluorescence by energy transfer; measurements of FET kinetics yielded distributions of D-A distances (P(r)) for these states. The P(r) distributions and corresponding mean force potentials (U(r)) show that the cytochrome c molten globule is a highly structured state with a substantial number of native interactions. Wide P(r) distributions directly reflect the dynamic nature and conformational diversity of this molten globule. P(r) distributions for the "burst-phase" refolding intermediate suggest that the equilibrium cytochrome c molten globule is not a suitable model for early intermediates formed during protein refolding. PMID- 16262392 TI - Polyether-bridged sexithiophene as a complexation-gated molecular wire for intramolecular photoinduced electron transfer. AB - The porphyrin-sexithiophene-fullerene triad 2, where the two central thiophene units of the sexithiophene spacer are bridged with a crown-ether-like polyether chain, undergoes efficient intramolecular electron transfer from the photoexcited porphyrin moiety to the fullerene through the sexithiophene. However, complexation with a sodium cation in the crown ether ring causes complete suppression of electron transfer as a result of a drastic conformational change of the sexithiophene backbone. Furthermore, decomplexation resumes the photoinduced electron transfer. This on/off switching phenomenon indicates that the polyether-bridged sexithiophene can function as a complexation-gated molecular wire. PMID- 16262393 TI - Mechanism for reduction catalysis by metal oxo: hydrosilation of organic carbonyl groups catalyzed by a rhenium(V) oxo complex. AB - The rhenium oxo complex [Re(O)(hoz)2][TFPB], 1 (where hoz = 2-(2'-hydroxyphenyl) 2-oxazoline(-) and TFPB = tetrakis(pentafluorophenyl)borate) catalyzes the hydrosilation of aldehydes and ketones under ambient temperature and atmosphere. The major organic product is the protected alcohol as silyl ether. Isolated yields range from 86 to 57%. The reaction requires low catalyst loading (0.1 mol %) and proceeds smoothly in CH2Cl2 as well as neat without solvent. In the latter condition, the catalyst precipitates at the end of reaction, allowing easy separation and catalyst recycling. Re(O)(hoz)(H), 3, was prepared, and its involvement in an ionic hydrosilation mechanism was evaluated. Complex 3 was found to be less hydridic than Et3SiH, refuting its participation in catalysis. A viable mechanism that is consistent with experimental findings, rate measurements, and kinetic isotope effects (Et3SiH/Et3SiD = 1.3 and benzaldehyde H/benzaldehyde-D = 1.0) is proposed. Organosilane is activated via eta2 coordination to rhenium, and the organic carbonyl adds across the coordinated Si H bond [2 + 2] to afford the organic reduction product. PMID- 16262394 TI - A stable fused bicyclic disilene as a model for silicon surface. AB - We synthesized the first fused bicyclic disilene 1 representing topologically a partial structure of the Si(001) surface up to the third layer. In the solid state, the five-membered rings adopt the envelope conformation, and the Si=Si double bond in 1 exists in the slightly cis-bent form (bent angle theta is 3.6 degrees ) compared to that of the highly cis-bent dimer on the Si(001) surface. Highly symmetric 1H NMR spectral pattern of 1 remains even at -80 degrees C, indicating the facile ring flipping of the bicyclic skeleton in solution. While syn-adduct was obtained in the reaction of 1 with water, anti-addition of chlorine atoms across the Si=Si double bond in 1 was observed in the reaction with carbon tetrachloride. The structural characteristics of the 9,10 phenanthrenequinone adduct 7 are in good accord with those of the proposed structure of the 9,10-phenanthrenequinone molecule adsorbed on the Si(001) surface. PMID- 16262395 TI - White-light emission from magic-sized cadmium selenide nanocrystals. AB - Magic-sized cadmium selenide (CdSe) nanocrystals have been pyrolytically synthesized. These ultra-small nanocrystals exhibit broadband emission (420-710 nm) that covers most of the visible spectrum while not suffering from self absorption. This behavior is a direct result of the extremely narrow size distribution and unusually large Stokes shift (40-50 nm). The intrinsic properties of these ultra-small nanocrystals make them an ideal material for applications in solid state lighting and also the perfect platform to study the molecule-to-nanocrystal transition. PMID- 16262396 TI - Initial example of a triangular single-molecule magnet from ligand-induced structural distortion of a [MnIII3O]7+ complex. AB - The reaction of [Mn3O(O2CR)6(py)3](ClO4) (R = Me, Et) with methyl 2-pyridyl ketone oxime (mpkoH) in a 1:3 molar ratio in MeOH/MeCN leads to [Mn3O(O2CR)3(mpko)3](ClO4) in 80-90% isolated yield. Ferromagnetic exchange interactions between the three MnIII ions in the nonplanar [MnIII3O]7+ triangular core lead to a spin ground state of S = 6; single-crystal studies reveal the temperature and sweep rate dependent hysteresis loops expected for a single molecule magnet. PMID- 16262397 TI - Catalytic Ni/Cr-mediated macrocyclization without use of high-dilution techniques. AB - The feasibility of catalytic Ni/Cr-mediated macrocyclization was demonstrated for the two substrates chosen from the halichondrin area. With 5 mol % of Ni and Cr catalysts, the macrocyclization was realized without use of high-dilution techniques. The reported method has a number of appealing features, including user-friendliness, easy workup, apparent scalability, and cost-effectiveness. In addition, all the required reagents are commercially available or obtainable in one or two steps from commercially available chemicals. PMID- 16262398 TI - Modeling side-on NO coordination to type 2 copper in nitrite reductase: structures, energetics, and bonding. AB - DFT calculations reveal the existence of metastable side-on {CuNO}10 and {CuNO}11 species relevant to the type 2 copper site of nitrite reductase (CuNIR). Side-on NO coordination seems especially favorable in energy terms for the {CuNO}11 species. The {CuNO}11 geometry parameters also seem to be in better agreement with those reported for a crystallographically characterized CuNIR intermediate, relative to the {CuNO}10 parameters. PMID- 16262399 TI - Proteasome inhibition by a totally synthetic beta-lactam related to salinosporamide A and omuralide. AB - A new and effective proteasome inhibitor, beta-lactam 3, has been accessed enantioselectively by multistep synthesis from the readily prepared intermediates 7 and 8 which were joined by a [2 + 2]-cycloaddition reaction to form the spiro beta-lactam 9 stereoselectively. The intermediate 9 was converted to 3 in seven steps and 30% overall yield. The beta-lactam 3 is stable for many days in water at pH 7, in contrast to the natural beta-lactones salinosporamide A (1) and omuralide (2). In common with 1 and 2, the beta-lactam 3 effectively inhibits the mammalian proteasome. PMID- 16262400 TI - Broad spectral domain fluorescence wavelength modulation of visible and near infrared emissive polymersomes. AB - Incorporation of an extended family of multi[(porphinato)zinc(II)] (PZn)-based supermolecular fluorophores into the lamellar membranes of polymersomes (50 nm to 50 mum diameter polymer vesicles) gives rise to electrooptically diverse nano-to micron (meso) scale soft materials. Studies that examine homogeneous suspensions of 100 nm diameter emissive polymersomes demonstrate fluorescence energy modulation over a broad spectral domain of the visible and near-infrared (600-900 nm). These polymersomal structures highlight that the nature of intermembranous polymer-to-fluorophore contacts depends on the position and identity of the porphyrins' phenyl ring substituents. Emissive polymersomes are shown to possess reduced spectral heterogeneity with respect to the established optical signatures of these PZn-based supermolecular fluorophores in solution; additionally, selection of fluorophore ancillary substituents predictably controls the nature of polymer-emitter noncovalent interactions to provide an important additional mechanism to further modulate the fluorescence band maxima of these meso-scale emissive vesicles. PMID- 16262401 TI - Oxaziridine-mediated catalytic hydroxylation of unactivated 3 degrees C-H bonds using hydrogen peroxide. AB - The design, structural characterization, and evaluation of a unique class of 1,2,3-benzoxathiazine-based oxaziridines as potent O-atom transfer agents for catalytic C-H hydroxylation and alkene epoxidation are described. Turnover of this reaction is made possible by employing a diaryl diselenide cocatalyst and urea.H2O2 as the terminal oxidant. Oxidation of saturated hydrocarbons is strongly biased toward 3 degrees C-H bonds even in systems possessing a significantly greater number of methylene groups. In addition, the benzoxathiazine catalyst is effective for epoxidation of terminal and electron deficient olefins. Collectively, these findings represent an important first step toward the advancement of general methodology for selective C-H oxidation. PMID- 16262402 TI - Enantioselective total syntheses of welwitindolinone A and fischerindoles I and G. AB - The first total syntheses of welwitindolinone A and the most complex members of the fischerindole family, fischerindoles I and G, are reported. Highlights of these short, protecting-group-free syntheses include the application of the recently developed direct indole-carbonyl coupling, a simple approach for installing the quaternary center with neighboring chlorine atom, a regioselective dehydrogenation/dehydration cascade to access fischerindole I, and a remarkably facile oxidative ring contraction to construct welwitindolinone A. An alternative biogenetic hypothesis, whose accuracy is suggested by the success of the current syntheses, is also put forth for this alkaloid family. PMID- 16262403 TI - Ab initio QM/MM study of class A beta-lactamase acylation: dual participation of Glu166 and Lys73 in a concerted base promotion of Ser70. AB - Beta-lactamase acquisition is the most prevalent basis for Gram-negative bacteria resistance to the beta-lactam antibiotics. The mechanism used by the most common class A Gram-negative beta-lactamases is serine acylation followed by hydrolytic deacylation, destroying the beta-lactam. The ab initio quantum mechanical/molecular mechanical (QM/MM) calculations, augmented by extensive molecular dynamics simulations reported herein, describe the serine acylation mechanism for the class A TEM-1 beta-lactamase with penicillanic acid as substrate. Potential energy surfaces (based on approximately 350 MP2/6-31+G calculations) reveal the proton movements that govern Ser70 tetrahedral formation and then collapse to the acyl-enzyme. A remarkable duality of mechanism for tetrahedral formation is implicated. Following substrate binding, the pathway initiates by a low energy barrier (5 kcal mol(-1)) and an energetically favorable transfer of a proton from Lys73 to Glu166, through the catalytic water molecule and Ser70. This gives unprotonated Lys73 and protonated Glu166. Tetrahedral formation ensues in a concerted general base process, with Lys73 promoting Ser70 addition to the beta-lactam carbonyl. Moreover, the three-dimensional potential energy surface also shows that the previously proposed pathway, involving Glu166 as the general base promoting Ser70 through a conserved water molecule, exists in competition with the Lys73 process. The existence of two routes to the tetrahedral species is fully consistent with experimental data for mutant variants of the TEM beta-lactamase. PMID- 16262404 TI - A strand-loop-strand structure is a possible intermediate in fibril elongation: long time simulations of amyloid-beta peptide (10-35). AB - A total of 6.2 micros molecular dynamics simulations of amyloid-beta (10-35) (Abeta) were performed in explicit water solvent. The results reveal that the collapsed-coil (cc) structure determined by experiments is stable at pH 5.6 for hundreds of nanoseconds, but it can exchange with a strand-loop-strand (SLS) structure on the microsecond time scale. The SLS structure has D23-K28 as a reverse loop and the central hydrophobic core and the C-terminal in hydrophobic contact. This SLS structure topologically resembles the proposed monomer conformation in fibrils. Since it has been suggested that a special conformation of Abeta is needed when the monomer binds to fibril ends to elongate fibrils, we propose that the SLS structure may be an important intermediate binding structure for Abeta fibril growth. Simulations at pH 2.0, which is used to mimic the mutation of E22Q and D23N, and at high temperature (400 K) indicate that the SLS structure is considerably populated under these conditions while the cc structure is disrupted. These results imply that the SLS structures may also be a binding intermediate in other conditions such as E22Q and/or D23N mutations and high temperature, which have been proved to promote fibril formation previously. PMID- 16262405 TI - Long-lived spin-correlated pairs generated by photolysis of naphthalene occluded in non-Bronsted acidic ZSM-5 zeolites. AB - Long-lived spin-correlated pairs were generated by laser irradiation of naphthalene (NAP) occluded as intact molecule within non-Bronsted acidic MnZSM-5 zeolites with MnSiO(2))(96-n)(AlO(2)n formula per unit cell. The laser UV photoionization generates primary NAP.+-electron pair as a fast phenomenon. These charge carriers exhibit lifetimes that extend over less than 1 h at room temperature and disappear according to two parallel competitive ways: direct charge recombination and electron transfer. This subsequent electron transfer takes place between the electron-deficient radical cation (NAP.+) and the electron-donor oxygen atom of zeolite framework. The aluminum rich MnZSM-5 zeolites (n = 3.4, 6.6) hinder efficiently the charge recombination and promote the electron transfer to generate a very long electron-hole pair which exceeds several weeks at room temperature in NAP@Li(6.6)ZSM-5. The electron-hole pair exhibits broad visible absorption bands at 482 and 525 nm. The electron-hole distance, 1.3 nm, was deduced from the dipolar interaction term (D) value. The spin density of trapped electron appears spread over (27)Al, (29)Si, (7)Li, and (1)H nuclei as deduced by two-dimensional approach of hyperfine sublevel correlation (HYSCORE). The very low recombination rate by tunneling effect was found to be in agreement with the very low value (J approximately 0) of the magnetic exchange. The combined effects of tight fit between NAP size and straight-channel dimension, the high aluminum content of the framework, and the highly polarizing cation Li(+) trapped efficiently the ejected electron in the conduction band and the hole in the valence band of the porous materials. PMID- 16262407 TI - Raman and IR spectroscopy of chemically processed single-walled carbon nanotubes. AB - IR and Raman spectroscopy has been used to study the evolution of the vibrational spectrum of bundled single-walled carbon nanotubes (SWNTs) during the purification process needed to remove metal catalyst and amorphous carbon present in arc-derived SWNT soot. We have carried out a systematic study to define the different outcomes stemming from the purification protocol (e.g., DO, DO/HCl, DO/HNO(3), H(2)O(2), H(2)O(2)/HCl), where dry oxidation (DO) or refluxing in H(2)O(2) was used in a first purification step to remove amorphous carbon. The second step involves acid reflux (HCl or HNO(3)) to remove the residual growth catalyst (Ni-Y). During strong chemical processing, it appears possible to create additional defects where carbon atoms are eliminated, the ring structure is now open, localized C=C bonds are created, and O-containing groups can be added to this defect to stabilize the structure. Evolution of SWNT skeletal disorder obtained via chemical processing was studied by Raman scattering. Higher intensity ratios of R- and G-band (I(R)/I(G)) are more typically found in SWNT materials with low D-band intensity and narrow G-band components. Using IR transmission through thin films of nanotubes, we can resolve the structure due to functional groups that were present in the starting material or added through chemical processing. After high-temperature vacuum annealing of the purified material at 1100 degrees C, IR spectroscopy shows that most of the added functional groups can be removed and that the structure that remains is assigned to the one- and two-phonon modes of SWNTs. PMID- 16262408 TI - Adsorption of ions to the surface of dilute electrolyte solutions: the Jones-Ray effect revisited. AB - The controversial observation of a minimum in the surface tension of dilute aqueous electrolyte solutions by Jones and Ray in the 1930s is confirmed by new resonance-enhanced second harmonic generation (SHG) experiments demonstrating surface enhancement of simple inorganic anions in the same concentration range. New experiments show that the quadruply charged ferrocyanide, Fe(CN)(6)(4-), anion is not surface active at high concentrations, as expected, but at dilute concentrations, the anion is strongly attracted to the interface with a Gibbs free energy of adsorption of -6.8 kcal/mol. Using this value, the original Jones and Ray data are fit to a simple model of the surface tension with qualitative agreement, although better agreement is found for all 13 Jones and Ray salts with an even stronger surface adsorption. PMID- 16262406 TI - New hints on the Ph-driven tautomeric equilibria of the topotecan anticancer drug in aqueous solutions from an integrated spectroscopic and quantum-mechanical approach. AB - The equilibria between the different forms of the topotecan anticancer drug have been studied at moderately acidic and physiological pH by an integrated computational tool rooted in the density functional theory and its time-dependent extension together with the polarizable continuum model. The results allow an unbiased selection between the different possible tautomeric forms and provide invaluable complements to experimental data. The ultraviolet-visible topotecan spectrum, recorded at moderately acidic pH, is accurately reproduced only by TD DFT computations including solvent effects. Comparison of the experimental and calculated bands of the UV-vis spectrum at physiological pH indicates the presence of an equilibrium among different forms that is tuned by the microenvironment embedding the drug. The quantitative agreement between TD DFT/PCM computations and experiments allows the identification of unequivocal spectroscopic signatures for different forms of topotecan. PMID- 16262410 TI - The peculiar trend of cyclic perfluoroalkane electron affinities with increasing ring size. AB - The adiabatic electron affinities (AEAs), vertical electron affinities (VEAs), and vertical detachment energies (VDEs) of cyclic perfluoroalkanes, c-C(n)F(2n) (n = 3-7), and their monotrifluoromethyl derivatives were computed using various pure and hybrid density functionals with DZP++ (polarization and diffuse function augmented double-zeta) basis sets. The theoretical AEA of c-C(4)F(8) at KMLYP/DZP++ is 0.70 eV, which exhibits satisfactory agreement with the 0.63 +/- 0.05 eV experimental value. The nonzero-point-corrected AEA of c-C(4)F(8) is predicted to be 0.41 eV at the CCSD(T)/aug-cc-pVTZ//MP2/aug-cc-pVTZ level of theory, which shows a slight deviation of 0.11 eV from the KMLYP estimated value of 0.52 eV for the same. With the zero-point correction from the MP2/6-311G(d) [Gallup, G. A. Chem. Phys. Lett. 2004, 399, 206] level of theory combined with the CCSD(T)/aug-cc-pVTZ//MP2/aug-cc-pVTZ result, the most reliable estimate of AEA of c-C(4)F(8) is 0.60 eV. c-C(3)F(6)(-), c-C(4)F(8)(-), and c-C(5)F(10)(-) are unusual in preferring planar to near planar ring structures. The ZPE corrected AEAs of c-C(n)F(2n) increase from n = 3 (0.24 eV) to n = 5 (0.77 eV), but then dramatically fall off to 0.40 eV for both n = 6 and n = 7. All of the other functionals predict the same trend. This is due to a change in the structural preference: C(s)() c-C(6)F(12)(-) and C(1) c-C(7)F(14)(-) are predicted to favor nonplanar rings, each with an exceptionally long C-F bond. (There also is a second, higher energy D3d minimum for C(6)F(12)(-).) The SOMOs as well as the spin density plots of the c-PFA radical anions reveal that the "extra" electron is largely localized on the unique F atoms in the larger n = 6 and n = 7 rings but is delocalized in the multiatom SOMOs of the three- to five membered ring radical anions. The computed AEAs are much larger than the corresponding VEAs; the latter are not consistent with different functionals. The AEAs are substantially larger when a c-C(n)()F(2)(n)() fluorine is replaced by a CF(3) group. This behavior is general; PFAs with tertiary C-F bonds have large AEAs. The VDEs for all the anions are substantial, ranging from 1.89 to 3.64 eV at the KMLYP/DZP++ level. PMID- 16262409 TI - Al-catalyzed enantioselective alkylation of alpha-ketoesters by dialkylzinc reagents. enhancement of enantioselectivity and reactivity by an achiral Lewis base additive. AB - An Al-catalyzed enantioselective method for additions of Me(2)Zn and Et(2)Zn to alpha-ketoesters bearing aromatic, alkenyl, and alkyl substituents is disclosed. Transformations are promoted in the presence of a readily available amino acid based ligand and afford the desired products in excellent yields and in up to 95% ee. Investigations described illustrate that the presence of a Lewis basic additive can lead to significant enhancements in efficiency and enantioselectivity. A mechanistic model that provides a rationale for such effects is provided. PMID- 16262411 TI - Kinetic and spectroscopic studies of the [palladium(Ar-bian)]-catalyzed semi hydrogenation of 4-octyne. AB - The kinetics of the stereoselective semi-hydrogenation of 4-octyne in THF by the highly active catalyst [Pd{(m,m'-(CF(3))(2)C(6)H(3))-bian}(ma)] (2) (bian = bis(imino)acenaphthene; ma = maleic anhydride) has been investigated. The rate law under hydrogen-rich conditions is described by r = k[4 octyne](0.65)[Pd][H(2)], showing first order in palladium and dihydrogen and a broken order in substrate. Parahydrogen studies have shown that a pairwise transfer of hydrogen atoms occurs in the rate-limiting step. In agreement with recent theoretical results, the proposed mechanism consists of the consecutive steps: alkyne coordination, heterolytic dihydrogen activation (hydrogenolysis of one Pd-N bond), subsequent hydro-palladation of the alkyne, followed by addition of N-H to palladium, reductive coupling of vinyl and hydride and, finally, substitution of the product alkene by the alkyne substrate. Under hydrogen limiting conditions, side reactions occur, that is, formation of catalytically inactive palladacycles by oxidative alkyne coupling. Furthermore, it has been shown that (Z)-oct-4-ene is the primary reaction product, from which the minor product (E)-oct-4-ene is formed by an H(2)-assisted, palladium-catalyzed isomerization reaction. PMID- 16262412 TI - Perforated layer structures in liquid crystalline rod-coil block copolymers. AB - We report a novel observation of the tetragonal perforated layer structures in a series of rod-coil liquid crystalline block copolymers (BCPs), poly(styrene-block (2,5-bis[4-methoxyphenyl]oxycarbonyl)styrene) (PS-b-PMPCS). PMPCS forms rigid rods while PS forms the coil block. Differential scanning calorimetry (DSC), polarized light microscopy (PLM), small-angle X-ray scattering (SAXS), wide-angle X-ray diffraction (WAXD), and transmission electron microscopy (TEM) techniques were used to investigate these rod-coil molecules, and a perforated layer structure was observed at f(PMPCS) approximately 0.37 in relatively low molecular weight (M(w)) samples and approximately 0.5 in high M(w) PS-b-PMPCS. This substantial phase boundary shift was attributed to the rod-coil nature of the BCP. The perforation obeys a tetragonal instead of hexagonal symmetry. The "onset" of perforation was also observed in real space in sample PS(272)-b PMPCS(93) (f(PMPCS) approximately 0.52), in which few PS chains punctuate PMPCS layers. A slight increase in f(PS), by blending with PS homopolymer, led to a dramatic change in the BCP morphology, and uniform tetragonal perforations were observed at f(PMPCS) approximately 0.48. PMID- 16262413 TI - Reactant concentrations from fluorescence correlation spectroscopy with tailored fluorescent probes. An example of local calibration-free pH measurement. AB - The present account is concerned with the measurement of local reactant concentrations by observing specific fluorescent probes in fluorescence correlation spectroscopy (FCS). The Theoretical Analysis section revisits the photophysical, thermodynamic, and kinetic information that is contained in the corresponding FCS correlation curves. In particular, we examine the conditions under which FCS is revealed as a superior tool to measure concentrations of reactive species. Careful molecular engineering of the specific fluorescent probes that simultaneously integrates photophysical, thermodynamic, and kinetic constraints will be required to benefit most from FCS. We illustrate the FCS titration approach with a series of fluorescent probes that we tailored to measure pH at around 4-6 by FCS after two-photon excitation. We show that an optimal design allows one to access pH without any preliminary calibrations such as the determination of the protonation constant or the photophysical properties of the fluorescent probe. PMID- 16262414 TI - A simple iridium catalyst with a single resolved stereocenter for enantioselective allylic amination. Catalyst selection from mechanistic analysis. AB - A study of the relationship between the stereochemical elements of a phosphoramidite ligand and the stereoselectivity of iridium-catalyzed amination of allylic carbonates is reported. During catalyst activation, a complex of a phosphoramidite ligand possessing one axial chiral binaphtholate group and two resolved phenethyl substituents converts to a more reactive cyclometalated complex containing one distal chiral substituent at nitrogen, one substituent that becomes part of the metalacycle, and one unperturbed binaphtholate group. Systematic changes were made to the different stereochemical elements. Replacement of the distal chiral phenethyl substituent with a large achiral cycloalkyl group led to a catalyst that reacts with rates and enantioselectivities that are similar to those of the original catalyst with the phenethyl group. Studies of the reactions of diastereomeric ligands containing (R) or (S) binaphtholate groups on phosphorus, along with one (R)-phenethyl and one achiral cyclododecyl group on nitrogen, show that the complexes of the two diastereomeric ligands undergo cyclometalation at much different rates. To access both diastereomeric catalysts and to determine if the reaction can occur selectively with an even simpler ligand containing a phenethyl substituent at nitrogen as the only resolved stereochemical element, the catalyst derived from a phosphoramidite containing a biphenolate group was studied. Catalysts generated from this ligand were shown to react in all cases examined with nearly the same rates, regioselectivities, and enantioselectivities as catalysts derived from the original more elaborate ligand. The absolute stereochemistry of the product implies that the major enantiomer is formed from the (R(a),R(c))-atropisomer of the catalyst containing the biphenolate group. PMID- 16262415 TI - On the correlation between bond-length change and vibrational frequency shift in hydrogen-bonded complexes: a computational study of Y...HCl dimers (Y = N2, CO, BF). AB - The H-Cl bond-length change and the harmonic vibrational frequency shift of the H Cl stretch on formation of the linear isoelectronic Y...H-Cl complexes (Y = N(2), CO, BF) have been determined by ab initio computations at different levels of theory. These shifts are in agreement with predictions from a model based on perturbation theory and involving the first and second derivatives of the interaction energy with respect to displacement of the H-Cl bond length from its equilibrium value in the isolated monomer. At the highest level of theory, blue shifts were obtained for BF...HCl and CO...HCl, while red shifts were obtained for FB...HCl, OC...HCl, and N(2)...HCl. These vibrational characteristics are rationalized by considering the balance between the interaction energy derivatives obtained from the perturbative model. The widely believed correlation between the bond-length change and the sign of the frequency shift obtained on complexation is discussed and found to be unreliable. PMID- 16262416 TI - Substrate activity screening: a fragment-based method for the rapid identification of nonpeptidic protease inhibitors. AB - A new fragment-based method for the rapid development of novel and distinct classes of nonpeptidic protease inhibitors, Substrate Activity Screening (SAS), is described. This method consists of three steps: (1) a library of N-acyl aminocoumarins with diverse, low molecular weight N-acyl groups is screened to identify protease substrates using a simple fluorescence-based assay, (2) the identified N-acyl aminocoumarin substrates are optimized by rapid analogue synthesis and evaluation, and (3) the optimized substrates are converted to inhibitors by direct replacement of the aminocoumarin with known mechanism-based pharmacophores. The SAS method was successfully applied to the cysteine protease cathepsin S, which is implicated in autoimmune diseases. Multiple distinct classes of nonpeptidic substrates were identified upon screening an N-acyl aminocoumarin library. Two of the nonpeptidic substrate classes were optimized to substrates with >8000-fold improvements in cleavage efficiency for each class. Select nonpeptidic substrates were then directly converted to low molecular weight, novel aldehyde inhibitors with nanomolar affinity to cathepsin S. This study demonstrates the unique characteristics and merits of this first substrate based method for the rapid identification and optimization of weak fragments and provides the framework for the development of completely nonpeptidic inhibitors to many different proteases. PMID- 16262417 TI - Chiral zirconium catalysts using multidentate BINOL derivatives for catalytic enantioselective Mannich-type reactions; ligand optimization and approaches to elucidation of the catalyst structure. AB - Catalytic enantioselective Mannich-type reactions of silicon enolates with aldimines were investigated using chiral zirconium catalysts prepared from Zr(O(t)Bu)(4), N-methylimidazole, and newly designed multidentate BINOL derivatives. These new multidentate BINOL ligands were designed on the basis of an assumed transition state structure of a chiral zirconium catalyst derived from two molecules of (R)-6,6'-Br(2)-BINOL. Not only tetradentate BINOL 4 but also tridentate BINOL derivatives were found to be effective, and high enantioselectivities were attained. In a structural study of the most effective zirconium complex prepared from tridentate ligand 6e, several NMR experiments and DFT calculations were carried out. Consequently, the structure of an active catalyst and plausible mechanism of asymmetric induction were elucidated. PMID- 16262418 TI - Large optical activity of gold nanocluster enantiomers induced by a pair of optically active penicillamines. AB - We have succeeded for the first time in preparing a pair of gold nanocluster enantiomers protected by optically active thiols: D- and L-penicillamine (D-Pen and L-Pen). Circular dichroism (CD) spectroscopy confirmed the mirror image relationship between the D-Pen-capped and the L-Pen-capped gold nanoclusters, suggesting that the surface modifier acts as a chiral selector, and that the nanoclusters have well-defined stereostructures as common chiral molecules do. No CD signals could be obtained when the gold nanoclusters were synthesized by using a racemic mixture (rac-Pen). These chiroptical properties were investigated for the three separated fractions of each of the gold nanoclusters (D-Pen-capped, L Pen-capped, or rac-Pen-capped clusters) by polyacrylamide gel electrophoresis (PAGE). Each fractioned component has the mean diameter of 0.57, 1.18, or 1.75 nm that was determined by a solution-phase small-angle X-ray scattering. With a decrease in the mean cluster diameter, optical activity or anisotropy factors gradually increased. On the basis of the kinetic and the structural considerations, the origins of large optical activity of the gold nanocluster enantiomers are discussed. PMID- 16262419 TI - Control of photochemical, photophysical, electrochemical, and photocatalytic properties of rhenium(I) complexes using intramolecular weak interactions between ligands. AB - Intramolecular interactions between ligands have been successfully applied as a novel tool for controlling various properties of a series of cis,trans [Re(dmb)(CO)(2)(PR(3))(PR'(3))](+)-type complexes (dmb = 4,4'-dimethyl-2,2' bipyridine), in the ground state and in the excited state and in the one-electron reduced form. For rhenium complexes with two triarylphosphine ligands, P(p XPh)(3), the dmb ligand was sandwiched by four aryl rings having CH(aryl) pi(pyridine)-pi(aryl) interactions. On the other hand, complexes with one triarylphosphine ligand and one trialkylphosphite ligand, P(OR)(3), had pi-pi and CH-pi interactions between each pyridine ring in the dmb ligand and the aryl group in the P(p-XPh)(3). Various properties of these two series of rhenium complexes were compared with those of complexes having two trialkylphosphite ligands, which do not interact through space with the dmb ligand. Properties of the complexes associated mainly with the dmb ligand are strongly affected by the intramolecular interactions: (1) UV/vis absorptions to the pi-pi and (1)MLCT excited states were both red-shifted, but (2) emission from the (3)MLCT excited state was blue-shifted; (3) the lifetime of the (3)MLCT excited state was prolonged up to 3-fold; (4) the reduction potential in the ground state was positively shifted by 110 mV with pi-pi and CH-pi interactions and by 180-200 mV with the CH-pi-pi interactions. (5) In the excited states, the oxidation power of the complex was also enhanced by the intramolecular interactions. (6) In the corresponding one-electron-reduced species cis,trans-[Re(dmb( .)(CO)(2)(PR(3))(PR'(3))], the intramolecular interactions are maintained and strongly affected their UV/vis spectra. (7) Photocatalysis for CO(2) reduction was significantly enhanced only by the CH-pi-pi interaction. PMID- 16262420 TI - Planar polarized light emission from CdSe nanoparticle clusters. AB - This paper describes synthesis and optical properties of planar clusters of CdSe nanocrystals. The clusters emit linearly polarized light in the plane of the cluster. The emission wavelength of the clusters can be adjusted between 568 and 639 nm with the size of the CdSe nanocrystals. Planar CdSe microclusters were synthesized by reaction of trioctylphosphine oxide-coated CdSe/CdS nanocrystals with 3-aminopropylsilyl-modified Ca(2)Nb(3)O(10) nanosheets in THF. The clusters are 3.92 +/- 1.18 mum length/width and 91 +/- 37 nm thickness, and they consist of alternating layers of Ca(2)Nb(3)O(10) to which CdSe nanocrystals are attached with densities of 5300 +/-310 particles per side of a single Ca(2)Nb(3)O(10) sheet. The chemical inertness of the clusters in coordinating solvents suggests covalent interactions between the aminopropyl groups and CdSe nanocrystals. Upon excitation at lambda(exc) = 400 nm, the clusters emit green (568 nm), orange (589 nm), or red (639 nm) light, depending on the size of the CdSe crystals. The light is emitted preferentially in the cluster plane and it is linearly polarized along the cluster edges. Combined fluorescence microscopy and atomic force microscopy reveal that the directional emission efficiency depends linearly on the thickness of the clusters, which varies between 70 and 180 nm. The ability to manipulate the direction and polarization of the photoemission of CdSe nanoparticles via assembly into 2D structures is of interest for applications of these and similar structures in advanced optical materials and devices. PMID- 16262421 TI - Electrical discharge in a nanometer-sized air/water gap observed by atomic force microscopy. AB - We report a method to initiate and investigate electrical discharges of ambient air/water molecules in a nanometer-sized gap. Our methodology is based on a typical atomic force microscopy (AFM) setup, in which a cylinder discharge gap of < or =5 nm could be configured between the AFM probe and substrate. We observed highly localized stochastic nanoexplosions in which the discharge probability is dominated by the electric field, material-specific surface reactions, and humidity. AFM results, coupled with the boundary element method (BEM), finite element method (FEM), and method of characteristics (MOC) simulations, further revealed the generation of transient shock waves in the nanoscale discharge. The propagation of shock fronts significantly facilitates the radial expansion of the ionized particles, leading to the formation of microscale patterns on selected substrates. Our findings provide an initial understanding of nanoscale discharge and could be relevant to a few applications including nano/microstructuring, microelectronics, and plasma-assisted depositions. PMID- 16262422 TI - Activation of high-TC ferromagnetism in Co2+:TiO2 and Cr3+:TiO2 nanorods and nanocrystals by grain boundary defects. AB - Colloidal Co(2+)- and Cr(3+)-doped TiO(2) nanorods and nanocrystals were synthesized and studied by X-ray powder diffraction, electronic absorption spectroscopy, magnetic circular dichroism spectroscopy, magnetic susceptibility, and transmission electron microscopy. The nanorods were paramagnetic as colloids but showed room-temperature ferromagnetism when spin-coated aerobically into films. Crystalline domain size, thermal annealing, and dopant or defect migration are not the dominating factors converting the doped TiO(2) nanocrystals from the paramagnetic state to the ferromagnetic state. The most important factor for activating ferromagnetism is found to be the creation of grain boundary defects, proposed to be oxygen vacancies at nanocrystal fusion interfaces. These defects are passivated and the ferromagnetism destroyed by further aerobic annealing. These results not only help elucidate the origins of the TM(n+):TiO(2) DMS ferromagnetism but also represent an advance toward the controlled manipulation of high-T(C) DMS ferromagnetism using external chemical perturbations. PMID- 16262424 TI - Carbon-fluorine bond activation coupled with carbon-hydrogen bond formation alpha to iridium: kinetics, mechanism, and diastereoselectivity. AB - Reactions of iridium(fluoroalkyl)hydride complexes CpIr(PMe(3))(CF(2)R(F))Y (R(F) = F, CF(3); Y = H, D) with LutHX (Lut = 2,6-dimethylpyridine; X = Cl, I) results in C-F activation coupled with hydride migration to give CpIr(PMe(3))(CYFR(F))X as variable mixtures of diastereomers. Solution conformations and relative diastereomer configurations of the products have been determined by (19)F{(1)H}HOESY NMR to be (S(C), S(Ir))(R(C), R(Ir)) for the kinetic diastereomer and (R(C), S(Ir))(S(C), R(Ir)) for its thermodynamic counterpart. Isotope labeling experiments using LutDCl/CpIr(PMe(3))(CF(2)R(F))H and CpIr(PMe(3))(CF(2)R(F))D/LutHCl) showed that, unlike a previously studied system, H/D exchange is faster than protonation of the alpha-CF bond, giving an identical mixture of product isotopologues from both reaction mixtures. The kinetic rate law shows a first-order dependence on the concentration of iridium substrate, but a half-order dependence on that of LutHCl; this is interpreted to mean that LutHCl dissociates to give HCl as the active protic source for C-F bond activation. Detailed kinetic studies are reported, which demonstrate that lack of complete diastereoselectivity is not a function of the C-F bond activation/H migration steps but that a cationic intermediate plays a double role in loss of diastereoselectivity; the intermediate can undergo epimerization at iridium before being trapped by halide and can also catalyze the epimerization of kinetic diastereomer product to thermodynamic product. A detailed mechanism is proposed and simulations performed to fit the kinetic data. PMID- 16262423 TI - Effect of temperature, cholesterol content, and antioxidant structure on the mobility of vitamin E constituents in biomembrane models studied by laterally diffusion-controlled fluorescence quenching. AB - Kinetic parameters relevant for the antioxidant activity of the vitamin E constituents (alpha, beta, gamma, and delta homologues of tocopherols and tocotrienols) and of an amphiphilic vitamin C derivative, l-ascorbyl 6-palmitate, were determined. Fluorescence quenching experiments of 2,3-diazabicyclo[2.2.2]oct 2-ene in homogeneous acetonitrile-water mixtures afforded reactivity trends in terms of intermolecular quenching rate constants, while the quenching of Fluorazophore-L in liposomes provided the lateral diffusion coefficients relevant for understanding their biological activity in membranes. The reactivity in homogeneous solution was not influenced by the nature of the isoprenoid tail (tocopherol versus tocotrienol), but was dependent on the methylation pattern. The resulting order (alpha > beta = gamma > delta) was found to be in line with their reactivities toward peroxyl radicals as well as the phenolic O-H bond dissociation energies. The mutual lateral diffusion coefficient in POPC liposomes was the same, within error, for different tocopherols and tocotrienols (D(L) = (1.6 +/- 0.2) x 10(-7) cm(2) s(-1)). l-Ascorbyl 6-palmitate exhibited a reactivity similar to that of delta-tocopherol in homogeneous solution, but displayed a 1 order of magnitude lower fluorescence quenching efficiency in liposomes than the vitamin E constituents. Temperature effects on the laterally diffusion-controlled fluorescence quenching were large, with activation energies of 44 +/- 6 kJ mol(-1). The addition of cholesterol (0-30%) to POPC liposomes resulted only in slightly reduced diffusion coefficients. The combined results demonstrate that Fluorazophore-L can provide important physicochemical parameters for the understanding of antioxidant activity in biological environments. PMID- 16262425 TI - Growth and assembly of crystalline tungsten oxide nanostructures assisted by bioligation. AB - Here, we present a systematic study on the influence of the bioligand deferoxamine mesylate on the crystallization and assembly behavior of tungsten oxide in a soft-chemistry process. Without deferoxamine mesylate, this approach yields pseudo-single crystalline tungstite nanoplatelets consisting of a large number of crystallographically almost perfectly aligned primary crystallites. In the presence of a constant amount of deferoxamine, the particle morphology drastically changes with temperature, ranging from wormlike organic-inorganic hybrid nanostructures to single-crystalline tungsten oxide nanowires, highlighting the role of the bioligand in controlling the crystal growth and assembly behavior. The nanowires have a uniform diameter of about 1.3 nm, an aspect ratio of more than 500, and the structural flexibility of tungsten oxide. The presented process is based on the combination of biomimetic construction principles with nonaqueous sol-gel chemistry, thus combining the advantages of both tools, excellent control over particle morphology and high crystallinity at low temperature. PMID- 16262426 TI - Multiple-site exchange in proteins studied with a suite of six NMR relaxation dispersion experiments: an application to the folding of a Fyn SH3 domain mutant. AB - The three-site exchange folding reaction of an (15)N-labeled, highly deuterated Gly48Met mutant of the Fyn SH3 domain has been characterized at 25 degrees C using a suite of six CPMG-type relaxation dispersion experiments that measure exchange contributions to backbone (1)H and (15)N transverse relaxation rates in proteins. It is shown that this suite of experiments allows the extraction of all the parameters of this multisite exchange process in a robust manner, including chemical shift differences between exchanging states, from a data set recorded at only a single temperature. The populations of the exchanging folded, intermediate, and unfolded states that are fit are 94, 0.7, and 5%, respectively. Despite the small fraction of the intermediate, structural information is obtained for this state that is consistent with the picture of SH3 domain folding that has emerged from other studies. Taken together, the six dispersion experiments facilitate the complete reconstruction of (1)H-(15)N correlation spectra for the unfolded and intermediate states that are "invisible" in even the most sensitive of NMR experiments. PMID- 16262427 TI - Differential solvation and tautomer stability of a model base pair within the minor and major grooves of DNA. AB - 2-(2'-Hydroxyphenyl)benzoxazole (HBO) may be used as a model base pair to study solvation, duplex environment, and tautomerization within the major and minor groves of DNA duplexes. In its ground state, HBO possesses an enol moiety which may be oriented syn or anti relative to the imino nitrogen of the benzoxazole ring. In the absence of external hydrogen-bond donors and acceptors HBO exists as the internally hydrogen-bonded syn-enol, a mimic of the rare base pair tautomer found in DNA, which may be photoinduced to tautomerize and form the keto tautomer, a mimic of the dominant base pair tautomer. Previously, we demonstrated that when incorporated into DNA such that the enol moiety is positioned in the major groove, HBO is not solvated, exists exclusively as the internally hydrogen bonded syn-enol which is efficiently photoinduced to tautomerize, and the corresponding keto tautomer is preferentially stabilized. In stark contrast, we now show that when HBO is incorporated in DNA such that the enol moiety is positioned in the minor groove, the enol tautomer is preferentially stabilized. Molecular dynamics simulations suggest that this results from the formation of a stable hydrogen-bond between the HBO enol and the O4' atom of an adjacent nucleotide, an H-bond acceptor that is only available in the minor groove. The differential stabilization of the enol and keto tautomers in the major and minor grooves may reflect the functions for which these environments evolved, including duplex replication, stability, and recognition. PMID- 16262428 TI - Nano-segregated polymeric film exhibiting high ionic conductivities. AB - Nanostructures can be used for the fabrication of highly functional materials transporting ions and charges. We demonstrate a new design strategy for polymeric higher ion-conductors. Phase-segregated layers of alternating mobile tetra(ethylene oxide)s (TEOs) and rigid aromatic cores where the TEO moieties are grafted from aromatic layers have been shown to be efficient to transport lithium triflate. Such segregated structures at the nanometer scale (nano-segregated structures) were prepared by in-situ photopolymerization of an aligned methacrylate liquid crystalline monomer comprising a terphenyl rigid rod mesogen having a TEO terminal chain. The ion-conductive TEO moiety remains in the highly mobile state even after polymerization, which is indicated by its low glass transition temperature (-45 degrees C). This nanostructured film exhibits an ionic conductivity parallel to the layer of 10(-3) S cm(-1) at room temperature. The highest ionic conductivity is in the level of 10(-2) S cm(-1) observed at 150 degrees C. The anisotropic ionic conductivities have been observed for the nano segregated film. PMID- 16262429 TI - Photocontrol of DNA binding specificity of a miniature engrailed homeodomain. AB - Control of DNA binding of HDH-3, a 18-residue polypeptide based on the recognition helix of the Q50K engrailed homeodomain, has been achieved. HDH-3 was linked to an azobenzene cross-linker through two cysteine residues in an i, i + 11 spacing. For the thermodynamically stable trans configuration of the cross linker, the dark-adapted peptide (dad-HDH-3) adopted a mainly alpha-helical structure as judged by circular dichroism (CD) spectroscopy. After irradiation with light of 360 nm, the helical content of the peptide (irrad-HDH-3) was reduced significantly and the CD spectrum of the irradiated peptide resembled that of the largely unstructured, unalkylated peptide. Despite lacking helices-1 and -2 and the N-terminal arm of Q50K engrailed, dad-HDH-3 bound to its natural DNA target sequence TAATCC (QRE) with high affinity (K(D) = 7.5 +/- 1.3 nM). The binding affinity for the mutant DNA sequence, TAATTA (ERE), was reduced significantly (K(D) = 140 +/- 11 nM). Unlike irrad-HDH-3, which like the unalkylated parent peptide displayed only marginal DNA binding specificity, dad HDH-3 specified base pairs 5 and 6 of QRE with an accuracy rivaling that of the intact wild-type Q50K engrailed homeodomain, making dad-HDH-3 the most specific designed DNA binding miniature homeodomain reported to date. Moreover, DNA binding affinity and specificity of HDH-3 could be controlled externally by irradiation with light. PMID- 16262430 TI - Direct observation of a transverse vibrational mechanism for negative thermal expansion in Zn(CN)2: an atomic pair distribution function analysis. AB - The instantaneous structure of the cyanide-bridged negative thermal expansion (NTE) material Zn(CN)(2) has been probed using atomic pair distribution function (PDF) analysis of high energy X-ray scattering data (100-400 K). The temperature dependence of the atomic separations extracted from the PDFs indicates an increase of the average transverse displacement of the cyanide bridge from the line connecting the Zn(II) centers with increasing temperature. This allows the contraction of non-nearest-neighbor Zn...Zn' and Zn...C/N distances despite the observed expansion of the individual direct Zn-C/N and C-N bonds. Thus, this analysis provides definitive structural confirmation that an increase in the average displacement of bridging atoms is the origin of the NTE behavior. The lattice parameters reveal a slight reduction in the NTE behavior at high temperature from a minimum coefficient of thermal expansion (alpha = dl/ldT) of 19.8 x 10(-6) K(-1) below 180 K, which is attributed to interaction between the doubly interpenetrated frameworks that comprise the structure. PMID- 16262431 TI - Nonequilibrium synthesis and assembly of hybrid inorganic-protein nanostructures using an engineered DNA binding protein. AB - We show that a protein with no intrinsic inorganic synthesis activity can be endowed with the ability to control the formation of inorganic nanostructures under thermodynamically unfavorable (nonequilibrium) conditions, reproducing a key feature of biological hard-tissue growth and assembly. The nonequilibrium synthesis of Cu(2)O nanoparticles is accomplished using an engineered derivative of the DNA-binding protein TraI in a room-temperature precursor electrolyte. The functional TraI derivative (TraIi1753::CN225) is engineered to possess a cysteine constrained 12-residue Cu(2)O binding sequence, designated CN225, that is inserted into a permissive site in TraI. When TraIi1753::CN225 is included in the precursor electrolyte, stable Cu(2)O nanoparticles form, even though the concentrations of [Cu(+)] and [OH(-)] are at 5% of the solubility product (K(sp,Cu2O)). Negative control experiments verify that Cu(2)O formation is controlled by inclusion of the CN225 binding sequence. Transmission electron microscopy and electron diffraction reveal a core-shell structure for the nonequilibrium nanoparticles: a 2 nm Cu(2)O core is surrounded by an adsorbed protein shell. Quantitative protein adsorption studies show that the unexpected stability of Cu(2)O is imparted by the nanomolar surface binding affinity of TraIi1753::CN225 for Cu(2)O (K(d) = 1.2 x 10(-)(8) M), which provides favorable interfacial energetics (-45 kJ/mol) for the core-shell configuration. The protein shell retains the DNA-binding traits of TraI, as evidenced by the spontaneous organization of nanoparticles onto circular double-stranded DNA. PMID- 16262432 TI - Total synthesis of the thiopeptide antibiotic amythiamicin D. AB - The thiopeptide (or thiostrepton) antibiotics are a class of sulfur containing highly modified cyclic peptides with interesting biological properties, including reported activity against MRSA and malaria. Described herein is the total synthesis of the thiopeptide natural product amythiamicin D, which utilizes a biosynthesis-inspired hetero-Diels-Alder route to the pyridine core of the antibiotic as a key step. Preliminary studies using a range of serine-derived 1 ethoxy-2-azadienes established that hetero-Diels-Alder reaction with N acetylenamines proceeded efficiently under microwave irradiation to give 2,3,6 trisubstituted pyridines. The thiazole building blocks of the antibiotic were obtained by either classical Hantzsch reactions or by dirhodium(II)-catalyzed chemoselective carbene N-H insertion followed by thionation, and were combined to give the bis-thiazole that forms the left-hand fragment of the antibiotic. The key Diels-Alder reaction of a tris-thiazolyl azadiene with benzyl 2-(1 acetylaminoethenyl)thiazole-4-carboxylate gave the core tetrathiazolyl pyridine, which was elaborated into the natural product by successive incorporation of glycine and bis-thiazole fragments followed by macrocyclization. PMID- 16262433 TI - Total synthesis of (-)-crambidine and definition of the relative configuration of its unique tetracyclic guanidinium core. AB - Total syntheses of the 3S,8S,10S,19R,43S isomer 4a and the 3S,8S,10S,19R,43R isomer 4b of the unique crambescidin alkaloid crambidine are reported. These studies confirm the tetracyclic structure proposed by Braekman and co-workers for crambidine, and establish the rel-3R,8R,10R,19S relative configuration for this moiety. Natural crambidine is most likely the 3S,8S,10S,19R,43S isomer 4a. These syntheses were completed in five steps and approximately 14% overall yield from 1 iminohexahydro[1,2-c]pyrimidine carboxylic ester 10, an intermediate in our earlier total synthesis of 13,14,15-isocrambescidin 800 (3). The signature step in the total syntheses of crambidine and several stereoisomers is chemoselective dehydrogenation of the tethered Biginelli adduct 10 or the derived tetracyclic intermediate 17. Additionally, these studies reveal the unprecedented ring-chain isomerization of the crambidine ring system exemplified by the interconversion of isomers 15a and 15b. PMID- 16262434 TI - Web resources: the vanishing act. PMID- 16262435 TI - American Dental Association white paper targets dental care for the underserved. PMID- 16262436 TI - Evidence-based practice and school nursing. AB - School nurses need to demonstrate that their practice is based on the best evidence available, which is usually data obtained from research. Evidence-based practice involves combining the best evidence available with nursing expertise and patient and family preferences to determine optimum care. Evidence-based practice guidelines are developed by carefully reviewing the available evidence on a topic and synthesizing this information into recommendations for practice. This article defines evidence-based practice and best evidence, describes the development of evidence-based practice guidelines, discusses factors that affect the use of research and evidence-based practice guidelines in school nursing, and reviews current sources of evidence-based practice guidelines for school nurses. Strategies that school nurses can use to incorporate evidence into their practice are discussed. One recommendation is that school nurses partner with nurse leaders and nurse researchers to develop evidence-based practice guidelines relevant to school nurse practice. PMID- 16262437 TI - An oral health education program for Latino immigrant parents. AB - A high prevalence of dental caries in the pediatric population is a major health problem. At highest risk are low-income minority groups, including refugee and immigrant populations. Consequences of oral disease include pain, difficulty eating and speaking, poor school performance, and poor self-esteem. Parent involvement in oral health education is crucial. This program provided oral health education for Latino immigrant parents in a northern California school district. A pretest-posttest was administered to measure changes in oral health knowledge and reported oral health behaviors following two sessions of oral health education. This program provides a framework for school nurses who are in an ideal position to implement similar programs that address the oral health needs of the pediatric population, particularly those of the Latino immigrant community. PMID- 16262438 TI - Rampant caries. AB - Although dental caries in the pediatric and adolescent population has consistently declined in the United States, it is still the most common childhood disease. Dental problems are the number one reason for missing school next to the common cold. Dental caries are an infectious, communicable disease resulting in destruction of tooth structure by acid-forming bacteria found in dental plaque, an intraoral biofilm, in the presence of sugar. The etiology of rampant disease is very complex. It is multifactorial and has a close relationship to a number of risk factors. It is important that school nurses identify patients at risk for dental caries early to determine their risk and refer them for disease prevention and control. PMID- 16262439 TI - Tooth avulsion in the school setting. AB - Tooth avulsions occur when a tooth is displaced from its socket. Tooth avulsions are common dental injuries that may occur before, during, or after school. Therefore, it is essential that school nurses be well prepared to intervene when such a dental emergency arises. It is also imperative that school nurses and school personnel are fully equipped to manage a tooth avulsion. Time is of the essence when attempting to save an avulsed tooth. The goal is to successfully reinsert the avulsed tooth and to preserve the periodontal ligament. The school nurse is responsible for implementing the appropriate actions needed to save the avulsed tooth. PMID- 16262440 TI - Assessing the capability of school-age children with asthma to safely self-carry an inhaler. AB - The purpose of this descriptive study was to pilot test an Asthma Assessment Interview (AAI) and to determine the approximate age a child with asthma is capable to self-carry an inhaler. A random sample of 34 students with asthma (Grades K through 10) from a midwestern school district were interviewed by the school nurse using the AAI, which assesses knowledge of asthma, symptoms, coping strategies, medication administration skills, triggers, and judgment about when to use an inhaler including the ability to tell time. Only 38% passed the AAI. No students ages 5 to 7 passed, fewer than 50% of students ages 8 to 10 passed, and half or more of students age 11 or older passed the AAI. Results suggest a school nurse should supervise elementary students when using an inhaler; most should not self-carry. The AAI can be a useful part of the school nurse's assessment. PMID- 16262441 TI - The relevance of standards of professional school nursing practice. AB - This descriptive correlational study assessed school nurses' knowledge of and perceived relevance of the Standards of Professional School Nursing Practice. Of the 1,162 Ohio school nurses sent questionnaires, 345 returned usable questionnaires (30%). The typical respondent was a 50-year-old Caucasian woman with 24 years of nursing experience, 12 years of school nursing experience, and a caseload of 3 public schools serving 1,500 students. Factors that positively related to the school nurses' knowledge and perceptions of the relevance of the standards were age, advanced degree preparation, school nursing experience, and practicing in a metropolitan area. Factors that negatively related to knowledge and perceived relevance were practicing in a rural area and caseload of special needs students. Standards of practice provide a mechanism for defining and supporting practice roles for nurses. Thus, it is important to increase awareness and use of the standards, especially among school nurses practicing in rural areas. PMID- 16262442 TI - Oral health screening using a caries detection device. AB - Children from low-income families are more likely to have dental disease and less likely to have regular dental care. Children's dental-related illnesses are responsible for more than 880,000 lost school days each year. The lack of reliable state-level data often makes monitoring the oral health of children a challenge. By conducting oral health screenings, school nurses could promote prevention and early identification of dental problems and could aid in the collection of data. This innovative project used a convenience sample of students to test a caries detection device. Children in Grades 3 and 6 (n = 2,622) received oral health screening by the school nurse using a noninvasive laser fluorescent dental device. Findings revealed that 82% of students referred to the dentists received dental care. School nurses can contribute to improving the oral health status of the school-age population by early detection of dental caries and referrals for a dental evaluation. PMID- 16262444 TI - Different catalytic mechanisms in mammalian selenocysteine- and cysteine containing methionine-R-sulfoxide reductases. AB - Selenocysteine (Sec) is found in active sites of several oxidoreductases in which this residue is essential for catalytic activity. However, many selenoproteins have fully functional orthologs, wherein cysteine (Cys) occupies the position of Sec. The reason why some enzymes evolve into selenoproteins if the Cys versions may be sufficient is not understood. Among three mammalian methionine-R-sulfoxide reductases (MsrBs), MsrB1 is a Sec-containing protein, whereas MsrB2 and MsrB3 contain Cys in the active site, making these enzymes an excellent system for addressing the question of why Sec is used in biological systems. In this study, we found that residues, which are uniquely conserved in Cys-containing MsrBs and which are critical for enzyme activity in MsrB2 and MsrB3, were not required for MsrB1, but increased the activity of its Cys mutant. Conversely, selenoprotein MsrB1 had a unique resolving Cys reversibly engaged in the selenenylsulfide bond. However, this Cys was not necessary for activities of either MsrB2, MsrB3, or the Cys mutant of MsrB1. We prepared Sec-containing forms of MsrB2 and MsrB3 and found that they were more than 100-fold more active than the natural Cys forms. However, these selenoproteins could not be reduced by the physiological electron donor, thioredoxin. Yet, insertion of the resolving Cys, which was conserved in MsrB1, into the selenoprotein form of MsrB3 restored the thioredoxin-dependent activity of this enzyme. These data revealed differences in catalytic mechanisms between selenoprotein MsrB1 and non-selenoproteins MsrB2 and MsrB3, and identified catalytic advantages and disadvantages of Sec- and Cys-containing proteins. The data also suggested that Sec- and Cys-containing oxidoreductases require distinct sets of active-site features that maximize their catalytic efficiencies and provide strategies for protein design with improved catalytic properties. PMID- 16262445 TI - Mechanism of filament nucleation and branch stability revealed by the structure of the Arp2/3 complex at actin branch junctions. AB - Actin branch junctions are conserved cytoskeletal elements critical for the generation of protrusive force during actin polymerization-driven cellular motility. Assembly of actin branch junctions requires the Arp2/3 complex, upon activation, to initiate a new actin (daughter) filament branch from the side of an existing (mother) filament, leading to the formation of a dendritic actin network with the fast growing (barbed) ends facing the direction of movement. Using genetic labeling and electron microscopy, we have determined the structural organization of actin branch junctions assembled in vitro with 1-nm precision. We show here that the activators of the Arp2/3 complex, except cortactin, dissociate after branch formation. The Arp2/3 complex associates with the mother filament through a comprehensive network of interactions, with the long axis of the complex aligned nearly perpendicular to the mother filament. The actin-related proteins, Arp2 and Arp3, are positioned with their barbed ends facing the direction of daughter filament growth. This subunit map brings direct structural insights into the mechanism of assembly and mechanical stability of actin branch junctions. PMID- 16262451 TI - Introduction to the special section on developing guidelines for the evidence based assessment (EBA) of adult disorders. AB - The goal of this special section is to encourage greater awareness of evidence based assessment (EBA) in the development of a scientifically supported clinical psychology. In this introductory article, the authors describe the elements that authors in this special section were asked to consider in their focused reviews (including the scope of available psychometric evidence, advancements in psychopathology research, and evidence of attention to factors such as gender, age, and ethnicity in measure validation). The authors then present central issues evident in the articles that deal with anxiety, depression, personality disorders, and couple distress and in the accompanying commentaries. The authors conclude by presenting key themes emerging from the articles in this special section, including gaps in psychometric information, limited information about the utility of assessment, the discrepancy between recommended EBAs and current training and practice, and the need for further data on the process of clinical assessment. PMID- 16262446 TI - Dynamic acetylation of all lysine 4-methylated histone H3 in the mouse nucleus: analysis at c-fos and c-jun. AB - A major focus of current research into gene induction relates to chromatin and nucleosomal regulation, especially the significance of multiple histone modifications such as phosphorylation, acetylation, and methylation during this process. We have discovered a novel physiological characteristic of all lysine 4 (K4)-methylated histone H3 in the mouse nucleus, distinguishing it from lysine 9 methylated H3. K4-methylated histone H3 is subject to continuous dynamic turnover of acetylation, whereas lysine 9-methylated H3 is not. We have previously reported dynamic histone H3 phosphorylation and acetylation as a key characteristic of the inducible proto-oncogenes c-fos and c-jun. We show here that dynamically acetylated histone H3 at these genes is also K4-methylated. Although all three modifications are proven to co-exist on the same nucleosome at these genes, phosphorylation and acetylation appear transiently during gene induction, whereas K4 methylation remains detectable throughout this process. Finally, we address the functional significance of the turnover of histone acetylation on the process of gene induction. We find that inhibition of turnover, despite causing enhanced histone acetylation at these genes, produces immediate inhibition of gene induction. These data show that all K4-methylated histone H3 is subject to the continuous action of HATs and HDACs, and indicates that at c-fos and c-jun, contrary to the predominant model, turnover and not stably enhanced acetylation is relevant for efficient gene induction. PMID- 16262450 TI - A malaria vaccine that elicits in humans antibodies able to kill Plasmodium falciparum. AB - BACKGROUND: Plasmodium falciparum merozoite surface protein 3 is a malaria vaccine candidate that was identified, characterised, and developed based on a unique immuno-clinical approach. The vaccine construct was derived from regions fully conserved among various strains and containing B cell epitopes targeted by human antibodies (from malaria-immune adults) that are able to mediate a monocyte dependent parasite killing effect. The corresponding long synthetic peptide was administered to 36 volunteers, with either alum or Montanide ISA720 as adjuvant. METHODS AND FINDINGS: Both formulations induced cellular and humoral immune responses. With alum, the responses lasted up to 12 mo. The vaccine-induced antibodies were predominantly of cytophilic classes, i.e., able to cooperate with effector cells. In vitro, the antibodies induced an inhibition of the P. falciparum erythrocytic growth in a monocyte-dependent manner, which was in most instances as high as or greater than that induced by natural antibodies from immune African adults. In vivo transfer of the volunteers' sera into P. falciparum-infected humanized SCID mice profoundly reduced or abrogated parasitaemia. These inhibitory effects were related to the antibody reactivity with the parasite native protein, which was seen in 60% of the volunteers, and remained in samples taken 12 mo postimmunisation. CONCLUSION: This is the first malaria vaccine clinical trial to clearly demonstrate antiparasitic activity by vaccine-induced antibodies by both in vitro and in vivo methods. The results, showing the induction of long-lasting antibodies directed to a fully conserved polypeptide, also challenge current concepts about malaria vaccines, such as unavoidable polymorphism, low antigenicity, and poor induction of immune memory. PMID- 16262452 TI - Evidence-based assessment of anxiety disorders in adults. AB - This article discusses issues related to the development and dissemination of evidence-based assessment strategies for anxiety disorders and associated problems. It begins with a review of the criteria that should be considered when determining whether particular assessment procedures are evidence-based. These include such factors as reliability, validity, cost-effectiveness, consumer acceptability, utility across different populations, and ease of dissemination. The importance of considering the purpose of the assessment process when deciding whether a procedure is evidence-based is emphasized. Next, the major assessment domains that are particularly important in the area of anxiety disorders (e.g., triggers for anxiety symptoms, avoidance behaviors) are reviewed. Finally, some potential obstacles to the dissemination of evidence-based assessment procedures are discussed along with suggestions for managing these obstacles. PMID- 16262453 TI - Evidence-based assessment of depression in adults. AB - From diverse perspectives, there is little doubt that depressive symptoms cohere to form a valid and distinct syndrome. Research indicates that an evidence-based assessment of depression would include (a) measures with adequate psychometric properties; (b) adequate coverage of symptoms; (c) adequate coverage of depressed mood, anhedonia, and suicidality; (d) an approach to suicidality that distinguishes between resolved plans and preparations and desire and ideation; (e) assessment of the atypical, seasonal, and melancholic subtypes; (f) parameters of course and chronicity; and (g) comorbidity and bipolarity. These complexities need to be accounted for when certain assessment approaches are preferred, and when ambiguity exists regarding the categorical versus dimensional nature of depression, and whether and when clinician ratings outperform self report. The authors conclude that no one extant procedure is ideal and suggest that the combination of certain interviews and self-report scales represents the state of the art for evidence-based assessment of depression. PMID- 16262454 TI - Evidence-based assessment of personality disorders. AB - The purpose of this article is to provide a foundation for the development of evidence-based guidelines for the assessment of personality disorders, focusing in particular on integrated assessment strategies. The general strategy recommended herein is to first administer a self-report inventory to alert oneself to the potential presence of particular maladaptive personality traits followed by a semistructured interview to verify their presence. This strategy is guided by the existing research that suggests particular strengths of self-report inventories and semistructured interviews relative to unstructured clinical interviews. However, the authors also consider research that suggests that further improvements to the existing instruments can be made. The authors emphasize, in particular, a consideration of age of onset, distortions in self perception and presentation, gender bias, culture and ethnicity, and personality change. PMID- 16262455 TI - Evidence-based approaches to assessing couple distress. AB - This article describes a conceptual framework for couple-based assessment strategies grounded in empirical findings linking couple distress to a broad range of both individual and relationship characteristics. These characteristics can contribute to, exacerbate, or result from relationship problems. On the basis of these findings, the authors articulate specific targets of clinical inquiry reflecting relationship behaviors, cognitions, and affect as well as features of individual distress. Guided by this framework, empirically supported assessment strategies and techniques emphasizing relationship functioning across diverse methods are proposed, including the clinical interview, analog behavioral observation, and both self- and other-report measures. Discussion concludes with specific recommendations regarding clinical assessment of couple distress and directions for further research. PMID- 16262456 TI - What's new about evidence-based assessment? AB - A clear consensus has emerged around the world concerning the desirability and even the urgency of basing health care delivery systems on evidence. Among behavioral health care providers such as psychologists, evidence-based practice (EBP) has been focused largely on interventions. Psychologists have long emphasized a scientifically based psychometric approach to the development of assessment procedures. Nevertheless, the era of evidence-based assessment highlights 2 somewhat different issues. First, sophisticated assessment is closely integrated with our emerging conceptions of psychopathology, rather than standing separate from these conceptions. Second, broad-based ongoing outcomes assessment systems are increasingly required for EBP on the part of governments and health care policymakers. This article summarizes these developments and looks to the future. PMID- 16262457 TI - Theory and utility--key themes in evidence-based assessment: comment on the special section. AB - This article focuses on two key themes in the four featured reviews on evidence based assessment. The first theme is the essential role of theory in psychological assessment. An overview of this complex, multilayered role is presented. The second theme is the need for a common metric with which to gauge the utility of specific psychological tests and measures for specific purposes. A metric from information theory is recommended. The implications of these themes for the four reviews and for the future of psychological assessment in general are discussed. PMID- 16262458 TI - Meta-analysis of the brief psychiatric rating scale factor structure. AB - A meta-analysis (N=17,620; k=26) of factor analyses of the Brief Psychiatric Rating Scale (BPRS) was conducted. Analysis of the 12 items from Overall et al.'s (J. E. Overall, L. E. Hollister, & P. Pichot, 1974) 4 subscales found support for his 4 subscales. Analysis of all 18 BPRS items found 4 components similar to those of Overall et al. In a 5-component solution, a 5th activation component emerged but was best supported among samples of schizophrenic patients. The first 4 components appear to form the core of the BPRS factor structure. Results of the meta-analysis suggest 5 subscales (with items in parentheses): Affect (anxiety, guilt, depression, somatic); Positive Symptoms (thought content, conceptual disorganization, hallucinatory behavior, grandiosity); Negative Symptoms (blunted affect, emotional withdrawal, motor retardation); Resistance (hostility, uncooperativeness, suspiciousness); and Activation (excitement, tension, mannerisms-posturing). PMID- 16262459 TI - Norming clinical questionnaires with multiple regression: the Pain Cognition List. AB - Questionnaires for measuring patients' feelings or beliefs are commonly used in clinical settings for diagnostic purposes, clinical decision making, or treatment evaluation. Raw scores of a patient can be evaluated by comparing them with norms based on a reference population. Using the Pain Cognition List (PCL-2003) as an example, this article shows how clinical questionnaires can be normed with multiple regression of raw scores on demographic and other patient variables. Compared with traditional norm tables for subgroups based on age or gender, this approach offers 2 advantages. First, multiple regression allows determination of which patient variables are relevant to the norming and which are not (validity). Second, by using information from the entire sample, multiple regression leads to continuous and more stable norms for any subgroup defined in terms of prognostic variables (reliability). PMID- 16262460 TI - Psychometric evaluation of the restructured clinical scales of the MMPI-2. AB - Conceptual overlap and heterogeneity have long been noted as weaknesses of the Minnesota Multiphasic Personality Inventory's clinical scales. Restructured clinical (RC) scales recently were developed to address these concerns (A. Tellegen et al., 2003). The authors evaluated the psychometric properties of the RC scales in psychology clinic clients (N=285) and military veterans (N=567). The RC scales were as internally consistent as the clinical scales and correlated strongly with their original counterparts (except for RC3/Hysteria). They also were less intercorrelated, produced conceptually clearer relations with measures of personality and psychopathology, and yielded somewhat greater incremental utility than the clinical scales. Thus, the RC scales demonstrated several psychometric strengths while utilizing 60% fewer items, but the 2 sets of scales cannot be used interchangeably. Interpretive considerations are discussed. PMID- 16262461 TI - An evaluation of evaluative personality terms: a comparison of the big seven and five-factor model in predicting psychopathology. AB - Two personality models are compared regarding their relationship with personality disorder (PD) symptom counts and with lifetime Axis I diagnoses. These models share 5 similar domains, and the Big 7 model also includes 2 domains assessing self-evaluation: positive and negative valence. The Big 7 model accounted for more variance in PDs than the 5-factor model, primarily because of the association of negative valence with most PDs. Although low-positive valence was associated with most Axis I diagnoses, the 5-factor model generally accounted for more variance in Axis I diagnoses than the Big 7 model. Some predicted associations between self-evaluation and psychopathology were not found, and unanticipated associations emerged. These findings are discussed regarding the utility of evaluative terms in clinical assessment. PMID- 16262462 TI - Latent structure of anxiety: taxometric exploration. AB - Taxometrics is a statistical tool that can be used to discern categories from continua. Taxometric analyses (MAXCOV and MAXEIG) were conducted in a large nonclinical sample (N=1,215) to determine whether extreme anxiety forms a distinct psychopathological category, an anxiety taxon. Anxiety was operationalized with self-report measures of subjective anxiety, anxious cognitive style, physiological arousal, and anxiety-related impairment. Procedures consistently identified a taxon with a prevalence of approximately 11%. Examination of the taxon's convergent and discriminant validity revealed that it reflects general distress rather than physiological arousal. Taxon membership showed some evidence of incremental validity. PMID- 16262463 TI - Recipient-provider agreement on enacted support, perceived support, and provider personality. AB - This study examined agreement between recipients and providers about social support and personality. One hundred daughter caregivers of a parent with Alzheimer's disease and each caregiver's most important support provider independently reported supportive behaviors provided to caregivers, the perceived supportiveness of the provider, and providers' personality traits. For all indices, agreement was higher for enacted support than for perceived support and personality, which were similar to each other for some, but not all, indices of agreement. These findings support the validity of measures of enacted and perceived support. PMID- 16262464 TI - Socially desirable responding and the factorial stability of the NEO PI-R. AB - The goal of the present investigation is to compare the factor structure of the revised NEO Personality Inventory (NEO PI-R; P. T. Costa & R. R. McCrae, 1992) in samples of respondents differentially motivated to respond in a socially desirable manner. In the French sample, the authors compared the NEO PI-R structure of job applicants (N=12,631) subgrouped by relative degree of socially desirable responding with that of a normative sample (N=801). In the Belgian sample, the authors compared the NEO PI-R structure in naturally occurring groups, including job applicants (N=3,028), individuals receiving career counseling (N=221), and a normative sample (N=549). Results indicate that the NEO PI-R factor structure remained stable across all samples. PMID- 16262466 TI - The dynamics of perceptual learning: an incremental reweighting model. AB - The mechanisms of perceptual learning are analyzed theoretically, probed in an orientation-discrimination experiment involving a novel nonstationary context manipulation, and instantiated in a detailed computational model. Two hypotheses are examined: modification of early cortical representations versus task-specific selective reweighting. Representation modification seems neither functionally necessary nor implied by the available psychophysical and physiological evidence. Computer simulations and mathematical analyses demonstrate the functional and empirical adequacy of selective reweighting as a perceptual learning mechanism. The stimulus images are processed by standard orientation- and frequency-tuned representational units, divisively normalized. Learning occurs only in the "read out" connections to a decision unit; the stimulus representations never change. An incremental Hebbian rule tracks the task-dependent predictive value of each unit, thereby improving the signal-to-noise ratio of their weighted combination. Each abrupt change in the environmental statistics induces a switch cost in the learning curves as the system temporarily works with suboptimal weights. PMID- 16262467 TI - The cultural mind: environmental decision making and cultural modeling within and across populations. AB - This article describes cross-cultural research on the relation between how people conceptualize nature and how they act in it. Mental models of nature differ dramatically among populations living in the same area and engaged in similar activities. This has novel implications for environmental decision making and management, including common problems. The research offers a distinct perspective on cultural modeling and a unified approach to studies of culture and cognition. The authors argue that cultural transmission and formation consist primarily not in shared rules or norms but in complex distributions of causally connected representations across minds interacting with the environment. The cultural stability and diversity of these representations often derive from rich, biologically prepared mental mechanisms that limit variation to readily transmissible psychological forms. This framework addresses several methodological issues, such as limitations on conceiving culture to be a well defined system, bounded entity, independent variable, or an internalized component of minds. PMID- 16262468 TI - SWIFT: a dynamical model of saccade generation during reading. AB - Mathematical models have become an important tool for understanding the control of eye movements during reading. Main goals of the development of the SWIFT model (R. Engbert, A. Longtin, & R. Kliegl, 2002) were to investigate the possibility of spatially distributed processing and to implement a general mechanism for all types of eye movements observed in reading experiments. The authors present an advanced version of SWIFT that integrates properties of the oculomotor system and effects of word recognition to explain many of the experimental phenomena faced in reading research. They propose new procedures for the estimation of model parameters and for the test of the model's performance. They also present a mathematical analysis of the dynamics of the SWIFT model. Finally, within this framework, they present an analysis of the transition from parallel to serial processing. PMID- 16262469 TI - An anatomically constrained, stochastic model of eye movement control in reading. AB - This article presents SERIF, a new model of eye movement control in reading that integrates an established stochastic model of saccade latencies (LATER; R. H. S. Carpenter, 1981) with a fundamental anatomical constraint on reading: the vertically split fovea and the initial projection of information in either visual field to the contralateral hemisphere. The novel features of the model are its simulation of saccade latencies as a race between two stochastic rise-to threshold LATER units and its probabilistic selection of the target for the next saccade. The model generates simulated eye movement behavior that exhibits important characteristics of actual eye movements made during reading; specifically, simulations produce realistic saccade target distributions and replicate a number of critical reading phenomena, including the effects of word frequency on fixation durations, the inverted optimal viewing position effect, the trade-off between first and second fixation durations of refixated words, and the dependence of parafoveal preview benefit on eccentricity. PMID- 16262470 TI - A dynamic, stochastic, computational model of preference reversal phenomena. AB - Preference orderings among a set of options may depend on the elicitation method (e.g., choice or pricing); these preference reversals challenge traditional decision theories. Previous attempts to explain these reversals have relied on allowing utility of the options to change across elicitation methods by changing the decision weights, the attribute values, or the combination of this information--still, no theory has successfully accounted for all the phenomena. In this article, the authors present a new computational model that accounts for the empirical trends without changing decision weights, values, or combination rules. Rather, the current model specifies a dynamic evaluation and response process that correctly predicts preference orderings across 6 elicitation methods, retains stable evaluations across methods, and makes novel predictions regarding response distributions and response times. PMID- 16262471 TI - Modeling behavior in a clinically diagnostic sequential risk-taking task. AB - This article models the cognitive processes underlying learning and sequential choice in a risk-taking task for the purposes of understanding how they occur in this moderately complex environment and how behavior in it relates to self reported real-world risk taking. The best stochastic model assumes that participants incorrectly treat outcome probabilities as stationary, update probabilities in a Bayesian fashion, evaluate choice policies prior to rather than during responding, and maintain constant response sensitivity. The model parameter associated with subjective value of gains correlates well with external risk taking. Both the overall approach, which can be expanded as the basic paradigm is varied, and the specific results provide direction for theories of risky choice and for understanding risk taking as a public health problem. PMID- 16262472 TI - Absolute identification by relative judgment. AB - In unidimensional absolute identification tasks, participants identify stimuli that vary along a single dimension. Performance is surprisingly poor compared with discrimination of the same stimuli. Existing models assume that identification is achieved using long-term representations of absolute magnitudes. The authors propose an alternative relative judgment model (RJM) in which the elemental perceptual units are representations of the differences between current and previous stimuli. These differences are used, together with the previous feedback, to respond. Without using long-term representations of absolute magnitudes, the RJM accounts for (a) information transmission limits, (b) bowed serial position effects, and (c) sequential effects, where responses are biased toward immediately preceding stimuli but away from more distant stimuli (assimilation and contrast). PMID- 16262473 TI - On adaptation, maximization, and reinforcement learning among cognitive strategies. AB - Analysis of binary choice behavior in iterated tasks with immediate feedback reveals robust deviations from maximization that can be described as indications of 3 effects: (a) a payoff variability effect, in which high payoff variability seems to move choice behavior toward random choice; (b) underweighting of rare events, in which alternatives that yield the best payoffs most of the time are attractive even when they are associated with a lower expected return; and (c) loss aversion, in which alternatives that minimize the probability of losses can be more attractive than those that maximize expected payoffs. The results are closer to probability matching than to maximization. Best approximation is provided with a model of reinforcement learning among cognitive strategies (RELACS). This model captures the 3 deviations, the learning curves, and the effect of information on uncertainty avoidance. It outperforms other models in fitting the data and in predicting behavior in other experiments. PMID- 16262474 TI - A model for stochastic drift in memory strength to account for judgments of learning. AB - Previous research has shown that judgments of learning (JOLs) made immediately after encoding have a low correlation with actual cued-recall performance, whereas the correlation is high for delayed judgments. In this article, the authors propose a formal theory describing the stochastic drift of memory strength over the retention interval to account for the delayed-JOL effect. This is done by first decomposing the aggregated memory strength into exponential functions with slow and fast memory traces. The mean aggregated memory strength shows power-function forgetting curves. The drift of the memory strength is large for immediate JOLs (causing a low predictability) and weak for delayed JOLs (causing a high predictability). Consistent with empirical data, the model makes a novel prediction of JOL asymmetry, or that immediate weak JOLs are more predictive of future performance than are immediate strong JOLs. The JOL distributions for immediate and delayed JOLs are also accounted for. PMID- 16262475 TI - Why most people disapprove of me: experience sampling in impression formation. AB - Individuals are typically more likely to continue to interact with people if they have a positive impression of them. This article shows how this sequential sampling feature of impression formation can explain several biases in impression formation. The underlying mechanism is the sample bias generated when the probability of interaction depends on current impressions. Because negative experiences decrease the probability of interaction, negative initial impressions are more stable than positive impressions. Negative initial impressions, however, are more likely to change for individuals who are frequently exposed to others. As a result, systematic differences in interaction patterns, due to social similarity or proximity, will produce systematic differences in impressions. This mechanism suggests an alternative explanation of several regularities in impression formation, including a negativity bias in impressions of outgroup members, systematic differences in performance evaluations, and more positive evaluations of proximate others. PMID- 16262476 TI - Finding useful questions: on Bayesian diagnosticity, probability, impact, and information gain. AB - Several norms for how people should assess a question's usefulness have been proposed, notably Bayesian diagnosticity, information gain (mutual information), Kullback-Liebler distance, probability gain (error minimization), and impact (absolute change). Several probabilistic models of previous experiments on categorization, covariation assessment, medical diagnosis, and the selection task are shown to not discriminate among these norms as descriptive models of human intuitions and behavior. Computational optimization found situations in which information gain, probability gain, and impact strongly contradict Bayesian diagnosticity. In these situations, diagnosticity's claims are normatively inferior. Results of a new experiment strongly contradict the predictions of Bayesian diagnosticity. Normative theoretical concerns also argue against use of diagnosticity. It is concluded that Bayesian diagnosticity is normatively flawed and empirically unjustified. PMID- 16262477 TI - The function of phenomenal states: supramodular interaction theory. AB - Discovering the function of phenomenal states remains a formidable scientific challenge. Research on consciously penetrable conflicts (e.g., "pain-for-gain" scenarios) and impenetrable conflicts (as in the pupillary reflex, ventriloquism, and the McGurk effect [H. McGurk & J. MacDonald, 1976]) reveals that these states integrate diverse kinds of information to yield adaptive action. Supramodular interaction theory proposes that phenomenal states play an essential role in permitting interactions among supramodular response systems--agentic, independent, multimodal, information-processing structures defined by their concerns (e.g., instrumental action vs. certain bodily needs). Unlike unconscious processes (e.g., pupillary reflex), these processes may conflict with skeletal muscle plans, as described by the principle of parallel responses into skeletal muscle (PRISM). Without phenomenal states, these systems would be encapsulated and incapable of collectively influencing skeletomotor action. PMID- 16262478 TI - Meaning through syntax is insufficient to explain comprehension of sentences with reduced relative clauses: comment on McKoon and Ratcliff (2003). AB - The authors argue that the meaning through syntax (MTS) model proposed by G. McKoon and R. Ratcliff fails to account for the comprehension of sentences with reduced relative clauses. First, the theory's core assumptions regarding verb based event representations and how they link to constructions are incompatible with well-established analyses from the lexical semantics literature. Second, the MTS theory provides neither a principled nor a consistent account for why some reduced relatives are hard whereas others are easy. Finally, McKoon and Ratcliff's critique of constraint-based models is flawed in that sometimes they tested a nonexistent theory and sometimes they provided evidence for the constraint-based models against which they were arguing. PMID- 16262481 TI - Suppositions, extensionality, and conditionals: a critique of the mental model theory of Johnson-Laird And Byrne (2002). AB - P. N. Johnson-Laird and R. M. J. Byrne proposed an influential theory of conditionals in which mental models represent logical possibilities and inferences are drawn from the extensions of possibilities that are used to represent conditionals. In this article, the authors argue that the extensional semantics underlying this theory is equivalent to that of the material, truth functional conditional, at least for what they term "basic" conditionals, concerning arbitrary problem content. On the basis of both logical argument and psychological evidence, the authors propose that this approach is fundamentally mistaken and that conditionals must be viewed within a suppositional theory based on what philosophical logicians call the Ramsey test. The Johnson-Laird and Byrne theory is critically examined with respect to its account of basic conditionals, nonbasic conditionals, and counterfactuals. PMID- 16262482 TI - Orienting to eye gaze and face processing. AB - The author conducted 7 experiments to examine possible interactions between orienting to eye gaze and specific forms of face processing. Participants classified a letter following either an upright or inverted face with averted, uninformative eye gaze. Eye gaze orienting effects were recorded for upright and inverted faces, irrespective of whether the faces were simple, schematic faces or more realistic faces. In contrast, inversion affected orienting to targets appearing along the vertical axis. Switching the contrast between the iris and sclera reversed orienting to eye gaze. Lifting the eyelid to expose more of the iris-sclera contrast led to a potentiation of orienting to eye gaze. Raising the eyebrow alone without the eyelid did not affect orienting. The findings suggest that local perceptual information is critical for orienting to eye gaze and that the effect can occur with a degree of independence from certain types of face processing. PMID- 16262483 TI - Prioritization of new objects in real-world scenes: evidence from eye movements. AB - The authors examined the prioritization of abruptly appearing objects in real world scenes by measuring the eyes' propensity to be directed to the new object. New objects were fixated more often than chance whether they appeared during fixations (transient onsets) or saccades (nontransient onsets). However, onsets that appeared during fixations were fixated sooner and more often than those coincident with saccades. Prioritization of onsets during saccades, but not fixations, were affected by manipulations of memory: Reducing scene viewing time prior to the onset eliminated prioritization, whereas prior study of the scenes increased prioritization. Transient objects draw attention quickly and do not depend on memory, but without a transient signal, new objects are prioritized over several saccades as memory is used to explicitly identify the change. These effects were not modulated by observers' expectations concerning the appearance of new objects, suggesting the prioritization of a transient is automatic and that memory-guided prioritization is implicit. PMID- 16262484 TI - Overloading temporal memory. AB - This study tested the hypothesis that memory is a major source of variance in temporal processing. Participants categorized intervals as short or long. The number of base durations and interval types mixed within blocks of trials varied from 1 session to another. Results revealed that mixing 2 base durations within blocks increased categorization errors, but mixing 2 marker types did not. Results are attributed to the involvement of more than 1 memory representation, which is argued to show the critical role of memory in temporal processing. Because mixing modalities has no such effect, it was argued that modalities share a common representation in memory. Finally, there was no difference in the perceived duration of auditory and visually marked intervals, which is inconsistent with most reports on this effect. PMID- 16262485 TI - Tool use affects perceived distance, but only when you intend to use it. AB - Recent research demonstrates neurologic and behavioral differences in people's responses to the space that is within and beyond reach. The present studies demonstrated a perceptual difference as well. Reachability was manipulated by having participants reach with and without a tool. Across 2 conditions, in which participants either held a tool or not, targets were presented at the same distances. Perceived distances to targets within reach holding the tool were compressed compared with targets that were beyond reach without it. These results suggest that reachability serves as a metric for perception. The 3rd experiment found that reachability only influenced perceived distance when the perceiver intended to reach. These experiments suggest that the authors perceive the environment in terms of our intentions and abilities to act within it. PMID- 16262486 TI - Associative learning improves visual working memory performance. AB - The ability to remember visual stimuli over a short delay period is limited by the small capacity of visual working memory (VWM). Here the authors investigate the role of learning in enhancing VWM. Participants saw 2 spatial arrays separated by a 1-s interval. The 2 arrays were identical except for 1 location. Participants had to detect the difference. Unknown to the participants, some spatial arrays would repeat once every dozen trials or so for up to 32 repetitions. Spatial VWM performance increased significantly when the same location changed across display repetitions, but not at all when different locations changed from one display repetition to another. The authors suggest that a major role of learning in VWM is to mediate which information gets retained, rather than to directly increase VWM capacity. PMID- 16262487 TI - The role of visual and nonvisual information in the control of locomotion. AB - During locomotion, retinal flow, gaze angle, and vestibular information can contribute to one's perception of self-motion. Their respective roles were investigated during active steering: Retinal flow and gaze angle were biased by altering the visual information during computer-simulated locomotion, and vestibular information was controlled through use of a motorized chair that rotated the participant around his or her vertical axis. Chair rotation was made appropriate for the steering response of the participant or made inappropriate by rotating a proportion of the veridical amount. Large steering errors resulted from selective manipulation of retinal flow and gaze angle, and the pattern of errors provided strong evidence for an additive model of combination. Vestibular information had little or no effect on steering performance, suggesting that vestibular signals are not integrated with visual information for the control of steering at these speeds. PMID- 16262488 TI - The perception of second language sounds in early bilinguals: new evidence from an implicit measure. AB - Previous studies have suggested that nonnative (L2) linguistic sounds are accommodated to native language (L1) phonemic categories. However, this conclusion may be compromised by the use of explicit discrimination tests. The present study provides an implicit measure of L2 phoneme discrimination in early bilinguals (Catalan and Spanish). Participants classified the 1st syllable of disyllabic stimuli embedded in lists where the 2nd, task-irrelevant, syllable could contain a Catalan contrastive variation (/epsilon/-/e/) or no variation. Catalan dominants responded more slowly in lists where the 2nd syllable could vary from trial to trial, suggesting an indirect effect of the /epsilon/-/e/ discrimination. Spanish dominants did not suffer this interference, performing indistinguishably from Spanish monolinguals. The present findings provide implicit evidence that even proficient bilinguals categorize L2 sounds according to their L1 representations. PMID- 16262489 TI - Identification and bisection of temporal durations and tone frequencies: common models for temporal and nontemporal stimuli. AB - Two experiments examined identification and bisection of tones varying in temporal duration (Experiment 1) or frequency (Experiment 2). Absolute identification of both durations and frequencies was influenced by prior stimuli and by stimulus distribution. Stimulus distribution influenced bisection for both stimulus types consistently, with more positively skewed distributions producing lower bisection points. The effect of distribution was greater when the ratio of the largest to smallest stimulus magnitude was greater. A simple mathematical model, temporal range frequency theory, was applied. It is concluded that (a) similar principles describe identification of temporal durations and other stimulus dimensions and (b) temporal bisection point shifts can be understood in terms of psychophysical principles independently developed in nontemporal domains, such as A. Parducci's (1965) range frequency theory. PMID- 16262490 TI - Rhythmic masking release: effects of asynchrony, temporal overlap, harmonic relations, and source separation on cross-spectral grouping. AB - The rhythm created by spacing a series of brief tones in a regular pattern can be disguised by interleaving identical distractors at irregular intervals. The disguised rhythm can be unmasked if the distractors are allocated to a separate stream from the rhythm by integration with temporally overlapping captors. Listeners identified which of 2 rhythms was presented, and the accuracy and rated clarity of their judgment was used to estimate the fusion of the distractors and captors. The extent of fusion depended primarily on onset asynchrony and degree of temporal overlap. Harmonic relations had some influence, but only an extreme difference in spatial location was effective (dichotic presentation). Both preattentive and attentionally driven processes governed performance. PMID- 16262491 TI - Eye movements and word skipping during reading revisited. AB - The authors examined word skipping in reading in 2 experiments. In Experiment 1, skipping rates were higher for a preview of a predictable word than for a visually similar nonword, indicating there is full recognition in parafoveal vision. In Experiment 2, foveal load was manipulated by varying the frequency of the word preceding either a 3-letter target word or a misspelled preview. There was again a higher skipping rate for a correct preview and a lower skipping rate when there was a high foveal load, but there was no interaction, and the pattern of effects in fixation times was the same as in the skipping data. Experiment 2 also showed significant skipping of nonwords similar to the target word, indicating skipping based on partial information. PMID- 16262492 TI - Especial skills: their emergence with massive amounts of practice. AB - Differing viewpoints concerning the specificity and generality of motor skill representations in memory were compared by contrasting versions of a skill having either extensive or minimal specific practice. In Experiments 1 and 2, skilled basketball players more accurately performed set shots at the foul line than would be predicted on the basis of the performance at the nearby locations, suggesting considerable specificity at this distance. This effect was replicated even when the lines on the court were obscured (in Experiment 2). However, the effect was absent when jump shots were executed in Experiment 3. The authors argue that massive levels of practice at 1 particular member of a class of actions produce specific effects that allow this skill to stand out from the other members of the class, giving it the status of an especial skill. Various theoretical views are proposed to account for the development of these skills. PMID- 16262494 TI - How perception impacts on drawings. AB - In 3 experiments the authors investigate how errors in perception produce errors in drawings. In Experiment 1, when Shepard stimuli were shown as a pair of tables, participants made severe errors in trying to adjust 1 part of the stimulus to match the other. When the table legs were removed, revealing a pair of parallelograms with minimal perspective cues, the illusion was weaker. The authors predicted that participants would err when drawing the table but not the parallelogram stimuli. The results of Experiment 2 support the prediction and establish a direct link between degree of perceptual distortion of the table stimuli and the severity of error in drawing. When drawing only the right-hand part of the figure, participants also erred to a greater degree in drawing the table than the parallelogram (Experiment 3). Collectively, the results suggest that perceptual distortion is linked with errors in drawing the table stimuli. PMID- 16262493 TI - Time course of linguistic information extraction from consecutive words during eye fixations in reading. AB - Sequential attention shift models of reading predict that an attended (typically fixated) word must be recognized before useful linguistic information can be obtained from the following (parafoveal) word. These models also predict that linguistic information is obtained from a parafoveal word immediately prior to a saccade toward it. To test these assumptions, sentences were constructed with a critical pretarget-target word sequence, and the temporal availability of the (parafoveal) target preview was manipulated while the pretarget word was fixated. Target viewing effects, examined as a function of prior target visibility, revealed that extraction of linguistic target information began 70-140 ms after the onset of pretarget viewing. Critically, acquisition of useful linguistic information from a target was not confined to the ending period of pretarget viewing. These results favor theoretical conceptions in which there is some temporal overlap in the linguistic processing of a fixated and parafoveally visible word during reading. PMID- 16262495 TI - Different strategies for using motion-in-depth information in catching. AB - Previous studies on ball catching have had the limitation that the catcher was restricted to lateral hand movements. The authors investigated catching behavior in the more natural situation in which hand movements were unconstrained. Movements of the hand were tracked as participants tried to "catch" an approaching ball simulated with changing size and/or changing disparity. Participants used 1 of 2 distinct interception strategies: (a) a "cutting-off" strategy where time to passage (TTP) information was used to guide movements of the hand in depth such that the ball was caught farther in front of the face when the ball was approaching more slowly and (b) a "waiting" strategy where the hand was moved along a frontoparallel plane that was constant across ball trajectories and speeds. Cue dissociation and selective adaptation manipulations demonstrated that the catcher's estimates of TTP and crossing distance were based on a combination of binocular and monocular information. PMID- 16262496 TI - Naive optics: acting on mirror reflections. AB - It is known that naive observers have striking misconceptions about mirror reflections. In 5 experiments, this article systematically extends the findings to graphic stimuli, to interactive visual tasks, and finally to tasks involving real mirrors. The results show that the perceptual knowledge of nonexpert adults is far superior to their conceptual knowledge. Whereas conceptual errors include the assumption of left-right reversals in mirror images and often blatant extensions of the boundary of mirror space, the perceptual context prevents most such errors. However, a consistent bias to misjudge objects in mirrors too far to the outside is demonstrable in all cases including tasks with real mirrors. The authors present a 2-stage hypothesis consisting of an implicit bias of judging the mirror surface to be turned toward the observer's line of sight followed by a normalization that becomes explicit. PMID- 16262497 TI - Attentional capture by an unannounced color singleton depends on expectation discrepancy. AB - Eight experiments examined the conditions under which a color singleton that is presented for the 1st time without prior announcement captures attention. The main hypothesis is that an unannounced singleton captures attention to the extent that it deviates from expectations. This hypothesis was tested within a visual search paradigm in which set-size effects were used to infer attentional capture. The results showed that attentional capture by an unannounced color singleton was due to a mismatch with expectations concerning the color of the object and not due to its being a singleton. Thus, the results imply that theories of attention have to consider expectation discrepancy as a determinant of attention shifts. PMID- 16262498 TI - Attending to eye movements and retinal eccentricity: evidence for the activity distribution model of attention reconsidered. AB - When testing between spotlight and activity distribution models of visual attention, D. LaBerge, R. L. Carlson, J. K. Williams, and B. G. Bunney (1997) used an experimental paradigm in which targets are embedded in 3 brief displays. This paradigm, however, may be confounded by retinal eccentricity effects and saccadic eye movements. When the retinal eccentricities of the targets are equated and eye position is monitored, the pattern of results reported by LaBerge et al., which supported the activity distribution model, is not found. This result underscores the importance of considering targets' eccentricity and people's inclination to make saccadic eye movements in certain types of visual cognition tasks. ((c) 2005 APA, all rights reserved). PMID- 16262499 TI - How much attention does an event file need? AB - Attentional requirements for the spontaneous integration of stimulus and response features were analyzed. In line with previous findings, carrying out a prepared response to the onset of a stimulus created bindings between the response and the features of that stimulus, thereby impairing subsequent performance on mismatching stimulus-response combinations. Findings demonstrate that a stimulus gets bound to a response even if its presence is neither necessary nor useful for the task at hand, it follows rather than precedes the response in time, it competes with a task-relevant stimulus, and if the response is suppressed--but only if the stimulus appears close to the response's eventual execution or abandonment. A multiple-integration model is suggested that assumes that the integration of stimulus features in perception and of response features in action planning are local processes that are independent of stimulus-response integration, which presumably is triggered by the success of the perception action episode. PMID- 16262500 TI - Characterizing the motor execution stage of speech production: consonantal effects on delayed naming latency and onset duration. AB - The research described in this article had 2 aims: to permit greater precision in the conduct of naming experiments and to contribute to a characterization of the motor execution stage of speech production. The authors report an exhaustive inventory of consonantal and postconsonantal influences on delayed naming latency and onset acoustic duration, derived from a hand-labeled corpus of single syllable consonant-vowel utterances. Five talkers produced 6 repetitions each of a set of 168 prepared monosyllables, a set that comprised each of the consonantal onsets of English in 3 vowel contexts. Strong and significant effects associated with phonetic characteristics of initial and noninitial phonemes were observed on both delayed naming latency and onset acoustic duration. Results are discussed in terms of the biomechanical properties of the articulatory system that may give rise to these effects and in terms of their methodological implications for naming experiments. PMID- 16262501 TI - What learning to see arbitrary motion tells us about biological motion perception. AB - In separate studies, observers viewed upright biological motion, inverted biological motion, or arbitrary motion created from systematically randomizing the positions of point-light dots. Results showed that observers (a) could learn to detect the presence of arbitrary motion, (b) could not learn to discriminate the coherence of arbitrary motion, although they could do so for upright biological motion, (c) could apply a detection strategy to learn to detect the presence of inverted biological motion nearly as well as they detected upright biological motion, and (d) performed better discriminating the coherence of upright biological motion compared with inverted biological motion. These results suggest that learning and form information play an important role in perceiving biological motion, although this role may only be apparent in tasks that require processing information from multiple parts of the motion display. PMID- 16262502 TI - The scaling of information to action in visually guided braking. AB - Braking to avoid a collision can be controlled by keeping the deceleration required to stop (i.e., ideal deceleration) in the "safe" region below maximum deceleration, but maximum deceleration is not optically specified and can vary as conditions change. When brake strength was manipulated between participants using a simulated braking task, the ratio of ideal to maximum deceleration at brake onset was invariant across groups, suggesting that calibration involves scaling information about ideal deceleration in intrinsic units of maximum deceleration. Evidence of rapid recalibration was found when brake strength was manipulated within participants, and the presence of external forces that affect brake dynamics resulted in biases in performance. Discussion focuses on the role of calibration, internal models, and affordance perception in visually guided action. PMID- 16262503 TI - Perceptual tests of an algorithm for musical key-finding. AB - Perceiving the tonality of a musical passage is a fundamental aspect of the experience of hearing music. Models for determining tonality have thus occupied a central place in music cognition research. Three experiments investigated 1 well known model of tonal determination: the Krumhansl-Schmuckler key-finding algorithm. In Experiment 1, listeners' percepts of tonality following short musical fragments derived from preludes by Bach and Chopin were compared with predictions of tonality produced by the algorithm; these predictions were very accurate for the Bach preludes but considerably less so for the Chopin preludes. Experiment 2 explored a subset of the Chopin preludes, finding that the algorithm could predict tonal percepts on a measure-by-measure basis. In Experiment 3, the algorithm predicted listeners' percepts of tonal movement throughout a complete Chopin prelude. These studies support the viability of the Krumhansl-Schmuckler key-finding algorithm as well as a model of listeners' tonal perceptions of musical passages. PMID- 16262504 TI - Naming two-digit arabic numerals: evidence from masked priming studies. AB - The authors investigated how 2-digit Arabic numerals are named by looking at the effects of masked primes on the naming latencies. Target numerals were named faster when prime and target shared a digit at the same position (e.g., the target 28 primed by 18 and 21). In contrast, naming latencies were slower when prime and target shared 1 or 2 digits at non-corresponding places (e.g., the target 28 primed by 82, 86, or 72). Subsequent experiments showed that these priming effects were situated at the level of the verbal production of the Arabic numerals. The data point to a non-semantically mediated route from visual input to verbal output in the naming of 2-digit Arabic numerals. PMID- 16262505 TI - Trauma and cumulative adversity in women of a disadvantaged social location. AB - This study expands conceptual and empirical perspectives on stress exposure by evaluating cumulative exposure in 79 drug-convicted women. Logistic regression determined that (a) posttraumatic stress disorder (PTSD) increased 40% with each trauma and (b) adding chronic stressors increased the predictability of PTSD. This study supports cumulative adversity models and the importance of social location. PMID- 16262506 TI - A preventive pilot project addressing multiethnic tensions in the wake of the Iraq war. AB - This article describes a school-based preventive pilot project for recent immigrant children, designed to decrease anxiety and intergroup tensions associated with the Iraq war. Results suggest that clinicians should address the multiplicity of meanings associated with international events when planning a prevention program in multiethnic schools to help children to cope with the increasingly common gap between the ways traumatic events covered by the media are understood at home and at school. PMID- 16262507 TI - The orphans of Eritrea: what are the choices? AB - The authors examined the emotional well-being, adaptability, and emotional distress of 10-13 year-old Eritrean war orphans cared for in 3 different social environments and 1 group of home-reared children using 2 scales of the Behavioral Assessment System for Children and informal interviews with the children. Orphans reunified with extended families had greater adaptive skills than institutional orphans but as many signs and symptoms of emotional distress as orphanage children. Group-home orphans had fewer signs and symptoms of emotional distress and greater adaptive skills than either reunified or institutional orphans, and they had fewer symptoms of emotional distress than home-reared children. However, placing orphans in small group homes was far more expensive than reunifying them with extended families. The public policy implications of the findings for the protection of unaccompanied children in impoverished developing countries are discussed. PMID- 16262508 TI - Children's experience of loss by parental migration in inner-city Jamaica. AB - Migratory separation, when parents migrate and leave their children behind, was investigated in a case-control sample of 9- to 10-year-olds living in inner-city communities in Kingston and St Andrew, Jamaica (N = 54). Data analyses using descriptive statistics and bivariate correlations showed that children's reactions to their parents' migration were directly related to poor school performance and psychological difficulties. Additionally, being currently exposed to violence in the home and/or community was significantly associated with high scores on a measure of grief intensity. "Protective" factors included having someone to talk to about the migration and living in a supportive family. Migratory separation needs detailed investigation like that devoted to other childhood family disruptions, such as parental divorce or death. PMID- 16262509 TI - The relationship between violence exposure and HIV sexual risk behavior: does gender matter? AB - This study examined the association between violence exposures and HIV risk behaviors among 409 adolescents and whether such associations are gendered. Results indicated that adolescents exposed to either childhood sexual abuse or family or community violence were almost 3 and 4 times more likely, respectively, than peers not exposed to such violence to report a higher number of HIV-related risk behaviors. In addition, boys exposed to family violence were almost 3 times more likely than girls to report multiple partners and use of drugs during sex. Findings suggest a complex relationship among violence, gender, and HIV risks among adolescents. PMID- 16262510 TI - "It's just not all teenage moms": diversity, support, and relationship in family services. AB - Social support is often used in family services, implicitly or explicitly, as a tool for enhancing family functioning. Using data from a focus group study, this article explores aspects of the relationships parents develop through participation in a family support program. Program structures and processes shaping relationships are also considered. Findings are then interpreted through a social capital theoretical lens, and implications for practice are discussed. PMID- 16262511 TI - The national lesbian family study: 4. Interviews with the 10-year-old children. AB - This 4th report from a longitudinal study of U.S. lesbian families presents data from 78 families in which the children were conceived by donor insemination. Results indicate that the prevalence of physical and sexual abuse in these children was lower than national norms. In social and psychological development, the children were comparable to children raised in heterosexual families. Children of unknown donors were indistinguishable from those with known donors in psychological adjustment. In total, 57% of the children were completely out to their peers, and 43% had experienced homophobia. The children demonstrated a sophisticated understanding of diversity and tolerance. PMID- 16262512 TI - Implementing trauma-informed alcohol and other drug and mental health services for women: lessons learned in a multisite demonstration project. AB - On the basis of the 9-site, Substance Abuse and Mental Health Services Administration-funded Women, Co-Occurring Disorders, and Violence Study, this article discusses recommendations for implementing trauma-informed mental health, substance abuse, and other support services. These guidelines for best practices represent the consensus of a diverse trauma work group that drew on both cross site and site-specific qualitative data. PMID- 16262513 TI - Beyond treatment effects: predicting emerging adult alcohol and marijuana use among substance-abusing delinquents. AB - Secondary analyses of a randomized clinical trial examined the effects of 4 putative risk factors and 2 protective factors in predicting drug use among 80 emerging adults treated 5 years earlier for delinquency and alcohol and/or marijuana use disorders. Frequency of marijuana use and the number of comorbid psychiatric disorders in adolescence predicted cannabis use in emerging adulthood. Increasing academic competence at high levels of social competence predicted less marijuana use. At emerging adulthood, greater use of alcohol and marijuana were associated with both criminality and psychopathology. PMID- 16262514 TI - Eating disorders in Asian populations: a critique of current approaches to the study of culture, ethnicity, and eating disorders. AB - There is increasing evidence that eating disorders are present among ethnically diverse populations, and researchers have suggested that investigations in this area may inform the field's understanding of how sociocultural factors are related to the development of eating disorders. Although it is generally accepted that sociocultural factors are key in eating disorder etiology, knowledge on how best to study these influences in diverse groups is still limited. In this article, the authors review how the research literature has explored relationships among culture, ethnicity, and eating disorders in Asian populations and critically examine strategies that have been used to investigate these issues across 1 ethnic/racial group. The methodological challenges encountered in these approaches are identified and considered in the provision of recommendations for future endeavors to improve the field's understanding of how culture is related to eating disorders. PMID- 16262515 TI - The relation between violence exposure and conduct problems among adolescents: a prospective study. AB - This study tested the hypothesis that exposure to community violence, intimate partner violence, and child maltreatment independently contribute to the prediction of conduct problems over a 2-year period. Participants were a subsample of youth ages 12 to 17 years (N = 423) from the Patterns of Care study, which drew a stratified random sample of high-risk youth receiving services from public service sectors. Exposure to community violence significantly predicted conduct disorder and externalizing problems 2 years later when potential confounds were controlled. Child maltreatment predicted conduct disorder but not externalizing symptoms. Exposure to intimate partner violence was not related to either outcome. Exposure to community violence contributed to the development of conduct disorder and externalizing symptoms, even when exposure to child maltreatment or intimate partner violence was controlled. Results are discussed in terms of implications for treatment and prevention of youth conduct problems. PMID- 16262516 TI - God forbid! Substance use among religious and non-religious youth. AB - Among a predominantly Mexican and Mexican American sample of preadolescents, religiosity protected against lifetime alcohol, cigarette, and marijuana use and recent alcohol and cigarette use when religious affiliation was controlled. When religiosity was controlled, however, adolescents with no religious affiliation and adolescents who were religiously affiliated reported similar substance use outcomes. Interaction effects demonstrated that the protective effect of greater religiosity operated more strongly in some religions than in others for selected outcomes. Overall, the impact of religiosity on reported drug use did not differ significantly for more and less acculturated Latino youth. PMID- 16262517 TI - Relations between parents' interactive style in dyadic and triadic play and toddlers' symbolic capacity. AB - Play has a major role in the evaluation and treatment of young children referred to mental health clinicians. The present study examined parental correlates of preschoolers' symbolic play during dyadic and triadic play interactions. Boys' play contained more aggressive themes, and girls' contained more nurturing themes. Mothers displayed more caring themes during play with both sons and daughters, and fathers displayed more repair and construction themes. Mothers' and fathers' facilitative- creative interaction style in dyadic play predicted the level of the child's symbolic play. Co-parenting style marked by cooperation and autonomy predicted symbolic play during a triadic family session. Child intelligence predicted symbolic play beyond the parent's style during triadic but not dyadic interactions. The findings have implications for early intervention directed at increasing symbolic play in young children. PMID- 16262518 TI - The parental empathy measure: a new approach to assessing child maltreatment risk. AB - A new operational definition of parental empathy and a new instrument, the Parental Empathy Measure (PEM), are introduced. With a sample of 103 parents (50 registered maltreating, 32 matched distressed, and 21 matched controls), the PEM demonstrated good internal consistency, very good interrater reliability, good construct validity, and very good concurrent validity. PMID- 16262519 TI - Dyadic characteristics of individual attributes: attachment, neuroticism, and their relation to marital quality and closeness. AB - The present article focuses on couple types based on 2 personality traits, attachment security and neuroticism, as they relate to 2 facets of the marital relationship--a global evaluation of relationship quality and dyadic closeness distance. The sample consisted of 248 married couples who completed measures of attachment anxiety and avoidance, neuroticism, and marital quality, as well as levels of closeness reported over 7 consecutive days. Cluster analyses yielded 3 types of dyadic attachment configurations (secure, fearful avoidant, and insecure mixed) and 4 types of dyadic neuroticism (low couple neuroticism, high couple neuroticism, wife neuroticism, and husband neuroticism). Significant differences were found among attachment and neuroticism dyadic types in marital quality. The findings are discussed in terms of the viability of dyadic types based on individual traits, implying that attachment security yields itself to dyadic conceptualization more than neuroticism. PMID- 16262521 TI - Role taking of youths in a family context: adolescents exposed to interparental violence. AB - This qualitative study examines the roles enacted by adolescents who lived with interparental violence and the impact of such roles on the formation of their identity. The study sample was composed of 21 Israeli-born Jewish youths ages 13 18 years. Data analysis revealed an emic perspective on the roles enacted by youths at various points in the development of interparental conflict. These roles were described and analyzed in relation to the temporal and spatial dimensions of violent situations and according to the type of involvement, the range of authority and responsibility assumed by the youths, and the extent of choice they exercised in role taking. The results are discussed in terms of current role theory, and some practical implications are suggested. PMID- 16262520 TI - Preserving family bonds: examining parent perspectives in the light of practice standards for out-of-home treatment. AB - Family participation is a core system of care value that is supported by previous research in medical, child welfare, and mental health settings. However, many parents with children receiving out-of-home mental health treatment experience restrictions on contact. This cross-sectional study examines the experiences of families (N = 102) regarding parent-child contact in relation to examples of national accreditation standards. Results of the national survey found that most respondents (79.4%) reported restrictions on contact, including limits based on behavioral contingencies (65.7%) and point and level systems (52.5%). PMID- 16262522 TI - A qualitative investigation of adolescents' experiences with parental HIV/AIDS. AB - The present qualitative study investigated the implications of parental HIV/AIDS for affected adolescents. The purpose of the study was (a) to understand adolescents' perceptions of and experiences with parental HIV/AIDS and (b) to explore how parental HIV/AIDS affects adolescents' psychosocial functioning, particularly in the home and school environments. Semistructured interviews were conducted with 9 adolescents between the ages of 11 and 17. Major themes included loss, transitions, disclosure, school implications, paradoxical situations, support networks, and coping. Findings validated and expanded on previous research and yielded hypotheses for future research. PMID- 16262523 TI - Completed suicide and psychiatric diagnoses in young people: a critical examination of the evidence. AB - Suicide rates of young people are increasing in many geographic areas. There is a need to recognize more precisely the role of specific mental disorders and their comparative importance for understanding suicide and its prevention. The authors reviewed the published English-language research, where psychiatric diagnoses that met diagnostic criteria were reported, to reexamine the presence and distribution of mental disorders in cases of completed suicide among young people worldwide. The number and geographical distribution of cases were limited (N = 894 cases). The majority of cases (88.6%) had a diagnosis of at least 1 mental disorder. Mood disorders were most frequent (42.1%), followed by substance related disorders (40.8%) and disruptive behavior disorders (20.8%). Those strategies focusing exclusively on the prevention and treatment of depression in young people need to be reconsidered. A comprehensive suicide prevention strategy among young people should target mental disorders as a whole, not depression alone, and consider contextual factors. PMID- 16262524 TI - The role of critical consciousness in multicultural practice: examining how its strength becomes its limitation. AB - Given the strengths of developing critical consciousness in multicultural practice, why do roadblocks persist? This article examines how the strengths of developing critical consciousness in multicultural practice can paradoxically become its limitation. Literatures from counseling psychology, clinical psychology, social psychology, social work, feminist theory, and critical theory are integrated in a discussion of the various components and strengths involved in working toward critical consciousness. This literature is then used to discuss some of the cognitive and affective limitations to achieving critical consciousness. Implications for practice are discussed. PMID- 16262525 TI - Antisense oligonucleotides and the rational design of new antitumor drugs. PMID- 16262526 TI - Antisense gene therapy using anti-k-ras and antitelomerase oligonucleotides in colorectal cancer. AB - AIM: To test the efficacy of anti-k-ras and antitelomerase oligonucleotides for disabling colorectal cancer cell growth. MATERIAL AND METHODS: An established human colorectal cancer cell line (SW 480, ATTC) was used. Oligodeoxiribonucleotides (ODNs) have a phosphorotioate modification to ensure intracellular intake. We used an antitelomerase ODN (Telp5) and two anti-k-ras ODNs (AS-KRAS and ISIS). AS-KRAS is designed to join the k-ras oncogene s exon 1. ISIS links to the terminal transcription unit 5 of k-ras. Telp5 joins the template region of the hTR telomerase subunit. ODNs have been tested in different concentrations (1, 5, 10, 20 micromolar). Cell viability has been tested at 48 and 72 hours. Statistical analysis and graphic design were made with the statistical package "Analyzing Data with GraphPad Prism-1999", GraphPad Sofware Inc., San Diego CA. We used the Student's t test for statistical analysis. RESULTS: The lowest dose (1 microM) was not effective. Using the highest dose (20 microM for 48 hours) of combined AS-KRAS and Telp5 cell viability decreased to 99.67%. The rest of results varied depending on ODN type, dose, and exposure time. CONCLUSIONS: Tested antisense ODNs stop colorectal cancer cell growth, and a combination of anti-telomerase and anti-k-ras is the most useful treatment. Efficacy is best with a higher dose and longer treatment period. PMID- 16262528 TI - Sequential treatment for proctalgia fugax. Mid-term follow-up. AB - INTRODUCTION: Proctalgia fugax (PF) is a benign, self-limiting disease characterized by episodes of intense anorectal pain at frequent intervals in the absence of organic proctological disease. Even though PF was described more than a century ago, its etiology remains unclear. Currently there is no information available. Few papers quoting many ways of management have been published. The aim of this study was to investigate patients complaining of this condition and to treat them with sequential therapy. PATIENTS AND METHODS: We devised a descriptive, prospective study of patients complaining of acute perianal pain- duration less than 30 minutes--without organic disease or previous perianal surgery since 1996 to 2002 in our Department. We treated these patients using a three-step treatment (1: information, hip bath, benzodiazepines; 2: sublingual nifedipine 10 mg, or topic 0.1% nitroglycerin on demand; 3: internal anal sphincterotomy if hypertrophy of the internal anal sphincter was demonstrated by anal ultrasonography and no improvement was confirmed with the previous steps of treatment). We defined remarkable improvement as a decrease in the number of episodes by half or in pain intensity by 50%. RESULTS: Fifteen patients with an average follow-up of 4 years. Anal endosonography confirmed a grossly thickened internal anal sphincter (IAS) in 5 cases. After the first step of treatment 7 patients improved and 1 patient was cured; after the second step of treatment 3 patients improved and 1 was cured; the third step was applied to 3 patients with a thickened IAS; 1 patient improved and 1 patient was cured. CONCLUSION: A total resolution of PF is not always possible, but we may improve symptoms and their frequency. Almost 50% of patients in our series improved with the first step of treatment; 30% of our patients had IAS hypertrophy. Anal endosonography can help in the diagnosis of organic diseases or IAS hypertrophy, for which we can perform an internal anal sphincter myectomy. PMID- 16262527 TI - Maintenance of T1 response as induced during PEG-IFNalpha plus ribavirin therapy controls viral replication in genotype-1 patients with chronic hepatitis C. AB - OBJECTIVES: To analyze the T1/T2 cytokine profile in CD8 T cells from peripheral blood mononuclear cells from patients with genotype-1 CHC during treatment with pegylated interferon (Peg-IFN) alpha2a plus ribavirin (RBV). To correlate Th1/Th2 balance with virological response. PATIENTS AND METHODS: In this prospective longitudinal study, a total of 28 naive genotype-1 CHC patients received Peg IFNalpha2a (180 microg/week) plus RBV (1-1.2 g/day) for 48 weeks. All patients (mean age 45 +/- 8 years) completed treatment and follow-up: 12 (43%) achieved a sustained virological response (SVR), 13 relapsed after end of treatment (47%), and only 3 (10%) were non-responders. Sixteen healthy controls were also analyzed (mean age 39 +/- 17 years). The production of IL-4, IFNgamma, and TNFalpha by CD8 T cells was measured by intracytoplasmic detection using flow cytometry in both resting and stimulated cells with a phorbol ester. STATISTICS: Student's t test for independent values, chi2 test, and ANOVA test were used; relapsers and non responders were joined to achieve a higher statistical power. RESULTS: At third month during treatment, phorbol ester-stimulated-IL-4 levels tend to be lower in patients who presented with SVR versus those who did not (0.97 vs 2.58; p = 0.1). No statistically significant differences were found in IFNgamma and TNFalpha levels at month 3. At EOT, the stimulated-IFNgamma production was significantly higher in patients with SVR (20 vs. 8; p < 0.05). Conversely, IL-4 production was higher in NR patients although these data did not reach statistical significance (p < 0.1). No significant differences were found in TNFalpha (14 vs. 7; p < 0.2). CONCLUSIONS: Cytokine T1 induced-response maintenance during combination treatment, measured as IFNgamma production by CD8+ T lymphocytes, is associated with SVR and suggests the replication control and later clearance of patients infected by genotype-1 HCV. PMID- 16262529 TI - Hyperhomocysteinemia and methylenetetrahydrofolate reductase 677C-->T and 1298A- >C mutations in patients with inflammatory bowel disease. AB - BACKGROUND: Hyperhomocysteinemia has been recently described in patients with inflammatory bowel disease (IBD), that could be related to the increased risk for thrombosis that exists in this disease. The aim of this study was the assessment of hyperhomocysteinemia in patients with IBD and its relation among vitamin B12 and folate levels, and methylenetetrahydrofolate reductase (MTHFR) 677C-->T and 1298A-->C mutations. PATIENTS AND METHODS: Fifty two consecutive patients with IBD were studied (29 women and 23 men); age: mean (standard deviation 41.7 [11.9] years) and 186 controls with no difference in age and gender. Hyperhomocysteinemia was considered as homocysteine levels higher than mean plus two standard deviations of the control group (> or = 13 micromol/l). RESULTS: patients had an elevated prevalence of hyperhomocysteinemia (17.3 vs. 3.7%; p = 0.002) and lower folate (7.6 [4.1] vs. 8.9 [3.7] ng/ml; p = 0.01) and B12 vitamin levels (499 [287] vs. 603 [231] pg/ml; p = 0.003). Homocysteinemia was higher (14.3 [5.8] vs. 9.1 [3.9] micromol/l; p = 0.006) in 6 patients (11.5%) that had suffered thromboembolism. Frequency of MTHFR 677C-->T (13.5 vs. 11.3%; p = 0.66) and 1298A-->C (7.8 vs. 7.0%; p = 0.76) mutations was not increased in patients. Odds ratio (OR) for IBD in hyperhomocysteinemic patient was 5.51, 95% confidence interval (CI), 1.81-16.76; p = 0.002). Hyperhomocysteinemia was negatively associated with feminine gender (OR 0.08, 95% CI 0.01-0.49; p = 0.006) and folate levels (OR 0.04, 95%CI: 0.007-0.20; p < 0.001). CONCLUSIONS: hyperhomocysteinemia is associated with IBD and low folate levels, and could be involved in development of thromboembolism. MTHFR 677C-->T and 1298A-->C mutations are not related with the disease. PMID- 16262530 TI - Helicobacter pylori infection and gastric mucosal epithelial cell apoptosis. PMID- 16262531 TI - Association between Whipple's disease and Giardia lamblia infection. AB - Whipple' disease is mainly characterized by affecting the digestive system, although it can be a multisystemic process with different clinical symptoms. The bacillus causing the disease has been isolated and cultivated in 2000 and the genome sequence has been recently analyzed in 2003, which means new perspectives for its diagnosis and treatment. Giardiasis is an infestation caused by a protozoo and may cause a malabsorption syndrome or run in a subclinic way. The case of a middle-aged male is described, who after a three-year period of migratory arthralgias, showed weight loss, diarrheas and abdominal pain, being diagnosed of Giardiasis, and after the persistent symptoms and a number of studies, was diagnosed with Whipple disease. Nineteen cases of Giardia-Whipple coinfection have been described in the literature, but the reason of this association has not been found yet. The discussion on whether there is an alteration in the immunitary system which facilitates infections or, the development of an infection lead to the other one, goes on. PMID- 16262532 TI - Hypertrophic myopathy of the internal anal sphincter: a rarely recognized cause of proctalgia. PMID- 16262533 TI - [Treatment for hepatitis C virus associated cryoglobulinemia with interferon alpha and ribavirin]. PMID- 16262534 TI - [Anisakis simplex on an ulcer in a Billroth II patient]. PMID- 16262535 TI - [Esophagus actinomycosis causing upper gastrointestinal bleeding]. PMID- 16262537 TI - [Massive pneumatosis intestinalis associated with enteral nutrition and needle catheter]. PMID- 16262536 TI - [Right parametric mullerian cyst]. PMID- 16262538 TI - [Intestinal perforation: an unusual complication of the intestinal tuberculosis that simulated Crohn's disease]. PMID- 16262539 TI - Predictors of cigarette use among South African adolescents. AB - This study assessed the interrelation among domains of ethnic factors; the individual's sense of well-being; personality, attitudes, and behaviors; sibling and peer smoking; and adolescent smoking behavior. The sample consisted of 1,468 South African adolescents selected from 4 ethnic groups self-identified as defined by current South African usage: Black (mainly Zulu and Xhosa), Indian, White, and Colored (mixed ancestry). In accordance with family interactional theory, there was a sequence of patterning from ethnic factors and the individual's sense of well-being to adolescent personality, attitudes, and behaviors and models of smoking. All of the 4 domains in the model also had a direct effect on adolescent smoking behavior. The findings suggest 4 possible targets of therapeutic or preventive intervention with regard to adolescent smoking: ethnic factors; the individual's sense of well-being; personality, attitudes, and behaviors; and smoking within the peer group. PMID- 16262540 TI - Cognitive behavioral stress management effects on mood, social support, and a marker of antiviral immunity are maintained up to 1 year in HIV-infected gay men. AB - Numerous herpesvirus infections are associated with clinically relevant outcomes as well as an accelerated HIV replication rate and subsequent disease progression. Stress management interventions may improve markers of cellular immune control over latent herpesvirus infections and these changes appear to be mediated by perceptions of increased social support availability. We examined the effects of a group-based cognitive behavioral stress management (CBSM) intervention on distress, dysphoria, perceived social support, and herpesvirus immunoglobulin G (IgG) antibody titers during the 6 to 12 months following the intervention. Of those who were initially randomized, 49 HIV-infected men were followed during the 6- to 12-month period after randomization to either a 10-week CBSM intervention (n = 31) or a modified wait-list control condition (n = 18). Measures of distress, dysphoria, social support, and blood samples for herpesvirus IgG titers were taken at baseline, immediately following CBSM and at 6- to 12-month follow-up. Men in CBSM displayed maintenance of previously observed intervention effects on dysphoria, reliable alliance support, and herpesvirus IgG antibody titers (i.e., Epstein-Barr virus capsid antigen; EBV VCA). Intervention-related changes in EBV-VCA were unrelated to changes in lymphocyte subsets (i.e., CD4+, CD8+, and CD4+:CD8+) or changes in measures of dysphoria and social support during the investigation period. Data indicate that HIV-infected men participating in a CBSM intervention maintain better psychosocial status and immunologic control of latent EBV infection up to 1 year after its conclusion. PMID- 16262541 TI - Emerging adulthood and patterns of physical activity among young Australian women. AB - The transition from adolescence to young adulthood is associated with a sharp decline in physical activity, particularly for women. This article explores the relations between physical activity status and change and status and change in four life domains: residential independence, employment status, relationship status, and motherhood. Two waves of survey data from a representative sample of 8,545 Australian women, aged 18-23 at Survey 1 and 22-27 at Survey 2, were analyzed. Cross-sectionally, physical inactivity was most strongly related to being a mother, married, and not being in the labor force. Longitudinally, decreases in physical activity were most strongly associated with moving into a live-in relationship, with getting married, and with becoming a mother. When considered in combination, women who were married with children and not employed outside the home were the most likely to be physically inactive. The data suggest that adoption of adult statuses, particularly traditional roles involving family relationships and motherhood, is associated with reductions in physical activity for these women, although it is possible that the effect is driven by socioeconomic factors associated with early transitions. The data suggest a need for interventions to promote continued physical activity among young women who cohabit or marry and among those not in the workforce, in addition to those supporting young mothers to be physically active. PMID- 16262542 TI - Depressive symptoms and life satisfaction in elderly women are associated with natural killer cell number and cytotoxicity. AB - Well-preserved natural killer (NK) cell cytotoxicity (NKCC) is associated with healthy aging. The objective of the survey was to investigate psychological factors related to NKCC and NK cell populations in elderly women. A cross sectional study involving 181 participants was conducted using the Japanese version of the 28-item General Health Questionnaire (GHQ) and additional questions assessing psychological status and lifestyle. Spearman's rank test revealed a significant negative correlation between NKCC and the GHQ depression subscale (GHQ-D) scores. Significantly reduced NKCC was found in participants presenting high GHQ-D scores (12 < or = GHQ-D, n = 58) compared with those showing middle (8 < or = GHQ-D < or = 11, n = 55) or low (GHQ-D = 7, n = 68) scores. Adjusting for covariates regarding lifestyle, multiple logistic regression analysis was applied; consequently, significant associations were found between reduced NKCC and high depressive symptoms and between increased NK cell numbers and life satisfaction. These results indicated a clue to longitudinal studies in the future. PMID- 16262543 TI - Long-term effects of two psychological interventions on physical exercise and self-regulation following coronary rehabilitation. AB - In cardiac rehabilitation programs, patients learn how to adopt a healthier lifestyle, including regular, strenuous physical activity. Long-term success is only modest despite good intentions. To improve exercise adherence, a 3-group experiment was designed that included innovative psychological interventions. All 3 groups underwent a standard care rehabilitation program. Patients in the 2 treatment groups were instructed not only to produce detailed action plans but also to develop barrier-focused mental strategies. On top of this, in 1 of these groups a weekly diary was kept for 6 weeks to increase a sense of action control. At the end of a standard cardiac rehabilitation program, 240 patients were randomly assigned to these treatment groups plus a standard care control group. Treatments resulted in more physical activity at follow-up and better adherence to recommended levels of exercise intensity. Moreover, self-regulatory skills such as planning and action control were improved by the treatments. Follow-up analyses demonstrated the mediating mechanisms of self-regulatory skills in the process of physical exercise maintenance. Findings imply that interventions targeting self-regulatory skills can enable post-rehabilitation patients to reduce behavioral risk factors and facilitate intended lifestyle changes. PMID- 16262544 TI - A 5-year follow-up study of aggression at work and psychological health. AB - In a longitudinal cohort study, organizational climate and long-term effects of exposure to nasty teasing (aggression) at work were investigated. The baseline consisted of a representative sample of Danish employees in 1995 with a response rate of 80% (N = 5,652). Of these, 4,647 participated in the follow-up in 2000 (response rate 84%). In 1995, 6.3% were subjected to nasty teasing with no significant gender difference. At baseline, we found significant associations among nasty teasing, a negative organizational climate, and psychological health effects. In the follow-up analyses, associations were found between exposure to nasty teasing at baseline and psychological health problems at follow-up, even when controlled for organizational climate and psychological health at baseline and nasty teasing at follow-up. Stratified for gender, the follow-up associations were significant for women but not for men. Low coworker support and conflicts at baseline and teasing at follow-up mediated the effects on men. PMID- 16262545 TI - Self-efficacy and motivation to quit during participation in a smoking cessation program. AB - The associations between failure to quit and posttreatment self-efficacy and motivation were examined among 600 African American smokers enrolled in a randomized trial testing the efficacy of bupropion for smoking cessation. Participants also received brief motivational counseling and were followed for 6 months. Baseline levels of self-efficacy and motivation for all participants were high (8.2 and 8.5 on a 10-point scale, respectively). Longitudinal analyses indicated that smokers who failed to quit were less likely than quitters to report high self-efficacy and motivation from posttreatment to follow-up. However, examination of mean self-efficacy and motivation scores at posttreatment and follow-up revealed that smokers continued to sustain high self-efficacy and motivation. Mean self-efficacy and motivation scores differed by less than 1 point from baseline levels, even though the majority of participants failed to quit smoking. Results suggest that unsuccessful participation in a smoking cessation program does not meaningfully reduce smokers' self-efficacy and motivation to quit. PMID- 16262546 TI - Self-efficacy: a predictor for smoking cessation contemplators in Kuwaiti adults. AB - The failure of most of the smoking cessation programs might be due to negligence of including self-efficacy as an imperative factor in changing many adverse behaviors such as smoking. This study investigates the role of self-efficacy as a predictor for smoking cessation contemplators and precontemplators in adult male Kuwaiti smoker employees. A sample of 657 Kuwaiti male smokers represented the target population. Factor analysis with varimax rotation to the self-efficacy 16 items scale revealed four essential factors-mood changes, relaxation, stress, and self-image-for smoking urge in the studied population. Contemplators had significant higher mean self-efficacy total scores and Factor IV (self-image) subtotal scores than precontemplators. Stepwise multivariate logistic binary regression analysis illustrated that self-efficacy is the first predictor for contemplating smoking cessation followed by monthly income. Self-efficacy as a cognitive determinant should be considered to mediate improvement in the smoking cessation programs. PMID- 16262547 TI - Impact of mindfulness-based stress reduction (MBSR) on sleep, mood, stress and fatigue symptoms in cancer outpatients. AB - Sleep disturbance is a very common problem for cancer patients that has largely not been addressed in the clinical intervention literature. Mindfulness meditation has demonstrated clinical benefits for a variety of patient populations in other areas of functioning. This study examined the effects of an 8-week Mindfulness-Based Stress Reduction (MBSR) program on the sleep quality of a heterogeneous sample of 63 cancer patients. Overall sleep disturbance was significantly reduced (p < .001) and participants reported that their sleep quality had improved (p .001). There was also a significant reduction in stress (p < .001), mood disturbance (p = .001), and fatigue (p < .001). The associations among these changes and implications for improving quality of life of cancer patients are discussed. PMID- 16262549 TI - Evaluation of student performance in combined baccalaureate-MD degree programs. AB - BACKGROUND: Among the innovations in medical education during the 1960s and early 1970s was the emergence of combined baccalaureate-MD degree programs. Viewed as educational experiments, an evaluation of performance outcomes for these programs is needed. SUMMARY: Performance outcomes of students in combined-degree programs, reported in the literature 1966-1996, are reviewed. Attrition rates are lower for combined-degree students than for traditional premedical students, and there are usually no differences in attrition between combined-degree and traditional students in the medical phase of their education. All measures of performance indicate that students in combined-degree programs achieve a level of competency comparable to traditional medical students. CONCLUSIONS: The support systems of the Baccalaureate-MD degree programs facilitate retention in a medical career. The educational experiment of the combined-degree programs has demonstrated that future physicians can be successfully selected from high school. PMID- 16262551 TI - How do faculty evaluate students' case presentations? AB - BACKGROUND: Although the case presentation is a frequent activity, little is known about the attributes that faculty members use when assessing case presentation ability. PURPOSE: Define the dimensions used by faculty when assessing students' case presentation abilities. METHODS: Eleven internal medicine faculty members independently assessed the same 17 videotaped student case presentations. Raters used an evaluation form assembled with 4 descriptors of content and 6 attributes of communication style. For each evaluator, correlation analyses were used to examine associations among the individual descriptors and a student's overall evaluation. RESULTS: Despite efforts toward uniform scoring, faculty evaluators varied in their application of rating criteria. The majority of raters had significant (p < .01) correlation between 4 of the 10 dimensions and the overall score. These 4 were history of present illness and three communication skills: economy, fluency, and precision of language. CONCLUSIONS: Faculty members' assessment of case presentations is based on both the facts presented and communication style. These identified performance dimensions can facilitate teaching students to present cases and allow more informative feedback on their abilities. PMID- 16262550 TI - A longitudinal investigation of medical student attitudes toward the medically indigent. AB - BACKGROUND: Studies have shown that medical student attitudes decline as students progress through school. Controversy persists about the meaning and generalizability of these findings because studies have not been longitudinal and many were conducted prior to an influx of female students. Much of medical education occurs in academic medical center environments where disproportionate numbers of indigent (uninsured and underinsured) patients seek care. This study examined whether students' attitudes toward providing care to indigent clientele changed over time. PURPOSE: To track longitudinal changes in attitudes toward providing care for the medically indigent of 1 cohort of medical students and to determine if gender differences existed and persisted over time. METHODS: Students entering a Southwest medical school in 1989 participated in this study. Attitudes were compared using the Medical Students' Attitudes Toward the Underserved questionnaire, a self-report, attitudinal scale. RESULTS: Student attitudes declined during the 1st year of school, remained fairly stable during Years 2 and 3, and declined further during Year 4. Analyses comparing 1st- and 4th-year students showed a decline in attitudes for men and women for each scale except Basic Services. Although both women's and men's attitudes declined, women's attitudes remained more favorable across the 4 years. CONCLUSIONS: A clearer understanding of how female medical students sustain socially responsible attitudes throughout medical school may help educators develop support mechanisms for men and women that would promote an ethic of social responsibility and encourage students to work with indigent clientele. PMID- 16262552 TI - The effect of a standardized patient instructor experience on students' anxiety and confidence levels performing the male genitorectal examination. AB - BACKGROUND: Standardized patient instruction (SPI) is recommended as a nonthreatening method for teaching the male genitorectal examination. The article's purpose is to describe the method's effectiveness in reducing anxiety and increasing confidence among men and women students from diverse cultures. DESCRIPTION: We implemented an SPI program in 1993 to teach the examination to 2nd-year students. Students performed their examinations in mixed gender groups of three; a man student was responsible for disrobing the SPI and performing the first exam. During the 45-min session, each of the three students performed the examination in turn, and each received immediate feedback on their technique and interpersonal approach to the patient. EVALUATION: All students (n = 190) evaluated their SPI encounter immediately after the session had ended. CONCLUSIONS: Men and women students from all ethnic groups reported decreased anxiety and increased confidence levels after the SPI session. These findings indicate that a carefully orchestrated SPI session is effective in reducing students' anxiety about crossing personal space boundaries, overcoming a variety of proscriptions on gender-appropriate interactions, and increasing their confidence to perform this sensitive examination. PMID- 16262553 TI - Exploratory study of residents' conceptual framework for critical appraisal of the literature. AB - BACKGROUND: Limited success of efforts to improve residents' critical appraisal skills may be due to poor understanding of their preintervention conceptual framework. DESCRIPTION: To describe internal medicine residents' a priori conceptual approach to critical appraisal of intervention trials, we performed an exploratory content analysis of 41 residents' unprompted descriptions of strengths and weaknesses of a randomized trial of spa therapy for back pain. EVALUATION: Eighty-eight percent of responses were assigned independently by 2 reviewers to 23 categories ( kappa .65). Residents agreed on some important characteristics (e.g., blinding), disagreed on others (e.g., similarity of treatment and control groups), frequently mentioned some irrelevant characteristics (e.g., "objective" vs. "subjective" outcomes), and rarely mentioned some important criteria (e.g., intention to treat). CONCLUSIONS: An open-ended questionnaire reliably revealed both expected and unexpected conceptions and misconceptions among residents who had received minimal or no formal instruction in critical appraisal. We propose a classification model for residents' critical appraisal concepts that can be tested in future studies and used to identify areas for focusing interventions to improve resident physicians' critical appraisal skills. PMID- 16262554 TI - Commitment to the underserved: evaluating the effect of an extracurricular medical student program on career choice. AB - BACKGROUND: This study was conducted to compare the practice locations and characteristics of physicians who participated as medical students in an extracurricular program to foster interest in careers of service to medically underserved populations with those of their classmates who did not participate in the program. METHODS: Using a mailed questionnaire, we conducted a cross sectional study of graduates from the classes of 1983-1987 at one southwestern, public medical school. All Commitment to Underserved People (CUP) participants (n = 94) and a random sample of nonparticipating classmates (n = 188) were surveyed. CUP is an extracurricular project with components in each of 4 years of medical school that provides peer and faculty support, curriculum enrichment, and direct service to medically underserved populations. Outcome measures included the size of community of practice, practice type, and practice patient characteristics. RESULTS: Sixty seven (71%) of CUP participants and 126 (67%) of nonparticipants responded. CUP participants were more likely to be women, to specialize in family practice, to practice in the Indian Health Service (IHS) or overseas, to be located in a community of 25,000 or less, and to have participated in the state, service-payback loan program. In multiple regression, the specialty of family practice was associated with practice in a small community, the IHS, and a community health center; CUP participation was correlated with practice in small communities, the IHS, and a foreign country. CONCLUSIONS: Participation in the CUP program was associated with the specialty choice of family practice and with practice in settings associated with medically underserved populations. The CUP program has been successful in sustaining entering medical students' interests in underserved practice. PMID- 16262558 TI - Emerging anti-inflammatory agents for allergic rhinitis. AB - Allergic rhinitis (AR) is a high-prevalence disease, affecting 10-15% of the general population. AR is sustained by an IgE-mediated reaction, and by a complex inflammatory network of cells, mediators and cytokines that becomes chronic when exposure to allergen persists. A T helper 2 (TH2)-biased immune response is the basis for the allergic inflammation. The current therapeutic strategy is mainly based on drugs (antihistamines, nasal corticosteroids, cromones, decongestants) and allergen immunotherapy. Drugs are overall effective in controlling symptoms, but do not modify the immune background that leads to allergic inflammation, and safety concerns may be present especially for prolonged treatments. Immunotherapy can modify the allergic response, but there is still space for improvement. Nowadays, several approaches are under investigation to optimise the management of AR. On one hand, new drugs and antimediators are being developed; on the other hand, attempts are made to selectively block relevant signal pathways of allergic reaction. Finally, one of the major goals is to modify the TH2-biased immune response by improving the characteristics and modes of action of allergen immunotherapy. PMID- 16262559 TI - Emerging drugs for idiopathic pulmonary fibrosis. AB - Pulmonary fibrosis is often the end stage of chronic, persistent, low-level lung injury, either of known or unknown cause. The most severe form of pulmonary fibrosis is idiopathic pulmonary fibrosis (IPF), a disease process of unknown aetiology and one that often leads to respiratory failure and death. At present there are no proven or effective drug therapies for IPF. Recent advances in understanding of disease pathogenesis have focused attention on drug targeting of fibrogenic pathways, as opposed to traditional anti-inflammatory approaches. In this report, the present status of drug development of a number of emerging antifibrotic strategies and agents that may prove more effective in the therapy of this progressive, debilitating and fatal disease are reviewed. PMID- 16262560 TI - Targeting the renin-angiotensin system for the reduction of cardiovascular outcomes in hypertension: angiotensin-converting enzyme inhibitors and angiotensin receptor blockers. AB - Agents that counteract the negative impact of the renin-angiotensin-aldosterone system (RAAS) are effective antihypertensives and reduce the risk of developing Type 2 diabetes. Contrary to common perception, angiotensin-converting enzyme inhibitors do not share the apparent benefit of angiotensin II receptor blockers (ARBs) in reducing risk of cardiovascular-disease outcomes, particularly stroke, in randomised clinical trials. RAAS agents, especially ARBs, are well tolerated. Use of ARBs alone or in combination with other classes of antihypertensive agents to lower blood pressure and/or medications to control other conditions (e.g., insulin sensitivity) reduces risk of cardiovascular disease outcomes and Type 2 diabetes with excellent tolerability. Selected issues related to use of RAAS agents as antihypertensive therapies (e.g., Type 2 diabetes, global risk management, multiple drug therapy and coronary heart disease) are addressed. PMID- 16262557 TI - The unexpected side effects of new nonsteroidal anti-inflammatory drugs. AB - Conventional nonselective NSAIDs are classically associated with a risk of gastrointestinal disorders. These drugs have a broad range of relative selectivity towards the COX family, mainly towards two isoforms of these enzymes: COX-1 and -2. As examples, ketorolac, flurbiprofen, ketoprofen and indomethacin have increased COX-1 selectivity when compared with naproxen and ibuprofen. PMID- 16262561 TI - Emerging drugs for diabetic nephropathy. AB - There is an increasing number of patients with diabetes mellitus in many countries. Diabetic kidney disease, one of its microvascular complications, is also increasing markedly and has become a major cause of end stage renal disease worldwide. Intervention for preventing and delaying the development and progression of diabetic kidney disease is not only a medical concern, but also a social issue. Despite extensive efforts, however, medical interventions thus far are not effective enough to prevent the progression of the disease and the development of end stage renal disease. This justifies attempts to develop novel therapeutic approaches for diabetic nephropathy. Recent insights on its pathogenesis and progression have suggested new targets for the specific treatment of this disease. They include aldosterone, aldose reductase, arachidonic acid metabolites, growth factors, advanced glycosylation end products, peroxisome proliferator-activated receptors and endothelin. Several other biochemical mediators have been targeted in experimental animal models with the goal to prevent diabetic nephropathy progression, but translation to clinics of these experimental achievements are still limited or lacking. PMID- 16262563 TI - Emerging therapies in multiple sclerosis. AB - Multiple sclerosis (MS) is the most common neurological cause of disability in young people. The disease-modifying treatments, IFN-beta and glatiramer acetate, have been widely available over the last decade and have shown a beneficial effect on relapse rate and magnetic resonance imaging parameters of disease activity; however, their effect on disease progression and disability is modest. Therefore, the search for alternative treatment strategies continues. As understanding of the heterogeneous pathophysiology of MS has increased, emphasis has shifted to more selective therapy that targets components of the inflammatory cascade and the promotion of remyelination and neuroprotection. These agents target the blood-brain barrier, systemic immune dysfunction, local inflammation and neurodegeneration. Combination therapies are being investigated for patients who fail first-line treatments. Many new drugs are being developed and tested that address these issues with the aim of finding a more effective and convenient therapy. These include humanized monoclonal antibodies such as daclizumab (IL-2 antagonist), oral immunomodulators such as sirolimus and statins and neuroprotective agents such as NMDA antagonists and Na+-channel blockers. Many of the treatments discussed in this review are still at early stages of development, but provide exciting potential treatment options; others have proved disappointing in larger extended-phase studies. PMID- 16262562 TI - Emerging drugs for renal cell carcinoma. AB - Renal cell carcinoma (RCC) still represents a therapeutic challenge when patients have advanced or metastatic disease. Treatment using IL-2 and IFN-alpha continues to be the standard of care in patients who are able to tolerate such regimens. Targeted therapy may become the first-line treatment for patients resistant or intolerant to cytokines as new emerging drugs continue to be investigated. Understanding the genetic abnormalities related to the development of RCC (e.g., VHL gene abnormalities) and identifying molecular targets (e.g., epidermal growth factor, vascular endothelial growth factor and carbonic anhydrase IX) are playing a major role in the emergence of these novel agents for the treatment of this malignancy. Overall, these drugs are better tolerated and more acceptable to use by patients than the traditional cytokine-based regimens. The use of oral drugs to treat various malignancies including RCC seems to be the new paradigm of the future. Further understanding of their mechanisms of action and confirmation of their benefits on the clinical outcome is needed. PMID- 16262564 TI - Acetylcholinesterase inhibitors for the treatment of dementia in Alzheimer's disease: do we need new inhibitors? AB - Acetylcholinesterase inhibitors (AChEIs) have been shown to produce a small, but significant, improvement in cognition in patients with Alzheimer's disease. However, not all patients respond equally, and cognitive benefit may be of limited duration. Although new AChEIs continue to be developed, more recent studies have been aimed at developing inhibitors that have additional actions separate from AChE inhibition. Importantly, new treatments that target the underlying pathogenic mechanism of Alzheimer's disease (statins, secretase inhibitors, vaccination) may eventually emerge. These new treatments could make AChEI therapy less relevant for treatment of Alzheimer's disease. PMID- 16262565 TI - Glutamatergic neurotransmission modulators as emerging new drugs for schizophrenia. AB - Schizophrenia is a neurodevelopmental mental disorder whose aetiology includes genetic and environmental factors. Because of its early onset, chronicity and characteristic interference with education, employment and socialisation, this illness represents a tremendous human and economic burden to those who suffer from it, their families and society as a whole. Conventional and atypical antipsychotics, which mainly affect dopaminergic and serotonergic neurotransmission, are currently the cornerstone of schizophrenia treatment. Although the introduction of atypical antipsychotics represents a major development and, overall, antipsychotics are efficacious against psychotic symptoms, there remains a critical unmet need for innovative medications with improved efficacy and tolerability for the negative symptoms and cognitive deficits associated with schizophrenia. These dysfunction domains are reliable predictors of long-term disability and treatment outcome and are presently viewed as crucial targets for new pharmacological treatments of schizophrenia. Within this medication development framework, the modulation of glutamatergic neurotransmission has become the focus of intense research. Glutamate (GLU) mediated neuronal processes are critical throughout the brain and glutamatergic neurotransmission dysfunctions have been hypothesised to play a crucial role in schizophrenia pathophysiology. Glutamatergic neurotransmission may be modulated at multiple levels, with GLU receptor families and their subtypes representing a modulatory site-rich environment for drug research. Numerous types of neurotransmission modulators, acting at the NMDA, AMPA and metabotropic GLU receptors, and/or affecting GLU synaptic release, are hypothesised to be beneficial for schizophrenia treatment, and are presently in various stages of development. For some of these compounds, preliminary studies have furnished encouraging clinical data. Ongoing and planned research is expected to provide, in the near future, critical information regarding the practical utility and tolerability of glutamatergic approaches for schizophrenia pharmacotherapy. PMID- 16262566 TI - Emerging targeted treatments for malignant glioma. AB - This paper focuses on the medical management of malignant gliomas, which is currently undergoing change. It suggests that as surgery and radiotherapy are of limited benefit in the treatment of these tumours, medical therapies may have the potential to offer a better alternative. The current therapies for glioma and the targeted agents in clinical trials are reviewed. There is a general acceptance that temozolomide in combination with radiotherapy is replacing radiotherapy alone as first-line therapy in high-grade astrocytic gliomas. Within the realms of clinical research, it can be seen that there is a shift away from therapies targeting the end effect of deregulated cell-cycle control, to targeting specific and individual genetic aberrations in tumours. Finally, the paper questions current clinical trial methodology and tentatively suggests ways in which this may be improved. PMID- 16262567 TI - Epidermal growth factor receptor inhibitors in the treatment of non-small cell lung cancer. AB - Non-small cell lung cancer (NSCLC) is the leading cause of cancer-related death in both men and women. Despite the introduction of the newer cytotoxic agents in NSCLC treatment during the last decade the survival rates of patients have reached a plateau. New strategies are clearly needed to improve treatment outcomes. Epidermal growth factor receptor (EGFR) has a key role in cancer development and progression and has been recognised as a target of increasing importance in NSCLC. Gefitinib, erlotinib and cetuximab are EGFR-targeting agents that are being extensively evaluated in NSCLC. EGFR inhibitors demonstrate significant clinical activity in approximately 10-20% of pretreated NSCLC patients. Somatic mutations in the kinase domain of the receptor have been shown to be associated with enhanced sensitivity to EGFR inhibitors. However, four large Phase III randomised, placebo-controlled trials of gefitinib and erlotinib in combination with standard platinum-based first-line chemotherapy failed to show any survival benefit in patients receiving the study drugs. Possible reasons include patient selection, drug scheduling, trial design or other factors. Active research is ongoing to improve the efficacy of EGFR inhibitors as monotherapy or in combination with other treatment modalities. PMID- 16262569 TI - Current and emerging therapies for the lysosomal storage disorders. AB - Targeted treatments for the lysosomal storage disorders (LSDs), in the form of enzyme replacement and/or substrate depletion, have been shown to be relatively safe and effective in reversing core disease features in selected clinical subtypes (including Gaucher disease types I and III, Fabry disease and the Hurler Scheie syndrome). These approaches have expanded the therapeutic options available to patients with rare genetic disorders, beyond palliative measures (such as liver or kidney transplantation for end-organ failure) and cellular replacement through bone marrow transplantation. Present efforts are focused on the development of novel strategies, including chaperone-mediated enzyme enhancement and genetically engineered stem cell therapy. In the coming decades, a broadening therapeutic horizon for patients with inborn errors of metabolism is anticipated, and the growing experience in the management of patients with LSDs will serve as an instructive model. Among the many challenges will be determination of the extent to which these therapies have modified the course of disease beyond merely extending the age of survival, but also enabling a meaningful patient quality of life, and the minimisation of current resource use. The projected lifetime acquisition costs of newly introduced therapeutic options also raises several issues, related to equitable access and the large opportunity costs for other therapeutic areas, that will need to be addressed by healthcare policy makers and third-party payers. PMID- 16262568 TI - Treatment strategies for acromegaly. AB - Acromegaly is a chronic debilitating disorder caused by a growth hormone (GH) producing pituitary adenoma. Active acromegaly is associated with a two- to fourfold increased mortality risk, mainly from cardiovascular disease. Transsphenoidal surgery is considered as the treatment of choice because of the rapidity of cure and normalisation of survival. Secondary treatment modalities are radiotherapy and medical treatment, and are important because surgery in the best hands cures only approximately 60% in long-term studies. Medical treatment with slow-release formulations of somatostatin are now widely used, also as primary treatment, and appear to be safe and effective in 50-60% of the patients. However, no data on mortality risk with these drugs is available. Recently, a GH receptor blocking agent, pegvisomant, was licensed for use in acromegaly and appears to normalise IGF-1 in almost all patients. This article examines the pathophysiology of acromegaly, currently used medicines and their safety and efficacy, and the new drugs that are in development. PMID- 16262572 TI - Pure or tarnished: are systematic reviews blind or biased? PMID- 16262573 TI - Can Cochrane Reviews in controversial areas be biased? A sensitivity analysis based on the protocol of a Systematic Cochrane Review on low-level laser therapy in osteoarthritis. AB - OBJECTIVE: The aim of this study was to test if a conclusion in a systematic review of low-level laser therapy (LLLT) for osteoarthritis from the Cochrane Library was valid and robust. BACKGROUND DATA: Health policy decisions often rely on conclusions from the Cochrane Database of Systematic Reviews for approval of new therapies, although their validity for controversial non-pharmacological treatment has been questioned. METHODS: Validity was tested against a nine-item checklist for systematic reviews. Review selections were analyzed for possible discrepancies between trial and review reports, and omissions of relevant trials and data. Alternative data from discrepancies and omissions were then imputed in a sensitivity analysis, to test if review conclusions were robust. RESULTS: Only clinicians who had performed LLLT trials with negative results were invited into the review group. Review quality was sound in areas of literature search and methodological assessments, and some of the limitations were mentioned. The statistical analysis held 18 questionable selections such as omissions of trials, data, and subgroup analyses. These selections systematically favored the negative review conclusion. Without altering the review protocol, the sensitivity analysis of combined results changed to significantly positive for continuous and categorical data when data from all included trials were combined. Further sensitivity analyses with inclusion of valid non-included trials, performance of missing follow-up, and subgroup analyses revealed consistent and highly significant results in favor of active LLLT. CONCLUSIONS: In this example, the Cochrane review conclusion was neither robust nor valid. Representation of experts and different views on efficacy in the review group and extensive use of sensitivity analyses could probably improve quality control of reviews in areas of controversy. PMID- 16262574 TI - Low-power laser treatment for shoulder pain. AB - OBJECTIVE: The objective of this study is to investigate the effect of low-power gallium-arsenide laser treatment on the patients with shoulder pain. BACKGROUND DATA: Low-energy laser therapy has recently been popularized in the treatment of various rheumatologic, neurologic, and musculoskeletal disorders such as osteoarthritis, rheumatoid arthritis, fibromyalgia, carpal tunnel syndrome, rotator cuff tendinitis, and chronic back pain syndromes. METHODS: A total of 40 patients who applied to our clinic with shoulder pain and complied with the selection criteria were included in the study. The patients were randomly assigned into Group I (n = 20, laser treatment) and Group II (n = 20, control). In Group I, patients were given laser treatment and an exercise protocol for 10 sessions during a period of 2 weeks. Laser was applied over tuberculum majus and minus, bicipital groove, and anterior and posterior faces of the capsule, regardless of the existence of sensitivity, for 1 min at each location at each session with a frequency of 2000 Hz using a GaAs diode laser instrument (Roland Serie Elettronica Pagani, wavelength 904 nm, frequency range of 5-7000 Hz, and maximum peak power of 27 W, 50 W, or 27 x 4 W). In Group II, placebo laser and the same exercise protocol was given for the same period. Patients were evaluated according to the parameters of pain, palpation sensitivity, algometric sensitivity, and shoulder joint range of motion before and after treatment. RESULTS: Analysis of measurement results within each group showed a significant posttreatment improvement for some active and passive movements in both groups, and also for algometric sensitivity in Group I (p < 0.05-0.01). Posttreatment palpation sensitivity values showed improvement in 17 patients (85%) for Group I and six patients (30%) for Group II. Comparison between two groups showed superior results (p < 0.01 and p < 0.001) in Group I for the parameters of passive extension and palpation sensitivity but no significant difference for other parameters. CONCLUSIONS: The results of our study have shown better results in palpation sensitivity and passive extension, but no significant improvement in pain, active range, and algometric sensitivity in laser treatment group compared to the control group in the patients with shoulder pain. PMID- 16262575 TI - Comparative study of root canals instrumented manually and mechanically, with and without Er:YAG laser. AB - OBJECTIVE: The aim of this study was to compare the scanning electron microscopic (SEM) morphology of root canal walls instrumented manually and mechanically with and without the application of Er:YAG laser. METHODS: Single-rooted, human, extracted teeth were cut at the cementoenamel junction. The teeth were divided into four groups: teeth that were simply instrumented manually, those that were instrumented manually and laser treated, a group that was mechanically instrumented, and a fourth in which the teeth were mechanically instrumented and laser treated. After instrumentation, the teeth were split longitudinally to facilitate their evaluation by SEM. RESULTS: The results show that the use of the Er:YAG laser is effective in removing the smear layer from root canal walls. CONCLUSIONS: The removal of the smear layer is essential to the success of endodontic treatment; the use of the Er:YAG laser combined with rotary and manual techniques improves the cleanliness of root canals. PMID- 16262576 TI - Effect of In-Ga-Al-P diode laser irradiation on angiogenesis in partial ruptures of Achilles tendon in rats. AB - OBJECTIVE: This study was conducted to analyze the effect of different irradiances of low-level laser therapy (LLLT) on angiogenesis after partial rupture of Achilles tendon of rats. BACKGROUND DATA: METHODS: Ninety-six animals were divided into three groups subject to treatment during 3, 5, and 7 days post lesion. Thirty-two animals were used in each group. The groups were further divided into four subgroups with eight animals in each, receiving In-Ga-Al-P laser (660 nm) treatment at (1) mean output of 10 mW, (2) 40 mW during 10 sec, (3) a sham subgroup, and (4) a non-treatment subgroup. Each animal was subjected to a lesion of the Achilles tendon by dropping a 186-g weight from a 20-cm height over the tendon. Treatment was initiated 6 h post-injury for all the groups. Blood vessels were colored with India ink injection and were examined in a video microscope. RESULTS: Laser exposure promoted an increase in blood vessel count when compared to controls. The 40-mW group showed early neovascularization, with the greatest number of microvessels after three laser applications. The 10-mW subgroup showed angiogenesis activity around the same time as the sham laser group did, but the net number of vessels was significantly higher in the former than in the controls. After seven irradiations, the subgroup receiving 40 mW experienced a drop in microvessel number, but it was still higher than in the control groups. CONCLUSIONS: LLLT of different intensities seems to promote neovascularization in damaged Achilles tendons of rats after partial rupture compared to controls. PMID- 16262578 TI - Morphological study of the Er,Cr:YSGG laser for root canal preparation in mandibular incisors with curved root canals. AB - OBJECTIVE: The objectives of this study were to observe morphological changes of root canal walls and to evaluate the capability of Er,Cr:YSGG laser for preparation of curved mandibular incisor root canals in vitro. BACKGROUND DATA: There are no published reports on the use of Er,Cr:YSGG laser for curved root canal preparation. METHODS: Twenty curved mandibular incisor root canals (10 and 15 degrees) were prepared sequentially by laser irradiation at 20 Hz, 2 Wusing two different sizes of fine fiber tips (200 and 320 microm in diameters). After laser irradiation, the teeth were bisected longitudinally, and the specimens were observed by stereoscope and scanning electron microscope (SEM). RESULTS: Root canal walls of the irradiated areas showed step-like appearances and openings of dentinal tubules, and root canal preparation, having curvatures of less than 10 degrees, could be carried out by this laser irradiation. CONCLUSIONS: Er,Cr:YSGG laser irradiation is capable of preparing root canals having curvatures of less than 10 degrees. PMID- 16262577 TI - In vitro effect of nitrogen and He-Ne laser on the apoptosis of human polymorphonuclear cells from burn cases and healthy volunteers. AB - OBJECTIVE: The aim of this study was to examine the effect of He-Ne and nitrogen lasers on the apoptosis of PMN in normal versus burn patients. BACKGROUND DATA: Nitrogen and He-Ne laser exposure increases the apoptotic death rate for human macrophages. Inflammation is a major consequence of thermal injury, and polymorphonuclear cell (PMN) infiltration exacerbates inflammatory process through the release of proinflammatory cytokines. The apoptotic death instead of necrotic death of PMN under the situation may help to resolve inflammation. METHODS: Ten healthy volunteers and 10 burn cases (30-50% burn surface) were included in the study. The PMN was separated by dextran sedimentation and density gradient centrifugation before suspending in RPMI-1640 medium supplemented with autologus serum. The cell suspension aliquoted in microwells was exposed to nitrogen (wavelength of 337 nm with power output of 3 mW) and He-Ne (LGN model no. 111, Russia, wavelength of 632.8 nm with power output of 3 mW) lasers for 10 and 5 min. The wells not exposed to laser were used as controls. After 24-36 h of incubation, the apoptotic rates were measured as percentage by morphological studies on acridine orange-ethidium bromide stained preparation using fluorescent microscope. RESULTS: Percentage of apoptotic death increases from 32.9% (SD +/- 4.14) in control PMN to 41.97% (SD +/- 14) in PMN exposed to nitrogen laser for 5 min and further increased to 62.7% (SD +/- 15.11) with nitrogen laser exposure for 10 min. He-Ne laser exposure for 10 min increased apoptotic cell percentage to 41.9%. Increased apoptosis in PMN exposed to nitrogen laser was statistically significant (p < 0.03) both for PMN from healthy subjects and burn cases. It was significantly elevated (p = 0.005) only for PMN from healthy volunteers exposed to He-Ne laser for 10 min but not among He-Ne exposed PMN from burn cases. CONCLUSIONS: These observations support the therapeutic application of nitrogen laser to reduce inflammation and improve wound healing for burn cases. PMID- 16262579 TI - Polarized light (400-2000 nm) and non-ablative laser (685 nm): a description of the wound healing process using immunohistochemical analysis. AB - OBJECTIVE: This study aimed to describe, through morphologic and cytochemical analysis, the healing process of wounds submitted (or not) to laser therapy (lambda685 nm) or polarized light (lambda400-2000 nm). BACKGROUND DATA: There are many reports on different effects of several types of phototherapies on the treatment of distinct conditions, amongst them, on wound healing. Laser therapy and the use of polarized light are still controversial despite successive reports on their positive effects on several biological processes. METHODS: Thirty male Wistar rats, approximately 4 months old, were used, and standardized excisional wounds were created on their dorsum. The wounds were irradiated in four equidistant points with laser light or illuminated with polarized light, both with doses of 20 or 40 J/cm2. Group 1 acted as untreated controls. Animals were irradiated every 48 h during 7 days, starting immediately after surgery, and were humanely killed on the 8th post-operative day. Specimens were taken and routinely processed and stained with H&E, and for descriptive analysis of myofibroblasts and collagen fibers, the specimens were imunnomarked by smooth muscle alpha-actin and picrosirius stain. RESULTS: Control specimens showed the presence of ulceration, hyperemia, discrete edema, intense, and diffuse inflammation, collagen deposition was irregular, and myofibroblasts were seen parallel to the wound margins. Wounds treated by laser therapy with a dose of 20 J/cm2 showed mild hyperemia, inflammation varied from moderate to intense, the number of fibroblasts was large, and the distribution of collagen fibers was more regular. Increasing the dose to 40 J/cm2 evidenced exuberant neovascularization, severe hyperemia, moderate to severe inflammation, large collagen deposition, and fewer myofibroblasts. On subjects illuminated with polarized light with a dose of 20 J/cm2, mild to moderate hyperemia was detectable, and collagen matrix was expressive and unevenly distributed; a larger number of myofibroblasts was present and no re-epithelialization was seen. Increasing the dose resulted in mild to moderate hyperemia, no re-epithelialization was seen, edema was discrete, and inflammation was moderate. CONCLUSION: The use of 685-nm laser light or polarized light with a dose of 20 J/cm2 resulted in increased collagen deposition and better organization on healing wounds, and the number of myofibroblast was increased when polarized light is used. PMID- 16262580 TI - Bond strength of a dentin bonding system using two techniques of polymerization: visible-light and argon laser. AB - OBJECTIVE: The aim of this work was to study one dentin-bonding system associated with posterior teeth restorative composite resin by means of tensile bond strength tests varying the technique of polymerization: visible light and argon laser. BACKGROUND DATA: Previous studies have demonstrated the ability of the argon laser to polymerize light-activated materials. METHODS: Sixty specimens were prepared by grinding the labial surface of bovine teeth embedded in acrylic resin. The dentin bonding system used was Single Bond (3M), which has a poliacenoic acid copolimer, associated with a posterior teeth restorative composite resin (Filtek P60, 3M). The bonding sites were treated according to the instruction of the manufacturers. The 60 teeth, duly embedded and ground, were assigned to four groups with 15 teeth each: group 1, the adhesive was light cured during 10 sec with visible light (Curing Light, 3M) with power density of 410 mW/cm2 and the composite resin was light cured during 20 sec with visible light; group 2, the adhesive and the composite resin were cured during 10 seconds with argon laser with 150 mW of power; group 3, the adhesive and the composite resin were cured during 10 sec with argon laser with 200 mW of power; and group 4, the adhesive and the composite resin were cured during 10 sec with argon laser with 250 mW of power. The composite resin was light cured in layers of 1 mm of thickness until the model of teflon with 3 mm in height was completely filled. RESULTS: The tensile bond strength test was performed in a Mini-Instron (model 4442) and the results for group 1 were 19.75 MPa (+/-4.65), group 2 were 16.09 MPa (+/-7.27), group 3 were 11.56 MPa (+/-4.50), and group 4 were 11.90 MPa (+/ 5.78). CONCLUSIONS: One can conclude that the tensile bond strength promoted by the polymerization with visible light presented greater tensile bond strength than the polymerization with argon laser with 200 mW and 250 mW, but there was no significant difference between visible light and argon laser with 150 mW. There was no significant difference between argon laser with 150 mW and argon laser with 200 mW or 250 mW. PMID- 16262581 TI - Detection of incipient carious lesions formed on human teeth in vitro using ultraviolet laser. AB - OBJECTIVE: A 325-nm ultraviolet (UV) laser was tested for effectiveness in the detection of incipient carious lesions on teeth. BACKGROUND DATA: The combination of early detection with new intervention methods and caries management will be the preferred dentistry of the future. METHODS: Carious lesions from extracted teeth or from demineralized teeth were irradiated by UV laser, and the fluorescence spectrum was measured. The peak ratio between two peak intensities in the spectrum was determined. RESULTS: Fluorescence peak intensity of approximately 425 nm was gradually decreased as carious lesions formed, whereas the change 625 nm was less significant. The peak ratio change between peak intensities of approximately 425 and at 625 nm was observed after 30 min of demineralization. Morphological changes were barely observable in this range. CONCLUSIONS: A 325-nm UV laser showed high efficacy in the detection of incipient carious lesions. By evaluating peak ratio, the incipient carious lesions could be detected. PMID- 16262582 TI - Effects of Er:YAG and Nd:YAG lasers on dentin permeability in root surfaces: a preliminary in vitro study. AB - OBJECTIVE: This in vitro study evaluated the effects of Nd:YAG and Er:YAG lasers on reducing dentin permeability by sealing opened tubules. BACKGROUND DATA: According to hydrodynamic theory, dentine hypersensitivity occurs when dentin is exposed with tubules opened. Consequently, a painful sensation occurs due to an intensification of the dentinal permeability. Treatment, therefore, should be based on a decrease of this permeability, achieved by the obliteration of dentinal tubules. The Nd:YAG laser is known for its capacity to seal dentinal tubules; however, few studies concerning treatment with Er:YAG laser are available. METHODS: The Nd:YAG laser was used, based on two parameters: (A) 1.0 W, 10 Hz, and (B) 1.5 W, 15 Hz. The Er:YAG laser was used at 60 mJ, 2 Hz, four applications of 20 sec each, at 6 mm from the surface. After irradiation, all samples were immersed in 1% Rodamine B dye solution, in order to evaluate the penetration of the dye solution and observe the decrease/increase of dentinal permeability after the laser treatment. RESULTS: The laser conditions used in the present study decreased the permeability as follows: (a) when using the Er:YAG laser at 60 mJ, 2 Hz in 26.05%, and (b) when using the Nd:YAG laser at 1.5 W, 15 Hz in 19.03%--with no statistical difference between them. Additionally, the Nd:YAG laser at 1.0 W, 10 Hz, decreased permeability in 4.59%, with a smaller effect. CONCLUSIONS: According to the results of this in vitro study, the Er:YAG laser at 60 mJ, 2 Hz, and the Nd:YAG laser at 1.5 W, 15 Hz are useful for decreasing dentin permeability. PMID- 16262584 TI - Laser literature watch. PMID- 16262583 TI - Combining the CO2 laser and the endoscope to remove soft tissue masses from the forehead area. AB - OBJECTIVE: The aim of this study was to provide a retrospective analysis of treatment that combines the laser and the endoscope. BACKGROUND DATA: Previously, surgeons faced many complications arising from endoscope-assisted surgery in the region of the forehead. In order to reduce the incidence of complications, such as swelling and hematoma, the CO2 laser is added to endoscope-assisted surgery. METHODS: Between January 1996 and January 2003, 42 patients were treated for the removal of a benign tumor. The group was comprised of 18 males and 24 females, and their ages ranged from 5 to 64 years. The SurgiLase 150XJ CO2 laser system, used in conjunction with the flexible FIBERLASE (Sharplan, NJ), was used in all cases. The endoscope was 4 mm in diameter, with an angle of 30 degrees (Snoden Pancer, USA). The dissection was performed by relaying images from the endoscope to a monitor. The basic instrumentation consists of elevators, a nerve hook, retractors, and endoscopic scissors. Suction of the smoke generated by laser evaporization was also necessary. RESULTS: Of the 42 cases treated, 30 patients displayed lipomas, 10 patients displayed inclusion cysts, and two patients displayed dermoid cysts. After surgery, the follow-up period ranged from 6 months, to 2 years and 6 months, with an average of 1 year and 7 months. In this retrospective review, the incidences of early complications of these 42 patients included one case of scalp alopecia, three of scar alopecia, one of numbness, and three of ecchymosis. No hematoma cases were recorded. Evaluation after 6 months revealed the later complication of one scar alopecia. CONCLUSIONS: SurgiLase CO2 laser, used in conjunction with endoscopic surgery, enables plastic surgeons to work at a distance through small incisions without causing bleeding. Visible scars are reduced and recovery time is diminished. PMID- 16262586 TI - Recent pharmacologic updates. PMID- 16262587 TI - Consumer driven health care. PMID- 16262588 TI - The need to study substance use disorders. PMID- 16262589 TI - Naltrexone treatment of adolescent alcoholics: an open-label pilot study. AB - OBJECTIVE: This 6-week open-label trial of naltrexone was conducted in a preliminary fashion to determine whether naltrexone would be safe, well tolerated, and lead to a reduction in alcohol consumption in adolescents with alcohol dependence. METHOD: Five outpatient treatment-seeking adolescents who met Diagnostic and Statistical Manual of Mental Disorders, 4th edition (DSM-IV) criteria for alcohol dependence were recruited. The Child Schedule for Schizophrenia and Affective Disorders (K-SADS), Structured Clinical Interview for DSM (SCID), and the Family History Questionnaire were administered at baseline. The Time-Line Follow-Back (TLFB) and two craving scales (Adolescent Obsessive Compulsive Drinking Scale [A-OCDS] and a craving analog scale) were administered at baseline and weekly thereafter. Each subject received a 10-day supply of naltrexone (50 mg) and a 100-mg riboflavin capsule. Subjects were instructed to take naltrexone and riboflavin simultaneously. RESULTS: Overall, the average drinks per drinking day (DDD) decreased significantly from baseline to the end of week 6 with an average reduction of 7.61 standard drinks. There was a significant reduction in the average A-OCDS total score, A-OCDS Irresistibility subscale score, and craving analog score. Nausea was the only side-effect reported, and there were no elevations of liver enzymes. Naltrexone was well tolerated by the alcohol-dependent adolescent. CONCLUSIONS: Our data suggest that naltrexone is safe and well tolerated in adolescent alcoholics. Naltrexone may lead to a significant reduction in alcohol consumption and craving in adolescent alcoholics, but larger, randomized, controlled trials are needed. PMID- 16262590 TI - A pilot study of methylphenidate preference assessment in children diagnosed with attention-deficit/hyperactivity disorder. AB - OBJECTIVE: The use of methylphenidate (MPH) in the treatment of attention deficit/hyperactivity disorder (ADHD) is widely accepted; however, there is increased concern regarding its abuse potential. Few studies have examined the reinforcing effects of drugs in individuals receiving them for clinical purposes. This study attempts to assess MPH preference in children with ADHD using a choice procedure in order to explore the relationship among drug preference, clinical efficacy, and abuse potential. METHODS: Participants were 5 children (10-14 years of age) receiving MPH for the treatment of ADHD. Reinforcing effects were assessed using a double-blind choice procedure, with six sampling sessions and six choice sessions. Participant-rated effects were measured using self-report questionnaires. Clinical effects were measured using direct observations and behavior ratings. RESULTS: Differences between the number of MPH, Placebo, and Neither choices across participants were significant (chi2 = 9.6; p < 0.01). Three of five participants reliably chose MPH more often than placebo. MPH produced idiosyncratic patterns of participant-rated effects but failed to produce significant clinical effects. CONCLUSIONS: These findings add to the literature on the reinforcing effects of MPH and are the first reported in a clinical sample of children. Further research exploring the role of clinical efficacy in MPH preference is warranted. PMID- 16262591 TI - Assessing cannabis use in adolescents and young adults: what do urine screen and parental report tell you? AB - OBJECTIVES: Our analysis compares three approaches to detect the most common drug abused in early adulthood, cannabis: (1) report on direct structured interview; (2) indirect parental report; and (3) urine toxicology screen. METHODS: We examined data on 207 subjects (36% also met criteria for alcohol abuse; 9% for alcohol dependence) derived from two prospective and ongoing family studies of boys and girls with or without attention-deficit/hyperactivity disorder (ADHD). Assessments relied on the Schedule for Affective Disorders and Schizophrenia (K SADS-E; under 18 years of age) and on the Structured Clinical Interview for DSM IV (SCID-IV; over 18 years of age). Urine samples were analyzed with Auccusign DOA5 (on-site screening assay). RESULTS: Ninety-seven percent (97%) of individuals, who reported no use of cannabis within the past month, had a negative urine screening and 79% of individuals, who endorsed cannabis abuse/dependence, had a positive urine screening. The sensitivity of the direct structured interview report was 91%, the specificity 87%, the positive predicting value 67%, and the negative predictive value 97%. Indirect parental reports were found to be less informative on cannabis use than direct report. CONCLUSION: Direct report of cannabis use, abuse, or dependence during the structured interview is both sensitive and specific when compared to urine toxicology screens and indirect parental reports. PMID- 16262592 TI - Attention-deficit/hyperactivity disorder and early-onset substance use disorders. AB - In recent years, there has been an increased recognition of the common comorbidity of attention-deficit/ hyperactivity disorder (ADHD) and substance use disorder (SUD) among adolescents and adults. ADHD can be an important factor in the pathogenesis and maintenance of SUD; moreover, retrospective studies suggest that treating ADHD during childhood may prevent the development of SUD. In addition, treatment of ADHD among adults, and possibly adolescents, with SUD can reduce their risk of relapse. Theoretical mechanisms that may explain the relationship between ADHD and SUD are explored in this paper. Current research and recommended clinical practices related to the diagnosis and treatment of ADHD with SUD in adolescents are discussed as well. More research is needed to definitively assess the effectiveness and safety of medications in this population of youths with ADHD and SUD. PMID- 16262593 TI - Psychostimulant treatment and risk for substance abuse among young adults with a history of attention-deficit/hyperactivity disorder: a population-based, birth cohort study. AB - OBJECTIVE: The aim of this study was to evaluate the association between stimulant treatment and the risk for substance abuse among young adults with a childhood diagnosis of attention- deficit/hyperactivity disorder (ADHD). METHODS: Subjects included 295 research-identified ADHD incidence cases treated with psychostimulant medication and 84 ADHD cases not treated with psychostimulants. These subjects are from a 1976-1982 population-based birth cohort, retrospectively, followed from birth until emigration, death, or last follow-up (mean = 17.2 years of follow-up). Medical and school records were reviewed for documented substance abuse and psychostimulant treatment. The association was evaluated using logistic regression models. RESULTS: Socioeconomic characteristics at birth, and comorbidities, were similar between treated and untreated ADHD cases. Sixty (20.3%) of treated ADHD cases had documented substance abuse compared to 23 (27.4%) of cases not treated (OR = 0.7; 95% CI = 0.4-1.2). Among treated ADHD boys, 21.8% had substance abuse compared to 36.4% not-treated ADHD boys (OR = 0.5; 95% CI = 0.3-0.9). Among treated ADHD girls, 15.2% had substance abuse compared to 10.3% not-treated ADHD girls (OR = 1.5; 95% CI = 0.4-6.1). CONCLUSION: While these results cannot demonstrate cause and effect, our findings indicate that psychostimulant treatment of childhood ADHD is associated with reduced risk for later substance abuse among boys with ADHD. PMID- 16262594 TI - Bupropion SR for the treatment of substance-abusing outpatient adolescents with attention-deficit/hyperactivity disorder and mood disorders. AB - OBJECTIVE: Few studies exist on pharmacological interventions for adolescents with substance use disorders (SUD). To this end, we evaluated the response of bupropion hydrochloride sustained release (SR) in SUD adolescents with comorbid psychopathology (both attention-deficit/ hyperactivity disorder (ADHD) and a mood disorder). METHODS: Fourteen adolescent outpatients were treated naturalistically and followed openly for 6 months. Adolescents were rated using the Drug Use Screening Inventory--Revised (DUSI-R), ADHD Symptom Checklist, and the Hamilton Rating Scale for Depression (HAM-D). Clinical Global Impression (CGI) Scale scores were obtained for Substance Abuse, ADHD, Anxiety, and Depression. The ratings were completed at baseline, at month 3, and at the 6-month endpoint. Bupropion SR was initiated at 100 mg once-daily and titrated naturalistically to a maximum dose of 400 mg/day. RESULTS: Of the 14 subjects followed, 13 subjects completed 6 months of treatment. At the 6- month endpoint compared to baseline, treatment with bupropion was associated with clinical and significant reductions in DUSI scores (-39%; p < 0.05), ADHD symptom checklist (-43%; p < 0.001), HAM-D (-76%; p < or = 0.001); and reductions in the CGIs for ADHD (p < or = 0.001), depression (p < or = 0.001), and substance abuse (p < 0.05). The mean daily dose of bupropion SR was 315 mg (in divided doses). No significant adverse events were noted during the follow-up period. CONCLUSIONS: These naturalistic data suggest that bupropion is well tolerated and may be an effective medication for the treatment of substance abusing adolescents with comorbid mood disorders and ADHD. PMID- 16262595 TI - The clinical dilemma of using medications in substance-abusing adolescents and adults with attention-deficit/hyperactivity disorder: what does the literature tell us? AB - OBJECTIVE: Adolescents and adults with attention-deficit/hyperactivity disorder (ADHD) and substance use disorders (SUD) are increasingly recognized in clinical practice. The role of pharmacological treatment for ADHD in these comorbid individuals remains unclear. METHODS: A systematic review of the medical literature was conducted through PubMed, supplemented with data from scientific presentations, to evaluate the role of medication treatment of ADHD in substance abusing individuals with ADHD. Meta-analysis was used to evaluate the effects of medication therapy on ADHD and SUD outcomes in general, while specifically addressing trial design, trial duration, retention, class of medication, age group, concurrent psychotherapy, and outcome in both SUD and ADHD domains. RESULTS: Four studies in adolescents and five studies in adults with ADHD plus SUD were identified (two controlled and seven open studies; n = 222 subjects). The standard mean difference (SMD) indicated statistically significant improvements in ADHD and SUD that were not maintained when evaluating controlled studies only. Albeit limited by power, trial duration, retention rate, and age group did not influence outcome. No worsening of SUD or drug-drug interactions were observed in any of the studies. The results could not be accounted for by any single study or by publication bias. CONCLUSION: Treating ADHD pharmacologically in individuals with ADHD plus SUD has a moderate impact on ADHD and SUD that is not observed in controlled trials and does not result in worsening of SUD or adverse interactions specific to SUD. Further controlled trials evaluating the effect of novel combinations of psychotherapy and ADHD pharmacotherapy on SUD relapse in these groups are warranted. PMID- 16262596 TI - Attention-deficit/hyperactivity disorder, medication treatment, and substance use patterns among adolescents and young adults. AB - OBJECTIVE: The aim of this study was to examine the relationship between current active attention-deficit/hyperactivity disorder (ADHD) symptoms, medication treatment, and substance use patterns among college students. METHOD: Three hundred and thirty-four students at a local college were surveyed for current ADHD symptoms and psychopharmacological treatment. The survey was conducted in conjunction with an annual national survey that probes students about their substance use patterns and attitudes. RESULTS: Participants with ADHD as ascertained by medication treatment of ADHD had greater past-year tobacco and marijuana use. Among those with ADHD, participants with active ADHD symptoms were more likely to have past-year tobacco and other drug (besides tobacco, alcohol, and marijuana) use as compared to those without active ADHD symptoms. In addition, participants with active ADHD symptoms were more likely to have past month "other" drug use as compared to those without active ADHD symptoms. Among those prescribed medications for ADHD, 25% reported ever using their medication to "get high" and almost 29% reported ever giving or selling their medication to someone else. CONCLUSIONS: Results of our preliminary study indicated that ADHD symptom control may be important to protect against increased risk of substance use (particularly tobacco and drugs other than alcohol and marijuana) among college-age students with ADHD. Further studies of misuse/diversion of prescription stimulant medication among college students are needed. PMID- 16262597 TI - Pharmacotherapy of adolescent substance use disorders: a review of the literature. AB - OBJECTIVE: Substance use disorders (SUD) are increasingly conceptualized as developing during adolescence. Moreover, adolescent SUD are viewed as one of the most difficult mental health conditions to treat. The role of pharmacotherapy for adolescents with SUD is not well delineated. We systematically reviewed existing pharmacotherapy studies of adolescent SUD. METHODS: A computerized search of the literature addressing the pharmacotherapy of SUD in adolescents was performed. Data from relevant peer-reviewed scientific presentations were also included. RESULTS: Five case reports (n = 8 subjects), six open studies that primarily addressed SUD with comorbid psychiatric disorders (n = 73 subjects), and five placebo-controlled studies (n = 156 subjects) on the pharmacological treatment of youths with SUD were identified. Pharmacological agents appear to reduce comorbid psychopathology with a milder reduction in SUD symptoms. The most robust evidence exists for the treatment of SUD comorbid with affective disorders. There is a limited database supporting the use of agents to reduce substance craving and the preventative effects of pharmacotherapy on subsequent SUD development in adolescents with specific psychiatric disorders. CONCLUSION: Further controlled studies directed at reducing drug use and craving, and guiding the integration of pharmacotherapy and psychosocial interventions for SUD with the treatment of comorbid psychiatric illness, are necessary. PMID- 16262598 TI - Integrated substance use and mental health treatment for adolescents: aligning organizational and financial incentives. AB - The high prevalence of the dual diagnosis of mental and substance use disorders (SUD) has been increasingly documented for both adolescents and adults (Crowley and Riggs 1995; Kandel et al. 1999; Whitmore et al. 1997). For more than a decade, the National Institute of Drug Abuse (NIDA) has included integrated treatment of comorbid psychiatric disorders as one of nine core treatment principles (National Institute on Drug Abuse 1999). Despite empirically supported practice guidelines, implementation of integrated treatment has been slow (New Freedom Commission on Mental Health 2003; U.S. Department of Health and Human Services 1999). In response to the growing call for integrated treatments and systems of care, this paper: (1) identifies systemic and economic barriers that have impeded widespread implementation of integrated care for adolescents with co occurring SUD, specifically the supply of treatment providers, shifting priorities of gatekeepers to specialty care, and financing streams; and (2) describes possibilities for aligning economic incentives in order to facilitate the dissemination and implementation of integrated care for adolescents with co occurring SUD. PMID- 16262600 TI - Localization of a portion of the liver isoform of fatty-acid-binding protein (L FABP) to peroxisomes. AB - The liver isoform of fatty-acid-binding protein (L-FABP) facilitates the cellular uptake, transport and metabolism of fatty acids and is also involved in the regulation of gene expressions and cell differentiation. Consistent with these functions, L-FABP is predominantly present in the cytoplasm and to a lesser extent in the nucleus; however, a significant portion of this protein has also been detected in fractions containing different organelles. More recent observations, notably on L-FABP-deficient mice, indicated a possible direct involvement of L-FABP in the peroxisomal oxidation of long-chain fatty acids. In order to clarify the links between L-FABP and peroxisomal lipid metabolism, we reinvestigated the subcellular distribution of the protein. Analytical subcellular fractionation by a method preserving the intactness of isolated peroxisomes, two-dimensional gel electrophoresis of peroxisomal matrix proteins combined with MS analysis, and immunoelectron microscopy of liver sections demonstrate the presence of L-FABP in the matrix of peroxisomes as a soluble protein. Peroxisomal L-FABP was highly inducible by clofibrate. The induction of L-FABP was accompanied by a marked increase in the binding capacity of peroxisomal matrix proteins for oleic acid and cis-parinaric acid. The peroxisomal beta-oxidation of palmitoyl-CoA and acyl-CoA thioesterase activity were stimulated by L-FABP, indicating that the protein modulates the function of peroxisomal lipid-metabolizing enzymes. The possible role of intraperoxisomal L FABP in lipid metabolism is discussed. PMID- 16262601 TI - Identification of the anti-inflammatory protein tristetraprolin as a hyperphosphorylated protein by mass spectrometry and site-directed mutagenesis. AB - Tristetraprolin (TTP) is a zinc-finger protein that binds to AREs (AU-rich elements) within certain mRNAs and causes destabilization of those mRNAs. Mice deficient in TTP develop a profound inflammatory syndrome with erosive arthritis, autoimmunity and myeloid hyperplasia. Previous studies showed that TTP is phosphorylated extensively in intact cells. However, limited information is available about the identities of these phosphorylation sites. We investigated the phosphorylation sites in human TTP from transfected HEK-293 cells by MS and site-directed mutagenesis. A number of phosphorylation sites including Ser66, Ser88, Thr92, Ser169, Ser186, Ser197, Ser218, Ser228, Ser276 and Ser296 were identified by MS analyses using MALDI (matrix-assisted laser-desorption ionization)-MS, MALDI-tandem MS, LC (liquid chromatography)-tandem MS and multidimensional protein identification technology. Mutations of Ser197, Ser218 and Ser228 to alanine in the human protein significantly increased TTP's gel mobility (likely to be stoichiometric), whereas mutations at the other sites had little effect on its gel mobility. Dephosphorylation and in vivo labelling studies showed that mutant proteins containing multiple mutations were still phosphorylated, and all were able to bind to RNA probes containing AREs. Confocal microscopy showed a similar cytosolic localization of TTP among the various proteins. Ser197, Ser218 and Ser228 are predicted by motif scanning to be potential sites for protein kinase A, glycogen synthase kinase-3 and extracellular-signal-regulated kinase 1 (both Ser218 and Ser228) respectively. The present study has identified multiple phosphorylation sites in the anti inflammatory protein TTP in mammalian cells and should provide the molecular basis for further studies on the function and regulation of TTP in controlling pro-inflammatory cytokines. PMID- 16262602 TI - Regulation of cysteine dioxygenase degradation is mediated by intracellular cysteine levels and the ubiquitin-26 S proteasome system in the living rat. AB - Mammalian metabolism of ingested cysteine is conducted principally within the liver. The liver tightly regulates its intracellular cysteine pool to keep levels high enough to meet the many catabolic and anabolic pathways for which cysteine is needed, but low enough to prevent toxicity. One of the enzymes the liver uses to regulate cysteine levels is CDO (cysteine dioxygenase). Catalysing the irreversible oxidation of cysteine, CDO protein is up-regulated in the liver in response to the dietary intake of cysteine. In the present study, we have evaluated the contribution of the ubiquitin-26 S proteasome pathway to the diet induced changes in CDO half-life. In the living rat, inhibition of the proteasome with PS1 (proteasome inhibitor 1) dramatically stabilized CDO in the liver under dietary conditions that normally favour its degradation. Ubiquitinated CDO intermediates were also seen to accumulate in the liver. Metabolic analyses showed that PS1 had a significant effect on sulphoxidation flux secondary to the stabilization of CDO but no significant effect on the intracellular cysteine pool. Finally, by a combination of in vitro hepatocyte culture and in vivo whole animal studies, we were able to attribute the changes in CDO stability specifically to cysteine rather than the metabolite 2-mercaptoethylamine (cysteamine). The present study represents the first demonstration of regulated ubiquitination and degradation of a protein in a living mammal, inhibition of which had dramatic effects on cysteine catabolism. PMID- 16262603 TI - Induction of spermidine/spermine N1-acetyltransferase (SSAT) by aspirin in Caco-2 colon cancer cells. AB - Epidemiological, experimental and clinical results suggest that aspirin and other NSAIDs (non-steroidal anti-inflammatory drugs) inhibit the development of colon cancer. It has been shown that the NSAID sulindac induces apoptosis and suppresses carcinogenesis, in part, by a mechanism leading to the transcriptional activation of the gene encoding SSAT (spermidine/spermine N1-acetyltransferase), a rate-limiting enzyme in polyamine catabolism. In the present study, we show that a variety of NSAIDs, including aspirin, sulindac, ibuprofen and indomethacin, can induce SSAT gene expression in Caco-2 cells. Aspirin, at physiological concentrations, can induce SSAT mRNA via transcriptional initiation mechanisms. This induction leads to increased SSAT protein levels and enzyme activity. Promoter deletion analysis of the 5' SSAT promoter-flanking region led to the identification of two NF-kappaB (nuclear factor kappaB) response elements. Electrophoretic mobility-shift assays showed binding of NF-kappaB complexes at these sequences after aspirin treatment. Aspirin treatment led to the activation of NF-kappaB signalling and increased binding at these NF-kappaB sites in the SSAT promoter, hence providing a potential mechanism for the induction of SSAT by aspirin in these cells. Aspirin-induced SSAT ultimately leads to a decrease in cellular polyamine content, which has been associated with decreased carcinogenesis. These results suggest that activation of SSAT by aspirin and different NSAIDs may be a common property of NSAIDs that plays an important role in their chemopreventive actions in colorectal cancer. PMID- 16262606 TI - Strengthening the prevention of oral cancer: the WHO perspective. PMID- 16262604 TI - Domain II plays a crucial role in the function of ribosome recycling factor. AB - RRF (ribosome recycling factor) consists of two domains, and in concert with EF-G (elongation factor-G), triggers dissociation of the post-termination ribosomal complex. However, the function of the individual domains of RRF remains unclear. To clarify this, two RRF chimaeras, EcoDI/TteDII and TteDI/EcoDII, were created by domain swaps between the proteins from Escherichia coli and Thermoanaerobacter tengcongensis. The ribosome recycling activity of the RRF chimaeras was compared with their wild-type RRFs by using in vivo and in vitro activity assays. Like wild-type TteRRF (T. tengcongensis RRF), the EcoDI/TteDII chimaera is non functional in E. coli, but both wild-type TteRRF, and EcoDI/TteDII can be activated by coexpression of T. tengcongensis EF-G in E. coli. By contrast, like wild-type E. coli RRF (EcoRRF), TteDI/EcoDII is fully functional in E. coli. These findings suggest that domain II of RRF plays a crucial role in the concerted action of RRF and EF-G for the post-termination complex disassembly, and the specific interaction between RRF and EF-G on ribosomes mainly depends on the interaction between domain II of RRF and EF-G. This study provides direct genetic and biochemical evidence for the function of the individual domains of RRF. PMID- 16262605 TI - Increased plasma angiotensin II in postural tachycardia syndrome (POTS) is related to reduced blood flow and blood volume. AB - POTS (postural tachycardia syndrome) is associated with low blood volume and reduced renin and aldosterone; however, the role of Ang (angiotensin) II has not been investigated. Previous studies have suggested that a subset of POTS patients with increased vasoconstriction related to decreased bioavailable NO (nitric oxide) have decreased blood volume. Ang II reduces bioavailable NO and is integral to the renin-Ang system. Thus, in the present study, we investigated the relationship between blood volume, Ang II, renin, aldosterone and peripheral blood flow in POTS patients. POTS was diagnosed by 70 degrees upright tilt, and supine calf blood flow, measured by venous occlusion plethysmography, was used to subgroup POTS patients. A total of 23 POTS patients were partitioned; ten with low blood flow, eight with normal flow and five with high flow. There were ten healthy volunteers. Blood volume was measured by dye dilution. All biochemical measurements were performed whilst supine. Blood volume was decreased in low-flow POTS (2.14 +/- 0.12 litres/m2) compared with controls (2.76 +/- 0.20 litres/m2), but not in the other subgroups. PRA (plasma renin activity) was decreased in low flow POTS compared with controls (0.49 +/- 0.12 compared with 0.90 +/- 0.18 ng of Ang I.ml(-1).h(-1) respectively), whereas plasma Ang II was increased (89 +/- 20 compared with 32 +/- 4 ng/l), but not in the other subgroups. PRA correlated with aldosterone (r = +0.71) in all subjects. PRA correlated negatively with blood volume (r = -0.72) in normal- and high-flow POTS, but positively (r = +0.65) in low-flow POTS. PRA correlated positively with Ang II (r = +0.76) in normal- and high-flow POTS, but negatively (r = -0.83) in low-flow POTS. Blood volume was negatively correlated with Ang II (r = -0.66) in normal- and high-flow POTS and in five low-flow POTS patients. The remaining five low-flow POTS patients had reduced blood volume and increased Ang II which was not correlated with blood volume. The data suggest that plasma Ang II is increased in low-flow POTS patients with hypovolaemia, which may contribute to local blood flow dysregulation and reduced NO bioavailability. PMID- 16262607 TI - For debate: problems with the DMF index pertinent to dental caries data analysis. AB - The Decayed, Missing, Filled (DMF) index has been used for over 50 years and is well established as the key measure of caries experience in dental epidemiology. Despite its long history of use, there is debate about the most appropriate number of surfaces to include for a missing tooth. Assigning the maximum possible value for the 'M' component of DMFS (Surfaces) leads to overestimation of an individual's caries experience, and in any associated comparisons of in-caries experience, whereas assigning the minimum possible value for the 'M' component has the opposite effect. Alternative methods of assigning the number of caries affected surfaces for an extracted tooth are considered. The net caries increment and adjusted caries increment (common methods of correction of the crude increment measure for reversals) are discussed, along with incidence density, a measure of caries extent. Problems exist with the adjusted caries increment, particularly among cohorts with low mean baseline caries experience. Development of an alternative method of estimating the relationship of 'true' and 'examiner' reversals is advocated, as well as greater utilization of incidence density in dental epidemiology. PMID- 16262608 TI - Exposure to 'ideal' facial images reduces facial satisfaction: an experimental study. AB - OBJECTIVE: Psychological research has suggested that exposure to media images of 'ideal' female models has a negative impact on women's perception of their body image. The present study examined the effects of exposure to images of faces rated as high in attractiveness upon facial satisfaction, compared with exposure to 'neutral' stimuli. METHODS: Participants comprised three groups of women: 24 women undergoing orthodontic treatment, 22 patients undergoing orthognathic treatment, and 20 women receiving no current dental treatment. All completed measures of facial and body image satisfaction after viewing images of houses or faces as follows: The Body Satisfaction Scale, The Revised Body Image Scale of Secord and Jourard and a Visual Analogue Scale measuring satisfaction with Facial Appearance. A repeated-measures design was adopted with a 4-6-week period between experimental conditions. All participants were randomly assigned to one of the two orders (Houses-Faces or Faces-Houses). The findings were analysed using general linear modelling analysis of variance. RESULTS: Exposure to idealized images of faces resulted in a significant decrease in facial satisfaction when compared with the control condition (mean facial satisfaction for all participants in Faces condition, Body Satisfaction Scale-Head Scale = 22.2, SD = 7.81; Revised Body Image Scale-Facial Subscale = 25.5, SD = 4.57; VAS = 7.8, SD = 2.47; Mean facial satisfaction for all participants in Houses condition, Body Satisfaction Scale-Head scale = 19.5, SD = 6.81; Revised Body Image Scale-Facial Subscale = 26.8, SD = 4.87; VAS = 6.2, SD = 2.72). CONCLUSIONS: Media may exert a negative influence on self-perception, particularly amongst individuals who, for one reason or another, are sensitive to the appearance of their face. The findings of the present study have implications for the demand for orthodontic treatment for aesthetic enhancement at a time when resources are limited. Interventions are discussed to help individuals question their acceptance of such media messages. PMID- 16262609 TI - The theory of reasoned action and patient compliance during orthodontic treatment. AB - OBJECTIVES: The aim of the present study was to investigate the potential of the theory of reasoned action (TRA) for the prediction and understanding of patients' intention to comply during orthodontic treatment and to analyze the effect of two additional variables in the model, namely perceived behavioral control and anticipated regret. Moreover, (the determinants of) intentions of orthodontic patients to comply during treatment were compared with (the determinants of) intentions of parents to stimulate this cooperation. METHODS: A questionnaire was handed out to patients and parents visiting the Department of Orthodontics of the Academic Centre of Dentistry in Amsterdam. In both the patient and parent sample, independent-sample t-tests, correlation analyses and stepwise regression analyses were conducted. Variables in both samples were compared and tested. RESULTS: The extended version of the TRA explained 20% of the variance in the patients' intention to comply. The patients' anticipated regret, attitude and motivation to comply were significant determinants of the patients' intention to comply. In addition, the parents' attitude toward compliance was a significant predictor. The role of parents in enhancing patients' intentions to comply cannot be neglected. CONCLUSION: Our findings suggest that patients' intentions to comply during orthodontic treatment are influenced by factors outside of the TRA. Therefore, it is recommended to develop a new model, in which factors of the TRA are included, which can be used specifically for the study of compliance in orthodontics. PMID- 16262611 TI - Photographs as a means of assessing developmental defects of enamel. AB - OBJECTIVES: To evaluate the validity and reliability of photographic examinations for developmental defects of enamel (DDE) in maxillary and mandibular incisors and canines using a standardized process. METHODS: The anterior teeth of 257 children were examined 'wet', both clinically and photographically for DDE, using the modified Federation Dentaire Internationale (FDI) (DDE) Index. A series of five standardized photographs were taken for each child: a frontal view perpendicular to the four incisors; two lateral views, each showing the lateral incisors and canines on each side of the dental arch; and the superior and inferior views, retaking of the frontal view with the camera held at approximately 30 degrees above and below the horizontal plane. The photographs taken for each child were viewed as three different sets; the 'five-view' (frontal, two lateral views plus superior and inferior views), 'three-view' (frontal and two lateral views), and 'one-view' (frontal view only) slide sets. RESULTS: Using 'one view' slides, 91.7% of teeth could be examined photographically. Whereas using multiple views 99.9% of teeth could be assessed. At the subject level, agreements between clinical diagnoses (gold standard) and photographic examinations were substantial to almost perfect (k = 0.73-0.86). At the tooth level, agreement was best for incisors (k = 0.71 or higher). The intra examiner reproducibility was high for the photographic assessments at both subject and tooth levels (k = 0.71-0.95). CONCLUSIONS: Multiple-view photographic slides of 'five-view' and 'three-view' are valid and reliable for assessing DDE on the 12 anterior teeth, while a 'one-view' (frontal) was acceptable to study only the incisors. PMID- 16262610 TI - Changes in health over time in patients with symptoms allegedly caused by their dental restorative materials. AB - OBJECTIVES: In Sweden, many patients with symptoms allegedly caused by their dental materials have exchanged their restorations, but the effects of the exchange have been insufficiently investigated. Therefore, the aim of the study was to describe the change in health over time for these patients and the hypothesis was that the patients could be divided based on their symptoms and that the ability to recover differs between these groups. Furthermore, we also examined if other factors such as replacement of dental restorative materials and follow-up time had any impact on the perceived health status. METHODS: A questionnaire was sent to 614 patients who had been referred to the School of Dentistry, Umea, Sweden, with symptoms allegedly caused by dental restorative materials. The response rate was 55%. RESULTS: The risk of having any further complaints was higher for patients with complex symptoms (P = 0.03) and these patients had exchanged their restorations to a significantly larger extent than the others (P = 0.03). The remaining complaints was more frequent among men (P = 0.02). Exchange of dental restorative materials had no significant impact on the ability to recover completely. However, the patients who had exchanged their restorations completely perceived a significantly larger alleviation of their symptoms than the others (P < 0.01), although the frequency of most of the symptoms had increased. CONCLUSIONS: Patients with complex symptoms had a more unfavorable long-term prognosis concerning persistent complaints than those with localized symptoms only. Furthermore, the results indicate that the patients might experience health improvements after removal of their dental restorative materials. The reason for this improvement, however, is unclear. Further analyses regarding other possible explanations than the 'odontological/medical' are needed. PMID- 16262612 TI - Dental care and HIV-infected individuals: are they equally treated? AB - OBJECTIVE: To investigate the problems in seeking dental care faced by HIV positive individuals in Italy. METHODS: A multicenter observational study was performed by distributing an anonymous self-administered questionnaire to patients of six public healthcare facilities specialized in the treatment of individuals with HIV infection. The questions concerned personal data potentially correlated with discrimination, the patient-dentist relationship before and after HIV diagnosis, and the reasons for seeking dental care in public facilities. We also evaluated the patients' discomfort in the patient-dentist relationship after HIV diagnosis, performing univariate and multivariate analyses. RESULTS: Of the 1,500 questionnaires distributed; 883 were filled-out completely. A total of 630 persons received dental care after HIV diagnosis: 209 (33.2%) did not tell the dentist that they were seropositive. Of those who did, 56 were refused care. For patients treated by a private dentist, having been treated by the same dentist before diagnosis was a risk factor for great discomfort in the patient-dentist relationship (P < 0.002). Being treated in public facilities was associated with having received dental care after HIV diagnosis (P < 0.001) and a primary school education (P < 0.001). CONCLUSIONS: There exist episodes of discrimination on the part of some dentists, and a relatively high proportion of HIV-positive persons do not disclose their seropositivity to the dentist. Dentists should be provided with training for promoting both ethically acceptable practices and suitable clinical management of HIV-positive persons. PMID- 16262613 TI - Examining the association between parenting stress and the development of early childhood caries. AB - OBJECTIVES: Numerous studies have explored the effects of pediatric health on parenting stress, but very little work has been carried out to explore the opposite and equally as compelling relationship of how parenting stress might affect child health, especially as it relates to oral health. This study examined the association between parenting stress and early childhood caries (ECC) in 97, 4- to 5-year-old Australian children attending preschools in the North Brisbane Health Region, Australia. METHODS: Using a cross-sectional study design, clinical examinations were conducted to evaluate the caries status of each child. Two caregiver questionnaires were completed -- one soliciting demographic and oral health behavior information, and the other, information on parenting stress. RESULTS: This study demonstrated a significant bivariate association between parenting stress and ECC experience as measured by dmft; however, the association did not persist in the two-part forward-selection logistic and linear regression models. A negative association between social desirability (defensive responding) and extent of caries was also determined. CONCLUSIONS: Our findings suggest the need to conduct longitudinal studies to give proper consideration to the temporal aspect of caries development and clarify the results obtained by on the relationship between parenting stress and oral health. Further study is also warranted to more clearly elucidate the association between social desirability (defensive responding) in parents and their children's ECC experience. PMID- 16262614 TI - Influence of the patient's race on the dentist's decision to extract or retain a decayed tooth. AB - OBJECTIVES: The aim of this study was to investigate the influence of the patient's race on the dentist's decision to extract or retain a decayed tooth. METHODS: A probabilistic random sample of 297 dentists from Recife, Brazil, was used. Two case scenarios were presented to the dentists. Both scenarios showed a molar that was extensively decayed, but indicated for conservative treatment. The scenarios included a description of the patient and eight photographs of the clinical case, including a photograph of the patient's face. The dentists were asked to regard the patient as poor and then to decide whether to extract or retain the molar. However, although the scenarios were based on the same clinical case, the photographs of the patient's face were different. One scenario showed the photograph of a white patient whereas the other showed the photograph of a black patient. The first scenario was presented 2 months before the second so that the dentists would not remember the former. RESULTS: The dentist's decision varied significantly according to the patient's race, with dentists deciding to extract more frequently for the black patient than for the white patient (25.6% vs. 16.2%; P < 0.001). This racial variation occurred regardless of the demographic and socioeconomic variables of the dentists. It did, however, occur as a function of the setting of the dentist's practice. CONCLUSION: The patient's race may influence a dentist's decision whether to extract or retain a decayed tooth. PMID- 16262616 TI - Avulsion of primary teeth and sequelae on the permanent successors. AB - The purpose of the present study was to determine the frequency of avulsion of primary teeth and the location of the avulsed tooth in a representative population of Danish children. Also, the frequency and the type of developmental disturbances in the permanent successors were assessed and related to age at the time of injury. The material included dental records of 4238 children from three clinics in Municipal Dental Health Services near Copenhagen, Denmark. The children were born between 1 January 1983 and 31 December 2000. Thirty-five children (0.8%) were identified as having avulsed in all 44 primary teeth most frequently the maxillary incisors (89%). Thirty-three fully erupted permanent successors were included in the study, the prevalence of developmental disturbances was 30% (10 teeth). The results showed the risk of developmental disturbances in the permanent successors to be more frequent the younger the age at the time of injury (P = 0.04). Discolouration affected all 10 permanent teeth, but also hypoplasia and horizontal enamel hypoplasia were found. PMID- 16262615 TI - Primary tooth injuries in Norwegian children (1-8 years). AB - This prospective study examined the yearly incidence of traumatic injuries to primary teeth. The aim of the study was to find out more about dental injuries to primary teeth in Norwegian children. The study was performed in one county of Norway involving approximately 20,000 children in the age group 1-8 years. Twenty seven public dental clinics and 42 dentists participated. The dentists attended information and calibration meetings and received illustrations of the classification of dental trauma as well as examples of how to fill in the forms correctly. Two hundred and sixty-six children were involved, including 447 primary teeth, recorded during a 1-year registration period (2003). The dental trauma incidence was 1.3% with 3.5-year old being the most accident-prone. Boys were significantly more often injured than girls, 164 versus 102 (P < 0.001). The upper central incisors were most involved (92%), with a non-significant difference between the right and left side. The minor periodontal injuries dominated (59%). Hard tissue injuries were far less frequent (13%). Avulsions were observed in 6.5% and intrusions in 7.5% of the injured children, being 5.5% and 5% of the injured teeth. Most of the injuries occurred either at home (38%) or at kindergarten (32%). Sixty-two percentage were falling accidents often sustained during children's play, and 25% were pure playing accidents. CONCLUSION: In Norwegian children aged 1-8 years, with a predominance of boys, the most common primary tooth injuries were minor luxations of the maxillary central incisors, sustained at an age of 3.5 years. As the traumas often occurred during children's play and/or were the consequence of falls, these are difficult to prevent. However, follow ups should be carried out to disclose pulpal or periodontal complications and/or developmental disturbances of the permanent successors. PMID- 16262617 TI - Physical education undergraduates and dental trauma knowledge. AB - The aim of the present study was to assess the level of knowledge of undergraduates from the College of Physical Education (Toledo, Aracatuba) concerning dental avulsion injuries. Data showed that 95% of the respondents did not know what dental avulsion is, 73.5% said they know how to define dental replantation, however, only 26% were able to do it correctly. When asked about first emergency measures after an avulsion, 50% of the respondents said they know what they should do, and the most cited measure was to seek a dentist. When asked about optimal storage media, 45.5% would keep it in a favorable one, and 28% did not know where to keep the tooth until treatment. Only 25.6% indicated a suitable extra-oral time for replantation; 90.3% of the respondents had received no advice about the emergency management of dental avulsion; 90% said they consider this an important and necessary subject. The results indicated that educational campaigns are necessary to improve the emergency management of dental injuries by those future P.E. professors for a better prognosis of dental replantation. PMID- 16262618 TI - Cytotoxicity analysis of alendronate on cultured endothelial cells and subcutaneous tissue. a pilot study. AB - The use of alendronate, a bisphosphonate which is able to inhibit bone resorption, in order to prevent dental root resorption after tooth replantation would be of clinical relevance. However, this drug must be biocompatible to the periapical tissues. The aim of this study was to analyze the effect of an alendronate paste in polyethyleneglycol (2 g ml(-1)) on endothelial cells in culture (in vitro) and on rat subcutaneous tissue (in vivo). For the in vitro study the paste was applied on round glass coverslips that were immersed into confluent cell cultures (clone Cips). The cell viability percentages of these cultures were obtained 0, 6 and 12 h after contact with the substance. As control, cultures that received plain coverslips were used. This analysis was carried out in triplicate using the Trypan blue dye exclusion assay. For the in vivo study the paste was introduced into polyethylene tubes that were placed into the rat subcutaneous tissue. The rats were killed 7 and 14 days later; then, the tissue sections stained with hematoxylin-eosin were analyzed. In vitro, the alendronate caused a significant decrease in the cell viability in 6 h (P < 0.05) and 12 h (P < 0.01), when compared with the control cultures. In vivo the tissue response was exuberant and similar at the two experimental times. There was a necrosis in a comprehensive area in contact with the open end of the tube. Presence of micro-abscesses and intense inflammatory infiltrate in the hypoderm permeating the muscle fibers and fat lobules were observed. In conclusion, the alendronate paste in polyethylene glycol as used showed to be highly cytotoxic in vitro as well as in vivo. PMID- 16262620 TI - Oral and dental complications of intra-oral piercing. AB - The present study assessed the prevalence of oral piercing among young adults and revealed the types and rate of complications following oral piecing, as well as the awareness of the complications. The study included 400 consecutive patients, who randomly arrived at a military dental office. Before dental examination, patients were requested to fill out a questionnaire regarding oral piercing, their awareness of its complications, and the occurrence of complications related to piercing. Intra-oral examination included special attention to piercing related complications, such as tooth fractures, gingivitis, bleeding, infections, gingival recessions, etc. A total of 389 patients, 210 (54%) males and 179 (46%) females agreed to participate (97.3% response rate), with an average age of 20.08 +/- 1.1 years. Of the participants, 79 (20.3%) reported having at least one type of oral piercing; lingual piercing was the most common. Swelling and bleeding after piercing were reported by 41 (51.9%) and 36 (45.7%) participants, respectively. Among the participants, 225 (57.8%) were unaware of the dangers of intra-oral piercing. Clinical examination revealed 15 fractured teeth in 11 (13.9%) participants with piercing. Gingival recessions were observed in 21 (26.6%), mostly in the mandibular incisor area. Dentists should be aware of the increasing number of patients with pierced intra- and peri-oral sites and to provide appropriate guidance to patients who contemplate body piercing involving oral sites. PMID- 16262619 TI - The use of enamel matrix derivative (Emdogain) for improvement of probing attachment level of the autotransplanted teeth. AB - The enamel matrix derivative (EMD, Emdogain) was used for the purpose to obtain the periodontal regeneration on the denuded root-surfaces of the donor teeth in two cases of the immediate tooth-transplantation. The root-surfaces at the cervical portion of the teeth were denuded because of extrusion. The healthy periodontium of each tooth remained at the apical portion of the roots. The denuded root-surfaces were cleansed before extraction. Then, the donor teeth were gently extracted with forceps, administered EMD, and transplanted so that the denuded surfaces were covered by gingival flaps. After the transplantation, the mean probing attachment level (PAL) improved 3.2 mm and 1.5 mm, respectively. The mean probing pocket depth was within normal level. The actual supporting areas of the roots of the transplanted teeth increased and the teeth worked as the abutments of prosthetic bridges. PMID- 16262621 TI - Is anti-resorptive regenerative therapy working in case of replantation of avulsed teeth? PMID- 16262623 TI - The SDF-1/CXCR4 pathway and the development of the cerebellar system. AB - Mice deficient for the chemokine receptor CXCR4 show premature translocation of granule cell neuroblasts from their germinal zone into the nascent cerebellum [Y. R. Zuo et al. (1998) Nature, 393, 595-599]. Here, we used CXCR4-null mice to analyse the early development of cerebellar cortical inhibitory interneurons and pontine neurons which, in the adult, are synaptically integrated with granule cells. Cortical inhibitory interneuronal precursors normally invade the cerebellar anlage of CXCR4-deficient mice, but their dispersal is impeded by dislocated foci of proliferating granule cells, from which they are excluded. This is reminiscent of the strict exclusion of inhibitory interneuronal precursors from the superficial external granule cell layer. As inhibitory interneuronal precursors readily mingle with post-mitotic granule cells both in wild-type and CXCR4-null mice, these findings indicate that the developmentally regulated interactions between granule and inhibitory interneuronal precursors are independent of SDF-1/CXCR4 signalling. In contrast, the transit of pontine neurons from the rhombic lip through the anterior extramural stream to the basilar pons is disrupted in CXCR4-deficient animals. Migrating pontine neurons express CXCR4, and in CXCR4-null animals these cells are found displaced deep into the brainstem. Consequently, nascent pontine nuclei in CXCR4-deficient animals are hypoplastic. Moreover, they fail to express plexin D1, suggesting that SDF-1/CXCR4 signalling may also impinge on axon guidance critical to the orderly formation of granule cell mossy fibre afferents. PMID- 16262624 TI - Eph/ephrin expression in the adult rat visual system following localized retinal lesions: localized and transneuronal up-regulation in the retina and superior colliculus. AB - Following unilateral optic nerve section in adult PVG hooded rat, the axon guidance cue ephrin-A2 is up-regulated in caudal but not rostral superior colliculus (SC) and the EphA5 receptor is down-regulated in axotomised retinal ganglion cells (RGCs). Changes occur bilaterally despite the retino-collicular projection being mostly crossed. Here we investigate the dynamics of Eph/ephrin expression using in situ hybridization and semi-quantitative immunohistochemistry after localized retinal lesions. Unilateral krypton laser lesions to dorso-nasal retina ablated contralaterally projecting RGCs (DN group); ventro-temporal lesions ablated contralaterally and ipsilaterally projecting RGCs (VT group). Lesions of the entire retina served as controls (Total group). Results are compared to normal animals in which tectal ephrin-A2 and retinal EphA5 are expressed, respectively, as shallow ascending rostro-caudal and naso-temporal gradients. In both SCs of DN and Total groups, tectal ephrin-A2 was up-regulated caudally; in the VT group, expression remained normal bilaterally. Unilateral collicular ablation indicated that bilateral changes in ephrin-A2 expression are mediated via intercollicular pathways. EphA5 expression in the VT group was elevated in the intact nasal region of experimental retinae. For each experimental group, EphA5 expression was also elevated in nasal retina of the opposite eye, resulting in uniform expression across the naso-temporal axis. Up regulation of ephrin-A2 in caudal, but not rostral, SC suggests the enhancement of developmental positional information as a result of injury. Bilateral increases in retinal EphA5 expression demonstrate that signals for up-regulation operate interocularly. The study demonstrates that signals regulating guidance cue expression are both localized and relayed transneuronally. PMID- 16262625 TI - The T1 domain of Kv1.3 mediates intracellular targeting to axons. AB - Shaker K+ channels play an important role in modulating electrical excitability of axons. Recent work has demonstrated that the T1 tetramerization domain of Kv1.2 is both necessary and sufficient for targeting of the channel to the axonal surface [Gu, C., Jan, Y.N. & Jan, L.Y. (2003) Science,301, 646-649]. Here we use a related channel, Kv1.3, as a model to investigate cellular mechanisms that mediate axonal targeting. We show that the T1 domain of Kv1.3 is necessary and sufficient to mediate targeting of the channel to the axonal surface in pyramidal neurons in slices of cortex from neonatal rat. The T1 domain is also sufficient to cause preferential axonal localization of intracellular protein, which indicates that the domain probably does not work through compartment-specific endocytosis or compartment-specific vesicle docking. To determine whether the T1 domain mediates axonal trafficking of transport vesicles, we compared the trafficking of vesicles containing green fluorescent protein-labelled transferrin receptor with those containing the same protein fused with the T1 domain in living cortical neurons. Vesicles containing the wild-type transferrin receptor did not traffic to the axon, in accord with previously published results; however, those containing the transferrin receptor fused to T1 did traffic to the axon. These results are consistent with the T1 domain of Kv1.3 mediating axonal targeting by causing transport vesicles to traffic to axons and they represent the first evidence that such a mechanism might underlie axonal targeting. PMID- 16262626 TI - Neurite guidance by the FnC repeat of human tenascin-C: neurite attraction vs. neurite retention. AB - The alternatively spliced fibronectin type-III repeat C of human tenascin-C (fnC) provides directional cues to elongating neurites in vitro. When given a choice at an interface with poly L-lysine (PLL), rat cerebellar granule neurites preferentially crossed onto fnC (defined herein as neurite attraction) whereas neurites originating on fnC preferentially remained on fnC (defined as neurite retention). Guidance motifs were further refined using synthetic peptides spanning the sequence of fnC. We found that a peptide with amino acid sequence DINPYGFTVSWMASE was sufficient to attract and retain neurites. Peptides with alterations in NPYG facilitated neurite retention but not attraction and, conversely, molecules with alterations in ASE facilitated neurite attraction but not retention. Hence neurite attraction and neurite retention mediated by fnC are separable events that can be independently regulated. This property may prove valuable for the strategic design of peptide reagents for use in strategies to facilitate directed axonal regrowth following CNS injury. PMID- 16262627 TI - Reduced extracellular space in the brain of tenascin-R- and HNK-1 sulphotransferase deficient mice. AB - Tenascin-R (TN-R), a large extracellular glycoprotein, is an important component of the adult brain's extracellular matrix (ECM); tenascin-C (TN-C) is expressed mainly during early development, while human natural killer 1 (HNK-1) is a sulphated carbohydrate epitope that attaches to these molecules, modifying their adhesive properties. To assess their influence on extracellular space (ECS) volume and geometry, we used the real-time iontophoretic method to measure ECS volume fraction alpha and tortuosity lambda, and diffusion-weighted magnetic resonance imaging (MRI) to measure the apparent diffusion coefficient of water (ADC(W)). Measurements were performed in vivo in the cortex and CA1 hippocampal region of TN-R-, TN-C- and HNK-1 sulphotransferase (ST)-deficient adult mice and their wild-type littermate controls. In both cortex and hippocampus, the lack of TN-R or HNK-1 sulphotransferase resulted in a significant decrease in alpha and lambda. Compared with controls, alpha in TN-R-/- and ST-/- mice decreased by 22 26% and 9-15%, respectively. MRI measurements revealed a decreased ADC(W) in the cortex, hippocampus and thalamus. ADC(W) reflected the changes in alpha; the decrease in lambda indicated fewer diffusion obstacles in the ECS, presumably due to a decreased macromolecular content. No significant changes were found in TN-C /- animals. We conclude that in TN-R-/- and ST-/- mice, which show morphological, electrophysiological and behavioural abnormalities, the ECS is reduced and its geometry altered. TN-R, as an important component of the ECM, appears to maintain an optimal distance between cells. The altered diffusion of neuroactive substances in the brain will inevitably affect extrasynaptic transmission, neuron glia interactions and synaptic efficacy. PMID- 16262628 TI - ATF3 expression precedes death of spinal motoneurons in amyotrophic lateral sclerosis-SOD1 transgenic mice and correlates with c-Jun phosphorylation, CHOP expression, somato-dendritic ubiquitination and Golgi fragmentation. AB - To obtain insight into the morphological and molecular correlates of motoneuron degeneration in amyotrophic lateral sclerosis (ALS) mice that express G93A mutant superoxide dismutase (SOD)1 (G93A mice), we have mapped and characterized 'sick' motoneurons labelled by the 'stress transcription factors' ATF3 and phospho-c Jun. Immunocytochemistry and in situ hybridization showed that a subset of motoneurons express ATF3 from a relatively early phase of disease before the onset of active caspase 3 expression and motoneuron loss. The highest number of ATF3-expressing motoneurons occurred at symptom onset. The onset of ATF3 expression correlated with the appearance of ubiquitinated neurites. Confocal double-labelling immunofluorescence showed that all ATF3-positive motoneurons were immunoreactive for phosphorylated c-Jun. Furthermore, the majority of ATF3 and phospho-c-Jun-positive motoneurons were also immunoreactive for CHOP (GADD153) and showed Golgi fragmentation. A subset of ATF3 and phosphorylated c Jun-immunoreactive motoneurons showed an abnormal appearance characterized by a number of distinctive features, including an eccentric flattened nucleus, perikaryal accumulation of ubiquitin immunoreactivity, juxta-nuclear accumulation of the Golgi apparatus and the endoplasmic reticulum, and intense Hsp70 immunoreactivity. These abnormal cells were not immunoreactive for active caspase 3. We conclude that motoneurons in ALS-SOD1 mice prior to their death and disappearance experience a prolonged sick phase, characterized by the gradual accumulation of ubiquitinated material first in the neurites and subsequently the cell body. PMID- 16262629 TI - Differential expression of cell fate determinants in neurons and glial cells of adult mouse spinal cord after compression injury. AB - Cellular responses after spinal cord injury include activation of astrocytes, degeneration of neurons and oligodendrocytes, and reactions of the ependymal layer and meningeal cells. Because it has been suggested that tissue repair partially recapitulates morphogenesis, we have investigated the expression of several developmentally prominent molecules after spinal cord injury of adult mice where neurogenesis does not occur after injury. Cell fate determinants Numb, Notch-1, Shh and BMPs are abundantly expressed during development but mostly decline in the adult. In the present study, we investigated whether these genes are triggered by spinal cord injury as a sign of attempted recapitulation of development. Expression of Numb, Notch, Shh, BMP2/4 and Msx1/2 was analysed in the adult mouse spinal cord after compression injury by in situ hybridization up to 1 month after injury. The mRNA expression levels of Notch-1, Numb, Shh, BMP4 and Msx2 increased in the grey matter and/or white matter and in the ependyma rostral and caudal to the lesion site after injury. However, BMP2 and Msx1 were not up-regulated. Combining immunohistochemistry of cell type-specific markers with in situ hybridization we found that all the up-regulated genes were expressed in neurons. Moreover, Numb, BMP4 and Msx2 were also expressed by GFAP positive astrocytes, while Shh was expressed by MBP-positive oligodendrocytes. In conclusion, the cell fate determinants Notch-1, Numb, Shh, BMP4 and Msx2 are expressed in neurons and/or glial cells after injury in a time-dependent manner, suggesting that these genes reflect to some extent an endogenous self-repair potential by recapitulating some features of development. PMID- 16262630 TI - Increased generation of granule cells in adult Bcl-2-overexpressing mice: a role for cell death during continued hippocampal neurogenesis. AB - Programmed cell death is an important mechanism during brain development in order to control neuronal cell numbers and to correctly form neuronal circuitries. Programmed cell death is also present in neurogenic regions of the adult brain, and a significant portion of the adult-born cells is eliminated during the first months of maturation. We here address the question whether overexpression of the anti-apoptotic protein Bcl-2 would improve the survival of neural progenitor cells and, as a consequence, increase neurogenesis in the adult hippocampus. Transgenic animals, which express human Bcl-2 under the neuron-specific enolase promoter (NSE-huBcl-2), show a significant reduction of apoptotic cells in the hippocampal granule cell layer to about half of the wild-type level. These apoptotic cells are almost exclusively found in the zone of hippocampal progenitor activity and frequently co-label with the neuronal progenitor marker doublecortin (DCX). The rate of adult neurogenesis is doubled in the dentate gyrus of Bcl-2-overexpressing mice as demonstrated by quantification of progenitor cells using DCX and new neurons using bromodeoxyuridine (BrdU)/neuronal nuclei antigen (NeuN) double-labelling. The effect of Bcl-2 is limited to the late phase of progenitor maturation, as proliferation and early phase progenitor cells were not affected. The increased level of neurogenesis leads to a significantly higher total number of granule cells in the dentate gyrus. These results underline the importance of developmental cell death during neurogenesis in the adult brain. PMID- 16262631 TI - Disruption of the neurogenic potential of the dentate gyrus in a mouse model of temporal lobe epilepsy with focal seizures. AB - Adult hippocampal neurogenesis is enhanced in response to multiple stimuli including seizures. However, the relationship between neurogenesis and the development of temporal lobe epilepsy (TLE) remains unclear. Unilateral intrahippocampal injection of kainate in adult mice models the morphological characteristics (e.g. neuronal loss, gliosis, granule cell dispersion and hypertrophy) and occurrence of chronic, spontaneous recurrent partial seizures observed in human TLE. We investigated the influence of a kainate-induced epileptogenic focus on hippocampal neurogenesis, comparing neural stem cell proliferation following status epilepticus and spontaneous recurrent partial seizures. Cell proliferation in the subgranular zone was transiently increased bilaterally after kainate treatment. As a result, neurogenesis was stimulated in the contralateral dentate gyrus. In contrast, the epileptic hippocampus exhibited a strongly reduced neurogenic potential, even after onset of spontaneous recurrent partial seizures, possibly due to an alteration of the neurogenic niche in the subgranular zone. These results show that neurogenesis does not contribute to the formation of the epileptic focus and may be affected when dispersion of dentate gyrus granule cells occurs. Therefore, in patients with TLE, hippocampal sclerosis and granule cell dispersion may play a significant role in disrupting the potential for hippocampal neurogenesis. PMID- 16262632 TI - The fate of neural progenitor cells expressing astrocytic and radial glial markers in the postnatal rat dentate gyrus. AB - In the dentate gyrus neurons continue to be generated from late embryonic to adult stage. Recent extensive studies have unveiled several key aspects of the adult neurogenesis, but only few attempts have so far been made on the analysis of the early postnatal neurogenenesis, a transition state between the embryonic and adult neurogenesis. Here, we focus on the early postnatal neurogenesis and examine the nature and development of neural progenitor cells in Wistar rats. Immunohistochemistry for Ki67, a cell cycle marker, and 5-bromo-2-deoxyuridine (BrdU) labelling show that cell proliferation occurs mainly in the hilus and partly in the subgranular zone. A majority of the proliferating cells express S100beta and astrocyte-specific glutamate transporter (GLAST) and the subpopulation are also positive for glial fibrillary acidic protein (GFAP) and nestin. Tracing with BrdU and our modified retrovirus vector carrying enhanced green fluorescent protein (GFP) indicate that a substantial population of the proliferating cells differentiate into proliferative neuroblasts and immature neurons in the hilus, which then migrate to the granule cell layer (66.8%), leaving a long axon-like process behind in the hilus, and the others mainly become star-shaped astrocytes (12.0%) and radial glia-like cells (4.7%) in the subgranular zone. These results suggest that the progenitors of the granule cells expressing astrocytic and radial glial markers, proliferate and differentiate into neurons mainly in the hilus during the early postnatal period. PMID- 16262633 TI - Contributions of protein phosphatases PP1, PP2A, PP2B and PP5 to the regulation of tau phosphorylation. AB - Abnormal hyperphosphorylation of tau is believed to lead to neurofibrillary degeneration in Alzheimer's disease (AD) and other tauopathies. Recent studies have shown that protein phosphatases (PPs) PP1, PP2A, PP2B and PP5 dephosphorylate tau in vitro, but the exact role of each of these phosphatases in the regulation of site-specific phosphorylation of tau in the human brain was unknown. Hence, we investigated the contributions of these PPs to the regulation of tau phosphorylation quantitatively. We found that these four phosphatases all dephosphorylated tau at Ser199, Ser202, Thr205, Thr212, Ser214, Ser235, Ser262, Ser396, Ser404 and Ser409, but with different efficiencies toward different sites. The K(m) values of tau dephosphorylation catalysed by PP1, PP2A and PP5 were 8-12 microm, similar to the intraneuronal tau concentration of human brain, whereas the K(m) of PP2B was fivefold higher. PP2A, PP1, PP5 and PP2B accounted for approximately 71%, approximately 11%, approximately 10% and approximately 7%, respectively, of the total tau phosphatase activity of human brain. The total phosphatase activity and the activities of PP2A and PP5 toward tau were significantly decreased, whereas that of PP2B was increased in AD brain. PP2A activity negatively correlated to the level of tau phosphorylation at the most phosphorylation sites in human brains. Our findings indicate that PP2A is the major tau phosphatase that regulates its phosphorylation at multiple sites in human brain. The abnormal hyperphosphorylation of tau is partially due to a downregulation of PP2A activity in AD brain. PMID- 16262634 TI - Ultrastructural evidence for a pre- and postsynaptic localization of full-length trkB receptors in substantia gelatinosa (lamina II) of rat and mouse spinal cord. AB - Brain-derived neurotrophic factor (BDNF) exerts its trophic effects by acting on the high-affinity specific receptor trkB. BDNF also modulates synaptic transmission in several areas of the CNS, including the spinal cord dorsal horn, where it acts as a pain modulator by yet incompletely understood mechanisms. Spinal neurons are the main source of trkB in lamina II (substantia gelatinosa). Expression of this receptor in dorsal root ganglion (DRG) cells has been a matter of debate, whereas a subpopulation of DRG neurons bears trkA receptors and contains BDNF. By the use of two different trkB antibodies we observed that 7.7% and 10.8% of DRG neurons co-expressed BDNF + trkB but not trkA, respectively, in rat and mouse. Ultrastructurally, full-length trkB (fl-trkB) receptors were present at somato-dendritic membranes of lamina II neurons (rat: 66.8%; mouse: 73.8%) and at axon terminals (rat: 33.2%; mouse: 26.2%). In both species, about 90% of these terminals were identified as primary afferent fibres (PAFs) considering their morphology and/or neuropeptide content. All fl-trkB immunopositive C boutons in type Ib glomeruli were immunoreactive for BDNF and, at individual glomeruli and axo-dendritic synapses, fl-trkB receptors were located in a mutually exclusive fashion at pre- or postsynaptic membranes. Thus, only a small fraction of fl-trkB-immunoreactive dendrites were postsynaptic to BDNF-immunopositive PAFs. This is the first ultrastructural description of fl trkB localization at synapses between first- and second-order sensory neurons in lamina II, and suggests that BDNF may be released by fl-trkB-immunopositive PAFs to modulate nociceptive input in this lamina of dorsal horn. PMID- 16262635 TI - Long-range oscillatory Ca2+ waves in rat spinal dorsal horn. AB - Synchronous activity of large populations of neurons shapes neuronal networks during development. However, re-emergence of such activity at later stages of development could severely disrupt the orderly processing of sensory information, e.g. in the spinal dorsal horn. We used Ca2+ imaging in spinal cord slices of neonatal and young rats to assess under which conditions synchronous activity occurs in dorsal horn. No spontaneous synchronous Ca2+ transients were detected. However, increasing neuronal excitability by application of 4-aminopyridine after pretreatment of the slice with blockers of (RS)-alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (AMPA)/kainate, gamma-aminobutyric acid (GABA)(A) and glycine receptors evoked repetitive Ca2+ waves in dorsal horn. These waves spread mediolaterally with a speed of 1.0 +/- 0.1 mm/s and affected virtually every dorsal horn neuron. The Ca2+ waves were associated with large depolarizing shifts of the membrane potential of participating neurons and were most likely synaptically mediated because they were abolished by blockade of action potentials or N-methyl-D-aspartate (NMDA) receptors. They were most pronounced in the superficial dorsal horn and absent from the ventral horn. A significant proportion of the Ca2+ waves spread to the contralateral dorsal horn. This seemed to be enabled by disinhibition as primary afferent-induced dorsal horn excitation crossed the midline only when GABA(A) and glycine receptors were blocked. Interestingly, the Ca2+ waves occurred under conditions where AMPA/kainate receptors were blocked. Thus, superficial dorsal horn NMDA receptors are able to sustain synchronous neuronal excitation in the absence of functional AMPA/kainate receptors. PMID- 16262636 TI - Involvement of spinal cord nuclear factor kappaB activation in rat models of proinflammatory cytokine-mediated pain facilitation. AB - Proinflammatory cytokines, such as interleukin-1beta and tumour necrosis factor alpha, are released by activated glial cells in the spinal cord and play a major role in pain facilitation. These cytokines exert their actions, at least partially, through the activation of the transcription factor, nuclear factor kappaB (NF-kappaB). In turn, NF-kappaB regulates the transcription of many inflammatory mediators, including cytokines. We have previously shown that intrathecal injection of the human immunodeficiency virus-1 (HIV-1) envelope glycoprotein, gp120, induces mechanical allodynia via the release of proinflammatory cytokines. Here, we investigated whether NF-kappaB is involved in gp120-induced pain behaviour in Sprague-Dawley rats. Intrathecal administration of NF-kappaB inhibitors, pyrrolidinedithiocarbamate (PDTC) and SN50, prior to gp120 partially attenuated gp120-induced allodynia. In addition, PDTC delayed and reversed allodynia in a model of neuropathic pain induced by sciatic nerve inflammation. These observations suggest that intrathecal gp120 may lead to activation of NF-kappaB within the spinal cord. To reveal NF-kappaB activation, we assessed inhibitory factor kappaBalpha (IkappaBalpha) mRNA expression by in situ hybridization, as NF-kappaB activation up-regulates IkappaBalpha gene expression as part of an autoregulatory feedback loop. No or low levels of IkappaBalpha mRNA were detected in the lumbar spinal cord of vehicle-injected rats, whereas IkappaBalpha mRNA expression was markedly induced in the spinal cord following intrathecal gp120 in predominantly astrocytes and endothelial cells. Moreover, IkappaBalpha mRNA expression positively correlated with proinflammatory cytokine protein levels in lumbosacral cerebrospinal fluid. Together, these results demonstrate that spinal cord NF-kappaB activation is involved, at least in part, in exaggerated pain states. PMID- 16262637 TI - Inputs to nucleus pontis caudalis from adjacent trigeminal areas. AB - Recent studies suggest that the nucleus pontis caudalis (nPontc) plays a role in patterning mastication through interactions with the adjacent lateral tegmentum. In this study, we used in vitro intracellular recording and staining to describe the basic membrane properties and morphology of nPontc neurones and to further explore interactions with adjacent structures, using coronal sections of the brainstem of 78 rats, aged 9-28 days. Neurones were large, with dendrites that spread in all directions, and about 64% fired tonically even in the absence of synaptic inputs. Tonic neurones were predominant in the centre of the nucleus. Electrical stimulation of all regions of the nPontc produced mixed excitatory and inhibitory effects on interneurones of lateral tegmental nuclei. Focal inactivation of the dorsal nPontc with injections of tetrodotoxin also had mixed effects on the spontaneous firing of both interneurones and motoneurones but similar injections in the ventral nPontc produced mostly increases of firing. Sixty-five percent of nPontc neurones received synaptic inputs from the lateral tegmental areas and most of these (68%) were excitatory and mediated by glutamatergic receptors. Inhibitory postsynaptic potentials were mediated by GABA(A) or glycinergic receptors. Although most responses occurred at relatively long latencies (> 2 ms), they could follow relatively high-frequency stimulation (> 50 Hz). Excitatory and inhibitory connections between ipsi- and contralateral nPontc neurones were also documented, which could contribute to bilateral coordination of jaw movements. This study provides evidence that the nPontc exerts both tonic and phasic influences on the premotor components of the masticatory central pattern generator. PMID- 16262638 TI - Histamine-immunoreactive neurons in the mouse and rat suprachiasmatic nucleus. AB - Among the well-established roles of the neurotransmitter histamine (HA) is that as a regulator of the sleep-wake cycle, which early gained HA a reputation as a 'waking substance'. The tuberomammillary nucleus (TMN) of the posterior hypothalamus, which contains the sole source of neuronal HA in the brain, is reciprocally connected to the suprachiasmatic nucleus (SCN) which, in turn, is best known as the pacemaker of circadian rhythms in mammals. We report HA immunoreactive (-ir) neurons in the mouse and rat SCN that neither display immunoreactivity (-iry) for the HA-synthesizing enzyme histidine decarboxylase (HDC) nor contain HDC mRNA. Further, HA-iry was absent in the SCN of HDC knockout mice, but present in appropriate control animals, indicating that the observed HA iry is HDC dependent. Experiments with hypothalamic slice cultures and i.c.v. injection of HA suggest that HA in the SCN neurons originates in the TMN and is transported from the TMN along histaminergic fibres known to innervate the SCN. These results could indicate the existence of a hitherto unknown uptake mechanism for HA into neurons. Through HA uptake and, putatively, re-release of the captured HA, these neurons could participate in the HA-mediated effects on the circadian system in concert with direct histaminergic inputs from the TMN to the SCN. The innervation of the SCN by several neurotransmitter systems could provide a way for other systems to affect the HA-containing neuronal cell bodies in the SCN. PMID- 16262640 TI - The spatial filtering properties of local edge detectors and brisk-sustained retinal ganglion cells. AB - We compared image computation in the rabbit retina by two different cell types: the so-called 'local edge detecting' ganglion cells and the well-known brisk sustained ganglion cells. From both anatomical and physiological evidence, these cells are present in nearly equal numbers and thus overlap to sample the same regions of visual space. We recorded simultaneously from overlapping cells on a dense microelectrode array. The results were analysed using an anatomically realistic simulation of the retina's processing levels. The 'local edge detecting' cell was found to be tuned to higher spatial frequencies and to have a narrower spatial frequency bandpass than the brisk-sustained cells. Simulation revealed that this is due primarily to the 'zero-crossing' detector implied by the definition of the local edge detector. The outputs of the simulations in response to complex images were analysed quantitatively. The results showed the population of local edge detectors to transmit a sparser code than the brisk sustained cells. PMID- 16262641 TI - Distinct properties of two major excitatory inputs to hippocampal pyramidal cells: a computational study. AB - The two main sources of excitatory input to CA1 pyramidal cells, the Schaffer collaterals (SC) and the perforant path (PP), target different regions of the dendritic tree. This spatial segregation may have important consequences for the way in which different inputs affect the activity of principal neurons. We constructed detailed biophysical models of CA1 pyramidal cells, incorporating a variety of active conductances, and investigated the ability of synapses located in different dendritic segments to elicit a somatic voltage response. Synaptic efficacy as seen by the soma was strongly dependent on the site of the synapse, with PP inputs being more severely attenuated than SC inputs. Variability within SC inputs, but not between SC inputs and PP inputs, could be eliminated by appropriate scaling of synaptic efficacy. The spatial and temporal summation of multiple synaptic inputs was also investigated. While summation of SC inputs was linear up to the somatic spike threshold, PP inputs summed in a strongly sublinear fashion, with the somatic response remaining subthreshold even following the simultaneous activation of a large number of synapses and during stimulation with high-frequency trains. Finally, the relative impact of different pathways on somatic activity could be effectively altered by modulating the kinetic properties of dendritic transient K+ channels, corresponding to the activation of ascending modulatory neurotransmitter systems. In this case, the efficacy of the PP was enhanced by the dendritic generation and limited spread of action potentials. Strong PP activation could also evoke dendritic Ca++ spikes, which often triggered a somatic burst. PMID- 16262639 TI - Remodelling of spinal nociceptive mechanisms in an animal model of monoarthritis. AB - Intra-articularly injected complete Freund's adjuvant creates in rats a chronic monoarthritis suitable for studying neuronal plasticity and chronic pain. Using such a model, we report electrophysiological and morphological evidence of alterations in somatosensory synaptic function. In arthritic rats, the baseline activity of dorsal spinal cord wide dynamic range or nociceptive-specific neurons was greater than in control animals. Moreover, neuronal responses elicited by an innocuous stimulation with von Frey filaments applied to the arthritic joint were greater in amplitude and produced the afterdischarge that normally characterizes a nociceptive response. In contrast to the response in control animals, passive movement of the arthritic joint produced an increase in the amplitude of the response of these neurons to iontophoretic application of glutamate receptor agonists over a time frame of 10-30 min. This potentiation was blocked by pretreatment with a neurokinin-1 (NK-1) receptor antagonist, suggesting the involvement of substance P. Ultrastructural analysis of the dorsal horn revealed that movement of the arthritic joint also induced NK-1 receptor internalization, indicative of nociception. Morphological examination revealed significantly increased expression of substance P and its receptor within the superficial dorsal horn of monoarthritic animals. These unique functional and chemical changes reflect alterations in both presynaptic and postsynaptic mechanisms in nociceptive transmission at the spinal level. Thus, although treatment of arthritis should obviously target its peripheral aetiology, targeting its central components is a logical therapeutic complementary objective. PMID- 16262642 TI - Dynamics of directional selectivity in MT receptive field centre and surround. AB - We studied receptive field organization of motion-sensitive neurons in macaque middle temporal cortical area (MT), by mapping direction selectivity in space and in time. Stimuli consisted of pseudorandom sequences of single motion steps presented simultaneously at many different receptive field locations. Spatio temporal receptive field profiles were constructed by cross-correlating stimuli and spikes. The resulting spike-triggered averages revealed centre-surround organization. The temporal dynamics of the receptive fields were generally biphasic with increased probability for the preferred direction at short latency (50-70 ms) and decreased probability at longer latency (80-100 ms). The response latency of the receptive field surround was on average 16 ms longer than that of the centre. Our results show that surround input and biphasic behaviour reflect two different mechanisms, which make MT cells specifically sensitive to motion contrast in space and time. PMID- 16262643 TI - Striatal dopaminergic responses observed in latent inhibition are dependent on the hippocampal ventral subicular region. AB - We showed recently that behavioural and striatal dopaminergic (DA) responses obtained in latent inhibition are crucially dependent on the parahippocampal region, the entorhinal cortex. In the present study, we investigated the influence exerted by the hippocampal ventral subicular region (SUB) on the DA responses in the anterior part of the dorsal striatum using in vivo voltammetry in freely moving rats and the same latent inhibition paradigm. To that end, the left SUB was temporarily blocked with tetrodotoxin (TTX) during pre-exposure to a new olfactory stimulus (banana odour). During the second session the animals were aversively conditioned to banana odour. With respect to the results obtained during the test session (third presentation of banana odour), similar changes in behaviour and DA levels were obtained in control and conditioned rats microinjected with the solvent, phosphate-buffered saline (PBS), in the SUB, consistently with a latent inhibition phenomenon. In contrast, after reversible inactivation of the SUB during the pre-exposure session, TTX-pre-exposed conditioned animals displayed aversive behaviour in the test session, and anterior striatal DA variations in these animals differed significantly from those obtained in pre-exposed rats injected locally with PBS. Striatal DA variations obtained in conditioned animals microinjected with TTX were also significantly different from those observed in conditioned non-pre-exposed animals. The present data suggest that, in parallel to the entorhinal cortex, the SUB regulates the latent inhibition-related behavioural and DA responses in the anterior part of the dorsal striatum. These data may provide new insight into the pathophysiology of schizophrenic psychoses. PMID- 16262644 TI - Time-sensitive enhancement of motor learning with the less-affected forelimb after unilateral sensorimotor cortex lesions in rats. AB - Unilateral damage to the forelimb region of the sensorimotor cortex (FLsmc) results in time-dependent changes in neuronal activity, structure and connectivity in the contralateral motor cortex of adult rats. These changes have been linked to facilitation of motor skill learning in the less affected/ipsilesional forelimb, which is likely to promote its use in the development of behavioral compensation. The goal of this study was to determine whether an early post-lesion-sensitive time period exists for this enhanced learning and whether it is linked to synaptogenesis in the contralesional motor cortex. Rats were trained for 21 days on a skilled reaching task with the ipsilesional forelimb beginning 4 or 25 days after unilateral ischemic (endothelin-1-induced) FLsmc lesions or sham operations. As found previously, reaching performance was significantly enhanced in rats trained early post-lesion compared with sham-operates. In rats trained later post-lesion, performance was neither significantly different from time-matched sham-operates nor strikingly different from animals trained earlier post-lesion. In layer V of the contralesional motor cortex, stereological methods for light and electron microscopy revealed significantly more total, multisynaptic bouton and perforated synapses per neuron compared with sham-operates, but there were no significant differences between early- and late-trained lesion groups. Thus, there appears to be a sensitive time window for the maximal expression of the enhanced learning capacity of the less-affected forelimb but this window is broadly, rather than sharply, defined. These results indicate that relatively long-lasting lesion induced neuronal changes are likely to underlie the facilitation of learning with the less-affected forelimb. PMID- 16262645 TI - Cognitive disorders and neurogenesis deficits in Huntington's disease mice are rescued by fluoxetine. AB - Huntington's disease (HD) is a neurodegenerative disorder caused by an expanded CAG trinucleotide repeat encoding an extended polyglutamine tract in the huntingtin protein. Affected individuals display progressive motor, cognitive and psychiatric symptoms (including depression), leading to terminal decline. Given that transgenic HD mice have decreased hippocampal cell proliferation and that a deficit in neurogenesis has been postulated as an underlying cause of depression, we hypothesized that decreased hippocampal neurogenesis contributes to depressive symptoms and cognitive decline in HD. Fluoxetine, a serotonin-reuptake inhibitor commonly prescribed for the treatment of depression, is known to increase neurogenesis in the dentate gyrus of wild-type mouse hippocampus. Here we show that hippocampal-dependent cognitive and depressive-like behavioural symptoms occur in HD mice, and that the administration of fluoxetine produces a marked improvement in these deficits. Furthermore, fluoxetine was found to rescue deficits of neurogenesis and volume loss in the dentate gyrus of HD mice. PMID- 16262646 TI - Putaminal gray matter volume decrease in panic disorder: an optimized voxel-based morphometry study. AB - Our study aimed to identify gray matter volume differences between panic disorder patients and healthy volunteers using optimized voxel-based morphometry. Gray matter volume was compared between 18 panic subjects and 18 healthy volunteers. Panic disorder severity scale (PDSS) and Zung self-rating anxiety scale (Z-SAS) were administered. Gray matter volumes of bilateral putamen were decreased in panic subjects relative to healthy comparison subjects (corrected P < 0.05). Decreased gray matter volume was also observed in the right precuneus, right inferior temporal gyrus, right inferior frontal gyrus, left superior temporal gyrus, and left superior frontal gyrus at a less conservative level of significance. PDSS score negatively correlated with gray matter volume in the left putamen, right putamen, right inferior frontal gyrus, and left superior frontal gyrus in panic subjects. The duration of illness negatively correlated with left putaminal gray matter volume. There was also a negative correlation between gray matter volume in right putamen and Z-SAS score in panic subjects. The current study reports a putaminal gray matter volume decrease in panic subjects, which may be related to the clinical severity of panic disorder. PMID- 16262647 TI - Distinct and overlapping fMRI activation networks for processing of novel identities and locations of objects. AB - The ventral visual stream processes information about the identity of objects ('what'), whereas the dorsal stream processes the spatial locations of objects ('where'). There is a corresponding, although disputed, distinction for the ventrolateral and dorsolateral prefrontal areas. Furthermore, there seems to be a distinction between the anterior and posterior medial temporal lobe (MTL) structures in the processing of novel items and new spatial arrangements, respectively. Functional differentiation of the intermediary mid-line cortical and temporal neocortical structures that communicate with the occipitotemporal, occipitoparietal, prefrontal, and MTL structures, however, is unclear. Therefore, in the present functional magnetic resonance imaging (fMRI) study, we examined whether the distinction among the MTL structures extends to these closely connected cortical areas. The most striking difference in the fMRI responses during visual presentation of changes in either items or their locations was the bilateral activation of the temporal lobe and ventrolateral prefrontal cortical areas for novel object identification in contrast to wide parietal and dorsolateral prefrontal activation for the novel locations of objects. An anterior-posterior distinction of fMRI responses similar to the MTL was observed in the cingulate/retrosplenial, and superior and middle temporal cortices. In addition to the distinct areas of activation, certain frontal, parietal, and temporo-occipital areas responded to both object and spatial novelty, suggesting a common attentional network for both types of changes in the visual environment. These findings offer new insights to the functional roles and intrinsic specialization of the cingulate/retrosplenial, and lateral temporal cortical areas in visuospatial cognition. PMID- 16262648 TI - Kappa- and delta-opioid receptor functional activities are increased in the caudate putamen of cannabinoid CB1 receptor knockout mice. AB - The purpose of this study was to examine the functional interaction between endogenous opioid and cannabinoid receptor systems in the caudate putamen and nucleus accumbens. We therefore examined by autoradiography the functional activity and density of micro-, kappa- and delta-opioid receptors in both brain regions of cannabinoid CB1 receptor knockout mice. Functional activity was estimated by measuring agonist-stimulated [35S]GTPgammaS binding. Results showed that deletion of the CB1 cannabinoid receptor markedly increased kappa-opioid (50%) and delta-opioid (42%) receptor activities whereas no differences were found in micro-opioid receptor in the caudate putamen. In contrast, binding autoradiography showed a similar density of micro-, kappa- and delta-opioid receptors between mutant and wild-type mice. No differences were found in densities or activities of micro-, kappa- and delta-opioid receptors between mutant and wild-type mice in the nucleus accumbens. Taken together, our results revealed that deletion of CB1 cannabinoid receptors produced a pronounced increase in the activity of kappa- and delta-opioid receptors in the caudate putamen. This endogenous interaction between opioid and cannabinoid receptors may be relevant to further understand a variety of neuroadaptative processes involving the participation of opioid receptors, such as motor behaviour, emotional responses and drug dependence. PMID- 16262649 TI - Sleep deprivation suppresses neurogenesis in the adult hippocampus of rats. AB - We reported previously that 96 h of sleep deprivation (SD) reduced cell proliferation in the dentate gyrus (DG) of the hippocampus in adult rats. We now report that SD reduces the number of new cells expressing a mature neuronal marker, neuronal nuclear antigen (NeuN). Rats were sleep-deprived for 96 h, using an intermittent treadmill system. Total sleep time was reduced to 6.9% by this method in SD animals, but total treadmill movement was equated in SD and treadmill control (CT) groups. Rats were allowed to survive for 3 weeks after 5 bromo-2-deoxyuridine (BrdU) injection. The phenotype of BrdU-positive cells in the DG was assessed by immunofluorescence and confocal microscopy. After 3 weeks the number of BrdU-positive cells was reduced by 39.6% in the SD group compared with the CT. The percentage of cells that co-localized BrdU and NeuN was also lower in the SD group (SD: 46.6 +/- 1.8% vs. CT: 71.9 +/- 2.1, P < 0.001). The percentages of BrdU-labeled cells co-expressing markers of immature neuronal (DCX) or glial (S100-beta) cells were not different in SD and CT groups. Thus, SD reduces neurogenesis in the DG by affecting both total proliferation and the percentage of cells expressing a mature neuronal phenotype. We hypothesize that sleep provides anabolic or signaling support for proliferation and cell fate determination. PMID- 16262650 TI - Relaxin receptor activation in the basolateral amygdala impairs memory consolidation. AB - The peptide-hormone relaxin has well-established actions in male and female reproductive tracts, and has functional effects in circumventricular regions of brain involved in neurohormonal secretion. In the current study, we initially mapped the distribution of mRNA encoding the relaxin receptor--leucine-rich repeat-containing G-protein-coupled receptor 7 (LGR7)- and [33P]-human relaxin binding sites in extra-hypothalamic sites of male Sprague-Dawley rats. The basolateral amygdala (BLA) expressed high levels of LGR7 mRNA and relaxin-binding sites and, although relaxin peptide was not detected in the BLA, several brain regions that send projections to the BLA were found to contain relaxin-expressing neurons. As it is well established that the BLA is involved in regulating the consolidation of memory for emotionally arousing experiences, we investigated whether activation of LGR7 in the BLA modulated memory consolidation for aversively motivated inhibitory avoidance training. Bilateral infusions of human relaxin (10-200 ng in 0.2 microL) into the BLA immediately after inhibitory avoidance training impaired 48-h retention performance in a dose-dependent manner. Delayed infusions of relaxin into the BLA 3 h after training were ineffective, indicating that the retention impairment was due to influences on memory consolidation. Post-training infusions of relaxin into the adjacent central amygdala, which is devoid of LGR7, did not impair retention. These findings suggest a novel function for endogenous relaxin-LGR7 signalling in rat brain involving regulation of memory consolidation. PMID- 16262651 TI - Clinical application of neurotrophic factors: the potential for primary auditory neuron protection. AB - Sensorineural hearing loss, as a result of damage to or destruction of the sensory epithelia within the cochlea, is a common cause of deafness. The subsequent degeneration of the neural elements within the inner ear may impinge upon the efficacy of the cochlear implant. Experimental studies have demonstrated that neurotrophic factors can prevent this degeneration in animal models of deafness, and can even provide functional benefits. Neurotrophic factor therapy may therefore provide similar protective effects in humans, resulting in improved speech perception outcomes among cochlear implant patients. There are, however, numerous issues pertaining to delivery techniques and treatment regimes that need to be addressed prior to any clinical application. This review considers these issues in view of the potential therapeutic application of neurotrophic factors within the auditory system. PMID- 16262652 TI - Expression of netrin-1, slit-1 and slit-3 but not of slit-2 after cerebellar and spinal cord lesions. AB - To determine whether members of the Netrin-1 and Slit families and their receptors are expressed after central nervous system (CNS) injury, we performed in situ hybridization for netrin-1, slit-1, 2 and 3, and their receptors (dcc, unc5h-1, 2 and 3, robo-1, 2 and 3) 8 days, 2-3 months and 12-18 months after traumatic lesions of rat cerebellum. The expression pattern of these molecules was unchanged in axotomized Purkinje cells, whereas unc5h3 expression was upregulated in deafferented granule cells. Cells expressing slit-2 or dcc were never detected at the lesion site. By contrast, cells expressing netrin-1, slit-1 and slit-3, unc5h-1, 2 and 3, and robo-1, 2 and 3 (rig-1) could be detected at the cerebellar lesion site as soon as 8 days after injury. Expression of unc5h-2, robo-1, robo-2, slit-1 and slit-3 at the lesion site was maintained until 3 months, and up to 12-18 months for unc5h-1 and 3 and robo-3. Likewise, in the mouse spinal cord, netrin-1, slit-1 and slit-3 were also expressed at the lesion site 8 days after injury. Most of the cells expressing these mRNAs were located at the centre of the lesions, suggesting that they are macrophages/activated microglial cells (macrophagic cells) or meningeal fibroblastic cells. The macrophagic nature of most Netrin-1-positive cells and the macrophagic or fibroblastic nature of Robo-1-positive cells were corroborated by double staining. Thus, Netrin-1, Slits and their receptors may contribute to the regenerative failure of axons in the adult CNS by inhibiting axon outgrowth or by participating in the formation of the CNS scar. PMID- 16262653 TI - Neuronal expression of Nogo-A mRNA and protein during neurite outgrowth in the developing rat olfactory system. AB - The major impediments to axonal regeneration in the central nervous system are growth-inhibitory proteins present in the myelin sheath, and Nogo-A is one of the most potent inhibitors synthesized by oligodendrocytes. However, neuronal expression of Nogo-A during development suggests that it may have an additional role. The spatio-temporal regulation of both Nogo-A mRNA and protein expression was examined by in situ hybridization and immunohistochemistry in the developing rat olfactory system. During embryonic and postnatal development (from E13 to P6), Nogo-A mRNA and protein were strongly expressed by differentiating neurons in the olfactory epithelium and in the olfactory bulb. From the second postnatal week, a progressive down-regulation of both Nogo-A mRNA and protein occurred, such that only a weak expression persisted in the adult olfactory system. Using double-immunostainings in the adult olfactory epithelium, we determined that Nogo A was preferentially expressed by immature olfactory receptor neurons extending axonal processes toward the olfactory bulb. At all developmental stages, Nogo-A protein was preferentially targeted in olfactory axons emerging from the olfactory epithelium. Using an in vitro model of olfactory axon growth, we demonstrated that, in addition to its presence along the entire axon length, Nogo A accumulated in axonal growth cone and at axonal branching points, with a distribution similar to that of microtubule-associated proteins. Moreover, Nogo-A was transiently expressed in dendritic processes in the postnatal olfactory bulb. Together, our data suggest that, in non-pathological conditions, Nogo-A may be involved in the processes of axonal growth and dendritic modeling through the regulation of microtubule dynamics. PMID- 16262654 TI - Identification of an ectodomain within the LAR protein tyrosine phosphatase receptor that binds homophilically and activates signalling pathways promoting neurite outgrowth. AB - Elucidation of mechanisms by which receptor protein tyrosine phosphatases (PTPs) regulate neurite outgrowth will require characterization of ligand-receptor interactions and identification of ligand-induced signalling components mediating neurite outgrowth. The first identified ligand of the leucocyte common antigen related (LAR) receptor PTP consists of a 99-residue ectodomain isoform, termed LARFN5C, which undergoes homophilic binding to LAR and promotes neurite outgrowth. We employed peptide mapping of LARFN5C to identify an active neurite promoting domain of LAR. A peptide mimetic consisting of 37 residues (L59) and corresponding to the fifth LAR fibronectin type III (FNIII) domain prevented LARFN5C homophilic binding, demonstrated homophilic binding to itself and promoted neurite outgrowth of mouse E16-17 hippocampal neurons and of dorsal root ganglia explants. Response to L59 was partially lost when using neurons derived from LAR-deficient (-/-) mice or neurons treated with LAR siRNA, consistent with homophilic interaction of L59 with LAR. L59 neurite-promoting activity was decreased in the presence of inhibitors of Src, Trk, PLCgamma, PKC, PI3K and MAPK. L59 activated Src (a known substrate of LAR), FAK and TrkB and also activated downstream signalling intermediates including PKC, ERK, AKT and CREB. BDNF augmented the maximal neurite-promoting activity of L59, a finding consistent with the presence of shared and distinct signalling pathways activated by L59 with BDNF and L59 with TrkB. These studies are the first to identify an ectodomain of LAR (located within the fifth FNIII domain) capable of promoting neurite outgrowth and point to novel approaches for promotion of neurite outgrowth. PMID- 16262655 TI - Opposite impacts of tenascin-C and tenascin-R deficiency in mice on the functional outcome of facial nerve repair. AB - The glycoproteins tenascin-C (TNC) and tenascin-R (TNR) are extracellular matrix proteins involved in the development, plasticity and repair of the nervous system. Altered expression patterns after nerve lesions in adult animals have suggested that these molecules influence axonal regeneration. To test this hypothesis, we investigated adult mice constitutively deficient in the expression of TNC, TNR or both, using the facial nerve injury paradigm. Quantitative analysis of vibrissal movements prior to nerve transection and repair (facial facial anastomosis) did not reveal genotype-specific differences, and thus impacts of the mutations on motor function in intact animals. Two months after nerve repair, recovery of vibrissal whisking was poor in wild-type mice, a typical finding after facial-facial anastomosis in rodents. Differential effects of the mutations on whisking were found: recovery of function was worse in TNC deficient and better in TNR null mice compared with wild-type littermates. In double-knockout animals, vibrissal performance was insufficient, but to a lesser extent compared with TNC null mutant mice. Retrograde labelling of motoneurons in the same animals showed that similar numbers of motoneurons had reinnervated the whisker pads in all experimental groups precluding varying extents of motoneuron death and/or axon regeneration failures as causes for the different outcomes of nerve repair. Our results provide strong evidence that TNC promotes and TNR impedes recovery after nerve lesion. These findings are of particular interest with regard to the scanty knowledge about factors determining success of regeneration in the peripheral nervous system of mammals. PMID- 16262656 TI - Photic inhibition of TrkB/Ras activity in the pigeon's tectum during development: impact on brain asymmetry formation. AB - Asymmetric photic stimulation during embryonic or post-hatch development induces a functional lateralization of the pigeon's visual system, which is accompanied by left-right differences in tectal cell sizes. The intracellular membrane anchored GTPase Ras can be activated by a number of upstream mechanisms including binding of brain-derived neurotrophic factor to its specific TrkB receptor. Ras activity plays an important morphogenetic role in neurons and therefore might also be involved in the asymmetric differentiation of tectal cells. To investigate the role of Ras, we determined the relative levels of activated Ras and of signalling active phospho-TrkB in tecta of light- and dark-incubated pigeons and combined this with an immunohistochemical detection of Ras-GTP and TrkB receptors. While Ras activation levels did not differ between light- and dark-incubated pigeons during embryonic development, directly after hatching Ras activity was significantly decreased in the stronger stimulated left tectum of light-incubated animals. This was accompanied by lower levels of TrkB phosphorylation. Immunohistochemical staining revealed Ras-GTP-positive cell bodies within the efferent cell layer. These cells were TrkB-positive and developed enlarged soma sizes within the right tectum during the first week after hatching. This association suggests asymmetric Ras activation to be involved in the asymmetric differentiation of the efferent cells as a result of asymmetric TrkB signalling. Because asymmetric light exposure occurs only during embryonic development, the observed transient asymmetric inhibition of TrkB/Ras activity after hatching may reflect differential embryonic maturation of tectal inhibitory circuits leading to a functional superiority of the right eye in the adult organism. PMID- 16262658 TI - Stimulation of PGE receptors EP2 and EP4 protects cultured neurons against oxidative stress and cell death following beta-amyloid exposure. AB - Alzheimer's disease (AD) is associated with gliosis, neuroinflammation and higher levels of prostaglandins. Conflicting roles for cyclooxygenases and prostaglandins in the etiopathology of AD have been reported. We hypothesized that PGE2 signaling through EP2 and EP4 G-protein-coupled receptors could protect against amyloid beta-peptide (Abeta) neurotoxicity by increasing the cAMP signaling cascade. Using primary neuronal cultures, we investigated the presence of EP receptors (EP1-4) and the action of PGE2 and EP receptor agonists on neuronal susceptibility to Abeta1-42 toxicity. Low concentrations (1 microm) of PGE2, butaprost (EP2 agonist), and 1-hydroxy-PGE1 (EP4/EP3 agonist) were neuroprotective against Abeta1-42 toxicity, while sulprostone (EP3/EP1 agonist) at similar doses had no detectable effects. EP2 and EP4 receptor-mediated neuroprotection would involve changes in cAMP levels, as both EP2 and EP4 agonists increased intracellular cAMP concentration by approximately doubling basal levels, and both exhibited neuroprotective actions against Abeta-induced toxicity. The protein kinase A (PKA) inhibitor RpcAMPS significantly attenuated the neuroprotection by butaprost, but not that by 1-hydroxy-PGE1, implying differences between EP2 and EP4 receptor protective mechanisms. Additionally, the increase in reactive oxygen species generated following exposure to Abeta was reduced by stimulation of both EP2 and EP4 receptors. Together, these results indicate that PGE2 can protect neurons against Abeta toxicity by acting on given receptors and stimulating a cascade of intracellular events, including the cAMP PKA pathway. We propose that development and testing of specific PGE2 receptor agonists downstream of cyclooxygenase could lead to therapeutic applications. PMID- 16262657 TI - Dynamic association of the Ca2+ channel alpha1A subunit and SNAP-25 in round or neurite-emitting chromaffin cells. AB - Although the specific interaction between synaptic protein SNAP-25 and the alpha1A subunit of the Cav2.1 channels, which conduct P/Q-type Ca2+ currents, has been confirmed in in vitro-translated proteins and brain membrane studies, the question of how native proteins can establish this association in situ in developing neurons remains to be elucidated. Here we report data regarding this interaction in bovine chromaffin cells natively expressing both proteins. The two carboxyl-terminal splice variants of the alpha1A subunit identified in these cells share a synaptic protein interaction ('synprint') site within the II/III loop segment and are immunodetected by a specific antibody against bovine alpha1A protein. Moreover, both alpha1A isoforms form part of the P/Q-channels-SNARE complexes in situ because they are coimmunoprecipitated from solubilized chromaffin cell membranes by a monoclonal SNAP-25 antibody. The distribution of alpha1A and SNAP-25 was studied in round or transdifferentiated chromaffin cells using confocal microscopy and specific antibodies: the two proteins are colocalized at the cell body membrane in both natural cell types. However, during the first stages of the cell transdifferentiation process, SNAP-25 migrates alone out to the developing growth cone and what will become the nerve endings and varicosities of the mature neurites; alpha1A follows and colocalizes to SNAP-25 in the now mature processes. These observations lead us to propose that the association between SNAP-25 and alpha1A during neuritogenesis might promote not only the efficient coupling of the exocytotic machinery but also the correct insertion of P/Q-type channels at specialized active zones in presynaptic neuronal terminals. PMID- 16262659 TI - Rapid modulatory effect of estradiol on acetylcholine-induced Ca2+ signal is mediated through cyclic-GMP cascade in LHRH-releasing GT1-7 cells. AB - Hypothalamic luteinizing hormone-releasing hormone neurons (LHRH) form the final pathway for the central control of reproduction through the release of LHRH into the pituitary-hypothalamic system. We previously found that LHRH-producing GT1-7 cells respond to acetylcholine (ACh) with an increase in intracellular calcium ([Ca2+]i) through activation of muscarinic receptors. This effect is acutely modulated by 17beta-estradiol in a manner compatible with specific membrane binding sites. Because increasing evidence suggests that second messengers are involved in the rapid action of estradiol, the aim of the present study was to identify the pathway underlying estrogen actions on ACh-induced Ca2+ signals. 8 Bromoguanosine 3',5'-cyclic monophosphate (10 microm) and C-type natriuretic peptide (10 microm) mimicked the effect of estradiol. On the contrary, neither dibutyryl cAMP (100 microm), forskolin (100 nm or 10 microm), or sodium nitroprusside (10 microm) induced any modification of [Ca2+]i in response to ACh. The effect of estradiol on calcium transients was totally blocked by two different cGMP-dependent protein kinase (PKG) inhibitors. In addition, phosphorylation of inositol 1,4,5-triphosphate (IP3) receptor was rapidly induced by estradiol but totally blocked when the cells were pretreated with a PKG inhibitor. We conclude that physiological concentrations of estradiol reduce ACh induced Ca2+ transients via a mechanism involving a membrane-associated guanylate cyclase, which finally induces a PKG-dependent IP3 receptor phosphorylation that modifies calcium release from the endoplasmic reticulum. PMID- 16262660 TI - Cell type-specific regulation of RE-1 silencing transcription factor (REST) target genes. AB - RE-1 silencing transcription factor (REST) is a transcriptional repressor that represses neuronal gene transcription in non-neuronal cells. REST target genes are expressed in neurons and in neuroendocrine cells. Here, we show that treatment with the histone deacetylase inhibitor trichostatin A (TSA) or expression of a mutant of REST (DP-REST:ER) that contains a transcriptional activation domain enhanced expression of the REST target genes encoding synaptophysin and secretogranin II in neuronal as well as in neuroendocrine cells. These data indicate that the synaptophysin and secretogranin II genes are similarly regulated in neuronal and neuroendocrine cells. In contrast, expression of the connexin36 gene was inducible by TSA or DP-REST:ER only in pancreatic alpha and beta cells, but not in neuronal and pituitary cells, indicating that transcriptional repression by REST functions in a cell type-specific manner. Expression of the BDNF and GluR2 genes, both described as targets of REST, was not induced by either TSA or expression of DP-REST:ER in neuronal or neuroendocrine cells. Chromatin immunoprecipitation experiments using antibodies directed against methylated histone H3Lys4 or H3Lys9 showed a perfect correlation between expression of REST target genes in different cell types and nucleosomal modifications that distinguish active from inactive genes. We conclude that the cell type-specific microenvironment, in particular the cell type-specific structure of the chromatin, is crucial for the ability of REST to control gene transcription. PMID- 16262663 TI - Vagal pulmonary afferents and central respiratory effects of 5-HT in newborn rats. AB - In decerebrate newborn rats, serotonin (5-HT) is a respiratory depressant via activation of 5-HT2 receptors, whereas it evokes respiratory stimulant effects when applied to the isolated brainstem obtained from the newborn rat. This discrepancy could be due to deafferentation in the in vitro preparation. The aim of our study was to analyse the role of vagal afferents in the modulation of central respiratory effects of 5-HT. In decerebrate cervically or abdominally bivagotomized newborn rats aged between 0 and 3 days, we recorded electrical activity from the diaphragm and from a hypoglossally innervated tongue muscle, as well as cardiac frequency (Fc), before and after application of 5-HT to the floor of the IVth ventricle. The effects of related agents (a 5-HT1A agonist, 8-OH DPAT, and a 5-HT2 agonist, DOI) were studied in cervically bivagotomized animals. For comparison, and to assess the spontaneous variability in inspiratory frequency (Fi) and Fc, sham groups were studied. Each group comprised ten newborn rats. In cervically bivagotomized newborn rats, 5-HT induces a significant increase in Fi, which is the opposite to that observed in decerebrate newborn rats with intact vagi. This respiratory effect is mediated in particular, via activation of 5-HT1A. By contrast, in abdominally bivagotomized newborn rats, a decrease in Fi was observed in response to 5-HT (as previously described in decerebrate animals with intact vagi). We conclude that pulmonary vagal afferents modulate the central respiratory action of 5-HT in decerebrate newborn rats, explaining the conflicting results between in vivo and in vitro experiments. PMID- 16262662 TI - Central nervous system neurons acquire mast cell products via transgranulation. AB - Resting and actively degranulating mast cells are found on the brain side of the blood-brain barrier. In the periphery, exocytosis of mast cell granules results in the release of soluble mediators and insoluble granule remnants. These mast cell constituents are found in a variety of nearby cell types, acquired by fusion of granule and cellular membranes or by cellular capture of mast cell granule remnants. These phenomena have not been studied in the brain. In the current work, light and electron microscopic studies of the medial habenula of the dove brain revealed that mast cell-derived material can enter neurons in three ways: by direct fusion of the granule and plasma membranes (mast cell and neuron); by capture of insoluble granule remnants and, potentially, via receptor-mediated endocytosis of gonadotropin-releasing hormone, a soluble mediator derived from the mast cell. These processes result in differential subcellular localization of mast cell material in neurons, including free in the neuronal cytoplasm, membrane bound in granule-like compartments or in association with small vesicles and the trans-Golgi network. Capture of granule remnants is the most frequently observed form of neuronal acquisition of mast cell products and correlates quantitatively with mast cells undergoing piecemeal degranulation. The present study indicates that mast cell-derived products can enter neurons, a process termed transgranulation, indicating a novel form of brain-immune system communication. PMID- 16262661 TI - Melatonin inhibits hippocampal long-term potentiation. AB - The goal of this study is to investigate the effect of the hormone melatonin on long-term potentiation and excitability measured by stimulating the Schaffer collaterals and recording the field excitatory postsynaptic potential from the CA1 dendritic layer in hippocampal brain slices from mice. Application of melatonin produced a concentration-dependent inhibition of the induction of long term potentiation, with a concentration of 100 nm producing an approximately 50% inhibition of long-term potentiation magnitude. Long-duration melatonin treatments of 6 h were also effective at reducing the magnitude of long-term potentiation. Melatonin (100 nm) did not alter baseline evoked responses or paired-pulse facilitation recorded at this synapse. The inhibitory actions of melatonin were prevented by application of the melatonin (MT) receptor antagonist luzindole as well as the MT2 receptor subtype antagonist 4-phenyl-2 propionamidotetraline. These inhibitory actions of melatonin were lost in mice deficient in MT2 receptors but not those deficient in MT1 receptors. In addition, application of the protein kinase A inhibitor H-89 both mimicked the effects of melatonin and precluded further inhibition by melatonin. Finally, the application an activator of adenylyl cyclase, forskolin, overcame the inhibitory effects of melatonin on LTP without affecting the induction of long-term potentiation on its own. These results suggest that hippocampal synaptic plasticity may be constrained by melatonin through a mechanism involving MT2-receptor-mediated regulation of the adenylyl cyclase-protein kinase A pathway. PMID- 16262664 TI - Hypotension and short-term anaesthesia induce ERK1/2 phosphorylation in autonomic nuclei of the brainstem. AB - The aims of this study were: first, to investigate the effects of anaesthesia on phosphorylated extracellular signal-regulated kinase (p-ERK)1/2-immunoreactivity (-ir) in the brainstem; second, to choose the best anaesthetic for p-ERK1/2 studies; and third, to determine the effect of short-term hypotension on p-ERK1/2 ir in the brainstem. Rats were anaesthetized with halothane, sodium pentobarbital or 100% CO2 narcosis, or were cervically dislocated and within 5 min perfused and the brains processed immunohistochemically for pERK1/2-ir. p-ERK1/2-ir was primarily observed in regions associated with cardiovascular and/or respiratory control. Several regions consistently showed dense p-ERK1/2 labelling, including a restricted region of the ventrolateral medulla (VLM). In contrast, other regions showed differential labelling depending on the mode of death. Cervical dislocation showed the least VLM labelling, limited to a discrete area approximately 0.6-1.4 mm caudal to the facial nucleus. Anaesthetics induced labelling throughout the VLM, with halothane inducing the most. Many p-ERK1/2-ir VLM neurons were catecholaminergic following halothane or sodium pentobarbital anaesthesia, but no double labelling was seen following cervical dislocation. Of the anaesthetics, sodium pentobarbital induced the least labelling and was used subsequently. Intravenous hydralazine was used to induce a 20-min period of hypotension, whereas arterial pressure did not change in vehicle-treated animals. Hydralazine evoked more pERK-ir neurons in specific regions, including the VLM, nucleus tractus solitarius (NTS), parabrachial nuclei, Kolliker-Fuse nucleus and locus coeruleus. Approximately twice as many p-ERK1/2-positive neurons were seen in the intermediate NTS and rostral VLM following hydralazine compared with the vehicle. In conclusion, p-ERK1/2-ir identifies neurons in central autonomic regions, and their number and distribution are markedly affected by anaesthetics, and are increased in some regions by short-term hypotension. PMID- 16262665 TI - Activity of individual reticulospinal neurons during different forms of locomotion in the lamprey. AB - Lamprey (a lower vertebrate) can employ different modes of locomotion, i.e. swimming in open water and crawling in tight places. Swimming is due to the periodic waves of lateral undulations with reciprocal activity of right and left muscles. In contrast, crawling (forward and backward) is based on single waves with coactivation of muscles on two sides. Basic mechanisms of swimming and, most likely, crawling reside in the spinal cord, and are activated by supraspinal commands. The main source of these commands is the reticulospinal (RS) system. The goal of the present experiments was to characterize the activity of individual RS neurons during swimming and during crawling in a U-shaped tunnel. The activity was recorded by means of chronically implanted electrodes in freely behaving animals. All recorded RS neurons were active during swimming but silent in quiescent animals. Many of them (61%) showed phasic modulation of their firing rate approximately in phase with the activity of ipsilateral rostral muscles. The majority of the neurons (80%) were also active during crawling. Many of them either increased or decreased their activity during crawling as compared to the background activity. These changes were better correlated with the direction of progression (forward or backward) than with the direction of turning in the tunnel (right or left). No correlation of the activity of RS neurons during locomotion and their sensory inputs was found. The results of this study suggest that different modes of locomotion in lampreys can be caused by considerably overlapping groups of RS neurons. PMID- 16262666 TI - Unilateral lesion of the pedunculopontine nucleus induces hyperactivity in the subthalamic nucleus and substantia nigra in the rat. AB - Recent data suggest a role for the pedunculopontine nucleus (PPN) in the pathophysiology of Parkinson's disease. Although there is anatomical evidence that the PPN and the basal ganglia are reciprocally connected, the functional importance of these connections is poorly understood. Lesioning of the PPN was shown to induce akinesia in primates, whereas in the 6-hydroxydopamine rat model the PPN was found to be hyperactive. As both nigrostriatal dopamine depletion and lesioning of the PPN were shown to induce akinesia and parkinsonism, the present study was performed in order to investigate the changes in neuronal activity of the subthalamic nucleus (STN) and the substantia nigra pars reticulata (SNr) after unilateral ibotenic acid lesioning of the PPN and after unilateral 6 hydroxydopamine lesioning of the substantia nigra pars compacta (SNc). The firing rate of STN neurones significantly increased from 10.2 +/- 6.2 (mean +/- SD) to 14.6 +/- 11.7 spikes/s after lesion of the PPN and to 18.6 +/- 14.5 spikes/s after lesion of the SNc. The activity of the SNr significantly increased from 19.6 +/- 10.5 to 28.7 +/- 13.4 spikes/s after PPN lesioning and to 23.5 +/- 10.8 spikes/s after SNc lesioning. Furthermore, PPN lesion decreased the number of spontaneously firing dopaminergic SNc cells, while having no effect on their firing rate. The results of our study show that lesion of the PPN leads to hyperactivity of the STN and SNr, similar to the changes induced by lesion of the SNc. Moreover, the decreased activity of SNc cells observed after PPN lesion might be at the origin of activity changes in the STN and SNr. PMID- 16262667 TI - Oxygen and glucose deprivation induces major dysfunction in the somatosensory cortex of the newborn rat. AB - The mechanisms and functional consequences of ischemia-induced injury during perinatal development are poorly understood. Subplate neurons (SPn) play a central role in early cortical development and a pathophysiological impairment of these neurons may have long-term detrimental effects on cortical function. The acute and long-term consequences of combined oxygen and glucose deprivation (OGD) were investigated in SPn and compared with OGD-induced dysfunction of immature layer V pyramidal cortical neurons (PCn) in somatosensory cortical slices from postnatal day (P)0-4 rats. OGD for 50 min followed by a 10-24-h period of normal oxygenation and glucose supply in vitro or in culture led to pronounced caspase-3 dependent apoptotic cell death in all cortical layers. Whole-cell patch-clamp recordings revealed that the majority of SPn and PCn responded to OGD with an initial long-lasting ischemic hyperpolarization accompanied by a decrease in input resistance (R(in)), followed by an ischemic depolarization (ID). Upon reoxygenation and glucose supply, the recovery of the membrane potential and R(in) was followed by a Na+/K+-ATPase-dependent postischemic hyperpolarization, and in almost half of the investigated SPn and PCn by a postischemic depolarization. Whereas neither a moderate (2.5 mm) nor a high (4.8 mm) increase in extracellular magnesium concentration protected the SPn from OGD-induced dysfunction, blockade of NMDA receptors with MK-801 led to a significant delay and decrease of the ID. Our data demonstrate that OGD induces apoptosis and a profound dysfunction in SPn and PCn, and underline the critical role of NMDA receptors in early ischemia-induced neuronal damage. PMID- 16262668 TI - Short-term constant light potentiation of large-magnitude circadian phase shifts induced by 8-OH-DPAT: effects on serotonin receptors and gene expression in the hamster suprachiasmatic nucleus. AB - Nonphotic phase-shifting of mammalian circadian rhythms is thought to be mediated in part by serotonin (5-HT) acting in the suprachiasmatic nucleus (SCN) circadian clock. Previously we showed that brief (1-3 days) exposure to constant light (LL) greatly potentiates nonphotic phase-shifting induced by the 5-HT agonist, (+/-)2 dipropyl-amino-8-hydroxyl-1,2,3,4-tetrahydronapthalene (8-OH-DPAT). Here we investigated potential mechanisms for this action of LL, including 5-HT receptor upregulation and SCN clock gene and neuropeptide gene expression. Autoradiographic analysis of ritanserin inhibition of [3H]8-OH-DPAT binding indicated that LL (approximately 2 days) did not affect 5-HT7 receptor binding in the SCN or dorsal raphe. Measurement of 5-HT1A autoreceptors in the median raphe and 5-HT1B receptors in the SCN also showed no effect of LL. In experiment 2, hamsters held under a 14-h light : 10-h dark photocycle (LD) or exposed to LL for approximately 2 days received an intraperitoneal injection of 8-OH-DPAT or vehicle at zeitgeber time (ZT) 6 or 0 and were killed after 2 h of dark exposure. 8-OH-DPAT suppressed SCN Per1 and Per2 mRNAs at both ZTs, as assessed by in situ hybridization. Per1 mRNA was also suppressed by LL alone. In addition, in situ hybridization of arginine vasopressin (AVP) mRNA and vasoactive intestinal polypeptide mRNA showed that LL significantly suppressed the former but not the latter. The LL-induced suppression of SCN Per1 mRNA and AVP mRNA may be involved in LL-induced potentiation of pacemaker resetting, especially as these data provide additional evidence that LL suppresses circadian pacemaker amplitude, thus rendering the clock more susceptible to phase-shifting stimuli. PMID- 16262670 TI - Tectal neurons signal impending collision of looming objects in the pigeon. AB - Although the optic tectum in non-mammals and its mammalian homolog, the superior colliculus, are involved in avoidance behaviors, whether and how tectal neurons respond to an object approaching on a collision course towards the animal remain unclear. Here we show by single unit recording that there exist three classes of looming-sensitive neurons in the pigeon tectal layer 13, which sends looming information to the nucleus rotundus or to the tectopontine system. The response onset time of tau cells is approximately constant whereas that for rho and eta cells depends on the square root of the diameter/velocity ratio of objects looming towards the animal, the cardioacceleration of which is also linearly related to the square root of this ratio. The receptive field of tectal cells is composed of an excitatory center and an inhibitory periphery, and this periphery does not inhibit responses to looming stimuli. These results suggest that three classes of tectal neurons are specified for detecting an object approaching on a collision course towards the animal, and that rho and eta cells may signal early warning of impending collision whereas tau cells initiate avoidance responses at a constant time before collision through the tectopontine system. PMID- 16262669 TI - Movement-related changes in oscillatory activity in the human subthalamic nucleus: ipsilateral vs. contralateral movements. AB - A voluntary movement is accompanied by a series of changes in neuronal oscillatory activity in the subthalamic nucleus (STN). These changes can be recorded through electrodes implanted for deep brain stimulation to treat Parkinson's disease in the time interval between the surgery and the internalization of the connections to the batteries. Both baseline activity and movement-related changes are different in the 'on' and 'off' medication motor states. In the 'off' state a low frequency activity in the alpha-beta range (8-25 Hz) that dominates the spectrum is interrupted during the movement, while in the 'on' state baseline frequencies are higher and a peri-movement gamma increase (70 80 Hz) is usually observed. Similar changes have been described with electrocorticographic recordings over the primary motor cortex but the gamma increase was only present during contralateral movements. We compared ipsi- and contralateral movement-related changes in STN activity, using a time-frequency analysis of the recordings obtained simultaneously in both STN and the scalp (electroencephalography) during right and left hand movements. The movement related changes observed in the STN in the 'on' and the 'off' states were similar to those described previously in terms of predominant frequency bands, but we found bilateral changes in the STN during movements of either hand. A contralateral earlier start of the beta STN changes was mostly observed when the moving hand corresponded to the less-affected side, irrespective of hand dominance. These results suggest that movement-related activity in the STN has, by and large, a bilateral representation and probably reflects cortical input. PMID- 16262671 TI - Morphine withdrawal-induced morphological changes in the nucleus accumbens. AB - Morphine withdrawal produces a hypofunction of mesencephalic dopamine neurons that impinge upon medium spiny neurons (MSN) of the forebrain. After chronic treatment (from 20 to 140 mg/kg of morphine twice a day over 14 days at escalating doses) rats were withdrawn from chronic morphine spontaneously and pharmacologically. In these two distinct conditions we studied the effects of withdrawal on the morphology of MSN of the core and shell of the nucleus accumbens (Nacc). MSN were stained with the Golgi-Cox procedure and analysed by a confocal laser-scanning microscope (CLSM). Our analysis shows that, shell and core MSN differed significantly for perikarya size and spine density, and the various morphine treatments did not affect the perikarya morphometry. Both spontaneous and naloxone-induced withdrawal produced a similar reduction in spine density in MS shell neurons, as compared with MS core neurons. This effect is selectively localized at the level of second order dendritic trunks where afferents converge. By contrast, spine density counts of accumbens MSN from rats chronically treated with morphine, did not reveal any change. Collectively, the results of the present study are twofold: (i) spontaneous and pharmacologically precipitated withdrawal, but not chronic morphine per se, affects spine density of target structures of a reduced mesolimbic dopamine transmission, and (ii) the reduction of spine density in second order dendritic trunks is selectively segregated in the MSN of the shell of the Nacc. In conclusion, morphine withdrawal dramatically alters spine density, selectively in second order dendritic trunks of Nacc shell MSN, thereby further impoverishing the already abated dopamine (DA) transmission. This is in line with recent views suggesting the hypodopaminergic state as a cardinal feature of opioid dependence. PMID- 16262672 TI - Opposing effects of amygdala and orbital prefrontal cortex lesions on the extinction of instrumental responding in macaque monkeys. AB - Extinction is a well-known behavioural phenomenon that allows organisms to respond flexibly to a changing environment. Although recent work implicates the amygdala and orbital prefrontal cortex (PFo) in extinction of Pavlovian conditioned fear and aversion, much less is known about the neural bases of instrumental extinction. To explore the contribution of the macaque amygdala to flexible responding in the face of changing reward contingency, we tested the effects of selective, excitotoxic lesions of the amygdala on extinction of an instrumental response. For comparison, we evaluated the effects of ablation of PFo on the same task. Amygdala lesions facilitated the extinction of instrumental responses, whereas lesions of PFo had the opposite effect. PMID- 16262674 TI - Segregation between acquisition and long-term memory in sensorimotor learning. AB - It is widely accepted that learning first involves generating new memories and then consolidating them into long-term memory. Thus learning is generally viewed as a single continuous process with two sequential stages; acquisition and consolidation. Here, we tested an alternative hypothesis proposing that acquisition and consolidation take place, at least partly, in parallel. Human subjects learned two visuomotor tasks. One task required moving a cursor under visuomotor rotation and the other required arbitrary association of colour to direction of movement. Subjects learned the two tasks in sequence, and were tested for acquisition of the second immediately after learning the first, and for retention of the first on the following day. The results show that learning one task led to proactive interference to acquisition of the second. However, this interference was not accompanied by retroactive interference to consolidation of the first task, indicating that acquisition and consolidation can be uncoupled. PMID- 16262673 TI - Differential involvement of the prelimbic cortex and striatum in conditioned heroin and sucrose seeking following long-term extinction. AB - Relapse to drug taking is triggered by stimuli previously associated with consumption of drugs of misuse (cues) and involves brain systems controlling motivated behaviour towards natural reinforcers. In this study, we aimed to identify and compare neuronal pathways in corticostriatal systems that control conditioned heroin or natural reward (sucrose) seeking. To that end, rats were trained to self-administer heroin or sucrose in association with an identical compound cue. After more than 3 weeks of abstinence during extinction training, cue exposure robustly reinstated heroin and sucrose seeking, but induced distinct and even opposing changes in the expression of the neuronal activation marker zif268 in the prelimbic cortex and striatal complex, respectively. Because in the prelimbic area zif268 expression was enhanced during cue-induced heroin seeking but unaffected during sucrose seeking, a pharmacological intervention was aimed at this prefrontal region. Injection of a GABA agonist mixture within the prelimbic area enhanced conditioned heroin seeking, but had no effect on conditioned sucrose seeking. Our findings suggest a differential role of the prelimbic area and the striatum in the persistence of heroin vs. sucrose seeking following long-term extinction. PMID- 16262675 TI - Home bases formed to visual cues but not to self-movement (dead reckoning) cues in exploring hippocampectomized rats. AB - Spatial theory proposes that the hippocampus contributes to exploratory behavior allowing animals to acquire information about their environment. In the present study, the exploratory movements of control rats, bulbectomized (anosmic) rats and hippocampectomized rats using the neurotoxin N-methyl-D-aspartate (NMDA) were monitored on a large circular table without walls and around which visual cues were manipulated. The rats displayed organized spatial behavior in that they developed home bases, one or more places operationally defined as those in which they spent a preponderance of time, in which they moved slowly, and to which they returned after excursions. Control rats and hippocampectomized rats were similar in that they established home bases: (i) adjacent to a proximal stable or moving visual landmark; (ii) in relation to more distant visual room cues; and (iii) in relation to contextually conditioned visual cues. Nevertheless, in exploratory tests given under infrared light, a wavelength to which rats are insensitive, control rats and bulbectomized rats established one or more home bases that were not dependent upon surface (e.g. olfactory) cues, whereas home base behavior was absent/fragmented in hippocampectomized rats. Thus, exploratory behavior, as exemplified by home base behavior, is organized in control and hippocampectomized rats in relation to visual cues, but is not organized in hippocampectomized rats when visual cues are absent. This result is discussed in relation to the idea that the hippocampus contributes to spatial behavior that is dependent upon guidance (dead reckoning) derived from self-movement cues. PMID- 16262676 TI - Memantine prolongs survival in an amyotrophic lateral sclerosis mouse model. AB - Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease which results from selective loss of upper and lower motor neurons. Mouse models of ALS, such as one carrying the G93A mutant of the human Cu-Zn superoxide dismutase gene[SOD1(G93A)], develop motor neuron pathology and clinical symptoms similar to those observed in ALS patients. There is compelling evidence that both direct and indirect glutamate toxicity contribute to the pathogenesis of motor neuron degeneration. However, the therapeutic effect of various glutamate receptor antagonists has not been clearly demonstrated. Memantine is a noncompetitive N methyl-D-aspartate (NMDA) receptor antagonist. It has been shown to protect neurons against NMDA- or glutamate-induced toxicity in vitro and in animal models of neurodegenerative diseases. In the current study, we have examined the therapeutic efficacy of memantine in an ALS mouse model carrying a high copy number of SOD1(G93A). Memantine treatment significantly delayed the disease progression and increased the life span of SOD1(G93A) mice, from 121.4 +/- 5.5 to 129.7 +/- 4.5 days (P = 0.032). Furthermore, NMDA receptor subunits were reliably detected in the spinal cord of SOD1(G93A) mice and their expression levels were similar to those in the wild-type littermate control. Therefore, the neuroprotective effect of memantine in SOD1(G93A) mice is most probably due to the inhibition of spinal cord NMDA receptors. In view of the long-term usage of memantine for dementia patients, with excellent tolerance and safety, these data suggest that memantine may be used in ALS patients alone or in combination with other therapies to prolong survival. PMID- 16262677 TI - A cytoplasmic motif targets neuroligin-1 exclusively to dendrites of cultured hippocampal neurons. AB - The formation of neuronal synapses is thought to depend on trans-synaptic interactions between cell adhesion molecules (CAMs) on the surface of axons and dendrites. Synapses are highly asymmetric structures. Pre- and post-synaptic domains might therefore be assembled around heterophilic CAMs which are polarized to axons vs. dendrites. We here investigated the targeting of neuroligin (NLG)-1, a heterophilic CAM, which promotes synapse formation through interaction with its receptor beta-neurexin in axons. We demonstrate that NLG-1 is highly polarized to the dendritic plasma membrane. Dendritic targeting relies on a cytoplasmic amino acid motif. By expressing chimeras of NLG-1 and CD8, an unpolarized protein, we show that the cytoplasmic domain of NLG-1 is necessary and sufficient for dendritic targeting. Furthermore, by truncation analysis we isolated a 32-amino acid targeting motif. When appended to CD8 this cytoplasmic sequence is sufficient to direct exclusively dendritic localization of the protein. Analysis of yellow fluorescent protein-tagged NLG-1 revealed that vesicular structures containing NLG-1 are excluded from the axon indicating that polarized distribution may be achieved by direct dendritic transport. We propose that the strict polarity of NLG-1 contributes to the directional assembly of synapses during development of the central nervous system. PMID- 16262678 TI - Species and strain differences in the expression of a novel glutamate-modulating cannabinoid receptor in the rodent hippocampus. AB - A novel, non-CB1 cannabinoid receptor has been defined by the persistence of inhibition of glutamatergic EPSPs by the cannabinoid receptor agonist WIN55,212-2 in mice lacking the cloned CB1 receptor (CB1-/-) (Hajos et al., 2001). This novel receptor was also distinguished from CB1 by its sensitivity to the antagonist SR141716A and its insensitivity to the antagonist AM251 (Hajos & Freund, 2002). We have chosen to refer to this putative receptor as CBsc due to its identification on Schaffer collateral axon terminals in the hippocampus. We examined properties of CBsc receptors in Sprague Dawley (SD) rats and two strains of wild-type (WT) mice (C57BL/6J and CD1) used as backgrounds for two independent lines of CB1-/- mice (Ledent et al., 1999; Zimmer et al., 1999). The inhibition of synaptic glutamate release by WIN55,212-2 was observed in hippocampal slices from WT CD1 mice and SD rats but was absent in WT C57 mice. We also found that AM251 and SR141716A antagonized the effect of WIN55,212-2 in hippocampal slices from CD1 mice and SD rats demonstrating a lack of selectivity of these ligands for CB1 and CBsc receptors in these animals. The results indicate that the glutamate-modulating CBsc cannabinoid receptor is present in the hippocampi of CD1 mice and SD rats but not in C57BL/6J mice. Thus, we have identified animal models that may permit the study of cannabinoids independently of the novel CBsc receptor (C57CB1+/+), the CBsc receptor independently of the cloned CB1 receptor (CD1CB1-/-), or in the absence of both receptors (C57CB1-/-). PMID- 16262679 TI - High-frequency repetitive transcranial magnetic stimulation over the hand area of the primary motor cortex disturbs predictive grip force scaling. AB - When we repetitively lift an object, our grip force is influenced by the mechanical object properties of the preceding lift, irrespective of whether the subsequent lift is performed with the same hand or the hand opposite to the preceding lift. This study investigates if repetitive high-frequency transcranial magnetic stimulation (rTMS) over the dominant primary motor cortex affects this relationship. After completion of 10 lifts of an object using the dominant hand, rTMS was applied over the dominant primary motor cortex for 20 s. On the first lift following rTMS, the peak grip force was significantly higher than on the lift preceding rTMS. Moreover, this measure remained elevated throughout the following set of lifts after rTMS. rTMS did not change the peak lift force generated by more proximal arm muscles. The same effect was observed when the lifts following rTMS over the dominant motor cortex were performed with the ipsilateral hand. These effects were not observed when subjects rested both hands on their lap or when a sham stimulation was applied for the same period of time. These preliminary data suggest that rTMS over the sensorimotor cortex disturbs predictive grip force planning. PMID- 16262682 TI - Advances in functional protein microarray technology. AB - Numerous innovations in high-throughput protein production and microarray surface technologies have enabled the development of addressable formats for proteins ordered at high spatial density. Protein array implementations have largely focused on antibody arrays for high-throughput protein profiling. However, it is also possible to construct arrays of full-length, functional proteins from a library of expression clones. The advent of protein-based microarrays allows the global observation of biochemical activities on an unprecedented scale, where hundreds or thousands of proteins can be simultaneously screened for protein protein, protein-nucleic acid, and small molecule interactions. This technology holds great potential for basic molecular biology research, disease marker identification, toxicological response profiling and pharmaceutical target screening. PMID- 16262681 TI - High-throughput two-hybrid analysis. The promise and the peril. AB - The two-hybrid method detects the interaction of two proteins by their ability to reconstitute the activity of a split transcription factor, thus allowing the use of a simple growth selection in yeast to identify new interactions. Since its introduction about 15 years ago, the assay largely has been applied to single proteins, successfully uncovering thousands of novel protein partners. In the last few years, however, two-hybrid experiments have been scaled up to focus on the entire complement of proteins found in an organism. Although a single such effort can itself result in thousands of interactions, the validity of these high throughput approaches has been questioned as a result of the prevalence of numerous false positives in these large data sets. Such artifacts may not be an obstacle to continued scale-up of the method, because the classification of true and false positives has proven to be a computational challenge that can be met by a growing number of creative strategies. Two examples are provided of this combination of high-throughput experimentation and computational analysis, focused on the interaction of Plasmodium falciparum proteins and of Saccharomyces cerevisiae membrane proteins. PMID- 16262683 TI - Emerging tools for real-time label-free detection of interactions on functional protein microarrays. AB - The availability of extensive genomic information and content has spawned an era of high-throughput screening that is generating large sets of functional genomic data. In particular, the need to understand the biochemical wiring within a cell has introduced novel approaches to map the intricate networks of biological interactions arising from the interactions of proteins. The current technologies for assaying protein interactions--yeast two-hybrid and immunoprecipitation with mass spectrometric detection--have met with considerable success. However, the parallel use of these approaches has identified only a small fraction of physiologically relevant interactions among proteins, neglecting all nonprotein interactions, such as with metabolites, lipids, DNA and small molecules. This highlights the need for further development of proteome scale technologies that enable the study of protein function. Here we discuss recent advances in high throughput technologies for displaying proteins on functional protein microarrays and the real-time label-free detection of interactions using probes of the local index of refraction, carbon nanotubes and nanowires, or microelectromechanical systems cantilevers. The combination of these technologies will facilitate the large-scale study of protein interactions with proteins as well as with other biomolecules. PMID- 16262685 TI - Mammalian initiator apoptotic caspases. AB - Caspases are a conserved family of cysteine proteases. They play diverse roles in inflammatory responses and apoptotic pathways. Among the caspases is a subgroup whose primary function is to initiate apoptosis. Within their long prodomains, caspases-2, -9 and -12 contain a caspase activation and recruitment domain while caspases-8 and -10 bear death effector domains. Activation follows the recruitment of the procaspase molecule via the prodomain to a high molecular mass complex. Despite sharing some common features, other aspects of the biochemistry, substrate specificity, regulation and signaling mechanisms differ between initiator apoptotic caspases. Defects in expression or activity of these caspases are related to certain pathological conditions including neurodegenerative disorders, autoimmune diseases and cancer. PMID- 16262684 TI - Examining multiprotein signaling complexes from all angles. AB - Dynamic protein-protein interactions are involved in most physiological processes and, in particular, for the formation of multiprotein signaling complexes at transmembrane receptors, adapter proteins and effector molecules. Because the unregulated induction of signaling complexes has substantial clinical relevance, the investigation of these complexes is an active area of research. These studies strive to answer questions about the composition and function of multiprotein signaling complexes, along with the molecular mechanisms of their formation. In this review, the adapter protein, linker for activation of T cells (LAT), will be employed as a model to exemplify how signaling complexes are characterized using a range of techniques. The intensive investigation of LAT highlights how the systematic use of complementary techniques leads to an integrated understanding of the formation, composition and function of multiprotein signaling complexes that occur at receptors, adapter proteins and effector molecules. PMID- 16262686 TI - Separation of a cholesterol-enriched microdomain involved in T-cell signal transduction. AB - We isolated a cholesterol-enriched membrane subpopulation from the so-called lipid raft fractions of Jurkat T-cells by taking advantage of its selective binding to a cholesterol-binding probe, BCtheta. The BCtheta-bound membrane subpopulation has a much higher cholesterol/phospholipid (C/P) molar ratio (approximately 1.0) than the BCtheta-unbound population in raft fractions (approximately 0.3). It contains not only the raft markers GM1 and flotillin, but also some T-cell receptor (TCR) signalling molecules, including Lck, Fyn and LAT. In addition, Csk and PAG, inhibitory molecules of the TCR signalling cascade, are also contained in the BCtheta-bound membranes. On the other hand, CD3epsilon, CD3zeta and Zap70 are localized in the BCtheta-unbound membranes, segregated from other TCR signalling molecules under nonstimulated conditions. However, upon stimulation of TCR, portions of CD3epsilon, CD3zeta and Zap70 are recruited to the BCtheta-bound membranes. The Triton X-100 concentration used for lipid raft preparation affects neither the C/P ratio nor protein composition of the BCtheta bound membranes. These results show that our method is useful for isolating a particular cholesterol-rich membrane domain of T-cells, which could be a core domain controlling the TCR signalling cascade. PMID- 16262688 TI - Structural evidence for a constant c11 ring stoichiometry in the sodium F-ATP synthase. AB - The Na+-dependent F-ATP synthases of Ilyobacter tartaricus and Propionigenium modestum contain membrane-embedded ring-shaped c subunit assemblies with a stoichiometry of 11. Subunit c from either organism was overexpressed in Escherichia coli using a plasmid containing the corresponding gene, extracted from the membrane using detergent and then purified. Subsequent analyses by SDS/PAGE revealed that only a minor portion of the c subunits had assembled into stable rings, while the majority migrated as monomers. The population of rings consisted mainly of c11, but more slowly migrating assemblies were also found, which might reflect other c ring stoichiometries. We show that they consisted of higher aggregates of homogeneous c11 rings and/or assemblies of c11 rings and single c monomers. Atomic force microscopy topographs of c rings reconstituted into lipid bilayers showed that the c ring assemblies had identical diameters and that stoichiometries throughout all rings resolved at high resolution. This finding did not depend on whether the rings were assembled into crystalline or densely packed assemblies. Most of these rings represented completely assembled undecameric complexes. Occasionally, rings lacking a few subunits or hosting additional subunits in their cavity were observed. The latter rings may represent the aggregates between c11 and c1, as observed by SDS/PAGE. Our results are congruent with a stable c11 ring stoichiometry that seems to not be influenced by the expression level of subunit c in the bacteria. PMID- 16262687 TI - Characterization and mode of action of an exopolygalacturonase from the hyperthermophilic bacterium Thermotoga maritima. AB - An intracellular pectinolytic enzyme, PelB (TM0437), from the hyperthermophilic bacterium Thermotoga maritima was functionally produced in Escherichia coli and purified to homogeneity. PelB belongs to family 28 of the glycoside hydrolases, consisting of pectin-hydrolysing enzymes. As one of the few bacterial exopolygalacturonases, it is able to remove monogalacturonate units from the nonreducing end of polygalacturonate. Detailed characterization of the enzyme showed that PelB is highly thermo-active and thermostable, with a melting temperature of 105 degrees C and a temperature optimum of 80 degrees C, the highest described to date for hydrolytic pectinases. PelB showed increasing activity on oligosaccharides with an increasing degree of polymerization. The highest activity was found on the pentamer (1000 U.mg(-1)). In addition, the affinity increased in conjunction with the length of the oligoGalpA chain. PelB displayed specificity for saturated oligoGalpA and was unable to degrade unsaturated or methyl-esterified oligoGalpA. Analogous to the exopolygalacturonase from Aspergillus tubingensis, it showed low activity with xylogalacturonan. Calculations on the subsite affinity revealed the presence of four subsites and a high affinity for GalpA at subsite +1, which is typical of exo-active enzymes. The physiological role of PelB and the previously characterized exopectate lyase PelA is discussed. PMID- 16262689 TI - Conformational studies of a hyperthermostable enzyme. AB - The structural features of the hyperthermophilic endo-beta-1,3-glucanase from Pyrococcus furiosus were studied using circular dichroism, steady-state and time resolved fluorescence spectroscopy and anisotropy. Upon heat and chemical treatment the folded and denatured states of the protein were characterized by distinguishable spectral profiles that identified a number of conformational states. The fluorescence methods showed that the spectral differences arose from changes in the local environment around specific tryptophan residues in the native, partially folded, partially unfolded and completely unfolded state. A structural resemblance was observed between the native protein and the structurally perturbed state which resulted after heat treatment at 110 degrees C. The enzyme underwent disruption of the native secondary and tertiary structure only after incubation at biologically extremely high temperatures (i.e. 150 degrees C), whilst in the presence of 8 m of guanidine hydrochloride the protein was partially unfolded. PMID- 16262690 TI - A new clan of CBM families based on bioinformatics of starch-binding domains from families CBM20 and CBM21. AB - Approximately 10% of amylolytic enzymes are able to bind and degrade raw starch. Usually a distinct domain, the starch-binding domain (SBD), is responsible for this property. These domains have been classified into families of carbohydrate binding modules (CBM). At present, there are six SBD families: CBM20, CBM21, CBM25, CBM26, CBM34, and CBM41. This work is concentrated on CBM20 and CBM21. The CBM20 module was believed to be located almost exclusively at the C-terminal end of various amylases. The CBM21 module was known as the N-terminally positioned SBD of Rhizopus glucoamylase. Nowadays many nonamylolytic proteins have been recognized as possessing sequence segments that exhibit similarities with the experimentally observed CBM20 and CBM21. These facts have stimulated interest in carrying out a rigorous bioinformatics analysis of the two CBM families. The present analysis showed that the original idea of the CBM20 module being at the C terminus and the CBM21 module at the N-terminus of a protein should be modified. Although the CBM20 functionally important tryptophans were found to be substituted in several cases, these aromatics and the regions around them belong to the best conserved parts of the CBM20 module. They were therefore used as templates for revealing the corresponding regions in the CBM21 family. Secondary structure prediction together with fold recognition indicated that the CBM21 module structure should be similar to that of CBM20. The evolutionary tree based on a common alignment of sequences of both modules showed that the CBM21 SBDs from alpha-amylases and glucoamylases are the closest relatives to the CBM20 counterparts, with the CBM20 modules from the glycoside hydrolase family GH13 amylopullulanases being possible candidates for the intermediate between the two CBM families. PMID- 16262691 TI - Spectroscopic and kinetic properties of the horseradish peroxidase mutant T171S. Evidence for selective effects on the reduced state of the enzyme. AB - Studies on horseradish peroxidase C and other haem peroxidases have been carried out on selected mutants in the distal haem cavity providing insight into the functional importance of the distal residues. Recent work has demonstrated that proximal structural features can also exert an important influence in determining the electronic structure of the haem pocket. To extend our understanding of the significance of proximal characteristics in regulating haem properties the proximal Thr171Ser mutant has been constructed. Thr171 is an important linking residue between the structural proximal Ca2+ ion and the proximal haem ligand, in particular the methyl group of Thr171 interdigitates with other proximal residues in the core of the enzyme. Although the mutation induces no significant changes to the functional properties of the enzyme, electronic absorption and resonance Raman spectroscopy reveal that it has a highly selective affect on the reduced state of the enzyme, effectively stabilizing it, whilst the electronic properties of the Fe(III) state unchanged and essentially identical to those of the native protein. This results in a significant change in the Fe2+/Fe3+ redox potential of the mutant. It is concluded that the unusual properties of the Thr171Ser mutant reflect the loss of a structural restraint in the proximal haem pocket that allows 'slippage' of the proximal haem ligand, but only in the reduced state. This is a remarkably subtle and specific effect that appears to increase the flexibility of the reduced state of the mutant compared to that of the wild-type protein. PMID- 16262692 TI - Uptake of bilirubin into HepG2 cells assayed by thermal lens spectroscopy. Function of bilitranslocase. AB - Bilitranslocase is a carrier protein localized at the basolateral domain of the hepatocyte plasma membrane. It transports various organic anions, including bromosulfophthalein and anthocyanins. Functional studies in subcellular fractions enriched in plasma membrane revealed a high-affinity binding site for bilirubin, associated with bilitranslocase. The aim of this work was to test whether the liver uptake of bilirubin depends on the activity of bilitranslocase. To this purpose, an assay of bilirubin uptake into HepG2 cell cultures was set up. The transport assay medium contained bilirubin at a concentration of approximately 50 nm in the absence of albumin. To analyse the relative changes in bilirubin concentration in the medium throughout the uptake experiment, a highly sensitive thermal lens spectrometry method was used. The mechanism of bilirubin uptake into HepG2 cells was investigated by using inhibitors such as anti-sequence bilitranslocase antibodies, the protein-modifying reagent phenylmethanesulfonyl fluoride and diverse organic anions, including nicotinic acid, taurocholate and digoxin. To validate the assay further, both bromosulfophthalein and indocyanine green uptake in HepG2 cells was also characterized. The results obtained show that bilitranslocase is a carrier with specificity for both bilirubin and bromosulfophthalein, but not for indocyanine green. PMID- 16262693 TI - Structures of the O-polysaccharides and classification of Proteus genomospecies 4, 5 and 6 into respective Proteus serogroups. AB - An acidic branched O-polysaccharide was isolated by mild acid degradation of the lipopolysaccharide (LPS) of Proteus genomospecies 4 and studied by sugar and methylation analyses along with 1H and 13C NMR spectroscopy, including 2D COSY, TOCSY, ROESY and H-detected 1H, 13C HSQC experiments. The following structure of the pentasaccharide repeating unit of the O-polysaccharide was established, which is unique among Proteus polysaccharide structures: [structure: see text] where Qui3NAc stands for 3-acetamido-3,6-dideoxyglucose. Based on the O-polysaccharide structure and serological data, we propose classifying Proteus genomospecies 4 into a new, separate Proteus serogroup, O56. A weak cross-reactivity of Proteus genomospecies 4 antiserum with LPS of Providencia stuartii O18 and Proteus vulgaris OX2 was observed and is discussed in view of a similarity of the O polysaccharide structures. Structural and serological investigations showed that Proteus genomospecies 5 and 6 should be classified into the existing Proteus serogroups O8 and O69, respectively. PMID- 16262694 TI - In vitro gamma-secretase cleavage of the Alzheimer's amyloid precursor protein correlates to a subset of presenilin complexes and is inhibited by zinc. AB - The gamma-secretase complex mediates the final proteolytic event in Alzheimer's disease amyloid-beta biogenesis. This membrane complex of presenilin, anterior pharynx defective, nicastrin, and presenilin enhancer-2 cleaves the C-terminal 99 amino acid fragment of the amyloid precursor protein intramembranously at gamma sites to form C-terminally heterogeneous amyloid-beta and cleaves at an epsilon site to release the intracellular domain or epsilon-C-terminal fragment. In this work, two novel in vitro gamma-secretase assays are developed to further explore the biochemical characteristics of gamma-secretase activity. During development of a bacterial expression system for a substrate based on the amyloid precursor protein C-terminal 99-amino acid sequence, fragments similar to amyloid-beta and an epsilon-C-terminal fragment were observed. Upon purification this substrate was used in parallel with a transfected source of substrate to measure gamma secretase activity from detergent extracted membranes. With these systems, it was determined that recovery of size-fractionated cellular and tissue-derived gamma secretase activity is dependent upon detergent concentration and that activity correlates to a subset of high molecular mass presenilin complexes. We also show that by changing the solvent environment with dimethyl sulfoxide, detection of epsilon-C-terminal fragments can be elevated. Lastly, we show that zinc causes an increase in the apparent molecular mass of an amyloid precursor protein gamma secretase substrate and inhibits its cleavage. These studies further refine our knowledge of the complexes and biochemical factors needed for gamma-secretase activity and suggest a mechanism by which zinc dysregulation may contribute to Alzheimer's disease pathogenesis. PMID- 16262695 TI - The chloroplast ClpP complex in Chlamydomonas reinhardtii contains an unusual high molecular mass subunit with a large apical domain. AB - The composition of the chloroplast-localized protease complex, ClpP, from the green alga Chlamydomonas reinhardtii was characterized by nondenaturing electrophoresis, immunoblotting and MS. The detected ClpP complex has a native mass of approximately 540 kDa, which is approximately 200 kDa higher than ClpP complexes in higher plant chloroplasts, mitochondria or bacteria. The 540-kDa ClpP complex contains two nuclear-encoded ClpP proteins (ClpP3 and P5) and five ClpR (R1, R2, R3, R4 and R6) proteins, as well two proteins, ClpP1L and ClpP1H, both probably derived from the plastid clpP1 gene. ClpP1H is 59 kDa and contains a approximately 30-kDa insertion sequence (IS1) not found in other ClpP proteins, responsible for the high MW of the complex. Based on comparison with other sequences, IS1 protrudes as an additional domain on the apical surface of the ClpP/R complex, probably preventing interaction with the HSP100 chaperone. ClpP1L is a 25-kDa protein similar in size to other ClpP proteins and could arise by post-translational processing of ClpP1H. Chloramphenicol-chase experiments show that ClpP1L and ClpP1H have a similar half-life, indicating that both are stable components of the complex. The structure of the ClpP complex is further discussed in conjunction with a phylogenetic analysis of the ClpP/R genes. A model is proposed for the evolution of the algal and plant complex from its cyanobacterial ancestor. PMID- 16262696 TI - Atrial natriuretic peptide-dependent photolabeling of a regulatory ATP-binding site on the natriuretic peptide receptor-A. AB - The natriuretic peptide receptor-A (NPR-A) is composed of an extracellular ligand binding domain, a transmembrane-spanning domain, a kinase homology domain (KHD) and a guanylyl cyclase domain. Because the presence of ATP or adenylylimidodiphosphate reduces atrial natriuretic peptide (ANP) binding and is required for maximal guanylyl cyclase activity, a direct interaction of ATP with the receptor KHD domain is plausible. Therefore, we investigated whether ATP interacts directly with a binding site on the receptor by analyzing the binding of a photoaffinity analog of ATP to membranes from human embryonic kidney 293 cells expressing the NPR-A receptor lacking the guanylyl cyclase moiety (DeltaGC). We demonstrate that this receptor (NPR-A-DeltaGC) can be directly labeled by 8-azido-3'-biotinyl-ATP and that labeling is highly increased following ANP treatment. The mutant receptor DeltaKC, which does not contain the KHD, is not labeled. Photoaffinity labeling of the NPR-A-DeltaGC is reduced by 50% in the presence of 550 microm ATP, and competition curve fitting studies indicate a Hill slope of 2.2, suggestive of cooperative binding. This approach demonstrates directly that the interaction of ANP with its receptor modulates the binding of ATP to the KHD, probably through a conformational change in the KHD. In turn, this conformational change is essential for maximal activity. In addition, the ATP analog, 8-azido-adenylylimidodiphosphate, inhibits guanylyl cyclase activity but increases ANP binding to the extracellular domain. These results suggest that the KHD regulates ANP binding and guanylyl cyclase activity independently. PMID- 16262697 TI - Biochemical characterization of human umbilical vein endothelial cell membrane bound acetylcholinesterase. AB - Acetylcholinesterase is an enzyme whose best-known function is to hydrolyze the neurotransmitter acetylcholine. Acetylcholinesterase is expressed in several noncholinergic tissues. Accordingly, we report for the first time the identification of acetylcholinesterase in human umbilical cord vein endothelial cells. Here we further performed an electrophoretic and biochemical characterization of this enzyme, using protein extracts obtained by solubilization of human endothelial cell membranes with Triton X-100. These extracts were analyzed under polyacrylamide gel electrophoresis in the presence of Triton X-100 and under nondenaturing conditions, followed by specific staining for cholinesterase or acetylcholinesterase activity. The gels revealed one enzymatically active acetylcholinesterase band in the extracts that disappeared when staining was performed in the presence of eserine (an acetylcholinesterase inhibitor). Performing western blotting with the C-terminal anti acetylcholinesterase IgG, we identified a single protein band of approximately 70 kDa, the molecular mass characteristic of the human monomeric form of acetylcholinesterase. The western blotting with the N-terminal anti acetylcholinesterase IgG antibody revealed a double band around 66-70 kDa. Using the Ellman's method to measure the cholinesterase activity in human umbilical vein endothelial cells, regarding its substrate specificity, we confirmed the existence of an acetylcholinesterase enzyme. Our studies revealed a predominance of acetylcholinesterase over other cholinesterases in human endothelial cells. In conclusion, we have demonstrated the existence of a membrane-bound acetylcholinesterase in human endothelial cells. In future studies, we will investigate the role of this protein in the endothelial vascular system. PMID- 16262698 TI - Secretion of the mammalian Sec14p-like phosphoinositide-binding p45 protein. AB - Protein-lipid interactions are important for protein targeting, signal transduction, lipid transport, and the maintenance of cellular compartments and membranes. Specific lipid-binding protein domains, such as PH, FYVE, PX, PHD, C2 and SEC14 homology domains, mediate interactions between proteins and specific phospholipids. We recently cloned a 45-kDa protein from rat olfactory epithelium, which is homologous to the yeast Sec14p phosphatidylinositol (PtdIns) transfer protein and we report here that this protein binds to PtdIns(3,4,5)P3 and far weaker to less phosphorylated derivatives of PtdIns. Expression of the p45 protein in COS-1 cells resulted in accumulation of the protein in secretory vesicles and in the extracellular space. The secreted material contained PtdIns(3,4,5)P3. Our findings are the first report of a Sec14p-like protein involved in transport out of a cell and, to the best of our knowledge, inositol containing phospholipids have not previously been detected in the extracellular space. Our findings suggest that p45 and phosphoinositides may participate in the formation of the protective mucus on nasal epithelium. PMID- 16262699 TI - Characterization of solanesyl and decaprenyl diphosphate synthases in mice and humans. AB - The isoprenoid chain of ubiquinone (Q) is determined by trans-polyprenyl diphosphate synthase in micro-organisms and presumably in mammals. Because mice and humans produce Q9 and Q10, they are expected to possess solanesyl and decaprenyl diphosphate synthases as the determining enzyme for a type of ubiquinone. Here we show that murine and human solanesyl and decaprenyl diphosphate synthases are heterotetramers composed of newly characterized hDPS1 (mSPS1) and hDLP1 (mDLP1), which have been identified as orthologs of Schizosaccharomyces pombe Dps1 and Dlp1, respectively. Whereas hDPS1 or mSPS1 can complement the S. pombe dps1 disruptant, neither hDLP1 nor mDLP1 could complement the S. pombe dLp1 disruptant. Thus, only hDPS1 and mSPS1 are functional orthologs of SpDps1. Escherichia coli was engineered to express murine and human SpDps1 and/or SpDlp1 homologs and their ubiquinone types were determined. Whereas transformants expressing a single component produced only Q8 of E. coli origin, double transformants expressing mSPS1 and mDLP1 or hDPS1 and hDLP1 produced Q9 or Q10, respectively, and an in vitro activity of solanesyl or decaprenyl diphosphate synthase was verified. The complex size of the human and murine long chain trans-prenyl diphosphate synthases, as estimated by gel-filtration chromatography, indicates that they consist of heterotetramers. Expression in E. coli of heterologous combinations, namely, mSPS1 and hDLP1 or hDPS1 and mDLP1, generated both Q9 and Q10, indicating both components are involved in determining the ubiquinone side chain. Thus, we identified the components of the enzymes that determine the side chain of ubiquinone in mammals and they resembles the S. pombe, but not plant or Saccharomyces cerevisiae, type of enzyme. PMID- 16262701 TI - Response of the Pacific oyster Crassostrea gigas to hypoxia exposure under experimental conditions. AB - The molecular response to hypoxia stress in aquatic invertebrates remains relatively unknown. In this study, we investigated the response of the Pacific oyster Crassostrea gigas to hypoxia under experimental conditions and focused on the analysis of the differential expression patterns of specific genes associated with hypoxia response. A suppression subtractive hybridization method was used to identify specific hypoxia up- and downregulated genes, in gills, mantle and digestive gland, after 7-10 days and 24 days of exposure. This method revealed 616 different sequences corresponding to 12 major physiological functions. The expression of eight potentially regulated genes was analysed by RT-PCR in different tissues at different sampling times over the time course of hypoxia. These genes are implicated in different physiological pathways such as respiration (carbonic anhydrase), carbohydrate metabolism (glycogen phosphorylase), lipid metabolism (delta-9 desaturase), oxidative metabolism and the immune system (glutathione peroxidase), protein regulation (BTF3, transcription factor), nucleic acid regulation (myc homologue), metal sequestration (putative metallothionein) and stress response (heat shock protein 70). Stress proteins (metallothioneins and heat shock proteins) were also quantified. This study contributes to the characterization of many potential genetic markers that could be used in future environmental monitoring, and could lead to explore new mechanisms of stress tolerance in marine mollusc species. PMID- 16262700 TI - Cdk2 activity is dispensable for triggering replicon initiation after transient hypoxia in T24 cells. AB - We examined whether the fast release of replicon initiation after sudden O2 recovery of hypoxically incubated mammalian cells depends on kinase activity of Cdk2. We used a system based on starved/refed T24 cells elaborated previously for such investigations [van Betteraey-Nikoleit M, Eisele KH, Stabenow D and Probst H (2003) Eur J Biochem270, 3880-3890]. Cells subjected to hypoxia concurrently with refeeding accumulate the G1 DNA content within 5-6 h. In this state they are ready to perform, within 1-2 min after O2 recovery, a burst of replicon initiations that marks the start of a synchronous S-phase. We found that Cdk2 binds to the chromatin fraction within 4-6 h after refeeding with fresh medium, irrespective of whether the cells were incubated normoxically or hypoxically. However, inhibition of Cdk2 by olomoucine, roscovitine or the Cdk2/cyclin inhibitory peptide II had no influence on the synchronous burst of replicon initiations. Cdc6 and pRb, possible targets of Cdk2 phosphorylation, behaved differentially. Inhibition did not affect phosphorylation of Cdc6 after reoxygenation, whilst chromatin bound pRb remained hypophosphorylated beyond the initiation burst. Thus, neither Cdk2 activity, though present at the end of the hypoxic period, nor pRb phosphorylation are necessary for releasing the burst of replicon initiations upon oxygen recovery. Consequentially, Cdk2 dependent phosphorylation(s) cannot be a critical trigger of replicon initiation in response to reoxygenation after several hours of hypoxia, at least in the T24 cells studied. PMID- 16262702 TI - Identification and biochemical characterization of the Anopheles gambiae 3 hydroxykynurenine transaminase. AB - Spontaneous oxidation of 3-hydroxykynureine (3-HK), a metabolic intermediate of the tryptophan degradation pathway, elicits a remarkable oxidative stress response in animal tissues. In the yellow fever mosquito Aedes aegypti the excess of this toxic metabolic intermediate is efficiently removed by a specific 3-HK transaminase, which converts 3-HK into the more stable compound xanthurenic acid. In anopheline mosquitoes transmitting malaria, xanthurenic acid plays an important role in Plasmodium gametocyte maturation and fertility. Using the sequence information provided by the Anopheles gambiae genome and available ESTs, we adopted a PCR-based approach to isolate a 3-HK transaminase coding sequence from the main human malaria vector A. gambiae. Tissue and developmental expression analysis revealed an almost ubiquitary profile, which is in agreement with the physiological role of the enzyme in mosquito development and 3-HK detoxification. A high yield procedure for the expression and purification of a fully active recombinant version of the protein has been developed. Recombinant A. gambiae 3-HK transaminase is a dimeric pyridoxal 5'-phosphate dependent enzyme, showing an optimum pH of 7.8 and a comparable catalytic efficiency for both 3-HK and its immediate catabolic precursor kynurenine. This study may be useful for the identification of 3-HK transaminase inhibitors of potential interest as malaria transmission-blocking drugs or effective insecticides. PMID- 16262703 TI - Analysis of the stability of the spermadhesin PSP-I/PSP-II heterodimer. Effects of Zn2+ and acidic pH. AB - Spermadhesins are a family of 12-16 kDa proteins with a single CUB domain. PSP-I and PSP-II, the most abundant boar spermadhesins, are present in seminal plasma as a noncovalent heterodimer. Dimerization markedly affects the binding ability of the subunits. Notably, heparin and mannose 6-phosphate binding abilities of PSP-II are abolished, indicating that the corresponding binding sites may be located at (or near) the dimer interface. Pursuing the hypothesis that cryptic binding sites in PSP-I/PSP-II may be exposed in specific physiological environments, we examined the influence of Zn2+ and acidic pH on the heterodimer stability. According to near-UV CD spectra, the core native fold is preserved in the presence of physiological concentrations of Zn2+, a cation unusually abundant in boar seminal plasma. However, the thermostability of the heterodimer decreases significantly, as observed by CD and differential scanning calorimetry. The effect is Zn2+-specific and is reversed by EDTA. Destabilization is also observed at acidic pH. Gel filtration analysis using radioiodinated PSP-I/PSP-II reveals that dissociation of the heterodimer at low (nanomolar) protein concentrations is promoted by both Zn2+ and acidic pH. Although the integrity of the heterodimer in seminal plasma seems to be guaranteed by its high concentration, dissociation may be facilitated in the female genital tract because of dilution of the protein in the intraluminal fluids of the cervix and the uterus, and the acidic fluid of the uterotubal junction. Such a mechanism may be relevant in the regulation of uterine immune reactions. PMID- 16262704 TI - DNA hypomethylation reduces homologous pairing of inserted tandem repeat arrays in somatic nuclei of Arabidopsis thaliana. AB - Fluorescent chromatin tagging makes possible tracking of specific loci in vivo and in situ. Loci tagged by the lac operator (lacO)/GFP-LacI/Nuclear Localization Signal (NLS) system show rapid motility and constrained chromatin dynamics in somatic nuclei of a transgenic line, designated EL702C, in Arabidopsis thaliana. The tagged loci associated with each other significantly more often than expected at random, due to homologous pairing of the lacO tandem repeat arrays. Furthermore, these arrays associated significantly more often than average euchromatic regions with heterochromatic chromocenters (CCs). We show now that the inserted lacO array in this transgenic line became strongly methylated at CG sites in the T3 generation, which can be reversed upon transfer into the mutant backgrounds of decrease in DNA methylation 1 (ddm1) and methyltransferase 1 (met1). Concomitantly, the tagged loci showed lower association frequencies as compared with the transgenics in wild-type background, which is correlated with a significant decrease in allelic and ectopic pairing of the lacO repeat arrays as visualized by fluorescence in situ hybridization. In contrast, the preferential association of the lacO arrays with heterochromatin, locus mobility in somatic nuclei and transcription of neighboring transgenes were not altered by reduced DNA methylation in ddm1 and met1 backgrounds. Our results show that repeat arrays can activate hypermethylation of the inserted locus that correlates with high frequencies of homologous pairing in somatic cells. In contrast, the preferential association of these inserted arrays with CCs in plant cells occurs through another mechanism. PMID- 16262705 TI - Consistent transcriptional silencing of 35S-driven transgenes in gentian. AB - In this study, no transgenic gentian (Gentiana triflora x Gentiana scabra) plants produced via Agrobacterium-mediated transformation exhibited transgene (GtMADS, gentian-derived MADS-box genes or sGFP, green fluorescent protein) expression in their leaf tissues, despite the use of constitutive Cauliflower mosaic virus (CaMV) 35S promoter. Strikingly, no expression of the selectable marker gene (bar) used for bialaphos selection was observed. To investigate the possible cause of this drastic transgene silencing, methylation-specific sequences were analysed by bisulfite genomic sequencing using tobacco transformants as a control. Highly methylated cytosine residues of CpG and CpWpG (W contains A or T) sites were distinctively detected in the promoter and 5' coding regions of the transgenes 35S-bar and 35S-GtMADS in all gentian lines analysed. These lines also exhibited various degrees of cytosine methylation in asymmetrical sequences. The methylation frequencies in the other transgene, nopaline synthase (NOS) promoter driven nptII, and the endogenous GtMADS gene coding region, were much lower and were variable compared with those in the 35S promoter regions. Transgene methylation was observed in the bialaphos-selected transgenic calluses expressing the transgenes, and methylation sequences were distributed preferentially around the as-1 element in the 35S promoter. Calluses derived from leaf tissues of silenced transgenic gentian also exhibited transgene suppression, but expression was recovered by treatment with the methylation inhibitor 5-aza-2'-deoxycytidine (aza-dC). These results indicated that cytosine methylation occurs exclusively in the 35S promoter regions of the expressed transgenes during selection of gentian transformants, causing transcriptional gene silencing. PMID- 16262706 TI - Analysis of the histone H3 gene family in Arabidopsis and identification of the male-gamete-specific variant AtMGH3. AB - Histones are major components of chromatin, the protein-DNA complex involved in DNA packaging and transcriptional regulation. Histone genes have been extensively investigated at the genome level in animal systems and have been classified as replication dependent, replication independent or tissue specific. However, no such study is available in a plant system. In this paper we report that there are 15 histone H3 genes in the Arabidopsis genome, including five H3.1 genes, three H3.3 genes and five H3.3-like genes. A gene structure analysis revealed that gene duplication causes redundancy of the histone H3 genes. The expression of one of the H3 genes, termed AtMGH3/At1g19890, is cell-specific, being restricted to the generative and sperm cells of Arabidopsis pollen as shown by in situ hybridisation and reporter gene analysis. Thus, we conclude that in Arabidopsis, AtMGH3 is a male-gamete-specific histone H3 gene. A T-DNA insertion line for AtMGH3 revealed decreased expression and ectopic RNA splicing. The T-DNA insertion lines for AtMGH3/At1g19890 and other H3 genes revealed a normal growth phenotype and reproductive fertility. These findings suggest that other H3 genes are likely to compensate for the T-DNA-insertion-induced loss of a single H3 gene because of the high redundancy of these genes in the Arabidopsis genome. These T DNA mutant lines should be useful for accumulating different H3 gene mutations in a single plant and for studying replication-dependent and replication-independent H3 genes and the specific role of AtMGH3 in chromatin remodelling and transcriptional regulation during development of male gametes. PMID- 16262707 TI - Hormonally controlled expression of the Arabidopsis MAX4 shoot branching regulatory gene. AB - The Arabidopsis MORE AXILLARY BRANCHING 4 (MAX4) gene is required for the production of a long-range, graft-transmissible signal that inhibits shoot branching. Buds of max4 mutant plants are resistant to the inhibitory effects of apically applied auxin, indicating that MAX4 is required for auxin-mediated bud inhibition. The RAMOSUS 1 (RMS1) and DECREASED APICAL DOMINANCE 1 (DAD1) genes of pea and petunia, respectively, are orthologous to MAX4 and function in a similar way. Here we show that, despite the similarities between these three genes, there are significant differences in the regulation of their expression. RMS1 is known to be upregulated by auxin in the shoot, suggesting a straightforward link between the RMS1-dependent branch-inhibiting signal and auxin, whereas we find that MAX4 is only upregulated by auxin in the root and hypocotyl, and this is not required for the inhibition of shoot branching. Furthermore, both RMS1 and DAD1 are subject to feedback regulation, for which there is no evidence for MAX4. Instead, overexpression studies and reciprocal grafting experiments demonstrate that the most functionally significant point of interaction between auxin and MAX4 is post-transcriptional and indeed post-synthesis of the MAX4-dependent graft-transmissible signal. PMID- 16262708 TI - Localization of tabersonine 16-hydroxylase and 16-OH tabersonine-16-O methyltransferase to leaf epidermal cells defines them as a major site of precursor biosynthesis in the vindoline pathway in Catharanthus roseus. AB - The Madagascar periwinkle (Catharanthus roseus) produces the well known and remarkably complex anticancer dimeric alkaloids vinblastine and vincristine, which are derived by the coupling of vindoline and catharanthine monomers. Recent data from in situ RNA hybridization and immunolocalization suggest that combinatorial cell factories within the leaf are involved in vindoline biosynthesis. In this study, the cell types responsible for vindoline biosynthesis were identified by laser-capture microdissection/RNA isolation/RT PCR to show that geraniol hydroxylase, secologanin synthase, tryptophan decarboxylase, strictosidine synthase, strictosidine ss-glucosidase and tabersonine 16-hydroxylase can be detected preferentially in epidermal cells. A new and complementary application of the carborundum abrasion (CA) technique was developed to obtain epidermis-enriched leaf extracts that can be used to measure alkaloid metabolite levels, enzyme activities and gene expression. The CA technique showed that tabersonine and 16-methoxytabersonine, together with 16 hydroxytabersonine-16-O-methyltransferase, are found predominantly in Catharanthus leaf epidermis, in contrast to vindoline, catharanthine and later enzymatic steps in vindoline biosynthesis. The results show that leaf epidermal cells are biosynthetically competent to produce tryptamine and secologanin precursors that are converted via many enzymatic transformations to make 16 methoxytabersonine. This alkaloid or its 2,3 dihydro-derivative is then transported to cells (mesophyll/idioblast/laticifer) within Catharanthus leaves to complete the last three or four enzymatic transformations to make vindoline. PMID- 16262709 TI - Appearance of actin microfilament 'twin peaks' in mitosis and their function in cell plate formation, as visualized in tobacco BY-2 cells expressing GFP-fimbrin. AB - The actin cytoskeleton of higher plants plays an essential role in plant morphogenesis and in maintaining various cellular activities. In this study we established a tobacco BY-2 cell line, stably transformed with a GFP-fimbrin actin binding domain (ABD) 2 construct, that allows visualization of actin microfilaments (MFs) in living cells. Using this cell line, designated BY-GF11, we performed time-sequential observations of MF dynamics during cell-cycle progression. Detailed analyses revealed the appearance of a broad MF band in the late G2 phase that separated to form a structure corresponding to the so-called actin-depleted zone (ADZ) in mitosis. In BY-GF11, the MF structure at the cell cortex in mitosis appeared to form two bands rather than the ADZ. Measurements of fluorescent intensities of the cell cortex indicated an MF distribution that resembled two peaks, and we therefore named the structure MF 'twin peaks' (MFTP). The cell plate formed exactly within the valley between the MFTP at cytokinesis, and this cell-plate guidance was distorted by disruption of the MFTP by an inhibitor of actin polymerization. These results suggest that the MFTP originates from the broad MF band in the G2 phase and functions as a marker of cytokinesis. PMID- 16262710 TI - The biosynthesis and regulation of biosynthesis of Concord grape fruit esters, including 'foxy' methylanthranilate. AB - The biosynthesis of methyl anthranilate, the volatile compound responsible for the distinctive 'foxy' aroma and flavor of the Washington Concord grape (Vitis labrusca), involves an alcohol acyltransferase that catalyzes the formation of methyl anthranilate from anthraniloyl-coenzyme A (CoA) and methanol. Although methanol is a poor substrate in comparison with the co-substrate, high levels of this acyltransferase (0.5% of the total protein) combined with relatively high levels of this alcohol make this reaction possible in grapes. This 449 amino acid protein belongs to the BAHD family of acyltransferases, having 58% identity with the benzoyl CoA:benzyl alcohol benzoyl transferase from Clarkia. Both native and recombinant enzymes can use a broad range of acyl-CoAs and alcohols as substrates. The ability of Concord grape alcohol acyltransferase to accept a range of different CoA esters and alcohols suggests this to be a versatile ester forming enzyme, similar to those of other fruits that than can produce a range of fruit esters based on the supply of appropriate substrates. Expression is coordinately regulated, with transcript, protein and enzyme activities coinciding with the accumulation of methyl anthranilate that occurs after the initiation of berry ripening. The majority of acyltransferase protein in grape tissues is localized to the outer fruit mesocarp, a result consistent with the fact that methyl anthranilate is released to the external environment throughout the ripening process. Wine grapes (Vitis vinifera) that accumulate neither anthranilate nor methyl anthranilate do not express this enzyme activity nor do they accumulate this protein. PMID- 16262711 TI - Identification of a plastid acyl-acyl carrier protein synthetase in Arabidopsis and its role in the activation and elongation of exogenous fatty acids. AB - Plant cells are known to elongate exogenously provided fatty acid (FA), but the subcellular sites and mechanisms for this process are not currently understood. When Arabidopsis leaves were incubated with 14C-FAs with or=20 carbons) but not synthesis of 14C-unsaturated 18-carbon or 16-carbon FAs. Isolated pea chloroplasts were also able to elongate 14C-FAs (300 mumol/l] and 24 infants with a history of NS were identified from hospital records. These children were compared to 40 children from the community (CC) without neonatal problems. At ages 18-32 months, the children's neurological, motor and developmental status were assessed, and blood groups of the NJ and NS subjects and their mothers were determined. Ten (43%) of the NJ subjects were unable to sit and/or stand independently. The NJ subjects had significantly more neurological, motor and developmental difficulties and caused greater maternal concern than the CCs. Five (21%) of the NJ subjects had possible blood group incompatibility. The NS subjects had significantly more motor and eye hand difficulties and maternal concerns expressed than the CCs. Severe NJ in term infants (of mainly non-haemolytic origin) was associated with a high prevalence of neurological and developmental sequelae at ages 18-32 months. The NS is also associated with neuro-developmental sequelae, but the pattern is different to those seen in NJ. Since NS is common in resource poor countries, this may be an important cause of neuro-developmental impairment in children living in this setting. PMID- 16262737 TI - Do growth monitoring and promotion programs answer the performance criteria of a screening program? A critical analysis based on a systematic review. AB - OBJECTIVE: Growth Monitoring and Promotion programs (GMP) have been intensively promoted to improve children's health in developing countries. It has been hoped that regularly weighing children would result in the early detection of growth falterers, and that the growth chart would serve as an educational tool to make that state apparent to both health workers and caretakers in order to trigger improved caring practices. Our objective was to review whether GMP answers the theoretical grounds of a screening and intervention program. METHOD: A systematic literature review was performed. The WHO framework developed by Wilson and Jungner for planning and evaluating screening programs guided the analysis. RESULTS: Sixty-nine studies were retrieved. Overall, evidence is weak on the performance of GMP as a screening program for malnutrition through early detection of growth falterers. The main results are: (1) malnutrition remains a public health problem, but its importance is context specific; (2) the value of a low weight velocity to predict malnutrition is unknown and likely to vary in different contexts; (3) the performance of GMP for improving nutrition status of children and in reducing mortality and morbidity is unknown; (4) the performance of the screening is affected by the unreliability of weight measurements; (5) the promotional and educational effectiveness of GMP is low, in particular the growth chart is poorly understood by mothers; (6) the acceptability seems low in regards of low attendance rates; (7) evidence is lacking regarding cost-effectiveness. CONCLUSIONS: We conclude that there is too little scientific evidence to indiscriminately support international promotion of GMP. However GMP could constitute a valid strategy of public nutrition in specific situations. We indicate paths for further research and how prevention programs could be developed. PMID- 16262738 TI - Use of intermittent preventive treatment for malaria in pregnancy in a rural area of western Kenya with high coverage of insecticide-treated bed nets. AB - Kenya established intermittent preventive treatment (IPT) with sulfadoxine pyrimethamine (SP) for malaria in pregnancy as national policy in 1998. We assessed the coverage of IPT among women who had recently delivered in a rural area of western Kenya with perennial malaria transmission and high coverage with insecticide treated nets (ITNs) through a cross-sectional, community-based survey in December 2002. Antenatal clinic (ANC) attendance was high (89.9% of the 635 participating women); 77.5% of attendees visited an ANC before the third trimester and 91.9% made more than one visit. Delivery of SP by the ANC was reported by 19.1% of all women but only 6.8% reported receiving more than one dose. Given the high rate of use of ANC services, if SP were given at each visit after the first trimester, the potential coverage of IPT (two doses of SP) would be 80.3% in this study population. ITNs were used by 82.4% of women during pregnancy, and almost all mothers (98.5%) who slept under an ITN shared the nets with their newborns after delivery. Women who thought malaria in pregnancy caused foetal problems were more likely to have used an ITN (adjusted odds ratio [AOR] 1.6, 95% confidence interval [CI] 1.0-2.4), and to have visited ANC more than once (AOR 2.4, 95% CI 1.2-4.7) compared to women who thought malaria in pregnancy was either not a problem or caused problems for the mother only. These findings illustrate the need for improved IPT coverage in this rural area. Identification and removal of the barriers to provision of IPT during ANC visits can help to increase coverage. In this area of Kenya, health messages stressing that foetal complications of malaria in pregnancy may occur in the absence of maternal illness may improve the demand for IPT. PMID- 16262739 TI - Evaluation of long-lasting insecticidal nets after 2 years of household use. AB - Development of long-lasting insecticidal nets (LLINs) may eliminate the need for insecticide retreatment of ITNs. While two LLINs (Olyset, Sumitomo Chemical Co., Japan; and PermaNet 1.0, Vestergaard-Frandsen, Denmark) have received recommendations from the World Health Organization Pesticide Evaluation Scheme, field-testing under normal use has been limited. We used a survival analysis approach to compare time to net failure of conventional polyester bednets treated only with deltamethrin to two LLINs and two candidate LLINs (Olyset; PermaNet; Insector, Athanor, France; and Dawa, Siamdutch Mosquito Netting Co., Thailand). Additionally, we evaluated nets treated with a process designed to increase the wash-durability of permethrin-treated nets through the addition of cyclodextrin (a starch) in the treatment process. Houses in western Kenya were randomly assigned to one of the six net types and nets were distributed to cover all sleeping spaces. Households were visited monthly to assess reported side effects in inhabitants and washing frequency. Nets were evaluated for insecticidal activity by periodic WHO cone bioassays with mortality assessed at 24 h. Nets with bioassay mortality <70% were assayed monthly until failure, defined as the first of two consecutive bioassay mortality rates <50%. Time to failure was analyzed using an extended Cox Proportional Hazards model controlling for the cumulative number of washes. We distributed 314 nets to 177 households in June July 2002; 22 nets (7.0%) were lost to follow-up and 196 (62.4%) failed during the first 2 years of the evaluation. Controlling for cumulative number of washes, PermaNet 1.0 [Hazard Ratio (HR) 0.14, 95% Confidence Interval (CI) 0.06-0.31] had a significantly lower risk of failure than conventional nets while Insector had a significantly higher risk of failure (HR 2.57, 95% CI 1.06-4.15). The risks of failure of the remaining nets (Olyset: HR 1.29, 95% CI 0.79-2.10; Dawa: HR 0.58, 95% CI 0.32-1.18; cyclodextrin: HR 0.65, 95% CI 0.40-1.1) were not significantly different from that of a conventional net. PermaNet 1.0 performed significantly better than conventional nets and should be recommended to malaria control programs. PMID- 16262740 TI - Integrating insecticide-treated bednets into a measles vaccination campaign achieves high, rapid and equitable coverage with direct and voucher-based methods. AB - Population coverage of insecticide-treated nets (ITNs) in Africa falls well below the Abuja target of 60% while coverage levels achieved during vaccination campaigns in the same populations typically exceed 90%. Household (HH) cost of ITNs is an important barrier to their uptake. We investigated the coverage, equity and cost of linking distribution of free ITNs to a measles vaccination campaign. During a national measles vaccination campaign in Zambia, children in four rural districts were given a free ITN when they received their measles vaccination. In one urban district, children were given a voucher, which could be redeemed for a net at a commercial distribution site. About 1700 HHs were asked whether they received vaccination and an ITN during a measles campaign, as well as questions on assets (e.g. type roofing material or bicycle ownership) to assess HH wealth. Net ownership was calculated for children in each wealth quintile. In the rural areas, ITN coverage among children rose from 16.7% to 81.1% and the equity ratio from 0.32 to 0.88 and in the urban area from 50.7% to 76.2% (equity ratio: 0.66-1.19). The operational cost per ITN delivered was dollar 0.35 in the rural area with direct distribution and $1.89 in the urban areas with voucher distribution. Mass distribution of ITNs through vaccination campaigns achieves rapid, high and equitable coverage at low cost. PMID- 16262742 TI - A review of the quality of randomized clinical trials of adjunctive therapy for the treatment of cerebral malaria. AB - BACKGROUND: Randomized controlled clinical trials (RCTs) of adjunctive treatment to reduce the high-mortality associated with cerebral malaria (CM) have so far failed to show any benefit. This may be due in part to improperly designed and/or conducted trials. Therefore a systematic review of quality of RCTs for the treatment of CM with mortality as either primary or secondary outcome published between 1980 and 2000, was conducted. METHODS: RCTs from the peer-reviewed literature using electronic searches. Methodological quality was assessed using an individual component approach (adequacy of concealment of allocation schedule, generation of allocation sequence, double blinding and analysis of participant as randomized). Sample sizes were recalculated for the ability of reviewed trials to detect 25% and 50% reductions in mortality. RESULTS: Nine trials satisfied the inclusion criteria and were reviewed. Only two had sufficient power to detect a 50% reduction in mortality, and none could detect a 25% reduction. All the trials had inadequate methodological quality in one or more of the components, although in two trials these deficiencies were few. CONCLUSION: There is a need for researchers and donors to ensure proper planning and implementation of RCTs in developing countries. In CM, demonstration of worthwhile reduction in mortality by a single intervention will require a large number of subjects, which a single centre may not be able to recruit. PMID- 16262741 TI - Open randomized study of artesunate-amodiaquine vs. chloroquine-pyrimethamine sulfadoxine for the treatment of uncomplicated Plasmodium falciparum malaria in Nigerian children. AB - BACKGROUND: Artemisinin-based combination antimalarials are currently considered effective alternatives for the treatment of malaria in Africa, but there are few studies of such combinations in Nigerian children. We assessed the safety, treatment efficacy and effects on gametocyte carriage of the combination of artesunate plus amodiaquine and chloroquine plus pyrimethamine-sulfadoxine in children. METHODS: We evaluated 153 children who were aged 12 years or younger who had uncomplicated Plasmodium falciparum malaria. Patients were randomly assigned a combination of artesunate (4 mg/kg of body weight daily for 3 days) plus amodiaquine (30 mg/kg over 3 days), or chloroquine (25 mg/kg over 3 days) plus pyrimethamine-sulfadoxine (25 mg/kg of the sulfadoxine component at presentation). The primary endpoints were the proportions of children with adequate clinical and parasitological response, late parasitological failure, late clinical failure and early treatment failure. The parasitological cure rates on days 14-28 were also used as the primary endpoints. RESULTS: Both regimens were well tolerated; no child was withdrawn because of drug intolerance. All children treated with artesunate plus amodiaquine had adequate clinical and parasitological response (ACPR), while all but five children treated with chloroquine plus pyrimethamine-sulfadoxine had similar response. Fever clearance times were similar in the two treatment groups. However, the proportion of patients whose parasitaemia cleared by day 2 was significantly higher (100 vs. 50%, P = 0.00001) and parasite clearance was significantly faster (1.7 +/- 0.4 vs. 2.5 +/- 0.8 days, P = 0.0001) in children treated with artesunate plus amodiaquine. The cure rates on days 21 (100%vs. 94%, P = 0.03) and 28 (100%vs. 90%, P = 0.003) were also significantly higher in children treated with artesunate plus amodiaquine than in those treated with chloroquine plus pyrimethamine-sulfadoxine. Overall, a significantly higher proportion of children treated with chloroquine plus pyrimethamine-sulfadoxine carried gametocytes at least once during follow-up compared with those treated with artesunate plus amodiaquine [5 of 50 (10%) vs. 1 of 103 (0.97%), P = 0.01]. CONCLUSION: The combination of artesunate plus amodiaquine is therapeutically superior to a combination of chloroquine plus pyrimethamine-sulfadoxine, and significantly reduced gametocyte carriage following treatment. PMID- 16262743 TI - Mutations in Plasmodium falciparum dihydrofolate reductase and dihydropteroate synthase genes in Senegal. AB - Senegal recently (2004) switched to sulfadoxine-pyrimethamine (SP) with amodiaquine as first line therapy for malaria in response to increasing chloroquine resistance. In anticipation of emerging resistance to SP as a result of this change in drug pressure, we set out to define the baseline prevalence of SP-associated mutations in the dhfr and dhps genes in Plasmodium falciparum using geographically diverse and longitudinally collected samples. A total of 153 blood samples were analysed from patients (5 years or older) with mild malaria after informed consent was obtained. Longitudinal samples were collected between 2000 and 2003 in Pikine, a suburb of Dakar. Geographically diverse site sampling was carried out in 2003. The mutation prevalence in DHFR codons 51, 59 and 108 is 65%, 61% and 78% in Pikine, 2003. The overall prevalence of the triple mutation that is associated with high-level pyrimethamine resistance is 61%. The mutation prevalence rate in DHPS codons 436 and 437 is 21% and 40%, respectively. There is significant geographic variation in genotypic resistance, as samples from Pikine in 2003 had higher mutation prevalence in the pfdhfr and pfdhps genes compared to samples from Tambacounda (P < 0.015). In summary, this study demonstrates a high background prevalence of SP resistance mutations already present in P. falciparum in Senegal. PMID- 16262744 TI - Hypothalamic-pituitary gonadal axis and immune response imbalance during chronic filarial infections. AB - Bi-directional relationships operate between the hypothalamic-pituitary-gonadal axis and the immune system. Cytokines, peptide hormones and their shared receptors/ligands are used as a common biological language for communication within and between the immune and neuroendocrine systems. Such communication suggests an immunoregulatory role for the brain and a sensory function for the immune system. We used a radioimmunoassay to measure the concentrations of steroid hormones (cortisol, testosterone, estradiol and progesterone) and pituitary hormones [follicle stimulating hormone (FSH), luteinizing hormone (LH) human chorionic gonadotropin (HCG) and prolactin] in peripheral blood plasma from 78 young Gabonese women with chronic filarial infections. We used an enzyme linked immunosorbent assay to determine the concentrations of four proinflammatory cytokines [tumor necrosis factor-alpha (TNF-alpha), gamma interferon (IFN-gamma), interleukin-1 (IL-1) and IL-6] in the same plasma samples. Progesterone was unchanged and all other steroid hormone plasma concentrations were lower in microfilaremic women than in amicrofilaremic women. The concentration of LH was higher in amicrofilaremic women, whereas the prolactin concentration was higher in microfilaremics. The plasma concentrations of TNF-alpha, IFN-gamma, IL-1 and IL-6 were higher in microfilaremic women. A strong negative correlation was found between the steroid and pituitary hormones and the pro-inflammatory cytokines. Conversely, a strong positive correlation was found between prolactin and the same cytokines. These data provide first evidence of immune system and hormonal system disturbance during chronic filarial infections and suggest that the observed imbalance should be taken into account in the diagnosis and treatment of filarial infections. PMID- 16262745 TI - Soil-transmitted helminthiasis in Uganda: epidemiology and cost of control. AB - A country-wide description of the distribution of soil-transmitted helminths in Uganda is reported, based on data for 20-185 school-children from 271 schools. The overall prevalence of Ascaris lumbricoides, Trichuris trichiura and hookworm was 6.3%, 5.0% and 43.5%, respectively. The prevalence of A. lumbricoides and T. trichiura was unevenly distributed in the country with prevalence greatest in south-western Uganda whereas hookworm was generally more homogeneously distributed. Based on preliminary cost analysis of an ongoing school-based control programme, the financial delivery cost per school-child treated with albendazole is estimated to be between US dollar 0.04 and 0.08 in different districts. PMID- 16262746 TI - Repellent properties of celery, Apium graveolens L., compared with commercial repellents, against mosquitoes under laboratory and field conditions. AB - In our search for new bioactive products against mosquito vectors, we reported the slightly larvicidal and adulticidal potency, but remarkable repellency of Apium graveolens both in laboratory and field conditions. Repellency of the ethanolic preparation of hexane-extracted A. graveolens was, therefore, investigated and compared with those of 15 commercial mosquito repellents including the most widely used, DEET. Hexane-extracted A. graveolens showed a significant degree of repellency in a dose-dependent manner with vanillin added. Ethanolic A. graveolens formulations (10-25% with and without vanillin) provided 2-5 h protection against female Aedes aegypti. Repellency that derived from the most effective repellent, 25% of hexane-extracted A. graveolens with the addition of 5% vanillin, was comparable to the value obtained from 25% of DEET with 5% vanillin added. Moreover, commercial repellents, except formulations of DEET, showed lower repellency than that of A. graveolens extract. When applied on human skin under field conditions, the hexane-extracted A. graveolens plus 5% vanillin showed a strong repellent action against a wide range of mosquito species belonging to various genera. It had a protective effect against Aedes gardnerii, Aedes lineatopennis, Anopheles barbirostris, Armigeres subalbatus, Culex tritaeniorhynchus, Culex gelidus, Culex vishnui group and Mansonia uniformis. The hexane-extracted A. graveolens did not cause a burning sensation or dermal irritation when applied to human skin. No adverse effects were observed on the skin or other parts of the human volunteers' body during 6 months of the study period or in the following 3 months, after which time observations ceased. Therefore, A. graveolens can be a potential candidate for use in the development of commercial repellents that may be an alternative to conventional synthetic chemicals, particularly in community vector control applications. PMID- 16262748 TI - Attributable fraction of malaria in Mali by Dicko et al. (2005). PMID- 16262749 TI - Tibor J. Greenwalt; in memoriam. PMID- 16262747 TI - Post-surgical assessment of excised tissue from patients with Buruli ulcer disease: progression of infection in macroscopically healthy tissue. AB - OBJECTIVE: The current standard of treatment of Buruli ulcer disease (BUD) is surgical excision of lesions. Excision size is determined macroscopically assuming the complete removal of all infected tissue. However, dissemination of infection beyond the excision margins into apparently healthy tissue, possibly associated with recurrences, cannot be excluded in this way. To assess the central to peripheral progression of Mycobacterium ulcerans infection and the mycobacterial infiltration of excision margins, excised tissue was examined for signs of infection. METHODS: 20 BUD lesions were excised in general anaesthesia including all necrotic and subcutaneous adipose tissue down to the fascia and at an average of 40 mm into the macroscopically unaffected tissue beyond the border of the lesion. Tissue samples were subjected to PCR and histopathology. RESULTS: Although the bacillary load decreased from central to peripheral, M. ulcerans infection was detected throughout all examined tissue specimens including the peripheral segments as well as excision margins of all patients. During the post operative hospitalization period (averaging 2 months) no local recurrences were observed. CONCLUSION: Available data suggest a correlation of surgical techniques with local recurrences. The results of this study indicate the unnoticed early progression of mycobacterial infection into macroscopically healthy tissue. Thus, the removal of all infected tissue cannot always be verified visually by the surgeon. Provided that long-term follow up of patients with positive excision margins will establish the clinical relevance of these findings, on-site laboratory assessment of excised tissue in combination with follow up may contribute to reduce recurrence rates. PMID- 16262750 TI - Variant Creutzfeldt-Jakob disease transmission by plasma products: assessing and communicating risk in an era of scientific uncertainty. AB - A substantial body of animal data indicates that transmissible spongiform encephalopathies (TSEs) are transmitted through blood. These data have been augmented in the past year by reports that two recipients of red cells from donors with variant Creutzfeldt-Jakob disease (vCJD) in the United Kingdom have acquired this infection. Most of the blood donations collected in countries affected by bovine spongiform encephalopathy (BSE) and vCJD also contribute plasma to fractionation pools. Thus, a number of batches of fractionated products have included plasma from donors who developed vCJD. On the basis of public health strategies influenced, in part, by risk assessments, the UK and the French authorities have instituted measures for recalling products and informing patients of the estimated risks. It is therefore relevant to review the principles used by authorities in generating risk assessments for the transmission of TSEs by blood and blood products. While the general principles are fairly straightforward, the final assessments are very dependent on the magnitude of several key parameters, which are, largely, still unknown. A critical determinant of final product risk is the extent to which the plasma fractionation process will contribute to eliminating the infectious prion agent. Therefore, regulatory and industry measures to characterize fractionation processes for their capacity to eliminate prions are to be strongly encouraged. In the interim, an understanding of the principles used to generate risk assessments should contribute to an enhanced ability to address this threat to patient safety. Authorities should recognize that adequate communication is an integral part of good risk-management practices. PMID- 16262751 TI - Evaluation of COBAS AmpliPrep nucleic acid extraction in conjunction with COBAS AmpliScreen HBV DNA, HCV RNA and HIV-1 RNA amplification and detection. AB - BACKGROUND AND OBJECTIVES: This report describes the evaluation of the COBAS AmpliPrep instrument for fully automated generic nucleic acid extraction in conjunction with hepatitis B virus (HBV) DNA, hepatitis C virus (HCV) RNA, and human immunodeficiency virus (HIV)-1 RNA COBAS AmpliScreen amplification and detection using serial dilutions of the WHO international standards (IS) and the PeliCheck reference panels. MATERIALS AND METHODS: Serial diluted samples of the WHO IS and the PeliCheck reference panels were tested 24 times to determine the HBV DNA, HCV RNA and HIV-1 RNA detection limits by Probit analysis. The existence and extent of cross-contamination were assessed by testing alternating high titre HBV DNA-positive and -negative samples. The specificity of the AmpliPrep AmpliScreen test for HBV was determined by testing 232 minipools consisting of six donations, all negative for HCV/HIV-1 nucleic acid testing (NAT) and HBsAg. In addition, a HBV genotypes A-G panel was tested. RESULTS: The respective 95% detection limits (and 95% CI) on the WHO IS and on the PeliCheck reference panels were 6.7 (4.3-13) IU/ml and 123 (68-301) gEq/ml for HBV DNA, 23 (11-106) IU/ml and 126 (84-233) gEq/ml for HCV RNA, and 187 (108-422) IU/ml and 183 (108-434) gEq/ml for HIV-1 RNA. Based on the WHO IS and the PeliCheck reference panels, no significant differences in sensitivity for HBV and HCV were found between AmpliPrep and the licensed MultiPrep extraction method. The sensitivity of AmpliPrep-AmpliScreen for HIV-1 was probably twofold lower as compared to the MultiPrep-AmpliScreen method. No cross contamination was observed. All 232 minipools were HBV NAT-negative. The AmpliPrep-AmpliScreen test for HBV detected HBV genotypes A-G with equal sensitivity. CONCLUSIONS: The AmpliPrep instrument combined with the AmpliScreen assays for HBV, HCV and HIV-1 is robust and suitable for NAT donor screening. The sensitivity criteria for HIV-1 and HCV as defined by the Paul Ehrlich Institute and the Food and Drug Administration for minipool NAT screening are met by this system. SINGLE SENTENCE SUMMARY: Generic COBAS AmpliPrep nucleic acid extraction in conjunction with COBAS AmpliScreen detection for HBV, HCV and HIV-1. PMID- 16262752 TI - Performance of ORTHO HCV core antigen and trak-C assays for detection of viraemia in pre-seroconversion plasma and whole blood donors. AB - OBJECTIVE: Logistics and cost of nucleic acid amplification testing (NAT) screening preclude its current use in many developing countries. Development of hepatitis C virus (HCV) core antigen assays offer an alternative to NAT. We evaluated two specimen populations to assess the sensitivity, relative to NAT, of the HCV core antigen (HCVcAg) ELISA (enzyme-linked immunosorbent assay) test system and the trak-C assay: (1) plasma donor HCV NAT-conversion panels and (2) cross-sectional whole blood donor NAT yield specimens. METHODS: Differential sensitivities among NAT (NGI; Chiron/Gen-Probe) and both HCVcAg assays (Ortho Clinical Diagnostics, Rochester, NY) were evaluated using: (1) 102 serial ramp-up phase specimens from 37 plasma donor NAT-conversion panels (Alpha Therapeutic/BioClinical Partners); and (2) 42 cross-sectional whole blood donor NAT yield specimens (confirmed RNA positive, antibody negative) plus 54 NAT false positive specimens (American Red Cross). RESULTS: Viral load among the plasma donor NAT-conversion panels at the cutoffs for HCVcAg and trak-C assays were 32 000 copies/ml (95% confidence interval [CI] 8000-120 000) and 8000 copies/ml (95% CI: 2200-28 000), respectively. The mean (95% CI) difference in window period reduction compared to routine mini-pool NAT screening (estimated sensitivity 100 copies/ml) was delayed 5.2 days (2.2-7.6 days) for HCVcAg assay and 3.8 days (2.1 5.5 days) for the trak-C assay. Among the 42 NAT yield specimens, the HCVcAg assay detected 31 (74%) as core antigen-positive while the trak-C assay detected 37 (88%) as core antigen-positive. Viral loads for the five specimens not detected by the trak-C HCVcAg assay ranged from 100 to 7770 copies/ml. All 54 NAT false-positive specimens were non-reactive on both HCV core antigen assays. CONCLUSION: These data indicate that the trak-C assay has sensitivity approaching routine mini-pool NAT screening for the detection of seronegative HCV infection. In the absence of routine NAT screening for early HCV infection, the use of an HCV core antigen assay should be considered. PMID- 16262753 TI - Optimal sampling time after preparation of platelet concentrates for detection of bacterial contamination by quantitative real-time polymerase chain reaction. AB - BACKGROUND AND OBJECTIVES: A universal quantitative real-time polymerase chain reaction (PCR), based on bacterial 16S rDNA, to detect bacterial contamination of platelet concentrates (PCs), was developed previously and compared with automated culturing. In the present study, this real-time PCR method was evaluated to determine the optimal sampling time for screening of bacterial contamination in PCs. MATERIALS AND METHODS: Routinely prepared PCs were spiked with suspensions of Escherichia coli, Bacillus cereus, Staphylococcus epidermidis, Pseudomonas aeruginosa and Propionibacterium acnes to 1, 10 and 100 colony-forming units (CFU)/ml and stored at room temperature for 7 days. The presence of bacteria in these PCs was monitored by quantitative real-time PCR. As a reference method (additional control), BacT/Alert automated culturing was used. For PCR, 1-ml aliquots were drawn from all (spiked) PCs on days 0, 1, 2, 3, 6 and 7 of storage. As a control, triplicate samples (10 ml) were inoculated into aerobic and anaerobic BacT/Alert culture bottles immediately after spiking (day 0) and after storage for 1, 2, 3, 6 or 7 days. RESULTS: With quantitative real-time PCR, all bacterial species tested were reproducibly detected on day 1 after spiking at original concentrations of 10 and 100 CFU/ml. Bacteria were also detected on day 1 from PCs spiked with an initial concentration of 1 CFU/ml, except for E. coli, which was detected in only one of the three samples and P. aeruginosa, for which analysis was not performed on day 1. With the reference method, bacteria were detected in culture bottles (inoculated on day 0) within a mean time of 20.1 h, with the exception of P. acnes which was detected at a mean time of 102.3 and 49.3 h (for original spiking concentrations of 10 and 100 CFU/ml respectively). CONCLUSIONS: PCR enables the rapid detection of low initial numbers of bacteria in PCs. For reliable detection, our results support that sampling of PCs for real time PCR screening should not be carried out earlier than 1 day after preparation (48 h after blood collection). Importantly, the real-time PCR approach has the potential to be used before the release of PCs from the blood centre or shortly before they are transfused in the hospital. PMID- 16262754 TI - Comparison of X-ray vs. gamma irradiation of CPDA-1 red cells. AB - BACKGROUND AND OBJECTIVES: Transfusion-associated graft-versus-host disease (TA GVHD) is a serious, potential complication of blood transfusion that is essentially prevented by blood product irradiation. Blood product irradiation is currently performed using gamma irradiation. X-ray irradiation is an alternative that has certain advantages: an X-ray irradiator is less expensive and does not have a radioactive source. However, the biochemical effects of X-ray irradiation on red blood cells (RBCs) have not been well characterized. The primary purpose of our study was to compare the effects of X-ray irradiation with gamma irradiation on RBC membrane permeability. A secondary purpose was to verify that X-ray irradiation has the same effect on lymphocytes as gamma irradiation. MATERIALS AND METHODS: Ten RBC units were split into two portions; five units of each were then irradiated with either gamma or X-ray at a dose of 35 Gy or 25 Gy, respectively. Free plasma haemoglobin and extracellular potassium levels were measured in each unit over time. Another unit was divided into three parts for lymphocyte studies; one part was not irradiated as a control and the other two received 25 Gy of irradiation via X-ray or gamma irradiation, respectively. RESULTS: Minimally increased free plasma haemoglobin was found only in X-ray irradiated units after 25 Gy compared to gamma-irradiated units. However, extracellular potassium levels were not significantly different in these groups. Extracellular potassium levels were slightly higher only in gamma irradiated units at 35 Gy. X-ray and gamma irradiation were equivalent in affecting lymphocyte function. CONCLUSIONS: Small differences in RBC membrane permeability are found between gamma-irradiated and X-ray-irradiated units. However, these differences are not likely to be clinically important. PMID- 16262755 TI - In-vitro evaluation of the PALL Leukotrap Affinity Prion Reduction Filter as a secondary device following primary leucoreduction. AB - BACKGROUND AND OBJECTIVES: A filter (PRF1) designed to remove abnormal prion proteins from red-cell units has been developed. The purpose of this study was to evaluate the quality of red cells produced using this device. MATERIALS AND METHODS: Leucocyte-depleted red-cell units (CPD, CPD-A1 and CPD/SAGM) processed according to standard UK practices were filtered using PRF1. Filtered and control units were stored and sampled on day 1, day 7 and on the date of expiry and were tested using standard measures of red-cell quality. RESULTS: Filtered units were found to have significantly higher percentage haemolysis levels, lower haemoglobin levels and a smaller volume compared with controls. All results, however, were well within the permitted 0.8% haemolysis level at the end of storage and all units met the UK guidelines for haemoglobin and volume. The other test parameters measured showed no significant differences between the test and control units. CONCLUSIONS: The PRF1 filter was found to be easy to use and resulted in red-cell units that met all relevant UK and European Guidelines. PMID- 16262756 TI - Factors associated with granulocyte colony-stimulating factor-induced peripheral blood stem cell yield in healthy donors. AB - BACKGROUND AND OBJECTIVES: Poor collection results are a clinical problem in granulocyte-colony stimulating factor (G-CSF)-induced peripheral blood stem cell (PBSC) collection in healthy donors. It would be beneficial to be able to predict the PBSC yield from allogeneic donors before mobilization or harvesting. MATERIALS AND METHODS: We examined the relationship between certain donor characteristics and the effectiveness of G-CSF-induced PBSC collection in 59 healthy family donors aged 3-63 years old (median 16 years). G-CSF was administered subcutaneously at 10 microg/kg for mobilization, daily for 5 days, and PBSC harvest using a continuous blood cell separator was started on day 5 of G-CSF treatment. Total cell yields were calculated as the number per unit of processed blood (l) per unit weight of the donor (kg). RESULTS: In a univariate analysis, the donor's age, body mass index (BMI), white blood cell (WBC) count before mobilization, and platelet count before and during mobilization were significantly correlated with the yield of mononuclear cells (MNC), CD34(+) cells and granulocyte-macrophage colony-forming units (GM-CFU). Younger age (P < 0.001), a low BMI (P = 0.002), a high WBC count before mobilization (P = 0.004), a high platelet count before (P = 0.012) and during (P < 0.05) mobilization, and a low speed of withdrawal (P = 0.019) were associated with a higher CD34(+) cell yield. No significant correlation was found for gender, the type of G-CSF, the serum level of G-CSF, the type of cell separator, or the type of blood access. A multivariate forward and backward stepwise selection regression analysis showed that the factors associated with CD34(+) cell yield were age, platelet count before and during mobilization, and circulating CD34(+) cell concentration on day 2 of G-CSF treatment. CONCLUSION: In this small preliminary study, we found that donor age is the most important factor in predicting G-CSF-induced PBSC yields. Old age and low platelet counts before mobilization might be useful indicators for identifying poor mobilizers. Further validation of these findings in a larger number of donors are needed to establish whether these findings apply to other populations. PMID- 16262757 TI - Motivation, recruitment and retention of voluntary non-remunerated blood donors: a survey-based questionnaire study. AB - BACKGROUND AND OBJECTIVES: The aim of this study was to establish which motivational and socio-demographic factors are important for the development of a long-term commitment as a voluntary, non-remunerated blood donor. STUDY DESIGN AND METHODS: A cross-sectional sample survey of active blood donors in Oslo, Norway, was conducted. Donors filled in a self-administered questionnaire during donation. Data on motivation were analysed using factor analysis. RESULTS: The blood donors' socio-demographic characteristics were found to be similar to those of the population as a whole. The single, most important, recruitment channel was the influence of active blood donors. Five dimensions of blood-donor motivation were identified with factor analysis. These were: altruism and empathy; social reasons (such as the influence of friends and family); strengthening of one's self-esteem; positive experiences associated with donation; and a moral obligation to donate. Support for statements on altruistic motives for donation was strong and similar in long-time and short-time donors. In contrast, short time donors were more likely to be motivated by factors related to self-esteem than were long-term donors. CONCLUSION: The 'good habit' of continued blood donation seems not to be exclusively linked to a high degree of reported other regarding ('altruistic') reasons, but also to a combination of motives, including some modestly self-regarding motives. PMID- 16262758 TI - Blood inventory management in the type and screen era. AB - BACKGROUND AND OBJECTIVE: Previous models on the management of hospital blood inventories have mostly dealt with the optimization of variables related to the assigned, cross-matched inventory. Because the type and screen (T and S) procedure allows blood banks to manage surgical reserves with only unassigned inventories, it becomes necessary to gain insight into the factors that determine the efficiency of this kind of blood inventory. STUDY DESIGN: Stochastic model that simulates the routine operation of a hospital blood bank inventory over a finite number of days was used. Factors that were analysed for their influence on outdate and shortage rates included the mean (MEAN) and variation (CVAR) in daily transfusion, the remaining shelf life of blood units shipped from the blood supplier (RSL) and the number of days between consecutive shipments (INT). RESULTS: Outdate and shortage rates grew exponentially with CVAR, an effect that could be partially counterbalanced by increasing RSL. The variables, MEAN and INT, had little influence on the inventory, provided that blood stocks shipped from the supplier are targeted at the expected average demand for transfusion and RSL is greater than INT. CONCLUSIONS: In hospitals that do not hold cross-matched inventories, CVAR is the major parameter in determining the blood inventory performance. Hospitals with large CVAR must be supplied with young red blood cell (RBC) units, whereas hospitals with smaller CVAR perform well with older stocks. These results advocate for using each hospital's CVAR as the leader parameter in regional optimization policies based on recycling blood units among participating hospitals. PMID- 16262759 TI - Molecular genetic analysis for Ax phenotype of the ABO blood group system in Chinese. AB - BACKGROUND AND OBJECTIVES: To elucidate the molecular genetic background of the Ax phenotype in the Chinese population. MATERIALS AND METHODS: The ABO genes of eight Ax phenotype samples, four Ax and four AxB, were amplified by polymerase chain reaction (PCR) and were cloned, along with those of 10 random A(1) Chinese subjects. We analysed the ABO gene transcript structure and the sequences of two exons and one intron at the ABO locus. RESULTS: Among the four Ax phenotype samples, we identified one Ax02, two Ax03 and one novel Ax allele with the 543G > T mutation in the A102 background. Two of five family members also carry the allele. Of the four AxB phenotypes, one was designated as cis-AB-1/B101; the other three were shown to carry one B allele and one O with the nt261G deletion. The B alleles of the latter three were identical to B101 except for single point mutation at nt700C > G, nt640A > G and nt641T > C, respectively. The novel B101 like alleles were first associated with A(weak)B phenotypes. CONCLUSIONS: Two ABO*B(A) alleles and an Ax allele clearly differ from all previously reported ABO alleles, suggesting that the molecular genetic background of Ax is heterogeneous in the Chinese population. PMID- 16262760 TI - Severe thrombocytopenia due to host-derived anti-HPA-1a after non-myeloablative allogeneic haematopoietic stem cell transplantation for multiple myeloma: a case report. AB - BACKGROUND AND OBJECTIVES: Host- or donor-derived alloimmune thrombocytopenia can develop after non-myeloablative allogeneic haematopoietic stem cell transplantation (HSCT). We report the first case of host-derived HPA-1a antibodies. CASE REPORT: A 52-year-old male patient received HSCT from his human leucocyte antigen (HLA)-A, -B, -C, -DR identical brother after reduced intensity conditioning. Bilinear engraftment around day 12 was accompanied by a continuous decrease of platelet counts. We investigated for platelet antibodies because of a progressive decline of platelet counts and refractoriness to platelet transfusions. METHODS: The patient's serum was tested by enzyme-linked immunosorbent assay (ELISA), a solid phase assay and monoclonal antibody-specific immobilization of platelet antigens (MAIPA) assay. Recipient's DNA from the time before HSCT and donor's DNA were genotyped for human platelet antigens. RESULTS: Serum obtained on day 15 after HSCT reacted strongly with the donor's platelets due to host-derived anti-HPA-1a- and anti-HLA I antibodies. Serum samples from days 39, 45 and 65 after HSCT contained only anti-HLA I; no antibodies were detectable on day 149. Platelet counts increased on day 20 spontaneously. The decrease of the antibodies accompanied by the increase of the platelet counts suggests progressive elimination of residual host cells. CONCLUSIONS: The HPA-1a antibodies affected thrombopoietic engraftment and the success of platelet transfusions. PMID- 16262761 TI - Light chain phenotypes of HLA antibodies in cases with suspected neonatal alloimmune thrombocytopenia. AB - BACKGROUND AND OBJECTIVES: Platelet-specific antibodies are detectable in only about 30% in suspected neonatal alloimmune thrombocytopenia (NAIT). Human leucocyte antigen (HLA) class I antibodies are often detectable, and as platelets express the corresponding antigens, it has been suggested that these antibodies can be responsible for NAIT. Light chain phenotyping may assist in the diagnosis of HLA antibodies-induced NAIT. MATERIALS AND METHODS: We determined light chain phenotypes of platelet reactive HLA antibodies in 17 sera from mothers who delivered offspring with suspected NAIT. Ten sera also contained platelet specific antibodies: HPA-1a (n = 5, one with HPA-15b), HPA-5b (n = 4) and autoantibodies (n = 1). Sera were tested for kappa or lambda restriction by monoclonal antibody-specific immobilization of platelet antigens (MAIPA). RESULTS: The cross-match with paternal platelets was positive in all cases due to HLA antibodies. We identified 5, 3 and 3 sera to contain lymphocytotoxic antibodies of single, two or multiple specificities, respectively. In six cases, HLA antibodies were only detectable by MAIPA. Light chain restriction (n = 9) was not associated with HPA containing antibodies or to any pattern of the HLA antibodies. Similarly, polyclonal antibodies (n = 8) were seen in all categories. CONCLUSION: We show that pregnancy-associated HLA antibodies can be clonal or polyclonal, irrespective of a diagnosis of NAIT. ONE-SENTENCE SUMMARY: Pregnancy associated HLA antibodies can be clonal or polyclonal, irrespective a diagnosis of NAIT. PMID- 16262762 TI - Implementation of donor screening for infectious agents transmitted by blood by nucleic acid technology in Japan. PMID- 16262763 TI - HCV and HIV-1 donor screening using nucleic acid amplification technique (NAT). PMID- 16262764 TI - Implementation of donor screening for infectious agents transmitted by blood by nucleic acid technology in Poland. PMID- 16262766 TI - Socioeconomic status modifies the sex difference in spatial skill. AB - We examined whether the male spatial advantage varies across children from different socioeconomic (SES) groups. In a longitudinal study, children were administered two spatial tasks requiring mental transformations and a syntax comprehension task in the fall and spring of second and third grades. Boys from middle- and high-SES backgrounds outperformed their female counterparts on both spatial tasks, whereas boys and girls from a low-SES group did not differ in their performance level on these tasks. As expected, no sex differences were found on the verbal comprehension task. Prior studies have generally been based on the assumption that the male spatial advantage reflects ability differences in the population as a whole. Our finding that the advantage is sensitive to variations in SES provides a challenge to this assumption, and has implications for a successful explanation of the sex-related difference in spatial skill. PMID- 16262767 TI - Affirmation of personal values buffers neuroendocrine and psychological stress responses. AB - Stress is implicated in the development and progression of a broad array of mental and physical health disorders. Theory and research on the self suggest that self-affirming activities may buffer these adverse effects. This study experimentally investigated whether affirmations of personal values attenuate physiological and psychological stress responses. Eighty-five participants completed either a value-affirmation task or a control task prior to participating in a laboratory stress challenge. Participants who affirmed their values had significantly lower cortisol responses to stress, compared with control participants. Dispositional self-resources (e.g., trait self-esteem and optimism) moderated the relation between value affirmation and psychological stress responses, such that participants who had high self-resources and had affirmed personal values reported the least stress. These findings suggest that reflecting on personal values can keep neuroendocrine and psychological responses to stress at low levels. Implications for research on the self, stress processes, health, and interventions are discussed. PMID- 16262768 TI - The interaction of payoff structure and regulatory focus in classification. AB - This report brings together research on motivation and learning by exploring how fit of regulatory focus affects people's ability to acquire new categories. Perceptual categories were learned by people with a promotion focus (a situationally determined sensitivity to gains) or a prevention focus (a sensitivity to losses). Classification performance was closest to optimal (as determined by models fit to individual subjects' data) when the regulatory focus matched the structure of the payoffs for the categories. Promotion-focus subjects performed best when the payoffs consisted of all gains. Prevention-focus subjects performed best when the payoffs consisted of all losses. PMID- 16262769 TI - "Tip of the fingers" experiences by deaf signers: insights into the organization of a sign-based lexicon. AB - The "tip of the fingers" phenomenon (TOF) for sign language parallels the "tip of the tongue" phenomenon (TOT) for spoken language. During a TOF, signers are sure they know a sign but cannot retrieve it. Although some theories collapse semantics and phonology in sign language and thus predict that TOFs should not occur, TOFs were elicited in the current study. Like TOTs, TOFs often resolve spontaneously, commonly involve targets that are proper names, and frequently include partial access to phonology. Specifically, signers were more likely to retrieve a target sign's handshape, location, and orientation than to retrieve its movement. Signers also frequently recalled the first letter of a finger spelled word. The existence of TOFs supports two-stage retrieval and a division between semantics and phonology in American Sign Language. The partial phonological information available during TOFs suggests that phonological features are accessed more simultaneously during lexical access for signed language than during lexical access for spoken language. PMID- 16262770 TI - Predicting literacy at age 7 from preliteracy at age 4. AB - Early literacy experience and preliteracy knowledge have been shown to predict later literacy outcomes. Using a representative sample of 3,052 same-sex twin pairs (6,104 children) in the United Kingdom, we explored phenotypic and etiological interrelationships among early literacy experience, preliteracy knowledge, and school-based literacy outcomes (reading and writing). Both literacy experience and preliteracy knowledge at age 4 significantly and independently predicted literacy at age 7. Both measures also showed genetic influence that significantly predicted literacy at age 7, although genetic mediation was stronger for preliteracy knowledge than for early literacy experience. However, for both measures, shared environmental factors explained most of the association with literacy at age 7. PMID- 16262771 TI - Is developing scientific thinking all about learning to control variables? AB - Academically low-performing urban sixth graders engaged in inquiry activity received a suggestion that they focus their investigation on the role of a single factor. This suggestion had significant effects on their use of a superficially dissimilar strategy--controlling the variation of other factors. This latter strategy has received the lion's share of attention in research on the development of scientific reasoning. These results have implications, we propose, for what undergoes development with respect to scientific thinking and how this development can best be facilitated. PMID- 16262772 TI - Early science instruction: addressing fundamental issues. PMID- 16262773 TI - What needs to be mastered in mastery of scientific method? PMID- 16262774 TI - "We all look the same to me": positive emotions eliminate the own-race in face recognition. AB - Extrapolating from the broaden-and-build theory, we hypothesized that positive emotion may reduce the own-race bias in facial recognition. In Experiments 1 and 2, Caucasian participants (N = 89) viewed Black and White faces for a recognition task. They viewed videos eliciting joy, fear, or neutrality before the learning (Experiment 1) or testing (Experiment 2) stages of the task. Results reliably supported the hypothesis. Relative to fear or a neutral state, joy experienced before either stage improved recognition of Black faces and significantly reduced the own-race bias. Discussion centers on possible mechanisms for this reduction of the own-race bias, including improvements in holistic processing and promotion of a common in-group identity due to positive emotions. PMID- 16262775 TI - The effects of reward and punishment in violent video games on aggressive affect, cognition, and behavior. AB - Three experiments examined the effects of rewarding and punishing violent actions in video games on later aggression-related variables. Participants played one of three versions of the same race-car video game: (a) a version in which all violence was rewarded, (b) a version in which all violence was punished, and (c) a nonviolent version. Participants were then measured for aggressive affect (Experiment 1), aggressive cognition (Experiment 2), and aggressive behavior (Experiment 3). Rewarding violent game actions increased hostile emotion, aggressive thinking, and aggressive behavior. Punishing violent actions increased hostile emotion, but did not increase aggressive thinking or aggressive behavior. Results suggest that games that reward violent actions can increase aggressive behavior by increasing aggressive thinking. PMID- 16262776 TI - Perceiving sex directly and indirectly: meaning in motion and morphology. AB - We employed a novel technique to explore how the body's motion and morphology affect judgments of sex and gender. Stimuli depicted animated human walkers that varied in motion (gait patterns varying shoulder swagger and hip sway) and in morphology (waist-to-hip ratio). The potency of morphology in categorical sex judgments was confirmed. Visual scanning of the walkers was concentrated in the waist and hip region of the body (Study 1a). This targeted scanning was attenuated, however, when the sex of the target had been prespecified (Study 1b). Body motion permitted categorical judgments of sex, but these judgments were mediated by perceived gender (Study 2). These studies provide converging evidence for the primacy of the body's shape in categorical judgments of sex. PMID- 16262777 TI - Shadows of the past: temporal retrieval effects in recognition memory. AB - We examine whether temporally defined associations play a role in item recognition. The role of these associations in recall tasks is well known; we demonstrate an important role in item recognition as well. In this study, subjects were significantly more likely to recognize a test item as having been previously experienced if the preceding test item was studied in a temporally proximal list position than if the preceding test item came from a more distant list position. Further analyses showed that this associative effect was almost entirely due to cases in which the preceding test item received a highest confidence recognition judgment. PMID- 16262778 TI - Multiple object tracking in people with Williams syndrome and in normally developing children. AB - Multiple object tracking is hypothesized to utilize visual indexes, which may provide rapid, parallel access to a limited number of visual objects, thereby supporting a variety of spatial tasks. We examined whether faulty indexing might play a role in the severe visuospatial deficits found in Williams syndrome. We asked observers to track from one to four targets in a display of eight identical objects. Objects remained stationary (static condition) or moved randomly and independently (moving condition) for 6 s, after which observers pointed to the objects they thought were targets. People with Williams syndrome were impaired in the moving condition, but not the static condition, compared with mental-age matched control participants. Normal children who were younger than the mental age-matched control children did not show the same profile as individuals with Williams syndrome, which suggests that the difference between the tasks in Williams syndrome did not reflect simple developmental immaturity. Error analysis revealed that all groups had "slippery" indexes, falsely identifying target neighbors, and further suggested that people with Williams syndrome deploy fewer indexes than do people without this disorder. PMID- 16262779 TI - What children are looking at during shared storybook reading. AB - Two studies were conducted to determine the extent to which young children fixate on the print of storybooks during shared book reading. Children's books varying in the layout of the print and the richness of the illustrations were displayed on a computer monitor. Each child's mother or preschool teacher read the books while the child sat on the adult's lap wearing an EyeLink headband that recorded visual fixations. In both studies, children spent very little time examining the print regardless of the nature of the print and illustrations. Although fixations on the illustrations were highly correlated with the length of the accompanying text and could be altered by altering the content of the text, fixations to the text were uncorrelated with the length of the text. These results indicate that preschool children engage in minimal exploration of the print during shared book reading. PMID- 16262780 TI - The MreB and Min cytoskeletal-like systems play independent roles in prokaryotic polar differentiation. AB - Establishment of an axis of cell polarity and differentiation of the cell poles are fundamental aspects of cellular development in many organisms. We compared the effects of two bacterial cytoskeletal-like systems, the MreB and MinCDE systems, on these processes in Escherichia coli. We report that the Min proteins are capable of establishing an axis of oscillation that is the initial step in establishment of polarity in spherical cells, in a process that is independent of the MreB cytoskeleton. In contrast, the MreB system is required for establishment of the rod shape of the cell and for polar targeting of other polar constituents, such as the Shigella virulence factor IcsA and the aspartate chemoreceptor Tar, in a process that is independent of the Min system. Thus, the two bacterial cytoskeletal-like systems act independently on different aspects of cell polarization. PMID- 16262781 TI - SsgA-like proteins determine the fate of peptidoglycan during sporulation of Streptomyces coelicolor. AB - During developmental cell division in sporulation-committed aerial hyphae of streptomycetes, up to a hundred septa are simultaneously produced, in close harmony with synchromous chromosome condensation and segregation. Several unique protein families are involved in the control of this process in actinomycetes, including that of the SsgA-like proteins (SALPs). Mutants for each of the individual SALP genes were obtained, and high-resolution and fluorescence imaging revealed that each plays an important and highly specific role in the control of the sporulation process, and their function relates to the build-up and degradation of septal and spore-wall peptidoglycan. While SsgA and SsgB are essential for sporulation-specific cell division in Streptomyces coelicolor, SsgC G are responsible for correct DNA segregation/condensation (SsgC), spore wall synthesis (SsgD), autolytic spore separation (SsgE, SsgF) or exact septum localization (SsgG). Our experiments paint a picture of a novel protein family that acts through timing and localization of the activity of penicillin-binding proteins and autolysins, thus controlling important steps during the initiation and the completion of sporulation in actinomycetes. PMID- 16262782 TI - Efficient secretion of a folded protein domain by a monomeric bacterial autotransporter. AB - Bacterial autotransporters are proteins that contain a small C-terminal 'beta domain' that facilitates translocation of a large N-terminal 'passenger domain' across the outer membrane (OM) by an unknown mechanism. Here we used EspP, an autotransporter produced by Escherichia coli 0157:H7, as a model protein to gain insight into the transport reaction. Initially we found that the passenger domain of a truncated version of EspP (EspPDelta1-851) was translocated efficiently across the OM. Blue Native polyacrylamide gel electrophoresis, analytical ultracentrifugation and other biochemical methods showed that EspPDelta1-851 behaves as a compact monomer and strongly suggest that the channel formed by the beta domain is too narrow to accommodate folded polypeptides. Surprisingly, we found that a folded protein domain fused to the N-terminus of EspPDelta1-851 was efficiently translocated across the OM. Further analysis revealed that the passenger domain of wild-type EspP also folds at least partially in the periplasm. To reconcile these data, we propose that the EspP beta domain functions primarily to target and anchor the protein and that an external factor transports the passenger domain across the OM. PMID- 16262783 TI - The ExPortal: an organelle dedicated to the biogenesis of secreted proteins in Streptococcus pyogenes. AB - The Gram-positive pathogen Streptococcus pyogenes secretes proteins through the ExPortal, a unique single microdomain of the cellular membrane specialized to contain the Sec translocons. It has been proposed that the ExPortal functions as an organelle to promote the biogenesis of secreted proteins by coordinating interactions between nascent unfolded secretory proteins and membrane-associated chaperones. In this study we provide evidence to support this model. It was found that HtrA (DegP), a surface anchored accessory factor required for maturation of the secreted SpeB cysteine protease, was localized exclusively to the ExPortal. Furthermore, the ATP synthase beta subunit was not localized to the ExPortal, suggesting that retention is likely restricted to a specific subset of exported proteins. Mutations that disrupted the anchoring, but not the protease activity, of HtrA, also altered the maturation kinetics of SpeB demonstrating that localization to the ExPortal was important for HtrA function. These data indicate that the ExPortal provides a mechanism by which Gram-positive bacteria can coordinate protein secretion and subsequent biogenesis in the absence of a specialized protein-folding compartment. PMID- 16262784 TI - The flagellar hierarchy of Rhodobacter sphaeroides is controlled by the concerted action of two enhancer-binding proteins. AB - The expression of the bacterial flagellar genes follows a hierarchical pattern. In Rhodobacter sphaeroides the flagellar genes encoding the hook and basal body proteins are expressed from sigma54-dependent promoters. This type of promoters is always regulated by transcriptional activators that belong to the family of the enhancer-binding proteins (EBPs). We searched for possible EBPs in the genome of R. sphaeroides and mutagenized two open reading frames (ORFs) (fleQ and fleT), which are in the vicinity of flagellar genes. The resulting mutants were non motile and could only be complemented by the wild-type copy of the mutagenized gene. Transcriptional fusions showed that all the flagellar sigma54-dependent promoters with exception of fleTp, required both transcriptional activators for their expression. Interestingly, transcription of the fleT operon is only dependent on FleQ, and FleT has a negative effect. Both activators were capable of hydrolysing ATP, and were capable of promoting transcription from the flagellar promoters at some extent. Electrophoretic mobility shift assays suggest that only FleQ interacts with DNA whereas FleT improves binding of FleQ to DNA. A four-tiered flagellar transcriptional hierarchy and a regulatory mechanism based on the intracellular concentration of both activators and differential enhancer affinities are proposed. PMID- 16262786 TI - A novel palmitoyl acyl transferase controls surface protein palmitoylation and cytotoxicity in Giardia lamblia. AB - The intestinal protozoan parasite Giardia lamblia undergoes surface antigenic variation whereby one of a family of structurally related variant-specific surface proteins (VSPs) is replaced in a regulated process by another antigenically distinct VSP. All VSPs are type I membrane proteins that have a conserved hydrophobic sequence terminated by the invariant hydrophilic amino acids, CRGKA. Using transfected Giardia constitutively expressing HA-tagged VSPH7 and incubated with radioactive [3H]palmitate, we demonstrate that the palmitate is attached to the Cys in the conserved CRGKA tail. Surface location of mutant VSPs lacking either the CRGKA tail or its Cys is identical to that of wild-type VSPH7 but non-palmitoylated mutants fail to undergo complement-independent antibody specific cytotoxicity. In addition, membrane localization of non palmitoylated mutant VSPH7 changes from a pattern similar to rafts to non-rafts. Palmitoyl transferases (PAT), responsible for protein palmitoylation in other organisms, often possess a cysteine-rich domain containing a conserved DHHC motif (DHHC-CRD). An open reading frame corresponding to a putative 50 kDa Giardia PAT (gPAT) containing a DHHC-CRD motif was found in the Giardia genome database. Expression of epitope-tagged gPAT using a tetracycline inducible vector localized gPAT to the plasma membrane, a pattern similar to that of VSPs. Transfection with gPAT antisense producing vectors inhibits gPAT expression and palmitoylation of VSPs in vitro confirming the function of gPAT. These results show that VSPs are palmitoylated at the cysteine within the conserved tail by gPAT and indicate an essential function of palmitoylation in control of VSP-mediated signalling and processing. PMID- 16262785 TI - Candida albicans serotype B strains synthesize a serotype-specific phospholipomannan overexpressing a beta-1,2-linked mannotriose. AB - Candida albicans strains consist of serotypes A and B depending on the presence of terminal beta-1,2-linked mannose residues in the acid-stable part of serotype A phosphopeptidomannan (PPM). The distribution of C. albicans serotypes varies according to country and human host genetic and infectious backgrounds. However, these epidemiological traits have not yet been related to a phenotypically stable molecule as cell surface expression of the serotype A epitope depends on the growth conditions. We have shown that C. albicans serotype A associates beta mannose residues with another molecule, phospholipomannan (PLM), which is a member of the mannoseinositolphosphoceramide family. In this study, PLM from serotype B strains was analysed in order to provide structural bases for the differences in molecular mass and antigenicity observed between PLMs from both serotypes. Through these analyses, carbon 10 was shown to be the location of a second hydroxylation of fatty acids previously unknown in fungal sphingolipids. Minor differences observed in the ceramide moiety appeared to be strain dependent. More constant features of PLM from serotype B strains were the incorporation of greater amounts of phytosphingosine C20, a twofold reduced glycosylation of PLM and overexpression of a beta-1,2 mannotriose, the epitope of protective antibodies. This specific beta-mannosylation was observed even when growth conditions altered serotype A PPM-specific epitopes, confirming the potential of PLM as a phenotypically stable molecule for serotyping. This study also suggests that the regulation of beta-mannosyltransferases, which define specific immunomodulatory adhesins whose activity depends on the mannosyl chain length, are part of the genetic background that differentiates serotypes. PMID- 16262787 TI - Common beta-lactamases inhibit bacterial biofilm formation. AB - Beta-lactamases, which evolved from bacterial penicillin-binding proteins (PBPs) involved in peptidoglycan (PG) synthesis, confer resistance to beta-lactam antibiotics. While investigating the genetic basis of biofilm development by Pseudomonas aeruginosa, we noted that plasmid vectors encoding the common beta lactamase marker TEM-1 caused defects in twitching motility (mediated by type IV pili), adherence and biofilm formation without affecting growth rates. Similarly, strains of Escherichia coli carrying TEM-1-encoding vectors grew normally but showed reduced adherence and biofilm formation, showing this effect was not species-specific. Introduction of otherwise identical plasmid vectors carrying tetracycline or gentamicin resistance markers had no effect on biofilm formation or twitching motility. The effect is restricted to class A and D enzymes, because expression of the class D Oxa-3 beta-lactamase, but not class B or C beta lactamases, impaired biofilm formation by E. coli and P. aeruginosa. Site directed mutagenesis of the catalytic Ser of TEM-1, but not Oxa-3, abolished the biofilm defect, while disruption of either TEM-1 or Oxa-3 expression restored wild-type levels of biofilm formation. We hypothesized that the A and D classes of beta-lactamases, which are related to low molecular weight (LMW) PBPs, may sequester or alter the PG substrates of such enzymes and interfere with normal cell wall turnover. In support of this hypothesis, deletion of the E. coli LMW PBPs 4, 5 and 7 or combinations thereof, resulted in cumulative defects in biofilm formation, similar to those seen in beta-lactamase-expressing transformants. Our results imply that horizontal acquisition of beta-lactamase resistance enzymes can have a phenotypic cost to bacteria by reducing their ability to form biofilms. Beta-lactamases likely affect PG remodelling, manifesting as perturbation of structures involved in bacterial adhesion that are required to initiate biofilm formation. PMID- 16262788 TI - AIP56, a novel plasmid-encoded virulence factor of Photobacterium damselae subsp. piscicida with apoptogenic activity against sea bass macrophages and neutrophils. AB - A strategy used by extracellular pathogens to evade phagocytosis is the utilization of exotoxins that kill host phagocytes. We have recently shown that one major pathogenicity strategy of Photobacterium damselae subsp. piscicida (Phdp), the agent of the widespread fish pasteurellosis, is the induction of extensive apoptosis of sea bass macrophages and neutrophils that results in lysis of these phagocytes by post-apoptotic secondary necrosis. Here we show that this unique process is mediated by a novel plasmid-encoded apoptosis inducing protein of 56 kDa (AIP56), an exotoxin abundantly secreted by all virulent, but not avirulent, Phdp strains tested. AIP56 is related to an unknown protein of the enterohemorrhagic Escherichia coli O157:H7 and NleC, a Citrobacter rodentium type III secreted effector of unknown function. Passive immunization of sea bass with a rabbit anti-AIP56 serum conferred protection against Phdp challenge, indicating that AIP56 represents a key virulence factor of that pathogen and is a candidate for the design of an anti-pasteurellosis vaccine. PMID- 16262789 TI - Trafficking determinants for PfEMP3 export and assembly under the Plasmodium falciparum-infected red blood cell membrane. AB - During the maturation of intracellular asexual stages of Plasmodium falciparum parasite-encoded proteins are exported into the erythrocyte cytosol. A number of these parasite proteins attach to the host cell cytoskeleton and facilitate transformation of a disk-shaped erythrocyte into a rounded and more rigid infected erythrocyte able to cytoadhere to the vasculature. Knob formation on the surface of infected erythrocytes is critical for this cytoadherence to the host endothelium. P. falciparum proteins have been identified that localize to the parasite-infected erythrocyte membrane: the variant cytoadherence ligand erythrocyte membrane protein 1 (PfEMP1), the knob-associated histidine-rich protein (KAHRP) and the erythrocyte membrane protein 3 (PfEMP3). In this study, we have generated parasites expressing PfEMP3-green fluorescent protein chimeras and identified domains involved in entry to the secretory pathway, export across the parasitophorous vacuolar membrane and attachment to Maurer's clefts and the erythrocyte membrane. Solubility assays, fluorescence photobleaching experiments and immunogold electron microscopy suggest that the exported chimeric proteins are trafficked in a complex rather than in vesicles. This study characterizes elements involved in the tight but transient binding of PfEMP3 to Maurer's clefts and shows that the same elements are necessary for correct assembly under the erythrocyte membrane. PMID- 16262790 TI - Identification of Klebsiella pneumoniae virulence determinants using an intranasal infection model. AB - Klebsiella pneumoniae is a Gram-negative enterobacterium that has historically been, and currently remains, a significant cause of human disease. It is a frequent cause of urinary tract infections and pneumonia, and subsequent systemic infections can have mortality rates as high as 60%. Despite its clinical significance, few virulence factors of K. pneumoniae have been identified or characterized. In this study we present a mouse model of acute K. pneumoniae respiratory infection using an intranasal inoculation method, and examine the progression of both pulmonary and systemic disease. Wild-type infection recapitulates many aspects of clinical disease, including significant bacterial growth in both the trachea and lungs, an inflammatory immune response characterized by dramatic neutrophil influx, and a steady progression to systemic disease with ensuing mortality. These observations are contrasted with an infection by an isogenic capsule-deficient strain that shows an inability to cause disease in either pulmonary or systemic tissues. The consistency and clinical accuracy of the intranasal mouse model proved to be a useful tool as we conducted a genetic screen to identify novel virulence factors of K. pneumoniae. A total of 4800 independent insertional mutants were evaluated using a signature tagged mutagenesis protocol. A total of 106 independent mutants failed to be recovered from either the lungs or spleens of infected mice. Small scale independent infections proved to be helpful as a secondary screening method, as opposed to the more traditional competitive index assay. Those mutants showing verified attenuation contained insertions in loci with a variety of putative functions, including a large number of hypothetical open reading frames. Subsequent experiments support the premise that the central mechanism of K. pneumoniae pathogenesis is the production of a polysaccharide-rich cell surface that provides protection from the inflammatory response. PMID- 16262792 TI - Identification and molecular characterization of an N-acetylmuramyl-L-alanine amidase Sle1 involved in cell separation of Staphylococcus aureus. AB - We purified a peptidoglycan hydrolase involved in cell separation from a Staphylococcus aureus atl null mutant and identified its gene. Characterization of the gene product shows a 32 kDa N-acetylmuramyl-L-alanine amidase that we designated Sle1. Analysis of peptidoglycan digests showed Sle1 preferentially cleaved N-acetylmuramyl-L-Ala bonds in dimeric cross-bridges that interlink the two murein strands in the peptidoglycan. An insertion mutation of sle1 impaired cell separation and induced S. aureus to form clusters suggesting Sle1 is involved in cell separation of S. aureus. The Sle1 mutant revealed a significant decrease in pathogenesis using an acute infection mouse model. Atl is the major autolysin of S. aureus, which has been implicated in cell separation of S. aureus. Generation of an atl/sle1 double mutant revealed that the mutant cell separation was heavily impaired suggesting that S. aureus uses two peptidoglycan hydrolases, Atl and Sle1, for cell separation. Unlike Atl, Sle1 is not directly involved in autolysis of S. aureus. PMID- 16262791 TI - Identification of novel single amino acid changes that result in hyperactivation of the unique GTPase, Rheb, in fission yeast. AB - Rheb GTPase is a key player in the control of growth, cell cycle and nutrient uptake that is conserved from yeast to humans. To further our understanding of the Rheb pathway, we sought to identify hyperactivating mutations in the Schizosaccharomyces pombe Rheb, Rhb1. Hyperactive forms of Rhb1 were found to result from single amino acid changes at valine-17, serine-21, lysine-120 or asparagine-153. Expression of these mutants confers resistance to canavanine and thialysine, phenotypes which are similar to phenotypes exhibited by cells lacking the Tsc1/Tsc2 complex that negatively regulates Rhb1. The thialysine-resistant phenotype of the hyperactive Rhb1 mutants is suppressed by a second mutation in the effector domain. Purified mutant proteins exhibit dramatically decreased binding of GDP, while their GTP binding is not drastically affected. In addition, some of the mutant proteins show significantly decreased GTPase activities. Thus the hyperactivating mutations are expected to result in an increase in the GTP bound/GDP-bound ratio of Rhb1. By using the hyperactive mutant, Rhb1(K120R), we have been able to demonstrate that Rhb1 interacts with Tor2, one of the two S. pombe TOR (Target of Rapamycin) proteins. These fission yeast results provide the first evidence for a GTP-dependent association of Rheb with Tor. PMID- 16262793 TI - Two different polyketide synthase genes are required for synthesis of zearalenone in Gibberella zeae. AB - Zearalenone (ZEA) is a polyketide mycotoxin produced by some species of Gibberella/Fusarium and causes hyperestrogenic syndrome in animals. ZEA occurs naturally in cereals infected by Gibberella zeae in temperate regions and threatens animal health. In this study, we report on a set of genes that participate in the biosynthesis of ZEA in G. zeae. Focusing on the non-reducing polyketide synthase (PKS) genes of the G. zeae genome, we demonstrated that PKS13 is required for ZEA production. Subsequent analyses revealed that a continuous, 50 kb segment of DNA carrying PKS13 consisted of three additional open reading frames that were coexpressed as a cluster during the condition for ZEA biosynthesis. These genes, in addition to PKS13, were essential for the ZEA biosynthesis. They include another PKS gene (PKS4) encoding a fungal reducing PKS; zearalenone biosynthesis gene 1 (ZEB1), which shows a high similarity to putative isoamyl alcohol oxidase genes; and ZEB2 whose deduced product carries a conserved, basic-region leucine zipper domain. ZEB1 is responsible for the chemical conversion of beta-zearalenonol (beta-ZOL) to ZEA in the biosynthetic pathway, and ZEB2 controls transcription of the cluster members. Transcription of these genes was strongly influenced by different culture conditions such as nutrient starvations and ambient pH. Furthermore, the same set of genes regulated by ZEB2 was dramatically repressed in the transgenic G. zeae strain with the deletion of PKS13 or PKS4 but not in the ZEB1 deletion strain, suggesting that ZEA or beta-ZOL may be involved in transcriptional activation of the gene cluster required for ZEA biosynthesis in G. zeae. This is the first published report on the molecular characterization of genes required for ZEA biosynthesis. PMID- 16262794 TI - RepB protein of an Agrobacterium tumefaciens Ti plasmid binds to two adjacent sites between repA and repB for plasmid partitioning and autorepression. AB - Plasmids of Agrobacterium tumefaciens replicate using the products of the repABC operon, which are highly conserved among plasmids and some chromosomes of the alpha-Proteobacteria. The products of repA and repB direct plasmid partitioning, while the repC gene encodes a replication initiator protein. The transcription of the repABC operon of tumour inducing (Ti) plasmids is both negatively autoregulated by the RepA and RepB proteins, and positively regulated by TraR. In the present study, we have identified a fourth gene (repD) in the repABC operon of an octopine-type Ti plasmid. repD is 78 codons in length, and maps between repA and repB genes. A repD-lacZ protein fusion demonstrated that repD is strongly expressed. Two identical binding sites for the RepB protein were found within the repD coding sequence, and these sites are required for plasmid stability and for maximal repression of repABC transcription. RepA protein enhances the binding of RepB at these binding sites, just as RepB increases the affinity of RepA for binding sites at the repABC P4 promoter. We propose that RepA and RepB form complexes that bind both sites, possibly causing a loop that is important for repression of the repABC operon. Binding at one or both sites may also be required for accurate plasmid partitioning. PMID- 16262795 TI - The VirE1VirE2 complex of Agrobacterium tumefaciens interacts with single stranded DNA and forms channels. AB - The VirE2 protein is crucial for the transfer of single-stranded DNA (ssDNA) from Agrobacterium tumefaciens to the nucleus of the plant host cell because of its ssDNA binding activity, assistance in nuclear import and putative ssDNA channel activity. The native form of VirE2 in Agrobacterium's cytoplasm is in complex with its specific chaperone, VirE1. Here, we describe the ability of the VirE1VirE2 complex to both bind ssDNA and form channels. The affinity of the VirE1VirE2 complex for ssDNA is slightly reduced compared with VirE2, but the kinetics of binding to ssDNA are unaffected by the presence of VirE1. Upon binding of VirE1VirE2 to ssDNA, similar helical structures to those reported for the VirE2-ssDNA complex were observed by electron microscopy. The VirE1VirE2 complex can release VirE1 once the VirE2-ssDNA complexes assembled. VirE2 exhibits a low affinity for small unilamellar vesicles composed of bacterial lipids and a high affinity for lipid vesicles containing sterols and sphingolipids, typical components of animal and plant membranes. In contrast, the VirE1VirE2 complex associated similarly with all kind of lipids. Finally, black lipid membrane experiments revealed the ability of the VirE1VirE2 complex to form channels. However, the majority of the channels displayed a conductance that was a third of the conductance of VirE2 channels. Our results demonstrate that the binding of VirE1 to VirE2 does not inhibit VirE2 functions and that the effector chaperone complex is multifunctional. PMID- 16262796 TI - Characterization of functional domains of the Vibrio cholerae virulence regulator ToxT. AB - The toxT gene encodes an AraC family transcriptional activator that is responsible for regulating virulence gene expression in Vibrio cholerae. Analysis of ToxT by dominant/negative assays and a LexA-based reporter system demonstrated that the N-terminus of the protein contains dimerization determinants, indicating that ToxT likely functions as a dimer. Additionally, a natural variant of ToxT with only 60% identity in the N-terminus, as well as a mutant form of ToxT with an altered amino acid in the N-terminus (L107F), exhibited altered transcriptional responses to bile, suggesting that the N-terminus is involved in environmental sensing. The C-terminus of ToxT functions to bind DNA and requires dimerization for stable binding in vitro, as demonstrated by gel shift analysis. Interestingly, a dimerized form of the ToxT C-terminus is able to bind DNA in vitro but is transcriptionally inactive in vivo, indicating that the N-terminus contains determinants that are required for transcriptional activation. These results provide a model for a two-domain structure for ToxT, with an N-terminal dimerization and environmental sensing domain and a C-terminal DNA-binding domain; unlike other well-studied AraC family proteins, both domains of ToxT appear to be required for transcriptional activation. PMID- 16262797 TI - All four Mycobacterium tuberculosis glnA genes encode glutamine synthetase activities but only GlnA1 is abundantly expressed and essential for bacterial homeostasis. AB - Glutamine synthetases (GS) are ubiquitous enzymes that play a central role in every cell's nitrogen metabolism. We have investigated the expression and activity of all four genomic Mycobacterium tuberculosis GS - GlnA1, GlnA2, GlnA3 and GlnA4 - and four enzymes regulating GS activity and/or nitrogen and glutamate metabolism - adenylyl transferase (GlnE), gamma-glutamylcysteine synthase (GshA), UDP-N-acetylmuramoylalanine-D-glutamate ligase (MurD) and glutamate racemase (MurI). All eight genes are located in multigene operons except for glnA1, and all are transcribed in M. tuberculosis; however, some are not translated or translated at such low levels that the enzymes escape detection. Of the four GS, only GlnA1 can be detected. Each of the eight genes, as well as the glnA1-glnE glnA2 cluster, was expressed separately in Mycobacterium smegmatis, and its gene product was characterized and assayed for enzymatic activity by analysing the reaction products. In M. smegmatis, all four recombinant-overexpressed GS are multimeric enzymes exhibiting GS activity. Whereas GlnA1, GlnA3 and GlnA4 catalyse the synthesis of L-glutamine, GlnA2 catalyses the synthesis of D glutamine and D-isoglutamine. The generation of mutants in M. tuberculosis of the four glnA genes, murD and murI demonstrated that all of these genes except glnA1 are nonessential for in vitro growth. L-methionine-S,R-sulphoximine (MSO), previously demonstrated to inhibit M. tuberculosis growth in vitro and in vivo, strongly inhibited all four GS enzymes; hence, the design of MSO analogues with an improved therapeutic to toxic ratio remains a promising strategy for the development of novel anti-M. tuberculosis drugs. PMID- 16262798 TI - Sialic acid transport in Haemophilus influenzae is essential for lipopolysaccharide sialylation and serum resistance and is dependent on a novel tripartite ATP-independent periplasmic transporter. AB - Sialylation of the lipopolysaccharide (LPS) is an important mechanism used by the human pathogen Haemophilus influenzae to evade the innate immune response of the host. We have demonstrated that N-acetylneuraminic acid (Neu5Ac or sialic acid) uptake in H. influenzae is essential for the subsequent modification of the LPS and that this uptake is mediated through a single transport system which is a member of the tripartite ATP-independent periplasmic (TRAP) transporter family. Disruption of either the siaP (HI0146) or siaQM (HI0147) genes, that encode the two subunits of this transporter, results in a complete loss of uptake of [14C] Neu5Ac. Mutant strains lack sialylated glycoforms in their LPS and are more sensitive to killing by human serum than the parent strain. The SiaP protein has been purified and demonstrated to bind a stoichiometric amount of Neu5Ac by electrospray mass spectrometry. This binding was of high affinity with a Kd of approximately 0.1 microM as determined by protein fluorescence. The inactivation of the SiaPQM TRAP transporter also results in decreased growth of H. influenzae in a chemically defined medium containing Neu5Ac, supporting an additional nutritional role of sialic acid in H. influenzae physiology. PMID- 16262801 TI - Coming to a medical school near you: full motion video medical education. PMID- 16262799 TI - CsrA and three redundant small RNAs regulate quorum sensing in Vibrio cholerae. AB - Bacteria communicate using a process called quorum sensing which involves production, secretion and detection of signalling molecules called autoinducers. Quorum sensing allows populations of bacteria to simultaneously regulate gene expression in response to changes in cell density. The human pathogen, Vibrio cholerae, uses a quorum-sensing circuit composed of parallel systems that transduce information through four redundant regulatory small RNAs (sRNAs) called quorum regulatory RNAs (Qrr) to control the expression of numerous genes, most notably those required for virulence. We show that the VarS/VarA two-component sensory system comprises an additional regulatory input controlling quorum sensing-dependent gene expression in V. cholerae. VarS/VarA controls transcription of three previously unidentified small regulatory RNAs (sRNAs) that are similar to the sRNAs CsrB and CsrC of Escherichia coli. The three V. cholerae sRNAs, which we name CsrB, CsrC and CsrD, act redundantly to control the activity of the global regulatory protein, CsrA. The VarS/VarA-CsrA/BCD system converges with the V. cholerae quorum-sensing systems to regulate the expression of the Qrr sRNAs, and thus, the entire quorum-sensing regulon. PMID- 16262802 TI - The significance of de-roling and debriefing in training medical students using simulation to train medical students. PMID- 16262804 TI - Assessment of the quality of interaction in distance learning programmes utilising the Internet (WebCT) or interactive television (ITV). AB - INTRODUCTION: This study focuses on the quality of interaction in interactive TV (ITV), WebCT bulletin boards (BBs) and chat rooms (CRs) and addresses the question of how effectively new collaborative electronic technologies have been married with new pedagogical ideas to create effective learning for distance education students. METHODS: Fifteen (out of 68) BB, 14 (out of 32) CR and 13 (out of 25) ITV conversations were randomly selected for coding using a modified exchange structure analysis. The roles that students and lecturers took in the conversations were determined from this. RESULTS: The percentage of turns made by lecturers as opposed to students was 51% in CRs, 14% in BBs and 68% in ITV. The percentage of turns spent on actual coursework was 73% in CRs, 89% in BBs and 82% in ITV. Comparisons between tutors' and students' roles within as well as between ITV, BBs and CRs were all statistically significant with P < 0.05. In CRs the main roles of both students and lecturers were those of elaborators, inquirers and explainers. In BBs the main roles of students and lecturers were those of explainers and evaluators. In ITV sessions students' main roles were those of elaborators and explainers, whereas lecturers' main roles were those of lecturers, elaborators, inquirers and evaluators. CONCLUSION: In terms of creating a constructivist and active learning community that can operate within a distance learning paradigm, WebCT appears superior to ITV. PMID- 16262803 TI - Comparison of text and video cases in a postgraduate problem-based learning format. AB - OBJECTIVE: To investigate whether adding a brief video case instead of an equivalent written text improves the cognitive and metacognitive processes (data exploration, theory building, theory evaluation and metareasoning) of residents in problem-based learning. SUBJECTS: Paediatric residents. SETTING: Department of Paediatrics, Arhus University Hospital, Skejby, Denmark. METHODS: Eleven residents were each assigned to 1 of 2 groups. Both groups analysed an identical vignette of a patient case. Immediately after this, 1 group watched a 2.5-minute video recording and the other group read a description of the same video recording. The groups then reanalysed the case. Thinking processes were tapped by recording and analysing the verbal group interaction. The frequencies of clause categories were calculated and compared using chi-square tests. RESULTS: The verbal interaction showed statistically significant improvements in data exploration, theory building and theory evaluation after the video case. CONCLUSIONS: The greater improvement in clause frequency after the video case compared with the equivalent text suggests an improvement in data exploration, theory building and theory evaluation. The use of patient video recordings may be a valuable supplement to postgraduate paediatric training, particularly pertaining to movement disorders. PMID- 16262805 TI - Survey on Aboriginal issues within Canadian medical programmes. AB - INTRODUCTION: Medical programmes across Canada recognise the increased need for Aboriginal doctors. This study aimed to determine Canadian medical programmes' commitment towards Aboriginal health issues, recruitment, admission policies, educational opportunities and support offered to Aboriginal medical undergraduate students. METHODS: Medical school websites were initially reviewed to identify recruitment initiatives, admission policies and curriculum activities related to Aboriginal health. A questionnaire was sent to each dean of medicine to collect additional data on the programmes' recruitment strategies, admission policies, educational opportunities and the type of support offered to Aboriginal medical students. RESULTS: Sixteen medical programmes completed the questionnaire (return rate = 100%). There were 56 Aboriginal medical students enrolled across these medical programmes. More than 75% of students were completing their training in a western province. Over half of the medical programmes had recruitment initiatives and admission policies specific to Aboriginal applicants and the majority reported that their programmes' curricula included learning objectives specific to Aboriginal health. Most offered lectures and training opportunities to medical students and few offered core curriculum activities focusing on Aboriginal health. DISCUSSION: This descriptive paper offers a snapshot of initiatives across medical programmes aimed at increasing the number of Aboriginal applicants and medical students and at supporting their journeys towards the attainment of medical degrees. More research is needed to evaluate these initiatives' effectiveness. The results of such studies would not only provide needed information aimed at meeting the specific health needs of Aboriginal people, but may also contribute towards the laying of a framework to help narrow the gaps that exist within health care delivery to other minority groups. PMID- 16262806 TI - Competency-based assessment and cultural compression in medical education: lessons from educational anthropology. AB - OBJECTIVE: This paper explores the thesis that medical education is the cultural transmission to learners of specific values, which are increasingly expressed as graduation competencies. As testing is a powerful way to transmit cultural values to learners in a brief period of time, competency-based assessments can be an instrument of cultural compression in medical education. METHODS: The author reviewed medical literature to illustrate the concepts from educational anthropology, led the process one medical school used to develop its list of graduation competencies, and conducted a citation search about competency domains. RESULTS: There is support in the literature for viewing medical education as an example of cultural transmission and compression and for the assertion that testing influences student behaviour. The graduation competency statements developed by the school reflect traditional and emergent values. The citation search data confirmed that some competency domains reflected traditional values, while others reflected more emergent values. CONCLUSION: Concepts from educational anthropology are relevant to medical education and provide perspectives for understanding contemporary issues such as competency-based assessments. PMID- 16262807 TI - Integrating gender into a basic medical curriculum. AB - INTRODUCTION: In 1998, gaps were found to exist in the basic medical curriculum of the Radboud University Nijmegen Medical Centre regarding health-related gender differences in terms of biological, psychological and social factors. After screening the curriculum for language, content and context, adjustments aimed at incorporating gender issues were proposed. The aim of this study was to evaluate those adjustments, as well as to investigate whether gender had been successfully incorporated into the basic medical curriculum, and to identify the factors that played a role in this. METHODS: The education material of 9 curricular blocks was re-evaluated and interviews were held with block co-ordinators. RESULTS: Since the beginning of the project, gender has increasingly been brought to the attention of the students. Various factors have played a role: concrete and directly executable content-oriented proposals for adjustment; adequate translation of gender differences into actual patient care; motivated block co ordinators; the presence of a 'trigger person' in the faculty; incorporation into the existing education programme; the involvement of block co-ordinators in decision making, and the provision of practical support. DISCUSSION: Integrating gender into the basic medical curriculum has been largely successful. Block co ordinators' personal recognition of the importance of gender in patient care greatly facilitated implementation. The evaluation stimulated the forming of new ideas. It is recommended that these factors and those mentioned above should be taken into consideration when integrating gender into other faculties. PMID- 16262808 TI - Effect of ethnicity and gender on performance in undergraduate medical examinations. AB - OBJECTIVE: To assess the effect of ethnicity and gender on medical student examination performance. DESIGN: Cohort study of Year 3 medical students in 2002 and 2003. SETTING: Royal Free and University College Medical School, Imperial College School of Medicine. SUBJECTS: A total of 1216 Year 3 medical students, of whom 528 were male and 688 female, and 737 were white European and 479 Asian. OUTCOME MEASURE: Performance in summative written and objective structured clinical examinations (OSCEs) in July 2002 and 2003. RESULTS: White females performed best in all OSCEs and in 3 out of 4 written examinations. Mean scores for each OSCE and 2 out of 4 written examinations were higher for white students than for Asian students. The overall size of the effect is relatively small, being around 1-2%. CONCLUSION: Students of Asian origin, of both genders, educated in the UK, using English as their first language, continue to perform less well in OSCEs and written assessments than their white European peers. PMID- 16262809 TI - Consultant attitudes to undertaking undergraduate teaching duties: perspectives from hospitals serving a large medical school. AB - OBJECTIVE: To explore attitudes among National Health Service consultants responsible for delivering basic clinical teaching to medical students. DESIGN: Postal questionnaire. SUBJECTS AND SETTING: A total of 308 acute hospital trust consultants working in 4 'new' and 4 'established' teaching hospitals in the West Midlands metropolitan area, and involved in the delivery of clinical teaching to Year 3 medical students at the University of Birmingham Medical School during 2002-03. MAIN OUTCOME MEASURE(S): The questionnaire explored contractual requirements, actual teaching commitments and perceptions of medical students' knowledge and attitudes. Responses from doctors and surgeons and from respondents working in established and new teaching hospitals were compared. RESULTS: A total of 249 responses were received (response rate 80.8%). Although many consultants enjoy teaching students, their enjoyment and their ability to deliver high standards of teaching are compromised by time and resource constraints. For many the situation is aggravated by the perceived inappropriate organisation of the clinical teaching curriculum and the inadequate preparation of students for clinical practice. Linking these themes is the overarching perception among teachers that neither service nor educational establishments afford teaching the levels of recognition and reward associated with clinical work or research. CONCLUSION: To overcome barriers to teaching requires more reciprocal links between hospital staff and medical schools, opportunities for consultants to understand and to comment on curricular and timetable developments, and, perhaps most importantly, recognition (in contractual, financial, managerial and personal terms) of the importance of undergraduate teaching in the competing triad of service, research and education. PMID- 16262810 TI - Response to 'explanations in consultations: the combined effectiveness of doctors' and nurses' communication with patients'. PMID- 16262811 TI - The definition of 'portfolio'. PMID- 16262815 TI - A new evaluation tool for admissions. PMID- 16262816 TI - Introduction of problem-solving activities during conventional lectures. PMID- 16262817 TI - Student perceptions of undergraduate teachers. PMID- 16262818 TI - Horizontal integration in teaching within a biomedical department. PMID- 16262819 TI - Integration of ultrasound in the education programme in anatomy. PMID- 16262820 TI - Wound closure: an instructional DVD. PMID- 16262821 TI - Randomised trial of early clinical training for students. PMID- 16262822 TI - Physical examination teaching curriculum for senior medical students. PMID- 16262823 TI - Online classrooms enhance clerkship small group teaching. PMID- 16262824 TI - A systematic approach for teaching the home visit. PMID- 16262826 TI - Simulation training for medical emergencies in general practice. PMID- 16262827 TI - A new simulator-based psychological training on crisis management. PMID- 16262825 TI - Virtual patients get real. PMID- 16262829 TI - Can peer review help the marking experience? PMID- 16262828 TI - Video improved role play for teaching communication skills. PMID- 16262830 TI - Teaching communication skills for handover: perioperative specialist practitioners. PMID- 16262831 TI - Healing and hopefulness: a tool for doctor well-being. PMID- 16262832 TI - Medical education fellowship: fostering early interest in education. PMID- 16262833 TI - ROMP & ROL: peer-based professional development. PMID- 16262834 TI - Using an objective structured teaching evaluation for faculty development. PMID- 16262835 TI - Global health equity as the focus of graduate medical education. PMID- 16262836 TI - Peace-building through an international child health elective. PMID- 16262837 TI - Teaching chronic disease management to interdisciplinary medical learners. PMID- 16262838 TI - An electrocardiogram curriculum for resident doctors. PMID- 16262839 TI - Centralising curriculum feedback from graduates of small programmes. PMID- 16262840 TI - Mission Statement Day: introducing professionalism to medical students. PMID- 16262842 TI - Does student teaching harm or help cancer patients? PMID- 16262841 TI - Promoting humanistic care in the house staff continuity clinic. PMID- 16262843 TI - The Healer's Art: professionalism, service and mission. PMID- 16262844 TI - An effective tool for feedback on a problem-based learning (PBL) course. PMID- 16262846 TI - Portfolio assessment using a structured interview. PMID- 16262845 TI - Web-based diaries--windows to student internship feedback experiences. PMID- 16262847 TI - Review of capture-recapture methods applicable to noninvasive genetic sampling. AB - The use of noninvasive genetic sampling to identify individual animals for capture-recapture studies has become widespread in the past decade. Strong emphasis has been placed on the field protocols and genetic analyses with fruitful results. Little attention has been paid to the capture-recapture application for this specific type of data beyond stating the effects of assumption violations. Here, we review the broad class of capture-recapture methods that are available for use with DNA-based capture-recapture data, noting the array of biologically interesting parameters such as survival, emigration rates, state transition rates and the finite rate of population change that can be estimated from such data. We highlight recent developments in capture recapture theory specifically designed for noninvasive genetic sampling data. PMID- 16262848 TI - Recent invasion of the tropical Atlantic by an Indo-Pacific coral reef fish. AB - The last tropical connection between Atlantic and Indian-Pacific habitats closed c. 2 million years ago (Ma), with the onset of cold-water upwelling off southwestern Africa. Yet comparative morphology indicates more recent connections in several taxa, including reef-associated gobies (genus Gnatholepis). Coalescence and phylogenetic analyses of mtDNA cytochrome b sequences demonstrate that Gnatholepis invaded the Atlantic during an interglacial period approximately 145,000 years ago (d = 0.0054), colonizing from the Indian Ocean to the western Atlantic, and subsequently to the central ( approximately 100,000 years ago) and eastern Atlantic ( approximately 30,000 years ago). Census data show a contemporary range expansion in the northeastern Atlantic linked to global warming. PMID- 16262849 TI - Wide-range analysis of genetic structure of Betula maximowicziana, a long-lived pioneer tree species and noble hardwood in the cool temperate zone of Japan. AB - Betula maximowicziana is a long-lived pioneer tree species in Japanese cool temperate forests that plays an important role in maintenance of the forest ecosystem and has high economic value. Here we assess the wide-range genetic structure of 23 natural populations of B. maximowicziana using 11 simple sequence repeat (SSR) loci. Genetic diversity within populations was relatively low in all populations (mean H(E), 0.361; mean allelic richness, 2.80; mean rare allelic richness, 1.02). The population differentiation was also relatively low (F(ST), 0.062). Genetic distance-based and Bayesian clustering analysis revealed that the populations examined here could be divided into a southern group and a northern group. Analysis of rare allelic richness and Bayesian clustering revealed evidence for both southern and northern refugia during the last glacial period. Furthermore, a comparison of regional genetic diversity revealed significant clines in allelic richness. In spatial genetic structure evaluation, significant isolation by distance (IBD) was detected among the 23 populations, but not within regions. Moreover, significant population bottlenecks were found in all populations under infinite allele model (IAM) assumptions. These unusual, significant bottlenecks might be because of the processes of postglacial colonization and the species' characters and/or life history as a long-lived pioneer tree species. The wide-range, regional genetic structure found in this study provides an important baseline for conservation and forest management, including the identification of evolutionarily significant units (ESUs) and/or management units (MUs) of B. maximowicziana. PMID- 16262850 TI - Colonization and dispersal in a social species, the Bechstein's bat (Myotis bechsteinii). AB - Metapopulation genetic models consider that colonization and dispersal are distinct behaviours. However, whether colonization and dispersal indeed reflect different biological processes in nature is unclear. One possibility to test this assumption is to assess patterns of autosomal and mitochondrial genetic structure in species with strict female philopatry, such as the communally breeding Bechstein's bat. In this species, mitochondrial DNA can spread only when females establish new colonies, and autosomal DNA is transmitted among colonies only when females mate with solitary males born in foreign colonies. Investigating the genetic structure among 37 colonies, we found that autosomal genes followed an island model on a regional scale and a model of isolation by distance on a larger geographical scale. In contrast, mitochondrial genetic structure revealed no pattern of isolation by distance at a large scale but exhibited an effect of ecological barriers on a regional scale. Our results provide strong empirical evidence that colonization and dispersal do not follow the same behavioural rules in this bat, supporting the assumption of metapopulation genetic models. PMID- 16262851 TI - Native breeds demonstrate high contributions to the molecular variation in northern European sheep. AB - Population contribution to genetic diversity can be estimated using neutral variation. However, population expansion or hybridization of diverged ancestries may weaken correlation between neutral and non-neutral variation. Microsatellite variation was studied at 25 loci in 20 native and 12 modern or imported northern European sheep breeds. Breed contributions to total gene diversity, allelic richness and mean allele-sharing distance between individuals were measured. Indications of changes in population size and admixtures of divergent ancestries were investigated and the extent of inbreeding was estimated. The northern European sheep demonstrated signs of reduction in effective population size. Many old, small populations made a substantial positive contribution to total molecular variation, but populations with several divergent major ancestries did not contribute substantially to molecular variation, with the exception of the Norwegian Rygja sheep. However, several diverged major ancestries may cause it to contribute less to non-neutral variation than expected from the microsatellite data. Breed uniqueness and within-breed variability generally had opposite effects on breed contributions to molecular diversity. The degree of inbreeding did not reflect the breed contribution to total gene diversity or allelic richness, but inbred populations increased the mean allele-sharing distance between individuals. Our study indicates breed conservation to be especially important in maintaining allelic variation in northern European sheep and supports the evolutionary importance of peripheral populations. PMID- 16262852 TI - Population genetic analysis identifies source-sink dynamics for two sympatric garter snake species (Thamnophis elegans and Thamnophis sirtalis). AB - Population genetic structure can be shaped by multiple ecological and evolutionary factors, but the genetic consequences of these factors for multiple species inhabiting the same environment remain unexplored. We used microsatellite markers to examine the population structures of two coexisting species of garter snake, Thamnophis elegans and Thamnophis sirtalis, to determine if shared landscape and biology imposed similar population genetic structures. These snakes inhabit a series of ponds, lakes and flooded meadows in northern California and tend to converge on prey type wherever they coexist. Both garter snakes had comparable effective population sizes and bidirectional migration rates (estimated using a maximum-likelihood method based on the coalescent) with low but significant levels of genetic differentiation (F(ST) = 0.024 for T. elegans and 0.035 for T. sirtalis). Asymmetrical gene flow revealed large source populations for both species as well as potential sinks, suggesting frequent extinction-recolonization and metapopulation dynamics. In addition, we found a significant correlation between their genetic structures based on both pairwise F(ST)s for shared populations (P = 0.009) and for bidirectional migration rates (P = 0.024). Possible ecological and evolutionary factors influencing similarities and differences in genetic structure for the two species are discussed. Genetic measures of effective population size and migration rates obtained in this study are also compared with estimates obtained from mark recapture data. PMID- 16262853 TI - Molecular evidence for host specificity of parasitic nematode microfilariae in some African rainforest birds. AB - Here we describe, determine the prevalence, and examine the host-specificity of some parasitic nematode microfilariae in selected bird species from West and Central Africa. We used microscopy to determine the prevalence of microfilariae in 969 host individuals representing 121 rainforest bird species from Cameroon, Cote d'Ivoire and Equatorial Guinea. Thirteen (11%) of these potential host species harboured microfilariae, and 35 individuals (3.6%) were infected. From the 35 infected individuals, we identified eight distinct morphological microfilarial forms. Sixteen of the 35 infected individuals were of one host species, the Fire-crested Alethe (Alethe diademata), at a prevalence rate of 62%. To examine host and geographical specificity, we sequenced a portion of the LSU rDNA gene from representative microfilariae drawn from different hosts and collecting locations. Identical sequences of the nematode LSU rDNA gene were found in A. diademata collected from locations in Cote d'Ivoire and Equatorial Guinea, locations separated by the Dahomey Gap and associated with different hypothesized refugial areas. In contrast, several other bird species collected at the same sites harboured different microfilaria lineages. We sequenced the mitochondrial ATP synthase genes of the host species A. diademata, and found a 5.4% sequence divergence between the birds sampled in Cote d'Ivoire, and those from Cameroon. Thus, despite this split between the two populations, they harbour microfilariae with identical lineages. These data provide evidence that the microfilariae found in A. diademata may be highly host specific. This apparent specificity may have important implications for the evolutionary and ecological interactions between parasitic nematodes and their avian hosts. PMID- 16262855 TI - Parallels in the evolution of the two largest New and Old World seed-beetle genera (Coleoptera, Bruchidae). AB - This study provides the first phylogenetic analysis of a large sample of the two largest genera of seed-beetles, Acanthoscelides Schilsky and Bruchidius Schilsky, which mostly feed on legumes (Fabaceae). The goal of this study was to investigate evolutionary patterns in relation to biogeography and host-plant associations. We used three mitochondrial molecular markers and parsimony and Bayesian inference methods to reconstruct the phylogeny of 76 species. In addition, we critically reviewed host-plant records in the literature for these two bruchid genera. Our results demonstrated the existence of two major clades, one New World and one largely Old World, which generally correspond to the two genera. Yet, current classification of several species is erroneous, so that both genera as currently defined are paraphyletic. We highlighted a strong trend toward specialization (with high taxonomic conservatism in host-plant use) exhibited by the two studied genera. However, we showed the existence of several host shifts during the evolution of this group of bruchids. Our phylogenetic hypotheses and our evaluation of host-plant associations both suggest that the two genera have undergone parallel evolution, as they have independently colonized similar host plants in their respective areas of distribution. Our estimation of divergence times indicated a more ancient origin for bruchids than that suggested by the fossil records. Interestingly, the suggested timing of diversification is consistent with the hypothesis of a radiation that could have occurred contemporaneously with the diversification of their legume hosts. PMID- 16262854 TI - Phylogeography of the black fly Simulium tani (Diptera: Simuliidae) from Thailand as inferred from mtDNA sequences. AB - Intraspecific phylogeography has been used widely as a tool to infer population history. However, little attention has been paid to Southeast Asia despite its importance in terms of biodiversity. Here we used the cytochrome oxidase I gene of mitochondrial DNA (mtDNA) for a phylogeographic study of 147 individuals of the black fly Simulium tani from Thailand. The mtDNA revealed high genetic differentiation between the major geographical regions of north, east and central/south Thailand. Mismatch distributions indicate population expansions during the mid-Pleistocene and the late Pleistocene suggesting that current population structure and diversity may be due in part to the species' response to Pleistocene climatic fluctuations. The genealogical structure of the haplotypes, high northern diversity and maximum-likelihood inference of historical migration rates, suggest that the eastern and central/southern populations originated from northern populations in the mid-Pleistocene. Subsequently, the eastern region had had a largely independent history but the central/southern population may be largely the result of recent (c. 100,000 years ago) expansion, either from the north again, or from a relictual population in the central region. Cytological investigation revealed that populations from the south and east have two overlapping fixed chromosomal inversions. Since these populations also share ecological characteristics it suggests that inversions are involved in ecological adaptation. In conclusion both contemporary and historical ecological conditions are playing an important role in determining population genetic structure and diversity. PMID- 16262857 TI - Evolutionary genetics and biogeographic structure of Rhizobium gallicum sensu lato, a widely distributed bacterial symbiont of diverse legumes. AB - We used phylogenetic and population genetics approaches to evaluate the importance of the evolutionary forces on shaping the genetic structure of Rhizobium gallicum and related species. We analysed 54 strains from several populations distributed in the Northern Hemisphere, using nucleotide sequences of three 'core' chromosomal genes (rrs, glnII and atpD) and two 'auxiliary' symbiotic genes (nifH and nodB) to elucidate the biogeographic history of the species and symbiotic ecotypes (biovarieties) within species. The analyses revealed that strains classified as Rhizobium mongolense and Rhizobium yanglingense belong to the chromosomal evolutionary lineage of R. gallicum and harbour symbiotic genes corresponding to a new biovar; we propose their reclassification as R. gallicum bv. orientale. The comparison of the chromosomal and symbiotic genes revealed evidence of lateral transfer of symbiotic information within and across species. Genetic differentiation analyses based on the chromosomal protein-coding genes revealed a biogeographic pattern with three main populations, whereas the 16S rDNA sequences did not resolve that biogeographic pattern. Both the phylogenetic and population genetic analyses showed evidence of recombination at the rrs locus. We discuss our results in the light of the contrasting views of bacterial species expressed by microbial taxonomist and evolutionary biologists. PMID- 16262856 TI - Importance of genetic drift during Pleistocene divergence as revealed by analyses of genomic variation. AB - Determining what factors affect the structuring of genetic variation is key to deciphering the relative roles of different evolutionary processes in species differentiation. Such information is especially critical to understanding how the frequent shifts and fragmentation of species distributions during the Pleistocene translates into species differences, and why the effect of such rapid climate change on patterns of species diversity varies among taxa. Studies of mitochondrial DNA (mtDNA) have detected significant population structure in many species, including those directly impacted by the glacial cycles. Yet, understanding the ultimate consequence of such structure, as it relates to how species divergence occurs, requires demonstration that such patterns are also shared with genomic patterns of differentiation. Here we present analyses of amplified fragment length polymorphisms (AFLPs) in the montane grasshopper Melanoplus oregonensis to assess the evolutionary significance of past demographic events and associated drift-induced divergence as inferred from mtDNA. As an inhabitant of the sky islands of the northern Rocky Mountains, this species was subject to repeated and frequent shifts in species distribution in response to the many glacial cycles. Nevertheless, significant genetic structuring of M. oregonensis is evident at two different geographic and temporal scales: recent divergence associated with the recolonization of the montane meadows in individual sky islands, as well as older divergence associated with displacements into regional glacial refugia. The genomic analyses indicate that drift-induced divergence, despite the lack of long-standing geographic barriers, has significantly contributed to species divergence during the Pleistocene. Moreover, the finding that divergence associated with past demographic events involves the repartitioning of ancestral variation without significant reductions of genomic diversity has intriguing implications - namely, the further amplification of drift-induced divergence by selection. PMID- 16262858 TI - Historical colonization and demography of the Mediterranean damselfish, Chromis chromis. AB - The desiccation of the Mediterranean Sea during the Messinian Salinity Crisis 6.0 5.3 million years ago (Ma), caused a major extinction of the marine ichthyofauna of the Mediterranean. This was followed by an abrupt replenishment of the Mediterranean from the Atlantic after the opening of the Strait of Gibraltar. In this study, we combined demographic and phylogeographic approaches using mitochondrial and nuclear DNA markers to test the alternative hypotheses of where (Atlantic or Mediterranean) and when (before or after the Messinian Salinity Crisis) speciation occurred in the Mediterranean damselfish, Chromis chromis. The closely related geminate transisthmian pair Chromis multilineata and Chromis atrilobata was used as a way of obtaining an internally calibrated molecular clock. We estimated C. chromis speciation timing both by determining the time of divergence between C. chromis and its Atlantic sister species Chromis limbata (0.93-3.26 Ma depending on the molecular marker used, e.g. 1.23-1.39 Ma for the control region), and by determining the time of coalescence for C. chromis based on mitochondrial control region sequences (0.14-0.21 Ma). The time of speciation of C. chromis was always posterior to the replenishment of the Mediterranean basin, after the Messinian Salinity Crisis. Within the Mediterranean, C. chromis population structure and demographic characteristics revealed a genetic break at the Peloponnese, Greece, with directional and eastbound gene flow between western and eastern groups. The eastern group was found to be more recent and with a faster growing population (coalescent time = 0.09-0.13 Ma, growth = 485.3) than the western group (coalescent time = 0.13-0.20 Ma, growth = 325.6). Our data thus suggested a western origin of C. chromis, most likely within the Mediterranean. Low sea water levels during the glacial periods, the hydrographic regime of the Mediterranean and dispersal restriction during the short pelagic larval phase of C. chromis (18-19 days) have probably played an important role in C. chromis historical colonization. PMID- 16262859 TI - Evolutionary processes in a continental island system: molecular phylogeography of the Aegean Nigella arvensis alliance (Ranunculaceae) inferred from chloroplast DNA. AB - Continental shelf island systems, created by rising sea levels, provide a premier setting for studying the effects of past fragmentation, dispersal, and genetic drift on taxon diversification. We used phylogeographical (nested clade) and population genetic analyses to elucidate the relative roles of these processes in the evolutionary history of the Aegean Nigella arvensis alliance (= 'coenospecies'). We surveyed chloroplast DNA (cpDNA) variation in 455 individuals from 47 populations (nine taxa) of the alliance throughout its core range in the Aegean Archipelago and surrounding mainland areas of Greece and Turkey. The study revealed the presence of three major lineages, with largely nonoverlapping distributions in the Western, Central, and Eastern Aegean. There is evidence supporting the idea that these major lineages evolved in situ from a widespread (pan-Aegean) ancestral stock as a result of multiple fragmentation events, possibly due to the influence of post-Messinian sea flooding, Pleistocene eustatic changes and corresponding climate fluctuations. Over-sea dispersal and founder events appear to have played a rather insignificant role in the group's history. Rather, all analytical approaches identified the alliance as an organism group with poor seed dispersal capabilities and a susceptibility to genetic drift. In particular, we inferred that the observed level of cpDNA differentiation between Kikladian island populations of Nigella degenii largely reflects population history, (viz. Holocene island fragmentation) and genetic drift in the near absence of seed flow since their time of common ancestry. Overall, our cpDNA data for the N. arvensis alliance in general, and N. degenii in particular, indicate that historical events were important in determining the phylogeographical patterns seen, and that genetic drift has historically been relatively more influential on population structure than has cytoplasmic gene flow. PMID- 16262861 TI - Unbiased estimation of relative reproductive success of different groups: evaluation and correction of bias caused by parentage assignment errors. AB - Parentage assignment is widely applied to studies on mating systems, population dynamics and natural selection. However, little is known about the consequence of assignment errors, especially when some parents are not sampled. We investigated the effects of two types of error in parentage assignment, failing to assign a true parent (type A) and assigning an untrue parent (type B), on an estimate of the relative reproductive success (RRS) of two groups of parents. Employing a mathematical approach, we found that (i) when all parents are sampled, minimizing either type A or type B error insures the minimum bias on RRS, and (ii) when a large number of parents is not sampled, type B error substantially biases the estimated RRS towards one. Interestingly, however, (iii) when all parents were sampled and both error rates were moderately high, type A error biased the estimated RRS even more than type B error. We propose new methods to obtain an unbiased estimate of RRS and the number of offspring whose parents are not sampled (zW(z)), by correcting the error effects. Applying them to genotypic data from steelhead trout (Oncorhynchus mykiss), we illustrated how to estimate and control the assignment errors. In the data, we observed up to a 30% assignment error and a strong trade-off between the two types of error, depending on the stringency of the assignment decision criterion. We show that our methods can efficiently estimate an unbiased RRS and zW(z) regardless of assignment method, and how to maximize the statistical power to detect a difference in reproductive success between groups. PMID- 16262860 TI - Tempo and mode of speciation in the Baja California disjunct fish species Anisotremus davidsonii. AB - The Baja California region provides a natural setting for studying the early mechanisms of allopatric speciation in marine systems. Disjunct fish populations from several species that occur in the northern Gulf of California and northern Pacific coast of Baja California, but are absent from its southern shores, were previously shown to be genetically isolated, making them excellent candidates for studying allopatry. In addition, one of these species, the sargo Anisotremus davidsonii, has two pairs of congeneric Panamic trans-isthmian geminate species that allow for internal molecular clock calibration. Phylogeographic and demographic approaches based on mitochondrial (cytochrome b) and nuclear (S7 ribosomal protein) sequences showed that A. davidsonii entered the gulf from the south, and later colonized the Pacific coast, approximately 0.6-0.16 million years ago. Pacific coast colonization may have used a route either around the southern cape of Baja California or across the peninsula through a natural seaway. However, while several seaways have been described from different geological times, none matches the dates of population disjunction, yet much geological work remains to be done in that area. At the present time, there is no evidence for dispersal around the southern tip of the Baja California Peninsula. Signatures of incipient allopatric speciation were observed, such as the reciprocal monophyly of disjunct populations for the mitochondrial marker. However, other characteristics were lacking, such as a strong difference in divergence and coalescence times. Taken together, these results suggest that disjunct populations of A. davidsonii may be consistent with the earliest stages of allopatric speciation. PMID- 16262862 TI - The potential costs of accounting for genotypic errors in molecular parentage analyses. AB - Genotypic errors, whether due to mutation or laboratory error, can cause the genotypes of parents and their offspring to appear inconsistent with Mendelian inheritance. As a result, molecular parentage analyses are expected to benefit when allowances are made for the presence of genotypic errors. However, a cost of allowing for genotypic errors might also be expected under some analytical conditions, primarily because parentage analyses that assume nonzero genotypic error rates can neither assign nor exclude parentage with certainty. The goal of this work was therefore to determine whether or not such costs might be important under conditions relevant to parentage analyses, particularly in natural populations. Simulation results indicate that the costs may often outweigh the benefits of accounting for nonzero error rates, except in situations where data are available for many marker loci. Consequently, the most powerful approach to handling genotypic errors in parentage analyses might be to apply likelihood equations with error rates set to values substantially lower than the rates at which genotypic errors occur. When applying molecular parentage analyses to natural populations, we advocate an increased consideration of optimal strategies for handling genotypic errors. Currently available software packages contain procedures that can be used for this purpose. PMID- 16262863 TI - It's good to be queen: classically eusocial colony structure and low worker fitness in an obligately social sweat bee. AB - Lasioglossum malachurum, a bee species common across much of Europe, is obligately eusocial across its range but exhibits clear geographic variation in demography and social behaviour. This variation suggests that social interactions between queens and workers, opportunities for worker oviposition, and patterns of relatedness among nest mates may vary considerably, both within and among regions. In this study, we used three microsatellite loci with 12-18 alleles each to examine the sociogenetic structure of colonies from a population at Agios Nikolaos Monemvasias in southern Greece. These analyses reveal that the majority of colonies exhibit classical eusocial colony structure in which a single queen mated to a single male monopolizes oviposition. Nevertheless, we also detect low rates of multiqueen nest founding, occasional caste switching by worker-destined females, and worker oviposition of both gyne and male-producing eggs in the final brood. Previous evidence that the majority of workers show some ovarian development and a minority (17%) have at least one large oocyte contrasts with the observation that only 2-3% of gynes and males (the so-called reproductive brood) are produced by workers. An evaluation of the parameters of Hamilton's Rule suggests that queens benefit greatly from the help provided by workers but that workers achieve greater fitness by provisioning and laying their own eggs rather than by tending to the queen's eggs. This conflict of interest between the queen and her workers suggests that the discrepancy between potential and achieved worker oviposition is due to queen interference. Comparison of relatedness and maternity patterns in the Agios Nikolaos Monemvasias population with those from a northern population near Tubingen, Germany, points to a north south cline of increasingly effective queen control of worker behaviour. PMID- 16262864 TI - Nitrogen-fixing nodules from rose wood legume trees (Dalbergia spp.) endemic to Madagascar host seven different genera belonging to alpha- and beta Proteobacteria. AB - Although legume biodiversity is concentrated in tropical regions, the majority of studies on legume nodulating bacteria (LNB) are focused on cultivated leguminous plants from temperate regions. However, recent works on tropical regions tend to indicate that the actual diversity of LNB is largely underestimated. In this study, we report the isolation and characterization of 68 nitrogen-fixing root nodule bacteria collected from eight endemic tree species of Dalbergia in Madagascar. The isolates were characterized by (i) restriction fragment length polymorphism (RFLP) analysis of 16S-IGS rDNA, (ii) 16S rDNA gene sequencing and (iii) nodulation tests. Results revealed a wide diversity of bacteria present in the nodules of Dalbergia. Among the 68 isolated bacteria, 65 belonged to Bradyrhizobium, Mesorhizobium, Rhizobium, Azorhizobium and Phyllobacterium from the alpha-class of Proteobacteria, and three isolates belonged to Burkholderia and Ralstonia from the beta-class of Proteobacteria. Our results also show for the first time that a strain belonging to the Burkholderia cepacia complex is able to induce efficient nodules on a legume plant. PMID- 16262866 TI - Resistance to abomasal nematodes and individual genetic variability in reindeer. AB - Resistance to parasites is believed to have a widespread influence on demographic and adaptive processes. In systems where parasites impose a fitness cost on their host, heterozygotes may be selected because they are more resistant to parasites than homozygotes. Our objective was to assess the relationships between genomewide individual heterozygosity and abomasal nematode burdens in female Svalbard reindeer (Rangifer tarandus platyrhynchus) after the effects of host age, locality, season, and year had been accounted for. Samples were obtained from 306 female reindeer that were culled and genotyped at nine microsatellite loci. Reindeer in our study populations are mainly parasitized by the gastrointestinal nematodes Ostertagia gruehneri and Marshallagia marshalli. The infection intensity of each parasite differed between subpopulations, and among host age classes, seasons and years. We found no significant relationships between abomasal worm burdens, or lumen and mucosa larvae, of either O. gruehneri or M. marshalli and individual heterozygosity (or mean d(2)) alone or in interactions with host age, locality, and year. Although we analysed one of the largest data set available to date on gastrointestinal nematodes of a wild ruminant, we used a typical data set of nine genetic neutral markers that may have had low power to detect heterozygosity-fitness correlations. We conclude that the proportion of the variance in parasite resistance explained by individual heterozygosity for neutral genetic markers is low in Svalbard reindeer and in vertebrates in general, and we suggest that the candidate-gene approach might be more fruitful for further research on gene-fitness correlations. PMID- 16262865 TI - The significance of facultative scavenging in generalist predator nutrition: detecting decayed prey in the guts of predators using PCR. AB - Gut-content analyses using molecular techniques are an effective approach to quantifying predator-prey interactions. Predation is often assumed but scavenging is an equally likely route by which animal DNA enters the gut of a predator/scavenger. We used PCR (polymerase chain reaction) to detect scavenged material in predator gut homogenates. The rates at which DNA in decaying slugs (Mollusca: Pulmonata) and aphids (Homoptera: Aphididae) became undetectable were estimated. The detectability of DNA from both carrion types in the guts of the generalist predator Pterostichus melanarius (Coleoptera: Carabidae) was then determined. The effects of carrion age and weight, as well as beetle sex, on detection periods, were quantified. Laboratory trials measured prey preference of beetles between live and decaying prey. Further experiments measured, for the first time, feeding by P. melanarius on dead slugs and aphids directly in the field. In both field and laboratory, P. melanarius preferentially fed on dead prey if available, but preference changed as the prey became increasingly decayed. Disappearance rates for slug carrion in wheat fields and grasslands were estimated and P. melanarius was identified as the main scavenger. Comparison of the retention time for dead slugs in the field, with the detection period for decaying slug material in the guts of the predators, showed that PCR-based techniques are not able to distinguish between predated and scavenged food items. This could potentially lead to overestimation of the impact of predation on slugs (and other prey) by carabids. Possible implications of facultative scavenging by invertebrate predators for biocontrol and food-web research are discussed. PMID- 16262867 TI - Temporal patterns of genetic variation across a 9-year-old aerial seed bank of the shrub Banksia hookeriana (Proteaceae). AB - The pattern of accumulation of genetic variation over time in seed banks is poorly understood. We examined the genetic structure of the aerial seed bank of Banksia hookeriana within a single 15-year-old population in fire-prone southwestern Australia, and compared genetic variation between adults and each year of a 9-year-old seed bank using amplified fragment length polymorphism (AFLP). B. hookeriana is well suited to the study of seed bank dynamics due to the canopy storage of its seeds, and because each annual crop can be identified. A total of 304 seeds from nine crop years and five maternal plants were genotyped, along with 113 plants from the adult population. Genetic variation, as assessed by the proportion of polymorphic markers (P(p)) and Shannon's index (I), increased slightly within the seed bank over time, while gene diversity (H(j)), did not change. P(p), I, and H(j) all indicated that genetic variation within the seed bank quickly approached the maximal level detected. Analysis of molecular variance revealed that less than 4% of variation could be accounted for by variation among seeds produced in different years, whereas there was greater differentiation among maternal plants (12.7%), and among individual seeds produced by different maternal plants (83.4%). With increasing population age, offspring generated each year were slightly more outbred, as indicated by an increase in the mean number of nonmaternal markers per offspring. There were no significant differences for H(j) or I between adults and the seed bank. Viability of seeds decreased with age, such that the viability of 9-year-old seeds was half that of 2-year-old seeds. These results suggest that variable fire frequencies have only limited potential to influence the amount of genetic variation stored within the seed bank of B. hookeriana. PMID- 16262868 TI - Variation at two flowering time genes within and among populations of Arabidopsis thaliana: comparison with markers and traits. AB - Flowering Locus C (FLC) and Frigida are two interacting genes controlling flowering time variation in Arabidopsis thaliana. Variation at these genes was surveyed in 12 A. thaliana populations sampled in France. These populations were also screened for variation at molecular markers [12 microsatellites and 19 cleaved amplified polymorphic sequence (CAPS) markers] and at seven quantitative traits measured with and without vernalization. Seven populations were highly polymorphic at markers (H(S) = 0.57 at microsatellites, 0.24 at CAPS) and showed heritable variation for bolting time and some other traits. Five populations were genetically fixed or nearly fixed. Q(ST) for bolting time without vernalization was significantly higher than F(ST), suggesting local divergent selection. One of the two haplotype groups at FLC (FLC(A)) was very predominant (frequency of 99%). The first exon of Frigida showed elevated nonsynonymous variation, and nine loss of-function mutations were found throughout the gene. The association between loss-of-function and earlier bolting was confirmed. Overall, 18 Frigida haplotypes were detected. The pattern of variation at Frigida was largely similar to that found at markers and traits, with the same populations being fixed or highly diverse. Metapopulation dynamics is thus probably the main factor shaping genetic variation in A. thaliana. However, F(ST) for functional (FRI) vs. nonfunctional (FRI(Delta)) haplotypes was significantly higher than F(ST) at markers. This suggested that loss-of-function at Frigida is under local selection for flowering time. PMID- 16262869 TI - Use of sequence data from rainbow trout and Atlantic salmon for SNP detection in Pacific salmon. AB - Single nucleotide polymorphisms (SNPs) are a class of genetic markers that are well suited to a broad range of research and management applications. Although advances in genotyping chemistries and analysis methods continue to increase the potential advantages of using SNPs to address molecular ecological questions, the scarcity of available DNA sequence data for most species has limited marker development. As the number and diversity of species being targeted for large scale sequencing has increased, so has the potential for using sequence from sister taxa for marker development in species of interest. We evaluated the use of Oncorhynchus mykiss and Salmo salar sequence data to identify SNPs in three other species (Oncorhynchus tshawytscha, Oncorhynchus nerka and Oncorhynchus keta). Primers designed based on O. mykiss and S. salar alignments were more successful than primers designed based on Oncorhynchus-only alignments for sequencing target species, presumably due to the much larger number of potential targets available from the former alignments and possibly greater sequence conservation in those targets. In sequencing approximately 89 kb we observed a frequency of 4.30 x 10(-3) SNPs per base pair. Approximately half (53/101) of the subsequently designed validation assays resulted in high-throughput SNP genotyping markers. We speculate that this relatively low conversion rate may reflect the duplicated nature of the salmon genome. Our results suggest that a large number of SNPs could be developed for Pacific salmon using sequence data from other species. While the costs of DNA sequencing are still significant, these must be compared to the costs of using other marker classes for a given application. PMID- 16262871 TI - Hydrolytic enzymes as virulence factors of Candida albicans. AB - Candida albicans is a facultative pathogenic micro-organism that has developed several virulence traits enabling invasion of host tissues and avoidance of host defence mechanisms. Virulence factors that contribute to this process are the hydrolytic enzymes. Most of them are extracellularly secreted by the fungus. The most discussed hydrolytic enzymes produced by C. albicans are secreted aspartic proteinases (Saps). The role of these Saps for C. albicans infections was carefully evaluated in numerous studies, whereas only little is known about the physiological role of the secreted phospholipases (PL) and almost nothing about the involvement of lipases (Lip) in virulence. They may play an important role in the pathogenicity of candidosis and their hydrolytic activity probably has a number of possible functions in addition to the simple role of digesting molecules for nutrition. Saps as the best-studied member of this group of hydrolytic enzymes contribute to host tissue invasion by digesting or destroying cell membranes and by degrading host surface molecules. There is also some evidence that hydrolytic enzymes are able to attack cells and molecules of the host immune system to avoid or resist antimicrobial activity. High hydrolytic activity with broad substrate specificity has been found in several Candida species, most notably in C. albicans. This activity is attributed to multigene families with at least 10 members for Saps and Lips and several members for PL B. Distinct members of these gene families are differentially regulated in various Candida infections. In future, prevention and control of Candida infections might be achieved by pharmacological or immunological tools specifically modulated to inhibit virulence factors, e.g. the family of Saps. PMID- 16262872 TI - Four cases of Candida albicans infections with isolates developing pink colonies on CHROMagar Candida plates. AB - Candida albicans, the most commonly isolated yeast species, is typically identified by its green colony-colour on CHROMagar Candida plates. We here report four cases of Candida albicans infections, in which the initial identification was non-albicans isolates due to a clear pink colour of the colonies on CHROMagar Candida plates. However, classical phenotypic criteria, biochemical assimilation pattern and molecular characterisation identified all four isolates as C. albicans isolates. PMID- 16262873 TI - Majocchi's granuloma of the vulva caused by Trichophyton mentagrophytes. AB - We report a case of Majocchi's granuloma caused by Trichophyton mentagrophytes on the vulva in a 23-year-old girl who had used topical steroids for many years. Her dog was a source of the infection. PMID- 16262874 TI - Susceptibility to antifungal agents of Candida species isolated from paediatric and adult patients with haematological diseases. AB - Summary The susceptibility to six antifungals: amphotericin B (AMF), 5 fluorocytosine (5-F), miconazole (MIK), ketoconazole (KET), fluconazole (FLU) and itraconazole (ITR) was tested among 206 Candida spp. isolated from paediatric and adult patients with haematological malignancies. To determinate the susceptibility the commercial microdilution method Fungitest (Bio-Rad, France) was used. The strains were classified as susceptible, intermediate susceptible, or resistant on the base of the growth in following breakpoint concentrations of particular drugs: 2 and 8 microg ml(-1) for AMF, 2 and 32 microg ml(-1) for 5-F, 0.5 and 8 microg ml(-1) for MIK, 0.5 and 4 microg ml(-1) for KET and ITR, and 8 and 64 microg ml(-1) for FLU. The highest activity to overall species showed AMF (only one resistant strain) and 5-F (85% susceptible strains). Most of C. albicans isolates were susceptible to tested azoles. The percentages of C. albicans resistant to FLU, ITR, KET and MIK were 4, 11, 8, and 0.8%, respectively. The less susceptible to azoles were C. glabrata and C. krusei (14% and 44% isolates resistant to FLU). A non-albicans Candida isolated from adult patients receiving KET prophylaxis was more frequently resistant to FLU than isolates from patients without previous exposure to azoles (P < 0.05). We did not observe differences in the susceptibility of Candida strains isolated from children compared with those from adults. PMID- 16262876 TI - Cerebral zygomycosis. AB - Fifty-six patients with cerebral zygomycosis (mucormycosis) were seen during the period 1971-2001 in two tertiary care hospitals located in south India with tropical climate and catering to neurological diseases. Forty-four patients had rhinocerebral and twelve patients had isolated central nervous system (CNS) zygomycosis. Of these, ten were culture proven (Rhizopus oryzae in eight and Mucor in two); 30 were diagnosed as probable and 16 were diagnosed possible; mixed infections were seen in three patients. Diabetes mellitus was the predisposing condition in a majority (31/44) of patients with the rhinocerebral form of zygomycosis. The tissue obtained at biopsy/autopsy in either form showed necrotic/infarcted tissue with neutrophilic infiltration with broad non-septate hyphae showing irregular branching. The outcome was poor despite surgical excision and antifungal therapy. The high concentration of spores in a mouldy environment, the bird population and improper disposal of hospital waste may facilitate healthy hosts presenting with primary CNS disease. PMID- 16262875 TI - The serum levels of calcium, magnesium, iron and zinc in patients with recurrent vulvovaginal candidosis during attack, remission and in healthy controls. AB - The real cause of recurrent vulvovaginal candidosis (RVVC) is concealed and the etiopathogenesis of this disease remains to be determined. In a cohort study, concentrations of metals in 44 patients with RVVC and 30 healthy age-matched women were measured and compared. The concentrations of serum calcium (Ca), magnesium (Mg) and iron (Fe) were measured photometrically, the zinc (Zn) levels were determined using flame atomic absorption spectrometry. For statistical analysis were used the Student's t-tests (paired analysis for attack vs. remission; non-paired analysis for patient vs. control). Although all measured metals were within normal ranges the patients with RVVC had in contrast to the healthy controls significantly lower levels of serum Ca, Mg and Zn and insignificantly higher levels of Fe. These relative changes may contribute to the development of attacks in patients with RVVC. PMID- 16262877 TI - Subcutaneous alternariosis. AB - A case of subcutaneous alternariosis caused by Alternaria alternata is reported in a 52-year-old male who underwent kidney transplantation 1 year earlier. The infection manifested as a vegetating mass about 7 cm in diameter on the right forearm. Histological examination with Grocott staining and periodic acid-Schiff diastase revealed fungal spores and hyphae with chronic granulomatous inflammation. Biopsy fragments inoculated on Sabouraud-glucose agar with chloramphenicol produced colonies which were identified as Alternaria alternata on the basis of macroscopic and microscopic characters. PMID- 16262878 TI - The causative pathogens of onychomycosis in southern Taiwan. AB - Selecting an appropriate antimycotic targeting the pathogens are among the most important factors for successfully treating onychomycosis. The aim of this study was to investigate the pathogens of onychomycosis in southern Taiwan and analyse the association between various factors and the distribution of pathogens. A total of 375 patients with onychomycosis were enrolled. Histopathological examination and fungus culture of nail specimens were performed. The pathogens were dermatophytes in 227 patients (60.5%), Candida in 118 (31.5%) and moulds in 30 (8%). Compared to patients with toenail involvement, the odds ratio (OR) for those with fingernail involvement to have non-dermatophytic onychomycosis (NDO), i.e. onychomycosis caused by Candida and moulds, was 5.04 [95% confidence interval (CI): 2.21-11.15], and the OR for those with fingernail and toenail involvement to have NDO was 2.66 (95% CI: 1.61-4.34). The F/M OR to have NDO was 2.36 (95% CI: 1.51-3.61), and 9.80 for diabetics (95% CI: 1.01-106.85). The OR for patients with paronychia to have NDO was 10.33 (95% CI: 5.61-18.88) compared to those without paronychia. Compared to patients with a non-wet occupation, the OR for those with a wet occupation to have NDO was 4.76 (95% CI: 2.01-11.16). The distribution of pathogens significantly varies with the involved sites, patients' gender and occupation, and presence of diabetes mellitus or paronychia. In contrast to temperate western countries, NDO is more prevalent in the tropics and subtropics including southern Taiwan. PMID- 16262879 TI - Isolation of Cryptococcus species including C. neoformans from cloaca of pigeons. AB - Upper digestive tract of the pigeon (Columba livia) is well known as a reservoir for different species of Cryptococcus, but lower portions are not so frequently studied. In the present study, we tested on selective media a total of 331 pigeon cloacal swabs; Cryptococcus spp. were recovered from 26 (7.85%). Cryptococcus uniguttulatus was isolated from 11 samples (3.32%), C. laurentii from six (1.81%), C. neoformans var. neoformans from six (1.81%) and C. albidus from three of them (0.91%). The results show the importance of pigeon in the cryptococcosis epidemiology as reservoir and carrier for C. neoformans var. neoformans, but also for other Cryptococcus species of increasing clinical interest. PMID- 16262880 TI - Superficial mycoses due to Trichophyton violaceum in Athens, Greece: a 15-year retrospective study. AB - This is a retrospective study on the superficial mycoses due to Trichophyton violaceum in the greater Athens area for the last 15 years - 1989-2003. During this period 104 patients were found to have an infection due to T. violaceum- an incidence rate of 0.75% of all dermatophytosis. Of the patients 59 were Greeks, 15 Greek Gypsies and 30 immigrants mostly from Albania (50%). Of them 58 were children, 46 adults (mainly women, 34 cases). Trichophyton violaceum infection was presented with a variety of manifestations (127 cases). The prevailing was tinea capitis present in 85 patients - 57 children, 24 women and four men (women : men 6 : 1). Tinea capitis together with other forms of the infection was found in 14 patients. Tinea facie, corporis, manuum, barbae and unguium were seen in nineteen patients. The isolation rate of T. violaceum infection in the Greek population remained at a low level for three decades after the mid-1960s. However, a substantial increase in the isolation rate is observed in the mid 1990s attributed mainly to the influx of economic immigrants from countries where the infection is endemic. PMID- 16262881 TI - Onychomycosis in children: a survey of 46 cases. AB - This is a retrospective study of the agents, clinical aspects, sources of infection and therapy of onychomycosis in children. In the period 1989-2000, we observed 46 consecutive children, until 16 years of age with onychomycosis (29 boys, 17 girls, mean age 10.8 years). Dermatophytes were isolated in 30 cases (Trichophyton rubrum in 22 cases, Trichophyton mentagrophytes in five, Epidermophyton floccosum in two and Trichophyton violaceum in one) and Candida spp. in 16, associated with Trichophyton rubrum in two. Moulds were isolated in three children (Fusarium oxysporum in one, Scopulariopsis brevicaulis in another and Aspergillus fumigatus associated with Trichophyton rubrum in a third). The commonest features were distal and distolateral subungual hyperkeratosis in dermatophyte infections (93%) and onychodystrophy and paronychia in Candida infections (56% and 50% respectively). Forty patients achieved clinical and mycological recovery. It is appropriate to suspect onychomycosis in children, perform microbiological diagnosis and undertake early treatment. An approach of this kind may help to prevent nail dystrophy and the spread of infection. PMID- 16262882 TI - Accurate identification of amphotericin B liposome for injection (AmBisome). PMID- 16262883 TI - A case of pityriasis versicolor atrophicans. AB - A case of pityriasis versicolor atrophicans caused by Malassezia globosa is reported in a 49-year-old woman. Diagnosis was based on direct microscopic examination and culture. Differential diagnosis was performed with respect to other skin diseases manifesting with cutaneous atrophy. PMID- 16262884 TI - Role of Chrysosporium keratinophillum in the parasitic evolution of dermatophytes. AB - Anti-dermatophytic activity of Chrysosporium keratinophillum against species of the genera Trichophyton, Microsporum and Epidermophyton floccosum was tested in vitro. When C. keratinophillum and different species of dermatophytes were inoculated on Sabouraud's dextrose agar plates 2 cm apart, no antagonistic effect of C. keratinophillum on the mycelial growth of dermatophytes was observed. However, conidia production was not observed on the hyphae of Trichophyton rubrum, Trichophyton tonsurans and E. floccosum grown near C. keratinophillum. The secretory substances released by C. keratinophillum inhibited the growth of T. rubrum, T. tonsurans, Trichophyton mentagrophytes var. interdigitale and E. floccosum at a concentration of 2,000 microg ml(-1) when tested by broth dilution technique. No inhibition of the growth was observed for Microsporum gypseum and Microsporum nanum. The anti-fungal activity of secretory substances released by C. keratinophillum was recorded to be heat stable. Results of the present study suggest that the anti-dermatophytic activity of the secretory substances of C. keratinophillum on T. rubrum, T. mentagrophytes var. interdigitale, T. tonsurans and E. floccosum may be responsible in part, for the absence of these dermatophyte species in soil. Considering the global prevalence of C. keratinophillum in soil one may speculate that the anti-dermatophytic activity of C. keratinophillum is one of the early events for the evolutionary divergence of saprophytic archi-dermatophytes to obligate parasitic dermatophyte species. PMID- 16262885 TI - Posaconazole therapy for systemic coccidioidomycosis in a chimpanzee (Pan troglodytes): a case report. AB - Systemic coccidioidomycosis was diagnosed in a 4-year-old male chimpanzee (Pan troglodytes) with ascites and failure to thrive. Physical examination, laboratory and radiological studies, and exploratory laparotomy showed signs of systemic fungal infection that included penetration into the central nervous system (CNS). Serum and cerebrospinal fluid (CSF) titres, along with ascites cytology findings, confirmed the presence of Coccidioides immitis. However, the organism could not be cultured from the CSF. Treatment with fluconazole 10 mg kg(-1) daily for 6 months was not associated with clinical improvement. Subsequent treatment with posaconazole 50 mg kg(-1) daily for approximately 24 months resulted in negative serum titres and improved clinical status. Illness not directly related to the C. immitis infection caused the chimpanzee's deterioration and eventual killing. Histological examination performed during necropsy provided no evidence of coccidioidomycosis; however, a positive C. immitis serum antibody titre was noted. The successful suppression of coccidioidomycosis observed in this chimpanzee suggests that posaconazole may have a role in the treatment of CNS infections caused by susceptible fungi. PMID- 16262886 TI - Bipolaris keratomycosis. AB - A case of mycotic keratitis caused by Bipolaris spicifera is reported. The corneal ulcer developed after 1 week of treatment with corticosteroids and antibiotics. Septate, branched, dematiaceous hyphal elements were detected in the corneal scrapings on two occasions and repeated cultures were positive for this fungus. The infection was resolved with natamycin and itraconazole although the corneal opacity remained and the patient is now waiting for a keratoplasty. PMID- 16262887 TI - Invasive chromoblastomycosis and sinusitis due to Phialophora verrucosa in a child from northern Africa. AB - We report on a severe, verrucous facial mycosis and sinusitis in a 12-year-old Libyan girl. Her disease started with verrucous, hyperkeratotic plaques and subcutaneous violet nodules of unknown origin on her face and upper extremities. Despite topical antimycotic therapy she needed in-hospital treatment because of severely progressive tumorous cutaneous and nasal lesions. Microscopic examination of scale samples taken from the upper extremities and the face revealed brown, thick-walled fungal elements. Under the assumption of a chromoblastomycosis, an oral treatment with itraconazole and fluorocytosin was initiated, with significant improvement of the lesions. The aetiological agent was identified as Phialophora verrucosa. PMID- 16262888 TI - Surface pretreatment for prolonged survival of cemented tibial prosthesis components: full- vs. surface-cementation technique. AB - BACKGROUND: One of few persisting problems of cemented total knee arthroplasty (TKA) is aseptic loosening of tibial component due to degradation of the interface between bone cement and metallic tibial shaft component, particularly for surface cemented tibial components. Surface cementation technique has important clinical meaning in case of revision and for avoidance of stress shielding. Degradation of the interface between bone cement and bone may be a secondary effect due to excessive crack formation in bone cement starting at the opposite metallic surface. METHODS: This study was done to prove crack formation in the bone cement near the metallic surface when this is not coated. We propose a newly developed coating process by PVD layering with SiOx to avoid that crack formation in the bone cement. A biomechanical model for vibration fatigue test was done to simulate the physiological and biomechanical conditions of the human knee joint and to prove excessive crack formation. RESULTS: It was found that coated tibial components showed a highly significant reduction of cement cracking near the interface metal/bone cement (p < 0.01) and a significant reduction of gap formation in the interface metal-to-bone cement (p < 0.05). CONCLUSION: Coating dramatically reduces hydrolytic- and stress-related crack formation at the prosthesis interface metal/bone cement. This leads to a more homogenous load transfer into the cement mantle which should reduce the frequency of loosening in the interfaces metal/bone cement/bone. With surface coating of the tibial component it should become possible that surface cemented TKAs reveal similar loosening rates as TKAs both surface and stem cemented. This would be an important clinical advantage since it is believed that surface cementing reduces metaphyseal bone loss in case of revision and stress shielding for better bone health. PMID- 16262889 TI - 23 years of the discovery of Helicobacter pylori: is the debate over? AB - The Gram negative curved bacillus H. pylori has become the prize bug of all times. Barry Marshall and Robin Warren the two discoverers of this organism have been awarded with this year's Nobel Prize. The Nobel committee at the Karolinska Institute of Sweden has selected this paradigm shift discovery of 1982 as the most impacting in medical sciences. This award has surprised many as the Nobel assembly has selected this 'Robert Koch styled medical detective work' for the prize as compared to many outstanding basic research stories on the waitlist. This editorial briefly touches the significant impact of H. pylori on gastroduodenal management and the path forward as the bug has become quite controversial in recent times. PMID- 16262890 TI - Family history and stroke outcome in a bi-ethnic, population-based stroke surveillance study. AB - BACKGROUND: The genetic epidemiology of ischemic stroke remains relatively unstudied, and information about the genetic epidemiology of ischemic stroke in populations with significant minority representation is currently unavailable. METHODS: The Brain Attack Surveillance in Corpus Christi project (BASIC) is a population-based stroke surveillance study conducted in the bi-ethnic community of Nueces County, Texas, USA. Completed ischemic strokes were identified among patients 45 years or older seen at hospitals in the county between January 1, 2000-December 31, 2002. A random sample of ischemic stroke patients underwent an in-person interview and detailed medical record abstraction (n = 400). Outcomes, including initial stroke severity (NIH stroke scale), age at stroke onset, 90-day mortality and functional outcome (modified Rankin scale > or = 2), were studied for their association with family history of stroke among a first degree relative using multivariable logistic and linear regression. A chi-square test was used to test the association between family history of stroke and ischemic stroke subtype. RESULTS: The study population was 53.0% Mexican American and 58.4% female. Median age was 73.2 years. Forty percent reported a family history of stroke among a first degree relative. Family history of stroke was borderline significantly associated with stroke subtype (p = 0.0563). Family history was associated with poor functional outcome in the multivariable model (OR = 1.87; 95% CI: 1.14-3.09). Family history was not significantly related to initial stroke severity, age at stroke onset, or 90-day mortality. CONCLUSION: Family history of stroke was related to ischemic stroke subtype and to functional status at discharge. More research is needed to understand whether stroke subtype would be a useful selection criterion for genetic association studies and to hypothesize about a possible genetic link to recovery following ischemic stroke. PMID- 16262891 TI - Aging syndrome genes and premature coronary artery disease. AB - BACKGROUND: Vascular disease is a feature of aging, and coronary vascular events are a major source of morbidity and mortality in rare premature aging syndromes. One such syndrome is caused by mutations in the lamin A/C (LMNA) gene, which also has been implicated in familial insulin resistance. A second gene related to premature aging in man and in murine models is the KLOTHO gene, a hypomorphic variant of which (KL-VS) is significantly more common in the first-degree relatives of patients with premature coronary artery disease (CAD). We evaluated whether common variants at the LMNA or KLOTHO genes are associated with rigorously defined premature CAD. METHODS: We identified 295 patients presenting with premature acute coronary syndromes confirmed by angiography. A control group of 145 patients with no evidence of CAD was recruited from outpatient referral clinics. Comprehensive haplotyping of the entire LMNA gene, including the promoter and untranslated regions, was performed using a combination of TaqMan probes and direct sequencing of 14 haplotype-tagging single nucleotide polymorphisms (SNPs). The KL-VS variant of the KLOTHO gene was typed using restriction digest of a PCR amplicon. RESULTS: Two SNPs that were not in Hardy Weinberg equilibrium were excluded from analysis. We observed no significant differences in allele, genotype or haplotype frequencies at the LMNA or KLOTHO loci between the two groups. In addition, there was no evidence of excess homozygosity at the LMNA locus. CONCLUSION: Our data do not support the hypothesis that premature CAD is associated with common variants in the progeroid syndrome genes LMNA and KLOTHO. PMID- 16262893 TI - Opportunities for using spatial property assessment data in air pollution exposure assessments. AB - BACKGROUND: Many epidemiological studies examining the relationships between adverse health outcomes and exposure to air pollutants use ambient air pollution measurements as a proxy for personal exposure levels. When pollution levels vary at neighbourhood levels, using ambient pollution data from sparsely located fixed monitors may inadequately capture the spatial variation in ambient pollution. A major constraint to moving toward exposure assessments and epidemiological studies of air pollution at a neighbourhood level is the lack of readily available data at appropriate spatial resolutions. Spatial property assessment data are widely available in North America and may provide an opportunity for developing neighbourhood level air pollution exposure assessments. RESULTS: This paper provides a detailed description of spatial property assessment data available in the Pacific Northwest of Canada and the United States, and provides examples of potential applications of spatial property assessment data for improving air pollution exposure assessment at the neighbourhood scale, including: (1) creating variables for use in land use regression modelling of neighbourhood levels of ambient air pollution; (2) enhancing wood smoke exposure estimates by mapping fireplace locations; and (3) using data available on individual building characteristics to produce a regional air pollution infiltration model. CONCLUSION: Spatial property assessment data are an extremely detailed data source at a fine spatial resolution, and therefore a source of information that could improve the quality and spatial resolution of current air pollution exposure assessments. PMID- 16262892 TI - Asbestosis in an asbestos composite mill at Mumbai: a prevalence study. AB - BACKGROUND: Of an estimated 100,000 workers exposed to asbestos in India, less than 30 have been compensated. The reasons for such a small number are: refusal by management sponsored studies to grant medical certifications to workers suffering from occupational diseases, lack of training for doctors in diagnosis of occupational lung diseases, deliberate misdiagnosis by doctors of asbestosis as either chronic bronchitis or tuberculosis and the inherent class bias of middle class doctors against workers. The aim of the study was to identify workers suffering from Asbestosis (parenchymal and pleural non-malignant disease) among the permanent workers of the Hindustan Composites Factory and assess their disability and medically certify them, whereupon they could avail of their basic rights to obtain compensation and proper treatment. METHODS: The study was conducted by the Occupational Health and Safety Centre and the Workers' Union. Asbestosis was diagnosed if they had an occupational history of asbestos exposure for at least 15 years and showed typical radiographic findings. RESULTS: Of 232 workers in the factory, 181 participated in the survey. 22% of them had asbestosis. All the asbestos affected workers had at least 20 years of exposure. 7% had rhonchi, 34% had late basal inspiratory rates, 82% had more than 80% of Forced Expiratory Volume in the first second (FEV1)/Forced Vital capacity (FVC) ratio and 66% had FVC less than 80% of the predicted value. On radiology 7% had only pleural disease, 10% had both pleural and parenchymal disease and 82% had only parenchymal disease. The association of pleural disease with chest pain was statistically significant. CONCLUSION: We found the prevalence of asbestosis among exposed workers to be less than that anticipated for the number of years of exposure due to "Healthy Worker Effect". We suggest that all affected asbestos workers (including those who have been forced to leave) in India be medically certified and compensated. We also recommend better control of asbestos use in India. We also implore the management to provide all information about the work process and its hazards, conduct medical checkups as mandated by law and give the medical records to the workers. PMID- 16262894 TI - Viral load responses to HAART is an independent predictor of a new AIDS event in late stage HIV infected patients: prospective cohort study. AB - BACKGROUND: A sizeable number of HIV-infected patients receiving HAART do not maintain prolonged virologic suppression. We evaluated long-term HIV viral load (VL) responses to HAART as a risk factor for AIDS events (AE) that is independent of CD4 responses. METHODS: A cohort of patients with pre-therapy CD4 < 200/mm3 who had CD4 and VL measurements for > one year after receiving HAART at a university clinic were prospectively enrolled. Cox proportional multivariate regression model was used to determine whether CD4 and VL responses were independently associated with new AE. RESULTS: The patient (N = 214) mean baseline CD4 = 92/mm3, VL = 219,000 c/mL and follow-up duration 42.3 months (range 13-72 months). A new AE occurred in 56 patients; CD4 cell count response to HAART that remained < 200/mm3 throughout the study period was a significant risk factor for new AE (RR = 9.7-12.5; p < 0.001). Similarly, VL responses that remained > 5,000 c/mL during this period was also a significant risk factor (RR = 6.7-12.8; p = 0.001) that was independent of CD4 response adjusted for <> 200/mm3. CONCLUSION: Maintaining adequate long-term virologic responses to HAART provides a clinical benefit independent of CD4 responses. PMID- 16262895 TI - SNiPer: improved SNP genotype calling for Affymetrix 10K GeneChip microarray data. AB - BACKGROUND: High throughput microarray-based single nucleotide polymorphism (SNP) genotyping has revolutionized the way genome-wide linkage scans and association analyses are performed. One of the key features of the array-based GeneChip Mapping 10K Array from Affymetrix is the automated SNP calling algorithm. The Affymetrix algorithm was trained on a database of ethnically diverse DNA samples to create SNP call zones that are used as static models to make genotype calls for experimental data. We describe here the implementation of clustering algorithms on large training datasets resulting in improved SNP call rates on the 10K GeneChip. RESULTS: A database of 948 individuals genotyped on the GeneChip Mapping 10K 2.0 Array was used to identify 822 SNPs that were called consistently less than 75% of the time. These SNPs represent on average 8.25% of the total SNPs on each chromosome with chromosome 19, the most gene-rich chromosome, containing the highest proportion of poor performers (18.7%). To remedy this, we created SNiPer, a new application which uses two clustering algorithms to yield increased call rates and equivalent concordance to Affymetrix called genotypes. We include a training set for these algorithms based on individual genotypes for 705 samples. SNiPer has the capability to be retrained for lab-specific training sets. SNiPer is freely available for download at http://www.tgen.org/neurogenomics/data. CONCLUSION: The correct calling of poor performing SNPs may prove to be key in future linkage studies performed on the 10K GeneChip. It would prove particularly invaluable for those diseases that map to chromosome 19, known to contain a high proportion of poorly performing SNPs. Our results illustrate that SNiPer can be used to increase call rates on the 10K GeneChip without sacrificing accuracy, thereby increasing the amount of valid data generated. PMID- 16262897 TI - Increased oxidative stress associated with the severity of the liver disease in various forms of hepatitis B virus infection. AB - BACKGROUND: Oxidative stress can be defined as an increase in oxidants and/or a decrease in antioxidant capacity. There is limited information about the oxidative status in subjects with hepatitis B virus infection. We aimed to evaluate the oxidative status in patients with various clinical forms of chronic hepatitis B infection. METHODS: Seventy-six patients with hepatitis B virus infection, in whom 33 with chronic hepatitis, 31 inactive carriers and 12 with cirrhosis, and 16 healthy subjects were enrolled. Total antioxidant response and total peroxide level measurement, and calculation of oxidative stress index were performed in all participants. RESULTS: Total antioxidant response was significantly lower in cirrhotics than inactive HbsAg carriers and controls (p = 0.008 and p = 0.008, respectively). Total peroxide level and oxidative stress index was significantly higher in cirrhotic (p < 0.001, both) and chronic hepatitis B subjects (p < 0.001, both) than inactive HbsAg carriers and controls. Total antioxidant response was comparable in chronic hepatitis B subjects, inactive HbsAg carriers and controls (both, p > 0.05/6). Total peroxide level and oxidative stress index were also comparable in inactive HBsAg carriers and controls (both, p > 0.05/6). Serum alanine amino transferase level was positively correlated with total peroxide level and oxidative stress index only in chronic hepatitis B subjects (p = 0.002, r = 0.519 and p = 0.008, r = 0.453, respectively). CONCLUSION: Oxidative stress occurs secondarily to increased total lipid peroxidation and inadequate total antioxidant response and is related to severity of the disease and replication status of virus in hepatitis B infection. PMID- 16262898 TI - Back pain reporting in young girls appears to be puberty-related. AB - BACKGROUND: There is a large increase in back pain reporting in the early teens. In no previous study has the prevalence of low back pain been investigated in relation to the onset of puberty. The objective of this study was to establish whether the onset of puberty is associated with back pain reporting in young girls. METHODS: A subsample of 254 girls aged 8-10 years and 165 girls aged 14-16 years from a cross-sectional survey of 481 children aged 8-10 years and 325 adolescents aged 14-16 years of both sexes. Main outcome measures were back pain defined as low back pain, mid back pain, and/or neck pain in the past month. Other variables of interest were Puberty (five different stages), age, body mass index, and smoking. Independent information on onset of puberty was obtained through a physical examination and on back pain through an individual structured interview. The association was studied between onset of puberty and the outcome variable (the one month period prevalence of back pain), controlling for overweight, and smoking. Odds ratios with 95% confidence intervals were used to describe bivariate associations, logistic regression with robust standard errors was used for multivariate analyses. RESULTS: There is a highly significant trend for increased back pain reporting with increasing level of puberty until maturity is reached. The biggest leap appears between the second level (beginning of puberty) and the third level (mid puberty) and the findings remain after controlling for the covariates. These results emanate from the low back, whereas pain in the mid back and neck do not seem to be linked with pubertal stage. CONCLUSION: In girls, the reporting of low back pain increases in frequency during puberty until maturity, regardless of age. Why some girls are susceptible to back pain in the early stage of puberty is unknown. PMID- 16262896 TI - The response of VEGF-stimulated endothelial cells to angiostatic molecules is substrate-dependent. AB - BACKGROUND: The microenvironment surrounding cells can exert multiple effects on their biological responses. In particular the extracellular matrix surrounding cells can profoundly influence their behavior. It has been shown that the extracellular matrix composition in tumors is vastly different than that found in normal tissue with increased amounts of certain matrices such as collagen I. It has been previously demonstrated that VEGF stimulation of endothelial cells growing on type I collagen results in the induction of bcl-2 expression and enhanced endothelial cell survival. We sought to investigate whether this increased endothelial cell survival resulted in the failure of angiostatic molecules to inhibit angiogenesis. RESULTS: We now demonstrate that VEGF-induced survival on collagen I impairs the ability of three known angiostatic molecules, TSP-1, IP-10 and endostatin to inhibit endothelial cell proliferation. Apoptosis of endothelial cells, growing on collagen I, induced by TSP-1 and IP-10 was also inhibited following VEGF stimulation. In contrast, endostatin induced apoptosis in these same cells. Further analysis determined that endostatin did not decrease the expression of bcl-2 nor did it increase activation of caspase-3 in the presence of VEGF. Alternatively, it appeared that in the presence of VEGF, endostatin induced the activation of caspase-8 in endothelial cells grown on collagen I. Furthermore, only endostatin had the ability to inhibit VEGF-induced sprout formation in collagen I gels. CONCLUSION: These data suggest that TSP-1, IP-10 and endostatin inhibit endothelial cells via different mechanisms and that only endostatin is effective in inhibiting angiogenic activities in the presence of collagen I. Our results suggest that the efficacy of angiostatic treatments may be impaired depending on the context of the extracellular matrix within the tumor environment and thus could impede the efficacy of angiostatic therapies. PMID- 16262901 TI - "Near-miss" obstetric events and maternal deaths in Sagamu, Nigeria: a retrospective study. AB - AIM: To determine the frequency of near-miss (severe acute maternal morbidity) and the nature of near-miss events, and comparatively analysed near-miss morbidities and maternal deaths among pregnant women managed over a 3-year period in a Nigerian tertiary centre. METHODS: Retrospective facility-based review of cases of near-miss and maternal death which occurred between 1 January 2002 and 31 December 2004. Near-miss case definition was based on validated disease specific criteria, comprising of five diagnostic categories: haemorrhage, hypertensive disorders in pregnancy, dystocia, infection and anaemia. The near miss morbidities were compared with maternal deaths with respect to demographic features and disease profiles. Mortality indices were determined for various disease processes to appreciate the standard of care provided for life threatening obstetric conditions. The maternal death to near-miss ratios for the three years were compared to assess the trend in the quality of obstetric care. RESULTS: There were 1501 deliveries, 211 near-miss cases and 44 maternal deaths. The total near-miss events were 242 with a decreasing trend from 2002 to 2004. Demographic features of cases of near-miss and maternal death were comparable. Besides infectious morbidity, the categories of complications responsible for near-misses and maternal deaths followed the same order of decreasing frequency. Hypertensive disorders in pregnancy and haemorrhage were responsible for 61.1% of near-miss cases and 50.0% of maternal deaths. More women died after developing severe morbidity due to uterine rupture and infection, with mortality indices of 37.5% and 28.6%, respectively. Early pregnancy complications and antepartum haemorrhage had the lowest mortality indices. Majority of the cases of near-miss (82.5%) and maternal death (88.6%) were unbooked for antenatal care and delivery in this hospital. Maternal mortality ratio for the period was 2931.4 per 100,000 deliveries. The overall maternal death to near-miss ratio was 1: 4.8 and this remained relatively constant over the 3-year period. CONCLUSION: The quality of care received by critically ill obstetric patients in this centre is suboptimal with no evident changes between 2002 and 2004. Reduction of the present maternal mortality ratio may best be achieved by developing evidence-based protocols and improving the resources for managing severe morbidities due to hypertension and haemorrhage especially in critically ill unbooked patients. Tertiary care hospitals in Nigeria could also benefit from evaluation of their standard of obstetric care by including near-miss investigations in their maternal death enquiries. PMID- 16262899 TI - The bone marrow aspirate and biopsy in the diagnosis of unsuspected nonhematologic malignancy: a clinical study of 19 cases. AB - BACKGROUND: Although bone marrow metastases can be found commonly in some malignant tumors, diagnosing a nonhematologic malignancy from marrow is not a usual event. METHODS: To underscore the value of bone marrow aspiration and biopsy as a short cut in establishing a diagnosis for disseminated tumors, we reviewed 19 patients with nonhematologic malignancies who initially had diagnosis from bone marrow. RESULTS: The main indications for bone marrow examination were microangiopathic hemolytic anemia (MAHA), leukoerythroblastosis (LEB) and unexplained cytopenias. Bone marrow aspiration was not diagnostic due to dry tap or inadequate material in 6 cases. Biopsy results were parallel to the cytological ones in all cases except one; however a meticulous second examination of the biopsy confirmed the cytologic diagnosis in this patient too. The most common histologic subtype was adenocarcinoma, and after all the clinical and laboratory evaluations, the primary focus was disclosed definitively in ten patients (5 stomach, 3 prostate, 1 lung, 1 muscle) and probably in four patients (3 gastrointestinal tract, 1 lung). All work up failed in five patients and these cases were classified as tumor of unknown origin (TUO). CONCLUSION: Our series showed that anemia, thrombocytopenia, elevated red cell distribution width (RDW) and hypoproteinemia formed a uniform tetrad in patients with disseminated tumors that were diagnosed via bone marrow examination. The prognosis of patients was very poor and survivals were only a few days or weeks (except for 4 patients whose survivals were longer). We concluded that MAHA, LEB and unexplained cytopenias are strong indicators of the necessity of bone marrow examination. Because of the very short survival of many patients, all investigational procedures should be judged in view of their rationality, and should be focused on treatable primary tumors. PMID- 16262900 TI - Inhaled tolafentrine reverses pulmonary vascular remodeling via inhibition of smooth muscle cell migration. AB - BACKGROUND: The aim of the study was to assess the chronic effects of combined phosphodiesterase 3/4 inhibitor tolafentrine, administered by inhalation, during monocrotaline-induced pulmonary arterial hypertension (PAH) in rats. METHODS: CD rats were given a single subcutaneous injection of monocrotaline to induce PAH. Four weeks after, rats were subjected to inhalation of tolafentrine or sham nebulization in an unrestrained, whole body aerosol exposure system. In these animals (i) the acute pulmonary vasodilatory efficacy of inhaled tolafentrine (ii) the anti-remodeling effect of long-term inhalation of tolafentrine (iii) the effects of tolafentrine on the expression profile of 96 genes encoding cell adhesion and extracellular matrix regulation were examined. In addition, the inhibitory effect of tolafentrine on ex vivo isolated pulmonary artery SMC cell migration was also investigated. RESULTS: Monocrotaline injection provoked severe PAH (right ventricular systolic pressure increased from 25.9 +/- 4.0 to 68.9 +/- 3.2 after 4 weeks and 74.9 +/- 5.1 mmHg after 6 weeks), cardiac output depression and right heart hypertrophy. The media thickness of the pulmonary arteries and the proportion of muscularization of small precapillary resistance vessels increased dramatically, and the migratory response of ex-vivo isolated pulmonary artery smooth muscle cells (PASMC) was increased. Micro-arrays and subsequent confirmation with real time PCR demonstrated upregulation of several extracellular matrix regulation and adhesion genes, such as matrixmetalloproteases (MMP) 2, 8, 9, 10, 11, 12, 20, Icam, Itgax, Plat and serpinb2. When chronically nebulized from day 28 to 42 (12 daily aerosol maneuvers), after full establishment of severe pulmonary hypertension, tolafentrine reversed about 60% of all hemodynamic abnormalities, right heart hypertrophy and monocrotaline-induced structural lung vascular changes, including the proportion of pulmonary artery muscularization. The upregulation of extracellular matrix regulation and adhesion genes was reduced by nearly 80% by inhalation of the tolafentrine. When assessed in vitro, tolafentrine blocked the enhanced PASMC migratory response. CONCLUSION: In conclusion, we demonstrate for the first time that inhalation of combined PDE3/4 inhibitor reverses pulmonary hypertension fully developed in response to monocrotaline in rats. This "reverse remodeling" effect includes structural changes in the lung vascular wall and key molecular pathways of matrix regulation, concomitant with 60% normalization of hemodynamics. PMID- 16262902 TI - Translating research into policy and practice in developing countries: a case study of magnesium sulphate for pre-eclampsia. AB - BACKGROUND: The evidence base for improving reproductive health continues to grow. However, concerns remain that the translation of this evidence into appropriate policies is partial and slow. Little is known about the factors affecting the use of evidence by policy makers and clinicians, particularly in developing countries. The objective of this study was to examine the factors that might affect the translation of randomised controlled trial (RCT) findings into policies and practice in developing countries. METHODS: The recent publication of an important RCT on the use of magnesium sulphate to treat pre-eclampsia provided an opportunity to explore how research findings might be translated into policy. A range of research methods, including a survey, group interview and observations with RCT collaborators and a survey of WHO drug information officers, regulatory officials and obstetricians in 12 countries, were undertaken to identify barriers and facilitators to knowledge translation. RESULTS: It proved difficult to obtain reliable data regarding the availability and use of commonly used drugs in many countries. The perceived barriers to implementing RCT findings regarding the use of magnesium sulphate for pre-eclampsia include drug licensing and availability; inadequate and poorly implemented clinical guidelines; and lack of political support for policy change. However, there were significant regional and national differences in the importance of specific barriers. CONCLUSION: The policy changes needed to ensure widespread availability and use of magnesium sulphate are variable and complex. Difficulties in obtaining information on availability and use are combined with the wide range of barriers across settings, including a lack of support from policy makers. This makes it difficult to envisage any single intervention strategy that might be used to promote the uptake of research findings on magnesium sulphate into policy across the study settings. The publication of important trials may therefore not have the impacts on health care that researchers hope for. PMID- 16262903 TI - Testing assumptions for endophenotype studies in ADHD: reliability and validity of tasks in a general population sample. AB - BACKGROUND: Advances in both genetic and cognitive-experimental studies on attention deficit hyperactivity disorder (ADHD) have opened new opportunities for cognitive endophenotype research. In such genetic designs the focus is on individual differences in characteristics, associated with ADHD, that can be measured reliably over time. Genetic studies that take a 'quantitative trait loci' approach hypothesise that multiple susceptibility genes contribute to a continuous dimension of ADHD symptoms. As an important initial step, we aimed to investigate the underlying assumptions that (1) key cognitive-experimental tasks indicate adequate test-retest reliability and (2) ADHD symptom scores in a general population sample are associated with performance on these tasks. METHODS: Forty-nine children were assessed on a go/no-go task and a reaction time task (the 'fast task') that included manipulations with event rate and incentives. The children were assessed twice, with a test-retest interval of two weeks. RESULTS: The majority of the task variables demonstrated moderate-to-good test-retest reliability. The correlations between teacher ratings of ADHD symptoms and key task variables were .4-.6: ADHD symptoms were associated with poor performance (especially high reaction time variability) in a slow baseline condition, whereas there was low or no association in conditions with a faster event rate or incentives. In contrast, no clear pattern of findings emerged based on parent ratings of ADHD symptoms. CONCLUSION: The data support the usefulness of the go/no-go and fast tasks for genetic studies, which require reliable and valid indices of individual differences. The overall pattern of associations between teacher ratings of ADHD symptoms and task variables is consistent with effects of event rate and incentives on performance, as predicted by the model of activation and arousal regulation. The lack of a clear pattern of findings with parent ratings of ADHD symptoms warrants further study. PMID- 16262904 TI - EDTA chelation therapy for cardiovascular disease: a systematic review. AB - BACKGROUND: Numerous practitioners of both conventional and complementary and alternative medicine throughout North America and Europe claim that chelation therapy with EDTA is an effective means to both control and treat cardiovascular disease. These claims are controversial, and several randomized controlled trials have been completed dealing with this topic. To address this issue we conducted a systematic review to evaluate the best available evidence for the use of EDTA chelation therapy in the treatment of cardiovascular disease. METHODS: We conducted a systematic review of 7 databases from inception to May 2005. Hand searches were conducted in review articles and in any of the trials found. Experts in the field were contacted and registries of clinical trials were searched for unpublished data. To be included in the final systematic review, the studies had to be randomized controlled clinical trials. RESULTS: A total of seven articles were found assessing EDTA chelation for the treatment of cardiovascular disease. Two of these articles were subgroup analyses of one RCT that looked at different clinical outcomes. Of the remaining five studies, two smaller studies found a beneficial effect whereas the other three exhibited no benefit for cardiovascular disease from the use of EDTA chelation therapy. Adverse effects were rare but those of note included a few cases of hypocalcemia and a single case of increased creatinine in a patient on the EDTA intervention. CONCLUSION: The best available evidence does not support the therapeutic use of EDTA chelation therapy in the treatment of cardiovascular disease. Although not considered to be a highly invasive or harmful therapy, it is possible that the use of EDTA chelation therapy in lieu of proven therapy may result in causing indirect harm to the patient. PMID- 16262905 TI - Blood pressure patterns in rural, semi-urban and urban children in the Ashanti region of Ghana, West Africa. AB - BACKGROUND: High blood pressure, once rare, is rapidly becoming a major public health burden in sub-Saharan/Africa. It is unclear whether this is reflected in children. The main purpose of this study was to assess blood pressure patterns among rural, semi-urban, and urban children and to determine the association of blood pressure with locality and body mass index (BMI) in this sub-Saharan Africa setting. METHODS: We conducted a cross-sectional survey among school children aged 8-16 years in the Ashanti region of Ghana (West-Africa). There were 1277 children in the study (616 boys and 661 females). Of these 214 were from rural, 296 from semi-urban and 767 from urban settings. RESULTS: Blood pressure increased with increasing age in rural, semi-urban and urban areas, and in both boys and girls. The rural boys had a lower systolic and diastolic blood pressure than semi-urban boys (104.7/62.3 vs. 109.2/66.5; p < 0.001) and lower systolic blood pressure than urban boys (104.7 vs. 107.6; p < 0.01). Girls had a higher blood pressure than boys (109.1/66.7 vs. 107.5/63.8; p < 0.01). With the exception of a lower diastolic blood pressure amongst rural girls, no differences were found between rural girls (107.4/64.4) and semi-urban girls (108.0/66.1) and urban girls (109.8/67.5). In multiple linear regression analysis, locality and BMI were independently associated with blood pressure in both boys and girls. CONCLUSION: These findings underscore the urgent need for public health measures to prevent increasing blood pressure and its sequelae from becoming another public health burden. More work on blood pressure in children in sub-Saharan African and other developing countries is needed to prevent high blood pressure from becoming a major burden in many of these countries. PMID- 16262906 TI - Processing sites in the human immunodeficiency virus type 1 (HIV-1) Gag-Pro-Pol precursor are cleaved by the viral protease at different rates. AB - We have examined the kinetics of processing of the HIV-1 Gag-Pro-Pol precursor in an in vitro assay with mature protease added in trans. The processing sites were cleaved at different rates to produce distinct intermediates. The initial cleavage occurred at the p2/NC site. Intermediate cleavages occurred at similar rates at the MA/CA and RT/IN sites, and to a lesser extent at sites upstream of RT. Late cleavages occurred at the sites flanking the protease (PR) domain, suggesting sequestering of these sites. We observed paired intermediates indicative of half- cleavage of RT/RH site, suggesting that the RT domain in Gag Pro-Pol was in a dimeric form under these assay conditions. These results clarify our understanding of the processing kinetics of the Gag-Pro-Pol precursor and suggest regulated cleavage. Our results further suggest that early dimerization of the PR and RT domains may serve as a regulatory element to influence the kinetics of processing within the Pol domain. PMID- 16262908 TI - General practice and the New Zealand health reforms--lessons for Australia? AB - New Zealand's health sector has undergone three significant restructures within 10 years. The most recent has involved a Primary Health Care Strategy, launched in 2001. Primary Health Organisations (PHOs), administered by 21 District Health Boards, are the local structures for implementing the Primary Health Care Strategy. Ninety-three percent of the New Zealand population is now enrolled within 79 PHOs, which pose a challenge to the well-established Independent Practitioner Associations (IPAs). Although there was initial widespread support for the philosophy underlying the Primary Health Care Strategy, there are concerns amongst general practitioners (GPs) and their professional organisations relating to its implementation. These centre around 6 main issues: 1. Loss of autonomy, 2. Inadequate management funding and support, 3. Inconsistency and variations in contracting processes, 4. Lack of publicity and advice around enrollment issues, 5. Workforce and workload issues, 6. Financial risks. On the other hand, many GPs are feeling positive regarding the opportunities for PHOs, particularly for being involved in the provision of a wider range of community health services. Australia has much to learn from New Zealand's latest health sector and primary health care reforms. The key lessons concern: The need for a national primary health care strategy, active engagement of general practitioners and their professional organisations, recognition of implementation costs, the need for infrastructural support, including information technology and quality systems, robust management and governance arrangements, issues related to critical mass and population/distance trade offs in service delivery models. PMID- 16262907 TI - A functional profile of gene expression in ARPE-19 cells. AB - BACKGROUND: Retinal pigment epithelium cells play an important role in the pathogenesis of age related macular degeneration. Their morphological, molecular and functional phenotype changes in response to various stresses. Functional profiling of genes can provide useful information about the physiological state of cells and how this state changes in response to disease or treatment. In this study, we have constructed a functional profile of the genes expressed by the ARPE-19 cell line of retinal pigment epithelium. METHODS: Using Affymetrix MAS 5.0 microarray analysis, genes expressed by ARPE-19 cells were identified. Using GeneChip annotations, these genes were classified according to their known functions to generate a functional gene expression profile. RESULTS: We have determined that of approximately 19,044 unique gene sequences represented on the HG-U133A GeneChip, 6,438 were expressed in ARPE-19 cells irrespective of the substrate on which they were grown (plastic, fibronectin, collagen, or Matrigel). Rather than focus our subsequent analysis on the identity or level of expression of each individual gene in this large data set, we examined the number of genes expressed within 130 functional categories. These categories were selected from a library of HG-U133A GeneChip annotations linked to the Affymetrix MAS 5.0 data sets. Using this functional classification scheme, we were able to categorize about 70% of the expressed genes and condense the original data set of over 6,000 data points into a format with 130 data points. The resulting ARPE-19 Functional Gene Expression Profile is displayed as a percentage of ARPE-19-expressed genes. CONCLUSION: The Profile can readily be compared with equivalent microarray data from other appropriate samples in order to highlight cell-specific attributes or treatment-induced changes in gene expression. The usefulness of these analyses is based on the assumption that the numbers of genes expressed within a functional category provide an indicator of the overall level of activity within that particular functional pathway. PMID- 16262909 TI - The changing epidemiology of pediatric aseptic meningitis in Daejeon, Korea from 1987 to 2003. AB - BACKGROUND: Aseptic meningitis is a relatively frequent childhood disease and virologic data suggest that enteroviruses are the commonest etiologic agents. We evaluated the epidemiologic characteristics of aseptic meningitis in Daejeon, South Korea from 1987 to 2003. METHODS: 2201 medical records of children with aseptic meningitis admitted to The Catholic University of Korea, Daejeon St Mary's Hospital were retrospectively analyzed. RESULTS: Outbreaks of aseptic meningitis were observed in 1990, 1993, 1996, 1997, 2001 and 2002. The age distribution of cases was relatively uniform, with a higher incidence in those aged < 1 year and 4-7 years. The male-to-female ratio was 2:1. There was a higher incidence of disease in the summer (May to August, 74.1% of total). Comparison of the largest epidemics in 1997 and 2002 showed significant differences in the incidence in those < 1 year (11.8% vs. 4.4%, respectively; P = 0.001). Neurologic sequelae were observed in 0.7% of the patients. CONCLUSION: Aseptic meningitis, rare before the 1980s in Korea, has since become a common clinical entity. Since 1990, outbreaks of aseptic meningitis have occurred every 1 to 3 years in Daejeon in keeping with Korea-wide epidemics. The frequency of disease affecting children less than one year of age may reflect herd immunity to the epidemic strain. PMID- 16262910 TI - Pharmacovigilance program to monitor adverse reactions of recombinant streptokinase in acute myocardial infarction. AB - BACKGROUND: Streptokinase (SK) is an effective fibrinolytic agent for the treatment of acute myocardial infarction (AMI). The objective of the present study was to assess the adverse drug reactions (ADRs) associated with intravenous recombinant SK in patients with AMI in routine clinical practice. METHODS: A national, prospective and spontaneous reporting-based pharmacovigilance program was conducted in Cuba. Patient demographics, suspected ADR description, elements to define causality, and outcomes were documented and analyzed. RESULTS: A total of 1496 suspected ADRs identified in 792 patients out of the 1660 (47.7 %) prescriptions reported in the program, were received from July 1995 to July 2002. Most of the patients (71.3%) were male, 67.2% were white and mean age was 61.6 +/ 13.0 years. The mean time interval between the onset of symptoms and the start of the SK infusion was 4.9 +/- 3.7 h. The most frequently reported ADRs were hypotension, arrhythmias, chills, tremors, vomiting, nauseas, allergy, bleeding and fever. ADR severity was 38% mild, 38% moderate, 10% severe, and 4% very severe. Only 3 patients with hemorrhagic stroke were reported. Seventy-two patients died in-hospital mainly because of cardiac causes associated with the patient's underlying clinical condition. Mortality was 3 times more likely in patients suffering arrhythmias than in those without this event (odds ratio 3.1, 95% CI: 1.8 to 5.1). Most of the reported ADRs were classified as possibly or probably associated with the study medication. CONCLUSION: Recombinant SK was associated with a similar post-marketing safety profile to those suggested in previous clinical trials. PMID- 16262911 TI - Association between lifestyle factors and plasma adiponectin levels in Japanese men. AB - BACKGROUND: Adiponectin is an adipocyte-specific protein that plays a role in obesity, insulin resistant, lipid metabolism, and anti-inflammation. Hypoadiponectinemia may be associated with a higher risk for type 2 diabetes and cardiovascular disease. Some studies suggest that adiponectin levels are modulated by lifestyle factors, but little is known about the associations between lifestyle factors and plasma adiponectin levels in Japanese people. We therefore investigated the associations between lifestyle factors and plasma adiponectin levels in general Japanese men. METHODS: The subjects were 202 Japanese male workers who participated in an annual health check. They provided details about anthropometrical data, blood collection, their use of prescribed medication, and the clinical history of their families. They also completed a self-administered questionnaire about their lifestyles. RESULTS: Subjects with plasma adiponectin levels below 4.0 microg/ml had significantly lower levels of HDL cholesterol and higher levels of BMI, SBP, DBP, total cholesterol, FBG, and platelets than did subjects with higher adiponectin levels. In multiple logistic regression after multiple adjustment, a plasma adiponectin level below 4.0 microg/ml was significantly associated with smoking (odds ratio [OR] = 2.08, 95% confidence interval [CI] = 1.01-4.30), a daily diet rich in deep-yellow vegetables (OR = 0.25, 95% CI= 0.07-0.91), frequent eating out (OR = 2.45, 95% CI = 1.19-5.08), and physical exercise two or more times a week (OR = 0.21, 95% CI = 0.06-0.74). CONCLUSION: Our findings show that adiponectin levels in general Japanese men are independently related to smoking, dietary factors, and physical exercise. We think that lifestyle habits might independently modulate adiponectin levels and that adiponectin might be the useful biomarker helping people to avoid developing type 2 diabetes and cardiovascular disease by modifying their lifestyles. PMID- 16262912 TI - Indications and outcome of repeat penetrating keratoplasty in India. AB - BACKGROUND: Repeat penetrating keratoplasty is quite often required as there is high chance of failure of the primary graft particularly in the developing world. We planned a study to analyze the indications and outcome of repeat penetrating keratoplasty in a tertiary care centre in India. METHODS: A retrospective analysis of all the patients who underwent repeat penetrating keratoplasty, between January 1999 and December 2001 was performed. The parameters evaluated were indication for the primary penetrating keratoplasty, causes of failure of the previous graft, and final visual outcome and clarity of the repeat corneal grafts. RESULTS: Of fifty-three eyes of 50 patients with repeat penetrating keratoplasty (three patients underwent bilateral corneal regrafts), 37 eyes had undergone one regraft each, 14 eyes two regrafts and two eyes had three regrafts. The follow-up of the patients ranged from one to three years. The most common primary etiologic diagnosis was vascularized corneal scars (66%), of which the scars related to infection were most common (68.5%). Twenty-eight regrafts (52.8%) remained clear at a mean follow-up of 1.54 +/- 0.68 years, of which 25 were single regrafts (89.3%). The commonest cause of failure of regraft was infection to the corneal graft (recurrence of herpetic infection in 9 eyes and perforated graft ulcers in 3 eyes). Three (18.6%) of the 16 eyes with multiple corneal regrafts achieved a BCVA of 6/60. Overall, only five eyes (all with single regraft) achieved a BCVA of 6/18 or better at the end of follow-up. CONCLUSION: Graft infection is the leading cause of failure of repeat keratoplasty in this part of the world. Prognosis for visual recovery and graft survival is worse in eyes undergoing multiple regrafts. PMID- 16262913 TI - Precis of Breakdown of Will. AB - Behavioral science has long been puzzled by the experience of temptation, the resulting impulsiveness, and the variably successful control of this impulsiveness. In conventional theories, a governing faculty like the ego evaluates future choices consistently over time, discounting their value for delay exponentially, that is, by a constant rate; impulses arise when this ego is confronted by a conditioned appetite. Breakdown of Will (Ainslie 2001) presents evidence that contradicts this model. Both people and nonhuman animals spontaneously discount the value of expected events in a curve where value is divided approximately by expected delay, a hyperbolic form that is more bowed than the rational, exponential curve. With hyperbolic discounting, options that pay off quickly will be temporarily preferred to richer but slower-paying alternatives, a phenomenon that, over periods from minutes to days, can account for impulsive behaviors, and over periods of fractional seconds can account for involuntary behaviors. Contradictory reward-getting processes can in effect bargain with each other, and stable preferences can be established by the perception of recurrent choices as test cases (precedents) in recurrent intertemporal prisoner's dilemmas. The resulting motivational pattern resembles traditional descriptions of the will, as well as of compulsive phenomena that can now be seen as side-effects of will: over-concern with precedent, intractable but circumscribed failures of self-control, a motivated ("dynamic") unconscious, and an inability to exploit emotional rewards. Hyperbolic curves also suggest a means of reducing classical conditioning to motivated choice, the last necessary step for modeling many involuntary processes like emotion and appetite as reward seeking behaviors; such modeling, in turn, provides a rationale for empathic reward and the "construction" of reality. PMID- 16262930 TI - Understanding and sharing intentions: the origins of cultural cognition. AB - We propose that the crucial difference between human cognition and that of other species is the ability to participate with others in collaborative activities with shared goals and intentions: shared intentionality. Participation in such activities requires not only especially powerful forms of intention reading and cultural learning, but also a unique motivation to share psychological states with others and unique forms of cognitive representation for doing so. The result of participating in these activities is species-unique forms of cultural cognition and evolution, enabling everything from the creation and use of linguistic symbols to the construction of social norms and individual beliefs to the establishment of social institutions. In support of this proposal we argue and present evidence that great apes (and some children with autism) understand the basics of intentional action, but they still do not participate in activities involving joint intentions and attention (shared intentionality). Human children's skills of shared intentionality develop gradually during the first 14 months of life as two ontogenetic pathways intertwine: (1) the general ape line of understanding others as animate, goal-directed, and intentional agents; and (2) a species-unique motivation to share emotions, experience, and activities with other persons. The developmental outcome is children's ability to construct dialogic cognitive representations, which enable them to participate in earnest in the collectivity that is human cognition. PMID- 16262964 TI - Cost-effectiveness of interventions to support self-care: a systematic review. AB - OBJECTIVES: Interventions to support patient self-care of their condition aim to improve patient health and reduce health service costs. Consequently, they have attracted considerable policy interest. There is some evidence of clinical effectiveness but less attention has been paid to whether these interventions are cost-effective. This study examines the quality and quantity of existing evidence of the cost-effectiveness. METHODS: A systematic review was carried out to assess the extent and quality of economic evaluations of self-care support interventions. Thirty-nine economic evaluations were assessed against a quality checklist developed to reflect the special features of these interventions. RESULTS: The majority of the studies claimed that self-care support interventions were cost-effective or cost saving. The overall quality of economic evaluations was poor because of flaws in study designs, especially a narrow definition of relevant costs and short follow-up periods. CONCLUSIONS: The current evidence base does not support any general conclusion that self-care support interventions are cost-effective, but ongoing trials may provide clearer evidence. PMID- 16262965 TI - Reassessment of the cost-effectiveness of hormone replacement therapy in Sweden: results based on the Women's Health Initiative randomized controlled trial. AB - OBJECTIVES: The purpose of the study is to reassess the cost-effectiveness of hormone replacement therapy (HRT) based on new medical evidence found in the Women's Health Initiative (WHI). Within a model framework using an individual state transition model, the cost-effectiveness of 50- to 60-year-old women with menopausal symptoms is assessed based on a societal perspective in Sweden. METHODS: The model has a 50-year time horizon divided into a cycle length of 1 year. The model consists of the following disease states: coronary heart disease, stroke, venous thromboembolic events, breast cancer, colorectal cancer, hip fracture, vertebral fracture, and wrist fracture. An intervention is modeled by its impact on the disease risks during and after the cessation of therapy. The model calculates costs and quality-adjusted life years (QALYs) with and without intervention. The resulting cost per QALY gained is compared with the value of a QALY gained, which is set to SEK 600,000. The model requires data on clinical effects, risks, mortality rates, quality of life weights, and costs valid for Sweden. RESULTS: The cost-effectiveness ratios are estimated at approximately SEK 10,000, which is below the threshold value of cost-effectiveness. On the condition that HRT increases the quality of life weight more than 0.013 units, the therapy is cost-effective. CONCLUSIONS: In conclusion, given the new evidence in WHI, there is still a high probability that HRT is a cost-effective strategy for women with menopausal symptoms. PMID- 16262966 TI - Marginal cost of operating a positron emission tomography center in a regulatory environment. AB - OBJECTIVES: Cost studies of positron emission tomography (PET) imaging are important for resource and operational planning; the most relevant cost analysis in this regard is the marginal cost. Operating within a regulatory environment can add considerably to the costs of providing PET services. Previously published research has not examined the marginal cost structure of PET nor have they described the implications of regulatory compliance to operational costs. The purpose of this study was to conduct a comprehensive cost estimation of PET imaging with 18F-fluorodeoxyglucose (18F-FDG) to better identify the fixed and variable cost components, the marginal cost structure, and the added costs of satisfying regulatory requirements. METHODS: Financial data on capital and operating expenses were collected for the PET center at the Cross Cancer Institute in Edmonton, Alberta, Canada. RESULTS: The total per-service cost for clinical operations ranged between $7,869 (400 annual scans) and $1,231 (3,200 annual scans). The marginal cost for the center remained steady as volume increased up to the throughput capacity. CONCLUSIONS: Results indicate that economies from increased volumes did not arise. Regulatory requirements added significant costs to operating an 18F-FDG-PET center. PMID- 16262967 TI - Physiotherapy after arthroscopic partial meniscectomy surgery: an assessment of costs to the National Health Service, patients, and society. AB - OBJECTIVES: The purpose of this study was to determine and inform clinicians, managers, and budget allocators of the costs incurred to the British National Health Service (NHS), patient, and society when attending clinic-based physiotherapy compared with not attending clinic-based physiotherapy after arthroscopic partial meniscectomy surgery. METHODS: The valuation principle used in this study was the economic concept of opportunity cost. Costs were referred to as direct medical (NHS), direct nonmedical (patient), and indirect (societal) costs. Due to the difficulties of their measurement and valuation, intangible costs, in the form of pain and anxiety related to the effect of receiving or not receiving treatment, have not been considered in this analysis. RESULTS: Providing clinic-based physiotherapy after knee arthroscopic partial meniscectomy surgery is more costly to the NHS and patient, but no more costly to society than when not providing it and does not result in reduced contact with the NHS. CONCLUSIONS: Clinic-based physiotherapy after knee arthroscopic partial meniscectomy surgery is costly and evidence is needed that its effectiveness is high enough to support its use. PMID- 16262968 TI - Quality and relevance of evidence in support of guideline recommendations for sleep laboratory investigations. AB - OBJECTIVES: As part of a review of guidelines for sleep laboratories, this study summarizes the quality and relevance of evidence cited in support of recommendations on investigations for individuals with sleep disorders. METHODS: Quality of evidence in support of each recommendation was rated as A (from well conducted, prospective controlled studies), B (from controlled studies and case series, with minor shortcomings), or C (from case series or case series plus controlled studies with substantial limitations). Relevance of the cited evidence was also rated from A to C, in decreasing order of merit. RESULTS: Of eighty-one recommendations from thirty-seven guidelines, forty-six were supported by evidence from primary studies. Six recommendations were supported by category A evidence, fifteen by category B, and twenty-five recommendations by category C. The cited evidence was highly relevant to eighteen recommendations, of some relevance for twenty-two and of little or no relevance to six. Four recommendations were informed by an absence of available evidence. For the other thirty-one, no evidence was provided in support. CONCLUSIONS: The approach used provided a concise overview of recommendations and supporting evidence for decision-makers. Guidelines on the use of sleep laboratory investigations contain much detailed information, but evidence supporting several recommendations is of limited quality and relevance. Furthermore, good-quality studies of many sleep laboratory applications are needed to assist both health policy formulation and clinical practice. PMID- 16262969 TI - Assessing quality of end-of-life hospital care in a southern European regional health service. AB - OBJECTIVES: During the final period of life, patients with cancer in the Basque Country are given treatment in different types of hospital care. This study compared the quality of care according to the type of care in one of the autonomous communities in Spain. METHODS: A retrospective study was carried out of cancer patients who died in conventional hospital services, home hospitalization services, and palliative care units. In addition to hospital stay and readmission number, variables based on the recommendations of Spanish Society for Palliative Care were studied. RESULTS: End-of-life was diagnosed in 57 percent of a sample of 486 patients, 3 days before death (median). The use of symptom control scales was only documented in the clinical records of eight patients. Sociofamily evaluation was not found. Patients in conventional hospital services were less frequently diagnosed with end-of-life and agony and were significantly different from the rest in the reasons for admission, symptoms assessed, drugs used, administration routes, and dosage forms. Pain was evaluated in 50 percent of the patients and was better controlled in palliative care units. Patients not diagnosed with agony (52 percent) were more frequently not given specific treatment. CONCLUSIONS: End-of-life in cancer patients was diagnosed too late. The quality of care in palliative care units and by home hospitalization service was better than that in conventional hospitalization. Nevertheless, there were areas for improvement in the three modalities of care. PMID- 16262970 TI - Toward a multidimensional assessment of picture archiving and communication system success. AB - OBJECTIVES: Based on a prevalent framework in the information systems field, this study proposes and describes an integrated model for evaluating picture archiving and communication system (PACS) success from multiple users' perspectives. METHODS: Our study details the validation process of the proposed model at a large tertiary-care teaching hospital in Canada. Both qualitative and quantitative data were collected to assess the psychometric properties of the measurement instrument and test the research hypotheses. RESULTS: Our findings clearly reveal that radiologists, technologists, and clinicians have different views regarding the factors influencing PACS success. For instance, the results for radiologists show that their concern with efficiency and productivity is best guaranteed by a system that is reliable and easy to use. Furthermore, that only perceived system usefulness influenced clinicians' satisfaction with PACS is a reflection of the primary impact that technology has on their work, namely, the ability to have instant access to images from any point in the hospital. Even though, overall, all three groups view the adoption of PACS positively, the mean scores indicate that radiologists and technologists seem to be more satisfied and their expectations to be met at a higher level than clinicians. CONCLUSIONS: We believe the measurement instruments developed in this study can be used as a diagnostic tool by project managers interested in better understanding the extent to which different groups of stakeholders perceive the deployment of PACS as being successful and how factors influencing perceptions of PACS success vary across user types. PMID- 16262972 TI - Predicting the impact of new health technologies on average length of stay: development of a prediction framework. AB - OBJECTIVES: The aim of this study was to develop a framework to predict the impact of new health technologies on average length of hospital stay. METHODS: A literature search of EMBASE, MEDLINE, Web of Science, and the Health Management Information Consortium databases was conducted to identify papers that discuss the impact of new technology on length of stay or report the impact with a proposed mechanism of impact of specific technologies on length of stay. The mechanisms of impact were categorized into those relating to patients, the technology, or the organization of health care and clinical practice. RESULTS: New health technologies have a variable impact on length of stay. Technologies that lead to an increase in the proportion of sicker patients or increase the average age of patients remaining in the hospital lead to an increase in individual and average length of stay. Technologies that do not affect or improve the inpatient case mix, or reduce adverse effects and complications, or speed up the diagnostic or treatment process should lead to a reduction in individual length of stay and, if applied to all patients with the condition, will reduce average length of stay. CONCLUSIONS: The prediction framework we have developed will ensure that the characteristics of a new technology that may influence length of stay can be consistently taken into consideration by assessment agencies. It is recognized that the influence of technology on length of stay will change as a technology diffuses and that length of stay is highly sensitive to changes in admission policies and organization of care. PMID- 16262971 TI - Ximelagatran: a new type of oral anticoagulant. AB - OBJECTIVES: This assessment sought to evaluate the comparative benefit and adverse effect profile of ximelagatran, as well as the clinical issues surrounding its potential use. METHODS: We performed a Dialog OneSearch across BIOSIS Previews, EMBASE, MEDLINE, PASCAL, and ToxFile to identify published literature. PubMed and The Cochrane Library were also searched. Gray literature was identified by searching a variety of Web sites of health technology assessment and related agencies and their associated databases. The manufacturer's Canadian office, AstraZeneca, was invited to submit information. RESULTS: Ximelagatran is the first oral agent from a new class of anticoagulants called direct thrombin inhibitors. Other oral anticoagulants require routine blood monitoring; ximelagatran does not. Ximelagatran has been evaluated in the areas of venous thromboembolism management, particularly after orthopedic surgery, and stroke prevention in patients with atrial fibrillation. Overall, ximelagatran's efficacy appears comparable to other anticoagulants in these clinical settings. Also, bleeding rates were generally similar between ximelagatran and comparators but, as for warfarin, bleeding risk increases with higher ximelagatran doses. In addition, there is no specific antidote to help manage ximelagatran-induced bleeding. Finally, significantly more patients exposed to long-term ximelagatran developed elevated liver enzymes more than three times the upper normal limit, compared with patients on comparator anticoagulants. CONCLUSIONS: Given its apparent simplicity of use, ximelagatran carries the potential to replace, at least in part, anticoagulants currently used in the management of venous thromboembolism or for preventing stroke in atrial fibrillation patients. However, the safety of ximelagatran will not be fully known without further evaluation and surveillance for potential liver toxicity. There is also a need to evaluate its use in special populations such as patients with renal failure and patients using several concurrent medications. PMID- 16262973 TI - Time-trends in health technology assessments: an analysis of developments in composition of international health technology assessments from 1989 to 2002. AB - OBJECTIVES: Health Technology Assessment (HTA) as a method for producing evidence in the health-care sector has been used for more than 25 years but has grown in extent during the past years. The objective of this study is to explore a possible evolution in these HTAs, in type of assessed technologies, in type of assessors, and in its methods. METHODS: A structured literature review was conducted of 433 HTA reports from the period 1989 to 2002 by eleven leading HTA institutions worldwide. The review focused on the methodology used, the assessors, and the assessed technologies and was designed to elucidate general time-trends in the practical application of HTA. RESULTS: The study shows that literature reviews are still the most often used method of assessment and accounts for a relatively stable fraction of assessments. The fraction of economic evaluations shows a slightly decreasing trend in contrast to randomized controlled trials and modeling/evaluations, which are applied more frequently. The data also demonstrate a more frequent use of external partners as assessors and a shift between devices and pharmaceuticals as assessed technologies. CONCLUSIONS: The study shows an increase in the number of HTAs but no major developments in assessment methods used and, therefore, no widespread spillover from the development in research methods in general to the field of HTA methodology. PMID- 16262974 TI - Development and application of Model of Resource Utilization, Costs, and Outcomes for Stroke (MORUCOS): an Australian economic model for stroke. AB - OBJECTIVES: To outline the development, structure, data assumptions, and application of an Australian economic model for stroke (Model of Resource Utilization, Costs, and Outcomes for Stroke [MORUCOS]). METHODS: The model has a linked spreadsheet format with four modules to describe the disease burden and treatment pathways, estimate prevalence-based and incidence-based costs, and derive life expectancy and quality of life consequences. The model uses patient level, community-based, stroke cohort data and macro-level simulations. An interventions module allows options for change to be consistently evaluated by modifying aspects of the other modules. To date, model validation has included sensitivity testing, face validity, and peer review. Further validation of technical and predictive accuracy is needed. The generic pathway model was assessed by comparison with a stroke subtypes (ischemic, hemorrhagic, or undetermined) approach and used to determine the relative cost-effectiveness of four interventions. RESULTS: The generic pathway model produced lower costs compared with a subtypes version (total average first-year costs/case AUD$ 15,117 versus AUD$ 17,786, respectively). Optimal evidence-based uptake of anticoagulation therapy for primary and secondary stroke prevention and intravenous thrombolytic therapy within 3 hours of stroke were more cost effective than current practice (base year, 1997). CONCLUSIONS: MORUCOS is transparent and flexible in describing Australian stroke care and can effectively be used to systematically evaluate a range of different interventions. Adjusting results to account for stroke subtypes, as they influence cost estimates, could enhance the generic model. PMID- 16262975 TI - Comparing costs of home- versus hospital-based treatment of infections in adults in a specialist cystic fibrosis center. AB - OBJECTIVES: This study aimed to produce valid patient-based UK National Health Service (NHS) costs for adults with cystic fibrosis to identify differences between hospital- and home-based treatments for infections. METHODS: A costing study was carried out in adults with cystic fibrosis (CF) in the United Kingdom, who required intravenous antibiotic treatments for respiratory infections, administered either at home or in the hospital. The perspective was that of the NHS hospital trust. Data were collected retrospectively for each patient for 1 year using clinical records. Data were collected for 116 adults with CF between 2000 and 2001, when 42,382 treatment days (454 courses) of intravenous antibiotics were administered; 213 courses with intention-to-treat at home and 241 courses with intention-to-treat in the hospital. The mean length of a course was 15.3 days. RESULTS: Patients who had >60 percent of courses at home over 1 year had a mean cost of 13,528 UK pounds, compared with 22,609 pounds for patients who had > 60 percent of courses in the hospital, and a mean cost of 19,927 UK pounds for patients who had an equal mix of home and hospital care (p = .0001). CONCLUSIONS: The key cost-generating events in CF respiratory infections are hospital admissions. Future studies assessing costs should concentrate on factors affecting admissions, length of stay, staff input, and alternative methods of home-care provision, rather than marginal effects, such as using different antibiotics. PMID- 16262976 TI - A test of prospect theory. AB - OBJECTIVES: Prospect theory (PT) hypothesizes that people judge states relative to a reference point, usually assumed to be their current health. States better than the reference point are valued on a concave portion of the utility function; worse states are valued on a convex portion. Using prospectively collected utility scores, the objective is to test empirically implications of PT. METHODS: Osteoarthritis (OA) patients undergoing total hip arthroplasty periodically provided standard gamble scores for three OA hypothetical states describing mild, moderate, and severe OA as well as their subjectively defined current state (SDCS). Our hypothesis was that most patients improved between the pre- and postsurgery assessments. According to PT, scores for hypothetical states previously > SDCS but now < SDCS should be lower at the postsurgery assessment. RESULTS: Fourteen patients met the criteria for testing the hypothesis. Predictions were confirmed for 0 patients; there was no change or mixed results for 6 patients (42.9 percent); and scores moved in the direction opposite to that predicted by PT for 8 patients (57.1 percent). CONCLUSIONS: In general, the direction and magnitude of the changes in hypothetical-state scores do not conform to the predictions of PT. PMID- 16262977 TI - Addressing telehealth's foremost barrier: provider as initial gatekeeper. AB - OBJECTIVES: During the past decade, telehealth has enjoyed a plethora of public funding and publication outlets around the world. Yet, rhetoric appears to be outpacing the actual diffusion and utilization of telehealth technologies for patient care. Several barriers, such as reimbursement and legal/regulatory issues, are commonly cited as impeding the successful deployment of this innovation. However, two separate studies carried out in Michigan that controlled for these barriers point out a more significant initial gatekeeper to the deployment of telehealth, namely providers. METHODS: Multiple data collection strategies were used in both the telehospice and telepsychiatry projects, including utilization logs, surveys, telehospice nursing notes, cost frame data collection, patient interviews, caregiver interviews and focus groups, and videotaped visits. RESULTS: This study summarizes data from the two studies to support the hypothesis that the provider is the most important initial gatekeeper for telemedicine. CONCLUSIONS: The implications from this conclusion have important consequences for health system deployment strategies. Specifically, telemedicine project managers must keep providers' needs (ease of use and incentives) in mind when designing a telemedicine system. PMID- 16262978 TI - Mentoring a developing health technology assessment initiative in Romania: an example for countries with limited experience of assessing health technology. AB - OBJECTIVES: The aim of this study was to assist and facilitate introduction and development of a health technology assessment (HTA) program in Romania. METHODS: Mentoring of an initiative group in Romania was provided by an HTA program in Canada. Mentoring activities included provision of HTA materials, participation in local seminars, facilitating contact with HTA and funding organizations, and in-house training of a professional from Romania. RESULTS: Since 1998, when the relationship was initiated, the Romanian group has been successful in developing an understanding of HTA and awareness of its utility among various decision makers in the health system. Currently, although the need for HTA in Romania exists and interest in developing this activity has been officially expressed, HTA is still early in its development phase. The mentoring support helped to identify and define the need for HTA in Romania. Continuation of the existing relationship can be expected to strengthen the expertise in this country. However, while mentoring has been a valuable activity, it is not, by itself, sufficient to ensure development of an HTA program in Romania. The actions and decisions that could lead to implementing HTA in Romania depend on the local context. CONCLUSIONS: Mentoring services assisted the initiative group in promoting HTA in Romania. The implementation of HTA in Romania has not happened yet, and efforts need to continue to sustain the existing momentum. However, success in implementing an HTA program will depend on essential factors such as local political, economical, and educational support for this initiative and others like it. PMID- 16262979 TI - Six years' experience with interdisciplinary review teams in health technology assessment in Norway. AB - OBJECTIVES: The aim of this study was to evaluate the use of interdisciplinary review teams that have been the main and central work form in making health technology assessments at the Norwegian Centre for Health Technology Assessment. METHODS: Evaluation questionnaires were sent to all 112 participants in the 17 review teams for the period of January 1998 to June 2003 after completion of the literature assessment. Questions were on the theme/mandate of the assessments, composition of the review team, organization of the work, the working method, and update of the report. RESULTS: The teams ranged from 4 to 14 persons regarded as opinion leaders in their field. The project periods lasted from 4 to 33 months. In all, fifty-five participants gave fifty-eight responses (51.8 percent) to the questionnaires. A total of 83 percent thought the theme was well argued, and 62 percent thought the mandate for the assessments was sufficiently clear. Approximately 80 percent were positive to the composition of the review team. In all, 22 percent expressed that the work method was too extensive and 43 percent wanted more tuition. General comments were that the working method gave competence in assessing medical literature, relevant professional training, and tuition in a working method that ensured the legitimacy of their work. CONCLUSIONS: The review team participants were satisfied with most aspects of the work. The Norwegian Centre for Health Technology Assessment will continue using interdisciplinary review teams in making health technology assessments. PMID- 16262980 TI - Cost-effectiveness analysis for priority setting in health: penny-wise but pound foolish. AB - Cost-effectiveness analysis has much conceptual attractiveness in priority setting but is not used to its full potential to assist policy-makers on making choices in health in developed or in developing countries. We call for a shift away from present economic evaluation activities-that tend to produce ad hoc and incomparable economic evaluation studies and, therefore, add little to the compendium of knowledge of cost-effectiveness of health interventions in general toward a more systematic approach. Research efforts in economic evaluation should build on the foundations of cost-effectiveness research of the past decades to arrive at an informative methodology useful for national policy-makers. This strategy means that governments should steer sectoral cost-effectiveness analysis to obtain systematic and comprehensive information on the economic attractiveness of a set of new and current interventions, using a standardized methodology and capturing interactions between interventions. Without redirecting the focus of economic evaluation research, choosing in health care bears the risk to remain penny-wise but pound-foolish. PMID- 16262982 TI - Cognitive and affective neuroscience and developmental psychopathology. PMID- 16262983 TI - Autism at the beginning: microstructural and growth abnormalities underlying the cognitive and behavioral phenotype of autism. AB - Autistic symptoms begin in the first years of life, and recent magnetic resonance imaging studies have discovered brain growth abnormalities that precede and overlap with the onset of these symptoms. Recent postmortem studies of the autistic brain provide evidence of cellular abnormalities and processes that may underlie the recently discovered early brain overgrowth and arrest of growth that marks the first years of life in autism. Alternative origins and time tables for these cellular defects and processes are discussed. These cellular and growth abnormalities are most pronounced in frontal, cerebellar, and temporal structures that normally mediate the development of those same higher order social, emotional, speech, language, speech, attention, and cognitive functions that characterize autism. Cellular and growth pathologies are milder and perhaps nonexistent in other structures (e.g., occipital cortex), which are known to mediate functions that are often either mildly affected or entirely unaffected in autistic patients. It is argued that in autism, higher order functions largely fail to develop normally in the first place because frontal, cerebellar, and temporal cellular and growth pathologies occur prior to and during the critical period when these higher order neural systems first begin to form their circuitry. It is hypothesized that microstructural maldevelopment results in local and short distance overconnectivity in frontal cortex that is largely ineffective and in a failure of long-distance cortical-cortical coupling, and thus a reduction in frontal-posterior reciprocal connectivity. This altered circuitry impairs the essential role of frontal cortex in integrating information from diverse functional systems (emotional, sensory, autonomic, memory, etc.) and providing context-based and goal-directed feedback to lower level systems. PMID- 16262985 TI - An event-related potential study of the impact of institutional rearing on face recognition. AB - Event-related potentials (ERPs) were recorded to brief images of caregivers' and strangers' faces for 72 institutionalized children (IG), ages 7-32 months, and compared with ERPs from 33 children, ages 8-32 months, who had never been institutionalized. All children resided in Bucharest, Romania. Prominent differences in four ERP components were observed: early negative (N170), early positive (P250), midlatency negative (Nc), and positive slow wave (PSW). For all but the P250, the amplitude of these components was larger in the never institutionalized group than the institutionalized group; this pattern was reversed for the P250. Typical effects of the Nc (amplitude greater to stranger vs. caregiver) were observed in both groups; in contrast, the IG group showed an atypical pattern in the PSW. These findings are discussed in the context of the role of experience in influencing the neural circuitry putatively involved in recognizing familiar and novel faces. PMID- 16262986 TI - An event-related potential study of the processing of affective facial expressions in young children who experienced maltreatment during the first year of life. AB - This investigation examined the effects of maltreatment during the first year of life on the neural correlates of processing facial expressions of emotion at 30 months of age. Event-related potentials (ERPs) in response to children passively viewing standardized pictures of female models posing angry, happy, and neutral facial expressions were examined. Four ERP waveform components were derived: early negative (N150), early positive (P260), negative central (Nc), and positive slow wave (PSW). Differences in these waveforms between a group of 35 maltreated and 24 nonmaltreated children were reported. The groups did not differ on the early perceptual negative component (N150), whereas the maltreated children had greater P260 amplitude at frontal leads compared to the nonmaltreated children in response to viewing angry facial expressions. For the Nc component, the nonmaltreated comparison children exhibited greater amplitude while viewing pictures of happy faces compared to angry and neutral faces, whereas the maltreated children showed greater Nc amplitude at central sites while viewing angry faces. For the PSW, the nonmaltreated group showed a greater area score in the right hemisphere in response to viewing angry facial expressions compared to the maltreated group. The results are discussed in terms of brain development and function, as well as their implications for the design and evaluation of preventive interventions. PMID- 16262984 TI - The emergence of the social brain network: evidence from typical and atypical development. AB - Several research groups have identified a network of regions of the adult cortex that are activated during social perception and cognition tasks. In this paper we focus on the development of components of this social brain network during early childhood and test aspects of a particular viewpoint on human functional brain development: "interactive specialization." Specifically, we apply new data analysis techniques to a previously published data set of event-related potential (ERP) studies involving 3-, 4-, and 12-month-old infants viewing faces of different orientation and direction of eye gaze. Using source separation and localization methods, several likely generators of scalp recorded ERP are identified, and we describe how they are modulated by stimulus characteristics. We then review the results of a series of experiments concerned with perceiving and acting on eye gaze, before reporting on a new experiment involving young children with autism. Finally, we discuss predictions based on the atypical emergence of the social brain network. PMID- 16262987 TI - Neurocognitive and electrophysiological evidence of altered face processing in parents of children with autism: implications for a model of abnormal development of social brain circuitry in autism. AB - Neuroimaging and behavioral studies have shown that children and adults with autism have impaired face recognition. Individuals with autism also exhibit atypical event-related brain potentials to faces, characterized by a failure to show a negative component (N170) latency advantage to face compared to nonface stimuli and a bilateral, rather than right lateralized, pattern of N170 distribution. In this report, performance by 143 parents of children with autism on standardized verbal, visual-spatial, and face recognition tasks was examined. It was found that parents of children with autism exhibited a significant decrement in face recognition ability relative to their verbal and visual spatial abilities. Event-related brain potentials to face and nonface stimuli were examined in 21 parents of children with autism and 21 control adults. Parents of children with autism showed an atypical event-related potential response to faces, which mirrored the pattern shown by children and adults with autism. These results raise the possibility that face processing might be a functional trait marker of genetic susceptibility to autism. Discussion focuses on hypotheses regarding the neurodevelopmental and genetic basis of altered face processing in autism. A general model of the normal emergence of social brain circuitry in the first year of life is proposed, followed by a discussion of how the trajectory of normal development of social brain circuitry, including cortical specialization for face processing, is altered in individuals with autism. The hypothesis that genetic-mediated dysfunction of the dopamine reward system, especially its functioning in social contexts, might account for altered face processing in individuals with autism and their relatives is discussed. PMID- 16262988 TI - Love hurts: the evolution of empathic concern through the encephalization of nociceptive capacity. AB - Empathic concern for others is an essential motive for challenges of self regulation at all developmental stages. A child who never develops the capacity for empathic concern may become an ineffective parent, such that developmental psychopathology propagates across generations. We draw on evidence and theory by Panksepp and associates that indicates that infant-mother bonding is mediated by opiate mechanisms. We review the neural systems of pain perception and find these are closely aligned with those for attentional and cognitive self-regulation. Analysis of the limbic and neocortical representations for interpersonal reasoning suggests there are important contributions from visceral, affective, and somatic and cognitive levels. We draw on modern learning theory to propose a critical role for frustration training in development to allow the child to achieve the capacity for tolerating psychological pain that allows effective empathic concern in later relationships. PMID- 16262990 TI - Early adversity and mechanisms of plasticity: integrating affective neuroscience with developmental approaches to psychopathology. AB - Interest in the effects of early adversity on children's development reflects contemporary emphases on early experience in the behavioral sciences and plasticity in the neurosciences. Over the past decade, powerful new tools and approaches for understanding the neural circuitry involved in emotion have become increasingly available. Yet, research in developmental psychopathology has not reaped the full benefits of affective neuroscience approaches and methods. Integration of affective neuroscience approaches can excavate developmental mechanisms, thereby advancing knowledge about the etiology, prevention, and treatment of mental health problems in children. Here, we consider two general principles that can guide understanding of plasticity in the neural circuitry of emotion systems and the development of psychopathology. PMID- 16262989 TI - The circumplex model of affect: an integrative approach to affective neuroscience, cognitive development, and psychopathology. AB - The circumplex model of affect proposes that all affective states arise from cognitive interpretations of core neural sensations that are the product of two independent neurophysiological systems. This model stands in contrast to theories of basic emotions, which posit that a discrete and independent neural system subserves every emotion. We propose that basic emotion theories no longer explain adequately the vast number of empirical observations from studies in affective neuroscience, and we suggest that a conceptual shift is needed in the empirical approaches taken to the study of emotion and affective psychopathologies. The circumplex model of affect is more consistent with many recent findings from behavioral, cognitive neuroscience, neuroimaging, and developmental studies of affect. Moreover, the model offers new theoretical and empirical approaches to studying the development of affective disorders as well as the genetic and cognitive underpinnings of affective processing within the central nervous system. PMID- 16262992 TI - An integrative theory of attention-deficit/ hyperactivity disorder based on the cognitive and affective neurosciences. AB - Attention-deficit/hyperactivity disorder (ADHD) is a behavioral syndrome that arises in early childhood, often co-occurs with conduct disorder and leads, developmentally, to antisocial behavior and substance abuse. Models from cognitive and affective neuroscience have been invoked in an effort to understand the development of ADHD, leading to a broad array of interrelated theories and hypothesized mechanisms. In this paper, we highlight core mechanisms that may cut across several theories and constructs, and thus provide some leverage for further study and conceptualization from a neuroscience perspective. We emphasize the joint operations of frontostriatal and frontocerebellar neural loops in detecting and predicting what and when important events in the environment will occur and their interaction with frontoamygdala loops in assigning emotional significance to these events. We note that weaknesses in the development of these basic operations could lead to decrement in the development of cognitive and affective control and other mental operations mediated by prefrontal cortex during development. In turn, such decrement could lead to many of the phenotypic cognitive and neuropsychological features seen in children with ADHD. PMID- 16262991 TI - A multilevel analysis of cognitive dysfunction and psychopathology associated with chromosome 22q11.2 deletion syndrome in children. AB - We present a multilevel approach to developing potential explanations of cognitive impairments and psychopathologies common to individuals with chromosome 22q11.2 deletion syndrome. Results presented support our hypothesis of posterior parietal dysfunction as a central determinant of characteristic visuospatial and numerical cognitive impairments. Converging data suggest that brain development anomalies, primarily tissue reductions in the posterior brain and changes to the corpus callosum, may affect parietal connectivity. Further findings indicate that dysfunction in "frontal" attention systems may explain some executive cognition impairments observed in affected children, and that there may be links between these domains of cognitive function and some of the serious psychiatric conditions, such as attention-deficit/hyperactivity disorder, autism, and schizophrenia, that have elevated incidence rates in the syndrome. Linking the neural structure and the cognitive processing levels in this way enabled us to develop an elaborate structure/function mapping hypothesis for the impairments that are observed. We show also, that in the case of the catechol-O methyltransferase gene, a fairly direct relationship between gene expression, cognitive function, and psychopathology exists in the affected population. Beyond that, we introduce the idea that variation in other genes may further explain the phenotypic variation in cognitive function and possibly the anomalies in brain development. PMID- 16262995 TI - Reflective thinking and mental imagery: a perspective on the development of posttraumatic stress disorder. AB - Reflective thinking occurs when information stored in long-term memory (LTM) is not sufficient to allow one to respond "automatically" to an object or event. Instead, stored information must be entered into working memory and a novel response or solution produced. In this article I argue that mental imagery plays a central role in this process, and that over the course of normal cognitive development the process of reflective thinking "programs" LTM so that an increasingly large number of tasks can be performed without reflective thinking. Normal cognitive development thus results in a decreasing reliance on imagery. However, if highly emotional images are formed, additional retrieval cues can be entered into LTM, making such images more likely to occur in the future. Such images induce arousal, similar to that induced by the actual event. This line of thinking leads to a novel perspective on the neurocognitive deficits that underlie the development of posttraumatic stress disorder, and may also help to explain some symptoms seen in hyperactivity, impulsiveness, and difficulties in self-control. PMID- 16262996 TI - Applying a cognitive neuroscience perspective to the disorder of psychopathy. AB - Four models of psychopathy (frontal lobe dysfunction, response set modulation, fear dysfunction, and violence inhibition mechanism hypotheses) are reviewed from the perspective of cognitive neuroscience. Each model is considered both with respect to the psychopathy data and, more importantly, for the present purposes, with respect to the broader cognitive neuroscience fields to which the model refers (e.g., models of attention with respect to the response set modulation account and models of emotion with respect to the fear dysfunction and violence inhibition mechanism models). The paper concludes with an articulation of the more recent integrated emotion systems model, an account inspired both by recent findings in affective cognitive neuroscience as well as in the study of psychopathy. Some directions for future work are considered. PMID- 16262993 TI - Attention-deficit disorder (attention-deficit/ hyperactivity disorder without hyperactivity): a neurobiologically and behaviorally distinct disorder from attention-deficit/hyperactivity disorder (with hyperactivity). AB - Most studies of attention-deficit/hyperactivity disorder (ADHD) have focused on the combined type and emphasized a core problem in response inhibition. It is proposed here that the core problem in the truly inattentive type of ADHD (not simply the subthreshold combined type) is in working memory. It is further proposed that laboratory measures, such as complex-span and dual-task dichotic listening tasks, can detect this. Children with the truly inattentive type of ADHD, rather than being distractible, may instead be easily bored, their problem being more in motivation (underarousal) than in inhibitory control. Much converging evidence points to a primary disturbance in the striatum (a frontal striatal loop) in the combined type of ADHD. It is proposed here that the primary disturbance in truly inattentive-type ADHD (ADD) is in the cortex (a frontal parietal loop). Finally, it is posited that these are not two different types of ADHD, but two different disorders with different cognitive and behavioral profiles, different patterns of comorbidities, different responses to medication, and different underlying neurobiologies. PMID- 16262998 TI - Effects of perineural capsaicin treatment of the abdominal vagus on endotoxin fever and on a non-febrile thermoregulatory event. AB - Following perineural capsaicin pretreatment of the main trunks of the abdominal vagus of rats, the first and the second phases of the polyphasic febrile response to intravenous lipopolysaccharide were unaltered, while the third phase of fever course (peak at 5 h) was attenuated. In rats desensitized by intraperitoneal (i.p.) capsaicin (i.e. abdominal non-systemic desensitization), mainly the first but not the later fever phases were reduced. The postprandial hyperthermia to intragastric injection of BaSO4 suspension was attenuated by either i.p. or perineural capsaicin treatment. It is concluded that, in contrast to the accepted model of postprandial hyperthermia, which is mediated by capsaicin-sensitive fibers of the abdominal vagus, in the early phase of polyphasic fever the vagal afferent nerves appear to play no role. The influence of i.p. capsaicin desensitization on this initiating fever phase is independent of the vagus, and a capsaicin-induced alteration of endotoxin action in the liver, prior to vagal nerve endings, is more likely. The late febrile phase is probably influenced by efferent vagal fibers, which might be damaged more easily by perineural than i.p. capsaicin treatment. PMID- 16262994 TI - Neural systems of positive affect: relevance to understanding child and adolescent depression? AB - From an affective neuroscience perspective, the goal of achieving a deeper, more mechanistic understanding of the development of depression will require rigorous models that address the core underlying affective changes. Such an understanding will necessitate developing and testing hypotheses focusing on specific components of the complex neural systems involved in the regulation of emotion and motivation. In this paper, we illustrate these principles by describing one example of this type of approach: examining the role of disruptions in neural systems of positive affect in major depressive disorder in school-age children and adolescents. We begin by defining positive affect, proposing that positive affect can be distinguished from negative affect by its neurobehavioral features. We provide an overview of neural systems related to reward and positive affect, with a discussion of their potential involvement in depression. We describe a developmental psychopathology framework, addressing developmental issues that could play a role in the etiology and maintenance of early-onset depression. We review the literature on altered positive affect in depression, suggesting directions for future research. Finally, we discuss the treatment implications of this framework. PMID- 16262999 TI - The response of ferritin to LPS and acute phase of Pseudomonas infection. AB - Plasma ferritin is an important extracellular iron storage molecule, whose concentration increases drastically in cancer and infection. During infection, the pathogen usurps host iron for its survival and pathogenicity; hence, maintenance of the plasma ferritin level during infection is a crucial host defence mechanism. In this study, the horseshoe crab plasma ferritin complex was purified, characterized, and its involvement in innate immune defence was investigated. The plasma ferritin appears as a 21-kDa subunit on SDS-PAGE. Full length ferritin-H cDNAs (CrFer-H1 and CrFer-H2) were cloned. Analysis of the 5' UTR indicates the existence of a functional iron-response element, suggesting that both the CrFer-H genes may be post-transcriptionally regulated. Northern analysis shows that the CrFer-H is ubiquitously expressed. Within 3 h of lipopolysaccharide challenge, the gene is up-regulated by > 12-fold. In contrast, iron-loading did not result in any significant change. When challenged with Pseudomonas aeruginosa, the plasma ferritin disappeared between 6-48 h and re appeared thereafter, suggesting that during infection, ferritin may be concealed intracellularly as it withholds iron from the invading pathogen. Taken together, these results provide insights into the importance of plasma ferritin as an evolutionarily conserved molecule for the iron-withholding strategy of innate immunity. PMID- 16263000 TI - Polymorphism in IL1B: IL1B-511 association with tuberculosis and decreased lipopolysaccharide-induced IL-1beta in IFN-gamma primed ex-vivo whole blood assay. AB - To determine whether variation in two interleukin 1 family genes (IL1B and interleukin 1 receptor antagonist, IL1RN) is associated with pulmonary tuberculosis (TB), two published polymorphisms at nucleotide positions -511 and +3953 in IL1B and one in the IL1RN 86 bp VNTR were genotyped in 335 smear positive Gambian TB patients, and 298 ethnically matched controls. All individuals were HIV negative. Decreased risk of pulmonary TB was associated with both heterozygosity and homozygosity for the IL1B-511-C allele (OR 0.66, P = 0.027, and OR 0.58, P = 0.015, respectively). Nonetheless, the C allele was present at a frequency of 0.66 in TB cases suggesting that whilst IL-1beta contributes to disease susceptibility, it is not the major factor. There was no association between the IL1B+3953-T/C polymorphism or the 86 bp IL1RN pentallelic repeat and TB in this population. Using an ex-vivo whole blood assay, healthy Gambian individuals who are homozygous for the IL1B-511-T allele failed to exhibit a significant increase in IL-1beta production in response to LPS after IFN-gamma priming. PMID- 16263001 TI - Iso-osmolar prehydration shifts the cytokine response towards a more anti inflammatory balance in human endotoxemia. AB - Clinical experience suggests that the administration of fluids in human endotoxemia reduces symptoms. In the present study, the effects of a standardised fluid protocol on symptoms, inflammatory and hemodynamic parameters in human endotoxemia are determined. With approval of the local ethics committee, 16 healthy volunteers received 2 ng/kg of Escherichia coli endotoxin (O:113). After an overnight fast, nine subjects received 1.5 l of 2.5% glucose/0.45% NaCl the hour prior to the endotoxin administration and 150 ml/h during the course of the experiment ('prehydrated group'). Seven subjects only received a continuous infusion of 75 ml/h during the experiment ('non-prehydrated group'). The course of inflammatory parameters and symptoms were determined and mean arterial pressure, heart rate and forearm blood flow were measured. In the prehydrated group, TNF-alpha increased to 522 +/- 63 pg/ml (mean +/- SEM) while the maximum in the non-prehydrated group was 927 +/- 187 pg/ml (P < 0.04). IL-10 increased similarly in both groups (non-prehydrated 117 +/- 18 pg/ml and prehydrated 99 +/- 18 pg/ml; P = NS). The prehydrated group had a significantly lower (P < 0.004) symptom score and recovered sooner (P = 0.004). Endotoxin-induced changes in hemodynamics revealed no significant differences between groups. We demonstrate that prehydration in experimental human endotoxemia significantly shifts the cytokine balance towards a more anti-inflammatory pattern. This effect is associated with a reduction in symptoms, whereas the changes in hemodynamic parameters are not influenced by prehydration. PMID- 16263002 TI - Conserved mechanisms of signal transduction by Toll and Toll-like receptors. AB - In recent years, considerable progress has been made towards understanding the mechanism by which endotoxin is detected by the cells of the immune system. Lipopolysaccharides are extracted in a soluble form by the serum LPS binding protein and then transferred sequentially to the extrinsic membrane protein CD14 and the co-receptor complex TLR4/MD-2. Our modelling studies suggest that acyl chains of lipid A are buried within the hydrophobic core of MD-2 and this induces cross-linking of the two TLR4/MD-2 complexes, an event that is required to trigger signal transduction. We also propose that, by analogy with the Drosophila Toll receptor, the mechanism of signal transduction is likely to be complex and to involve concerted protein conformational changes. In particular, we propose that receptor-receptor interactions mediated by juxtamembrane sequences play a critical role. PMID- 16263003 TI - Endotoxin: physical requirements for cell activation. AB - Lipopolysaccharide (LPS) is the eminent lipid component of the outer leaflet of the outer membrane of Gram-negative bacteria and the major initiator of innate immune response to bacterial infection. Below the critical micellar concentration (CMC), LPS is exclusively present as a monomer. Above this concentration, aggregates are formed. Increasing the concentration beyond the CMC leads to an increase in aggregate concentration, whereas the concentration of monomers remains constant or even decreases. The question how LPS activates immune cells and whether the aggregate or the monomer is the biologically active unit has been and still is controversial. To prepare clearly defined monomeric solutions, we utilized a dialysis set-up consisting of a donor and an acceptor chamber, separated by a dialysis diaphragm with a cut-off of 5 kDa, thus allowing only monomers to pass. Human mononuclear cells (MNCs) were then stimulated with equal concentrations of aggregates and monomers, respectively, of deep rough mutant LPS from Escherichia coli strain F515 (Re LPS) and TNF-alpha release was determined. In contrast to earlier and very recent work of others, we started with a preparation of aggregate-suspensions and pure monomer-solutions and show that monomers are significantly less active than aggregates in the absence and presence of serum proteins at identical concentrations. In our model, we propose that LPS aggregates are detected by membrane-associated LBP and intercalated into the cell membrane to bring LPS into close proximity to signaling proteins in the membrane, thus finally leading to cell activation. To support this model, we present data showing that LBP is indeed present in or at the cell membrane of human macrophages. PMID- 16263004 TI - Peptidoglycan recognition in innate immunity. AB - The innate immune system recognizes micro-organisms through a series of pattern recognition receptors that are highly conserved in evolution. Peptidoglycan (PGN) is a unique and essential component of the cell wall of virtually all bacteria, is not present in eukaryotes, and is an excellent target for the innate immune system. Indeed, higher eukaryotes, including mammals, have several PGN recognition molecules, including CD14, Toll-like receptor 2 (TLR2), nucleotide oligomerization domain (Nod)-containing proteins, a family of peptidoglycan recognition proteins (PGRPs), and PGN-lytic enzymes (lysozyme and amidase). These molecules induce host responses to micro-organisms, degrade PGN, or have direct antimicrobial effects. PMID- 16263005 TI - Reprogramming of circulatory cells in sepsis and SIRS. AB - Immune status is altered in patients with sepsis or non-infectious systemic inflammatory response syndrome (SIRS). Reduced ex-vivo TNF production by endotoxin-activated monocytes has been regularly reported. This observation is reminiscent of the phenomenon of endotoxin tolerance, and the term 'leukocyte reprogramming' well defines this phenomenon. This review will outline that the hyporesponsiveness of circulating leukocytes is not a generalized phenomenon in sepsis and SIRS. Indeed, the nature of the insult (i.e. infectious versus non infectious SIRS; under anesthesia [surgery] or not [trauma, burn]), the nature of the activator used to trigger leukocytes (i.e. different Toll-like receptor ligands or whole bacteria), the nature of the cell culture (i.e. isolated monocytes versus peripheral blood mononuclear cells versus whole blood assays), and the nature of the analyzed cytokines (e.g. IL-1beta versus IL-1ra; TNF versus IL-10) have a profound influence on the outcome of the response. PMID- 16263006 TI - Complications of vasectomy. AB - INTRODUCTION: Vasectomy is a common method of sterilisation. However, it is less popular than tubal ligation world-wide. It is also a frequent cause of litigation relating to its complications. This article reviews the early and late risks associated with the procedure. PATIENTS AND METHODS: Data collection was done using the internet to search Medline for obtaining evidence-based medicine reviews. Cross-references were obtained from key articles. Websites of government bodies and medical associations were searched for guidelines relating to vasectomy. DISCUSSION: Early complications include haematoma, wound and genito urinary infections, and traumatic fistulae. Vasectomy failure occurs in 0-2% of patients. Late recanalisation causes failure in 0.2% of vasectomies. Significant chronic orchalgia may occur in up to 15% of men after vasectomy, and may require epididyectomy or vasectomy reversal. Antisperm antibodies develop in a significant proportion of men post-vasectomy, but do not increase the risk of immune-complex or atherosclerotic heart disease. Similarly, vasectomy does not enhance risk of testicular or prostate cancer. Vasectomy has a lower mortality as compared to tubal occlusion, but is still significantly high in non industrialised countries because of infections. CONCLUSIONS: Vasectomy, though safe and relatively simple, requires a high level of expertise to minimise complications. Adequate pre-operative counselling is essential to increase patient acceptability of this method of permanent contraception. PMID- 16263007 TI - Controversial topics in orthopaedics: the best bearing couple for hip arthroplasty. PMID- 16263008 TI - Pitfalls in the construction of cancer guidelines demonstrated by the analyses of colorectal referrals. AB - INTRODUCTION: The aim of this study was to develop a system to compare and validate cancer referral guidelines, identifying the pitfalls in their development and provide a mechanism to evaluate their efficacy. PATIENTS AND METHODS: 3302 patients referred from primary care with colorectal symptoms over a 3-year period were assessed. All participants had a comprehensive history obtained via a questionnaire that incorporated all colorectal symptoms. The questionnaires were completed prior to assessment at the hospital. All patients were then assessed at the Colorectal One Stop Clinic (CROSC), underwent investigation and diagnosis achieved. All data were entered into a databank. Current prioritisation guidelines and tools that are used to assess colorectal referrals were applied to this colorectal databank to test their efficacy for cancer detection and referral prioritisation. Sensitivity and specificity for cancer detection and referral rates were assessed. RESULTS: Cancer was detected in 156 patients (4.7%). All prioritisation models (NHS guidelines, Weighted Numerical Score [WNS], Netherlands, Harvard, Mersey, and Somerset) differentiated cancer from non-cancer patients. The use of a few symptoms as risk predictors (e.g. NHS guidelines) causes a decrease in specificity in contrast to a comprehensive risk tool, for example, the WNS at a score of 50 (NHS 54.1%, WNS 62.9%). This results in a significantly higher referral rate (NHS 47.6%, WNS 39.4%) and identifies fewer cancers (NHS 80.1%, WNS 85.9%). Non-evidence based modifications of the NHS guidelines (Somerset and Mersey) caused a further deterioration in specificity, which was reflected in an increased referral rate. Using the WNS, which is objective and a continuous scale, allows adaptation of the referral threshold, balancing sensitivity and specificity to the resources available within a hospital. For example, the WNS of > or = 40 has a sensitivity of 96.8% for cancer detection. CONCLUSIONS: Accurate prospective data collection into a data bank allows testing of referral guidelines as well as providing an adjunct to guideline construction. PMID- 16263009 TI - Complications and mortality following stoma formation. AB - INTRODUCTION: As stoma formation is thought to be declining, we performed a study to evaluate the rate of stoma formation and the impact on stoma complication rates, together with risk factors for complications. PATIENTS AND METHODS: Stoma incidence, individual complications and mortality rates were retrieved from a stoma nurse database of 345 stomas created over an 8-year period. RESULTS: Stoma formation increased over the study period, although the incidence of complications declined. Stoma complications were more frequently seen in emergency surgery. A significant association between stoma complications and mortality was identified. CONCLUSIONS: Age of patient, urgency of surgery and diagnosis were associated with high levels of morbidity and mortality. Stomas are often formed in frail patients unsuitable for anastomosis formation, which may explain the high mortality in ostomy patients. PMID- 16263010 TI - Transanal endoscopic microsurgery--impact on the practice of a colorectal surgeon in a district general hospital. AB - INTRODUCTION: The objective was to assess the impact on the management of colorectal patients treated in a district general hospital within the first year after the introduction of transanal endoscopic microsurgery (TEM). PATIENTS AND METHODS: Data were collected for consecutive unselected patients who underwent TEM. Comparative data were derived from a matched group of patients who underwent anterior resection, peranal procedures (PAR) or transanal resection (TAR) in this unit. RESULTS: Twenty-two patients underwent TEM (11 men and 11 women; aged, 29 87 years; median, 75 years). Eighteen patients had a pre-operative diagnosis of benign rectal neoplasms; three were found to have invasive carcinoma, which might have been missed during TAR. Four patients had a pre-operative diagnosis of rectal cancer and TEM provided local tumour control in three cases. The operating time ranged between 20-150 min (mean, 65 min; median, 57 min). Hospital stay ranged between 0-10 days (mean, 3.7 days; median, 3 days), with a total of 97 in patient days for the entire group of patients. Twenty-four operations were performed (22 TEM and two salvage anterior resections), with an estimated cost of 1544 pounds sterling for consumables used. Alternative treatments in the absence of TEM were considered to involve 10 anterior resections, 5 closures of ileostomy, 30 TAR procedures and one PAR procedure, with an estimated 306 days of in-patient admission, 46 operations and 6245 pounds sterling spent on consumables. CONCLUSIONS: Availability of TEM allows more efficient treatment for a significant number of patients with rectal tumours. The cost of the equipment is offset by a significant decrease in the length of in-patient admissions. PMID- 16263011 TI - Should HIV patients be considered a high risk group for the development of prostate cancer? AB - INTRODUCTION: Only very few case reports exist regarding the incidence of prostate cancer in younger HIV-infected patients. PATIENTS AND METHODS: Two incidences of HIV-infected men diagnosed with prostate cancer, from a cohort of about 200 men treated at St George's Hospital aged 40 years or more are reported. DISCUSSION: On the basis of the evidence presented in both case reports and from the literature, clinicians should be aware that men with HIV infection should be considered a high-risk group for prostate cancer, and consider early PSA screening. PMID- 16263012 TI - Changes in the provision of vascular surgery in a single health region over 10 years. AB - INTRODUCTION: This study examines changes in vascular service provision and activity in a single UK health region over 10 years. PATIENTS AND METHODS: A questionnaire on involvement in vascular surgery was sent to all general surgeons in the Oxford region in 1992, 1997 and 2001. Data on involvement in vascular surgery were obtained from the Department of Health. RESULTS: Over 10 years the number of general surgeons not performing any emergency vascular (arterial) surgery rose from 33% to 65% (P = 0.002). There was also a fall in the number of general surgeons performing only emergency vascular surgery (P = 0.009). Trends were observed towards more vascular emergencies being transferred to another hospital (P = 0.068) and proportionally fewer general surgeons undertaking recurrent varicose vein surgery (P = 0.09). The number of vascular reconstructions was 20.8 per 100,000 population in 1990-1991, rising to 32.3 per 100,000 in 1997-1998 but falling to 27.2 per 100,000 by 1999-2000, the greatest increase in activity was seen in the regional centre. Endovascular procedures increased from 8.2 per 100,000 in 1990-1991 to 21.27 in 1995-1996 falling to 17.4 by 1999-2000. In the regional centre there was a fall of 57% in such procedures from 1996-1997 to 1999-2000. Over 10 years, the overall major amputation rate remained between 10-12 per 100,000. CONCLUSIONS: The changes reflect the gradual separation of vascular surgery from general surgery occurring nationally. They also suggest a more conservative approach in the management of certain vascular conditions. PMID- 16263013 TI - Duplex imaging immediately prior to carotid endarterectomy. AB - INTRODUCTION: In this centre, angiography is used only in selected cases, whilst duplex ultrasound (DU) is the main imaging method prior to carotid endarterectomy (CEA). DU has no associated morbidity and so can be repeated immediately before surgery to detect changes in the carotid plaque or degree of stenosis. PATIENTS AND METHODS: We retrospectively examined our Vascular Surgery Audit database for the last 500 patients admitted for CEA. In each case, the DU scan was repeated immediately before surgery. RESULTS: From 500 admissions, repeat DU immediately prior to surgery detected 8 (1.6%) situations where CEA would no longer have been an appropriate intervention. In four cases, the degree of stenosis was found to be less than 70% on the repeat scan - in three cases the internal carotid artery (ICA) had occluded or sub-occluded and in one case there was a dissection of the ICA plaque. CONCLUSIONS: DU can be repeated, with no associated morbidity, immediately prior to surgery. Such a practice changes management decisions in 1.6% of admissions for CEA, allowing surgery unjustified by current evidence to be avoided. This policy also serves several other important purposes: it is a method of internal validation, provides a means of improving training of vascular technologists and of achieving quality assurance in DU techniques. PMID- 16263014 TI - Blood ordering protocol based on proposed surgical implant in fractured neck of femur patients. AB - INTRODUCTION: Fractured neck of femur patients frequently require blood transfusion. To improve the efficiency of blood ordering, we present a protocol which orders blood specific for the proposed surgical implant. PATIENTS AND METHODS: A retrospective audit over a 1-year period was performed. Patients were divided into six groups dependent on proposed surgical implant. The mean postoperative drop in haemoglobin concentration, the cross-match to transfusion ratio and transfusion indexes were calculated. RESULTS: Statistically significant differences in blood loss were found dependent on implant used. Using guidelines created by the British Committee for Standards in Haematology on the implementation of a maximum surgical blood ordering schedule, a new protocol for blood ordering based on proposed surgical implant was created. CONCLUSIONS: In fractured neck of femur patients awaiting operation, the type of implant can be used to anticipate blood loss and as a guide to blood ordering. PMID- 16263015 TI - The 'ABC' of examining foot radiographs. AB - INTRODUCTION: We report a simple systematic method of assessing foot radiographs that improves diagnostic accuracy and can reduce the incidence of inappropriate management of serious forefoot and midfoot injuries, particularly the Lisfranc type injury. STUDY GROUP AND METHODS: Five recently appointed senior house officers (SHOs), with no casualty or Orthopaedic experience prior to their appointment, were shown a set of 10 foot radiographs and told the history and examination findings recorded in the casualty notes of each patient within 6 weeks of taking up their posts. They were informed that the radiographs might or might not demonstrate an abnormality. They were asked to make a diagnosis and decide on a management plan. The test was repeated after they were taught the 'ABC' method of evaluating foot radiographs. RESULTS: Diagnostic accuracy improved after SHOs were taught a systematic method of assessing foot radiographs. The proportion of correct diagnoses increased from 0.64 to 0.78 and the probability of recognising Lisfranc injuries increased from 0 to 0.6. CONCLUSIONS: The use of this simple method of assessing foot radiographs can reduce the incidence of inappropriate management of serious foot injuries by casualty SHOs, in particular the Lisfranc type injury. PMID- 16263016 TI - MRSA--why treat the symptoms and not the disease? AB - The recent debate over MRSA in our community is really getting to the state of the ridiculous. There is no question that this bacterium, which has been around for at least 40 years, is becoming a bigger and bigger menace in hospitals. Prior to the election, the shadow health secretary, Andrew Lansley, claimed that hospitals are being told to push more patients through beds rather than concentrating on hygiene and this is the cause of the epidemic. He calls for a search and destroy strategy to clean up wards. The health secretary, Dr John Reid, blames the increased use of contract cleaners under the last Tory government for the rise in the rates of infection. This is supported by the Public Sector Trades Union. They believe that it is the high input of patients that prevents MRSA and other hospital-acquired infections being tackled effectively. The answer, they feel, is strict hospital hygiene and frequent hand washing, with a higher proportion of single rooms.Now, we are told that matrons should take charge of cleanliness in hospitals. Despite all the hand wringing and washing, the morbidity and mortality are both in an upward spiral. PMID- 16263017 TI - Use of the CRABEL Score for improving surgical case-note quality. AB - INTRODUCTION: Quality assurance of medical record keeping in general surgery is facilitated by use of the CRABEL Score. Critical appraisal and constant feedback to staff plays an important part in improving case-note quality. MATERIALS AND METHODS: For each case-note audit, a house officer reviewed two sets of case notes for each of six consultant surgeons. Scores were awarded according to initial clerking, subsequent entries, consent, and discharge summary. Overall scores were derived by subtracting deductions for omissions in each category from a starting score of 100. A larger number of points deducted due to absent data leads to a lower overall score and indicates poorer quality case notes. After four audits, a clerking proforma specifically designed to address some of the common areas of weakness identified in our record keeping was introduced and a further audit was performed in March 2004 to assess its impact. RESULTS: The mean score was lowest in the September 2001 audit and improved over the next two audits. However, there was a small reduction in September 2003 compared to September 2002. When the individual sections of the score were looked at separately, the greatest contribution to a poor score comes from the 'subsequent entries' section since there are five entries scored individually leading to a cumulative effect on the overall score. Within both the 'initial clerking' and 'subsequent entries' sections, early audits showed poor performance across a range of areas but consistent poor implementation of the guidelines was seen in a small number of specific areas as record keeping improved. The quality of medical notes improved over the first three cycles but the improvement was not maintained subsequently. DISCUSSION: The CRABEL score has been shown to be a useful, reproducible and easy-to-perform objective assessment of the quality of medical record keeping. Repeated audit cycles have ensured that case-note quality remains a high priority and have also led to the development of standardised admission documentation. Introduction of the latter has led to a measurable improvement in medical record keeping. PMID- 16263018 TI - Quality of clinical case note entries: how good are we at achieving set standards? AB - INTRODUCTION: High quality entries in case notes are becoming increasingly important. Standards exist on what information entries should contain. We have compared case notes from surgical teams at the Royal Glamorgan Hospital with standards based on guidelines from The Royal College of Surgeons of England. PATIENTS AND METHODS: A total of 120 case notes, randomly selected from the department of general surgery, were reviewed. RESULTS: An 80% compliance was achieved in 25/35 standards and 100% was achieved in 6 (patient's name, date, surgeon's name and type of operation on the operation sheet and consent form signed and dated). The following fell short of 80% compliance: PAS number on every page (75%); entries timed (27%); and clinician's name (16%) and designation (27%) printed. Social history was only recorded in 73% of clerkings and family history in 33%. Results of laboratory tests were signed in 65% of notes and radiological tests were signed in 41%. CONCLUSIONS: Healthcare professionals need to be aware of, and comply with, standards. House officers should be given information about standards at departmental induction or during medical training. PMID- 16263019 TI - Alveolar bone grafting: achieving the organisational standards determined by CSAG, a baseline audit at the Birmingham Children's Hospital. AB - INTRODUCTION: Birmingham Children's Hospital (BCH) is the centre for a regional comprehensive cleft service attempting to implement the national guidelines for minimum standards of care. A national audit of cleft management (CSAG) found that 58% of alveolar bone grafts were successful; published series suggest that success rates can be of the order of 95%. We present the results of an audit of alveolar bone grafting over a 33-month period, after implementation. PATIENTS AND METHODS: A retrospective clinical process audit was taken from the hospital notes and an analysis of radiological outcome by Bergland score was obtained by two independent assessors. RESULTS: The audit highlighted the difficulties of integrating the increased clinical workload. Other difficulties included poorly standardised pre- and postoperative occlusal radiography, inconsistent orthodontic management and a lack of prospective data collection. An 81% success rate for alveolar bone grafting compares favourably to the CSAG study. Of 82 patients, 68 had sufficient data for a retrospective review; 21 were our own patients and 47 were referred into the centralised service. The success of bone grafting as defined by CSAG (including Bergland scores) is based on only two thirds of the patients as many have their orthodontic treatment managed in more distant units and radiographs are much harder to obtain. Bone grafting later than age 11 years, was true for 28% (6/21) of our BCH patients and 46% (22/47) for those referred to our service. CONCLUSIONS: This audit demonstrates what has been achieved in a re-organised service in the context of Real Politik in the NHS and suggests the areas that require improvement. PMID- 16263020 TI - Unplanned admissions following ambulatory plastic surgery--a retrospective study. AB - INTRODUCTION: Admission for overnight or longer hospital stay from a day-case unit is an unwelcome event. This audit was designed to identify the incidence of unplanned admissions and also to detect the potential factors for such overstays. PATIENTS AND METHODS: Details of plastic surgical day-case patients were obtained from the hospital records for a period of 6 months (February 2002 to July 2002). RESULTS: The total number of unplanned admissions was 28 (3.55%). High rates were recorded in patients above the age of 80 years, male patients and body mass index > 30 kg/m2. Duration of surgery (> 45 min) and waiting time in the day surgery unit (> 2.5 h) had significant correlation with the overstays. Grade of the surgeon was not an important determinant factor. Of procedures which resulted in an unexpected admission, the most frequent were otoplasty (8.4%; n = 71) and fasciectomy for Dupuytren's contracture (14.5%; n = 41). Unexpected admissions were also high following rhinoplasty and nipple reconstruction but the total number of operations performed were not significant (n = 12 in each group). Overall, the cause of the overstays were mostly surgical (71.4%) followed by anaesthetic (28.5%) and social (7.1%). DISCUSSION: With an overall unplanned admission rate of 3.55%, our unit is close to the national standard of 2-3%, as advocated by The Royal College of Surgeons of England. Postoperative bleeding (60%) was the most important surgical reason for overstays followed by intravenous antibiotics, wound drainage, excessive duration of the procedure and additional, unplanned procedures. Among the anaesthetic factors, postoperative pain was the leading cause (62.5%) followed by nausea, vomiting and adverse anaesthetic reaction. CONCLUSIONS: The reasons for unplanned admissions are multifactorial and merit appropriate patient selection and proper estimation of the disease process. PMID- 16263021 TI - Routine magnetic resonance cholangiopancreatography and intra-operative cholangiogram in the evaluation of common bile duct stones. AB - INTRODUCTION: Magnetic resonance cholangiopancreatography (MRCP) is a safe and sensitive investigation for the imaging of common bile duct pathology. When used to exclude common bile duct (CBD) stones, MRCP may obviate the need for intra operative cholangiogram (IOC). In this prospective study, we looked at the single centre results of patients who underwent cholecystectomy with IOC following pre operative MRCP. PATIENTS AND METHODS: Over a period of 18 months, 69 patients (24 male and 45 female; mean age 59 years [range, 19-86 years]) were investigated by MRCP prior to cholecystectomy. All patients underwent IOC. Inclusion criteria for MRCP consisted of derangement of liver function tests and/or history of jaundice in cases of ultrasound-proven cholelithiasis. RESULTS: Sixteen patients had suspected stones or filling defects on MRCP; all but two of these were confirmed to be stones on IOC. In only one patient was a stone visualised on IOC and not seen on MRCP. CONCLUSION: MRCP may be the only pre-operative investigation needed for exclusion of CBD stones, obviating the necessity for IOC. PMID- 16263022 TI - Metachronous stomal adenocarcinoma following abdominoperineal resection for rectal cancer. AB - Although metachronous colorectal tumours are relatively common, they seldom occur at stomasites. We present the case of a 57-year-old woman who developed a colostomy site malignancy. Possible associations and risk factors are discussed. PMID- 16263023 TI - Day-case breast cancer axillary surgery--more data needed. PMID- 16263025 TI - Management of gall stone pancreatitis. PMID- 16263026 TI - A Caribbean perspective. PMID- 16263027 TI - Computerised databases. PMID- 16263028 TI - Transanal resection of the prostate. PMID- 16263029 TI - Extrapulmonary tuberculosis. PMID- 16263030 TI - Google versus PubMed. PMID- 16263031 TI - Fracture liaison service. PMID- 16263032 TI - Inguinal orchidectomy. PMID- 16263034 TI - Health education from 1775 to 2005. PMID- 16263036 TI - Transforming public health through community partnerships. PMID- 16263037 TI - When chronic conditions become acute: prevention and control of chronic diseases and adverse health outcomes during natural disasters. PMID- 16263035 TI - Old black water. PMID- 16263038 TI - At the cutting edge or the center of the storm? Innovation in public health through health promotion and education in state health departments. PMID- 16263039 TI - Costs and savings associated with community water fluoridation programs in Colorado. AB - INTRODUCTION: Local, state, and national health policy makers require information on the economic burden of oral disease and the cost-effectiveness of oral health programs to set policies and allocate resources. In this study, we estimate the cost savings associated with community water fluoridation programs (CWFPs) in Colorado and potential cost savings if Colorado communities without fluoridation programs or naturally high fluoride levels were to implement CWFPs. METHODS: We developed an economic model to compare the costs associated with CWFPs with treatment savings achieved through averted tooth decay. Treatment savings included those associated with direct medical costs and indirect nonmedical costs (i.e., patient time spent on dental visit). We estimated program costs and treatment savings for each water system in Colorado in 2003 dollars. We obtained parameter estimates from published studies, national surveys, and other sources. We calculated net costs for Colorado water systems with existing CWFPs and potential net costs for systems without CWFPs. The analysis includes data for 172 public water systems in Colorado that serve populations of 1000 individuals or more. We used second-order Monte Carlo simulations to evaluate the inherent uncertainty of the model assumptions on the results and report the 95% credible range from the simulation model. RESULTS: We estimated that Colorado CWFPs were associated with annual savings of 148.9 million dollars (credible range, 115.1 million dollars to 187.2 million dollars) in 2003, or an average of 60.78 dollars per person (credible range, 46.97 dollars dollars to 76.41 dollars). We estimated that Colorado would save an additional 46.6 million dollars (credible range, 36.0 dollars to 58.6 dollars million) annually if CWFPs were implemented in the 52 water systems without such programs and for which fluoridation is recommended. CONCLUSION: Colorado realizes significant annual savings from CWFPs; additional savings and reductions in morbidity could be achieved if fluoridation programs were implemented in other areas. PMID- 16263040 TI - Perceived likelihood of developing diabetes among high-risk Oregonians. AB - INTRODUCTION: Prevention of diabetes in people at highest risk for developing the disease is an important public health opportunity, considering the disease's increasing prevalence, its devastating impact on health and its high economic cost, the availability of efficacious and cost-effective treatments to reduce complications, and recent evidence that it can be delayed or prevented with lifestyle interventions. METHODS: The Oregon Diabetes Prevention and Control Program collected and analyzed responses from a statewide telephone survey conducted in 2003 to determine whether Oregon adults at highest risk for diabetes 1) believed that they were at risk for developing diabetes in the future, 2) had talked with a health care professional about diabetes, and 3) had been tested for the disease. Pearson chi-square tests and logistic regression analyses were conducted to identify independent associations of select characteristics with the study factors of interest. RESULTS: Even among respondents at highest risk for developing diabetes, at most one third reported being concerned about developing diabetes, one fifth reported having discussed their risk with a health professional in the previous year, and less than half reported having been tested for diabetes by a health provider in the previous year. After adjusting for multiple factors, we found that having a family history of diabetes was consistently associated with perceived risk of developing diabetes, discussion about diabetes with a health professional, and diabetes testing. CONCLUSION: Many Oregon adults at high risk for developing diabetes are unconcerned about their risk for developing the disease, and few have discussed their risk of diabetes with a health professional. Findings from this study suggest the need for increased recognition of future diabetes risk by high-risk individuals and health professionals to help translate diabetes prevention into practice. PMID- 16263042 TI - From concept to practice: using the School Health Index to create healthy school environments in Rhode Island elementary schools. AB - INTRODUCTION: The prevalence of childhood obesity is increasing, and schools are ideal places to support healthy eating and physical activity. In 2000, the Centers for Disease Control and Prevention (CDC) developed the School Health Index, a self-assessment and planning tool that helps schools evaluate and improve physical activity and nutrition programs and policies. Although many state education agencies, health departments, and individual schools have used the School Health Index, few systematic evaluations of the tool have been performed. We examined the physical activity and nutrition environments in Rhode Island's public elementary schools with high and low minority student enrollments and evaluated a school-based environmental and policy intervention that included implementation of the School Health Index. METHODS: As part of a CDC Division of Nutrition and Physical Activity cooperative agreement awarded to the Rhode Island Department of Health, we conducted a needs assessment of 102 elementary schools and implemented an intervention in four inner-city elementary schools. In phase 1, we analyzed the Rhode Island Needs Assessment Tool (RINAT), a telephone survey of principals in approximately 50% of all Rhode Island public elementary schools in the state during the 2001-2002 school year (n = 102). Comparisons of the nutrition and physical activity environments of schools with low and high minority enrollment were calculated by cross-tabulation with the chi-square test. In phase 2, we used process and outcome evaluation data to assess the use of the School Health Index in creating healthier environments in schools. Our intervention--Eat Healthy and Get Active!--involved implementing three of the eight School Health Index modules in four Rhode Island elementary schools. RESULTS: Survey data revealed that schools with high minority enrollment (student enrollment of > or =10% black, > or =25% Hispanic, or both) offered few programs supporting healthy eating and physical activity (P < .05). Schools with high and low minority enrollment both offered nonnutritious foods and beverages. Process evaluation data revealed that 1) principals play a pivotal role on School Health Index teams, 2) schoolwide validation of a team's small successes is crucial for sustaining a commitment to healthy lifestyle policies and programs, and 3) external facilitators are essential for implementation success. Outcome data showed that all schools developed at least one policy or environmental strategy to create a healthy school environment. Only two schools implemented immediate changes. CONCLUSION: Needs assessment, external facilitation, and evaluation are the foundation for sustainable school-based policies. Although the School Health Index is universally perceived as a user-friendly assessment tool, implementation is likely to be less successful in schools with low staff morale, budgetary constraints, and inconsistent administrator support. PMID- 16263041 TI - Using paid radio advertisements to promote physical activity among Arkansas tweens. AB - INTRODUCTION: The level of physical activity among children is a growing concern. Evidence shows that many children aged 9 to 13 years (tweens) do not participate in any organized physical activity during their nonschool hours, and some do not engage in any free-time physical activity. Physical inactivity is associated with a host of chronic diseases such as diabetes and cardiovascular disease. Paid media advertisements have been an effective method of promoting physical activity. METHODS: From March 10, 2003, through June 29, 2003, we aired paid radio advertisements in six major Arkansas metropolitan areas to promote physical activity among tweens. In September 2003, we surveyed 295 Arkansas tweens by telephone to assess their exposure to the advertisements and the impact of the advertisements on their intent to participate in physical activity. In the same telephone survey, we also asked questions about the respondents' physical activity level. The data were weighted so that the results would be representative of the areas surveyed. The statistical analysis was performed using SPSS, version 11.5 (SPSS Inc, Chicago, Ill). RESULTS: Of the tweens surveyed, 56.4% (95% confidence interval [CI], 50.7%-62.1%) reported hearing the radio advertisements. Of the tweens who heard the advertisement messages, 76.1% (95% CI, 69.4%-82.8%) said the messages made them more likely to get involved in physical activity. Younger tweens (aged 9 and 10 years) were less likely to have heard the advertisements than older tweens (aged 11 to 13 years). However, the advertisements were more likely to cause younger tweens to want to get involved in physical activity (odds ratio [OR] = 6.89, P = .003) than older tweens. Of the tweens surveyed, 74.9% (95% CI, 70.0%-79.8%) reported that they were involved in nonschool-sponsored sports, and 45.3% (95% CI, 39.6%-51.0%) were involved in school-sponsored sports. CONCLUSION: Paid media advertisements may be an effective way to promote physical activity among tweens. More rigorous evaluations of the impact of paid media advertisements among tweens, with larger samples and additional media markets, are needed. Future physical activity promotion efforts should focus on encouraging participation in school-sponsored sports and creating community environments conducive to physical activity. PMID- 16263043 TI - Statewide community-based health promotion: a North Carolina model to build local capacity for chronic disease prevention. AB - BACKGROUND: Public health faces major challenges to building state and local infrastructure with the capacity to address the underlying causes of chronic disease. We describe a structured statewide approach to providing technical assistance for local communities to support and develop health promotion capacity. CONTEXT: Over the last two decades, the North Carolina Statewide Health Promotion program has supported local approaches to the prevention and control of chronic disease. In 1999, a major change in the program required local health departments to focus on policy-change and environmental-change strategies for addressing three major risk factors: physical inactivity, poor diet, and tobacco use. METHODS: State program consultants provided technical assistance and training opportunities to local programs on effective policy-change and environmental-change strategies and interventions, based on needs defined by a statewide monitoring and evaluation system. CONSEQUENCES: The percentage of health departments in North Carolina with interventions addressing at least one of three targeted risk factors in 2004 approached 100%; in 2001, this percentage was 62%. Additionally, between 2001 and 2004, the number of health departments reporting policy or environmental outcomes related to these risk factors almost doubled. INTERPRETATION: Requiring local programs to implement policy-change and environmental-change interventions that address the three major behavioral risk factors provides an organized framework for accountability. An established reporting system guides technical assistance efforts and monitors their effectiveness based on standardized objectives that address the full scope of the socioecologic model. PMID- 16263044 TI - Implementing a new diabetes resource for Wisconsin schools and families. AB - BACKGROUND: Diabetes is one of the most common diseases in the nation. Students with diabetes face the daily task of balancing food, physical activity, and medication to survive. Teachers and school personnel often lack the knowledge needed to assist them. CONTEXT: An estimated 2647 schoolchildren in Wisconsin have diabetes. The Wisconsin Diabetes Prevention and Control Program frequently receives anecdotal reports from parents and diabetes educators on the care of children with diabetes in the schools; the program also manages requests for information on new diabetes-related equipment from school personnel. METHODS: A statewide workgroup convened to develop Children with Diabetes: A Resource Guide for Wisconsin Schools and Families, aimed at improving the school staff's knowledge of diabetes and its management and their awareness of the benefits of maintaining glucose control. Training sessions for school professionals were developed and conducted around the state. All attendees were asked to complete an evaluation of the training. In addition, the workgroup included an evaluation form with each guide distributed and conducted a follow-up survey on the impact of the guide and changes to school policies. CONSEQUENCES: Of the 762 people who attended training sessions, 631 (83%) completed the evaluation form. On questions about the training session's content, quality, organization, and appropriateness, responses averaged 4.42 points on a scale of 1 (poor) to 5 (excellent). More than 9713 resource guides were distributed to more than 1359 individuals; 58 recipients responded to the evaluation form included with the resource guide, with 57 (98%) of these indicating that they would recommend the guide to others. Preliminary results of the follow-up impact survey show that many positive changes have been implemented to improve the school environment for children with diabetes since the resource guide was implemented. INTERPRETATION: This model of working with school professionals, health care practitioners, parents, and community organizations to create a resource guide with accompanying training sessions can be used in other states to accomplish similar goals of increasing knowledge about diabetes and improving social and policy environments. PMID- 16263045 TI - Collaboration between Oregon's chronic disease programs and Medicaid to decrease smoking among Medicaid-insured Oregonians with asthma. AB - BACKGROUND: Environmental tobacco smoke is a leading environmental asthma trigger and has been linked to the development of asthma in children and adults. Smoking cessation and reduced exposure to secondhand tobacco smoke are key components of asthma management. We describe a partnership involving two state agencies and 14 health plans; the goal of the partnership was to decrease smoking and exposure to environmental tobacco smoke among Medicaid-insured Oregonians with asthma. CONTEXT: Oregon's asthma rate is higher than that of the national population, and approximately one third of Oregonians with asthma smoke. The Health Promotion and Chronic Disease Prevention Program (HPCDP) in the Oregon Department of Human Services has collaborated with the Office of Medical Assistance Programs (OMAP) to promote preventive care at the population level. METHODS: Two HPCDP programs- the Oregon Asthma Program and the Oregon Tobacco Prevention and Education Program -worked with OMAP to launch the statewide Asthma-Tobacco Integration Project in 2003. A primary focus of the project is the development of partnerships among health plans, health care providers, and large health care organizations to integrate asthma management and smoking control through systems innovations and provider education. OMAP and its participating health plans also decided to focus cessation efforts on its members with chronic diseases. In addition, HPCDP has collaborated with OMAP to distribute educational tools and information about tobacco's impact on asthma morbidity to Oregon's health care providers who serve low-income Oregonians. CONSEQUENCES: The partnership between OMAP and HPCDP program staff members has allowed them to discuss problems, leverage resources, and obtain support for many public health initiatives. In addition, OMAP-HPCDP collaboration on educational workshops and outreach to health care providers has helped convince quality improvement specialists and administrators about the importance of addressing smoking among patients with asthma. The Asthma-Tobacco Integration Project has also led to formative research aimed at increasing community involvement in promoting tobacco-free environments. INTERPRETATION: Collaboration between HPCDP and OMAP has been an important factor in Oregon's successful smoking cessation efforts in general and in recent efforts to address tobacco use among Oregonians with asthma. PMID- 16263046 TI - Texas' community health workforce: from state health promotion policy to community-level practice. AB - BACKGROUND: Imagine yourself in Texas as a newly arrived immigrant who does not speak English. What would you do if your child became ill? How would you find a doctor? When you find one, will the doctor speak your native language or understand your culture? In a state of approximately 22 million people, many Texas residents, marginalized by poverty and cultural traditions, find themselves in this situation. To help them, some communities across Texas offer the services of promotores, or community health workers, who provide health education and assist with navigating the health care system. CONTEXT: In 1999, Texas became the first state in the nation to recognize these workers and their contributions to keeping Texans healthy. This paper examines a state health promotion policy that culminated in a training and certification program for promotores and the impact of this program on the lay health education workforce in Texas. METHODS: In 1999, the Texas legislature established the 15-member Promotor(a) Program Development Committee to study issues involved in developing a statewide training and certification program. During its 2-year term, the committee met all six of its objectives toward establishing and maintaining a promotor(a) certification program. CONSEQUENCES: By the end of December 2005, it is estimated that there will be more than 700 certified promotores in Texas. State certification brings community health workers into the public health mainstream as never before. INTERPRETATION: Promotores, a community health safety net and a natural extension of the health and human services agencies, improve health at the neighborhood level. Certification brings renewed commitment to serving others and a distinction to those who have been the unsung heroes of public health for decades. PMID- 16263047 TI - The vending and a la carte policy intervention in Maine public high schools. AB - BACKGROUND: A healthy school nutrition environment may be important for decreasing childhood overweight. This article describes a project to make healthier snacks and beverages available in vending machines and a la carte programs in Maine public high schools. CONTEXT: Seven public high schools in Maine volunteered to participate in this project. Four schools made changes to the nutrition environment, and three schools that served as controls did not. The nutrition guidelines were to offer only low-fat (not more than 30% of total calories from fat) and low-sugar (not more than 35% by weight of sugar) items in vending machines and a la carte programs. METHODS: Strategies to implement the project included early communications with school officials, monetary stipends for participation, identification of a school liaison, and a committee at each school to promote the healthy changes. Baseline nutrient content and sales of all competitive foods and beverages were assessed to develop the guidelines for changes in the four schools. Student volunteers at all seven schools were measured for height, weight, diet quality, and physical activity level to assess the impact of the change to the nutrition environment. Baseline measures were taken in the spring semester of 2004. Nutrition changes were made to the a la carte programs and vending machines in the four intervention schools at the start of the fall semester of 2004. Follow-up nutrition assessment and student data collection occurred in the spring semester of 2005. CONSEQUENCES: Healthy changes in vending machines were more easily achieved than those made in the a la carte programs. Technical assistance and ongoing support were essential for successful implementation of this intervention. INTERPRETATION: It is possible to improve the nutrition environment of Maine public high schools. Stakeholder support is essential to sustain healthy changes. PMID- 16263048 TI - Engaging rural youth in physical activity promotion research in an after-school setting. AB - BACKGROUND: West Virginia, the second most rural state in the nation, has a higher than average prevalence of chronic diseases, especially those related to physical inactivity and obesity. Innovative educational approaches are needed to increase physical activity among adults and youth in rural areas and reduce rural health disparities. This paper describes West Virginia's Health Sciences and Technology Academy (HSTA) Education and Outreach on Healthy Weight and Physical Activity. The project involved teachers and underserved high school students in social science research aimed at increasing physical activity among student and community participants. CONTEXT: The HSTA is an ongoing initiative of university school-community partnerships in West Virginia that offers academic enrichment to high-school students in after-school clubs. For this project, six HSTA clubs were awarded grants to conduct research on physical activity promotion during the 2003 2004 school year. The project was funded by the Centers for Disease Control and Prevention. METHODS: Focus groups, workshops, and targeted technical assistance were used to assist teachers and students with developing, implementing, and evaluating their research projects. Each club completed one project, and students reported on their research at the annual HSTA symposium held in the spring. Teachers documented their experience with the projects in process journals before and during implementation. CONSEQUENCES: Data from the teachers' process journals revealed that they believed this research experience increased their students' interest in health and health science careers and increased their students' understanding of social science research methods. Challenges included lack of time after school to complete all activities, competing student activities, limited social science research experience of both teachers and students, and delays that resulted from a lengthy human subjects approval process. INTERPRETATION: The entire process was too ambitious to be achieved in one school year. Recommendations for future implementation include offering training modules on social science research methods for both teachers and students. These modules could be offered as a graduate course for teachers and as an in-school elective within the curriculum or as a summer institute for students. This preparatory training might alleviate some of the time management issues experienced by all the projects and could result in more skilled teacher and student researchers. PMID- 16263050 TI - Geocoding and social marketing in Alabama's cancer prevention programs. AB - The Alabama Department of Public Health (ADPH) is collaborating with the National Cancer Institute to develop detailed profiles of underserved Alabama communities most at risk for cancer. These profiles will be combined with geocoded data to create a pilot project, Cancer Prevention for Alabama's Underserved Populations: A Focused Approach. The project's objectives are to provide the ADPH's cancer prevention programs with a more accurate and cost-effective means of planning, implementing, and evaluating its prevention activities in an outcomes-oriented and population-appropriate manner. The project links geocoded data from the Alabama Statewide Cancer Registry with profiles generated by the National Cancer Institute's cancer profiling system, Consumer Health Profiles. These profiles have been successfully applied to market-focused cancer prevention messages across the United States. The ADPH and the National Cancer Institute will evaluate the efficacy of using geocoded data and lifestyle segmentation information in strategy development and program implementation. Alabama is the first state in the nation not only to link geocoded cancer registry data with lifestyle segmentation data but also to use the National Cancer Institute's profiles and methodology in combination with actual state data. PMID- 16263049 TI - Energizing community health improvement: the promise of microgrants. AB - BACKGROUND: The Healthy Carolinians community microgrants project provided microgrants to community-based organizations (CBOs) across North Carolina. These grants were made to serve as a catalyst to engage the CBOs in health promotion activities that addressed Healthy People 2010 objectives. The purpose of this initiative was to increase the awareness of Healthy People 2010 objectives, mobilize resources, and create new partners in community health improvement. CONTEXT: In 1993, Healthy Carolinians, a statewide network of public-private partnerships, was established at the county level to address North Carolina's health objectives that aligned with national Healthy People 2010 objectives. This network of Healthy Carolinians partnerships provided the vehicle for distributing the microgrants. METHODS: Funding was distributed to 32 Healthy Carolinians partnerships that, in turn, awarded 199 microgrants (2010 dollars each) to CBOs to address state and national health objectives. Each CBO selected its own objectives based on Healthy People 2010 objectives and designed its own interventions. Surveys of the CBO project managers and final reports were used for evaluation. A survey of the Healthy Carolinians partnership coordinators provided additional insight. CONSEQUENCES: Of the 199 surveys mailed to CBOs, 153 (77%) responded. Nearly half (43.7%) of the microgrants were used to focus on three major health risk factors: 27.1% on physical activity and fitness, 13.1% on nutrition and overweight, and 3.5% on tobacco use. At the end of the project, 96.1% of the respondents reported that they were familiar with the Healthy People 2010 objectives. Final reports showed that an estimated 52,739 hours of CBO staff and volunteer time were contributed to microgrant programs. All Healthy Carolinians partnership coordinators responded to a survey; 100% stated that they had new access to priority populations within their community. INTERPRETATION: The Healthy Carolinians microgrant project demonstrated a cost-effective, alternative approach to funding community-based health promotion and injury control activities. This model was decentralized, so it empowered communities and CBOs to be responsible for community health improvement. Public health professionals with limited funds should consider this alternative approach, which mobilized existing community organizations and effectively addressed national and state health objectives. PMID- 16263051 TI - Progress in the genetics of progressive supranuclear palsy: tau gene and beyond. PMID- 16263052 TI - A vaccine to prevent herpes zoster and post-herpetic neuralgia in older adults. PMID- 16263053 TI - Chronic meningitis. AB - Chronic meningitis is a syndrome commonly defined by the presence of continuously persistent meningeal inflammation for at least 4 weeks. Presenting neurologic features are often nonspecific, and the list of differential diagnoses is broad. Despite the development of modern molecular diagnostic methods, establishing a specific cause may challenge the acumen of the treating neurologist with clues to the diagnosis residing outside the nervous system and requiring focused investigation. This review discusses selected etiologies illustrating issues in diagnosis and etiologic categories to be considered in the evaluation of this challenging syndrome. PMID- 16263056 TI - Mild cognitive impairment: to treat or not to treat. AB - Mild cognitive impairment (MCI) refers to memory deficits in excess of normal aging, but not sufficient for the diagnosis of Alzheimer's disease (AD). In general, patients with MCI are not disabled and have no other obvious cognitive deficits other than memory. Many studies have shown that patients with MCI are more likely to progress to AD than age-matched controls. Drug therapy to prevent or delay deterioration in patients with MCI has been tested only very recently. The results of three clinical trials are discussed. One of two trials with donepizil suggests that donepizil may delay onset of AD, but the effect is modest and the side effects are problematic. Galantamine appeared to be associated with excess deaths, and vitamin E was not effective. No medications are currently indicated for the prevention of dementia in patients with MCI. PMID- 16263054 TI - Post-infectious encephalomyelitis. AB - The term post-infectious encephalomyelitis (PIEM) is frequently used interchangeably with acute disseminated encephalomyelitis (ADEM), although technically PIEM occurs after a known infection whereas with ADEM there is no antecedent infection. PIEM represents one of the primary demyelinating disorders of the central nervous system, along with multiple sclerosis and Devic's disease. There is no specific diagnostic test for any of these conditions and at onset it may be difficult to differentiate between ADEM and the first attack of multiple sclerosis. However, there are clinical and magnetic resonance imaging features that allow differentiation between PIEM/ADEM and a relapsing disease such as multiple sclerosis. Some patients improve spontaneously; most improve with methylprednisolone. If that fails, plasma exchange or intravenous immunoglobulin may be effective. PMID- 16263055 TI - Central nervous system Lyme disease. AB - Nervous system infection with Borrelia burgdorferi frequently causes meningitis and rarely causes encephalomyelitis. Altered cognitive function also can occur in the absence of central nervous system infection. Recently developed serodiagnostic tools, such as the C6 assay, and appropriate use of Western blotting promise to improve diagnostic accuracy. Treatment trials have demonstrated the efficacy of relatively brief courses of oral antimicrobial agents, even in peripheral nervous system infection and meningitis. Several well performed studies have clearly shown that prolonged antimicrobial treatment of "post-Lyme disease" is ineffective. Diagnosis and treatment of Lyme disease continue to improve. PMID- 16263058 TI - Robotics and other devices in the treatment of patients recovering from stroke. AB - Stroke is the leading cause of permanent disability in the United States despite advances in prevention and novel interventional treatments. Randomized controlled studies have demonstrated the effectiveness of specialized post-stroke rehabilitation units, but administrative orders have severely limited the length of stay, so novel approaches to the treatment of recovery need to be tested in outpatients. Although the mechanisms of stroke recovery depend on multiple factors, a number of techniques that concentrate on enhanced exercise of the paralyzed limb have demonstrated effectiveness in reducing the motor impairment. For example, interactive robotic devices are new tools for therapists to deliver enhanced sensorimotor training for the paralyzed upper limb, which can potentially improve patient outcome and increase patient productivity. New data support the idea that for some post-stroke patients and for some aspects of training-induced recovery, timing of the training may be less important than the quality and intensity of the training. The positive outcome that resulted in the interactive robotic trials contrasts with the failure to find a beneficial result in trials that used a noninteractive device that delivered continuous passive motion only. New pilot data from novel devices to move the wrist demonstrate benefit and suggest that successive improvement of the function of the arm progressing to the distal muscles may eventually lead to significant disability reduction. These data from robotic trials continue to contribute to the emerging scientific basis of neuro-rehabilitation. PMID- 16263059 TI - The STICH trial: the end of surgical intervention for supratentorial intracerebral hemorrhage? PMID- 16263057 TI - Aphasia and right hemisphere syndromes in stroke. AB - This article highlights the latest findings regarding the effect of acute and chronic stroke on behavior, specifically left hemispheric injury causing aphasia and right hemispheric injury causing neglect, visual-spatial problems, and other cognitive syndromes. We review papers published in the past 2 years pertaining to localization, assessment, recovery, treatment, and outcomes of aphasia and right hemisphere cognitive syndromes following stroke. PMID- 16263060 TI - Update on respiratory management of critically ill neurologic patients. AB - Respiratory failure complicates a number of acute neurologic conditions, most notably neuromuscular diseases (eg, Guillain-Barre syndrome and myasthenia gravis) and stroke. In addition, pulmonary complications, particularly pneumonia and atelectasis, are fairly common in patients with these diagnoses; their prevention and early recognition are crucial to avoid detrimental consequences. This review discusses recent studies related to predictors of respiratory failure and pneumonia, strategies of respiratory care and ventilatory support, functional prognosis, and withdrawal of mechanical ventilation in patients with acute neuromuscular respiratory failure and stroke. PMID- 16263061 TI - Recent advances in the management of acute intracerebral hemorrhage. AB - Intracerebral hemorrhage (ICH) is the most deadly form of stroke, and its acute treatment has suffered from a lack of guidance by reliable clinical trial data. In the past year, however, important clinical trials have helped point toward effective acute management. Studies have shown that magnetic resonance imaging is as accurate as computed tomography in diagnosing acute ICH, although this study is not always feasible in critically ill patients. Ultra-early hemostatic therapy has shown promise in limiting early hematoma expansion and rebleeding. The role of early surgery in patient management has been partially clarified. Finally, a novel treatment for intraventricular hemorrhage has shown promise in speeding clot resolution. All of these advances provide grounds for optimism that multimodal, evidence-based treatment of acute ICH will be reality in the near future. PMID- 16263062 TI - Cardiopulmonary complications of brain injury. AB - Cardiac and pulmonary complications following acute neurologic injury are common and may be a cause of morbidity and mortality in this population. Examples include hypertension, arrhythmias, ventricular dysfunction, pulmonary edema, shock, and sudden death. Primary neurologic events are represented by stroke, subarachnoid hemorrhage, traumatic brain injury, epilepsy, and encephalitis and have been frequently reported. Given the high frequency of these conditions, it is important for physicians to become familiar with their pathophysiology, allowing for more prompt and appropriate treatment. PMID- 16263064 TI - Effects of lamb age, hormone stimulation and response to hormone stimulation on the yield and in vitro developmental competence of prepubertal lamb oocytes. AB - Experiments were conducted to determine the effects of lamb age, hormone stimulation (Experiment 1) and response to stimulation (Experiment 2) on the in vitro production of embryos from prepubertal lambs aged 3-4 and 6-7 weeks of age. For 3-4-week-old lambs, hormone stimulation increased the number of follicles (29.9 +/- 15.3 v. 70.6 +/- 8.2), oocytes per ovary (18.3 +/- 6.3 v. 39.3 +/- 5.8) and oocyte development to the blastocyst stage (0/192 (0.0%) v. 115/661 (17.4%); P < 0.05). Lamb age (3-4 v. 6-7 weeks old) increased oocyte development to the blastocyst stage (115/661 (17.4%) v. 120/562 (21.4%) respectively). In Experiment 2, hormone-stimulated lambs (3-4 and 6-7 weeks old) were divided into low, medium or high responders based on the number of ovarian follicles (<20, 20-50 and >100 follicles per ovary respectively). The response to hormone stimulation did not affect oocyte recovery rate, but the number of oocytes suitable for culture was increased for high-responding 3-4-week-old lambs only (P < 0.05). Oocyte development to the blastocyst stage was not affected by response to stimulation for 3-4-week-old lambs (15.2-25.6%; P > 0.05), but was reduced for high (6.7%) compared with low (19.5%) and medium (30.9%) responding 6-7-week-old lambs (P < 0.05). These results demonstrate that the production of embryos from prepubertal lambs is increased by hormone stimulation and lamb age and the response to stimulation does not affect embryo production from 3-4-week-old lambs, although by 6-7 weeks of age a high response to stimulation reduces blastocyst formation. PMID- 16263063 TI - Treatment of massive cerebral infarction. AB - Stroke is the third leading cause of death in the United States, with a person dying every 3 minutes of a stroke. Massive ischemic stroke accounts for 10% to 20% of ischemic strokes, has traditionally been associated with a high mortality and morbidity, and requires a unique management strategy. Recent advances in management, fueled by an increased understanding of the pathophysiology, may help decrease mortality and improve outcomes. Rapid access to reperfusion therapies remains the most critical element of stroke care and the cornerstone of therapy. This article focuses on newer therapies, including osmotic therapy, hypothermia, maintained normothermia, strict glycemic control, induced hypertension, and hemicraniectomy, all of which show promise for reducing mortality and improving functional outcome. These interventions have become integrated into neurologic intensive care units around the world. They are complicated, require a high level of expertise, and carry a significant learning curve. In order for these new management techniques to be effective, an expedited, aggressive, meticulous, and potentially prolonged medical management approach is needed. To accomplish this there is a growing need for focused specialists in the areas of neurointensive care and stroke. PMID- 16263066 TI - Comparative study of sperm chromatin condensation in the excurrent ducts of the laboratory mouse Mus musculus and spinifex hopping mouse Notomys alexis. AB - In most mammals, post-testicular sperm maturation is completed in the caput and corpus epididymides, with storage occurring in the cauda epididymides. However, in the spinifex hopping mouse, Notomys alexis, epididymal sperm transit is rapid and some sperm storage occurs in the distal region of the vas deferens. The aim of the present study was to determine whether the rapid progression of sperm into the vas deferens in the hopping mouse results in late sperm maturation. To determine this, sperm nuclei from the epididymides and vasa deferentia of laboratory and hopping mice were compared for: (1) thiol content after staining with monobromobimane (mBBr); (2) chromatin resistance to acid denaturation following incubation with acetic alcohol and staining with acridine orange; and (3) chromatin resistance to in vitro decondensation after incubation with 1% sodium dodecyl sulfate (SDS). It was found that, whereas laboratory mouse sperm completed chromatin condensation by the time they reached the cauda epididymidis, hopping mouse sperm nuclei from the vas deferens showed significantly less mBBr fluorescence and a greater proportion of sperm were resistant to decondensation with SDS than those in the cauda epididymidis. Therefore, the results of the present study indicate that, unlike in the laboratory mouse, hopping mouse chromatin condensation of spermatozoa continues in the vas deferens and this may be due, at least in part, to rapid epididymal transit. PMID- 16263065 TI - Ontogeny and pathway of formation of 5alpha-androstane-3alpha,17beta-diol in the testes of the immature brushtail possum Trichosurus vulpecula. AB - The testicular androgen 5alpha;-androstane-3alpha,17beta-diol (androstanediol) mediates virilisation in pouch young of a marsupial, the tammar wallaby, and is the principal androgen formed in immature rodent testes. To chart the pattern of androstanediol formation in another marsupial species, the testes or fragments of testes from brushtail possums (Trichosurus vulpecula) that spanned the age range from early pouch young to mature adults were incubated with (3)H-progesterone and the products were identified by high-performance liquid chromatography. The only 19-carbon steroids identified in pouch young and adult testes were the Delta(4)-3 keto-steroids testosterone and androstenedione. However, androstanediol and another 5alpha-reduced androgen (androsterone) were synthesised by testes from Day 87-200 males and these appeared to be formed from the 5alpha-reduction and 3 keto reduction of testosterone and androstenedione. In the prostate and glans penis of the immature male, (3)H-androstanediol was converted to dihydrotestosterone. We conclude that the timing of androstanediol formation in the possum testis resembles the process in rodents rather than in the tammar wallaby and that any androstanediol in the circulation probably acts in target tissues via conversion to dihydrotestosterone. PMID- 16263067 TI - Deficit in reproduction in polygynously mated females of the monogamous mound building mouse Mus spicilegus. AB - In monogamous mound-building mice Mus spicilegus, facultative polygyny was observed in April, at the beginning of the reproductive season. In order to evaluate the cost of polygyny, we compared, under laboratory conditions, the reproductive success of trios and pairs composed of sister females mated with unrelated brother males. Females in trios were able to tolerate each other during the reproductive period and to reproduce with the same male under spatial constraints. Nevertheless, polygyny had a strong negative effect on the reproductive success of the females. The average number of young per litter was smaller in trio females than in paired females, whereas the interval between two successive litters was higher. As a result, the number of litters and the number of young per time unit were smaller in each of the trio females compared with paired females. Agonistic behaviour being absent in trios, our results strongly suggest that living in trios led to pheromonal production that affected the female physiological state and reproduction. Males of trios did not obtain a better reproductive success than males in pairs. PMID- 16263068 TI - Identification of metaphase II-specific gene transcripts in porcine oocytes and their expression in early stage embryos. AB - Annealing control primer (ACP)-based GeneFishing polymerase chain reaction (PCR) was used to identify the genes that are specifically or prominently expressed in porcine oocytes at the metaphase II (MII) and germinal vesicle (GV) stages. By using 60 ACPs, 13 differentially expressed genes (DEGs) were identified. The cloned genes or expressed sequence tags (ESTs) showed sequence similarity with known genes or ESTs of other species in GenBank. The mRNA expression during oocyte maturation and early embryonic development in both pigs and mice of four of these genes (namely transcription factor TZP, annexin A2, hypoxia-inducible protein 2, and ATPase 6) was further characterised by real-time quantitative reverse transcription-PCR. All four genes were markedly upregulated in pig and mouse MII oocytes compared with GV-stage oocytes. The expression levels of the four genes decreased gradually during early cleavage. Thus, these genes may play important roles during oocyte maturation and/or early cleavage in mammals. Although the detailed functions of these genes remain to be determined, their identification in the present study provides insights into meiotic maturation and fertilisation. PMID- 16263069 TI - Inositol transport in mouse embryonic stem cells. AB - The uptake of myo-inositol by mouse embryonic stem (ES) cells was measured using [2-(3)H]myo-inositol. Uptake of myo-inositol by ES cells occurred in a mainly saturable, sodium-, time- and temperature-dependent manner, which was inhibited by glucose, phloridzin and ouabain. Self inhibition by inositol was much greater than inhibition by glucose indicating that transport was not occurring via a sodium-dependent glucose transporter. Uptake rate was much greater than efflux rate indicating a mainly unidirectional transport mechanism. Estimated kinetics parameters for sodium-dependent inositol uptake were a K m of 65.1 +/- 11.8 micromol L(-1) and a V max of 5.0 +/- 0.59 pmol microg protein(-1) h(-1). Inositol uptake was also sensitive to osmolality; uptake increased in response to incubation in hypertonic medium indicating a possible role for inositol as an osmolyte in ES cells. These characteristics indicate that myo-inositol transport in mouse ES cells occurs by a sodium-dependent myo-inositol transporter protein. PMID- 16263071 TI - Effect of homologous follicular fluid from medium-sized and large follicles on in vitro maturation of equine cumulus-oocyte complexes. AB - The in vitro maturation (IVM) of equine oocytes is typically performed using various synthetic media; however, an optimal IVM system for equine oocytes has not been developed. The aim of the present study was to evaluate the effects of two types of follicular fluid (FF) obtained from cyclic mares and two incubation intervals for the IVM of equine cumulus-oocyte complexes (COCs). Follicular fluid was collected from medium-sized (20-29 mm diameter) and large (e30 mm; post-human chorionic gonadotrophin administration) follicles using transvaginal ultrasound guided follicle aspiration. Compact (n = 232) and non-compact (n = 183) COCs obtained from a slaughterhouse were incubated separately in the following groups: (1) FF from medium follicles for 24 h; (2) FF from large follicles for 24 h; (3) control (synthetic) medium for 24 h; (4) FF from medium follicles for 24 h then FF from large follicles for an additional 24 h; (5) FF from large follicles for 48 h; and (6) control medium for 48 h. For compact COCs, there was a tendency (P = 0.06) for more COCs incubated in FF from large follicles for 24 h to reach metaphase II compared with those incubated in control medium for 24 h (58% v. 35%, respectively). More (P < 0.05) compact COCs had degenerated after incubation in control medium for 48 h compared with all other groups (51% v. 14-24%, respectively). For non-compact COCs, incubation in FF from medium follicles for 24 h resulted in more (P = 0.05) COCs at metaphase II compared with control medium for 48 h (58% v. 29%, respectively). These results indicate that homologous FF from cyclic mares is a suitable alternative for the IVM of equine COCs and that it may be superior to conventional media for longer (i.e. >24 h) incubation intervals. PMID- 16263070 TI - Fluoxetine during pregnancy: impact on fetal development. AB - Women are at greatest risk of suffering from depression during the childbearing years and thus may either become pregnant while taking an antidepressant or may require a prescription for one during pregnancy. The antidepressant fluoxetine (FX) is a selective serotonin reuptake inhibitor (SSRI), which increases serotonin neurotransmission. Serotonin is involved in the regulation of a variety of physiological systems, including the sleep-wake cycle, circadian rhythms and the hypothalamic-pituitary-adrenal axis. Each of these systems also plays an important role in fetal development. Compared with other antidepressant drugs, the SSRIs, such as FX, have fewer side effects. Because of this, they are now frequently prescribed, especially during pregnancy. Clinical studies suggest poor neonatal outcome after exposure to FX in utero. Recent studies in the sheep fetus describe the physiological effects of in utero exposure to FX with an 8 day infusion during late gestation in the sheep. This is a useful model for determining the effects of FX on fetal physiology. The fetus can be studied for weeks in its normal intrauterine environment with serial sampling of blood, thus permitting detailed studies of drug disposition in both mother and fetus combined with monitoring of fetal behavioural state and cardiovascular function. Fluoxetine causes an acute increase in plasma serotonin levels, leading to a transient reduction in uterine blood flow. This, in turn, reduces the delivery of oxygen and nutrients to the fetus, thereby presenting a mechanism for reducing growth and/or eliciting preterm delivery. Moreover, because FX crosses the placenta, the fetus is exposed directly to FX, as well as to the effects of the drug on the mother. Fluoxetine increases high-voltage/non-rapid eye movement behavioural state in the fetus after both acute and chronic exposure and, thus, may interfere with normal fetal neurodevelopment. Fluoxetine also alters hypothalamic function in the adult and increases the magnitude of the prepartum rise in fetal cortisol concentrations in sheep. Fetal FX exposure does not alter fetal circadian rhythms in melatonin or prolactin. Studies of the effects of FX exposure on fetal development in the sheep are important in defining possible physiological mechanisms that explain human clinical studies of birth outcomes after FX exposure. To date, there have been insufficient longer-term follow-up studies in any precocial species of offspring exposed to SSRIs in utero. Thus, further investigation of the long-term consequences of in utero exposure to FX and other SSRIs, as well as the mechanisms involved, are required for a complete understanding of the impact of these agents on development. This should involve studies in both humans and appropriate animal models. PMID- 16263072 TI - Progesterone receptors and proliferating cell nuclear antigen expression in equine luteal tissue. AB - Steroid hormones act via specific receptors, and these play an important physiological role in the ovary. The objective of this study was to evaluate the cellular distribution of progesterone receptors and their staining intensity in different equine luteal structures during the breeding season, as well as their relationship to luteal cell composition, cell proliferation pattern and plasma progesterone (P4) concentration. There was an increase in proliferating cell nuclear antigen (PCNA) expression in large luteal cells from the corpus hemorrhagicum (CH) to mid-luteal phase, followed by a decrease toward the late luteal stage. In the CH, the number of large luteal cells was lower than in other structures. Only large luteal cells showed positive staining for P(4) receptors. An increase in staining intensity for P(4) receptors was observed between CH and mid-phase corpus luteum, and CH and late-phase corpus luteum. Synthesis of P(4) started at a very early stage of the luteal structure and was accompanied by an increase in P(4) receptors and PCNA expression, and proliferation of large luteal cells, until mid-luteal phase. These data suggest that large luteal cells might play an important role in the regulation or synthesis of P(4) in equine luteal structures. PMID- 16263073 TI - Safe pedestrian crossings for children and elderly. AB - In May 2000, the Swedish code governing the conduct of drivers at marked crosswalks became stricter with the intent to improve safety and mobility for pedestrians. A crash analysis based on a macro study of all of Sweden suggests that the injury risk in marked, not reconstructed, crosswalks increased by 27% for pedestrians and 19% for bicyclists. The reason for this may be that pedestrians get a false sense of safety with the new code. Reconstructions aiming at lowering speeds are indeed needed for the change of code to be positive. The 90-percentile speed should not exceed 30 km/h or safety will deteriorate. However, low speed by itself may not guarantee optimal safety. Safety can be further improved at sites, which already have been reconstructed to ensure low speeds. Results based on field data collected at sites close to schools in Malmo, Trollhattan and Boras in Sweden, and in-depth studies and other analyses of Finnish and Swedish police-reported crashes, suggest that safety of children and elderly is further improved at sites where visibility, orientation and clarity are sufficient. Also, marking crosswalks may increase yield rates (expected improvement 6%) towards pedestrians; and speed cushions situated at a longer distance from the marked crosswalk increase yield rates towards pedestrians and cyclists compared to speed cushions closer by. PMID- 16263074 TI - Leprosy and confinement due to leprosy show high association with hepatitis C in Southern Brazil. AB - Leprosy is a disease, which is accompanied by cellular immunity defects, which may increase the susceptibility of patients in developing co-infections. The association of leprosy with hepatitis C virus (HCV) infection, human immunodeficiency virus types 1 and 2 (HIV 1+2) infection and human T-lymphotropic virus types I and II (HTLV I+II) infection have previously been described in different populations. In this study, the prevalence of these infections was determined in 199 Southern Brazilian leprosy patients and in 681 matched controls. Antibodies to HCV were positive in 3.52% of the patients (7/199) and in 0.15% of the controls (1/681; odds ratio (OR)=24.79; 95% CI=3.03-202.74; p=0.0002). An increased risk of HCV infection was observed in institutionalized patients (OR=14.95; 95% CI=1.76-127.03; p=0.004) and in the lepromatous form of the disease (OR=7.67; 95% CI=0.43-136.62; p=ns). Anti-HIV 1+2 antibodies were positive in only one out-patient (1/199; 0.50%) and in none of the controls (0/681; OR=3.43; 95% CI=0.21-55.16; p>0.05). No leprosy patient was positive for anti-HTLV I+II antibodies. These results demonstrate an increased prevalence of HCV infection in leprosy patients from South Brazil and that both institutionalization and lepromatous form of the disease confer higher risk to HCV infection. These data emphasizes the importance of monitoring hepatitis C and leprosy interactions and the need of special care to institutionalized and lepromatous patients in preventing HCV co-infection. PMID- 16263075 TI - Alterations of temporalis muscle contractile force and histological content from the myostatin and Mdx deficient mouse. AB - Myostatin (GDF8) and dystrophin are critical molecules for muscle organisation. Myostatin is involved in regulating muscle growth and development, whereas dystrophin is part of the dystrophin-glycoprotein complex (DGC), which anchors the cytoskeleton to the sarcolemma. We examined temporalis muscle morphology and function in myostatin deficient and dystrophin deficient (Mdx) mice in order to determine how myostatin and dystrophin affect bite force and muscle fibre composition. Bite forces from 4-month-old myostatin-/-, dystrophin deficient (Mdx) and normal control mice were measured by load cell and field stimulation of the temporalis muscle. Tissue sections were stained with haemotoxylin and eosin (H&E) and the periodic acid-Schiff reaction (PAS) to assess morphology and fibre type differences. A positive relationship between bite force and muscle mass for both genetic models was observed. Both Mstn-/- and Mdx mice showed significant elevation in bite force and muscle mass. Histological examination revealed greater muscle fibre cross-sectional area variability in Mdx mice (ANOVA, F=5.6, P<0.01). Surprisingly, the Mstn-/- mice demonstrated a disproportionate increase in bite force at higher stimulation frequencies with comparison of regression lines for force-frequency data (ANOVA, F=3.46, P<0.07). Muscle fibre typing using a PAS staining technique revealed significantly more type IIx/b glycolytic muscle fibres in the Mstn-/- mice. Our results suggest that histopathologies associated with Mdx mice did not diminish gross temporalis structure or function, whilst the force-frequency changes associated with Mstn-/- mice were reflected in an elevation of type IIx/b fibres. PMID- 16263076 TI - Connexins as targets for cancer chemoprevention and chemotherapy. AB - Cells within a tissue continuously interact to coordinate normal tissue functions and maintain homeostasis. Gap junctional communication (GJC), mediated by the connexin protein family, allows this type of intercellular crosstalk resulting in synchronized and cooperative tissue behavior such as cardiac contraction. In cancer, loss of these types of cell:cell interactions has been shown to facilitate tumorigenesis and enable the autonomous cell behavior associated with transformed cells. Indeed, many human tumor lines demonstrate deficient or aberrant GJC and/or loss of connexin expression. Restoration of exogenous connexin expression/GJC function is correlated with increased cell growth control both in vitro and in vivo. In support of this growth regulatory hypothesis, decreased connexin expression has been observed in situ in early human neoplasia of various organs. Additionally, genetically engineered mice lacking particular connexins (Connexins 32 or 43) exhibit increased susceptibility to radiation and chemically-induced liver and/or lung tumorigenesis. These studies strongly suggest that connexins and GJC serve a tumor suppressor role. Consistent with this proposed role, in a model cell culture system, retinoids and carotenoids up regulate Connexin43 (Cx43) expression in direct proportion to their ability to suppress carcinogen-induced neoplastic transformation. Here, we discuss the important role of connexins and GJC in tumorigenesis and suggest the possibility of connexins as potential anti-oncogenic targets for chemoprevention and/or chemotherapy. PMID- 16263077 TI - Structural insights into the function of human caveolin 1. AB - Caveolin-1 (Cav-1) is emerging as the central protein controlling caveolae formation, caveolae trafficking, and cellular signalling. In particular, it is known that Cav-1 interacts and modulates the activity of several signalling proteins through the so-called caveolin scaffolding domain. In this paper, we used a bioinformatics approach to assess the validity of some long-standing structural features of Cav-1. We could confirm the existence of a membrane spanning region of Cav-1 and highlight an interesting pattern of palmitoylated cysteine residues explaining the structural features of the Cav-1 C-terminal region. Moreover, the scaffolding domain is predicted to have a different structure than previously reported. PMID- 16263078 TI - Cross-reactivity of antibody against SARS-coronavirus nucleocapsid protein with IL-11. AB - Infection of SARS-associated coronavirus (SARS-CoV) induced a strong anti nucleocapsid (anti-N) antibody response. However, the pathophysiological significance of the anti-N antibodies in SARS pathogenesis is largely unknown. To profile the anti-N antibodies, a phage-displayed scFv library was prepared from mice immunized with heat-inactivated SARS-CoV-infected Vero E6 cell lysate. Specific anti-N scFvs were isolated by panning against a recombinant nucleocapsid protein and reactivity was confirmed with phage-ELISA. Sequence analysis indicated that two of the isolated anti-N scFv clones were identical and displayed a high homology with an scFv specific for interleukin 11 (IL-11), an anti-inflammatory cytokine derived from bone marrow stroma cells. In a neutralization assay, IL-11-induced STAT 3 phosphorylation in rat intestinal epithelial IEC-18 cells was completely suppressed by the anti-N scFv clone L9N01. PMID- 16263079 TI - Predicting alternate structure attainment and amyloidogenesis: a nonlinear signal analysis approach. AB - Chain hydrophobicity values have been used in prediction of alternate structure attainment by a polypeptide. Nonlinear signal analysis on the hydrophobicity values gives important clues about the propensities of particular stretches of a protein to form local or nonlocal contacts. These contacts determine the folding behavior of a polypeptide and helps in predicting the final structure that can be attained. A nonlinear signal analysis called the recurrent quantification analysis has been carried out using the hydrophobicity values on a wide range of proteins obtained from human, plant, and fungal sources. Here, we show that such an analysis gives us an easy handle in determining sequences within the proteins that may be important in beta-sheet formation leading to amyloidosis. PMID- 16263080 TI - DNA damage and repair in mammalian cells exposed to p-hydroxyphenylpyruvic acid. AB - Tyrosinemia type 1 (HT1) is an autosomal recessive disorder of the tyrosine metabolism in which the fumarylacetoacetate hydrolase enzyme is defective. This disease is clinically heterogeneous and a chronic and acute form is discerned. Characteristic of the chronic form is the development of cellular hepatocarcinoma. Although p-hydroxyphenylpyruvic acid (pHPPA) is used as one of the diagnostic markers of this disease, it was suggested that it is unlikely to be involved in the pathophysiology of HT1 as it is present in other disorders that does not have hepatorenal symptoms. It was the aim of this study to investigate the possible effect of pHPPA on DNA damage and repair in mammalian cells. The comet assay was used to establish the genotoxicity of pHPPA in human peripheral blood lymphocytes and isolated rat hepatocytes after their exposure to pHPPA. At first glance the damage to DNA caused by pHPPA seemed reparable in both cell types, however, after challenging the DNA repair capacity of metabolite treated cells with treatment with H(2)O(2), a marked impairment in the DNA repair capability of these cells was observed. We suggest that the main effect of pHPPA is the long-term impairment of the DNA repair machinery rather than the direct damage to DNA and that this effect of pHPPA, together with the other characteristic metabolites, e.g., FAA and MAA, causes cellular hepatocarcinoma to develop in the chronic form of HT1. PMID- 16263081 TI - Activation of (Na+ + K+)-ATPase. AB - Enzymes catalyze essential chemical reactions needed for living processes. (Na+ +K+)-ATPase (NKA) is one of the key enzymes that control intracellular ion homeostasis and regulate cardiac function. Little is known about activation of NKA and its biological impact. Here we show that native activity of NKA is markedly elevated when protein-protein interaction occurs at the extracellular DVEDSYGQQWTYEQR (D-R) region in the alpha-subunit of the enzyme. The apparent catalytic turnover of NKA is approximately twice as fast as the controls for both ouabain-resistant and ouabain-sensitive enzymes. Activation of NKA not only markedly protects enzyme function against denaturing, but also directly affects cellular activities by regulating intracellular Ca2+ transients and inducing a positive inotropic effect in isolated rat cardiac myocytes. Immunofluorescent labeling indicates that the D-R region of NKA is not a conventional digitalis binding site. Our findings uncover a novel activation site of NKA that is capable of promoting the catalytic function of the enzyme and establish a new concept that activating of NKA mediates cardiac contraction. PMID- 16263082 TI - Direct measurement of the rupture force of single pair of decorin interactions. AB - Decorin is one important member of the family of small leucine-rich proteoglycans, which are widely distributed in connective tissues in the body such as tendon and ligament. Decorin may be responsible for collagen fibril connection in those tissues. A recent hypothesis suggests that decorin may bind to collagen with its core protein while binding to another decorin through the interaction with their glycosaminoglycan (GAG) chains. However, there is no direct evidence supporting this hypothesis to date. In this study, the interaction of decorin GAG chains was directly determined for the first time. The rupture force of single bonds between decorins (GAG chains interaction) was determined directly as 16.5+/-5.1 pN using a laser tweezers/interferometer single molecular nanomechanical testing system. This information can improve our understanding of the mechanical properties of connective tissues at the molecular level. PMID- 16263083 TI - Depressed glutathione synthesis precedes oxidative stress and atherogenesis in Apo-E(-/-) mice. AB - Glutathione is a vital intracellular antioxidant. The enzymes involved in its synthesis and utilisation are tightly regulated, but the importance of glutathione regulation in atherogenesis is poorly understood. Here, we establish that glutathione is severely (approximately 80%) depleted very early (10 weeks) in the atheroma-prone aortic arch of male apoprotein E-deficient (Apo-E(-/-)) mice compared to age-matched wild-type controls. Importantly, this event pre empts lipid peroxidation and detectable atheroma by several months. Depletion of glutathione was associated with excessive oxidant burden and reduced transcription and activity of the rate-limiting enzyme for glutathione synthesis, gamma-glutamylcysteine ligase, together with the glutathione-dependent antioxidant enzyme, glutathione peroxidase. Depletion via reduced synthesis of glutathione precedes lipid peroxidation and atherogenesis in Apo-E(-/-) mice. We suggest that glutathione deficiency is central to the failure of the intracellular antioxidant defences and is causally implicated in the pathogenesis of atherosclerosis. Modification of the glutathione pathway may present a novel and important therapeutic target in the prevention and treatment of atherosclerosis. PMID- 16263084 TI - Zcchc8 is a glycogen synthase kinase-3 substrate that interacts with RNA-binding proteins. AB - Phosphorylation of c-Myc on threonine 58 (T58) stimulates its degradation by the Fbw7-SCF ubiquitin ligase. We used a phosphorylation-specific antibody raised against the c-Myc T58 region to attempt to identify other proteins regulated by the Fbw7 pathway. We identified two predominant proteins recognized by this antibody. The first is Ebna1 binding protein 2, a nucleolar protein that, in contrast with a previous report, is likely responsible for the nucleolar staining exhibited by this antibody. The second is Zcchc8, a nuclear protein that is highly phosphorylated in cells treated with nocodazole. We show that Zcchc8 is directly phosphorylated by GSK-3 in vitro and that GSK-3 inhibition prevents Zcchc8 phosphorylation in vivo. Moreover, we found that Zcchc8 interacts with proteins involved in RNA processing/degradation. We suggest that Zcchc8 is a GSK 3 substrate with a role in RNA metabolism. PMID- 16263085 TI - Membrane-anchored CD14 is required for LPS-induced TLR4 endocytosis in TLR4/MD 2/CD14 overexpressing CHO cells. AB - Lipopolysaccharide (LPS) induces inflammatory activation through TLR4 (toll-like receptor-4)/MD-2 (myeloid differentiation-2)/CD14 (cluster of differentiation-14) complex. Although optimal LPS signaling is required to activate our innate immune systems against gram-negative bacterium, excessive amount of LPS signaling develops a detrimental inflammatory response in gram-negative bacterial infections. Downregulation of surface TLR4 expression is one of the critical mechanisms that can restrict LPS signaling. Here, we found that membrane-anchored CD14 is required for LPS-induced downregulation of TLR4 and MD-2 in CHO cells. Moreover, pretreatment of the cells with sterol-binding agent filipin reduced LPS induced TLR4 downregulation, suggesting the involvement of caveolae-mediated endocytosis pathway. Involvement of caveolae in LPS-induced TLR4 endocytosis was further confirmed by immunoprecipitation. Thus, our data indicate that caveolae dependent endocytosis pathway is involved in LPS-induced TLR4 downregulation and that this is dependent on membrane-anchored CD14 expression. PMID- 16263086 TI - Selective enhancement of tonic inhibition by increasing ambient GABA is insufficient to suppress excitotoxicity in hippocampal neurons. AB - Gamma-aminobutyric acid (GABA) activates synaptic GABA(A) receptors to generate inhibitory postsynaptic potentials. GABA also acts on extrasynaptic GABA(A) receptors, resulting in tonic inhibition. The physiological role of tonic inhibition, however, remains elusive. We explored the neurophysiological significance of tonic inhibition by testing whether selective activation of extrasynaptic GABA(A) receptors is sufficient to curb excitotoxicity. Tonic inhibition was selectively enhanced by increasing ambient GABA. In both acute hippocampal slices and cultured hippocampal neurons, boosting tonic inhibition alone is insufficient to withstand the hyper-excitability of hippocampal neurons induced by low-magnesium (Mg2+) baths. Furthermore, selective activation of extrasynaptic GABA(A) receptors resulted in no significant neuroprotective effects against glutamate or low-Mg2+-induced neuronal cell deaths. These data imply that under physiological conditions extrasynaptic GABA(A) receptors are optimally activated by ambient GABA and that a further increase in extracellular GABA concentration will not significantly enhance the effect of tonic inhibition on neuronal excitability. PMID- 16263087 TI - The protein phosphatase-1 targeting subunit TIMAP regulates LAMR1 phosphorylation. AB - TIMAP is a prenylated endothelial cell protein with a domain structure that predicts it to be a protein phosphatase-1 (PP-1) regulatory subunit. We found that TIMAP interacts with the 37/67 kDa laminin receptor (LAMR1) in yeast two hybrid assays. In endothelial cells, endogenous TIMAP and LAMR1 co immunoprecipitated and co-localized at the plasma membrane. TIMAP amino acids 261 290, representing the fourth ankyrin repeat of TIMAP, are necessary and sufficient for the interaction. In MDCK cells, lacking endogenous TIMAP, overexpression of full-length TIMAP, but not TIMAP deleted in the fourth ankyrin domain, allowed co-immunoprecipitation with LAMR1. PP-1 co-precipitated with overexpressed and endogenous TIMAP in MDCK and endothelial cells, respectively. In MDCK cells, PP-1 associated with LAMR1 in the presence, but not in the absence, of TIMAP. LAMR1 was a substrate for PP-1 in vitro, and in MDCK cells its phosphorylation was abrogated by expression of full-length TIMAP but not by TIMAP deficient in the fourth ankyrin domain. Hence, TIMAP targets PP-1 to LAMR1, and LAMR1 is a TIMAP-dependent PP-1 substrate. PMID- 16263089 TI - Detection of kappa and delta opioid receptors in skin--outside the nervous system. AB - Opioid receptors (OR) are widely expressed in the central nervous system (CNS). Opioid antinociception might be initiated by activation of OR outside the CNS, indicating targeting of peripheral OR could be useful in the treatment of chronic pain. This study was designed to detect OR in skin tissues of healthy volunteers at both mRNA and protein levels. Skin samples from 10 healthy individuals were investigated. Total isolated RNAs were reverse transcribed, amplified and quantified by real-time PCR. Tissue and skin fibroblast OR protein was detected by immunohistochemistry, Western blot, and immunofluorescence. All skin tissue samples expressed delta- (DOR) and kappa-OR (KOR) mRNAs. Using immunohistochemistry, DOR and KOR were localized in skin fibroblast-like and mononuclear cells. Skin fibroblasts in culture expressed DOR and KOR mRNA. Using immunofluorescence, both DOR and KOR proteins were expressed predominantly on the cell membrane with minor staining in the cytoplasm. We suggest that enhanced expression of DOR and KOR in skin justifies the exploration of selective novel delta and kappa agonists for local pain treatment. PMID- 16263088 TI - Cell-free synthesis of recombinant proteins from PCR-amplified genes at a comparable productivity to that of plasmid-based reactions. AB - The functional stability of mRNA is one of the crucial factors affecting the efficiency of cell-free protein synthesis. The importance of the stability of mRNA in the prolonged synthesis of protein molecules becomes even greater when the cell-free protein synthesis is directed by PCR-amplified DNAs, because the linear DNAs are rapidly degraded by the endogenous nucleases and, thus, the continuous generation of mRNA molecules is limited. With the aim of developing a highly efficient cell-free protein synthesis system directed by PCR products, in this study, we describe a systematic approach to enhance the stability of mRNA in cell-free extracts. First, exonuclease-mediated degradation was substantially reduced by introducing a stem-loop structure at the 3'-end of the mRNA. The endonucleolytic cleavage of the mRNA was minimized by using an S30 extract prepared from an Escherichia coli strain that is deficient in a major endonuclease (RNase E). Taken together, through the retardation of the endonucleolytic and exonucleolytic degradations of the mRNA molecules, the level of protein expression from the PCR-amplified DNA templates becomes comparable to that of conventional plasmid-based reactions. The enhanced productivity of the PCR-based cell-free protein synthesis enables the high-throughput generation of protein molecules required for many post-genomic applications. PMID- 16263090 TI - Fibroblast growth factor 3, a protein with a dual subcellular fate, is interacting with human ribosomal protein S2. AB - The secreted isoform of fibroblast growth factor 3 (FGF3) induces a mitogenic cell response, while the nuclear form inhibits cell proliferation. Recently, we identified a nucleolar FGF3-binding protein which is implicated in processing of pre-rRNA as a possible target of nuclear FGF3 signalling. Here, we report a second candidate protein identified by a yeast two-hybrid screen for nuclear FGF3 action, ribosomal protein S2, rpS2. Recombinant rpS2 binds to in vitro translated FGF3 and to nuclear FGF3 extracted from transfected COS-1 cells. Characterization of the FGF3 binding domain of rpS2 showed that both the Arg-Gly-rich N-terminal region and a short carboxyl-terminal sequence of rpS2 are necessary for FGF3 binding. Mapping the S2 binding domains of FGF3 revealed that these domains are important for both NoBP and rpS2 interaction. Transient co-expression of rpS2 and nuclear FGF3 resulted in a reduced nucleolar localization of the FGF. These findings suggest that the nuclear form of FGF3 inhibits cell proliferation by interfering with ribosomal biogenesis. PMID- 16263091 TI - Drug specificity and intestinal membrane localization of human organic cation transporters (OCT). AB - This study was performed to investigate which human organic cation transporter, hOCT1, hOCT2 or hOCT3, participates with regard to cation specificity and membrane localization in the intestinal absorption of orally available cationic drugs. Inhibition of N-[methyl-3H]4-phenylpyridinium ([3H]MPP+) uptake by various compounds into Caco-2 cells and into cells (HEK-293 or CHO) that were stably transfected with hOCT1, hOCT2 or hOCT3 was compared. The uptake of [3H]MPP+ into Caco-2 cells was inhibited by atropine, butylscopolamine, clonidine, diphenhydramine, etilefrine, quinine and ranitidine with IC50 values between 6 microM and 4 mM. Transepithelial, apical to basal flux of [3H]MPP+ across Caco-2 cell monolayers was also strongly inhibited by these compounds. The inhibitory potency of the cationic drugs and prototypical organic cations at Caco-2 cells correlated well with the inhibitory potency measured at CHO-hOCT3 cells but much less with that at HEK-hOCT1 and -hOCT2 cells. This is functional evidence for the predominant role of hOCT3. Etilefrine and atropine were specifically transported into CHO cells by hOCT3. In Caco-2 cells, the mRNA of all three hOCT and the proteins hOCT2 and hOCT3 were detected. More importantly, immunocytochemical analyses of human jejunum revealed for the first time that hOCT3 is localized to the brush border membrane whereas hOCT1 immunolabeling was mainly observed at the lateral membranes of the enterocytes. PMID- 16263092 TI - Inhibition of sphingolipid synthesis impairs cellular activation, cytokine production and proliferation in human lymphocytes. AB - The localisation of the T cell receptor and other signalling molecules in membrane microdomains (MM) is essential for the activation of T lymphocytes. These MM are stabilized by sphingolipids and cholesterol. It was recently shown that the activation of T lymphocytes leads to the confluence of small MM and the formation of an immunological synapse which is thought to be essential for a persistent activation and proliferation. We studied the effects of an inhibition of sphingolipid synthesis on T lymphocyte function. Both sphingolipid inhibitors, PDMP and myriocin, inhibited glucosphingolipids in whole cell lipid extracts and in MM. Both compounds inhibited the proliferation of superantigen-stimulated PBMC without inducing cell death. However, only the ceramide-like compound PDMP inhibited the expression of activation markers and the secretion of IFN-gamma which was not seen with myriocin treatment. The MM localisation of Lck and LAT was not significantly reduced in PDMP-treated cells. In conclusion, our results show that glucosphingolipids are necessary for cell growth of human T lymphocytes. However, inhibition of glucosphingolipid synthesis itself did not inhibit cellular activation. Our data show that glucosphingolipids - in contrast to cholesterol - are not essential for the stabilisation of MM. PMID- 16263093 TI - Increased lysosomal uptake of methotrexate-polyglutamates in two methotrexate resistant cell lines with distinct mechanisms of resistance. AB - Methotrexate (MTX) resistance in mitoxantrone-selected MCF7/MX cells and in MTX selected CEM/MTX cells is associated with reduced drug accumulation, albeit caused by different mechanisms. In addition, in both resistant cell lines the proportion of active long-chain MTX-polyglutamate (MTX-PG) metabolites is reduced relative to that in the respective parental cell line. Previous studies by others have implied that increased lysosomal uptake could affect the rate of MTX-PG hydrolysis, and hence the length distribution of the polyglutamate chains. However, in the two cell line pairs studied, the number of lysosomes per cell was not different between the corresponding parental and resistant cells. Instead, we observed a two- to three-fold increased facilitative uptake of MTX-Glu4 by the lysosomes from these two independently derived MTX-resistant cell lines, compared to uptake by lysosomes from their corresponding parental cells. Enhanced lysosomal uptake of MTX-Glu4 was reflected in an increased maximal uptake velocity, without a change in the apparent substrate affinity. In addition, the rate of MTX efflux from lysosomes from CEM/MTX cells was two-fold faster than from lysosomes from CEM cells. Consistent with this observation, the relative amount of short-chain MTX-Glu(1+2) species, as a fraction of the total amount of all MTX-Glu(1-4) species combined, was only half as large in lysosomes from CEM/MTX cells as in lysosomes from CEM cells. Together, these results suggest the possibility that increased lysosomal uptake, and hence enhanced sequestration of MTX-PGs in resistant cells, contributes to the development of high-level MTX resistance by decreasing the cytosolic levels of MTX-PGs. PMID- 16263095 TI - Normal hypocretin-1 (orexin-A) levels in the cerebrospinal fluid of patients with Huntington's disease. AB - A significant atrophy and loss of hypocretin neurons in the brains of human patients with Huntington's disease (HD) and in R6/2 mice have been reported. We included 10 patients with HD and 12 patients with chorea-like hyperkinetic movement disorders (non-HD). All patients of the HD group and eleven patients of the non-HD group showed normal hypocretin-1 levels. Thus, hypocretin-1 may not serve as an additional diagnostic marker for HD. PMID- 16263094 TI - The effect of 'binge' cocaine administration on the expression of cyclin dependent kinase 5 and its activator p35 in various regions of rat brain. AB - The present study was aimed at determining whether the administration of cocaine in 'binge' pattern regimen that evoked tolerance to the locomotor stimulant effects of cocaine also influenced the expression of cyclin-dependent kinase 5 (Cdk5) and its activator p35 in the amygdala, medial prefrontal cortex, nucleus accumbens septi and caudate-putamen. Western blot techniques revealed that acute and repeated 'binge' cocaine decreased expression of the Cdk5 protein in the amygdala. In the medial prefrontal cortex, only exposure to repeated 'binge' cocaine decreased the content of the Cdk5 protein. 'Binge' cocaine administration also altered the expression of Cdk5 activator p35 protein. In the amygdala, only repeated 'binge' cocaine decreased the expression of p35, while in the medial prefrontal cortex, a decrease was observed after acute and repeated 'binge' cocaine exposure. In neither the nucleus accumbens septi nor the caudate-putamen acute or repeated 'binge' cocaine modified the expression of Cdk5 and p35. The above data indicate that in contrast to sensitizing doses of cocaine, a single and repeated binge of cocaine, which evoked tolerance to its locomotor stimulant effects, decreases expression of Cdk5 and p35 and possibly decreases the efficacy of neurotransmission or induces brain plastic changes regulated by Cdk5 and its activator p35. PMID- 16263096 TI - Tracer coupling of neurons in the rat retina inner nuclear layer labeled by Fluorogold. AB - A subpopulation of neurons in the inner nuclear layer (INL) of the rat retina were labeled 9-13 weeks after application of Fluorogold (FG) to the superior colliculus. Neurobiotin injection of FG-labeled cells in the INL of flatmounted living retina revealed that these cells consisted of both displaced ganglion cells and a subset of amacrine cells. Fluorogold-labeled amacrine cells in the INL showed tracer coupling to other presumptive amacrine cells in the INL, but there was no evidence of coupling to neurons in the ganglion cell layer (GCL). As the labeling of amacrine cells by FG may be due to gap junction coupling between ganglion and amacrine cells, these data add to the evidence that tracer coupling between these cells can be unidirectional. Some of the FG-labeled displaced ganglion cells in the INL injected with Neurobiotin also showed tracer coupling to neurons in the INL or GCL. PMID- 16263097 TI - Trace amines reduce GABA(B) receptor-mediated presynaptic inhibition at GABAergic synapses of the rat substantia nigra pars compacta. AB - Trace amines (TAs) act in the mammalian brain through amphetamine-like effects and as endogenous agonists of specific receptors. We now show that tyramine and beta-phenylethylamine, in the presence of specific dopamine (DA) receptor antagonists, inhibit the GABA(B)-dependent presynaptic inhibition of GABAergic inputs to midbrain DA neurons. Our results further extend the role of TAs as neuromodulators and propose a novel mechanism by which they modulate DA neurons. PMID- 16263098 TI - The nitrosteroid NCX 1015, a prednisolone derivative, improves recovery of function in rats after spinal cord injury. AB - Glucocorticoids, given at high-doses, improve recovery of function after spinal cord injury (SCI) in animals. However, side effects combined with a limited efficacy in clinical trials have restricted their usefulness for treatment of SCI patients. Recent studies have shown that incorporation of the nitric oxide releasing moiety into the glucocorticoid structure enhances anti-inflammatory properties and reduces side effects. One compound, a derivative of prednisolone (PRE), (NCX 1015, prednisolone 21 [(4'nitrooxymethyl)benzoate]), has interesting pharmacological properties. Therefore, we investigated its effects on apoptosis and recovery of function in rats after SCI. Rats received subcutaneously vehicle, NCX 1015 or PRE (37 micromol/kg, each) 3.5 h after a standardized thoracic lesion. The treatment was continued once a day for 3 days and the effect of both steroids on apoptosis was examined by immunohistochemistry 24 h after the last injection. NCX 1015 but not PRE reduced TUNEL and activated caspase 3 in both white and ventral gray matter as well as tumor necrosis factor immunoreactivity in ventral horn motorneurons, suggesting that NCX 1015 reduces SCI-induced apoptosis. The effect of NCX 1015 on motor function was then examined by a standard locomotion rating scale (BBB) starting at 1 day after injury and continuing up to 14 days. NCX 1015 improved significantly locomotor activity by 4 days after injury, whereas PRE had an effect equivalent to that of vehicle, thus providing a correlation between the antiapoptotic effect of NCX1015 and its ability to improve recovery of function. The data suggest that NCX 1015 might be a novel experimental therapeutic compound for recovery of function in SCI patients. PMID- 16263099 TI - The majority of bladder sensory afferents to the rat lumbosacral spinal cord are both IB4- and CGRP-positive. AB - The rat urinary bladder is innervated by neurons in dorsal root ganglia (DRG) that express the neuropeptides calcitonin gene-related peptide (CGRP) and substance P (SP), and a fraction of bladder afferents can bind the non peptidergic marker isolectin B4 (IB4). We used histochemical binding and axonal tracing to identify the bladder afferents, and immunocytochemistry to determine the degree of colocalization of CGRP with IB4 in their cell bodies in DRG and in their central axons in the spinal cord. In the L6 DRG, about 60% of CGRP-positive neurons were also positive for IB4. In the spinal cord, IB4 and CGRP colocalized in fibers and terminals in the inner part of lamina II, the lateral collateral path, and the sacral parasympathetic nucleus (SPN). In SPN, the majority of IB4 positive fibers and terminals were also CGRP-positive. After injection of IB4 into the bladder wall, immunoreaction for IB4 was detected in SPN, but not in lamina II. These results suggest that most IB4-positive afferents from the bladder are also CGRP-positive, and that the distinction between peptidergic and non-peptidergic bladder afferents based on IB4 binding is of limited validity. PMID- 16263100 TI - Respiratory and autonomic responses to microinjection of NMDA and AMPA into the commissural subnucleus of the NTS of awake rats. AB - The changes in mean arterial pressure (MAP) and respiratory frequency (RF) in response to microinjection of NMDA or AMPA into the commissural subnucleus of the NTS (comNTS) at the calamus scriptorius level of awake rats were evaluated. Under tribromoethanol anesthesia, the rats received guide-cannulae in direction of the NTS and a catheter was inserted into the femoral artery for measurement of arterial pressure. Changes in RF were evaluated with the rats inside a plethysmographic chamber. Randomly microinjections of 5 doses of NMDA (0.001, 0.01, 0.1, 1.0 and 2 nmol/50 nL; n = 10) or AMPA (1, 5, 10, 25 and 50 pmol/50 nL; n = 8) into the comNTS were performed at 15 min intervals and produced a dose dependent increase in MAP [NMDA (3 +/- 2, 4 +/- 3, 25 +/- 4, 41 +/- 4 and 51 +/- 4 mm Hg) and AMPA (0 +/- 1, 14 +/- 4, 17 +/- 3, 27 +/- 5 and 34 +/- 3 mm Hg)]. Microinjection of NMDA (1 nmol/50 nL; n = 7) or AMPA (50 pmol/50 nL; n = 4) into the comNTS produced a long lasting apnea. The pressor responses to microinjection of NMDA or AMPA into the comNTS were blocked by prazosin, a alpha(1)-adrenoceptor antagonist, indicating that the increase in arterial pressure in both cases was sympathetically mediated. The data show that microinjection of NMDA and AMPA into the comNTS produced pressor response and apnea, indicating that both ionotropic l glutamate receptors may play a role in the neurotransmission/neuromodulation of the autonomic and respiratory components of the cardiovascular reflexes at this level. PMID- 16263101 TI - A short synthesis of D-glycero-D-manno-heptose 7-phosphate. AB - D-glycero-D-manno-Heptopyranose 7-phosphate-an intermediate in the biosynthesis of nucleotide-activated heptoses-has been prepared in good overall yield from benzyl 5,6-dideoxy-2,3-O-isopropylidene-alpha-D-lyxo-(Z)-hept-5-enofuranoside by a short-step synthesis. Phosphitylation using the phosphoramidite procedure followed by in situ oxidation afforded the corresponding 7-O-phosphotriester derivative in high yield. Subsequent osmylation proceeded in good diastereoselectivity (4:1) to furnish the D-glycero-D-manno-configured derivative, which was separated from the L-glycero-L-gulo-isomer by chromatography. Hydrogenolysis led to simultaneous removal of the benzyl and isopropylidene groups and afforded the target compound in high yield, which serves as a substrate of bacterial heptose 7-phosphate kinases. PMID- 16263102 TI - Synthesis, structure and reactivity of 5-pyranosyl-1,3,4-oxathiazol-2-ones. AB - 5-(1,2,3,4-tetra-O-acetyl-alpha-D-xylopyranos-5S-C-yl)-1,3,4-oxathiazol-2-one (8) has been prepared from glucuronamide in two steps and 73% overall yield by conversion to the tetra-O-acetyl derivative 7 followed by reaction with chlorocarbonylsulfenyl chloride. 5-(2,3,4-Tri-O-acetyl-beta-D-xylopyranosyl) 1,3,4-oxathiazol-2-one (12) was synthesised from D-xylose by a four-step sequence involving conversion to the xylopyranosylnitromethane derivative 9, reaction with PCl3 to afford nitrile 10, hydrolysis to amide 11, and finally treatment with ClCOSCl. D-glucose-derived analogue 13 was prepared similarly. The structure of oxathiazolone 8 was established by X-ray crystallography. Thermolysis of the oxathiazolones 8 and 12 at 130-160 degrees C resulted in decarboxylation and desulfuration to yield the corresponding nitriles. Attempts to trap the putative nitrile sulfide intermediates by repeating the thermolysis in the presence of dipolarophiles, such as ethyl cyanoformate, afforded only traces of the 1,3 dipolar cycloadducts; however, under microwave irradiation oxathiazolone 8 and ethyl cyanoformate afforded ethyl 3-(1,2,3,4-tetra-O-acetyl-alpha-D-xylopyranos 5S-C-yl)-1,2,4-thiadiazole-5-carboxylate 22 in good yield. PMID- 16263103 TI - Correlation between red blood cell acetylcholinesterase activity and neuromuscular transmission in organophosphate poisoning. AB - Assessment of effectiveness of oximes in severely organophosphate poisoned patients is hampered by sedation, artificial ventilation and other therapeutic measures as well as varying individual clinical courses due to, e.g. differences in type and amount of poison ingested or time elapsed before treatment starts. To evaluate oxime effects a suitable surrogate parameter would be helpful. Red blood cell acetylcholinesterase (RBC-AChE) is easily obtainable, shows a similar structure as synaptic enzyme and may be useful to reflect the AChE status at the synaptic site. Accordingly, it appeared rational to check whether RBC-AChE activity could be correlated with neuromuscular transmission (NMT), an easily accessible clinical parameter. The correlation was assessed in a clinical trial with severely OP-poisoned patients who were treated with obidoxime. The investigation revealed a good correlation between both parameters and showed, that a very low RBC-AChE activity (<10% of normal) was associated with a strongly impaired NMT marker, the so called decrement-phenomenon, RBC-AChE activity between 10 and 30% by impaired NMT with the decrement-increment-phenomenon and RBC activities above 30% generally by normal muscle function. Accordingly, RBC AChE appears to be a suitable parameter for judgment of oxime effectiveness at the neuromuscular junction, one of the most important targets for therapy where atropine is ineffective in OP-poisoning. PMID- 16263104 TI - Human serum butyrylcholinesterase: in vitro and in vivo stability, pharmacokinetics, and safety in mice. AB - The use of exogenously administered cholinesterases (ChEs) as bioscavengers of highly toxic organophosphate (OP) nerve agents is now sufficiently well documented to make them a highly viable prophylactic treatment against this potential threat. Of the ChEs evaluated so far, human serum butyrylcholinesterase (HuBChE) is most suitable for human use. A dose of 200 mg (3 mg/kg) of HuBChE is envisioned as a prophylactic treatment in humans that can protect from an exposure of up to 2 x LD50 of soman. In addition to its use as a prophylactic for a variety of wartime scenarios, including covert actions, it also has potential use for first responders (civilians) reacting to terrorist nerve gas release. We recently, developed a procedure for the large-scale purification of HuBChE, which yielded approximately 6 g of highly purified enzyme from 120 kg of Cohn fraction IV-4. The enzyme had a specific activity of 700-750 U/mg and migrated as a single band on SDS-PAGE. To provide data for initiating an investigational new drug (IND) application for the use of this enzyme as a bioscavenger in humans, we established its pharmacokinetic properties, examined its safety in mice, and evaluated its shelf life at various temperatures. In mice administered various doses up to 90 mg/kg, enzyme activity reached peak levels in circulation at 10 and 24 h following i.p. and i.m. injections, respectively. The enzyme displayed a mean residence time (MRT) of 40-50 h, regardless of the route of administration or dose of injected enzyme. Mice were euthanized 2 weeks following enzyme administration and tissues were examined grossly or microscopically for possible toxic effects. Results suggest that HuBChE does not exhibit any toxicity in mice as measured by general observation, serum chemistry, hematology, gross or histologic tissue changes. The shelf life of this enzyme stored at 4, 25, 37, and 45 degrees C was determined in lyophilized form. The enzyme was found to be stable when stored in lyophilized form at -20, 4, 25, or 37 degrees C to date (2 years), as measured by specific activity and SDS polyacrylamide gel electrophoresis. The effect of storage on circulatory stability was determined by measuring MRT in mice; there was no change in the MRT of lyophilized enzyme stored at -20 degrees C to date (2 years). These results provide convincing data that HuBChE is a safe bioscavenger that can provide protection against all OP nerve agents. Efforts are now underway to prepare the required documentation for submission of an IND application to the United States Food and Drug Administration (USFDA). PMID- 16263105 TI - A numerical coupling model to analyze the blood flow, temperature, and oxygen transport in human breast tumor under laser irradiation. AB - The aim of this study is to investigate the variation of the blood perfusion rate and distribution of oxygen partial pressure (PO2) in human tumors by a coupling numerical model when laser irradiation is used as an adjuvant method in the treatment of cancer. A two-dimensional finite element (FE) thermal model of a human breast with a tumor was developed. The blood circulation inside the breast was modeled using one-dimensional non-linear equations of pulsatile fluid flow. The distribution of PO2 inside the capillaries, tumor vessels, and surrounding tissue was obtained by the Krogh analysis model. Finally, the variations of the average tumor temperature, blood perfusion, and PO2 during laser heating were computed by coupling the blood circulation, FE thermal, and oxygen transport models. PMID- 16263106 TI - Reconstruction and representation of caudal vasculature of zebrafish embryo from confocal scanning laser fluorescence microscopic images. AB - Three-dimensional (3D) reconstruction from a series of sections is an important technique in medical imaging, particularly for visualization of blood vessels from angiography. Here, we present a framework for automatic segmentation and registration of different kind of blood vessels from 2-day-old zebrafish embryos. Series of optical sections were acquired from confocal microscopy with the blood vessels labeled by fluorescent microbeads (0.02 microm) injected into blood stream of 2-day-old zebrafish embryos. Blood vessels were extracted and their morphological parameters, including length and diameter, were calculated. At the same time, individual blood vessels were registered automatically. Vasculature was represented by attributed vessel represent graph (AVRG), which contained morphological data and connectivity of every blood vessel. Using AVRG to represent a vasculature made the comparison between vasculatures of different embryos more easy. Visualization, as well as quantification, of reconstructed 3D model of AVRG was presented in an interactive interface. The framework was implemented by Visual C++ as Windows-based program. PMID- 16263108 TI - Imbalance between drug and non-drug reward availability: a major risk factor for addiction. AB - Laboratory animals self-administer most, though not all, drugs of abuse. Recent evidence shows that with increased drug availability, most laboratory rats develop all the major behavioral signs of addiction, including: 1) drug intake escalation, 2) increased motivation for the drug, 3) difficulty to abstain, 4) decreased reward function, and 5) inflexible drug use. The large prevalence of addicted rats may suggest that they are particularly vulnerable to develop compulsive drug use. I review evidence showing that this apparent vulnerability results in large part from the lack of positive (i.e., alternative non-drug rewards) and negative (i.e., costs) incentives capable of turning animals away from the pursuit of drugs. In particular, most animals seem to take drugs and eventually become addicted, not because drugs are intrinsically addictive, but more likely because drugs are the only significant sources of reward available in the laboratory. Laboratory animals would therefore represent more of a model of high-risk human groups than of the general population. Consequently, they should be more suited for searching factors that protect from, rather than predispose to, drug addiction. Reconsidering the environmental background of drug self administration experiments in laboratory animals raises intriguing implications for understanding the initial demand for drug consumption and the transition to drug addiction, and for extrapolation from laboratory animals to humans. PMID- 16263107 TI - Corneal keratocytes retain neural crest progenitor cell properties. AB - Corneal keratocytes have a remarkable ability to heal the cornea throughout life. Given their developmental origin from the cranial neural crest, we asked whether this regenerative ability was related to the stem cell-like properties of their neural crest precursors. To this end, we challenged corneal stromal keratocytes by injecting them into a new environment along cranial neural crest migratory pathways. The results show that injected stromal keratocytes change their phenotype, proliferate and migrate ventrally adjacent to host neural crest cells. They then contribute to the corneal endothelial and stromal layers, the musculature of the eye, mandibular process, blood vessels and cardiac cushion tissue of the host. However, they fail to form neurons in cranial ganglia or branchial arch cartilage, illustrating that they are at least partially restricted progenitors rather than stem cells. The data show that, even at late embryonic stages, corneal keratocytes are not terminally differentiated, but maintain plasticity and multipotentiality, contributing to non-neuronal cranial neural crest derivatives. PMID- 16263109 TI - Do similar neural systems subserve aggressive and sexual behaviour in male rats? Insights from c-Fos and pharmacological studies. AB - It is a common belief that male aggressive and sexual behaviour share many of the underlying neurobiological, neurological, pharmacological and neuroendocrine mechanisms. Therefore, we studied brain activation patterns in male rat after performance of aggressive and sexual behaviour and compared serotonergic pharmacology in the same paradigms to delineate possible similarities and differences. Patterns of Fos-immunoreactivity induced by aggressive and sexual encounters of Wild-type male Brown Norway rats were studied to localise the commonly activated (functionally shared) parts of the circuitry, and the specific (functionally different) parts of the neuronal circuitry. Some brain areas (caudal medial preoptic area and medial amygdala) were commonly activated, but other areas (e.g. posterodorsal parts of the medial amygdala, rostral preoptic and premammillary hypothalamus) showed remarkably specific differences in neural activation. 5-HT(1A) receptor agonists inhibit aggressive, but stimulate male sexual behaviour, whereas 5-HT(1B) receptor agonists inhibit both types of behaviour. Selective serotonin reuptake inhibitors share comparable inhibitory effects in aggression and sexual behaviour, although only at relatively high doses. We propose that separate hard-wired neural systems exist in the brain for aggressive and sexual behaviours, modulated via hierarchically 'higher-level' brain areas that are involved in the integration (gating) of the behavioural outcome of an organism. PMID- 16263110 TI - C-type natriuretic peptide applied to the brain enhances exocrine pancreatic secretion through a vagal pathway. AB - C-type natriuretic peptide (CNP) is the major natriuretic peptide in the brain and its mRNA has been reported in the central nervous system, which supports local synthesis and its role as a neuromodulator. The aim of the present work was to study the effect of centrally applied CNP on pancreatic secretion. Rats were fitted with a lateral cerebroventricular cannula one-week before secretion studies. The central administration of CNP dose-dependently enhanced pancreatic fluid and protein output. CNP response was diminished by atropine and hexamethonium, but it was abolished by vagotomy. Neither adrenergic antagonists nor the administration of (D-p-Cl-Phe(6),Leu(17))-vasoactive intestinal peptide (VIP antagonist) or N(omega) Nitro-L arginine methyl ester (L-NAME) (nitric oxide synthase inhibitor) affected CNP response. The effect induced by CNP was mimicked by 8-Br-cGMP but not by c-ANP-(4-23) amide (selective agonist of the natriuretic peptide receptor C). Furthermore, CNP interacted with cholecystokinin (CCK) and secretin in the brain to modify pancreatic secretion. Present findings show that centrally applied CNP enhanced pancreatic secretion through a vagal pathway and suggest that CNP response is mediated by the activation of natriuretic peptide guanylyl cyclase coupled receptors in the brain. PMID- 16263112 TI - Phenotypic heterogeneity influences the behavior of rat aortic smooth muscle cells in collagen lattice. AB - Phenotypic modulation of vascular smooth muscle cells (SMCs) in atherosclerosis and restenosis involves responses to the surrounding microenvironment. SMCs obtained by enzymatic digestion from tunica media of newborn, young adult (YA) and old rats and from the thickened intima (TI) and underlying media of young adult rat aortas 15 days after ballooning were entrapped in floating populated collagen lattice (PCL). TI-SMCs elongated but were poor at PCL contraction and remodeling and expressed less alpha2 integrin compared to other SMCs that appeared more dendritic. During early phases of PCL contraction, SMCs showed a marked decrease in the expression of alpha-smooth muscle actin and myosin. SMCs other than TI-SMCs required 7 days to re-express alpha-smooth muscle actin and myosin. Only TI-SMCs in PCL were able to divide in 48 h, with a greater proportion in S and G2-M cell cycle phases compared to other SMCs. Anti-alpha2 integrin antibody markedly inhibited contraction but not proliferation in YA-SMC PLCs; anti-alpha1 and anti-alpha2 integrin antibodies induced a similar slight inhibition in TI-SMC-PCLs. Finally, TI-SMCs rapidly migrated from PCL on plastic reacquiring their epithelioid phenotype. Heterogeneity in proliferation and cytoskeleton as well the capacity to remodel the extracellular matrix are maintained, when SMCs are suspended in PCLs. PMID- 16263111 TI - Integration of topographical and biochemical cues by axons during growth on microfabricated 3-D substrates. AB - During embryonic neural development, axon tips ("growth cones") are guided through a dynamic three-dimensional (3-D) landscape by soluble chemotropic factors and by immobilized, growth-permissive or growth-inhibiting contact cues present in the extracellular matrix and on the surface of surrounding cells. It has been difficult to probe the search algorithms of growth cones in response to multiple contact cues during 3-D navigation using traditional two-dimensional (2 D) substrates. Here, we present an in vitro study in which the axons of murine embryonic cortical neurons are challenged with competing growth options, using 3 D substrates that feature variations in permissiveness and microtopography. As 3 D substrates, we used poly-D-lysine (PDL) coatings on microfabricated steps of polydimethylsiloxane (PDMS) and complementary features of Matrigel. We found that axons display a preference for PDL over Matrigel and for the straightest path within a distance consistent with the exploratory range of the growth cone. When these two preferences are in conflict, axons choose to grow straight into Matrigel; when the straight path is not permissive, the axon turns in the direction that minimizes the turning angle. These results suggest that growth cones make 3-D navigation decisions by integrating permissiveness and topographical cues. PMID- 16263113 TI - Optical biosensor provides insights for bradykinin B(2) receptor signaling in A431 cells. AB - The spatial and temporal targeting of proteins or protein assemblies to appropriate sites is crucial to regulate the specificity and efficiency of protein-protein interactions, thus dictating the timing and intensity of cell signaling and responses. The resultant dynamic mass redistribution could be manifested by label free optical biosensor, and lead to a novel and functional optical signature for studying cell signaling. Here we applied this technology, termed as mass redistribution cell assay technology (MRCAT), to study the signaling networks of bradykinin B(2) receptor in A431 cells. Using MRCAT, the spatial and temporal relocation of proteins and protein assemblies mediated by bradykinin was quantitatively monitored in microplate format and in live cells. The saturability to bradykinin, together with the specific and dose-dependent inhibition by a B(2) specific antagonist HOE140, suggested that the optical signature is a direct result of B(2) receptor activation. The sensitivity of the optical signature to cholesterol depletion by methyl-beta-cyclodextrin argued that B(2) receptor signaling is dependent on the integrity of lipid rafts; disruption of these microdomains hinders the B(2) signaling. Modulations of several important intracellular targets with specific inhibitors suggested that B(2) receptor activation results in signaling via at least dual pathways - G(s)- and G(q)-mediated signaling. Remarkably, the two signaling pathways counter regulate each other. Several critical downstream targets including protein kinase C, protein kinase A, and epidermal growth factor receptor had been identified to involve in B(2) signaling. The roles of endocytosis and cytoskeleton modulation in B(2) signaling were also demonstrated. PMID- 16263114 TI - The N-terminal cleavage of cellular prion protein in the human brain. AB - Human brain cellular prion protein (PrP(c)) is cleaved within its highly conserved domain at amino acid 110/111/112. This cleavage generates a highly stable C-terminal fragment (C1). We examined the relative abundance of holo- and truncated PrP(c) in human cerebral cortex and we found important inter-individual variations in the proportion of C1. Neither age nor postmortem interval explain the large variability observed in C1 amount. Interestingly, our results show that high levels of C1 are associated with the presence of the active ADAM 10 suggesting this zinc metalloprotease as a candidate for the cleavage of PrP(c) in the human brain. PMID- 16263115 TI - Epigenetic and classical activation of Entamoeba histolytica heat shock protein 100 (EHsp100) expression. AB - The protozoan parasite Entamoeba histolytica expresses a cytosine-5 DNA methyltransferase (Ehmeth) that belongs to the DNMT2 protein family. The biological function of members of this DNMT2 family is unknown. In the present study, the 5' region of E. histolytica heat shock protein 100 (5'EHsp100) was isolated by affinity chromatography with 5-methylcytosine antibodies as ligand. The methylation status of 5'EHsp100 was confirmed by sodium bisulfite sequencing. We showed that the expression of EHsp100 was induced by heat shock, 5-azacytidine (5-AzaC), an inhibitor of DNA methyltransferase and Trichostatin A (TSA), an inhibitor of histone deacetylase. The effect of TSA on EHsp100 expression was rapidly reversed by removing the drug from the culture. In contrast, EHsp100 expression was still detectable one month after removing 5-AzaC from the media. Whereas 5-AzaC and TSA caused demethylation in the promoter region of EHsp100, no demethylation was observed following heat shock. Remarkably, DNA that includes three putative heat shock elements identified in the promoter region of EHsp100 bound to a protein of 37kDa present in the nuclear fraction of heat-shocked trophozoites but absent in the nuclear fraction of 5-AzaC and TSA treated trophozoites. Our data suggest that EHsp100 expression can be regulated by both a classical and an epigenetic mechanism. PMID- 16263116 TI - Plasma membrane and lysosomal localization of CB1 cannabinoid receptor are dependent on lipid rafts and regulated by anandamide in human breast cancer cells. AB - In this report we show, by confocal analysis of indirect immunofluorescence, that the type-1 cannabinoid receptor (CB1R), which belongs to the family of G-protein coupled receptors, is expressed on the plasma membrane in human breast cancer MDA MB-231 cells. However, a substantial proportion of the receptor is present in lysosomes. We found that CB1R is associated with cholesterol- and sphyngolipid enriched membrane domains (rafts). Cholesterol depletion by methyl-beta cyclodextrin (MCD) treatment strongly reduces the flotation of the protein on the raft-fractions (DRM) of sucrose density gradients suggesting that CB1 raft association is cholesterol dependent. Interestingly binding of the agonist, anandamide (AEA) also impairs DRM-association of the receptor suggesting that the membrane distribution of the receptor is dependent on rafts and is possibly regulated by the agonist binding. Indeed MCD completely blocked the clustering of CB1R at the plasma membrane. On the contrary the lysosomal localization of CB1R was impaired by this treatment only after AEA binding. PMID- 16263117 TI - The PDZ/coiled-coil domain containing protein PIST modulates insulin secretion in MIN6 insulinoma cells by interacting with somatostatin receptor subtype 5. AB - The multi-domain protein PIST (protein interacting specifically with Tc10) interacts with the SSTR5 (somatostatin receptor 5) and is responsible for its intracellular localization. Here, we show that PIST is expressed in pancreatic beta-cells and interacts with SSTR5 in these cells. PIST expression in MIN6 insulinoma cells is reduced by somatostatin (SST). After stimulation with SST, SSTR5 undergoes internalization together with PIST. MIN6 cells over-expressing PIST display enhanced glucose-stimulated insulin secretion and a decreased sensitivity to SST-induced inhibition of insulin secretion. These data suggest that PIST plays an important role in insulin secretion by regulating SSTR5 availability at the plasma membrane. PMID- 16263118 TI - Sensitivity of the Drosophila para sodium channel to DDT is not lowered by the super-kdr mutation M918T on the IIS4-S5 linker that profoundly reduces sensitivity to permethrin and deltamethrin. AB - DDT inhibits Na channel inactivation and deactivation, promotes Na channel activation and reduces the resting potential of Xenopus oocytes expressing the Drosophila para Na channel. These changes are only marginally influenced by the single mutation M918T (super-kdr) but are reduced approximately 10-fold by either the single mutation L1014F (kdr) or the double mutation L1014F+M918T, both of which confer resistance to the pyrethroids permethrin and deltamethrin. We conclude that DDT binds either to or in the region of L1014 on IIS6 but only weakly to M918 on the IIS4-S5 linker, which is part of a high-affinity binding site for permethrin and deltamethrin. PMID- 16263119 TI - Structural characterization of the [Pco/o(2)] compound of cytochrome c oxidase. AB - The structural properties of a key transient oxygen intermediate of cytochrome c oxidase, P(R), remain an enigma, although inferences have been drawn from its equilibrium analogues, [Pco/o(2)] , P(H) and P(M). With resonance Raman spectroscopy, an oxygen isotope-sensitive band at 806 cm(-1) was observed in [Pco/o(2)] produced by adding CO and O(2) to the resting enzyme. The vibrational band shifted to 771 cm(-1) upon isotopic substitution of (16)O(2) with (18)O(2). The same modes at 806 and 771 cm(-1) were present simultaneously when the mixed isotope, (18)O(16)O, was employed, indicating that in [Pco/o(2)] the O-O bond is cleaved, resulting in a Fe(4+)O(2-) structure. This result unifies the nature of the three equilibrium analogues of the P(R) intermediate. PMID- 16263120 TI - Regulation of EGF receptor signaling by the MARVEL domain-containing protein CKLFSF8. AB - It is known that chemokine-like factor superfamily 8 (CKLFSF8), a member of the CKLF superfamily, has four putative transmembrane regions and a MARVEL domain. Its structure is similar to TM4SF11 (plasmolipin) and widely distributed in normal tissue. However, its function is not yet known. We show here that CKLFSF8 is associated with the epidermal growth factor receptor (EGFR) and that ectopic expression of CKLFSF8 in several cell lines suppresses EGF-induced cell proliferation, whereas knockdown of CKLFSF8 by siRNA promotes cell proliferation. In cells overexpressing CKLFSF8, the initial activation of EGFR was not affected, but subsequent desensitization of EGF-induced signaling occurred rapidly. This attenuation was correlated with an increased rate of receptor endocytosis. In contrast, knockdown of CKLFSF8 by siCKLFSF8 delayed EGFR endocytosis. These results identify CKLFSF8 as a novel regulator of EGF-induced signaling and indicate that the association of EGFR with four transmembrane proteins is critical for EGFR desensitization. PMID- 16263121 TI - A proteomic snapshot of the human heat shock protein 90 interactome. AB - Heat shock protein 90 (Hsp90) is a molecular chaperone which modulates several signalling pathways within a cell. By applying co-immunoprecipitation with endogeneous Hsp90, we were able to identify 39 novel protein interaction partners of this chaperone in human embryonic kidney cells (HEK293). Interestingly, levels of DNA-activated protein kinase catalytic subunit, an Hsp90 interaction partner found in this study, were found to be sensitive to Hsp90 inhibitor treatment only in HeLa cells but not in HEK293 cells referring to the tumorgenicity of this chaperone. PMID- 16263122 TI - CD45 expression on rat acinar cells: involvement in pro-inflammatory cytokine production. AB - CD45 transduces activation signals in inflammatory cells. We investigate CD45 expression on pancreatic acinar cells and examine its role in the inflammatory response which these cells have also shown under certain circumstances. Similar CD45 mRNA levels were found in acinar cells and leukocytes (positive control). Flow cytometric and immunohistochemical analysis showed a heterogeneous CD45 distribution on acinar cells. Activation of acinar cells by incubation with pancreatitis-associated ascitic fluid as evidencied by TNF-alpha production resulted in a decreased CD45 expression, suggesting that CD45 acts as a negative regulator of cytokine production. As a validation of this finding in vivo, a decrease in the acinar CD45 expression in parallel with an increased ability to produce TNF-alpha was found in rats with acute pancreatitis. Our data show that CD45 is constitutively expressed in acinar cells and suggest that it plays an important role in negatively regulating cytokine production. PMID- 16263123 TI - Analysis of expression of growth factor receptors in replicatively and oxidatively senescent human fibroblasts. AB - Replicatively and oxidatively senescent human fibroblasts demonstrate an impaired response to mitogens. To investigate whether this is due to downregulation of growth factor receptors we examined their expression in these two types of senescence. mRNA and protein levels of the insulin receptor and platelet-derived growth factor (PDGF) alpha-receptor decreased in replicatively senescent cells. The PDGF beta-receptor and insulin-like growth factor 1 receptor at the protein level also decreased but remained readily detectable. However, these major growth factor receptors remained unchanged in oxidatively premature senescent cells. This suggests that mechanisms underlying diminished responsiveness to mitogens might be different in replicative senescence and oxidatively premature senescence. PMID- 16263124 TI - Diurnal and nocturnal differences in hypothalamic-pituitary-adrenal axis function in Galapagos marine iguanas. AB - Temporal modulation of the stress response is a ubiquitous characteristic of animals. Here, we investigate possible mechanisms underlying daily changes in corticosterone release in an ectotherm model system. Earlier work indicated that free-living Galapagos marine iguanas (Amblyrhynchus cristatus) have lower corticosterone concentrations during the night than during the day. This could result from: (i) a lower circadian secretion of adrenocorticotropic hormone (ACTH) as seen in mammals; (ii) from an increase in corticosterone negative feedback; or (iii) reflect lower metabolic activity during the night when core body temperature falls (from 35 degrees C during the day to as low as 21 degrees C during the night). To begin to distinguish between these three possibilities, exogenous ACTH was used to compare diel differences in adrenocortical tissue responsiveness, and dexamethasone was used to compare diel differences in the efficacy of corticosterone negative feedback. Low levels of exogenous ACTH (30 IU/kg body weight) potently stimulated both daytime and nighttime corticosterone release. Dexamethasone (1 mg/kg) inhibited only daytime, but not nighttime endogenous corticosterone release. Because the response to ACTH was similar between day and night we suggest that a simple lowering of core body temperature cannot explain the nighttime reduction in corticosterone release. However, the failure of negative feedback at night suggests that the response is not equivalent to the controlled downregulation seen in mammals. PMID- 16263125 TI - Testosterone modulates performance on a spatial working memory task in male rats. AB - Gonadal hormones have been shown to modulate memory retention in female rats. The current experiments examine the role of testicular hormones in modulating the performance of male rats on two spatial water maze tasks. In the first study, castrated and intact rats were trained on the visible platform and hidden platform versions of the Morris water maze task. Castration did not affect performance on either version of this reference memory task with castrated and intact rats demonstrating similar performance both during acquisition and on post training probe trials. In the second experiment, castrated and intact rats were tested on a delayed-matching-to-place version of the water maze. Rats received a series of trial pairs in the maze with a hidden platform located in the same pool location on the exposure and retention trials of each pair; between pairs of trials, however, the platform was repositioned to a novel pool location. The interval between trials was either 10- or 60-min and memory retention, taken as the difference between the pathlengths on the exposure and retention trials, declined as the interval increased. Relative to intact males, castrated males demonstrated impaired working memory retention at 60-min but not at 10-min retention intervals. This interval-dependent impairment in working memory retention was reversed by physiologic levels of testosterone replacement. These findings indicate that castration does not significantly affect acquisition or probe trial performance on a classic reference memory task but does impair spatial working memory retention, an effect that is reversed by exogenous testosterone. PMID- 16263126 TI - Comparative genomic hybridization analysis of spontaneous abortion. AB - OBJECTIVES: To evaluate the feasibility and superiority of comparative genomic hybridization (CGH) in the genetic analysis of spontaneously aborted tissues. METHODS: 38 conceptuses from early failed pregnancies were studied, of which, 27 samples were fresh and 11 were old. Each sample was divided into two parts, one part for conventional cytogenetic analysis and the other for CGH analysis. RESULTS: All 38 spontaneously aborted tissues were analyzed successfully by the CGH approach, but only 31 samples received results from the cytogenetic karyotype analysis, while 7 other tissues failed to get data due to failure in tissue culturing. Among the specimen successfully analyzed by both approaches, 90% (28 out of 31) obtained identical results, and 14 aneuploidies were found. The only structural chromosome aberration in this series, 46, XY, del(3) (q22-24), was found using the CGH approach, which appeared as a normal male karyotype on the chromosomal metaphase spread. Also, two cases indicated triploidies under cytogenetic analysis but appeared to be normal on the CGH profile. In addition, among the seven samples of tissue culture failure, CGH identified three to be aneuploidies. CONCLUSION: The CGH analysis accurately identifies chromosomal unbalanced abnormalities related to spontaneous abortions with low failure rate. PMID- 16263127 TI - Comparative in vitro study on the long-term performance of cemented hip stems: validation of a protocol to discriminate between "good" and "bad" designs. PMID- 16263128 TI - Changes in joint stability with muscle contraction measured from transmission of mechanical vibration. AB - A non-invasive in vivo technique was developed to evaluate changes in wrist joint stability properties induced by increased co-activation of the forearm muscles in a gripping task. Mechanical vibration at 45, 50 and 55 Hz was applied to the radial head in ten healthy volunteers. Vibrations of the styloid process of the radius and the distal end of the metacarpal bone of the index finger were measured with triaxial accelerometers. Joint stability properties were quantified by the transfer function gain between accelerations on either side of the wrist joint. Gain was calculated with the muscles at rest and at five force levels ranging from 5% to 25% of maximum grip force (%MF). During contraction the gain was significantly greater than in control trial (0%MF) for all contractions levels at 45 and 50 Hz and a trend for 15%MF and higher at 55 Hz. Group means of contraction force and gain were significantly correlated at 45 (R(2)=0.98) and 50 Hz (R(2)=0.72), but not at 55 Hz (R(2)=0.10). In conclusion, vibration transmission gain may provide a method to evaluate changes in joint stability properties. PMID- 16263129 TI - Impact of plastic adhesion in vitro on analysis of Th1 and Th2 cytokines and immune cell distribution from mice with multiple low-dose streptozotocin-induced diabetes. AB - Cytokines produced by Th1 or Th2 cells have been postulated to be important in the development of type 1 diabetes in humans and animal models, such as murine multiple low-dose streptozotocin (MLDSTZ)-induced diabetes. The aim of this study was to investigate cytokine production with or without in vitro depletion of plastic adherent cells from spleens isolated after MLDSTZ treatment. Spleen cells were prepared on day 14 from MLDSTZ- and saline-treated mice and divided into two fractions. One cell fraction was depleted of adherent cells by plastic adherence and the other was not. Both cell fractions were analysed by FACS for the distribution of immune cells. In other experiments, the cells were cultured for 48 h with concanavalin A stimulation. Supernatant samples were analysed by ELISA for TNFalpha, IFNgamma and IL-10 production. Either before or after the 48-h culture cytokine mRNA expression was determined by RT-PCR. Plastic adhesion decreased the macrophage numbers by approximately 30% and CD4(+)CD25(+) cells by about 60%. This was accompanied by increased medium levels of TNFalpha, IFNgamma and IL-10, which suggest that either CD4(+)CD25(+) cells, macrophages, or both, down-regulate production of both Th1 and certain Th2 cytokines. Depletion of adherent cells also decreased IL-4 mRNA amounts. MLDSTZ treatment increased the production of Th1 cytokines mainly at the protein level, and IL-10 mainly at the mRNA level. This indicates a sustained increase in Th1 production after MLDSTZ treatment and an increase in IL-10 that might reflect an attempt to counteract the MLDSTZ-induced immune damage. Plastic adhesion during cell preparation may affect the relative distribution of certain immune cells. PMID- 16263130 TI - Optimization of the modification of carrier proteins with aminated haptens. AB - In this report we show that succinic groups are far more reactive to amino compounds than the carboxylic groups derived from Asp and Glu on the protein when using coupling via 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDCI) (even by an 8 fold factor). Accordingly, a new carrier-protein was designed where both natural amino and carboxylic moieties were transformed into succinic residues. To prepare this hypersuccinylated carrier, all exposed carboxylic acids were first transformed into amino groups by reaction with ethylendiamine after activation with EDCI. Secondly, all these residues together with the ones from Lys were succinylated to prepare a fully succinylated protein. This was even more relevant considering that the amount of Lysine was 2-4 fold lower than Asp and Glu in most of the proteins. These "hyper-succinylated" proteins (KLH or BSA) offer significant improvements in protein reactivity compared to the native proteins (by a factor of 8-10). The optimization of the reaction, in which the presence of dioxane was found to be influential, permitted further improvements in the modification of the protein. Finally, this new strategy was successfully used to develop antibodies against the commercial anti-tumor molecule, ET-637-NH2. Using native KLH no response was found, whereas 1/64,000 serum dilutions gave very high values in ELISA procedures when immunization was performed using the hyper succinylated KLH. PMID- 16263132 TI - Fisherian and Wrightian perspectives in evolutionary genetics and model-mediated imposition of theoretical assumptions. AB - I investigate how theoretical assumptions, pertinent to different perspectives and operative during the modeling process, are central in determining how nature is actually taken to be. I explore two different models by Michael Turelli and Steve Frank of the evolution of parasite-mediated cytoplasmic incompatility, guided, respectively, by Fisherian and Wrightian perspectives. Since the two models can be shown to be commensurable both with respect to mathematics and data, I argue that the differences between them in the (1) mathematical presentation of the models, (2) explanations, and (3) objectified ontologies stem neither from differences in mathematical method nor the employed data, but from differences in the theoretical assumptions, especially regarding ontology, already present in the respective perspectives. I use my "set up, mathematically manipulate, explain, and objectify" (SMEO) account of the modeling process to track the model-mediated imposition of theoretical assumptions. I conclude with a discussion of the general implications of my analysis of these models for the controversy between Fisherian and Wrightian perspectives. PMID- 16263131 TI - Solid-substrate room-temperature phosphorescence immunoassay based on an antibody labeled with nanoparticles containing dibromofluorescein luminescent molecules and analytical application. AB - Luminescent 20-nm silicon dioxide nanoparticles containing dibromofluorescein (D; particles denoted D-SiO(2)) were synthesized by the sol-gel method. In the presence of Pb(Ac)(2) as a heavy atom perturber, the particle can emit the intense and stable room temperature phosphorescence(RTP) signal on a polyamide membrane at the wavelength of lambdaex(max)/lambdaem(max) = 460/623 nm for D. Our research indicates that the specific immune reaction between goat-anti-human IgG antibody labeled with D-SiO(2) and human IgG can be carried out quantitatively on a polyamide membrane, and the phosphorescence intensity was evidently enhanced after the immunoreaction. Thus a new solid substrate-room temperature phosphorescence immunoassay (SS-RTP-IA) for determination of human IgG was established. The linear range of this method is 0.0624-20.0 pg spot(-1) of human IgG (corresponding to a concentration range of 0.156-50.0 ng ml(-1), sample volume: 0.40 microl spot(-1)). The regression equation of the working curve is DeltaIp = 94.39 + 17.00 m IgG (pg spot(-1)) (460/623 nm, r = 0.9998). Detection limit calculated as 3 Sb/k is 0.015 pg spot(-1). After elevenfold replicate measurement, RSD are 3.2% and 2.4% for samples containing 0.156 and 50.0 ng l(-1) IgG, respectively. This method is sensitive, accurate, and of high precision. And it has been applied to the determination of IgG in human serum with satisfactory results. Meanwhile, the mechanism of SS-RTP-IA based on an antibody labeled with nanoparticles containing dibromofluorescein luminescent molecules was discussed. PMID- 16263133 TI - Dynamic regulation of single- and mixed-species malaria infection: insights to specific and non-specific mechanisms of control. AB - Our increasing appreciation of the high prevalence of mixed-species Plasmodium infection in malaria-endemic regions has resulted in controversy regarding the likely mechanism(s) of regulation for mixed parasite burden within an individual human host. In the present study, we examined dynamic models of Plasmodium spp. regulation by fever and by non-specific (NS) and species-specific (SS) immunity (including the influence of their variable time-delays, duration, and efficacy) in order to assess the likely role of these factors in regulating detectable parasitemia and clinical disease. Our models suggest that in order to observe the irregular waves of fever and parasitemia that are often found in multiply infected subjects, there must be a differential SS immune effect (beyond the regulatory effects of the species-transcendent density-dependent factors previously posited to control mixed-species parasitemia), and time-dependent variation in immunity to the dominant species. By implementation of individual SS immune controls of non-permanent duration, the resulting multi-dimensional model can be viewed as multiple single-species oscillators coupled via a NS species transcendent controller. This extended model exhibits the essential patterns of long-term mixed infections. Although this 'circuit-immunity' model gives only a qualitative estimate of the complex web of participating agents and reaction pathways, it provides a starting point for future studies of the specific and NS within-host mechanisms that regulate mixed-species malaria infection. PMID- 16263134 TI - Geographic profiling and animal foraging. AB - Geographic profiling was originally developed as a statistical tool for use in criminal cases, particularly those involving serial killers and rapists. It is designed to help police forces prioritize lists of suspects by using the location of crime scenes to identify the areas in which the criminal is most likely to live. Two important concepts are the buffer zone (criminals are less likely to commit crimes in the immediate vicinity of their home) and distance decay (criminals commit fewer crimes as the distance from their home increases). In this study, we show how the techniques of geographic profiling may be applied to animal data, using as an example foraging patterns in two sympatric colonies of pipistrelle bats, Pipistrellus pipistrellus and P. pygmaeus, in the northeast of Scotland. We show that if model variables are fitted to known roost locations, these variables may be used as numerical descriptors of foraging patterns. We go on to show that these variables can be used to differentiate patterns of foraging in these two species. PMID- 16263135 TI - Near-periodic substitution and the genetic variance induced by environmental change. AB - We investigate a model that describes the evolution of a diploid sexual population in a changing environment. Individuals have discrete generations and are subject to selection on the phenotypic value of a quantitative trait, which is controlled by a finite number of bialleic loci. Environmental change is taken to lead to a uniformly changing optimal phenotypic value. The population continually adapts to the changing environment, by allelic substitution, at the loci controlling the trait. We investigate the detailed interrelation between the process of allelic substitution and the adaptation and variation of the population, via infinite population calculations and finite population simulations. We find a simple relation between the substitution rate and the rate of change of the optimal phenotypic value. PMID- 16263136 TI - Neuronal expression of nuclear transcription factor MafG in the rat medulla oblongata after baroreceptor stimulation. AB - The medulla oblongata is the site of central baroreceptive neurons in mammals. These neurons express specific basic-leucine zipper transcription factors (bZIP) after baroreceptor stimulation. Previously we showed that activation of baroreceptors induced expression of nuclear transcription factors c-Fos and FosB in central baroreceptive neurons. Here we studied the effects of baroreceptor stimulation on induction of MafG, a member of small Maf protein family that functions as dimeric partners for various bZIP transcription factors by forming transcription-regulating complexes, in the rat medulla oblongata. To determine whether gene expression of MafG is induced by stimulation of arterial baroreceptors, we examined the expression of its mRNA by semi-quantitative reverse transcription-PCR method and its gene product by immunohistochemistry. We found that the number of MafG transcripts increased significantly in the medulla oblongata after baroreceptor stimulation. MafG-immunoreactive neurons were distributed in the nucleus tractus solitarii, the dorsal motor nucleus of the vagus nerve, the ambiguous nucleus and the ventrolateral medulla. The numbers of MafG-immunoreactive neurons in these nuclei were significantly greater in test rats than in saline-injected control rats. We also found approximately 20% of MafG-immunoreactive neurons coexpress FosB after baroreceptor stimulation. Our results suggest that MafG cooperates with FosB to play critical roles as an immediate early gene in the signal transduction of cardiovascular regulation mediated by baroreceptive signals in the medulla oblongata. PMID- 16263137 TI - Antioxidant activity of propionyl-L-carnitine in liver and heart of spontaneously hypertensive rats. AB - Oxidative stress plays an important role in arterial hypertension and propionyl-L carnitine (PLC) has been found to protect cells from toxic reactive oxygen species. In this work, we have evaluated the antioxidant capacity of chronic PLC treatment in spontaneously hypertensive rats (SHR) by measuring the activity of antioxidant enzymes and the lipid peroxidation in liver and cardiac tissues. The activity of glutathione peroxidase was decreased in liver and cardiac tissues of SHR when compared with their normotensive controls, Wistar- Kyoto (WKY) rats, this alteration being prevented by PLC treatment. Glutathione reductase activity was increased in hypertensive rats and no effect was observed after the treatment. No significant changes in superoxide dismutase activity were observed among all experimental groups. Liver of hypertensive rats showed higher catalase activity than that of normotensive rats, and PLC enhanced this activity in both rat strains. Thiobarbituric acid reactive substances, determined as a measure of lipid peroxidation, were increased in SHR compared with WKY rats, and PLC treatment decreased these values not only in hypertensive rats but also in normotensive ones. The content of carnitine in serum, liver and heart was higher in PLC-treated rats, but PLC did not prevent the hypertension development in young SHR. In addition, triglyceride levels, which were lower in SHR than WKY rats, were reduced by chronic PLC treatment in both rat strains. These results demonstrate: i) the hypotriglyceridemic effect of PLC and ii) the antioxidant capacity of PLC in SHR and its beneficial use protecting tissues from hypertension-accompanying oxidative damage. PMID- 16263138 TI - Effects of haloperidol and melatonin on the in situ activity of nigrostriatal tyrosine hydroxylase in male Syrian hamsters. AB - Haloperidol, an antipsychotic drug, was tested for its effects on the in situ activity of nigrostriatal and hypothalamic tyrosine hydroxylase, in control male Syrian hamsters and in those receiving a high daily dose of melatonin. After receiving daily ip injections (1.25 mg/kg ip) of haloperidol for 21 days, the animals were sacrificed and brain tissue collected for analysis of dopamine and metabolites by HPLC with electrochemical detection. In situ activity of tyrosine hydroyxlase (TH) activity was determined by measuring the accumulation of L-Dopa after administration of the L amino acid decarboxylase inhibitor, mhydroxybenzylhydrazine. Tissue content of dopamine and its metabolites, DOPAC and HVA, was depressed in striatum of animals receiving haloperidol, and tyrosine hydroxylase (TH) activity was significantly decreased 20-24 h after the last injection (from 1823 +/- 63 to 1139 +/- 85 pg l-dopa/mg tissue). The decrease in TH activity in striatum was significantly inhibited by daily injections of a high dose of melatonin (2.5 mg/kg ip) (from 1139 +/- 85 to 1560 +/- 116 pg L-dopa/mg tissue). In the substantia nigra and in the hypothalamus, on the other hand, haloperidol significantly increased the activity of tyrosine hydroxylase. Melatonin administration did not significantly influence TH activity in the substantia nigra, but inhibited TH activity in the hypothalamus and in the pontine brainstem. One explanation for these data is that chronic haloperidol administration in Syrian hamsters increases TH activity in hypothalamus and substantia nigra, but decreases TH activity in striatum by a mechanism involving D2 presynaptic receptors and a melatonin sensitive kinase which regulates TH phosphorylation. PMID- 16263140 TI - Prostate tumor CXC-chemokine profile correlates with cell adhesion to endothelium and extracellular matrix. AB - Though chemokines of the CXC family are thought to play key roles in neoplastic transformation and tumor invasion, information about CXC chemokines in prostate cancer is sparse. To evaluate the involvement of CXC chemokines in prostate cancer, we analyzed the CXC coding mRNA of both chemokine ligands (CXCL) and chemokine receptors (CXCR), using the prostate carcinoma cell lines PC-3, DU-145 and LNCaP. CXCR proteins were further evaluated by Western blot, CXCR surface expression by flow cytometry and confocal microscopy. The expression pattern was correlated to adherence of the tumor cells to an endothelial cell monolayer or to extracellular matrix components. Based on growth and adhesion capacity, PC-3 and DU-145 were identified to be highly aggressive tumor cells (PC-3>DU-145), whereas LNCaP belonged to the low aggressive phenotype. CXCL1, CXCL3, CXCL5 and CXCL6 mRNA, chemokines with pro-angiogenic activity, were strongly expressed in DU-145 and PC-3, but not in LNCaP. CXCR3 and CXCR4 surface level differed in the following order: LNCaP>DU-145>PC-3. The differentiation factor, fatty acid valproic acid, induced intracellular CXCR accumulation. Therefore, prostate tumor malignancy might be accompanied by enhanced synthesis of angiogenesis stimulating CXC chemokines. Further, shifting CXCR3 and CXCR4 from the cell surface to the cytoplasm might activate pro-tumoral signalling events and indicate progression from a low to a highly aggressive phenotype. PMID- 16263141 TI - Petroleum hydrocarbons, polycyclic aromatic hydrocarbons, organochlorine pesticides and polychlorinated biphenyls in tissues of Indo-Pacific humpback dolphins from south China waters. PMID- 16263139 TI - Direct antimicrobial activities of PR-bombesin. AB - PR-bombesin is a bombesin-like peptide derived from the skin of the Chinese red belly toad, Bombina maxima. The 8-residue segment of N-terminal of RP-bombesin, comprising four prolines and three basic residues, is extensively different from other bombesin-like peptides. Since sequence of Pro-Arg-Pro generally plays an important role in the antimicrobial activity of proline-rich antimicrobial peptides, the componential feature of PR-bombesin indicates that it may have antimicrobial activity. In this paper, we presented the first evidence that bombesin-like peptides possess direct antimicrobial activities as some neuropeptides. It was determined by CD spectroscopy that PR-bombesin adopted a combination of random coil and beta-sheet structure, suggesting RP-bombesin is a new member of antimicrobial peptides having beta structure but without disulfide bonds. Current results also supported that PR-bombesin plays a direct defensive role besides its neuro-endocrological functions. PMID- 16263142 TI - A hierarchical approach measures the aerial extent and concentration levels of PAH-contaminated shoreline sediments at historic industrial sites in Prince William Sound, Alaska. AB - A field study was conducted in 2003 to estimate the areal distribution and concentrations of polycyclic aromatic hydrocarbons (PAH) in intertidal sediments at sites of past human and industrial activity (HA sites) in Prince William Sound (PWS), Alaska, the site of the 1989 Exxon Valdez oil spill. More than 50 HA sites, primarily in western PWS, were identified through analysis of historic records and prior field studies, and nine sites were selected for detailed surveys. The areal assessment process consisted of seven steps: (1) identify site from historic records and field surveys; (2) locate visual evidence of surface oil/tar at a site; (3) prepare a site map and lay out a sampling grid over the entire site with 10-m grid spacing; (4) excavate pits to 50 cm depth on the grid; (5) perform a field colorimetric test to estimate total PAH (TPAH) in sediments from the wall of each pit and record the results in the ranges <1 ppm; 1-10 ppm; >10 ppm TPAH; (6) expand grid size if necessary if elevated PAH levels are detected colorimetrically; (7) select 20 samples from each site for same-day shipboard PAH analysis by immunoassay (SDI RaPID PAH) and, based on these results, select sediment samples from each site for full PAH analysis in the laboratory to identify PAH sources. A total of 416 pits were dug at the nine sites. Nine acres of sediments with TPAH >2500 ppb dry wt. were mapped at the nine sites. TPAH concentrations obtained by immunochemical analysis of 181 samples from the nine sites ranged from 20 to 1,320,000 ppb (wet wt.). The contaminants are mixtures of petroleum products (2-3 ring PAH) and combustion products (4-6 ring PAH) unrelated to the 1989 Exxon Valdez oil spill. Mussels and clams collected at these sites have elevated levels of PAH that are compositionally similar to the PAH in the sediments. These findings indicate that at least a portion of the sediment PAH is bioavailable. The PAH sources at these historic industrial sites are chronic. They include relict fuel oil tanks and works located above and within the intertidal zone, with contamination at some locations extending into nearshore sub-tidal sediments. This study shows how a hierarchical approach can be used to quickly and successfully map, quantify, and subsequently, identify sources of PAH in shoreline sediments. PMID- 16263143 TI - The use of environmental integrative indicators to assess seabed disturbance in estuaries and coasts: Application to the Humber Estuary, UK. AB - The coastal zone is subject to many and varied changes resulting from human activities and natural processes. Existing or emerging agreements and legislation acknowledge the relevance of indicators for monitoring these. In the UK, following a series of recent workshops, potential indicators of nearshore disturbance have been identified and grouped into three broad indices: 'Coastline Morphological Change', 'Resource Use Change' and 'Environmental Quality and its Perception'. The present study developed these indicators further and tested their use by applying them to 18 sections in the Humber Estuary, Eastern England. The results obtained reflect the current knowledge of the state of the Humber environment and show the potential of integrative indicators but indicate that further studies are required to assess the relative importance of the indicators and their value in reflecting the ability of the ecosystems to sustain natural habitats and populations at a good conservation status. PMID- 16263145 TI - The non-regulation of food intake in humans: hope for reversing the epidemic of obesity. AB - Few doubt that human feeding behavior is part of larger biology regulatory system of energy stores, but the extent to which eating behavior is controlled by these biology systems and how much is due to responses to environmental stimuli is presently under debate. The results of a series of studies are presented which have attempted to determine the responsiveness of human feeding behavior to some of the "classic" biological variables that have conventionally been used to argue the biological basis of eating behavior. When humans are challenged with either overfeeding, underfeeding, or alterations of the caloric density of the diet, they fail to demonstrate precise caloric compensation. When challenged with changes in environmental stimuli, on the other hand, humans appear to be very sensitive to changes in portion size, the number of people with whom they eat, the amount that others eat and the variety of foods available. Other more chronic influences demonstrate that body weight appears to change when people move from one area of the world to another, when they enter the college environment, or when they either marry or break up. It is argued that because humans appear to be more responsive to the external environment than internal biological cues, it should be possible to curb or even reverse the epidemic of obesity by changing aspects of the external environment or human interactions with environmental variables rather than changing their internal environment through pharmacology. PMID- 16263144 TI - Memory inhibition and energy regulation. AB - At a simple behavioral level, food intake and body weight regulation depend on one's ability to balance the tendency to seek out and consume food with the ability to suppress or inhibit those responses. Accordingly, any factor that augments the tendency to engage in food seeking and eating or that interferes with the suppression of these behaviors could produce (a) caloric intake in excess of caloric need; (b) increases in body weight leading to obesity. This paper starts with the idea that excess body weight and obesity stem from a failure or degradation of mechanisms that normally function to inhibit eating behavior. Unlike previous approaches, we focus not on failures of traditional physiological (e.g., neural, hormonal) regulatory control mechanisms, but on disruptions of inhibitory learning and memory processes that may help to regulate energy intake. This view of energy dysregulation as a type of "learning disorder" leads us to the hippocampus, a brain structure that has long been regarded as an important substrate for learning and memory and which we think may be critically involved with a specific type of memory inhibition function that could contribute to the suppression of food intake. With this focus, the search for environmental origins of the current obesity epidemic in Western populations is directed toward factors that alter hippocampal functioning. We conclude by offering a preliminary account of how consumption of foods high in saturated fats might lead to impaired hippocampal function, reduced ability to inhibit caloric intake and, ultimately, to increased body weight. PMID- 16263146 TI - Health and road transport in Pakistan. AB - OBJECTIVE: The 1998 Global Burden of Disease Study estimated that road traffic injuries (RTIs) will become the third leading cause of lost disability-adjusted life years, with two-thirds of the deaths occurring in the least developed nations. Moreover, automobile-based transport systems are associated with air pollution (lead toxicity, asthma and greenhouse gas accumulation), noise disturbances, physical inactivity and obesity. STUDY DESIGN: This study (1) reviewed road transport literature in Pakistan and the impacts on health outcomes; (2) examined health policies to assess their focus on transport-related health problems; and (3) identified policy gaps for future research. METHODS: A methodological review of the literature on direct and indirect effects of road transportation in Pakistan. This review includes government documents, memos, statements and draft policies as well as relevant articles indexed in MEDLINE. RESULTS: A systematic review revealed no approved transport policy in Pakistan, despite three national health policy documents. The Health Chapter of the 9th Five Year Plan appreciates the grave threat of unchecked RTI, but fails to offer specific policy interventions. Despite ambitious plans by the Pakistan Environmental Protection Agency, actual projects and their implementation remain scarce, resulting in ever-increasing air pollution. The health impact of lead toxicity, noise pollution and RTIs remain high, while obesity is on the rise. CONCLUSION: The increasing health impact of road transport on 140 million people calls for immediate policy action. Government agencies must intervene effectively to establish monitoring and decentralised enforcement nationwide, while simultaneously supporting alternative modes of transportation. PMID- 16263147 TI - Induction of hepatic cytochrome P450 isozymes, benzo(a)pyrene metabolism and DNA binding following exposure to polycyclic aromatic hydrocarbon residues generated during repeated fish fried oil in rats. AB - In the present study the effect of repeated fish fried oil (RFFO) and its extract (RFFE) on hepatic cytochrome P450 (CYP) isozymes, benzo(a)pyrene (BP) metabolism and DNA adduct formation was undertaken. HPLC analysis of RFFO showed the presence of several polycyclic aromatic hydrocarbons. CYP in microsomes from control and RFFO-treated animals showed a peak at 450 nm; however, a shift of 2 nm in the SORET region along with significant induction was observed in microsomes prepared from 3-methylcholanthrene (MC)- and RFFE-treated animals. Activities of hepatic ethoxyresorufin-O-deethylase, methoxyresorufin-O deethylase, aryl hydrocarbon hydroxylase and erythromycin-N-demethylase were found to be significantly (P < 0.05) induced following exposure of RFFE, whereas none of these enzymes were altered in RFFO-treated group. Immunoblot analysis revealed that RFFE and MC were potent inducers of CYP1A1, 1A1/2 and 3A1 isozymes, where as RFFO showed no change in these protein levels. RT-PCR analysis showed induction of cDNA of CYP1A1 and CYP3A1 by RFFE treatment. Hepatic microsomes prepared from RFFE exposed animals enhanced BP metabolism with a concomitant increase in the relative proportion of BP 7,8-diol. Hepatic microsomes prepared from animals pretreated with RFFE and MC significantly enhanced the binding of [(3)H]-BP to calf thymus DNA. The overall results suggest that exposure to RFFE may induce hepatic CYP isozymes thereby producing enhanced reactive metabolites with a potential to bind with DNA that may result in cancer. PMID- 16263148 TI - Characterization of the acute pancreatitis induced by secretory phospholipases A2 in rats. AB - Acute pancreatitis (AP) is an inflammatory disease of the pancreas characterized by local inflammation and extrapancreatic effects such as lung injury. Secretory phospholipases A(2) (PLA(2)s) have been implicated in triggering AP, but their exact role to evoke AP is largely unknown. Therefore, we have tested the ability of sPLA(2)s to induce AP in rats, using venom sPLA(2)s with residual or high enzymatic activity (bothropstoxin-II and Naja mocambique mocambique venom PLA(2), respectively), as well as sPLA(2) devoid of catalytic activity (piratoxin-I). The injection of Naja m. mocambique venom PLA(2), bothropstoxin-II or piratoxin-I (300 microg/kg each) into the common bile duct increased significantly the pancreatic plasma extravasation and myeloperoxidase activity. The lung myeloperoxidase and serum amylase were also increased for all groups, although the Naja mocambique mocambique venom PLA(2) induced higher lung myeloperoxidase and serum amylase values, compared with piratoxin-I and/or bothropstoxin-II. Histopathology of pancreas and lungs in piratoxin-I-injected rats showed interstitial oedema in both tissues, and neutrophil infiltration with acinar cell necrosis in pancreas. In conclusion, sPLA(2)s induce AP in rats and the catalytic activity is not essential to induce the local effects in pancreas, although it appears to contribute partly to the remote lung injury. PMID- 16263149 TI - Strain-specific P3 of Soybean mosaic virus elicits Rsv1-mediated extreme resistance, but absence of P3 elicitor function alone is insufficient for virulence on Rsv1-genotype soybean. AB - When challenged by mechanical inoculation, the Rsv1 gene of soybean invokes extreme resistance (ER) against Soybean mosaic virus (SMV) strain N, but not SMV G7 and its experimentally evolved variant, SMV-G7d. SMV-G7 provokes a lethal systemic hypersensitive response (LSHR), whereas SMV-G7d induces systemic mosaic. Thus, for Rsv1-genotype soybean, SMV-G7 and SMV-G7d are both virulent virus strains. The elicitor function of SMV-G7 provoking Rsv1-mediated LSHR was recently mapped to P3, and the influence of amino acids 823, 953, and 1112 of the precursor polypeptide of SMV-G7d on evasion of Rsv1-mediated recognition provoking LSHR was demonstrated. We have now extended this study to SMV-N. Initially, amino acids corresponding to those of SMV-G7d at these positions were substituted, individually or in combinations. All the mutants remained replication competent on rsv1-genotype soybean; however, none lost the elicitor function provoking Rsv1-mediated ER. Subsequently, P3 of SMV-N was precisely replaced with P3 of SMV-G7 or SMV-G7d and vice versa. All the chimeras were replication competent on rsv1-genotype soybean, but surprisingly SMV-N/G7P3 and SMV-N/G7dP3 failed to gain virulence on Rsv1-genotype soybeans. However, SMV G7/NP3 and SMV-G7d/NP3 lost virulence, and this loss of virulence function was mapped to the N-terminus domain of SMV-N P3. The data indicate that SMV strain specific P3 provokes Rsv1-mediated ER; however, virulence on Rsv1-genotype soybean is not solely a consequence of the absence of the P3 elicitor functions provoking Rsv1-mediated ER and LSHR. PMID- 16263150 TI - High throughput sequence analysis reveals hitherto unreported recombination in the genus Norovirus. AB - Viruses of the Norovirus genus (Caliciviridae family) are a major cause of human gastroenteritis. In some viruses, recombination is an important evolutionary process and therefore we should try to discover the quantity and characteristics of such events in Noroviruses. In order to identify recombination events, multiple sequence alignments were assembled from publicly available strains, and were tested using RAT, a recently developed software tool. Strains identified by RAT as putative recombinants were tested further, using a phylogenetic approach, the LARD software, and a Monte Carlo method, to gain additional support for their status. The identification of two previously described recombinants, WUG1 and Snow Mountain, was made. Furthermore, three instances of hitherto unreported recombination implicating Norovirus strains MD 145-12, Gifu'96 and Saitama U4 were found, with good statistical support for the latter two of these cases. Lordsdale-like viruses were highlighted as major contributors to recombination events during Norovirus evolution. Finally, the relevance of recombinants to the worldwide transmission of Norovirus is discussed. PMID- 16263151 TI - The relationship between number of striae of Retzius and their periodicity in imbricational enamel formation. AB - Imbricational crown formation times (ICFTs) estimated from the number of perikymata on tooth surfaces are error-prone because the number of days between adjacent perikymata varies across individuals and species, and is only visible within tooth microstructure. We investigated striae of Retzius (SR) numbers (analogous to perikymata numbers), SR periodicities (days between SR or perikymata), and ICFTs for a mandibular canine sample (n=49) from medieval Denmark. We tested the relationship between SR number and periodicity to determine whether regression formulae could be produced that would allow periodicity (and ICFTs) to be determined from surface perikymata numbers. Periodicities (range=7-11 days, mode=8) and SR numbers (range=142-257, mean=190.3, s.d.=27.5) were normally distributed; ICFTs were non-normal (mean=1,594 days, s.d.=65.7). We tested periodicity as a quadratic, linear, and log-log transform linear function of SR number and found an inverse relationship (quadratic: R2=0.9504; linear: R2=0.9138; log-log transform: R2=0.9418; all p<0.001) that allowed estimation of periodicity from SR or perikymata numbers in this population and tooth type. If periodicity and SR number are inversely related in other hominin taxa, studies that have estimated ICFT by multiplying perikymata number by a human modal periodicity value or made inferences about development based only on perikymata numbers may have introduced substantial error into their ICFT estimates and life history inferences. The inverse relationship is similar to that predicted by a model of SR formation in which the ICFT for a given tooth type and population is held constant and all combinations of periodicity and SR number result in the same ICFT. However, we found that lower periodicities had longer ICFTs and higher periodicities had shorter ICFTs than the model predicted, suggesting that the model may not reflect the real process, or that there are other factors (e.g., sample size, misclassification, sexual dimorphism) also affecting the relationship between periodicity and SR number. PMID- 16263152 TI - A diagnosis of crocodile feeding traces on larger mammal bone, with fossil examples from the Plio-Pleistocene Olduvai Basin, Tanzania. AB - Neotaphonomic studies have determined the patterns of bone damage created by larger mammalian carnivores when consuming mammalian carcasses. Typically, mammalian carnivores gnaw and break bones to various degrees in order to access marrow, grease, and brain tissue. In contrast, crocodiles attempt to swallow whole parts of mammal carcasses, inflicting in the process tooth marks and other feeding traces on some of the bones they are unable to ingest. Although crocodiles are major predators of larger mammals along the margins of protected tropical rivers and lakes, their feeding traces on bone have received little systematic attention in neotaphonomic research. We present diagnostic characteristics of Crocodylus niloticus damage to uningested mammal bones resulting from a series of controlled observations of captive crocodile feeding. The resulting bone assemblages are composed of primarily complete elements from articulating units, some of which bear an extremely high density of shallow to deep, transversely to obliquely oriented tooth scores over often large areas of the bone, along with shallow to deep pits and punctures. Some of the tooth marks (bisected pits and punctures, hook scores) have a distinctive morphology we have not observed to be produced by mammalian carnivores. The assemblages are also characterized by the retention of both low- and high-density bone portions, an absence of gross gnawing, and minimal fragmentation. Together, the damage characteristics associated with feeding by crocodiles are highly distinctive from those produced by mammalian carnivores. Modern surface bone assemblages along the Grumeti River in Tanzania's Serengeti National Park contain a mixture of specimens bearing damage characteristic of crocodiles and mammalian carnivores. Comparison of Plio-Pleistocene fossil bones from Olduvai Gorge, Tanzania, to bones damaged by captive and free-ranging Nile crocodiles reveals direct evidence of fossil crocodilian feeding from larger mammal bones associated with Oldowan stone artifacts. PMID- 16263153 TI - The burning question: does burning before flooding lower methyl mercury production and bioaccumulation? AB - Production of methyl mercury (MeHg) is elevated in new hydroelectric reservoirs because organic carbon stimulates methylation of inorganic mercury (Hg) stored in the terrestrial system. This can cause adverse health in fish and in organisms that eat fish. We expected that burning vegetation before flooding would decrease the amount of Hg and organic carbon and thereby lower MeHg production. We conducted a replicated field experiment to investigate the effects of burning vegetation and soil before flooding on MeHg production and bioaccumulation. Vegetation and soil were added to mesocosms in the following combinations: unburned vegetation and unburned soil (Fresh treatments), burned vegetation and unburned soil (Partial Burn treatments), and burned vegetation and burned soil (Complete Burn treatments). Controls had no added vegetation or soil. During combustion with propane torches, a large percentage of the total Hg (THg) and MeHg was lost from vegetation and soil. THg and MeHg concentrations were highest in the surface water of Fresh treatments, lower in Partial Burn treatments and lowest in Complete Burn treatments and controls. Differences in concentrations of MeHg in biota were consistent among treatments, but did not follow aqueous concentrations. On the final sample date, MeHg concentrations in biota of Controls and Partial Burn treatments were greater than in Complete Burn and Fresh treatments. The lack of relationship between MeHg in biota and MeHg in water may have been due to modification of the bioavailability of MeHg by dissolved organic matter as the ratios of MeHg in biota to water were inversely correlated with concentrations of dissolved organic carbon. Although burning before flooding decreased MeHg concentrations in the water, it did not lower MeHg accumulation in the lower food web. PMID- 16263154 TI - The dilution effect of equal volume mixing studies compromises confirmation of inhibition by lupus anticoagulants even when mixture specific reference ranges are applied. AB - INTRODUCTION: One of the recommended criteria for the laboratory diagnosis of lupus anticoagulants (LA) is demonstration of inhibitory activity. This is confirmed by performing mixing tests with normal plasma, usually in a 1:1 ratio, and demonstrating persistence of an abnormal clotting time in the screening test with significant confirmatory test reduction. However, the mixing with normal plasma can dilute the antibodies to undetectable levels and generate apparent negative results. No guidelines or consensus exist in how to interpret mixing study results. PATIENTS AND METHODS: The present study assessed the 1:1 mixing study results from 600 patients with a thrombotic history positive for LA demonstrated in neat plasma by individual assays, or combinations, of dilute Russell's viper venom time, dilute activated partial thromboplastin time, activated seven lupus anticoagulant assay and Taipan snake venom time, plus confirmatory tests. Mixing tests were assessed initially using locally derived neat plasma reference ranges and subsequently with mixture specific ranges. RESULTS: The mixture specific ranges had lower upper limits. Of the total LA positive results, 32.5% were positive in the mixing studies when neat plasma reference ranges were applied, and a further 11.2% demonstrated LA activity when using the mixture specific ranges. The remaining 56.3% had been diluted such that they did not elevate the screening test above the upper limit of normal or generated minimal prolongation with an insignificant difference between the screen and confirmatory test result sufficient to confirm LA activity. CONCLUSIONS: The significant impact of the dilution effect in 1:1 mixing studies emphasises the limitations of mixing studies as a vehicle for confirmation of inhibition by LA antibodies. PMID- 16263155 TI - F-18 fluorodeoxyglucose uptake in primary cervical cancer as an indicator of prognosis after radiation therapy. AB - PURPOSE: We evaluated the prognostic significance of tumor metabolic activity on pretreatment positron emission tomography using the glucose analog F-18 fluorodeoxyglucose (FDG-PET) in patients with carcinoma of the cervix undergoing radiotherapy with or without concurrent chemotherapy. METHODS: We studied 96 consecutive patients who underwent FDG-PET prior to initiation of radiotherapy with or without concurrent chemotherapy for carcinoma of the cervix. Pretreatment FDG uptake of the primary tumor was assessed with the semiquantitative standardized uptake value (SUV) and correlated with disease-free and overall survival. Survival was estimated by the Kaplan-Meier method. Univariate and multivariate analyses were performed with the logistic likelihood ratio test and the Cox proportional hazards model, respectively. RESULTS.: Five-year disease free survivals in patients with maximal SUV <10.2 and > or =10.2 were 71% and 52% (P = 0.0289) respectively, while overall survivals were 72% and 69% (P = 0.4), respectively. On multivariate analysis, lymph node metastasis on FDG-PET was found to be predictive of disease-free survival (P < 0.0001). Both the SUV for FDG and FIGO Stage I disease were found to be marginally predictive of disease free survival (P = 0.055 and P = 0.058, respectively). CONCLUSIONS: FDG uptake within primary cervical cancer, as measured by SUV, is predictive of disease-free survival in patients undergoing radiotherapy for cervical cancer. High FDG uptake may be useful in identifying patients who may require more aggressive initial therapy. PMID- 16263156 TI - M344 is a novel synthesized histone deacetylase inhibitor that induces growth inhibition, cell cycle arrest, and apoptosis in human endometrial cancer and ovarian cancer cells. AB - OBJECTIVE: Histone deacetylase inhibitors (HDACIs) can inhibit cell proliferation, induce cell cycle arrest, and stimulate apoptosis of cancer cells. METHODS: We investigated the effects of a novel synthesized HDACI, M344, on Ishikawa endometrial cancer cell line, SK-OV-3 ovarian cancer cell line, and normal human endometrial epithelial cells. Endometrial and ovarian cancer cells were treated with various concentrations of M344, and its effect on cell growth, cell cycle, apoptosis, and related measurements was investigated. RESULTS: 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays showed that all endometrial and ovarian cancer cell lines were sensitive to the growth inhibitory effect of M344, although normal endometrial epithelial cells were viable after the treatment with the same doses of M344 that induced growth inhibition of endometrial and ovarian cancer cells. Cell cycle analysis indicated that their exposure to M344 decreased the proportion of cells in the S-phase and increased the proportion in the G0/G1 phases of the cell cycle. Induction of apoptosis was confirmed by annexin V staining of externalized phosphatidylserine and loss of the transmembrane potential of mitochondria. This induction occurred in concert with altered expression of genes related to cell growth, malignant phenotype, and apoptosis. Furthermore, M344 treatment of these cell lines increased acetylation of H3 and H4 histone tails. CONCLUSIONS: These results raise the possibility that M344 may prove particularly effective in the treatment of endometrial cancers and ovarian cancers. PMID- 16263157 TI - Expression of Wilms tumor gene (WT1) in epithelial ovarian cancer. AB - OBJECTIVES: The identification of proteins that are selectively expressed in cancer and with potential to elicit an immune response is the first step towards antigen-specific immunotherapy. The Wilms tumor gene product (WT1) is inherently immunogenic and is now thought to be oncogenic. The aim of this study was to determine the expression of WT1 in epithelial ovarian cancer (EOC) and correlate with clinico-pathologic characteristics. METHODS: WT1 expression was examined using immunohistochemistry applied on a tissue microarray of normal tissues and a panel of 100 EOC tissues. The distribution of WT1 expression and clinico pathologic variables were analyzed. Survival probabilities were estimated by Kaplan-Meier method, and statistical significance was determined by the log-rank test. RESULTS: WT1 expression was observed in 78/100 of specimens. The predominant expression pattern was homogenous, occurring in 66/100 (66%) of WT1 positive specimens, while 12/100 (12%) demonstrated heterogeneous staining. In normal tissues, WT1 expression was noted in kidneys, splenic capsule, Sertoli cells of the testis, and granulosa cells of the ovary. The median follow-up of the patient population was 30 months. Patients with WT1-positive tumors tended to have a higher grade (P = 0.006) and stage (P = 0.002) of tumor. However, there were no significant differences in the distribution of patients with WT1-positive tumors in relation to disease-free and overall survival. CONCLUSIONS: Our data demonstrate that WT1 is expressed at high frequency in patients with EOC. Since WT1 demonstrates tissue-restricted expression and is inherently immunogenic, it could represent an attractive target for antigen-specific immunotherapy in EOC. PMID- 16263158 TI - EGF-stimulated migration in ovarian cancer cells is associated with decreased internalization, increased surface expression, and increased shedding of the urokinase plasminogen activator receptor. AB - OBJECTIVES: The EGFR is expressed in malignant ovarian tumor tissue, and tissue content of EGFR has been directly associated with poor prognosis in patients with ovarian cancer. The uPA system plays a role in pericellular proteolysis, cell migration, invasion, and is over-expressed in ovarian cancer. This study explored the effects of EGF on uPAR expression in the ovarian cancer cell line OVCAR-3. METHODS: We used OVCAR-3 cells and the following methods: cell migration assay, time-lapse video microscopy, real-time PCR, assays for cellular binding of 125I uPA and cellular degradation of 125I-uPA:PAI-1 complex, biosynthetic labeling using 35S-methionin, Western blot, Northern blot, and ELISAs for uPA, PAI-1, and uPAR. RESULTS: EGF up-regulates both protein and mRNA not only for uPAR, but also for the ligand uPA and its inhibitor PAI-1. Cell surface uPAR, in control as well as EGF-stimulated cells, is present only in the intact, not the cleaved, form. Ligand binding experiments showed an increase of endogenously occupied uPAR, whereas non-occupied receptor sites were not increased. In addition, EGF treatment resulted in decreased degradation of radiolabeled uPA:PAI-1 complex. This suggests decreased internalization of uPAR, since the complex is internalized together with uPAR. Like EGF, colchicine, which inhibits endocytosis, increased cell surface expression of uPAR. In addition, we found an immediate increase of uPAR after exposing the cells to EGF and this was accompanied by a transient increase of cell migration. The increase of cell surface uPAR in response to EGF is accompanied by increased release of the soluble form of uPAR (suPAR) to the medium as well as by increased cell migration. Both uPAR and suPAR increased in cells treated with the endocytosis inhibitor colchicine even though cell migration was inhibited, suggesting that the mechanism of uPAR shedding is not related to cell migration. CONCLUSION: Increased cell surface uPAR in response to EGF stimulation results from mobilization of uPAR from detergent-resistant domains, increased expression of uPAR mRNA, and decreased internalization and degradation of uPAR. Both the anti uPAR antibody R3, which inhibits binding of uPA, and the EGFR phosphorylation inhibitor Iressa inhibited cell migration in response to uPA as well as to EGF, suggesting that EGFR and uPAR are engaged in the same multiprotein assembly on the cell surface. PMID- 16263159 TI - Application of sexed semen technology to in vitro embryo production in cattle. AB - Use of sexed semen in conjunction with in vitro embryo production is a potentially efficient means of obtaining offspring of predetermined sex. For thousands of years, livestock owners have desired a methodology to predetermine the sex of offspring for their herds. The ability to sort individual sperm cells into viable X- and Y-chromosome-bearing fractions made producers' sex selection dreams reality in the 1990s and now semen can be sexed with greater than 90% accuracy with use of a flow cytometric cell sorter. Several concerns regarding the implementation of sexed semen technology include the apparent lower fertility of sorted sperm, the lower survival of sorted sperm after cryopreservation and the reduced number of sperm that could be separated in a specified time period. These issues are discussed in this review. There are also a number of issues that appear to influence the success rates of using sexed semen to produce bovine embryos in vitro. These issues include reductions in fertilization rates, lower cleavage rates, blastocyst rates and pregnancy rates, partial capacitation of the sperm, dilute sperm samples and sire variation. These subjects are also addressed in this paper. Finally, we will describe a recent field trial in which female Holstein embryos produced using the combined technologies of sex-selected semen and microfluidics were transferred either as single or bilateral twin embryos into beef cattle recipients, demonstrating these technologies' contributions to viable embryo production. The results indicate that large-scale transfer of in vitro produced, Holstein heifer embryos to beef recipients is a feasible production scheme. PMID- 16263160 TI - Modifying oocytes and embryos to improve their cryopreservation. AB - Numerous studies indicate that in vitro-produced bovine embryos do not survive cryopreservation as well as those produced in vivo. Furthermore, embryos cultured in vitro in the absence of blood serum are more cryotolerant than embryos cultured in media containing serum. Although in vivo-produced embryos are more cryotolerant, there appear to be breed differences. Most if not all of these observations are correlated with cytoplasmic lipid content of embryos; more and larger lipid droplets are associated with reduced cryotolerance. This review concerns strategies for modifying oocytes and embryos to increase cryosurvival. Reduction of cytoplasmic lipid content of embryos with phenazine ethosulfate (PES), a compound that oxidizes NADPH, even improved cryotolerance of bovine embryos cultured in the absence of serum. Whether cytoplasmic lipid content per se or associated changes in lipid composition of cell membranes are responsible for differences in cryotolerance is unknown. Increasing cholesterol content of membranes of sperm and oocytes via cholesterol-loaded cyclodextrin also appears to improve cryotolerance. While lipids have been emphasized and appear to be important, non-lipid aspects of cell composition also likely affect cryotolerance, and might be modified to improve cryotolerance. Additional research on mechanisms of variation in cryotolerance will be applicable to circumvent cryo-intolerance attributable to variation associated with the individual animal, breed, species, cell type, and factors such as nutrition and season of the year. PMID- 16263161 TI - Survival and meiotic competence of bovine oocytes originating from early antral ovarian follicles. AB - The aim of the present study was to examine the growth and survival in culture, and the subsequent meiotic competence, of bovine oocytes recovered from early antral ovarian follicles. Follicles isolated by microdissection of the ovarian slices were sorted into two size groups: (I) 0.2-0.5 mm diameter; and (II) 0.4 0.7 mm diameter. Group I follicles were cultured intact while in Group II, cumulus-oocyte complexes with pieces of parietal granulosa were dissected from the follicles and cultured. Follicles or cumulus-oocyte complexes with parietal granulose were embedded in collagen gel and cultured in TCM 199 supplemented with 3% BSA and 4 mM hypoxanthine for 14 days (Group I) or 7-10 days (Group II). After this, cumulus-oocyte complexes were recovered from the gel. Oocytes that had lost the majority of the cumulus were fixed immediately after recovery. Cumulus-oocyte complexes showing normal morphology were either fixed immediately or were subjected to IVM for an additional 24h, and then were fixed. At the end of the growth culture, 57.6% of the compact COCs in Group I follicles were preserved in the GV configuration, 16.7% had resumed meiosis, and 25.8% were degenerated or did not show detectable chromatin. After IVM, the proportion of oocytes resuming meiosis increased significantly (from 16.7% versus 42.7%; P < 0.05), and 9.1% of all oocytes had reached TI or MII. The isolated cumulus-oocyte complexes in Group II began creating follicle-like structures following 24 h of growth culture (7.1%). The proportion of these structures reached 50.8% on days 2-3, and then gradually decreased due to degeneration. On day 10 only 5.8% of cumulus-oocyte complexes were classified as intact. Of the cumulus intact oocytes recovered from the newly created follicle-like structures at 7-10 days, 54.7% were in the germinal vesicle stage, 31.0% underwent germinal vesicle breakdown, 14.3% were degenerated or the chromatin configuration was not detectable. After 24 h of IVM, 67.6% of oocytes had resumed meiosis, and 21.6% of all oocytes had reached TI and MII. These results show that isolated early follicles and cumulus-oocyte complexes from intact early antral follicles can grow in culture and can develop meiotic competence. PMID- 16263162 TI - The first dromedary (Camelus dromedarius) offspring obtained from in vitro matured, in vitro fertilized and in vitro cultured abattoir-derived oocytes. AB - Dromedary offspring have never been produced fully in vitro. We have previously demonstrated that embryos obtained by culture in semi-defined medium (mKSOMaa) have better in vitro development ability than those cultured with oviductal epithelial cells. The aim of the present experiment was to study the pregnancy rate after embryo transfer of in vitro-produced (IVP) dromedary embryos cultured in semi-defined modified medium (mKSOMaa). IVM/IVF procedures were conducted on six hundred and sixty four (664) cumulus oocytes complexes (COCs) aspirated from ovaries collected at a local slaughterhouse and cultured in vitro (38.5 degrees C; 5% CO2, and maximum humidity >95%). Maturation was completed by incubation in TCM-199 medium supplemented with 10% heat-treated Fetal Calf Serum (FCS), 10 ng/mL EGF, 1 microg/mL FSH, 1 microg/mL E2 and 500 microM cysteamine for 30 h. In vitro fertilization was performed using fresh semen (0.5 x 10(6) spermatozoa/mL in modified TALP-solution). Fertilized oocytes were cultured in mKSOMaa, under 38.5 degrees C, 5% CO2 and 90% N2 with maximum humidity (>95%). All IVC steps were done in seven replicates. The cleavage rate (two cells to blastocyst stage) was 64% (425/664) and the percentage of oocytes reaching the blastocyst stage was 23% (155/664). The hatching rate of blastocyst obtained after culture was 46% (71/155). Good quality hatched blastocysts (n = 66) were transferred individually to synchronized recipients. Pregnancy rates, determined by ultrasonography at 15, 60 and 90 days after embryo transfer (ET), were 38%, 32% and 27%, respectively. Out of 18 pregnant females 5 aborted between the fifth and seventh month of pregnancy and 13 females (20%) remained pregnant. After 385 days of pregnancy, the first healthy and normal male-dromedary offspring produced fully in vitro was born at a birth weight of 38 kg. More dromedary calves (n = 4) were born later on. The remaining pregnant females (n = 8) are due to calf within the next months. In conclusion, this is the first reported offspring in camelids obtained by transfer of embryos produced by IVM, IVF and IVC using abattoir-derived oocytes, fresh semen and culture in a semi-defined medium. PMID- 16263163 TI - The relationships of adolescent school-related deviant behaviour and victimization with psychological distress: testing a general model of the mediational role of parents and teachers across groups of gender and age. AB - Deviant behaviour and victimization at school have been consistently related to poor psychological adjustment in adolescents. This research explores the mediating role that parents and teachers have in adolescent psychological distress in 973 Spanish students aged 11-16 years old. Structural equation analyses results showed that adolescent deviant behaviour and victimization were positively related to psychological distress as seen by the total effects. However, while victimization was directly related to psychological distress, the association of deviant behaviour and psychological distress was mediated by adolescent-parent communication and adolescent-teacher relationships. Multigroup analyses showed that relationships among variables were not significantly different for groups of age and gender. PMID- 16263164 TI - Bioprosthetic tissue preservation by filling with a poly(acrylamide) hydrogel. AB - Glutaraldehyde (GA) fixation has been used for more than 40 years as the preferred treatment to suppress immunogenicity and increase durability of bioprosthetic tissues (BPT) used in heart valve prostheses. This fixative and its reaction products have, however, been implicated in the calcific degeneration and long-term failure of these devices. The current study investigates stabilization of BPT and the mitigation/prevention of calcification by filling aortic wall samples with a synthetic poly(acrylamide) (pAAm) hydrogel, with and without pre treatment with GA. Histological and gravimetric analysis showed full penetration of the acrylamide (AAm) into the fresh tissue, while only partial filling could be achieved with GA pre-fixed tissue. The observed decrease in amino-group content (0.157+/-0.012-0.123+/-0.021 micromol/mg, p<0.03) and corresponding increase in shrinkage temperature (67.2+/-0.8-78.1+/-1.8 degrees C, p<0.0001) when fresh tissue was filled, indicate the participation of tissue-amines in a process that leads to BPT crosslinking. These effects were much less pronounced when the tissue was pre-fixed with GA. Filling increased the tensile stiffness of fresh tissue (to levels half that of 0.2% GA fixed tissue), but decreased the stiffness of GA pre-fixed tissue. When compared to standard 0.2% GA fixed samples, fresh tissue filled with AAm showed 88% (p<0.0001) less calcification while exhibiting similar resistance toward degradation by protease. Filling did not result in significant decreases in calcification when the tissue was pre fixed with GA. PMID- 16263165 TI - Localization of vascular endothelial growth factor (VEGF) and its receptors VEGFR 1 and VEGFR-2 in bovine placentomes from implantation until term. AB - Interactions of vascular endothelial growth factor (VEGF) with its receptors VEGFR-1 and VEGFR-2 promoting angiogenesis have been described in placentation of human, mink and pig. The bovine placenta is multiplex, villous and synepitheliochorial due to migratory trophoblast giant cells (TGC). To determine the role of VEGF in bovine implantation and placentation, placentomes and interplacentomal areas from 33 cows from early implantation until near term were evaluated by immunohistochemistry. VEGF immunoreactivity was detected in fetal and maternal blood vessel tissues during implantation and throughout gestation, and in preimplantatory trophoblast cells and uterine epithelium. After implantation the immunoreaction was confined to TGC and uterine epithelium. An antibody against bovine VEGF revealed a strong reactivity in the stroma of maternal caruncular septa in early and mid-gestation, which distinctly decreased near term. In interplacentomal areas, VEGF was found in luminal and glandular epithelia as well as in trophoblast, with distinctly higher reactivity in giant cells. VEGFR-1 was observed in trophoblast and uterine epithelium around implantation. Later, in definite placentomes, VEGFR-1 was localized in TGC near the chorionic plate and in maternal endothelial cells in the center of the placentome. VEGFR-1 and VEGFR-2 were co-localized in uterine epithelium and trophoblast as well as in blood vessel tissue and uterine glands. The presence of VEGF, VEGFR-1 and VEGFR-2 at the feto-maternal interface and in vasculature indicates that in the bovine VEGF may have (1) classic functions in angiogenesis and vascular permeability, (2) growth factor properties, facilitating feto maternal exchange via paracrine action, (3) chemotactic activity on capillary endothelium, and (4) an autocrine influence on TGC migratory activity. PMID- 16263166 TI - Modeling treatment motivation in substance-abusing women with children. AB - OBJECTIVES: Children are often considered a primary motivator for women seeking substance abuse treatment. This study tested a model predicting treatment motivation in substance-abusing mothers. METHODS: This study was a secondary analysis of the Drug Absue Treatment Outcome Study (DATOS). It used structural equation modeling to describe factors influencing motivation for treatment. DATOS is a national study of substance abusers entering treatment. Treatment was provided by a sampling of community-based programs, free-standing hospitals, hospital units, county-funded programs, modified therapeutic communities, and criminal justice programs. The subsample of women with children under the age of 18 for whom custody of children could be determined was selected (n=1371). The variables comprising each factor were based on self-report, and standardized scales measuring level of drug involvement, psychological functioning, children, and a desire to stop using drugs were used. RESULTS: Drug involvement was positively related to poorer psychological functioning, child custody issues, and the desire to stop using drugs. Child custody issues had a negative influence, while poorer psychological functioning and a desire to stop using drugs positively influenced treatment motivation. CONCLUSIONS: The negative influence that children have on treatment motivation may reflect the practical or emotional difficulties of having to leave children behind or in some instances having children placed in foster care. Specifically, losing custody of children, particularly with little expectation they will be reunified, may serve as a detriment to motivation. Popular beliefs hold that children serve as a primary source of a mother's treatment motivation; however this study found the opposite was true. Children should not automatically be considered a primary source of motivation for participation in treatment. PMID- 16263167 TI - Overreporting and underreporting of child abuse: teachers' use of professional discretion. AB - OBJECTIVE: According to mandatory reporting laws for professionals, the relationship between initial recognition that a child may have been abused and the subsequent reporting of that suspected case of child abuse to the responsible agency would, at first glance, appear to be clear. However, this relationship has developed into one of the major social policy controversies of the recent past. Our major goal is to present research findings that address this social policy debate concerning the problems of underreporting and overreporting, focusing specifically on teachers. METHOD: A factorial survey design, that combines the advantages of the factorial experiment with those of surveys, was employed in a probability sample of teachers (N=480) who responded to vignettes in which case characteristics were systematically manipulated. Teachers responded with judgments about whether the vignette was child abuse and the likelihood that they would report this suspected case. Characteristics of the teachers and their work setting (school) were also measured. RESULTS: When comparing the teachers' recognition and reporting scores, we found that they gave the same score for 63% of the vignettes they judged, gave higher reporting than recognition scores (overreporting) for 4% of the vignettes, and gave higher recognition than reporting scores (underreporting) for 33% of the vignettes. Discrepancies between recognition and reporting (over and under reporting) were related to characteristics of the case, teacher, and school where the teacher was employed. CONCLUSIONS: Teachers in our Ohio sample evidence the use of professional discretion in making judgments about the recognition and reporting of child abuse and do not appear to make these judgments with equal certainty. Their use of discretion is more likely to result in underreporting than overreporting. PMID- 16263168 TI - Sexual violence and youth in South Africa: the need for community-based prevention interventions. AB - OBJECTIVES: South Africa is reported to have one of the highest rates of sexual violence in the world, with adolescent girls between the ages of 12-17 being particularly at risk. Given that adolescence is considered a critical developmental period for establishing normative sexual behavior, this study explored multiple levels of risk influences that render adolescent girls vulnerable to becoming victims of sexual violence and adolescent boys vulnerable to becoming perpetrators of such abuse in one South African community. METHOD: A case study approach using qualitative rapid focused ethnographic methods was used. This involved 10 focus group interviews and 10 individual interviews with a volunteer convenience sample of adolescent boys and girls between the ages of 14 and 16 years. RESULTS: Inductive thematic analysis revealed that there were indeed multiple levels of risk influences for adolescent girls and boys becoming either victims or perpetrators of sexual violence. Using the Theory of Triadic Influence as a framework, influences at the distal socio-cultural/environmental level included traditional notions of masculinity and normalization of inter personal violence as well as poverty and the commodification of sex leading to rape supportive attitudes. Influences at the proximal situation context/social normative level included high-risk social norms as well as a weak adult and community protective shield. Finally, influences at the intra-personal level included low self-esteem and self-efficacy as well as inter-personal affective anger. CONCLUSION: Given the multiple levels of risk influences that need to be addressed to protect youth from becoming either perpetrators or victims of sexual violence in the South African context, prevention programs should necessarily be comprehensive, developmentally timed, and community-based. PMID- 16263169 TI - Albumin and amino acids upregulate the expression of human beta-defensin 1. AB - Antimicrobial peptides are essential components of the innate immune system and are the first line of defense against invading pathogens. Human beta-defensin 1 (hBD-1) is the most important antimicrobial peptide in human epithelia, its expression is constitutive in most tissues, and it is not induced in instances of infection or inflammation. In addition to its antibacterial activity, hBD-1 has an immunomodulatory activity by recruiting immune cells. Our objective was to determine whether immune-enhancing ingredients, such as arginine, isoleucine, and polyunsaturated fatty acids (PUFA), which are added to immune-enhancing formulas, mediate their beneficial activity by bolstering the immune system through hBD-1 induction. Real-time PCR analysis of hBD-1 in human colon cells, HCT-116, revealed upregulation after treatment with arginine, isoleucine, and bovine serum albumin (BSA). BSA increased transcription as well as secretion of hBD-1. hBD-1 upregulation in response to BSA treatment was concomitant with c-myc over expression, suggesting that hBD-1 may be regulated via a non-inflammatory pathway involving c-myc. Assuming similar control in vivo, our results may imply that nutrients found in immune-enhancing formulas upregulate hBD-1 expression, which, in turn, recruits the adaptive immune system. This sequence of events bolsters of the immune system, which may lead to fewer infectious complications, less antibiotics usage, and a shorter hospital stay, as indeed has been shown in patients fed immune-enhancing formulas. PMID- 16263170 TI - The catalytic activity of REV1 is employed during immunoglobulin gene diversification in DT40. AB - REV1 plays a key role in vertebrate translesion synthesis. Although its deoxycytidyl transferase activity is dispensable for tolerance of DNA damage caused by a number of mutagens, its extreme C terminus, which interacts with other translesion polymerases and PCNA, is essential. By examining immunoglobulin diversification in the genetically tractable chicken cell line DT40 we show that the generation of non-templated point mutations from C/G to G/C does require the catalytic activity of REV1. This provides the first clear evidence that the catalytic activity of REV1 is utilised in vivo in higher eukaryotes and is involved in immunoglobulin diversification. Although rev1 DT40 cells incorporate few point mutations, a mutant lacking the C terminus of REV1 exhibits a similar level to that seen in wild-type cells. Thus, the polymerase selection or stabilisation role of REV1 does not appear to play a major role in the bypass of AID-dependent abasic sites. PMID- 16263171 TI - Porcine Toll-like receptor 1, 6, and 10 genes: complete sequencing of genomic region and expression analysis. AB - Toll-like receptors (TLRs) recognize various microbial components and play key roles in activating the innate immune system. Hence, their function is important in swine infectious diseases. We completely determined 173,804 bp of nucleotide sequence of a genomic region including porcine TLR6 and the newly identified porcine TLR homologues TLR1 and TLR10. The porcine genomic structure of these genes was highly conserved in comparison with the corresponding region in humans. Analysis of their expression in porcine tissues showed differences in expression patterns between porcine TLR10 and TLR1 or TLR6. Moreover, phylogenetic analysis of the cytoplasmic regions of TLR genes suggested that the signal transduction pathway of TLR10 was different from those of TLR1 and TLR6. We also developed six polymorphic microsatellite markers within this genomic region; these markers will be valuable for association studies between TLR genes and resistance or susceptibility to infectious diseases in swine. PMID- 16263172 TI - Lipopolysaccharide-induced production of interleukin-10 is promoted by the serine/threonine kinase Akt. AB - The bacterial endotoxin lipopolysaccharide (LPS), is a potent inducer of the inflammatory response. Previous studies demonstrated that LPS-induced toxicity is reversed upon FcgammaR clustering by IgG immune complexes (IC) through upregulation of the anti-inflammatory cytokine IL-10. The PI3K-Akt pathway is also reported to reverse LPS-induced inflammation. In this study, we have examined the role of Akt in LPS-induced IL-10 production. First, we compared Akt activation in macrophages stimulated with either LPS alone, or with a combination of LPS and ICs. Our experiments revealed that while Akt was activated under both conditions, the level of activation was significantly higher in cells stimulated with LPS and ICs, suggesting that Akt may be involved in IC-induced upregulation of IL-10 production. Using several independent models we have then tested the notion that enhanced Akt activation may lead to enhanced LPS-induced IL-10 production. Over-expression of constitutively active Myr-Akt in the mouse macrophage cell line Raw 264.7 led to significant increase in IL-10 production in response to LPS. In addition, down-regulation of Akt by siRNA resulted in a decrease in LPS-induced IL-10 production. Peritoneal macrophages from transgenic mice with macrophage-specific expression of Myr-Akt produced significantly higher levels of IL-10 when stimulated with LPS, compared to their wild-type counterparts. Consistent with this observation, serum levels of IL-10, post-LPS challenge, was higher in the Myr-Akt transgenic mice compared to the wild-type mice. Taken together, these data demonstrate that Akt plays a critical role in LPS-induced production of IL-10. PMID- 16263173 TI - Localization of the third heparin-binding site in the human complement regulator factor H1. AB - Complement factor H (fH) plays a pivotal role in regulating the alternative pathway, allowing complement activation to proceed on foreign surfaces, whilst protecting surrounding host cell surfaces from complement-mediated damage. Host cell recognition is mediated by polyanions such as sialic acid and glycosaminoglycans (GAGs), which promote a high affinity interaction between fH and C3b deposited on host cell surfaces. Factor H is composed of 20 short consensus repeats (SCRs); two heparin-binding sites have been identified within SCR 7 and SCR 20 and a third site is thought to exist within or near SCR 13. Using an extensive series of recombinant fH fragments and heparin affinity chromatography, we have localized the third heparin-binding domain to SCR 9. A recombinant fH fragment containing both SCR 7 and SCR 9 exhibited higher affinity for heparin than SCR 7 alone, suggesting that the individual heparin-binding sites interact simultaneously with heparin to create a higher avidity interaction. Recombinant fragments containing SCR 9 bound to endothelial cells, indicating that this domain is capable of interacting with polyanions within a physiologically relevant environment. In addition, the three heparin-binding sites exhibited differences in their specificity for certain GAGs, suggesting that the individual binding domains may possess separate GAG recognition functions. PMID- 16263176 TI - Can herbs provide a new generation of drugs for treating Alzheimer's disease? AB - The overall aim of this review is to discuss cellular mechanisms at work in the progression of AD and current therapeutic strategies for treating AD, with a focus on the potential efficacy of herbal treatments. Recent advances in molecular, cellular, and animal model studies have revealed that formation of the 4-kDa amyloid beta peptide is a key factor in the development and progression of AD. Several cellular changes have been identified that are related to amyloid beta plaques and neurofibrillary tangles found in the autopsied brains of AD patients and in AD animal models. Several therapeutic strategies have been developed to treat AD, including anti-inflammatory, anti-oxidant, and anti amyloid approaches. Recently, herbal treatments have been tested in animal and cellular models of AD and in clinical trials with AD subjects. In AD animal models and cell models, herbal extracts appear to have fewer adverse effects than beneficial effects on A beta and cognitive functions. These extracts have multi functional properties (pro-cholinergic, anti-oxidant, anti-amyloid, and anti inflammatory), and their use in the treatment of AD patients looks promising. The chemical compositions of herbs and their potential for alleviating or reducing symptoms of AD or for affecting the disease mechanism need to be further studied. PMID- 16263174 TI - A 12-week, randomized, double-masked, multicenter study of the fixed combination of latanoprost and timolol in the evening versus the individual components. AB - PURPOSE: To compare the efficacy and tolerability of fixed-combination latanoprost and timolol applied in the evening with the concomitant use of the individual components. DESIGN: Twelve-week, randomized, double-masked, multicenter study. PARTICIPANTS: Five hundred seventeen randomized patients with ocular hypertension; open-angle, pigmentary, or exfoliation glaucoma; and baseline (after washout) intraocular pressure (IOP) levels between 23 and 33 mmHg. METHODS: Patients received either the fixed combination of latanoprost and timolol once daily in the evening and a placebo in the morning and evening or the unfixed combination of latanoprost once daily in the evening and timolol in the morning and evening. Study visits were at weeks 2, 6, and 12. MAIN OUTCOME MEASURES: The primary efficacy end point was mean change from baseline to week 12 in diurnal IOP (mean IOPs of 8 am, 12 pm, and 4 pm). The fixed combination was considered noninferior to the unfixed combination if the upper limit of the 95% confidence interval (CI) of the difference was <1.5 mmHg (analysis of covariance). Adverse events were recorded at each visit. RESULTS: In all, 502 patients were included in intent-to-treat analyses (fixed combination, n = 255; unfixed combination, n = 247). For the fixed- and unfixed-combination groups, mean baseline diurnal IOP levels were 25.4 mmHg and 25.2 mmHg, respectively, and mean diurnal IOP reductions were 8.7 mmHg and 9.0 mmHg (between-treatment difference, 0.3 mmHg; 95% CI, -0.1 to 0.7 mmHg; P = 0.15). Both treatments were well tolerated. CONCLUSIONS: The fixed combination of latanoprost and timolol administered once daily in the evening is not inferior to the unfixed combination of latanoprost once daily in the evening and timolol twice daily. The fixed combination provides an effective and well-tolerated alternative to multiple instillations. PMID- 16263177 TI - A comparative trial of psychotherapy and pharmacotherapy for "pure" dysthymic patients. AB - BACKGROUND: Psychotherapy of "pure" dysthymic disorder remains understudied. This article reports outcomes of an acute randomized trial of 94 subjects treated for 16 weeks with either interpersonal psychotherapy (IPT), brief supportive psychotherapy (BSP), sertraline, or sertraline plus IPT. METHODS: Recruited by clinical referral and advertising, subjects met DSM-IV criteria for early onset dysthymic disorder, with no episode of major depression in the prior six months. They were randomly assigned to one of four 16-week treatments, with options for crossover or continuation treatment. Results were analyzed from the intention-to treat sample by ANCOVA, controlling for baseline depressive severity. RESULTS: Subjects improved in all conditions over time, with the cells including sertraline pharmacotherapy showing superiority over psychotherapy alone for response and remission. Response rates were 58% for sertraline alone, 57% for combined treatment, 35% for IPT, and 31% for BSP. LIMITATIONS: The study was underpowered and may have employed too "active" a control condition. Follow-up data were unobtainable. CONCLUSIONS: In this acute trial for "pure" dysthymic disorder, sertraline with or without IPT showed advantages relative to IPT and BSP. Methodological difficulties may have limited differential outcome findings. This study bolsters a small but growing literature on the treatment of dysthymic disorder, suggesting that pharmacotherapy may acutely benefit patients more than psychotherapy. PMID- 16263178 TI - Low HDL cholesterol is associated with suicide attempt among young healthy women: the Third National Health and Nutrition Examination Survey. AB - BACKGROUND: Serum cholesterol is reported to be associated with suicidality, but studies conducted among general healthy population are rare. We examined the association between serum cholesterol and suicidality in a national sample of the general population of US. METHODS: We used the data of 3237 adults aged 17 to 39 years, who completed a mental disorder diagnostic interview and had blood specimens collected after a 12-h fast, as a part of the Third National Health and Nutrition Examination Survey, 1988-1994. The serum concentrations of total cholesterol, high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) were dichotomized according to the recommended levels of the National Cholesterol Education Program. A polytomous logistic regression was employed to control for covariates. RESULTS: Independent of socio demographic variables, health risks and nutrition status, and a history of medical and psychiatric illness (including depression), a significant association between low HDL-C (< or = 40 mg/dl) and increased prevalence of suicide attempts was observed in women (OR=2.93, 95% CI=1.07-8.00). No significant evidence was found to support an association between cholesterol and suicide ideation in women. Serum cholesterol was unrelated with either suicide ideation or attempts in men. LIMITATION: The inherent limitation of cross-sectional design prevented the authors from investigating causality. CONCLUSIONS: Low HDL-C is significantly associated with suicide attempts in women. Further studies are necessary to explore the clinical application of serum cholesterol as an indicator for suicide attempts among high risk population. PMID- 16263180 TI - Molecular cloning and characterization of bovine P-selectin glycoprotein ligand 1. AB - Human P-selectin glycoprotein ligand-1 (PSGL-1) is a dimeric membrane mucin expressed on leukocytes that binds selectins. Here, we report that the open reading frame (ORF) of bovine PSGL-1 (bPSGL-1) cDNA is 1284 base pairs in length, predicting a protein of 427 amino acids including an 18-amino-acid signal peptide, an extracellular region with a mucin-like domain, and transmembrane and cytoplasmic domains. The amino acid sequence of bPSGL-1 demonstrated 52, 49 and 40% overall homology to equine, human and mouse, respectively. A single extracellular cysteine, at the transmembrane and extracellular domain junction, suggests a disulfide-bonding pattern. Alignment of bovine with equine, human and mouse PSGL-1 demonstrates high conservation of transmembrane and cytoplasmic domains, but diversity of the extracellular domain, especially in the anionic NH(2)-terminal of PSGL-1, the putative P-selectin binding domain. In the NH(2) terminal of bPSGL-1, there are three potential tyrosine sulfation sites and three potential threonine O-glycosylation sites, all of which are required for P selectin binding in human PSGL-1 (hPSGL-1). bPSGL-1 shares only 57% homology in amino acid sequence with the corresponding epitope region which binds the monoclonal antibody PL1 for hPSGL-1, and no cross-reactivity was found in bovine leukocytes. In summary, bPSGL-1 shares homology with hPSGl-1, but has differences in the putative extracellular P-selectin binding domain. PMID- 16263179 TI - Modulation of juvenile brook trout (Salvelinus fontinalis) cellular immune system after Aeromonas salmonicida challenge. AB - In fish, the first line of defence against infectious microorganisms is based on non-specific cellular immune mechanisms (innate immunity). In this study, we measured the non-specific immune parameters (natural cytotoxic cells (NCC) activity, lymphoproliferation, percentage of phagocytosis and phagocytic activity) in brook trout (Salvelinus fontinalis) infected by a virulent strain of Aeromonas salmonicida. Eight days post-infection, the mortality of infected fish reached 70%. A transient immunostimulation of the NCC activity was noticed 24h post-infection, but there was no significant difference at 48 h. Then, infection of brook trout with A. salmonicida induced a biphasic immune response. At 24h post-infection, lymphoproliferation was drastically depressed but returned to control level at 96 h. A slight increase in the percentage of phagocytosis and the phagocytic activity was noticed throughout the experiment. Conversely the cell mortality was significantly higher in infected fish compared to control. The modulation of immunological parameters might reveal important clues on how innate immunity might protect fish from bacterial infections. PMID- 16263181 TI - No association of leukemia inhibitory factor (LIF) DNA polymorphisms with multiple sclerosis. AB - Neuropoietins such as leukemia inhibitory factor (LIF) have been shown to ameliorate experimental autoimmune encephalomyelitis (EAE) and promote oligodendrocyte survival in vivo. We tested whether two previously described LIF polymorphisms are associated with MS by genotyping these single nucleotide polymorphisms (SNPs) in a group of MS patients (n=110), rheumatoid arthritis (RA) patients (n=120) and healthy controls (HC, n=109). Similar allele and genotype frequencies for both SNPs were found for all study groups. Furthermore, no associations with MS type or HLA-DR2 expression could be found. In summary, no association was found between the studied LIF DNA polymorphisms and the prevalence of MS indicating that these polymorphisms are not involved in determining disease susceptibility. PMID- 16263182 TI - Prolonged prenatal exposure to low-level ozone affects aggressive behaviour as well as NGF and BDNF levels in the central nervous system of CD-1 mice. AB - The long-term effects on isolation-induced aggressive behaviour and central NGF and BDNF levels of gestational exposures to ozone (O(3)) were evaluated in adult CD-1 mice. Females were exposed to O(3), at the dose of 0.0, 0.3 or 0.6 ppm from 30 days prior the formation of breeding pairs until gestational day 17. Litters were fostered at birth to untreated dams and, at adulthood, male offspring underwent five successive daily encounters (15 min each) with a standard opponent of the same strain, sex, weight and age. The encounters on day 1, 3 and 5 were videotaped and agonistic and non-agonistic behavioural items finely scored. O(3) exposed mice showed a significant increase in freezing and defensive postures, a decrease in nose-sniffing behaviour and reduced progressively the aggressive behavioural profile displayed on day 1. Reduced NGF levels in the hippocampus and increased BDNF in the striatum were also found upon O(3) exposure. PMID- 16263183 TI - Bromophenols, both present in marine organisms and in industrial flame retardants, disturb cellular Ca2+ signaling in neuroendocrine cells (PC12). AB - Bromophenols are present in polychaetes as well as in algae in marine environments including the North Sea. They are thought to cause the typical sea like taste and flavour. The ecological function of brominated phenols is not clear yet, but they may play a role in chemical defence and deterrence [Kicklighter, C.E., Kubaneck, J., Hay, M.E., 2004. Do brominated natural products defend marine worms from consumers? Some do, most don't. Limnol. Oceanogr. 49, 430-441]. Some brominated phenols are commercially used as industrial flame retardants as, e.g., 2,4,6-tribromophenol and are suspected to disrupt the humoral system by showing thyroid hormone-like activity [Legler, I., Brouwer, A., 2003. Are brominated flame retardants endocrine disruptors? Environ. Int. 29, 879 885]. In this study 2-bromophenol (2-BP), 4-bromophenol (4-BP), 2,4-dibromophenol (2,4-DBP), 2,6-dibromophenol (2,6-DBP) and 2,4,6-tribromophenol (2,4,6-TBP), all of which are present in marine organisms, were tested. Especially 2,4-DBP and 2,4,6-TBP showed a significant effect on the Ca2+ homeostasis in endocrine cells (PC 12). The reduction of depolarization induced Ca2+ elevations by 2,4-DBP and 2,4,6-TBP and the increase of intracellular Ca2+ by both substances, partly released from intracellular stores, may suggest a link to the disrupting effect of endocrine systems by brominated phenols. 2,4-DBP was the most potent substance we tested in respect to inhibition of voltage dependent Ca2+ currents as revealed in whole cell patch clamp experiments. Brominated phenols disturb cellular Ca2+ signaling with differential efficacy, depending on the number and position of bromine. PMID- 16263186 TI - Structure and expression of the zebrafish mest gene, an ortholog of mammalian imprinted gene PEG1/MEST. AB - PEG1/MEST is a paternally expressed gene in placental mammals. Here, we report identification of zebrafish (Danio rerio) gene mest, an ortholog of mammalian PEG1/MEST. Zebrafish mest encodes a polypeptide of 344 amino acids and shows a significant similarity to mammalian orthologs. Zebrafish mest is present as a single copy in the zebrafish genome and is closely linked to copg2 as in mammals. It is notable that 10 of 11 intron positions in mest are conserved among mammalian PEG1/MEST genes, indicating that the genomic organization and linkage between mest and copg2 loci was established in ancient vertebrates. Zebrafish mest is expressed in blastula, segmentation, and larval stages, exhibiting gradually increased expression as the development proceeds. Allelic expression analysis in hybrid larvae shows that both parental alleles are transcribed. We also observed one-codon alternative splicing involving an alternative usage of the two consecutive splice acceptors of intron 1, generating two protein isoforms with different lengths of a single amino acid. PMID- 16263185 TI - Apelin: a new plasma marker of cardiopulmonary disease. AB - OBJECTIVES: Dyspnea is a major symptom of both parenchymal lung disease and chronic heart failure. Underlying cardiac dysfunction can be assessed by measurement of cardiac-derived B-type natriuretic peptide or its precursor in plasma. However, no specific endocrine marker of the lung parenchyma has so far been identified. We therefore examined whether plasma concentrations of apelin, a novel inotropic hormone, is affected in patients with chronic parenchymal lung disease without cardiac dysfunction. METHODS AND RESULTS: Patients with severe chronic parenchymal lung disease and normal cardiac function (n=53), idiopathic pulmonary hypertension with increased right ventricular pressure (n=10), and patients with severe left ventricular systolic dysfunction (n=22) were enrolled. Plasma apelin-36 and proBNP concentrations were measured with radioimmunoassays. While proBNP plasma concentrations were unaffected in chronic parenchymal lung disease patients compared to normal subjects, the apelin-36 concentration was reduced 3.3-fold (median 35 pmol/l (0-162 pmol/l) vs. 117 pmol/l (55-232 pmol/l), P<0.001). Moreover, the apelin-36 concentration was decreased in chronic heart failure patients (2.1-fold, P<0.01) and in patients with idiopathic pulmonary hypertension (4.0-fold, P<0.001). In contrast, the proBNP concentration was highly increased in both chronic heart failure and idiopathic pulmonary hypertension patients. CONCLUSION: Plasma concentrations of apelin-36, a novel inotropic peptide, are decreased in patients with chronic parenchymal lung disease and preserved cardiac function. Combined measurement of apelin-36 and proBNP may be a new diagnostic approach in distinguishing pulmonary from cardiovascular causes of dyspnea. PMID- 16263184 TI - Differential detoxification and antioxidant response in diverse organs of Corydoras paleatus experimentally exposed to microcystin-RR. AB - We assessed changes in activities of both detoxification and antioxidant enzymes as well as lipid peroxidation levels in liver, gill, intestine and brain of Corydoras paleatus exposed to dissolved microcystin-RR (MC-RR). Fish were captured at an unpolluted area, transported to the laboratory, and acclimated previous to experiments. Exposures were carried out using MC-RR at 0.5, 2, 5 and 10 microg L(-1). After exposures for 24h, fish were sacrificed and dissected separating liver, gills, intestine and brain of each fish. Organs were used for enzyme extractions, evaluating both antioxidant and detoxification systems through the assay of glutathione reductase (GR), guaiacol peroxidase (POD), glutathione peroxidase (GPx), catalase (CAT) as well as glutathione S-transferase (GST). Additionally, thiobarbaturic acid (TBA) method was used to evaluate the peroxidation of lipids (LPO). GST was inhibited in all studied organs at most MC RR concentrations used. Activities of GR, POD and GPx were enhanced in liver at 2 microg L(-1), but inhibited in gills at all tested concentrations. CAT activity was enhanced in liver at all studied concentrations. Antioxidant response in liver is activated at low toxin concentrations, followed by a drop at the highest MC-RR levels. On the contrary, detoxification activity is inhibited in liver and brain in a dose-dependent way. On the other hand, MC-RR (>or=2 microg L(-1)) induced LPO in brain of exposed fish, but not in other organs. This finding becomes to this organ in one of the most severely affected. Results show that gills are also very affected, since both antioxidant and detoxification systems were inhibited in this tissue. Thus, inhibition of these defense systems could increase the uptake of different toxics through gills of fish exposed to dissolved MC-RR, leading to an increased health risk for fish. The different response observed on diverse organs exposed to MC-RR might be related to the uptake route as well as on biotransformation and bioaccumulation capabilities of different studied organs. PMID- 16263187 TI - Ability of Lactobacillus gasseri K 7 to inhibit Escherichia coli adhesion in vitro on Caco-2 cells and ex vivo on pigs' jejunal tissue. AB - The ability of Lactobacillus (Lb.) gasseri K 7 to inhibit adhesion of Escherichia coli O8:K88 to intestinal mucosa was studied on cultured Caco-2 cells and ex vivo on pigs' small intestinal tissue. Lactobacilli were added simultaneously with E. coli, before E. coli and after E. coli for competition, exclusion and displacement assays. The concentration of lactobacilli on fully differentiated Caco-2 cells was 4.5+/-0.3 x 10(8) cfu/well, while the concentration of E. coli varied from 1.5 x 10(6) to 4.3 x 10(8) cfu/well. The number of E. coli adhered to Caco-2 monolayer (cfu/well) was lineary correlated (R(2)=0.97) to the concentration of added cells. In the assay simulating exclusion, E. coli adhesion was reduced by Lb. gasseri K 7 strain by 0.1 to 0.6 log cfu/well. The binding of E. coli was inhibited even more when incubated simultaneously with lactobacilli, particularly at the lowest concentration of E. coli (ratio E. coli/lactobacilli 1:248), where five-times reduction (or 0.7 log) was observed. When adhesion to tissue derived from pigs' jejunum was tested, concentration of E. coli was constant (6.9+/-0.14 x 10(7) cfu/ml), while the concentration of Lb. gasseri K 7 was 5.9 x 10(7) and 1.3 x 10(7) cfu/ml in two independent experiments, respectively. The adhesion of E. coli and Lb. gasseri K 7 cells to jejunal mucosa was similar (1.0+/-0.17 x 10(6) and 1.54+/-0.10 x 10(6) cfu/cm(2)) when the concentrations of single strains in suspensions were approximately the same. No significant competition, exclusion or displacement of E. coli by lactobacilli was observed on jejunal tissue. In conclusion, Lb. gasseri K 7 was found to be effective in reducing E. coli adhesion to Caco-2 enterocytes, but it was not able to do so in ex vivo conditions tested for pig jejunal tissue. PMID- 16263188 TI - An evaluation of sampling- and culturing methods in the Norwegian action plan against Campylobacter in broilers. AB - The Norwegian Action Plan against Campylobacter in broilers was implemented in May 2001 with the objective of reducing human exposure to Campylobacter through Norwegian broilers. From each flock, samples collected at the farm about one week prior to slaughter, and then again at the slaughter plant, are examined for the presence of Campylobacter. All farmers with positive flocks are followed up with bio-security advice. Sampling of broiler products at retail level is also included in the Action Plan. The aim of this study was to evaluate the existing sampling and culturing methods of the Norwegian Action Plan against Campylobacter in broilers. The material collected was pooled faecal samples, pooled cloacae samples and caecae samples from individuals. The highest number of positives, from culturing of the pooled faecal samples, the pooled cloacae swabs and the caecae swabs from individuals, were obtained at incubation temperature 41.5 degrees C. When comparing the results at incubation temperature 37 and 41.5 degrees C, the faecal samples from the farms demonstrated a high concordance, with a kappa value of 0.88. The results from culturing cloacae swabs and caecae samples from slaughter plant level at two temperatures did not agree very well with a kappa value of 0.21 and moderate value of 0.57, respectively, but were both disconcordant at a level of 0.05. Modelling farm level data indicated that if increasing the number of pooled samples per flock from two (in existing regime) to three, the flock sensitivity increases from 89% to 95%. Modelling of slaughter plant data indicated that three pooled cloacae swabs are needed to identify 90% of the positive flocks. The results from the modelling of caecae data indicated that samples from seven individuals are sufficient to identify 90% of the positive flocks and caecae samples could thus be an alternative to cloacae sampling at slaughter plant level. PMID- 16263190 TI - Recombinant polymers for cancer gene therapy: a minireview. AB - A major challenge for successful cancer gene therapy is the development of safe and effective gene delivery vectors. Gene delivery vectors can be viral or nonviral. Among nonviral vectors various polymeric vectors have shown potential in gene delivery. However, much work needs to be done in order to correlate polymer structure with gene release at the target site and transfection efficiency. This article is a brief introduction into cancer gene therapy, barriers and methods for gene transfer with emphasis on the applications of recombinant polymers for cancer gene therapy. PMID- 16263189 TI - Folate-conjugated methoxy poly(ethylene glycol)/poly(epsilon-caprolactone) amphiphilic block copolymeric micelles for tumor-targeted drug delivery. AB - Amphiphilic block copolymers composed of methoxy poly(ethylene glycol) (MPEG) and poly(epsilon-caprolactone) (PCL) were synthesized and then conjugated with folic acid to produce a folate-receptor-targeted drug carrier for tumor-specific drug delivery. Folate-conjugated MPEG/PCL micelles containing the anticancer drug paclitaxel were prepared by micelle formation in aqueous medium. The size of the folate-conjugated MPEG/PCL micelles formed was about 50-130 nm, depending on the molecular weight of block copolymers, and was maintained at less than 150 nm even after loading with paclitaxel. The in vitro release profile of the paclitaxel from the MPEG/PCL micelles exhibited no initial burst release and showed sustained release. Paclitaxel-loaded folate-conjugated MPEG/PCL micelles (PFOL50) exhibited much higher cytotoxicity for cancer cells, such as MCF-7 and HeLa cells, than MPEG/PCL micelles without the folate group (PMEP50). Confocal image analysis revealed that fluorescent paclitaxel-loaded PFOL50 micelles were endocytosed into MCF-7 cells through the interaction with overexpressed folate receptors on the surface of the cancer cells. PMID- 16263191 TI - Ill health and its potential influence on household consumptions in rural China. AB - Ill health is very expensive and could have significant impact on household consumptions. The purpose of this study is to examine the differences in household consumption patterns among households with or without ill health family member(s) in rural China. We also examine the opportunity cost of ill health by estimating the marginal effects of medical spending on consumption patterns. The data used in this study are from the baseline survey of a community-based rural health insurance study in a poor rural area of China conducted in 2002. The unit of analysis in this study is the household; 4553 households are included in this survey. Fractional Logit model is used as our prediction model. Ill health is measured by the presence of hospitalization and presence of diagnosed chronic disease(s) in a household. Findings from this study reveal that ill health and medical expenditure reduces household investment in human capital, physical capital for farm production, and other consumptions that are critical to human well-being. Subgroup analysis displayed that the impacts of medical expenditure on household consumption patterns described above are more significant in low income households than in high-income households. In addition, the decline of the percentages of other consumptions is much larger for households with hospitalization than for households with chronic diseases. PMID- 16263193 TI - Intraocular sustained drug delivery using implantable polymeric devices. AB - Vitreoretinal diseases involving age-related macular degeneration (AMD) are refractory to most topical or systemic drugs. The retina and the vitreous cavity have a unique position regarding pharmacokinetics in that the inner and outer blood retinal barriers separate the retina and vitreous from the systemic circulation. Eye drops achieve minimal therapeutic concentrations in the vitreoretinal tissue. Drug delivery systems are a strategy to address this. Intraocular sustained drug release using implantable devices has been investigated to treat vitreoretinal diseases. Possible targeted diseases include those in which repeated intraocular injections are effective (cytomegalovirus retinitis, uveitis), diseases requiring surgery (proliferative vitreoretinopathy), and chronic diseases (AMD, macular edema, retinitis pigmentosa). Hydrophobic or hydrophilic polymers shaped into a sheet, disc, rod, plug, or a larger device can be implanted into the subretinal space, intrascleral space, vitreous space, peribulbar space, or at the pars plana. Many researchers suggest the feasibility of these implants to treat AMD. PMID- 16263195 TI - Synthesis of new hexahydro- and octahydropyrido[1,2-c]pyrimidine derivatives with an arylpiperazine moiety as ligands for 5-HT1A and 5-HT2A receptors. Part 4. AB - New 4-aryl-2H-pyrido[1,2-c]pyrimidine-1,3-dione derivatives of arylpiperazine (6 18) were prepared and evaluated in vitro for their affinity for 5-HT1A, 5-HT2A, and alpha1 receptors. The influence of ortho substitution in the phenyl ring, substitution at position 4 of the pyrido[1,2-c]pyrimidine system, and its unsaturation degree were explored. The tested compounds showed high affinity for the 5-HT1A receptor (Ki = 1.3-79.2 nM) and moderate to low affinity for the 5 HT2A (Ki = 51.7-1405 nM) and alpha1 receptors (Ki = 19.7-382.3 nM). Compounds 8 and 10 showed the highest 5-HT1A receptor affinity (Ki = 1.3 and 2.2 nM, respectively) and were 37- and 35.9-fold, respectively, more selective in relation to alpha1 adrenoreceptors. PMID- 16263194 TI - Intracellular oxidative activity and respiratory burst of leukocytes isolated from multiple sclerosis patients. AB - Oxidative damage induced by free radicals and reactive oxygen species (ROS) have been suggested to play an important role in the development of autoimmune diseases such as multiple sclerosis (MS) disease and it has been hypothesised that oxidative injury could mediate demyelination and axonal injury in MS subjects. In our study, we compared intracellular oxidative activity and the respiratory burst activity in MS patients (n=20) and healthy controls (n=15) using leukocytes as cellular model. At this purpose, intracellular ROS levels were evaluated by fluorometric assay using the 2'-7'-dichlorodihydrofluorescin diacetate probe (H(2)DCFDA) in untreated or in leukocytes stimulated with phorbol 12-myristate-13-acetate (PMA). Our results demonstrate that the intracellular spontaneous ROS production in leukocytes from MS patients was higher with respect to cells from control subjects (p<0.001). PMA addition induced a higher formation of ROS both in leukocytes from MS patients and controls (p<0.001). The PMA induced production of ROS was significantly higher in leukocytes from MS with respect to controls (p<0.001). Significant positive correlations were established between intracellular spontaneous or PMA-induced production of ROS in leukocytes isolated from MS patients and the clinical parameters used to evaluate disease disability such as expanded disability status scale (EDSS), brain lesions evaluated by MRI and visual evoked potential (VEP) (p<0.001). In conclusion, our results demonstrate higher levels of intracellular ROS in untreated or in PMA treated leukocytes isolated from MS patients with respect to healthy subjects confirming the role of oxidative stress in multiple sclerosis. PMID- 16263196 TI - Evaluation of osteotomy fixation changing the number, the extension and the location of the plates. AB - We used a two-dimensional model that simulated a fractured human mandible to study the extent of load resistance of different plates. We studied 49 bovine ribs in seven groups using varying number (1 or 2), lengths of plates (4 or 6 holes), and location (near the lower or upper border). The number and location of plates had a greater effect on resistance to load than length of plates. The best mean resistance values were achieved by a combination of two parallel plates secured near the lower and upper borders, followed by single plates secured near the upper border. The lowest values were with single plates secured close to the lower border. PMID- 16263197 TI - Partial avulsion of the inferior rectus: an unusual cause of diplopia after blunt orbital trauma. AB - A 14-year-old boy had a severe blow to the right orbit, which caused a blow-out fracture of the orbital floor and damage to the inferior rectus with resultant diplopia. After two operations his vision is still not perfect. PMID- 16263198 TI - Re: Shinohara E H, Takahashi A. View of the epiglottis during examination of the oral cavity. Br J Oral Maxillofac Surg 2005;43:264-5. PMID- 16263199 TI - Scaphoid non-union: the role of vascularized grafting in recalcitrant non-unions of the scaphoid. AB - Achieving union using conventional grafts has a high chance of failure in patients with recalcitrant non-union (persistent pseudarthrosis) of the scaphoid bone, an avascular proximal fragment and previous failed surgeries because of poor host bed vascularity. Eleven patients with long-standing non-union were treated with vascularized pedicle bone grafting and supplementary corticocancellous grafting. Five had screw fixation and six were fixed with K wires. The average age of the patients was 28 years, average duration of the non union was 39 months and mean radiological follow-up was 32 months. There were no significant skeletal complications, although two patients developed neuromata. At review, only six of the 11 non-unions were united. Whilst this is a difficult clinical problem and achieving union is a formidable challenge, we believe that there is a role for such extensive surgery in order to achieve good postoperative function. PMID- 16263200 TI - Eisenia fetida increased removal of polycyclic aromatic hydrocarbons from soil. AB - The removal of phenanthrene, anthracene and benzo(a)pyrene added at three different concentrations was investigated with or without earthworms (Eisenia fetida) within 11 weeks. Average anthracene removal by the autochthonous micro organisms was 23%, 77% for phenanthrene and 13% for benzo(a)pyrene, while it was 51% for anthracene, 47% for benzo(a)pyrene and 100% for phenanthrene in soil with earthworms. At 50 and 100mg phenanthrene kg(-1)E. fetida survival was 91% and 83%, but at 150 mg kg(-1) all died within 15 days. Survival of E. fetida in soil amended with anthracene < or = 1000 mg kg(-1) and benzo(a)pyrene < or = 150 mg kg(-1) was higher than 80% and without weight loss compared to the untreated soil. Only small amounts of PAHs were detected in the earthworms. It was concluded that E. fetida has the potential to remove large amounts of PAHs from soil, but more work is necessary to elucidate the mechanisms involved. PMID- 16263201 TI - Ventilator dependence and expressions of need: a study of patients with amyotrophic lateral sclerosis in Japan. AB - This research examined the experience of Amyotrophic Lateral Sclerosis (ALS) patients who depend on an invasive mechanical ventilator in Japan. We investigated their difficulties in expressing their needs, their desires and wishes, and their sources of support and happiness. We examined the relationship between these factors and patient demographics, and hope as an indicator of successful adaptation to the illness experience (assessed by the Herth Hope Index). Interview results guided the formation of an anonymous questionnaire distributed to patients by mail. We interviewed 27 patients and their families and surveyed 157 respondents with a questionnaire. Most patients experienced multiple categories of difficulties, which correlated with reduced hope. More severe physical symptoms correlated with more emotional and social difficulties. Notable findings included a high prevalence of unalleviated pain, fear or experience of ventilator difficulties, and fear of burdening others. Having more sources of psychosocial support and happiness was associated with greater hope. Living at home was associated with fewer social difficulties. No patients claimed additional sources of support without claiming family or professional caregiver support, suggesting their mediation may be crucial in maintaining other social connections. Users of computer communication reported more sources of support and happiness and less frustration from difficulty expressing themselves. The most common reported desires, following a cure for ALS, related to the happiness of the patients' families, and a desire not to burden them. We also found that invasive mechanical ventilation (IMV) had been initiated emergently in 30.1% of patients without patient or family consent. Our results provide an insight into the world of this challenged population, elucidating the difficulties they face, and clarifying the role of support and other factors in maintaining hope. We identify concrete areas to which increased attention should be directed in patient care. PMID- 16263202 TI - Influence of orange juice in the levels and in the genotoxicity of iron and copper. AB - World consumption of natural juices is increasing as a consequence of the human search for a healthier life. The juice production industry, especially for orange juice, is expanding in several countries and particularly in Brazil. Despite scientific data reporting beneficial properties derived from juice consumption, some components of juices have been identified as mutagenic or carcinogenic. Carcinogenic or genotoxic effects may be mediated by the interaction of juice components with transition metals or by sub-products of juice auto-oxidation. In this study, the mutagenic potential of orange juice and two metallic agents used in dietary supplementation, FeSO(4) and CuSO(4), were investigated using the comet assay in mouse blood cells (in vivo). Both metal compounds were genotoxic for eukaryotic cells after 24h treatment at the doses used. Significant damage repair was observed after 48h of treatment with the same compounds. Orange juice had a modulating effect on the action of metallic sulfates. In the case of iron treatment, the presence of the orange juice had a preventive, but not restorative, effect. On the other hand, in the case of copper treatment, the effects were both preventive and restorative. PIXE (particle induced X-ray emission) analysis indicated a positive correlation between DNA damage and the hepatic levels of iron and a negative correlation between whole blood copper and DNA damage. A negative correlation between hepatic iron and whole blood copper content was also seen in the treatment with both ferrous and cupric sulfates. PMID- 16263203 TI - Cytometric analysis of lidocaine-induced cytotoxicity: a model experiment using rat thymocytes. AB - Lidocaine is a local anesthetic possessing both lipophilic and hydrophilic properties. It also acts as a surfactant. Thus, the disruption of membranes, resulting in necrosis, is one of possible mechanisms for lidocaine-induced cytotoxicity. However, lidocaine is reported to induce apoptosis. Therefore, in order to compare two mechanisms for cell death induced by lidocaine, the effects of millimolar lidocaine were examined on rat thymocytes by a flow cytometer with appropriate fluorescent probes. Lidocaine decreased the population of living cells with phosphatidylserine-exposed membranes, one of markers for early stage of apoptosis, and increased the population of dead cells without increasing that of cells with hypodiploidal DNA. Lidocaine at millimolar concentrations may deteriorate the membranes of such apoptotic living cells rather than those of intact living cells, resulting in necrosis. It is suggested that the process of apoptosis is not completed in the presence of millimolar lidocaine. PMID- 16263204 TI - Intrinsic membrane properties of vertebrate vestibular neurons: function, development and plasticity. AB - Central vestibular neurons play an important role in the processing of body motion-related multisensory signals and their transformation into motor commands for gaze and posture control. Over recent years, medial vestibular nucleus (MVN) neurons and to a lesser extent other vestibular neurons have been extensively studied in vivo and in vitro, in a range of species. These studies have begun to reveal how their intrinsic electrophysiological properties may relate to their response patterns, discharge dynamics and computational capabilities. In vitro studies indicate that MVN neurons are of two major subtypes (A and B), which differ in their spike shape and after-hyperpolarizations. This reflects differences in particular K(+) conductances present in the two subtypes, which also affect their response dynamics with type A cells having relatively low frequency dynamics (resembling "tonic" MVN cells in vivo) and type B cells having relatively high-frequency dynamics (resembling "kinetic" cells in vivo). The presence of more than one functional subtype of vestibular neuron seems to be a ubiquitous feature since vestibular neurons in the chick and frog also subdivide into populations with different, analogous electrophysiological properties. The ratio of type A to type B neurons appears to be plastic, and may be determined by the signal processing requirements of the vestibular system, which are species variant. The membrane properties and discharge pattern of type A and type B MVN neurons develop largely post-natally, through the expression of the underlying ion channel conductances. The membrane properties of MVN neurons show rapid and long-lasting plastic changes after deafferentation (unilateral labyrinthectomy), which may serve to maintain their level of activity and excitability after the loss of afferent inputs. PMID- 16263205 TI - Nuclear magnetic resonance and thermal studies on the interaction between salicylic acid and model membranes. AB - DSC and (1H and 31P) NMR measurements are used to investigate the perturbation caused by the keratolytic drug, salicylic acid (SA) on the physicochemical properties of the model membranes. Model membranes (in unilamellar vesicular (ULV) form) in the present studies are prepared with the phospholipids, dipalmitoyl phosphatidylcholine (DPPC), dipalmitoyl phosphatidylethanolamine (DPPE), dipalmitoyl phosphatidic acid (DPPA) and mixed lipid DPPC-DPPE (with weight ratio, 2.5:2.2). These lipids have the same acyl (dipalmitoyl) chains but differed in the headgroup. The molar ratio of the drug to lipid (lipid mixture), is in the range 0 to 0.4. The DSC and NMR results suggest that the lipid head groups have a pivotal role in controlling (i) the behavior of the membranes and (ii) their interactions with SA. In the presence of SA, the main phase transition temperature of (a) DPPE membrane decreases, (b) DPPA membrane increases and (c) DPPC and DPPC-DPPE membranes are not significantly changed. The drug increases the transition enthalpy (i.e., acyl chain order) in DPPC, DPPA and DPPC-DPPE membranes. However, the presence of the drug in DPPC membrane formed using water (instead of buffer), shows a decrease in the transition temperature and enthalpy. In all the systems studied, the drug molecules seem to be located in the interfacial region neighboring the glycerol backbone or polar headgroup. However, in DPPC-water system, the drug seems to penetrate the acyl chain region also. PMID- 16263206 TI - Complexation of daunomycin with a DNA oligomer in the presence of an aromatic vitamin (B2) determined by NMR spectroscopy. AB - The effect of simultaneous binding of the anthracycline antibiotic Daunomycin (DAU) and the Vitamin B2 derivative, Flavin-mononucleotide (FMN), with the DNA oligomer, d(TGCA)2, in solution has been investigated quantitatively by 1I-NMR spectroscopy (500 MHz). The equilibrium reaction constants and the thermodynamical parameters (DeltaH, DeltaS) of the hetero-association FMN-DAU and complexation of FMN with d(TGCA)2 have been determined by analysis of the concentration and temperature dependences of chemical shifts of the aromatic protons in terms of a competitive binding model. A criterion for discrimination between hetero-association and DNA complexation has been developed and applied to the analysis of the simultaneous binding of the antibiotic and the vitamin with DNA. Under the conditions of the experiment, it is found that both the hetero association of FMN with DAU and the complexation of FMN with DNA contribute approximately equally to the decrease of DAU binding with DNA oligomer. Such competitive complexation of aromatic vitamin and drug with DNA could affect the biological activity of such drugs. PMID- 16263207 TI - Evaluating preference trials of oral phosphodiesterase 5 inhibitors for erectile dysfunction. AB - More treatment options are available now for the treatment of erectile dysfunction (ED) than ever. Treatments include oral phosphodiesterase 5 (PDE5) inhibitors, intracavernosal injections, vacuum constriction devices, and penile implants. Clinicians, researchers, and patients are interested in making direct comparisons between the response of newer treatments and that of established and more developed therapies. Of the currently available treatment options for ED, the most commonly prescribed therapies are oral PDE5 inhibitors, which include sildenafil citrate (Viagra, Pfizer Inc), tadalafil (Cialis, Lilly ICOS), and vardenafil (Levitra, Bayer). However, most patient preference studies of these drugs conducted to date have serious design flaws that hinder interpretation of the data, and thus limit the utility of the results. To make an informed decision on the most appropriate treatment option available, physicians and their patients require a thorough understanding of the methodology of these studies. Clinical comparison or preference trials must establish internal and external validity if the data are to be used in a generalized patient population. We review preference studies that compared sildenafil, tadalafil, and vardenafil, and highlight study designs that can introduce bias. We propose that, like safety and efficacy trials, randomized controlled trials (RCTs) should be the gold standard for evaluating patient preference treatments for ED. We do not wish to discourage individual investigators from performing preference studies, but rather to highlight the features of current preference trials to help patients and clinicians alike become aware of potential biases from independent or industry sponsored patient preference trials so that they can interpret the results accordingly. Key components of patient preference RCTs are reviewed: period and carryover effects, preference assessments, eligibility criteria, and data analysis. We discuss why these components of patient-preference RCTs are important for evaluating the validity and relevance of patient preference studies. The preference studies discussed in this brief review are summarized in , and the methodological problems with each study are indicated. We provide a recommendation for the design of such trials that can minimize bias and provide better data for physicians and their patients. PMID- 16263208 TI - Randomized, double-blind, placebo-controlled crossover study in men with prostate cancer and rising PSA: effectiveness of a dietary supplement. AB - OBJECTIVES: Epidemiological studies have shown significant relationships between the use of dietary components and prostate cancer incidence and mortality. Large studies of primary prevention, which confirm these findings, are desirable but costly and difficult to design. The present tertiary prevention study reports on the effect of a dietary supplement in comparison with placebo on the rate of increase of prostate-specific antigen (PSA). METHODS: 49 patients with a history of prostate cancer and rising PSA levels after radical prostatectomy (n = 34) or radiotherapy (n = 15) participated in a randomised, double-blind, placebo controlled crossover study of a dietary supplement. Ethical approval of the protocol was obtained. Treatment periods of 10 weeks were separated by a 4-week washout period. The supplement consisted of soy, isoflavones, lycopene, silymarin and antioxidants as main ingredients. Changes in the rate of increase of PSA (PSA slope and doubling time) were the primary parameters of efficacy. Analyses according to intention to treat (ITT) and per protocol (PP) were carried out. RESULTS: Baseline parameters did not differ between randomised groups. Five participants were lost to follow-up, however 46 could be evaluated in an ITT analysis. PP analysis could be performed in 42 men with at least 5 PSA measurements. Per protocol analysis showed a significant decrease in PSA slope (p = 0.030) and (2)log PSA slope (p = 0.041). This translates into a 2.6 fold increase in the PSA doubling time from 445 to 1150 days for the supplement and placebo periods. No treatment-based changes in safety parameters were observed during the study. CONCLUSIONS: The soy-based dietary supplement utilised in this study was shown to delay PSA progression after potentially curative treatment in a significant fashion. More extensive studies of the supplement may be indicated. PMID- 16263209 TI - Treatment of oil-in-water emulsions: performance of a sawdust bed filter. AB - The effect of operating conditions on the performance of a sawdust bed filter used for the treatment of an oil-in-water emulsion was investigated. A metalworking fluid (3 vol.% oil) was used as oil-in-water emulsion and sawdust as filter medium and sorbent. Because of the high stability of the emulsion, small amounts of inorganic salt (calcium sulphate) were mixed with the sorbent, acting as coagulant to achieve the emulsion breakdown. The influence of flow rate, bed height, temperature and the amount of coagulant salt added was studied. Experimental results show that several processes are involved in oil removal from oil-in-water emulsions, i.e. coagulation, coalescence, adsorption or straining. More than 99% of oil content in the influent stream was removed. Experimental results show that low-cost sorbents like sawdust are feasible to be used in the treatment of oil-in-water emulsions if small amounts of coagulant salts are added to the filter media. PMID- 16263210 TI - Calculation and usage of containment monitor radiation readings during PWR accidents. AB - A computer code named CALCON for calculation of containment monitor radiation readings is introduced. The validity of the code was verified by comparison with data given in IAEA technical documents. The contribution of isotopes to containment readings under conditions of core melt, gap release and normal coolant release were investigated. The conclusions were that the radiation reading in containment is mainly from iodine and noble gases when sprays are off, and the radiation reading is mainly from noble gases when sprays are on, and during the beginning hours when radionuclides are released into containment, the monitor readings will decrease rapidly. Curves of containment radiation readings versus shutdown time for DAYA BAY nuclear power plant were calculated using CALCON. PMID- 16263211 TI - Production of granular activated carbon from waste Rosa canina sp. seeds and its adsorption characteristics for dye. AB - An activated carbon was developed from Rosa canina sp. seeds, characterized and used for the removal of methylene blue (basic dye) from aqueous solutions. Adsorption studies were carried out at 20 degrees C and various initial dye concentrations (20, 40, 60, 80, and 100 mg/L) for different times (15, 30, 60, and 120 min). The adsorption isotherm was obtained from data. The results indicate that the adsorption isotherm of methylene blue is typically S-shaped. The shape of isotherm is believed to reflect three distinct modes of adsorption. In region 1, the adsorption of methylene blue is carried out mainly by ion exchange. In region 2 by polarizations of pi-electrons established at cyclic parts of the previously adsorbed methylene blue molecules is occurred. However, it is not observed any change at the sign of the surface charge although zeta potential value is decreased with increase of amount adsorbed. In region 3, the slope of the isotherm is reduced, because adsorption now must overcome electrostatic repulsion between oncoming ions and the similarly charged solid. Adsorption in this fashion is usually complete when the surface is covered with a monolayer of methylene blue. To reveal the adsorptive characteristics of the produced active carbon, porosity and BET surface area measurements were made. Structural analysis was performed using SEM-EDS. The produced active carbon has the specific surface area of 799.2 m2 g-1 and the iodine number of 495 mg/g. PMID- 16263212 TI - Integrated effects of selected ions on 2,4,6-trinitrotoluene-removal by O3/H2O2. AB - Considering the components of 2,4,6-trinitrotoluene (TNT)-containing water, this paper aims to research the integrated effects of ions on TNT-removal by O(3)/H(2)O(2) through the selection of CO(3)(-), HCOO(-), Cu(2+) and Al(3+). In view of TNT-removal rate and its constant, we find that the test with HCO(3)(-) or HCOO(-) or Cu(2+) results in lower TNT-removal rate and its constant than control test. Therefore, it may conclude that HCO(3)(-) or HCOO(-) and Cu(2+) has a potential to inhibit the efficacy of O(3)/H(2)O(2), and that their inhibitions increase with an order from HCOO(-) to Cu(2+) and to HCO(3)(-). However, Al(3+) is an exception, because it has a potential to improve the efficacy. When the two selected ions coexist, HCO(3)(-) and HCOO(-) inhibit the efficacy. The inhibition is greater than that of either one alone, and also greater than their sum, and thus the integrated effect of HCO(3)(-) and HCOO(-) follows the synergistic effect. The inhibition of Cu(2+) and HCOO(-) coexistence also is greater than that either alone, but smaller than their sum, and thus their integrated effect follows the independent effect. The integrated effect of Al(3+) and HCOO(-) follows the addition effect. PMID- 16263213 TI - Bench-scale column experiments to study the containment of Cr(VI) in confined aquifers by bio-transformation. AB - Bench-scale soil column experiments were conducted to study the effectiveness of Cr(VI) containment in confined aquifers using in situ bio-transformation. Batch adsorption studies were carried out to estimate the adsorption capacities of two different soils for Cr(VI) and Cr(III). Bio-kinetic parameters were evaluated for the enriched microbial system. The inhibition constant, evaluated using Monod's inhibition model, was found to be 11.46 mg/L of Cr(VI). Transport studies indicated that it would not be possible to contain Cr(VI) by adsorption alone. Transport and bio-transformation studies indicated that the pore velocity and the initial bio-mass concentration significantly affect the containment process. In situ bio-remediation is effective in the case of silty aquifers. Cr(VI) concentration of 25 mg/L was effectively contained within 60 cm of a confined silty aquifer. Cr(VI) containment could be achieved in sandy aquifers when the pore velocity was very low and the initial augmented bio-mass was high. A bio barrier of approximately one meter width would be able to contain Cr(VI) if the initial Cr(VI) concentration is as much as 25 mg/L. PMID- 16263214 TI - Choroid plexus epithelial cells co-express the long and short form of the leptin receptor. AB - Two types of leptin receptors (Ob-R) are structurally and functionally characterized. The Ob-Ra, also called short form of leptin receptor, and the Ob Rb, which is functionally coupled to intracellular signalling pathways. In the CNS, the Ob-Ra is mainly present in the choroid plexus and in the blood-brain barrier and it is involved in leptin transport from the periphery to the brain. The Ob-Rb is widely distributed in the hypothalamus and in many peripheral tissues and is coupled to the JAK/STAT signalling pathway. In this work, we describe the presence of both Ob-Ra and Ob-Rb in a cell line of choroid plexus epithelium. In addition, we demonstrate that Ob-Rb in the choroid plexus is functionally coupled to intracellular signalling pathways as leptin induced the phosphorylation of the signal transducer and activator of transduction 3 (STAT3). PMID- 16263215 TI - Platelet activating factor and its metabolite promote sleep in rabbits. AB - Platelet activating factor (PAF) is a key inflammatory mediator. PAF and its receptor are found in brain and PAF affects or is affected by the production of sleep promoting cytokines such as interleukin-1. PAF also interacts with several other sleep-regulatory substances such as nerve growth factor, brain-derived neurotrophic factor, neurotrophin-3, nitric oxide, prostaglandins, and prolactin. We thus hypothesized that PAF would increase sleep. In these experiments, each rabbit received an injection of 25 microl of 2% DMSO to obtain control values, and on a separate day received PAF or lyso-PAF, a metabolite of PAF. Ten, 100 and 500 nmol for each substance was injected intracerebroventricularly. Both PAF and lyso-PAF enhanced non-rapid eye movement (NREM) sleep but not REM sleep. Lyso PAF, but not PAF, induced hyperthermia. Results are consistent with the hypothesis that the brain cytokine network is involved in physiological sleep regulation. PMID- 16263216 TI - Detection of cerebral oxyhaemoglobin changes during vestibular Coriolis cross coupling stimulation using near infrared spectroscopy. AB - Near infrared spectroscopy (NIRS) has been successful in monitoring cerebral haemodynamics when the subject is immobilized during surgery, and when there is a drastic depletion of blood from the cerebral cortex during positive acceleration. In this study, we monitored subtle changes of cerebral oxygen level using NIRS during vestibular stimulation. For the control conditions, cerebral oxygen status was monitored in six stationary subjects sitting upright, and while they executed head movements in the pitch axis with eyes opened and eyes closed. The experimental conditions involved the subjects making a head movement which required a 45 degrees pitch-down followed by a return to upright head movements 12 s later during yaw rotation (Coriolis cross coupling) at 10 and 20 rotations per minute (rpm) in a random order. Oxyhaemoglobin (O(2)Hb), deoxyhaemoglobin (HHb) and total haemoglobin levels were recorded every 0.5 s from both the parietal and the occipital lobe simultaneously. A significant rotation effect was observed in total Hb level changes from baseline in both regions. Occipital O(2)Hb increased significantly after the head movement with eyes opened at 20 rpm. Our findings appear to be consistent with previous vestibular studies that significant changes in brain blood flow occur during caloric stimulation. NIRS can be used to monitor discrete cortical blood flow changes resulting from vestibular and other forms of stimulation. PMID- 16263217 TI - Event-related brain potential variations during location and identity negative priming. AB - Event-related brain potential (ERP) correlates of the negative priming (NP) effect were studied in this experiment. One task required responses to targets which were or were not distractors in the preceding trial (identity priming); in the other task, the location of the distractor in the preceding trial was or was not the same as the location of the actual target (location priming). Whereas both tasks were associated with reaction time increases for negative priming, we obtained differential effects for ERP data. The parieto-occipital P1-N1 amplitude was reduced and P3 latency was delayed in response to location NP targets. No such effects were found for identity NP. The P3 amplitude was significantly larger after identity NP but not after location NP targets. It was concluded that location NP appears to be related to the early inhibition of sensory processing and to a slowing of the stimulus evaluation process. No ERP indications of inhibitory mechanisms were found with the identity NP task. The larger P3 amplitudes in the identity priming task may indicate increased attentional resources that are necessary for the processing of NP targets. PMID- 16263218 TI - Do eosinophils have a role in the severity of Babesia annae infection? PMID- 16263219 TI - Antiangiogenic peptides and proteins: from experimental tools to clinical drugs. AB - The formation of a 'tumor-associated vasculature', a process referred to as tumor angiogenesis, is a stromal reaction essential for tumor progression. Inhibition of tumor angiogenesis suppresses tumor growth in many experimental models, thereby indicating that tumor-associated vasculature may be a relevant target to inhibit tumor progression. Among the antiangiogenic molecules reported to date many are peptides and proteins. They include cytokines, chemokines, antibodies to vascular growth factors and growth factor receptors, soluble receptors, fragments derived from extracellular matrix proteins and small synthetic peptides. The polypeptide tumor necrosis factor (TNF, Beromun) was the first drug registered for the regional treatment of human cancer, whose mechanisms of action involved selective disruption of the tumor vasculature. More recently, bevacizumab (Avastin), an antibody against vascular endothelial growth factor (VEGF)-A, was approved as the first systemic antiangiogenic drug that had a significant impact on the survival of patients with advanced colorectal cancer, in combination with chemotherapy. Several additional peptides and antibodies with antiangiogenic activity are currently tested in clinical trials for their therapeutic efficacy. Thus, peptides, polypeptides and antibodies are emerging as leading molecules among the plethora of compounds with antiangiogenic activity. In this article, we will review some of these molecules and discuss their mechanism of action and their potential therapeutic use as anticancer agents in humans. PMID- 16263221 TI - Does meditation predispose to epilepsy? EEG studies of expert meditators self inducing simple partial seizures. PMID- 16263222 TI - Morphological evidence of marine adaptations in human kidneys. AB - Amongst primates, kidneys normally exhibiting lobulated, multipyramidal, medullas is a unique attribute of the human species. Although, kidneys naturally multipyramidal in their medullary morphology are rare in terrestrial mammals, kidneys with lobulated medullas do occur in: elephants, bears, rhinoceroses, bison, cattle, pigs, and the okapi. However, kidneys characterized with multipyramidal medullas are common in aquatic mammals and are nearly universal in marine mammals. To avoid the deleterious effects of saline water dehydration, marine mammals have adaptively thickened the medullas of their kidneys--which enhances their ability to concentrate excretory salts in the urine. However, the lobulation of the kidney's medullary region in marine mammals appears to be an adaptation to expand the surface area between the medulla and the enveloping outer cortex in order to increase the volume of marine dietary induced hypertonic plasma that can be immediately processed for the excretion of excess salts and nitrogenous waste. A phylogenetic review of freshwater aquatic mammals suggest that most, if not all, nonmarine aquatic mammals inherited the medullary pyramids of their kidneys from ancestors who originally inhabited, or frequented, marine environments. So this suggest that most, if not all, aquatic mammals exhibiting kidneys with lobulated medullas are either marine adapted--or are descended from marine antecedents. Additionally, a phylogenetic review of nonhuman terrestrial mammals possessing kidneys with multipyramidal medullas suggest that bears, elephants and possibly rhinoceroses, also, inherited their lobulated medullas from semiaquatic marine ancestors. The fact that several terrestrial mammalian species of semiaquatic marine ancestry exhibit kidneys with multipyramidal medullas, may suggest that humans could have, also, inherited the lobulated medullas of their kidneys from coastal marine ancestors. And a specialized marine diet in ancient human ancestry could, also, explain the reactivation and enumeration of corporeal eccrine sweat glands and the copious secretion of salt tears. The substantial loss of genetic variation in humans relative to other hominoid primates, combined with the apparent isolation of early Pliocene human ancestors from particular retroviruses that infected all other African primate species, may suggest that such a semiaquatic marine phase, during the emergence of Homo, may have occurred on an island off the coast of Africa during the early Pliocene. PMID- 16263220 TI - Regulation of cocaine-induced activator protein 1 transcription factors by the extracellular signal-regulated kinase pathway. AB - Extracellular signal-regulated kinases and activator protein 1 transcription factor have been functionally linked to addiction. It has also been shown that extracellular signal-regulated kinase activation can regulate cocaine-induced expression of c-Fos and FosB, two possible components of activator protein 1. A direct link between extracellular signal-regulated kinases and activator protein 1 activation has, however, remained unexplored. In this study, we investigated the role of extracellular signal-regulated kinases in the regulation of DNA binding activity and composition of activator protein 1 induced in the mouse caudate putamen by cocaine treatment. We have found that pre-treatment with SL327, a selective inhibitor the extracellular signal-regulated kinase pathway, has no influence on cocaine-induced DNA-binding activity of activator protein 1, when examined one hour after an acute cocaine treatment. This phenomenon results from simultaneous decrease of c-Fos protein level and increases in JunB and deltaFosB protein levels. SL327 pre-treatment, however, reduces the DNA-binding activity of the activator protein 1 complex induced six hours after an acute cocaine treatment as well as one hour after the last of the chronic cocaine injections, a phenomenon that results from the concomitant reduction of all cocaine-induced proteins (c-Fos, FosB, deltaFosB, JunB). In conclusion, we have found that extracellular signal-regulated kinase inhibition may not only interfere with cocaine-induced gene expression and activator protein 1 complex activation, but may also disturb the time-course of gene expression and composition of activator protein 1 complex. Our results support the notion that inhibitors of the extracellular signal-regulated kinase pathway could be valuable tools to obliterate cocaine-induced molecular changes and the development of addiction. PMID- 16263223 TI - FcRY, an Fc receptor related gene differentially expressed during B lymphocyte development and activation. AB - A bioinformatics approach has lead to the identification of FcRY, a new Fc receptor related gene. FcRY is predicted to encode a protein with three immunoglobulin (Ig) domains followed by a mucin-like domain containing a proline rich stalk and a C-terminal leucine rich region. The predicted protein lacks a hydrophobic domain for insertion into the plasma membrane, suggesting that FcRY is an intracellular or secreted protein. This feature is shared with the product of the FcRX/FCRL/FREB gene that is closely linked to FcRY in both human and mouse genomes. Fcry transcripts are first detectable among mouse B lineage cells at the pre-B cell stage. Splenic B cells of the newly formed, follicular, and marginal zone subsets express Fcry, as do germinal center B cells to a lesser extent. FcRY is also expressed in subpopulations of human B cells. A consistent characteristic of FcRY in both species is low level gene expression, which can be further downregulated in normal mouse B cells by signaling through the B cell receptor (BCR) or CD40, thereby suggesting a correlation between cell cycle entrance and diminished FcRY expression. Fcry is upregulated by short-term treatment with BAFF/BLyS, which promotes B cell survival rather than proliferation. LPS induces very rapid but transient enhancement. We observed a pronounced upregulation of Fcry expression in WEHI 231 cells induced by BCR crosslinking to undergo cell cycle arrest prior to apoptosis, consistent with the possible regulation of Fcry expression by cell cycle status. PMID- 16263224 TI - Bovine viral diarrhoea virus (BVDV) subgenotypes in diagnostic laboratory accessions: distribution of BVDV1a, 1b, and 2a subgenotypes. AB - The prevalence of bovine viral diarrhoea virus (BVDV) biotypes and subgenotypes was determined from 131 BVDV positive samples from a diagnostic laboratory. The majority of the isolates were from Oklahoma; however, other states including Kansas, Texas, and Arkansas were represented. These BVDV samples were from submissions of 76 live animals and 55 necropsy samples. There were 131 BVDV samples represented by 117 noncytopathic (NCP), 11 cytopathic (CP) and 3 cases with mixed NCP and CP biotypes. The NCP isolates were more common (P < 0.05) than the CP and NCP/CP combination. The BVDV samples were segregated into three subgenotypes by differential PCR and sequencing of a viral genomic region, 5' untranslated region (5'-UTR). There were more BVDV1b subgenotypes 60/131 (45.8%) than BVDV1a, 37/131 (28.2%) or BVDV2a, 34/131 (26.0%) (P < 0.05). The organ system involvement included the major categories such as respiratory, digestive, mixed/multiple organs, abortions, and persistent infections (PI). All three BVDV subgenotypes were found in persistently infected (PI) cattle and respiratory diseases, both major requests for BVDV diagnosis. Only one of the 131 viruses was genetically similar to the strains present in U.S. vaccines. PMID- 16263225 TI - The management of congenital talipes equinovarus. AB - Congenital talipes equinovarus is a common deformity that is present at birth. It can be treated conservatively. Of the techniques available, the Ponseti method is effective in correcting most of these foot deformities and is best started early. Some of the stiffer "teratological" foot deformities may require surgical releases. Other conservative methods may be applied to correct deformity, but they have not been demonstrated to be as effective as the Ponseti method, although they may reduce the extent of subsequent surgical releases. Surgery can be undertaken to correct deformity, but some loss of motion is inevitable with extensive open procedures. Complications of treatment can be divided into: failure to correct, recurrence and overcorrection. Outcome can be assessed clinically and radiologically. Radiological outcome does not necessarily correspond to clinical results. The best long-term clinical outcome has been reported using the Ponseti method, for which there is great and increasing demand, largely driven by parents and facilitated by the Internet. PMID- 16263226 TI - Differential effects of PCBs on the induction of apoptosis machinery and PKCalpha translocation in rat renal tubular cell cultures. AB - We have demonstrated previously [Perez-Reyes, P.L., Sanchez-Alonso, J.A., Lopez Aparicio, P., Recio, M.N., Perez-Albarsanz, M.A., 2001. Different molecular capacity in the induction of apoptosis by polychlorinated biphenyl congeners in rat renal tubular cell cultures. Biosci. Rep. 6, 765-778] that the polychlorinated biphenyls (PCBs) cause loss of cell viability and accelerate apoptosis in cell kidney cultures. Further investigations are necessary to elucidate the mechanism of apoptosis induction. In this way, we have analyzed in the present work the effects of PCBs on protein kinase C (PKC, a protein family intimately involved in the regulation of cell survival) and the expression of two proapoptotic (caspase-3 and Bax) and one antiapoptotic (Bcl-2) proteins. Aroclor 1248 (a commercial PCB mixture with 48% chlorine by weight), PCB 153 (2,2',4,4',5,5'-hexachlorobiphenyl, a di-ortho-substituted nonplanar congener) and PCB 77 (3,3',4,4'-tetrachlorobiphenyl, a non-ortho-substituted planar congener), significantly increased PKCalpha activity compared to control cells in the cytosolic and particulate cell fractions, and increased the PKCalpha protein content in the particulate fraction. The nonplanar PCB 153 showed stronger effects than the coplanar congener PCB 77. In addition, Aroclor 1248 decreased both, procaspase-3 levels and the Bcl-2/Bax protein ratio. These findings indicate that PCBs, particularly nonplanar congeners, can induce apoptosis in primary renal tubular cells through the PKCalpha, caspase-3 and Bcl-2/Bax pathway. PMID- 16263227 TI - Assessment of soil remediation workers' exposure to polycyclic aromatic hydrocarbons (PAH): biomonitoring of naphthols, phenanthrols, and 1-hydroxypyrene in urine. AB - Urinalysis of multiple polycyclic aromatic hydrocarbons (PAH) biomarkers has been applied to assess the exposure of soil remediation workers on a former creosote wood impregnation site polluted with creosote oil. The uptake of PAHs was measured in preshift, end-of-shift, evening, and next preshift specimens (n=33) of nine volunteers with diverse tasks, using sensitive HPLC-FD methods. The ranges of biomarker concentrations in urine (nmol/l) were: 1-naphthol (14-159), 2 naphthol (9-166), 1- plus 2-naphthol (35-269), 1-hydroxyphenanthrene (OHPhe) (6 56), 2- plus 3-OHPhe (6-70), 4-OHPhe (1-6), 9-OHPhe (1-7), the sum of phenanthrols (15-135), and 1-hydroxypyrene, OHP (2.2-67). Eight of nine workers had OHP levels higher than the Finnish biological limit value for non occupationally exposed persons (3nmol/l). A linear correlation was observed between 1- and 2-naphthol (r=0.90). The biomarker OHP correlated well in urine both with the major (1-OHPhe, r=0.96; 2- plus 3-OHPhe, r=0.84) and the minor phenanthrene metabolites (4-OHPhe, r=0.77; 9-OHPhe, r=0.68), and with the sum of all phenanthrols (r=0.94), but not so well with the sum of naphthols (r=0.66, p<0.001). The smokers had 2.9-, 2.2-, and 4.8-fold higher average concentrations of naphthols, phenanthrols, and OHP, respectively, than the non-smokers. The PAH biomarker data (concentrations and diurnal excretion profiles) showed significant work-related exposure in both non-smoking and smoking subjects. The average exposure levels were clearly higher than those we have measured for instance in asphalt paving workers. The workers' exposure should be assessed by biological monitoring, because at this type of outdoor work the dermal and pulmonary uptake of PAHs are both likely. Adequate measures for preventing, particularly, dermal absorption are of crucial importance for reducing the workers' risk of exposure to carcinogens on soil remediation sites. PMID- 16263228 TI - Tumor necrosis factor alpha partially contributes to lipopolysaccharide-induced intra-uterine fetal growth restriction and skeletal development retardation in mice. AB - Maternal infection is a cause of adverse developmental outcomes. Lipopolysaccharide (LPS)-induced embryonic resorption, intra-uterine fetal death (IUFD) and preterm labor have been well characterized. In the present study, we investigated the effects of maternal LPS exposure on intra-uterine fetal growth and skeletal development. All pregnant mice except controls received an intraperitoneal injection of LPS (75 microg/kg) on gestational days (GD) 15-17. The number of live fetuses, dead fetuses and resorption sites was counted on GD 18. Live fetuses in each litter were weighed. Crown-rump and tail lengths were examined and skeletal development was evaluated. As expected, perinatal LPS exposure resulted in 63.2% fetal death. LPS significantly lowered fetal weight, reduced crown-rump and tail lengths, and retarded skeletal ossification in caudal vertebrae, anterior and posterior phalanges, and supraoccipital bone. Additional experiment showed that a single dose of LPS (75 microg/kg, i.p.) on GD 15 increased the expression of TNF-alpha mRNA in maternal liver and placenta and TNF alpha concentration in maternal serum and amniotic fluid. Furthermore, pentoxifylline, an inhibitor of TNF-alpha synthesis, significantly inhibited TNF alpha production, reduced fetal mortality, and reversed LPS-induced fetal intra uterine growth restriction and skeletal development retardation. Taken together, these results suggest that TNF-alpha is, at least in part, involved in LPS induced intra-uterine fetal death, intra-uterine growth restriction and skeletal development retardation. PMID- 16263229 TI - Preparation and evaluation of reverse-phase evaporation and multilamellar niosomes as ophthalmic carriers of acetazolamide. AB - Niosomes have been reported as a possible approach to improve the low corneal penetration and bioavailability characteristics shown by conventional ophthalmic vehicles. Niosomes formed from Span 40 or Span 60 and cholesterol in the molar ratios of 7:4, 7:6 and 7:7 were prepared using reverse-phase evaporation and thin film hydration methods. The prepared systems were characterized for entrapment efficiency, size, shape and in vitro drug release. Stability studies were carried out to investigate the leaching of drug from niosomes during storage. The intraocular pressure (IOP) lowering activity of acetazolamide niosomal formulations in rabbits was measured using ShiOtz tonometer. Histological examination for the corneal tissues of rabbits receiving niosomal formulations was carried out for assessment of the ocular irritancy of niosomes. The results showed that the type of surfactant, cholesterol content and the method of preparation altered the entrapment efficiency and drug release rate from niosomes. Higher entrapment efficiency was obtained with multilamellar niosomes prepared from Span 60 and cholesterol in a 7:6 molar ratio. Niosomal formulations have shown a fairly high retention of acetazolamide inside the vesicles (approximately 75%) at a refrigerated temperature up to a period of 3 months. Each of the tested acetazolamide niosomes prepared by either method produced a significant decrease in IOP compared to the solution of free drug and plain niosomes. Multilamellar acetazolamide niosomes formulated with Span 60 and cholesterol in a 7:4 molar ratio were found to be the most effective and showed prolonged decrease in IOP. Histological examination of corneal tissues after instillation of niosomal formulation for 40 days showed slight irritation in the substantia propria of the eye which is reversible and no major changes in tissues were observed. PMID- 16263230 TI - On the prediction of sweep rate and directional selectivity for FM sounds from two-tone interactions in the inferior colliculus. AB - Two-tone stimuli have traditionally been used to reveal regions of inhibition in auditory spectral receptive fields, particularly for neurons with low spontaneous rates. These techniques reveal how different frequencies excite or suppress the response to an excitatory frequency of a cell, but have often been assessed at a fixed masker-probe time interval. We used a variation of this methodology to determine whether two-tone spectrotemporal interactions can account for rate dependent directional selectivity for frequency modulations (FM) in the mustached bat inferior colliculus (IC). First, we quantified the response to upward and downward sweeping, linear, fixed-bandwidth FM tones centered at a unit's characteristic frequency (CF) at 6 sweep durations ranging from 2 to 64 ms. Then, to examine how responses to instantaneous frequencies contained within the sweeps might interact in time, we varied the frequency and relative onset of a brief (4 ms) "conditioner" tone paired with a fixed 4-ms CF probe tone. We constructed "conditioned response areas" (CRA) depicting regions of suppression and facilitation of the probe tone caused by the conditioning tone. We classified the CRAs as predominantly excitatory (40.9%), inhibitory (22.7%), or mixed (36.4%). To generate FM response predictions, the CRAs were multiplied with spectrograms of the same sweeps used to assess response to FM. The predictions of FM rate and directionality were accurate by our criteria in approximately 20% of units. Conversely, the CRAs from the remaining units failed to predict FM responses as accurately, suggesting that most IC units respond differently to FM sweeps than they do to tone-pairs matched to the instantaneous frequencies contained in those sweeps. The implications of these results for models of FM directionality are discussed. PMID- 16263231 TI - A real-time combined polymerase chain reaction assay for the rapid detection and differentiation of Anaplasma phagocytophilum, Ehrlichia chaffeensis, and Ehrlichia ewingii. AB - A rapid real-time polymerase chain reaction (PCR) assay capable of the simultaneous detection and differentiation of Anaplasma phagocytophilum, Ehrlichia chaffeensis, and Ehrlichia ewingii was developed using the LightCyclertrade mark instrument (Roche Applied Sciences, Indianapolis, IN). The assay targets the operon groEL of the heat shock protein. Base pair mismatches in amplified DNA in regions of detection probe hybridization allowed organism differentiation by melting curve analysis. The analytical sensitivity was at least 10 copies per reaction. DNA extracts from 59 specimens previously confirmed positive for A. phagocytophilum (n = 37), E. chaffeensis (n = 19), or E. ewingii (n = 3) were used to evaluate the assay. All of the specimens positive for 1 of the 3 organisms by conventional PCR were likewise positive by the LightCycler method. Sensitivity and specificity were at least 100% compared with conventional PCR. This assay provides a rapid method for the detection and differentiation of the causative agents of human ehrlichiosis in the United States. PMID- 16263232 TI - Evaluation of Czapeck agar and Sabouraud dextrose agar for the culture of airborne Aspergillus conidia. AB - We compared the efficiency of Czapeck agar and Sabouraud dextrose agar irradiated to detect Aspergillus conidia in air. Both media were equally suitable. Czapeck agar should be used in institutions interested only in Aspergillus surveys, and Sabouraud agar should be used in those institutions interested in all sorts of filamentous fungi. PMID- 16263233 TI - Antimicrobial activity of daptomycin tested against clinical strains of indicated species isolated in North American medical centers (2003). AB - Daptomycin is a cyclic lipopeptide approved for use by the US Food and Drug Administration (FDA) for the treatment of complicated skin and skin structure infections caused by Staphylococcus aureus, groups A and B beta-hemolytic streptococci, and vancomycin-susceptible Enterococcus faecalis. We evaluated the daptomycin spectrum against these pathogens by testing 2759 clinical strains consecutively collected in more than 30 hospitals located across the United States and Canada. The isolates were susceptibility tested against daptomycin and many comparators by the reference broth microdilution method. Daptomycin was very active against indicated species with the highest MIC results being 1, 2, and 0.5 microg/mL for S. aureus, E. faecalis, and beta-hemolytic streptococci, respectively. All isolates tested were considered susceptible to daptomycin, according to Clinical and Laboratory Standards Institute and FDA breakpoints, and daptomycin was the most potent (lowest MIC90) among selected antimicrobials commonly used to treat Gram-positive infections. Resistance to oxacillin or vancomycin did not influence daptomycin activity against S. aureus or E. faecalis. PMID- 16263234 TI - Modeling conversion and transport phenomena in solid-state fermentation: a review and perspectives. AB - Solid-state fermentation (SSF) is accompanied inevitably by development of concentration and temperature gradients within the substrate particles and microbial biofilms. These gradients are needed for driving the transport of substrates and products. In addition, concentration gradients have been suggested to be crucial for obtaining the characteristics that define the products of SSF; nevertheless, gradients are also known to result in reduced productivity and unwanted side reactions. Solid-state fermentations are generally batch processes and this further complicates their understanding as conditions change with time. Mathematical models are therefore needed for improving the understanding of SSF processes and allowing their manipulation to achieve the desired outcomes. Existing models of SSF processes describe coupled substrate conversion and diffusion and the consequent microbial growth. Existing models disregard many of the significant phenomena that are known to influence SSF. As a result, available models cannot explain the generation of the numerous products that form during any SSF process and the outcome of the process in terms of the characteristics of the final product. This review critically evaluates the proposed models and their experimental validation. In addition, important issues that need to be resolved for improved modeling of SSF are discussed. PMID- 16263235 TI - [Extrapontine myelinolysis after correction of hyponatremia]. PMID- 16263236 TI - Synthesis and in-vitro cytotoxicity evaluation of gatifloxacin Mannich bases. AB - Mannich bases of gatifloxacin were synthesized by reacting them with formaldehyde and several isatin derivatives. Their chemical structures have been confirmed by means of their IR, 1H-NMR data and by elemental analysis. The compounds were tested in-vitro against a panel of 58 human tumour cell lines derived from nine neoplastic diseases. Among them compound 1-cyclopropyl-6-fluoro-8-methoxy-1,4 dihydro-4-oxo-7[[N4-(3'-sulphadoximino)-1'-(5-bromoisatinyl) methyl]-3-methyl N1 piperazinyl]-3-quinoline carboxylic acid (6) emerged as a potent anticancer agent being more active than standard DNA topoisomerase II inhibitor, etoposide against 30 cancer cell lines. PMID- 16263237 TI - Polypodium leucotomos extract: antioxidant activity and disposition. AB - The extract of the fern Polypodium leucotomos (PL, Fernblock) is an oral photoprotectant with strong antioxidative properties. Recent studies to determine its chemical composition have shown 4-hydroxycinnamic acid (p-coumaric), 3 methoxy-4-hydroxycinnamic acid (ferulic), 3,4-dihydroxycinnamic acid (caffeic), 3 methoxy-4-hydroxybenzoic acid (vanillic) and 3-caffeoilquinic acid (chlorogenic) to be among its major phenolic components. No conclusive data are available, however, on the H2O2-scavenging capacity of these compounds, or on their absorption and metabolism following their oral intake. In the present work, their antioxidative capacity was assessed by the luminol/H2O2 assay, their absorption studied using Caco-2 cells to resemble the intestinal barrier, and their metabolism investigated using cultured primary rat hepatocytes. The antioxidant capacity of PL components increased in a concentration-dependent manner, with ferulic and caffeic acids the most powerful antioxidants. The apparent permeability results correspond to a human post-oral administration absorption of 70-100% for all tested substances. Coumaric, ferulic and vanillic acids were metabolized by CYP450-dependent mono-oxygenases and partially conjugated to glucuronic acid and sulfate. These phenolic compounds may contribute to the health benefits afforded by this oral photoprotectant. PMID- 16263238 TI - The approximately 30-million-year-old ERVPb1 envelope gene is evolutionarily conserved among hominoids and Old World monkeys. AB - Most human endogenous retroviruses (HERVs) are ancient and their genes are rendered nonfunctional by debilitating mutations. One exception is a recently discovered envelope gene located on chromosome 14. This envelope protein was also recently shown to be expressed in various human tissues and to mediate cell-cell fusion ex vivo. In this study, we demonstrate that this locus (designated ERVPb1) is preserved in Old World monkeys and that the reading frame is maintained. This is congruent with the entry of the HERV-P(b) group between 27 and 36 million years ago as suggested by long terminal repeat divergence. Although the coding capacity is generally lost in the HERV-IP supergroup, the analysis of nucleotide substitutions, lack of stop codons, and single-nucleotide polymorephisms strongly indicates a selective advantage of the ERVPb1 envelope genes during primate evolution. The purifying selection and tissue-specific expression of the human ERVPb1 envelope gene provide strong evidence of a beneficial role for the host. PMID- 16263239 TI - Comparison of verbal preference assessments in the presence and absence of the actual stimuli. AB - Stimulus preference assessments for individuals with developmental disabilities typically involve offering choices among stimuli and providing immediate access to the chosen stimuli. Several researchers have explored the utility of presenting choices verbally, thereby obviating the need to present the choices in tangible form and deliver access to those choices immediately. However, studies that have compared verbal selection to selection among tangible stimuli have nonetheless delivered the chosen stimulus following selections, in essence, manipulating the antecedent but not the consequence. It therefore remains unclear whether preference assessments that do and do not include the actual stimuli yield comparable results. The current study compared preference assessment results for three participants in which either (a) the stimuli were presented, selections were made verbally, and selection resulted in no differential consequence, or (b) the stimuli were presented both verbally and in tangible form, and selection produced access to the stimulus. Reinforcer assessments were then conducted to test contradictory predictions of reinforcer efficacy made by the two methods. Comparisons between the two assessments yielded only modest rank order correlations (M=0.24; range, -0.17 to 0.57) that varied widely across participants. Results of the reinforcer assessments suggested that the verbal plus-tangible stimulus preference assessment more accurately predicted reinforcer strength. PMID- 16263241 TI - Robust self-localisation and navigation based on hippocampal place cells. AB - A computational model of the hippocampal function in spatial learning is presented. A spatial representation is incrementally acquired during exploration. Visual and self-motion information is fed into a network of rate-coded neurons. A consistent and stable place code emerges by unsupervised Hebbian learning between place- and head direction cells. Based on this representation, goal-oriented navigation is learnt by applying a reward-based learning mechanism between the hippocampus and nucleus accumbens. The model, validated on a real and simulated robot, successfully localises itself by recalibrating its path integrator using visual input. A navigation map is learnt after about 20 trials, comparable to rats in the water maze. In contrast to previous works, this system processes realistic visual input. No compass is needed for localisation and the reward based learning mechanism extends discrete navigation models to continuous space. The model reproduces experimental findings and suggests several neurophysiological and behavioural predictions in the rat. PMID- 16263240 TI - Hippocampal mechanisms for the context-dependent retrieval of episodes. AB - Behaviors ranging from delivering newspapers to waiting tables depend on remembering previous episodes to avoid incorrect repetition. Physiologically, this requires mechanisms for long-term storage and selective retrieval of episodes based on the time of occurrence, despite variable intervals and similarity of events in a familiar environment. Here, this process has been modeled based on the physiological properties of the hippocampal formation, including mechanisms for sustained activity in entorhinal cortex and theta rhythm oscillations in hippocampal subregions. The model simulates the context-sensitive firing properties of hippocampal neurons including trial-specific firing during spatial alternation and trial by trial changes in theta phase precession on a linear track. This activity is used to guide behavior, and lesions of the hippocampal network impair memory-guided behavior. The model links data at the cellular level to behavior at the systems level, describing a physiologically plausible mechanism for the brain to recall a given episode which occurred at a specific place and time. PMID- 16263243 TI - Immunopathogenic role of TH1 cells in autoimmune diabetes: evidence from a T1 and T2 doubly transgenic non-obese diabetic mouse model. AB - To improve the feasibility of in vivo monitoring of autoreactive T cells in the diabetogenic process, we generated T1 and T2 doubly transgenic non-obese diabetic (NOD) mice in which transgenic human CD90 (hCD90) is simultaneously expressed on IFN-gamma-producing cells or murine CD90.1 (mCD90.1) is expressed on IL-4 producing cells. These transgenic NOD mice develop diabetes with the same kinetics and incidence as wild type NOD mice, permitting the physiological characterization of CD4(+)hCD90(+) cells, which represent T(H)1 cells in lymphoid organs and at the site of insulitis. CD4(+)hCD90(+) cells had a higher capacity to secret IFN-gamma than CD4(+)hCD90(-) cells in an autoantigen-specific manner. Transgenic mice treated with GAD65 plasmid were protected from autoimmune diabetes, and had a lower number of CD4(+)hCD90(+) cells, confirming the pathogenic role of CD4(+)hCD90(+) cells in autoimmune diabetes. To further investigate the effect of IL-12 on the development of T(H)1 cells in autoimmune diabetes, we crossed these doubly transgenic mice to IL-12p35-deficient NOD mice. Despite severe disturbance of diabetes in p35(-/-) mice, the frequency of T(H)1 cells in these mice was slightly lower than in wild type mice. These data support the pathological role of IL-12 in autoimmune diabetes and suggest the existence an IL-12-independent pathway of T(H)1 development. PMID- 16263242 TI - GAD65- and proinsulin-specific CD4+ T-cells detected by MHC class II tetramers in peripheral blood of type 1 diabetes patients and at-risk subjects. AB - In type 1 diabetes the major loss of insulin producing beta-cells is caused by autoreactive T-cells specific for antigens expressed by the pancreatic islets. In this study we have analyzed the prevalence of glutamate decarboxylase 65 (GAD65)- and proinsulin-specific CD4(+) T-cells in type 1 diabetes patients, at-risk subjects and in HLA-matched control children. Peripheral blood mononuclear cells were cultured in the presence of two different GAD65 peptides (555-567, 557I and 274-286) or with a proinsulin (B24-C36) peptide for 10-11days. The autoreactive T cells were detected using antigen specific-MHC class II tetramers by flow cytometry. Our results show that 11 of 18 (61%) type 1 diabetes patients and 7 of the 20 (35%) at-risk subjects were positive for one of the three GAD65 or proinsulin-containing tetramers, whereas only 2 of 21 (9.5%) controls had tetramer binding cells (p = 0.0007 type 1 diabetes vs. controls and p = 0.0488 at risk subjects vs. controls, Chi-square test). Type 1 diabetes patients responded to all three peptides. At-risk subjects recognized also the GAD65 555-567 557I peptide, while none of the controls responded to it. In conclusion, type 1 diabetes patients and at-risk subjects have a significantly higher prevalence of GAD65- and proinsulin-specific CD4(+) T-cells than the control subjects. PMID- 16263244 TI - Immunopathogenesis of psoriasis: recent insights on the role of adaptive and innate immunity. AB - Psoriasis is a frequent chronic inflammatory disorder involving mostly skin and joints. Its characteristic features in the skin consist of inflammatory changes in both dermis and epidermis, with abnormal keratinocytic differentiation and proliferation. In recent years, an important set of knowledge has been provided by works addressing the immunopathogenic mechanisms of the disease. Indeed, recent advances in the knowledge of mechanisms linking innate and adaptive immunity have led to reconsideration of the roles of key players in the pathogenesis of the disease. This review will focus on results from studies performed in vitro and in vivo in patients with psoriasis, and on lessons from recently designed animal models which are considered as the most relevant with respect to the human disease. Even more important, these insights provide a rational basis for the design of new therapeutic strategies aiming at the deletion or the down regulation of activated T cells, or at the suppression of pathogenic cytokines such as TNF-alpha. Some of these new biotherapeutic tools have been successfully used in vivo in clinical trials, providing a confirmation of such concepts. PMID- 16263245 TI - The genetics of type 1 diabetes: lessons learned and future challenges. AB - It has been more than 30 years since the first evidence was published suggesting the involvement of a specific chromosomal region, HLA, in modulating the risk for type 1 diabetes (T1D). In the intervening years, what have we learned regarding the identities of specific loci that modulate T1D risk, and what lessons have these studies provided that might be helpful in finding and characterizing additional susceptibility loci both for T1D and other autoimmune disorders? In the following review, we briefly address these issues. PMID- 16263246 TI - Treatment of a first episode of psychotic illness with quetiapine: an analysis of 2 year outcomes. AB - BACKGROUND: The first episode of a psychotic disorder provides a unique opportunity to initiate optimal treatment but when a new medication becomes available, little data exist to guide the appropriate use in this population. OBJECTIVES: The objectives were to determine the optimal doses and titration of quetiapine for this group and to measure outcomes (including symptom response, social functioning, mood alterations, motor symptoms, metabolic parameters and cognitive functioning) over 2 years of treatment with quetiapine. DESIGN: Thirty nine subjects with a first episode of psychosis referred to the Nova Scotia Early Psychosis Program in Halifax, Canada, were invited to participate in this study. Standardized clinical, laboratory, and neuropsychological assessments were performed at baseline and following treatment with quetiapine at intervals out to 2 years. RESULTS: Quetiapine was effective in treating the psychotic and mood symptoms while not causing extra-pyramidal signs or symptoms (EPSS). Pre-existing motor dysfunction improved. No anticholinergic medications were required. Several domains of cognitive function also improved (sustained attention, the number of perseverative errors, visuomotor speed and sequencing, verbal fluency and verbal memory). Weight gain was observed along with increases in cholesterol levels but there was no glucose dysregulation. CONCLUSIONS: The results of this two year, naturalistic study of people with a first episode of psychosis indicated that quetiapine was well tolerated and effective for this population. Significant improvements in cognitive functioning also provided evidence for potential longer term benefits of early and optimal treatment with this agent. However, monitoring metabolic parameters, as recommended for other atypicals, is likely prudent. PMID- 16263247 TI - Prevalence of diabetes mellitus in chronic schizophrenic inpatients in relation to long-term antipsychotic treatment. AB - BACKGROUND: Many reports indicate that the incidence and prevalence of diabetes mellitus is increased in schizophrenic patients and related to antipsychotic treatment. In an exploratory cross-sectional study we assessed the prevalence of type 2 diabetes mellitus in 266 chronic schizophrenic and schizoaffective inpatients and investigated whether the duration of antipsychotic treatment was related to the development of diabetes mellitus. METHOD: We measured the non fasting plasma glucose level in 266 inpatients with DSM IV diagnosis of schizophrenia or schizoaffective disorder in 5 different long-stay wards in the Netherlands. Measured variables were: age, sex, ethnicity, BMI, current antipsychotic treatment, duration of illness and duration of antipsychotic treatment. RESULTS: The overall prevalence of type 2 diabetes mellitus was 9%, which is significantly higher than the prevalence of 4.9% in the general population (OR 1.89, CI 1.14-3.13; p<0.014). The prevalence was increased in two age cohorts: 30-39 years (3.8% vs. 0.3%, OR=13.29, CI=2.17-81.36, p=<0.005) and 40-49 years (9.3% vs. 1.5%, OR=6.74, CI=2.77-16.38, p=0.000). No new cases of diabetes mellitus were detected during the course of the study. The increased prevalence was found to be related to overweight and obesity. The time of exposure to antipsychotic treatment was not significantly correlated with the prevalence of diabetes mellitus when adjusted for age (F=0.804, df=1, p=0.371, respectively, F=0.194, df=1, p=0.660). Both typical and atypical antipsychotics contributed equally to the prevalence of diabetes mellitus. CONCLUSION: No significant relation between long-term antipsychotic treatment and prevalence of diabetes mellitus was found. The high prevalence of diabetes mellitus in schizophrenic patients warrants screening of these patients already at young age for glucose disturbance. PMID- 16263248 TI - Compensatory recruitment after sleep deprivation and the relationship with performance. AB - This study examined the effects of total sleep deprivation (TSD) on cerebral responses to a verbal learning task with two levels of word difficulty. A total of 32 subjects were studied with functional magnetic resonance imaging (FMRI) after normal sleep and following 36 h of TSD. Cerebral responses to EASY words were identical on both nights, but several brain regions showed increased activation to HARD words following TSD compared with following a normal night of sleep (NORM). These regions included bilateral inferior frontal gyrus, bilateral dorsolateral prefrontal cortex, and bilateral inferior parietal lobe. Better free recall performance on the HARD words after TSD was related to increased cerebral responses within the left inferior and superior parietal lobes and left inferior frontal gyrus. Recall was negatively related to activation within the right inferior frontal gyrus. Overall, the findings support the predictions of the compensatory recruitment hypothesis that task demands influence both the likelihood and location of increased cerebral activation during task performance following TSD, and refine that hypothesis by identifying a specific task demand that plays a role. The performance relationships suggest increased activation may be both beneficial (compensatory) and interfere with task performance, depending on the brain regions involved. PMID- 16263249 TI - Measurement of the local 1H spin-diffusion coefficient in polymers. AB - Proton spin diffusion is widely used to determine domain sizes in heterogeneous organic solids. For an accurate analysis, spin diffusion coefficients are required. However, in most cases they are not directly measured, but instead derived from model systems. The effects of magic-angle spinning (MAS), mobility, or spin-lock fields on spin-diffusion coefficients have also been difficult to quantify. In this work, direct measurement of local (1)H spin-diffusion coefficients in any rigid polymer is achieved in experiments with heteronuclear dephasing of the (1)H magnetization, a mixing time for (1)H spin diffusion, and (13)C detection after cross-polarization. In the presence of (1)H homonuclear decoupling and (13)C 180 degrees-pulse recoupling, each (13)C spin dephases a significant number (3-20) of protons, depending on the dephasing time. For (13)C and other sufficiently dilute heteronuclei, the dephasing of the protons is described by simple spin-pair REDOR curves. As a result, every (13)C nucleus will "burn" a spherical hole of known diameter and profile into the proton magnetization distribution. (1)H spin diffusion into the hole during the mixing time can be monitored and simulated accurately for every resolved (13)C site, with the spin-diffusion coefficient as the only significant unknown parameter. By varying the dephasing time, holes with diameters of 0.4-0.8 nm can be burned into the proton magnetization profile and thus the dependence of the local spin diffusion coefficients on the proton density or partial mobility can be explored. The effects of transverse or magic-angle spin-lock fields on spin diffusion can be quantified conveniently by this method. Analytical and numerical fits yield short-range spin-diffusion coefficients of 0.2-0.5 nm(2)/ms on the 0.5-nm scale, which is smaller than the value of 0.8 nm(2)/ms for organic solids previously measured on the 10-nm scale. PMID- 16263250 TI - NMR spectroscopy of hydrogen adsorption on single-walled carbon nanotubes after exposure to high pressure. AB - Hydrogen storage properties of single-walled carbon nanotubes (CNTs) after exposure to a pressure of 14.3 MPa are studied by (1)H nuclear magnetic resonance spectroscopy. The nanotubes were carefully pre-characterized using inductively coupled plasma mass spectrometry (ICP-MS), transmission electron microscopy (TEM), and Raman spectroscopy. We have shown previously that at ambient temperature in the pressure range from 0 to 1.5 MPa, hydrogen adsorption is fast and reversible and must be described as physisorption. However, exposure to a much higher pressure (14.3 MPa) of hydrogen leads to slower desorption kinetics where longer exposure causes greater hydrogen uptake. Our data suggest that interstitial sites and the tube interior may be identified as these strong adsorption sites. PMID- 16263251 TI - Assessment of the effects of the cyclooxygenase-2 inhibitor rofecoxib on visuospatial learning and hippocampal cell death following kainate-induced seizures in the rat. AB - Kainate-induced seizures result in hippocampal neurodegeneration and spatial learning deficits in rodents. Previous studies show that rofecoxib, a selective cyclooxygenase-2 inhibitor, protects against kainate-induced hippocampal cell death 3 days after seizures. Our aim was to determine whether rofecoxib attenuates visuospatial learning deficits and late neuronal death after kainate induced seizures. Seizures were induced in Sprague-Dawley rats with kainic acid (10 mg/kg, i.p.). Eight hours later, animals received rofecoxib (10 mg/kg; n = 15) or vehicle (dimethylsulfoxide, n = 11). Animals were then treated daily for additional 2 or 9 days. Visuospatial learning was assessed in the Morris water maze (MWM) on days 5-9 after seizures. Seizure animals learned the MWM task significantly slower than non-seizure controls, but seizure animals showed higher swim speed (P < 0.05). Seizure animals receiving rofecoxib for 2 days showed no significant improvement in acquisition of the task compared to the vehicle group, even though mean latencies in the rofecoxib group were shorter from the third trial day onwards. This tendency was lost when rofecoxib was given for 9 days. TdT-mediated dUTP nick end labelling showed cell death in limbic structures 9 days after seizures. The time course of kainate-induced hippocampal cell death might be delayed by rofecoxib treatment, as the attenuation of cell death observed 3 days after seizures was no longer present after 9 days. We conclude that even though increasing evidence points to an injurious role of cyclooxygenase-2 products in acute brain injury processes, rofecoxib treatment failed to attenuate seizure-induced visuospatial learning deficits and the late phase of hippocampal neurodegeneration. PMID- 16263252 TI - Interactions between the dopamine agonist, bromocriptine and the efflux protein, P-glycoprotein at the blood-brain barrier in the mouse. AB - Bromocriptin (BCT) is a dopaminergic receptor agonist, poorly transported through the blood-brain barrier (BBB) and responsible for central side effects. Interactions between BCT and the efflux protein, P-glycoprotein (Pgp), have been described in vitro but nothing is known in vivo nor at the BBB level. At the BBB, in vivo, we investigated BCT as (i) a Pgp substrate by comparing the brain uptake in CF1 mdr1a(-/-) and mdr1a(+/+) mice with or without inhibitors of Pgp (valspodar, elacridar); (ii) a Pgp inducer by looking at the effect of repeated doses of BCT on cerebral uptake of digoxin and comparing it to the effect of dexamethasone and rifampicin; (iii) a Pgp inhibitor by determining the effect of a single dose of BCT on cerebral uptake of digoxin and comparing it to the effect of valspodar. CF1 mdr1a(-/-) mice showed much higher brain uptake of BCT than CF1 mdr1a(+/+) mice and brain uptake of BCT was higher in CF1 mdr1a(+/+) mice pre treated with valspodar or elacridar indicating that BCT is a Pgp substrate at the BBB level. Brain uptake of digoxin was not modified in CF1 mdr1a(+/+) mice pre treated with a single dose or repeated doses of BCT, indicating that BCT is neither a Pgp inductor nor a Pgp inhibitor at the BBB in the chosen experimental setting. In vivo, at the mouse BBB level and in our experimental conditions, bromocriptin is a Pgp substrate but is not a Pgp modulator. PMID- 16263253 TI - Secreted protein acidic and rich in cysteine (SPARC/osteonectin/BM-40) binds to fibrinogen fragments D and E, but not to native fibrinogen. AB - Secreted protein acidic and rich in cysteine (SPARC/osteonectin/BM-40) is a matricellular protein that functions in wound healing. Fibrinogen is a plasma protein involved in many aspects of wound healing, such as inflammation, fibrosis and thrombosis. In this study, the binding of SPARC to both native and plasmin cleaved fibrinogen under physiological conditions was examined by the use of a surface plasmon resonance (SPR) biosensor. We show that SPARC binds to plasmin cleaved fibrinogen, but not to native fibrinogen. SPARC binds to both fibrinogen fragments D and E fg D and fg E with similar dissociation constants (8.67 x 10( 8) M for Fg D and 1.61 x 10(-7) M for Fg E). Results from endothelial cell proliferation assays show that the binding of SPARC to Fg E suppressed the inhibition of proliferation by SPARC, whereas the binding of SPARC to Fg D did not influence the activity of SPARC on the cell cycle. The interaction of SPARC with fibrinogen fragments D and E, which are produced as a result of proteolytic activation of fibrinolysis, reveals potential storage sites in provisional extracellular matrix for SPARC during the wound healing process and indicates a regulatory role of SPARC in fibrinolysis and angiogenesis. PMID- 16263254 TI - Tachyphylaxis to beta2-agonists in Spanish asthmatic patients could be modulated by beta2-adrenoceptor gene polymorphisms. AB - BACKGROUND: The study of determinants of asthma is a subject of much interest currently, especially the pharmacogenetic aspects of asthma management. Genetic polymorphisms affecting amino-acids at positions 16 and 27 within beta(2) adrenoceptor (beta(2)AR) gene have been implicated in the asthma phenotypes and influence on the variability observed in response to use of bronchodilator agents used in the treatment of asthma. Whether these polymorphisms alter the bronchoprotection response to beta(2)-agonist treatment in Spanish asthmatic population is unknown. The aim of this study was to investigate whether genetic polymorphisms within beta(2)AR gene modulate the clinical outcomes of the individual response to beta(2)-agonist therapy and the development of desensitization in Spanish asthmatic patients. METHODS: In a prospective, case control study were included 80 asthmatic patients. Based on the standard criteria, patients were classified into two groups: patients with tachyphylaxis and good responders to beta(2)-agonist therapy. DNA samples were genotyped for the Arg(16)Gly and Glu(27)Gln alleles within the beta(2)AR gene as well as in 64 control samples from blood donors. RESULTS: Arg(16) allele was slightly more frequent within the group with tachyphylaxis (P=0.039), whereas Gly(16) allele carriers were overrepresented within the group of good responders (59.7%, P=0.028). On the other hand, the allele frequency of Gln(27) and the proportion of Gln(27) carriers was higher within the group with tachyphylaxis (P=0.010 and 0.049, respectively) and Glu(27) allele carriers were overrepresented within the group of good responders (P=0.026). The Arg(16) and Gln(27) alleles were in strong linkage disequilibrium across this locus, resulting in the occurrence of disease haplotype. CONCLUSIONS: The predisposition to develop tachyphylaxis in our population seems to be linked to the Arg(16) and Gln(27) alleles and to the Arg(16)/Gln(27) risk haplotype (positive association between the presence of the Arg(16) and Gln(27) alleles and tachyphylaxis). The Arg(16) allele is perhaps overrepresented due to the strong linkage disequilibrium between both polymorphisms. The presence of the Glu(27) allele seems to be a protective factor against tachyphylaxis in this cohort study. PMID- 16263255 TI - Beneficial influence of dietary curcumin, capsaicin and garlic on erythrocyte integrity in high-fat fed rats. AB - In rats rendered hyperlipidemic by maintaining them on a high-fat diet (30%) for 8 weeks, inclusion of spice principles [curcumin (0.2%) or capsaicin (0.015%)] or garlic (2.0%) in the diet produced significant hypotriglyceridemic effect. Plasma cholesterol remained unaffected in high-fat treatment. Hepatic triglyceride content was significantly higher in high-fat fed rats, and this increase was effectively countered by inclusion of the hypolipidemic spice agents -- curcumin, capsaicin or garlic in the diet. The lipid profile of erythrocyte membranes of hyperlipidemic rats was similar to basal controls. An examination of the osmotic fragility of erythrocytes in various groups indicated that the red blood cells of hyperlipidemic rats display a slight resistance to osmotic lysis. Inclusion of spice principles [curcumin (0.2%) or capsaicin (0.015%)] or garlic (2.0%) in the diet, which produced the hypotriglyceridemic effect, appeared to beneficially correct this altered osmotic fragility of erythrocytes. Activities of ouabain sensitive Na(+),K(+)-ATPase as well as acetylcholinesterase of erythrocyte membranes in high-fat fed rats remained unaltered. Activity of Ca(2+),Mg(2+) ATPase in erythrocyte membrane was significantly decreased in high-fat fed animals, whereas dietary spice principles and garlic countered this reduction in enzyme activity. In the absence of any change in the cholesterol/phospholipid molar ratio in the erythrocyte membrane, a decreased activity of membrane-bound Ca(2+),Mg(2+)-ATPase could have probably contributed to the accumulation of intracellular calcium leading to the diminished deformability of the erythrocytes in high-fat fed rats. PMID- 16263256 TI - Bioluminescence technology for imaging cell proliferation. AB - As continuous cell proliferation caused by genetic alterations leads to cancer, monitoring abnormal cell proliferation in sporadic tumor models is important in the context of tumor generation, development and response to therapy. Bioluminescence imaging technology, which visualizes the conversion of chemical energy into visible light by luciferase enzymes, is an established method to measure cell numbers in grafted tumors in vivo, but has not been used to monitor cell proliferation per se. To measure cell proliferation noninvasively, transgenic mice have been developed that express the luciferase gene under the control of the E2F1 promoter. When these reporter mice are crossed with genetically defined mouse models of human cancer, the proliferative activity of the tumor cells can be monitored with proportional light production. These technologies support more detailed preclinical trials and could enable other biological pathways to be monitored in living cells. PMID- 16263257 TI - Innovation and greater probability of success in drug discovery and development - from target to biomarkers. PMID- 16263258 TI - Non-biodegradable biopolymers from renewable resources: perspectives and impacts. AB - In recent years the biotechnological production of bulk biopolymers has focused on the synthesis of biodegradable polymers to replace their non-biodegradable counterparts derived from fossil resources. Examples include polyhydroxyalkanoates and polylactic acid, which act as substitutes for polyolefins. By contrast, the biotechnological production of non-biodegradable polymers from renewable resources has so far been scarcely considered, probably because this idea contradicts the paradigm that all natural compounds are biodegradable. Polythioesters, which were recently described as new biopolymers, do not follow this paradigm because although they are produced by bacteria, they are persistent to microbial degradation. Mankind has a need for both non biodegradable and biodegradable polymers and methods to produce them from renewable resources will be of great value. PMID- 16263259 TI - Nanotechnology for drug and gene therapy: the importance of understanding molecular mechanisms of delivery. AB - Nanotechnology, although not a new concept, has gained significant momentum in recent years. This stems partly from the realization that nanosystems have significantly different biological properties from large-sized systems (e.g. implants or microparticles) that could be used effectively to overcome problems in drug and gene therapy. In drug therapy, we face the problems of inefficacy or nonspecific effects; hence, nanosystems are being developed for targeted drug therapy. In gene therapy using non-viral systems, the main issues are relatively transient gene expression and lower efficiency than viral vectors. Research efforts have focused on understanding the barriers in gene delivery so that non viral systems can be developed that are as effective as viral systems in gene transfection. Understanding the molecular mechanisms that underlie the interactions of nanosystems with the cell, their uptake properties and retention will be crucial for the successful development of these systems. PMID- 16263260 TI - The avian 'prefrontal cortex' and cognition. AB - Both mammals and birds can flexibly organize their behavior over time. In mammals, the mental operations generating this ability are called executive functions and are associated with the prefrontal cortex. The corresponding structure in birds is the nidopallium caudolaterale. Anatomical, neurochemical, electrophysiological and behavioral studies show these structures to be highly similar. The avian forebrain displays no lamination that corresponds to the mammalian neocortex, hence lamination does not seem to be a requirement for higher cognitive functions. Because all other aspects of the neural architecture of the mammalian and the avian prefrontal areas are extremely comparable, the freedom to create different neural architectures that generate prefrontal functions seems to be very limited. PMID- 16263261 TI - Place cells, spatial maps and the population code for memory. AB - The study of population dynamics in hippocampal place cells has emerged as one of the most powerful tools for understanding the encoding, storage and retrieval of declarative memory. Recent work has laid out the contours of an attractor-based hippocampal population code for memory in recurrent circuits of the hippocampus. The code is based on inputs from a topographically organized, path-integration dependent spatial map that lies upstream in the medial entorhinal cortex. The recurrent networks of the hippocampal formation enable these spatial inputs to be synthesized with nonspatial event-related information. PMID- 16263262 TI - Structuring the puzzle of prion propagation. AB - Of all the prion proteins identified to date, the agent responsible for transmissible spongiform encephalopathies is one of the least characterized. Nevertheless, recent advances in the prion field should lead to important progress in our knowledge of mammalian prions. First, the demonstration that PrP aggregates generated in vitro infect animals and cause neuronal death is a considerable breakthrough. Second, new structural data provide direct insight into the structure of the infectious agent. Third, the study of yeast prions unveiled what might be the structural basis for the strain phenomena in transmissible spongiform encephalopathies. PMID- 16263263 TI - Structure and activity of enzymes that remove histone modifications. AB - The post-translational modification of histones plays an important role in chromatin regulation, a process that insures the fidelity of gene expression and other DNA transactions. Equally important as the enzymes that generate these modifications are the enzymes that remove them. Recent studies have identified some of the enzymes that remove histone modifications and have characterized their activities. In addition, structural and biochemical studies of these enzymes have focused on the histone lysine deacetylases HDAC8 and sirtuins, and on the arginine and lysine demethylases PAD and BHC110/LSD1, respectively. These new findings may be used as a context to present new information that contributes to our understanding of chromatin regulation, and to pose remaining questions pertaining to the activities of these enzymes and the roles they play in chromatin regulation. PMID- 16263265 TI - The type III needle and the damage done. AB - Many Gram-negative pathogens translocate virulence proteins directly into host cells using a type III secretion system. This complex secretion machinery is composed of approximately 25 different proteins that assemble to span both bacterial membranes, and contact the host cell to form a direct channel between the bacterial and host cell cytoplasms. Assembly of the system and efficient secretion of virulence proteins through this apparatus require specific chaperones. Although the machinery is morphologically conserved among all bacteria, the secreted proteins vary widely and are responsible for the range of diseases caused by bacterial pathogens. Recent structures have given insights into important chaperone and effector proteins, as well as revealing the first atomic structures of portions of the secretion machinery itself. PMID- 16263264 TI - Structural biology of the thioester-dependent degradation and synthesis of fatty acids. AB - The fatty acid degradation and synthesis pathways consist of the same four chemical transformations. These transformations are facilitated by conjugating the fatty acid, via a thioester bond, to coenzyme A or acyl carrier protein in, respectively, the degradation and synthesis pathways. These pathways are compartmentalized in the peroxisomes, mitochondria and cytosol of eukaryotic cells. Current structural knowledge of the enzymes comprising these pathways shows that the approximately 130 entries in the RCSB Protein Data Bank can be grouped into seven superfamilies. Multifunctional enzymes are important in both pathways. PMID- 16263266 TI - Structural insights into SARS coronavirus proteins. AB - The SARS coronavirus was identified as the pathogen of a global outbreak of SARS (severe acute respiratory syndrome) in 2003. Its large RNA genome encodes four structural proteins, sixteen non-structural proteins and eight accessory proteins. The availability of structures of SARS coronavirus macromolecules has enabled the elucidation of their important functions, such as mediating the fusion of viral and host cellular membranes, and in replication and transcription. In particular, the spike protein fusion core and the main protease have been the most extensively studied, with the aim of designing anti-SARS therapeutics. Attention is now being focused on replicase proteins, which should enhance our understanding of the replication and transcription machinery. The structures and functions of most SARS proteins remain unknown, and further structural studies will be important for revealing their functions and for designing potential anti-SARS therapeutics. PMID- 16263267 TI - Regulation of apoptosis: uncovering the binding determinants. AB - Eukaryotic cells use complex networks of signal transduction proteins to make decisions about whether to differentiate, grow or die. In the case of apoptosis, which is responsible for the programmed death of unwanted or damaged cells in multicellular organisms, recent structural, biochemical and cell-based assays have enhanced our understanding of the mechanisms by which some of the key proteins regulate this process. These studies have highlighted a critical role for conformational change and the regulated formation of specific complexes that can either inhibit or stimulate apoptosis. In some cases, it is still not clear what distinguishes inhibitory from activating complexes, but the value of a structural understanding is highlighted by the success of recent structure-based drug discovery programs that have targeted these complexes. PMID- 16263268 TI - Recent structural insights into the expanding world of carbohydrate-active enzymes. AB - Enzymes that catalyse the synthesis and breakdown of glycosidic bonds account for 1-3% of the proteins encoded by the genomes of most organisms. At the current rate, over 12 000 glycosyltransferase and glycoside hydrolase open reading frames will appear during 2006. Recent advances in the study of the structure and mechanism of these carbohydrate-active enzymes reveal that glycoside hydrolases continue to display a wide variety of scaffolds, whereas nucleotide-sugar dependent glycosyltransferases tend to be grafted onto just two protein folds. The past two years have seen significant advances, including the discovery of a novel NAD+-dependent glycosidase mechanism, the dissection of the reaction coordinate of sialidases and a better understanding of the expanding roles of auxiliary carbohydrate-binding domains. PMID- 16263269 TI - Glutathione transferases: new functions. AB - Well known as detoxification enzymes, the glutathione transferases also function in prostaglandin and steroid hormone synthesis. New uses for the canonical glutathione transferase fold are becoming apparent; the bacterial stringent starvation protein SspA and the yeast prion protein Ure2p (both transcription factors) were found to adopt this fold, but their roles remain unclear. The intracellular chloride ion channel CLIC1 adopts the canonical glutathione transferase fold in its soluble form and appears to undergo radical structural modification as part of its membrane insertion process. The structures of rat and human mitochondrial glutathione transferases have been solved: they adopt a topology similar to that of bacterial disulfide bond isomerases, leading to the suggestion that they have evolved independently of the canonical enzymes. Recent structural studies of integral membrane glutathione S-transferases from microsomes have revealed common patterns of tertiary and quaternary structure. PMID- 16263270 TI - Enzymes in the biosynthesis of aromatic polyketide antibiotics. AB - Aromatic polyketides are secondary metabolites that afford some of the most common antibiotics and anti-cancer drugs currently in clinical use. Not least because of their medical importance, the biosynthesis of these compounds has attracted considerable interest during the past few years; important advances have been made in the structural and mechanistic characterisation of the enzymes involved. These studies are expected to have implications for the production of novel therapeutic agents by combinatorial biosynthesis. PMID- 16263271 TI - Orphan nuclear receptors adopted by crystallography. AB - Of the large nuclear hormone receptor superfamily of proteins, orphan nuclear receptors have remained a mystery owing to their lack of identified ligands and their constitutive nature. Now, structures of several ligand-binding domains of orphan receptors have provided some surprising insights that were not anticipated from molecular studies. Therefore, most orphan nuclear receptors have now been 'adopted' and their regulation has been shown to range from true ligand independence to highly promiscuous ligand-dependence. Former orphan receptors have been found to contain ligand-binding pockets that range in volume from vast (>1600A3) to non-existent and have been shown to generate surface AF2 motifs that range from being multifunctionally active to distinctly inactive. Insights from these new structures illustrate how powerful a structural biology approach can be when integrated with molecular and cellular physiology. PMID- 16263272 TI - Growth inhibition of estrogen receptor-positive and aromatase-positive human breast cancer cells in monolayer and spheroid cultures by letrozole, anastrozole, and tamoxifen. AB - Two third-generation aromatase inhibitors, letrozole and anastrozole, and the antiestrogen tamoxifen, were compared for growth-inhibiting activity in two estrogen receptor (ER)-positive aromatase-overexpressing human breast cancer cell lines, MCF-7aro and T-47Daro. Inhibition of hormone (1 nM testosterone) stimulated proliferation was evaluated in both monolayer cultures and in three dimensional spheroid cultures. Letrozole and anastrozole were also compared for effectiveness of aromatase inhibition, and relative affinity for aromatase, under both monolayer and spheroid growth conditions. Letrozole was an effective inhibitor of MCF-7aro monolayer cell proliferation, with an estimated 50% inhibitory concentration (IC50) of 50-100 nM, whereas an IC50 was not reached with anastrozole at any concentration tested (100-500 nM). An IC50 of tamoxifen was 1000 nM. Proliferation of T-47Daro monolayer cells was more sensitive to inhibition by all three agents; as with MCF-7aro cells, letrozole was the most effective inhibitor. MCF-7aro spheroids were slightly less sensitive than monolayer cells proliferation-inhibiting effects of letrozole (IC50 about 200 nM), and there was no significant inhibition with 100-200 nM anastrozole or 200 1000 nM tamoxifen. Letrozole and anastrozole significantly inhibited T-47Daro spheroid cell proliferation, at 15-25 and 50 nM, respectively, consistent with the greater sensitivity of T-47Daro monolayer cells to inhibition of proliferation by these agents. Tamoxifen failed to significantly inhibit T-47Daro spheroid cell proliferation over a 100-500 nM concentration range. Determination of aromatase inhibition in monolayers of both cell lines by a direct-access microsomal assay and an intact-cell assay revealed that letrozole was more active than anastrozole in monolayers of both cell lines and in both assays. In MCF-7aro spheroids following cell lysis, only letrozole significantly inhibited aromatase activity, supporting the conclusion that letrozole binds stronger to aromatase than anastrozole does. Our results demonstrate that MCF-7aro and T-47Daro spheroids could be a suitable model for evaluation of growth-inhibitory effects of agents used in hormonal therapy of breast cancer. PMID- 16263273 TI - A dispersion modelling approach to determine the odour impact of intensive poultry production units in Ireland. AB - The use of atmospheric dispersion modelling has become more common for the determination of odour impacts from existing poultry production facilities and the assessment of setback distances for new facilities. Setback distances for broiler, layer and turkey units were determined using the atmospheric dispersion model ISCST3 and the Environmental Protection Agency (EPA, Ireland) recommended criterion (C(98,1-h)6.0 ou(E) m(-3)) and a new odour annoyance criterion (C(98,1 h) 9.7 ou(E) m(-3)) developed in this study. For a typical size unit in Ireland, maximum setback distances of 660, 665 and 1035 m were calculated for 40,000 broilers, 40,000 layers and 10,000 turkeys respectively at the current limit (C(98,1-h) 6.0 ou(E) m(-3)). However, if the suggested odour impact criterion (C(98,1-h) 9.7 ou(E) m(-3)) is implemented, the maximum setback distances decrease to 460, 500 and 785 m for broilers, layers and turkeys, respectively. PMID- 16263274 TI - Production of pectinase from deseeded sunflower head by Aspergillus niger in submerged and solid-state conditions. AB - Studies were carried out on the production of pectinases using deseeded sunflower head by Aspergillus niger DMF 27 and DMF 45 in submerged fermentation (SmF) and solid-state fermentation (SSF). Higher titres of endo- and exo-pectinases were observed when medium was supplemented with carbon (4% glucose for SmF and 6% sucrose for SSF) and nitrogen (ammonium sulphate, 0.3% for both SmF and SSF) sources. Green gram husk proved to be relatively a better supplement to attain higher yield of endo-pectinase (11.7 U/g) and exo-pectinase (30.0 U/g) in solid state conditions. Maximum production of endo-pectinase (19.8 U/g) and exo pectinase (45.9 U/g) by DMF 45 were recorded in SSF when compared to endo pectinase (18.9 U/ml) and exo-pectinase (30.3 U/ml) by DMF 27 in SmF under optimum process conditions. PMID- 16263275 TI - Organic matter evolution during co-composting of the organic fraction of municipal waste and poultry manure. AB - The study concerned the evolution of organic matter, and the humification process, during the co-composting of the organic fraction of municipal solid waste (OMSW) and poultry manure (PM); the study was made with two different mixtures (OMSW:PM ratios of 3:2 and 2:3, wet weight:wet weight) and two different particle sizes (1 and 0.2cm). The results suggested that the composting process proceeded unhindered throughout the degradation of easily degradable materials like hemicellulose, and that of the rather less degradable cellulose and lipids, and the concentration of recalcitrant material, i.e. a ligno-humic (LU) fraction. These processes were more evident for mixtures with lower particle size. Throughout the composting, in all mixtures studied, humification proceeded by the formation of a new HA fraction, which was probably the result of the partial degradation and solubilization of more complex insoluble organic molecules, i.e. humin fraction. PMID- 16263276 TI - A concise asymmetric route for the synthesis of a novel class of glucocorticoid mimetics containing a trifluoromethyl-substituted alcohol. AB - An asymmetric route was developed for the synthesis of a class of novel glucocorticoid receptor ligand derivatives 1. The key step of this synthesis involves a diastereoselective addition of chiral sulfoxide anion to a trifluoromethyl ketone precursor. The resulting diastereomers are readily separable and can be converted to the corresponding chiral epoxide and chiral alkyne intermediates (2 and 3). This sequence of reactions is suitable for large scale preparation of these chiral intermediates and derivatives of 1. The absolute stereochemistry of the biologically active enantiomer of these GR ligands has also been determined. PMID- 16263277 TI - Computer-aided design of non-nucleoside inhibitors of HIV-1 reverse transcriptase. AB - Design principles are delineated for non-nucleoside inhibitors for HIV-1 reverse transcriptase (NNRTIs). Simultaneous optimization of binding affinity for wild type RT, tolerance for viral mutations, and physical properties is pursued. Automated lead generation with the growing program BOMB, Monte Carlo simulations with free-energy perturbation theory for lead optimization, and property analysis with QikProp are featured. An initial 30 microM lead has been optimized rapidly to the 10 nM level. PMID- 16263278 TI - Novel phenylamino acetamide derivatives as potent and selective kappa opioid receptor agonists. AB - A novel series of phenylamino acetamide derivatives was synthesized. These amides were shown to be potent and selective kappa opioid receptor agonists. PMID- 16263279 TI - A new chemical tool for exploring the role of the PDE4D isozyme in leukocyte function. AB - Nicotinamide (2) is a potent and selective inhibitor of the PDE4D isozyme and as a chemical tool selectively blocks eosinophil mediator release and chemotaxis thus linking the role of PDE4D to eosinophil function. PMID- 16263280 TI - Design, synthesis and in vitro antimalarial activity of an acylhydrazone library. AB - A library of acylhydrazone iron chelators was synthesized and tested for its ability to inhibit the growth of a chloroquine-resistant strain of Plasmodium falciparum. Some of these new compounds are significantly more active than desferrioxamine DFO, the iron chelator in widespread clinical use and also than the most effective chelators. PMID- 16263281 TI - Conformationally biased P3 amide replacements of beta-secretase inhibitors. AB - We have synthesized and evaluated a series of conformationally biased P3 amide replacements based on an isophthalamide lead structure. The studies resulted in the identification of the beta-secretase inhibitor 7m which has an in vitro IC(50)=35 nM. The synthesis and biological activities of these compounds are described. PMID- 16263283 TI - New bicyclic cannabinoid receptor-1 (CB1-R) antagonists. AB - A series of conformationally constrained bicyclic derivatives derived from SR141716 was prepared and evaluated as hCB(1)-R antagonists and inverse agonists. Optimization of the structure-activity relationships around the 2,6-dihydro pyrazolo[4,3-d]pyrimidin-7-one derivative 2a led to the identification of two compounds with oral activity in rodent feeding models (2h and 4a). Replacement of the PP group in 2h with other bicyclic groups resulted in a loss of binding affinity. PMID- 16263282 TI - Synthesis and pharmacological evaluation of second-generation phosphatidic acid derivatives as lysophosphatidic acid receptor ligands. AB - Short-chain phosphatidic acid derivatives, dioctanoyl glycerol pyrophosphate (DGPP 8:0, 1) and phosphatidic acid 8:0 (PA 8:0, 2), were previously identified as subtype-selective LPA(1) and LPA(3) receptor antagonists. Recently, we reported that the replacement of the phosphate headgroup by thiophosphate in a series of fatty alcohol phosphates (FAP) improves agonist as well as antagonist activities at LPA GPCR. Here, we report the synthesis of stereoisomers of PA 8:0 analogs and their biological evaluation at LPA GPCR, PPARgamma, and ATX. The results indicate that LPA receptors stereoselectively interact with glycerol backbone modified ligands. We observed entirely stereospecific responses by dioctyl PA 8:0 compounds, in which (R)-isomers were found to be agonists and (S) isomers were antagonists of LPA GPCR. From this series, we identified compound 13b as the most potent LPA(3) receptor subtype-selective agonist (EC(50)=3 nM), and 8b as a potent and selective LPA(3) receptor antagonist (K(i)=5 nM) and inhibitor of ATX (IC(50)=600 nM). Serinediamide phosphate 19b was identified as an LPA(3) receptor specific antagonist with no effect on LPA(1), LPA(2), and PPARgamma. PMID- 16263285 TI - Targeting the DNA minor groove with fused ring dicationic compounds: comparison of in silico screening and a high-resolution crystal structure. AB - The crystal structure of the DNA minor groove biphenyl benzimidazole diamidine ligand DB819 has been determined, bound to the DNA sequence d(CGCGAATTCGCG)(2), at a resolution of 1.36 Angstrom. Conditions for reliable in silico docking that reproduce the observed position of the ligand in the minor groove have been determined. PMID- 16263284 TI - Synthesis of functionalized 1,8-naphthyridinones and their evaluation as novel, orally active CB1 receptor inverse agonists. AB - Synthesis, SAR, and binding affinities are described for a new class of 1,8 naphthyridinone CB1 receptor specific inverse agonists. Food intake, knockout mouse, and pharmacokinetic evaluation of 14 indicate that this compound is an effective orally active modulator of CB1. PMID- 16263286 TI - An educational framework for triage nursing based on gatekeeping, timekeeping and decision-making processes. AB - INTRODUCTION: The role of the triage nurse has emerged in response to growing community demand for a more accessible and efficient emergency department (ED) service. The focus of triage research has been on measuring outcomes and improving the delivery of emergency care. This has meant that the context of care, and triage processes and practices have remained concealed. Thus, little evidence about the role and ways to prepare nurses for this role is available. The aim of this study was to provide insight and understanding needed to educate and support the triage nursing role in Australian EDs. METHODS: A 12-month ethnographic study of triage nursing practice was conducted in Sydney metropolitan EDs. Data were then collected from participant observation in four EDs and interviews with 10 triage nurses. Analysis used standard content and thematic analysis techniques. FINDINGS: Findings reveal that notions of timeliness, efficiency and equity are embedded in a culture of ED care. This sustains a particular cadence of care to which triage nurses are culturally oriented. Triage nurses maintain, negotiate and restore this cadence of emergency care by using gatekeeping, timekeeping and decision-making processes. CONCLUSION: The comprehensive study of triage nursing has led to the development of an educational framework based on the processes of gatekeeping, timekeeping and decision-making. PMID- 16263287 TI - Identification and characterisation of heel strike transient. AB - This study set out to ascertain the incidence of a heel strike transient (HST) and to verify whether modifications of muscle activation time could be correlated with the presence and the characteristic of a HST. Lower limb kinematics, kinetics and surface electromyography (SEMG) of normal subjects were examined during barefoot walking. A short-lived sharp HST was present in 76% of the trials while a smooth HST, of longer duration was seen in 13.3% of the trials. The transient phenomenon was absent in the remaining 10.7% of trials. The SEMG showed that alteration in the activation timing of muscles correlated with the presence of a HST. PMID- 16263288 TI - Understanding the structure-activity and structure-selectivity correlation of cyclic guanine derivatives as phosphodiesterase-5 inhibitors by molecular docking, CoMFA and CoMSIA analyses. AB - Molecular docking and 3D-QSAR analyses were performed to understand how PDE5 and PDE6 interact with a series of (49) cyclic guanine derivatives. Using the conformations of the compounds revealed by molecular docking, CoMFA and CoMSIA analyses resulted in the first quantitative structure-activity relationship (QSAR) and first quantitative structure-selectivity relationship (QSSR) models (with high cross-validated correlation coefficient q(2) and conventional correlation coefficient r(2) values) for predicting the inhibitory activity against PDE5 and the selectivity against PDE6. The high q(2) and r(2) values, along with further testing, indicate that the obtained 3D-QSAR and 3D-QSSR models will be valuable in predicting both the inhibitory activity and selectivity of cyclic guanine derivatives for these protein targets. A set of 3D contour plots drawn based on the 3D-QSAR and 3D-QSSR models reveal some useful clues to improve both the activity and selectivity by modifying structures of the compounds. It has been demonstrated that both the steric and electrostatic factors should appropriately be taken into account in future rational design and development of more active and more selective PDE5 inhibitors for the therapeutic treatment of erectile dysfunction (ED). PMID- 16263290 TI - Stereoselective synthesis and anti-inflammatory activities of 6- and 7-membered dioxacycloalkanes. AB - A class of 5-trifluoroacetylamino-1,3-dioxacycloalkanes, 5-benzoylamino-1,3 dioxacycloalkanes, and 5-amino-1,3-dioxacycloalkane compounds were stereoselectively synthesized as potential anti-inflammatory drug candidates. The anti-inflammatory activities of these compounds were tested using the xylene induced mouse ear edema model, from which multiple compounds possessing anti inflammatory properties which surpass aspirin were identified; these compounds were then compared to establish structure-activity relationships. PMID- 16263289 TI - Synthesis and biological evaluation of 9-(5',5'-difluoro-5' phosphonopentyl)guanine derivatives for PNP-inhibitors. AB - 9-(5',5'-Difluoro-5'-phosphonopentyl)guanine (DFPP-G) and its hypoxanthine analogue (DFPP-H) were modified by introducing a methyl group to all possible positions of the linker connecting a purine and difluoromethylenephosphonic acid moiety to evaluate the effects of the methyl group on inhibition against purine nucleoside phosphorylase. The methyl group on the linker affected the inhibition in a positional-dependent manner. Inhibitory potency of alpha-methyl and beta methyl-substituted analogues of DFPP-H increased by about 600- to 1000-fold upon converting to cyclopropane nucleotide analogue (+/-)-4. PMID- 16263291 TI - Design, synthesis, and biological activity of novel factor Xa inhibitors: improving metabolic stability by S1 and S4 ligand modification. AB - Serine protease factor xa (fXa) inhibitor 1 showed good ex vivo anti-fXa activity upon oral administration in rats. However, it has been revealed that 1 had low metabolic stability against human liver microsomes. To improve the metabolic stability, we attempted to modify the S1 and S4 ligands of 1. These modifications resulted in compound 34b, which exhibited selective anti-fXa activity and excellent anti-coagulation activity. PMID- 16263292 TI - Polyamine conjugates of meso-tritolylporphyrin and protoporphyrin IX: potential agents for photodynamic therapy of cancers. AB - An efficient five-step synthesis method was developed to obtain tritolylporphyrin and protoporphyrin IX polyamine conjugates. These compounds were composed of either one polyamine unit (spermidine or spermine) covalently tethered to monocarboxyphenyl tritolylporphyrin or two molecules of polyamines borne by protoporphyrin IX. In each compound, an aliphatic spacer arm is linked to the N(4) polyamine position. Photocytotoxicity of these new compounds was evaluated against K562 human chronic myelogenous leukemia cells and compared to Photofrin II; protoporphyrin IX polyamine conjugates exhibited much stronger photocytocicity than Photofrin II and were shown to readily induce necrosis in treated cells. PMID- 16263293 TI - Affinity prediction on A1 adenosine receptor agonists: the chemometric approach. AB - In this paper, we are presenting a quantitative-structure-activity relationship (QSAR) study performed on 21 selective A(1) adenosine receptor agonists plus the endogenous substrate, adenosine, so as to identify those predictors which play a key role in describing the binding of the ligand with the A(1) receptor. A large number of molecular descriptors plus a calculated receptor-agonist binding energy and atomic charges were taken into account to derive different QSAR models, using different regression techniques. The results obtained both with linear and nonlinear approaches converge to the selection of the same informative parameters, highlighting the correlation of these descriptors with the biological Response. The evaluation 'a priori' of these predictors could therefore represent a useful tool in the screening of large libraries of compounds and in the rational design of new selective agonists. PMID- 16263294 TI - Synthesis and biological activity of N,N-dialkylaminoalkyl-substituted bisindolyl and diphenyl pyrazolone derivatives. AB - New compounds, structurally related to the potent protein kinase C inhibitor staurosporine, with a bisindolylpyrazolone framework and substituted on the pyrazolone nitrogens with N,N-dialkylaminoalkyl side chain, were synthesized and evaluated for growth-inhibitory properties in several human cell lines. Many showed inhibition of TNF-alpha production in response to the tumor promotor TPA on HL-60 cells. The apoptotic activity on HeLa cells has been examined for several of these compounds. PMID- 16263295 TI - Theoretical study of structure, pKa, lipophilicity, solubility, absorption, and polar surface area of some centrally acting antihypertensives. AB - The methods of theoretical chemistry have been used to elucidate the molecular properties of the substituted imidazoline and oxazoline structures, a class of potent agonists and antagonists of imidazoline receptors. The geometries of various tautomers and isomers of 2-[2,6-dichlorophenylimino]imidazolidine (clonidine), 1-(N-dicyclopropylmethyl)amino-2-oxazoline (rilmenidine), 4-chloro-N (4,5-dihydro-1H-imidazol-2yl)-6-methoxy-2-methyl-5-pyrimidinamine (moxonidine), N (dicyclopropylmethyl)-4,5-dihydro-1H-pyrrol-2-amine (aminopyrroline), N dicyclopropylmethyl-4,5-dihydrothiazol-2-amine (aminothiazoline), 4,5-dihydro-2 (2-methoxyphenyl)-1H-imidazole (compound_6), 4,5-dihydro-2-(3-methylthiophen-2 yl)-1H-imidazole (compound_7), N-(2-chloro-4-iodophenyl)-4,5-dihydro-5-methyl-3H pyrrol-2-amine (LNP_911), N-amidino-3,5-diamino-6-chloropyrazine-carboxamide (amiloride), 2-(1,4-benzodioxan-2-yl)-2-imidazoline (idazoxan), (+/-)-2-(2-ethyl 2,3-dihydro-2-benzofuranyl)-2-imidazoline (efaroxan), (4-aminobutyl)guaninine (agmatine), and 1-methyl-9H-pyrido[3,4-b]indole (harmane) have been studied using Becke3LYP/6-31+G(d,p) and BP86/TZ2P DFT methods. The optimized geometries indicate that these molecules show a distinctly nonplanar configuration of the imidazoline and oxazoline moieties. In the gas-phase, rilmenidine and aminothiazoline exist in two forms (amino and imino), the amino tautomers being more stable by about 6 kJ/mol. The calculations showed, in agreement with experiments, that clonidine, moxonidine, and LNP_911 exist in a more stable imino tautomer. The tautomer containing the amino group is by about 30 kJ/mol less stable. Computations that include the effect of solvation indicated that also in water the relative stability order of individual tautomers (amino and imino forms) is preserved. The computed pKa values varied between 6.7 and 9.0, and correlate well with the available experimental pKa's found in the literature. Among the clinically useful antihypertensives moxonidine exhibits the lowest basicity in water. At pH = 7.4 only about 50% of this drug exists in ionized form. The available experimental partition coefficients of compounds investigated are best reproduced by the CLOGP method. The computed partition coefficients varied between -1.80 (agmatine) and 5.35 (LNP_911) (CLOGP). Clonidine, moxonidine, and rilmenidine are moderately lipophilic compounds with lipophilicities between these two extreme values. The computed solubilities (about 0.1-4 g/L) show that the imidazoline and oxazoline derivatives studied have very low water solubility. The analysis of molecular descriptors defined by Lipinski has shown that most of the compounds studied obey 'rule of five'. Amiloride and agmatine 'outlets' exhibit also the lowest absorption. Therefore, in the early stages of the design of ligands acting on imidazoline binding sites, it is becoming more important to determine the pKa, lipophilicity, water solubility, polar surface area, absorption, and other physicochemical properties associated with a drug, before synthetic work is undertaken, with the aim of avoiding the synthesis of compounds that are predicted to have poor biopharmaceutical characteristics. PMID- 16263296 TI - The 3D-QSAR analysis of 4(3H)-quinazolinone derivatives with dithiocarbamate side chains on thymidylate synthase. AB - Thymidylate synthase (TS) is a critical enzyme for DNA biosynthesis and many nonclassical lipophilic antifolates targeting this enzyme are quite efficient and encouraging as antitumor drug. In this paper, the binding model of 14 antifolates of 4(3H)-quinazolinone derivatives with dithiocarbamate side chains was examined using molecular simulation methods--FlexiDock and SCORE2.0. The resulted conformation and orientation of these antifolates were directly applied to CoMFA study. A good correlation between the calculated binding energies of these antifolates complexed with TS and their inhibitory activities was derived. The robust QSAR model, its three-dimensional contour map, and binding score for these antifolates derived from SCORE2.0 provided guidelines for structural optimization of current antifolates. PMID- 16263298 TI - Anti-HIV-1 protease activity of compounds from Boesenbergia pandurata. AB - Searching for anti-HIV-1 protease (PR) inhibitors of Thai medicinal plants led to the isolation of a new cyclohexenyl chalcone named panduratin C (1) and chalcone derivatives (2-6) from the methanol extract of Boesenbergia pandurata rhizomes. The known compounds were identified to be panduratin A (2), hydroxypanduratin A (3), helichrysetin (4), 2',4',6'-trihydroxyhydrochalcone (5), and uvangoletin (6). The structures of all compounds were elucidated on the basis of chemical and spectroscopic methods. It was found that 3 possessed the most potent anti-HIV-1 PR activity with an IC50 value of 5.6 microM, followed by 2 (IC50 = 18.7 microM), whereas other compounds exhibited only mild activity. Structure-activity relationships of these compounds on anti-HIV-1 PR activity are summarized as follows: (1) hydroxyl moiety at position 4 conferred higher activity than methoxyl group; (2) prenylation of dihydrochalcone was essential for activity; (3) hydroxylation at position 4''' reduced activity; and (4) introduction of double bond at C1' and C6' of chalcone gave higher activity. As regards active constituents contained in B. pandurata rhizomes, hydroxypanduratin A (3) and panduratin A (2) are active principles against HIV-1 PR. PMID- 16263297 TI - Synthesis and biological evaluation of new non-imidazole H3-receptor antagonists of the 2-aminobenzimidazole series. AB - A novel series of non-imidazole H(3)-receptor antagonists was developed, by chemical modification of a potent lead H(3)-antagonist composed by an imidazole ring connected through an alkyl spacer to a 2-aminobenzimidazole moiety (e.g., 2 [[3-[4(5)-imidazolyl]propyl]amino]benzimidazole), previously reported by our research group. We investigated whether the removal of the imidazole ring could allow retaining high affinity for the H(3)-receptor, thanks to the interactions undertaken by the 2-aminobenzimidazole moiety at the binding site. The imidazole ring of the lead was replaced by a basic piperidine or by a lipophilic p chlorophenoxy substituent, modulating the spacer length from three to eight methylene groups; moreover, the substituents were moved to the 5(6) position of the benzimidazole nucleus. Within both the 2-alkylaminobenzimidazole series and the 5(6)-alkoxy-2-aminobenzimidazole one, the greatest H(3)-receptor affinity was obtained for the piperidine-substituted compounds, while the presence of the p chlorophenoxy group resulted in a drop in affinity. The optimal chain length was different in the two series. Even if the new compounds did not reach the high receptor affinity shown by the imidazole-containing lead compound, it was possible to get good H(3)-antagonist potencies with 2-aminobenzimidazoles having a tertiary amino group at appropriate distance. PMID- 16263299 TI - The antioxidant activity of glucosamine hydrochloride in vitro. AB - The antioxidant potency of chitin derivative-glucosamine hydrochloride was investigated employing various established in vitro systems, such as superoxide (O2*-)/hydroxyl (*OH)-radical scavenging, reducing power, and ferrous ion chelating potency. As expected, we obtained several satisfying results, as follows: first, glucosamine hydrochloride had pronounced scavenging effect on superoxide radical. For example, the O2*- scavenging activity of glucosamine hydrochloride was 83.74% at 0.8 mg/mL. Second, the *OH scavenging activity of glucosamine hydrochloride was also strong and was about 54.89% at 3.2 mg/mL. Third, the reducing power of glucosamine hydrochloride was more pronounced. The reducing power of glucosamine hydrochloride was 0.632 at 0.75 mg/mL. However, ferrous ion-chelating potency was soft. Furthermore, ferrous ion-chelating potency, the scavenging rate of radical, and the reducing power of glucosamine hydrochloride increased with their increasing concentration, and they were concentration dependent. The multiple antioxidant activity of glucosamine hydrochloride was evident as it showed considerable reducing power, superoxide/hydroxyl-radical scavenging ability. These in vitro results suggest the possibility that glucosamine hydrochloride could be effectively employed as an ingredient in health or functional food, to alleviate oxidative stress. PMID- 16263300 TI - New substrates and inhibitors of gamma-aminobutyric acid aminotransferase containing bioisosteres of the carboxylic acid group: design, synthesis, and biological activity. AB - A series of potential substrates of gamma-aminobutyric acid aminotransferase (GABA-AT) with lipophilic bioisosteres of the carboxylic acid group (2-7) were synthesized and tested. Most of the synthesized compounds showed substrate activities with GABA-AT; 1H-tetrazole-5-propanamine (6) was the best of those tested. The potential time-dependent inhibitor of GABA-AT, 1H-tetrazole-5-(alpha vinyl-propanamine) (8), was designed based on the structures of 6 and the antiepilepsy drug vigabatrin (4-aminohex-5-enoic acid, 1). The synthesized compound 8 showed time-dependent inhibition of GABA-AT, but its potency is lower than that of vigabatrin. Methylation of the tetrazole group in 8 resulted in loss of time-dependent activity, suggesting that the tetrazole ring, the carboxylate bioisostere, exists in its deprotonated form in the enzyme active site. PMID- 16263301 TI - The development of 3D-QSAR study and recursive partitioning of heterocyclic quinone derivatives with antifungal activity. AB - It was reported that some 1,4-quinone derivatives such as 6-(N-arylamino)-7 chloro/6,7-bis[S-(aryl)thio]-5,8-quinolinedione and 6-arylthio-/5,6-arylamino-4,7 dioxobenzothiazoles have antifungal effects. To understand the structural basis for antifungal activity and guide in the design of more potent agents, we performed three-dimensional quantitative structure-activity relationship studies for a series of compounds using comparative molecular field analysis (CoMFA). The MIC values of 1,4-quinone derivatives on Aspergillus niger exhibited a strong correlation with steric and electrostatic factors of the 3D structure of molecules. The statistical results of the training set, cross-validated q(2) (0.683) and conventional r(2) (0.877) values, gave reliability to the prediction of inhibitory activity of a series of compounds. We also performed recursive partitioning (RP) analysis, used for the classification of molecules with activity using CART methods. Physicochemical, structural, and topological connectivity indices and E-state key descriptors were used for obtaining the decision tree models. The decision tree could classify the inhibitory activity of 1,4-quinone derivatives and its essential descriptors were S_aaN, Hbond donor, and Kappa-3. PMID- 16263302 TI - Synthesis and biological evaluation of a series of flavone derivatives as potential radioligands for imaging the multidrug resistance-associated protein 1 (ABCC1/MRP1). AB - Multidrug resistance (MDR) is one of the major problems affecting the treatment of cancer. In vivo visualization and quantification of MDR proteins would be of great value to better select the therapeutic strategy. Six flavone-based compounds were synthesized and evaluated for their cytotoxic activity and MDR reversing capacity using hMRP1 or hMDR1 overexpressing cell lines for in vitro assays. All the flavone derivatives were highly selective for hMRP1-expressing cell lines. These derivatives each used at 4muM (a non-cytotoxic concentration) enhance significantly the sensitivity of hMRP1-mediated MDR cell line toward doxorubicin toxicity. Their MDR-reversing capacity suggests that, in particular, the 4'-fluoroalkyloxy and 4'-iodo apigenin derivatives are potential new radiopharmaceuticals to visualize in vivo MRP1-mediated MDR phenomenon by PET or SPECT. PMID- 16263303 TI - Induction of apoptosis by photoexcited tetracyclic compounds derivatives of benzo[b]thiophenes and pyridines. AB - The antiproliferative activity, upon UVA irradiation, of two tetracyclic derivatives of benzo[b]thiophenes and pyridines, a benzo[b]thienopyridopyrimidone (1) and a thienocarboline (2), has been investigated in a panel of human tumor cell lines. The two compounds present a remarkable cytotoxicity after UVA irradiation (365 nm), reaching an IC50 of 0.1 microM in the leukaemia cell lines and 0.3-0.5 microM in the solid tumour cell lines. Their effect on the cell cycle was measured by flow cytometry in Jurkat cells. The compounds induce cell cycle perturbations and trigger a massive apoptosis as revealed by the externalisation of Annexin V-targeted residues at the outer plasmatic membrane. Furthermore the drugs induce, upon UVA irradiation significant variations of the mitochondrial potential (Deltapsi(mt)) measured by flow cytometry using the fluorochrome JC-1. In addition we characterized the mitochondrial production of reactive oxygen species (ROS) using the probe dihydroethidine (HE) and the oxidations of the mitochondrial phospholipid cardiolipin using the interacting probe nonyl acridine orange (NAO). Both compounds stimulate the production of ROS, and remarkably induce oxidation of cardiolipin. We have investigated the DNA-binding properties of these two compounds by means of UV-Vis spectroscopy and fluorescence. The two compounds exhibit a low affinity toward the macromolecule. The mode of binding was also investigated by means of flow linear dichroism (LD) which has revealed that the two compounds do not efficiently intercalate into DNA. Finally, the DNA photocleavaging properties of the test compounds were studied on pBR322 plasmid DNA as a model. Only compound 1 is able to induce a significant production of single strand breaks only after digestion with the base excision repair enzyme Endo III. Altogether these data suggest that DNA is not a preferential target of these molecules and other subcellular structures may be responsible for their high phototoxic activity. PMID- 16263304 TI - Interactions of tannic acid and its derivatives (ellagic and gallic acid) with calf thymus DNA and bovine serum albumin using spectroscopic method. AB - In the present investigation, an attempt has been made to study the interaction of chosen polyphenols (tannic, ellagic and gallic acids) with calf thymus DNA and bovine serum albumin (BSA) employing spectrofluorimetric technique. The fluorescence quenching of DNA-bound ethidium bromide (EB) and BSA-bound 1 anilinonaphthalene-8-sulfonic acid (ANS) by phenolic acids has been examined. As BSA contains two tryptophan residues, the polyphenols influence on protein by measuring the changes in the fluorescence of BSA in the presence of phenolic acids was also evaluated. Our experiments prove that there is a direct interaction between phenols and DNA or BSA. The obtained data suggest that used acids can intercalate to DNA and interact strongly with BSA. The strongest interactions were observed between DNA and ellagic acid and between BSA and tannic acid. The conformational changes were revealed in DNA and BSA after incubation with tested phenolic acids and the extent depended on the phenol structure and the used concentration. PMID- 16263305 TI - Effect of chlorine as substituent on the photochemistry and acid-base properties of beta-carboline alkaloids. AB - The UV-absorption, fluorescence excitation and emission spectra of the 6-chloro-, 8-chloro-, 6,8-dichloro-derivatives of nor-harmane, harmane and harmine and the 8 chloro-derivative of harmol were studied. These studies were performed in EtOH and in EtOH+1% perchloric acid solutions (pa). Furthermore, fluorescence quantum yields (phi(f)) in both media and in acetonitrile and acetonitrile + 1% perchloric acid solutions at 298 K were measured. The HOMO and LUMO energy, the positions (lambda(max)) and oscillator strength (f) of the 1S1 <-- 1S0 band for all the neutral and protonated beta-carbolines studied were calculated and compared with the experimental data. The pK(a) values in aqueous solution for for 6-chloro-, 8-chloro- and 6,8-dichloro-nor-harmane, harmane and harmine and 8 chloro-harmol were spectrophotometrically measured (pK(a(H2O)). The change of the acid-base character of these compounds on going from the ground state (pK(a)) to the first electronic excited singlet state (pKa*) as DeltapKa = pKa*-pKa, in ethanol solution at 298 K were calculated (DeltapK(a(EtOH))). Ground-state proton affinity (PA) for all the compounds studied defined as minus the enthalpy change of the reaction M + H(+) --> MH+ (gas state) were calculated. Basicity relative to pyridine (DeltaH(rPy)) defined as the enthalpy change of the isodesmic reaction MH(+) + Py --> M + PyH+ in gas state and in water solution, were also calculated (ab initio calculations). The effect of chlorine as substituent on the photochemistry and acid-base properties of the beta-carboline alkaloids is discussed. PMID- 16263306 TI - Pro- and anti-inflammatory response of acinar cells during acute pancreatitis. Effect of N-acetyl cysteine. AB - We investigate the ability of acinar cells to produce tumor necrosis factor-alpha (TNF-alpha) and interleukin-10 (IL-10) at different stages of acute pancreatitis (AP). Since oxidative stress is involved in the inflammatory response, the effect of N-acetyl cysteine (NAC) has also been evaluated. AP was induced in rats by bile-pancreatic duct obstruction (BPDO). NAC (50 mg/kg) was administered 1h before and 1h after BPDO. Acinar cells were incubated for 4 h at 37 degrees C in 5% CO2 atmosphere in absence and presence of 24-h BPDO-PAAF (20%, v/v) as stimulant agent. Acinar production of TNF-alpha and IL-10 was analysed by flow cytometry. Plasma amylase activity and histological studies of the pancreas indicated the severity of AP. PAAF significantly stimulated the acinar production of TNF-alpha and IL-10 in control rats. TNF-alpha production was also significantly stimulated in acinar cells of rats with AP, although a decrease in the pro-inflammatory response was found from 6 h after BPDO onwards. However, acinar cells failed to produce IL-10 from 3 h after BPDO. The protective effect of NAC treatment against oxidative cell damage reduced the pancreatic injury and maintained and enhanced the ability of acinar cells to produce IL-10 at early AP stages. As long as acinar cells were not severely damaged in the course of AP, greater ability to produce cytokines in response to PAAF was found in those with higher forward scatter (R2 cells). We suggest that the capability of acinar cells to maintain an appropriate balance between the production of pro- and anti inflammatory mediators could contribute to determine the degree of severity of AP. PMID- 16263307 TI - Laser flash photolysis of hydrogen peroxide to oxidize protein solvent-accessible residues on the microsecond timescale. AB - Footprinting of proteins by hydroxyl radicals generated on the millisecond to minute timescales to probe protein surfaces suffers from the uncertainty that radical reactions cause the protein to unfold, exposing residues that are protected in the native protein. To circumvent this possibility, we developed a method using a 248 nm KrF excimer laser to cleave hydrogen peroxide at low concentrations (15 mM, 0.04%), affording hydroxyl radicals that modify the protein in less than a microsecond. In the presence of a scavenger (20 mM glutamine), the radical lifetimes decrease to approximately 1 microsecond, yet the reaction timescales are sufficient to provide significant oxidation of the protein. These times are arguably faster than super-secondary protein structure can unfold as a result of the modification. The radical formation step takes place in a nanoliter flow cell so that only one laser pulse irradiates each bolus of sample. The oxidation sites are located using standard analytical proteomics, requiring less than a nanomole of protein. We tested the method with apomyoglobin and observed modifications in accord with solvent accessibility data obtained from the crystal structure of holomyoglobin. Additionally, the results indicate that the F-helix is conformationally flexible in apomyoglobin, in accord with NMR results. We also find that the binding pocket is resistant to modifications, indicating that the protein pocket closes in the absence of the heme group conclusions that cannot be drawn from current structural methods. When developed further, this method may enable the determination of protein-ligand interfaces, affinity constants, folding pathways, and regions of conformational flexibility. PMID- 16263308 TI - Close encounters of the first and second kind: T-DC and T-B interactions in the lymph node. AB - Cellular interactions in lymphoid organs initiate the immune response and determine its outcome. Using two-photon microscopy in the lymph node, several groups have begun to investigate the motility characteristics and interactions among T lymphocytes, B lymphocytes, and dendritic cells (DC) in lymphoid organs. In the first "close encounter", T cells of a particular antigen specificity interact with antigen-bearing dendritic cells and begin to activate. Activation of both CD4+ and CD8+ T cells evolves through several stages; from transient interactions to stable clusters and later to dissociation and proliferation of T cells (clonal expansion). The second "close encounter" requires that antigen engaged B cells become accessible to T cells by directed migration to the edge of the follicle. T cells and B cells then pair up and waltz together for an extended period, while helper T cells provide signals for B cells to differentiate into plasma cells. In this topical review, we compare the activation choreography of CD4+ T cells interacting first with dendritic cells, and then with B cells, during initiation of the humoral immune response. PMID- 16263309 TI - Complement consumption by Photobacterium damselae subsp. piscicida in seabream, red porgy and seabass normal and immune serum. Effect of the capsule on the bactericidal effect. AB - A virulent strain of Photobacterium damselae subsp. piscicida (Pdp) was grown without (C form) or with (C+ form) glucose supplementation, the latter to enhance capsule formation. Both forms were resistant to killing by normal serum of seabream, red porgy and seabass. However, the C form was killed by immune serum of all three fish species while the C+ form was killed only by seabream and red porgy sera and to a lesser extent than the C form. Both C and C+ forms consumed complement in normal serum and this consumption was enhanced by precoating the bacteria in specific fish antibody. Complement consumption was greatest in seabass serum, especially with antibody-coated C+ form yet in this case the bacteria were not killed. The killing of the C form in immune serum of all three fish species was completely inhibited by EGTA/Mg(2+), indicating that the mechanism of complement activation leading to killing of the bacteria was by the classical pathway. The results suggest that immune serum killing by the classical complement pathway may provide some degree of protection against pasteurellosis, but enhanced expression of the capsule by Pdp in vivo may restrict complement mediated killing, especially in immunised seabass. PMID- 16263310 TI - In vivo assessment of elbow flexor work and activation during stretch-shortening cycle tasks. AB - The purpose of this study was to use an electromyography (EMG) based muscle model to investigate the performance enhancement of stretch-shortening cycle (SSC) tasks at different elbow flexion-extension velocities. A torque motor was used to oscillate the forearms of seven healthy male subjects (23-40 years) during SSC and non-SSC contractions at four frequencies of movement (.58, 1.5, 2.4 and 3.3Hz) over a range of 105 degrees -162 degrees of elbow extension. The torque was integrated as a function of joint angle to yield the work produced by the elbow flexors. The elbow flexors were transcutaneously stimulated with a voltage equivalent to 60% maximum voluntary isometric contraction torque for 4s at 50Hz. EMG of the elbow flexors and extensors was recorded from the biceps and triceps respectively. The processed EMG was used to drive a Hill based model to predict the torque of the elbow flexors. Results indicate that muscle work increases from non-SSC to SSC trials. Work decreases for SSC and non-SSC trials with increasing velocity. The simulated constant activation muscle model predicted work well for all trials and conditions, indicating muscle model accuracy. The EMG driven model predicted well for all non-SSC trials, but significantly underestimated the work for SSC tasks, suggesting that the contractile component is directly involved in optimising muscle work during SSC tasks. PMID- 16263311 TI - Selective and interactive neural correlates of visual dimension changes and response changes. AB - In an event-related fMRI study, we investigated the neural correlates of visual dimension and response changes. We used a compound task, which required target selection by a singleton feature, a unique color or motion direction, before the appropriate motor response, which was determined by target orientation, could be selected. Both types of change elicited distinct patterns of activation, with dimension-change-related activation primarily in posterior visual areas and response-related activation primarily in motor-related areas of the parietal and frontal cortices. Response-change-related activation was delayed by about 1 s relative to dimension-change-related activation, suggesting that the latter is elicited by perceptual processes, whereas the former reflects response-related or post-response processes. Although dimension changes and response changes rely on different processes, they are not independent: response facilitation was observed for combined dimension and response repetitions, this facilitation, however, was disrupted by dimension changes. PMID- 16263312 TI - Sleep apnoea syndrome: a look at ethnicity, pathogeny and potential therapheutic approaches. PMID- 16263313 TI - Development of a metabolic network design and optimization framework incorporating implementation constraints: a succinate production case study. AB - We have developed a pathway design and optimization scheme that accommodates genetically and/or environmentally derived operational constraints. We express the full set of theoretically optimal pathways in terms of the underlying elementary flux modes and then examine the sensitivity of the optimal yield to a wide class of physiological perturbations. Though the scheme is general it is best appreciated in a concrete context: we here take succinate production as our model system. The scheme produces novel pathway designs and leads to the construction of optimal succinate production pathway networks. The model predictions compare very favorably with experimental observations. PMID- 16263315 TI - Successful treatment of multidrug-resistant Pseudomonas aeruginosa meningitis with intravenous and intrathecal colistin. PMID- 16263314 TI - Deoxyguanosine kinase mutations and combined deficiencies of the mitochondrial respiratory chain in patients with hepatic involvement. AB - The activity of deoxyguanosine kinase (DGUOK), a mitochondrial enzyme involved in the anabolism of mitochondrial (mt) deoxyribonucleotides, governs the maintenance of the mtDNA. Deleterious mutations of the DGUOK gene are thus associated with mtDNA depletion and result in combined deficiencies of mtDNA-encoded respiratory chain enzymes. With the aim to estimate the prevalence of DGUOK mutations in a cohort of 30 patients with hepatocerebral disease and combined respiratory chain deficiencies, we studied the DGUOK gene and identified previously unreported mutations in five families. Two patients and their affected sibs, born to non consanguineous parents, were homozygous for a missense mutation (M1T, and L250S, respectively). One patient presented a homozygous 4 pb insertion (796 insTGAT) and two other patients, and their affected sibs, were compound heterozygous (E165V/L266R and E211G/L266R, respectively). These findings allowed us to propose prenatal diagnosis in two families. In conclusion, we observed a high prevalence of DGUOK mutations (17%) in patients with hepatic involvement and combined respiratory chain deficiencies with hepatic involvement. PMID- 16263316 TI - Superantigen-induced multiple organ dysfunction in a toxin-concentration controlled and sequential parameter-monitored swine sepsis model. AB - OBJECTIVE: In order to examine the biological activity of low-dose and continuously infused superantigen, and to establish a superantigen-induced multiple organ dysfunction animal model, several pathophysiological parameters were sequentially monitored in a toxin-concentration-controlled pig model. METHODS: Anesthetized, mechanically ventilated and Swan-Ganz thermodilution catheter-inserted pigs were treated with toxic shock syndrome toxin-1 (TSST-1) by infusion at 2 microg/kg/h for 5 h. Monitoring was performed for both the infusion period and a subsequent 1-h post-infusion period. RESULTS: The serum concentration of TSST-1 was controlled so as to elevate it to a level over 1000 pg/mL within 1 h of initiation of infusion, and then gradually increased further and reached a plateau of about 2500 pg/mL at 4h after initiation. The animals showed a significant increase in cardiac output, the intrapulmonary arteriovenous shunt ratio, and infiltration of white blood cells into the lung. Although the observed increase in pulmonary vascular resistance was not statistically significant, it did correlate with the reduction in white blood cell counts. CONCLUSION: The superantigen TSST-1 plays an important role in the pathogenesis of Gram-positive bacterial sepsis by inducing multiple organ dysfunction. Thus, this model provides the first tool to allow the simultaneous examination of the serum toxin levels and other organ parameters in a time-course manner. PMID- 16263317 TI - Prevalence of brucellosis in the rural area of Kayseri, Central Anatolia, Turkey. PMID- 16263318 TI - A general ultrasound-assisted access to room-temperature ionic liquids. AB - The replacement of common organic solvents by room-temperature ionic liquids (RTILs) is a topical subject in both academia and industry. In the last decades, the number of applications for RTILs has followed an exponential curve and spilled over the boundaries of chemistry. Still, one of the main drawbacks of these compounds is their difficult access. The present ultrasound-assisted method affords a general and easy access to a large variety of room-temperature ionic liquids. PMID- 16263319 TI - Suicide by severing the arterio-venous subclavian dialysis catheter. AB - Haemodialysis access is an essential requirement for haemodialysis treatment in end-stage renal disease. The common forms are arteriovenous fistula (AVF) and arteriovenous grafts in ante-cubital fossa, forearm and upper thigh. Sometimes temporary or immediate access is created via a subclavian catheter or internal jugular catheter. This report is on a 79-year-old man who was suffering from chronic renal failure with a non-functional peripheral AVF; he was being dialysed through a permanent subclavian catheter and he became depressed due to continuing deterioration of his health. He used the easily accessible haemodialysis site as the method of suicide by cutting the tube that connected with the main vessel in his chest and bled to death. This highlights the requirement to assess carefully the patient's mental state in those on chronic haemodialysis, even though very few similar fatal cases have been previously reported. PMID- 16263320 TI - Forensic issues and possible mechanisms of sudden death in Rett syndrome. AB - A 20-year-old female with an established diagnosis of Rett syndrome was found dead in bed. There had been no history of recent deterioration in health and at autopsy no acute lesions were found. There was no evidence of trauma. Toxicological analysis of blood revealed therapeutic levels of carbamazepine and clonazepam. Death was attributed to the complications of Rett syndrome, an uncommon developmental disorder characterized by autistic type behaviour, hypotonia, stereotyped movements, seizures and growth failure, caused by mutations in the MECP2 gene on the X chromosome. Establishing the precise cause of sudden death in individuals with Rett syndrome may be difficult as epilepsy, defective autonomic nervous system control and cardiac arrhythmias may relate more to functional problems rather than to defects that can be demonstrated at autopsy. Thus, although there are a variety of well-documented underlying mechanisms that may cause sudden death in this condition, determining the exact sequence of events in an unwitnessed death may be more by inference and elimination, given the absence of pathognomonic and acute lethal lesions that are able to be found histopathologically. 'Complications of Rett syndrome' may, therefore, be the most accurate designation when individuals with this condition are found unexpectedly dead and no anatomical cause of death can be identified at autopsy. PMID- 16263321 TI - Death by attack from a wild boar. AB - Attacks on humans by wild animals causing fatal injuries are not uncommon in rural and forest areas of India. But death occurring due to attack by a wild boar is rare. As the victims of boar attack are usually recovered from dense forest areas, the investigating officers could be misled as to the nature of infliction of these multiple, fatal penetrating injuries to a possible homicide. Unlike the injuries inflicted by wild cats, canines and bulls, the hallmark of boar attack is the infliction of multiple penetrating injuries to the lower part of the body. This case is reported for its rarity, for the awareness of the possible injuries in such unnatural deaths, and for the factors predisposing to a boar attack. PMID- 16263322 TI - Increased gastric motility during 5-HT4 agonist therapy reduces response fluctuations in Parkinson's disease. AB - We investigated the clinical efficacy and tolerability of 45 mg/day mosapride, a selective 5-hydroxytryptamine type 4 (5-HT4) agonist, in an open-label study involving five patients with Parkinson's disease (PD) who had response fluctuations (RFs). 'On' time and motor function scores were determined, and gastric motility was measured by a radionuclide gastric emptying (GE) test, the most reliable quantitative method available. We found that mosapride therapy significantly shortened GE half-time, reduced RFs, and improved motor functions in all patients. There were no adverse reactions. We conclude that selective 5 HT4 agonist therapy is beneficial for patients with PD who have RFs. PMID- 16263323 TI - Active commuting to school among NSW primary school children: implications for public health. AB - Regular active commuting by walking/cycling can help maintain an active lifestyle. The frequency, duration and correlates of school active commuting were examined for primary school children in NSW, Australia. Walking/cycling-only and in combination with bus/car were of short duration (median 7 or 4 min, respectively) and their frequency dropped within a short distance (>0.75 km) from school. Apart from distance, child's age, school affiliation and perceived safety, regular walking/cycling (10 trips, 22%) was associated with parents' travel mode to work and with father taking the child to school. Frequent walking/cycling (5 trips, 37%) was associated with child's level of independence and the perceived benefits of active commuting. Behaviour change in this setting requires multi-level strategies. PMID- 16263324 TI - Detection of a raft-located estrogen receptor-like protein distinct from ER alpha. AB - 17Beta-estradiol (17beta-E2) elicits at the cell membrane rapid actions that remain insensitive to the inhibitory effect of ICI 182,780, a pure estrogen antagonist, and therefore cannot be attributed to the classic nuclear receptors. We addressed the question of the identity of the protein involved in these rapid actions. We first examined the responses of several cell lines for intracellular calcium mobilization, an effect not inhibited by ICI 182,780, tamoxifen and raloxifen. We then demonstrated the presence of binding sites in the membranes, by incubating them with antibodies directed against different domains of ER alpha, and by flow cytometry analysis. The membrane proteins were eluted by affinity chromatography using E2 conjugated to bovine serum albumin as a ligand. Western blots of the elution fractions using an antibody directed against the ligand binding site of ER alpha showed the existence of a protein of approximately 50 kDa. The protein was concentrated in the lipid rafts, together with another heavier form of approximately 66 kDa. The 50 kDa protein was immunoprecipitable, and co-immunoprecipitation experiments showed that it was associated with the Gbeta(1-4) protein, but not with caveolin-1. The protein was expressed in ER alpha-null cells, like HO-23 and Cos-7 cells. Therefore, in the lipid rafts, there exists a protein, similar to, but molecularly distinct from ER alpha. PMID- 16263325 TI - Level set based cerebral vasculature segmentation and diameter quantification in CT angiography. AB - A level set based method is presented for cerebral vascular tree segmentation from computed tomography angiography (CTA) data. The method starts with bone masking by registering a contrast enhanced scan with a low-dose mask scan in which the bone has been segmented. Then an estimate of the background and vessel intensity distributions is made based on the intensity histogram which is used to steer the level set to capture the vessel boundaries. The relevant parameters of the level set evolution are optimized using a training set. The method is validated by a diameter quantification study which is carried out on phantom data, representing ground truth, and 10 patient data sets. The results are compared to manually obtained measurements by two expert observers. In the phantom study, the method achieves similar accuracy as the observers, but is unbiased whereas the observers are biased, i.e., the results are 0.00+/-0.23 vs. 0.32+/-0.23 mm. Also, the method's reproducibility is slightly better than the inter-and intra-observer variability. In the patient study, the method is in agreement with the observers and also, the method's reproducibility -0.04+/-0.17 mm is similar to the inter-observer variability 0.06+/-0.17 mm. Since the method achieves comparable accuracy and reproducibility as the observers, and since the method achieves better performance than the observers with respect to ground truth, we conclude that the level set based vessel segmentation is a promising method for automated and accurate CTA diameter quantification. PMID- 16263326 TI - Health-related quality of life measured by the UW-QoL--reference values from a general dental practice. AB - The objective of this study was to obtain age and sex-specific reference values for the University of Washington head and neck cancer questionnaire version 4 (UW QoLv4) and to compare this with patients with oral and oropharyngeal cancer. Cross-sectional reference data was collected from 372 patients in six local general dental practices, 349 of whom presented for routine appointments. Quota sampling was used to collect data for similar numbers of patients by gender by four age bands (40-49, 50-59, 60-69, 70-79 yr). The longitudinal sample consisted of 450 consecutive patients undergoing primary surgery for previously untreated oral and oropharyngeal squamous cell carcinoma presenting to the Regional Maxillofacial Unit Liverpool, between the years 1995 and 2002. At baseline the key differences were anxiety, pain, swallowing, chewing, and mood. At 1yr there were big differences in all domains with deterioration in the oral cancer group. The difference was least notable in pain, shoulder, mood and anxiety. Reference data from a non-cancer population is very important when considering UW-QoL domains as an outcome parameter in clinical trials and also when discussing health-related quality of life outcomes with patients and their families. PMID- 16263327 TI - Integration of a handheld based anaesthesia rounding system into an anaesthesia information management system. AB - BACKGROUND: At the University Hospital Giessen, an anesthesia information management system (AIMS) is used for online record keeping of perioperative patient care, but preoperative anaesthesia assessments were still being recorded on paper and subsequently entered into the AIMS. Personal digital assistants (PDAs) seem to be useful instruments to establish a seamless digital anesthesiological documentation. OBJECTIVES: We decided to implement a solution for direct integration of data gathered during the preoperative assessment into the existing data management infrastructure. Parallel to the development of the system, we surveyed the future users to match their wishes and needs as far as possible. SYSTEM DESCRIPTION: A C program embedding the preoperative AIMS' data fields was developed. Data alignment with the Hospital information system (HIS) is controlled by a Java desktop software. The anaesthesiologist completes the available fields at the patient's bedside following the same algorithm and integrity check as the PC version. STATUS REPORT: Overall, 68% of the surveyed physicians supported the implementation of the system. The PDA solution has been available since May 2002. Data replication into the handheld and integration of mobile collected data into the AIMS generally work without problems. The HIS interconnection software converts the PDA file into the AIMS format for further processing. DISCUSSION: The preoperative anaesthetic assessment is a standardised task well suitable for conversion to an electronic data storage medium. Changing from redundant data entry in the OR to direct electronic recording at the patient's bedside seems simply logical. Handheld computers are inexpensive, flexible gadgets to realize this. PMID- 16263328 TI - Microbiological validation of smear microscopy after sputum digestion with bleach; a step closer to a one-stop diagnosis of pulmonary tuberculosis. AB - BACKGROUND: Smear microscopy is relatively insensitive for the diagnosis of TB. The digestion of sputum with household bleach prior to smear preparation has been reported to improve its sensitivity. This method has not been validated. METHODS: Seven hundred and fifty six patients with symptoms suggestive of pulmonary TB (PTB) were asked to submit 3 sputum specimens for direct microscopy. One specimen was selected at random for culture and another specimen was digested to prepare a further smear. The WHO case definition (>or=2 positive smears or one positive smear and positive culture) was used to compare the sensitivity and specificity of the smears. FINDINGS: Four hundred and fifty five (60%) patients were culture positive. Of these, 235 (31%) had "definite" PTB and 223 (29%) "very likely" PTB (smear-negative, culture-positive). The WHO case definition identified 51% (235/458) of the patients with "definite" or "very likely" PTB. One digested smear detected 219 (93%) of the 235 patients with "definite" PTB and 10 patients with "very likely" PTB (sensitivity (95%CI) 50% (45-55%); specificity 99% (97 100%)). The positive and negative predictive values for one digested smear were 98% (95-99%) and 56% (52-60%) respectively, which were not different (p>0.5) to the WHO case definition (100% and 57%, respectively). INTERPRETATION: One bleach digested smear is as sensitive and specific as the WHO case definition for the diagnosis of PTB. PMID- 16263330 TI - Recent innovations in colposcopy practice. AB - Colposcopic practice continues to evolve. As its need has expanded, so has the role of training, audit and continuing medical education. The recently published National Health Service Cervical Screening Programme clinical guidelines document covers almost every aspect of clinical practice in an evidence-based directory. Excision of the transformation zone (TZ) may be a very minor or major entity. The recent TZ classification system of the International Federation of Cervical Pathology and Colposcopy attempts to clarify and standardize nomenclature so that therapy can be realistically compared. The role of human papillomavirus (HPV) in clinical practice continues to be controversial and has not yet found a place in the UK. For the evaluation of borderline nuclear abnormal smear and for post treatment surveillance, HPV is clinically useful and efficient. Other biological tumour markers are likely to become clinically useful as their predictive profiles emerge. PMID- 16263329 TI - Interactions of anti-sigma factor antagonists of Mycobacterium tuberculosis in the yeast two-hybrid system. AB - Anti-sigma factor antagonists (anti-anti-sigma factors) play critical roles in regulating the expression of alternative sigma factors in response to specific stress signals. The Clusters of Orthologous Groups (COG) database has identified the existence of six genes, Rv0516c, Rv1364c, Rv1365c, Rv1904, Rv2638 and Rv3687c (grouped under the cluster COG1366), encoding potential anti-sigma factor antagonists in Mycobacterium tuberculosis. These molecules are speculated to regulate the expression of sigma factor SigF of M. tuberculosis in response to stress signals. Since signaling occurs via physical interactions of proteins (protein-protein interaction), we investigated whether the anti-sigma factor antagonists of M. tuberculosis interact with anti-sigma factor RsbW (Rv3287c) or the sigma factor SigF (Rv3286c) in the yeast two-hybrid system. The results revealed that most of the anti-sigma factor antagonists interact with either RsbW or SigF or both. In addition, some anti-sigma factor antagonists also displayed limited interactions between themselves. These interactions suggest that they possibly transduce some signals to SigF and between themselves. PMID- 16263331 TI - Choroidal neovascularization in transgenic mice expressing prokineticin 1: an animal model for age-related macular degeneration. AB - The prognosis of choroidal neovascularization (CNV) in age-related macular degeneration (AMD) is poor and existing treatments are limited in retarding the progression of disease. The development of an animal model for AMD will be beneficial for finding potential treatments, including gene therapy. Recently prokineticin 1 (hPK1) was identified as a mitogen of fenestrated endothelium. We hypothesized that hPK1 could induce CNV, a hallmark of the exudative or wet form of AMD, since the endothelium of the choriocapillaris, but not retinal endothelium, has fenestration. We generated transgenic mice expressing hPK1 in the retina using the rhodopsin promoter. In these transgenic mice, an enlarged vascular bed of choroid resembling CNV was observed without any morphological changes in the retinal vasculature. In addition, the major fluorophore of lipofuscin, N-retinylidene-N-retinylethanolamine, which has several potential cytotoxic effects on the RPE, was accumulated approximately twice as much in the transgenic mouse eyes compared to controls. hPK1 could be one of the causative factors of AMD and the transgenic mouse exhibiting CNV may be useful to establish treatments for the wet form of AMD. PMID- 16263332 TI - Identification of mouse AAV capsid-specific CD8+ T cell epitopes. AB - Adeno-associated virus has been developed for use as a gene transfer vector. To understand the impact of AAV capsid-specific CD8(+) T cells on AAV-mediated gene transfer, we identified CD8(+) T cell epitopes for AAV-2 and AAV-8 capsid in C57BL/6 (H-2(b) MHC haplotype) and BALB/c (H-2(d) MHC haplotype) mice. Mice of both the H-2(b) and the H-2(d) haplotypes recognized epitopes on AAV-2 and AAV-8 capsid. T cells from H-2(b) mice recognized an epitope that was conserved between AAV-2 and AAV-8 capsid. Cross-reactivity of AAV-specific CD8(+) T cells induced by different AAV serotypes may have important implications for gene transfer. Identification of these epitopes will facilitate studies of immune response to AAV capsid in mouse models. PMID- 16263333 TI - Acute thrombosis of a prosthetic mitral valve. AB - We report the case of a patient who was transferred to our hospital with acute thrombosis of a prosthetic mitral valve. Her admission INR was subtherapeutic. The transoesophageal echocardiographic images are presented. The patient underwent urgent reoperation and made a good recovery. PMID- 16263334 TI - Hazards of percutaneous PFO closure. PMID- 16263335 TI - Cognitive and behavioral effects of nocturnal epileptiform discharges in children with benign childhood epilepsy with centrotemporal spikes. AB - This review addresses the effects of subclinical localized epileptiform discharges during sleep in children with benign childhood epilepsy with centrotemporal spikes (BCECTS) on cognition and behavior. A diversity of educational deficits, behavioral impairments, language delay, and neuropsychological test results have been reported in children with BCECTS. The occurrence of atypical seizure characteristics seems to be related to language delay, and several atypical EEG characteristics are related to cognitive or behavioral problems. It remains to be shown whether treatment of nocturnal discharges in children with BCECTS is indicated to improve cognitive and behavioral problems. PMID- 16263336 TI - Elementary school epilepsy survey (ESES): a new measure of elementary school students' knowledge and attitudes about epilepsy. AB - OBJECTIVE: No peer-reviewed, published, psychometrically tested scales are available to assess elementary school students' general knowledge and attitudes about epilepsy. Such a scale is needed for evaluation of the effectiveness of classroom education programs. The purpose of this work was to develop and validate a brief, reliable scale for grades 4-6 to assess students' knowledge and attitudes about epilepsy and persons diagnosed with epilepsy. METHODS: Development of the 22-item Elementary School Epilepsy Survey (ESES) followed standard protocol for scale development. It includes a 12-item Knowledge subscale and a 10-item Attitudes subscale. The ESES was administered during regular classroom time given and repeated 1 week later. No educational intervention took place. RESULTS: Mean age of the 155 students was 11 years (range 9.8-13.9): grade 4, 56 students; grade 5, 36; and grade 6, 63 students. The ESES Total scale and Attitudes subscale had good internal consistency (Cronbach's alpha=0.72 and 0.81, respectively). As expected, the Knowledge subscale had low internal reliability (Cronbach's alpha=0.50). Test-retest scores indicated good reliability and strong discriminant validity, with significant increases noted in all ESES scores with increasing age and in those who knew someone with epilepsy. CONCLUSION: The ESES detects developmental and experiential trends in students' knowledge and attitude about epilepsy. It has good internal consistency and test-retest reliability. PMID- 16263337 TI - Introduction to JAK/STAT signaling and the vasculature. PMID- 16263338 TI - The development of a reagentless lactate biosensor based on a novel conducting polymer. AB - A reagentless lactate biosensor is described, based on an electropolymerized copolymer film poly(5-hydroxy-1,4-naphthoquinone-co-5-hydroxy-3-acetic acid-1,4 naphthoquinone). The quinone group, as part of the polymer backbone, is electroactive and very stable in neutral aqueous medium. It can therefore act as an immobilized mediator for the enzyme recycling, at a working potential much lower than those commonly reported in the literature for other mediators. Experimental conditions for amperometric measurements (temperature, pH) are studied, especially the interference between quinone and molecular oxygen to investigate the enzyme/quinone recycling kinetic. Some well-known interferents are shown to have no measurable effect on the amperometric curves. PMID- 16263339 TI - Flies and their golden apples: the effect of dietary restriction on Drosophila aging and age-dependent gene expression. AB - Reduced nutrient availability (dietary restriction) extends lifespan in species as diverse as yeast, nematode worms, Daphnia, Drosophila, and mammals. Recent demographic experiments have shown that moderate nutrient manipulation in adult Drosophila affects current mortality rate in a completely reversible manner, which suggests that dietary restriction in Drosophila increases lifespan through a reduction of the current risk of death rather than a slowing of aging-related damage. When examined in the light of the new demographic data, age-dependent changes in gene expression in normal and diet-restricted flies can provide unique insight into the biological processes affected by aging and may help identify molecular pathways that regulate it. PMID- 16263340 TI - DNA polymerase iota-dependent translesion replication of uracil containing cyclobutane pyrimidine dimers. AB - Analysis of the spectrum of UV-induced mutations generated in synchronized wild type S-phase cells reveals that only approximately 25% of mutations occur at thymine (T), whilst 75% are targeted to cytosine (C). The mutational spectra changes dramatically in XP-V cells, devoid of poleta, where approximately 45% of mutations occur at Ts and approximately 55% at Cs. At the present time, it is unclear whether the C-->T mutations actually represent true misincorporations opposite C, or perhaps occur as the result of the correct incorporation of adenine (A) opposite a C in a UV-photoproduct that had undergone deamination to uracil (U). In order to assess the role that human poliota might play, if any, in the replicative bypass of such UV-photoproducts, we have analyzed the efficiency and fidelity of pol iota-dependent bypass of a T-U cyclobutane pyrimidine dimer (CPD) in vitro. Interestingly, pol iota-dependent bypass of a T-U CPD occurs more efficiently than that of a corresponding T-T CPD. Guanine (G) was misincorporated opposite the 3'U of the T-U CPD only two-fold less frequently than the correct Watson-Crick base, A. While pol iota generally extended the G:3'U-CPD mispairs less efficiently than the correctly paired primer, pol iota-dependent extension was equal to, or greater than that observed with human pols eta and kappa and S. cerevisiae pol zeta under the same assay conditions. Thus, we hypothesize that the ability of pol iota to bypass T-U CPDs through the frequent misincorporation of G opposite the 3'U of the CPD, may provide a mechanism whereby human cells can decrease the mutagenic potential of these lesions. PMID- 16263341 TI - Rapid high-performance liquid chromatographic method for Vitamin C determination in human milk versus an enzymatic method. AB - Vitamin C is an antioxidant that can be considered a possible biomarker of oxidative stability in human milk. A high-performance liquid chromatographic method was developed and validated for determining the total Vitamin C (ascorbic acid and dehydroascorbic acid) and ascorbic acid levels in human milk. This method was then compared with an enzymatic method (a Colorimetric technique) for quantifying ascorbic acid levels. Repeatability and reproducibility were acceptable for all methods. However, the high-performance liquid chromatography (HPLC) technique provided more satisfactory results than the enzymatic method due to this last method detected 37% less ascorbic acid and does not determine the total Vitamin C because of the enzymatic method cannot reduce the dehydroascorbic acid (DHA) to ascorbic acid. Furthermore, the HPLC method has the added advantages that it requires less reagents and material, and is simpler and less time consuming than the enzymatic method. In conclusion, the drawbacks of this enzymatic method would justify its substitution for a HPLC method. PMID- 16263342 TI - Direct determination of glucuronide and sulfate of p-hydroxymethamphetamine in methamphetamine users' urine. AB - Two conjugates of p-hydroxymethamphetamine (p-OHMA), p-OHMA-glucuronide (p-OHMA Glu) and p-OHMA-sulfate (p-OHMA-Sul) have been identified in methamphetamine (MA) users' urine by using liquid chromatography-high resolution tandem mass spectrometry (LC-HRMS-MS). The synthesis of p-OHMA-Glu and p-OHMA-Sul, and an LC MS procedure for the simultaneous determination of MA and its four metabolites, amphetamine (AP), p-OHMA, p-OHMA-Glu and p-OHMA-Sul, in urine have also been established. After deproteinizing urine samples with methanol, LC-MS employing a C(18) semi-micro column with a gradient elution program provided the successful separations and MS determinations of these analytes within 20 min. Based on the established method, p-OHMA-Sul was detected at higher concentrations than p-OHMA Glu in all of the three urine samples tested. These data suggest that sulfation is a major pathway in the MA phase II metabolism. PMID- 16263343 TI - Self-perceived normality in defecation habits. AB - BACKGROUND: Available information on normal bowel habits was mainly gathered by means of telephone interviews or mailed questionnaires. AIMS: We undertook a prospective study to evaluate the defecatory habits in subjects perceiving themselves as normal concerning this function. SUBJECTS AND METHODS: A questionnaire (4-week diary with "yes-no" daily answers to six questions concerning bowel habits) was distributed to 204 subjects perceiving their defecation behaviour as normal. RESULTS: The completed questionnaire was returned by 140 subjects. No significant differences were found between sexes or age groups for any variable, even though straining at stool and feeling of incomplete and/or difficult evacuation showed a trend to increase with age. No subject had less than three bowel movements per week or more than three per day. The percentage of symptoms linked to an abnormal defecatory behaviour was well below 10%. Fifty-five percent of subjects reported at least one parameter of abnormal functioning; the most frequent was straining at stool and the rarer was the manual manoeuvres to help defecation. CONCLUSIONS: In normal subjects the prevalence of symptoms considered in Rome II criteria as part of an abnormal defecatory behaviour (in more than 25% of defecations) is well below 10%, manual manoeuvres are almost never used to help defecation, and the frequency of defecations is at least three per week. PMID- 16263344 TI - Identical endoscopic and histological finding on two monozygotic twins affected by coeliac disease. PMID- 16263346 TI - Innate immunity in human bone. AB - Bone has exceptional regenerative properties. Oral bone appears to be particularly resistant to infection despite exposure to oral flora, even in circumstances such as oral surgery where the thin mucosal layer covering the bone is disrupted. The goal of this study was to determine whether the innate immune system of antimicrobial peptides exists inside bone. Biopsies of non-infected and chronically infected mandibular bone were harvested from patients during maxillofacial surgical procedures. Bone biopsies from the iliac crest and fibula served as controls. Immunohistochemical staining was performed, directed against the human beta-defensin antimicrobial peptides (hBD) -1, -2 and -3. In addition, cultures of osteoblast-like cells were examined for the presence of each of the three beta-defensins and their mRNA transcripts. All three human beta-defensins were detected within the mineralized bone matrix of chronically infected mandibular bone in the vicinity of the endosteum and osteocytes. hBD-1, -2 and -3 were also found in the cytoplasm of osteocytes. Expression of all three beta defensins was detected in each of the non-infected bone types including the controls, however, to a lesser degree than that found in the chronically infected mandibular bone. This may reflect upregulation of antimicrobial peptide expression in the presence of chronic infection. Cultures of non-infected osteoblast-like cells were found to express mRNA for each of hBD-1, -2 and -3. Immunohistochemical staining of the cultures was positive for hBD-1 and -2, but not for hBD-3. We provide the first evidence of a previously unrecognized innate immunological function of bone through the demonstration of the presence of the human beta-defensins hBD-1, -2 and -3 in bone. PMID- 16263347 TI - Molecular cloning, expression and immunolocalization of a novel human cementum derived protein (CP-23). AB - Cementum is a unique mineralized connective tissue that covers the root surfaces of the teeth. The cementum is critical for appropriate maturation of the periodontium, both during development as well as that associated with regeneration of periodontal tissues, IU; however, one major impediment to understand the molecular mechanisms that regulate periodontal regeneration is the lack of cementum markers. Here we report on the identification and characterization of one such differentially human expressed gene, termed "cementum protein-23" (CP-23) that appears to be periodontal ligament and cementum-specific. We screened human cementum tumor-derived cDNA libraries by transient expression in COS-7 cells and "panning" with a rabbit polyclonal antibody against a cementoblastoma conditioned media-derived protein (CP). One isolated cDNA, CP-23, was expressed in E. coli and polyclonal antibodies against the recombinant human CP-23 were produced. Expression of CP-23 protein by cells of the periodontium was examined by Northern blot and in situ hybridization. Expression of CP-23 transcripts in human cementoblastoma-derived cells, periodontal ligament cells, human gingival fibroblasts and alveolar bone-derived cells was determined by RT-PCR. Our results show that we have isolated a 1374-bp human cDNA containing an open reading frame that encodes a polypeptide with 247 amino acid residues, with a predicted molecular mass of 25.9 kDa that represents CP species. The recombinant human CP-23 protein cross-reacted with antibodies against CP and type X collagen. Immunoscreening of human periodontal tissues revealed that CP-23 gene product is localized to the cementoid matrix of cementum and cementoblasts throughout the entire surface of the root, cell subpopulations of the periodontal ligament as well as cells located paravascularly to the blood vessels into the periodontal ligament. Furthermore, 98% of putative cementoblasts and 15% of periodontal ligament cells cultured in vitro expressed CP-23 gene product. Cementoblastoma cells and periodontal ligament cells contained a 5.0 kb CP-23 mRNA. In situ hybridization showed strong expression of CP-23 mRNA on cementoblast, cell subpopulations of the periodontal ligament and cells located around blood vessels into the periodontal ligament. Our results demonstrate that CP-23 represents a novel, tissue-specific-gene product being expressed by periodontal ligament subpopulations and cementoblasts. These findings offer the possibility to determine the cellular and molecular events that regulate the cementogenesis process during root development. Furthermore, it might provide new venues for the design of translational studies aimed at achieving predictable new cementogenesis and regeneration of the periodontal tissues. PMID- 16263349 TI - Short- and long-term outcomes of the titanium-NO stent registry. AB - BACKGROUND: Five to 15% of the population have allergy to nickel, chromium, or molybdenum, which is a potential cause for in-stent restenosis. The Titan stent is made of stainless steel and is coated with titanium-nitride oxide (TiNOX), which completely prevents the discharge of metal elements. We performed a real life multicenter registry to assess the short- and long-term characteristics of the Titan stent. METHODS AND RESULTS: A total of 103 Titan stents was implanted in 100 patients. Patients were 61.4+/-12.6 years old (81 men). Risk factors included hypercholesterolemia (63%), hypertension (53%), diabetes mellitus (DM; 35%), and current smoking (23%). Indications for PCI (percutaneous coronary intervention) were acute coronary syndromes (ACS) in 68% [acute ST elevation myocardial infarction (MI) in 8%], stable AP (angina pectoris) in 25%, and silent ischemia in 7% of the patients. Fifty-two percent of the treated lesions were of Type B2 or C. Lesion length was 14.3+/-2.9 mm and stent diameter was 3.06+/-0.36 mm. Indications for stenting were prevention of restenosis in 66%, residual stenosis in 33%, dissection in 13%, acute MI in 13%, and in-stent restenosis in 7% of the patients. Procedural success was 100%, with no complications. At 30 days, there were no major adverse cardiac events (MACE), including death, MI, and revascularization. At 180 days, only three patients had TVR (target vessel revascularization); two had TLR (target lesion revascularization) (one PCI and one CABG [coronary artery bypass grafting]), and one patient had a new narrowing proximal to the stent and underwent CABG due to multivessel disease. CONCLUSIONS: The Titan stent has a remarkable safety profile in high-risk patients and complex coronary lesions and excellent short- and long-term outcome with a very low clinical TLR rate. PMID- 16263350 TI - Intracoronary beta-irradiation prevents excessive in-stent neointimal proliferation in de novo lesions of patients with high plasma ACE levels. The BetAce randomized trial. AB - BACKGROUND: This study evaluated vascular brachytherapy (VBT) as a potent antiproliferative treatment to prevent in-stent restenosis (ISR) after coronary angioplasty of de novo lesions in patients carrying the D allele of the I/D polymorphism of the ACE gene and high ACE plasma levels (>34 U/l). METHODS AND MATERIALS: A prospective randomized trial was designed to detect a 30% improvement in the minimal lumen diameter (MLD) of the stenotic artery, as measured by quantitative coronary analysis (QCA), 6 months following VBT at the time of stented angioplasty. All patients were carriers of the D allele of the ACE gene, with plasma ACE levels >34 U/l. RESULTS: Thirty-one patients (33 stenoses) were allocated to stent implantation (control group) and 30 patients (31 stenoses) to VBT and stented angioplasty. After angioplasty, in-stent MLD was similar in the two groups. At 6 months in the control group, in-stent MLD had decreased to 1.74+/-0.8 versus 2.25+/-1.05 mm in the VBT group (P=.04). The mean in-stent diameter was 2.3+/-0.8 mm in the control group versus 2.9+/-1.05 mm after VBT (P=.02), and the restenosis rate was 37.5% versus 17.9%, respectively (P=.08). At 6 months, a higher need for target vessel revascularization (TVR) was observed in the control group: 35.5% versus 13.3% (P=.04). CONCLUSIONS: This randomized study confirms that patients with high plasma ACE concentrations are exposed to an increased risk for ISR after coronary stenting. The preventive use of VBT in these patients reduced neointimal formation by 65% such that the MLD at follow-up was increased by 29% compared with the control group. PMID- 16263351 TI - Potential of an intracardiac electrogram for the rapid detection of coronary artery occlusion. AB - BACKGROUND: Early identification of acute MI and prompt intervention can improve clinical outcomes. It would be valuable to identify a method that could allow the earliest possible detection of myocardial injury or ischemia. METHODS AND RESULTS: This article reports one of the first clinical investigations to examine the ability of an intracardiac right ventricular (RV) electrode to identify the early onset of myocardial ischemia/injury in a cohort of patients undergoing balloon occlusion of a coronary artery during percutaneous transluminal coronary angioplasty. The primary data set for analysis included observations from 14 patients with 17 lesions, with a matched comparison of a V6 surface lead and the RV to left upper chest, "intracardiac" lead. The intracardiac lead was sensitive in detecting myocardial injury current/ischemia. There was a 36.4+/-5.6% ST segment shift, relative to the amplitude of the QRS complex, in the intracardiac lead at 2 min, compared with a 10.1+/-1.9% ST shift from a surface lead (P=.00011). The RV to left upper chest lead detected a >10% shift in ST segment within 2 min in 17 (100%) of 17 cases vs. 8 (47%) of 17 for a V6 surface lead. The intracardiac lead provided detection of ischemia in all three major epicardial coronary distributions. CONCLUSIONS: This study demonstrates the ability of an intracardiac (RV apex to left upper chest) lead to rapidly detect myocardial ischemia/injury during acute coronary occlusion in the setting of balloon angioplasty. The results of this study suggest that a simple implantable system resembling a ventricular pacemaker could be programmed to assist in the very early diagnosis of acute myocardial infarction. PMID- 16263352 TI - High rate of right ventricular infarction after ligation of mid left anterior descending artery in rats. AB - BACKGROUND: The left anterior descending artery (LAD) supplies the left ventricle in humans. LAD ligation has been commonly used in rats to induce left ventricular (LV) infarction for research purposes. However, the myocardial supply territories of LAD are not well established in rats. We measured the infarction zone in rats after ligation of the mid-LAD. METHODS: Twenty-four male Sprague-Dawley rats weighing 300-350 g were selected for LAD ligation for the induction of ischemic cardiomyopathy. The surgery was performed under full anesthesia. Left-sided thoracotomy was performed through cuts in the fifth and sixth ribs. Ligation of the LAD was performed 1 to 2 mm distal to a line between the left border of the pulmonary conus and the right border of the left atrial appendage. LAD was ligated after the first diagonal and septal branches. After 24 h, the hearts were removed and stained with Tetrazolium Tetrachloride (TTC) for the detection of infracted areas. RESULTS: Ligation of LAD induces 85% infarction of the right anterior free wall and anterior right ventricular septum and induces 100% infarction of the anterior free wall of the left ventricle and anterior septum. Infarction after LAD ligation extends all the way to the distal of the ligation site down to the apex of the heart. CONCLUSIONS: Mid-LAD ligation after the first septal and diagonal branches causes substantial right ventricular infarction in addition to LV infarct in rats. Therefore, the hemodynamic effect of right ventricle infarct should be considered in research involving LAD ligation in rats. PMID- 16263353 TI - Estimation of radiation dose received during treatment of in-stent restenosis using ionizing radiation. AB - BACKGROUND: Angioplasty is a widely accepted procedure for the treatment of coronary artery disease. However, restenosis of the treated vessel occurs in 30% of patients within 6 months. Intravascular brachytherapy (IVB) is used to inhibit the formation of new tissue growth at the vessel treatment site. IVB protocols using either gamma ray or beta particle emitting isotopes have been tested and approved. However, very little data are available on resultant whole-body dose and the potential for long-term radiation effects. METHODS: Using thermoluminescence dosimetry (TLD) devices, specifically lithium fluoride (LiF) doped with Mg, Cu, and P, the radiation dose on the surface of patients undergoing IVB was measured. The TLDs were positioned on the body to obtain a measure of the gamma dose at selected anatomic locations. Additionally, the skin dose from fluoroscopy was estimated. RESULTS: Measurements indicate that the average body dose on the skin surface from all TLDs, clinical requirements, and gamma source configurations varies from 0.95 mSv (95 mrem) at the head to 27.06 mSv (2706 mrem) at the sternal notch. For beta sources, the dose varied from 0.11 mSv (11.4 mrem) at the head to 0.49 mSv (49.5 mrem) at the sternal notch. The fluoroscopy contribution of dose to the body dose (15-min exposure time) was 0.10 mSv (10 mrem) at the head and 2.57 mSv (257 mrem) to the sternal notch. CONCLUSIONS: The results suggest that surface skin exposures from gamma sources used in IVB pose acceptable risks considering the medical benefits of the procedures. PMID- 16263354 TI - Diagnostic modalities for the evaluation of anomalous left main coronary arteries. AB - Anomalous coronary arteries are rare and usually identified as an incidental finding during cardiac catheterization. The particular difficulty with cardiac catheterization techniques is not necessarily the presence of the anomalous coronary artery, but its anatomic course. Oftentimes, surgical intervention is necessary once these anomalies are discovered. The identification and anatomic characterization of anomalous coronary arteries has been significantly advanced with the use of current diagnostic noninvasive imaging modalities. We present 3 cases of an anomalous left main coronary artery that arises from the right sinus of Valsalva. Noninvasive imaging methods provided a clear anatomic course of the anomalous vessel. PMID- 16263355 TI - Drug-eluting stents for the treatment of in-stent restenosis: a clinical review. AB - Treatment of in-stent restenosis (ISR) remains problematic despite the widespread application of drug-eluting stents (DES). Challenging lesion cohorts such as diffuse ISR and restenosis after failed intracoronary radiation therapy (IRT) maybe best treated with DES. The overall benefit of DES appears inferior to their utility in treating de novo coronary lesions. Randomised trials comparing DES and IRT will soon be available to determine the optimal therapy for ISR. The challenge to treat ISR in the DES era is the next frontier of interventional cardiology. PMID- 16263356 TI - MRI evaluation of a coronary artery perforation. PMID- 16263358 TI - Evidence against the widespread use of angiography of noncoronary arteries during coronary artery angiography and cardiac catheterization. AB - PURPOSE: Although the most widely used screening techniques for extracardiac atherosclerotic distributions are noninvasive, in patients undergoing coronary arteriography, the question as to whether angiography of extracardiac arteries at the time of cardiac catheterization is really effective or if it should be considered malpractice is still open. We sought to determine the safety and real usefulness of angiography of extracardiac arteries performed by trained invasive cardiologists during cardiac catheterization. METHODS: Medical records of all patients undergoing combined coronary and noncoronary angiography between May 1998 and December 2002 were retrospectively reviewed. Moderate to severe arterial stenosis (>50% stenosis), vessel occlusion, aneurysmal, or severe ectasia were noted as significant angiographic findings. RESULTS: Two hundred and seventy patients (165 males, mean age = 67.7+/-9.2 years, mean serum creatinine = 1.1+/ 0.8 mg/dl) underwent combined cardiac catheterization and angiography of extracardiac arteries following specific indications. Significant findings were reported in 66 (24.4%) patients. Logistic regression analyses revealed three vessel coronary artery disease [CAD; odds ratio (OR)=9.917; 95% confidence interval (CI) = 2.2 to 43.8; P = .002) and hypercholesterolemia (OR = 2.851; 95% CI=1.03 to 7.9; P = .044) to be independent predictors of extracardiac atherosclerotic involvement. Complications rate was negligible. The detection of significant angiographic findings led to endovascular treatment in 37.8% and surgical vascular repair in 33.3% of cases. CONCLUSIONS: Angiography of extracardiac arteries at the time of coronary angiography appears justified only in patients with specific indications, multivessel CAD, and hypercholesterolemia. PMID- 16263360 TI - Segmental vessel wall shear stress and neointimal formation after sirolimus eluting stent implantation: physiological insights in a porcine coronary model. AB - BACKGROUND: Low vessel-wall shear stress promotes atherosclerosis and restenosis. We conducted serial analysis of vessel-wall shear stress following placement of metal and sirolimus (SRL) stents to determine the relationship between shear stress and neointima. METHODS: Serial quantitative coronary angiography, intracoronary ultrasound (IVUS), and Doppler flow analysis were performed at baseline, immediately poststent, and at 30 and 90 days on 16 stents (metal, n = 8; SRL, n = 8) implanted in the coronary arteries of eight miniswine. Segmental vessel-wall shear stress (dyn/cm2) was calculated at 10 sections within the stent and normalized to the average proximal and distal reference vessel shear stress using IVUS and hyperemic average peak flow velocity. At 90 days, histological analysis was completed to determine vessel-wall morphometry on corresponding sections from each stent. RESULTS: Stent placement resulted in a similar degree of in-stent stenosis (-5% to 25%) and immediate post-in-stent shear stress. At 30 days, the IVUS neointimal cross-sectional area and percentage of area stenosis were significantly less in SRL (1.2+/-0.8 mm2; 12.7+/-8.5%) versus metal stents (2.3+/-0.4 mm2; 28.2+/-3.4%, P < .003). In-stent normalized shear stress was less for SRL (0.93+/-0.07) versus metal (1.07+/-0.08, P = .002) stents. At 90 days, the mean neointimal area was similar for the SRL (2.50+/-0.47 mm2) and metal stents (2.72+/-1.15 mm2). Linear regression documented a negative correlation between poststent shear stress and neointima for metal stents (r = .61, P < .0001). In the SRL stents, however, the post-in-stent shear stress had a positive correlation with neointima (r = .40, P = .0002). CONCLUSIONS: The placement of oversized stents causes alteration of segmental vessel-wall shear stress, which appears to be an important physiological stimulus for neointimal formation, and may influence the pharmacodynamics of SRL-eluting stent in the porcine coronary model. PMID- 16263359 TI - Intracoronary radiation therapy using a novel beta emitter for in-stent restenosis Tungsten WRIST. AB - BACKGROUND: Intracoronary beta-radiation therapy reduces in-stent restenosis (ISR). We aimed to determine the safety and feasibility of intracoronary radiation therapy (IRT) utilizing tungsten (188W), a beta emitter. METHODS: A total of 30 patients with angiographic evidence of ISR in a previously treated native coronary artery underwent percutaneous coronary intervention (PCI; balloon angioplasty, ablation by atherectomy, or laser angioplasty). After the intervention, a noncentered delivery catheter with a side guide 0.014-in. wire carrying a tungsten (188W) coil, with an active length of 33 mm, was inserted. Patients were randomized to a radiation dose of 18, 22, or 25 Gy at 2 mm from the center of the source. Aspirin and Plavix, at 300 mg loading dose, were administered prior to intervention. Plavix 75 mg/day was prescribed for 6 months after the procedure. RESULTS: At 6 months follow-up, the overall binary angiographic restenosis rate was 18.8%. Target vessel revascularization (TVR) was 23% and target lesion revascularization related major adverse cardiac events (TLR MACE) was 13.3%, without any intergroup differences. A comparison with the original Washington Radiation for In-stent restenosis Trial (WRIST) radiation cohort utilizing an 192Iridium source (prescription dose 15 Gy at 2 mm from the source) showed similar TVR and TLR-MACE rates of 30% and 18%, respectively. The TVR and TLR-MACE rates in the WRIST placebo cohort were 70% and 66%, respectively. CONCLUSIONS: Vascular brachytherapy with tungsten (188W) is feasible and safe. The 6-month clinical outcomes are similar to the original WRIST radiation group. PMID- 16263361 TI - Human-grade purified collagenase for the treatment of experimental arterial chronic total occlusion. AB - PURPOSE: Chronic total occlusions (CTO) remain a major limitation of percutaneous interventions. Procedural failure is usually due to the inability to cross the lesion with a guide wire. We have previously shown that local administration of a laboratory-grade collagenase followed by a 72-h waiting period may facilitate guide-wire crossing. The aim of the present study was to evaluate the efficacy and toxicity of a human-grade purified collagenase, suitable for clinical use, in facilitating guide-wire crossing in a rabbit model of femoral artery CTO. METHODS AND RESULTS: A chronic total arterial occlusion was constructed in femoral arteries of New Zealand white rabbits. The local administration of purified collagenase solution (150 microg) via an over-the-wire balloon system was performed in 10 CTO. Guide-wire crossing was attempted after 24 h and was successful in all cases. Different doses (50-500 microg) were administered to an additional 17 rabbits to assess collagenase effects. Local subcutaneous bruising was observed at higher doses. Histological evaluation showed no damage to the arterial wall structure. Arterial extracts from collagenase-treated arteries showed increased MMP-2 and MMP-9 activities and higher levels of local MMP-1 and degraded collagen. CONCLUSIONS: Local administration of a human-grade purified collagenase degrades collagen in CTO and is highly effective for the facilitation of guide-wire crossing in CTO. PMID- 16263362 TI - Cardiovascular and hemodynamic effect of polyethylene glycol in rats. AB - BACKGROUND: Polyethylene Glycol (PEG) is a solvent and used in a wide range of biomedical applications. Many fatty-acid-based molecules cannot be administered without a solvent in vivo. PEG can be used to dissolve compounds to make them water soluble. However, the effect of PEG on the cardiovascular system has not been studied. In this study, we evaluated the effect of PEG on the cardiovascular system in rat models. METHODS: Twenty male Sprague-Dawley rats weighing 250-300 g were used in this study. The control group (10 rats) were injected intraperitoneally with 0.5 ml of 5% D/W in normal saline and the second group (10 rats) with PEG 400, 2 ml/kg ip, twice a day for 1 week. After 4 weeks, the rats underwent general anesthesia and a 1.4 French ultra miniature pressure volume catheter (Millar catheter) was placed in the left ventricle via the right carotid artery to measure comprehensive hemodynamic data. The data were analyzed with PVAN pressure-volume analysis software. RESULTS: All the systolic and diastolic parameters were similar in both groups except for the effective arterial elastance (Ea), which was decreased in the PEG group. There were no significant differences in maximum (dp/dt(max)) and minimum (dp/dt(min)) development of pressure stroke work, cardiac output, ejection fraction, end systolic volume (Ves), and end diastolic volume. CONCLUSIONS: We have demonstrated that PEG, as a solvent, decreases Ea in rats in comparison to a placebo. Therefore, PEG as a solvent should be used cautiously in the cardiovascular research. PMID- 16263363 TI - Percutaneous treatment of a giant coronary aneurysm using a Symbiot stent. AB - Percutaneous treatment of coronary aneurysms has been proven to be a safe and effective alternative to surgery. We report the successful treatment of a giant proximal left circumflex artery aneurysm with distal occlusion in a 76-year-old man using the new generation of covered stent, Symbiot, a polytetrafluoroethylene covered self-expandable stent (Boston Scientific, Natick, MA). PMID- 16263365 TI - Contrast-induced nephropathy: a review. AB - Contrast-induced nephropathy (CIN) is one of the leading causes of renal impairment in the United States and the third cause of hospital-acquired renal failure. Reduction in the incidence of CIN can lead to a decrease in the morbidity, mortality, and length of hospital stay. Although prophylactic hydration has been promising in decreasing the occurrence of CIN, other efforts such as diuretics, calcium channel blockers, theophylline, aminophylline, atrial natriuretic peptide, dopamine, and fenoldopam have been disappointing. The preventive effect of N-acetylcysteine on CIN has not been consistent in the literature. In a recent clinical trial, bicarbonate infusion was more effective than hydration in the prevention of CIN. Mechanical devices are in development to perfuse renal arteries with protective drugs during contrast exposure or for removal of contrast from coronary sinus during coronary angiography. In this article, we have reviewed available data in regards to CIN. PMID- 16263364 TI - Beta radiation in the treatment of in-stent restenosis of an in situ saphenous vein bypass graft A case report. AB - We describe a case of instent restenosis in a femoral-distal saphenous vein bypass graft successfully treated with brachytherapy. A 45-year-old insulin requiring diabetic woman underwent an in-situ femoral-anterior tibial bypass graft for a non-healing ischemic ulcer. Despite a technically successful percutaneous transluminal angioplasty and endovascular stenting of a retained valve within the threatened graft, the wound failed to heal. At the 1-month follow-up, instent restenosis was documented and successful cutting balloon angioplasty, complemented by adjunctive beta-irradiation was successfully performed. Clinical and hemodynamic success was achieved, with prompt ulcer healing and intermediate-term graft patency maintained on surveillance duplex ultrasound follow-up. We review the literature on radiation therapy in the management of peripheral arterial disease and discuss therapeutic options in the management of restenosis. PMID- 16263367 TI - The kidney, cancer, and the eye: current concepts. AB - Herein, we review the associations between the kidney, renal cancers, and the eye. Renal cancers have been reported to metastasize to the eye and the orbit. As these tumors can be confused with other amelanotic or vascular tumors, a high index of suspicion is required for early detection and management of the primary tumor. We discuss the physiology of metastases, clinical features and management of metastatic disease. A variety of ocular anomalies have been associated with renal disease. Wilms tumor, a renal tumor of childhood, can present with aniridia, which may be the first clue leading to the diagnosis of the primary tumor. Paraneoplastic syndromes are common manifestations of renal cancers and can present as retinopathies and neuro-ophthalmic disorders. Multiple cancer syndromes involve both the eye and the kidney. For example, the diagnosis of von Hippel retinal tumors can lead to a systemic evaluation and discovery of associated visceral tumors. The prognosis, screening, and counseling of such patients is discussed. Newer systemic treatments available for renal tumors, such as interferon alfa, may lead to ocular side effects including retinopathy. These patients require periodic ophthalmic examinations. This review demonstrates the essential role of the ophthalmologist, for early diagnosis and treatment that can help reduce the morbidity and mortality associated with kidney tumors and renal associated disease. PMID- 16263368 TI - Birdshot chorioretinopathy. AB - Birdshot chorioretinopathy is a well-known, yet poorly understood, form of posterior uveitis, characterized by multiple, distinctive, hypopigmented choroidal lesions, and strongly associated with human leukocyte antigen (HLA) A29. We reviewed all English language publications regarding birdshot chorioretinopathy and performed analyses of combined patient data taken from these articles. The mean age at presentation was 53 years, with a slight female predominance (54.1%). At least 95.7% of reported patients have been HLA-A29 positive. Blurring of vision and floaters are the most prevalent presenting complaints, even in patients with visual acuity of 20/20 or better in both eyes. Birdshot chorioretinopathy is a slowly progressive disease with profound dysfunction of vision that may not be reflected in Snellen visual acuity. Two or more lines of Snellen visual acuity were lost in approximately 20% of eyes over a median follow-up of 3.5 years; macular edema was the most common cause of reduced visual acuity. Overall, patients had a slow decline in visual acuity, despite the fact that nearly all were treated with anti-inflammatory therapies. Final visual acuity in the better eye was 20/40 or better in 75.1% of patients and 20/200 or worse in 9.8% of patients. Oral corticosteroids and cyclosporine were the most commonly used medications. Using a regression model, patients in the literature that have been treated with cyclosporine alone had better final visual acuity than patients treated with oral corticosteroids alone. Further study is needed to determine the optimal methods for treating and monitoring patients with birdshot chorioretinopathy. PMID- 16263369 TI - Using the Journal Club to teach and assess competence in practice-based learning and improvement: a literature review and recommendation for implementation. AB - The traditional journal club has historically been used to teach residents about critically reading and reviewing the literature in order to improve patient care. The Accreditation Council for Graduate Medical Education competencies mandate requires that ophthalmology residency programs both teach and assess practice based learning and improvement. A systematically conducted review of the literature regarding the use of the journal club in resident medical education was performed to define specific recommendations for implementation of a journal club tool. Selected best practices for a successful journal club were gleaned from the existing medical literature. These include the following: 1) the use of a structured review checklist, 2) explicit written learning objectives, and 3) a formalized meeting structure and process. The journal club might prove to be an excellent tool for the assessment of competencies like practice-based learning which may be difficult to assess by other means. Future study is necessary to determine if journal club can improve educational outcomes and promote lifelong competence in practice-based learning. PMID- 16263370 TI - Phakic intraocular lenses. AB - An analytical review of the data available in the field of phakic intraocular lens implantation was conducted. Particular attention was paid to the more critical issues of intraocular lens sizing and safety guidelines. A comprehensive, competitive analysis of different implantation sites, intraocular lens model designs, and safety guidelines has been included. Specialized biometry techniques, such as very high frequency ultrasound and Scheimpflug imaging, have been reviewed, and a critical review of commercial claims regarding intraocular lens technologies has been included. Clinical studies of phakic intraocular lenses demonstrate increasing promise for the correction of refractive errors not amenable to mainstream excimer laser refractive surgery. The main issues currently revolve around adequate lens design (VHF ultrasound study suggests that custom-design and sizing may be the most effective and safest approach for every phakic IOL model), because these devices will be required to remain physiologically inert and anatomically compatible with internal ocular structures and relations for several decades. The possibility of safe removing or exchanging the IOL should remain a feasible option over time. It is of utmost importance that we continue to critically evaluate current encouraging short-term outcomes, which are being extrapolated to the longer term by ongoing high resolution imaging and monitoring of the anatomical and functional relations of implanted phakic IOLs. PMID- 16263371 TI - Her vision was tied down. AB - A 50-year-old female presented with progressive painless vision loss in the left eye and was subsequently diagnosed to have a pituitary macroadenoma, consistent with a prolactinoma, which was compressing the chiasm primarily on the left and involving the left cavernous sinus. She was treated with oral bromocriptine, resulting in marked shrinkage of the tumor and significant visual field recovery. Subsequently, she again began noting progressive vision loss in both eyes and was initially thought to have pituitary tumor regrowth. Repeat brain MRI, however, showed chiasmal prolapse with inferior tethering into an empty sella. Surgical fat padding to reposition the chiasm resulted in visual improvement with a different visual field pattern in the left eye. The clinical manifestations and etiopathogenesis of the empty sella syndrome are discussed. PMID- 16263372 TI - Isolated cranial nerve III palsy as the presenting manifestation of HIV-related large B-cell lymphoma: clinical, radiological and postmortem observations: report of a case and review of the literature. AB - Rarely can a neurologically isolated cranial nerve III palsy be the presenting manifestation of central nervous system lymphoma. We detail the clinical, radiological, and pathological features of a previously healthy 45-year-old man presenting with an isolated, pupil-involving, right cranial nerve III palsy due to human immunodefiency virus (HIV) related non-Hodgkin lymphoma. Magnetic resonance imaging demonstrated bilateral peripheral cranial nerve III enhancement with no brain parenchymal or leptomeningeal abnormalities. Cerebrospinal fluid analysis revealed a monocytic pleocytosis with an elevated protein concentration and depressed glucose level. Morphologic and flow cytometric analysis of the cerebrospinal fluid was compatible with a large B-cell lymphoma. Serologic tests for HIV were positive. Postmortem examination of the brain revealed malignant lymphomatous cell infiltration of both cranial nerve III, diffuse leptomeningeal disease and focal superficial subependymal and subpial invasion. Based on our review of the literature, we were able to find only 10 detailed cases of cranial nerve III palsy as the presenting manifestation of central nervous system lymphoma. Furthermore, none of the previously reported cases correlated the magnetic resonance imaging findings with the gross and histopathologic observations. PMID- 16263376 TI - Toxicity of surfactants to green microalgae Pseudokirchneriella subcapitata and Scenedesmus subspicatus and to marine diatoms Phaeodactylum tricornutum and Skeletonema costatum. AB - Ecotoxicity of different commercial surfactants (six anionic, two amphoteric and one nonionic), essential constituents of cleansing hair products (shampoos), as well as ecotoxicity of eight shampoos containing different combinations of these surfactants, were tested in order to evaluate their possible toxic effects on microalgae. Specific objective of this research was to compare the sensitivity of selected freshwater and marine microalgae to these widely used surfactants and well-known pollutants in surface waters. Internationally validated methods (ISO standards) for the determination of toxic effects on the growth of planktonic freshwater green algae Pseudokirchneriella subcapitata and Scenedesmus subspicatus and marine diatoms Skeletonema costatum and Phaeodactylum tricornutum, were used. The obtained results showed that the concentrations of tested surfactants and shampoos, which resulted in 50% growth reduction of planktonic freshwater green algae, when compared to the controls without test substances (EC50), were in the range from 0.32 to 4.4 mg l(-1) for surfactants and from 2.1 to 8.5 mg l(-1) for shampoos expressed as active substance. Marine diatoms were significantly more sensitive to the tested surfactants than freshwater green algae (EC50 0.14-1.7 mg l(-1) for surfactants and 0.35-1.25 mg l(-1) for shampoos). According to the classification on the basis of environmental effects, the obtained results suggested that all tested surfactants can be classified as having toxic effects on freshwater green alga Pseudokirchneriella subcapitata. Some of them indicated that they have a very toxic effect on Scenedesmus subspicatus and marine diatoms Skeletonema costatum and Phaeodactylum tricornutum. PMID- 16263374 TI - Considerations of glaucoma patients undergoing corneal refractive surgery. PMID- 16263377 TI - Distribution and toxicity of mercury in rats after oral administration of mercury contaminated whale red meat marketed for human consumption. AB - Toothed-whales and dolphins have been hunted for human consumption in Japan, and their muscles (red meats) are highly contaminated with mercury (Hg). We investigated the distribution and toxicity of Hg in rats after oral administration of Hg-contaminated whale red meat marketed for human consumption in Japan. Rats were orally administered the red meat homogenate for seven consecutive days (0.5 g red meat/kg-bw/day). The red meat administered to rats contained 81microg/g of total mercury (T-Hg) and 13.4 microg/g of methyl mercury (M-Hg). This dose corresponds to the human consumption of 210 g red meat/60 kg bw/week, exceeding by about 29 times the provisional tolerable weekly intake of M Hg at 1.6 microg/kg-bw/week set by JECFA [JECFA, 2003. Joint FAO/WHO expert committee on food additives. 61st meeting, Rome]. Twenty-four hours after the last administration, the distribution of T-Hg in rat organs and biochemical parameters in serum were analyzed. The administration of red meat significantly elevated T-Hg concentrations in the liver, kidney, erythrocytes, cerebral cortex and medulla oblongata from the control levels but did not elevate the T-Hg concentration in serum, showing the typical distribution pattern of M-Hg, not of inorganic Hg. The administration slightly but significantly increased GTP activity and P concentration and decreased BUN concentration in serum, although no abnormalities were observed in rat body weight gain and movement during the 7 days. The occasional consumption of red meat from small cetaceans, therefore, could pose a health problem for not only pregnant women but also for the general population. PMID- 16263378 TI - Screening of POP pollution by AChE and EROD activities in Zebra mussels from the Italian Great Lakes. AB - The increase of ethoxyresorufin dealkylation (EROD) and the inhibition of acetylcholinesterase (AChE) as biomarkers have been commonly used in vertebrates for the persistent organic pollutants (POPs) biomonitoring of aquatic environments, but very few studies have been performed for invertebrates. Previous researches demonstrated the interference due to some chemicals on EROD and AChE activities of the freshwater bivalve Zebra mussel (Dreissena polymorpha) in laboratory and field studies, showing its possible use for the screening of POP effects. We investigated the contamination of the Italian sub-alpine great lakes (Maggiore, Lugano, Como, Iseo, Garda) by the biomarker approach on Zebra mussel specimens collected at 17 sampling sites with different morphometric characteristics and anthropization levels. Results showed a homogeneous contamination of AChE inhibitors in Lake Garda, Maggiore, Como and Iseo with values ranging from 0.5 to 3 nmol/min/mg proteins and with an average inhibition of about 66% to controls. The planar compounds pollution, able to activate the EROD activity, seems higher in some sampling stations of Lake Garda, Como and Iseo (2-4 pmol/min/mg proteins) than that measured in Lake Lugano (1.5-3 pmol/min/mg proteins). On the contrary, the enzyme activity in Lake Maggiore showed an interesting opposite effect of AhR-binding compounds and trace metals. Finally, the possible use of Zebra mussel specimens maintained at laboratory conditions as controls against the selection of the less polluted sampling site is discussed. PMID- 16263379 TI - Speciation of inorganic arsenic in alimentary and environmental aqueous samples by using derivative anodic stripping chronopotentiometry (dASCP). AB - The purpose of this research was to develop a sensitive and accurate chronopotentiometric method at a gold film electrode, to determine trace and ultra trace levels of As(III) and As(V) in alimentary and environmental water systems. As(III) was directly determined in the aqueous matrix at a deposition potential of -300 mV for 180 s and at a constant anodic current of 2.5 microA, without any sample pre-treatment; moreover the chronopotentiometric method did not require a time-consuming de-oxygenation step prior to the analysis. A 3M HCl solution was chosen as the best stripping medium. The direct analysis of As(V) required the application of a high negative over-potential and, thus, measurements were characterized by poor reproducibility; therefore As(V) was determined after reduction to As(III) with KI in a strong hydrochloric acid solution. Under the optimised electrochemical conditions, detection limits of 0.08 microg As(III) l(-1) were achieved and no significant interferences from Cd, Cu, Pb, Zn and organic substances were observed. As(V) was the most abundant species in all the studied environmental and alimentary aqueous matrices. Amongst the beverages, tea and coffee presented the As(V) highest concentration ranges (934-1740 microg l(-1) and 850-1290 microg l(-1), respectively) while bottled mineral water the lowest (<1.61 microg l(-1)); whereas As(III) levels lower than 5.0 microg l(-1) were detected only in wine samples. PMID- 16263380 TI - Effect of different soil types on the enchytraeids Enchytraeus albidus and Enchytraeus luxuriosus using the herbicide Phenmedipham. AB - Soil ecotoxicology studies are usually performed in standard soils, such as the OECD (Organization for Economic Cooperation and Development) artificial soil or the LUFA St. 2.2, a natural soil. When assessing the toxic effects in the environment, soil properties are often different from those in standard soils, which might lead to a different exposure situation for the test species and, therefore, to a different evaluation of the risk of the test substance. Selected to cover a broad range of properties and based on the Euro-Soils concept, 18 different soils were studied regarding their suitability to two test species: Enchytraeus albidus and Enchytraeus luxuriosus (Enchytraeidae). In reproduction tests, the test species reacted differently to the tested soils, but both enchytraeids did not survive in acid soils (i.e. pH600 mg/l, respectively. Separate 30-d exposure experiments yielded no observed and lowest observed effect concentrations, respectively, of 0.25 and 0.5mg/l for MagnaFloc 368, and 75 and 150 mg/l for MagnaFloc 156. In both cases, behavioural responses, especially startle response, were the most sensitive test endpoints. Histopathological assessment revealed that gill pathology appeared within a few days of exposure to both polymers, apparently as a result of localized hypoxia. Acute (4 d) effects included cloudy swelling of epithelial cells, increased gill vascularization, and thickening and shortening of the gill lamella. Chronic (30 d) polymer exposure produced only slightly greater pathological effects than acute exposure, with comparable responses observed only at >1.0mg/l MagnaFloc 368 and 150 mg/l MagnaFloc 156, suggesting that the fish displayed some level of both behavioural and physiological adaptation to the respiratory stress imposed by the two polymers. PMID- 16263383 TI - Mutagenicity of anaerobic fenitrothion metabolites after aerobic biodegradation. AB - Previous studies have revealed that the mutagenicity of fenitrothion increases during anaerobic biodegradation, suggesting that this insecticide's mutagenicity could effectively increase after it pollutes anaerobic environments such as lake sediments. To investigate possible changes to the mutagenicity of fenitrothion under aerobic conditions after it had already been increased by anaerobic biodegradation, batch incubation cultures were maintained under aerobic conditions. The mutagenicity, which had increased during anaerobic biodegradation, decreased under aerobic conditions with aerobic or facultative bacteria, but did not disappear completely in 22 days. In contrast, it did not change under aerobic conditions without bacteria or under continued anaerobic conditions. These observations suggest that the mutagenicity of anaerobically metabolized fenitrothion would not necessarily decrease after it arrives in an aerobic environment: this would depend on the presence of suitable bacteria. Therefore, fenitrothion-derived mutagenic compounds may pollute the water environment, including our drinking water sources, after accidental pollution of aerobic waters. Although amino-fenitrothion generated during anaerobic biodegradation of fenitrothion was the principal mutagen, non-trivial contributions of other, unidentified metabolites to the mutagenicity were also observed. PMID- 16263384 TI - Ecological risk assessment for aquatic species exposed to contaminants in Keelung River, Taiwan. AB - An ecological risk assessment was conducted for Keelung River in northern Taiwan. The objective of this study was to assess the risk to fish, aquatic insects, and benthic macroinvertebrates associated with chemical-of-potential-concern (COPC) in the river and to rank ecological risk for these chemicals. The protection of at least 95% of the species 90% of the time from acute and chronic COPC exposures was the defined assessment endpoint. Nine inorganic and organic contaminants were selected to evaluate the impact to aquatic community in the Keelung River. The quotient method served as screen level estimation of risk. The Aquatic Ecological Risk Assessment model was used to analyze exposure and ecological effects and to estimate community level risk. The logarithmic regression model between probability and lethal concentration was established. The combined risks of multiple chemicals were evaluated under assumption of additive risk. The results indicated that zinc and copper pose higher risk among metals. Ammonia, copper, and zinc posed virtually all of the risk, while organic COPCs posed a negligible risk. Potential ecological risk from ammonia exposure was greatest. The probability of more than 5% of the species being affected by acute or chronic toxicity of COPCs is about 100%. In average (50% of the time), 99% of the species would be affected by acute toxicity of COPCs, and about all the species would be affected by chronic toxicity of COPCs. Uncertainties in this assessment were associated with variability in ecosystem stressors, exposure data, ecological effect data, and risk characterization. PMID- 16263385 TI - Inter- and intra-species variation in acute zinc tolerance of field-collected cladoceran populations. AB - Acute zinc toxicity was assessed for 10 freshwater cladoceran species collected in six different ecosystems across Europe and for two standard laboratory-reared species (Daphnia magna and Ceriodaphnia dubia). The collected organisms belonged to five different genera: Daphnia (subgenus Daphnia and Ctenodaphnia), Ceriodaphnia, Simocephalus, Acroperus and Chydorus. The 48-h EC50 of the field collected organisms tested in standard laboratory water ranged from 375+/-141 to 4314+/-1513 microg Znl(-1). The laboratory clone of D. magna was less sensitive than the majority of the field-collected species, while our laboratory Ceriodaphnia dubia was the second most sensitive. Considerable inter-species variation was found within the genus of Ceriodaphnia (factor 6) and within the genus Daphnia (factor 8). Among the different (sub)genera tested, Chydorus and Ctenodaphnia were significantly more tolerant than the others (up to a factor 3 difference). A significant positive relationship (r2=0.67, p<0.05) between the mean cladoceran 48-h EC50 and the ambient zinc concentration of the different aquatic systems was demonstrated, suggesting a role of acclimation and/or adaptation. No significant correlation between the acute zinc tolerance and the length of the organisms was found. PMID- 16263386 TI - Production of male neonates in Daphnia magna (Cladocera, Crustacea) exposed to juvenile hormones and their analogs. AB - We exposed the water flea Daphnia magna (Cladocera, Crustacea) to either juvenile hormone I (JH I), juvenile hormone II (JH II), or the juvenile hormone-mimicking insecticides kinoprene, hydroprene, epofenonane, or fenoxycarb. By 21-day reproduction tests, we investigated the effects on the number of neonates born per female and the offspring sex ratio. All six chemicals induced D. magna to produce male neonates; the male sex ratio of the offspring increased as the chemical concentration increased. EC50 values for production of male neonates were estimated as 400 (JH I), 410 (JH II), 190 (kinoprene), 2.9 (hydroprene), 64 (epofenonane), and 0.92 (fenoxycarb) microg/l. The number of neonates produced was reduced with all chemicals at the concentrations investigated. At the EC50 for male production, five of the six chemicals reduced the reproductive rate to less than 50%; the exception was epofenonane, which caused only a slight reduction in reproductive rate. These results were similar to those obtained for five juvenoids studied previously, one of which was studied here again. There are now 10 chemical substances--all juvenile hormones or their analogs-that are known to induce D. magna to produce male neonates. This suggests that juvenile hormone is involved in initiating male production followed by sexual reproduction in D. magna, and probably in most cladocerans that exhibit cyclic parthenogenesis. PMID- 16263387 TI - Interaction of polycyclic aromatic hydrocarbons and heavy metals on soil enzyme. AB - Actions and interactions of heavy metals (cadmium, zinc and plumbum) and polycyclic aromatic hydrocarbons (PAHs) [phenanthrene, fluoranthene, benzo(a)pyrene] on the soil urease and dehydrogenase activity were studied after 49 days exposure. The experimental approach was based on the uniform design which can cut the experiment time and improve the efficiency of experiments. Data treatment was essentially based on the multiple regression technique. The results showed that the action and interaction between heavy metals and PAHs were strongly dependent on the time of pollution. The dehydrogenase exhibits more sensitive to the combined pollution than urease. The negative interaction between Zn and Cd to hydrogenase activity and the combined stimulatory activity of Phenanthrene and Benzo(a)pyrene (or fluoranthene) to soil enzyme were observed. The interactions between Zn (Cd) and phenanthrene towards urease (dehydrogenase) were positive, and the interaction between Zn and benzo(a)pyrene to urease activity was negative. This study corresponds to exploratory phase in order to reveal interaction effects of heavy metals and PAHs on the soil enzyme and then to set up more in-depth analysis to increase progressively the understanding of the ecotoxicological mechanisms involved. PMID- 16263388 TI - Environmental signals: synthetic humic substances act as xeno-estrogen and affect the thyroid system of Xenopus laevis. AB - According to outdated paradigms humic substances (HS) are considered to be refractory or inert that do not directly interact with aquatic organisms. However, they are taken up and induce biotransformation activities and may act as hormone-like substances. In the present study, we tested whether HS can interfere with endocrine regulation in the amphibian Xenopus laevis. In order to exclude contamination with phyto-hormones, which may occur in environmental isolates, the artificial HS1500 was applied. The in vivo results showed that HS1500 causes significant estrogenic effects on X. laevis during its larval development and results of semi-quantitative RT-PCR revealed a marked increase of the estrogenic biomarker estrogen receptor mRNA (ER-mRNA). Furthermore, preliminary RT-PCR results showed that the thyroid-stimulating hormone (TSHbeta-mRNA) is enhanced after exposure to HS1500, indicating a weak adverse effect on T3/T4 availability. Hence, HS may have estrogenic and anti-thyroidal effects on aquatic animals, and therefore may influence the structure of aquatic communities and they may be considered environmental signaling chemicals. PMID- 16263390 TI - Effects of heavy metals on sea urchin embryo development. Part 2. Interactive toxic effects of heavy metals in synthetic mine effluents. AB - Interactive toxic effects between heavy metals were investigated using a sea urchin (Anthocidaris crassispina) bioassay. An effluent from an abandoned mine showed significant inhibitory effects on embryo development as well as producing specific malformations. The effects on the embryos were reproduced by synthetic polluted seawater consisting of eight metals (manganese, lead, cadmium, nickel, zinc, chromium, iron, and copper) at the concentrations detected in the mine effluent. This indicated that the heavy metals were responsible for the effects observed. Five heavy metals were ranked in decreasing order of toxicity as follows: Cu>Zn>Pb>Fe>Mn. Among these, zinc and manganese could cause malformation of the embryos. From bioassay results using 27 combinations of heavy metals, 16 combinations including zinc could produce specific malformations, such as radialized, exo-gastrulal, and spaceship Apollo-like gastrulal embryos. Zinc was one of the elements responsible for causing malformations and its effects were intensified by the presence of the other metals, such as manganese, lead, iron, and copper. PMID- 16263389 TI - Resistance to metal contamination by historically-stressed populations of Ceriodaphnia pulchella: environmental influence versus genetic determination. AB - Field populations of daphnids historically-stressed by metal contamination may show increased resistance to those contaminants. This study was undertaken aiming to confirm/infirm three main hypotheses: (1) field populations living in historically-impacted environments are more tolerant to metal stress than populations from reference sites; (2) resistance differences are genetically determined, i.e., differences persist after controlling for environmental and maternal effects, by acclimating cloned lineages to similar conditions; and (3) resistance to stress in field populations living in historically-impacted environments is due to the disappearance of sensitive individuals rather than the appearance of highly resistant ones, i.e., the shift in the central tendency of resistance is linked to a decrease in the range of population resistance and not to an increased upper limit of the population resistance. Three populations of the cladoceran Ceriodaphnia pulchella Sars in Southern Portugal were sampled; one of which has been historically-stressed by acid mine drainage (AMD) from an abandoned cupric-pyrite mine and two from reference sites within the same watershed. To assess if resistance differences were genetically-determined, the three populations were acclimated for at least five generations under the same controlled conditions. Assays with AMD contaminated water samples were performed with both non-acclimated and acclimated individuals from all studied populations. Reproduction results in sub-lethal assays revealed significant differences between the reference and stressed populations. Significant differences in resistance to lethal levels of toxicity were observed for both non-acclimated and acclimated populations, individuals from population I being more resistant than those from reference populations. The existence of genetically-determined sensitivity differences was attested by the presence of significant differences in resistance to lethal levels of toxicity in acclimated individuals from reference and stressed populations. Results from cumulative mortality assays revealed that sensitive individuals were most probably present in the original population, but no conclusion could be draw about the presence of extreme resistant individuals in the historically-stressed population. Finally, it was shown that responses among populations converged from high to low levels of contamination. PMID- 16263393 TI - An overview of hematopoietic stem cell transplantation. AB - This article is a broad overview of hematopoietic stem cell transplantation, covering topics such as indications for transplantation, types of transplantation procedures, HLA matching of donors and recipients, procurement of stem cells from donors, and outcomes of transplantation. The major complications of transplantation, such as graft-versus-host disease, and the mechanisms of tissue injury after transplantation are discussed, as are the nature of immunologic impairment and reconstitution after transplantation. Finally future directions in stem cell transplantation are discussed. PMID- 16263394 TI - An overview of solid organ transplantation. AB - Once a medical curiosity, solid organ transplantation is now a commonplace occurrence, with more than 27,000 procedures performed in the United States in 2004 alone. This article offers an overview of the various solid organ transplant procedures to provide a context within which subsequent articles on pulmonary complications can be viewed. PMID- 16263395 TI - Pulmonary complications of transplantation: radiographic considerations. AB - The aim of this article is to clarify radiographic definitions associated with common parenchymal patterns encountered in the transplant population and to discuss the most common pathologic causes responsible for each pattern. The article also touches on radiographic findings signifying complications of other intrathoracic structures, including the airways, pleural space, and mediastinum. PMID- 16263396 TI - Acute lung injury after hematopoietic stem cell transplantation. AB - Hematopoietic stem cell transplantation (HSCT) has emerged as a common therapeutic option for a variety of life-threatening disorders, especially hematologic malignancies. Pulmonary complications are reported in 30% to 60% of all recipients and represent a major cause of mortality. A major proportion of these complications are the direct result of infection. This article addresses early, noninfectious causes of acute lung injury in the HSCT recipient. PMID- 16263397 TI - Chronic lung disease after hematopoietic stem cell transplantation. AB - Tens of thousands of patients undergo hematopoietic stem cell transplantation (HSCT) each year, mainly for hematologic disorders. In addition to the underlying diseases, the chemotherapy and radiation therapy that HSCT recipients receive can result in damage to multiple organ systems. Pulmonary complications develop in 30% to 60% of HSCT recipients. With the widespread use of prophylaxis for certain infections, the spectrum of pulmonary complications after HSCT has shifted from more infectious to noninfectious complications. This article reviews some of the noninfectious, chronic pulmonary complications. PMID- 16263398 TI - Hepatopulmonary syndrome and portopulmonary hypertension: implications for liver transplantation. AB - Hepatopulmonary syndrome (HPS) and portopulmonary hypertension (POPH) are uncommon pulmonary vascular consequences of advanced liver disease. HPS, characterized by arterial hypoxemia caused by pulmonary vascular dilatation, may resolve completely after liver transplantation. POPH, caused by vasoproliferation/constriction and obstruction of pulmonary arterial blood flow, is associated with higher risk for liver transplantation and increased post transplantation mortality. With or without transplantation, survival in patients who have these syndromes is associated with specific oxygenation and hemodynamic variables. PMID- 16263399 TI - Allograft rejection after lung transplantation. AB - Lung transplantation has become an accepted therapy for selected patients with advanced lung disease. One of the main limitations to successful lung transplantation is rejection of the transplanted organ. This article discusses the clinical presentation, treatment, and prevention of hyperacute, acute, and chronic rejection in the lung transplant recipient. PMID- 16263400 TI - Noninfectious pulmonary complications after lung transplantation. AB - This article reviews several important noninfectious pulmonary complications that threaten survival, pulmonary function, and quality of life after lung transplantation. Topics reviewed include primary graft dysfunction (PGD), native lung hyperinflation, anastomotic complications, phrenic nerve injury, pleural complications, lung cancer, pulmonary toxicity associated with immunosuppressive medications, and exercise limitation. PMID- 16263401 TI - Noninfectious pulmonary complications of liver, heart, and kidney transplantation. AB - Because of their chronically immunosuppressed status, solid organ transplant recipients are continually at risk for infectious pulmonary complications. In addition, however, a number of noninfectious pulmonary complications plague the transplant recipient. These complications arise because of numerous factors, including the underlying conditions that preceded transplantation, the transplant surgery itself, and toxicity of post-transplantation medications. This article focuses on noninfectious pulmonary complications in the three largest recipient populations: liver, kidney, and heart. PMID- 16263403 TI - Bacterial and mycobacterial pneumonia in transplant recipients. AB - Bacteria and myobacteria are important pulmonary pathogens in transplant recipients and are the focus of this article. Although considerable overlap exists, there are significant differences in the epidemiology and clinical presentation of these organisms in solid organ transplant (SOT) and hematopoietic stem cell transplant (HSCT) recipients. The first section of this article focuses on infections in SOT recipients (predominantly heart, liver, lung, and kidney transplant recipients), and the latter addresses these infections in HSCT recipients. PMID- 16263402 TI - Post-transplant lymphoproliferative disorder. AB - Post-transplant lymphoproliferative disorder (PTLD) is an increasingly recognized complication of solid organ and hematopoietic stem cell transplantation. PTLD represents a spectrum of polyclonal and monoclonal lymphoproliferation, generally of B cells. Prompt diagnosis is key and requires a high index of suspicion. An increasing variety of highly effective therapies, including immune modulation via reduction in immunosuppression, monoclonal antibodies, and cellular therapy, have dramatically improved the cure rates of this once devastating disease. PMID- 16263404 TI - Aspergillus pulmonary infections in transplant recipients. AB - Aspergillus infections are increasing in frequency in those undergoing solid organ and hematopoietic stem cell transplantation. The ongoing impact of Aspergillus infection on morbidity and mortality after transplantation makes this subject an area of intense clinical and research interest. This article discusses the evolving epidemiologic features of the infection and its management and diagnosis. PMID- 16263405 TI - Non-Aspergillus fungal pneumonia in transplant recipients. AB - Although species of Aspergillus and Candida account for most deeply invasive and life-threatening fungal infections, the past decades have seen a rise in the immunocompromised population. With this increase, additional fungi have emerged as important agents of morbidity and mortality. These opportunistic fungi are characterized by their ubiquitous presence in the environment, their ability to cause disease in immunosuppressed patients, and their diminished susceptibility to the currently available antifungal agents. Pneumonia, one aspect of a myriad of clinical manifestations caused by these fungal pathogens, is discussed in this article. PMID- 16263406 TI - Cytomegalovirus pneumonia in transplant recipients. AB - The incidence of cytomegalovirus CMV pneumonia has been reduced by routine antiviral prophylaxis in susceptible populations. Many of the complications of this infection are caused by indirect effects of the virus, including acute and chronic graft rejection, graft-versus-host disease, and superinfection by other viruses, bacteria and fungi. Distinction must be made between viral secretion and invasion. Invasive procedures are often required for the optimal management of such infections. The use of sensitive and quantitative assays have greatly improved the outcomes of CMV infection. PMID- 16263407 TI - Viral pneumonias other than cytomegalovirus in transplant recipients. AB - Community-acquired respiratory viruses (CARVs) are frequent causes of upper and lower respiratory tract infections in transplant recipients. In most series, respiratory syncytial virus and parainfluenza are the most common CARVs, followed by influenza and adenovirus. Significant morbidity and mortality are associated with these infections, particularly when they progress to pneumonia or when they are associated with bacterial or fungal coinfections. Outcomes are also poor with adenovirus, frequently reflecting disseminated infection. Efforts to prevent morbidity and mortality from CARV infection should focus on prevention, because treatment options are limited with inconclusive data to support their efficacy. PMID- 16263408 TI - Erythropoietin: the story of hypoxia and a finely regulated hematopoietic hormone. AB - The dramatic increase in knowledge during the last half century about the hormone erythropoietin is reviewed. The description of these events has been separated into two parts. The first part describes how the rapid changes in response to tissue oxygenation in the kidneys changes the rate of erythropoietin production. The second part describes how changes in erythropoietin concentrations act on erythroid progenitor cells, resulting in prompt changes in rates of erythrocyte production. Together these two aspects of erythropoietin biology provide an explanation for the tight physiological regulation of the numbers of circulating erythrocytes and, in a more general manner, provide a model for the control of the numbers of other specific blood cells by their respective hematopoietic growth factors. PMID- 16263409 TI - On the proper use of statistical design of experiments. PMID- 16263411 TI - Telomerase vaccination has no detectable effect on SCID-repopulating and colony forming activities in the bone marrow of cancer patients. AB - OBJECTIVES: The telomerase reverse transcriptase hTERT is a widely expressed tumor-associated antigen recognized by cytotoxic T lymphocytes (CTL). We have previously shown that vaccination of cancer patients against hTERT induces functional anti-tumor CTL in vivo, but it is not known whether hTERT vaccination harms normal cells expressing the enzyme, especially hematopoietic stem cells and progenitors. PATIENTS AND METHODS: We employed colony-forming cell (CFC) assays, long-term in vitro cultures, and nonobese diabetic/severe combined immunodeficient (NOD/SCID) repopulation studies to evaluate the effects of hTERT vaccination on hematopoietic progenitors and stem cells in cancer patients following treatment. RESULTS: Using bone marrow samples obtained from cancer patients before and after vaccination, we found that there was no significant decline in the frequency of granulocyte, macrophage or erythroid CFCs using CFC assays or long-term in vitro cultures. In NOD/SCID mice, human hematopoietic reconstitution was easily detected, without quantitative or qualitative differences between pre- and postvaccine samples. CONCLUSION: These findings suggest that induction of tumor-lytic hTERT-specific T cells in vivo by vaccination does not result in a detectable decline in hematopoietic potential despite the expression of hTERT and major histocompatibility complex class I in bone marrow progenitors and stem cells. Thus, even for self-antigens such as telomerase, tumor immunity does not necessarily involve autoimmunity in normal tissues that share the target. PMID- 16263412 TI - TGF-beta1 inhibits late-stage mast cell maturation. AB - OBJECTIVE: The objective of this study was to determine the effects of transforming growth factor (TGF)-beta1 on mast cell development. MATERIALS AND METHODS: Mast cells were cultured from mouse bone marrow in interleukin (IL)-3 + stem cell factor, in the presence or absence of TGF-beta1. We assessed mast cell development by measuring the expression of kit, T1/ST2, FcvarepsilonRI, and Fcgamma receptors. Cell morphology was determined by histochemical staining. Alterations in FcvarepsilonRI subunit expression were measured by Western blot analysis. Adoptive transfer of cultured mast cells into mast cell-deficient W/W(v) mice was used to determine if the in vivo environment could reverse the inhibitory effects of TGF-beta1. RESULTS: TGF-beta1 decreased FcvarepsilonRI, c kit, T1/ST2, and FcgammaR expression, and inhibited granule formation in developing mast cells. Accessory cells were not required for this inhibition. Smad3 deficiency did not alter the response of bone marrow cells to TGF-beta1. TGF-beta1 inhibited expression of the FcvarepsilonRI alpha subunit protein, without decreasing beta or gamma proteins. Mast cells derived in the presence of TGF-beta1 were functionally impaired, as IgE-mediated cytokine secretion was greatly reduced. The changes in granule formation and surface antigen expression were long-standing, as they were not reversed by transfer to W/W(v) mice. CONCLUSIONS: TGF-beta1 may contribute to mast cell homeostasis by inhibiting maturation from bone marrow precursors. The effects of TGF-beta1 result in greatly diminished expression of cell surface markers, reduced granulation, and lack of responsiveness to IgE-mediated activation. Thus TGF-beta1 can serve as a potent and multifunctional regulator of mast cell maturation. PMID- 16263413 TI - Ineffective erythropoiesis in mutant mice with deficient pyruvate kinase activity. AB - OBJECTIVE: A deficiency of pyruvate kinase (PK) is the most common cause of hereditary nonspherocytic anemia due to glycolytic enzyme defects. Red cells are poorly deformable due to adenosine triphosphate depletion in individuals with a PK deficiency and are destroyed in the microcirculation of the reticuloendothelial system, leading to extravascular hemolysis. The pathophysiology of PK deficiency has been widely studied in PK-deficient mice (PK 1(slc)). We examined the effects of a PK deficiency on erythroid progenitor maturation using these mice. MATERIALS AND METHODS: The appearance of apoptotic cells in spleen of PK-1(slc) mice was examined by terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) staining. We also assayed hematopoietic stem cell colony formation in vitro in the spleen of PK-1(slc) mice, to investigate erythropoiesis, and annexin V binding, as a measure of apoptotic cells in constitutive erythroid colonies, to evaluate the maturation of erythroid progenitors. RESULTS: The number of hematopoietic progenitors including colony-forming unit erythroids, burst-forming unit erythroids (BFU-E), colony forming unit granulocyte-macrophages, and multilineage colony-forming units in the spleens of PK-1(slc) was remarkably increased indicating hematopoiesis, and enhanced erythropoiesis in particular. TUNEL assays identified apoptotic cells in the splenic red pulp of the PK-1(slc) mice. Two-color flow cytometry detected apoptotic cells among anti-TER119-positive cells, suggesting that apoptotic cells were of erythroid lineage. Cells undergoing apoptosis were detected in cultures of BFU-E generated from bone marrow cells of PK-1(slc) mice. CONCLUSIONS: The results in this study indicate that the metabolic disturbance in PK deficiency alters not only the survival of red cells but also the maturation of erythroid progenitors, resulting in ineffective erythropoiesis. PMID- 16263414 TI - Erythropoietin-independent erythroid colony formation by bone marrow progenitors exposed to interleukin-11 and interleukin-8. AB - OBJECTIVE: Endogenous erythroid colonies (EECs), formed in vitro without erythropoietin (EPo) or other exogenous cytokines, are characteristic of Polycythemia vera (PV). Our aim was to identify specific conditions of culture of bone marrow (BM) progenitors allowing formation of erythroid colonies without EPo. METHODS: BM mononuclear cells (BMMCs), purified CD34+ cells, and purified CD36+ erythroid progenitors were cultured in serum-free media without and with cytokines: EPo, stem cell factor (SCF), and interleukin (IL)-11 and IL-8, produced by BM stromal cells and found elevated in PV. RESULTS: EECs were formed in PV cultures of either BMMCs or CD34+ cells, which include cytokine-secreting cells, but not in cultures of purified CD36+ erythroid progenitors (EP). Despite expression of V617F JAK-2, no constitutive activation of JAK-2, Stat-5, or Erk 1/2 was detected in erythroblasts issued from PV CD36+ progenitors. However, when SCF was provided, PV CD36+ progenitors formed erythroid colonies without EPo. The ability to form erythroid colonies with SCF alone was conferred to BM progenitors of healthy donors and secondary erythrocytosis by exposure to IL-11 and IL-8. Both IL-11 and IL-8 enhanced formation of erythroid colonies in response to EPo and interfered with the activation of Erk-1/2 and Stat-5 induced, respectively, by SCF and EPo in erythroblasts. Anti-IL-11 antibody inhibited formation of erythroid colonies by PV BMMCs and CD34+ cells. CONCLUSION: The data indicate that PV erythroid progenitors remain cytokine-dependent and that normal BM progenitors exposed to IL-11 and IL-8 can acquire the ability to form erythroid colonies without EPo. PMID- 16263415 TI - HDL cholesterol levels are an important factor for determining the lifespan of erythrocytes. AB - OBJECTIVE: Scavenger receptor class B, type I (SR-BI) is a multifunctional receptor that promotes the selective uptake of cholesteryl esters from high density lipoprotein (HDL). Disruption of SR-BI in mice results in a dramatic increase in HDL cholesterol. Interestingly, mice lacking SR-BI also develop anemia, as evidenced by accumulation of reticulocytes in the circulation. The objective of the current study was to delineate the mechanism underlying development of anemia in the absence of SR-BI. METHODS: Expression of important mediators of erythropoiesis, as well as key enzymes in the degradation of erythrocytes, were analyzed using real-time polymerase chain reaction in SR-BI wild-type and SR-BI knockout mice. In addition, in vivo studies were performed using biotinylated erythrocytes to determine erythrocyte survival. RESULTS: mRNA expression of TAL-1, GATA-1, FOG-1, erythropoietin receptor, and ferrochelatase, important mediators of erythropoiesis, was increased in spleens of SR-BI deficient mice. In addition, the relative amount of early Ter119(high)CD71(high) expressing erythroblasts was increased in SR-BI-deficient spleens. Interestingly, also expression of hemeoxygenase 1 and biliverdin reductase, enzymes involved in the degradation of erythrocytes, was increased. Furthermore, an elevated amount of conjugated bilirubin, the breakdown product of hemoglobin, was found in bile. Using biotinylated erythrocytes, we show that survival of erythrocytes was decreased in SR-BI-deficient mice. Thus, the observed increased erythropoiesis in the SR-BI-deficient mice is most likely a direct response to the reduced erythrocyte lifespan. Finally, we show that increased HDL cholesterol levels due to SR-BI deficiency induce erythrocyte cholesterol:phospholipid ratios, resulting in decreased deformability and increased osmotic fragility, thereby providing an explanation for the observed reduced lifespan. CONCLUSIONS: SR-BI is not only essential for HDL cholesterol homeostasis and atherosclerosis susceptibility, but also for maintaining normal erythrocyte lifespan. PMID- 16263416 TI - Efficient gene transfer into primary human natural killer cells by retroviral transduction. AB - OBJECTIVE: To optimize retroviral gene transfer into primary human natural killer (NK) cells. MATERIALS AND METHODS: NK cells from healthy donors were expanded ex vivo for a period of 21 days. Retroviral transductions were carried out by replacing culture media with retrovirus-containing supernatant during 2-hour incubations on days 3, 4, 5, 6, 10, 15, or 20. In some experiments, NK cells were transduced on 2 consecutive days (days 5 and 6). Green fluorescent protein served as a marker for detection of transduced cells. RESULTS: NK cells showed a median of 27.2% transduction efficiency after a single transduction round (transduction on day 5) and a median of 47.1% transduction efficiency after two rounds of transduction (transduction on days 5 and 6), 24 hours after exposure to retrovirus-containing supernatants. On day 21 after initial culture, 51.9% of NK cells were transduced after a single transduction round (transduction on day 5) and 75.4% after two rounds of transduction (transduction on days 5 and 6). Gene transfer did not change the function or phenotype of NK cells as determined by phenotypical analysis, nor did the proliferative ability or cytotoxic function change. CONCLUSION: The results show that NK cells can successfully be transduced with retroviral vectors, without any detectable changes in phenotype or function. This may open up new possibilities in the studies of NK cell biology and the development of NK cells for immunotherapy regimens. PMID- 16263417 TI - The N-terminal tetrapeptide of neutrophil proteinase 3 causes S-phase arrest in granulopoietic progenitors. AB - OBJECTIVE: Secreted enzymatically inactive proforms of hematopoietic serine proteases proteinase 3 (PR3), azurocidin, and granzymes A, B, H, K, and M are able to reduce the fraction of granulopoietic progenitors (CFU-GM) in S-phase, whereas human leukocyte elastase (HLE) and cathepsin G lack this ability. The objective of the present study was to map the specific sequence(s) of PR3 and other hematopoietic serine proteases responsible for the downmodulation of S phase. METHODS: Synthetic peptides corresponding to N-terminal sequences of PR3, purified recombinant PR3, and HLE, as well as hybrid proteins constructed by interchanging the N-terminal regions of PR3 and HLE, thus creating PR3/HLE and HLE/PR3, respectively, were tested for their ability to reduce the fraction of human marrow CFU-GM killed by cytosine arabinoside. In addition, we measured the effect of synthetic peptides on bromodeoxyuridine (BrdU) incorporation in common myeloid progenitors (CMP) and granulocyte/macrophage progenitors (GMP) isolated by cell sorting. RESULTS: The common N-terminal motif of PR3 and other serine proteases (i.e., IVGG or IIGG) downmodulate the S-phase of CFU-GM at 40 to 80 nM concentration. Tetrapeptide IVGG, but not IVGR, significantly reduces BrdU incorporation in GMP within the CD34+ population. When the N-terminal of HLE is presented by the HLE/PR3 hybrid protein it is fully active. CONCLUSION: These findings demonstrate that the downmodulatory effect on CFU-GM in S-phase is an S phase arrest mediated by the first four N-terminal amino acids of PR3, and also suggest that this activity is dependent on the configuration of the proform providing the correct presentation of this N-terminal motif. PMID- 16263418 TI - Adenovirus infection and cytotoxicity of primary mantle cell lymphoma cells. AB - Mantle cell lymphoma (MCL) is a distinct form of non-Hodgkin's lymphoma (NHL) derived from CD5+ B cells. MCL cells overexpress cyclin D1 as a consequence of translocation of the gene into the immunoglobulin heavy-chain gene locus. MCL is an aggressive form of NHL with frequent relapses after standard-dose chemotherapy. In this context, a variety of novel therapies for patients with MCL have been investigated. In this study, we use an expanded panel of attenuated adenoviruses to study adenovirus-mediated cytotoxicity of MCL cells. Our results demonstrate: 1) adenovirus infection of MCL cells despite the absence of receptor/coreceptor molecules known to be important for adenovirus infection of other cells types; 2) cytotoxicity of MCL cells after infection with specific adenovirus mutants; 3) a high degree of cytotoxicity after infection of some patient samples with viruses lacking the E1B 19k "antiapoptotic" gene; and 4) cytotoxicity after infection with viruses containing mutations in E1A pRb or p300 binding. The extent of cytotoxicity with the panel of viruses demonstrated interpatient variability, but 100% cytotoxicity, as determined by molecular analysis, was detected in some samples. These studies provide the foundation for: 1) the development of adenoviruses as cytotoxic agents for MCL and 2) analyses of key regulatory pathways operative in MCL cells. PMID- 16263419 TI - Inhibitory effects of tumor necrosis factor on hematopoiesis seen in vitro are translated to increased numbers of both committed and multipotent progenitors in TNF-deficient mice. AB - OBJECTIVES: The effects of TNF deficiency on myelopoiesis were evaluated in long term (LTBMC) and short-term bone marrow cultures (STBMC) and compared to hematopoietic activity in vivo in TNF-deficient mice. METHODS: LTBMC and STBMC were established from bone marrow of TNF-deficient mice in the presence or absence of soluble TNF. Total cell production was measured over time, as well as the number of colony-forming units in culture (CFU-C). Morphology of nonadherent (NA) cells in LTBMC was assessed after 10 weeks. Bone marrow cells (BMC) and peripheral blood (PB) cells were used to determine lineage distribution within the hematopoietic system. BMC were sorted to obtain Lin(-)c-kit(+)Sca-1- and Lin( )c-kit(+)Sca-1+ cells, which were plated in semisolid media to determine CFU-C numbers or injected into irradiated recipients to determine colony formation in the spleen (CFU-S). RESULTS: TNF-deficient LTBMC and STBMC show increased proliferative capacity, which can be inhibited by exogenous TNF to wild-type levels. Morphological analysis of NA cells from TNF-deficient LTBMC revealed increased numbers of cells at early stages of granulocytic differentiation (myeloblasts/promyelocytes) paralleled by a sharp decrease in the number of terminally differentiated polymorphonuclear neutrophils. Slightly elevated numbers of leukocytes, mainly neutrophils, were detected in PB of TNF-deficient mice. In bone marrow of TNF-deficient mice a significant increase in the number of both CFU-GM within Lin(-)c-kit(+)Sca-1- population and CFU-S within Lin(-)c kit(+)Sca-1+ population was observed. CONCLUSIONS: TNF has inhibitory effects on granulocyte-macrophage progenitors in vitro and on committed and primitive hematopoietic progenitors in vivo. However, in adult organism TNF deficiency is mostly compensated and controlled. PMID- 16263420 TI - Killer immunoglobulin-like receptor genotype in immune-mediated bone marrow failure syndromes. AB - OBJECTIVE: Recent reports have shown that killer immunoglobulin-like receptors (KIR) and KIR ligand (KIR-L) genotype play a role in the pathophysiology of autoimmune disorders. Certain KIR/KIR-L combinations might convey a predisposition to immune-mediated bone marrow failure. Examining the genotypic makeup of human leukocyte antigen (HLA) class I (KIR-L) and KIR in patients with hematologic disorders could offer clues to immunogenetic risk factors for these diseases. The objective of this study is to establish the frequency of specific KIR genes and KIR-L alleles in aplastic anemia (AA), paroxysmal nocturnal hemoglobinuria (PNH), myelodysplasia (MDS), and large granular lymphocyte leukemia (LGL), as compared with healthy control patients. METHODS: KIR genotyping was performed on DNA from 113 patients with AA, PNH, MDS, and LGL using sequence specific primer amplification. Results of genotypic KIR analysis were examined with HLA typing. Comparisons in frequency of KIR-L groups, KIR genotype profiles, and KIR/KIR-L mismatch were performed to determine whether specific KIR genes and KIR-L are overrepresented in bone marrow failure states. RESULTS: No difference in frequency of KIR-L groups was found relative to the healthy controls. AA and PNH showed decreased frequency of KIR-2DS1 and KIR-2DS5 genes: KIR-2DS1 35% vs 21% (p = 0.15) for AA and 19% for PNH (p = 0.13); KIR-2DS5 31% vs 14% (p = 0.08) for AA and 12% (p = 0.06) for PNH. These differences were even greater when compared to a larger group of control individuals from a study with similar genetic background. CONCLUSIONS: The reduced frequency of these KIR in AA and PNH may indicate an immunogenetic relationship between these diseases. PMID- 16263421 TI - Allele-specific transcription of fetal genes in primary erythroid cell cultures from Lepore and deltabeta degrees thalassemia patients. AB - OBJECTIVE: Autonomous gene silencing and gene competition by globin promoters for locus control region (LCR) function have been proposed as mechanisms in developmental regulation of beta-like genes. deltabeta degrees thalassemias are syndromes presenting an increased production of fetal hemoglobin in adult life; the majority of them are due to various deletions in beta-globin gene cluster. We studied samples from double heterozygotes for beta-thalassemia and for Lepore or Sicilian deltabeta degrees deletions, both lacking beta-promoter sequence. Our goal was to address the question of whether the allele carrying the deltabeta degrees deletion is responsible for high level of fetal hemoglobin (HbF) production. PATIENTS AND METHODS: We analyzed the globin gene transcription in human erythroid cell cultures from peripheral blood stem cells, using primary transcript in situ hybridization. We performed primary erythroid cultures from patients with the following genotypes: Lepore/beta degrees 39, Sicilian deltabeta degrees /beta degrees 39, and, as controls, two thalassemia patients with nondeletional mutations (IVS1,6/IVS1,6; IVS1,6/beta degrees 39), and one normal individual. RESULTS: The cells where it is possible to unambiguously assign gamma genes transcription in cis with the deletion (gamma:beta) are strongly represented with respect to the nine other combinations of gamma and beta hybridization signals. These cells are at least nine times more represented than those expressing the gamma allele in trans to the deletion. CONCLUSION: The allele-specific transcription of fetal genes in cis with the deletion is favored in both deletional genotypes. The absence of the adult promoter may influence LCR recruitment by fetal promoter, supporting the hypothesis that competition mechanism and gene silencing can coexist in regulating human globin gene transcription. PMID- 16263422 TI - Feasibility of cord blood stem cell manipulation with high-energy shock waves: an in vitro and in vivo study. AB - OBJECTIVE: Cord blood CD34+ cells are more uncommitted than their adult counterparts as they can be more easily maintained and expanded in vitro and transduced with lentiviral vectors. The aim of this study was to evaluate whether pretreatment with high-energy shock waves (HESW) could further enhance the expansion of cord blood progenitors and the transduction efficiency with lentiviral vectors. METHODS: Human cord blood CD34+ cells underwent HESW treatment with a wide range of energy and number of shots (from 0.22 mJ/mm2 to 0.43 mJ/mm2 and from 200 to 1500 shots). Cells were then evaluated both for their in vitro expansion ability and in vivo engraftment in primary, secondary, and tertiary NOD/SCID mice. The transduction efficiency with a lentiviral vector (LV) was also evaluated in vitro and in vivo. RESULTS: Cell viability following HESW ranged from 75 to 92%. Pretreatment with HESW significantly improved early progenitor cell expansion after short-term suspension culture. Upon transplantation in primary NOD/SCID mice, the HESW treatment enhanced progenitor cell engraftment (total human CD45(+)CD34+ cells were 10% in controls and 14.5% following HESW, human CD45(+)CD34(+)CD38(-) cells were 0.87% in controls and 1.8% following HESW). HESW treatment enhanced the transduction of a GFP+ lentiviral vector (e.g., at day 42 of culture 6.5% GFP+ cells in LV-treated cell cultures compared to 11.4% of GFP+ cells in HESW-treated cell cultures). The percentage of human GFP+ cell engrafting NOD/SCID mice was similar (34% vs 26.4% in controls); however, the total number of human cells engrafted after HESW was higher (39.6% vs 15%). CONCLUSION: The pretreatment of CD34+ cells with HESW represents a new method to manipulate the CD34+ population without interfering with their ability to both expand and engraft and it might be considered as a tool for genetic approaches. PMID- 16263424 TI - Comparative characteristics of mesenchymal stem cells from human bone marrow, adipose tissue, and umbilical cord blood. AB - OBJECTIVE: Various preparative protocols have been proposed for the acquisition and cultivation of mesenchymal stem cells (MSC). Whereas surface antigen markers have failed to precisely define this population, microarray analysis might provide a better tool for characterization of MSC. METHODS: In this study, we have analyzed global gene expression profiles of human MSC isolated from adipose tissue (AT), from umbilical cord blood (CB), and from bone marrow (BM) under two growth conditions and have compared them to terminally differentiated human fibroblasts (HS68). Profiles were compared using our Human Genome Microarray representing 51.144 different cDNA clones. RESULTS: Cultured with the appropriate conditions, osteogenic and adipogenic differentiation could be confirmed in all MSC preparations but not in fibroblasts. No phenotypic differences were observed by flow cytometry using a panel of 22 surface antigen markers. Whereas MSC derived from different donors using the same culture procedure yielded a consistent and reproducible gene expression profile, many genes were differentially expressed in MSC from different ontogenetic sources or from different culture conditions. Twenty-five genes were overlapping and upregulated in all MSC preparations from AT, CB, and BM as compared to HS68 fibroblasts. These genes included fibronectin, ECM2, glypican-4, ID1, NF1B, HOXA5, and HOXB6. Many genes upregulated in MSC are involved in extracellular matrix, morphogenesis, and development, whereas several inhibitors of the Wnt pathway (DKK1, DKK3, SFRP1) were highly expressed in fibroblasts. CONCLUSION: Our results have provided a foundation for a more reproducible and reliable quality control using genotypic analysis for defining MSC. PMID- 16263423 TI - Characterization of primitive marrow CD34+ cells that persist after a sublethal dose of total body irradiation. AB - Knowledge of the molecular events that occur during hematopoietic stem/progenitor cell (HSPC) development is vital to our understanding of blood cell production. To study the functional groups of genes characteristic of HSPCs we isolated a subpopulation of CD34+ bone marrow (BM) cells from nonhuman primates that persisted in vivo after a sublethal dose of total body irradiation (TBI). CD34+ cells isolated during the phase of maximal hematopoietic suppression show a transcriptional profile characteristic of metabolically inactive cells, with strong coordinate downregulation of a large number of genes required for protein production and processing. Consistent with this profile, these CD34+ cells were not able to generate hematopoietic colonies. Transcriptional profiling of these CD34+ cells in conjunction with a pathway analysis method reveals several classes of functionally related genes that are upregulated in comparison to the CD34+ cells obtained prior to TBI. These families included genes known to be associated with self-renewal and maintenance of HSPCs (including bone morphogenetic proteins), resistance to apoptosis (Bcl-2) as well as genes characteristic of a variety of nonhematopoietic tissues (gamma-aminobutyric acid/glycine receptor, complement receptor C1qRp). In contrast, during the period of hematopoietic recovery, the CD34+ cells expressed higher level of genes encoding factors regulating maturation and differentiation of HSPCs. Our data indicate that the primitive BM CD34+ cell population that persists after radiation possesses a transcriptional profile suggestive of pluripotency. PMID- 16263425 TI - Intra-bone marrow transplantation facilitates pauci-clonal human hematopoietic repopulation of NOD/SCID/beta2m(-/-) mice. AB - OBJECTIVE: Intra-bone marrow transplantation (IBMT) has been shown to improve the limit of detection of primitive human SCID-repopulating cells (SRC) in NOD/SCID mice when compared to intravenous transplantation. We sought to further refine detection of SRC by comparing NOD/SCID mice to the more sensitive NOD/SCID/beta2m(-/-)strain as IBMT recipients of limiting numbers of purified primitive human hematopoietic cells. MATERIALS AND METHODS: Purified human Lin( )CD34(+)CD38- cells at limiting doses were delivered by IBMT into NOD/SCID and NOD/SCID/beta2m(-/-) strains of recipient mice. Six weeks posttransplantation, injected and noninjected bones were analyzed separately for multilineage human hematopoietic chimerism. RESULTS: NOD/SCID/beta2m(-/-) mice are superior recipients for IBMT and show a trend toward increased levels of human hematopoietic engraftment. In addition, in contrast to NOD/SCID recipients, NOD/SCID/beta2m(-/-) mice were reconstituted with as few as five highly purified cells, indicative of pauci-clonal repopulation. Analysis of injected and noninjected bones demonstrated that engrafting cells were capable of in vivo migration and expansion. Although SRC hematopoietic reconstitution of NOD/SCID mice is commonly lymphoid-dominant, multilineage analysis of separate bone sites following IBMT of purified cells revealed that a subset of mice was repopulated with a myeloid-dominant graft in at least one bone site, revealing that SRC are developmentally heterogeneous among Lin(-)CD34(+)CD38- cells and capable of distinct differentiation potential. CONCLUSION: IBMT into NOD/SCID/beta2m(-/-) mice provides a highly sensitive experimental transplantation assay for the detection of human hematopoietic repopulating cells and demonstrates that Lin( )CD34(+)CD38- cells are more highly enriched for human SRC than originally predicted. PMID- 16263426 TI - Lactic acid bacteria and their uses in animal feeding to improve food safety. PMID- 16263427 TI - Mycotoxins in fruits: microbiology, occurrence, and changes during fruit processing. PMID- 16263428 TI - Human protozoan parasites in molluscan shellfish. PMID- 16263429 TI - Regulation of human immune and inflammatory responses by dietary fatty acids. PMID- 16263430 TI - Bacteria important during winemaking. PMID- 16263431 TI - Biosynthesis of conjugated linoleic acid in ruminants and humans. PMID- 16263432 TI - Safety issues associated with herbal ingredients. PMID- 16263435 TI - Epidemiology and bacteriology of bacterial pneumonias. AB - An understanding of the epidemiology and bacteriology of pneumonia in childhood is frustrated by the difficulties in making an aetiological diagnosis in young children. Incidence figures for community-acquired pneumonia are derived from the 1970s and 1980s and suggest that the incidence is higher in young children than in older children: approximately 40/1000/year <5 years of age and 15/1000/year aged 5-14 years. Certain pathogens are associated with pneumonia in different age groups, Mycoplasma pneumoniae and Chlamydia pneumoniae are more commonly implicated in older age groups and viruses in younger age groups. The most common bacterium, Streptococcus pneumoniae is less influenced by age. The mortality from community-acquired pneumonia in developed countries appears to be very low. The range of potential bacterial pathogens associated with nosocomial pneumonia and with pneumonia in the immunosuppressed child is wider with Gram-negative aerobic bacilli the most frequent isolate. PMID- 16263436 TI - Diagnosis and investigation of bacterial pneumonias. AB - This overview has been supported by a review of the literature referring to the management of community-acquired pneumonia (in preparation). Difficulties diagnosing bacterial pneumonia include identifying the pathogens and the validation of radiographic signs suggesting bacterial or mycoplasmal infection. The World Health Organisation (WHO) has published guidelines for diagnosis which seem to be as relevant for developed as developing countries. The main diagnostic features are tachypnoea, fever greater than 38.5 degrees C and chest recession without wheeze. Radiographic features lag behind clinical findings and provide little additional help when antibiotic treatment is considered but are crucial in the proper management of complications. PMID- 16263437 TI - Bacterial pneumonias: management and complication. AB - The treatment of bacterial pneumonia can occur either in the community or hospital setting depending on the clinical status of the child. In the community, supportive management includes anti-pyretics, analgesia and early review by a medical practitioner if there is deterioration or no response to oral antibiotics within 48 hours. Suitable first-line antibiotics in the under fives are penicillin- or cephalosporin-based and in the over fives macrolides may also be considered. In a severely ill child, good oxygenation is essential and oxygen may be delivered by mask, nasal prong or headbox. Persistent fever, despite appropriate treatment warrants further investigations. The most common complication is an empyema and treatment is effective drainage with continuing high-dose antibiotics. Other lung complications include abscess formation and pneumatoceles. The syndrome of inappropriate anti-diuretic hormone secretion occurs in approximately one third of hospitalized patients with pneumonia. A rare but important complication is pericarditis. PMID- 16263438 TI - Impact of bacterial pneumonias on world child health. AB - Lower respiratory infections (LRIs) are estimated to cause approximately 4 million deaths each year, the majority in previously healthy young children from low-income countries. Bacterial pneumonias are likely to account for the major proportion of deaths but because accurate diagnosis is difficult, their precise contribution is difficult to estimate. Effective protection against two of the most important pathogens (H. influenzae type B and S. pneumoniae) is now available even for young infants. The high cost of these interventions is however likely to prohibit their use in areas likely to benefit most. Case management which can be effective at reducing mortality and which is relatively cheap at present is likely therefore to remain the principal means of tackling bacterial pneumonias globally. Antibiotic resistance to cheap drugs and possibly HIV infection pose a potentially enormous threat to this strategy with the prospect of an increasing global death toll with an even more inequitable distribution of disease as we begin the new century. PMID- 16263440 TI - Local mucociliary defence mechanisms. AB - The lung is continually at risk of exposure to noxious environmental agents and respiratory pathogens. An elaborate series of defence mechanisms have been developed to protect the airways from these insults. The lower respiratory tract is protected by local mucociliary mechanisms that involve the integration of the ciliated epithelium, periciliary fluid and mucus. Mucus acts as a physical and chemical barrier onto which particles and organisms adhere. Cilia lining the respiratory tract propel the overlying mucus to the oropharynx where it is either swallowed or expectorated. Regulation of periciliary fluid is thought essential to maintenance of both mucociliary clearance and to produce an environment in which airway antimicrobial peptides and defensins are effective. Disruption of mucociliary clearance may be caused by diseases such as cystic fibrosis, primary ciliary dyskinesia and asthma or may be secondary to pollutant exposure and viral or bacterial infections. PMID- 16263442 TI - The physiology of forced expiration. AB - Insight into the physiology of a forced expiration is the most important prerequisite for understanding and correctly interpreting the most common pulmonary function tests like spirometry and recording of a maximum expiratory flow-volume curve. Along the airway, intraluminal pressure falls progressively from alveolar pressure in the periphery to atmospheric pressure at the airway opening. Downstream of the equal pressure point, where intraluminal equals transthoracic pressure, the airway is dynamically compressed. Once the forced expiration is flow limited, more transthoracic pressure will rather effect more compression than a further increase in flow. It follows that the achieved maximum expiratory flow rates are then exclusively defined by the resistance of the intrathoracic airways and no longer by muscular effort. The end of a forced expiration is determined by the elastic resistance of the thoracic cage and by airway closure. PMID- 16263445 TI - Bronchoalveolar lavage. AB - Bronchoalveolar lavage is a technique for sampling the epithelial lining fluid of the respiratory tract. Analysis of cellular and non-cellular components of returned fluid has the potential to provide valuable information about airways inflammation. Because of the invasive nature of the investigation, there are few conditions for which repeat sampling can be justified. Bronchoalveolar lavage has been used to study immune mechanisms in cystic fibrosis, interstitial lung diseases and asthma. This article reviews the usefulness of BAL assessments for lung inflammation in paediatric practice. PMID- 16263447 TI - The lung in multi-system disease. AB - Lung involvement in multi-system disease is rare in children. When it does occur it is associated with significant morbidity and mortality. Auto-immune diseases such as systemic lupus erythematosis, scleroderma and acute rheumatic disease can cause major lung pathology. Chronic illness such as sickle cell disease, mucopolysaccharidosis or familial dysautonomia can result in long-term recurrent respiratory problems. Rare conditions such as sarcoidosis, pulmonary lymphangiomatosis or Langerhan's histiocytosis occasionally result in major lung disease in this age group. The prognosis for all of these conditions is very variable depending on the severity of the disease itself and its response to treatment which often involves the use of oral corticosteroids or other powerful immuno-suppressants. PMID- 16263448 TI - Non cystic fibrosis bronchiectasis. AB - Bronchiectasis is becoming less common as the treatment for acute lower respiratory tract infections improves and immunization programmes decrease the frequency of pertussis and measles. However bronchiectasis is still a challenge to the paediatric chest physicians in many developing parts of the world and it remains a frequent problem being the final common pathway of several different lower respiratory tract insults such as cystic fibrosis, immunodeficiency, ciliary dyskinesia. Although the treatment of patients with bronchiectasis is primarily medical, surgical treatment is required in a small group of patients with recurrent episodes of pneumonia and atelectasis localized to one area, severe or recurrent hemoptysis and in those unresponsive to aggressive medical treatment with abnormal growth and development. There are unanswered questions about childhood bronchiectasis, mainly on aetiology and treatment which require more research. PMID- 16263449 TI - The changing picture of childhood asthma. AB - Although the last four decades have seen substantial progress in the understanding of the pathophysiology of asthma, the working clinician's main tool in establishing the diagnosis of asthma is a good and accurate clinical history. In the older child, the history is complemented by pulmonary function tests, including measures of bronchial lability. After nearly three decades of increasing hospital admissions for childhood asthma, the 1990s have seen a small reduction in overall admissions. The increase in hospital admissions is explained only partly by the rising prevalence of asthma. The associated morbidity due to the disease has increased quite considerably over the last three decades. International studies assessing asthma prevalence across different parts of the globe using identical methodologies has shown considerable differences in asthma prevalence between the East and the West as well as within countries. The burden of paediatric asthma on the child, the family and society is increasing as we move into the new millennium. PMID- 16263450 TI - Case 1: Assessment: chronic wet cough. PMID- 16263451 TI - Case 2: Assessment: chest X-ray. PMID- 16263457 TI - Lung growth for beginners. AB - Lung growth occurs as a series of tightly regulated events commencing in the embryo and continuing post-natally. It depends on a number of factors, including developmental, genetic and environment ones. Abnormalities of any of these factors may have a profound influence on lung growth. The causes of developmental abnormalities of the lung such as lung cysts and congenital diaphragmatic hernia are poorly understood, but may result from a combination of genetic and environmental factors. Normal fetal breathing movements and an adequate balance between the production of fetal lung fluid and drainage of this fluid are both essential for normal fetal lung growth. It seems that fetal breathing movements are necessary to maintain sufficient pressure within the airways and perhaps to directly stimulate lung growth via induction of mitogenic activity. The volume of intra-pulmonary fluid is regulated by the resistance of the upper airway and by contractions of the diaphragm. Increased drainage of the amniotic fluid, another essential factor for normal lung growth and development, will result in marked pulmonary hypoplasia as may occur with pre-term rupture of the membranes and with fetal renal disease. Perhaps the most important factor for adverse lung growth is pre-term delivery of the infant from any cause including intra-uterine infection. Both ante- and post-natal factors, including mechanical ventilation and oxygen therapy, will affect normal alveolization. In this review, particular attention is paid to breathing movements and the balance between fluid production and drainage. PMID- 16263458 TI - Factors adversely affecting lung growth. AB - Antenatal lung growth is adversely affected in conditions where there is intra-or extrathoracic compression, impaired fetal breathing movements and/or reduction in amniotic fluid volume. Postnatally, factors in infancy most commonly associated with abnormal lung function are passive smoking and premature delivery, its associated complications and management. Many conditions affecting lung growth are readily diagnosed by antenatal ultrasonography. Postnatally, pulmonary hypoplasia should be suspected if an infant requires high airway pressures during resuscitation and subsequent ventilatory support, in the absence of severe parenchymal disease on the chest radiograph. Antenatal therapy has been directed at relieving lung compression, but the efficacy of such interventions has not been proven in randomized trials. Certain disorders associated with impaired lung growth have a high perinatal mortality. Infants who are less severely affected suffer respiratory symptoms with lung function abnormalities or may only be distinguished by chronic tachypnoea during infancy. PMID- 16263459 TI - Growth of the vascular tree. AB - Major changes occur in the pulmonary circulation at birth. Failure of normal adaptation leads to pulmonary hypertension of the newborn. The basis for this may be abnormal growth of the lung in utero or a failure of the mechanisms which lead to vasodilatation in the newborn period. This article describes normal development of the pulmonary arteries and veins, their branching pattern and structure and discusses the factors which may control this growth. Endothelial function and dysfunction are crucial in adaptation, and new experimental studies are aiding our understanding of the control and action of factors produced by the endothelium, e.g. nitric oxide, prostacyclin and endothelin. The study of infants with pulmonary hypertension, together with the experimental studies, will aid in producing effective methods of controlling increased pulmonary artery pressure in the newborn period. PMID- 16263460 TI - Clinical outcomes of congenital lung abnormalities. AB - Individually, congenital abnormalities of the lung are rare but collectively they form an important group of conditions. The range of malformations is broad and the clinical impact is very variable, depending on the degree of malformation. This article groups these conditions under their traditional headings and considers their management. However, in conditions that are atypical, it is often useful to adopt a clear anatomical approach. Consider investigations that will clearly identify the structure of the bronchial tree, lung parenchyma and the vascular supply and drainage. Describe each in logical and simple terms. Similarly, an individual assessment of function will assist in planning appropriate management. The natural history of some of the congenital lung lesions is often unknown. This gap in our knowledge has widened with the advent of detailed antenatal ultrasound scanning. Paediatricians are now faced with an increasing cohort of completely asymptomatic antenatally diagnosed congenital lung lesions and the best post-natal management is unclear. Long-term follow-up studies are required to document their natural history. PMID- 16263461 TI - Upper airway defence mechanisms. AB - The nose is the airconditioner of the airways. Because normal breathing is through the nose, most airborne particles are filtered there; hence the nasal mucosa is the first line of defence against particles in the air. Pathogenic and non-pathogenic antigens continuously bombard the epithelium of the nasal airway. These antigens are mainly removed non-immunologically by the first defence layer of the mucosa, consisting of mucus, ciliated epithelial cells, glycoproteins/lysosymes. If the antigen passes this defence layer, specific and non-specific immunological defence mechanisms exist. The non-specific defence consists of phagocyting cells like neutrophils and macrophages and the complement activation. The specific defence mechanism (resulting in a specific immunological reaction in relation to a certain antigen) is formed by the antibodies, mainly secretory IgA and to a lesser extent IgG and immunocompetent cells in the nasal mucosa. Activation of the specific defence mechanisms may lead to inflammation which can be allergic. The intense co-operation of mechanical, aspecific and specific immunological defence results in a tightly controlled balance between a proper defence against pathogens and hypersensitivity. Failures in these defence mechanisms, or their co-operation, results in upper respiratory infection and/or allergy. PMID- 16263462 TI - Ethical issues in lung function testing in children. AB - Most lung function tests are non-invasive and they are widely performed in children and adults for clinical reasons. The prevalence of respiratory disorders is such that there is a considerable amount of research in the area. For both clinical and research applications, professionals involved in lung function testing in children need to be aware of ethical issues pertaining to such testing. In general, these are not specific to lung function testing but are common to other non-invasive procedures. These include the need to recognize the autonomy of the child (appropriate for age and maturity) and the importance of providing information in ways that a child can understand. The issues surrounding consent, confidentiality, and risk-benefit assessment are particularly important in vulnerable groups such as children. Parents, too, have a clear influence on healthcare issues surrounding children and hitherto have not always been appropriately involved. Healthcare professionals have a duty to children and their families to safeguard their psychological, social and emotional well-being, as well as their physical health. PMID- 16263463 TI - Ventilation and perfusion scanning in children. AB - Ventilation and perfusion scintigraphy (VQ scans) provide a relatively non invasive evaluation of lung function. They indicate the relative blood flow to each lung and allow a quantitative assessment of the perfusion of each lung segment. They can detect areas of abnormal aeration and demonstrate air trapping. However, VQ scanning gives relatively poor anatomical detail of the lungs and so is commonly used in conjunction with other radiological modalities, most notably the chest X-ray. VQ scans have been utilized for a large range of pathological conditions including congenital cardiac and lung abnormalities, suspected pulmonary emboli, the assessment of childhood diseases, including cystic fibrosis, bronchopulmonary dysplasia and asthma. PMID- 16263464 TI - Surrogate markers of airway inflammation: inflammometry in paediatric respiratory medicine. AB - Until recently, there has not been any practical way to assess airway inflammation non-invasively in paediatrics. Surrogate markers of airway inflammation are potentially of great importance in the diagnosis and monitoring of inflammatory airways disease in children. A large number of substances in blood, urine and exhaled air or induced sputum are currently under study to evaluate their possible usefulness as markers of airway inflammation. To be useful, a marker should be valid, preferably non-invasive, quick, reproducible, repeatable and cheap. In addition, markers should be studied in relation to their specific purpose because different markers may be useful for different types of airway inflammation. Few, if any, markers will fulfill all these requirements. Most research has focused on applications of markers in asthma, some data refer to cystic fibrosis, infections and ciliary dyskinesia. Of all surrogate markers, exhaled nitric oxide has been studied the most and seems to offer information that should be evaluated for its relevance to clinical practice. Before introducing markers of inflammation into daily practice, analysis of benefits and costs are needed. There is little doubt that 'inflammometry' will be a major step forward and will be useful in differentiating airways diseases and improving treatment. PMID- 16263465 TI - Pulmonary hypertensive diseases. AB - The commonest causes of pulmonary hypertension are secondary to endstage pulmonary disease or congenital heart disease (including structural abnormalities of the pulmonary veins). Less obvious causes include sleep disordered breathing due to obstructive sleep apnoea or neuromuscular disease, and occult interstitial lung disease. When these have been excluded, the primary pulmonary vascular diseases should be considered. These are primary pulmonary hypertension; pulmonary veno-occlusive disease; pulmonary embolic disease (thromboembolism, and non-thrombotic embolism) and invasive pulmonary capillary haemangiomatosis. The clinical signs and chest X-ray appearances are often non-specific. Echocardiography can often estimate pulmonary artery pressure and exclude congenital heart disease. Right heart catheterization is usually needed to confirm the diagnosis, estimate any reversibility of elevated pulmonary vascular resistance and exclude other causes. Precise diagnosis may require an open lung biopsy. For many of these conditions, treatment is difficult and the prognosis poor unless the child has a lung transplant. PMID- 16263466 TI - Early use of inhaled corticosteroids in infancy. AB - The inability to accurately predict the outcome of infants with recurrent wheezy bronchitis makes the early use of inhaled corticosteroids (ICS) controversial. Data from bronchoalveolar lavages and epidemiological surveys suggest a persistent inflammation of the airways in the more severe cases. Prospective studies, mostly with nebulized corticosteroids, have demonstrated clinical efficacy on daytime and nightime symptoms, reduced requirements for rescue bronchodilators and a real steroid sparing effect. In infants with episodic viral associated wheeze with or without interval symptoms, ICS use carries the risk of overtreatment and of adverse effects. Long-term prospective studies are urgently required to assess the efficacy and safety of ICS and their possible effects on the natural history of infantile asthma. PMID- 16263475 TI - Acute lung injury: cellular mechanisms and derangements. AB - The clinical course of acute lung injury (ALI) is a complex and variable process accompanied by severe lung dysfunction, which persists for a long period of time with variable recovery of pulmonary function. The extent and severity of the lung disease associated with ALI varies with those patients having the most severe manifestations of lung disease being grouped as acute respiratory distress syndrome (ARDS). The pathological injury associated with this disease process, termed diffuse alveolar damage (DAD), has three overlapping phases (exudative, proliferative and fibrotic) which are the consequences of severe injury to the alveolar-capillary unit. There is no uniformity to the progression and length of each stage. This review explores those cellular mechanisms and derangements involved in the progression of ARDS. Those areas that demonstrate the major advances within the field are highlighted because of the diverse and vast nature of the cellular components involved in the process of ALI. We are beginning to identify those processes that contribute to the cellular derangements which are the hallmark of ALI. By expanding our understanding of those factors, we should in the future be able to construct therapeutic interventions that address the aetiology of ALI. PMID- 16263471 TI - Case 1: Assessment: symptomatic asthma. PMID- 16263477 TI - Acute lung injury: outcomes and new therapies. AB - Mortality rates in ARDS are improving, with several recent studies reporting mortality in the order of 20-40% rather than the early descriptions of this disease in which a mortality of 40-60% or higher was frequently cited. The ability to accurately predict outcomes plays an important role in the assessment of the impact of new therapies. Traditionally clinicians have relied on simple respiratory indices to assess mortality risk; however, the predictive ability of such indices, particularly early in the course of the disease, is somewhat limited. Adult data suggest that improved prediction not only of the outcome of established ARDS but also of the development of ARDS in at-risk patients may be obtained by measuring the concentrations of inflammatory mediators and/or surfactant-associated proteins in plasma or bronchoalveolar lavage samples. A bewildering array of therapies for ARDS is available; in many cases the benefits are uncertain. Treatments of proven value in adults include using PEEP beyond the lower inflection point of the pressure-volume curve and limiting tidal volumes to 6 ml/kg. Nitric oxide appears to offer no benefit to outcomes, although it does improve oxygenation in some patients. Surfactant is still undergoing assessment in randomised controlled trials; however, the use of aerosolised surfactant has been recently shown to be ineffective in adult patients with ARDS. Perfluorocarbon-assisted gas exchange (PAGE) or partial liquid ventilation is similarly still being assessed in randomised controlled trials in adults. PMID- 16263476 TI - Acute lung injury: pathophysiology, assessment and current therapy. AB - Acute respiratory distress syndrome (ARDS) is a clinically defined entity describing the severity of diffuse alveolar injury caused by direct or indirect injury to the lung. Pathophysiology, clinical course and outcome of ARDS depend on the underlying cause, the severity of the disease and co-morbidities. Pulmonary function tests show restrictive lung disease, which is characterised by a reduction in lung compliance and functional residual capacity, resulting in marked ventilation-perfusion inequality. Current ventilator strategies aim to minimise ventilator-induced lung injury by targeting mechanical ventilation between the lower and upper inflection point of the pressure volume curve. This includes recruitment manoeuvres and the use of high PEEP to open the atelectatic lung and the use of permissive hypercapnia and the limitation of peak inspiratory pressure below 35 cm H2O to avoid overinflation. The clinical benefit of newer modes of ventilatory support such as inverse ratio ventilation, high frequency oscillatory ventilation, surfactant replacement, prone positioning and inhaled nitric oxide has yet to be determined in children. PMID- 16263479 TI - Lung function tests for pre-school children. AB - There are a few devices for measuring lung function in pre-school children. Neither the interrupter technique nor the forced oscillation techniques have been standardised. We highlight some of the issues around the measurement of airway resistance using the interrupter technique and emphasise that, as with other lung function measurements, operators should have a proper understanding of the methods before they can be applied. Both methods for measuring airway resistance have potential for clinical and research application. PMID- 16263478 TI - Immunological investigations in children with recurrent respiratory infections. AB - Respiratory tract infections are common diseases in childhood. Most children with recurrent respiratory infections do not have an immunodeficiency. If they do, this is often due to an antibody deficiency. An important point in the investigation of a child with recurrent respiratory infections is to assess whether the child is thriving. If not, an underlying disease should be sought. Immunological investigations are useful if other, more frequent, underlying diseases have been ruled out. Early immunological screening is mandatory if there is a family history of immunodeficiency. In this review, a protocol is described which identifies children with severe antibody deficiency by simple screening tests before recurrent infections have caused irrepairable damage to the lungs. More elaborate tests are used to detect milder antibody deficiencies. These are reserved for those children in whom symptoms persist. PMID- 16263480 TI - Imaging the upper airways. AB - Radiological evaluation of the airway has been used as a screening tool and an adjunct to endoscopy for many years. It provides non-invasive data on the structure of the airway, often avoiding the risk of general anaesthesia. Standard radiographs provide some information on the intricate anatomy of the paediatric airway aided by fluoroscopy. More recently, CT and MRI are proving to have a valuable role and are approaching near endoscopic detail of airway anatomy. The purpose of this article is to highlight areas where radiology can aid in the evaluation of the airway of infants and children. PMID- 16263481 TI - Beta2-adrenoceptors: mechanisms of action of beta2-agonists. AB - The human beta2-adrenoceptor is a member of the 7 transmembrane family of receptors. It is encoded by a gene on chromosome 5 and is widely distributed in the respiratory tract. Following beta2-adrenoceptor activation, intracellular signalling is mainly produced by inducing cyclic AMP. This produces airway relaxation through phosphorylation of muscle regulatory proteins and modification of cellular Ca2+concentrations. Beta2-agonists have been characterised into those which directly activate the receptor (salbutamol/terbutaline), those which are taken up into a membrane depot (formoterol) and those which interact with a receptor-specific, auxiliary binding site (salmeterol). These differences in mechanism of action are reflected in the kinetics of airway smooth muscle relaxation and bronchodilation in asthmatic patients. Beta-adrenoceptor desensitisation is associated with beta2-agonist activation and differs depending on the cell type. It is reflected in the different profiles of clinical tolerance to chronic beta2-agonist therapy. A number of polymorphisms of the beta2-receptor have been described which appear to alter the behaviour of the receptor, including the degree of downregulation and response to beta2-agonists. PMID- 16263482 TI - Rare single system diseases. AB - There are many rare conditions to be found in textbooks of respiratory paediatrics which are often difficult to place in to any standard form of classification. The expertise of any single paediatrician or centre in the management of such problems is inevitably limited, and so treatment is never "evidence based" but depends on individual experience and information contained in case reports or small series in the literature. The investigation and management of such cases is often complex and should be performed in large regional centres. In this article we will discuss pulmonary alveolar microlithiasis, disorders of the lymphatic system, lymphoid bronchiolitis and pulmonary langerhans cell histiocytosis. PMID- 16263483 TI - The role of the collectin system in pulmonary defence. AB - The human collectin system comprises the serum protein, mannose- binding lectin and the hydrophilic surfactant proteins A and D. The three proteins possess structural and functional similarities and are important components of innate immunity. Through a variety of mechanisms, including direct opsonisation and complement activation, they assist in host defence against a wide array of micro organisms. Investigation of the roles of the surfactant proteins in pulmonary disease has been assisted recently by the development of transgenic knockout mice. Animals deficient in these proteins display susceptibility to certain bacterial and viral pathogens, stimulating research into the role of polymorphisms in these genes in human respiratory disease. The role of MBL in human pulmonary disease is less well established, although accumulating evidence suggests that it is a modifier for lung disease in tuberculosis and cystic fibrosis. PMID- 16263484 TI - The role of the nurse in paediatric asthma management. AB - In recent years the role that specialist nurses play in chronic disease management has broadened and they are now recognised as valuable members of the multi-disciplinary team, demonstrating that their skilled practice contributes to the quality of patient care. Nurses in primary and secondary care are working as autonomous practitioners managing their own caseload of asthmatic patients and taking a key role in facilitating ongoing treatment regimes. The development of advanced nursing roles has provided an opportunity to broaden many nursing skills producing highly competent paediatric respiratory nurses. The purpose of this article is to outline the service that nurses offer to children with asthma and their families by discussing the role they play in patient care and management. PMID- 16263487 TI - Case 1: assessment. Persistent granular lung fluid in a baby with TAPVD. PMID- 16263488 TI - Nuclear power and public health. PMID- 16263489 TI - The NIEHS responds to Hurricane Katrina. PMID- 16263490 TI - Occupational carcinogens: ELF MFs. PMID- 16263491 TI - The NAS perchlorate review: second-guessing the experts. PMID- 16263492 TI - In disaster's wake: tsunami lung. PMID- 16263493 TI - Global Earth observations for health. PMID- 16263494 TI - Power surge: renewed interest in nuclear energy. PMID- 16263495 TI - Harvesting the potential of biomass. PMID- 16263496 TI - Microbe power! PMID- 16263498 TI - Hypertension and Exposure to Noise near Airports (HYENA): study design and noise exposure assessment. AB - An increasing number of people live near airports with considerable noise and air pollution. The Hypertension and Exposure to Noise near Airports (HYENA) project aims to assess the impact of airport-related noise exposure on blood pressure (BP) and cardiovascular disease using a cross-sectional study design. We selected 6,000 persons (45-70 years of age) who had lived at least 5 years near one of six major European airports. We used modeled aircraft noise contours, aiming to maximize exposure contrast. Automated BP instruments are used to reduce observer error. We designed a standardized questionnaire to collect data on annoyance, noise disturbance, and major confounders. Cortisol in saliva was collected in a subsample of the study population (n = 500) stratified by noise exposure level. To investigate short-term noise effects on BP and possible effects on nighttime BP dipping, we measured 24-hr BP and assessed continuous night noise in another subsample (n = 200). To ensure comparability between countries, we used common noise models to assess individual noise exposure, with a resolution of 1 dB(A). Modifiers of individual exposure, such as the orientation of living and bedroom toward roads, window-opening habits, and sound insulation, were assessed by the questionnaire. For four airports, we estimated exposure to air pollution to explore modifying effects of air pollution on cardiovascular disease. The project assesses exposure to traffic-related air pollutants, primarily using data from another project funded by the European Union (APMoSPHERE, Air Pollution Modelling for Support to Policy on Health and Environmental Risks in Europe). PMID- 16263499 TI - Genetic factors that might lead to different responses in individuals exposed to perchlorate. AB - Perchlorate has been detected in groundwater in many parts of the United States, and recent detection in vegetable and dairy food products indicates that contamination by perchlorate is more widespread than previously thought. Perchlorate is a competitive inhibitor of the sodium iodide symporter, the thyroid cell-surface protein responsible for transporting iodide from the plasma into the thyroid. An estimated 4.3% of the U.S. population is subclinically hypothyroid, and 6.9% of pregnant women may have low iodine intake. Congenital hypothyroidism affects 1 in 3,000 to 1 in 4,000 infants, and 15% of these cases have been attributed to genetic defects. Our objective in this review is to identify genetic biomarkers that would help define subpopulations sensitive to environmental perchlorate exposure. We review the literature to identify genetic defects involved in the iodination process of the thyroid hormone synthesis, particularly defects in iodide transport from circulation into the thyroid cell, defects in iodide transport from the thyroid cell to the follicular lumen (Pendred syndrome), and defects of iodide organification. Furthermore, we summarize relevant studies of perchlorate in humans. Because of perchlorate inhibition of iodide uptake, it is biologically plausible that chronic ingestion of perchlorate through contaminated sources may cause some degree of iodine discharge in populations that are genetically susceptible to defects in the iodination process of the thyroid hormone synthesis, thus deteriorating their conditions. We conclude that future studies linking human disease and environmental perchlorate exposure should consider the genetic makeup of the participants, actual perchlorate exposure levels, and individual iodine intake/excretion levels. PMID- 16263500 TI - Personal exposure to ultrafine particles and oxidative DNA damage. AB - Exposure to ultrafine particles (UFPs) from vehicle exhaust has been related to risk of cardiovascular and pulmonary disease and cancer, even though exposure assessment is difficult. We studied personal exposure in terms of number concentrations of UFPs in the breathing zone, using portable instruments in six 18-hr periods in 15 healthy nonsmoking subjects. Exposure contrasts of outdoor pollution were achieved by bicycling in traffic for 5 days and in the laboratory for 1 day. Oxidative DNA damage was assessed as strand breaks and oxidized purines in mononuclear cells isolated from venous blood the morning after exposure measurement. Cumulated outdoor and cumulated indoor exposures to UFPs each were independent significant predictors of the level of purine oxidation in DNA but not of strand breaks. Ambient air concentrations of particulate matter with an aerodynamic diameter of < or = 10 microm (PM10), nitrous oxide, nitrogen dioxide, carbon monoxide, and/or number concentration of UFPs at urban background or busy street monitoring stations was not a significant predictor of DNA damage, although personal UFP exposure was correlated with urban background concentrations of CO and NO2, particularly during bicycling in traffic. The results indicate that biologic effects of UFPs occur at modest exposure, such as that occurring in traffic, which supports the relationship of UFPs and the adverse health effects of air pollution. PMID- 16263501 TI - Distribution of brevetoxin (PbTx-3) in mouse plasma: association with high density lipoproteins. AB - We investigated the brevetoxin congener PbTx-3 to determine its distribution among carrier proteins, including albumin and blood lipoproteins. Using a radiolabeled brevetoxin tracer (PbTx-3), we found that 39% of the radiolabel remained associated with components in mouse plasma after > 15 kDa cutoff dialysis. Of this portion, only 6.8% was bound to serum albumin. We also examined the binding of brevetoxin to various lipoprotein fractions. Plasma, either spiked with PbTx-3 or from mice treated for 30 min with PbTx-3, was fractionated into different-sized lipoproteins by iodixanol gradient ultracentrifugation. Each fraction was then characterized and quantified by agarose gel electrophoresis and brevetoxin radioimmunoassay, respectively. In both the in vitro and in vivo experiments, the majority of brevetoxin immunoreactivity was restricted to only those gradient fractions that contained high-density lipoproteins (HDLs). Independent confirmation of brevetoxin binding to HDLs was provided by high molecular weight (100 kDa cutoff) dialysis of [3H]PbTx-3 from lipoprotein fractions as well as a scintillation proximity assay using [3H]PbTx-3 and purified human HDLs. This information on the association of brevetoxins with HDLs provides a new foundation for understanding the process by which the toxin is delivered to and removed from tissues and may permit more effective therapeutic measures to treat intoxication from brevetoxins and the related ciguatoxins. PMID- 16263502 TI - Proximity to pollution sources and risk of amphibian limb malformation. AB - The cause of limb deformities in wild amphibian populations remains unclear, even though the apparent increase in prevalence of this condition may have implications for human health. Few studies have simultaneously assessed the effect of multiple exposures on the risk of limb deformities. In a cross sectional survey of 5,264 hylid and ranid metamorphs in 42 Vermont wetlands, we assessed independent risk factors for nontraumatic limb malformation. The rate of nontraumatic limb malformation varied by location from 0 to 10.2%. Analysis of a subsample did not demonstrate any evidence of infection with the parasite Ribeiroia. We used geographic information system (GIS) land-use/land-cover data to validate field observations of land use in the proximity of study wetlands. In a multiple logistic regression model that included land use as well as developmental stage, genus, and water-quality measures, proximity to agricultural land use was associated with an increased risk of limb malformation (odds ratio = 2.26; 95% confidence interval, 1.42-3.58; p < 0.001). The overall discriminant power of the statistical model was high (C = 0.79). These findings from one of the largest systematic surveys to date provide support for the role of chemical toxicants in the development of amphibian limb malformation and demonstrate the value of an epidemiologic approach to this problem. PMID- 16263503 TI - Framing scientific analyses for risk management of environmental hazards by communities: case studies with seafood safety issues. AB - Risk management provides a context for addressing environmental health hazards. Critical to this approach is the identification of key opportunities for participation. We applied a framework based on the National Research Council's (NRC) analytic-deliberative risk management dialogue model that illustrates two main iterative processes: informing and framing. The informing process involves conveying information from analyses of risk issues, often scientific, to all parties so they can participate in deliberation. In the framing process, ideas and concerns from stakeholder deliberations help determine what and how scientific analyses will be carried out. There are few activities through which affected parties can convey their ideas from deliberative processes for framing scientific analyses. The absence of participation results in one-way communication. The analytic-deliberative dialogue, as envisioned by the NRC and promoted by the National Institute of Environmental Health Sciences (NIEHS), underscores the importance of two-way communication. In this article we present case studies of three groups--an Asian and Pacific Islander community coalition and two Native American Tribes--active in framing scientific analyses of health risks related to contaminated seafood. Contacts with these organizations were established or enhanced through a regional NIEHS town meeting. The reasons for concern, participation, approaches, and funding sources were different for each group. Benefits from their activities include increased community involvement and ownership, better focusing of analytical processes, and improved accuracy and appropriateness of risk management. These examples present a spectrum of options for increasing community involvement in framing analyses and highlight the need for increased support of such activities. PMID- 16263504 TI - Associations of uric acid with polymorphisms in the delta-aminolevulinic acid dehydratase, vitamin D receptor, and nitric oxide synthase genes in Korean lead workers. AB - Recent research suggests that uric acid may be nephrotoxic at lower levels than previously recognized and that it may be one mechanism for lead-related nephrotoxicity. Therefore, in understanding mechanisms for lead-related nephrotoxicity, it would be of value to determine whether genetic polymorphisms that are associated with renal outcomes in lead workers and/or modify associations between lead dose and renal function are also associated with uric acid and/or modify associations between lead dose and uric acid. We analyzed data on three such genetic polymorphisms: delta-aminolevulinic acid dehydratase (ALAD), endothelial nitric oxide synthase (eNOS), and the vitamin D receptor (VDR). Mean (+/- SD) tibia, blood, and dimercaptosuccinic acid-chelatable lead levels were 37.2 +/- 40.4 microg/g bone mineral, 32.0+/- 15.0 g/dL, and 0.77+/- 0.86 microg/mg creatinine, respectively, in 798 current and former lead workers. Participants with the eNOSAsp allele had lower mean serum uric acid compared with those with the Glu/Glu genotype. Among older workers (age > or = median of 40.6 years), ALAD genotype modified associations between lead dose and uric acid levels. Higher lead dose was significantly associated with higher uric acid in workers with the ALAD1-1 genotype; associations were in the opposite direction in participants with the variant ALAD1-2 genotype. In contrast, higher tibia lead was associated with higher uric acid in those with the variant VDRB allele; however, modification was dependent on participants with the bb genotype and high tibia lead levels. We conclude that genetic polymorphisms may modify uric acid mediation of lead-related adverse renal effects. PMID- 16263505 TI - Within-home versus between-home variability of house dust endotoxin in a birth cohort. AB - Endotoxin exposure has been proposed as an environmental determinant of allergen responses in children. To better understand the implications of using a single measurement of house dust endotoxin to characterize exposure in the first year of life, we evaluated room-specific within-home and between-home variability in dust endotoxin obtained from 470 households in Boston, Massachusetts. Homes were sampled up to two times over 5-11 months. We analyzed 1,287 dust samples from the kitchen, family room, and baby's bedroom for endotoxin. We fit a mixed-effects model to estimate mean levels and the variation of endotoxin between homes, between rooms, and between sampling times. Endotoxin ranged from 2 to 1,945 units per milligram of dust. Levels were highest during summer and lowest in the winter. Mean endotoxin levels varied significantly from room to room. Cross sectionally, endotoxin was moderately correlated between family room and bedroom floor (r = 0.30), between family room and kitchen (r = 0.32), and between kitchen and bedroom (r = 0.42). Adjusting for season, the correlation of endotoxin levels within homes over time was 0.65 for both the bedroom and kitchen and 0.54 for the family room. The temporal within-home variance of endotoxin was lowest for bedroom floor samples and highest for kitchen samples. Between-home variance was lowest in the family room and highest for kitchen samples. Adjusting for season, within-home variation was less than between-home variation for all three rooms. These results suggest that room-to-room and home-to-home differences in endotoxin influence the total variability more than factors affecting endotoxin levels within a room over time. PMID- 16263506 TI - Some environmental contaminants influence motor and feeding behaviors in the ornate wrasse (Thalassoma pavo) via distinct cerebral histamine receptor subtypes. AB - Common environmental contaminants such as heavy metals and pesticides pose serious risks to behavioral and neuroendocrine functions of many aquatic organisms. In the present study, we show that the heavy metal cadmium and the pesticide endosulfan produce such effects through an interaction of specific cerebral histamine receptor subtypes in the teleost ornate wrasse (Thalassoma pavo). Treatment of this teleost with toxic cadmium levels for 1 week was sufficient to induce abnormal swimming movements, whereas reduced feeding behaviors were provoked predominantly by elevated endosulfan concentrations. In the brain, these environmental contaminants caused neuronal degeneration in cerebral targets such as the mesencephalon and hypothalamus, damage that appeared to correlate with altered binding levels of the three major histamine receptors (subtypes 1, 2, and 3). Although cadmium accounted for reduced binding activity of all three subtypes in most brain regions, it was subtype 2 that seemed to be its main target, as shown by a very great (p < 0.001) down-regulation in mesencephalic areas such as the stratum griseum central layer. Conversely, endosulfan provided very great and great (p < 0.01) up-regulating effects of subtype 3 and 1 levels, respectively, in preoptic-hypothalamic areas such as the medial part of the lateral tuberal nucleus, and in the suprachiasmatic nucleus. These results suggest that the neurotoxicant-dependent abnormal motor and feeding behaviors may well be tightly linked to binding activities of distinct histamine subtypes in localized brain regions of the Thalassoma pavo. PMID- 16263507 TI - Personal care product use predicts urinary concentrations of some phthalate monoesters. AB - Phthalates are multifunctional chemicals used in a variety of applications, including personal care products. The present study explored the relationship between patterns of personal care product use and urinary levels of several phthalate metabolites. Subjects include 406 men who participated in an ongoing semen quality study at the Massachusetts General Hospital Andrology Laboratory between January 2000 and February 2003. A nurse-administered questionnaire was used to determine use of personal care products, including cologne, aftershave, lotions, hair products, and deodorants. Phthalate monoester concentrations were measured in a single spot urine sample by isotope dilution-high-performance liquid chromatography coupled to tandem mass spectrometry. Men who used cologne or aftershave within 48 hr before urine collection had higher median levels of monoethyl phthalate (MEP) (265 and 266 ng/mL, respectively) than those who did not use cologne or aftershave (108 and 133 ng/mL, respectively). For each additional type of product used, MEP increased 33% (95% confidence interval, 14 53%). The use of lotion was associated with lower urinary levels of monobutyl phthalate (MBP) (14.9 ng/mL), monobenzyl phthalate (MBzP) (6.1 ng/mL), and mono(2 ethylhexyl) phthalate (MEHP) (4.4 ng/mL) compared with men who did not use lotion (MBP, 16.8 ng/mL; MBzP, 8.6 ng/mL; MEHP, 7.2 ng/mL). The identification of personal care products as contributors to phthalate body burden is an important step in exposure characterization. Further work in this area is needed to identify other predictors of phthalate exposure. PMID- 16263508 TI - Induction of proinflammatory cytokines and C-reactive protein in human macrophage cell line U937 exposed to air pollution particulates. AB - Exposure to particulate matter air pollution causes inflammatory responses and is associated with the progression of atherosclerosis and increased cardiovascular mortality. Macrophages play a key role in atherogenesis by releasing proinflammatory cytokines and forming foam cells in subendothelial lesions. The present study quantified the inflammatory response in a human macrophage cell line (U937) after exposure to an ambient particulate sample from urban dust (UDP) and a diesel exhaust particulate (DEP). The effect of native UDP and DEP was compared with their corresponding organic extracts (OE-UDP/OE-DEP) and stripped particles (sUDP/sDEP) to clarify their respective roles. Exposure to OE-UDP, OE DEP, UDP, DEP, and 2,3,7,8-tetrachlorodibenzo-p-dioxin led to a greater increase of interleukin (IL)-8, tumor necrosis factor-alpha, and cyclooxygenase-2 mRNA expression than did the stripped particles, whereas sUDP, sDEP, UDP, and DEP led to a greater production of C-reactive protein and IL-6 mRNA. The particles and the organic extract-induced expression of cyclooxygenase-2 and cytochrome P450 (CYP)1a1 was significantly suppressed by co-treatment with an aryl hydrocarbon receptor (AhR) antagonist, indicating that these effects are mainly mediated by the organic components, which can activate the AhR and CYP1a1. In contrast, the induction of C-reactive protein and IL-6 seems to be a particle-related effect that is AhR independent. The inflammatory response induced by particulate matter was associated with a subsequent increase of cholesterol accumulation, a hallmark of foam cells. Together, these data illustrate the interaction between particulate matter and the inflammatory response as well as the formation of cholesterol-accumulating foam cells, which are early markers of cardiovascular disease. PMID- 16263510 TI - Thyroid-hormone-disrupting chemicals: evidence for dose-dependent additivity or synergism. AB - Endocrine disruption from environmental contaminants has been linked to a broad spectrum of adverse outcomes. One concern about endocrine-disrupting xenobiotics is the potential for additive or synergistic (i.e., greater-than-additive) effects of mixtures. A short-term dosing model to examine the effects of environmental mixtures on thyroid homeostasis has been developed. Prototypic thyroid-disrupting chemicals (TDCs) such as dioxins, polychlorinated biphenyls (PCBs), and polybrominated diphenyl ethers have been shown to alter thyroid hormone homeostasis in this model primarily by up-regulating hepatic catabolism of thyroid hormones via at least two mechanisms. Our present effort tested the hypothesis that a mixture of TDCs will affect serum total thyroxine (T4) concentrations in a dose-additive manner. Young female Long-Evans rats were dosed via gavage with 18 different polyhalogenated aromatic hydrocarbons [2 dioxins, 4 dibenzofurans, and 12 PCBs, including dioxin-like and non-dioxin-like PCBs] for 4 consecutive days. Serum total T4 was measured via radioimmunoassay in samples collected 24 hr after the last dose. Extensive dose-response functions (based on seven to nine doses per chemical) were determined for individual chemicals. A mixture was custom synthesized with the ratio of chemicals based on environmental concentrations. Serial dilutions of this mixture ranged from approximately background levels to 100-fold greater than background human daily intakes. Six serial dilutions of the mixture were tested in the same 4-day assay. Doses of individual chemicals that were associated with a 30% TH decrease from control (ED30), as well as predicted mixture outcomes were calculated using a flexible single-chemical-required method applicable to chemicals with differing dose thresholds and maximum-effect asymptotes. The single-chemical data were modeled without and with the mixture data to determine, respectively, the expected mixture response (the additivity model) and the experimentally observed mixture response (the empirical model). A likelihood-ratio test revealed statistically significant departure from dose additivity. There was no deviation from additivity at the lowest doses of the mixture, but there was a greater-than additive effect at the three highest mixtures doses. At high doses the additivity model underpredicted the empirical effects by 2- to 3-fold. These are the first results to suggest dose-dependent additivity and synergism in TDCs that may act via different mechanisms in a complex mixture. The results imply that cumulative risk approaches be considered when assessing the risk of exposure to chemical mixtures that contain TDCs. PMID- 16263509 TI - Health effects of a mixture of indoor air volatile organics, their ozone oxidation products, and stress. AB - In our present study we tested the health effects among women of controlled exposures to volatile organic compounds (VOCs), with and without ozone (O3), and psychological stress. Each subject was exposed to the following three conditions at 1-week intervals (within-subject factor): VOCs (26 mg/m3), VOCs + O3 (26 mg/m3 + 40 ppb), and ambient air with a 1-min spike of VOCs (2.5 mg/m3). As a between subjects factor, half the subjects were randomly assigned to perform a stressor. Subjects were 130 healthy women (mean age, 27.2 years; mean education, 15.2 years). Health effects measured before, during, and after each 140-min exposure included symptoms, neurobehavioral performance, salivary cortisol, and lung function. Mixing VOCs with O3 was shown to produce irritating compounds including aldehydes, hydrogen peroxide, organic acids, secondary organic aerosols, and ultrafine particles (particulate matter with aerodynamic diameter < 0.1 microm). Exposure to VOCs with and without O3 did not result in significant subjective or objective health effects. Psychological stress significantly increased salivary cortisol and symptoms of anxiety regardless of exposure condition. Neither lung function nor neurobehavioral performance was compromised by exposure to VOCs or VOCs + O3. Although numerous epidemiologic studies suggest that symptoms are significantly increased among workers in buildings with poor ventilation and mixtures of VOCs, our acute exposure study was not consistent with these epidemiologic findings. Stress appears to be a more significant factor than chemical exposures in affecting some of the health end points measured in our present study. PMID- 16263511 TI - Ultrafine particles cross cellular membranes by nonphagocytic mechanisms in lungs and in cultured cells. AB - High concentrations of airborne particles have been associated with increased pulmonary and cardiovascular mortality, with indications of a specific toxicologic role for ultrafine particles (UFPs; particles < 0.1 microm). Within hours after the respiratory system is exposed to UFPs, the UFPs may appear in many compartments of the body, including the liver, heart, and nervous system. To date, the mechanisms by which UFPs penetrate boundary membranes and the distribution of UFPs within tissue compartments of their primary and secondary target organs are largely unknown. We combined different experimental approaches to study the distribution of UFPs in lungs and their uptake by cells. In the in vivo experiments, rats inhaled an ultrafine titanium dioxide aerosol of 22 nm count median diameter. The intrapulmonary distribution of particles was analyzed 1 hr or 24 hr after the end of exposure, using energy-filtering transmission electron microscopy for elemental microanalysis of individual particles. In an in vitro study, we exposed pulmonary macrophages and red blood cells to fluorescent polystyrene microspheres (1, 0.2, and 0.078 microm) and assessed particle uptake by confocal laser scanning microscopy. Inhaled ultrafine titanium dioxide particles were found on the luminal side of airways and alveoli, in all major lung tissue compartments and cells, and within capillaries. Particle uptake in vitro into cells did not occur by any of the expected endocytic processes, but rather by diffusion or adhesive interactions. Particles within cells are not membrane bound and hence have direct access to intracellular proteins, organelles, and DNA, which may greatly enhance their toxic potential. PMID- 16263512 TI - Consistent pulmonary and systemic responses from inhalation of fine concentrated ambient particles: roles of rat strains used and physicochemical properties. AB - Several studies have reported health effects of concentrated ambient particles (CAP) in rodents and humans; however, toxicity end points in rodents have provided inconsistent results. In 2000 we conducted six 1-day exposure studies where spontaneously hypertensive (SH) rats were exposed to filtered air or CAPs (< or = 2.5 microm, 1,138-1,765 microg/m3) for 4 hr (analyzed 1-3 hr afterward). In seven 2-day exposure studies in 2001, SH and Wistar Kyoto (WKY) rats were exposed to filtered air or CAP (< or = 2.5 microm, 144-2,758 microg/m3) for 4 hr/day times 2 days (analyzed 1 day afterward). Despite consistent and high CAP concentrations in the 1-day exposure studies, no biologic effects were noted. The exposure concentrations varied among the seven 2-day exposure studies. Except in the first study when CAP concentration was highest, lavageable total cells and macrophages decreased and neutrophils increased in WKY rats. SH rats demonstrated a consistent increase of lavage fluid gamma-glutamyltransferase activity and plasma fibrinogen. Inspiratory and expiratory times increased in SH but not in WKY rats. Significant correlations were found between CAP mass (microgram per cubic meter) and sulfate, organic carbon, or zinc. No biologic effects correlated with CAP mass. Despite low chamber mass in the last six of seven 2-day exposure studies, the levels of zinc, copper, and aluminum were enriched severalfold, and organic carbon was increased to some extent when expressed per milligram of CAP. Biologic effects were evident in those six studies. These studies demonstrate a pattern of rat strain-specific pulmonary and systemic effects that are not linked to high mass but appear to be dependent on CAP chemical composition. PMID- 16263513 TI - Do organohalogen contaminants contribute to histopathology in liver from East Greenland polar bears (Ursus maritimus)? AB - In East Greenland polar bears (Ursus maritimus), anthropogenic organohalogen compounds (OHCs) (e.g., polychlorinated biphenyls, dichlorodiphenyltrichloroethane, and polybrominated diphenyl ethers) contributed to renal lesions and are believed to reduce bone mineral density. Because OHCs are also hepatotoxic, we investigated liver histology of 32 subadult, 24 adult female, and 23 adult male East Greenland polar bears sampled during 1999-2002. Light microscopic changes consisted of nuclear displacement from the normal central cytoplasmic location in parenchymal cells, mononuclear cell infiltrations (mainly portally and as lipid granulomas), mild bile duct proliferation accompanied by fibrosis, and fat accumulation in hepatocytes and pluripotent Ito cells. Lipid accumulation in Ito cells and bile duct hyperplasia accompanied by portal fibrosis were correlated to age, whereas no changes were associated with either sex or season (summer vs. winter). For adult females, hepatocytic intracellular fat increased significantly with concentrations of the sum of hexachlorocyclohexanes, as was the case for lipid granulomas and hexachlorobenzene in adult males. Based on these relationships and the nature of the chronic inflammation, we suggest that these findings were caused by aging and long-term exposure to OHCs. Therefore, these changes may be used as biomarkers for OHC exposure in wildlife and humans. To our knowledge, this is the first time liver histology has been evaluated in relation to OHC concentrations in a mammalian wildlife species, and the information is important to future polar bear conservation strategies and health assessments of humans relying on OHC contaminated food resources. PMID- 16263514 TI - PM source apportionment for short-term cardiac function changes in ApoE-/- mice. AB - Daily rates of cardiovascular mortality and morbidity are have been associated with daily variations in fine particulate matter (aerodynamic diameter < or = 2.5 microm, PM2.5), but little is known about the influences of the individual source related PM2.5 categories or the temporal lags for the effects. We investigated heart rate (HR) and HR variability (HRV) data collected during a 5-month study involving 6 hr/day, 5 day/week exposures of normal (C57) mice and a murine model for atherosclerotic disease (ApoE-/-) in Sterling Forest (Tuxedo, New York, USA). The mice were exposed to concentrated ambient particles (PM2.5 concentrated 10 fold, producing an average of 113 microg/m3). Daily 6-hr PM2.5 air samples were analyzed by X-ray fluorescence, permitting attribution to major PM source categories [secondary sulfate (SS), resuspended soil (RS), residual oil (RO) combustion, and other, largely due to motor vehicle traffic]. We examined associations between these PM2.5 components and both HR and HRV for three different daily time periods: during exposure, the afternoon after exposure, and late at night. For HR there were significant transient associations for RS during exposure, and for SS in the afternoon after exposure. For HRV, there were comparable associations with RO in the afternoon after exposure and for both SS and RS late at night. The biologic bases for these associations and their temporal lags are not known but may be related to the differential solubility of the biologically active PM components at the respiratory epithelia and their access to cells that release mediators that reach the cardiovascular system. Clearly, further research to elucidate the underlying processes is needed. PMID- 16263515 TI - Cellular and hormonal disruption of fetal testis development in sheep reared on pasture treated with sewage sludge. AB - The purpose of this study was to evaluate whether experimental exposure of pregnant sheep to a mixture of environmental chemicals added to pasture as sewage sludge (n = 9 treated animals) exerted effects on fetal testis development or function; application of sewage sludge was undertaken so as to maximize exposure of the ewes to its contents. Control ewes (n = 9) were reared on pasture treated with an equivalent amount of inorganic nitrogenous fertilizer. Treatment had no effect on body weight of ewes, but it reduced body weight by 12-15% in male (n = 12) and female (n = 8) fetuses on gestation day 110. In treated male fetuses (n = 11), testis weight was significantly reduced (32%), as were the numbers of Sertoli cells (34% reduction), Leydig cells (37% reduction), and gonocytes (44% reduction), compared with control fetuses (n = 8). Fetal blood levels of testosterone and inhibin A were also reduced (36% and 38%, respectively) in treated compared with control fetuses, whereas blood levels of luteinizing hormone and follicle-stimulating hormone were unchanged. Based on immunoexpression of anti-Mullerian hormone, cytochrome P450 side chain cleavage enzyme, and Leydig cell cytoplasmic volume, we conclude that the hormone changes in treated male fetuses probably result from the reduction in somatic cell numbers. This reduction could result from fetal growth restriction in male fetuses and/or from the lowered testosterone action; reduced immunoexpression of alpha-smooth muscle actin in peritubular cells and of androgen receptor in testes of treated animals supports the latter possibility. These findings indicate that exposure of the developing male sheep fetus to real-world mixtures of environmental chemicals can result in major attenuation of testicular development and hormonal function, which may have consequences in adulthood. PMID- 16263516 TI - A rapid, physiologic protocol for testing transcriptional effects of thyroid disrupting agents in premetamorphic Xenopus tadpoles. AB - Increasing numbers of substances present in the environment are postulated to have endocrine-disrupting effects on vertebrate populations. However, data on disruption of thyroid signaling are fragmentary, particularly at the molecular level. Thyroid hormone (TH; triiodothyronine, T3) acts principally by modulating transcription from target genes; thus, thyroid signaling is particularly amenable to analysis with a transcriptional assay. Also, T3 orchestrates amphibian metamorphosis, thereby providing an exceptional model for identifying thyroid disrupting chemicals. We combined these two advantages to develop a method for following and quantifying the transcriptional action of T3 in Xenopus laevis tadpoles. This technology provides a means of assessing thyroid activity at the molecular level in a physiologically relevant situation. Moreover, translucent tadpoles are amenable to "on-line" imaging with fluorescent reporter constructs that facilitate in vivo measurement of transcriptional activity. We adapted transgenesis with TH-responsive elements coupled to either luciferase or green fluorescent protein to follow T3-dependent transcription in vivo. To reduce time of exposure and to synchronize responses, we optimized a physiologic pretreatment protocol that induced competence to respond to T3 and thus to assess T3 effects and T3 disruption within 48 hr. This pretreatment protocol was based on a short (24 hr), weak (10(-12) M) pulse of T3 that induced TH receptors, facilitating and synchronizing the transcriptional responses. This protocol was successfully applied to somatic and germinal transgenesis with both reporter systems. Finally, we show that the transcriptional assay allows detection of the thyroid-disrupting activity of environmentally relevant concentrations (10(-8) M) of acetochlor, a persistent herbicide. PMID- 16263517 TI - Systemic effects of arctic pollutants in beluga whales indicated by CYP1A1 expression. AB - Cytochrome P450 1A1 (CYP1A1) is induced by exposure to polycyclic aromatic hydrocarbons (PAHs) and planar halogenated aromatic hydrocarbons (PHAHs) such as non-ortho polychlorinated biphenyls (PCBs). In this study, we examined CYP1A1 protein expression immunohistochemically in multiple organs of beluga whales from two locations in the Arctic and from the St. Lawrence estuary. These beluga populations have some of the lowest (Arctic sites) and highest (St. Lawrence estuary) concentrations of PCBs in blubber of all cetaceans. Samples from these populations might be expected to have different contaminant-induced responses, reflecting their different exposure histories. The pattern and extent of CYP1A1 staining in whales from all three locations were similar to those seen in animal models in which CYP1A has been highly induced, indicating a high-level expression in these whales. CYP1A1 induction has been related to toxic effects of PHAHs or PAHs in some species. In St. Lawrence beluga, the high level of CYP1A1 expression coupled with high levels of contaminants (including CYP1A1 substrates, e.g., PAH procarcinogens potentially activated by CYP1A1) indicates that CYP1A1 could be involved in the development of neoplastic lesions seen in the St. Lawrence beluga population. The systemic high-level expression of CYP1A1 in Arctic beluga suggests that effects of PAHs or PHAHs may be expected in Arctic populations, as well. The high-level expression of CYP1A1 in the Arctic beluga suggests that this species is highly sensitive to CYP1A1 induction by aryl hydrocarbon receptor agonists. PMID- 16263518 TI - Organotins disrupt the 11beta-hydroxysteroid dehydrogenase type 2-dependent local inactivation of glucocorticoids. AB - Organotins, important environmental pollutants widely used in agricultural and industrial applications, accumulate in the food chain and induce imposex in several marine species as well as neurotoxic and immunotoxic effects in higher animals. Reduced birth weight and thymus involution, observed upon exposure to organotins, can also be caused by excessive glucocorticoid levels. We now demonstrate that organotins efficiently inhibit 11beta-hydroxysteroid dehydrogenase type 2 (11beta-HSD2), converting active 11beta hydroxyglucocorticoids into inactive 11-ketoglucocorticoids, but not 11beta-HSD1, which catalyzes the reverse reaction. Di- and tributyltin as well as di- and triphenyltin inhibited recombinant and endogenous 11beta-HSD2 in lysates and intact cells with IC50 values between 500 nM and 3 microM. Dithiothreitol protected 11beta-HSD2 from organotin-dependent inhibition, indicating that organotins act by binding to one or more cysteines. Mutational analysis and 3-D structural modeling revealed several important interactions of cysteines in 11beta-HSD2. Cys90, Cys228, and Cys264 were essential for enzymatic stability and catalytic activity, suggesting that disruption of such interactions by organotins leads to inhibition of 11beta-HSD2. Enhanced glucocorticoid concentrations due to disruption of 11beta-HSD2 function may contribute to the observed organotin dependent toxicity in some glucocorticoid-sensitive tissues such as thymus and placenta. PMID- 16263520 TI - Workgroup report: Biomonitoring study design, interpretation, and communication- lessons learned and path forward. AB - Human biomonitoring investigations have provided data on a wide array of chemicals in blood and urine and in other tissues and fluids such as hair and human milk. These data have prompted questions such as a) What is the relationship between levels of environmental chemicals in humans and external exposures? b) What is the baseline or "background" level against which individual levels should be compared? and c) How can internal levels be used to draw conclusions about individual and/or population health? An interdisciplinary panel was convened for a 1-day workshop in November 2004 with the charge of focusing on three specific aspects of biomonitoring: characteristics of scientifically robust biomonitoring studies, interpretation of human biomonitoring data for potential risks to human health, and communication of results, uncertainties, and limitations of biomonitoring studies. In this report we describe the recommendations of the panel. PMID- 16263519 TI - Workgroup report: Drinking-water nitrate and health--recent findings and research needs. AB - Human alteration of the nitrogen cycle has resulted in steadily accumulating nitrate in our water resources. The U.S. maximum contaminant level and World Health Organization guidelines for nitrate in drinking water were promulgated to protect infants from developing methemoglobinemia, an acute condition. Some scientists have recently suggested that the regulatory limit for nitrate is overly conservative; however, they have not thoroughly considered chronic health outcomes. In August 2004, a symposium on drinking-water nitrate and health was held at the International Society for Environmental Epidemiology meeting to evaluate nitrate exposures and associated health effects in relation to the current regulatory limit. The contribution of drinking-water nitrate toward endogenous formation of N-nitroso compounds was evaluated with a focus toward identifying subpopulations with increased rates of nitrosation. Adverse health effects may be the result of a complex interaction of the amount of nitrate ingested, the concomitant ingestion of nitrosation cofactors and precursors, and specific medical conditions that increase nitrosation. Workshop participants concluded that more experimental studies are needed and that a particularly fruitful approach may be to conduct epidemiologic studies among susceptible subgroups with increased endogenous nitrosation. The few epidemiologic studies that have evaluated intake of nitrosation precursors and/or nitrosation inhibitors have observed elevated risks for colon cancer and neural tube defects associated with drinking-water nitrate concentrations below the regulatory limit. The role of drinking-water nitrate exposure as a risk factor for specific cancers, reproductive outcomes, and other chronic health effects must be studied more thoroughly before changes to the regulatory level for nitrate in drinking water can be considered. PMID- 16263521 TI - Meeting report: Structural determination of environmentally responsive proteins. AB - The three-dimensional structure of gene products continues to be a missing lynchpin between linear genome sequences and our understanding of the normal and abnormal function of proteins and pathways. Enhanced activity in this area is likely to lead to better understanding of how discrete changes in molecular patterns and conformation underlie functional changes in protein complexes and, with it, sensitivity of an individual to an exposure. The National Institute of Environmental Health Sciences convened a workshop of experts in structural determination and environmental health to solicit advice for future research in structural resolution relative to environmentally responsive proteins and pathways. The highest priorities recommended by the workshop were to support studies of structure, analysis, control, and design of conformational and functional states at molecular resolution for environmentally responsive molecules and complexes; promote understanding of dynamics, kinetics, and ligand responses; investigate the mechanisms and steps in posttranslational modifications, protein partnering, impact of genetic polymorphisms on structure/function, and ligand interactions; and encourage integrated experimental and computational approaches. The workshop participants also saw value in improving the throughput and purity of protein samples and macromolecular assemblies; developing optimal processes for design, production, and assembly of macromolecular complexes; encouraging studies on protein-protein and macromolecular interactions; and examining assemblies of individual proteins and their functions in pathways of interest for environmental health. PMID- 16263522 TI - Tubular and glomerular kidney effects in Swedish women with low environmental cadmium exposure. AB - Cadmium is a well-known nephrotoxic agent in food and tobacco, but the exposure level that is critical for kidney effects in the general population is not defined. Within a population-based women's health survey in southern Sweden (Women's Health in the Lund Area, WHILA), we investigated cadmium exposure in relation to tubular and glomerular function, from 1999 through early 2000 in 820 women (71% participation rate) 53-64 years of age. Multiple linear regression showed cadmium in blood (median, 0.38 microg/L) and urine (0.52 microg/L; density adjusted = 0.67 microg/g creatinine) to be significantly associated with effects on renal tubules (as indicated by increased levels of human complex-forming protein and N-acetyl-beta-D-glucosaminidase in urine), after adjusting for age, body mass index, blood lead, diabetes, hypertension, and regular use of nephrotoxic drugs. The associations remained significant even at the low exposure in women who had never smoked. We also found associations with markers of glomerular effects: glomerular filtration rate and creatinine clearance. Significant effects were seen already at a mean urinary cadmium level of 0.6 microg/L (0.8 microg/g creatinine). Cadmium potentiated diabetes-induced effects on kidney. In conclusion, tubular renal effects occurred at lower cadmium levels than previously demonstrated, and more important, glomerular effects were also observed. Although the effects were small, they may represent early signs of adverse effects, affecting large segments of the population. Subjects with diabetes seem to be at increased risk. PMID- 16263523 TI - Decline of ambient air pollution levels and improved respiratory health in Swiss children. AB - The causality of observed associations between air pollution and respiratory health in children is still subject to debate. If reduced air pollution exposure resulted in improved respiratory health of children, this would argue in favor of a causal relation. We investigated whether a rather moderate decline of air pollution levels in the 1990s in Switzerland was associated with a reduction in respiratory symptoms and diseases in school children. In nine Swiss communities, 9,591 children participated in cross-sectional health assessments between 1992 and 2001. Their parents completed identical questionnaires on health status and covariates. We assigned to each child an estimate of regional particles with an aerodynamic diameter < 10 microg/m3 (PM10) and determined change in PM10 since the first survey. Adjusted for socioeconomic, health-related, and indoor factors, declining PM10 was associated in logistic regression models with declining prevalence of chronic cough [odds ratio (OR) per 10-microg/m3 decline = 0.65, 95% confidence interval (CI), 0.54-0.79], bronchitis (OR = 0.66; 95% CI, 0.55-0.80), common cold (OR = 0.78; 95% CI, 0.68-0.89), nocturnal dry cough (OR = 0.70; 95% CI, 0.60-0.83), and conjunctivitis symptoms (OR = 0.81; 95% CI, 0.70-0.95). Changes in prevalence of sneezing during pollen season, asthma, and hay fever were not associated with the PM10 reduction. Our findings show that the reduction of air pollution exposures contributes to improved respiratory health in children. No threshold of adverse effects of PM10 was apparent because we observed the beneficial effects for relatively small changes of rather moderate air pollution levels. Current air pollution levels in Switzerland still exceed limit values of the Swiss Clean Air Act; thus, children's health can be improved further. PMID- 16263524 TI - Birth outcomes and prenatal exposure to ozone, carbon monoxide, and particulate matter: results from the Children's Health Study. AB - Exposures to ambient air pollutants have been associated with adverse birth outcomes. We investigated the effects of air pollutants on birth weight mediated by reduced fetal growth among term infants who were born in California during 1975-1987 and who participated in the Children's Health Study. Birth certificates provided maternal reproductive history and residence location at birth. Sociodemographic factors and maternal smoking during pregnancy were collected by questionnaire. Monthly average air pollutant levels were interpolated from monitors to the ZIP code of maternal residence at childbirth. Results from linear mixed-effects regression models showed that a 12-ppb increase in 24-hr ozone averaged over the entire pregnancy was associated with 47.2 g lower birth weight [95% confidence interval (CI), 27.4-67.0 g], and this association was most robust for exposures during the second and third trimesters. A 1.4-ppm difference in first-trimester carbon monoxide exposure was associated with 21.7 g lower birth weight (95% CI, 1.1-42.3 g) and 20% increased risk of intrauterine growth retardation (95% CI, 1.0-1.4). First-trimester CO and third-trimester O3 exposures were associated with 20% increased risk of intrauterine growth retardation. A 20-microg/m3 difference in levels of particulate matter < or = 10 microm in aerodynamic diameter (PM10) during the third trimester was associated with a 21.7-g lower birth weight (95% CI, 1.1-42.2 g), but this association was reduced and not significant after adjusting for O3. In summary, O3 exposure during the second and third trimesters and CO exposure during the first trimester were associated with reduced birth weight. PMID- 16263525 TI - In utero exposure to dioxins and polychlorinated biphenyls and its relations to thyroid function and growth hormone in newborns. AB - The aim of this study is to examine the association between transplacental exposure to dioxins/polychlorinated biphenyls (PCBs) and thyroid and growth hormones in newborns. We recruited 118 pregnant women, between 25 and 34 years of age, at the obstetric clinic. Personal data collected included reproductive and medical histories and physical factors. Clinicians gathered placental and umbilical cord serum upon delivery and carefully scored the 118 newborns, making both structural and functional assessments. We analyzed placentas for 17 polychlorinated dibenzo-p-dioxins and dibenzofurans and 12 dioxin-like PCB congeners with the World Health Organization-defined toxic equivalent factors, and six indicator PCBs by high-resolution gas chromatography and high-resolution mass spectrometry. We analyzed thyroid and growth hormones from cord serum using radioimmunoassay. Insulin-like growth factor (IGF)-1, IGF-binding globulin-3, and thyroxine x thyroid-stimulating hormone (T4 x TSH) were significantly associated with increased placental weight and Quetelet index (in kilograms per square meter; correlation coefficient r = 0.2-0.3; p < 0.05). Multivariate analyses showed independently and significantly decreased free T4 (FT4) x TSH with increasing non-ortho PCBs (r = -0.2; p < 0.05). We suggest that significant FT4 feedback alterations to the hypothalamus result from in utero exposure to non ortho PCBs. Considering the vast existence of bioaccumulated dioxins and PCBs and the resultant body burden in modern society, we suggest routine screening of both thyroid hormone levels and thyroid function in newborns. PMID- 16263527 TI - Contraceptive use among the poor in Indonesia. AB - CONTEXT: Indonesia has experienced a dramatic increase in contraceptive use and an equally dramatic fertility decline over the last 30 years. Yet recent reductions in family planning funding, program decentralization and the diminishing role of the public sector as a service provider may lead to lower use among poor women. METHODS: The data for analysis were drawn from the 2002-2003 Indonesia Demographic and Health Survey. Bivariate techniques were used to compare overall contraceptive use, reliance on specific methods, source of supplies and reasons for nonuse of contraceptives between poor and better-off women. Multivariate regression assessed the association between use of a modern method and selected social, demographic and attitudinal characteristics. RESULTS: Better-off women wanted significantly fewer children than did moderately or extremely poor women (2.8 vs. 3.0-3.4), were more likely to approve of family planning (93% vs. 87-91%) and were more likely to believe their spouses approved (91% vs. 80-87%). Better-off women and moderately poor women had higher odds of using modern contraceptives than did extremely poor women (odds ratios, 1.6 and 1.4, respectively). Compared with women who gave a non-numeric response, those who wanted two or fewer children had higher odds of using a modern method (2.0). The odds were also higher among women who lived in a district in which the mean ideal number of children was below the national median (1.5). CONCLUSION: Governmental efforts to increase contraceptive use among poor women need to focus on changing attitudes toward smaller family sizes and family planning. PMID- 16263526 TI - Biologic monitoring to characterize organophosphorus pesticide exposure among children and workers: an analysis of recent studies in Washington State. AB - We examined findings from five organophosphorus pesticide biomonitoring studies conducted in Washington State between 1994 and 1999. We compared urinary dimethylthiophosphate (DMTP) concentrations for all study groups and composite dimethyl alkylphosphate (DMAP) concentrations for selected groups. Children of pesticide applicators had substantially higher metabolite levels than did Seattle children and farmworker children (median DMTP, 25 microg/L; p < 0.0001). Metabolite levels of children living in agricultural communities were elevated during periods of crop spraying. Median DMTP concentrations for Seattle children and farmworker children did not differ significantly (6.1 and 5.8 microg/L DMTP, respectively; p = 0.73); however, the DMAP concentrations were higher for Seattle children than for farmworker children (117 and 87 nmol/L DMAP, respectively; p = 0.007). DMTP concentrations of U.S. children 6-11 years of age (1999-2000 National Health and Nutrition Examination Survey population) were higher than those of Seattle children and farmworker children at the 75th, 90th, and 95th percentiles. DMTP concentrations for workers actively engaged in apple thinning were 50 times higher than DMTP concentrations for farmworkers sampled outside of peak exposure periods. We conclude that workers who have direct contact with pesticides should continue to be the focus of public health interventions and that elevated child exposures in agricultural communities may occur during active crop-spraying periods and from living with a pesticide applicator. Timing of sample collection is critical for the proper interpretation of pesticide biomarkers excreted relatively soon after exposure. We surmise that differences in dietary exposure can explain the similar exposures observed among farmworker children, children living in the Seattle metropolitan area, and children sampled nationally. PMID- 16263528 TI - Use of family planning services in the transition to a static clinic system in Bangladesh: 1998-2002. AB - CONTEXT: In rural Bangladesh, family planning services--previously provided through household visits and satellite clinics--were transferred to static community clinics under the government's sectoral program for 1998-2003, but the next sectoral program reversed the change without a formal evaluation. It is important to assess changes in utilization and coverage to inform further development of the service delivery system. METHODS: Longitudinal data on use of family planning services and contraceptive methods were collected quarterly in 1998-2002 from married women in about 11,000 households in two rural surveillance areas--Abhoynagar and Mirsarai. Cross-sectional surveys were conducted among women and service providers in 2003 to gather detailed information about the transition to static clinics and women's response to the changes. Quarterly time series graphs of selected indicators were plotted for areas served by community clinics. RESULTS: In a time of considerable change in service delivery and sources of contraceptive supply, contraceptive prevalence remained constant in Abhoynagar and increased in Mirsarai. Community clinics quickly became the source of supplies for one-third of contraceptive users in Abhoynagar and one-fifth in Mirsarai. In wards where community clinics became operational (mostly in 2001 2002), three-quarters of women had used one at some time. CONCLUSIONS: Despite cultural constraints on mobility, women do not appear to have become dependent on home delivery of contraceptives. PMID- 16263529 TI - Gender inequality and intimate partner violence among women in Moshi, Tanzania. AB - CONTEXT: In Sub-Saharan Africa, where rates of intimate partner violence are high, knowing the prevalence of abuse and associated patterns of risk is crucial to ensuring women's health and development. Intimate partner violence in Tanzania has not been assessed through a population-based survey. METHODS: A household based sample of women aged 20-44 in the urban district of Moshi, Tanzania, participated in face-to-face interviews in 2002-2003. The lifetime prevalence of exposure to intimate partner violence and the prevalence of exposure during the past 12 months were assessed among 1,444 women who reported having a current partner. Multivariate logistic regression was used to identify factors associated with intimate partner violence. RESULTS: Twenty-one percent of women reported having experienced intimate partner violence (i.e., having been threatened with physical abuse, subjected to physical abuse or forced into intercourse by a partner) during the previous 12 months; 26% reported such an experience at any time, including the past 12 months. The likelihood of violence in the past year was elevated if the woman had had problems conceiving or had borne five or more children (odds ratios, 1.9 and 2.4, respectively); if her husband or partner had other partners (2.0) or contributed little to expenses for her and her children (3.3); and if she had had no more than a primary education (1.7). CONCLUSIONS: Gender inequality within sexual unions is associated with intimate partner violence. Policies and programs that discourage men from blaming women for infertility, promote monogamous unions and expand access to education for women may reduce intimate partner violence in northern urban Tanzania. PMID- 16263530 TI - Gender relations and reproductive decision making in Honduras. AB - CONTEXT: Gender differences influence decision making about reproductive health. Most information on reproductive health decision making in Latin America has come from women's reports of men's involvement. METHODS: Data were collected in Honduras in 2001 through two national surveys that used independent samples of men aged 15-59 years and women aged 15-49. Bivariate and multivariate analyses were used to identify factors associated with male-centered decision-making attitudes and behaviors regarding family size and family planning use. RESULTS: Overall, 25% of women and 28% of men said that men alone should be responsible for at least one of these reproductive decisions, and 27% of women and 21% of men said that the man in their household made one or both decisions. For women, having no children and being in a consensual union were each associated with holding male-centered decision-making attitudes; having less than a secondary education, being of medium or low socioeconomic status and living in a rural area were each associated with male-centered decision making. Among men, having less than secondary education and being in a consensual union were each associated with male-centered decision-making attitudes and behavior. Women who had ever used or were currently using modern methods were significantly less likely to hold attitudes supporting male-centered decision-making than were those who relied on traditional methods and those who had never used a modern method. CONCLUSIONS: Programs should recognize power imbalances between genders that affect women's ability to meet their stated fertility desires. In rural areas, programs should target men, encouraging them to communicate with their wives on reproductive decisions. PMID- 16263531 TI - The incidence of induced abortion in the Philippines: current level and recent trends. AB - CONTEXT: In the Philippines, abortion is legally restricted. Nevertheless, many women obtain abortions--often in unsafe conditions--to avoid unplanned births. In 1994, the estimated abortion rate was 25 per 1,000 women per year; no further research on abortion incidence has been conducted in the Philippines. METHODS: Data from 1,658 hospitals were used to estimate abortion incidence in 2000 and to assess trends between 1994 and 2000, nationally and by region. An indirect estimation methodology was used to calculate the total number of women hospitalized for complications of induced abortion in 2000 (averaged data for 1999-2001), the total number of women having abortions and the rate of induced abortion. RESULTS: In 2000, an estimated 78,900 women were hospitalized for postabortion care, 473,400 women had abortions and the abortion rate was 27 per 1,000 women aged 15-44 per year. The national abortion rate changed little between 1994 and 2000; however, large increases occurred in metropolitan Manila (from 41 to 52) and Visayas (from 11 to 17). The proportions of unplanned births and unintended pregnancies increased substantially in Manila, and the use of traditional contraceptive methods increased in Manila and Visayas. CONCLUSION: The increase in the level of induced abortion seen in some areas may reflect the difficulties women experience in obtaining modern contraceptives as a result of social and political constraints that affect health care provision. Policies and programs regarding both postabortion care and contraceptive services need improvement. PMID- 16263532 TI - Frozen section: Its role in gynaecological oncology. AB - The use of frozen section has greatly impacted on the care of the gynaecological oncology patient. Frozen section allows intraoperative evaluation to distinguish benign from malignant tumors in order to tailor the extent of surgery necessary. Frozen section diagnosis in gynaecological oncology is sufficiently sensitive and specific for clinical use. Generally, the false negative rate is low and the false positive rate is negligible. Deferred diagnoses or incompatible frozen section diagnosis is usually due to technical limitations especially for the mucinous ovarian tumors. This review summarises the available literature on the accuracy, limitations and role of frozen section for individual gynaecological tumors. PMID- 16263533 TI - Risk communication: illusion or reality? PMID- 16263534 TI - Registering in a health facility for delivery protects against maternal mortality in a developing country setting. AB - A prospective study on the maternal deaths in public and private health facilities in Ogun State was undertaken for a period of 9 months (November 2003 - July 2004). While data were collected from all of the primary, the secondary and tertiary health facilities, it could only be collected from 123 (18.6%) of the private health facilities in the state. Of the 20,831 live births during the period, 37 maternal deaths occurred giving a maternal mortality ratio of 177.6 deaths per 100,000 live births. Some 84% of maternal deaths were not registered for delivery in the health facilities they delivered. The maternal mortality ratio is comparable with previously reported data for South-West Nigeria. A total of 81% of the deaths were due to three main causes: haemorrhage, eclampsia and infection. The maternal mortality ratio was almost three-fold greater in women above the age of 35 years. There is therefore a need to enhance activities to reduce maternal mortality. Private health facilities registration must be tied to sending data to authorized government departments. There is also a need to capture childbirth and attendant complications and death occurring outside orthodox health facilities. PMID- 16263535 TI - Breech presentation: an audit project as means of pursuing clinical excellence. AB - Clinical audit is an effective quality improvement process to evaluate important clinical issues. Breech presentation is such an issue due to its contribution to the rising caesarean section (CS) rate. We set out to assess the management of breech presentation using, as standards, the delivery suite protocol and national guidelines. Our first audit revealed a low success rate of external cephalic version (ECV) and deficient documentation of written consent for ECV, other aspects of care being satisfactory. The results were presented to a multidisciplinary meeting and disseminated to relevant stakeholders. A re-audit was then performed. It confirmed significant improvement in the documentation of consent for ECV. It also revealed a good detection rate of breech, optimal offer rate of ECV and good neonatal outcome. However, uptake of ECV as well as the success rate could both be improved so as to reduce the CS rate for breech presentation. We discuss options for improving the uptake and success rate for ECV. PMID- 16263536 TI - What investigation is appropriate following maternal perception of reduced fetal movements? AB - It has been proposed that maternal perception of reduced fetal movements may be indicative of placental insufficiency, and that women who present with reduced fetal movements should be investigated for evidence of placental insufficiency. A retrospective case notes-based analysis of patients presenting with reduced fetal movements was undertaken. There were 92 live-born infants, the mean gestation at delivery was 39(+4) weeks. The median birth weight was 3,140 g. A total of 29.1% of infants had a birth weight of less than the 10th centile. A total of 35 women underwent ultrasound assessment of fetal growth and liquor volume. Measurement of symphyseal fundal height (SFH) had a greater specificity than a single ultrasound assessment in the prediction of IUGR at delivery. To reduce unnecessary intervention, SFH measurement may be used as a screening tool for those who merit ultrasound assessment of growth and umbilical artery Doppler. PMID- 16263537 TI - Evaluation of the role of day assessment unit in the management of pregnancy induced hypertension. AB - This study is based on an audit designed to investigate observance of the guideline on pregnancy induced hypertension (PIH) and referral to the day assessment unit (DAU) at the Royal Berkshire Hospital. The audit included 60 patients; all were first time referrals, over a period of 4 months in 2003. The main sources of cases were the community (50%), followed by the antenatal clinic (ANC) 47%. The majority (67%) were referred with hypertension. A total of 28% had suspected symptomatic pre-eclampsia. About 94% of the patients were at >or= 32 weeks' gestation. After assessment in the unit, it was found that about 47% had a diastolic blood pressure of <90 mmHg and 77% had no proteinuria. Nearly all the cases referred with symptoms were found to be asymptomatic on review. The laboratory investigations were normal in over 92% of cases. In spite of these findings, 65% of patients were followed-up in ANC and DAU. The audit showed the need for the local guideline. PMID- 16263538 TI - Effectiveness of referral system for antenatal and intra-partum problems in Gutu district, Zimbabwe. AB - We conducted a population-based cohort study to determine the prevalence of antenatal and intra-partum referrals, compliance with advice and perinatal outcomes in referred pregnant women in Gutu district, Zimbabwe. The cohort was composed of 10,572 women who received antenatal care in 23 rural health centres (RHC) in Gutu district between January 1995 and June 1998. Pregnancy records of women with antenatal or intra-partum referral were analysed for indication, compliance and perinatal outcomes. Using women who had no antenatal referral or those who complied as referents, the association of referral with perinatal outcome was expressed as relative risk (RR) with 95% confidence intervals (CI). A total of 30% of women (3,094/10,572) had an antenatal referral. Among women attending RHC in labour, 13% (694/5,338) were referred intra-partum. Nulliparous and women younger than 20 years were more likely to be referred. Nurse - midwives' compliance with referral recommendations was low as 59% women with historical risk factors and 52% with raised blood pressure (>140/90 mmHg) were not referred. Women complied with referral advice except when indication was high parity. Women with antenatal referral were more likely to have hospital delivery, 70% vs 18% (p < 0.001). A total of 13% (993/7,478) of women referred themselves for hospital delivery. The risk of perinatal death was elevated among intra partum referrals (RR 3.4; 95% CI 1.7 - 6.8), self-referrals (RR 2.6; 95% CI 1.5 - 4.5) and also among women with historical risk factors who were not referred (RR 4.8; 95% CI 2.5 - 9.2). We concluded that although there was a functional referral system in Gutu district its efficiency was reduced by failure of health personnel to comply with referral recommendations. Women took appropriate action for most referral indications. PMID- 16263539 TI - Induction of labour after a previous caesarean section: a retrospective study in a district general hospital. AB - A retrospective study was undertaken in a district general hospital to identify factors associated with vaginal delivery, as opposed to caesarean section, in women undergoing induction of labour after a previous caesarean section. The study was undertaken over 9 years (April 1994 - May 2003) and included patients in their second or subsequent pregnancy who had previously had one lower segment caesarean delivery and in whom labour had been induced. Records were extracted from a database and anonymised. Vaginal delivery after induction of labour was attempted in 81 patients of whom 64 (79.0%) subsequently delivered vaginally. There were few complications and no cases of uterine rupture. Two factors had a statistical significant relationship with vaginal birth after induction of labour; occipito-anterior position (OR 10.18, 95% CI 1.42 - 112.7, Yates corrected chi2; p = 0.001) and more than one previous birth (OR 4.76, 95% CI 1.28 - 21.67, p = 0.017). Other associations were explored but were not statistically significant. This paper contributes to the literature on factors associated with vaginal delivery after induction of labour and previous caesarean section, which may inform the selection of cases, and consequent success rates for vaginal delivery. PMID- 16263540 TI - A prospective observational study of emergency caesarean section rates and the effect of the labour ward experience. AB - The National Sentinel Audit found that one in five births in England and Wales was by caesarean section. The reasons for the increase in caesarean section rates are multi-factorial. Anecdotally, it is suggested that obstetric intervention rates and caesarean section rates vary among obstetricians without a difference in fetal and maternal outcomes. The aim of this prospective observational study of 817 deliveries was to assess the effect of experience on the caesarean section rates for different obstetricians. Obstetricians with greater than 3 years of 2nd on-call labour ward experience had a statistically significant lower caesarean rate than less experienced obstetricians 10.25% vs 25.49%, respectively (p < 0.05). Differences in instrument deliveries was also studied. PMID- 16263542 TI - Adverse events in gynaecology at King Edward VIII Hospital, Durban, South Africa. AB - Audit of the quality of gynaecological care is essential so that errors due to medical interventions can be reduced to the bare minimum. The aim of this study was to determine the incidence and nature of adverse gynaecological events. The clinical records of all women admitted with gynaecological conditions over 9 months were assessed for adverse events, defined as an injury caused by medical management rather than the underlying disease. Adverse events occurred in 11.7% of admissions and 52% were avoidable. The majority of adverse events were minor (disability lasting less than 6 months). Mortality accounted for 17.7% of adverse events and 2.1% of all admissions. The death rate was higher in the elective admissions. The frequency of adverse events increased with age, the presence of co-existing illnesses, and severity of the illness on admission. Therapeutic mishaps (n = 67; 8%) were the most common type of adverse event. Lessons learnt from audit should reduce adverse events considerably. PMID- 16263541 TI - Ovarian cysts during pregnancy: dilemmas in diagnosis and management. AB - With routine obstetric ultrasound examinations, ovarian cysts are now more commonly diagnosed during pregnancy and their management is still a challenging clinical issue among obstetricians. The aim of this paper is to review the different management options of ovarian cysts detected during pregnancy, and it highlights the emergence of laparoscopic technique as a valuable therapeutic tool in these conditions. The review shows that conservative management and ultrasound follow-up is sufficient for the majority of cases of ovarian cysts during pregnancy. It also discusses the limitations of tumour markers during pregnancy, and the controversy around ultrasound-guided aspiration procedures. Emerging evidence suggests that if surgery is necessary, then it is preferable to perform an elective laparoscopic procedure at 16 - 23 weeks' gestation. It is time to consider establishing a UK registry to monitor the treatment offered to those women and to develop national guidelines to help in the management of this controversial issue. PMID- 16263543 TI - Laparoscopic laser sterilisation: an alternative option. AB - The aim of this prospective study was to evaluate the safety and efficacy of female laparoscopic laser tubal sterilisation. A total of 265 women underwent laparoscopic laser sterilisation as a day-case procedure at Princess Royal University Hospital in Kent between 1996 and 2001. The fallopian tube was divided at the isthmic portion using a neodymium-yttrium aluminium garnet (Nd:YAG) laser probe. All procedures were completed laparoscopically and patients were discharged within 6 h of surgery. No perioperative complications were encountered. The mean follow-up duration was 36 months (range 2 - 7 years) and no intra- or extrauterine pregnancies were reported throughout the entire follow-up period. We conclude that laparoscopic Nd:YAG laser sterilisation appears to be a safe and effective day-case method of female sterilisation. Larger studies with longer follow-up are needed to further define its role as a reliable long-term contraceptive method. PMID- 16263544 TI - Microbial isolates and HIV infection in couples attending fertility clinics in Sagamu, Nigeria. AB - This observational study was designed to determine the prevalence of HIV infection and the microbial isolates from the genital tracts of couples attending the fertility clinics of Olabisi Onabanjo University Teaching Hospital, Sagamu, Ogun State, Nigeria between January 2001 and December 2002. A total of 110 women and 49 of their male partners were recruited into the study. The majority of the patients were between 25 and 34 years (women: 71.8%; partners: 65.3%), though the men were significantly older than the women (p < 0.001). The overall prevalence of HIV infection in those who consented to screening was 8.2%, which was over twice the reported prevalence among the general population in Ogun State, Nigeria during the study period. Candida albicans (32.5%) and Staphylococcus aureus (27.5%) were the most frequently isolated microorganisms from the endocervix while Trichomonas vaginalis (37.9%) and Staphylococcus aureus (24.1%) were the most common microbes isolated from the posterior vaginal fornix. Of all the seminal qualities, only the volume showed a significant difference between the infected and non-infected samples (p < 0.004). This study suggests a higher prevalence of HIV infection among the infertile couples in our environment and it may be advisable to have them screened for HIV in the face of the present HIV situation in sub-Saharan Africa. PMID- 16263545 TI - In-vitro fertilisation treatment: factors affecting its results and outcome. AB - The objective of this study was to determine factors affecting results and outcome of in-vitro fertilisation (IVF). In this retrospective study, a total of 891 infertile women underwent IVF/ICSI cycles at the King Hussein Medical Center (KHMC) between January 2001 and December 2002. Conventional IVF treatment was performed in 64.6% of women and intracytoplasmic sperm injection (ICSI) in 35.4%, using a standardised long luteal protocol. Pregnancy rate was analysed according to age, type of infertility, cause of infertility, duration of infertility, number of eggs collected and follicle stimulating hormone (FSH) levels. A total of 126 cycles (14.1%) were cancelled. Among 765 cycles continued, fertilisation rate was 73.9%, implantation rate was 15.1% and pregnancy rate was 29.8%. Pregnant women had a multiple pregnancy rate of 28.9%, abortion rate of 13.6% and ectopic pregnancy rate of 1.3%. Duration and type of infertility had no significant effect on the pregnancy rate. Factors which appear to affect significantly the outcome of treatment include the woman's age, cause of infertility, basal concentrations of FSH, adequate ovarian responsiveness and the number of eggs collected. In some cases with poor outcome, the understanding of these factors may predict the results and lead to the development of new strategies to improve the outcome of IVF treatment. PMID- 16263546 TI - Primary fallopian tube carcinoma - the experience of a UK cancer centre and a review of the literature. AB - Primary fallopian tube carcinoma (PFTC) is rare but may be under-diagnosed. We have analysed the incidence, clinical findings and outcome in patients with PFTC at the RUH Gynaecological Cancer Centre in Bath between 1999 and 2004, and compared the incidence with that of advanced ovarian carcinoma (OC). Eight patients had PFTC, seven of whom were diagnosed after 2001, and 55 patients had advanced OC. Our data suggest a relative increase in the number of patients with PFTC over the study period. PFTC patients had a mean age of 69.6 years, most presented with postmenopausal bleeding, two had a second carcinoma, three were nulliparous and none were diagnosed pre-operatively. All were treated surgically and received platinum-based chemotherapy. Although PFTC patients had better outcomes than those with advanced OC, the difference was not statistically significant (p = 0.088). Accurate diagnosis and differentiation of PFTC from advanced OC are important for monitoring trends in incidence, for better characterisation of prognostic features and improved management. PMID- 16263547 TI - Is laparoscopic or abdominal hysterectomy with bilateral salpingo-oophorectomy more efficient in operative treatment of endometrial cancer? AB - We compared the efficiency and safety of 12 laparoscopic and 13 abdominal hysterectomies performed with bilateral salpingo-oophorectomy in the operative treatment of endometrial cancer. In all patients, the surgery was extended to include pelvic lymphadenectomy, where the histopathological examination confirmed advanced disease greater than IB (according to FIGO). Our work confirms that laparoscopic hysterectomy with bilateral salpingo-oophorectomy in endometrial cancer is a safe procedure for the patient and it seems to be more effective in lymph node sampling to abdominal surgery (via laparotomy). Moreover, complications are less frequent and severe when the patient is operated via the laparoscopic approach. PMID- 16263548 TI - Invasive cervical cancer and human immunodeficiency virus (HIV) infection in KwaZulu-Natal, South Africa. AB - The human immunodeficiency virus (HIV) can be expected to influence the course of disease and response to treatment of invasive carcinoma of the cervix. The extent and nature of this influence, however remains largely unknown. We therefore undertook a retrospective analysis of patients with carcinoma of the cervix at a tertiary referral centre in an African setting where HIV prevalence is high. There were 271 patients seen during a period of 1 year. Of these, 45 of the 206 tested were HIV infected (21.8%). The corresponding HIV prevalence for antenatal attendees was 38.7% in the region. The HIV-infected patients had lower mean haemoglobin levels and body mass indices than the HIV-non-infected women and were on average 13 years younger (p < 0.001), but otherwise did not differ with respect to demographics or disease parameters. They were, however, less likely to complete planned treatment. CD4 counts were below 200 in only 6 (21%) of 29 women tested. HIV-infected women in the African setting present with carcinoma of the cervix at a younger age, but the same disease stage as HIV-non-infected women, and without evidence of advanced immunocompromise. Circumstantial evidence is put forward by the study to suggest a more rapid decline in health and earlier demise for HIV-infected women with carcinoma of the cervix. PMID- 16263549 TI - The significance of plasma cells in the endometrium. PMID- 16263550 TI - Cushing's syndrome in pregnancy. PMID- 16263551 TI - Abdominal pain and hyperuricaemia in pregnancy. PMID- 16263552 TI - Percutaneous closure of an atrial septal defect during pregnancy using an Amplatzer occlusion device. PMID- 16263553 TI - Atraumatic fracture of the sacrum in pregnancy. PMID- 16263554 TI - Cervical placenta increta: a rare cause of near-fatal secondary postpartum haemorrhage. PMID- 16263557 TI - Management of interstitial (cornual) pregnancy at 17 weeks' gestation: conservation of a ruptured uterus. PMID- 16263555 TI - Treatment dilemmas after gonadectomy in complete androgen insensitivity syndrome. PMID- 16263556 TI - Heterotopic pregnancy: still a diagnostic dilemma. PMID- 16263558 TI - Endometrial carcinoma in a young obese patient. PMID- 16263560 TI - Lipoid cell tumour of the ovary: a rare cause of virilisation. PMID- 16263559 TI - Transitional cell carcinoma of the ovary. PMID- 16263561 TI - An unusual presentation of sebaceous gland hyperplasia of the vulva. PMID- 16263562 TI - Vulval sarcoid: a systemic presentation of sarcoidosis. PMID- 16263563 TI - Recurrent vulval leiomyoma in a postmenopausal patient mimicking vulval carcinoma. PMID- 16263564 TI - Uncommon metastatic site from vulval carcinoma: the tongue. PMID- 16263565 TI - Unusual presentation of recurrent extramammary Paget's disease. PMID- 16263567 TI - Foreign objects of long duration in the adult vagina. PMID- 16263566 TI - Successful treatment of a patient with gestational trophoblastic neoplasm metastatic to the colon. PMID- 16263569 TI - Mutant FLT3 signaling contributes to a block in myeloid differentiation. AB - FLT3 is a member of the class III receptor tyrosine kinase family and is primarily expressed on hematopoietic stem/progenitor cells. Somatic mutations of FLT3 involving internal tandem duplication (ITD) of the juxtamembrane domain or point mutations in the activation loop have been identified in approximately 17 - 34% and 7 - 9% of acute myeloid leukemia (AML) patients, respectively. The ITD mutations appear to activate the tyrosine kinase domain through receptor dimerization in a FLT3 ligand-independent manner. Constitutively activated FLT3 provides cells with proliferative and anti-apoptotic advantages and portends an especially poor prognosis for patients with this mutation. FLT3/ITD mutations also contribute to a block of myeloid differentiation. FLT3 tyrosine kinase inhibitors suppress the growth and induce apoptosis and differentiation of leukemia cells expressing FLT3/ITD mutants. Therefore, FLT3 is a therapeutic target and inhibition of FLT3 tyrosine kinase activity may provide a new approach in the treatment of leukemia carrying these mutations. PMID- 16263570 TI - ZAP-70 in B cell malignancies. AB - ZAP-70 has emerged as a protein of potential prognostic importance in chronic lymphocytic leukemia (CLL) following gene expression profiling which compared the 2 well established prognostic sub-sets, those with unmutated and mutated IgVH genes. This protein tyrosine kinase (PTK), known to be of importance in T and NK cell signaling but absent in normal peripheral B cells, is expressed in the majority of the poorer prognosis unmutated CLL and absent in most cases with mutated IgVH genes. ZAP-70 has been shown to be functionally important in the CLL cases in which it is expressed; it is also important in B cell development in mice and there is preliminary evidence for its expression in human B cell progenitors and activated B cells. Whether its expression in a sub-set of CLL cases is a result of a more activated cell type or a reflection of the stage of maturation of the transforming event(s) in CLL is open to debate. ZAP-70 is expressed in a minority of other B cell tumors but correlation with IgVH gene mutational status is lacking. The problems with ZAP-70 measurement, which has yet to be standardized, are reviewed together with its current status as a prognostic marker in CLL. PMID- 16263572 TI - Quality of life in chronic lymphocytic leukemia: a neglected issue. AB - Improvement in quality of life (QoL), together with overall survival and disease free survival, is a relevant endpoint for patients affected by chronic lymphocytic leukemia (CLL), a disease still considered as not curable. In addition, the study of the QoL can significantly contribute to investigate particular aspects related to different treatments which generally are not taken into account in clinical trials. A comprehensive approach to CLL should include also in the day-by-day practice the development of an appropriate and friendly interaction between the physician and patients aimed at improving the process of adaptation encompassing either the 'watch and wait' phase or the treatment period. The present review points out the role of QoL in the global patient management and care of CLL patients also in view of changes in the philosophy of treatment we have witnessed nowadays. PMID- 16263573 TI - Infradiaphragmatic versus supradiaphragmatic Hodgkin lymphoma: a retrospective review of 1,114 patients. AB - Infradiaphragmatic Hodgkin lymphoma (IDH) accounts for 4-13% of cases of stage I II Hodgkin lymphoma (HD). It has been associated with distinct pre-treatment characteristics and outcomes when compared with supradiaphragmatic HD (SDH). The comparison of IDH vs SDH can only be made in early and intermediate stages (I II), such a comparison is not possible for advanced stages (III-IV). This study retrospectively compared two groups of 1013 patients with stage I-II SDH and 101 patients with IDH (10%). These two sub-groups of patients were treated in 1988 1993 in 2 prospective randomized clinical trials in Germany for early and intermediate stages of Hodgkin lymphoma. IDH-patients were older (median 39 vs 31 years; p < 0.001), predominantly male (73% vs 52%; p < 0.001) and more often had involvement of 3 lymph node areas (LNA) (80% vs 55%; p < 0.001). Histology in IDH was more likely to be mixed cellularity (46.5% vs 23.6%, p < 0.001) or lymphocyte predominant (20 vs 10%, p = 0.003) and less likely nodular sclerosis (25% vs 63%, p < 0.001). In early-stage unfavorable disease, IDH was associated with a higher treatment failure rate (unadjusted hazard ratio 2, 95% CI, 1.3-3.4; p = 0.003). After controlling for age, sex, stage, histology, B-symptoms and involvement of 3 LNA, the adjusted hazard ratio was 1.25 (95% CI, 0.65-2.4; p = 0.51) so that IDH was no longer associated with a statistically significant treatment failure rate. Poorer outcomes with IDH as compared to SDH are attributable to its association with known adverse prognostic risk factors, but IDH, in itself, is not an independent adverse prognostic factor for treatment failure or survival. PMID- 16263571 TI - Significance of macrophage inflammatory protein-1 alpha (MIP-1alpha) in multiple myeloma. AB - Macrophage inflammatory protein-1 alpha (MIP-1alpha) is a member of the CC chemokine family and is primarily associated with cell adhesion and migration. It is produced by myeloma (MM) cells and directly stimulates osteoclast formation and differentiation in a dose dependent way. MIP-1alpha protein levels were elevated in the bone marrow plasma of MM patients and correlated with disease stage and activity. MIP-1alpha was also elevated in the serum of myeloma patients with severe bone disease and correlated positively with bone resorption markers providing evidence for a causal role of MIP-1alpha in the development of lytic bone lesions in MM. MIP-1alpha has also been found to stimulate proliferation, migration and survival of plasma cells. Mice, which were inoculated with myeloma cells and treated with a monoclonal rat anti-mouse MIP-1alpha antibody, showed a reduction of both paraprotein and lytic lesions. In addition, MIP-1alpha enhanced adhesive interactions between myeloma and marrow stromal cells, increasing the expression of RANKL and IL-6, which further increased bone destruction and tumor burden. Myeloma patients with high MIP-1alpha serum levels have poor prognosis. The positive correlation between MIP-1alpha and beta(2)-microglobulin that has been observed in MM patients at diagnosis further supports the notion that MIP 1alpha is not only a chemokine with osteoclast activity function but is also implicated in myeloma growth and survival. Therefore, MIP-1alpha pathway may serve as a target for the development of novel anti-myeloma therapies. PMID- 16263574 TI - Primary paranasal sinus lymphoma: natural history and improved outcome with central nervous system chemoprophylaxis. AB - Non-Hodgkin's lymphoma of the paranasal sinus is an uncommon presentation of extranodal lymphoma. Its natural history, treatment and prognosis have been infrequently characterized in the medical literature; however, a tendency to involve the central nervous system (CNS) has been noted. In British Columbia (population 4 million), a central database for lymphomas has allowed us to accurately track cases of paranasal sinus lymphoma diagnosed since 1980. A retrospective review was performed on the 44 patients who presented with primary paranasal sinus lymphoma (stage I or II) between 1980 and 1999. Histologic features were identified and immunophenotypic classification performed. Complete diagnostic and follow-up data including stage, treatment, response rates, sites of relapse and survival data were available for all patients. There were 26 men and 18 women. The types of lymphoma found were: diffuse large B cell (including immunoblastic), n = 37 (84%); T/NK nasal type, n = 3 (8%); peripheral T cell, not otherwise classified, n = 2 (4%); and others, n = 2 (4%). The median age at presentation was 66 years (range 27-97 years). The median follow-up for living patients was 114 months. For all 44 patients, the 5- and 10-year overall survivals were 48% and 41% and the disease-specific survivals 62% and 62%, respectively. Beginning in May 1985, intrathecal chemotherapy was added to our standard treatment plan of multi-agent chemotherapy and local irradiation. Before 1985, 2 of 5 patients developed leptomeningeal metastasis. Following the institution of intrathecal chemotherapy, only 8% (3 of 39) of patients have developed CNS disease. Introduction of intrathecal chemoprophylaxis was also associated with an improvement in overall survival from 20% to 51% and disease specific survival from 40% to 65%. Primary paranasal sinus lymphoma is an uncommon presentation of lymphoma that carries the potential risk of spreading to the leptomeninges. Treatment with combined modality chemotherapy and irradiation can cure many patients and the addition of intrathecal chemotherapy may reduce the risk of CNS relapse. PMID- 16263575 TI - Histiocytosis following T-acute lymphoblastic leukemia: a BFM study. AB - The coincidence of T-cell acute lymphoblastic leukemia (T-ALL) and histiocytic disorders, including hemophagocytic lymphohistiocytosis (T-ALL/HLH) and Langerhans cell histiocytosis (T-ALL/LCH), is very seldom and is usually associated with a dismal prognosis. Retrospective statistical analysis of all T ALL patients, who have been registered in the BFM-ALL trials from 1981 - 2001 and who have subsequently developed a LCH/HLH, in order to identify any common risk factors pre-disposing to the synchronous occurrence of both disorders. Six out of 971 T-ALL patients had either HLH or LCH ( approximately 0.03% of treated T ALL/year). The mean age at diagnosis of T-ALL/HLH/LCH was significantly lower than in the remaining T-ALL group (4.05 +/- 0.59 vs 8.82 +/- 0.14 years; p = 0.000). The mean initial leukocyte count was higher than in the non-HLH/LCH group (270,700 +/- 60,677 microl(-1) vs 134,141 +/- 5,663 microl(-1); p = 0.074). No hemophagocytosis was seen in the initial bone marrow (BM) smears. Five of 6 patients obtained a good prednisone response (GPR) at day 8 in peripheral blood with <5% blasts at day 15 in BM and all cases were in complete remission (CR) at day 33. The mean time until development of the histiocytosis was 17.95 months (range 2.5 - 33 months). Four patients developed a HLH and 2 a LCH. All patients with HLH showed a multi-organ involvement, while the LCH patients had only local disease. Only the LCH patients survived, while all patients with HLH died. The authors recommend a close follow-up for at least 3 years after diagnosis in younger T-ALL patients with high initial leukocyte count. PMID- 16263576 TI - IMVP-16/Pd followed by high-dose chemotherapy and autologous stem cell transplantation as a salvage therapy for refractory or relapsed peripheral T-cell lymphomas. AB - The present study aimed to analyse the treatment outcome of IMVP-16/Pd (ifosfamide, methotrexate, etoposide and prednisone) followed by high-dose chemotherapy and autologous stem cell transplantation (HDC/ASCT) for patients with peripheral T-cell lymphomas (PTCLs) who were previously treated with CHOP. Since 1995, 32 PTCL patients were treated with IMPV-16/Pd. Nine of 32 patients achieved a response (5 demonstrating complete response (CR) and 4 partial response), with an overall response rate of 28.1% (95% onfidence interval 0.12 0.45). Considering histopathologic subtypes, 3 of 4 relapsed natural killer (NK)/T-cell lymphoma patients (75%) achieved CR, but only 1 of 6 in non-NK/T-cell lymphoma patients (16.7%) achieved CR (P = 0.19). Six of 9 IMVP-16/Pd sensitive patients underwent HDC/ASCT. Three of them relapsed after 3, 4 and 15 months, respectively, of HDC/ASCT. Estimated 3-year overall survival and progression-free survival rates were 14.2% and 12.2%, respectively. Multivariate analysis revealed that responsiveness to first-line CHOP was a significant prognostic factor (P < 0.05). These results indicate that IMVP-16/Pd followed by HDC/ASCT appears to be an effective salvage regimen, especially for NK/T-cell lymphoma. PMID- 16263577 TI - The significance of carboxy-terminal telopeptide of type I collagen (ICTP) and osteocalcin (OC) in assessment of bone disease in patients with multiple myeloma. AB - Bone disease occurring in multiple myeloma is usually evaluated using radiological methods. These methods, however, provide not much information about the dynamic process of bone resorption and formation. This study analysed levels of serum markers of bone turnover (ICTP and OC), reflecting function of osteoclasts and osteoblasts. It demonstrates increased level of ICTP in 75 patients with MM compared to control group (8 persons) and patients with Waldenstrom's macroglobulinemia (10 persons). The level of ICTP was also higher in patients with more advanced bone disease and probably in higher stage of disease according to Salmon and Durie classification. This tendency was not observed in relation to OC. Result of the research confirms that ICTP may incur sensitive and specific markers of bone lesions in multiple myeloma. PMID- 16263578 TI - Immune recovery after conventional and non-myeloablative allogeneic stem cell transplantation. AB - The immune recovery of 66 patients undergoing allogeneic stem cell transplantation with either conventional or non-myeloablative conditioning regimen was studied. Infections post-transplant were enumerated and quantitative immunoglobuilins (IgG, IgA, IgM) and lymphocyte sub-sets 3, 6 and 12 months post transplant were measured. A significant difference was found in the immunologic recovery of non-myeloablative and conventional ASCT in the patient population. The T-helper cell reconstitution was significantly faster after NMA than conventional transplantation and the recovery of B cells was faster after conventional transplantation. Regarding immunoglobulin levels, a faster recovery of IgM levels after NMA-ASCT and a delayed recovery of IgA levels was observed in both groups. These were accompanied by a significant difference in the frequency and severity of infectious episodes. PMID- 16263579 TI - Elevated plasma levels of vascular endothelial growth factor is associated with marked splenomegaly in chronic myeloid leukemia. AB - Recent investigations support the idea that angiogenesis is involved in the pathophysiology of hematologic malignancies, including chronic myeloid leukemia (CML). The aim of the present study was to evaluate plasma levels of VEGF and bFGF in a cohort of 51 chronic-phase CML patients at the time of diagnosis, as well as to investigate the effect of imatinib therapy on VEGF amounts in CML patients. Plasma VEGF levels were significantly higher in patients studied as compared with the 20 healthy subjects (p<0.001), the median plasma VEGF level detected in patients analysed and healthy controls was 433.4 pg mL(-1) (range 65.2-2,452.7 pg mL(-1)) and 81.6 pg mL(-1) (range 44.2-338.7 pg mL(-1)), respectively. On the other hand, no difference in bFGF plasma levels could be found between chronic-phase CML patients and the control group. There were significant associations between plasma VEGF levels and some characteristics of patients evaluated, with trends for higher VEGF values in patients with enlarged spleens (p = 0.02) and those with higher platelet count (p<0.001). Of the patients, 11 received imatinib treatment. The initial VEGF levels markedly decreased after 6 months of imatinib therapy in each patient (p<0.001). These data support the important pathophysiological role of VEGF in CML. Further studies aiming to explore the detailed angiogenic profile of CML may help in developing new therapeutic strategies for this myeloproliferative disorder. PMID- 16263580 TI - Expression of Akt (protein kinase B) and its isoforms in malignant lymphomas. AB - Akt (protein kinase B) is a serine/threonine kinase involved in the regulation of cell survival signals. Akt is expressed in T- and B-lymphocytes and is activated in response to cytokine and antigen-receptor stimulation. Three isoforms of Akt have been identified, Akt-1, -2 and -3, but the expression pattern and specific functions of each have not yet been determined for many cell types. To determine whether Akt signaling is enhanced in human malignant lymphomas and to analyse the expression pattern of Akt isoforms in these neoplasms, Akt-1, -2 and -3 expression was studied in 38 cell lines derived from hematopoietic neoplasms, by RT-PCR and western blot analysis. The level of phosphorylated (active) Akt was also analysed in cell lines as well as in 72 human malignant non-Hodgkin's lymphoma tissues. The results suggest that there is constitutive activation of Akt in the majority of primary human lymphomas and hematopoietic cell lines and support its proposed key role in lymphoma cell survival. PMID- 16263581 TI - Concurrent administration of granulocyte colony-stimulating factor or granulocyte monocyte colony-stimulating factor enhances the biological activity of rituximab in a severe combined immunodeficiency mouse lymphoma model. AB - A predominant percentage of the in vivo antitumor activity of rituximab occurs through antibody-dependent cellular cytotoxicity (ADCC) via FcgammaRIII receptors. Co-expression of CD11b/CD18 (MAC-1), an adhesion molecule present in activated neutrophils, plays an important role in the induction of ADCC. The effects of granulocyte-monocyte colony-stimulating factor (GM-CSF) and granulocyte colony-stimulating factor (G-CSF) on the biological activity of rituximab were studied in a non-Hodgkin's lymphoma (NHL)-bearing severe combined immunodeficiency (SCID) mouse model. Natural killer (NK) cell-depleted SCID mice were inoculated intravenously with Raji cells. Animals were divided into 6 cohorts: group A: placebo (saline injection); group B: murine (m)-G-CSF; group C: m-GM-CSF; group D: rituximab alone; group E: concurrent m-G-CSF and rituximab; and group F: concurrent m-GM-CSF and rituximab. Treatment with G-CSF or GM-CSF led to a 1.5- to 2-fold increase of CD11b/CD18 expression in neutrophils. Treatment with G-CSF led to the highest expression of CD11b/CD18 on neutrophils. No antitumor activity was observed among mice treated with G-CSF or GM-CSF alone. After 3 months, survival rates were highest in animals treated with rituximab and G-CSF (53.3%) compared to rituximab alone (13.3%) or in combination with peg-GM CSF (26.7%). Increasing neutrophil counts via cytokine stimulation may play an important role in augmenting rituximab-associated antitumor activity. PMID- 16263582 TI - Activation of CD44 facilitates DNA repair in T-cell lymphoma but has differential effects on apoptosis induced by chemotherapeutic agents and ionizing radiation. AB - Expression of CD44s (standard form) in malignant lymphoma is a poor indicator of survival. To investigate whether activation of CD44s can protect from cell death, this study compared the extent of apoptosis induced by chemotherapeutic agents and ionizing radiation (IR) on T-lymphoma cell lines in the presence or absence of adherent hyaluronan and monoclonal antibodies (MoAbs). Growth in the presence of adherent ligands enhanced apoptosis induced by dexamethasone (Dex), but protected cells from epirubicin-induced apoptosis. In IR-induced apoptosis, mouse lymphoma cells had resistance against apoptosis when treated with hyaluronan (HA), although acute cell death reached the same plateau regardless of treatment with adherent MoAbs in human lymphoma cell line. However, the post-irradiated repopulation of lymphoma cells was strikingly accelerated in those treated with CD44 adherent ligands. This repopulation process correlated with the remarkable upregulation of proliferating cell nuclear antigen (PCNA), which is a protein involved in DNA repair. Unscheduled DNA synthesis (UDS), a measure of DNA repair, was consistently enhanced in CD44s-stimulated cells after exposure to radiation. The results suggest that the poor prognostic indication of CD44 expression is more a consequence of enhanced DNA repair following genotoxic damage than of direct resistance to apoptosis. PMID- 16263583 TI - A marine fish diet reduces spontaneous lymphoma in outbred Swiss-Webster mice. AB - Diets rich in marine organisms or their oils are known to suppress solid tumor development in humans and rodents, but the potential for marine foods to affect hematopoietic system cancers is not well understood. As part of a toxicology study, we fed groups of mice three different diets for 10 weeks: marine fish, 58% homogenized Atlantic smelt and herring; freshwater fish, 58% smelt and alewife from the North American Great Lakes, and commercial dry rodent chow. Between 1 and 15 weeks following dietary treatment, 20 of 103 (19.4%) mice unexpectedly developed spontaneous lymphoma. Disease incidence peaked when the mice were 7-8 months old, and was not distributed equally across treatment groups. Mice in the control (30%) and fresh water fish (27.5%) groups had significantly higher incidences of lymphoma than those fed Atlantic fish species (5%). Although our experiment was not originally designed for this purpose, our results indicate that consumption of fat-rich Atlantic smelt and herring protected mice against hematopoietic tumor development. PMID- 16263584 TI - Variant translocation with a deletion of derivative (9q) in a case of Philadelphia chromosome positive (Ph +) essential thrombocythemia (ET), a variant of chronic myelogenous leukemia (CML) with a poor prognosis. AB - Patients presenting with thrombocytosis require thorough clinical and laboratory evaluation to determine whether they suffer from essential thrombocythemia or another myeloproliferative disorder. This distinction becomes increasingly relevant as targeted agents become available to treat specific myeloproliferative diseases. Cytogenetic testing plays a major role in this analysis. This study presents a patient with Philadelphia chromosome positive (Ph + ) thrombocytosis and a cryptic der(9q)t(5;9)t(9;22) not found by conventional cytogenetics, whose disease progressed within 2 years to typical myeloblastic crisis of CML. It discusses the entity of Ph + ET, the utility of molecular cytogenetic testing in the diagnosis of this unusual disease entity and the importance of cytogenetic testing in the prognosis of ET. PMID- 16263586 TI - Simultaneous presentation of acute monoblastic leukemia and mantle cell lymphoma: case report and review of the literature. AB - This paper reports a 73-year old woman with simultaneous presentation of acute monoblastic leukemia (acute myeloid leukemia (AML), French-American-British (FAB) type M5a) and mantle cell lymphoma. The patient presented with wasting, generalized lymphadenopathy, an extensive infiltrative rash and pancytopenia. Bone marrow and lymph node histopatholology showed extensive infiltration by leukemic monoblasts. Marrow cytogenetics revealed a complex karyotype, including t(8;16)(p11;p13). Flow cytometric immunophenotyping of peripheral blood, lymph node and bone marrow demonstrated two populations, expressing CD5, CD19, CD20 and CD22 and CD45, HLA-DR, CD13, CD33, CD14 and CD38, respectively. A focus of abnormal lymphocytes in the lymph node biopsy demonstrated BCL1 expression and t(11;14)(p11;p13) by fluorescence in situ hybridization and immunoglobulin heavy chain gene rearrangement by the polymerase chain reaction. The patient received infusional cytarabine, daunorubicin and etoposide chemotherapy, with complete remission of both the AML and the mantle cell leukemia. To the authors' knowledge, this is the first report of simultaneous presentations of AML, FAB M5a and mantle cell lymphoma. The case is discussed and the literature is reviewed. PMID- 16263585 TI - Toxic epidermal necrolysis associated with denileukin diftitox (DAB389IL-2) administration in a patient with follicular large cell lymphoma. AB - Denileukin diftitox (DAB(389)IL-2 or Ontak) is a synthetic fusion protein with demonstrated efficacy in a number of lymphoproliferative disorders, including non Hodgkin's lymphoma. We report the case of a 45-year-old man with progressive follicular large cell lymphoma following an autologous stem cell transplant treated with denileukin diftitox who developed a fatal skin rash associated with extensive erythema, edema and large bullae involving his entire body. The clinical features and pathology were compatible with toxic epidermal necrolysis. This is the first reported case of toxic epidermal necrolysis in the literature associated with denileukin diftitox. PMID- 16263587 TI - Heart and muscle involvement by extra-medullary myeloid leukemia: a case report and review of the literature. AB - Extra-medullary myeloid tumours (EMT) have been described after curative treatment for acute myeloid leukaemia (AML) in increasing numbers after allogeneic stem cell transplantation. The sites of manifestations are ubiquitous and the discovery is most frequently guided by symptoms reported by the patient or by findings on clinical examination. This study reports a case of EMT in muscles and the heart 1.5 years after allogeneic transplantation for an AML with t(8;21)(q22;23) who achieved a complete remission by use of an idarubicine-based combination chemotherapy. Pathological and imaging findings are presented and treatment options are discussed. PMID- 16263588 TI - Hypercalcemia due to all trans retinoic acid in the treatment of acute promyelocytic leukemia potentiated by voriconazole. AB - All-trans-retinoic acid (ATRA) is a new and effective treatment of acute promyelocytic leukemia. It has many side-effects, including the retinoic acid syndrome and Sweet's syndrome. There have been only nine cases of hypercalcemia associated with ATRA described in the literature. We discuss a case of hypercalcemia, which we believe was due to inhibition of cytochrome P450 function by voriconizole when used concomitantly with ATRA. PMID- 16263589 TI - TLS/FUS-ERG fusion gene in acute lymphoblastic leukemia with t(16;21)(p11;q22) and monitoring of minimal residual disease. AB - This study reports a 1-year-old boy with precursor B cell acute lymphoblastic leukemia (ALL) carrying t(16;21)(p11;q22). Reverse transcriptase-polymerase chain reaction (RT-PCR) and direct sequence analysis showed TLS/FUS-ERG chimeric mRNA with a novel junctional pattern of exon 7 of TLS/FUS and exon 6 of ERG. He did not respond to ALL-oriented therapy. Complete remission (CR) was achieved by chemotherapy oriented for acute myeloid leukemia. Allogenic bone marrow transplantation was done and he has been in CR for 24 months. TLS/FUS-ERG chimeric mRNA was not detected after CR. This is the first report of an ALL patient with a TLS/FUS-ERG fusion transcript. PMID- 16263590 TI - Low-dose thalidomide-induced agranulocytosis in a multiple myeloma patient treated at diagnosis. AB - Thalidomide represents a recent and innovative therapeutic approach in multiple myeloma. Main toxicity usually consists in somnolence, constipation, peripheral neuropathy and deep vein thrombosis, but, unlike alkylating agents, thalidomide is reported to rarely induce severe hematologic toxicity. The majority of patients developing neutropenia are heavily pretreated with three or more lines of chemotherapy. Here, we report, for the first time, clinical and laboratory data of a 66-year-old female patient with multiple myeloma at diagnosis who, after 4 weeks of thalidomide treatment, developed a grade 4 WHO neutropenia with septicemia. A brief review of the literature and suggestions for possible predictive factors of this toxicity are made. PMID- 16263591 TI - Efficacy and safety of oral fludarabine/cyclophosphamide regimen in previously treated indolent lymphomas. PMID- 16263592 TI - Prolonged remission of adult T-cell leukemia/lymphoma treated with interferon gamma following autologous peripheral blood stem cell transplantation. PMID- 16263593 TI - The pregnant spontaneously hypertensive rat as a model of asymmetric intrauterine growth retardation and neurodevelopmental delay. AB - INTRODUCTION: Hypertension in pregnancy and vascular placental insufficiency are considered common pathogenic factors in human intrauterine growth retardation (IUGR). IUGR neonates experience higher mortality, and the surviving infants have a higher incidence of neurological and intellectual impairment. METHODS: To mimic this condition, we used pregnant spontaneously hypertensive rats (SHR) and performed biometric measurements on Embryonic Day 20, postnatal developmental reflexes, and locomotor activity evaluations. RESULTS: SHR fetuses had significant decreased body weight compared to the Wistar-Kyoto control fetuses (1.51+/-0.02 g vs. 2.05+/-0.01 g, respectively; p<0.0001), and were relatively microcephalic (2.86+/-0.04 cm vs. 3.3+/-0.03 cm, respectively; p<0.0001). Their cephalization index (head circumference/body weight) was increased (1.88+/-0.03 vs. 1.62+/-0.02, respectively; p<0.0001), indicating a "brain-sparing" process. The disproportional ratio indicated that the IUGR type in this model is asymmetric. The SHR pups exhibited a significant (p<0.04) neurodevelopmental delay in the acquisition of neonatal reflexes (righting, negative geotaxis, placing), but they spontaneously caught up with the control pups after approximately 10 days. On Day 30, the SHR pups exhibited significantly increased walking speed and distance and spent less time in quadrant than the controls (p<0.002). CONCLUSION: We speculate that the model of pregnant SHR closely simulate human IUGR caused by hypertension in pregnancy and should enable investigation of mechanisms of hypertension-mediated placenta-vascular injury as well as provide a system for preclinical evaluations of future preventive neuroprotective treatments. PMID- 16263594 TI - Variation in the urinary protein/creatinine ratio at four different periods of the day in hypertensive pregnant women. AB - OBJECTIVE: To assess the urine protein/creatinine ratio in urine samples of pregnant women with hypertension in regard to: 1) the presence of significant variation at different periods of the day; 2) the differences if they exist, to identify the most reliable period of the day for sampling; and 3) whether the first sample, obtained when the patient arrives at the clinic, correlates with the same accuracy, with the 24-hour proteinuria. DESIGN: Cross-sectional study. PLACE: Obstetrics Emergency Department, Hospital de Clinicas de Porto Alegre, a teaching hospital in Porto Alegre, Brazil. POPULATION: Seventy-five women with hypertension with 20-week gestation or over. METHODS: Urine samples for determination of the protein/creatinine ratio were obtained on arrival (first specimen) and every 6 hours thereafter, totaling four samples in 24 hours. Four sampling periods were established: 1) from 8 am to 2 pm, 2) from 2 pm to 8 pm, 3) from 8 pm to 2 am, and 4) from 2 am to 8 am. The protein/creatinine ratio in the four different day periods were compared with the 24-hour proteinuria obtained simultaneously. The results were analyzed by the Spearman correlation and the receiver-operator characteristic (ROC) curve. RESULTS: The urine protein/creatinine ratio is strongly correlated (Spearman correlation equal to 0.8 or greater) with the 24-hour proteinuria at all four periods of the day (p<0.001), as well as the first sample obtained on arrival (p=0.003). These findings were corroborated by the ROC curve in which the values of four day periods and that of the first sample were equal to or greater than 0.930. CONCLUSION: In hypertensive pregnant women, the single voided urine sample protein/creatinine ratio, irrespective of sampling time, is strongly correlated with the 24-hour proteinuria, as is the sample obtained on arrival. PMID- 16263596 TI - Placental tissue levels of nonesterified polyunsaturated fatty acids in normal and preeclamptic pregnancies. AB - OBJECTIVE: We previously reported that maternal plasma levels of nonesterified polyunsaturated fatty acids were decreased in women with preeclampsia as compared to women with normal pregnancies. Polyunsaturated fatty acids of the n-6 and n-3 families are essential dietary fatty acids. The n-6 polyunsaturated fatty acids are involved in inflammatory reactions, whereas n-3 polyunsaturated fatty acids protect against inflammation. METHODS: In this study, we investigated the composition of nonesterified polyunsaturated fatty acids in placental tissue of normal and preeclamptic pregnancies. Linoleic (18:2) and arachidonic (20:4, AA) acids of the n-6 family and linolenic (18:3), eicosapentaenoic (20:5, EPA) and docosahexaenoic (22:6, DHA) acids of the n-3 family were analyzed. CONCLUSIONS: We found that total concentrations of nonesterified polyunsaturated fatty acids were lower in placental tissues from preeclamptic pregnancies than from normal pregnancies. Both n-6 and n-3 polyunsaturated fatty acids were decreased. The decrease in n-6 polyunsaturated fatty acids was due to a decrease in AA. The concentration of linoleic acid was not altered. For n-3 polyunsaturated fatty acids, DHA was decreased, with no change in linolenic acid. The percentage decrease in EPA and DHA was greater than for AA, so the ratio of n-6 to n-3 polyunsaturated fatty acids was two-fold higher in preeclampsia than normal pregnancy. Because the levels of essential fatty acids, linoleic (18:2) and linolenic (18:3), are preserved but levels of metabolites (AA and DHA) are reduced, we speculate that placental oxidative stress and increased conversion of AA to thromboxane are responsible for the decreased concentrations of polyunsaturated fatty acids in preeclampsia. PMID- 16263595 TI - Effect of preeclampsia on blood pressure in newborn very low birth weight infants. AB - OBJECTIVE: To test the hypothesis that very low birth infants born to mothers with preeclampsia have higher blood pressure over the first week of life than infants whose mothers did not have preeclampsia. METHOD: Infants born at<1,350 g who survived at least one week were stratified by gestational age ( or= 29 completed weeks) and grouped by the presence or absence of preeclampsia. Highest and lowest systolic and mean and diastolic blood pressures were recorded for each of the first seven days of life. Serial blood pressures were analyzed by repeated measures ANOVA: The presence of hypertension (defined as >or= 3 days with the highest systolic blood pressure>90th percentile for gestational age stratum and day-specific range) was analyzed by binary logistic regression. RESULTS: Infants >or= 29 weeks gestational age born to mothers with preeclampsia had higher blood pressures than did controls. Infants or= 29 weeks gestation. The long-term significance of this finding is not known. PMID- 16263597 TI - The correlation between renal function and systolic or diastolic blood pressure in severe preeclamptic women. AB - OBJECTIVE: To evaluate the correlation between renal function and systolic or diastolic blood pressure in preeclamptic mothers. METHODS: In this prospective study from August 1998 to September 2002, 28 women >or= 28 weeks gestation with severe preeclampsia were selected. Meanwhile, 56 normotensive pregnant women without proteinuria or edema served as the control group. Urine was collected for 24 hours for all subjects. The concentration of uric acid, blood urea nitrogen, creatinine, sodium, calcium, and albumin in the 24-hour urine and blood of both groups were examined. Neonatal outcome also was evaluated. RESULTS: The serum and 24-hour urine concentration of blood urea nitrogen, creatinine, and albumin were significantly higher in severe preeclamptic women. Serum uric acid and urinary albumin/creatinine ratio was significantly higher in severe preeclamptic women compared with that in normotensive mothers and showed positive correlation with systolic or diastolic blood pressure. On the other hand, serum calcium/creatinine ratio was significantly lower in the severe preeclamptic group and negatively correlated to blood pressure. In multiple regressions, systolic or diastolic blood pressure was dependent on serum uric acid, albumin/creatinine, and calcium/creatinine ratios. Fetal birth weight was significantly lower in women with severe preeclampsia and with a lower Apgar score < 7 at 1 minute and 5 minutes and more preterm delivery compared with that in normotensive women. CONCLUSION: Renal function in women with severe preeclampsia was significantly impaired and highly correlated with systolic or diastolic blood pressure. PMID- 16263598 TI - Severe preeclampsia is associated with a positive family history of hypertension and hypercholesterolemia. AB - OBJECTIVE: To investigate an association between a family history of cardiovascular disease and severe preeclampsia and/or HELLP syndrome (Haemolysis, Elevated Liver enzymes, Low Platelets). METHODS: One hundred twenty-eight women with a history of severe preeclampsia and/or HELLP syndrome and 123 women with previous uncomplicated pregnancies only were included in the study. All participants completed questionnaires about diagnoses of cardiovascular diseases, hypertension, and hypercholesterolemia among their first-degree relatives, which were subsequently confirmed by the relatives' general practitioners. The main outcome measures were the prevalence of cardiovascular diseases, hypertension, and hypercholesterolemia among first-degree relatives of both groups. Statistical analysis was done using chi(2)-analysis. RESULTS: The prevalence of familial cardiovascular disease among women with a history of severe preeclampsia and/or HELLP syndrome (23%) compared to controls (19%) was not significantly different (OR 1.3, 95%CI 0.7-2.5). However, women with a history of severe preeclampsia and/or HELLP syndrome more often had one or more first-degree relatives with hypertension and/or hypercholesterolemia before the age of 60 years compared to controls (54% vs. 32%, respectively; OR 2.6, 95%CI 1.5-4.3). The prevalence of hypertension and hypercholesterolemia among first-degree relatives, irrespective of age, also was significantly higher among women with a history of severe preeclampsia and/or HELLP syndrome as compared to controls (60% vs. 42%, respectively; OR 2.0, 95%CI 1.2-3.4). CONCLUSION: Severe preeclampsia is associated with a positive family history of hypertension and/or hypercholesterolemia. PMID- 16263599 TI - Mid-trimester placentation assessment in high-risk pregnancies using maternal serum screening and uterine artery Doppler. AB - OBJECTIVE: To determine the value of uterine artery velocimetry and mid-trimester maternal serum AFP/hCG measurements in predicting pregnancy complications in a high-risk group of pregnant patients. METHODS: Eighty-eight patients with chronic hypertension, previous preeclampsia, and thrombophilia were included. Maternal serum AFP/hCG was examined between 15-16 weeks gestation. Levels > 3 multiple of median (MoM) for hCG and > 2 MoM for AFP were considered abnormal. Color Doppler ultrasound was performed at 23-24 weeks gestation. Diastolic notching and pulsatility index (PI) above the 95th percentile were considered abnormal. RESULTS: Thirty-three patients had abnormal uterine artery waveform: 8 patients had abnormal maternal serum hCG and 5 had abnormal maternal serum AFP. Bilateral abnormal uterine artery waveform was associated with pregnancies complicated by lower gestational age at delivery (p=0.05) and birth weight (p<0.01), higher rates of preeclampsia (p=0.006), SGA (p=0.0001), preterm delivery (p=0.0001), and cesarean section rate (p<0.0001) in comparison to patients with normal uterine artery Doppler. Pregnant women with elevated hCG had higher rates of preeclampsia (p=0.006); preterm delivery (p=0.005) and SGA (P=0.03) and, lower birth weight (p=0.001). No significant differences were noted for clinical outcomes according to AFP data. Conclusions. Abnormal uterine artery waveform is superior to maternal serum hCG for identification of placental pathology leading to preterm delivery, low birth weight, and preeclampsia in high-risk pregnant patients. PMID- 16263600 TI - Patterns of pregnancy-related hypertension in black and white women. AB - OBJECTIVE: To determine whether the higher rate of preeclampsia previously reported in Black women is attributable to a higher rate of chronic hypertension in that group. METHODS: Rates and severity of pregnancy-related hypertensive disease in Black and White women with and without chronic hypertension were reviewed from the medical records of our institution. RESULTS: Of 1,355 records reviewed, evidence of hypertensive disease was noted in 101 singleton pregnancies. Hypertension during pregnancy occurred in 7.4% of Black and 7.4% of White women. Among women with hypertension in pregnancy but no chronic hypertension, Blacks were more likely to be diagnosed with preeclampsia (78 vs. 53%, p=0.04) and more likely to have had systolic blood pressures >160 mmHg (43 vs. 17%, p=0.01). However, the rates of severe preeclampsia were similar in Black and White women since Whites were more likely to be diagnosed with severe preeclampsia in the absence of very high systolic blood pressures. Black multiparous patients with hypertension were three times more likely than White multiparous patients to be diagnosed with preeclampsia (80 vs. 27%, p=0.01). Conclusions. In our population, Black women with hypertension in pregnancy in the absence of chronic hypertension were more likely to be diagnosed with preeclampsia than were White women. This finding suggests that the higher rate of preeclampsia among Black women is not completely explained by higher rates of chronic hypertension in that group. PMID- 16263602 TI - First trimester sex hormone-binding globulin and subsequent development of preeclampsia or other adverse pregnancy outcomes. AB - OBJECTIVE: To investigate whether first trimester maternal serum sex hormone binding globulin (SHBG) concentrations are altered in women who subsequently develop preeclampsia or other pregnancy complications. POPULATION: Women undergoing first trimester combined ultrasound and biochemical screening for chromosomal anomalies. We searched the database and identified 32 pregnancies resulting in miscarriage, 64 pregnancies with preexisting or gestational diabetes mellitus, 107 with fetal growth restriction, 103 with preeclampsia, 64 with pregnancy-induced hypertension, and 26 with spontaneous preterm delivery. We also selected 400 controls from among the population of pregnancies that had a delivery of a normal baby with no pregnancy complications. METHODS: Maternal serum SHBG concentrations were measured retrospectively using a competitive chemiluminescent immunoassay. The levels between those with normal outcome and those resulting in adverse outcome were compared. RESULTS: The median maternal serum SHBG concentration was not significantly different from controls, in those that subsequently developed preeclampsia (median MoM 1.05), non-proteinuric hypertension (median MoM 0.94) or preterm delivery (median MoM 1.15). The levels were significantly lower in those with diabetes (median MoM, 0.81 p=0.0005) and those pregnancies resulting in miscarriage (median MoM 0.80, p=0.008). CONCLUSION: First trimester maternal serum SHBG concentrations are no different from controls in women who subsequently develop preeclampsia, pregnancy-induced hypertension, fetal growth restriction, or preterm delivery. Levels are reduced in those who subsequently miscarry or in those presenting with diabetes. PMID- 16263601 TI - Urine protein estimation in hypertensive pregnancy: which thresholds and laboratory assay best predict clinical outcome? AB - OBJECTIVE: To determine what threshold for proteinuria could best predict clinical outcome and whether this threshold could be applied universally to any biochemical assay. DESIGN: A prospective observational study of hypertensive pregnancies referred for further assessment after in a UK University hospital (n=197). Twenty-four hour urine protein was measured by two different assays [benzethonium chloride assay (BCA) and Bradford assay]. The differences between the two assays were calculated from Receiver Operating Characteristic (ROC) curves. Commonly used thresholds for defining preeclampsia (0.3 and 0.5 g/24 hours) were explored for both assays for the prediction of adverse clinical outcomes (severe hypertension, Birthweight<10th percentile, preterm delivery, and a composite biochemical/haematological derangement). RESULTS: The two assays are not equivalent. The prevalence of>300 mg/24 hour proteinuria and, hence, the prevalence of preeclampsia differed between the two assays. ROC curve analysis demonstrates that the two assays are similar in terms of overall performance as predictive tests. However the threshold of 300 mg/24 hours performs poorly as a predictor of clinical risk. Likelihood ratios (LR) for the BCA at the 300 mg/L threshold for each clinical outcome do not achieve statistical significance. At the 500 mg/L threshold, the LR+for the BCA assay does achieve statistical significance for severe hypertension (LR+:1.51 95% CI 0.99-2.28) and for birthweight<10th percentile (LR+:1.72 95% CI 1.11-2.66). For the Bradford assay at the 300 mg/24 hour threshold, the LR+does achieve statistical significance for birthweight<10th percentile (LR+:1.71 95% CI 1.41-4.31). However, at the 500 mg/24 hour threshold, the LR+is significant for severe hypertension (LR+:2.15 95% CI 1.07-4.34), birthweight<10th percentile (LR+:2.79 95% CI 1.4-5.54) and biochemical disease (LR+:2.47 95% CI 1.22-5.01). CONCLUSIONS: This study suggests that thresholds for proteinuria need to be higher (possibly>or=0.5 g/24 hours) and there is the need for a "gold standard" proteinuria assay against which all other measures of quantification can be assessed. PMID- 16263604 TI - Seppo Santavirta, 1945-2005. PMID- 16263603 TI - Temporary resolution of preeclamptic symptoms after intrauterine death of one twin. AB - In a 34 year old woman with dichorionic twin pregnancy preeclampsia resolved after the intrauterine death of one of the HLA-identical twins and recurred with the growth of the placenta of the surviving twin later in pregnancy. This case gives indirect evidence that the clinical course of preeclampsia is a dose dependent phenomenon in conjunction with vital placental tissue. PMID- 16263605 TI - Biotribology. PMID- 16263606 TI - Current management: Management of rheumatic diseases in the era of biological anti-rheumatic drugs. PMID- 16263607 TI - Vertebroplasty and kyphoplasty: New treatment strategies for fractures in the osteoporotic spine. AB - During the last decade, two new treatment modalities for osteoporotic vertebral fractures have gained more interest: percutaneous vertebroplasty and kyphoplasty. The techniques and the short-term clinical results and complications have been presented but there is no scientific evidence-based information regarding the efficacy of the procedures, such as randomized controlled trials (RCT). Instead, we have to rely on prospective and retrospective uncontrolled short-term observational studies and case-control studies. These studies have shown consistently that the short-term results after the procedures are favorable as regards both pain relief and functional status. It is currently unknown, however, whether a vertebroplasty or a kyphoplasty gives a better outcome than nonoperative treatment, and whether the long-term results are as favorable as the short-term results. PMID- 16263609 TI - Immune response and effect of adenovirus-mediated human BMP-2 gene transfer on the repair of segmental tibial bone defects in goats. AB - BACKGROUND: Tissue-engineered bone may be used for filling bone defects. There are, however, no reports on this technique used in large animals. METHODS: We evaluated the effectiveness of, and immune response in repairing diaphyseal bone defects by gene transfer using bone morphogenetic proteins (BMPs). We used adenovirus-mediated human BMP-2 (Adv-hBMP-2) gene-transduced bone marrow stromal cells (BMSCs) to repair 2.1-cm segmental tibial bone defects in goats (group I, n = 7). An Adv-ssgal-transduced BMSC group (group II, n = 5), a non-transduced BMSC group (group III, n = 5), and an untreated group (group IV, n = 2) were used as controls. Self-secreted extracellular matrix was used as cellular carrier. RESULTS: Radiographic and histomorphometric examination demonstrated more callus in the bone defects of group I compared to other groups. Week 24 after implantation, the defect healing rates of groups I, II, III, and IV were 6/7, 1/5, 2/5, and 0/2, respectively. The maximum compressive strength of new tissue in the bone defects of group I was higher than those of groups II and III. Temporary cellular and persistent humoral immune responses against adenovirus were detected after hBMP-2 gene transfer. INTERPRETATION: We found that Adv-hBMP 2 genetransduced BMSCs had superior osteoinductivity in the repair of tibial bone defects in goats, but it could cause temporary cellular and persistent humoral immune responses against adenovirus. PMID- 16263608 TI - Implantation of perichondrium-derived chondrocytes in physeal defects of rabbit tibiae. AB - BACKGROUND: Various cells and scaffolds have been used experimentally as interposition materials after physeal bar resection to treat partial physeal growth arrest. However, results of these cell therapies are unpredictable. METHODS: We investigated the chondrogenic differentiation of rabbit perichondrial cells in vitro. We also investigated radiographic and histological changes after implantation of the perichondrium-derived chondrocytes, embedded in fibrin beads, into defects created in the medial aspect of the proximal tibial physis of 26 8 week-old New Zealand White rabbits. Physeal defects of the contralateral tibiae were left intact without any treatment, or were treated with fibrin beads not containing cells. RESULTS: The perichondrial cells cultured in the alginate- fibrin beads were positive in immunohistochemical staining for S-100 protein and the extracellular area was stained with Safranin-O. RNase protection assay showed that the expression of type II collagen and aggrecan continued In the cell and bead-treated tibiae, varus deformity was reduced significantly and tibial length was maintained longer than that of the contralateral tibia. Histologically, cartilage clusters of varying size with intervening micro-bony trabeculae were found in the cell and bead-treated tibiae. The lateral, remaining physis in the untreated tibiae showed altered histology. INTERPRETATION: Implantation of fibrin beads containing perichondrium-derived chondrocytes into the physeal defect helps to prevent angular deformation and shortening of long bone, but not completely. Physeal bony bar formation adversely affects the growth of the remaining physis from early stages onward. PMID- 16263611 TI - Serum total acid phosphatase for monitoring the clinical course of giant cell tumors of bone--26 patients with 5 local recurrences. AB - BACKGROUND: Giant cell tumor of bone (GCT) is a bone-destroying tumor that sometimes recurs locally after treatment. A recent study showed increased levels of serum total acid phosphatase (TACP). METHODS: We assessed TACP in the serum of 26 patients with primary GCT, and in 5 of them who developed a local recurrence. RESULTS: We found a correlation between TACP level in serum and tumor size. TACP levels that were elevated preoperatively in patients with GCT became normalized after surgery, but increased in 3 of the 5 patients with local recurrence. INTERPRETATION: TACP could be used as a tumor marker for monitoring response to treatment of GCT. PMID- 16263610 TI - Gas chromatography-mass spectroscopy analysis of emissions from cement when using ultrasonically driven tools. AB - BACKGROUND: Ultrasonically driven tools have been used to reduce the incidence of complications during cement removal at revision hip replacement operations. These have been shown to be safe and effective in various ways, but produce fumes. METHODS: Using gas chromatography-mass spectroscopy, we analyzed the fumes produced during the use of these ultrasonic tools for the removal of bone cement, both in the laboratory and during actual surgery. RESULTS: Benzene, styrene, methylmethacrylate, xylene, toluene, isopropyl alcohol and dichlorobenzene were some of the substances isolated from the fumes in the laboratory. Styrene and methylmethacrylate were the main components. Concentrations of all the above components taken from the breathing zone of the operating staff during actual surgery were well below the safety limits. INTERPRETATION: The use of ultrasonic tools for cement removal appears to be safe. PMID- 16263612 TI - Limited influence of prosthetic position on aseptic loosening of elbow replacements: 125 elbows followed for an average period of 5.6 years. AB - BACKGROUND: Aseptic loosening of elbow replacements, seen in long-term follow-up, remains a problem. In this study, we attempted to determine the influence of cementing technique, prosthetic position, different component sizes, use of a bone plug, and intraoperative fractures on the development and progression of radiolucent lines and aseptic loosening. METHODS: We studied standard radiographs of 125 primary Souter-Strathclyde total elbow prostheses using the Wrightington method. Additionally, 104 preoperative radiographs were available for analysis. We used a Markow statistical model to detect relationships between all factors described above. RESULTS: After a mean follow-up time of 5.5 (2-19) years, 21 (17%) prostheses had loosened radiographically (10-year survival: 65%). When the humeral component was tilted more medially or more anteriorly, we found development of radiolucent lines at the medial condyle and at the posterior side of the humeral component. However, the progression of these lines was not influenced by these positions. No other prognostic factors for radiolucent lines or aseptic loosening were found. INTERPRETATION: Despite the small number of elbows studied, the weak influence of prosthetic position on aseptic loosening gives more ground for a multifactorial cause for aseptic loosening of the Souter Strathclyde total elbow prosthesis. PMID- 16263613 TI - Direct medical costs of traumatic thoracolumbar spine fractures. AB - BACKGROUND: The costs and cost-effectiveness of treatment of thoracolumbar fractures are poorly known. METHODS: We estimated the costs of hospital care and outpatient visits for patients with traumatic thoracolumbar spine fractures. RESULTS: Stable fractures without neurological deficits were treated nonoperatively and the costs were EUR 5,100. Unstable fractures without neurological deficits were treated either nonoperatively, with an average of 29 hospitalization days and average cost of EUR 12,500 (86% of which represented hospitalization costs), or operatively with 24 hospitalization days and average cost of EUR 19,700 (48% of which represented hospitalization costs and 42% surgery costs). Unstable fractures with neurological deficits were usually operated (average costs EUR 31,900). INTERPRETATION: For all patients, the costs of hospitalization days were the main cost driver. Although the length of stay for patients with unstable fractures and without neurological deficit who were treated operatively was shorter than for patients treated nonoperatively, the total costs were higher due to the additional costs of surgery. Surgical treatment must therefore be shown to give a better outcome in order to outweigh the costs. Future research should focus on the cost-effectiveness of operative and nonoperative treatment of patients with unstable vertebral fractures who have no neurological deficits, and take indirect costs and quality of life into account. PMID- 16263614 TI - Outcome of operatively treated type-C injuries of the pelvic ring. AB - BACKGROUND: Internal fixation has become the preferred treatment for type-C pelvic ring injuries, but controversies persist regarding surgical approach and surgical technique. PATIENTS: We evaluated 101 consecutive patients with type C1 C3 pelvic ring injuries who had been treated with standardized reduction and internal fixation techniques. RESULTS: Our findings suggest a correlation between excellent reduction followed by sufficient fixation of the pelvic ring and functional outcome. Unsatisfactory reduction (displacement > 5 mm), failure of fixation, loss of reduction and a permanent lumbosacral plexus injury were the commonest reasons for an unsatisfactory functional result. All 40 patients with an associated lumbosacral plexus injury showed at least some evidence of neurological recovery. 14 underwent complete neurologic recovery. 8 had only sensory deficits and the remaining 18 also had motor deficits at the final followup. Complications were rare, but some of them were severe: loss of reduction in 8%, malunion in 10%, deep wound infection in 2%, and a lesion of the L5 nerve root in 1%. INTERPRETATION: Our results suggest that special attention should be paid to preoperative planning, reduction of the fracture, decompression of the nerve roots, and fixation of the most severe sacral fractures. Our results seem to favor internal fixation of displaced (> 10 mm) and unstable rami fractures and symphyseal disruptions in conjunction with posterior fixation, to achieve better stability of the whole pelvic ring. PMID- 16263615 TI - Cement-implant interface gaps explain the poor results of CMW3 for femoral stem fixation: A cadaver study of migration, fatigue and mantle morphology. AB - BACKGROUND: The Norwegian Arthroplasty Register reported that CMW3 cement performed poorly for femoral stem fixation. METHODS: We implanted collared, satin finished stems (Ra = 0.35 microm) into cadaver femora using CMW3 and with Simplex as control. Cement mantle function was quantified by stem migration after 300,000 cycles of "stair climbing". Cement cracks and interface gaps were quantified in transverse sections. RESULTS: The variances of the CMW3 migrations were substantially higher than for the control (p < 0.001): subsidence for CMW3: -32 (SD 42) microm, and for Simplex: -7 (SD 9) microm (p = 0.2); retroversion for CMW3: 0.60 degrees (SD 0.25), and for Simplex: 0.37 degrees (SD 0.04) (p = 0.08). Crack length-densities were similar. CMW3 had significantly more non-apposed stem/cement interface: 52% (SD 17) versus 33% (SD 8) (p = 0.04). Migrations could be predicted by the fraction of non-apposed stem/cement interface (retroversion: R(2)=0.80, p < 0.001; subsidence: R(2) = 0.46, p = 0.02) but not by cement cracks or non-apposed cement-bone interface. INTERPRETATION: We found that increased stem/cement non-apposition resulted in increased stem migration. Early migration is known to correlate with risk of revision. Thus, the higher stem-revision risk for CMW3 cement reported by the Norwegian Arthroplasty Register may have been due to inferior and variable stem/cement apposition. PMID- 16263616 TI - Oxford medial unicompartmental arthroplasty for focal spontaneous osteonecrosis of the knee. AB - BACKGROUND: Spontaneous osteonecrosis of the knee (SONK) is a distinct clinical condition occurring in patients without any associated risk factors. There is controversy as to the best method of treatment, and the available literature would suggest that patients with SONK have a worse outcome than those with primary osteoarthrosis when arthroplasty is performed. We assessed the outcome of medial unicompartmental knee arthroplasty (UKA) using the Oxford prosthesis for end-stage focal spontaneous osteonecrosis of the knee (SONK; Ahlback grades III and IV). PATIENTS AND METHODS: We assessed 29 knees (27 patients) with spontaneous osteonecrosis of the knee using the Oxford Knee Score. 26 knees had osteonecrosis of the medial femoral condyle and 3 had osteonecrosis of the medial tibial plateau. All had been operated on using the Oxford Medial Unicompartmental Knee Arthroplasty (UKA). This group was compared to a similar group (28 knees, 26 patients) who had undergone the same arthroplasty, but because of primary osteoarthrosis. Patients were matched for age, sex and time since operation. The mean length of follow-up was 5 (1-13) years. RESULTS: There were no implant failures in either group, but there was 1 death (from unrelated causes) 9 months after arthroplasty in the group with osteonecrosis. The mean Oxford Knee Score in the group with osteonecrosis was 38, and it was 40 in the group with osteoarthrosis. INTERPRETATION: Use of the Oxford Medial UKA for spontaneous focal osteonecrosis of the knee is reliable in the short to medium term, and gives results similar to those obtained when it is used for patients with primary osteoarthrosis. PMID- 16263617 TI - Trochleaplasty for patellar instability due to trochlear dysplasia: A minimum 2 year clinical and radiological follow-up of 19 knees. AB - BACKGROUND: Recurrent patellar dislocation may be associated with trochlear dysplasia. Trochleaplasty is a surgical procedure which strives to deepen the trochlear groove. We evaluated the clinical and radiological effect of trochleaplasty after a minimum follow-up of 2 years. PATIENTS AND METHODS: We examined 19 knees in 16 patients at a mean of 3 years after trochleaplasty. Postoperatively, a subjective questionnaire, a Kujala score, and tests for potential patellar redislocation and apprehension were evaluated. On radiographs we evaluated the preoperative and postoperative crossing sign, trochlear depth, trochlear bump, and patellar height. On CT scans, the pre- and postoperative tibial tuberosity to trochlear groove distance (TTTG) and the patellar inclination angle were measured. RESULTS: 16 of 19 knees improved subjectively. The Kujala score increased from 56 to 80 points at the latest follow-up. None of the patients sustained a redislocation. 5 patients had medial parapatellar tenderness, including 4 with persistent apprehension. Radiological signs of trochlear dysplasia were corrected. INTERPRETATION: Patellofemoral instability with underlying trochlear dysplasia can be treated successfully by trochleaplasty. PMID- 16263619 TI - An economic analysis of management strategies for closed and open grade I tibial shaft fractures. AB - BACKGROUND: Closed and open grade I (low-energy) tibial shaft fractures are a common and costly event, and the optimal management for such injuries remains uncertain. METHODS: We explored costs associated with treatment of low-energy tibial fractures with either casting, casting with therapeutic ultrasound, or intramedullary nailing (with and without reaming) by use of a decision tree. RESULTS: From a governmental perspective, the mean associated costs were USD 3,400 for operative management by reamed intramedullary nailing, USD 5,000 for operative management by non-reamed intramedullary nailing, USD 5,000 for casting, and USD 5,300 for casting with therapeutic ultrasound. With respect to the financial burden to society, the mean associated costs were USD 12,500 for reamed intramedullary nailing, USD 13,300 for casting with therapeutic ultrasound, USD 15,600 for operative management by non-reamed intramedullary nailing, and USD 17,300 for casting alone. INTERPRETATION: Our analysis suggests that, from an economic standpoint, reamed intramedullary nailing is the treatment of choice for closed and open grade I tibial shaft fractures. Considering financial burden to society, there is preliminary evidence that treatment of low-energy tibial fractures with therapeutic ultrasound and casting may also be an economically sound intervention. PMID- 16263618 TI - Operative treatment of primary patellar dislocation does not improve medium-term outcome: A 7-year follow-up report and risk analysis of 127 randomized patients. AB - BACKGROUND: The best treatment for primary patellar dislocation has been the subject of debate. Surgery has been recommended for all patients or for special subgroups to improve outcome. We have previously reported similar 2-year results after closed or open treatment. This report concerns 127 patients who were re evaluated by questionnaire at least 5 years after the primary onset. PATIENTS AND METHODS: At baseline, the patients were randomized regarding closed treatment (57) or individually adjusted proximal realignment operation (70). All patients were re-evaluated after a mean follow-up of 7 (6-9) years. RESULTS: The outcomes were similar: the patient's own overall opinion was excellent or good after closed treatment in 81% of cases and after operative treatment in 67%. Mean Kujala and Hughston VAS knee scores were 90 and 94, respectively, after closed treatment and 88 and 89 after operative treatment. The proportions of stable patellae were 30% and 36% for closed treatment and operative treatment, respectively. In a multivariate risk analysis, there was a correlation between a Kujala score of less than 90 and female sex (OR: 3.5; 95% CI: 1.4-9.0), loose bodies on radiographs (4.1; 1.2-15), and also an initial history of contralateral patellar instability (3.6; 0.9-15). There were 2 risk factors for recurrent instability: initial contralateral instability (4.9; 0.9-28) and young age (0.9; 0.8-1.0/year). Girls with open tibial apophysis had the worst prognosis for instability (88%; 95% CI: 77-98). INTERPRETATION: We do not recommend proximal realignment surgery for treatment of primary dislocation of the patella. PMID- 16263620 TI - Kinematics before and after reconstruction of the anterior syndesmosis of the ankle: A prospective radiostereometric and clinical study in 5 patients. AB - BACKGROUND: We have previously shown that patients with instability of the anterior syndesmosis benefit from an anatomical reconstruction. It is not known whether this is because of restored kinematics. METHODS: In a prospective study of 5 patients, we assessed clinical findings and tibiofibular kinematics, evaluated by radiostereometry, before and after reconstruction of a chronic syndesmotic injury. RESULTS: We found no statistically significant differences in tibiofibular kinematics before and after reconstruction. The kinematics of the fibula relative to the tibia during external rotation stress differed from that known in asymptomatic volunteers, but the differences were not typical enough to differentiate between patients and healthy subjects. Clinical examination and ankle scores, however, showed that all patients benefited from reconstruction of the anterior syndesmosis. INTERPRETATION: Radiostereometry is not an adequate technique to diagnose chronic syndesmotic instability or to demonstrate restoration of the kinematics of the ankle as a cause of the beneficial effect of anatomical reconstruction of the syndesmosis. PMID- 16263621 TI - Arthroscopically treated intraarticular osteoid osteoma in the ankle--a report of 3 cases. AB - We present 3 cases of arthroscopically treated intraarticular osteoid osteoma of the ankle. Emphasis is on the high index of suspicion for this lesion, the appropriate imaging modality and the efficacy of arthroscopic excision of these tumors with superficial location in the talus and tibial plafond. PMID- 16263622 TI - Did the use of crutches in an elderly patient activate a dormant subclavian lymphatic malformation?--a case report. PMID- 16263623 TI - Massive ectopic calcification of muscles in the lower leg following femoral artery injury-a case report. PMID- 16263627 TI - Growth and growth hormone in children born small for gestational age. AB - Although most children born small for gestational age catch up in growth by age 2 y, up to 14% remain more than 2 standard deviations below the mean for height. Recombinant growth hormone is approved by the US Food and Drug Administration and by the European Agency for Evaluation of Medicinal Products for the treatment of children born small for gestational age who fail to manifest catch-up growth by 2 y or 4 y, respectively. CONCLUSION: We conclude from clinical studies that growth hormone therapy can induce catch-up growth in these children. PMID- 16263628 TI - Phototherapy: old questions, new answers. AB - Despite its popularity and widespread use, there remains much more to be learned about phototherapy. CONCLUSION: There is continued debate on whether intermittent phototherapy is more effective than continuous phototherapy; if light of a longer wavelength is more effective than blue light phototherapy; and if phototherapy in the presence of bilirubin is toxic to normal or abnormal red blood cells. Three in vitro studies in this issue of the journal provide answers to some of these questions. PMID- 16263629 TI - Teenage asthma after severe infantile bronchiolitis or pneumonia. AB - OBJECTIVE: The purpose of the study was to evaluate asthma at >13 y of age in children with infantile bronchiolitis or pneumonia. METHODS: In 1981-1982, 127 children at <2 y of age were hospitalized for bronchiolitis (n = 81) or pneumonia (n = 46). Respiratory syncytial virus (RSV) infection, eosinophilia and markers of atopy were assessed and recorded on admission. At a median age of 14.9 y, atopic and asthmatic symptoms were screened by a written questionnaire in 98/127 (77%) study subjects. RESULTS: Asthma was present, according to two definitions, in 14% to 23% in the original bronchiolitis and in 12% to 15% in the original pneumonia group. The figures were 8% to 17% in the RSV infection and 16% to 23% in the non-RSV infection group. Early asthma-predictive factors were repeated wheezing, atopic dermatitis and elevated blood eosinophils. All but one of the teenage asthmatics had allergic rhinitis. CONCLUSION: An increased risk for asthma persists until the teenage period after bronchiolitis and pneumonia in infancy. Both early and later atopy were significant risk factors. The present study was unable to demonstrate the association between early RSV infection and teenage asthma. PMID- 16263630 TI - Bilirubin-induced cell death during continuous and intermittent phototherapy and in the dark. AB - AIM: To compare continuous and intermittent light exposure in the presence of bilirubin with respect to cellular damage. Furthermore, it was of interest to characterize the nature of cellular toxicity of bilirubin in the dark. METHOD: A murine lymphoma cell line, L5178Y-R (LY-R), was exposed to solutions of bilirubin (160 microM) supplemented with human serum albumin (200 microM) and irradiated with phototherapy light (Philips 20W/52) at a constant total dose of approximately 500 kJ/m2. The irradiation was given either as intermittent or continuous treatment with light of variable irradiance. The three lower irradiance levels were clinically relevant. Cells treated with bilirubin were also kept in the dark for various periods of time. Cell toxicity was determined by measuring apoptosis and necrosis. Apoptosis was measured by terminal deoxynucleotide transferase and propidium iodide staining assay, while trypan blue assay was used for detection of necrosis. RESULTS: There was no difference (n = 6, p > 0.05) between continuous and intermittent irradiation in the induction of early and late apoptotic cell death. Necrosis was more pronounced after intermittent treatment. Bilirubin dark toxicity was observed and classified as both apoptotic and necrotic. CONCLUSION: Continuous and intermittent light exposure caused the same degree of apoptotic cell death, while the cells underwent more necrotic death after intermittent exposure. Bilirubin was cytotoxic in the dark by both cell death mechanisms. PMID- 16263632 TI - Formation of photoproducts and cytotoxicity of bilirubin irradiated with turquoise and blue phototherapy light. AB - AIM: To compare a new turquoise ("green") fluorescent phototherapy lamp (490 nm) with a conventional blue phototherapy lamp (450 nm) with respect to cytotoxicity and photochemical effects of bilirubin. METHODS: Mouse lymphoma cells (L5178Y-R) in the presence of bilirubin solutions were exposed to phototherapy light. Occurrence of necrosis and apoptosis, reduction of mitotic index and inhibited cell growth was assayed by appropriate methods. The presence of bilirubin and its photoisomers was measured by high-pressure liquid chromatography analysis and absorption spectroscopy. RESULTS: At constant and equal light irradiances, the cytotoxic effects in the presence of bilirubin bound to human serum albumin showed that the green lamp caused significantly less necrosis (n = 4, p < 0.05) and less inhibition of cell multiplication (n = 3, p < 0.05) than the blue lamp. A slightly lower apoptotic fraction, although not statistically significant, was observed in cells exposed to the blue lamp. Photo-oxidation of bilirubin was more prominent with blue light irradiation. The photoequilibria between geometric isomers of bilirubin were different for the two lamps; more geometric photoisomers were formed by blue irradiation (n = 6, p < 0.05). The amounts of the most water-soluble isomers (presumably mainly lumirubin) were rather similar for the two lamps. CONCLUSION: The two lamps were similar in the formation of therapeutically relevant photoproducts, but the blue lamp showed potential in forming more photo-oxidation products and in causing more severe cellular damage in the presence of bilirubin. PMID- 16263633 TI - Doppler ultrasonographic detection of nutcracker syndrome in a young child with intussusception: a case report. AB - We report on a 2-y-old male with intussusception accompanying nutcracker syndrome detected by renal Doppler ultrasound. Renal Doppler ultrasound revealed a significant difference in the peak velocity between the hilar and aortomesenteric portions of the left renal vein. CONCLUSION: Renal Doppler ultrasound has been very useful and effective in detecting nutcracker syndrome in a young child. PMID- 16263631 TI - Effects of bilirubin and phototherapy on osmotic fragility and haematoporphyrin induced photohaemolysis of normal erythrocytes and spherocytes. AB - AIM: To study the effects of phototherapy on erythrocyte haemolysis in vitro and to determine possible differences in sensitivity to phototherapy between normal erythrocytes and spherocytes. METHODS: Erythrocytes from four normal healthy donors and two donors with hereditary spherocytosis were treated with bilirubin (160 microM) in the presence of human serum albumin in the molecular ratio bilirubin/albumin 0.8. Treated cells were maintained either in the dark or in blue light (450 nm, 8 mW/cm2, 30 min). The experimental light dose was comparable to 2 h of clinical phototherapy. The osmotic fragility of the treated cells was measured by scoring haemolysis in hypo-osmolar solutions (0.10-0.90% NaCl). The sensitivity to photohaemolysis of cells pre-treated with bilirubin (BR) and/or phototherapy was tested by exposing the cell suspensions to haematoporphyrin and UVA radiation. The delayed (18 h) photohaemolysis was measured by spectrophotometry. RESULTS: Osmotic fragility, expressed as percentage haemolysis, of normal erythrocytes was more than doubled in the presence of BR combined with phototherapy (n = 6, p < 0.05). In contrast, osmotic fragility of spherocytes was unaffected by either treatment (n = 8, p < 0.05). Increased photohaemolysis was seen in spherocytes treated with BR (n = 13, p < 0.05), phototherapy (n = 13, p < 0.05) and a combination of the two agents (n = 13, p < 0.05) compared with spherocytes without BR in the dark (n = 6). CONCLUSION: Bilirubin may make the plasma membrane of normal erythrocytes more fragile. Newborns with hereditary spherocytosis may be sensitive to phototherapy. PMID- 16263634 TI - Allergic colitis presenting within the first hours of premature life. AB - A prematurely born infant developed rectal blood loss several hours after birth, after his first formula feeding. Discontinuing the feeding resolved symptoms, but after resuming feeding rectal blood loss reappeared. There were no signs of necrotizing enterocolitis. Suspecting cow's milk allergy, the feeding was changed to a casein-based protein hydrolysate, without effect. Meanwhile, laboratory tests indicated cow's milk allergy. Symptoms only resolved after introducing an amino acid-based formula supporting a definite diagnosis of cow's milk-induced allergic colitis. This is the first description of a premature infant with symptoms of allergic colitis, appearing within hours after birth, suggestive of intrauterine sensitization. The exact mechanisms of sensitization remain obscure. CONCLUSION: Cow's milk-induced allergic colitis can occur after the first feed, even in a prematurely born neonate. This is most probably due to intrauterine sensitization, and should be included in the differential diagnosis of rectal blood loss. PMID- 16263635 TI - Discitis in toddlers: a case series and review. AB - Toddlers commonly present to medical services with non-specific symptoms. The presence of refusal to walk and nocturnal waking with crying should alert clinicians to the possibility of discitis. CONCLUSION: Features that may alert clinicians to the diagnosis of discitis in young children are reviewed. PMID- 16263638 TI - Randomization in clinical research. PMID- 16263639 TI - Caregiver burden, time spent caring and health status in the first 12 months following stroke. AB - OBJECTIVE: To quantify time caring, burden and health status in carers of stroke patients after discharge from rehabilitation; to identify the potentially modifiable sociodemographic and clinical characteristics associated with these outcomes. METHODS: Patients and carers prospectively interviewed 6 (n=71) and 12 (n=57) months after discharge. Relationships of carer and patient variables with burden, health status and time analysed by Gaussian and Poisson regression. RESULTS: Carers showed considerable burden at 6 and 12 months. Carers spent 4.6 and 3.6 hours per day assisting patients with daily activities at 6 and 12 months, respectively. Improved patient motor and cognitive function were associated with reductions of up to 20 minutes per day in time spent in daily activities. Better patient mental health and cognitive function were associated with better carer mental health. CONCLUSIONS: Potentially modifiable factors such as these may be able to be targeted by caregiver training, support and education programmes and outpatient therapy for patients. PMID- 16263640 TI - Factors related to recovery after mild traumatic brain injury. AB - PRIMARY OBJECTIVES: To study the variables that relate to outcome after mild traumatic brain injury (TBI). METHODS AND PROCEDURES: Sixty-seven adults with disappointing recoveries after mild TBI most occurring in a compensation or litigation context were studied with regard to pre-injury, neuro-trauma, physical, emotional and cognitive variables on outcome. Validity of physical, emotional and cognitive symptoms was controlled for. MAIN OUTCOMES AND RESULTS: Except for prior psychological traumatization, neither pre-injury, neuro-trauma or cognitive variables were related to outcome. Variables most consistently related to outcome were depression, pain and symptom invalidity on measures of response bias. These factors accounted for the majority of variance in outcome. CONCLUSIONS: In cases of poor recovery after mild TBI where compensation or litigation may be a factor, most of the variance in recovery seems to be explained by depression, pain and symptom invalidity, rather than by the injury variables themselves. PMID- 16263642 TI - Working memory and discourse production abilities following closed-head injury. AB - PRIMARY OBJECTIVES: This study investigated the relationship between working memory (WM) and narrative discourse production in individuals with closed head injury (CHI). It was hypothesized that those individuals with higher performance on tests of WM would demonstrate better performance on measures of discourse production. RESEARCH DESIGN: Correlation coefficients were calculated among five discourse measures from two story narratives and scores from three sub-tests of the Wechsler Memory Scale (WMS). METHODS AND PROCEDURES: Fifty-five individuals with moderate-to-severe CHI were studied. Participants included 16 females and 39 males ranging in age from 16-69. Narrative discourse samples were elicited from all participants under two conditions: story retelling and story generation. MAIN OUTCOMES AND RESULTS: The results revealed a number of modest, significant correlations (r=0.29-0.39, p<0.05) between sub-tests of the WMS and measures of discourse production. CONCLUSIONS: Results supported the hypothesis. Story elicitation task differences, limitations of using simple memory span tests as indices of WM and clinical implications of the relationship between WM and discourse production are discussed. PMID- 16263641 TI - Benefits of activity and virtual reality based balance exercise programmes for adults with traumatic brain injury: perceptions of participants and their caregivers. AB - OBJECTIVE: To explore multi-dimensional benefits of exercise participation perceived by adults with traumatic brain injury (TBI) and their caregivers. METHODS: Adults (n=27, aged 18-66) with moderate or severe TBI 6 months or more earlier participated in focus groups following 6 weeks of an activity-based (ABE) or a virtual reality (VR) delivered balance exercise programme. Family members and care providers participated in separate focus groups. Perceptions related to programme participation as well as balance confidence and lower extremity function were extracted from focus group verbatim and quantitative scales, respectively. OUTCOMES: Benefits in three domains, psychosocial, physical and programme, were identified from transcription and analyses of focus group verbatim. Improvements were noted in balance confidence and function in both groups. Substantially greater enthusiasm and knowledge was expressed by participants in the VR group and their caregivers. CONCLUSIONS: Both exercise programmes offered benefits in addition to improved balance. The VR participants had greater improvements on quantitative measures and provided more comments expressing enjoyment and improved confidence. Applications in terms of community reintegration and quality of life are discussed. PMID- 16263643 TI - Public attitudes towards survivors of brain injury. AB - PRIMARY OBJECTIVE: To explore the effects of religious identity, gender and socioeconomic status (SES) on public attitudes towards survivors of brain injury. RESEARCH DESIGN: An independent groups design was used to compare the attitudes of Northern Irish participants. METHODS AND PROCEDURES: The participants were asked to complete a modified form of the Community Attitudes to Mental Illness scale. The new questionnaire replaced the original scales' emphasis on mental illness with that of brain injury. Complete data was available for 179 participants for the religious identity and gender analysis and 124 for gender and SES. Analyses of variance were conducted on these variables. OUTCOMES AND RESULTS: Significant differences between male and female attitudes were found along with significant interactions between religious identity and gender and SES and gender. CONCLUSIONS: Religious, economic and gender-based divisions in society affect attitudes towards survivors of brain injury. PMID- 16263644 TI - Central auditory processing and social functioning following brain injury in children. AB - PRIMARY OBJECTIVE: To describe the percentage of children with central auditory processing disorders (CAPD) and examine the recovery patterns of social functional skills in children with and without CAPD admitted to inpatient rehabilitation following an acquired brain injury (ABI). RESEARCH DESIGN: Retrospective, descriptive. METHODS AND PROCEDURES: Repeated measures ANOVAs were used to examine overall differences in social functioning between groups, within groups and interaction effects for the Paediatric Evaluation of Disability Inventory (PEDI) Social Functional Skills and Caregiver Assistance scales for 31 children admitted to inpatient rehabilitation following ABI. MAIN OUTCOMES AND RESULTS: Significant overall effects were found in group differences and test occasions, but no significant interaction for either PEDI scale. Both groups showed significant intra-group changes between admission and discharge on both scales. CONCLUSIONS: Children with and without CAPD recovered social functional skills during inpatient rehabilitation. Children with CAPD were admitted and discharged with lower Social Function scores but demonstrated greater changes. The identification of CAPD during inpatient rehabilitation allows for appropriate discharge recommendations and realistic recovery expectations. PMID- 16263645 TI - The influence of victim characteristics on potential jurors' perceptions of brain damage in mild traumatic brain injury. AB - PRIMARY OBJECTIVE: To determine the influence of victim/plaintiff sex, occupation and intoxication status at the time of injury on potential jurors' judgement about the presence of brain damage in mild traumatic brain injury (MTBI). RESEARCH DESIGN: Survey. METHODS AND PROCEDURES: One of eight scenarios describing a MTBI from a motor vehicle accident was presented to 460 participants at a Department of Motor Vehicles. Victim sex, occupation (accountant or cafeteria worker) and alcohol intoxication status at the time of injury (sober or intoxicated) were manipulated across eight scenarios. Participants rated whether the victim's complaints at 6 months post-injury were the result of brain damage. MAIN OUTCOMES AND RESULTS: Ratings were influenced by victim occupation and intoxication status (chi2>5.3, p<0.03), but not the sex of the victim. CONCLUSIONS: The occupational and intoxication status of MTBI victims may influence potential jurors' decision about the presence of brain damage. PMID- 16263646 TI - Attentional deficits in concussion. AB - PRIMARY OBJECTIVE: The purpose of the present study was to examine deficits in the alerting, orienting and executive components of attention in individuals who have recently suffered a concussion. RESEARCH DESIGN: A group design was used in which the performance by individuals with concussion was compared to control subjects matched for age, height, weight and activity level. METHODS AND PROCEDURES: Participants completed the Attentional Network Test (ANT) that breaks down attention into alerting, orienting and executive components. Reaction time and response accuracy were the dependent variables. MAIN OUTCOMES AND RESULTS: It was found that only the orienting and executive components of attention were affected by concussion, whereas the alerting component was normal. Furthermore, participants with concussion required a significantly longer time than controls to initiate correct responses. CONCLUSIONS: These results suggest that the orienting and executive components of attention are most susceptible to the effects of concussion. PMID- 16263647 TI - Subjective well-being and quality of life following traumatic brain injury in adults: a long-term population-based follow-up. AB - PRIMARY OBJECTIVE: To assess subjective well-being and quality-of-life in nationally representative samples of patients at long intervals following traumatic brain injuries. METHODS AND PROCEDURES: Patients with either cranial fractures or cerebral lesions were identified in a national computer-based register of hospital admissions and random samples were selected among those who had suffered the injury at 5, 10 or 15 years prior to the follow-up. Postal questionnaires were sent to them covering quality of life, e.g. return to employment, family relations and current subjective well-being in terms of symptomatology, e.g. somatic complaints, cognitive dysfunction. A response rate of 76% was obtained, comprising 114 patients with cranial fracture and 126 with cerebral lesions. MAIN OUTCOMES AND RESULTS: The group with cerebral lesions had markedly poorer quality of life and subjective well-being than the group with cranial fractures and this did not vary across time. In both groups, the most common symptoms concerned cognition. Among the cerebral lesion group, quality of life outcome was fairly well predicted by severity of injury, but subjective well being was less well predicted. CONCLUSIONS: The negative consequences of traumatic cerebral lesions are marked and do not vary at long periods following injury. PMID- 16263648 TI - Psychosocial consequences of stroke: a long-term population-based follow-up. AB - PRIMARY OBJECTIVE: To investigate psychosocial status among nationally representative groups of stroke patients at long intervals post-stroke. METHODS AND PROCEDURES: From a Danish national register of hospitalizations, three representative groups of surviving patients were selected who had suffered a stroke 5, 10 and 15 years previously. A follow-up postal questionnaire was sent to them comprising items concerning symptomatology, functioning and social conditions, together with the Nottingham Health Profile (NHP). MAIN OUTCOMES AND RESULTS: Longer follow-up intervals were associated with younger age at stroke and better functioning at discharge. At follow-up, the majority of patients reported difficulties with attention, memory and emotional control, irrespective of follow-up interval. Return to employment, social relations and leisure activities were affected, but were comparatively better at longer follow-up intervals, as was self-rated functioning and several NHP symptom scales. However, multi-variate analyses suggest that these positive changes with time appear to be mediated by attrition related to age at stroke and discharge functioning rather than time since stroke itself. CONCLUSIONS: Symptomatology, functioning and social conditions remain affected and perhaps stagnant in long-term survivors of stroke. PMID- 16263649 TI - Pneumatosis intestinalis (PI) following severe traumatic brain injury. AB - PRIMARY OBJECTIVE: To report a case of pneumatosis intestinalis complicated with sorbitol containing medicines. RESEARCH DESIGN: Case report. METHODS AND PROCEDURES: Clinical course and literature review. EXPERIMENTAL INTERVENTIONS: Chart review. MAIN OUTCOMES AND RESULTS: Large amount of sorbital may cause a reversible gastrointestinal complication, pneumatosis intestinalis. CONCLUSIONS: In the assessment of enteral abnormalities, all components of medications, active and 'inactive', must be evaluated as possible aetiological factors of pneumatosis intestinalis. PMID- 16263650 TI - An unusual case of Dantrolene sodium-induced urinary retention in post-traumatic minimally responsive state. AB - INTRODUCTION: An unusual case of urinary retention is reported occurring during the inpatient rehabilitation of minimally responsive state (MRS) following severe traumatic brain injury (TBI). CLINICAL PICTURE: Urodynamic evaluation showed detrusor acontractility and subsequent management involved intermittent bladder catheterization and treatment of a single urinary tract infection. TREATMENT: Factors contributing to her bladder paralysis included severe immobility, tetraplegia and treatment of diffuse spasticity with oral Dantrolene sodium 250 mg per day. Constipation, diabetes and spinal cord injury were absent. In addition, she received unilateral partial sciatic neurolysis with 50% alcohol for severe knee flexor spasticity and intra-muscular Botulinum toxin A to both spastic upper limbs. OUTCOME: Bladder acontractility resolved completely when Dantrolene was reduced with subsequent achievement of a catheter-free status and small post-void residual volumes. Repeat urodynamics showed spontaneous detrusor contractions. CONCLUSION: A discussion of possible aetiologic factors for detrusor acontracility following TBI is presented including a brief review of the literature. PMID- 16263651 TI - Enhancement of the cell-killing effect of ultraviolet-C radiation by short-term exposure to a pulsed magnetic field. AB - PURPOSE: To investigate whether low frequency pulsed magnetic field (PMF) exposures produce alterations in the cell killing induced by ultraviolet C (UVC) radiation. MATERIALS AND METHODS: MCF-7 breast cancer cells of exponentially growing cultures were exposed to PMF (25 Hz, 0.75 mT) and UVC (from 6.6 J/m2 to 59.4 J/m2) in two different protocols: (a) cells were exposed to PMF for 30 min and then exposed to UVC at different doses; (b) cells were exposed to PMF for 30 min. After 15 min of the PMF exposure they were exposed simultaneously to PMF+different doses of UVC. After an additional time of 72 h of incubation, viability was measured by the neutral red stain cytotoxicity test. RESULTS: Both exposure protocols produced a significant decrease in the post UVC survival at 13.2 J/m2 and 19.8 J/m2, as compared to controls. The simultaneous exposition of PMF and UVC produced an additional increment in cell killing at 26.4 J/m2, being the greater effects obtained for this second exposure protocol. CONCLUSIONS: Results of the present study show that PMF in combination with UVC have the ability to augment the cell killing effects of UVC radiation. In addition, the effects appear to be greater when PMF and UVC are applied at the same time. PMID- 16263652 TI - Histological characteristics of cutaneous and thyroid mast cell populations in male rats exposed to power-frequency electromagnetic fields. AB - PURPOSE: The objective of this study was to determine whether mast cells (MC) in skin and thyroid gland, cutaneous nerve fibers and eosinophils are sensitive to the influence of electromagnetic fields (EMF). MATERIALS AND METHODS: The experiment was performed on two-month-old Wistar male rats, exposed to 50 Hz EMF (100-300 microT, 54-160 V/m) for 4 h a day, seven days a week during one month. After sacrifice, samples of skin and thyroid were processed for toluidine blue staining or indirect immunohistochemistry. The M42 grid placed in the ocular of a light microscope and a special microscopic frame placed in the ocular of a fluorescence microscope were used for stereological analysis. RESULTS: The numerical and volume density of intact type A MC in the thyroid of the exposed group was significantly higher compared to the control. A number of MC and immunoreactive nerve fibers were observed in the skin and of histamine immunoreactive MC in the thyroid of exposed animals. The differences in stereological data were not statistically significant by the Mann-Whitney test. CONCLUSIONS: The results indicate certain alterations of cutaneous and thyroid MC in rats exposed to EMF. However, the possible outcome of changes in the MC population under EMF influence on morphophysiological properties of other structures in skin and thyroid requires further investigation. PMID- 16263653 TI - No correlation between radiosensitivity or double-strand break repair capacity of normal fibroblasts and acute normal tissue reaction after radiotherapy of breast cancer patients. AB - The aim was to study the relationship between cellular radiosensitivity or double strand break (dsb) repair capacity of skin fibroblasts and the extent of acute reaction after radiotherapy for breast cancer. The study was performed with 25 breast cancer patients submitted to the radiotherapy unit of the Egyptian National Cancer Institute after conserving surgery. Dermal fibroblasts, established from skin biopsies, were used to determine the cellular radiosensitivity via colony assay and the capacity of dsb repair by constant field gel electrophoresis. Acute reactions were scored using the Radiation Therapy Oncology Group (RTOG) classification. The spectrum of acute reactions varied from grade 1 to 4, whereby most patients developed a grade 1 reaction after total doses ranging between 46 and 70 Gy. Skin fibroblasts showed a pronounced variation in both cellular radiosensitivity expressed as the mean inactivation dose (Dbar) (coefficient of variation, CV=25%) as well as in the number of residual dsb (CV=33%) with no significant correlation between these two endpoints (r2=0.20, p=0.14). Both parameters did not correlate with the extent of acute reaction of the respective patient. The data obtained indicate that the sensitivity of fibroblasts measured either by colony assay or by dsb repair capacity is not a major parameter determining the extent of acute reaction after radiotherapy of breast cancer patients. PMID- 16263654 TI - Adaptive response of a new radioresistant strain of Chlamydomonas reinhardtii and correlation with increased DNA double-strandbreak rejoining. AB - In order to study the relationship between radioresistance and the adaptive response, we aimed to produce a new strain of Chlamydomonas reinhardtii with characteristics of high radioresistance coupled with a protoplast structure typical for the genus, and the cell-wall-less phenotype to facilitate rapid cell lysis in DNA double-strand break (DSB) assays. The adaptive response of the new strain was investigated using clonogenic and DSB assays. Strain H-3 was derived by mating a radioresistant strain (AK-9-9) with the cell-wall-less mutant CW15 strain and selecting for radioresistance by clonogenic assay. The random amplification of polymorphic DNA (RAPD) molecular marker system was used to evaluate genetic polymorphisms between H-3 and other related C. reinhardtii strains. DSB were estimated using constant-field electrophoresis. Of several mutant strains tested, strain H-3 was shown to be most radioresistant on the basis of dose to give a 90% lethality (LD90) rate and dose to give a 99% lethality rate (LD99). In addition to its high radioresistance and thinner cell wall as compared with that of the other parental strain AK-9-9, H-3 also expressed a radiation-induced adaptive response measured by clonal survival when given a priming dose before a test dose. DSB were also rejoined more rapidly in cells exposed to a priming dose 4 h previously. It is concluded from split-dose experiments that the already highly radioresistant strain H-3 is further capable of 'over recovery' or adaptation to radiation exposure. Accelerated DSB rejoining in cells given a priming dose may underlie the cellular adaptive response in this organism. PMID- 16263655 TI - Fast He2+ ion irradiation of DNA loaded with platinum-containing molecules. AB - PURPOSE: The association of radiotherapy and chemotherapy is an attractive approach to improve the therapeutic index of the treatment of tumors. A lot of work has been devoted to investigate the effects of X-ray, gamma-ray and neutron irradiation of DNA or living cells loaded with different chemical compounds containing heavy atoms like platinum. No such studies exist presently when fast atomic ions are chosen as ionizing particles. In the present work, we investigate quantitatively the increase of DNA breaks in complexes of plasmid-DNA loaded with platinum atoms under irradiation by fast atomic He2+ ions. MATERIALS AND METHODS: DNA Plasmids (pBR322) are incubated in solutions containing different concentrations of terpyridine platinum (PtTC). In some preparations, dimethyl sulfoxide (DMSO), a free radical scavenger, has been added in order to investigate the role of the free radicals. The complexes of DNA plasmids loaded with high-Z atoms are irradiated under atmospheric conditions by He2+ ions at an energy of 143 MeV/amu and a linear energy transfert (LET) of 2.24 keV/microm. Analysis of DNA damage--single and double strand breaks--is made by electrophoresis on agarose gels. RESULTS: The results show a significant increase in DNA strand breaks when platinum is present, indicating a radiosensitization by the high Z atoms. The increase in DNA damages is attributed to inner-shell ionization of a platinum atom by secondary electrons emitted along the He2+ tracks followed by an Auger deexcitation, leading, thus, to a local amplification of the radiative effects close to the DNA. The contributions of scavengeable- solvant mediated--indirect effects and non-scavengeable effects (direct ionization) are quantitatively evaluated. CONCLUSION: Enhancement of DNA breaks in plasmids loaded with heavy atoms like platinum and irradiated by atomic ions are observed. This finding suggests an enhancement of cell death rate will occur under irradiation by atomic ions when the cells contain high-Z atoms located close to DNA due to the increase of the DNA breaks. PMID- 16263656 TI - The yield of radiation-induced chromosomal aberrations in first division human lymphocytes depends on the culture time. AB - PURPOSE: To investigate two long-held beliefs in radiation cytogenetics that were seemingly contradicted by reports that: (a) protracting gamma-ray exposures over 0.5 h halves the induced aberration yield compared with acute exposure, and (b) that induced aberration yields in guaranteed first in vitro division metaphases (M1) vary with culture time. MATERIALS AND METHODS: Replicate blood samples were exposed for 3 min to 3.0 Gy gamma-rays and standard phytohaemagglutinin stimulated lymphocyte cultures were harvested at 10 times ranging from 45-72 h. Forty-eight hour cultures were also made from blood exposed to 3.0 Gy for 30 min. Slides were differentially stained, combining the harlequin method with fluorescent in-situ hybridization (FISH) painting of chromosomes 2, 3 and 5. M1 metaphases were scored for 1- and 2-way translocations involving the painted chromosomes and all unstable aberrations in the full genomes. RESULTS: Dicentric and translocation yields from the 30 min exposure were approximately 10% lower than in 48 h cultures from cells exposed for 3 min, although this reduction is not significant. Dicentric aberration yields from the 3 min exposed cells cultured over the range 45-72 h remained constant up to 51 h then rose to a different constant value beyond 60 h. The increase at 60-70 h compared with the yield at 48 h was about 50%. A marginal increase at later times was also observed for translocations. CONCLUSION: The protracted exposure experiment produced results consistent with the G-function hypothesis that models the dose rate effect. Therefore the previous report of a marked departure from this model was not confirmed. The reports of aberration yields increasing with time of arrival at metaphase were confirmed. Possible explanations are discussed; the intercellular distributions of aberrations, or of doses to the cells or heterogeneous radiosensitivity of lymphocyte sub-populations. None alone seems sufficient quantitatively to explain the magnitude of the effect. The implications for biological dosimetry, which employs cultures times of approximately 48 h, are considered to be minor. PMID- 16263657 TI - Loss of mitochondrial membrane potential and caspase activation enhance apoptosis in irradiated K562 cells treated with herbimycin A. AB - PURPOSE: We previously reported that herbimycin A (HMA) alters the mode of cell death of K562 cells induced by radiation and enhanced their radiosensitivity. In the present study, we explored the apoptosis-inducing activity of HMA and the fundamental mechanism via which it regulates radiation-induced cell death. MATERIALS AND METHODS: Chronic myelogenous leukemia (CML) cell line K562 was used. For X-irradiation and drug treatment, cells were plated at approximately 2x10(5) cells/ml. Exponentially growing cells were treated with 10 Gy of X-ray using a 6-MeV X-ray machine at a dose rate of 200-300 cGy/min. The cells were treated with 0.25 microM HMA immediately after irradiation and HMA remained for the entire culture period. The modes of cell death were discriminated by morphological changes, analysis of cell cycle, analysis of the mitochondrial events, and the expression of apoptosis-related proteins. RESULTS: Our data demonstrates that radiation induced a significant time-dependent increase of cell death and failed to sustain a prolonged G2 arrest in K562 cells. Radiation induced cell death caused the accumulation of cyclinB1 and weak nuclear fragmentation, suggesting a mitotic catastrophe. This mitotic catastrophe was dependent upon the mitochondrial permeability transition pore (PTP) opening and was independent of caspase-3. In contrast, K562 cells treated with radiation and HMA had an accelerated cell death and induced a p53-independent apoptosis. This apoptotic pathway was dependent upon an initial hyperpolarization of the mitochondrial inner membrane, following the release of cytochrome c and subsequent caspase-3 activation. CONCLUSIONS: Two mechanisms of radiation-induced cell death in K562 cells, mitotic catastrophe and apoptosis, are regulated through distinct pathways, mitochondria and caspase-independent and -dependent, respectively. The findings of this study may provide new insights into improving the efficiency of radiotherapy in CML patients. PMID- 16263660 TI - How difficult it can be to deliver health care to refugees and to people in rural communities. PMID- 16263658 TI - Sensitivity to radiation-induced apoptosis and neuron loss declines rapidly in the postnatal mouse neocortex. AB - Therapeutic brain irradiation can cause progressive decline in cognitive function, particularly in children, but the reason for this effect is unclear. The study explored whether age-related differences in apoptotic sensitivity might contribute to the increased vulnerability of the young brain to radiation. Postnatal day 1 (P1) to P30 mice were treated with 0-16 Gy whole-body X irradiation. Apoptotic cells were identified and quantified up to 48 h later using the TdT-UTP nick end-labelling method (TUNEL) and immunohistochemistry for activated caspase-3. The number of neuron-specific nuclear protein (NeuN) positive and -negative cells were also counted to measure neuronal and non neuronal cell loss. Significantly greater TUNEL labelling occurred in the cortex of irradiated P1 animals relative to the other age groups, but there was no difference among the P7, P14 and P30 groups. Irradiation decreased the %NeuN positive cells in the mice irradiated on P1, whereas in P14 animals, irradiation led to an increase in the %NeuN-positive cells. These data demonstrate that neocortical neurons of very young mice are more susceptible to radiation-induced apoptosis. However, this sensitivity decreases rapidly after birth. By P14, acute cell loss due to radiation occurs primarily in non-neuronal populations. PMID- 16263661 TI - Refugee women's health: collaborative inquiry with refugee women in Rwanda. AB - A collaborative capacity building experience in a Rwandan refugee camp with refugee women from the Democratic Republic of Congo (DRC) is described in this article. In service to the American Refugee Committee, I taught 13 refugee women how to plan and facilitate focus group sessions with the larger community of refugee women. The facilitators then conducted 18 focus group sessions gathering data from 100 refugee women. Thematic results included the health implications of poverty, the struggle to survive, the overburden of daily work, ambivalence about family planning, and the lack of freedom to express themselves. PMID- 16263662 TI - Korean women living in the United States as student wives. AB - Korean students' wives who travel with their husbands to the United States while they pursue academic degrees often experience the extremes of assimilation into the host culture. The purpose of this study is to address the following questions: (a) What are the processes used by Korean women to adapt to life in the United States, and (b) what are the barriers they encounter, as wives of students and mothers of their children, in adapting to this new life? Fourteen Korean wives were interviewed. All interviews were completed in Korean and transcribed verbatim. The study reveals eight phases to the basic social process of adjusting to life in the United States: preconfronting, confronting, discovering, undergoing crisis, seeking, reorienting, reflecting, and reconfronting. These were observed to be dynamic and recurrent ongoing phases that are likely to repeat themselves. The study findings provide health care practitioners with a better understanding of Korean wives' adjustment to life in the United States and suggest the development of culturally appropriate interventions. PMID- 16263663 TI - Health status and access to health care of documented and undocumented immigrant Latino women. AB - Immigrant Latino women represent about one fifth of the total Latino population; however, data on health status and access to care for this population is limited. Using secondary data, we used a cross-sectional study to examine sociodemographic, migration, health status, and access to health care characteristics of immigrant documented and undocumented Latino women in North Texas. Undocumented women were less likely to report having health insurance and a regular health care provider, and reported lower education and income. These results support the need for providing immigrant women with health services such as health fairs, affordable health insurance programs, community health services, and increased opportunities for participation in federal and state programs. PMID- 16263664 TI - Competing knowledge claims in the provision of antenatal care: a qualitative study of traditional birth attendants in rural Zimbabwe. AB - In this article we examine the role of traditional birth attendants (TBAs) in their care of women during pregnancy and childbirth, and highlight their views on the training they receive and the application of knowledge they gain. We also discuss the ways women and men reason around the practices of TBAs. Data were generated using focus group discussions and individual interviews with TBAs, women, and men. The accounts of the TBAs, women, and men indicate that the women combined traditional and professional care, reasoning that in this way they get the different forms of assurance that each offers for the proper pregnancy outcome. The accounts, moreover, suggest that little of the knowledge gained from the training, including the referral of women at high risk, was implemented. One reason for this appears to be the failure to reflect on local knowledge and realities in TBA training. We conclude that any efforts or plans to incorporate the two systems of care should acknowledge local knowledge and realities. Only then can the aim of reducing maternal and infant morbidity and mortality be achieved. PMID- 16263666 TI - Penn Summit. PMID- 16263665 TI - From sunrise to sunset: an ethnography of rural Ecuadorian women's perceived health needs and resources. AB - In this ethnographic study, we describe the perceived health needs and resources of women in two rural Ecuadorian communities. Nineteen women participated in semistructured interviews. Women worked 3 to 30 hours outside the home each week but described their financial resources as insufficient. Most participants perceived their health as fair to poor. Four themes emerged related to the participants' health needs: (1) lack of money, (2) "it's better to be alone," (3) the physical burden of women's roles, and (4) suffering/self-sacrifice. Traditional medicine, local experts in traditional remedies, and nutrition emerged as resources to prevent illness. We believe our results offer valuable information in facilitating participatory health projects for social change. PMID- 16263668 TI - Impact of liberalization of protocols for the use of morphine sulfate in an urban emergency medical services system. AB - OBJECTIVE: To investigate the impact of liberalization of paramedic management protocols for the use of morphine sulfate (MS). METHODS: A retrospective database analysis tallied and categorized MS use into seven conditions during two intervals--six months before (control) and six months after (study) the protocol change. RESULTS: In the control interval, 760 of 34,020 (2.2%) patients received MS. In the study interval, 999 of 30,320 (3.3%) received the drug, a 50% relative increase in MS use. MS use dramatically increased in two assessment categories: other painful medical conditions (19.0% vs. 2.8% of transports, relative risk [RR] 6.8, 95% confidence interval [CI] 5.2-8.9) and nontraumatic abdominal pain (9.2% vs. 1.9% of transports, RR 4.8, 95% CI 3.3-6.9). CONCLUSION: Liberalization of pain management protocols resulted in an appreciable increase in the use of MS only in medical categories, predominantly abdominal pain, with no apparent safety or misuse issues. PMID- 16263669 TI - Automated external defibrillator deployment in high schools and senior centers. AB - BACKGROUND: Policymakers with limited funds have been forced to make difficult decisions regarding which sites merit automated external defibrillators (AEDs). Guidelines have recommended that the allocation of AEDs be based largely on the site-specific risk of sudden cardiac death (cardiac arrest), with devices preferentially located at high-risk venues. However, there are limited data on whether such a strategy is being followed. The authors surveyed low-risk (schools) and high-risk (senior centers) venues to assess the availability of AEDs. METHODS: A random sample of 200 high schools was identified; 12 were excluded, resulting in a final sample of 188. Each was contacted to determine the number of students, number of teachers, availability of AEDs, and number of cardiac arrests during the 2001-02 academic year. For comparison, 20 licensed senior centers were surveyed to assess the availability of AEDs. RESULTS: Among 147 schools that responded to the survey, the mean student enrollment was 1,117 and the mean number of teachers was 58. There were three reported cases of cardiac arrest (2 students, 1 teacher), resulting in an annual incidence of cardiac arrest in a school of 2.1% (95% confidence interval [CI] 0.4%-5.7%). Thirty-seven schools (25%; 95% CI 18%-33%) reported having AEDs and 35 (27%) intended to purchase them. In contrast, among 20 senior centers, AEDs were available at two (10%; 95% CI 1%-32%), and the and there were four reported cardiac arrests (annual incidence 20%). CONCLUSION: The availability of AEDs across different sites may not correspond directly to the risk of cardiac arrest at these sites. PMID- 16263670 TI - The efficacy of nebulized albuterol/ipratropium bromide versus albuterol alone in the prehospital treatment of suspected reactive airways disease. AB - OBJECTIVE: Ipratropium bromide has demonstrated efficacy when added to albuterol for the treatment of reactive airways disease (RAD). Its prehospital use has not been explored. METHODS: A before-and-after design was used. Prehospital and emergency department (ED) medical records were examined retrospectively six months before and six months after institution of a new protocol, which allowed the addition of ipratropium bromide to all nebulized treatments with albuterol. Primary outcome measures included: changes in vital signs (heart rate, respiratory rate, oxygen saturation), clinical improvement as assessed by paramedics, and admission rates. RESULTS: A total of 371 patients were included (n = 192 albuterol alone, n = 179 ipratropium/albuterol). There was no statistically significant difference between groups with regard to the change in heart rate, respiratory rate, or oxygen saturation. In addition, there was no difference in the proportion of patients with clinical improvement or deterioration as assessed by paramedics. There was no statistically significant difference in the admission rate from the ED except in the subgroup of patients using a metered-dose inhaler at the time of illness. Of note, more than one-third (133/371) of the patients were ultimately determined to have a diagnosis other than RAD, the majority of whom were diagnosed as having cardiac disease. CONCLUSIONS: The addition of ipratropium bromide to albuterol for the prehospital treatment of RAD does not appear to result in clinical outcome improvements. A substantial number of patients enrolled in the study were diagnosed as having cardiac disease. PMID- 16263671 TI - The effect of an 18-hour electrical power outage on an urban emergency medical services system. AB - OBJECTIVES: To describe the experience of an urban, commercial ambulance provider during the multistate August 2003 electrical power outage (EPO) and to identify how such an event can affect an emergency medical services (EMS) system. METHODS: Data were abstracted from all dispatch and prehospital medical records generated during the 18-hour EPO. An hour-by-hour comparison was made between the median number of hourly EMS dispatches during the month of August 2003 and the total number of hourly EMS dispatches during the EPO. Presenting complaints were categorized, and all calls related to the EPO were identified. RESULTS: During the first ten hours of the EPO, hourly call volume increased an average of 250% (range 50%-500%) as compared with all of August 2003. The greatest increase in call volume occurred during the first seven hours of the EPO. During the EPO, 16% (n = 13) of patients presented with a heat-related complaint precipitated by the failure of air conditioning. In the same period, 37% (n = 31) of patients presented with a respiratory-related complaint, 52% (n = 16) of which were precipitated by the failure of home respiratory equipment such as ventilators, nebulizers, and oxygen concentrators. During the EPO, the average time spent on scene with patients increased 63%, from 11 to 18 minutes, when compared with the time spent 16 hours immediately prior and 14 hours immediately after the EPO. Average response and transport times did not change. CONCLUSION: Loss of power can cause a dramatic but short-term increase in call volume and lead to a disaster-like situation. EMS resources may be conserved by proactively assisting patients with home medical devices before they develop emergent conditions. Such a patient population might benefit, for example, from home deliveries of portable oxygen tanks or from the creation of temporary facilities that provide a climate controlled environment and electrically powered medical devices. PMID- 16263672 TI - Prehospital cardiac arrest in a paramedic first-responder system using the Utstein style. AB - OBJECTIVES: To describe the characteristics of patients found to have cardiac arrest and to evaluate the characteristics predictive of survival after cardiac arrest in a paramedic first-responder model. METHODS: All patients who suffered out-of-hospital cardiac arrest in the city of Reading, Ohio, from January 1998 to December 2003 were recorded in the Utstein style. The number and incidence rate of witnessed arrests, initial rhythms, rate of bystander cardiopulmonary resuscitation (CPR), and 30-day mortality rate were retrospectively collected. Demographics, time to hospital, and response times were evaluated as predictors of survival. RESULTS: Of those patients initially found to be in cardiac arrest, 14.3% were discharged alive. Witnessed arrests were more likely to result in live discharge of the patient. Whether bystander CPR was performed was not found to affect survival, nor was initial rhythm, although no patients initially found in asystole were discharged alive. No demographic characteristics or response times were predictive of survival. CONCLUSION: The rates of survival in this paramedic first-response system are favorable compared with basic emergency medical technician first-response systems. Further study using direct comparison methodology is warranted to confirm these findings. PMID- 16263673 TI - Occupational injuries among emergency medical services personnel. AB - BACKGROUND: Emergency medical services (EMS) personnel treat 22 million patients a year in the United States, yet little is known of their injury risks. OBJECTIVES: To describe the epidemiology of occupational injuries among EMS personnel, calculate injury rates, and compare the findings with those for other occupational groups. METHODS: This was a retrospective review of injury records kept by two urban agencies. The agencies submitted all 617 case reports for three periods between January 1, 1998, and July 15, 2002. The agency personnel worked an estimated 2,829,906 hours during the study periods. Cases were coded according to U.S. Department of Labor (DOL) criteria. RESULTS: Four hundred eighty-nine cases met the DOL inclusion criteria. The overall injury rate was 34.6 per 100 full-time (FT) workers per year (95% confidence interval [CI] 31.5-37.6). "Sprains, strains, and tears" was the leading category of injury; the back was the body part most often injured. Of the 489 cases, 277 (57%) resulted in lost workdays, resulting in a rate of 19.6 (95% CI 17.3-21.9) per 100 FT workers; in comparison, the relative risks for EMS workers were 1.5 (95% CI 1.35-1.72) compared with firefighters, 5.8 (95% CI 5.12-6.49) compared with health services personnel, and 7.0 (95% CI 6.22-7.87) compared with the national average. CONCLUSIONS: The injury rates for EMS workers are higher than rates reported by DOL for any industry in 2000. Funding and additional research are critical to further defining the high risks to EMS workers and developing interventions to mitigate this serious problem. PMID- 16263674 TI - Comparison of crashes involving ambulances with those of similar-sized vehicles. AB - OBJECTIVE: To describe the characteristics and associated occupant injuries of motor vehicle collisions (MVCs) involving ambulances as compared with MVCs involving similar-sized vehicles. METHODS: Motor vehicle crash data in Pennsylvania from 1997-2001 were analyzed to compare the characteristics of crashes involving ambulances with those involving vehicles of a similar size. Crash demographics (e.g., location of crash, roadway conditions, intersection type) and associated injuries were examined and compared using chi-square tests and Fisher's exact test. RESULTS: 2,038 ambulance MVCs and 23,155 crashes involving similar-sized vehicles were identified. Weather and road surface conditions were similar, but ambulance MVCs occurred with increased frequency on evenings and weekends. Ambulances were more likely to be involved in four-way intersection crashes (43% vs. 23%, p = 0.001), angled collisions (45% vs. 29%, p = 0.001), and collisions at traffic signals (37% vs. 18%, p = 0.001). More people were involved in ambulance MVCs (p = 0.001), with 84% of ambulance MVCs involving three or more people and 33% involving five or more people. Injuries were reported in more ambulance MVCs (76% vs. 61%, p = 0.001). Pedestrian involvement was rare (< 5% in both groups). CONCLUSION: Ambulance crashes occur more frequently at intersections and traffic signals and involve more people and more injuries than those of similar-sized vehicles. PMID- 16263675 TI - Effectiveness of self-applied tourniquets in human volunteers. AB - BACKGROUND: Tourniquets are not commonly used in routine extremity trauma, but can be vital for hemorrhage control in austere conditions. OBJECTIVE: To determine the effectiveness in human volunteers of currently available self applied tourniquets for extremity hemorrhage. METHODS: Seven tourniquets were tested on the thigh for elimination of detectable distal pulse by Doppler auscultation at the popliteal artery (experiment I, n = 18 subjects). The tourniquets that were effective in >or=80% of subjects in experiment I were tested for effectiveness on the upper arm by auscultation at the radial artery (experiment II, n = 12 subjects). RESULTS: Three of the seven tourniquets were effective in all subjects in experiment I; a fourth tourniquet was effective in 88%. Three of the four successful devices were also 100% effective in experiment II; the fourth was effective in 75%. Failure of tourniquets to eliminate distal Doppler pulse signal was due to inadequate mechanical advantage for tightening, device failure (breakage), or intolerable pinching or circumferential pain prior to pulse elimination. The Combat Application Tourniquet (North American Rescue Products, Inc.), the Emergency & Military Tourniquet (Delfi Medical Innovations, Inc.), and the Special Operations Forces Tactical Tourniquet (Tactical Medical Solutions, LLC) were all found to be 100% effective in elimination of distal arterial pulse in both the arm and the leg in all subjects. CONCLUSION: Some commercially available tourniquets do not reliably occlude arterial blood flow and may not be successful in preventing extremity exsanguination in a trauma patient. Potential purchasers of such devices should bear this in mind when selecting a device for clinical use. PMID- 16263676 TI - Radial pulse character relationships to systolic blood pressure and trauma outcomes. AB - BACKGROUND: Patient measurements that do not require monitoring equipment may be the only way to evaluate casualties in austere conditions to determine treatment and transport priority. Objective. To test the hypothesis that palpable pulse characteristics in the radial artery would estimate systolic blood pressure (SBP) and predict outcome in trauma patients. METHODS: Data were analyzed from the medical records of 342 trauma patients ranging from 18 to 50 years of age. Prehospital data were collected by helicopter emergency medical personnel at the scene of the injury. Based on radial pulse character, patients were divided into normal (n = 313) and weak (n = 29) groups. Those whose medical records did not describe pulse characters were not considered. Differences in SBP, mortality, and medical interventions between the radial-pulse-character groups were evaluated. RESULTS: The SBP taken at the scene was a mean of 26 mm Hg lower in those patients with weak radial pulse characters (102 mm Hg versus 128 mm Hg). Similarly, the lowest mean SBPs recorded in the field between the normal- and weak-pulse-character groups were 112 mm Hg and 99 mm Hg, respectively. Patient mortality increased with weak pulse character such that the mortality rats were 3% for the normal-pulse-character group and 29% for the weak-pulse-character group (odds ratio = 15.2). CONCLUSIONS: These preliminary data suggest that a weak radial pulse may be an acceptable method for initial rapid evaluation of trauma patients. This simple and rapid method of pulse evaluation should be considered for the triage of trauma patients in field conditions with limited instrumentation. PMID- 16263677 TI - Validation of low-acuity emergency medical services dispatch codes. AB - BACKGROUND: Computer-aided dispatch systems are used to assess the severity of a 9-1-1 caller's complaint and then assign an appropriate level of emergency medical services (EMS) response. OBJECTIVE: To evaluate a group of low-acuity codes (defined as requiring advanced life support [ALS] intervention in fewer than 10% of cases) that has been derived and validated in one community. METHODS: All of the 9-1-1 medical calls assigned to these predetermined emergency medical dispatch codes between January 1, 2004, and July 1, 2004, were analyzed. ALS care was defined as receiving one or more of the following: pulse oximetry measurement, blood glucose measurement, cardiac defibrillation, administration of any medication, airway maneuvers, or the placement of an intravenous (IV) catheter. A more restrictive definition of ALS care (use of IV fluid bolus, medication administration, intubation, or defibrillation) was also calculated. RESULTS: A total of 1,799 calls were assigned low-acuity dispatch codes, and 1,597 met inclusion criteria. None of the 26 dispatch codes were found to be low acuity by the study definition. Fifty-six percent of these patients received ALS care. Placement of an IV-catheter was the ALS intervention used most frequently (45% of cases), followed by pulse oximetry measurement (32%), glucose measurement (22%), medication administration (11%), intubation (0.13%), and defibrillation (0%). The medication administered most frequent was morphine. When using the more restrictive definition of acuity, patients in 19 of the 28 categories received ALS intervention less than 10% of the time. Patients in the other seven categories were considered high-acuity 13% to 36% of the time. CONCLUSION: Dispatch codes that had previously been determined to be low-acuity were found not to be so in this community. The variation in clinical practice is likely explained by a more precautionary approach to care in this EMS system and the increased use of analgesics. This study demonstrates the need to define the optimal subset of prehospital patients who would benefit from these treatments. PMID- 16263678 TI - Outcome of travelers who refuse transport after emergency medical services evaluation at an international airport. AB - OBJECTIVE: To determine the short-term outcome of patients refusing transport after emergency medical services (EMS) evaluation at an international airport. METHODS: This was a prospective, descriptive, observational study of patients who refused transport after evaluation by Philadelphia Fire Department paramedics at Philadelphia International Airport from July 2003 through March 2004. Paramedics contacted a medical command physician (MCP), who recorded the patient's contact information. Three days later, one investigator attempted to contact the patient to administer a survey of the medical course in the three days following the initial encounter. RESULTS: Of 90 patients enrolled, 64 (71%) were reached in follow-up. Their average age was 45 years (range 10 months to 80 years); 41 (63%) were female. The most common presenting complaints were trauma-related (22 patients, 34%), neurologic (12, 19%), and gastrointestinal (7, 11%). The most common reasons for refusing transport were belief that their complaint was not serious (48, 75%) and fear they would miss a flight (34 patients, 53%). In the three days following the initial encounter, no patients recontacted 9-1-1, 16 patients (25%) had a recurrence of their initial complaints, and 32 patients (50%) saw or talked to a physician. There was one hospitalization but no deaths. Among patients lost to follow-up, no deaths of U.S. citizens were detected. CONCLUSIONS: Most patients who refused transport after EMS evaluation at an international airport had good short-term outcomes. These results may assist paramedics and MCPs to manage refusals in this setting and to allow patients to make informed decisions. PMID- 16263679 TI - Paramedic performance in calculating drug dosages following stressful scenarios in a human patient simulator. AB - BACKGROUND: Paramedics face many stressors in their work environment. Studies have shown that stress can have a negative effect on the psychological well-being of health professionals. However, there is little published research regarding the effects of stress on the cognitive skills necessary for optimal patient care. OBJECTIVES: The primary purpose of this study was to investigate the effects of acute stress on the emotional response and performance of paramedics. Furthermore, the authors explored whether a paramedic's level of training or years of experience would mediate the effects of stress on performance. METHODS: Paramedic performances in calculating drug dosages were compared in two stress conditions. In the low-stress condition, 30 paramedics calculated the drug dosages in a quiet classroom free of any stressor. In the high-stress condition, the same paramedics calculated comparable drug dosages immediately after working through a challenging scenario with a human patient simulator. RESULTS: The paramedics obtained lower accuracy scores in the high-stress condition than in the low-stress condition [43% (95% confidence interval [CI]: 36.9-49.2) vs. 58% (95% CI: 48.6-67.1), p < 0.01 based on univariate analysis]. Neither work experience nor level of training predicted the individual differences in the stress-induced performance decrements. CONCLUSION: These results suggest that the types of stressors encountered in clinical situations can increase medical errors, even in highly experienced individuals. These findings underline the need for more research to determine the mechanisms by which stress influences clinical performance, with the ultimate goal of targeting education or technologic interventions to those tasks, situations, and individuals most likely to benefit from such interventions. PMID- 16263680 TI - The EasyTube for airway management in emergencies. AB - BACKGROUND: The EasyTube (EzT) is a new sterile, disposable airway device approved by the European Union in February 2003 and by the U.S. Food and Drug Administration in January 2005. The two-lumen design of the EzT enables it to be used as an endotracheal tube or as a supraglottic emergency airway. OBJECTIVE: To report the preliminary experiences with the EzT airway device in prehospital and in-hospital emergency airway management procedures. METHODS: All airway management procedures involving the EzT were recorded for a period of 18 months. RESULTS: The EzT was successfully used to intubate 15 patients with unanticipated airway difficulties during either anesthesia induction or prehospital airway management. In all patients, the EzT was positioned successfully in the first attempt, within a median time of 31 seconds until start of ventilation. Effective supraglottic ventilation and oxygenation was achieved within 25 to 40 seconds. In three patients, the EzT needed one additional repositioning maneuver. On removal of the EzT, no blood was observed on the surface of the device, as a sign of absence of potential mucosal lesion. No injuries were observed in the mouth, pharynx, or esophagus. CONCLUSIONS: The first experiences with the use of the EzT are promising. In emergency airway management procedures presenting problems, the device successfully established sufficient ventilation and oxygenation. Further studies are needed to compare its value with those of other supraglottic devices. PMID- 16263681 TI - Transfer of intra-aortic balloon pump-dependent patients by paramedics. AB - This prospective case series describes the experience of a specially trained critical care paramedic transfer program that transports intra-aortic balloon pump (IABP)-dependent patients to definitive cardiac surgical care without additional medical escorts. Paramedics complete a standard ambulance patient care report and a quality management report for each IABP-dependent patient transported. All patient care reports were routinely screened for calls involving IABP-dependent patients to ensure all calls were captured. Demographic, patient care, adverse event, and transfer-related data were collected prospectively. The program manager and medical director independently examined all reports for quality of care and occurrence of adverse events. Missing data were obtained from paramedic crews and referring facilities. The IABP program transported 29 patients during its first 24 months. The mean patient age was 63.4 +/- 10.4 years, and the majority were male (25 of 29; 86.2%). The most common indications for IABP insertion were bridge to definitive surgical care (17; 58.6%) and cardiogenic shock (13; 44.8%). The mean out-of-hospital time was 39.9 +/- 26.1 minutes. There were 22 complications in 11 patients. Paramedics successfully managed all complications. There was no persistent life-threatening patient related complication or IABP-related malfunction, and no patient died during transport. This case series demonstrates that specially trained critical care paramedics can safely IABP-dependent patients to definitive cardiac surgical care without additional medical escorts. PMID- 16263682 TI - Severe hypothermia complicated by home heating fuel oil contamination. PMID- 16263683 TI - Advancing disaster epidemiology and response: developing a national disaster victim database. AB - BACKGROUND: The need to further disaster preparedness has resulted in a call for more comprehensive disaster research. Past disaster research has, for the most part, been limited by the inability to obtain complete medical data from victims of disasters. A national disaster-victim database (NDVD) can be developed that will facilitate collection and aggregation of disaster-victim medical data from health care facilities. Three aspects of the NDVD are discussed: DATA REQUIREMENTS: Medical records of disaster victims must be standardized before being uploaded from various databases into the NDVD. Existing data dictionaries provide formats in which data elements can become standardized. Once standardized, data sets from different facilities can be pooled and subjected to analyses. Database System. The three tiers of the NDVD system are: 1) medical data are collected at the point of care, 2) medical data are entered into databases and converted into a specific format, and 3) formatted data sets are uploaded to the NDVD. In order for this system to be viable, it must not add burden to health care workers; rather, it must benefit them and their facilities. REAL-TIME COLLECTION OF MEDICAL DATA: Capturing data on victims of a disaster, during a disaster, is ideal. This would allow for more victim data to be studied as well as for more accurate data to be collected. Technologic advancement has encouraged a real-time data-collection model in St. Louis that can act as a model for NDVD implementation. PMID- 16263684 TI - Policy position paper: National EMS organizations oppose establishing a U.S. Emergency Medical Services Administration within the Department of Homeland Security; support alternative approaches increasing federal support for EMS systems. PMID- 16263686 TI - Why isn't hypothermia after cardiac arrest the standard of care? PMID- 16263687 TI - Enantioselective and gender-dependent depletion of chlordane compounds from rat tissues. AB - Isomers and metabolites of the organochlorine pesticide chlordane persist in the environment and bioaccumulate in Arctic marine food webs. Rodent studies indicate that there are gender-related differences in trans-nonachlor and oxychlordane metabolism. Thus, comparative tissue depletion studies were undertaken in male and female rats exposed to trans-nonachlor, oxychlordane, or trans-chlordane at 2.5 mg/kg body weight/d by gavage for 28 d followed by two consecutive 28-d depletion periods. None of the test chemicals were overtly toxic at this dose, although increased liver weights in some groups were consistent with microsomal enzyme induction. The metabolite oxychlordane accumulated in tissues from rats exposed to trans-nonachlor and trans-chlordane. Trans-Nonachlor and oxychlordane residue levels were highest in tissues from female rats at each time point; however, trans-chlordane was completely eliminated from males and females by the end of the study. Body burden calculations showed no significant clearance of oxychlordane in females over 56 d postdosing, whereas males lost approximately half their oxychlordane body burden in the same period. For the chiral contaminants oxychlordane and trans-chlordane, tissues from male and female rats were selectively depleted of the (+)-enantiomer; however, there were gender related differences in enantiomer depletion patterns over time. In general, residue analyses confirmed that gender-related metabolic differences and contaminant structural properties, including chirality, influenced chlordane contaminant elimination from rat tissues. The study points to a need for similar knowledge of gender-related responses in humans in order to provide relevant dietary recommendations for populations exposed to chlordane-related contaminants in foods. PMID- 16263688 TI - The bipyridyl herbicide paraquat produces oxidative stress-mediated toxicity in human neuroblastoma SH-SY5Y cells: relevance to the dopaminergic pathogenesis. AB - Paraquat (PQ) is a cationic nonselective bipyridyl herbicide widely used to control weeds and grasses in agriculture. Epidemiologic studies indicate that exposure to pesticides can be a risk factor in the incidence of Parkinson's disease (PD). A strong correlation has been reported between exposure to paraquat and PD incidence in Canada, Taiwan, and the United States. This correlation is supported by animal studies showing that paraquat produces toxicity in dopaminergic neurons of the rat and mouse brain. However, it is unclear how paraquat triggers toxicity in dopaminergic neurons. Based on the prooxidant properties of paraquat, it was hypothesized that paraquat may induce oxidative stress-mediated toxicity in dopaminergic neurons. To explore this possibility, dopaminergic SH-SY5Y cells were treated with paraquat, and several biomarkers of oxidativestress were measured. First, a specific dopamine transporter inhibitor GBR12909 significantly protected SY5Y cells against the toxicity of paraquat, indicating that paraquat exerts its toxicity by a mechanism involving the dopamine transporter (DAT). Second, paraquat increased intracellular levels of reactive oxygen species (ROS), but decreased the levels of glutathione. Third, paraquat inhibited glutathione peroxidase activity, but did not affect glutathione reductase activity. On the other hand, paraquat increased GST activity by 24 h, after which GST activity returned to the control value at 48 h. Fourth, paraquat dissipated mitochondrial transmembrane potential (MTP). Fifth, paraquat produced increases of malondialdehyde (MDA) and protein carbonyls, as well as DNA fragmentation, indicating oxidative damage to major cellular components. Sixth, paraquat increased the protein level of heme oxygenase-1 (HO 1). Taken together, these findings verify our hypothesis that paraquat produces oxidative stress-mediated toxicity in SH-SY5Y cells. Thus, current findings suggest that paraquat may induce the pathogenesis of dopaminergic neurons through oxidative stress. PMID- 16263689 TI - Cholestyramine feeding lowers number of colonic apoptotic cells in rat. AB - Secondary bile acids that are formed in the colon by bacterial action have the potential property of eliciting pathological conditions. Apoptosis of mucosal epithelial cells is recognized as an adaptation that may counteract such pathologies. Cholestyramine, an anion exchange resin that sequesters bile salts in the gut, could decrease levels of secondary bile salt stress and thus conserve the potency of the protective action. Two groups of rats were studied: those fed 4% cholestyramine and those fed regular rat food. Rats were fed cholestyramine for 7, 14, 21, or 28 d. All animals were evaluated for cell death (apoptosis) using in situ TUNEL staining, and confirmed with single-stranded DNA (ssDNA). The effect of cholestyramine on the proliferating cell nuclear antigen (PCNA) in colonic crypt cells was also examined. Our data shows that animals fed cholestyramine for 28 d show evidence of a significant decrease in the levels of apoptotic cells in their large intestines, particularly goblet cells, when compared with the control animals and no change in cell proliferation. Thus, cholestyramine may serve as an alternative in attenuating apoptosis associated with inflammatory disorders that can result in significant enterocyte and goblet cell death. PMID- 16263690 TI - Non-coplanar PCB-mediated modulation of human leukocyte phagocytosis: a new mechanism for immunotoxicity. AB - Organochlorine (OC) contaminants, notably polychlorinated biphenyls (PCBs) and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), are ubiquitous in all ecosystems and found in the tissues of humans and wildlife. Although the immunotoxicity of coplanar, dioxinlike PCBs is well documented, the adverse effects exerted by non coplanar, non-dioxinlike PCBs have received little attention. Direct causal relationship between PCB and dioxin exposure and the observed detrimental effects on the immune system has yet to be fully established in humans. The immunomodulatory potential of toxic coplanar PCB 169 and TCDD and abundant non coplanar PCBs 138, 153, and 180 on human leukocyte phagocytosis, an important innate immune function that initiates the clearance of pathogens, was tested upon in vitro exposure. Mixture and concentration-response experiments demonstrated a suppression of phagocytosis by non-coplanar PCBs suggesting a previously unrecognized aryl hydrocarbon receptor (AhR)-independent pathway. Regression analysis revealed that reduction of phagocytosis was mostly explained by the non coplanar congeners. The effects on phagocytosis could not be accurately predicted by either the currently used toxic equivalence (TEQ) approach or the mouse model, thus undermining the use of the traditional models in the risk assessment for OC mixtures containing non-coplanar congeners. Our results are cause for concern as they suggest an AhR-independent pathway through which non-coplanar PCBs modulate phagocytosis, the immune system's first line of defense, possibly increasing the risk to developing infectious disease. PMID- 16263691 TI - Biological half-life and oxidative stress effects in mice with low-level, oral exposure to tritium. AB - Tritium ((3)H) may enter the environment from human activities, particularly at production, processing, or waste storage sites such as the Department of Energy's Savannah River Site, a former nuclear production facility in South Carolina. Understanding the dynamics and potential adverse effects of tritium in exposed organisms is critical to evaluating risks of tritium releases at such sites. Previous studies estimated the biological half-life of tritium in mice to be approximately 1.13 d; however, these laboratory studies were not conducted under environmentally realistic conditions. In this study, designed to be more representative of environmental exposure, mice were allowed to drink water containing tritium (activity about 300 Bq/ml) for a period of 2 wk. The induction of oxidative stress from tritium exposure was evaluated by comparing the activities of antioxidant enzymes (catalase, glutathione peroxidase, and superoxide dismutase) in exposed and control mice. From this experiment, the biological half-life of tritium was determined to be 2.26 +/- 0.04 d, almost double previous estimates. While positive controls (x-ray irradiated mice) showed responses in antioxidant enzyme activity, there was no indication of oxidative stress induction in mice exposed to tritium at this concentration. PMID- 16263692 TI - Mono-(3-carboxypropyl) phthalate, a metabolite of di-n-octyl phthalate. AB - Di-n-octyl phthalate (DnOP) is found as a component of mixed C6-C10 linear-chain phthalates used as plasticizers in various polyvinyl chloride applications, including flooring and carpet tiles. Following exposure and absorption, DnOP is metabolized to its hydrolytic monoester, mono-n-octyl phthalate (MnOP), and other oxidative products. The urinary levels of one of these oxidative metabolites, mono-(3-carboxypropyl) phthalate (MCPP), were about 560-fold higher than MnOP in Sprague-Dawley rats dosed with DnOP by gavage. Furthermore, MCPP was also found in the urine of rats dosed with di-isooctyl phthalate (DiOP), di-isononyl phthalate (DiNP), di-isodecyl phthalate (DiDP), di-(2-ethylhexyl) phthalate, and di-n-butyl phthalate (DBP), although at concentrations considerably lower than in rats given similar concentrations of DnOP. The comparatively much higher urinary concentrations of MCPP than of the hydrolytic monoesters of the high-molecular weight phthalates DiOP, DiNP, and DiDP in the exposed rats suggest that these monoesters may be poor biomarkers of exposure to their precursor phthalates and may explain the relatively low frequency of detection of these monoester metabolites in human populations. MCPP and MnOP were also measured in 267 human urine samples. The frequent detection and higher urinary concentrations of MCPP than MnOP suggest that exposure to DnOP might be higher than previously thought based on the measurements of MnOP alone. However, because MCPP is also a minor metabolite of DBP and other phthalates in rats, and the metabolism of phthalates in rodents and humans may differ, additional data on the absorption, distribution, metabolism, and elimination of MCPP are needed to completely understand the extent of human exposure to DnOP from the urinary concentrations of MCPP. PMID- 16263693 TI - Correlation of Asian dust storm events with daily clinic visits for allergic rhinitis in Taipei, Taiwan. AB - In spring, windblown dust storms originating in the deserts of Mongolia and China head toward Taipei city. These occurrences are known as Asian dust storm (ADS) events. The objective of this study was to assess the possible associations of ADS on the daily clinical visits for allergic rhinitis of residents in Taipei, Taiwan, during the period 1997-2001. Forty-nine dust storms were identified and classified as index days. Daily clinical visits for allergic rhinitis on the index days were compared with clinical visits on the comparison (non-ADS) days. Two non-ADS days for each index day, 7 d before the index day and 7 d after the index day, were selected. The effects of dust storms on clinic visits for allergic rhinitis were prominent 2 d after the event (19%). However, the association was not statistically significant. There may not have been enough power to detect associations resulting from the inadequate sample size of allergic rhinitis visits on ADS days. Further study is needed to clarify this association. PMID- 16263694 TI - Alprazolam abuse in Texas, 1998-2004. AB - Alprazolam (Xanax) is used in the treatment of anxiety, depression, and panic attacks, and is subject to abuse. The objective of this study was to describe the patterns of alprazolam abuse and drug identification (ID) calls received by several poison control centers. Cases were alprazolam calls received by 6 poison control centers during 1998-2004. Of 25,954 alprazolam calls received, 42% were drug ID calls and 51% were human exposure calls, of which 18% were abuse calls. The number of drug ID calls and the number of abuse calls both increased during the 7-yr period. Male patients accounted for 54% of abuse calls and females for 66% of nonabuse calls. Adolescent patients comprised 43% of abuse calls but only 12% of nonabuse calls. Although the majority of both types of human exposures occurred at the patient's own residence, abuse exposures were more likely than other exposures to occur at school (9% vs. 1%) and public areas (6% vs. 1%). While abuse calls were less likely than nonabuse calls to have no adverse clinical effects (19% vs. 23%), they were more likely to have minor medical outcomes (60% vs. 50%). Alprazolam abuse in Texas appears to be increasing. Alprazolam abusers are more likely to be male and often adolescent. Alprazolam abuse as compared to other exposures is more likely to occur outside of the person's home. Alprazolam abuse is more likely to involve some sort of adverse medical outcome. PMID- 16263695 TI - Relationship between radical generation by urban ambient particulate matter and pulmonary function of school children. AB - The mechanisms by which particulate matter (PM) produces adverse effects on the respiratory system, such as pulmonary dysfunction in children, are largely unknown. However, oxidative stress is thought to play an important role. Various chemical compounds in ambient particulate matter, including transition metals and aromatic organic compounds, may contribute to adverse effects through intrinsic generation of reactive oxygen species (ROS). It was hypothesized that ROS generation by PM, as determined through electron spin resonance (ESR) spectroscopy, may be negatively associated with pulmonary function in school children. PM(2.5), PM(10), and total suspended particulates (TSP) were sampled at the playgrounds of six elementary schools in the city of Maastricht, the Netherlands. All children (8-13 yr) from the six schools were asked to undergo spirometry. Multivariate linear regression models were constructed to evaluate associations between oxygen radical formation by PM and lung function. The radical-generating capacity per microgram PM correlated negatively to forced expiratory volume in 1 s (FEV(1)) and forced expiratory flow at 50% (FEF(50%)) of forced vital capacity (FVC). The data indicate that chemical features that contribute to intrinsic generation of ROS may be relevant for PM risk assessment. PMID- 16263696 TI - Androgen dependent mammary gland virilism in rats given the selective estrogen receptor modulator LY2066948 hydrochloride. AB - A selective estrogen receptor modulator (SERM) is a nonsteroidal compound with tissue specific estrogen receptor (ER) agonist or antagonist activities. In animals, SERMs may produce morphologic changes in hormonally-sensitive tissues like the mammary gland. Mammary glands from female rats given the SERM LY2066948 hydrochloride (LY2066948) for 1 month at >or= 175 mg/kg had intralobular ducts and alveoli lined by multiple layers of vacuolated, hypertrophied epithelial cells, resembling in part the morphology of the normal male rat mammary gland. We hypothesized that these SERM-mediated changes represented an androgen-dependent virilism of the female rat mammary gland. To test this hypothesis, the androgen receptor antagonist flutamide was co-administered with LY2066948 (175 mg/kg) to female rats for 1 month. Female rats given SERM alone had hyperandrogenemia and the duct and alveolar changes described here. Flutamide cotreatment did not affect serum androgen levels but completely blocked the SERM-mediated mammary gland change. In the mouse, a species that does not have the sex-specific differences in the mammary gland observed in the rat, SERM treatment resulted in hyperandrogenemia but did not alter mammary gland morphology. These studies demonstrate that LY2066948 produces species-specific, androgen-dependent mammary gland virilism in the female rat. PMID- 16263697 TI - Drug-induced protoporphyria in beagle dogs. AB - As part of regulatory safety testing program, a 13-week oral toxicity study with a new antipsychotic drug candidate was performed in beagle dogs. During this study, dark red/brown feces were recorded in treated dogs and increases in liver parameters (alanine aminotransferase, alkaline phosphatase, bilirubin) were measured biochemically. At the end of the study, livers of high-dose (50 mg/kg) animals were (mottled) dark brown, sometimes with pale foci. Histopathological examination of these livers showed dark globular pigment deposits in the hepatocellular cytoplasm and within the bile canaliculi. Varying numbers of inflammatory cell infiltrates were additionally present in association with the deposits. These pigment deposits showed birefringency with characteristic "Maltese Cross"-like structures under polarized light. Electronmicroscopy revealed the typical, so-called "sunburst" pattern with radiating double-lined crystalline structures. These morphologic characteristics strongly indicated at the presence of porphyrins, which was definitely confirmed biochemically. Published reports of drug-induced hepatic porphyria in dogs are rare. The possible underlying mechanism in the dog and man is discussed. PMID- 16263698 TI - Oncogenic signaling pathways activated in DMBA-induced mouse mammary tumors. AB - Only about 5% of human breast cancers can be attributed to inheritance of breast cancer susceptibility genes, while the balance are considered to be sporadic in origin. Breast cancer incidence varies with diet and other environmental influences, including carcinogen exposure. However, the effects of environmental carcinogens on cell growth control pathways are poorly understood. Here we have examined oncogenic signaling pathways that are activated in mammary tumors in mice treated with the prototypical polycyclic aromatic hydrocarbon (PAH) 7,12 dimethylbenz[a]anthracene (DMBA). In female FVB mice given 6 doses of 1 mg of DMBA by weekly gavage beginning at 5 weeks of age, all of the mice developed tumors by 34 weeks of age (median 20 weeks after beginning DMBA); 75% of the mice had mammary tumors. DMBA-induced mammary tumors exhibited elevated expression of the aryl hydrocarbon receptor (AhR), c-myc, cyclin D1, and hyperphosphorylated retinoblastoma (Rb) protein. Because of this, the activation of upstream regulatory pathways was assessed, and elements of the Wnt signaling pathway, the NF-kappa B pathway, and the prolyl isomerase Pin-1 were found to be frequently up regulated in the tumors when compared to normal mammary gland controls. These data suggest that environmental carcinogens can produce long-lasting alterations in growth and anti-apoptotic pathways, leading to mammary tumorigenesis. PMID- 16263699 TI - Elucidation of N-glycosylation sites on human platelet proteins: a glycoproteomic approach. AB - Among known platelet proteins, a prominent and functionally important group is represented by glycoprotein isoforms. They account e.g. for secretory proteins and plasma membrane receptors including integrins and glycoprotein VI as well as intracellular components of cytosol and organelles including storage proteins (multimerin 1 etc.). Although many of those proteins have been studied for some time with regard to their function, little attention has been paid with respect to their glycosylation sites. Here we report the analysis of N-glycosylation sites of human platelet proteins. For the enrichment of glycopeptides, lectin affinity chromatography as well as chemical trapping of protein bound oligosaccharides was used. Therefore, concanavalin A was used for specific interaction with carbohydrate species along with periodic acid oxidation and hydrazide bead trapping of glycosylated proteins. Derivatization by peptide:N glycosidase F yielded deglycosylated peptides, which provided the basis for the elucidation of proteins and their sites of modification. Using both methods in combination with nano-LC-ESI-MS/MS analysis 70 different glycosylation sites within 41 different proteins were identified. Comparison with the Swiss-Prot database established that the majority of these 70 sites have not been specifically determined by previous research projects. With this approach including hydrazide bead affinity trapping, the immunoglobulin receptor G6f, which is known to couple to the Ras-mitogen-activated protein kinase pathway in the immune system, was shown here for the first time to be present in human platelets. PMID- 16263700 TI - Low temperature treatment at the young microspore stage induces protein changes in rice anthers. AB - Male reproductive development in rice is very sensitive to various forms of environmental stresses including low temperature. A few days of cold treatment (<20 degrees C) at the young microspore stage induce severe pollen sterility and thus large grain yield reductions. To investigate this phenomenon, anther proteins at the early stages of microspore development, with or without cold treatment at 12 degrees C, were extracted, separated by two-dimensional gel electrophoresis, and compared. The cold-sensitive cultivar Doongara and the relatively cold-tolerant cultivar HSC55 were used. The abundance of 37 anther proteins was changed more than 2-fold after 1, 2, and 4 days of cold treatment in cv. Doongara. Among them, one protein was newly induced, 32 protein spots were up regulated, and four protein spots were down-regulated. Of these 37 protein spots, we identified two anther-specific proteins (putative lipid transfer protein and Osg6B) and a calreticulin that were down-regulated and a cystine synthase, a beta 6 subunit of the 20 S proteasome, an H protein of the glycine cleavage system, cytochrome c oxidase subunit VB, an osmotin protein homologue, a putative 6 phosphogluconolactonase, a putative adenylate kinase, a putative cysteine proteinase inhibitor, ribosomal protein S12E, a caffeoyl-CoA O-methyltransferase, and a monodehydroascorbate reductase that were up-regulated. Identification of these proteins is available upon request. Accumulation of these proteins did not vary greatly after cold treatment in panicles of cv. Doongara or in the anthers of the cv. HSC55. The newly induced protein named Oryza sativa cold-induced anther protein (OsCIA) was identified as an unknown protein. The OsCIA protein was detected in panicles, leaves, and seedling tissues under normal growth conditions. Quantitative real time RT-PCR analysis of OsCIA mRNA expression showed no significant change between low temperature-treated and untreated plants. A possible regulatory role for the newly induced protein is proposed. PMID- 16263701 TI - A novel subtractive antibody phage display method to discover disease markers. AB - Today's research demands fast identification of potential diagnostic and therapeutic targets. We describe a novel phage display strategy to identify disease-related proteins that are specifically expressed in a certain (diseased) tissue or cells. Phages displaying antibody fragments are selected on complex protein mixtures in a two-step manner combining subtractive selection in solution with further enrichment of specific phages on two-dimensional Western blots. Targets recognized by the resulting recombinant antibodies are immunoaffinity purified and identified by mass spectrometry. We used antibody fragment libraries from autoimmune patients to discover apoptosis-specific and disease-related targets. One of the three identified targets is the U1-70K protein, a marker for systemic lupus erythematosus overlap disease. Interestingly the epitope on U1-70K recognized by the selected recombinant antibody was shown to be apoptosis dependent, and such epitopes are believed to be involved in breaking tolerance to self-antigens. The other two proteins were identified as polypyrimidine tract binding protein-associated splicing factor (PSF)/nuclear RNA- and DNA-binding protein of 54 kDa (p54nrb) and heterogeneous ribonucleoprotein C. PMID- 16263702 TI - Proteomic studies of the Singapore grouper iridovirus. AB - The Singapore grouper iridovirus (SGIV) genome consists of a double-stranded circular DNA of 140,131 base pairs with 162 predicted open reading frames. Our earlier study using peptide mass fingerprints generated from MALDI-TOF MS led to the identification of 26 viral proteins. The present investigation aimed to achieve a more comprehensive and precise identification of the SGIV viral proteome by two workflows: one-dimensional gel electrophoresis (1-DE) separation followed by protein identification by MALDI-TOF/TOF MS/MS (1-DE-MALDI workflow) and shotgun proteomics in which the whole virus was digested by trypsin and the resulting peptides were separated by nano-LC and analyzed by MALDI-TOF/TOF MS/MS (LC-MALDI workflow). In total, 44 viral proteins were identified, 25 of which were reported for the first time. Fourteen proteins were uniquely identified by the 1-DE-MALDI workflow, whereas another 10 proteins were only identified by the LC-MALDI workflow with 20 proteins found by both approaches. Moreover 13 proteins were found to have acetylated N termini. Twenty-three proteins identified contain predicted transmembrane domains, accounting for 52.3% of the total proteins identified. RT-PCR confirmed the transcription products of all the identified viral proteins. A large number of proteins identified by both the 1-DE-MALDI and the LC-MALDI workflows from this study have significantly enhanced the coverage of the SGIV proteome. The SGIV proteome is at present the only iridoviral proteome that has been extensively characterized. Our results should provide further insights into the biology of SGIV and other iridoviruses. PMID- 16263703 TI - Induction of oxidative metabolism by mitochondrial frataxin inhibits cancer growth: Otto Warburg revisited. AB - More than 80 years ago Otto Warburg suggested that cancer might be caused by a decrease in mitochondrial energy metabolism paralleled by an increase in glycolytic flux. In later years, it was shown that cancer cells exhibit multiple alterations in mitochondrial content, structure, function, and activity. We have stably overexpressed the Friedreich ataxia-associated protein frataxin in several colon cancer cell lines. These cells have increased oxidative metabolism, as shown by concurrent increases in aconitase activity, mitochondrial membrane potential, cellular respiration, and ATP content. Consistent with Warburg's hypothesis, we found that frataxin-overexpressing cells also have decreased growth rates and increased population doubling times, show inhibited colony formation capacity in soft agar assays, and exhibit a reduced capacity for tumor formation when injected into nude mice. Furthermore, overexpression of frataxin leads to an increased phosphorylation of the tumor suppressor p38 mitogen activated protein kinase, as well as decreased phosphorylation of extracellular signal-regulated kinase. Taken together, these results support the view that an increase in oxidative metabolism induced by mitochondrial frataxin may inhibit cancer growth in mammals. PMID- 16263704 TI - Outer chain N-glycans are required for cell wall integrity and virulence of Candida albicans. AB - The outer layer of the Candida albicans cell wall is enriched in highly glycosylated mannoproteins that are the immediate point of contact with the host and strongly influence the host-fungal interaction. N-Glycans are the major form of mannoprotein modification and consist of a core structure, common to all eukaryotes, that is further elaborated in the Golgi to form the highly branched outer chain that is characteristic of fungi. In yeasts, outer chain branching is initiated by the action of the alpha1,6-mannosyltransferase Och1p; therefore, we disrupted the C. albicans OCH1 homolog to determine the importance of outer chain N-glycans on the host-fungal interaction. Loss of CaOCH1 resulted in a temperature-sensitive growth defect and cellular aggregation. Outer chain elongation of N-glycans was absent in the null mutant, demonstrated by the lack of the alpha1,6-linked polymannose backbone and the underglycosylation of N acetylglucosaminidase. A null mutant lacking OCH1 was hypersensitive to a range of cell wall perturbing agents and had a constitutively activated cell wall integrity pathway. These mutants had near normal growth rates in vitro but were attenuated in virulence in a murine model of systemic infection. However, tissue burdens for the Caoch1delta null mutant were similar to control strains with normal N-glycosylation, suggesting the host-fungal interaction was altered such that high burdens were tolerated. This demonstrates the importance of N-glycan outer chain epitopes to the host-fungal interaction and virulence. PMID- 16263705 TI - Characterization of rapidly degraded polypeptides in mammalian cells reveals a novel layer of nascent protein quality control. AB - Approximately 30% of polypeptides synthesized by mammalian cells are degraded with a half-life of <10 min by proteasomes. These rapidly degraded polypeptides (RDPs) constitute the bulk of proteasome substrates and are the principal source of viral and self-peptide ligands for major histocompatibility complex class I molecules. Here we provide evidence that approximately 75% of RDPs are degraded by the standard ubiquitin 26 S proteasome system and that their degradation is regulated by modulating Hsc70 activity in cells. Surprisingly, the remaining approximately 25% of RDPs are degraded without ubiquitylation by 20 S proteasomes independently of 19 S regulators and in a manner that is largely unaffected by modulating Hsc70 activity. This latter pathway is utilized for generating an antigenic peptide from viral-defective ribosomal products. The dichotomy in the behavior of RDPs points to a novel quality control level for nascent proteins that is independent of the well established Hsc70-ubiquitin 26 S proteasome pathway. PMID- 16263706 TI - Mediator as a general transcription factor. AB - Others have shown that yeast strains bearing a ts mutation in the Srb4 subunit of Mediator cease transcription of all mRNA at the restrictive temperature, in a manner virtually indistinguishable from a strain bearing a ts mutation in the largest subunit of RNA polymerase II. We find that srb4ts Mediator is defective for the stimulation of basal RNA polymerase II transcription at the restrictive temperature in vitro. Taken together, these findings lead to the suggestion that Mediator is required for basal RNA polymerase II transcription in vivo. On this basis, Mediator is identified as a general transcription factor, comparable in importance to RNA polymerase II and other general factors for the initiation of transcription. The possibility that Mediator serves as an anti-inhibitor, opposing the effects of global negative regulators, is largely excluded. PMID- 16263707 TI - A quantitative approach to catabolite repression in Escherichia coli. AB - A dynamic mathematical model was developed to describe the uptake of various carbohydrates (glucose, lactose, glycerol, sucrose, and galactose) in Escherichia coli. For validation a number of isogenic strains with defined mutations were used. By considering metabolic reactions as well as signal transduction processes influencing the relevant pathways, we were able to describe quantitatively the phenomenon of catabolite repression in E. coli. We verified model predictions by measuring time courses of several extra- and intracellular components such as glycolytic intermediates, EII-ACrr phosphorylation level, both LacZ and PtsG concentrations, and total cAMP concentrations under various growth conditions. The entire data base consists of 18 experiments performed with nine different strains. The model describes the expression of 17 key enzymes, 38 enzymatic reactions, and the dynamic behavior of more than 50 metabolites. The different phenomena affecting the phosphorylation level of EIIACrr, the key regulation molecule for inducer exclusion and catabolite repression in enteric bacteria, can now be explained quantitatively. PMID- 16263708 TI - Glycosphingolipid-facilitated membrane insertion and internalization of cobra cardiotoxin. The sulfatide.cardiotoxin complex structure in a membrane-like environment suggests a lipid-dependent cell-penetrating mechanism for membrane binding polypeptides. AB - Cobra cardiotoxins, a family of basic polypeptides having lipid- and heparin binding capacities similar to the cell-penetrating peptides, induce severe tissue necrosis and systolic heart arrest in snakebite victims. Whereas cardiotoxins are specifically retained on the cell surface via heparan sulfate-mediated processes, their lipid binding ability appears to be responsible, at least in part, for cardiotoxin-induced membrane leakage and cell death. Although the exact role of lipids involved in toxin-mediated cytotoxicity remains largely unknown, monoclonal anti-sulfatide antibody O4 has recently been shown to inhibit the action of CTX A3, the major cardiotoxin from Taiwan cobra venom, on cardiomyocytes by preventing cardiotoxin-induced membrane leakage and CTX A3 internalization into mitochondria. Here, we show that anti-sulfatide acts by blocking the binding of CTX A3 to the sulfatides in the plasma membrane to prevent sulfatide-dependent CTX A3 membrane pore formation and internalization. We also describe the crystal structure of a CTX A3-sulfatide complex in a membrane-like environment at 2.3 angstroms resolution. The unexpected orientation of the sulfatide fatty chains in the structure allows prediction of the mode of toxin insertion into the plasma membrane. CTX A3 recognizes both the headgroup and the ceramide interfacial region of sulfatide to induce a lipid conformational change that may play a key role in CTX A3 oligomerization and cellular internalization. This proposed lipid-mediated toxin translocation mechanism may also shed light on the cellular uptake mechanism of the amphiphilic cell penetrating peptides known to involve multiple internalization pathways. PMID- 16263709 TI - A Thr357 to Ser polymorphism in homozygous and compound heterozygous subjects causes absent or reduced P2X7 function and impairs ATP-induced mycobacterial killing by macrophages. AB - The P2X(7) receptor is a ligand-gated cation channel that is highly expressed on mononuclear leukocytes and that mediates ATP-induced apoptosis and killing of intracellular pathogens. There is a wide variation in P2X(7) receptor function between subjects, explained in part by four loss-of-function polymorphisms (R307Q, E496A, I568N, and a 5'-intronic splice site polymorphism), as well as rare mutations. In this study, we report the allele frequencies of 11 non synonymous P2X(7) polymorphisms and describe a fifth loss-of-function polymorphism in the gene (1096C --> G), which changes Thr(357) to Ser (T357S) with an allele frequency of 0.08 in the Caucasian population. P2X(7) function was measured by ATP-induced ethidium(+) influx into peripheral blood lymphocytes and monocytes and, when compared with wild-type subjects, was reduced to 10-65% in heterozygotes, 1-18% in homozygotes, and 0-10% in compound heterozygotes carrying T357S and a second loss-of-function polymorphism. Overexpression of the T357S mutant P2X(7) in either HEK-293 cells or Xenopus oocytes gave P2X(7) function of approximately 50% that of wild-type constructs. Differentiation of monocytes to macrophages, which also up-regulates P2X(7), restored P2X(7) function to near normal in cells heterozygous for T357S and to a value 50-65% of wild-type in cells homozygous for T357S or compound heterozygous for T357S/E496A. However, macrophages from subjects that are compound heterozygous for either T357S/R307Q or T357S/stop codon had near-to-absent P2X(7) function. These functional deficits induced by T357S were paralleled by impaired ATP-induced apoptosis and mycobacteria killing in macrophages from these subjects. Lymphocytes, monocytes, and macrophages from subjects homozygous for T357S or compound heterozygous for T357S and a second loss-of-function allele have reduced or absent P2X(7) receptor function. PMID- 16263710 TI - Rat long chain acyl-CoA synthetase 5 increases fatty acid uptake and partitioning to cellular triacylglycerol in McArdle-RH7777 cells. AB - Long chain acyl-CoA synthetase (ACSL) catalyzes the initial step in long chain fatty acid metabolism. Of the five mammalian ACSL isoforms cloned and characterized, ACSL5 is the only isoform found to be located, in part, on mitochondria and thus was hypothesized to be involved in fatty acid oxidation. To elucidate the specific roles of ACSL5 in fatty acid metabolism, we used adenoviral-mediated overexpression of ACSL5 (Ad-ACSL5) in rat hepatoma McArdle RH7777 cells. Confocal microscopy revealed that Ad-ACSL5 colocalized to both mitochondria and endoplasmic reticulum. When compared with cells infected with Ad GFP, Ad-ACSL5-infected cells at 24 h after infection had 2-fold higher acyl-CoA synthetase activities and 30% higher rates of fatty acid uptake when incubated with 500 microM [1-(14)C]oleic acid. Metabolism of [1-(14)C]oleic acid to cellular triacylglycerol (TAG) increased 42% in Ad-ACSL5-infected cells, but when compared with control cells, metabolism to acid-soluble metabolites, phospholipids, and medium TAG did not differ substantially. The incorporation of [1-(14)C]oleate and [1,2,3-(3)H]glycerol into TAG was similar in Ad-ACSL5 infected cells, thus indicating that Ad-ACSL5 increased TAG synthesis through both de novo and reacylation pathways. However, [1-(14)C]acetic acid incorporation into cellular lipids showed that, when compared with control cells, Ad-ACSL5-infected cells did not increase the metabolism of fatty acids that were derived from de novo synthesis. These results suggest that uptake of fatty acids into cells is regulated by metabolism and that overexpressed ACSL5 partitions exogenously derived fatty acids toward TAG synthesis and storage. PMID- 16263711 TI - Bcl-2 and Mn-SOD antisense oligodeoxynucleotides and a glutamine-enriched diet facilitate elimination of highly resistant B16 melanoma cells by tumor necrosis factor-alpha and chemotherapy. AB - Mitochondrial glutathione (mtGSH) depletion increases sensitivity of Bcl-2 overexpressing B16 melanoma (B16M)-F10 cells (high metastatic potential) to tumor necrosis factor-alpha (TNF-alpha)-induced oxidative stress and death in vitro. In vivo, mtGSH depletion in B16M-F10 cells was achieved by feeding mice (where the B16M-F10 grew as a solid tumor in the footpad) with an L-glutamine (L-Gln) enriched diet, which promoted in the tumor cells an increase in glutaminase activity, accumulation of cytosolic L-glutamate, and competitive inhibition of GSH transport into mitochondria. L-Gln-adapted B16M-F10 cells, isolated using anti-Met-72 monoclonal antibodies and flow cytometry-coupled cell sorting, were injected into the portal vein to produce hepatic metastases. In l-Gln-adapted invasive (iB16M-Gln+) cells, isolated from the liver by the same methodology and treated with TNF-alpha and an antisense Bcl-2 oligodeoxynucleotide, viability decreased to approximately 12%. iB16M-Gln+ cell death associated with increased generation of O2*- and H2O2, opening of the mitochondrial permeability transition pore complex, and release of proapoptotic molecular signals. Activation of cell death mechanisms was prevented by GSH ester-induced mtGSH replenishment. The oxidative stress-resistant survivors showed an adaptive response that includes overexpression of manganese-containing superoxide dismutase (Mn-SOD) and catalase activities. By treating iB16M-Gln+ cells with a double anti- antisense therapy (Bcl-2 and SOD2 antisense oligodeoxynucleotides) and TNF-alpha, metastatic cell survival decreased to approximately 1%. Chemotherapy (taxol plus daunorubicin) easily removed this minimum percentage of survivors. This contribution identifies critical molecules that can be sequentially targeted to facilitate elimination of highly resistant metastatic cells. PMID- 16263713 TI - Mycobacterium tuberculosis lipoarabinomannan-mediated IRAK-M induction negatively regulates Toll-like receptor-dependent interleukin-12 p40 production in macrophages. AB - Mannose-capped lipoarabinomannans (Man-LAMs) are members of the repertoire of Mycobacterium tuberculosis modulins that the bacillus uses to subvert the host innate immune response. Interleukin-12 (IL-12) production is critical for mounting an effective immune response by the host against M. tuberculosis. We demonstrate that Man-LAM inhibits IL-12 p40 production mediated by subsequent challenge with lipopolysaccharide (LPS). Man-LAM inhibits LPS-induced IL-12 p40 expression in an IL-10-independent manner. It attenuates LPS-induced NF-kappaB driven luciferase gene expression, suggesting that its effects are likely directly related to inhibition of NF-kappaB. This is probably because of dampening of the Toll-like receptor signaling. Man-LAM inhibits IL-1 receptor associated kinase (IRAK)-TRAF6 interaction as well as IkappaB-alpha phosphorylation. It directly attenuates nuclear translocation and DNA binding of c-Rel and p50. Man-LAM exerts these effects by inducing the expression of Irak-M, a negative regulator of TLR signaling. Knockdown of Irak-M expression by RNA interference reinstates LPS-induced IL-12 production in Man-LAM-pretreated cells. The fact that Irak-M expression could be elicited by yeast mannan suggested that ligation of the mannose receptor by the mannooligosaccharide caps of LAM was the probable trigger for IRAK-M induction. PMID- 16263712 TI - Cathepsin D and H2O2 stimulate degradation of thioredoxin-1: implication for endothelial cell apoptosis. AB - Cathepsin D (CatD) is a lysosomal aspartic proteinase and plays an important role in the degradation of proteins and in apoptotic processes induced by oxidative stress, cytokines, and aging. All of these stimuli are potent inducers of endothelial cell apoptosis. Therefore, we investigated the role of CatD in endothelial cell apoptosis and determined the underlying mechanisms. Incubation with 100-500 microm H2O2 for 12 h induced apoptosis in endothelial cells. To determine a role for CatD, we co-incubated endothelial cells with the CatD inhibitor pepstatin A. Pepstatin A as well as genetic knock down of CatD abolished H2O2-induced apoptosis. In contrast, overexpression of CatD wild type but not a catalytically inactive mutant of CatD (CatDD295N) induced apoptosis under basal conditions. To gain insights into the underlying mechanisms, we investigated the effect of CatD on reactive oxygen species (ROS) formation. Indeed, knocking down CatD expression reduced H2O2-induced ROS formation and apoptosis. The major redox regulator in endothelial cells is thioredoxin-1 (Trx), which plays a crucial role in apoptosis inhibition. Thus, we hypothesized that CatD may alter Trx protein levels and thereby promote formation of ROS and apoptosis. Incubation with 100 microm H2O2 for 6 h decreased Trx protein levels, whereas Trx mRNA was not altered. H2O2-induced Trx degradation was inhibited by pepstatin A and genetic knock down of CatD but not by other protease inhibitors. Incubation of unstimulated cell lysates with recombinant CatD significantly reduced Trx protein levels in vitro, which was completely blocked by pepstatin A pre-incubation. Overexpression of CatD reduced Trx protein in cells. Moreover, H2O2 incubation led to a translocation of Trx to the lysosomes prior to the induction of apoptosis. Taken together, CatD induces apoptosis via degradation of Trx protein, which is an essential anti-apoptotic and reactive oxygen species scavenging protein in endothelial cells. PMID- 16263714 TI - Polyamine transport by mammalian cells and mitochondria: role of antizyme and glycosaminoglycans. AB - The role of antizyme (AZ) and glycosaminoglycans in polyamine uptake by mammalian cells and mitochondria was examined using NIH3T3 and FM3A cells and rat liver mitochondria. AZ is synthesized as two isoforms (29 and 24.5 kDa) due to the existence of two initiation codon AUGs in the AZ mRNA. Most AZ existed as the 24.5-kDa form translatable from the second AUG, but a portion of the 29-kDa AZ from the first AUG was associated with mitochondria because of the presence of a mitochondrial targeting signal between the first and the second methionine. The predominance of the 24.5-kDa isoform was mainly due to the presence of spermidine and a favorable sequence context (Kozak sequence) at the second initiation codon AUG. Spermine uptake by NIH3T3 cells was inhibited by both 29- and 24.5-kDa AZs, but uptake by rat liver mitochondria was not influenced by either form of AZ. Because spermine uptake by mitochondria caused a release of cytochrome c, an enhancer of apoptosis, we looked for inhibitors of mitochondrial spermine uptake other than AZ. Cations such as Na+, K+, and Mg2+ were inhibitors of the mitochondrial uptake. It has been reported that heparan sulfate on glypican-1 plays important roles in spermine uptake by human embryonic lung fibroblasts. Heparin, but not heparan sulfate, slightly inhibited spermine uptake by FM3A cells in the absence of Mg2+ and Ca2+ but had no effect under physiological conditions in the presence of Mg2+ and Ca2+. PMID- 16263715 TI - Scanning cysteine mutagenesis analysis of Abeta-(1-40) amyloid fibrils. AB - We describe here the use of cysteine substitution mutants in the Alzheimer disease amyloid plaque peptide Abeta-(1-40) to probe amyloid fibril structure and stabilization. In one approach, amyloid fibrils were grown from Cys mutant peptides under reducing conditions and then challenged with an alkylating agent to probe solvent accessibility of different residues in the fibril. In another approach, monomeric Cys mutants, either in the thiol form or modified with iodoacetic acid or methyl iodide, were grown into amyloid fibrils, and the equilibrium position at the end of the amyloid formation reaction was quantified by determining the concentration of monomeric Abeta. The DeltaG values of fibril elongation obtained were then compared in order to provide information on the environment of each residue side chain in the fibril. In general, Cys residues in the N and C termini of Abeta-(1-40) were not only accessible to alkylation in the fibril state but also, when modified in the monomeric state, did not greatly impact fibril stability; these observations were consistent with previous indications that these portions of the peptide are not part of the amyloid core. In contrast, residues 16-19 and 31-34 were not only uniformly inaccessible to alkylation in the fibril state, but their modification with the negatively charged carboxymethyl group in monomeric Abeta also destabilized fibril elongation, confirming other data showing that these segments are likely packed into a hydrophobic amyloid core. Residues 20, 30, and 35, flanking these implicated beta-sandwich regions, are accessible to alkylation in the fibril indicating a location in solvent exposed structure. PMID- 16263716 TI - Regulation of luteinizing hormone receptor expression: evidence of translational suppression in vitro by a hormonally regulated mRNA-binding protein and its endogenous association with luteinizing hormone receptor mRNA in the ovary. AB - Our previous studies have identified a luteinizing hormone receptor (LHR) mRNA binding protein (LRBP) that binds to the coding region (LBS) of rat LHR mRNA. The identity of LRBP was later established as mevalonate kinase (MVK). The present study examined if LRBP binding to LHR mRNA impairs translation. A full-length FLAG-tagged rat LHR mRNA was synthesized and translated in vitro. The translation product was immunoprecipitated and analyzed on SDS-PAGE. The addition of LRBP inhibited LHR mRNA translation. This inhibitory effect was reversed by an excess of wild type (wt) LBS. To determine whether this reversal of the inhibitory effect of LRBP was indeed due to the sequestration of LRBP by the wtLBS, a translation reaction was performed in the presence of mutated LBS in which all the cytidine in the wtLBS was mutated to uridine. This mutation of LBS has been shown to render it incapable of interacting with LRBP. Unlike wtLBS, the mutated LBS was unable to reverse the inhibitory effect of LRBP on LHR mRNA translation. The addition of mevalonate, which has been shown to compete for LHR mRNA binding to LRBP, also reduced the extent of translation inhibition by LRBP. Endogenous association of LHR mRNA with MVK was assessed by immunoprecipitation of the ribonucleoprotein complex with MVK antibody followed by reverse transcription-PCR of the RNA associated with the immune complex. Amplification of LHR mRNA, if any, associated with the immunoprecipitate obtained from ovarian ribonucleoprotein complex with gene-specific primers confirmed the association of LHR mRNA with MVK. Collectively, the present data support the novel function of LRBP as a translational inhibitor of LHR mRNA in the ovary. PMID- 16263717 TI - c-Myb and members of the c-Ets family of transcription factors act as molecular switches to mediate opposite steroid regulation of the human glucocorticoid receptor 1A promoter. AB - Steroid auto-regulation of the human glucocorticoid receptor (hGR) 1A promoter in lymphoblast cells resides largely in two DNA elements (footprints 11 and 12). We show here that c-Myb and c-Ets family members (Ets-1/2, PU.1, and Spi-B) control hGR 1A promoter regulation in T- and B-lymphoblast cells. Two T-lymphoblast lines, CEM-C7 and Jurkat, contain high levels of c-Myb and low levels of PU.1, whereas the opposite is true in IM-9 B-lymphoblasts. In Jurkat cells, overexpression of c-Ets-1, c-Ets-2, or PU.1 effectively represses dexamethasone mediated up-regulation of an hGR 1A promoter-luciferase reporter gene, as do dominant negative c-Myb (c-Myb DNA-binding domain) or Ets proteins (Ets-2 DNA binding domain). Overexpression of c-Myb in IM-9 cells confers hormone-dependent up-regulation to the hGR 1A promoter reporter gene. Chromatin immunoprecipitation assays show that hormone treatment causes the recruitment of hGR and c-Myb to the hGR 1A promoter in CEM-C7 cells, whereas hGR and PU.1 are recruited to this promoter in IM-9 cells. These observations suggest that the specific transcription factor that binds to footprint 12, when hGR binds to the adjacent footprint 11, determines the direction of hGR 1A promoter auto-regulation. This leads to a "molecular switch" model for auto-regulation of the hGR 1A promoter. PMID- 16263719 TI - Functional human mitochondrial DNA polymerase gamma forms a heterotrimer. AB - Mitochondrial DNA polymerase gamma (pol gamma) is responsible for replication and repair of mtDNA and is mutated in individuals with genetic disorders such as chronic external ophthalmoplegia and Alpers syndrome. pol gamma is also an adventitious target for toxic side effects of several antiviral compounds, and mutation of its proofreading exonuclease leads to accelerated aging in mouse models. We have used a variety of physical and functional approaches to study the interaction of the human pol gamma catalytic subunit with both the wild-type accessory factor, pol gammaB, and a deletion derivative that is unable to dimerize and consequently is impaired in its ability to stimulate processive DNA synthesis. Our studies clearly showed that the functional human holoenzyme contains two subunits of the processivity factor and one catalytic subunit, thereby forming a heterotrimer. The structure of pol gamma seems to be variable, ranging from a single catalytic subunit in yeast to a heterodimer in Drosophila and a heterotrimer in mammals. PMID- 16263718 TI - How dihydrolipoamide dehydrogenase-binding protein binds dihydrolipoamide dehydrogenase in the human pyruvate dehydrogenase complex. AB - The dihydrolipoamide dehydrogenase-binding protein (E3BP) and the dihydrolipoamide acetyltransferase (E2) component enzyme form the structural core of the human pyruvate dehydrogenase complex by providing the binding sites for two other component proteins, dihydrolipoamide dehydrogenase (E3) and pyruvate dehydrogenase (E1), as well as pyruvate dehydrogenase kinases and phosphatases. Despite a high similarity between the primary structures of E3BP and E2, the E3 binding domain of human E3BP is highly specific to human E3, whereas the E1 binding domain of human E2 is highly specific to human E1. In this study, we characterized binding of human E3 to the E3-binding domain of E3BP by x-ray crystallography at 2.6-angstroms resolution, and we used this structural information to interpret the specificity for selective binding. Two subunits of E3 form a single recognition site for the E3-binding domain of E3BP through their hydrophobic interface. The hydrophobic residues Pro133, Pro154, and Ile157 in the E3-binding domain of E3BP insert themselves into the surface of both E3 polypeptide chains. Numerous ionic and hydrogen bonds between the residues of three interacting polypeptide chains adjacent to the central hydrophobic patch add to the stability of the subcomplex. The specificity of pairing for human E3BP with E3 is interpreted from its subcomplex structure to be most likely due to conformational rigidity of the binding fragment of the E3-binding domain of E3BP and its exquisite amino acid match with the E3 target interface. PMID- 16263720 TI - Molecular architecture and ligand recognition determinants for T4 RNA ligase. AB - RNA ligase type 1 from bacteriophage T4 (Rnl1) is involved in countering a host defense mechanism by repairing 5'-PO4 and 3'-OH groups in tRNA(Lys). Rnl1 is widely used as a reagent in molecular biology. Although many structures for DNA ligases are available, only fragments of RNA ligases such as Rnl2 are known. We report the first crystal structure of a complete RNA ligase, Rnl1, in complex with adenosine 5'-(alpha,beta-methylenetriphosphate) (AMPcPP). The N-terminal domain is related to the equivalent region of DNA ligases and Rnl2 and binds AMPcPP but with further interactions from the additional N-terminal 70 amino acids in Rnl1 (via Tyr37 and Arg54) and the C-terminal domain (Gly269 and Asp272). The active site contains two metal ions, consistent with the two magnesium ion catalytic mechanism. The C-terminal domain represents a new all alpha-helical fold and has a charge distribution and architecture for helix nucleic acid groove interaction compatible with tRNA binding. PMID- 16263721 TI - Hsk1-Dfp1/Him1, the Cdc7-Dbf4 kinase in Schizosaccharomyces pombe, associates with Swi1, a component of the replication fork protection complex. AB - The protein kinase Hsk1 is essential for DNA replication in Schizosaccharomyces pombe. It associates with Dfp1/Him1 to form an active complex equivalent to the Cdc7-Dbf4 protein kinase in Saccharomyces cerevisiae. Swi1 and Swi3 are subunits of the replication fork protection complex in S. pombe that is homologous to the Tof1-Csm3 complex in S. cerevisiae. The fork protection complex helps to preserve the integrity of stalled replication forks and is important for activation of the checkpoint protein kinase Cds1 in response to fork arrest. Here we describe physical and genetic interactions involving Swi1 and Hsk1-Dfp1/Him1. Dfp1/Him1 was identified in a yeast two-hybrid screen with Swi1. Hsk1 and Dfp1/Him1 both co immunoprecipitate with Swi1. Swi1 is required for growth of a temperature sensitive hsk1 (hsk1ts) mutant at its semi-permissive temperature. Hsk1ts cells accumulate Rad22 (Rad52 homologue) DNA repair foci at the permissive temperature, as previously observed in swi1 cells, indicating that abnormal single-stranded DNA regions form near the replication fork in hsk1ts cells. hsk1ts cells were also unable to properly delay S-phase progression in the presence of a DNA alkylating agent and were partially defective in mating type switching. These data suggest that Hsk1-Dfp1/Him1 and Swi1-Swi3 complexes have interrelated roles in stabilization of arrested replication forks. PMID- 16263722 TI - WNK1 regulates phosphorylation of cation-chloride-coupled cotransporters via the STE20-related kinases, SPAK and OSR1. AB - The WNK1 and WNK4 genes have been found to be mutated in some patients with hyperkalemia and hypertension caused by pseudohypoaldosteronism type II. The clue to the pathophysiology of pseudohypoaldosteronism type II was its striking therapeutic response to thiazide diuretics, which are known to block the sodium chloride cotransporter (NCC). Although this suggests a role for WNK1 in hypertension, the precise molecular mechanisms are largely unknown. Here we have shown that WNK1 phosphorylates and regulates the STE20-related kinases, Ste20 related proline-alanine-rich kinase (SPAK) and oxidative stress response 1 (OSR1). WNK1 was observed to phosphorylate the evolutionary conserved serine residue located outside the kinase domains of SPAK and OSR1, and mutation of the OSR1 serine residue caused enhanced OSR1 kinase activity. In addition, hypotonic stress was shown to activate SPAK and OSR1 and induce phosphorylation of the conserved OSR1 serine residue, suggesting that WNK1 may be an activator of the SPAK and OSR1 kinases. Moreover, SPAK and OSR1 were found to directly phosphorylate the N-terminal regulatory regions of cation-chloride-coupled cotransporters including NKCC1, NKCC2, and NCC. Phosphorylation of NCC was induced by hypotonic stress in cells. These results suggested that WNK1 and SPAK/OSR1 mediate the hypotonic stress signaling pathway to the transporters and may provide insights into the mechanisms by which WNK1 regulates ion balance. PMID- 16263723 TI - Targeting of unfolded PhoA to the TAT translocon of Escherichia coli. AB - In Escherichia coli, the Tat system does not translocate Tat signal sequence fused PhoA (RR-PhoA), as it requires disulfide formation for folding. Here we show that such a RR-PhoA construct can be efficiently targeted to the Tat translocon, but the transport is not completed. RR-PhoA is detectable in a 580 kDa TatBC-containing complex, which is the first substrate-bound TatBC complex detected in a bacterial system so far. A second TatBC complex near 440 kDa comprises most of the TatB and TatC but is devoid of RR-PhoA. The targeting of PhoA to the Tat translocon depends on the twin-arginine motif and results in severe growth defects. This physiological effect is likely to be due to proton leakage at the cytoplasmic membrane. The results point to mechanistic incompatibilities of the Tat system with unfolded proteins such as RR-PhoA. There does not exist an intrinsic quality control at the TatBC complex itself, although correct folding is inevitable for Tat-dependent translocation. PMID- 16263724 TI - Receptor-type protein-tyrosine phosphatase-kappa regulates epidermal growth factor receptor function. AB - Epidermal growth factor receptor (EGFR), the prototypic receptor protein tyrosine kinase, is a major regulator of growth and survival for many epithelial cell types. We report here that receptor-type protein-tyrosine phosphatase-kappa (RPTP kappa) dephosphorylates EGFR and thereby regulates its function in human keratinocytes. Protein-tyrosine phosphatase (PTP) inhibitors induced EGFR tyrosine phosphorylation in intact primary human keratinocytes and cell-free membrane preparations. Five highly expressed RPTPs (RPTP-beta, delta, kappa, mu, and xi) were functionally analyzed in a Chinese hamster ovary (CHO) cell-based expression system. Full-length human EGFR expressed in CHO cells, which lack endogenous EGFR, displayed high basal (i.e. in the absence of ligand) tyrosine phosphorylation. Co-expression of RPTP-kappa, but not other RPTPs, specifically reduced basal EGFR tyrosine phosphorylation. RPTP-kappa also reduced epidermal growth factor-dependent EGFR tyrosine phosphorylation in CHO cells. Purified RPTP kappa preferentially dephosphorylated EGFR tyrosines 1068 and 1173 in vitro. Overexpression of wild-type or catalytically inactive RPTP-kappa reduced or enhanced, respectively, basal and EGF-induced EGFR tyrosine phosphorylation in human keratinocytes. Furthermore, siRNA-mediated knockdown of RPTP-kappa increased basal and EGF-stimulated EGFR tyrosine phosphorylation and augmented downstream Erk activation in human keratinocytes. RPTP-kappa levels increased in keratinocytes as cells reached confluency, and overexpression of RPTP-kappa in subconfluent keratinocytes reduced keratinocyte proliferation. Taken together, the above data indicate that RPTP-kappa is a key regulator of EGFR tyrosine phosphorylation and function in human keratinocytes. PMID- 16263725 TI - Discovery of small molecule inhibitors of the interaction of the thyroid hormone receptor with transcriptional coregulators. AB - Thyroid hormone (3,5,3'-triiodo-L-thyronine, T3) is an endocrine hormone that exerts homeostatic regulation of basal metabolic rate, heart rate and contractility, fat deposition, and other phenomena (1, 2). T3 binds to the thyroid hormone receptors (TRs) and controls their regulation of transcription of target genes. The binding of TRs to thyroid hormone induces a conformational change in TRs that regulates the composition of the transcriptional regulatory complex. Recruitment of the correct coregulators (CoR) is important for successful gene regulation. In principle, inhibition of the TR-CoR interaction can have a direct influence on gene transcription in the presence of thyroid hormones. Herein we report a high throughput screen for small molecules capable of inhibiting TR coactivator interactions. One class of inhibitors identified in this screen was aromatic beta-aminoketones, which exhibited IC50 values of approximately 2 microm. These compounds can undergo a deamination, generating unsaturated ketones capable of reacting with nucleophilic amino acids. Several experiments confirm the hypothesis that these inhibitors are covalently bound to TR. Optimization of these compounds produced leads that inhibited the TR-CoR interaction in vitro with potency of approximately 0.6 microm and thyroid signaling in cellular systems. These are the first small molecules irreversibly inhibiting the coactivator binding of a nuclear receptor and suppressing its transcriptional activity. PMID- 16263726 TI - Human but not yeast CHD1 binds directly and selectively to histone H3 methylated at lysine 4 via its tandem chromodomains. AB - Defining the protein factors that directly recognize post-translational, covalent histone modifications is essential toward understanding the impact of these chromatin "marks" on gene regulation. In the current study, we identify human CHD1, an ATP-dependent chromatin remodeling protein, as a factor that directly and selectively recognizes histone H3 methylated on lysine 4. In vitro binding studies identified that CHD1 recognizes di- and trimethyl H3K4 with a dissociation constant (Kd) of approximately 5 microm, whereas monomethyl H3K4 binds CHD1 with a 3-fold lower affinity. Surprisingly, human CHD1 binds to methylated H3K4 in a manner that requires both of its tandem chromodomains. In vitro analyses demonstrate that unlike human CHD1, yeast Chd1 does not bind methylated H3K4. Our findings indicate that yeast and human CHD1 have diverged in their ability to discriminate covalently modified histones and link histone modification-recognition and non-covalent chromatin remodeling activities within a single human protein. PMID- 16263728 TI - Extrinsic compression of the left main coronary artery. PMID- 16263727 TI - Creation of GPCR-based chemical sensors by directed evolution in yeast. AB - G protein-coupled receptors (GPCRs) form a class of biological chemical sensors with an enormous diversity in ligand binding and sensitivity. To explore structural aspects of ligand recognition, we subjected the human UDP-glucose receptor (P2Y14) functionally expressed in the yeast Saccharomyces to directed evolution. We sought to generate new receptor subtypes with ligand-binding properties that would be useful in the development of practical biosensors. Mutagenesis of the entire UDP-glucose receptor gene yielded receptors with increased activity but similar ligand specificities, while random mutagenesis of residues in the immediate vicinity of the ligand-binding pocket yielded mutants with altered ligand specificity. By first sensitizing the P2Y14 receptor and then redirecting ligand specificity, we were able to create mutant receptors suitable for a simple biosensor. Our results demonstrate the feasibility of altering receptor ligand-binding properties via a directed evolution strategy, using standard yeast genetic techniques. The novel receptor mutants can be used to detect chemical ligands in complex mixtures and to discriminate among chemically or stereochemically related compounds. Specifically, we demonstrate how engineered receptors can be applied in a pairwise manner to differentiate among several chemical analytes that would be indistinguishable with a single receptor. These experiments demonstrate the feasibility of a combinatorial approach to detector design based on the principles of olfaction. PMID- 16263729 TI - CMR scarring in a patient with hypertrophic cardiomyopathy correlates well with histological findings of fibrosis. PMID- 16263730 TI - In vivo quantification of central venous catheter leak. PMID- 16263731 TI - Brachial artery thrombosis due to haemodialysis arteriovenous fistula. PMID- 16263732 TI - The role of gender in estimating glomerular filtration rate. PMID- 16263733 TI - Lamivudine and HBV-associated nephropathy. PMID- 16263734 TI - Hypertensive myocardial fibrosis. PMID- 16263736 TI - Sheehan syndrome presented with acute renal failure associated with rhabdomyolysis and hyponatraemia. PMID- 16263735 TI - Vascular calcification and cardiovascular function in chronic kidney disease. AB - BACKGROUND: Vascular calcification and arterial stiffening are independent predictors of all causes and cardiovascular mortality in chronic kidney disease (CKD). Few data are currently available comparing vascular calcification and its attendant functional cardiovascular consequences between CKD stage 4 patients and both peritoneal dialysis (PD) and haemodialysis (HD) (CKD stage 5) patients. METHOD: We studied 134 subjects (60 HD, 28 PD and 46 CKD 4). Vascular calcification was quantified using multi-slice spiral CT scanning of a 5 cm standardized segment of superficial femoral artery. Pulse wave analysis and pulse wave velocity were assessed using applanation tonometry, to determine arterial compliance. Further digital arterial pulse wave analysis was used to measure systemic haemodynamic variables. All medications were recorded and biochemical variables were time averaged for the 6 months prior to entering the study. RESULTS: Forty-seven percent of CKD 4 patients demonstrated vascular calcification as compared with CKD 5 (71% PD and 73% HD, P = 0.02). HD patients had higher calcification scores (median 121) than either PD (median 21) or CKD 4 (median 0) (P = 0.008). There were no significant differences in baseline characteristics between the groups. Comparing tertiles of patients (based on calcification score), increased calcification score was associated with a reduction in arterial compliance (mean PWV 8.9 +/- 1.1, 11 +/- 3.6, 11.3 +/- 3.7 m/s, P = 0.005). The degree of calcification did not influence systolic blood pressure (BP), diastolic BP or heart rate. However, more heavily calcified patients demonstrated significantly higher mean pulse pressures (58 +/- 19, 74 +/ 22 and 72 +/- 25 mmHg, P = 0.001), lower total peripheral resistance (1.5 +/- 1, 1.3 +/- 0.8, 0.9 +/- 0.4, P = 0.01) and higher stroke volume (84 +/- 25, 95 +/- 29, 106 +/- 39 ml, P = 0.01). More heavily calcified patients were significantly older and predominantly male. CONCLUSION: This study has successfully utilized a novel technique for the quantification of calcification. We have demonstrated vascular calcification and associated cardiovascular dysfunction in CKD 4, PD and HD with significant differences between the groups. Thirty percent of individuals show no calcification, even those established on renal replacement therapy for a prolonged period of time. Further work is required to identify factors which promote progression of arterial calcification in those who are susceptible. PMID- 16263737 TI - Is it practical to screen dialysis patients for vascular calcification? PMID- 16263738 TI - Hypothalamic neurocysticercosis presenting with polyuria: a first report of an unusual manifestation. PMID- 16263739 TI - Prevalence, clinical correlates and therapy cost of mineral abnormalities among haemodialysis patients: a cross-sectional multicentre study. AB - BACKGROUND: This study evaluated the proportion of patients who met National Kidney Foundation Kidney Disease Outcomes Quality Initiative (NKF-K/DOQI) guidelines for mineral status, and assessed the cost of therapy for mineral management of patients under haemodialysis treatment in Spain. METHODS: Demographic and biochemical data were collected for 1312 patients undergoing standard three-times weekly maintenance haemodialysis at six Spanish centres during December 2003. Age, gender, diabetic nephropathy, haemodialysis duration, serum calcium, phosphorus, calcium-phosphorus product (Ca x P), and intact parathyroid hormone (iPTH) levels were monitored. Exploratory analyses of associations between demographic and biochemical parameters, were undertaken using bivariate and multivariate regression techniques. RESULTS: Mean age of patients was 62 years. 97% were Caucasian, 23% were diabetic. In total, 51% of patients received calcium binders, 21% sevelamer, 16% aluminium hydroxide, and 29% received no binders; 33% of patients received calcitriol. Prevalence of patients outside K/DOQI targets was: calcium 50%, phosphorus 46%; Ca x P 33%; iPTH 77%. Elevated phosphorus (>5.5 mg/dl) was independently associated with younger age [OR 0.972 (95% CI 0.963-0.980), P<0.001] and higher iPTH [OR 1.0005 (95% CI 1.0002-1.0008), P<0.001]. Elevated Ca x P (>or=55 mg(2) x dl(2)) showed a similar relationship. High iPTH levels (>300 pmol/l) were associated with female gender [OR 1.574 (95% CI 1.213-2.041), P<0.001], high serum phosphorus [OR 1.230 (95% CI 1.130-1.338), P<0.001], and longer duration of dialysis [OR 1.003 (95% CI 1.001-1.005), P<0.01]. Poorly controlled serum phosphorus, Ca x P and iPTH were associated with more expensive therapy for mineral management. CONCLUSIONS: One in three haemodialysis patients in Spain remains above the upper target range defined in current mineral metabolism guidelines. This abnormal profile is more common in younger patients and females and therapy is more expensive in younger patients. PMID- 16263740 TI - Role of receptor for advanced glycation end-products and signalling events in advanced glycation end-product-induced monocyte chemoattractant protein-1 expression in differentiated mouse podocytes. AB - BACKGROUND: Upregulation of local monocyte chemoattractant protein-1 (MCP-1) production is involved in glomerular damage through macrophage recruitment and activation in diabetic nephropathy. Treatment of db/db mice with soluble receptor for advanced glycation end-products (RAGE) prevented recruitment of macrophages to the glomeruli and reduced albuminuria, suggesting that binding of ligands and RAGE may be involved in MCP-1 expression. Therefore, we investigated the role of advanced glycation end-products (AGEs) in MCP-1 production by podocytes and signalling events after RAGE activation. METHODS: MCP-1 gene and protein expression were examined by using reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay in differentiated mouse podocytes. Dichlorofluorescein-sensitive intracellular reactive oxygen species (ROS) generation was measured by confocal microscopy. RAGE, phosphorylation of mitogen activated protein kinases, nuclear factor (NF)-kappaB, c-Jun and Sp1 were studied using western blotting and immunocytochemistry. RESULTS: Both differentiated and undifferentiated podocytes expressed RAGE. MCP-1 was induced by AGEs and carboxymethyllysine (CML) in a time-dependent and dose-dependent manner in differentiated podocytes. Neutralizing antibody for RAGE suppressed AGE- and CML induced MCP-1 production. AGEs and CML rapidly generated intracellular ROS in podocytes. Blocking of ROS by using N-acetyl-l-cysteine abolished CML and H(2)O(2)-induced MCP-1 expression. Phosphorylated extracellular signal-regulated kinase (ERK) was found in podocytes incubated with CML and was prevented by N acetyl-l-cysteine or 7'-amino 4 [trifluoromethyl]. PD98059, an inhibitor of ERK, partially prevented CML-induced MCP-1 gene expression. NF-kappaB and Sp1 were translocated into the nucleus after podocytes were incubated with CML for 60 min. Parthenolide and mithramycin A, inhibitors of NF-kappaB and Sp1, respectively, abolished CML-induced MCP-1 gene expression in a dose-dependent manner. CONCLUSIONS: These results suggest that AGEs and CML induce MCP-1 expression in podocytes through activation of RAGE and generation of intracellular ROS. NF kappaB and Sp1 regulate MCP-1 gene transcription. PMID- 16263741 TI - Improved biocompatibility of bicarbonate/lactate-buffered PDF is not related to pH. AB - BACKGROUND: Chronic exposure to conventional peritoneal dialysis fluid (PDF) is associated with functional and structural alterations of the peritoneal membrane. The bioincompatibility of conventional PDF can be due to hypertonicity, high glucose concentration, lactate buffering system, presence of glucose degradation products (GDPs) and/or acidic pH. Although various investigators have studied the sole effects of hyperosmolarity, high glucose, GDPs and lactate buffer in experimental PD, less attention has been paid to the chronic impact of low pH in vivo. METHODS: Rats received daily 10 ml of either conventional lactate-buffered PDF (pH 5.2; n=7), a standard bicarbonate/lactate-buffered PDF with physiological pH (n=8), bicarbonate/lactate-buffered PDF with acidic pH (adjusted to pH 5.2 with 1 N hydrochloride, n=5), or bicarbonate/lactate buffer, without glucose, pH 7.4 (n=7). Fluids were instilled via peritoneal catheters connected to implanted subcutaneous mini vascular access ports for 8 weeks. Control animals with or without peritoneal catheters served as control groups (n=8/group). Various functional (2 h PET) and morphological/cellular parameters were analyzed. RESULTS: Compared with control groups and the buffer group, conventional lactate buffered PDF induced a number of morphological/cellular changes, including angiogenesis and fibrosis in various peritoneal tissues (all parameters P<0.05), accompanied by increased glucose absorption and reduced ultrafiltration capacity. Daily exposure to standard or acidified bicarbonate/lactate-buffered PDF improved the performance of the peritoneal membrane, evidenced by reduced new vessel formation in omentum (P<0.02) and parietal peritoneum (P<0.008), reduced fibrosis (P<0.02) and improved ultrafiltration capacity. No significant differences were found between standard and acidified bicarbonate/lactate-buffered PDF. During PET, acidic PDF was neutralized within 15 to 20 min. CONCLUSION: The bicarbonate/lactate-buffered PDF, acidity per se did not contribute substantially to peritoneal worsening in our in vivo model for PD, which might be explained by the buffering capacity of the peritoneum. PMID- 16263742 TI - Treatment of peritoneal dialysis related fungal peritonitis with caspofungin plus amphotericin B combination therapy. PMID- 16263744 TI - European best practice guidelines for peritoneal dialysis. 4 Continuous ambulatory peritoneal dialysis delivery systems. PMID- 16263745 TI - European best practice guidelines for peritoneal dialysis. 5 Peritoneal dialysis solutions. PMID- 16263746 TI - European best practice guidelines for peritoneal dialysis. 1 General guidelines. PMID- 16263747 TI - European best practice guidelines for peritoneal dialysis. 6 Automated peritoneal dialysis. PMID- 16263748 TI - European best practice guidelines for peritoneal dialysis. 7 Adequacy of peritoneal dialysis. PMID- 16263749 TI - European best practice guidelines for peritoneal dialysis. 8 Nutrition in peritoneal dialysis. PMID- 16263751 TI - European best practice guidelines for peritoneal dialysis. 9 PD and transplantation. PMID- 16263750 TI - European best practice guidelines for peritoneal dialysis. 2 The initiation of dialysis. PMID- 16263753 TI - European best practice guidelines for peritoneal dialysis. 3 Peritoneal access. PMID- 16263754 TI - CpG oligodeoxynucleotides induce IL-8 expression in CD34+ cells via mitogen activated protein kinase-dependent and NF-kappaB-independent pathways. AB - To elucidate the role of Toll-like receptor 9 (TLR9) activation along with the intracellular signaling pathways triggered by CpG DNA in CD34+ cells, we investigated whether synthetic oligodeoxynucleotides (ODNs), containing unmethylated CpG motifs, could induce IL-8 expression in CD34+ cells through mitogen-activated protein kinase (MAPK) or nuclear factor-kappaB (NF-kappaB) pathway. We demonstrated evidence for the first time that CD34+ cells constitutively expressed TLR9. Exposure of the cells to CpG ODN resulted in a time- and dose-dependent increase of IL-8 expression, and activation of phosphorylated ERK1/2 and phosphorylated p38. In addition, CpG ODN stimulated AP 1, but not NF-kappaB, signals. Moreover, inhibitors of MAPK (U0126 and SB203580) significantly reduced the IL-8 production, while the inhibition of NF-kappaB (pyrrolidinedithiocarbamate and retrovirus containing dominant-negative IkappaB alpha plasmid) did not affect the IL-8 expression increased by CpG ODN. Moreover, co-stimulation with LPS and CpG synergistically up-regulates IL-8 in CD34+ cells. These results suggest that CpG DNA, acting on TLR9, activates CD34+ cells to express IL-8 through MAPK-dependent and NF-kappaB-independent pathways. PMID- 16263755 TI - Loss of CD8 and TCR binding to Class I MHC ligands following T cell activation. AB - The capacity of T cells to bind peptide/MHC ligands changes with T cell development and differentiation. Here we study changes in peptide/MHC multimer binding following T cell activation. Surprisingly, T cell activation caused a marked reduction in specific peptide/MHC Class I multimer binding, which was distinct from transient TCR down-regulation, and was especially dramatic for engagement with low-affinity peptide/MHC ligands. Direct CD8-Class I interactions were also profoundly and rapidly impaired following T cell stimulation, even though surface CD8alpha and CD8beta levels were unchanged after activation, suggesting that decreased CD8 co-receptor binding contributes to this effect. Finally, we show that enzymatic desialylation restores much of the multimer binding on activated T cells, suggesting that altered glycosylation may inhibit TCR/CD8 binding to peptide/MHC ligands. These radical changes in activated T cells' ability to perceive peptide/MHC ligands may contribute to selective outgrowth of clones with high affinity for the stimulatory ligand. PMID- 16263756 TI - The structure and location of SIMP/STT3B account for its prominent imprint on the MHC I immunopeptidome. AB - Proteins show drastic discrepancies in their contribution to the collection of self-peptides that shape the repertoire of CD8 T cells (MHC I self immunopeptidome). To decipher why selected proteins are the foremost sources of MHC I-associated self-peptides, we chose to study SIMP/STT3B because this protein generates very high amounts of MHC I-associated peptides in mice and humans. We show that the endoplasmic reticulum (ER)-associated degradation pathway and MHC I processing intersect at SIMP/STT3B. Relevant key features of SIMP/STT3B are its lysine-rich region, its propensity to misfold and its location in the ER membrane in close proximity to the immunoproteasome. Moreover, we show that coupling to SIMP/STT3B can be used to foster MHC I presentation of a selected peptide, here the ovalbumin peptide SIINFEKL. These data yield novel insights into relations between the cell proteome and the MHC I immunopeptidome. They suggest that the contribution of a given protein to the MHC I immunopeptidome results from the interplay of at least three factors: the presence of degrons (degradation signals), the tendency of the protein to misfold and its subcellular localization. Furthermore, they indicate that substrates of the ER-associated degradation pathway may have a prominent imprint on the MHC I self immunopeptidome. PMID- 16263757 TI - Imatinib for secondary Ph+ acute lymphoblastic leukemia induces response in concomitant GBM. PMID- 16263758 TI - Analyzing focal adhesion structure by atomic force microscopy. AB - Atomic force microscopy (AFM) can produce high-resolution topographic images of biological samples in physiologically relevant environments and is therefore well suited for the imaging of cellular surfaces. In this work we have investigated focal adhesion complexes by combined fluorescence microscopy and AFM. To generate high-resolution AFM topographs of focal adhesions, REF52 (rat embryo fibroblast) cells expressing YFP-paxillin as a marker for focal adhesions were de-roofed and paxillin-positive focal adhesions subsequently imaged by AFM. The improved resolution of the AFM topographs complemented the optical images and offered ultrastructural insight into the architecture of focal adhesions. Focal adhesions had a corrugated dorsal surface formed by microfilament bundles spaced 127+/-50 nm (mean+/-s.d.) apart and protruding 118+/-26 nm over the substratum. Within focal adhesions microfilaments were sometimes branched and arranged in horizontal layers separated by 10 to 20 nm. From the AFM topographs focal adhesion volumes could be estimated and were found to range from 0.05 to 0.50 microm(3). Furthermore, the AFM topographs show that focal adhesion height increases towards the stress-fiber-associated end at an angle of about 3 degrees . Finally, by correlating AFM height information with fluorescence intensities of YFP-paxillin and F-actin staining, we show that the localization of paxillin is restricted to the ventral half of focal adhesions, whereas F-actin-containing microfilaments reside predominantly in the membrane-distal half. PMID- 16263760 TI - Endocytosis of megalin by visceral endoderm cells requires the Dab2 adaptor protein. AB - Rapid endocytosis of lipoprotein receptors involves NPxY signals contained in their cytoplasmic tails. Several proteins, including ARH and Dab2, can bind these sequences, but their importance for endocytosis may vary in different cell types. The lipoprotein receptor megalin is expressed in the visceral endoderm (VE), a polarized epithelium that supplies maternal nutrients to the early mammalian embryo. Dab2 is also expressed in the VE, and is required for embryo growth and gastrulation. Here, we show that ARH is absent from the VE, and Dab2 is required for uptake of megalin, its co-receptor cubilin, and a cubilin ligand, transferrin, from the brush border of the VE into intracellular vesicles. By making isoform-specific knock-in mice, we show that the p96 splice form of Dab2, which binds endocytic proteins, can fully rescue endocytosis. The more abundant p67 isoform, which lacks some endocytic protein binding sites, only partly rescues endocytosis. Endocytosis of cubilin is also impaired in VE and in mid gestation visceral yolk sac when p96 is absent. These studies suggest that Dab2 p96 mediates endocytosis of megalin in the VE. In addition, rescue of embryonic viability correlates with endocytosis, suggesting that endocytosis mediated by Dab2 is important for normal development. PMID- 16263759 TI - Mesd binds to mature LDL-receptor-related protein-6 and antagonizes ligand binding. AB - Wnt co-receptors LRP5 and LRP6 are two members of the low-density lipoprotein receptor family. Receptor-associated protein is not only a specialized chaperone but also a universal antagonist for members of the low-density lipoprotein receptor family. Here we test whether Mesd, a newly identified chaperone for members of the low-density lipoprotein receptor family, also binds to mature receptors at the cell surface and antagonizes ligand binding. We found that Mesd binds to cell surface LRP5 and LRP6, but not to other members of the low-density lipoprotein receptor family. Scatchard analysis revealed that Mesd binds cell surface LRP6 with high affinity (K(d) approximately 3.3 nM). Interestingly, the C terminal region of Mesd, which is absent in sequences from invertebrates, is necessary and sufficient for binding to mature LRP6, and is required for LRP6 folding. We also found that LRP6 is not a constitutively active endocytosis receptor and binding of the receptor-associated protein to LRP6 partially competes for Mesd binding. Finally, we demonstrated that Mesd antagonizes ligand binding to LRP6 at the cell surface. Together our results show that in addition to serving as a folding chaperone, Mesd can function as a receptor antagonist by inhibiting ligand binding to mature LRP6. PMID- 16263761 TI - Dishevelled (Dvl-2) activates canonical Wnt signalling in the absence of cytoplasmic puncta. AB - Dishevelled family proteins are multidomain intracellular transducers of Wnt signals. Ectopically expressed mammalian Dishevelled 2 (Dvl-2) activates downstream signalling and localises to cytoplasmic puncta. It has been suggested that these Dvl-2-containing structures correspond to intracellular vesicles and may be involved in the Wnt signal transduction process. We report that cytoplasmic puncta are primarily formed in cells expressing Dvl-2 at high levels. Lower levels of expression can activate signalling without forming puncta. The structures do not localise with markers of the early or late endocytic pathway and time-lapse analysis demonstrates that Dvl-2 puncta move in a random fashion over short distances but do not originate from the plasma membrane. Based on our findings, we propose that Dvl-2 puncta are protein aggregates that are not required for signalling. PMID- 16263762 TI - The Wnt signalling effector Dishevelled forms dynamic protein assemblies rather than stable associations with cytoplasmic vesicles. AB - Dishevelled is a crucial effector upstream in the Wnt signalling pathway, but the molecular mechanism by which it transduces the Wnt signal remains elusive. Dishevelled is a cytoplasmic protein with a strong tendency to form puncta, which correlates with its potent activity in stimulating Wnt signal transduction when overexpressed. These puncta are thought to reflect cytoplasmic vesicles. However, we show here that the mammalian Dishevelled protein Dvl2 does not colocalise with known vesicle markers for clathrin-mediated or clathrin-independent endocytic pathways. Furthermore, Dvl2 puncta do not stain with lipid dyes, indicating that these puncta do not contain membranes. Instead, our evidence from live imaging by TIRF microscopy of Dvl2 tagged with green fluorescent protein (GFP-Dvl2) revealed that these puncta move in and out of the evanescent field near the plasma membrane in an undirected fashion, and that they can grow by collision and fusion. Furthermore, high-resolution confocal microscopy and photobleaching experiments indicate that the GFP-Dvl2 puncta are protein assemblies; there is a constant exchange of GFP-Dvl2 between puncta and a diffuse cytoplasmic pool, which, therefore, are in a dynamic equilibrium with each other. The same is true for the DIX domain of Dvl2 itself and also for Axin-GFP, which equilibrates between the punctate and cytosolic pools. Our evidence indicates that Dvl2 and Axin puncta are dynamic protein assemblies rather than cytoplasmic vesicles. PMID- 16263763 TI - MIDAS/GPP34, a nuclear gene product, regulates total mitochondrial mass in response to mitochondrial dysfunction. AB - To investigate the regulatory system in mitochondrial biogenesis involving crosstalk between the mitochondria and nucleus, we found a factor named MIDAS (mitochondrial DNA absence sensitive factor) whose expression was enhanced by the absence of mitochondrial DNA (mtDNA). In patients with mitochondrial diseases, MIDAS expression was increased only in dysfunctional muscle fibers. A majority of MIDAS localized to mitochondria with a small fraction in the Golgi apparatus in HeLa cells. To investigate the function of MIDAS, we stably transfected HeLa cells with an expression vector carrying MIDAS cDNA or siRNA. Cells expressing the MIDAS protein and the siRNA constitutively showed an increase and decrease in the total mass of mitochondria, respectively, accompanying the regulation of a mitochondria-specific phospholipid, cardiolipin. In contrast, amounts of the mitochondrial DNA, RNA and proteins did not depend upon MIDAS. Thus, MIDAS is involved in the regulation of mitochondrial lipids, leading to increases of total mitochondrial mass in response to mitochondrial dysfunction. PMID- 16263764 TI - Oocytes prevent cumulus cell apoptosis by maintaining a morphogenic paracrine gradient of bone morphogenetic proteins. AB - Paracrine factors secreted by the oocyte regulate a broad range of cumulus cell functions. Characteristically, cumulus cells have a low incidence of apoptosis and we proposed that this is due to oocyte-secreted factors acting in an anti apoptotic manner. Bovine cumulus-oocyte complexes (COC) were aspirated from abattoir-derived ovaries and oocytectomized (OOX) by microsurgical removal of the oocyte. OOX were treated with doses of either denuded oocytes (DO) or various growth factors for 24 hours (+/- rFSH; 0.1 IU/ml). Proportions of apoptotic cumulus cells were assessed using TUNEL and laser confocal scanning microscopy followed by image analysis. Quantification of Bcl-2 and Bax proteins in OOX was undertaken by western analysis. Oocyte removal led to a significant increase in cumulus cell apoptosis compared with COC controls (35% versus 9% TUNEL positive, respectively; P<0.001). Levels of OOX apoptosis were significantly reversed (P<0.001) in a dose-dependent manner when co-cultured with oocytes. Furthermore, the anti-apoptotic effect of oocyte-secreted factors followed a gradient from the site of the oocyte(s). Growth differentiation factor 9 (GDF9) had no significant effect on cumulus cell apoptosis. By contrast, cumulus cell apoptosis was significantly (P<0.001) reduced by bone morphogenetic proteins (BMP) 15, 6 or 7. Accordingly, levels of anti-apoptotic Bcl-2 were high in OOX+DO and OOX+BMP15 and low with OOX+GDF9 or OOX alone, whereas the reverse was observed for pro apoptotic Bax. DO, BMP15 and BMP6 were also able to protect cumulus cells from undergoing apoptosis induced by staurosporine. FSH partially prevented apoptosis in all treatment groups (P<0.001). Follistatin and a BMP6 neutralizing antibody, which antagonized the anti-apoptotic effects of BMP15 and BMP6, respectively, whether alone or combined, blocked approximately 50% of the anti-apoptotic actions of oocytes. These results are the first to demonstrate that oocyte secreted factors, and particularly BMP15 and BMP6, maintain the low incidence of cumulus cell apoptosis by establishing a localized gradient of bone morphogenetic proteins. PMID- 16263765 TI - Downregulation of the HERG (KCNH2) K(+) channel by ceramide: evidence for ubiquitin-mediated lysosomal degradation. AB - The HERG (KCNH2) potassium channel underlies the rapid component of the delayed rectifier current (I(kr)), a current contributing to the repolarisation of the cardiac action potential. Mutations in HERG can cause the hereditary forms of the short-QT and long-QT syndromes, predisposing to ventricular arrhythmias and sudden cardiac death. HERG is expressed mainly in the cell membrane of cardiac myocytes, but has also been identified in cell membranes of a range of other cells, including smooth muscle and neurones. The mechanisms regulating the surface expression have however not yet been elucidated. Here we show, using stable HERG-expressing HEK 293 cells, that ceramide evokes a time-dependent decrease in HERG current which was not attributable to a change in gating properties of the channel. Surface expression of the HERG channel protein was reduced by ceramide as shown by biotinylation of surface proteins, western blotting and immunocytochemistry. The rapid decline in HERG protein after ceramide stimulation was due to protein ubiquitylation and its association with lysosomes. The results demonstrate that the surface expression of HERG is strictly regulated, and that ceramide modifies HERG currents and targets the protein for lysosomal degradation. PMID- 16263766 TI - A requirement for membrane cholesterol in the beta-arrestin- and clathrin dependent endocytosis of LPA1 lysophosphatidic acid receptors. AB - Lysophosphatidic acid (LPA) stimulates heterotrimeric G protein signaling by activating three closely related receptors, termed LPA(1), LPA(2) and LPA(3). Here we show that in addition to promoting LPA(1) signaling, membrane cholesterol is essential for the association of LPA(1) with beta-arrestin, which leads to signal attenuation and clathrin-dependent endocytosis of LPA(1). Reduction of clathrin heavy chain expression, using small interfering RNAs, inhibited LPA(1) endocytosis. LPA(1) endocytosis was also inhibited in beta-arrestin 1 and 2-null mouse embryo fibroblasts (beta-arrestin 1/2 KO MEFs), but was restored upon re expression of wild-type beta-arrestin 2. beta-arrestin attenuates LPA signaling as LPA(1)-dependent phosphoinositide hydrolysis was significantly elevated in beta-arrestin 1/2 KO MEFs and was reduced to wild-type levels upon re-expression of wild-type beta-arrestin. Interestingly, extraction of membrane cholesterol with methyl-beta-cyclodextrin inhibited LPA(1) signaling, beta-arrestin membrane recruitment and LPA(1) endocytosis. Cholesterol repletion restored all of these functions. However, neither the stimulation of phosphoinositide hydrolysis by the M(1) acetylcholine receptor nor its endocytosis was affected by cholesterol extraction. LPA treatment increased the detergent resistance of LPA(1) and this was inhibited by cholesterol extraction, suggesting that LPA(1) localizes to detergent-resistant membranes upon ligand stimulation. These data indicate that although LPA(1) is internalized by clathrin- and beta-arrestin dependent endocytosis, membrane cholesterol is critical for LPA(1) signaling, membrane recruitment of beta-arrestins and LPA(1) endocytosis. PMID- 16263767 TI - Glucose represses connexin36 in insulin-secreting cells. AB - The gap-junction protein connexin36 (Cx36) contributes to control the functions of insulin-producing cells. In this study, we investigated whether the expression of Cx36 is regulated by glucose in insulin-producing cells. Glucose caused a significant reduction of Cx36 in insulin-secreting cell lines and freshly isolated pancreatic rat islets. This decrease appeared at the mRNA and the protein levels in a dose- and time-dependent manner. 2-Deoxyglucose partially reproduced the effect of glucose, whereas glucosamine, 3-O-methyl-D-glucose and leucine were ineffective. Moreover, KCl-induced depolarization of beta-cells had no effect on Cx36 expression, indicating that glucose metabolism and ATP production are not mandatory for glucose-induced Cx36 downregulation. Forskolin mimicked the repression of Cx36 by glucose. Glucose or forskolin effects on Cx36 expression were not suppressed by the L-type Ca(2+)-channel blocker nifedipine but were fully blunted by the cAMP-dependent protein kinase (PKA) inhibitor H89. A 4 kb fragment of the human Cx36 promoter was identified and sequenced. Reporter gene activity driven by various Cx36 promoter fragments indicated that Cx36 repression requires the presence of a highly conserved cAMP responsive element (CRE). Electrophoretic-mobility-shift assays revealed that, in the presence of a high glucose concentration, the binding activity of the repressor CRE-modulator 1 (CREM-1) is enhanced. Taken together, these data provide evidence that glucose represses the expression of Cx36 through the cAMP-PKA pathway, which activates a member of the CRE binding protein family. PMID- 16263768 TI - Effect of exercise intensity and hypoxia on skeletal muscle AMPK signaling and substrate metabolism in humans. AB - We compared in human skeletal muscle the effect of absolute vs. relative exercise intensity on AMP-activated protein kinase (AMPK) signaling and substrate metabolism under normoxic and hypoxic conditions. Eight untrained males cycled for 30 min under hypoxic conditions (11.5% O(2), 111 +/- 12 W, 72 +/- 3% hypoxia Vo(2 peak); 72% Hypoxia) or under normoxic conditions (20.9% O(2)) matched to the same absolute (111 +/- 12 W, 51 +/- 1% normoxia Vo(2 peak); 51% Normoxia) or relative (to Vo(2 peak)) intensity (171 +/- 18 W, 73 +/- 1% normoxia Vo(2 peak); 73% Normoxia). Increases (P < 0.05) in AMPK activity, AMPKalpha Thr(172) phosphorylation, ACCbeta Ser(221) phosphorylation, free AMP content, and glucose clearance were more influenced by the absolute than by the relative exercise intensity, being greatest in 73% Normoxia with no difference between 51% Normoxia and 72% Hypoxia. In contrast to this, increases in muscle glycogen use, muscle lactate content, and plasma catecholamine concentration were more influenced by the relative than by the absolute exercise intensity, being similar in 72% Hypoxia and 73% Normoxia, with both trials higher than in 51% Normoxia. In conclusion, increases in muscle AMPK signaling, free AMP content, and glucose disposal during exercise are largely determined by the absolute exercise intensity, whereas increases in plasma catecholamine levels, muscle glycogen use, and muscle lactate levels are more closely associated with the relative exercise intensity. PMID- 16263769 TI - Meal feeding enhances formation of eIF4F in skeletal muscle: role of increased eIF4E availability and eIF4G phosphorylation. AB - Feeding promotes protein accretion in skeletal muscle through a stimulation of the mRNA translation initiation phase of protein synthesis either secondarily to nutrient-induced rises in insulin or owing to direct effects of nutrients themselves. The present set of experiments establishes the effects of meal feeding on potential signal transduction pathways that may be important in accelerating mRNA translation initiation. Gastrocnemius muscle from male Sprague Dawley rats trained to consume a meal consisting of rat chow was sampled before, during, and after the meal. Meal feeding enhanced the assembly of the active eIF4G.eIF4E complex, which returned to basal levels within 3 h of removal of food. The increased assembly of the active eIF4G.eIF4E complex was associated with a marked 10-fold rise in phosphorylation of eIF4G(Ser(1108)) and a decreased assembly of inactive 4E-BP1.eIF4E complex. The reduced assembly of 4E-BP1.eIF4E complex was associated with a 75-fold increase in phosphorylation of 4E-BP1 in the gamma-form during feeding. Phosphorylation of S6K1 on Ser(789) was increased by meal feeding, although the extent of phosphorylation was greater at 0.5 h after feeding than after 1 h. Phosphorylation of mammalian target of rapamycin (mTOR) on Ser(2448) or Ser(2481), an upstream kinase responsible for phosphorylating both S6K1 and 4E-BP1, was increased at all times during meal feeding, although the extent of phosphorylation was greater at 0.5 h after feeding than after 1 h. Phosphorylation of PKB, an upstream kinase responsible for phosphorylating mTOR, was elevated only after 0.5 h of meal feeding for Thr(308), whereas phosphorylation Ser(473) was significantly elevated at only 0.5 and 1 h after initiation of feeding. We conclude from these studies that meal feeding stimulates two signal pathways in skeletal muscle that lead to elevated eIF4G.eIF4E complex assembly through increased phosphorylation of eIF4G and decreased association of 4E-BP1 with eIF4E. PMID- 16263770 TI - Anabolic signaling and protein synthesis in human skeletal muscle after dynamic shortening or lengthening exercise. AB - We hypothesized a differential activation of the anabolic signaling proteins protein kinase B (PKB) and p70 S6 kinase (p70(S6K)) and subsequent differential stimulation of human muscle protein synthesis (MPS) after dynamic shortening or lengthening exercise. Eight healthy men [25 +/- 5 yr, BMI 26 +/- 3 kg/m(-2) (means +/- SD)] were studied before and after 12 min of repeated stepping up to knee height, and down again, while carrying 25% of their body weight, i.e., shortening exercise with the "up" leg and lengthening exercise with contralateral "down" leg. Quadriceps biopsies were taken before and 3, 6, and 24 h after exercise. After exercise, over 2 h before the biopsies, the subjects ingested 500 ml of water containing 45 g of essential amino acids and 135 g of sucrose. Rates of muscle protein synthesis were determined via incorporation over time of [1 (13)C]leucine ( or =65 yr with RA for > or =6 months were enrolled in an open-label, non controlled study. All DMARDs were discontinued and tacrolimus was administered orally once daily after the evening meal for 28 weeks. Tacrolimus, initiated at 1.5 mg/day, was increased to 3 mg/day after 6 weeks if no abnormal changes developed. Existing NSAID and oral corticosteroid (< or =7.5 mg/day prednisolone equivalent) therapy could be continued during the study. Safety was evaluated as clinical symptoms, abnormal changes in laboratory values and the development of infection. Treatment response was determined using the American College of Rheumatology (ACR) criteria for improvement. Whole blood concentrations of tacrolimus 12 h after administration were measured by high-performance liquid chromatography and tandem mass spectrometry. RESULTS: Clinical adverse events developed in 25 patients (46.3%). Abnormal changes in laboratory values occurred in 25 patients (46.3%). Ten patients (18.5%) developed infection. An ACR20 response was achieved by 50.0% of efficacy-evaluable patients and ACR20 success rates (the proportion of patients achieving ACR20 response and completing the study) was 46.3%. The ACR50 response rate was 18.5% of evaluable patients. Mean blood concentration of tacrolimus was 3.3 and 5.3 ng/ml in patients receiving 1.5 and 3.0 mg daily, respectively. No relationship between its concentration and adverse reactions was observed. CONCLUSION: In elderly patients with insufficient response to DMARD therapy, tacrolimus at 1.5-3.0 mg/day is safe and well tolerated and provides clinical benefit. PMID- 16263778 TI - Patients with stable long-standing rheumatoid arthritis continue to deteriorate despite intensified treatment with traditional disease modifying anti-rheumatic drugs--results of the British Rheumatoid Outcome Study Group randomized controlled clinical trial. AB - OBJECTIVE: Patients with rheumatoid arthritis (RA) should start treatment early with the aim of suppressing the inflammatory process completely. It is not known if this strategy should, or can, be continued in later disease. METHODS: In a multicentre, randomized, observer-blinded, controlled trial, 466 patients with established RA (>5 yr), on stable therapy for at least 6 months, were randomized to adequate symptom control/shared care setting (SCSC) or aggressive treatment/hospital setting (ATH). All were reviewed annually by a rheumatologist. The primary outcome after 3 yr was the Health Assessment Questionnaire (HAQ). Others included the OMERACT core set and the Disease Activity Score (DAS) 28. RESULTS: Three hundred and ninety-nine patients completed the trial. There was a significant deterioration in HAQ in both arms. Only the physician global score differed between the arms. CONCLUSIONS: The trial showed no additional benefit of intensified treatment with traditional disease modifying anti-rheumatic drugs (DMARDs) in patients with stable, established RA. It proved hard to suppress C reactive protein levels. Patients in the SCSC arm were able to initiate treatment changes when their symptoms deteriorated without frequent hospital assessment. Pending further evidence, the model of shared care with annual hospital review is as good as 4-monthly hospital review for these patients. PMID- 16263779 TI - A double-blind trial of depot corticosteroids in Behcet's syndrome. AB - OBJECTIVES: Corticosteroids are widely used in Behcet's syndrome despite the absence of controlled studies. We assessed the effect of depot corticosteroids primarily for genital ulcers and secondarily for the other mucocutaneous manifestations of Behcet's syndrome. METHODS: We randomized 86 patients who had active disease with genital ulcers to receive either intramuscular corticosteroid injections (40 mg methylprednisolone acetate) or placebo every 3 weeks for 27 weeks. RESULTS: Seventy-six patients (88%) completed the treatment. There were no significant differences in the mean number of genital and oral ulcers, or folliculitis between groups. The mean number of erythema nodosum lesions was less in the corticosteroid group as a whole (P = 0.0046); subgroup analyses revealed that this was significant for females (P = 0.0148) but not for males (P = 0.1). CONCLUSION: Low-dose depot corticosteroids did not have any beneficial effect on genital ulcers. However, it was useful in controlling erythema nodosum lesions, especially among the females. PMID- 16263780 TI - Musculoskeletal ultrasound training in rheumatology: the Belfast experience. AB - OBJECTIVES: Despite the increasing use of musculoskeletal ultrasound (MSUS) as a clinical tool in rheumatology, there is no consensus yet regarding the standards required to achieve a basic level of competence in the use of this imaging technique. A number of sonographers worldwide are developing curricula and standardizing teaching methods in order to improve training in MSUS for rheumatologists. In the meantime, clinicians are devising informal means of training in order to acquire these new skills. Here we describe the informal team approach to MSUS training adopted by a group of rheumatologists from the Regional Rheumatology Centre in Belfast, UK. METHODS: Over a 5-yr period, eight rheumatologists from Musgrave Park Hospital in Belfast used a variety of means to learn the basic skills of MSUS. RESULTS: Seven of the team underwent a formal assessment of their competency in a practical examination devised by an experienced sonographer. All were judged to have attained a basic competency in MSU. CONCLUSIONS: This Belfast experience shows what can be achieved despite the absence of formal MSUS training. Nevertheless, the development of recognized training programmes and international standards of competency are important goals on the way to achieving more widespread acceptance of MSUS as a useful tool in everyday clinical practice. PMID- 16263781 TI - Prevalence and incidence of adults consulting for shoulder conditions in UK primary care; patterns of diagnosis and referral. AB - OBJECTIVES: To estimate the national prevalence and incidence of adults consulting for a shoulder condition and to investigate patterns of diagnosis, treatment, consultation and referral 3 yr after initial presentation. METHODS: Prevalence and incidence rates were estimated for 658469 patients aged 18 and over in the year 2000 using a primary care database, the IMS Disease Analyzer Mediplus UK. A cohort of 9215 incident cases was followed-up prospectively for 3 yr beyond the initial consultation. RESULTS: The annual prevalence and incidence of people consulting for a shoulder condition was 2.36% [95% confidence interval (CI) 2.32-2.40%] and 1.47% (95% CI 1.44-1.50%), respectively. Prevalence increased linearly with age whilst incidence peaked at around 50 yr then remained static at around 2%. Around half of the incident cases consulted once only, while 13.6% were still consulting with a shoulder problem during the third year of follow-up. During the 3 yr following initial presentation, 22.4% of patients were referred to secondary care, 30.8% were prescribed non-steroidal anti-inflammatory drugs and 10.6% were given an injection by their general practitioner (GP). GPs tended to use a limited number of generalized codes when recording a diagnosis; just five of 426 possible Read codes relating to shoulder conditions accounted for 74.6% of the diagnoses of new cases recorded by GPs. CONCLUSIONS: The prevalence of people consulting for shoulder problems in primary care is substantially lower than community-based estimates of shoulder pain. Most referrals occur within 3 months of initial presentation, but only a minority of patients are referred to orthopaedic specialists or rheumatologists. GPs may lack confidence in applying precise diagnoses to shoulder conditions. PMID- 16263782 TI - Touch-screen computer systems in the rheumatology clinic offer a reliable and user-friendly means of collecting quality-of-life and outcome data from patients with rheumatoid arthritis. AB - OBJECTIVES: To investigate the feasibility of collecting rheumatoid arthritis (RA) patient self-administered outcome data using touch-screen computers in a routine out-patient clinic. METHODS: Forty patients with RA completed the touch screen and paper Rheumatoid Arthritis Quality of Life Questionnaire (RAQol) in the clinic and rated ease of use and preference. Forty-five others completed the Stanford Health Assessment Questionnaire (HAQ) and visual analogue scales (VASs) for pain, fatigue and global arthritis activity on touch screen and paper and a joint assessment on touch screen. They rated ease of use and willingness to complete the assessment again. Joints were independently assessed, and completion times and technical problems recorded. RESULTS: No technical problems were encountered. The touch-screen RAQol took no longer to complete, was preferred by 64% (33% had no preference) and was rated significantly higher for ease of use (two-tailed P=0.003, n=40) even by computer naive patients (two-tailed P=0.031, n=24). Intraclass correlation coefficients between methods were high for RAQol (0.986) and tender joint counts (0.918), and as high for the pain, fatigue and global activity (0.855, 0.741, 0.881) as for test-retest of the paper versions (0.865, 0.746, 0.863). Ninety-eight per cent rated the touch screen very/quite easy for HAQ and VAS, and 90% for joint assessment. Ninety-six per cent stated a willingness to complete the touch-screen assessment in clinic again. CONCLUSIONS: Touch-screen questionnaires in the clinic can produce comparable results to paper, eliminate the need for data entry and afford immediate access to results. It is an acceptable, and in many cases a preferable, option to paper, regardless of age and previous experience of computers. PMID- 16263783 TI - A simple extension to the Rheumatoid Arthritis Quality of Life Questionnaire (RAQol) to explore individual patient concerns and monitor group outcome in clinical practice. AB - OBJECTIVES: To find out if the RAQol, if extended by a qualifying question on the level of concern associated with each item, can function both as a group outcome measure and as a useful tool to identify the concerns of individual patients. METHODS: Thirty-seven rheumatoid arthritis (RA) patients completed the questionnaire before and after starting a biological therapy. One hundred and forty-five others receiving routine care completed it at baseline, weeks 12 and 13 with EuroQol VAS and questions on global arthritis impact and any other concerns. Reproducibility was assessed in all 59 participants whose condition remained stable between weeks 12 and 13. RESULTS: The RAQol score was highly reproducible (intraclass correlation coefficient 0.986, n=59), reflected global RA impact (P = 0.000, n=140), negatively correlated with EuroQol VAS (Spearman coefficient=-0.639, two-tailed significance=0.000, n=142), responsive to biological therapy (two-tailed P= 0.000) and to increased global RA impact over 12 weeks (two-tailed P=0.012, n=37), and had high internal consistency (Cronbach's alpha=0.94, n=143). The number of issues of great concern and their percentage contribution to the RAQol score were related to global arthritis impact (P=0.000 for both) and reduced by a biological therapy (two-tailed P=0.000 and 0.001 respectively). The mean kappa for consistency in identifying each item as a concern was 0.801 (range 0.633-0.921). CONCLUSIONS: Use of the 'extended' RAQol in clinical practice could provide a valid and sensitive score for monitoring group outcome and a comprehensive and consistent list of an individual's main issues of concern to assist assessment of needs in routine clinical practice. PMID- 16263784 TI - Association of radiological hand osteoarthritis with bone mineral mass: a population study. AB - OBJECTIVES: A number of previous studies have reported an inverse relationship between osteoarthritis and osteoporosis. However, the association has remained controversial because osteoarthritis in hand joints seems to associate differently from osteoarthritis in weight-bearing joints with bone mineral mass. We studied osteoarthritis in distal interphalangeal (DIP) joints and osteoarthritis in the base of the thumb (CMC-1) for their cross-sectional associations with metacarpal cortical bone mineral mass, and for their prediction of calcaneal broadband ultrasound attenuation. METHODS: A population sample of 8000 Finns aged 30 yr and over was invited to a comprehensive health examination in 1978-1980; 90% complied. Hand radiographs were taken from 3568 participants to diagnose osteoarthritis in various hand joints, and to determine two indicators of cortical bone mineral mass, the combined cortical thickness (CCT) and the metacarpal index (MCI). Calcaneal broadband ultrasound attenuation was measured 20 yr later in 340 of these participants with the Sahara sonometer. RESULTS: In the cross-sectional setting, osteoarthritis in the DIP joints and osteoarthritis in the base of the thumb (CMC-1) were significantly associated with low CCT and low MCI. These associations were proportional to the radiological severity of osteoarthritis. In the follow-up setting, symmetrical DIP osteoarthritis adjusted for age, sex, body mass index, smoking, education, workload and MCI significantly predicted low values of broadband ultrasound attenuation. CONCLUSIONS: Our results indicate a direct relation of both radiological DIP osteoarthritis and CMC-1 osteoarthritis with low cortical bone mineral mass, in proportion to the severity of osteoarthritis. The presence of symmetrical DIP osteoarthritis, a possible indicator of generalized osteoarthritis, suggests an increased risk of osteoporosis over time. PMID- 16263785 TI - Increase in bone mineral density of patients with rheumatoid arthritis treated with anti-TNF-alpha antibody: a prospective open-label pilot study. AB - OBJECTIVE: To determine the changes in bone mineral density (BMD) in patients with rheumatoid arthritis (RA; without osteoporosis) treated with infliximab. METHODS: Twenty-six patients (19 women, seven men) aged 54.2 yr (range 27-75), with persistently active RA despite a high dose of non-steroidal anti inflammatory drugs and/or treatment with methotrexate or leflunomide, were studied. Mean duration of disease was 9.8 yr. Patients receiving or having received bisphosphonates or hormone replacement therapy were excluded. The patients were treated with 3.5 mg/kg infliximab at weeks 0, 2, 6 and then every 6 8 weeks. Lumbar and femoral BMD was measured by dual-energy X-ray absorptiometry at baseline and 12 months later. Serum osteocalcin and serum crosslaps were measured at baseline (week 0) and after 12 months. Twelve patients were taking calcium (1 g/day) and vitamin D (800 IU/day). Twenty patients were receiving methotrexate (mean dose 12.5 mg/day), six patients were receiving leflunomide (mean dose 20 mg/day) and nine patients were concomitantly receiving corticosteroids at a mean daily dose of 10 mg. RESULTS: After 12 months of infliximab therapy, there was a significant increase in BMD in the spine (BMD, P < 0.001; T-score, P < 0.001; Z-score, P < 0.001) and the femoral neck (BMD, P < 0.001; T-score, P < 0.001; Z-score, P < 0.01). With regard to the root mean square average, there was a significant increase in BMD at the left femoral neck (11.6% for a root mean square of 6%) but only a trend towards improvement in the spine (2.7% for a root mean square of 4%) during the study period. There was a significant increase in osteocalcin serum levels between baseline and after 12 months (P < 0.01) and a significant decrease in the marker for bone resorption (P < 0.01) but no change in serum calcium was observed. However, the changes in markers of bone metabolism and BMD were not correlated. CONCLUSION: The data support the hypothesis that anti-TNF therapy may exert beneficial effects on bone metabolism in RA patients. PMID- 16263786 TI - "Maturational" globin switching in primary primitive erythroid cells. AB - Mammals have 2 distinct erythroid lineages. The primitive erythroid lineage originates in the yolk sac and generates a cohort of large erythroblasts that terminally differentiate in the bloodstream. The definitive erythroid lineage generates smaller enucleated erythrocytes that become the predominant cell in fetal and postnatal circulation. These lineages also have distinct globin expression patterns. Our studies in primary murine primitive erythroid cells indicate that betaH1 is the predominant beta-globin transcript in the early yolk sac. Thus, unlike the human, murine beta-globin genes are not up-regulated in the order of their chromosomal arrangement. As primitive erythroblasts mature from proerythroblasts to reticulocytes, they undergo a betaH1- to epsilony-globin switch, up-regulate adult beta1- and beta2-globins, and down-regulate zeta globin. These changes in transcript levels correlate with changes in RNA polymerase II density at their promoters and transcribed regions. Furthermore, the epsilony- and betaH1-globin genes in primitive erythroblasts reside within a single large hyperacetylated domain. These data suggest that this "maturational" betaH1- to epsilony-globin switch is dynamically regulated at the transcriptional level. Globin switching during ontogeny is due not only to the sequential appearance of primitive and definitive lineages but also to changes in globin expression as primitive erythroblasts mature in the bloodstream. PMID- 16263787 TI - Autologous stem cell transplantation for autoimmunity induces immunologic self tolerance by reprogramming autoreactive T cells and restoring the CD4+CD25+ immune regulatory network. AB - Despite a rapidly accumulating clinical experience with autologous stem cell transplantation (ASCT) as a treatment for severe refractory autoimmune disease, data on the mechanisms by which ASCT induces immune tolerance are still very scarce. In this study it is shown that ASCT restores immunologic self-tolerance in juvenile idiopathic arthritis (JIA) via 2 mechanisms. First, ASCT induces a restoration of the frequency of FoxP3 expressing CD4+CD25bright regulatory T cells (Tregs) from severely reduced numbers before ASCT to normal levels after ASCT. This recovery is due to a preferential homeostatic expansion of CD4+CD25+ Tregs during the lymphopenic phase of immunereconstitution, as measured by Ki67 and CD44 expression, and to a renewed thymopoiesis of naive mRNA FoxP3 expressing CD4+CD25+ Tregs after ASCT. Second, using artificial antigen-presenting cells to specifically isolate self-reactive T cells, we demonstrate that ASCT induces autoimmune cells to deviate from a proinflammatory phenotype (mRNA interferon gamma [IFN-gamma] and T-bet high) to a tolerant phenotype (mRNA interleukin-10 [IL-10] and GATA-3 high). These data are the first to demonstrate the qualitative immunologic changes that are responsible for the induction of immune tolerance by ASCT for JIA: the restoration of the CD4+CD25+ immune regulatory network and reprogramming of autoreactive T cells. PMID- 16263788 TI - Classical Hodgkin lymphoma is characterized by high constitutive expression of activating transcription factor 3 (ATF3), which promotes viability of Hodgkin/Reed-Sternberg cells. AB - Hodgkin/Reed-Sternberg (HRS) cells of classical Hodgkin lymphoma (cHL) display unique characteristics that discriminate cHL from other B-cell lymphomas and normal B cells. Therefore, comparative gene expression profiling of Hodgkin and non-Hodgkin B cells could lead to the identification of candidate genes that are critical for the pathogenesis of cHL. We performed microarray analysis of Hodgkin and non-Hodgkin cell lines and identified activating transcription factor 3 (ATF3), a member of the cyclic AMP response element binding protein (CREB)/ATF family, as a differentially expressed candidate gene. Extensive analysis of a large panel of cell lines, primary tumor samples, and normal tissues revealed that high expression of ATF3 is found in nearly all cases of cHL and is almost exclusively restricted to it. Selective knock-down of ATF3 by RNA interference suppressed proliferation and strongly reduced viability of Hodgkin cells. Thus, overexpression of ATF3 is a molecular hallmark of cHL that contributes to the malignant growth of HRS cells. PMID- 16263790 TI - Exogenous heat shock protein 70 binds macrophage lipid raft microdomain and stimulates phagocytosis, processing, and MHC-II presentation of antigens. AB - The extracellular presence of endotoxin-free heat shock protein 70 (HSP70) enhances the rate and capacity of macrophage-mediated phagocytosis at 6 times the basal rate. It is protein-specific, dose- and time-dependent and involves the internalization of inert microspheres, Gram-positive and -negative bacteria and fungi. Structurally, exogenous HSP70 binds the macrophage plasma membrane, specifically on its lipid raft-microdomain. Disruption of lipid rafts, HSP70-LR interaction, or denaturing HSP70 abrogates the HSP-mediated increase in phagocytosis. Further, HSP70-mediated phagocytosis directly enhances the processing and presentation of internalized antigens via the endocytic MHC class II pathway to CD4+ T lymphocytes. Modulating the HSP70-LR interaction presents an opportunity to intervene at the level of host-pathogen interface: a therapeutic tool for emerging infections, especially when conventional treatment with antibiotics is ineffective (antibiotic resistance) or unavailable (rapidly spreading, endemic). These results identify a new role for HSP70, a highly conserved molecule in stimulating phagocytosis: a primordial macrophage function, thereby influencing both innate and adaptive immune responses. PMID- 16263789 TI - Gene-expression patterns predict phenotypes of immune-mediated thrombosis. AB - Antiphospholipid antibody syndrome (APS) is a complex autoimmune thrombotic disorder with defined clinical phenotypes. Although not all patients with elevated antiphospholipid antibody (aPLA) levels develop complications, the severity of these potential events mandates aggressive and extended lifelong anti thrombotic therapy. One hundred twenty-nine patients (57 patients with APS and venous thromboembolism [VTE], 32 patients with VTE without aPLA, 32 patients with aPLA only, and 8 healthy patients) were enrolled. RNA from peripheral-blood collection was used for DNA microarray analysis. Patterns of gene expression that characterize APS as well as thrombosis in the presence of aPLA were identified by hierarchical clustering and binary regression methods. Gene-expression profiles identify and predict individuals with APS from patients with VTE without aPLA. Importantly, similar methods identified expression profiles that accurately predicted those patients with aPLA at high risk for thrombotic events. All profiles were validated in independent cohorts of patients. The ability to predict APS, but more importantly, those patients at risk for venous thrombosis, represents a paradigm for a genomic approach that can be applied to other populations of patients with venous thrombosis, providing for more effective clinical management of disease, while also reflecting the possible underlying biologic processes. PMID- 16263791 TI - Neutrophil adhesion to endothelial cells induced by platelet factor 4 requires sequential activation of Ras, Syk, and JNK MAP kinases. AB - Signal transduction mechanisms associated with neutrophil activation by platelet factor 4 (PF4; CXCL4) are as yet poorly characterized. In a recent report, we showed that PF4-induced neutrophil functions (such as adhesion and secondary granule exocytosis) involve the activation of Src-kinases. By analyzing intracellular signals leading to adherence, we here demonstrate by several lines of evidence that in addition to Src-kinases, PF4 signaling involves the monomeric GTPase Ras, the tyrosine kinase Syk, and the MAP kinase JNK. Furthermore, on stimulation, GTPases Rac2 and RhoA were activated, and each was translocated to a different membrane compartment. As shown by inhibitor studies, Rac2 and JNK are located downstream of Syk and Ras. Most intriguingly, the latter 2 elements appear to control the activity of Rac2 and JNK independently of each other at different phases of the activation process. Although a first phase of Rac2 and JNK activation of up to 5 minutes is initiated by Ras, the second phase (5-30 minutes) depends predominantly on the activity of Syk. In summary, we describe that coordinated activity of Syk, Ras, and JNK mediates neutrophil adhesion to endothelial cells and that PF4 induces sequential activation of these elements. PMID- 16263792 TI - Repression of human gamma-globin gene expression by a short isoform of the NF-E4 protein is associated with loss of NF-E2 and RNA polymerase II recruitment to the promoter. AB - Binding of the stage selector protein (SSP) to the stage selector element (SSE) in the human gamma-globin promoter contributes to the preferential expression of the gamma-gene in fetal erythroid cells. The SSP contains the transcription factor CP2 and an erythroid-specific partner, NF-E4. The NF-E4 gene encodes a 22 kDa polypeptide employing a non-AUG initiation codon. Antisera specific to NF-E4 detects this species and an additional 14 kDa protein, which initiates from an internal methionine. Enforced expression of p14 NF-E4 in the K562 fetal/erythroid cell line, and in primary erythroid cord blood progenitors, results in repression of gamma-gene expression. Biochemical studies reveal that p14 NF-E4 interacts with CP2, resulting in diminished association of CP2 with the SSE in chromatin immunoprecipitation assays. p45 NF-E2 recruitment to the gamma-promoter is also lost, resulting in a reduction in RNA polymerase II and TBP binding and a fall in promoter transcriptional activity. This effect is specific, as enforced expression of a mutant form of p14 NF-E4, which fails to interact with CP2, also fails to repress gamma-gene expression in K562 cells. These findings provide one potential mechanism that could contribute to the autonomous silencing of the human gamma-genes in adult erythroid cells. PMID- 16263793 TI - Flk2+ myeloid progenitors are the main source of Langerhans cells. AB - Langerhans cells (LCs) are antigen-presenting cells (APCs) residing in the epidermis that play a major role in skin immunity. Our earlier studies showed that when skin is inflamed LCs are replaced by bone marrow-derived progenitor cells, while during steady-state conditions LCs are able to self-renew in the skin. Identification of the LC progenitors in bone marrow would represent a critical step toward identifying the factors that regulate LC generation as well as their trafficking to the skin. To determine LC lineage origin, we reconstituted lethally irradiated CD45.2 mice with rigorously purified lymphoid and myeloid progenitors from CD45.1 congenic mice. Twenty-four hours later, we exposed the mice to UV light to deplete resident LCs and induce their replacement by progenitors. Reconstitution with common myeloid progenitors (CMPs), common lymphoid progenitors (CLPs), granulocyte-macrophage progenitors (GMPs), or early thymic progenitors led to LC generation within 2 to 3 weeks. CMPs were at least 20 times more efficient at generating LCs than CLPs. LCs from both lineages were derived almost entirely from fetal liver kinase-2+ (Flk-2+) progenitors, displayed typical dendritic-cell (DC) morphology, and showed long-term persistence in the skin. These results indicate that LCs are derived mainly from myeloid progenitors and are dependent on Flt3-ligand for their development. PMID- 16263796 TI - Efficient stimulation of HIV-1-specific T cells using dendritic cells electroporated with mRNA encoding autologous HIV-1 Gag and Env proteins. AB - Infection with human immunodeficiency virus type 1 (HIV-1) is characterized by dysfunction of HIV-1-specific T cells. To control the virus, antigen-loaded dendritic cells (DCs) might be useful to boost and broaden HIV-specific T-cell responses. In the present study, monocyte-derived DCs from nontreated HIV-1 seropositive patients were electroporated with codon-optimized ("humanized") mRNA encoding consensus HxB-2 (hHXB-2) Gag protein. These DCs elicited a strong HIV-1 Gag-specific interferon-gamma (IFN-gamma) response by an HLA-A2-restricted CD8+ T cell line. Moreover, hHXB-2 gag mRNA-electroporated DCs also triggered IFN-gamma secretion by autologous peripheral blood mononuclear cells (PBMCs), CD4+ T cells, and CD8+ T cells from all patients tested. Next, a novel strategy was developed using autologous virus sequences. Significant specific IFN-gamma T-cell responses were induced in all patients tested by DCs electroporated with patients' autologous polymerase chain reaction (PCR)-amplified and in vitro-transcribed proviral and plasma viral mRNA encoding either Gag or Env. The stimulatory effect was seen on PBMCs, CD8+ T cells, and CD4+ T cells, demonstrating both major histocompatibility complex (MHC) class I and MHC class II antigen presentation. Moreover, a significant interleukin-2 (IL-2) T-cell response was induced by DCs electroporated with hHxB-2 or proviral gag mRNA. These findings open a major perspective for the development of patient-specific immunotherapy for HIV-1 disease. PMID- 16263794 TI - PDZ binding motif of HTLV-1 Tax promotes virus-mediated T-cell proliferation in vitro and persistence in vivo. AB - HTLV-1 cellular transformation and disease induction is dependent on expression of the viral Tax oncoprotein. PDZ is a modular protein interaction domain used in organizing signaling complexes in eukaryotic cells through recognition of a specific binding motif in partner proteins. Tax-1, but not Tax-2, contains a PDZ binding domain motif (PBM) that promotes the interaction with several cellular PDZ proteins. Herein, we investigate the contribution of the Tax-1 PBM in HTLV induced proliferation and immortalization of primary T cells in vitro and viral survival in an infectious rabbit animal model. We generated several HTLV-1 and HTLV-2 Tax viral mutants, including HTLV-1deltaPBM, HTLV-2+C22(+PBM), and HTLV-2+ C18(deltaPBM). All Tax mutants maintained the ability to significantly activate the CREB/ATF or NFkappaB signaling pathways. Microtiter proliferation assays revealed that the Tax-1 PBM significantly increases both HTLV-1- and HTLV-2 induced primary T-cell proliferation. In addition, Tax-1 PBM was responsible for the micronuclei induction activity of Tax-1 relative to that of Tax-2. Viral infection and persistence were severely attenuated in rabbits inoculated with HTLV-1deltaPBM. Our results provide the first direct evidence suggesting that PBM mediated associations between Tax-1 and cellular proteins play a key role in HTLV induced cell proliferation and genetic instability in vitro and facilitate viral persistence in vivo. PMID- 16263797 TI - Enhancement of Ca2+-regulated exocytosis by indomethacin in guinea-pig antral mucous cells: arachidonic acid accumulation. AB - Ca2+-regulated exocytosis is enhanced by an autocrine mechanism via the PGE2-cAMP pathway in antral mucous cells of guinea-pigs. The inhibition of the PGE2-cAMP pathway by H-89 (an inhibitor of protein kinase A, PKA) or aspirin (ASA, an inhibitor of cyclo-oxygenase, COX) decreased the frequency of ACh-stimulated exocytotic events by 60%. Indomethacin (IDM, an inhibitor of COX), however, decreased the frequency of ACh-stimulated exocytotic events only by 30%. Moreover, IDM increased the frequency of ACh-stimulated exocytotic events by 50% in H-89-treated or ASA-treated cells. IDM inhibits the synthesis of Prostaglandin (PGG/H) and (15R)-15-hydroxy-5,8,11 cis-13-trans-eicosatetraenoic acid (15R HPETE), while ASA inhibits only the synthesis of PGG/H. Thus, IDM may accumulate arachidonic acid (AA). AACOCF3 or N-(p-amylcinnamoyl) anthranilic acid (ACA; both inhibitors of phospholipase A2, PLA2), which inhibits AA synthesis, decreased the frequency of ACh-stimulated exocytotic events by 60%. IDM, however, did not increase the frequency in AACOCF3-treated cells. AA increased the frequency of ACh-stimulated exocytotic events in AACOCF3- or ASA-treated cells, similar to IDM in ASA- and H-89-treated cells. Moreover, in the presence of AA, IDM did not increase the frequency of ACh-stimulated exocytotic events in ASA-treated cells. The PGE2 release from antral mucosa indicates that inhibition of PLA2 by ACA inhibits the AA accumulation in unstimulated and ACh-stimulated antral mucosa. The dose-response study of AA and IDM demonstrated that the concentration of intracellular AA accumulated by IDM is less than 100 nm. In conclusion, IDM modulates the ACh-stimulated exocytosis via AA accumulation in antral mucous cells. PMID- 16263795 TI - Rho GEF Lsc is required for normal polarization, migration, and adhesion of formyl-peptide-stimulated neutrophils. AB - Neutrophil migration requires continuous reorganization of the cytoskeleton and cellular adhesion apparatus. Chemoattractants initiate intracellular signals that direct this reorganization. The signaling pathways that link chemoattractant receptors to the cytoskeleton and cellular adhesion apparatus are now being defined. Formyl-peptide chemoattractants released from bacteria stimulate G protein-linked receptors on the surface of neutrophils and regulate the neutrophil cytoskeleton and adhesion apparatus through RhoA-dependent pathways. Lsc is a RhoA guanine nucleotide exchange factor that binds the heterotrimeric G protein alpha-subunits, Galpha12 and Galpha13. We have disrupted the Lsc gene and demonstrated that formyl-peptide-stimulated Lsc knock-out (KO) neutrophils are unable to generate and sustain a single-dominant pseudopod and migrate with increased speed and reduced directionality. Unexpectedly, we also found that Lsc is required for normal beta2- and beta1-integrin-dependent neutrophil adhesion. Lsc-deficient mice have a peripheral leukocytosis and extramedullary hematopoiesis, demonstrating that Lsc is required for leukocyte homeostasis. Lsc deficient neutrophils are recruited normally to sites of bacterial peritonitis and chemical dermatitis, indicating that other signaling pathways compensate for the Lsc deficiency in some forms of inflammation. These results demonstrate that Lsc links formyl-peptide receptors to RhoA signaling pathways that regulate polarization, migration, and adhesion in neutrophils and that Lsc is required for leukocyte homeostasis. PMID- 16263798 TI - Effects of enhanced human chemosensitivity on ventilatory responses to exercise. AB - It is not clear what the effects of different types of intermittent hypoxia have on human exercise ventilation. The purpose of this study was to determine whether short-duration intermittent hypoxia, and the subsequent augmentation of the hypoxic ventilatory response (HVR), would lead to an increase in ventilatory responses during exercise at sea level. It was hypothesized that subjects exposed to short-duration intermittent hypoxia would have a greater increase in the ventilatory response to exercise compared to those exposed to long-duration intermittent hypoxia. Subjects (n = 17, male) were randomly assigned to short duration intermittent hypoxia (SDIH: 5 min of 12% O2 separated by 5 min of normoxia for 1 h) or long-duration intermittent hypoxia (LDIH: 30 min of 12% O2). Both groups had 10 exposures over a 12 day period. The HVR was measured on days 1 and 12. Maximal oxygen consumption (VO2max) was determined using a ramped cycle exercise test. Maximal exercise data were not different (P > 0.05) between SDIH and LDIH groups or following intermittent hypoxia. Minute ventilation, tidal volume and respiratory frequency were compared at 20, 40, 60, 80 and 100% of VO2max . There was no difference in the ventilatory responses at any intensity of exercise following the intermittent hypoxia period. The HVR was significantly increased following the intermittent hypoxia intervention (P < 0.05) but was not different between SDIH and LDIH (P > 0.05). The relationships between HVR and VO2max were non-significant on day 1 (r = 0.30) and day 12 (r = 0.47; P > 0.05). Our findings point to a lack of functional significance of increasing HVR via intermittent hypoxia on ventilatory responses to exercise at sea level. PMID- 16263799 TI - The interaction of central command and the exercise pressor reflex in mediating baroreflex resetting during exercise in humans. AB - Central command and the exercise pressor reflex can independently reset the carotid baroreflex (CBR) during exercise. The present investigation assessed the interactive relationship between these two neural mechanisms in mediating baroreflex resetting during exercise. Six men performed static leg exercise at 20% maximal voluntary contraction under four conditions: control, no perturbation; neuromuscular blockade (NMB) induced by administration of the neuromuscular blocking agent Norcuron (central command activation); MAST, application of medical antishock trousers inflated to 100 mmHg (exercise pressor reflex activation); and Combo, NMB plus MAST (concomitant central command and exercise pressor reflex activation). With regard to CBR control of heart rate (HR), both NMB and Combo conditions resulted in a further resetting of the carotid-cardiac stimulus-response curve compared to control conditions, suggesting that CBR-HR resetting is predominately mediated by central command. In contrast, it appears that CBR control of blood pressure can be mediated by signals from either central command or the exercise pressor reflex, since both NMB and MAST conditions equally augmented the resetting of the carotid-vasomotor stimulus-response curve. With regard to the regulation of both HR and blood pressure, the extent of CBR resetting was greater during the Combo condition than during overactivation of either central command or the exercise pressor reflex alone. Therefore, we suggest that central command and the exercise pressor reflex interact such that signals from one input facilitate signals from the other, resulting in an enhanced resetting of the baroreflex during exercise. PMID- 16263800 TI - Relationship between dietary-induced changes in intestinal commensal microflora and duodenojejunal myoelectric activity monitored by radiotelemetry in the rat in vivo. AB - Interdigestive intestinal motility, and especially phase III of the migrating myoelectric/motor complex (MMC), is responsible for intestinal clearance and plays an important role in prevention of bacterial overgrowth and translocation in the gut. Yet previous results from gnotobiotic rats have shown that intestinal microflora can themselves affect the characteristics of the myoelectric activity of the gut during the interdigestive state. Given that the composition of the intestinal microflora can be altered by dietary manipulations, we investigated the effect of supplementation of the diet with synbiotics on intestinal microflora structure and the duodenojejunal myoelectric activity in the rat. To reduce animal distress caused by restraint and handling, which can itself affect GI motility, we applied radiotelemetry for duodenojejunal EMG recordings in conscious, freely moving rats. Thirty 16-month-old Spraque-Dawley rats were used. The diet for 15 rats (E group) was supplemented with chicory inulin, Lactobacillus rhamnosus and Bifidobacterium lactis. The remaining 15 rats were fed control diet without supplements (C group). Three rats from each group were implanted with three bipolar electrodes positioned at 2, 14 and 28 cm distal to the pylorus. After recovery, two 6 h recordings of duodenojejunal EMG were carried out on each operated rat. Subsequently, group C rats received feed supplements and group E rats received only control diet for 1 week, and an additional two 6 h recordings were carried out on each of these rats. Non operated C and E rats were killed and samples of GI tract were collected for microbiological analyses. Supplementation of the diet with the pro- and prebiotics mixture increased the number of bifidobacteria, whereas it decreased the number of enterobacteria in jejunum, ileum, caecum and colon. In both caecum and colon, the dietary supplementation increased the number of total anaerobes and lactobacilli. Treatment with synbiotics increased occurrence of phase III of the MMC at all three levels of the small intestine. The propagation velocity of phase III in the whole recording segment was also increased from 3.7 +/- 0.2 to 4.4 +/- 0.2 cm min(-1) by dietary treatment. Treatment with synbiotics increased the frequency of response potentials of the propagated phase III of the MMC at both levels of the jejunum, but not in the duodenum. In both parts of the jejunum, the supplementation of the diet significantly decreased the duration of phase II of the MMC, while it did not change the duration of phase I and phase III. Using the telemetry technique it was demonstrated that changes in the gastrointestinal microflora exhibited an intestinal motility response and, more importantly, that such changes can be initiated by the addition of synbiotics to the diet. PMID- 16263801 TI - Dynamic expression of the angiotensin II type 2 receptor and duodenal mucosal alkaline secretion in the Sprague-Dawley rat. AB - Activation of angiotensin II type 2 receptors (AT2R) has been shown to stimulate duodenal mucosal alkaline secretion (DMAS) in Sprague-Dawley rats (S-D). This finding could not be confirmed in another line of S-D, and the present study investigates whether the level of AT2R expression determines the response to the AT2R agonist CGP42112A. DMAS was measured in anaesthetized rats using in situ pH stat titration. Real-time PCR and Western blot were used to assess AT1R and AT2R RNA and protein expression, respectively. CGP42112A (0.1 microg kg(-1)min(-1) I.V.) elicited a 45% net increase in DMAS in the previous S-D line studied, whereas no change occurred in the new S-D line. Luminal administration of prostaglandin E2 (10(-5) M) increased DMAS similarly in both S-D lines. AT2R protein expression was significantly higher in tissue from the previous line compared to the new line. Individual AT1R to AT2R ratios (RNA and protein) were significantly higher in the new line compared to the previous S-D line. In the new S-D line intravenous infusion of angiotensin II (Ang II; 10 microg kg(-1) h( 1)) over 120 min significantly lowered the duodenal AT1aR to AT2R RNA ratio. Prolonged Ang II infusion over 240 min increased AT2R protein expression and evoked a 42% stimulatory response in DMAS to CGP42112A. The level of local AT2R expression determines the effect of the AT2R agonist CGP42112A on rat duodenal mucosal alkaline secretion. AT2R expression should be confirmed before interpreting the experimental effects of pharmacological interferences with this receptor. PMID- 16263803 TI - Nitric oxide synthase inhibition activates L- and T-type Ca2+ channels in afferent and efferent arterioles. AB - Previous studies have shown that L-type Ca(2+) channel (LCC) blockers primarily dilate resting and ANG II-constricted afferent arterioles (AA), but do not influence either resting or ANG II-constricted efferent arterioles (EA). In contrast, blockade of T-type Ca(2+) channels (TCC) dilate EA and prevent ANG II mediated efferent constriction. The present study determined the role of LCC and TCC in mediating the AA and EA constriction following inhibition of nitric oxide synthase (NOS) and tested the hypothesis that inhibition of NOS increases the influence of LCC on EA. With the use of an isolated blood-perfused rat juxtamedullary nephron preparation, single AA or EA were visualized and superfused with a NOS inhibitor, N-nitro-l-arginine (l-NNA), with or without concomitant treatment with an LCC blocker, diltiazem, or a TCC blocker, pimozide. In response to l-NNA (1, 10, and 100 micromol/l), AA and EA diameters decreased significantly by 6.0 +/- 0.3, 13.7 +/- 1.7, and 19.9 +/- 1.4%, and by 6.2 +/- 0.5, 13.3 +/- 1.1, and 19.0 +/- 1.9%, respectively. During TCC blockade with pimozide (10 micromol/l), l-NNA did not significantly constrict afferent (0.9 +/- 0.6, 1.5 +/- 0.5, and 1.7 +/- 0.5%) or efferent (0.4 +/- 0.1, 2.1 +/- 0.7, and 2.5 +/- 1.0%) arterioles. In contrast to the responses with other vasoconstictors, the l-NNA-induced constriction of EA, as well as AA, was reversed by diltiazem (10 micromol/l). The effects were overlapping as pimozide superimposed on diltiazem did not elicit further dilation. When the effects of l NNA were reversed by superfusion with an NO donor, SNAP (10 micromol/l), diltiazem did not cause significant efferent dilation. As a further test of LCC activity, 55 mmol/l KCl, which depolarizes and constricts AA, caused only a modest constriction in resting EA (8.7 +/- 1.3%), but a stronger EA constriction during concurrent treatment with l-NNA (23.8 +/- 4.8%). In contrast, norepinephrine caused similar constrictions in both l-NNA-treated and nontreated arterioles. These results provide evidence that NO inhibits LCC and TCC activity and that NOS inhibition-mediated arteriolar constriction involves activation of LCC and TCC in both AA and EA. The difference in responses to high KCl between resting and l-NNA-constricted EA and the ability of diltiazem to block EA constriction caused by l-NNA contrasts with the lack of efferent effects in resting and SNAP-treated l-NNA-preconstricted arterioles and during ANG II mediated vasoconstriction, suggesting a recruitment of LCC in EA when NOS is inhibited. These data help explain how endothelial dysfunction associated with hypertension may lead to enhanced activity of LCC in postglomerular arterioles and increased postglomerular resistance. PMID- 16263802 TI - Glucocorticoid regulation of genes in the amiloride-sensitive sodium transport pathway by semicircular canal duct epithelium of neonatal rat. AB - The lumen of the inner ear has an unusually low concentration of endolymphatic Na+, which is important for transduction processes. We have recently shown that glucocorticoid receptors (GR) stimulate absorption of Na+ by semicircular canal duct (SCCD) epithelia. In the present study, we sought to determine the presence of genes involved in the control of the amiloride-sensitive Na+ transport pathway in rat SCCD epithelia and whether their level of expression was regulated by glucocorticoids using quantitative real-time RT-PCR. Transcripts were present for alpha-, beta-, and gamma-subunits of the epithelial sodium channel (ENaC); the alpha1-, alpha3-, beta1-, and beta3-isoforms of Na+-K+-ATPase; inwardly rectifying potassium channels [IC50 of short circuit current (Isc) for Ba2+: 210 microM] Kir2.1, Kir2.2, Kir2.3, Kir2.4, Kir3.1, Kir3.3, Kir4.1, Kir4.2, Kir5.1, and Kir7.1; sulfonyl urea receptor 1 (SUR1); GR; mineralocorticoid receptor (MR); 11beta-hydroxysteroid dehydrogenase (11beta-HSD) types 1 and 2; serum- and glucocorticoid-regulated kinase 1 (Sgk1); and neural precursor cell-expressed developmentally downregulated 4-2 (Nedd4-2). On the other hand, transcripts for the alpha4-subunit of Na+-K+-ATPase, Kir1.1, Kir3.2, Kir3.4, Kir6.1, Kir6.2, and SUR2 were found to be absent, and Isc was not inhibited by glibenclamide. Dexamethasone (100 nM for 24 h) not only upregulated the transcript expression of alpha-ENaC (approximately 4-fold), beta2-subunit (approximately 2-fold) and beta3 subunit (approximately 8-fold) of Na+-K+-ATPase, Kir2.1 (approximately 5-fold), Kir2.2 (approximately 9-fold), Kir2.4 (approximately 3-fold), Kir3.1 (approximately 3- fold), Kir3.3 (approximately 2-fold), Kir4.2 (approximately 3 fold), Kir7.1 (approximately 2-fold), Sgk1 (approximately 4-fold), and Nedd4-2 (approximately 2-fold) but also downregulated GR (approximately 3-fold) and 11beta-HSD1 (approximately 2-fold). Expression of GR and 11beta-HSD1 was higher than MR and 11beta-HSD2 in the absence of dexamethasone. Dexamethasone altered transcript expression levels (alpha-ENaC and Sgk1) by activation of GR but not MR. Proteins were present for the alpha-, beta-, and gamma-subunits of ENaC and Sgk1, and expression of alpha- and gamma-ENaC was upregulated by dexamethasone. These findings are consistent with the genomic stimulation by glucocorticoids of Na+ absorption by SCCD and provide an understanding of the therapeutic action of glucocorticoids in the treatment of Meniere's disease. PMID- 16263804 TI - Reduced autoregulatory effectiveness in adenosine 1 receptor-deficient mice. AB - Adjustments of renal vascular resistance in response to changes in blood pressure are mediated by an interplay between the myocyte-inherent myogenic and the kidney specific tubuloglomerular feedback (TGF) mechanisms. Using mice with deletion of the A(1) adenosine receptor (A1AR) gene, we tested the prediction that the absence of TGF, previously established to result from A1AR deficiency, is associated with a reduction in the efficiency of autoregulation. In anesthetized wild-type (A1AR+/+) and A1AR-deficient mice (A1AR-/-), glomerular filtration rate (GFR) and renal blood flow (RBF) were determined before and after reducing renal perfusion pressure through a suprarenal aortic clamp. In response to a blood pressure reduction by 15.9 +/- 1.34 mmHg in A1AR-/- (n = 9) and by 14.2 +/- 0.9 mmHg in A1AR+/+ mice (n = 8; P = 0.31), GFR fell by 187.9 +/- 37 mul/min and by 72.3 +/- 10 mul/min in A1AR-/- and A1AR+/+ mice, respectively (P = 0.013). Similarly, with pressure reductions of 14.8 +/- 1.1 and 13.3 +/- 1.5 mmHg in A1AR /- (n = 9) and wild-type mice (n = 8), respectively (P = 0.43), RBF fell by 0.17 +/- 0.02 ml/min in A1AR-/- mice and by only 0.08 +/- 0.02 ml/min in wild-type animals (P = 0.0039). Autoregulatory indexes for both GFR and RBF were significantly higher in A1AR-/- compared with A1AR+/+ mice, indicating reduced regulatory responsiveness in the knockout animals. We conclude that autoregulation of renal vascular resistance is less complete in A1AR-deficient mice, an effect that is presumably related to absence of TGF regulation in these animals. PMID- 16263805 TI - Immunolocalization of anion transporter Slc26a7 in mouse kidney. AB - Previous studies have indicated that a major fraction of the filtered Cl(-) is reabsorbed via apical membrane Cl(-)/base exchange in the proximal tubule. Recent studies in Slc26a6 null mice have suggested that this transporter mediates only a portion of proximal tubule Cl(-)/base exchange, raising the possibility that one or more unidentified apical membrane transporters may additionally contribute. Recent studies have identified Slc26a7 as another Cl(-)/base exchanger expressed in the kidney. We therefore generated Slc26a7-specific polyclonal and monoclonal antibodies to examine cellular and subcellular sites of expression in mouse kidney. The specificity of each antibody was verified by immunoblotting and immunofluorescence of COS-7 cells transiently transfected with mouse Slc26a7. Immunofluorescence microscopy of mouse kidney detected the expression of Slc26a7 subapically in proximal tubule cells, and on the basolateral surface of thick ascending limb cells. Similar staining patterns were demonstrated with two antibodies shown to react with different epitopes on Slc26a7. Immunolocalization of Slc26a7 to proximal tubule and thick ascending limb was also observed in rat kidney. We conclude that Slc26a7 is expressed in the proximal tubule and thick ascending limb of the loop of Henle, and it may therefore contribute to anion transport in these nephron segments. PMID- 16263806 TI - Synergistic effects of PDGF-BB and cAMP-elevating agents on expression of connexin43 in mesangial cells. AB - The gap junction plays an important role in the regulation of cell growth, migration, and differentiation. Platelet-derived growth factor (PDGF) is reported to be a potent inhibitor of gap junctional intercellular communication (GJIC). Short-term exposure of cells to PDGF causes rapid and transient disruption of GJIC without altering connexin43 (Cx43) protein level. In this study, we investigated long-term effects of PDGF-BB on Cx43 expression in mesangial cells (MCs). Exposure of MCs to PDGF-BB affected neither the Cx43 protein level nor GJIC. However, in the presence of cAMP-elevating agents, PDGF-BB dramatically increased the expression of Cx43, which was accompanied by obviously augmented membrane distribution of Cx43 and functional GJIC. The increased expression of Cx43 was closely correlated with reduction in alpha-actin, a dedifferentiation marker of MCs. The effect of PDGF on Cx43 was largely prevented by inhibitors of phosphatidylinositol 3'-kinase or mitogen-activated protein kinase, but not by inhibition of protein kinase C. Exposure of MCs to PDGF-BB caused elevation in intracellular cAMP, and it was abolished by indomethacin, a cyclooxygenase inhibitor. However, indomethacin did not affect the synergistic effect. In addition, PDGF-BB also did not affect the degradation of Cx43. With the use of MCs transfected with a Cx43 promoter-luciferase vector, cooperative activation of Cx43 promoter by PDGF and cAMP was found. Together, our data reveal, for the first time, unexpected synergy between PDGF-BB and cAMP-elevating agents in the induction of Cx43 and MC differentiation. Regulation of GJIC could be an important mechanism via which PDGF modulates MC phenotypes. PMID- 16263807 TI - Cadmium inhibits both intrinsic and extrinsic apoptotic pathways in renal mesangial cells. AB - Cadmium is a potent nephrotoxin that has been shown to induce apoptosis in some cells but also to prevent it under certain circumstances. In several clinical situations and experimental models of injury to the renal glomerulus, pathological proliferation of mesangial cells is followed by resolution involving mesangial cell apoptosis. We investigated the effects of Cd(2+) on rat mesangial cells induced to undergo apoptosis through either the extrinsic receptor-mediated pathway or the intrinsic mitochondrial-dependent pathway. Camptothecin initiated the intrinsic pathway with activation of caspase-9 and caspase-dependent cleavage of procaspase-3. Tumor necrosis factor-alpha (TNF-alpha) initiated caspase-8 activity and cleavage of pro-caspase-3 at the convergence point of the two pathways. However, pro-caspase-8 levels were low, and caspase-9 was also activated in response to TNF-alpha, characteristic of what have been termed type II cells. With both TNF-alpha and camptothecin, concurrent exposure to 10 microM CdCl(2) suppressed DNA laddering, nuclear condensation, and pro-caspase-3 cleavage. It also decreased activity of both caspase-8 and caspase-9, prevented caspase-8-dependent cleavage of the proapoptotic factor Bid, and suppressed release of cytochrome c from mitochondria. At this 10-microM concentration, Cd(2+) was unique among a number of metal ions in preventing DNA fragmentation. We conclude that Cd(2+) is anti-apoptotic in rat mesangial cells, acting by a mechanism that may involve general caspase inhibition. This may have consequences for the resolution of nephritis in situations of mesangial cell hyperproliferation. PMID- 16263808 TI - Effects of ACE inhibition on proximal tubule sodium transport. AB - Angiotensin-converting enzyme (ACE) inhibitors such as captopril, which block ANG II formation, are commonly used for treatment of hypertension. There is substantial evidence that the proximal tubule (PT) is a primary target site for captopril but the molecular mechanisms for its action in PT are not well defined. The aim of this study was to determine the physiological and molecular changes in PT provoked by acute captopril treatment in the absence of changes in blood pressure or glomerular filtration rate (GFR). Captopril (infused at 12 microg/min for 20 min) did not change blood pressure or GFR but induced an immediate (<10 min) increase in PT flow measured with a nonobstructive optical method (to 117 +/ 14% of baseline) along with a rapid diuresis from 2.1 +/- 0.6 mg/min (baseline) to 3.7 +/- 0.9 mg/min (captopril). Captopril also provoked a significant retraction of PT Na(+)/H(+) exchanger isoform 3 (NHE3), NHE regulatory factor (NHERF)-1, myosin-VI, and Na(+)-P(i) cotransporter type 2 (NaPi2), but not ACE, out of apical microvillus-enriched membranes. Proteomic analysis with MALDI-TOF MS revealed an additional eight abundant membrane-associated proteins that redistributed out of the microvillus-enriched membrane during captopril treatment: megalin, myosin II-A, clathrin, aminopeptidase N, DPPIV, ezrin, moesin, and vacuolar H(+)-ATPase subunit beta(2). In summary, captopril can rapidly depress PT reabsorption in the absence of a change in GFR or BP and provokes the redistribution of a set of transporters and transporter-associated proteins that likely participate in the decrease in PT reabsorption and may also contribute to the blood pressure-lowering effect of ACE inhibitors. PMID- 16263809 TI - Coordinate control of prostaglandin E2 synthesis and uptake by hyperosmolarity in renal medullary interstitial cells. AB - During water deprivation, prostaglandin E(2) (PGE(2)), formed by renal medullary interstitial cells (RMICs), feedback inhibits the actions of antidiuretic hormone. Interstitial PGE(2) concentrations represent the net of both PGE(2) synthesis by cyclooxygenase (COX) and PGE(2) uptake by carriers such as PGT. We used cultured RMICs to examine the effects of hyperosmolarity on both PG synthesis and PG uptake in the same RMIC. RMICs expressed endogenous PGT as assessed by mRNA and immunoblotting. RMICs rapidly took up [(3)H]PGE(2) to a level 5- to 10-fold above background and with a characteristic time-dependent "overshoot." Inhibitory constants (K(i)) for various PGs and PGT inhibitors were similar between RMICs and the cloned rat PGT. Increasing extracellular hyperosmolarity to the range of 335-485 mosM increased the net release of PGE(2) by RMICs, an effect that was concentration dependent, maximal by 24 h, reversible, and associated with increased expression of COX-2. Over the same time period, there was decreased cell-surface activity of PGT due to internalization of the transporter. With continued exposure to hyperosmolarity over 7-10 days, PGE(2) release remained elevated, COX-2 returned to baseline, and PGT-mediated uptake became markedly reduced. Our findings suggest that hyperosmolarity induces coordinated changes in COX-2-mediated PGE(2) synthesis and PGT-mediated PGE(2) uptake in RMICs. PMID- 16263810 TI - Hypercalcemia of malignancy due to ectopic transactivation of the parathyroid hormone gene. AB - CONTEXT: The physiology of PTH is well described, but regulation of PTH gene expression remains enigmatic. This is, at least in part, because of a lack of suitable cell culture systems. OBJECTIVE, DESIGN, SETTING, PATIENTS, INTERVENTIONS, AND MAIN OUTCOME MEASURES: We report a case of severe hyperparathyroidism resulting from the ectopic production of PTH by a pancreatic malignancy. Cells from the primary tumor (PEPP1 cells) were established in culture to examine the etiology of ectopic PTH gene expression in this patient. RESULTS AND CONCLUSIONS: We failed to find amplification or rearrangement of the PTH gene but documented hypomethylation of the PTH promoter in tumor tissue. We found that PEPP1 cells support expression of a reporter gene containing regulatory sequences from the human PTH gene promoter. Therefore, this is the first report documenting ectopic PTH production by a tumor as the result of transactivation of the PTH gene. PEPP1 cells may be useful for future studies aimed at elucidating the details of PTH gene regulation. PMID- 16263811 TI - Identification of a functional polymorphism of the human type 5 17beta hydroxysteroid dehydrogenase gene associated with polycystic ovary syndrome. AB - CONTEXT: Polycystic ovary syndrome (PCOS) is characterized by chronic hyperandrogenic anovulation and is associated with insulin resistance. Its pathogenesis is believed to be multifactorial, and abnormal gene regulation could be one contributing factor. Type 5 17beta-hydroxysteroid dehydrogenase (17beta HSD5) appears to be the major testosterone-forming 17beta-HSD isoenzyme in females. OBJECTIVE: Our objective was to investigate the role of a potentially activating 17beta-HSD5 gene (HSD17B5) variant in hyperandrogenism. DESIGN AND SETTING: We conducted a case study and case-control cohort study in our General Clinical Research Center. STUDY SUBJECTS: Subjects included a case of PCOS who had hyperthecosis associated with profound type B insulin resistance and an unusual, frankly male testosterone response to a GnRH agonist test, as well as 121 PCOS patients and 128 population controls. INTERVENTIONS: Interventions were diagnostic. MAIN OUTCOME MEASURES: Main outcome measures included sequencing of HSD17B5 5'-flanking region and nine exons, genotype/phenotype studies, and in vitro functional studies. RESULTS: Our case had a previously undescribed homozygous HSD17B5 variant (G-to-A substitution) -71 bp in the promoter region. Genotyping controls showed this to be a single-nucleotide polymorphism (SNP)-71G. Luciferase activity of a SNP-71G promoter construct was significantly higher than that of the wild type, and EMSAs revealed that SNP-71G possessed significantly increased affinity to nuclear transcription factors. SNP-71G allele frequency (32.2 vs. 22.3%) and SNP-71G allele presence (53.7% vs. 38.3%) were significantly increased in PCOS (P = 0.01) [corrected] SNP-71G homozygosity tended to contribute about 20% to the plasma testosterone level. CONCLUSIONS: SNP-71G is a functional polymorphism that may contribute to testosterone excess in a subset of PCOS patients. PMID- 16263812 TI - Activity of irinotecan and the tyrosine kinase inhibitor CEP-751 in medullary thyroid cancer. AB - CONTEXT: Medullary thyroid cancer (MTC) is a cancer of the parafollicular C cells that commonly presents with an inherited or acquired RET gene mutation. There is currently no effective systemic treatment for MTC. OBJECTIVE: The objective of this study was to investigate a systemic therapeutic approach to treat MTC. We studied the sensitivity of an MTC cell line and xenograft to irinotecan, alone and in combination with the tyrosine kinase inhibitor, CEP-751. RESULTS: In TT cell culture and xenografts, irinotecan treatment was highly effective. This effect was augmented by treatment with CEP-751. Treatment of TT cell xenografts resulted in durable complete remission in 100% of the mice, with median time to recurrence of 70 d for irinotecan alone and more than 130 d for irinotecan plus CEP-751. Although irinotecan induced an S phase checkpoint arrest in TT cells, CEP-751 in combination with irinotecan resulted in a loss of this arrest. CEP-751 induced a loss in the induction of the DNA repair program marked by phospho-H2AX and the checkpoint pathway marked by the activated Chk1 pathway. CONCLUSIONS: Irinotecan treatment was highly effective in a preclinical model of human MTC, resulting in complete remission in 100% of the xenografts treated. The duration of remission was further enhanced by combination with the kinase inhibitor, CEP 751. These results suggest that irinotecan, alone or in combination, may be useful for the treatment of MTC. PMID- 16263813 TI - Detection of serum deoxyribonucleic acid methylation markers: a novel diagnostic tool for thyroid cancer. AB - CONTEXT: Serum DNA methylation markers may potentially be useful in diagnosing thyroid cancer and monitoring its recurrence. OBJECTIVE: The objective of the study was to assess the utility of serum DNA methylation as a diagnostic test for patients with thyroid nodules and a monitoring test to detect thyroid cancer recurrence in previously treated patients. DESIGN, SETTING, AND SUBJECTS: Using real-time quantitative methylation-specific PCR, we analyzed the methylation status of five genes (CALCA, CDH1, TIMP3, DAPK, and RARbeta2) on 96 bisulfite treated serum DNA samples isolated preoperatively from either solid thyroid nodule patients or patients in follow-up for history of treated thyroid cancer. MAIN OUTCOME MEASURE: Diagnostic sensitivity, specificity, and accuracy of serum DNA methylation marker for thyroid cancer were measured. RESULTS: For the patients with thyroid nodules, when a positive result was defined by a serum methylation level above the appropriately chosen cutoff value for any one of the five genes, the preoperative diagnostic sensitivity for thyroid cancer was 68% (26 of 38), the specificity was 95% (18 of 19), and the overall preoperative diagnostic accuracy was 77%, with positive and negative predictive values of 96 and 60%, respectively. In a subset of patients with cytologically indeterminate thyroid nodules, serum DNA methylation testing could correctly diagnose eight of 11 (73%) cancers and four of four (100%) benign tumors, with a diagnostic accuracy of 80%. We also analyzed these serum DNA methylation markers in 39 previously treated thyroid cancer patients. Among the 10 patients proved to have recurrent disease by conventional measures, seven (70%) were positive on methylation testing. Among the 29 patients who had no corroboration of residual or recurrent disease by conventional studies, six (21%) were positive for serum DNA methylation markers. CONCLUSIONS: We have demonstrated the potential usefulness of serum DNA methylation markers as a novel tool for differential diagnosis of solid thyroid nodules and thyroid cancer recurrence monitoring. PMID- 16263814 TI - Commentary: polycystic ovary syndrome: a syndrome of ovarian hypersensitivity to insulin? PMID- 16263815 TI - Neuropsychological function and symptoms in subjects with subclinical hypothyroidism and the effect of thyroxine treatment. AB - OBJECTIVE: Our objective was to examine the relation between neuropsychological function and subclinical hypothyroidism (SHT), defined as serum TSH of 3.5-10.0 mIU/liter and normal serum free T4 and free T3 levels, and to study the effect of T4 supplementation. SUBJECTS: A total of 89 subjects (45 males) with SHT and 154 control subjects (72 males) were recruited from a general health survey (the fifth Tromso study). Sixty-nine of those with SHT were included in a placebo controlled, double-blind intervention study with T4 medication for 1 yr. MAIN OUTCOME MEASURES: We used fourteen tests of cognitive function, Beck Depression Inventory, General Health Questionnaire, and a questionnaire on hypothyroid symptoms. RESULTS: The mean +/- sd serum TSH in the SHT and control group were 5.57 +/- 1.68 and 1.79 +/- 0.69 mIU/liter, respectively. There were no significant differences in cognitive function and hypothyroid symptoms between the two groups, but those with SHT scored significantly better than the controls on the GHQ-30. At the end of the intervention study, serum TSH in the T4 group (n = 36) and the placebo group (n = 33) were 1.52 +/- 1.51 and 5.42 +/- 1.96 mIU/liter, respectively. T4 substitution had no effect on any of the parameters measured. CONCLUSION: In subjects with SHT where the serum TSH level is in the 3.5-10.0 mIU/liter range, there is no neuropsychological dysfunction, and compared with healthy controls, there is no difference in symptoms related to hypothyroidism. PMID- 16263816 TI - Comparison between six-year therapy with long-acting somatostatin analogs and successful surgery in acromegaly: effects on cardiovascular risk factors. AB - CONTEXT: The effects of chronic therapy with long-acting somatostatin analogs (SSTa) on metabolic and cardiovascular parameters have been partially elucidated in acromegalic patients. OBJECTIVE: The objective of this study was to compare the long-term effects of SSTa treatment and successful surgery on GH/IGF-I secretion and cardiovascular risk parameters in acromegaly. DESIGN, PATIENTS, AND INTERVENTION: This was a retrospective study of 36 acromegalic patients treated with SSTa and evaluated after a median of 66 months and of 33 sex-, age-, and body mass index-matched cured patients evaluated after a similar period of remission, all from the Institute of Endocrine Sciences (Milan, Italy). MAIN OUTCOME MEASURES: The main outcome measures were fasting and post-oral load glucose homeostasis, hemoglobin A(1c), insulin sensitivity and secretion by several indexes, lipid profile, and blood pressure. RESULTS: Fasting and areas under the glucose response curve rose in patients controlled (n = 29) and not controlled (n = 7) by SSTa, becoming higher than those in cured subjects. A 1% hemoglobin A(1c) increase was observed in all nondiabetic SSTa patients, but not in cured subjects. Basal insulin secretion and resistance, evaluated by homeostasis model assessment, decreased in all SSTa patients, whereas oral glucose tolerance test-derived insulin secretion and resistance, evaluated by insulinogenic index and oral glucose tolerance test-derived insulin secretion, improved only in SSTa-treated controlled patients. Triglycerides did not change during SSTa, whereas high-density lipoprotein cholesterol increased in SSTa treated controlled patients. At the last visit, the contemporary presence of at least three cardiovascular risk factors was more frequent in patients treated with SSTa than in cured subjects. CONCLUSIONS: SSTa therapy induces long-lasting disease control and improvement of insulin sensitivity and high-density lipoprotein cholesterol levels in responsive patients. The progressive glucose homeostasis alterations, observed independently from the degree of cure, suggest the need for glucose homeostasis and peripheral vascular complications monitoring during chronic SSTa treatment. PMID- 16263818 TI - Autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy syndrome with renal failure: impact of posttransplant immunosuppression on disease activity. AB - CONTEXT: Autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy (APECED) is a rare autosomal recessive disorder caused by mutations in the gene AIRE (autoimmune regulator). APECED affects mainly endocrine organs resulting in hypoparathyroidism, adrenocortical failure, diabetes mellitus, hypogonadism, and hypothyroidism. Nonendocrine organ manifestations are autoimmune hepatitis, vitiligo, pernicious anemia, exocrine pancreatic insufficiency, and alopecia. APECED's first manifestation generally is mucocutaneous candidiasis presumably related to T cell dysfunction. PATIENT: A 5-yr-old Iranian girl presented first with pernicious anemia, exocrine pancreatic insufficiency, and nail candidiasis. She had renal dysfunction due to chronic interstitial nephritis (CIN), which progressed to end-stage renal failure. She was transplanted 1 yr later. Common causes of CIN were excluded. APECED was suspected first because she developed progressively hypoparathyroidism, adrenocortical failure, glucose intolerance, and hypothyroidism. RESULTS: Genetic analysis revealed a large homozygous deletion (g.424_2157del1734), spanning exons 2-4, in the AIRE gene. The predicted protein, if it is produced, has only 44 amino acids (exon 1) in common with the wild-type protein. Immunosuppression after the first renal transplant included prednisone, azathioprine, and cyclosporine A. Multiple acute rejection episodes occurred. Chronic rejection resulted in lost graft and she was retransplanted 2 yr later. Surprisingly, all APECED-related symptoms including candidiasis and autoantibody levels decreased, presumably due to the reinforced immunosuppression (tacrolimus, mycophenolate mofetil, prednisone). CONCLUSIONS: This is the first report of an APECED patient with CIN resulting in end-stage renal failure. Clinical and biological improvement was observed under posttransplant multidrug immunosuppression including tacrolimus and mycophenolate mofetil. PMID- 16263817 TI - Pharmacokinetics of a testosterone gel in healthy postmenopausal women. AB - BACKGROUND: The paucity of pharmacokinetic data on androgen formulations in women has hindered clinical trials of testosterone supplementation in women. OBJECTIVE: The objective of this study was to determine the time course and profile of serum testosterone concentrations during treatment with different doses of testosterone gel in postmenopausal women and assess whether estrogen treatment affects the pharmacokinetics of testosterone gel. METHODS: Postmenopausal women with total testosterone levels less than 33 ng/dl after baseline 24-h sampling were treated with 4.4, 8.8, or 13.2 mg testosterone gel daily for 7 d each in random order, with a 7-d washout period between doses. We studied 13 women who had not received estrogen therapy (group I) and 13 who had received stable estrogen therapy for 3 months or more (group II). Total and free testosterone concentrations were measured for 48 h on the seventh day of each dose administration. RESULTS: Twenty six women were randomized; of these, 24 were evaluable, 13 in group I and 11 in group II. The average steady-state concentrations (Cav) of serum total and free testosterone increased with increasing testosterone dose and were highly correlated with the dose (dose effect, P < 0.00001), but were not affected by estrogen therapy (P = 0.43). In both groups, the 4.4-mg dose increased Cav total and free testosterone into the mid- to high-normal range, whereas 8.8- and 13.2 mg doses raised total (Cav: 22.3, 51.6, 80.3, and 92.0 ng/dl in group I; 22.7, 59.8, 82.0, and 114.3 ng/dl in group II at 0, 4.4, 8.8, and 13. 2 mg, respectively) and free testosterone (5.9, 8.4, 11.5,12.8 pg/ml in group I and 5.0,7.6,11.1,10.8 in group II, respectively, at the various doses) above the physiological range. The area under the curve, maximum and minimum concentrations, and the change in Cav for total and free testosterone were dose related and significantly higher during administration of the 13.2-mg dose than during the 0- or 4.4-mg dose; estrogen therapy had no significant effect on these measures. Serum estradiol, LH, FSH, and SHBG levels did not change significantly at any dose. Testosterone gel was well tolerated. CONCLUSIONS: Administration of testosterone gel to postmenopausal women raised total and free testosterone concentrations in proportion to the administered dose without affecting estradiol levels. A 4.4-mg dose raised testosterone levels into the mid- to high-normal range. Previous estrogen therapy had no significant effect on testosterone pharmacokinetics over this short duration. PMID- 16263819 TI - Sympathovagal imbalance in acromegalic patients. AB - CONTEXT: Sympathovagal imbalance is a common finding in diabetes and is considered to be a cardiovascular risk factor. No data are available on sympathovagal balance (SB) in acromegalic patients. OBJECTIVE: The objective of this study was to evaluate SB in acromegalic patients. PATIENTS: Twenty nondiabetic, nonhypopituitary, acromegalic patients (13 women and seven men; mean age +/- sem, 51.30 +/- 3.09 yr) were compared with age-matched subjects (21 normal subjects, 20 patients with type 1 diabetes mellitus, and 15 patients with type 2 diabetes mellitus). INTERVENTIONS: Autonomic tests, used to evaluate SB, were performed by power spectral analysis of heart rate variability in clinostatism (c) and orthostatism (o), using a frequency domain method. Power spectral analysis identifies peaks of power: high frequency (HF), which expresses vagal activity, and low frequency (LF), which expresses sympathetic activity. RESULTS: Acromegalic patients displayed significantly lower LFc/HFc (P = 0.002) and LFo/HFo (P < 0.001) ratios than normal subjects. HFo was significantly higher in acromegalic patients than in normal subjects (P < 0.001) and patients with type 1 diabetes mellitus (P = 0.004), but no different from that in type 2 diabetes mellitus patients (P = 0.069). In untreated acromegalic patients, the alterations found in the whole group were confirmed; no statistically significant differences were found between untreated acromegalic patients and those treated with somatostatin analogs. Similarly, the same alterations found in the whole group were evident in the controlled acromegalic patients, and no significant differences were found between controlled and uncontrolled patients. CONCLUSION: Our study evidenced that sympathovagal imbalance in acromegalic patients, due to vagal hypertone, is difficult to reverse and is not influenced by medical therapy. This could be a new cardiovascular risk factor. PMID- 16263820 TI - Extensive clinical experience: relative prevalence of different androgen excess disorders in 950 women referred because of clinical hyperandrogenism. AB - CONTEXT: We undertook this study to estimate the prevalence of the various androgen excess disorders using the new criteria suggested for the diagnosis of polycystic ovary syndrome (PCOS). SETTING: The study was performed at two endocrine departments at the University of Palermo (Palermo, Italy). PATIENTS: The records of all patients referred between 1980 and 2004 for evaluation of clinical hyperandrogenism were reevaluated. All past diagnoses were reviewed using the actual diagnostic criteria. To be included in this study, the records of the patients had to present the following available data: clinical evaluation of hyperandrogenism, body weight and height, testosterone (T), free T, dehydroepiandrosterone sulfate, 17-hydroxyprogesterone, progesterone, and pelvic sonography. A total of 1226 consecutive patients were seen during the study period, but only the scores of 950 patients satisfied all criteria and were reassessed for the diagnosis. RESULTS: The prevalence of androgen excess disorders was: PCOS, 72.1% (classic anovulatory patients, 56.6%; mild ovulatory patients, 15.5%), idiopathic hyperandrogenism, 15.8%; idiopathic hirsutism, 7.6%; 21-hydroxylase-deficient nonclassic adrenal hyperplasia, 4.3%; and androgen secreting tumors, 0.2%. Compared with other androgen excess disorders, patients with PCOS had increased body weight whereas nonclassic adrenal hyperplasia patients were younger and more hirsute and had higher serum levels of T, free T, and 17-hydroxyprogesterone. CONCLUSIONS: Classic PCOS is the most common androgen excess disorder. However, mild androgen excess disorders (ovulatory PCOS and idiopathic hyperandrogenism) are also common and, in an endocrine setting, include about 30% of patients with clinical hyperandrogenism. PMID- 16263822 TI - Survival and death causes in differentiated thyroid carcinoma. AB - BACKGROUND: Survival studies in differentiated thyroid carcinoma (DTC) may be biased because they have been performed in heterogeneous populations. In addition, specific death causes in DTC have not been documented well in the literature. AIMS: The aim of our study was to investigate survival and specific death causes in a homogeneous cohort of DTC patients. PATIENTS: Patients included 366 consecutive patients with DTC who had all been treated according to the same protocol for initial therapy and follow-up. METHODS: Prognostic factors for DTC related death were analyzed by univariate Cox regression analysis, followed by stepwise multivariate Cox regression analysis. Standardized mortality rates (SMR) were calculated using normal mortality rates for the entire Dutch population. RESULTS: During follow-up of 8.3 +/- 4.6 yr, 82 patients (22.4%) died. At multivariate Cox-regression analysis, tumor stage T4, distant metastases, and advanced age were associated with an increased relative risk for DTC-related death. SMR for the entire group was 2.32. This could be explained by increased SMR in patients with stage T4, distant metastases, or advanced age. Death causes could be verified in 80 patients: 52 died of DTC, 28 due to other causes. Ten of the 20 patients with stage T1-3M0 died from thyroid carcinoma. CONCLUSION: Relative risk for thyroid cancer-related death and SMR are significantly increased in patients with stage T4 and M1 or advanced age. Although death risk is not increased in T1-3 M0 patients, DTC contributed significantly to mortality in all patient categories. PMID- 16263821 TI - Wnt/beta-catenin signaling mediates antineoplastic effects of imatinib mesylate (gleevec) in anaplastic thyroid cancer. AB - CONTEXT: Dysregulation of Wnt signaling is a key step in neoplastic thyrocyte proliferation. However, it is unclear whether the selective tyrosine kinase (TK) inhibitor, imatinib mesylate, is linked to the Wnt/beta-catenin cascade and is able to modulate the pathway. OBJECTIVE: Conflicting data are reported on the therapeutic effects of imatinib in anaplastic thyroid carcinomas (ATCs), but the molecular mechanism of action is unclear. Here, we further delineated the antitumor effects and the potential efficacy of imatinib in dedifferentiated thyroid carcinomas. RESULTS: Tissue microarray of histologically proven ATCs (n = 12) demonstrated that six of 12 tumors expressed at least one of the imatinib sensitive TKs. Similarily, imatinib-sensitive TKs were detected in seven of 10 thyroid cancer cell lines derived from metastatic papillary, follicular, and ATCs. Coimmunoprecipitation in ARO cells demonstrated a direct link between c-abl and beta-catenin. Imatinib (10 microM for 48 h) drastically reduced beta-catenin expression and redistributed it from the nucleus to the cell membrane. It stabilized adherens junctions by increasing beta-catenin/E-cadherin binding and reduced the invasive potential of thyroid cancer. Furthermore, imatinib (10 microM for 48 h) attenuated T cell factor/lymphoid enhancer factor activity, reduced cyclin D1 levels and dose-dependently suppressed thyrocyte proliferation by half without affecting apoptosis. CONCLUSION: Our data provide a molecular mechanism for the antitumor activity of imatinib that may help to develop it as a therapeutic option in a subset of ATC patients. PMID- 16263823 TI - Helicobacter pylori infection and autoimmune thyroid disease in young patients: the disadvantage of carrying the human leukocyte antigen-DRB1*0301 allele. AB - CONTEXT AND OBJECTIVE: Pathogenesis of autoimmune thyroid disease (ATD) is multifactorial. Helicobacter pylori (Hp) infection has been proposed to be involved in nongastrointestinal conditions and reported more frequently in ATD adult patients. We evaluated the prevalence of Hp antibodies in young ATD patients and investigated the possibility that a susceptible immunogenetic profile could influence the development of ATD in subjects with Hp infection. SUBJECTS AND METHODS: We retrospectively studied 90 children with ATD (median age 11.2 yr), 70 age- and sex-matched healthy subjects as controls, and 65 patients with Turner syndrome (median age 18.8 yr). Antibodies to Hp were determined at diagnosis in ATD patients and, in Turner patients, at the last control in cases without ATD and before the appearance of thyroid autoantibodies in the others. Serological and molecular human leukocyte antigen (HLA) typing for classes I and II polymorphisms was performed. RESULTS: Prevalence of positive Hp serology resulted significantly higher in ATD patients than controls (P = 0.032). No association was found between individual HLA alleles and Hp serology. HLA-A1, B8, and DRB1*0301 were found significantly associated with ATD. A significant interaction between HLA-DRB1*0301 and Hp infection was present in ATD patients and not controls (P = 0.007), suggesting that the copresence of these two factors might favor ATD development. A similar phenomenon was observed in Turner syndrome patients (P = 0.02; cumulative Mantel test, P = 0.0001). CONCLUSIONS: Another target of Hp-elicited immune inflammatory response might be the thyroid gland in subjects with a peculiar immunogenetic profile so that ATD may be a consequence. Our findings suggest the opportunity of eradicating Hp infection in children with ATD and/or susceptible HLA alleles. PMID- 16263824 TI - The role of CBP/p300 interactions and Pit-1 dimerization in the pathophysiological mechanism of combined pituitary hormone deficiency. AB - CONTEXT: Combined pituitary hormone deficiency (CPHD) in humans is caused by mutations of pituitary-specific transcription factors such as Pit-1. Although many patients with CPHD have an autosomal recessive disorder caused by a Pit-1 DNA-binding mutation, there are a number of reports of mutant Pit-1 molecules that either by prediction or through experimentation bind normally to DNA. OBJECTIVE: The objective of this study was to understand the pathophysiological mechanisms of mutant Pit-1 molecules with intact DNA binding. DESIGN: DNA-binding and functional studies were used to assess five Pit-1 mutations: F135C, R143Q, A158P, K216E, and R271W. RESULTS: In gel-shift studies using well-characterized DNA-binding elements from the GH and prolactin genes, the K126E mutant displayed markedly enhanced Pit-1 dimer binding to either element, whereas the R271W mutant bound with high avidity, but only as a monomer. In contrast, the R143Q mutant was unable to bind these elements, and the F135C and A158P mutants displayed near normal DNA-binding characteristics. We observed that CBP/p300 bound poorly to the A158P and K216E mutant Pit-1 molecules, but bound normally to the F135C, R143Q, and R271W mutants. In functional assays, CBP/p300 cotransfection with mutant Pit 1 expression vectors resulted in less transactivation of either the GH or prolactin reporter genes. CONCLUSIONS: From these studies, we suggest that CBP/p300 recruitment and Pit-1 dimerization are necessary for Pit-1 target gene activation and are important in the pathogenesis of CPHD. PMID- 16263825 TI - Correlation between testosterone and the inflammatory marker soluble interleukin 6 receptor in older men. AB - CONTEXT: An age-associated decline in testosterone (T) levels and an increase in proinflammatory cytokines contribute to chronic diseases in older men. Whether and how these changes are related is unclear. OBJECTIVE: We hypothesized that T and inflammatory markers are negatively correlated in older men. DESIGN: This was a cross-sectional study. SETTING: A population-based sample of older men was studied. PARTICIPANTS AND MEASURES: After excluding participants taking glucocorticoids or antibiotics or those with recent hospitalization, 467 men, aged 65 yr or older, had complete determinations of total T, bioavailable T, SHBG, albumin, IL-6, soluble IL-6 receptor (sIL-6r), TNF-alpha, IL-1beta, and C reactive protein. RESULTS: After adjusting for potential confounders, sIL-6r was significantly and inversely correlated with total T (r = -0.20; P < 0.001) and bioavailable T (r = -0.12; P < 0.05). T was not correlated with any other inflammatory marker. CONCLUSIONS: These preliminary findings suggest an inverse relationship between T and sIL-6r. Longitudinal studies are needed to establish the causality of this association. PMID- 16263826 TI - Maternal serum-soluble vascular endothelial growth factor receptor-1 in early pregnancy ending in preeclampsia or intrauterine growth retardation. AB - CONTEXT: Vascular endothelial growth factor (VEGF) promotes placental vascularization, which is inadequate in preeclampsia and intrauterine growth retardation (IUGR). The soluble receptor of VEGF (sVEGFR-1), also known as soluble fms-like tyrosine kinase-1, is produced in the placenta and reduces VEGF activity. Therefore, elevated sVEGFR-1 could contribute to the development of preeclampsia and IUGR. OBJECTIVE: The objective of this study was to study maternal serum sVEGFR-1 concentration in early pregnancy ending in preeclampsia and IUGR. DESIGN: This was a case-control study. SETTING: This study was conducted at Helsinki University Central Hospital (Helsinki, Finland), a tertiary referral center. PATIENTS: Patients included 124 pregnant women, of whom 49 developed preeclampsia, 16 gave birth to IUGR infants without preeclampsia, and 59 remained normotensive and gave birth to normal-sized infants. Serum samples were collected at 12-15 and 16-20 gestational weeks. MAIN OUTCOME MEASURES: Serum sVEGFR-1 concentrations were determined by ELISA. RESULTS: Women with subsequent preeclampsia had higher [median; interquartile range (IQR)] concentrations of sVEGFR-1 at 16-20 wk gestation (436 and 282-699 ng/liter; P = 0.005) than the controls (296 and 184-508 ng/liter). The conclusion was the same if women with mild (340 and 285-750 ng/liter; P = 0.043) or severe (497 and 235-699 ng/liter; P = 0.022) preeclampsia were analyzed separately. An elevated sVEGFR-1 concentration at 16-20 wk gestation is associated with an increased risk of preeclampsia but not of isolated IUGR. Soluble VEGFR-1 concentration decreased by 15% from the first to the second sampling in the controls but not in women with preeclampsia or IUGR. CONCLUSION: Elevated sVEGFR-1 concentrations at 16-20 wk gestation precede the clinical manifestations of preeclampsia. By neutralizing VEGF, sVEGFR-1 may contribute to inadequate placental vascularization. PMID- 16263828 TI - Effect of growth hormone dose on bone maturation and puberty in children with idiopathic short stature. AB - CONTEXT: GH at 0.22 mg/kg.wk has been shown to have no effect on pubertal onset or pace, whereas GH at 0.5 mg/kg x wk has been shown to advance pubertal onset and bone maturation. OBJECTIVES: Our objectives were to determine whether 0.37 mg/kg x wk GH advanced pubertal onset, pace, or bone maturation relative to 0.24 mg/kg x wk GH; whether 0.37 mg/kg x wk GH led to pubertal onset at an inappropriately early age; and whether age at start of GH therapy influenced pubertal onset. DESIGN: We conducted a randomized, open-label study to final height. PATIENTS: We studied children with idiopathic short stature. INTERVENTION: Patients were treated with 0.24 mg/kg x wk, 0.24 increasing to 0.37 mg/kg x wk, or 0.37 mg/kg x wk. MAIN OUTCOME MEASURES: We assessed age at pubertal onset and rates of bone maturation, Tanner stage development, and increase in testicular volume (boys only). RESULTS: For the primary comparison between the 0.24 and 0.37 mg/kg x wk dose groups, median ages of pubertal onset (in years) were similar (13.7 vs. 13.5 for boys and 11.7 vs. 11.4 for girls) and were greater than those for the general population for each sex. Age at start of GH therapy did not appear to influence pubertal onset for either sex. Rates of pubertal pace and bone maturation were not significantly different between the 0.24 and 0.37 mg/kg x wk dose groups for either sex. CONCLUSION: GH at 0.37 mg/kg x wk does not appear to accelerate pubertal onset, pace, or bone maturation compared with GH at 0.24 mg/kg x wk in patients with idiopathic short stature. From a clinical standpoint, our results suggest that the approved dose range of up to 0.37 mg/kg x wk GH does not lead to pubertal onset at an inappropriately early age. PMID- 16263827 TI - Advancing human circadian rhythms with afternoon melatonin and morning intermittent bright light. AB - CONTEXT: Both light and melatonin can be used to phase shift the human circadian clock, but the phase-advancing effect of the combination has not been extensively investigated. OBJECTIVE: The objective of the study was to determine whether phase advances induced by morning intermittent bright light and a gradually advancing sleep schedule could be increased with afternoon melatonin. PARTICIPANTS: Healthy adults (25 males, 19 females, between the ages of 19 and 45 yr) participated in the study. DESIGN: There were 3 d of a gradually advancing sleep/dark period (wake time 1 h earlier each morning), bright light on awakening [four 30-min bright-light pulses (approximately 5000 lux) alternating with 30 min room light < 60 lux] and afternoon melatonin, either 0.5 or 3.0 mg melatonin timed to induce maximal phase advances, or matching placebo. The dim light melatonin onset was measured before and after the treatment to determine the phase advance. RESULTS: There were significantly larger phase advances with 0.5 mg (2.5 h, n = 16) and 3.0 mg melatonin (2.6 h, n = 13), compared with placebo (1.7 h, n = 15), but there was no difference between the two melatonin doses. Subjects did not experience jet lag-type symptoms during the 3-d treatment CONCLUSIONS: Afternoon melatonin, morning intermittent bright light, and a gradually advancing sleep schedule advanced circadian rhythms almost 1 h/d and thus produced very little circadian misalignment. This treatment could be used in any situation in which people need to phase advance their circadian clock, such as before eastward jet travel or for delayed sleep phase syndrome. PMID- 16263829 TI - Nuclear receptor coactivator-3 alleles are associated with serum bioavailable testosterone, insulin-like growth factor-1, and vertebral bone mass in men. AB - CONTEXT: Nuclear receptor coactivator-3 (NCOA3) is a member of the steroid receptor coactivator family that interacts with nuclear hormone receptors to enhance their transcriptional activation function and may play a role in somatic growth. OBJECTIVE: The aim of this study was to examine the relationships between the CAG/CAA (glutamine) length variation at the NCOA3 locus, sex steroid hormone, and IGF-I levels and bone mineral density (BMD) in a cohort of older Caucasian men. DESIGN AND METHODS: We analyzed the association between potentially functional alleles at this locus, serum sex steroid hormone, and IGF-I levels and lumbar spine and proximal femur BMD (Hologic QDR) in 263 community-dwelling Caucasian men (age 66 +/- 7 yr, mean +/- sd; range 51-84 yr). Men were genotyped for a CAG/CAA repeat polymorphism in NCOA3, which encodes a polyglutamine tract of variable length in the C-terminal transcriptional activation domain of the protein. RESULTS: We found a significant monotonic decrease in lumbar spine, but not hip, BMD with increasing copies of the most common allele (29 repeats, 53%). For example, men with the 29/29 genotype had 6% or nearly 0.5 sd lower spine BMD than men without this genotype, and NCOA3 genotype explained 3.2% of the phenotypic variation in this trait. Serum levels of bioavailable testosterone and IGF-I paralleled genotype-related differences in lumbar spine BMD. CONCLUSION: Allelic variation at the NCOA3 locus may contribute to the genetic control of androgenic hormone and IGF levels and vertebral bone mass among older men. PMID- 16263830 TI - Beta-cell function, insulin sensitivity, and glucose tolerance in obese diabetic and nondiabetic adolescents and young adults. AB - CONTEXT: Type 2 diabetes mellitus (T2DM) is estimated to account for 10-45% of incident pediatric diabetes cases. OBJECTIVE: Our objective was to characterize the metabolic defects underlying T2DM in adolescents and young adults. DESIGN, SETTING, AND PATIENTS: We conducted a cross-sectional study of islet function and insulin sensitivity in 16 adolescents with T2DM and 13 obese (OB) and 13 lean (LN) age-matched nondiabetic subjects at a University Medical Center. INTERVENTION: We provided oral and iv glucose tolerance tests. MAIN OUTCOME MEASURES: We measured insulin and glucagon levels, insulin sensitivity, acute insulin responses to iv glucose, and the ratio of proinsulin to immunoreactive insulin. RESULTS: The diabetic subjects had elevated fasting insulin levels and significantly reduced insulin sensitivity (P < 0.05). The acute insulin response to iv glucose was comparable in the T2DM and LN groups (P < 0.05 for the OB vs. LN and T2DM), but insulin secretion adjusted for insulin resistance, the disposition index, was severely impaired in the diabetic subjects (P < 0.05 for the T2DM vs. LN and OB). The ratio of proinsulin to immunoreactive insulin did not differ among the three groups in the basal or stimulated state. Plasma glucagon levels were comparable before and after ingestion of glucose. CONCLUSIONS: These findings demonstrate that diabetic adolescents have significant insulin resistance, even compared with subjects of similar obesity and body fatness, and impaired insulin secretion relative to their degree of insulin resistance. However, the adolescent diabetic subjects retained a first phase insulin response to glucose that was comparable to lean controls and did not have hyperproinsulinemia or hyperglucagonemia. PMID- 16263831 TI - Eotaxin and obesity. AB - CONTEXT: Asthma and obesity incidence is increasing worldwide, and asthma is often more severe in the obese. Eotaxin, a CC chemokine, is important in extrinsic asthma, an inflammatory disorder. OBJECTIVE: Our objective was to examine the relation between eotaxin and obesity. DESIGN: We conducted a comparison study of eotaxin in mice fed high-fat vs. standard chow diet for 26 wk, in obese vs. lean humans, in obese humans before and after 4-6 wk of weight loss, and in sc vs. visceral adipose tissue from patients undergoing bariatric surgery. SETTING: Our clinical study occurred in an outpatient weight loss program. PATIENTS: Patients were obese adults with metabolic syndrome (n = 40) and nine morbidly obese bariatric surgery patients. INTERVENTION: Intervention was a very-low-calorie diet. MAIN OUTCOME MEASURES: We assessed circulating eotaxin and eotaxin mRNA levels in adipose tissue. RESULTS: Serum eotaxin levels were significantly higher in obese mice, and adipose mRNA levels correlated positively with serum eotaxin levels. Adipose tissue explants from obese mice showed increased secretion of eotaxin compared with explants from lean mice. In obese patients, plasma eotaxin levels were significantly higher than in lean controls and significantly reduced after weight loss, and eotaxin mRNA levels were 4.7-fold higher in visceral than sc adipose tissue. CONCLUSIONS: Circulating eotaxin and eotaxin mRNA levels in visceral adipose tissue were increased in obesity in mice and humans. Adipose tissue explants secrete eotaxin, and the stromal/vascular component of adipose tissue seems to be the predominant source of eotaxin. Diet-induced weight loss in humans led to reduction in plasma eotaxin levels, demonstrating that clinical interventions that target obesity can modulate systemic eotaxin levels. PMID- 16263832 TI - Partial surgical removal of growth hormone-secreting pituitary tumors enhances the response to somatostatin analogs in acromegaly. AB - CONTEXT: Surgery is a cornerstone in the treatment of acromegaly, but its efficacy in large, invasive tumors is scant. OBJECTIVE: The objective of this study was to investigate whether partial surgical removal of GH-secreting pituitary tumors enhances the response rate to somatostatin analogs (SSA; sc octreotide, slow-release octreotide, and lanreotide). DESIGN: This was a multicenter, open, retrospective study. SETTING: The study was performed at university hospitals. SUBJECTS AND METHODS: Eighty-six patients (42 women and 44 men; age, 42 +/- 14 yr) with acromegaly were studied. INTERVENTIONS: Patients underwent two courses of octreotide, lanreotide, or slow-release octreotide treatments before and after surgery of at least 6 months. MAIN OUTCOME MEASURE: The main outcome measure was normal IGF-I levels for age. RESULTS: Presurgical SSA treatment significantly decreased GH and IGF-I levels in all patients. GH levels were less than 2.5 microg/liter in 12 patients (14%); IGF-I levels normalized in nine (10%). After surgery, GH and IGF-I levels further decreased in all patients; tumor removal was greater than 75% in 50 (58%), 50.1-75% in 21 (24%), 25.1-50% in 10 (12%), and less than 25% in five patients (6%). Preoperatively, pituitary function was impaired in 12 patients (14%). Postsurgical SSA treatment lowered GH levels to less than 2.5 microg/liter in 49 (56%) and normalized IGF-I levels in 48 patients (55%). The success rate was significantly increased compared with that before surgery (P < 0.0001). GH (r = 0.48; P < 0.0001) and IGF-I levels (r = -0.38; P = 0.0003) after postsurgery SSA treatment correlated with the amount of tumor surgically removed. After surgery, pituitary function was impaired in 28 patients (32.6%) and was improved in 12 patients (13.9%). The cumulative prevalence of pituitary deficiency did not change during the study (normal function from 40 to 42%; deficiency from 60 to 58%). CONCLUSIONS: Surgical tumor removal (>75%) enhances the response to SSAs without impairing pituitary function. Our data indicate that surgical debulking has a significant place in the treatment algorithm of acromegaly. PMID- 16263833 TI - Noonan syndrome: relationships between genotype, growth, and growth factors. AB - CONTEXT: Half of the patients with Noonan syndrome (NS) carry mutation of the PTPN11 gene, which plays a role in many hormonal signaling pathways. The mechanism of stunted growth in NS is not clear. OBJECTIVE: The objective of the study was to compare growth and hormonal growth factors before and during recombinant human GH therapy in patients with and without PTPN11 mutations (M+ and M-). SETTING, DESIGN, AND PATIENTS: This was a prospective multicenter study in 35 NS patients with growth retardation. Auxological data and growth before and during 2 yr of GH therapy are shown. GH, IGF-I, IGF binding protein (IGFBP)-3, and acid-labile subunit (ALS) levels were evaluated before and during therapy. RESULTS: Molecular investigation of the PTPN11 coding sequence revealed 12 different heterozygous missense mutations in 20 of 35 (57%). Birth length was reduced [mean -1.2 sd score (SDS); six m+ and two m- were < -2 SDS] but not birth weight. M+ vs. M- patients were shorter at 6 yr (P = 0.04). In the prepubertal group (n = 25), GH therapy resulted in a catch-up height SDS, which was lower after 2 yr in M+ vs. M- patients (P < 0.03). The mean peak GH level (n = 35) was 15.4 +/- 6.5 ng/ml. Mean blood IGF-I concentration in 19 patients (11 m+, eight m ) was low (especially in M+) for age, sex, and puberty (-1.6 +/- 1.0 SDS) and was normalized after 1 yr of GH therapy (P < 0.001), without difference in M+ vs. M- patients. ALS levels (n = 10) were also very low. By contrast, the mean basal IGFBP-3 value (n = 19) was normal. CONCLUSIONS: In NS patients with short stature, some neonates have birth length less than -2 SDS. Growth of M+ is reduced and responds less efficiently to GH than M- patients. The association of low IGF-I and ALS with normal IGFBP-3 levels could explain growth impairment of M+ children and could suggest a GH resistance by a late postreceptor signaling defect. PMID- 16263834 TI - Effects of high-dose growth hormone on glucose and glycerol metabolism at rest and during exercise in endurance-trained athletes. AB - CONTEXT: Recombinant human-GH (r-hGH), in supraphysiological doses, is self administered by athletes in the belief that it is performance enhancing. OBJECTIVE: The objective of this study was to determine whether r-hGH alters whole-body glucose and glycerol metabolism in endurance-trained athletes at rest and during and after exercise. DESIGN: This was a 4-wk double-blind placebo controlled trial. SETTING: This study was conducted at St. Thomas Hospital (London, UK). PARTICIPANTS: Twelve endurance-trained male athletes were recruited and randomized to r-hGH (0.2 U/kg.d) (n = 6) or identical placebo (n = 6) for 4 wk. One (placebo group) withdrew after randomization. INTERVENTION: Intervention was conducted by randomization to r-hGH (0.2 U/kg x d) or identical placebo for 4 wk. MAIN OUTCOME MEASURES: Whole-body rates of appearance (Ra) of glucose and glycerol (an index of lipolysis) and rate of disappearance of glucose were measured using infusions of d-[6-6-2H2]glucose and 2H5-glycerol. RESULTS: Plasma levels of glycerol and free fatty acids and glycerol Ra at rest and during and after exercise increased during r-hGH treatment (P < 0.05 vs. placebo). Glucose Ra and glucose rate of disappearance were greater after exercise during r-hGH treatment (P < 0.05 vs. placebo). Resting energy expenditure and fat oxidation were greater under resting conditions during r-hGH treatment (P < 0.05 vs. placebo). CONCLUSIONS: r-hGH in endurance-trained athletes increased lipolysis and fatty acid availability at rest and during and after exercise. r-hGH increased glucose production and uptake rates after exercise. The relevance of these effects for athletic performance is not known. PMID- 16263836 TI - The neuropsychiatry of aging. PMID- 16263835 TI - Septic shock and sepsis: a comparison of total and free plasma cortisol levels. AB - CONTEXT: Severe systemic infection leads to hypercortisolism. Reduced cortisol binding proteins may accentuate the free cortisol elevations seen in systemic infection. Recently, low total cortisol increments after tetracosactrin have been associated with increased mortality and hemodynamic responsiveness to exogenous hydrocortisone in septic shock (SS), a phenomenon termed by some investigators as relative adrenal insufficiency (RAI). HYPOTHESIS: Free plasma cortisol may correspond more closely to illness severity than total cortisol, comparing SS and sepsis (S). DESIGN: This was a prospective study. SETTING: This study took place in a tertiary teaching hospital. PATIENTS: Patients had SS (n = 45) or S (n = 19) or were healthy controls (HCs; n = 10). AIM: The aim of the study was to compare total with free cortisol, measured directly and estimated by Coolens' method, corticosteroid-binding globulin (CBG), and albumin in patients with SS (with and without RAI) and S during acute illness, recovery, and convalescence. RESULTS: Comparing SS, S, and HC subjects, free cortisol levels reflected illness severity more closely than total cortisol (basal free cortisol, SS, 186 vs. S, 29 vs. HC, 13 nmol/liter, P < 0.001 compared with basal total cortisol, SS, 880 vs. S, 417 vs. HC, 352 nmol/liter, P < 0.001). Stimulated free cortisol increments varied greatly with illness category (SS, 192 vs. S, 115 vs. HC, 59 nmol/liter, P = 0.004), whereas total cortisol increments did not (SS, 474 vs. S, 576 vs. HC, 524 nmol/liter, P = 0.013). The lack of increase in total cortisol with illness severity is due to lower CBG and albumin. One third of patients with SS (15 of 45) but no S patients met a recently described criterion for RAI (total cortisol increment after tetracosactrin < or = 248 nmol/liter). RAI patients had higher basal total cortisol (1157 vs. 756 nmol/liter; P = 0.028) and basal free cortisol (287 vs. 140 nmol/liter; P = 0.017) than non-RAI patients. Mean cortisol increments in RAI were lower (total, 99 vs. 648 nmol/liter, P < 0.001; free, 59 vs. 252 nmol/liter, P < 0.001). These differences were not due to altered CBG or albumin levels. Free cortisol levels normalized more promptly than total cortisol in convalescence. Calculated free cortisol by Coolens' method compared closely with measured free cortisol. CONCLUSIONS: Free cortisol is likely to be a better guide to cortisolemia in systemic infection because it corresponds more closely to illness severity. The attenuated cortisol increment after tetracosactrin in RAI is not due to low cortisol-binding proteins. Free cortisol levels can be determined reliably using total cortisol and CBG levels. PMID- 16263837 TI - Systematic review of psychological approaches to the management of neuropsychiatric symptoms of dementia. AB - OBJECTIVE: The authors systematically reviewed the literature on psychological approaches to treating the neuropsychiatric symptoms of dementia. METHOD: Reports of studies that examined effects of any therapy derived from a psychological approach that satisfied prespecified criteria were reviewed. Data were extracted, the quality of each study was rated, and an overall rating was given to each study by using the Oxford Centre for Evidence-Based Medicine criteria. RESULTS: A total of 1,632 studies were identified, and 162 satisfied the inclusion criteria for the review. Specific types of psychoeducation for caregivers about managing neuropsychiatric symptoms were effective treatments whose benefits lasted for months, but other caregiver interventions were not. Behavioral management techniques that are centered on individual patients' behavior or on caregiver behavior had similar benefits, as did cognitive stimulation. Music therapy and Snoezelen, and possibly sensory stimulation, were useful during the treatment session but had no longer-term effects; interventions that changed the visual environment looked promising, but more research is needed. CONCLUSIONS: Only behavior management therapies, specific types of caregiver and residential care staff education, and possibly cognitive stimulation appear to have lasting effectiveness for the management of dementia-associated neuropsychiatric symptoms. Lack of evidence regarding other therapies is not evidence of lack of efficacy. Conclusions are limited because of the paucity of high-quality research (only nine level-1 studies were identified). More high-quality investigation is needed. PMID- 16263838 TI - Epidemiology of and risk factors for psychosis of Alzheimer's disease: a review of 55 studies published from 1990 to 2003. AB - OBJECTIVE: The authors reviewed studies published between 1990 and 2003 that reported the prevalence, incidence, and persistence of, as well as the risk factors associated with, psychosis of Alzheimer's disease. METHOD: PubMed and PsycINFO databases were searched by using the terms "psychosis and Alzheimer disease" and "psychosis and dementia." Empirical investigations presenting quantitative data on the epidemiology of and/or risk factors for psychotic symptoms in Alzheimer's disease were included in the review. A total of 55 studies, including a total of 9,749 subjects, met the inclusion criteria. RESULTS: Psychosis was reported in 41% of patients with Alzheimer's disease, including delusions in 36% and hallucinations in 18%. The incidence of psychosis increased progressively over the first 3 years of observation, after which the incidence seemed to plateau. Psychotic symptoms tended to last for several months but became less prominent after 1 year. African American or black ethnicity and more severe cognitive impairment were associated with a higher rate of psychosis. Psychosis was also associated with more rapid cognitive decline. Some studies found a significant association between psychosis and age, age at onset of Alzheimer's disease, and illness duration. Gender, education, and family history of dementia or psychiatric illness showed weak or inconsistent relationships with psychosis. CONCLUSIONS: Psychotic symptoms are common and persistent in patients with Alzheimer's disease. Improved methods have advanced the understanding of psychosis in Alzheimer's disease, although continued research, particularly longitudinal studies, may unveil biological and clinical associations that will inform treatments for these problematic psychological disturbances. PMID- 16263839 TI - Secondary mania in older adults. PMID- 16263840 TI - George Engel, M.D. (1913-1999). PMID- 16263841 TI - Presidential address. PMID- 16263842 TI - Response to the presidential address: advocacy for our patients and our profession. PMID- 16263844 TI - Functional neuroanatomy of successful paired associate learning in Alzheimer's disease. AB - OBJECTIVE: The purpose of the study was to develop a strategy for functional imaging of neurodegenerative disorders that overcomes confounds associated with differential performance between patient and comparison groups. METHOD: Functional magnetic resonance imaging was used to examine responses to increasing difficulty of visuospatial paired associate learning in 12 patients with mild probable Alzheimer's disease and 12 age-matched healthy comparison subjects. Performance was matched across groups by only examining successful encoding and retrieval attempts. Adjustment for task difficulty was made on an individual basis so that the patients with Alzheimer's disease and the comparison subjects performed at the same relative levels of difficulty. RESULTS: A network of lateral and medial frontoparietal and occipital regions was engaged in all subjects during successful associative learning. As task difficulty increased, blood-oxygen-level-dependent responses increased linearly in occipitoparietal regions during encoding and retrieval. Differential activations in patients with Alzheimer's disease and comparison subjects were small and were found only when an uncorrected statistical threshold was used. CONCLUSIONS: By controlling for confounds of varying task difficulty and subsequent performance, remarkably similar brain activations were identified during successful paired associate learning in patients with Alzheimer's disease and in healthy comparison subjects. The study methods provide a useful model for further applications of functional imaging involving cognitive activation paradigms in the study of neuropsychiatric disorders. PMID- 16263845 TI - Pharmacological modulation of prefrontal cortical activity during a working memory task in young and older humans: a PET study with physostigmine. AB - OBJECTIVE: Age-associated cholinergic dysfunction may contribute to the cognitive decline observed during aging, including a decline in working memory. The current study was designed to determine how healthy aging influences the neural response to working memory before and during pharmacological potentiation of the cholinergic system. METHOD: In 13 young and 13 older healthy volunteers, regional cerebral blood flow (rCBF) was measured by using [15O]H2O and positron emission tomography across 10 scans that alternated between a working-memory-for-faces task and rest. In all subjects, the first two scans were obtained during intravenous saline infusion. Seven young and eight older subjects then received intravenous infusion of physostigmine, a cholinesterase inhibitor, and the remaining six young and five older subjects continued to receive saline. RESULTS: In the placebo condition, task-specific rCBF increases in prefrontal regions were observed in the right middle and inferior frontal cortices in young subjects and in more anterior and ventral prefrontal regions in older individuals. Physostigmine during the working memory task significantly improved performance in both age groups. The right prefrontal regions that were selectively recruited in each age group during the placebo condition showed significantly lower rCBF during physostigmine administration. CONCLUSIONS: Cholinergic enhancement does not affect structurally defined cortical regions but rather modulates neural activity in functionally defined regions, that is, in task-related prefrontal cortical areas that are selectively and distinctively recruited in young and older subjects. PMID- 16263846 TI - Prevalence and natural course of aging-associated cognitive decline in a population-based sample of young-old subjects. AB - OBJECTIVE: "Mild cognitive impairment" refers to cognitive deficits in older age that exceed age-related cognitive decline but do not fulfill criteria for dementia. Affected subjects are assumed to be at higher risk for the development of dementia, such as Alzheimer's disease. However, little is known about the group of young-old subjects with respect to the prevalence and natural course of cognitive decline. METHOD: Within the population-based Interdisciplinary Longitudinal Study on Adult Development and Aging, neuropsychological functioning was assessed in 500 community-dwelling young-old subjects of two German urban regions who were born during 1930-1932. The participants were carefully screened for physical and mental health and reexamined 4 years later. The concept of "aging-associated cognitive decline" was applied. RESULTS: At baseline, 13.4% of the subjects fulfilled criteria for aging-associated cognitive decline. Four years later, the prevalence rates for rose to 23.6%; 52.3% of the subjects initially classified as having aging-associated cognitive decline retained the diagnosis at follow-up. Although subjects with aging-associated cognitive decline showed a reduced performance in all neuropsychological domains addressed, a significant decline was confined to delayed verbal memory test performance during the 4-year follow-up period in relation to comparison subjects. Aging-associated cognitive decline did not predict conversion to dementia during the follow-up interval. CONCLUSIONS: In young-old community-dwelling individuals, aging associated cognitive decline is a frequent condition with a high temporal stability. During a 4-year follow-up, subjects with aging-associated cognitive decline deteriorated specifically in measures of episodic memory, underscoring the value of the respective deficits in characterizing "mild cognitive impairment." PMID- 16263847 TI - The pattern of cognitive performance in CADASIL: a monogenic condition leading to subcortical ischemic vascular dementia. AB - OBJECTIVE: Subcortical ischemic vascular lesions, which are closely related to small vessel disease, are a common substrate of cognitive impairment and dementia. Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is a monogenic variant of small vessel disease resulting from mutations in NOTCH3. Mutation carriers almost invariably develop cognitive deficits and eventually dementia. The current study describes the profile of cognitive abnormalities in CADASIL subjects. METHOD: A cross-sectional study of 65 mutation carriers (mean age=47.3 years, SD=10.5) and 30 matched comparison subjects (mean age=47.2 years, SD=14.0) was conducted. Participants underwent a series of assessments that included ratings of global cognition, the cognitive portion of the Vascular Dementia Assessment Scale, and specific tests of executive function and attention with measures of processing speed and error monitoring. RESULTS: CADASIL subjects had pronounced impairments of the timed measures (Stroop II and III, Trail Making Test, symbol digit, digit cancellation). Measures of error monitoring (Stroop III, Trail Making Test, symbol digit, maze task) were also significantly affected but to a lesser extent. Prominent deficits further included verbal fluency and ideational praxis. Recall, orientation, and receptive language skills were largely preserved. Subgroup analyses indicated a similar profile in subjects with early and advanced impairment of global cognitive performance. CONCLUSIONS: The findings highlight processing speed as the most substantial area of cognitive impairment in CADASIL subjects, with less pronounced yet significant deficits in other aspects of executive performance and attention. This profile of cognitive impairment is present at an early stage and enables the construction of targeted test batteries for clinical trials. It is hypothesized that the profile of dysfunction described here represents the core of the cognitive syndrome associated with small vessel disease and subcortical ischemic vascular lesions. PMID- 16263848 TI - The construct of minor and major depression in Alzheimer's disease. AB - OBJECTIVE: This study examined the frequency of major and minor depression in Alzheimer's disease and determined whether these types of depression have a different functional and psychopathological impact and whether there is a change in the prevalence of major and minor depression throughout the stages of Alzheimer's disease. METHOD: A consecutive series of 670 patients with probable Alzheimer's disease were assessed with the Structured Clinical Interview for DSM IV; specific instruments to rate the presence and severity of depression, anxiety, apathy, irritability, delusions, pathological affective crying, performance of activities of daily living, and social functioning; and a standardized neuropsychological evaluation. Diagnoses of major and minor depression were generated from DSM-IV criteria. RESULTS: Twenty-six percent of the patients had major depression, 26% had minor depression, and 48% were not depressed. Major depression was significantly associated with sad mood in all three stages of the illness, although this association dropped significantly for minor depression in severe Alzheimer's disease. Both major and minor depression were significantly associated with more severe psychopathology, functional impairments, and social dysfunction. Depressive symptoms that most strongly discriminated between Alzheimer's disease patients with and without sad mood were guilty ideation, suicidal ideation, loss of energy, insomnia, weight loss, psychomotor retardation/agitation, poor concentration, and loss of interest. CONCLUSIONS: Our study demonstrates that DSM-IV criteria for major and minor depression identify clinically relevant syndromes of depression in Alzheimer's disease, mild levels of depression can produce significant functional impairment, and the severity of psychopathological and neurological impairments increases with increasing severity of depression. PMID- 16263849 TI - The capacity to vote of persons with Alzheimer's disease. AB - OBJECTIVE: The right to vote can be abrogated when persons become incompetent to cast a ballot. This applies particularly to people with Alzheimer's disease, who at some point will lose capacity. A 2001 federal court decision offered the first clear criteria ("Doe voting capacity standard") for determining voting competence, focused on understanding the nature and effect of voting and on the ability to choose. This article explores how persons with Alzheimer's disease perform on these criteria. METHOD: The Doe standard was operationalized in a brief questionnaire, along with measures of appreciation and reasoning about voting choices. Performance was assessed in 33 patients attending an Alzheimer's disease clinic and was related to dementia severity and demographic characteristics. RESULTS: The interview questions were scored with high reliability. Performance on the Doe questions, along with appreciation and reasoning, correlated strongly with Mini-Mental State Examination (MMSE) scores. Patients with very mild to mild Alzheimer's disease generally retained adequate ability to vote, and most persons with severe Alzheimer's disease did not. Performance was highly variable among persons with moderate Alzheimer's disease. The desire to vote was a poor predictor of voting capacity. CONCLUSIONS: The capacity to vote, as embodied in the Doe voting capacity standard, can be measured simply and reliably. Structured assessment is particularly likely to be useful for people with moderate Alzheimer's disease, whose performance cannot be predicted from MMSE scores alone. This approach can ensure retention of voting rights by capable persons and exclusion of clearly impaired persons from the voting booth. PMID- 16263850 TI - Genotype-phenotype studies in bipolar disorder showing association between the DAOA/G30 locus and persecutory delusions: a first step toward a molecular genetic classification of psychiatric phenotypes. AB - OBJECTIVE: The authors previously reported an association between the D-amino acid oxidase activator (DAOA)/G30 locus and both schizophrenia and bipolar affective disorder. Given the presumed role of DAOA/G30 in the neurochemistry of psychosis and its localization in a schizophrenia and bipolar affective disorder linkage region (13q34), it was hypothesized that the bipolar affective disorder finding would be mainly due to an association with psychotic features. METHOD: The marker/haplotype associations obtained in a subset of 173 bipolar affective disorder patients with psychotic features were similar to those in the overall patient group, suggesting that stratification on the basis of psychotic features in general might be too crude a procedure. The authors therefore tested whether confining caseness to specific psychotic features would improve detection of genotype-phenotype correlations. RESULTS: In a logistic regression, "persecutory delusions" were found to be the only significant explanatory variable for the DAOA/G30 risk genotype among 21 OPCRIT symptoms of psychosis. The authors therefore tested for association between DAOA/G30 and bipolar affective disorder in the 90 cases with a history of persecutory delusions. Whereas this subset showed strong association (odds ratio=1.83 for the best marker), the remaining larger sample of 165 patients with no such history did not differ from comparison subjects, suggesting that the association between DAOA/G30 and bipolar affective disorder is due to persecutory delusions. This was confirmed in an independent study of 294 bipolar affective disorder patients and 311 comparison subjects from Poland, in which an association between bipolar affective disorder and DAOA/G30 was only seen when case definition was restricted to cases with persecutory delusions. CONCLUSIONS: These data suggest that bipolar affective disorder with persecutory delusions constitutes a distinct subgroup of bipolar affective disorder that overlaps with schizophrenia. PMID- 16263851 TI - Reduced NAA levels in the dorsolateral prefrontal cortex of young bipolar patients. AB - OBJECTIVE: Converging evidence implicates prefrontal circuits in the pathophysiology of bipolar disorder. Proton spectroscopy studies performed in adult bipolar patients assessing prefrontal regions have suggested decreased levels of N-acetylaspartate (NAA), a putative marker of neuronal integrity. In order to examine whether such abnormalities would also be found in younger patients, a 1H spectroscopy study was conducted that focused on the dorsolateral prefrontal cortex of children and adolescents with bipolar disorder. METHOD: The authors examined the levels of NAA, creatine plus phosphocreatine, and choline containing molecules in the left dorsolateral prefrontal cortex of 14 bipolar disorder patients (mean age=15.5 years, SD=3, eight female) and 18 healthy comparison subjects (mean age=17.3, SD=3.7, seven female) using short echo time, single-voxel in vivo 1H spectroscopy. Absolute metabolite levels were determined using the water signal as an internal reference. RESULTS: Bipolar patients presented significantly lower NAA levels and a significant inverse correlation between choline-containing molecules and number of previous affective episodes. No differences were found for other metabolites. CONCLUSIONS: These findings suggest that young bipolar patients have decreased NAA levels in the dorsolateral prefrontal cortex, similar to what was previously reported in adult patients. Such changes may reflect an underdevelopment of dendritic arborizations and synaptic connections. These neuronal abnormalities in the dorsolateral prefrontal cortex of bipolar disorder youth are unlikely to represent long-term degenerative processes, at least in the subgroup of patients where the illness had relatively early onset. PMID- 16263852 TI - Risk factors for suicide completion in major depression: a case-control study of impulsive and aggressive behaviors in men. AB - OBJECTIVE: Major depression is a major risk factor for suicide. However, not all individuals with major depression commit suicide. Impulsive and aggressive behaviors have been proposed as risk factors for suicide, but it remains unclear whether their effect on the risk of suicide is at least partly explained by axis I disorders commonly associated with suicide, such as major depression. With a case-control design, a comparison of the level of impulsive and aggressive behaviors and the prevalence of associated psychopathology was carried out with control for the presence of primary psychopathology. METHOD: One hundred and four male suicide completers who died during an episode of major depression and 74 living depressed male comparison subjects were investigated with proxy-based interviews by using structured diagnostic instruments and personality trait assessments. RESULTS: The authors found that current (6-month prevalence) alcohol abuse/dependence, current drug abuse/dependence, and cluster B personality disorders increased the risk of suicide in individuals with major depression. Also, higher levels of impulsivity and aggression were associated with suicide. An analysis by age showed that these risk factors were more specific to younger suicide victims (ages 18-40). A multivariate analysis indicated that current alcohol abuse/dependence and cluster B personality disorder were two independent predictors of suicide. CONCLUSIONS: Impulsive-aggressive personality disorders and alcohol abuse/dependence were two independent predictors of suicide in major depression, and impulsive and aggressive behaviors seem to underlie these risk factors. A developmental hypothesis of suicidal behavior, with impulsive and aggressive behaviors as the starting point, is discussed. PMID- 16263853 TI - Association of resolution of major depression with increased natural killer cell activity among HIV-seropositive women. AB - OBJECTIVE: Depression is a potential risk factor for morbidity and mortality among patients with numerous medical conditions, including HIV disease, and it is also associated with decrements in immune function, such as natural killer (NK) cell activity. This study examined whether improvements in the diagnostic status of major depression are related to increases in NK cell activity among HIV seropositive women. METHOD: HIV-seropositive women were recruited as part of a longitudinal cohort study and underwent comprehensive medical and psychiatric evaluations during a 2-year period. Fifty-seven women had complete NK cell activity and depression data measured at two time points and were examined for associations between changes in depression status and alterations in NK cell activity over time. RESULTS: Among the 57 HIV-seropositive women, improvements in the diagnostic status of depression and decreases in scores on the 17-item Hamilton Depression Rating Scale were significantly associated with increases in NK cell activity over time, as measured in lytic units. Eleven women (19.3%) had a major depression diagnosis that resolved over time, and this group also had a significant increase in cell activity measured in lytic units during this period. CONCLUSIONS: This study suggests that depression may impair certain aspects of innate cellular immunity relevant to delaying the progression of HIV disease and that these alterations are reversible with the resolution of a depressive episode. These findings support an examination of NK cell activity in assessments of the relationship between depression and morbidity and mortality in HIV disease. PMID- 16263854 TI - Treatment matching in the posthospital care of depressed patients. AB - OBJECTIVE: This study assessed the efficacy of 1) matching patients to treatments and 2) adding additional family therapy or cognitive therapy in a group of recently discharged patients with major depression. METHOD: Patients with major depression were recruited during a psychiatric hospitalization. After discharge, they were randomly assigned to one of four treatment conditions that were either "matched" or "mismatched" to their pattern of cognitive distortion and family impairment. The four treatment conditions were 1) pharmacotherapy alone; 2) combined pharmacotherapy and cognitive therapy; 3) combined pharmacotherapy and family therapy; and 4) combined pharmacotherapy, cognitive therapy, and family therapy. Randomly assigned treatment continued for 24 weeks on an outpatient basis. RESULTS: Among patients with at least moderate depressive symptoms at hospital discharge, low rates of remission (16%) and improvement (29%) were obtained. Matched treatment led to a significantly greater proportion of patients who improved and greater reductions over time in interviewer-rated depressive symptoms than mismatched treatment. However, matched treatment did not produce greater change in self-reported depression or interviewer-rated suicidal ideation. Treatment that included a family therapy component also led to a greater proportion of patients who improved and to significant reductions in interviewer-rated depression and suicidal ideation than treatment without family therapy. CONCLUSIONS: These results suggest that 1) current treatments are not very efficacious in the aftercare of hospitalized depressed patients, 2) treatment matching moderately improves outcome for patients who are symptomatic at hospital discharge, and 3) inclusion of family therapy improves the outcome of posthospital care for depressed patients. PMID- 16263855 TI - Depressive symptoms and 24-hour urinary norepinephrine excretion levels in patients with coronary disease: findings from the Heart and Soul Study. AB - OBJECTIVE: Depressive symptoms are associated with an increased risk of cardiac events in patients with heart disease. Elevated catecholamine levels may contribute to this association, but whether depressive symptoms are associated with catecholamine levels in patients with heart disease is unknown. METHOD: The authors examined the association between depressive symptoms (defined by a Patient Health Questionnaire score > or =10) and 24-hour urinary norepinephrine, epinephrine, and dopamine excretion levels in 598 subjects with coronary disease. RESULTS: A total of 106 participants (18%) had depressive symptoms. Participants with depressive symptoms had greater mean norepinephrine excretion levels than those without depressive symptoms (65 microg/day versus 59 mug/day, with adjustment for age, sex, body mass index, smoking, urinary creatinine levels, comorbid illnesses, medication use, and cardiac function). In logistic regression analyses, participants with depressive symptoms were more likely than those without depressive symptoms to have norepinephrine excretion levels in the highest quartile and above the normal range. Depressive symptoms were not associated with dopamine or epinephrine excretion levels. CONCLUSIONS: In patients with coronary disease, depressive symptoms are associated with elevated norepinephrine excretion levels. Future longitudinal studies are needed to determine whether elevations in norepinephrine contribute to adverse cardiac outcomes in patients with depressive symptoms. PMID- 16263857 TI - A 20-month, double-blind, maintenance trial of lithium versus divalproex in rapid cycling bipolar disorder. AB - OBJECTIVE: The authors tested the hypothesis that divalproex would be more effective than lithium in the long-term management of patients with recently stabilized rapid-cycling bipolar disorder. METHOD: A 20-month, double-blind, parallel-group comparison was carried out in recently hypomanic/manic patients who had experienced a persistent bimodal response to combined treatment with lithium and divalproex. Sixty patients were randomly assigned to lithium or divalproex monotherapy in a balanced design after stratification for illness type (bipolar I versus bipolar II disorder). RESULTS: Of the 254 patients enrolled in the open-label acute stabilization phase, 76% discontinued the study prematurely (poor adherence: 28%; nonresponse: 26% [of whom 74% remained depressed and 26% remained in a hypomanic/manic/mixed episode], intolerable side effects: 19%). Of the 60 patients (24%) randomly assigned to double-blind maintenance monotherapy, 53% relapsed (59% into depression and 41% into a hypomanic/manic/mixed episode), 22% completed the study, 10% had intolerable side effects, and 10% were poorly adherent. The rates of relapse into any mood episode for those given lithium versus divalproex were 56% and 50%, respectively; the rates were 34% and 29% for a depressive relapse and 19% and 22% for a hypomania/mania relapse. There were no significant differences in time to relapse. The proportion discontinuing prematurely because of side effects was 16% for lithium and 4% for divalproex. CONCLUSIONS: The hypothesis that divalproex is more effective than lithium in the long-term management of rapid-cycling bipolar disorder is not supported by these data. Preliminary data suggest highly recurrent refractory depression may be the hallmark of rapid-cycling bipolar disorder. PMID- 16263856 TI - Bipolar depression in a low-income primary care clinic. AB - OBJECTIVE: This study estimated the proportion of patients attending an urban general medical practice with current major depression and a history of bipolar disorder and compared the history, presentation, and treatment of patients with unipolar and bipolar depression. METHOD: A group of 1,143 patients was assessed with measures of past and current mental health and treatment. Patients were partitioned into bipolar and unipolar groups based on a predefined cutoff on the Mood Disorder Questionnaire. The groups were compared on sociodemographic characteristics, depressive symptoms, comorbid mental disorders, and mental health treatment. RESULTS: Approximately one-quarter of the patients with major depression had lifetime bipolar depression. Patients with unipolar and bipolar depression did not significantly differ on background or health characteristics. Patients with bipolar depression were significantly more likely to report hallucinations, current suicidal ideation, and low self-esteem than patients with unipolar depression but less likely to report disturbed appetite. Patients with bipolar depression were significantly more likely to have an alcohol use disorder and to report inpatient psychiatric care and antipsychotic treatment during the past month than patients with unipolar depression. Nearly one-half of the patients with bipolar depression had taken an antidepressant in the last month, but most were not also being treated with an antipsychotic or mood stabilizer. CONCLUSIONS: Bipolar depression is common in urban general medicine practice. When patients took antidepressants, they seldom received concurrent antimanic medications. Because of the risks of treating bipolar disorder with antidepressant monotherapy, physicians should assess their depressed patients for mania before prescribing antidepressants. PMID- 16263858 TI - Lithium placental passage and obstetrical outcome: implications for clinical management during late pregnancy. AB - OBJECTIVE: Lithium has been used during pregnancy for more than four decades, but quantification of fetal lithium exposure and clinical correlations of such exposure are limited. The study objectives were to 1) quantify the rate of lithium placental passage, 2) assess any association between plasma concentration of lithium at delivery and adverse perinatal events, and 3) determine whether lithium concentrations can be reduced by briefly suspending therapy proximate to delivery. METHOD: Maternal blood and umbilical cord blood were obtained at delivery for assay of lithium concentrations, and obstetrical outcome data were collected prospectively for 10 participants. These data were combined with results from MEDLINE and PsycINFO searches that identified 32 cases in which maternal lithium was administered throughout delivery. Statistical analysis of the pooled data was conducted. RESULTS: The ratio of lithium concentrations in umbilical cord blood to maternal blood (mean=1.05, SD=0.13) was uniform across a wide range of maternal concentrations (0.2-2.6 meq/liter). Significantly lower Apgar scores, longer hospital stays, and higher rates of CNS and neuromuscular complications were observed in infants with higher lithium concentrations (>0.64 meq/liter) at delivery. Withholding lithium therapy for 24-48 hours before delivery resulted in a 0.28 meq/liter reduction in maternal lithium concentration. CONCLUSIONS: Lithium completely equilibrates across the placenta. Higher lithium concentrations at delivery are associated with more perinatal complications, and lithium concentrations can be reduced by brief suspension of therapy proximate to delivery. Treatment guidelines are proposed to improve neonatal well-being when lithium use is indicated in late pregnancy. PMID- 16263859 TI - Mood-congruent bias in affective go/no-go performance of unmedicated patients with major depressive disorder. AB - OBJECTIVE: Neuropsychological studies of major depressive disorder have described attentional biases for affectively laden stimuli, but these reports were based on measures obtained from medicated subjects. This study investigated performance of unmedicated depressed patients on the Affective Go/No-Go Task. METHOD: Twenty depressed patients and 20 healthy comparison subjects, matched for age, gender, and IQ, performed the Affective Go/No-Go Task as well as tests of attention and memory for nonaffective stimuli. RESULTS: Depressed patients did not differ from healthy subjects on memory task performance, but they made more omission errors on the attention task. On the Affective Go/No-Go Task, depressed patients made more omission errors during happy than sad word blocks and required more time to respond to happy than to sad words. In contrast, healthy subjects required more time to respond to sad than to happy words. CONCLUSIONS: Unmedicated depressed patients do not show a pattern of generalized cognitive impairment but, rather, specifically display an attentional deficit and a mood-congruent bias toward salient stimuli. PMID- 16263860 TI - Emergent suicidality in a clinical psychotherapy trial for adolescent depression. AB - OBJECTIVE: The authors' goal was to examine the incidence and predictors of emergent suicidality that occurred during a clinical trial of psychotherapy for adolescent depression. METHOD: The rates and predictors of emergent suicidality in 88 medication-free depressed adolescent outpatients who reported no current suicidality during an intake interview were assessed over 12 to 16 weeks of psychotherapy treatment. RESULTS: The incidence of emergent suicidality was 12.5% (11 of 88 subjects). Self-reported suicidal thoughts at intake were a significant predictor of emergent suicidality, even when suicidality was denied at intake interview. CONCLUSIONS: Emergent suicidality is a common occurrence in psychosocial treatment of adolescent depression, with rates similar to those reported recently in antidepressant trials. To evaluate accurately the role of treatment in emergent suicidality, it is important to assess self-reported suicidality at intake and to balance treatment groups on this key predictor of emergent suicidality. PMID- 16263861 TI - Complicated grief, depression, and anxiety as distinct postloss syndromes: a confirmatory factor analysis study. AB - OBJECTIVE: The authors used confirmatory factor analysis to replicate earlier findings that complicated grief, depression, and anxiety are distinct syndromes. METHOD: Data were derived from 1,321 bereaved individuals. Complicated grief was measured with the Inventory of Traumatic Grief. Depression and anxiety were measured with the SCL-90. RESULTS: A model in which symptoms of complicated grief, depression, and anxiety loaded on separate factors was superior to a one factor model, revealed good model fit, and was invariant across subgroups. CONCLUSIONS: Previous findings of a distinction among complicated grief, depression, and anxiety were confirmed. PMID- 16263862 TI - The broken heart: suicidal ideation in bereavement. AB - OBJECTIVE: This report examined suicidal behavior during bereavement. METHOD: Suicidal ideation was examined in a group of 60 recently bereaved widows and widowers compared to 60 individually matched married comparison subjects. RESULTS: Suicidal ideation was higher among widowed people than married people and was most excessive for widows. The effect disappeared when there was control for emotional loneliness. Social support did not buffer bereaved individuals against suicidal ideation but reduced suicidal ideation among the married and bereaved alike. CONCLUSIONS: Bereaved persons are at excess risk of suicidal ideation compared to nonbereaved people. Heightened suicidal ideation in bereavement is associated with extreme emotional loneliness and severe depressive symptoms. PMID- 16263863 TI - Suicide attempters' reaction to survival as a risk factor for eventual suicide. AB - OBJECTIVE: The authors' goal was to evaluate whether suicide attempters' reaction to surviving their attempt predicted eventual suicide. METHOD: Three hundred ninety-three suicide attempters were categorized on the basis of their reaction to having survived their attempt (i.e., glad to be alive, ambivalent, wished they were dead) and were followed for 5 to 10 years to determine whether they completed suicide. RESULTS: A survival analysis found that subjects who said that they wished they had died after a suicide attempt were 2.5 times more likely to commit suicide eventually than those who were glad they survived and those who were ambivalent about the attempt. CONCLUSIONS: Suicide attempters' reaction to surviving is an important clinical variable that is easily assessed in evaluations that occur following a suicide attempt. PMID- 16263864 TI - Confirmation of association between autism and the mitochondrial aspartate/glutamate carrier SLC25A12 gene on chromosome 2q31. AB - OBJECTIVE: Autism is a neurodevelopmental disorder with childhood onset and a known major genetic component. A recent study identified a highly significant association between autism and a two-single-nucleotide-polymorphism haplotype in the SLC25A12 gene, with a homozygote genotype relative risk between 2.4 and 4.8. The authors' goal was to investigate this association with autism in Irish affected child-parent trios because replication in an independent sample is essential in the validation of such potentially important findings. METHOD: Markers rs2056202 and rs2292813 were genotyped in a total of 158 trios (442 individuals). The Transmission Disequilibrium Test was used to examine these markers for association with autism. RESULTS: In agreement with the recent study, the authors found significant association between autism and the C alleles of both rs2056202 and rs2292813 as well as the two-marker haplotype. CONCLUSIONS: These findings provide replication of the association between autism and SLC25A12. PMID- 16263865 TI - Impulse control disorders in adult psychiatric inpatients. AB - OBJECTIVE: The authors' goal was to examine the prevalence of impulse control disorders in psychiatric inpatients. METHOD: They used the Minnesota Impulsive Disorders Interview, a semistructured clinical interview assessing pathological gambling, trichotillomania, kleptomania, pyromania, intermittent explosive disorder, compulsive buying, and compulsive sexual behavior, to screen 204 consecutively admitted psychiatric inpatients. One hundred twelve of the inpatients were women (54.9%), and the mean age of the 204 inpatients was 40.5 years (SD=13.2, range=18-83). Patients whose screen was positive for an impulse control disorder were evaluated with structured clinical interviews. RESULTS: Sixty-three patients (30.9%) were diagnosed with at least one current impulse control disorder. The most common impulse control disorders were compulsive buying (N=19 [9.3%]), kleptomania (N=16 [7.8%]), and pathological gambling (N=14 [6.9%]). Patients with and without co-occurring impulse control disorders did not differ significantly from each other on demographic measures or number or type of psychiatric diagnoses other than impulse control disorders. CONCLUSIONS: Impulse control disorders appear common among psychiatric inpatients. Additional, larger studies are needed to examine the prevalence of impulse control disorders in the general population and specific psychiatric groups. PMID- 16263866 TI - Clinical presentations and treatment outcomes of peacekeeper veterans with PTSD: preliminary findings. AB - OBJECTIVE: Despite evidence of potential psychiatric sequelae following peacekeeping operations, no data have appeared on treatment outcome for this population. This study examined intake and treatment outcome data for a group of peacekeepers with posttraumatic stress disorder (PTSD). METHOD: Participants were 63 Australian Vietnam veterans and 66 Australian peacekeepers attending specialized PTSD treatment units. Measures of PTSD, depression, anxiety, alcohol use, and anger were obtained at intake and 3-month follow-up. RESULTS: PTSD scores were more severe for peacekeepers than Vietnam veterans at intake, primarily in reexperiencing symptoms. In terms of comorbidity, only anger was higher among peacekeepers. No differences were apparent in treatment outcome. Initial anger predicted change in PTSD severity for peacekeepers. CONCLUSIONS: The finding of differences between peacekeepers and Vietnam veterans in anger and reexperiencing symptoms, in addition to the attenuating role of anger on treatment outcome, suggests that modification to standard PTSD treatment models may be warranted for peacekeepers. PMID- 16263867 TI - Memantine for treatment-resistant OCD. PMID- 16263868 TI - Tardive dystonia associated with ziprasidone. PMID- 16263870 TI - Deep brain stimulation for OCD and major depression. PMID- 16263869 TI - Therapeutic brain stimulation and cortical excitability in depressed patients. PMID- 16263871 TI - Understanding the heterogeneity of OCD. PMID- 16263872 TI - Heart transplantation in a schizophrenia patient. PMID- 16263873 TI - Conclusions inconsistent with results with citalopram. PMID- 16263874 TI - Insight and aggression in schizophrenia. PMID- 16263875 TI - Conclusions inconsistent with results with amphetamines and divalproex. PMID- 16263876 TI - The Multiple Sleep Latency Test in the diagnosis of narcolepsy. PMID- 16263877 TI - Psychiatric effects of ephedra: addiction. PMID- 16263878 TI - Suicide among physicians. PMID- 16263880 TI - Summaries for patients. Relapse and treatment resistance in patients with small vessel vasculitis. PMID- 16263881 TI - Summaries for patients. Does curing childhood cancer lead to a normal life? PMID- 16263882 TI - Summaries for patients. Hepatitis C virus transmission to organ and tissue recipients. PMID- 16263883 TI - Summaries for patients. The effectiveness of cardiac rehabilitation programs with and without exercise components. PMID- 16263884 TI - Predictors of relapse and treatment resistance in antineutrophil cytoplasmic antibody-associated small-vessel vasculitis. AB - BACKGROUND: Predictors of treatment resistance and relapse have not been well described in antineutrophil cytoplasmic antibody (ANCA)-associated small-vessel vasculitis. OBJECTIVE: To identify clinical, pathologic, and serologic predictors of treatment resistance and relapse in a community-based cohort of patients with ANCA-associated vasculitis. DESIGN: Cohort of patients identified at or near the time of biopsy diagnosis and followed as clinically indicated. SETTING: The Glomerular Disease Collaborative Network. PATIENTS: 350 patients who received a new diagnosis of ANCA-associated vasculitis between 1985 and 2003 and were followed for a median of 49 months. MEASUREMENTS: Patients were categorized according to whether they had antiproteinase-3 (anti-PR3) antibodies or antimyeloperoxidase (anti-MPO) antibodies. Organ involvement was determined by biopsy or by well-defined clinical criteria. Treatment resistance was defined as progressive decline in kidney function with active urine sediment or the persistence or appearance of extrarenal manifestations. Relapse was defined as the time to the resurgence of vasculitic symptoms. RESULTS: Treatment resistance affected 23% of 334 treated patients and was associated with female sex, black ethnicity, and presentation with severe kidney disease (odds ratio per serum creatinine elevation of 100 micromol/L [1.13 mg/dL], 1.28 [95% CI, 1.16 to 1.39]). The following factors were associated with relapse in 258 (77%) patients who attained remission: seropositivity for anti-PR3 antibodies (hazard ratio, 1.87 [CI, 1.11 to 3.14]) and disease of the lung (hazard ratio, 1.71 [CI, 1.04 to 2.81]) or upper respiratory tract (hazard ratio, 1.73 [CI, 1.04 to 2.88]). Relapses occurred in 26% of patients with no risk factors versus 73% of patients with all 3 risk factors (hazard ratio, 3.7 [CI, 1.4 to 9.7]). Among 143 patients attaining remission who subsequently stopped all immunosuppressant therapy, relapse rates were similar for those who had received cyclophosphamide therapy for 6 months or less (34%) compared with those treated for a longer duration (35%), even after adjusting for risk factors for relapse (hazard ratio, 1.41 [CI, 0.80 to 2.50]). LIMITATIONS: The cohort mostly included patients with biopsy proven kidney disease. Patients were not followed with uniform treatment protocols, and only limited information about their clinical course before diagnosis was available. CONCLUSIONS: Female or black patients, or those with severe kidney disease, may be resistant to initial treatment more often than other patients with ANCA-associated small-vessel vasculitis. Increased risk for relapse appears to be related to the presence of lung or upper airway disease and anti-PR3 antibody seropositivity. PMID- 16263885 TI - Antineutrophil cytoplasmic antibodies and the Churg-Strauss syndrome. AB - BACKGROUND: Since testing for antineutrophil cytoplasmic antibodies (ANCA) became available for routine evaluation, no large homogeneous cohort of patients with the Churg-Strauss syndrome has been studied. OBJECTIVE: To define the clinical and biological characteristics of newly diagnosed Churg-Strauss syndrome, according to the presence or absence of ANCA. DESIGN: Cross-sectional analysis of manifestations of participants who were enrolled in treatment trials between December 1995 and December 2002. SETTING: Multicenter study in 63 clinical centers in France, Belgium, Latvia, and the United Kingdom, coordinated by the French Vasculitis Study Group. PARTICIPANTS: 112 patients with Churg-Strauss syndrome that was recently diagnosed on the basis of current classifications. MEASUREMENTS: The authors compared principal demographic, clinical, and laboratory features according to ANCA status at diagnosis. RESULTS: The authors detected ANCA in 43 (38%) patients. Positive ANCA status at diagnosis was associated with renal involvement, peripheral neuropathy, and biopsy-proven vasculitis, whereas negative ANCA status was associated with heart disease and fever. LIMITATIONS: The authors assessed ANCA by immunofluorescence, but they did not assess ANCA centrally or systematically retest if ANCA was undetected at diagnosis. CONCLUSIONS: Phenotypically, ANCA-positive and ANCA-negative Churg Strauss syndrome might differ. The association of ANCA positivity with clinical symptoms that indicate inflammation and necrosis of small vessels might characterize a predominantly vasculitic pattern of the Churg-Strauss syndrome. PMID- 16263886 TI - Limitations on physical performance and daily activities among long-term survivors of childhood cancer. AB - BACKGROUND: Survivors of childhood cancer may experience important disease- and treatment-related late effects, including functional limitations. OBJECTIVE: This study evaluated performance limitations and restricted abilities to participate in personal care, to engage in routine activities like shopping or housework, and to attend work or school (participation restrictions) in a cohort of survivors of childhood cancer. SETTING: Epidemiologic survey and 26 institutions that treat childhood cancer. PATIENTS: Participants included 11 481 persons who were treated for primary brain cancer, leukemia, Hodgkin disease, non-Hodgkin lymphoma, kidney tumor, neuroblastoma, soft-tissue sarcoma, or malignant bone tumor before the age of 21 years and who survived at least 5 years after diagnosis. The comparison group included 3839 siblings of survivors of childhood cancer. MEASUREMENT: Medical data were abstracted, and participants or parents (if the participants were <18 years of age at survey completion) completed a 24-page questionnaire. RESULTS: Compared with siblings, survivors were more likely to report performance limitations (risk ratio, 1.8 [95% CI, 1.7 to 2.0]) and to report restricted participation in personal care skills (risk ratio, 4.7 [CI, 3.0 to 7.2]), routine activities (risk ratio, 4.7 [CI, 3.6 to 6.2]), and the ability to attend work or school (risk ratio, 5.9 [CI, 4.5 to 7.6]). Survivors of brain (26.6%) and bone (36.9%) cancer were most likely to report performance limitations, restricted ability to do routine activities (20.9% and 8.5%, respectively), and restricted ability to attend work or school (20.0% and 11.2%, respectively). Survivors of brain cancer were also most likely to report restricted abilities to perform personal care (10.5%). LIMITATIONS: There was the potential for participants to be healthier or more physically capable than nonparticipants or for persons to be more motivated to participate in this study if they had functional deficits. In addition, the nature of the questionnaire did not allow specific physical limitations to be measured. CONCLUSION: Long-term survivors of childhood cancer are at increased risk for functional limitations in physical performance and in participation in activities needed for daily living. PMID- 16263887 TI - Transmission of hepatitis C virus to several organ and tissue recipients from an antibody-negative donor. AB - BACKGROUND: Although hepatitis C virus (HCV) transmission through tissue transplantation has been rarely reported, a donor with undetected viremia may infect several recipients. A patient developed acute hepatitis C shortly after tissue transplantation. Ninety-one tissues or organs had been recovered from the donor. OBJECTIVE: To determine whether the donor was the source of infection and the extent of transmission to other organ and tissue recipients. DESIGN: Descriptive epidemiologic study; serum testing for HCV infection. SETTING: Recipients were located in 16 states and 2 other countries. PARTICIPANTS: Donor and graft recipients. MEASUREMENTS: Hepatitis C virus infection was defined as the presence of anti-HCV or HCV RNA. The authors determined the genetic relatedness of viral isolates from the donor and recipients by genotype comparison and quasi-species analysis. RESULTS: The donor was anti-HCV-negative but was HCV RNA-positive (genotype 1a). Forty persons received transplants during 22 months. Five persons were HCV-infected before transplantation or had a genotype other than 1a, and 5 persons had no post-transplantation serum specimens available. Of the remaining 30 recipients, HCV infection occurred in 8 recipients: 3 of 3 organ recipients, 1 of 2 saphenous vein recipients, 1 of 3 tendon recipients, and 3 of 3 tendon with bone recipients. These 8 recipients had viral isolates genetically related to those of the donor. No cases occurred in recipients of skin (n = 2), cornea (n = 1), or irradiated bone (n = 16). LIMITATIONS: Post-transplantation serum specimens were unavailable for 5 recipients. CONCLUSIONS: An anti-HCV-negative donor was the source of HCV infection for 8 recipients of organs or tissues. Although HCV transmission from anti-HCV-negative donors is probably uncommon, changes in donor screening to include routine testing for HCV RNA merit further consideration to improve the safety of transplantation. PMID- 16263888 TI - The academic hospice. AB - The academic hospice is a recent development in health care. Hospice programs and hospitals evolved from the same historical roots in Greek and Roman medicine. The academic hospital emerged as a place where patient care, education, and research are pursued as inextricable parts of the mission. The unique role of the academic medical center in health care is supported by the government, the medical profession, and the public. This article provides a perspective on the emergence of the academic hospice. Dr. Cicely Saunders, who died on 14 July 2005, founded the first such hospice in London, England, in 1967. The authors show that the philosophy of hospice care has the same historical roots as standard health care and describe those elements that distinguish academic hospice programs from other kinds of hospice programs. Finally, the authors note that demographic and economic challenges in the United States and elsewhere only increase the need for academic hospice programs. PMID- 16263890 TI - Update in endocrinology. PMID- 16263889 TI - Meta-analysis: secondary prevention programs for patients with coronary artery disease. AB - BACKGROUND: Although supervised exercise programs reduce mortality in survivors of myocardial infarction, the effects of other types of cardiac secondary prevention programs are unknown. PURPOSE: To determine the effectiveness of secondary cardiac prevention programs with and without exercise components. DATA SOURCES: The authors searched MEDLINE (1966-2004), the Cochrane Central Register of Controlled Trials, EMBASE, CINAHL, SIGLE, and the Cochrane Effective Practice and Organization of Care Study Registry. They also contacted primary study authors and hand-searched bibliographies provided by the Centers for Medicare & Medicaid Services. STUDY SELECTION: Randomized clinical trials. DATA EXTRACTION: Two reviewers chose studies and extracted data independently; random-effects summary risk ratios were calculated. DATA SYNTHESIS: The authors identified 63 randomized trials (21 295 patients with coronary disease). The summary risk ratio was 0.85 (95% CI, 0.77 to 0.94) for all-cause mortality, but this result differed over time with a risk ratio of 0.97 (CI, 0.82 to 1.14) at 12 months and 0.53 (CI, 0.35 to 0.81) at 24 months. The summary risk ratio was 0.83 (CI, 0.74 to 0.94) for recurrent myocardial infarction over a median follow-up of 12 months. Effects were similar for programs that included risk factor education or counseling with a structured exercise component (risk ratio, 0.88 [CI, 0.74 to 1.04] for mortality and 0.62 [CI, 0.44 to 0.87] for myocardial infarction), for programs that included risk factor education or counseling without an exercise component (risk ratio, 0.87 [CI, 0.76 to 0.99] for mortality and 0.86 [CI, 0.72 to 1.03] for myocardial infarction), and for programs that were solely exercise-based (risk ratio, 0.72 [CI, 0.54 to 0.95] for mortality and 0.76 [CI, 0.57 to 1.01] for myocardial infarction). Most of these programs improved quality of life or functional status, but effect sizes were small. LIMITATIONS: Although these programs may reduce total health care costs, published data on the costs of the programs are inadequate to conclusively comment on their cost-effectiveness. CONCLUSIONS: A wide variety of secondary prevention programs improve health outcomes in patients with coronary disease. PMID- 16263891 TI - Are there different forms of life in the antineutrophil cytoplasmic antibody universe? PMID- 16263892 TI - Lessons from a label maker. PMID- 16263893 TI - Failing the grandparent test. PMID- 16263894 TI - Deactivating implantable cardioverter defibrillators. PMID- 16263895 TI - Deactivating implantable cardioverter defibrillators. PMID- 16263896 TI - Acupuncture for low back pain. PMID- 16263897 TI - Deactivating implantable cardioverter defibrillators. PMID- 16263898 TI - The future of generalism in medicine. PMID- 16263899 TI - Cimetidine and acute intermittent porphyria. PMID- 16263900 TI - The role of monovalent cation transporters in plant responses to salinity. AB - Exposure to high ambient levels of NaCl affects plant water relations and creates ionic stress in the form of the cellular accumulation of Cl- and, in particular, Na+ ions. However, salt stress also impacts heavily on the homeostasis of other ions such as Ca2+, K+, and NO(3)(-) and therefore requires insights into how transport and compartmentation of these nutrients is altered during salinity stress. A genomics approach can greatly help with the identification of genes, and therefore potentially gene products, that are involved in plant salinity. Both the literature and public databases contain the results of many genomics studies and, in this report, those data are collated in the context of cation membrane transport and salinity. The efficacy of genomics approaches in isolation is low due to large inherent variability and the exclusion of gene products that are predominantly regulated post-transcriptionally. In conjunction with complementary approaches, however, transcriptomics can help identify important transcripts and relevant associations between physiological processes. This analysis identified (i) vascular K+ circulation, (ii) root shoot translocation of Ca2+, and (iii) transition metal homeostasis as potentially important aspects of the plant response to salt stress. PMID- 16263901 TI - Nitric oxide is involved in abscisic acid-induced antioxidant activities in Stylosanthes guianensis. AB - Previous studies suggest that abscisic acid (ABA) stimulates the activities of antioxidant enzymes under normal and chilling temperature and enhanced chilling resistance in Stylosanthes guianensis. The objective of this study was to test whether nitric oxide (NO) is involved in the ABA-induced activities of the antioxidant enzymes in Stylosanthes guianensis due to its nature as a second messenger in stress responses. Plants were treated with NO donors, ABA, ABA in combination with NO scavengers or the nitric oxide synthase (NOS) inhibitor and their effects on the activity of antioxidant enzymes and NO production were compared. The results showed that ABA increased the activities of superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX). The effect of ABA on antioxidant enzyme activities was suppressed by the NOS inhibitor, N(omega)-nitro-L-arginine (L-NNA), and the NO scavenger, 2-phenyl-4,4,5,5 tetramethylimidazoline-1-oxyl3-oxide (PTIO). NO content increased after 5 h of ABA treatment. The NO-scavenger, PTIO, and the NOS-inhibitor, L-NNA, inhibited the accumulation of NO in ABA-treated Stylosanthes guianensis. NO donor treatment enhanced the activities of SOD, CAT, and APX. The results suggested that NO was involved in the ABA-induced activities of SOD, CAT, and APX in Stylosanthes guianensis. ABA triggered NO production that may lead to the stimulation of antioxidant enzyme activities. PMID- 16263902 TI - Both hypersensitive and non-hypersensitive responses are associated with resistance in Salix viminalis against the gall midge Dasineura marginemtorquens. AB - Hypersensitivity responses (HR) play a major role in plant resistance to pathogens. It is often claimed that HR is also important in plant resistance to insects, although there is little unambiguous documentation. Large genotypic variation in resistance against the gall midge Dasineura marginemtorquens is found in Salix viminalis. Variation in larval performance and induced responses within a full-sib S. viminalis family is reported here; 36 sibling plants were completely resistant (larvae died within 48 h after egg hatch, no gall induction), 11 plants were totally susceptible, 25 plants were variable (living and dead larvae present on the same plant). Resistance was associated with HR, but to different degrees; 21 totally resistant genotypes showed typical HR symptoms (many distinct necrotic spots) whereas the remaining 15 genotypes showed no, or very few, such symptoms. Hydrogen peroxide, used as a marker for HR, was induced in genotypes expressing HR symptoms but not in resistant genotypes without symptoms, or in susceptible genotypes. These data suggest that production of hydrogen peroxide, and accompanying cell death, cannot explain larval mortality in the symptomless reaction. Another, as yet unknown, mechanism of resistance may be present. If so, then it is possible that this unknown mechanism also contributes to resistance in plants displaying HR. The apparent complexity observed in this interaction, with both visible and invisible plant responses associated with resistance against an adapted insect species, may have implications for the study of resistance factors in other plant-insect interactions. PMID- 16263904 TI - Differential expression of genes encoding protein kinase CK2 subunits in the plant cell cycle. AB - Protein kinase CK2 is a ubiquitous Ser/Thr/Tyr kinase essential for cell viability in eukaryotes. It comprises alpha catalytic and beta regulatory subunits, which combine to form the classical tetrameric structure, alpha2beta2. Although CK2 is a component of the network that controls the eukaryotic cell cycle, very little is known about the expression patterns of genes encoding its constituent subunits, especially in plants. A study of the complexity of CK2alpha and CK2beta-encoding genes in BY-2 cells was undertaken in this work, and cloning of the different members of the gene families was performed. The expression of the individual members of each family in relation to cell proliferation was measured by real time RT-PCR. The data obtained provide an accurate understanding of the transcriptional regulation of CK2 in relation to the cell cycle and cell proliferation. PMID- 16263903 TI - OsZIP4, a novel zinc-regulated zinc transporter in rice. AB - Zinc (Zn) is an essential element for the normal growth of plants but information is scarce on the mechanisms whereby Zn is transported in rice (Oryza sativa L.) plants. Four distinct genes, OsZIP4, OsZIP5, OsZIP6, and OsZIP7 that exhibit sequence similarity to the rice ferrous ion transporter, OsIRT1, were isolated. Microarray and northern blot analysis revealed that OsZIP4 was highly expressed under conditions of Zn deficiency in roots and shoots. Real-time-PCR revealed that the OsZIP4 transcripts were more abundant than those of OsZIP1 or OsZIP3 in Zn-deficient roots and shoots. OsZIP4 complemented a Zn-uptake-deficient yeast (Saccharomyces cerevisiae) mutant, Deltazrt1,Deltazrt2, indicating that OsZIP4 is a functional transporter of Zn. OsZIP4-synthetic green fluorescent protein (sGFP) fusion protein was transiently expressed in onion epidermal cells localized to the plasma membrane. In situ hybridization analysis revealed that OsZIP4 in Zn deficient rice was expressed in shoots and roots, especially in phloem cells. Furthermore, OsZIP4 transcripts were detected in the meristem of Zn-deficient roots and shoots. These results suggested that OsZIP4 is a Zn transporter that may be responsible for the translocation of Zn within rice plants. PMID- 16263905 TI - Biochemical and immunohistochemical analysis of pectic polysaccharides in the cell walls of Arabidopsis mutant QUASIMODO 1 suspension-cultured cells: implications for cell adhesion. AB - Mutation in the Arabidopsis thaliana QUASIMODO 1 gene (QUA1), which encodes a putative glycosyltransferase, reduces cell wall pectin content and cell adhesion. Suspension-cultured calli were generated from roots of wild-type (wt) and qua1-1 A. thaliana plants. The altered cell adhesion phenotype of the qua1-1 plant was also found with its suspension-cultured calli. Cell walls of both wt and qua1-1 calli were analysed by chemical, enzymatic and immunohistochemical techniques in order to assess the role of pectic polysaccharides in the mutant phenotype. Compared with the wt, qua1-1 calli cell walls contained more arabinose (23.6 versus 21.6 mol%), rhamnose (3.1 versus 2.7 mol%), and fucose (1.4 versus 1.2 mol%) and less uronic acid (24.2 versus 27.6 mol%), and they were less methyl esterified (DM: 22.9% versus 30.3%). When sequential pectin extraction of calli cell walls was performed, qua1-1 water-soluble and chelator-soluble extracts contained more arabinose and less uronic acid than wt. Water-soluble pectins were less methyl-esterified in qua1-1 than in wt. Chelator-soluble pectins were more acetyl-esterified in qua1-1. Differences in the cell wall chemistry of wt and mutant calli were supported by a reduction in JIM7 labelling (methyl-esterified homogalacturonan) of the whole wall in small cells and particularly by a reduced labelling with 2F4 (calcium-associated homogalacturonan) in the middle lamella at tricellular junctions of large qua1-1 cells. Differences in the oligosaccharide profile obtained after endopolygalacturonase degradation of alkali extracts from qua1-1 and wt calli indicated variations in the structure of covalently bonded homogalacturonan. About 29% more extracellular polymers rich in pectins were recovered from the calli culture medium of qua1-1 compared with wt. These results show that perturbation of QUASIMODO 1-1 gene expression in calli resulted in alterations of homogalacturonan content and cell wall location. The consequences of these structural variations are discussed with regard to plant cell adhesion. PMID- 16263906 TI - A multifunctional bicupin serves as precursor for a chromosomal protein of Pisum sativum seeds. AB - The fact that the psp54 gene codes for p16, a seed chromatin protein of Pisum sativum, has been described previously. In the present paper it is shown that p54, the p16 precursor, also exists as a free polypeptide in pea and that it also yields p38, a second polypeptide from the N-terminal region of p54, which is co localized at a subcellular level with p16. By using antibodies against pea p16 and p38, it was found that these proteins are present in the members of the tribe Viciae examined. Sequence analysis and 3D modelling indicates that p54 proteins belong to the cupin superfamily, and that they are related to sucrose binding proteins and, to a lesser extent, to vicilin-type seed storage proteins. Nevertheless, several distinctive characteristics of psp54 expression have been found: (i) the gene is differentially induced by ABA and several stress situations, in accordance with the presence of putative separate ABA and stress responsive elements in its promoter; (ii) the proteins are present in pods and seed coats, tissues of maternal origin; and (iii) p54 mRNA accumulates in the dry seeds. In view of both the functional properties of p54-derived proteins and the features of the psp54 gene expression, it is concluded that p54 represents a novel class within the cupin superfamily. PMID- 16263907 TI - A novel gene family in Arabidopsis encoding putative heptahelical transmembrane proteins homologous to human adiponectin receptors and progestin receptors. AB - A novel seven-transmembrane receptor family, that is comprised of human adiponectin receptors (AdipoRs) and membrane progestin receptors (mPRs) that share little sequence homology with all known G protein-coupled receptors (GPCRs), has been identified recently. Although a fish mPR has been suggested to be a GPCR, human AdipoRs seem to be structurally and functionally distinct from all known GPCRs. The identification of a novel gene family, the heptahelical protein (HHP) gene family, encoding proteins in Arabidopsis predicted to have a heptahelical transmembrane topology is reported here. There are at least five HHP genes in Arabidopsis whose encoded amino acid sequences have significant similarities to human AdipoRs and mPRs. The expression and regulation of the Arabidopsis HHP gene family has been studied here. The expression of the HHP gene family is differentially regulated by plant hormones. Steady-state levels of HHP1 mRNA are increased by treatments with abscisic acid and gibberellic acid, whereas levels of HHP2 mRNA are increased by abscisic acid and benzyladenine treatments. In addition, the expression of the HHP gene family is up-regulated by the presence of sucrose in the medium. Temperature and salt stress treatments also differentially affect the expression of the HHP genes. These novel seven transmembrane proteins previously described in yeast and animals, and now identified in plants, may represent a new class of receptors that are highly conserved across kingdoms. PMID- 16263908 TI - Physiological and molecular diversity of feather moss associative N2-fixing cyanobacteria. AB - Cyanobacteria colonizing the feather moss Pleurozium schreberi were isolated from moss samples collected in northern Sweden and subjected to physiological and molecular characterization. Morphological studies of isolated and moss-associated cyanobacteria were carried out by light microscopy. Molecular tools were used for cyanobacteria identification, and a reconstitution experiment of the association between non-associative mosses and cyanobacteria was conducted. The influence of temperature on N2 fixation in the different cyanobacterial isolates and the influence of light and temperature on N2-fixation rates in the moss were studied using the acetylene reduction assay. Two different cyanobacteria were effectively isolated from P. schreberi: Nostoc sp. and Calothrix sp. A third genus, Stigonema sp. was identified by microscopy, but could not be isolated. The Nostoc sp. was found to fix N2 at lower temperatures than Calothrix sp. Nostoc sp. and Stigonema sp. were the predominant cyanobacteria colonizing the moss. The attempt to reconstitute the association between the moss and cyanobacteria was successful. The two isolated genera of cyanobacteria in feather moss samples collected in northern Sweden differ in their temperature optima, which may have important ecological implications. PMID- 16263909 TI - KAT1 inactivates at sub-millimolar concentrations of external potassium. AB - Structural analysis of K+ channel pores suggests that the selectivity filter of the pore is an inherent sensor for extracellular K+ (Ko+); channels seem to be inactivated at low Ko+ because of a destabilization of the conducting state and a collapse of the pore. In the present study, the effect of depleting Ko+ on the activity of a plant K+ channel, KAT1, from Arabidopsis thaliana was investigated. This channel is thought to be insensitive to Ko+. The channel was therefore expressed in mammalian HEK293 cells and measured with patch clamp technology in the whole cell configuration. The effect of Ko+ depletion on channel activity was monitored from the tail currents before, during, and after washing Ko+ from the medium. The data show that a depletion of Ko+ results in a decrease in channel conductance, irrespective of whether K+ is simply removed or replaced by either Na+ or Li+. Quantitative analysis suggests that the channel has two binding sites for K+ with the dissociation constant in the order of 20 microM. This high sensitivity of the channel to Ko+ could serve as a safety mechanism, which inactivates the channel at low Ko+ and, in this way, prevents leakage of K+ from the cells via this type of channel. PMID- 16263911 TI - Oxygen isotope enrichment (delta18O) as a measure of time-averaged transpiration rate. AB - Experimental evidence is presented to show that the 18O enrichment in the leaf biomass and the mean (time-averaged) transpiration rate are positively correlated in groundnut and rice genotypes. The relationship between oxygen isotope enrichment and stomatal conductance (g(s)) was determined by altering g(s) through ABA and subsequently using contrasting genotypes of cowpea and groundnut. The Peclet model for the 18O enrichment of leaf water relative to the source water is able to predict the mean observed values well, while it cannot reproduce the full range of measured isotopic values. Further, it fails to explain the observed positive correlation between transpiration rate and 18O enrichment in leaf biomass. Transpiration rate is influenced by the prevailing environmental conditions besides the intrinsic genetic variability. As all the genotypes of both species experienced similar environmental conditions, the differences in transpiration rate could mostly be dependent on intrinsic g(s). Therefore, it appears that the delta18O of leaf biomass can be used as an effective surrogate for mean transpiration rate. Further, at a given vapour pressure difference, delta18O can serve as a measure of stomatal conductance as well. PMID- 16263912 TI - The limits of research. PMID- 16263910 TI - Increased sensitivity to salt stress in an ascorbate-deficient Arabidopsis mutant. AB - The Arabidopsis thaliana ascorbate-deficient vtc-1 mutant has only 30% ascorbate contents of the wild type (WT). This ascorbate-deficient mutant was used here to study the physiological roles of ascorbate under salt stress in vivo. Salt stress resulted in a more significant decrease in CO2 assimilatory capacity in the vtc-1 mutant than in the WT. Photosystem II function in the Arabidopsis vtc-1 mutant also showed an increased sensitivity to salt stress. Oxidative stress, indicated by the hydrogen peroxide content, increased more dramatically in the vtc-1 mutant than in the WT under salt stress. To clarify the reason for the increased oxidative stress in the vtc-1 mutant, the contents of small antioxidant compounds and the activities of several antioxidant enzymes in the ascorbate-glutathione cycle were measured. Despite an elevated glutathione pool in the vtc-1 mutant, the ascorbate contents and the reduced form of ascorbate decreased very rapidly under salt stress. These results showed that the activities of MDAR and DHAR were lower in the vtc-1 mutant than in the WT under salt stress. Thus, low intrinsic ascorbate and an impaired ascorbate-glutathione cycle in the vtc-1 mutant under salt stress probably induced a dramatic decrease in the reduced form of ascorbate, which resulted in both enhanced ROS contents and decreased NPQ in the vtc-1 mutant. PMID- 16263913 TI - Investigating the "self" in deliberate self-harm. AB - In this study, the authors explored how a group of young people aged 16 to 26 years (who identified themselves as having engaged in deliberate self-harm) made sense of the self by conducting two online focus groups and four e-mail interviews. They analyzed data using interpretive phenomenological analysis. The concept of validation was the primary means of making sense of the self and concerned the desire to be considered legitimate and of worth. This desire was clearly evident across three realms of conflict: (a) the intrinsic or extrinsic self, which marked the distinction between objective fact and subjective opinion; (b) the accepted or denied self; and (c) the notion of normality. It is possible that having one's denied self validated online might lead to an exacerbation of an individual's self-harming behavior. Further work is needed to explore the effects of online discussion forums on such taboo forms of behavior. PMID- 16263914 TI - Patient participation in health research: research with and for people with spinal cord injuries. AB - Traditionally, patients are rarely seen as partners in health research; their influence on priority setting, research design, the undertaking of research, and interpretation and dissemination of findings was marginal. Nowadays, health researchers, funding agencies, governments, and patient organizations are beginning to acknowledge that the passive role of patients in health research is no longer satisfactory. The emerging commitment and consensus concerning the aims and features of patient participation in research have created a need for an appropriate method to engage patients as partners in health research. In this article, the author argues that a responsive-constructivist approach to evaluation fits with the aims and features of patient participation. She illustrates its potential with a case example that concerns a dialogue among patients and (clinical) researchers in the field of spinal cord injuries. She reflects on learning experiences and the (un)expected difficulties and potentials in creating social conditions for patient participation in health research. PMID- 16263915 TI - Mindfulness in hospice care: practicing meditation-in-action. AB - In this interpretive study, the authors explore the experience of mindfulness among hospice caregivers who regularly practice mindfulness meditation at a Zen hospice. They explore meditative awareness constituted within themes of meditation-in-action, abiding in liminal spaces, seeing differently, and resting in groundlessness. By opening into nonconceptual, paradoxical, and uncertain dimensions of experience, hospice caregivers cultivate internal and external environments in which direct experience is increasingly held without judgment. This inquiry points to in-between spaces of human experience wherein mindfulness fosters openness and supports letting go, and creating spaces for whatever is happening in attending the living-and-dying process. PMID- 16263916 TI - Young women as smokers and nonsmokers: a qualitative social identity approach. AB - The authors used a social identity perspective to explore young women's perceptions of smoking. They carried out 13 focus groups and 6 intercept interviews with women aged 16 to 28 years in regards to the social identities that might influence young women's smoking behavior. Three identities emerged: the cool smoker applied to the initiation of smoking; considerate smokers, who were older addicted smokers; and the actual and anticipated good mother identity, which applied to young women who quit smoking during pregnancy. These identities add to our understanding of the meaning of smoking within the lives of young women and might allow more focused initiatives with this group to prevent the progression to regular addicted smoking. PMID- 16263917 TI - Maintaining support in people with paralysis: what works? AB - Social support is a protective factor for well-being in the risk-and-resilience framework, yet people with paralysis report lower levels of support compared to people without paralysis. Rather than examine deficits, in this study, the authors conducted in-depth interviews with individuals who report high levels of social support to examine what sustains this protective factor. Because relationship equity affects social support, the authors also examined this. They selected participants who reported high levels of support from a survey sample of 299 U.S. adults experiencing some form of paralysis. Seventeen participants completed the in-depth interview. The importance of reciprocity, maintaining autonomy, and a positive outlook for sustaining support were themes identified in the content analysis. In their responses, people with high support emphasized that they do all they can to affect their environment positively, so that ideally, the only assistance that they cannot provide themselves is successfully obtained from others. PMID- 16263918 TI - Information sharing in interdisciplinary team meetings: an evaluation of hospice goals. AB - The author observed interdisciplinary team (IDT) meetings of "Town Hospice" to examine how the company goal of providing for the "psychosocial needs of the patient and their loved ones" is addressed. More specifically, she used an ethnographic approach to explore how case managers negotiate the addition of psychosocial information about patients during IDT meetings. She found that psychosocial information on patients was primarily limited to three types of information sharing: (a) information related to care goals, (b) family issues related to bereavement and caretaking, and (c) the request of additional help from team members. Furthermore, she understood that the addition of psychosocial information creates a dialectical tension for the team. PMID- 16263919 TI - An introduction to concept mapping as a participatory public health research method. AB - In this article, the authors introduce concept mapping as a useful participatory research method for public health researchers interested in generating hypotheses and developing theory. The authors first provide an overview of concept mapping, which combines qualitative approaches with quantitative analytical tools to produce visual displays of the relationship between ideas. Then, they present an illustrative research application of the method to the exploration of women's perceptions of the relationship between residential neighborhood factors and intimate partner violence experiences. They give attention to the data collection and analysis procedures and to demonstrating the intricacies of using concept mapping for public health research purposes. Finally, the article concludes with a discussion of the unique contributions and challenges associated with concept mapping. PMID- 16263921 TI - Infection patterns in barley and wheat spikes inoculated with wild-type and trichodiene synthase gene disrupted Fusarium graminearum. AB - Fusarium head blight epidemics of wheat and barley cause heavy economic losses to farmers due to yield decreases and production of mycotoxin that renders the grain useless for flour and malt products. No highly resistant cultivars are available at present. Hyphae of germinating fungal spores use different paths of infection: After germination at the extruded tip of an ovary, the hyphae travel along the epicarp in the space between the lemma and palea. Infection of the developing kernel proceeds through the epicarp, successively destroying the layers of the fruit coat and finally the starch and protein accumulating endosperm. Hyphae reaching the rachis proceed to apically located developing kernels. Using a constitutively green fluorescence protein-expressing Fusarium wild-type strain, and its knockout mutant, preventing trichothecene synthesis, we demonstrate that trichothecenes are not a virulence factor during infection through the fruit coat. In the absence of trichothecenes, the fungus is blocked by the development of heavy cell wall thickenings in the rachis node of Nandu wheat, a defense inhibited by the mycotoxin. In barley hyphae of both wild-type and the trichothecene knockout mutant, are inhibited at the rachis node and rachilla, limiting infection of adjacent florets through the phloem and along the surface of the rachis. Effective resistance to Fusarium head blight requires expression of genes that combat these different pathways of infection. PMID- 16263922 TI - Expanding the repertoire of the eukaryotic selenoproteome. PMID- 16263923 TI - Chemical imaging of tissue in vivo with video-rate coherent anti-Stokes Raman scattering microscopy. AB - Imaging living organisms with molecular selectivity typically requires the introduction of specific labels. Many applications in biology and medicine, however, would significantly benefit from a noninvasive imaging technique that circumvents such exogenous probes. In vivo microscopy based on vibrational spectroscopic contrast offers a unique approach for visualizing tissue architecture with molecular specificity. We have developed a sensitive technique for vibrational imaging of tissues by combining coherent anti-Stokes Raman scattering (CARS) with video-rate microscopy. Backscattering of the intense forward-propagating CARS radiation in tissue gives rise to a strong epi-CARS signal that makes in vivo imaging possible. This substantially large signal allows for real-time monitoring of dynamic processes, such as the diffusion of chemical compounds, in tissues. By tuning into the CH(2) stretching vibrational band, we demonstrate CARS imaging and spectroscopy of lipid-rich tissue structures in the skin of a live mouse, including sebaceous glands, corneocytes, and adipocytes, with unprecedented contrast at subcellular resolution. PMID- 16263924 TI - Development of spectral and temporal response selectivity in the auditory cortex. AB - The mechanisms by which hearing selectivity is elaborated and refined in early development are very incompletely determined. In this study, we documented contributions of progressively maturing inhibitory influences on the refinement of spectral and temporal response properties in the primary auditory cortex. Inhibitory receptive fields (IRFs) of infant rat auditory cortical neurons were spectrally far broader and had extended over far longer duration than did those of adults. The selective refinement of IRFs was delayed relative to that of excitatory receptive fields by an approximately 2-week period that corresponded to the critical period for plasticity. Local application of a GABA(A) receptor antagonist revealed that intracortical inhibition contributes to this progressive receptive field maturation for response selectivity in frequency. Conversely, it had no effect on the duration of IRFs or successive-signal cortical response recovery times. The importance of exposure to patterned acoustic inputs was suggested when both spectral and temporal IRF maturation were disrupted in rat pups reared in continuous, moderate-intensity noise. They were subsequently renormalized when animals were returned to standard housing conditions as adults. PMID- 16263925 TI - Modeling lateral geniculate nucleus cell response spectra and Munsell reflectance spectra with cone sensitivity curves. AB - We find that the cell response spectra of lateral geniculate nucleus cells, as well as the reflectance spectra of Munsell color chips, may be modeled by using the cone sensitivity functions of the long and medium cones. We propose a simple model for how the neural signals from the photoreceptors might be combined in the retina to closely approximate the reflectance spectra of Munsell color chips without input from the short cone. PMID- 16263927 TI - Profile of Aziz Sancar. PMID- 16263929 TI - Quantitative NMR spectroscopy of supramolecular complexes: dynamic side pores in ClpP are important for product release. AB - The highly conserved, 300-kDa cylindrical protease ClpP is an important component of the cellular protein quality machinery. It consists of 14 subunits arranged into two heptameric rings that enclose a large chamber containing the protease active sites. ClpP associates with ClpX and ClpA ATPases that unfold and translocate substrates into the protease catalytic chamber through axial pores located at both ends of the ClpP cylinder. Although the pathway of substrate delivery is well established, the pathway of product release is unknown. Here, we use recently developed transverse relaxation optimized spectroscopy (TROSY) of methyl groups to show that the interface between the heptameric rings exchanges between two structurally distinct conformations. The conformational exchange process has been quantified by magnetization exchange and methyl TROSY relaxation dispersion experiments recorded between 0.5 degrees C and 40 degrees C, so that the thermodynamic properties for the transition could be obtained. Restriction of the observed motional freedom in ClpP through the introduction of a cysteine linkage results in a protease where substrate release becomes significantly slowed relative to the rate observed in the reduced enzyme, suggesting that the observed motions lead to the formation of transient side pores that may play an important role in product release. PMID- 16263928 TI - Glutathionylation of chloroplast thioredoxin f is a redox signaling mechanism in plants. AB - Thioredoxin f (TRXf) is a key factor in the redox regulation of chloroplastic carbon fixation enzymes, whereas glutathione is an important thiol buffer whose status is modulated by stress conditions. Here, we report specific glutathionylation of TRXf. A conserved cysteine is present in the TRXf primary sequence, in addition to its two active-site cysteines. The additional cysteine becomes glutathionylated when TRXf is exposed to oxidized glutathione or to reduced glutathione plus oxidants. No other chloroplastic TRX, from either Arabidopsis or Chlamydomonas, is glutathionylated under these conditions. Glutathionylation decreases the ability of TRXf to be reduced by ferredoxin thioredoxin reductase and results in impaired light activation of target enzymes in a reconstituted thylakoid system. Although several mammalian proteins undergoing glutathionylation have already been identified, TRXf is among the first plant proteins found to undergo this posttranslational modification. This report suggests that a crosstalk between the TRX and glutathione systems mediates a previously uncharacterized form of redox signaling in plants in stress conditions. PMID- 16263930 TI - Phosphoinositol lipids bind to phosphatidylinositol 3 (PI3)-kinase enhancer GTPase and mediate its stimulatory effect on PI3-kinase and Akt signalings. AB - Phosphatidylinositol 3 (PI3)-kinase enhancer (PIKE) is a nuclear GTPase that enhances PI3-kinase activity in a GTP-dependent manner. Both PIKE-L and -A isoforms contain GTPase, pleckstrin homology (PH), ADP ribosylation factor-GTPase activating protein, and two ankyrin repeats domains, and C-terminal ADP ribosylation factor-GTPase-activating protein activates its internal GTPase activity. However, whether PH domain modulates the intramolecular action and subsequently influences its downstream signalings remains elusive. Here we show that PH domain from PIKE-L robustly binds PI(3,4,5)P(3) and exclusively resides in the nucleus. By contrast, the mutant (K679,687N), unable to bind phosphoinositol lipids, translocates to the cytoplasm. This mutation substantially compromises the stimulatory effects on PI3-kinase by PIKE-L. Surprisingly, PH domain from PIKE-A distributes in the cytoplasm. Similar mutation in PH domain of PIKE-A abolishes its binding to PI(3,4,5)P(3) and significantly decreases its activation of Akt. Moreover, amplified PIKE-A from human cancers contains mutations and highly stimulates Akt kinase activity, correlating with its GTPase activity. Thus, phosphatidylinositols regulate PIKE GTPase activity, mediating its downstream PI3-kinase/Akt signaling through a feedback mechanism by binding to its PH domain. PMID- 16263931 TI - Ultra-high-yield growth of vertical single-walled carbon nanotubes: Hidden roles of hydrogen and oxygen. AB - An oxygen-assisted hydrocarbon chemical vapor deposition method is developed to afford large-scale, highly reproducible, ultra-high-yield growth of vertical single-walled carbon nanotubes (V-SWNTs). It is revealed that reactive hydrogen species, inevitable in hydrocarbon-based growth, are damaging to the formation of sp(2)-like SWNTs in a diameter-dependent manner. The addition of oxygen scavenges H species and provides a powerful control over the C/H ratio to favor SWNT growth. The revelation of the roles played by hydrogen and oxygen leads to a unified and universal optimum-growth condition for SWNTs. Further, a versatile method is developed to form V-SWNT films on any substrate, lifting a major substrate-type limitation for aligned SWNTs. PMID- 16263932 TI - The amyloid stretch hypothesis: recruiting proteins toward the dark side. AB - A detailed understanding of the molecular events underlying the conversion and self-association of normally soluble proteins into amyloid fibrils is fundamental to the identification of therapeutic strategies to prevent or cure amyloid related disorders. Recent investigations indicate that amyloid fibril formation is not just a general property of the polypeptide backbone depending on external factors, but that it is strongly modulated by amino acid side chains. Here, we propose and address the validation of the premise that the amyloidogenicity of a protein is indeed localized in short protein stretches (amyloid stretch hypothesis). We demonstrate that the conversion of a soluble nonamyloidogenic protein into an amyloidogenic prone molecule can be triggered by a nondestabilizing six-residue amyloidogenic insertion in a particular structural environment. Interestingly enough, although the inserted amyloid sequences clearly cause the process, the protease-resistant core of the fiber also includes short adjacent sequences from the otherwise soluble globular domain. Thus, short amyloid stretches accessible for intermolecular interactions trigger the self assembly reaction and pull the rest of the protein into the fibrillar aggregate. The reliable identification of such amyloidogenic stretches in proteins opens the possibility of using them as targets for the inhibition of the amyloid fibril formation process. PMID- 16263933 TI - Targeted down-regulation of cytochrome P450 enzymes for forage quality improvement in alfalfa (Medicago sativa L.). AB - Improving the digestibility of forages provides a means to enhance animal performance and protect the environment against excessive animal waste. Increased lignin content during maturity, and corresponding changes in lignin composition, correlate with decreased digestibility of forages. These relationships have yet to be investigated in isogenic systems. By targeting three specific cytochrome P450 enzymes of the lignin pathway for antisense down-regulation, we generated transgenic alfalfa lines with a range of differences in lignin content and composition. There was a strong negative relationship between lignin content and rumen digestibility, but no relationship between lignin composition and digestibility, in these transgenic lines. Models for genetic manipulation of forage digestibility based on the changes in lignin composition that increase paper-pulping efficiency in trees are therefore invalid. Down-regulation of 4 coumarate 3-hydroxylase provided the largest improvements in digestibility yet seen in a forage crop. PMID- 16263934 TI - Unconscious processing of orientation and color without primary visual cortex. AB - In humans, the primary visual cortex (V1) is essential for conscious vision. However, even without V1 and in the absence of awareness, some preserved ability to accurately respond to visual inputs has been demonstrated, a phenomenon referred to as blindsight. We used transcranial magnetic stimulation (TMS) to deactivate V1, producing transient blindness for visual targets presented in a foveal, TMS-induced scotoma. Despite unawareness of these targets, performance on forced choice discrimination tasks for orientation (experiment 1) and color (experiment 2) were both significantly above chance. In addition to demonstrating that TMS can be successfully used to induce blindsight within a normal population, these results suggest a functioning geniculoextrastriate visual pathway that bypasses V1 and can process orientation and color in the absence of conscious awareness. PMID- 16263926 TI - Simple sequence repeat-based consensus linkage map of Bombyx mori. AB - We established a genetic linkage map employing 518 simple sequence repeat (SSR, or microsatellite) markers for Bombyx mori (silkworm), the economically and culturally important lepidopteran insect, as part of an international genomics program. A survey of six representative silkworm strains using 2,500 (CA)n- and (CT)n-based SSR markers revealed 17-24% polymorphism, indicating a high degree of homozygosity resulting from a long history of inbreeding. Twenty-nine SSR linkage groups were established in well characterized Dazao and C108 strains based on genotyping of 189 backcross progeny derived from an F(1) male mated with a C108 female. The clustering was further focused to 28 groups by genotyping 22 backcross progeny derived from an F(1) female mated with a C108 male. This set of SSR linkage groups was further assigned to the 28 chromosomes (established linkage groups) of silkworm aided by visible mutations and cleaved amplified polymorphic sequence markers developed from previously mapped genes, cDNA sequences, and cloned random amplified polymorphic DNAs. By integrating a visible mutation p (plain, larval marking) and 29 well conserved genes of insects onto this SSR-based linkage map, a second generation consensus silkworm genetic map with a range of 7-40 markers per linkage group and a total map length of approximately 3431.9 cM was constructed and its high efficiency for genotyping and potential application for synteny studies of Lepidoptera and other insects was demonstrated. PMID- 16263935 TI - Expression of a calmodulin-binding KCNQ2 potassium channel fragment modulates neuronal M-current and membrane excitability. AB - KCNQ2 and KCNQ3 ion channel pore-forming subunits coassemble to form a heteromeric voltage-gated potassium channel that underlies the neuronal M current. We and others showed that calmodulin (CaM) binds to specific sequence motifs in the C-terminal domain of KCNQ2 and KCNQ3. We also found that a fusion protein containing a KCNQ2 CaM-binding motif, coexpressed with KCNQ2 and KCNQ3, competes with the full-length KCNQ2 channel for CaM binding and thereby decreases KCNQ2/3 current density in heterologous cells. We have explored the importance of CaM binding for the generation of the native M-current and regulation of membrane excitability in rat hippocampal neurons in primary cell culture. M-current properties were studied in cultured neurons by using whole-cell patch clamp recording. The M-current density is lower in neurons expressing the CaM-binding motif fusion protein, as compared to control neurons transfected with vector alone. In contrast, no change in M-current density is observed in cells transfected with a mutant fusion protein that is unable to bind CaM. The CaM binding fusion protein does not influence the rapidly inactivating A-current or the large conductance calcium-activated potassium channel-mediated fast spike afterhyperpolarization in neurons in which the M-current is suppressed. Furthermore, the CaM-binding fusion protein, but not the nonbinding mutant, increases both the number of action potentials evoked by membrane depolarization and the size of the spike afterdepolarization. These results suggest that CaM binding regulates M-channel function and membrane excitability in the native neuronal environment. PMID- 16263936 TI - Determination of cell survival by RING-mediated regulation of inhibitor of apoptosis (IAP) protein abundance. AB - Inhibitor of apoptosis (IAP) proteins, which bind to caspases via their baculoviral IAP repeat domains, also bear RING domains that enable them to promote ubiquitylation of themselves and other interacting proteins. Here we show that the RING domain of cIAP1 allows it to bind directly to the RING of X-linked IAP, causing its ubiquitylation and degradation by the proteasome, thus revealing a mechanism by which IAPs can regulate their abundance. Expression of a construct containing the RING of cellular IAP1 was able to deplete melanoma cells of endogenous X-linked IAP, promoted apoptosis, and also markedly reduced their clonogenicity when treated with cisplatin. Cross control of protein levels by RING domains may therefore enable their levels to be manipulated therapeutically. PMID- 16263937 TI - Agriculture in the developing world: Connecting innovations in plant research to downstream applications. AB - Enhancing agricultural productivity in those areas of the world bypassed by the Green Revolution will require new approaches that provide incentives and funding mechanisms that promote the translation of new innovations in plant science into concrete benefits for poor farmers. Through better dialogue, plant breeders and laboratory scientists from both the public and private-sectors need to find solutions for the key constraints to crop production, many of which center around abiotic and biotic stresses. The revolution in plant genomics has opened up new perspectives and opportunities for plant breeders who can now apply molecular markers to assess and enhance diversity in their germplasm collections, to introgress valuable traits from new sources, and to identify genes that control key traits. Functional genomics is also providing another powerful route to the identification of such genes. The ability to introgress beneficial genes under the control of specific promoters through transgenic approaches is yet one more stepping stone in the path to targeted approaches to crop improvement, and the new sciences have identified a vast array of genes that have exciting potential for crop improvement. For a few crops with viable markets, such as maize and cotton, some of the traits developed by the private sector are already showing benefits for farmers of the developing world, but the public sector will need to develop new skills and overcome a number of hurdles to carry out similar efforts for other crops and traits useful to very poor farmers. PMID- 16263938 TI - Impact of HIF-1alpha and HIF-2alpha on proliferation and migration of human pulmonary artery fibroblasts in hypoxia. AB - Proliferation of adventitial fibroblasts of small intrapulmonary arteries (FBPA) has been disclosed as an early event in the development of pulmonary hypertension and cor pulmonale in response to hypoxia. We investigated the role of hypoxia inducible transcription factors (HIF) in human FBPA exposed to hypoxia. Primary cultures of FBPA displayed a strong mitogenic response to 24 h hypoxia, whereas the rate of apoptosis was significantly suppressed. In addition, the migration of FBPA was strongly increased under hypoxic conditions but not the expression of alpha-smooth muscle actin. Hypoxia induced a marked up-regulation (protein level) of both HIF-1alpha and HIF-2alpha, alongside with nuclear translocation of these transcription factors. Specific inhibition of either HIF-1alpha or HIF-2alpha was achieved by RNA interference technology, as proven by HIF-1alpha and HIF-2alpha mRNA and protein analysis and expression analysis of HIF downstream target genes. With the use of this approach, the hypoxia-induced proliferative response of the FBPA was found to be solely HIF-2alpha dependent, whereas the migratory response was significantly reduced by both HIF-1alpha and HIF-2alpha interference. In conclusion, HIF up-regulation is essential for hypoxic cellular responses in human pulmonary artery adventitial fibroblasts such as proliferation and migration, mimicking the pulmonary hypertensive phenotype in vivo. Differential HIF subtype dependency was noted, with HIF-2alpha playing a predominant role, which may offer future intervention strategies. PMID- 16263939 TI - Transthyretin knockouts are a new mouse model for increased neuropeptide Y. AB - Transthyretin (TTR) has access to the brain and nerve through the blood and cerebrospinal fluid. To investigate TTR function in nervous system homeostasis, differential gene expression in wild-type (WT) and TTR knockout (KO) mice was assessed. Peptidylglycine alpha-amidating monooxygenase (PAM), the rate-limiting enzyme in neuropeptide maturation, is overexpressed in the peripheral (PNS) and central nervous system (CNS) of TTR KOs that, consequently, display increased neuropeptide Y (NPY) levels. NPY acts on energy homeostasis by increasing white adipose tissue lipoprotein lipase (LPL) and decreasing thermogenesis; accordingly, we show increased LPL expression and activity in white adipose tissue, PNS, and CNS as well as decreased body temperature in TTR KOs. Associated to increased NPY levels, TTR KOs display increased carbohydrate consumption and preference. In neuronal cells, absence of TTR is related to increased PAM activity, NPY levels and LPL expression, reinforcing that TTR is involved in neuropeptide maturation and that increased NPY correlates with LPL overexpression in the nervous system. Furthermore, we provide molecular insights to the reduced depressive behavior of TTR KOs, as NPY is anti-depressant. Our findings demonstrate that TTR KOs are a model for increased NPY and that TTR plays a role in nervous system physiology. PMID- 16263941 TI - Wheeze, asthma diagnosis and medication use in developing countries. PMID- 16263940 TI - c-Flip expression and function in fetal mouse gonocytes. AB - Apoptosis is a key mechanism in spermatogenesis, and in testis, most gonocytes degenerate at fetal and postnatal ages to select a cell subset committed to become germ stem cells. The aim of the present study is to investigate mechanisms controlling the massive apoptosis of fetal gonocytes. We evaluated the expression and function of c-Flip, an apoptosis inhibitor known to interfere with the proapoptotic Fas-signaling pathway in a variety of cell types, but never investigated in fetal testis. Expression of c-Flip long isoform (c-FlipL) within fetal testis was localized in gonocytes at 16.5 and 18.5 days post coitum (dpc), both at the mRNA and protein level, while it was weakly expressed or undetectable at earlier stages. Moreover, Fas protein was found in fetal testes at 13.5, 16.5, and 18.5 dpc. Testes at 18.5 dpc, expressing high levels of c-FlipL, were resistant to Fas-induced apoptosis while they became highly sensitive when c FlipL was inhibited by antisense c-Flip oligos. In addition, there was an inverse relation between gonocyte spontaneous apoptosis sensitivity and c-FlipL levels. Furthermore, caspase-10 activity was inversely related with c-FlipL expression, suggesting that caspase-10 might be a target of c-FlipL. These data represent the first evidence demonstrating c-Flip expression in fetal testes and its role in protecting gonocytes from Fas-dependent apoptosis. PMID- 16263942 TI - Exercise in COPD: damned if you do and damned if you don't. PMID- 16263943 TI - Exercise training and inspiratory muscle training in patients with bronchiectasis. PMID- 16263944 TI - Usefulness of transbronchial needle aspiration in evaluating patients with lung cancer. PMID- 16263945 TI - Orphan lung diseases in childhood: still unadopted? PMID- 16263947 TI - Wheeze, asthma diagnosis and medication use: a national adult survey in a developing country. AB - BACKGROUND: As relatively little is known about adult wheeze and asthma in developing countries, this study aimed to determine the predictors of wheeze, asthma diagnosis, and current treatment in a national survey of South African adults. METHODS: A stratified national probability sample of households was drawn and all adults (>14 years) in the selected households were interviewed. Outcomes of interest were recent wheeze, asthma diagnosis, and current use of asthma medication. Predictors of interest were sex, age, household asset index, education, racial group, urban residence, medical insurance, domestic exposure to smoky fuels, occupational exposure, smoking, body mass index, and past tuberculosis. RESULTS: A total of 5671 men and 8155 women were studied. Although recent wheeze was reported by 14.4% of men and 17.6% of women and asthma diagnosis by 3.7% of men and 3.8% of women, women were less likely than men to be on current treatment (OR 0.6; 95% confidence interval (CI) 0.5 to 0.8). A history of tuberculosis was an independent predictor of both recent wheeze (OR 3.4; 95% CI 2.5 to 4.7) and asthma diagnosis (OR 2.2; 95% CI 1.5 to 3.2), as was occupational exposure (wheeze: OR 1.8; 95% CI 1.5 to 2.0; asthma diagnosis: OR 1.9; 95% CI 1.4 to 2.4). Smoking was associated with wheeze but not asthma diagnosis. Obesity showed an association with wheeze only in younger women. Both wheeze and asthma diagnosis were more prevalent in those with less education but had no association with the asset index. Independently, having medical insurance was associated with a higher prevalence of diagnosis. CONCLUSIONS: Some of the findings may be to due to reporting bias and heterogeneity of the categories wheeze and asthma diagnosis, which may overlap with post tuberculous airways obstruction and chronic obstructive pulmonary disease due to smoking and occupational exposures. The results underline the importance of controlling tuberculosis and occupational exposures as well as smoking in reducing chronic respiratory morbidity. Validation of the asthma questionnaire in this setting and research into the pathophysiology of post tuberculous airways obstruction are also needed. PMID- 16263952 TI - Klippel-Trenaunay syndrome with small vessel pulmonary arterial hypertension. AB - A patient with Klippel-Trenaunay syndrome and pulmonary arterial hypertension not associated with chronic thrombo-embolic pulmonary hypertension is described. It is hypothesised that pulmonary arterial hypertension is another complication associated with the Klippel-Trenaunay syndrome, possibly due to haemodynamic changes of small vessel abnormalities. PMID- 16263954 TI - Late CF caused by homozygous IVS8-5T CFTR polymorphism. PMID- 16263955 TI - TIMP-3 promoter gene polymorphisms in BFL. PMID- 16263956 TI - Palmoplantar keratoderma ("tripe palms") associated with primary pulmonary adenocarcinoma. PMID- 16263957 TI - A farewell message from the editor. PMID- 16263959 TI - Professors Toshihide Yamashita and Zhigang He to receive 2005 Ameritec Prize for paralysis research. PMID- 16263960 TI - Rehabilitation and functional neuroimaging dose-response trajectories for clinical trials. AB - BACKGROUND: In clinical trials, behavioral outcomes and physiological measures of activity-dependent plasticity that evolve with task-oriented therapies may fail to reach statistical significance. When significant, clinical effectiveness may not be robust enough to alter professional practices. OBJECTIVE: Provide the conceptual basis for a research design to optimize the effect of an experimental treatment. METHODS: Literature review. RESULTS: Research designs usually do not take into consideration the dynamic state of each subject's potential responsiveness to an intervention. Providing a rational, rather than convenient, intensity and duration of therapy may remedy this potential confounder for clinical trials. To determine whether a most effective dose of a therapy exists, investigators could assess subjects before the intervention, administer interim measures at planned intervals, and continue the intervention until the primary behavioral outcomes or functional imaging parameters or both reach a plateau for at least 15 h of additional treatment. CONCLUSION: Promising interventions ought to be continued in phase II/III trials until subjects reach an asymptote in the primary outcome for behavioral gains. For neuroimaging studies that aim to correlate brain-behavior measures during rehabilitation, the specific intervention should also continue until behavioral gains and cerebral adaptations have attained a persistent plateau. Future trials can investigate whether functional neuroimaging performed in parallel with repeated behavioral assessments can better inform researchers about the optimal duration of an experimental therapy and a subject's maximal capacity for intervention-induced cerebral reorganization. PMID- 16263961 TI - License to run: exercise impacts functional plasticity in the intact and injured central nervous system by using neurotrophins. AB - Exercise has been found to impact molecular systems important for maintaining neural function and plasticity. A characteristic finding for the effects of exercise in the brain and spinal cord has been the up-regulation of brain-derived neurotrophic factor (BDNF). This review focuses on the ability of exercise to impact brain circuitry by promoting neuronal repair and enhance learning and memory by increasing neurotrophic support. A paragon for the role of activity dependent neurotrophins in the CNS is the capacity of BDNF to facilitate synaptic function and neuronal excitability. The authors discuss the effects of exercise in the intact and injured brain and spinal cord injury and the implementation of exercise preinjury and postinjury. As the CNS displays a capacity for plasticity throughout one's lifespan, exercise may be a powerful lifestyle implementation that could be used to augment synaptic plasticity, promote behavioral rehabilitation, and counteract the deleterious effects of aging. PMID- 16263962 TI - Combined effects of neurotrophin secreting transplants, exercise, and serotonergic drug challenge improve function in spinal rats. AB - OBJECTIVES: To determine the effects of neurotrophin-secreting transplants combined with exercise and serotonergic drug challenges on recovery of hindlimb function in rats with midthoracic spinal cord transection injuries. METHODS: Spinalized animals received transplants of fibroblasts genetically modified to express brain-derived neurotrophic factor and neurotrophin-3 and daily cycling exercise. Hindlimb movement in an open-field test (BBB) was scored weekly. Serotonin agonists were used monthly to further stimulate motor function. Axonal growth was quantified in the transplant and at L5 using immunocytochemical markers. Weights of hindlimb muscles were used to assess muscle atrophy. RESULTS: Neurotrophin-secreting transplants stimulated axonal growth, and cycling prevented muscle atrophy, but individual treatments did not improve motor scores. Combined treatments resulted in improvements in motor function. Serotonergic agonists further improved function in all groups, and transplant groups with exercise achieved weight-supporting levels following drug treatment. CONCLUSION: Combined treatments, but not individual treatments, improved hindlimb function. PMID- 16263963 TI - Changes in supraspinal activation patterns following robotic locomotor therapy in motor-incomplete spinal cord injury. AB - OBJECTIVES: Body weight-supported treadmill training (BWSTT) is a task-specific rehabilitation strategy that enhances functional locomotion in patients following spinal cord injury (SCI). Supraspinal centers may play an important role in the recovery of over-ground locomotor function in patients with motor-incomplete SCI. The purpose of this study was to evaluate the potential for supraspinal reorganization associated with 12 weeks of robotic BWSTT using functional magnetic resonance imaging (fMRI). METHODS: Four men with motor-incomplete SCI participated in this study. Time since onset ranged from 14 weeks to 48 months post-SCI injury. All subjects were trained with BWSTT 3 times weekly for 12 weeks. This training was preceded and followed by fMRI study of supraspinal activity during a movement task. Testing of locomotor disability included the Walking Index for Spinal Cord Injury (WISCI II) and over-ground gait speed. RESULTS: All subjects demonstrated some degree of change in the blood-oxygen level-dependent (BOLD) signal following BWSTT. fMRI results demonstrated greater activation in sensorimotor cortical regions (S1, S2) and cerebellar regions following BWSTT. CONCLUSIONS: Intensive task-specific rehabilitative training, such as robotic BWSTT, can promote supraspinal plasticity in the motor centers known to be involved in locomotion. Furthermore, improvement in over-ground locomotion is accompanied by an increased activation of the cerebellum. PMID- 16263964 TI - The effect of different physiological stimuli on skin vasomotor reflexes above and below the lesion in human chronic spinal cord injury. AB - OBJECTIVE: Spinal cord injury (SCI) results in disruption of descending tonic activation of sympathetic circuits in the spinal cord. The authors determined whether different stimuli that increase sympathetic activity induced cutaneous vasoconstriction (skin vasomotor reflex, SkVR) above and below the level of lesion in subjects with clinically complete SCI. METHODS: Baseline skin blood flow (SkBF) and SkVR reduction rate in the pulp of the finger and great toe was measured by laser Doppler probes in chronic complete SCI and in controls. RESULTS: In the finger, baseline SkBF was similar in SCI and controls. The SkVR was elicited by all stimuli in controls but was significantly diminished to gasp, mental arithmetic, tactile stimulation, cutaneous cold, and deep breathing in high SCI compared to controls. In the toe, baseline SkBF was less stable in both controls and SCI. SkVR trends were identified in controls, and responses were not present or greatly reduced in high and low SCI. CONCLUSIONS: Measurements of skin vasomotor reflexes to physiological stimuli may be a noninvasive method to evaluate the extent of sympathetic adrenergic pathways in chronic SCI. This is of clinical relevance in monitoring recovery of sympathetic adrenergic function either spontaneously or following repair interventions. PMID- 16263965 TI - Long-term follow-up of patients with spinal cord injury. AB - OBJECTIVE: The aim of this study was to evaluate the motor, sensory, and functional recovery in patients with spinal cord injury (SCI). METHODS: Forty-one patients with SCI participated in this study. Twenty patients were evaluated after discharge. Each patient was evaluated by the American Spinal Injury Association (ASIA) impairment scale and the Functional Independence Measure (FIM) at admission, before discharge, and at least at 6 months after discharge. Friedman, Dunn, and Mann-Whitney U tests were used for statistical analysis. RESULTS: There were 17 male and 3 female patients. Seven patients had complete SCI, and 13 patients had incomplete SCI. The evaluation of motor, sensory, and FIM scores at admission showed significant improvement in all of the patients during the follow-up period (P < 0.0001). Five incompletely injured cases improved with regard to ASIA staging. Motor and FIM scores significantly increased at follow-up for converted and unconverted patients. All parameters increased at follow-up in patients who were complete and incomplete. Motor scores significantly increased at discharge and at follow-up. FIM scores also increased significantly at follow-up in incomplete patients. CONCLUSION: Motor, sensory, and FIM scores increased in patients with SCI after a follow-up period of 18 months. Improvement to a higher ASIA stage could be accomplished by 25% of the patients. Although both complete and incomplete patients recovered significantly at the follow-up period, only incompletely injured cases could convert to a higher ASIA stage. PMID- 16263966 TI - Impaired grip force modulation in the ipsilesional hand after unilateral middle cerebral artery stroke. AB - Understanding grasping control after stroke is important for relearning motor skills. The authors examined 10 individuals (5 males; 5 females; ages 32-86) with chronic unilateral middle cerebral artery (MCA) stroke (4 right lesions; 6 left lesions) when lifting a novel test object using skilled precision grip with their ipsilesional ("unaffected") hand compared to healthy controls (n = 14; 6 males; 8 females; ages 19-86). All subjects possessed normal range of motion, cutaneous sensation, and proprioception in the hand tested and had no apraxia or cognitive deficits. Subjects lifted the object 10 times at each object weight (260 g, 500 g, 780 g) using a moderately paced self-selected lifting speed. The normal horizontal ("grip") force and vertical tangential ("lift") force were separately measured at the thumb and index finger. Regardless of the object weight or stroke location, the stroke group generated greater grip forces at liftoff of the object (> or =39%; P < or = 0.05) and across the dynamic (P < or = 0.05) and static portions (P < or = 0.05) of the lifts compared to the healthy group. Peak lift forces were equivalent between groups, suggesting accurate load force information processing occurred. These results warrant further investigation of altered sensorimotor processing or compensatory biomechanical strategies that may lead to inaccurate grip force execution after strokes. PMID- 16263967 TI - Development of a schedule of current physiotherapy treatment used to improve movement control and functional use of the lower limb after stroke: a precursor to a clinical trial. AB - OBJECTIVE: To develop a treatment schedule of physical therapy techniques used to improve movement control and functional use of the paretic lower limb after stroke in U.K. clinical centers to be involved in a subsequent clinical trial of experimental interventions given in addition to routine clinical practice. METHODS: Ten physiotherapists experienced in stroke rehabilitation who worked in or near the clinical centers to be involved in a subsequent clinical trial completed an individual semi-structured interview. The verbatim transcripts were condensed independently by 2 researchers into a draft list of interventions. The researchers then resolved disagreement through discussion and produced a preliminary list of interventions. At a focus group meeting, the participating physiotherapists discussed the preliminary list, refined it to produce a final list, and then transformed it into a draft treatment schedule. The draft treatment schedule was piloted in clinical practice. Refinements were made, and the final treatment schedule was produced. RESULTS: The treatment schedule consists of an A4 recording form with instructions and glossary of terms printed on the back. Each treatment record provides information including duration of treatment, treatment aims, and the 45 specific physical therapy interventions provided in the 11 sections (e.g., "splinting techniques" and "function - in sitting towards standing"). CONCLUSION: A treatment schedule was produced, which can now be used in a subsequent phase II evaluative trial. PMID- 16263973 TI - A population-based study to determine the performance of the Cognitive Adaptive Test/Clinical Linguistic and Auditory Milestone Scale to Predict the Mental Developmental Index at 18 Months on the Bayley Scales of Infant Development-II in very preterm infants. AB - OBJECTIVES: To determine optimal ages to perform the Cognitive Adaptive Test/Clinical Linguistic and Auditory Milestone Scale (CAT/CLAMS) and optimal "cutoff" score of the CAT/CLAMS to screen very preterm infants (<31 weeks) for severe cognitive-adaptive delay and to ascertain the sensitivity, specificity and likelihood ratios using optimal cutoff scores compared with the Mental Developmental Index (MDI) of the Bayley Scales of Infant Development II. METHODS: A population-based cohort of very preterm infants who were born to mothers who resided in Nova Scotia or Prince Edward Island were evaluated at 4, 8, 12, and 18 months' corrected gestational age, which included a CAT/CLAMS by a physician. At 18 months' corrected gestational age, each child was assessed using the Bayley Scales of Infant Development II, the "gold standard" for developmental delay in young infants. The results of each CAT/CLAMS was compared with the 18-month MDI to identify significant developmental delay (MDI <70). RESULTS: Optimal scores on the CAT/CLAMS to identify correctly MDI <70 were determined by using the kappa statistic for chance independent agreement. Sensitivities and specificities for optimal cutoff scores were as follows: 4-month score <109 (88% and 37%), 8-month score <98 (75% and 82%), 12-month score <81 (63% and 99%), and 18-month score <83 (88% and 98%). CONCLUSION: Sensitivity and specificity of the CAT/CLAMS are high in very preterm infants at identifying major developmental delay at 12 and 18 months. For follow-up programs without psychology services, the CAT/CLAMS at 12 and 18 months is a reasonable screening tool to determine which children need expedited psychology referral for cognitive delay. PMID- 16263972 TI - The health status of southern children: a neglected regional disparity. AB - PURPOSE: Great variations exist in child health outcomes among states in the United States, with southern states consistently ranked among the lowest in the country. Investigation of the geographical distribution of children's health status and the regional factors contributing to these outcomes has been neglected. We attempted to identify the degree to which region of residence may be linked to health outcomes for children with the specific aim of determining whether living in the southern region of the United States is adversely associated with children's health status. METHODS: A child health index (CHI) that ranked each state in the United States was computed by using state-specific composite scores generated from outcome measures for a number of indicators of child health. Five indicators for physical health were chosen (percent low birth weight infants, infant mortality rate, child death rate, teen death rate, and teen birth rates) based on their historic and routine use to define health outcomes in children. Indicators were calculated as rates or percentages. Standard scores were calculated for each state for each health indicator by subtracting the mean of the measures for all states from the observed measure for each state. Indicators related to social and economic status were considered to be variables that impact physical health, as opposed to indicators of physical health, and therefore were not used to generate the composite child health score. These variables were subsequently examined in this study as potential confounding variables. Mapping was used to redefine regional groupings of states, and parametric tests (2-sample t test, analysis of means, and analysis-of-variance F tests) were used to compare the means of the CHI scores for the regional groupings and test for statistical significance. Multiple-regression analysis computed the relationship of region, social and economic indicators, and race to the CHI. Simple linear-regression analyses were used to assess the individual effect of each indicator. RESULTS: A geographic region of contiguous states, characterized by their poor child health outcomes relative to other states and regions of the United States, exists within the "Deep South" (Mississippi, Louisiana, Arkansas, Tennessee, Alabama, Georgia, North Carolina, South Carolina, and Florida). This Deep-South region is statistically different in CHI scores from the US Census Bureau-defined grouping of states in the South. The mean of CHI scores for the Deep-South region was >1 SD below the mean of CHI scores for all states. In contrast, the CHI score means for each of the other 3 regions were all above the overall mean of CHI scores for all states. Regression analysis showed that living in the Deep-South region is a stronger predictor of poor child health outcomes than other consistently collected and reported variables commonly used to predict children's health. CONCLUSIONS: The findings of this study indicate that region of residence in the United States is statistically related to important measures of children's health and may be among the most powerful predictors of child health outcomes and disparities. This clarification of the poorer health status of children living in the Deep South through spatial analysis is an essential first step for developing a better understanding of variations in the health of children. Similar to early epidemiology work linking geographic boundaries to disease, discovering the mechanisms/pathways/causes by which region influences health outcomes is a critical step in addressing disparities and inequities in child health and one that is an important and fertile area for future research. The reasons for these disparities may be complex and synergistically related to various economic, political, social, cultural, and perhaps even environmental (physical) factors in the region. This research will require the use and development of new approaches and applications of spatial analysis to develop insights into the societal, environmental, and historical determinants of child health that have been neglected in previous child health outcomes and policy research. The public policy implications of the findings in this study are substantial. Few, if any, policies identify these children as a high-risk group on the basis of their region of residence. A better understanding of the depth and breadth of disparities in health, education, and other social outcomes among and within regions of the United States is necessary for the generation of policies that enable policy makers to address and mitigate the factors that influence these disparities. Defining and clarifying the regional boundaries is also necessary to better inform public policy decisions related to resource allocation and the prevention and/or mitigation of the effects of region on child health. The identification of the Deep South as a clearly defined subregion of the Census Bureau's regional definition of the South suggests the need to use more culturally and socially relevant boundaries than the Census Bureau regions when analyzing regional data for policy development. PMID- 16263975 TI - All-terrain vehicle-related nonfatal injuries among young riders: United States, 2001-2003. AB - BACKGROUND: All-terrain vehicles (ATVs) have gained in popularity in recent years, and this rise in use has been accompanied by increases in the number of ATV-related injuries. Because children often lack the physical strength, cognitive abilities, and fine motor skills to operate ATVs properly, their risk for injury is greater. Furthermore, most children ride adult-sized ATVs. OBJECTIVES: To estimate the numbers and rates of ATV-related nonfatal injuries to riders aged < or =15 years who were treated in hospital emergency departments (EDs) in the United States from 2001 through 2003. METHODS: Estimates of ATV related injuries were obtained from the US Consumer Product Safety Commission's National Electronic Injury Surveillance System-All Injury Program. The database is a nationally representative, stratified probability sample of 66 US hospitals with > or =6 beds and a 24-hour ED. ATV-related nonfatal injuries to riders aged < or =15 years who were treated in hospital EDs were examined by age group, gender, primary body part injured, diagnosis, and hospital admission status. RESULTS: From 2001 through 2003, an estimated 108724 children aged < or =15 years were treated in hospital EDs for nonfatal injuries sustained while riding ATVs. The number of ATV-related injuries increased by 25% over the 3-year period. Males aged 11 to 15 years accounted for 52% of all ATV-related ED visits and hospitalizations among young riders. Children aged 0 to 5 years were more likely than the older children to have facial injuries, whereas the older children were more likely to sustain lower trunk and leg or foot injuries. Fractures were the most common diagnosis, accounting for 27% of ED visits and 45% of hospitalizations. CONCLUSIONS: Current legal and regulatory standards have been ineffective in reducing injuries among young ATV riders. Renewed efforts by health care providers to counsel parents about the injury risk to children who ride ATVs and advocate for more stringent state-level minimum age requirements may help reduce the escalating rates of ATV-related injuries among young riders. PMID- 16263974 TI - Cerebral lymphoma in an adenosine deaminase-deficient patient with severe combined immunodeficiency receiving polyethylene glycol-conjugated adenosine deaminase. AB - Polyethylene glycol-conjugated adenosine deaminase (PEG-ADA) provides an alternate therapy to mismatched stem cell transplantation for patients with ADA deficient severe combined immunodeficiency. Although replacement therapy with PEG ADA is effective in preventing infections, immune function does not return to normal, and most patients remain lymphopenic. Information is limited regarding the prognosis of patients on long-term ADA-replacement therapy. Here we present a case of a 10-year-old child who was diagnosed with ADA-severe combined immunodeficiency at 4 weeks of age after contracting pneumonia. Treatment with PEG-ADA was begun, the biochemical markers of ADA deficiency normalized, and his clinical progress was very good without significant infections. At 10 years of age, after presenting with headaches and cranial nerve deficits, he was diagnosed with Epstein-Barr virus-positive malignant brain lymphoma. It did not respond to various regimens of aggressive chemotherapy, and the patient expired 5 months later. We speculate that in this patient the immunologic surveillance by T cells may have been defective with respect to elimination of Epstein-Barr virus infected cells, hence the formation of neoplasm. The possible mechanisms underlying such pathology are reviewed. PMID- 16263976 TI - How do physicians immunize their own children? Differences among pediatricians and nonpediatricians. AB - CONTEXT: Immunization has an essential impact on public health worldwide. Numerous studies have shown the efficacy of different vaccines to protect individuals from various diseases. However, some parents choose not to vaccinate their children for reasons such as, among others, doubts regarding their usefulness, concerns over safety or efficacy, etc. Physicians are known to exert a direct influence on immunization rates by answering questions and clarifying misconceptions. Yet, it is unknown how they immunize their own children. OBJECTIVE: We sought to assess how physicians interested in vaccination issues immunized, or would immunize, their own children. DESIGN, SETTING, AND PARTICIPANTS: An 11-question, Web-based survey with a total of 102 discrete answers was sent to 2070 Swiss physicians in October 2004. All physicians were subscribers to a nonprofit, Web-based expert network (InfoVac, www.infovac.ch) that distributes monthly newsletters and answers question within 2 days on immunization issues. The InfoVac network reaches > 95% of pediatricians in Switzerland but < 20% of general practitioners. All responses were anonymous, and no identifier could be used to trace the participants of the survey. Questions were divided into 2 parts: (1) physicians who were parents were asked which vaccines they gave to their own children and at what age, and (2) all physicians were asked which vaccines they would give to their own child and at what age if they had a newborn child in 2004. Vaccines available in Switzerland at the time of the survey were offered as possible replies, and recommended vaccines were considered as those noted in the Swiss federal immunization schedule issued yearly. One question compared their immunization practice between their own children and their patients. Sociodemographics, qualifying year, membership in different professional groups, and their type of practice were also requested. Statistics. Standard descriptive statistics were used for sociodemographic characteristics. Univariate statistical analyses were performed for each variable to determine its relationship to the dependent variable, being a pediatrician or nonpediatrician. Logistic-regression analysis was used to calculate the adjusted odds ratios (ORs) and 95% confidence intervals (CIs), controlling for any statistically significant demographic variables that might function as confounders (gender, parenthood, workplace, year of diploma, and type of practice). For all statistical tests, differences were considered significant at P < .05. MAIN OUTCOME MEASURE: We performed a comparison of past and projected immunization rates in the children of pediatricians and nonpediatricians. RESULTS: One thousand seventeen valid questionnaires were received (response rate: 49.1%; pediatricians: 53.3%). Nine hundred fifteen physicians (90%) had > or = 1 child. All physicians reported immunizing children in their practice. Pediatricians were more likely to be women and to work in private practice than nonpediatricians but less likely to belong to a self-reported alternative medicine association. Among the nonpediatricians, 317 were general practitioners, 144 were internists, and 95 were other specialists. Ninety-two percent of pediatricians followed the official immunization recommendations for their own children. In contrast, after controlling for gender, workplace, type of practice, and year of diploma, nonpediatricians were more likely not to have immunized their children against measles, mumps, hepatitis B, or Haemophilus influenzae type b. They more frequently postponed diphtheria-tetanus-pertussis (DTP) (OR: 4.5; 95% CI: 2.0-10.19) and measles-mumps-rubella (MMR) vaccination. Although projected immunization rates were higher than effective rates, 10% of nonpediatricians would still not follow the official immunization recommendations in 2004. They would more frequently refrain from using combination vaccines and postpone DTP and MMR immunization to later in life. Several comparisons confirmed the weaker use of the more recently licensed vaccines by nonpediatricians. In addition to vaccines currently recommended in Switzerland, both groups of physicians added hepatitis A, influenza, and varicella vaccines to the vaccination schedule of their own children. Pediatricians were more likely to give pneumococcal (OR: 2.26; 95% CI: 1.004-4.68) and meningococcal C (OR: 2.26; 95% CI: 1.62-3.17) vaccines to their own children. In contrast, they were less likely to give tick-borne encephalitis virus vaccine (OR: 0.65; 95% CI: 0.44 0.95). CONCLUSIONS: Ninety-three percent of the surveyed physicians agree with the current official vaccination recommendations and would apply them to their own children. However, the observation that 5% of nonpediatricians would not use Haemophilus influenzae type b vaccine if they had a child born in 2004 is unexpected and concerning. In contrast, both groups gave additional vaccines than those recommended to their own children. Among physicians in Switzerland interested in immunization, a significant proportion of nonpediatricians decline or delay the immunization of their own children with the recommended MMR- or DTP based combination vaccines, which indicates that clarification of misconceptions such as fear of "immune overload" has not yet reached important targets among health care providers who thus are unlikely to answer parental concerns adequately. PMID- 16263977 TI - The introduction of computerized physician order entry and change management in a tertiary pediatric hospital. AB - OBJECTIVES: The objectives of this review were to document the introduction of computerized physician order entry (CPOE)-centered changes in an academic tertiary care center and to review the CPOE-focused literature. DESIGN: We performed a systematic literature review of CPOE-related articles indexed on Medline, with particular emphasis on pediatric applications. We focused our commentary around the concepts involved in the implementation process at a tertiary pediatric hospital. RESULTS: In 2001, the Children's Hospital of Pittsburgh (CHP) embarked on the process of CPOE design and implementation. We determined that CPOE is a tool for improving pediatric care. The CPOE implementation process is more than a technologic change; it involves an organizational cultural transformation. Although the complete transition to CPOE was little more than 1 year ago, CHP has overcome the typical obstacles of CPOE implementation to begin to realize its many benefits. The early success of CHP was achieved by creating a realistic, positive, work environment, which fostered hospital-wide participation and integration. CONCLUSION: CPOE is an invaluable resource for supporting patient safety in health care settings. The successful implementation of CPOE requires a paradigm shift in hospital policies and processes. PMID- 16263978 TI - Using the internet to provide information prescriptions. AB - INTRODUCTION: An information prescription is the provision of specific information to a patient on how to help manage a health problem. The Internet is being used increasingly as a source for information prescriptions, with clinicians directing patients to specific Web sites. As with any health care intervention, patients' lack of compliance is a barrier to the effectiveness of Web-based information prescriptions (WebIPs). WebIPs cannot be helpful if patients do not review the information prescribed for them. OBJECTIVE: The main objective of this study was to quantify the percentage of families who visit a Web site that was specifically prescribed by their physician. In addition, the use of an e-mail reminder was used to determine if it increases the likelihood that families will visit the prescribed Web site. Finally, barriers to accessing the prescribed Web site were identified. METHODS: Children were eligible if they presented to the pediatric gastroenterology clinic with chronic constipation and/or encopresis and their family had an active e-mail account and access to the Internet in their home. During their clinic visit, physicians instructed families to visit a Web site that provided educational information pertinent to their child's problem. Families were given a form with the Web-site address and a log in identification number. Two days after their clinic visit, half of the families received an e-mail reminding them to visit the Web site. Families were contacted 1 week after their clinic visit to identify barriers to accessing the Web site. RESULTS: Eighty-three families participated in the study. Of the 83 families, 54 (65%) visited the prescribed Web site within 1 week of their clinic visit. Families who received e-mail reminders were significantly more likely to visit the Web site than families who did not receive an e-mail reminder (77% vs 53%). This difference could not be explained by the type or speed of Internet connection or how frequently they accessed the Internet or e-mail. The most common reasons that families cited for not accessing the Web site were "I forgot" and "I didn't have time." Few families cited technical reasons for not accessing the Web site. CONCLUSIONS: Almost two thirds of the families given a WebIP logged on to the prescribed Web site. The probability that families would access the site was increased by 45% with an e-mail reminder. Clearly, e-mail prompts improve compliance to WebIPs. As content and treatment programs continue to proliferate on the Web, it is important to identify barriers and solutions to them to improve overall compliance. PMID- 16263979 TI - Sublingual sugar administration as an alternative to intravenous dextrose administration to correct hypoglycemia among children in the tropics. AB - BACKGROUND: Hypoglycemia is a common determining factor of poor prognosis for children with severe malaria in sub-Saharan Africa. Intravenous dextrose administration is rarely available. Oral mucosal delivery may be an alternative to parenteral administration. A randomized, clinical trial was performed in Burkina Faso among moderately hypoglycemic children, comparing sublingual sugar administration with oral water, oral sugar, and dextrose infusion administrations. METHODS: Sixty-nine children with glucose concentrations of < 0.8 g/L were assigned randomly to 1 of 4 methods of administration, 1 with 3 different doses of sugar, as follows: oral group (OG) (n = 15): 2.5 g of sugar; sublingual group (SG) (n = 27): 2.5 g of sugar under the tongue, with 3 treatment subgroups, ie, 0.1 g/kg, 0.15 g/kg, and 0.2 g/kg; intravenous group (IG) (n = 8): 8 mL of 30% dextrose in a single bolus; water group (n = 11). Eight children received sublingual sugar twice, ie, 0.1 g/kg at baseline and 20 minutes later. Blood glucose concentrations were measured every 20 minutes for 80 minutes. Treatment failures, peak glucose concentrations, times to glucose concentration normalization, and kinetic profiles were evaluated. RESULTS: No treatment failures were observed in the SG and IG, compared with 8 (53%) and 9 (81.8%) failures in the OG and water group, respectively. SG children exhibited glucose kinetic profiles and bioavailabilities (77%, 99%, and 81% in the 3 SG groups) similar to those of IG children. Bioavailabilities were 84% and 38% in the SG and OG, respectively. Children > 7 years of age required repeated sublingual administrations to maintain normoglycemia. CONCLUSIONS: The sublingual administration of sugar proved to be effective among moderately hypoglycemic children. It is a simple and promising method to control hypoglycemia among children in both developing and developed countries. This pediatric practice should be investigated in more detail among children with severe malaria. PMID- 16263980 TI - Nonresponse bias in a follow-up study of 19-year-old adolescents born as preterm infants. AB - OBJECTIVE: To assess the effect of demographic and neonatal risk factors and outcome at the last available assessment on the probability of full responders, postal responders (those who only responded to the mailed questionnaire), or nonresponders in a follow-up study of 19-year-old adolescents who were born as preterm infants. DESIGN: The 19-year follow-up program was part of a large ongoing collaborative study in The Netherlands on the long-term effect of prematurity and dysmaturity on various medical, psychological, and social parameters. In the original cohort, 1338 infants (94%) with a gestational age of < 32 weeks and/or a birth weight of < 1500 g were enrolled. Neonatal mortality was 23% (n = 312), and another 67 children had died between the ages of 28 days and 19 years, leaving 959 survivors (72% of the original cohort) for follow-up at the present assessment. To study the effect of nonresponse, we divided the 959 survivors into 3 groups: full responders (596 [62.1%]), postal responders (109 [11.4%]), and nonresponders (254 [26.5%]). In the 3 groups we compared demographic and neonatal data, as well as outcome at the last available assessment. RESULTS: The odds ratios (ORs) for male versus female for the probabilities of nonresponse and postal response were statistically significant: 2.7 (95% CI: 1.9-3.9) and 1.6 (95% CI: 1.0-2.5), respectively. The same holds for the ORs for non-Dutch versus Dutch and low versus high maternal education for nonresponse: 2.0 (95% CI: 1.3-3.2) and 3.7 (95% CI: 2.0-6.7), respectively. Special education and severe handicap showed a statistically significant influence on nonresponse (OR: 1.6; 95% CI: 1.1-2.4 and OR: 2.6; 95% CI: 1.3-5.2) and postal response (OR: 2.0; 95% CI: 1.2-3.3 and OR: 4.4; 95% CI: 2.0-9.9), respectively. At the age of 19 years, primary school and special education were found significantly more frequent in the postal responders than in the full response group (20% and 21% vs 6% and 12%). The full responders, on the other hand, were higher educated than were the postal responders. CONCLUSIONS: In this follow-up study at the age of 19 years, boys, non-Dutch adolescents, and low maternal education were overrepresented in the nonresponse and postal-response groups. Nonresponse decreased the proportion of infants with adverse outcome in assessed children. To be able to present reliable results for the total group of survivors in long-term follow-up studies, the nonresponse bias needs to be quantified. Therefore, it is evident that more research using statistical methods such as imputation of missing data is needed. PMID- 16263981 TI - Characteristics of pediatric traumatic amputations treated in hospital emergency departments: United States, 1990-2002. AB - OBJECTIVE: To examine the characteristics of children with traumatic amputations and the products associated with these injuries. METHODS: Data regarding amputation injuries to children younger than 18 years treated in US emergency departments from 1990 to 2002 were obtained from the National Electronic Injury Surveillance System. Data included demographics, product involved, and body region injured. RESULTS: An estimated 111,600 children younger than 18 years with amputation injuries were treated in US emergency departments from 1990 to 2002. The average age was 6.18 years (median: 4 years; mode: 1 year old). Males experienced 65.5% of these injuries. Finger amputations accounted for 91.6% of all amputations, ranging from 95.2% among 0- to 2-year-olds to 87.9% among 13- to 17-year-olds. Complete amputations accounted for 70.2% of all amputations among 13- to 17-year-olds compared with 52.6% of amputations among 0- to 2-year-olds. Adolescents also had the highest proportion of amputations resulting in hospital admission (26.6% for 13- to 17-year-olds compared with 11.9% for 0- to 2-year olds). Adolescents had the highest proportion of amputation injuries involving lawn mowers (14.1% for 13- to 17-year-olds compared with 1.4% for 0- to 2-year olds) and the highest proportion of amputations involving tools (29.3% for 13- to 17-year-olds compared with 2.5% for 0- to 2-year-olds). The percentage of amputations involving doors peaked in the youngest age group and decreased as age increased (65.8% of all amputations for 0- to 2-year-olds compared with 14.1% for 13- to 17-year-olds). CONCLUSIONS: To our knowledge, this is the first study to use a nationally representative sample to broadly investigate amputation injuries among children. The majority of traumatic amputation injuries occur to young children, to males, and to fingers and the majority involve doors. Adolescents experience a higher proportion of more serious amputation injuries. Effective interventions exist but are inadequately used to prevent many of these injuries, including door stops and modifications, bicycle-chain and spoke guards, wearing closed-toe footwear while bicycling, a no-mow-in-reverse default feature on ride on lawnmowers with the override switch located behind the mower operator, and a SawStop system on power saws. Use of these technical countermeasures and changes in relevant product standards to promote their implementation and use could lead to a decrease in pediatric traumatic amputations. PMID- 16263982 TI - Association of insulin-like growth factor I and insulin-like growth factor binding protein-3 with intelligence quotient among 8- to 9-year-old children in the Avon Longitudinal Study of Parents and Children. AB - BACKGROUND: Insulin-like growth factor I (IGF-I) is a hormone that mediates the effects of growth hormone and plays a critical role in somatic growth regulation and organ development. It is hypothesized that it also plays a key role in human brain development. Previous studies have investigated the association of low IGF I levels attributable to growth hormone receptor deficiency with intelligence but produced mixed results. We are aware of no studies that investigated the association of IGF-I levels with IQ in population samples of normal children. OBJECTIVES: To investigate the association of circulating levels of IGF-I and its principle binding protein, IGF-binding protein-3 (IGFBP-3), in childhood with subsequent measures of IQ. METHODS: The cohort study was based on data for 547 white singleton boys and girls, members of the Avon Longitudinal Study of Parents and Children, with IGF-I and IGFBP-3 measurements (obtained at a mean age of 8.0 years) and IQ measured with the Wechsler Intelligence Scale for Children (at a mean age of 8.7 years). We also investigated associations with measures of speech and language based on the Wechsler Objective Reading Dimensions test (measured at an age of 7.5 years) and the Wechsler Objective Language Dimensions test (listening comprehension subtest only, measured at an age of 8.7 years). For some children (n = 407), IGF-I (but not IGFBP-3) levels had been measured at approximately 5 years of age in a previous study. Linear regression models were used to investigate associations of the IGF-I system with the measures of cognitive function. RESULTS: Three hundred one boys and 246 girls were included in the sample. IGF-I levels (mean +/- SD) were 142.6 +/- 53.9 ng/mL for boys and 154.4 +/- 51.6 ng/mL for girls. IQ scores (mean +/- SD) were 106.05 +/- 16.6 and 105.27 +/- 15.6 for boys and girls, respectively. IGF-I levels were associated positively with intelligence. For every 100 ng/mL increase in IGF-I, IQ increased by 3.18 points (95% confidence interval [CI]: 0.52 to 5.84 points). These positive associations were seen in relation to the verbal component (coefficient: 4.27; 95% CI: 1.62 to 6.92), rather than the performance component (coefficient: 1.06; 95% CI: -1.67 to 3.78), of IQ. There was no evidence that associations with overall IQ differed between boys and girls. In a data set with complete information on confounders (n = 484), controlling for birth weight (adjusted for gestation), breastfeeding, and BMI slightly strengthened the associations of IGF I levels with IQ. Additionally controlling for maternal education and IGFBP-3 levels attenuated the associations (change in IQ for every 100 ng/mL increase in IGF-I levels: 2.51 points; 95% CI: -0.42 to 5.44 points). The weakening of associations in models controlling for markers of parental socioeconomic position and education could reflect shared influences of parental IGF levels on parents' own educational attainment and their offspring's IGF-I levels. In unadjusted models examining associations of Wechsler Objective Reading Dimensions and Wechsler Objective Language Dimensions test scores with IGF-I levels, there was no strong evidence that performance on either of these tests was associated with circulating IGF-I levels, although positive associations were seen with both measures. Associations between IGF-I levels measured at age 5 and Wechsler Intelligence Scale for Children scores (n = 407) were similar to those for IGF-I levels measured at age 7 to 8. For every 100 ng/mL increase in IGF-I levels at 5 years of age, IQ increased by 2.3 points (95% CI: -0.21 to 4.89 points). CONCLUSIONS: This study provides some preliminary evidence that IGF-I is associated with brain development in childhood. Additional longitudinal research is required to clarify the role of IGF-I in neurodevelopment. Because IGF-I levels are modifiable through diet and other environmental exposures, this may be one pathway through which the childhood environment may influence neurodevelopment. PMID- 16263983 TI - Child deaths resulting from inflicted injuries: household risk factors and perpetrator characteristics. AB - OBJECTIVE: To determine the role of household composition as an independent risk factor for fatal inflicted injuries among young children and describe perpetrator characteristics. DESIGN, SETTING, AND POPULATION: A population-based, case control study of all children < 5 years of age who died in Missouri between January 1, 1992, and December 31, 1999. Missouri Child Fatality Review Program data were analyzed. Cases all involved children with injuries inflicted by a parent or caregiver. Two age-matched controls per case child were selected randomly from children who died of natural causes. MAIN OUTCOME MEASURE: Inflicted-injury death. Household composition of case and control children was compared by using multivariate logistic regression. We hypothesized that children residing in households with adults unrelated to them are at higher risk of inflicted-injury death than children residing in households with 2 biological parents. RESULTS: We identified 149 inflicted-injury deaths in our population during the 8-year study period. Children residing in households with unrelated adults were nearly 50 times as likely to die of inflicted injuries than children residing with 2 biological parents (adjusted odds ratio: 47.6; 95% confidence interval: 10.4-218). Children in households with a single parent and no other adults in residence had no increased risk of inflicted-injury death (adjusted odds ratio: 0.9; 95% confidence interval: 0.6-1.9). Perpetrators were identified in 132 (88.6%) of the cases. The majority of known perpetrators were male (71.2%), and most were the child's father (34.9%) or the boyfriend of the child's mother (24.2%). In households with unrelated adults, most perpetrators (83.9%) were the unrelated adult household member, and only 2 (6.5%) perpetrators were the biological parent of the child. CONCLUSIONS: Young children who reside in households with unrelated adults are at exceptionally high risk for inflicted injury death. Most perpetrators are male, and most are residents of the decedent child's household at the time of injury. PMID- 16263984 TI - Moyamoya syndrome associated with Down syndrome: outcome after surgical revascularization. AB - OBJECTIVES: This study was undertaken to describe the clinical, radiologic, and angiographic features of moyamoya syndrome in a surgical series of children and adults with Down syndrome. We wished to define the features of moyamoya syndrome associated with Down syndrome and to determine the results of surgical revascularization among these patients at early and late follow-up times. METHODS: We reviewed the clinical, radiologic, and angiographic records of all patients with moyamoya syndrome associated with Down syndrome, as a subset of a previously reported, consecutive series of patients who underwent cerebral revascularization surgery with a standardized surgical procedure, pial synangiosis, between January 1, 1985, and June 30, 2004. RESULTS: Of 181 patients with moyamoya syndrome from the initial series who were treated surgically during the study period, 16 patients had Down syndrome (10 female patients and 6 male patients). The average age at onset was 9.3 years (range: 1-29 years); the average age at the time of surgery was 9.8 years (range: 2-29 years). Although the presenting symptoms were transient ischemic attacks for 10 patients and strokes for 6 patients, computed tomographic and/or MRI scans demonstrated bilateral infarctions for 9 patients and unilateral infarctions for 6, with only 1 patient having no imaging evidence of a previous stroke. No cases presented with intracerebral hemorrhage. Preoperative angiography showed the presence of bilateral moyamoya syndrome changes for all patients, including posterior circulation involvement for 8 patients. Surgical treatment included pial synangiosis for all patients, although 1 patient underwent a superficial temporal artery-middle cerebral artery bypass in the contralateral hemisphere. Surgical complications included symptomatic subdural hematomas requiring evacuation, at 48 days and 54 days postoperatively (2 cases), seizures (2 cases), and strokes within 30 days after surgery, at 1 day and 7 days postoperatively (2 cases). Late clinical and radiologic follow-up data (average: 67.6 months; range: 6-146 months) demonstrated no worsening in neurologic status for any patient except for 1 patient who developed a seizure disorder with associated chronic hypocalcemia; she was totally dependent at the 10-year follow-up evaluation, despite no evidence of new infarction since her surgery. There was no clinical or radiologic evidence of new infarction for any patient in late follow-up evaluations. Postoperative angiography, conducted 1 year after surgery for 11 patients, revealed radiologic evidence of good to excellent cerebral revascularization in 85% of the surgically treated hemispheres. Patients were maintained on lifetime aspirin therapy. CONCLUSIONS: The clinical, radiologic, and angiographic features of moyamoya syndrome associated with Down syndrome seem comparable to those of primary moyamoya disease. Cerebral revascularization surgery with the pial synangiosis technique seems to confer long-lasting protection against additional strokes in this patient population. The presence of moyamoya syndrome should be considered in the evaluation of patients with Down syndrome who present with transient ischemic attack-like symptoms. PMID- 16263985 TI - Do baby-friendly hospitals influence breastfeeding duration on a national level? AB - OBJECTIVES: In Switzerland, the Baby-Friendly Hospital Initiative (BFHI) proposed by the United Nations Children's Fund (UNICEF) was introduced in 1993 to promote breastfeeding nationwide. This study reports results of a national study of the prevalence and duration of breastfeeding in 2003 throughout Switzerland and analyzes the influence of compliance with UNICEF guidelines of the hospital where delivery took place on breastfeeding duration. METHODS: Between April and September 2003, a random sample of mothers who had given birth in the past 9 months in Switzerland received a questionnaire on breastfeeding and complementary feeding. Seventy-four percent of the contacted mothers (n = 3032) participated; they completed a 24-hour dietary recall questionnaire and reported the age at first introduction of various foods and drinks. After excluding questionnaires with missing information relevant for the analyses, we analyzed data for 2861 infants 0 to 11 months of age, born in 145 different health facilities. Because it was known whether each child was born in a designated baby-friendly hospital (45 hospitals) or in a health facility in the process of being evaluated for BFHI inclusion (31 facilities), we were able to assess a possible influence of the BFHI on breastfeeding success. For this purpose, we merged individual data with hospital data on compliance with the UNICEF guidelines, from a data source collected on an annual basis for quality monitoring of designated baby-friendly hospitals and health facilities in the evaluation process. Information on actual compliance with the guidelines allowed us to investigate the relationship between breastfeeding outcomes and compliance with UNICEF guidelines. We were also able to compare the breastfeeding results with those for non-baby-friendly health facilities. The comparison was based on median durations of exclusive, full, and any breastfeeding calculated for each group. To allow for other known influencing factors, we calculated adjusted hazard ratios by using Cox regression; we also conducted logistic regression analyses with the 24-hour dietary recall data, to calculate adjusted odds ratios for validation of results from the retrospectively collected data. RESULTS: In 2003, the median duration of any breastfeeding was 31 weeks at the national level, compared with 22 weeks in 1994, and the median duration of full breastfeeding was 17 weeks, compared with 15 weeks in 1994. The proportion of exclusively breastfed infants 0 to 5 months of age was 42% for infants born in baby-friendly hospitals, compared with 34% for infants born elsewhere. Breastfeeding duration for infants born in baby-friendly hospitals, compared with infants born in other hospitals, was longer if the hospital showed good compliance with the UNICEF guidelines (35 weeks vs 29 weeks for any breastfeeding, 20 weeks vs 17 weeks for full breastfeeding, and 12 weeks vs 6 weeks for exclusive breastfeeding). To control for differences in the study population between the different types of health facilities, hazard and odds ratios were calculated as described above, taking into account socioeconomic and medical factors. Although the analysis of the retrospective data showed clearly that the duration of exclusive and full breastfeeding was significantly longer if delivery occurred in a baby-friendly hospital with high compliance with the UNICEF guidelines, whereas this effect was less prominent in other baby-friendly health facilities, this difference was less obvious in the 24-hour recall data. Only for the duration of any breastfeeding could a positive effect be seen if delivery occurred in a baby-friendly hospital with high compliance with the UNICEF guidelines. Known factors involved in the evaluation of baby-friendly hospitals showed the expected influence, on the individual level, on duration of exclusive, full, and any breastfeeding. If a child had been exclusively breastfed in the hospital, the median duration of exclusive, full, and any breastfeeding was considerably longer than the mean for the entire population or for those who had received water-based liquids or supplements in the hospital. A positive effect on breastfeeding duration could be shown for full rooming in, first suckling within 1 hour, breastfeeding on demand, and also the much-debated practice of pacifier use. After controlling for medical problems before, during, and after delivery, type of delivery, well-being of the mother, maternal smoking, maternal BMI, nationality, education, work, and income, all of the factors were still significantly associated with the duration of full, exclusive, or any breastfeeding. CONCLUSIONS: Our results support the hypothesis that the general increase in breastfeeding in Switzerland since 1994 can be interpreted in part as a consequence of an increasing number of baby-friendly health facilities, whose clients breastfeed longer. Nevertheless, several alternative explanations for the longer breastfeeding duration for deliveries that occurred in baby-friendly hospitals can be discussed. In Switzerland, baby-friendly hospitals actively use their certification by UNICEF as a promotional asset. It is thus possible that differences in breastfeeding duration are attributable to the fact that mothers who intend to breastfeed longer would choose to give birth in a baby-friendly hospital and these mothers would be more willing to comply with the recommendations of the UNICEF guidelines. Even if this were the case, however, this selection bias would not explain the differences in breastfeeding duration between designated baby-friendly health facilities with higher compliance with the UNICEF guidelines and those with lower compliance. Especially this last point strongly supports a beneficial effect of the BFHI, because mothers do not know how well hospitals comply with the UNICEF program. The fact that breastfeeding rates have generally improved even in non-baby-friendly health facilities may be indirectly influenced by the BFHI; its publicity and training programs for health professionals have raised public awareness of the benefits of breastfeeding, and the number of professional lactation counselors has increased continuously. Breastfeeding prevalence and duration in Switzerland have improved in the past 10 years. Children born in a baby-friendly health facility are more likely to be breastfed for a longer time, particularly if the hospital shows high compliance with UNICEF guidelines. Therefore, the BFHI should be continued but should be extended to include monitoring for compliance, to promote the full effect of the BFHI. PMID- 16263986 TI - Effect of a low-allergen maternal diet on colic among breastfed infants: a randomized, controlled trial. AB - BACKGROUND: There is controversy regarding whether hypersensitivity to food proteins contributes to colic among breastfed infants. METHODS: A randomized, controlled trial of a low-allergen maternal diet was conducted among exclusively breastfed infants presenting with colic. In the active arm, mothers excluded cow's milk, eggs, peanuts, tree nuts, wheat, soy, and fish from their diet; mothers in the control group continued to consume these foods. Outcomes were assessed after 7 days, as the change in cry/fuss duration over 48 hours, with validated charts. The primary end point was a reduction in cry/fuss duration of > or =25% from baseline. Mothers also assessed the responses to diet with categorical and visual analog scales. RESULTS: Of 107 infants, 90 completed the trial (mean age: 5.7 weeks; range: 2.9-8.6 weeks; 54 male infants). Infants in both groups presented with significant distress (geometric mean: low-allergen group: 690 minutes per 48 hours; control group: 631 minutes per 48 hours). In follow-up assessments on days 8 and 9, there were significantly more responders in the low-allergen group (74% vs 37%), ie, an absolute risk reduction of 37% (95% confidence interval: 18-56%). Cry/fuss duration per 48 hours was reduced by a substantially greater amount in the low-allergen group; the adjusted geometric mean ratio was 0.79 (95% confidence interval: 0.63-0.97), ie, an average reduction of 21% (95% confidence interval: 3-37%). Mothers' subjective assessments of the responses to diet indicated little difference between the groups. CONCLUSION: Exclusion of allergenic foods from the maternal diet was associated with a reduction in distressed behavior among breastfed infants with colic presenting in the first 6 weeks of life. PMID- 16263988 TI - Effectiveness of iron supplementation in a young child with attention deficit/hyperactivity disorder. AB - A 3-year-old child was referred to consultation for hyperactivity, attention deficit, impulsivity, and sleep problems. He met Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition criteria for attention deficit/hyperactivity disorder. At baseline, the Conners' Parent Rating Scale and the Conners' Teacher Rating Scale raw total scores were 30 and 32, respectively. The child had low a serum ferritin level (13 ng/mL). After 8 months of treatment with Tardyferon (ferrous sulfate, 80 mg/day), his serum ferritin increased to 102 ng/mL. Both parents and teachers reported considerable behavioral improvement. The Conners' Parent and Teacher Rating Scale raw total scores decreased to 19 and 13, respectively. This is the first report of the effectiveness of iron supplementation in a young child with attention-deficit/hyperactivity disorder. PMID- 16263987 TI - Review of case-control studies related to breastfeeding and reduced risk of childhood leukemia. AB - OBJECTIVE: To conduct a systematic review to evaluate the evidence for the effect of breastfeeding on the risk of developing childhood leukemia. REVIEW METHODS: We sought studies providing data regarding the association of breastfeeding and occurrence of childhood leukemia. Studies were identified by using Medline, HHS Blueprint for Action on Breastfeeding, US Department of Health and Human Services Office on Women's Health, Cochrane Database of Systematic Reviews, National Centre for Reviews and Dissemination, reference lists, and national experts. Methodologic quality was evaluated for each study by using criteria from the US Preventive Services Task Force and the National Health Service Centre for Reviews and Dissemination. RESULTS: We reviewed 111 citations to identify 32 potentially eligible full-text articles. Of the 10 studies reviewed, only 4 were sufficient to provide at least fair-quality evidence regarding the association between maternal breastfeeding and childhood leukemia. Studies conflicted regarding the protective effect of breastfeeding on childhood leukemia. In the 2 largest and highest quality studies, breastfeeding was associated with a significant risk reduction in one study with longer breastfeeding duration, reflecting greater protection, and a nonsignificant but suggestive difference in the other. Taken together, half of the studies associated breastfeeding with a lower risk of acute lymphocytic leukemia. CONCLUSIONS: There are few high-quality studies that examine the potential for a protective effect of breastfeeding for childhood leukemia. Furthermore, the few studies that exist disagree regarding the association. It is estimated that the United States spends 1.4 billion dollars annually on the treatment of childhood leukemia. Patients, clinicians, and policy makers do not have the data that they need to make decisions regarding this important potential preventive measure. PMID- 16263989 TI - Apnea is not prolonged by acid gastroesophageal reflux in preterm infants. AB - OBJECTIVE: To examine the temporal relationship between apnea and gastroesophageal reflux (GER) and to assess the effect of GER on apnea duration. METHODS: A total of 119 preterm infants underwent 12-hour cardiorespiratory monitoring studies using respiratory inductance plethysmography, heart rate, oxygen saturation (SaO2), and esophageal pH. The studies were scored for GER (pH <4 for > or =5 seconds) and apnea > or =15 seconds or > or =10 seconds that occurred within 30 seconds of GER. Apnea > or =10 seconds was used to assess whether GER would prolong apnea duration. RESULTS: There were 6255 episodes of GER. Only 1% of GER episodes were associated with apnea > or =15 seconds, and there was no difference in apnea rate before, during, or after GER. There was also no difference in rate of apnea > or =10 seconds before versus during GER; however, there was a decrease in apnea rate immediately after GER. The presence of GER during apnea did not prolong apnea duration, and GER had no effect on the lowest SaO2 or heart rate during apnea. CONCLUSION: There is no evidence of a temporal relationship between acid-based GER and apnea in preterm infants. In addition, GER does not prolong apnea duration and does not exacerbate the resultant decrease in heart rate and SaO2. PMID- 16263990 TI - C-reactive protein in the diagnosis, management, and prognosis of neonatal necrotizing enterocolitis. AB - OBJECTIVE: In this prospective, observational study, we determined whether serum C-reactive protein (CRP) correlated with necrotizing enterocolitis (NEC) stages II and III. We hypothesized that serial CRP measurement if used as an adjunct to abdominal radiographs would improve the identification of infants with NEC. METHODS: Serum CRP level was measured every 12 hours for 3 measurements and, when abnormal, once daily. When clinical signs persisted and the initial abdominal radiographs were abnormal, follow-up radiographs were obtained. RESULTS: Of 241 infants who were evaluated for gastrointestinal signs, 11 had ileus or benign pneumatosis intestinalis with persistently normal CRP; gastrointestinal manifestations resolved within 48 hours, antibiotics were discontinued in <48 hours, and feedings were restarted early without complications. Fifty-five infants had NEC stages II and III; all had abnormal CRP regardless of their blood culture results. In infants with stage II NEC, CRP returned to normal at a mean of 9 days except in those who developed complications such as stricture or abscess formation. CONCLUSIONS: In infants with suspected NEC, normal serial CRP values would favor aborted antibiotic therapy and early resumption of feedings. CRP becomes abnormal in both stage II and stage III NEC. In infants with NEC, persistently elevated CRP after initiation of appropriate medical management suggests associated complications, which may require surgical intervention. PMID- 16263991 TI - Heart rate characteristics: novel physiomarkers to predict neonatal infection and death. AB - OBJECTIVE: Monitoring of regulated physiologic processes using physiomarkers such as heart rate variability may be important in the early diagnosis of subacute, potentially catastrophic illness. Early in the course of neonatal sepsis, there are physiomarkers of reduced heart rate variability and transient decelerations similar to fetal distress. The goal of this study was to determine the degree of increased risk for sepsis, urinary tract infection (UTI), and death when these abnormal heart rate characteristics (HRC) were observed. METHODS: We monitored 1022 infants at 2 tertiary care NICUs, 458 of whom were very low birth weight. We calculated an HRC index from validated regression models relating mathematical features of heart rate time series and histograms to episodes of illness. We calculated the risks for adverse events of sepsis, UTI, and death for infants stratified by HRC measurements. RESULTS: Compared with infants with low-risk HRC measurements, infants with high-risk HRC measurements had 5- to 6-fold increased risk for an adverse event in the next day and 3-fold increased risk in the next week. Laboratory tests that were relevant to infection added information to HRC measurements. Infants with both high-risk HRC and abnormal laboratory tests had 6 to 7-fold increased risk for an adverse event in the next day compared with infants who had neither. CONCLUSION: HRC are noninvasively monitored physiomarkers that identify infants in the NICU who are at high risk for sepsis, UTI, and death. PMID- 16263992 TI - Cost and utilization analysis of a pediatric emergency department diversion project. AB - OBJECTIVE: States are struggling to find effective means to decrease Medicaid costs. The objective of this pilot study was to compare emergency department (ED) cost and utilization by members who were enrolled in a pilot program (designed to reduce the use of hospital EDs) with the costs and utilization incurred by a control group. METHODS: A large, private, primary care pediatric practice launched a pilot ED diversion program that provided extended office hours, multiple access locations, and care coordination. Participants in the program were Medicaid recipients who were younger than 18 years. Enrollment in the program was through either patient self-selection or mandatory assignment by the state Medicaid agency. A total of 17,382 children who were enrolled in the enhanced access program (intervention group) and 26,066 Medicaid-eligible children who received services from other local community primary care providers (control group) were included in the study. Children who had chronic health conditions and were receiving Supplemental Security Income benefits were excluded from this analysis. Regression analyses and t tests were applied to analyze the medical claim data that were collected for this project. Three variables were used as dependent variables to measure different aspects of the ED cost and utilization: per member per month cost, per thousand member per month encounter frequency, and per encounter cost. These variables were used to compare the intervention group with the control group for ED claims, as well as for the overall cost of care during the study period. RESULTS: In the 12-month period subsequent to program initiation, the average per member per month cost for ED utilization of the intervention group was 1.36 dollars less than that of the control group. However, there was no significant difference in terms of per-visit cost related to ED utilization. Therefore, the savings seemed to come as a result of a reduction in ED visits, not from reduced cost per visit. On average, children in the intervention group visited the ED approximately 8 fewer times per thousand members per month than the control group, yet there was no significant difference in the overall (ED and non-ED) cost of care between the intervention and control groups. CONCLUSION: Analysis from the first year of this pilot program demonstrates that by providing enhanced, coordinated, primary care access to Medicaid children, the utilization of the ED was significantly lowered among healthy children, whereas the overall cost of care remained the same. PMID- 16263993 TI - Injury prevention advice in top-selling parenting books. AB - OBJECTIVE: Parenting books are a commonly used source of information on how to keep children and adolescents safe from injuries, the leading cause of death and disability for children aged 1 to 18 years. The content and the quality of the messages contained in these books have not been evaluated formally. The objective of this study was to determine the quantity and the quality of injury prevention messages contained in popular parenting books. METHODS: Top-selling parenting books for 2 major booksellers were reviewed to determine the presence and the accuracy of injury prevention messages as compared with those recommended by the American Academy of Pediatrics (AAP) through The Injury Prevention Program (TIPP) for younger children, aged 0 to 12 years, and the American Medical Association (AMA) through its Parent Package for the safety of adolescents. RESULTS: Forty six parenting books were reviewed, including 41 with messages related to younger children and 19 with messages related to adolescents. These books varied widely with regard to the number of injury prevention messages included. Although some books covered the great majority of TIPP messages for parents of young children, others included very few. In the case of books that address safety for adolescents, no book had more than half of the messages recommended by the AMA. Prevention of burns and motor vehicle injury were the most commonly addressed injury prevention topics in the books focused on younger children, whereas gun safety was the most prevalent injury prevention topic in books that focused on adolescents. Books that were authored by physicians addressed more of the recommended topics and messages than books that were written by authors from other professional backgrounds. The quality of messages was good, ie, consistent with the advice given by the AAP and the AMA. In only a few cases, the parenting books gave injury prevention advice that was inconsistent with recommendations. CONCLUSIONS: Overall, books on parenting adolescents are less likely to contain injury prevention messages than those that address younger children. However, the most frequent injury prevention messages for parents of adolescents describe strategies to prevent firearm injury, a leading cause of death for children in this age group. More emphasis should be placed on prevention of motor vehicle injuries, especially as relates to adolescents. Pediatricians and primary care physicians need to be aware of the strengths and weaknesses of parenting manuals in providing adequate guidance related to injury prevention. PMID- 16263994 TI - Prenatal exposure to cigarettes, alcohol, and coffee and the risk for febrile seizures. AB - OBJECTIVE: Febrile seizure is a common type of seizure in childhood, probably caused by both genetic and early environmental factors. Little is known about the effect of environmental factors that operate in prenatal life, although the fetal brain may be particular vulnerable as a result of extensive brain growth and differentiation in this period. We evaluated the association between prenatal exposure to cigarettes, alcohol, and coffee and the risk for febrile seizures in 2 population-based birth cohorts. METHODS: The Aarhus Birth Cohort consisted of 25,196 children of mothers who were scheduled to deliver at Aarhus University Hospital (1989-1996). The Aalborg-Odense cohort consisted of 10,400 children of women who attended antenatal care in Odense or Aalborg (1984-1987). Both cohorts were linked with the Danish National Hospital Register and followed until December 1998 with a complete follow-up. We extracted from medical records additional information on febrile seizures in children in the Aarhus Birth Cohort who were born between 1989 and 1992. RESULTS: We found a slightly increased risk for febrile seizures in children who were exposed to 10 or more cigarettes per day in the Aarhus Birth Cohort, but the corresponding association was weak in the Aalborg-Odense cohort. We found no association between maternal alcohol and coffee consumption and the risk for febrile seizures. The results were similar for simple and complex febrile seizures. CONCLUSIONS: Our data suggest that prenatal exposure to low to moderate levels of alcohol and coffee has no impact on the risk for febrile seizures, whereas a modest smoking effect cannot be ruled out. PMID- 16263995 TI - Parents' perceptions of factors that affect successful diabetes management for their children. AB - OBJECTIVE: To learn which factors parents perceive to be most influential in determining successful type 1 diabetes management. METHODS: A 4-stage mixed qualitative-quantitative method that consists of a series of focus groups, a survey, and in-depth interviews was used to ensure that parents generated, prioritized, and explained their own ideas. In each stage, parents offered a new level of insight into their perception of how children achieve good metabolic control while living as normal a life as possible. The survey responses were divided into statistically different ranks, and the Kruskal-Wallis test was used to compare the results between subgroups. RESULTS: A total of 149 parents participated in the formative qualitative phases, 799 families (66%) responded to the parent-generated survey, and 67 explanatory interviews were conducted. The families who responded to the survey had children of varied ages (mean: 11.9 years; SD: 4.44) and diabetes control (mean hemoglobin A1c: 8.22%; SD: 1.65); 84.1% of respondents were white, 12.3% were black, and 89% were privately insured. The 30 survey items were statistically discriminated into 8 ranks. The items cover a wide range of categories, including concrete ways of achieving better control, families' or children's traits that affect coping ability, actions of the health care team that support versus undermine families' efforts, and the availability of community supports. No clear pattern emerged regarding 1 category that parents perceived to matter most. CONCLUSIONS: Clinicians can affect many of the factors that parents perceive to make a difference in whether they can successfully raise a resilient child in good diabetes control. Future research needs to determine whether health care teams that address the concerns that parents raised in this study are more effective in guiding children to cope well with diabetes, to incorporate healthier lifestyles, and ultimately to achieve better metabolic control. PMID- 16263996 TI - Salivary cortisol and mood and pain profiles during skin-to-skin care for an unselected group of mothers and infants in neonatal intensive care. AB - OBJECTIVES: Mother-infant separation after birth is a well-known source of stress. Parents and preterm infants in neonatal intensive care are separated immediately after birth. Skin-to-skin care is 1 possible method to reduce the separation-dependent stress. The aim of the present study was to investigate how skin-to-skin care influences stress for the mother and the infant in neonatal intensive care. METHODS: Seventeen mother-infant pairs were included at their first and fourth skin-to-skin care. The infants were 25 to 33 weeks' gestational age, with birth weights ranging from 495 to 2590 g. In mothers, salivary cortisol, heart rate, mood scale, and stress measured on a visual analog scale (VAS) were analyzed. In infants, salivary cortisol and heart rate were analyzed, and because pain is one facet of stress, 2 different pain scales were used. RESULTS: In mothers, the skin-to-skin care decreased salivary cortisol (32%), heart rate (7%), and VAS (89%), whereas mood increased (6%). Before the fourth skin-to-skin care, mothers rated less stress on VAS, and salivary cortisol and heart rate improved faster. The infants' cortisol either increased or decreased. Their heart rates and pain scores decreased during skin-to-skin care. CONCLUSIONS: Our results lend additional support to the value of skin-to-skin care in neonatal intensive care. Variable stress responses in preterm infants favor the need for individualized care. The mothers' need for support seem to be more pronounced in the first skin-to-skin session as our results show a higher degree of stress as compared with later skin-to-skin care. PMID- 16263997 TI - Regional variation in rates of low birth weight. AB - OBJECTIVE: Low birth weight (LBW; < 2500 g) is the result of complex and poorly understood interactions between the biological determinants of the mother and the fetus, the parent's socioeconomic status, and medical care. After controlling for these established risk factors, the extent of regional variation in LBW rates remains unknown. This study measures regional variation in LBW rates and identifies regions of neonatal health services with significantly high or low adjusted rates. METHODS: Linking the United States 1998 singleton birth cohort (N = 3.8 million) with county and health care characteristics, we conducted a small area analysis of LBW across 246 regions of neonatal health services. We measured observed rates and then used a multivariable, hierarchical model to estimate adjusted LBW rates by regions. We then stratified these rates by race for the 208 regions with adequate sample size. RESULTS: Observed LBW rates varied across regions from 3.8 to 10.6 per 100 live births (interquartile range: 5.0-6.8 [25th 75th percentile]; median: 5.9). After controlling for known maternal and area risk factors, 67 (27.0%) regions had rates significantly below and 98 (39.8%) regions had rates significantly higher than the national rate of 6.0 per 100 live births. Although black mothers were more likely to give birth to an LBW newborn, regional adjusted rates still varied > 3-fold within both black and nonblack subgroups. CONCLUSIONS: After controlling for known maternal and area risk factors, LBW rates markedly varied across US regions of neonatal health services for both black and nonblack mothers. Additional analyses of these regions may provide opportunities for regional accountability in pregnancy outcomes, LBW research, and targeted improvement interventions, especially in high-risk populations. PMID- 16263998 TI - Prevalence of impaired fasting glucose and its relationship with cardiovascular disease risk factors in US adolescents, 1999-2000. AB - OBJECTIVE: PEDIATRICS (ISSN 0031 4005). Published in the public domain by the American Academy of Pediatrics. Several studies have reported increases in the occurrence of type 2 diabetes in youths. People with prediabetic states such as impaired fasting glucose (IFG) are at increased risk for developing diabetes and cardiovascular disease (CVD). The objective of this study was to examine the prevalence of IFG and its relationship with overweight and CVD risk factors in a nationally representative sample of US adolescents who were aged 12 to 19 years. METHODS: We used data from the 1999-2000 National Health and Nutrition Examination Survey (NHANES). Adolescents who had fasted for 8 hours or more were included in the study (n = 915). IFG was defined as a fasting glucose of 100 to 125 mg/dL. Participants were classified as overweight when their age- and gender specific BMI was > or = 95th percentile and as at-risk for overweight when their BMI was > or = 85th and < 95th percentile. RESULTS: In 1999-2000, the prevalence of IFG in US adolescents was 7.0% and was higher in boys than in girls (10.0% vs 4.0%). Prevalence of IFG was higher in overweight adolescents (17.8%) but was similar in those with normal weight and those who were at risk for overweight (5.4% vs 2.8%). The prevalence of IFG was significantly different across racial/ethnic groups (13.0%, 4.2%, and 7% in Mexican Americans, non-Hispanic black individuals, and non-Hispanic white individuals, respectively). Adolescents with IFG had significantly higher mean hemoglobin A1c, fasting insulin, total and low-density lipoprotein cholesterol, triglycerides, and systolic blood pressure and lower high-density lipoprotein cholesterol than those with normal fasting glucose concentrations. CONCLUSIONS: These data, representing 27 million US adolescents, reveal a very high prevalence of IFG (1 in 10 boys and 1 in 25 girls) among adolescents; the condition affects 1 in every 6 overweight adolescents. Adolescents with IFG have features of insulin resistance and worsened CVD risk factors. Evidence for prevention is still forthcoming in this age group. PMID- 16263999 TI - Helium/oxygen-driven albuterol nebulization in the treatment of children with moderate to severe asthma exacerbations: a randomized, controlled trial. AB - BACKGROUND: Helium and oxygen mixtures (heliox) increase both pulmonary aerosol delivery and gas delivery relative to oxygen. We aimed to compare the effectiveness of a 70%:30% helium/oxygen (heliox)-driven continuous aerosol delivery versus 100% oxygen-driven delivery in the treatment of asthmatic children with moderate to severe exacerbations. METHODS: We enrolled 30 children aged 2 to 18 years who presented to an urban, pediatric emergency department (ED) with moderate to severe asthma as defined by a pulmonary index (PI) score of > or =8. PI scores can range from 0 to 15. In this randomized, controlled, single blind trial conducted in a convenience sample of children, all patients in the trial received an initial nebulized albuterol (5 mg) treatment driven by 100% oxygen and a dose of oral prednisone or prednisolone. Subsequently, patients were randomly assigned to receive continuously nebulized albuterol (15 mg/hour) delivered by either heliox or oxygen using a nonrebreathing face mask. The primary outcome measure was degree of improvement as assessed in blinded video recorded PI scores over 240 minutes (at 30-minute intervals for the first 3 hours) or until ED discharge (if <240 minutes). RESULTS: The mean change in PI score from baseline to 240 minutes or ED discharge was 6.67 for the heliox group compared with 3.33 for the oxygen group. Eleven (73%) patients in the heliox group were discharged from the hospital in <12 hours compared with 5 (33%) patients in the conventional group. CONCLUSION: Continuously nebulized albuterol delivered by heliox was associated with a greater degree of clinical improvement compared with that delivered by oxygen among children with moderate to severe asthma exacerbations. PMID- 16264000 TI - Administration of epinephrine for life-threatening allergic reactions in school settings. AB - OBJECTIVE: Although the potential for life-threatening allergic reactions in children is a significant health concern for schools, there is little information about the circumstances surrounding anaphylactic events that occur in schools. The objectives of this study were to determine the incidence of anaphylaxis in schools, describe the circumstances around anaphylactic events, assess practices that are used to manage students with life-threatening allergies, and identify opportunities for improvement. METHODS: A total of 109 school districts in Massachusetts completed an Epinephrine Administration Form whenever epinephrine was administered at school. Data were collected from September 2001 to August 2003. RESULTS: Forty-eight school districts reported a total of 115 administrations of epinephrine during the 2-year reporting period. In 24% of the cases, the individual was not known to have a life-threatening allergy. Almost one third (31%) of the students who received epinephrine had allergies to multiple substances, and one quarter (25%) had an allergy to peanuts or tree nuts only. Twenty-two (19%) cases occurred outside the school building on the playground, traveling to and from school, or on field trips. The administration of epinephrine most often occurred in the health office by a registered nurse. The average time from onset of symptoms to administration of epinephrine was 10 minutes. In 92% of the cases, the student was transported to a medical facility via the emergency medical system. CONCLUSIONS: Anaphylactic reactions in schools, although not frequent, are not uncommon events. A systematic review of anaphylactic events that required epinephrine administration identified opportunities for improvement in the treatment of students with life-threatening allergies. PMID- 16264001 TI - Epidemiology of pediatric tuberculosis using traditional and molecular techniques: Houston, Texas. AB - OBJECTIVE: To investigate the transmission dynamics of pediatric tuberculosis (TB) by analyzing the clinical characteristics with the molecular profiles of Mycobacterium tuberculosis isolates during a 5-year period. METHODS: A retrospective review of a prospective population-based active surveillance and molecular epidemiology project was conducted in private and public pediatric clinics within Houston and Harris County, Texas. The study population consisted of patients who had pediatric TB diagnosed from October 1, 1995, through September 30, 2000. Cases and potential source cases (PSC) were interviewed using a standardized questionnaire. Available Mycobacterium tuberculosis isolates from cases and PSCs were characterized and compared by IS6110 restriction fragment length polymorphism, spoligotyping, and genetic group assignment. Clinical characteristics were described, and molecular characterizations were compared. Data were analyzed by using EpiInfo 6.02b and SAS 8.2. RESULTS: A total of 220 (92%) of 238 pediatric TB cases were included. Epidemiologic and clinical findings were consistent with previous studies. Molecular profiles from 3 cases did not match the profile of PSC. Four previously unknown PSCs were identified using molecular techniques. Fifty-one (71.8%) of 71 isolates matched at least 1 other Houston Tuberculosis Initiative TB database isolate and were grouped into 33 molecular clusters. Cases were more likely to be clustered when the patients were younger than 5 years, identified a source case, or were US born. CONCLUSIONS: Traditional contact tracing may not always be accurate, and molecular characterization can lead to identification of previously unrecognized source cases. Recent transmission plays a significant role in the transmission of TB to children as evident by the high degree of clustering found in our study population. PMID- 16264002 TI - Medicaid acceptance and availability of timely follow-up for newborns with Medicaid. AB - OBJECTIVE: Decreased physician participation in Medicaid has been shown to affect adversely timeliness of adult acute care and pediatric specialty care, but it is not clear whether this is the case for newborn follow-up. The objectives of this study were to determine whether there is a difference within clinics in the timeliness of follow-up appointments that are given to newborns with Medicaid compared with newborns with private insurance and to determine whether there is a difference between clinics that do and do not accept Medicaid in the timeliness of appointments that are given for newborn follow-up. METHODS: A randomized crossover study was conducted among general pediatric clinics and practices that were identified from the yellow pages and Internet searches of hospitals and health departments in 8 metropolitan areas from September 2003 to March 2004. A simulated parent telephoned clinics to find the earliest available appointment for a 1-day-old infant who needed routine follow-up after discharge that day. Clinics were randomly assigned to receive a first call from a patient with either Medicaid or private insurance; each clinic received the same call at least 3 weeks later with the patient's insurance status reversed. The main outcome measure was whether the appointment was timely (< or =2 days from the day of the call). RESULTS: Of 401 participating clinics, 22% did not accept Medicaid. Among clinics that accepted Medicaid, availability of a timely appointment for a newborn with Medicaid was similar to that for a newborn with private insurance (87% vs 90%, respectively). Appointments that were provided to privately insured newborns were as likely to be timely in clinics that accept Medicaid as in clinics that do not accept Medicaid (89.5% vs 93.4%, respectively). However, providing timely appointments was significantly less likely in clinics that were in high-poverty locations compared with clinics that were not (86.1% vs 92.7%, respectively). CONCLUSIONS: Although newborns with Medicaid did not have access to >20% of clinics because of their insurance, among clinics that did accept Medicaid, timeliness of available follow-up was similar for newborns with Medicaid compared with newborns with private insurance and similar between clinics that did and did not accept Medicaid. However, to the extent that care for newborns with Medicaid is concentrated in clinics in high-poverty areas, some newborns with Medicaid may not be able to receive timely appointments. PMID- 16264004 TI - Association between underinsurance and access to care among children with special health care needs in the United States. AB - OBJECTIVE: To examine the impact of underinsurance on access to care among children with special health care needs (CSHCN) in the United States. METHODS: Interviews were conducted by telephone with the families of 38866 CSHCN who were younger than 18 years using the 2001 National Survey of Children With Special Health Care Needs. The prevalence of underinsurance and its relationship to access to care and family financial problems was examined in this cross-sectional analysis. CSHCN were classified as underinsured when coverage was deemed inadequate to meet the child's needs. RESULTS: An estimated 12.8% of US children experienced a special health care need in 2001. Although 95% of CSHCN had some type of insurance coverage at the time of the interview, 32% were classified as underinsured. Underinsured CSHCN were disproportionately represented in low income families and were significantly more likely than fully insured children to have unmet health needs, and their families were more likely to report difficulty in obtaining specialty referrals, experience financial problems, and report that the child's condition caused family members to reduce or stop work. Underinsured CSHCN seemed to be somewhat better off than CSHCN with no insurance coverage on these measures. CONCLUSIONS: Underinsured CSHCN represent an important and largely hidden underserved population. PMID- 16264003 TI - Insurance gaps among vulnerable children in the United States, 1999-2001. AB - OBJECTIVE: To quantify the number of children who experience gaps in insurance coverage and to determine whether vulnerable subgroups of children experience noteworthy lapses in insurance coverage. METHODS: We analyzed nationally representative data from 24,149 children sampled in the 1999-2001 Medical Expenditure Panel Survey linked to the 1997-1999 National Health Interview Survey. Vulnerable subgroups of children included children with chronic conditions, those from ethnic/racial minorities, and those living in poverty. On the basis of cumulative annual monthly insurance coverage status, each child fell into 1 of 3 groups: continuous coverage, uninsured, or gaps in coverage. Using SAS-callable SUDAAN, we conducted multivariate ordinal logistic regression model to quantify the likelihood of having gaps in coverage for vulnerable subgroups of children. RESULTS: From 1999 to 2001, we found that >9 million American children annually had gaps in coverage and that 5 to 6 million children annually were uninsured for the entire year. Sixty percent of children experienced gaps of at least 4 months, and >40% of all publicly and privately insured children had coverage gaps. After accounting for relevant covariates, children with chronic conditions were just as likely as other children to have gaps in coverage or be uninsured; Hispanic children were most likely to have insurance gaps or be uninsured; and children from poor and near-poor families were 4 to 5 times more likely to have lapsed coverage than children from high-income families. Poverty and maternal education were the strongest factors associated with lapsed coverage. CONCLUSIONS: Unstable health insurance is an underrecognized problem for children, including those with chronic conditions. Because unstable insurance coverage can lead to inadequate health care utilization and poor child health outcomes, strategies to promote stable insurance coverage merit serious consideration. PMID- 16264005 TI - Long-term follow-up of neonatal mitochondrial cytopathies: a study of 57 patients. AB - OBJECTIVES: We sought to determine the long-term clinical and biochemical outcome of newborns with mitochondrial cytopathies (MCs) and to identify possible prognostic factors that may modify the course of these diseases. MATERIAL AND METHODS: Fifty-seven newborns with MCs were identified in a retrospective review (1983-2002). We defined 2 different outcome categories: clinical (neurologic, hepatic, myopathic, and multiorganic) and biochemical (lactate level normalization or initially normal remaining unchanged, decreased but not normalized, and persistently high). We used 2 different statistical approaches: (1) survival studies depending on the initial symptoms and lactate and enzymatic deficiencies using the Kaplan-Meier method; and (2) the same variables compared with different survival age groups and clinical and biochemical outcome categories using the chi2 test. RESULTS: Thirty-three patients died (57.8%), 12 remain alive (21%), and 12 were lost in the follow-up; 6 of them are currently older than 4 years. Most of the patients manifested multiorganic disease (64.8%) and high lactate level (77.1%) over time. Children surviving to 2.5 to 3 years of age were more likely to survive for a long period of time. Initial neurologic and hepatic presentation increased the risk to develop neurologic disease and severe persistent hyperlactacidemia, respectively. Initial severe hyperlactacidemia and combined enzyme deficiencies were significant risk factors for higher mortality and multiorganic disorders. Two patients with exclusively myopathic outcome are alive and cognitively normal at 12 years of life. CONCLUSIONS: Children with neonatal-onset MCs have very high mortality and poor prospects. However, some with life-threatening presentations may gradually improve, giving rise to less severe diseases. Those with exclusively myopathic symptoms have a better prognosis. PMID- 16264006 TI - Development of the metabolic syndrome in black and white adolescent girls: a longitudinal assessment. AB - BACKGROUND: The metabolic syndrome, associated with increased risk of type 2 diabetes mellitus and cardiovascular disease, begins to develop during adolescence. OBJECTIVE: We sought to identify early predictors of the presence of the syndrome at the ages of 18 and 19 years in black and white girls. METHODS: Using longitudinal data on participants from 2 centers in the National Heart, Lung, and Blood Institute Growth and Health Study, a 10-year cohort study, we applied cutoffs from the Adult Treatment Panel III to document changes in the prevalence of abnormal syndrome elements and the syndrome in girls aged 9 and 10 years, when cases were rare, and those aged 18 and 19 years, when prevalence had reached 3%. Longitudinal regression models identified early predictors for the presence of the syndrome. RESULTS: Only 1 girl of each race had > or =3 factors at ages 9 and 10 (0.2%), but 20 black girls (3.5%) and 12 white girls (2.3%) had the syndrome 10 years later. Low high-density lipoprotein cholesterol was prevalent throughout the period in both black and white girls. The prevalence of other variables was low at enrollment but increased during follow-up, except for abnormal triglyceride levels in black girls, which remained low throughout follow up. In multivariate models, early measures of waist circumference and triglyceride level were significant predictors for development of the syndrome. CONCLUSION: The strong association of central adiposity with the development of the metabolic syndrome suggests that early interventions aimed at managing preteen obesity could reduce risk of developing the syndrome. PMID- 16264007 TI - Exposure to movie smoking: its relation to smoking initiation among US adolescents. AB - OBJECTIVE: Regional studies have linked exposure to movie smoking with adolescent smoking. We examined this association in a representative US sample. DESIGN/METHODS: We conducted a random-digit-dial survey of 6522 US adolescents aged 10 to 14 years. Using previously validated methods, we estimated exposure to movie smoking, in 532 recent box-office hits, and examined its relation with adolescents having ever tried smoking a cigarette. RESULTS: The distributions of demographics and census region in the unweighted sample were almost identical to 2000 US Census estimates, confirming representativeness. Overall, 10% of the population had tried smoking. Quartile (Q) of movie smoking exposure was significantly associated with the prevalence of smoking initiation: 0.02 of adolescents in Q1 had tried smoking; 0.06 in Q2; 0.11 in Q3; and 0.22 in Q4. This association did not differ significantly by race/ethnicity or census region. After controlling for sociodemographics, friend/sibling/parent smoking, school performance, personality characteristics, and parenting style, the adjusted odds ratio for having tried smoking were 1.7 (95% confidence interval [CI]: 1.1, 2.7) for Q2, 1.8 (95% CI: 1.2, 2.9) for Q3, and 2.6 (95% CI: 1.7, 4.1) for Q4 compared with adolescents in Q1. The covariate-adjusted attributable fraction was 0.38 (95% CI: 0.20, 0.56), suggesting that exposure to movie smoking is the primary independent risk factor for smoking initiation in US adolescents in this age group. CONCLUSIONS: Smoking in movies is a risk factor for smoking initiation among US adolescents. Limiting exposure of young adolescents to movie smoking could have important public health implications. PMID- 16264008 TI - Influenza vaccine confusion: a call for an alternative evidence-based approach. PMID- 16264009 TI - Nasal cannula use in the preterm infant: oxygen or pressure? PMID- 16264010 TI - Apnea is not prolonged by acid gastroesophageal reflux in preterm infants. PMID- 16264011 TI - Medical futility in the neonatal intensive care unit: hope for a resolution. AB - Contemporary medical practice in the NICU sometimes leads to conflicts between providers and parents in which the parent demands continuation of life-sustaining treatment that the medical team deems medically inappropriate or futile. Such conflicts can be difficult to resolve and trying for all parties. Here we describe a conflict involving a 25-week-gestation, 825-g newborn with multiple intractable medical problems and resolution of the conflict through ethics consultation under provisions of the Texas Advance Directives Act. The process established under Texas law sets conceptual and temporal boundaries around the problem of medical futility and provides a legal safe harbor for physicians who seek to withdraw life-sustaining treatments in the setting of medical futility, allowing resolution of such conflicts in a timely and effective manner. As such, it may provide a model for physicians in other states to follow. PMID- 16264012 TI - Anaphylaxis in a milk-allergic child after ingestion of soy formula cross contaminated with cow's milk protein. AB - In this report we describe a 9-month-old boy with severe persistent asthma and documented cow's milk allergy (presented with eczema and severe systemic reactions) who had an anaphylactic reaction to a soy formula contaminated with cow's milk protein. Quantitative enzyme-linked immunosorbent assay analysis revealed trace quantities of beta-lactoglobulin in the offending soy formula as well as the dry powder. The patient did not demonstrate clinical reactivity to soy protein (negative challenge, tolerated pure soy formula well). Cross contamination of the offending soy formula was presumed to have occurred during food manufacturing. This case demonstrates that trace quantities of cow's milk protein can elicit severe systemic reactions in highly milk-allergic individuals. This infant ingested the equivalent of 0.4 mL of cow's milk from the soy formula as documented by an immunoassay for beta-lactoglobulin. This highlights the ease with which cross-contamination can occur during food processing and reinforces the need for better quality control. PMID- 16264013 TI - Clinical (video) findings and cerebrospinal fluid neurotransmitters in 2 children with severe chronic bilirubin encephalopathy, including a former preterm infant without marked hyperbilirubinemia VIDEO. AB - Chronic bilirubin encephalopathy, characterized clinically by extrapyramidal movement abnormalities, vertical gaze abnormalities, and hearing loss, results from neuronal injury after marked hyperbilirubinemia in term and preterm infants. In premature infants, bilirubin staining of specific brain structures has been described at autopsy after only moderate hyperbilirubinemia, but classic chronic bilirubin encephalopathy without marked hyperbilirubinemia has been reported only rarely. We report a case of a 7-year-old, former 29-weeks' gestation, gravely ill premature infant with a peak bilirubin level of 13.3 mg/dL in the neonatal period. We compare this case with a 12-year-old, former term infant with a peak bilirubin level of 49.4 mg/dL on day 10 of life. Both children have dystonia, athetosis, upward gaze palsy, and sensorineural hearing loss, with MRIs showing characteristic abnormal signal in the globus pallidus. We add previously unreported cerebrospinal fluid neurotransmitter levels that show a mild decrease in the dopamine metabolite homovanillic acid in the former premature infant only. PMID- 16264014 TI - Successful treatment of chronic recurrent multifocal osteomyelitis with tumor necrosis factor-alpha blockage. AB - We describe the case of an 18-year-old girl with chronic recurrent multifocal osteomyelitis (CRMO) over a period of 10 years. She had suffered predominantly from very painful recurrent swelling of her cheeks. Various therapeutic regimens including nonsteroidal antiinflammatory drugs and steroids had shown only a partial or temporary response. Because tumor necrosis factor-alpha-blocking agents have been successfully applied in Crohn's-associated CRMO and the related SAPHO (synovitis, acne, pustulosis, hyperostosis, and osteitis) syndrome, tumor necrosis factor-alpha-blocking therapy with infliximab was initiated. Thereafter, apart from 1 mild episode, no additional recurrences were observed during 21 months of follow-up. Infliximab was well tolerated, and steroids were tapered off. Our observation indicates that infliximab may be an effective therapeutic option in CRMO. PMID- 16264015 TI - Failure to thrive as a manifestation of child neglect. AB - Failure to thrive is a common problem in infancy and childhood. It is most often multifactorial in origin. Inadequate nutrition and disturbed social interactions contribute to poor weight gain, delayed development, and abnormal behavior. The syndrome develops in a significant number of children as a consequence of child neglect. This clinical report is intended to focus the pediatrician on the consideration, evaluation, and management of failure to thrive when child neglect may be present. Child protective services agencies should be notified when the evaluation leads to a suspicion of abuse or neglect. PMID- 16264016 TI - Care coordination in the medical home: integrating health and related systems of care for children with special health care needs. AB - Care coordination is a process that facilitates the linkage of children and their families with appropriate services and resources in a coordinated effort to achieve good health. Care coordination for children with special health care needs often is complicated because there is no single point of entry into the multiple systems of care, and complex criteria frequently determine the availability of funding and services among public and private payers. Economic and sociocultural barriers to coordination of care exist and affect families and health care professionals. In their important role of providing a medical home for all children, primary care physicians have a vital role in the process of care coordination, in concert with the family. PMID- 16264017 TI - Response to the American Academy of Pediatrics report on legalization of marijuana. PMID- 16264018 TI - Prophylactic or therapeutic adenoidectomy? PMID- 16264019 TI - Anemia is an important finding and is likely to respond to iron therapy. PMID- 16264020 TI - Authors incorrectly cited classification of adverse-event codes. PMID- 16264021 TI - Leukemoid reaction and bronchopulmonary dysplasia: a primary inflammatory mechanism? PMID- 16264022 TI - Procalcitonin and vesicoureteral reflux in children with urinary tract infection. PMID- 16264023 TI - Pertussis in adolescents and adults: should we accept the results? PMID- 16264024 TI - Transmission of hepatitis C through swapping body jewelry. PMID- 16264025 TI - Improper stool disposal in neonatal units in developing countries: an unrecognized hazard for nosocomial infection. PMID- 16264027 TI - Infective endocarditis. PMID- 16264028 TI - Thrombocytopenia during childhood: what the pediatrician needs to know. PMID- 16264029 TI - A general pediatric approach to evaluating a short child. PMID- 16264030 TI - Index of suspicion. PMID- 16264031 TI - Clinical trials in idiopathic pulmonary fibrosis: a word of caution concerning choice of outcome measures. PMID- 16264032 TI - Are you pulling my airway? PMID- 16264033 TI - Signalling and transcriptional regulation in inflammatory and immune cells: importance in lung biology and disease. PMID- 16264034 TI - COPD disease progression and airway inflammation: uncoupled by smoking cessation. PMID- 16264035 TI - Value of smear and PCR in bronchoalveolar lavage fluid in culture positive pulmonary tuberculosis. AB - At present, further investigations are needed in patients with suspected pulmonary tuberculosis (TB) and either negative sputum smear or without sputum. The aim of the present study was to analyse the yield of bronchoalveolar lavage fluid (BALF) smear and PCR in patients with confirmed pulmonary TB. Patients with a positive culture for Mycobacterium tuberculosis complex in sputum or BALF were analysed over 5 yrs. In total, 90 out of 230 (39%) patients with culture-positive pulmonary TB had a positive sputum smear, and 120 patients underwent bronchoscopy. BALF smear was positive in 56 (47%), BALF PCR in 93 (78%) patients, and BALF smear and/or PCR was positive in 83%. In total, 71 patients who underwent bronchoscopy and had complete clinical records were further analysed. BALF (smear or Mycobacterium tuberculosis complex-PCR) allowed a rapid diagnosis in 10 (59%) out of 17 patients who had a negative sputum smear, and 49 (91%) out of 54 patients without sputum production. Of these 71 patients, 12 (17%) were only culture positive. Rapid diagnosis of pulmonary TB by smear and/or PCR was made in 190 out of 210 patients (90%) in sputum or BALF. In conclusion, combined use of bronchoalveolar lavage fluid smear and Mycobacterium tuberculosis complex PCR has a good diagnostic yield in patients with sputum smear-negative tuberculosis or without sputum production. PMID- 16264036 TI - Use of continuous positive airway pressure during flexible bronchoscopy in young children. AB - Young children are at increased risk for hypoxaemia and hypercapnia during flexible bronchoscopy due to the small size and increased collapsibility of their airways. Various strategies are used to prevent hypoventilation and to provide oxygen during the procedure. The aim of this study was to assess the impact of continuous positive airway pressure (CPAP) on ventilation during flexible bronchoscopy in infants and young children. Tidal breathing was measured in 16 spontaneously breathing and deeply sedated children, aged 3-25 months, by ultrasound spirometry via an airway endoscopy mask. Measurements were made with the tip of the bronchoscope positioned in the pharynx with no CPAP, and in mid trachea with 0, 5 and 10 cmH2O of CPAP. Transition of the bronchoscope through the vocal cords was associated with significant decreases of tidal volumes (5.0+/ 0.5 versus 3.4+/-0.5 mL.kg(-1)), peak tidal expiratory flows (78+/-12 versus 52+/ 10 mL.s(-1)) and peak tidal inspiratory flows (98+/-15 versus 66+/-12 mL.kg(-1)). CPAP (5-10 cmH2O) induced almost complete reversal of these changes. In conclusion, it is shown here that flexible bronchoscopy in spontaneously breathing young children is associated with significant decreases in tidal volume and respiratory flow. These changes are largely reversible with continuous positive airway pressure. PMID- 16264037 TI - Necrotising sarcoid granulomatosis: clinical, functional, endoscopical and radiographical evaluations. AB - Necrotising sarcoid granulomatosis (NSG) is a rare disease diagnosed on the basis of pathological features. The present study reports the characteristics of 14 cases of NSG. The mean age at the appearance of first symptoms was 37 yrs and the mean delay between first symptoms and diagnosis was 1 yr. Extrarespiratory symptoms were more common (12 out of 14) than respiratory symptoms (eight out of 14). Seven patients had inflammatory syndrome. Bronchoalveolar lavage was performed in eight patients and found to be normal in three cases. Respiratory function was normal in 13 patients, but carbon monoxide diffusing capacity was slightly decreased in eight of the 11 patients tested. A computed tomography scan showed a solitary nodule in four out of 14 cases, bilateral nodules in three and infiltrates in seven. One patient died from neurological complications despite treatment with corticosteroids and immunosuppressive drugs. Two cases of relapse were observed in patients initially treated with corticosteroids, and there were two cases of relapse after surgery. No relapse occurred in the five untreated patients. During the follow-up, lung cancer was detected at 26 months and 8 yrs, respectively, after NSG diagnosis in two patients. In conclusion, no one treatment is associated with a better outcome than the others, although lung biopsy might be necessary in case of isolated nodule or cavitation. Greater vigilance is required during the follow-up. PMID- 16264038 TI - Granulocytosis and increased adhesion molecules after resistive loading of the diaphragm. AB - Upregulation of endothelial cell adhesion molecules, followed by an influx of granulocytes and macrophages, can contribute to exertion-induced skeletal muscle injury. The purpose of this study was to quantify circulating leukocyte subsets, diaphragm injury and infiltrating leukocyte subsets, and surface expression of vascular cell adhesion molecule (VCAM)-1 and intracellular adhesion molecule (ICAM)-1 in the diaphragm after inspiratory resistive loading (IRL). Eight New Zealand white rabbits underwent 1.5 h of IRL and seven control rabbits underwent a sham procedure. Blood samples, taken at baseline and 2, 6, 12, 24, 48 and 72 h after the onset of IRL or sham, showed that band cell counts had increased at 6 h post-IRL. Point counting of haematoxylin and eosin-stained cross-sections, sampled at 72 h post-IRL, showed greater injury in diaphragms from IRL rabbits compared with controls. Immunohistochemical processing showed increased expression of ICAM-1 and VCAM-1, and higher granulocyte and macrophage counts in IRL diaphragms than control diaphragms. Macrophages were the predominant inflammatory cells. Increased intracellular adhesion molecule-1 and vascular cell adhesion molecule-1 expression, and infiltration of granulocytes and macrophages may contribute to inspiratory resistive loading-induced diaphragm injury. PMID- 16264039 TI - dsRNA enhances eotaxin-3 production through interleukin-4 receptor upregulation in airway epithelial cells. AB - The exacerbation of asthma during viral infections is mainly explained by neutrophils infiltrating into the airways. However, enhanced functions of eosinophils are also observed. The aim of this study was to reveal the mechanism of how eosinophils are activated during and after viral infection of the airways, using a model of viral infection. A synthetic double-stranded RNA, poly inosinic cytidyric acid (poly(IC)), was transfected to a human airway epithelial cell line (BEAS-2B) and the primary bronchial epithelial cells, to mimic a viral infection. The production of chemokines from the cells was investigated. The transfection of poly(IC), alone, marginally affected the eotaxin-3 production of the cells. However, the transfection of poly(IC) prior to interleukin (IL)-4 stimulation enhanced eotaxin-3 production. Poly(IC) transfection increased mRNA and protein expressions of IL-4 receptor (R)alpha and IL-2Rgamma, components of the IL-4R. In BEAS-2B cells, IL-4-mediated phosphorylation of signal transducer and activator of transcription six was enhanced in poly(IC) transfected cells. This was reversed by the addition of anti-IL-4Ralpha antibody, suggesting the role of an increased number of IL-4 receptors in enhanced IL-4-induced eotaxin-3 production. Poly(IC)-induced upregulation of IL-4Ralpha was inhibited by treatment with cycloheximide or dexamethasone. In conclusion, these results suggest that viral airway infection may enhance interleukin-4-induced eotaxin-3 production through upregulation of the interleukin-4 receptor in airway epithelial cells. PMID- 16264040 TI - Airway narrowing in porcine bronchi with and without lung parenchyma. AB - During bronchoconstriction elastic after-loads arise due to distortion of lung parenchyma by the narrowing airway. In the present study, the functional effect of parenchymal elastic after-load on airway narrowing was determined. Airway narrowing was measured in vivo over a range of transpulmonary pressures and compared with in vitro narrowing measured at corresponding transmural pressures. Bronchi were generation 10 with internal diameters of approximately 4 mm. In vivo luminal narrowing was measured by videobronchoscopy in anaesthetised and ventilated pigs. In vitro luminal narrowing was measured by videoendoscopy in isolated bronchial segments. Airways were activated by maximum vagal nerve stimulation and maximum electrical field stimulation in vivo and in vitro, respectively. At 5 cmH2O, stimulation produced a 35.9+/-3.2% (n = 6) and a 36.5+/ 2.4% (n = 11) decrease in lumen diameter in vivo and in vitro, respectively. At 30 cmH2O, luminal narrowing fell to 23.7+/-2.0% in vivo and 23.4+/-2.5% in vitro. There was no difference between luminal narrowing in vivo and in vitro at any pressure. In conclusion, these findings suggest that in mid-sized, cartilaginous bronchi, parenchymal elastic after-loads do not restrict airway narrowing. PMID- 16264041 TI - Risk factors of frequent exacerbations in difficult-to-treat asthma. AB - Recurrent exacerbations are a major cause of morbidity and medical expenditure in patients with asthma. Various exogenous and endogenous factors are thought to influence the level of asthma control, but systematical data on the involvement of these factors in the recurrence of asthma exacerbations are scarce. In this study, 13 clinical and environmental factors potentially associated with recurrent exacerbations were investigated in 136 patients with difficult-to-treat asthma. Patients with more than three severe exacerbations (n = 39) in the previous year were compared with those with only one exacerbation per year (n = 24). A systematic diagnostic protocol was used to assess 13 potential risk factors. Factors significantly associated with frequent exacerbations included: severe nasal sinus disease (adjusted odds ratio (OR) 3.7); gastro-oesophageal reflux (OR 4.9); recurrent respiratory infections (OR 6.9); psychological dysfunctioning (OR 10.8); and obstructive sleep apnoea (OR 3.4). Severe chronic sinus disease and psychological dysfunctioning were the only independently associated factors (adjusted OR 5.5 and 11.7, respectively). All patients with frequent exacerbations exhibited at least one of these five factors, whilst 52% showed three or more factors. In conclusion, the results show that recurrent exacerbations in asthma are associated with specific co-morbid factors that are easy to detect and that are treatable. Therapeutic interventions aimed at correcting these factors are likely to reduce morbidity and medical expenditure in these patients. PMID- 16264042 TI - Budesonide/formoterol maintenance and reliever therapy: an effective asthma treatment option? AB - This 12-month dose-titration study assessed the effectiveness of budesonide/formoterol for maintenance plus relief with a control group using salmeterol/fluticasone for maintenance plus salbutamol for relief. Adolescents and adults (n = 2,143; mean forced expiratory volume in one second (FEV1) 73% predicted; mean inhaled corticosteroid (ICS) 884 microg.day(-1)) were randomised to budesonide/formoterol 160/4.5 microg two inhalations b.i.d. plus additional inhalations as needed, or salmeterol/fluticasone 50/250 microg b.i.d. plus salbutamol as needed. Treatment was prescribed open label; after 4 weeks, physicians could titrate maintenance doses in accordance with normal clinical practice. Maintenance plus as-needed budesonide/formoterol prolonged the time to first severe exacerbation versus salmeterol/fluticasone (25% risk reduction). The total number of severe exacerbations was significantly reduced in the budesonide/formoterol group (255 versus 329). Both regimens provided sustained improvements in symptoms, as-needed use, quality of life and FEV1, with differences in favour of the budesonide/formoterol group for as-needed use (0.58 versus 0.93 inhalations.day(-1)) and FEV1 (post-beta2-agonist values). Mean ICS dose during treatment was similar in both groups (653 microg budesonide.day(-1) (maintenance plus as-needed) versus 583 microg fluticasone.day(-1)). The simplified strategy using budesonide/formoterol for maintenance and reliever therapy is feasible, safe and at least as effective as salmeterol/fluticasone plus salbutamol. PMID- 16264043 TI - Perceptions of asthma among physicians: an exploratory study with the ISAAC video. AB - The current study examined the perception and interpretation of asthma symptoms among practitioners using standardised audiovisual presentations of asthma. Two groups of practitioners (n = 70) in Chennai, India, were shown the International Study of Asthma and Allergies in Childhood (ISAAC) video questionnaire and asked to describe the symptoms and signs they observed and to identify possible diagnoses for each presentation. The number of practitioners who correctly described the principal symptom(s) of asthma depicted in the five video sequences ranged from 26.1% for scene 5 (wheezing and dyspnoea), to 94.2% for scene 4 (nocturnal cough). The number who identified asthma as a possible cause of the presentations ranged from 17.4% for scene 4, to 67.1% for scene 2 (wheeze after exercise). Practitioners with postgraduate medical education were significantly more likely to identify asthma as a possible cause of the presentations, as were practitioners with postgraduate training in respiratory diseases. In conclusion, the perceptions of asthma and asthma symptoms among many physicians in Chennai, India, do not match the presentations of asthma depicted in the International Study of Asthma and Allergies in Childhood (ISAAC) video. These differences may be limiting the diagnosis and apparent prevalence of asthma, and suggest the need for additional attention to asthma in the education and training of practitioners in India. PMID- 16264044 TI - Effect of 1-year smoking cessation on airway inflammation in COPD and asymptomatic smokers. AB - Smoking cessation is the only treatment in patients with chronic obstructive pulmonary disease (COPD) effective in slowing down disease progression. Its effect on airway inflammation in COPD is unknown, although cross-sectional studies suggest ongoing inflammation in ex-smokers. In order to elucidate the effect of smoking cessation on airway inflammation, 28 smokers with COPD (mean age: 55 yrs; forced expiratory volume in one second: 71% predicted) and 25 asymptomatic smokers with normal lung function (aged 50 yrs) were included in a 1 yr smoking cessation programme. Effects of smoking cessation on airway inflammation were investigated in bronchial biopsies (baseline, 12 months) and sputum samples (baseline, 2, 6 and 12 months). In the 12 candidates with COPD who successfully ceased smoking, airway inflammation persisted in bronchial biopsies, while the number of sputum neutrophils, lymphocytes, interleukin (IL)-8 and eosinophilic-cationic-protein levels significantly increased at 12 months. In the 16 asymptomatic smokers who successfully quitted, inflammation significantly reduced (i.e. number of sputum macrophages, percentage of eosinophils and IL-8 levels) or did not change. The current authors suggest that the observed persistent airway inflammation in patients with chronic obstructive pulmonary disease is related to repair of tissue damage in the airways. It remains to be elucidated whether this reflects a beneficial or detrimental effect. PMID- 16264045 TI - Epidemiological relationships between the common cold and exacerbation frequency in COPD. AB - Higher exacerbation incidence rates in chronic obstructive pulmonary disease (COPD) are associated with more rapid decline in lung function and poorer quality of life, yet the mechanisms determining susceptibility to exacerbation remain ill defined. The same viruses responsible for common colds are frequently isolated during exacerbations. The current authors hypothesised that exacerbation frequency may be associated with an increased frequency of colds, and investigated whether increased exacerbation frequency was associated with increased acquisition of colds, or a greater likelihood of exacerbation once a cold has been acquired. A total of 150 patients with COPD completed diary cards recording peak expiratory flow, and respiratory and coryzal symptoms for a median 1,047 days. Annual cold and exacerbation incidence rates (cold and exacerbation frequency) were calculated, and the relationships between these variables were investigated. This analysis is based on 1,005 colds and 1,493 exacerbations. Frequent exacerbators (i.e. those whose exacerbation frequency was greater than the median) experienced significantly more colds than infrequent exacerbators (1.73 versus 0.94.yr(-1)). The likelihood of exacerbation during a cold was unaffected by exacerbation frequency. Patients experiencing frequent colds had a significantly higher exposure to cigarette smoke (46 versus 33 pack-yrs). Exacerbation frequency in chronic obstructive pulmonary disease is associated with an increased frequency of acquiring the common cold, rather than an increased propensity to exacerbation once a cold has been acquired. PMID- 16264046 TI - Self-management reduces both short- and long-term hospitalisation in COPD. AB - The aim of the present study was to assess the long-term impact on hospitalisation of a self-management programme for chronic obstructive pulmonary disease (COPD) patients. A multicentre, randomised clinical trial was carried out involving 191 COPD patients from seven hospitals. Patients who had one or more hospitalisations in the year preceding study enrolment were assigned to a self management programme "Living Well with COPD(TM)" or to standard care. Hospitalisations from all causes were the primary outcome and were documented from the provincial hospitalisation database; emergency visits were recorded from the provincial health insurance database. Most patients were elderly, not highly educated, had advanced COPD (reflected by a mean forced expiratory volume in one second of 1 L), and almost half reported a dyspnoea score of 5/5 (modified Medical Research Council). At 2 years, there was a statistically significant and clinically relevant reduction in all-cause hospitalisations of 26.9% and in all cause emergency visits of 21.1% in the intervention group as compared to the standard-care group. After adjustment for the self-management intervention effect, the predictive factors for reduced hospitalisations included younger age, sex (female), higher education, increased health status and exercise capacity. In conclusion, in this study, patients with chronic obstructive pulmonary disease who received educational intervention with supervision and support based on disease-specific self-management maintained a significant reduction in hospitalisations after a 2-year period. PMID- 16264047 TI - Goal-oriented treatment and combination therapy for pulmonary arterial hypertension. AB - Combination therapy may improve outcome in patients with severe pulmonary arterial hypertension (PAH). PAH patients were treated according to a goal oriented therapeutic strategy. Patients who did not reach the treatment goals with monotherapy received combination treatment according to a predefined strategy, including bosentan, sildenafil and inhaled iloprost. Intravenous iloprost and lung transplantation were reserved for treatment failures. End points were overall survival, transplantation-free survival, and survival free from transplantation and intravenous prostanoid treatment. Between January 2002 and December 2004, 123 consecutive patients with PAH were treated according to the novel approach. Survival at 1, 2 and 3 yrs was 93.0, 83.1 and 79.9%, respectively, which was significantly better than the survival of a historical control group, as well as the expected survival. Compared to the historical control group, the use of combination treatment also significantly improved the combined end point of death, lung transplantation and need for intravenous iloprost treatment. In conclusion, a therapeutic approach utilising combinations of bosentan, sildenafil and inhaled iloprost in conjunction with a goal-oriented treatment strategy provides acceptable long-term results in patients with severe pulmonary arterial hypertension, and reduces the need for intravenous prostaglandin treatment and lung transplantation. PMID- 16264048 TI - Efficacy of thrombolytic agents in the treatment of pulmonary embolism. AB - Recent guidelines recommend bolus-dose alteplase for treating massive pulmonary embolism (PE). However, the safest and most effective treatment is as yet unknown. In the present study, a meta-analysis of published studies of alteplase infusion, bolus-dose alteplase and streptokinase was performed. The outcome measures were as follows: objective assessment of thrombolysis; all-cause mortality; deaths due to initial PE, major bleeding episodes and recurrent PE; and morbidity. In total, 26 studies were identified; however, only two comparative studies of alteplase infusion versus either bolus-dose alteplase or streptokinase were found. Meta-analysis revealed no significant difference between the three regimens, but was compromised by a paucity of data. Crude analysis of summated data on thrombolytic efficacy from all studies revealed that alteplase infusion was more effective than bolus-dose alteplase (relative risk (RR): 1.95; 95% confidence interval (CI): 1.19-3.2), whereas streptokinase was more effective than alteplase infusion (RR: 1.27; 95% CI: 1.09-1.47). Alteplase infusion had a lower mortality due to the initial PE than both bolus-dose alteplase and streptokinase (RR: 0.16; 95% CI: 0.05-0.59 and RR: 0.13; 95% CI: 0.04-0.46, respectively). In conclusion, this evidence suggests that the three thrombolytic agents may vary in efficacy. However, large-scale randomised controlled trials are needed to confirm these results. PMID- 16264049 TI - Nonmalignant pleural lesions due to environmental exposure to asbestos: a field based, cross-sectional study. AB - The aim of the present study was to investigate the frequencies of asbestos related benign pleural diseases in villagers environmentally exposed to asbestos and the factors affecting these frequencies. A field-based, cross-sectional study was designed. In total, 991 villagers from 10 villages, randomly chosen from 67 villages with known use of asbestos-containing white soil in central Anatolia (Turkey), were investigated. The type of asbestos in the white soil was determined for each village, as well as air-borne fibre concentrations. The villagers were investigated with small-size chest roentgenograms and epidemiological features were recorded. The air-borne fibre levels were generally low. Despite this, pleural plaques were found in 14.4% of the villagers, 10.4% had diffuse pleural fibrosis and 0.4% asbestosis. The significant variables for plaques were age, sex, type of asbestos fibre, e.g. actinolite and tremolite, and exposure duration, while there was a negative relationship with cumulative exposure. The variables affecting diffuse pleural fibrosis were age, exposure duration and cumulative exposure. In conclusion, villagers exposed to environmental asbestos have high frequencies of pleural plaques and diffuse pleural fibrosis, similar to occupationally exposed asbestos cohorts. Different types of asbestos appear to result in different frequencies of pleural lesions. PMID- 16264050 TI - A 20-year follow-up study on chronic respiratory effects of exposure to cotton dust. AB - In order to evaluate chronic effects of long-term exposure to cotton dust on respiratory health, and the role of dust and endotoxin, longitudinal changes in lung function and respiratory symptoms were observed prospectively from 1981 to 2001 in 447 cotton textile workers, along with 472 silk textile controls. The results from five surveys conducted over the 20-yr period are reported, including standardised questionnaires, pre- and post-shift spirometric measurements, work area inhalable dust sample collections and airborne Gram-bacterial endotoxin analysis. Cotton workers had more persistent respiratory symptoms and greater annual declines in forced expiratory volume in one second (FEV1) and forced vital capacity as compared with silk workers. After exposure cessation, in the final 5 yr period, the rate of FEV1 decline tended to slow in nonsmoking males, but not in nonsmoking females. Workers who reported byssinotic symptoms more persistently suffered greater declines in FEV1. Chronic loss in lung function was more strongly associated with exposure to endotoxin than to dust. In conclusion, the current study suggests that long-term exposure to cotton dust, in which airborne endotoxin appears to play an important role, results in substantial adverse chronic respiratory effects. PMID- 16264051 TI - The effects of Aspergillus fumigatus challenge on exhaled and nasal NO levels. AB - Several studies have previously shown that exposure to indoor air microbes from moisture-damaged buildings can cause adverse health effects. Aspergillus fumigatus is one of the best-documented moulds causing health problems to those exposed. In this study, inhalation of a commercial A. fumigatus solution was assessed, to establish if it would have effects on fractional exhaled (FeNO) and nasal (FnNO) nitric oxide levels and on lung function. The results were compared with placebo challenge. A total of 28 subjects were divided into three study groups: group 1 had been exposed to occupational mould; group 2 consisted of atopic subjects; and group 3 was a control group. Some 3 h after A. fumigatus challenge, there was a considerable increase in FeNO, and a significant difference was observed between the A. fumigatus and placebo inhalations. The difference was seen in all study groups. No such differences were found in the levels of FnNO or nitrite in nasal lavage fluid. Subjects reported significantly more frequent respiratory tract symptoms after the A. fumigatus inhalation compared with placebo challenge. In conclusion, it was shown here that inhalation challenge of Aspergillus fumigatus elevated fractional exhaled nitric oxide levels. An increase in fractional exhaled nitric oxide may serve as an indicator of respiratory inflammation of acute mould exposure. PMID- 16264052 TI - The Chrispin-Norman score in cystic fibrosis: doing away with the lateral view. AB - The Chrispin-Norman chest radiograph scoring (CNS) system is widely used to assess respiratory disease progression in cystic fibrosis (CF). Frontal and lateral chest radiographs were performed. The present authors developed a modified CNS, which obviates the need for the lateral film. This study compares the original and the current authors' modified scoring system. A total of 50 chest radiographs from CF children, taken between August and December 2003, were scored according to the original and modified CNS. Two observers scored all 50 chest radiographs, scoring in random order the frontal radiographs, and separately the frontal and lateral radiographs together. There was no evidence of a difference between the methods for either observer, using the Bland and Altman 95% limits of agreement as follows: observer 1 (-2.0-1.9), and observer 2 (-1.77 2.2). No evidence of a difference between the observers for either method was found, comparing the 95% limits of agreement (-5.5-5.7) with the modified CNS ( 5.6-6.4). In conclusion, in terms of the final score, good agreement was found between the use of the original and modified Chrispin-Norman score. In addition, low inter-observer variability was shown for both methods. The use of the modified Chrispin-Norman chest radiograph scoring system to stage disease severity in cystic fibrosis removes the need for a lateral chest radiograph. PMID- 16264053 TI - Driving simulator and neuropsychological [corrected] testing in OSAS before and under CPAP therapy. AB - Patients with obstructive sleep apnoea syndrome (OSAS) have an increased car accident rate. Investigations on accident frequency are based on case history, insurance reports and driving simulator studies. The present study combines neuropsychological testing of different attention aspects engaged in driving a car and driving simulation to evaluate a suitable instrument for assessing therapeutic effects of continuous positive airway pressure (CPAP). Driving simulator investigation and neuropsychological testing of alertness, vigilance and divided attention were performed in 31 patients with polysomnographically confirmed OSAS (apnoea-hypopnoea index 24.8+/-21.5.h(-1)) before, and 2 and 42 days after initiation of CPAP. Divided attention and alertness improved significantly during CPAP, whereas vigilance remained unchanged. However, accident frequency (OSAS before therapy: 2.7+/-2.0; 2 days after CPAP: 1.5+/-1.4; 42 days after CPAP: 0.9+/-1.3) and frequency of concentration faults (OSAS before therapy: 12.4+/-5.1; 2 days after CPAP: 6.5+/-3.9; 42 days after CPAP: 4.9+/-3.3) decreased in the simulated driving situation after 2 and 42 days of therapy. There was no relation between accident frequency, concentration faults and daytime sleepiness, as measured by the Epworth Sleepiness Scale, and polysomnographic or neuropsychological findings, respectively. In conclusion, the present results suggest that driving simulation is a possible benchmark parameter of driving performance in obstructive sleep apnoea syndrome patients. PMID- 16264054 TI - Effect of airway clearance techniques on the efficacy of the sputum induction procedure. AB - Sputum induction is used in the early identification of tuberculosis (TB) and pneumocystis infections of the lung. Although manual physiotherapy techniques to clear the airways are often incorporated in the sputum induction procedure, their efficacy in this setting is unknown. This randomised, crossover trial enrolled adults referred for sputum induction for suspected TB and pneumocystis infections of the lung. All participants underwent two sputum induction procedures, inhaling 3% saline via ultrasonic nebuliser. During one randomly allocated procedure, airway clearance techniques (chest wall percussion, vibration, huffing) were incorporated. In total, 59 participants completed the trial. The airway clearance techniques had no significant effect on how the test was tolerated, the volume expectorated or the quality of the sample obtained (assessed by the presence of alveolar macrophages). The techniques did not significantly affect how often the test identified a suspected organism, nor the sensitivity or specificity of sputum induction. In conclusion, the study was unable to demonstrate any effect of airway clearance techniques on the sputum induction procedure. The results provide some justification for not including airway clearance techniques as part of the sputum induction procedure. PMID- 16264055 TI - alpha1-Antitrypsin deficiency and lung disease: risk modification by occupational and environmental inhalants. AB - Chronic obstructive pulmonary disease (COPD) is a prevalent and preventable disease associated with high morbidity and mortality. Severe and intermediate alpha1-antitrypsin (AAT) deficiency (serum levels <11 and 11-20 micromol.L(-1), respectively) increase the risk of COPD in active smokers. However, little is known about the interaction of severe and intermediate AAT deficiency with modifiable COPD risk factors other than active smoking. In this study, a MEDLINE search was carried out for studies investigating the combined effect of environmental inhalants (occupation and passive smoking) and AAT deficiency in the lung. A total of 18 studies using established methods for the assessment of AAT deficiency were included in this review. Occupational exposures and passive smoking affected lung function decline or prevalence of respiratory symptoms in four out of five studies investigating subjects with severe AAT deficiency, and in eight out of 13 studies with a focus on intermediate AAT deficiency. While study designs mostly prohibited formal assessment of effect modification, an interaction between intermediate AAT deficiency and passive smoking was identified in two studies with children. Additional study limitations included small sample size, poor adjustment for confounding and misclassification of environmental exposure as well as AAT activity. In conclusion, population-based epidemiological studies with associated biobanks are needed to identify gene environment interactions and population subgroups susceptible to alpha1 antitrypsin deficiency. PMID- 16264056 TI - Regulatory T-lymphocytes in asthma. AB - T-helper cell type (Th)2 lymphocytes play an important role in the initiation, progression and persistence of allergic diseases, including asthma. However, little is known about immunoregulatory mechanisms that determine susceptibility to, severity of, or persistence of asthma. The concept of a disturbed Th1/Th2 balance, although having furthered the present understanding of immunoregulation in asthma, has recently been named a "procrustean paradigm", because of its failure to adequately explain many (pre)clinical observations. In recent years, the general knowledge regarding the regulation of infectious, autoimmune diseases, asthma and allergen immunotherapy by T-regulatory (Treg) cells, has rapidly increased. Many different Treg subsets have been described, including CD8+ Treg cells, natural killer (NK) cells and several different CD4+ Treg cell subsets. In this review, the authors will focus on two major and well-described CD4+ Treg cell subsets. These consist of naturally occurring CD25+ Treg cells and adaptive Treg cells that are postulated to prevent immune responses against self antigens and adaptive immune responses, respectively. The adaptive T-regulatory cells are further subdivided into T-regulatory cells type 1 and T-helper cell type 3 that mediate suppression exclusively via the cytokines interleukin-10 and transforming growth factor-beta, respectively. PMID- 16264057 TI - The role of post-transcriptional regulation in chemokine gene expression in inflammation and allergy. AB - The aim of this review is to discuss recent advances in the understanding of the regulation of chemokine expression occurring during chronic inflammatory conditions, such as allergic diseases. The focus will be on current data, which suggest that post-transcriptional regulation plays a larger role in chemokine gene regulation than previously recognised. In particular, a growing body of data indicates that mechanisms controlling mRNA stability may be relevant in determining, or maintaining, the increased levels of chemokine gene expression in this context. Such regulatory pathways may be important targets of novel anti inflammatory strategies. PMID- 16264058 TI - Interpretative strategies for lung function tests. PMID- 16264059 TI - Reversibility of pulmonary arterial hypertension in HIV/HHV8-associated Castleman's disease. AB - The present study describes a case of pulmonary arterial hypertension (PAH) associated with multicentric Castleman's disease in a patient infected with HIV type 1 and human herpes virus 8. Therapy included highly active antiretroviral therapy, warfarin, diuretics, continuous i.v. epoprostenol and 12-monthly pulses of cyclophosphamide. The patient's condition improved dramatically with complete reversibility of PAH, allowing weaning of continuous i.v. epoprostenol therapy. After 5 yrs, both Castleman's disease and PAH have not relapsed. This supports the hypothesis that control of inflammation and retroviral replication may be of interest in the context of PAH, complicating the course of an inflammatory condition associated with viral infection. In conclusion, further studies should help in characterising the best candidates for anti-inflammatory treatment in the setting of pulmonary arterial hypertension. PMID- 16264060 TI - A 3-year-old child with abdominal pain and fever. PMID- 16264061 TI - Optic neuritis with concurrent etanercept and isoniazid therapy. AB - OBJECTIVE: To report a case of optic neuritis associated with concurrent etanercept and isoniazid therapy. CASE SUMMARY: A 55-year-old man diagnosed as having rheumatoid arthritis had received treatment with nonsteroidal antiinflammatory drugs, sulfasalazine, oral methotrexate, leflunomide, and deflazacort. Four months prior to admission, he had a Disease Activity Score of 6.06; treatment with etanercept was considered. Three months prior to admission, because of evidence of latent tuberculosis, isoniazid 300 mg once daily and pyridoxine 50 mg once daily were prescribed. Treatment with subcutaneous etanercept 25 mg twice weekly was started 5 days after isoniazid was initiated. Two weeks prior to admission, the patient developed blurred vision in his left eye. Ten days later, his vision worsened and he was hospitalized. The visual acuity in both eyes was 0.7, and a campimetric examination was compatible with optic neuritis. Magnetic resonance imaging of the brain revealed lesions suggesting demyelinating lesions. The clinical course was consistent with bilateral optic neuritis. Etanercept was stopped, and isoniazid was replaced with rifampin 600 mg once daily. The patient was treated with intravenous methylprednisolone hemisuccinate 1 g/day for 5 days followed by oral prednisolone, resulting in a minor subjective improvement in left eye visual acuity. He then received oral prednisone for 3 weeks, slowly tapering to discontinuation. DISCUSSION: No physiologic factors could have predisposed this patient to develop optic neuritis. He was not diagnosed with a demyelinating disease or underlying systemic condition. The optic neuritis was unlikely to be an early manifestation of multiple sclerosis based on the clinical course and the negative results of the imaging tests. Furthermore, there was a close temporal correlation between the drug exposure and the onset of symptoms. After discontinuation of etanercept and isoniazid therapy, the patient's general condition improved. Use of the Naranjo probability scale indicated a possible relationship between optic neuritis and combined etanercept-isoniazid therapy. CONCLUSIONS: Patients initiated on etanercept and isoniazid should be closely monitored for the development of adverse neurologic signs and effects. If optic neuritis is determined, etanercept and isoniazid should be discontinued immediately. PMID- 16264062 TI - Effects of mometasone furoate given once daily in the evening on lung function and symptom control in persistent asthma. AB - BACKGROUND: The chronobiology of asthma suggests that, for once-daily dosing, an evening dose may be the most effective treatment paradigm. OBJECTIVE: To evaluate the efficacy and safety of mometasone furoate dry powder inhaler (MF-DPI) administered once daily in the evening or twice daily in patients with asthma previously maintained on twice-daily regimens of inhaled corticosteroids. METHODS: In this 12-week, multicenter, placebo-controlled trial, 268 subjects > or =12 years of age with inhaled corticosteroid-dependent asthma and baseline forced expiratory volume in 1 second (FEV(1)) between 50% and 85% of predicted were randomized to receive treatment with MF-DPI 400 mug once daily in the evening, MF-DPI 200 mug twice daily, or placebo. The primary efficacy variable was mean change in FEV(1) from baseline to endpoint. Other lung function measures, asthma symptoms, quality of life, and rescue medication use also were assessed. RESULTS: At endpoint, mean FEV(1) was significantly improved with both MF-DPI doses compared with placebo (p < 0.001). The 2 active treatment groups were statistically indistinguishable from each other. Secondary efficacy variables, including nocturnal awakenings, asthma worsenings, quality of life, and rescue medication use, were also significantly improved for both MF-DPI treatments compared with placebo. Both dosages were well tolerated; no clinically meaningful changes in laboratory values or vital signs were observed. CONCLUSIONS: MF-DPI 400 mug once daily in the evening was as effective as MF-DPI 200 mug twice daily in improving pulmonary function, asthma symptoms, and quality of life compared with placebo in subjects previously using twice-daily regimens of an inhaled corticosteroid. PMID- 16264063 TI - Role of botulinum toxin type A in the prophylactic treatment of migraine headaches. AB - OBJECTIVE: To evaluate the role of botulinum toxin type A in the prophylaxis of migraine headaches. DATA SOURCES: MEDLINE (1966-July 2005), Iowa Drug Information Service (1966-July 2005), and International Pharmaceutical Abstracts (1970-July 2005) searches were performed. Key search terms included botulinum and migraine. DATA SYNTHESIS: Patients who received botulinum toxin type A injections for treatment of hyperfunctional facial lines also experienced relief of migraine symptoms, leading to further investigation. Four clinical trials and 2 retrospective chart reviews evaluating the use of botulinum toxin type A injections for the prophylactic treatment of migraines are discussed. Limitations of these trials include problems with blinding, use of different injection sites, and inconsistent dosing. CONCLUSIONS: The results of 3 double-blind, placebo controlled trials, 2 retrospective chart reviews, and an open-label study indicate that botulinum toxin type A may be a safe and efficacious prophylactic treatment for migraine headaches. PMID- 16264064 TI - Cost of intravenous immunoglobulin therapy versus conventional immunosuppressive therapy in patients with mucous membrane pemphigoid: a preliminary study. AB - BACKGROUND: Intravenous immunoglobulin (IVIG) is an expensive biologic agent used to treat patients with mucous membrane pemphigoid (MMP) nonresponsive to conventional immunosuppressive therapy (CIST). The high cost of IVIG is of concern to healthcare providers and insurance companies. OBJECTIVE: To compare the cost of IVIG with that of CIST in treating a cohort of 15 patients with severe and extensive MMP. METHODS: Fifteen patients with biopsy-proven MMP nonresponsive to CIST were subsequently treated with IVIG and demonstrated a positive clinical response. This was a comparative, retrospective study; the mean total duration of the observation period was 8.4 years. A comparison of the cost of IVIG with that of CIST during the study period and the annual cost was performed. The cost of CIST was defined as the actual cost of the drug plus the cost of management of the multiple adverse effects, including hospitalizations, produced by CIST. In the same patient cohort, no significant adverse effects to IVIG were observed and no hospitalizations were required. Hence, the cost of IVIG therapy is simply the actual cost of the IVIG. RESULTS: In this cohort of patients, CIST had significant adverse effects, many of which were hazardous and required prolonged and frequent hospitalizations. The mean total cost using IVIG therapy was significantly less than that of CIST during the entire course of the disease (p < 0.001) and on an annual basis (p < 0.05). CONCLUSIONS: IVIG therapy is a safe, clinically beneficial, and less-expensive alternative treatment in patients with progressive MMP that is nonresponsive to CIST. PMID- 16264065 TI - The Ethical Conduct of Resuscitation Research: setting the stage for change. PMID- 16264066 TI - Executive summary: the 2005 AEM Consensus Conference on Ethical Conduct of Resuscitation Research. PMID- 16264067 TI - Keynote address: Toward a pragmatic model for community consultation in emergency research. AB - Under the Final Rule enacted in 1996 by the United States Food and Drug Administration and Office of Health and Human Services, community consultation and public notification are required when emergency research is to be conducted in the absence of prior informed consent by subjects. There is a dearth of published recommendations concerning the degree to which communities must be aware of the existence and parameters of a study for which informed consent might not be obtained prior to enrollment. It is argued that effective community consultation requires empirical measurement of the dynamics of community, and that ordinary notions of community may not capture the populations at greatest risk or those who might for other reasons figure most prominently in community consultation. A pragmatic approach to community consultation would establish benchmarks for such measures, and it is argued that such an approach is possible given the correct empirical measures. PMID- 16264068 TI - Current status of research on the federal guidelines for performing research using an exception from informed consent. AB - In order to set the stage for the later discussion of future directions, this Consensus Conference began by reviewing some of the existing research on conducting resuscitation studies using an exception from informed consent. That research is described in this article. The authors conclude that, based on the experiences reported by researchers and institutional review boards, resuscitation research may be ethically and legally conducted under the current regulations, but such research requires substantial effort, financial resources, and personnel. Federal regulations have set the bar appropriately high, and this may pose significant difficulties for small sponsors and individual investigators. However, further study of the experiences of subjects, researchers, and the regulatory process is warranted. PMID- 16264069 TI - Recruitment and communication process for participation in the 2005 AEM Consensus Conference on the Ethical Conduct of Resuscitation Research: methodology, challenges, lessons learned. AB - The 2005 Academic Emergency Medicine Consensus Conference, "Ethical Conduct of Resuscitation Research," was designed with the goal of developing consensus on important issues for human subjects and researchers surrounding the 1996 federal regulations jointly published by the Department of Health and Human Services and the Food and Drug Administration and known as the Final Rule. These regulations, which guide the conduct of research using the emergency exception from informed consent or waiver of informed consent, have been the subject of much debate in the resuscitation research community. Therefore, the editorial board of Academic Emergency Medicine chose this topic as the subject of their annual consensus conference. This report outlines the methods by which individuals and organizations were recruited to participate, how the conference was advertised, and the way in which participants and nonparticipants were encouraged to communicate before and after the conference. The limitations and potential biases of these methods and activities are also presented. PMID- 16264070 TI - Using the exception from informed consent regulations in research. AB - This article reflects the proceedings of a breakout session, "Using the Regulations in Research" at the 2005 Academic Emergency Medicine Consensus Conference, "Ethical Conduct of Resuscitation Research." There have been two organized studies, and a number of anecdotal reports, describing the decline in cardiac arrest resuscitation research in the United States since the implementation of the Final Rule. Paradis and colleagues found that the volume of human cardiac arrest research published in the United States was significantly less in a four-year period after the Final Rule was adopted as compared to the earlier period. Nichol and colleagues reported that both the absolute number of US-based randomized cardiac arrest trials and the proportion of US-based trials (vs. foreign trials, based on the mailing address of the first author) decreased by about 15% annually. Despite the concern about a negative impact, there are at least five published trials, one in progress and one in planning that have been or are being conducted under the regulations. Those completed include the Diaspirin Cross-Linked Hemoglobin, Public Access Defibrillation, Multicenter Vest CPR, Brain-CPR, and Pre-Hospital Treatment of Status Epilepticus trials. Reports of how investigators met the regulations and their experience in doing so are reviewed. A summary table of the federal regulations is provided. Participants discussed what additional information and research about using the regulations would be helpful for the promotion of quality resuscitation and emergency care research in the United States. Areas suggested for further investigation include: impact on the quality as well as quantity of such research; current level of understanding of the regulations by investigators, regulatory/IRB personnel and potential subjects (the general public); costs incurred: additional time required for preparation, approval and conducting community consultation and public disclosure; impact on research on non-life-threatening conditions; value and cost of a registry; use of a standard reporting template for issues regarding meeting the requirements in individual clinical trials; whether more specific guidance would be helpful or restrictive; what constitutes effective community consultation and public disclosure; and whether titration of community consultation and public disclosure based on the risk of the proposed intervention to subjects is feasible and acceptable. PMID- 16264072 TI - Researchers' understanding of the federal guidelines for waiver of and exception from informed consent. AB - The survival of patients who present to the emergency department with severe injury or illness is dismal. Resuscitation researchers are interested in advancing the science of resuscitation, and clinical studies must be conducted to determine the best treatment protocols. These studies must reflect good science and must balance individual patient autonomy and safety with scientific progress that benefits society as a whole. Researchers find the present federal guidelines on waiver of and exception from informed consent to be time consuming and expensive. They see variability in the requirements as interpreted by institutional review boards. There is confusion regarding the requirements for public notification and response to community consultation. They believe that the majority of the public, as well as health care professionals, want resuscitation research to progress, but a minority of people and governmental regulators are uncomfortable with waiver of and exception from informed consent for research studies. There is concern and some evidence that the federal guidelines have impeded the advancement of resuscitation science. Several strategies have been suggested to improve the situation. These include 1) better education of resuscitation researchers regarding the federal guidelines, 2) a toolbox for resuscitation researchers clarifying the guidelines, 3) advocacy for the advancement of resuscitation science as a public good, and 4) a national research advisory board that provides unbiased reviews of clinical studies and guidelines for local institutional review boards regarding risks, benefits, and communication strategies for waiver of and exception from consent proposals. PMID- 16264071 TI - Research conditions that qualify for emergency exception from informed consent. AB - Medical research involving critically ill and injured subjects unable to provide informed consent can only be conducted under federal regulations that attempt to balance the need to develop lifesaving treatments with protection of research subjects' rights. Regulators, researchers, and medical ethicists have all struggled to define the conditions under which an emergency exception from informed consent is appropriate. Although research has been successfully conducted under the current regulations, confusion remains regarding the meaning of the regulations, the applicable conditions, and the best ways to balance the needs of future patients and the rights of research subjects. In May 2005, at the Academic Emergency Medicine Consensus Conference "Ethical Conduct of Resuscitation Research," a breakout session was held on the research conditions that qualify for the emergency exception from informed consent process. Several recommendations emerged: 1) The definition of "life-threatening condition" should be broadly interpreted to include serious disability as well as death. 2) Existing therapies should be considered "unsatisfactory," even if partially effective, when serious risk of morbidity or mortality remains even with the best available treatment or when the adverse effects of the best available treatment are serious. 3) Research with the emergency exception should be performed only if sufficient evidence exists that the proposed intervention has a reasonable chance of benefit. 4) More evaluation is needed to determine the degree to which the current rules impede research. 5) Application of the current regulatory framework for abbreviated or waived consent in emergency research should be encouraged. 6) Further study should also address variability among institutional review boards, the goals of community involvement, and how best to engage and educate the public in research efforts using emergency exception from informed consent. PMID- 16264073 TI - Exception from informed consent: viewpoint of institutional review boards- balancing risks to subjects, community consultation, and future directions. AB - Differences in interpretation of the Final Rule for exception from informed consent (EFIC) requirements for emergency research result in inconsistencies in implementation and difficulties for some institutional review boards (IRBs) to approve such research. During a consensus workshop organized by the editorial board of Academic Emergency Medicine, participants discussed how IRBs balance the risks to human subjects in EFIC research, the conduct of community consultation and its role in IRB decision making, and future directions to improve and research EFIC effects. Areas of consensus and diversity of opinion were identified. During the workshop, the National Institutes of Health model of consensus building was used to develop statements pertaining to specific questions of the effects, directions, implementation, and ultimate goals for emergency research using EFIC. The program was composed of an overview of the history and issues related to EFIC or Final Rule research and presentations of viewpoints of experts in this area of research. A final consensus was developed regarding the major topics, including IRB perspective, effective community consultation (often considered the main difficulty in implementing EFIC research), and goals for future directions and research on the topic. Roundtable discussions and breakout sessions involving interested parties were used as a format. In regard to how IRBs balance risks, by consensus it was agreed the regulations stipulate that EFIC studies must involve treatment that is unproven or unsatisfactory. The committee agreed that resuscitation rates are currently unsatisfactory, and thus current treatments are unsatisfactory. Many treatments currently used as standard care have never been proven to be effective. IRBs and the public need education that resuscitation research is needed. The same can be said for other conditions to which this rule applies. Because IRB expertise differs across the country, a group of peer reviewers to act as consultants should be available to help IRBs determine if current treatment for a condition is unproven or unsatisfactory. In regard to community consultation, the experiences of others are important and helpful as guidance. The amount and formats of community consultation should correspond to the amount of risk involved in the study proposed. In regard to future directions, communities should be asked how they define "success" of community consultation and public disclosure. Research on community attitudes is critical. Ways to continue/add to research include the following: research including major National Institutes of Health/Centers for Disease Control and Prevention funding acquisition for evaluation of the clinical impact of EFIC research; education for research funding agencies about emergency research, including current outcomes (e.g., survival rates); participation of emergency medicine researchers in meetings of research ethicists/IRB members (Public Responsibility in Medicine and Research/Applied Research Ethics National Association); publication of experiences and of the effects of EFIC research; future update meetings such as this one at the Society for Academic Emergency Medicine meeting; and more membership on IRBs of emergency physicians. While IRBs must approve EFIC research based on their own local environment, additional guidelines from regulatory agencies may be helpful. In general, current treatments for EFIC conditions are unsatisfactory and many are unproven. A group of peer reviewers can act as consultants to IRBs that do not have this expertise. PMID- 16264074 TI - Does the emergency exception from informed consent process protect research subjects? AB - Although subject protection is the cornerstone of medical ethics, when considered in the context of research using emergency exception from informed consent, its success is debatable. The participants of a breakout session at the 2005 Academic Emergency Medicine Consensus Conference discussed the issues surrounding subject protection and advanced the following recommendations. 1) There are no outcome measures that define "protection"; therefore, it is not currently known whether or not subjects are protected under the current rules. 2) Care must be taken to protect not only the individual from harm during research but also to protect society from unregulated research in other countries and an inability to appropriately advance medical knowledge. 3) Some surrogate markers/methods of protection whose efficacies are debatable include data safety monitoring board activity, the community consultation and public notification (CC/PN) process, and institutional review board approval. 4) Minimal-risk studies should be held to different standards of protection than those that involve more significant risk to the subject. 5) A handful of studies have been published regarding community consultation and notification, and the majority are case studies. Those that are specifically designed to discover the most successful methods are hindered by a lack of formal outcomes measures and tend to have negative results. 6) Follow-up data from the CC/PN process should be disclosed to the Food and Drug Administration and incorporated into study designs. 7) Focus groups and/or random digit dialing have been suggested as promising methods for fulfilling the CC/PN requirements. 8) Studies need to be funded and performed that formally investigate the best means of CC/PN. 9) More funding for this research should be a priority in the emergency medicine and critical care communities. More data regarding terminated studies should be made available to the research community. 10) Quantifiable markers of success for CC/PN must be validated so that research may determine the most successful methods. 11) Data regarding subjects' and family members' experiences with exception from informed consent studies need to be obtained. PMID- 16264075 TI - Communicating with subjects: special challenges for resuscitation research. AB - In May 2005, Academic Emergency Medicine sponsored a one-day consensus conference held in association with the 2005 Society for Academic Emergency Medicine meeting in New York City. The conference, entitled "Ethical Conduct of Resuscitation Research," addressed a variety of issues regarding the successful conduct of research in acute care settings. A number of important breakout sessions were convened based on challenges specific to resuscitation research. "Communicating with Subjects" was one such session, dedicated toward perfecting the conduct of informed consent for research. The breakout session was attended by 15 insightful emergency medicine investigators, administrators, and ethicists. Issues of research information disclosure, subject comprehension, and the voluntariness of research participation were addressed. Consensus statements were developed and are discussed within this report. PMID- 16264076 TI - Communicating with communities about emergency research. AB - The challenge of effectively communicating with communities about research is particularly salient for investigators who are conducting emergency research with an exception from informed consent. The authors discuss the ethical basis for the community consultation requirement and describe the nature and extent of the consultative process required to achieve these ethical purposes. The findings of the consensus conference are summarized as follows. 1) The requirements for community consultation and public disclosure for exception from informed consent studies serve important ethical purposes and should be retained. 2) Community consultation allows investigators and institutional review boards to obtain input from the community regarding planned research. The process serves to facilitate understanding, promote trust, ensure justice, and protect research participants. 3) Community consultation is a process that requires active participation by community members; however, it does not require their approval, consent, or consensus. The practical challenges involved in conducting meaningful community consultation are also discussed: defining the community and its appropriate representatives, methods to actively engage the community, the lack of uniformity among institutional review boards in required community consultation activities, and the lack of measures to evaluate the adequacy and appropriateness of community consultation. The recommendations of the consensus conference regarding future research about community consultation are summarized. Priorities include identifying best practices; defining effectiveness and developing measures to evaluate community consultation; evaluating alternative models and potential infrastructures to facilitate, conduct, and/or oversee effective community consultation processes; and developing educational modules for community members to empower their active participation in discussions about emergency research in their communities. PMID- 16264077 TI - Resuscitation research involving vulnerable populations: are additional protections needed for emergency exception from informed consent? AB - The application of basic ethical principles to resuscitation research and other research in the emergency and critical care settings presents a particular challenge. The emergency exception from informed consent (EFIC) rule (21 CFR 50.24 and 45 CFR 46.101[i]) addresses a particular vulnerability: that which occurs when persons cannot consent due to acute loss of decisional capacity. The question arises as to whether populations defined as "vulnerable" are unique within studies to which EFIC applies. This report details the proceedings of a breakout session of the 2005 Academic Emergency Medicine Consensus Conference, "Ethical Conduct of Resuscitation Research," that dealt with this issue. Topics addressed were 1) the applicability of the term "vulnerable" in research using EFIC; 2) the relationship between vulnerability, exploitability, and capacity; 3) the significance of vulnerability in research design; 4) the adaptation of the informed consent process to the emergency setting; and 5) the role of the institutional review board. Ten consensus recommendations emerged from the discussion. Of particular importance was the endorsement of the idea that research using EFIC is as important in vulnerable populations as in the general population and that the systematic exclusion of vulnerable populations from resuscitation research is inappropriate. PMID- 16264078 TI - Confronting the Ethical Conduct of Resuscitation Research: a consensus opinion. AB - An objective of the 2005 Academic Emergency Medicine Consensus Conference, "Ethical Conduct of Resuscitation Research," was to identify if consensus exists regarding application of the Food and Drug Administration (FDA) Final Rule allowing an exception from informed consent in resuscitation research. At the start of the consensus conference, 49 attendees participated in a survey containing three sections: 1) demographic questions characterizing respondents, 2) questions regarding application of the FDA Final Rule, and 3) complexities associated with seeking informed consent in an emergency setting. Consensus analysis was used to determine if a formal consensus was reached, relying on a Bayesian posterior probability of 0.99 to consider survey responses a "consensus." Respondents demonstrated consensus regarding the need to further refine and standardize application of the FDA Final Rule in resuscitation research. However, participants agreed that current regulations provide adequate and appropriate protection to safeguard patients. Complexities associated with seeking informed consent in emergency departments were prevalent among most institutions represented at the conference. There was general agreement that current efforts to safeguard human subjects are effective, but participants agreed that refinements to and standardization of the FDA Final Rule would facilitate resuscitation research and enhance patient safety. PMID- 16264079 TI - Research without consent: community perspectives from the Community VOICES Study. AB - OBJECTIVE: To explore community attitudes toward the federal regulations that allow investigators to conduct emergency research without obtaining informed consent from participants. METHODS: Focus-group participants were recruited from residential sites in New York City that were enrolled in the Public Access Defibrillation (PAD) Trial. The PAD Trial, a National Institutes of Health funded, randomized trial in which laypersons were trained to treat cardiac arrest, was granted an exception from informed consent under these rules. Community residents and those who had been trained as lay responders in the PAD Trial were asked about the ethical issues raised by the conduct of research without consent (RWC), their definition of community, and appropriate methods of community consultation regarding RWC studies. Grounded theory content analyses were conducted on focus-group data. RESULTS: Seventeen (40%) men and 25 (60%) women from 15 buildings participated in six focus groups: four in English; two in Spanish. Definitions of "community" varied widely among and across groups; no strategy for community consultation was consistently endorsed by the participants. There were significant support and occasional adamant opposition to conducting RWC; participants often recounted specific personal experiences as the basis for both positive and negative opinions. Individuals with negative attitudes toward RWC often voiced strong support for specific RWC scenarios. CONCLUSIONS: There is no consensus regarding the definition of "community" or appropriate methods of consultation. Community members' attitudes toward RWC are often shaped by their personal experiences, and their general attitudes often differ from their reactions to specific RWC protocols. PMID- 16264080 TI - Impact of the Final Rule on the rate of clinical cardiac arrest research in the United States. AB - BACKGROUND: The federal government placed a moratorium on human emergency resuscitation research in 1993 due to concerns related to informed consent. In 1996, the Food and Drug Administration introduced the Final Rule in order to allow clinical resuscitation research to proceed in special cases without prospective informed consent. It is unclear, however, how this change has impacted the performance of this research. OBJECTIVES: To measure the potential impact of the Final Rule by determining the number of clinical cardiac arrest studies performed in the United States (US) before and after 1993, and to compare it with the number of studies performed in the European Union (EU) during the same periods. METHODS: A MEDLINE search was performed for cardiac arrest studies that were prospective, interventional, randomized, and performed in the US or EU from 1983 through 2003. RESULTS: Of the 3,115 cardiac arrest publications identified, 66 (2%) met inclusion criteria. Of the 40 studies conducted in the US, 24 (60%) were published prior to or during 1993, whereas of the 27 studies conducted in the EU, six (22%) were published prior to or during 1993 (odds ratio = 0.5; 95% confidence interval = 0.3 to 0.8). CONCLUSIONS: There has been a significant decrease in the number of published clinical cardiac arrest trials in the US since 1993. In contrast, there has been a significant increase in the number of published EU trials since 1993. The current informed consent requirements may have reduced the number of clinical cardiac arrest studies performed in the US. PMID- 16264081 TI - Responses to written notification during out-of-hospital care trials using waiver of informed consent. AB - BACKGROUND: Resuscitation research has been allowed to proceed using waiver of consent when compliance with guidelines is assured. In these circumstances, institutional review boards (IRBs) may request notification of enrolled patients. OBJECTIVES: To describe the notification process developed with IRB feedback and implemented in two out-of-hospital randomized controlled trials (RCTs) in a large urban setting. METHODS: This was a descriptive review of two RCTs recruiting a total of 620 patients: the Out-of-hospital Cardiac Arrest Rectilinear Biphasic to Monophasic Damped Sine Defibrillation Waveforms with Advanced Life Support Intervention Trial (ORBIT), which compared rectilinear biphasic and monophasic damped sine waveform for out-of-hospital cardiac arrest, and the Prehospital Pacing versus Conventional Treatment Effectiveness Trial (PrePACE), which compared dopamine and transcutaneous pacing for unstable bradycardia. RESULTS: The ORBIT study enrolled 538 patients. In 44 (8%) patients, contact information could not be obtained. Notification was attempted for 494 (92%) patients; 48 (10%) letters were returned to sender. Researchers were contacted by telephone regarding 37 (8%) subjects: 16 (43%) requested information, 14 (38%) responded positively, and one (3%) did not return follow-up calls. Overall, two (0.4%) were withdrawn from the study. The PrePACE study enrolled 82 subjects. Contact information was unavailable for one (1%). For the remaining 81 patients, four (5%) letters were returned to sender. Researchers were contacted regarding ten (13%) subjects: seven (70%) requested information, two (20%) responded positively, and one (10%) did not return follow-up calls. No patient was withdrawn from the study. CONCLUSIONS: Contact information may be difficult to obtain for up to 20% of out-of-hospital critical patients. In 8% to 13% of cases, patients or designates contacted researchers; most feedback was positive. Frequently, non-study-related information was requested, consuming significant resources. Using this method, fewer than 1% of patients were withdrawn from the studies. PMID- 16264082 TI - An ethical analysis of exception from informed consent regulations. AB - Federal regulations enacted in 1996 allow clinical research with an exception from informed consent in select emergency circumstances. Such research is only permitted when the therapy may directly benefit the research participant and it is not possible to conduct the research with informed consent. Among others, protective mechanisms include a process of community consultation and public disclosure and the establishment of a mandatory data safety monitoring board. Although the regulations do not preserve the right of self-determination of the research participant, the author argues that they do provide an ethical means to advance the science of resuscitation. PMID- 16264083 TI - Informed consent and ethical issues in military medical research. AB - Informed consent in military research shares many of the same fundamental principles and regulations that govern civilian biomedical research. In fact, much of modern research ethics is grounded in events that occurred in the context of war or government-sponsored research. Despite these similarities and common origins, research in the military has additional requirements designed to preserve service members' informed consent rights. The special nature of the superior-subordinate relationship in the military necessitates careful protections to avoid perceptions of coercion or undue influence on a military subject. Additionally, current legal and regulatory requirements for advanced informed consent significantly restrict the flexibility of the military to conduct research using waiver of consent. This has implications on the ability of the nation to develop effective medical treatments for the global war on terrorism. Nevertheless, work is under way to realign defense research policy with the norms of civilian biomedical practice. Future directions include the adoption of waivers for military emergency research, and the cautious introduction of human subject studies on the battlefield. This paper discusses historical background, regulatory differences, and concerns and challenges of some of these regulatory differences for research personnel that apply to informed consent and waiver of said informed consent for emergency research conducted by the U.S. military. PMID- 16264084 TI - Minimal-risk waiver of informed consent and exception from informed consent (Final Rule) studies at institutional review boards nationwide. AB - BACKGROUND: Federal regulations allow waiver of informed consent for minimal-risk research (waiver). More than minimal-risk emergency research is allowed with an exception from informed consent (EFIC) under specific guidelines called the Final Rule. Performance of research without consent is controversial; however, chances for public health improvements are significant. OBJECTIVES: 1) To describe the number and demographics of academic medical institutions that have reviewed and approved minimal-risk waiver and EFIC studies; 2) to describe associations between National Institutes of Health (NIH) funding percentile and whether an academic medical institution reviews EFIC studies; and 3) to describe associations between geographic region and review of EFIC studies, and the interaction of region and NIH funding percentile with review of EFIC studies. The hypothesis was that schools with a higher NIH funding percentile (more funding) reviewed more EFIC studies research, and that this effect did not vary based on region. METHODS: This was an e-mail and telephone survey of all 122 MD-degree granting medical schools in the continental United States. Descriptive data were collected on the number of schools reviewing and approving EFIC research. Sites were split into those that have reviewed EFIC studies and those that have not. A multivariable logistic regression examined the effects of 1) funding percentile and 2) region (Northeast, North Central, Southeast, and West) on the likelihood of EFIC study reviews at the sites. Odds ratios (ORs) and 95% confidence intervals (95% CIs) were determined. RESULTS: Ninety-eight of 118 eligible institutional review boards (IRBs) (83%) responded. Ninety-nine percent (97/98) approved waiver studies; 51% (50/98) had reviewed and approved EFIC studies. Academic medical institutions with higher funding percentile were significantly more likely to have reviewed/approved EFIC studies (interaction p = 0.01) in all regions except the Northeast, where higher funding percentile was associated with IRBs significantly less likely to have reviewed/approved EFIC studies (OR 0.25; 95% CI = 0.08 to 0.81). CONCLUSIONS: Almost all medical school IRBs review minimal-risk waiver studies. About half of IRBs at medical schools have reviewed and approved an EFIC study. The higher the NIH funding percentile (more NIH funding), the more likely a site has reviewed EFIC studies, except in the Northeast region. PMID- 16264086 TI - A single catalytic domain of the junction-resolving enzyme T7 endonuclease I is a non-specific nicking endonuclease. AB - A stable heterodimeric protein containing a single correctly folded catalytic domain (SCD) of T7 endonuclease I was produced by means of a trans-splicing intein system. As predicted by a model presented earlier, purified SCD protein acts a non-specific nicking endonuclease on normal linear DNA. The SCD retains some ability to recognize and cleave a deviated DNA double-helix near a nick or a strand-crossing site. Thus, we infer that the non-specific and nicked-site cleavage activities observed for the native T7 endonuclease I (as distinct from the resolution activity) are due to uncoordinated actions of the catalytic domains. The positively charged C-terminus of T7 Endo I is essential for the enzymatic activity of SCD, as it is for the native enzyme. We propose that the preference of the native enzyme for the resolution reaction is achieved by cooperativity in the binding of its two catalytic domains when presented with two of the arms across a four-way junction or cruciform structure. PMID- 16264088 TI - The role of nuclear factor kappa B in human labour. AB - Preterm birth remains the leading cause of perinatal mortality and morbidity, largely as a result of a poor understanding of the precise mechanisms controlling labour onset in humans. Inflammation has long been recognised as a key feature of both preterm and term labour, with an influx of inflammatory cells into the uterus and elevated levels of pro-inflammatory cytokines observed during parturition. Nuclear factor kappa B (NF-kappaB) is a transcription factor family classically associated with inflammation. Accumulating evidence points to a role for NF-kappaB in the physiology and pathophysiology of labour. NF-kappaB activity increases with labour onset and is central to multiple prolabour pathways. Premature or aberrant activation of NF-kappaB may thus contribute to preterm labour. The current understanding of NF-kappaB in the context of human labour is discussed here. PMID- 16264087 TI - Nearest-neighbor thermodynamics of deoxyinosine pairs in DNA duplexes. AB - Nearest-neighbor thermodynamic parameters of the 'universal pairing base' deoxyinosine were determined for the pairs I.C, I.A, I.T, I.G and I.I adjacent to G.C and A.T pairs. Ultraviolet absorbance melting curves were measured and non linear regression performed on 84 oligonucleotide duplexes with 9 or 12 bp lengths. These data were combined with data for 13 inosine containing duplexes from the literature. Multiple linear regression was used to solve for the 32 nearest-neighbor unknowns. The parameters predict the T(m) for all sequences within 1.2 degrees C on average. The general trend in decreasing stability is I.C > I.A > I.T approximately I. G > I.I. The stability trend for the base pair 5' of the I.X pair is G.C > C.G > A.T > T.A. The stability trend for the base pair 3' of I.X is the same. These trends indicate a complex interplay between H-bonding, nearest-neighbor stacking, and mismatch geometry. A survey of 14 tandem inosine pairs and 8 tandem self-complementary inosine pairs is also provided. These results may be used in the design of degenerate PCR primers and for degenerate microarray probes. PMID- 16264089 TI - Adipokines: implications for female fertility and obesity. AB - Obesity is associated with a diverse set of metabolic disorders, and has reproductive consequences that are complex and not well understood. The adipose tissue-produced leptin has dominated the literature with regards to female fertility complications, but it is pertinent to explore the likely role of other adipokines--adiponectin and resistin--as our understanding of their biological functions emerge. Leptin influences the developing embryo, the functioning of the ovary and the endometrium, interacts with the release and activity of gonadotrophins and the hormones that control their synthesis. In this review such biological actions and potential roles of the adipokines leptin, adiponectin and resistin are explored in relation to female fertility and the complexity of the obese metabolic state. PMID- 16264090 TI - Embryo culture does not affect the longevity of offspring in mice. AB - The oldest assisted reproductive technologies (ART)-conceived child is only 27 years old. Thus, the effects of ART on longevity are unknown, and it will be many years before this can be assessed in humans. We recently reported that culturing preimplantation mouse embryos under suboptimal conditions results in differences in how the offspring perform in behavioral assays that reflect anxiety (elevated zero maze) and spatial memory (Morris hidden water maze; Ecker et al. 2004). Here we monitored the mice generated in our previous study and found no difference in their longevity. PMID- 16264091 TI - Ontogeny and cellular localization of SRY transcripts in the human testes and its detection in spermatozoa. AB - The sex-determining region on the Y (SRY) gene is unequivocally designated as the testis-determining factor in mammals; however, its roles beyond sex determination, if any, have been hitherto unknown. To determine whether SRY has any roles beyond sex determination, herein the expression of SRY mRNA was investigated in the midtrimester human fetal, infantile and adult testes as well as in ejaculated spermatozoa. High levels of SRY transcripts were in situ localized to the Sertoli cells of the developing testis at 9 weeks of gestation, and the expression persisted at comparable levels throughout the midtrimester (until 22 weeks) and also in the testis of an infant at 3 months of age. The germ cells and other somatic cells in the testes of fetuses and the infant were negative for SRY expression. The mRNA for SRY was detected in the spermatogenic cells, particularly the spermatogonia and the round spermatids; the expression was negligible in the meiotic stages. A single transcript of approximately 1.2 kb was detected in the adult testes and isolated spermatogonial cells. In the adult testis, in situ hybridization (ISH) studies revealed a switch in the cellular localization of SRY transcripts. SRY transcripts were also demonstrable by RT-PCR of RNA from ejaculated human spermatozoa. ISH revealed the presence of SRY transcripts in the midpiece of 50% of ejaculated sperm. These results suggest that SRY may have extensive roles in male reproductive physiology, such as maturation of fetal testis, spermatogenesis, sperm maturation and early embryonic development. PMID- 16264092 TI - Localization of phosphatidylserine in boar sperm cell membranes during capacitation and acrosome reaction. AB - One of the essential properties of mammalian, including sperm, plasma membranes is a stable transversal lipid asymmetry with the aminophospholipids, phosphatidylserine (PS) and phosphatidylethanolamine (PE), typically in the inner, cytoplasmic leaflet. The maintenance of this nonrandom lipid distribution is important for the homeostasis of the cell. To clarify the relevance of lipid asymmetry to sperm function, we have studied the localization of PS in boar sperm cell membranes. By using labeled annexin V as a marker for PS and propidium iodide (PI) as a stain for nonviable cells in conjunction with different methods (flow cytometry, fluorescence and electron microscopy), we have assessed the surface exposure of PS in viable cells during sperm genesis, that is, before and during capacitation as well as after acrosome reaction. An approach was set up to address also the presence of PS in the outer acrosome membrane. The results show that PS is localized in the cytoplasmic leaflet of the plasma membrane as well as on the outer acrosome membrane. Our results further indicate the cytoplasmic localization of PS in the postacrosomal region. During capacitation and acrosome reaction of spermatozoa, PS does not become exposed on the outer surface of the viable cells. Only in a subpopulation of PI-positive sperm cells does PS became accessible upon capacitation. The stable cytoplasmic localization of PS in the plasma membrane, as well as in the outer acrosome membrane, is assumed to be essential for a proper genesis of sperm cells during capacitation and acrosome reaction. PMID- 16264093 TI - Different expression and activity of the alpha1 and alpha4 isoforms of the Na,K ATPase during rat male germ cell ontogeny. AB - Two catalytic isoforms of the Na,K-ATPase, alpha1 and alpha4, are present in testis. While alpha1 is ubiquitously expressed in tissues, alpha4 predominates in male germ cells. Each isoform has distinct enzymatic properties and appears to play specific roles. To gain insight into the relevance of the Na,K-ATPase alpha isoforms in male germ cell biology, we have studied the expression and activity of alpha1 and alpha4 during spermatogenesis and epididymal maturation. This was explored in rat testes at different ages, in isolated spermatogenic cells and in spermatozoa from the caput and caudal regions of the epididymis. Our results show that alpha1 and alpha4 undergo differential regulation during development. Whereas alpha1 exhibits only modest changes, alpha4 increases with gamete differentiation. The most drastic changes for alpha4 take place in spermatocytes at the mRNA level, and with the transition of round spermatids into spermatozoa for expression and activity of the protein. No further changes are detected during transit of spermatozoa through the epididymis. In addition, the cellular distribution of alpha4 is modified with development, being diffusely expressed at the plasma membrane and intracellular compartments of immature cells, finally to localize to the midregion of the spermatozoon flagellum. In contrast, the alpha1 isoform is evenly present along the plasma membrane of the developing and mature gametes. In conclusion, the Na,K-ATPase alpha1 and alpha4 isoforms are functional in diploid, meiotic and haploid male germ cells, alpha4 being significantly upregulated during spermatogenesis. These results support the importance of alpha4 in male gamete differentiation and function. PMID- 16264095 TI - Amino acid depletion and appearance during porcine preimplantation embryo development in vitro. AB - Preimplantation embryos can consume and produce amino acids in a manner dependent upon the stage of development that may be predictive of subsequent viability. In order to examine these relationships in the pig, patterns of net depletion and appearance of amino acids by in vitro produced porcine preimplantation embryos were examined. Cumulus oocyte complexes derived from slaughterhouse pre-pubertal pig ovaries were matured for 40 h in defined TCM-199 medium (containing PVA) before being fertilised (Day 0) with frozen-thawed semen in Tris-based medium. After 6 h, presumptive zygotes were denuded and cultured in groups of 20, in NCSU 23 medium modified to contain 0.1 mM glutamine plus a mixture of 19 amino acids (aa) at low concentrations (0.02-0.11 mM) (NCSU-23(aa)). Groups of 2-20 embryos were removed (dependent on stage) on Day 0 (1 cell), Day 1 (two- and four-cells), Day 4 (compact morulae) and Day 6 (blastocysts) and placed in 4 mul NCSU-23aa for 24 h. After incubation, the embryos were removed and the spent media was analysed by HPLC. The net rate of amino acid depletion or appearance varied according to amino acid (P < 0.001) and, apart from serine and histidine, stage of development (P < 0.014). Glycine, isoleucine, valine, phenylalanine, tryptophan, methionine, asparagine, lysine, glutamate and aspartate consistently appeared, whereas threonine, glutamine and arginine were consistently depleted. Five types of stage dependent trends could be observed: Type I: amino acids having high rates of net appearance on Day 0 that reached a nadir on Day 1 or 4 but subsequently increased by Day 6 (glycine, glutamate); Type II: those that exhibited lower rates of net appearance on Days 0 and 6 compared with the intermediate Days 1 and 4 (isoleucine, valine, phenylalanine, methionine, arginine); Type III: amino acids which showed a continuous fall in net appearance (asparagine, aspartate); Type IV: those that exhibited a steady fall in net depletion from Day 0 to Day 6 (glutamine, threonine); Type V: those following no discernable trend. Analysis of further embryo types indicated that presumptive polyspermic embryos on Day 0 had increased (P < 0.05) net rates of leucine, isoleucine, valine and glutamate appearance, and reduced (P < 0.05) net rates of threonine and glutamine depletion compared with normally inseminated oocytes. These data suggest that the net rates of depletion and uptake of amino acids by pig embryos vary between a) amino acids, b) the day of embryo development and, c) the type of embryos present at a given stage of development. The results also suggested that the net depletion and appearance rates of amino acids by early pig embryos might be more similar to those of the human than those of the mouse and cow. PMID- 16264094 TI - Estrogenic induction of spermatogenesis in the hypogonadal (hpg) mouse: role of androgens. AB - Testicular development is arrested in the hypogonadal (hpg) mouse due to a congenital deficiency of hypothalamic gonadotropin-releasing hormone synthesis. Previous studies have demonstrated that chronic treatment of these mice with estradiol induces testicular maturation and qualitatively normal spermatogenesis, but it is not known whether these are direct effects via estrogen receptors expressed in the testis, or indirect actions via the pituitary gland. The aim of the current studies was to determine whether the actions of estradiol require the presence of androgens. Sensitive assays revealed that chronic estradiol treatment produced time-dependent increases in pituitary FSH production but no increases in pituitary LH or testicular testosterone content could be detected. As a functional test of androgen dependence, hpg mice were treated for 70 days with estradiol plus Casodex (bicalutamide), an androgen receptor antagonist. Casodex treatment markedly attenuated both the estradiol-induced increase in testicular weight and the proliferation of the seminiferous epithelium, as revealed by morphometric analysis. However, it did not affect the estradiol-induced increase in pituitary FSH content, nor did it affect estradiol-induced increases in the weight of the seminal vesicles and epididymides. We conclude that increased FSH production is not sufficient to explain the increase in testicular development induced by estradiol in hpg mice; there is a requirement for functional androgen receptors for induction of testicular growth. PMID- 16264096 TI - Respiration rates of individual bovine in vitro-produced embryos measured with a novel, non-invasive and highly sensitive microsensor system. AB - Oxygen consumption is a useful parameter for evaluating embryo quality, since it provides a valuable indication of overall metabolic activity. Over the years, several approaches have been used to measure the respiration rates of individual embryos, but a convincing method has not yet been reported. In this study, we introduce and have validated a novel high resolution microsensor technology to determine the respiration rates of individual embryos at different developmental stages. We have employed this technology to investigate the correlation between respiration rate and embryo morphology, diameter and sex. Following morphological evaluation, individual respiration rates of day 3 (n = 18) and day 7 (n = 60) bovine in vitro-produced embryos were determined. Of the measured embryos, 64 were lysed for sex diagnosis by PCR. Average respiration rates of day 7 embryos (1.30 +/- 0.064 nl/h) were 3.4-fold higher than day 3 embryos (0.38 +/- 0.011 nl/h). On day 7, the average respiration rate of quality 1 blastocysts was significantly higher than the respiration rates of the lower qualities. For both day 3 and day 7 embryos, respiration rates were directly influenced by embryo diameter but did not differ between sexes. These results have demonstrated that the novel microsensor technology can be used to accurately and rapidly (8 min) measure the respiration rates of individual embryos at different developmental stages. Respiration rates were only in partial agreement with embryo morphology, suggesting a slight discrepancy between these two methods in assessing embryo quality. It is likely that a combined assessment of embryo respiration and morphology would improve embryo classification and subsequent selection. PMID- 16264097 TI - Morphological characterization of pre- and peri-implantation in vitro cultured, somatic cell nuclear transfer and in vivo derived ovine embryos. AB - The processes of cellular differentiation were studied in somatic cell nuclear transfer (SCNT), in vitro cultured (IVC) and in vivo developed (in vivo) ovine embryos on days 7, 9, 11, 13, 17 and 19. SCNT embryos were constructed from in vitro matured oocytes and granulosa cells, and IVC embryos were produced by in vitro culture of in vivo fertilized zygotes. Most SCNT and IVC embryos were transferred to recipients on day 6 while some remained in culture for day 7 processing. In vivo embryos were collected as zygotes, transferred to intermediate recipients and retransferred to final recipients on day 6. All embryos were processed for examination by light and transmission electron microscopy or immunohistochemical labelling for alpha-1-fetoprotein and vimentin. Overall, morphological development of in vivo embryos was superior to IVC and SCNT embryos. Day 7 and particularly day 9 IVC and SCNT embryos had impaired hypoblast development, some lacking identifiable inner cell masses. On day 11, only in vivo and IVC embryos had developed an embryonic disc, and gastrulation was evident in half of in vivo embryos and one IVC embryo. By day 13, all in vivo embryos had completed gastrulation whereas IVC and SCNT embryos remained retarded. On days 17 and 19, in vivo embryos had significantly more somites and a more developed allantois than IVC and SCNT embryos. We conclude that IVC and particularly SCNT procedures cause a retardation of embryo development and cell differentiation at days 7-19 of gestation. PMID- 16264098 TI - Peroxynitrites and impaired modulation of nitric oxide concentrations in embryos from diabetic rats during early organogenesis. AB - Maternal diabetes significantly increases the risk of congenital malformation, a syndrome known as diabetic embryopathy. Nitric oxide (NO), implicated in embryogenesis, has been found elevated in embryos from diabetic rats during organogenesis. The developmental signaling molecules endothelin-1 (ET-1) and 15 deoxy delta(12,14)prostaglandin J2 (15dPGJ2) downregulate embryonic NO levels. In the presence of NO and superoxide, formation of the potent oxidant peroxynitrite may occur. Therefore, we investigated peroxynitrite-induced damage, ET-1 and 15dPGJ2 concentrations, and the capability of ET-1, 15dPGJ2 and prostaglandin E2 (PGE2) to regulate NO production in embryos from severely diabetic rats (streptozotocin-induced before pregnancy). We found intense nitrotyrosine immunostaining (an index of peroxynitrite-induced damage) in neural folds, neural tube and developing heart of embryos from diabetic rats (P < 0.001 vs controls). We also found reduced ET-1 (P < 0.001) and 15dPGJ2 (P < 0.001) concentrations in embryos from diabetic rats when compared with controls. In addition, the inhibitory effect of ET-1, 15dPGJ2 and PGE2 on NO production found in control embryos was not observed in embryos from severely diabetic rats. In conclusion, both the demonstrated peroxynitrite-induced damage and the altered levels and function of multiple signaling molecules involved in the regulation of NO production provide supportive evidence of nitrosative stress in diabetic embryopathy. PMID- 16264099 TI - Spermatozoal competition in common carp (Cyprinus carpio): what is the primary determinant of competition success? AB - The percentage of sperm motility (92-100%), spermatozoan velocity (112-163 microm.s(-1)) and control hatching rates (83-96%) were evaluated for each of six gold and five green male common carp (Cyprinus carpio). In all 30 possible paired combinations of sperm-competition tests, hatching rates of 90-97% were achieved. The mean percentage of offspring sired was strongly influenced by the male used (P < 0.001, R2 = 0.91). The best male sired an average of 88% of the offspring in its competition tests, and the worst male sired only 5%. Spermatozoan-quality parameters could explain only part of the variation in male competitive ability. The male effects alone explained 91.4% of the observed variance, consisting of 17.1% explained by spermatozoan motility and 32.5% by control hatching rates in single fertilizations. Undetermined male effects explained 41.8%. The velocity of spermatozoa had no effect on the outcome of sperm competition. Neither was there any link between spermatozoan velocity and hatching rate in a control hatching test, whereas there was an effect of motility on hatching rate in this same test. PMID- 16264100 TI - Placentation in the African elephant, Loxodonta africana. IV. Growth and function of the fetal gonads. AB - The gonads, both ovaries and testes, of 44 elephant fetuses weighing 0.09-112 kg (6.1-21.3 months gestation) were examined grossly and histologically. As in equids, elephant fetal gonads undergo a phase of marked growth and enlargement during the second half of gestation, which is more pronounced in ovaries than testes due to growth and antrum formation of numerous follicles in the former. Stromal cells undergo hypertrophy and transformation to form zones of interstitial cells that are associated with the enlarged follicles in the ovaries and in which the primitive seminiferous tubules are embedded in the testes. The interstitial cells have the capacity to synthesize 5alpha-dihydroprogesterone and other 5alpha-reduced progestagens from cholesterol and pregnenelone and the hypothesis is raised that these fetal gonadal progestagens may supplement significantly the progestagens secreted by the multiple large corpora lutea of pregnancy in the elephant. PMID- 16264101 TI - Identification of genes regulated by interleukin-1beta in human endometrial stromal cells. AB - Interleukin-1beta (IL-1b) is an important immune regulatory factor that in human endometrium plays a role in both menstruation and implantation in the event of pregnancy. It promotes inflammatory-like processes and also stimulates tissue remodelling. We present a cDNA microarray study documenting the major effects of IL-1beta on gene expression in stromal cells from human endometrium. Endometrial stromal cells from five normal healthy women at the mid secretory phase were cultured with or without IL-1beta at 50 and 500 pg/ml for 48 h. cDNA microarrays were used to compare the levels of gene expression in total RNA isolated from cells stimulated with IL-1beta. These cDNA arrays were produced containing 15 164 sequence-verified clones, which included genes known to be important in angiogenesis, immune modulators, apoptosis, cell signalling, extra-cellular matrix (ECM) remodelling and cell cycle regulation. Genes which were regulated by IL-1beta were identified by analysis of the microarray data using the Significance Analysis of Microarrays software package. Upregulated (n = 23) and downregulated (n = 6) different genes were observed, which changed at least 3 fold, at a false discovery rate of less than 2% (P < 0.02). Our results have identified genes regulated by IL-1beta, which are involved in leukocyte recruitment, ECM remodelling and other cellular functions. Changes in three genes, IL-8, colony-stimulating factor 2 and aldoketo reductase family 1 member 1, which were upregulated by IL-1beta, were verified using real-time PCR. Novel functions regulated by IL-1beta in endometrium, including genes involved in free radical protection, and fatty acid metabolism were also identified. These results also provide new insights into the role of IL-1beta in disorders of the endometrium, especially in implantation-related infertility and endometriosis, in which this cytokine plays a major role. PMID- 16264102 TI - Effects of long-term maternal exposure to low doses of PCB126 and PCB153 on the reproductive system and related hormones of young male goats. AB - In this study, female goats were orally exposed to PCB126 or PCB153, at 49 ng/kg body weight per day and 98 microg/kg body weight per day respectively, from gestational day 60 until delivery at approximately day 150. Exposure of the offspring continued via lactation until postnatal day 40. Reproductive toxicity in the male offspring was studied by the evaluation of conventional reproductive endpoints as well as flow cytometric analyses of spermatogenesis and sperm chromatin structure. PCB153-treated animals showed a significant smaller testis diameter in comparison to the control group. Neither of the treated groups showed differences for plasma FSH in comparison to controls. PCB153-treated animals differed significantly from the control group with respect to plasma LH and testosterone levels, whereas PCB126-treated animals only differed from the controls in plasma testosterone concentrations. Neither the PCB126 nor the PCB153 group differed from the controls with respect to the conventional sperm parameters or testis histology. A significant lower ratio of interstitium area to seminiferous tubules area and proportion of diploid testis cells were observed for the PCB153 group. Sperm from PCB153-treated animals showed a significantly higher percentage of sperm with damaged DNA. From the results of the present study it was concluded that PCB153 was able to induce alterations in reproductive endpoints related to the hypothalamic-pituitary-axis as well as to the testis. The effects observed in male kids after a long-term maternal exposure to PCB153 support the concept that exposure to endocrine-disrupting compounds during foetal development may lead to adverse reproductive effects in adult life. PMID- 16264103 TI - A rat model for functional characterization of pregnancy-induced denervation and postpartum reinnervation in the myometrium and cervix: a superfusion study. AB - The pregnancy-induced rapid degeneration of the adrenergic nerves innervating the uterus is a well-known but poorly understood phenomenon. Since most of the published investigations were carried out by histological assay, our aim was to describe the loss of the adrenergic function during pregnancy and the re innervational procedure in the postpartum period. Myometrial and cervical samples from rats were loaded with [3H]noradrenaline and then transferred into a chamber for superfusion. After a wash-out period, fractions were collected. The fifth and fifteenth fraction tissues were stimulated with an electric field. The [3H]noradrenaline contents of the fractions were determined, together with the amount remaining in the tissue. The myometrial [3H]noradrenaline release was substantially decreased in early pregnancy, and absent in the late stage. Differences in release profile were detected between the implantation sites and the interimplantation areas. As a refinement of the results of previous histochemical studies, the noradrenergic functions of the cervix were found to be deeply affected in the early postpartum period. The pregnancy-induced denervational procedure can be followed by means of a superfusional technique after [3H]noradrenaline loading. As our technique is considered to be similar in sensitivity to histological methods, superfusion can be regarded as a model for functional investigations of pregnancy-induced denervation. PMID- 16264105 TI - Bisphosphonates in metastatic bone disease: renal safety matters. PMID- 16264104 TI - Apoptosis of bovine ovarian surface epithelial cells by Fas antigen/Fas ligand signaling. AB - Ovarian surface epithelial cells (OSEs), a single layer of cells that cover the surface of the ovary, undergo turnover at the site of follicular rupture at ovulation. Greater than 90% of ovarian cancers arise from the OSEs. The objective of this study was to determine whether OSEs have the capacity to regulate their own demise through expression of Fas antigen (Fas) and Fas ligand (FasL) and activation of Fas-mediated apoptosis. In initial experiments, primary cultures of bovine OSEs responded to treatment with recombinant FasL by undergoing apoptosis. The percentage of cell death was not affected by the presence or absence of serum in the media or by co-treatment with interferon-gamma, a treatment shown to potentiate Fas-mediated apoptosis in a number of cell types. Subsequent experiments tested the ability of stress-inducing drugs, anisomycin and daunorubicin, to promote apoptosis by stimulating an endogenous Fas-FasL pathway in OSEs. Treatment with FasL, anisomycin or daunorubicin induced cell death and this was suppressed by co-treatment with a peptide inhibitor of caspases, ZVAD. Treatment with anisomycin or daunorubicin in the presence of ZVAD increased expression of FasL mRNA and protein but did not alter expression of Fas mRNA or protein. Treatment of OSEs with a recombinant protein that blocks interaction of FasL with Fas (Fas:Fc) reduced apoptosis in response to anisomycin and daunorubicin, indicating that drug-induced apoptosis was mediated at least partially through endogenous Fas-FasL interactions. In summary, OSEs undergo apoptosis in response to stress-inducing drugs through activation of an endogenous Fas pathway. PMID- 16264106 TI - Preclinical perspectives on bisphosphonate renal safety. AB - Renal insufficiency is not rare in cancer patients who may receive nephrotoxic medications as antineo-plastic agents or for comorbid conditions. Thus, the choice of a particular bisphosphonate for patients with metastatic bone disease should be based not only on efficacy but also on the risk for renal deterioration. Some i.v. bisphosphonates have been associated with occasional renal toxicity in the clinical setting. Preclinical studies have also shown that there may be considerable differences among bisphosphonate renal safety profiles. Comparative studies show variations in the risk for histopathologic damage and the ability to cause cumulative toxicity during intermittent dosing. Reasons for the differences among bisphosphonates are not fully understood; however, research shows that they may be influenced by pharmacokinetic properties such as renal tissue half-life or protein binding and intracellular potency. Further preclinical analyses are needed to confirm and evaluate differences among bisphosphonates. PMID- 16264107 TI - Efficacy of ibandronate in metastatic bone disease: review of clinical data. AB - Metastatic bone disease affects many cancer patients and has a significant disease burden because of complications such as pathologic fractures and severe pain, which affect patient mobility and quality of life. Bisphosphonates are the current standard of care for treating metastatic bone disease. Available agents have shown varying degrees of efficacy in clinical trials, and treatment potential can be limited by efficacy, tolerance, or toxicity issues. Ibandronate (Bondronat); F. Hoffman-La Roche Ltd., Basel, Switzerland, http://www.roche.com) is a highly potent, single-nitrogen bisphosphonate that is available in i.v. and oral formulations. In phase III trials in breast cancer patients, both formulations reduced the incidence of skeletal complications associated with metastatic bone disease and had significant and sustained effects on bone pain and patient quality of life. Open-label studies of loading-dose ibandronate administered over consecutive days suggest it also may be useful for relieving severe or opioid-resistant metastatic bone pain. New trials have been designed or are in progress that may extend the clinical indications of ibandronate for patients with metastatic bone disease. PMID- 16264108 TI - Renal safety of ibandronate. AB - Despite their efficacy in treating complications associated with metastatic bone disease, there are concerns about the potential nephrotoxicity of certain i.v. bisphosphonates for the long-term management of cancer patients. Clinical data suggest, however, that i.v. ibandronate (Bondronat); F. Hoffman-La Roche Ltd., Basel, Switzerland, http://www.roche.com), unlike other bisphosphonates, has a renal safety profile comparable with that of placebo. In a 2-year, phase III study of patients with breast cancer metastatic to bone, the incidence of adverse renal events in patients treated with 6 mg i.v. ibandronate was low and comparable with that of placebo (4% versus 4.5% with placebo). Two-year assessments of time to serum creatinine increase also demonstrated renal safety comparable with that of placebo (patients with creatinine increase: 6% versus 12% with placebo). Long-term (4-year) renal safety of ibandronate was demonstrated in a 2-year extension of the trial. Phase II, open-label studies show that intensive ibandronate dosing does not compromise renal safety in patients with metastatic bone pain from a variety of tumor types. In addition, i.v. ibandronate is well tolerated, with no evidence of renal toxicity in multiple myeloma and urologic cancer patients with existing renal impairment. The potential nephrotoxicity of some bisphosphonates has prompted additional renal safety precautions in product labeling for these agents. The precautions are not, however, contained in the label for ibandronate, which may thus simplify patient management. PMID- 16264109 TI - Bisphosphonate treatment recommendations for oncologists. AB - Renal safety is an important consideration for oncologists who are treating patients with bisphosphonates. In recent years, there has been increasing awareness about the development of bisphosphonate-induced nephrotoxicity. This has emerged mainly from increased clinical experience with zoledronic acid (Zometa); Novartis Pharmaceuticals Corporation, East Hanover, NJ, http://www.pharma.us.novartis.com). For this reason, the U.S. and European product labels for i.v. zoledronic acid were recently updated to include additional renal safety cautions, including dose adjustment in patients with mild to-moderate renal impairment. However, renal toxicity is not a class effect. The product label for ibandronate (Bondronat), F. Hoffmann-La Roche Ltd., Basel, Switzerland, http://www.roche.com) has remained unchanged since the launch of the drug in the European Union in 2003. Ibandronate does not require mandatory monitoring of kidney function prior to each infusion. In addition, ibandronate can be used in patients with varying degrees of renal impairment. It also can be used without restrictions for nephrotoxic medications, and dose adjustment is only required in patients with severe renal impairment. Clinical implications of the renal safety of ibandronate include reducing the physician and nursing time needed for managing the adverse renal events associated with bisphosphonate therapy and dosing based on renal function. There also are no added renal safety risks and fewer inconvenient hospital visits with ibandronate therapy. In addition to i.v. ibandronate, an oral formulation of the drug is available. Oral ibandronate therapy is especially desirable because the medication is convenient (with a small, once-daily tablet that can be taken at home), reducing the health care costs associated with infusions. Clinical studies also indicate that 50 mg oral ibandronate has an efficacy similar to that of i.v. bisphosphonates and is associated with a low incidence of adverse gastrointestinal events. PMID- 16264110 TI - Physeal sparing reconstruction of the anterior cruciate ligament in skeletally immature prepubescent children and adolescents. AB - BACKGROUND: The management of anterior cruciate ligament injuries in skeletally immature patients is controversial. Conventional adult reconstruction techniques risk potential iatrogenic growth disturbance due to physeal damage. The purpose of this study was to evaluate the results of a physeal sparing, combined intra articular and extra-articular reconstruction technique in prepubescent skeletally immature children. METHODS: Between 1980 and 2002, forty-four skeletally immature prepubescent children and adolescents who were in Tanner stage 1 or 2 (with a mean chronological age of 10.3 years) underwent physeal sparing, combined intraarticular and extra-articular reconstruction of the anterior cruciate ligament with use of an autogenous iliotibial band graft. Twenty-seven patients had additional meniscal surgery. Functional outcome, graft survival, radiographic outcome, and growth disturbance were evaluated at a mean of 5.3 years after surgery. RESULTS: Two patients underwent a revision reconstruction for graft failure at 4.7 and 8.3 years postoperatively. In the remaining forty-two patients, the mean International Knee Documentation subjective knee score (and standard deviation) was 96.7 +/- 6.0 points, and the mean Lysholm knee score was 95.7 +/- 6.7 points. The results of the Lachman examination for anterior cruciate ligament integrity was normal for twenty-three patients, nearly normal for eighteen patients, and abnormal for one patient. The results of the pivot-shift examination were normal for thirty-one patients and nearly normal for eleven patients. Four of the twenty-three patients who underwent concurrent meniscal repair had a repeat arthroscopic meniscal repair or partial meniscectomy. The mean growth in total height from the time of surgery to the final follow-up evaluation was 21.5 cm. No patient had an angular deformity measured radiographically or a discrepancy in the length of the lower extremities measured clinically. CONCLUSIONS: Physeal sparing, combined intra-articular and extra articular reconstruction of the anterior cruciate ligament with use of an autogenous iliotibial band graft in skeletally immature prepubescent children and adolescents provides excellent functional outcome with a low revision rate and a minimal risk of growth disturbance. PMID- 16264111 TI - Treatment for osteonecrosis of the femoral head: comparison of extracorporeal shock waves with core decompression and bone-grafting. AB - BACKGROUND: There is continuing controversy regarding the optimal treatment for patients with symptomatic early-stage osteonecrosis of the femoral head. We compared the results of noninvasive treatment with extracorporeal shock waves with those of core decompression and bone-grafting in similar groups of patients. METHODS: Patients with stage-I, II, or III osteonecrosis were randomly assigned to be treated either with shock waves or with core decompression and nonvascularized fibular grafting. The shock-wave group consisted of twenty-three patients (twenty-nine hips), and the surgical group consisted of twenty-five patients (twenty-eight hips). The patients in the two groups had similar demographic characteristics, duration and stage of disease, and duration of follow-up. The patients in the shock-wave group received a single treatment with 6000 impulses of shock waves at 28 kV to the affected hip. The evaluation parameters included clinical assessment of pain with a visual analog pain scale, Harris hip scores, and an assessment of activities of daily living and work capacity. Radiographic assessment was performed with serial plain radiographs and magnetic resonance imaging. RESULTS: Before treatment, the two groups had similar pain and Harris hip scores. At an average of twenty-five months after treatment, the pain and Harris hip scores in the shock-wave group were significantly improved compared with the pretreatment scores (p < 0.001). In this group, 79% of the hips were improved, 10% were unchanged, and 10% were worse. Of the hips treated with a nonvascularized fibular graft, 29% were improved, 36% were unchanged, and 36% were worse. In the shock-wave group, imaging studies showed regression of five of the thirteen lesions that had been designated as stage I or II before treatment and no regression of a stage-III lesion. Two stage-II and two stage-III lesions progressed. In the surgical group, four lesions regressed and fifteen (of the nineteen graded as stage I or II) progressed. The remaining nine lesions were unchanged. CONCLUSIONS: Extracorporeal shock-wave treatment appeared to be more effective than core decompression and nonvascularized fibular grafting in patients with early-stage osteonecrosis of the femoral head. Long-term results are needed to determine whether the effect of this novel method of treatment for osteonecrosis of the femoral head endures. PMID- 16264112 TI - Helical computed tomography alone compared with plain radiographs with adjunct computed tomography to evaluate the cervical spine after high-energy trauma. AB - BACKGROUND: Current literature supports the use of the three-view plain radiograph series supplemented, when necessary, with helical computed tomography to evaluate the cervical spine in patients who have sustained trauma injury. The purpose of this study was to determine if helical computed tomography alone can be used to evaluate the cervical spine for acute osseous injury following high energy trauma, thus eliminating the need to make radiographs. METHODS: Patients were prospectively evaluated with helical computed tomography scanning of the cervical spine and standard three-view plain radiography. At a later date, the plain radiographs and computed tomography scans were independently reviewed by two radiologists who were blinded to both the initial interpretation and the interpretation of the corresponding study. The radiologists documented whether the plain radiographs were adequate and whether they showed an acute process. The findings in the study were compared with the initial findings and, when necessary, with the discharge summaries to determine if an injury had been identified. The accuracy of the plain radiographs, of the plain radiographs that had been deemed adequate, and of helical computed tomography used alone was ascertained. RESULTS: Plain radiographs and helical computed tomography scans were made for 407 patients, and traumatic injuries were identified in fifty-eight of them. Plain radiographs alone were adequate for 194 (48%) of the 407 patients. Plain radiographs had a sensitivity of 45%, a specificity of 97%, a positive predictive value of 74%, and a negative predictive value of 91%. Adequate plain radiographs had a sensitivity of 52%, a specificity of 98%, a positive predictive value of 81%, and a negative predictive value of 93%. Helical computed tomography had a sensitivity and specificity of 98%, a positive predictive value of 89%, and a negative predictive value of >99%. The sensitivity, positive predictive value, and negative predictive value of adequate plain radiographs differed significantly from those of helical computed tomography alone (p < 0.001). Twelve (48%) of twenty-five adequate plain radiographs of patients in whom an injury had been identified on computed tomography missed that injury. Helical computed tomography alone missed one (2%) of the fifty-eight injuries. CONCLUSIONS: Although helical computed tomography has a limited ability to detect pure ligamentous injury, it can be safely used without plain radiographs to evaluate the cervical spine for osseous abnormalities such as fractures and dislocations after high-energy trauma. PMID- 16264113 TI - The medical-legal aspects of informed consent in orthopaedic surgery. AB - BACKGROUND: Orthopaedic surgeons routinely obtain informed consent prior to surgery. Legally adequate informed consent requires a thorough discussion of treatment options and risks and proper documentation; however, there is little data to guide orthopaedic surgeons regarding effective methods of obtaining informed consent. METHODS: We performed a closed claims analysis on malpractice claims involving an allegation of inadequate informed consent brought during a twenty-four-year period with two malpractice insurers. Relevant malpractice claims were reviewed, and data were abstracted. We then performed statistical analyses to identify factors that positively correlated with a successful defense. RESULTS: We identified twenty-eight lawsuits that included a claim of inadequate informed consent. All of the cases involved elective orthopaedic surgical procedures; there were no emergent cases. Three cases involved a disputed surgical site; all three cases involved foot and ankle surgery and resulted in an indemnity payment. Documentation of appropriate informed consent in the office notes of the surgeon was associated with a decreased indemnity risk (p < 0.005). Obtaining the informed consent on the hospital ward or in the preoperative holding area was associated with an increased indemnity risk (p < 0.004). When informed consent was obtained in the office by the operating surgeon, the risk of malpractice payment was significantly decreased (p < 0.004). CONCLUSIONS: Surgeons may be able to decrease the risk of a malpractice claim by obtaining informed consent in their offices, rather than in the preoperative holding area, and by documenting the informed consent discussion within their dictated office or operative notes. PMID- 16264114 TI - Enhancement of tendon-bone healing of anterior cruciate ligament grafts by blockage of matrix metalloproteinases. AB - BACKGROUND: The use of soft-tissue grafts for anterior cruciate ligament reconstruction delays the healing process. This delay may be due to biochemical and/or biomechanical insults. We hypothesized that the blocking effect of alpha2 macroglobulin on synovial matrix metalloproteinase activity may enhance the healing of tendon graft in a bone tunnel. METHODS: The study was performed on twenty-eight healthy, skeletally mature New Zealand White rabbits. Each rabbit underwent bilateral anterior cruciate ligament reconstruction with use of the ipsilateral semitendinosus tendon. Alpha-2-macroglobulin (alpha2-macroglobulin) was injected into the knee joint in one limb, and the contralateral limb served as a control. The rabbits were killed two weeks (fourteen rabbits) or five weeks (fourteen rabbits) after the operative procedures. The presence of matrix metalloproteinases in synovial fluid, and the blocking effect of alpha2 macroglobulin on them, were determined with enzymatic assays. Healing between the tendon and the bone tunnel was assessed morphologically by determining the presence of fibrovascular tissue and collagen fibers. Healing also was assessed quantitatively by measuring the ultimate load to failure of the reconstructed complex. RESULTS: There was an increase in matrix metalloproteinases in the control group; in contrast, there was a decrease in the study group (p < 0.05). In the control specimens, the fibrovascular tissue at the bone-tendon interface had developed into dense connective tissue with poor vascularization. In the treated specimens, the bone tunnel had more areas of denser connective-tissue ingrowth. The interface tissue was more mature and contained numerous perpendicular collagen bundles (Sharpey fibers). The ultimate load to failure was significantly greater in the alpha2-macroglobulin-treated specimens than in the untreated controls at both two and five weeks. CONCLUSIONS: The present study demonstrated that alpha2-macroglobulin blockade of matrix metalloproteinases can enhance bone-tendon healing. This effect of alpha2-macroglobulin could occur through its effect solely on collagenase or on a subset of matrix metalloproteinases that are present at the healing interface. PMID- 16264115 TI - The effect of femoral notching during total knee arthroplasty on the prevalence of postoperative femoral fractures and on clinical outcome. AB - BACKGROUND: The treatment of a supracondylar femoral fracture following total knee arthroplasty is complicated by the presence of the prosthetic components. Anterior femoral notching during arthroplasty has been implicated as a contributing risk factor for femoral fracture. We retrospectively reviewed the effect of anterior femoral notching on the subsequent occurrence of a periprosthetic supracondylar fracture of the distal aspect of the femur and the outcomes of primary total knee arthroplasty in such patients. METHODS: The prevalence and depth of femoral notching were determined on a review of the lateral radiographs by observers blinded to the clinical results of 1089 consecutive total knee replacements performed in 1997 and 1998. Linear and logistic regression modeling was used to analyze the relationship between femoral notching and the prevalence of supracondylar femoral fracture, postoperative range of motion, the Knee Society score, and the Knee Society functional and pain scores. RESULTS: Femoral notching was performed in 325 (29.8%) of the 1089 knees in our series. During an average follow-up period of 5.1 years, only two supracondylar femoral fractures occurred, both in femora treated without notching. Femoral notching was not associated with an increased rate of fracture (p = 1.000) or with significant differences in the measures of outcome (range of motion [p = 0.117], knee score [p = 0.967], functional score [p = 0.861], need for a lateral release [p = 0.234], or postoperative pain [p = 0.948]). CONCLUSIONS: This study demonstrated no difference in knees managed with or without notching of the anterior distal aspect of the femur with respect to the occurrence of a supracondylar fracture, range of motion, Knee Society score, Knee Society function, or pain. PMID- 16264116 TI - Accuracy in the measurement of compartment pressures: a comparison of three commonly used devices. AB - BACKGROUND: In situations in which accurate physical diagnosis is inconclusive, an objective method for measuring compartment pressure can aid in the diagnosis of compartment syndrome. Previous studies have compared measurement devices with each other but not with an accurately determined gold standard. The purpose of the present study was to devise a reproducible in vitro model of compartment pressure and to compare commonly used measurement devices in order to determine their accuracy. METHODS: With a graduated cylinder being used to generate a known pressure, freshly harvested ovine muscle was placed into a chamber for testing. The cylinder was incrementally filled with saline solution (in fifty-five steps), and measurements of tissue pressure were obtained with use of the Stryker Intracompartmental Pressure Monitor System, an arterial line manometer, and the Whitesides apparatus. Each device was tested with a straight needle, a side-port needle, and a slit catheter, for a total of nine setups in all. Five trials were done with each setup. Control pressures were calculated on the basis of the height of the saline solution column (test range, 0.13 to 10.80 kPa). Multiple regression analysis was used to compare measured tissue pressures with calculated control pressures. RESULTS: Most methods demonstrated excellent correlation (R2> 0.95) between calculated and measured pressures. The arterial line manometer with the slit catheter showed the best correlation (R2= 0.9978), and the Whitesides apparatus with the side-port needle showed the worst (R2= 0.9115). Furthermore, the Stryker system with the side-port needle demonstrated the least constant bias (+0.06 kPa). Straight needles tended to overestimate pressure. Two of the three needle configurations involving the Whitesides apparatus overestimated pressure. The data for the Whitesides methods had the highest standard errors, showing clinically unacceptable scatter. CONCLUSION: Side-port needles and slit catheters are more accurate than straight needles are. The arterial line manometer is the most accurate device. The Stryker device is also very accurate. The Whitesides manometer apparatus lacks the precision needed for clinical use. PMID- 16264117 TI - Clinical outcomes of unstable pelvic fractures in skeletally immature patients. AB - BACKGROUND: The orthopaedic literature contains few studies evaluating the long term outcomes of unstable pelvic fractures in skeletally immature patients. The purpose of this study was to determine the factors that may influence the clinical and functional outcomes of such fractures. METHODS: A retrospective review of all patients with open triradiate cartilages and an unstable pelvic (Tile type-B or C) fracture treated, from 1986 to 2000, at one of two level-I trauma centers was performed. Patients were evaluated with a review of their medical records, the Modified Injury Severity Score (MISS), standardized physical examination, standardized radiographic evaluation, and the Short Musculoskeletal Function Assessment Questionnaire (SMFA). The outcomes were then used to assess the difference between patients who had been treated operatively and those who had been treated nonoperatively. RESULTS: Of 230 pelvic fractures treated during the study period, twenty-three in twenty-three patients were unstable. Of the twenty-three patients, twenty, with a mean age of 9.5 years at the time of injury, were evaluated. The mean duration of follow-up was 6.5 years. There were four type-B and sixteen type-C fractures according to the Tile classification system. The four patients with a type-B fracture had a mean of 1.4 cm of pelvic asymmetry at the time of union and the last follow-up, whereas the sixteen patients with a type-C fracture had a mean of 1.5 cm of pelvic asymmetry at those times. Pelvic asymmetry did not remodel even in younger patients. Eighteen patients were treated operatively with external fixation, internal fixation, or a combination of both, and pelvic asymmetry of < or =1 cm was achieved in ten of them. Patients who had < or =1 cm of pelvic asymmetry had no lumbar or sacroiliac pain, no or mild sacroiliac tenderness, no Trendelenburg sign, no lumbar scoliosis, and lower (better) bother and dysfunction scores on the SMFA compared with patients with more pelvic asymmetry. All patients with > or =1.1 cm of pelvic asymmetry had three or more of the following: nonstructural scoliosis, lumbar pain, a Trendelenburg sign, or sacroiliac joint tenderness and pain. Patients with fewer associated injuries and pelvic asymmetry of < or =1 cm had better clinical results. CONCLUSIONS: Unstable pelvic fractures in children can result in long-term morbidity and functional problems. Fractures associated with > or =1.1 cm of pelvic asymmetry following closed reduction should be treated with open reduction and internal or external fixation in order to improve alignment and the long-term functional outcome. PMID- 16264118 TI - Early complications of primary total hip replacement performed with a two incision minimally invasive technique. AB - BACKGROUND: Total hip replacement performed through a small incision theoretically results in less trauma to the underlying structures, reduced blood loss, less pain, and a shorter hospital stay, but it may result in increased complications, particularly early in a surgeon's experience with a new technique. In the present study, we reviewed the early results of two techniques involving the use of smaller incisions; specifically, we evaluated one series of primary total hip replacements that had been performed through two small incisions and another series of total hip replacements that had been performed through a single small incision. METHODS: Eighty-nine consecutive primary total hip replacements were performed with use of the two-incision technique as described by Mears and Berger; all procedures were performed without cement and with use of fluoroscopic guidance. Outcomes data were reviewed at a minimum of six months following the procedure. The results of these procedures were retrospectively compared with those of a historical control series of ninety-six total hip replacements that had been performed by the same surgeon with use of a single mini-incision technique. No special attempt was made to discharge any patient early from the hospital. In preparation for the use of the two-incision technique, the surgeon attended a two-day seminar that included cadaveric training and mentoring by surgeons who had experience with this technique. RESULTS: In the two-incision group, nine patients (nine hips; 10%) required repeat surgery because of a femoral fracture that had been identified postoperatively (two hips), dislocation (one hip), a wound complication (two hips), or subsidence and loosening of the femoral implant (four hips). Twenty-two patients (twenty-two hips; 25%) sustained an injury of the lateral femoral cutaneous nerve, and one patient (one hip) had a neuropraxia of the femoral nerve. In the comparative series of ninety-six total hip arthroplasties that had been performed with use of a single mini-incision and a direct lateral exposure of the hip joint, the overall complication rate was 6% (six of ninety-six) and the reoperation rate was 3% (three of ninety-six). The rate of complications associated with the two-incision technique decreased significantly as the surgeon gained experience with the procedure (p = 0.0202). CONCLUSIONS: Although total hip arthroplasty with use of the two-incision technique was performed by a surgeon who was experienced in the performance of total hip replacement surgery with use of a single small incision, the rates of complications and repeat surgery associated with the two-incision technique initially were very high. While the rate diminished with increasing experience, total hip replacement with use of two incisions and fluoroscopic guidance is a technically demanding procedure that may be associated, especially initially, with higher rates of complications and repeat surgery. PMID- 16264119 TI - Outcomes of total elbow arthroplasty for rheumatoid arthritis: comparative study of three implants. AB - BACKGROUND: As the English-language literature on prosthetic elbow arthroplasty contains only two comparative studies of implants in contemporary use, to our knowledge, comparisons of prosthetic performance is difficult. An improved knowledge of comparative outcomes would be valuable in guiding implant selection. METHODS: We identified three groups of consecutive patients who had undergone prosthetic elbow arthroplasty with the Souter-Strathclyde, Kudo, or Coonrad Morrey implant for the treatment of rheumatoid arthritis. There were thirty-three elbows in each group. All procedures were done by or under the supervision of one surgeon. Surviving patients in whom the elbow had not been revised were followed for a mean of sixty-one months after treatment with the Souter-Strathclyde implant, sixty-seven months after treatment with the Kudo implant, and sixty eight months after treatment with the Coonrad-Morrey implant. Clinical function was assessed on the basis of pain relief and the range of flexion. Survivorship was assessed with use of a life-table method, with revision surgery and radiographic signs of loosening as the end points. RESULTS: The groups were comparable in terms of age, sex, and mean duration of follow-up. All three implant procedures relieved pain. Sustained improvement in the range of flexion was comparable among the three groups, with no implant procedure dramatically changing the fixed flexion deformity and all three improving maximum flexion. Revision surgery was needed because of infection, dislocation, and aseptic loosening. Survival of the Coonrad-Morrey implant was better than that of the other two implants. The five-year survival rates, with revision and radiographic signs of loosening as the end points, were 85% and 81% for the Souter-Strathclyde implant, 93% and 82% for the Kudo implant, and 90% and 86% for the Coonrad-Morrey implant. While radiographic evidence of loosening of the Coonrad-Morrey implants was less common, we noted focal osteolysis adjacent to 16% of these ulnar components and half of these cases progressed to frank loosening. CONCLUSIONS: The clinical function of these implants was similar in terms of pain relief and range of motion. We believe that component linkage with the Coonrad-Morrey implant prevents dislocation without increasing the risk of loosening. PMID- 16264120 TI - Use of distal femoral osteoarticular allografts in limb salvage surgery. AB - BACKGROUND: As diagnostic and therapeutic techniques improve, patients with a musculoskeletal sarcoma should expect longer survival, fewer complications and side effects, and an improved quality of life. Functional longevity of the reconstruction after resection of the tumor becomes a major concern, especially in young and physically active patients. The purpose of this study was to analyze the mid-term and long-term survival of reconstructions with a distal femoral osteoarticular allograft in a series of patients. METHODS: We retrospectively reviewed the results of eighty reconstructions with a distal femoral osteoarticular allograft following resection of a bone tumor in seventy-six patients. The mean duration of follow-up was eighty-two months. The rates of survival of the allograft and the joint surface were estimated with use of the Kaplan-Meier method. Cox regression analysis was performed to determine whether age, gender, the percentage of the femur that had been resected, and the use of chemotherapy were independent prognostic factors. Functional and radiographic results were documented according to the Musculoskeletal Tumor Society scoring system at the time of the latest follow-up. RESULTS: Five patients were lost to follow-up, leaving seventy-five allografts in seventy-one patients available for study. Thirteen patients (thirteen allografts) died of tumor-related causes without allograft failure before a two-year radiographic follow-up could be performed. Of the remaining sixty-two allografts, fourteen failed: six failed as a result of infection; four, because of local recurrence; one, because of massive resorption; and three, as a result of fracture. At the time of final follow-up, at a mean of 125 months, forty-eight allografts were still in place. The overall rate of allograft survival was 78% at both five and ten years, and the rate of allograft survival without the need for resurfacing with a knee prosthesis was 71% at both five and ten years. With the numbers available, age, gender, the percentage of the femur that had been resected, and the use of chemotherapy were not found to have a significant effect on the overall allograft survival rates. The patients who retained the original allograft had good or excellent functional and radiographic results. CONCLUSIONS: The life expectancy for most patients with a highly aggressive or malignant tumor in the distal part of the femur is now several decades. In this study, we found a high rate of survival of distal femoral allograft reconstructions at both five and ten years. PMID- 16264121 TI - Effect of femoral head diameter and operative approach on risk of dislocation after primary total hip arthroplasty. AB - BACKGROUND: It has been postulated that use of a larger femoral head could reduce the risk of dislocation after total hip arthroplasty, but only limited clinical data have been presented as proof of this hypothesis. METHODS: From 1969 to 1999, 21,047 primary total hip arthroplasties with varying femoral head sizes were performed at one institution. Patients routinely were followed at defined intervals and were specifically queried about dislocation. The operative approach was anterolateral in 9155 arthroplasties, posterolateral in 3646, and transtrochanteric in 8246. The femoral head diameter was 22 mm in 8691 of the procedures, 28 mm in 8797, and 32 mm in 3559. RESULTS: One or more dislocations occurred in 868 of the 21,047 hips. The cumulative risk of first-time dislocation was 2.2% at one year, 3.0% at five years, 3.8% at ten years, and 6.0% at twenty years. The cumulative ten-year rate of dislocation was 3.1% following anterolateral approaches, 3.4% following transtrochanteric approaches, and 6.9% following posterolateral approaches. The cumulative ten-year rate of dislocation was 3.8% for 22-mm-diameter femoral heads, 3.0% for 28-mm heads, and 2.4% for 32 mm heads in hips treated with an anterolateral approach; 3.5% for 22-mm heads, 3.5% for 28-mm heads, and 2.8% for 32-mm heads in hips treated with a transtrochanteric approach; and 12.1% for 22-mm heads, 6.9% for 28-mm heads, and 3.8% for 32-mm heads in hips treated with a posterolateral approach. Multivariate analysis showed the relative risk of dislocation to be 1.7 for 22-mm compared with 32-mm heads and 1.3 for 28-mm compared with 32-mm heads. CONCLUSIONS: In total hip arthroplasty, a larger femoral head diameter was associated with a lower long-term cumulative risk of dislocation. The femoral head diameter had an effect in association with all operative approaches, but the effect was greatest in association with the posterolateral approach. PMID- 16264122 TI - The accuracy of diagnostic imaging for the assessment of chronic osteomyelitis: a systematic review and meta-analysis. AB - BACKGROUND: A variety of diagnostic imaging techniques is available for excluding or confirming chronic osteomyelitis. Until now, an evidence-based algorithmic model for choosing the most suitable imaging technique has been lacking. The objective of this study was to determine the accuracy of current imaging modalities in the diagnosis of chronic osteomyelitis. METHODS: A systematic review and meta-analysis of the literature was conducted with a comprehensive search of the MEDLINE, EMBASE, and Current Contents databases to identify clinical studies on chronic osteomyelitis that evaluated diagnostic imaging modalities. The value of each imaging technique was studied by determining its sensitivity and specificity compared with the results of histological analysis, findings on culture, and clinical follow-up of more than six months. RESULTS: A total of twenty-three clinical studies in which the accuracy was described for radiography (two studies), magnetic resonance imaging (five), computed tomography (one), bone scintigraphy (seven), leukocyte scintigraphy (thirteen), gallium scintigraphy (one), combined bone and leukocyte scintigraphy (six), combined bone and gallium scintigraphy (three), and fluorodeoxyglucose positron emission tomography (four) were included in the review. No meta-analysis was performed with respect to computed tomography, gallium scintigraphy, and radiography. Pooled sensitivity demonstrated that fluorodeoxyglucose positron emission tomography was the most sensitive technique, with a sensitivity of 96% (95% confidence interval, 88% to 99%) compared with 82% (95% confidence interval, 70% to 89%) for bone scintigraphy, 61% (95% confidence interval, 43% to 76%) for leukocyte scintigraphy, 78% (95% confidence interval, 72% to 83%) for combined bone and leukocyte scintigraphy, and 84% (95% confidence interval, 69% to 92%) for magnetic resonance imaging. Pooled specificity demonstrated that bone scintigraphy had the lowest specificity, with a specificity of 25% (95% confidence interval, 16% to 36%) compared with 60% (95% confidence interval, 38% to 78%) for magnetic resonance imaging, 77% (95% confidence interval, 63% to 87%) for leukocyte scintigraphy, 84% (95% confidence interval, 75% to 90%) for combined bone and leukocyte scintigraphy, and 91% (95% confidence interval, 81% to 95%) for fluorodeoxyglucose positron emission tomography. The sensitivity of leukocyte scintigraphy in detecting chronic osteomyelitis in the peripheral skeleton was 84% (95% confidence interval, 72% to 91%) compared with 21% (95% confidence interval, 11% to 38%) for its detection of chronic osteomyelitis in the axial skeleton. The specificity of leukocyte scintigraphy in the axial skeleton was 60% (95% confidence interval, 39% to 78%) compared with 80% (95% confidence interval, 61% to 91%) for the peripheral skeleton. CONCLUSIONS: Fluorodeoxyglucose positron emission tomography has the highest diagnostic accuracy for confirming or excluding the diagnosis of chronic osteomyelitis. Leukocyte scintigraphy has an appropriate diagnostic accuracy in the peripheral skeleton, but fluorodeoxyglucose positron emission tomography is superior for detecting chronic osteomyelitis in the axial skeleton. PMID- 16264123 TI - Economic evaluation of ultrasonography in the diagnosis and management of developmental hip dysplasia in the United Kingdom and Ireland. AB - BACKGROUND: Clinical neonatal hip screening is performed to identify hip instability and the increased risk of later hip subluxation and dislocation. However, there is minimal information regarding the costs of such screening to parents and health services. The aim of this study was to assess these costs in association with the use of ultrasonography for the diagnosis and management of neonatal hip instability. METHODS: We conducted a prospective economic analysis in conjunction with a randomized clinical trial (the Hip Trial), for which 629 patients were recruited from thirty-three centers in the United Kingdom and Ireland to be randomized to undergo either ultrasonographic hip examination (314 patients) or clinical assessment alone (315 patients). Information on clinical outcomes was obtained from hospital records and records from the Hip Trial. Resource information was obtained from hospital records and from repeated periodic cross-sectional surveys of the families. Typical unit costs were applied to resource information to obtain a cost per patient, and the mean costs in the two study groups were calculated and compared. RESULTS: The average overall health-service cost per patient (and standard deviation) was $1298 +/- $2168 in the ultrasonography group and $1488 +/- $2912 in the group that underwent clinical assessment alone, a net difference of -$190 (95% confidence interval, $630 to $250). Families in which the infant was examined with ultrasonography had significantly lower costs associated with splinting: $92 compared with $118 in the group that underwent clinical assessment alone, a mean difference of -$26 (95% confidence interval, -$46 to -$6). Costs associated with surgery and total costs to the family were also slightly, but not significantly, lower in the ultrasonography group. CONCLUSIONS: Our results suggest that use of ultrasonography in the management of neonates with clinical hip instability is unlikely to impose an increased cost burden and may reduce costs to health services and families. PMID- 16264124 TI - Traumatic atlanto-occipital dislocation in children. AB - BACKGROUND: Traumatic atlanto-occipital dislocation in children and adolescents is a rare and often fatal injury. Although historically most reported cases have been fatal, the advent of modern prehospital care has led to an increase in survival following this injury. As a consequence, some patients may achieve or maintain satisfactory neurologic function following early intervention, stabilization, and definitive management. We analyzed the data on children and adolescents in whom traumatic atlanto-occipital dislocation had been treated with modern resuscitation techniques at our institution. METHODS: Atlanto-occipital dislocation is defined as disruption of the ligaments and other supporting soft tissues as indicated by displacement in either a transverse or vertical direction. With use of the Trauma Registry database at our institution, we identified sixteen such injuries that had occurred between 1986 and 2003. The hospital charts, clinic notes, and radiographs were reviewed. A careful neurological evaluation was performed for all of the survivors at the time of the latest follow-up. RESULTS: The mean age of the sixteen patients at the time of the injury was 7.6 years. The mechanisms of injury were diverse. The mean Glasgow Coma Scale score was 7.4 points. Eleven of the sixteen patients underwent intubation in the field, two were intubated in the emergency department, and three were not intubated. Eight of the sixteen patients were declared dead on arrival in the emergency department. The eight surviving patients initially were immobilized with either a halo vest or another orthosis. All patients except one received intravenous steroids in the emergency department. Three of the patients who survived the initial injury subsequently died while undergoing neurosurgical procedures for the treatment of extensive intracranial injuries. Four of the remaining five survivors underwent occiput-C2 fusion, and one was managed with a Minerva cast. At the time of the final follow-up, at a mean of 4.2 years after the injury, one patient was neurologically normal, three had mild spastic hemiparesis and were very functional, and one had spastic quadriplegia and was ventilator-dependent. CONCLUSIONS: Prompt recognition and treatment of traumatic atlanto-occipital dislocation in children and adolescents can result in improved survival. Early diagnosis, prompt intubation, early and adequate immobilization of the head and neck, and the use of intravenous steroids appear to facilitate survival. We recommend arthrodesis from the occiput to C2 (or the nearest adjacent intact and stable vertebra caudad to C2) for all children who survive a traumatic atlanto-occipital dislocation, particularly those with an incomplete spinal cord injury. PMID- 16264125 TI - A highly sensitive polymerase chain reaction method detects activating mutations of the GNAS gene in peripheral blood cells in McCune-Albright syndrome or isolated fibrous dysplasia. AB - BACKGROUND: The somatic nature of mutations in the GNAS gene in McCune-Albright syndrome and isolated fibrous dysplasia makes their identification difficult. Conventional methods for the detection of mosaic mutations of GNAS have required polymerase chain reaction analysis of genomic DNA from affected tissues or multiple rounds of tandem polymerase chain reaction and endonuclease digestion to enrich for mutant alleles in genomic deoxyribonucleic acid (DNA) from other tissues. Peptide nucleic acid (PNA) primers specifically block synthesis from the nonmutant or wild-type allele. We therefore used PNA-clamping to detect low copy numbers of mutant GNAS alleles in DNA from peripheral blood cells from patients with McCune-Albright syndrome and fibrous dysplasia. METHODS: We applied the PNA clamping method to the analysis of genomic DNA from peripheral blood cells of thirteen patients with McCune-Albright syndrome and three patients with isolated fibrous dysplasia. Polymerase chain reaction was performed in the presence and absence of PNA, and the polymerase chain reaction products were sequenced. In the absence of PNA, a strong 325 base-pair polymerase chain reaction band was generated from all samples; in the presence of PNA, there was an approximately 50% to 90% reduction in the intensity of this polymerase chain reaction product. RESULTS: In the absence of PNA, direct sequencing of the polymerase chain reaction products demonstrated R201 mutations in GNAS alleles of three of the thirteen patients with McCune-Albright syndrome and none of the three patients with fibrous dysplasia. In contrast, in the presence of PNA, R201 mutations were detected in eleven of the thirteen patients with McCune-Albright syndrome and in all three of the patients with fibrous dysplasia. In mixing experiments involving the use of wild-type and mutant DNA samples, we were able to determine the presence of a mutant GNAS allele in the equivalent of one cell in 1000 to 5000 cells. CONCLUSIONS: Inclusion of a specific PNA primer in the polymerase chain reaction for GNAS exon 8 allows the selective amplification of low numbers of mutant alleles, and it permits detection of activating mutations in genomic DNA from peripheral blood cells in patients with McCune-Albright syndrome and fibrous dysplasia. PMID- 16264126 TI - The ponticulus posticus: implications for screw insertion into the first cervical lateral mass. AB - BACKGROUND: The arcuate foramen is an important osseous anomaly of the first cervical vertebra (the atlas) that must be taken into consideration during placement of lateral mass screws into the atlas. METHODS: The prevalence of this anomaly in our patient population was determined through a retrospective review of 464 lateral radiographs of the neck. The anatomy of the arcuate foramen was identified in a study of cadavers. RESULTS: Seventy-two arcuate foramina, complete or incomplete, were identified on the 464 lateral radiographs of the neck; thus, the prevalence was 15.5% in our patient population. CONCLUSIONS: Although the arcuate foramen is a common anomaly, it is often not recognized. Proper identification of this anomaly on preoperative lateral radiographs should alert the surgeon to avoid using the ponticulus posticus as a starting point for a lateral mass screw in order to not injure the vertebral artery. PMID- 16264127 TI - Femoral component revision with use of impaction bone-grafting and a cemented polished stem. AB - BACKGROUND: The purpose of this study was to evaluate the clinical and radiographic outcomes of revision of the femoral component of a hip arthroplasty with use of an impaction bone-grafting technique and a cemented polished stem. METHODS: Thirty-three consecutive femoral reconstructions that were performed between March 1991 and February 1996 with use of the X-change femoral revision system, fresh-frozen morselized allograft, and a cemented polished Exeter stem were followed prospectively. Femoral bone stock defects were classified according to the Endoklinik classification. The average age of the patients at the time of the femoral component revision was sixty-three years. No patient was lost to follow-up, which was performed at a minimum of eight years, but eight patients had died. None of the deaths was related to the surgery. RESULTS: No femoral reconstruction had been rerevised at a mean of 10.4 years postoperatively. There was one unrecognized intraoperative fracture, which healed following nonoperative treatment. There were three postoperative femoral fractures, all through cortical defects at the level of the tip of the prostheses. All fractures healed after plate fixation, and all femoral implants were left in situ. The average subsidence of the stem within the cement mantle was 3 mm; seven stems migrated > or =5 mm. The average Harris hip score improved from 49 points prior to surgery to 85 points (range, 68 to 100 points) at the time of this review. Subsidence did not affect the Harris hip score. Kaplan-Meier analysis, with an end point of femoral revision for any reason, showed a survival rate of 100% (one-sided 95% confidence interval, 100% to 91.3%). CONCLUSIONS: Femoral revision with use of an impaction bone-grafting technique and a cemented polished stem resulted in an excellent prosthetic survival rate at eight to thirteen years postoperatively. The major problem that occurred was a femoral fracture in four patients. PMID- 16264128 TI - Strain on the ulnar nerve at the elbow and wrist during throwing motion. AB - BACKGROUND: It is well known that cubital tunnel syndrome frequently occurs in throwing athletes. The cause of cubital tunnel syndrome is considered to be mechanical stimuli on the ulnar nerve in the cubital tunnel. The hypothesis of the present cadaveric study was that the ulnar nerve is subjected to longitudinal strain in the cubital tunnel during the throwing motion. METHODS: Four phases of throwing (stance, wind-up, middle cock-up, and early acceleration) were passively simulated in seven fresh-frozen transthoracic cadaveric specimens that were fixed in an upright position to allow free arm movement. In each throwing phase, the elbow was sequentially flexed from 45 degrees to 90 degrees to 120 degrees to maximum flexion. The longitudinal movement of and strain on the ulnar nerve were measured with use of a caliper and a strain gauge at the proximal aspects of both the cubital tunnel and the canal of Guyon. RESULTS: The movement of the ulnar nerve at the proximal aspect of the cubital tunnel was significantly increased during all throwing phases with increased elbow flexion (p < 0.05). An average maximum movement of 12.4 +/- 2.4 mm was recorded during the wind-up phase with maximum elbow flexion. The movement at the proximal aspect of the canal of Guyon was approximately two-thirds of that at the proximal aspect of the cubital tunnel. The strain on the ulnar nerve at the proximal aspect of the cubital tunnel was significantly increased with elbow flexion in the stance, wind-up, and middle cock-up phases (p < 0.05). An average maximum strain of 13.1% +/- 6.1% was recorded during the early acceleration phase with maximum elbow flexion. The strain at the proximal aspect of the canal of Guyon was approximately half of that at the proximal aspect of the cubital tunnel. CONCLUSIONS: In the present study, the maximum strain on the ulnar nerve during the acceleration phase was found to be close to the elastic and circulatory limits of the nerve. PMID- 16264129 TI - Reamed femoral nailing in sheep: does irrigation and aspiration of intramedullary contents alter the systemic response? AB - BACKGROUND: Reaming of the femoral canal has been demonstrated to introduce intramedullary contents into the circulation with subsequent pulmonary embolization. The aim of this study was to investigate whether this effect can be minimized by use of a reamer system that provides simultaneous irrigation and aspiration of intramedullary contents. METHODS: A unilateral lung contusion was created and intramedullary femoral nailing was subsequently performed in eighteen female skeletally mature Merino sheep. The animals were divided into three groups, of six animals each, to receive one of three types of treatment: reamed femoral nailing; reaming, irrigation, and aspiration; and unreamed femoral nailing. Blood samples were obtained and a bronchoalveolar lavage was performed at baseline, immediately after creation of the lung contusion, immediately after intramedullary nailing, and at four hours after surgery. Pulmonary permeability, polymorphonuclear leukocyte activity, and systemic hemostatic response were measured. Lung specimens were obtained for histological evaluation. RESULTS: At baseline and immediately after creation of the lung contusion, endothelial permeability was comparable among the three groups. At four hours postoperatively, pulmonary permeability was significantly higher in the group treated with reamed femoral nailing (urea/protein ratio; 256.7) than in the group treated with reaming, irrigation, and aspiration (urea/protein ratio, 91.5) and the group treated with unreamed femoral nailing (urea/protein, 110.64) (p < 0.05). The stimulatory capacity of the polymorphonuclear leukocytes was significantly decreased (p < 0.05) only in the group treated with reamed femoral nailing; the other two groups had no significant decrease postoperatively (p > 0.05). The D-dimer level at four hours postoperatively was significantly higher in the group treated with reamed femoral nailing than it was in the other two groups (p < 0.05). Histological examination showed that the grades of edema and polymorphonuclear leukocyte diapedesis were also highest in the group treated with reamed femoral nailing. CONCLUSIONS: It appears that, in the presence of a unilateral pulmonary injury, the systemic effects of intramedullary reaming of an intact femur can be minimized with use of a modified reamer design that simultaneously irrigates the canal and removes debris. Additional clinical validation of this reaming system is necessary. PMID- 16264130 TI - Congenital anomalies of the ribs and chest wall associated with congenital deformities of the spine. AB - BACKGROUND: Congenital anomalies of the ribs and chest wall as well as Sprengel deformity of the shoulder are often associated with congenital deformities of the spine. It has been suggested that rib anomalies may adversely affect the progression of the spinal deformity. METHODS: We conducted a retrospective study of the medical records and spine radiographs of 620 consecutive patients with congenital deformities of the spine; 497 patients (80%) had scoliosis; eighty eight patients (14%), kyphoscoliosis; and thirty-five patients (6%), kyphosis. The rib anomalies were classified into simple and complex, and the presence of a Sprengel deformity of the shoulder was recorded. The rate of scoliosis deterioration without treatment before the age of eleven years, as well as the patient age and curve size at the time of surgery, was compared for different types of vertebral abnormalities in patients with and without rib anomalies. RESULTS: A total of 119 patients (19.2%) had rib anomalies, which were most commonly associated with congenital scoliosis (111 patients; 93%) and were much less frequently associated with congenital kyphoscoliosis or kyphosis (eight patients). The rib anomalies were simple in ninety-five patients and complex in twenty-four. Eighty-five patients (71%) with rib abnormalities had a scoliosis due to a unilateral failure of vertebral segmentation, and seven patients had mixed or unclassifiable vertebral anomalies. In contrast, only sixteen of 203 patients with a scoliosis due to a hemivertebra alone had rib anomalies. The rib anomalies were most frequently associated with a thoracic or thoracolumbar scoliosis (102 patients; 92%) and occurred on the concavity in eighty-two patients (74%), the convexity in twenty-two patients (20%), and were bilateral in seven patients. The Sprengel deformity occurred in forty-five patients and most frequently in association with a thoracic scoliosis due to a unilateral failure of vertebral segmentation (twenty-seven patients). No significant difference was detected in the rate of curve progression without treatment in patients with and without rib anomalies. The only exception was the mean age at the time of surgery, which was higher for patients with a unilateral unsegmented bar without rib anomalies (p = 0.005). In addition, no significant difference was found with regard to any tethering effect due to the site of the rib fusions on the concavity of the scoliosis, i.e., whether they were in close approximation to the spine or were more lateral (p > 0.05). CONCLUSIONS: Congenital rib anomalies occur most commonly on the concavity of a thoracic or thoracolumbar congenital scoliosis that is due to a unilateral failure of vertebral segmentation, and they do not appear to have an adverse effect on curve size or rate of progression. PMID- 16264131 TI - Acute torticollis after isolated stress fracture of the first rib in a child. A case report. PMID- 16264133 TI - Hemophilic pseudotumor of the distal parts of the radius and ulna. A case report. PMID- 16264132 TI - Bilateral gluteal compartment syndrome. A case report. PMID- 16264134 TI - Molecular biology in orthopaedics: the advent of molecular orthopaedics. AB - Molecular biology is the study, at the molecular level, of how genetic information is stored, inherited, and expressed and how it influences the structure and function of cells. Although molecular biology approaches have been used for decades in orthopaedic research, they are only now beginning to influence clinical practice. A variety of sophisticated techniques permit rapid and affordable DNA sequencing, gene expression profiling, gene cloning, gene manipulation, gene transfer, recombinant protein production, and other technologies of enormous biomedical importance. Success in genomics has spawned additional ambitious endeavors, including proteomics, pharmacogenetics, and bioinformatics. These techniques are providing new diagnostic, staging, prognostic, and therapeutic opportunities in all areas of medicine, including orthopaedics. With the use of molecular criteria, treatment of the orthopaedic patient may become more individualized, and greater emphasis will be placed on preventative strategies based on the patient's genetic makeup. Both surgical and nonsurgical decisions will increasingly accommodate molecular criteria. PMID- 16264135 TI - Symposium resident work-hour guidelines. A sentence or an opportunity for orthopaedic education? PMID- 16264137 TI - Comparison of soap and antibiotic solutions for irrigation of lower-limb open fracture wounds. PMID- 16264138 TI - Anterior cruciate ligament reconstruction with a four-strand hamstring tendon autograft. PMID- 16264136 TI - The role of patient restrictions in reducing the prevalence of early dislocation following total hip arthroplasty. PMID- 16264139 TI - Effect of bisphosphonates on periprosthetic bone mineral density after total joint arthroplasty. PMID- 16264140 TI - Effect of humeral component anteversion on shoulder stability with glenoid component retroversion. PMID- 16264141 TI - Device for zone-II flexor tendon repair. PMID- 16264142 TI - Rate of improvement was not different after osteochondral repair with matrix induced autologous chondrocyte implantation or autologous chondrocyte implantation with a cover made from porcine-derived type I/type III collagen. PMID- 16264143 TI - Continuous passive motion improves active knee flexion and shortens hospital stay but does not affect other functional outcomes after knee arthroplasty. PMID- 16264144 TI - Arthroscopic decompression and physiotherapy have similar effectiveness for subacromial impingement. PMID- 16264146 TI - The cover. Laid Off. PMID- 16264147 TI - A piece of my mind. Playing doctor. PMID- 16264148 TI - Safety improvements urged for MRI facilities. PMID- 16264149 TI - Experts ponder pediatric research ethics. PMID- 16264150 TI - New mind/body tactics target medically unexplained physical symptoms and fears. PMID- 16264151 TI - Children's health study closer to launch: lack of funding could cause delays. PMID- 16264152 TI - Fish oil supplementation and arrhythmias. PMID- 16264153 TI - Life-threatening complications from doxorubicin-docetaxel chemotherapy for breast cancer. PMID- 16264154 TI - Self-reported sexual function in women and androgen levels. PMID- 16264156 TI - Earthquakes and Takotsubo cardiomyopathy. PMID- 16264155 TI - Long-term outcomes for extremely low-birth-weight infants. PMID- 16264157 TI - Parvovirus B19 and onset of juvenile dermatomyositis. PMID- 16264158 TI - Screen-and-treat approaches for cervical cancer prevention in low-resource settings: a randomized controlled trial. AB - CONTEXT: Non-cytology-based screen-and-treat approaches for cervical cancer prevention have been developed for low-resource settings, but few have directly addressed efficacy. OBJECTIVE: To determine the safety and efficacy of 2 screen and-treat approaches for cervical cancer prevention that were designed to be more resource-appropriate than conventional cytology-based screening programs. DESIGN, SETTING, AND PATIENTS: Randomized clinical trial of 6555 nonpregnant women, aged 35 to 65 years, recruited through community outreach and conducted between June 2000 and December 2002 at ambulatory women's health clinics in Khayelitsha, South Africa. INTERVENTIONS: All patients were screened using human papillomavirus (HPV) DNA testing and visual inspection with acetic acid (VIA). Women were subsequently randomized to 1 of 3 groups: cryotherapy if she had a positive HPV DNA test result; cryotherapy if she had a positive VIA test result; or to delayed evaluation. MAIN OUTCOME MEASURES: Biopsy-confirmed high-grade cervical cancer precursor lesions and cancer at 6 and 12 months in the HPV DNA and VIA groups compared with the delayed evaluation (control) group; complications after cryotherapy. RESULTS: The prevalence of high-grade cervical intraepithelial neoplasia and cancer (CIN 2+) was significantly lower in the 2 screen-and-treat groups at 6 months after randomization than in the delayed evaluation group. At 6 months, CIN 2+ was diagnosed in 0.80% (95% confidence interval [CI], 0.40%-1.20%) of the women in the HPV DNA group and 2.23% (95% CI, 1.57%-2.89%) in the VIA group compared with 3.55% (95% CI, 2.71%-4.39%) in the delayed evaluation group (P<.001 and P = .02 for the HPV DNA and VIA groups, respectively). A subset of women underwent a second colposcopy 12 months after enrollment. At 12 months the cumulative detection of CIN 2+ among women in the HPV DNA group was 1.42% (95% CI, 0.88%-1.97%), 2.91% (95% CI, 2.12%-3.69%) in the VIA group, and 5.41% (95% CI, 4.32%-6.50%) in the delayed evaluation group. Although minor complaints, such as discharge and bleeding, were common after cryotherapy, major complications were rare. CONCLUSION: Both screen-and-treat approaches are safe and result in a lower prevalence of high-grade cervical cancer precursor lesions compared with delayed evaluation at both 6 and 12 months. Trial Registration http://clinicaltrials.gov Identifier: NCT00233727. PMID- 16264159 TI - Feasibility of management of high-grade cervical lesions in a single visit: a randomized controlled trial. AB - CONTEXT: The incidence of cervical cancer is higher among low-income and minority women who have never undergone a conventional Papanicolaou test or who do not follow up after testing. Screening has been shown to reduce cervical cancer incidence rates. OBJECTIVES: To determine the feasibility and acceptability of immediately treating women with severely abnormal Papanicolaou test results by using a single-visit cervical cancer screening and treatment program and to compare treatment rates and 12-month follow-up rates with those of women who received usual care. DESIGN, SETTING, AND PARTICIPANTS: Randomized controlled trial conducted among 3521 women aged 18 years or older recruited between January 1999 and April 2002 at US community health centers located in predominantly Latino underserved areas. INTERVENTIONS: Women randomized to usual care (n = 1805) were discharged immediately after examination. Women randomized to the single-visit group (n = 1716) remained at the clinic until the results of their conventional Papanicolaou test were available. Large loop electrosurgical excision procedure was performed in single-visit patients with either a diagnosis of a high-grade squamous intraepithelial lesion (HGSIL)/atypical glandular cells of undetermined significance (AGUS) or suspicion of carcinoma. All other patients with abnormal Papanicolaou test results were referred to abnormal cytology clinics or elected to receive follow-up care outside the study's medical system. MAIN OUTCOME MEASURES: Treatment rates for HGSIL/AGUS at 6 months, follow-up rates at 6 months for lower-grade lesions, and 1-year follow-up rates for all patients. RESULTS: The rate of abnormal Papanicolaou test results was 4.1%. One percent of results showed high-grade lesions. In the single-visit group, the mean visit time was 2.8 hours and the mean time for delivery and processing of the Papanicolaou tests was 66 minutes. Six months after randomization, 14 (88%) of 16 single-visit and 10 (53%) of 19 usual care patients with HGSIL/AGUS had completed treatment. Fifty percent in the single-visit program and 53% of usual care with less abnormal Papanicolaou tests completed treatment within 6 months. Overall, 36% in each group presented for a follow-up Papanicolaou test 1 year later. Women in the single-visit group with high-grade lesions (10/16; 63%) were significantly more likely to attend follow-up for Papanicolaou tests 12 months later than women with similar lesions in the usual care group (4/19; 21%). CONCLUSIONS: For cervical cancer screening, the single-visit program was feasible and the degree of acceptability was high in this underserved population. Single visit programs provide an opportunity to increase the rate of immediate treatment and follow-up of women with severely abnormal Papanicolaou test results. This strategy did not improve follow-up rates for women with less-abnormal results. Trial Registration http://ClinicalTrials.govIdentifier: NCT00237562. PMID- 16264160 TI - Neurological and neuromuscular disease as a risk factor for respiratory failure in children hospitalized with influenza infection. AB - CONTEXT: The Advisory Committee on Immunization Practices (ACIP) recommends annual influenza vaccination for children with certain chronic medical conditions to prevent serious complications of influenza infection. Little is known about the relative contribution of each of these chronic medical conditions to the development of serious influenza-associated complications. OBJECTIVE: To identify chronic medical conditions that are associated with respiratory failure in children hospitalized with community-acquired laboratory-confirmed influenza. DESIGN, SETTING, AND PATIENTS: A retrospective cohort study of patients aged 21 years or younger hospitalized at The Children's Hospital of Philadelphia with community-acquired laboratory-confirmed influenza during 4 consecutive influenza seasons (June 2000 through May 2004). We examined 9 ACIP-designated high-risk chronic medical conditions and 3 additional chronic medical conditions (neurological and neuromuscular disease [NNMD], gastroesophageal reflux disease [GERD], and history of prematurity) that in recent studies have been associated with influenza hospitalization and severe influenza-related complications. MAIN OUTCOME MEASURES: Rate and odds ratio (OR) of respiratory failure, defined as need for mechanical ventilation. RESULTS: Of 745 children hospitalized with community-acquired laboratory-confirmed influenza, 322 (43%) had 1 or more ACIP designated high-risk chronic medical conditions. Neurological and neuromuscular disease, GERD, and history of prematurity were present in 12%, 14%, and 3%, of children, respectively. Thirty-two children (4.3%) developed respiratory failure. In multivariate logistic regression analyses, conditions associated with respiratory failure included NNMD (OR, 6.0; 95% confidence interval [CI], 2.7 13.5), chronic pulmonary disease other than asthma (OR, 4.8; 95% CI, 1.5-15.1), and cardiac disease (OR, 4.0; 95% CI, 1.6-10.2). The predicted probabilities of respiratory failure derived from the multivariate model were 12% (95% CI, 7% 20%), 9% (95% CI, 3%-23%), and 8% (95% CI, 4%-18%) for children with NNMD, chronic pulmonary disease, and cardiac disease, respectively. CONCLUSIONS: These results support the ACIP's recent decision to add NNMD to the list of conditions for which annual influenza vaccine is recommended in children. Neurologists and primary care pediatricians should be alerted to the increased risk of respiratory failure and the importance of influenza vaccination in children with NNMD. PMID- 16264161 TI - History and molecular genetics of Lynch syndrome in family G: a century later. AB - CONTEXT: In 1895, Aldred Scott Warthin, MD, PhD, initiated one of the most thoroughly documented and longest cancer family histories ever recorded. The unusually high incidence and segregation of cancers of the colon, rectum, stomach, and endometrium in Dr Warthin's family G was later followed up by his colleagues, most recently by Henry Lynch, MD. Described today as a Lynch syndrome family, family G was last documented in 1971, prior to the modern era of molecular diagnostics. OBJECTIVE: To update family G. DESIGN, SETTING, AND PARTICIPANTS: Historical prospective cohort study of family G members from 1895 to 2000. MAIN OUTCOME MEASURES: The primary outcomes were the frequencies and types of cancers, ages at diagnosis, and presence of the T to G transversion at the splice acceptor site of exon 4 of the mutS homolog 2, colon cancer, nonpolyposis type 1 (E coli) (MSH2) gene in family G members. A secondary analysis compared cancer-specific incidence rates in family G with published national and regional cancer incidence rates through the standardized incidence ratio (SIR). RESULTS: Family G now has 929 known descendants of the original progenitor first reported in 1913. Cancers of the colon and rectum (SIR, 3.20; 95% confidence interval [CI], 2.39-4.19) and endometrium (SIR, 3.51; 95% CI, 1.92 5.89) continue to predominate in family G. Five of 40 tested members of family G carry the MSH2 T to G mutation; as a result, 15 of their living relatives are at increased risk of developing 1 or more colorectal or Lynch syndrome-associated cancers. In contrast, 97 living members of family G can now be excluded as mutation carriers. CONCLUSION: Within the last decade, molecular diagnostic testing has transformed the care of family G and other Lynch syndrome families in which a pathogenic mutation has been identified. PMID- 16264163 TI - A 21-year-old woman with atypical squamous cells of undetermined significance. PMID- 16264162 TI - Randomized trials stopped early for benefit: a systematic review. AB - CONTEXT: Randomized clinical trials (RCTs) that stop earlier than planned because of apparent benefit often receive great attention and affect clinical practice. Their prevalence, the magnitude and plausibility of their treatment effects, and the extent to which they report information about how investigators decided to stop early are, however, unknown. OBJECTIVE: To evaluate the epidemiology and reporting quality of RCTs involving interventions stopped early for benefit. DATA SOURCES: Systematic review up to November 2004 of MEDLINE, EMBASE, Current Contents, and full-text journal content databases to identify RCTs stopped early for benefit. STUDY SELECTION: Randomized clinical trials of any intervention reported as having stopped early because of results favoring the intervention. There were no exclusion criteria. DATA EXTRACTION: Twelve reviewers working independently and in duplicate abstracted data on content area and type of intervention tested, reporting of funding, type of end point driving study termination, treatment effect, length of follow-up, estimated sample size and total sample studied, role of a data and safety monitoring board in stopping the study, number of interim analyses planned and conducted, and existence and type of monitoring methods, statistical boundaries, and adjustment procedures for interim analyses and early stopping. DATA SYNTHESIS: Of 143 RCTs stopped early for benefit, the majority (92) were published in 5 high-impact medical journals. Typically, these were industry-funded drug trials in cardiology, cancer, and human immunodeficiency virus/AIDS. The proportion of all RCTs published in high impact journals that were stopped early for benefit increased from 0.5% in 1990 1994 to 1.2% in 2000-2004 (P<.001 for trend). On average, RCTs recruited 63% (SD, 25%) of the planned sample and stopped after a median of 13 (interquartile range [IQR], 3-25) months of follow-up, 1 interim analysis, and when a median of 66 (IQR, 23-195) patients had experienced the end point driving study termination (event). The median risk ratio among truncated RCTs was 0.53 (IQR, 0.28-0.66). One hundred thirty-five (94%) of the 143 RCTs did not report at least 1 of the following: the planned sample size (n = 28), the interim analysis after which the trial was stopped (n = 45), whether a stopping rule informed the decision (n = 48), or an adjusted analysis accounting for interim monitoring and truncation (n = 129). Trials with fewer events yielded greater treatment effects (odds ratio, 28; 95% confidence interval, 11-73). CONCLUSIONS: RCTs stopped early for benefit are becoming more common, often fail to adequately report relevant information about the decision to stop early, and show implausibly large treatment effects, particularly when the number of events is small. These findings suggest clinicians should view the results of such trials with skepticism. PMID- 16264164 TI - Role of the urban academic medical center in US health care. PMID- 16264165 TI - Psychiatry as a clinical neuroscience discipline. PMID- 16264166 TI - Cervical cancer prevention: making programs more appropriate and pragmatic. PMID- 16264167 TI - When (not) to stop a clinical trial for benefit. PMID- 16264168 TI - JAMA patient page. Randomized controlled trials. PMID- 16264169 TI - Risks of cancer and families. PMID- 16264170 TI - Doctor ... will I still be able to have children? PMID- 16264171 TI - Goal of maintaining public's trust brings research groups together on conflict-of interest guidelines. PMID- 16264172 TI - Sharks do get cancer: few surprises in cartilage research. PMID- 16264173 TI - Studies seek molecular clues on alcohol's role in cancer. PMID- 16264174 TI - Animal rights violence spreads fear through U.K. research community. PMID- 16264175 TI - Stat bite: Annual percent change in death rates for selected cancers,1992-2002. PMID- 16264176 TI - Reflections on findings of the Cancer Outcomes Measurement Working Group: moving to the next phase. AB - The Cancer Outcomes Measurement Working Group (COMWG) was a National Cancer Institute working group of 35 experts convened to examine the state of the science and identify future research priorities for outcomes assessment in cancer. The COMWG focused on three outcomes (health-related quality of life [HRQOL]), patient needs and satisfaction, and economic burden) in four cancers (breast, colorectal, lung, and prostate) across the continuum of care (prevention and screening, treatment, survivorship, and end of life). The majority of the research to date has focused on HRQOL assessment, which has been shown to be feasible in a research context, using questionnaires that meet established criteria for reliability and validity. The quality and quantity of HRQOL research has increased markedly in recent years, and additional methodological developments--particularly the application of item response theory to improve precision, efficiency, and comparability in measurement--hold considerable promise. Research is needed to develop and test predictive models of HRQOL and to establish the added value of including HRQOL assessment in clinical trials. PMID- 16264177 TI - Familial risk of cancer shortly after diagnosis of the first familial tumor. AB - BACKGROUND: The diagnosis of the first cancer in a family may lead to the medical examination of the patient's relatives and the subsequent identification of additional familial cancers. If detection bias is present, familial risks could be overestimated soon after first diagnosis. METHODS: We followed 1,677,722 offspring/siblings of 846,448 probands from the year of diagnosis of the first familial tumor to the diagnosis of first cancer, death, emigration, or December 31, 2002, using the Swedish Family Cancer Database. The risks of cancer among the offspring and siblings of patients with melanoma and cancers of the breast, prostate, colorectum, cervix, and lung were compared with those in the general population. Relative risks (RRs) were determined using Poisson regression, according to the time after first diagnosis. All statistical tests were two sided. RESULTS: Daughters of women with breast cancer had a statistically significantly higher relative risk of in situ breast cancer during the year of the mother's diagnosis than they did 5 or more years later (RR = 4.78, 95% confidence interval [CI] = 2.16 to 10.6, 26.6 cases per 100,000, versus RR = 1.97, 95% CI = 1.65 to 2.37, 27.2 cases per 100,000; P = .033). Daughters diagnosed the same year as their mothers were younger and were diagnosed earlier in the calendar year than daughters of women diagnosed 5 or more years after their mothers. Similarly, the risk of invasive melanoma among the offspring of individuals with invasive melanoma was higher during the year of the parent's diagnosis than it was 5 or more years afterward (RR = 8.27, 95% CI = 3.82 to 17.9, 57.0 cases per 100,000, versus RR = 3.18, 95% CI = 2.55 to 3.97, 37.6 cases per 100,000; P = .019). Sibling risks of in situ breast cancer, in situ cervical cancer, and invasive prostate cancer also decreased with time after diagnosis of the first familial tumor. CONCLUSIONS: Increased surveillance may result in the earlier detection of asymptomatic familial cancers, i.e., in detection bias. The possibility of overestimated familial risks of cancer shortly after diagnosis of the first familial tumor should be considered before a patient's clinical and genetic counseling is implemented. PMID- 16264178 TI - Paternity following treatment for testicular cancer. AB - BACKGROUND: Studies of fertility in men treated for testicular cancer have mainly addressed serum follicle-stimulating hormone levels and sperm parameters. We assessed post-treatment paternity among long-term survivors of testicular cancer. METHODS: Men (n = 1814) who had been treated for unilateral testicular cancer in Norway during 1980 through 1994 were invited to participate in a national multi center follow-up survey in 1998 through 2002. The participants were allocated to five groups according to the treatment received after orchiectomy, including treatment at relapse (surveillance, retroperitoneal lymph node dissection, radiotherapy, low-dose chemotherapy [i.e., < or = 850 mg cisplatin], and high dose chemotherapy [i.e., > 850 mg cisplatin]). Cox proportional hazards analysis was used to assess predictive factors for post-treatment paternity. Statistical tests were two-sided. RESULTS: A total of 1433 men were assessable, of whom 827 were fathers at diagnosis. Post-treatment conception was attempted by 554 men, among whom the overall 15-year actuarial post-treatment paternity rate was 71% (95% confidence interval [CI] = 66% to 75%) without the use of cryopreserved semen. This rate ranged from 48% (95% CI = 30% to 69%) in the high-dose chemotherapy group to 92% (95% CI = 78% to 98%) in the surveillance group (P < .001). The median actuarial time from diagnosis to the birth of the first child after treatment was 6.6 years overall but varied according to treatment. Assisted reproductive technologies were used by 22% of the couples who attempted conception after treatment. Dry ejaculation, treatment group, pretreatment fatherhood, and marital status were statistically significant independent predictors for post-treatment fatherhood, with dry ejaculation as the most important negative factor. CONCLUSIONS: Although the overall paternity rate after treatment for testicular cancer was high, the ability to conceive and the time to conception reflected the intensity of treatment. These data may help inform patients about their future ability to father biological children. PMID- 16264179 TI - Expression of transcription factor E2F1 and telomerase in glioblastomas: mechanistic linkage and prognostic significance. AB - BACKGROUND: Several tumor suppressor pathways have been identified as modulators of telomerase function. We examined the functional role of the retinoblastoma E2F1 pathway in regulating telomerase activity in malignant gliomas. METHODS: Adenovirus vectors were used to transfer cDNAs into human glioblastoma and sarcoma cells. Telomerase activity was assessed with a telomere repeat amplification protocol. Promoter activity in cancer cells was assessed with promoter-luciferase reporter constructs. Promoter binding was assessed with the chromatin immunoprecipitation (ChIP) assay. We isolated astrocytes from E2F1 transgenic mice and normal mice for in vivo studies. We evaluated the expression of E2F1 and hTERT (the catalytic subunit of human telomerase) mRNAs by reverse transcriptase-polymerase chain reaction and proteins in human glioblastoma samples by immunoblot analysis. Associations between survival among 61 glioblastoma multiforme patients and expression of E2F1 and hTERT mRNA and protein were examined with Kaplan-Meier analysis, the log-rank test, and Cox proportional hazards regression models. All statistical tests were two-sided. RESULTS: Ectopic E2F1 expression increased hTERT promoter activity in cancer cells. We detected an interaction between E2F1 protein and the hTERT promoter. Transgenic E2F1 astrocytes contained functional telomerase protein. E2F1 mRNA expression and hTERT mRNA expression were statistically significantly correlated in human glioblastoma specimens (R = .8; P < .001). Longer median survival was statistically significantly associated with lower E2F1 mRNA expression in tumors (103.6 weeks) rather than with higher expression (46.1 weeks) (difference = 57.5 weeks; 95% confidence interval [CI] = 14.7 to 159.7; log-rank P = .002). E2F1 mRNA was the only factor that was statistically significantly associated with overall survival in a multivariable model (P = .04). Among 27 patients with glioblastoma multiforme samples, the expression of E2F1 protein was statistically significantly associated with survival (log-rank P < .001). CONCLUSIONS: E2F1 may participate in telomerase activity regulation in malignant glioma cells. Its expression appears to be strongly associated with the survival of patients with malignant brain tumors. PMID- 16264180 TI - Alcohol and postmenopausal breast cancer risk defined by estrogen and progesterone receptor status: a prospective cohort study. AB - BACKGROUND: Alcohol intake has been reported to be positively associated with an increased risk of postmenopausal breast cancer; however, the association with the estrogen receptor (ER) and progesterone receptor (PR) status of the breast tumors remains unclear. METHODS: Self-reported data on alcohol consumption were collected in 1987 and 1997 from 51,847 postmenopausal women in the population based Swedish Mammography Cohort. Through June 30, 2004, 1188 invasive breast cancer case patients with known ER and PR status were identified during an average 8.3-year follow-up. We used Cox proportional hazards models to estimate multivariable relative risks (RRs) of breast cancer, adjusting for age; family history of breast cancer; body mass index; height; parity; age at menarche, first birth, and menopause; education level; use of postmenopausal hormones; and diet. Heterogeneity among groups was evaluated using the Wald test. All statistical tests were two-sided. RESULTS: Alcohol consumption was associated with an increased risk for the development of ER-positive (+) tumors, irrespective of PR status (highest intake [> or = 10 g of alcohol per day] versus nondrinkers, multivariable RR = 1.35, 95% confidence interval [CI] = 1.02 to 1.80; Ptrend < .049 for ER+PR+ tumors; and RR = 2.36, 95% CI = 1.56 to 3.56; Ptrend < .001 for ER+PR-tumors). The absolute rate of ER+ breast cancer (standardized to the age distribution of person-years experienced by all study participants using 5-year age categories) was 232 per 100,000 person-years among women in the highest category of alcohol intake, and 158 per 100,000 person-years among nondrinkers. No association was observed between alcohol intake and the risk of developing ER tumors. Furthermore, we observed a statistically significant interaction between alcohol intake and the use of postmenopausal hormones on the risk for ER+PR+ tumors (Pinteraction = .039). CONCLUSION: The observed association between risk of developing postmenopausal ER+ breast cancer and alcohol drinking, especially among those women who use postmenopausal hormones, may be important, because the majority of breast tumors among postmenopausal women overexpress ER. PMID- 16264181 TI - Coronary heart disease mortality after 5 years of adjuvant tamoxifen therapy: results from a randomized trial. AB - From January 1, 1983, through December 31, 1992, a total of 4610 patients entered a randomized trial that compared mortality among patients receiving 2 years of adjuvant tamoxifen therapy with that in patients receiving 5 years of adjuvant tamoxifen therapy, 4175 of whom were recurrence free after 2 years of tamoxifen therapy. Among the 2046 patients randomly assigned to the 5-year group all-cause mortality, breast cancer-specific mortality, and the incidence of contralateral breast cancer were reduced, compared with those among 2129 patients randomized in the 2-year group, but the incidence of endometrial cancer was increased. In addition, mortality from coronary heart disease was statistically significantly reduced in the 5-year group, compared with that in the 2-year group (hazard ratio = 0.67, 95% confidence interval = 0.47 to 0.94; P = .022 [two-sided Wald test]). Ten years after surgery, 2.1% of the patients in the 5-year group and 3.5% of those in the 2-year group had died from coronary heart disease. No statistically significant increases in mortality from other heart diseases, cerebrovascular diseases, or other vascular diseases were observed. PMID- 16264182 TI - Effect of gamma-linolenic acid on the transcriptional activity of the Her-2/neu (erbB-2) oncogene. AB - The omega-6 polyunsaturated fatty acid gamma-linolenic acid (GLA; 18:3n-6), which is found in several plant oils and is used as an herbal medicine, has antitumor activity in vitro. We examined the effect of GLA on the expression of the Her 2/neu (erbB-2) oncogene, which is involved in development of numerous types of human cancer. Flow cytometric and immunoblotting analyses demonstrated that GLA treatment substantially reduced Her-2/neu protein levels in the Her-2/neu overexpressing cell lines BT-474, SK-Br3, and MDA-MB-453 (breast cancer), SK-OV3 (ovarian cancer), and NCI-N87 (gastrointestinal tumor derived). GLA exposure led to a dramatic decrease in Her-2/neu promoter activity and a concomitant increase in the levels of polyomavirus enhancer activator 3 (PEA3), a transcriptional repressor of Her-2/neu, in these cell lines. In transient transfection experiments, a Her-2/neu promoter bearing a PEA3 site-mutated sequence was not subject to negative regulation by GLA in Her-2/neu-overexpressing cell lines. Concurrent treatments of Her-2/neu-overexpressing cancer cells with GLA and the anti-Her-2/neu antibody trastuzumab led to synergistic increases in apoptosis and reduced growth and colony formation. PMID- 16264183 TI - A putative exonic splicing polymorphism in the BCL6 gene and the risk of non Hodgkin lymphoma. AB - Recent studies have shown that the B-cell lymphoma 6 gene (BCL6) is an oncogene that contributes to lymphomagenesis. Exon 6 of BCL6 contains a common single nucleotide polymorphism (SNP) (-195 C>T; dbSNP ID: rs1056932) that alters a potential binding site for an exonic splicing enhancer. We used unconditional logistic regression models to examine the association between this SNP and the risk of non-Hodgkin lymphoma (NHL) in a population-based case-control study of women residing in Connecticut (461 case patients and 535 control subjects). The risk of NHL among women with the CC genotype was more than double that of women with the TT genotype (odds ratio [OR] = 2.2, 95% confidence interval [CI] = 1.5 to 3.3). Higher risks were observed for two NHL subtypes, namely B-cell chronic lymphatic leukemia/prolymphocytic leukemia/small lymphocytic lymphoma (OR = 3.5, 95% CI = 1.6 to 7.8) and T-cell lymphoma (OR = 5.2, 95% CI = 2.0 to 13.3). Our results support the hypothesis that a genetic variant that could alter mRNA transcripts of BCL6 may contribute to the etiology of NHL and suggest that this variant warrants further investigation. PMID- 16264184 TI - Re: Ethnicity and breast cancer: factors influencing differences in incidence and outcome. PMID- 16264185 TI - Re: Systematic aortic and pelvic lymphadenectomy versus resection of bulky nodes in optimally debulked advanced ovarian cancer: a randomized clinical trial. PMID- 16264186 TI - Re: Systematic aortic and pelvic lymphadenectomy versus resection of bulky nodes in optimally debulked advanced ovarian cancer: a randomized clinical trial. PMID- 16264187 TI - Eukaryotic mRNA 3' processing: a common means to different ends. PMID- 16264188 TI - Telomere identity crisis. PMID- 16264189 TI - Lost in translation: the influence of ribosomes on bacterial mRNA decay. AB - The lifetimes of bacterial mRNAs are strongly affected by their association with ribosomes. Events occurring at any stage during translation, including ribosome binding, polypeptide elongation, or translation termination, can influence the susceptibility of mRNA to ribonuclease attack. Ribosomes usually act as protective barriers that impede mRNA cleavage, but in some instances they can instead trigger the decay of the mRNA to which they are bound or send a signal that leads to widespread mRNA destabilization within a cell. The influence of translation on mRNA decay provides a quality-control mechanism for minimizing the use of poorly or improperly translated mRNAs as templates for the production of abnormal proteins that might be toxic to bacteria. PMID- 16264190 TI - The HIR corepressor complex binds to nucleosomes generating a distinct protein/DNA complex resistant to remodeling by SWI/SNF. AB - The histone regulatory (HIR) and histone promoter control (HPC) repressor proteins regulate three of the four histone gene loci during the Saccharomyces cerevisiae cell cycle. Here, we demonstrate that Hir1, Hir2, Hir3, and Hpc2 proteins form a stable HIR repressor complex. The HIR complex promotes histone deposition onto DNA in vitro and constitutes a novel nucleosome assembly complex. The HIR complex stably binds to DNA and nucleosomes. Furthermore, HIR complex binding to nucleosomes forms a distinct protein/DNA complex resistant to remodeling by SWI/SNF. Thus, the HIR complex is a novel nucleosome assembly complex which functions with SWI/SNF to regulate transcription. PMID- 16264191 TI - The Drosophila nucleosome remodeling factor NURF is required for Ecdysteroid signaling and metamorphosis. AB - Drosophila NURF is an ISWI-containing ATP-dependent chromatin remodeling complex that regulates transcription by catalyzing nucleosome sliding. To determine in vivo gene targets of NURF, we performed whole genome expression analysis on mutants lacking the NURF-specific subunit NURF301. Strikingly, a large set of ecdysone-responsive targets is included among several hundred NURF-regulated genes. Null Nurf301 mutants do not undergo larval to pupal metamorphosis, and also enhance dominant-negative mutations in ecdysone receptor. Moreover, purified NURF binds EcR in an ecdysone-dependent manner, suggesting it is a direct effector of nuclear receptor activity. The conservation of NURF in mammals has broad implications for steroid signaling. PMID- 16264192 TI - Elevated telomere-telomere recombination in WRN-deficient, telomere dysfunctional cells promotes escape from senescence and engagement of the ALT pathway. AB - Werner Syndrome (WS) is characterized by premature aging, genomic instability, and cancer. The combined impact of WRN helicase deficiency and limiting telomere reserves is central to disease pathogenesis. Here, we report that cells doubly deficient for telomerase and WRN helicase show chromosomal aberrations and elevated recombination rates between telomeres of sister chromatids. Somatic reconstitution of WRN function, but not a WRN helicase-deficient mutant, abolished telomere sister chromatid exchange (T-SCE), indicating that WRN normally represses T-SCEs. Elevated T-SCE was associated with greater immortalization potential and resultant tumors maintained telomeres via the alternative lengthening of telomere (ALT) pathway. We propose that the increased incidence of chromosomal instability and cancer in WS relates in part to aberrant recombinations between sister chromatids at telomeres, which facilitates the activation of ALT and engenders cancer-relevant chromosomal aberrations and tumor formation. PMID- 16264193 TI - The PAS/LOV protein VIVID supports a rapidly dampened daytime oscillator that facilitates entrainment of the Neurospora circadian clock. AB - A light-entrainable circadian clock controls development and physiology in Neurospora crassa. Existing simple models for resetting based on light pulses (so called nonparametric entrainment) predict that constant light should quickly send the clock to an arrhythmic state; however, such a clock would be of little use to an organism in changing photoperiods in the wild, and we confirm that true, albeit dampened, rhythmicity can be observed in extended light. This rhythmicity requires the PAS/LOV protein VIVID (VVD) that acts, in the light, to facilitate expression of an oscillator that is related to, but distinguishable from, the classic FREQUENCY/WHITE-COLLAR complex (FRQ/WCC)-based oscillator that runs in darkness. VVD prevents light resetting of the clock at dawn but, by influencing frq RNA turnover, promotes resetting at dusk, thereby allowing the clock to run through the dawn transition and take its phase cues from dusk. Consistent with this, loss of VVD yields a clock whose performance follows the simple predictions of earlier models, and overexpression of VVD restores rhythmicity in the light and sensitivity of phase to the duration of the photoperiod. PMID- 16264194 TI - Genomic analysis of LexA binding reveals the permissive nature of the Escherichia coli genome and identifies unconventional target sites. AB - Genomes of eukaryotic organisms are packaged into nucleosomes that restrict the binding of transcription factors to accessible regions. Bacteria do not contain histones, but they have nucleoid-associated proteins that have been proposed to function analogously. Here, we combine chromatin immunoprecipitation and high density oligonucleotide microarrays to define the in vivo DNA targets of the LexA transcriptional repressor in Escherichia coli. We demonstrate a near-universal relationship between the presence of a LexA sequence motif, LexA binding in vitro, and LexA binding in vivo, suggesting that a suitable recognition site for LexA is sufficient for binding in vivo. Consistent with this observation, LexA binds comparably to ectopic target sites introduced at various positions in the genome. We also identify approximately 20 novel LexA targets that lack a canonical LexA sequence motif, are not bound by LexA in vitro, and presumably require an additional factor for binding in vivo. Our results indicate that, unlike eukaryotic genomes, the E. coli genome is permissive to transcription factor binding. The permissive nature of the E. coli genome has important consequences for the nature of transcriptional regulatory proteins, biological specificity, and evolution. PMID- 16264196 TI - Studies on LXR- and FXR-mediated effects on cholesterol homeostasis in normal and cholic acid-depleted mice. AB - As previously reported by us, mice with targeted disruption of the CYP8B1 gene (CYP8B1-/-) fail to produce cholic acid (CA), upregulate their bile acid synthesis, reduce the absorption of dietary cholesterol and, after cholesterol feeding, accumulate less liver cholesterol than wild-type (CYP8B1+/+) mice. In the present study, cholesterol-enriched diet (0.5%) or administration of a synthetic liver X receptor (LXR) agonist strongly upregulated CYP7A1 expression in CYP8B1-/- mice, compared to CYP8B1+/+ mice. Cholesterol-fed CYP8B1-/- mice also showed a significant rise in HDL cholesterol and increased levels of liver ABCA1 mRNA. A combined CA (0.25%)/cholesterol (0.5%) diet enhanced absorption of intestinal cholesterol in both groups of mice, increased their liver cholesterol content, and reduced their expression of CYP7A1 mRNA. The ABCG5/G8 liver mRNA was increased in both groups of mice, but cholesterol crystals were only observed in bile from the CYP8B1+/+ mice. The results demonstrate the cholesterol-sparing effects of CA: enhanced absorption and reduced conversion into bile acids. Farnesoid X receptor (FXR)-mediated suppression of CYP7A1 in mice seems to be a predominant mechanism for regulation of bile acid synthesis under normal conditions and, as confirmed, able to override LXR-mediated mechanisms. Interaction between FXR- and LXR-mediated stimuli might also regulate expression of liver ABCG5/G8. PMID- 16264195 TI - Coronary vessel development requires activation of the TrkB neurotrophin receptor by the Wilms' tumor transcription factor Wt1. AB - The formation of intramyocardial blood vessels is critical for normal heart development and tissue repair after infarction. We report here expression of the Wilms' tumor gene-1, Wt1, in coronary vessels, which could contribute to the defective cardiac vascularization in Wt1-/- mice. Furthermore, the high-affinity neurotrophin receptor TrkB, which is expressed in the epicardium and subepicardial blood vessels, was nearly absent from Wt1-deficient hearts. Activation of Wt1 in an inducible cell line significantly enhanced TrkB expression. The promoter of NTRK2, the gene encoding TrkB, was stimulated approximately 10-fold by transient cotransfection of a Wt1 expression construct. The critical DNA-binding site for activation of the NTRK2 promoter by Wt1 was delineated by DNase I footprint analysis and electrophoretic mobility shift assay. Transgenic experiments revealed that the identified Wt1 consensus motif in the NTRK2 promoter was necessary to direct expression of a reporter gene to the epicardium and the developing vasculature of embryonic mouse hearts. Finally, mice with a disrupted Ntrk2 gene lacked a significant proportion of their intramyocardial blood vessels. These findings demonstrate that transcriptional activation of the TrkB neurotrophin receptor gene by the Wilms' tumor suppressor Wt1 is a crucial mechanism for normal vascularization of the developing heart. PMID- 16264199 TI - Release and elimination of soluble vasoactive factors during percutaneous coronary intervention of saphenous vein grafts: analysis using the PercuSurge GuardWire distal protection device. AB - BACKGROUND: The presence of no-reflow substantially increases the risk of major adverse clinical events (MACE) in percutaneous coronary artery interventions (PCI). Distal protection devices may eliminate both debris and soluble factors that can lead to no-reflow. This study was designed to evaluate the soluble factors released and eliminated by the PercuSurge GuardWire during SVG intervention. METHODS: Twenty-eight consecutive patients underwent PCI on 34 lesions in 31 SVGs using the PercuSurge GuardWire. Blood was taken prior to PCI for baseline measurement of: (1) vasoconstrictive factors: endothelin (ET) and serotonin (5-HT); (2) thrombotic factors: tissue factor (TF), plasminogen activator inhibitor (PAI-1), thrombin/antithrombin III complex (TAT), and prothrombin fragment F1+2 (F1+2); and (3) inflammatory factors: soluble CD40 ligand (sCD40L) and soluble E-selectin. After stenting and before deflating the distal protection balloon, 2 aspiration runs were performed with the PercuSurge Export Catheter and sent for analysis. RESULTS: Clinical follow-up was conducted at an average of 6 +/- 3 months. SVG PCI resulted in a substantial increase in levels of vasoconstrictive factors, including ET (3 x increase; p < 0.001) and 5 HT (9.7 x increase; p = 0.031); thrombotic factors, including TF (4.5 x increase; p = 0.005), TAT (2.4 x increase; p < 0.001), and F1+2 (2.4 x increase; p < 0.001). Pro-inflammatory factors were also released during the procedure. Levels of sCD40L and soluble E-selectin increased 123% (p = 0.003) and 25% (p < 0.001), respectively. CONCLUSIONS: SVG PCI results in an immediate increase in vasoactive factors, including vasoconstrictive, thrombotic and inflammatory factors. The PercuSurge GuardWire effectively removes these soluble factors, which may account for reduction of no-reflow and other complications during SVG interventions. PMID- 16264198 TI - Linkage analysis of LDL cholesterol in American Indian populations: the Strong Heart Family Study. AB - Previous studies have demonstrated that low density lipoprotein cholesterol (LDL C) concentration is influenced by both genes and environment. Although rare genetic variants associated with Mendelian causes of increased LDL-C are known, only one common genetic variant has been identified, the apolipoprotein E gene (APOE). In an attempt to localize quantitative trait loci (QTLs) influencing LDL C, we conducted a genome-wide linkage scan of LDL-C in participants of the Strong Heart Family Study (SHFS). Nine hundred eighty men and women, age 18 years or older, in 32 extended families at three centers (in Arizona, Oklahoma, and North and South Dakota) were phenotyped for LDL-C concentration and other risk factors. Using a variance component approach and the program SOLAR, and after accounting for the effects of covariates, we detected a QTL influencing LDL-C on chromosome 19, nearest marker D19S888 at 19q13.41 [logarithm of odds (LOD) = 4.3] in the sample from the Dakotas. This region on chromosome 19 includes many possible candidate genes, including the APOE/C1/C4/C2 gene cluster. In follow-up association analyses, no significant evidence for an association was detected with the APOE*2 and APOE*4 alleles (P = 0.76 and P = 0.53, respectively). Suggestive evidence of linkage to LDL-C was detected on chromosomes 3q, 4q, 7p, 9q, 10p, 14q, and 17q. These linkage signals overlap positive findings for lipid related traits and harbor plausible candidate genes for LDL-C. PMID- 16264197 TI - Regulation of bile acid biosynthesis by hepatocyte nuclear factor 4alpha. AB - Hepatocyte nuclear factor 4alpha (HNF4alpha) regulates many genes that are preferentially expressed in liver. Mice lacking hepatic expression of HNF4alpha (HNF4alphaDeltaL) exhibited markedly increased levels of serum bile acids (BAs) compared with HNF4alpha-floxed (HNF4alphaF/F) mice. The expression of genes involved in the hydroxylation and side chain beta-oxidation of cholesterol, including oxysterol 7alpha-hydroxylase, sterol 12alpha-hydroxylase (CYP8B1), and sterol carrier protein x, was markedly decreased in HNF4alphaDeltaL mice. Cholesterol 7alpha-hydroxylase mRNA and protein were diminished only during the dark cycle in HNF4alphaDeltaL mice, whereas expression in the light cycle was not different between HNF4alphaDeltaL and HNF4alphaF/F mice. Because CYP8B1 expression was reduced in HNF4alphaDeltaL mice, it was studied in more detail. In agreement with the mRNA levels, CYP8B1 enzyme activity was absent in HNF4alphaDeltaL mice. An HNF4alpha binding site was found in the mouse Cyp8b1 promoter that was able to direct HNF4alpha-dependent transcription. Surprisingly, cholic acid-derived BAs, produced as a result of CYP8B1 activity, were still observed in the serum and gallbladder of these mice. These studies reveal that HNF4alpha plays a central role in BA homeostasis by regulation of genes involved in BA biosynthesis, including hydroxylation and side chain beta-oxidation of cholesterol in vivo. PMID- 16264200 TI - Soluble mediators of microvascular dysfunction--identifying the invisible culprits? PMID- 16264202 TI - More stents, fewer LIMAs? PMID- 16264201 TI - Sirolimus-eluting stent treatment for complex proximal left anterior descending artery stenoses: 7-month clinical and angiographic results. AB - BACKGROUND: High-grade involvement of the proximal left anterior descending artery (LAD), either in isolation or as part of multivessel coronary artery disease, remains a frequent indication for surgical revascularization. This is particularly true in complex proximal LAD lesions among high-risk patients, since stenting is usually ineffective in the long term as regards freedom from angina and target vessel revascularization (TVR) in such patient subsets. The sirolimus eluting stent (SES) has been reported to significantly reduce clinical and angiographic restenosis rates. We therefore analyzed the clinical and angiographic long-term results of a group of complex patients treated with SES for proximal LAD stenoses. METHODS: A total of 80 consecutive patients treated with sirolimus-eluting stents (SES) for complex proximal LAD stenoses were analyzed. This high-risk cohort included diabetic patients, long, bifurcated, ostial and heavily calcified lesions, chronic total occlusions and in-stent restenoses. Late lumen loss and clinically-driven target lesion revascularization (TLR) were analyzed at 7 months. RESULTS: Late lumen loss was 0.19 +/- 0.06 mm in the analysis segment (stent with 5 mm proximal and distal to it) at 7 months. TLR was performed in 6.3% of cases, and the combined rate of death from any cause, nonfatal MI and TLR was also 6.3% at 7 months. CONCLUSION: SES, once successfully implanted into complex proximal LAD stenoses, appear effective, with rather acceptable rates of adverse events at 7-month follow-up. Accordingly, treating the proximal LAD using SES in complex patients may narrow the gap between PCI and surgery for this indication. PMID- 16264204 TI - Safety and efficacy of a multipurpose coronary angiography strategy using the transradial technique. AB - The use of a single catheter for coronary angiography has a number of potential advantages such as the reduction of arterial trauma, costs and procedural time. Accordingly, we assessed the feasibility and safety of two different strategies for transradial multipurpose coronary angiography. METHODS: From February 2002 to December 2004, a total of 657 transradial diagnostic catheterizations were performed by a single operator in which engagement of both left and right coronary arteries was attempted either with a Judkins Left 3.5 (n = 194) or a Brachial Type K (n = 463) catheter. Success rates and complications were analyzed. RESULTS: The mean age was 64.4 +/- 11.9 years. The right radial artery was used in 93% of the cases. Either left or right coronary angiography was possible in 94% of the Brachial type K cases, and in 97% of the Judkins Left cases (p = 0.11). Overall, a single-catheter procedure was possible in 87% of the cases (85% with Brachial Type K and 92% with Judkins Left 3.5; p = 0.01). There were no significant differences in fluoroscopy time (4.8 +/- 3.8 minutes versus 5.0 +/- 3.8 minutes; p = 0.61), or in dye volume (103 +/- 33 ml versus 114 +/- 78 ml; p = 0.15). There were no cases of femoral or contralateral arm crossover, and no coronary or aortic dissections or systemic embolization. CONCLUSION: A strategy of performing both left and right coronary angiography with a single Brachial Type K or Judkins Left catheter by the transradial technique is attractive and appears to be safe and effective. PMID- 16264203 TI - Myonecrosis after elective percutaneous coronary intervention: effect of clopidogrel-statin interaction. AB - BACKGROUND: A recent ex vivo study suggests that the metabolic activation of clopidogrel is catalyzed by cytochrom P450 (CYP) 3A4 and is competitively inhibited by atorvastatin, but not pravastatin. OBJECTIVE: To determine whether the incidence of procedure-related myocardial injury, assessed by cardiac troponin T (cTnT) release, is altered when clopidogrel is coadministered with a statin that is predominantly CYP3A4-metabolized. METHODS AND RESULTS: Of the 211 consecutive patients who underwent coronary stenting after pretreatment with clopidogrel, 114 were receiving a CYP3A4-metabolized statin (59 simvastatin and 55 atorvastatin, Group 1), and 37 were receiving a non-CYP3A4-metabolized statin (30 pravastatin and 7 fluvastatin, Group 2) whereas 60 patients were not taking any statins (Control). All were troponin-negative before the procedure. The overall incidence of postprocedural cTnT positivity (> 0.10 ng/ml) was 30.8%. Group 2 patients were less likely to exhibit cTnT rise relative to Group 1 patients (8% versus 41.6%; p = 0.004) and relative to controls (8% versus 32.5%; p < 0.001). Multivariate analysis identified the use of a non-CYP3A4-metabolized statin before coronary stenting as the sole independent predictor for lower incidence of procedure-related cTnT elevation with estimated Odds ratios of 0.16 relative to no statin therapy (95% CI: 0.04-0.59; p = 0.006) and 0.18 relative to CYP3A4-metabolized statin therapy (95 CI: 0.053-0.637; p = 0.008) CONCLUSION: Benefit derived from the preprocedural use of pravastatin and fluvastatin but not atorvastatin and simvastatin suggest that the ex vivo finding of a negative interaction when coadministering a CYP3A4-metabolized statin with clopidogrel may be of clinical significance. PMID- 16264205 TI - Poor outcome in patients treated with brachytherapy for diffuse in-stent restenosis. The role of additional stenting despite prolonged antiplatelet therapy. AB - BACKGROUND: Brachytherapy (IBT) has been the first effective treatment of in stent restenosis (ISR). However, when IBT is associated with additional stenting, high rates of late thrombosis have been observed. Even though prolongation of a double antiplatelet therapy seems to have overcome this problem, studies analyzing whether additional stenting still remains a negative prognostic factor for restenosis are lacking. AIM: To evaluate outcomes of patients treated for ISR with or without additional stenting and IBT followed by prolonged antiplatelet therapy. METHODS: Seventy-seven consecutive patients treated with beta radiation in 89 lesions with ISR were analyzed according to the need for deploying additional stents: 73 lesions were treated without additional stents (Group 1) and 16 lesions with one or more new stents (Group 2) because of suboptimal results or flow-limiting dissections. Double antiplatelet therapy was administered for 12 months. An angiographic follow-up was scheduled after 6 months. P-values < 0.05 were considered significant. RESULTS: Restenosis rates were 31.5% (23/73) and 62.5% (10/16) in Group 1 (G1) and Group 2 (G2), respectively (p = 0.02). The two groups did not differ for late vessel thrombosis (8 in G1 and 2 in G2). In G2, high rates of recurrence were observed in the additional stent (6/16, 37.5%; p = 0.02 versus edge restenosis and in old stent recurrence in both G1 and G2). CONCLUSIONS: The association of additional stenting with brachytherapy in treatment of ISR is characterized by poor outcomes, even if a prolonged antiplatelet therapy has been administered. These results are related to high restenosis rates observed in the additional stent. PMID- 16264206 TI - Coronary artery perforation during percutaneous coronary intervention: incidence and outcomes in the new interventional era. AB - BACKGROUND: Coronary artery perforation (CP) is a serious complication of percutaneous coronary intervention (PCI). We sought to define the incidence and outcome of CP given the advance in interventional techniques, devices and use of glycoprotein inhibitors (GP IIb/IIIa). METHODS: We retrospectively reviewed the records of patients who underwent PCI at our institution over a four-year period. The incidence of CP was derived from patient records and then confirmed by reviewing the angiogram. Perforations were classified as Type 1, 2, or 3, as previously defined. RESULTS: A total of 4,886 patients underwent PCI. Atherectomy devices were used in 329 patients and GP IIb/IIIa in 2,200 patients. Twenty-five CP were identified (0.5% incidence). Six were Type 1 (24%), 10 were Type 2 (40%), and 9 were Type 3 (36%). 13/25 (52%) of the CP were Type C Lesions, and 12/25 (48%) occurred in calcified vessels. All Type 1 perforations were caused by coronary wires and 4/6 CP occurred with the use of hydrophilic and extra stiff wires. Type 2 perforations were caused by coronary wires in 8/10 CP, and by stent deployment in 2/10. Two patients with Type 2 CP sustained a non-ST-elevation myocardial infarction. Type 3 perforations were caused by stent placement in 4/9 CP, 2/9 by atherectomy devices, and 3/9 by coronary wires. Four patients with Type 3 CP underwent pericardial drainage, 5 patients had a myocardial infarction and 2 patients died. CONCLUSION: Type 1 and 2 perforations are predominately caused by hydrophilic and stiff wires and do not require pericardial drainage or surgical intervention. Type 3 perforations are more often associated with stent and device use. A majority of Type 3 perforations can be initially managed by percutaneous methods. PMID- 16264207 TI - Coronary perforation 2006--watch for the wire. PMID- 16264208 TI - Failure of the Symbiot PTFE-covered stent to reduce distal embolization during percutaneous coronary intervention in saphenous vein grafts. AB - OBJECTIVES: To examine the effect of the Symbiot PTFE-covered stent on distal embolization during percutaneous coronary intervention (PCI) in saphenous vein grafts (SVGs). BACKGROUND: Covered stents are intended to trap friable debris and reduce distal embolization during vein graft PCI, but have consistently failed to improve clinical outcomes in randomized trials. The reasons for this lack of benefit are unclear. METHODS: Thirty patients undergoing SVG PCI, and eligible for a FilterWire distal protection device, were randomized to Symbiot or to a conventional bare-metal stent. Postprocedure, the FilterWire was removed, fixed in formalin, and photographed under high magnification. The primary end-point was embolic debris area, measured by semi-automated edge-detection analysis of digital images. Secondary end points were no-reflow, "Filter no-reflow", and distal embolization. RESULTS: There was no significant difference between the Symbiot and conventional stent groups in embolic debris area (10.5 +/- 7.2 versus 6.6 +/- 7.8 mm2; p = 0.18). No-reflow occurred in 3/14 Symbiot versus 0/16 conventional (p = 0.09), and final TIMI flow was lower in the Symbiot group (2.7 +/- 0.5 versus 3.0 +/- 0.5; p = 0.01). However, there was no difference in the incidence of Filter no-reflow (7/14 versus 7/16; p = NS) or distal embolization (9/14 versus 8/16; p = NS). Distal embolization in the Symbiot group occurred predominantly after postdilatation (7/9; p = 0.05). CONCLUSIONS: The Symbiot PTFE covered self-expanding stent does not reduce distal embolization compared to a conventional bare metal stent. Distal embolization with the Symbiot occurs almost exclusively after mandatory postdilatation. These findings may explain the failure of the Symbiot, and of covered stents in general, to improve clinical outcomes during vein graft PCI. PMID- 16264209 TI - Use of endovascular stents for the treatment of coarctation of the aorta in children and adults: immediate and midterm results. AB - OBJECTIVES: The aim of this study was to evaluate the use of endovascular stents in both native and recurrent coarctation of the aorta (CoA) in children and adults. BACKGROUND: The use of stents in CoA as an alternative to surgery or balloon angioplasty has been shown to have favorable immediate and midterm results. METHODS: Between May 1995 and February 2005, 44 patients (28 native and 16 with recoarctation after previous intervention) at a mean age of 16.9 +/- 1.8 years (range 3 months to 44 years) underwent stent implantation. Successful outcome was defined as a reduction in the peak systolic pressure gradient by 50% or more. RESULTS: Stents were implanted in all 44 patients and successful outcomes occurred in all 44 patients. The peak systolic gradient decreased from a mean value of 29.2 +/- 1.9 mmHg (range from 7 to 55 mmHg) to a mean of 3.7 +/- 0.7 mmHg (range from 0 to 18 mmHg). Coarctation site diameter increased from a mean of 6.1 +/- 0.5 mm to a mean of 13 +/- 0.5 mm. Twelve patients underwent stent redilatation/repeat stent placement procedures between 7 to 47.5 months after the initial procedure. One patient underwent a repeat procedure after 1 day secondary to stent migration from the first procedure. Complications occurred in 9 patients, of which no patients required surgery. At a mean follow-up of 19.8 +/ 3.5 months (range 0.1 to 117 months), 1 patient developed an aneurysm at the site of stent implantation. This patient is awaiting surgical repair of the aneurysm. No other complications were noted. Therefore, we conclude that stent implantation for coarctation of the aorta is safe and effective with sustained good midterm results. PMID- 16264210 TI - Single coronary artery with the absence of a left anterior descending artery. AB - Anomalous origin of coronary arteries is discovered incidentally during coronary arteriography or at autopsy, and awareness among angiographers is required. We describe a case with a rare combination of a single coronary artery originating from the right sinus of Valsalva associated with an absent left anterior descending artery and a secundum-type atrial septal defect. PMID- 16264212 TI - Inadvertent detachment of an entrapped Cutting Balloon from the balloon catheter during treatment of in-stent restenosis. AB - We report on a case of anomalous right coronary artery with interarterial course that caused severe symptoms and was treated percutaneously after failed bypass surgery. Using intravascular ultrasound guidance, a drug-eluting stent was placed at the ostium of the anomalous artery, correcting both the baseline narrowing and the phasic, systolic arterial compression. PMID- 16264211 TI - Feasibility assessment of aortic valve area in patients with aortic stenosis using a pressure wire through a 4 French system and single femoral arterial access. AB - OBJECTIVE: We sought to determine the safety, success and accuracy of using a pressure wire through a 4 French catheter with single arterial access to determine the aortic valve area in patients with aortic stenosis. BACKGROUND: Conventional invasive techniques to assess aortic stenosis are associated with procedural risks secondary to bilateral femoral access or the sheath size and reduced accuracy using femoral sheath pressure to replace ascending aortic pressure. METHODS: Cardiac output was measured using a pulmonary artery catheter. With a 4 French catheter through single femoral access ascending aortic pressure and, with a pressure wire, left ventricular pressure were recorded simultaneously in four patients with aortic stenosis. Likewise, both pressures were recorded using the pullback method. Pressure gradients were determined by both methods and aortic valve area calculated with the Gorlin equation prior to correlation. Of note, patients with significant arrhythmias such as atrial fibrillation or frequent premature atrial or ventricular contractions were excluded to maintain accuracy of the pullback method. RESULTS: All hemodynamic parameters were successfully obtained in all four patients. The correlation between pressure wire and pullback method was highly significant in this case series (n = 4, r = 0.983, p = 0.017). There were no complications. CONCLUSION: The results suggest that the aortic valve area can be safely and accurately evaluated with a pressure wire using a 4 French system. This novel method could be the preferred method for patients with vascular access limitations. PMID- 16264213 TI - "Buddy wire" technique to overcome proximal coronary tortuosity during rotational atherectomy. AB - We report a case in which rotational atherectomy was planned for the treatment of a severely calcified obstructive lesion in the middle right coronary artery. Severe proximal vessel tortuosity prevented the advancement of the Rotablator burr. We utilized the "buddy wire" technique, allowing facilitated advancement of the Rotablator and successful atherectomy and stenting. We propose this old technique as an alternative method to allow advancement of the Rotablator burr through tortuous and calcified vessels. PMID- 16264214 TI - Intravascular ultrasound to guide stenting of an anomalous right coronary artery coursing between the aorta and pulmonary artery. AB - We report on a case of anomalous right coronary artery with interarterial course which caused severe symptoms and was treated percutaneously after failed bypass surgery. Using intravascular ultrasound (IVUS) guidance, a drug eluting stent was placed at the ostium of the anomalous artery, correcting both the baseline narrowing and the phasic, systolic arterial compression. PMID- 16264215 TI - Rescue percutaneous coronary intervention: a review. PMID- 16264216 TI - The "zipper" lesion: a rare but serious guiding catheter-induced complication of a PCI procedure. PMID- 16264217 TI - Transcatheter fenestration of autologous pericardial extracardiac Fontan via the transhepatic approach. AB - We describe a case of a patient with hypoplastic left heart syndrome (HLHS) who developed protein-losing enteropathy (PLE) following autologous pericardial extracardiac Fontan (APEF) operation with successful resolution of PLE following transcatheter fenestration via the transhepatic approach. PMID- 16264218 TI - Acute myocardial infarction during puerperium. Report of two cases of multivessel involvement treated with primary coronary intervention. PMID- 16264219 TI - Global analysis of gene expression by differential display: a mathematical model. AB - Differential display (DD) is one of the most commonly used approaches for identifying differentially expressed genes. However, there has been lack of an accurate guidance on how many DD polymerase chain reaction (PCR) primer combinations are needed to display most of the genes expressed in a eukaryotic cell. This study critically evaluated the gene coverage by DD as a function of the number of arbitrary primers, the number of 3' bases of an arbitrary primer required to completely match an mRNA target sequence, the additional 5' base match(s) of arbitrary primers in first-strand cDNA recognition, and the length of mRNA tails being analyzed. The resulting new DD mathematical model predicts that 80-160 arbitrary 13mers, when used in combinations with three one-base anchored oligo-dT primers, would allow any given mRNA within a eukaryotic cell to be detected with a 74-93% probability, respectively. The prediction was supported by both computer simulation of the DD process and experimental data from a comprehensive fluorescent DD screening for target genes of tumor-suppressor p53. Thus, this work provides a theoretical foundation upon which global analysis of gene expression by DD can be pursued. PMID- 16264221 TI - Ordered differential display. AB - Ordered differential display (ODD) is one of the approaches that uses systematic, rather than random, sampling of transcripts for display and thereby provides means to browse through essentially all the transcripts in the compared mRNA pools. It is specifically adapted for small amounts of starting material. The protocol outlined here, in addition to ODD procedure itself, also describes isolation of RNA and synthesis of double-stranded cDNA from small biological samples. PMID- 16264220 TI - Automation of fluorescent differential display with digital readout. AB - Since its invention in 1992, differential display (DD) has become the most commonly used technique for identifying differentially expressed genes because of its many advantages over competing technologies such as DNA microarray, serial analysis of gene expression (SAGE), and subtractive hybridization. Despite the great impact of the method on biomedical research, there has been a lack of automation of DD technology to increase its throughput and accuracy for systematic gene expression analysis. Most of previous DD work has taken a "shot gun" approach of identifying one gene at a time, with a limited number of polymerase chain reaction (PCR) reactions set up manually, giving DD a low-tech and low-throughput image. We have optimized the DD process with a new platform that incorporates fluorescent digital readout, automated liquid handling, and large-format gels capable of running entire 96-well plates. The resulting streamlined fluorescent DD (FDD) technology offers an unprecedented accuracy, sensitivity, and throughput in comprehensive and quantitative analysis of gene expression. These major improvements will allow researchers to find differentially expressed genes of interest, both known and novel, quickly and easily. PMID- 16264222 TI - GeneCalling: transcript profiling coupled to a gene database query. AB - We describe here the GeneCalling method for the discovery of differentially expressed genes, both known and novel, from any species and with useful sequence information to determine the potential function of novel genes captured. The method relies on transcript visualization coupled to a database query to rapidly and quantitatively identify differentially expressed transcripts. The method has been applied to a wide variety of disease models in a wide variety of species, addressing problems as diverse as identifying novel human cancer gene targets, understanding how drugs and diet affect animal models of disease, and understanding the basis of trait differences in related strains of corn. PMID- 16264223 TI - High-density sampling differential display of prokaryotic mRNAs with RAP-PCR. AB - A high-throughput approach to prokaryotic differential display has been developed. A large number of reverse transcription polymerase chain reactions (RT PCR) are performed on total RNA isolated from induced and control bacterial cultures. Each RT-PCR reaction uses a single oligonucleotide primer and constitutes an independent sampling of the mRNA population. The large number of reactions performed allows the repeated sampling of the targeted polycistronic mRNA, which is clearly identified among possible false positives. PMID- 16264224 TI - Vertical arrays: microarrays of complex mixtures of nucleic acids. AB - Vertical arrays are microarrays that have complex mixtures of nucleic acids as array elements, and that are hybridized with single sequence probes. Like dot blots, many different experiments can be spotted on a single vertical array, allowing single genes to be compared across many conditions. Vertical arrays have two additional advantages over dot blots. First, they are printed on glass slides, allowing the use of low-volume, high-concentration hybridization reactions. Second, they can be made using low-complexity representations of the original nucleic acid population. This increases signal-to-noise relative to the usual use of dot blots, wherein the entire complexity of the population is usually spotted. Whereas standard microarrays achieve horizontal coverage of many genes and are repeated to cover many experiments, vertical arrays achieve vertical coverage of many experiments and are repeated to cover many genes. In cases where the number of genes is limited, but the number of experiments is very large, vertical arrays may be advantageous. PMID- 16264225 TI - Automated pattern ranking in differential display data analysis. AB - Gene expression analysis by differential display (DD) is limited by the labor intensive visual evaluation of the electrophoretic data traces. We describe a flexible method for computer-assisted ranking of expression patterns in data from DD experiments. The method is based on a pairwise alignment and comparison of the quantitative trace data with respect to specific expression patterns defined by the investigator. The observed patterns are ranked according to a score value that identifies the most potential findings to be confirmed visually instead of the vast amount of original results. This two-step approach, enabled by the efficient computer algorithm for gene expression pattern comparison, will increase the percentage of true-positive findings chosen for the tedious downstream processing, while minimizing the cost and labor involved in large scale DD data analysis. PMID- 16264227 TI - Differentially expressed genes associated with hepatitis B virus HBx and MHBs protein function in hepatocellular carcinoma. AB - HBx and MHBst products from hepatitis B virus-DNA (HBV-DNA), which become transcriptional transactivators of cellular and viral genes, are known to play causative roles in the development of hepatocellular carcinoma (HCC). However, the biomolecular mechanism(s) for their roles in hepatocarcinogenesis in vivo remain poorly understood. To identify authentic cellular genes involved in HBx and MHBst-transactivated carcinogenesis,we used mRNA differential display polymerase chain reaction (DD-PCR). We examined HBx and MHBs-positive or negative HCC, which had chromosomally integrated HBV DNA, vs nontumor tissues, respectively, and differentially expressed genes in either type of HCC were identified and compared with each other. Using 240 different combinations of three one-base anchored oligo-dT primers and 80 arbitrary 13mers, 16 genes were differentially expressed in the HBx and MHBs-positive HCC including RoRNA hY1, glutamine synthetase, factor H homologue 3' end, voltage-dependent anionc hannel 3 (VDAC3), three ribosomal proteins, four mitochondrial genes, and four novel genes. Unexpectedly, upregulated genes in association with functional HBV proteins were different from those reportedly transactivated by HBV viral proteins in vitro. Ten genes were downregulated, including three novel genes. In contrast, 15 genes in HCC tissue negative for HBx and MHBs-expression were preferentially expressed including pancreatic secretory trypsin inhibitor (PSTI), H19, guanidine nucleotide-binding protein alpha-1 subunit (GNAZ), carbamyl phosphate synthetase I (CPS I), insulin-like growth factor (IGF)-II, and 10 ribosomal proteins genes. Eighteen genes were downregulated including acute phase genes, a novel gene, and particularly the retinoblastoma susceptibility gene. Only two genes (ribosomal protein P0 and L37a) were commonly upregulated in both types of HCC tissues. These results suggest that cellular genes involved in the viral protein-transactivation may generally differ from those not associated with transactivation in established HCC, and that the specific oncogenic coordination through the transactivation by viral proteins which works in experiments in vitro, may play only a potential role in hepatocarcinogenesis in vivo. In addition, the functional analyses of the eight novel genes identified in this study might be valuable to further understand the mechanism(s) of hepatocarcinogenesis. PMID- 16264226 TI - Linking cDNA-AFLP-based gene expression patterns and ESTs. AB - Massive amounts of DNA sequence data, generated from expressed sequence tag (EST) and genome sequencing projects, require efficient methods to link sequence databases with temporal and spatial expression profiles. To meet this need, we have developed a powerful computer program (GenEST), which links cDNA sequence data (including EST sequences) with transcript profiles revealed by cDNA amplified fragment length polymorphism (AFLP). cDNA-AFLP is a highly reproducible differential display method based on restriction enzyme digests and selective amplification under high stringency conditions. GenEST predicts the sizes of virtual transcript derived fragments (TDFs) from cDNA sequences digested in silico. The resulting virtual TDFs could be traced back among the thousands of TDFs displayed on cDNA-AFLP gels. As a consequence, cDNA sequence databases can be screened very efficiently to identify genes with relevant expression profiles. Vice versa, using the restriction enzyme recognition sites, the primer extensions and the estimated TDF size as identifiers, the DNA sequence(s) corresponding to a TDF with an interesting expression pattern can be identified. PMID- 16264228 TI - Identification of disease markers by differential display: prion disease. AB - In order to identify molecular markers of prion disease in peripheral tissues, we used the differential display reverse-transcriptase polymerase chain reaction (DDRT-PCR) procedure to compare gene expression in spleens of infected and uninfected mice. In this study, we identified a novel erythroid-specific gene that was differentially expressed as a result of prion infection. We were able to demonstrate that a decrease in the expression levels of this transcript in hematopoietic tissues was a common feature of prion diseases. Our findings suggest a previously unknown role for the blood erythroid lineage in the development of prion diseases and should provide a new focus for research into diagnostic and therapeutic strategies. PMID- 16264229 TI - Saturation screening for p53 target genes by digital fluorescent differential display. AB - Differential display (DD) is one of the most commonly used approaches for identifying differentially expressed genes. Despite the great impact of the method on biomedical research, there has been a lack of automation of DD technology to increase its throughput and accuracy for a systematic gene expression analysis. Most of previous DD work has taken a "shotgun" approach of identifying one gene at a time, with limited polymerase chain reaction (PCR) reactions set up manually, giving DD a low-technology and low-throughput image. With our newly created DD mathematical model, which has been validated by computer simulations, global analysis of gene expression by DD technology is no longer a shot in the dark. After identifying the "rate-limiting" factors that contribute to the "noise" level of DD method, we have optimized the DD process with a new platform that incorporates fluorescent digital readout and automated liquid handling. The resulting streamlined fluorescent DD (FDD) technology offers an unprecedented accuracy, sensitivity, and throughput in comprehensive and quantitative analysis of gene expression. We are using this newly integrated FDD technology to conduct a systematic and comprehensive screening for p53 tumor suppressor gene targets. PMID- 16264230 TI - Identification of p53-regulated genes by the method of differential display. AB - The p53 tumor-suppressor gene is mutated in a wide range of human cancers. The ability of p53 to control passage through the cell cycle (in G1 and in G2) and to control apoptosis in response to abnormal proliferative signals and stress, including DNA damage, is considered to be important for its tumor-suppression function. p53 is a transcription factor that binds to DNA in a sequence-specific manner to activate transcription of target genes. In this chapter, we describe the application of differential display to identify p53-regulated genes. PMID- 16264231 TI - Identification by differential display of IL-24 autocrine loop activated by ras oncogenes. AB - Ras signaling pathway is thought to control the expression of a subset of yet-to be-defined genes that are crucial for cell growth and differentiation. Here we have identified by differential display a novel oncogenic Ras target gene encoding a new cytokine. Biochemical studies reveal that this cytokine, which we named IL-24, is a member of IL-10 family of cytokines, and it signals through two hetorodimeric receptors, whose expression is also upregulated by ras oncogenes. Thus, IL-24 and its receptors may represent a novel autocrine loop coordinately activated by ras oncogenes. PMID- 16264233 TI - Functional analysis of nocturnin: a circadian clock-regulated gene identified by differential display. AB - Within the retina there is a circadian clock that controls the 24-h timing of processes such as hormone release, cell movement, and gene transcription. In an effort to better understand the molecular nature of this retinal clock, a differential display (DD) screen was performed to isolate a gene with high amplitude circadian rhythmicity in the Xenopus retina. A novel gene expressed in the early evening in photoreceptor cells was isolated and named nocturnin for night factor. This article outlines the steps we took to study a protein of unknown function, particularly highlighting the analyses one can perform when little more than the primary sequence of a gene is known. In addition, we describe the results of sequence analysis that assisted in predicting the function of nocturnin. We have shown that nocturnin acts as a deadenylase in vitro, removing the poly(A) tail from a mature messenger RNA in a process that either leads to degradation or translational silencing of a message. Although the role of nocturnin in the retina is unknown, future studies to identify target mRNAs that are deadenylated by nocturnin will assist in elucidating its physiological role in this tissue. PMID- 16264232 TI - Comprehensive analysis of ovarian gene expression during ovulation using differential display. AB - Mammalian ovulation is a normal biological process that is initiated when a gonadotropic hormone stimulates G protein-coupled receptors in the plasma membrane of cells in ovarian follicles. This article outlines differential display (DD) protocols and associated methods that have been used to discover more than 30 genes that are expressed in the rat ovary during the ovulatory process. Details are provided regarding the methods for total RNA extraction, reverse transcription (RT), DD-polymerase chain reaction (PCR), polyacrylamide gel electrophoresis (PAGE), Northern analysis of the differentially expressed cDNA fragments, cloning of the cDNA fragments, sequencing of the cDNA, and in situ hybridization of the cDNA fragments with sections of ovarian tissue. These methods provide clear evidence of the temporal and spatial patterns of expression of ovulation-specific genes in the ovary. Most of the genes that have been discovered to date have been associated previously with cascades of gene expression in acute inflammatory reactions. Therefore, the data support the working hypothesis that the ovary becomes inflamed at the time of ovulation, and this acute condition softens local connective tissues and causes ovarian follicles to rupture and release fertile eggs. PMID- 16264234 TI - Isolation and characterization of anthocyanin 5-O-glucosyltransferase in Perilla frutescens var. crispa by differential display. AB - The glucosylation of 5-position of anthocyanin molecules is one of most important modification step, which is responsible for the stability and color hue of anthocyanin molecules. However, anthocyanin 5-O-glucosyltransferase (5-GT) is unstable and it had been difficult to purify this enzyme by biochemical methods. The isolation of cDNA encoding 5-GT was succeeded first by mRNA differential display (DD) of red and green chemotypes of Perilla frutescens var. crispa. The full-length cDNA was isolated by the screening of perilla cDNA library with cDNA fragments, which was displayed specifically in red perilla. The biological characters of this enzyme were investigated by using the recombinant protein expressed in yeast. We have shown an example of the application of DD technique to the molecular biology in plant secondary metabolisms. PMID- 16264235 TI - Identification of target genes of a yeast transcriptional repressor. AB - Xbp1 is a transcriptional repressor with homology to the DNA binding domains of two cell-cycle regulatory transcription factors, Swi4 and Mbp1. Target genes bound and regulated by Xbp1, have been identified using differential display (DD). Because little was known about the regulation or function of Xbp1, we identified genes that were underrepresented in RNAs purified from cells in which Xbp1 was ectopically expressed. Four target genes, including three cyclin genes, were identified by DD and then confirmed by binding studies and Northern analysis. The binding site for Xbp1, previously identified by site selection, was present in these target genes and is conserved phylogenetically. PMID- 16264236 TI - Detection of an mRNA polymorphism by differential display. AB - Differential display technology was utilized to compare programs of gene expression in primary cultures of human skin fibroblasts from normal volunteers and patients diagnosed with melancholic depression. Polymorphic transcripts of a single gene differing by one tandem repeat sequence of four nucleotides (TGAT) in the 3' noncoding region were detected. PMID- 16264237 TI - Silencing in yeast: identification of clr4 targets. AB - Efficient handling of multiple reactions is a crucial prerequisite for productive RNA differential display (DD) analysis. To identify transcriptional targets of the histone H3 Lys9-specific methyltransferase Clr4, we applied a multiformat modification of DD to compare between clr4+ and clr4- transcriptomes of Schizosaccaromyces pombe. As a result, 14 differentially expressed bands were identified among 720 polymerase chain reaction (PCR) studied. The content of these bands was then analyzed by cloning, sequencing, and Northern analysis. In the final stage of verification, four Clr4 targets were isolated based on their expression in six Clr4 chromo and SET domain mutant strains. The step-by-step description of the multiformat DD provided below includes RNA purification, cDNA synthesis, 96-well PCR, electrophoretic separation of PCR products, isolation of DNA fragments from differentially expressed bands, and verification of candidate genes by Northern analysis. PMID- 16264238 TI - Identification of mRNA bound to RNA binding proteins by differential display. AB - A large number of RNA binding proteins have recently been identified that influence various human genetic disorders. However, the specific function of many of these proteins and what role they may play in a particular disease remains unclear. Identification of the substrate mRNA bound by an RNA binding protein will provide insights into the function of that protein and how its aberrant expression could lead to a disease phenotype. We have developed a technique termed SNAAP, for isolation of specific nucleic acids associated with proteins, to identify natural mRNA substrates for an RNA binding protein. The technique couples affinity purification of specific mRNAs bound by an RNA binding protein, with the identification of that mRNA using differential display (DD). Methods are described herein for the isolation and identification of endogenous mRNAs bound by any RNA binding protein, as well as methodology to validate the specificity of the binding. The availability of technologies to isolate the cognate substrate mRNAs potentially bound and regulated by an RNA binding protein involved in genetic disorders will greatly expedite our etiological understanding of the disorder and provide modalities for intervention. PMID- 16264239 TI - [Sexual abuse--improving awareness and responses through better education]. PMID- 16264241 TI - [The efficacy of high frequency ventilation in severe neonatal respiratory failure]. AB - OBJECTIVE: To evaluate the efficacy of high frequency ventilation (HFV) in infants failing conventional ventilator therapy at our institution. STUDY GROUP AND METHODS: Medical records of all infants managed on HFV after having failed conventional ventilator management from 1994-2004 were reviewed. Ventilatory settings, blood gases and pH just prior to starting HFV, and two and four hours after starting HFV were recorded. RESULTS: Sixty one infants met the study criteria. At two hours of HFV there was a significant improvement in oxygenation (Alveolar to arterial oxygen tension difference), ventilation and acid-base balance. These values were not significantly different between two and four hours of HFV. There was no significant difference in oxygenation between survivors (n=41) and non-survivors (n=20) prior to HFV, but after two hours of HFV the survivors had significant improvement in oxygenation. Thirty one of the survivors had improved oxygenation at two and four hours of HFV, but only eight of the nonsurvivors (p=0.03). CONCLUSIONS: HFV results in significant improvements in oxygenation, ventilation and acid-base balance in most infants failing conventional ventilatory management. The immediate response to HFV may be a predictor of survival in infants with severe hypoxic respiratory failure. PMID- 16264242 TI - [The eye of technology and the well being of women and men in Icelandic work places]. AB - INTRODUCTION: The study assessed the association between working under surveillance and electronic performance monitoring and the well-being among women and men in six Icelandic workplaces. METHODS: In the time period from February to April 2003, a questionnaire based on the General Nordic Questionnaire for Psychological and Social Factors at Work was delivered to 1369 employees in six companies where different methods of electronic performance monitoring (EPM) are used. The data was analyzed using odds ratio and logistical regression. RESULTS: The response rate was 72%, with close to equal participation of men and women. The employees who were working under EPM were more likely to have poor psychosocial work-environment, to have experienced significant stress recently, to be mentally exhausted at the end of the workday, to have significant sleep difficulties and to be dissatisfied in their job. CONCLUSION: The development of the information and communication technology that allows employers and managers to monitor and collect different electronic data about the work process and productivity of the workers makes it important to follow the health condition of those who work under electronic performance monitoring. PMID- 16264243 TI - [Prevalence of myotonic dystrophy in Iceland]. AB - OBJECTIVE: Epidemiologic studies of Myotonic Dystrophy (Dystrophic Myotony, DM) have shown variable regional prevalence from 0,46 to 189/105. We carried out a total population survey of DM in Iceland in 2004 having Oct. 31 as the day of prevalence. MATERIAL AND METHODS: Patients were collected from multiple sources, including Landspitali University Hospital registry and through contact with neurologists, neuropaediatricians, paediatricians and rehabilitation specialists. All EMGs of DM patients were reviewed. Information was gathered about age, age of onset, family history of DM and clinical symptoms. RESULTS: Eighty-two patients were ascertained giving a crude prevalence of 28.2/105. The prevalence of the congenital form of DM was 7.9/105 (23 patients, 26%). Affected females outnumbered males with a gender ratio of 1.2:1 (NS). Mean age of onset of symptoms for those, who didn't have the congenital form was 27.5 years (range 5 70 years). Ten families with DM were identified and all prevalent patients belonged to those families. CONCLUSION: The prevalence of DM is high in Iceland and higher than generally reported. This study showed a three times higher total prevalence and a seven times higher prevalence of congenital DM than found in a previous study in Iceland. We believe that this increase in prevalence probably reflects increased awareness of inherited diseases in neonates and better detection of patients who have mild symptoms. PMID- 16264244 TI - [Uniparental disomy of chromosome 11 in a patient with Beckwith-Wiedemann syndrome. First reported case in Iceland]. AB - Beckwith-Wiedemann syndrome (BWS) is a generalized overgrowth condition as well as regional and organ overgrowth in newborn children. It includes an increased risk of certain embryonal tumours. The aetiology of BWS is complex as different genetic and epigenetic alterations at chromosome region 11p15.5 may occur. We report the first case of paternal uniparental disomy in Beckwith-Wiedemann syndrome in Iceland. The diagnosis of Beckwith-Wiedemann syndrome is important as the risk of malignant tumors makes it mandatory that the children are followed for several years with regular investigations to detect the tumors as early as possible. PMID- 16264245 TI - Automated segmentation of pigmented skin lesions in multispectral imaging. AB - The aim of this study was to develop an algorithm for the automatic segmentation of multispectral images of pigmented skin lesions. The study involved 1700 patients with 1856 cutaneous pigmented lesions, which were analysed in vivo by a novel spectrophotometric system, before excision. The system is able to acquire a set of 15 different multispectral images at equally spaced wavelengths between 483 and 951 nm. An original segmentation algorithm was developed and applied to the whole set of lesions and was able to automatically contour them all. The obtained lesion boundaries were shown to two expert clinicians, who, independently, rejected 54 of them. The 97.1% contour accuracy indicates that the developed algorithm could be a helpful and effective instrument for the automatic segmentation of skin pigmented lesions. PMID- 16264246 TI - Prototype heel effect compensation filter for cone-beam CT. AB - The prototype cone-beam CT (CBCT) has a larger beam width than the conventional multi-detector row CT (MDCT). This causes a non-uniform angular distribution of the x-ray beam intensity known as the heel effect. Scan conditions for CBCT tube current are adjusted on the anode side to obtain an acceptable clinical image quality. However, as the dose is greater on the cathode side than on the anode side, the signal-to-noise ratio on the cathode side is excessively high, resulting in an unnecessary dose amount. To compensate for the heel effect, we developed a heel effect compensation (HEC) filter. The HEC filter rendered the dose distribution uniform and reduced the dose by an average of 25% for free air and by 20% for CTDI phantoms compared to doses with the conventional filter. In addition, its effect in rendering the effective energy uniform resulted in an improvement in image quality. This new HEC filter may be useful in cone-beam CT studies. PMID- 16264247 TI - Inversion recovery measurements in the presence of radiation damping and implications for evaluating contrast agents in magnetic resonance. AB - Relaxation measurements performed at high magnetic field in magnetic resonance (MR) may be adversely affected by the influence of radiation damping in concentrated samples such as water. We consider how the measured value of T1 is affected by this phenomenon for a gadolinium-doped water sample and for an undoped water sample and consider the implications for evaluating contrast agents. A simple method involving the application of a pulsed field gradient to de-phase residual transverse components of the magnetization is shown to be an effective method for suppressing this effect. Given the central role that measurement of the T1 of water plays in the assessment of contrast agents as well as a host of other MR applications, care should always be employed when measuring and interpreting T1 measurements at high magnetic fields. PMID- 16264248 TI - Small animal SPECT and its place in the matrix of molecular imaging technologies. AB - Molecular imaging refers to the use of non-invasive imaging techniques to detect signals that originate from molecules, often in the form of an injected tracer, and observe their interaction with a specific cellular target in vivo. Differences in the underlying physical principles of these measurement techniques determine the sensitivity, specificity and length of possible observation of the signal, characteristics that have to be traded off according to the biological question under study. Here, we describe the specific characteristics of single photon emission computed tomography (SPECT) relative to other molecular imaging technologies. SPECT is based on the tracer principle and external radiation detection. It is capable of measuring the biodistribution of minute (<10(-10) molar) concentrations of radio-labelled biomolecules in vivo with sub-millimetre resolution and quantifying the molecular kinetic processes in which they participate. Like some other imaging techniques, SPECT was originally developed for human use and was subsequently adapted for imaging small laboratory animals at high spatial resolution for basic and translational research. Its unique capabilities include (i) the ability to image endogenous ligands such as peptides and antibodies due to the relative ease of labelling these molecules with technetium or iodine, (ii) the ability to measure relatively slow kinetic processes (compared with positron emission tomography, for example) due to the long half-life of the commonly used isotopes and (iii) the ability to probe two or more molecular pathways simultaneously by detecting isotopes with different emission energies. In this paper, we review the technology developments and design tradeoffs that led to the current state-of-the-art in SPECT small animal scanning and describe the position SPECT occupies within the matrix of molecular imaging technologies. PMID- 16264249 TI - Comparative analyses of linac and Gamma Knife radiosurgery for trigeminal neuralgia treatments. AB - Dedicated linac-based radiosurgery has been reported for trigeminal neuralgia treatments. In this study, we investigated the dose fall-off characteristics and setup error tolerance of linac-based radiosurgery as compared with standard Gamma Knife radiosurgery. In order to minimize the errors from different treatment planning calculations, consistent imaging registration, dose calculation and dose volume analysis methods were developed and implemented for both Gamma Knife and linac-based treatments. Intra-arc setup errors were incorporated into the treatment planning process of linac-based deliveries. The effects of intra-arc setup errors with increasing number of arcs were studied and benchmarked against Gamma Knife deliveries with and without plugging patterns. Our studies found equivalent dose fall-off properties between Gamma Knife and linac-based radiosurgery given a sufficient number of arcs (>7) and small intra-arc errors (<0.5 mm) were satisfied for linac-based deliveries. Increasing the number of arcs significantly decreased the variations in the dose fall-off curve at the low isodose region (e.g. from 40% to 10%) and also improved dose uniformity at the high isodose region (e.g. from 70% to 90%). As the number of arcs increased, the effects of intra-arc setup errors on the dose fall-off curves decreased. Increasing the number of arcs also reduced the integral dose to the distal normal brain tissues. In conclusion, linac-based radiosurgery produces equivalent dose fall-off characteristics to Gamma Knife radiosurgery with a high number of arcs. However, one must note the increased treatment time for a large number of arcs and isocentre accuracies. PMID- 16264250 TI - Monte Carlo simulations of a nozzle for the treatment of ocular tumours with high energy proton beams. AB - By the end of 2002, 33 398 patients worldwide had been treated with proton radiotherapy, 10 829 for eye diseases. The dose prediction algorithms used today for ocular proton therapy treatment planning rely on parameterizations of measured proton dose distributions, i.e., broad-beam and pencil-beam techniques, whose predictive capabilities are inherently limited by severe approximations and simplifications in modelling the radiation transport physics. In contrast, the Monte Carlo radiation transport technique can, in principle, provide accurate predictions of the proton treatment beams by taking into account all the physical processes involved, including coulombic energy loss, energy straggling, multiple Coulomb scattering, elastic and nonelastic nuclear interactions, and the transport of secondary particles. It has not been shown, however, whether it is possible to commission a proton treatment planning system by using data exclusively from Monte Carlo simulations of the treatment apparatus and a phantom. In this work, we made benchmark comparisons between Monte Carlo predictions and measurements of an ocular proton treatment beamline. The maximum differences between absorbed dose profiles from simulations and measurements were 6% and 0.6 mm, while typical differences were less than 2% and 0.2 mm. The computation time for the entire virtual commissioning process is less than one day. The study revealed that, after a significant development effort, a Monte Carlo model of a proton therapy apparatus is sufficiently accurate and fast for commissioning a treatment planning system. PMID- 16264252 TI - Respiratory correlated cone-beam computed tomography on an isocentric C-arm. AB - A methodology for 3D image reconstruction from retrospectively gated cone-beam CT projection data has been developed. A mobile x-ray cone-beam device consisting of an isocentric C-arm equipped with a flat panel detector was used to image a moving phantom. Frames for reconstruction were retrospectively selected from complete datasets based on the known rotation of the C-arm and a signal from a respiratory monitor. Different sizes of gating windows were tested. A numerical criterion for blur on the reconstructed image was suggested. The criterion is based on minimization of an Ising energy function, similar to approaches used in image segmentation or restoration. It is shown that this criterion can be used for the determination of the optimal gating window size. Images reconstructed from the retrospectively gated projection sequences using the optimal gating window data showed a significant improvement compared to images reconstructed from the complete projection datasets. PMID- 16264251 TI - Fricke gel as a tool for dose distribution verification: optimization and characterization. AB - With the introduction of conformal techniques in radiation therapy, gel dosimetry plays an important role as a 3D dose verification system. There are two main types of gels in use for dosimetry: Fricke gels and polymer gels. The advantages of polymer gels are improved dose response and stability with no diffusion problems. However, the more complicated fabrication procedure and the greater cost compared to Fricke gels makes polymer gels less attractive in routine clinical use. Dose resolution has recently been introduced as a concept for comparing and optimizing the performance of different types of gel dosimeters. This parameter has not yet been investigated for Fricke gels. In this study, the effect on the dose resolution and the diffusion from different gelatine- and Fe2+ concentrations and different pH was evaluated. Increasing the concentration of gelatine from 6 wt% to 10 wt% influenced the diffusion coefficient the most, while reducing the pH from 2.0 to 1.5 had the largest effect on the dose resolution. For a gel consisting of 10 wt% gelatine, 1.0 mM Fe2+ and pH 1.5 the diffusion coefficient was found to be 1.5 mm2 h-1 and the dose resolution was about 4.1% (at 95% confidence level), for a dose of 40 Gy. By evaluating different dose gradients by the gamma-method, the diffusion was shown to have no clinically relevant impact on the dose distribution and plan acceptance within 3 h of irradiation. The results indicate a potential use of Fricke gels for IMRT verification. PMID- 16264253 TI - A theoretical comparison of two optimization methods for radiofrequency drive schemes in high frequency MRI resonators. AB - In this paper, numerical simulations are used in an attempt to find optimal source profiles for high frequency radiofrequency (RF) volume coils. Biologically loaded, shielded/unshielded circular and elliptical birdcage coils operating at 170 MHz, 300 MHz and 470 MHz are modelled using the FDTD method for both 2D and 3D cases. Taking advantage of the fact that some aspects of the electromagnetic system are linear, two approaches have been proposed for the determination of the drives for individual elements in the RF resonator. The first method is an iterative optimization technique with a kernel for the evaluation of RF fields inside an imaging plane of a human head model using pre-characterized sensitivity profiles of the individual rungs of a resonator; the second method is a regularization-based technique. In the second approach, a sensitivity matrix is explicitly constructed and a regularization procedure is employed to solve the ill-posed problem. Test simulations show that both methods can improve the B(1) field homogeneity in both focused and non-focused scenarios. While the regularization-based method is more efficient, the first optimization method is more flexible as it can take into account other issues such as controlling SAR or reshaping the resonator structures. It is hoped that these schemes and their extensions will be useful for the determination of multi-element RF drives in a variety of applications. PMID- 16264255 TI - The point spread function of spiral CT. AB - Computed tomography (CT) scanners are usually described by their in-plane resolution and slice-sensitivity profile (SSP). Other imaging systems are characterized by their point spread function (PSF). The PSF is an excellent basis for the analysis, design and enhancement of imaging systems. The 3D PSF of CT systems has rarely been considered, and has usually been approximated by a 3D Gaussian. We present mathematical analysis of the PSF of single-slice and multi slice fan-beam and cone-beam CT, for major reconstruction algorithms. We show that the PSF has a complicated, non-separable 3D shape. It is anisotropic in the xy plane and twisted in the z direction. Furthermore, the PSF is space variant in all three axes. In particular, it rotates as the input impulse function moves in the z direction. The PSF may also have effective discontinuities that can lead to streaking artefacts. Indirect measurements of the PSF can be misleading. We support the theoretical results by direct experimental measurements of the PSF. PMID- 16264254 TI - Resonance frequency of microbubbles in small blood vessels: a numerical study. AB - Microbubbles are currently used as ultrasound contrast agents. Their potential therapeutic applications are also under investigation. This work is designed to provide some insight into the mechanisms of energy absorption and deposition by a preformed gas bubble in the microvasculature to optimize its efficacy. In the linear regime, the most favourable condition for the transfer of energy from an ultrasonic field to a gas bubble occurs when the centre frequency of the ultrasonic field equals the resonance frequency of the bubble. The resonance frequency of gas microbubbles has been investigated up to now mainly in unbounded liquids; however when bubbles are confined in small regions, their resonance frequency is strongly affected by the surrounding boundaries. A parametric study on how the resonance frequency of microbubbles in blood vessels is affected by the bubble radius, vessel radius and the bubble position in the vessel is presented. The resonance frequency decreases below its free value with decreasing vessel radius for vessels smaller than 200-300 microm depending on the bubble size. This model suggests the possibility of using ultrasound in a range of frequencies that are, in general, lower than the ones used now for therapeutic and diagnostic applications of ultrasound (a few MHz). When microbubbles oscillate at their resonance frequency they absorb and therefore emit more energy. This energy may allow specific blood vessels to be targeted for both diagnostic and therapeutic applications of ultrasound. PMID- 16264256 TI - Simulation of realistic abnormal SPECT brain perfusion images: application in semi-quantitative analysis. AB - Simulation is useful in the validation of functional image analysis methods, particularly when considering the number of analysis techniques currently available lacking thorough validation. Problems exist with current simulation methods due to long run times or unrealistic results making it problematic to generate complete datasets. A method is presented for simulating known abnormalities within normal brain SPECT images using a measured point spread function (PSF), and incorporating a stereotactic atlas of the brain for anatomical positioning. This allows for the simulation of realistic images through the use of prior information regarding disease progression. SPECT images of cerebral perfusion have been generated consisting of a control database and a group of simulated abnormal subjects that are to be used in a UK audit of analysis methods. The abnormality is defined in the stereotactic space, then transformed to the individual subject space, convolved with a measured PSF and removed from the normal subject image. The dataset was analysed using SPM99 (Wellcome Department of Imaging Neuroscience, University College, London) and the MarsBaR volume of interest (VOI) analysis toolbox. The results were evaluated by comparison with the known ground truth. The analysis showed improvement when using a smoothing kernel equal to system resolution over the slightly larger kernel used routinely. Significant correlation was found between effective volume of a simulated abnormality and the detected size using SPM99. Improvements in VOI analysis sensitivity were found when using the region median over the region mean. The method and dataset provide an efficient methodology for use in the comparison and cross validation of semi-quantitative analysis methods in brain SPECT, and allow the optimization of analysis parameters. PMID- 16264257 TI - Transaxial system models for jPET-D4 image reconstruction. AB - A high-performance brain PET scanner, jPET-D4, which provides four-layer depth-of interaction (DOI) information, is being developed to achieve not only high spatial resolution, but also high scanner sensitivity. One technical issue to be dealt with is the data dimensions which increase in proportion to the square of the number of DOI layers. It is, therefore, difficult to apply algebraic or statistical image reconstruction methods directly to DOI-PET, though they improve image quality through accurate system modelling. The process that requires the most computational time and storage space is the calculation of the huge number of system matrix elements. The DOI compression (DOIC) method, which we have previously proposed, reduces data dimensions by a factor of 1/5. In this paper, we propose a transaxial imaging system model optimized for jPET-D4 with the DOIC method. The proposed model assumes that detector response functions (DRFs) are uniform along line-of-responses (LORs). Then each element of the system matrix is calculated as the summed intersection lengths between a pixel and sub-LORs weighted by a value from the DRF look-up-table. 2D numerical simulation results showed that the proposed model cut the calculation time by a factor of several hundred while keeping image quality, compared with the accurate system model. A 3D image reconstruction with the on-the-fly calculation of the system matrix is within the practical limitations by incorporating the proposed model and the DOIC method with one-pass accelerated iterative methods. PMID- 16264258 TI - Optimization of intensity-modulated radiation therapy with biological objectives. AB - IMRT treatment planning via biological objectives gives rise to constrained nonlinear optimization problems. We consider formulations with nonlinear objectives based on the equivalent uniform dose (EUD), with bound constraints on the beamlet weights, and describe fast, flexible variants of the two-metric gradient-projection approach for solving them efficiently and in a mathematically sound manner. We conclude that an approach that calculates the Newton component of the step iteratively, by means of the conjugate-gradient algorithm and an implicit representation of the Hessian matrix, is most effective. We also present an efficient heuristic for obtaining an approximate solution with a smoother distribution of beamlet weights. The effectiveness of the methods is verified by testing on a medium-scale clinical case. PMID- 16264259 TI - On the feasibility of water calorimetry with scanned proton radiation. AB - Water calorimetry is considered to be the most direct primary method to realize the physical quantity gray for absorbed dose to water. The Swiss Federal Office of Metrology and Accreditation (METAS) has routinely operated a water calorimeter as primary standard for photon radiation since 2001. Nowadays, cancer therapy with proton radiation has become increasingly important and is a well established method. In the framework of the ProScan project conducted by the Paul Scherrer Institute (PSI), the spot-scanning technique is prepared for the subsequent application in hospitals, and adjusted to the recent findings of clinical research. In the absence of primary standards for proton radiation, the metrological traceability is assured by calibrating secondary standards in 60Co radiation and correcting with calculated beam quality correction factors. It is internationally recognized that the development of primary standards for proton radiation is highly desirable. In a common project of PSI and METAS, it is investigated whether a modified version of the water calorimeter in operation at METAS is suitable as primary standard for scanned proton radiation. A feasibility study has been conducted to investigate the linear energy transfer (LET) dependence of the heat defect and the influence of the time and space structure of the scanned beam on the homogeneity and stability of the temperature field in the water calorimeter. Simulations are validated against experimental data of the existing calorimeter used with photon radiation and extended to scanned proton radiation. PMID- 16264261 TI - Observations of a human hair shaft with an x-ray microscope. AB - We observed the internal structures of a human hair shaft using x-ray microscopes with a spatial resolution in the range from a few microns to less than 100 nm. The energy of the x-ray used is 6.95 keV. The Zernike phase contrast together with a spatial resolution better than 100 nm enabled us to see the cuticles of scales, the cortex of macrofibrils and the medulla. All these internal features and more can easily be observed with no sample preparation including staining. PMID- 16264260 TI - Simulation of small-angle x-ray scattering from collagen fibrils and comparison with experimental patterns. AB - Simulation of small-angle x-ray scattering from collagen in healthy and cancerous breast tissue may reveal detailed information on the structural changes in collagen. Collagen fibril is modelled as a cylinder with axially periodic step function electron density, and packing is approximated by placing the cylinders in small hexagonal bundles. The intensity from a bundle is calculated by summing analytical scattering amplitudes from the cylinders, and intensities from several bundles with varying lattice constants are averaged. Comparisons with more complex models are made to estimate the robustness of the model. The oscillations in the equatorial direction are not significantly affected by added complexity. The relative intensities of the Bragg peaks in the meridional direction can be tuned by modifying the axial electron density distribution. Tests with different fibril radius distributions show that the average radius can be determined with an accuracy of +/-0.5 nm but that the shape of the radius distribution cannot be accurately determined from the scattering patterns. The effect of multiple scattering and the detector point-spread function (PSF) is considered, and the PSF may make a significant contribution to the final slope of the scattering pattern. Comparisons with observed scattering indicate that the model is basically correct at the supra-molecular level. PMID- 16264262 TI - Characterization of newly established oral cancer cell lines derived from six squamous cell carcinoma and two mucoepidermoid carcinoma cells. AB - Since genetic abnormalities of human cancer are greatly geographically dependent, cultural and environmental backgrounds are thought to be closely related to the carcinogenic process. In the present study, eight human cell lines were established by culture from untreated carcinomas of the oral cancer, of which five were from primary oral squamous cell carcinomas (OSC), one from a mucoepidermoid carcinoma (MEC) and one each originating from metastatic OSC and MEC. All the studied tumor lines grew as monolayers, and showed: i) an epithelial origin by the presence of cytokeratin, and ii) tumorigenic potential in nude mice. Western blot analysis revealed i) over expression of EGFR in six of the cell lines ii) decreased expression of E-cadherin in six cell lines compared to normal human oral mucosa. A mutational analysis showed: point mutations of p53 at exon 7, with transversion, and at exon 8, with transition. These well characterized human YD cell lines should serve as useful tools in the study of the molecular pathogenesis and biological characteristics of head and neck cancer cells, and in the future testing of new therapeutic reagents for oral cancer. PMID- 16264263 TI - PTEN/MMAC1 enhances the growth inhibition by anticancer drugs with downregulation of IGF-II expression in gastric cancer cells. AB - PTEN/MMAC1 is a tumor suppressor gene that is mutated in a variety of advanced and metastatic cancers. Its major function is likely to be the phosphatase activity that regulates the phosphotidylinositol (PI)3-kinase/Akt pathway. On the other hand, IGF system plays an important role in cell proliferation and cell survival via PI3-kinase/Akt and mitogen-activated protein kinase pathways in many cancer cells. To evaluate effect of PTEN on cell growth and IGF system in gastric cancer, human gastric adenocarcinoma cells (SNU-5 & -216) were transfected with human PTEN cDNA. Those PTEN- transfected gastric cancer cells had a lower proliferation rate than the pcDNA3-transfected cells. PTEN overexpression induced a profound decrease in the IGF-II and IGF-IR expression levels, and downregulation of IGF-II expression by PTEN was mediated through the regulation of the IGF-II promoter. In addition, a PI3-kinase inhibitor, LY294002, induced the downregulation of IGF-II expression. The PTEN-overexpressing SUN-5 and -216 cells were more sensitive to death induced by etoposide and adriamycin that induce DNA damage than the pcDNA3-transfected cells. These findings suggest that PTEN suppresses the cell growth through modulation of IGF system and sensitizing cancer cells to cell death by anticancer drugs. PMID- 16264264 TI - Efficient culture system for human embryonic stem cells using autologous human embryonic stem cell-derived feeder cells. AB - Human embryonic stem cells (hESCs) need feeder cells for their maintenance in an undifferentiated state. In conventional culture systems, mouse embryonic fibroblasts (MEFs) serve as feeder cells to maintain hESCs. However, the use of MEFs elevates the risk of transmitting mouse pathogens and thus limits the potential of hESCs in cell replacement therapy. Consequently, the use of human feeder cells would be an important step forward in this in vitro technology. To address this issue, we used fibroblast-like cells differentiated from the Miz hES6 hESC line (DiffMiz-hES6) as feeder cells to support the in vitro growth of three hESC lines. Immunofluorescence microscopy and reverse transcription-PCR assessing the expression of undifferentiated hESC markers revealed all three hESC lines were maintained in an undifferentiated state. In vitro proliferation proceeded as efficiently as when the hESCs were cultured on MEFS. Moreover, karyotype analysis revealed the chromosomal normality of the hESC lines and the DiffMiz-hES6 feeders themselves after even 50 passages. Furthermore, the hESC lines maintained their pluripotency since they remained capable of forming embryoid bodies (EBs) in vitro. Thus, hESC-derived fibroblast-like cells successfully support in vitro hESC propagation. PMID- 16264265 TI - Roles of TNF-alpha and IgE in the late phase of contact hypersensitivity induced by trimellitic anhydride. AB - Trimellitic anhydride (TMA) is widely used industrially to make epoxy and alkyd resins, plasticizers and surfactants. The purpose of this study was to investigate whether contact hypersensitivity (CHS) is induced by repeated TMA challenge and the role of TNF-alpha and IgE in the TMA-induced CHS. The repetition of the challenge enlarged the extent of an early and a late phase of CHS in TNF-alpha+/+ (B6129SF2/J) and Balb/c mice. In the late phase of TMA induced CHS, the peak of ear swelling responses by single challenge showed at 24 h after challenge, but the peak was observed at 8 h after repeated challenge. In the TNF-alpha knockout TNF-alpha-/- (B6;129S-Tnftm1Gk1) mice, the repetition of the TMA challenges enlarged the extent of the late phase of CHS, but less than those in TNF-alpha+/+ mice. Injection of anti-TNF-alpha antibody into the peritoneal cavity of Balb/c mice significantly decreased the extent of the late phase of CHS. Subcutaneous injection of anti-IgE antibody into Balb/c mice also decreased the extent of the late phase of CHS in dose-dependent manner. Histologically, infiltration of polymorphonuclear leukocytes and eosinophils was more pronounced in repeatedly TMA-challenged TNF-alpha+/+ and Balb/c mice than in the TNF-alpha-/- mice and anti-TNF-alpha or anti-IgE antibodies treated Balb/c mice. These results indicate that mice sensitized by TMA could possibly offer a useful model to study the mechanism of CHS, and TNF-alpha and IgE may act as potential modulators in the late phase of TMA-induced CHS. Neutralization of TNF alpha and IgE by anti-TNF-alpha or anti-IgE antibodies may provide therapeutic tools for the treatment of TMA-induced CHS. PMID- 16264266 TI - Involvement of protein kinase C pathway in UVC-stimulated phospholipase D2 activity in Vero 76 cells. AB - Phospholipase D (PLD) activity is known to be related to oxidant-induced cellular signaling and membrane disturbance. Previously, an induction of PLD activity in various cell lines by X-ray irradiation was observed. In this study, we examined the effect of UVC radiation on the PLD activity in Vero 76 cells. At a dose of 10 kJ/m2 of UVC irradiation, the PLD activity was stimulated approximately 10-fold over the basal activity. This UVC-induced PLD activity was found to be dependent on the presence of extracellular calcium and was inhibited by catalase as well as amifostine-an intracellular thiol antioxidant. Pretreatments with Ro32-0432-a selective inhibitor of protein kinase C (PKC)-and downregulation of PKC by preincubation of phorbol 12-myristate 13-acetate significantly inhibited the UVC induced PLD activity. UVC-stimulated PLD activity was observed only in murine PLD2 (mPLD2)-transfected Vero 76 cells and not in human PLD1 (hPLD1)-transfected cells. Transient incorporation of PKC with mPLD2 and the phosphorylation of mPLD2 by a and b forms of PKC by UVC irradiation were observed. These results suggest that the UVC-stimulated PLD activity in Vero 76 cells is mediated through transient phosphorylation of PLD2 by the translocation of PKC to PLD2. PMID- 16264267 TI - Heat shock protein 27 interacts with vimentin and prevents insolubilization of vimentin subunits induced by cadmium. AB - Vimentin is an intermediate filament that regulates cell attachment and subcellular organization. In this study, vimentin filaments were morphologically altered, and its soluble subunits were rapidly reduced via cadmium chloride treatment. Cadmium chloride stimulated three major mitogen-activated protein kinases (MAPKs): extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38, and led apoptotic pathway via caspase-9 and caspase-3 activations. In order to determine whether MAPKs were involved in this cadmium induced soluble vimentin disappearance, we applied MAPK-specific inhibitors (PD98059, SP600125, SB203580). These inhibitors did not abolish the cadmium induced soluble vimentin disappearance. Caspase and proteosome degradation pathway were also not involved in soluble vimentin disappearance. When we observed vimentin levels in soluble and insoluble fractions, soluble vimentin subunits shifted to an insoluble fraction. As we discovered that heat-shock protein 27 (HSP27) was colocalized and physically associated with vimentin in unstressed cells, the roles of HSP27 with regard to vimentin were assessed. HSP27 overexpressing cells prevented morphological alterations of the vimentin filaments, as well as reductions of soluble vimentin, in the cadmium-treated cells. Moreover, HSP27 antisense oligonucleotide augmented these cadmium-induced changes in vimentin. These findings indicate that HSP27 prevents disruption of the vimentin intermediate filament networks and soluble vimentin disappearance, by virtue of its physical interaction with vimentin in cadmium-treated SK-N-SH cells. PMID- 16264269 TI - Cardiac expression profiles of the naked DNA vectors encoding vascular endothelial growth factor and basic fibroblast growth factor. AB - We investigated expression profiles and biological effects of the naked DNA vectors in the heart. To this end, naked DNA vector was injected into the apex of the beating rat heart after thorocotomy. When the expression of LacZ reporter was examined by reverse transcription-PCR and histochemical staining for beta galactosidase, LacZ expression was detected only in the heart, suggesting limited dissemination of the injected vector in vivo. Even within the heart, LacZ expression was limited to the injection area (apex). Similar observations were made with other transgenes such as VEGF and basic fibroblast growth factor (bFGF), where 77% and 69% of the total transgene exprssion were detected in the heart segments containing the apex. Although VEGF and bFGF expressions were detected until 2 weeks after DNA injection, the highest levels of VEGF and bFGF were observed on day 5 and day 1, respectively. The optimal doses of the vectors were 10 microg and 25 microg for the VEGF and bFGF vectors, respectively. Interestingly, injection of bFGF vector led to 50% increase in the level of endogenous murine VEGF expression. Consistent with this finding, the number of vessels that stained positive for alpha-smooth muscle actin was increased in the bFGF vector-injected heart. These results suggest that simple injection of naked DNA vector may be sufficient to induce significant angiogenesis in the myocardium and that naked DNA gene therapy may be a feasible approach for the treatment of ischemic heart disease. PMID- 16264268 TI - Natural compounds,fraxin and chemicals structurally related to fraxin protect cells from oxidative stress. AB - Coumarins comprise a group of natural phenolic compounds found in a variety of plant sources. In view of the established low toxicity, relative cheapness, presence in the diet and occurrence in various herbal remedies of coumarins, it appears prudent to evaluate their properties and applications further. The purpose of this study is to investigate cellular protective activity of coumarin compound, fraxin extracted from Weigela florida var. glabbra, under oxidative stress, to identify genes expressed differentially by fraxin and to compare antioxidative effect of fraxin with its structurally related chemicals. Of the coumarins, protective effects of fraxin against cytotoxicity induced by H2O2 were examined in human umbilical vein endothelial cells (HUVECs). Fraxin showed free radical scavenging effect at high concentration (0.5 mM) and cell protective effect against H2O2-mediated oxidative stress. Fraxin recovered viability of HUVECs damaged by H2O2-treatment and reduced the lipid peroxidation and the internal reactive oxygen species level elevated by H2O2 treatment. Differential display reverse transcription-PCR revealed that fraxin upregulated antiapoptotic genes (clusterin and apoptosis inhibitor 5) and tumor suppressor gene (ST13). Based on structural similarity comparing with fraxin, seven chemicals, fraxidin methyl ether (29.4% enhancement of viability), prenyletin (26.4%), methoxsalen (20.8%), diffratic acid (19.9%), rutoside (19.1%), xanthyletin (18.4%), and kuhlmannin (18.2%), enhanced more potent cell viability in the order in comparison with fraxin, which showed only 9.3% enhancement of cell viability. These results suggest that fraxin and fraxin-related chemicals protect HUVECs from oxidative stress. PMID- 16264270 TI - The extracellular calcium sensing receptor is expressed in mouse mesangial cells and modulates cell proliferation. AB - The extracellular calcium sensing receptor (CaSR) belongs to the type III family of G-protein-coupled receptors, a family that comprises the metabotropic glutamate receptor and the putative vomeronasal organ receptors. The CaSR plays an important role for calcium homeostasis in parathyroid cells, kidney cells and other cells to directly 'sense' changes in the extracellular calcium ion concentration ([Ca2+]o). The mesangial cells are known to be involved in many pathologic sequences through the mediation of altered glomerular hemodynamics, cell proliferation, and matrix production. In this study, we examined the expression of the CaSR in the mouse mesangial cell lines (MMC, ATCC number CRL 1927). Reverse transcription-polymerase chain reaction (RT-PCR) was perform with CaSR-specific primers, and this was followed by nucleotide sequencing of the amplified product; this process identified the CaSR transcript in the MMCs. Moreover, CaSR protein was present in the MMCs as assessed by Western blot and immunocytochemical analysis using a polyclonal antibody specific for the CaSR. Functionally, [Ca2+]o induced the increment of the intracellular calcium concentration ([Ca2+]i) in a dose-dependent manner. This [Ca2+]i increment by [Ca2+]o was attenuated by the pretreatment with a phospholipase C inhibitor (U73122) and also by a pretreatment with a CaSR antagonist (NPS 2390). The similar results were also obtained in IP3 accumulation by [Ca2+]o. To investigate the physiological effect of the CaSR, the effect of the [Ca2+]o on cell proliferation was studied. The increased [Ca2+]o (up to 10 mM) produced a significant increase in the cell numbers. This mitogenic effect of [Ca2+]o was inhibited by the co-treatment with a CaSR antagonist. From these results, the [Ca2+]o-induced [Ca2+]i elevation in the MMC is coupled with the extracellular calcium sensing receptor. Furthermore, [Ca2+]o produces a mitogenic effect in MMCs. PMID- 16264271 TI - Menin represses JunD transcriptional activity in protein kinase C theta-mediated Nur77 expression. AB - TCR signaling leading to thymocyte apoptosis is mediated through the expression of the Nur77 family of orphan nuclear receptors. It has been shown that the Nur77 promoter is activated by at least two signaling pathways, one mediated by calcium and the other by protein kinase C (PKC). MEF2D has been known to regulate Nur77 expression in a calcium- dependent manner. The mechanism by which calcium regulates MEF2D is through dissociation of calcium-sensitive MEF2 corepressors (Cabin1/HDACs, HDAC4/5) and the association with calcineurin-activated transcription factor NF-AT and the coactivator p300. However, little is known about how PKC activates the Nur77 promoter. Herein, we report that PKCtheta targets AP-1 like response element in the Nur77 promoter where JunD constitutively binds. PKCtheta triggers mitogen-activated protein kinase inediated phosphorylation of JunD, and increases transcriptional activity of JunD, cooperatively with p300. Menin is identified as the transcriptional corepressor for JunD via recruitment of mSin3-istone deacetylases. In fact, Menin represses PKCtheta/p300-mediated transcriptional activity of JunD in T cell. Its dynamic regulation of histone modifiers with JunD is responsible for PKCq synergistic effect on Nur77 expression in T cell. PMID- 16264272 TI - Alternative therapy of earth elements increases the chondroprotective effects of chondroitin sulfate in mice. AB - The administration of mineral sulphur water is an alternative experimental approach for the treatment of rheumatic diseases, such as osteoarthritis (OA), that cause the degeneration of bone and cartilage and sufferance to the patients. Chondroitin sulfate (CS) is a symptomatic slow acting nutropeucital agent currently used in molecular therapy of OA. Therefore, we have studied the role and efficacy of the selective soil paste from the mineral sulphur enriched spring (mud)-therapy alone or in combination with CS in the treatment of OA. The study was performed on 40 C57 Black 6N mice, an experimental model which spontaneously develop an osteoarthritic process. The animals were divided in 4 groups and were treated with the single agents or with the combination. After 30 days of treatment all the mice were sacrificed and right knees and blood were collected. It was found that CS determined a reduction of radiological and histological features of chondrodegeneration and that mud-therapy increased the effects of CS in the animal group treated with the combination. However, the effects of thermal therapy alone were not statistically significant. Since OA is characterized by an increase of the production of nitric oxide (NO) by chondrocytes in extracellular matrix with its consequent elevation in serum and synovial fluid, we have evaluated the effects of the treatments on serum NO levels. CS alone induced a statistically significant reduction of NO serum levels (90+/-13 micromM vs 219+/ 60 microM of control group, P<0.05) while mud-therapy alone induced a not statistically significant reduction of serum NO (170+/-62 microM, P>0.05). However, the latter strongly potentiated the decrease of serum NO induced by CS (31+/-1.5 microM) with a high statistical significance if compared to both the control group (P<0.01) and the CS-treated group (P<0.05). In conclusion, this study demonstrates that mud-therapy with sulphur mineral water could represent an important phase of the therapeutic strategy of OA. This experimental strategy could integrate and potentiate the standard pharmacological tools. Moreover, we have set a valid experimental in vivo model for the study of the thermal effects on the development of OA. PMID- 16264273 TI - Centrosome amplification and multinuclear phenotypes are Induced by hydrogen peroxide. AB - Multinucleated cells resulted from mitosis defect have been noted in pathophysiological states of the cells such as inflammation, senescence and cancer. Since oxidative stress has been known to correlate with these pathophysiological conditions, we tested the effect of H2O2 on the cell cycle progression and formation of multinucleated cells. H2O2 induced a significant delay in cell cycle progression in Chang liver cells. Interestingly, H2O2 actively induced hyperamplification of centrosomes (n>or=3) and multipolar spindle formation during mitosis and subsequently increased the generation of multinucleated cells. A significant increase of the phospho-ERK level was observed upon H2O2 treatment but PD98059, an MEK1/2 inhibitor, didn't reduce the frequency of cells with hyperamplified centrosomes. On the other hand, treatment of either H2O2 or adriamycin increased intracellular ROS levels and multinucleated cells, which were significantly suppressed by antioxidants, N acetylcysteine and PDTC. Thus, our results suggest that oxidative stress can trigger centrosome hyperamplification and multinucleated cell formation, which may promote pathophysiological progression. PMID- 16264274 TI - Transcription repression of a CCAAT-binding transcription factor CBF/HSP70 by p53. AB - NF-Y transcription factor binds to CCAAT boxes on promoters of cell cycle regulatory genes such as cdc2, cyclin B, cdc25C, and cyclin A. We previously reported that the DNA binding activity of NF-Y is regulated by p53-p21-cdk2 pathway. CBF/HSP70 was originally identified as a transcription factor binding to the CCAAT box on the hsp70 promoter and mediates transcription repression of hsp70 pro- moter by p53. Recently it was demonstrated that CBF/HSP70 interacts and cooperates with NF-Y. In this study, we found that p53 represses the trans cription of CBF/HSP70. Since transactivation ability of NF-Y is regulated in a cell cycle-dependent manner, we examined the transcription of CBF/HSP70 during the cell cycle. After synchronization of a human bladder carcinoma cell lacking functional p53 at early S phase, we infect the cells with adenovirus encoding p53. Cells infected with control virus progressed to S and G2 after release from the arrest. In contrast, cells expressing p53 enter S and G2 phases, but arrest at G2/M. The expression of CBF/HSP70 was induced at S/G2 phase in cells infected with a control virus, but kept to be repressed in cells expressing p53. Thus, these results suggest that p53 suppresses the expression of cell cycle regulatory genes though inhibiting both CCAAT binding factors, CBF/HSP70 and NF-Y. PMID- 16264275 TI - Coordinated transcriptional regulation of calmegin,a testis-specific molecular chaperon, by histone deacetylase and CpG methyltransferase. AB - Calmegin is a testis-specific molecular chaperon playing a key role in spermatogenesis. However, the transcriptional regulatory mechanisms for calmegin expression are entirely unknown. Herein, we revealed that calmegin is transcriptionally regulated by histone deacetylase (HDAC) and CpG methyltransferase. The cDNA microarray analysis of the human fibrosarcoma cells treated with trichostatin A (TSA) showed an increased level of calmegin mRNA. The induction of calmegin mRNA by TSA was added by the treatment with 5-aza-2' deoxycytidine (5'Aza-dC), implying that epigenetic alterations are involved in the transcriptional repression of the gene. Moreover, chromatin immunoprecipitation assay using an anti-acetyl-histone H3 antibody exhibited that the proximal region (-152 ~ -31) of the calmegin promoter is responsible for HDAC mediated transcriptional repression of the gene. These results demonstrate that calmegin expression is regulated by HDAC and CpG methyltransferase in a coordinative way. PMID- 16264276 TI - Up-regulation of defense enzymes is responsible for low reactive oxygen species in malignant prostate cancer cells. AB - Reactive oxygen species (ROS) are involved in a diversity of important phenomena in the process of tumor development. To investigate the alterations of oxidative stress and their related systems in tumor progression, a variety of components in the antioxidative stress defense system were examined in prostate cancer cell lines, PC3 and LNCaP. Cell surface molecules involved in metastasis were expressed highly in PC3 cells compared with LNCaP cells, and strong invasion ability was shown in PC3 cells only. ROS level in LNCaP cells was twice higher than that in PC3 cells, although nitric oxide (NO) level was similar between the two cell lines. The content of GSH increased up to about 2-fold in PC3 compared with LNCaP. Activities of glutathione reductase, thioredoxin reductase, and glutathione S-transferase except catalase are significantly higher in PC3 cells than in LNCaP cells. Furthermore, oxidative stress-inducing agents caused down regulation of GSH and glutathione S-transferase much more significantly in LNCaP cells than in PC3 cells. These results imply that malignant tumor cells may maintain low ROS content by preserving relatively high anti-oxidative capacity, even in the presence of stressful agents. PMID- 16264277 TI - Detection of single nucleotide insertion of BCR/ABL region in imatinib-resistant human myelogenous leukemia SR-1 cells. AB - Imatinib mesylate is a selective Bcr/Abl kinase inhibitor and an effective anticancer agent for Bcr/Abl-positive chronic myelogenous leukemia. Most patients in chronic phase maintain durable responses; however, many in blast crisis fail to respond, or relapse quickly. Mutations within the BCR/ABL kinase domain are the most commonly identified mechanism associated with relapse. To overcome the imatinib resistance in CML, many investigators have tried to clarify molecular mechanism for imatinib resistance in cells of patients who failed to respond to imatinib. Our aim was to invesitigate underlying mechanism for imatinib resistance in SR-1 cells, which were derived from a CML patient in blast crisis. We detected the new mutation of BCR/ABL, resulting in premature termination and loss of BCR/ABL fusion protein expression, which might be possible mechanism for the resistance to imatinib in SR-1 cells. PMID- 16264278 TI - Water, water everywhere: dare we drink? We don't have to go to New Orleans to find contamination. PMID- 16264279 TI - Not enough nurses, education, and technology: increasing staffing in nursing homes may not be enough. PMID- 16264280 TI - Retirement? PMID- 16264281 TI - Keep dialogue alive. PMID- 16264282 TI - Keep dialogue alive. PMID- 16264283 TI - Keep dialogue alive. PMID- 16264284 TI - Keep dialogue alive. PMID- 16264285 TI - Ethnopharmacology. PMID- 16264286 TI - Nurses and HIV. PMID- 16264287 TI - Safe career. PMID- 16264288 TI - Angels? PMID- 16264289 TI - Angels? PMID- 16264290 TI - Angels? PMID- 16264291 TI - 'Negligent homicide' charges for nursing home owners: the frailest residents paid the highest price when Katrina struck. PMID- 16264292 TI - You get what you pay for: news flash: higher nurse-patient ratios still save lives. PMID- 16264295 TI - World health roundup. PMID- 16264294 TI - Women's health research: evidence from two major trials. PMID- 16264296 TI - On the road to disaster--and back: AJN's news director visited health officials and rescuers immediately after Hurricane Katrina. PMID- 16264298 TI - To give blood or not to give: should there be a question? PMID- 16264299 TI - Solutions to RN staffing: Texas and Oregon are models for other states addressing the crisis. PMID- 16264300 TI - Legionella pneumonia: many cases of Legionnaire disease go unreported or unrecognized. PMID- 16264301 TI - The other cancer story: is this a breast cancer competition? PMID- 16264302 TI - Tainted water on tap: what to tell patients about preventing illness from drinking water. AB - Annual cases of waterborne illness in the United States are estimated to number about 900,000, but most experts believe the incidence to be much higher. The U.S. Environmental Protection Agency regulates the nation's drinking water supply, setting maximum allowable levels for 87 known natural and synthetic contaminants; but thousands more go unregulated. This article describes selected contaminants and their known health effects, which range from acute gastroenteritis to cancer and reproductive and developmental effects. It discusses which populations are more vulnerable, outlines assessment, and elucidates nurses' roles in patient education and as community advocates for safer drinking water. PMID- 16264304 TI - Acute pancreatitis: diagnosis and treatment of a potentially fatal condition. PMID- 16264305 TI - RN staffing time and outcomes of long-stay nursing home residents: pressure ulcers and other adverse outcomes are less likely as RNs spend more time on direct patient care. AB - OBJECTIVE: A clear link has been demonstrated between lower nurse staffing levels in hospitals and adverse patient outcomes, but the results of studies of such relationships in long-term care facilities haven't been as clear. This study explored the time nurses spent in direct care and how it affected outcomes in long-stay (two weeks or longer) nursing home residents. METHODS: In a retrospective study of data collected as part of the National Pressure Ulcer Long Term Study (NPULS), we analyzed data on 1,376 residents of 82 long-term care facilities whose lengths of stay were 14 days or longer, who were at risk of developing pressure ulcers but had none at study entry, and who had a Braden Scale score of 17 or less. Primary data came from residents' medical records during 12-week periods in 1996 and 1997. Dependent variables included development of pressure ulcer or urinary tract infection (UTI), weight loss, deterioration in the ability to perform activities of daily living (ADLs), and hospitalization. Independent variables included resident demographics, severity of illness, nutritional and incontinence interventions, medications, and nurse staffing time. RESULTS: More RN direct care time per resident per day (examined in 10-minute increments up to 30 to 40 minutes per resident per day) was associated with fewer pressure ulcers, hospitalizations, and UTIs; less weight loss, catheterization, and deterioration in the ability to perform ADLs; and greater use of oral standard medical nutritional supplements. More certified nursing assistant and licensed practical nurse time was associated with fewer pressure ulcers but did not improve other outcomes. CONCLUSIONS: The researchers controlled for important variables in long-stay nursing home residents at risk for pressure ulcers and found that more RN direct care time per resident per day was strongly associated with better outcomes. There's an urgent need for further research to confirm these findings and, if confirmed, for improving RN staffing in nursing homes to decrease avoidable adverse outcomes and suffering. PMID- 16264316 TI - The battle over RN replacement: the ANA and nurses take action to protect the public by maintaining nursing services. PMID- 16264317 TI - Medication reconciliation: verifying medication orders and clarifying discrepancies should be standard practice. PMID- 16264319 TI - Challenging how competency is validated: the wound, ostomy and continence nurses society issues a position statement. PMID- 16264320 TI - The 2005 AJN-Beatrice Renfield: caring for the caregiver award. PMID- 16264322 TI - Gold out of seaweed: a tenacious public health nurse on the Pacific Coast. PMID- 16264323 TI - Long-term commitment and change: one nurse focuses on nursing home reform. PMID- 16264324 TI - Every patient deserves a safe nurse: patient safety cannot be achieved when nurses are not safe. PMID- 16264326 TI - WASP and the phenotypic range associated with deficiency. AB - PURPOSE OF REVIEW: This review reports on the range of clinical phenotypes that are caused by mutations in the Wiskott-Aldrich Syndrome Protein (WASP) gene. The basis of genotype-phenotype correlation in Wiskott-Aldrich syndrome (WAS) is discussed, with regard to expression of the WAS protein (WASp) and of the effects of WASP mutations on WASp function. Advances in preclinical models of gene therapy for WAS are presented. RECENT FINDINGS: Two recent studies have supported genotype-phenotype correlation in WAS and in related X-linked thrombocytopenia. Expression of the WASp was found to be the best predictor of clinical phenotype. Investigation of autoimmune manifestations associated with WAS has shown that autoimmune hemolytic anemia and elevated serum IgM associate with a more severe clinical course. Finally, while results of hematopoietic stem cell transplantation for WAS continue to improve, several studies have shown the potential benefit of novel therapeutic approaches based on gene transfer. In particular, use of lentiviral vector-driven expression of the WASP gene under autologous promoter sequences has been found to result in increased targeting of hematopoietic stem cells, higher levels of WASp expression, and improved reconstitution of immune function. SUMMARY: Availability of tools that allow analysis of WASp expression has provided evidence for a genotype-phenotype correlation in patients with WASP gene defects. Protein expression is an important prognostic indicator. The molecular and cellular abnormalities of WAS associated defects are being identified, and significant advances in vector mediated gene transfer have opened possibilities for the treatment of WAS based on gene therapy. PMID- 16264327 TI - CD3 deficiencies. AB - PURPOSE OF REVIEW: The molecular characterization of inherited T-cell immunodeficiencies has contributed to delineating key factors in human T-cell development. This review reports on the recent description of deleterious mutations in the genes encoding CD3 subunits expressed at the T-lymphocyte membrane in association with the T-cell receptor. RECENT FINDINGS: Homozygous mutations in CD3D and CD3E genes lead to a complete block in T-cell development and thus to an early-onset severe combined immunodeficiency phenotype. Thymic studies have shown that the defect in T-cell development occurs at the transition between 'double-negative' and 'double-positive' thymocytes. These results contrast with the partial T-cell immunodeficiency caused by a deficiency in CD3G. SUMMARY: Two new severe combined immunodeficiency conditions have been reported as a consequence of either CD3D or CD3E deficiency. The distinct phenotype of CD3G deficiency sheds light on the differential roles of CD3 subunits in T lymphocyte development. PMID- 16264329 TI - Tackling the heterogeneity of CVID. AB - PURPOSE OF REVIEW: Common variable immunodeficiency is clinically the most relevant primary immunodeficiency of the adult. Its heterogeneity has hindered progress in the pathogenetic understanding of the majority of common variable immunodeficiency patients. This abstract summarizes recent aspects of the field and emphasizes the need for a commonly accepted approach to classify common variable immunodeficiency. RECENT FINDINGS: In the last 2 years, the first genetic defects underlying common variable immunodeficiency, including ICOS, TACI, BAFF-R and CD19, have been identified. The analysis of dendritic cells demonstrated alterations in a majority of patients in addition to the disturbed T and B-cell function. Several changes of the adaptive immune system might be secondary to an underlying chronic inflammatory setting possibly due to a HHV8 infection in a subgroup of patients with granulomatous disease, autoimmune phenomena and T-cell dysfunction. The occurrence of granulomatous inflammation is associated with a worse prognosis compared with common variable immunodeficiency patients without granuloma. SUMMARY: The pathogenesis of common variable immunodeficiency includes disturbances of the adaptive as well as innate immune system. Identified monogenic defects account for about 10% of cases, leaving the majority of defects undefined and certainly in part epigenetic. To combine the known aspects of the pathogenesis of common variable immunodeficiency to a conclusive picture, the clinical and immunologic phenotyping of patients needs to be standardized. PMID- 16264328 TI - TACItly changing tunes: farewell to a yin and yang of BAFF receptor and TACI in humoral immunity? New genetic defects in common variable immunodeficiency. AB - PURPOSE OF REVIEW: The complex system of the tumour necrosis factor ligands BAFF and APRIL and their three receptors BCMA, TACI and BAFF receptor and its role in B-cell development and function is the objective of extensive research. Whereas the importance of BAFF/BAFF receptor interactions for B-cell survival could be clearly demonstrated, TACI is believed to counteract BAFF activity as a negative regulator in the murine model. The primarily immunodeficient phenotype of human TACI deficiency, however, claims a distinct function of this receptor in human peripheral B-cell development, class switch recombination and terminal differentiation. RECENT FINDINGS: Common variable immunodeficiency comprises a heterogeneous group of antibody deficiency syndromes characterized by impaired terminal B-cell differentiation. By means of molecular genetics common variable immunodeficiency is still ill-defined, but the description of the deficiency of the inducible costimulator in a small subgroup of common variable immunodeficiency patients set the starting point for the molecular dissection of this disease entity. The recent discovery of genetic defects in the tumour necrosis factor receptor superfamily members TACI and BAFF receptor in patients with common variable immunodeficiency denotes further advances in this field. SUMMARY: In this review we will discuss recent progress made in the understanding of the BAFF/APRIL-TACI/BCMA/BAFF receptor system in relation to the recent discovery that mutations in human TACI cause a primary humoral immunodeficiency. This suggests a refined role for TACI in human B-cell biology. PMID- 16264330 TI - Update on X-linked hypogammaglobulinemia with isolated growth hormone deficiency. AB - PURPOSE OF REVIEW: To provide an update on the syndrome X-linked hypogammaglobulinemia with isolated growth hormone deficiency, focusing on the pedigree described originally. RECENT FINDINGS: An additional case of X-linked hypogammaglobulinemia with isolated growth hormone deficiency and an unaffected male have been born to a female carrier in the family, allowing improved disease locus mapping. Unpublished research has identified a mutation in the transcription factor myeloid elf-1-like factor that may be the cause of the disease. SUMMARY: X-linked hypogammaglobulinemia with isolated growth hormone deficiency is not caused by Bruton's tyrosine kinase mutations in the family described originally, but may be due to a mutation in myeloid elf-1-like factor. PMID- 16264331 TI - The range of defects associated with nuclear factor kappaB essential modulator. AB - PURPOSE OF REVIEW: Impaired ability to signal and activate specific gene transcription through nuclear factor kappaB (NFkappaB) has been directly linked to immunodeficiency. Hypomorphic mutations in the gene encoding NFkappaB essential modulator (NEMO), located on the X chromosome, impair NFkappaB function and lead to ectodermal dysplasia with immunodeficiency (ED-ID) with increased susceptibility to pyogenic bacteria, viruses and nonpathogenic mycobacterial infections. This is due to impaired, but not abolished, response to a variety of stimuli including Toll-like receptor agonists. Alternatively, loss-of-function (amorphic) mutations in the same gene lead to incontinentia pigmenti. The purpose of this review is to explore the range of immunologic defects associated with mutations in NEMO, a key regulatory molecule in the NFkappaB pathway. RECENT FINDINGS: In addition to the discovery of X-linked recessive hypomorphic mutations in NEMO as the cause of anhidrotic ED-ID, autosomal-dominant hypermorphic mutations in inhibitor of NFkappaB (IkappaB) alpha have been described recently. In addition, a better understanding of genotype-phenotype correlation in ED-ID patients is evolving. SUMMARY: ED-ID is a combined, variable but profound immunodeficiency characterized by susceptibility to pyogenic bacteria and mycobacterial infection. Understanding the features of particular NEMO mutations will provide insight into the role of this gene and will help define the crucial role of the function and regulation of NFkappaB in the immune response. PMID- 16264332 TI - Cytokine-mediated signalling and early defects in lymphoid development. AB - PURPOSE OF REVIEW: The aim of the review is to report on recent advances in cytokine-mediated signalling, as illustrated by the study of natural human mutants. In particular, the role of cytokines and cytokine-mediated signalling in human T-cell development is analysed in detail, and currently available forms of treatment including experimental trials are described. RECENT FINDINGS: Defects of the cytokine/JAK/STAT axis have been recently described as responsible for human Severe Combined Immune Deficiency. In particular, defects in gammac, JAK3 and IL7RA have been analysed in terms of development of novel diagnostic tools as well as of new therapeutic agents for the treatment of autoimmune diseases and graft-versus-host disease. SUMMARY: Dissection of the genetic defects underlying the various forms of Severe Combined Immune Deficiency has helped develop new and more accurate diagnostic assays and novel forms of treatment. PMID- 16264333 TI - New insights into the mechanisms of allergen-specific immunotherapy. AB - PURPOSE OF REVIEW: Specific immunotherapy represents the only curative treatment of a specific allergy, and is therefore of great interest in terms of immunological mechanisms and therapeutic developments. RECENT FINDINGS: Allergen specific regulatory T cells are induced after the initiation of specific immunotherapy, and are assumed to suppress effector T cells directly mediating allergic inflammation. Therefore regulatory T cells may play a key role in the re induction of allergen tolerance. Multiple pathways in many systems are described to promote or enhance regulatory T cells. This review summarizes the current view on immunological mechanisms leading to and coming from regulatory T cells. SUMMARY: The highlighted mechanisms may not only contribute to improved specific immunotherapy, but also give insight into a clinically relevant therapy targeting regulatory T cells. The approach of addressing endogenous regulatory mechanisms specifically controlling inflammation instead of targeting inflammation itself is relevant for future pharmacological developments. PMID- 16264334 TI - Regulatory T cells as a target for induction of immune tolerance in allergy. AB - PURPOSE OF REVIEW: Reduced activity of regulatory T cells has recently been described in several diseases, including allergy. This concept of an alteration in the balance between the suppression and activation of harmful T helper type 2 immunity in allergy has important potential implications for strategies to prevent and control disease. RECENT FINDINGS: The past year has seen several important advances in analysing the determinants of this balance and models for inducing tolerance through regulatory T cells, including several different subtypes. These include the recognition that although T helper type 2 responses drive atopic sensitization, the expression of disease involves additional factors, and of a potential role for regulatory T cells in the development of neonatal tolerance. In addition to CD4CD25 regulatory T cells, experimental protocols for the induction of IL-10-producing, transforming growth factor beta and T helper type 1 regulatory T cells have been described in mouse models of airway disease. IL-6 and co-stimulation have been identified as potential determinants of the balance between the suppression and activation of allergic responses. SUMMARY: Different strategies for inducing regulatory T cells in animal models of allergic inflammation and an improved understanding of the factors accentuating or reducing suppression have identified important questions for future translational research. PMID- 16264335 TI - Peptide-based vaccination: where do we stand? AB - PURPOSE OF REVIEW: Allergen-specific immunotherapy represents the only causative approach towards allergy treatment. Specific immunotherapy can, however, include allergic reactions and occasionally life-threatening anaphylaxis. Peptides have been evaluated as a potential therapeutic approach in atopic allergic disease because they have the potential to inhibit T-cell function but not induce anaphylaxis. RECENT FINDINGS: Data from early clinical trials of peptide vaccination revealed that therapy was associated with a modest improvement in allergic disease, and was accompanied by a high frequency of adverse reactions. More recent studies have demonstrated improved clinical outcomes, improved safety, and have defined the mechanisms of adverse events observed in earlier studies. Mechanisms of peptide vaccination include the hyporesponsiveness of allergen-specific responses and the induction of regulatory T cells and cytokines. Novel peptide design has allowed the generation of fragments that contain T-cell stimulatory epitopes, lack B cell epitopes, and can induce protective IgG responses in both mice and humans. Other approaches have focused on hypoallergenic B-cell epitopes that induce inhibitory IgG antibodies. Peptides that specifically induce regulatory cytokine production would also enhance peptide vaccines. Several recent studies have described immunodominant epitopes from major allergens that may form candidate peptides for use in peptide vaccination. SUMMARY: The manipulation of peptide epitopes may provide a strategy for the rational design of peptide allergy vaccines further improving safety and efficacy. PMID- 16264336 TI - Advances in development of hypoallergenic latex immunotherapy. AB - PURPOSE OF REVIEW: The characterization of clinically relevant latex allergens and the production of recombinant allergens is now well advanced, but this knowledge needs to be translated into new strategies for the safe and effective specific treatment of latex allergic diseases including asthma and anaphylaxis. RECENT FINDINGS: The current status of latex allergy is discussed indicating a changing demographic paradigm. A new wave of latex allergy is emerging outside the healthcare setting with the widespread use of latex products. An increased prevalence in developing countries is also reported. Limited studies on current specific immunotherapy for latex allergy are reviewed, confirming the feasibility but demonstrating an unacceptable risk of adverse events. The characterization of latex allergens and the identification of B and T-cell epitopes point to rational strategies for the generation of hypoallergenic preparations for specific immunotherapy. Results to date for latex allergens are reviewed, including recombinant, chemical modification and synthetic peptide approaches. Candidate hypoallergenic preparations for targeting sensitization to the major allergens Hev b 1, Hev b 3, Hev b 5 and Hev b 6.01 have been identified. Further investigations of optimal regimens for the delivery of specific immunotherapy to induce regulatory T-cell function are warranted. SUMMARY: The findings point to the selection of suitable hypoallergenic preparations for clinical trials of effective and safe latex allergy immunotherapy. PMID- 16264337 TI - Role of bacillus Calmette-Guerin as an immunomodulator for the prevention and treatment of allergy and asthma. AB - PURPOSE OF REVIEW: As an essential part of the hygiene hypothesis, the association between exposure to mycobacterial components and the prevention, development and severity of atopic diseases has not been fully understood. The current status on the causal-effect link of this relationship and the potential use of mycobacterial adjuvants as a preventive or disease-modifying modality in allergic diseases is reviewed in this article. RECENT FINDINGS: Data obtained from human and animal models indicate a discrepancy regarding the preventive and therapeutic effect of bacillus Calmette-Guerin in atopic diseases. Among the issues that require clarification include whether the distinction in T helper type 1/2 cells described in mice can be fully extrapolated to humans. Other factors involved could be caused by genetic variation, optimal timing, dose, route of delivery as well as environmental factors, which affect the degree of natural exposure to pathogenic or saprophytic mycobacteria. SUMMARY: Most of the evidence available to date suggests a need for an improved mycobacterial vaccine administered early in life, by means of alternative routes, preferentially mucosal. As switching away from the T helper type 2 immune response by inducing T helper type 1 is unable to explain the underlying mechanisms of action of mycobacterial antigens, it may be worthwhile to investigate whether T regulatory cells are induced in response to different mycobacterial adjuvants. PMID- 16264339 TI - Cross-reactivity of pollen allergens: recommendations for immunotherapy vaccines. AB - PURPOSE OF REVIEW: This review will summarize recent research on pollen allergen and epitope cross-reactivity. Knowledge of these relationships aids in the rational formulation of allergen immunotherapy vaccines. RECENT FINDINGS: There has been further clarification of panallergens and their roles as both major and minor allergens. Recent studies have targeted non-specific lipid transfer proteins and calcium-binding proteins (polcalcins), as well as pathogenesis related protein families and profilins. Polcalcins and non-specific lipid transfer proteins are responsible for pollen-fruit interactions as well as pollen cross-reactivity, in some cases, but not all, accounting for major allergenicity. Delineation of the enzymatic activity of certain allergens explains the ubiquitous nature of these pollen proteins. SUMMARY: Characterization of specific pollen allergens and their protein families has provided insight into the grounds for cross-reactivity. Continuing clarification of these relationships will allow the substitution and consolidation of inhalant extracts as described in the conclusion. PMID- 16264340 TI - Bibliography. Current world literature. Primary immune deficiency disease. PMID- 16264341 TI - Neuro-ophthalmology does have a future. PMID- 16264338 TI - Towards immunotherapy for peanut allergy. AB - PURPOSE OF REVIEW: Food allergy is a major cause of life-threatening hypersensitivity reactions. Peanut allergy is the most serious of the hypersensitivity reactions to foods due to its persistence and high risk of severe anaphylaxis. Currently, strict avoidance of the allergenic food and ready access to self-injectable epinephrine is the 'standard of care' for food allergy. Based on extensive characterization of food allergens and a better understanding of the immunological mechanisms underlying allergic disease, promising therapeutic modalities for food allergy treatment and prevention are being developed. RECENT FINDINGS: Immunotherapeutic strategies include peptide immunotherapy, mutated protein immunotherapy and DNA immunization, which all strive to decrease the deleterious Th2 response. Another approach already in clinical trials for peanut allergy is the anti-IgE therapy which prevents circulating IgE from binding to effector cells, consequently decreasing clinical symptoms after peanut ingestion. In order to be applicable, these strategies must be well tolerated, inexpensive and easily administered. Realistic treatment options would likely involve a combination of different approaches. SUMMARY: Food allergy affects approximately 4-6% of children and 3-4% of adults. Peanut allergy can be devastating as reactions range from urticaria to severe anaphylactic shock and death. The only preventive measure for peanut allergy is strict avoidance of the incriminating food. It is likely immunotherapy will be available in the near future as a well tolerated and effective therapy for treating peanut allergy. The use of the anti-IgE therapy in conjunction with other immunotherapy would possibly be the best treatment option in the future. PMID- 16264342 TI - Neuro-ophthalmic considerations in trigeminal neuralgia and its surgical treatment. AB - PURPOSE OF REVIEW: In this review, we will briefly discuss the clinical manifestations and etiology of trigeminal neuralgia, outline the neuro-ophthalmic features of trigeminal neuralgia, and detail the neuro-ophthalmic side effects and complications of the surgical treatment of trigeminal neuralgia. RECENT FINDINGS: There is a variety of surgical treatment modalities available for patients with trigeminal neuralgia intolerable or resistant to medical therapy. Significant ocular and visual morbidity can result from the surgical treatment of trigeminal neuralgia. SUMMARY: Percutaneous or open surgical procedures for trigeminal neuralgia can result in corneal anesthesia, neurotrophic keratitis, exposure keratitis, herpetic keratitis, ocular motor cranial neuropathies, and optic neuropathy. Ophthalmologists should be aware of these potential problems because they may need to evaluate and provide care to patients with ocular or visual complaints following surgery for trigeminal neuralgia. PMID- 16264343 TI - Nonarteritic anterior ischemic optic neuropathy. AB - PURPOSE OF REVIEW: Nonarteritic anterior ischemic optic neuropathy remains a disease that is poorly understood in many aspects. The clinical presentation may diverge significantly from the classic unilateral, painless, sudden vision loss affecting patients over the age of 50 years. This variability might make nonarteritic anterior ischemic optic neuropathy hard to differentiate from optic neuritis and arteritic ischemic optic neuropathy. The course of nonarteritic anterior ischemic optic neuropathy is also variable, often sequentially affecting the other eye. RECENT FINDINGS: Visual recovery has been reported, but it is not the rule. Multiple risk factors have been proposed, including crowded disc, atherosclerosis, diabetes, hyperlipidemia, hypertension, hypotension, hemoconcentration, hemodilution, and hypercoagulable states. The optic nerve damage in nonarteritic anterior ischemic optic neuropathy appears to result from a perfusion insufficiency in the short posterior ciliary arteries leading to infarction of the retrolaminar portion of the optic disc. The underlying mechanisms are still unclear, however. Multiple medical and surgical treatment options have been investigated, including optic nerve sheath decompression, standard and megadose corticosteroids, levodopa, carbidopa, hyperbaric oxygen, and neuroprotective agents, but no proven effective treatment is currently available. SUMMARY: Intense investigations in humans and animals are under way. Hopefully these studies will enhance our understanding of the risk factors and pathophysiology of nonarteritic anterior ischemic optic neuropathy and aid in developing new strategies for prevention and treatment. PMID- 16264344 TI - Optical coherence tomography: measuring in-vivo axonal survival and neuroprotection in multiple sclerosis and optic neuritis. AB - PURPOSE OF REVIEW: This review considers the latest developments in the use of optical coherence tomography in neuro-ophthalmology. RECENT FINDINGS: Optical coherence tomography can accurately and reproducibly quantitate the micron thickness of the peripapillary retinal nerve fiber layer, as well as the thickness and volume of the macula. It is able to perform both cross-sectional and longitudinal studies in patients with multiple sclerosis and optic neuritis. It is able to measure and assess axonal preservation and protection when used in clinical trials. SUMMARY: Specific guidelines when undertaking optical coherence tomography analyses for patients with multiple sclerosis and optic neuritis are needed to ensure uniformity among clinical trials; this development would be similar to the parameters devised when magnetic resonance imaging emerged as an important technology. PMID- 16264345 TI - Advances in the diagnosis and treatment of ptosis. AB - PURPOSE OF REVIEW: The surgical correction of blepharoptosis, both congenital and acquired, has been intensively examined and reported on for many years. This paper reviews recent publications on basic science, evaluation, technique modifications, and innovative materials in the care of ptosis patients. RECENT FINDINGS: The frontalis suspension technique is a commonly performed surgical correction of congenital blepharoptosis, used widely in the repair of ptosis with poor levator function. The repair typically includes using either tissue such as autologous or banked fascia lata or permanent suture material. The procedure involves connecting the motor unit (frontalis muscle) and the upper eyelid. Authors have recently reintroduced the technique of a dynamic frontalis muscle flap tunneled into the eyelid that directly attaches to the tarsal plate.Patients presenting with symptomatic blepharoptosis due to disinsertion or thinning of the levator aponeurosis require surgical repair. Multiple groups have tended toward a minimally invasive approach directed specifically at the levator aponeurotic defect. Proposed advantages of a small eyelid incision (8-13 mm) include less local anesthetic and tissue distortion, less ecchymosis and edema, decreased operative times, a shortened recovery period, and improved surgical results. SUMMARY: Surgical correction of congenital blepharoptosis may be performed with autologous fascia lata, cadaveric allograft, or permanent suture material. The use of a frontalis muscle advancement flap is elegantly designed; however, its role in clinical practice remains to be defined. Advancement of the levator aponeurosis for senile blepharoptosis may be preformed via a minimally invasive small incision approach. Patients benefit with decreased operative time, edema, ecchymosis, and recovery times. PMID- 16264346 TI - Update on Fuchs' uveitis syndrome. AB - PURPOSE OF REVIEW: This update aims to summarize the current knowledge on Fuchs' uveitis syndrome and review publications in the last 10 years. Theories on the pathogenesis and etiology of Fuchs' uveitis syndrome are revisited and the management of cataract and glaucoma is updated to reflect recent surgical change. RECENT FINDINGS: Retrospective reviews have highlighted that patients with Fuchs' uveitis syndrome are often initially misdiagnosed. Studies comparing local inflammatory mediators and cell types have found differences in Fuchs' uveitis syndrome, but the differences in steroid response and degree of inflammation remain poorly understood. Local production of antibodies to rubella has been recently reported in the aqueous of all patients with Fuchs' uveitis syndrome and no controls. Excellent visual outcomes from phacoemulsification have been reported with reduced complications compared with extracapsular cataract extraction. SUMMARY: Although a single etiological agent and a sensitive laboratory test for the diagnosis of Fuchs' uveitis syndrome is alluring, the diagnosis of Fuchs' uveitis syndrome remains clinical, at least for now. Phacoemulsification has increased the safety of cataract extraction, and the use of intraocular lens is generally safe. The ideal lens material and design are not yet known, but silicone lenses may be best avoided. Glaucoma is often resistant to treatment and should actively be screened for in patients with Fuchs' uveitis syndrome. Medical and surgical treatment for reducing intraocular pressure should be especially aggressive in these patients. Vitrectomy appears to be safe in patients with visually significant vitreous opacification. PMID- 16264347 TI - Presumed ocular histoplasmosis syndrome. AB - PURPOSE OF REVIEW: Presumed ocular histoplasmosis syndrome remains a significant cause of visual morbidity. We review recent literature on the pathogenesis and particularly on the treatment of complications of presumed ocular histoplasmosis syndrome. RECENT FINDINGS: The pathogenesis of presumed ocular histoplasmosis syndrome remains mysterious; although some recent molecular evidence suggests a direct link between Histoplasma capsulatum and presumed ocular histoplasmosis syndrome, other reports document nearly identical disease occurring in the absence of H. capsulatum seropositivity. Treatment options have advanced rapidly in the past few years. Small case series and clinical trials suggest excellent efficacy for photodynamic therapy in the treatment of subretinal neovascularization associated with presumed ocular histoplasmosis syndrome. Preliminary studies also suggest efficacy of intravitreal corticosteroids. A large randomized controlled clinical trial failed to show efficacy for subretinal surgery in the management of presumed ocular histoplasmosis syndrome, however. SUMMARY: Despite continued debate on the etiology of presumed ocular histoplasmosis syndrome, there have been significant advances in treatment of the blinding complications of presumed ocular histoplasmosis syndrome. PMID- 16264348 TI - Bibliography. Current world literature. Neuro-ophthalmology. PMID- 16264349 TI - Myelosuppressive side-effects of radioimmunotherapy of non-Hodgkin's lymphoma: is there an increased risk? PMID- 16264350 TI - Phase I/II clinical trial of the humanized anti-EGF-r monoclonal antibody h-R3 labelled with 99mTc in patients with tumour of epithelial origin. AB - AIM: To evaluate the biodistribution, internal radiation dosimetry and toxicity of the humanized MAb h-R3 labelled with Tc in humans. METHODS: Twenty-five patients with suspected epithelial-derived tumours were included in this study and divided into two groups: group I consisted of 10 patients who received 3 mg/1110 MBq (3 mg/30 mCi); and group II consisted of 15 patients who received 6 mg/2220 MBq (6 mg/60 mCi). Single photon emission computed tomography (SPECT) and planar images, and multiple blood and urine samples were collected up to 24 h after injection. Haematological parameters and adverse effects were classified according to the WHO criteria. Biodistribution, human anti-mouse antibody (HAMA) response and absorbed doses were estimated and reported. RESULTS: Liver, spleen, kidneys and heart were identified as source organs. Their higher uptakes were 53.3+/-6.4%ID, 2.0+/-1.4%ID, 9.8+/-4.3%ID and 2.8+/-0.9%ID, respectively. The urinary bladder and large intestine also had a significant uptake. The mean urinary excretion was around 22%ID. The liver received the highest absorbed doses followed by the kidneys and the urinary bladder wall. There were no haematological or biochemical abnormalities with clinical significance related to the product. No patient developed HAMA response. Preliminary analysis of clinical results showed a sensitivity of 76.5% and a specificity of 100%. CONCLUSIONS: The results of this study suggest that Tc-h-R3 could be used in patients in a safe and effective way, for the diagnosis of epithelial-derived tumours at the two evaluated dose levels. PMID- 16264351 TI - FDG PET in patients with cancer of an unknown primary. AB - BACKGROUND: This prospective study was undertaken to address the capacity of positron emission tomography (PET) with 2-[18F]fluoro-2-deoxy-D-glucose (18F-FDG) to determine the primary tumour site of carcinomas with unknown primary site. PATIENTS AND METHODS: Twenty-five patients with metastases from adenocarcinoma or undifferentiated carcinoma of unknown primary site (CUP) were included prospectively. For all patients, extensive imaging was unsuccessful in localizing the primary site. Patients received 370 MBq of 18F-FDG intravenously, and whole body images were acquired 60 min after injection. All hot spots that could not be attributed to a metastatic site were considered as the primary tumour. The evaluation of FDG PET data was based on clinical and radiological outcome or surgery if indicated. RESULTS: Twenty-four patients were eligible for analysis. All known metastases were visualized. In six patients, FDG PET showed a primary tumour site which was confirmed by follow-up or surgery. In five patients, the primary tumour site was suggested by FDG PET but not confirmed by clinical outcome. No primary tumour was found in the other patients, with a mean follow-up of 15 months. CONCLUSION: In our series, FDG PET allowed the identification of primary tumour site in one quarter of patients with CUP (6/24). We conclude that FDG PET has a place in the initial staging of these patients. PMID- 16264352 TI - Anti-angiogenic therapy and chemotherapy affect 99mTc sestamibi and 99mTc-HL91 accumulation differently in tumour xenografts. AB - BACKGROUND: Favourable effects of cytotoxic chemotherapy for tumours are characterized by the reduced accumulation of radiotracers such as 99mTc sestamibi (MIBI). Anti-angiogenic therapy is primarily cytostatic; consequently, its influence on tracer accumulation may differ from that of cytotoxic treatments. METHODS: Anti-angiogenic therapy employing 2-methoxyestradiol was administered in mice bearing subcutaneous xenografts of LS180 colon cancer cells. The effects of chemotherapy with 5-fluorouracil were examined as a cytotoxic counterpart. Treatments were conducted for 4 days from day 8. Distribution of 99mTc-MIBI and Tc-HL91, a hypoxic marker, was observed on days 8 and 12. Oxygen tension (PO2) in tumours was measured by a microelectrode. Cellular uptake of tracers was examined in vitro in normoxic and hypoxic conditions. RESULTS: 99mTc-MIBI accumulation decreased with increasing tumour weight when no treatment was conducted. Tumour growth was suppressed by anti-angiogenic therapy and chemotherapy. 99mTc-MIBI accumulation in tumours decreased after chemotherapy as compared to pre therapeutic values, whereas accumulation of 99mTc-HL91 increased. In contrast, accumulation of tracers did not significantly change after anti-angiogenic therapy as compared to that observed pre-therapeutically. Tumour PO2 decreased with increasing tumour volume when no treatment was conducted. Chemotherapy reduced PO2 in tumours. PO2 in tumours treated with anti-angiogenic therapy was as high as that observed before treatment. 2-Methoxyestradiol or 5-fluorouracil did not significantly affect tracer accumulation in cells under both normoxic and hypoxic conditions in vitro. CONCLUSION: These findings indicate that scintigraphic assessment of therapeutic efficacy of anti-angiogenic therapy should be performed from a perspective distinct from that of cytotoxic treatment. PMID- 16264353 TI - Role of 99mTc-hexakis-2-methoxyisobutylisonitrile for detecting marrow metastases in childhood solid tumours. AB - AIM: To evaluate the role of 99mTc-hexakis-2-methoxyisobutylisonitrile (99mTc MIBI) for detecting bone marrow metastases in childhood solid tumours, including lymphomas. METHODS: Twenty-six children (18 males, eight females) were studied. They all had proven malignant solid tumours [Hodgkin's lymphoma (5), non Hodgkin's lymphoma (3), neuroblastoma (9), Ewing's sarcoma (3), Langerhans cell histiocytosis (4), rhabdomyosarcoma (1) and germ cell tumour (1)] with suspected bone marrow metastases. All patients underwent computed tomography and/or magnetic resonance imaging, 99mTc-MIBI and Tc-methylene diphosphonate bone scans and bone marrow aspiration and/or biopsy. The scintigraphic evaluation of 99mTc MIBI scans was performed according to the visual assessment of the extent and intensity of uptake. The scintigraphic score, which is the sum of the extent and intensity of uptake, was calculated for each patient. Scores of more than 2 were considered to be positive for bone marrow involvement. RESULTS: Twenty-seven 99mTc-MIBI scans were studied for 26 patients. Twenty-two 99mTc-MIBI scans were accepted as normal bone marrow. Bone scans were also normal in these patients. Five of the 27 99mTc-MIBI scans had scores of more than 2. Bone marrow cytology revealed bone marrow metastases in these patients. CONCLUSION: Abnormal 99mTc MIBI uptake correlated extremely well with bone marrow aspiration/biopsy cytology results. Non-invasive 99mTc-MIBI imaging in children with malignant solid tumours appears to be promising for the evaluation of bone marrow metastases. PMID- 16264354 TI - Use of the gamma probe in sentinel lymph node biopsy in patients with prostate cancer. AB - OBJECTIVE: To describe the reproducibility of the sentinel lymph node technique in patients with prostate cancer and verify if there is improved accuracy over modified lymphadenectomy. MATERIAL AND METHODS: Twenty-three patients with biopsy proven prostate cancer were enrolled in this study. Lymphoscintigraphy was performed after the transrectal administration of Tc sulfur colloid guided by ultrasound, with one injection in each prostate lobe. Images were obtained 15 and 180 min after injection. Sentinel lymph node was harvested during surgery using a gamma probe, followed by extended lymphadenectomy. RESULTS: The mean age of the patients in this study was 66 years. An average of 3.36 sentinel lymph nodes was found for each patient. Radioactive lymph nodes were identified by the gamma probe in 21 out of 23 patients. In one of the patients there was no radiopharmaceutical migration from the injection site and in another the sentinel lymph node was visualized by lymphoscintigraphy but was not found during surgery. Three patients had lymph node metastasis; in one of these patients the sentinel lymph node was the only positive node and was found outside the modified lymphadenectomy region (obturator fossa and the external iliac). CONCLUSION: Sentinel lymph node biopsy in prostate cancer adds important information to the staging of patients, not always attained through the lymphadenectomy restricted to the obturator fossa and external iliac. Such information is essential for the choice of the best treatment to be applied. PMID- 16264355 TI - Scintigraphic sentinel node detection in breast cancer patients: paired and blinded comparison of 99mTc dextran 500 and 99mTc phytate. AB - BACKGROUND: Breast cancer surgery has evolved towards minimizing morbidity, maximizing cure rates and stratifying treatments according to disease stage. Sentinel lymph node biopsy is becoming standard practice in most centres. However, no standard radiopharmaceutical exists. OBJECTIVES: To blindly compare 99mTc dextran 500 and 99mTc phytate in the scintigraphic detection of sentinel lymph nodes. Endpoints were the detection of axillary or internal mammary lymph nodes, number of lymph nodes detected, detectability in the first versus the second hour of evaluation and the relationship between rate of detection and age or body mass index. METHODS: Forty-six patients with histological diagnosis of early breast cancer, without previous surgical treatment, were enrolled in our study. Each patient underwent lymphoscintigraphy twice: on one day with 99mTc dextran 500 and on another day with 99mTc phytate. Images were acquired 1 h and 2 h after tracer administration. RESULTS: Eighty-eight lymphoscintigraphic studies were performed in 44 patients. On the first image (taken at 1 h), 34 patients from the 99mTc dextran group showed sentinel nodes compared with 28 positive examinations using 99mTc phytate (P = 0.113). On the second image (taken at 2 h) 39 patients from the 99mTc dextran group showed positive results compared to 30 positive examinations using 99mTc phytate (P = 0.036). There was no statistically significant correlation between the body mass index or age and the result of the lymphoscintigraphy. CONCLUSION: 99mTc dextran 500 is better than 99mTc phytate for use in a 2 h interval scintigraphic protocol because it demonstrates the sentinel node in a significantly higher number of patients and also showed more lymph nodes suitable for pathological examination. PMID- 16264356 TI - Increasing the radiochemical purity of 99mTc sestamibi commercial preparations results in improved sensitivity of dual-phase planar parathyroid scintigraphy. AB - BACKGROUND: Poor results for dual-phase parathyroid scintigraphy have recently prompted increased use of dual-tracer imaging. We noticed that seminal studies used higher radiochemical purity than provided by current commercial preparations meeting US Pharmacopea (USP) specifications (90% of technetium bound to sestamibi). We surmised that the presence of unbound Tc (non-MIBI tracer) might hamper dual-phase detection that is dependent on rapid wash-out of technetium from thyroid tissue. PURPOSE: To test the hypothesis that reducing non-MIBI tracer will enhance thyroid wash-out and improve sensitivity of dual-phase imaging. METHODS: Starting in April 2003 we decreased the technetium to sestamibi ratio. This resulted in a significant decrease of non-MIBI tracer from 8.1+/-2.2% (SD) (group 1, n = 42) to 3.5+/-1.1% (group 2, n = 47) (P < 0.05 t-test). We performed a retrospective review of 89 patients with primary hyperparathyroidism who underwent imaging and subsequent surgery. The pathological findings served as the 'gold standard'. RESULTS: Scanning detected 21/39 diseased glands (sensitivity=54%) in group 1 patients. In group 2 imaging detected 38/45 diseased glands (sensitivity = 84%). An improvement in sensitivity (P < 0.01) was achieved by modifying the radiopharmaceutical preparation. CONCLUSIONS: Elevated levels of non-MIBI tracer in Tc-MIBI commercial preparations result in persistent thyroid background activity that may interfere with detection of parathyroid pathology. Achieving a higher degree of radiochemical purity (at least 95% bound, 5% impurities) than required by USP may be needed for optimal results. The large variation in sensitivity reported in the literature may be related in part to non uniform radiopharmaceutical preparation. PMID- 16264358 TI - FDG PET imaging in the evaluation of post-transplant lymphoproliferative disorder following renal transplantation. AB - OBJECTIVES: To assess the usefulness of fluorodeoxyglucose positron emission tomography (FDG PET) in the evaluation of post-transplant lymphoproliferative disorder (PTLD) following renal transplantation. METHODS: Staging FDG PET examinations were reviewed in five patients with biopsy proven post-renal allograft PTLD and compared with conventional imaging. Follow-up PET examinations of three patients at various points during their therapy for PTLD were also reviewed and compared with clinical outcome. RESULTS: PET imaging identified all disease sites seen at cross-sectional imaging, clarified sites of uncertain significance and detected additional disease in two out of five patients. Early follow-up imaging impacted PTLD management in two of three patients. CONCLUSION: In this limited series FDG PET was an effective imaging modality in the staging and follow-up of PTLD. PMID- 16264357 TI - The contribution of statistical parametric mapping in the assessment of precuneal and medial temporal lobe perfusion by 99mTc-HMPAO SPECT in mild Alzheimer's and Lewy body dementia. AB - AIM: To assess the role of 99mTc-hexamethylpropyleneamine oxime single-photon emission computed tomography (99mTc-HMPAO SPECT) imaging of the precuneus and medial temporal lobe in the individual patient with mild Alzheimer's disease and dementia with Lewy bodies (DLB) using statistical parametric mapping and visual image interpretation. METHODS: Thirty-four patients with mild late-onset Alzheimer's disease, 20 patients with early-onset Alzheimer's disease, 15 patients with DLB and 31 healthy controls were studied. All patients fulfilled appropriate clinical criteria; the DLB patients also had evidence of dopaminergic presynaptic terminal loss on 123I-N-omega-fluoropropyl-2beta-carbomethoxy-3beta (4-iodophenyl)-tropane imaging. 99mTc-HMPAO SPECT brain scans were acquired on a multidetector gamma camera and images were assessed separately by visual interpretation and with SPM99. RESULTS: Statistical parametric maps were significantly more accurate than visual image interpretation in all disease categories. In patients with mild late-onset Alzheimer's disease, statistical parametric mapping demonstrated significant hypoperfusion to the precuneus in 59% and to the medial temporal lobe in 53%. Seventy-six per cent of these patients had a defect in either location. No controls had precuneal or medial temporal lobe hypoperfusion (specificity, 100%). Statistical parametric mapping also demonstrated 73% of patients with DLB to have precuneal abnormalities, but only 6% had medial temporal lobe involvement. CONCLUSION: These findings illustrate the capability of statistical parametric mapping to demonstrate reliable abnormalities in the majority, but not all, patients with either mild Alzheimer's disease or DLB. Precuneal hypoperfusion is not specific to Alzheimer's disease and is equally likely to be found in DLB. In this study, medial temporal hypoperfusion was significantly more common in Alzheimer's disease than in DLB. Statistical parametric maps appear to be considerably more reliable than simple visual interpretation of 99mTc-HMPAO images for these regions. PMID- 16264359 TI - High prevalence of oesophageal involvement in patients with undifferentiated connective tissue disease using radionuclide oesophageal transit scintigraphy. AB - AIM: To look for the frequency of oesophageal dysfunction using radionuclide oesophageal transit scintigraphy in 145 patients with undifferentiated connective tissue disease (UCTD); to seek the correlation between the clinical/laboratory data and scintigraphic alterations; and to determine predictive value of radionuclide oesophageal transit scintigraphy for evolution to established connective tissue disease (CTD). METHOD: One hundred and forty-five patients with UCTD were examined by 99mTc-DTPA oesophageal transit scintigraphy. The intraoesophageal transport of the radiopharmaceutical was followed and imaged by a gamma camera, a series of 128 x 128 images were stored and evaluated. The correlation between the scintigraphic data and clinical and laboratory parameters was analysed statistically. RESULTS: Unequivocally positive scintigraphy, indicative of motor abnormality was found in 46% of patients (66), 71% (47) of whom were totally asymptomatic. Significant correlation was found between the presence and severity of scintigraphic alterations and antinuclear antibodies, the anti-beta2GPI, IgM, IgG, the aCL antibody positivity, and the skin symptoms. Scintigraphic positivity was significantly more frequent in patients evolving to definitive CTD (P = 0.0178), and abnormal scan predisposed to transition into the definitive CTD (odds ratio, 2.292; CI, 1.610-4.525). Its cumulative positive predictive value was found to be 43% and cumulative negative predictive value 73% with regard to the development of a definitive CTD. CONCLUSION: Our results show that scintigraphic alterations together with clinical and laboratory alterations can help the clinician in the prediction of final outcome. PMID- 16264360 TI - 99mTc-HL91 is inferior to 201Tl in scintigraphic detection of chronic myocardial ischaemia. AB - AIM: 99mTc-HL91 is a new hypoxia agent and can identify acutely ischaemic viable myocardium in a canine model using a standard gamma camera. The purpose of the study was to determine whether this tracer could be used to detect regional ischaemia in chronic ischaemic myocardium in a swine model. METHOD: Using a porcine model of chronic myocardial ischaemia, five mini-pigs with proximal left anterior descending artery (LAD) stenoses were studied. Injection of 462.5 MBq (12.5 mCi) 99mTc-HL91 was followed by imaging over 2 h. Coronary angiography and dipyridamole stress-re-injection 201Tl single photon emission computed tomography (SPECT) were performed within 1 day of each other. RESULTS: None of the five pigs demonstrated positive hot 99mTc-HL91 uptake throughout the 2 h imaging, whereas four of the five animals showed significant myocardial ischaemia on 201Tl SPECT. The region of interest analysis of LAD/left circumflex artery count ratios at 1 and 2 h demonstrated similar 99mTc-HL91 uptake and retention in chronic ischaemic as well as non-ischaemic myocardium. CONCLUSION: 99mTc-HL91 is inferior to 201Tl in scintigraphic detection of chronic myocardial ischaemia. PMID- 16264361 TI - Interdependence between measures of extent and severity of myocardial perfusion defects provided by automatic quantification programs. AB - AIM: To evaluate the accuracy of the values of lesion extent and severity provided by the two automatic quantification programs AutoQUANT and 4D-MSPECT using myocardial perfusion images generated by Monte Carlo simulation of a digital phantom. The combination between a realistic computer phantom and an accurate scintillation camera simulation tool allows the generation of realistic single-photon emission computed tomography (SPECT) images similar to those obtained in clinical patient studies. METHODS: The NCAT phantom and the SIMIND Monte Carlo program were used to simulate myocardial perfusion studies. Perfusion defects with sizes ranging from 5 to 17% of the left ventricular wall volume and reductions in tracer uptake of 20, 60 and 100% were simulated in three vascular territories. RESULTS: The values of the extent provided by the programs were dependent on the reduction in tracer uptake, i.e. the severity. Similarly, the measures of severity were dependent on the size of the lesions. The severity provided by AutoQUANT for different defects was not dependent on the location, whereas 4D-MSPECT presented different values depending on the location in the left ventricle. The measures of extent and severity of the defects with the same true extent and activity uptake reduction provided by the two programs were different. CONCLUSIONS: The NCAT phantom and the SIMIND Monte Carlo program were shown to be useful in simulating clinical myocardial SPECT studies. The quantification programs gave values of lesion extent that were dependent on the magnitude of the severity. Users should therefore consider this dependence when interpreting results from these programs. PMID- 16264362 TI - Variation of DaTSCAN quantification between different gamma camera types. AB - OBJECTIVE: To acquire data from a 123I filled Alderson phantom on different gamma cameras types and compare the relative uptake results from processing using the QuantiSPECT program (GE Healthcare). METHODS: A DaTSCAN phantom was filled using the standard protocol and imaged on seven different gamma camera types and on two identical cameras of the same type. The standard GE Healthcare protocols for the given cameras were used. Aliquots of the striatum and brain background were counted in a gamma counter to determine variations in filling concentration. All the raw DaTSCAN SPECT data was imported into QuantiSPECT and processed by the three different algorithms (two box, three box and crescent) to determine the relative uptake in the striatum. Inter-operater and intra-operator variation was also determined. RESULTS: The 10% variation in filling concentration found across the sites was compensated for in the final results. There was a 5-15% variation between cameras depending on the processing algorithm used. There was an intra operator variation of between 5 and 12% which reflected the proportion of operator intervention within the processing method. There was no statistical variation between operators. CONCLUSIONS: The transfer of a DaTSCAN database between camera types is feasible, but ideally all data would be acquired on a single camera type and phantom data used to normalize the database accordingly. PMID- 16264363 TI - Comparison of different methods of DatSCAN quantification. AB - BACKGROUND: The quantification of DaTSCAN images can be used as an adjunct to visual assessment to differentiate between Parkinson's syndrome and essential tremor. Many programs have been written to assess the relative uptake in the striatum. AIM: To compare two of the commercially available programs: QuantiSPECT, which analyses isolated data in two dimensions, and BRASS, which performs three-dimensional processing referencing a normal image template. METHOD: Twenty-two patients (11 with Parkinson's syndrome and 11 with essential tremor) were visually assessed by two nuclear medicine consultants. The patient data were then processed using two commercial programs to determine the relative uptake in the striatum. A comparison of the results from the programs was performed, together with a comparison with the visual assessment. The inter operator and intra-operator variabilities were also ascertained. RESULTS: All programs and processing methods could distinguish between Parkinson's syndrome and essential tremor. There was also a good correlation between the results from the three- and two-dimensional methods. The intra-operator and inter-operator variabilities were dependent on the amount of operator intervention. CONCLUSION: Both programs allowed statistical differentiation between Parkinson's syndrome and essential tremor. Strict operator protocols are needed with QuantiSPECT to reduce inter- and intra-operator variation. The three-dimensional method (BRASS) gave greater concordance than the two-dimensional method (QuantiSPECT) with the visual assessment, but at a cost of increased operator time. PMID- 16264364 TI - Development of a combined audiovisual and extremity dose monitoring software tool for use in nuclear medicine. AB - OBJECTIVES: Conventional extremity dose monitoring in nuclear medicine, using thermoluminescent dosimeters, provides a convenient method of determining integral doses from a series of procedures. Although semiconductor extremity probes are able to add time information and allow doses from individual procedures to be determined, it can be difficult to relate individual operations to the dose-time curve. Solutions to this problem have been identified and developed. METHODS: A novel software tool (Extremity Dose Information Package, EDIP) has been developed that uniquely combines and synchronizes two audiovisual and extremity probe data-streams. The value of this extra information was assessed by acquiring audiovisual and extremity dose information in nuclear medicine and radiopharmacy settings. RESULTS: The ability of the software tool to synchronize audiovisual and dose data-streams was verified. Preliminary studies of handling techniques in radiopharmacy and radioiodine administrations using this tool showed areas in which techniques could be adapted to reduce extremity doses, which would have been difficult or impossible to identify using the dose time information alone. CONCLUSIONS: This low-cost multimedia extremity dose monitoring package can be used, for example, to aid staff training and pinpoint issues with current operating procedures within a clinical nuclear medicine department. Its unique ability to combine and synchronize audiovisual and dosimetry data is also likely to be of benefit to other industries handling unsealed radioactive materials. PMID- 16264367 TI - Use of 99mTc labelled human immunoglobulin G for sentinel node imaging. PMID- 16264365 TI - Altered gene expression profiles by sodium/iodide symporter gene transfection in a human anaplastic thyroid carcinoma cell line using a radioactive complementary DNA microarray. AB - BACKGROUND: The sodium/iodide symporter (NIS) is a membrane glycoprotein that mediates active 131I uptake during the treatment of cancer of the thyroid gland and extrathyroidal tissues. NIS gene transfection, a gene-therapy modality, has been introduced in many types of cancer, such as prostate cancer and breast cancer, and has demonstrated a high potential for the treatment of non-thyroidal cancers. AIM: To investigate the pattern of NIS gene expression and provide evidence of its beneficial effects in human anaplastic cancer ARO cells by using a radioactive complementary DNA (cDNA) microarray. METHODS: For cDNA microarray data analysis, superimposed images and clustergrams were prepared from basic radioactivity data obtained using a phosphoimager system. Gene expression profiles were constructed using the Z-transformed values of genes related to cancer biology. RESULTS: Radioactive cDNA microarray studies showed that 11 genes were upregulated (Z ratio > 1.5) and 31 genes were downregulated (Z ratio < -1.5) in response to NIS gene transfection. Of these differentially expressed genes, 33% were related to cell proliferation and apoptosis. Moreover, NIS gene transfection into an anaplastic thyroid cancer cell line affected the expression of the protein tyrosine phosphatase (PTP) family and Ras oncogene family, including Ras, Rac and Rab. CONCLUSION: The identification of changes in the patterns of gene expression may provide a better understanding of the response of molecular mechanisms to NIS gene transfection. PMID- 16264371 TI - Influenza vaccine 2005-2006. PMID- 16264372 TI - Montelukast (Singulair) for perennial allergic rhinitis. PMID- 16264374 TI - Morbidity of third molar extraction in patients between 12 and 18 years of age. AB - OBJECTIVE: An analysis is made of the incidence of complications following third molar surgical extraction in patients between 12 and 18 years of age. PATIENTS AND METHOD: A retrospective study was conducted of 390 surgical extractions of upper and lower third molars in 173 patients operated upon under locoregional anesthesia during the year 2000 in the Master in Oral Surgery and Implantology of Barcelona University Dental School (Spain). The patients were divided into three age groups (A: 12-14 years, B: 15-16 years, C: 17-18 years). The reason for extraction, the degree of dental development, and third molar position, angle and impaction were recorded. Finally, the association of these variables to the appearance of postoperative complications was analyzed. RESULTS: Most patients were females (66.9%), and the age group in which most extractions were carried out (62.8%) corresponded to Group C (17-18 years). The main reason for extraction was orthodontist indication (40.5%), closely followed by prophylaxis (39.5%). The existence of clinical manifestations was an indication for extraction in 20% of cases. The postoperative complications rate after the extraction of the global 390 molars was 15.6%. In Groups A, B and C the complication risks were 17.4%, 19% and 13.7%, respectively. All complications were reversible and of short duration. One case each of inferior alveolar nerve paresthesia and lingual nerve paresthesia was recorded, which subsided after one and two months, respectively. CONCLUSIONS: There were no significant differences in complications between the three age groups. An increased tendency towards complications was observed in females, and the percentage of postoperative problems increased. PMID- 16264375 TI - Hereditary familial polyposis and Gardner's syndrome: contribution of the odonto stomatology examination in its diagnosis and a case description. AB - Familial Adenomatous Polyposis (FAP) and its phenotype variant, Gardner's syndrome, constitute a rare autosomal dominant inherited disorder. They are characterised by the development, generally during the second and third decades of life, of multiple adenomatous polyps in the colon and rectum. These polyps have a high risk of subsequently becoming malignant, which normally occurs in the third and fourth decades of life. The phenotypical features of FAP can be very variable. As well as colorectal polyps, these individuals can present with extra colonic symptoms, among which are particularly: gastro-duodenal polyps, dermoid and epidermoid cysts, desmoid tumours, congenital hypertrophy of the retinal pigment epithelium, disorders of the maxillary and skeletal bones and dental anomalies. In this paper the most important aspects of this syndrome are reviewed, showing an example based on a well documented clinical case. The importance of odonto-stomatological examinations should be pointed out, among others, as a means of reaching a presumptive diagnosis, whose confirmation is vital to the patient. PMID- 16264376 TI - Treatment of pemphigus and pemphigoids. PMID- 16264377 TI - Measurement of secondary mucositis to oncohematologic treatment by means of different scale. Review. AB - Oral mucositis is the inflammation that takes place in the oral epithelium, as a result of antineoplastic treatments such as radiotherapy, chemotherapy or bone marrow transplant, being very frequent in these treatments for oncohematologic disease. The consequences of this inflammation, not only affect the quality of life of the patient, but can also suppose a limitation in the application of the treatment, as well as an increase in the hospital stay and therapeutic costs. A main obstacle for the study of the mucositis, has been the lack of a system adapted for its valuation by means of the oral examination. Methods developed to measure and quantify the changes produced in oral epithelium as a result of treatment of cancer can be very varied from more simple methods, such as general scales with four or five degrees of severity that link the mucositis to the state of oral health, to specific scales of treatment. In this last type of scale the type of antineoplastic treatment that gave rise to the mucositis is identified giving a global severity score for the mucositis. The establishment of a common scale for the evaluation of mucositis is important, not only for clinical purposes but also for the investigation of the degree of toxicity of the different therapeutic regimes that give rise to the mucositis. PMID- 16264378 TI - Intraoral tumor with rapid growing. Report of a case. AB - The appearance of an intraoral mass is common in our specialty. Most are benign lesions, but some are primary malignancies. Metastases account for less than 1% of all oral malignancies. An 86 year old woman was referred to our department with a large, asymptomatic, intraoral, fast-growing mass. She had no previous cancer history or other relevant physical findings. The radiology studies showed underlying bone erosion. The histological study showed a metastatic adenocarcinoma with a suspected origin in the abdomen. We were unable to identify it by non invasive diagnostic procedures. Given the patient's general status and despite the ominous prognosis of such lesions, we decided not to perform any aggressive therapy beyond removing the oral mass, in order to maintain her quality of life. There have been no local recurrences until this time. PMID- 16264379 TI - Bone grafting simultaneous to implant placement. Presentation of a case. AB - Bone defects at mandibular alveolar crest level complicate the placement of dental implants in the ideal location. Surgical reconstruction using autologous bone grafts allows implant fixation in an esthetic and functional manner. We describe a patient with large mandibular bone loss secondary to periodontal inflammatory processes. Reconstruction of the mandibular alveolar process was carried out using onlay block bone grafts harvested from the mandible. The grafts were stabilized by positioning the dental implants through them--a procedure that moreover afforded good primary implant fixation. After two years of follow-up the clinical and radiological outcome is good. In the lower jaw, where bone regeneration is complicated, we were able to achieve good results in this patient -minimizing the corresponding waiting time by grafting and placing the implants in the same surgical step. PMID- 16264380 TI - Rescue surgery (surgical repositioning) of impacted lower second molars. AB - The impaction of lower second molars, given that its incidence is 0.03 to 0.21%, is a rare complication in tooth eruption. It has been detected more often in unilateral form than bilateral and is more common in the mandible than in the maxillary. It has a slight predilection for males, and mesial inclination is more usual. A wide variety of therapeutic approaches have been published, basically referring to surgical techniques, independent or complemented by means of orthodontic technical aids, with the aim of placing the tooth in the correct position, and which are encompassed under the concept of surgical rescue. In cases resolved with repositioning of an impacted tooth, prophylactic root extraction has been proposed as obligatory. We present a case of a 12 and a half year old patient referred to the University of Seville due to non-eruption of the left lower second molar. The patient was referred by her orthodontist, who detected the impaction before starting orthodontic treatment. The orthodontist requested that, if it was possible, we did not extract the root of the third molar, because its eruption would be feasible in the future (there would be sufficient space in the arch). The spaces available were measured and we decided to attempt the repositioning of the impacted tooth without extracting the root of the wisdom tooth, which was carried out successfully. PMID- 16264381 TI - Epidermoid carcinoma of the lip: an immunohistochemical study. AB - OBJECTIVES: To determine the expression of the c-erb-B2, p53, bcl-2, Ki67 and CD44varV6 proteins, and to establish their prognostic value in epidermoid carcinoma of the lip. STUDY DESIGN: Immunohistochemical study of the c-erb-B2, p53, bcl-2, Ki67 and CD44varV6 proteins in 79 epidermoid carcinomas of the lip, diagnosed and treated over a period of 20 years. The data obtained were subjected to uni- and multi-variate statistical analyses. RESULTS: Immunostaining was positive in 75% of cases for c-erb-B2 protein, in 70.6% for p53 protein, in 3.8% for bcl-2 protein and in 89.9% for adhesion molecule CD44varV6. Ki67 protein expression varied between a minimum of 0% and a maximum of 6.29%. Most immunohistochemical factors analyzed presented no prognostic value for epidermoid carcinoma of the lip. Only those patients affected by this type of tumor that expressed the adhesion molecule CD44varV6 were significantly associated with a greater survival calculated by means of Kaplan-Meier analysis. CONCLUSIONS: The immunohistochemical techniques analyzed for the anatomicopathological study of epidermoid carcinoma of the lip should not routinely be used due to their high cost and low utility in daily clinical practice. PMID- 16264382 TI - Oral squamous cell carcinoma. Cytometric parameters of prognostic interest. AB - OBJECTIVES: The present study was made in order to find possible prognostic factors in oral squamous cell carcinoma, given that it is a frequent disease (3 4% of all malignant tumors) and is the cause of a high morbidity and mortality which justifies any attempt to contribute something towards the understanding of this pathology. STUDY DESIGN: 81 oral squamous cell carcinomas, treated with the same procedure, and retrieved from the archive of the Hospital Universitario Marques de Valdecilla (Santander) were studied. Flow cytometry was carried out on 67 of the samples. RESULTS: No statistically significant differences were found between the cellular proliferative index and the mitotic index, ploidy and the S phase factor. Likewise, none of the cytometric variables studied presented any association with the appearance of local relapse, distant metastases or survival. CONCLUSIONS: These variables cannot be used as a prognostic factors in squamous cell carcinomas of the oral cavity. PMID- 16264383 TI - Differential diagnosis of temporomandibular joint disorders (TMD). PMID- 16264384 TI - Differential diagnosis of exophytic lesions of soft oral tissue. PMID- 16264385 TI - Epidemiology of the most common oral mucosal diseases in children. AB - Dentists who treat children must be alert to the possibility of finding diseases of the oral mucosa, especially in younger children. The present study aimed to review the most updated information and the experience of our group in order to yield epidemiological data that assist diagnosis of the most common diseases of the oral mucosa in children. Recent epidemiologic studies have shown a wide variability in the prevalence of oral mucosal lesions in different regions of the world and have led researchers to draw disparate conclusions. Moreover, studies have not been designed using standard criteria, further explaining the wide variability in the percentage of different groups of children with oral lesions, which ranges from 4.1 to 52.6%. The lesions most frequently considered by authors and that most often appear in the different studies are: recurrent aphthous stomatitis (0.9-10.8%), labial herpes (0.78-5.2%), fissured tongue (1.49-23%), geographic tongue (0.60-9.8%), oral candidiasis (0.01-37%) and traumatic injury (0.09%-22.15%). Dentists must be able to detect any of the numerous possible disorders and perform the correct differential diagnosis, key to the treatment plan. The aim of this paper, based on a review of the different national and international studies, is to contribute data on the most important oral mucosal diseases in the paediatric population in terms of prevalence and differential diagnosis. PMID- 16264386 TI - Burning mouth syndrome: a retrospective study of 140 cases in a sample of Catalan population. AB - The results of analyzing etiologic and clinical factors, and their connection with the burning mouth syndrome (BMS) in a sample of Catalan (Barcelona, Spain) population are presented in this work. The purpose of this study is to establish connections between BMS and the following variables: age, sex, overt depression, masked depression, cancerophobia, dry mouth, foreign body sensation in the mouth, and burning. 140 clinical cases of patients diagnosed with the disease and 140 cases of control patients are studied here. The data were statistically analyzed to study connections as well as the disease and variables frequency. The obtained results will help understanding possible connections of the studied etiologic and clinical factors with the disease, as well as the course of BMS, and its consequences in the Catalan population. PMID- 16264387 TI - Multifocal epithelial hyperplasia. Report of nine cases. AB - Multifocal epithelial hyperplasia (MEH) is also known as focal epithelial hyperplasia, Heck's disease or multifocal papillomavirus-induced epithelial hyperplasia. It is characterised by the presence of multiple lesions in the oral mucosa of children and it has been associated with the presence of the human papillomavirus. The aim of this study was to determine the clinico-pathological features of the cases diagnosed as MEH in the Service of Dermatology of the Hospital Manuel Gea Gonzalez (SDHMGG). The files of the SDHMGG were reviewed and all cases diagnosed as MEH were retrieved. Nine MEH cases were found. Most of the patients were 20 year-old or younger (67%) and females were more commonly affected (78%). All patients presented multiple lesions and always, close relatives with similar lesions were found. Lesions were located most commonly in the buccal mucosa, lower lip and commissures. MEH is a soft tissue intraoral condition that needs treatment solely of the traumatised lesions or those with cosmetic problems. Remaining lesions will disappear with the age of the patients. It is suggested that this entity should be named multifocal epithelial hyperplasia since this name describes better the clinico-pathological and microscopic features of the disease. PMID- 16264389 TI - The role of bombesin in the mechanism of pituitary hormones release. AB - OBJECTIVES: Functional studies indicate that bombesin may be involved in many physiological functions, including sensory transmission, the regulation of central autonomic pathways, thermoregulation, secretion of pituitary hormones, gastric and pancreatic secretion, food intake and satiety. MATERIAL AND METHODS: In order to evaluate the role of bombesin in the mechanism of pituitary hormones release the effects of bombesin on rGH, rTSH, rPRL, rFSH and rLH release were investigated in female Wistar-Kyoto rats. In studies 'in vitro' bombesin in a dose of 1nM, 10nM, 100nM was administered to pituitary cell culture. After 60, 120, 240 min. of incubation pituitary hormones were measured. In studies 'in vivo' bombesin was injected intraventricularly (icv) in a dose of 0.5 microg/5 microl aCSF (artificial cerebrospinal fluid) for 5 min. (experiment I). In experiment II bombesin was administered intravenously (iv) in a dose of 10 microg. After 60 min, 120 mins the animals were decapitated and serum rGH, rPRL, rTSH, rLH, rFSH concentrations were measured with RIA methods. RESULTS: Bombesin stimulated rLH, rPRL, rTSH release from cultured pituitary cells. A slight increase of GH was also observed. After intravenous (iv) injection of bombesin an increase in serum rPRL and rGH levels was found. However, the intraventricular (icv) administration of bombesin leads to decrease of serum rGH, rPRL and rTSH concentrations. CONCLUSION: Our studies 'in vitro' and 'in vivo' indicate that bombesin may be involved in the modulation of pituitary hormones release. PMID- 16264388 TI - Analgesic efficacy of diclofenac versus methylprednisolone in the control of postoperative pain after surgical removal of lower third molars. AB - OBJECTIVE: To compare the analgesic efficacy of methylprednisolone (corticoid) versus diclofenac (nonsteroidal antiinflammatory-NSAID-) after surgical removal of lower third molars. STUDY DESIGN: Prospective study on 73 patients for the surgical removal of their lower third molars. These patients were separated in two groups at random: a diclofenac group and a methylprednisolone group. A record card was filled in with preoperative and postoperative epidemiological and clinic data. The pain level assessment was made on a semiquantitative and an analogical visual scales and in relation to the amount of rescue analgesics consumed. Pain levels were measured at 1, 8, 24, 48 and 72 hours. In order to make a broad study of data, BMDP program was used for statistics. RESULTS: In the pain described by patients in the analogical visual scale there were no differences between groups as a whole. There were no differences in the amount of rescue analgesics consumed. CONCLUSIONS: There is less pain in the corticoid group but not as to justify its routine use. PMID- 16264390 TI - The role of leptin in the regulation of pituitary hormones release. AB - OBJECTIVE: It has been reported that leptin plays an important role in the regulation of food intake and energy expenditure. There are controversial opinions about effects of leptin on the hormonal system. The aim of this study was to estimate the effect of leptin on pituitary hormones release after central and peripheral administration. METHODS: Leptin was injected intracerebroventricularly (icv) in a dose of 0.5 microg/5microl aCSF (artificial cerebrospinal fluid) for 5 min in Wistar Kyoto rats. At 60 and 120 min after injections the animals were decapitated. Leptin in a dose of 10 microg in 300 microl of saline was administered intravenously (i.v). At 60 mins the animals were decapitated. Serum rLH, rFSH, rPRL, rTSH, rGH concentrations were measured with RIA methods. RESULTS: After central (icv) injection of leptin we observed an increase of rGH, rTSH and a decrease of rPRL. However, after peripheral (iv) injection of leptin we found a decrease of rGH and rTSH and an increase of rPRL. We did not find any significant changes in LH and FSH after icv and iv injection of leptin. CONCLUSIONS: The opposite effects of leptin on pituitary hormones release were observed due to the method of leptin administration. Leptin may play a modulating role in the mechanism of pituitary hormones release. PMID- 16264391 TI - Action of prolactin, prolactin-releasing peptide and orexins on hypothalamic neurons of adult, early postnatally overfed rats. AB - OBJECTIVES: Hypothalamic neurons of rats overweight due to early postnatal overfeeding (SL) differ from those of control rats in their responses to feeding relevant hormones like leptin or insulin. The question arose whether prolactin and prolactin-releasing peptide (PrRP) express also differential action in SL rats. These peptides are described to have an effect on food intake and body weight regulation. Prolactin is co-synthesized in lateral hypothalamic neurons together with orexins that were also analyzed in this study. METHODS: Single unit activity was extracellularly recorded in brain slices from adult control rats (CL) and from rats previously raised in small litters (SL). The action of the peptides on the firing rates was evaluated in the medial parvicellular part of the paraventricular nucleus (PaMP) and the medial arcuate nucleus (ArcM). RESULTS: In control rats, PrRP significantly activated PaMP neurons, whereas prolactin and orexin-A induced also inhibition. In SL rats, there was a significantly different effect of orexin-B on PaMP neurons: the main effect changed from activation in controls to inhibition. ArcM neurons of both control and SL rats were mainly excited by prolactin and orexins. CONCLUSION: Changes acquired during early development of neuronal responses to feeding relevant peptides are not a general non-specific mechanism of neurochemical plasticity, but concern specific hypothalamic nuclei and/ or hormones and neuropeptides. The increase in inhibition by orexin-B of hypothalamic paraventricular neurons could in vivo contribute to the neonatally acquired disposition towards persistingly increased food intake and reduced energy expenditure of overweight SL rats. PMID- 16264392 TI - Effect of regular exercise on health and disease. AB - It is known for a long time that exercise increases physical adequacy, has beneficial effects on the general health condition as well as a playing preventing role against various disease states. To decrease the risk of disease and maintain good health, the natural defense system of the organism needs to be strengthened. It is thought that in addition to increasing the body's resistance to disease through the strengthening of the immune system, decreases the convalescence time, increases work efficiency and improves the sportive performance of the individual all which would contribute positively to the national economy. The positive effects of regular exercising of aerobic nature such as strengthening of the immune system, protection against diseases as well as its positive effects on quality of life will help to emphasize the importance of physical exercise and improve the general view of sports by society. PMID- 16264393 TI - Aluminum-induced neurotoxicity and oxidative damage in rabbits: protective effect of melatonin. AB - OBJECTIVE: The present study was aimed to investigate: (1) the neurotoxic oxidative damage of orally administered aluminum chloride (AlCl3) in rabbits (Biochemical and morphopathological studies). (2) The effect of melatonin as an antioxidant and free radical scavenger on oxidative neuropathic changes. METHODS: Thirty-five male rabbits were divided into 4 groups (A, B, C [10 animals each] and D [5 animals]). Group A received AlCl3 (20 mg/l via drinking water for 3 months). Group B received AlCl3 for 3 months then administered with melatonin (10 mg/kg b.w. sc daily for 15 days). Group C received AlCl3 plus melatonin for 3 months. Group D received the solvent and served as control. Malondialdehyde (MDA) and 4-hydroxyalkenals (4-HDA) as lipid peroxides as well as superoxide dismutase (SOD) as an antioxidant enzyme were measured. Aluminum residue in the brain tissue was measured spectrophotometerically. The morphopathological changes were also examined by light and electron microscopes. RESULTS: MDA and 4-HAD were significantly increased in group A versus those of controls while significantly decreased in groups B and C compared with those of A group. SOD run in an opposite manner. Aluminum concentration was significantly increased in groups A, B and C when compared with group D while it significantly decreased in groups B and C when compared with that of group A. The neuropathlogical examination in the animals of group A revealed atrophy and apoptosis of the neurons in cerebral cortex and hippocampus. This was associated with neurofibrillary degeneration as well as argyrophilic inclusion. Schwan cell degeneration and nerve fiber demylination were also encountered. The elaboration of lipid peroxidation products, inhibition of antioxidant enzymes and the morphopathological changes were minimized in the Al/Mel treated groups and markedly improved in Al+Mel treated group CONCLUSION: Chronic aluminum exposure in rabbits had dramatic encephalopathic morphopathological lesions. It enhances the lipid peroxidation production and inhibits the SOD enzyme. Melatonin had a good prophylactic effect as an antioxidant in aluminum encephalopathy. PMID- 16264394 TI - Pregnancy complicated by Myasthenia gravis - twelve years experience. AB - OBJECTIVE: The aim of the study was to evaluate the effect of Myasthenia gravis (MG) on the course of pregnancy, labor and delivery, and the condition of neonate. DESIGN AND SETTINGS: Retrospective chart review of pregnant patients with MG hospitalized in tertiary care center over 12 year period was performed. Course of MG before and during pregnancy, mode of delivery, and condition of the newborn were analyzed. RESULTS: There was 42 913 deliveries in the Polish Mother's Research Institute in Lodz during analyzed period. Thirteen patients with MG were identified, giving the incidence of 3/10 000 live births. Mean age of the patients was 28 years, range 18 - 36 years. Average time since diagnosis was 9 years. Eleven women with MG delivered at term (one twin pregnancy), and two babies were born prematurely. In seven patients pregnancy was ended via normal spontaneous vaginal delivery (NSVD), one was forceps extraction, four patients had elective Cesarean Section (CS), and two had emergency CS. Indications for CS were purely obstetrical. Spontaneous vaginal deliveries were speedy. None of the patients had exacerbation of MG during current pregnancy. Mean birth weight of the newborns was 3014,3 g, range 1730 g to 3700 g. Mean Apgar score was 8,7 at 1 minute and 8,9 at 5 minutes. Only one neonate (second twin) developed Transient Neonatal Myasthenia Gravis (TNMG). CONCLUSIONS: Myasthenia gravis did not have significant effect on the course of pregnancy or mode of delivery. In the twin pregnancies it is possible that only one twin will present with the symptoms of TNMG. PMID- 16264395 TI - Cortisol levels and prospective and retrospective memory in humans. AB - OBJECTIVES: The aim of this study was to examine (i) the influence of cortisol on both prospective and retrospective memory performance and (ii) the role of emotional valence in both types of memory. METHODS: Thirty-four male students participated in a memory task, which measured both prospective and retrospective memory performance. Baseline salivary cortisol levels were assessed. RESULTS: Spearman's rank order correlation analysis showed a significant positive correlation between salivary cortisol levels and retrospective memory performance for neutral words. Cortisol levels were not significantly correlated with prospective memory performance for either negative nor neutral words. CONCLUSIONS: The present results indicate chronic cortisol levels are positively associated with retrospective memory at relatively low concentration ranges, but not prospective memory, in healthy young men. Implications for evaluating the beneficial effects of low-dose cortisol treatment on posttraumatic stress disorder is discussed. PMID- 16264396 TI - Reduced brain perfusion and neurocranial shape abnormalities of the temporal regions in patients with Klinefelter syndrome. AB - OBJECTIVES: In patients with Klinefelter syndrome dissocial behaviour, learning difficulties and low intelligence are common. Thus, the aim of our study was to perform brain perfusion studies and cranial cephalometry in 27 cases of Klinefelter syndrome and compare the results with those in a group of 26 healthy subjects. METHODS: Single photon emission tomography (SPECT) was performed after injection of 20 mCi 99mTc-HMPAO and the data from transaxial slices were analyzed for 46 regions of interest in the cerebellar, thalamic, ventricular and parietal planes. Right/ left ratios were calculated and differences above 10 per cent were considered abnormal. Skull radiographs in frontal and lateral view were taken and measurements of the cerebral part were made. RESULTS: SPECT imaging in 27 Klinefelter patients revealed 82 hypoperfusion foci, most frequently in temporal regions, less frequently in temporoparietal and frontal regions, whereas only 11 hypoperfusion foci in 6 of the 26 control subjects were found. Skull radiography revealed the following abnormalities : flattening of the temporal regions, reduced width of the vault, shortening of the anterior cranial fossa and definitely reduced angle of the cranial base; all these anomalies differed significantly from those in the skulls of the control subjects. CONCLUSIONS: The high coincidence of the location on the temporal regions of brain perfusion defects and the neurocranial shape anomalies indicate that an extra X chromosome in Klinefelter patients has detrimental effects on the temporal lobe development and function. PMID- 16264398 TI - Morphological and biochemical study of the pineal gland of pregnant and non pregnant female vizcachas (Lagostomus maximus maximus). AB - OBJECTIVES: The morphological and biochemical aspects of the pineal gland of pregnant and non-pregnant female vizcachas were investigated to examine whether there is a relationship between the pregnancy and this gland. METHODS: Pregnant and non-pregnant adult female vizcachas were used. The ultrastructure of type I cell (dark and light pinealocytes) was studied. Pineal and serum melatonin were investigated by radioimmunoassay. Radiometric methods were applied to investigate the arylalkylamine-N-acetyltransferase (AA-NAT) and hydroxyindole-O methyltransferase (HIOMT) activities. RESULTS: The morphological study showed the existence of dark and light pinealocytes. The differences between both cells are slight. However, the appearance of dark pinealocytes suggests that they have a high activity. These cells were more abundant in the pineal gland of pregnant females. The pineal melatonin levels and the AA-NAT activity were higher in the pregnant females. Serum melatonin and HIOMT activity did not show significant differences. CONCLUSION: Our results suggest that the morphological and biochemical changes in the pineal gland of pregnant vizcachas are associated with the gestation. However, the relation between the pineal gland and the pregnancy is complex and further studies are necessaries to clearly establish it. PMID- 16264397 TI - N-bromoacetyltryptamine strongly and reversibly inhibits in vitro melatonin secretion from mammalian pinealocytes. AB - OBJECTIVES: Cell-permeable and specific inhibitors of melatonin secretion are sill lacking among tools of the pineal research. Recently, a large effort has been made in the development of arylalkylamine N-acetyltransferase inhibitors, but in most cases the new drugs were tested exclusively using cell-free assays or non-pineal cells. The aim of the present study was to characterize the effect of N-bromoacetyltryptamine (BAT), the first synthesized cell-permeable inhibitor of arylalkylamine N - acetyltransferase, on melatonin secretion from rat and pig pineal glands. METHODS: The studies were performed in the superfusion cultures of rat and pig pineal explants. Melatonin secretion was determined by radioimmunoassay (RIA). RESULTS: BAT strongly inhibited the non-stimulated and norepinephrine - stimulated melatonin secretion from the pig and rat pineal explants, with ED50 0.3 - 0.7 microM. The adrenergic stimulation did not modify significantly the inhibitory potency of BAT on the melatonin release. The decline in melatonin secretion induced by the BAT - treatment was biphasic in both rat and pig pinealocytes, with an initial rapid phase followed by a slow one. The half-time of BAT-induced decline in the non - stimulated and norepinephrine stimulated melatonin secretion was ca. 25 - 35 minutes. The inhibitory effect of BAT was reversible in pinealocytes of both investigated mammals. CONCLUSIONS: The results show that BAT is a potent and reversible inhibitor of the melatonin secretion in the mammalian pineal gland and open the way for the use of this inhibitor in investigations on the pinealocyte physiology performed in vitro. PMID- 16264399 TI - Metallothionein stroma reaction in tumor adjacent healthy tissue in head and neck squamous cell carcinoma and breast adenocarcinoma. AB - INTRODUCTION: An accumulation of genetic alterations forming the field of cancerization is an important event for the transformation from normal to cancer cell in multistep carcinogenesis. Histopathologically healthy tumor adjacent tissue might be considered as a cancerization field which is typified by genetic changes required for the development of cancer. Metallothionein (MT) is considered to be a protective and anti-apoptotic protein. The aim of our study was to evaluate the MT expression in head and neck squamous cells carcinoma and breast adenocarcinoma and their histologically healthy adjacent tissue. MATERIALS AND METHODS: We have sampled 29 tissue samples in total derived from head and neck cancers and 29 samples of their clear surgical margins, 33 breast adenocarcinomas and 33 clear surgical margins. Antibody recognizing MT-1 was used for immunohistochemical analysis. RESULTS: MT expression was revealed in 85,7% of head and neck cancers and 94% of breast adenocarcinomas. It was found in all tumor adjacent tissue. MT expression was statistically significantly higher in tumor adjacent tissue than in cancer tissue in cases with the presence of lymph node metastases in both, breast adenocarcinoma and head and neck squamous cell carcinoma. Generally stroma seems to respond to the presence of cancer by the expression of MT, even in tissues which normally do not express MT. CONCLUSIONS: MT might be a normal or protective reaction of healthy adjacent tissue to the presence of tumor. PMID- 16264400 TI - The level of maternal immune tolerance and fetal maturity. AB - OBJECTIVES: Fetal maturity does not seem to be directly connected with the phenomenon of immune tolerance during pregnancy although the fetal maturation influences the process of initiation of the labor at term finishing the immune tolerance during pregnancy. CAP and RCAS1 are expressed by the trophoblast cells and afterwards by the placenta, these proteins are able to modulate the maternal immune response. MATERIALS AND METHODS: 160 patients were randomly selected to our study. The patients were divided into two groups using K score according to the newborn's maturity: maturated and not fully maturated. Within the groups of matured and not fully matured newborns the subgroups were selected according to the type of the labor: spontaneous or induced. The oxytocinase plasma activity was established in plasma samples obtained from pregnant women a few days before delivery. The placental RCAS1 relative amount was assessed by Western blot analysis. RESULTS: The differences in oxytocinase plasma level with respect to the fetal maturity were identified in our study however no RCAS1 expression changes were found regarding the fetal maturation. We determined the alterations in RCAS1 expression with respect to the occurrence of clinical symptoms of the spontaneous beginning of the labor in maturated and not-fully maturated groups of newborns. CONCLUSIONS: Oxytocinase seems to be a useful marker of normal fetal development. The assessment of RCAS1 in placenta directly after delivery appears to indicate the level of maternal immune tolerance during the labor initiation. The level of the immune tolerance at the moment of the delivery drops independently of the fetal maturity. PMID- 16264401 TI - Effects of maternal deprivation on melatonin production and cognition in adolescent male and female rats. AB - OBJECTIVES: It is known that maternal deprivation (MD) may alter cognitive functions such as learning and memory in adult life by effecting normal growth and development. However, the mechanisms of these cognitive alterations are unknown. The aim of this study is to investigate the effects of maternal deprivation on cognition and melatonin production in adolescent male and female rats. METHODS: The litters were separated daily from their mothers for 6 hours on postnatal days 2 to 20. The spatial memory performance was evaluated using a Morris water maze between the postnatal 26th and 32nd days. Plasma melatonin levels were determined on postnatal days 42. RESULTS: MD-rats had longer escape latencies at the second, third and fifth days of training days and spend significantly less time in probe trial, compared to control animals. MAIN FINDINGS: The repeated maternal deprivation caused low blood melatonin levels and there was a significant negative correlation between blood melatonin levels and spatial memory performance in both of male and female adolescent rats. CONCLUSION: These results suggest an association between melatonin production and neurodevelopment. Further studies are needed to determine the interaction between maternal deprivation and pineal gland maturation/function. PMID- 16264402 TI - The effects of zinc deficiency and testosterone supplementation on leptin levels in castrated rats and their relation with LH, FSH and testosterone. AB - AIM: The aim of this study was to investigate how zinc-deficiency and testosterone supplementation, both in combination and individually, affect plasma LH, FSH and leptin levels in castrated rats. DESIGN: Group 1, Control Group. Group 2, Castration Group. Group 3, Testosterone Group. Group 4, Zinc-deficient Group. Group 5, Testosterone, Zinc-deficient Group. Group 6, Zinc-deficient, Castration Group. Group 7, Testosterone, Castration Group. Group 8, Zinc deficient, Testosterone, Castration Group. MEASUREMENTS: Plasma zinc, leptin, LH, FSH, free and total testosterone levels were measured. RESULTS: Group 2 had the highest levels of leptin and LH, besides having the highest FSH levels together with Group 6 (p<0.01). Groups 5 and 8 had the lowest leptin levels (p<0.01). Leptin levels in Groups 4 and 7 were higher than those in Groups 5 and 8, but lower than those in all other groups (p<0.01). LH levels in Group 4 were not different than those in Groups 3, 5 and 8, but significantly lower than those in all other groups (p<0.01). Free and total testosterone levels were higher in Group 4 than in castration groups that were not supplemented testosterone, but were lower in the former than in all others (p<0.01). CONCLUSION: Plasma LH may be more effective than testosterone on plasma leptin and zinc can be an important mediator of the effect LH exercises on leptin. PMID- 16264403 TI - Widespread expression of liver receptor homolog 1 in mouse brain. AB - OBJECTIVES: The distribution of Liver receptor homolog 1 (LRH-1) mRNA was studied in mice brain with the aim to establish whether this nuclear hormone receptor is expressed also in the brain in addition to liver and classical steroidogenic tissues. METHODS: Expression of LRH-1 mRNA in juvenile (30 days old) and adult (60 days old) mouse brain was examined using non-radioactive in situ hybridization with digoxigenin labeled cRNA probes and with RT PCR using specific primers. RESULTS: LRH-1 was strongly expressed throughout the brain. Semiquantitative RT PCR revealed very strong expression of LRH-1 mRNA in cerebrum in comparison to liver and testis, and in situ hybridization revealed that LRH-1 mRNA is uniformly expressed in most brain areas. CONCLUSIONS: LRH-1 is strongly expressed throughout the mouse brain suggesting important roles for this transcription factor, although its precise roles in the CNS remain to be elucidated. PMID- 16264404 TI - Changes in plasma homocysteine levels of rats with experimentally induced hypothyroidism and hyperthyroidism. AB - INTRODUCTION: It is claimed in a limited number of studies carried out on human beings that plasma homocysteine levels increased in hypothyroid patients and decreased in hyperthyroid patients. OBJECTIVE: The aim of this study is to determine total plasma homocysteine, thyroid function tests, vitamin B12, folic acid and lipid levels and to explore the relations among them in rat models with induced hypothyroidism and hyperthyroidism with a view to investigating whether hypothyroid and hyperthyroid rat models could represent human hypothyroidism and hyperthyroidism models. MATERIAL AND METHOD: The study included 30 male Wistar Albino species rats with a mean weight of 200 - 250 g. Rats were randomly divided into 3 groups as 1) hypothyroid group, 2) hyperthyroid group and 3) control group. Hypothyroidism was induced by adding 10 mg/kg/day propylthiouracil to rats' drinking water for 30 days. In order to induce hyperthyroidism, rats were administered 10 microg/100 g L-thyroxin ampule via intraperitoneal route for 10 days. RESULTS: We found that total plasma homocysteine level of the hypothyroid group was significantly lower than those of the control group (p<0.05) and the hyperthyroid group (p<0.001). Total plasma homocysteine level of the hypothyroid group was found insignificantly higher than that of the control group (p>0.05) and significantly higher than that of the hyperthyroid group (p<0.001). We established a significant and positive correlation between total plasma homocysteine level and thyroid hormone levels. We did not identify a significant relation between total plasma homocysteine level and serum folic acid and serum vitamin B12 levels. CONCLUSION: Our findings are different from the findings reported in human hypothyroidism and hyperthyroidism studies. We believe that hypothyroid and hyperthyroid rat models cannot represent human hypothyroidism and hyperthyroidism models. PMID- 16264405 TI - Hyperprolactinemia presenting with encephalomalacia-associated seizure disorder and infertility: a novel application for bromocriptine therapy in reproductive endocrinology. AB - OBJECTIVES: To describe successful oral bromocriptine therapy for hyperprolactinemia accompanied by seizure disorder and encephalomalacia identified during infertility evaluation. MATERIALS & METHODS: A 32 year-old male with an unspecified seizure disorder was referred for infertility consultation. The initial seizure was at age 12, and was treated sequentially with phenytoin, phenobarbitol, and carbamazepine monotherapy which was maintained for six years and then discontinued. At age 26, a second seizure was experienced and 150mg lamotrigine b.i.d. was initiated. When the patient married it was discovered that erection was impossible; tadalafil was administered without success. At our center, total testosterone was low at 107ng/dl and semen analysis revealed oligoasthenozoospermia. Serum prolactin was 79.3ng/ml; visual field examination was normal. Brain MR showed no pituitary enlargement. However, encephalomalacia of the right occiptal lobe with porencephalic dilation of the occipital horn of the right lateral ventricle was present. RESULTS: The patient was initially placed on 2.5mg/d bromocriptine but after three months only a nominal reduction of serum prolactin was achieved. Serum prolactin normalized (25.4ng/ml) after bromocriptine was increased to 7.5mg/d. Repeat semen analysis showed overall sperm concentration of 85M/ml, 30% motility and 12% normal forms morphology (Kruger strict criteria). Total testosterone level was 191ng/dl after final bromocriptine dose was attained, which was well tolerated. DISCUSSION: Bromocriptine has an established role in the management of hyperprolactinemia, particularly in women. For males with elevated serum prolactin even when associated with focal encephalomalacia and seizure disorder, bromocriptine therapy can offer safe benefits including improved semen parameters and normal serum testosterone levels. PMID- 16264406 TI - Activity of estradiol and selective estrogen receptor modulators in the mouse N20.1 oligodendrocyte/astrocytes cell line. AB - OBJECTIVE: The mechanism through which estrogen exerts its neuroprotective and anti-neurodegenerative effects in the central nervous system is poorly understood. Human glial cells are implicated in the pathogenesis of Alzheimer's disease and have both alpha and beta estrogen receptors (ER). We developed a glial cell model for ER function using the N20.1 mouse oligodendroglial cell line to evaluate the response of ERalpha and ERbeta to estradiol (E2), a raloxifene analog LY117018 (LY) and 4-hydroxytamoxifen (4OHT). DESIGN: We tested the ability of exogenous ER to activate transcription in response to ligands (100 nM) using the glial cell line N20.1 in a transient cotransfection assay with an ERalpha or ERbeta expression vector, an ERE-driven reporter and a Renilla luciferase transfection control. RESULTS: Endogenous ER was not detected in the N20.1 cells by Western immunoblotting. E2 stimulated both ERalpha and ERbeta on both ERE- and AP-1 driven promoters. The transcription stimulation by E2 in the ERalpha and ERbeta through the AP-1driven promoter, though significant, was not of the same magnitude as the stimulation of the ERalpha through the ERE-driven promoter. 4OHT and LY did not show significant transcriptional activation of either the ERalpha or ERbeta, through either the ERE or AP-1 driven promoters. LY, at a 10-fold higher concentration than E2, showed a difference in its antagonist activity on the ERbeta through the AP-1 pathway when compared with the ERE- driven promoter, demonstrating not only promoter specificity, but also receptor specificity. CONCLUSIONS: This is the first description of the activity of 4OHT and LY on estrogen receptors in glia. PMID- 16264407 TI - Loss of heterozygosity in 73 human thyroid tumors. AB - OBJECTIVES: The aim of the study was to establish the LOH frequency of selected polymorphic markers in different histological types of thyroid tumors: 18 colloid goiters (CG), five follicular adenomas (FA), nine follicular carcinomas (FTC), 40 papillary carcinomas (PTC), and one anaplastic carcinoma (ATC). For PTC, tumors negative for RET/PTC rearrangements were preferred. METHODS: LOH studies were performed using 14 highly polymorphic markers previously described as frequently lost in thyroid tumors. RESULTS: In 20 cases (27%) the loss of at least one marker was found. No difference between the frequency of the LOH in FTC and PTC tumors was revealed (33% v. 33%). No differences between histopathological subtypes of PTC in LOH were found. Papillary thyroid carcinomas showed a tendency to higher LOH frequency from patients older than 45 years of age compared to younger ones (9/23 v. 4/17) although it was not statistically significant. CONCLUSIONS: We conclude that papillary thyroid cancers, particularly those diagnosed in patients older than 45 years of age, do exhibit LOH at least with the same frequency as follicular cancers. This increased number of LOH events may contribute to the clinical aggressiveness of cancer in older patients. PMID- 16264408 TI - Activation of rat pituitary-adrenocortical and sympatho-adrenomedullary system in response to different stressors. AB - OBJECTIVE: The effects of three different long-term (21 days) stresses: isolation(LTI), forced swimming (LTS) and isolation accompanied by forced swimming (LTI+LTS) on the level of plasma noradrenaline (NA), adrenaline (A), corticosterone (CORT) and adrenocorticotropic hormone (ACTH), both under basal conditions and in response to short-term immobilization and cold as heterotypic additional stressors, were compared. METHODS: Plasma NA and A were assayed by the radioenzymatic method. Plasma CORT was measured using RIA kits. Plasma ACTH was determined by chemiluminescent method. RESULTS: LTI produced a significant elevation of basal plasma CORT and ACTH, while basal plasma NA and A concentrations remained unchanged. Combination of long-term isolation and forced swimming, produced a significant elevation of basal plasma ACTH content only, while LTS did not influence the basal level of this hormone. When LTI rats were exposed to immobilization and cold, a significant elevation of plasma NA and A level was recorded. In LTS and LTI+LTS groups of rats exposed to immobilization or cold, increased plasma NA and A levels were observed, but this increase was lower in comparison with that found in LTI rats. No difference in plasma CORT content between the three long-term stressed groups of animals was observed, while plasma ACTH level was significantly more elevated in LTS and LTI+LTS than in LTI rats. CONCLUSION: Based on these results, it may be concluded that LTI as a psychosocial stress represents a stronger stressor than LTS. Also, daily short term (15 min, 21 days) swimming stress seems to attenuate the effect of long-term isolation on the activity of sympatho-adrenomedullary system. PMID- 16264409 TI - Hyperplasia of the colonic neuroendocrine cells after pinealectomy in rats. The new evidence for the existence of connections between the distant parts of the diffuse neuroendocrine system. AB - OBJECTIVES: The diffuse neuroendocrine system (DNES) participates in the systemic homeostasis and may work as a unified system with integrated functions but anatomically disperse. However, the mechanisms that are involved in the integration of the distant parts of the DNES are poorly known. This study is aimed on the effects of a pinealectomy on the population of the neuroendocrine cells (NECs) in the colon of rats. METHODS: A group of seven newborn rats and one of adult rats were submitted to a pinealectomy. The animals were sacrificed after 15 days and 90 days, respectively. The control groups of adult and newborn rats were subjected to a 'sham surgery' and then sacrificed after similar periods of time. Paraffin-included sections of the colon samples were stained by the Churukian-Schenk method in order to selectively stain the NECs. The NEC index was estimated by the expression: iNEC = NECs population per crypt/Total cell population of the crypt. The results were expressed as median and absolute range, and the statistical significance was demonstrated by the Mann-Whitney (U) test. RESULTS: The morphometric analysis showed a significant increase in the number of colonic NECs of the adult rats subjected to a pinealectomy when compared to the controls. No statistical difference was observed between the iNECs of the newborn groups of rats. CONCLUSION: This finding suggests the existence of regulatory mechanisms that integrate distant parts of the DNES, by adapting the population of the endocrine cells after the ablation of an important component of the system. Further studies on the mechanisms involved in the integration of the DNES may be facilitated by the simple experimental model that we propose. PMID- 16264410 TI - The effects of periodic alteration of the temperature on the rhythmic melatonin release of explanted chicken pineals. AB - The melatonin rhythm of cultured chicken pineal cells can be synchronized by cyclic environmental effects. Unlike the effects of light on the melatonin secretion, those of the temperature changes are much less known. Similarly, only a few data are available on the interactions between environmental illumination and periodic temperature changes and on the sensitivity of the pineal gland to temperature changes in different ages of animals. We monitored the effects of temperature on chicken pineals for several days in vitro, in a perifusion system under different illumination patterns. The effects of temperature on pineals from chicken of different age were also compared. The phase of the melatonin rhythm was controlled by periodic elevations of temperature under both constant darkness and continuous illumination. These results show that rhythmic changes of temperature prevent desynchronization induced by constant light. Following elevation of the temperature, the melatonin rhythm of pineals of young chickens (less, than 14 weeks old) was altered for 16 - 18 hours. Similar changes in melatonin rhythm were not found in older animals. It is concluded that the sensitivity for temperature changes of the pineal cells is varying with age. PMID- 16264411 TI - Familial occurrence of adrenocortical insufficiency in two brothers with Allgrove syndrome. A case report of 4A (Allgrove) syndrome with epilepsy and a new AAAS gene mutation. AB - Allgrove syndrome is a rare autosomal recessive disease with achalasia, alacrima, adrenocortical insufficiency, autonomic neuropathy and other neurological disturbances. A case of two brothers with Addison s disease from early childhood is presented. The younger brother with Addison disease died at the age of 5. The older brother was treated for adrenocortical insufficiency from the age 3, and then treated for achalasia and epilepsy from the age of 5. The patient is currently 26 years old and suffers from achalasia and adrenocortical insufficiency. He also suffers from alacrima, autonomic neuropathy, epilepsy and other damages of the central and peripheral nervous system. The clinical picture is typical for Allgrove or 4A syndrome, and the diagnosis was confirmed by means of molecular analysis of a new AAAS gene mutation. PMID- 16264412 TI - Mercury and autism: accelerating evidence? AB - The causes of autism and neurodevelopmental disorders are unknown. Genetic and environmental risk factors seem to be involved. Because of an observed increase in autism in the last decades, which parallels cumulative mercury exposure, it was proposed that autism may be in part caused by mercury. We review the evidence for this proposal. Several epidemiological studies failed to find a correlation between mercury exposure through thimerosal, a preservative used in vaccines, and the risk of autism. Recently, it was found that autistic children had a higher mercury exposure during pregnancy due to maternal dental amalgam and thimerosal containing immunoglobulin shots. It was hypothesized that children with autism have a decreased detoxification capacity due to genetic polymorphism. In vitro, mercury and thimerosal in levels found several days after vaccination inhibit methionine synthetase (MS) by 50%. Normal function of MS is crucial in biochemical steps necessary for brain development, attention and production of glutathione, an important antioxidative and detoxifying agent. Repetitive doses of thimerosal leads to neurobehavioral deteriorations in autoimmune susceptible mice, increased oxidative stress and decreased intracellular levels of glutathione in vitro. Subsequently, autistic children have significantly decreased level of reduced glutathione. Promising treatments of autism involve detoxification of mercury, and supplementation of deficient metabolites. PMID- 16264413 TI - Demonstration of additivity failure in human circadian phototransduction. AB - OBJECTIVES: Published data, both on nocturnal melatonin suppression in humans and on widely accepted retinal structure and function, suggest that spectral opponency plays a role in human circadian phototransduction. We directly test subadditivity, implied by spectral opponency, in human circadian phototransduction in response to nearly monochromatic and to polychromatic light. METHODS: Adult male human subjects were exposed for 60 minutes to two intensities each of two lighting conditions, during nighttime experimental sessions. One condition consisted of light from mercury vapor lamps (450 and 1050 lx), and one condition consisted of light from these lamps filtered such that only the spectral line from this lamp at 436 nm was presented to subjects (7.5 and 15 lx). RESULTS: Melatonin suppression from the filtered illumination at 436 nm alone was greater than mercury lamp illumination (containing energy at 436 nm in addition to other wavelengths), even when the sources exposed subjects' retinae to equal amounts of irradiance at 436 nm. CONCLUSION: This direct test of subadditivity, together with evidence from neuroanatomy, supports the inference that spectral opponency is a fundamental characteristic of human circadian phototransduction. PMID- 16264414 TI - Decreased dehydroepiandrosterone sulfate but normal insulin-like growth factor in chronic fatigue syndrome (CFS): relevance for the inflammatory response in CFS. AB - There are a few reports that chronic fatigue syndrome (CFS) may be accompanied by changes in hormones, such as dehydroepiandrosterone (DHEA) and insulin-like growth factor (IGF1). This study examines the serum concentrations of DHEA sulfate (DHEAS), IGF1 and IGF1 binding protein-3 (IGFBP3) in 20 patients with CFS and in 12 normal controls. The IGFBP3/IGF1 ratio was computed as an index for IGF1 availability. We found significantly lower serum DHEAS concentrations in CFS, but no significant differences either in IGF1 or the IGFBP3/IGF1 ratio between CFS patients and normal controls. The decrease in serum DHEAS was highly sensitive and specific for CFS. There were significant and positive correlations between serum DHEAS and serum zinc and the mitogen-induced expression of the CD69 molecule on CD3+CD8+ T cells (an indicator of early T cell activation). There was a significant and negative correlation between serum DHEAS and the increase in the serum alpha-2 protein fraction (an inflammatory marker). Serum IGF1, but not DHEAS, was significantly and inversely correlated to age. The results show that CFS is accompanied by lowered levels of DHEAS and that the latter may play a role in the immune (defect in the early activation of T cells) and the inflammatory pathophysiology of CFS. PMID- 16264415 TI - Decreased level of novelty seeking in blood donors infected with Toxoplasma. AB - OBJECTIVES: Toxoplasma gondii, a parasitic protozoan, infects about 30 - 60% of people worldwide. Toxoplasma is known to induce behavioral changes and an increase of dopamine in mice. The presence of anti-Toxoplasma antibodies (latent toxoplasmosis) is also a risk factor for schizophrenia. Latent toxoplasmosis in men (male soldiers) is associated with lower novelty seeking. As the novelty seeking is supposed to negatively correlate with level of dopamine, the observed effect was interpreted as indirect evidence of increased dopamine levels in subjects with toxoplasmosis. However, it is also possible that the observed effect was caused by association of both novelty seeking and Toxoplasma infection with a third factor, e.g. size of place of residence. METHODS: Personality profile of 290 blood donors (205 men and 85 women) were measured by Cloninger's TCI (Temperament and Character Inventory) and their blood samples were assayed for the presence of anti-Toxoplasma antibodies. Difference between Toxoplasma infected and Toxoplasma-free subjects was tested with ANCOVA method with gender, size of place of residence, and age as covariates. RESULTS: The present analysis revealed that lower novelty seeking was associated with latent toxoplasmosis both in men and women. The effect of infection on novelty seeking remained significant even after adjustment for size of place of residence (p<0.01). CONCLUSION: Decreased novelty seeking in Toxoplasma-infected subjects have been already confirmed in three independent populations (male soldiers and male and female blood donors). These findings suggest that the local inflammation-induced increase in dopamine in the brain of infected subjects can represent a missing link between toxoplasmosis and schizophrenia. PMID- 16264416 TI - Relation of acute-phase reaction and endothelial activation to insulin resistance and adiposity in obese children and adolescents. AB - OBJECTIVES: There is increasing evidence that an ongoing cytokine-induced acute phase response is closely involved in the pathogenesis of type 2 diabetes and associated complications such as dyslipidemia and atherosclerosis. The aim of this study was to investigate the relationship of inflammation and endothelial activation with insulin resistance in childhood obesity. METHODS: Two hundred and eleven (122 boys) obese children and adolescents were examined. Fasting levels of ultra-sensitive C-reactive protein (CRP), fibrinogen (FB), interleukin-6 (IL-6), interleukin-1beta (IL-1beta), intercellular cell adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), von Willebrand factor (vWF), glucose, insulin, and HbA1c were determined. Insulin resistance was assessed by the homeostasis method. RESULTS: HOMA IR correlated significantly with all measures of adiposity as well as with majority of inflammation and endothelial dysfunction markers. After adjustment for age, gender, BMI and fat mass, the correlation with insulin resistance remained significant for CRP, ICAM-1 and von Willebrand factor. There was a trend for association between HOMA IR and IL-6 as well as HOMA IR and fibrinogen. CONCLUSION: Acute-phase reaction and endothelial activation correlate with insulin resistance in obese youth. It is possible that the cluster of these pro-atherogenic factors may contribute to the accelerated atherosclerosis in obese children. PMID- 16264417 TI - Galanin modulates pituitary hormones release. AB - OBJECTIVES: Galanin and its receptors are widely distributed within the central and peripheral nervous system, especially in hypothalamus including preoptic area, paraventricular nucleus (PVN), supraoptic nucleus (SON) and median eminence. Galanin plays an important role in the control of food intake, energy expenditure, reproduction, water balance and various neuroendocrine functions. Galanin may affect hormones release, but the exact mechanism of the peptide action remains unclear and possible direct effects of galanin on the pituitary are controversial. The aim of this study was to examine the effects of galanin on pituitary hormones release after the central and peripheral administration of the peptide. MATERIAL AND METHODS: (i) Experiment I - Intracerebroventricular (icv) administration of galanin: Galanin at a concentration of 0.5 microg in 5microl vehicle (artificial cerebrospinal fluid) or equal volume of the vehicle was slowly (1microl/min) infused into the third ventricle with an automatic pump (CMA/100; Sweden) through an inner cannula inserted into the guide cannula. After the end of the infusion the rats were transferred to their home cages with free access to food and water. At 60 min after the infusion of galanin or vehicle, animals were decapitated and trunk blood was collected in plastic tubes containing 1000 IU aprotinin (inhibitor of protease) per each ml of blood. (ii) Experiment II - Intravenous (iv) injection of galanin: Galanin in a dose of 10 microg in 300 microl of saline or 300 microl of saline alone was injected into the tail vein. After the injection the animals were transferred to individual cages with free access to food and water. At 60 min after the injection of galanin or saline, animals were decapitated, and trunk blood was collected in plastic tubes containing 1000 IU of aprotinin (Trascolan). The blood samples were centrifuged (3000 rpm for 20 min at 4 degrees C). Serum samples were frozen until hormonal analyses were performed. Serum rLH, rFSH, rPRL, rGH, rTSH concentrations were measured with RIA methods. RESULTS: Galanin administered icv increased significantly rPRL and rTSH levels (p<0.01, p<0.05, respectively). Plasma rPRL and rTSH concentrations were not changed after iv injection of galanin. Galanin injected centrally inhibited significantly rGH release (p<0.01), however, galanin given iv stimulated rGH (p<0.01). Serum rLH and rFSH concentrations were not changed after icv and iv injections of galanin. CONCLUSION: Galanin may be involved in the modulating mechanism of pituitary hormones release. PMID- 16264418 TI - Growth hormone (GH) secretion and pituitary size in children with short stature. Efficacy of GH therapy in GH-deficient children, depending on the pituitary size. AB - INTRODUCTION: Certain relationships between pituitary size and growth hormone (GH) secretion have previously been observed, however they are still a matter of controversy. Organic abnormalities of the hypothalamic-hypophyseal region are important for predicting growth response to GH therapy. AIM: Evaluation of relations between GH secretion and the pituitary size in short children and estimation of the efficacy of GH therapy in children with GH deficiency (GHD). PATIENTS AND METHODS: The analysis comprised 216 short children (159 boys). Two GH stimulation tests, as well as magnetic resonance image (MRI) examination, were performed in each patient. All the patients with GHD were treated with GH for, at least, one year. RESULTS: Significant correlations were found between pituitary height and GH secretion (p < 0.05). Patients were classified into three (3) groups: 1) pituitary hypoplasia (HP) for height age; 2) HP for the chronological age but not for the height age; 3) normal pituitary size. Significant differences in GH secretion were observed among the groups (6.1+/-5.3 vs. 8.1+/-4.4 vs. 12.3+/-9.1 ng/mL, respectively). There was a negative correlation between GH peak and height gain during GH therapy (r = -0.34). The highest growth improvement was noticed in patients with HP for the height age. CONCLUSIONS: Pituitary hypoplasia for the height age is related to more severe GH deficiency and the best response to GH therapy. PMID- 16264419 TI - Family matters. PMID- 16264420 TI - Like it or not, life-extension research extends beyond biogerontology. PMID- 16264421 TI - Human agency in the neurocentric age. Philosophers and scientists resort to dualistic explanations to reconcile the age-old dichotomy between determinism and 'free will', but agency is an integral part of human biology. PMID- 16264422 TI - Science fact and the SENS agenda. What can we reasonably expect from ageing research? PMID- 16264423 TI - Consciousness is missing--and so is research. After the Schiavo controversy in the USA, obstacles still hinder the study of people with little or no awareness. PMID- 16264424 TI - Where the future went. In response to market trends and patenting laws, genomics companies are adapting their strategies. PMID- 16264425 TI - The squeaky wheel gets the grease. Research funding is not necessarily allocated to those who need it most. PMID- 16264426 TI - Membrane protein insertion: mixing eukaryotic and prokaryotic concepts. AB - Proteins are translocated across or inserted into membranes by machines that are composed of soluble and membrane-anchored subunits. The molecular action of these machines and their evolutionary origin are at present the focus of intense research. For instance, our understanding of the mode of insertion of beta-barrel membrane proteins into the outer membrane of endosymbiotically derived organelles has increased rapidly during the past few years. In particular, the identification of the Omp85/YaeT-involving pathways in Neisseria meningitidis, Escherichia coli and cyanobacteria, and homologues of Omp85/YaeT in chloroplasts and mitochondria, has provided new clues about the ancestral beta-barrel protein insertion pathway. This review focuses on recent advances in the elucidation of the evolutionarily conserved concepts that underlie the translocation and insertion of beta-barrel membrane proteins. PMID- 16264427 TI - The chromosome cycle: coordinating replication and segregation. Second in the cycles review series. AB - During the cell-division cycle, chromosomal DNA must initially be precisely duplicated and then correctly segregated to daughter cells. The accuracy of these two events is maintained by two interlinked cycles: the replication licensing cycle, which ensures precise duplication of DNA, and the cohesion cycle, which ensures correct segregation. Here we provide a general overview of how these two systems are coordinated to maintain genetic stability during the cell cycle. PMID- 16264433 TI - Drug therapy for atrial fibrillation: where do we go from here? AB - Atrial fibrillation, the most common cardiac arrhythmia requiring medical attention, has effects that range from mild symptoms to devastating stroke. Although treatments have evolved since the foxglove plant (later identified as containing digitalis) was first administered to slow the heart rate, satisfactory drug therapy has not been developed. In this review we describe present-day medical options and developments of future therapies to treat atrial fibrillation and maintain normal sinus rhythm. PMID- 16264428 TI - Functional elements residing within satellite DNAs. AB - Satellite DNAs represent a fast-evolving portion of the eukaryotic genome whose evolution is proposed to be driven by the stochastic process of molecular drive. Recent results indicate that satellite DNAs are subject to certain structural constraints, which are probably related to their interaction with proteins involved in the establishment of specific chromatin structures. The evolutionary persistence and high sequence conservation of some satellites, as well as the presence of stage- or tissue-specific, differentially expressed transcripts in several species, are consistent with the hypothesis that satellite DNA could have a regulatory role in eukaryotic organisms. Although the role of most transcripts is not known, some act as precursors of small interfering RNAs, which are now recognized as having an important role in chromatin modulation and the control of gene expression. Furthermore, some transcripts are involved in the cellular response to stress. PMID- 16264434 TI - Combinatorial pharmacogenetics. AB - Combinatorial pharmacogenetics seeks to characterize genetic variations that affect reactions to potentially toxic agents within the complex metabolic networks of the human body. Polymorphic drug-metabolizing enzymes are likely to represent some of the most common inheritable risk factors associated with common 'disease' phenotypes, such as adverse drug reactions. The relatively high concordance between polymorphisms in drug-metabolizing enzymes and clinical phenotypes indicates that research into this class of polymorphisms could benefit patients in the near future. Characterization of other genes affecting drug disposition (absorption, distribution, metabolism and elimination) will further enhance this process. As with most questions concerning biological systems, the complexity arises out of the combinatorial magnitude of all the possible interactions and pathways. The high-dimensionality of the resulting analysis problem will often overwhelm traditional analysis methods. Novel analysis techniques, such as multifactor dimensionality reduction, offer viable options for evaluating such data. PMID- 16264435 TI - New concepts in drug discovery: collateral efficacy and permissive antagonism. AB - New perspectives on the complexity of G-protein-coupled receptor (GPCR) signalling and the increased resolution of existing tools for studying GPCR behaviour has led to the conception of new hypotheses that affect the discovery of drugs acting at GPCRs. Taking into consideration the novel concepts of collateral efficacy and permissive antagonism in the search for synthetic agonists and antagonists, respectively, will be essential in the search for drugs with unique therapeutic profiles. Here, the design of drugs against HIV is used as an example of how these concepts might be taken into consideration for GPCR targeted drugs in general. PMID- 16264770 TI - Testosterone replacement therapy for late onset hypogonadism: what is the risk of inducing prostate cancer? AB - Prescription sales of testosterone have risen considerably over the last decade and are likely to continue to grow as further preparations become available. Testosterone promotes existing prostate cancer; however, concern does exist as to whether or not testosterone therapy induces prostate cancer. The aim of this article is to review the evidence for such a link. PMID- 16264436 TI - Acyclic nucleoside phosphonates: a key class of antiviral drugs. AB - Almost 20 years after the broad antiviral activity spectrum of the first acyclic nucleoside phosphonates was described, several of these compounds have become important therapies for DNA virus and retrovirus infections. Here, we review the discovery and development of acyclic nucleoside phosphonates, focusing on cidofovir and its potential in the treatment of various herpes-, papilloma-, polyoma-, adeno- and pox-virus infections, adefovir for the treatment of hepatitis B and tenofovir for the treatment of AIDS and the prevention of HIV infections. PMID- 16264771 TI - Controlled-release doxazosin in the treatment of benign prostatic hyperplasia. AB - PURPOSE: To evaluate the effectiveness and safety of controlled-release doxazosin for benign prostatic hyperplasia (BPH). SCOPE: In this open-label, noncontrolled, observational surveillance study, 3684 men with BPH received 4-8 mg of controlled release doxazosin gastrointestinal therapeutic system (GITS) for 6 months; 3283 (89.1%) patients completed the trial. Changes in urinary symptoms and quality of life were assessed using the International Prostate Symptom Score (IPSS). Blood pressure and adverse events were assessed. CONCLUSION: After 6 months' treatment with doxazosin GITS resulted in significant improvements in IPSS. BP was reduced only in hypertensive patients. Doxazosin, GITS was well tolerated. PMID- 16264773 TI - The impact of epidermal-growth-factor-receptor mutations in response to lung cancer therapy. PMID- 16264774 TI - When is a genomic classifier ready for prime time? PMID- 16264789 TI - Can combined whole brain radiation therapy and radiosurgery improve the treatment of single brain metastases? PMID- 16264790 TI - Does concurrent postoperative radiotherapy and chemotherapy in head and neck cancer improve patient outcome? PMID- 16264791 TI - Can dexrazoxane reduce myocardial injury in anthracycline-treated children with acute lymphoblastic leukemia? PMID- 16264792 TI - Are symptoms reliable as an indication for the necessity of bone scans in lung cancer metastases? PMID- 16264793 TI - Is fulvestrant more effective than tamoxifen for treating ER-positive breast cancer in postmenopausal women? PMID- 16264794 TI - Can EPO reduce blood transfusion requirements during induction therapy for high risk neuroblastoma? PMID- 16264795 TI - Can exemestane improve adjuvant treatment for postmenopausal women with primary breast cancer? PMID- 16264796 TI - Long-term survivors of childhood cancers: what knowledge have we gained? AB - Currently, in the US and Western Europe about 75% of children diagnosed with cancer survive at least 5 years and most survivors are cured. Unfortunately, survivors are at an increased risk of a range of adverse health outcomes compared with that expected from the general population. Large-scale studies of mortality occurring beyond 5-year survival show that the majority of the deaths in the initial decade are because of recurrent tumors. Thereafter, other causes of death that occur increasingly including second malignant neoplasms, cardiac deaths and pulmonary deaths. During the past 25 years, research has focused on estimating the risks of such long-term 'effects' of childhood cancer and its treatment. Studies have examined how risks vary in relation to a range of factors that include treatment exposures, age at treatment, type of childhood cancer, duration of follow-up, and factors that may genetically predispose a survivor to particular complications. This information is critical to counsel survivors and their families in relation to long-term risks and identify specific groups of survivors who are at particularly high risk--with a view to earlier diagnosis or intervention and provide the means of assessing potential benefits and risks of different proposals for future treatment protocols. PMID- 16264797 TI - Allogeneic hematopoietic stem-cell transplantation: the next generation of therapy for metastatic renal cell cancer. AB - The management of metastatic renal cell carcinoma (mRCC) remains a therapeutic challenge; less than 10% of patients survive for longer than 5 years. The resistance of renal cancer to chemotherapy may be explained by high levels of the multidrug resistance gene, MDR1. Immune-based treatments for renal cancer have been explored because of their unusual susceptibility to immunological assault. However, response rates to cytokines such as interleukin-2 and interferon-alpha have ranged from only 10% to 20%, prompting other immunotherapy approaches, such as allogeneic stem-cell transplantation, to be investigated. Several clinical trials have provided evidence of partial or complete disease regression in refractory mRCC following nonmyeloablative stem-cell transplantation. This effect is because of a donor antimalignancy effect mediated by immunocompetent donor T cells, called graft-versus-tumor effect. Unfortunately, less than 30% of patients who could have this procedure will have a human-leukocyte-antigen-compatible sibling, and attention is focusing on alternative donors such as matched unrelated donors and partially mismatched related donors. Despite the improved safety of nonmyeloablative conditioning regimens, transplant-related toxic effects (particularly graft-versus-host disease) remain obstacles to the safe and effective use of this treatment. Regardless of these limitations, innovative approaches have attempted to harness the potential of the graft-versus-tumor effect in mRCC and other solid tumors. PMID- 16264798 TI - Targeting angiogenesis in cancer: clinical development of bevacizumab. AB - The importance of vascular endothelial growth factor (VEGF) as a regulator of normal and tumor blood vessel growth has been increasingly characterized over the past two decades. VEGF increases vascular permeability and has a well established role in stimulating angiogenesis, a prerequisite of tumor growth. Numerous compounds have been developed to counteract the angiogenic effects of VEGF. One such drug, bevacizumab, a humanized anti-VEGF monoclonal antibody, received FDA approval in the US earlier this year. Results obtained from initial trials showed that this drug was generally well tolerated, and combination studies of bevacizumab and chemotherapeutic agents have been completed in patients with breast, prostate, lung, and colorectal cancers. A randomized trial of bevacizumab used in combination with capecitabine for metastatic breast patients showed no overall improvement of progression-free survival. However, this result contrasted greatly with the data obtained for patients with advanced colorectal cancer, where a combination regimen of bevacizumab, 5-fluorouracil, leucovorin and irinotecan demonstrated a significant improvement in response rates and overall survival compared with chemotherapy alone. This review highlights the key clinical trial data with bevacizumab and discusses the reasons for some of the contrasting results seen in different patient studies. PMID- 16264799 TI - Technology insight: Emerging techniques to predict response to preoperative chemotherapy in breast cancer. AB - During the past decade, several high-throughput analytical methods have been developed, and most of these are being explored as potential diagnostic tools. Gene expression profiling with DNA microarrays or with multiplex polymerase chain reaction are the methods closest to being of clinical use. Prediction of clinically meaningful response to particular chemotherapy regimens or drugs remains a persistent challenge. There are established clinical and histopathologic predictors of prognosis for breast cancer, but there is no test to assist in selecting the optimal chemotherapy regimen for patients. Here we review recent advances in the application of gene expression profiling to chemotherapy response prediction. PMID- 16264800 TI - Axillary swelling and a reduced general condition in a middle-aged man. AB - BACKGROUND: A 48-year-old man presented to his GP with an indolent swelling in his left axillary region. He also complained of a cough, a feeling of pressure in his chest, general poor health, and had increasingly suffered from night-time sweating and fever. Physical examination and an ultrasound revealed an enlarged lymph node of almost 4 cm in the left axillary region and several smaller lymph nodes of 1.5-2.0 cm in the left cervical region. Laboratory tests were unremarkable, except for an elevated erythrocyte sedimentation rate. INVESTIGATIONS: Excisional biopsy, radiography, ultrasound, CT scan, bone marrow biopsy, radionuclide imaging, echocardiography and lung function tests. DIAGNOSIS: Early-stage unfavorable (intermediate) lymphocyte-rich classical Hodgkin's lymphoma. MANAGEMENT: Chemotherapy (treatment with doxorubicin, bleomycin, vinblastine and dacarbazine) and involved-field radiotherapy. PMID- 16264801 TI - The 'War on Cancer' and its impact. PMID- 16264802 TI - Preoperative radiotherapy in rectal cancers: why a standard in Europe and not in the US? PMID- 16264803 TI - Adjuvant chemotherapy in rectal cancers: why a standard in the US and not in Europe? PMID- 16264818 TI - Biomarkers of response to gefitinib in non-small-cell lung cancer. PMID- 16264819 TI - Can weight loss at presentation predict patient outcome in lung cancer? PMID- 16264821 TI - Can frequency and severity of symptoms predict the presence of ovarian cancer? PMID- 16264822 TI - Are circulating tumor cells an independent prognostic factor in patients with high-risk melanoma? PMID- 16264820 TI - First-line treatment options for patients with metastatic colorectal cancer. PMID- 16264823 TI - Is laparoscopically-assisted colectomy an acceptable operation for colon cancer? PMID- 16264824 TI - Oxaliplatin-related neurotoxicity: is chelation the solution? PMID- 16264825 TI - Mechanisms of disease: Radiosensitization by epidermal growth factor receptor inhibitors. AB - The epidermal growth factor receptor (EGFR) inhibitors are among the most intensely studied new molecular therapeutic agents. Although response rates have been somewhat disappointing when EGFR inhibitors are used as single-agent therapy for advanced disease, these inhibitors may be more effective as chemo- and radiosensitizers. The first phase III randomized trial evaluating EGFR inhibitors as radiosensitizers in patients with locally advanced head and neck cancer was strongly positive, indicating significant potential of this class of agents to improve outcome with radiotherapy. However, optimal implementation of EGFR inhibitors as radiosensitizers depends, in part, on a better understanding of the mechanisms of radiosensitization. Preclinical studies provide important observations with regard to potential mechanisms. The phenotypic cellular changes associated with EGFR inhibition are impressively consistent between different model systems, with almost all studies showing that EGFR inhibitors affect proliferation, angiogenesis, and cell survival. Whether EGFR inhibitors influence response to radiation directly, or whether the improved response is a result of additive effects of the two modalities, remains unclear. However, cell-cycle arrest, endothelial cell sensitivity, and apoptotic potential are all important factors in radiation response of epithelial tumors. Furthermore, less-studied effects of EGFR inhibitors on DNA repair suggest that modulation of DNA damage response to cytotoxic injury might result in radio- or chemosensitization. This review will explore potential mechanisms of radiosensitization by EGFR inhibitors. PMID- 16264826 TI - Drug insight: Cancer cell immortality-telomerase as a target for novel cancer gene therapies. AB - Rapid advances in our understanding of the molecular basis of cancer development and progression over the past three decades have led to the design of new potential cancer therapies. High throughput target validation and expression studies are expected to yield a powerful arsenal of new cancer treatments, but untangling the complex pathways underlying the major cancer phenotypes remains a significant challenge. A considerable body of evidence in recent years implicates deregulated expression of a single multi-component enzyme, telomerase, as a causative factor at the heart of immortalization in the vast majority of human tumors. This review highlights the potential of telomerase as a target for novel cancer therapies. The potential of exploiting the selectivity of the telomerase family of genes within cancer cells to develop gene therapy strategies is discussed, and the progress towards translating these novel therapeutics from the laboratory to the clinic is reviewed. PMID- 16264827 TI - Technology insight: Proton beam radiotherapy for treatment in pediatric brain tumors. AB - Tumors of the central nervous system are the most common solid tumor in childhood. Treatment options for childhood brain tumors include radiation therapy, surgery and chemotherapy, often given in combination. Radiation therapy regularly has a pivotal role in treatment, and technological advancements during the past quarter of a century have dramatically improved the ability to deliver radiation in a more focused manner. Improvements in imaging and computing ability led to better targeting of tumor tissue using conventional X-ray therapy. These advances have been harnessed for proton radiation therapy. Proton radiotherapy has special physical characteristics that allow normal tissues to be spared better than even the most conformal photon radiation, and it will reduce the complications from treatment. This review discusses the characteristics of proton radiation, and describes examples of pediatric brain tumor patients who would benefit most from this form of treatment. PMID- 16264829 TI - The curse of the cure. PMID- 16264828 TI - Technology insight: Identification of biomarkers with tissue microarray technology. AB - High-throughput technologies have been developed in the hope of increasing the pace of biomedical research, and accelerating the rate of translation from bench to bedside. Using such technology in target discovery has resulted in the need for systematic validation of the targets in an equally rapid manner. For example, gene expression microarrays have highlighted many potential targets in cancer, and tissue microarrays have emerged as a powerful tool to validate these targets by measuring tumor-specific protein expression and linking it to clinical outcome. Automated quantitative analysis of the tissue microarray 'spots' is beginning to take the technology a step further, removing observer bias, and providing standards for quality control and the potential for high-throughput analysis. The validation required for translation of tissue biomarkers from the research lab to the clinical lab will probably rely heavily on the combination of tissue microarray technology with automated quantitative analysis. PMID- 16264830 TI - Problems and promises of targeted therapy for Hodgkin's lymphoma. PMID- 16264831 TI - What is the role of radiotherapy in bladder-preserving cancer therapy? PMID- 16264846 TI - Is 6 months of androgen suppression therapy plus radiotherapy of benefit in patients with localized prostate cancer? PMID- 16264847 TI - Can plasma Epstein-Barr virus DNA levels be used to monitor nasopharyngeal carcinoma progression? PMID- 16264848 TI - How do personal characteristics affect sensitivity and specificity of mammography? PMID- 16264849 TI - Should patients with acute lymphoblastic leukemia receive hematopoietic stem-cell transplant from an unrelated donor? PMID- 16264851 TI - Appropriate use of surveillance colonoscopy after polypectomy. PMID- 16264850 TI - Can the addition of cetuximab to irinotecan improve outcome in colorectal cancer? PMID- 16264852 TI - Can a computer-based system be used to educate women on genetic testing for breast cancer susceptibility? PMID- 16264854 TI - Intensity-modulated radiation therapy in the treatment of head and neck cancer. AB - Intensity-modulated radiation therapy (IMRT) is a new technical improvement of radiotherapy, in which computer-controlled treatment machines produce multiple beams of radiotherapy whose intensity is optimized to deliver a high dose of radiation to specified volumes, while reducing the dose to adjacent non-target organs. The potential benefits include the ability to deliver higher doses to the target with an improved safety than has previously been possible, and to reduce side effects and complications. Using IMRT to treat some head and neck cancers is especially attractive due to the close vicinity of the targets and many critical, dose-limiting and non-involved structures, and because of the lack of breathing related motion. The main clinical uncertainties in the use of IMRT for head and neck cancer relate to uncertainties in the extent of radiation to the target areas. In addition, large volumes of adjacent, non-target tissue receive moderate to low radiation doses, raising concerns of increased risk of radiation-related carcinogenesis in young patients. Initial promising clinical data have emerged from IMRT treatment of several head and neck tumor sites. PMID- 16264853 TI - Genomics in breast cancer-therapeutic implications. AB - The introduction of DNA microarray techniques has had dramatic implications on cancer research, allowing researchers to analyze expression of multiple genes in concert and relate the findings to clinical parameters. The main discoveries in breast cancer, as well as in other malignancies, have so far been with respect to two key issues. First, individual tumors arising from the same organ may be grouped into distinct classes based on their gene expression profiles, independent of stage and grade. Second, the biologic relevance of such classification is corroborated by significant prognostic impact. We review how the use of microarray technologies can provide unique possibilities to explore the mechanisms of tumor behavior in vivo that will allow evaluation of prognosis and, potentially, drug resistance. However, in spite of recent advances, we are not yet at a stage where the use of these techniques should be implemented for routine clinical use, whether to define prognostic factors or to predict sensitivity to therapy. PMID- 16264855 TI - Partial-breast treatment for early breast cancer: emergence of a new paradigm. AB - Although hailed as a paradigm shift, the breast conservative treatment that emerged in the 1980s was in fact an extension of the Halstedian concept, wherein whole-breast irradiation (WBI) compensated for the limited surgery. Observations that 80-90% of breast recurrences after breast conservative surgery and WBI occur in the tumor bed questions the need for protracted elective WBI, and provides the rationale for accelerated-partial-breast irradiation (APBI) of small cancers without adverse features predisposing to multicentric recurrence. APBI can be given over a week with various external beam, intraoperative or brachytherapy (interstitial or MammoSite) techniques. Since the approval of MammoSite by the US FDA in May 2002, a surge of interest has been evident, with 4,000 cases treated using this technique in the past 2 years. Several phase II APBI brachytherapy studies show that 4 to 7-year breast control rates (95%), survival and cosmetic outcome obtained from more than 600 appropriately selected women are comparable to matched or historic controls receiving WBI. The 2 to 3-year interim results of two ongoing randomized trials do not show any early detriment with APBI. If mature results of randomized trials confirm equivalence of APBI to conventional WBI in selected women, it would mark a paradigm shift and a major advance in treatment. This would allow many more women to opt for breast conservation, resolve the dilemmas regarding chemotherapy and radiotherapy sequencing and perhaps would be more cost effective. PMID- 16264856 TI - Therapy insight: Influence of type 2 diabetes on the development, treatment and outcomes of cancer. AB - Although type 2 diabetes and cancer are major health concerns among the adult population, few studies have directly addressed the relationship between the two, or the impact of diabetes on cancer outcomes. Diabetes and hyperglycemia are associated with an elevated risk of developing pancreatic, liver, colon, breast, and endometrial cancer. When treating cancer patients who have diabetes, clinicians must consider the cardiac, renal, and neurologic complications commonly associated with diabetes. Chemotherapeutic choices and, ultimately, the outcome for cancers may be affected by the avoidance of agents that have been shown to provide the best clinical response and survival in cancer patients without other disease complications. Evidence from population-based studies and clinical trials indicate that hyperglycemic and diabetic patients experience higher mortality and recurrence rates after diagnosis with, and treatment for, cancer. Evidence from the intensive care literature indicates that achieving glucose control leads to better clinical outcomes. If so, continued improvement of cancer outcomes may depend upon improved diabetes control. The association between diabetes and cancer is complex and warrants further study as the general population ages and the magnitude of both health problems continues to grow. Here we consider the influence of diabetes and hyperglycemia on the development, treatment, and long-term outcomes of cancer. PMID- 16264857 TI - Facial numbness in a man with inguinal and retroperitoneal masses. AB - BACKGROUND: A 37-year-old Brazilian man was admitted to Massachusetts General Hospital for evaluation of left-sided facial numbness, left-sided ataxia, dizziness, and vertigo. Seven weeks prior to admission, he reported numbness of the left oral cavity and tongue after a dental procedure. Three weeks prior to presentation, he developed left-sided incoordination with dizziness and vertigo. One week later, he noticed difficulty using a box cutter at work and presented to the emergency department for evaluation. A CT scan without contrast revealed no abnormalities and he was discharged home. Three days prior to admission, the patient developed diplopia. Cranial MRI revealed a 1.6 cm irregular enhancing mass in the left middle cerebellar peduncle. Neurologic examination was significant for diminished sensation over the left face to pinprick, left-sided dysmetria, and mild lateral instability of the trunk while walking. INVESTIGATIONS: MRI, lumbar puncture, CT scans of the abdomen and pelvis, needle biopsy of the lymph node, and paraneoplastic antibodies. DIAGNOSIS: Possible paraneoplastic demyelination. MANAGEMENT: Orchiectomy, adjuvant radiation, and corticosteroids. PMID- 16264858 TI - Treatment utopia. PMID- 16264859 TI - Importance of local control in an era of systemic therapy. PMID- 16264873 TI - Does docetaxel plus prednisone prolong the survival of men with metastatic hormone-refractory prostate cancer? PMID- 16264874 TI - Can docetaxel plus estramustine prolong survival of men with metastatic hormone refractory prostate cancer? PMID- 16264875 TI - Does the combination of rituximab and thalidomide influence the long-term perspectives of advanced-stage MCL? PMID- 16264876 TI - Should rituximab be used prior to autologous stem-cell transplantation for non Hodgkin's lymphoma? PMID- 16264877 TI - To irradiate or not to irradiate--that is the question. PMID- 16264878 TI - Is endoscopic ultrasonography superior to multidetector CT for assessing pancreatic cancer? PMID- 16264879 TI - Should paclitaxel be combined with epirubicin or carboplatin as first-line chemotherapy for advanced breast cancer? PMID- 16264880 TI - Therapy insight: Potential of statins for cancer chemoprevention and therapy. AB - The 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors, also known as statins, are commonly prescribed medications that lower serum cholesterol and decrease cardiac morbidity and mortality. These agents inhibit the rate-limiting step of the mevalonate pathway, an effect that influences cholesterol homeostasis and other diverse cellular functions. Preclinical data suggest statins have pleiotropic antineoplastic effects in a variety of tumors, but clinical studies have provided conflicting data regarding whether statins may increase or decrease the risk of cancer. Abnormal cholesterol metabolism in cancer is poorly understood but should be considered when evaluating the antineoplastic effects of statins. Emerging evidence suggests that atherosclerosis and cancer have similar underlying molecular mechanisms, both having lipid abnormalities and a pro-inflammatory phenotype. Like nonsteroidal anti-inflammatory agents, statins target lipid metabolism, have significant anti inflammatory effects, and can influence cardiovascular mortality. Recent studies show that statins may have chemopreventive effects and may complement cytotoxic chemotherapy or radiotherapy as a biologic response modifier in established cancer, but current data do not support their use as monotherapy. The preclinical data supporting anticancer activity, their additional health benefits, and the safety and relative low cost of statins compared to other 'targeted' agents currently under development all favor conducting prospective clinical trials of these drugs in cancer chemoprevention and therapy. PMID- 16264881 TI - Mechanisms of disease: Inflammation and the origins of cancer. AB - Many common cancers develop as a consequence of years of chronic inflammation. Increasing evidence indicates that the inflammation may result from persistent mucosal or epithelial cell colonization by microorganisms; including hepatitis B virus and hepatitis C virus, which can cause hepatocellular cancer; human papilloma virus subtypes, which cause cervical cancer, and the bacterium Helicobacter pylori, which can cause gastric cancer. At present, the cause of other chronic inflammatory conditions associated with increased cancer risk, such as ulcerative colitis, is obscure. Particular microbial characteristics as well as the type of the inflammatory response contribute to clinical outcomes via influence on epithelial cell and immune responses. Persistent inflammation leads to increased cellular turnover, especially in the epithelium, and provides selection pressure that result in the emergence of cells that are at high risk for malignant transformation. Cytokines, chemokines, free radicals, and growth factors modulate microbial populations that colonize the host. Thus, therapeutic opportunities exist to target the causative microbe, the consequent inflammatory mediator, or epithelial cell responses. Such measures could be of value to reduce cancer risk in inflammation-associated malignancies. PMID- 16264882 TI - Perspectives in the treatment of gastric cancer. AB - The overall 5-year survival of patients with gastric cancer is only 23% in the US compared with 60% in Japan. For Western patients, detecting the disease earlier and applying treatment quality control could substantially improve clinical outcome. For the treatment of gastric cancer, complete tumor resection, whenever feasible, is the standard treatment. Resection of the primary tumor (partial or total gastrectomy) is based on standardized criteria of the tumor, such as location, stage, histology, and surgical margins. The extent of regional lymphadenectomy required, however, has been a matter of considerable debate. Emerging evidence from the latest randomized controlled trials show that extended (D2) lymphadenectomy is safe and able to cure 20% of patients with N2-disease compared with 0% treated with limited D1 dissection, provided that the optimal surgical technique is used. Estimates suggest that this N2-specific subgroup advantage reflects a potential absolute overall survival benefit of 3-6%. Postoperative decisions about adjuvant chemotherapy and radiotherapy are based on pathologic staging, the extent of surgery performed (D0/D1 vs D2/D3) and the risk benefit ratio. Recurrence-risk and mortality-risk reduction is achievable with a carefully planned relapse-prevention guided therapeutic strategy. Patient-related factors (tumor features and expected recurrence-risk magnitude) and treatment related factors (surgical experience, adjuvant treatment risk-benefit ratio) should be considered on an individual basis. In future, genomic-based approaches will help to provide a more personalized therapeutic approach and improve patient outcome. PMID- 16264883 TI - Recent developments in therapeutic cancer vaccines. AB - Therapeutic cancer vaccines are being developed with the intention of treating existing tumors or preventing tumor recurrence. While the results of clinical trials, predominantly in the metastatic setting have been sobering, the central hypothesis of active immunotherapy i.e. that the human immune system can be activated to recognize and destroy tumor cells, remains a viable one. We believe that a fundamental shift in how clinical trials are performed, and what concepts they test is required to make meaningful strides towards future clinical use of cancer vaccines. First, we must reappraise whether the metastatic setting is the appropriate arena to test these agents. Second, we must arrive at a consensus on the most important biologic endpoints and rapidly test vaccines for their ability to achieve these endpoints. Third, we need to expend more effort on understanding how to manipulate the immune system beyond the initial stimulation provided by a vaccine. Fourth, in order to permit comparison of results across different studies, it would be helpful to narrow down the large number of vaccine platforms. We will discuss the current state of development of cancer vaccines and the relevance for future clinical use of these agents to treat and prevent cancers. PMID- 16264884 TI - Cancer immunotherapy comes of age. PMID- 16264885 TI - Breast pathology: rationale for adopting the ductal intraepithelial neoplasia (DIN) classification. PMID- 16264900 TI - What are the risk factors for brain metastasis in breast cancer patients? PMID- 16264901 TI - Is imatinib a cost-effective treatment for newly diagnosed chronic myeloid leukemia patients? PMID- 16264902 TI - How reliable is MRI for predicting extent of residual breast cancer with different primary medical therapies? PMID- 16264903 TI - Do we need to improve pain management in the radiation oncology department? PMID- 16264904 TI - Can molecular markers predict local, regional and distant metastasis in head and neck cancer? PMID- 16264905 TI - Is oral mucositis an inevitable consequence of intensive therapy for hematologic cancers? PMID- 16264906 TI - How should physicians communicate the transition to palliative care? PMID- 16264907 TI - Technology insight: Applications of emerging immunotherapeutic strategies for Epstein-Barr virus-associated malignancies. AB - The Epstein-Barr virus (EBV) is uniquely associated with a broad range of human malignancies. In spite of their diverse cellular origin, most of these malignancies share common features, including the expression of either some or all of the EBV latent proteins, which can be potentially exploited for immune based therapies. Here we discuss new and emerging strategies to manipulate the immune response to specifically boost T-cell immunity towards viral proteins that are expressed in EBV-associated malignancies. These strategies are used either alone or as an adjuvant therapy in combination with chemotherapy and/or monoclonal antibodies. Overall, this strategy may serve as a new paradigm for the successful multi-modality treatment of malignancies. PMID- 16264909 TI - Therapy insight: Cancer anorexia-cachexia syndrome--when all you can eat is yourself. AB - Tumor growth is associated with profound metabolic and neurochemical alterations, which can lead to the onset of anorexia-cachexia syndrome. Anorexia is defined as the loss of the desire to eat, while cachexia results from progressive wasting of skeletal muscle mass--and to a lesser extent adipose tissue--occurring even before weight loss becomes apparent. Cancer anorexia-cachexia syndrome is highly prevalent among cancer patients, has a large impact on morbidity and mortality, and impinges on patient quality of life. However, its clinical relevance is frequently overlooked, and treatments are usually only attempted during advanced stages of the disease. The pathogenic mechanisms of cachexia and anorexia are multifactorial, but cytokines and tumor-derived factors have a significant role, thereby representing a suitable therapeutic target. Energy expenditure in anorexia is frequently increased while energy intake is decreased, which further exacerbates the progressive deterioration of nutritional status. The optimal therapeutic approach to anorectic-cachectic cancer patients should be based on both changes in dietary habits, achieved via nutritional counseling; and drug therapy, aimed at interfering with cytokine expression or activity. Our improved understanding of the influence a tumor has on the host's metabolism is advancing new therapeutic approaches, which are likely to result in better preservation of nutritional status if started concurrently with specific antineoplastic treatment. PMID- 16264908 TI - Drug insight: Histone deacetylase inhibitors--development of the new targeted anticancer agent suberoylanilide hydroxamic acid. AB - This review focuses on the discovery and development of the histone deacetylase (HDAC) inhibitor, suberoylanilide hydroxamic acid (SAHA). Post-translational modifications of the histones of chromatin are important factors in regulating gene expression--so-called epigenetic gene regulation. Acetylation and deacetylation of lysine residues in histone tails, controlled by the activities of HDACs and histone acetyltransferases, are among the most studied post translational modification of histones. In addition to chromatin protein, transcription factors, cell-signaling regulatory proteins, and proteins regulating cell death are substrates of HDACs and may be altered in function by HDAC inhibitors. HDAC inhibitors have several remarkable aspects. For instance, despite HDACs being ubiquitously distributed through chromatin, SAHA selectively alters the transcription of relatively few genes, and normal cells are at least 10-fold more resistant than transformed cells to SAHA and related HDAC inhibitor induced cell death. HDAC inhibitors represent a relatively new group of targeted anticancer compounds, which are showing significant promise as agents with activity against a broad spectrum of neoplasms, at doses that are well tolerated by cancer patients. SAHA is one of the HDAC inhibitors most advanced in development. It is in phase I and II clinical trials for patients with both hematologic and solid tumors. PMID- 16264911 TI - The Framework Convention on Tobacco Control. PMID- 16264912 TI - The role of preoperative chemotherapy for esophageal cancer: when is surgery alone feasible? PMID- 16264910 TI - Multiple gastrointestinal stromal tumors and synchronous ileal carcinoids. AB - BACKGROUND: A 74-year-old African-American male presented with a 3-day history of hematemesis and melena. The patient reported no abdominal pain, constitutional symptoms, bright red blood per rectum, constipation, or diarrhea. His physical examination and medical history were unremarkable except for benign prostatic hypertrophy. An esophagogastroduodenoscopy showed a 3 x 2 x 2 cm smooth round mass in the cardia, 2 cm distal to the gastroesophageal junction. Biopsy of the mass revealed an ulcerated tumor composed of spindle cells. Immunohistochemistry showed positive staining for a number of biochemical markers, including KIT, Ki 67 and smooth muscle actin, but was negative for the markers S100 and desmin. A gastric-wedge resection revealed an ulcerated 4.5 cm mass in the stomach, and exploration of the abdomen revealed two ileal carcinoid tumors, jejunal diverticula and reactive mesenteric lymphadenopathy. INVESTIGATIONS: Esophagogastroduodenoscopy, biopsy, CT scan, immunohistochemistry, DNA microarray analysis and quantitative reverse transcriptase-PCR. DIAGNOSIS: Multiple gastrointestinal stromal tumors occurring concomitantly with ileal carcinoids. MANAGEMENT: Gastric-wedge resection and segmental resection. PMID- 16264926 TI - Re-induction of hormonal sensitivity in hormone-refractory prostate cancer--fact or fiction? PMID- 16264927 TI - What is the risk of second malignant neoplasms after childhood cancer? PMID- 16264928 TI - Is there a link between macronutrient intake and prostate cancer? PMID- 16264929 TI - Can combination salvage therapy for invasive aspergillosis improve patient outcome? PMID- 16264930 TI - Does secondary surgical cytoreduction improve survival in women with advanced ovarian cancer? PMID- 16264931 TI - Which should be added to cisplatin for advanced non-small-cell lung cancer- vinorelbine or gemcitabine? PMID- 16264932 TI - Can outpatient interleukin-2 and interferon replace high-dose interleukin-2 for metastatic renal cell carcinoma? PMID- 16264933 TI - Does addition of temozolomide to whole brain radiotherapy improve outcome in patients with brain metastases? PMID- 16264934 TI - Drug insight: New antiemetics in the management of chemotherapy-induced nausea and vomiting. AB - Nausea and vomiting remain among the most feared side effects of chemotherapy for cancer patients. Significant progress has been made in the last 15 years in developing more effective and better-tolerated measures to minimize chemotherapy induced nausea and vomiting (CINV). During the 1990s, the selective 5 hydroxytryptamine receptor antagonists were first introduced for the treatment of CINV, and resulted in more effective and better tolerated treatment of CINV. Despite recent progress, however, a significant number of patients still develop CINV, particularly during the 2-5 day period (delayed emesis) following chemotherapy. There is evidence that this may be an underappreciated problem on the part of some caregivers. Recently, two new antiemetics, aprepitant, the first member of the neurokinin-1 antagonists, and palonosetron, a second-generation 5 hydroxytryptamine receptor antagonist, received regulatory approval in the US. Both represent useful additions to the therapeutic armamentarium for the management of CINV. PMID- 16264936 TI - Intensity-modulated radiation therapy for the treatment of pediatric cancer patients. AB - Intensity-modulated radiation therapy (IMRT) is a novel form of radiotherapy, which has the potential to reduce the amount of radiation unintentionally delivered to normal tissues while maintaining a high radiation dose to the tumor in comparison with standard radiation techniques. In adults, this technology has been implemented in a number of tumor sites, but in children it has been little used. This article will review the current studies in which IMRT has been used in children. It will also discuss possible future applications for IMRT, and anticipated problems with its use. PMID- 16264937 TI - Palpable right breast mass in a pregnant woman. AB - BACKGROUND: A 29-year-old female presented with a palpable right breast mass at a 12-week prenatal visit. She had no family history of breast or ovarian cancer. Ultrasound revealed a 3 cm lobulated mass, which was confirmed to be malignant by a core biopsy. Postmastectomy pathology at 15 weeks' gestation demonstrated this mass to be a stage T2N0M0 high-grade invasive ductal carcinoma with 0/20 axillary nodes involved. A staging CT scan postpartum showed an enlarged right internal mammary lymph node, confirmed by MRI as suspicious for malignancy. INVESTIGATIONS: Physical examination, breast ultrasound, core biopsy, mastectomy, CT scan, MRI. DIAGNOSIS: Pregnancy-associated breast carcinoma. MANAGEMENT: Mastectomy, chemotherapy and radiotherapy. PMID- 16264935 TI - Mechanisms of disease: Inflammatory mediators and cancer prevention. AB - Discovery of molecular pathways critical to carcinogenesis is revolutionizing the treatment and prevention of cancer. Traditional chemotherapeutic approaches usually cause 'global' cytotoxicity to both normal and carcinoma cells. Over the past decade, however, investigators have developed compounds that inhibit tumor formation more selectively by targeting specific signaling pathways, including those involving the epidermal growth factor receptor (EGFR) and cyclooxygenase 2 (COX2). COX2-derived bioactive lipids, including prostaglandin E2, are potent inflammatory mediators that promote tumor growth and metastasis through stimulation of cell proliferation, invasion, and angiogenesis. Recent work has demonstrated significant crosstalk between the COX2 and EGFR pathways, while preclinical data demonstrates a synergistic effect when both pathways are targeted simultaneously. Combination therapy, a common strategy in cancer treatment, is likely to improve outcomes in cancer prevention as well. Ongoing clinical trials designed to assess whether low doses of COX2 and EGFR inhibitors used in combination could prove more effective and result in reduced toxicity than either agent alone may provide new options for cancer prevention and treatment. We discuss advances in cancer prevention by focusing on mechanisms by which bioactive lipids contribute to tumor formation. While cancer chemoprevention is a relatively young field, we argue that this approach to malignant disease bears significant potential. PMID- 16264939 TI - Bioinformatics in cancer therapeutics--hype or hope? PMID- 16264940 TI - Targeting the most critical cells: approaching leukemia therapy as a problem in stem cell biology. PMID- 16264953 TI - How does antiangiogenic therapy affect brain tumor response to radiation? PMID- 16264954 TI - Is fecal DNA testing superior to fecal occult-blood testing for colorectal cancer screening? PMID- 16264955 TI - Does androgen deprivation improve treatment outcomes in patients with low-risk and intermediate-risk prostate cancer? PMID- 16264956 TI - Chemotherapy for metastatic NSCLC: current status and future direction. PMID- 16264958 TI - How can bone turnover markers be best utilized for prediction of skeletal events in patients with solid tumors? PMID- 16264957 TI - Does childhood vaccination and exposure to infection improve long-term survival in patients with malignant melanoma? PMID- 16264959 TI - Is adjuvant hypofractionated radiotherapy appropriate for elderly patients with breast cancer? PMID- 16264960 TI - Technology insight: Application of molecular techniques to formalin-fixed paraffin-embedded tissues from breast cancer. AB - Breast cancer is a heterogenous disease in terms of both clinical behavior and molecular characteristics. To develop prognostic and predictive markers for breast cancer, it would be useful to be able to analyze formalin-fixed paraffin embedded tissue (FPET) collected and banked from completed clinical trials. RNAs extracted from FPETs are chemically modified and fragmented, and are therefore not ideal substrates for gene-expression profiling assays. However, methods are being developed to optimize the use of such RNAs for high-throughput gene expression profiling assays. For microarray analysis, existing methods may be adequate for fresh FPET, but they do not work well with older FPET. For older samples, real-time reverse transcription-polymerase chain reaction is the method of choice for gene-expression profiling. PMID- 16264961 TI - Mechanisms of disease: Prostate cancer--a model for cancer chemoprevention in clinical practice. AB - Prostate cancer is a significant cause of cancer death, making it an attractive target for chemoprevention. Epidemiologic studies and the Prostate Cancer Prevention Trial indicate that chemoprevention is possible. Strategies for prevention include hormonal manipulation and limiting accumulation of genetic damage with anti-inflammatory agents and/or dietary antioxidants. Development of an effective chemoprevention strategy for prostate cancer is evolving and will likely serve as a model for chemoprevention of other adult malignancies. PMID- 16264962 TI - Drug insight: thalidomide as a treatment for multiple myeloma. AB - Multiple myeloma (MM)--a malignancy of the bone marrow--remains incurable by current therapies, and there is an urgent need for new drugs based on a better understanding of the underlying disease biology. MM is characterized by monoclonal plasma cells that accumulate in the bone marrow, which provides a microenvironment that promotes tumor cell growth and survival and protection against various therapeutic agents. The MM cell interacts with bone marrow stromal cells and endothelial cells, as well as osteoblasts and osteoclasts. Our understanding of the tumor microenvironment has already prompted the development of new agents that are aimed at disrupting the multiple facets of these interactions. It has also enabled the development of a comprehensive and rational approach to preclinical evaluation of new agents, facilitating the translation of in vitro studies to in vivo tumor models and, subsequently, to clinical trials. In this review, we describe the preclinical studies that led to the development of clinical trials of thalidomide and its immunomodulatory derivatives as therapeutic agents for MM. These drugs, alone or in combination, have shown impressive activity at all stages of the disease, and these demonstrations of clinical benefit have in turn validated our model systems for drug discovery in MM. Integration of data from clinical trials and laboratory studies will allow the design of future clinical trials that combine thalidomide and its derivatives with other drugs, ultimately leading to more effective therapies and better outcomes in patients with MM. PMID- 16264963 TI - Multidisciplinary treatment of synchronous primary rectal and prostate cancers. AB - BACKGROUND: A 58-year-old Caucasian man with a history of irritable bowel syndrome and occasional rectal bleeding presented with a 4-week history of progressive, bright red blood per rectum. A digital rectal examination revealed a 3 cm distal, midrectal mass. Laboratory tests showed an elevated serum prostate specific antigen of 32 ng/ml but other physical and medical examinations were unremarkable. INVESTIGATIONS: Digital rectal examination, colonoscopy, rectal mass biopsy, endorectal ultrasound, transrectal ultrasound-guided prostate biopsy, CT scan and MRI. DIAGNOSIS: Clinical stage III (T3N1M0), moderately differentiated adenocarcinoma of the rectum and clinical stage II (T1cN0M0) adenocarcinoma of the prostate. MANAGEMENT: Intensity-modulated radiation therapy, chemoradiation, chemotherapy, hormone therapy and surgery. PMID- 16264964 TI - What phase III trials are needed to improve the treatment of advanced non-small cell lung cancer? PMID- 16264965 TI - Duration of adjuvant therapy for colorectal cancer: are we overtreating our patients? PMID- 16264980 TI - Does epidermal growth factor receptor status predict activity of cetuximab in colorectal cancer patients? PMID- 16264981 TI - Temporal trends in ovarian cancer: incidence and mortality across Europe. PMID- 16264982 TI - Nadir CA125 concentration as a prognostic indicator in ovarian cancer. PMID- 16264983 TI - What is the best way to manage patients treated with gefitinib for non-small-cell lung cancer? PMID- 16264984 TI - What are the prognostic factors for survival in patients with metastatic renal cell carcinoma? PMID- 16264985 TI - Does hematopoietic stem cell transplantation improve patient outcome in juvenile myelomonocytic leukemia? PMID- 16264986 TI - EGFR point mutation confers resistance to gefitinib in a patient with non-small cell lung cancer. PMID- 16264987 TI - Current challenges in Wilms' tumor management. AB - Wilms' tumor is a renal cancer that predominantly affects children during the first 2 years of life. The continuing success of clinical trials in Wilms' tumor patients over the past 30 years has led to an overall survival of 85%, and treatment-related morbidity has been reduced. Less-aggressive chemotherapeutic regimes are available for patients with validated good prognostic factors, such as low stage and favorable histology. It is becoming increasingly apparent that treatment can be optimized through stratification of patients according to tumor stage and histology. Established treatments for Wilms' tumor include perioperative vincristine and actinomycin, with or without doxorubicin or radiotherapy. Relapsed patients have the option of salvage chemotherapy with ifosfamide, carboplatin and etoposide, as well as high-dose chemotherapy regimes and autologous hemopoietic stem-cell rescue. Further research is required to refine these regimes and identify further the role of additional prognostic factors in this childhood disease. In this article we discuss the most-debated issues and advances that have been made in the management of Wilms' tumor. PMID- 16264988 TI - Alteration of radiotherapy fractionation and concurrent chemotherapy: a new frontier in head and neck oncology? AB - Despite recent advances in multimodality management of patients with stage III-IV head and neck squamous cell carcinoma, the prognosis in these patients remains disappointing. In an attempt to improve treatment outcome, several teams recently investigated the role of altered fractionation radiotherapy in conjunction with systemic chemotherapy. The controlled trials that investigated this combined approach indicate that, although the magnitude of its effect was less marked for survival indices than for local-regional control, the addition of chemotherapy to altered fractionation regimens results in a clear improvement for these endpoints compared with hyperfractionated or accelerated regimens alone. The key challenge now is to optimize the synergism of these regimens in order to increase their therapeutic ratio in terms of both local-regional and systemic outcomes. This review is a critical appraisal of the real opportunities offered by the application of treatments aimed at increasing the dose intensity of radiotherapy delivered concurrently with cytotoxic drugs. PMID- 16264989 TI - Mechanisms of disease: Insights into the emerging role of signal transducers and activators of transcription in cancer. AB - Members of the signal transducers and activators of transcription (STAT) pathway, which were originally identified as key components linking cytokine signals to transcriptional events in cells, have recently been demonstrated to have a major role in cancer. They are cytoplasmic proteins that form functional dimers with each other when activated by tyrosine phosphorylation. Activated STAT proteins translocate to the nucleus to regulate expression of genes by binding to specific elements within gene promoters. Constitutive activation of the STAT family members Stat3 and Stat5, and/or loss of Stat1 signaling, is found in a large group of diverse tumors. Increasing evidence demonstrates that STAT proteins can regulate many pathways important in oncogenesis including cell-cycle progression, apoptosis, tumor angiogenesis, tumor-cell invasion and metastasis, and tumor-cell evasion of the immune system. Based on these findings, a growing effort is underway to target STAT proteins directly and indirectly for cancer therapy. This review will highlight STAT signaling pathways, STAT target genes involved in cancer, evidence for STAT activation in human cancers, and therapeutic strategies to target STAT molecules for anticancer therapy. PMID- 16264990 TI - New concepts for phase I trials: evaluating new drugs combined with radiation therapy. AB - The rationale for delivering concomitant chemoradiation is not only to increase tumor cell kill but also to achieve a synergistic effect of chemotherapy and radiation. Combination of chemotherapy and radiotherapy has yielded encouraging results in patients with locally advanced diseases. Our increased knowledge of cancer at the molecular level has transformed our understanding of tumor radiation resistance. Preclinical models have shown that several biologic agents designed to target specifically these molecular processes are radiosensitizing agents. Many of these agents are in the process of clinical evaluation with radiotherapy. The translation of these findings into the clinical setting will be feasible only if early phase I trials demonstrate their safety when combined with ionizing radiation. The combination of new drugs and radiation might not necessarily be equivalent to the toxicity of the new drug plus the usual toxicity of radiation. The doses and schedule to be explored for the new drug might vary from those assessed for the new drug alone. Inappropriate evaluation of a combination regimen can result in unjustified abandonment of a combination, or adoption of a regimen at toxic dose levels because of poor toxicity monitoring. Beside the 'in field' radiation dose-dependent symptoms, 'outside the field' symptoms that are not dose dependent might be identified. Specific and long-term clinical evaluation will be required to identify potentially harmful interactions. It will be necessary to rethink phase I strategies, toxicity endpoints, and trial designs and concepts in order to fully optimize these regimens. PMID- 16264991 TI - Should the FDA be the doctor of last resort? PMID- 16264992 TI - Mortality and survival in breast and colorectal cancer. PMID- 16264993 TI - Phase III trials in oncology: setting standards of care? PMID- 16265007 TI - Do cyclo-oxygenase 2 inhibitors increase the risk of cardiovascular events? PMID- 16265008 TI - Can axillary radiotherapy replace axillary dissection for early-stage breast cancer? PMID- 16265009 TI - Is radiofrequency ablation effective in patients with early-stage hepatocellular carcinoma and cirrhosis? PMID- 16265010 TI - What are the long-term effects of chemotherapy and hormonal therapy for early breast cancer? PMID- 16265011 TI - Does postmastectomy radiation reduce mortality in women with stage T1-2 node positive breast cancer? PMID- 16265012 TI - Does treatment with recombinant human erythropoietin affect the survival of anemic patients with cancer? PMID- 16265013 TI - Can docetaxel dosing be individualized based on cytochrome p450 activity? PMID- 16265014 TI - Should sentinel lymph-node biopsy be used routinely for staging melanoma and breast cancers? AB - The sentinel lymph node (SLN) is the lymph node that represents the 'gate-keeper' of the lymphatic basin; it is the first node to receive lymphatic drainage from the site of the primary tumor. SLN biopsy is a staging procedure and should be considered as such; it is not meant to be a therapeutic operation. The SLN can be mapped and biopsied using tracer agents (e.g. radiolabelled colloid and/or vital blue dye), which are injected around the primary tumor site. Pathologic analysis of the SLN using a combination of serial sectioning of the node, standard hematoxylin and eosin staining, and immunohistochemistry decreases the false negative rate compared with traditional nodal processing. SLN biopsy is associated with lower morbidity than full lymphadenectomy. The SLN technique accurately reflects the metastatic status of the regional lymph-node basin; recurrent nodal disease in the mapped basin is rare following a tumor-free SLN biopsy result. The objectives of this review are to provide a current and concise overview of the current literature on SLN biopsy and describe its role in clinical oncology. PMID- 16265015 TI - Primer: an evidence-based approach to prognostic markers. AB - Prognostic markers can help to identify patients at different degrees of risk for specific outcomes, facilitate treatment choice, and aid patient counseling. Compared with other research designs, prognostic studies have been relatively neglected in the broad efforts to improve the quality of medical research, despite their ubiquity. Large protocol-driven, prospective studies are the ideal, with clear, unbiased reporting of the methods used and the results obtained. Unfortunately, published prognostic studies rarely meet such standards, and in this article we discuss their main problems and how they can be improved. In particular, an evidence-based approach to prognostic markers is required, as it is usually difficult to ascertain the benefit of a marker from single studies and a clear view is only likely to emerge from looking across multiple studies. Current systematic reviews and meta-analyses often fail to provide clear evidence based answers, and rather only draw attention to the paucity of good-quality evidence. Prospectively planned pooled analyses of high-quality studies, along with general availability of individual patient data and adherence to reporting guidelines, would help alleviate many of these problems. PMID- 16265016 TI - Long-term complete remission of laryngeal Kaposi's sarcoma after palliative radiotherapy. AB - BACKGROUND: A 43-year-old Caucasian man presented with enlarged right laterocervical lymph nodes, and a 6-month history of diarrhea and 9 kg weight loss. His previous medical history was unremarkable. Physical examination and a cervical CT scan showed several indistinct necrotic right laterocervical lymph nodes of 4 cm maximal diameter, but with no lesions of the respiratory or digestive tracts. INVESTIGATIONS: Physical examination, cervical CT scan, laterocervical lymph node biopsy, laboratory tests, and endoscopy. DIAGNOSIS: Clinical stage 4 AIDS with stage T0 I1 S1 epidemic AIDS-related Kaposi's sarcoma of the laterocervical lymph nodes and, subsequently, the larynx. MANAGEMENT: Highly active antiretroviral treatment (stavudine, didanosine, and nelfinavir), tracheotomy, and palliative radiotherapy. PMID- 16265018 TI - Gallstones: how do we translate an old story into future therapy? PMID- 16265019 TI - Treatment endpoints for advanced cholangiocarcinoma. PMID- 16265034 TI - What is the long-term effect of high-dose versus standard-dose omeprazole in patients with dyspepsia? PMID- 16265035 TI - Can paroxetine improve well-being in patients with irritable bowel syndrome who do not respond to a high-fiber diet? PMID- 16265036 TI - Does Helicobacter pylori infection and its eradication have any effect on heartburn and gastroesophageal reflux? PMID- 16265037 TI - What is the most accurate test to differentiate pancreatic cystic neoplasms? PMID- 16265038 TI - Are covered stents superior to uncovered stents in the management of malignant biliary obstruction? PMID- 16265039 TI - Is mortality from liver disease associated with normal serum aminotransferase concentrations? PMID- 16265040 TI - Can epoetin alfa maintain ribavirin doses in patients with hepatitis C virus on combination therapy? PMID- 16265041 TI - Update on the etiology, pathogenesis and diagnosis of ulcerative colitis. AB - Evidence is accumulating that both genetic and environmental factors contribute to ulcerative colitis. The most consistent genetic associations have been shown for the MHC locus HLA Class II alleles, but the interleukin-1 family of genes and the multidrug resistance gene MDR1 have also been implicated as genetic susceptibility factors for the development of disease. In addition, there is a relationship between ulcerative colitis and bacterial flora, with an increased number of adherent Bacteroides spp. and Enterobacteriaceae spp. present in inflamed bowel segments. Conversely, cigarette smoking and appendectomy have both been shown to protect against the development of ulcerative colitis. Despite our improved understanding of the genetics and inflammatory mechanisms that underpin this disease, however, the etiology and pathogenesis of ulcerative colitis remain undefined. The diagnosis of ulcerative colitis is being aided by recent advances in diagnostic strategies, including the detection of fecal and serologic markers and the use of wireless capsule endoscopy, but, in the absence of a pathognomonic marker, the definition of this disease remains based on well-established clinical, endoscopic and histologic criteria. In particular, it is difficult to discriminate ulcerative colitis from other forms of colitis, including Crohn's disease, and there seems to be a growing overlap of pathophysiologic processes between ulcerative colitis and post-infectious irritable bowel syndrome. Patients who remain indeterminate between ulcerative colitis and Crohn's disease also continue to be a diagnostic challenge. PMID- 16265042 TI - Treatment of recurrent Clostridium difficile-associated disease. AB - Recurrent Clostridium difficile-associated disease (RCDAD) is a difficult treatment problem--once a patient has one recurrence of the disease the likelihood of further recurrences is markedly increased. Repeat antibiotics are usually indicated, either metronidazole or vancomycin. Tapering and pulsing the antibiotic dose after a 10-day standard course decreases the incidence of recurrences compared with abruptly stopping antibiotics after a simple 10-day course. If recurrences continue after two courses of metronidazole, vancomycin may be preferable to avoid the risk of neurotoxicity that is associated with prolonged metronidazole use. There is also a role for probiotics in the treatment of RCDAD; Saccharomyces boulardii has been shown to decrease recurrences by about 50%, especially when combined with high-dose vancomycin. Other microbiologic approaches include the restoration of normal colonic flora by fecal enema or by nasogastric tube, but these have not been studied in controlled trials. In addition, there are numerous new treatment approaches that are under development, including a vaccine, which promise to aid the future treatment of RCDAD as well as prevention of initial CDAD. PMID- 16265043 TI - Mechanisms of disease: The role of hepcidin in iron homeostasis--implications for hemochromatosis and other disorders. AB - The defensin-like circulatory peptide hepcidin is the iron-regulatory hormone that links innate immunity and iron metabolism. In response to inflammatory stimuli, the liver produces hepcidin: this antimicrobial peptide then limits the iron that is vital to invading pathogens, by decreasing iron release/transfer from enterocytes and macrophages and causing secondary hypoferremia. This may lead, however, to reduced iron availability for erythropoiesis and therefore to anemia (and anemia of chronic disease). When iron is scarce, the rate at which it is released into the bloodstream must be enhanced: indeed, iron starvation and hypoxia readily abrogate hepcidin expression. Conversely, if excess iron enters the circulation, hepcidin transcription is turned on and iron release from the intestine and macrophages abrogated. Circumstantial evidence indicates that the effect of circulatory iron on hepcidin requires functional HFE and hemojuvelin, two proteins of unknown function that have recently been linked to human hereditary hemochromatosis. In this disease it is likely that inadequate levels of circulating hepcidin lead to the uncontrolled release of iron from the intestine and macrophages, followed by tissue iron overload and organ damage. Given its role as the iron-regulatory hormone, the modulation of hepcidin activity using agonists or antagonists might offer new treatment opportunities in different human iron-dependent disorders. PMID- 16265044 TI - Mechanisms of disease: Advances in understanding the mechanisms leading to chronic pancreatitis. AB - Chronic pancreatitis remains a challenging and frustrating clinical problem. In the past few years, however, advances in genetic and immunologic research have spawned new insights and approaches to chronic pancreatitis. Genetic and environmental risk assessment may help identify individuals who are likely to develop severe chronic pancreatitis early in the disease course, and allow targeted attention to reduce confounding risks and slow or prevent this problem in the future. PMID- 16265045 TI - Cholestatic jaundice and bone lesions in an elderly woman. AB - BACKGROUND: A 74-year-old Caucasian female presented with jaundice, clay-colored stools, diarrhea, and fatigue of 3 months' duration, accompanied by a weight loss of 6.8 kg. Physical examination demonstrated mild hepatomegaly. Initial blood work revealed abnormal liver biochemistries with a cholestatic pattern. An abdominal CT scan showed intrahepatic bile-duct dilatation without masses but with multiple lytic and blastic bone lesions. A sacral bone biopsy established the diagnosis. INVESTIGATIONS: Endoscopic ultrasound, endoscopic retrograde cholangiopancreatography, CT scan, bone biopsy and liver biopsy. DIAGNOSIS: Systemic mastocytosis affecting the biliary system resulting in a primary sclerosing-cholangitis-like picture combined with diffuse blastic and lytic bone lesions. MANAGEMENT: Biliary stenting, histamine 1- and 2-receptor blockers, and chemotherapy (cladribine). PMID- 16265046 TI - Should you "just say 'no'"? PMID- 16265047 TI - Is the use of COX-2 inhibitors in gastroenterology cost-effective? PMID- 16265048 TI - Can NOD2/CARD15 mutations predict intestinal graft-versus-host disease and aid our understanding of Crohn's disease? PMID- 16265063 TI - Is the current standard therapy for hepatitis C of peginterferon and ribavirin an option for patients coinfected with HIV? PMID- 16265064 TI - Can the addition of bevacizumab to IFL chemotherapy improve outcome in colorectal cancer? PMID- 16265065 TI - Are miniature ultrasonic probes of clinical value for diagnosis and treatment of digestive tract diseases? PMID- 16265066 TI - Long-term outcome improvement in patients with chronic hepatitis D treated with interferon alpha. PMID- 16265067 TI - Is transarterial chemoembolization an option for all patients with unresectable hepatocellular carcinoma? PMID- 16265068 TI - Maintenance therapy versus episodic therapy with infliximab for Crohn's disease. PMID- 16265069 TI - Treating chronic anal fissure with botulinum neurotoxin. AB - Recent reports confirm that the management of chronic anal fissure has undergone extensive re-evaluation during the past few years. This rejuvenation of interest is attributable to the application of neurochemical treatment, which has contributed to the tendency to treat the disease on an outpatient basis. The use of botulinum neurotoxin seems to be a promising and safe approach for the treatment of chronic anal fissure, particularly in patients at high risk for incontinence. Indeed, botulinum neurotoxin has been successfully used selectively to weaken the internal anal sphincter as a treatment for chronic anal fissure. It is also more efficacious than nitrate therapy, and is not related to the patient's willingness to complete treatment. PMID- 16265070 TI - Drug insight: Nucleoside and nucleotide analog inhibitors for hepatitis B. AB - In the past decade, substantial advances have been made in the treatment of chronic hepatitis B. Approved treatments include interferon-alpha, the nucleoside analog inhibitor lamivudine, and the nucleotide analog inhibitor adefovir dipivoxil. This review provides insights into the benefits and limitations of lamivudine and adefovir dipivoxil for the treatment of chronic hepatitis B. Lamivudine and adefovir dipivoxil have similar antiviral efficacies. Lamivudine has negligible side effects but a high rate of drug resistance, whereas adefovir dipivoxil has a low rate of drug resistance but long-term use is associated with a small risk of nephrotoxicity. Several other nucleoside and nucleotide analogs are being evaluated in phase II/III clinical trials for hepatitis B. PMID- 16265071 TI - Mechanisms of disease: Mechanisms of hepatic fibrosis and therapeutic implications. AB - Hepatic fibrosis, or scarring of the liver, is emerging as a treatable complication of advanced liver disease, following significant progress in understanding its underlying mechanisms. Efforts have focused on the hepatic stellate cell, as these cells can undergo 'activation' into proliferative and fibrogenic myofibroblast-like cells during liver injury. Stimuli driving stellate cell activation include hepatocellular necrosis due to oxidant stress, apoptosis, and soluble growth factors. Specific lymphocyte subsets can also stimulate fibrogenesis. A cascade of signaling and transcriptional events in stellate cells underlies the fibrogenic response to liver injury, with each step in the cascade being a potential target for antifibrotic therapy. Disease-specific fibrogenic mechanisms have also been uncovered: in hepatitis C, this may include direct stimulation of stellate cell activation by viral infection; in nonalcoholic steatohepatitis, elevated levels of leptin and increased leptin signaling by stellate cells increase fibrogenesis. Determinants of fibrosis progression include both environmental and genetic factors, with ongoing efforts to define specific polymorphisms correlating with fibrosis progression rates. Human studies now indicate that fibrosis and even cirrhosis could be reversible, especially if the underlying disease is eradicated. A key challenge is to establish noninvasive means of assessing fibrosis stage and progression using either serum tests and/or imaging. In addition, endpoints of antifibrotic clinical trials need to be established so that reliable evidence of benefit can be identified. We are on the cusp of a new era in which antifibrotic therapies could become important in treating chronic fibrosing liver disease. PMID- 16265072 TI - Mechanisms of disease: Carcinogenesis in Barrett's esophagus. AB - The pathogenesis of cancer in Barrett's esophagus is multifactorial. Gastroesophageal reflux seems to be important in the initiation of Barrett's esophagus, but its role in promoting carcinogenesis has yet to be established. Diet, lifestyle and carcinogens, especially the nitrates, may be important in the development of carcinogenesis, and require further investigation. Inhibition of reflux-stimulated inflammatory changes, for example by inhibiting cyclooxygenase, holds promise for decreasing cancer progression. Similarly, dietary and lifestyle modification used in the management of reflux may also help to prevent the development of esophageal cancer. The molecular changes that are associated with the development of cancer in Barrett's esophagus offer several potential areas of intervention to prevent and manage esophageal cancer. Limiting cell growth, increasing apoptosis of damaged cells, limiting cell invasion and angiogenesis factors could be useful to accomplish this goal. Having a greater understanding of the pathogenesis of this condition can only help to develop more management options in the future. PMID- 16265073 TI - Fever and right upper quadrant pain in a pregnant woman at 30 weeks' gestation. AB - BACKGROUND: A 34-year-old gravid woman with a history of ileal Crohn's disease presented at 30 weeks' gestation with a 2-week history of fever and right upper quadrant pain. An intra-abdominal abscess was suspected. Ultrasound and MRI failed to demonstrate the suspected abscess. Owing to ongoing pain and fever, the risk to the fetus of a CT scan were discussed with the patient, obstetricians and radiologists, with considerable debate about the possibility of other explanations for her symptoms. Ultimately, a CT scan revealed marked thickening of the distal ileum and confirmed diagnosis of an abscess in continuity with the inflamed bowel. INVESTIGATIONS: Ultrasound, MRI, CT scan, urinalysis, urine culture and liver function tests. DIAGNOSIS: Crohn's disease flare complicated by an intra-abdominal abscess. MANAGEMENT: Antibiotics (ceftizoxime, metronidazole and amoxicillin/clavulanate potassium), parenteral nutrition and ileocecectomy. PMID- 16265074 TI - Conflicts of interest. PMID- 16265075 TI - Are laparoscopic antireflux procedures useful? PMID- 16265076 TI - Are endoscopic antireflux procedures useful? PMID- 16265092 TI - Safety and efficacy of lumiracoxib compared with NSAIDs. PMID- 16265091 TI - Does the addition of lamivudine to peginterferon alpha-2a sustain response rates in HBeAg-negative hepatitis B? PMID- 16265093 TI - Is ultrasound-guided fine-needle biopsy effective for diagnosis of early HCC in liver cirrhosis? PMID- 16265094 TI - Is treatment with nadolol effective against the growth of small esophageal varices in patients with cirrhosis? PMID- 16265095 TI - Can the addition of cetuximab to irinotecan improve outcome in colorectal cancer? PMID- 16265096 TI - Value of screening the laryngopharyngeal area during routine upper gastrointestinal endoscopy. PMID- 16265097 TI - Patient selection in an era of donor liver shortage: current US policy. AB - In the past, organ allocation in the US was based on anecdotal experience, self interest and the opinions of single centers, with little support in the way of scientific evidence, mathematical survival modeling or validation. As organ transplantation became more successful, and as disparity between the number of patients on the waiting list and available organs became larger, a more justifiable donor allocation scheme became necessary. The current allocation scheme for donor livers is based on the model for end-stage liver disease/pediatric end-stage liver disease, which was introduced in 2002 by the United Network for Organ Sharing. This new allocation system has improved accuracy for predicting pretransplant mortality. In addition, the number of liver transplantations has risen for almost all etiologic categories, most noticeably for patients with hepatocellular carcinoma. Fewer patients have been registered on the liver transplant waiting list and fewer have been removed from the list because they have died or become too sick for transplantation. So far, this new allocation system has been a success, but it does have its shortcomings, and even with improvements to the system, the use of the donor organ pool still needs to be optimized. PMID- 16265098 TI - Technology insight: Laser-scanning confocal microscopy and endocytoscopy for cellular observation of the gastrointestinal tract. AB - Recent advances in endoscopic imaging technology have enabled the visualization of early-stage cancer and its precursors in the gastrointestinal tract. Chromoendoscopy, magnifying endoscopy, endoscopic optical coherent tomography, spectroscopy, and various combinations of these technologies, are all important for the recognition of small and unclear lesions. To observe cancer cells in vivo, two types of ultra-high magnifying endoscope--'laser-scanning confocal endoscopy series' and 'contact endoscopy series'--that have a maximum of more than 1,000x magnifying power have been developed. These endoscopes can generate high-quality images of both living cancer cells and normal cells in the gastrointestinal tract, with a quality comparable to that possible with conventional cytology. These novel imaging technologies may make in vivo histological diagnosis by virtual histology possible. PMID- 16265099 TI - Technology insight: Current status of endoscopic ultrasonography. AB - Endoscopic ultrasound (EUS) has become the most accurate imaging modality for locoregional cancer staging of the gastrointestinal tract. Fine-needle aspiration (FNA) has added a new level of accuracy for EUS in nodal staging, with reported numbers in the 90% range for luminal and pancreaticobiliary disease. In addition, new non-gastrointestinal applications are being evaluated, such as the role of EUS-FNA for the staging of non-small-cell lung cancer and exploration of the posterior mediastinum. Furthermore, the same techniques that make safe tissue sampling possible are being explored for their use as interventional applications, such as EUS-guided celiac plexus neurolysis, fine-needle injection, EUS-guided pseudocyst drainage, and EUS-guided cholangiography and pancreatography. This review describes the current clinical status of EUS in gastrointestinal oncology, as well as future and novel indications and therapeutic strategies for this technology. PMID- 16265100 TI - Mechanisms of Disease: pathogenesis of nonalcoholic fatty liver disease. AB - Nonalcoholic fatty liver disease (NAFLD) is associated with the metabolic syndrome. The metabolic syndrome is characterized by insulin resistance, which is produced by a complex interaction between genetic factors, macronutrient intake and lifestyle that alters the cytokine profile, cell biology and biochemical milieu of the liver, adipose tissue and striated muscle. The resultant disequilibrium in lipid homeostasis causes triglycerides to accumulate in the liver. An increase in oxidative stress, due to the generation of reactive oxygen species as a result of mitochondrial abnormalities and induction of the cytochrome P-450 system could be one mechanism by which the nonalcoholic fatty liver develops into nonalcoholic steatohepatitis. The pathogenesis of cytologic ballooning and Mallory body formation and their role in NAFLD remain to be defined. In addition, inflammation and fibrosis are likely to be secondary to hepatocyte injury and death. PMID- 16265101 TI - Percutaneous treatment of a patient with infected necrotizing pancreatitis. AB - BACKGROUND: A 50-year-old man with an acute episode of alcohol-induced chronic edematous pancreatitis and an inflammatory stenosis of the biliary tract was treated by implantation of a plastic endoprosthesis at a rural hospital in Germany. Because of his worsening condition, the patient was referred to the intensive care unit of the University Hospital, Regensburg, Germany. Contrast enhanced CT revealed complete necrosis of the body and tail of the pancreas, formation of large retrogastric and paraduodenal fluid collections, and fluid along Gerota's fascia, as well as in the paracolic gutters. Antibiotic treatment was adjusted according to the results of microbiological testing after diagnostic puncture of the necrotic cavity. INVESTIGATIONS: CT scan, CT-guided fine-needle aspiration and fluoroscopy. DIAGNOSIS: Infected acute necrotizing pancreatitis. MANAGEMENT: Interventional treatment using large-bore percutaneous catheters to perform percutaneous necrosectomy, fragmentation of necrotic pancreatic tissue with a snare catheter and dormia basket, and aspiration. Parenteral nutrition and antibiotics were also administered. PMID- 16265102 TI - The whole is greater than the sum. PMID- 16265103 TI - The value of surveillance and other unresolved issues in the management of Barrett's esophagus. PMID- 16265104 TI - Is there a role for helminths in the therapy of inflammatory bowel disease? PMID- 16265119 TI - Helicobacter pylori test-and-eradicate strategy versus early endoscopy in patients with dyspepsia. PMID- 16265120 TI - Risk of community-acquired pneumonia after acid-suppressive drugs. PMID- 16265121 TI - Should endoscopic balloon dilation be used for the extraction of bile duct stones? PMID- 16265122 TI - Low dose versus high dose imatinib for gastrointestinal stromal tumors. PMID- 16265123 TI - Safety and efficacy of endoscopic mucosal resection in patients with lateral spreading tumors of the colorectum. PMID- 16265124 TI - When should chemoradiotherapy for rectal cancer be done? PMID- 16265125 TI - Treatments targeting putative mechanisms in irritable bowel syndrome. AB - The pathophysiology of irritable bowel syndrome (IBS) is heterogeneous; it is possible for several mechanisms to be disturbed in the same patient. Isolating a single target for pharmacological manipulation is also difficult because of the complexity and overlap of the neural circuitry in the enteric and central nervous system. This review summarizes the rationale and efficacy of current and future therapies for IBS, on the basis of putative pathophysiological models. The modulation of gastrointestinal sensorimotor function, intestinal gas handling, the gastrocolonic reflex, neurohormonal stress responses, central processing of afferent information, and microbial flora are the current frontiers for experimental therapeutics for IBS. Patients presumed to have POSTINFECTIOUS IBS have also been targeted as a distinct group. In the very near future, it is unlikely that a single drug will come to the fore as a suitable and successful treatment for everyone with IBS, but new data on potential therapeutic targets lend hope for the improved long-term management of IBS. Disease modification rather than just symptom-based treatments must remain the goal. PMID- 16265126 TI - Drug Insight: appetite suppressants. AB - The term 'appetite suppressant' is used to denote drugs that act primarily on the neurochemical transmitters of the central nervous system to reduce food intake. In addition to drugs that release or mimic the effect of norepinephrine (noradrenaline), this could include drugs that inhibit: reuptake of norepinephrine or 5-hydroxytryptamine (also known as serotonin); bind to the gamma-aminobutyric acid receptors or the cannabinoid receptors; and some peptides that reduce food intake. The sympathomimetic drugs phentermine, diethylpropion, benzphetamine, and phendimetrazine--so named because they mimic many effects of norepinephrine--are only approved in a few countries, and then only for short term use. Sibutramine, a norepinephrine-5-hydroxytryptamine reuptake inhibitor, is approved for long-term use. Several new mechanisms for drug action are under investigation. Appetite suppressants should be viewed as useful adjuncts to diet and physical activity and might help selected patients to achieve and maintain weight loss. PMID- 16265127 TI - Technology insight: calprotectin, lactoferrin and nitric oxide as novel markers of inflammatory bowel disease. AB - Distinguishing patients with inflammatory bowel disease from those with irritable bowel syndrome can be difficult. A simple and reliable test that detects intestinal inflammation would therefore be very useful in the clinic. If such a test parameter correlated with the intensity of the inflammatory reaction it could also be used to monitor disease activity. Calprotectin, lactoferrin and nitric oxide are produced and released locally in much greater quantities in the inflamed gut than in the noninflamed gut. These compounds can be readily measured in fecal samples (calprotectin and lactoferrin) or directly in the intestinal lumen (nitric oxide gas). Here, we discuss what is known about these markers, how they could be used in clinical practice and how they can complement existing techniques used for the diagnosis and monitoring of inflammatory bowel disease. PMID- 16265128 TI - Mechanisms of disease: adipocytokines and visceral adipose tissue--emerging role in intestinal and mesenteric diseases. AB - Adipose tissue has long been regarded as a passive type of connective tissue that stores energy as triglycerides and releases energy as free fatty acids, however, this point of view has now changed. The wide variety of products expressed and secreted by adipose tissue, such as adiponectin, leptin, and resistin, mean that the total adipose tissue mass can be defined as a real endocrine organ. The anatomic, metabolic and biochemical characteristics of visceral adipose tissue make it interesting in the context of intestinal and mesenteric diseases. These characteristics include increased lipolysis, venous drainage via the portal vein and local glucocorticoid excess owing to the specific expression of 11-beta hydroxysteroid-dehydrogenase type 1. In this review, the role of the visceral adipose tissue and its secretory products in intestinal and mesenteric diseases is systematically reviewed, with special focus on 'creeping fat' in Crohn's disease and mesenteric panniculitis. PMID- 16265130 TI - Case studies--the appliance of science. PMID- 16265129 TI - Hematological malignancy manifesting as ascites. AB - BACKGROUND: A 63-year-old Caucasian woman presented with intermittent, left-sided abdominal discomfort without other symptoms. Physical examination revealed massive splenomegaly and complete blood counts showed thrombocytopenia. Splenectomy was recommended but the patient declined. She was lost to follow-up twice but returned with tense ascites about 2 years after the initial presentation. Despite aggressive medical management, the ascites did not improve. INVESTIGATIONS: Bone-marrow and liver biopsies, abdominal ultrasound, esophagogastroduodenoscopy, abdominal CT scan and peripheral blood smear. DIAGNOSIS: Primary splenic lymphoma with hepatic infiltration causing portal hypertension and ascites. MANAGEMENT: Paracentesis, dietary sodium restriction and diuretics, splenectomy, splenorenal shunt and chemotherapy (cyclophosphamide, doxorubicin, vincristine and prednisone). PMID- 16265131 TI - Molecular genetics in gastroenterology: from research success to clinical application? PMID- 16265132 TI - Functional heartburn: misdiagnosed gastroesophageal reflux or a distinct entity? PMID- 16265150 TI - Can Peg-IFN alpha-2a plus ribavirin be used to treat patients with chronic hepatitis C and normal alanine aminotransferase levels? PMID- 16265151 TI - Can serum markers be used to reliably detect liver fibrosis? PMID- 16265152 TI - What is the most sensitive screening method for the detection of colon cancer? PMID- 16265153 TI - Photodynamic therapy for the treatment of patients with Barrett's high-grade dysplasia and mucosal adenocarcinoma. PMID- 16265154 TI - Is lamivudine a safe and effective therapy for patients with chronic hepatitis B and advanced liver disease? PMID- 16265155 TI - Future therapeutic options for celiac disease. AB - Celiac disease is a disorder of the small intestine caused by an inappropriate immune response to wheat gluten and similar proteins of barley and rye. At present, the only available treatment is a strict gluten-exclusion diet; hence the need for alternative treatments. Recent advances have improved our understanding of the molecular basis for this disorder and there are several attractive targets for new treatments. Oral enzyme supplementation is designed to accelerate gastrointestinal degradation of proline-rich gluten, especially its proteolytically stable antigenic peptides. Complementary strategies aiming to interfere with activation of gluten-reactive T cells include the inhibition of intestinal tissue transglutaminase activity to prevent selective deamidation of gluten peptides, and blocking the binding of gluten peptides to the HLA-DQ2 or HLA-DQ8 molecules. Other possible treatments include cytokine therapy, and selective adhesion molecule inhibitors that interfere with inflammatory reactions, some of which are already showing promise in the clinic for other gastrointestinal diseases. PMID- 16265157 TI - Bleeding gastric varices and antiphospholipid syndrome. AB - BACKGROUND: A 35-year-old woman with a past history of antiphospholipid syndrome and near-fatal vascular thrombosis was transferred to Glasgow Royal Infirmary, UK, for investigation of 'obscure' gastrointestinal bleeding in the context of long-term oral anticoagulation therapy. Initial endoscopic examination revealed isolated gastric varices with stigmata of recent hemorrhage. She had no prior history of liver disease or portal hypertension. Investigations to ascertain the cause of her varices were performed. INVESTIGATIONS: CT venography, endoscopic ultrasound and transjugular liver biopsy. DIAGNOSIS: Nodular regenerative hyperplasia of the liver leading to gastric varices. MANAGEMENT: Endoscopic histacryl tissue-glue injection followed by transjugular intrahepatic portosystemic shunt and long-term oral anticoagulation therapy. PMID- 16265160 TI - The 'upside' of travelers' diarrhea: a personal case report. PMID- 16265156 TI - Treatment strategies in nonalcoholic fatty liver disease. AB - Nonalcoholic fatty liver disease (NAFLD) is an increasingly recognized health problem. Increased fat accumulation in the liver is observed in 20-30% of the population in the Western world, and in approximately 10% of this cohort it is associated with nonalcoholic steatohepatitis, which is characterized by inflammation and fibrosis. Disease presentation of NAFLD ranges from asymptomatic disease to cirrhosis with the complication of liver failure and hepatocellular carcinoma. NAFLD is suspected on the basis of various clinical aspects (an elevated alanine aminotransferase concentration, presence of obesity and diabetes) that alone are not sufficient to establish diagnosis or prognosis. The major diagnostic procedure is liver biopsy, which allows assessment of liver injury. In most cases, NAFLD is associated with insulin resistance, which is therefore the target of most current NAFLD treatment modalities. Various treatment strategies such as weight loss and/or exercise, thiazolidinediones, metformin, lipid-lowering agents and antioxidants have been studied. So far, no single intervention has convincingly improved liver histology. It is recommended that patients at high risk of developing advanced liver disease, and who are not part of controlled studies, should receive nutritional counseling and take physical exercise to achieve moderate weight loss and improve insulin sensitivity. PMID- 16265177 TI - Is magnetic resonance cholangiography of clinical value in the diagnosis and treatment of gallstone pancreatitis? PMID- 16265178 TI - Can OCT be used to distinguish accurately between patients with ulcerative colitis and those with Crohn's disease? PMID- 16265179 TI - How effective are quadruple therapies as first-line H. pylori eradication therapies? PMID- 16265180 TI - Is endoscopic ultrasonography superior to multidetector CT for assessing pancreatic cancer? PMID- 16265181 TI - Can the risk of pancreatitis following endoscopic papillary balloon dilation for common bile duct stones be reduced? PMID- 16265182 TI - How effective are rescue H. pylori eradication therapies in patients with dyspepsia? PMID- 16265183 TI - Update on the classification, assessment of prognosis and therapy of Budd-Chiari syndrome. AB - Budd-Chiari syndrome (BCS) occurs as a result of obstruction of hepatic venous outflow at any level from the small hepatic veins to the junction of the inferior vena cava with the right atrium. Diagnosis can be difficult because of the wide spectrum of presentation of the disease and the varying severity of liver damage. The traditional classification of BCS--as fulminant, acute or chronic--is not prognostically useful. This makes assessing the benefit of therapy difficult, especially as there is no evidence from randomized studies. This article highlights advances in the prognosis and therapy of BCS. Identification of the site of venous obstruction has a major effect on prognosis. Portal-vein thrombosis occurs in 20-30% of cases, and acute presentation of BCS reflects an acute or chronic syndrome in 60% of BCS cases. BCS can be diagnosed and treated on a single occasion in the setting of the radiology department, with hepatic venography, transjugular liver biopsy, retrograde CO2 portography and inferior vena cava pressure measurements performed simultaneously with therapies such as dilation or stenting of webs in the inferior vena cava or hepatic veins, and placement of transjugular intrahepatic portosystemic shunts. Disruption of a portal vein thrombus can also be done during the same session. Surgical shunts have been superseded by the use of transjugular intrahepatic portosystemic shunts. Liver transplantation is reserved for fulminant and progressive chronic forms of BCS. Anticoagulation therapy must be used routinely, before and after specific therapy, regardless of whether a thrombophilic disorder is diagnosed. PMID- 16265184 TI - Travelers' diarrhea: antimicrobial therapy and chemoprevention. AB - The use of preventive measures and self-treatment for travelers' diarrhea is routine in regions where the occurrence of diarrhea is predictably high. People traveling to these areas who do not exercise care in their selection of consumed foods and beverages will suffer high rates of illness. Such diarrhea normally affects the traveler for a day, although it can result in chronic postinfectious irritable bowel syndrome. Although systemic antibacterial drugs are effective in preventing diarrhea, their use is not routinely recommended because of side effects and their importance as a therapy for extra-intestinal infections. This review focuses on current and future uses of antibacterial drugs in the prevention and therapy of travelers' diarrhea. Minimally absorbed (< 0.4%) rifaximin can effectively reduce the occurrence of travelers' diarrhea without side effects. Bismuth subsalicylate is a useful alternative, although it is less effective than rifaximin for the prevention of travelers' diarrhea and the required doses are less convenient. All people who travel to high-risk areas should take curative antimicrobial agents with them for self-treatment of illness: rifaximin 200 mg three times a day for 3 days, or an absorbable agent such as a fluoroquinolone or azithromycin taken in a single dose initially, with the need for a second or third dose determined by clinical response. Loperamide (up to 8 mg per day for < or = 2 days) can be given with the antibiotic to offer rapid symptomatic improvement. In the future, the ability to evaluate the genetic risk of illness acquisition might allow person-specific recommendations to be made. PMID- 16265185 TI - Assessment, prevention and treatment of osteoporosis in inflammatory bowel disease patients: a bone of contention. PMID- 16265186 TI - Is there a role for apheresis in gastrointestinal disorders? PMID- 16265200 TI - Should nonalcoholic fatty liver disease be treated differently in elderly patients? PMID- 16265201 TI - Effectiveness of controlled-release budesonide in preserving bone mineral density in patients with Crohn's disease. PMID- 16265202 TI - What are the effects of cyclooxygenase-2-specific inhibitors on the small bowel? PMID- 16265203 TI - How useful is contrast-enhanced ultrasonography in the diagnosis of intraductal papillary mucinous tumors? PMID- 16265204 TI - Diarrhea caused by primarily non-gastrointestinal infections. AB - Infectious diseases that do not primarily affect the gastrointestinal tract can cause severe diarrhea. The pathogenesis of this kind of diarrhea includes cytokine action, intestinal inflammation, sequestration of red blood cells, apoptosis and increased permeability of endothelial cells in the gut microvasculature, and direct invasion of gut epithelial cells by various infectious agents. Of the travel-associated systemic infections presenting with fever, diarrhea occurs in patients with malaria, dengue fever and SARS. Diarrhea also occurs in patients with community-acquired pneumonia, when it is suggestive of legionellosis. Diarrhea can also occur in patients with systemic bacterial infections. In addition, although diarrhea is rare in patients with early Lyme borreliosis, the incidence is higher in those with other tick-borne infections, such as ehrlichiosis, tick-borne relapsing fever and Rocky Mountain spotted fever. Unfortunately, it is often not established whether diarrhea is an initial symptom or develops during the course of the disease. The real incidence of diarrhea in some infectious diseases must also be questioned because it could represent an adverse reaction to antibiotics. PMID- 16265205 TI - Therapy insight: Management of hepatitis C in patients coinfected with HIV. AB - Infection with HCV is common in HIV-infected patients and is an increasingly important public health problem. The medical management of hepatitis C in HIV infected patients is complicated by immune suppression, potential drug interactions and toxicities, and the relative paucity of health-care providers with expertise in the management of both diseases. Nonetheless, there are now data to support the safety, tolerability and efficacy of hepatitis C treatment with peginterferon plus ribavirin in HIV-infected patients, and the impetus to treat these patients is, therefore, strong. Although the standard of care for the treatment of hepatitis C in HIV-infected patients has been more clearly defined, the delivery of care for hepatitis C remains inconsistent in many settings. The development and implementation of single-center multidisciplinary programs that combine the expertise of HIV specialists, hepatologists, gastroenterologists, psychiatrists, and addiction specialists, are needed to improve hepatitis C treatment outcomes in HIV-infected patients. This review considers the management of HCV infection in HIV-infected patients. PMID- 16265206 TI - Therapy insight: Osteoporosis in inflammatory bowel disease--advances and retreats. AB - There is a discrepancy between the high rates of reduced bone mineral density (BMD) reported in patients with inflammatory bowel disease (IBD) and the relatively low fracture rates observed in population-based studies. When fractures occur, they are most common among the elderly IBD population. It has become clear that BMD is but one of several important factors to be considered when assessing fracture risk. Ideally, BMD should be assessed selectively, as opposed to assessing this measure in all IBD patients simply because they carry an IBD diagnosis. Preventing bone loss should begin with an attempt to limit corticosteroid-induced bone loss. This can be done by using the minimum effective prednisolone dose, substituting budesonide when appropriate, administering other steroid-sparing immunomodulators, or by prescribing additional agents that enhance bone health. The administration of calcium and vitamin D appears to maintain or enhance bone mass. Bisphosphonates are of unclear additional benefit to the majority of patients who are at low fracture risk. Although more data are required to understand the best strategy to prevent fractures, a greater appreciation of the role of selective BMD testing and the utility of simple therapeutic strategies (such as calcium and vitamin D supplements) is emerging. PMID- 16265208 TI - Obesity and visceral fat: a growing inflammatory disease. PMID- 16265207 TI - Massive lower gastrointestinal bleeding after rejection of pancreatic transplants. AB - BACKGROUND: This article highlights two cases of massive lower gastrointestinal bleeding in patients on dialysis after rejection of kidney-pancreas transplants. Patient 1 was a 34-year-old female with 27 years of type I diabetes, who had a kidney-pancreas transplant in 1996, which was complicated by rejection of the kidney and pancreas in 2000 and 2002, respectively. Later in 2002, she presented in shock after experiencing cramping abdominal pain and passage of large bloody stools. Patient 2 was a 38-year-old male with 26 years of type I diabetes, who had a pancreas-kidney transplant in 1998, which was complicated by rejection of the kidney and pancreas in early 2003. He presented in late 2003 with a single episode of coffee-ground emesis and two episodes of brisk hematochezia. INVESTIGATION: Arterial angiography. DIAGNOSIS: Pseudoaneurysm and small-bowel fistula from the arterial supply to the transplanted pancreas. MANAGEMENT: Angiographic embolization of the aneurysmal vessel and fistula achieved hemostasis. Patient 1 did not have her transplanted organ surgically removed and suffered a recurrent massive lower gastrointestinal bleed that proved fatal. In Patient 2, subsequent surgery and removal of the rejected pancreas was performed and the patient continues to do well. PMID- 16265209 TI - Is there still a role for surgery in bleeding portal hypertension? PMID- 16265224 TI - Does nadolol improve the efficacy of endoscopic variceal ligation in the treatment of variceal bleeding? PMID- 16265225 TI - Does bare-stent TIPS increase the risk of hepatocellular carcinoma in patients with cirrhosis? PMID- 16265226 TI - How accurate is the Cleveland Clinic Foundation model in predicting operative risk in colorectal cancer patients? PMID- 16265227 TI - Can dietary oligofructose prevent antibiotic-associated diarrhea? PMID- 16265228 TI - Can Saccharomyces boulardii prevent antibiotic-associated diarrhea in children? PMID- 16265229 TI - Is acarbose an effective drug for treating patients with cirrhosis and hepatic encephalopathy? PMID- 16265230 TI - Therapy insight: Vascular complications in patients with inflammatory bowel disease. AB - Inflammatory bowel disease (IBD) is associated with an increased risk of vascular complications. The most important of these complications are arterial and venous thromboembolism, which represent a significant cause of morbidity and mortality in IBD patients. Recent data suggest that thromboembolism is a disease-specific extraintestinal manifestation of IBD. The most common thrombotic manifestations in IBD are deep vein thrombosis of the leg and pulmonary emboli. It has been suggested that disease activity and the extent of colonic localization are correlated with the risk of developing thromboembolism. The occurrence of thrombosis in patients with IBD is partially attributed to the existing hypercoagulable state in IBD. Both coagulation and fibrinolysis are activated in patients with IBD; this is especially true for those with active disease. The most common risk factors for thrombophilia in IBD patients with venous thromboembolism are Leiden mutation in the gene encoding factor V, hyperhomocysteinemia, and antiphospholipid antibodies. The main genetic defects that have been established as risk factors for venous thrombosis are rather uncommon in IBD, but when present increase the risk of thromboembolism. Screening for coagulation defects seems justified only in IBD patients with a history of thrombosis or a family history of venous thromboembolic events. Antithrombotic treatment of IBD patients with venous thromboembolism is similar to that of thrombotic non-IBD patients. PMID- 16265231 TI - Mechanisms of disease: adipocytokines and visceral adipose tissue--emerging role in nonalcoholic fatty liver disease. AB - There is increasing evidence that visceral adipose tissue is a causative risk factor for fatty liver and nonalcoholic steatohepatitis. Adipose tissue-derived secretory proteins are collectively named adipocytokines. Obesity and mainly visceral fat accumulation impair adipocyte function and adipocytokine secretion and the altered release of these proteins contributes to hypertension, impaired fibrinolysis and insulin resistance. This review summarizes recent findings on the role of the adipocytokines adiponectin, leptin and resistin in the context of hepatic insulin resistance, fatty liver and liver fibrosis. Elevated levels of resistin antagonize hepatic insulin action and raise plasma glucose levels. Leptin exerts insulin-sensitizing effects, but obesity has been linked to leptin resistance and low levels of circulating leptin receptor, indicating that high levels of leptin cannot mediate its beneficial effects. Adiponectin improves insulin sensitivity; however, low circulating adiponectin is found in the obese state. Adiponectin is an anti-inflammatory protein, whereas leptin augments inflammation and fibrogenesis. Disturbed adipocytokine secretion might, therefore, promote hepatic steatosis and the development of nonalcoholic steatohepatitis. The beneficial effects of the therapeutic approaches so far tested in the treatment of fatty liver disease and fibrosis might be due to the modulation of these adipocytokines. PMID- 16265232 TI - Superior mesenteric vein thrombosis after colectomy in a patient with Crohn's disease. AB - BACKGROUND: An 18-year-old woman with a history of Crohn's disease presented in January 2004 with severe epigastric pain, nausea and vomiting of 4 hours' duration. The patient was diagnosed with inflammatory bowel disease, thought to be consistent with ulcerative colitis, in March 2003, but had no medical history up until this point. Initial treatment with mesalamine was unsuccessful and she subsequently presented with medically resistant fulminant colitis and required an urgent colectomy in June 2003. Her immediate postoperative course was uneventful and she was discharged on tapering doses of prednisone. In August 2003, an ileoscopy revealed inflamed, mildly ulcerated mucosa, and biopsies were consistent with Crohn's disease. Azathioprine was added to the treatment regimen and the patient tapered off prednisone. At this stage the patient continued to do well clinically up until presentation. INVESTIGATIONS: Small bowel series, abdominal CT scan, abdominal ultrasound, exploratory laparotomy. DIAGNOSIS: Acute mesenteric ischemia secondary to superior mesenteric vein thrombosis. MANAGEMENT: Resection of necrotic bowel, antibiotics and systemic anticoagulation. PMID- 16265233 TI - Why do we need yet another journal? PMID- 16265234 TI - New strategies to expand the donor pool: are they misguided? PMID- 16265249 TI - Can cardiac-resynchronization therapy reduce mortality in patients suffering from advanced chronic heart failure? PMID- 16265250 TI - Is pulmonary vein antrum isolation safe and effective in valvular heart disease and open heart surgery? PMID- 16265251 TI - Can beta-blocker use lower mortality and improve myocardial tissue recovery after acute myocardial infarction? PMID- 16265252 TI - Do implantable cardioverter defibrillators lower mortality risk in patients with cardiomyopathy? PMID- 16265253 TI - Can pravastatin lower coronary event rate if HDL and LDL cholesterol are at low levels? PMID- 16265254 TI - Does carotid endarterectomy prevent disabling and fatal strokes in neurologically asymptomatic subjects? PMID- 16265255 TI - Can delayed enhancement and T2-weighted imaging distinguish acute from chronic myocardial infarction? PMID- 16265256 TI - The role of sodium in hypertension is more complex than simply elevating arterial pressure. AB - Excessive salt intake exacerbates hypertension and further increases left ventricular mass in clinical essential and experimental hypertension. Additionally, a growing body of evidence strongly suggests that high dietary salt loading exerts detrimental cardiac effects independently of its hemodynamic load. The clinical evidence of cardiac structural and functional alterations associated with salt is, however, scarce. In order to explore the purported beliefs in humans, in this review we draw on our experimental studies in naturally occurring hypertension and discuss the clinical implications of the nonhemodynamic mechanisms underlying these salt-related changes. PMID- 16265257 TI - Therapy insight: aortic aneurysm and dissection in Marfan's syndrome. AB - Aortic dissection and aneurysm are common clinical problems with life-threatening consequences; they are also the hallmark of several genetic diseases, including Marfan's syndrome (MFS). In spite of clinical and surgical advances that have increased life expectancy for affected patients, cardiovascular manifestations remain significant contributors to morbidity and mortality in MFS. Dissecting aortic aneurysm in this disorder is accounted for by mutations in fibrillin-1, the major component of the microfibrils associated with elastin in the elastic fibers of the aortic media. Genetic studies of human patients and murine models have yielded invaluable insights into the pathophysiology of aneurysm formation and progression in MFS. They have also revealed a previously unappreciated role of microfibrils in regulating transforming growth factor and bone morphogenetic protein signaling. As a result, exciting new hypotheses have emerged regarding the pathogenesis of MFS, as well as opportunities to explore translational applications of this information that may be relevant to various manifestations of the disease. PMID- 16265258 TI - Mechanisms of disease: new mechanisms of antiarrhythmic actions. AB - Cardiac arrhythmias are a leading cause of morbidity and mortality in many developed countries. Despite intensive investigation, the cellular mechanisms for most cardiac arrhythmias have not been clearly established. As a consequence, drug therapy for most forms of atrial and ventricular arrhythmias remains largely empirical and ineffective, leading to the increased use of nonpharmacologic treatments. Clearly, new approaches to the prevention of cardiac arrhythmias are needed. Here we review the current experimental basis for several promising antiarrhythmic strategies, with a focus on those targeted against atrial and ventricular fibrillation. Although none of these strategies is yet ready for clinical application, they provide a basis for cautious optimism that effective pharmacologic therapy for fatal cardiac rhythm disturbances could be forthcoming. PMID- 16265259 TI - Mechanisms of disease: local renin-angiotensin-aldosterone systems and the pathogenesis and treatment of cardiovascular disease. AB - Accumulating evidence has made it clear that not only does the renin-angiotensin aldosterone system (RAAS) exist in the circulation where it is driven by renal renin, but it is also active in many tissues-and likely within cells as well. These systems might not be completely independent of each other, but rather interact. These local RAASs affect tissue and cellular angiotensin II concentrations and appear to be associated with clinically relevant physiologic and pathophysiologic actions in the cardiovascular system and elsewhere. Evidence in support of this possibility is reviewed here. In addition, direct (pro)renin action after binding to its specific receptor, the existence of renin transcripts, which apparently encode an intracellular renin, the discovery of an angiotensin-converting-enzyme homologue (ACE2), which leads to enhanced generation of angiotensin-(1-7) and the newly appreciated role of angiotensin receptor dimerization in the regulation of angiotensin activity, all point to the conclusion that the RAASs are complexly regulated, multifunctional systems with important roles both within and outside the cardiovascular system. PMID- 16265261 TI - Atrial fibrillation, sleep apnea and obesity. AB - BACKGROUND: A 60-year-old male with obesity (body-mass index 43 kg/m(2)) presented with recurrent symptomatic atrial fibrillation (AF), which he had had since age 41 years. The AF was refractory to treatment with antiarrhythmic drugs. Pacemaker implantation for tachycardia-bradycardia syndrome was required as well as ablation for atrial flutter, and the patient underwent a total of four DC cardioversions. Sleep studies showed mild to moderate obstructive sleep apnea, but continuous positive airway pressure was not tolerated. Pacemaker interrogations demonstrated mode-switch episodes, indicating continuing AF. He was scheduled for catheter ablation targeting pulmonary vein antral isolation. He embarked on a weight-loss program, which successfully reduced AF burden. INVESTIGATIONS: Echocardiography, stress testing, polysomnography, pacemaker interrogations and C-reactive protein. DIAGNOSIS: AF, atrial flutter, tachycardia bradycardia syndrome, obstructive sleep apnea and morbid obesity. MANAGEMENT: Antiarrhythmic drug therapy, DC cardioversion, anticoagulation, atrial flutter ablation, permanent pacemaker implantation, continuous positive airway pressure and weight loss. PMID- 16265262 TI - Institutional review boards: the cost to academic medical centers. PMID- 16265260 TI - Technology insight: targeting of biological molecules for evaluation of high-risk atherosclerotic plaques with magnetic resonance imaging. AB - Identification of high-risk atherosclerotic lesions prone to rupture and thrombosis may greatly decrease the morbidity and mortality associated with atherosclerosis. The development of magnetic resonance imaging contrast agents that specifically target components of the atherosclerotic plaque might enable non-invasive detection of high-risk lesions. This review discusses a variety of molecules present in atherosclerotic plaque that could serve as targets for specific contrast agents. Ultimately, such agents may allow the identification of high-risk atherosclerotic lesions in patients and enable treatment of these patients before lesion progression and complications. PMID- 16265263 TI - Is population screening for colorectal cancer cost-effective? PMID- 16265264 TI - Is a life-long gluten-free diet for patients with celiac disease successful? PMID- 16265279 TI - Is endoscopic balloon therapy an effective treatment for patients with Crohn's disease strictures? PMID- 16265280 TI - Does procedural volume matter in the efficacy of self-expanding stents in patients with esophageal cancer? PMID- 16265281 TI - Is radiofrequency ablation an effective long-term treatment for early-stage hepatocellular carcinoma? PMID- 16265282 TI - Is Bifidobacterium a more effective probiotic therapy than Lactobacillus for patients with irritable bowel syndrome? PMID- 16265283 TI - How successful is intestinal transplantation and what improves graft survival? PMID- 16265284 TI - Mechanisms of disease: vitamin D and inflammatory bowel disease. AB - Until recently, 1,25-dihydroxyvitamin D(3) (1,25(OH)(2)D(3))-the active form of vitamin D-was thought to function primarily as a regulator of calcium and phosphate metabolism. More diverse functionality was indicated by the discovery of the vitamin D receptor in tissues that are not involved in calcium and phosphate homeostasis. Detection of the vitamin D receptor in monocytes and activated T cells has sparked interest in the immunomodulatory properties of vitamin D. Here, we review the role of vitamin D in regulation of the immune system, and evidence for its involvement in the pathogenesis of inflammatory bowel disease. PMID- 16265285 TI - Diagnosis and management of intraductal papillary mucinous neoplasia. AB - Intraductal papillary mucinous neoplasia (IPMN) encompasses a spectrum of neoplastic change in the epithelial lining of the pancreatic duct. As such, it is difficult to predict the presence of malignancy, and pancreatic resection is established as the only effective treatment for IPMN. There is controversy about the natural history, evaluation, surgical management and surveillance of IPMN; as IPMN is being increasingly recognized, this controversy is being brought to the forefront of clinical practice. This review presents pooled surgical data, future directions and a proposed algorithm for the management of patients with IPMN. PMID- 16265286 TI - Technology insight: ablative techniques for Barrett's esophagus--current and emerging trends. AB - New mucosal ablative techniques that can be used in the esophagus have emerged over the past two decades. These techniques have been developed primarily to treat the precursors of esophageal adenocarcinoma: dysplasia in Barrett's esophagus and early esophageal cancer. Although high-grade dysplasia and early stage cancer can be treated with esophagectomy, the inherent morbidity and mortality of esophageal adenocarcinoma and the morbidities, difficulties, costs and limitations of the current technology mean that there has been a significant increase in interest and research regarding alternative treatments such as ablative techniques. At this stage it is not clear which of the numerous endoscopic ablative techniques available-photodynamic therapy, laser therapy, multipolar electrocoagulation, argon plasma coagulation, endoscopic mucosal resection, radiofrequency ablation or cryotherapy-will emerge as superior. In addition, it has yet to be determined whether the risks associated with ablation therapy are less than the risk of Barrett's esophagus progressing to cancer. Whether ablation therapy eliminates or significantly reduces the risk of cancer, eliminates the need for surveillance endoscopy, or is cost-effective, also remains to be seen. Comparative trials that are now underway should help to answer these questions. PMID- 16265287 TI - Diagnosis and treatment of a patient with gallstone ileus. AB - BACKGROUND: A 79-year-old white woman presented with upper abdominal pain. She had a history of rheumatoid arthritis since she was 19 years old, which was treated with prednisolone, leflunomide, diclofenac and pantoprazole. She also had factor VII deficiency. The patient had been hospitalized 2 months previously with sepsis presumed to be due to urinary infection, and was treated with antibiotics. Sonography at this time revealed a gallbladder with a monstrous thick wall and stones, and the first differential diagnosis was cholecystitis. Cholecystectomy was planned after amelioration of the patient's general state, but her general state worsened. INVESTIGATIONS: Sonography, endoscopy of the upper and lower intestine, and CT scan. DIAGNOSIS: Biliodigestive fistula and gallstone ileus. MANAGEMENT: Enterolithotomy, stenting, endoscopic retrograde cholangiopancreatography, and surgery. PMID- 16265288 TI - Cell therapy for acute myocardial infarction: where are we heading? PMID- 16265289 TI - Percutaneous valve replacement: a fad or an advance? PMID- 16265304 TI - Cardiac imaging in the emergency room: a practical method to identify acute myocardial infarction? PMID- 16265305 TI - What factors predict pacemaker syndrome and how can it be treated in patients with sinus node dysfunction? PMID- 16265307 TI - Have recent statin trial findings affected the National Cholesterol Education Program guidelines? PMID- 16265306 TI - Is mortality raised in patients receiving percutaneous coronary intervention if they suffer from anemia? PMID- 16265308 TI - Can familial combined hyperlipidemia diagnostic criteria be improved by the use of a nomogram? PMID- 16265309 TI - Is antithrombotic therapy a risk-free and beneficial treatment for patients with heart failure? PMID- 16265310 TI - Does the time between ventricular assist device implantation and heart transplantation affect survival? PMID- 16265312 TI - Myocardial-cell replacement: the science, the clinic and the future. AB - The traditional view of the heart is of an organ incapable of self-renewal. Hypotheses on the genesis of pump dysfunction in heart failure include emerging concepts of myocyte deficiency due to attritional ischemia and chronic apoptotic cell loss. In the adult heart, inadequate regenerative capacity was presumed to exist to counterbalance such extensive myocyte depletion. Preliminary animal and human studies now challenge the paradigm of myocardial regenerative inadequacy, with findings suggesting that noncardiac cells implanted in the dysfunctional heart augment myocyte deficiency and contractile mass. Data from these studies remain inconclusive and have generated much debate in the basic science and clinical communities. Controversial issues center on the scientific basis for regeneration in the heart, the mechanism of cell-therapy benefit and the safety and appropriateness of clinical trials based on these concepts. This review will evaluate the scientific basis for myocardial-cell replacement, with emphasis on current experimental and human data. We will explore unresolved questions of experimental design, mechanism of action, therapeutic strategies and safety concerns in an era of rising numbers of human cell-therapy trials. Prospects for more widespread clinical application of myocardial-cell replacement and future hurdles to be overcome in this field will also be addressed. PMID- 16265311 TI - Therapeutic approaches to raising plasma HDL-cholesterol levels. AB - Epidemiologic data from the Framingham and Prospective Cardiovascular Munster studies, demonstrating an inverse correlation between the plasma concentration of HDLs and the incidence of cardiovascular disease, have driven research to explore precisely how HDLs confer this cardioprotective effect. HDLs are anti inflammatory, antithrombogenic and have vasoactive effects, as well as being efficient cholesterol acceptors enabling the removal of cholesterol from peripheral tissues, all functions that are likely to protect the vasculature. The first part of this article will review the clinical evidence in support of the pleiotropic effects of HDLs, along with laboratory-based investigations of the molecular mechanisms of action. As the evidence of clinical benefits of raising plasma HDL concentration has increased, so has the number of strategies currently being considered to achieve this goal. The second part of this article will review three current strategies: infusion of HDL-like products, comparing physicopharmacologic characteristics of the two commercial products currently under trial; the use of fibrates to raise plasma HDLs (although fibrates primarily reduce triglyceride levels, certain derivatives are able to induce significant increases in plasma HDLs); and the use of drugs that inhibit cholesterol ester transfer protein (these drugs increase plasma HDL concentration either alone or as an adjunct therapy with statins). The clinical efficacy and mechanism of action of fibrates and inhibitors of cholesterol ester transfer protein will be reviewed. PMID- 16265313 TI - Ischemic preconditioning and preinfarction angina in the clinical arena. AB - In animals, brief episodes of ischemia before a total coronary occlusion protect the heart and result in a smaller myocardial infarct size. In humans, episodes of angina before acute myocardial infarction might also confer a preconditioning or protective effect; numerous studies show that preinfarction angina is associated with smaller infarcts. Preinfarction angina is also associated with reductions in ventricular dysfunction, arrhythmias and incidence of no-reflow phenomena, and, in some cases, improved survival. The protective effect of preconditioning in humans is characterized by marked individual variations and seems to be attenuated in women, people with diabetes and the elderly. Exercise seems to be an important way to induce preconditioning in humans and preserves it in the elderly. PMID- 16265314 TI - Drug insight: If inhibitors as specific heart-rate-reducing agents. AB - Heart rate is determined primarily by spontaneously repeating net inward current carried by sodium ions and potassium ions through hyperpolarization-activated cyclic-nucleotide-gated channels. Within the heart, these channels are found most abundantly in sinoatrial cardiomyocytes. The channels open in response to membrane hyperpolarization, modulated by local cAMP concentrations. They permit activation of the I(f) current, which can be blocked specifically by molecules characterized by linked benzazepinone and benzocyclobutane rings, and which are devoid of effects on cardiac conduction, inotropy or peripheral vascular tone. The resulting heart-rate reduction has been effective in angina prevention in clinical trials involving 4,000 patients, using the prototype I(f) inhibitor, ivabradine. No serious adverse events have been attributed to the treatment; the most prominent side-effect is dose-related, always reversible and often transient visual symptoms that seldom result in voluntary drug discontinuation. PMID- 16265331 TI - Obesity: a challenge for science and society. PMID- 16265315 TI - Mechanisms of disease: roles of nitric oxide and endothelin-1 in delayed cerebral vasospasm produced by aneurysmal subarachnoid hemorrhage. AB - Molecular biologic investigations in the past decades have begun to unravel the intracellular mechanisms involved in vasomotor regulation of cerebral blood vessels and their failure in delayed cerebral vasospasm produced by aneurysmal subarachnoid hemorrhage. Progress in deciphering macrovascular regulatory mechanisms and their failure in delayed cerebral vasospasm induced by aneurysmal subarachnoid hemorrhage have revealed that there are at least two important vasoactive substances-nitric oxide and endothelin-1-that play important roles in the clinical manifestations of subarachnoid-hemorrhage-induced cerebral vasospasm. Nitric oxide is a cell-membrane-permeable free radical gas that accounts for the phenomenon of vasodilatation by a variety of vasodilator agents. Endothelin-1, a 21 amino acid peptide, is one of the most potent constricting factors. Cerebral vasospasm is thought to represent a disturbance in the cerebral vasomotor equilibrium for which these two physiologically antagonistic compounds are at least partly responsible. Advances in our understanding of the molecular responses of the cerebral vasculature to subarachnoid hemorrhage should lead to more comprehensive management as knowledge becomes translated into development of effective pharmacologic agents to reverse or prevent cerebral vasospasm following aneurysmal subarachnoid hemorrhage. PMID- 16265333 TI - Does intensive statin therapy lower mortality and cardiovascular event risk in patients with acute coronary syndrome? PMID- 16265332 TI - Pharmacogenetics: the outlook for genetic testing in statin therapy. PMID- 16265334 TI - Fetal echocardiography: can early imaging identify fetuses at risk of congenital heart disease? PMID- 16265335 TI - Should elderly heart failure patients be treated with beta-blockade therapy? PMID- 16265336 TI - Should pacemakers be removed in the event of localized implantation-site infection? PMID- 16265337 TI - Can aortic atherosclerotic plaques predict cerebrovascular and cardiac events? PMID- 16265338 TI - Is the risk of sudden death and nonfatal cardiac arrest reduced by ACE inhibitor therapy? PMID- 16265339 TI - Early reperfusion strategies after acute ST-segment elevation myocardial infarction: the importance of timing. AB - Acute ST-segment elevation myocardial infarction is estimated to occur in more than 500,000 people in the US every year. With the introduction of reperfusion therapy by fibrinolysis or primary angioplasty, mortality has significantly fallen. Although fibrinolysis is more readily available than primary angioplasty, the latter is more effective and results in better short-term and long-term outcomes if performed in a timely manner by an experienced operator and hospital team. The ischemic time, door-to-balloon time and clinical risk are important determinants of favorable outcome. Primary angioplasty is the preferred reperfusion strategy when symptom onset is longer than 3 h, in high-risk patients, such as those with cardiogenic shock, congestive heart failure or elderly age, and those with contraindications for fibrinolysis. Primary angioplasty is the preferred strategy in interventional facilities, with a goal door-to-balloon time of less than 90 min. For patients who present to noninterventional facilities, transfer to a hospital capable of primary angioplasty is safe and effective if the additional treatment delay is less than 90 min. Facilitated percutaneous coronary intervention has been shown in several small trials to offer early vessel patency and improve outcomes compared with fibrinolysis alone, but has not been shown to reduce mortality. Larger trials are ongoing to evaluate the benefit of this approach. The establishment of an effective and efficient system for the rapid transport of patients to centers capable and experienced in primary angioplasty is severely needed to provide optimum treatment and outcomes to patients with ST-segment elevation acute myocardial infarction. PMID- 16265340 TI - C-reactive protein comes of age. AB - Cardiovascular disease remains a leading cause of death throughout the world despite advances in its detection and treatment. Commonly used risk algorithms, such as the Framingham Risk Score fail to identify all affected individuals. Novel cardiovascular risk factors that identify these missed individuals would greatly improve overall care of patients. C-reactive protein (CRP), an inflammatory biomarker, has emerged as a leading candidate to fulfill this role. Based on the results of several prospective epidemiologic studies, CRP has emerged as one of the most powerful predictors of cardiovascular disease. This marker provides valuable information to clinicians in various clinical settings, ranging from overt cardiovascular disease, stable angina, presenting acute coronary syndromes and peripheral vascular disease, to the metabolic syndrome. Furthermore, CRP has been demonstrated to actively contribute to all stages of atherogenesis, participating in endothelial dysfunction, atherosclerotic-plaque formation, plaque maturation, plaque destabilization and eventual rupture. Thus, it might also serve as a therapeutic target. It is our contention that the future will see much wider use of CRP and CRP-driven therapies in clinical medicine, improving our ability to identify and manage cardiovascular disease. PMID- 16265341 TI - Cell therapy for cardiovascular disease: what cells, what diseases and for whom? AB - Experimental and human data suggesting progenitor cells possess the capacity to regenerate tissue and augment repair in injured organs has generated widespread interest in the basic research and clinical communities. Nowhere have these findings been more tantalizing than in human cardiovascular disease, in which vasculogenesis and myocardial regeneration logically and understandably remain as attractive therapeutic targets. Burgeoning experimental evidence attests to the proangiogenic, vasculogenic and tissue reparative capabilities of a broad range of progenitor cells derived from the bone marrow, circulation and a number of other tissues in vivo. Studies demonstrating the most apparent therapeutic success are those implicated in revascularization and repair of acute or chronically ischemic tissues in the heart and the peripheral vascular system. Numerous small clinical trials have yielded promising preliminary results without clear evidence of a superiority for a specific cell type or clinical disease entity as the most suitable target for cell therapy. This review will evaluate the scientific rationale for use of a specific cell or cells, the cardiovascular disease states most appropriate for targeted cell therapy, and the patient specific barriers to therapeutic success, including emerging hurdles such as cardiovascular risk factors and comorbidities in eligible subjects. PMID- 16265342 TI - The sinus tachycardias. AB - Sinus tachycardia, in the forms of four distinct rhythm disturbances, is frequently encountered in clinical practice but is often overlooked. The most common rhythm, normal sinus tachycardia, whether physiologic, pathologic or iatrogenic, is predominantly catecholamine driven, is virtually asymptomatic and is managed by identifying and treating the underlying cause. The other so-called primary sinus tachycardias, which include inappropriate sinus tachycardia, postural orthostatic tachycardia syndrome and sinus node re-entry tachycardia, have fundamentally different clinical features, basic underlying etiologic mechanisms and treatment strategies. Differentiation of these types from normal sinus tachycardia and from other atrial arrhythmias is crucial for successful management. Accurate diagnosis and appropriate therapy of the sinus tachycardias not only prevents multiple consultations but might also have important long-term prognostic implications. PMID- 16265343 TI - Reversible cardiomyopathy caused by administration of interferon alpha. AB - BACKGROUND: A 56-year-old man with normal cardiac function received treatment with interferon alpha-2b for malignant melanoma. Eight months after the initiation of therapy he developed fatigue and dyspnea on exertion. Two months later, he was admitted to hospital with ORTHOPNEA, worsening dyspnea and cough. Physical examination findings were consistent with congestive heart failure. Laboratory studies were notable for hypothyroidism. Echocardiography revealed severe, global left-ventricular dysfunction. INVESTIGATIONS: Echocardiogram, electrocardiogram, serum chemistries, coronary angiography, right-heart and left heart catheterization and endomyocardial biopsy. DIAGNOSIS: Interferon alpha-2b induced cardiomyopathy. MANAGEMENT: Intravenous dobutamine and dopamine for cardiogenic shock and discontinuation of interferon alpha-2b. PMID- 16265344 TI - Phase II randomised trial of raltitrexed-oxaliplatin vs raltitrexed-irinotecan as first-line treatment in advanced colorectal cancer. AB - The purpose of this phase II randomised trial was to determine which of two schemes, raltitrexed-irinotecan or raltitrexed-oxaliplatin, offered better activity and less toxicity in patients with advanced colorectal cancer (CRC). A total of 94 patients with previously untreated metastatic CRC were included and randomised to receive raltitrexed 3 mg m(-2) followed by oxaliplatin 130 mg m(-2) on day 1 (arm A), or CPT-11 350 mg m(-2) followed by raltitrexed 3 mg m(-2) (arm B). In both arms treatment was repeated every 3 weeks. Intent-to-treat (ITT) analysis showed an overall response rate of 46% (95% CI, 29.5-57.7%) for arm A, and 34% (95% CI, 19.8-48.4%) for arm B. Median time to progression was 8.2 months for arm A and 8.8 months for arm B. After a median follow-up of 14 months, 69% of patients included in arm A were still alive, compared to 59% of those included in arm B. Overall, 31 patients (65%) experienced some episode of toxicity in arm A and 32 patients (70%) in arm B, usually grade 1-2. The most common toxicity was hepatic, with 29 patients (60%) in arm A and 24 patients (62%) in arm B, and was grade 3-4 in four (8%) and four (9%) patients, respectively. In all, 14 patients (29%) from arm A and 24 patients (52%) from arm B had some grade of diarrhoea (P<0.03). Neurologic toxicity was observed in 31 patients (64%) in arm A, and was grade 3-4 in five patients (10%), while a cholinergic syndrome was detected in nine patients (19%) in arm B. There were no differences in haematologic toxicity. One toxic death (2%) occurred in arm A and three (6.5%) in arm B. In conclusion, both schemes have high efficacy as first-line treatment in metastatic CRC and their total toxicity levels are similar. Regimens with raltitrexed seem a reasonable alternative to fluoropyrimidines. PMID- 16265345 TI - Circadian function in patients with advanced non-small-cell lung cancer. AB - This study aimed to evaluate whether patients with advanced non-small-cell lung cancer experience disrupted rest-activity daily rhythms, poor sleep quality, weakness, and maintain attributes that are linked to circadian function such as fatigue. This report describes the rest-activity patterns of 33 non-small-cell lung cancer patients who participated in a randomised clinical trial evaluating the benefits of melatonin. Data are reported on circadian function, health related quality of life (QoL), subjective sleep quality, and anxiety/depression levels prior to randomisation and treatment. Actigraphy data, an objective measure of circadian function, demonstrated that patients' rest-activity circadian function differs significantly from control subjects. Our patients reported poor sleep quality and high levels of fatigue. Ferrans and Powers QoL Index instrument found a high level of dissatisfaction with health-related QoL. Data from the European Organization for Research and Treatment for Cancer reported poor capacity to fulfil the activities of daily living. Patients studied in the hospital during or near chemotherapy had significantly more abnormal circadian function than those studied in the ambulatory setting. Our data indicate that measurement of circadian sleep/activity dynamics should be accomplished in the outpatient/home setting for a minimum of 4-7 circadian cycles to assure that they are most representative of the patients' true condition. We conclude that the daily sleep/activity patterns of patients with advanced lung cancer are disturbed. These are accompanied by marked disruption of QoL and function. These data argue for investigating how much of this poor functioning and QoL are actually caused by this circadian disruption, and, whether behavioural, light-based, and or pharmacologic strategies to correct the circadian/sleep activity patterns can improve function and QoL. PMID- 16265346 TI - Neuronal characteristics of small-cell lung cancer. AB - Wide ranging experimental evidence suggests that human small-cell lung cancer (SCLC) has a number of molecular and subcellular characteristics normally associated with neurones. This review outlines and discusses these characteristics in the light of recent developments in the field. Emphasis is placed upon neuronal cell adhesion molecules, neurone-restrictive silencer factor, neurotransmitters/peptides and voltage-gated ion, especially Na(+) channels. The hypothesis is put forward that acquisition of such characteristics and the membrane 'excitability' that would follow can accelerate metastatic progression. The clinical potential of the neuronal characteristics of SCLC, in particular ion channel expression/activity, is discussed in relation to possible novel diagnostic and therapeutic modalities. PMID- 16265348 TI - Human papillomavirus 16 infection in adenocarcinoma of the cervix. AB - The impact of the success of organised cervical screening programme results in a steady decline of the incidence of squamous cell carcinoma of the cervix but a concomitant increase in the incidence of the less common histological subtypes, particularly adenocarcinoma of the cervix (ACC). Although Human papillomavirus (HPV) infection is believed to be a necessary cause of cervical cancer, its role in the pathogenesis of ACC is not well established. Established associations between oncogenic strains of HPV and ACC are based on molecular studies carried out on entire tumour block sections. In this study, the cervical adenocarcinoma cells of a 10-year cohort of women diagnosed with ACC were dissected using the PixCell II Laser Microdissecting System to detect the HPV 16 genome sequence using the real-time quantitative polymerase chain reaction to confirm the presence of HPV DNA within ACC cells. By coupling these two sophisticated techniques, the HPV DNA copy number cell could be calculated to investigate its role. The prevalence of HPV 16 infection in this cohort was 24%, which is significantly higher than the control group (chi(2), P=0.014). Women with ACC also had significantly higher HPV DNA copy number per cell compared to the control group (P=0.00007). Higher HPV DNA copy number is associated with risk of developing ACC. PMID- 16265347 TI - S100A4 (Mts1) gene overexpression is associated with invasion and metastasis of papillary thyroid carcinoma. AB - Tumour cell invasion and metastasis are the hallmark of malignant neoplasm. S100A4 is a member of small calcium-binding protein family and is involved in the cell proliferation and cancer progression. S100A4 is capable of inducing metastasis in animal models and is associated with aggressive phenotype of human tumours. We previously identified S100A4 as a candidate gene involved in anaplastic thyroid cancer metastasis by microarray analysis. To further determine whether S100A4 overexpression is associated with thyroid tumour invasion and metastasis, in the present study, we examined S100A4 gene expression in six benign multinodular goitres (MNG) and 28 matched samples of adjacent normal thyroid tissue (N), primary (T) and metastatic (M) papillary thyroid carcinomas (PTC) by immunohistochemistry and real-time reverse transcription-polymerase chain reaction (RT-PCR) analysis. This gave us the advantage of directly comparing levels of S100A4 expression within the same genetic background. Using immunohistochemistry, we found that high levels of S100A4 were detected in 24 of 28 (86%) PTC specimens and their local regional lymph node or distant metastases. No S100A4 staining was observed in normal thyroid tissues and simple MNG. However, in MNG coexistent with PTC, moderate focal staining could be found in 11 of 15 MNG adjacent to PTC. The S100A4 was stained more intensely in invading fronts than in central portions of both T and M. Real-time RT-PCR analysis of primary tumours and their matched lymph node metastasis demonstrated that significantly higher S100A4 transcripts were present in metastatic tumours as compared to the primary tumours (P<0.01). These data suggest that overexpression of S100A4 is associated with thyroid tumour invasion and metastasis and it may be a potential target for therapeutic intervention. PMID- 16265349 TI - RASSF2, a potential tumour suppressor, is silenced by CpG island hypermethylation in gastric cancer. AB - RASSF2, a member of the RASSF1 family, has recently been identified as a potential tumour suppressor. We examined methylation status in multiple regions which included the CpG island and spanned the transcription start site of RASSF2 in 10 gastric cancer cell lines, as well as 78 primary gastric cancers and corresponding non-neoplastic gastric epithelia. Hypermethylation of RASSF2 in at least one of the regions examined was detected in seven (70%) of the 10 cell lines; two (20%) exhibited hypermethylation in all the regions examined including the transcription start site and lost expression of RASSF2 mRNA, which could, however, be restored by 5-aza-2' deoxycytidine treatment, while the other five (50%) cell lines exhibited hypermethylation at the 5'- and/or 3'- edge, with four of them expressing RASSF2 mRNA. In primary gastric cancers and corresponding non neoplastic gastric epithelia, frequencies of RASSF2 methylation ranged from 29% (23 out of 78) to 79% (62 out of 78) and 3% (two out of 78) to 60% (47 out of 78), respectively, at different CpG sites examined. Methylation was frequently observed at the 5'- and 3'- edges, and became less frequent near the transcription start site in both the primary gastric cancers and corresponding non-neoplastic gastric epithelia. Hypermethylation near the transcription start site was mostly cancer-specific. We thus showed that RASSF2 is silenced by hypermethylation near the transcription start site in gastric cancer. Hypermethylation was found initially to occur at the 5'- and 3'- furthest regions of the CpG island in non-neoplastic gastric epithelia, to gradually spreads near the transcription start site to shut down RASSF2 expression, and ultimately to constitute a field-defect placing tissue increased risk for development of gastric cancer. PMID- 16265350 TI - Anti-HER-2 DNA vaccine protects Syrian hamsters against squamous cell carcinomas. AB - This paper illustrates the efficacy of DNA vaccination through electroporation in the prevention of oral transplantable carcinoma in Syrian hamsters. At 21 and 7 days before tumour challenge, 19 hamsters were vaccinated with plasmids coding for the extracellular and transmembrane domains of rat HER-2 receptor (EC-TM plasmids), whereas 19 control hamsters were injected intramuscularly with the empty plasmid. Immediately following plasmid injection, hamsters of both groups received two square-wave 25 ms, 375 V cm(-1) electric pulses via two electrodes placed on the skin of the injection area. At day 0, all hamsters were challenged in the submucosa of the right cheek pouch with HER-2-positive HCPC I cells established in vitro from an 7,12-dimethylbenz[a]anthracene-induced oral carcinoma. This challenge gave rise to HER-2-positive buccal neoplastic lesions in 14 controls (73.37%), compared with only seven (36.8%, P<0.0027) vaccinated hamsters. In addition, the vaccinated hamsters displayed both a stronger proliferative and cytotoxic response than the controls and a significant anti-HER 2 antibody response. Most of the hamsters that rejected the challenge displayed the highest antibody titres. These findings suggest that DNA vaccination may have a future in the prevention of HER-2-positive human oral cancer. PMID- 16265351 TI - Characterisation and internalisation of recombinant humanised HMFG-1 antibodies against MUC1. AB - The humanised HMFG-1 immunoglobulin has been extensively developed as a clinical immunotherapeutic agent for MUC1 expressing tumours. We have constructed a single chain Fv (scFv) and Fab fragment from this antibody and shown that both these species retain their specificity for MUC1. The scFv was less stable and less soluble than the Fab. Detailed analyses of the binding kinetics of the whole IgG and Fab fragment show that the affinity for MUC1 synthetic peptides is low (approximately 100 nM for the IgG and 10 muM for the Fab), with particularly low but similar dissociation rate constants (0.031-0.095 s(-1)). Binding to native antigen on the cell surface is over two orders of magnitude better. Confocal immunofluorescence microscopy shows that both the IgG and Fab are internalised rapidly (the IgG is internalised within 15 min) and colocalise to early endosomes. This work provides an appreciation of the binding, internalising and trafficking kinetics, important for the development of future therapeutics based on this antibody. PMID- 16265352 TI - A meta-analysis of two randomised trials of early chemotherapy in asymptomatic metastatic colorectal cancer. AB - This report constitutes a prospectively planned meta-analysis combining two almost identical trials undertaken in Australasia and Canada to study the effect of starting chemotherapy immediately in asymptomatic patients with metastatic colorectal cancer. Patients (n=168) were randomised to receive either immediate or delayed treatment (at onset of predefined symptoms). Australasian patients received either weekly 5-fluorouracil and leucovorin (500 and 20 mg m(-2), respectively) (n=59) or the daily x 5 Mayo Clinic schedule (425 and 20 mg m(-2), respectively) (n=42). Canadian patients were treated with the Mayo schedule (n=67). Otherwise, the two studies were almost identical in design and each used the European Organisation for the Research and Treatment of Cancer (EORTC) QLQ C30 instrument for measuring quality of life (QoL). Treatment was continued until 6 months had elapsed or disease progression occurred. Low accrual led to trial suspension before the predetermined sample size for either study was reached. Median survival was not significantly better with immediate treatment (median 13.0 vs 11.0 months; hazard ratio, 1.15; 95% confidence interval (CI) 0.79-1.72; P=0.49). There was no statistically significant difference in progression-free survival (time from randomisation until first evidence of progression after chemotherapy, 10.2 vs 10.8 months; hazard ratio, 1.08; 95% CI 0.71-1.64; P=0.73). There was no difference in overall QoL or its individual domains between the two treatment strategies at baseline or at any subsequent time point. Early treatment of asymptomatic patients with metastatic colorectal cancer did not provide a survival benefit or improved QoL compared to withholding treatment until symptoms occurred. PMID- 16265354 TI - Should we afford implantable cardioverter defibrillator therapy? PMID- 16265353 TI - Gene expression profiling of noninvasive primary urothelial tumours using microarrays. AB - At present, the mechanism leading to bladder cancer is still poorly understood, and our knowledge about early events in tumorigenesis is limited. This study describes the changes in gene expression occurring during the neoplastic transition from normal bladder urothelium to primary Ta tumours. Using DNA microarrays, we identified novel differentially expressed genes in Ta tumours compared to normal bladder, and genes that were altered in high-grade tumours. Among the mostly changed genes between normal bladder and Ta tumours, we found genes related to the cytoskeleton (keratin 7 and syndecan 1), and transcription (high mobility group AT-hook 1). Altered genes in high-grade tumours were related to cell cycle (cyclin-dependent kinase 4) and transcription (jun d proto oncogene). Furthermore, we showed the presence of high keratin 7 transcript expression in bladder cancer, and Western blotting analysis revealed three major molecular isoforms of keratin 7 in the tissues. These could be detected in urine sediments from bladder tumour patients. PMID- 16265355 TI - Will heart valve tissue engineering change the world? PMID- 16265370 TI - Is antitachycardia pacing a safe and efficacious alternative to shocks for fast ventricular tachyarrhythmia treatment? PMID- 16265372 TI - Do serum B-type natriuretic peptide levels predict short-term mortality and angiographic success? PMID- 16265371 TI - Should chronic heart failure patients with reduced left-ventricular ejection fraction receive angiotensin-receptor blockers? PMID- 16265373 TI - Does surgical ventricular restoration improve survival in patients with anterior infarction congestive heart failure? PMID- 16265375 TI - Is there a link between carotid artery stiffness and risk of cerebrovascular events? PMID- 16265374 TI - Can oral sirolimus therapy prevent in-stent restenosis? PMID- 16265376 TI - Can contrast-induced nephropathy after percutaneous coronary intervention be accurately predicted with a risk score? PMID- 16265377 TI - Is computer-assisted, long-term warfarin therapy safe and efficacious for patients with nonrheumatic atrial fibrillation? PMID- 16265378 TI - Short QT syndrome: mechanisms, diagnosis and treatment. AB - Short QT syndrome is an inheritable primary electrical disease of the heart that was discovered in 1999. The disorder is characterized by an abnormally short QT interval (<300 ms) and a propensity to atrial fibrillation, sudden cardiac death or both. As in the case of long QT syndrome, more than one relevant genetic mutation has been identified that can lead to a short QT interval on electrocardiography; so far two have been identified. Shortening of the effective refractory period combined with increased dispersion of repolarization is the likely substrate for re-entry and life-threatening tachyarrhythmias. Thus far, 22 people have been classified as having short QT syndrome: 15 from the actual measurement of a short QT interval on electrocardiograms and 7 by history after they died from sudden cardiac death. Several cases, especially among children, have probably been overlooked, since the shortness of the QT interval becomes apparent only at heart rates less than 80 beats/min. The best form of treatment is still unknown, but prevention of atrial fibrillation has been accomplished by propafenone. Implantation of an implantable cardioverter defibrillator is recommended for prevention of sudden cardiac death. PMID- 16265379 TI - Mechanisms, management and future directions for reperfusion injury after acute myocardial infarction. AB - Animal models of sustained ischemia have shown exacerbation of myocardial injury during reperfusion, mediated largely by cytotoxic effects of free radical generation, complement activation, shifts in substrate use and inflammation. On the basis of current understanding of the pathogenesis of reperfusion injury, numerous therapies have shown a reduction in infarct size and improved ventricular function in animal models. Clinical trial experience has, however, so far been disappointing for the use of these therapies in patients with acute myocardial infarction (MI), being associated with no or inconsistent benefit to left-ventricular function, complications of MI or survival. The growing emphasis on early revascularization with primary coronary intervention and stenting in the management acute MI, with the potential for more complete drug delivery to injured myocardium, could provide greater opportunities for pharmacologic cardioprotection as adjunctive therapy in the future. PMID- 16265380 TI - Therapy insight: congestive heart failure, chronic kidney disease and anemia, the cardio-renal-anemia syndrome. AB - Congestive heart failure (CHF) and chronic kidney disease (CKD) often progress to end stage even with optimum medical therapy. One factor that is common to both conditions is anemia, which is present in about a third of CHF patients. CHF can cause or worsen both anemia and CKD, and CKD can cause or worsen both anemia and CHF. Thus, a vicious circle exists between these three conditions, with each causing or worsening the other. We have called this condition the cardio-renal anemia syndrome. Anemia in CHF is associated with increased mortality and hospitalization, reduced cardiac function and evidence of more severe CHF and CKD than in nonanemic patients. Intervention studies in anemic CHF patients have shown that optimum medical treatment of CHF and the correction of the associated anemia with subcutaneous erythropoietin and oral iron or intravenous iron sucrose can improve cardiac function, patients' functional status, renal function and quality of life, and reduce the frequency of hospitalization and the dose of diuretics required. PMID- 16265381 TI - Technology insight: transcoronary ablation of septal hypertrophy. AB - Among patients with hypertrophic cardiomyopathy, 5-10% have outflow tract obstruction that results in symptoms refractory to medical therapy. Surgical myectomy has been the gold standard for treating these patients since the 1960s. In the past 10 years, however, alcohol septal ablation has emerged as an alternative procedure. There is growing evidence to indicate that this technique is comparable to surgical myectomy with respect to hemodynamic and functional improvement, although no randomized comparative studies exist to date. The procedure consists of injecting as mall volume of alcohol into a septal perforator under balloon occlusion,preferably after localization of the target septal branch by myocardial contrast echocardiography. Relief of the gradient is instantaneous in about 90% of patients, and further decreases might occur over time. Functional class, exercise capacity and quality of life have been shown to be improved during follow-up. The main risk of the technique is atrioventricular block, which occurs in about 10% of patients. Because of this side-effect and a steep learning curve, the technique should be performed only by experienced operators and on carefully selected patients. PMID- 16265382 TI - Pulmonary artery sarcoma masquerading as chronic thromboembolic pulmonary hypertension. AB - BACKGROUND: A 40-year-old woman presented with dyspnea in the fifth month of pregnancy followed by a sudden onset of pleuritic chest pain 2 weeks postpartum. She was diagnosed as having pulmonary embolism by CT angiography and anticoagulated with heparin followed by warfarin. Despite 6 months of warfarin therapy she remained symptomatic. An echocardiogram demonstrated pulmonary hypertension, and a repeat CT angiogram demonstrated no change in the pulmonary artery filling defects. INVESTIGATIONS: Chest radiography, ventilation-perfusion scan, echocardiogram, right-heart catheterization, pulmonary angiography and pulmonary artery exploration. DIAGNOSIS: High-grade intimal sarcoma of the pulmonary artery. TREATMENT: Pulmonary endarterectomy. PMID- 16265383 TI - Novel potential perils of volunteering for clinical trials. PMID- 16265384 TI - Appendiceal function and dysfunction: what are the implications for inflammatory bowel disease? PMID- 16265398 TI - Can peptic ulcers be healed with 7 days of proton-pump-inhibitor-based triple therapy? PMID- 16265399 TI - Should azathioprine be withdrawn in patients with Crohn's disease who are in long term remission? PMID- 16265400 TI - How does donor age influence the success of liver transplantation in patients with hepatitis? PMID- 16265401 TI - Should African Americans be screened for colorectal cancer at an earlier age? PMID- 16265403 TI - Current management of locally advanced pancreatic cancer. AB - Almost 30% of patients with pancreatic cancer present with large, locally advanced tumors in the absence of distant metastases. Because surgical resection is frequently contraindicated by vascular invasion, locally advanced pancreatic cancer has a dismal prognosis with a 6-10-month median survival. Recent advances in the multimodality treatment of other gastrointestinal malignancies have not altered the management of patients with locally advanced pancreatic cancer, a clinical dilemma reflected by the number of nonrandomized trials and anecdotal reports addressing this difficult disease. Our review summarizes the current status of aggressive surgical resection and neoadjuvant chemoradiation for locally advanced pancreatic cancer and suggests a treatment algorithm for patients with this disease based upon published clinical evidence. PMID- 16265404 TI - Technology insight: Enteral stenting and new technology. AB - Self-expandable metal stents (SEMS) have gained acceptance for use in the gastrointestinal tract in order to relieve malignant luminal obstruction. In the upper gastrointestinal tract SEMS are used as an alternative to surgical bypass for palliation of malignant gastric-outlet obstruction. In the colon, SEMS are used to avoid colostomy during palliation and as a bridge to surgery for left sided colonic obstruction. Enteral SEMS appear to be cost effective. This article reviews the latest in stent technology as well as the outcomes following their placement. PMID- 16265405 TI - Supplemental evidence. PMID- 16265406 TI - Endoscopy in pregnancy: risks versus benefits. PMID- 16265407 TI - Mechanically restricted regional blood flow might explain gastrointestinal pain. PMID- 16265425 TI - Management of stress-related bleeding: evolution from the era before cimetidine. PMID- 16265426 TI - Aberrant crypt foci as biomarkers for colonic dysplasia. PMID- 16265427 TI - Molecular screening for Lynch syndrome. PMID- 16265428 TI - What is the best initial therapy for patients with uninvestigated heartburn? PMID- 16265429 TI - Is endoscopic variceal band ligation more effective than propranolol for the prevention of esophageal hemorrhage? PMID- 16265430 TI - Technology insight: liver support systems. AB - Emergency orthotopic liver transplantation (OLT) is currently the only standard treatment for fulminant hepatic failure (FHF). The waiting time for transplantation can exceed a week-using a liver assist device to bridge patients with FHF to OLT might therefore decrease the mortality rate. Several liver support systems have been described, but no system has gained FDA approval or widespread clinical acceptance. Although the results of many experimental and clinical trials are encouraging, the field is still in its initial stages. Using nonbiologic liver support is based on the assumption that several toxins that cause hepatic coma can be removed from the circulation by blood or plasma sorption methods. As these toxins could be involved in many FHF complications recovery without the need for transplantation is the ultimate aim. Biologic liver support uses xenogeneic livers or hepatocytes to support the failed human liver, exploiting biological cell functions, namely detoxification, metabolism, and biosynthesis. The classical nonbiologic dialysis methods could decrease mortality in patients with acute-on-chronic liver failure, but definitive conclusions are impossible to draw because of the small number of patients studied and inadequate follow-up. Larger studies performed in specialty centers should provide conclusive data about the role of the bioartificial liver support system as a possible universal bridge to OLT. This article presents an overview of published experience with liver support systems since the 1960s. PMID- 16265431 TI - Mechanisms of disease: defensins in gastrointestinal diseases. AB - Defensins are endogenous antibiotics with microbicidal activity against Gram negative and Gram-positive bacteria, fungi, viruses and protozoa. In the gastrointestinal tract, defensins help regulate the composition and number of colonizing microbes, and protect the host from food-borne and water-borne pathogens. In health, the normal host relationship with the commensal luminal microbiota is beneficial, but the same commensal bacteria could have a pathogenic role in inflammatory diseases. A disturbance in antimicrobial defense, as provided by Paneth cells of the small intestine, seems to be a critical factor in the pathogenesis of ileal Crohn's disease, an inflammatory disease of the intestinal tract. The disruption of the critical balance between antimicrobial peptides and luminal bacteria might also explain other gastrointestinal infections and diseases. Elucidating the underlying mechanisms involved in the regulation and biology of defensins could open up new therapeutic avenues. PMID- 16265432 TI - Mechanisms of disease: the role of intestinal barrier function in the pathogenesis of gastrointestinal autoimmune diseases. AB - The primary functions of the gastrointestinal tract have traditionally been perceived to be limited to the digestion and absorption of nutrients and electrolytes, and to water homeostasis. A more attentive analysis of the anatomic and functional arrangement of the gastrointestinal tract, however, suggests that another extremely important function of this organ is its ability to regulate the trafficking of macromolecules between the environment and the host through a barrier mechanism. Together with the gut-associated lymphoid tissue and the neuroendocrine network, the intestinal epithelial barrier, with its intercellular tight junctions, controls the equilibrium between tolerance and immunity to nonself-antigens. When the finely tuned trafficking of macromolecules is dysregulated in genetically susceptible individuals, both intestinal and extraintestinal autoimmune disorders can occur. This new paradigm subverts traditional theories underlying the development of autoimmunity, which are based on molecular mimicry and/or the bystander effect, and suggests that the autoimmune process can be arrested if the interplay between genes and environmental triggers is prevented by re-establishing intestinal barrier function. Understanding the role of the intestinal barrier in the pathogenesis of gastrointestinal disease is an area of translational research that encompasses many fields and is currently receiving a great deal of attention. This review is timely given the increased interest in the role of a 'leaky gut' in the pathogenesis of gastrointestinal diseases and the advent of novel treatment strategies, such as the use of probiotics. PMID- 16265434 TI - Childhood: a critical focus for 'primordial prevention' research. PMID- 16265433 TI - Mechanisms of disease: the genetic epidemiology of gallbladder stones. AB - Cholelithiasis is one of the most prevalent and most expensive gastroenterologic diseases. It belongs to the group of complex metabolic disorders that affect humans, and its critical pathogenic mechanisms are not well defined. As a result, primary or secondary prevention strategies are sparse, and the only effective treatment is cholecystectomy. Here we provide an update on the molecular pathogenesis of gallbladder stones, evidence supporting the hypothesis that genetic factors are key elements predisposing to gallstones, and progress in human genetic studies of cholesterol stones. Data from recent identical twin, family and linkage studies provide conclusive evidence for a strong genetic component to gallstone disease. Furthermore, epidemiologic studies in at-risk populations indicate that gallstone formation is caused by multiple environmental influences and common genetic factors and their interactions. By contrast, monogenic subtypes of cholelithiasis, such as ATP-binding-cassette transporter deficiencies, appear to be rare. The summary of human association studies illustrates that distinct common gene variants might contribute to gallstone formation in different ethnic groups. The characterization of lithogenic genes in knockout and transgenic mice and the identification of many gallstone susceptibility loci in inbred mice provide the basis for studies of the corresponding genes in patients with gallstones. The transfer of findings from mouse genetics to the bedside might lead to new strategies for individual risk assessment and reveal novel molecular targets for prevention and medical therapies. PMID- 16265435 TI - 2004 ACC/AHA guideline for the management of patients with STEMI: the implications for clinicians. PMID- 16265436 TI - The Fontan operation in clinical practice: indications and controversies. PMID- 16265450 TI - Should patients with stable coronary artery disease be treated with ACE inhibitor therapy? PMID- 16265451 TI - Do radial-artery grafts have a superior 1-year patency to saphenous-vein grafts? PMID- 16265452 TI - Val-HeFT: do angiotensin-receptor blockers benefit heart failure patients already receiving ACE inhibitor therapy? PMID- 16265453 TI - How common is rhabdomyolysis in patients receiving lipid-lowering therapy? PMID- 16265454 TI - Can recent-onset atrial fibrillation be safely and successfully treated with self administered antiarrhythmic drugs? PMID- 16265456 TI - Magnetic resonance angiography and the peripheral vasculature: how useful is it? PMID- 16265455 TI - Is obesity a risk factor for atrial fibrillation? PMID- 16265457 TI - Gene transfer for ischemic cardiovascular disease: is this the end of the beginning or the beginning of the end? AB - The past decade has represented a period of high expectations for cardiovascular gene transfer on the basis of the findings from preclinical experiments and promising early clinical results. Yet, randomized studies have not demonstrated similar results. Do these poor results mean that gene transfer for ischemic cardiovascular disease has failed in its promise, or do they merely signify the inherent challenges of a pioneering field? In this paper we briefly review the clinical experience of gene transfer for ischemic cardiovascular disease and propose future directions for research. PMID- 16265458 TI - Therapy Insight: interactions between atrial fibrillation and obstructive sleep apnea. AB - Atrial fibrillation (AF) and obesity are coinciding epidemics. Obstructive sleep apnea (OSA) correlates directly with obesity and is highly prevalent among middle aged adults. Emerging evidence supports a strong association between AF and OSA. The rate of AF among patients with OSA is low but is nevertheless higher than that in the general population. The prevalence of OSA among patients with AF is strikingly high-at least 32% and possibly as high as 49%-although differences in the AF populations studied and the criteria used to diagnose OSA have prevented more accurate measurement of this rate. Data in focused populations at risk of AF show that OSA increases the frequency and recurrence of AF. These findings cannot, however, be reliably generalized to the larger OSA population. Although the available information is limited, treatment of OSA, particularly with continuous positive airway pressure, seems to substantially reduce the frequency of AF. If true, this approach would provide a safe and noninvasive therapy for some patients with AF. Randomized controlled trials are necessary to rigorously answer this question. Many data have been obtained from studies with focuses other than the direct relationship between OSA and AF. Studies carefully designed to investigate the relevant issues are now needed. PMID- 16265459 TI - Technology insight: assessment of myocardial viability by delayed-enhancement magnetic resonance imaging. AB - Myocardial viability is of established importance to the management of cardiac patients being considered for revascularization. Existing noninvasive imaging tests to examine myocardial viability, such as stress echocardiography and nuclear scintigraphy, are of recognized utility but are subject to intrinsic limitations. Over the past few years delayed-enhancement MRI (DE-MRI) has emerged as an alternative to traditional tests and for the first time allows direct visualization of the transmural extent of myocardial viability. In this paper we review the scientific data that underlie the use of DE-MRI in patients with ischemic heart disease. Progress in this area is largely the result of the development of a new MRI pulse sequence in the late 1990s, which improved the detection of necrotic and scarred myocardial tissue. Following this technical development, a series of detailed histologic comparisons in large animal models revealed that both acute and healed myocardial infarcts appeared as brighter (hyperenhanced) areas than viable regions, and that the effect is independent of contractile function. The resulting 'bright is dead' hypothesis has thus far proven of significant use in patients with ischemic heart disease. Data are now emerging which suggest that the DE-MRI technique also has important implications for patients with nonischemic forms of cardiomyopathy. PMID- 16265460 TI - Technology insight: catheter ablation of the pulmonary veins in the treatment of atrial fibrillation. AB - Over the past 5 years, catheter ablation of atrial fibrillation (AF) has evolved from being an experimental procedure to one that is performed in most large medical centers throughout the world. The rapid and widespread acceptance of this procedure reflects encouraging reports of the safety and efficacy of a pulmonary vein (PV) approach, which has 70-80% efficacy and around a 5% risk of major complications. Currently three PV-based ablation strategies are employed for catheter ablation of AF, termed segmental PV isolation, circumferential PV ablation and circumferential PV isolation. Three main research activities are ongoing in the field of catheter ablation of AF. One aim is to better define the safety and efficacy of AF ablation using the three approaches listed. The second research focus is to improve safety and efficacy by the development of new mapping and ablation tools. And the third major area of research is the development of new strategies for AF ablation. It is highly likely during the next 5 years that catheter ablation of AF will become a routine and well-accepted treatment strategy for this common arrhythmia condition. PMID- 16265462 TI - Chrysotile asbestos exposure associated with removal of automobile exhaust systems (ca. 1945-1975) by mechanics: results of a simulation study. AB - For decades, asbestos-containing gaskets were used in virtually every system that involved the transport of fluids or gases. Prior to the mid-1970s, some automobile exhaust systems contained asbestos gaskets either at flanges along the exhaust pipes or at the exhaust manifolds of the engine. A limited number of automobile mufflers were lined with asbestos paper. This paper describes a simulation study that characterized personal and bystander exposures to asbestos during the removal of automobile exhaust systems (ca. 1945-1975) containing asbestos gaskets. A total of 16 pre-1974 vehicles with old or original exhaust systems were studied. Of the 16 vehicles, 12 contained asbestos gaskets in the exhaust system and two vehicles had asbestos lining inside the muffler. A total of 82 samples (23 personal, 38 bystander, and 21 indoor background) were analyzed by Phase Contrast Microscopy (PCM) and 88 samples (25 personal, 41 bystander, and 22 indoor background) by Transmission Electron Microscopy (TEM). Only seven of 25 worker samples analyzed by TEM detected asbestos fibers and 18 were below the analytical sensitivity limit (mean 0.013 f/cc, range 0.001-0.074 f/cc). Applying the ratio of asbestos fibers:total fibers (including non-asbestos) as determined by TEM to the PCM results showed an average (1 h) adjusted PCM worker exposure of 0.018 f/cc (0.002-0.04 f/cc). The average (1 h) adjusted PCM airborne concentration for bystanders was 0.008 f/cc (range 0.0008-0.015 f/cc). Assuming a mechanic can replace four automobile single exhaust systems in 1 workday, the estimated 8-h time-weighted average (TWA) for a mechanic performing this work was 0.01 f/cc. Under a scenario where a mechanic might repeatedly conduct exhaust work, these results suggest that exposures to asbestos from work with automobile exhaust systems during the 1950s through the 1970s containing asbestos gaskets were substantially below 0.1 f/cc, the current PEL for chrysotile asbestos, and quite often were not detectable. PMID- 16265461 TI - A rare case of acute 'infective' myocardial infarction triggered by acute parvovirus B19 myocarditis. AB - BACKGROUND: A 25-year-old obese male (BMI 31.9 kg/m(2)) presented with atypical chest pain of sudden onset that was indistinguishable from acute myocardial infarction. He had tachycardia (104 beats/min) and dyspnea at a low level of exercise. He had no previous cardiac history, but his cardiovascular risk profile included a familial predisposition, smoking and hypertension. INVESTIGATIONS: Electrocardiogram, laboratory testing, chest radiography, echocardiography, coronary angiography, intravascular ultrasonography and endomyocardial biopsy. DIAGNOSIS: Acute myocardial infarction and parvovirus-B19-positive myocarditis. MANAGEMENT: Percutaneous transluminal coronary angioplasty with intracoronary abciximab, heparin and nitroglycerin infusion. PMID- 16265463 TI - Treatment of acute coronary syndromes: have we reached the ceiling? PMID- 16265464 TI - Over-the-counter statins: a new controversy. PMID- 16265477 TI - Off-pump versus on-pump CABG surgery: how do the long-term graft patencies compare? PMID- 16265478 TI - Is rescue reperfusion beneficial after failed thrombolysis in patients with acute myocardial infarction? PMID- 16265479 TI - Can nicorandil treat angina pectoris effectively? PMID- 16265480 TI - Should every patient with left-ventricular systolic dysfunction receive implantable cardioverter-defibrillators? PMID- 16265481 TI - Does coronary artery revascularization before major vascular surgery benefit patients with coronary artery disease? PMID- 16265482 TI - Do implantable cardioverter-defibrillators benefit patients immediately after acute myocardial infarction? PMID- 16265483 TI - The prognostic value of N-terminal proB-type natriuretic peptide. AB - The heart is not only a pump, but also it is an endocrine organ. Cardiac stretch and overload stimulate the secretion of natriuretic peptides, which have a variety of beneficial actions, such as vasodilation and natriuresis. Cardiac derived natriuretic peptides, especially B-type natriuretic peptide (BNP), have emerged as useful biomarkers for the diagnosis, and potentially the treatment, of heart failure patients. The inactive amino-terminal fragment of the BNP prohormone (NT-proBNP), which is more stable than mature BNP, has also been recognized as an aid in the diagnosis of left-ventricular systolic dysfunction. Furthermore, elevated NT-proBNP concentrations have been shown to be predictive of poor prognosis in a variety of cardiovascular diseases, suggesting that it could be useful for risk stratification of patients. This review summarizes current literature that has addressed the issue of NT-proBNP as a prognostic tool in heart failure, acute coronary syndromes and other conditions. PMID- 16265484 TI - The Fontan circulation: a challenge to William Harvey? AB - The Fontan operation, which places the systemic and pulmonary circulations in series and is driven by a single ventricular chamber, is the treatment of choice for patients born with one ventricle. Its introduction 35 years ago was the result of a flurry of experimental and clinical research that had started in the 1940s. A large number of children have benefited and continue to benefit from the Fontan operation, but there is a genuine concern that, despite the refinement of the surgical procedures in the past 20 years, continuing attrition might be inevitable. This adverse effect can lead to a decline in functional capacity, and premature late death. PMID- 16265485 TI - Mechanisms of disease: pathologic structural remodeling is more than adaptive hypertrophy in hypertensive heart disease. AB - Changes in the composition of cardiac tissue develop in arterial hypertension and lead to structural remodeling of the myocardium. Structural remodeling is the consequence of a number of pathologic processes, mediated by mechanical, neurohormonal and cytokine routes, occurring in the cardiomyocyte and the noncardiomyocyte compartments of the heart. One of these processes is related to the disruption of the equilibrium between the synthesis and degradation of collagen type I and III molecules, which results in an excessive accumulation of collagen type I and III fibers in the interstitium and the perivascular regions of the myocardium. The clinical relevance of ventricular fibrosis is that it might contribute to the increased cardiac risk of patients with hypertensive heart disease. This review focuses on the mechanisms of hypertensive ventricular fibrosis and its clinical consequences. In addition, we discuss the noninvasive methods for the diagnosis of cardiac fibrosis and the therapeutic strategies aimed to promote its reduction. PMID- 16265486 TI - Technology insight: hand-carried ultrasound cardiac assessment--evolution, not revolution. AB - Hand-carried ultrasound devices can enhance the art of bedside physical examination by increasing diagnostic accuracy, detecting disease at an earlier stage, and improving triage and referral of patients. Although limitations of device cost and portability can be overcome with technologic advances, the shortage of standardized imaging and training opportunities now needs to be addressed to move the technique forward. Cardiologists are the best-qualified subspecialists to design and teach a simplified training program for bedside use of hand-carried ultrasound devices to assess the cardiovascular system. PMID- 16265487 TI - Atrial fibrillation: an epidemiologic, scientific and clinical challenge. PMID- 16265488 TI - Anticoagulants for stroke prevention in atrial fibrillation: what is the right rate and where do we go from here? PMID- 16265502 TI - Can niacin slow the development of atherosclerosis in coronary artery disease patients already taking statins? PMID- 16265503 TI - Does losartan reduce the risk of stroke in patients with hypertension? PMID- 16265504 TI - Can the measurement of impaired flow-mediated dilation predict in-stent restenosis? PMID- 16265505 TI - Is self-management of oral anticoagulation a feasible and safe option? PMID- 16265506 TI - Can intravascular ultrasound detect left main coronary artery disease accurately? PMID- 16265507 TI - Does reducing a patient's CRP levels with statin therapy reduce their risk of recurrent cardiovascular events? PMID- 16265508 TI - Atherosclerosis of the ascending aorta as a major determinant of the outcome of cardiac surgery. AB - Epiaortic ultrasonography has high sensitivity for the detection of atherosclerosis. In several studies, the technique has identified atherosclerosis of the ascending aorta as the major risk factor for stroke after cardiac surgery. The level of risk depends on the presence, location and extent of disease when the ascending aorta is surgically manipulated. This knowledge enables clinicians to focus on the diagnostic and surgical technique and to consider the various options. Routine use of intraoperative epiaortic ultrasonography should be applied so that surgical manipulation of the ascending aorta can be reduced or, if possible, avoided in patients with atherosclerosis of the ascending aorta. Alternatively, if major manipulation such as clamping must be performed in the presence of severe atherosclerosis, the use of intra-aortic filters could be considered. PMID- 16265509 TI - Therapy insight: heart disease and the insulin-resistant patient. AB - Insulin-resistance syndromes are of pandemic proportions; 150 million people worldwide and an estimated 43 million people in the US are currently affected by type 2 diabetes mellitus or metabolic syndrome respectively. Treatment of heart disease in the context of type 2 diabetes requires multifactorial risk-factor management, including lifestyle modification and drug treatment for comorbidities. Management of coronary risk extends beyond simple cholesterol lowering. Early use of cardiac imaging and, where appropriate, revascularization should be considered in high-risk or symptomatic patients. Traditionally, patients with type 2 diabetes and coronary arterial disease have been treated surgically, but percutaneous revascularization of these patients is increasingly common. Indeed, revascularization by use of drug-eluting coronary stents combined with administration of novel antiplatelet agents has revolutionized percutaneous coronary intervention in patients with type 2 diabetes. Despite these advances, there is no consensus of opinion regarding revascularization strategies or risk factor management in insulin-resistant patients with symptomatic or prognostically important coronary arterial disease. Furthermore, specific therapies and preventative strategies for diabetic cardiomyopathy and heart failure in patients with type 2 diabetes remain elusive. The identification of optimized approaches for the prevention and treatment of the metabolic syndrome and heart disease in insulin-resistant, nondiabetic patients remains a major global challenge. PMID- 16265510 TI - Technology insight: randomized trials of off-pump versus on-pump coronary artery bypass surgery. AB - Coronary artery bypass grafting has proven a remarkably effective treatment for occlusive coronary artery disease, with demonstrable impact on both symptoms and survival. As conducted traditionally, cardiopulmonary bypass is required, and a global myocardial ischemic insult imposed with aortic occlusion under the protection of cardioplegic arrest. Despite the remarkable success of this approach, concerns over the systemic effects of bypass, including neurologic sequelae as well as ischemic myocardial injury, have stimulated development of techniques and technology to perform coronary bypass 'off-pump'. This technique obviates the need for the bypass machine and imposes only brief regional ischemia during construction of each individual anastomosis. Despite enthusiastic support by a devoted cohort of surgeons, and a host of nonrandomized retrospective studies demonstrating an apparent benefit to the off-pump technique, the technique has not been universally adopted. How can there be such controversy over what appears to be a superior approach? In part, many surgeons are concerned that the greater technical difficulty of the technique will impact long-term results adversely. There is also uncertainty with regard to the actual advantage of off-pump coronary artery bypass over the tried-and-true methods. Surgeons recognize that the results of any surgical series are particularly subject to the influence of subtle selection biases. Accordingly, prospective randomized studies add particular value to the debate. It is the aim of this review to examine the evidence for off-pump coronary artery bypass critically, from a surgeon's perspective, with particular emphasis on knowledge derived from a representative selection of published prospective randomized studies. PMID- 16265511 TI - Guidelines on hypertension in 2005: a commentary. PMID- 16265512 TI - Informed consent: theory versus practice. PMID- 16265528 TI - Can creatine kinase MB levels predict mortality after cardiac surgery? PMID- 16265527 TI - Should ximelagatran replace warfarin for stroke prevention in patients with atrial fibrillation? PMID- 16265529 TI - Fractional flow reserve: can it predict adverse events accurately after coronary stenting? PMID- 16265530 TI - Should patients with combined hyperlipidemia receive statin-fibrate combination therapy? PMID- 16265531 TI - Can bedside testing of B-type natriuretic peptide be used to diagnose left ventricular diastolic dysfunction? PMID- 16265532 TI - Cardiac resynchronization therapy: does varying the pacing site or combination of sites improve cardiac function? PMID- 16265534 TI - Therapy insight: cardiovascular complications associated with muscular dystrophies. AB - The muscular dystrophies are commonly associated with cardiovascular complications, including cardiomyopathy and cardiac arrhythmias. These complications are caused by intrinsic defects in cardiomyocyte and cardiac conduction system function, and by the presence of severe skeletal muscle disease, which also contributes to cardiac dysfunction. Unlike the skeletal muscle degenerative process, for which treatment options are currently limited, therapy is available for the cardiovascular complications that accompany muscular dystrophy. New therapies for skeletal muscle degeneration are moving into clinical trials and, ultimately, into clinical practice. These therapies are expected to also improve the cardiac function, longevity and wellbeing of muscular dystrophy patients. PMID- 16265533 TI - Drug insight: cyclo-oxygenase 2 inhibitors and cardiovascular risk--where are we now? AB - Cyclo-oxygenase (COX) 1 mediates the production of thromboxane A2 in platelets, leading to platelet aggregation and vasoconstriction. Conversely, COX2 catalyzes endothelial prostacyclin synthesis, which effectively counteracts thromboxane A2, triggering vasodilation and platelet inhibition. Selective COX2 inhibitors decrease prostacyclin production, potentially disrupting homeostasis and creating a prothrombotic state. The VIGOR study findings of increased cardiovascular risk with rofecoxib were subsequently confirmed by large meta-analyses, observational studies and recent APPROVe trial publication. The APC trial findings of increased cardiovascular risk with Celebrex (celecoxib) conflict with those in the ADAPT trial, the upcoming PreSAP publication, a case-control study by Graham et al. and prior large clinical trials, meta-analyses and observational studies of this drug. Therefore, while an adverse class effect is a possibility for COX2 inhibitors, the published data are inconsistent. Baseline cardiovascular risk in patients might contribute significantly to these findings. In light of the negative Vioxx (rofecoxib) publicity, however, COX2 inhibitors might forever remain underinvestigated. The relative selectivity of these compounds for COX2 is extremely variable, casting significant doubt on the class-effect hypothesis. Improved endothelial function has also been reported with celecoxib, leading to endothelium-dependent vasodilation, and associated decreases in C-reactive protein and LDL cholesterol. The addition of meloxicam to low-dose aspirin and heparin has improved clinical outcomes after acute coronary syndromes. These are the first studies suggesting improvement in endothelial function and reduction of inflammation with COX2 inhibition. Thus, more randomized controlled trials are needed to study the relative cardiovascular effects of different COX2 inhibitors, alone and in combination with aspirin. PMID- 16265535 TI - Mechanisms of disease: macrophage-derived foam cells emerging as therapeutic targets in atherosclerosis. AB - The limited efficacy of current treatment strategies for targeting atherosclerosis and its complications requires new therapeutic options to be explored. From early fatty-streak lesions to advanced plaques, macrophage-derived foam cells are integral to the development and progression of atherosclerosis. Elucidation of molecular and cellular processes involving macrophages has led to numerous therapeutic targets being suggested. Potential sites of intervention range from monocyte recruitment, through cholesterol uptake and esterification, to cholesterol evacuation and macrophage egress from plaque. In addition, complex patterns of transcriptional regulation of genes involved in macrophage lipid homeostasis and in the regulation of inflammation have been partly unraveled. Recognition of ATP-binding cassette cholesterol transport mechanisms and cellular interactions with cholesterol-accepting apolipoproteins (or synthetic mimetics) opens up new potential therapies to induce atherosclerosis regression in humans. This review presents a systematic evaluation of actual and potential macrophage directed pharmacologic interventions. It reflects the timely convergence of three important strands: advances in molecular and cell biology that have suggested therapeutic targets in macrophages; the development of multiple classes of drugs targeting these pathways; and the emergence of sensitive imaging techniques that have enabled identification of changes in plaque size and composition in response to treatment. PMID- 16265537 TI - The evolving role of CT and MRI in atherothrombotic evaluation and management. PMID- 16265536 TI - Staged endovascular treatment for complicated type B aortic dissection. AB - BACKGROUND: A 40-year-old man presented with acute chest and back pain, hypertension and anuria. Two years previously he had been diagnosed with acute uncomplicated type B aortic dissection. Following conservative management, with aggressive antihypertensive therapy and analgesia, he was monitored with 6 monthly surveillance CT scans. These demonstrated a complicated type B dissection with renal and iliac malperfusion. INVESTIGATIONS: Multislice CT, transthoracic and transesophageal echocardiography, digital subtraction aortography. DIAGNOSIS: Acute-on-chronic type B aortic dissection, complicated by aneurysmal dilatation of the thoracic aorta and visceral malperfusion. MANAGEMENT: Antihypertensive therapy; staged thoracoabdominal and branch vessel endoluminal repair (STABLE procedure), with stabilization of the dissection and rescue of renal function; CT imaging surveillance to monitor for any further complications. PMID- 16265538 TI - Hypertrophic cardiomyopathy in the community: why we should care. PMID- 16265553 TI - Can losartan prevent new-onset atrial fibrillation in hypertensive patients more effectively than atenolol? PMID- 16265554 TI - Could risk stratification aid the treatment of patients with acute decompensated heart failure? PMID- 16265555 TI - Does quantitative assessment of asymptomatic mitral regurgitation predict patients' outcomes? PMID- 16265556 TI - Does carvedilol have antiarrhythmic properties? PMID- 16265557 TI - Is balloon aortic valvuloplasty a feasible treatment for neonates and young infants with aortic valve stenosis? PMID- 16265558 TI - Can the therapeutic window for tissue plasminogen activator be extended to 6 hours after stroke onset? PMID- 16265559 TI - Could clopidogrel's platelet inhibition be enhanced by an increased loading dose and eptifibatide? PMID- 16265560 TI - Management of long QT syndrome. AB - Congenital long QT syndrome (LQTS) is a genetic disorder characterized by prolongation of the QT interval on the electrocardiogram and by life-threatening cardiac arrhythmias, occurring especially during conditions of increased sympathetic activity. Existing therapies are very effective, but mortality is high among untreated, symptomatic individuals. The identification of several of the genes responsible for LQTS and the realization that they all encode cardiac ion-channels has represented a landmark finding. This advance has fostered novel genotype-phenotype studies that are providing unique insight into how close the relationship can be between molecular biology and clinical cardiology. LQTS represents a paradigm for sudden cardiac death. Indeed, the growing knowledge developed for LQTS is likely to provide the key to understanding the genetic propensity to sudden death in patients with more-common cardiovascular diseases. The data presented here illustrate how the treatment of LQTS is rapidly evolving toward a highly individually tailored approach on the basis of patient-specific genetic information. PMID- 16265561 TI - Sudden cardiac death: mechanisms, therapies and challenges. AB - Sudden cardiac death (SCD) is the leading cause of cardiac death in the US. In the past few years, intensive efforts have been made to expand public awareness of SCD and to increase our understanding of its pathophysiology, medical treatment options and device therapy. Significant advances have been made in our ability to prevent SCD in both primary and secondary health care. Two critical issues remain, however: the identification of patients who would benefit from such therapies, and how to achieve even greater prevention, especially primary prevention. The goal of this article is to provide a review of the topic of SCD in the setting of abnormal myocardial substrate, to outline the techniques that are useful in identifying patients at risk, and available treatment options. PMID- 16265562 TI - Technology insight: possible applications of multislice computed tomography in clinical cardiology. AB - With the introduction of four-slice scanners in 1999, multislice CT (MSCT) technology became available for investigative examination of the heart. Since then, MSCT technology has undergone rapid technical progress; temporal and spatial resolutions have been especially improved. The improved diagnostic image quality has led to more possible uses of MSCT being defined. At present, issues such as visualization of coronary artery bypass grafts, detection of stenoses of native coronary arteries, description of coronary anomalies, and calcium scoring, can be investigated reasonably well. Other features, such as plaque imaging and visualization of intracoronary stents, need further evaluation. A large number of factors, however, such as heart rate, atrial fibrillation, breathing artefacts and severe calcification, still influence image quality and reduce validity. In this article we provide a summary of current fields of application of cardiac MSCT. The word 'indication' is consciously avoided because official guidelines for the use of MSCT in heart examination have not yet been issued. Hopefully, prospective multicenter trials will be performed soon, providing more data with which to establish guidelines for both cardiologist and radiologist. PMID- 16265563 TI - Intracardiac leiomyomatosis: iliac vein to right-ventricular outflow tract. AB - BACKGROUND: A 58-year-old female with a history of hypertension and asthma presented to an internist for a routine physical examination. A grade II/VI systolic ejection murmur and electrocardiogram abnormalities were noted. She was referred to a cardiologist for further assessment. INVESTIGATIONS: Transthoracic echocardiography, transesophageal echocardiography, contrast-enhanced CT and MRI, exploratory laparotomy. DIAGNOSIS: Intracardiac leiomyomatosis. MANAGEMENT: Surgical excision. PMID- 16265564 TI - Atherothrombosis as a systemic, often silent, disease. PMID- 16265565 TI - Childhood obesity and cardiovascular disease: the challenge ahead. PMID- 16265580 TI - Is eplerenone a cost-effective therapy for heart failure patients after myocardial infarction? PMID- 16265581 TI - Cardiac dimensions and function in atrial fibrillation: how do rate and rhythm control compare? PMID- 16265582 TI - Is clopidogrel plus aspirin more cost-effective than aspirin alone in high-risk patients with acute coronary syndromes? PMID- 16265583 TI - Pulmonary-vein cryoisolation versus left-atrial linear cryoablation for atrial fibrillation. PMID- 16265584 TI - Does angiotensin-converting-enzyme inhibitor therapy improve cognitive function in heart failure patients? PMID- 16265585 TI - Large-artery stiffness, hypertension and cardiovascular risk in older patients. AB - Research studies on hypertension have, so far, generally focused on vascular resistance and small arteries. The high prevalence of systolic hypertension in patients older than 50 years and the development of noninvasive Doppler and echotracking techniques have made it possible to determine large-artery stiffness with a high degree of reproducibility. Increased arterial stiffness and disturbed wave reflections are the basis for understanding reduced aortic elasticity and systolic hypertension, particularly in older people. This hemodynamic pattern results from mechanical factors and other pressure-independent risk factors, such as diabetes mellitus, renal failure, obesity and severe atherosclerosis. Distinct phenotypes of arterial stiffness and pulse pressure result from specific gene polymorphisms, such as those related to the renin-angiotensin system. The roles of arterial stiffness and wave reflections in hypertension have been elucidated by modern interpretations of the blood-pressure curve in relation to its propagation, mechanisms of systolic-blood-pressure amplification, and the pulse pressure amplitude. New predictors of cardiovascular risk have been identified, such as increased pulse pressure and pulse-wave velocity, and disturbed wave reflections, all of which are independent predictors of cardiovascular risk that are more powerful than either systolic or diastolic blood pressure alone. Therapeutic trials are investigating ways to reduce stiffness, and thereby allow the selective reduction of systolic and pulse pressure in hypertensive patients with or without advanced renal failure. Modern pharmacologic agents need to be identified, which could reduce systolic hypertension in patients older than 50 years. Here we discuss the structural and functional factors that influence arterial stiffness, wave reflections and pulse pressure in hypertension, as well as their related roles in cardiovascular risk. PMID- 16265586 TI - The role of shear stress in the destabilization of vulnerable plaques and related therapeutic implications. AB - American Heart Association type IV plaques consist of a lipid core covered by a fibrous cap, and develop at locations of eccentric low shear stress. Vascular remodeling initially preserves the lumen diameter while maintaining the low shear stress conditions that encourage plaque growth. When these plaques eventually start to intrude into the lumen, the shear stress in the area surrounding the plaque changes substantially, increasing tensile stress at the plaque shoulders and exacerbating fissuring and thrombosis. Local biologic effects induced by high shear stress can destabilize the cap, particularly on its upstream side, and turn it into a rupture-prone, vulnerable plaque. Tensile stress is the ultimate mechanical factor that precipitates rupture and atherothrombotic complications. The shear-stress-oriented view of plaque rupture has important therapeutic implications. In this review, we discuss the varying mechanobiologic mechanisms in the areas surrounding the plaque that might explain the otherwise paradoxical observations and unexpected outcomes of experimental therapies. PMID- 16265587 TI - Drug insight: bleeding after percutaneous coronary intervention-risks, measures and impact of anticoagulant treatment options. AB - In Europe, the use of interventional cardiology, including percutaneous coronary intervention (PCI), is increasing rapidly. The use of anticoagulation agents in PCI procedures is essential, but despite technical improvements, a significant associated bleeding risk still exists: more than 5% of patients are estimated to require transfusion, and around a further 13% experience minor bleeding. The methods used to detect and measure blood loss following PCI, however, vary widely between institutions and clinical trials. The risk of bleeding is influenced by therapeutic options and patient-specific characteristics, such as age, anemia and previous exposure to anticoagulants. Bleeding is associated with death, and also with less severe conditions such as thrombocytopenia, anemia, and hematoma, which have major impacts on patients' welfare and length of hospital stay, and on hospital budgets. Unfractionated heparin is the most widely used anticoagulant during PCI. Heparin, antiplatelet agents and other anticoagulants, however, have limitations that make it difficult to achieve a level of anticoagulation that prevents ischemic events without promoting bleeding. The use of low-molecular weight heparin and the addition of glycoprotein IIb/IIIa inhibitors offer improved outcomes, but safer and more effective therapeutic agents are still required. New anticoagulants, including direct thrombin inhibitors such as bivalirudin, show similar levels of efficacy to heparin plus glycoprotein IIb/IIIa inhibitors, but with fewer hemorrhagic complications, and might advance clinical practice. This review evaluates the impact of PCI-related bleeding on patients' outcomes and hospital resources, examines methods for the detection and measurement of bleeding, and appraises the therapeutic options--particularly the newer agents--available to minimize hemorrhagic complications. PMID- 16265588 TI - Mechanisms of disease: beta-adrenergic receptors--alterations in signal transduction and pharmacogenomics in heart failure. AB - Beta-adrenergic signaling is an important regulator of myocardial function. During the progression of heart failure (HF), a reproducible series of biochemical events occurs that affects beta-adrenergic receptor (beta-AR) signaling and cardiac function. Furthermore, there are pathophysiologic alterations in the expression and regulation of proteins that are regulated by beta-ARs during HF. Analyses of these complex signaling pathways have led to a better understanding of HF mechanisms and the use of beta-adrenergic antagonists, which have notably altered HF-related morbidity and mortality. Despite therapeutic advances that have affected beta-AR signaling, HF remains a leading cause of hospitalization and a principal cause of death in industrialized nations. In this review, we summarize current insights into beta-adrenergic signal-transduction pathways, the best-described beta-AR polymorphisms, and therapies that target the beta-AR pathway in HF. PMID- 16265589 TI - Vasospastic angina pectoris associated with Churg-Strauss syndrome. AB - BACKGROUND: A 50-year-old woman presented with recurrent episodes of unstable angina pectoris refractory to vasodilator treatment. Relevant coronary stenoses were excluded by coronary angiography and intravascular ultrasonography. Intracoronary infusion of acetylcholine revealed diffuse coronary vasospasms associated with clinical signs of myocardial ischemia and ST-segment elevation. Symptoms of bronchial asthma, polyneuropathy, nasal polyps, allergic rhinitis, gastritis and eosinophilia led to a diagnosis of Churg-Strauss syndrome. INVESTIGATIONS: Serum chemistry, coronary angiography, left-heart catheterization, intravascular ultrasonography and coronary vasospasm provocation with acetylcholine. DIAGNOSIS: Vasospastic angina pectoris associated with Churg Strauss syndrome. MANAGEMENT: Treatment with systemic corticosteroids and cyclophosphamide. PMID- 16265590 TI - Kinobeon A, purified from cultured safflower cells, is a novel and potent singlet oxygen quencher. AB - We recently reported that kinobeon A, produced from safflower cells, suppressed the free radical-induced damage of cell and microsomal membranes. In the present study, we investigated whether kinobeon A quenches singlet oxygen, another important active oxygen species. Kinobeon A inhibited the singlet oxygen-induced oxidation of squalene. The second-order rate constant between singlet oxygen and kinobeon A was 1.15 x 10(10) M(-1)s(-1) in methanol containing 10% dimethyl sulfoxide at 37 degrees C. Those of alpha-tocopherol and beta-carotene, which are known potent singlet oxygen quenchers, were 4.45 x 10(8) M(-1)s(-1) and 1.26 x 10(10) M(-1)s(-1), respectively. When kinobeon A was incubated with a thermolytic singlet oxygen generator, its concentration decreased. However, this change was extremely small compared to the amount of singlet oxygen formed and the inhibitory effect of kinobeon A on squalene oxidation by singlet oxygen. In conclusion, kinobeon A was a strong singlet oxygen quencher. It reacted chemically with singlet oxygen, but it was physical quenching that was mainly responsible for the elimination of singlet oxygen by kinobeon A. Kinobeon A is expected to have a preventive effect on singlet oxygen-related diseases of the skin or eyes. PMID- 16265591 TI - The effect of new non-cross resistant antitumour agents on the energy state of human erythrocytes. AB - Multidrug resistance (MDR) of tumour cells is related to the overexpression of ATP-dependent pumps responsible for the active efflux of antitumour agents out of resistant cells. Benzoperimidine and anthrapyridone compounds exhibit comparable cytotoxic activity against sensitive and MDR tumour cells. They diffuse extremely rapidly across the plasma membrane and render the ATP-dependent efflux inefficient. Such uptake could disturb an energy metabolism of normal cells possessing an elevated level of ATP-dependent proteins, especially erythrocytes having a high level of the MRP1, MRP4 and MRP5 proteins. In this study the effect of five antitumour agents: benzoperimidine (BP1), anthrapyridones (CO1, CO7) and reference drugs used in the clinic: doxorubicin (DOX) and pirarubicin (PIRA), on the energetic state in human erythrocytes has been examined. These compounds have various types of structure and kinetics of cellular uptake (slow--DOX, CO7, moderate--PIRA, fast--BP1, CO1) resulting in their different ability to saturate ATP-dependent transporters. The energetic state of erythrocytes was examined by determination of purine nucleotide contents (ATP, ADP, AMP), NAD(+) and values of adenylate energy charge (AEC) using an HPLC method. It was found that the level of nucleotides as well as the AEC value of erythrocytes were not changed during 24 h of incubation with these agents independently of their structure and ability to saturate ATP-dependent pumps. This is a very promising result in view of their potential use in the clinic as antitumour drugs against multidrug resistant cancers. PMID- 16265592 TI - Recent advances in studies on biochemical and structural properties of equilibrative and concentrative nucleoside transporters. AB - Nucleoside transporters (NT) facilitate the movement of nucleosides and nucleobases across cell membranes. NT-mediated transport is vital for the synthesis of nucleic acids in cells that lack de novo purine synthesis. Some nucleosides display biological activity and act as signalling molecules. For example, adenosine exerts a potent action on many physiological processes including vasodilatation, hormone and neurotransmitter release, platelet aggregation, and lipolysis. Therefore, carrier-mediated transport of this nucleoside plays an important role in modulating cell function, because the efficiency of the transport processes determines adenosine availability to its receptors or to metabolizing enzymes. Nucleoside transporters are also key elements in anticancer and antiviral therapy with the use of nucleoside analogues. Mammalian cells possess two major nucleoside transporter families: equilibrative (ENT) and concentrative (CNT) Na(+)-dependent ones. This review characterizes gene loci, substrate specificity, tissue distribution, membrane topology and structure of ENT and CNT proteins. Regulation of nucleoside transporters by various factors is also presented. PMID- 16265593 TI - Different properties of four molecular forms of protein kinase CK2 from Saccharomyces cerevisiae. AB - CK2 is a pleiotropic constitutively active serine/threonine protein kinase composed of two catalytic alpha- and two regulatory beta-subunits, whose regulation is still not well understood. It seems to play an essential role in regulation of many cellular processes. Four active forms of CK2, composed of alphaalpha'betabeta', alpha(2)betabeta', alpha'(2)betabeta', and a free alpha' subunit were isolated from wild-type yeast and strains containing a single deletion of the catalytic subunit. Each species exhibits properties typical for CK2, but they differ in substrate specificity and sensitivity to inhibitors. This suggests that each CK2 isomer may regulate different process or may differ in the way of its regulation. PMID- 16265594 TI - The role of benzoate secreted by Desulfotomaculum acetoxidans DSM 771 in sulfate uptake. AB - This work was designed to find the cause of the delay in hydrogen sulfide dissimilation in Desulfotomaculum acetoxidans DSM 771, which is dependent on the sulfate uptake. This bacterium grown without addition of any aromatic compound was shown by spectrum analysis with the methylene method to contain hydroxy benzoate derivatives. The presence of these compounds was confirmed by HPLC in fractions obtained from cell walls after 15 days of culture. The test with 2,2' azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt seemed to indicate the presence of peroxidase, which probably oxidized benzoate to its hydroxy derivatives. The test with 5-sulfo-salicylic acid proved the ability of the investigated strain to utilize arylsulfates and to reduce sulfate group to hydrogen sulfide. On the basis of the above data, we propose the following sequence of reactions: 1, benzoate secretion; 2, benzoate hydroxylation; 3, sulfonation of hydroxy-benzoate derivatives. PMID- 16265595 TI - Osteopontin gene expression in the aorta and the heart of propylthiouracil induced hypothyroid mice. AB - It is known that there is abnormal osteopontin (OPN) expression at the sites of atherosclerotic lesions. In the Apolipoprotein E gene knockout (ApoE-KO) mouse, a model of the atherosclerotic process, altered cholesterol metabolism with associated increase in OPN expression is evident at 12-22 weeks in the aorta and at 22 weeks in the heart. In this study, we analyzed another animal model of hypothyroid mice created by ingestion of propylthiouracil (PTU). After 2 weeks of PTU ingestion, the animals had significant decreases in thyroid hormones (T3 and T4) and immediate increases in blood lipids/cholesterol. Hypothyroid mice showed 1.3-, 1.5-, 2-fold increases in blood levels of total cholesterol, triglycerides, and low density lipoprotein-cholesterol respectively. Semi-quantitative RT-PCR analysis showed that hypothyroid mice had 1.4- to 2-fold increases of OPN mRNA expression in the aorta and 1.5-fold increases in the heart. Hypothyroid animals treated with T3 (5 microg/day for 6 days) or statin (0.2 mg/30 g for 2 weeks) reduce blood lipids and aortic OPN mRNA expression. Data obtained with ELISA analyses showed 1.5- and 1.7-fold increases in OPN protein in the aorta (10 weeks) and the heart (22 weeks), respectively. This increase is close to the mRNA expression in both tissues of hypothyroid mice. In addition, western blots showed several variants of OPN protein expressed in the aorta and the heart. The decrease in the 70 kDa OPN is accompanied by an increase in 45 kDa OPN in the aorta of hypothyroid mice. In contrast, only 45 kDa OPN is found in the heart of control and hypothyroid mice. These data indicate that the increase of OPN mRNA and protein expression occurs in cardiovascular tissues of hypothyroid mice. PMID- 16265596 TI - Glycoxidation of low-density lipoprotein promotes multiple apoptotic pathways and NFkappaB activation in human coronary cells. AB - Apoptosis of arterial cells induced by oxidized low-density lipoprotein (oxLDL) is thought to contribute to the progression of vascular dysfunction and atherogenesis. It is well established that diabetes mellitus is accompanied by both glycosylation and oxidation LDL, but the biological effects of these modified lipoproteins are poorly understood. We demonstrate here that glycosylated oxLDL (glc-oxLDL) promotes apoptotic signaling in human coronary smooth muscle cells. This was associated by a decrease of the antiapoptotic protein Bcl-2, an increase of the pro-apoptotic protein Bax, and activation of caspase 3. Glc-oxLDL also activated NFK: B and decreased IK: B, these effects were more pronounced than those achieved with oxLDL. Our study shows that glc oxLDL influences a broad cascade of signaling transduction pathways, which may not only result in apoptosis, but also could affect NFkappaB in human coronary cells. This cascade of events may influence the evolution of atherogenesis and vascular complications in diabetic patients. PMID- 16265598 TI - Bone remodeling at the iliac crest can predict the changes in remodeling dynamics, microdamage accumulation, and mechanical properties in the lumbar vertebrae of dogs. AB - We previously demonstrated that suppression of bone remodeling allows microdamage to accumulate, thereby leading to reduced bone toughness in dog bone. In this study we evaluated the relationships between bone remodeling at the iliac crest and skeletal activation frequency, microdamage accumulation, or biomechanical properties of lumbar vertebrae using the same dogs to determine whether bone remodeling at the iliac crest can predict damage accumulation and mechanical parameters of the lumbar spine following treatment with antiresorptive agents. Thirty-six female beagles, 1 to 2 years old, were divided into three groups. The control group was treated daily for 12 months with saline vehicle. The remaining two groups were treated daily with oral risedronate at a dose of 0.5 mg/kg/day, or alendronate at 1.0 mg/kg/day orally. The doses of these bisphosphonates were 5 to 6 times the clinical doses approved for treatment of osteoporosis in humans. After sacrifice, the right ilium and L2 vertebra were assigned to histomorphometry. The left ilium and L3 vertebra were used for microdamage analysis. The L4 vertebra was mechanically tested to failure in compression, and bone toughness calculated from the stress-strain curve. There was a strong positive relationship for activation frequency (Ac.f) between ilium and lumbar vertebrae (r2 = 0.82; P < 0.0001). Iliac crest Ac.f underestimates Ac.f in L2, but L2 Ac.f reaches a minimum threshold and does not decline further when iliac crest Ac.f is below 0.10/yr. Microdamage (Cr.S.Dn) accumulation at the ilium was significantly associated with increased microdamage accumulation in the L3 lumbar vertebra (r2 = 0.43, P < 0.0001). The data also show that bisphosphonate treatment increased Cr.S.Dn at a faster rate in L3 than in the iliac crest. Although bisphosphonate treatment decreased bone toughness in L4, this decrease demonstrated no relationship to decreased Ac.f in the ilium. These results clearly indicate that bone remodeling data obtained from iliac crest biopsy could be used to estimate the activation frequency and microdamage burden in the vertebral column. PMID- 16265597 TI - Translational neuroimaging: positron emission tomography studies of monoamine oxidase. AB - Positron emission tomography (PET) using radiotracers with high molecular specificity is an important scientific tool in studies of monoamine oxidase (MAO), an important enzyme in the regulation of the neurotransmitters dopamine, norepinephrine, and serotonin as well as the dietary amine, tyramine. MAO occurs in two different subtypes, MAO A and MAO B, which have different substrate and inhibitor specificity and which are different gene products. The highly variable subtype distribution with different species makes human studies of special value. MAO A and B can be imaged in the human brain and certain peripheral organs using PET and carbon-11 (half-life 20.4 minutes) labeled mechanism-based irreversible inhibitors, clorgyline and L -deprenyl, respectively. In this article we introduce MAO and describe the development of these radiotracers and their translation from preclinical studies to the investigation of variables affecting MAO in the human brain and peripheral organs. PMID- 16265599 TI - Epidemiology and risk factors analysis of elaphostrongylosis in red deer (Cervus elaphus) from Spain. AB - We studied the distribution and faecal shedding pattern of the first-stage larvae (L1) of Elaphostrongylus cervi (Nematoda: Protostrongylidae) in the red deer (Cervus elaphus) across Spain, where excretion was widespread. We evaluated the effects of individual, population and environmental factors on E. cervi L1 counts in 18 free-ranging red deer populations in South Central Spain. In this area, prevalence was 71.42+/-2.14% (n = 448) and mean intensity (n = 320) was 74.50+/ 10.35. Aggregation of deer at water-holes was positively associated with E. cervi L1 prevalence, possibly due to spatial and temporal odds of infected gastropods, red deer and infective E. cervi L1 larvae being encountered. Prevalence increased with age, and there was also a trend towards males having higher intensities than females. A slightly decreasing age-intensity profile was identified for females, which may suggest a role of acquired immunity. PMID- 16265600 TI - Effects of dietary leucine supplementation on exercise performance. AB - Branched chain amino acids (BCAA), particularly leucine, have been suggested to be ergogenic for both endurance and strength/power performance. This study investigated the effects of dietary leucine supplementation on the exercise performance of outrigger canoeists. Thirteen (ten female, three male) competitive outrigger canoeists [aged 31.6 (2.2) year, VO(2max) 47.1 (2.0) ml kg(-1) min(-1)] underwent testing before and after 6-week supplementation with either capsulated L: -leucine (45 mg kg(-1) d(-1); n = 6) or placebo (cornflour; n = 7). Testing included anthropometry, 10 s upper body power and work and a row to exhaustion at 70-75% maximal aerobic power where perceived exertion (RPE), heart rate (HR) and plasma BCAA and tryptophan concentrations were assessed. Leucine supplementation resulted in significant increases in plasma leucine and total BCAA concentrations. Upper body power and work significantly increased in both groups after supplementation but power was significantly greater after leucine supplementation compared to the placebo [6.7 (0.7) v. 6.0 (0.7) W kg(-1)]. Rowing time significantly increased [77.6 (6.3)-88.3 (7.3) min] and average RPE significantly decreased [14.5 (1.5)-12.9 (1.4)] with leucine supplementation while these variables were unchanged with the placebo. Leucine supplementation had no effect on the plasma tryptophan to BCAA ratio, HR or anthropometric variables. Six weeks' dietary leucine supplementation significantly improved endurance performance and upper body power in outrigger canoeists without significant change in the plasma ratio of tryptophan to BCAA. PMID- 16265601 TI - Ventilatory changes following head-up tilt and standing in healthy subjects. AB - Passive tilting increases ventilation in healthy subjects; however, controversy surrounds the proposed mechanism. This study is aimed to evaluate the possible mechanism for changes to ventilation following passive head-up tilt (HUT) and active standing by comparison of a range of ventilatory, metabolic and mechanical parameters. Ventilatory parameters (V (T), V (E), V (E)/VO(2), V (E)/VCO(2), f and PetCO(2)), functional residual capacity (FRC), respiratory mechanics with impulse oscillometry; oxygen consumption (VO(2)) and carbon dioxide production (VCO(2)) were measured in 20 healthy male subjects whilst supine, following HUT to 70 degrees and unsupported standing. Data were analysed using a linear mixed model. HUT to 70 degrees from supine increased minute ventilation (V (E)) (P<0.001), tidal volume (V (T)) (P=0.001), ventilatory equivalent for O(2) (V (E)/VO(2)) (P=0.020) and the ventilatory equivalent for CO(2) (V (E)/VCO(2)) (P<0.001) with no change in f (P=0.488). HUT also increased FRC (P<0.001) and respiratory system reactance (X5Hz) (P<0.001) with reduced respiratory system resistance (R5Hz) (P=0.004) and end-tidal carbon dioxide (PetCO(2)) (P<0.001) compared to supine. Standing increased V (E) (P<0.001), V (T) (P<0.001) and V (E)/VCO(2) (P=0.020) with no change in respiratory rate (f) (P=0.065), V (E)/VO(2) (P=0.543). Similar changes in FRC (P<0.001), R5Hz (P=0.013), X5Hz (P<0.001) and PetCO(2) (P<0.001) compared to HUT were found. In contrast to HUT, standing increased VO(2) (P=0.002) and VCO(2) (P=0.048). The greater increase in V (E) in standing compared to HUT appears to be related to increased VO(2) and VCO(2) associated with increased muscle activity in the unsupported standing position. This has implications for exercise prescription and rehabilitation of critically ill patients who have reduced cardiovascular and respiratory reserve. PMID- 16265602 TI - Tissue Doppler echocardiography in evaluation of cardiac effects of subclinical hypothyroidism. AB - OBJECTIVE: Subclinical hypothyroidism (SH) is a mild form of thyroid failure, associated with initial signs of cardiovascular hypothyroidism. Tissue Doppler echocardiography (TDE) is a new and powerful method in evaluation of both regional and global systolic or diastolic ventricular function. We aimed to investigate the use of TDE in evaluation of cardiac effects of SH and affect of thyroid hormone replacement therapy (TRT) on tissue Doppler parameters of SH patients. METHODS: Twenty-two patients who were diagnosed as SH and 22 healthy, age and sex-matched cases were included in the study. Conventional echocardiography and TDE were performed in all individuals. TRT was started in SH group. On the achievement of euthyroid state echocardiography were repeated. RESULTS: Septal annulus relaxation time was significantly higher in SH group (82+/-21, 98+/-11 ms, p=0.024). Lateral annulus and myocardial relaxation times, precontraction/contraction ratios and precontraction times were also slightly higher. Septal lateral annulus and lateral myocardial relaxation times were decreased after TRT (98+/-11 vs. 81+/-12, p<0.001, 89+/-14 vs. 78+/-11, p=0.022, 90+/-16 vs. 80+/-14 ms, p=0.008, respectively). Precontraction times and precontraction/contraction ratios decreased after TRT but did not reach the significance level. There was a positive correlation between TSH and TDE relaxation times. CONCLUSIONS: TDE is a powerful tool in diagnosis and follow-up of SH patients and TRT inhibits adverse affects of SH on myocardium. Septal myocardium is the most affected region of left ventricle in SH. The relaxation time is the best criteria of cardiac involvement and monitoring the effect of TRT. PMID- 16265603 TI - Influence of angiotensin II receptors blocking on overall left ventricle's performance of patients with acute myocardial infarction of limited extent. Echocardiographic assessment. AB - INTRODUCTION: Recent studies documented the beneficial effect of angiotensin receptor blockers (ARBs) on patients (pts) with acute myocardial infarction (AMI) combined with left ventricle (LV) systolic dysfunction. The present study intended to assess the impact of the ARB irbesartan, on the overall LV performance in pts with uncomplicated AMI of limited extent. METHODS: Forty consecutive pts with first inferior AMI (AMI-I) and preserved LV-systolic function were enrolled. They were allocated into two groups: (a) 20 pts received the conventional treatment of AMI-I and placebo (CT) and (b) 20 pts administered irbesartan additionally to the conventional treatment (IR). Twenty four healthy individuals of matching age and sex were recruited as control group (CG). Complete echocardiographic examination, Tei index of overall LV function and systolic blood pressure (SBP) were measured on the 8th post-infarct day. RESULTS: The Tei index of IR group (0.53+/-0.03) was significantly lower compared to that of CT group (0.78+/-0.05) (p<0.001) and was similar to that of CG (0.45+/ 0.03)(p=NS). Irbesartan induced a considerable decrease in both isovolumic relaxation (115+/-7 ms vs 140+/-7 ms; p<0.01) and contraction time (52+/-2 ms vs 64+/-3 ms; p<0.01) and a significant increase in ejection time (279+/-6 ms vs 256+/-8 ms; p<0.05). SBP in pts of IR group was similar to that of CT group (112+/-3 mmHg vs 113+/-4 mmHg; p=NS). CONCLUSIONS: Therapy with Irbesartan improves overall LV function of pts with AMI-I. Irbesartan leads to acceleration of the LV relaxation, which possibly indirectly ameliorates LV systolic performance too. This beneficial influence is possible attributed to a direct tissue effect of the drug and not to its hemodynamic action. PMID- 16265604 TI - An unexpected cause of angina detected by ECG-gated cardiac computed tomography. AB - Until recently anomalous coronary artery anatomy was only identified either by coronary angiography, at autopsy, or during cardiac surgery. With recent developments in the area of cardiac imaging, ECG-gated cardiac computed tomography (CT) has emerged as a minimally invasive modality to delineate both coronary anatomy and pathology. We present a case of an anomalous right coronary artery origin from the ascending aorta detected by ECG-gated cardiac CT in a 47 year-old male who presented to the emergency department complaining of acute chest pain after intense exercise. Given its relatively non invasive nature, ECG gated cardiac CT may assist in the diagnosis and management of patients with atypical chest pain in which more invasive diagnostic examinations (i.e. coronary angiography) are not warranted. PMID- 16265605 TI - Medical management of HIV-hepatitis C virus coinfection in injection drug users. AB - Several million people inject drugs of abuse and, as a result, are coinfected with human immunodeficiency virus (HIV) and hepatitis C virus (HCV). The treatment of this coinfected drug-abusing population is fraught with many problems such that clinicians and other health care providers have to determine whether patients should be treated first for drug addiction, for HIV/AIDS, or for HCV infection or simultaneously treated. These proceedings present the incidence and prevalence of coinfections with HIV and HCV in high-risk populations and discuss the underlying pathophysiology of coinfections and the problems and strategies of managing the treatment of coinfections among people who also inject illicit drugs. In addition, the expert panel recommended further research to determine the best possible treatment regimens applicable to injection drug users coinfected with HIV and HCV. PMID- 16265606 TI - New drug targets for HIV and hepatitis C virus coinfection. AB - Current interferon (IFN)-based therapies for hepatitis C in patients coinfected with human immunodeficiency virus (HIV) and hepatitis C virus (HCV) may be limited by incomplete virological response, lack of adherence, and poor tolerability. Newer therapies for hepatitis C will target viral replication (e.g., HCV serine protease inhibitors, helicase inhibitors, RNA interference, or an HCV polymerase inhibitor). Other treatments will focus on viral translation (e.g., antisense molecules). Additions to IFN therapy that can modulate the immune response (e.g., thymosin, isatorbine, or injectable histamine) may improve tolerability of treatment. There need to be targets that minimize the inflammatory response by the liver (e.g., IFN-gamma). There are some therapeutic vaccines in early development. Drugs to replace or enhance ribavirin are being studied with IFN-based treatments. Strategic treatment trials that address sequencing of HCV and HIV therapy with current and future therapeutic agents and combination therapy need to be undertaken. PMID- 16265607 TI - Effect of alcohol use and highly active antiretroviral therapy on plasma levels of hepatitis C virus (HCV) in patients coinfected with HIV and HCV. AB - BACKGROUND: The interactions between human immunodeficiency virus (HIV), hepatitis C virus (HCV), alcohol, and antiretroviral therapy are complex. METHODS: We retrospectively assessed persons coinfected with HIV and HCV who achieved HIV suppression to < 500 copies/mL and continued to take antiretrovirals for > or = 6 months. Frozen plasma specimens were retrieved for quantitation of HCV RNA levels at baseline and 3, 6, and 12 months after beginning antiretroviral treatment. RESULTS: Median HCV RNA levels increased (0.35 log10 IU/mL) at month 3 (n = 44). HCV RNA levels decreased to below baseline by 12 months in patients consuming < 50 g of alcohol/day, whereas patients consuming > or = 50 g/day had a sustained increase (> 0.6 log10 IU/mL) from baseline (P = .04). CONCLUSIONS: Because low levels of HCV RNA are predictive of a virological response to therapy for HCV infection, it may be advantageous to first achieve suppression of HIV RNA and then initiate treatment for HCV infection in patients coinfected with HIV and HCV. Excess alcohol consumption with therapy for HIV infection increases HCV RNA levels and may impede the effectiveness of this treatment strategy. PMID- 16265608 TI - Acute hepatitis C virus infection. AB - Acute infection with hepatitis C virus (HCV) is a frequent but underrecognized problem among substance users, because it produces few symptoms. Despite this fact, a good deal has been learned recently from studies of cohorts of persons with acute HCV infection. Intensive study of these cohorts has suggested that there is a higher frequency of spontaneous clearance among persons with symptomatic infection and persons with vigorous adaptive immune responses. Similarly, polymorphisms for genes involved in innate immunity also appear to influence the outcome of acute HCV infection. Data on injection drug users with repeated exposures to HCV reveal the presence of partially protective immunity, which suggests that vaccine-based approaches may be feasible. Finally, antiviral therapy with interferon-based regimens for acute HCV infection produces significantly higher sustained virological responses than observed for chronic infection. Further work is needed to develop more-accurate assays for acute HCV infection, to define host and viral factors that predict outcome and to define the optimal duration and regimen of therapy. PMID- 16265610 TI - Cellular immune responses against hepatitis C virus. AB - Cellular immune responses are typically important in recovery from acute infections, and studies of acute hepatitis C confirm that broadly directed CD4+ and CD8+ T cell responses are associated with spontaneous clearance of infection. However, a major unanswered question is what role the cellular immune response plays in progression of liver disease during chronic infection. Classic models of hepatitis C suggest that cellular immune responses promote liver injury, either by causing direct cytolysis of infected cells or by promoting inflammation. However, clinical evidence suggests that persons with cellular immune dysfunction, such as that due to with human immunodeficiency virus (HIV) infection, have more-rapid disease progression. Recent data suggest that cellular immune responses do serve to limit the progression of liver disease, even if they are ineffective at clearance of virus. There is limited information on the effect of HIV coinfection on the cellular immune response to hepatitis C virus, but further study of this issue might shed light on the pathogenesis of liver disease in both immunocompromised and nonimmunocompromised hosts. PMID- 16265609 TI - Coinfection with HIV and hepatitis C virus among injection drug users in southern China. AB - BACKGROUND: We sought to examine coinfection with human immunodeficiency virus (HIV) and hepatitis C virus (HCV) among injection drug users (IDUs) in Guangxi, China. METHODS: A longitudinal cohort of IDUs (547 subjects) was established to study risk factors for bloodborne infections. At each visit, participants completed questionnaires defining demographic characteristics, patterns of drug use, and sexual behaviors. Blood samples were collected and analyzed for the presence and genotype of HIV and HCV. RESULTS: Coinfection with HIV and HCV was found in 17.6% of the IDUs. HCV was present in 95.1% of HIV-positive and 70.4% of HIV-negative heroin users. The prevalence of HIV in HCV-positive and HCV-negative heroin users was 23.4% and 3.6%, respectively. Multivariate logistic regression analysis revealed that sexual activity during the past 6 months and duration of injection drug use were significantly associated with coinfection with HIV and HCV. The main circulating HCV genotypes included 6a (38%), 3b (37%), and 1a (19%), whereas genotypes 6e (4%), 3a (2%), and 1b (1%) were present in only a few IDUs. Multiple HCV genotypes were present at each study site and did not segregate by HIV status or subtype. CONCLUSIONS: HCV is highly prevalent in IDUs throughout southern China. In Guangxi, HIV infections are the result of parenteral and sexual transmission, and, therefore, all IDUs are at high risk of coinfection with HIV and HCV. Molecular tracking of HCV may be a more sensitive predictor of the future spread of the HIV-1 epidemic than is HIV subtyping. This study emphasizes that, without implementation of injection prevention and primary substance abuse programs in China, the extent and effect of coinfection with HIV and HCV will only increase. PMID- 16265611 TI - Molecular mechanism of hepatic injury in coinfection with hepatitis C virus and HIV. AB - We have previously shown that hepatocytes exposed to hepatitis C virus (HCV) and human immunodeficiency virus (HIV) envelope proteins undergo apoptosis. In this article, we further elucidate the signaling mechanisms that mediate this effect. We found that, in human hepatocellular carcinoma (HepG2) cells, HCV E2 protein and HIV glycoprotein (gp) 120 significantly up-regulated the Fas ligand (FasL) and enhanced the formation of the Fas death-inducing signaling complex downstream of Fas receptor activation. Moreover, after stimulation with HCV E2 and HIV gp120, enhanced expression of caspases 2 and 7 and increased caspase 3 activity were observed. In addition, we showed up-regulation of the proapoptotic molecule Bid and its association with caspase 8 after treatment with these envelope proteins. We also found that HCV E2 and HIV gp120 induced a partial translocation of Bid to the mitochondria, which resulted in the release of cytochrome C and the apoptosis-inducing factor. Thus, the results of this study suggest that FasL and Bid play an important role in HCV and HIV envelope protein-induced apoptosis. PMID- 16265613 TI - Investigating neurocognitive features of hepatitis C virus infection in drug users: potential challenges and lessons learned from the HIV literature. AB - Studies of neurocognition in drug users with hepatitis C virus (HCV) infection are urgently needed but face many challenges. We review similarities between human immunodeficiency virus (HIV) and HCV infection in their neurocognitive features, discuss challenges associated with research among drug users, advocate that the HIV literature can usefully inform studies of HCV, and review findings by our group on neurocognition among substance users with HIV and/or HCV infection. PMID- 16265612 TI - Neuropsychological aspects of coinfection with HIV and hepatitis C virus. AB - Infection with hepatitis C virus (HCV) is commonly seen in persons with human immunodeficiency virus (HIV) infection, because the viruses share risk factors for transmission; coinfection is a leading cause of morbidity and mortality among HIV-infected persons. Neuropsychological consequences of HIV infection are well established, and studies of HCV-infected persons have revealed neuropsychiatric dysfunction in this population as well. Investigators now are focusing on neuropsychological sequelae of coinfection with HIV and HCV, and preliminary results suggest that coinfection has a possible deleterious effect on global cognitive functioning consistent with frontal-subcortical dysfunction. Data on neuropsychiatric symptoms in coinfected persons are inconclusive at this time and are complicated by important differences in study populations (e.g., injection drug use and disease severity). This review summarizes what is known about neuropsychological aspects of monoinfection with HIV and HCV, as well as coinfection, discusses implications of these findings, and suggests future directions for this research area. PMID- 16265614 TI - Outcomes among patients with end-stage liver disease who are coinfected with HIV and hepatitis C virus. AB - In at-risk populations, shared routes of transmission lead to high rates of concordance between infection with human immunodeficiency virus (HIV) type 1 and hepatitis C virus (HCV). In the era of highly active antiretroviral therapy (HAART), end-stage liver disease (ESLD) has emerged as a leading cause of mortality in coinfected patients. HAART-related toxicities have been implicated, especially when given to patients with viral hepatitis. Rates of response to treatment for HCV infection in coinfected patients continue to lag behind those in monoinfected patients, even with the advent of pegylated interferons. Liver transplantation has been approached with caution in this population because of concern about the sequelae of immunosuppression and HAART-related hepatotoxicity, and results have been conflicting. Clinical and serological markers of ESLD in coinfected patients, management of cirrhosis, and the appropriateness of transplantation are discussed. PMID- 16265615 TI - Delivering therapy for hepatitis C virus infection to incarcerated HIV seropositive patients. AB - The increase in morbidity and mortality due to end-stage liver disease has fueled recent guidelines that recommend consideration of treatment for hepatitis C in human immunodeficiency virus (HIV)-infected patients. Unfortunately, studies indicate that few patients coinfected with HIV and hepatitis C virus (HCV) are treated for their underlying hepatitis because of ongoing substance abuse, depression, chaotic lifestyles, homelessness, and perceived nonadherence. The structured environment of the prison system enables clinicians to provide complicated therapies for HCV to HIV-infected patients in combination with substance abuse programs. Furthermore, adherence to and adverse effects of therapy can be closely monitored. Offering treatment for HCV infection during incarceration to HIV-seropositive persons is highly efficient and targets underserved minority patients who have limited access to care in the community. PMID- 16265616 TI - Clinical trials of treatment for hepatitis C virus infection in HIV-infected patients: past, present, and future. AB - Coinfection with hepatitis C virus (HCV) and human immunodeficiency virus (HIV) type 1 is common. Rates of liver-related morbidity and mortality have been increasing in the coinfected population, and treatment for HCV infection in this group remains a challenge. The HCV-monoinfected population, especially patients infected with HCV genotype 2 or 3, has benefited dramatically from the advent of treatment with pegylated interferon plus ribavirin; rates of sustained virological response approach 55%. Coinfected patients lag behind, with rates of sustained virological response ranging between 26% and 40%; rates of sustained virological response are even lower among patients infected with HCV genotype 1. It is encouraging, however, that therapies known to be safe for treating monoinfected patients have been proven to be generally safe and well tolerated in patients coinfected with HIV and HCV, as well. Future therapies, some of which are currently in development, will likely include new targets, such as helicase and polymerase. It is hoped that, as more-effective agents are discovered, the disparity in treatment response will diminish. PMID- 16265617 TI - Therapeutic interventions for HIV infection and chronic viral hepatitis. AB - Combination antiretroviral therapy reduces overall and liver-specific morbidity and mortality in coinfection with human immunodeficiency virus (HIV) and hepatitis C virus (HCV) and represents the most beneficial pharmaceutical treatment intervention for most coinfected patients. Antiviral therapy for HCV infection is potentially organ- and life-saving but, in general, should be reserved for patients who achieve suppression of HIV RNA and immune restoration from combination antiretroviral therapy or for patients with nadir CD4+ T lymphocyte levels of >350 cells/microL. Safe and virologically active treatment of coinfection with HIV and hepatitis B virus can be concurrently achieved by the use of combination antiretroviral therapy regimens containing lamivudine and/or tenofovir. PMID- 16265618 TI - Coinfection with HIV and hepatitis C virus in injection drug users and minority populations. AB - Coinfection with human immunodeficiency virus (HIV) and hepatitis C virus (HCV) is common. In the United States, it has been estimated that 25% of persons infected with HIV are also infected with HCV. The prevalence of coinfection with HIV and HCV is highest among those infected via percutaneous routes. In fact, in urban areas in the United States, 50%-90% of persons infected with HIV via injection drug use are coinfected with HCV. In addition, limited data from drug treatment centers in these urban areas suggest that the prevalence of coinfection with HIV and HCV may be highest among African Americans and Hispanics. Little information is available with regard to the epidemiology of coinfection with HIV and HCV among injection drug users (IDUs) or minority populations. Likewise, although there is a growing body of data on the potential complexities of treating HCV among IDUs and the poor response to current anti-HCV treatment among African Americans, few data address the therapy of coinfection with HIV and HCV among IDUs and minority populations. PMID- 16265619 TI - A systematic approach to the treatment of HIV and hepatitis C virus infection in the inner city: a Canadian perspective. AB - The treatment of injection drug users (IDUs) coinfected with human immunodeficiency virus (HIV) and hepatitis C virus (HCV) presents multiple challenges, many of which could be addressed by the development of directly observed therapy programs. This is made all the more feasible by the validation of once-daily treatment regimens for HIV. We have demonstrated that virological suppression can be achieved and maintained in as many as 80% of active IDUs who have received highly active antiretroviral therapy for 48 months. This approach has now been validated in first- and second-line therapy, as well as for the treatment of bacterial infections in this population, achieving therapeutic results similar to those reported in the general population. The model is now being applied to the treatment of HCV infection, focusing on patients with infection due to HCV genotype 2 or 3, in whom the likelihood of response may exceed 80%. Our ultimate goal is to ensure that, even in treating IDUs in the inner city, no patient is left behind. PMID- 16265620 TI - Approaching treatment for hepatitis C virus infection in substance users. AB - Although injection drug users represent the majority of incident and prevalent cases of hepatitis C, most lack access to treatment because of concerns about adherence, treatment efficacy, and reinfection. On the basis of an increasing body of evidence suggesting that injection drug users can successfully undergo treatment for hepatitis C virus (HCV) infection, the 2002 National Institutes of Health Consensus Statement on Hepatitis C has recommended that substance users, even those with ongoing drug use, be considered for treatment for HCV infection on a case-by-case basis. However, the criteria on which these treatment decisions should be based are unclear: The duration of pretreatment drug abstinence, comorbid psychiatric illness, intercurrent drug use, and the potential for injected interferon to cause relapse of drug use may all influence results of treatment for HCV infection. This overview summarizes my group's current data about treatment for HCV infection in substance users and the effect of these potential barriers on outcomes of treatment. PMID- 16265621 TI - Integrating care for hepatitis C virus (HCV) and primary care for HIV for injection drug users coinfected with HIV and HCV. AB - Injection drug use accounts for most of the incident infections with hepatitis C virus (HCV) and for at least one-third of new human immunodeficiency virus (HIV) infections. Coinfection with HCV and HIV presents complex and challenging medical conditions. Ensuring access to and maintaining care for HIV and HCV for drug users presents special challenges to the health care team that require a nonjudgmental attitude, experience, and patience. Care for HCV infection, however, can be used as an instrument to engage drug-using persons in ongoing primary care relationships. Common elements to both care for HCV infection and primary care for HIV infection are testing for and counseling about HCV and HIV, substance abuse and mental health services, social support, and subspecialty referral. These elements, in particular treatment for substance abuse, can be focal points for model care systems that provide integrative care for both HCV and HIV infections. PMID- 16265622 TI - Treatment of opioid dependence and coinfection with HIV and hepatitis C virus in opioid-dependent patients: the importance of drug interactions between opioids and antiretroviral agents. AB - The occurrence of human immunodeficiency virus (HIV) disease and hepatitis C is common in injection drug users, most of whom are opioid dependent. Methadone pharmacotherapy has been the most widely used treatment for opioid addiction in this population. Methadone has significant, adverse drug-drug interactions with many antiretroviral therapeutic agents that can contribute to nonadherence and poor clinical outcomes in this high-risk population. The present article summarizes current knowledge about interactions between methadone and antiretroviral medications. Buprenorphine is the newest agent available for the treatment of opioid dependence and may have fewer adverse interactions with antiretroviral agents. Buprenorphine has a significant pharmacokinetic interaction with efavirenz but no pharmacodynamic interaction; therefore, simultaneous administration of these drugs is not associated with opioid withdrawal, as has been observed with methadone. This promising finding may simplify the treatment of opioid-dependent patients with HIV disease and should also improve clinical outcomes for persons coinfected with HIV and hepatitis C virus. PMID- 16265623 TI - New drug targets for HIV. AB - A significant number of human immunodeficiency virus (HIV) infections have become resistant to antiretroviral treatment, which means that there is a paramount need for novel drug targets to defeat the virus. Until recently, all HIV drugs inhibited HIV replication by mechanisms operating inside infected cells. In contrast, new antiretroviral drugs operate outside infected cells. Their mechanism of action consists in inhibiting entry of the virus into cells, thereby halting the very first step of HIV replication. Examples of this new class of drugs include entry inhibitors, coreceptor antagonists, and fusion inhibitors. In addition to their novel mechanism of action, this new class of drugs also has potential action against drug-resistant HIV strains, causes minimal adverse effects, and may be administered in a simplified, once-daily dosing regimen. New classes of anti-HIV drugs--and new drugs in existing classes--represent the best hope for people infected with HIV, especially those who have exhausted current therapies. PMID- 16265624 TI - Classics in anatomy: Keith Porter's Harvey Lecture, "the submicroscopic morphology of protoplasm". PMID- 16265625 TI - Tour of organelles through the electron microscope: a reprinting of Keith R. Porter's classic Harvey Lecture with a new introduction. PMID- 16265626 TI - Induction of Bcl-2 and Bax was related to hyperphosphorylation of tau and neuronal death induced by okadaic acid in rat brain. AB - Abnormal hyperphosphorylation of the cytoskeletal protein tau is a characteristic feature of neurodegeneration in Alzheimer's disease (AD) brain. Okadaic acid (OA), a protein phosphatase inhibitor, induces neuronal death and hyperphosphorylation of tau. In the present study using a model of microinjection of OA into rat frontal cortex, we aimed to investigate if OA-induced hyperphosphorylation of tau and neuronal death are related to the expression of Bcl-2, an apoptosis inhibitor, or Bax, an apoptosis inducer. Immunohistochemistry and Western blot analysis showed that OA injection dose- and time-dependently induced the expression of Bcl-2 and Bax protein in the surrounding of OA injection areas, which were similar with that of AT8 immunostaining, a marker of hyperphosphorylated tau. However, the ratios of Bcl-2 over Bax had a negative relationship to the expression of AT8. Furthermore, double fluorescent staining showed that AT8-positive neurons mainly costained with terminal deoxynucleotidyl transferase-mediated deoxyuridinetriphosphate nick-end labeling, a marker of DNA damage, indicating that tau hyperphosphorylation may be associated with DNA damage in the neurons of rat brain. In the areas more adjacent to the OA injection site, most neurons with AT8-positive staining showed vulnerability to OA toxicity and could be triple-stained with Bcl-2 and Bax or double-stained with Bcl-2. However, in the areas further from the OA injection site, neurons with few AT8-positive staining showed resistance to OA toxicity and only stained with Bcl 2, but not Bax. The results suggest that the ratios of Bcl-2 over Bax expression may have an effect on tau hyperphosphorylation and neuronal death following OA injection. PMID- 16265627 TI - Multicomponent variance estimation for binary traits in family-based studies. AB - In biometrical genetic analyses of binary traits, the use of family data overcomes some limitations of twin studies, particularly in terms of sample size and types of genetic or environmental factors that can be estimated. However, because of computational problems, recent methods in the application of generalized linear mixed models for family data structure have limited the ability to handle large data sets with general covariates. In this paper, we investigate the use of the hierarchical likelihood approach to the analysis of binary traits from family data. In a simulation study, the method is shown to be highly accurate for the estimation of both the variance components and fixed regression parameters, even for small family sizes. For illustration, we analyze a real data set of familial aggregation of preeclampsia, a pregnancy-induced hypertension. When possible, the analysis is compared with the exact maximum likelihood approach. PMID- 16265628 TI - Plasmapheresis for diffuse alveolar hemorrhage in a patient with Wegener's granulomatosis: case report and review of the literature. AB - We present a case of a 20-year-old male with Wegener's Granulomatosis involving the upper respiratory tract, lungs, and kidneys. In his fourth hospital admission, the patient presented with diffuse alveolar hemorrhage and poor pulmonary function: FiO2 of 100% and PEEP of 17cm H2O on intubation. Due to a fast clinical deterioration while receiving drug therapy (cyclophosphamide and methylprednisolone), we performed nine daily 1-volume therapeutic plasma exchanges (TPE) using 5% albumin as replacement fluid. TPE resulted in a decrease in cytoplasmic anti-neutrophil cytoplasm antibodies (c-ANCA) titer from 1:1,024 to 1:16. On the ninth day of plasmapheresis, his pulmonary status was markedly improved with FiO2 of 60% and PEEP of 8 cm H2O. The patient was later extubated and discharged home in stable condition. Wegener's Granulomatosis with pulmonary hemorrhage is not included in the current guidelines for therapeutic apheresis; therefore, we report this case and, if warranted, propose this condition to be included in the guidelines. PMID- 16265629 TI - Increased production of soluble HLA-G molecules in stimulated peripheral blood mononuclear cells following extracorporeal photopheresis: is it a mechanism involved in the therapeutic effect of the procedure? AB - We hypothesized that the effects of extracorporeal photopheresis (ECP) are mediated by induction of immunosuppressive cytokines like IL-10, which enhances synthesis of HLA-G molecules. HLA-G products are expressed by CD14+ peripheral blood mononuclear cells (PBMC) and play an important role in inhibition of cell mediated immunity. ECP induces apoptosis in lymphocytes but not in CD14+ cells. We, therefore, investigated the concentrations both of IL-10 and of soluble HLA G5/sHLA-G1 molecules in supernatants from cultures of lipopolysaccharide stimulated PBMC obtained from leukocyte collection bags of 10 patients receiving ECP for graft versus host disease both before (pre-irradiation) and after (post irradiation) exposure to 8-methoxypsoralen and UVA irradiation. Levels of both IL 10 and HLA-G5/sHLA-G1 molecules were increased in the post-irradiation cultures. This suggests that therapeutic effects of ECP could be mediated by increased production of IL-10 and tolerogenic HLA-G molecules. PMID- 16265630 TI - Acute hemolytic transfusion reaction in a pediatric patient following transfusion of apheresis platelets. AB - The practice of transfusing ABO-incompatible platelets, driven primarily by concerns about inventory management, has been considered generally safe because the accompanying plasma is usually diluted in the recipient's total blood volume. However, if the platelet product contains a large volume of plasma or a high concentration of incompatible isoagglutinin, there may be hemolysis of the recipient's red cells. Patients with a small blood volume, such as babies and children, are considered to be at particular risk for such a complication. We describe the case of a baby who suffered massive hemolysis of her group A red cells after transfusion of group O Apheresis Platelets containing a high-titered anti-A isoagglutinin. We also offer a review of the literature on this subject and recommendations to avoid acute hemolytic reactions as a result of platelet transfusion. PMID- 16265631 TI - Role of calcium-dependent actin-bundling proteins: characterization of Dictyostelium mutants lacking fimbrin and the 34-kilodalton protein. AB - Actin-bundling proteins organize actin filaments into densely packed bundles. In Dictyostelium discoideum two abundant proteins display calcium-regulated bundling activity, fimbrin and the 34-kDa protein (ABP34). Using a GFP fusion we observed transient localization of fimbrin at the phagocytic cup and macropinosomes. The distribution of truncated constructs encompassing the EF hands and the first actin-binding domain (EA1) or both actin-binding domains devoid of EF hands (A1A2) was indistinguishable from that of the full length protein. The role of fimbrin and a possible functional overlap with ABP34 was investigated in fim- and double 34-/fim- mutants. Except for a moderate cell size defect, fim- mutants did not show defects in growth, endocytosis, exocytosis, and chemotaxis. Double mutants were characterized by a small cell size and a defect in morphogenesis resulting in small fruiting bodies and a low spore yield. The cell size defect could not be overcome by expression of fimbrin fragments EA1 or A1A2, suggesting that both bundling activity and regulation by calcium are important. Induction of filopod formation in 34-/fim- cells was not impaired, indicating that both proteins are dispensable for this process. We searched in the Dictyostelium genome database for fimbrin-like proteins that could compensate for the fimbrin defect and identified three unconventional fimbrins and two more proteins with actin-binding domains of the type present in fimbrins. PMID- 16265632 TI - Characterization of proteoglycan depletion in articular cartilage using two dimensional time domain nuclear magnetic resonance. AB - In vitro proteoglycan (PG) depletion in the 20-40% range (enzymatic PG depletion of normal cartilage in the early osteoarthritis (OA) PG depletion range) was investigated in articular cartilage using 2D time domain NMR relaxation techniques. Spin-lattice relaxation times were measured at low fields (T(1rho)) and at high fields (T(1)) using nonselective and selective excitation pulse sequences. The short relaxation time magnetization components in T(1rho) ( approximately 8% signal) and nonselective T(1) ( approximately 5% signal) experiments were significantly altered with PG degradation. In addition, a magnetization component ( approximately 5% signal) with a "fast " T(1) approximately 7 ms was observed in the T(1) experiment involving selective excitation. This fast T(1) was at least 10 times shorter than the short T(1) in the nonselective experiment and was associated with a strong magnetization exchange mechanism between collagen and PG. The results suggest that T(1rho) and T(1) (nonselective and selective) relaxation based MRI techniques, which focus on the short relaxation time magnetization components, have the potential of detecting molecular abnormalities associated with early OA earlier than single, long relaxation time component approaches. PMID- 16265633 TI - Assessment of glutamate and glutamine contribution to in vivo N-acetylaspartate quantification in human brain by (1)H-magnetic resonance spectroscopy. AB - N-Acetylaspartate (NAA) is one of the most important metabolites detectable by brain (1)H-MRS being considered an index of neuronal integrity. At the low magnetic field used in most clinical settings beta,gamma-glutamate/glutamine (Glx) resonances are very close and partially overlap the methyl-NAA resonance interfering with NAA quantification especially at low TE and in the presence of increased Glx signals. NAA overestimation due to Glx on a set of model solutions containing NAA, glutamate, and glutamine in variable amounts was evaluated and the result tested in vivo in six healthy controls and five age- and sex-matched patients with hepatic encephalopathy (HE), the latter having an increased Glx content. A method to assess in vivo the NAA overestimation caused by Glx is proposed. A perfect match was obtained between the assessment of Glx contamination on the NAA of healthy controls and that obtained on the model solutions. However, a substantial difference in NAA overestimation was found between controls and HE patients that cannot be explained by our model. An interpretative hypothesis is provided. PMID- 16265634 TI - T(2) + measurement during acute cerebral ischemia by Carr-Purcell MRI at 4T. AB - Metabolic and structural changes occur in brain tissue within minutes of ischemia. The adiabatic multi-echo (Carr-Purcell) localization pulse sequence LASER has shown promise in detecting tissue contrast changes within the first hour of ischemia. The purpose of this initial study was to combine the LASER localization sequence with fast 3D echo-planar imaging (EPI) to quantify the regional apparent transverse relaxation (T(2) (dagger)) in a rabbit model of acute embolic ischemia at 4 Tesla. Carr-Purcell T(2) (dagger)-weighted images were acquired at 7 different echo-times and used to estimate T(2) (dagger) in both cortex and striatum. In ischemic tissue identified by 2,3,5 triphenyltetrazolium chloride (TTC) staining, the T(2) (dagger) increased by approximately 31% after 1 hour of ischemia and remained elevated until study completion at 4 h of ischemia. Lesion volume, defined as the number of pixels with T(2) (dagger) greater than 90 ms, increased by 40% between 1 and 4 h after induction of ischemia. Carr-Purcell LASER-EPI T(2) (dagger)-weighted images show promise in detecting early tissue changes in focal cerebral ischemia. PMID- 16265635 TI - Single heartbeat cardiac tagging for the evaluation of transient phenomena. AB - Many cardiac abnormalities are of a transient nature, creating a beat-to-beat variation in myocardial function. This work presents the cardiac imaging technique for the measurement of regional function during transient cardiac phenomena. All information necessary for the reconstruction of a cine loop is acquired within a single heartbeat, avoiding the temporal blurring introduced by segmented imaging due to the assumption of cardiac cycle periodicity. This method incorporates a gradient-optimized, high-efficiency EPI-SSFP sequence and TSENSE parallel imaging. For acquisitions with readout resolutions of 128,160, 192, and 256 points, the technique produced images with average temporal resolution of 35, 39, 43, and 52 ms and average spatial resolutions of 2.65, 2.12, 1.77, and 1.32 mm in the readout direction, respectively, and 2.88 and 2.08 mm in the phase encode direction for acceleration rates of 3 and 4, respectively. Local apparent strains in the single slice and measurements of ventricular end-systolic and end diastolic areas were used as quantitative measures to validate the single heartbeat technique. To demonstrate the utility of the sequence, movie loops were acquired for multiple heartbeats in non-breath-held acquisitions as well as during a Valsalva maneuver. A heartbeat-interleaved acquisition allowed for the reconstruction of nonaccelerated images from R contiguous heartbeats. Images reconstructed from such data displayed tag blurring and reduced tag persistence due to motion and inter-heartbeat variability. Images acquired during the Valsalva maneuver demonstrated apparent beat-to-beat variability, visible both in the images and as changing strain patterns and ventricular volumes. PMID- 16265636 TI - A novel bioactive porous CaSiO3 scaffold for bone tissue engineering. AB - The aim of this study was to fabricate bioactive porous CaSiO3 scaffolds and examine their effects on proliferation and differentiation of osteoblast-like cells. In this study, porous CaSiO3 scaffolds were obtained by sintering a ceramic slip-coated polymer foam at 1350 degrees C. X-ray diffraction (XRD) of the scaffolds indicated that the products were essentially pure alpha-CaSiO3. The obtained scaffolds had a well-interconnected porous structure with pore sizes ranging from several micrometers to more than 100 microm and porosities of 88.5 +/- 2.8%. The in vitro bioactivity of the scaffolds was investigated by soaking them in simulated body fluid (SBF) for 7 days and then characterizing them by scanning electron microscopy (SEM) and energy-dispersive spectroscopy (EDS) analysis. The results indicated that hydroxyapatite (HAp) was formed on the surface of the scaffolds. In addition, the scaffolds were incubated in Ringer's solution at 37 degrees C to study the in vitro degradation by measurement of weight loss after incubation, which showed that the CaSiO3 scaffolds were degradable. The cellular responses to the scaffolds were assessed in terms of cell proliferation and differentiation. Osteoblast-like cells were seeded into the CaSiO3 scaffolds. SEM observations showed that there was significant cell adhesion, as the cells spread and grew in the scaffolds. In addition, the proliferation rate and alkaline phosphatase (ALP) activity of the cells in the scaffolds were improved as compared to the controls. These studies demonstrate initial in vitro cell compatibility and their potential application to bone tissue engineering. PMID- 16265637 TI - Synthesis, characterization, and osteocompatibility evaluation of novel alanine based polyphosphazenes. AB - This study deals with the synthesis and in vitro osteocompatibility evaluation of two novel alanine-containing biodegradable ester polyphosphazenes as candidates to form self-setting composites with hydroxyapatite (HAp) precursors. The two novel biodegradable polyphosphazenes synthesized were poly[(ethyl alanato)1.0(ethyl oxybenzoate)1.0 phosphazene] (PN-EA/EOB) and poly[(ethyl alanato)1.0(propyl oxybenzoate)1.0 phosphazene] (PN-EA/POB). The polymers were characterized by multinuclear magnetic resonance (NMR), differential scanning calorimetry (DSC), and gel permeation chromatography (GPC). Biodegradability and percentage water absorption of the polymers were evaluated by following the mass change in phosphate buffer (pH 7.4) at 37 degrees C. PN-EA/POB underwent faster degradation and showed higher water absorption compared to PN-EA/EOB. Both polymers became insoluble in common organic solvents following hydrolysis presumably due to crosslinking reactions accompanying the degradation process. Osteoblast cell adhesion and proliferation on PN-EA/EOB and PN-EA/POB was followed by scanning electron microscopy (SEM) and by using a biochemical assay. Both PN-EA/EOB and PN-EA/POB supported the adhesion and proliferation of primary rat osteoblast cells in vitro. Furthermore, the enzymatic activity of the osteoblast cells cultured on the polymers was confirmed by the alkaline phosphatase activity. Thus, these biodegradable amino-acid-based polyphosphazenes are promising new materials for forming self-setting bone cements. PMID- 16265638 TI - Enhancement of in vivo endothelialization of tissue-engineered vascular grafts by granulocyte colony-stimulating factor. AB - Successful reconstruction of large-diameter blood vessel in humans has been demonstrated using the tissue engineering technique, but improvement in patency of small-diameter bioartificial vascular graft remains a great challenge. This study reports that granulocyte colony-stimulating factor (G-CSF) can enhance in vivo endothelialization of tissue-engineered vascular grafts, which could be used to improve patency of small-diameter vascular graft. Vascular grafts were tissue engineered with decellularized canine abdominal aortas and canine autologous bone marrow-derived cells. Prior to cell seeding onto decellularized graft matrices, bone marrow-derived cells were induced to differentiate into endothelial cells and smooth muscle cells. The cell-seeded vascular grafts were implanted into the abdominal aortas of bone marrow donor dogs. Before and after graft implantation, G-CSF was administered subcutaneously to the dogs (n = 3). The grafts implanted into the dogs not receiving G-CSF were used as controls (n = 3). Eight weeks after implantation, grafts in both groups showed regeneration of vascular tissues including endothelium and smooth muscle. Importantly, endothelium formation was more extensive in the G-CSF-treated grafts than in the control grafts, as assessed with reverse transcription polymerase chain reaction, western blot, and immunohistochemistry. In addition, intimal hyperplasia was significantly reduced in the G-CSF-treated grafts compared to the control grafts. This study suggests that G-CSF administration could be applied to improve patency of small-diameter tissue-engineered vascular grafts. PMID- 16265639 TI - Prenatal exposure to fluconazole: an identifiable dysmorphic phenotype. AB - BACKGROUND: Fluconazole is a triazole antifungal used to treat mycotic infections. Fluconazole is reported to act as a teratogen when used continuously at a dosage of 400-800 mg daily. Fluconazole embryopathy was previously reported in 4 cases. The common features that were also seen in the current case include multiple synostosis (including craniosynostosis and digital synostosis), congenital heart defects, skeletal anomalies, and recognizable dysmorphic facial features. CASE: We report the case of a 9-month-old male born to a 30-year-old woman following a 37-week pregnancy. The pregnancy was complicated by maternal human immunodeficiency virus (HIV) infection and multiple drug exposures, including fluconazole (400 mg/day) until the fifth month and then from 6 months to term, efavirenz, nevirapine, methadone, dapsone, pentamidine, and trimethoprim sulfamethoxazole. At birth the infant had seizures related to neonatal abstinence syndrome and was noted to have multiple congenital anomalies. On examination at age 9 months, he had craniosynostosis secondary to coronal and lambdoidal suture closures, shallow orbital region, hypoplastic supraorbital ridges, hypertelorism, and mild ptosis. He had radioulnar synostosis and metacarpophalangeal-proximal interphalangeal symphalangism of D2-D5 bilaterally. CONCLUSIONS: The findings of cranial synostosis, multiple symphalangism, and long-bone abnormalities in our case are typical of other reported cases of fluconazole embryopathy. Our patient showed no evidence of embryopathy due to efavirenz, and he did not have the features of Antley-Bixler or other craniosynostosis syndromes. We review the literature regarding the teratogenic effects of prenatal exposure to fluconazole and provide additional evidence that prenatal fluconazole exposure has a clearly identifiable phenotype. PMID- 16265640 TI - Marshall J. Edwards: discoverer of maternal hyperthermia as a human teratogen. AB - In a series of animal studies performed over a career spanning 40 years at the University of Sydney, Professor Marshall J. Edwards investigated the hypothesis that maternal hyperthermia during gestation can be teratogenic to the developing fetus. He is one of few investigators to have discovered a known human teratogen primarily through animal studies. In 1970 he earned his Ph.D. from the University of Sydney, writing a doctoral thesis entitled "A Study of Some Factors Affecting Fertility of Animals with Particular Reference to the Effects of Hyperthermia on Gestation and Prenatal Development of the Guinea-Pig." He went on to prove that hyperthermia-induced malformations in animals involve many organs and structures, particularly the central nervous system. Other defects include craniofacial anomalies, heart defects and hypodactyly, cataracts and coloboma, kyphoscoliosis, renal anomalies, dental agenesis, and abdominal wall defects. In a series of carefully planned and executed experiments, he demonstrated that the type of defect is related to the timing of the hyperthermic insult, and analyzed the underlying mechanisms. Cell death, membrane disruption, vascular disruption, and placental infarction were all implicated in causing embryonic damage. This special article reviews the scientific discoveries and personal philosophy of Marshall J. Edwards, the discoverer of maternal hyperthermia as a human teratogen. PMID- 16265641 TI - Sex differences in birth defects: a study of opposite-sex twins. AB - BACKGROUND: Sex differences in structural birth defects are often confounded by environmental risk factors. Opposite-sex twins provide a unique model for detecting sex differences in birth defects while maximally controlling environmental risk factors in a natural setting. METHODS: Population data from the Florida Birth Defects Registry were analyzed. A total of 4,768 pairs of twins who were discordant for sex and born between 1996 and 2001 were analyzed. The McNemar test was used to compare the differences between a male twin and his twin sister for the risk of developing specific defects and organ-system defects. RESULTS: Of 4,768 twin pairs, 225 males (4.72%) and 175 females (3.67%) had birth defects. Among opposite-sex twin pairs, males had a 29% higher risk for birth defects than their twin sisters. Compared to their twin sisters, males had a 5.4 times higher risk for pyloric stenosis and a 2.4 times higher risk for obstructive genitourinary defect, but only one-tenth the risk for congenital hip dislocation. CONCLUSIONS: Sex differences in birth defects exist between opposite sex twins. PMID- 16265642 TI - Inference of multiple fiber orientations in high angular resolution diffusion imaging. AB - A method is presented that is capable of determining more than one fiber orientation within a single voxel in high angular resolution diffusion imaging (HARDI) data sets. This method is an extension of the Markov chain method recently introduced to diffusion tensor imaging (DTI) analysis, allowing the probability density function of up to 2 intra-voxel fiber orientations to be inferred. The multiple fiber architecture within a voxel is then assessed by calculating the relative probabilities of a 1 and 2 fiber model. It is demonstrated that for realistic signal to noise ratios, it is possible to accurately characterize the directions of 2 intersecting fibers using a 2 fiber model. The shortcomings of under-fitting a 2 fiber model, or over-fitting a 1 fiber model, are explored. This new algorithm enhances the tools available for fiber tracking. PMID- 16265643 TI - Quantitative assessment of coronary artery plaque vulnerability by high resolution magnetic resonance imaging and computational biomechanics: a pilot study ex vivo. AB - The risk of atherosclerotic plaque disruption is thought to be closely related to plaque composition and rupture triggers such as external mechanical forces. The purpose of this study was to integrate MR imaging and computational techniques for the quantification of plaque vulnerability with morphologic data and biomechanical stress/strain distributions that were all based on high-resolution MR images of coronary artery plaque specimens ex vivo. Twenty-two coronary artery plaque specimens were selectively collected from 10 cadavers. Multislice T(2) weighted spin echo images were acquired with a resolution of 100 x 100 microm(2). Histopathological images were used as the gold standard for the identification of plaque components and vulnerability. Plaque components were classified on MR images, and the stress/strain components were calculated with a two-dimensional computational model with fluid-structure interactions. As expected, vulnerable plaques appeared to result from a large lipid pool, a thin fibrous cap, and some critical stress/strain conditions. An empiric vulnerability marker was derived and was closely related to the vulnerability score that was determined through pathologic examination. Noninvasive quantification of the MR contrast and mechanical properties of plaque may provide a comprehensive biomarker for the assessment of vulnerability of atherosclerotic plaques. PMID- 16265644 TI - Dependence on diffusion time of apparent diffusion tensor of ex vivo calf tongue and heart. AB - The time dependence of the apparent diffusion tensor of ex vivo calf heart and tongue was measured for diffusion times (tau(d)) between 32 and 810 ms. The results showed evidence of restricted diffusion in the muscle tissues of both organs. In regions where the myofibers are parallel, the largest eigenvalue (lambda(1)) of the diffusion tensor remained the same for all diffusion times measured, while the other eigenvalues (lambda(2), lambda(3)) decreased by 29-36% between tau(d) = 32 ms and tau(d) = 400 ms. In regions where the fibers cross, the lambda(1) also changed, decreasing by 17% between tau(d) = 32 ms and tau(d) = 400 ms. The restricting compartment size and volume fraction were effectively estimated by fitting the time courses of the eigenvalues to a model consisting of a nonrestricted compartment and a cylindrically restricted compartment. To our knowledge, this study is the first demonstrating diffusion time dependence of measured water diffusion tensor in muscular tissue. With improvement in scanning technology, future studies may permit noninvasive, in vivo detection of changes in muscle myoarchitecture due to disease, treatment, and exercise. PMID- 16265645 TI - Morphological features of ovine embryonic lung fibroblasts cultured on different bioactive scaffolds. AB - Tissue regeneration with autologous cell transplantation is one of the most important goals in clinical research. In this field, the development of bioactive materials that provide microenvironments for cell-matrix interactions mimicking biological conditions is required. In recent years, many synthetic materials have been developed as scaffolds and many procedures for the surface modification of these materials have been applied using biological molecules. In this study, we analyzed the morphology and the molecule production by ovine embryonic lung fibroblasts cultured on three different sponge-like matrices based on poly(L lactic acid) (PLLA): agarose/PLLA, crosslinked and uncrosslinked gelatin/PLLA. The matrices were produced by using an emulsion freeze-drying method leading to the formation of sponge-like materials with high porosity and with interconnection between the pores. In vitro MTT test demonstrated that transplanted cells were viable and metabolically active. Morphological analysis revealed that fibroblasts adhered to and penetrated the polymeric structures. Moreover, all the different matrices supported fibroblast production of proteoglycans, glycoproteins, and matrix molecules such as elastin, collagen I, and fibronectin. These data suggest that the tested bioactive scaffolds may support the growth and extracellular matrix molecule production of fibroblasts allowing in vitro connective tissue regeneration. PMID- 16265646 TI - Gene-environment interactions in rare diseases that include common birth defects. AB - Rare syndromes often feature specific types of birth defects that frequently are major diagnostic clues to the presence of a given disorder. Despite this specificity, not everyone with the same syndrome is equally or comparably affected, and not everyone with a specific birth defect manifests the same syndrome or is affected with all the features of a particular syndrome. A symposium sponsored by the National Institutes of Health Office of Rare Diseases, and the National Toxicology Program Center for the Evaluation of Risks to Human Reproduction attempted to explore how much of this variability is due to genetic factors and how much is due to environmental factors. The specific types of birth defects examined included cardiovascular defects, holoprosencephaly, clefts of the lip and/or palate, neural tube defects, and diaphragmatic hernias. PMID- 16265647 TI - Evaluation of alginate purification methods: effect on polyphenol, endotoxin, and protein contamination. AB - Alginate, a polysaccharide extracted from brown seaweed, is widely used for the microencapsulation of islets of Langerhans, allowing their transplantation without immunosuppression. This natural polymer is known to be largely contaminated. The implantation of islets encapsulated using unpurified alginate leads to the development of fibrotic cell overgrowth around the microcapsules and normalization of the blood glucose is restricted to a very short period if it is achieved at all. Several research groups have developed their own purification method and obtained relatively good results. No comparative evaluation of the efficiencies of these methods has been published. We conducted an evaluative study of five different alginate preparations: a pharmaceutical-grade alginate in its raw state, the same alginate after purification according to three different published methods, and a commercially available purified alginate. The results showed that all purification methods reduced the amounts of known contaminants, that is, polyphenols, endotoxins, and proteins, although with varying efficiencies. Increased viscosity of alginate solutions was observed after purification of the alginates. Despite a general efficiency in decreasing contamination levels, all of the purified alginates contained relatively high residual amounts of protein contaminants. Because proteins may be immunogenic, these residual proteins may have a role in persisting microcapsule immunogenicity. PMID- 16265648 TI - "Wet-state" mechanical properties of three-dimensional polyester porous scaffolds. AB - Porous poly(D,L-lactic-co-glycolic acid) (PLGA) scaffolds under a simulated physiological environment were investigated to estimate their "true" mechanical properties, with emphasis on the effect of "wet-state" on the compressive behaviors. The effect of the history of ethanol sterilization was also investigated. The studies were focused upon the "wet-state" mechanical properties of polyester porous scaffolds, because the potential implants must be used under a wet environment. The measurements of three-dimensional porous scaffolds composed of amorphous PLGA with five polymer formulations including poly(D,L lactic acid) (PDLLA) demonstrated that the mechanical properties of PLGA scaffolds significantly decreased in phosphate buffer saline solution (PBS) at 37 degrees C and/or with an ethanol sterilization history, even though PLGA is a hydrophobic material. The decrease extent depends on the copolymer composition: when the porosity is about 90%, a PDLLA scaffold remained about 75-80% of initial mechanical properties in the dry state at 25 degrees C, whereas PLGA 85:15, 75:25, and 65:35 scaffolds remained only about 10% or less, and the PLGA 50:50 scaffolds examined were not sufficiently strong for mechanical tests. If scaffolds were prewetted with ethanol ahead of prewetting with PBS, the mechanical properties further decreased compared with those merely prewetted with PBS. These phenomena were elucidated experimentally from plasticization of PLGA with water or ethanol, and the consequent reduction of glass transition temperature. The results might be helpful for designing polyester porous scaffolds for tissue engineering or in situ tissue induction applications. PMID- 16265649 TI - Mesenchymal stem cell adhesion and spreading on microwave plasma-nitrided titanium alloy. AB - Improved methods to increase surface hardness of metallic biomedical implants are being developed in an effort to minimize the formation of wear debris particles that cause local pain and inflammation. However, for many implant surface treatments, there is a risk of film delamination due to the mismatch of mechanical properties between the hard surface and the softer underlying metal. In this article, we describe the surface modification of titanium alloy (Ti-6Al 4V), using microwave plasma chemical vapor deposition to induce titanium nitride formation by nitrogen diffusion. The result is a gradual transition from a titanium nitride surface to the bulk titanium alloy, without a sharp interface that could otherwise lead to delamination. We demonstrate that vitronectin adsorption, as well as the adhesion and spreading of human mesenchymal stem cells to plasma-nitrided titanium is equivalent to that of Ti-6Al-4V, while hardness is improved 3- to 4-fold. These in vitro results suggest that the plasma nitriding technique has the potential to reduce wear, and the resulting debris particle release, of biomedical implants without compromising osseointegration; thus, minimizing the possibility of implant loosening over time. PMID- 16265651 TI - Surface modification and initial adhesion events for intestinal epithelial cells. AB - Rapid resealing of the mucosal epithelia is imperative following injuries to the small intestine because the mucosa is responsible for the adsorption of nutrients as well as providing a barrier to noxious agents present in the lumen. Tissue engineering may provide a possible solution for treating intestinal erosions, ulcerations, inflammatory bowel disease, and infection. Cell-biomaterial interaction is a critical component in tissue engineering that can determine the success of the tissue construct. Cell-biomaterial interactions can be enhanced by various types of surface modification, which promote integrin ligation leading to increased cell function. In order to relate the effect of surface adhesion molecules to signaling events and macroscopic cell response, an intestinal epithelial cell line, IEC-6, was plated on fibronectin (receptor-mediated) and poly-L-lysine (non-specific) surfaces. Focal adhesion kinase (FAK) phosphorylation, cell spreading, and cell adhesion strength were measured. Results showed increases in FAK phosphorylation generally corresponded to increases in cell spreading and adhesion strength for IEC-6 cells. Therefore, in a simplified system, initial adhesion and signaling mechanisms appeared to correspond to subsequent physical responses in IEC-6 cells relevant to tissue engineering applications. PMID- 16265650 TI - Dexamethasone-functionalized gels induce osteogenic differentiation of encapsulated hMSCs. AB - Synthetic hydrogels represent highly controlled environments for three dimensional culture of human mesenchymal stem cells (hMSCs). Encapsulated hMSCs are presented with a "blank" environment, and this environment can be closely controlled in order to elicit an osteogenic response. In vitro, dexamethasone is an efficient and reliable factor that leads to the osteogenic differentiation of human mesenchymal stem cells (hMSCs). The aim of this work was to develop a dexamethasone-releasing poly(ethylene glycol) (PEG)-based hydrogel scaffold to deliver dexamethasone to encapsulated cells in a sustained manner. To accomplish this goal, dexamethasone was covalently linked to a photoreactive mono-acrylated PEG molecule through a degradable lactide bond, and this molecule was covalently incorporated into the PEG hydrogel during photopolymerization. Over time, hydrolysis of the ester bonds resulted in dexamethasone release from the gel. The biological activity of the released dexamethasone was verified in monolayer cell culture and in three-dimensional culture (i.e., in the gel) by the ability of hMSCs to express osteogenic genes, including alkaline phosphatase, osteopontin, and core binding factor alpha 1, as measured using real-time reverse transcription polymerase chain reaction (RT-PCR). These studies indicate that encapsulated hMSCs are capable of osteogenic differentiation in response to released dexamethasone. PMID- 16265652 TI - Metalloproteinases and their inhibitors: influence on tumor invasiveness and metastasis formation in head and neck squamous cell carcinomas. AB - BACKGROUND: Matrix metalloproteinases (MMPs) play an important role in tumor invasiveness. This study investigates the expression status of MMPs and tissue inhibitors of metalloproteinases (TIMPs) in head and neck squamous cell carcinomas (HNSCC). METHODS: Of 48 laryngeal squamous cell carcinoma (LSCC) biopsies and 10 HNSCC cell lines, mRNA was isolated, reversely transcribed, and subjected to polymerase chain reaction (PCR) amplifying MMP-1, MMP-2, MMP-9, MMP 10, TIMP-1, and TIMP-2. Silver nitrate-stained gel electrophoresis demonstrated MMP and TIMP expression status. Exemplary immunohistochemistry and zymography confirmed translation and enzyme activity. RESULTS: Densitometric analysis revealed MMP-2 expression and lymph node metastases to be positively and TIMP-1 and TIMP-2 to be negatively correlated with lymph node metastases. TIMP-2 expression and tumor size were negatively correlated. MMP-1, MMP-9, and MMP-10 expression were not correlated to metastasis formation or tumor size. CONCLUSIONS: Our results suggest that MMP-2 expression enhances, whereas TIMP-1 and TIMP-2 both suppress, cancer spread in LSCC. PMID- 16265653 TI - Clinical trials in head and neck oncology: an evaluation of clinicians' willingness to participate. AB - BACKGROUND: This study investigated the individual and collective ("community") equipoise of surgeons and oncologists and their willingness to take part in each of six hypothetical randomized controlled trials in head and neck oncology. METHODS: A survey was mailed to Australasian head and neck specialists. RESULTS: Among 109 respondents (74% response), the scenario with the highest level of individual equipoise pertained to the use of adjuvant interferon for patients with high-risk malignant melanoma, with 45% indicating complete uncertainty between treatment approaches. Significant differences in levels of community equipoise were demonstrated between surgeons and oncologists for three of the scenarios. Willingness to participate in randomized controlled trials ranged from 39% to 72%. Increasing strength of treatment preference was associated with unwillingness to participate in randomized controlled trials for two of six scenarios. CONCLUSION: High levels of equipoise and willingness to participate in clinical research augur well for future randomized controlled trials in head and neck oncology. PMID- 16265654 TI - Cervicofacial and cervicothoracic rotation flaps in head and neck reconstruction. AB - BACKGROUND: Compound cervicofacial and cervicothoracic rotation flaps are highly versatile flaps that may be applied to a variety of defects of the cheek, orbit, periauricular region, and neck. These rotation advancement flaps should be a staple of the head and neck surgeon's reconstructive armamentarium. METHODS: This is a retrospective review of medical records at a university-based head and neck cancer center. RESULTS: Thirty-three patients were identified, with a mean age of 66 years. Primary or recurrent skin neoplasms made up the most common indication for surgery, followed by primary parotid tumors and cervical lymphatic metastases from upper aerodigestive tract malignancies. Defects of the cheek, orbit, periauricular region, and neck were reconstructed with cervicofacial or cervicothoracic flaps, with larger wounds requiring variable extension of the incision onto the chest wall. Other reconstructive modalities were used in 18 cases to increase tissue bulk or provide internal lining. Minor wound complications occurred in 13 patients. There was no statistically significant association between wound complications and smoking or previous radiation therapy. CONCLUSIONS: Compound cervicofacial and cervicothoracic rotation flaps provide a straightforward, reliable, and efficient means to reconstruct complex defects of the face, lateral skull base, and neck, with the potential for excellent cosmetic results. PMID- 16265655 TI - Unexpected rapid progression of metastatic adenoid cystic carcinoma during treatment with imatinib mesylate. AB - BACKGROUND: There is a lack of effective treatment for metastatic adenoid cystic carcinoma (ACC), a usually indolent tumor. We studied the efficacy of imatinib mesylate, a potent inhibitor of KIT tyrosine kinase, in patients with KIT positive metastatic ACC. METHOD: Five patients with lung metastasis were treated in a pilot study with imatinib 400 mg by mouth twice a day. Mutations of c-kit and platelet-derived growth factor receptor (PDGFR)-alpha in tumors from these patients were analyzed. RESULTS: Disease progression was noted in three of five patients during the short treatment periods, ranging from 2 to 3 weeks. Three patients died of disease within 6 months. No detectable mutations were found in c kit and PDGFR-alpha. CONCLUSION: We observed an unexpected high progression rate of metastatic ACC within short periods during imatinib treatment. Use of imatinib to treat cancers without c-kit or PDGFR-alpha mutation should be approached with caution. PMID- 16265656 TI - 25-year experience of using a linear stapler in laryngectomy. AB - BACKGROUND: Stapler application for pharyngeal closure after total laryngectomy allows for rapid watertight closure without field contamination and for potentially reduced fistula rate. METHODS: One thousand four hundred fifteen patients underwent laryngectomy with linear stapler closure. In 98.6%, laryngectomy was performed after radiation failure. RESULTS: A relatively high incidence of pharyngeal fistulae (12%) was seen, although these rates were reduced to 5.5% during the recent decade. Simultaneous creation of tracheoesophageal fistula and myotomy by a novel technique was introduced. Swallowing problems were observed in 11 patients and local recurrences in nine patients (0.6%). CONCLUSION: The advantages of mechanical sutures with the closed stapling technique are simple and rapid application, watertight closure with good hemostasis, prevention of field contamination, good speech and deglutition, no increase in fistula rate, and low local recurrence rates. Operating room expenses may also be significantly reduced, rendering this method cost-effective as well. PMID- 16265657 TI - Second primary malignancy of the aerodigestive tract in patients treated for cancer of the oral cavity and larynx. AB - BACKGROUND: We aimed to identify patterns and predictors of second primary malignancy (SPM) of the aerodigestive tract (SPMADT) in patients with squamous cell carcinoma of the oral cavity (SCCOC) and larynx (SCCL). METHODS: One thousand two hundred fifty-seven patients from two existing databases were studied: 595 with SCCOC (1986-1995) and 662 with SCCL (1984-1998). The primary endpoint of interest was development of SPMADT, defined as a second primary neoplasm of the head and neck, esophagus, or lung. RESULTS: The 5-year SPMADT rate was 8% in the SCCL versus 10% in the SCCOC subgroup. Lung SPM was more common in the SCCL group; head and neck SPM was more common in the SCCOC group. Smokers had a fivefold increased risk, whereas alcohol use was associated with a twofold increased risk of SPMADT. CONCLUSIONS: The rates of SPMADT after treatment of SCCOC and SCCL are comparable, but the patterns are different. Smoking and alcohol use are independent predictors of SPMADT development. PMID- 16265659 TI - Q-switched ruby versus long-pulsed dye laser delivered with compression for treatment of facial lentigines in Asians. AB - BACKGROUND AND OBJECTIVES: Q-switched lasers have been used for the treatment of lentigines but post-inflammatory hyperpigmentation (PIH) can be an issue especially in Asians. The 595 nm long-pulsed dye laser (LPDL) has been used for the treatment of vascular lesions and although it is well absorbed by oxyhemoglobin, it is also absorbed by melanin. To use this device for the treatment of facial lentigines, we attached a flat glass lens to the tip of the laser's handpiece, allowing compression of the skin during treatment. In doing so, eliminated the absorption by oxyhemoglobin. This prospective study aims to compare the efficacy and complications of such an approach to the use of Q switched ruby laser (QSRL) in the treatment of facial lentigines in Asians. STUDY DESIGN/MATERIALS AND METHODS: Eighteen Asian patients (1 male, 17 female) with facial lentigines Fitzpatrick skin types III-IV were enrolled. One of the lentigines present was treated with LPDL by compression method and the other one was treated with QSRL. A LPDL emitting wavelength of 595 nm, spot size of 7 mm was used, with fluence between 10 and 13 J/cm(2) and pulse duration of 1.5 milliseconds. Cryogen spray cooling was not used. A 694 nm QSRL was used with a spot size of 4 mm, fluence of 6-7 J/cm(2), and pulse duration of 30 nanoseconds. Lightening of the lesions was assessed by reflectance spectrometer Erythema, hypo or hyperpigmentation and scarring were also assessed by clinical examinators. RESULTS: The degree of clearing achieved with the two lasers was 70.3% and 83.3% for QSRL and LPDL, respectively. All QSRL treated areas developed erythema whereas only 4 of 18 LPDL treated areas developed erythema. Hyperpigmentation was seen in four patients after QSRL, but not after LPDL. There was no scarring or hypopigmentation. CONCLUSIONS: LPDL delivered with a compression method is more effective than QSRL for facial lentigines. Complications after LPDL treatment were substantially less frequent than after QSRL. The addition of compression technique may allow "vascular" pulsed dye laser to be used for treating a variety of pigmented lesions. PMID- 16265658 TI - Concomitant low-dose cisplatin and three-dimensional conformal radiotherapy for locally advanced squamous cell carcinoma of the head and neck: Analysis of survival and toxicity. AB - BACKGROUND: Our center sought to implement a simple chemoradiotherapy schedule for patients with locally advanced squamous cell carcinoma of the head and neck (SCCHN) with minimal toxicity to achieve rates of overall survival comparable to other schedules. METHODS: The chemoradiotherapy schedule consisted of daily radiation to 70 Gy over 7 weeks with concurrent cisplatin 20 mg/m(2) during days 1 to 4 of weeks 1 and 5. Acute and late toxicities were recorded according to the Radiation Therapy Oncology Group (RTOG) and common toxicity criteria (CTC) grading. The overall, disease-specific, and locoregional recurrence-free survival were calculated using the STATA statistics package. Possible factors influencing these endpoints were analyzed. RESULTS: Fifty-seven patients were treated, and 56 patients were evaluable for follow-up. Median follow-up of alive patients was 16.1 months. There was an 82% complete response rate to chemoradiotherapy. The 2 year Kaplan-Meier overall, disease-specific, and locoregional recurrence-free survival rates were 62%, 67%, and 63%. Acute grade 3 and 4 radiation toxicity was noted in 61% and 2%, respectively. Grade 3 or 4 hematologic toxicity was noted in 7% of patients. Factors influencing overall survival included: Karnofsky performance status, receiving more than 50% of planned chemotherapy, age, and initial hemoglobin level. CONCLUSION: This regimen is tolerable and achieves overall survival and locoregional control rates comparable to other chemoradiotherapy schedules. PMID- 16265660 TI - Ciprofloxacin-releasing bioabsorbable polymer is superior to titanium in preventing Staphylococcus epidermidis attachment and biofilm formation in vitro. AB - Antibiotic coating systems have been successfully used to prevent bacterial attachment and biofilm formation. Our purpose was to evaluate whether bioabsorbable polylactide-co-glycolide (PLGA) 80/20 on its own, and PLGA together with ciprofloxacin (PLGA+C) have any advantages over titanium in preventing Staphylococcus epidermidis attachment and biofilm formation in vitro. Cylindrical specimens of titanium, PLGA, and PLGA+C in triplicate were examined for S. epidermidis ATCC 35989 attachment and biofilm formation after incubation with a bacterial suspension of about 10(5) cfu/mL for 1, 3, 7, 14, and 21 days, using scanning electron microscopy. Growth inhibition properties of PLGA and PLGA+C cylinders were tested on agar plates. On days 1, 3, and 21, no bacterial attachment was seen in 19.5, 9.2, and 41.4% of the titanium specimens; in 18.4, 28.7, and 34.5% of the PLGA specimens; and in 57.5, 62.1, and 57.5% of the PLGA+C specimens, respectively. During the whole study period, no biofilm was observed on 74-93% of the titanium specimens, 58-78% of the PLGA specimens, and 93-100% of the PLGA+C specimens. PLGA+C showed clear bacterial growth inhibition on agar plates, while PLGA and titanium did not show any inhibition. PLGA+C bioabsorbable material was superior to titanium in preventing bacterial attachment and biofilm formation and may have clinical applicability, for example, in prevention of infection in trauma surgery or in the treatment of chronic osteomyelitis. PMID- 16265661 TI - Sensitivity to implant materials in patients undergoing total hip replacement. AB - Sensitivity to implant materials is an unpredictable event, which may contribute to the process leading to the failure of the total hip replacement (THR). The aim of the current study was to investigate the informative power of skin testing in detecting the sensitization to the implant components in patients undergoing THR. A consecutive series of 223 patients was enrolled in the study, including 66 candidates to THR, 53 with stable implant, and 104 with THR loosening. The patch testing was performed by using the most relevant components of cobalt-based alloys (CoCrMo), Ti-based alloys (TiAlV), and bone cements. The frequency of positive patch testing in preimplant patients did not differ from that observed after THR. Patients with CoCrMo-failed implant showed a significant low frequency of nickel-positive skin reaction, while patients with TiAlV-THR had a high incidence of vanadium-positive patch testing. The panel of haptens showed a good performance in the identification of patients known to have a contact dermatitis. With regard to the THR outcome, patch testing was not able to discriminate between stable and failed implant. Sensitivity to at least one hapten, namely bone cement, as well as the positive medical history of hypersensitivity, influenced negatively the THR survival. Our results show the reliability of patch testing for investigating the sensitivity to implant components. The cause-effect relationship between sensitization and negative outcome cannot be established, but the shorter lifespan of THR in patients who have a positive patch testing supports the significant role of this event in contributing to the implant failure. PMID- 16265663 TI - Recombinant human deoxyribonuclease shortens ventilation time in young, mechanically ventilated children. AB - Recombinant human deoxyribonuclease I (dornase alfa) is currently used as an inhaled mucoactive agent in the treatment of cystic fibrosis. In a randomized, placebo-controlled, double-blind clinical study in 100 infants, we investigated whether the therapeutic use of dornase alfa can be extended to ventilated, fluid restricted children to reduce reintubation rate, ventilation duration, pediatric intensive care unit (PICU) stay, and ventilation complications. While reintubation rates were similar for dornase alfa 7% vs. placebo 9% (odds ratio, 0.77; confidence interval, 0.11-4.9), the incidence of atelectasis (6 vs. 17, respectively; P-value 0.051), median ventilation time (2.2 vs. 3.4 days, respectively; P-value 0.043), median length of PICU stay (7 vs. 8 days, respectively; P-value 0.051), and mean costs (4,830 vs. 6,320, respectively) were lower in the dornase alfa group. No adverse effects were observed, even in critically ill patients. We found that dornase alfa was beneficial and safe. Our findings also indicate that dornase alfa is possibly of value from the first day of mechanical ventilation onward, particularly when longer ventilation (>3 days) is expected in fluid-restricted children after cardiac surgery. PMID- 16265662 TI - Local release of dexamethasone from polymer millirods effectively prevents fibrosis after radiofrequency ablation. AB - Recent studies show that after radiofrequency (RF) ablation, fibrosis occurs at the ablation boundary, hindering anticancer drug transport from a locally implanted polymer depot to the ablation margin, where tumors recur. The purpose of this study is to investigate strategies that can effectively deliver dexamethasone (DEX), an anti-inflammatory agent, to prevent fibrosis. Polymer millirods consisting of poly(D,L-lactide-co-glycolide) (PLGA) were loaded with either DEX complexed with hydroxypropyl beta-cyclodextrin (HPbeta-CD), or an NaCl and DEX mixture. In vitro release studies show that DEX complexed with HPbeta-CD released 95% of the drug after 4 days, compared to 14% from millirods containing NaCl and DEX. Rat livers underwent RF ablation and received either DEX-HPbeta-CD loaded millirods, PLGA millirods with an intraperitoneal (i.p.) DEX injection, or control PLGA millirods alone. After 8 days in vivo, heightened inflammation and the appearance of a well-defined fibrous capsule can be observed in both the control experiments and those receiving a DEX injection (0.29 +/- 0.08 and 0.26 +/- 0.07 mm in thickness, respectively), with minimal inflammation and fibrosis present in livers receiving DEX millirods (0.04 +/- 0.01 mm). Results from this study show that local release of DEX prevents fibrosis more effectively than a systemic i.p. injection. PMID- 16265664 TI - The transcriptional regulation role of BRD7 by binding to acetylated histone through bromodomain. AB - Studies showed that the bromodomain binds to acetyl-lysines on histone tails, which is involved in deciphering the histone codes. BRD7, a novel bromodomain gene, is the first described bromodomain gene involved in nasopharyngeal carcinoma (NPC). Previous studies showed that ectopic expression of BRD7 inhibited cell growth and cell cycle progression from G1 to S phase in HNE1 cells (a NPC cell line) by transcriptionally regulating some cell cycle related genes including E2F3 gene. In the present study, we revealed the co-localization between acetylated H3 and BRD7 and found that the bromodomain of BRD7 is required for this co-localization. More importantly, wild-type BRD7 interacted with H3 peptide acetylated at Lys14, while the bromodomain deleted mutant lost this ability. We also found that the mutant BRD7 failed to regulate E2F3 promoter activity and inhibit cell cycle progression. These results indicated that the transcriptional regulation role of BRD7 was achieved by binding to acetylated histone H3 and that the bromodomain was essential for this role. In addition, no obvious changes were observed in the acetylated level of histone H3 after transfection with BRD7, indicating that chromatin remodeling, not chromatin modification, is the major mechanism of BRD7 mediated gene transcription. Taken together, the present work shed light on the fact that a novel bromodomain gene, BRD7, is of importance in transcriptional regulation and cellular events including cell cycle. PMID- 16265665 TI - Cyclin D1 gene numerical aberration is a predictive marker for occult cervical lymph node metastasis in TNM Stage I and II squamous cell carcinoma of the oral cavity. AB - BACKGROUND: The management of occult cervical lymph node metastases originating from oral squamous cell carcinomas (OSCCs) remains controversial. The purpose of this study was to evaluate the value of cyclin D1 gene (CCND1) numerical aberrations in predicting the risk of late lymph node metastases. METHODS: Fluorescence in situ hybridization (FISH), using a BAC clone specific for CCND1, was performed on OSCC specimens obtained by fine-needle aspiration (FNA) biopsy from 45 patients with previously untreated TNM Stage I and II (T1-2N0M0) disease who had not undergone elective cervical lymph node dissection. RESULTS: CCND1 numerical aberrations were observed in 15 (33.3%) of the 45 patients and were significantly associated with the mode of invasion of the primary tumor (P = 0.01) and the presence of occult lymph node metastases (P < 0.001). Twelve of these 15 patients (80%) developed late cervical lymph node metastases within 2 years of surgery for primary OSCCs. All patients with cluster-type amplification of CCND1 developed late lymph node metastases. Multivariate analysis showed that only CCND1 numerical aberrations (risk ratio, 8.685%, 95% confidence interval = 2.232-33.802, P = 0.002) independently predicted late cervical lymph node metastasis. CONCLUSIONS: Aberrations in CCND1 numbers appear to be valuable in identifying patients at high risk of late lymph node metastasis in Stage I and II OSCCs. Analysis of CCND1 numerical aberrations using FISH on FNA biopsy specimens may be useful in selecting patients for elective cervical lymph node dissection. PMID- 16265666 TI - Functional demonstration of surface carbonic anhydrase IV activity on rat astrocytes. AB - Buffering of the brain extracellular fluid is catalyzed by carbonic anhydrase (CA) activity. Whereas the extracellular isoform CA XIV has been localized exclusively to neurons in the brain, and to glial cells in the retina, there has been uncertainty regarding the form or forms of CA on the surface of brain astrocytes. We addressed this issue using physiological methods on cultured and acutely dissociated rat astrocytes. Prior work showed that the intracellular lactate-induced acidification (LIA) of astrocytes is diminished by benzolamide, a poorly permeant, nonspecific CA inhibitor. We demonstrate that pretreatment of astrocytes with phosphatidylinositol-specific phospholipase C (PI-PLC) results in a similar inhibition of the mean LIA (by 66 +/- 3%), suggesting that the glycosylphosphatidylinositol-anchored CA IV was responsible. Pretreatment of astrocytes with CA IV inhibitory antisera also markedly reduced the mean LIA in both cultured cortical (by 46 +/- 4%) and acutely dissociated hippocampal astrocytes (by 54 +/- 8%). Pre-immune sera had no effect. The inhibition produced by PIPLC or CA IV antisera was not significantly less than that by benzolamide, suggesting that the majority of detectable surface CA activity was attributable to CA IV. Thus, our data collectively document the presence of CAIV on the surface of brain astrocytes, and suggest that this is the predominant CA isoform on these cells. PMID- 16265667 TI - TLR3-mediated signal induces proinflammatory cytokine and chemokine gene expression in astrocytes: differential signaling mechanisms of TLR3-induced IP-10 and IL-8 gene expression. AB - Viral infection is one of the leading causes of brain encephalitis and meningitis. Recently, it was reported that Toll-like receptor-3 (TLR3) induces a double-stranded RNA (dsRNA)-mediated inflammatory signal in the cells of the innate immune system, and studies suggested that dsRNA may induce inflammation in the central nervous system (CNS) by activating the CNS-resident glial cells. To explore further the connection between dsRNA and inflammation in the CNS, we have studied the effects of dsRNA stimulation in astrocytes. Our results show that the injection of polyinosinic-polycytidylic acid (poly(I:C)), a synthetic dsRNA, into the striatum of the mouse brain induces the activation of astrocytes and the expression of TNF-alpha, IFN-beta, and IP-10. Stimulation with poly(I:C) also induces the expression of these proinflammatory genes in primary astrocytes and in CRT-MG, a human astrocyte cell line. Furthermore, our studies on the intracellular signaling pathways reveal that poly(I:C) stimulation activates IkappaB kinase (IKK), extracellular signal-regulated kinase (ERK), and c-Jun N terminal kinase (JNK) in CRT-MG. Pharmacological inhibitors of nuclear factor kappaB (NF-kappaB), JNK, ERK, glycogen synthase kinase-3beta (GSK-3beta), and dsRNA-activated protein kinase (PKR) inhibit the expression of IL-8 and IP-10 in astrocytes, indicating that the activation of these signaling molecules is required for the TLR3-mediated chemokine gene induction. Interestingly, the inhibition of PI3 kinase suppressed the expression of IP-10, but upregulated the expression of IL-8, suggesting differential roles for PI3 kinase, depending on the target genes. These data suggest that the TLR3 expressed on astrocytes may initiate an inflammatory response upon viral infection in the CNS. PMID- 16265668 TI - His36Pro point-mutated proteolipid protein retained in the endoplasmic reticulum of oligodendrocytes in the shaking pup. AB - The shaking pup (shp) is a canine mutation that affects the myelin protein proteolipid protein (PLP) and its smaller and less abundant isoform, DM20, with proline replacing histidine(36), resulting in a severe myelin deficiency in the central nervous system. We present evidence that the mutation leads to disrupted trafficking of the shp PLP/DM20 within oligodendrocytes. Immunohistochemical studies revealed significantly reduced levels of PLP/DM20 and other major myelin components such as myelin basic protein (MBP), myelin associated glycoprotein (MAG), and 2',3'-cyclic nucleotide 3'-phosphodiesterase (CNP) in shp myelin. The distribution of shp PLP/DM20 proteins were altered and mostly retained in perinuclear cytoplasm and proximal processes, which co-localized with distended rough endoplasmic reticulum (RER) within oligodendrocytes. No abnormal accumulation of MAG, MBP, or CNP in the cell body was found. These results suggest that mutated PLP/DM20 in the shp could be selectively retained in RER, causing disruption of their translocation to the periphery to myelinate axons. PMID- 16265669 TI - Tandem-pore domain potassium channels are functionally expressed in retinal (Muller) glial cells. AB - Tandem-pore domain (2P-domain) K+-channels regulate neuronal excitability, but their function in glia, particularly, in retinal glial cells, is unclear. We have previously demonstrated the immunocytochemical localization of the 2P-domain K+ channels TASK-1 and TASK-2 in retinal Muller glial cells of amphibians. The purpose of the present study was to determine whether these channels were functional, by employing whole-cell recording from frog and mammalian (guinea pig, rat and mouse) Muller cells and confocal microscopy to monitor swelling in rat Muller cells. TASK-like immunolabel was localized in these cells. The currents mediated by 2P-domain channels were studied in isolation after blocking Kir, K(A), K(D), and BK channels. The remaining cell conductance was mostly outward and was depressed by acid pH, bupivacaine, methanandamide, quinine, and clofilium, and activated by alkaline pH in a manner consistent with that described for TASK channels. Arachidonic acid (an activator of TREK channels) had no effect on this conductance. Blockade of the conductance with bupivacaine depolarized the Muller cell membrane potential by about 50%. In slices of the rat retina, adenosine inhibited osmotic glial cell swelling via activation of A1 receptors and subsequent opening of 2P-domain K+ channels. The swelling was strongly increased by clofilium and quinine (inhibitors of 2P-domain K+ channels). These data suggest that 2P-domain K+ channels are involved in homeostasis of glial cell volume, in activity-dependent spatial K+ buffering and may play a role in maintenance of a hyperpolarized membrane potential especially in conditions where Kir channels are blocked or downregulated. PMID- 16265670 TI - Oligodendrocyte precursor cells generate pituicytes in vivo during neurohypophysis development. AB - In the vertebrate brain, much remains to be understood concerning the origin of glial cell diversity and the potential lineage relationships between the various types of glia. Besides astrocytes and myelin-forming oligodendrocytes, other macroglial cell populations are found in discrete areas of the central nervous system (CNS). They share functional features with astrocytes and oligodendrocytes but also display specific characteristics. Such specialized cells, called pituicytes, are located in the neurohypophysis (NH). Our work focuses on the lineage of the pituicytes during rodent development. First, we show that cells identified with a combination of oligodendrocyte precursor cell (OPC) markers are present in the developing rat NH. In culture, neonatal NH progenitors also share major functional characteristics with OPCs, being both migratory and bipotential, i.e. able to give rise to type 2 astrocytes and oligodendrocytes. We then observe that, either in vitro or after transplantation into myelin-deficient Shiverer brain, pieces of NH generate myelinating oligodendrocytes, confirming the oligodendrogenic potentiality of NH cells. However, no mature oligodendrocyte can be found in the NH. This led us to hypothesize that the OPCs present in the developing NH might be generating other glial cells, especially the pituicytes. Consistent with this hypothesis, the OPCs appear during NH development before pituicytes differentiate. Finally, we establish a lineage relationship between olig1+ cells, most likely OPCs, and the pituicytes by fate-mapping experiments using genetically engineered mice. This constitutes the first demonstration that OPCs generate glial cells other than oligodendrocytes in vivo. PMID- 16265671 TI - Lipopolysaccharide-activated SHP-1-deficient motheaten microglia release increased nitric oxide, TNF-alpha, and IL-1beta. AB - Accumulating evidence suggests a deleterious role for activated microglia in facilitating neuronal death by producing neurocytotoxic substances during injury, infection, or neurodegenerative diseases. After cochlear ablation, abnormal microglial activation accompanied by increased neuronal loss within the auditory brainstem occurs in motheaten (me/me) mice deficient in the protein tyrosine phosphatase SHP-1. To determine whether abnormally activated microglia contribute to neuronal death in me/me mice, primary microglial cultures from me/me and wild type mouse cortices were stimulated by the bacterial endotoxin lipopolysaccharide (LPS) to evaluate the secretion of the neurotoxic mediators nitric oxide (NO), tumor necrosis factor-alpha (TNF-alpha), and interleukin-1beta (IL-1beta). Me/me microglia release significantly greater amounts of all three mediators compared with wild-type microglia. However, the increased release of these compounds in microglia lacking SHP-1 does not appear to occur through activation of extracellular signal-regulated kinase (ERK), p38 kinase subgroups of mitogen activated protein (MAP) kinases, or increases in NF-kappaB-inducing kinase (NIK). These results suggest that abnormal microglial activation and release of neurotoxic compounds may potentiate neuronal death in deafferented cells and can thus potentiate neurodegeneration in the me/me brainstem. Our data also indicate that SHP-1 is engaged in signaling pathways in LPS-activated microglia, but not through regulation of the ERK and p38 MAP kinases. PMID- 16265672 TI - Role of intracellular S100A4 for migration of rat astrocytes. AB - S100A4 is a member of the EF-hand family of calcium-binding proteins, first identified in tumor cells, and implicated in tumor invasion and metastasis. Intracellular upregulation of S100A4 is associated with increased motility of tumor cells. Extracellular application of S100A4 increases the motility of glioma cells in vitro. We showed previously that astrocytes in spinal cord and brain white matter also express S100A4. This expression is markedly increased in reactive white matter astrocytes after injury. Here, we have explored how changes in intracellular S100A4 affect migration of astrocytes. We produced cultures of white matter, S100A4 expressing astrocytes, and developed a small interfering (si) RNA approach to specifically eliminate S100A4 expression in these cells, and compared the migration of astrocytes expressing S100A4 with astrocytes transfected with S100A4 siRNA. As a "positive control" we used S100A4 expressing C6 glioma cells. In contrast to malignant cells, S100A4 expressing astrocytes increased their migration capacity after S100A4 siRNA treatment. At the same time, and in parallel with increased migration, white matter astrocytes increased their expression of metalloproteinases MMP-9 and MT1-MMP. The addition of MMP 2/MMP-9 inhibitor resulted in a significant inhibition of migration in S100A4 siRNA-treated astrocytes. These findings indicate that S100A4 has a stabilizing function in reactive white matter astrocytes, a function that may contribute to the development of a rigid, growth-inhibitory glial scar. PMID- 16265673 TI - Functional expression of NOD2, a novel pattern recognition receptor for bacterial motifs, in primary murine astrocytes. AB - There is growing appreciation that resident brain cells can initiate and/or regulate inflammation after trauma or infection in the central nervous system (CNS). Recent studies from our laboratory have begun to shed light on the mechanisms by which astrocytes perceive bacterial challenges by demonstrating the functional expression of Toll-like receptors (TLR) in this cell type. In the present study, we demonstrate that astrocytes also express members of the novel nucleotide-binding oligomerization domain (NOD) family of proteins that can serve as cytosolic pattern recognition receptors. We show that isolated cultures of murine astrocytes constitutively express robust levels of NOD2, a molecule that can recognize a minimal peptidoglycan motif. Expression of NOD2 is significantly upregulated after exposure to two disparate and clinically relevant bacterial pathogens of the CNS, Borrelia burgdorferi and Neisseria meningitidis. Similarly, NOD2 protein expression is elevated after exposure to specific bacterial ligands for TLRs. Importantly, we show that astrocytes express Rip2 kinase, an essential downstream effector molecule for NOD-mediated cell responses, and demonstrate that this expression is upregulated after bacterial challenge. Furthermore, we confirm the functional nature of NOD2 in astrocytes by demonstrating that a specific ligand for this receptor induces significant inflammatory cytokine production and augments immune responses induced by TLR ligation. Taken together, the present demonstration that astrocytes express functional NOD2 proteins may represent a potentially important mechanism by which this glial cell type initiates either protective host responses within the brain or the progression of damaging CNS inflammation. PMID- 16265674 TI - Topotecan as a radiosensitizer in the treatment of children with malignant diffuse brainstem gliomas: results of a French Society of Paediatric Oncology Phase II Study. AB - BACKGROUND: The current Phase II study was conducted to evaluate the survival and toxicity observed in children with newly diagnosed brainstem gliomas who were treated with the daily radiotherapy with topotecan used as a radiosensitizer. METHODS: Eligible patients were those ages 3-18 years with previously untreated tumors arising in the pons diagnosed within the previous 6 months. Histologic confirmation was not mandatory provided that the clinical and magnetic resonance imaging findings were typical for a diffusely infiltrating brainstem lesion. Treatment was comprised of a 6-week course of topotecan administered intravenously at a dose of 0.4 mg/m(2)/day over 30 minutes within 1 hour before irradiation. Radiotherapy was comprised of a once-daily treatment of 1.8 grays (Gy) per fraction to a total dose of 54 Gy. RESULTS: Thirty-two patients were included in the current study between August 2000 and October 2002. All patients completed the combined treatment in accordance with the treatment design. Only partial responses were observed, occurring in 40% of the patients. The 9-month and 12-month survival rates were 34.4% +/- 8% and 25.5% +/- 8%, respectively. The median duration of survival for these 32 patients was 8.3 months. An intratumoral cystic/necrotic change was observed in five patients, with clinical impairment noted in two patients. One intratumoral hemorrhage occurred during radiotherapy, and was associated with transitory neurologic impairment. CONCLUSIONS: The findings of the current study regarding newly diagnosed brainstem glioma patients treated with topotecan given as a radiosensitizing agent did not reproduce the encouraging results obtained in preclinical studies. Therefore, the concomitant combination of topotecan and radiotherapy at this schedule and these doses cannot be recommended for the treatment of patients with brainstem gliomas. PMID- 16265676 TI - Application of cross-linked beta-cyclodextrin polymer for adsorption of aromatic amino acids. AB - Beta-cyclodextrin (beta-CyD) was cross-linked by hexamethylene diisocyanate and the polymer was investigated for adsorption of aromatic amino acids (AAA) from phosphate buffer. High adsorption rates were observed at the beginning and the adsorption equilibrium was then gradually achieved in about 45 min. The adsorption of AAA decreased with the increase of initial concentration and also temperature. Under the same conditions, the adsorption efficiencies of AAA were in the order of L-tryptophan (L-Trp) > L-phenylalanine (L-Phe) > L-tyrosine (L Tyr). Much higher adsorption values, up to 52.4 and 43.0 mg/g for L-Trp and L Phe, respectively, at 50 mmol/L and 3.2 mg/g for L-Tyr at 2 mmol/L, were obtained with the beta-CyD polymer at 37 degrees C. It was shown that the adsorption of AAA on the beta-CyD polymer was consistent with the Freundlich isotherm equation. The adsorption of mixed aromatic amino acids and branched-chain amino acids (BCAA) showed that AAA were preferentially adsorbed with adsorption efficiencies 10-24%, while those of BCAA were lower than 2%. It seems that the structure and hydrophobicity of amino acid molecules are responsible for the difference in adsorption, by influencing the strength of interactions between amino acid molecule and the polymer. PMID- 16265677 TI - Adding nails to the coffin of underpowered trials. PMID- 16265675 TI - 2C4, a monoclonal antibody against HER2, disrupts the HER kinase signaling pathway and inhibits ovarian carcinoma cell growth. AB - BACKGROUND: Human epidermal growth factor receptor 2 (HER2) is overexpressed in 25-30% of ovarian carcinoma cases and a correlation between increased HER2 expression and decreased survival has been demonstrated. HER2 is a ligand-less member of the HER family that functions as the preferred coreceptor for epidermal growth factor receptor (EGFR), HER3, and HER4. METHODS: An approach was developed to target HER2's role as a coreceptor using a monoclonal antibody, 2C4, which sterically hinders HER2's recruitment into a functional HER complex. RESULTS: HER2 was robustly expressed in all eight ovarian carcinoma cell lines; expression of EGFR and HER3 was variable. Even though four of the eight cell lines responded to EGF, 2C4 antibody moderately inhibited in vitro proliferation of only two cell lines (OVCA433 and SK-OV-3). Furthermore, ligand-stimulated p-MAPK expression was inhibited by 2C4 only in these two cell lines after exposure to EGF. Immunoprecipitation and eTag analysis revealed that OVCA433 expressed heterodimers of EGFR/HER2, and these heterodimers were disrupted after treatment with 2C4, whereas OVCA432 cells did not have these heterodimers. In murine xenograft experiments, the in vivo growth of OVCA433, but not of OVCA432, ovarian carcinoma cells was significantly inhibited by 2C4 treatment of the mice. CONCLUSION: 2C4 is able to disrupt the HER signaling pathway and inhibit the in vitro and in vivo growth of ovarian carcinoma cell lines. The response appears limited to lines in which HER2 heterodimers were able to transduce proliferative signals. Our findings suggest a strong rationale to conduct clinical trials of 2C4 in a subset of patients with ovarian tumors. PMID- 16265678 TI - Discovering that B cells are important in rheumatoid arthritis. PMID- 16265679 TI - Can magnetic resonance imaging of the knee predict future malalignment? PMID- 16265680 TI - Bones, blood vessels, and the immune system: what's the link? PMID- 16265681 TI - Predictive value of antineutrophil cytoplasmic antibodies in small-vessel vasculitis: is the glass half full or half empty? PMID- 16265682 TI - Treatment of hepatitis C virus-related systemic vasculitis. PMID- 16265683 TI - The prevalence of underpowered randomized clinical trials in rheumatology. AB - OBJECTIVE: The conduct of underpowered randomized controlled trials (RCT) has recently been criticized in medical journals. We investigated the current prevalence of underpowered RCT in rheumatology. METHODS: We searched to identify randomized, prospective RCT assessing clinical efficacy of treatments for adult rheumatic diseases published in English in 2001 and 2002. RCT were assessed as positive or negative based on the result of the primary outcome measure. For phase III RCT with negative results without power analysis, we calculated adequate sample size using beta = 0.20 and alpha = 0.05. We also examined trial quality by assessing the adequacy of reported random sequence generation, allocation concealment, and analysis, and compared the quality of reporting of RCT with adequate and inadequate sample size. RESULTS: A total of 228 RCT met inclusion criteria; of the 205 phase III trials, 119 were positive, 81 were negative. The remaining 5 trials made no statistical comparison between interventions, and did not supply enough information for a result to be calculated. Of the 86 negative or indeterminate RCT, 37 reported sample size calculations (all but 4 had adequate power). Of the 49 remaining phase III trials that did not report power calculations, we conducted sample size calculations; only 10 were adequately powered. Few of the underpowered RCT studied rare rheumatic diseases. Negative RCT with inadequate sample size were less likely to describe adequate random sequence generation or allocation concealment than positive RCT or negative RCT with adequate sample size. CONCLUSION: The conduct of underpowered trials is not an infrequent occurrence in rheumatology, with only 50% of negative or indeterminate phase III rheumatology RCT in 2001-2002 having adequate sample size. PMID- 16265684 TI - The predictive value of rheumatoid factor isotypes, anti-cyclic citrullinated peptide antibodies, and antineutrophil cytoplasmic antibodies for mortality in patients with rheumatoid arthritis. AB - OBJECTIVE: To evaluate the significance of rheumatoid factor (RF) and its isotypes (IgA RF, IgG RF, and IgM RF), anti-cyclic citrullinated peptide antibodies (anti-CCP), and antineutrophil cytoplasmic antibodies (ANCA) in predicting mortality in patients with rheumatoid arthritis (RA). METHODS: The study population comprised 604 patients with RA participating in a cross sectional study in 1987. Presence of RF (n = 604), RF isotypes (n = 206), anti CCP (n = 184), and ANCA (n = 200) were determined in these patients from available baseline sera. Vital status was assessed in 1999 and multivariate Cox regression analysis used to compare mortality in RA patients with or without different antibodies. RESULTS: Of the 604 patients with RA, 55% were positive for RF, 66% for anti-CCP, and 14.5% for perinuclear ANCA. Twelve patients (19%) with RF were anti-CCP-negative and 34 (40%) without RF were anti-CCP-positive. Of the total 604 patients, 160 had died by 1999. Positive RF and high IgA and IgM RF levels predicted increased mortality, while positive anti-CCP or ANCA did not. However, high anti-CCP levels were related to an increased mortality risk. CONCLUSION: Patients with RA with positive RF, especially IgA and IgM isotypes, carry a risk of dying earlier than patients without these serological findings. PMID- 16265685 TI - Effects of Angiotensin-converting enzyme inhibition and statin treatment on inflammatory markers and endothelial functions in patients with longterm rheumatoid arthritis. AB - OBJECTIVE: To investigate the effects of angiotensin-converting enzyme (ACE) inhibitors and statins (hydroxy-methyl-glutaryl-CoA reductase inhibitors) on inflammatory markers and endothelial functions in patients with rheumatoid arthritis (RA). METHODS: A total of 45 patients with longterm RA were randomized into 3 groups to receive 8 weeks of treatment with placebo (n = 15), simvastatin (20 mg/day, n = 15), or quinapril (10 mg/day, n = 15) as an adjunct to existing antirheumatic drug treatment. Factors with a role in the development of endothelial dysfunction, such as C-reactive protein (CRP), fibrinogen, nitric oxide (NO), and serum cytokine concentrations including interleukin 1beta (IL 1beta), IL-6, and tumor necrosis factor-alpha (TNF-alpha) were measured at baseline and in the posttreatment period. Brachial artery vasodilator responses were assessed by high resolution ultrasound to evaluate endothelial functions. RESULTS: Simvastatin treatment significantly decreased serum CRP and TNF-a [from 14 +/- 6 to 7 +/- 3 mg/l (p = 0.025) and 30 +/- 5 to 16 +/- 4 pg/ml (p = 0.012), respectively], while quinapril had no significant changes in these 2 measures. IL 1beta and IL-6 showed insignificant changes in patients in the 2 drug groups. Endothelium-dependent vasodilatation was improved significantly in the simvastatin group [from 5.3 +/- 1.1% to 8.9 +/- 1.4% (p = 0.025)], while there was no difference in endothelium-independent vasodilatation [9.0 +/- 1.8% to 11.2 +/- 2.5% (p = 0.17)]. The quinapril group showed no significant changes in both types of vasodilation although there was a tendency to an increase in endothelium dependent vasodilatation [from 6.1 +/- 0.8% to 7.8 +/- 0.7% (p = 0.06)]. Treatment with the 2 drugs had no significant effects on resting arterial diameter. CONCLUSION: We show that simvastatin 20 mg daily improves endothelial function in patients with RA. Its beneficial effect may be attributed to lowering CRP and TNF-alpha concentrations. ACE inhibition with daily 10 mg quinapril was found to have no significant effects on inflammatory markers and endothelial vasodilator response. PMID- 16265686 TI - Modification of pro- and antiinflammatory cytokines and vascular-related molecules by tumor necrosis factor-a blockade in patients with rheumatoid arthritis. AB - OBJECTIVE: Analysis of serum concentrations and modifications of tumor necrosis factor-a (TNF-a), its soluble receptors (TNFR), interleukin 10 (IL-10), and vascular related molecules [soluble vascular cell adhesion molecule 1 (sVCAM-1), vascular endothelial growth factor (VEGF)] after therapy with methotrexate (MTX) and anti-TNF (infliximab) in patients with rheumatoid arthritis (RA). METHODS: Thirty-six patients with RA and 20 healthy controls were included. Patients had been orally taking a stable dose of MTX of at least 12.5 mg/week for a minimum of 6 months before inclusion in the study. Twenty-five patients had shown a clinical response to MTX (MTX Group). The other 11 had shown an unsatisfactory response and presented with active RA; they were selected for additional treatment with infliximab (MTX + IFM Group). Disease activity score (DAS28), hemoglobin concentration, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), and serum levels of TNF-a, soluble TNFR, IL-10, sVCAM-1 and VEGF were determined at baseline and prior to every infusion of infliximab (3 mg/kg) at 2, 6, 14, 22, and 30 weeks. RESULTS: Although serum levels of TNF-a were similar in patients and controls, patients showed significantly higher concentrations of both soluble TNFR (sTNFR55 and sTNFR75), IL-10, sVCAM-1, and VEGF than healthy individuals. Significantly higher levels of sVCAM-1 and VEGF, but not of the other tested molecules, were detected in those with active disease. After infliximab treatment (MTX + IFM Group) there was a significant decrease in DAS28 and modified Health Assessment Questionnaire scores and ESR and CRP levels. Serum concentration of VEGF showed a significant decrease after infliximab, with levels comparable to those of patients with inactive RA, although VEGF continued to present higher values than in healthy controls. CONCLUSION: Increased levels of vascular related molecules sVCAM-1 and VEGF are serum markers of active RA. The absence of normalization of levels of these molecules in patients with inactive RA could be one of the reasons response to therapy is only temporary. PMID- 16265687 TI - Safety and efficacy of rituximab in patients with rheumatoid arthritis refractory to disease modifying antirheumatic drugs and anti-tumor necrosis factor-alpha treatment. AB - OBJECTIVE: To determine the safety and efficacy of rituximab treatment in patients with active seropositive rheumatoid arthritis (RA) who had experienced an inadequate response to treatment with anti-tumor necrosis factor-alpha agents and/or traditional disease modifying antirheumatic drugs. METHODS: Rituximab was administered to 17 patients as weekly infusions for 4 consecutive weeks. Patients continued their baseline therapy and were followed for 28 weeks. RESULTS: All patients were evaluable for safety, and 13 for efficacy. Profound B cell depletion occurred by 12 weeks and was sustained at 24 weeks, whereas T cell, complement, and immunoglobulin levels remained within normal ranges. Rituximab was well tolerated, with no infusion related reactions and only mild/moderate adverse events. American College of Rheumatology 20% response (ACR20) was achieved in 55% of patients by Week 5, 75% by Week 8, 50% at Week 16, and 67% at Week 28. Corresponding ACR50 and ACR70 responses were achieved in 36% and 18%, 25% and 17%, 42% and 25%, and 33% and 17% of patients at Weeks 5, 8, 16, and 28, respectively. There were significant improvements over baseline in tender and swollen joint counts (p < 0.0001), physician's global assessment of disease activity (p = 0.0001), and patient assessed pain (p = 0.0005) and disability (p = 0.0386). Erythrocyte sedimentation rate (p = 0.0361) and rheumatoid factor titers (p < 0.0001) also decreased significantly. CONCLUSION: These results support the hypothesis that B cells play an important role in RA pathophysiology, and suggest that rituximab is effective and well tolerated, with a rapid onset of clinical benefit, in patients with refractory, seropositive active RA. PMID- 16265688 TI - Anti-tumor necrosis factor-alpha therapy augments dipeptidyl peptidase IV activity and decreases autoantibodies to GRP78/BIP and phosphoglucose isomerase in patients with rheumatoid arthritis. AB - OBJECTIVE: To assess the enzymatic activity and biochemical status of dipeptidyl peptidase IV (DPP IV), an enzyme that participates in the degradation of proinflammatory molecules, in sera from a group of patients with rheumatoid arthritis (RA; n = 15) treated with a human anti-tumor necrosis factor-a (anti TNF-alpha) antibody (adalimumab) for 32 weeks. IgG antibody titers against chaperone Bip (GRP78), phosphoglucose isomerase (PGI), lactate dehydrogenase (LDH), fibronectin (FN), and actin were also studied. METHODS: DPP IV activity was measured in sera using Gly-Pro-p-nitroanilide as substrate. The biochemical profile of circulating DPP IV glycoforms was assessed by isoelectric focusing gel electrophoresis. All IgG autoantibody titers and their sialylation levels were determined by ELISA. RESULTS: Patients showed significant increases in serum DPP IV enzymatic activity from basal values (3.554 +/- 1.096) with respect to those obtained at 32 weeks (4.787 +/- 0.953; p < 0.05). Changes in the biochemical profile of circulating DPP IV from acidic to more neutral isoelectric point glycoforms were also seen during treatment. The elevated titers of anti-GRP78 and anti-PGI IgG observed at the beginning of treatment decreased significantly during therapy, whereas those of anti-LDH, anti-FN, and anti-actin IgG remained unchanged. At the end of treatment, sialylation levels of anti-GRP78 and anti-PGI IgG antibodies increased to nearly normal levels. The DPP IV biochemical changes were accompanied by a significant improvement of the Disease Activity Score (DAS28). CONCLUSION: The reduced activity of DPP IV along with increased titers of circulating antibodies to GRP78 and PGI may play a role in the pathogenesis of RA and can be successfully modified by administration of adalimumab. PMID- 16265690 TI - Skin cancer, rheumatoid arthritis, and tumor necrosis factor inhibitors. AB - OBJECTIVE: To determine the rates of reported non-melanoma skin cancer (NMSC) in a large cohort of patients with rheumatoid arthritis (RA) in comparison to patients with osteoarthritis (OA) and to determine risk factors for the development of NMSC in patients with RA. METHODS: Self-reported information from 15,789 patients with RA and 3,639 patients with OA were collected through semi annual questionnaires since 1999. Survival analyses were used to determine incidence rates for NMSC among patients with RA and OA. Multivariate Cox proportional hazard models were used to estimate hazard ratios (HR) for the development of NMSC. Separate analyses were performed for patients with RA to explore associations between use of immunosuppressive medication and development of NMSC. RESULTS: The crude (unadjusted) incidence rate for reported NMSC among patients with RA and OA were 18.1 and 20.4 per 1000 patient years, respectively. OA patients were older, more likely to be Caucasian, and had higher past incidence of NMSC. Age, male sex, Caucasian race, and history of NMSC prior to entry into the database were associated with an increased risk of NMSC in multivariate Cox proportional hazard models. After adjustment for covariates, RA was associated with an increased risk of NMSC (HR 1.19, p = 0.042). Among RA patients, the development of NMSC was associated with use of prednisone (HR 1.28, p = 0.014) and tumor necrosis factor (TNF) inhibitors alone or with concomitant methotrexate (HR 1.24, p = 0.89 and HR 1.97, p = 0.001, respectively) in addition to established risk factors including fair skin, age, male sex, and previous history of NMSC. No association was found between use of methotrexate or leflunomide and development of NMSC (HR 1.12, p = 0.471, HR 0.83, p = 0.173, respectively). CONCLUSION: In this large, national cohort, RA was associated with an increased risk for development of NMSC. Among patients with RA, use of TNF inhibitors and prednisone were associated with an increased risk of NMSC. PMID- 16265689 TI - Staphylococcus aureus in patients with rheumatoid arthritis under conventional and anti-tumor necrosis factor-alpha treatment. AB - OBJECTIVE: To compare the prevalence of nasal and oral Staphylococcus aureus in patients with rheumatoid arthritis (RA) with the prevalence in controls with other rheumatic diseases, and to determine predictors of S. aureus carriage and the influence of treatment with anti-tumor necrosis factor-a (anti-TNF-alpha) agents. METHODS: Eighty-one patients with RA and 83 other control patients of 2 outpatient rheumatology clinics were cultured for nasal and oral carriage of S. aureus. Quantitative nasal cultures for S. aureus were performed from swabs of the anterior nares, the posterior pharynx, and the soft palate. Information on medications, medical conditions, and risk factors for S. aureus carriage was collected from all participants by a questionnaire and confirmed by chart review. RESULTS: The S. aureus carriage rate (nasal and/or oral colonization) was 34.6% among RA patients and 32.5% among controls (p = 0.87). Being treated with an anti TNF-alpha agent plus methotrexate (MTX) was the only independent predictor of S. aureus carriage (OR 3.24, 95% CI 1.16-9.05, p = 0.025). The S. aureus carriage rate among RA patients treated with an anti-TNF-alpha agent plus MTX was 60% (9/15) versus 23.1% (3/13) in RA patients treated with an anti-TNF-alpha agent only (p = 0.049). All S. aureus isolates were susceptible to oxacillin. CONCLUSION: The S. aureus carriage rate among patients with RA was not higher than among controls. Treatment with anti-TNF-alpha agents was not associated with an increased S. aureus carriage rate. However, treatment with an anti-TNF-alpha agent plus MTX may predispose patients to S. aureus carriage. PMID- 16265691 TI - Static wrist splint use in the performance of daily activities by individuals with rheumatoid arthritis. AB - OBJECTIVE: In individuals with rheumatoid arthritis (RA), to identify the influence of wrist splint wear on pain, work performance, endurance, perceived task difficulty, and perceived splint benefit while performing various upper limb tasks. METHODS: This crossover study included 30 individuals with wrist involvement. Pain, work performance, endurance, and perceived task difficulty were assessed with the splint on and off. Using a work simulator, participants performed 14 tasks, 10 assessing work performance and 4 assessing endurance. A visual analog scale (VAS) was used to rate pain, task difficulty, and perceived splint benefit. RESULTS: With the splint on, pain was significantly lower in 5 tasks, as was perceived difficulty in task performance. Work performance did not differ significantly with the splint on versus off. While mean endurance scores were always better with the splint on, differences reached significance on only one task. The task with greatest overall perceived splint benefit was "chopping with a knife." CONCLUSION: Results revealed that for most tasks, there was generally a positive effect of splint use on hand function; however, perceived splint benefit was marginal. For most tasks splint use improved or did not change pain levels, did not interfere with work performance, increased or maintained endurance, and did not increase perceived task difficulty. The findings suggest that wrist splint prescription is not a simple process; clinicians and clients need to work together to determine the daily wear pattern that maximizes benefit and minimizes inconvenience according to the client's individual needs. PMID- 16265692 TI - The clinical spectrum of antinuclear antibodies associated with the nuclear dense fine speckled immunofluorescence pattern. AB - OBJECTIVE: Autoantibodies to lens epithelium-derived growth factor (LEDGF) depict a distinctive nuclear dense fine speckled (DFS) pattern in the indirect immunofluorescence antinuclear antibody assay (IIF-ANA). Definition of the clinical spectrum associated with anti-LEDGF antibodies has been evolving over the last decade. We investigated the frequency, clinical spectrum, and immunologic specificity of the DFS pattern in a general clinical laboratory routine. METHODS. All serum samples entered for IIF-ANA determination within a 2 year period were examined for the DFS pattern. Positive samples with consistent clinical information were studied further by IIF with isotype-specific conjugate and immunoblot analysis. RESULTS: Among 13,641 ANA-positive samples, 5081 (37%) presented the DFS pattern. Within a 6 month nested period, there were 650 samples with DFS pattern, and consistent clinical data were available for 81 of these. DFS reactivity was mainly due to IgG. Most samples (86%) presented titer > or = 1/640. Eighty of the 81 DFS samples reacted with a 75 kDa band that comigrated with the band elicited by the standard anti-LEDGF serum. Antibodies that were affinity-purified from the 75 kDa band reproduced the DFS pattern on IIF-ANA. The clinical spectrum associated with DFS reactivity included autoimmune diseases (39%) and an array of nonautoimmune conditions (61%). Among the autoimmune patients, over half presented evidence of autoimmune thyroiditis. CONCLUSION: Anti-LEDGF/p75 antibodies are a common finding among ANA-positive individuals with no evidence of rheumatic autoimmune disease, and should be regarded as a low specificity finding even when in moderate or high titer. PMID- 16265693 TI - Systemic lupus erythematosus in a multiethnic cohort (LUMINA): XXIX. Elevation of erythrocyte sedimentation rate is associated with disease activity and damage accrual. AB - OBJECTIVE: To determine if different categories of erythrocyte sedimentation rate (ESR) elevation are associated with disease activity and/or damage in systemic lupus erythematosus (SLE). METHODS: We studied 2317 study visits in 553 SLE patients (> or = 4 American College of Rheumatology criteria, < or = 5 years' disease duration at enrollment) from a multiethnic (Hispanic, African American, and Caucasian) longitudinal study of outcome. A study visit was done every 6 months for the first year and annually thereafter. Erythrocyte sedimentation rate (ESR) was measured using the Westergren method; results were expressed in 4 categories: < 25 (normal), 25-50 (mild elevation), 51-75 (moderate elevation), and > 75 (marked elevation) mm/h. Anti-dsDNA antibodies were measured at enrollment with the Crithidia luciliae assay. Disease activity was assessed with the Systemic Lupus Activity Measure (SLAM) and the Physician's Global Assessment (PGA). Because ESR is one of the measures evaluated in the SLAM, it was excluded from the total SLAM score. Disease damage was assessed with the Systemic Lupus International Collaborating Clinics damage index (SDI). The relationship between the SLAM (total and PGA) and SDI scores (at baseline and for all visits) and anti dsDNA antibodies (at enrollment) with ESR was examined by univariable and generalized estimating equation (GEE) regression analyses. Ethnicity, age, and sex were entered in all regression models. RESULTS: The cohort consisted of 89.7% women with mean age 36.8 (SD 12.6) years and disease duration 4.6 (SD 3.2) years. GEE analyses showed that increasing levels of ESR and anti-dsDNA antibody positivity were independently associated with SLAM and PGA scores, at enrollment and for all visits. Overall, the associations of ESR with SLAM and PGA scores were stronger than for the presence of anti-dsDNA antibodies. At baseline, there was no relationship of ESR elevation or anti-dsDNA positivity with SDI scores. However, when all visits were studied, moderate and marked elevations of ESR were independently associated with SDI scores. CONCLUSION: Mild, moderate, and marked ESR elevations are strongly associated with disease activity in SLE. Moderate and marked ESR elevations are also associated with damage accrual. These associations are stronger than those for the presence of anti-dsDNA antibodies. Our data suggest that ESR could be used to assess disease activity and predict organ/system damage in a relatively rapid and inexpensive manner in SLE. PMID- 16265694 TI - Enhanced expression of programmed death-1 (PD-1)/PD-L1 in salivary glands of patients with Sjogren's syndrome. AB - OBJECTIVE: Programmed death-1 (PD-1) mediates a negative signal and introduces tolerance for lymphocytes. Dysfunction of the PD-1 pathway is thought to result in autoimmune diseases such as rheumatoid arthritis (RA). To investigate the role of the PD-1/PD-L system in the pathology of Sjogren's syndrome (SS), we examined the expression of PD-1 and its ligand PD-L1 in salivary lymphocytes and salivary glands from patients with SS. METHODS: Flow cytometry analysis was used to determine expression of PD-1 in SS salivary lymphocytes. Intracellular staining of interleukin 10 (IL-10) was performed after stimulation with PMA and ionomycin. Indirect immunohistochemistry was used to investigate the expression of PD-1 and PD-L1. RESULTS: The mean fluorescence intensity of PD-1 expression in SS salivary lymphocytes was significantly higher than that from healthy controls and patients with RA or systemic lupus erythematosus. PD-1-positive SS salivary lymphocytes expressed IL-10 intracellularly upon PMA/ionomycin stimulation. Immunohistochemical analysis showed that PD-1 was expressed on infiltrating lymphocytes in salivary gland from 52% of SS patients, and PD-L1 was expressed on ductal and acinar epithelial cells from 68% of SS patients. In vitro analysis using HSG cells revealed that PD-L1 was induced by interferon-gamma but not by tumor necrosis factor-alpha and IL-1beta. CONCLUSION: PD-1 is expressed on T lymphocytes and PD-L1 on epithelial cells from inflamed salivary glands of patients with SS, which suggests that dysfunction of the PD-1/PD-L1 pathway may be related to tolerance for lymphocytes, which causes SS. PMID- 16265695 TI - Serum relaxin in systemic sclerosis. AB - OBJECTIVE: To evaluate serum levels of relaxin (RLX), a hormone with acknowledged antifibrotic activity, in patients with systemic sclerosis (SSc). METHODS: We performed a pilot study of 50 outpatients with SSc and 50 healthy subjects. Serum RLX was measured using the relaxin ELISA. Statistical analysis was performed using Student's t test. RESULTS: Serum RLX appeared to be significantly higher (p < 0.001) in patients with SSc compared to controls. RLX appeared significantly increased (p < 0.001) in male patients compared to male controls, and in female patients compared to female controls. RLX was significantly higher (p < 0.001) in female patients and female controls compared to male patients and male controls. CONCLUSION: In patients with SSc, the increased level of RLX represents a defensive response against the fibrotic process. PMID- 16265696 TI - Predictive value of antineutrophil cytoplasmic antibodies in small-vessel vasculitis. AB - OBJECTIVE: . The predictive value of antineutrophil cytoplasmic antibodies (ANCA) as markers of clinical activity of small vessel vasculitis is controversial. We reviewed the outcome of patients with ANCA-associated vasculitis from a single center and evaluated the predictive value of ANCA as markers of relapse. METHODS: The medical history of all consecutive patients with ANCA-associated vasculitis followed at our outpatient clinic was retrospectively reviewed. ANCA were monitored by immunofluorescence and by ELISA (antiproteinase 3 and antimyeloperoxidase). RESULTS: Twenty-three patients with Wegener's granulomatosis and 13 with microscopic polyangiitis were followed for a median period of 5 years (1 mo-16 yrs). Most patients were treated with combined corticosteroids and cyclophosphamide. In the Wegener's granulomatosis group, remission was obtained in 21 of 23 patients (91%), within 4-48 weeks (median 7.5); relapses occurred in 74%. In the microscopic polyangiitis group, remission was obtained in all patients within 3-38 weeks (median 8); relapses occurred in 33%. In contrast to persistently (> 6 mo) elevated ANCA titers, which were not significantly associated with disease relapse, the predictive value of an acute rise in ANCA titers was strongly associated with the magnitude of the increase. CONCLUSION: Our study demonstrates the weak predictive value of persistently elevated ANCA titers, in contrast to acute rises in ANCA titers. Although an acute rise in ANCA titers may help in a decision whether to introduce immunosuppressive therapy, the final decision has to be based on both clinical and laboratory markers. PMID- 16265697 TI - Interleukin-6 promoter polymorphism at position -174 in giant cell arteritis. AB - OBJECTIVE: To investigate potential associations between the -174 G/C interleukin 6 (IL-6) promoter polymorphism and susceptibility to and clinical features of giant cell arteritis (GCA), particularly in patients with or without polymyalgia rheumatica (PMR) and with or without ischemic complications. METHODS: One hundred and twenty-six patients with biopsy-proven GCA who were residents in Reggio Emilia, Italy, and 112 population-based controls from the same geographic area were genotyped for IL-6 polymorphism at position -174 by molecular methods. Patients were divided in subgroups according to presence or absence of PMR and ischemic complications (visual loss, jaw claudication, cerebrovascular accidents, aortic arch syndrome). RESULTS: Distribution of the G/C 174 genotype was similar in patients with GCA and controls. No significant associations with the IL-6 promoter polymorphism at position -174 were found when GCA patients with or without PMR or with or without ischemic complications were compared. Further, IL 6 genotypes did not significantly affect levels of C-reactive protein or other inflammatory markers at diagnosis. CONCLUSION: Our findings show that the 174 G/C promoter IL-6 polymorphism does not seem to be implicated in susceptibility to and clinical expression of GCA. PMID- 16265698 TI - Association of a functional inducible nitric oxide synthase promoter variant with susceptibility to biopsy-proven giant cell arteritis. AB - OBJECTIVE: To assess the contribution of 2 polymorphisms within the inducible nitric oxide (NOS2A) promoter region to susceptibility to giant cell arteritis (GCA). METHODS: One hundred three patients with biopsy-proven GCA and 198 ethnically matched controls from the Lugo region (Northwest Spain) were studied. Patients and controls were genotyped using polymerase chain reaction techniques for a multiallelic (CCTTT)n and for the TAAA repeat polymorphism in the promoter region of the NOS2A gene. RESULTS: No significant differences in allele or genotype frequencies for the (CCTTT)n repeat polymorphism in the NOS2A gene between patients with GCA and controls were observed. However, significant differences for the TAAA repeat polymorphism between patients and controls were found. The overall distribution of NOS2A TAAA genotypes in patients with biopsy proven GCA was significantly different than controls (p = 0.026). Patients with GCA had an increased frequency of the NOS2A TAAA+ allele (16.5%) compared with controls (9.1%) (p = 0.007; OR 1.98; 95% CI 1.20-3.27). This was due to an increased frequency of both heterozygotes (27.2%) and homozygotes (2.9%) for NOS2A TAAA+ observed in patients compared to controls (15.2% and 1.5%, respectively) (p = 0.007; OR 2.15; 95% CI 1.23-3.78). CONCLUSION: Our results suggest a potential implication for NOS2A TAAA gene polymorphism in GCA susceptibility. PMID- 16265699 TI - Infliximab to etanercept switch in patients with spondyloarthropathies and psoriatic arthritis: preliminary data. AB - OBJECTIVE: To report early experience of switching anti-tumor necrosis factor-a (TNF-alpha) therapy from infliximab to etanercept in patients with spondyloarthropathy (SpA) and psoriatic arthritis (PsA). METHODS: Thirteen patients with various SpA (7 with ankylosing spondylitis and 6 with undifferentiated SpA) and 2 patients with PsA were receiving infliximab. Because they were experiencing inadequate response or adverse events, therapy was changed to etanercept. Patients were evaluated for response to the change in anti-TNF alpha therapy at baseline, after 3 months, and then every 6 months. RESULTS: During the mean 10-month followup after the change in therapy, 9 of 13 patients with SpA and both patients with PsA responded to etanercept and none experienced intolerance to this agent. CONCLUSION: These data suggest that switching between anti-TNF-alpha drugs may be useful for patients with SpA who are unresponsive or intolerant to a first anti-TNF-alpha agent. PMID- 16265700 TI - Female gout: clinical and laboratory features. AB - OBJECTIVE: To evaluate and compare clinical and laboratory features of gout in men and women. METHODS: Twenty-seven women and 31 men with gout underwent clinical and laboratory evaluation and review of medical records. RESULTS: Disease onset in women was a mean of 7 years later than in men. There were no differences between women and men regarding systemic hypertension, diabetes mellitus, hyperlipidemia, chronic renal failure, renal stones, ischemic heart disease, or heavy alcohol consumption. Tophaceous gout was similar in both groups, although female gender seemed to be protective against risk of developing tophi (odds ratio: 0.449; 95% confidence interval: 0.151-1.330). Podagra was more common in men, and women showed a higher frequency of upper limb joint involvement. Most patients had low urate excretion rates. Achieving disease control was similar in women and men. Of the 8 women who were premenopausal at disease onset, 7 had secondary causes for gout; 5 of the 8 had high serum urate despite treatment. CONCLUSION: Gout in women had a later onset and higher frequency of upper limb joint involvement in comparison to men. Those with premenopausal onset tended to be refractory to standard therapy. PMID- 16265701 TI - Prevalence and risk factors for urolithiasis in primary gout: is a reappraisal needed? AB - OBJECTIVE: To assess the prevalence and risk factors for urolithiasis in primary gout. METHODS: One hundred forty patients with primary gout were studied. Urolithiasis was defined as a history of urolithiasis, or nephrolithiasis detected via ultrasonography in patients with no previous record of urolithiasis. Patient age, duration of gout, presence of tophi, obesity, alcoholism, high blood pressure, diabetes, hyperlipidemia, family history of urolithiasis, daily urine output, uricemia, urine pH, FeNa, FeUrate, urine pH/FeUrate index, and daily urine excretion of urate, sodium, calcium and potassium were compared between lithiasic and non-lithiasic subjects. RESULTS: Fifty-five (39%; 95% CI 31-47) patients had urolithiasis, of which 37 (26%) were diagnosed by clinical history and 18 (13%) by ultrasonography. Patients with a silent kidney stone diagnosed by ultrasound tended to have shorter evolution of gout. Aside from urinary H+ ion concentration (lithiasic subjects 5.17 +/- 3.9 microM/l; non-lithiasic subjects 3.80 +/- 3.01 microM/l; p = 0.02), no difference was found between lithiasic and non-lithiasic subjects for the other variables studied. CONCLUSION: Ultrasonography increased the probability of diagnosing urolithiasis by 50%, meaning the prevalence of urolithiasis in gout is likely higher than previously reported. A higher urinary H+ ion concentration was the only variable associated with urolithiasis. Due to advances in diagnosis of gout and urolithiasis, as well as biochemical assays, the prevalence and risk factors for urolithiasis in gout require reassessment. PMID- 16265702 TI - Structural factors associated with malalignment in knee osteoarthritis: the Boston osteoarthritis knee study. AB - OBJECTIVE: Osteoarthritis (OA) is a multifactorial condition. The progression of knee OA is determined in part by mechanical effects on local structures. One of the mechanical influences on cartilage loss is limb alignment. We explored the structural factors associated with malalignment in subjects with symptomatic OA. METHODS: We conducted a cross-sectional assessment using The Boston Osteoarthritis of the Knee Study, a natural history study of symptomatic knee OA. Baseline assessments included knee magnetic resonance imaging (MRI) and information on weight and height. Long-limb radiographs to assess mechanical alignment were obtained at 15 months. Subarticular bone attrition, meniscal degeneration, anterior and posterior cruciate ligament integrity, medial and lateral collateral ligament integrity, marginal osteophytes, and cartilage morphology were assessed on MRI using a semiquantitative, multi-feature scoring method (Whole-Organ MRI Score) for whole-organ evaluation of the knee that is applicable to conventional MRI techniques. We also quantified the following meniscal position measures on coronal MRI images in both medial and lateral compartments: subluxation, meniscal height, and meniscal covering of the tibial plateau. Using the long-limb radiographs, mechanical alignment was measured in degrees on a continuous scale. The purpose of this cross-sectional analysis was to determine the individual and relative contribution of various structural factors to alignment of the lower extremity. We assessed the cross-sectional association between various structural factors and alignment of the lower extremity using a linear regression model. RESULTS: The 162 subjects with all measures acquired had a mean age of 67.0 years (SD 9.2), body mass index 31.4 (SD 5.6); 30% were female and 77% of knees had a Kellgren-Lawrence grade > or = 2. The main univariate determinants of varus alignment in decreasing order of influence were medial bone attrition, medial meniscal degeneration, medial meniscal subluxation, and medial tibiofemoral cartilage loss. Multivariable analysis revealed that medial bone attrition and medial tibiofemoral cartilage loss explained more of the variance in varus malalignment than other variables. The main univariate determinants of valgus malalignment in decreasing order of influence were lateral tibiofemoral cartilage loss, lateral osteophyte score, and lateral meniscal degeneration. CONCLUSION: Cartilage loss has been thought to be the major determinant of alignment. We found that other factors including meniscal degeneration and position, bone attrition, osteophytes, and ligament damage contribute to the variance of malalignment. Further longitudinal analysis is required to determine cause and effect relationships. This should assist researchers in determining strategies to ameliorate the potent effects of this mechanical disturbance. PMID- 16265703 TI - Relationship between bone markers and knee cartilage volume in healthy men. AB - OBJECTIVE: To determine the relationship between biochemical bone markers and knee cartilage volume and cartilage loss over 2 years; and to investigate whether bone markers are useful to predict the cartilage loss in healthy men. METHODS: Forty healthy Caucasian men (mean age 52.3 yrs) with no symptoms of osteoarthritis (OA) were recruited. Each subject had magnetic resonance imaging (MRI) performed on his dominant knee at baseline and 2 years later. Serum level of osteocalcin (OC), urinary levels of pyridinoline (PYD) and deoxypyridinoline (DPD), and total body bone mineral content (BMC) were measured at baseline. Tibial plateau bone size was measured at baseline. Tibial cartilage volume was measured at baseline and at followup, by means of image processing. RESULTS: Twenty-eight men (70%) completed the longitudinal MRI component of the study. At baseline, no significant associations were observed between values of serum OC or urine PYD and DPD and tibial cartilage volume. Higher baseline serum OC level tended to be associated with a decreased rate of cartilage loss (p = 0.06); no significant association was shown between baseline urine PYD and DPD and tibial cartilage loss, after adjusting for age, body mass index, total body BMC, and tibial plateau bone size. CONCLUSION: Higher baseline serum OC level tended to be associated with a decreased rate of cartilage loss, suggesting that increased bone formation may protect against tibial cartilage loss over 2 years. Studies are needed to determine the role of bone metabolism in the pathogenesis of knee OA. PMID- 16265704 TI - Increased monocyte chemoattractant protein-1 in knee joints of rats with adjuvant induced arthritis: in vivo microdialysis. AB - OBJECTIVE: Imbalance of inflammatory and antiinflammatory cytokines plays a critical role in the pathogenesis of rheumatoid arthritis (RA). Precise determination of these cytokines would lead to better understanding of the progression of RA. METHODS: We developed an in vivo microdialysis technique to directly monitor cytokine profiles in knee joints of rats with adjuvant-induced arthritis (AIA). Microdialysates drained from knee joints of rats with AIA and controls were collected and cytokine concentrations were measured by ELISA. Pathological changes of the knee joints and the source of monocyte chemoattractant factor-1 (MCP-1) secretion were also determined by histology and immunohistochemistry. RESULTS: MCP-1 expression in knee joints was significantly higher in AIA rats with erosive changes in their ankles than in normal rats, while interleukin 6 (IL-6) levels were similar in both cases. IL-1beta and interferon-gamma were not detectable in the microdialysates. Increased synovial proliferation and mononuclear inflammatory infiltrates were observed. Synovial cells and mononuclear inflammatory cells expressed both MCP-1 and its receptor, CCR2. CONCLUSION: Our results indicate that the in vivo microdialysis technique is capable of detecting cytokines in the knee joints of rats. Increased expression of MCP-1 and CCR2 in knee joints of AIA rats suggests a role for this cytokine in triggering the mechanisms involved in the pathogenesis of knee joint after ankle erosion. PMID- 16265705 TI - Risk of hospitalization with peptic ulcer disease or gastrointestinal hemorrhage associated with nabumetone, Arthrotec, diclofenac, and naproxen in a population based cohort study. AB - OBJECTIVE: To identify the unbiased differences in the risk of hospitalization with peptic ulcer disease (PUD) or gastrointestinal (GI) hemorrhage among populations using 4 nonsteroidal antiinflammatory drugs (NSAID): nabumetone, Arthrotec, diclofenac plus a cytoprotective agent dispensed separately (diclo+coRx), and naproxen. METHODS: A population based historical cohort study using linked data from provincial healthcare databases. The population of the province of Saskatchewan, Canada, entitled to drug plan benefits in 1995 was eligible (roughly 91% of 1 million people). Participants were identified if they filled a prescription for one of the 4 study NSAID (18,424 individuals). They were then followed for 6 months to determine outcomes. Logistic regression was used to produce estimates of the risk of admission to hospital with a primary diagnosis of PUD or GI hemorrhage associated with the study drugs unbiased by known confounders. RESULTS: Compared to Arthrotec the adjusted odds of hospitalization for PUD for participants taking nabumetone was 2.6 (95% CI 1.0 6.6), diclo+coRx 6.8 (95% CI 3.5-13.4), and naproxen 7.9 (95% CI 3.9-15.9). Compared to nabumetone the adjusted odds of hospitalization for PUD for participants taking diclo+coRx was 2.7 (95% CI 1.2-6.0) and naproxen 3.1 (95% CI 1.3-7.1). No significant differences were noted in terms of admissions for GI hemorrhage. CONCLUSION: Participants taking nabumetone and Arthrotec had significantly lower risk of hospitalization for PUD than those taking the other study drugs. Arthrotec was superior to nabumetone in a head to head comparison and especially when compared with the diclo+coRx and naproxen groups. No short term differences were seen in the rates of admission for GI hemorrhage. It appears that inherent gastroprotective strategies with Arthrotec and to a lesser extent with nabumetone do translate into decreased serious GI side effects at the population level in the short term. PMID- 16265706 TI - Patterns of use and public perception of over-the-counter pain relievers: focus on nonsteroidal antiinflammatory drugs. AB - OBJECTIVE: To assess the frequency and indications for over-the-counter (OTC) nonsteroidal antiinflammatory drug (NSAID) use and to what degree the public is aware of their side effects. METHODS: Two surveys totaling 9062 respondents were performed of the American public. The Roper survey, conducted in 1997, and the National Consumers League (NCL) survey, conducted in December 2002, were intended primarily to assess the public's use of and attitudes toward NSAID and OTC analgesics. RESULTS: Ibuprofen based drugs were the most frequently used OTC in both surveys (57% Roper, 33% NCL). In the Roper survey, 17% of respondents used NSAID, with 38% using both prescription and OTC. Forty-six percent of exclusive OTC users believed OTC were safer, while 56% of exclusive users of prescription NSAID believed they were safer. Sixty percent and 29% of exclusive OTC users were neither aware of nor believed they were at risk for side effects from NSAID, respectively. Twenty-six percent of respondents used more than the recommended dose on the label, while 22% believed warning symptoms would always precede any NSAID induced complications. In the NCL survey, 83% had used an OTC agent in the last year, with 15% reporting daily use, and 49% were not concerned about potential side effects. In this survey, 30% believed there was less risk with OTC analgesics, and 44% consumed more than the recommended dosage on the label. CONCLUSION: OTC analgesics including NSAID are widely used, are frequently taken inappropriately and potentially dangerously, and users are generally unaware of the potential for adverse side effects. Educational intervention directed toward both patients and physicians appears warranted. PMID- 16265707 TI - Primary Sjogren's syndrome in children and adolescents: are proposed diagnostic criteria applicable? AB - OBJECTIVE: To compare the proposed criteria for the diagnosis of primary Sjogren's syndrome (pSS) in childhood to the validated American-European Consensus Group (AECG) classification criteria for pSS in adults. METHODS: Charts of 7 children with pSS seen at British Columbia's Children's Hospital (BCCH) and data on 128 children identified through Medline in the English language literature between 1963 and 2003 were reviewed for pediatric and AECG criteria for pSS. The presence of > or = 4 criteria was required to satisfy the respective classification criteria. The expert clinical opinion of pediatric rheumatologists was considered the gold standard for diagnosis. RESULTS: A total of 24/62 (39%) cases satisfied the AECG criteria; 47/62 (76%) satisfied the proposed pediatric criteria. Inclusion of recurrent parotitis increased the sensitivity of the pediatric clinical criteria. From the cases, 78/133 (59%) satisfied the pediatric oral symptom criteria; only 6/78 (8%) had xerostomia in the absence of recurrent parotitis. There was no reported case of recurrent conjunctivitis in the absence of keratoconjunctivitis sicca. We found 101/130 (78%) cases had at least one positive autoantibody test result [antinuclear antibodies (ANA), rheumatoid factor (RF), SSA, SSB]; 78/123 (63%) had autoantibodies to SSA or SSB. CONCLUSION: The AECG adult criteria for pSS should not be applied to children as the sensitivity is unacceptably low. The inclusion of recurrent parotitis increases the sensitivity of the pediatric criteria, and recurrent parotitis should alert the clinician to the possibility of pSS. The inclusion of recurrent conjunctivitis did not improve the sensitivity over the AECG ocular criteria. The addition of ANA and RF to the AECG criteria did not change the number of patients satisfying the criteria for pediatric pSS. PMID- 16265708 TI - Osteoprotegerin serum levels in Kawasaki disease: an additional potential marker in predicting children with coronary artery involvement. AB - OBJECTIVE: Emerging evidence from in vitro studies and mouse genetics attributes to osteoprotegerin (OPG), a member of the tumor necrosis factor receptor family, an important role in vascular biology. We evaluated serum levels of OPG in a group of children with Kawasaki disease (KD), before immunoglobulin (IVIG) infusion and at 3-month followup. METHODS: Fifty patients (38 boys, 20 girls, median age 3.6 yrs, range 4 mo-7.4 yrs) fulfilling criteria for the diagnosis of KD, 30 febrile controls with infectious diseases, 18 patients with juvenile systemic lupus erythematosus (JSLE), and 40 healthy controls were enrolled. All KD patients received IVIG treatment within the first 10 days of illness, and aspirin. Coronary artery abnormalities (CAA) were reported in 6 out of 58 patients; all were male and younger than 5 years. Serum OPG was measured by ELISA in patients with KD before IVIG and at 3-month followup (median time 3.2 mo, range 3-3.5). RESULTS: At baseline and at the 3-month followup, KD patients had significantly higher OPG serum levels than febrile controls (p < 0.001 and p < 0.004, respectively), JSLE patients (p < 0.0001), and healthy controls (p < 0.0001). At baseline, KD patients who developed CAA had higher OPG serum levels than those without CAA (p = 0.0001); this difference was not present at 3-month followup. The optimal OPG cutoff value of 123.2 pg/ml was a significant predictor for CAA, with a sensitivity of 100% (6/6), a specificity of 96% (50/52), and a positive predictive value of 75% (6/8). CONCLUSION: High OPG levels might be the result of compensatory production during acute and subacute phases of KD. OPG assay might be an additional clinically useful marker to monitor and differentiate patients who develop, from those who do not develop, such coronary artery abnormalities. PMID- 16265709 TI - Magnetic resonance imaging in pachydermodactyly. AB - The magnetic resonance imaging (MRI) findings of a teenage boy with pachydermodactyly are described. The findings include fusiform soft tissue swelling around the proximal interphalangeal joints of the hands, with sparing of the tendons and ligaments. There was no capsular involvement and no evidence of periosteal reaction. PMID- 16265710 TI - Lymphomatoid granulomatosis: a rare mimicker of vasculitis. AB - Lymphomatoid granulomatosis (LG) is a rare Epstein-Barr virus-associated lymphoproliferative disorder, with a propensity for blood vessel destruction. Although it most commonly affects the lung, it can affect multiple extrapulmonary sites (i.e., skin, nervous system, gastrointestinal tract, liver, spleen, kidney, and heart). Since LG often mimics systemic vasculitis, it presents a diagnostic clinical challenge. We describe a case of LG with no pulmonary involvement, which was thought to be systemic vasculitis on the basis of multiorgan involvement and biopsy findings. PMID- 16265711 TI - Progress since OMERACT 6 on including patient perspective in rheumatoid arthritis outcome assessment. AB - The first OMERACT Patient Perspective Workshop took place at OMERACT 6 in 2002. Through a series of meetings and discussion sessions a research agenda emerged and this report outlines progress made on this agenda. Work on identifying novel outcomes, instruments, and methods has shown similarities across European countries in the importance patients with rheumatoid arthritis (RA) attach to specific outcomes, in particular fatigue. Validation of an appropriate instrument to measure fatigue in patients with RA is currently being investigated. Frequent or repeated real-time assessment of symptoms such as pain and fatigue is becoming possible using electronic systems. An OMERACT Patient Panel has been established, and has produced a glossary for patients involved in supporting clinical research. In some centers, efforts are being made to provide Patient Research Partners with knowledge and skills that will enhance their contribution, and some of these approaches will be incorporated into OMERACT 7. The research agenda that was developed during the first Patient Perspective Workshop has stimulated new work in several areas. In addition, international attention has been drawn to the need to make sure that the patient's perspective is not lost among the technical expertise of rheumatology. PMID- 16265712 TI - Incorporating the patient perspective into outcome assessment in rheumatoid arthritis--progress at OMERACT 7. AB - The Patient Perspective Workshop at OMERACT 7 addressed the question of assessing the outcomes of intervention in rheumatoid arthritis (RA) from the perspective of those who experience the disease. A particular emphasis at this workshop was placed on fatigue, but other areas included well-being, real-time assessment, patient priorities, and needs in early and late disease. Through a series of overview presentations, discussion groups, and plenary sessions, workshop participants (who included 19 patients) clarified what is known and what are the outstanding issues for future research. The importance of further work on clarifying the validity of fatigue measurements in RA has been confirmed, and with at least one suitable instrument available there will be strong pressure to include fatigue in a redefined core set of outcome measures in RA. In the other 4 areas covered there are important issues that can be addressed by enquiry and experiment and that together provide a challenging research agenda. At the final plenary session the OMERACT conference endorsed, by a large majority, the proposal that fatigue may warrant consideration for inclusion in the OMERACT core set for RA. PMID- 16265713 TI - Attributes and skills of an effective musculoskeletal consumer. AB - The OMERACT 7 Effective Musculoskeletal Consumer Workshop brought together people with rheumatoid arthritis, healthcare professionals, and researchers to discuss what they thought made a musculoskeletal consumer effective at managing their disease. Preliminary work before OMERACT provided a draft list of potential characteristics of an effective consumer. Participants at the workshop provided feedback about the list including relevance, missing items, format, and language. The feedback provided was useful and will be incorporated into a revised list to aid in the development of an instrument to measure health consumer effectiveness. PMID- 16265714 TI - Outcome measures in psoriatic arthritis. AB - Recent advances in biologic therapies have provided hope for patients with psoriatic arthritis (PsA). However, studies have been hampered by the lack of acceptable and validated outcome measures. This article reviews outcome measures used in the assessment of both skin and joints in PsA, and provides a summary of the Psoriatic Arthritis Workshop during OMERACT 7. A set of domains to be included in the assessment of patients with PsA was derived, and a research agenda was developed. PMID- 16265716 TI - Can hand assessments designed for persons with scleroderma be valid for persons with rheumatoid arthritis? PMID- 16265715 TI - Fibromyalgia syndrome. AB - The objectives of the first OMERACT Fibromyalgia Syndrome (FM) Workshop were to identify and prioritize symptom domains that should be consistently evaluated in FM clinical trials, and to identify aspects of domains and outcome measures that should be part of a concerted research agenda of FM researchers. Such an effort will help standardize and improve the quality of outcomes research in FM. A principal assumption in this workshop has been that there exists a clinical syndrome, generally known as FM, characterized by chronic widespread pain typically associated with fatigue, sleep disturbance, mood disturbance, and other symptoms and signs, and considered to be related to central neuromodulatory dysregulation. FM can be diagnosed using 1990 American College of Rheumatology criteria. In preparation for the workshop a Delphi exercise involving 23 FM researchers was conducted to establish a preliminary prioritization of domains of inquiry. At the OMERACT meeting, the workshop included presentation of the Delphi results; a review of placebo-controlled trials of FM treatment, with a focus on the outcome measures used and their performance; a panel discussion of the key issues in FM trials, from both an investigator and regulatory agency perspective; and a voting process by the workshop attendees. The results of the workshop were presented in the plenary session on the final day of the meeting. A prioritized list of domains of FM to be investigated was thus developed, key issues and controversies in the field were debated, and consensus on a research agenda on outcome measure development was reached. PMID- 16265718 TI - The arthritis of familial Mediterranean fever. PMID- 16265719 TI - Effects of beta-endorphin and naloxone on motility of cooled equine spermatozoa. PMID- 16265720 TI - Cooling rate, centrifugation and percoll selection in equine semen stored at +4 degrees C. PMID- 16265721 TI - Composition of fractionated stallion ejaculates. PMID- 16265722 TI - Immunocastration in stallions: effect on spermatogenesis and behaviour. PMID- 16265723 TI - Stallion spermatozoa membrane phospholipid dynamics following dietary n-3 supplementation. PMID- 16265724 TI - Effect of exercise on scrotal surface temperature in the stallion. PMID- 16265725 TI - [International study on testicular cancer: EORTC 30983: randomized phase II/III study of Taxol-BEP versus BEP in patients with Intermediate Prognosis Germ Cell Cancer]. PMID- 16265727 TI - Seminal plasma improves fertility of frozen equine semen. PMID- 16265726 TI - The effect of sodium hyaluronate on sperm function of cryopreserved equine spermatozoa. PMID- 16265728 TI - Evaluation of a cushioned centrifugation technique for processing equine semen for freezing. PMID- 16265729 TI - [Congress splitter]. PMID- 16265730 TI - Prepackaging of equine semen in goblets prior to cryopreservation. PMID- 16265732 TI - Xenografting of degenerate stallion testis onto a mouse host does not rescue the testicular degeneration phenotype. PMID- 16265731 TI - Effects of cooling stallion semen for 24 h before freezing on fertility rates. PMID- 16265733 TI - Seminal plasma proteins and semen characteristics in relation with fertility in the stallion. PMID- 16265734 TI - Influence of repeated artificial insemination (IS) on functional properties of uterine neutrophils of mares. PMID- 16265735 TI - Does equine sperm concentration influence the sperm migration to the oviducts? PMID- 16265736 TI - Effects of hemospermia on seminal quality. PMID- 16265738 TI - Relations between low rates of in vitro fertilization and induction of sperm acrosome reactions by zona pellucida in the equine species. PMID- 16265737 TI - Reproductive parameters of Mini-Shetland stallions in north Germany. PMID- 16265739 TI - The in vitro response of equine epididymal and ejaculated spermatozoa to caffeine. PMID- 16265740 TI - Effects of different extenders on sperm parameters and fertility of equine cooled semen. PMID- 16265741 TI - Effect of antibiotics on viability and fertility of equine semen cooled to 5 degrees C. PMID- 16265742 TI - Effect of extender and method of preservation on motility of cooled stallion spermatozoa. PMID- 16265743 TI - Effects of hyaluronic acid supplementation on motility of stallion spermatozoa after cryopreservation. PMID- 16265744 TI - Improvement of stallion semen post-thaw motility with the association dimethyl formamide and methyl formamide as cryoprotectors. PMID- 16265745 TI - Evaluation of acrosomal integrity of stallions cryopreserved with amides and glycerol. PMID- 16265746 TI - Cryopreservation of stallion semen collected from good and poor freezers using a directional freezing device (Harmony CryoCare--Multi Thermal Gradient 516). PMID- 16265747 TI - Effects of cushioned centrifugation technique on sperm recovery and sperm quality in stallions with good and poor semen freezability. PMID- 16265748 TI - Optimizing pregnancy rates using frozen-thawed equine semen. PMID- 16265749 TI - Evaluation of slow cooling after centrifugation and glycerol addition at 22 degrees C versus direct freezing of semen in stallions with good and poor sperm longevity. PMID- 16265750 TI - Glycerol in semen extender is a limiting factor in the fertility in asine and equine species. PMID- 16265751 TI - Equine seminal plasma proteins related with fertility. PMID- 16265752 TI - Inseminate components are modulating the chemotactic activity of uterine polymorphonuclear granulocytes (PMN) of mares. PMID- 16265753 TI - Effects of insemination volume and concentration of spermatozoa on post breeding endometritis and pregnancy rates in mares. PMID- 16265754 TI - Protein profile of equine seminal plasma: correlation to semen freezability. PMID- 16265755 TI - Improvement of the freezability of sex-sorted stallion spermatozoa. PMID- 16265756 TI - Effect of storage and temperature on in vitro stallion sperm parameters and fertility rate. PMID- 16265757 TI - Abstracts of the 57th Congress of the German Society of Urology, Dusseldorf, Germany, 21-24 September 2005. PMID- 16265758 TI - Abstracts of the American Society for Therapeutic Radiology and Oncology 47th Annual Meeting, October 16-20, 2005, Denver, Colorado, USA. PMID- 16265760 TI - [Acute abdomen -- diagnosis under analgesia]. PMID- 16265759 TI - [Reorganization of university clinics--Chirurg (2005) 76:68-70]. PMID- 16265761 TI - [Progress in the study of administration of drugs of the inner ear]. PMID- 16265762 TI - To have moral authority we first need to show that we can be critical of our own values. PMID- 16265763 TI - Yes, the system is more generous, but the cost in stress and uncertainty is wholly inordinate. PMID- 16265764 TI - Search for AIDS vaccine at risk because of lack of interest and funding. PMID- 16265765 TI - A medical miracle, but for how long? PMID- 16265766 TI - Clinical challenge. Plus ca change... PMID- 16265767 TI - HIV/AIDS and homelessness, Part 1: background and barriers to care. AB - Co-occurrence of homelessness and HIV/AIDS poses a complex and multidimensional challenge to the health care provider's clinical and system integration skills. Existing data support the high prevalence of HIV/AIDS among homeless persons and a high percentage of persons living with HIV/AIDS being either homeless or at imminent risk for homelessness. There are special considerations and challenges health care providers may face in caring for homeless persons with HIV/AIDS. An integrated, flexible, interdisciplinary, community-based system of care addressing the full array of medical, psychiatric/substance abuse, and housing services would optimize clinical care for this population. Areas that deserve particular attention include HIV/AIDS prevention, access to comprehensive HIV and health care, use of antiretroviral therapy, and adherence to treatment. Research is needed to better understand the multifaceted needs of this population and to develop prevention and treatment strategies applicable to daily clinical care. PMID- 16265768 TI - Ritonavir-boosted protease inhibitors, Part 2: cardiac implications of lipid alterations. AB - Ritonavir-boosted protease inhibitor (PI) regimens have provided substantial benefits in the treatment of HIV/AIDS, resulting in improved clinical outcomes. However, treatment toxicities often affect adherence and may influence outcomes. Dyslipidemia is highly prevalent in many patients who receive a boosted PI regimen. The mechanism underlying dyslipidemia is probably multifactorial and may differ among the individual PIs. The prevalence of cardiovascular disease and events appears to be higher in patients treated with boosted PIs, and as long term survival increases in HIV-infected persons, the long-term effect of dyslipidemia becomes a growing concern. The practitioner must consider the possibility of these adverse effects when choosing the antiretroviral regimen that best suits each patient. Until future studies define the optimal approach, an evaluation of cardiovascular risk factors and treatment of those risk factors according to evidence-based guidelines are warranted. PMID- 16265769 TI - Editorial comment: boosted PIs and metabolism--it's not just the ritonavir. PMID- 16265770 TI - Advances in HIV treatment: the 3rd International AIDS Society Conference. PMID- 16265771 TI - Inflammation as a cause of functional bowel disorders. PMID- 16265772 TI - Heterotopic gastric mucosa in the cervical esophagus: could this play a role in the pathogenesis of laryngopharyngeal reflux in a subgroup of patients with posterior laryngitis? AB - OBJECTIVE: Acid secretion produced by a heterotopic gastric mucosal patch (HGMP) in the proximal esophagus, instead of gastric acid, may be responsible for laryngopharyngeal reflux (LPR), passing the upper esophageal sphincter. The aim of this study was to investigate the prevalence of HGMP in the proximal esophagus in patients with posterior laryngitis indicating the presence of LPR in comparison with a control group and to elucidate the possible role of this lesion in the pathogenesis of LPR. MATERIAL AND METHODS: A total of 36 consecutive patients with posterior laryngitis diagnosed on laryngoscopic examination were enrolled in the study. Esophagoscopy and ambulatory 24-h intra-esophageal dual probe pH monitoring were performed in all patients. During endoscopy, special attention was paid to the proximal part of the esophagus, and the proximal electrode for pH monitoring was placed in this region under endoscopic view. The control group comprised 660 consecutive patients who had undergone upper gastrointestinal endoscopy for the usual indications. When HGMP was found, biopsies were taken for histological confirmation. RESULTS: HGMP was detected in 5 out of 36 patients. One out of five patients with patches was excluded from the study because the histopathology of this patient's patch revealed antral-type mucosa, which is not capable of acid secretion. Thus a total of 35 patients were included in the study, yielding a HGMP prevalence of 11.4% (4/35). Compared with the prevalence of the control group (1.6%), a significant difference was observed (p<0.005). pH monitoring showed that 45.4% of the patients had abnormal proximal acid reflux. All of four HGMP (+) patients with posterior laryngitis revealed significantly higher abnormal proximal reflux compared to the patients without patches (p<0.05). CONCLUSIONS: This first preliminary study may suggest that HGMP in the cervical esophagus could play a role in the pathogenesis of LPR, at least in a minor group of patients with posterior laryngitis, depending on its capability to produce acid in situ, although isolated proximal reflux could not be demonstrated. This finding may need to be supported by further studies with larger patient populations and using acid stimulation tests. PMID- 16265773 TI - Long-term morpho-functional development of Helicobacter pylori-induced gastritis in Mongolian gerbils. AB - OBJECTIVE: Epidemiological studies have shown that Helicobacter pylori infection with associated chronic gastritis is the main risk factor for development of gastric cancer. The aim of this study was to investigate the long-term development of H. pylori-induced gastritis in Mongolian gerbils in terms of morphology, gastrin secretion, epithelial proliferation and gene expression of pro-inflammatory cytokines. MATERIAL AND METHODS: A total of 133 gerbils were inoculated with H. pylori and 62 served as controls. The gerbils were killed at different time-points between 6 and 94 weeks after inoculation. Serum concentrations of anti-H. pylori IgG and gastrin were determined by enzyme-linked immunoabsorbent assay (ELISA) and radioimmunoassay (RIA), respectively. Epithelial proliferation was evaluated immunohistochemically after labeling with 5-bromo-2'-deoxy-uridine. Gene expression of beta-actin, interleukin-1beta (IL 1beta) and tumor necrosis factor-alpha (TNF-alpha) were measured by real-time reverse transcriptase-polymerase chain reaction (RT-PCR). Histological parameters of gastritis were assessed semiquantitatively and expressed as a "gastritis score". RESULTS: Serum concentrations of anti-H. pylori IgG and gastrin increased over time. Epithelial proliferation in the antrum was increased 6 weeks after inoculation, followed by increased proliferation in the corpus 32 weeks after inoculation. Gene expression of IL-1beta and TNF-alpha were increased in H. pylori-infected gerbils. Beta-actin was not a reliable endogenous control for RT PCR. With time, gastritis expanded from the antrum to the corpus and the gastritis score increased to reach a peak 32 weeks after inoculation. Pseudopyloric metaplasia (loss of specialized cells) was a characteristic feature in the corpus mucosa. Gastric ulcers, but neither dysplasia nor carcinoma, were observed during 94 weeks of infection. CONCLUSIONS: Long-term H. pylori infection in Mongolian gerbils led to progressive gastritis, glandular atrophy, hypergastrinemia, increased epithelial proliferation and elevated gene expression of pro-inflammatory cytokines. PMID- 16265774 TI - Postprandial water intake inhibits gastric antral motility with increase of cholecystokinin in humans. AB - OBJECTIVE: The effects of postprandial water intake on the gastrointestinal tract have not been systematically investigated in humans. MATERIAL AND METHODS: In 8 healthy volunteers, the gastric antral pressure was measured with a strain gauge transducer, while the esophageal and lower esophageal sphincter pressures were measured with an infused catheter with a Dent sleeve. The esophageal pH at 5 cm above the lower sphincter was measured with a microglass electrode. A standard test meal (560 kcal) was eaten and 500 ml water was ingested 1 h later. The plasma cholecystokinin level was assessed at 4-min intervals. As a control, the same study was done on another day with sham water intake. RESULTS: At 4 min after water intake, there was a significant decrease in gastric antral motility and a significant increase in the plasma cholecystokinin level. Water intake also significantly increased the occurrence of gastroesophageal reflux. CONCLUSIONS: The rapid increase in cholecystokinin after water intake may be initiated by a feedback mechanism related to inflow of fatty chyme into the duodenum that inhibits gastric antral activity. PMID- 16265776 TI - Skip inflammation of the appendiceal orifice: a prospective endoscopic study. AB - OBJECTIVE: The purpose of the study was to evaluate the incidence of discontinuous inflammation of the appendiceal orifice in patients undergoing colonoscopy for diagnosis or surveillance of colonic disease. MATERIAL AND METHODS: Consecutive and unselected patients subjected to colonoscopy over a 3 year period were included in a prospective study. Biopsies were taken within 2 cm of the orifice of the appendix, from the caecum and from predefined colonic segments. Discontinuous inflammation of the appendiceal orifice was defined as an area of macroscopic inflammatory changes distinct from a normal caecum of ascending colon. The biopsies were graded histologically for the presence and severity of inflammation by a pathologist without knowledge of the endoscopic findings. RESULTS: A total of 271 patients were included. The final diagnoses were: ulcerative colitis (UC) (83 patients), Crohn's disease (CD) (54), indeterminate colitis (12), irritable bowel syndrome (IBS) (54), microscopic colitis (15) and other disease (53). Endoscopic discontinuous inflammation of the appendiceal orifice was found in 27% (95% CI: 17-38%) of patients with UC, 24% (95% CI: 13-39%) with CD, 40% (95% CI: 12-74%) with indeterminate colitis, 8% (95% CI: 0-36%) with microscopic colitis, 10% (95% CI: 3-24%) of patients with IBS and in 9% (95% CI: 2-021%) of other diseases (p<0.05). A correlation was found for endoscopic and histological discrimination between normal and inflamed mucosa (p<0.001). However, in 24% of patients, endoscopic inflammation was without histological signs of inflammation, primarily in an otherwise normal colon. CONCLUSIONS: Discontinuous inflammation of the appendiceal orifice is common in patients with IBD irrespective of clinical activity. However, patients with otherwise normal colon may also show congestion of this area without or with minimal microscopic inflammation. PMID- 16265775 TI - Natural history of transglutaminase autoantibodies and mucosal changes in children carrying HLA-conferred celiac disease susceptibility. AB - OBJECTIVE: The natural history of the appearance and fate of transglutaminase autoantibodies (TGAs) and mucosal changes in children carrying HLA-conferred celiac disease (CD) risk remains obscure. The aim of this study was to investigate the sequence of events leading to overt CD by retrospective analysis of TGA values in serum samples collected frequently from genetically susceptible children since birth or early childhood. MATERIAL AND METHODS: A total of 1101 at risk children were recruited in the study. A duodenal biopsy was recommended to all TGA-positive children and performed if parental consent was obtained. RESULTS: During up to 8 years of follow-up, 35 of the cohort children developed TGAs, the youngest at age 1.3 years. After age 1.3 years the annual TGA seroconversion rate was constantly around 1% at least until age 6 years. However, 18 of the 35 TGA-positive children (51%) lost TGAs, without any dietary manipulation. A further 7 children were IgA deficient; of these children, 2 developed IgG antigliadin antibodies (IgG-AGA). Only 13 of the 21 children (62%) who had duodenal biopsies had villous atrophy. The time that passed since emergence of TGAs failed to predict the biopsy findings. Only one of the children with TGAs and both of the IgA-deficient children with IgG-AGA had noticeable abdominal symptoms. CONCLUSIONS: TGAs appear in children at a constant rate after 1 year of age until at least the age of 6 years. Over half of the children loose TGA without gluten exclusion, challenging TGA positivity-based CD prevalence estimates. In symptom-free children, a requirement of two consecutive TGA positive samples taken >or=3 months apart before performing a duodenal biopsy might diminish the number of unnecessary intestinal biopsies. PMID- 16265777 TI - A prospective, open-label trial of 6-thioguanine in patients with ulcerative or indeterminate colitis. AB - OBJECTIVE: 6-thioguanine (6-TG) has emerged as a promising therapeutic alternative in patients with Crohn's disease intolerant or resistant to azathioprine (AZA) and/or 6-mercaptopurine (6-MP). The aim of the present study was to evaluate the safety and efficacy of 6-TG in patients with ulcerative colitis (UC) or indeterminate colitis (IC) intolerant or resistant to AZA/6-MP. MATERIAL AND METHODS: Twenty patients with an acute flare, steroid-dependent or steroid-refractory disease attending our outpatient department were included in the study. Measurement of 6-TG nucleotide levels was done to check compliance. Complete, partial and non-response were defined by means of the clinical activity index and the daily steroid demand. Secondary outcome parameters included changes in cumulative steroid doses, C-reactive protein (CRP) levels, and an endoscopic score. RESULTS: Out of 20 patients 4 were excluded owing to noncompliance; 2/16 compliant patients (13%) had to be prematurely withdrawn because of adverse events, which ceased upon drug discontinuation. By per-protocol analysis, 5/14 patients (36%) were complete, 6/14 (43%) partial and 3/14 (21%) non-responders. In addition to the reduction of the cumulative steroid dose over 3 months, CRP decreased in the study population and the endoscopic score decreased in treatment responders. CONCLUSIONS: Treatment with 6-TG was effective in patients with UC or IC previously intolerant or resistant to AZA/6-MP. Future work is needed to define a subpopulation of patients at low risk for its potential hepatotoxicity, which we assume will benefit from 6-TG. PMID- 16265778 TI - Insulin-like growth factors (IGFs) and IGF binding proteins in active Crohn's disease treated with omega-3 or omega-6 fatty acids and corticosteroids. AB - OBJECTIVE: Catabolism and growth impairment are well-known complications of inflammatory bowel disease (IBD). This may be caused by the disease activity itself and/or the medical treatment, and both may lead to changes in the growth hormone (GH)/insulin-like growth factor I (IGF-I) axis. The aim of the present study was to examine the effects of enteral nutrition, Impact Powder, as adjuvant therapy to corticosteroid treatment on changes in the GH/IGF-I axis in patients with Crohn's disease (CD). MATERIAL AND METHODS: The patients were randomized to 3-IP (omega-3-fatty acid (FA), 3 g/day) or 6-IP (omega-6-FA, 9 g/day). Changes in total IGF-I (tIGF-I) and total IGF-II (tIGF-II), free IGF-I (fIGF-I), IGF binding proteins (IGFBP-1 and IGFBP-3), IGFBP-3 protease activity and insulin levels were examined in 31 patients with active CD (CDAI: 186-603) during treatment with prednisolone (40 mg for 1 week) and tapering the dose by 5 mg/week. Clinical and biochemical markers of inflammation were studied at day 0, and after 5 and 9 weeks. RESULTS: There were no differences at baseline between the two groups. During the treatment period, tIGF-I, fIGF-I and IGFBP-3 increased significantly in both groups compared to baseline (p<0.05) without differences between the groups. Insulin and IGFBP-1 showed no significant changes throughout the treatment period. CONCLUSIONS: There was no difference between 3-IP and 6-IP as adjuvant enteral nutrition on the GH/IGF-I axis. The changes observed in the GH/IGF-I axis are in line with previously published studies and may be explained by corticosteroid treatment; however, we cannot exclude an additional effect of omega3-/omega6 FA as adjuvant enteral nutrition. PMID- 16265779 TI - Routine application of the proton-pump inhibitor pantoprazole in patients with gastric lymphoma undergoing chemotherapy. AB - OBJECTIVE: The stomach is the most common site of origin for extranodal lymphomas. While resection has played a major part in the management of such patients in the past, in recent years there has been a change towards organ conserving therapies. However, the risk of perforation and bleeding in patients undergoing organ-conserving therapy has been used as an argument against primary application of chemotherapy. In this article, we present our experience with the prophylactic use of oral pantoprazole. MATERIAL AND METHODS: All patients undergoing chemotherapy for gastric lymphoma at our institution were prophylactically given oral pantoprazole (2x40 mg) for the duration of chemotherapy. Compliance with intake of the proton-pump inhibitor (PPI) was assessed at every visit for application of chemotherapy and at routine blood counts taken 10-14 days after each cycle of treatment by direct questioning of the patient. RESULTS: A total of 82 patients (median age 69 years, range 33-93) received chemotherapy for gastric lymphoma: 51 had diffuse large B-cell lymphoma (DLBCL), 24 had mucosa-associated lymphoid tissue (MALT) lymphoma and 7 had DLBCL+MALT lymphoma. Compliance with intake of the PPI was excellent, as only two patients reported irregular intake and only one patient refused regular medication with pantoprazople. All patients responded to chemotherapy, with 74 (90%) achieving complete remission and 8 (10%) partial remission. After a median follow-up time of 50 months (range: 9-84 months) only one of these 82 patients (1.22%), the patient who discontinued intake of pantoprazole, died from gastric perforation, while none of the other patients had gastrointestinal bleeding or perforation. CONCLUSIONS: Judging from these data, continuous PPI intake is feasible and has a high rate of compliance. In the absence of randomized trials, routine application of oral pantoprazole in patients given chemotherapy for gastric lymphoma, especially DLBCL, appears to be a reasonable approach. PMID- 16265780 TI - Is it long-term continuous drinking or the post-drinking withdrawal period that triggers the first acute alcoholic pancreatitis? AB - OBJECTIVE: Unlike patients with alcoholic hepatitis, patients with acute alcoholic pancreatitis seldom come into the hospital in an intoxicated state. Long-term history of heavy drinking induces increases in the serum pancreatic enzymes and pancreatitis-associated protein profiles during the withdrawal period. The aim of this study was to investigate the role of withdrawal in triggering acute alcoholic pancreatitis by studying the time-course of development of the first symptoms of the first acute alcoholic pancreatitis. MATERIAL AND METHODS: One hundred patients (85 M, 15 F, mean age 46, range 18-73 years) with the first acute alcoholic pancreatitis were asked three different questions in an attempt to clarify the same issue: Had you already stopped continuous drinking before the start of the acute abdominal pain that later led to hospitalization? Had you already stopped continuous drinking before you started to experience nausea or vomiting? How many hours after taking the last drop of alcohol did you start to feel pain (0 h, 1<6 h, 7-12 h, 13-24 h, 25-48 h, >48 h)? The amount of alcohol consumed was evaluated 1) during the past week and 2) during the past 2 months. The severity of the pancreatitis was assessed by serum C-reactive protein concentration, presence of necrosis, the development of pancreatic complications and the length of stay in hospital and in the intensive care unit. RESULTS: Eighty-five patients were able to respond to the questions. Of these, 69% had developed pain and 91% nausea/vomiting only after they had already stopped continuous drinking. Whereas 29% of the patients developed some symptoms before stopping drinking, the majority of the patients developed symptoms during the first day after cessation (43%) or later (28%), mainly during the second day of cessation of drinking. In both the univariate analysis and the multivariate analysis the timing of the symptoms was dependent on the amount of alcohol consumed during the previous 2 months and in the past week. CONCLUSIONS: In the majority of patients with first acute alcoholic pancreatitis, the symptoms begin during the early withdrawal period. The withdrawal period might be more important than previously emphasized in the development of acute alcoholic pancreatitis. PMID- 16265781 TI - Patients with serum IgA endomysial antibodies and intact duodenal villi: clinical characteristics and management options. PMID- 16265783 TI - Treatment-resistant lingual Crohn's disease disappears after infliximab. AB - Oral manifestations of Crohn's disease (CD) are not uncommon, but they can be difficult to diagnose and treat. We describe a patient with long-standing CD and a lingual ulcer, which we attributed to CD. The oral lesions were unresponsive to conventional therapy such as steroids, mesalamine, and other topical agents. There was an excellent response to infliximab, a chimeric monoclonal antibody to tumor necrosis factor (TNF-alpha). In the context of this case we discuss the various differential diagnoses. Furthermore, we report on different therapeutic options and their effectiveness. Oral manifestations of CD, which are refractory to systemic steroids and mesalamine, show an excellent response to infliximab. PMID- 16265782 TI - Colon stenosis in a patient with ulcerative colitis as a manifestation of mixed mullerian tumor of the peritoneum. AB - We report the case of a 56-year-old woman followed for ulcerative colitis since 1987. In 1993, she underwent a total hysterectomy with bilateral salpingo oophorectomy. In 1996, a narrowing of the lumen at 20 cm of the anal margin was observed and she was put under endoscopic observation. In 1998, a mass in the stricture was observed, with significant narrowing of the lumen. Histologic examination revealed a biphasic tumor with features of malignant mixed mullerian tumor. Five years after surgery and chemotherapy with paclitaxel and carboplatinum, the patient remains well. PMID- 16265784 TI - AMCA President's address delivered at the AMCA Annual Conference April 4, 2005, Vancouver, British Columbia, Canada. PMID- 16265785 TI - Carbonic anhydrase type II deficiency. PMID- 16265786 TI - The cost of HIV medication adherence support interventions: results of a cross site evaluation. AB - The objective of this study was to determine the direct cost of HIV adherence support programmes participating in a cross-site evaluation in the US. Data on the frequency, type, and setting of adherence encounters; providers' professions; and adherence tools provided were collected for 1,122 patients enrolled in 13 interventions at 9 sites. The site staff estimated the average duration of each type of encounter and national wage rates were used for labour costs. The median (range) adherence encounters/year among interventions was 16.5 (4.3-104.6) per patient; encounters lasted 24.6 (8.9-40.9) minutes. Intervention direct cost was correlated with the average frequency of encounters (r = 0.57), but not with encounter duration or providers' professions. The median direct cost/month was 35 dollars(5 dollars-58 dollars) per patient, and included direct provider costs (66%); incentives (17%); reminders and other tools (8%); and direct administrative time, provider transportation, training, and home delivery (9%). The median direct cost/month from a societal perspective, which includes patient time and travel costs, was 47 dollars(24 dollars-114 dollars) per patient. Adherence interventions with moderate efficacy costing < or =100 dollars/month have been estimated to meet a cost-effectiveness threshold that is generally accepted in the US. Payers should consider enhanced reimbursement for adherence support services. PMID- 16265787 TI - Lowering costs by encouraging healthy behaviors. PMID- 16265788 TI - RNA inhibition of beta-secretase reverts AD in mice. PMID- 16265789 TI - Prime time for multiple sclerosis genetics. PMID- 16265790 TI - Channelling treatment for neuropathic pain. PMID- 16265791 TI - Japanese encephalitis outbreak in India. PMID- 16265792 TI - In search of the smallest infectious prion. PMID- 16265793 TI - New mouse model with Down's syndrome phenotypes. PMID- 16265794 TI - Protein culprit found for transverse myelitis? PMID- 16265795 TI - Ability to tune out irrelevant information is impaired in AD. PMID- 16265796 TI - Adaptation to bereavement. AB - Much of the literature on the effects of conjugal bereavement has focused on the detrimental effects of losing a spouse. Although it is very important to be aware of the emotional, physical, and social problems often associated with bereavement, these difficulties are not universal accompaniments of grief. Accumulating evidence suggests that resilience in the face of bereavement is the norm rather than an exception. This article uses a multidimensional model of bereavement adaptation and reviews literature on multiple paths to resilience among bereaved individuals. Each dimension of adaptation is illustrated with quotes from participants of the San Diego Widowhood Study. PMID- 16265797 TI - Bereavement and the brain: invitation to a conversation between bereavement researchers and neuroscientists. AB - A recent development by neuroscience is neuroimaging, a method of looking into the "black box" of the brain while people are feeling, doing, and thinking in real time. The first fMRI study of bereavement has recently been published, and the present article summarizes it in non-specialist language, focusing on its theoretical and clinical applications. In an attempt to bridge the gap between bereavement researchers and neuroscientists, the author discusses how these two fields could assist each other in forwarding both fields. Three current debates in the field of bereavement research are outlined, including (a) adaptation in the normal grief process, (b) complicated grief vs. resilience, and (c) meaning making vs. return-to-baseline models of bereavement. The potential contribution of neuroscientific data to these debates is discussed in several hypothetical examples. These examples stimulate thinking about the reciprocity between 2 questions: What can bereavement teach us about the brain? and What can the brain tell us about bereavement? This article is designed to provide enough background for investigators who are primarily concerned with the brain and those primarily concerned with bereavement to open a dialogue between both of these fields. PMID- 16265798 TI - The experience of Chinese bereaved persons: a preliminary study of meaning making and continuing bonds. AB - This study explores the bereavement process of Chinese persons in Hong Kong, with the focus on how they make meaning of the death as well as how they maintain a bond with the deceased. A review of video- and audiotapes of 52 bereaved persons in bereavement counseling pointed to how these concepts are reflected in key themes that appeared throughout these interviews. The way this bereaved population found meaning in the death was reflected in how they understood the cause of death, their observations at the death moment, their understanding of the life of the deceased after the death, the life of the bereaved after the death, and the burial and mourning rituals that were followed. The continuing bond the bereaved felt with the deceased was experienced by the bereaved as either initiated by the deceased, for example by the deceased appearing in dreams or initiated by the bereaved themselves in their talking with the deceased. These 2 aspects of the bereavement process seem to be similar to those found in other societies, but the ways in which they were manifested were unique to the Chinese culture. Based on these findings, the authors conclude with implications for intervention with this bereaved population. PMID- 16265799 TI - Happiness and death distress: two separate factors. AB - The aim of the present study was to explore the relationship between happiness and death distress (death anxiety, death depression, and death obsession) in 275 volunteer Kuwaiti undergraduates. They responded to the Oxford Happiness Inventory, the Death Anxiety Scale, the Arabic Scale of Death Anxiety, the Death Depression Scale-Revised, and the Death Obsession Scale. Gender differences were significant on all 5 scales, with women showing a lower mean score of happiness and a higher mean score for the death distress. All the correlations between happiness and the death distress scales were non-significant except one pertaining to happiness and death depression (negative) in women. Two oblique factors were extracted: death distress and happiness. Therefore, these constructs represent 2 distinct and independent factors. PMID- 16265800 TI - Predictors of strong sense of coherence and positive attitudes to physical education in adolescents. AB - AIMS: The aim of this study was to find variables related to positive attitudes to physical education (PE) and strong sense of coherence (SOC) among adolescents. METHODS: The study included three parts: seven physical tests; a questionnaire which included ways of living, attitudes to PE, and subjective health, the 13 question version of SOC; and information on every student's grades. The study group comprised 301 teenagers (131 girls and 170 boys, aged 16-19 years) attending upper secondary school. Positive odds ratio was used in the logistic regression analyses with SOC and attitudes to PE as dependent variables. RESULTS: Variables related to positive attitudes to PE were strong SOC, high physical capacity, high leisure-time physical activity (PA), high grades in PE, and little time spent watching TV. Variables related to strong SOC were positive attitudes to PE, high grades in PE, very good subjective health, and feeling comfortable in school. Highest physical capacity, highest mean grades, and highest grades in PE were found among adolescents who reported exercise four times or more per week. CONCLUSIONS: An interrelation between attitudes to PE and SOC was shown. The relation between positive attitudes to PE and high scores in SOC indicated that past experiences of PA and PE could contribute to the development of SOC, and actual levels of SOC could influence the persistent attitudes to PE and be important for lifelong PA. One means of identification of favourable or unfavourable health behaviour among young people might be through PA patterns, and relations between attitudes to PE and SOC. PMID- 16265801 TI - A model for cost-effectiveness analyses of smoking cessation interventions applied to a Quit-and-Win contest for mothers of small children. AB - AIMS: The first aim of this study was to develop a model that predicts health and economic consequences of smoking cessation in Sweden, striving to follow the methodological recommendations to reflect the societal perspective and to use the health measure quality-adjusted life-years (QALYs). The second aim was to apply the model estimates to a smoking cessation intervention. METHODS: A Markov cost effectiveness model was developed on smoking-related lung cancer, chronic obstructive pulmonary disease, and cardiovascular disease. Swedish primary data on medical treatment costs and quality-of-life weights were used, supplemented with secondary data on other societal effects and the disease and death risks. The model simulations were applied to a "Quit-and-Win" contest for mothers of pre school children. In total, 238 women participated, with 34 sustained tobacco-free at 12-month follow-up. RESULTS: The cost-effectiveness model estimates a gain of 0.34 to 0.55 QALYs (discounted 3%), and cost savings of SEK 20-35,000 per female quitter in the age range 15 to 49 years. The cost-effectiveness analyses of the intervention showed intervention costs per quitter of SEK 7,850, and intervention costs per life-years saved (YLS) (discounted 3%) of SEK 13,200. The cost-utility analysis demonstrated cost savings and a gain of 16 QALYs. CONCLUSIONS: The cost utility analysis estimated health gains and cost savings resulting from the "Quit and-Win" contest. As the model estimates on the differences in societal cost between smokers and quitters are considerable, many tobacco control programmes would result in cost savings. The construction of an optimal mix of tobacco control policies, however, demands incremental calculations on a range of programmes. PMID- 16265802 TI - Breastfeeding attitudes among counselling health professionals. AB - AIM: The aim of the study was to develop an instrument that can be used for accurate assessment of nurses' and midwives' attitudes toward breastfeeding in a group of midwives, maternity-nursing staff and postnatal nurses experienced in breastfeeding counselling. METHOD: An instrument based on WHO standards was developed to measure breastfeeding attitudes. In all, 168 healthcare professionals filled in the instrument. A factor analysis using maximum likelihood and varimax rotation was performed. Spearman's correlation was used to correlate factorial dimensions and self-described interest in breastfeeding. RESULTS: By means of factor analysis four factors were identified: the "regulating" factor focused on regulating the mothers' breastfeeding management, the "facilitating" factor focused on making it easy for mothers to manage their breastfeeding, the "disempowering" factor focused on giving advice, disregarding the needs of the mother being counselled, and the "breastfeeding antipathy" factor focused on insufficient, basic, breastfeeding knowledge and aversive reactions to breastfeeding. Midwives rated higher on the facilitating factor and breastfeeding antipathy factor and lower on the regulating factor than postnatal nurses. Breastfeeding interest was positively correlated with the facilitating factor, and negatively with the disempowering factor and the breastfeeding antipathy factor. CONCLUSION: This instrument provides a picture of health professionals' attitudes towards breastfeeding. Four factors were identified in order of importance: regulating, facilitating, disempowering, and breastfeeding antipathy factors. Harmful attitudes were identified and suggested a need for educational programmes to help health professionals to reconcile damaging values, in order to improve breastfeeding counselling. PMID- 16265803 TI - The return of eugenics. PMID- 16265804 TI - Role dilemmas among health-workers in cross-cultural patient encounters around dietary advice. AB - AIM: The aim of this paper is to explore Norwegian health workers' experiences from cross-cultural patient encounters, and how they understand and enact their role when meeting patients with Pakistani background to whom they give dietary advice related to type 2 diabetes. METHODS: Qualitative in-depth interviews have been performed with six hospital dietitians and six general practitioners in Oslo. RESULTS: The health workers consider themselves to be patient-centred and stress the importance of the two dimensions, empathy and equality. However, they often experience that patients want them to be more authoritarian, a way of acting that would be totally in disagreement with their convictions, although some occasionally do adopt an authoritarian style. More striking is that some health workers' moral engagement to involve and empower patients actually leads them to be authoritarian. For others, a fear of insulting the patient results in their advice being too diffuse. CONCLUSIONS: A possible explanation for such ways of responding to the patient may be that the health workers, in their articulation of patient-centredness, draw on a repertoire of social conduct that involves an effort to level out, or tacitly deny, hierarchic structures, and that this becomes more pronounced in cross-cultural encounters. Patient-centredness and empowerment are results of long ongoing processes in Western countries, based on ideals of equality and individual freedom. The results from this study indicate that these approaches may pose intricate dilemmas for the health workers in their cross-cultural encounters, and need further attention. PMID- 16265805 TI - When does human life begin? PMID- 16265806 TI - Long-term sick-listing among women in the public sector and its associations with age, social situation, lifestyle, and work factors: a three-year follow-up study. AB - BACKGROUND: Since 1997 the number of long-term sick-listed people in Sweden has increased dramatically, especially among women employed in the public sector. AIMS: The aim of this study was to investigate associations between age, social situation, lifestyle, work factors, and long-term sick-listing (> or =28 days), regardless of diagnosis, during a period of three years among women employed in the public sector. METHODS: Exposure information at baseline was recorded by a questionnaire. All new spells of sick listing (> or =28 days were consecutively reported from the employer for three years from baseline. In total 6,246 women from the public sector in Sweden answered the questionnaire (85% response rate); 5,224 were classified as having good or rather good health for working and were included. Of these, 918 persons had spells of sick-listing (> or =28 days during follow-up. RESULTS: Univariate and multivariate analyses for calculating relative risks (RR) were carried out. In the Cox regression model age (RR 1.4), strained financial situation (RR 1.3), obesity (RR 1.3), bullying (RR 1.5), physical demands at work higher than own capacity (RR 1.5), and mental demands at work higher than own capacity (1.2) remained risk indicators for long-term sick listing. CONCLUSION: This study suggests prevention of some work and lifestyle factors as general measures to reduce long-term sick-listing among women in the public sector. PMID- 16265807 TI - Cardiac rehabilitation services in Denmark: still room for expansion. AB - AIM: European cardiologists agree that cardiac rehabilitation (CR) should be offered as an integrated part of cardiac care, and CR guidelines have been published. The authors aimed to ascertain the potential for expanding CR coverage at hospitals in Denmark. METHOD: A cross-sectional questionnaire study was conducted among all hospitals receiving acute cardiac patients (n = 67). The response rate was 79%, with no differences according to catchment area, number of beds, or geographical location. The hospitals were classified as having full CR if all core components (physical training, psychosocial support, dietary counselling, smoking cessation, and pharmaceutical risk factor management) were available during each of three phases: (I) in hospital; (II) outpatient; and (II) community-based services. RESULTS: Many hospitals offered one or more of the CR components during phases I and II: physical training (77%; 77%), psychosocial support (89%; 79%), dietary counselling (85%; 89%), smoking cessation (94%; 68%), and clinical control by a physician (100%; 93%). The content varied greatly. Full phase I CR was offered at 57% (95% confidence interval (95% CI): 44-70%) of the hospitals and 47% (95% CI: 34-60%) offered full phase II CR. Phase III CR was very rare (2% (95% CI: 0-6%). The numbers of patients receiving CR was not registered. CONCLUSION: Marked progress was made in the 1990s in implementing CR; nevertheless, the services are far from fully expanded. Denmark has great potential for improving CR services, as do most other European countries. CR activities need to be registered at Danish hospitals. PMID- 16265808 TI - Can artificial techniques supply morally neutral human embryos for research? AB - Amidst controversy surrounding research on human embryos, biotechnology has conceived a substitute in the artificial human embryo. We examine the claim that novel embryos constructed artificially should be exempt from ethical restraints appropriate for research on embryos that come into being through natural processes. Morally relevant differences in intrinsic value depend on the sense in which the entity may be artificial, whether in regard to constituent matter, genetic or cellular form, generative means, or intended purpose. Considering each of these Aristotelian categories from a physicalist viewpoint, technology can achieve only limited degrees of artificiality because redesigned embryos still retain most of their natural features and relationships. From an essentialist viewpoint, the very limits of technology preclude the capability of manipulating the fundamental nature or essence of the individual who, even at the embryonic stage of life, cannot be made to be artificial through and through. A human may possess artificially contributed attributes but cannot be an artificial being. Classification of novel human organisms as artificial, therefore, is insufficient grounds by which to relinquish the principle that human moral status should be recognized for all living beings of human origin. In uncertain cases, at least the possibility of special human moral status should be considered present in organisms that are derived asexually, are developmentally defective, or are otherwise technologically altered. PMID- 16265809 TI - Clinical ethics dilemmas. PMID- 16265810 TI - Balint groups. PMID- 16265811 TI - Applying theological developments to bioethical issues such as genetic screening. AB - Catholic movements within the centre of Roman Catholic doctrine recently have discussed Trinitarian theology as applied to sciences, arts, economics, health and other social areas. We explore the possibilities Trinitarian theology offers to bioethical debate, concentrating particularly on genetic screening and testing. It is important therefore to analyse the philosophical implications of this approach onto the bioethical world, where much disagreement occurs on fundamental issues. It is Catholic basic teaching to recognize and see God's hand in plurality, not merely as a cliche and then doing what we feel is right, but to recognize how to live in a pluralistic world. We recognize, in agreement with these theologians, that in order for a Trinitarian mode of understanding to be used by those doing bioethical debate, there is a need to depart from fundamentalism. PMID- 16265812 TI - Recurrent vulvovaginal candidiasis. PMID- 16265813 TI - Infertility. PMID- 16265814 TI - The longevity of Baseball Hall of Famers compared to other players. AB - The authors compared the longevity of all baseball players alive at the time of their induction into the Baseball Hall of Fame with age-matched controls who were likewise alive at the time of the Hall of Famer's induction, and also matched them for career length, player position, and body-mass index, to assess if fame in sports is associated with increased longevity. Median post-induction survival for Hall of Famers was 5 years shorter than for noninducted players (18 vs. 23 years, respectively). In a second analysis, significantly more Hall of Famers died of cardiovascular or stroke causes than other players for whom cause of death was known. Baseball fame may have a hitherto unrecognized price. PMID- 16265816 TI - Seeking knowledge. PMID- 16265817 TI - Greetings from Nigeria. PMID- 16265818 TI - The inside dope. PMID- 16265819 TI - Prion biology in transfusion medicine: implications for lab testing. AB - Although eerily silent for many years after the recognition of scrapie in 1759, TSEs remained present within the genome of some mammals. Not since the mid-1950s when Dr. Carleton Gadjusek visited the Fore Indians of New Guinea to study kuru, however, has there been a more frenetic interest by governmental investigators. Certainly, the U.K. experience has heralded a renewed interest in TSEs due to the notoriety associated with younger subjects succumbing to a variant CJD traced to the ingestion of beef. Human TSEs and the potential for their transmission among and across species of mammals has also captured the attention of many. Yet, to date, there is no reliable antemortem test available to screen for infected animals or humans. Antibody-based assays are difficult to develop because most of them do not have specificity for the pathogenic form of prion protein. Whether or not prion testing efforts will change dramatically depends upon the incidence of disease. Some speculate a reduction in testing, because BSE incidence is waning since the adoption of remedial steps in the U.K. in 1989. Others remind us, however, of the long latency of prion diseases and of the recent observations of two patients who succumbed to vCJD after having received blood products from donors who subsequently died of vCJD. The growing incidence of CWD, combined with the emerging observation that as many as 26% of Alzheimer's patients may have been misdiagnosed--having died instead of prion disease--maintains pressure for legislators to adhere to the precautionary principle and support blood-donor exclusionary criteria, antemortem-test development, and pathogen removal from donated blood. The laboratorian can expect to see new tests for prion disease work their way into clinical-testing practice in the near future. In addition, the adoption of newer filtration technologies holds the promise of improved protection from transfusion-transmitted prion disease. PMID- 16265820 TI - Immunology in the 21st century: innovative tools in T-cell immunology. PMID- 16265821 TI - Prepare for a major disaster: the laboratory and beyond. PMID- 16265822 TI - Terrorism and emerging infections demand precaution. PMID- 16265823 TI - At SoftTech Health, automation is not just for testing anymore. PMID- 16265824 TI - I hope you dance. PMID- 16265825 TI - Is this lab duty bound to patient? PMID- 16265826 TI - Addressing management issues. PMID- 16265827 TI - Why we need passage of HR 1175. PMID- 16265828 TI - Effect of hydroxyethylstarch on renal function in cardiac surgery: a large scale retrospective study. AB - BACKGROUND: Recent reports indicated negative effects of hydroxyethylstarch (HES) on renal function. The goal of this large scale retrospective study was to detect whether there was an association between postoperative deterioration of renal function and the use of HES 200 kD, 0.5 DS in the cardiac surgery setting. METHODS: Retrospective analysis of daily collected data in 3124 patients who underwent coronary artery bypass and/or valvular surgery. Three groups were compared according to differences in fluid therapy: --GEL: gelatin was used as priming fluid of extracorporeal circulation (ECC) and for postoperative filling (n = 1276). --MIX: HES was used as priming fluid of ECC an gelatin was used for postoperative filling (n = 1008). --HES: HES was used as priming fluid of ECC and for postoperative filling (n = 840). MAIN RESULTS: There were no significant differences in postoperative serum creatinine concentrations between the 3 groups: GEL: 12,2 +/- 0,5 mg/l; MIX: 12,3 +/- 0.5 mg/l; HES: 12,3 +/- 0.6 mg/l. The need for postoperative extrarenal epuration was not significantly different between the 3 periods: GEL: 2,9%; MIX: 3,1%; HES: 3,8%. CONCLUSION: The use of HES 200 kD, 0.5 DS in cardiac surgery does not seem to be associated with a clinically significant deterioration of postoperative renal function. PMID- 16265829 TI - Conventional treatment or epidural blood patch for the treatment of different etiologies of post dural puncture headache. AB - Post dural puncture headache (PDPH) represents a complication of anesthesia (with an increased incidence in obstetric patients) or as the consequence of a diagnostic lumbar puncture. The aim of the present study was to evaluate the efficacy of the epidural blood-patch (EBP) versus the conventional medical treatment of post-anesthetic headaches also including the PDPH following a diagnostic puncture, a category of patients rarely referred to the anesthesia consultation in our hospital because it was believed that they might have equal benefit from conventional measures due to the smaller size of needles used. We studied in a prospective, randomized, double-blinded manner 32 obstetric and non obstetric patients with PDPH having the onset of the symptoms 24 hours before the inclusion in the study. The patients were randomly divided in two groups: group A (16 patients) receiving conventional treatment (oral and intravenous fluid replacement, non-steroidal anti-inflammatory drugs--NSAIDs--, caffeine) and group B (16 patients) in whom an epidural blood-patch was performed. The intensity of the headache was evaluated using a visual analogue scale (VAS) from 0-10, before, 2 hours and 24 hours after the EBP. There were no statistical differences concerning the demographic data and the cause of PDPH between the groups (p > 0.05). The intensity of PDPH was similar before performing the EBP (p > 0.05), with a value on VAS of 8.2 +/- 1,4. in group A and 8,0 +/- 1.6 in group B. Two hours after the treatment, the intensity of headache on VAS diminished extremely significant (p < 0.0001): in group B the value was 1.0 +/- 0,18 versus 8.2 +/- 1.4 in group A. The difference recorded after 24 hours remained statistically significant (p < 0.0001): the VAS scores were 0.7 +/- 0,16 and 7.8 +/- 1.2 respectively. The epidural blood patch represents the first choice treatment of PDPH no matter the etiology, being significantly superior to the conventional treatment which did not affect pain scores. In severe PDPH there is no reason to delay the EBP more than 24 hours after the diagnosis as all except two patients of the conventional treatment group required blood patching following the study period. PMID- 16265830 TI - The history of peripheral intravenous catheters: how little plastic tubes revolutionized medicine. PMID- 16265831 TI - New closed IV catheter system. PMID- 16265832 TI - QMDA, quality management and departmental administration. AB - Today, quality is part of our practice. Anaesthetists are involved in a medical science and want to be excellent at it. Anaesthetists are also involved in departmental organisation and are providers of care, a service to the patients. Quality is a method to obtain efficiency (efficacy and economical advantages). We consider our patients no longer as such, but as health-customers. Anaesthetists are health providers. The patients-customers have their own expectations and they compare the service between the different hospitals. On the other hand, health care is a public matter and in most countries depends on a governmental financing. Quality means efficacy, ethics and economics, including medico-legal aspects. But how can we use a quality and management concept (QMDA) in our daily practice? This is the subject of the paper. PMID- 16265833 TI - Metformin lactic acidosis and anaesthesia: myth or reality? AB - AIMS: To determine whether a causal or coincidental relationship is indicated in the literature between metaformin and lactic acidosis and to recommend clinical guidelines for the withdrawal of metformin prior to surgery. METHOD: A broad review of the literature related to metformin associated acidosis was carried out. (There are few publications specifically related to metformin treatment and anaesthesiology). RESULTS: When metformin-associated lactic acidosis occurs, a concurrent pathology or contraindication to the use of metformin is often found. Anaesthesia and surgery can generate or aggravate concurrent pathologies. CONCLUSION: Although no association has been shown between metformin and lactic acidosis under usual conditions of use, vigilance is required when metformin is used prior to surgery. The following clinical guideline is proposed: to withdraw (when possible) metformin 48 hours prior to surgery and to wait until the patient's biological and clinical parameters return to normal before reintroducing it. PMID- 16265834 TI - Echocardiography for management of low cardiac output state after axillary block in a dehydrated patient with aortic stenosis: case report. AB - We report an episode of transient low cardiac output in a dehydrated elderly woman having an undetected clinically significant aortic stenosis, after an axillary blockade. Cardiac echography was determinant for management of this patient. Events were considered to be from hemodynamic origin. PMID- 16265835 TI - ALFs strengthen community bonds. PMID- 16265837 TI - It's time for a health information revolution. PMID- 16265838 TI - Not-for-profit hospitals alter billing and collection practices for uninsured patients. Revised charity policies show negligible impact on bottom line. PMID- 16265839 TI - Country's health care quality up, peformance feedback down. Consumer-directed plans place less importance on quality, study says. PMID- 16265836 TI - Promoting memory retention. PMID- 16265840 TI - CBPP ridicules governors' ideas to reduce medicaid costs. Advocacy group claims proposals endanger low-income, disabled benes. PMID- 16265842 TI - Tourette syndrome: not just a tic disorder. AB - Although tics are considered the hallmark of Tourette syndrome, arguably tics may not be the only or primary presenting symptom. For many children diagnosed with Tourette syndrome irritability, frustration intolerance, hyperactivity, inattention, ritual behavior or other difficulties may have been present a number of years before the appearance of tics. Children with Tourette syndrome are often highly co-morbid with attention deficit-hyperactivity disorder, obsessive compulsive symptoms, and other related behavioral problems that should be detected and treated effectively. Therefore tics should not be the sole indicator or receive over emphasis in the detection and treatment of Tourette syndrome. PMID- 16265841 TI - Myotonic dystrophies. AB - Myotonic dystrophies or dystrophia myotonica (DM) is a clinical syndrome that includes myotonic dystrophy type 1 (DM1), myotonic dystrophy type 2 (DM2), myotonic dystrophy type 3 (DM3), and so forth. The terminology was recommended by the new nomenclature for myotonic dystrophies of an International Panel for Consensus. Previous studies have shown that DM1 is caused by the expansion of a cytosine-thymine-guanine (CTG) repeat in the DM protein kinase gene on chromosome 19, and DM2 is caused by an expansion of a cytosine-cytosine-thymine-guanine (CCTG) repeat in the zinc finger protein 9 (ZNF9) gene on chromosome 3. Because DM1 and DM2 have very similar clinical presentations, the diagnosis of these two disorders needs to be confirmed by molecular genetic analysis. Recently, DM3 was reported to include a multisystem myotonic disorder with frontotemporal dementia, and a linkage to chromosome 15q21-24. Although the age at onset, disease severity, and cerebral abnormality on a brain magnetic resonance spectrometry may correlate with the number of triplet repeats in the blood cells of DM1, it is too early to reach a conclusion. In Taiwan, the prevalence of DM1 is much lower than in Western countries. Previous studies have shown that the central nervous system symptomatology is correlated mainly with the white matter lesions in the brain MRI, but the CNS manifestations seem unrelated to the numbers of CTG triplet repeats in the blood cells. The inverse correlation between age at onset and CTG repeat length is significant only in patients with small expansions of about 100 250 triplet repeats. Transmission contraction of the repeat size is likely to occur in alleles with large repeats and is associated with paternal transmission. In congenital DM1, individual variability of muscle differentiation does occur, in spite of the same number of CTG repeats in the leukocytes. PMID- 16265843 TI - Lectinochemical studies on the binding properties of a toxic lectin (ricin) isolated from the seeds of Ricinus communis. AB - BACKGROUND: Ricin (RCA2 or RCA60) is a highly toxic heterodimeric protein found in the seeds of the castor plant Ricinus communis. It is a potential biohazard. In the present study, the fine specificity of ricin was defined. METHODS: The combining site of ricin was characterized by quantitative precipitin (QPA) and precipitin inhibition assays (QPIA). RESULTS: Of 31 glycoproteins and pneumococcus type XIV capsular polysaccharide tested, only twelve of them precipitated over 50% of the toxin N added, reflecting poor precipitability of the lectin with the compounds tested. This can be explained by only a single chain (B chain of the molecules) participating in binding. The blood group active glycoproteins after mild acid hydrolysis or Smith degradation, as well as sialic acid containing glycoproteins after removal of sialic acid, in general, had substantially increased activity. Of the monosaccharides tested for inhibition of precipitation of ricin, p-nitrophenyl betaGal was the best; this compound was 1.3 fold better than its alpha-anomer. While methyl betaGal was twice as active as its alpha anomer, Gal and blood group B active disaccharides (Galalpha1-3Gal) were 2.5 times more active than GalNAc. Among the oligosaccharides tested, Galbeta1-3GalNAc (T) Gal beta1-3/4GlcNAc (I/II), Galbeta1-4Glc (L) and human blood group I Ma trisaccharide (Galbeta1-4GlcNAcbeta1-6Gal) were about equally active and the best inhibitors. They were about 2.0 and 2.4 more active than Galalpha1-4Gal (E) sequence and B determinant, respectively. CONCLUSION: From the present results, it is concluded that: (a) this toxin has a broad range of affinity for the beta-anomer of Gal; (b) its combining site is probably of a shallow groove type and as large as a trisaccharide; (c) Galbeta--is the major combining site of the lectin; and (d) hydrophobic interaction gives a significant contribution for binding. This information should facilitate future usage of this lectin in glycobiological research and medical applications. PMID- 16265845 TI - Features of developmental functions and autistic profiles in children with fragile X syndrome. AB - BACKGROUND: In this study, we investigated the developmental functions and autistic profiles in children with Fragile X syndrome (FXS). In addition, we established the relationships between developmental and autistic profiles in these children. METHODS: The medical records of 12 children with FXS, aged 2 to 7 years, were collected. Fifteen children with autism, without FXS, who were age- and sex-matched were selected as the comparison group. All children underwent assessments of developmental functions according to the Chinese Child Development Inventory (CCDI), and autistic profiles according to the Childhood Autism Rating Scale (CARS). Differences in genders between the two groups were determined with the Fisher's exact test. Differences in developmental functions (CCDI) and autistic profiles (CARS) between the two groups were compared using Mann-Whitney U test and Bonferroni adjustment. The Spearman's rho correlation was used to determine the relationship of developmental functions and autistic profiles. RESULTS: All developmental functions in children with FXS were better than those with autism except for gross motor, fine motor and expressive language functions. Children with FXS had the worst expressive language function (56% Development Quotient, DQ) as compared with other developmental functions (> 70% DQ). The major difference between the children with FXS and those with autism was personal social function with a difference of 33% DQ. The average total CARS score were lower in children with FXS (average score, 28) than children with autism (average score, 34). Spearman's correlation demonstrated the CARS total score were negatively correlated with all developmental functions, except for gross motor function. CONCLUSIONS: Our findings suggest that the FXS children had multifaceted and disproportional development patterns in motor, speech and social domains when compared with the autism children without FXS. The developmental functions were inversely correlated with autistic profiles. Therefore, when applying comprehensive assessment, we were able to identify the special developmental features in children with FXS. PMID- 16265844 TI - Aggressive multimodality treatment for intra-hepatic recurrence of hepatocellular carcinoma following hepatic resection. AB - BACKGROUND: Intra-hepatic recurrence following hepatic resection is the primary challenge for patients with hepatocellular carcinoma (HCC). The aim of this investigation was to evaluate the long-term results of multimodality treatment of patients with intra-hepatic recurrent HCC following hepatic resection. METHODS: From January 1995 through December 2001, a total of 846 patients who underwent hepatic resection for primary HCC at the Chang Gung Memorial Hospital (Taoyuan, Taiwan) were analyzed through long-term follow-up; intra-hepatic recurrence of HCC was identified in 444 (52.5%) patients. Patients with intra-hepatic recurrence were categorized into two groups based on whether they underwent regional treatment. A comparison analysis of the survival rates of these two groups following treatment for recurrent intra-hepatic HCC was performed. RESULTS: Patients treated with multiple modalities exhibited significantly better survival results, both after initial hepatectomy and intra-hepatic recurrence compared with patients in the non-regional-treatment group. (p < 0.001) The 1-, 3 , and 5-year survival rates of patients in the regional-treatment group after initial hepatic resection were 88.1%, 60.1%, and 35.8%, respectively. The 1-, 3-, 5-year survival rates after treatment of intra-hepatic recurrence in the regional treatment group were 74.8%, 39.3%, and 25.2%, respectively. CONCLUSION: The incidence of intra-hepatic recurrence following hepatic resection for HCC remains high. Aggressive multimodality treatment could extend the survival for patients with recurrent postoperative intra-hepatic HCC. PMID- 16265847 TI - The effects of continuous axillary brachial plexus block with ropivacaine infusion on skin temperature and survival of crushed fingers after microsurgical replantation. AB - BACKGROUND: Continuous axillary brachial plexus block with local anesthetic has been shown to improve tissue perfusion after replantation surgery of the extremity. The present study aimed to investigate whether continuous axillary brachial plexus block with ropivacaine infusion can improve the survival of the reconstructive fingers secondary to an increase in its skin temperature in patients receiving replantation surgery of the crushed fingers. METHODS: Under general anesthesia, 18 patients received replantation or toe-to-hand transplantation of their crushed digits. They were randomly divided into two groups. Under ultrasound guidance, continuous axillary brachial plexus analgesia was effected by a loading dose of 10 ml 0.75% ropivacaine, followed by an infusion of 4-5 ml per hour for up to three days (Group A). Patients who did not receive continuous analgesia postoperatively served as a control (Group B). An infrared thermometer was used to hourly assess the skin temperature of the surgical and non-surgical sites in both groups for 24 h after the surgery. In addition, the survival (the rate of re-operation or amputation) of the reconstructive digits was also evaluated in both groups. RESULTS: The skin temperature of the digits (T1) on both groups did not show any significant difference at any point of time after the surgery albeit there was a trend of increased skin temperature on the reconstructive digits in patients receiving continuous axillary brachial plexus block (Group A) as compared to those without receiving the block (Group B). Also, the difference in skin temperature (dT) differed slightly at 0, 9 and 21 hours postoperatively in Group A in comparison with Group B (0.75 +/- 0.65 vs. -2.33 +/- 1.24, 0.53 +/- 0.34 vs. -3.02 +/- 1.27, -0.125 +/- 0.55 vs. -2.33 +/- 0.91, p < 0.05). However, no patients in both groups received a second operation or amputation of the graft. CONCLUSIONS: The result of this study demonstrated that axillary brachial plexus block with continuous infusion of 0.75% ropivacaine can increase the skin temperature, an index of tissue perfusion, of the reconstructive digits for 24 h after microvascular surgery of the crushed fingers. However, graft survival was good in both groups. PMID- 16265846 TI - Informational needs, health locus of control and uncertainty among women hospitalized with gynecological diseases. AB - BACKGROUND: Only a few studies on perceptions of uncertainty in illness have provided empirical evidence for the relationships of informational needs and the health locus of control with uncertainty among hospitalized women with gynecological diseases. The purpose of this study was to test Mishel's Theory of Uncertainty in Illness (1990) among women hospitalized with gynecological diseases. METHODS: Taiwan. The convenience sample consisted of 81 hospitalized women with gynecological diseases, who were invited to complete a set of self administered questionnaires prior to receiving any treatment. Path analysis was used to determine the relationships of informational needs and the health locus of control with uncertainty. RESULTS: The study findings suggested that hospitalized women's information needs are substantial, and they reported lower levels of uncertainty during hospitalization. Women's experience of uncertainty may be elevated by decreasing the informational needs as moderated by the beliefs that their health outcomes are under the control of chance. Three predictors in this model, i.e., informational needs moderated by the interaction of chance control, years of education, and number of treatments explained 30% of the variance of uncertainty of hospitalized women with gynecological diseases. CONCLUSIONS: The study findings suggest that healthcare professionals should carefully assess uncertainty levels among female patients with lower education and who believed that their health status depends upon external forces such as fate, luck, or chance. PMID- 16265848 TI - Chorioretinal folds associated with a meningioma. AB - Chorioretinal folds are a rare presentation of a meningioma. We report a case of a meningioma with chorioretinal folds as its initial presentation. A 39-year-old female patient complained of progressive blurring of the right eye. A fundus examination of the right eye revealed chorioretinal folds. Magnetic resonance imaging found a tumor mass involving the right orbit and right anterior middle cranial fossa. An echo-guided biopsy confirmed the meningioma. She received surgical excision of the meningioma, and her visual acuity was stationary at 12/20 after 15 months of follow-up. We review the etiologies of chorioretinal folds and elucidate the strategies for dealing with them. Certain underling diseases associated with chorioretinal folds are lethal and require prompt management. Ophthalmologists should evaluate these patients carefully in a stepwise manner. PMID- 16265849 TI - Serial follow-up in traumatic optic neuropathy using scanning laser polarimetry and visual field testing. AB - A 40-year-old male patient suffered from traumatic optic neuropathy in his right eye. Scanning laser polarimetry was arranged at 2 weeks, 9 weeks, 13 weeks, 24 weeks, and 34 weeks after the trauma. Manual or automated visual field testing was also arranged at 1 week, 5 weeks, 16 weeks, and 28 weeks correspondingly. The data revealed using scanning laser polarimetry (program GDx, version 1.0.05; Laser Diagnostic Technologies, San Diego, Calif, USA) were nearly normal at 2 weeks after trauma, but lower visual field loss was revealed using visual field testing within 2 weeks after the trauma. The superior hump of the GDx deviated from normal at about 9 weeks and some GDx parameters (the Number, Superior/Nasal, Ellipse Modulation (Ellipse Mod.), Maximal Modulation (Max. Mod.), Symmetry, Superior Ratio (Super. Ratio)) became worse later in the series. We propose that visual field defects might be present before retinal nerve fiber layer loss. In this case, scanning laser polarimetry for evaluating the severity of traumatic optic neuropathy was limited especially within 2 weeks after the trauma. PMID- 16265851 TI - The patient at the consulting room door. PMID- 16265850 TI - Meigs' syndrome in a young woman with a normal serum CA-125 level. AB - We report on a 27-year-old woman who presented with an ovarian solid tumor (20 x 15 cm) and massive ascites. A physical examination and chest X-ray revealed a moderate amount of pleural effusion on the right side. Cytologic study of the pleural effusion showed reactive mesothelial cells without evidence of malignancy. Gram's stain was negative. The blood chemistry was within normal limits. The serum CA-125 level was 22 (normal, < 35) U/ml, the alpha-fetoprotein (AFP) level was 8 (normal, < 20) ng/ml, and the carcinoembryonic antigen (CEA) was 0.5 (normal, < 5) ng/ml. An explorative laparotomy revealed approximately 1500 ml of serous ascites and a very large multilobulated left adnexal mass (20 x 15 cm) with no malignant cytology in the ascitic fluid. Postoperatively, the pleural effusion spontaneously resolved, and the microscopic examination revealed a benign fibroma-thecoma, confirming the diagnosis of Meigs' syndrome. The symptoms resolved after removal of this pelvic tumor. This is an unusual case of a young female with Meigs' syndrome and a normal serum CA-125 level. PMID- 16265852 TI - Bone response to submerged, unloaded implants inserted in poor bone sites: a histological and histomorphometrical study of 8 titanium implants retrieved from man. AB - An important parameter that influences the long-term success of oral implants is the bone quality of the implant bed. Posterior areas of the jaws have been avoided in implant dentistry because of their poor bone quality, higher chewing forces, and presumed higher implant failure rates. Several researchers have deemed soft bone implant sites to be a great potential risk situation, and most failures have been found in sites where the bone density was already low. The inferior success rates in the posterior maxilla have been attributed to a lower bone density and a lesser bone-implant interface. The aim of the present study was a histological and histomorphometrical analysis of the bone response to submerged implants inserted in posterior areas of the human jaws and retrieved, for different causes, after healing periods varying from 6 weeks to 12 months. Eight submerged implants that had been retrieved for different causes after different healing periods were evaluated in the present study. All implants were submerged and unloaded. Three implants had been removed for inadequate patient adaptation, 2 for inability of the implant to meet changed prosthetic needs, 1 for not optimal position from esthetic and hygiene aspects, and the last 2 for pain and dysesthesia. All the implants were retrieved with a 5-mm trephine bur. Newly formed peri-implant bone was found in all implants even after shorter healing periods. The bone-implant contact percentage varied from 30% to 96%. In conclusion, some surfaces have an improved characteristic of contact osteogenesis in soft bone, with coverage of the implant surface with a bone layer as a base for intensive bone formation and remodeling. We documented osseointegration of implants with a rough surface even after an insertion period of less than 2 months, both in the mandible and in the maxilla. From these results, we tentatively extrapolate that these implants might be carefully loaded after 2 months of healing, even when inserted in soft bone. A higher removal torque value might lead to a more predictable use of shorter implants, to a support of a prosthesis with fewer implants, or to shorter healing periods. PMID- 16265853 TI - Assessment of three-dimensional X-ray images: reconstruction from conventional tomograms, compact computerized tomography images, and multislice helical computerized tomography images. AB - Three-dimensional X-ray images (3D images) were used for imaging diagnosis in the oral and maxillofacial region. These images could be fundamentally reconstructed from various tomograms, though clinical 3D images were mainly reconstructed from computerized tomography (CT) images. In this investigation, 3D images were reconstructed from conventional tomograms with a panoramic unit, compact CT images, and multislice helical CT images, and the usefulness of each system was subjectively assessed for dental implant treatment. Three hemilateral dried human mandibles were used and were examined by linear tomography with a panoramic unit, compact CT, and multislice helical CT, and 3D images were reconstructed by using the rendering software for each system. The 3D images were visually evaluated on a 5-point scale covering the alveolar ridge, buccal and lingual bone surface, mental foramen, and tooth sockets. As a result, 3D images reconstructed from conventional tomograms with the panoramic unit were assessed as fair to unsure, compact CT 3D images were assessed as unsure to good, and multislice helical CT 3D images were assessed as good to excellent. It was concluded that compact CT 3D images and multislice helical CT 3D images were useful in dental implant treatment. PMID- 16265854 TI - Screw- vs cement-implant-retained restorations: an experimental study in the Beagle. Part 1. Screw and abutment loosening. AB - The causes of implant failures can be biological or mechanical. The mechanical causes include fracture of the implant, fracture of the abutment, and loosening of the abutment. Numerous studies show that abutment loosening constitutes one of the marked implant postsurgery complications requiring clinical intervention. The aim of the present study was to evaluate the incidence of the screw loosening in screwed or cemented abutments. Six adult male Beagles were used. In each dog, the first molars and 2 premolars were extracted. The sutures were removed after 7 days. After 3 months, 10 implants were placed in each dog, 5 in the right mandible and 5 in the left mandible. The abutments either were screwed in (n=30) by applying a total strength of 30 N/cm or were cemented (n=30). After 12 months, 8 (27%) loosened screws were present in screwed abutments, whereas no abutment loosening was observed in cemented abutments (P = .0001). Screwed abutments are often submitted to nonaxial loads that determine screw and abutment loosening. PMID- 16265855 TI - Histologic analysis of an immediately loaded implant retrieved after 2 months. AB - Human biopsy of immediately loaded implants is the most important way to determine the occurrence of osseointegration. Implants inserted in sites with poor bone quality have been associated with lower success rates. The aim of this study is to document the early healing processes in a man around an immediately loaded implant retrieved after a 2-month loading period. An implant was inserted in the mandible of a 32-year-old patient and was loaded into a nonfunctional loading mode with a fixed provisional prosthesis the same day of the implant surgery. After 2 months, because the patient had difficulty accepting the implant, the implant was retrieved with a 5-mm trephine drill. Before retrieval, the implant appeared to be clinically osseointegrated, and no mobility was present. The preexisting bone quality was type D4. The implant was surrounded by newly formed bone lamellae with a width of 200 to 400 microm. In many areas it was possible to observe osteoblasts producing osteoid matrix directly on the implant surface. Bone-to-implant contact percentage was 71% +/- 3.2%. Even in a poor bone site and after a healing period of only 2 months, we observed a high bone-to-implant contact percentage. We can confirm that immediately loaded implants placed in soft spongy bone after a 2-month healing period can present mineralized tissue at the interface. PMID- 16265856 TI - The use of prefabricated titanium tissue abutments for the construction of a maxillary subperiosteal implant. AB - This article presents a bimodal procedure for a patient with an implant-supported fixed partial denture (FP2 class of Misch) where endosteal implants (EOIs) and a subperiosteal implant (SPI) were indicated in different sections of the same arch. In edentulous patients, heterogeneous bone volume and density may be encountered in different regions of the same arch. When the available bone is favorable, the use of EOIs is simpler and less time consuming. An SPI is indicated when the available bone is severely resorbed, as in the type I division C, type II divisions C and D, and type III divisions C and D. The main advantage of the described combined technique is that it offers an alternative to invasive surgeries such as iliac crest bone grafts or various surgical augmentation procedures. The second advantage is the very brief healing period after insertion of the SPI before prosthetic loading. Planning, design, laboratory construction, surgical application, and prosthetic rehabilitation for bimodal implant treatment are described. Prefabricated titanium tissue abutments are used in the fabrication of the posts of the anterior titanium SPI. Titanium tissue abutments are placed in the waxup stage of the SPI before the casting of the titanium SPI. Posterior fixtures are EOIs. An anterior SPI with 4 posts and 4 posterior EOIs provides a combination of support for the screw-retained fixed maxillary denture of Misch's FP2. PMID- 16265857 TI - Bacterial infections in patients with cirrhosis. AB - Bacterial infection is a frequent and severe complication of cirrhosis that may present on admission or develop during hospitalization in 30-60% of hospitalized cirrhotic patients. The most frequent infective complications include spontaneous bacterial peritonitis, urinary tract infections, respiratory infections, and bacteremia, mostly due to the concomitant presence of various facilitating mechanisms such as changes in the reticuloendothelial system, decreased opsonic activity of the ascitic fluid, neutrophil leukocyte dysfunction, and iatrogenic factors. In fact, up to 25% of cases of death in cirrhotic patients are believed to be related to bacterial infections. This paper aims to provide a brief overview of the epidemiology, pathogenesis, treatment and prophylaxis of bacterial infection in cirrhosis. PMID- 16265858 TI - Interrelationships of maternal serum leptin, body mass index and gestational age. AB - BACKGROUND: The aim of this study was to evaluate the interrelationships between maternal serum leptin level and body mass index (BMI) at different gestational ages or during the whole pregnancy. METHODS: A total of 374 blood samples were obtained from 114 pregnant women when they visited the prenatal clinic for registration, Down syndrome screening test, 50 g oral glucose challenge test, hepatitis B markers test, and near or at the time of delivery. Serum leptin levels were measured by immunoradiometric assay. Linear regression analysis, 1 way ANOVA and post hoc tests were used to analyze the data. RESULTS: Throughout the course of pregnancy, maternal serum leptin concentration was significantly correlated with gestational age (p = 0.001, r = 0.179). A good correlation was also found between gestational age and maternal serum leptin level in the second trimester (p = 0.021, r = 0.158). A significant decline in maternal serum leptin was found in the third trimester (p = 0.011, r = 0.237). There were good correlations between maternal leptin and BMI in all 3 trimesters (p = 0.002 in the first trimester, p < 0.001 in the second trimester, p = 0.007 in the third trimester) and through the whole pregnancy (p < 0.001). Maternal BMI was related to gestational age in the second trimester (p < 0.001) and the whole pregnancy (p < 0.001), but not in the first (p = 0.404) and third trimesters (p = 0.053). CONCLUSION: Maternal serum leptin concentration was significantly related to gestational age (except in the first trimester) and BMI in the 3 trimesters and throughout pregnancy. Serum leptin concentration peaked during the early third trimester and declined significantly thereafter. Maternal BMI was related to gestational age in the second trimester and the whole pregnancy. PMID- 16265859 TI - Cholesteatoma surgery in pneumatized and non-pneumatized temporal bones. AB - BACKGROUND: This report compares the different surgical methods for large and pneumatized mastoid cavity (large cavity) and small and poorly pneumatized mastoid cavity (small cavity) cholesteatomas and their results for the facial nerve and mastoid cavity. METHODS: In the period from January 1996 to December 2000, 101 ears with cholesteatomas underwent tympanoplasty with mastoidectomy. In the large cavity group, the posterior canal wall was removed and the vertical portion of the facial nerve was skeletonized to lower the facial ridge. Partial obliteration with conchal cartilage and perichondrium at the epitympanum and mastoid tip was used to make the external auditory canal and the mastoid a smooth common cavity without reconstruction of the posterior canal wall. In the small cavity group, the facial ridge was not intentionally lowered as long as the cholesteatoma was completely removed. The whole mastoid cavity was obliterated and the posterior canal wall was reconstructed. The newly reconstructed posterior canal wall was composed of preserved facial ridge in the lower part and obliterated conchal cartilage in the upper part. RESULTS: There were 27 ears with large cavity cholesteatomas and 74 with small cavity cholesteatomas. The incidences of postoperative complications (large cavity versus small cavity) were: facial paralysis, 3 versus 0; minor wound infection, 2 versus 8; flap partial necrosis, 1 versus 4; cartilage partial necrosis, 0 versus 2; external auditory canal stenosis, 0 versus 2; residual drum perforation, 3 versus 3; otitis media with effusion, 3 versus 4; recurrent cholesteatoma, 0 versus 2; and residual cholesteatoma, 1 versus 2. CONCLUSION: Significantly more postoperative facial nerve paralysis in the large cavity group was correlated to skeletonization of the facial nerve and treatment of the facial ridge. PMID- 16265860 TI - Preliminary experience with botulinum toxin type A intracutaneous injection for Frey's syndrome. AB - BACKGROUND: Gustatory sweating, the main symptom of Frey's syndrome, usually occurs after parotid gland surgery. Numerous medical and surgical treatments have been proposed to treat this condition, but there are various drawbacks. Botulinum toxin type A intracutaneous injection is a relatively new treatment modality; its use has never been reported in Taiwan. Here, we present our preliminary experience with this technique and review the literature. METHODS: Between March 2002 and June 2005, 10 consecutive patients with severe gustatory sweating were managed with intracutaneous injection of botulinum toxin type A for a total of 16 times. The affected skin was visualized by Minor's starch-iodine test and then recorded. The interinjection distance was 1 cm and a mean dose of 46.4 U (at a concentration of 2.5 U/0.1 mL) was used. RESULTS: In all 10 cases, gustatory sweating improved within 2 days after injection, with no side effects. Patients in whom the first 13 injections were performed experienced recurrence of gustatory sweating. Mean duration of effectiveness was 9.3 months; the shortest duration of effectiveness was 2 months and the longest was 28 months. One patient also had gustatory flushing, but this symptom did not improve even after 3 treatments. CONCLUSION: Intracutaneous injection of botulinum toxin type A is a highly reliable, effective, safe, and minimally invasive treatment for gustatory sweating. Some patients had long-lasting therapeutic results. We recommend it as a valuable treatment option for severe cases of gustatory sweating. However, in our experience, it had no effect on facial skin flushing. Therefore, in addition to acetylcholine, there might be other neurotransmitters that are responsible for skin vasodilatation. PMID- 16265861 TI - Differential gene expression after hemorrhagic shock in rat lung. AB - BACKGROUND: We investigated the differential gene expression in rat lung after hemorrhagic shock (HS). METHODS: A controlled HS model in rats was used. Male Sprague-Dawley rats were randomly segregated into 2 groups, sham and HS. Samples of lung were procured from rats 2 hours after HS and resuscitation. Commercially available gene chips for rat cDNA microarray and software packages were used for the gene expression profile study. RESULTS: Compared with sham-shock rats, 98 genes were upregulated in HS rat lung. Most upregulated genes were responsible for inflammation (pro-inflammatory or anti-inflammatory cytokines, cognate receptors, and signal transduction for inflammation), protein activation (kinase and phosphatase), oxidation (oxidative and antioxidative enzymes), and apoptosis (apoptosis and anti-apoptosis). Eleven genes were downregulated after HS. CONCLUSION: HS may induce upregulation of positive and negative control genes responsible for inflammation, oxidation, protein metabolism and apoptosis, that is, a vulnerable period may develop in the host after HS. Overwhelming inflammatory response or immunosuppression may occur once a second hit, such as infection, ensues. Understanding, on a genome scale, how an organism responds to HS may facilitate the development of enhanced treatment modalities for HS. PMID- 16265862 TI - Operative treatment of intra-articular distal radius fractures using the small AO external fixation device. AB - BACKGROUND: A retrospective group study was done to evaluate the effect of the small AO external fixator in the management of acute intra-articular fractures of the distal radius. METHODS: Between January 1995 and December 1996, 70 consecutive patients with articular fractures of the distal radius were treated by closed reduction and external fixation with small AO external fixators. The mean age at the time of surgery was 58.9 years (range, 14-87 years). There were 58 Colles' Barton's fractures and 12 Smith's Barton's fractures. The follow-up period was 104 months (range, 92-118 months). RESULTS: All fractures united in a mean of 5.8 weeks (range, 4-10 weeks). At the final follow-up, the average range of motion was 56.3 +/- 11.6 degrees in flexion, 58.6 +/- 10.7 degrees in extension, 21.5 +/- 4.2 degrees in ulnar deviation, 9.1 +/- 2.9 degrees in radial deviation, 71.5 +/- 8.5 degrees in pronation, and 67.3 +/- 9.2 degrees in supination. Compared with the normal side, the average grip force was 87 +/- 6%. The overall clinical and functional outcomes, according to the scoring system of Gartland and Werley, showed that 22 patients (31.4%) had excellent results, 36 (51.4%) had good results, 9 (12.9%) had fair results, and 3 (4.3%) had poor results. CONCLUSION: Closed reduction and external fixation with the small AO external fixator is useful and effective in the management of displaced comminuted articular fractures of the distal radius. PMID- 16265863 TI - Radiofrequency catheter ablation of coexistent idiopathic left ventricular tachycardia and atrioventricular nodal reentrant tachycardia. AB - A healthy 15-year-old male patient presented with a 6-month history of recurrent attacks of palpitations. On multiple emergency room visits, a sustained wide QRS complex tachycardia with a right bundle branch block and northwest axis deviation was documented. The tachycardia was not terminated by intravenous adenosine, but was suppressed with intravenous verapamil. There was no evidence of structural heart disease, myocarditis, long QT syndrome, or electrolyte imbalance after a series of standard examinations. Idiopathic left ventricular tachycardia (ILVT) was suspected. Electrophysiologic studies revealed 2 inducible tachycardias, which were shown to represent atrioventricular nodal reentrant tachycardia (AVNRT) and ILVT. Transformation from AVNRT to ILVT occurred spontaneously following atrial pacing. Successful ablation of ILVT and the slow atrioventricular nodal pathway resulted in cure of the double tachycardia. PMID- 16265864 TI - Congenital diaphragmatic hernia with familial occurrence in a Taiwanese pedigree. AB - Congenital diaphragmatic hernia (CDH) is a developmental defect that accounts for 8% of all major congenital anomalies and is associated with a high mortality rate despite optimal postnatal treatment. Its etiology is uncertain. We report a case of familial CDH in a Taiwanese family. We believe autosomal recessive inheritance is the possible genetic etiology of CDH in this family. PMID- 16265865 TI - Spontaneous thrombosis and complete disappearance of traumatic carotid-cavernous fistulas after angiography. AB - Traumatic carotid-cavernous fistula (TCCF) is a direct shunting fistula due to a tear in the cavernous portion of the internal carotid artery (ICA). Spontaneous thrombosis of the high-flow shunts in TCCFs is extremely rare. Most cases are treated using endovascular embolization to relieve the clinical presentations. We report 2 unusual cases of TCCF with spontaneous closure of fistulas at intervals of 2 and 10 days, respectively, after diagnostic angiograms. The possible mechanisms of spontaneous healing of the fistulas in these patients with minor head injury and small fistulas were presumed to be a transient decrease in fistula blood flow because of irritation of and/or subtle injury to the ICA by contrast media and/or catheter during diagnostic angiogram, with thrombosis formation at the fistula. PMID- 16265866 TI - Delayed tension pneumothorax during surgery. AB - Tension pneumothorax is a life-threatening emergency that rapidly results in cardiopulmonary arrest. It requires prompt diagnosis and treatment. We present 2 cases from our practice, 1 caused by blunt chest trauma and the other resulting from laparoscopic surgery. Both were successfully treated by insertion of a chest tube. The diagnosis and treatment of intraoperative pneumothorax is discussed together with a review of the literature. PMID- 16265867 TI - [New results in the management of hypercholesterolemia]. AB - Cardiovascular disease is a major cause of morbidity and mortality worldwide. During the last decade huge amount of laboratory and clinical evidence proved link between serum lipid concentration and the development of coronary heart disease. The Adult Treatment Panel III of the National Cholesterol Education Program issued an evidence-based set of guidelines on cholesterol management. Since the publication of these guidelines numbers of new clinical trials and experimental results have been published. Recently completed clinical trials have indicated that reduction of low-density lipoprotein cholesterol to goals lower than the previously considered appropriate 2,6 mmol/l, produce further cardiovascular risk reduction in high risk individuals. There is growing evidence of the role of inflammation in the development of atherosclerosis and cardiovascular disease. Numerous studies have demonstrated that elevated C reactive protein may be an independent cardiovascular risk factor. Statin therapy has produced a greater cardiovascular risk reduction in those patients whose C reactive protein level was high. Ezetimibe is a new intestinal cholesterol uptake inhibitor, which reduces LDL cholesterol in monotherapy or in combination with statins. Lately a variety of new antilipemic agents are being developed. The present review summarizes some of these new results and their effect on cholesterol lowering therapy. PMID- 16265868 TI - [Continuous quality improvement with the use of new, evidence based quality indicators in the primary health care: there is a real possibility to restrain the unnecessary raising of antibiotic resistance]. AB - In the course of antibiotic treatments of the most frequent respiratory infections a lot of greatly heterogenic medical decisions in the primary health care were issued resulting in the irrational, untargeted and polypragmatic use of antibiotics, causing the spread of antibiotic resistance and increasing its threat world-wide. On the base of quality explorer investigations it was suggested that this bad attitude would be fairly characterised with use of our's new quality indicators which are really able to measure the negligence of the most important professional proposals (as the negligence of evidence based anticausal targeting) by doctors. In the future we would have to decrease the proportion of the erroneous medical decision patterns by virtue of the following of evidence based prescriptions of antibiotics and it would be helped by the introducing of continuous quality improvement based on the use of the new quality indicators. To regenerate the professional quality level of anticausal treatments we have to raise the frequency of rational, evidence based medical decision patterns to the desirable level to attain the avoidance of unnecessary or too broad spectrumed and thereby too expensive antibiotic treatments with an eye to restrain the spread of antibiotic resistance and to decrease the unnecessary costs. Today we have to choose expressly to serve the future of mankind or instead of those to serve the growth of industrial profit, only. PMID- 16265869 TI - [The long QT syndrome from the bedside to molecular genetic laboratory. The history of the first described Hungarian family]. AB - The authors demonstrate the developments of molecular genetic methods on the example of the first described Hungarian family suffering from inherited long QT (Romano-Ward) syndrome. The family belonged to the LQT2 subgroup according to both the clinical picture and the results of different molecular genetic methods from linkage mapping to sequencing. The final result of sequencing showed a missense mutation, affecting codon 568 in exon 7 of the KCNH2 (HERG) gene, leading to a tryptophan-cysteine change in the amino acid chain. PMID- 16265870 TI - [25 years' history of Varadi-Papp syndrome (orofaciodigital syndrome VI]. AB - INTRODUCTION: In 1978 the authors studied a male gypsy child with a multiple malformation syndrome. In this gypsy colony further five cases were found with similar features. The characteristic syndrome was published in 1980. Subsequently, the syndrome has been quoted as Varadi-Papp syndrome. AIM: To present the 25-year follow-up of this multiple malformation syndrome. RESULTS: The most common features of 29 affected children with this syndrome consists of orofacial (facial dysmorphism, cleft lip and/or palate abnormality, lingual nodule or tumor of the tongue, buccoalveolar frenula, alveolar and dental abnormalities, strabismus), cerebral/cerebellar (deformation of the skull, semilobar holoprosencephaly and/or absence or dysgenesis of cerebellar vermis or corpus callosum or hypothalamus or pituitary gland), digital (metacarpal abnormalities with central polydactyly, reduplication of the big toes) and genital (cryptorchidism, micropenis) anomalies. The patients are growth-retarded and when survival occurs psychomotor retardation is present. Accumulation of consanguinity and because of the involvement of multiple siblings in these families supports the autosomal recessive inheritance. CONCLUSION: Fetal Varadi Papp syndrome using ultrasonography in the mid-trimester both in routine screening and detailed scanning can be detected, and termination of pregnancy can be offered to the parents. PMID- 16265871 TI - [Wireless capsule endoscopy and its role in gastroenterology: our results and a review of the literature]. AB - INTRODUCTION: Since its recent introduction, capsule endoscopy has revolutionized the diagnostics of diseases of the small bowel. The number of studies on the use of this method is constantly increasing. Along with this, our knowledge about the usability and diagnostic power of capsule endoscopy is growing and the indications for this technique are getting progressively more accurate. AIMS: To study the usability of capsule endoscopy in suspected Crohn's disease, in gastrointestinal bleeding of small bowel origin, and in undetermined abdominal complains. METHOD: Capsule endoscopy was performed in 20, previously examined, carefully selected patients. RESULTS: Positive findings were recorded in all patients with gastrointestinal bleeding and in 3/4 of patients with suspected Crohn's disease. However, in patients with indeterminate abdominal complains, the capsule endoscopy proved to be inefficient. CONCLUSION: In the study below, based on their own experience, the authors try to provide an overall picture of capsule endoscopy by reviewing contemporary medical literature. The usability of the capsule endoscopy in suspected Crohn's disease and gastrointestinal bleeding of small bowel origin is suggested by the results of the study. PMID- 16265872 TI - [Late metastases of abdominal cutaneous malignant melanoma in the small and large bowels]. AB - The authors reported the case of a 56 years old man, who was operated with abdominal cutaneous malignant melanoma 5 years ago. He had chemo-immunotherapy. His complaints were epigastric pain, melena, hematochezia, anorexia, lack of appetite, fatigue. The upper panendoscopy showed tumor mass in the duodenojejunal flexure and the colonoscopy showed tumor in the large bowel. The patient underwent jejunal resection and right hemicolectomy. The authors survey the metastases of malignant melanoma as well as their clinical signs, therapeutic measures and prognosis. PMID- 16265873 TI - [Treatment of urinary bladder cancers]. PMID- 16265874 TI - Dawn of a new era in molecular cancer therapeutics. PMID- 16265875 TI - Obstacles and opportunities in the clinical development of targeted therapeutics. PMID- 16265877 TI - Angiogenesis inhibitors: what is the clinical future? PMID- 16265876 TI - Tumor models for preclinical development of targeted agents. PMID- 16265878 TI - Epidermal growth factor receptor (EGFR) inhibitors in cancer therapy. PMID- 16265879 TI - Cell survival signaling during apoptosis: implications in drug resistance and anti-cancer therapeutic development. PMID- 16265880 TI - Targeted histone deacetylase inhibition for cancer prevention and therapy. PMID- 16265881 TI - Inhibitors of cyclin-dependent kinase modulators for cancer therapy. AB - Most human malignancies have an aberration in the Rb pathway due to 'cdk hyperactivation'. Several small-molecule cdk modulators are being discovered and tested in the clinic. The first ATP-competitive cdk inhibitors tested in clinical trials, flavopiridol and UCN-01, have shown promising results with evidence of antitumor activity and plasma concentrations sufficient to inhibit cdk-related functions. The best schedule to be administered, combination with standard chemotherapeutic agents, best tumor types to be targeted, and demonstration of cdk modulation from tumor samples from patients in these trials are important issues that need to be answered to advance these agents to the clinical arena. PMID- 16265882 TI - Targeting cyclooxygenase-2 for cancer prevention and treatment. PMID- 16265883 TI - Antisense approaches in drug discovery and development. PMID- 16265884 TI - Preclinical development of Alimta (Pemetrexed, LY231514), a multitargeted antifolate. PMID- 16265885 TI - Quinolines and artemisinin: chemistry, biology and history. AB - Plasmodium falciparum is the most important parasitic pathogen in humans, causing hundreds of millions of malaria infections and millions of deaths each year. At present there is no effective malaria vaccine and malaria therapy is totally reliant on the use of drugs. New drugs are urgently needed because of the rapid evolution and spread of parasite resistance to the current therapies. Drug resistance is one of the major factors contributing to the resurgence of malaria, especially resistance to the most affordable drugs such as chloroquine. We need to fully understand the antimalarial mode of action of the existing drugs and the way that the parasite becomes resistant to them in order to design and develop the new therapies that are so urgently needed. In respect of the quinolines and artemisinins, great progress has been made recently in studying the mechanisms of drug action and drug resistance in malaria parasites. Here we summarize from a historical, biological and chemical, perspective the exciting new advances that have been made in the study of these important antimalarial drugs. PMID- 16265886 TI - Antimalarial multi-drug resistance in Asia: mechanisms and assessment. AB - The emergence and spread of drug-resistant parasites poses a major problem for management of Plasmodium falciparum malaria in endemic areas. Nowhere is this more apparent than in southeast Asia, where multi-drug resistance to chloroquine and sulfadoxine-pyrimethamine was exacerbated when mefloquine monotherapy began failing in the 1980s. A better understanding of mechanisms of (multi-) drug resistance is urgently warranted to monitor and guide antimalarial chemotherapy regimens more efficiently. Here we review recent advances on identification of molecular markers that can be employed in predicting in vitro and in vivo resistance in southeast Asia. Examples include amplification of PfMDR1 (P. falciparum multi-drug resistant gene 1) and mefloquine, K76T PfCRT and chloroquine, as well as mutations in the dihydroperoate synthase and dihydrofolate reductase genes and the antifolate class of drugs. PMID- 16265887 TI - Antimalarial drug resistance in Africa: strategies for monitoring and deterrence. AB - Despite the initiation in 1998 by the World Health Organization of a campaign to 'Roll Back Malaria', the rates of disease and death caused by Plasmodium falciparum malaria in sub-Saharan Africa are growing. Drug resistance has been implicated as one of the main factors in this disturbing trend. The efforts of international agencies, governments, public health officials, advocacy groups and researchers to devise effective strategies to deter the spread of drug resistant malaria and to ameliorate its heavy burden on the people of Africa have not succeeded. This review will not attempt to describe the regional distribution of drug resistant malaria in Africa in detail, mainly because information on resistance is limited and has been collected using different methods, making it difficult to interpret. Instead, the problems of defining and monitoring resistance and antimalarial drug treatment outcomes will be discussed in hopes of clarifying the issues and identifying ways to move forward in a more coordinated fashion. Strategies to improve measurement of resistance and treatment outcomes, collection and use of information on resistance, and potential approaches to deter and reduce the impact of resistance, will all be considered. The epidemiological setting and the goals of monitoring determine how antimalarial treatment responses should be measured. Longitudinal studies, with incidence of uncomplicated malaria episodes as the primary endpoint, provide the best information on which to base treatment policy changes, while simpler standard in vivo efficacy studies are better suited for ongoing efficacy monitoring. In the absence of an ideal antimalarial combination regimen, different treatment alternatives are appropriate in different settings. But where chloroquine has failed, policy changes are long overdue and action must be taken now. PMID- 16265888 TI - Uncomplicated malaria. AB - All symptoms and signs of uncomplicated malaria are non-specific, as shared with other febrile conditions, and can occur early or later in the course of the disease. In endemic areas, the presence of hepatosplenomegaly, thrombocytopenia and anaemia is clearly associated with malaria, particularly in children. Fever, cephalgias, fatigue, malaise, and musculoskeletal pain constitute the most frequent clinical features in malaria. Following single exposure to Plasmodium falciparum infection, the patient will either die in the acute attack or survive with the development of some immunity. Elderly individuals are prone to a more severe course of disease. The non-fatal P. vivax and P. ovale cause similar initial illnesses, with bouts of fever relapsing periodically, but irregularly over a period of up to 5 years. Renal involvement of a moderate degree is more common in mild falciparum malaria than initially suspected. The liver is also afflicted in mild disease, but organ damage is limited and fully reversible after parasitological cure. Whereas the cardiotoxic adverse effects of antimalarial chemotherapeutics are well known, clinically relevant cardiac involvement in humans is rare in severe disease and even rarer in uncomplicated falciparum malaria. Co-infection can aggravate malaria. There is a growing body of evidence that there is significant interaction in terms of mutual aggravation of the course of disease between HIV and malaria, particularly in pregnant women. Children with a high level of exposure to P. falciparum have a lower risk of developing atopic disorders. PMID- 16265889 TI - Metabolic complications of severe malaria. AB - Metabolic complications of malaria are increasingly recognized as contributing to severe and fatal malaria. Disorders of carbohydrate metabolism, including hypoglycaemia and lactic acidosis, are amongst the most important markers of disease severity both in adults and children infected with Plasmodium falciparum. Amino acid and lipid metabolism are also altered by malaria. In adults, hypoglycaemia is associated with increased glucose turnover and quinine-induced hyperinsulinaemia, which causes increased peripheral uptake of glucose. Hypoglycaemia in children results from a combination of decreased production and/or increased peripheral uptake of glucose, due to increased anaerobic glycolysis. Patients with severe malaria should be monitored frequently for hypoglycaemia and treated rapidly with intravenous glucose if hypoglycaemia is detected. The most common aetiology of hyperlactataemia in severe malaria is probably increased anaerobic glucose metabolism, caused by generalized microvascular sequestration of parasitized erythrocytes that reduces blood flow to tissues. Several potential treatments for hyperlactataemia have been investigated, but their effect on mortality from severe malaria has not been determined. PMID- 16265890 TI - The clinical and pathophysiological features of malarial anaemia. AB - This review will focus on the principal clinical and pathophysiological features of the anaemia of falciparum malaria, including the problems of treating malarial anaemia, and also will suggest how recent advances in genomics may help our understanding of cellular and molecular mechanisms underlying this syndrome. PMID- 16265891 TI - Malaria in the pregnant woman. AB - Women become more susceptible to Plasmodium falciparum malaria during pregnancy, and the risk of disease and death is high for both the mother and her fetus. In low transmission areas, women of all parities are at risk for severe syndromes like cerebral malaria, and maternal and fetal mortality are high. In high transmission areas, where women are most susceptible during their first pregnancies, severe syndromes like cerebral malaria are uncommon, but severe maternal anemia and low birth weight are frequent sequelae and account for an enormous loss of life. P. falciparum-infected red cells sequester in the intervillous space of the placenta, where they adhere to chondroitin sulfate A but not to receptors like CD36 that commonly support adhesion of parasites infecting nonpregnant hosts. Poor pregnancy outcomes due to malaria are related to the macrophage-rich infiltrates and pro-inflammatory cytokines such as tumor necrosis factor-alpha that accumulate in the intervillous space. Women who acquire antibodies against chrondroitin sulfate A (CSA)-binding parasites are less likely to have placental malaria, and are more likely to deliver healthy babies. In areas of stable transmission, women acquire antibodies against CSA binding parasites over successive pregnancies, explaining the high susceptibility to malaria during first pregnancy, and suggesting that a vaccine to prevent pregnancy malaria should target placental parasites. Prevention and treatment of malaria are essential components of antenatal care in endemic areas, but require special considerations during pregnancy. Recrudescence after drug treatment is more common during pregnancy, and the spread of drug-resistant parasites has eroded the usefulness of the few drugs known to be safe for the woman and her fetus. Determining the safety and effectiveness of newer antimalarials in pregnant women is an urgent priority. A vaccine that prevents pregnancy malaria due to P. falciparum could be delivered before first pregnancy, and would have an enormous impact on mother-child health in tropical areas. PMID- 16265892 TI - Host receptors in malaria merozoite invasion. AB - The clinical manifestations of Plasmodium falciparum malaria are directly linked to the blood stage of the parasite life cycle. At the blood stage, the circulating merozoites invade erythrocytes via a specific invasion pathway often identified with its dependence or independence on sialic acid residues of the host receptor. The invasion process involves multiple receptor-ligand interactions that mediate a complex series of events in a period of approximately 1 min. Although the mechanism by which merozoites invade erythrocytes is not fully understood, recent advances have put a new perspective on the importance of developing a multivalent blood stage-malaria vaccine. In this review, we highlight the role of currently identified host invasion receptors in blood-stage malaria. PMID- 16265893 TI - A post-genomic view of the mitochondrion in malaria parasites. AB - Mitochondria in Plasmodium parasites have many characteristics that distinguish them from mammalian mitochondria. Selective targeting of malaria parasite mitochondrial physiology has been exploited in successful antimalarial chemotherapy. At present, our understanding of the functions served by the parasite mitochondrion is somewhat limited, but the availability of the genomic sequences makes it possible to develop a framework of possible mitochondrial functions by providing information on genes encoding mitochondrially targeted proteins. This review aims to provide an overview of mitochondrial physiology in this post-genomic era. Although in many cases direct experimental proof for their mitochondrial functions may not be available at present, descriptions of these potential mitochondrial proteins can provide a basis for experimental approaches. PMID- 16265894 TI - The plastid of Plasmodium spp.: a target for inhibitors. AB - Determined efforts are being made to explore the non-photosynthetic plastid organelle of Plasmodium falciparum as a target for drug development. Certain antibiotics that block organellar protein synthesis are already in clinical use as antimalarials. However, all the indications are that these should be used only in combination with conventional antimalarials. The use of antibiotics such as doxycycline and clindamycin may reduce the development of drug resistant parasites and such means to avoid drug resistance should be explored hand-in-hand with drug development. Genomic information predicts that fatty acid type II (FAS II) and isoprenoid biosynthetic pathways are localized to the plastid. However, clinical trials with fosmidomycin (a specific inhibitor of DOXP reductase in the non-mevalonate pathway for isoprenoids) suggest it too should only be used in drug combinations. Prospects for more potent antimalarial compounds have emerged from studies of several of the enzymes involved in the FAS II pathway. Lead antibiotics such as thiolactomycin (an inhibitor of beta-ketoacyl-ACP synthase) and triclosan (a specific inhibitor of enoyl-ACP reductase) have led to structurally similar, active compounds that rapidly kill ring- and trophozoite stage parasites. The FAS II pathway is of particular interest to the pharma industry. PMID- 16265895 TI - Hemoglobin degradation. AB - Hemoglobin degradation by Plasmodium is a massive catabolic process within the parasite food vacuole that is important for the organism's survival in its host erythrocyte. A proteolytic pathway is responsible for generating amino acids from hemoglobin. Each of the enzymes involved has its own peculiarities to be exploited for development of antimalarial agents that will starve the parasite or result in build-up of toxic intermediates. There are a number of unanswered questions concerning the cell biology, biochemistry and metabolic roles of this crucial pathway. PMID- 16265896 TI - Bioavailable iron and heme metabolism in Plasmodium falciparum. AB - Iron metabolism is essential for cell function and potentially toxic because iron can catalyze oxygen radical production. Malaria-attributable anemia and iron deficiency anemia coincide as being treatable diseases in the developing world. In absolute amounts, more than 95% of Plasmodium metal biochemistry occurs in the acidic digestive vacuole where heme released from hemoglobin catabolism forms heme crystals. The antimalarial quinolines interfere with crystallization. Despite the completion of the Plasmodium genome, many 'gene gaps' exist in components of the metal pathways described in mammalian or yeast cells. Present evidence suggests that parasite bioavailable iron originates from a labile erythrocyte cytosolic pool rather than from abundant heme iron. Indeed the parasite has to make its own heme within two separate organelles, the mitochondrion and the apicomplast. Paradoxically, despite the abundance of iron within the erythrocyte, iron chelators are cytocidal to the Plasmodium parasite. Hemozoin has become a sensitive biomarker for laser desorption mass spectrometry detection of Plasmodium infection in both mice and humans. PMID- 16265897 TI - Plasmodium permeomics: membrane transport proteins in the malaria parasite. AB - Membrane transport proteins are integral membrane proteins that mediate the passage across the membrane bilayer of specific molecules and/or ions. Such proteins serve a diverse range of physiological roles, mediating the uptake of nutrients into cells, the removal of metabolic wastes and xenobiotics (including drugs), and the generation and maintenance of transmembrane electrochemical gradients. In this chapter we review the present state of knowledge of the membrane transport mechanisms underlying the cell physiology of the intraerythrocytic malaria parasite and its host cell, considering in particular physiological measurements on the parasite and parasitized erythrocyte, the annotation of transport proteins in the Plasmodium genome, and molecular methods used to analyze transport protein function. PMID- 16265899 TI - Molecular genetics of mosquito resistance to malaria parasites. AB - Malaria parasites are transmitted by the bite of an infected mosquito, but even efficient vector species possess multiple mechanisms that together destroy most of the parasites present in an infection. Variation between individual mosquitoes has allowed genetic analysis and mapping of loci controlling several resistance traits, and the underlying mechanisms of mosquito response to infection are being described using genomic tools such as transcriptional and proteomic analysis. Malaria infection imposes fitness costs on the vector, but various forms of resistance inflict their own costs, likely leading to an evolutionary tradeoff between infection and resistance. Plasmodium development can be successfully completed onlyin compatible mosquito-parasite species combinations, and resistance also appears to have parasite specificity. Studies of Drosophila, where genetic variation in immunocompetence is pervasive in wild populations, offer a comparative context for understanding coevolution of the mosquito-malaria relationship. More broadly, plants also possess systems of pathogen resistance with features that are structurally conserved in animal innate immunity, including insects, and genomic datasets now permit useful comparisons of resistance models even between such diverse organisms. PMID- 16265898 TI - Plasmodium ookinete invasion of the mosquito midgut. AB - The Plasmodium ookinete is the developmental stage of the malaria parasite that invades the mosquito midgut. The ookinete faces two physical barriers in the midgut which it must traverse to become an oocyst: the chitin- and protein containing peritrophic matrix; and the midgut epithelial cell. This chapter will consider basic aspects of ookinete biology, molecules known to be involved in midgut invasion, and cellular processes of the ookinete that facilitate parasite invasion. Detailed knowledge of these mechanisms may be exploitable in the future towards developing novel strategies of blocking malaria transmission. PMID- 16265900 TI - Functional proteome and expression analysis of sporozoites and hepatic stages of malaria development. AB - An evolution in modern malaria research occurred with the completion of the Plasmodium falciparum genome project and the onset and application of novel post genomic technologies. Corresponding with these technological achievements are improvements in accessing and purifying parasite material from 'hard-to-reach' stages of malaria development. Characterization of gene and protein expression in the infectious sporozoite and subsequent liver-stage parasite development is critical to identify novel pre-erythrocytic drug and vaccine targets as well as to understand the basic biology of this deadly parasite. Both transcriptional and proteomic analyses on these stages and the remaining stages of development will assist in the 'credentialing process' of the complete malaria genome. PMID- 16265901 TI - The dissection of CD8 T cells during liver-stage infection. AB - Multiple injections of gamma-radiation-attenuated Plasmodium sporozoites (gamma spz) can induce long-lived, sterile immunity against pre-erythrocytic stages of malaria. Malaria antigen (Ag)-specific CD8 T cells that produce IFN-gamma are key effector cells in this model of protection. Although there have been numerous reports dealing with gamma-spz-induced CD8 T cells in the spleen, CD8 T cells most likely confer protection by targeting infected hepatocytes. Consequently, in this chapter we discuss observations and hypotheses concerning CD8 T cell responses that occur in the liver after an encounter with the Plasmodium parasite. Protracted protection against pre-erythrocytic stages requires memory CD8 T cells and we discuss evidence that gamma-spz-induced immunity is indeed accompanied by the presence of intrahepatic CD44hi CD45RBlo CD62lo CD122lo effector memory (EM) CD8 T cells and CD44hi CD45RBhi CD621hi CD122hi central memory (CM) CD8 T cells. In addition, the EM CD8 T cells rapidly release IFN gamma in response to spz challenge. The possible role of Kupffer cells in the processing of spz Ags and the production of cytokines is also considered. Finally, we discuss evidence that is consistent with a model whereby intrahepatic CM CD8 T cells are maintained by IL-15 mediated-homeostatic proliferation while the EM CD8 T cells are conscripted from the CM pool in response to a persisting depot of liver-stage Ag. PMID- 16265902 TI - Early interactions between blood-stage plasmodium parasites and the immune system. AB - Accumulating evidence provides strong support for the importance of innate immunity in shaping the subsequent adaptive immune response to blood-stage Plasmodium parasites, the causative agents of malaria. Early interactions between blood-stage parasites and cells of the innate immune system, including dendritic cells, monocytes/macrophages, natural killer (NK) cells, NKT cells, and gamma6 T cells, are important in the timely control of parasite replication and in the subsequent elimination and resolution of the infection. The major role of innate immunity appears to be the production of immunoregulatory cytokines, such as interleukin (IL)-12 and interferon (IFN)-gamma, which are critical for the development of type 1 immune responses involving CD4+ Thl cells, B cells, and effector cells which mediate cell-mediated and antibody-dependent adaptive immune responses. In addition, it is likely that cells of the innate immune system, especially dendritic cells, serve as antigen-presenting cells. Here, we review recent data from rodent models of blood-stage malaria and from human studies, and outline the early interactions of infected red blood cells with the innate immune system. We compare and contrast the results derived from studies in infected laboratory mice and humans. These host species are sufficiently different with respect to the identity of the infecting Plasmodium species, the resulting pathologies, and immune responses, particularly where the innate immune response is concerned. The implications of these findings for the development of an effective and safe malaria vaccine are also discussed. PMID- 16265903 TI - Longevity of the immune response and memory to blood-stage malaria infection. AB - Immunity to malaria develops slowly with protection against the parasite lagging behind protection against disease symptoms. The data on the longevity of protective immune responses are sparse. However, studies of antibody responses associated with protection reveal that they consist of a short- and a long-lived component. Compared with the antibody levels observed in other infection and immunization systems, the levels of the short-lived antibody compartment drop below the detectable threshold with unusual rapidity. The prevalence of long lived antibodies is comparable to that seen after bacterial and protozoan infections. There is even less available data concerning T cell longevity in malaria infection, but what there is seems to indicate that T cell memory is short in the absence of persistent antigen. In general, the degree and duration of parasite persistence represent a major factor determining how immune response longevity and protection correlate. The predilection for short-lived immune responses in malaria infection could be caused by a number of mechanisms resulting from the interplay of normal regulatory mechanisms of the immune system and immune evasion by the parasite. In conclusion, it appears that the parasite host relationship has developed to favor some short-lived responses, which allow the host to survive while allowing the parasite to persist. Anti-malarial immune responses present a complex picture, and many aspects of regulation and longevity of the response require further research. PMID- 16265904 TI - Experimental models of cerebral malaria. AB - Malaria remains a major global health problem and cerebral malaria is one of the most serious complications of this disease. Recent years have seen important advances in our understanding of the pathogenesis of cerebral malaria. Extensive analysis of tissues and blood taken from patients with cerebral malaria has been complimented by the use of animal models to identify specific components of pathogenic pathways. In particular, an important role for CD8+ T cells has been uncovered, as well divergent roles for members of the tumor necrosis factor (TNF) family of molecules, including TNF and lymphotoxin alpha. It has become apparent that there maybe more than one pathogenic pathway leading to cerebral malaria. The last few years have also seen the testing of vaccines designed to target malaria molecules that stimulate inflammatory responses and thereby prevent the development of cerebral malaria. In this review, we will discuss the above advancements, as well as other important findings in research into the pathogenesis of cerebral malaria. As our understanding of pathogenic responses to Plasmodium parasites gathers momentum, the chance of a breakthrough in the development of treatments and vaccines to prevent death from cerebral malaria have become more realistic. PMID- 16265905 TI - Glycosylphosphatidylinositols in malaria pathogenesis and immunity: potential for therapeutic inhibition and vaccination. AB - Glycosylphosphatidylinositols (GPIs) are found in the outer cell membranes of all eukaryotes. GPIs anchor a diverse range of proteins to the surface of Plasmodium falciparum, but may also exist free of protein attachment. In vitro and in vivo studies have established GPIs as likely candidate toxins in malaria, consistent with the prevailing paradigm that attributes induction of inflammatory cytokines, fever and other pathology to parasite toxins released when schizonts rupture. Although evolutionarily conserved, sufficient structural differences appear to exist that impart upon plasmodial GPIs the ability to activate second messengers in mammalian cells and elicit immune responses. In populations exposed to P. falciparum, the antibody response to purified GPIs is characterised by a predominance of immunoglobulin (Ig)G over IgM and an increase in the prevalence, level and persistence of responses with increasing age. It remains unclear, however, if these antibodies or other cellular responses to GPIs mediate anti toxic immunity in humans; anti-toxic immunity may comprise either reduction in the severity of disease or maintenance of the malaria-tolerant state (i.e. persistent asymptomatic parasitaemia). P. falciparum GPIs are potentially amenable to specific therapeutic inhibition and vaccination; more needs to be known about their dual roles in malaria pathogenesis and protection for these strategies to succeed. PMID- 16265906 TI - The immunology and pathogenesis of malaria during pregnancy. AB - Women in endemic areas become highly susceptible to malaria during first and second pregnancies, despite immunity acquired after years of exposure. Recent insights have advanced our understanding of pregnancy malaria caused by Plasmodium falciparum, which is responsible for the bulk of severe disease and death. Accumulation of parasitized erythrocytes in the blood spaces of the placenta is a key feature of maternal infection with P. falciparum. Placental parasites express surface ligands and antigens that differ from those of other P. falciparum variants, facilitating evasion of existing immunity, and mediate adhesion to specific molecules, such as chondroitin sulfate A, in the placenta. The polymorphic and clonally variant P. falciparum erythrocyte membrane protein 1, encoded by var genes, binds to placental receptors in vitro and may be the target of protective antibodies. An intense infiltration of immune cells, including macrophages, into the placental intervillous spaces, and the production of pro-inflammatory cytokines often occur in response to infection, and are associated with low birth weight and maternal anemia. Expression of alpha and beta chemokines may initiate or facilitate this cellular infiltration during placental malaria. Specific immunity against placental-binding parasites may prevent infection or facilitate clearance of parasites prior to the influx of inflammatory cells, thereby avoiding a cascade of events leading to disease and death. Much less is known about pathogenic processes in P. vivax infections, and corresponding immune responses. Emerging knowledge of the pathogenesis and immunology of malaria in pregnancy will increasingly lead to new opportunities for the development of therapeutic and preventive interventions and new tools for diagnosis and monitoring. PMID- 16265907 TI - [The study of physico-chemical properties of iodinated proteins for iodine deficiency prophylaxis]. AB - Iodine is an essential trace element, its deficiency leads to iodine-deficient diseases. Utilization of iodinated milk proteins in human diet seems to be one of possible decisions of the problem. The aim of the present work was to characterize iodinated milk proteins "Bioiod", produced by the original technology of "SPF Technovita". The presence of iodinated amino acids (mono- and di-iodotyrosine) was proved by iodinated proteins hydrolyzates HPLC--MALDI mass spectrometry was used for the determination of iodinated proteins. The percentage of iodine incorporated into proteins was proved to be more then 2%. PMID- 16265908 TI - [Dietary intake of antioxidant vitamins in Russian population]. AB - Aim of present study was the getting data on dietary intake of antioxidant vitamins in Russian population in 1995-2002. The consumption of food was estimated by 24-hour recall methodology in course of Russian Longitudinal monitoring surveys of economics and health. Mean intake of vitamin A is less than recommended level and 25th procentile is near to zero. Mean and median of vitamin E intake are equal of recommended level. Dietary intake of vitamin A per 1000 kcal is maximal in group of children 0-6 year old and in adults aged 30-60 y. Nutrient density of daly diet in relation to antioxidant vitamins has maximal level in old group of population 60+ and minimal level in 6-18 year old children. PMID- 16265909 TI - [Revalence of vitamin deficiency in Tajik children]. AB - A research of estimating the prevalence og vitamin A deficiency in Tajik children was made. At revealed that 51.8% of children of age 6-59 months suffer from vitamin A defficiency. 97.4% of the examined pregnant women have no information at all about vitamin A and didnit take any vitamins including vitamin A. PMID- 16265910 TI - [Methodology of use of an enteral nutrition formulas in diet therapy of patients with type 2 diabetes]. AB - In this review the nutrient structure of standard and specialized enteral formulas for patients with diabetes mellitus is given and the data about medical efficacy specialized enteral formulas in correction of clinical and metabolic disorders in this contingent of patients are discussed. PMID- 16265911 TI - [Safety of food supplements containing plant components]. AB - For today the biologically active additives (BAA) extensively use by people as a source essential substances. However majoriry of people don't know, that BAA which contain vegetable components may induces various undiserable effects (UE). Review contain information about frequency of serious UE, first of all induced BAA, which contain medicinal plants. Discussed problem of charge efficacy and safety drugs when its use jointly with BAA. Think that need to give in BAA instruction additional information about possibility UE BAA. PMID- 16265912 TI - [The study of influence of chitosan on clinical, metabolic and immune parameters in patients with cardiovascular diseases]. AB - The investigation of influent of antiatherosclerotic diets with chitosan on clinical and metabolic parameters in patients with cardiovascular diseases. Results of the study show that enrichment of a diet with chitosan in patients with ishemic heart disease and hypertension improved clinic, immune status, antropometric levels and lipid spectrum of blood. The research has shown, that the use in the treat-preventive purposes chitosan is rather perspective. PMID- 16265913 TI - [Hypolipidemic activity of a water extract from roots carline thistle]. AB - In the researches which have been lead on white not purebred rats--mans on model nutritional hyperlipoproteinemia it fixed hypolipidemic activity of a water extract from roots carline thistle with the content of dry solvends of 7% which. PMID- 16265914 TI - [Impact of dietotherapy with enterosorbent of marine origin on the indices of mineral and lipid metabolism for patients suffering from kidney diseases]. AB - It is described the effectiveness of application of dietotherapy including enterosorbent of marine origin at the stage of rehabilitation of patients having kidneys pathology. Application of this enterosorbent favors the positive clinic biochemical dynamics with correction of electrolyte and lipid imbalance. PMID- 16265915 TI - [Clinical manifestations of cross-allergic reactions to food among children]. AB - Scientists have recently studied the frequency and possible forms of clinical manifestations of cross-reactivity to food among 239 of Abakan at the age from 6 month to 15 years with skin, respiratory and combined manifestation of atopy It was proved that risk factors of development of cross-reactivity to food occur among 80% of children who have various forms of allergic diseases moreover the cross allergic reactions between the foodstuffs most frequently occur with the gastrointestinal and with skin forms of food allergy and the cross allergic reactions between the pollen and food allergens more frequently occut with the respiratory forms of atopy and dermorespiratory syndrome. Considerably more rarely this type of reactions is discovered between the epidermal and food allergens, which most frequently were revealed with the dermatorespiratory syndrome. PMID- 16265916 TI - [Vitamin metabolism and status in patients with gastrointestinal diseases]. AB - Principle differences in vitamin C, B2, B6, A, E and carotenoids sufficiency of healthy suffering from gastrointestinal diseases (gastric and duodenal ulcer, chronic gastritis, dyskinesia bile tracts) ulc adults and children has not been determined by means of the statistic analysis including such parameters as median, mode, insufficiency frequency, curves of vitamin blood plasma concentration distribution. Polyhypovitaminosis frequency among patients is a result of low vitamin content in reduced diets but not reflect vitamin metabolism alteration. PMID- 16265917 TI - [Utilize of mathematical modeling methods for human health risk assessment from exposure to food contaminants]. AB - The present review reports on the mathematical methods and statistical techniques available for human health risk assessment from exposure to food contaminants. A framework is established for the usage of these methods instead of traditional toxicological techniques. Regarding to each method applicability, strengths, limitations and weaknesses are described. PMID- 16265918 TI - Bicycle injury prevention and safety in senior riders. AB - Senior bicycle riders have special safety and injury prevention needs. They are at greater risk for injury and mortality. Very few educational materials have been developed to assist this group in improving their riding abilities and preventing injury to themselves. This article reviews the risks encountered by older bicyclists, discusses injury revention, and describes the development of an educational video developed especially for senior riders consistent with the goals of the National Bicycle Safety Conference. PMID- 16265919 TI - Traumatic injury to the pancreas: the challenges of care in the pediatric patient. AB - Traumatic pancreatic injury is infrequently encountered in children. Diagnosis and treatment of this injury can be complicated. Signs and symptoms are often subtle with presentation frequently delayed leading to increased morbidity and mortality. This delay is compounded by lack of specific diagnostic tools to aid diagnosis. Clinicians should maintain a high index of suspicion for pancreatic injury in any child who sustains blunt abdominal trauma. Ongoing clinical evaluation is essential. This article presents a case study of a child with a traumatic pancreatic injury and discusses tools utilized to aid diagnosis of pancreatic injury, treatment options, and potential complications. PMID- 16265920 TI - Video recording of emergency department trauma resuscitations. AB - Although hospitals are faced with the challenges of appropriately informing the public regarding health care and protecting the privacy of patients, a comprehensive policy concerning videotaping of trauma resuscitations can be developed to comply with regulatory bodies. Video recording of trauma team resuscitations can be utilized as an effective quality improvement tool to evaluate trauma team performance, psychomotor skills and techniques, and to identify educational needs related to specific trauma populations. Video recording of Trauma resuscitations is an effective tool for improving trauma team performance by educating clinical staff regarding roles and responsibilities. PMID- 16265921 TI - Protoberberine alkaloids. PMID- 16265922 TI - The stemona alkaloids. PMID- 16265923 TI - Alkaloids of the Hernandiaceae: occurrence and a compilation of their biological activities. PMID- 16265924 TI - What have we done for you lately? Looking ahead. PMID- 16265925 TI - Having no certain answers to the much-reported crisis in health care. PMID- 16265926 TI - The ADHD challenge: how the disorder affects patients, health care providers and educators. Part one of two. PMID- 16265927 TI - Documentation done? Not done? PMID- 16265928 TI - The home health challenge: reducing acute care hospitalization. National effort aims to improve care, educate patients. PMID- 16265929 TI - Atrial fibrillation: rate versus rhythm control. PMID- 16265930 TI - Meet our members: Graham F. Greene, MD. PMID- 16265931 TI - Intensity-modulated radiation therapy (IMRT) enabling technologies. PMID- 16265932 TI - Determining the economic value of disease management programs for employers. PMID- 16265933 TI - A cost-effectiveness analysis of antibiotic therapy in macrolide-resistant community-acquired pneumonia. AB - Successful treatment of community-acquired pneumonia (CAP) can have substantial implications. As rates of antibiotic resistance of Streptococcus pneumoniae--the most common pathogen of CAP-increase, so does the likelihood that first-line pharmacotherapy will fail. Thus, the cost effectiveness and budgetary effects of treating CAP with amoxicillin/ clavulanate (AMX/CLA) extended-release (ER) and clarithromycin ER were analyzed. The model considers incidence of macrolide and AMX/ CLA-susceptible and nonsusceptible S. pneumoniae in empiric therapy. Clinical cure rates from multicenter clinical trials and published literature were used to calculate average treatment costs and success. Amoxicillin/ clavulanate ER resulted in a higher percentage of patients cured compared with clarithromycin ER (88.7% vs. 82.4%, respectively) and lower average per-patient treatment costs (dollar 437.70 vs. dollar 548.14, respectively). PMID- 16265934 TI - Addressing drug safety issues in the elderly. PMID- 16265935 TI - Preventing adverse drug reactions in the general population. AB - In 2000, the number of patient deaths attributable to adverse drug reactions (ADRs) was estimated to be 218,000 annually. More than 51% of approved drugs in the market today may have serious side effects not detected before marketing approval. The causes of ADRs are many, ranging from drug-drug interactions to simple patient noncompliance. Until the use of electronic medical records becomes ubiquitous, other partnerships must be undertaken to lower the incidence of ADRs. Health plans and pharmacy benefit managers must work together to take effective steps to increase ADR monitoring and reporting and to proactively avoid ADRs through pharmacy management tools. PMID- 16265936 TI - Changes in the newborn delivery practice and neonatal outcomes as financing changed in Los Angeles County and Orange County, California. AB - The objective of this investigation was to study changes in newborn delivery during a period of extensive changes in financing mechanisms in Los Angeles County (LAC) and Orange County (OC), California. California hospital discharge data (1990-1999) were used for the analyses. The Herfindahl-Hirschman Index (HHI) was calculated to measure the distribution of newborn deliveries among hospitals. Birth outcomes, including cesarean section rates, in-hospital deaths, interhospital transfers, and selected neonatal morbidities, were assessed. A total of 2,351,209 newborn deliveries in 124 hospitals were recorded. The number of newborn babies delivered per year decreased by 21% during the study period. A dramatic decline in the number of deliveries (up to 80%) was seen in all four LAC county hospitals. The cesarean section rate increased for Medi-Cal patients during the study period, compared with no change in the rate for privately insured patients. The newborn interhospital transfer rate for patients in LAC remained unchanged. The newborn in-hospital mortality rates for both LAC and OC decreased during the study period. PMID- 16265937 TI - Implications of the rofecoxib trials. PMID- 16265938 TI - Trends in the racial quality-of-care gap. PMID- 16265939 TI - The art of managing dementia in the elderly. AB - Dementia presents unique challenges for physicians, patients, and families, but it also offers a singular opportunity to practice the essence of the art of medicine. Elderly patients' complaints about cognition require evaluation and should never be written off as a "normal" part of aging. Dementia should be distinguished from conditions such as delirium and depression, and the type of dementia should be identified, since this will determine treatment. Treatments seek to alter the fundamental course of the disorder, to ameliorate symptoms, or to manage concomitant psychiatric and behavioral problems. Even when treatments prove ineffective, providing information and support is of great value to patients and their families and caregivers. PMID- 16265940 TI - Preventing ischemic stroke in the older adult. AB - Stroke is a deadly and disabling disease that preferentially afflicts older adults. It shares common risk factors with myocardial infarction (MI), such as hypertension, diabetes, and hyperlipidemia. Blood pressure control, cholesterol reduction with statins, and glucose control reduce the risk for both stroke and MI. Additionally, management of atrial fibrillation with warfarin reduces stroke risk. Beyond risk factor reduction, antiplatelet therapy is an effective option for lowering the likelihood of stroke in at-risk patients. Among antiplatelet agents, aspirin has been shown effective for secondary stroke prevention as well as primary and secondary MI prevention; clopidogrel for secondary stroke and MI prevention; and both ticlodipine and dipyridamole for secondary stroke prevention. Combining antiplatelet agents is rational. Carotid endarterectomy should be considered for stroke prevention in patients with ischemic symptoms; for patients with asymptomatic stenosis, potential benefit must be balanced against surgical risk. PMID- 16265941 TI - Seizures in the elderly: Nuances in presentation and treatment. AB - Acute symptomatic seizures and epilepsy are two of the most common neurologic complaints in the elderly. Stroke is the leading underlying etiology for both. Because clinical seizure manifestations in the elderly often differ from those in younger adults, they may be difficult to recognize or may be misdiagnosed. Interpretation of diagnostic tests in elderly patients with seizures is often complicated by comorbidities, and treatment decisions require careful consideration in the context of age-related physiologic changes, comorbidities, and the use of concomitant medications. Treatment of an acute seizure with a clear precipitating cause involves correcting the underlying etiology; antiepileptic drug (AED) therapy is generally reserved for patients with epilepsy (recurrent unprovoked seizures). The prognosis for elderly epilepsy patients treated with AEDs is generally good. Both older and newer AEDs are efficacious but have respective advantages and disadvantages; no ideal AED yet exists. Status epilepticus is a neurologic emergency that is particularly frequent in the elderly and associated with high mortality, although treatment can be effective. PMID- 16265942 TI - Movement disorders in the older patient: differential diagnosis and general management. AB - Movement disorders are especially prevalent in the elderly, and some are highly treatable. Because reduced agility and slowing of gait are associated with numerous movement disorders as well as with the normal aging process, the differential diagnosis of movement disorders in the elderly can be challenging. Many of these disorders share features of parkinsonism-hypokinesia, tremor, and muscular rigidity. This article reviews common and less common movement disorders in the elderly from a primary care perspective, with an emphasis on the presenting features and the differential diagnosis. It also provides general management recommendations with advice for tailoring treatment to elderly patients. PMID- 16265943 TI - Depression in older patients with neurologic illness: causes, recognition, management. AB - Depression is common in the elderly, particularly in older persons with neurologic illness. Its etiology in this population is incompletely understood and likely to be multifactorial. Identifying depression in elderly patients with neurologic illness can be a challenge, as many of its features resemble symptoms of the underlying neurologic disease or of the aging process itself. Nevertheless, recognition and effective management of depression in this population is vital, since depression is a major source of excess morbidity and since treatment often results in improved quality of life for patients and their caregivers. Assessing for suidicality is a key diagnostic consideration in this population. Antidepressant medications, psychotherapy, and electroconvulsive therapy all can be effective in treating depression in elderly neurologic patients. PMID- 16265944 TI - The primary use of venous grafts in thumb replantation. AB - The authors describe modified thumb replantation technique. With this technique it is possible to avoid extensive mobilization of the digital arteries as well as incisions, particularly at the distal thumb phalanx, and therefore reduce further damage of the amputated part. Prior to the osteosynthesis a venous graft is harvested from the distal forearm or from the thenar area. Anastomosis between the venous graft and radial digital artery on the amputated part of the thumb is completed. Osteosynthesis of the bones with Kirchner wires follows. The venous graft is further anastomosed to the dorsal venous system of the stump. The arterial system is reconstructed with an anastomosis of the ulnar digital artery with a superficially positioned volar vein of the amputated part. The replantation technique described has proved effective in all cases when it was used. PMID- 16265945 TI - Degloving injury--the use of a combination of free fasciocutaneous sensitive flap and pedicle flaps for reconstruction. AB - The authors present a case report of reconstruction after a degloving injury to hand by a combination of microsurgery and standard pedicle flaps. Degloving injury high on the hand was solved by free sensitive fasciocutaneous flap transfer from the radial forearm (Chinese flap), which covered the defect in the palm, while finger defects were covered by pedicle flaps from the abdomen. Dorsum of the hand was treated with a mesh skin graft. The authors also describe techniques of treatment and the relevant results prior to the time of microsurgery and compare them to the possibilities offered by microsurgery today, and they include comprehensive illustrations. PMID- 16265946 TI - Bronchial myoplasty with the use of latissimus dorsi muscle--a case study. AB - Dehiscence of the bronchial stump as well as the tracheo-bronchial-pleural fistula is a rare but life-threatening complication of pulmonary resections. Absence of the immediately accessible well vascularized soft tissues makes successful direct resuture of the bronchial stump or local closure of the defect problematic, particularly in the infected terrain. The authors describe a case study of successful closure of the tracheo-bronchial defect with the use of pediculated latissimus dorsi muscle in a cachectic 63-year-old patient after a right-side pneumonectomy. PMID- 16265947 TI - An unusual solution after bilateral lower extremity amputation. AB - The authors present a case of a 40-year-old male with devastating amputation trauma of both lower extremities. Reconstruction of the right lower extremity was solved by the unusual use of inserted vascularized bone-skin graft from the left crus with a simultaneous replantation of the right foot. The result after 3.5 years is preservation of one extremity with full knee mobility. The other lower extremity was fitted for thigh prosthesis. PMID- 16265950 TI - Award of the G. Whitaker International Burns Prize for 2005, Palermo, Italy. PMID- 16265948 TI - Unusual case of resistant actinomycosis following facial trauma. AB - A case of actinomycosis is reported which appeared as a complication of a ski pole injury of the face. A 16-year-old girl was treated for 10 years for resistant actinomycosis. She took long-term antimicrobial therapies, underwent repeated excisions, hyperbaric oxygen therapy, repeated skin grafting, immunological treatment, and free flap transfer. The conservative antibiotic treatment was complicated by mycosis and pseudomembranous colitis. After the defect healed up, satisfactory contour of the face was restored with additional free flap transfer. The patient has also been satisfied with the final aesthetic result. The authors highlight the importance of early diagnosis and proper antimicrobial treatment in combination with surgical excisions of granulation tissue preserving vital structures. PMID- 16265949 TI - Three-dimensional visualisation and analysis of post-operative changes in the size and shape of the dental arch and palate. AB - In craniofacial surgery and orthodontics, three-dimensional computer models of the dental arch and palate have recently entered usage in diagnosis assessment, treatment planning, case presentations and evaluation of treatment progress and outcome. In this contribution, we show how effective visualisation and evaluation of changes in the size and shape of the dental arch and palate in a given patient can be performed using superimposition of two or more 3D computer models that record the condition before and after treatment. We also present a method of three-dimensional measurement of the dental arch and palate suitable for evaluation of treatment results within retrospective and prospective studies in larger samples of subjects. PMID- 16265951 TI - Nursing in B.C. now under the Health Professions Act. PMID- 16265952 TI - A framework for professional practice. PMID- 16265953 TI - Put the care back in nursing. PMID- 16265954 TI - Feeling valued. PMID- 16265955 TI - What nurse shortage? PMID- 16265956 TI - The business of nursing. PMID- 16265958 TI - Transmitting information by E-mail and fax. PMID- 16265959 TI - Student reps turn ideas into action. PMID- 16265960 TI - Regulation and collaboration. PMID- 16265957 TI - RNs in nurse practitioner positions in rural and remote Canada. PMID- 16265961 TI - [How I treat ... lentigo maligna by topical imiquimod]. AB - Lentigo maligna is a special form of in situ cutaneous melanoma that develops on the face of sun worshipers. The topic immunostimulator imiquimod can destroy the neoplasm before it expresses its invasive potential. A strict clinical follow-up of the treated zone must be observed at least 5 years to detect any early sign of recurrence. PMID- 16265962 TI - [Ambulatory treatment of hemorrhoids]. AB - DGHAL (Doppler Guided Hemorrhoid Arterial Ligation) represents a new approach to the treatment of internal hemorrhoids; it entails exact and selective ligation of the arteries supplying the piles (hemorrhoids). The intervention can be performed on ambulatory patients under local anaesthesia. An anoscope is used which incorporates a Doppler head. The superior hemorrhoidal arteries are identified under guidance of the arterial Doppler sound and ligated through a window located just above the Doppler head. The intervention lasts some 30 minutes. Local discomfort can ensue for a few days following surgery. At one month, the time required for the internal haemorrhoids to fade away, the patient is seen again; an external hemorrhoid or residual skintag can then be considered for treatment under local anaesthesia if needed. As of November 2001 until today, more than 350 patients have been treated, and we report here on 150 of them. Long term data (6 months to 2 years) have been collected which includes 85 to 90% patient satisfaction. PMID- 16265963 TI - [Laparoscopic liver resection of a hydatid cyst]. AB - Most of the echinococcosis cases treated in Belgium are contracted in African and Mediterranean countries. In this paper the authors describe the case of a Mediterranean patient suffering from a hepatic hydatid cyst treated by oral albendazole and laparoscopic liver resection. PMID- 16265964 TI - [A bilateral dysgerminoma: a rare presentation of the Swyer syndrome]. AB - Swyer syndrome is a pure gonad dysgenesis associating 46 XY karyotype, primary amenorrhea, presence of female internal genital tract and bilateral streak gonads in a phenotypic female. The diagnosis is usually made at adolescence when the primary amenorrhea is investigated. We report the case of a 25-year-old XY woman who developed a bilateral dysgerminoma from undifferentiated gonads. PMID- 16265965 TI - [Bone quality: from theory to reality]. AB - Bone quality is an essential element of bone strength. Bone quality refers not only to bone architecture but also to its material properties which are directly dependent on bone remodeling (i.e., turnover). Therefore, maintaining bone architecture but also bone turnover is critical for optimal bone quality and for providing the ability of bone to resist fractures. PMID- 16265966 TI - [Pharmacological treatment of hypercholesterolaemia in primary prevention. The use of the different sources of information]. AB - In the first two articles of this series devoted to Evidence-Based Medicine, we have shown how to translate a clinical problem in a well formulated question and how to derive search terms from the PICO. In this article, we describe in more detail the different sources of information and how to use them. The strategy will be illustrated by answering the question whether it should be recommended to treat hypercholesterolaemia in a young woman without cardiovascular risk factors. In a following article, we will show how to adapt a search strategy to a specific question or PICO. PMID- 16265968 TI - [Analysis of pooled data from the randomized controlled trials of endarterectomy for symptomatic carotid stenosis]. AB - It is important to establish clear definitions concerning carotid artery disease, a topic that recently regained widespread interest in medical literature. Therefore, we summarize a paper of Rothwell and Barnett, recently published in the Lancet 2003 (1). In their analysis, the data of three randomized trials, comparing carotid endarterectomy to optimal medical therapy alone for a symptomatic internal carotid artery stenosis, were pooled after standardization of the measurement of the degree of stenosis and redefinition of the outcome events, to achieve comparability of the results of each trial. This analysis concerns 6092 patients with 35,000 patient-years of follow-up. Surgery offers a marginal benefit for patients with a moderate (50-69%) symptomatic carotid artery stenosis (absolute risk reduction of 4.6% for subsequent ipsilateral stroke at 5 years). Surgery is highly beneficial for patients with a symptomatic 70-99% stenosis (absolute risk reduction of 16% for ipsilateral stroke at 5 years). The benefit is uncertain for a symptomatic "near occlusion" (99% with retarded opacification of the distal internal carotid artery): absolute risk reduction of 1.7% for ipsilateral stroke at 5 years). PMID- 16265967 TI - [Analysis of two year heroin seizures in the Liege area]. AB - The results of heroin analysis from seizures in the Liege area during the last two years are presented in this article. Between January 2003 and January 2005, 50 samples were analysed in the Laboratory of Clinical Toxicology and Forensic Toxicology of the University of Liege. Mean heroin concentration was 14,7%. Noscapine and papaverine, other opium alcaloids, were simultaneously present with heroin. As diluents, we only identified caffein and acetaminophen. PMID- 16265969 TI - [How I prevent ... minor acne]. AB - Acne is a disorder affecting the majority of subjects during adolescence that may persist at various grades of severity during adulthood. This disorder may manifest itself at different levels of severity ranging from discrete to severe. The lesions may be sporadic or follow a wild course. The therapeutic strategy depends on these progression characteristics and must fit to each individual patient. This review focuses on the minor but the most frequent types of acne that may benefit from dermocosmetic treatments. PMID- 16265970 TI - [Food intolerance and allergy in the irritable bowel syndrome]. AB - Irritable Bowel Syndrome (IBS) is a complex and heterogeneous entity that concerns about 1/4 of adults and would be responsible for 50% of gastro enterology medical consultations. IBS etiopathogenesis and physiopathology are not yet fully known. Implications of food intolerance and allergy in this syndrome remain controversial. In this review we recall numerous mechanisms of allergy or food intolerance generating IBS, as well as clinical entities mimicking an IBS. We present a series of diagnostic tests and potential treatments. We finally propose a management strategy for these patients. PMID- 16265971 TI - Employee assistance programs in the new millennium. AB - This article presents an overall view of Employee Assistance Programs (EAPs). Beginning with the history, this article describes various models and essential ingredients of EAPs. It then discusses current trends including integration with Work/Life, web-based services, EAP accreditation, and the growth of international programs. Several issues are discussed including the need for licensing done nationally rather than by states and the effect the lack of substance abuse and brief counseling education has had on the delivery of EAP services. Future directions for EAPs emphasize the importance of quality assurance and the development of performance measurements, performance guarantees, and outcome measurements. The formation of the Alliance for Employee Assistance Advancement, an organization of organizations, completes the description and shows a new direction for EAPs in the next millennium. PMID- 16265972 TI - Workplace disaster preparedness and response: the employee assistance program continuum of services. AB - Response programs for workplace critical and traumatic events are becoming an acknowledged and sought after standard of care. The current trauma literature recognizes what goes on in the workplace between the Employee Assistance Program (EAP) and management. The authors have taken this intra-organizational relationship, assimilated the information, and developed a model that recognizes and supports management throughout the continuum of response to workplace traumatic events. The model recognizes the EAP as an important workplace resource and tool in management's ability to strike the balance of managing the workforce while assisting in recovery following workplace trauma. The introduced concept defines the continuum and highlights the before, during, and after phases, showing how EAP supports management in most effectively doing their job. PMID- 16265973 TI - A federal perspective on EAPs and emergency preparedness. AB - Federal Employee Assistance Programs (EAPs) have a long history of intervention in emergency situations, and their role has expanded since September 11, 2001. There is considerable evidence on the importance of organizational factors such as social support in protecting disaster victims from the psychological effects of trauma. The authors recommend that EAPs become integrally involved in organizations' emergency planning processes, so that such supports can be built into all aspects of plans. EA professionals should function as organizational consultants, not simply as helpers who will eventually care for those affected. Two federally based case studies provide examples of EA professionals who have successfully used such an approach in their respective organizations. PMID- 16265974 TI - Fostering human continuity: an essential element in workplace crisis intervention. AB - News from throughout the world illustrates how disasters are impacting people during their commute and at the workplace. Now more than ever employers must ensure that personnel have the tools to deal with the impacts of these critical events. This article outlines how organizations can build psychological infrastructure in their corporate culture to foster resilience in the workplace. PMID- 16265975 TI - Organizational crisis management: the human factor. AB - While many professionals are quite competent when dealing with operational aspects of organizational continuity, often the "human factor" does not receive adequate attention. This article provides a brief overview of a soon to be published book by the same title. It provides a comprehensive understanding of the ubiquitous yet complex reactions of the workforce to a wide array of organizational disruptions. It goes beyond the short term intervention of debriefings to describe the more extensive pre and post incident strategies required to mitigate the impact of crises on the workforce. It is important to remember: "An organization can get its phone lines back up and have its computers backed up...but its workers may still be messed up." PMID- 16265976 TI - Bioterrorism and its aftermath: dealing individually and organizationally with the emotional reactions to an anthrax attack. AB - From September 2001 through April 2004, the United States Postal Service (USPS) dealt, for the first time, with bioterrorism resulting in employee deaths and the closure of a large mail processing plant in Washington, D.C. The Employee Assistance Program (EAP) partnered with the USPS throughout this tumultuous time to meet the multiple and evolving behavioral health needs of the employees and facilitate the employees' emotional preparedness for their return to work at the closed facility. This paper discusses the reactions manifested by the employees during this extended period, as well as the EAP activities in the recovery process. PMID- 16265977 TI - Managing trauma in the South African mining industry. AB - Deep sub-surface mining is one of South Africa s leading industries. It is also one of the most dangerous. While safety is maintained as much as possible to minimize cave-ins and other accidents, underground tremors and earthquakes are commonplace and unpredictable and usually involve life threatening injuries and loss of life. Crisis intervention and trauma management are essential to helping workers cope with mining accidents. This article describes the trauma management program developed by the employee assistance program at the Chamber of Mines in South Africa to respond to workplace accidents. PMID- 16265978 TI - The development and evaluation of an internal workplace violence risk assessment protocol: one organization's experience. AB - The creation and development of a Risk Assessment Team at a large urban university is presented as a case study, with particular focus on the role the Employee Assistance Program (EAP) plays in the multidisciplinary team. The structure of the team and differing roles and responsibilities of members will be discussed. A specific protocol for addressing incidents will be introduced, along with changes in the team ' response over time. Major lessons learned will be presented, as well as the challenges the team faces today, and discussion of areas for future research and evaluation. PMID- 16265979 TI - Impact of the September 11 attack on flight attendants: a study of an essential first responder group. AB - This paper discusses a study (funded by the Center for Substance Abuse Prevention) of the health/mental health and work-related well-being of flight attendants in the aftermath of September 11. Flight attendants, as an occupational group, had a distinctive exposure to September 11. In addition to work-related exposure in the immediate aftermath of the attacks, flight attendants have experienced major and ongoing changes in their work environment and job description and many have been exposed to potentially traumatic incidents on the job. Analysis of survey and focus group data from flight attendants in the Association of Flight Attendants showed high reported stress and related mental health and behavioral impacts among flight attendants since September 11. A significant new finding is that the effect of continued trauma in the flight attendants' work environment impacted their ability to recover from the original trauma associated with the events of that day. This study highlights the role of the after-effects of a traumatic event on trauma response and suggests that direct exposure, as traditionally defined, is not necessarily a primary mediating factor in trauma response for this occupational group. PMID- 16265980 TI - Enhancing relationships: peer teams and mental health professionals. AB - Employee Assistance Programs and Medical Benefits Plans can provide peer teams and organizations access to a large number of mental health professionals. Although the peer team and the organization may have a limited number of mental health professionals whom they prefer to utilize exclusively, there may be a need for additional mental health resources in certain instances. This article discusses developing relationships between mental health providers and peer teams. A case study is presented that describes a process that allows teams to enhance and integrate the full resources and services of mental health providers and/or Employee Assistance Programs available to them. PMID- 16265981 TI - Revival of a tradition of Chinese medicine in a reclaimed Chinese territory. AB - The effect of the political transition from a British Colony to a Special Administration Region of China in 1997 on the tradition of Chinese medicine is examined using historical reviews as well as interviews with various sectors of the population in Hong Kong. Results show that the political change has stimulated the formation of a location-specific Hong Kong Chinese medicine strongly characterized by both scientific and commercial elements developed from the culture of Chinese medicine. PMID- 16265982 TI - Chai-hu-gui-zhi-gan-jiang-tang regulates plasma interleukin-6 and soluble interleukin-6 receptor concentrations and improves depressed mood in climacteric women with insomnia. AB - This study was performed to compare the effects of Chai-hu-gui-zhi-gan-jiang-tang (Saiko-keishi-kankyo-to), an herbal medicine, in improving depressed mood and on plasma interleukin-6 (IL-6) and soluble interleukin-6 receptor (sIL-6R) concentrations with those of anti-depressants in peri- and post-menopausal women. Ninety patients complaining of menopausal symptoms including insomnia who were diagnosed with mood disorder based on DSM-IV were recruited and separated into two groups (Chai-hu-gui-zhi-gan-jiang-tang group was selected on the basis of SHO for 42 cases, while anti-depressants were used for 48 cases), and plasma IL-6 and sIL-6R concentrations were determined before and after three months of the treatment. There were no significant differences in the decrease in both climacteric and Hamilton depression score after treatment between the two groups. Plasma IL-6 and sIL-6R concentrations were significantly lower in the Chai-hu-gui zhi-gan-jiang-tang group (-34.8 +/- 15.5% and -22.4 +/- 14.6%, respectively) than in the anti-depressant group (7.5 +/- 4.8% and 2.4 +/- 3.8%, respectively) after 3 months of treatment. Correlations between rate of decrease in climacteric score and plasma IL-6 (R = 0.498, P = 0.0056) and sIL-6R (R = 0.512, P = 0.0045) concentrations were observed. Chai-hu-gui-zhi-gan-jiang-tang reduced plasma IL-6 and sIL-6R concentrations in relation to improvement of depressed mood during treatment. The findings of this study suggest that Chai-hu-gui-zhi-gan-jiang-tang has the potential to decrease morbidity by alleviation of stress reactions in peri- and post-menopausal women. PMID- 16265984 TI - Differences in electrical conduction properties between meridians and non meridians. AB - Therapy using the acupuncture meridian system is an important part of traditional Chinese medicine. The purpose of this study was to investigate the electrical conduction properties of the meridians. The current conduction and potential profiles were compared after switching the current direction in the Hegu (LI-4) and Quchi (LI- 11) meridians and over a non-acupuncture point 1 cm from Quchi (LI 11) in 20 healthy subjects. Both meridians demonstrated significantly higher conductivity between Hegu (LI-4) and Quchi (LI-11) than between Hegu (LI-4) and the non-acupuncture point. The direction of current, peak frequency and absolute potential values in the direction Hegu (LI-4) to Quchi (LI-11) differed significantly from those in the direction Quchi (LI-11) to Hegu (LI-4). These results suggest that the conducting pathways are stronger in the meridians than in the non-meridians and that preferential conduction directions exist between two acupuncture points. These results are consistent with the theories of Qi circulation and traditional Chinese medicine. PMID- 16265983 TI - Clinical trial of herbal formula on weight loss in obese Korean children. AB - A traditional Korean herbal formula (KH), which is based on Taeumjowi-tang, is currently the most widely used herbal formula in Korea. In this study, KH was administered to obese children for 30 days, and was found to be clinically safe and effective. The subjects were children admitted to hospital to be treated for obesity with relative body weights (%RBW) of 20% or more. Originally, there were 31 subjects, but nine dropped out during the experiment. There were eight girls and 14 boys, whose average age was 11.00 +/- 2.62 years, average weight was 53.37 +/- 17.29 kg, and average period (30-day amount) of KH dosage was 51.18 +/- 22.58 days. The short-term effects of KH on obese children were the reduction of their BMI from 24.34 +/- 3.10 to 23.26 +/- 3.00 kg/m2, of %RBW from 34.41 +/- 10.90 to 25.94 +/- 11.18% (p < 0.01), of body fat mass from 17.99 +/- 5.37 to 16.50 +/- 4.82 kg, and of body fat from 34.16 +/- 3.75 to 32.08 +/- 3.15% (p < 0.01). Concerning anthropometrical measurements, abdominal skin-fold decreased from 26.16 +/- 9.08 to 22.90 +/- 8.35 mm, as did subscapular skin-fold from 20.86 +/- 5.20 to 18.46 +/- 5.31 mm (p < 0.01). In terms of serum lipid levels, which are indices of heart disease, their total cholesterol decreased from 195.38 +/- 31.39 to 183.25 +/- 33.27 mg/dl, the arteriosclerosis index from 4.100 +/- 0.81 to 3.84 +/- 0.64 mg/dl (p < 0.05), and serum leptin level from 14.91 +/- 6.59 to 12.24 +/ 4.98 ng/ml (p < 0.01). Concerning the safety of KH, there were no significant changes in the subjects' livers, hearts, or kidneys. Nor were there any short term signs of clinically serious side effects or withdrawal symptoms observed. The short-term effects of KH on obese children are weight loss and a decrease in obesity. PMID- 16265985 TI - Evaluation of the cun measurement system of acupuncture point location. AB - Locating acupuncture points reliably and reproducibly is indispensable for the scientific research of acupuncture and for assuring the best care of patients. Unreliable point location can produce confounding results for acupuncture research and clinical practice. Two traditional methods of point location are currently used, directional (F-cun) and proportional (B-cun) methods, which are collectively called the cun measurement system. Reports have been published on the validity of the cun measurement system in Australian subjects; however, as acupuncture originated in ancient East China, it is possible that anthropometric data may differ in Asian people and other races. Therefore, we measured anthropometric data according to the cun measurement system in contemporary Korean patients. The F-cun measurements were significantly different from the B cun measurements and varied significantly according to the arbitrarily selected F cun standard. In addition, we observed further differences of F-cun measurements in the extremities of obese subjects. We concluded that the F-cun method is unreliable and that further research should be conducted to determine a more accurate point-locating method primarily based on the B-cun method. PMID- 16265986 TI - Effects of chai hu (radix burpleuri) containing formulation on plasma beta endorphin, epinephrine and dopamine on patients. AB - Chai Hu (Radix Burpleuri), a major ingredient in many traditional Chinese medicine formulas, such as Xiao Yan Wan, is used in the treatment of liver stagnation and spleen deficiency syndrome (LSSDS). The objectives of this study were to examine the effects of Xiao Yao Wan containing Chai Hu on the changes of plasma indices in patients with LSSDS. Fifty-eight cases of LSSDS were randomly divided into two groups: 41 cases in the experimental group were treated with Xiao Yao Wan containing Chai Hu and 17 cases in the control group were treated with Zhi Bai Di Huang Wan for one consecutive month in a single blind design. Before and after treatment, high performance liquid chromatography (HPLC) was applied to determine the changes of plasma norepinephrine (NE), epinephrine (E) and dopamine (DA). Radioimmunoassay was performed to measure the amount of plasma beta-endorphin (beta-EP), adrenocorticotropin hormone (ACTH), estradiol (E2) and testosterone (T), and laser nephelometry was also conducted to measure plasma immunoglobulin A (Ig A) and G (Ig G). Compared to baseline levels, plasma beta-EP was significantly increased (p < 0.01), while E and DA were markedly decreased (p < 0.01) after the administration of Xiao Yao Wan in the experimental group. The other indices did not change. This is the first evidence showing that the effect of Xiao Yao Wan containing Chai Hu on the treatment of patients with LSSDS may be through enhancing plasma beta-EP and decreasing E and DA release. We conclude that Xiao Yao Wan containing Chai Hu regulates nervous and endocrine systems and contributes to the improvement of the clinical status of patients with LSSDS. PMID- 16265987 TI - Antioxidant and antiplatelet effects of dang-gui-shao-yao-san on human blood cells. AB - Dang-Gui-Shao-Yao-San (DGSYS) is a mixture of medicinal herbs, which has long been used in traditional Chinese medicine for treating anemia and ovulary disorders. Its preparation comprises Angelicae sinensis (Oliv.) Diels, Ligustucum chuanxiong Hort, Paeonia lactiflora pall, Poria cocos (Schw.) Wolf, Atractylodis macrocephala Koidz and Alisma orientalis (Sam.) Juzep. The present study examined the anti-superoxide formation, free radical scavenging and anti-lipid peroxidation activities of DGSYS by xanthine oxidase inhibition, cytochrome C system with superoxide anion released by the fMLP or PMA activating pathway in human neutrophils, and FeCl2 ascorbic acid-induced lipid peroxidation effects on lipids in rat liver homogenate, respectively. DGSYS showed anti-superoxide formation and free radical scavenging activity in a concentration-dependent manner. It also inhibited PMA- but not fMLP-induced superoxide anion released from human neutrophils. These antioxidant actions of DGSYS showed beneficial cytoprotective effects against lipid peroxidation in rat liver homogenate, human platelet aggregation induced by arachidonic acid (AA) and adenosine diphosphate (ADP) and mitomycin C-mediated hemolytic in human erythrocytes. PMID- 16265988 TI - Birdcage model for the Chinese meridian system: part VI. meridians as the primary regulatory system. AB - It is imperative to define the fundamental concepts of Qi, channels, and the meridian system of Chinese medicine in terms of scientific terminology before any meaningful and mutually beneficial dialog can begin between Chinese and Western medicine. In the Chinese theory, the meridian system as a whole is the system of the body. We propose the existence of a meridian regulatory system that governs interactions between and adjusts functions of internal organs, connects them to the body surface through a network of pathways (channels) and displays their status on the skin. The meridian systems is analyzed as a 28-leg, uniform, low pass birdcage coil, where each leg represents a channel. The channel is analyzed as a transmission line and Qi is the standing wave riding on it. Each segment in the channel is represented as a section of the transmission line and it is in natural oscillation, with its second lowest resonant frequency being the 50-round circulation frequency f50, 0.578 x 10(-3) Hz. PMID- 16265989 TI - Dynamic change in energy metabolism by electroacupuncture stimulation in rats. AB - The electrical stimulation of meridian points in rats inhibits the withdrawal reflex of the nociceptive tail. Its pain mechanisms are well-documented. Moreover, electroacupuncture (EA) at special abdominal acupoints has been shown to induce a short-term hypoglycemia effect in streptozotocin diabetic rats. The Zusanli and Zhongwan acupoints have been widely used in traditional Chinese medicine to relieve symptoms of diabetes mellitus. It is still unclear whether they can affect extracellular glucose and lactate metabolites at the cellular level. The aim of this study is to evaluate these effects using a rat model for the analysis of extracellular neurochemicals. First, electrical stimulus of 2 ms 2 Hz square pulses (30 minutes) was applied to anesthetized intact rats (n = 7) at the Zusanli points. One and a half hours later, a second electrical stimulus (2 Hz pulses, 30 minutes) was delivered to two of the rats at the same spot. Another two rats received a different stimulation (100 Hz pulses, 30 minutes) at the same location. In the final three rats, a second electrical stimulus of 2 Hz pulses was delivered to non-acupoints. An automated micro-blood sample collector was used to examine the glucose, pyruvate and lactate concentrations. The EA signal has an influence on the biologic process of energy metabolism by mediating dynamic extracellular neurochemical changes. The EA at limb acupoints of the lower limbs induces a decrease in glucose, an increase in lactate metabolites and a decrease in the lactate/glucose ratio. Moreover, the increased lactate/glucose ratio suggests that the cell has an increased anaerobic glucose metabolism. PMID- 16265990 TI - Protective effect of propolis ethanol extract on ethanol-induced renal toxicity: an in-vivo study. AB - Acute p.o. administration of absolute ethanol (10 ml/kg) to fasted mice would produce extensive renal failure. Pretreatment with p.o. administration of propolis ethanol extract (PEE) could prevent such renal failure effectively and dose dependently. This renal protective effect of PEE may be contributed, at least in part, to its antioxidative activity. The maximal antioxidative effect against absolute ethanol (AE)-induced renal failure could be observed 1 hour after PEE administration. In order to further investigate the renal protective mechanism of PEE, lipid peroxidation and superoxide scavenging activity were conducted in vivo. PEE exhibited dose-dependent antioxidative effects on lipid peroxidation in mice renal homogenate. Results indicated that mice with acute renal failure have higher malonic dialdehyde (MDA) levels compared with those in PEE administered mice. It was concluded that the renal protective mechanism of PEE could be contributed, at least in part, to its prominent superoxide scavenging effect; hence, it could protect, indirectly, the kidney from superoxide-induced renal damages. PMID- 16265991 TI - Traditional Chinese herbs against hypertension enhance memory acquisition. AB - Recent findings of a link between high blood pressure (BP) and dementia have given new prospects. The aim of this study is to analyze a mixture of Chinese herbs, Tianma Gouteng Decoction (TGD), which was traditionally used to treat hypertension, and investigate its relation to ameliorating cognitive impairment. We discovered that TGD also had properties involving enhancement of memory acquisition (learning) skills in mice, but not memory consolidation. It was observed that TGD could prolong the step-through latency at doses of 1.0 and 2.5 g/kg on passive avoidance task in mice. TGD could be developed further to treat mice with amnesia, which was induced by scopolamine at the same dose under long term (8 days) administration. PMID- 16265992 TI - Effects of saponin monomer 13 of dwarf lilyturf tuber on L-type calcium currents in adult rat ventricular myocytes. AB - The saponin monomer 13 of dwarf lilyturf tuber (DT-13), one of the saponin monomers of dwarf lilyturf tuber, has been found to have potent cardioprotective effects. In order to investigate the effect of DT-13 on L-type calcium currents (I(Ca,L)), exploring the mechanisms of DT-13's cardioprotective effects, we directly measured the I(Ca,L) in the adult rat cardiac myocytes exposed to DT-13 using standard whole-cell patch-clamp recording technique. Our results showed that DT-13 exerted inhibitory effects on the I(Ca,L) of the single adult rat cardiac myocytes. The current density was reduced by about 38% after exposure of the cells to DT-13 (0.1 microM) for 10 minutes, from the control value of 7.46 +/ 1.31 pA/pF to 4.25 +/- 0.35 pA/pF (n = 6, p < 0.05). This I(Ca,L)-inhibiting action of DT-13 was concentration-dependent. DT-13 up-shifted the current-voltage (I-V) curve, but did not significantly affect the half activation potential (V0.5). V0.5 was from -11.8 +/- 0.9 mV in the control to -12.6 +/- 1.9 mV in the presence of DT-13 at 0.1 micromol/L. DT-13 at 0.1 microM did not markedly affect the activation of I(Ca,L), but shifted the inactivation curve of I(Ca,L) to the left. In combination with previous reports, these results suggest that there might be a close relationship between the cardioprotective effects of dwarf lilyturf tuber and the inhibitory effects of DT-13 on L-type calcium currents. PMID- 16265993 TI - Effects of ganoderma lucidum extract on chemotherapy-induced nausea and vomiting in a rat model. AB - Chemotherapy is highly cytotoxic, causing a number of severe adverse effects such as nausea and vomiting. Herbal medicines, which can often be used on a daily basis for prolonged treatment, may be clinically beneficial. Ganoderma lucidum or Lingzhi mushroom has been recognized as a remedy in treating a number of medical conditions, including balancing immunity and decreasing drug-induced side effects. It has been shown that rats react to emetic stimuli, like the chemotherapy agent cisplatin, by increased consumption of kaolin, known as pica; and this rat model has been utilized to evaluate novel anti-emetic compounds. In this study, we evaluated the effects of a G. lucidum extract (SunRecome, the most commonly used Lingzhi mushroom extract in China) in attenuating cisplatin-induced nausea and vomiting in the rat pica model. We observed that intraperitoneal cisplatin injection caused a significant increase in kaolin intake at 24, 48, 72 and 96 hours, reflecting cisplatin's nausea and vomiting action. This cisplatin induced kaolin intake dose-dependently decreased after 1, 3 and 10 mg/kg G. lucidum extract injection (p < 0.01). In addition, there was a significant reduction of food intake after cisplatin. The cisplatin-induced food intake reduction improved significantly after G. lucidum extract administrations in a dose-related manner (p < 0.01), suggesting a supportive effect of the extract on general body condition. Future controlled clinical trials are needed to evaluate the safety and effectiveness of this herbal medication. PMID- 16265994 TI - Activity of wen-pi-tang, and purified constituents of rhei rhizoma and glycyrrhizae radix against glucose-mediated protein damage. AB - Wen-Pi-Tang, an Oriental medical prescription composed of Rhei Rhizoma, Ginseng Radix, Aconiti Tuber, Zingiberis Rhizoma and Glycyrrhizae Radix, is used clinically as a medicine to treat renal failure. This study was conducted to examine the inhibitory activity of the five crude drug components of Wen-Pi-Tang and several pure compounds isolated from Rhei Rhizoma and Glycyrrhizae Radix against the protein glycation reaction. Rhei Rhizoma exerted the most potent activity, Zingiberis Rhizoma and Glycyrrhizae Radix showed relatively moderate activity, whereas Aconiti Tuber and Ginseng Radix showed weak activity. On the other hand, of 20 compounds obtained from Rhei Rhizoma and Glycyrrhizae Radix, tannins, especially rhatannin, RG-tannin and procyanidin B-2 3,3'-di-O-gallate, showed significantly strong activities that were more effective than the positive control, aminoguanidine. Some flavones such as licochalcone A and licochalcone B, and anthraquinones such as emodin and aloe-emodin, also showed inhibitory activity. These findings may help to explain, at least in part, certain pharmacological activities of Wen-Pi-Tang, whose clinical efficacy against renal failure is already recognized. PMID- 16265995 TI - Isolation and structure elucidation of a new potent anti-neoplastic diterpene from Dendrostellera lessertii. AB - Two diterpene esters were isolated from Dendrostellera lessertii. These compounds were identified as compound I (12-O-benzoyl-3,5-hydroxy-6,7-epoxy-resiniferonol 9,13,14-orthobenzoate) and compound 11 (12-O-benzoyl-5-hydroxy-6,7-epoxy resiniferonol-9,13,14-orthodecanoate). Cytotoxicity evaluation of these two compounds, using seven different cancerous cell lines, indicated that compound I with IC50 of 5-25 nmol, is 2.5 times more active than compound II. Using flow cytometry technique, it was found that treatment of the most responsive cells (K562) with compound I inhibited the progression of cells through G1 phase by almost 20% compared to the untreated cells. PMID- 16265996 TI - Environmental tobacco smoke: science, policy, and controversy. PMID- 16265997 TI - Outcomes in carpal tunnel syndrome: symptom severity, conservative management and progression to surgery. AB - PURPOSE: This study investigated the relationship between severity of symptoms and success of nonoperative and operative treatment in patients with carpal tunnel syndrome (CTS). METHODS: An observational cohort study regarding the management of CTS was conducted. Thirty patients referred to a tertiary hand centre with a diagnosis of CTS were prospectively followed. Twenty-five of the patients (47 affected hands) were available for long-term follow up to determine management outcomes. Self-report symptoms and physical impairments were assessed and documented at baseline, 6 weeks, and 12 weeks using the CTS Severity Score (SSS), the Disability-Shoulder, Arm and Hand Score (DASH), and the Levine Functional Score. Longer-term follow-up was conducted to identify status on outcome measures and whether patients proceeded to surgery. RESULTS: Those who proceeded to surgery (n = 27/47 hands) had higher initial CTS SSS and DASH scores and also maintained higher scores compared to those who improved with conservative management (p < 0.05). Improvements occurred in the SSS (P < 0.0001), Functional Score (P < 0.001), and DASH score (P < 0.05) following surgery in the patients resistant to conservative management. Recovery of grip and dexterity was less satisfactory. DISCUSSION: This study suggests that the SSS is useful in the triage of patients on surgical wait-lists as patients with high initial scores or failure to change in short-term follow-up are likely to proceed to surgical release. Despite prolonged symptoms and previous treatment, patients with lower SSS scores had moderate success with a second trial of conservative management. PMID- 16265999 TI - Agenesis of the corpus callosum: lessons from humans and mice. AB - BACKGROUND AND PURPOSE: The corpus callosum serves as a bridge to associate fibres between the two cerebral hemispheres. In placental mammals, this commissure provides for higher order neurological advantages. The molecular pathways involved in the development and pathogenesis of accallosal defects are sparse. The article reviews the current progress of studies undertaken to discern the embryological and genetic basis of the development of the corpus callosum. SOURCES OF DATA: The literature, including from sources such as MEDLINE and OMIM, were subjected to searches for articles reporting findings on corpus callosum development in humans and mice. PRINCIPAL FINDINGS: At least forty-six malformation syndromes and metabolic disorders have been reported in patients with complete agenesis or hypoplasia (dysgenesis) of the corpus callosum. Thirteen of these syndromes have an unknown mode of genetic inheritance, and the remaining syndromes and metabolic disorders exhibit either autosomal or X-linked inheritance among affected families. The use of patients with accallosal defects have identified mutations in at least thirty genes of the human genome, and therefore with roles implicated in the development of the corpus callosum. Patients with chromosome aberrations have been useful in defining regions on chromosomes that contain candidate genes for the development of the corpus callosum. At least eighteen different human chromosomes with numerical and/or structural aberrations have been reported in patients with acallosal defects. The mouse is an excellent model to study the structural and genetic factors that influence the development of the corpus callosum, with many similarities evident in humans. Spontaneous development of acallosal defects has been reported in at least seventeen mouse strains. Furthermore, with the use of Genetically Engineered Mice, a minimum of 15 candidate callosal agenesis genes were modeled in order to provide insightful knowledge of the molecular-structural parameters required for development of the corpus callosum. Of these mice, six had complete true agenesis of the corpus callosum, five had either true agenesis or hypoplasia of the corpus callosum, and four had hypoplasia of the corpus callosum. CONCLUSIONS: The molecular mechanisms implicated in the pathogenesis of agenesis or hypoplasia of the corpus callosum are at the verge of discovery, and are challenged by the complexity of many genes involved. Despite these barriers, findings from a complementary human-mouse model system have been helpful in understanding the genetic (molecular) causes of accallosal defects, a fascinating phenotype for over a century. PMID- 16265998 TI - Prevalence of cervical human papillomavirus in Taiwanese women. AB - PURPOSE: To define the prevalence rate of cervical human papilloma virus (HPV) using DNA oligonucleotide microarray and its correlation with risk factors in Taiwanese women in metropolitan Taipei. METHODS: Thirteen hundred and twenty healthy women, aged 21 - 65 yr without history of cervical intraepithelial neoplasia (CIN) or carcinoma were included in this prospective study. Pap smear and HPV typing using oligonucleotide microarray were performed for each woman. They were given a standardized questionnaire to obtain information about the risk factors of cervical cancer in Taiwan. RESULTS: The overall HPV positivity was 19.85% and multiple infections were found in 35.84% of the infected group, 7.92% of the whole study population. The younger the subject, the higher was the infection rate and multiple infection rates. The most common HPV types were 16, 18, 58, 52, 51 and 56, which is different from the western world. The sensitivity of the HPV DNA chip in detecting CIN and cervical carcinoma is 97.06%, and 100% in detecting CIN 2 or more lesions. Risk factors for HPV infection include earlier coitarche (P < 0.01), multiple sexual partners (P < 0.05), history of sexually transmitted disease (P < 0.05), two or more vaginal deliveries (P < 0.05) and infrequent use of condoms (P < 0.05). The association between oral contraception or cigarette smoking and HPV infection could not be determined because few women smoke or used oral contraception. There was no relationship between induced abortion and HPV infection. CONCLUSIONS: About one-fifth of adult women in metropolitan Taipei were cervical HPV positive. The popular HPV types and the risk factors of HPV infection in metropolitan Taipei are not the same as those in the western world. The sensitivity of the HPV DNA chip in detecting cervical neoplasia is very high. PMID- 16266001 TI - Laboring in the U.P. PMID- 16266002 TI - Protecting our current and future RNs. PMID- 16266000 TI - Frequently asked questions regarding hours of work and wages. PMID- 16266003 TI - 101 ways to improve nursing culture: respect diversity. PMID- 16266004 TI - Hurricane Katrina. Katrina's other victims. PMID- 16266005 TI - Veterinary colleges boost relief efforts. PMID- 16266006 TI - Clinics-only Web area a new resource for pet wellness campaign. PMID- 16266007 TI - Medicare to introduce prescription drug plan. PMID- 16266008 TI - Additional comments on pharmacokinetics of fluoxetine. PMID- 16266009 TI - Veterinary schools and the profession: a search for bearings in the new century. PMID- 16266011 TI - Employment of female and male graduates of US veterinary medical colleges, 2005. PMID- 16266010 TI - What is your diagnosis? Primary neoplasia of the stifle joint and, less likely, fungal osteomyelitis. PMID- 16266012 TI - Reliability of using reagent test strips to estimate blood urea nitrogen concentration in dogs and cats. AB - OBJECTIVE: To evaluate the clinical accuracy of reagent test strips used to estimate BUN concentration in dogs and cats. DESIGN: Prospective study. ANIMALS: 116 dogs and 58 cats. PROCEDURE: Blood samples were collected at the time of admission to the hospital. Estimates of BUN concentration obtained with reagent test strips (category 1 [5 to 15 mg/dL], 2 (15 to 26 mg/dL], 3 [30 to 40 mg/dL], or 4 [50 to 80 mg/dL]) were compared with SUN concentrations measured with an automated analyzer. For dogs, category 1 and 2 test strip results were considered a negative result (nonazotemic) and category 3 and 4 test strip results were considered a positive result (azotemic). For cats, category 1, 2, and 3 test strip results were considered a negative result (nonazotemic) and category 4 test strip results were considered a positive result (azotemic). RESULTS: On the basis of SUN concentration, 40 of the 174 (23%) animals (20 dogs and 20 cats) were classified as azotemic. One dog and 2 cats had false-negative test strip results, and 1 dog had a false-positive result. Sensitivity and specificity were 95% (20/21) and 99% (94/95), respectively, for dogs and 87% (13/15) and 100% (43/43), respectively, for cats. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that reagent test strips are a reliable method for rapidly estimating BUN concentrations in dogs and cats. Because test strip results are only semiquantitative and there remains a potential for misclassification, especially in cats, urea nitrogen concentration should ultimately be verified by means of standard chemistry techniques. PMID- 16266013 TI - Severe bronchoconstriction after bronchoalveolar lavage in a dog with eosinophilic airway disease. AB - Bronchoalveolar lavage (BAL) is considered to be a safe procedure in humans and other animals. However, in some instances, potentially life-threatening complications can arise. In this clinical report, a dog with eosinophilic airway disease that underwent a BAL and subsequently became profoundly dyspneic during the recovery period is described. The dyspnea was severe enough to warrant mechanical ventilation for almost 24 hours. Several anti-inflammatory and bronchodilatory medications were also used. The dog was successfully weaned off the ventilator and made a full recovery. On the basis of radiographic findings and clinical response to treatment, we believe the dog had acute exacerbation of eosinophilic airway disease and severe bronchoconstriction secondary to the BAL. Caution should be exercised when performing a BAL if there is suspicion of a reactive airway disease. PMID- 16266014 TI - Sensory and motor neuropathy in a Border Collie. AB - A 5-month-old female Border Collie was evaluated because of progressive hind limb ataxia. The predominant clinical findings suggested a sensory neuropathy. Sensory nerve conduction velocity was absent in the tibial, common peroneal, and radial nerves and was decreased in the ulnar nerve; motor nerve conduction velocity was decreased in the tibial, common peroneal, and ulnar nerves. Histologic examination of nerve biopsy specimens revealed considerable nerve fiber depletion; some tissue sections had myelin ovoids, foamy macrophages, and axonal degeneration in remaining fibers. Marked depletion of most myelinated fibers within the peroneal nerve (a mixed sensory and motor nerve) supported the electrodiagnostic findings indicative of sensorimotor neuropathy. Progressive deterioration in motor function occurred over the following 19 months until the dog was euthanatized. A hereditary link was not established, but a littermate was similarly affected. The hereditary characteristic of this disease requires further investigation. PMID- 16266015 TI - Placentitis associated with leishmaniasis in a dog. AB - A 1.5-year-old Coonhound from Maryland aborted 7 fetuses. Placenta and internal tissues of 1 fetus were examined histologically. The predominant lesion was placentitis characterized by necrosis and infiltration of mixed leukocytes. Numerous Leishmania spp amastigotes were identified in placental trophoblasts, and the diagnosis was confirmed by use of immunohistochemical staining with Leishmania-specific antibodies. Protozoa were not found in the fetal tissues. An indirect fluorescent antibody test yielded a serum titer of 1:100, and a recombinant K39 immunoassay of serum yielded positive results for the K39 Leishmania antigen. PMID- 16266016 TI - Foramen magnum decompression for treatment of caudal occipital malformation syndrome in dogs. AB - A method for foramen magnum decompression (FMD) in dogs with caudal occipital malformation syndrome (COMS) and results for 16 dogs are described. In brief, a dorsal approach to the caudal portion of the occiput and arch of the atlas was made, and a high-speed drill was used to remove a portion of the occiput in the region of the foramen magnum and the dorsal aspect of C1. The meninges that were exposed were removed or marsupialized to surrounding tissues. Foramen magnum decompression was performed in 16 dogs. No intraoperative complications occurred, and postoperative complications occurred in only 2 dogs after initial surgery and in 1 of these dogs after follow-up surgery. In both dogs, postoperative complications after the initial surgery resolved without additional treatment. One dog was nonambulatory tetraparetic after follow-up surgery and died of a suspected ruptured viscus 9 days after surgery. Four dogs developed evidence of scar formation at the surgery site and required additional surgery. Overall, 14 dogs survived, 1 died, and 1 was euthanatized. Clinical signs resolved in 7 of the 14 dogs that survived, improved in 6, and did not change in 1. Results suggest that FMD may be an effective treatment for dogs with COMS, especially if performed early in the course of the disease. PMID- 16266017 TI - Neonatal isoerythrolysis in horse foals and a mule foal: 18 cases (1988-2003). AB - OBJECTIVE: To assess data regarding clinical features, clinicopathologic and blood gas variables, and outcome from horse and mule foals with confirmed neonatal isoerythrolysis (NI). DESIGN: Retrospective case series. ANIMALS: 17 horse and 1 mule foals. PROCEDURE: Medical records of foals (< 14 days old) with NI were reviewed. Information collected included signalment; clinical examination findings; results of hematologic, serum and plasma biochemical, and venous blood gas analyses and urinalysis; treatments; and outcome. RESULTS: Data from 17 horse foals and 1 mule foal with NI (mean age, 71 hours) were evaluated. Many foals had high serum indirect and direct bilirubin concentrations and sorbitol dehydrogenase activity. Whole blood immunoglobulin concentrations were < 400 mg/dL in 4 of 15 foals. Fresh whole blood transfusions were administered to 10 of 18 foals. Among the blood factors implicated in 11 foals, one (Dg) had not previously been associated with NI. Of 10 foals that received blood transfusions, 7 had significant improvements in Hct and hemoglobin concentration and 2 had significant improvements in central venous oxygen tension. Fifteen foals survived to discharge. CONCLUSIONS AND CLINICAL RELEVANCE: Data suggest that blood factor Dg may be associated with NI in foals. Liver disease may be concurrent with NI in foals, and NI can develop in foals with inadequate passive transfer of colostral antibodies. Whole blood transfusions were successful at increasing oxygen carrying capacity and improving peripheral tissue oxygenation in NI-affected foals. With appropriate treatment, the prognosis for foals with NI is good. PMID- 16266018 TI - Comparison of the calving-to-conception interval in dairy cows with different degrees of lameness during the prebreeding postpartum period. AB - OBJECTIVE: To compare calving-to-conception intervals among cows classified as nonlame, moderately lame, or lame during the prebreeding postpartum period and to examine the relationship between severity of lameness and time to conception in cows that were classified as lame. DESIGN: Longitudinal study. ANIMALS: 499 Holstein cows. PROCEDURE: Cows in the prebreeding postpartum period were classified as nonlame, moderately lame, or lame by use of a 6-point locomotion scoring system. Time to conception (days) was compared among cows. A low, medium, or high cumulative locomotion score was assigned to lame cows, and time to conception among those cows was compared. Cows classified as lame were examined on a tilt table for diagnosis and treatment of lameness. RESULTS: 154 (31%), 214 (43%), and 131 (26%) cows were classified as nonlame, moderately lame, and lame, respectively. Most cows classified as lame had laminitis (54%) or disorders of the claw (33%). Median time to conception was 36 to 50 days longer in lame cows than in nonlame cows. Among lame cows, the median time to conception was 66 days longer in cows with high cumulative locomotion scores than in cows with low scores. CONCLUSION AND CLINICAL RELEVANCE: Nonlame cows became pregnant more quickly than lame cows. Lame cows with low cumulative locomotion scores during the prebreeding postpartum period became pregnant sooner than lame cows with high scores. Early diagnosis and intervention may mitigate the effects of lameness and improve reproductive performance in lame dairy cows. PMID- 16266019 TI - Comparison of milk yield in dairy cows with different degrees of lameness. AB - OBJECTIVE: To compare milk yield among cows classified as nonlame, moderately lame, and lame and to examine the relationship between severity of lameness and milk yield in cows classified as lame during the first 100 days after parturition. DESIGN: Longitudinal study. ANIMALS: 465 Holstein cows. PROCEDURE: Cows were examined weekly during the first 100 days after parturition and assigned a lameness score by use of a 6-point locomotion scoring system (ie, 0 to 5). Milk yield was compared among cows classified as nonlame, moderately lame, and lame. Among cows classified as lame (locomotion score > or = 4), milk yield was compared for cows with low, medium, and high cumulative locomotion scores. Cows classified as lame were further examined on a tilt table for diagnosis and treatment of lameness. RESULTS: 84 (18%), 212 (46%), and 169 (36%) cows were classified as nonlame, moderately lame, and lame, respectively. Among cows in their second or later lactations, milk yield in lame cows was significantly lower than that in moderately lame and nonlame cows. In addition, among cows classified as lame, milk yield was significantly lower in cows with high locomotion scores during the first 100 days after parturition, compared with cows with low scores. Most (58%) cows classified as lame had laminitis. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicate a linear relationship between increasing degree of lameness and decreasing milk yield among cows in their second or later lactations. The locomotion scoring system used in this study may be a useful management tool that veterinarians and dairy farmers could adopt for early detection of lameness in dairy cows. PMID- 16266020 TI - Association between hygiene scores and somatic cell scores in dairy cattle. AB - OBJECTIVE: To develop a simple system for scoring hygiene in dairy cattle and determine whether hygiene scores were associated with individual cow somatic cell scores (SCSs). DESIGN: Observational study. ANIMALS: 1,191 cows. PROCEDURE: With the aid of a chart containing line drawings and descriptive text, hygiene scores ranging from 1 (clean) to 5 (dirty) were assigned for 5 body areas: tail head, thigh (lateral aspect), abdomen (ventral aspect), udder, and hind limbs (lower portion). To determine repeatability, hygiene scores were assigned to 75 cows twice by 4 experienced evaluators. To determine accuracy and ease of use, hygiene scores assigned by 14 college students to 23 cows were compared with scores assigned by 2 faculty members. To determine association with SCSs, hygiene scores were assigned to each of 1,093 cows by a single observer. RESULTS: Mean correlation coefficients for hygiene scores assigned twice by 4 experienced evaluators were > or = 0.884, indicating high repeatability. Students indicated that the scoring system was easy to use, and mean correlation coefficient for student and faculty member scores was 0.804. Hygiene scores for the tail head, thigh (lateral aspect), and abdomen (ventral aspect) were not significantly associated with SCS. However, hygiene scores for the udder and hind limbs (lower portion) and udder-hind limb composite scores were significantly associated with SCS, with SCS increasing as scores increased. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that the hygiene scoring system was repeatable, accurate, and easy to use. However, only hygiene scores for the udder and hind limbs and the udder-hind limb composite score were significantly associated with SCS. PMID- 16266022 TI - Wireless: the next IT enabler. PMID- 16266021 TI - Effect of paratuberculosis on culling, milk production, and milk quality in dairy herds. AB - OBJECTIVE: To determine the effect of paratuberculosis on culling, milk production, and milk quality in infected dairy herds. DESIGN: Cross-sectional study. ANIMALS: 689 lactating dairy cows in 9 herds. PROCEDURE: Milk, blood, and fecal samples were obtained from all cows. Fecal samples were evaluated via mycobacterial culture. Serum samples were tested with a commercially available ELISA for antibodies against Mycobacterium avium subsp paratuberculosis, and preserved milk samples were tested with an ELISA for antibodies against M paratuberculosis. Mixed effect and proportional hazards models were used to determine the effect of paratuberculosis on 305-day milk, fat, and protein production; somatic cell count linear score; and the risk of culling. RESULTS: Cows with positive results of bacteriologic culture of feces and milk ELISA produced less milk, fat, and protein, compared with herdmates with negative results. No difference in 305-day milk or fat production was detected in cows with positive results of serum ELISA, compared with seronegative cows. The 3 survival analyses revealed that cows with positive results of each test were at higher risk of being culled than cows with negative results. Paratuberculosis status, as determined by use of all 3 diagnostic tests, was not associated with milk somatic cell count linear score. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that for the 9 herds in this study, paratuberculosis significantly decreased milk production and cow longevity. PMID- 16266023 TI - Sixties soul in the wireless personal computer: go ask Alice. PMID- 16266024 TI - CPOE: the CURE for American healthcare or WMD? PMID- 16266025 TI - Advanced clinician order management a superset of CPOE. PMID- 16266026 TI - Standardized informed consent is a key to improving patient safety. PMID- 16266027 TI - eSignature: the key to healthcare information liquidity. PMID- 16266028 TI - Going, going, gone global. Have you? PMID- 16266029 TI - Voice over IP: how computing technology is being used in mobile communications. AB - This article explains how computing technology was used to address the need for mobile communications among nursing staff. In 2004, nursing staff at Fauquier Hospital relocated from one nursing floor in an older building to two floors in a new structure. This resulted in complaints and supervision issues as nursing managers, who had previously been relatively sedentary, now became quite mobile as they attempted to control nursing operations on two separate floors. Complaints arose from several sources. Nursing staff and managers both complained about the increased difficulty in communicating with each other Physicians expressed frustration to hospital administration at playing "telephone tag" with managers. The solution involved Internet Protocol technology that is in widespread use on most computer networks. The article details how this technology was selected over several other communications technologies and used to implement wireless telephony over the hospital's existing computer network. It reviews key standards and technologies and issues surrounding their use. Finally, the article demonstrates how this computing technology improved patient care by facilitating mobile communications. PMID- 16266030 TI - Experiences incorporating Tablet PCcs into clinical pharmacists' workflow. AB - Tablet PCs are portable computers that combine the power of a laptop with an intuitive pendriven interface that have been heavily promoted for vertical industries such as healthcare. The authors describe their experiences with tablet PCs used by clinical pharmacists in a large academic medical center. A slate tablet with a large screen and wireless networking capability was chosen. Tablet PCs were issued to users with a customized 'pen-friendly" clinical application and secure access to the majority of available electronic patient information. In general, the feedback from the pilot users was positive. Users reported increased efficiency on patient care rounds; they say they reduced or eliminated paper notes and shadow charts from their daily routine. However, researchers also observed that some clinical practice workflow models or clinicians did not benefit from or were hindered by the mobile devices. The department plans to expand the use of tablet PCs through desktop replacement and is developing solutions to increase the applicability of tablet PCs for all clinical models and clinicians. PMID- 16266032 TI - Consumer driven healthcare: strategic, operational, and information technology implications for today's healthcare CIO. AB - This article explores the phenomenon of consumerism in healthcare from an evolutionary perspective and with a view to understanding its implications on the future of our industry. Drawing from the perspectives of leading industry thought leaders and CIOs, it explores the strategic drivers moving our industry toward consumerism and the operational and information technology implications of that trend. By blending real-life examples with potential scenarios, the article is designed to provoke thinking regarding the challenges and opportunities presented by consumerism, thereby informing strategic planning efforts. By doing so, the authors seek to initiate a dialog with readers on this emerging topic while sharing their insights and perspectives with those entrusted with developing consumer-driven healthcare strategies and action plans. PMID- 16266031 TI - Safe use of cellular telephones in hospitals: fundamental principles and case studies. AB - Many industries and individuals have embraced cellular telephones. They provide mobile, synchronous communication, which could hypothetically increase the efficiency and safety of inpatient healthcare. However, reports of early analog cellular telephones interfering with critical life-support machines had led many hospitals to strictly prohibit cellular telephones. A literature search revealed that individual hospitals now are allowing cellular telephone use with various policies to prevent electromagnetic interference with medical devices. The fundamental principles underlying electromagnetic interference are immunity, frequency, modulation technology, distance, and power Electromagnetic interference risk mitigation methods based on these principles have been successfully implemented. In one case study, a minimum distance between cellular telephones and medical devices is maintained, with restrictions in critical areas. In another case study, cellular telephone coverage is augmented to automatically control the power of the cellular telephone. While no uniform safety standard yet exists, cellular telephones can be safely used in hospitals when their use is managed carefully. PMID- 16266033 TI - Hospital implementation of computerized provider order entry systems: results from the 2003 leapfrog group quality and safety survey. AB - A critical element of The Leapfrog Group's strategy for advancing improvements in healthcare is its ongoing survey of hospital patient safety and quality improvement activities, including computerized provider order entry (CPOE) systems. This survey is distinct from other surveys of CPOE adoption because individual hospital responses are publicly disseminated. Furthermore, this survey offers an opportunity to explore the drivers of hospital CPOE adoption before financial incentives for patient safety proliferate, as well as an opportunity to compare the characteristics of participating and non-participating hospitals. Results from the 2003 survey show that only 3.7 percent of the 842 participating hospitals located in The Leapfrog Group's targeted regions had fully implemented a CPOE system consistent with the Leapfrog standard, although 92 percent reported at least planned or partial implementation of a CPOE system. While prior research suggests that a hospital's financial condition should be positively correlated with decisions to invest in CPOE, the analysis generally failed to detect such a relationship. PMID- 16266034 TI - Environments for innovation in healthcare information technology. AB - Our nation's leaders have been aggressively touting the benefits of electronic health records. While the maturation of commercially available products has helped to foster adoption, emerging innovation is evolutionary rather than revolutionary. To create revolutionary healthcare IT, we need environments that allow rapid-cycle, iterative development, and evaluation of innovative healthcare IT in real-world settings. PMID- 16266036 TI - The patient safety institute demonstration project: a model for implementing a local health information infrastructure. AB - The increasing focus on patient safety has uncovered many unsafe conditions in the current US. healthcare system. One of the most glaring problems is the inability of a fragmented healthcare system to provide critical and timely clinical information at the point of care. The Institute of Medicine has called for the development of a National Health Information Infrastructure to rectify this deficiency. This NHII will be built on Local Health Information Infrastructures, or LHIIs. The Patient Safety Institute is a potential model for an LHII that was developed and implemented in Seattle using the Swedish Medical Centers and associated ambulatory clinics. This model was piloted and evaluated among 365 clinical users across three hospitals, three clinics, and family practice residency programs involving access of records of more than 5300 distinct patients within a five-month period and involved the collection of more than 23 million clinical data results. User responses revealed the technology was intuitive to learn, easy to use, easy to navigate, and helpful in clinical care. The PSI demonstration project has developed an approach to the creation and implementation of LHIIs that is potentially transferable to other local communities. PMID- 16266035 TI - Improving physician communication through an automated, integrated sign-out system. AB - Communication failures among physicians are a leading cause of medical errors. The resident sign-out sheet is the primary tool used by house staff to facilitate the sign-out process. The resident sign-out sheet is a structured report, with patient-specific information including demographics, such a patient's name, age, sex, room number, and attending physician; problem list; medications; and allergies. Some physicians use handwritten notes to keep track of this information, while others use freestanding word processor or database programs. In a previous study, the authors described serious inaccuracies in a manually updated word-processor based resident sign-out sheet used by pediatric residents at a tertiary-care children's hospital. An automated and integrated sign-out system (AISS) was subsequently developed that retrieves pertinent patient information from a computerized provider order entry (CPOE) system. The AISS generates a resident sign-out sheet, which includes demographic information, weight, current medications, allergies, and diet orders, as well as optional free text information. The AISS has proven to be enormously popular, increasing physician acceptance of CPOE throughout the organization. This paper discusses lessons learned, including technical, design, and workflow aspects of an integrated resident sign-out sheet. The authors recommend that all future commercial CPOE systems incorporate physician sign-out tools such as the one described in this article. PMID- 16266037 TI - Fast forwarding: the evolution of neurosurgery. The 2005 presidential address. AB - Despite the major social and economic reorganization of medical practice that has taken place during the past 40 years, neurosurgery-the most fascinating specialty in all of clinical medicine-has grown and prospered. Today, this specialty is poised for an era of spectacular advancement and improvement in care; however, significant problems with the potential to retard this growth face neurosurgery. Among these problems is the medical liability situation, which has the potential to destabilize neurosurgical practices and the current health care delivery system. Other issues facing neurosurgery include the potential for loss of the unique nature of the specialty through a conversion to shift-worker surgeons and increasing reliance on profit-seeking institutions for financial stability and liability protection. Lifestyle choices are of growing importance and currently discourage women from entering the field. With a growing knowledge base, there is the recognition that it may not be possible for most individuals to master all aspects of the specialty. There is continued confusion about manpower needs. In addition, some neurosurgeons are choosing to practice in ways that fail to meet the neurosurgeon's obligations to society. There is a growing number of neurosurgeons who dislike providing trauma coverage and there is the potential for some neurosurgeons to give up intracranial neurosurgery. The author believes that it is not competition that will improve the delivery of neurosurgical care and allow for continued growth, but cooperation, and that it will be possible to alleviate many of our problems through increased regionalization of neurosurgical care delivery. This proposal has the potential to promote the formation of neurosurgical teams, ameliorate the problem of physician fatigue, allow greater development of subspecialty skills, and ease the burden of trauma call. It should allow satisfactory solutions to lifestyle considerations and encourage more women to enter the field of neurosurgery. Such a transformation would encourage advances in care to be brought rapidly into the clinical setting and allow neurosurgery to be practiced at the very highest level. PMID- 16266038 TI - Coronary artery disease and carotid endarterectomy. PMID- 16266039 TI - Prediction of coronary artery disease in patients undergoing carotid endarterectomy. AB - OBJECT: The authors determined the factors that predict the coexistence of coronary artery disease (CAD) in patients who undergo carotid endarterectomy (CEA). METHODS: Data from 200 consecutive Japanese patients who underwent CEA for extracranial carotid artery stenosis were studied. Among 73 patients with CAD, 35 (48%) had three-vessel or left main CAD (that is, severe CAD). Peripheral artery disease was an independent predictor of CAD (odds ratio [OR] 2.61, 95% confidence interval [CI] 1.08-6.3). In addition, diabetes mellitus ([DM]; OR 2.8, 95% CI 1.24-6.32) and peripheral artery disease (PAD) (OR 2.83, 95% CI 1.05-7.57) were independent predictors of severe CAD in the 200 patients. The percentage of patients with CAD as well as those with the severe form of the disease increased stepwise as the number of major coronary risk factors in patients increased. Asymptomatic CAD was newly detected during the pre-CEA assessment in 18 (25%) of the 73 patients in whom CAD was eventually diagnosed. Diabetes mellitus was an independent predictor of occult CAD among the 200 patients (OR 4.83, 95% CI 1.53 15.2). CONCLUSIONS: In patients with DM, PAD, or multiple major coronary risk factors who have been scheduled for CEA, one should carefully search for concomitant CAD, especially severe CAD, even when the patient has had no previous episode of angina. PMID- 16266040 TI - Cerebral acid-base homeostasis after severe traumatic brain injury. AB - OBJECT: Brain tissue acidosis is known to mediate neuronal death. Therefore the authors measured the main parameters of cerebral acid-base homeostasis, as well as their interrelations, shortly after severe traumatic brain injury (TBI) in humans. METHODS: Brain tissue pH, PCO2, PO2, and/or lactate were measured in 151 patients with severe head injuries, by using a Neurotrend sensor and/or a microdialysis probe. Monitoring was started as soon as possible after the injury and continued for up to 4 days. During the 1st day following the trauma, the brain tissue pH was significantly lower, compared with later time points, in patients who died or remained in a persistent vegetative state. Six hours after the injury, brain tissue PCO2 was significantly higher in patients with a poor outcome compared with patients with a good outcome. Furthermore, significant elevations in cerebral concentrations of lactate were found during the 1st day after the injury, compared with later time points. These increases in lactate were typically more pronounced in patients with a poor outcome. Similar biochemical changes were observed during later hypoxic events. CONCLUSIONS: Severe human TBI profoundly disturbs cerebral acid-base homeostasis. The observed pH changes persist for the first 24 hours after the trauma. Brain tissue acidosis is associated with increased tissue PCO2 and lactate concentration; these pathobiochemical changes are more severe in patients who remain in a persistent vegetative state or die. Furthermore, increased brain tissue PCO2 (> 60 mm Hg) appears to be a useful clinical indicator of critical cerebral ischemia, especially when accompanied by increased lactate concentrations. PMID- 16266041 TI - Spectrum of subdural fluid collections in spontaneous intracranial hypotension. AB - OBJECT: Spontaneous intracranial hypotension is a noteworthy but commonly misdiagnosed cause of new daily persistent headaches. Subdural fluid collections are frequent radiographic findings, but they can be interpreted as primary rather than secondary pathological entities, and uncertainties exist regarding their optimal management. The authors therefore reviewed their experience with subdural fluid collections in 40 consecutive patients with spontaneous spinal cerebrospinal fluid (CSF) leaks and intracranial hypotension. METHODS: The mean age of the 26 female and 14 male patients was 43 years (range 13-72 years). Subdural fluid collections were present in 20 patients (50%); 12 of these patients (60%) had subdural hygromas alone, and eight (40%) had subacute to chronic subdural hematomas (SDHs) associated with significant mass effect. The subdural hygromas resolved within several days to weeks following treatment of the underlying CSF leak. Three patients with SDHs underwent evacuation of the hematoma prior to the establishment of the diagnosis of spontaneous intracranial hypotension, but the SDHs did not resolve until the underlying spinal CSF leak was treated. In the remaining five patients, the CSF leak was treated primarily and the SDHs resolved over a 1- to 3-month period without the need for evacuation. CONCLUSIONS: Subdural fluid collections are common in spontaneous intracranial hypotension, varying in appearance from thin subdural hygromas to large SDHs associated with significant mass effect. These collections can be safely managed by directing treatment at the underlying CSF leak without the need for hematoma evacuation. PMID- 16266042 TI - Surgical interventions for traumatic lesions of the brachial plexus: a retrospective study of 134 cases. AB - OBJECT: Surgical therapy for traumatic brachial plexus lesions is still a great challenge in the field of peripheral nerve surgery. The aim of this study was to present the results of different surgical interventions in patients with this lesion type. METHODS: One hundred thirty-four patients with traumatic brachial plexus lesions underwent surgery between January 1991 and September 1999. In more than 50% of the patients, injury was caused by a motorbike accident. Patients underwent surgery a mean of 6.3 months posttrauma. The following surgical techniques were applied: neurolysis for nerve lesions in continuity (27 cases), grafting for lesions in discontinuity (149 cases), and neurotization for root avulsions (67 cases). Sixty-five patients were evaluated for at least 30 months (mean follow up 42.1 months) after surgery. Function was graded using the Louisiana State University Health Sciences Center classification system. Only 2% of the patients had Grade 3 or better function preoperatively, increasing to 52% postoperatively. The effect of surgical measures on the functional results for different muscles were compared (supra- or infraspinatus, deltoid, biceps, and triceps muscles); the best results were obtained for biceps muscle function (57% of patients with Medical Research Council Grades M3-M5 function). Graft reconstruction yielded a better outcome than neurotization. Surgery within 5 months posttrauma clearly resulted in improved recovery of motor function compared with later interventions. Sural nerve grafts (monofascicular nerves) showed better results. CONCLUSIONS: The results of neurosurgical interventions for brachial plexus lesions are satisfactory, especially when the operation is performed between 3 and 6 months after trauma. PMID- 16266043 TI - Missile-induced complete lesions of the tibial nerve and tibial division of the sciatic nerve: results of 119 repairs. AB - OBJECT: Very few extensive studies regarding the repair of missile-induced tibial nerve or tibial division complete lesions have been published to date. In this prospective study, the outcomes of such repairs as well as factors influencing them are presented. METHODS: Between 1991 and 1994, 119 patients with missile induced complete lesions of the tibial nerve or tibial division were treated in the neurosurgical department of the Military Medical Academy, Belgrade. After at least 4 years of follow up, the final outcome was defined as poor, insufficient, good, or excellent, based on sensorimotor recovery, electromyoneurography demonstrated recovery, and patient judgment. Good and excellent outcomes were considered to be successful. The influence of the repair level, length of the defect, and preoperative interval on final outcome was also tested. A successful outcome was obtained in 30.3% of high-level, 50% of intermediate-level, and 85.7% of low-level repairs (p < 0.001). On average, the nerve defect and preoperative interval were significantly shorter in patients with a successful outcome. Significant worsening of the outcome was related to a nerve defect longer than 5 cm and a preoperative interval longer than 4 months. Repair level, preoperative interval, and length of the defect were independent predictors of a successful outcome. CONCLUSIONS: A successful outcome is most probable following the low level repairs, within the first 4 months after injury, and using grafts shorter than 5 cm. Other repairs can also be beneficial in preventing dangerous anesthesia of the sole of the foot and enabling almost normal walking. PMID- 16266044 TI - Stereotactic interstitial radiosurgery for cerebral metastases. AB - OBJECT: The Photon Radiosurgery System (PRS) is a miniature x-ray generator that can stereotactically irradiate intracranial tumors by using low-energy photons. Treatment with the PRS typically occurs in conjunction with stereotactic biopsy, thereby providing diagnosis and treatment in one procedure. The authors review the treatment of patients with brain metastases with the aid of the PRS and discuss the indications, advantages, and limitations of this technique. METHODS: Clinical characteristics, treatment parameters, neuroimaging-confirmed outcome, and survival were reviewed in all patients with histologically verified brain metastases who were treated with the PRS at the Massachusetts General Hospital between December 1992 and November 2000. Local control of lesions was defined as either stabilization or diminution in the size of the treated tumor as confirmed by Gd-enhanced magnetic resonance imaging. Between December 1992 and November 2000, 72 intracranial metastatic lesions in 60 patients were treated with the PRS. Primary tumors included lung (33 patients), melanoma (15 patients), renal cell (five patients), breast (two patients), esophageal (two patients), colon (one patient), and Merkle cell (one patient) cancers, and malignant fibrous histiocytoma (one patient). Supratentorial metastases were distributed throughout the cerebrum, with only one cerebellar metastasis. The lesions ranged in diameter from 6 to 40 mm and were treated with a minimal peripheral dose of 16 Gy (range 10-20 Gy). At the last follow-up examination (median 6 months), local disease control had been achieved in 48 (81%) of 59 tumors. An actuarial analysis demonstrated that the survival rates at 6 and 12 months were 63 and 34%, respectively. Patients with a single brain metastasis survived a mean of 11 months. Complications included four patients with postoperative seizures, three with symptomatic cerebral edema, two with hemorrhagic events, and three with symptomatic radiation necrosis requiring surgery. CONCLUSIONS: Stereotactic interstitial radiosurgery performed using the PRS can obtain local control of cerebral metastases at rates that are comparable to those achieved through open resection and external stereotactic radiosurgery. The major advantage of using the PRS is that effective treatment can be accomplished at the time of stereotactic biopsy. PMID- 16266045 TI - Mental health, anxiety, and depression in patients with cerebral aneurysms. AB - OBJECT: Aneurysm disease and its treatment can have an adverse impact on mental health, yet the affects of cerebral aneurysms on general mental health, anxiety, and depression are poorly understood. METHODS: Patients with cerebral aneurysms who were seen at a neurosurgery clinic underwent a structured interview, completed the Hospital Anxiety and Depression Scale and the Medical Outcomes Study 12-item Short Form Health Survey (providing a mental component summary [MCS] score for general mental health), and were assigned functional status scores based on the Glasgow Outcome Scale (GOS), Rankin Scale, and Barthel Index. Rank-order methods were used to assess the relationship between mental health, aneurysm characteristics and history, and functional status. Data were collected in 166 patients (71% women) with a mean age of 53.7 years. Depression was present in 8% of the study population and an anxiety disorder in 17%. Patients with both an unsecured aneurysm and a history of subarachnoid hemorrhage (SAH) tended toward higher anxiety scores (p = 0.086). Higher depression scores were associated with a decreased functional status on the GOS (p = 0.015) and Rankin Scale (p = 0.010). The mean +/- standard deviation adjusted MCS score (37.9 +/- 7.1) was significantly less than that of the US population (p < 0.001). Lower MCS scores were associated with a decreased functional status on the GOS (p = 0.052), Rankin Scale (p < 0.001), and Barthel Index (p = 0.002). CONCLUSIONS: Patients with cerebral aneurysms have increased levels of anxiety and depression and poor general mental health. Those who have experienced an SAH and harbor an unsecured cerebral aneurysm demonstrate increased levels of anxiety. A lower functional status in patients with aneurysms is associated with depression and decreased general mental health. PMID- 16266046 TI - Testing the radiosurgery-based arteriovenous malformation score and the modified Spetzler-Martin grading system to predict radiosurgical outcome. AB - OBJECT: The aim of this study was to validate the radiosurgery-based arteriovenous malformation (AVM) score and the modified Spetzler-Martin grading system to predict radiosurgical outcome. METHODS: One hundred thirty-six patients with brain AVMs were randomly selected. These patients had undergone a linear accelerator radiosurgical procedure at a single center between 1989 and 2000. Patients were divided into four groups according to an AVM score, which was calculated from the lesion volume, lesion location, and patient age (Group 1, AVM score <1; Group 2, AVM score 1-1.49; Group 3, AVM score 1.5-2; and Group 4, AVM score >2). Patients with a Spetzler-Martin Grade III AVM were divided into Grades IIIA (lesion >3 cm) and IIIB (lesion <3 cm). Sixty-two female (45.6%) and 74 male (54.4%) patients with a median age of 37.5 years (mean 37.5 years, range 5-77 years) were followed up for a median of 40 months. The median tumor margin dose was 15 Gy (mean 17.23 Gy, range 15-25 Gy). The proportions of excellent outcomes according to the AVM score were as follows: 91.7% for Group 1, 74.1% for Group 2, 60% for Group 3, and 33.3% for Group 4 (chi-square test, degrees of freedom (df) = 3, p < 0.001). Based on the modified Spetzler-Martin system, Grade I lesions had 88.9% excellent results; Grade II, 69.6%; Grade IIIB, 61.5%; and Grades IIIA and IV, 44.8% (chi-square test, df = 3, p = 0.047). CONCLUSIONS: The radiosurgery based AVM score can be used accurately to predict excellent results following a single radiosurgical treatment for AVM. The modified Spetzler-Martin system can also predict radiosurgical results for AVMs, thus making it possible to use this system while deciding between surgery and radiosurgery. PMID- 16266047 TI - Endovascular management of dissecting vertebrobasilar artery aneurysms in patients presenting with acute subarachnoid hemorrhage. AB - OBJECT: The authors report on a series of 29 patients presenting with acute subarachnoid hemorrhage (SAH) related to the rupture of a vertebrobasilar dissecting aneurysm. Special attention was focused on embolization techniques and immediate and midterm anatomical and clinical outcomes. METHODS: Between March 1994 and January 2003, 29 patients presented with acute SAH caused by the rupture of a vertebrobasilar dissecting aneurysm. Eleven patients (37.9%) had Hunt and Hess Grade I SAH, four (13.8%) Grade II, six (20.7%) Grade III, five (17.2%) Grade IV, and three (10.3%) Grade V. Aneurysms were classified into five groups based on lesion location, and treatment courses were decided. All patients except two were treated by endovascular trapping of the aneurysm with concomitant occlusion of the involved vertebral artery (VA). No technical or clinical complication was observed in 28 patients (97%). Aneurysm perforation occurred during the procedure in one patient (3%). There was evidence of aneurysm recanalization in one patient. One patient with Hunt and Hess Grade IV SAH and two patients with Grade V SAH died. One patient died of respiratory infection 1 year after aneurysm trapping. One patient presented with a recurrent hemorrhage 1 month after treatment and died. Overall morbidity and mortality rates were 13.8 and 17.2%, respectively. CONCLUSIONS: Twenty-nine patients with acute SAH due to rupturing of vertebrobasilar dissecting aneurysms were treated using endovascular techniques. In most cases, endovascular trapping of the aneurysm and concomitant occlusion of the VA was technically and clinically successful. PMID- 16266048 TI - Limitations of three-dimensional reconstructed computerized tomography angiography after clip placement for intracranial aneurysms. AB - OBJECT: The authors compared the usefulness of three-dimensional (3D) reconstructed computerized tomography (CT) angiography with 3D digital subtraction (DS) angiography in assessing intracranial aneurysms after clip placement. A retrospective review of clinical cases was performed. METHODS: Between May 2001 and May 2003, 17 patients with a total of 20 intracranial aneurysms underwent 3D CT and 3D DS angiography following clip placement. The authors assessed the presence or absence of residual aneurysm necks and stenoocclusive changes in the parent artery and the neighboring artery. The efficacy of CT angiographic visualization was also evaluated. In 12 of the 20 aneurysms, both 3D modalities similarly demonstrated the residual aneurysm neck and stenoocclusive changes in the parent artery and neighboring artery. Three dimensional CT angiography failed to demonstrate three of the aneurysms, and the studies were not considered suitable for evaluation because of the presence of metallic artifacts. In the remaining five studies, the 3D CT angiograms did not effectively demonstrate the neighboring and parent arteries. The detectability of residual aneurysm necks was correlated with the clip material and with the number of clips applied. CONCLUSIONS: Three-dimensional DS angiography is still necessary in cases involving multiple clips or with cobalt alloy clips because the clips appear as metal artifacts on 3D CT angiography. PMID- 16266049 TI - A proposed parent vessel geometry-based categorization of saccular intracranial aneurysms: computational flow dynamics analysis of the risk factors for lesion rupture. AB - OBJECT: The authors created a simple, broadly applicable classification of saccular intracranial aneurysms into three categories: sidewall (SW), sidewall with branching vessel (SWBV), and endwall (EW) according to the angiographically documented patterns of their parent arteries. Using computational flow dynamics analysis (CFDA) of simple models representing the three aneurysm categories, the authors analyzed geometry-related risk factors such as neck width, parent artery curvature, and angulation of the branching vessels. METHODS: The authors performed CFDAs of 68 aneurysmal geometric formations documented on angiograms that had been obtained in patients with 45 ruptured and 23 unruptured lesions. In successfully studied CFDA cases, the wall shear stress, blood velocity, and pressure maps were examined and correlated with aneurysm rupture points. Statistical analysis of the cases involving aneurysm rupture revealed a statistically significant correlation between aneurysm depth and both neck size (p < 0.0001) and caliber of draining arteries (p < 0.0001). Wider-necked aneurysms or those with wider-caliber draining vessels were found to be high-flow lesions that tended to rupture at larger sizes. Smaller-necked aneurysms or those with smaller-caliber draining vessels were found to be low-flow lesions that tended to rupture at smaller sizes. The incidence of ruptured aneurysms with an aspect ratio (depth/neck) exceeding 1.6 was 100% in the SW and SWBV categories, whereas the incidence was only 28.75% for the EW aneurysms. CONCLUSIONS: The application of standardized categories enables the comparison of results for various aneurysms' geometric formations, thus assisting in their management. The proposed classification system may provide a promising means of understanding the natural history of saccular intracranial aneurysms. PMID- 16266050 TI - Tenascin-C-coated platinum coils for acceleration of organization of cavities and reduction of lumen size in a rat aneurysm model. AB - OBJECT: Detachable platinum coils are widely used in the endovascular treatment of intracranial aneurysms. The use of coil placement produces a higher incidence of aneurysm recurrence compared with surgical clipping. To reduce the incidence of recurrence by promoting clot organization, the authors designed a platinum coil coated with tenascin-C (TNC), an extracellular matrix glycoprotein, and then histologically examined tissue responses. METHODS: Platinum coils were prepared by successive coatings with cationic polyethyleneimine and anionic heparin and then TNC or basic fibroblast growth factor (bFGF) was immobilized by affinity binding to the heparin. Six unmodified, six heparin-coated, six bFGF-coated, or eight TNC-coated platinum coils were inserted into ligated common carotid arteries (CCAs) of adult male rats, and CCA segments were harvested after 14 or 28 days. The percentages of organized areas occupying the luminal cavity in unmodified, heparin-coated, bFGF-coated, and TNC-coated groups were 4.8 +/- 4.6, 1.6 +/- 1.1, 17.9 +/- 10.7, and 93.4 +/- 6.9%, respectively. In addition, the mean lumen size in the TNC-coated group (0.35 +/- 0.23 mm2) was reduced to less than half that of the unmodified group (0.72 +/- 0.21 mm2). Immunohistochemical analysis revealed that alpha-smooth muscle actin-positive cells were a major cellular component of the organized tissue within the TNC-coated coils but not in the bFGF group. Collagen fibrils in the organized areas were also much thicker and denser with TNC-coated coils than with bFGF-coated coils. CONCLUSIONS: Placement of TNC-coated coils can remarkably accelerate organization of luminal cavities and reduce their volume, providing improved efficacy of these coils for endovascular embolization. PMID- 16266051 TI - Influence of age on stroke outcome following transient focal ischemia. AB - OBJECT: More than 100 clinical trials based on animal models have failed to identify a clinically effective neuroprotectant for stroke. Current models of stroke do not account adequately for aging nor do they incorporate the use of female animals. The authors evaluated the pathological and physiological differences in stroke in young, adult, and elderly female rats. METHODS: Three groups of female Sprague-Dawley rats were studied. Nine rats were divided into three groups: young (3 months); adult (9 months); and elderly (18 months). Intraluminal filament occlusion was performed for 120 minutes while cerebral blood flow was monitored. Physiological parameters were assessed. Infarction volumes were quantified at 24 hours. The mean arterial pressure increased in the young animals (103 +/- 3.51 mm Hg; p < 0.001) during occlusion and decreased in the elderly group (65.56 +/- 3.03 mm Hg; p < 0.01). Cortical and striatal infarction volumes in the elderly animals were substantially larger (p < 0.05). Young animals exhibited a lesser decrement in cerebral blood flow (p < 0.05) during ischemia. CONCLUSIONS: This study reinforces the importance of using older animals for the researching and treatment of stroke. Elderly animals show differences in response mechanisms, ischemic consequences, and histological changes. These differences may partially explain the current lack of success involved in using young-animal models to predict the clinical efficacy of neuroprotective agents. PMID- 16266052 TI - Effect of atorvastatin on spatial memory, neuronal survival, and vascular density in female rats after traumatic brain injury. AB - OBJECT: Atorvastatin administered after traumatic brain injury (TBI) induced by controlled cortical impact promotes functional improvement in male rats. Note, however, that parallel studies have not been performed in female rats. Therefore, the authors tested the effect of atorvastatin on TBI in female rats. METHODS: Atorvastatin (1 mg/kg/day) was orally administered for 7 consecutive days in female Wistar rats starting I day after TBI; control animals received saline. Modified neurological severity scores, the corner turn test, and the Morris water maze test were used to evaluate functional response to treatment. Rats were killed on Day 15 post-TBI, and brain tissue samples were processed for immunohistochemical staining. Atorvastatin administration after brain injury significantly promoted the restoration of spatial memory but did not reduce sensorimotor functional deficits. Treatment of TBI with atorvastatin increased neuronal survival in the CA3 region and the lesion boundary zone and prevented the loss of neuronal processes of damaged neurons in the hippocampal CA3 region but not in the lesion boundary zone on Day 15 after TBI. The protective effect of atorvastatin on the injured neurons perhaps is mediated by increasing the density of vessels in the lesion boundary zone and the hippocampus after TBI. CONCLUSIONS: . These data indicate that atorvastatin is beneficial in the treatment of TBI in female rats, although the effect may differ between sexes. PMID- 16266053 TI - Gliomatosis cerebri: quantitative proof of vessel recruitment by cooptation instead of angiogenesis. AB - OBJECT: Gliomas are the most common primary brain tumors, many of which (especially astrocytic and oligodendroglial neoplasms) are characterized by diffuse infiltrative growth in the preexisting brain tissue. Gliomatosis cerebri is a rare glial tumor and represents an extreme example of such diffuse infiltrative growth. This growth pattern not only hampers curative treatment but also allows for vessel cooptation rather than tumor angiogenesis as a way of vessel recruitment by the tumor tissue. The goal of this study was to establish the extent to which tumor angiogenesis occurs in gliomatosis cerebri. METHODS: Computerized image analysis was performed to assess quantitatively two microvascular parameters (vessel density and diameter) in different areas of a brain harboring a gliomatosis cerebri. These regions were the cerebral white and gray matter in which there was a diffuse infiltrative tumor, cerebral white and gray matter in which there was a more compact growth pattern of tumor cells, and normal cerebral white and gray matter. In addition, the authors performed immunohistochemical stainings for blood-brain barrier (BBB) characteristics (Glut 1 and PgP) on samples obtained in these different areas. The results of the quantitative analysis strongly indicated that in gliomatosis cerebri tumor, angiogenesis was completely absent, a finding that is corroborated by the fact that the microvasculature in gliomatosis cerebri persists in exhibiting immunohistochemical characteristics of the BBB. CONCLUSIONS: The results of this study may help resolve the difficulties in radiological detection and delineation of the diffuse infiltrative part of glial brain tumors and put the expectations for antiangiogenic treatment of such tumors into perspective. PMID- 16266054 TI - Secondary ischemia impairing the restoration of ion homeostasis following traumatic brain injury. AB - OBJECT: It is well established that posttraumatic secondary ischemia contributes to poor outcome. Ion dysfunction leading to cytotoxic edema is a primary force in the formation of ischemic brain edema and is a principal component of traumatic brain swelling. Because cell swelling is the result of net ion and water movement, it is crucial to have a thorough understanding of these transient phenomena. The purpose of this study was to characterize the effects of secondary ischemia following traumatic brain injury (TBI) on the ability to restore ion homeostasis. METHODS: Twenty-four Sprague-Dawley rats were divided into four groups of six animals each. The rats underwent transient forebrain ischemia via bilateral carotid artery occlusion combined with hypotension: 15 minutes of forebrain ischemia (Group 1); 60 minutes of forebrain ischemia (Group 2); impact acceleration/TBI (Group 3); and impact acceleration/TBI followed by 15 minutes of ischemia (Group 4). Ischemia resulted in a rapid accumulation of [K+]e:41.94 +/- 13.65 and 66.33 +/- 6.63 mM, respectively, in Groups 1 and 2, with a concomitant decrease of [Na+]e:64 +/- 18 mM and 72 +/- 11 mM in Groups 1 and 2. Traumatic brain injury resulted in a less severe although identical trend in ion dysfunction ([K+]e 30.42 +/- 11.67 mM and [Na+]e 63 +/- 33 mM). Secondary ischemia resulted in prolonged and sustained ion dysfunction with a concomitant elevation of intracranial pressure (ICP). CONCLUSIONS: Analysis of these results indicates that ischemia and TBI are sublethal in isolation; however, when TBI is associated with secondary ischemia, ion dysfunction is sustained and is associated with elevated ICP. PMID- 16266056 TI - Effect of perfluorocarbons on brain oxygenation and ischemic damage in an acute subdural hematoma model in rats. AB - OBJECT: This study was conducted to determine whether perfluorocarbons (PFCs) improve brain oxygenation and reduce ischemic brain damage in an acute subdural hematoma (SDH) model in rats. METHODS: Forty adult male Sprague-Dawley rats were allocated to four groups: (1) controls, acute SDH treated with saline and 30% O2; (2) 30-PFC group, acute SDH treated with PFC infusion in 30% O2; (3) 100-O2 group, acute SDH treated with 100% O2; and (4) 100-PFC group, acute SDH treated with PFC plus 100% O2. Ten minutes after the induction of acute SDH, a single dose of PFC was infused and 30% or 100% O2 was administered simultaneously. Four hours later, half of the rats were killed by perfusion for histological study to assess the extent of ischemic brain damage. The other half were used to measure brain tissue oxygen tension (PO2). The volume of ischemic brain damage was 162.4 +/- 7.6 mm3 in controls, 165.3 +/- 11.3 mm3 in the 30-PFC group, 153.4 +/- 17.3 mm3 in the 100-O2 group, and 95.9 +/- 12.8 mm3 in the 100-PFC group (41% reduction compared with controls, p = 0.002). Baseline brain tissue PO2 values were approximately 20 mm Hg, and after induction of acute SDH, PO2 rapidly decreased and remained at 1 to 2 mm Hg. Treatment with either PFC or 100% O2 improved brain tissue PO2, with final values of 5.14 and 7.02 mm Hg, respectively. Infusion of PFC with 100% O2 improved brain tissue PO2 the most, with a final value of 15.16 mm Hg. CONCLUSIONS: Data from the current study demonstrated that PFC infusion along with 100% O2 can significantly improve brain oxygenation and reduce ischemic brain damage in acute SDH. PMID- 16266055 TI - Differences in infarct evolution between lipopolysaccharide-induced tolerant and nontolerant conditions to focal cerebral ischemia. AB - OBJECT: Although brain tissue may be protected by previous preconditioning, the temporal evolution of infarcts in such preconditioned brain tissue during focal cerebral ischemia is largely unknown. Therefore, in this study the authors engaged in long-term observation with magnetic resonance (MR) imaging to clarify the difference in lesion evolution between tolerant and nontolerant conditions. METHODS: Bacterial lipopolysaccharide (LPS; 0.9 mg/kg) was administered intravenously to induce cross-ischemic tolerance. Focal cerebral ischemia was induced 72 hours later in spontaneously hypertensive rats. Serial brain MR images were obtained 6 hours, 24 hours, 4 days, 7 days, and 14 days after ischemia by using a 7.05-tesla unit. Lesion-reducing effects were evident 6 hours after ischemia in the LPS group. Preconditioning with LPS does not merely delay but prevents ischemic cell death by reducing lesion size. Lesion reduction was a sustained effect noted up to 14 days after ischemia. Reduction of local cerebral blood flow (ICBF) in the periinfarct area was significantly inhibited in the LPS group, which was correlated with endothelial nitric oxide synthase (eNOS) expression. CONCLUSIONS: Significant preservation of ICBF in the periinfarct area, which is relevant to sustained upregulation of eNOS, could be a candidate for the long-term inhibiting effect on infarct evolution in the LPS-induced tolerant state. PMID- 16266057 TI - In vitro and in vivo effects of probucol on hydrolysis of asymmetric dimethyl L arginine and vasospasm in primates. AB - OBJECT: Increased cerebrospinal fluid (CSF) levels of asymmetric dimethyl L arginine (ADMA), an endogenous inhibitor of endothelial nitric oxide synthase (eNOS), are associated with delayed vasospasm after subarachnoid hemorrhage (SAH); however, the source, cellular mechanisms, and pharmacological inhibition of ADMA production following SAH are unknown. METHODS: In an in vitro experiment involving human umbilical vein endothelial cells (HUVECs), the authors examined mechanisms potentially responsible for increased ADMA levels during vasospasm and investigated whether this increase can be inhibited pharmacologically. In a second study, an in vivo experiment, the authors used probucol, which effectively inhibited ADMA increase in HUVEC cultures in vitro, in a randomized double-blind placebo-controlled experiment in a primate model of delayed cerebral vasospasm after SAH. Oxidized low-density lipids (OxLDLs; positive control; p < 0.02) and bilirubin oxidation products (BOXes; p < 0.01), but not oxyhemoglobin (p = 0.74), increased ADMA levels in HUVECs. Probucol inhibited changes in ADMA levels evoked by either OxLDLs (p < 0.001) or BOXes (p < 0.01). Comparable changes were observed in cell lysates. In vivo probucol (100 mg/kg by mouth daily) did not alter serum ADMA levels on Days 7, 14, and 21 after SAH compared with levels before SAH, and these levels were not different from those observed in the placebo group (p = 0.3). Despite achieving therapeutic levels in plasma and measurable levels in CSF, probucol neither prevented increased CSF ADMA levels nor the development of vasospasm after SAH. Increased CSF ADMA and decreased nitrite levels in both groups were strongly associated with the degree of delayed vasospasm after SAH (correlation coefficient [CC] 0.5, 95% confidence interval [CI] 0.19-0.72, p < 0.002 and CC -0.43, 95% CI -0.7 to -0.05, p < 0.03, respectively). CONCLUSIONS: Bilirubin oxidation products, but not oxyhemoglobin, increased ADMA levels in the HUVEC. Despite its in vitro ability to lower ADMA levels, probucol failed to inhibit increased CSF ADMA and decreased nitrite levels, and it did not prevent delayed vasospasm in a primate SAH model. PMID- 16266058 TI - A triple bifurcation aneurysm model for evaluating complex endovascular therapies in dogs. AB - OBJECT: Animal aneurysm models are required for the study of the hemodynamics and pathophysiology of intracranial aneurysms in humans and so that experimental treatments can be tested prior to clinical trials. The authors developed a canine model that consistently produces up to three bifurcation aneurysms similar in morphological features and hemodynamics to human intracranial aneurysms. METHODS: In 10 mongrel dogs, a harvested segment of the external jugular vein was anastamosed to an external carotid artery (CA)-lingual artery bifurcation arteriotomy site to create a lateral bifurcation aneurysm. The surgery was repeated on the contralateral side in each animal to form a second lateral bifurcation aneurysm and, in five dogs, a CA-CA crossover anastomosis was also performed to create a terminal bifurcation aneurysm. Nineteen of 20 lateral bifurcation aneurysms were confirmed in 10 dogs by diagnostic angiography 7 to 14 days after surgery. Aneurysm fundus-to-neck ratios ranged from 1 to 2, depending on the size of the arteriotomy. The terminal bifurcation aneurysms were confirmed in all five dogs by diagnostic angiography 7 to 14 days after the procedure. The authors later tested endovascular techniques for embolizing the aneurysms. CONCLUSIONS: Three bifurcation aneurysms of sufficient size for endovascular access can be created in a reproducible fashion in the same animal. This model is useful for studying complex endovascular procedures in aneurysms that mimic the human condition and for testing new devices and techniques. PMID- 16266059 TI - Controlled release of a nitric oxide donor for the prevention of delayed cerebral vasospasm following experimental subarachnoid hemorrhage in nonhuman primates. AB - OBJECT: Results of prior studies in rats and rabbits show that the alteration of vasomotor tone in vasospasm following periadventitial blood exposure may be reversed, at least in part, by the administration of compounds releasing nitric oxide (NO). The authors have now generalized this finding to nonhuman primates. METHODS: Ten cynomolgus monkeys underwent cerebral angiography before and 7 days following the induction of subarachnoid hemorrhage (SAH) by the placement of 2 to 3 ml clotted autologous blood around the supraclinoid carotid, proximal anterior cerebral, and proximal middle cerebral arteries. An ethylene vinyl acetate copolymer, either blank (five animals) or containing 20% w/w (Z)-1-[2-(2 aminoethyl)-N-(2-aminoethyl)amino]diazen-1-ium-1,2-diolate (DETA/NO, 4.3 mg/kg; five animals) was placed adjacent to the vessels at the time of surgery. Animals were killed on Day 7 post-SAH following repeated cerebral angiography. The mean percentage of control vascular areal fraction was calculated from angiograms. Cerebral vessels were sectioned and the mean percentage of lumen patency was calculated. One animal that had received the DETA/NO polymer died prior to repeated angiography. In the remaining animals, DETA/NO caused a significant decrease in vasospasm compared with controls, according to both angiographic (84.8 +/- 8.6 compared with 56.6 +/- 5.2%, respectively, p < 0.05) and histological studies (internal carotid artery 99.3 +/- 1.8 compared with 60.1 +/- 4.4%, respectively, p < 0.001; middle cerebral artery 98.4 +/- 3 compared with 56.1 +/- 3.7%, respectively, p < 0.001; and anterior cerebral artery 89.2 +/- 8.5 compared with 55.8 +/- 6.3%, respectively, p < 0.05). CONCLUSIONS: The controlled release of DETA/NO is effective in preventing delayed cerebral vasospasm in an SAH model in nonhuman primates. The death of one animal in the treatment group indicates that the present dosage is at the threshold between therapeutic efficacy and toxicity. PMID- 16266060 TI - Acute obstructive hydrocephalus associated with infratentorial subdural hygromas complicating Chiari malformation Type I decompression. Report of two cases and literature review. AB - Obstructive hydrocephalus complicating foramen magnum decompression (FMD) for a Chiari malformation (CM) Type I is rare. Two female patients (17 and 55 years old) presented with strain-related headaches. In both cases magnetic resonance (MR) imaging studies confirmed a CM Type I, which was accompanied by syringomyelia in one case. Both patients underwent uncomplicated FMD with good initial recovery. Unfortunately, conditions in both patients deteriorated, with severe headaches occurring between Days 5 and 6 post-FMD. Decreased consciousness occurred in one case. In both patients, computerized tomography scanning demonstrated an acute obstructive hydrocephalus associated with bilateral infratentorial extraaxial fluid collections (EAFCs). In addition, left parafalcine and convexity EAFCs were present in one case. An emergency external ventricular drain was required in one patient, with delayed conversion to a ventriculo-peritoneal shunt. Spontaneous resolution occurred in the other patient without neurosurgical intervention. In both cases, MR imaging confirmed that each EAFC was subdural, resembled cerebrospinal fluid (CSF), and had distorted the superior cerebellum anteroinferiorly. Despite upper fourth ventricle/aqueduct compromise in one case, normal aqueduct flow artifacts were apparent on examination. All EAFCs resolved spontaneously. Obstructive hydrocephalus complicating FMD is rare but invariably associated with infratentorial EAFCs, which were confirmed to be subdural hygromas in this report. The authors assert that hydrocephalus results from upper fourth ventricle/aqueduct compromise as a result of CSF subdural dissection following a pinhole arachnoid tear on durotomy. Because such hygromas spontaneously resolve, permanent shunt insertion should be avoided. PMID- 16266061 TI - Transvenous embolization for a purely intraorbital arteriovenous fistula. Case report. AB - Purely intraorbital arteriovenous fistulas (AVFs) are rare, and their clinical management is controversial. The authors successfully treated a patient with an intraorbital AVF by transvenous embolization alone. An accurate distinction between an arteriovenous malformation (AVM), which is characterized by the existence of a nidus, and an AVF, which has no nidus, is important and requires superselective ophthalmic artery angiography. Treatment of an intraorbital AVF by transvenous embolization can improve visual function. PMID- 16266062 TI - Extended remission of a recurrent median nerve malignant peripheral nerve sheath tumor after multimodal treatment. Case report. AB - Malignant peripheral nerve sheath tumors (MPNSTs) are difficult to control despite aggressive treatment. In this report the authors describe the treatment and follow-up review of a patient with neurofibromatosis Type I who harbored a recurrent median nerve MPNST. The man underwent preoperative intraarterial and intravenous chemotherapy followed by additional surgery for gross-total removal and postoperative radiotherapy. Two courses of preoperative intraarterial cisplatin and intravenous Adriamycin produced significant tumor shrinkage. Gross total removal of the remaining tumor without amputation of the arm was followed by fractionated radiotherapy (total minimum tumor dose 6485 cGy, maximal dose 6575 cGy). The patient is alive 9.5 years after treatment without evidence of tumor recurrence and with only focal median nerve functional deficits. A review of the patient's treatment is warranted to provide a description of a regimen that may be useful in the treatment of similar patients in the future. PMID- 16266063 TI - CSF pulse pressure and B waves. PMID- 16266064 TI - Temporal lobe epilepsy. PMID- 16266065 TI - Single-level lumbar fusion in chronic discogenic low-back pain: psychological and emotional status as a predictor of outcome measured using the 36-item Short Form. AB - OBJECT: The authors examined the effect of psychological and emotional status on the outcome in patients with suspected chronic discogenic low-back pain (LBP) who have undergone lumbar fusion. METHODS: The authors retrospectively analyzed the medical records, including the results of the 36-item Short Form (SF-36), of 57 consecutive patients (mean age 42.7 years) who underwent single-level lumbar reconstructive surgery between 1994 and 2000. The SF-36 physical component summary (PCS) and mental component summary (MCS) domains were evaluated. Data were sorted into the following categories: excellent, good, fair, same, and worse. Scores greater than 40 for MCS and PCS were defined as "normal" according to US general population data provided by the Medical Outcomes Trust. Of 57 patients, 47 completed postoperative SF-36 surveys at 1 year and 36 completed the 2-year follow-up surveys. Analysis showed that preoperative MCS scores exhibited a significant, direct correlation with PCS score improvements at 1 (r = 0.584, p = 0.000) and 2 (r = 0.623, p = 0.000) years after surgery. In patients in whom preoperative MCS scores reflected normal status, outcomes were excellent or good in 60% at 1-year (18 of 30 cases) and 2-year (15 of 25 cases) follow-up intervals. Patients in whom MCS scores represented abnormal status had less satisfactory outcomes, with excellent or good outcome in only two (18.2%) of all patients at the 2-year follow-up study. CONCLUSIONS: Analysis of the data suggests that psychological and emotional distress may negatively affect postoperative outcome in patients with chronic discogenic LBP. The SF-36 may be easily and effectively used to measure both preoperative psychosocial distress and postoperative outcome. PMID- 16266066 TI - Spinal stenosis due to ossified lumbar lesions. AB - OBJECT: Spinal stenosis due to lumbar ossified lesions is a rare pathological entity. The authors retrospectively evaluated the clinical features and surgical results associated with cases involving lumbar ossified lesion-induced stenosis. METHODS: Data obtained in 20 surgically treated patients with lumbar hyperostotic spinal stenosis were included. To evaluate the background of the disease, body mass index and general complications were assessed. Whole-spine radiological examination was conducted. The presence of ossification of the posterior longitudinal ligament or ossification of the ligamentum flavum was evaluated. Surgical results were classified according to the Japanese Orthopaedic Association (JOA) scale. In the patients in whom neurological deterioration was observed during follow up, the causes of deterioration were reviewed. Seven patients (35%) were obese and six patients (30%) suffered diabetes mellitus. Twelve patients harbored coexisting cervical and/or thoracic ossified lesions. The overall mean JOA score improved from 10.2 to a peak of 22.5; at last follow up examination the mean JOA score was 20.9. In female and older patients with a long history of preoperative symptoms, a low preoperative JOA score, and other spinal lesions, recovery tended to be poorer. Recovery was poor in one patient, and neurological deterioration due to coexisting ossified spinal lesions occurred in another patient during the follow-up period. CONCLUSIONS: Because coexisting ossified lesions were frequently seen, whole-spine analysis is recommended. Early diagnosis and appropriate treatment are important to achieve a better surgical outcome. PMID- 16266067 TI - Coralline hydroxyapatite and laminectomy-derived bone as adjuvant graft material for lumbar posterolateral fusion. AB - OBJECT: The purpose of this study was to evaluate the effectiveness of coralline hydroxyapatite (CHA) and laminectomy-derived bone as an adjuvant graft material when combined with autogenous iliac bone graft (AIBG) in posterolateral fusion (PLF). METHODS: This prospective, case-control study involved 58 patients who underwent lumbar instrumentation-augmented PLF for degenerative spinal stenosis induced segmental instability between July 2000 and June 2001. The patients were divided into three groups. Laminectomy bone and AIBG were placed in the right intertransverse process space in Group 1 (20 patients), CHA and AIBG were placed in Group 2 (19 patients), and laminectomy bone and CHA were placed in Group 3 (19 patients). Pure autogenous iliac cancellous bone graft was placed in the left intertransverse process space in all three groups of patients. Successful fusion was determined by two spine surgeons after examining the plain, anteroposterior, bilateral oblique, and lateral flexion-extension radiographs. If the examiners did not agree on fusion status, fine-cut computerized tomography scans of the fusion mass were used to make the final decision. The chi-square test was used to compare the fusion rate at different time intervals among the three groups. CONCLUSIONS: Pure AIBG placed in left intertransverse process space was associated with the best fusion rate. After 6 months, CHA produced a comparable result to laminectomy-derived bone when combined with AIBG. When laminectomy bone was mixed with CHA, the combination failed to yield a satisfactory fusion rate (57.9%) even 1 year after surgery if no AIBG was added. PMID- 16266068 TI - The use of closed-suction irrigation systems to manage spinal infections. AB - OBJECT: Management of infection in the hardware system-fixated spine has proven to be problematic. In many cases, instrumentation is required to provide stability or to maintain correction of deformity, and removal could be hazardous. The authors describe the use of closed continuous irrigation to treat spinal wound infections in patients with fixation systems; irrigation can be used in all parts of the spine with excellent results. METHODS: The authors conducted a retrospective chart-based review of cases in which spinal instrumentation procedures were performed consecutively during a 10-year period. Infection developed in 36 patients. The infections involved the cervical, thoracic or thoracolumbar, and lumbar regions. Anterior and posterior drains were placed in one patient. In one patient refractory infections responded to replacement of the dual-inflow port drainage system. In all cases the wound infection completely resolved. There were no cases that required the removal of the fusion mass or instrumentation. In all cases progression to solid fusion was achieved. CONCLUSIONS: Closed irrigation systems can be used effectively to manage anterior or posterior cervical, thoracic, and lumbar wound infections. These systems preclude the explantation of the instrumentation and allow spinal stability to be preserved. The authors noted no evidence of recurrent infection or failed fusion. PMID- 16266069 TI - Clinical and radiological results after parapedicular screw fixation of the thoracic spine. AB - OBJECT: The proximity of thoracic pedicles to the spinal cord and neurovascular structures has prompted numerous efforts to obtain proper screw placement. The aim of this study was to evaluate the feasibility and efficacy of thoracic parapedicular (intercostovertebral) screw fixation by following anatomical landmarks and using fluoroscopy. METHODS: Thoracic parapedicular screw fixation was performed in 41 patients (328 screws). Postoperative computerized tomography scans were obtained in all cases to analyze the position of each screw with respect to the pedicle rib unit, the spinal canal, and the vertebral body (VB) according to a proposed novel grading system. Additionally, screw convergence angles were calculated for each instrumented level. There was no additional morbidity associated with parapedicular screw insertion. In a single case a screw suspected of entering the spinal canal was revised without causing neurological damage. The remaining screws were placed entirely within the pedicle rib unit and VB without breaching the neural foramina, pleura, or vascular structures. CONCLUSIONS: Thoracic and thoracolumbar instability can be treated adequately and safely by using parapedicular screw fixation guided by anatomical landmarks and fluoroscopic imaging. The use of computer-aided navigation may be beneficial but does not appear to be mandatory. PMID- 16266070 TI - Stereotactic radiosurgery for spinal metastases from renal cell carcinoma. AB - OBJECT: The role of stereotactic radiosurgery in treating renal cell carcinoma (RCC) metastases to the spine has previously been limited. In this study the authors evaluated the clinical outcome in patients with spinal RCC who underwent single-fraction radiosurgery. METHODS: Forty-eight patients with 60 RCC metastases to the spine (six cervical, 26 thoracic, 18 lumbar, and 10 sacral) were treated with a single-fraction radiosurgery technique and were followed for a period of 14 to 48 months (median 37 months). All patients were successfully treated in an outpatient setting. The tumor volume ranged from 5.5 to 203 cm3 (mean 61.9 cm3). Forty-two of the total 60 lesions had been previously treated with external-beam radiation therapy (EBRT). The maximum tumor dose was maintained at 17.5 to 25 Gy (mean 20 Gy). The volume of the spinal cord exposed to greater than 8 Gy ranged from 0.01 to 3 cm3 (mean 0.64 cm3); the volume of the spinal canal at the cauda equina level exposed to greater than 8 Gy ranged from 0.01 to 2.2 cm3 (mean 0.65 cm3). No radiation-induced toxicity occurred during the follow-up period. Axial and radicular pain improved in 34 (89%) of 38 patients who were treated primarily for pain. Tumor control was demonstrated in seven of eight patients treated primarily for radiographically documented tumor progression. In time six patients required open surgical intervention for tumor progression that had caused neurological dysfunction after radiosurgery. CONCLUSIONS: Spinal radiosurgery can be a successful therapeutic modality for the delivery of large-dose single-fraction radiation to RCC spinal metastases that are often poorly controlled with conventional EBRT modalities. PMID- 16266071 TI - Combination kyphoplasty and spinal radiosurgery: a new treatment paradigm for pathological fractures. AB - OBJECT: Patients with symptomatic pathological compression fractures require spinal stabilization surgery for mechanical back pain control and irradiation for the underlying malignant process. The authors evaluated a treatment paradigm of closed fracture reduction and fixation involving kyphoplasty and subsequent spinal radiosurgery. METHODS: Twenty-six patients (six men and 20 women, mean age 72 years) with pathological compression fractures (16 thoracic and 10 lumbar) were prospectively evaluated. Histological diagnoses included 11 lung, nine breast, four renal, one cholangiocarcioma, and one ocular melanoma. Seven lesions had received prior external-beam radiation therapy. All patients underwent kyphoplasty that involved the percutaneous transpedicular technique. Fiducial markers allowing for image guidance during CyberKnife treatment were placed, at time of the kyphoplasty, in the pedicles at adjacent levels. Patients underwent single-fraction radiosurgery (mean time after kyphoplasty 12 days) in an outpatient setting. The tumor dose was maintained at 16 to 20 Gy (mean 18 Gy) to the 80% isodose line. The treated tumor volume ranged from 12.7 to 37.1 cm3. No acute radiation-induced toxicity or new neurological deficit occurred during the follow-up period (range 11-24 months, median 16 months). Axial pain improved in 24 (92%) of 26 patients. CONCLUSIONS: The combined kyphoplasty and spinal radiosurgery treatment paradigm was found to be clinically effective in patients with pathological fractures; there was no significant spinal canal compromise. In this technique two minimally invasive surgical procedures are combined to avoid the morbidity associated with open surgery while providing both immediate fracture fixation and administering a single-fraction tumoricidal radiation dose. PMID- 16266072 TI - The therapeutic window for spinal cord decompression in a rat spinal cord injury model. AB - OBJECT: There are no clinically based guidelines to direct the spine surgeon as to the proper timing to undertake decompression after spinal cord injury (SCI) in patients with concomitant stenosis-induced cord compression. The following three factors affect the prognosis: (1) severity of SCI; (2) degree of extrinsic spinal cord compression; and (3) duration of spinal cord compression. METHODS: To elucidate further the relationship between varying degrees of spinal stenosis and a mild contusion-induced SCI (6.25 g-cm), a rat SCI/stenosis model was developed in which 1.13- and 1.24-mm-thick spacers were placed at T-10 to create 38 and 43% spinal stenosis, respectively. Spinal cord damage was observed after the stenosis SCI that was directly proportional to the duration of spinal cord compression. The therapeutic window prior to decompression was 6 and 12 hours in the 43 and 38% stenosis-SCI lesions, respectively, to maintain locomotor activity. A significant difference in total lesion volume was observed between the 2-hour and the delayed time(s) to decompression (38% stenosis-SCI, 12 and 24 hours, p < 0.05; 43% stenosis-SCI, 24 hours, p < 0.05) indicating a more favorable neurological outcome when earlier decompression is undertaken. This finding was further supported by the animal's ability to support weight when decompression was performed by 6 or 12 hours compared with 24 hours after SCI. CONCLUSIONS: Analysis of the findings in this study suggests that early decompression in the rat improves locomotor function. Prolongation of the time to decompression may result in irreversible damage that prevents locomotor recovery. PMID- 16266073 TI - Olfactory glia transplantation into cervical spinal cord contusion injuries. AB - OBJECT: The results of olfactory ensheathing cell (OEC) transplantation have raised great expectations as a potential treatment for spinal cord injury (SCI). Its capacity to promote functional neural repair, however, remains unclear. The authors studied axonal growth and locomotor recovery after C-7 contusion injury and OEC transplantation in adult rats. METHODS: Twenty-four male Wistar rats underwent a mild C-7 contusion injury that completely disrupted the dorsal corticospinal tract (DCST). In 14 rats OECs were transplanted into the lesion, and 10 were used as controls. At 3 months postcontusion, the kinematics of locomotion were assessed, and the CST was traced by injecting dextran tetramethylrhodamine bilaterally into the cerebral cortex. The animals were killed 2 weeks after tracer injection, and their spinal cords were studied immunohistochemically. Although the survival of transplanted cells varied, they were present in all cases. The authors observed neither OEC migration nor DCST axon regeneration in any of the cell transplant-treated rats. Corticospinal axons ended in retraction bulbs at the proximal edge of the lesion or, exceptionally, a few micrometers inside the transplant. The results of neurofilament immunohistochemical analysis provided evidence of neurites from systems other than the DCST growing into the transplant, but in some cases these neurites formed loops of pathological appearance. Contusion injury of C-7 caused chronic locomotor deficits that did not improve after OEC transplants. CONCLUSIONS: The findings in this study indicate that OEC transplants alone are not sufficient for neural repair and functional recovery after SCI. In addition, OECs can induce abnormal axonal growth, making further studies necessary before considering their clinical use. PMID- 16266074 TI - Use of dual transarticular screws to fixate simultaneous occipitoatlantal and atlantoaxial dislocations. AB - Occipitoatlantal dislocation and atlantoaxial vertical distraction are caused by similar mechanisms, and few individuals survive these injuries. It is hypothesized that the injurious vertical force manifests as a traumatic lesion at different levels of the same ligamentous complex. The authors report the cases of two patients who presented with this combined lesion, describe surgical alternatives for stabilization, and introduce a new technique that combines the use of transarticular screws in a "dual" construct, without involving the unaffected spine. PMID- 16266075 TI - Advanced Chiari malformation type I with marked spinal cord atrophy. Case report. AB - The authors discuss the results obtained in patients who underwent foramen magnum decompression for longstanding advanced Chiari I malformation in which marked spinal cord atrophy was present. This 50-year-old woman presented with progressive quadriparesis and sensory disorders. Magnetic resonance imaging revealed the descent of cerebellar tonsils and medulla associated with remarkable C1-L2 spinal cord atrophy. After a C-1 laminectomy-based foramen magnum decompression, arachnoid dissection and duraplasty were undertaken. These procedures resulted in remarkable neurological improvement, even after 40 years of clinical progression. Spinal cord atrophy may be caused by chronic pressure of entrapped cerebrospinal fluid in the spinal canal. PMID- 16266076 TI - A unique two-level noncontiguous ligamentous flexion-distraction injury in the thoracolumbar spine. AB - The authors report the case of a patient who sustained traumatic two-level noncontiguous ligamentous flexion-distraction injuries in the thoracolumbar spine. To the best of their knowledge, this is the first reported case of this combined injury pattern. PMID- 16266077 TI - Bone graft extenders. PMID- 16266078 TI - [Comparative study of daily maximum exposure to temperatures during hot summer days in 3 Japanese cities]. AB - OBJECTIVES: Health risk assessment and developing measures to deal with global warming (including increased heat waves) have become urgent global issues. In the present study, we measured the personally exposed temperature (Tp) during summer among residents in major Japanese cities to investigate the relation to daily maximum ambient temperature (Tmax), which is generally been used as an index of temperature exposure in epidemiological studies. METHODS: Personal exposures to temperature (Tp) were measured for a week with portable monitors (HOBO H8 Loggers, Onset Computer Corporation) for 194 subjects (101 males and 93 females, aged 21-82 years) in 3 cities, i.e., Sapporo, Tokyo and Naha (Okinawa), from July to September, 2003 (73 days). RESULTS AND DISCUSSION: Even on days with a Tmax of 30-35 degrees C, associated with significantly increased risk of mortality in Tokyo, neither average Tp for 7-19 o'clock nor the value for 13-15 o'clock appeared to rise beyond 30 degrees C in Tokyo and 31 degrees C in Naha. It was, thus, apparent that Tp's are generally controlled to not exceed these values at least during daytime, suggesting that they could be regarded as a threshold for heat stress tolerance. On the other hand, although average Tp's for night time (0 7 o'clock) were also found to be asymptomatic at 29 degrees C in Tokyo and 30 degrees C in Naha, they were generally too high to be free from heat stress including sleep disturbance in both cities as indicated in our questionnaire study. For both cities, in days with the Tmax above 30 degrees C, the average Tmin was 26 degrees C, while average and minimum Tp's during the night time were 28 degrees C and 27 degrees C, respectively. The correlation coefficients with Tmax were generally low; 0.35 for average Tp during daytime, 0.42 for Tmin and 0.27 and 0.19 for average and minimum night time Tp, respectively. PMID- 16266080 TI - [Stage progression of dental patients following brief interventions for smoking cessation]. AB - PURPOSE: Potential effects of brief intervention for smoking cessation were evaluated by examination of stage progression with respect to quitting the habit in dental patients. METHODS: Stage progression was retrospectively evaluated in 25 patients undergoing brief interventions since April 2001 at a university dental hospital. Stage of cessation was requested prior to and following interventions (June to December 2003) according to the modification method of Prochaska's model. Brief interventions were conducted by indication of effects of smoking in the mouth and on dental treatment at each visit. Cessation techniques were explained in instances where subjects displayed an interest in smoking cessation. RESULTS: The intervals between dental visits varied (1-6 months). Prior to intervention, numbers of patients in the pre-contemplation, contemplation and preparation stages were 15, 5 and 5, respectively; this changed to 6, 2 and 1, respectively, following intervention, with 16 participants attempting smoking cessation, and 9 reporting continued abstinence. Stage progression was noted in 18 subjects. In the remaining 7 patients, 6 in the pre contemplation and 1 in the contemplation stage, no change was registered. More than half of the patients (11/20) who had not prepared for cessation prior to intervention and all patients (5/5) in the preparation period reported smoking cessation following the brief interventions. CONCLUSION: Brief interventions in dental practice can induce smoking cessation in patients. PMID- 16266079 TI - [Domestic violence against women during pregnancy]. AB - OBJECTIVES: To determine the prevalence of domestic violence (DV) against women during pregnancy and to identify risk factors for DV and effects on women's mental health. METHODS: Pregnant women from an OB-GYN outpatient clinic at a hospital in an urban area were recruited consecutively from February to May 2003. Women who agreed to participate in the research answered three self-administered questionnaires: the GHQ30, the Rosenberg Self Esteem, and one for demographic characteristics at 14 gestational weeks. In order to determine DV prevalence rate during pregnancy, the Japanese version of the Index of Spouse Abuse (ISA) for measuring severity of DV was provided to those women over 35 gestational weeks. RESULTS: Two hundred seventy nine women answered all questionnaires. 15 of the 279 respondents (5.4%) were DV positive during pregnancy based on the Japanese version of the ISA. Nine had experienced physical violence, and twelve had non physical violence Compared with women who had not experienced DV during pregnancy, DV positive pregnant women were more likely to be multipara (OR = 3.9) and to have experienced physical violence in the past from a different partner (OR = 9.1). Moreover, general illness (OR = 3.8), sleep disturbance (OR = 5.8), anxiety (OR = 6.3), depression (OR = 11.5) and low self-esteem (P = 0.02) were identified as effects of DV on women's mental health. CONCLUSION: Some 5.4% of women in Japan, approximately 1 in every 20, may experience DV during pregnancy. This is associated with parity and a past history of DV as demographic characteristics, and has an adverse impact on mental health, especially depression. Development of a support system for screening, intervention and referral for DV sufferers is urgently needed. PMID- 16266082 TI - [Questionnaire study on the use of emergency ambulances in Yokohama, Japan]. PMID- 16266081 TI - [Characteristics of elderly leaders volunteering to participate in a fall prevention programme]. AB - OBJECTIVES: This study was conducted to assess characteristics of elderly leaders volunteering to participate in a fall prevention programme. METHODS: We surveyed 1,503 individuals (75 elderly leaders volunteering to participate in a fall prevention programme and 1,428 non-leader elderly) among the elderly population living in a rural community, Miyagi Prefecture. Subjects were aged 70-84 years. The questionnaire covered socio-demographic factors, as well as physical, psychology and social variables. To analyze the characteristics of the elderly leaders volunteering to participate in this programme, the relationships of socio demographic, physical, psychology and social factors to whether the elderly were leaders in the programme were analyzed using logistic regression. RESULT: As a result of multiple logistic regression analysis, the characteristics of elderly leaders volunteering to participate in the fall prevention programme were as follows; 1) being male (OR = 0.25, 95%CI 0.14-0.44); 2) young age (OR=0.43, 95%CI 0.25-0.73); 3) having a high intellectual activity (OR = 2.72, 95%CI 1.65-4.48); 4) being well satisfied with their health (OR = 1.45, 95%CI 1.02-2.07), and 5) having a high IKIGAI (OR = 1.06, 95%CI 1.01-1.13). CONCLUSION: Only elderly individuals capable of high-level intellectual activities can fill the roles of elderly volunteer group leaders discussed in this study. PMID- 16266083 TI - [Establishment of Healthy Japan 21 Regional plans according to local community type]. AB - OBJECTIVE: To examine the establishment of Health Japan 21 regional plans according to the local community type. METHODS: We sent all the Japanese local governments a questionnaire regarding the establishment of health Japan 21 regional plans with a request to provide a hardcopy of established regional plans. RESULTS: Of the respondent 1954 local communities, the proportions regarding establishment of plans (including "Completed" and "In the course of") according to the community type were: major, middle sized cities and Tokyo metropolitan wards, 100%; other cities, 64.9%; towns, 40.7%; and villages, 38.8%. Of the 500 local communities which answered "Completed establishment", the proportions with release onto the internet (including "Completed" and "In the course of") were: major cities, 100%; middle sized cities, 67.7%; Tokyo metropolitan wards, 85.7%; other cities, 38.8%; towns, 13.5%; and villages, 14.3%. We examined whether each of the 462 collected regional plans included each of the national Health Japan 21's target themes and items, and found a tendency for smaller community' regional plans to be less likely to give comprehensive coverage. CONCLUSION: More support for smaller local governments is necessary to achieve establishment and effective implementation of Health Japan 21 regional plans. PMID- 16266084 TI - [Questionnaire survey of countermeasures against severe acute respiratory syndrome (SARS) in the prefectural governments of Japan]. PMID- 16266085 TI - Deployment of a maintenance of certification program in medical physics serves the interest of the profession and the public. PMID- 16266087 TI - Brachytherapy source characterization for improved dose calculations using primary and scatter dose separation. AB - In brachytherapy, tissue heterogeneities, source shielding, and finite patient/phantom extensions affect both the primary and scatter dose distributions. The primary dose is, due to the short range of secondary electrons, dependent only on the distribution of material located on the ray line between the source and dose deposition site. The scatter dose depends on both the direct irradiation pattern and the distribution of material in a large volume surrounding the point of interest, i.e., a much larger volume must be included in calculations to integrate many small dose contributions. It is therefore of interest to consider different methods for the primary and the scatter dose calculation to improve calculation accuracy with limited computer resources. The algorithms in present clinical use ignore these effects causing systematic dose errors in brachytherapy treatment planning. In this work we review a primary and scatter dose separation formalism (PSS) for brachytherapy source characterization to support separate calculation of the primary and scatter dose contributions. We show how the resulting source characterization data can be used to drive more accurate dose calculations using collapsed cone superposition for scatter dose calculations. Two types of source characterization data paths are used: a direct Monte Carlo simulation in water phantoms with subsequent parameterization of the results, and an alternative data path built on processing of AAPM TG43 formatted data to provide similar parameter sets. The latter path is motivated of the large amounts of data already existing in the TG43 format. We demonstrate the PSS methods using both data paths for a clinical 192Ir source. Results are shown for two geometries: a finite but homogeneous water phantom, and a half-slab consisting of water and air. The dose distributions are compared to results from full Monte Carlo simulations and we show significant improvement in scatter dose calculations when the collapsed-cone kernel-superposition algorithm is used compared to traditional table based calculations. The PSS source characterization method uses exponential fit functions derived from one-dimensional transport theory to describe both the primary and scatter dose contributions. We present data for the PSS characterization method to different 192Ir, 137Cs, and 60Cs brachytherapy sources. We also show how TG43 formatted data can be derived from our data to serve traditional treatment planning systems, as to enable for a gradual transfer to algorithms that provides improved modeling of heterogeneities in brachytherapy treatment planning. PMID- 16266088 TI - A phantom evaluation of a stereo-vision surface imaging system for radiotherapy patient setup. AB - External beam irradiation requires precise positioning of the target relative to the treatment planning coordinate system. A three-dimensional (3D) surface imaging system for patient positioning has recently been installed in one of our linear accelerator (linac) rooms. The device utilizes close-range photogrammetry to generate a 3D model of the patient's surface. This geometric model can be made to look like a digital camera image if wrapped with a gray-level image (texture mapping) that shows surface coloration. The system is calibrated to the linac coordinate system and has been designed as a patient setup device. To reproduce patient position in fractionated radiotherapy, the daily patient surface model is registered to a previously recorded reference surface. Using surface registration, the system calculates the rigid-body transformation that minimizes the distance between the treatment and the reference surface models in a region of-interest (ROI). This transformation is expressed as a set of new couch coordinates at which the patient position best matches with the reference data. If respiratory motion is a concern, the surface can be obtained with a gated acquisition at a specified phase of the respiratory cycle. To analyze the accuracy of the system, we performed several experiments with phantoms to assess stability, alignment accuracy, precision of the gating function, and surface topology. The reproducibility of surface measurements was tested for periods up to 57 h. Each recorded frame was registered to the reference surface to calculate the required couch adjustment. The system stability over this time period was better than 0.5 mm. To measure the accuracy of the system to detect and quantify patient shift relative to a reference image, we compared the shift detected by the surface imaging system with known couch transitions in a phantom study. The maximum standard deviation was 0.75 mm for the three translational degrees of freedom, and less than 0.1 degrees for each rotation. Surface model precision was tested against computed tomography (CT)-derived surface topology. The root-mean square rms of the distance between the surfaces was 0.65 mm, excluding regions where beam hardening caused artifacts in the CT data. Measurements were made to test the gated acquisition mode. The time-dependent amplitude was measured with the surface imaging system and an established respiratory gating system based on infrared (IR)-marker detection. The measured motion trajectories from both systems were compared to the known trajectory of the stage. The standard deviations of the amplitude differences to the motor trajectory were 0.04 and 0.15 mm for the IR-marker system and the 3D surface imaging system, respectively. A limitation of the surface-imaging device is the frame rate of 6.5 Hz, because rapid changes of the motion trajectory cannot be detected. In conclusion, the system is accurate and sufficiently stable to be used in the clinic. The errors computed when comparing the surface model with CT geometry were submillimeter, and deviations in the alignment and gating-signal tests were of the same magnitude. PMID- 16266089 TI - Automatic exposure control for a slot scanning full field digital mammography system. AB - Automatic exposure control (AEC) is an important feature in mammography. It enables consistently optimal image exposure despite variations in tissue density and thickness, and user skill level. Full field digital mammography systems cannot employ conventional AEC methods because digital receptors fully absorb the x-ray beam. In this paper we describe an AEC procedure for slot scanning mammography. With slot scanning detectors, our approach uses a fast low resolution and low-exposure prescan to acquire an image of the breast. Tube potential depends on breast thickness, and the prescan histogram provides the necessary information to calculate the required tube current. We validate our approach with simulated prescan images and phantom measurements. We achieve accurate exposure tracking with thickness and density, and expect this method of AEC to reduce retakes and improve workflow. PMID- 16266090 TI - 4D rotational x-ray imaging of wrist joint dynamic motion. AB - Current methods for imaging joint motion are limited to either two-dimensional (2D) video fluoroscopy, or to animated motions from a series of static three dimensional (3D) images. 3D movement patterns can be detected from biplane fluoroscopy images matched with computed tomography images. This involves several x-ray modalities and sophisticated 2D to 3D matching for the complex wrist joint. We present a method for the acquisition of dynamic 3D images of a moving joint. In our method a 3D-rotational x-ray (3D-RX) system is used to image a cyclically moving joint. The cyclic motion is synchronized to the x-ray acquisition to yield multiple sets of projection images, which are reconstructed to a series of time resolved 3D images, i.e., four-dimensional rotational x ray (4D-RX). To investigate the obtained image quality parameters the full width at half maximum (FWHM) of the point spread function (PSF) via the edge spread function and the contrast to noise ratio between air and phantom were determined on reconstructions of a bullet and rod phantom, using 4D-RX as well as stationary 3D RX images. The CNR in volume reconstructions based on 251 projection images in the static situation and on 41 and 34 projection images of a moving phantom were 6.9, 3.0, and 2.9, respectively. The average FWHM of the PSF of these same images was, respectively, 1.1, 1.7, and 2.2 mm orthogonal to the motion and parallel to direction of motion 0.6, 0.7, and 1.0 mm. The main deterioration of 4D-RX images compared to 3D-RX images is due to the low number of projection images used and not to the motion of the object. Using 41 projection images seems the best setting for the current system. Experiments on a postmortem wrist show the feasibility of the method for imaging 3D dynamic joint motion. We expect that 4D RX will pave the way to improved assessment of joint disorders by detection of 3D dynamic motion patterns in joints. PMID- 16266091 TI - Statistical process control for radiotherapy quality assurance. AB - Every quality assurance process uncovers random and systematic errors. These errors typically consist of many small random errors and a very few number of large errors that dominate the result. Quality assurance practices in radiotherapy do not adequately differentiate between these two sources of error. The ability to separate these types of errors would allow the dominant source(s) of error to be efficiently detected and addressed. In this work, statistical process control is applied to quality assurance in radiotherapy for the purpose of setting action thresholds that differentiate between random and systematic errors. The theoretical development and implementation of process behavior charts are described. We report on a pilot project is which these techniques are applied to daily output and flatness/symmetry quality assurance for a 10 MV photon beam in our department. This clinical case was followed over 52 days. As part of our investigation, we found that action thresholds set using process behavior charts were able to identify systematic changes in our daily quality assurance process. This is in contrast to action thresholds set using the standard deviation, which did not identify the same systematic changes in the process. The process behavior thresholds calculated from a subset of the data detected a 2% change in the process whereas with a standard deviation calculation, no change was detected. Medical physicists must make decisions on quality assurance data as it is acquired. Process behavior charts help decide when to take action and when to acquire more data before making a change in the process. PMID- 16266092 TI - Refraction-enhanced tomography of mouse and rabbit lungs. AB - In order to evaluate the effectiveness of edge enhancement by refraction in computed tomography, images of a cross section of a euthanized mouse thorax were recorded at low (20 keV) and high (72 keV) x-ray energies at a spatial resolution of about 40 microm. Compared with the images obtained with the detector at 30 cm from an object, when the object was located at 113 cm from the detector, the contrast between tissues and air was improved at both energies. The improvement was more pronounced at 72 keV where the absorption contrast was weaker. This effect was due to refraction at the surfaces of alveolar membranes and small airways which creates areas with apparently high and low linear attenuation coefficients within tissues. The edge enhancement by refraction was also effective in images of a euthanized rabbit thorax at x-ray energies of 40 and 70 keV at a spatial resolution of about 0.15 mm. These results raise the possibility that the refraction contrast may be utilized to obtain a high-resolution tomographic image of human lung and bone with low dose. PMID- 16266093 TI - Optimizing the acquisition time profile for a planar integral measurement system with a spinning slat collimator. AB - This article considers a hypothetical imaging device with a spinning slat collimator that measures parallel-planar-integral data from an object. This device rotates around the object 180 degrees and stops at N positions uniformly distributed over this 180 degrees. At each stop, the device spins on its own axis 180 degrees and acquires measurements at M positions uniformly distributed over this 180 degrees. For a fixed total imaging time, an optimal distribution of the scanning time among the data measurement locations is searched by a nonlinear programming method: Nelder-Mead's simplex method. The optimal dwell time is approximately proportional to the weighting factor in the backprojector of the reconstruction algorithm. By using an optimal dwell-time profile, the reconstruction signal-to-noise ratio has a gain of 23%-24% for the filtered backprojection algorithm and a gain of 10%-18% for the iterative algorithms, compared with the situation when a constant dwell-time profile is used. PMID- 16266094 TI - Attenuation correction for small animal SPECT imaging using x-ray CT data. AB - Photon attenuation in small animal nuclear medicine scans can be significant when using isotopes that emit lower energy photons such as iodine-125. We have developed a method to use microCT data to perform attenuation corrected small animal single-photon emission computed tomography (SPECT). A microCT calibration phantom was first imaged, and the resulting calibration curve was used to convert microCT image values to linear attenuation coefficient values that were then used in an iterative SPECT reconstruction algorithm. This method was applied to reconstruct a SPECT image of a uniform phantom filled with 125I-NaI. Without attenuation correction, the image suffered a 30% decrease in intensity in the center of the image, which was removed with the addition of attenuation correction. This reduced the relative standard deviation in the region of interest from 10% to 6%. PMID- 16266095 TI - Experimental verification of a portal dose prediction model. AB - Electronic portal imaging devices (EPIDs) can be used to measure a two dimensional (2D) dose distribution behind a patient, thus allowing dosimetric treatment verification. For this purpose we experimentally assessed the accuracy of a 2D portal dose prediction model based on pencil beam scatter kernels. A straightforward derivation of these pencil beam scatter kernels for portal dose prediction models is presented based on phantom measurements. The model is able to predict the 2D portal dose image (PDI) behind a patient, based on a PDI without the patient in the beam in combination with the radiological thickness of the patient, which requires in addition a PDI with the patient in the beam. To assess the accuracy of portal dose and radiological thickness values obtained with our model, various types of homogeneous as well as inhomogeneous phantoms were irradiated with a 6 MV photon beam. With our model we are able to predict a PDI with an accuracy better than 2% (mean difference) if the radiological thickness of the object in the beam is symmetrically situated around the isocenter. For other situations deviations up to 3% are observed for a homogeneous phantom with a radiological thickness of 17 cm and a 9 cm shift of the midplane-to-detector distance. The model can extract the radiological thickness within 7 mm (maximum difference) of the actual radiological thickness if the object is symmetrically distributed around the isocenter plane. This difference in radiological thickness is related to a primary portal dose difference of 3%. It can be concluded that our model can be used as an easy and accurate tool for the 2D verification of patient treatments by comparing predicted and measured PDIs. The model is also able to extract the primary portal dose with a high accuracy, which can be used as the input for a 3D dose reconstruction method based on back-projection. PMID- 16266096 TI - A technique for on-board CT reconstruction using both kilovoltage and megavoltage beam projections for 3D treatment verification. AB - The technologies with kilovoltage (kV) and megavoltage (MV) imaging in the treatment room are now available for image-guided radiation therapy to improve patient setup and target localization accuracy. However, development of strategies to efficiently and effectively implement these technologies for patient treatment remains challenging. This study proposed an aggregated technique for on-board CT reconstruction using combination of kV and MV beam projections to improve the data acquisition efficiency and image quality. These projections were acquired in the treatment room at the patient treatment position with a new kV imaging device installed on the accelerator gantry, orthogonal to the existing MV portal imaging device. The projection images for a head phantom and a contrast phantom were acquired using both the On-Board Imager kV imaging device and the MV portal imager mounted orthogonally on the gantry of a Varian Clinac 21EX linear accelerator. MV projections were converted into kV information prior to the aggregated CT reconstruction. The multilevel scheme algebraic reconstruction technique was used to reconstruct CT images involving either full, truncated, or a combination of both full and truncated projections. An adaptive reconstruction method was also applied, based on the limited numbers of kV projections and truncated MV projections, to enhance the anatomical information around the treatment volume and to minimize the radiation dose. The effects of the total number of projections, the combination of kV and MV projections, and the beam truncation of MV projections on the details of reconstructed kV/MV CT images were also investigated. PMID- 16266098 TI - Spatiotemporal processing of gated cardiac SPECT images using deformable mesh modeling. AB - In this paper we present a spatiotemporal processing approach, based on deformable mesh modeling, for noise reduction in gated cardiac single-photon emission computed tomography images. Because of the partial volume effect (PVE), clinical cardiac-gated perfusion images exhibit a phenomenon known as brightening the myocardium appears to become brighter as the heart wall thickens. Although brightening is an artifact, it serves as an important diagnostic feature for assessment of wall thickening in clinical practice. Our proposed processing algorithm aims to preserve this important diagnostic feature while reducing the noise level in the images. The proposed algorithm is based on the use of a deformable mesh for modeling the cardiac motion in a gated cardiac sequence, based on which the images are processed by smoothing along space-time trajectories of object points while taking into account the PVE. Our experiments demonstrate that the proposed algorithm can yield significantly more-accurate results than several existing methods. PMID- 16266097 TI - Computer-aided detection of breast masses on full field digital mammograms. AB - We are developing a computer-aided detection (CAD) system for breast masses on full field digital mammographic (FFDM) images. To develop a CAD system that is independent of the FFDM manufacturer's proprietary preprocessing methods, we used the raw FFDM image as input and developed a multiresolution preprocessing scheme for image enhancement. A two-stage prescreening method that combines gradient field analysis with gray level information was developed to identify mass candidates on the processed images. The suspicious structure in each identified region was extracted by clustering-based region growing. Morphological and spatial gray-level dependence texture features were extracted for each suspicious object. Stepwise linear discriminant analysis (LDA) with simplex optimization was used to select the most useful features. Finally, rule-based and LDA classifiers were designed to differentiate masses from normal tissues. Two data sets were collected: a mass data set containing 110 cases of two-view mammograms with a total of 220 images, and a no-mass data set containing 90 cases of two-view mammograms with a total of 180 images. All cases were acquired with a GE Senographe 2000D FFDM system. The true locations of the masses were identified by an experienced radiologist. Free-response receiver operating characteristic analysis was used to evaluate the performance of the CAD system. It was found that our CAD system achieved a case-based sensitivity of 70%, 80%, and 90% at 0.72, 1.08, and 1.82 false positive (FP) marks/image on the mass data set. The FP rates on the no-mass data set were 0.85, 1.31, and 2.14 FP marks/image, respectively, at the corresponding sensitivities. This study demonstrated the usefulness of our CAD techniques for automated detection of masses on FFDM images. PMID- 16266099 TI - The application of the sinusoidal model to lung cancer patient respiratory motion. AB - Accurate modeling of the respiratory cycle is important to account for the effect of organ motion on dose calculation for lung cancer patients. The aim of this study is to evaluate the accuracy of a respiratory model for lung cancer patients. Lujan et al. [Med. Phys. 26(5), 715-720 (1999)] proposed a model, which became widely used, to describe organ motion due to respiration. This model assumes that the parameters do not vary between and within breathing cycles. In this study, first, the correlation of respiratory motion traces with the model f(t) as a function of the parameter n (n = 1, 2, 3) was undertaken for each breathing cycle from 331 four-minute respiratory traces acquired from 24 lung cancer patients using three breathing types: free breathing, audio instruction, and audio-visual biofeedback. Because cos2 and cos4 had similar correlation coefficients, and cos2 and cos1 have a trigonometric relationship, for simplicity, the cos1 value was consequently used for further analysis in which the variations in mean position (z0), amplitude of motion (b) and period (tau) with and without biofeedback or instructions were investigated. For all breathing types, the parameter values, mean position (z0), amplitude of motion (b), and period (tau) exhibited significant cycle-to-cycle variations. Audio-visual biofeedback showed the least variations for all three parameters (z0, b, and tau). It was found that mean position (z0) could be approximated with a normal distribution, and the amplitude of motion (b) and period (tau) could be approximated with log normal distributions. The overall probability density function (pdf) of f(t) for each of the three breathing types was fitted with three models: normal, bimodal, and the pdf of a simple harmonic oscillator. It was found that the normal and the bimodal models represented the overall respiratory motion pdfs with correlation values from 0.95 to 0.99, whereas the range of the simple harmonic oscillator pdf correlation values was 0.71 to 0.81. This study demonstrates that the pdfs of mean position (z0), amplitude of motion (b), and period (tau) can be used for sampling to obtain more realistic respiratory traces. The overall standard deviations of respiratory motion were 0.48, 0.57, and 0.55 cm for free breathing, audio instruction, and audio-visual biofeedback, respectively. PMID- 16266100 TI - Effects of point configuration on the accuracy in 3D reconstruction from biplane images. AB - Two or more angiograms are being used frequently in medical imaging to reconstruct locations in three-dimensional (3D) space, e.g., for reconstruction of 3D vascular trees, implanted electrodes, or patient positioning. A number of techniques have been proposed for this task. In this simulation study, we investigate the effect of the shape of the configuration of the points in 3D (the "cloud" of points) on reconstruction errors for one of these techniques developed in our laboratory. Five types of configurations (a ball, an elongated ellipsoid (cigar), flattened ball (pancake), flattened cigar, and a flattened ball with a single distant point) are used in the evaluations. For each shape, 100 random configurations were generated, with point coordinates chosen from Gaussian distributions having a covariance matrix corresponding to the desired shape. The 3D data were projected into the image planes using a known imaging geometry. Gaussian distributed errors were introduced in the x and y coordinates of these projected points. Gaussian distributed errors were also introduced into the gantry information used to calculate the initial imaging geometry. The imaging geometries and 3D positions were iteratively refined using the enhanced-Metz Fencil technique. The image data were also used to evaluate the feasible R-t solution volume. The 3D errors between the calculated and true positions were determined. The effects of the shape of the configuration, the number of points, the initial geometry error, and the input image error were evaluated. The results for the number of points, initial geometry error, and image error are in agreement with previously reported results, i.e., increasing the number of points and reducing initial geometry and/or image error, improves the accuracy of the reconstructed data. The shape of the 3D configuration of points also affects the error of reconstructed 3D configuration; specifically, errors decrease as the "volume" of the 3D configuration increases, as would be intuitively expected, and shapes with larger spread, such as spherical shapes, yield more accurate reconstructions. These results are in agreement with an analysis of the solution volume of feasible geometries and could be used to guide selection of points for reconstruction of 3D configurations from two views. PMID- 16266101 TI - Progressive attenuation fields: fast 2D-3D image registration without precomputation. AB - Computation of digitally reconstructed radiograph (DRR) images is the rate limiting step in most current intensity-based algorithms for the registration of three-dimensional (3D) images to two-dimensional (2D) projection images. This paper introduces and evaluates the progressive attenuation field (PAF), which is a new method to speed up DRR computation. A PAF is closely related to an attenuation field (AF). A major difference is that a PAF is constructed on the fly as the registration proceeds; it does not require any precomputation time, nor does it make any prior assumptions of the patient pose or limit the permissible range of patient motion. A PAF effectively acts as a cache memory for projection values once they are computed, rather than as a lookup table for precomputed projections like standard AFs. We use a cylindrical attenuation field parametrization, which is better suited for many medical applications of 2D-3D registration than the usual two-plane parametrization. The computed attenuation values are stored in a hash table for time-efficient storage and access. Using clinical gold-standard spine image data sets from five patients, we demonstrate consistent speedups of intensity-based 2D-3D image registration using PAF DRRs by a factor of 10 over conventional ray casting DRRs with no decrease of registration accuracy or robustness. PMID- 16266102 TI - Measurement of CT radiation profile width using CR imaging plates. AB - This paper describes the procedure for using a Fuji computed radiography (CR) imaging plate (IP) for the measurement of computed tomography (CT) radiation profiles. Two sources of saturation in the data from the IP, signal and quantization, were characterized to establish appropriate exposure and processing conditions for accurate measurements. The IP generated similar profiles compared to those obtained from digitized ready-pack films, except at the profile edges, where the exposure level is low. However, when IP pixel values are converted to exposure, CR and digitized film profiles are in agreement. The full width at half maximum (FWHM) of the CT radiation profile was determined from the relationship between pixel value and exposure and compared to FWHM of the digitized optical density profile from film. To estimate the effect of scattering by the cassette material, radiation profiles were acquired from IPs enclosed in a cassette or in a paper envelope. The presence of the cassette made no difference in the value determined for FWHM. With proper exposure and processing conditions, the FWHM of 5, 10, and 15 mm collimated beams were measured using IPs to be 7.1, 11.9, and 17.0 mm and using film to be 7.2, 12.2, and 16.8 mm, respectively. Our results suggest that, under appropriate conditions, the estimation of the width of the CT radiation profile using Fuji CR is comparable to the measurement from film density described in American Association of Physicists in Medicine (AAPM) Report No. 39. Although our experiment was conducted using Fuji CR, we anticipate that CR plates from other vendors could be successfully used to measure CT beam profiles because of similar empirical relationships between pixel value and exposure. PMID- 16266103 TI - Prospective respiratory-gated micro-CT of free breathing rodents. AB - Microcomputed tomography (Micro-CT) has the potential to noninvasively image the structure of organs in rodent models with high spatial resolution and relatively short image acquisition times. However, motion artifacts associated with the normal respiratory motion of the animal may arise when imaging the abdomen or thorax. To reduce these artifacts and the accompanying loss of spatial resolution, we propose a prospective respiratory gating technique for use with anaesthetized, free-breathing rodents. A custom-made bed with an embedded pressure chamber was connected to a pressure transducer. Anaesthetized animals were placed in the prone position on the bed with their abdomens located over the chamber. During inspiration, the motion of the diaphragm caused an increase in the chamber pressure, which was converted into a voltage signal by the transducer. An output voltage was used to trigger image acquisition at any desired time point in the respiratory cycle. Digital radiographic images were acquired of anaesthetized, free-breathing rats with a digital radiographic system to correlate the respiratory wave form with respiration-induced organ motion. The respiratory wave form was monitored and recorded simultaneously with the x-ray radiation pulses, and an imaging window was defined, beginning at end expiration. Phantom experiments were performed to verify that the respiratory gating apparatus was triggering the micro-CT system. Attached to the distensible phantom were 100 microm diameter copper wires and the measured full width at half maximum was used to assess differences in image quality between respiratory-gated and ungated imaging protocols. This experiment allowed us to quantify the improvement in the spatial resolution, and the reduction of motion artifacts caused by moving structures, in the images resulting from respiratory-gated image acquisitions. The measured wire diameters were 0.135 mm for the stationary phantom image, 0.137 mm for the image gated at end deflation, 0.213 mm for the image gated at peak inflation, and 0.406 mm for the ungated image. Micro-CT images of anaesthetized, free-breathing rats were acquired with a General Electric Healthcare eXplore RS in vivo micro-CT system. Images of the thorax were acquired using the respiratory cycle-based trigger for the respiratory-gated mode. Respiratory gated-images were acquired at inspiration and end expiration, during a period of minimal respiration-induced organ motion. Gated images were acquired with a nominal isotropic voxel spacing of 44 microm in 20-25 min (80 kVp, 113 mAs, 300 ms imaging window per projection). The equivalent ungated acquisitions were 11 min in length. We observed improved definition of the diaphragm boundary and increased conspicuity of small structures within the lungs in the gated images, when compared to the ungated acquisitions. In this work, we have characterized the externally monitored respiratory wave form of free-breathing, anaesthetized rats and correlated the respiration-induced organ motion to the respiratory cycle. We have shown that the respiratory pressure wave form is an excellent surrogate for the radiographic organ motion. This information facilitates the definition of an imaging window at any phase of the breathing cycle. This approach for prospectively gated micro-CT can provide high quality images of anaesthetized free-breathing rodents. PMID- 16266104 TI - Characteristics of induced activity from medical linear accelerators. AB - A study of the induced activity in a medical linear accelerator (linac) room was carried out on several linac installations. Higher beam energy, higher dose rate, and larger field size generally result in higher activation levels at a given point of interest, while the use of multileaf collimators (MLC) can also increase the activation level at the isocenter. Both theoretical and experimental studies reveal that the activation level in the morning before any clinical work increases from Monday to Saturday and then decreases during the weekend. This weekly activation picture keeps stable from one week to another during standard clinical operation of the linac. An effective half-life for a given point in the treatment room can be determined from the measured or calculated activity decay curves. The effective half-life for points inside the treatment field is longer than that for points outside of the field in the patient plane, while a larger field and longer irradiation time can also make the effective half-life longer. The activation level reaches its practical saturation value after a 30 min continuous irradiation, corresponding to 12 000 MU at a "dose rate" of 400 MU/min. A "dose" of 300 MU was given 20 times in 15 min intervals to determine the trends in the activation level in a typical clinical mode. As well, a long term (85 h over a long weekend) decay curve was measured to evaluate the long term decay of room activation after a typical day of clinical linac use. A mathematical model for the activation level at the isocenter has been established and shown to be useful in explaining and predicting the induced activity levels for typical clinical and experimental conditions. The activation level for a 22 MeV electron beam was also measured and the result shows it is essentially negligible. PMID- 16266105 TI - Time-frequency analysis of the second cardiac sound in phonocardiogram signals. AB - The paper is concerned with the analysis of the phonocardiogram signals (PCG) in the time-frequency domain. Three techniques are studied and evaluated in PCG signal analysis. These are the short time Fourier transform (STFT), the Wigner distribution function (WD) and the continuous wavelet transforms (CWTs). The analysis is first carried out on the second cardiac sound (S2) in order to show the aptitude of each method in distinguishing the internal components of this sound. The results we obtain show that the STFT cannot detect the two internal components of S2 (A2 and P2, respectively, the aortic and pulmonary components). The WD can provide time-frequency characteristics of S2, but with insufficient diagnostic information: the two components are not accurately detected and appear to be only one component. It is found that the CWT (it can also provide the time frequency characteristic of S2) is capable of detecting its two components, A2 and P2, allowing therefore the measurement of the delay between them. This delay, called the split, is very important in the diagnosis of many pathological cases, as it is emphasized in the results we obtain by applying the CWT on different pathological cases (mitral stenosis, pulmonary stenosis and atrial septal defect). PMID- 16266106 TI - Influence of ion chamber response on in-air profile measurements in megavoltage photon beams. AB - This article presents an investigation of the influence of the ion chamber response, including buildup caps, on the measurement of in-air off-axis ratio (OAR) profiles in megavoltage photon beams using Monte Carlo simulations with the EGSnrc system. Two new techniques for the calculation of OAR profiles are presented. Results of the Monte Carlo simulations are compared to measurements performed in 6, 10 and 25 MV photon beams produced by an Elekta Precise linac and shown to agree within the experimental and simulation uncertainties. Comparisons with calculated in-air kerma profiles demonstrate that using a plastic mini phantom gives more accurate air-kerma measurements than using high-Z material buildup caps and that the variation of chamber response with distance from the central axis must be taken into account. PMID- 16266107 TI - Oblique needle segmentation and tracking for 3D TRUS guided prostate brachytherapy. AB - An algorithm was developed in order to segment and track brachytherapy needles inserted along oblique trajectories. Three-dimensional (3D) transrectal ultrasound (TRUS) images of the rigid rod simulating the needle inserted into the tissue-mimicking agar and chicken breast phantoms were obtained to test the accuracy of the algorithm under ideal conditions. Because the robot possesses high positioning and angulation accuracies, we used the robot as a "gold standard," and compared the results of algorithm segmentation to the values measured by the robot. Our testing results showed that the accuracy of the needle segmentation algorithm depends on the needle insertion distance into the 3D TRUS image and the angulations with respect to the TRUS transducer, e.g., at a 10 degrees insertion anglulation in agar phantoms, the error of the algorithm in determining the needle tip position was less than 1 mm when the insertion distance was greater than 15 mm. Near real-time needle tracking was achieved by scanning a small volume containing the needle. Our tests also showed that, the segmentation time was less than 60 ms, and the scanning time was less than 1.2 s, when the insertion distance into the 3D TRUS image was less than 55 mm. In our needle tracking tests in chicken breast phantoms, the errors in determining the needle orientation were less than 2 degrees in robot yaw and 0.7 degrees in robot pitch orientations, for up to 20 degrees needle insertion angles with the TRUS transducer in the horizontal plane when the needle insertion distance was greater than 15 mm. PMID- 16266108 TI - A novel magnetic method for examination of bowel motility. AB - In contrast to the well-developed methods for morphological diagnosis of the gastrointestinal tract, there is no comparatively satisfying technique for functional disorders. One important example is irritable bowel syndrome (IBS), a disorder that affects a high percentage of all individuals. It can only be diagnosed by excluding organic diseases and by considering symptom criteria. In this case, the examination of the motility of the bowel may be a promising way to differentiate between the two major mechanisms of IBS: increased sensitivity of the intestine and altered gastrointestinal motility. To this aim, a recently developed method for monitoring magnetic markers in the gastrointestinal tract was utilized that works without the use of ionizing radiation. We give a short description of this method, showing a spatial resolution of 3-4 mm and a temporal resolution of 330 ms, and report on examples of the first in vivo experiments. Typical monitoring results are shown for the esophagus, the stomach, and the duodenum. The motility behavior is described for the lower parts of the gut as well. The advantages and drawbacks of this type of magnetic marker monitoring are discussed with special consideration of the noninvasive examination of the motility in different sections of the gut. PMID- 16266109 TI - Absorbed dose to water reference dosimetry using solid phantoms in the context of absorbed-dose protocols. AB - For reasons of phantom material reproducibility, the absorbed dose protocols of the American Association of Physicists in Medicine (AAPM) (TG-51) and the International Atomic Energy Agency (IAEA) (TRS-398) have made the use of liquid water as a phantom material for reference dosimetry mandatory. In this work we provide a formal framework for the measurement of absorbed dose to water using ionization chambers calibrated in terms of absorbed dose to water but irradiated in solid phantoms. Such a framework is useful when there is a desire to put dose measurements using solid phantoms on an absolute basis. Putting solid phantom measurements on an absolute basis has distinct advantages in verification measurements and quality assurance. We introduce a phantom dose conversion factor that converts a measurement made in a solid phantom and analyzed using an absorbed dose calibration protocol into absorbed dose to water under reference conditions. We provide techniques to measure and calculate the dose transfer from solid phantom to water. For an Exradin A12 ionization chamber, we measured and calculated the phantom dose conversion factor for six Solid Water phantoms and for a single Lucite phantom for photon energies between 60Co and 18 MV photons. For Solid Water of certified grade, the difference between measured and calculated factors varied between 0.0% and 0.7% with the average dose conversion factor being low by 0.4% compared with the calculation whereas for Lucite, the agreement was within 0.2% for the one phantom examined. The composition of commercial plastic phantoms and their homogeneity may not always be reproducible and consistent with assumed composition. By comparing measured and calculated phantom conversion factors, our work provides methods to verify the consistency of a given plastic for the purpose of clinical reference dosimetry. PMID- 16266110 TI - Indirect flat-panel detector with avalanche gain: fundamental feasibility investigation for SHARP-AMFPI (scintillator HARP active matrix flat panel imager). AB - An indirect flat-panel imager (FPI) with avalanche gain is being investigated for low-dose x-ray imaging. It is made by optically coupling a structured x-ray scintillator CsI(Tl) to an amorphous selenium (a-Se) avalanche photoconductor called HARP (high-gain avalanche rushing photoconductor). The final electronic image is read out using an active matrix array of thin film transistors (TFT). We call the proposed detector SHARP-AMFPI (scintillator HARP active matrix flat panel imager). The advantage of the SHARP-AMFPI is its programmable gain, which can be turned on during low dose fluoroscopy to overcome electronic noise, and turned off during high dose radiography to avoid pixel saturation. The purpose of this paper is to investigate the important design considerations for SHARP-AMFPI such as avalanche gain, which depends on both the thickness d(Se) and the applied electric field E(Se) of the HARP layer. To determine the optimal design parameter and operational conditions for HARP, we measured the E(Se) dependence of both avalanche gain and optical quantum efficiency of an 8 microm HARP layer. The results were used in a physical model of HARP as well as a linear cascaded model of the FPI to determine the following x-ray imaging properties in both the avalanche and nonavalanche modes as a function of E(Se): (1) total gain (which is the product of avalanche gain and optical quantum efficiency); (2) linearity; (3) dynamic range; (4) gain nonuniformity resulting from thickness nonuniformity; and (5) effects of direct x-ray interaction in HARP. Our results showed that a HARP layer thickness of 8 microm can provide adequate avalanche gain and sufficient dynamic range for x-ray imaging applications to permit quantum limited operation over the range of exposures needed for radiography and fluoroscopy. PMID- 16266111 TI - Evaluation of x-ray scatter properties in a dedicated cone-beam breast CT scanner. AB - The magnitude of scatter contamination on a first-generation prototype breast computed tomography (CT) scanner was evaluated using the scatter-to-primary ratio (SPR) metric. The SPR was measured and characterized over a wide range of parameters relevant to breast CT imaging, including x-ray beam energy, breast diameter, breast composition, isocenter-to-detector distance, collimated slot thickness, and grid ratio. The results demonstrated that in the absence of scatter reduction techniques, the SPR levels for the average breast (e.g., 14 cm diameter 50/50 composition cylindrical phantom) are quite high (approximately 0.5 at the center of the phantom for 80 kVp in true cone-beam CT geometry), and increases as the diameter of the phantom is increased (to approximately 1.0 at the center of a 18 cm diameter 50/50 phantom). The x-ray beam energy and the phantom compositions had only minimal impact on the measured SPR. When an ideal bowtie filter was used, the SPRs at the central axis of the 14 and 18 cm cylindrical phantoms were reduced while the SPRs at the edge of the phantoms were increased. Lastly, collimation in the vertical direction had a significant impact on the SPRs at the central axis of the phantoms. These high SPR levels might lead to cupping artifacts and increased noise in the reconstructed CT images, and this suggests that efficient scatter rejection and/or correction techniques may be required to improve the quality and accuracy of cone beam CT images. PMID- 16266112 TI - Energy modulated electron therapy using a few leaf electron collimator in combination with IMRT and 3D-CRT: Monte Carlo-based planning and dosimetric evaluation. AB - Energy modulated electron therapy (EMET) based on Monte Carlo dose calculation is a promising technique that enhances the treatment planning and delivery of superficially located tumors. This study investigated the application of EMET using a novel few-leaf electron collimator (FLEC) in head and neck and breast sites in comparison with three-dimensional conventional radiation therapy (3D CRT) and intensity modulated radiation therapy (IMRT) techniques. Treatment planning was performed for two parotid cases and one breast case. Four plans were compared for each case: 3D-CRT, IMRT, 3D-CRT in conjunction with EMET (EMET-CRT), and IMRT in conjunction with EMET (EMET-IMRT), all of which were performed and calculated with Monte Carlo techniques. For all patients, dose volume histograms (DVHs) were obtained for all organs of interest and the DVHs were used as a means of comparing the plans. Homogeneity and conformity of dose distributions were calculated, as well as a sparing index that compares the effect of the low isodose lines. In addition, the whole-body dose equivalent (WBDE) was estimated for each plan. Adding EMET delivered with the FLEC to 3D-CRT improves sparing of normal tissues. For the two head and neck cases, the mean dose to the contralateral parotid and brain stem was reduced relative to IMRT by 43% and 84%, and by 57% and 71%, respectively. Improved normal tissue sparing was quantified as an increase in sparing index of 47% and 30% for the head and neck and the breast cases, respectively. Adding EMET to either 3D-CRT or IMRT results in preservation of target conformity and dose homogeneity. When adding EMET to the treatment plan, the WBDE was reduced by between 6% and 19% for 3D-CRT and by between 21% and 33% for IMRT, while WBDE for EMET-CRT was reduced by up to 72% when compared with IMRT. FLEC offers a practical means of delivering modulated electron therapy. Although adding EMET delivered using the FLEC results in perturbation of target conformity when compared to IMRT, it significantly improves normal tissue sparing while offering enhanced target conformity to the 3D-CRT planning. The addition of EMET systematically leads to a reduction in WBDE especially when compared with IMRT. PMID- 16266113 TI - Evaluation of a computed radiography system for megavoltage photon beam dosimetry. AB - Computed radiography (CR) systems have been gaining adoption as digital replacements for film for diagnostic and therapy imaging. As a result, film processors are being removed from service, leaving a void for the medical physicists who use film and processors for two-dimensional mega-voltage beam dosimetry. This is the first report to evaluate the ability of a commercial CR reader and storage phosphor plate system to accurately quantitate absolute dose and dose distributions from a 6 MV photon beam. There are potential advantages and disadvantages of current CR systems compared to film systems. CR systems inherently produce a linear dose-response over several logs of dose. However, the barium in the storage phosphor has a higher atomic number than the silver in film, resulting in significant energy sensitivity. The purpose of this work is to fully characterize the impact of these and other features of this CR system relevant to dosimetry. The tests performed and reported on in this study include uniformity of readout across a uniform field, geometrical accuracy, intra- and interday reproducibility, signal decay with time and with light exposure, dose-to signal calibration, high dose effects, obliquity effects, perpendicular and parallel calibration results, field size and depth of measurement effects and the use of lead filters to minimize them, and intensity modulated radiation therapy quality assurance test results compared to that for film. Practical techniques are provided to optimize the accuracy of the system as a dosimetric replacement for film. PMID- 16266114 TI - Spectral discrimination of Cerenkov radiation in scintillating dosimeters. AB - Radiation therapy accelerators require highly accurate dose deposition and the output must be monitored frequently and regularly. Ionization chambers are the primary tool for this control, but their size, their high voltage needed, and the correction needed for electrons make them unsuitable for use during patient treatment. We have developed a small (1-mm-diam and 1-mm-long active part), flexible, and water-equivalent dosimeter. It is suitable for photon and electron beams without corrections, and performs on line dose measurements. This detector is based on only one scintillating fiber and a CCD camera. A new signal processing is used to remove the effect of Cerenkov radiation background, which only requires a preliminary calibration. Central-axis depth-dose distribution comparisons have been achieved with standard ionization chambers, over a range from 8 to 25 MV photons and from 6 to 21 MeV electrons in order to validate this calibration. Results show a very good agreement, with less than 1% difference between the two detectors. PMID- 16266116 TI - Statistical cerebrovascular segmentation in three-dimensional rotational angiography based on maximum intensity projections. AB - Segmentation of three-dimensional rotational angiography (3D-RA) can provide quantitative 3D morphological information of vasculature. The expectation maximization-(EM-) based segmentation techniques have been widely used in the medical image processing community, because of the implementation simplicity, and computational efficiency of the approach. In a brain 3D-RA, vascular regions usually occupy a very small proportion (around 1%) inside an entire image volume. This severe imbalance between the intensity distributions of vessels and background can lead to inaccurate statistical modeling in the EM-based segmentation methods, and thus adversely affect the segmentation quality for 3D RA. In this paper we present a new method for the extraction of vasculature in 3D RA images. The new method is fully automatic and computationally efficient. As compared with the original 3D-RA volume, there is a larger proportion (around 20%) of vessels in its corresponding maximum intensity projection (MIP) image. The proposed method exploits this property to increase the accuracy of statistical modeling with the EM algorithm. The algorithm takes an iterative approach to compiling the 3D vascular segmentation progressively with the segmentation of MIP images along the three principal axes, and use a winner-takes all strategy to combine the results obtained along individual axes. Experimental results on 12 3D-RA clinical datasets indicate that the segmentations obtained by the new method exhibit a high degree of agreement to the ground truth segmentations and are comparable to those produced by the manual optimal global thresholding method. PMID- 16266115 TI - Feasibility of a fast inverse dose optimization algorithm for IMRT via matrix inversion without negative beamlet intensities. AB - A fast optimization algorithm is very important for inverse planning of intensity modulated radiation therapy (IMRT), and for adaptive radiotherapy of the future. Conventional numerical search algorithms such as the conjugate gradient search, with positive beam weight constraints, generally require numerous iterations and may produce suboptimal dose results due to trapping in local minima. A direct solution of the inverse problem using conventional quadratic objective functions without positive beam constraints is more efficient but will result in unrealistic negative beam weights. We present here a direct solution of the inverse problem that does not yield unphysical negative beam weights. The objective function for the optimization of a large number of beamlets is reformulated such that the optimization problem is reduced to a linear set of equations. The optimal set of intensities is found through a matrix inversion, and negative beamlet intensities are avoided without the need for externally imposed ad-hoc constraints. The method has been demonstrated with a test phantom and a few clinical radiotherapy cases, using primary dose calculations. We achieve highly conformal primary dose distributions with very rapid optimization times. Typical optimization times for a single anatomical slice (two dimensional) (head and neck) using a LAPACK matrix inversion routine in a single processor desktop computer, are: 0.03 s for 500 beamlets; 0.28 s for 1000 beamlets; 3.1 s for 2000 beamlets; and 12 s for 3000 beamlets. Clinical implementation will require the additional time of a one-time precomputation of scattered radiation for all beamlets, but will not impact the optimization speed. In conclusion, the new method provides a fast and robust technique to find a global minimum that yields excellent results for the inverse planning of IMRT. PMID- 16266117 TI - The impact of thermal wave characteristics on thermal dose distribution during thermal therapy: a numerical study. AB - The aim of this study was to investigate the effects of the propagation speed of a thermal wave in terms of the thermal relaxation time on the temperature/thermal dose distributions in living tissue during thermal therapies. The temperature field in tissue was solved by the finite difference method, and the thermal dose was calculated from the formulation proposed by Sapareto and Dewey [Int. J. Radiat. Oncol. Biol. Phys. 10, 787-800 (1984)]. Under the same total deposited energy, for a rapid heating process the time lagging behavior of the peak temperature became pronounced and the level of the peak temperature was decreased with increasing the thermal relaxation time. When the heating duration was longer than the thermal relaxation time of tissues, there was no significant difference between the thermal dose distributions with/without considering the effect of the thermal relaxation time. In other words, when the heating duration is comparable to or shorter than the thermal relaxation time of tissue, the results of the wave bioheat transfer equation (WBHTE) are fully different from that of the Pennes' bioheat transfer equation (PBHTE). Besides, for a rapid heating process the dimension of thermal lesion was still significantly affected by perfusion, because this is what is predicted by the WBHTE but not by the PBHTE, i.e., the wave feature of the temperature field cannot fully be predicted by the PBHTE. PMID- 16266118 TI - Real-time DMLC IMRT delivery for mobile and deforming targets. AB - In numerous cases of radiotherapy delivery to moving targets, simplifying assumptions of identical pattern of motions of tissue for each fraction are not satisfied. Therefore, algorithms capable to respond in real time to motions of target registered at treatment should be developed to improve the precision of radiation intensity delivery. The DMLC delivery of predetermined intensity maps to moving and deforming targets in real time is developed in this paper. Algorithms are constructed so that constraints on maximum admissible speed of leaves are preserved during delivery. A sequence of examples is presented to illustrate behavior of leaf trajectories for representative cases of [dynamic multileaf collimator] (DMLC) [intensity modulated radiation therapy] (IMRT) real time delivery. The examples presented show real-time deliveries to targets moving as rigid bodies and targets deforming uniformly over their volumes. Examples are admitting random perturbations of predefined target motions that are time dependent only, i.e., target motion perturbations are identical for all target points. PMID- 16266120 TI - FR258900, a novel glycogen phosphorylase inhibitor isolated from Fungus No. 138354. I. Taxonomy, fermentation, isolation and biological activities. AB - FR258900 is a novel glycogen synthesis activator produced by Fungus No. 138354. This compound was isolated from the culture broth by solvent extraction and reverse-phase column chromatography. FR258900 stimulated glycogen synthesis and glycogen synthase activity in primary rat hepatocytes. FR258900 exhibited a potent inhibitory effect on the activity of liver glycogen phosphorylase, suggesting that this compound may activate hepatic glycogen synthesis via glycogen phosphorylase inhibition. Thus, this glycogen phosphorylase inhibitor may be useful in the treatment of postprandial hyperglycemia in type 2 diabetes. PMID- 16266119 TI - Examination of dosimetry accuracy as a function of seed detection rate in permanent prostate brachytherapy. AB - The variation of permanent prostate brachytherapy dosimetry as a function of seed detection rates was investigated for I125 implants with seed activities commonly employed in contemporary practice. Post-implant imaging and radiation dosimetry data from nine patients who underwent PPB served as the basis of this simulation study. One-thousand random configurations of detected seeds were generated for each patient dataset using various seed detection levels from 30% to 99%. Dose parameters, including D90, were computed for each configuration and compared with the actual dosimetry data. A total of 108 000 complete sets of post-PPB dose volume statistics were computed. The results demonstrated that although the average D90 differed from the true value by less than 5% when 70% or more seeds were identified, the D90 of an individual case could deviate up to 13%. The 95% confidence interval (CI) of estimated D90 values differ by less than 5% from the actual value when 95% or more seeds are detected, or approximately a 7 Gy difference in the D90 value for a prescription dose of 144 Gy. Estimated target volume dose parameters tended to decrease with reduced seed detection rates. The most variable dose parameter was the prostate V100 in absolute scale while the urethral V100 was most variable in a relative sense. Based on this comprehensive simulation study, it is suggested that 95% or more seeds need to be localized in order to provide an accurate estimation of dose parameters for contemporary iodine 125 permanent prostate brachytherapy. PMID- 16266121 TI - FR258900, a novel glycogen phosphorylase inhibitor isolated from Fungus No. 138354. II. Anti-hyperglycemic effects in diabetic animal models. AB - A novel glycogen phosphorylase inhibitor FR258900 was isolated from the cultured broth of a fungal strain No. 138354. We examined the hypoglycemic effects of FR258900 in diabetic animal models. FR258900 treatment significantly reduced the plasma glucose concentrations during oral glucose tolerance tests in diabetic mice models, including db/db mice and STZ-induced diabetic mice. Furthermore, FR258900 treatment resulted in rapid decrease in the plasma glucose levels in db/db mice. These improvements in glucose disposal were accompanied by increased liver glycogen contents, suggesting that the glucose lowering effects of FR258900 were attributed to suppressed hepatic glycogen breakdown and increased hepatic glycogen synthesis. Taken together, our results suggest that glycogen phosphorylase is a potentially useful target in new therapies against diabetes. PMID- 16266122 TI - F2928-1 and -2, new antifungal antibiotics from Cladobotryum sp. AB - Two new antifungal antibiotics, F2928-1 (1) and -2 (2), were isolated from the culture broth of Cladobotryum sp. These compounds were purified by solvent extraction, silica gel column chromatography and preparative HPLC, consecutively. The structures of these compounds were assigned as a decalin compound on the basis of various spectral analyses. These compounds showed antimicrobial activity against fungi including clinically important fungus, Aspergillus fumigatus. PMID- 16266123 TI - Mycophenolic acid inhibits syncytium formation accompanied by reduction of gp120 expression. AB - Mycophenolic acid (MPA) was identified as an inhibitor of syncytium formation during the screening of human immunodeficiency virus (HIV) entry inhibitors. MPA is a well-known inhibitor of inosine monophosphate dehydrogenase and anti-HIV activity has been reported in vitro and in vivo. MPA inhibited syncytium formation in T cell-tropic and macrophage-tropic systems with IC50 values of 0.1 and 0.5 microM, respectively. The reduction of HIV gp120 expression by MPA (1.0 microM) was observed by use of Western blot analysis. Furthermore, the addition of guanosine restored both syncytium formation and gp120 expression in the presence of MPA. These results suggest that MPA inhibits not only reverse transcription by depletion of a substrate, GTP, as has been reported, but also syncytium formation through a predominant reduction in the amount of gp120 that is vigorously expressed in the above transformed cells and may be in HIV-infected cells. PMID- 16266124 TI - A new fungicide produced by a Streptomyces sp. GAAS7310. AB - Directed bioassay guided fraction led to a new macrodiolide antimycin A(17) (1), isolated from a Streptomyces sp. GAAS7310, which showed significant antimicrobial activity against eleven fungal species, including Curvularia lunata (Wakker) Boed, Rhizopus nigrtcans Ehrb and Colletotrichum nigrum EL. et Halst. The structure was unambiguously established by interpretation of 1D and 2D NMR data and comparison with the known antimycin A(1a). PMID- 16266125 TI - Absolute configuration of TPU-0043, a pentaene macrolide from Streptomyces sp. AB - An antifungal pentaene macrolide TPU-0043 was isolated from Streptomyces sp. TP A0625. The absolute configuration of TPU-0043 was determined to be 2R-(n-butyl) 16-methyl-3S,5S,7S,9R,11R,13R,15S,26S, 27R-nonahydroxyoctacosa-16,18,20,22,24 pentaenoic acid, 27-lactone, by X-ray crystallography of its 13-p bromobenzenesulfonyl derivative. PMID- 16266126 TI - 1-Hydroxy-1-norresistomycin, a new cytotoxic compound from a marine actinomycete, Streptomyces chibaensis AUBN1/7. AB - In our systematic screening programme for marine actinomycetes, a bioactive streptomycete was isolated from marine sediment samples of the Bay of Bengal, India. The isolate yielded a new cytotoxic compound. This was obtained by solvent extraction followed by chromatographic purification. The pure compound was identified from spectroscopic data as a quinone-related antibiotic, 1-hydroxy-1 norresistomycin (1). It showed a potent cytotoxic activity against cell lines viz. HMO2 (gastric adenocarcinoma) and HePG2 (hepatic carcinoma) in vitro. It also exhibited antibacterial activities against Gram-positive and Gram-negative bacteria. PMID- 16266127 TI - 1-Hydroxy-1-norresistomycin and resistoflavin methyl ether: new antibiotics from marine-derived streptomycetes. AB - Cultivation of the marine-derived streptomycete isolate B8005 delivered three known antibiotics, resistomycin (1), resistoflavin (3a) and tetracenomycin (4), and a further member of the rare resistomycin class, the weakly antibiotically active 1-hydroxy-1-norresistomycin (2). From a related marine strain B4842, 1 and resistoflavin methyl ether (3b) have been isolated. The formation of 2 is of interest from a biosynthetic point of view. PMID- 16266128 TI - Structure elucidation of Sch 725674 from Aspergillus sp. AB - A new macrolide Sch725674 (1) was isolated and identified from the culture of an Aspergillus sp. The structure elucidation of 1 was accomplished based on extensive NMR spectroscopic analyses. Compound 1 showed inhibitory activity against Saccharomyces cerevisiae (PM503) and Candida albicans (C43) with MICs of 8 and 32 microg/ml, respectively. PMID- 16266129 TI - Isolation and identification of a novel impurity of erythromycin A 9-oxime desosaminehydrazinium salt. AB - In the manufacturing process for Biaxin (clarithromycin), erythromycin-A oxime, an intermediate, is obtained in high yield, when erythromycin-A is treated with hydroxylamine/isopropyl alcohol in the presence of acetic acid. An unusual impurity, the desosamine hydrazinium salt, is generated in this step of the synthetic pathway, and has been isolated and characterized by using one and two dimensional NMR spectroscopy in conjunction with MS and EDS. PMID- 16266132 TI - Stimulus-frequency otoacoustic emissions measured with amplitude-modulated suppressor tones (L). AB - Stimulus-frequency otoacoustic emissions (SFOAEs) are typically derived as the difference in sound pressure in the ear canal with and without a suppressor tone added to the probe tone. A novel variation of this method applies a sinusoidal amplitude modulation (AM) to the suppressor tone, which causes the SFOAE to also be modulated. The AM-SFOAE can be separated from the probe frequency using spectral methods. AM-SFOAE measurements are described for four normal-hearing subjects using 6-Hz AM. Because the suppressor modulation is at a higher rate, the AM-SFOAE technique avoids the confounding influence of heartbeat, which also modulates the probe tone. PMID- 16266133 TI - The tongue stops here: ultrasound imaging of the palate (L). AB - This letter presents a method for imaging the palate and extracting the palate contour from ultrasound images. Ultrasound does not usually capture the palate because the air at the tongue surface reflects the ultrasound beam back to the transducer. However, when the tongue touches the palate during a swallow, the ultrasound beam is transmitted through the soft tissue until it reaches and is reflected by the palate. In combination with tongue contours, the palate contour has the potential for disambiguation of the tongue surface, registration of images within and across subjects, and calculation of phonetically important measures. PMID- 16266139 TI - Modifications of the equation for gas bubble dynamics in a soft elastic medium. AB - A model equation for the oscillation of a pressurized gas bubble in a nonlinear incompressible elastic medium [Emelianov et al., J. Acoust. Soc. Am. 115, 581 (2004)] is extended to include effects of surface tension, viscosity, weak compressibility, and confinement by an elastic shell. The significance of this work is that starting from first principles, the full nonlinearity of the incompressible elastic medium surrounding the bubble and forming its shell is taken into account. Measurements of equilibrium radius as a function of external pressure for a gas bubble in a tissue-like gel are also presented. A general approach to including hysteresis is also discussed. PMID- 16266143 TI - Tracking individual fish from a moving platform using a split-beam transducer. AB - Tracking of individual fish targets using a split-beam echosounder is a common method for investigating fish behavior. When mounted on a floating platform like a ship or a buoy, the transducer movement often complicates the process. This paper presents a framework for tracking single targets from such a platform. A filter based on the correlated fish movements between pings is developed to estimate the platform movement, and an extended Kalman filter is used to combine the split-beam measurements and the platform-position estimates. Different methods for gating and data association are implemented and tested with respect to data-association errors, using manually tracked data from a free-floating buoy as a reference. The data association was improved by utilizing the estimated velocity for each track to predict the location of the next observation. The data association was more robust when estimates of platform tilt/roll were used. Other techniques to estimate position and velocity, like linear regression and smoothing splines, were implemented and tested on a simulated data set. The platform-state estimation improved the estimates for methods like the Kalman filter and a smoothing spline with cross validation, but not for robust methods like linear regression and smoothing spline with a fixed degree of smoothing. PMID- 16266157 TI - Multichannel time-reversal processing for acoustic communications in a highly reverberant environment. AB - The development of time-reversal (T/R) communication systems is a recent signal processing research area dominated by applying T/R techniques to communicate in hostile environments. The fundamental concept is based on time-reversing the impulse response or Green's function characterizing the uncertain communications channel to mitigate deleterious dispersion and multipath effects. In this paper, we extend point-to-point to array-to-point communications by first establishing the basic theory to define and solve the underlying multichannel communications problem and then developing various realizations of the resulting T/R receivers. We show that not only do these receivers perform well in a hostile environment, but they also can be implemented with a "1 bit" analog-to-digital converter design structure. We validate these results by performing proof-of-principle acoustic communications experiments in air. It is shown that the resulting T/R receivers are capable of extracting the transmitted coded sequence from noisy microphone array measurements with zero-bit error. PMID- 16266160 TI - Transmission properties of bone conducted sound: measurements in cadaver heads. AB - In the past, only a few investigations have measured vibration at the cochlea with bone conduction stimulation: dry skulls were used in those investigations. In this paper, the transmission properties of bone conducted sound in human head are presented, measured as the three-dimensional vibration at the cochlear promontory in six intact cadaver heads. The stimulation was provided at 27 positions on the skull surface and two close to the cochlea; mechanical point impedance was measured at all positions. Cochlear promontory vibration levels in the three perpendicular directions were normally within 5 dB. With the stimulation applied on the ipsilateral side, the response decreased, and the accumulated phase increased, with distance between the cochlea and the excitation position. No significant changes were obtained when the excitations were on the contralateral side. In terms of vibration level, the best stimulation position is on the mastoid close to the cochlea; the worst is at the midline of the skull. The transcranial transmission was close to 0 dB for frequencies up to 700 Hz; above it decreased at 12 dB/decade. Wave transmission at the skull-base was found to be nondispersive at frequencies above 2 kHz whereas it altered with frequency at the cranial vault. PMID- 16266161 TI - Reducing individual differences in the external-ear transfer functions of the Mongolian gerbil. AB - This study examines individual differences in the directional transfer functions (DTFs), the directional components of head-related transfer functions of gerbils, and seeks a method for reducing these differences. The difference between the DTFs of a given animal pair was quantified by the intersubject spectral difference (ISSD), which is the variance in the difference spectra of DTFs for frequencies between 5 and 45 kHz and for 361 source directions. An attempt was made to reduce the ISSD by scaling the DTFs of one animal in frequency and/or rotating the DTFs along the source coordinate sphere. The ISSD was reduced by a median of 12% after optimal frequency scaling alone, by a median of 19% after optimal spatial rotation alone, and by a median of 36% after simultaneous frequency scaling and spatial rotation. The optimal scaling factor (OSF) and the optimal coordinate rotation (OCR) correlated strongly with differences in head width and pinna angles (i.e., pinna inclination around the vertical and front back axes), respectively. Thus, linear equations were derived to estimate the OSF and OCR from these anatomical measurements. The ISSD could be reduced by a median of 22% based on the estimated OSF and OCR. PMID- 16266162 TI - Comparison of an analytic horn equation approach and a boundary element method for the calculation of sound fields in the human ear canal. AB - The sound field inside a model human ear canal has been computed, to show both longitudinal variations along the canal length and transverse variations through cross-sectional slices. Two methods of computation were used. A modified horn equation approach parametrizes the sound field with a single coordinate, the position along a curved center axis-this approach can accommodate the curvature and varying cross-sectional area of the ear canal but cannot compute transverse variations of the sound field. A boundary element method (BEM) was also implemented to compute the full three-dimensional sound field. Over 2000 triangular mesh elements were used to represent the ear canal geometry. For a plane piston source at the entrance plane, the pressure along the curved center axis predicted by the two methods is in good agreement, for frequencies up to 15 kHz, for four different ear canals. The BEM approach, though, reveals spatial variations of sound pressure within each canal cross section. These variations are small below 4 kHz, but increase with frequency, reaching 1.5 dB at 8 kHz and 4.5 dB at 15 kHz. For source configurations that are more realistic than a simple piston, large transverse variations in sound pressure are anticipated in the vicinity of the source. PMID- 16266163 TI - Contribution of frequency modulation to speech recognition in noise. AB - Cochlear implants allow most patients with profound deafness to successfully communicate under optimal listening conditions. However, the amplitude modulation (AM) information provided by most implants is not sufficient for speech recognition in realistic settings where noise is typically present. This study added slowly varying frequency modulation (FM) to the existing algorithm of an implant simulation and used competing sentences to evaluate FM contributions to speech recognition in noise. Potential FM advantage was evaluated as a function of the number of spectral bands, FM depth, FM rate, and FM band distribution. Barring floor and ceiling effects, significant improvement was observed for all bands from 1 to 32 with the additional FM cue both in quiet and noise. Performance also improved with greater FM depth and rate, which might reflect resolved sidebands under the FM condition. Having FM present in low-frequency bands was more beneficial than in high-frequency bands, and only half of the bands required the presence of FM, regardless of position, to achieve performance similar to when all bands had the FM cue. These results provide insight into the relative contributions of AM and FM to speech communication and the potential advantage of incorporating FM for cochlear implant signal processing. PMID- 16266165 TI - Delays of stimulus-frequency otoacoustic emissions and cochlear vibrations contradict the theory of coherent reflection filtering. AB - When stimulated by tones, the ear appears to emit tones of its own, stimulus frequency otoacoustic emissions (SFOAEs). SFOAEs were measured in 17 chinchillas and their group delays were compared with a place map of basilar-membrane vibration group delays measured at the characteristic frequency. The map is based on Wiener-kernel analysis of responses to noise of auditory-nerve fibers corroborated by measurements of vibrations at several basilar-membrane sites. SFOAE group delays were similar to, or shorter than, basilar-membrane group delays for frequencies >4 kHz and <4 kHz, respectively. Such short delays contradict the generally accepted "theory of coherent reflection filtering" [Zweig and Shera, J. Acoust. Soc. Am. 98, 2018-2047 (1995)], which predicts that the group delays of SFOAEs evoked by low-level tones approximately equal twice the basilar-membrane group delays. The results for frequencies higher than 4 kHz are compatible with hypotheses of SFOAE propagation to the stapes via acoustic waves or fluid coupling, or via reverse basilar membrane traveling waves with speeds corresponding to the signal-front delays, rather than the group delays, of the forward waves. The results for frequencies lower than 4 kHz cannot be explained by hypotheses based on waves propagating to and from their characteristic places in the cochlea. PMID- 16266166 TI - An analysis of the impact of auditory-nerve adaptation on behavioral measures of temporal integration in cochlear implant recipients. AB - The objective of this study was to determine the impact that auditory-nerve adaptation has on behavioral measures of temporal integration in Nucleus 24 cochlear implant recipients. It was expected that, because the auditory nerve serves as the input to central temporal integrator, a large degree of auditory nerve adaptation would reduce the amount of temporal integration. Neural adaptation was measured by tracking amplitude changes of the electrically evoked compound action potential (ECAP) in response to 1000-pps biphasic pulse trains of varying durations. Temporal integration was measured at both suprathreshold and threshold levels by an adaptive procedure. Although varying degrees of neural adaptation and temporal integration were observed across individuals, results of this investigation revealed no correlation between the degree of neural adaptation and psychophysical measures of temporal integration. PMID- 16266164 TI - Medial-olivocochlear-efferent inhibition of the first peak of auditory-nerve responses: evidence for a new motion within the cochlea. AB - Despite the insights obtained from click responses, the effects of medial olivocochlear (MOC) efferents on click responses from single-auditory-nerve (AN) fibers have not been reported. We recorded responses of cat single AN fibers to randomized click level series with and without electrical stimulation of MOC efferents. MOC stimulation inhibited (1) the whole response at low sound levels, (2) the decaying part of the response at all sound levels, and (3) the first peak of the response at moderate to high sound levels. The first two effects were expected from previous reports using tones and are consistent with a MOC-induced reduction of cochlear amplification. The inhibition of the AN first peak, which was strongest in the apex and middle of the cochlea, was unexpected because the first peak of the classic basilar-membrane (BM) traveling wave receives little or no amplification. In the cochlear base, the click data were ambiguous, but tone data showed particularly short group delays in the tail-frequency region that is strongly inhibited by MOC efferents. Overall, the data support the hypothesis that there is a motion that bends inner-hair-cell stereocilia and can be inhibited by MOC efferents, a motion that is present through most, or all, of the cochlea and for which there is no counterpart in the classic BM traveling wave. PMID- 16266167 TI - Detection of high-frequency spectral notches as a function of level. AB - High-frequency spectral notches are important cues for sound localization. Our ability to detect them must depend on their representation as auditory nerve (AN) rate profiles. Because of the low threshold and the narrow dynamic range of most AN fibers, these rate profiles deteriorate at high levels. The system may compensate by using onset rate profiles whose dynamic range is wider, or by using low-spontaneous-rate fibers, whose threshold is higher. To test these hypotheses, the threshold notch depth necessary to discriminate between a flat spectrum broadband noise and a similar noise with a spectral notch centered at 8 kHz was measured at levels from 32 to 100 dB SPL. The importance of the onset rate profile representation of the notch was estimated by varying the stimulus duration and its rise time. For a large proportion of listeners, threshold notch depth varied nonmonotonically with level, increasing for levels up to 70-80 dB SPL and decreasing thereafter. The nonmonotonic aspect of the function was independent of notch bandwidth and stimulus duration. Thresholds were independent of stimulus rise time but increased for the shorter noise bursts. Results are discussed in terms of the ability of the AN to convey spectral notch information at different levels. PMID- 16266168 TI - Perceptual interaction between carrier periodicity and amplitude modulation in broadband stimuli: a comparison of the autocorrelation and modulation-filterbank model. AB - Recent temporal models of pitch and amplitude modulation perception converge on a relatively realistic implementation of cochlear processing followed by a temporal analysis of periodicity. However, for modulation perception, a modulation filterbank is applied whereas for pitch perception, autocorrelation is applied. Considering the large overlap in pitch and modulation perception, this is not parsimonious. Two experiments are presented to investigate the interaction between carrier periodicity, which produces strong pitch sensations, and envelope periodicity using broadband stimuli. Results show that in the presence of carrier periodicity, detection of amplitude modulation is impaired throughout the tested range (8-1000 Hz). On the contrary, detection of carrier periodicity in the presence of an additional amplitude modulation is impaired only for very low frequencies below the pitch range (<33 Hz). Predictions of a generic implementation of a modulation-filterbank model and an autocorrelation model are compared to the data. Both models were too insensitive to high-frequency envelope or carrier periodicity and to infra-pitch carrier periodicity. Additionally, both models simulated modulation detection quite well but underestimated the detrimental effect of carrier periodicity on modulation detection. It is suggested that a hybrid model consisting of bandpass envelope filters with a ripple in their passband may provide a functionally successful and physiologically plausible basis for a unified model of auditory periodicity extraction. PMID- 16266169 TI - Informational masking for simultaneous nonspeech stimuli: psychometric functions for fixed and randomly mixed maskers. AB - Sensitivity d' and response bias beta were measured as a function of target level for the detection of a 1000-Hz tone in multitone maskers using a one interval, two-alternative forced-choice (1I-2AFC) paradigm. Ten such maskers, each with eight randomly selected components in the region 200-5000 Hz, with 800-1250 Hz excluded to form a protected zone, were presented under two conditions: the fixed condition, in which the same eight-component masker is used throughout an experimental run, and the random condition, in which an eight-component masker is chosen randomly trial-to-trial from the given set of ten such maskers. Differences between the results obtained with these two conditions help characterize the listener's susceptibility to informational masking (IM). The d' results show great intersubject variability, but can be reasonably well fit by simple energy-detector models in which internal noise and filter bandwidth are used as fitting parameters. In contrast, the beta results are not well fit by these models. In addition to presentation of new data and its relation to energy detector models, this paper provides comments on a variety of issues, problems, and research needs in the IM area. PMID- 16266170 TI - Psychophysical tuning curves at very high frequencies. AB - For most normal-hearing listeners, absolute thresholds increase rapidly above about 16 kHz. One hypothesis is that the high-frequency limit of the hearing threshold curve is imposed by the transmission characteristics of the middle ear, which attenuates the sound input [Masterton et al., J. Acoust. Soc. Am. 45, 966 985 (1969)]. An alternative hypothesis is that the high-frequency limit of hearing is imposed by the tonotopicity of the cochlea [Ruggero and Temchin, Proc. Nat. Acad. Sci. U.S.A. 99, 13206-13210 (2002)]. The aim of this study was to test these hypotheses. Forward-masked psychophysical tuning curves (PTCs) were derived for signal frequencies of 12-17.5 kHz. For the highest signal frequencies, the high-frequency slopes of some PTCs were steeper than the slope of the hearing threshold curve. The results also show that the human auditory system displays frequency selectivity for characteristic frequencies (CFs) as high as 17 kHz, above the frequency at which absolute thresholds begin to increase rapidly. The findings suggest that, for CFs up to 17 kHz, the high-frequency limitation in humans is imposed in part by the middle-ear attenuation, and not by the tonotopicity of the cochlea. PMID- 16266172 TI - Consequences of cochlear damage for the detection of interaural phase differences. AB - Thresholds for detecting interaural phase differences (IPDs) in sinusoidally amplitude-modulated pure tones were measured in seven normal-hearing listeners and nine listeners with bilaterally symmetric hearing losses of cochlear origin. The IPDs were imposed either on the carrier signal alone-not the amplitude modulation-or vice versa. The carrier frequency was 250, 500, or 1000 Hz, the modulation frequency 20 or 50 Hz, and the sound pressure level was fixed at 75 dB. A three-interval two-alternative forced choice paradigm was used. For each type of IPD (carrier or modulation), thresholds were on average higher for the hearing-impaired than for the normal listeners. However, the impaired listeners' detection deficit was markedly larger for carrier IPDs than for modulation IPDs. This was not predictable from the effect of hearing loss on the sensation level of the stimuli since, for normal listeners, large reductions of sensation level appeared to be more deleterious to the detection of modulation IPDs than to the detection of carrier IPDs. The results support the idea that one consequence of cochlear damage is a deterioration in the perceptual sensitivity to the temporal fine structure of sounds. PMID- 16266171 TI - Monaural and interaural temporal modulation transfer functions measured with 5 kHz carriers. AB - Temporal modulation transfer functions (TMTFs) were measured for detection of monaural sinusoidal amplitude modulation and dynamically varying interaural level differences for a single set of listeners. For the interaural TMTFs, thresholds are the modulation depths at which listeners can just discriminate interaural envelope-phase differences of 0 and 180 degrees. A 5-kHz pure tone and narrowband noises, 30- and 300-Hz wide centered at 5 kHz, were used as carriers. In the interaural conditions, the noise carriers were either diotic or interaurally uncorrelated. The interaural TMTFs with tonal and diotic noise carriers exhibited a low-pass characteristic but the cutoff frequencies changed nonmonotonically with increasing bandwidth. The interaural TMTFs for the tonal carrier began rolling off approximately a half-octave lower than the tonal monaural TMTF (approximately 80 Hz vs approximately 120 Hz). Monaural TMTFs obtained with noise carriers showed effects attributable to masking of the signal modulation by intrinsic fluctuations of the carrier. In the interaural task with dichotic noise carriers, similar masking due to the interaural carrier fluctuations was observed. Although the mechanisms responsible for differences between the monaural and interaural TMTFs are unknown, the lower binaural TMTF cutoff frequency suggests that binaural processing exhibits greater temporal limitation than monaural processing. PMID- 16266173 TI - Tactile information transfer: a comparison of two stimulation sites. AB - Two experiments on the discrimination of time-varying tactile stimuli were performed, with comparison of stimulus delivery to the distal pad of the right index finger and to the right wrist (palmar surface). Subjects were required to perceive differences in short sequences of computer-generated stimulus elements (experiment 1) or differences in short tactile stimuli derived from a speech signal (experiment 2). The pulse-train stimuli were distinguished by differences in frequency (i.e., pulse repetition rate) and amplitude, and by the presence/absence of gaps (approximately 100-ms duration). Stimulation levels were 10 dB higher at the wrist than at the fingertip, to compensate for the lower vibration sensitivity at the wrist. Results indicate similar gap detection at wrist and fingertip and similar perception of frequency differences. However, perception of amplitude differences was found to be better at the wrist than at the fingertip. Maximum information transfer rates for the stimuli in experiment 1 were estimated at 7 bits s(-1) at the wrist and 5 bits s(-1) at the fingertip. PMID- 16266175 TI - Comparison of nonlinear dynamic methods and perturbation methods for voice analysis. AB - Nonlinear dynamic methods and perturbation methods are compared in terms of the effects of signal length, sampling rate, and noise. Results of theoretical and experimental studies quantitatively show that measurements representing frequency and amplitude perturbations are not applicable to chaotic signals because of difficulties in pitch tracking and sensitivity to initial state differences. Perturbation analyses are only reliable when applied to nearly periodic voice samples of sufficiently long signal lengths that were obtained at high sampling rates and low noise levels. In contrast, nonlinear dynamic methods, such as correlation dimension, allow the quantification of chaotic time series. Additionally, the correlation dimension method presents a more stable analysis of nearly periodic voice samples for shorter signal lengths, lower sampling rates, and higher noise levels. The correlation dimension method avoids some of the methodological issues associated with perturbation methods, and may potentially improve the ability for real time analysis as well as reduce costs in experimental designs for objectively assessing voice disorders. PMID- 16266174 TI - Multidimensional analyses of voicing offsets and onsets in female speakers. AB - This study investigates cross-speaker differences in the factors that predict voicing thresholds during abduction-adduction gestures in six normal women. Measures of baseline airflow, pulse amplitude, subglottal pressure, and fundamental frequency were made at voicing offset and onset during intervocalic /h/, produced in varying vowel environments and at different loudness levels, and subjected to relational analyses to determine which factors were most strongly related to the timing of voicing cessation or initiation. The data indicate that (a) all speakers showed differences between voicing offsets and onsets, but the degree of this effect varied across speakers; (b) loudness and vowel environment have speaker-specific effects on the likelihood of devoicing during /h/; and (c) baseline flow measures significantly predicted times of voicing offset and onset in all participants, but other variables contributing to voice timing differed across speakers. Overall, the results suggest that individual speakers have unique methods of achieving phonatory goals during running speech. These data contribute to the literature on individual differences in laryngeal function, and serve as a means of evaluating how well laryngeal models can reproduce the range of voicing behavior used by speakers during running speech tasks. PMID- 16266176 TI - Lexical frequency and acoustic reduction in spoken Dutch. AB - This study investigates the effects of lexical frequency on the durational reduction of morphologically complex words in spoken Dutch. The hypothesis that high-frequency words are more reduced than low-frequency words was tested by comparing the durations of affixes occurring in different carrier words. Four Dutch affixes were investigated, each occurring in a large number of words with different frequencies. The materials came from a large database of face-to-face conversations. For each word containing a target affix, one token was randomly selected for acoustic analysis. Measurements were made of the duration of the affix as a whole and the durations of the individual segments in the affix. For three of the four affixes, a higher frequency of the carrier word led to shorter realizations of the affix as a whole, individual segments in the affix, or both. Other relevant factors were the sex and age of the speaker, segmental context, and speech rate. To accommodate for these findings, models of speech production should allow word frequency to affect the acoustic realizations of lower-level units, such as individual speech sounds occurring in affixes. PMID- 16266177 TI - Acoustic and spectral characteristics of young children's fricative productions: a developmental perspective. AB - Scientists have made great strides toward understanding the mechanisms of speech production and perception. However, the complex relationships between the acoustic structures of speech and the resulting psychological percepts have yet to be fully and adequately explained, especially in speech produced by younger children. Thus, this study examined the acoustic structure of voiceless fricatives (/f, theta, s, S/) produced by adults and typically developing children from 3 to 6 years of age in terms of multiple acoustic parameters (durations, normalized amplitude, spectral slope, and spectral moments). It was found that the acoustic parameters of spectral slope and variance (commonly excluded from previous studies of child speech) were important acoustic parameters in the differentiation and classification of the voiceless fricatives, with spectral variance being the only measure to separate all four places of articulation. It was further shown that the sibilant contrast between /s/ and /S/ was less distinguished in children than adults, characterized by a dramatic change in several spectral parameters at approximately five years of age. Discriminant analysis revealed evidence that classification models based on adult data were sensitive to these spectral differences in the five-year-old age group. PMID- 16266178 TI - Finding intonational boundaries using acoustic cues related to the voice source. AB - Acoustic cues related to the voice source, including harmonic structure and spectral tilt, were examined for relevance to prosodic boundary detection. The measurements considered here comprise five categories: duration, pitch, harmonic structure, spectral tilt, and amplitude. Distributions of the measurements and statistical analysis show that the measurements may be used to differentiate between prosodic categories. Detection experiments on the Boston University Radio Speech Corpus show equal error detection rates around 70% for accent and boundary detection, using only the acoustic measurements described, without any lexical or syntactic information. Further investigation of the detection results shows that duration and amplitude measurements, and, to a lesser degree, pitch measurements, are useful for detecting accents, while all voice source measurements except pitch measurements are useful for boundary detection. PMID- 16266179 TI - Effects of cochlear hearing loss on perceptual grouping cues in competing-vowel perception. AB - This study compared how normal-hearing listeners (NH) and listeners with moderate to moderately severe cochlear hearing loss (HI) use and combine information within and across frequency regions in the perceptual separation of competing vowels with fundamental frequency differences (deltaF0) ranging from 0 to 9 semitones. Following the procedure of Culling and Darwin [J. Acoust. Soc. Am. 93, 3454-3467 (1993)], eight NH listeners and eight HI listeners identified competing vowels with either a consistent or inconsistent harmonic structure. Vowels were amplified to assure audibility for HI listeners. The contribution of frequency region depended on the value of deltaF0 between the competing vowels. When deltaF0 was small, both groups of listeners effectively utilized deltaF0 cues in the low-frequency region. In contrast, HI listeners derived significantly less benefit than NH listeners from deltaF0 cues conveyed by the high-frequency region at small deltaF0's. At larger deltaF0's, both groups combined deltaF0 cues from the low and high formant-frequency regions. Cochlear impairment appears to negatively impact the ability to use F0 cues for within-formant grouping in the high-frequency region. However, cochlear loss does not appear to disrupt the ability to use within-formant F0 cues in the low-frequency region or to group F0 cues across formant regions. PMID- 16266180 TI - Pattern playback revisited: unvoiced stop consonant perception. AB - Among the most influential publications in speech perception is Liberman, Delattre, and Cooper's [Am. J. Phys. 65, 497-516 (1952)] report on the identification of synthetic, voiceless stops generated by the Pattern Playback. Their map of stop consonant identification shows a highly complex relationship between acoustics and perception. This complex mapping poses a challenge to many classes of relatively simple pattern recognition models which are unable to capture the original finding of Liberman et al. that identification of /k/ was bimodal for bursts preceding front vowels but otherwise unimodal. A replication of this experiment was conducted in an attempt to reproduce these identification patterns using a simulation of the Pattern Playback device. Examination of spectrographic data from stimuli generated by the Pattern Playback revealed additional spectral peaks that are consistent with harmonic distortion characteristic of tube amplifiers of that era. Only when harmonic distortion was introduced did bimodal /k/ responses in front-vowel context emerge. The acoustic consequence of this distortion is to add, e.g., a high-frequency peak to midfrequency bursts or a midfrequency peak to a low-frequency burst. This likely resulted in additional /k/ responses when the second peak approximated the second formant of front vowels. Although these results do not challenge the main observations made by Liberman et al. that perception of stop bursts is context dependent, they do show that the mapping from acoustics to perception is much less complex without these additional distortion products. PMID- 16266181 TI - Phonetic identification in quiet and in noise by listeners with cochlear implants. AB - This study examined the effect of noise on the identification of four synthetic speech continua (/ra/-/la/, /wa/-/ja/, /i/-/u/, and say-stay) by adults with cochlea implants (CIs) and adults with normal-hearing (NH) sensitivity in quiet and noise. Significant group-by-SNR interactions were found for endpoint identification accuracy for all continua except /i/-/u/. The CI listeners showed the least NH-like identification functions for the /ra/-/la/ and /wa/-/ja/ continua. In a second experiment, NH adults identified four- and eight-band cochlear implant stimulations of the four continua, to examine whether group differences in frequency selectivity could account for the group differences in the first experiment. Number of bands and SNR interacted significantly for /ra/ /la/, /wa/-/ja/, and say-stay endpoint identification; strongest effects were found for the /ra/-/la/ and say-stay continua. Results suggest that the speech features that are most vulnerable to misperception in noise by listeners with CIs are those whose acoustic cues are rapidly changing spectral patterns, like the formant transitions in the /wa/-/ja/ and /ra/-/la/ continua. However, the group differences in the first experiment cannot be wholly attributable to frequency selectivity differences, as the number of bands in the second experiment affected performance differently than suggested by group differences in the first experiment. PMID- 16266182 TI - Incidental categorization of spectrally complex non-invariant auditory stimuli in a computer game task. AB - This study examined perceptual learning of spectrally complex nonspeech auditory categories in an interactive multi-modal training paradigm. Participants played a computer game in which they navigated through a three-dimensional space while responding to animated characters encountered along the way. Characters' appearances in the game correlated with distinctive sound category distributions, exemplars of which repeated each time the characters were encountered. As the game progressed, the speed and difficulty of required tasks increased and characters became harder to identify visually, so quick identification of approaching characters by sound patterns was, although never required or encouraged, of gradually increasing benefit. After 30 min of play, participants performed a categorization task, matching sounds to characters. Despite not being informed of audio-visual correlations, participants exhibited reliable learning of these patterns at posttest. Categorization accuracy was related to several measures of game performance and category learning was sensitive to category distribution differences modeling acoustic structures of speech categories. Category knowledge resulting from the game was qualitatively different from that gained from an explicit unsupervised categorization task involving the same stimuli. Results are discussed with respect to information sources and mechanisms involved in acquiring complex, context-dependent auditory categories, including phonetic categories, and to multi-modal statistical learning. PMID- 16266183 TI - Robust acoustic object detection. AB - We consider a novel approach to the problem of detecting phonological objects like phonemes, syllables, or words, directly from the speech signal. We begin by defining local features in the time-frequency plane with built in robustness to intensity variations and time warping. Global templates of phonological objects correspond to the coincidence in time and frequency of patterns of the local features. These global templates are constructed by using the statistics of the local features in a principled way. The templates have clear phonetic interpretability, are easily adaptable, have built in invariances, and display considerable robustness in the face of additive noise and clutter from competing speakers. We provide a detailed evaluation of the performance of some diphone detectors and a word detector based on this approach. We also perform some phonetic classification experiments based on the edge-based features suggested here. PMID- 16266184 TI - Vibrato of saxophones. AB - Several alto saxophone players' vibratos have been recorded. The signals are analyzed using time-frequency methods in order to estimate the frequency modulation (vibrato rate) and the amplitude modulation (vibrato extent) of each vibrato sample. Some parameters are derived from the results in order to separate the two ways of vibrato playing: vibrato "a la machoire" and vibrato "sur l'air." Moreover, time domain simulations of single-reed instrument vibratos are created. The model is controlled by two parameters: the mouth overpressure and a parameter characterizing the reed-mouthpiece system. Preliminary comments and comparisons between the simulated vibratos and recorded vibratos results are made. PMID- 16266185 TI - Study of the brightness of trumpet tones. AB - This study focuses on a particular attribute of trumpet tones, the brightness, and on the physical characteristics of the instrument thought to govern its magnitude. On the one hand, an objective study was carried out with input impedance measurements, and, on the other hand, a subjective study with hearing tests and a panel of subjects. To create a set of different trumpets a variable depth mouthpiece was developed whose depth can be easily and continuously adjusted from "deep" to "shallow." Using this mouthpiece and the same trumpet, several instruments were generated which may be played in three ways: (i) by a musician, (ii) by an artificial mouth, and (iii) using physical modeling simulations. The influence of the depth of the mouthpiece on the perception of the trumpet's tones was investigated, and the ability of a musician, the artificial mouth, or physical modeling simulations to demonstrate perceptively noticeable differences was assessed. Physical characteristics extracted from the impedance curves are finally proposed to explain the brightness of trumpet tones. As a result, the physical modeling simulations now seem to be mature enough to exhibit coherent and subtle perceptual differences between tones. This opens the door to virtual acoustics for instrument makers. PMID- 16266186 TI - Modeling elastic wave propagation in kidney stones with application to shock wave lithotripsy. AB - A time-domain finite-difference solution to the equations of linear elasticity was used to model the propagation of lithotripsy waves in kidney stones. The model was used to determine the loading on the stone (principal stresses and strains and maximum shear stresses and strains) due to the impact of lithotripsy shock waves. The simulations show that the peak loading induced in kidney stones is generated by constructive interference from shear waves launched from the outer edge of the stone with other waves in the stone. Notably the shear wave induced loads were significantly larger than the loads generated by the classic Hopkinson or spall effect. For simulations where the diameter of the focal spot of the lithotripter was smaller than that of the stone the loading decreased by more than 50%. The constructive interference was also sensitive to shock rise time and it was found that the peak tensile stress reduced by 30% as rise time increased from 25 to 150 ns. These results demonstrate that shear waves likely play a critical role in stone comminution and that lithotripters with large focal widths and short rise times should be effective at generating high stresses inside kidney stones. PMID- 16266187 TI - Male sperm whale acoustic behavior observed from multipaths at a single hydrophone. AB - Sperm whales generate transient sounds (clicks) when foraging. These clicks have been described as echolocation sounds, a result of having measured the source level and the directionality of these signals and having extrapolated results from biosonar tests made on some small odontocetes. The authors propose a passive acoustic technique requiring only one hydrophone to investigate the acoustic behavior of free-ranging sperm whales. They estimate whale pitch angles from the multipath distribution of click energy. They emphasize the close bond between the sperm whale's physical and acoustic activity, leading to the hypothesis that sperm whales might, like some small odontocetes, control click level and rhythm. An echolocation model estimating the range of the sperm whale's targets from the interclick interval is computed and tested during different stages of the whale's dive. Such a hypothesis on the echolocation process would indicate that sperm whales echolocate their prey layer when initiating their dives and follow a methodic technique when foraging. PMID- 16266188 TI - Behavioral and auditory evoked potential audiograms of a false killer whale (Pseudorca crassidens). AB - Behavioral and auditory evoked potential (AEP) audiograms of a false killer whale were measured using the same subject and experimental conditions. The objective was to compare and assess the correspondence of auditory thresholds collected by behavioral and electrophysiological techniques. Behavioral audiograms used 3-s pure-tone stimuli from 4 to 45 kHz, and were conducted with a go/no-go modified staircase procedure. AEP audiograms used 20-ms sinusoidally amplitude-modulated tone bursts from 4 to 45 kHz, and the electrophysiological responses were received through gold disc electrodes in rubber suction cups. The behavioral data were reliable and repeatable, with the region of best sensitivity between 16 and 24 kHz and peak sensitivity at 20 kHz. The AEP audiograms produced thresholds that were also consistent over time, with range of best sensitivity from 16 to 22.5 kHz and peak sensitivity at 22.5 kHz. Behavioral thresholds were always lower than AEP thresholds. However, AEP audiograms were completed in a shorter amount of time with minimum participation from the animal. These data indicated that behavioral and AEP techniques can be used successfully and interchangeably to measure cetacean hearing sensitivity. PMID- 16266189 TI - Temporary threshold shift in bottlenose dolphins (Tursiops truncatus) exposed to mid-frequency tones. AB - A behavioral response paradigm was used to measure hearing thresholds in bottlenose dolphins before and after exposure to 3 kHz tones with sound exposure levels (SELs) from 100 to 203 dB re 1 microPa2 s. Experiments were conducted in a relatively quiet pool with ambient noise levels below 55 dB re 1 microPa2/Hz at frequencies above 1 kHz. Experiments 1 and 2 featured 1-s exposures with hearing tested at 4.5 and 3 kHz, respectively. Experiment 3 featured 2-, 4-, and 8-s exposures with hearing tested at 4.5 kHz. For experiment 2, there were no significant differences between control and exposure sessions. For experiments 1 and 3, exposures with SEL=197 dB re 1 microPa2 s and SEL > or = 195 dB re 1 microPa2 s, respectively, resulted in significantly higher TTS4 than control sessions. For experiment 3 at SEL= 195 dB re 1 microPa2 s, the mean TTS4 was 2.8 dB. These data are consistent with prior studies of TTS in dolphins exposed to pure tones and octave band noise and suggest that a SEL of 195 dB re 1 microPa2 s is a reasonable threshold for the onset of TTS in dolphins and white whales exposed to midfrequency tones. PMID- 16266190 TI - Alarm signals of the great gerbil: acoustic variation by predator context, sex, age, individual, and family group. AB - The great gerbil, Rhombomys opinus, is a highly social rodent that usually lives in family groups consisting of related females, their offspring, and an adult male. The gerbils emit alarm vocalizations in the presence of diverse predators with different hunting tactics. Alarm calls were recorded in response to three predators, a monitor lizard, hunting dog, and human, to determine whether the most common call type, the rhythmic call, is functionally referential with regard to type of predator. Results show variation in the alarm calls of both adults and subadults with the type of predator. Discriminant function analysis classified an average of 70% of calls to predator type. Call variation, however, was not limited to the predator context, because signal structure also differed by sex, age, individual callers, and family groups. These variations illustrate the flexibility of the rhythmic alarm call of the great gerbil and how it might have multiple functions and communicate in multiple contexts. Three alarm calls, variation in the rhythmic call, and vibrational signals generated from foot drumming provide the gerbils with a varied and multi-channel acoustic repertoire. PMID- 16266191 TI - Bulk ablation of soft tissue with intense ultrasound: modeling and experiments. AB - Methods for the bulk ablation of soft tissue using intense ultrasound, with potential applications in the thermal treatment of focal tumors, are presented. An approximate analytic model for bulk ablation predicts the progress of ablation based on tissue properties, spatially averaged ultrasonic heat deposition, and perfusion. The approximate model allows the prediction of threshold acoustic powers required for ablation in vivo as well as the comparison of cases with different starting temperatures and perfusion characteristics, such as typical in vivo and ex vivo experiments. In a full three-dimensional numerical model, heat deposition from array transducers is computed using the Fresnel approximation and heat transfer in tissue is computed by finite differences, accounting for heating changes caused by boiling and thermal dose-dependent absorption. Similar ablation trends due to perfusion effects are predicted by both the simple analytic model and the full numerical model. Comparisons with experimental results show the efficacy of both models in predicting tissue ablation effects. Phenomena illustrated by the simulations and experiments include power thresholds for in vivo ablation, differences between in vivo and ex vivo lesioning for comparable source conditions, the effect of tissue boiling and absorption changes on ablation depth, and the performance of a continuous rotational scanning method suitable for interstitial bulk ablation of soft tissue. PMID- 16266192 TI - Whistles of small groups of Sotalia fluviatilis during foraging behavior in southeastern Brazil. AB - Whistle emissions were recorded from small groups of marine tucuxi dolphins (Sotalia fluviatilis) in two beaches located in an important biological reserve in the Cananeia estuary (25 degrees 03'S, 47 degrees 58'W), southeastern Brazil. A total of 17 h of acoustic data was collected when dolphins were engaged in a specific feeding foraging activity. The amount of 3235 whistles was recorded and 40% (n=1294) were analyzed. Seven acoustic whistle parameters were determined: duration (ms), number of inflection points, start and end frequency (kHz), minimum and maximum frequency (kHz), and frequency range (kHz). Whistles with up to four inflection points were found. Whistles with no inflection points and rising frequency corresponded to 85% (n=1104) of all analyzed whistles. Whistle duration varied from 38 to 627 ms (mean=229.6+/-109.9 ms), with the start frequency varying between 1 and 16 kHz (mean=8.16+/-3.0 kHz) and the end frequency between 2 and 18 kHz (mean=14.35+/-3.0 kHz). The importance of this study requires an accurate measurement of the whistles' emissions in an unusual foraging feeding behavior situation on two beaches where several tucuxis, mostly mother-calf pairs, are frequently present. These two beaches are located in a federal and state environment Environmental Protected Area threatened by the progressive increase of tourism. PMID- 16266193 TI - Treatment for erectile dysfunction based on patient-reported outcomes: to every man the PDE5 inhibitor that he finds superior. AB - Erectile dysfunction (ED) is a common medical condition linked both to aging and to many medical conditions such as diabetes mellitus and cardiovascular disease. Although a common condition, treatment for ED has in the past been conducted by a few specialists, mostly urologists and sex therapists. The revolutionary introduction of oral therapy, and the massive amount of research into sexual dysfunction that followed, has led to paradigm shift in the treatment of ED. This is no longer something done by a few for a few; it involves all disciplines of medicine and more patients are being treated by a greater number of physicians. Several medications administered by different routes are available for treating ED but oral pharmacotherapy represents the first-line option. Phosphodiesterase (PDE) type 5 inhibitors are the most widely prescribed oral agents and they have a satisfactory efficacy-safety profile in patients of all categories. An alternative for men who do not respond to PDE5 inhibitors is intracavernosal injection therapy with alprostadil, a prostaglandin analogue. Other alternatives include sublingual apomorphine and intraurethral alprostadil. Both agents have a less satisfactory efficacy profile than PDE5 inhibitors and a low compliance rate. The aim of ED treatment is to restore an erection satisfactory for the sexual needs of the patient. Thus, the patient-reported outcome is the gold standard in efficacy evaluation. There are now three PDE inhibitors available, all with satisfactory efficacy-safety profiles, but with different pharmacokinetic properties. The availability of three different agents has initiated studies aiming to evaluate them regarding patient preference. However, the results are rather conflicting with some studies suggesting that tadalafil has the best patient preference, while others fail to demonstrate a clinically significant difference between the three agents. However, there is a tendency for younger men to choose tadalafil because it gives them a broader window of opportunity, while older men tend to prefer vardenafil or sildenafil. These data could be used when making a decision on which PDE5 inhibitor to prescribe, although another option is to let the patient try all three available agents and make his own choice. PMID- 16266195 TI - Docetaxel in hormone-refractory metastatic prostate cancer. AB - The taxoid analogue docetaxel is a potent inhibitor of microtubular depolymerisation and, in hormone-refractory metastatic prostate cancer, it also counters the effects of the anti-apoptotic protein Bcl-2. Overall survival was significantly increased in patients with hormone-refractory metastatic prostate cancer receiving intravenous docetaxel every 3 weeks plus oral prednisone or estramustine, compared with patients receiving intravenous mitoxantrone every 3 weeks plus prednisone in two large phase III trials (TAX 327 and SWOG [Southwest Oncology Group] 9916). In the TAX 327 study, patients receiving docetaxel 75 mg/m(2) every 3 weeks plus prednisone had a median overall survival duration of 18.9 months; in the SWOG 9916 study, median overall survival duration was 17.5 months with docetaxel 60 mg/m(2) every 3 weeks plus estramustine 280 mg three times daily on days 1-5. The median overall survival duration for the control arm of mitoxantrone 12 mg/m(2) every 3 weeks plus prednisone was 16-17 months. Compared with mitoxantrone plus prednisone, docetaxel plus prednisone improved prostate specific antigen response rate, pain and health-related quality of life, and docetaxel plus estramustine increased progression-free survival. Adverse events were more common with docetaxel- than mitoxantrone-based treatment regimens, but most events associated with docetaxel were mild-to-moderate in severity. PMID- 16266199 TI - Low-dose ethinylestradiol/levonorgestrel. AB - Low-dose ethinylestradiol/levonorgestrel 20 microg/100 microg is a combined oral contraceptive that prevents pregnancy primarily by inhibiting ovulation. The Pearl index (pregnancies per 100 woman-years of use) with ethinylestradiol/levonorgestrel 20 microg/100 microg was 0.88 and the cumulative pregnancy rate was 1.9% at the end of a 3-year open-label trial (1708 women with 26 554 evaluable cycles). The contraceptive efficacy of ethinylestradiol/levonorgestrel 20 microg/100 microg was similar to that of other low-dose combined oral contraceptives containing ethinylestradiol 20 or 35 microg in a 6-cycle trial (463 evaluable women). Ethinylestradiol/levonorgestrel 20 microg/100 microg is well tolerated; adverse events were those commonly associated with combined oral contraceptives. Headache and metrorrhagia (2% of women) were the most common adverse events leading to treatment discontinuation in the 3-year trial. Cycle control in open-label trials in women receiving up to 36 cycles of ethinylestradiol/levonorgestrel 20 microg/100 microg was generally good, with the incidence of intermenstrual bleeding being highest during the first few cycles of use and decreasing thereafter. PMID- 16266194 TI - Biologic therapy for inflammatory bowel disease. AB - Despite all of the advances in our understanding of the pathophysiology of inflammatory bowel disease (IBD), we still do not know its cause. Some of the most recently available data are discussed in this review; however, this field is changing rapidly and it is increasingly becoming accepted that immunogenetics play an important role in the predisposition, modulation and perpetuation of IBD. The role of intestinal milieu, and enteric flora in particular, appears to be of greater significance than previously thought. This complex interplay of genetic, microbial and environmental factors culminates in a sustained activation of the mucosal immune and non-immune response, probably facilitated by defects in the intestinal epithelial barrier and mucosal immune system, resulting in active inflammation and tissue destruction. Under normal situations, the intestinal mucosa is in a state of 'controlled' inflammation regulated by a delicate balance of proinflammatory (tumour necrosis factor [TNF]-alpha, interferon [IFN]-gamma, interleukin [IL]-1, IL-6, IL-12) and anti-inflammatory cytokines (IL-4, IL-10, IL 11). The mucosal immune system is the central effector of intestinal inflammation and injury, with cytokines playing a central role in modulating inflammation. Cytokines may, therefore, be a logical target for IBD therapy using specific cytokine inhibitors. Biotechnology agents targeted against TNF, leukocyte adhesion, T-helper cell (T(h))-1 polarisation, T-cell activation or nuclear factor (NF)-kappaB, and other miscellaneous therapies are being evaluated as potential therapies for IBD. In this context, infliximab is currently the only biologic agent approved for the treatment of inflammatory and fistulising Crohn's disease. Other anti-TNF biologic agents have emerged, including CDP 571, certolizumab pegol (CDP 870), etanercept, onercept and adalimumab. However, ongoing research continues to generate new biologic agents targeted at specific pathogenic mechanisms involved in the inflammatory process. Lymphocyte endothelial interactions mediated by adhesion molecules are important in leukocyte migration and recruitment to sites of inflammation, and selective blockade of these adhesion molecules is a novel and promising strategy to treat Crohn's disease. Therapeutic agents that inhibit leukocyte trafficking include natalizumab, MLN-02 and alicaforsen (ISIS 2302). Other agents being investigated for the treatment of Crohn's disease include inhibitors of T-cell activation, peroxisome proliferator-activated receptors, proinflammatory cytokine receptors and T(h)1 polarisation, and growth hormone and growth factors. Agents being investigated for treatment of ulcerative colitis include many of those mentioned for Crohn's disease. More controlled clinical trials are currently being conducted, exploring the safety and efficacy of old and new biologic agents, and the search certainly will open new and exciting perspectives on the development of therapies for IBD. PMID- 16266202 TI - Atazanavir: a review of its use in the management of HIV infection. AB - Atazanavir (Reyataz) is a novel protease inhibitor (PI) approved for use in combination with other antiretroviral drugs for the treatment of HIV infection. In antiretroviral therapy (ART)-experienced patients the drug is administered with low-dose ritonavir (i.e. boosted). In the US, unboosted atazanavir is also approved for use in ART-naive patients. In adult patients with HIV infection, atazanavir-containing highly active antiretroviral therapy (HAART) regimens provided marked improvements in virological and immunological markers and was generally well tolerated. Furthermore, recommended atazanavir regimens were no less effective than, and generally as well tolerated as, other HAART regimens in these patients, including regimens containing co-formulated lopinavir/ritonavir. Atazanavir may have an advantage over other PIs because of its favourable effect on lipid profiles, once-daily dosing, low capsule burden and, in patients with low prior PI exposure, a favourable resistance profile. Given these advantages and taking into consideration between-country differences in the approved indications, atazanavir is a valuable option as the PI component of HAART for the management of HIV infection in adult ART-naive patients, particularly where metabolic complications are a concern, and as a first- or second-line PI in combination with low-dose ritonavir in adult ART-experienced patients. PMID- 16266203 TI - Oxycodone/Ibuprofen combination tablet: a review of its use in the management of acute pain. AB - Oxycodone/ibuprofen 5 mg/400 mg (Combunox) is an oral fixed-dose combination tablet with analgesic, anti-inflammatory and antipyretic properties. It is approved in the US for the short-term (up to 7 days) management of acute, moderate-to-severe pain and is the first and only fixed-dose combination containing ibuprofen and oxycodone. A single dose of oxycodone/ibuprofen 5 mg/400 mg provided better analgesia than low-dose oxycodone or ibuprofen administered alone in most trials and appears to be more effective than a single dose of some other fixed-dose combination analgesics. It is generally well tolerated after single or multiple doses and short-term use is not expected to produce any of the serious adverse effects typically associated with the long-term use of opioids or NSAIDs. Thus, oxycodone/ibuprofen 5 mg/400mg is an effective, convenient treatment option for the short-term management of acute, moderate-to-severe pain. PMID- 16266204 TI - Eprosartan: a review of its use in the management of hypertension. AB - The angiotensin II receptor antagonist eprosartan is approved for the treatment of essential hypertension and may be administered using a convenient once-daily regimen. The drug is a well tolerated and effective antihypertensive agent with benefit in the secondary prevention of cerebrovascular events, independent of blood pressure (BP)-lowering effects. Eprosartan has a low potential for serious adverse events and has not been associated with clinically significant drug interactions, establishing it as a promising agent for combination antihypertensive strategies. Unlike ACE inhibitors such as enalapril, eprosartan does not have a tendency to cause persistent nonproductive cough. Accordingly, eprosartan represents a useful therapeutic option in the management of patients with hypertension, including those who have had a stroke and those with co-morbid type 2 diabetes mellitus. PMID- 16266207 TI - Resting metabolic rate is not reduced in obese adults with Down syndrome. AB - Resting metabolic rate (RMR) of 22 individuals with Down syndrome was compared to that of 20 nondisabled control individuals of similar age (25.7 and 27.4 years, respectively). Using a ventilated hood system, we measured RMR in the early morning after an overnight fast. Peak aerobic capacity (VO2peak) and body composition were also determined. Resting metabolic rate was not different between groups. Adjusting RMR for body weight or body surface area did not change these findings. Using stepwise regression for the total population and each subgroup, we found that only body surface area was a significant predictor of RMR. These data show that individuals with Down syndrome do not have lower RMR than their nondisabled peers, suggesting that reduced RMR does not explain the high incidence of obesity in this population. PMID- 16266205 TI - Escitalopram: a review of its use in the management of major depressive disorder. AB - Escitalopram (Cipralex, Lexapro), the active S-enantiomer of the racemic selective serotonin reuptake inhibitor (SSRI) citalopram (RS-citalopram), is a highly selective inhibitor of the serotonin transporter protein. It possesses a rapid onset of antidepressant activity, and is an effective and generally well tolerated treatment for moderate-to-severe major depressive disorder (MDD). Pooled analyses from an extensive clinical trial database suggest that escitalopram is consistently more effective than citalopram in moderate-to-severe MDD. Preliminary studies suggest that escitalopram is as effective as other SSRIs and the extended-release (XR) formulation of the serotonin/noradrenaline (norepinephrine) reuptake inhibitor venlafaxine, and may have cost-effectiveness and cost-utility advantages. However, additional longer-term, comparative studies evaluating specific efficacy, tolerability, health-related quality of life and economic indices would be helpful in definitively positioning escitalopram relative to these other agents in the treatment of MDD. Nevertheless, available clinical and pharmacoeconomic data indicate that escitalopram is an effective first-line option in the management of patients with MDD. PMID- 16266208 TI - Impact of information and weekly contact on attitudes of Korean general educators and nondisabled students regarding peers with disabilities. AB - Effects of written information and weekly special educator contact on general educators in Seoul, Korea was examined. Special educators prepared a weekly newsletter to educate teachers on how to include children with disabilities in their classrooms for part of the day. Participants were 30 general educators and 300 nondisabled peers of the students with disabilities. A pretest-posttest control group was used to assess teacher efficacy, attitudes of general educators toward inclusion, and peer acceptance of children with disabilities. We conducted ANCOVAS on posttest scores of three measures, considering pretest scores of each survey as the covariate. General educators in the information group showed significantly higher scores in teacher-efficacy and attitudes toward inclusion. Their nondisabled students showed significantly higher acceptance scores than did controls. PMID- 16266206 TI - Gemtuzumab ozogamicin: a review of its use in acute myeloid leukaemia. AB - Gemtuzumab ozogamicin (Mylotarg) is a conjugate of a monoclonal antibody and calicheamicin, which targets the membrane antigen CD33 in CD33-positive acute myeloid leukaemia (AML) and, after cell internalisation, releases a derivative of the cytotoxic calicheamicin component. In the US, it is approved as monotherapy in patients aged > or =60 years with a first relapse of AML who are ineligible for other cytotoxic therapy. Monotherapy with gemtuzumab ozogamicin results in complete remission (CR) or CR with incomplete platelet recovery (CRp) in approximately =25% of adults (including those aged > or =60 years) with CD33 positive AML in first relapse. Preliminary data indicate a potential role for gemtuzumab ozogamicin as a component of induction or consolidation regimens in adults and, based on an early study, in the treatment of children with AML, although randomised, controlled studies are needed. Serious adverse events, notably hepatotoxicity, characterise its tolerability profile, but gemtuzumab ozogamicin is comparatively well tolerated by most patients. Gemtuzumab ozogamicin is a valuable new treatment option for patients aged > or =60 years with CD33-positive AML in first relapse for whom other cytotoxic chemotherapy is not considered appropriate; patients with a first CR (CR1) of >12 months are likely to have the best outcome. PMID- 16266209 TI - Descriptive assessment of sleep patterns among community-living adults with mental retardation. AB - There is little information about the sleep patterns of adults who have mental retardation and are supported in the community. In the present study, direct-care staff recorded sleep behaviors of 59 adults residing in 16 suburban group homes. Based on direct observation and measurement procedures, the adults averaged 7.9 hours of sleep each evening and had low incidence of sleep problems. Sleep duration was not influenced by age, gender, degree of mental retardation, or psychiatric status. Adults taking antidepressant medication (SSRI) had fewer hours of sleep. We discuss implications of these findings and factors contributing to healthy sleep hygiene among community-living adults with mental retardation. PMID- 16266210 TI - Walking habits of adults with mental retardation. AB - The walking activity of men and women with mental retardation residing in community settings was described. Participants were 38 women (M age=.7, SD=9.5) and 65 men (M age=35.9, SD=11.2). They wore pedometers for 7 days. A 2 x 2 factorial ANOVA indicated no significant gender differences in total step counts or between participants with and those without Down syndrome. A post-hoc analysis revealed that participants walked less on Saturday than during the weekdays. Only 21.1% of the women and 21.5% of the men with mental retardation accumulated the recommended 10,000 steps per day. PMID- 16266211 TI - Model underpinning treatment for sex offenders with mild intellectual disability: current theories of sex offending. AB - Although many writers have provided a theoretical framework for treatment of mainstream sex offenders, this research has not been extended to sex offenders with mild intellectual disability. My purpose here is to bring together several research strands to provide a theoretical model for working in this field, including theories of sex offending, developmental theories for offending, and work on quality of life. In addition to dealing with issues of sex offending, researchers should also investigate developmental and societal issues crucial in the genesis of sex offending and offending in general. Because societal issues are also important for treatment, engagement and identification with society are suggested. Quality of life and issues directly related to sex offending are of central importance. PMID- 16266212 TI - Responses to Sturmey (2005) on psychotherapy evidence base for behavioral interventions: critical commentary. PMID- 16266213 TI - Psychotherapy is an essential tool in the treatment of psychiatric disorders for people with mental retardation. PMID- 16266214 TI - Proceeding with compassion while awaiting the evidence: psychotherapy and individuals with mental retardation. PMID- 16266215 TI - In support of psychotherapy for people who have mental retardation. PMID- 16266216 TI - Strange shift in the case of Daryl Atkins. PMID- 16266217 TI - Progress toward a national objective of healthy people 2010: "reduce to zero the number of children 17 years and younger living in congregate care". PMID- 16266219 TI - Promising functional readouts of immunity in a blood-stage malaria vaccine trial. PMID- 16266220 TI - Clinical value of gene NOD2/CARD15 mutations in Crohn's disease. PMID- 16266222 TI - Reliability of the Spanish version of a brief questionnaire on patient satisfaction with gastrointestinal endoscopy. AB - BACKGROUND: There has not been a validated questionnaire available in Spanish to evaluate patient satisfaction with gastrointestinal endoscopy. Our aim was to evaluate the external validity and internal consistency of the Spanish version of a questionnaire on patient satisfaction with gastrointestinal endoscopy elaborated by the American Society for Gastrointestinal Endoscopy. DESIGN: Prospective questionnaire validation study. PATIENTS AND METHODS: A total of 485 consecutive patients referred to two different hospitals for endoscopy were interviewed by telephone. Internal consistency was studied using Cronbach's alfa test and corrected item-total correlations (CITC). External validity was determined using a mailed questionnaire completed by 185 patients-correlations between telephone and postal responses were calculated, as well as the correlation with the total score obtained. RESULTS: Cronbach's alfa was 0.82 and mean CITC was 0.59. Weighted kappa values for the same questionnaire items performed by telephone or mail varied between 0.51 and 0.81. Total score correlation was 0.78. Internal consistency and external validity were not affected by differences in the administration of the questionnaire (mail or by telephone), different interviewers, type of endoscopy, or source of patients. CONCLUSIONS: The Spanish version of the ASGE questionnaire on satisfaction with endoscopy is valid, reliable, and reproducible. PMID- 16266221 TI - Association of NOD2/CARD15 mutations with previous surgical procedures in Crohn's disease. AB - OBJECTIVE: The aim of this study is to assess the importance of NOD2/CARD15 gene mutations as prognostic factors for surgical indications in Crohn's disease. PATIENTS AND EXPERIMENTAL DESIGN: A total of 165 Crohn s disease patients were studied, considering previous surgery related to Crohn's disease. We analyzed for previous surgery in global procedures as well as separately for the two main surgical indications: ileal resection and fistula treatment. The need for appendectomy was also studied. All patients were genotyped for the three CARD15 mutations, and association studies were developed using Chi-square statistics and Fisher's exact test whenever appropriate. RESULTS: Carriers of the G908R or 1007fs mutation needed surgery more frequently, both for ileal resection and fistula repair. In contrast, appendectomy was not associated with CARD15 mutations. CONCLUSIONS: As previously reported in this population, the R702W mutation does influence parameters of disease or need of surgery. The need for Crohn's disease-related surgery is higher in carriers of the G908R or 1007fs CARD15 mutation in the Galician population. Nevertheless, the frequency of these mutations does not allow their use to predict the course of disease. PMID- 16266223 TI - Total gastrectomy with or without abdominal drains. A prospective randomized trial. AB - The most common postoperative complications of total gastrectomy are esophagojejunal anastomotic leakage and subphrenic abscess. These complications are a cause of morbidity and mortality, relaparotomy, and longer postoperative stay. The use of abdominal drains is useful for the early diagnosis and management of anastomotic leaks. The aim of this study was to analyze our experience with total gastrectomy for gastric cancer in patients with and without abdominal drains, and to evaluate the results regarding postoperative morbidity, postoperative hospital stay, postoperative days for oral intake, relaparotomy and mortality. This prospective and randomized study examines the results in 60 consecutive patients (43 males and 17 females) with gastric cancer who underwent total gastrectomy in the Regional Clinical Hospital of Concepcion, Chile, between 2000 and 2003. Patients were divided into two groups: group I (without drains) and group II (two drains). We found 31 patients in group I and 29 patients in group II. The mean length of postoperative stay was 12.9 days in group I and 18.8 days in group II (p = 0.0242, s.). Morbidity was 9.7% in group I and 37.9% in group II (p = 0.0242, s.). Re-explorations were more frequent in group II (24.1%) versus group I (9.7%) (p = 0.1239, n.s.). Postoperative days for oral intake were 9.4 in group I and 12.8 in group II (p = 0.0514, n.s.) Mortality was 0% in group I and 3.4% in group II (p = 0.4833, n.s.). In our experience, morbidity and postoperative hospital stay were statistically higher in the group of patients with abdominal drains. PMID- 16266224 TI - Probiotics in arthralgia and spondyloarthropathies in patients with inflammatory bowel disease. Prospective randomized trials are necessary. AB - Arthralgias and spondyloarthropathies of the peripheral and axial joints are common in inflammatory bowel disease. Evidence for a strong association between these clinical manifestations and diseases of the joints has been provided by several clinical and epidemiological studies. Immunological studies have shown the presence of shared inflammatory cells both in the gut and the synovium in spondyloarthropathies. Genetic factors play a crucial role in the pathogenesis of spondyloarthropathies and inflammatory bowel disease. The role of the ubiquitous bacterial flora and pathogenic microorganisms present in the intestinal lumen may induce these joint diseases in patients with inflammatory bowel disease. In this review we will focus on the pathogenesis of spondyloarthropathies and arthralgia in patients suffering from inflammatory bowel disease. Based on preliminary clinical observations in patients with arthralgia and IBD, we put forward the hypothesis that probiotics may be helpful in the management of common extraintestinal manifestations such as arthralgia in patients with ulcerative colitis and Crohn's disease. PMID- 16266225 TI - IL-6 and extracellular matrix remodeling. PMID- 16266226 TI - Bruising of the esophagus as a cause of gastrointestinal bleeding in a case of heatstroke. AB - Alterations in blood clotting are a frequent complication of serious heatstroke and may result in gastrointestinal bleeding. We report the case of a 26-year-old man who was admitted to our hospital with symptoms of hyperthermia associated with encephalopathy and disseminated intravascular coagulation (DIC) after prolonged exposure to sunlight. He presented hematemesis, after which he was diagnosed with having a bruising of the esophagus that took up the upper and lower thirds, there being no other lesions in the stomach or duodenum. After supportive treatment and following the resolution of the underlying pathology, the endoscopy-revealed injuries healed with a complete normalization of the esophageal mucosa. Esophageal submucosal bruising is an exceptional cause of hematemesis in serious heatstroke not previously described in the literature. PMID- 16266227 TI - Tracheo-esophageal fistula in a patient with esophageal cancer and stent. PMID- 16266228 TI - [Spontaneous bacterial peritonitis by Listeria monocytogenes in a patient with liver cirrhosis]. PMID- 16266230 TI - [Recurrent gastric incarceration in long-term umbilical hernia]. PMID- 16266229 TI - [Fecal incontinence and female's pelvis floor: a need for a complete diagnosis]. PMID- 16266231 TI - [A new multilocular ciliated hepatic foregut cyst. Case report and review of the literature]. PMID- 16266232 TI - [Familiar penetrancy of HFE gene: four brothers of the same family affected by hereditary haemochromatosis]. PMID- 16266233 TI - Interleukins and colon cancer. PMID- 16266234 TI - Evaluation of the antitumor activity of interleukin-12 in an experimental murine model of colorectal cancer induced by 1,2 dimethyl-hydrazine (DMH). AB - OBJECTIVE: Interleukin 12 (IL-12) is a cytokine that may enhance the proliferation and cytotoxic activity of T lymphocytes and natural killer (NK) cells. A relationship between extensive intratumoral infiltration of NK cells and longer survival rates in colorectal cancer (CRC) patients was previously noted. Preliminary evidence suggests that the combined administration of IL-12 and IL-2 may produce additive immunomodulatory activity. The purpose of this study was to determine whether the systemic administration of IL-12 (+/- IL-2) may induce an immune response against CRC as induced by 1,2-dimethylhydrazine (DMH). METHODS: Sixty-five 6-week-old Wistar rats were treated with weekly subcutaneous injections of DMH for 26 weeks at a dose of 20 mg/kg of body weight. Once tumoral induction was over, the animals were randomly allocated to one of three groups: I, control; II, intraperitoneal injections of IL-12; III, intraperitoneal injections of IL-12 combined with IL-2. At 30 weeks, all surviving animals were sacrificed. We studied the following parameters in each rat--number of tumors, size of tumors, and total tumoral volume. Tumor samples were studied using the monoclonal antibody CD 57 for the detection of NK cells. The extent of NK infiltration was classified as small, less than 50 NK cells/50 high-power field (HPF); moderate, 50 to 150 NK cells/50 HPF, and extensive, more than 150 NK cells/50 HPF. RESULTS: Thirty-five rats died before completion of the carcinogen exposure, and 30 rats were randomized (10 each group). In group II, 2 animals died during treatment. All rats in groups I and III developed tumors, while in group II two rats (25%) were tumor-free. Moreover, only one rat in group II developed multiple neoplasms, in contrast with group I and group III, where six rats (60%) and seven rats (70%), respectively, had more than one tumor. We found statistically significant differences in the mean number of tumors found in group II when compared to group I (p = 0.028) and group III (p = 0.019). Other parameters measured, such as biggest tumor size and total tumoral volume were found to be lower in group II, although no statistical differences were found between groups. Only 10% of rats in group I showed moderated/extensive NK cell infiltration, vs. 60% of rats in group II (p = 0.077) and 70% in group III (p = 0.02). CONCLUSION: The administration of DMH to rodents provides a reliable and consistent means of inducing CRC that may be suitable for the evaluation of anti cancer therapies. Our findings suggest that IL-12 is effective against the development of experimental CRC. Its antineoplastic effect could be attributed to the stimulus of this cytokine on the intratumoral infiltration of NK cells. PMID- 16266235 TI - Immunopathological modifications in the rectal mucosa from an animal model of food allergy. AB - AIM: The aim is to determine immunopathological modifications in rectal mucosa from rabbit after local challenge in sensitized animals with ovalbumin (OVA). EXPERIMENTAL DESIGN: Thirty rabbits divided into three groups: G1: normal, G2: subcutaneously OVA sensitized, G3: sensitized, locally OVA challenged and sampled 4 hours after challenge. Specific anti-OVA IgE levels were evaluated by passive cutaneous anaphylaxis test (PCA). In each group 200 high microscopical power fields (HPF) were counted. Results were expressed as arithmetic mean and SE. Statistical analysis was made using Student t test. Anti-CD4, CD5, micro chain, CD25 and RLA II monoclonal antibodies were used. Avidin biotin horseradish peroxidase system was used. RESULTS: CD 4: G1: 8.3 +/- 0.06; G2: 13.4 +/- 0.08 and G3: 8.25 +/- 0.06. CD 5: G1: 7.3 +/- 0.05; G2: 9.4 +/- 0.05 and G3: 11.3 +/- 0.06. CD 25: G1: 13 +/- 0.08; G2: 15.1 +/- 0.13 and G3: 25.5 +/- 0.15. mu chain: G1: 10.4 +/- 0.06; G2: 3.8 +/- 0.02 and G3: 6.0 +/- 0.10. RLA II (DR): G1: 11.6 +/- 0.05; G2: 19.2 +/- 0.09 and G3: 19.1 +/- 0.11. In all cases, experimental groups (G2 and G3) presented statistical significant differences vs. control group (G1) (p < 0.001). CONCLUSIONS: Interleukin-2 receptor (CD25+ cells) increase in experimental groups. Cells expressing micro chain decreased in G2 and G3 likely due to activation of B cells and subsequent expression of other immunoglobulin chains in cell surface. RLA II expression is higher in G2 and G3. This receptor is considered an activation marker expressed by macrophages, T and B cells. We conclude that obtained data are important to elucidate immunopathology of local anaphylactic reaction in rectal mucosa from systemic sensitized animals. PMID- 16266236 TI - Cyclooxygenase-2 inhibition in colon experimental carcinogenesis. AB - BACKGROUND: An overexpression of cyclooxygenase-2 (COX-2) has been seen in colon tumors; therefore, COX-2 specific inhibitors may be used as preventive agents. The aim of this study was to investigate the effect of both selective and non selective COX-2 inhibitors on the incidence of colonic tumors in a model of chemical carcinogenesis in the rat. DESIGN: Experimental study with 65 male Sprague-Dawley rats randomly assigned to one of four groups: (a) control (n = 20), with chemical carcinogenesis using 1-2 dimethylhydrazine (1-2 DMH); (b) acetylsalicylic acid (ASA) (n = 15), with chemical carcinogenesis and the addition of ASA at 30 mg/kg; (c) low-dose rofecoxib (n = 15), with chemical carcinogenesis and the addition of rofecoxib at a dose of 1.2 mg/kg; (d) high dose rofecoxib (n = 15), with carcinogenesis and the addition of rofecoxib at 3 mg/kg. Carcinogenic induction was performed with 1-2 DMH at a weekly dose of 25 mg/kg for 18 weeks. The main parameter evaluated was percentage of neoplastic colonic tissue, which relates tumor surface area to colon surface area. RESULTS: Rofecoxib at a dose of 3 mg/kg significantly reduced chemical colon carcinogenesis in rats (p < 0.01). Rofecoxib in lower doses had the same effect on adenomas (p < 0.05) with no effect on adenocarcinomas. Rofecoxib reduced COX-2 expression in tumoral tissue from adenomas and adenocarcinomas (p < 0.01). CONCLUSIONS: Rofecoxib prevents chemical colon carcinogenesis in the rat, with a reduction of tumoral colonic percentage in adenocarcinomas and tumoral COX-2 expression. PMID- 16266237 TI - Laparoscopic treatment of cholelithiasis in cirrhotic patients. AB - OBJECTIVE: To assess the safety and efficacy of laparoscopy in the treatment of symptomatic cholelithiasis in patients with Child's Class A and Class B cirrhosis. STUDY DESIGN: Descriptive and retrospective study. PATIENTS: We studied 14 patients (mean age 60 yrs) with Child's Class A and Class B hepatic cirrhosis who underwent laparoscopic cholecystectomy. We analyzed the occurrence of intraoperative and postoperative complications. RESULTS: Eight patients were women (57.14%) and 6 were men (42.85%). Eight of the 14 patients presented with Child's Class B cirrhosis and 6 patients with Class A. Cholecystectomy was programmed for all patients. The average duration of surgery was 77 min. Intraoperative complications occurred in 2 patients (14.28%) in the form of liver bed bleeding. Postoperative complications were observed in 3 patients (21.42%), 2 presented with ascites which led to a worsening of Child's Class in one of them, and the third patient presented with angina-like symptoms (acute, sharp pain in the chest irradiating to the back). Mean length of hospital stay was 3 days. No postoperative morbidity or mortality occurred, and there were no conversions. CONCLUSIONS: LC (laparoscopic cholecystectomy) is a safe and effective alternative for the treatment of symptomatic cholelithiasis in patients with well compensated Child's Class A and Class B cirrhosis. Postoperative morbi-mortality is low, bleeding is unimportant, and both duration of surgical procedure and hospital stay are short. PMID- 16266238 TI - Clinical and ethical implications of genetic counselling in familial adenomatous polyposis. AB - The association of specific genetic disturbances with the development of hereditary cancer helps us to understand the risk of suffering from it, the possibility of an earlier diagnosis, and the treatment and prevention of this disease. Familial adenomatous polyposis (FAP) is a pre-neoplastic syndrome characterized by the presence of hundreds of adenomatous polyps in the colon, which develop into a carcinoma. FAP can be diagnosed using sequencing techniques to detect mutations in the germinal line of the APC (adenomatous polyposis coli) gene. The genetic diagnostic approach in families with FAP, previously followed up in the Gastrointestinal Clinic, has both advantages and disadvantages, and places us nearer the disease and patient. Disclosing the results of this genetic test entails relevant problems in clinical practice, which affect the health field and raise legal and ethical issues, along with the familial, occupational, and social implications that knowing the genetic status can have on the patient. Genetic analysis is rare in normal clinical practice, which involves errors in the interpretation of the results obtained, and during the process of genetic counselling. Specialized multidisciplinary units are necessary for the management of patients with FAP undergoing analysis and appropriate genetic counselling, thus providing an individualized service. The creation of FAP registers and protocols for this healthcare process should optimize the management of these patients and their families. PMID- 16266239 TI - Fatal portal thrombosis after laparoscopic Nissen fundoplication. AB - Portal and mesenteric vein thrombosis is a very uncommon complication of laparoscopic surgery, especially after anti-reflux procedures. We report the case of a twenty-year-old man with a history of alcohol and cocaine consumption. A Nissen fundoplication was performed. The patient received a single 20-mg dose of enoxaparin (Clexane, Aventis Pharma, Spain) two hours before surgery for antithrombotic prophylaxis. On the seventh postoperative day the patient had a portal and mesenteric venous thrombosis, which was confirmed at laparotomy, with both extensive small-intestine necrosis and partial colon necrosis. Despite anticoagulant therapy, the patient died 24 hours later. Surgical findings were confirmed at necropsy. Portal and mesenteric venous thrombosis is an uncommon but severe and even fatal complication after laparoscopic anti-reflux surgery. When other pro-thrombotic, predisposing conditions such as laparoscopic surgery and cocaine consumption are present, the usual prophylactic doses of low molecular weight heparin might not be sufficient to protect against this life-threatening complication. PMID- 16266240 TI - Severe intestinal involvement in Wegener's granulomatosis with negative c-ANCAs. PMID- 16266241 TI - [Hydatidic cyst fistulized to colon]. PMID- 16266242 TI - [Biliary fistula associated with hepatorrhaphy for liver injury. A possible postural management]. PMID- 16266243 TI - [Gastrointestinal stromal tumor of the gallbladder]. PMID- 16266244 TI - [Chronic autoimmune pancreatitis: an entity of difficult differential diagnosis]. PMID- 16266245 TI - [Primary biliary cirrhosis, "sicca" sindrome and autoimmune hemolytic anemia]. PMID- 16266246 TI - [Primary sclerosing cholangitis as a cause of bile duct confluence pseudotumor]. PMID- 16266247 TI - Selective effect of inhibition of literal or numerical irrelevant information in children with attention deficit hyperactivity disorder (ADHD) or arithmetic learning disorder (ALD). AB - This study investigated reasons why children with attention deficit hyperactivity disorder (ADHD) or arithmetic learning disorder (ALD) have difficulties in solving arithmetic word problems. In particular, the aim of this study was to verify whether these difficulties are due to a working memory deficit and defective inhibition of irrelevant information included in the problem wording. Furthermore, the study was geared to test whether children with ADHD or ALD have a specific disability in recalling and handling numerical or literal information. In an attempt to provide an answer to these questions, three groups of children were tested: children with ADHD, children with ALD, and a group of children achieving at normal levels. They were presented with a battery of arithmetic word problems containing irrelevant information (using either numerical or literal information). Children were asked to recall relevant information within the texts and then solve the problems. Children with ADHD recalled significantly more irrelevant literal information. Both children with ADHD and ALD recalled significantly more irrelevant numerical information. On the other hand, in the phase of problem solving, children with ADHD were more impaired by irrelevant literal information (which overloads memory), whereas children with ALD were more impaired by irrelevant numerical information (which may elicit the execution of wrong arithmetic procedures). PMID- 16266248 TI - Assessing executive dysfunction in girls with fragile X or Turner syndrome using the Contingency Naming Test (CNT). AB - The aim of this study was to examine executive function (EF) skills in girls with fragile X or Turner syndrome, using the Contingency Naming Test (CNT). The CNT is a Stroop-like task involving a 1- or 2-attribute contingency rule. We predicted that girls with fragile X would make errors reflecting poor cognitive flexibility and working memory limitations. We predicted that girls with Turner syndrome would have sufficient cognitive flexibility to perform the CNT accurately, but would have difficulty with verbal inhibition and would thus make more self corrections than girls in a comparison group. The hypotheses were partially supported: relative to their Full Scale IQ-matched comparison group, girls with fragile X or Turner syndrome were slower on the warm-up naming task; girls with fragile X made more errors on the 1-attribute task, and girls with Turner syndrome were less efficient on both the 1- and 2-attribute tasks, without making more self-corrections. These results support previous findings of executive dysfunction associated with fragile X or Turner syndrome. The results suggest that both low IQ and fragile X status contribute to working memory limitations in girls with fragile X and that EF inefficiency in girls with Turner syndrome is due to both working memory limitations and slower response times. PMID- 16266249 TI - Implicit memory development in school-aged children with attention deficit hyperactivity disorder (ADHD): conceptual priming deficit? AB - Previous research has shown that children with attention deficit hyperactivity disorder (ADHD) often demonstrate performance deficits on effortful, strategic memory tasks, whereas relatively rote tasks of memory reveal no such deficit. Thus far, research in this domain has focused primarily on explicit memory. This study examined performance on multiple measures of implicit and explicit memory in children aged 7 to 14 years with and without ADHD. Memory for words and pictures was assessed at 15-min and 24-hr intervals. ADHD and non-ADHD groups performed similarly on tests of explicit memory (category-cued recall and recognition) and on perceptual aspects of implicit memory (word stem completion and picture fragment identification) as a function of age, retention interval, and stimulus format (i.e., picture or word). However, there was no evidence of priming on a conceptual implicit memory test (category exemplar generation) for boys with ADHD. This type of conceptual task, which is likely mediated by frontal systems, may indicate a unique memory deficit associated with ADHD. PMID- 16266250 TI - Disorganization: the forgotten executive dysfunction in high-functioning autism (HFA) spectrum disorders. AB - Executive function (EF) abilities were investigated in 72 children with high functioning autism (HFA) spectrum disorders through the collection of parent ratings and performance on laboratory measures of EF. In addition, discrepancy analysis was used to isolate executive functioning on tasks that carry multiple demands. Comparison of HFA and Asperger Disorder (AD) groups did not reveal consistent differences in EF. Results did indicate global EF deficits in the combined group of children with HFA and AD. Within the EF domain, specific deficits in flexibility and organization were most prominent. PMID- 16266251 TI - Response inhibition after traumatic brain injury (TBI) in children: impairment and recovery. AB - Children who experience traumatic brain injury (TBI) often show cognitive impairments postinjury, some of which recover over time. We examined the recovery of motor response inhibition immediately following TBI and over 2 years. We assessed the role of injury severity, age at injury, and lesion characteristics on initial impairment and recovery while considering the role of pre-injury psychiatric disorder. Participants were 136 children with TBI aged 5-16 years. Latency of motor response inhibition was measured with the stop-signal task within 1 month of the injury and again at 3, 6, 12, and 24 months. The performance of the TBI participants at each measurement occasion was standardized with 117 children of similar age, but without injury. Residualized latency scores were calculated. Growth curve analyses showed an initial impairment in response inhibition and improvement over the 2 years following injury. Younger TBI patients were initially more impaired although they exhibited greater recovery of response inhibition than did older TBI patients. Longer duration of coma, but not reactivity of pupils or Glasgow Coma Scale score, predicted initial deficit. Lesion characteristics or pre-injury attention deficit hyperactivity disorder did not predict initial impairment or recovery. Replication with longitudinal testing of a comparison group of children sustaining extracranial injury is necessary to confirm our findings. PMID- 16266252 TI - Sex differences in cognitive functioning in velocardiofacial syndrome (VCFS). AB - Sex differences in cognitive functioning were investigated in children with velocardiofacial syndrome (VCFS), a genetic defect caused by a microdeletion on chromosome 22q.11. The study population consisted of six groups: 50 boys with VCFS (M = 11.1, SD = 2.7), 40 girls with VCFS (M = 10.8, SD = 2.5), 13 male siblings of the participant with VCFS (M = 12.3, SD = 1.9), 17 female siblings of the participant with VCFS (M = 12.2, SD = 1.9), and a race- and gender-ratio matched sample of 28 boy community control participants (M = 10.7, SD =2.4) and 19 girl community control participants (M = 9.2, SD =2.3). Each participant received a psychological assessment including intellectual and academic achievement as well as structural magnetic resonance imaging of his or her brain. Our results indicate that boys with VCFS may be more cognitively affected than girls. In addition, and although cross-sectional in nature, our results document a negative association between age and cognitive functioning in girls with VCFS but not in boys. Sex differences in frontal lobe volume are generally seen in the general population between boys and girls (boys > girls) and across all three samples, this trend emerged. Relative to boys with VCFS, girls with VCFS may be less cognitively affected, although age is negatively associated with cognitive functioning in girls with VCFS but not boys, suggesting that girls with VCFS may fail to maintain this cognitive advantage over boys. PMID- 16266253 TI - Error-related negativity in children: effect of an observer. AB - This research assessed the effect of an observer upon error-related negativity (ERN) in 20 children (ages 7-11): 9 worked alone and 11 worked under the observation of a friend of theirs (alone/audience groups). Erroneous motor responses were recorded to Go and No-Go visual stimuli (triangles in different orientations) at three brain sites (Fz, Cz, and Pz). Results for children were consistent with past adult studies in showing a significantly larger ERN only to erroneous responses in both the alone and the audience groups. Children in the audience group produced larger ERNs than those in the alone group. Furthermore, older children (ages 9-11) produced larger ERNs than younger children (ages 7-8) in both groups. These findings were discussed in the context of an action monitoring system that regulates responses, detects errors, and entrains affective responses associated with correct and incorrect responses. PMID- 16266266 TI - Programmed cell death in plants: ultrastructural changes in pea guard cells. AB - Treatment with cyanide of epidermal peels isolated from pea leaves resulted in destruction of nuclei in the guard cells of stomata, which is visible with a light microscope. The process was accelerated by illumination. Electron microscopy revealed significant CN--induced changes in the ultrastructure of guard cells, which increased with time. Margination of chromatin, which is one of the first signs of apoptosis, was observed in the guard cells even after 1 h incubation of the isolated epidermis with CN-. Subsequent chromatin condensation, swelling of the endoplasmic reticulum with formation of large tanks covered with ribosomes, changes in the structure of dictyosomes, and a slight swelling of mitochondria were observed after 3 h of the epidermis incubation with CN-. After 6 h of incubation with CN-, the bulk volume of the guard cells was filled with vacuoles, the cytoplasm occupied the thin marginal layer, the nucleus was in the center similarly to the control experiment, but it was polylobal, extended in narrow cytoplasmic bands, and, despite the loss of the nuclear envelope integrity, appeared to be a self-dependent structure. In the envelope-free open regions of the nucleus, mitochondria and chloroplasts directly contacted with chromatin. Much like the cell nucleus, chloroplasts lost the integrity of the membrane, but did not swell and retained the stroma and integrity of the thylakoid system. An antioxidant di-tert-butyl-4-hydroxytoluene prevented ultrastructural changes in the cells observed after 6 h of incubation with CN-. Thus, the CN--induced death of the guard cells of stomata occurs through the mechanism of apoptosis. PMID- 16266265 TI - The p53 gene family: control of cell proliferation and developmental programs. AB - For a quarter of a century the gene p53 has attracted close attention of scientists who deal with problems of carcinogenesis and maintenance of genetic stability. Multicellular organisms on our planet owe their rich evolution in many respects to the ability of this gene to protect cells from oncogenic transformation and harmful changes in DNA. A relatively recent discovery of structural p53 homologs, the genes p63 and p73, which seem to have more ancient roots, has roused keen interest in their function. Do they carry out oncosuppressor functions in partnership with p53 or do they possess their own specific functions? This review analyzes data on p53, p63, and p73 functional activity at the levels of the molecule, cell, and whole organism with the accent on examination of specific p63/p73 targets indicating a unique role of these genes in control of developmental processes. PMID- 16266267 TI - Accumulation of polyphosphates and expression of high molecular weight exopolyphosphatase in the yeast Saccharomyces cerevisiae. AB - The effect of cultivation time and concentration of inorganic phosphate (P(i)) in the culture medium on the accumulation of polyphosphates (polyP) and the activity of two cytosolic exopolyphosphatases of the yeast Saccharomyces cerevisiae was studied: an exopolyphosphatase of 40 kD encoded by PPX1 and a high molecular weight exopolyphosphatase encoded by another gene. Depletion of polyP in the cells on P(i) starvation is a signal factor for the accumulation of polyP after the subsequent addition of 5-20 mM P(i) and glucose to the cells or spheroplasts. A high activity of both exopolyphosphatases does not prevent the accumulation of polyP. The expression of the high molecular weight exopolyphosphatase is due to the acceleration of metabolism in cells that have reached the stage of growth deceleration on the addition of P(i) and glucose or complete culture medium. This process may occur independently from the accumulation of polyP. The activity of exopolyphosphatase PPX1 depends less on the mentioned factors, decreasing 10-fold only under conditions of phosphate surplus at the stationary growth stage. PMID- 16266268 TI - Uncoupler of oxidative phosphorylation prolongs the lifespan of Drosophila. AB - The effect of a moderate ("soft") uncoupling of mitochondria on the lifespan and some parameters of biological age of Drosophila melanogaster strain Oregon was studied. Addition of the uncoupler 2,4-dinitrophenol (DNP) to the nutritional mixture of larvae significantly increased the average lifespan of the flies without changing their maximal lifespan. DNP significantly increased the rate of oxygen consumption by isolated mitochondria and tissue homogenates of the flies in state 4 (of Chance). DNP also decreased the activity of alcohol dehydrogenase (a parameter of flies' biological age) in the tissue homogenates, especially on octanol as the reaction substrate. However, being deprived of food the DNP treated flies displayed a markedly decreased viability as compared to the control flies. On the whole, the results suggest that "soft" uncoupling of mitochondria may increase the lifespan. PMID- 16266269 TI - Metabolic activity of macrophages infected with hantavirus, an agent of hemorrhagic fever with renal syndrome. AB - Monocytes/macrophages are thought to play an important role in pathogenesis of viral infections. These cells are involved in distribution and persistence of viruses in the organism and also influence the regulation of immune reactions. The functional and enzymatic activities of macrophages infected with an agent of hemorrhagic fever with renal syndrome were analyzed for the first time. This disease is caused by a virus of the Hantavirus genus, the Bunyaviridae family. Activities of ectoenzymes 5 -nucleotidase and ATPase of the plasma membrane of the hantavirus-infected macrophages decreased along with the antigen accumulation in the infected cells. The contact of phagocytes with hantavirus resulted in activation in the cells of the oxygen-dependent metabolism and NO-synthase. The NO-synthase-dependent system of the infected macrophages was activated earlier than their oxygen-dependent system. The intracellular contents of acid and alkaline phosphatases increased within the first hours after the infection. The bactericidal activity of the hantavirus-infected macrophages relatively to Staphylococcus aureus increased during the specific antigen accumulation in the phagocytes. Thus, the infection of macrophages with hantavirus was associated with intracellular metabolic changes. PMID- 16266270 TI - Generation of free radicals during decomposition of hydroperoxide in the presence of myeloperoxidase or activated neutrophils. AB - It was shown with the spin trap alpha-(4-pyridyl-1-oxide)-N-tert-butylnitrone that myeloperoxidase (MPO) in the presence of its substrates H2O2 and Cl- as well as activated neutrophils destroy tert-butyl hydroperoxide producing two adducts of O-centered radicals which were identified as peroxyl and alcoxyl radicals. Inhibitory analysis performed with traps of hypochlorite (taurine and methionine), free radical scavengers (2,6-di-tret-butyl-4-methylphenol and mannitol), and MPO inhibitors (salicylhydroxamic acid and 4-aminobenzoic acid hydrazide) revealed that the destruction of the hydroperoxide group in the presence of isolated MPO or activated neutrophils was directly caused by the activity of MPO: some radical intermediates appeared as a result of the chlorination cycle of MPO at the stage of hypochlorite generation, whereas the other radicals were produced independently of hypochlorite, presumably with involvement of the peroxidase cycle of MPO. The data suggest that the activated neutrophils located in the inflammatory foci and secreting MPO into the extracellular space can convert hydroperoxides into free radicals initiating lipid peroxidation and other free radical reactions and, thus, promoting destruction of protein-lipid complexes (biological membranes, blood lipoproteins, etc.). PMID- 16266272 TI - Hemolysis of human red blood cells by riboflavin-Cu(II) system: enhancement by azide. AB - Photoactivated riboflavin in the presence of Cu(II) generates reactive oxygen species (ROS) which can hemolyze human red blood cells (RBC). In the present work we examined the effect of sodium azide (NaN3) on RBC in the presence of riboflavin and Cu(II). The addition of NaN3 to the riboflavin-Cu(II) system enhanced K+ loss and hemolysis. The extent of K+ loss and hemolysis were time and concentration dependent. Bathocuproine, a Cu(I)-sequestering agent, inhibited the hemolysis completely. Among various free radical scavengers used to identify the major ROS involved in the reaction, thiourea was found to be the most effective scavenger. Thiourea caused almost 85% inhibition of hemolysis suggesting that *OH is the major ROS involved in the reaction. Using spectral studies and other observations, we propose that when NaN3 is added to the riboflavin-Cu(II) system, it inhibits the photodegradation of riboflavin resulting in increased *OH generation. Also, the possibility of azide radical formation and its involvement in the reaction could not be ruled out. PMID- 16266271 TI - Cloning and expression of small cDNA fragment encoding strong antiviral peptide from Celosia cristata in Escherichia coli. AB - A small cDNA fragment containing a ribosome-inactivating site was isolated from the leaf cDNA population of Celosia cristata by polymerase chain reaction (PCR). PCR was conducted linearly using a degenerate primer designed from the partially conserved peptide of ribosome-inactivating/antiviral proteins. Sequence analysis showed that it is 150 bp in length. The cDNA fragment was then cloned in a bacterial expression vector and expressed in Escherichia coli as a ~57 kD fused protein, and its presence was further confirmed by Western blot analysis. The recombinant protein was purified by affinity chromatography. The purified product showed strong antiviral activity towards tobacco mosaic virus on host plant leaves, Nicotiana glutinosa, indicating the presence of a putative antiviral determinant in the isolated cDNA product. It is speculated that antiviral site is at, or is separate but very close to, the ribosome-inactivating site. We nominate this short cDNA fragment reported here as a good candidate to investigate further the location of the antiviral determinants. The isolated cDNA sequence was submitted to EMBL databases under accession number of AJ535714. PMID- 16266273 TI - Effect of lactoferrin on the ferroxidase activity of ceruloplasmin. AB - The effects of various forms of lactoferrin (Lf) interacting with ceruloplasmin (Cp, ferro-O2-oxidoreductase, EC 1.16.3.1) on oxidase activity of the latter were studied. Comparing the incorporation of Fe3+ oxidized by Cp into Lf and serum transferrin (Tf) showed that at pH 5.5 apo-Lf binds the oxidized iron seven times and at pH 7.4 four times faster than apo-Tf under the same conditions. Apo-Lf increased the oxidation rate of Fe2+ by Cp 1.25 times when Cp/Lf ratio was 1 : 1. Lf saturated with Fe3+ or Cu2+ increased the oxidation rate of iron 1.6 and 2 times when Cp to holo-Lf ratios were 1 : 1 and 1 : 2, respectively. Upon adding to Cp the excess amounts of apo-Lf (Cp/apo-Lf < 1 : 1) or of holo-Lf (Cp/holo-Lf < 1 : 2) the oxidation rate of iron no longer changed. Complex Cp-Lf demonstrating ferroxidase activity was discovered in breast milk. PMID- 16266275 TI - Malate dehydrogenase from the thermophilic bacterium Vulcanithermus medioatlanticus. AB - Thermostable dimeric malate dehydrogenase (MDH) was isolated from the microorganism of hydrothermal vents Vulcanithermus medioatlanticus. The enzyme was electrophoretically homogeneous and possessed the specific activity of 6.9 U/mg. The large molecular weight of the subunits (55 kD) is likely to provide the rigidity of the enzyme structure (the activation energy of the enzymatic reaction is 32.6 kJ/mol). The thermophilic MDH differs little from the mesophilic enzyme in terms of kinetic and regulatory characteristics. PMID- 16266274 TI - Influence of osmolytes on inactivation and aggregation of muscle glycogen phosphorylase b by guanidine hydrochloride. Stimulation of protein aggregation under crowding conditions. AB - The effects of the osmolytes trimethylamine-N-oxide (TMAO), betaine, proline, and glycine on the kinetics of inactivation and aggregation of rabbit skeletal muscle glycogen phosphorylase b by guanidine hydrochloride (GuHCl) have been studied. It is shown that the osmolytes TMAO and betaine exhibit the highest protective efficacy against phosphorylase b inactivation. A test system for studying the effects of macromolecular crowding induced by osmolytes on aggregation of proteins is proposed. TMAO and glycine increase the rate of phosphorylase b aggregation induced by GuHCl. PMID- 16266276 TI - Isolation and characterization of a new extracellular bacteriolytic endopeptidase of Lysobacter sp. XL1. AB - The previously unstudied bacteriolytic enzyme L(4) was isolated from the culture liquid of the bacterium Lysobacter sp. XL1 in electrophoretically homogeneous state. The enzyme L(4) is a diaminopimelinoyl-alanine endopeptidase relative to peptidoglycan of Lysobacter sp. XL1. The enzyme is an alkaline protein of approximately 21 kD. The N-terminal amino acid sequence of the enzyme has been determined - A V V N G V N Y V Gx T T A ... The maximal activity of the enzyme was observed in 0.05 M Tris-HCl at pH 8.0 and 50-55 degrees C. The half inactivation temperature of the enzyme is 52 degrees C. The endopeptidase L(4) is not a metalloenzyme since it is not affected by EDTA. The enzyme is inhibited by p-chloromercuribenzoic acid by 72% and by phenylmethylsulfonyl fluoride by 43%, which indicates the involvement of serine and thiol groups in its functioning. PMID- 16266277 TI - Polyclonal antibodies against a structure mimicking the covalent linkage unit between picornavirus RNA and VPg: an immunochemical study. AB - We propose that therapy of patients with anticancer drugs that poison DNA topoisomerases induces formation of covalent complexes of cellular RNAs and DNA topoisomerases. The appearance of these complexes can be detected with antibodies against a synthetic hapten mimicking the covalent linkage unit Tyr-pU(p) of picornavirus RNA and VPg. We synthesized hapten [N(Ac),CO(NH2)]Tyr-(5 P --> O)Up O-(CH2)6NH2, conjugated it with BSA, and immunized rabbits with the antigen obtained. The raised polyclonal antibodies were purified by successive affinity chromatography on BSA-Sepharose and hapten-Sepharose columns. Target antibodies recognized hapten and encephalomyocarditis virus RNA-VPg complex specifically as found using the dot-immunogold method. We believe that these antibodies might be useful to study mechanism of picorna and similar virus RNA synthesis. The discovery and qualitative determination of the cellular RNA-DNA topoisomerases covalent complexes with these antibodies might be useful to monitor therapy efficacy by drugs "freezing" dead-end complexes of DNA topoisomerases and nucleic acids and to understand the mechanism of DNA topoisomerase poisoning in situ. PMID- 16266278 TI - Anionic polymers of the cell wall of Brevibacterium linens VKM Ac-2159. AB - Unsubstituted 1,3-poly(glycerol phosphate) and two sugar-1-phosphate polymers were identified in the cell wall of Brevibacterium linens VKM Ac-2159 by NMR spectroscopy and chemical methods. A monomer of one of the sugar-1-phosphate polymers has the branched repeating unit of the following structure: -4)-[beta-D GlcpNAc-(1-->3)]-alpha-D-Glcp-(1-P-. The repeating unit of another sugar-1 phosphate polymer has a linear structure consisting of alternating beta- and alpha-N-acetylglucosamine residues: -4)-beta-D-GlcpNAc-(1-->6)-alpha-D-GlcpNAc-(1 P-. Some part of the beta-N-acetylglucosaminyl residues bear O-ester-bound succinic acid residues at C-3. The identified sugar-1-phosphate polymers have not been described earlier in cell walls of other bacteria. PMID- 16266279 TI - Cancer as a programmed death of an organism. AB - The hypothesis introduces the idea that there is a critical level of mutagenesis that triggers a program of organism death by means of proliferation of killer cells. Similarly to apoptosis, which is an altruistic suicidal act of a faulty cell threatening the stability of a multicellular organism, a malignant tumor is an altruistic suicide of an individual carrier of harmful alleles threatening genetic stability of the population. PMID- 16266280 TI - Transcription factor GATA-3 is involved in repression of promoter activity of the human interleukin-4 gene. AB - GATA-3 was shown to bind to two sites of the IL-4 gene promoter in human T-cell lines PER-117 and Jurkat. A motif located in the region of position -860 and responsible for GATA-3 binding was detected for the first time. Mutation or deletion of this site increased the promoter activity. The findings suggest a direct involvement of GATA-3 in regulation of the human IL-4 gene transcription as a repressor of the promoter activity. PMID- 16266283 TI - CREB, synapses and memory disorders: past progress and future challenges. AB - In neurons, appropriate long-term adaptive responses to changes in the environment require the conversion of extracellular stimuli into discrete intracellular signals. Many of these signals involve the regulation of gene expression. The cAMP responsive element binding protein (CREB) is a nuclear transcription factor that modulates transcription of genes containing cAMP responsive elements (CRE sites) in their promoters. CREB is a key part of many intracellular signaling events that critically regulate many neural functions. Numerous studies on invertebrates and vertebrates demonstrate that CREB is critical for long-term memory. Here, we review the key features of CREB-dependent transcription and critically evaluate the data examining the roles of CREB in different forms of plasticity, including long-term memory in mammals. Because learning and memory have been linked to specific types of synaptic plasticity in several species, we also review studies on the role of CREB in long-term facilitation in Aplysia and in hippocampal long-term potentiation (LTP). Several human cognitive disorders have been linked to alterations of CREB-regulated gene expression. Therefore, we explore the possibility of targeting CREB function in developing novel treatment strategies. Finally, we highlight areas of research on CREB that are ripe for further advancement. PMID- 16266282 TI - Low density lipoprotein receptor-related proteins (LRPs), Alzheimer's and cognition. AB - This review will focus primarily on the role of the low density lipoprotein receptor-related protein (LRP-1) in neuronal synapse formation and function in Alzheimer's Disease (AD). We review the role that its ligands may have in cognition or AD: apolipoprotein E (ApoE), alpha2-macroglobulin, Transforming Growth Factor-Beta (TGFbeta, Tissue Plasminogen Activator (tPA), insulin growth factor binding protein-3 (IGFBP-3), which all bind LRP-1 and apolipoprotein J (ApoJ), which is a ligand for LRP-2. After reviewing its role as a signaling receptor, we discuss the connection between LRP and the NMDA glutamate receptor via the post synaptic density 95 (PSD-95) neuronal scaffold protein and the implications it may have for memory and cognition. Finally, we discuss the evidence supporting a role for LRP in AD. Although the evidence for LRP as a genetic risk factor is weak, many of its ligands impose genetic risk, and have been implicated in AD pathogenic cascades. We discuss the role of LRP in amyloid precursor protein (APP) processing and production of beta-amyloid (Abeta. We identify LRP ligands that accelerate aggregation of toxic Abeta species. LRP mediates crucial pathways in AD pathogenesis such as Abeta clearance, Abeta uptake, intraneuronal Abeta accumulation and Abeta-associated neuron death. Interestingly, the TGFbeta -V receptor is LRP-1. Data show that one critical ligand TGFbeta2, associated with neurodegeneration in amyloid diseases, induces LRP expression in PC12 cells. Data from rodent infusion models demonstrate the impact of TGFbeta2 in modifying Abeta- induced Long Term Potentiation (LTP) responses, presynaptic proteins, lipid peroxidation, gliosis and staining for neuronal nuclei. The evidence supports a complex and significant role of LRP in cognition and AD. PMID- 16266284 TI - Memantine: a therapeutic approach in treating Alzheimer's and vascular dementia. AB - Memantine has been clinically used in the treatment of organic disorders in Germany for over ten years and has now been approved in Europe and also in the US for moderate to severe Alzheimer's disease. The rationale for this indication is strongly related to the physiological and pathological role of glutamate in neurotransmission. Glutamate is an agonist of NMDA, kainate and AMPA (ionotropic) receptors, where its influence on NMDA receptors plays an important role with regard to neuronal plasticity effecting memory and learning. Excessive levels of glutamate result in neurotoxicity, in part by overactivation of NMDA receptors. Memantine acts as an uncompetitive antagonist of NMDA receptors and therefore compensates for this overactivation. Furthermore, memantine is a neuroprotective agent in various animal models based on both neurodegenerative and vascular processes, as it ameliorates cognitive and memory deficits. Memantine was effective and safe in several clinical studies, particularly in Alzheimer's disease. The compound is completely absorbed after oral intake and undergoes little metabolism. Having a low probability for drug-drug interactions, memantine, in principle, is suited for elderly patients exposed to multiple therapeutic therapies. PMID- 16266285 TI - Cannabinoids. AB - Since the discovery of an endogenous cannabinoid system, research into the pharmacology and therapeutic potential of cannabinoids has steadily increased. Two subtypes of G-protein coupled cannabinoid receptors, CB(1) and CB(1), have been cloned and several putative endogenous ligands (endocannabinoids) have been detected during the past 15 years. The main endocannabinoids are arachidonoyl ethanolamide (anandamide) and 2-arachidonoyl glycerol (2-AG), derivatives of arachidonic acid, that are produced "on demand" by cleavage of membrane lipid precursors. Besides phytocannabinoids of the cannabis plant, modulators of the cannabinoid system comprise synthetic agonists and antagonists at the CB receptors and inhibitors of endocannabinoid degradation. Cannabinoid receptors are distributed in the central nervous system and many peripheral tissues, including immune system, reproductive and gastrointestinal tracts, sympathetic ganglia, endocrine glands, arteries, lung and heart. There is evidence for some non-receptor dependent mechanisms of cannabinoids and for endocannabinoid effects mediated by vanilloid receptors. Properties of CB receptor agonists that are of therapeutic interest include analgesia, muscle relaxation, immunosuppression, anti-inflammation, antiallergic effects, improvement of mood, stimulation of appetite, antiemesis, lowering of intraocular pressure, bronchodilation, neuroprotection and antineoplastic effects. The current main focus of clinical research is their efficacy in chronic pain and neurological disorders. CB receptor antagonists are under investigation for medical use in obesity and nicotine addiction. Additional potential was proposed for the treatment of alcohol and heroine dependency, schizophrenia, conditions with lowered blood pressure, Parkinson's disease and memory impairment in Alzheimer's disease. PMID- 16266286 TI - Increasing testosterone levels and effects on cognitive functions in elderly men and women: a review. AB - Low testosterone (T) levels may predispose to Alzheimer disease (AD), but it is unclear whether this is a co-morbid effect due to cachexia, subclinical hyperthyroidism or other co-morbidity. The biological plausibility for potential protective effects of T on brain functions is substantial. In addition, higher levels of gonadotropins found in older cases with AD suggest that low levels of T are not due to brain degeneration and that the hypothalamic-pituitary-gonadal (HPG) axis is still intact. Men genetically at risk for AD were also already found to have lower levels of T. However, despite having lower levels of T, women do not show accelerated cognitive decline with age when compared to men. In addition, castration has not necessarily shown a decline in cognitive functions; some studies even found improvement of memory recall. Age may be an important factor when assessing optimal levels of T and several studies suggest that free or bioavailable T may be a better marker than total T levels when investigating associations of androgen activity with cognitive function. Small-scale T intervention trials in elderly men with and without dementia suggest that some cognitive deficits may be reversed, at least in part, by short term T supplementation. Age and prior hypogonadism may play an important role in therapy success and these factors should be investigated in more detail in future large scale randomized controlled studies. For elderly women, T treatment does not seem to have additional benefits over estrogen treatment for postmenopausal complaints and cognitive decline and may increase cardiovascular disease. PMID- 16266287 TI - Protein kinase C isozymes: memory therapeutic potential. AB - PKC plays an important role in many types of learning and memory. Evidence has been provided that PKC activation and translocation are induced in associative learning tasks. PKC inhibition, on the other hand, impairs learning and memory, consistent with the observations that transgenic animal models with a particular PKC isoform deficit exhibit impaired capacity in cognition. The dramatic impact of PKC pharmacology on learning and memory is further emphasized by a regulatory role of PKC isozymes in amyloid production and accumulation. Recent study reveals that PKC activation greatly reduces neurotoxic amyloid production and accumulation. PKC activators, therefore, may have important therapeutic values in the treatment of dementia, especially when fine-tuning of selective isoform activity can be effectively achieved pharmacologically, with further development of isozymes-specific agents. The success of antidementia therapy with agents that act on PKC signaling cascades depends on whether such agents at their effective doses would significantly disrupt or interfere with other vital functions that rely on a narrow range of PKC activities. PMID- 16266288 TI - Cholesterol and apoe: a target for Alzheimer's disease therapeutics. AB - Alzheimer's disease (AD) is a debilitating disease that affects many people. In order to reduce the number of people diagnosed with this disease, drug strategies need to be implemented that target early steps in disease pathogenesis. Elevated cholesterol levels and presence of the apolipoprotein E eta4 allele increase AD risk. How these two factors may contribute to AD pathogenesis and some therapeutic strategies for alleviating AD risk will be discussed. PMID- 16266289 TI - Abeta immunotherapy and other means to remove amyloid. AB - The amyloid cascade hypothesis postulates that accumulation of beta-amyloid (Abeta) plays a key role in the development of Alzheimer's disease (AD). Accordingly, much effort has gone into reducing the amyloid burden, especially in transgenic mice expressing mutations in human amyloid precursor protein. Such mice develop amyloid plaques but not neurofibrillary tangles. Immunization with Abeta and other inflammatory stimuli, inhibitors of Abeta formation, cholesterol lowering agents, beta-sheet breaker peptides, antioxidants and various miscellaneous agents have been found to reduce the more soluble Abeta in such transgenic mice. Whether they would affect the more consolidated, cross-linked Abeta of AD and, if they did, whether that would really prove an effective treatment for the disease remains for future research to determine. PMID- 16266290 TI - Neuroactive flavonoids interacting with GABAA receptor complex. AB - Classical benzodiazepines (BZs) are the most widely prescribed drugs acting on the central nervous system (CNS). They exert their therapeutic effects via binding to the BZ-site of GABAA receptors, and allosterically modulating the chloride flux through the ion channel complex. Given the multiple actions of classical BZs, the serious limitations to their usefulness have directed much research into development of novel ligands for the BZ-site with retained therapeutic effectiveness and minimal side effects. From the studies of CNS active chemical constituents of medicinal herbs, some members of the family of flavonoids were demonstrated to have moderate binding affinities for the BZ-site. In vivo studies revealed that these compounds were mostly partial agonists of GABAA receptors, and only a few flavonoids were shown to possess antagonistic activities. At effective anxiolytic doses, the actions of partial agonistic flavonoids were often not accompanied by sedative and myorelaxant side effects. Based on structure-activity relationship (SAR) studies, incorporation of electronegative groups to the C6 and C3' on the flavone backbone was found to yield significant increases in the binding affinities for the BZ-site. It was also shown that 2'-hydroxyl was a critical moiety on flavonoids with regard to BZ site binding. These have guided the identification of several synthetic flavonoids with high BZ-site binding affinity and in vivo activity, and further quantitative SAR studies resulted in the development of several pharmacophore models. This review attempts to summarize these findings, which has led to the establishment of flavonoids as potential therapeutics for GABAA receptor-mediated disorders. PMID- 16266291 TI - Small transthyretin (TTR) ligands as possible therapeutic agents in TTR amyloidoses. AB - In transthyretin (TTR) amyloidosis TTR variants deposit as amyloid fibrils giving origin, in most cases, to peripheral polyneuropathy, cardiomyopathy, carpal tunnel syndrome and/or amyloid deposition in the eye. More than eighty TTR variants are known, most of them being pathogenic. The mechanism of TTR fibril formation is still not completely elucidated. However it is widely accepted that the amino acid substitutions in the TTR variants contribute to a destabilizing effect on the TTR tetramer molecule, which in particular conditions dissociate into non native monomeric intermediates that aggregate and polymerize in amyloid fibrils that further elongate. Since this is a multi-step process there is the possibility to impair TTR amyloid fibril formation at different stages of the process namely by tetramer stabilization, inhibition of fibril formation or fibril disruption. Till now the only efficient therapy available is liver transplant when performed in an early phase of the onset of the disease symptoms. Since this is a very invasive therapy alternatives are desirable. In that sense, several compounds have been proposed to impair amyloid formation or disruption. Based on the proposed mechanism for TTR amyloid fibril formation we discuss the action of some of the proposed TTR stabilizers such as derivatives of some NSAIDs (diflunisal, diclofenac, flufenamic acid, and derivatives) and the action of amyloid disrupters such as 4'-iodo-4'-deoxydoxorubicin (I-DOX) and tetracyclines. Among all these compounds, TTR stabilizers seem to be the most interesting since they would impair very early the process of amyloid formation and could also have a prophylactic effect. PMID- 16266292 TI - Neuronal plasticity, stress and depression: involvement of the cytoskeletal microtubular system? AB - In susceptible individuals, stressors can increase the risk of onset of depression and recent brain imaging studies have shown morphometric alterations in the limbic system of patients affected by depression. The volume loss observed in the hippocampus of depressed individuals suggests a possible involvement of structural neuronal plasticity in the pathogenesis of depression. Stressful conditions in animals can result in impaired structural neuronal plasticity in the hippocampus, characterised by retraction of apical dendrites and decreased neurogenesis. The intrinsic dynamic instability of the cytoskeletal microtubular system is essential for neuronal remodelling and plasticity. We have recently shown that both acute and chronic stress decrease microtubular dynamics in the rat hippocampus. Other authors have demonstrated that proteins functionally involved in the regulation of microtubule dynamics can be altered by stress in the rodent hippocampus. Furthermore, the existence of a link between stress induced microtubular changes and depression is further strengthened by evidence showing that both acute and chronic treatment with antidepressant drugs can affect the expression of microtubular proteins. The present review will introduce a growing body of evidence suggesting that stress-induced alterations in neuronal plasticity might be considered the final result of activation and/or inhibition of molecular cascades regulating the dynamics of the microtubular system. In addition, the prospect of targeting microtubules as a pharmacotherapeutic approach to treat mood disorders will be discussed. PMID- 16266293 TI - Bile-salt-stimulated lipase and mucins from milk of 'secretor' mothers inhibit the binding of Norwalk virus capsids to their carbohydrate ligands. AB - Breast-feeding-associated protection against calicivirus diarrhoea is associated with the presence of high levels of 2-linked oligosaccharides in mother's milk, and human calicivirus strains including the NV (Norwalk virus) use gut 2-linked fucosylated glycans as receptors, suggesting the presence of decoy receptors in milk. Our aim was to analyse the ability of human milk to inhibit the attachment of rNV VLPs (recombinant NV-like particles) to their carbohydrate ligands and to characterize potential inhibitors found in milk. Milk from women with the secretor phenotype was strongly inhibitory, unlike milk from women that are non secretors, which is devoid of 2-linked fucosylated structures. At least two fractions in human milk acted as inhibitors for the NV capsid attachment. The first fraction corresponded to BSSL (bile-salt-stimulated lipase) and the second to associated mucins MUC1 and MUC4. These proteins present tandem repeat O glycosylated sequences that should act as decoy receptors for the NV, depending on the combined mother/child secretor status. PMID- 16266296 TI - Of hearts, kidneys, . . . and artificial organ technologies. PMID- 16266295 TI - Gene expression patterns and catalytic properties of UDP-D-glucose 4-epimerases from barley (Hordeum vulgare L.). AB - UGE (UDP-Glc 4-epimerase or UDP-Gal 4-epimerase; EC 5.1.3.2) catalyses the interconversion of UDP-Gal and UDP-Glc. Both nucleotide sugars act as activated sugar donors for the biosynthesis of cell wall polysaccharides such as cellulose, xyloglucans, (1,3;1,4)-beta-D-glucan and pectins, together with other biologically significant compounds including glycoproteins and glycolipids. Three members of the HvUGE (barley UGE) gene family, designated HvUGE1, HvUGE2 and HvUGE3, have been characterized. Q-PCR (quantitative real-time PCR) showed that HvUGE1 mRNA was most abundant in leaf tips and mature roots, but its expression levels were relatively low in basal leaves and root tips. The HvUGE2 gene was transcribed at significant levels in all organs examined, while HvUGE3 mRNA levels were very low in all the organs. Heterologous expression of a near full length cDNA confirmed that HvUGE1 encodes a functional UGE. A non-covalently bound NAD+ was released from the enzyme after denaturing with aqueous ethanol and was identified by its spectrophotometric properties and by electrospray ionization MS. The K(m) values were 40 microM for UDP-Gal and 55 muM for UDP-Glc. HvUGE also catalyses the interconversion of UDP-GalNAc and UDP-GlcNAc, although it is not known if this has any biological significance. A three-dimensional model of the HvUGE revealed that its overall structural fold is highly conserved compared with the human UGE and provides a structural rationale for its ability to bind UDP-GlcNAc. PMID- 16266299 TI - In vivo conditioning of tissue-engineered heart muscle improves contractile performance. AB - The ability to engineer cardiac tissue in vitro is limited by the absence of a vasculature. In this study we describe an in vivo model which allows neovascularization of engineered cardiac tissue. Three-dimensional cardiac tissue, termed "cardioids," was engineered in vitro from the spontaneous delamination of a confluent monolayer of cardiac cells. Cardioids were sutured onto a support framework and then implanted in a subcutaneous pocket in syngeneic recipient rats. Three weeks after implantation, cardioids were recovered for in vitro force testing and histological evaluation. Staining for hematoxylin and eosin demonstrated the presence of viable cells within explanted cardioids. Immunostaining with von Willebrand factor showed the presence of vascularization. Electron micrographs revealed the presence of large amounts of aligned contractile proteins and a high degree of intercellular connectivity. The peak active force increased from an average value of 57 microN for control cardioids to 447 microN for explanted cardioids. There was also a significant increase in the specific force. There was a significant decrease in the time to peak tension and half relaxation time. Explanted cardioids could be electrically paced at frequencies of 1-5 Hz. Explanted cardioids exhibited a sigmoidal response to calcium and positive chronotropy in response to epinephrine. As the field of cardiac tissue engineering progresses, it becomes desirable to engineer larger diameter tissue equivalents and to induce angiogenesis within tissue constructs. This study describes a relatively simple in vivo model, which promotes the neovascularization of tissue-engineered heart muscle and subsequent improvement in contractile performance. PMID- 16266300 TI - End-point temperature of rewarming after hypothermic cardiopulmonary bypass in pediatric patients. AB - In an attempt to find an adequate end-point rewarming temperature after hypothermic cardiopulmonary bypass (CPB), 50 pediatric patients who underwent cardiac surgery were randomly assigned for the end-point rectal rewarming temperature at either 35.5 (Group 1) or 37.0 degrees C (Group 2). The patients' rectal temperature, with heart rate and blood pressure, was measured 0.5, 1.0, 4.0, 8.0, and 16.0 h after the arrival in the intensive care unit. For all patients, nonpulsatile perfusion with a roller pump and a membrane or bubble oxygenator was used for oxygenation. Age, sex, body surface area, total bypass time, and rewarming time were comparable in both groups. No afterdrop and no statistical differences in the rectal temperatures between the two groups were observed. Also, no statistical differences were observed between the two groups with respect to the heart rate and blood pressure. No shivering was noted in all patients. In conclusion, with the restoration of rectal temperature above 35.5 degrees C at the end of CPB in pediatric patients, the present study found no afterdrop. PMID- 16266294 TI - The family feud: turning off Sp1 by Sp1-like KLF proteins. AB - Sp1 is one of the best characterized transcriptional activators. The biological importance of Sp1 is underscored by the fact that several hundreds of genes are thought to be regulated by this protein. However, during the last 5 years, a more extended family of Sp1-like transcription factors has been identified and characterized by the presence of a conserved DNA-binding domain comprising three Kruppel-like zinc fingers. Each distinct family member differs in its ability to regulate transcription, and, as a consequence, to influence cellular processes. Specific activation and repression domains located within the N-terminal regions of these proteins are responsible for these differences by facilitating interactions with various co-activators and co-repressors. The present review primarily focuses on discussing the structural, biochemical and biological functions of the repressor members of this family of transcription factors. The existence of these transcriptional repressors provides a tightly regulated mechanism for silencing a large number of genes that are already known to be activated by Sp1. PMID- 16266301 TI - Thrombin-free fibrin coating on small caliber vascular prostheses has high antithrombogenicity in rabbit model. AB - Fibrin coatings on prosthetic vascular graft, which are conventionally produced by fibrinogen and thrombin, are expected to improve antithrombogenicity and healing characteristics. Thrombin is one of the factors of blood coagulation cascade; however, it has a possibility to play a negative role in the graft antithrombogenicity. The purpose of this study was to evaluate the performance of our new grafts, thrombin-free fibrin-coated small caliber vascular prostheses. Knitted polyester fabric vascular prostheses 2 mm in internal diameter were coated with fibrin coating with thrombin (Graft I) or without thrombin (Graft II). Both grafts were implanted in bilateral common carotid arteries of 35 Japanese white rabbits, with Graft I in one side and Graft II in the contralateral side. Graft patency, histology, thrombin activity, and platelet deposition were compared between both grafts on postoperative days (PODs) 1, 3, 7, 10, 14, 30, and 60. Both grafts were patent without thrombus or stenosis at each end point (maximal period, POD 60). Macro- and microscopic findings revealed that no obvious difference was observed between both grafts. Before graft implantation, thrombin activities in Grafts I and II were 0.711 +/- 0.086 and 0.009 +/- 0.007 optical density at 405 nm, respectively. Thrombin activity of Graft II was significantly less than that of Graft I in every period after graft implantation, and platelet deposition of Graft II was significantly less than that of Graft I until POD 30. Thrombin-free fibrin-coated vascular prostheses have superior performance of antithrombogenicity to conventional fibrin-coated vascular prostheses with thrombin. PMID- 16266302 TI - Effect of temperature increase on the distensibility of porcine thoracic aorta. AB - The objective of the study was to investigate the temperature impact on the elasticity of porcine thoracic aorta. Under general anesthesia, 16 Landrace pigs were subjected to thoracotomy, and the descending thoracic aorta was removed and stored in normal saline. Serial sections of the vessel created cylindrical aortic specimens which were tested in a uniaxial tension device to determine the elastic properties of the aortic wall. In the control, Group A (n = 8), the aortic tissues were tested while immersed in normal saline bath of temperature TA = 37.0 +/- 0.5 degrees C, while in Group B, the temperature was TB = 40.0 +/- 0.5 degrees C. Each experiment took place only after the tissues had remained for 15 min in temperature TA or TB. For the results, the stiffness modulus of Group B showed a significant decrease (P < 0.05) at medium strain level deformation (e = 1, SB1 = 114 +/- 8 Pa) as well as at high strain level deformation (e = 2, SB1 = 1182 +/- 48 Pa) in comparison with the control, Group A (e = 1, SA1 = 147 +/- 15 Pa; e = 2, SA1 = 1479 +/- 64 Pa). It is concluded that temperature increase facilitates, in vitro, the expansion of descending thoracic aorta. We assume that thermal treatment may be another means against the stiffening of aorta, which calls for further research. PMID- 16266303 TI - pO2 and pCO2 increment in post-dialyzer blood: the role of dialysate. AB - Blood returning from a dialyzer during hemodialysis has a higher pO2 and pCO2 content than blood entering the dialyzer, and this has been attributed to the dialysate. The present study investigates this phenomenon. Acid-base and blood gas parameters (pH, pO2, pCO2 and HCO3) were measured in three groups of stable chronic hemodialysis patients (A, B, and C) undergoing high-flux hemodialysis. In group A (n = 15), "arterial" (a) and "venous" (v) samples were withdrawn simultaneously before dialysis (samples A0), 5 min after circulation of the blood with the dialysate in the by-pass mode (samples A5), and 5 min after high-flux hemodialysis at a zero ultrafiltration rate (samples A10). In group B (n = 11) (a) and (v) samples were withdrawn simultaneously before dialysis (samples B0), 5 min after isolated-ultrafiltration with closed dialysate ports ("isolated-closed" ultrafiltration) (samples B5), and 5 min after high-flux hemodialysis at a zero ultrafiltration rate (samples B10). In group C (n = 14), after an initial arterial blood sample withdrawal before hemodialysis (sample C0), high-flux hemodialysis at a zero ultrafiltration rate was initiated. Five minutes later, blood and dialysate samples were withdrawn simultaneously from the hemodialysis lines (samples C5). In all cases blood and dialysate (bicarbonate) flow rates were set at 0.300 and 0.700 L/min, respectively. FLX-18 hemodialyzers (membrane PEPA 1.8 m2) were used in this study. Analysis of variance revealed significant changes only in venous samples. A comparison of arterial and venous samples revealed no differences between groups A and B before the initiation of dialysis (A0a vs. A0v and B0a vs. B0v, P = NS). The pO2 content was higher in A5v samples than in A5a samples (83.5 +/- 11.2 vs. 88.8 +/- 14.0 mm Hg, P < 0.02), while the level of HCO(3) was higher in A5a samples than in A5v samples (20.8 +/- 2.0 vs. 20.4 +/- 1.8 mEq/L, P < 0.05). A10a samples possessed a higher pH and lower levels of pO2, pCO2, and HCO3 in comparison to A10v samples (P < 0.001 for all). Mean pO2 and pCO2 values in A5v and A10v samples increased by 6.3% and 12.1% and by 1.29% and 52% in comparison to corresponding values of A5a and A10a samples, respectively. The pO2 level was the only parameter that differed significantly between B5a and B5v samples (B5a = 84.6 +/- 10.1 vs. B5v = 98.0 +/- 12.6 mm Hg, P < 0.005). B10a samples possessed a higher pH and lower levels of pCO2, pO2, and HCO3 in comparison to B10v samples (P < 0.0005 for all comparisons). Mean pO2 and pCO2 values in B5v and B10v samples increased by 16.2% and 16.3% and by -0.29% and 64.8% in comparison to corresponding values of B5a and B10a samples, respectively. C5a samples possessed a higher pH and lower levels of pCO2, pO2, and HCO3 in comparison to C5v samples (P < 0.001 for all). Mean pO2 and pCO2 values in C5v samples were, respectively, 16.0% and 65.0% higher than corresponding values of C5a samples. These results indicate that blood returning from the dialyzer after 5 min of high-flux hemodialysis has a higher pO2 and pCO2 than blood entering the dialyzer, and that this difference is due to O2 and CO2 transfer from the dialysate space into the blood. PMID- 16266304 TI - Attitude toward organ donation: a survey in Pakistan. AB - This study was conducted to get an insight into the knowledge, attitude, and practices of the people of Pakistan regarding organ donation (OD), and to understand the various factors that influence these. Systematic sampling was used to generate a sample size of 357. Data collection was carried out via a questionnaire in the outpatient units of our hospital. All kinds of medical and surgical patients were interviewed. Knowledge of ODs that occur in Pakistan and the rest of the world was found to be 65.5 and 72.8%, respectively. The percentage of those willing to donate their organ/s was 59.9%. Knowledge of OD was significantly associated with education and socioeconomic status. Willingness to donate had a significant association with gender. It also had a moderate association with education and socioeconomic status. Awareness of OD and the knowledge that OD can save lives was also significantly associated with the willingness to donate. PMID- 16266305 TI - Design of a perfusion bioreactor specific to the regeneration of vascular tissues under mechanical stresses. AB - The objective of this work was to design a bioreactor to stimulate the three dimensional regeneration of arterial tissue on a cylindrical scaffold with a methodological approach. Once seeded, the scaffold is perfused internally and the externally with culture medium with two independent perfusion systems at different flow rates. The horizontal position and the rotation of the construct ensure the uniformity of the arterial growth and of the endothelial cell spreading. During cell culture, the parameters, such as internal flow and stretching of the vessel, can evolve gradually from the fetal stage to the adult stage. The bioreactor will also be useful for investigating the influence of mechanical stresses and strains on the properties of mature arteries (rigidity, burst strength, adhesion of endothelial cells, etc.). PMID- 16266307 TI - Hepatocyte function in a radial-flow bioreactor using a perfluorocarbon oxygen carrier. AB - The aims of this study were, first, to indicate the metabolic activity of hepatocytes in a radial-flow polyurethane foam matrix bioreactor relative to monocultures, and second, to evaluate the effect on the hepatocytes of including a synthetic perfluorocarbon (PFC) oxygen carrier to the recirculating medium. The efficient O2-carrying ability of PFCs may be beneficial to bioreactors employed in stressed cellular environments. Thus, they may also be useful in the treatment of an acute liver failure patient with a bioartificial liver support system (BALSS). Data on the function of three-dimensional (3-D) hepatocyte cultures exposed to emulsified PFCs are lacking. RESULTS: the metabolic functions of the 3 D hepatocyte cultures were improved relative to monocultures. Three-dimensional cultures with and without PFC behaved similarly, and no adverse effects could be detected when PFC was included in the recirculating medium. The addition of PFC significantly improved lidocaine clearance possibly due to the presence of higher O2 tension in the medium. Imaging indicated that large aggregates formed and that seeding had followed flow through the matrix. Simulations indicated first, that the cell numbers used in this study had been insufficient to challenge the bioreactor O2 supply explaining the similarity in performance of the 3-D cultures, and second, that the benefit of adding PFC would be more pronounced at the cell densities likely to be used in a BALSS bioreactor. PMID- 16266306 TI - Development of the pulsation device for rotary blood pumps. AB - A rotary blood pump (RP) is desirable as a small ventricular assist device (VAD). However, an RP is nonpulsatile. We tried to develop a device that attaches a pulse to the RP. We also tried to develop a pulse-generating equipment that was not air-pressure driven. The ball screw motor was considered a candidate. The application of a small-sized shape memory alloy was also attempted. An electrohydraulic system was adopted, and actuator power was connected to the diaphragm. The diaphragm was placed on the outer side of the ventricle. Most RPs that have been developed all over the world drain blood from the ventricle. The wave of a pulse should be generated if a pulse is added by the drawn part. The output assistance from the outer side of the ventricle was attempted in animal experiments, and the device operated effectively. This device can be used during implantable operation of RP. This may serve as an effective device in patients experiencing problems in peripheral circulation and in the function of internal organs. PMID- 16266308 TI - Enhancement of the transmesothelial resistance of the parietal sheep peritoneum by epinephrine in vitro: ussing-type chamber experiments. AB - The peritoneal mesothelium constitutes an ion transport barrier that is taken advantage of in peritoneal dialysis. The aim of this study was to investigate the effects of epinephrine on the electrical transmesothelial resistance (R(TM)) of the isolated parietal sheep peritoneum by means of Ussing-type chamber experiments. Intact parietal (diaphragmatic) peritoneal samples were obtained from adult sheep immediately after sacrifice and transferred within 0.5 h to the laboratory in a cooled Krebs-Ringer bicarbonate solution (4 degrees C, pH 7.5), bubbled with 95% O2-5% CO2. A parietal peritoneal planar sheet was mounted in a Ussing-type chamber. Epinephrine (10(-7) M) was added to the apical and the basolateral side. The R(TM) was measured before and serially after the addition of epinephrine for 30 min. As active ion transport is temperature-dependent, all measurements were performed at 37 degrees C. The results were calculated as means with standard errors (x +/- SE) of six independent experiments. The control R(TM) was 20.05 +/- 0.61 ohm x cm2. The addition of epinephrine to the basolateral side within 1 min induced an increase of R(TM) to 21.8 +/- 0.45 ohm x cm2, which decreased thereafter progressively to reach control values again after 15 min. A similar effect of epinephrine on the apical side was apparent with a rapid rise of R(TM) to 22.5 +/- 0.66 ohm x cm2 and a subsequent decrease (P < 0.05). A clear association between the R(TM) and active ion transport was established from previous studies. The results of our study indicate a rapid action of epinephrine on the parietal peritoneum permeability. PMID- 16266309 TI - The future of human valve allografts: bioengineering and stem cells. PMID- 16266310 TI - Restoration of hepatic mast cells and expression of a different mast cell protease phenotype in regenerating rat liver after 70%-hepatectomy. AB - Mast cells (MC) can undergo significant changes in number and phenotype; these alterations result in the differential expression of growth factors and cytokines. Kit ligand (KL; stem cell factor) is produced by mesenchymal cells, and in the liver by biliary epithelial cells. Recent studies suggest that KL, and its receptor c-kit, may be involved in liver regeneration after loss of liver mass. However, KL is also the major growth, differentiating, chemotactic, and activating factor for MC. The aim of our study was to elucidate the dynamics and phenotype of hepatic MC and KL/c-kit expression during liver regeneration after partial (70%) hepatectomy in the rat. Regenerating livers were harvested after 1, 3, 7, and 14 days, respectively (n = 6 each day). MC were stained for naphthol AS/D-chloroacetate esterase and counted as MC per bile ductule. MC phenotype was assessed by rat MC protease (RMCP)-1 and -2 immunofluorescence staining, in order to distinguish RMCP-1 positive connective tissue MC (CTMC) from RMCP-2 positive mucosa MC (MMC). mRNA expression of RMCP, c-kit, and the differentially spliced variants of KL was quantified by RT-PCR. MC counts per bile ductule decreased in regenerating rat liver tissue at day 3, compared with native livers, and became normal thereafter. Hepatic MC were predominantly of a CTMC phenotype expressing RMCP-1, as previously published; after hepatectomy, between 76 and 99% of all MC double-expressed RMCP-1 and -2, compatible with an MMC phenotype. The ratio of the two alternatively spliced mRNAs for KL (KL-1 : KL-2), and c-kit mRNA expression did not differ significantly between regenerating livers and the livers of sham operated animals. These results suggest that hepatic mast cells are restored during liver regeneration after partial hepatectomy in the rat. Restored MC express an MMC phenotype, suggesting migration from outside into the regenerating liver. Alternative splicing of KL is affected by the surgical procedure in general, and, together with its receptor c-kit, doesn't seem to be involved in liver regeneration after partial hepatectomy in the rat. Further functional studies, and studies in regenerating human livers might offer the possibility of elucidating the role of the hepatic mast cell, and its different protease phenotypes during liver regeneration after surgical loss of liver mass. PMID- 16266311 TI - Lymphocytes and MHC class II positive cells in the female rabbit reproductive tract before and after ovulation. AB - In this study, we identified lymphocytes and MHC class II positive (MHC-II+) cells in the reproductive tract of female rabbits both before and after ovulation. CD43+ T cells were frequently present in the mucosa of the oviduct, cervix, and vagina, but far fewer positive cells were seen in the endometrium. The induction of ovulation did not change the cell density in these regions. KEN 5+ T cells and MHC-II+ cells were also frequently seen in the mucosa of the oviduct, cervix, and vagina both before and after ovulation. However, in the uterus, there were very few positive cells before ovulation, but the number increased dramatically after ovulation. Associated with the increase of KEN-5+ T cells, IL-2 mRNA expression in the uterus also increased after ovulation, suggesting that the uterus experienced an increase of T-cell activation. IgM- and IgA-positive B cells were not commonly seen in the reproductive tract and the induction of ovulation did not alter this. Our results suggest that the reproductive tract of female rabbits has the capacity to mount an immune response and that the immune cell distribution of the rabbit reproductive tract has some distinctive features compared with that found in other species. PMID- 16266312 TI - Role of oxidative stress in ERK and p38 MAPK activation induced by the chemical sensitizer DNFB in a fetal skin dendritic cell line. AB - The intracellular mechanisms involved in the early phase of dendritic cell (DC) activation upon contact with chemical sensitizers are not well known. The strong skin sensitizer 2,4-dinitrofluorobenzene (DNFB) was shown to induce the activation of mitogen-activated protein kinases (MAPK) in DC. In the present study, we investigated a putative role for oxidative stress in DNFB-induced MAPK activation and upregulation of the costimulatory molecule CD40. In a DC line generated from fetal mouse skin, DNFB induced a significant increase in protein oxidation, measured by the formation of carbonyl groups, while it had almost no effect on lipid peroxidation. The antioxidants glutathione and vitamin E, which inhibit protein and lipid oxidation, respectively, were used to assess the role of oxidative stress in DNFB-induced MAPK activation. Glutathione, but not vitamin E, inhibited DNFB-induced p38 MAPK and ERK1/2 phosphorylation, whereas none of the antioxidants interfered significantly with the DNFB-induced upregulation of CD40 protein levels. Taken together, these results indicate that DNFB activates p38 MAPK and ERK1/2 via production of reactive oxygen species, and that protein oxidation plays an important role in MAPK activation. PMID- 16266314 TI - Macrophage inflammatory protein-1alpha expression plasmid enhances DNA vaccine induced immune responses against HSV-2. AB - In this study, we examined the effectiveness of macrophage inflammatory protein (MIP)-1alpha cDNA as a HSV-2 DNA vaccine adjuvant. pcDNA3-gD (pgD) and pcDNA3-MIP 1alpha (pMIP-1alpha) were co-injected to examine the modulatory effects of MIP 1alpha on immune phenotype and protection against lethal challenge with HSV-2. We found that Th-cell proliferative responses were dramatically enhanced by co injection of pgD and pMIP-1alpha compared with injection of pgD alone. The secretion of IL-2 and IFN-gamma was also significantly increased by pgD and pMIP 1alpha co-injection; however, the production of cytokines IL-4 and IL-10 was not affected by co-injection. pgD and pMIP-1alpha co-injection resulted in a moderate enhancement of systemic gD-specific antibody level, but mucosal secretory IgA was markedly enhanced. When BALB/c mice were challenged intravaginally with 100 LD50 of HSV-2 strain Sav, pMIP-1alpha co-injection with pgD improved their survival rate and significantly reduced both the number of mice with lesions and the lesion severity. Therefore, MIP-1alpha cDNA as a HSV-2 DNA vaccine adjuvant drives antigen-specific Th1-type responses, reducing HSV-2-derived morbidity and mortality. PMID- 16266313 TI - Recombinant fowlpox virus for in vitro gene delivery to pancreatic islet tissue. AB - The feasibility of using avipox virus as a vector for gene delivery to islet tissue (adult islets and fetal proislets) was examined using a recombinant fowlpox virus (FPV) engineered to express the reporter gene LacZ (FPV-LacZ). The efficiency of in vitro transduction was dose-dependent and influenced by the donor species and maturation status of the islet tissue. Reporter gene expression in FPV-LacZ-transduced islet grafts was transient (3-7 days) in immunoincompetent nude mice and was not prolonged by in vivo treatment with anti-IFN-gamma mAb. In contrast, FPV-LacZ-transduced NIT-1 cells (a mouse islet beta cell line) expressed the LacZ gene beyond 18 days in vitro. Silencing of transgene expression therefore appeared to occur in vivo and was T cell- and IFN-gamma independent. Isografts of FPV-LacZ-transduced islets in immunocompetent mice underwent immunological destruction by 7 days, suggesting that either FPV proteins or the reporter protein beta-galactosidase induced an adaptive immune response. Co-delivery of the rat bioactive immunoregulatory cytokine gene TGF beta to islets using FPV-TGF-beta led to enhanced expression of TGF-beta mRNA in isografts but no long-term protection. Nevertheless, compared to control islet isografts at 5 days, FPV-transduced islets remained embedded in the clotted blood used to facilitate implantation. This phenomenon was TGF-beta transgene independent, correlated with lack of cellular infiltration, and suggested that the FPV vector transformed the blood clot into a temporary immunological barrier. PMID- 16266315 TI - Detection of prion epitopes on PrP and PrP of transmissible spongiform encephalopathies using specific monoclonal antibodies to PrP. AB - Amino acid residues 90-120 of the prion protein (PrP) are likely to be critical for the conversion of PrP(c) to PrP(sc) in the transmissible spongiform encephalopathies. We raised 10 monoclonal antibodies against the 90-120 amino acid region, mapped the epitope specificity of these anti-PrP antibodies, and investigated the expression of epitopes recognized by the antibodies in both PrP(c) and PrP(sc). Four out of five of the anti-PrP antibodies raised in a prion knockout mouse immunized with the linear peptide of PrP90-120 could detect PrP(sc) in 'native' and denatured forms and PrP(c) in normal cells, as well as recognize epitopes within PrP93-112 residues. In contrast, the other six anti-PrP reagents, including five raised from the two knockout mice immunized with conformationally modified PrP90-120 peptide, could detect PrP(c) and recognize epitopes within PrP93-107 residues. Four of these reagents could also detect denatured PrP(sc) on western blots but not PrP(sc) plaques in brain tissue. The results indicate that residues PrP93-102 are exposed in PrP(c) but are buried upon conversion to the PrP(sc) isoform. Furthermore, PrP103-107 residues are partially buried in PrP(sc) while only the PrP107-112 epitope remains exposed, suggesting that the region PrP93-112 undergoes conformational changes during its conversion to PrP(sc). PMID- 16266316 TI - Protection against malaria by anti-erythropoietin antibody due to suppression of erythropoiesis in the liver and at other sites. AB - We have previously reported that erythropoiesis commences in the liver and spleen after malarial infection, and that newly generated erythrocytes in the liver are essential for infection of malarial parasites as well as continuation of infection. At this time, erythropoietin (EPO) is elevated in the serum. In the present study, we administered EPO or anti-EPO antibody into C57BL/6 (B6) mice to modulate the serum level of EPO. When mice were infected with a non-lethal strain (17NXL) of Plasmodium yoelii (blood-stage infection of 10(4) parasitized erythrocytes per mouse), parasitemia continued for 1 month, showing a peak at day 17. Daily injection of EPO (200 IU/day per mouse) from day five to day 14 prolonged parasitemia, whereas injection of anti-EPO antibody (1.5 mg/day per mouse) every second day from day five to day 28 decreased it. Erythropoiesis was confirmed in the liver, spleen and bone marrow by the appearance of nucleated erythrocytes (TER119+). When anti-EPO antibody was injected by the same protocol into mice infected with a lethal strain (17XL) of P. yoelii, all mice showed decreased parasitemia and recovered from the infection. These results suggest that the use of anti-EPO antibody after malarial infection may be of therapeutic value in severe cases of malaria. PMID- 16266317 TI - IFN-gamma enhances bovine macrophage responsiveness to Mycobacterium bovis: Impact on bacterial replication, cytokine release and macrophage apoptosis. AB - We sought to determine the impact of bovine IFN-gamma on the interaction between Mycobacterium bovis and bovine macrophages. Bovine macrophages released small amounts of nitric oxide (NO), TNF-alpha, IL-1beta and IL-12 upon infection with bacille Calmette-Guerin (BCG). Prior pulsing of cells with IFN-gamma significantly enhanced the release of NO and IL-12. Infection of bovine macrophages with virulent M. bovis led to the release of higher levels of pro inflammatory mediators, compared to levels released upon BCG infection. IFN-gamma treatment of macrophages enhanced the release of pro-inflammatory mediators, but did not modify bacterial replication in M. bovis-infected macrophages. Treatment of macrophages with a combination of IFN-gamma and LPS led to a reduction in bacterial replication. Infected cells treated with IFN-gamma/LPS progressed mostly through an apoptotic pathway, whereas untreated infected cells eventually died by necrosis. Agents that prevented the acquisition of bacteriostatic activity by activated macrophages also prevented the induction of apoptosis in infected macrophages (IL-10 and neutralizing anti-TNF-alpha). We conclude that virulent M. bovis is a major determinant of release of pro-inflammatory cytokines by macrophages. IFN-gamma amplifies the macrophage cytokine release in response to M. bovis. Induction of apoptosis is closely linked to the emergence of macrophage resistance to M. bovis replication, which is dependent on endogenous TNF-alpha release. PMID- 16266318 TI - Evidence that insulin-like growth factor-1 requires protein kinase C-epsilon, PI3 kinase and mitogen-activated protein kinase pathways to protect human vascular smooth muscle cells from apoptosis. AB - Insulin-like growth factor (IGF)-1 has been implicated in the development of occlusive vascular lesions. Although its role in vascular smooth muscle cell (VSMC) growth and migration are fairly well characterized, anti-apoptotic signals of IGF-1 in human VSMC remain largely unknown. In this study, we examined IGF-1 signals that protect human and rat VSMC from staurosporine (STAU)- and c-myc- induced apoptosis, respectively. Treatment with STAU resulted in apoptotic DNA fragmentation, phosphatidylserine externalization and cell shrinkage, but only occasional VSMC 'blebbing'. STAU-induced death and IGF-1-mediated survival were concentration dependent, while time-lapse video microscopy showed that IGF-1 inhibited c-myc-induced apoptosis by 90%. Pretreatment with mitogen-activated protein kinase/extracellular signal regulated kinase kinase (MEK) inhibitors UO126 and PD098059, or with the phosphatidylinositol 3-kinase (PI3-K) inhibitor wortmannin, reversed IGF-1-mediated human VSMC survival by 25-27% and 66%, respectively. Translocation studies showed that IGF-1 activated protein kinase C (PKC)-epsilon, but not PKC-alpha or PKC-delta, even in the presence of STAU, while pharmacological PKC inhibition (Ro-318220 or Go6976) implicated PKC-zeta or a novel PKC isozyme in IGF-1-mediated survival. Transient expression of activated PKC-epsilon but not activated PKC-zeta decreased myc-induced apoptosis in rat VSMC. In human VSMC, antisense oligodeoxynucleotides to PKC-epsilon partially reversed IGF-1-induced survival. In addition, IGF-1 elicited a mild but sustained activation of extracellular signal regulated kinase (ERK)1/2 in human VSMC that was abolished after 1 h in the presence of STAU. PKC downregulation reversed both IGF-1- and PMA-induced ERK activity, but platelet-derived growth factor (PDGF) induced activity was unchanged. These results indicate for the first time that IGF-1 can protect human VSMC via multiple signals, including PKC-epsilon, PI3-K and mitogen-activated protein kinase pathways. PMID- 16266319 TI - Hypoxia inhibits the migratory capacity of human monocyte-derived dendritic cells. AB - Hypoxia, a prominent characteristic of inflammatory tissue lesions and solid tumour microenvironments, is a crucial stimulus capable of modulating the expression of specific genes involved in leucocyte recruitment. Although studies have shown that hypoxia can affect leucocyte migration by influencing the expression of migration-related genes, such as matrix metalloproteinases (MMP) and their endogenous tissue inhibitors of matrix metalloproteinases (TIMP), it remains unclear whether hypoxia can affect the migration of dendritic cells (DC). In this study, we showed that human monocyte-derived DC under hypoxic conditions in a transwell system have significantly reduced migratory capacity compared to normoxic controls. A moderate phenotypic change of hypoxic DC was observed. In hypoxic DC, we detected a twofold increase in TIMP-1 transcript levels, and downregulated expression of MMP-9 and membrane type 1-MMP genes by threefold and 17-fold, respectively. Our results suggest that hypoxia may inhibit DC migratory activity by regulating the balance between MMP and TIMP gene expression. PMID- 16266320 TI - The alpha-gal epitope and the anti-Gal antibody in xenotransplantation and in cancer immunotherapy. AB - The alpha-gal epitope (Galalpha1-3Galbeta1-(3)4GlcNAc-R) is abundantly synthesized on glycolipids and glycoproteins of non-primate mammals and New World monkeys by the glycosylation enzyme alpha1,3galactosyltransferase (alpha1,3GT). In humans, apes and Old World monkeys, this epitope is absent because the alpha1,3GT gene was inactivated in ancestral Old World primates. Instead, humans, apes and Old World monkeys produce the anti-Gal antibody, which specifically interacts with alpha-gal epitopes and which constitutes approximately 1% of circulating immunoglobulins. Anti-Gal has functioned as an immunological barrier, preventing the transplantation of pig organs into humans, because anti-Gal binds to the alpha-gal epitopes expressed on pig cells. The recent generation of alpha1,3GT knockout pigs that lack alpha-gal epitopes has resulted in the elimination of this immunological barrier. Anti-Gal can be exploited for clinical use in cancer immunotherapy by targeting autologous tumour vaccines to APC, thereby increasing their immunogenicity. Autologous intact tumour cells from haematological malignancies, or autologous tumour cell membranes from solid tumours are processed to express alpha-gal epitopes by incubation with neuraminidase, recombinant alpha1,3GT and with uridine diphosphate galactose. Subsequent immunization with such autologous tumour vaccines results in in vivo opsonization by anti-Gal IgG binding to these alpha-gal epitopes. The interaction of the Fc portion of the vaccine-bound anti-Gal with Fcgamma receptors of APC induces effective uptake of the vaccinating tumour cell membranes by the APC, followed by effective transport of the vaccinating tumour membranes to the regional lymph nodes, and processing and presentation of the tumour-associated antigen (TAA) peptides. Activation of tumour-specific T cells within the lymph nodes by autologous TAA peptides may elicit an immune response that in some patients will be potent enough to eradicate the residual tumour cells that remain after completion of standard therapy. A similar expression of alpha-gal epitopes can be achieved by transduction of tumour cells with an adenovirus vector (or other vectors) containing the alpha1,3GT gene, thus enabling anti-Gal-mediated targeting of the vaccinating transduced cells to APC. Intratumoral delivery of the alpha1,3GT gene by various vectors results in the expression of alpha-gal epitopes. Such expression of the xenograft carbohydrate phenotype is likely to induce anti-Gal-mediated destruction of the tumour lesion, similar to rejection of xenografts by this antibody. Opsonization of the destroyed tumour cell membranes by anti-Gal IgG further targets them to APC, thus converting the tumour lesion, treated by the alpha1,3GT gene, into an in situ autologous tumour vaccine. PMID- 16266321 TI - Alpha1,3-galactosyltransferase knockout pigs are available for xenotransplantation: are glycosyltransferases still relevant? AB - In the early 1990s, the Galalpha(1,3)Gal carbohydrate linkage was found to be the major xenoepitope causing hyperacute rejection. This carbohydrate, the antibodies that bind to it, and the enzyme that produces it (alpha1,3-galactosyltransferase) were the foci of research by many groups. Nearly a decade later, alpha1,3 galactosyltransferase knockout pigs were finally produced; hyperacute rejection could be avoided in these pigs. Having achieved this goal, enthusiasm declined for the study of glycosyltransferases and their carbohydrate products. To examine whether this decline was premature, we evaluate whether gene deletion has indeed solved the initial rejection problem or, in fact, created new problems. This review addresses this by examining the impact of the gene deletion on cell surface carbohydrate. Surprisingly, Galalpha(1,3)Gal is still present in alpha1,3 galactosyltransferase knockout animals: it is possibly synthesized on lipid by iGb3 synthase. Furthermore, removal of alphaGal resulted in the exposure of the N acetyllactosamine epitope. This exposed epitope can bind natural antibodies and perhaps should be capped by transgenic expression of another transferase. We believe the continued study of glycosyltransferases is essential to examine the new issues raised by the deletion of alpha1,3-galactosyltransferase. PMID- 16266322 TI - Characteristics of protein-carbohydrate interactions as a basis for developing novel carbohydrate-based antirejection therapies. AB - The relative shortage of human organs for transplantation is today the major barrier to a broader use of transplantation as a means of treating patients with end-stage organ failure. This barrier could be partly overcome by an increased use of blood group ABO-incompatible live donors, and such trials are currently underway at several transplant centres. If xenotransplantation can be used clinically in the future, the human organ shortage will, in principle, be eradicated. In both these cases, carbohydrate antigens and the corresponding anti carbohydrate antibodies are the major primary immunological barriers to overcome. Refined carbohydrate-based therapeutics may permit an increased number of ABO incompatible transplantations to be carried out, and may remove the initial barriers to clinical xenotransplantation. Here, we will discuss the chemical characteristics of protein-carbohydrate interactions and outline carbohydrate based antirejection therapies as used today in experimental as well as in clinical settings. Novel mucin-based adsorbers of natural anti-carbohydrate antibodies will also be described. PMID- 16266323 TI - Three-dimensional structures of carbohydrate determinants of Lewis system antigens: implications for effective antibody targeting of cancer. AB - Lewis system carbohydrate antigens have been shown to be expressed at high levels in many cancers of epithelial cell origin, including those of colon, breast, lung, prostate and ovary. The type 1 (Le(a) and Le(b)) antigens are important histo-blood groups, while type 2 (Le(x) and Le(y)) antigens in healthy individuals are only expressed, at relatively low levels, by a few tissues, including some epithelial cells. Thus, the type 2 antigens are considered to be tumour-associated antigens and are promising targets for cancer treatment, including antibody-based immunotherapy. In this review, we discuss the conformational characteristics of the free and bound forms of Lewis oligosaccharides and the 3D structures of antibodies in complex with Le(y) and Le(x) antigens. Collectively, the structural studies have demonstrated that the Lewis determinants are rigid structures, which generally maintain the same conformation in the free and bound states. The rigid nature and similarities in shape of type 1 and 2 Lewis oligosaccharides appear to make them perfectly suited to driving a structurally convergent immune response (at least in the case of Le(y) specific antibodies) toward a highly specific recognition of individual carbohydrate determinants, which is a goal in the development of effective antibody-based cancer treatments. PMID- 16266325 TI - Viral haemorrhagic septicaemia virus in marine fish and its implications for fish farming--a review. AB - Viral haemorrhagic septicaemia virus (VHSV) has, in recent decades, been isolated from an increasing number of free-living marine fish species. So far, it has been isolated from at least 48 fish species from the northern hemisphere, including North America, Asia and Europe, and fifteen different species including herring, sprat, cod, Norway pout and flatfish from northern European waters. The high number of VHSV isolations from the Baltic Sea, Kattegat, Skagerrak, the North Sea and waters around Scotland indicate that the virus is endemic in these waters. The VHSV isolates originating from wild marine fish show no to low pathogenicity to rainbow trout and Atlantic salmon, although several are pathogenic for turbot. Marine VHSV isolates are so far serologically indistinguishable from freshwater isolates. Genotyping based on VHSV G- and N-genes reveals four groups indicating the geographical origin of the isolates, with one group representing traditional European freshwater isolates and isolates of north European marine origin, a second group of marine isolates from the Baltic Sea, a third group of isolates from the North Sea, and a group representing North American isolates. Examples of possible transfer of virus from free-living marine fish to farmed fish are discussed, as are measures to prevent introduction of VHSV from the marine environment to aquaculture. PMID- 16266326 TI - Interactions of microorganisms isolated from gilthead sea bream, Sparus aurata L., on Vibrio harveyi, a pathogen of farmed Senegalese sole, Solea senegalensis (Kaup). AB - Four bacterial isolates from farmed gilthead sea bream, Sparus aurata, included in a previous study as members of the Vibrionaceae and Pseudomonodaceae and the genus Micrococcus, have been evaluated for their adhesive ability to skin and intestinal mucus of farmed Senegalese sole, Solea senegalensis, and their antagonistic effect on Vibrio harveyi, a pathogen of sole. These isolates showed higher adhesion to sole mucus than the pathogenic strains of V. harveyi assayed. Only two of the isolates showed antagonistic activity to V. harveyi. Interactions of the four isolates with V. harveyi in respect of adhesion to skin and intestinal mucus under exclusion, competition and displacement conditions were studied. Three isolates were able to reduce the attachment to skin and intestinal sole mucus of a pathogenic strain of V. harveyi under displacement and exclusion conditions, but not under competition conditions. The in vivo probiotic potential of isolate Pdp11 was assessed by oral administration followed by challenge with the pathogenic V. harveyi strain Lg14/00. A group of 50 Senegalese sole received a commercial diet supplemented with 10(8) cfu g(-1) of lyophilized Lg14/00 for 15 days. A second group of fish received a non-supplemented commercial diet. After challenge the mortality of the fish receiving the diet supplemented with the potential probiotic isolate was significantly lower than that in the fish receiving the non-supplemented commercial diet. This study has shown that the ability to interfere with attachment of pathogens, as well as the adhesion to host surfaces, are suitable criteria for selection of candidate probiotics for use in the culture of Senegalese sole. PMID- 16266327 TI - Environmental and physiological conditions affecting Tetrahymena sp. infection in guppies, Poecilia reticulata Peters. AB - Parasitic infections caused by Tetrahymena sp. constitute a serious problem in guppies, Poecilia reticulata. Tetrahymena was isolated from skin lesions of naturally infected guppies in a commercial aquaculture farm, cultured in vitro and used in subsequent experimental infections. In addition to guppies, angelfish, Pterophyllum scalare, platyfish, Xiphophorus maculates, and neontetra, Paracheirodon innesi, were susceptible, whereas tilapia (Oreochromis niloticus xO. aureus) was resistant. The ciliate had a high affinity for dead fish. Skin abrasion did not affect the infection, but fish with gas bubble disease exhibited a significantly higher infection than non-affected fish. Infection was significantly higher when fish were exposed to high levels of ammonia, high organic load and low water temperatures. Under shipment conditions, infection was significantly elevated. Full recovery was achieved at a low fish density. Results suggest that poor environmental and physiological conditions enhance infection with Tetrahymena sp. PMID- 16266330 TI - Comparative WSSV infection routes in the shrimp genera Marsupenaeus and Palaemon. PMID- 16266328 TI - Development of a rapid assay for the diagnosis of Myxobolus cerebralis in fish and oligochaetes using loop-mediated isothermal amplification. AB - A loop-mediated isothermal amplification assay was developed for the rapid detection of Myxobolus cerebralis in both fish and oligochaete hosts. The assay was optimized to amplify parasitic DNA by incubation with Bst DNA polymerase and a set of six specially constructed primers at 65 degrees C for 60 min. The amplification products were detected visually using SYBR Green I dye which gave identical results to gel electrophoresis analysis. Parasite DNA was detected from infected oligochaetes, and from the anal fin, caudal fin, dorsal fin and operculum of clinically infected fish. This 'Myxo-LAMP' assay has a detection limit similar to that of a polymerase chain reaction assay (10(-6)), but is more rapid and only requires a water bath for amplification and is therefore practical for simple and rapid diagnosis of infected tissue. PMID- 16266329 TI - Detection and identification of Flavobacterium psychrophilum from gill washings and benthic diatoms by PCR-based sequencing analysis. AB - A nested polymerase chain reaction (PCR) amplification technique was used to detect Flavobacterium psychrophilum from washings of fish gill surfaces and benthic diatoms as environmental samples. Gill washing samples were prepared from kawamutsu, Zacco temminckii (Temminck & Schlegel) and oikawa, Z. platypus (Temminck & Schlegel). Benthic diatom samples were collected from stone surfaces. All samples were collected from rivers in Wakayama Prefecture, Japan from November 2003 to January 2004. Following simple DNA extraction using a chelating resin, nested PCR techniques targeting 16S-rDNA and gyrB regions were performed, and PCR products were cloned and sequenced. With nested PCR amplification for the 16S-rDNA gene, ambiguous PCR products were detected from two of six samples, and by cloning and sequencing analysis were found not to be DNA fragments amplified from F. psychrophilum. Using nested PCR for the gyrB gene, however, five of six samples were clearly positive for F. psychrophilum in agarose gel electrophoresis, and were found to be identical with nucleotide sequences of F. psychrophilumgyrB deposited in DNA databases by sequencing analysis. Results indicate that nested PCR for the gyrB region is a useful technique to detect low levels of F. psychrophilum from environmental samples contaminated with many other organisms. PMID- 16266332 TI - Stem cells: classifications, controversies, and clinical applications. AB - The application of stem cells in regenerative and reparative therapies is emerging in surgery. Published information can lead to an over simplified view of stem cells with respect to their definitions, tissues of origin, abilities to differentiate into tissue lineages, and their capacity for functional tissue regeneration. The goals of this review article are to define embryonic and adult stem cells, compare differences between them, and summarize their potential clinical applications. PMID- 16266333 TI - Prognostic factors in treating antebrachial growth deformities with a lengthening procedure using a circular external skeletal fixation system in dogs. AB - OBJECTIVE: To evaluate treatment of antebrachial growth deformities (AGD) with a lengthening procedure using a circular external skeletal fixation (CESF) system and to determine prognostic factors. STUDY DESIGN: Prospective clinical study. ANIMALS: Thirty-four dogs with unilateral AGD. METHODS: Length deficits, angular and rotational deformities, elbow incongruity (EI), osteoarthritis (OA) of the elbow and carpal joint, function, and cosmesis were determined before and after a CESF lengthening procedure. RESULTS: On admission, EI (21 dogs; 62%), OA of the elbow joint (17 dogs; 50%), carpal OA (12 dogs; 35%), and concomitant elbow and carpal OA (5 dogs; 7%) were common findings. Treatment significantly improved function (normal, 20 dogs; 60%) and cosmesis (normal, 22 dogs; 65%). Angular and rotational deformities were almost completely corrected with small remaining length deficits. Elbow and carpal OA increased significantly during the follow-up period. Significant correlations were demonstrated between initial elbow OA and final function (R=0.42, P=.02), initial function and final function (R=0.41, P=.02), and initial ulnar and radial deficit and final cosmesis (R=0.58, P=.0001 and R=0.45, P=.008). CONCLUSIONS: Treatment of AGD by a CESF lengthening procedure was successful despite small remaining length deficits. Initial elbow OA, function, and ulnar and radial length deficits are prognostic factors in the treatment of AGD. CLINICAL RELEVANCE: Initial elbow OA and initial function are prognostic factors in predicting the functional outcome of treatment of AGD with a CESF lengthening procedure in the dog. PMID- 16266334 TI - A modified lateral approach to the canine cervical spine: procedural description and clinical application in 16 dogs with lateralized compressive myelopathy or radiculopathy. AB - OBJECTIVE: To describe a modified lateral surgical approach to the cervical spine in dogs and evaluate clinical outcomes of dogs with neurologic disorders treated with this technique. STUDY DESIGN: Retrospective study. SAMPLE POPULATION: Sixteen dogs with lesions involving the cervical spine. METHODS: Medical records (1998-2002) of dogs that had a modified lateral approach to the cervical spine were reviewed. To reduce procedural morbidity, the surgical approach was modified from original descriptions of the technique by minimizing disruption of epaxial and extrinsic thoracic limb musculature and limiting the size of the operative field to the affected vertebral segment. Signalment, neurologic status on admission; onset, progression, and duration of clinical signs; diagnostic testing, etiologic diagnosis, surgical site(s), intra- and postoperative complications, hospitalization, time to optimum recovery, neurologic status at discharge, final neurologic status, and outcome were recorded. Neurologic status (0-5) was scored preoperatively, 1 day postoperatively, at discharge, and at recheck examination (8 dogs). Telephone interviews were used to obtain additional follow-up information. RESULTS: None of the dogs had postoperative deterioration in neurologic status. Outcomes were good or excellent in dogs with intervertebral disc disease, 3/4 dogs with caudal cervical spondylomyelopathy, 1/2 dogs with spinal neoplasia, and in 1 dog with the vascular anomaly; long-term outcomes were unknown in 2 dogs. Intraoperative complications occurred in 3 dogs and included controllable venous plexus hemorrhage (2) and incorrect lesion localization (1). One dog was euthanatized because of postoperative complications. Hospitalization, time to optimal recovery, and overall outcome were not different from previously reported results using other surgical approaches to treat analogous neurologic conditions. CONCLUSIONS: A modified lateral approach to the cervical spine is viable for surgical treatment of cervical myelopathic or radiculopathic lesions when exposure to foraminal and lateralized lesions of the vertebral canal involving the C2-C7 vertebral articulations is desirable. CLINICAL RELEVANCE: A modified lateral approach to the cervical spine can be successfully used in dogs of all sizes to treat caudal cervical spondylomyelopathy, other anomalous conditions of the cervical spine, intervertebral disc disease, and spinal neoplasms. Although long-term follow-up was not available for all patients, outcomes were generally favorable. PMID- 16266335 TI - Accuracy and optimization of force platform gait analysis in Labradors with cranial cruciate disease evaluated at a walking gait. AB - OBJECTIVE: To determine the combination of ground reaction forces (GRFs) that best discriminates between lame and non-lame dogs. To compare the sensitivity of force platform gait analysis and visual observation at detecting gait abnormalities in Labradors after surgery for rupture of the cranial cruciate ligament (CCL). ANIMALS: All dogs were adult Labrador Retrievers: 17 free of orthopedic and neurologic abnormalities, 100 with unilateral CCL rupture, and 131 studied 6 months after surgery for unilateral CCL injury, 15 with observable lameness. PROCEDURE: Dogs were walked over a force platform with GRF recorded during the stance phase. Analytic properties of force platform gait analysis were calculated for several combinations of forces. The probability of visual observation detecting a gait abnormality was compared with that of force platform gait analysis. RESULTS: We determined that a combination of peak vertical force (PVF) and falling slope were optimal for discriminating sound and lame Labradors. After surgery, many dogs (75%) with no observable lameness failed to achieve GRFs consistent with sound Labradors. CONCLUSION: A force platform is an accurate method of assessing lameness in Labradors with CCL rupture and is more sensitive than visual observation. Assessing lameness with a combination of GRFs is better than using univariate GRFs. CLINICAL RELEVANCE: Therapies for stifle lameness can be accurately and objectively evaluated using 2 vertical ground reaction forces obtained from a force platform. PMID- 16266336 TI - Force plate analysis before and after dorsal decompression for treatment of degenerative lumbosacral stenosis in dogs. AB - OBJECTIVE: Using force plate analysis (FPA), determine ground reaction forces in dogs with degenerative lumbosacral stenosis (DLS) and evaluate the effects of lumbosacral decompressive surgery. STUDY DESIGN: Prospective clinical study. ANIMALS: Twelve dogs with DLS. METHODS: DLS was diagnosed by clinical signs, radiography, computed tomography, and/or magnetic resonance imaging. FPA was performed before surgery, and 3 days, 6 weeks, and 6 months after surgery. The mean peak braking (Fy+), peak propulsive (Fy-), and peak vertical (Fz+) forces of 8 consecutive strides were determined. The ratio between the total Fy- of the pelvic limbs and the total Fy- of the thoracic limbs (P/TFy-), reflecting the distribution of Fy-, was analyzed to evaluate any changes in locomotion pattern postoperatively. Ground reaction force data for DLS dogs were compared with data derived from 24 healthy dogs (control). RESULTS: In dogs with DLS, the propulsive forces (Fy-) of the pelvic limbs were significantly smaller than those of controls. P/TFy- was significantly smaller in dogs with DLS than in control dogs, and increased during the follow-up period, reaching normal values 6 months after surgery. CONCLUSIONS: Cauda equina compression in dogs with DLS decreases the propulsive force of the pelvic limbs and surgical treatment restores the propulsive force of the pelvic limbs in a 6-month period. CLINICAL RELEVANCE: In dogs with DLS, FPA is an effective method in evaluating the response to surgical treatment. Normal propulsive force in the pelvic limbs was restored during 6 months after decompressive surgery. PMID- 16266337 TI - Comparison of computed tomographic and standard radiographic determination of tibial torsion in the dog. AB - OBJECTIVE: To compare the effect of internal tibial rotation on the computed tomographic (CT) and standard radiographic assessment of tibial torsion (TT) in dogs. STUDY DESIGN: In vitro study. SAMPLE POPULATION: Cadaveric canine hind limbs (6 pairs). METHODS: The cranial cruciate ligament was transected, and caudo cranial radiographic and transverse CT images were obtained with the femur and tibiae in a neutral position, and after 15 degrees internal tibial rotation. Radiographic TT was determined by measuring the distance (d) between the calcaneus and the sulcus of the talus. CT determination of TT was performed using the proximal transcondylar and the distal cranial tibial axes. The distance (d) in the 2 groups and the difference in the CT determination of TT between groups were compared with a hypothetical mean value of 0 mm and 0 degrees, respectively. RESULTS: The mean distance (d) for the neutral radiographic group was not significantly different from 0 (P=.473); however, for the 15 degree group it was significantly different (P<.0001). The difference in the CT determination of TT did not differ from 0 (P=.317). CONCLUSION: The standard radiographic technique does not discriminate between internal TT and internal rotation of the tibia. Thus, dogs with normal tibial conformation can be depicted by radiography as torsed, whereas dogs with TT may be misinterpreted as normal because of arbitrary positioning. CLINICAL RELEVANCE: Lateral displacement of the medial border of the calcaneus on a caudo-cranial radiograph should not be used as the sole arbiter of TT before surgical correction. PMID- 16266338 TI - Measurement of angles of abduction for diagnosis of shoulder instability in dogs using goniometry and digital image analysis. AB - OBJECTIVE: To compare abduction angles of shoulders with medial instability and unaffected shoulders in the same dogs and in age- and breed-matched dogs. STUDY DESIGN: Case-control study. ANIMALS: Dogs with medial instability of the shoulder (n=33) and 26 control dogs. METHODS: Dogs were sedated and positioned in lateral recumbency with both scapulas parallel to the table. With the elbow and shoulder in extension, the non-recumbent limb was maximally abducted and the angle between the scapular spine and lateral aspect of the brachium measured with a goniometer; a digital image was taken from the cranial aspect. Both techniques were performed in triplicate by 2 examiners. Mean abduction angles for each shoulder were determined from goniometric measurements and image analysis. Data were analyzed for significant differences between affected and unaffected shoulders, measurement techniques, and examiners. Strength of correlation between measurement techniques was determined. RESULTS: Mean abduction angles for shoulders with instability (53.7+/-4.7 degrees goniometric, 51.2+/-4.9 degrees image) were significantly (P<.001) larger than for all unaffected shoulders (32.6+/-2.0 degrees goniometric, 30.9+/-2.3 degrees image). In dogs diagnosed with instability, affected shoulders had significantly (P<.001) larger abduction angles than the contralateral (unaffected) shoulders. No significant differences were identified between right and left shoulders for control dogs, measurement techniques, or examiners. A strong (r=0.90) significant (P<.001) positive correlation between measurement techniques was noted. CONCLUSIONS: Shoulder abduction angles measured under sedation provide objective data for diagnosis of shoulder instability in dogs. Shoulders with clinical and arthroscopic evidence of medial instability have significantly higher abduction angles than shoulders that are considered normal. CLINICAL RELEVANCE: Determination of shoulder abduction angles should be included in the diagnostic protocol for forelimb lameness assessment in dogs. PMID- 16266339 TI - Treatment of dogs diagnosed with medial shoulder instability using radiofrequency induced thermal capsulorrhaphy. AB - OBJECTIVE: To report clinical findings and outcome in dogs diagnosed with medial shoulder instability (MSI) treated with radiofrequency-induced thermal capsulorrhaphy (RITC). STUDY DESIGN: Retrospective study. ANIMALS: Dogs (n=43) with MSI. METHODS: Medical records from consecutive cases with MSI based on orthopedic examination, palpation of shoulder abduction angles, and arthroscopic findings were used to determine clinical findings and long-term (> or = 1 year) outcome of RITC. RESULTS: Forty-three dogs met the criteria for inclusion; 2 dogs had bilateral MSI. The most common clinical findings were chronic unilateral weight-bearing lameness, large (> 45 degrees) shoulder abduction angles, and spinatus muscle atrophy in the affected limb. Mean preoperative abduction angle (50.7 degrees +/- 4.8 degrees) in affected shoulders was significantly larger than in the unaffected shoulders (32.5 degrees +/- 2.1 degrees). Mean postoperative abduction angle in affected shoulders was not significantly different from unaffected shoulders when measured immediately, 8, 16-20 weeks, and approximately 1 year after surgery. Follow-up evaluation ranged from 1 to 6.2 years. RITC treatment resulted in improved clinical function in 40 cases (93%) with 34 cases (79%) considered excellent; 3 cases (7%) were considered failures. CONCLUSIONS: RITC was a safe and effective method of treatment of MSI in most dogs studied. CLINICAL RELEVANCE: RITC can be considered as a viable option for surgical treatment of selected cases of MSI in dogs. PMID- 16266340 TI - Expression of relaxin receptor LRG7, canine relaxin, and relaxin-like factor in the pelvic diaphragm musculature of dogs with and without perineal hernia. AB - OBJECTIVES: To compare the expression of canine relaxin, relaxin-like factor (RLF), and relaxin receptors within the muscles of the pelvic diaphragm of dogs with perineal hernia (PH) and clinically normal dogs. STUDY DESIGN: In vivo comparative study. ANIMALS: Fifteen client-owned intact male dogs with PH were studied. Four mature intact male dogs with no evidence of perineal pathology served as controls. METHODS: Biopsy samples from the levator ani, coccygeus, and internal obturator muscles were obtained. RNA samples were reverse transcribed and analyzed by real-time PCR for the expression of canine relaxin receptor LRG7, relaxin, and RLF. RESULTS: Significantly higher expression levels of canine relaxin receptors occurred in the musculature of the pelvic diaphragm and internal obturator muscle in dogs with PH compared with normal dogs. Expression of canine RLF revealed no significant difference between dogs with PH and controls. The difference in the expression of canine relaxin between groups was not statistically significant. CONCLUSIONS: Relaxin receptor up-regulation occurs in the coccygeus, levator ani, and internal obturator muscles of dogs with PH. CLINICAL RELEVANCE: The higher expression of relaxin receptors within the muscles of the pelvic diaphragm in dogs with PH suggests that relaxin might play a role in the pathogenesis of PH. Atrophy of these muscles, which predisposes to PH, may be attributable to increased relaxin activity. PMID- 16266341 TI - Collagenolytic protease expression in cranial cruciate ligament and stifle synovial fluid in dogs with cranial cruciate ligament rupture. AB - OBJECTIVE: To determine expression of collagenolytic genes and collagen degradation in stifle tissues of dogs with ruptured cranial cruciate ligament (CCL). ANIMALS: Six dogs with CCL rupture and 11 dogs with intact CCL. PROCEDURES: Gene expression in CCL tissue and synovial fluid cells was studied using reverse transcriptase-polymerase chain reaction (RT-PCR). Collagen degradation was studied using CCL explant cultures and a synovial fluid bioassay. RESULTS: Expression of matrix metalloproteases (MMP) was not found in young Beagles with intact CCL; however, increased expression of MMP-3 was found in CCL tissue from older hounds with intact CCL, when compared with young Beagles. In dogs with ruptured CCL, expression of MMP-2 and -9 was increased in stifle tissues, when compared with dogs with intact CCL. Similar to MMP-9, expression of tartrate-resistant acid phosphatase (TRAP) and cathepsin S was only found in stifle tissues from dogs with ruptured CCL; in contrast, expression of cathepsin K was found in all ruptured and intact CCL. Collagen degradation was increased in ruptured CCL, when compared with intact CCL. CONCLUSION: Rupture of the CCL is associated with up-regulation of expression of MMP-2 and -9 (gelatinase A and B), TRAP, and cathepsin S, and increased degradation of collagen. CLINICAL RELEVANCE: These findings suggest that MMP-2, -9, cathepsin S, and TRAP may be important mediators of progressive joint destruction in dogs with CCL rupture. These genes are markers for macrophages and dendritic cells. MMP and cathepsin S pathways may offer novel targets for anti-inflammatory medical therapy aimed at ameliorating joint degradation associated with inflammatory arthritis. PMID- 16266342 TI - Biomechanical comparison of orthofix pins and cortical bone screws in a canine humeral condylar fracture model. AB - OBJECTIVE: To compare shear stability of simulated humeral lateral condylar fractures reduced with either a self-compressing pin or cortical bone screw. STUDY DESIGN: In vitro biomechanical tests. SAMPLE POPULATION: Bilateral cadaveric canine humeri (n=18) without evidence of elbow disease. METHODS: Lateral condylar fracture was simulated by standardized osteotomy. Bone fragments were stabilized with a self-compressing pin or a cortical bone screw (2.7 or 3.5 mm) inserted in lag fashion. Specimens were mounted in a materials testing system and the condylar fragment displaced in a proximal direction until failure. Mechanical testing variables derived from load-deformation curves were compared between stabilization methods using a Student's paired t-test. RESULTS: There were no statistically significant differences for mechanical testing variables between pin and screw stabilized specimens at expected walk and trot loads. Three yield points subjectively coincided with yield of the interfragmentary interface (Y1), bone at the implant interface (Y2), and implant deformation (Y3). Displacements at Y1 were 48-156% greater for pin than screw stabilized specimens. Y2 and Y3 loads were higher for screw than pin stabilized specimens, but likely supraphysiologic for dogs convalescing after surgical repair. CONCLUSIONS: A self compressing pin or a cortical bone screw inserted in lag fashion both provided adequate strength in applied shear to sustain expected physiologic loads through the repaired canine elbow during postoperative convalescence. CLINICAL RELEVANCE: Because self-compressing pins were easy to implant and mechanical properties were not significantly different than cortical screws at expected physiologic loads, pins should be considered for the repair of traumatic humeral condylar fractures. PMID- 16266343 TI - In vitro evaluation of screws and suture anchors in metaphyseal bone of the canine tibia. AB - OBJECTIVE: To compare ease of insertion, load to failure, and mode of failure of cortical and cancellous screws, BoneBiter, IMEX, and TwinFix suture anchors in canine metaphyseal tibial bone. STUDY DESIGN: Experimental biomechanical study. ANIMALS: Canine cadaveric tibias. METHODS: One investigator inserted all anchors and subjectively evaluated ease of placement. Anchor systems were loaded to failure along axis of insertion with audio-video recording to determine failure mode. RESULTS: BoneBiter was the most difficult anchor to insert successfully. Mean+/-SD loads to failure were cancellous screw (711+/-193 N), IMEX 4.7 mm 18 g wire (661+/-163 N), IMEX 4.0 mm 18 g wire (661+/-165 N), cortical screw (635+/ 184 N), BoneBiter #5 Kevlar suture (393+/- 109 N), and TwinFix 5.0 mm #2 polyester (267+/-73 N). No significant differences were noted among the cortical screw, cancellous screw, IMEX 4.7 and 4.0 mm, all of which were significantly (P<.001) greater than BoneBiter and TwinFix . Failure modes were pullout of bone, suture-wire breakage, eyelet breakage, or no failure to 1000 N: screws (18,0,0,2), IMEX (18,1,1,0), BoneBiter (2,8,0,0), and TwinFix (0,10,0,0). CONCLUSIONS: Fixation devices were user friendly, with the exception of BoneBiter. Mode of failure is dependent on suture material and anchor design. Cortical and cancellous screws, and IMEX anchors with 18 g wire have significantly greater load to failure compared with BoneBiter and TwinFix suture anchors. CLINICAL RELEVANCE: Based on load to failure, ease of use, design characteristics, and cost, IMEX anchors may have advantages over other comparable soft tissue fixation devices. PMID- 16266344 TI - Axial pattern flap based on the lateral caudal arteries of the tail in the dog: an experimental study. AB - OBJECTIVE: To determine applicability and size limits of an axial pattern flap based on the lateral caudal arteries in dogs to reconstruct caudodorsal trunk skin defects. STUDY DESIGN: Experimental study. ANIMALS: Ten mature, mixed breed dogs. METHODS: The lateral caudal vessels of the tail were incorporated in the flaps of the treatment group (n=5) and were ligated in the control group (n=5). Flaps were rotated and placed into experimentally created caudodorsal skin defects on the trunk. The length and area of tissue that remained viable in each flap were determined. RESULTS: Mean (+/-SD) survival area (222.8+/-32.9 cm2) and length (25.1+/-4.5 cm) of vascularized flaps were significantly greater (P<.05) compared with control flaps (94.9+/-13.4 cm2 and 14.61+/-4.7 cm). Necrosis occurred in all control flaps, resulting in lower percentage flap area (48.8%) and length (47.1%) survival compared with vascularized flaps (78.1%, 77.5%). CONCLUSIONS: Axial pattern flap based on lateral caudal arteries of the tail was successfully elevated and transferred in a single procedure, with 78% survival for closure of large experimentally created caudodorsal trunk defects in dogs. CLINICAL RELEVANCE: Large cutaneous defects of the caudodorsal trunk area in dog could be reconstructed with tail axial pattern flap. Limitations in terms of size and changes in animal appearance have to be considered before flap elevation. PMID- 16266345 TI - Repair of urethral defects using fascia lata autografts in dogs. AB - OBJECTIVE: To evaluate the feasibility of urethroplasty using a free fascia lata (FL) graft in the dog. STUDY DESIGN: In vivo experimental study. ANIMALS: Mixed breed dogs (n=14). METHODS: Half of the circumference of the urethra, approximately 1.5 cm long, was excised in 14 male dogs to induce a urethral defect. FL (approximately 2 cm x 2 cm) harvested from the lateral thigh was sutured to the urethra using a 3-0 polyglactin 910 continuous pattern. Dogs were monitored daily for bladder distention and had urethral catheters until normal voiding was observed. On day 60, each dog had a positive contrast urethrogram, and then 8 dogs were euthanatized for gross and histologic examination. Six dogs were monitored for urologic problems for 6 months, and a positive contrast urethrogram was repeated. RESULTS: All dogs recovered successfully; 4 dogs had difficulty voiding for 2-3 days and urine was aspirated from these dogs every 3 hours until signs of painful urination disappeared. On positive contrast urethrograms, urethral anatomy was considered normal except in 4 dogs that had an irregular contour. Gross urethral examination confirmed an absence of ulceration, stricture, diverticula, or fistula formation, and the FL-lined graft survived in all dogs. No degenerative and reparative responses were observed. On histologic examination of the penile urethra, the lumen was intact, covered with transitional epithelium, and surrounded by corpus spongiosum with cavernous spaces and blood-filled vessels. CONCLUSIONS: Free FL grafts are incorporated satisfactorily and would appear to be useful for repairing urethral defects. CLINICAL RELEVANCE: FL grafts should be considered for repair of urethral defects in dogs. PMID- 16266346 TI - Cisterna chyli ablation with thoracic duct ligation for chylothorax: results in eight dogs. AB - OBJECTIVE: To report use of combined cisterna chyli ablation (CCA) and thoracic duct ligation (TDL) for treatment of spontaneously occurring chylothorax in dogs. STUDY DESIGN: Retrospective study. ANIMALS: Eight dogs with chylothorax. METHODS: TDL was performed through a right caudal intercostal thoracotomy and CCA through a left flank paracostal approach or ventral median celiotomy. Long-term outcome (range, 2-48 months; median, 11.5 months) was evaluated by telephone communication with owners. RESULTS: Seven dogs were free of clinical signs related to chylothorax at last follow-up (range, 4-48 months; median, 15.5 months). One dog was euthanatized 2 months after surgery because of lack of improvement. No major complications occurred from CCA. CONCLUSION: CCA and TDL resolved chylothorax in most dogs (88%). CLINICAL RELEVANCE: CCA combined with TDL may improve the outcome of chylothorax in dogs. PMID- 16266347 TI - Trichophytobezoar duodenal obstruction in New World camelids. AB - OBJECTIVE: To describe clinical findings, surgical treatment, and outcome associated with trichophytobezoar duodenal obstruction in New World camelids. STUDY DESIGN: Retrospective study. ANIMALS: Alpacas (7) and 1 llama. METHODS: Historical and clinical data were obtained from the medical records of New World camelids with a diagnosis of trichophytobezoar duodenal obstruction confirmed by surgical exploration or necropsy. RESULTS: Seven camelids were <1 year old. Abnormal clinical findings included anorexia, reduced fecal output, recumbency, colic, abdominal distension, regurgitation, decreased serum chloride concentration, increased serum bicarbonate concentration, and/or elevated first gastric compartment chloride concentration. Survey abdominal radiographs obtained (4 animals) revealed gastric distension (4) and/or visualization of the obstruction (2). Diagnosis was confirmed at necropsy (1) or surgery (7). Right paracostal celiotomy was performed on all animals and duodenotomy (3) or retropulsion of the trichophytobezoar combined with third compartment gastrotomy (4) was used to remove the obstruction. Six animals survived to discharge and 5 were healthy at follow-up, 8-20 months later. The remaining discharged alpaca was healthy at 12 months but subsequently died of unrelated causes. CONCLUSIONS: Diagnosis of trichophytobezoar duodenal obstruction should be considered in juvenile New World camelids with abdominal distension and hypochloremic metabolic alkalosis. Right paracostal celiotomy can be used for access to the descending duodenum and third gastric compartment for surgical relief of obstruction. CLINICAL RELEVANCE: Duodenal obstruction from bezoars should be considered in New World camelids <1year of age with abdominal distension and hypochloremic metabolic alkalosis. Surgical relief of the obstruction by right paracostal celiotomy has a good prognosis. PMID- 16266348 TI - Thoracoscopic lung lobectomy for treatment of lung tumors in dogs. AB - OBJECTIVE: To report use of thoracoscopic lung lobectomy (TLL) for treatment of lung tumors (LT) in dogs. STUDY DESIGN: Retrospective study. ANIMALS: Nine dogs. METHODS: Dogs that had TLL for tumor removal were included. Using general anesthesia and 1-lung ventilation, TLL was performed using a 30-60 mm endoscopic gastrointestinal anastomosis stapler. If the visual field was obscured, lobe resection was completed via thoracotomy. RESULTS: Metastatic and primary LT were resected by thoracoscopic lobectomy in 9 dogs (6 male, 3 female; mean (+/-SD) weight, 29+/-7 kg; mean age, 10.7+/-1.9 years). Six dogs had a solitary mass and 3 dogs had 2 masses within a single lobe. The left caudal lobe was removed in 3 dogs. In 5 dogs, TLL was used alone whereas conversion to thoracotomy was required in 4 dogs because of poor visibility. There were 7 metastatic LT and 2 primary LT. Mean duration of thoracoscopic surgery was 108.8+/-30.3 minutes compared with 150.75+/-55.4 minutes in dogs requiring conversion to thoracotomy. Mean hospitalization was 3.1+/-1.3 days. CONCLUSION: Provided the visual field is not obscured, TLL can be performed effectively in dogs. CLINICAL RELEVANCE: Dogs with metastatic or primary LTs should be considered for TLL, particularly for small masses positioned away from the hilus in the left caudal lung lobe. PMID- 16266350 TI - Evidence-based medicine and cranial cruciate ligament repairs. PMID- 16266352 TI - Do patient expectations and diagnostic specificity affect outcomes in pharmacological trials in pain medicine. PMID- 16266351 TI - Chest tube drainage circuitry in mammals. PMID- 16266354 TI - Multidimensional success criteria and expectations for treatment of chronic pain: the patient perspective. AB - OBJECTIVE: The study's primary objective is to define treatment success from the patient perspective across four domains (pain, fatigue, emotional distress, interference with daily activities) using the Patient Centered Outcomes Questionnaire (PCOQ). Secondary objectives include examination of patient expectations for treatment across domains and derivation of patient subgroups based on PCOQ responses. DESIGN: Cross-sectional. Setting. University of Florida or affiliated pain clinics. Patients. Participants were 110 patients seeking treatment for chronic pain who completed the PCOQ before their routine medical appointment. RESULTS: Patients reported moderate usual levels of pain (mean = 6.1), fatigue (mean = 5.9), distress (mean = 5.5), and interference (mean = 6.4). Patients considered a mean reduction in pain of 3.4 points (56%) to represent treatment success. Mean reductions in fatigue, distress, and interference of 3.35 points (57%), 3.64 points (65%), and 4.3 points (68%), respectively, also signified treatment success. Analyses revealed patients expected their level of interference following treatment (mean = 2.6) to be higher than their success criterion for interference (mean = 2.0). Cluster analysis of PCOQ responses revealed a three-cluster solution. Differences were obtained among clusters on demographic and pain-related variables. CONCLUSIONS: Patients might require larger reductions in pain than has previously been reported in the literature as "meaningful" for them to consider treatment successful. Patients did not expect treatment to meet their criterion for success in the interference domain, suggesting patients' success criteria and treatment expectations might differ for some domains. The finding of patient subgroups has implications for treatment matching. PMID- 16266355 TI - A double-blind, randomized multicenter trial comparing duloxetine with placebo in the management of diabetic peripheral neuropathic pain. AB - OBJECTIVE: Assess efficacy and safety of duloxetine, a selective serotonin and norepinephrine reuptake inhibitor, on the reduction of pain severity, in patients with diabetic peripheral neuropathic pain (DPNP). METHODS: This was a multicenter, parallel, double-blind, randomized, placebo-controlled trial that enrolled 348 patients with pain due to peripheral neuropathy caused by type 1 or type 2 diabetes mellitus. Patients (N = 116 per group) were randomly assigned to receive duloxetine 60 mg once daily (QD), duloxetine 60 mg twice daily (BID), or placebo, for 12 weeks. The primary outcome measure was the weekly mean score of 24-hour average pain severity evaluated on an 11-point Likert scale. Secondary outcome measures and safety were evaluated. RESULTS: Compared with placebo treated patients, both duloxetine-treated groups improved significantly more (P < 0.001) on the 24-hour average pain score. Duloxetine demonstrated superiority to placebo in all secondary analyses of the primary efficacy measure. A significant treatment effect for duloxetine was observed in most secondary measures for pain. Discontinuations due to adverse events were more frequent in the duloxetine 60 mg BID- (12.1%) versus the placebo- (2.6%) treated group. Duloxetine showed no adverse effects on diabetic control, and both doses were safely administered and well tolerated. CONCLUSIONS: In this clinical trial, duloxetine 60 mg QD and duloxetine 60 mg BID were effective and safe in the management of DPNP. PMID- 16266356 TI - Oxymorphone extended-release tablets relieve moderate to severe pain and improve physical function in osteoarthritis: results of a randomized, double-blind, placebo- and active-controlled phase III trial. AB - OBJECTIVE: To compare oxymorphone extended release (ER) and placebo on indices of pain, function, and safety in patients with chronic osteoarthritis (OA) pain. DESIGN: In this multicenter, double-blind, placebo- and active-controlled, parallel-group, dose-ranging study, patients were randomized to oxymorphone ER 20 mg (N = 121), oxymorphone ER 40 mg (N = 121), oxycodone controlled release 20 mg (N = 125), or placebo (N = 124) every 12 hours. The primary efficacy end point was change in arthritis pain intensity (visual analog scale, 0-100) from baseline to week 3 for the oxymorphone ER 40 mg group versus placebo. RESULTS: The primary end point was achieved: the week 3 oxymorphone ER least squares mean difference (LSMD) from placebo was -9.0 (95% confidence interval [CI]: -16.2 to -1.8; P = 0.015). Secondary efficacy analysis showed similar improvements at week 4 (LSMD from placebo, -10.3 [95% CI: -17.7 to -2.8]; P = 0.007) and with oxymorphone ER 20 mg at week 3 (LSMD from placebo, -7.7 [95% CI: -15.0 to -0.4]; P = 0.039) and week 4 (LSMD from placebo, -7.5 [95% CI: -15.0 to 0.0]; P = 0.050). Weeks 3 and 4 pain intensity decreased by approximately 30-40%. Oxymorphone ER 20 and 40 mg improved from baseline on the Western Ontario and McMaster Universities Osteoarthritis Composite Index and pain and physical function subscales at week 4. Adverse events in all opioid groups included mild to moderate nausea, constipation, and somnolence. CONCLUSIONS: In this short-term study, oxymorphone ER was superior to placebo for relieving pain and improving function in patients with moderate to severe chronic OA pain, and is an alternative to other sustained release opioids. PMID- 16266357 TI - Visual impairment following epidural fluid injections and epiduroscopy: a review. AB - INTRODUCTION: Visual impairment apparently is a rare but significant complication following epidural fluid injection and epiduroscopy. We report a review of cases reported in the literature with the aim of gaining an understanding of how the complication occurs as well as how to avoid it. MATERIALS AND METHODS: A literature review was performed using PubMed to find 1) case reports and 2) factors that contribute to the development of visual loss in the cases. RESULTS: Twelve cases of visual impairment following epiduroscopy or epidural fluid injection have been reported in the literature. The average age of the patients was approximately 50 years, 83.3% of the patients were female, 16.7% were male. Bolus injection of fluid with or without epiduroscopy was considered to be the precipitating event. The volume of solution injected epidurally varied from 20 mL to 120 mL (average = 59.4 mL). The common finding was retinal hemorrhage, with 58.3% occurring bilaterally. Recovery occurred in 79.2% of the cases. CONCLUSION: Retinal hemorrhage following epidural fluid injections/epiduroscopy apparently is due to an increase in cerebrospinal fluid pressure proportional to the rate and the amount of fluid injected. A sudden increase in epidural pressure is transmitted into the subarachnoid space to the optic nerve sheath, compressing the optic nerve and its vasculature. The vasculature compression ruptures retinal blood vessels. PMID- 16266358 TI - Topiramate for phantom limb pain: a time-series analysis. AB - There is growing evidence of topiramate's efficacy in treating neuropathic pain. This article reports a detailed analysis of the response of four amputee subjects with phantom limb pain. Individual time-series analyses revealed that three out of four amputee participants receiving topiramate had statistically significant decreases in pain, with the peak effect noted at 800 mg daily. This analysis supports a hypothesis that topiramate may be effective in reducing phantom limb pain, and suggests a definitive study is indicated. PMID- 16266359 TI - An open-label study of the lidocaine patch 5% in painful idiopathic sensory polyneuropathy. AB - OBJECTIVE: Painful idiopathic distal sensory polyneuropathy is common, but has been largely ignored as a model for the evaluation of neuropathic pain therapies. We have therefore conducted a safety, tolerability, and effectiveness study of the lidocaine patch 5% in painful idiopathic distal sensory polyneuropathy. DESIGN: A prospective open-label, flexible dosing, 3-week study period with a 5 week extension. SETTING: Peripheral Neuropathy clinics and Anesthesiology Clinical Research Center at a tertiary care facility. PATIENTS: Twenty subjects with a diagnosis of idiopathic distal sensory polyneuropathy (with or without associated impaired glucose tolerance), with a baseline mean pain daily rating of > or =4 on a visual analog scale. Intervention. Lidocaine patch 5%, maximum of four patches daily for 18 hours. MAIN OUTCOME MEASURE: Change from baseline week to week 3 mean daily diary pain ratings. Secondary endpoints included assessments of safety and tolerability as well as quality of life measures. RESULTS: Subjects with idiopathic distal sensory polyneuropathy, both with and without impaired glucose tolerance, showed significant improvements in pain and quality of life outcome measures over a 3-week treatment period. These improvements were maintained in a subgroup of patients treated for an additional 5 weeks and permitted a taper of concomitant analgesics in 25% of subjects. The lidocaine patch 5% was well tolerated. CONCLUSIONS: The lidocaine patch 5% appeared well tolerated and potentially effective in the management of painful idiopathic distal sensory polyneuropathy. Idiopathic distal sensory polyneuropathy is an appropriate patient population for the conduct of clinical trials of neuropathic pain therapies. PMID- 16266360 TI - Trustworthiness as a clinical variable: the problem of trust in the management of chronic, nonmalignant pain. AB - The subjective nature of pain leads to many treatment difficulties. These problems can often be resolved if we know that the patient is trustworthy. Trustworthiness should be assessed as a distinct clinical variable. This is more easily achieved if we examine the three components of trustworthiness: the patient's subjective reports, which we call testimony; the reason that the patient seeks treatment, which we call motive; and the patient's adherence with efforts to get well, which we call responsibility. Because of difficulties with assessing testimony and motive, we propose that establishing the patient's responsibility is the key to assessing trustworthiness. PMID- 16266361 TI - Trust and pharmaco-vigilance in pain medicine. PMID- 16266362 TI - The doctor as double agent. PMID- 16266366 TI - A mutation of ion-conducting pore without effect on ion selectivity of the sodium channel. PMID- 16266367 TI - Cardiac gene expression profiling may reveal key differences between physiologic and pathologic cardiac hypertrophy. PMID- 16266368 TI - Gene expression profiling of exercise-induced cardiac hypertrophy in rats. AB - AIMS: Exercise training causes physiological cardiac hypertrophy, which acts to enhance cardiac function during exercise. However, the underlying molecular mechanisms are unclear. We investigated gene expression profile of exercise training-induced cardiac hypertrophy using left ventricle (LV) excised from exercise-trained and sedentary control rats (12-week old). METHOD: Rats in the training group exercised on a treadmill for 8-week. RESULTS: Left ventricular mass index and wall thickness in the exercise-trained group were significantly greater than that in the control group, indicating that the trained rats developed cardiac hypertrophy. Of the 3800 genes analysed in the microarray analyses, a total of 75 relevant genes (upregulation of 33 genes and downregulation of 42 genes) displayed alterations with exercise training. Among these genes, we focused on glycogen synthase kinase (GSK)-3beta, calcineurin inhibitor (Cain), and endothelin (ET)-1 for their implicated roles in pathological cardiac hypertrophy, and confirmed the results of microarray analysis at mRNA and protein/peptide levels using quantitative PCR, Western blot, and EIA analyses. The gene expression of GSK-3beta decreased significantly and those of Cain and ET-1 increased significantly with exercise training. Furthermore, LV mass index was significantly correlated with GSK-3beta protein activity (r = -0.70, P < 0.01) and tissue ET-1 concentration (r = 0.52, P < 0.05). There were no changes in gene expressions in brain natriuretic peptide (BNP), angiotensin-correcting enzyme (ACE), interleukin-6, and vascular cell adhesion molecule (VCAM)-1. CONCLUSION: These findings suggest that physiological and pathological LV hypertrophy may share some of the same molecular mechanisms in inducing LV hypertrophy (e.g. GSK-3beta, Cain, and ET-1) and that other genes (e.g. BNP, ACE) may differentiate physiological from pathological LV hypertrophy. PMID- 16266369 TI - Ion channel regulation of the dynamical instability of the resting membrane potential in saccular hair cells of the green frog (Rana esculenta). AB - AIMS: We investigated the ion channel regulation of the resting membrane potential of hair cells with the aim to determine if the resting membrane potential is poised close to instability and thereby a potential cause of the spontaneous afferent spike activity. METHODS: The ionic mechanism and the dynamic properties of the resting membrane potential were examined with the whole-cell patch clamp technique in dissociated saccular hair cells and in a mathematical model including all identified ion channels. RESULTS: In hair cells showing I/V curves with a low membrane conductance flanked by large inward and outward rectifying potassium conductances, the inward rectifier (K(IR)), the delayed outward rectifier (K(V)) and the large conductance, calcium-sensitive, voltage gated potassium channel (BK(Ca)) were all activated at rest. Under current clamp conditions, the outward current through these channels balanced the inward current through mechano-electrical transduction (MET) and Ca2+ channels. In 45% (22/49) of the cells, the membrane potential fluctuated spontaneously between two voltage levels determined by the voltage extent of the low membrane conductance range. These fluctuations were not influenced by blocking the MET channels but could be reversibly stopped by increasing [K+]o or by blocking of K(IR) channels. Blocking the BK(Ca) channels induced regular voltage oscillations. CONCLUSIONS: Two intrinsic dynamical instabilities of V(m) are present in hair cells. One of these is observed as spontaneous voltage fluctuations by currents through K(IR), K(V) and h-channels in combination with a steady current through MET channels. The other instability shows as regenerative voltage changes involving Ca2+ and K(V) channels. The BK(Ca) channels prevent the spontaneous voltage fluctuations from activating the regenerative system. PMID- 16266370 TI - Novel Brugada syndrome-causing mutation in ion-conducting pore of cardiac Na+ channel does not affect ion selectivity properties. AB - AIM: Brugada syndrome is an inherited cardiac disease with an increased risk of sudden cardiac death. Thus far Brugada syndrome has been linked only to mutations in SCN5A, the gene encoding the alpha-subunit of cardiac Na+ channel. In this study, a novel SCN5A gene mutation (D1714G) is reported, which has been found in a 57-year-old male patient. Since the mutation is located in a segment of the ion conducting pore of the cardiac Na+ channel, which putatively determines ion selectivity, it may affect ion selectivity properties. METHODS: HEK-293 cells were transfected with wild-type (WT) or D1714G alpha-subunit and beta-subunit cDNA. Whole-cell configuration of the patch-clamp technique was used to study biophysical properties at room temperature (21 degrees C) and physiological temperature (36 degrees C). This study represents the first measurements of human Na+ channel kinetics at 36 degrees C. Ion selectivity, current density, and gating properties of WT and D1714G channel were studied. RESULTS: D1714G channel yielded nearly 80% reduction of Na+ current density at 21 and 36 degrees C. At both temperatures, no significant changes were observed in V(1/2) values and slope factors for voltage-dependent activation and inactivation. At 36 degrees C, but not at 21 degrees C, D1714G channel exhibited more slow inactivation compared with WT channel. Ion selectivity properties were not affected by the mutation at both temperatures, as assessed by either current or permeability ratio. CONCLUSION: This study shows no changes in ion selectivity properties of D1714G channel. However, the profoundly decreased current density associated with the D1714G mutation may explain the Brugada syndrome phenotype in our patient. PMID- 16266372 TI - A single bout of exercise is followed by a prolonged decrease in the interstitial glucose concentration in skeletal muscle. AB - AIM: The present study was performed to test the hypothesis that the interstitial glucose concentration in human skeletal muscle is decreased for a prolonged period following a single bout of exercise, while blood flow has returned to resting levels. METHODS: Muscle interstitial concentrations of glucose, lactate, pyruvate and urea were monitored in six healthy individuals during 8 h following a 2-h one-leg exercise session by microdialysis at low perfusion flow rate. Simultaneously the blood flow was measured by the microdialysis ethanol technique. RESULTS: The blood glucose and the control leg interstitial glucose concentrations were stable during the experiment averaging 5.7 +/- 0.1 and 4.1 +/ 0.3 mm, respectively. In contrast, the interstitial glucose concentration in the exercise leg was markedly decreased, averaging 1.9 +/- 0.5 mm, during the first 5.5 h following exercise (P < 0.01), after which it returned towards normal values. Muscle blood flow at the site of the microdialysis catheter, measured as the ethanol outflow-to-inflow ratio, did not change significantly over time in the control or exercise leg and did not differ significantly between the two legs. Interstitial concentrations of lactate, pyruvate and urea were not significantly different between the control and exercise leg. CONCLUSION: The study shows that the interstitial glucose concentration in skeletal muscle is markedly decreased for several hours following a single exercise session. The decreased interstitial glucose concentration may serve to limit the rate of post exercise muscle glucose uptake to a rate compatible with normal blood glucose levels and may also be speculated to have a positive long-term health implication by augmenting muscle insulin sensitivity. PMID- 16266371 TI - Escherichia coli derived factors modulate human granulosa cell steroidogenesis. AB - AIM: In order to identify bacterial compounds which directly affect granulosa cell steroidogenesis, the effect of culture-supernatants and extracts from Escherichia coli were tested in an in vitro granulosa cell culture model. METHODS: Samples were drawn from the culture-supernatant of E. coli ATCC25922 cultured in Luria-broth medium. Bacterial extract was prepared by incubation of freshly harvested bacteria in buffer. The bacterial culture-supernatants and extracts were added to human granulosa cells in culture. The granulosa cells collected from the follicular aspirates from women undergoing in-vitro fertilization were cultured for 17-53 h and progesterone or oestradiol was assayed in the spent culture medium. RESULTS: The E. coli culture-supernatant stimulated the basal granulosa cell progesterone production demonstrating its maximum activity reached after 200-240 min of bacterial growth. The heat denaturated bacterial extract as well as its low-molecular-weight fraction (<10 kDa) stimulated both the basal and the hCG-stimulated progesterone production; the oestradiol production was slightly inhibited. Stimulation of progesterone production was time dependent increasing from 125 +/- 18% of control within the first 3 h to 205 +/- 35% within 17-53 h. The high-molecular-weight fraction (>30 kDa) of the bacterial extract inhibited progesterone production. The inhibitory activity was significantly diminished by heat denaturation. CONCLUSIONS: The present study demonstrates the existence of various compounds which are secreted by E. coli and could also be extracted out of E. coli bacteria. These bacterial compounds modulate ovarian steroidogenesis. Further studies are needed to clarify how far these compounds contribute to menstrual disturbance observed in chronic pelvic inflammation. PMID- 16266373 TI - Neuromuscular function during prolonged pedalling exercise at different cadences. AB - AIM: The purpose of the present work was to assess the strategies set by the central nervous system in order to provide the power output required throughout a prolonged (1-h) pedalling exercise performed at different cadences (50 rpm, 110 rpm and the freely chosen cadence). METHODS: Neuromuscular (NM) activity of vastus lateralis, rectus femoris, biceps femoris and gastrocnemius lateralis muscles was studied quantitatively [root-mean square (RMS) and mean power frequency (MPF)] and qualitatively (timing of onset and offset of muscle bursts during crank cycle). RESULTS: The present results showed that increased cadence resulted in earlier muscle activation in crank cycle. The influence of cadence on RMS and MPF depended on the considered muscle and its functional role during pedalling. Timing of onset and offset of muscle bursts was not altered by fatigue throughout the prolonged exercise. In contrast, RMS and MPF of some muscles was found to increase during prolonged exercise. CONCLUSION: In summary, the present study revealed that tonic aspects of the NM activity (RMS, MPF) are altered during prolonged pedalling exercise, while phasic aspects are remained unchanged. These results suggest that the strategies set by the central nervous system in order to provide the power output required by the exercise are held constant throughout the exercise, but that quantitative aspects of the central drive are increased in order to adapt to the progressive occurrence of the NM fatigue. PMID- 16266374 TI - Protein kinases A and C stimulate the Na+ active transport in frog skeletal muscle without an appreciable change in the number of sarcolemmal Na+ pumps. AB - AIM: The activation of both protein kinases A (PKA) and protein kinases C (PKC) in some cell types increases and in others reduces active Na+ efflux. These effects have been ascribed to either a change in the rate of ionic translocation by a fixed number of Na+ pumps or, a change in the number of plasma membrane pumps. The purpose of the present experiments was to study the effect of activating PKA and PKC on the Na+ extrusion by the Na+ pump in frog skeletal muscle. METHODS: Na+ (22Na+) fluxes and ouabain (3H-ouabain) binding were measured in frog sartorius muscles. RESULTS: Both activation of PKA and PKC increased the active Na+ extrusion by a factor of two; these effects were not additive. Ouabain binding experiments indicated that the pump stimulation by activation of these kinases is not associated with any significant increase in the number of plasma membrane pumps. Stimulation of the active Na+ efflux by protein kinase activation (no change in the number of sarcolemmal pumps) and by hypotonicity (increase in the number of pumps) could be elicited in the same preparation and they were additive. CONCLUSION: It is concluded that in frog skeletal muscle fibres, (1) activation of both PKA and PKC stimulate the Na+ pump by increasing its rate of ionic translocation; and (2) two modes of Na+ active transport (with and without an increase in the number of pumps) are operative, and can be at work simultaneously, a phenomenon to be reckoned with. PMID- 16266375 TI - Comparison on plasma caesium kinetics in goats and horses with special emphasis on exercising horses. AB - AIMS: Like potassium (K+), caesium (Cs+) tends to concentrate intracellularly. The aim here was to determine how moderate exercise affects the uptake of Cs+ from blood plasma. METHODS: After an intravenous Cs+ dose of 5 micromol kg(-1), plasma Cs+ concentration was followed for 100 min in goats and for 60 min in horses. The latter were divided into two groups, one resting and the other trotting on a treadmill (inclination 3 degrees, speed 5 m s(-1)). RESULTS: The plasma Cs+ concentration follows a multiphase exponential decay curve, which initially could be approximated with a two-phase curve. The initially high rate constant (approximately 10 h(-1)) decreased to around 1 h(-1) in 40 min. Exercise more than doubled the rate of removal of Cs+ from plasma between 20 and 40 min after the start of exercise. After exercise, the rate returned to resting levels within 10 min. Plasma K+, on the contrary, declined for at least 20 min after exercise had ended. CONCLUSIONS: Moderate exercise significantly increases the rate of removal of Cs+ from the bloodstream. After exercise, the rate returns to the resting levels within 10 min. The increased rate of Cs+ removal during exercise is likely due to increased activity of Na+, K+-ATPase in working skeletal muscles. PMID- 16266377 TI - Idiopathic rhinitis, the ongoing quest. AB - The term rhinitis in daily practice is used for nasal dysfunction causing symptoms-like nasal itching, sneezing, rhinorrhea and or nasal blockage. Chronic rhinitis can roughly be classified into allergic, infectious or nonallergic/noninfectious. When allergy, mechanical obstruction and infections have been excluded as the cause of rhinitis, a number of poorly defined nasal conditions of partly unknown aetiology and pathophysiology remain. The differential diagnosis of nonallergic noninfectious rhinitis is extensive. Although the percentage of patients with nonallergic noninfectious rhinitis with a known cause has increased the last decades, still about 50% of the patients with nonallergic noninfectious rhinitis has to be classified as suffering from idiopathic rhinitis (IR), or rather e causa ignota. Specific immunological, clinical and sometimes radiological and functional tests are required to distinguish known causes. Research to the underlying pathophysiology of IR has moved from autonomic neural dysbalans to inflammatory disorders (local allergy), the nonadrenergic noncholinergic (NANC) sensory peptidergic neural system and central neural hyperaesthesia, still without solid ground or proof. This review summarizes the currently known causes for nonallergic noninfectious rhinitis and possible treatments. Also possible pathophysiological mechanisms of IR are discussed. PMID- 16266376 TI - Prevention and treatment of hymenoptera venom allergy: guidelines for clinical practice. AB - Based on the knowledge of the living conditions and habitat of social Aculeatae a series of recommendations have been formulated which can potentially greatly minimize the risk of field re-sting. After a systemic sting reaction, patients should be referred to an allergy specialist for evaluation of their allergy, and if necessary venom immunotherapy (VIT). An emergency medical kit should be supplied, its use clearly demonstrated and repeatedly practised until perfected. This should be done under the supervision of a doctor or a trained nurse. Epinephrine by intramuscular injection is regarded as the treatment of choice for acute anaphylaxis. H1-antihistamines alone or in combination with corticosteroids may be effective in mild to moderate reactions confined to the skin and may support the value of treatment with epinephrine in full-blown anaphylaxis. Up to 75% of the patients with a history of systemic anaphylactic sting reaction develop systemic symptoms once again when re-stung. Venom immunotherapy is a highly effective treatment for individuals with a history of systemic reaction and who have specific IgE to venom allergens. The efficacy of VIT in yellow jacket venom allergic patients has been demonstrated also by assessing health related quality of life. If both skin tests and serum venom specific IgE turn negative, VIT may be stopped after 3 years. After VIT lasting 3-5 years, most patients with mild to moderate anaphylactic symptoms remain protected following discontinuation of VIT even with positive skin tests. Longer term or lifelong treatment should be considered in high-risk patients. Because of the small but relevant risk of re-sting reactions, in these patients, emergency kits, including epinephrine auto-injectors, should be discussed with every patient when stopping VIT. PMID- 16266378 TI - Genetics in asthma: the solution to a lasting conundrum? PMID- 16266379 TI - The effect of polymorphisms at the CD14 promoter and the TLR4 gene on asthma phenotypes in Turkish children with asthma. AB - BACKGROUND: Endotoxin, with its potential to enhance type 1 immunity, is a significant player in the hygiene hypothesis. The combined effects of the genetic variants of various molecules in the endotoxin response pathway on asthma related phenotypes are largely unknown. OBJECTIVE: To investigate the effects of the genetic variants of CD14 and TLR4 genes on asthma phenotypes in a large number of asthmatic children. METHODS: 613 asthmatic children were genotyped at the CD14 C159T, TLR4-A896G and TLR4-C1196T loci. IgE, eosinophil numbers and FEV1 were compared in 327 children who were not on any controller medications and were symptom free. Multivariate logistic regression was used to determine the factors associated with total IgE. RESULTS: Among children with atopic asthma, total IgE levels were significantly different among the three genotypes in the co-dominant model [CC: 435 kU/l (interquartile range: 146-820); CT: 361 (140-710); TT 204 (98 435), P = 0.035]. TT genotype was significantly and independently associated with lower IgE levels (OR: 0.5 95%; CI = 0.28-0.90, P = 0.021). Both TLR4-A896G and TLR4-C1196T polymorphisms were more frequent in the mild asthma group with atopy (P = 0.032, 0.018, respectively). The combined effects of the genetic variants in CD14 and TLR4 genes did not improve the observed associations. CONCLUSION: Our study demonstrates that the CD14-C159T promoter variant influences total IgE levels and also indicates that the T allele has a more profound effect on total IgE in children with atopic asthma. Polymorphisms in the TLR4 gene may be associated with milder forms of disease in atopic asthmatics in the population studied. PMID- 16266380 TI - Exhaled nitric oxide and exercise-induced bronchoconstriction in young male conscripts: association only in atopics. AB - This study was conducted to evaluate how bronchial responsiveness to direct and indirect stimuli relate to nitric oxide producing airway inflammation, and whether the relationship differs between atopic and nonatopic patients with various degrees of bronchial hyperresponsiveness and airway inflammation in a group of otherwise homogenous young men. We studied 181 consecutive non-smoking steroid-naive young male conscripts referred to military hospital because of respiratory symptoms suggesting asthma. Skin prick tests, spirometry, measurement of exhaled nitric oxide (FENO), and standardized airway challenges with histamine and exercise were performed. 128 patients were atopic. FENO was significantly higher in the atopic group, median 21.2 ppb, compared to 10.2 ppb in the nonatopic group. Still, 36% of all nonatopic patients had elevated FENO. Bronchial responsiveness to histamine (HIB) was similar in the two groups, but exercise-induced bronchoconstriction (EIB) was stronger in atopics (P < 0.01). FENO associated significantly with atopy (P < 0.001), severity of EIB (P < 0.001) and HIB (P = 0.006) in multiple linear regression model. In separate regression models for atopic and nonatopic patients FENO associated with severity of EIB and HIB in atopic patients only. The results were similar when patients with confirmed diagnosis of asthma were analyzed separately. Our results indicate that FENO significantly associates with EIB and HIB in atopic, but not in nonatopic steroid-naive patients with asthmatic symptoms. The finding suggests that in such atopic patients degree of airway hyperresponsiveness may reflect severity of airway inflammation. However, in nonatopic patients with similar symptoms other mechanisms of airway hyperresponsiveness may be more important. PMID- 16266381 TI - Exhaled nitric oxide predicts airway hyper-responsiveness to hypertonic saline in children that wheeze. AB - BACKGROUND: Exhaled nitric oxide (eNO) has shown good validity for the assessment of airway inflammation in asthmatic children. In large-scale epidemiological studies, this method would be preferred above airway challenge tests, because it is a quick and easy applicable tool. OBJECTIVE: In this study, we aimed to assess the discriminatory capacity of eNO, and prechallenge FEV1 for airway hyper responsiveness (AHR) in 8-13-year old schoolchildren. MATERIALS AND METHODS: Parents completed the ISAAC questionnaire, and children were tested for atopy, AHR to hypertonic (4.5%) saline (HS), and eNO. Diagnostic value was assessed by the area under the receiver operating curves (ROC), and calculation of positive and negative predicted values at different cut-off points for eNO and prechallenge FEV1. RESULTS: Areas under the ROC-curves of AHR were 0.65 for eNO and 0.62 for FEV1. Values increased to 0.71 and respectively 0.75 for a combined occurrence of AHR and current wheeze. Highest sensitivity and specificity were obtained at a cut-off value of 43 ppb for eNO and 103% predicted for FEV1. At these cut-off values, the positive predictive values for the presence of AHR in symptomatic children were respectively 83% (eNO) and 33% (FEV1), and negative predictive values in asymptomatic children were, respectively, 90 (eNO) and 80% (FEV1). CONCLUSION: Exhaled nitric oxide is a valid screening tool for AHR to HS in children that present with current wheeze, and it outperforms FEV1 as a predictor of AHR. PMID- 16266382 TI - Quality of asthma care: results from a community pharmacy based survey. AB - BACKGROUND: Optimal control is a major objective of disease management of asthma. The aim of the present study was to provide descriptive data on disease management in asthma patients, including medical resource utilization. METHODS: Asthma patients (age 18-50 years) were consecutively recruited in 348 dispensing pharmacies. They completed a questionnaire which collected data on personal characteristics, asthma management, including medical resource utilization, including asthma management. Asthma control was measured with the Asthma Control Test. Data from computerized pharmacy records of medications, dispensed before inclusion, were also collected. RESULTS: In 1791 eligible patients, 1559 accepted to participate in the study (mean age = 36.5, 56.1% of females). During the previous 4 weeks, the asthma control was satisfactory for only 28% of the patients, despite extensive provision of anti-inflammatory asthma control treatments (89%). Combinations of long acting beta agonists (LABA) and inhaled corticosteroids (ICS) were commonly used (59%), while fewer patients received LABA and ICS as two separate medications (15%). In addition, short-acting beta agonists, were frequently dispensed (71%). A substantial number of patients consulted their GPs on a monthly basis. Patients commonly reported daily shortness of breath (30%), daily use of rescue medication (29%) and weekly nocturnal symptoms (32%). Surprisingly, most patients considered their asthma as completely or well controlled (76%). CONCLUSIONS: Our results clearly identify a need to improve the management of asthma. Education programmes would be beneficial to improve asthma control. PMID- 16266383 TI - Effects of low doses of inhaled fluticasone propionate on inflammation and remodelling in persistent-mild asthma. AB - In asthma a dysregulation of eosinophil apoptosis and an imbalance of metalloproteinase-9 (MMP-9) and tissue inhibitor metalloproteinase-1 (TIMP-1) play an important role in airway inflammation and remodelling. We evaluated the effects of a low dose of inhaled fluticasone proprionate (FP) (100 microg bid by Diskus) for 4 weeks in 24 steroid naive patients with mild persistent asthma, symptomatic and with a sputum eosinophilia >or=3% on clinical outcomes and inflammatory markers such as the induced sputum eosinophils, the induced sputum apoptotic eosinophils, the levels of MMP-9 and TIMP-1 and their molar ratio in the induced sputum supernatants. After FP treatment forced expiratory volume (FEV1) and FEV1/forced vital capacity values, PEF (L/min), sputum apoptotic eosinophils, and MMP-9/TIMP-1 molar ratio in sputum supernatants of asthmatic subjects were significantly increased in comparison with baseline, while sputum eosinophils significantly decreased. Change (Delta) in FEV1 after treatment with FP negatively correlated with the Delta in sputum eosinophils, while the Delta in MMP-9 values positively correlated with Delta in TIMP-1 values. This study shows that the clinical improvement achieved by the use of low doses of FP in asthmatics is related, at least in part, to the resolution of eosinophilic inflammation and the downregulation of remodelling markers. PMID- 16266384 TI - Hypersensitivity to Forcipomyia taiwana (biting midge): clinical analysis and identification of major For t 1, For t 2 and For t 3 allergens. AB - BACKGROUND: Forcipomyia taiwana is a tiny, blood-sucking midge that cause intense pruritus and swelling in sensitive individuals. It is distributed island-wide in rural Taiwan and Southern China. OBJECTIVE: This study aimed to study the allergic immune responses and identify F. taiwana allergens. METHODS: Crude whole body F. taiwana extracts were prepared with phosphate-buffered saline. The specific IgE antibody was determined by enzyme-linked immunoassay and immunoblotting. Protein was analyzed by electrospray ionization tandem mass spectrometry. RESULTS: Among the 372 subjects that were exposed to F. taiwana bites, 179 (48%) reported an immediate skin reaction with/without delay reaction and 41(11.1%) reported a solely delay reaction. The skin of 21 subjects was tested with F. taiwana extract. Of these 21 subjects, 12 (57.1%) produced immediate skin reactions and contained high levels of specific IgE antibody against F. taiwana. Immunoblotting revealed that 11 allergenic components are able to bind specific IgE. Allergens of 22, 24, 35, 36, and 64 kDa bound 50, 50, 75, 66.7, and 75% of IgE-containing sera tested, respectively. Tryptic fragments of the 24, 35, 36, and 64 kDa allergens were analyzed by ESI-MS/MS. Selected tryptic peptides of 24, 35, and 36, and 64 kDa allergens exhibited significant sequence identity with triosephosphate isomerase of Anopheles merus,Tenebrio molitor,Ochlerotatus togoi, and Chrysops vittatus, fructose 1,6-bisphosphate aldolase of Antheraea yamamai and Homalodisca coagulata, and a slow muscle myosin S1 heavy chain of Homarusamericanus and a protein with unknown function from A. gambiae, respectively. The 35 and 36 kDa proteins may represent different isoforms of the fructose 1,6-bisphosphate aldolase. CONCLUSION: We conclude that immediate reaction to F. taiwana bites is IgE mediated and the 24 (For t 1), 35 (For t 2), and 64 kDa (For t 3) proteins are candidates for major F. taiwana allergens. Further studies are needed to confirm these allergens. PMID- 16266385 TI - Single dose topical corticosteroid inhibits IL-5 and IL-13 in nasal lavage following grass pollen challenge. AB - BACKGROUND: Nasal lavage is a noninvasive method of obtaining inflammatory exudates following nasal allergen challenge (NAC), and permits cells and released mediators to be evaluated. OBJECTIVE: To determine the effects of a single dose of topical steroid on eosinophils and levels of chemokines and cytokines in nasal lavage fluid following NAC in patients with allergic rhinitis. METHODS: Patients with grass pollen seasonal allergic rhinitis (n = 32) out of the allergy season received either nasal budesonide (100 microg per nostril) or matched placebo before allergen challenge in a double blind two-way crossover design. A semi automated mixed bead array system was employed to measure multiple chemokines and cytokines in small volumes (50 microl) of nasal lavage supernatants. RESULTS: Following NAC there was a rapid onset of nasal symptoms together with nasal eosinophilia, and the appearance of IL-5 and IL-13 in lavages between 4 and 8 h. Elevated levels of eotaxin, RANTES, IL-8 and MCP-1 were also detected following allergen challenge. A single dose of nasal budesonide caused a decrease in symptoms (P < 0.05) and nasal eosinophils (P < 0.05) with selective abrogation of IL-5 and IL-13 responses (P < 0.05), but a lack of effect on levels of eotaxin, RANTES, IL-8 and MCP-1. CONCLUSION: This study suggests that a single dose of nasal steroid has the capacity to selectively abolish IL-5 and IL-13 responses following NAC. This model should be convenient for testing novel anti inflammatory and immunoregulatory agents intended for the treatment of allergic rhinitis. PMID- 16266386 TI - New cohorts of naive T cells exacerbate ongoing allergy but can be suppressed by regulatory T cells. AB - Although as pretreatment oral tolerance is a potent means to achieve systemic suppression, its application in ongoing disease is controversial. Here we propose that availability of naive T cells may critically determine whether immunological tolerance is achieved during ongoing antigenic reactivity. Infusion of naive antigen-specific T cells into mice directly prior to eliciting a secondary Th2 response induces these naive cells to actively engage in the antigenic response despite presence of established memory. Naive antigen-specific T-cells divided faster, produced more interleukin (IL)-2, IL-4 and IL-5 and enhanced immunoglobulin E (IgE) release during a secondary Th2 response, compared with naive T cells that were infused prior to a primary response. Despite such contribution by new cohorts of naive T cells co-infusion of mucosal Tr together with naive T cells could suppress enhanced IgE release during a secondary Th2 response. We conclude that naive T cells contribute to a secondary Th2 response and although they can still be suppressed in the presence of sufficient numbers of mucosal Tr, they may interfere with potential therapeutic application of mucosal tolerance. PMID- 16266387 TI - Fast foods - are they a risk factor for asthma? AB - BACKGROUND: Lifestyle changes over the last 30 years are the most likely explanation for the increase in allergic disease over this period. AIM: This study tests the hypothesis that the consumption of fast food is related to the prevalence of asthma and allergy. METHODS: As part of the International Study of Asthma and Allergies in Childhood (ISAAC) a cross-sectional prevalence study of 1321 children (mean age = 11.4 years, range: 10.1-12.5) was conducted in Hastings, New Zealand. Using standard questions we collected data on the prevalence of asthma and asthma symptoms, as well as food frequency data. Skin prick tests were performed to common environmental allergens and exercise-induced bronchial hyperresponsiveness (BHR) was assessed according to a standard protocol. Body mass index (BMI) was calculated as weight/height2 (kg/m2) and classified into overweight and obese according to a standard international definition. RESULTS: After adjusting for lifestyle factors, including other diet and BMI variables, compared with children who never ate hamburgers, we found an independent risk of hamburger consumption on having a history of wheeze [consumption less than once a week (OR = 1.44, 95% CI: 1.06-1.96) and 1+ times a week (OR = 1.65, 95% CI: 1.07-2.52)] and on current wheeze [consumption less than once a week (OR = 1.17, 95% CI: 0.80-1.70) and 1+ times a week (OR = 1.81, 95% CI: 1.10-2.98)]. Takeaway consumption 1+ times a week was marginally significantly related to BHR (OR = 2.41, 95% CI: 0.99-5.91). There was no effect on atopy. CONCLUSIONS: Frequent consumption of hamburgers showed a dose-dependent association with asthma symptoms, and frequent takeaway consumption showed a similar association with BHR. PMID- 16266389 TI - Anaphylaxis by fruits of the Fagaceae family: acorn and chestnut. PMID- 16266388 TI - Occupational eosinophilic bronchitis. PMID- 16266390 TI - Occupational asthma due to bethabara wood dust. PMID- 16266391 TI - Selective immediate hypersensitivity to cefodizime. PMID- 16266392 TI - Is semen quality affected by male body fat distribution? AB - The aim of this study was to examine the relationship of semen parameters, sexual function-related hormones and waist/hip ratio. Eighty-one selected patients presenting with infertility were examined. Weight, height, waist circumference and hip circumference were measured, and reproduction-related hormone levels were determined. Semen was analysed by conventional methods. Semen volume, sperm concentration, motility, total sperm count, total motile sperm cell number, rapid progressive motile sperm count and reproduction-related hormone levels [follicle stimulating hormone, luteinizing hormone, prolactin, testosterone, 17beta oestradiol and sexual hormone-binding globulin (SHBG)]. Significant correlations were found: (i) weight, waist circumference and hip circumference versus testosterone level, SHBG level, and testosterone/17beta-oestradiol ratio; (ii) hip circumference versus sperm concentration; (iii) waist circumference and hip circumference versus sperm count, total motile sperm cell number and rapid progressive motile sperm count; (iv) weight versus total sperm count and total motile sperm cell number; (v) waist circumference and hip circumference versus prolactin level (positively) and SHBG (negatively); (vi) waist circumference and waist/hip ratio versus semen volume. It can be concluded that the waist/hip ratio is correlated with the reproductive hormone levels. Although both the waist circumference and hip circumference correlated with the semen characteristics, the waist/hip ratio did not. PMID- 16266393 TI - Bacterial flora of the low male genital tract in patients consulting for infertility. AB - The physiological aerobic bacterial flora of the low male genital tract was determined. This prospective study was performed on 600 semen specimens collected from 543 asymptomatic males consulting for infertility. Semen cultures were sterile in 28.8%, with a polymicrobial flora and/or absence or low titres of Ureaplasma urealyticum in 49.3%, and with one or two aerobic and facultative bacteria > or =1 x 10(3) CFU ml(-1) and/or U. urealyticum with titres > or =10(4) CCU ml(-1) (colour changing units) in 21.8%. In standard aerobic cultures, Gardnerella vaginalis was the most commonly isolated species (26.1%), followed by coagulase-negative staphylococci (15.7%) and Streptococcus anginosus (14.2%). Ureaplasma urealyticum was absent in 84.5% of semen samples, but when recovered, high (> or =10(4) CCU ml(-1)) and low titres (< or =10(3) CCU ml(-1)) were counted in 7.2% and 8.3% respectively. Of 48 patients, the follow-up of semen cultures showed marked variations in time. This study shows that (i) there was no relationship between the bacterial flora and the leucocytospermia; (ii) low titres of U. urealyticum in semen were not associated with a disturbance of the ecosystem; (iii) the critical threshold for U. urealyticum should be raised to > or =10(4) CFU ml(-1) and (iv) a positive semen culture should be repeated before any treatment. PMID- 16266394 TI - Nitric oxide induces acrosome reaction in cryopreserved bovine spermatozoa. AB - The aim of this work was to study the effect of nitric oxide on acrosome reaction (AR) and the participation of protein kinases and reactive oxygen species in the AR of cryopreserved bovine spermatozoa. Spermatozoa were capacitated in Tyrode's albumin lactate pyruvate medium with heparin (10 IU ml(-1)) and then incubated with different concentrations of sodium nitroprusside (SNP) (1-200 micromol l( 1)). Methylene blue and haemoglobin were used to confirm the role of nitric oxide as an inducer of the AR. The participation of protein kinase A (PKA) , protein kinase C (PKC) and protein tyrosine kinase was evaluated using specific inhibitors of these enzymes (H-89, 50 micromol l(-1); bisindolylmaleimide I, 0.1 micromol l(-1) and genistein, 3 micromol l(-1)). The role of hydrogen peroxide or superoxide anion was evaluated by incubation with catalase or superoxide dismutase respectively. AR percentages were determined by the fluorescence technique with chlortetracycline. The highest levels of AR were obtained in capacitated spermatozoa treated with 5-200 micromol l(-1) SNP (24.8 +/- 1.8%). The presence of PKA, PKC and protein tyrosine kinase inhibitors likewise decreased AR percentages. The addition of superoxide dismutase had no effect on the AR level but catalase completely blocked it. These results indicate that nitric oxide induces AR in capacitated spermatozoa involving hydrogen peroxide and the participation of PKA, PKC and protein tyrosine kinase as part of the signal transduction mechanism which lead to the AR in cryopreserved bovine spermatozoa. PMID- 16266395 TI - Serum inhibin B and follicle-stimulating hormone levels as markers in the evaluation of azoospermic men: a comparison. AB - Inhibin B is a glycoprotein hormone produced mainly by Sertoli cells of the testes in the adult male. It selectively suppresses the secretion of pituitary follicle-stimulating hormone (FSH) and has local paracrine actions in the testes. Its measurement is useful for investigating the role of inhibin B in male gonadal dysfunction. The objective of this study was to investigate the efficacy of serum inhibin B in men with nonobstructive azoospermia in comparison with FSH. Serum concentration of FSH was measured using microparticle enzyme immunoassay, inhibin B by specific solid phase sandwich enzyme-linked immunosorbent assay in men with nonobstructive azoospermia (n = 46) and control fertile men (n = 5). Mean inhibin B and FSH level was 104.6 pg ml(-1) and 4.0 mIU ml(-1) in control men whereas the value for nonobstructive azoospermic men was 17.06 pg ml(-1) and 31.1 mIU ml(-1) respectively. Inhibin B and FSH levels were significantly different in azoospermia than controls (P < 0.0001). There were six cases of nonobstructive azoospermia with normal inhibin B. Testicular histology did not find any evidence of spermatogenesis in three cases with normal inhibin B. This demonstrated that inhibin B was not a superior predictor for testicular function in our study. PMID- 16266396 TI - Tunica albuginea overlapping: a novel technique for the treatment of erectile dysfunction. AB - Tunica albuginea (TA) in venogenic erectile dysfunction (VED) was found subluxated and flabby because of degeneration and atrophy of its collagen fibres. This had apparently led to derangement of TA veno-occlusive mechanism. We investigated the hypothesis that overlapping of the subluxated and flabby TA would achieve a competent veno-occlusive mechanism during erection. Tunical overlapping was performed in nine VED patients (age 35.6 +/- 1.6 years). Intracorporal pressure (ICP) was measured pre- and postoperatively. After penile degloving, TA on lateral penile aspect was divided along whole length of corpus cavernosum (CC) and tunical double-breasting for 1-1 1/2 cm was performed. A biopsy was taken from TA and stained with haematoxylin and eosin and Masson's trichrome. Clinical efficiency of the operation was evaluated after 6 months. ICP increased (P < 0.01) postoperatively in the nine patients. The increase was maintained during follow-up period in eight patients and decreased to preoperative level in one. Six months after operation, the eight patients had significantly (P < 0.01) improved scores for the erectile function domain over the preoperative scores. Microscopic examination of TA biopsies showed atrophy of the collagen fibres. Tunical overlapping aims at correction of TA flabbiness, corporal tissue support and improving of veno-occlusive mechanism. PMID- 16266397 TI - Are antisperm antibodies indeed sperm-specific? AB - Antisperm antibodies (ASA) are present in a high percentage of infertile patients. The development of ASA in the male depends on the sequestration of antigens on germ cells, the testis being an immune privileged region. In this study, we investigated the specificity of ASA binding to sperm antigens by exposing a number of organ preparations to ASA. In none of the organ homogenates was a significant difference between the results of the Western blot with ASA containing fluids, neither serum nor seminal plasma, and those without ASA observed. We conclude from our results that the ASA tested in our study obviously are sperm-specific. ASA as an autoimmune are thus restricted to spermatozoa. The antigens are suitable for trials in immune therapy, they may also serve for the development of antigen-specific diagnosis and treatment in infertility. They also indicate cognate antigens of possible immune contraceptive agents. PMID- 16266398 TI - Role of granulocyte elastase and interleukin-6 in the diagnosis of male genital tract inflammation. AB - Chronic genital tract inflammations are a frequent cause or at least a concomitant factor of male fertility disturbances. The diagnosis is difficult because of the mostly asymptomatic course of the disease. Therefore, determination of biochemical markers of inflammation in addition to the number of leucocytes in the seminal plasma has been recommended. The aim of the study was to find out whether determination of granulocyte elastase and interleukin-6 provide comparable and reliable results with regard to diagnosis of genital tract inflammation; in addition, the association between genital tract inflammation and semen quality should be evaluated with special focus on potentially disturbed sperm functions like sperm motility and DNA integrity. In a prospective study, the concentrations of interleukin-6 (IL-6) and granulocyte elastase were determined in seminal plasma samples from 340 patients to investigate the relationship with other parameters of genital tract inflammation such as the number of peroxidase-positive cells and conventional semen parameters. Microbiological investigations were included. As post-testicular inflammatory influences may cause sperm DNA damage, the correlation between IL-6 and elastase and DNA integrity was evaluated by the sperm chromatin structure assay. IL-6 and elastase were significantly correlated both with each other (P < 0.01) and the number of peroxidase-positive cells (P < 0.01). IL-6 showed a highly significant negative correlation with sperm vitality (P < 0.01) and a significant negative correlation with sperm motility (P < 0.05). Elastase concentrations were highly significantly associated with the number of peroxidase-positive cells (P < 0.01) and negatively correlated with sperm vitality (P < 0.01). Moreover, there were significantly negative correlations with sperm motility (P < 0.05), progressive motility according to WHO a quality (P < 0.05) as well as sperm morphology (P < 0.05). In addition, a significant negative correlation was observed between elastase concentrations and percentage of spermatozoa with intact DNA, which may suggest the use of anti-inflammatory treatment. It can be concluded that both IL 6 and granulocyte elastase are useful and suitable as markers for silent genital tract inflammation; in contrast to previous contributions there were clear correlations of IL-6 and granulocyte elastase with sperm parameters, the relationship of elastase with semen quality being more marked. Moreover, the results of the study confirm the need for a change of the threshold value of peroxidase-positive cells according to WHO definition to lower levels for definition of silent genital tract inflammation. PMID- 16266401 TI - The significant increase in cardiovascular disease risk in APOEepsilon4 carriers is evident only in men who smoke: potential relationship between reduced antioxidant status and ApoE4. AB - Data from 1668 men (316 cardiovascular disease events) from the Framingham Offspring Study was reanalysed, specifically examining APOE:smoking interactions. Overall hazard ratio (HR) for smoking was 1.95 (1.52, 2.50) compared to non smokers. Using epsilon3/3 as a referent group, in non-smokers HRs for epsilon2 carriers (epsilon2+; 1.04 (0.61, 1.76) and epsilon4 carriers (epsilon4+; 1.04 (0.70, 1.54) showed no major risk increase. In smokers, HRs were 1.96 (1.26, 2.78) in epsilon3epsilon3 men, 3.46 (2.14, 5.60; p = 0.09 for interaction) in epsilon2+ and 3.81 (2.49, 5.84; p = 0.01 for interaction), with a significant interaction between daily cigarette consumption and APOE genotype on risk (p = 0.03). The potential mechanism for this APOEepsilon4:smoking interaction was examined in a second study of 728 Caucasian patients with diabetes, where markers of reactive oxygen species were available. APOE genotype was not associated with plasma OX-LDL or total antioxidant status (TAOS) in non-smokers. However, in smokers epsilon4+ had 26.7% higher plasma OX-LDL than other genotypes (APOE:smoking interaction p = 0.04), while epsilon2+ had 28.4% higher plasma TAOS than epsilon3epsilon3 and epsilon4+ combined (APOE:smoking interaction p = 0.026). Although direct extrapolation needs to be considered with caution, these results identify that the cardiovascular disease risk-raising effect of epsilon4+ is confined to smokers, and a feasible mechanism is presented by the reduced antioxidant capacity/increased OX-LDL of apoE4. PMID- 16266403 TI - The association of mitochondrial DNA 5178 C > a polymorphism with plasma lipid levels among three ethnic groups. AB - Mitochondria are eukaryotic cytoplasmic organelles responsible for oxidative phosphorylation. The C to A nucleotide transversion in the NADH dehydrogenase subunit 2 (MT-ND2) coding region of mitochondrial DNA has been reported to be associated with plasma lipid levels, adult onset diseases and longevity. We have examined the role of this polymorphism in relation to plasma lipid levels and age in a total of 713 healthy individuals belonging to 3 ethnic groups in Singapore. The frequency of the A allele was significantly higher (p < 0.05) among the Chinese (0.15) in comparison to the Malays (0.05) and Indians (0.02). No significant difference in the frequency of the allele was observed between healthy and coronary artery disease subjects, and between age-stratified subjects. We found that the polymorphism is significantly associated in an ethnic and gender-specific manner with plasma apoB levels in the Chinese males (p < 0.05). This is the first epidemiological report of the mt5178 C > A polymorphism and its association with plasma lipid levels in Asian populations outside Japan. PMID- 16266402 TI - Association of common CRP gene variants with CRP levels and cardiovascular events. AB - C-reactive protein (CRP) is a well-documented marker of atherosclerotic cardiovascular disease risk. We resequenced CRP to identify a comprehensive set of common SNP variants, then studied and replicated their association with baseline CRP level among apparently healthy subjects in the Women's Health Study (WHS; n = 717), Pravastatin Inflammation/CRP Evaluation trial (PRINCE; n = 1,110) and Physicians' Health Study (PHS; n = 509) cohorts. The minor alleles of four SNPs were consistently associated in all three cohorts with higher CRP, while the minor alleles of two SNPs were associated with lower CRP (p < 0.05 for each). Single marker and haplotype analysis in all three cohorts were consistent with functional roles for the 5'-flanking triallelic SNP -286C>T>A and the 3'-UTR SNP 1846G>A. None of the SNPs associated with higher CRP were associated with risk of incident myocardial infarction (MI) or ischemic stroke in a prospective, nested case-control study design from the PHS cohort (610 case-control pairs). One SNP, 717A>G, was unrelated to CRP levels but associated with decreased risk of MI (p = 0.001). Taken together, these data imply significant interactions between both genetic and environmental contributions to the increased CRP levels that predict a greater risk of future atherothrombotic events in epidemiological studies. PMID- 16266404 TI - Possible association of the human KCNE1 (minK) gene and QT interval in healthy subjects: evidence from association and linkage analyses in Israeli families. AB - QT interval prolongation is associated with increased risk of sudden and non sudden cardiac death. Potassium channel gene variants are associated with inherited long QT syndromes. Using linkage and association analyses, we investigated whether variants in the potassium channel subunit KCNE1 are associated with QTc intervals in an unselected population sample of 80 kindreds living in kibbutz settlements in Israel. Variance-component linkage analysis revealed weak evidence of linkage of KCNE1 polymorphisms with QTc intervals. Family-based association analysis showed a significant association between the G38S polymorphism and QTc interval. Further quantitative trait association analysis demonstrated a significant residual heritability component (h(2)= 0.33), and that the effect of the G38S variant allele is modified by gender. Estimated maximum likelihood parameters from these models indicated that male gender, age, hypertension, diabetes, hypercholesterolemia, fibrinogen and BMI were positively associated with QTc interval; level of education and cigarette smoking showed an inverse association. Both erythrocyte membrane n-6 and n-3 fatty acids showed a significant inverse association with QTc interval. While more than 15.8% of QTc variability was contributed by covariates, another 4.7% was explained by dietary factors, the G38S polymorphism explained 2.2%, and approximately 36% was explained by polygenes. An in silico analysis showed also that the novel V80 SNP, another KCNE1 synonymous variant, abolishes the recognition for a splicing enhancer, which may lead to an increased effect of the G38S mutation. These results demonstrate that, in addition to polygenic background, dietary factors and other covariables, the KCNE1 G38S variant is involved in determining QTc levels in this population-based sample of families. PMID- 16266405 TI - ATM gene founder haplotypes and associated mutations in Polish families with ataxia-telangiectasia. AB - Ataxia-telangiectasia (A-T) is an early onset autosomal recessive ataxia associated with characteristic chromosomal aberrations, cell cycle checkpoint defects, cancer susceptibility, and sensitivity to ionizing radiation. We utilized the protein truncation test (PTT), and single strand conformation polymorphism (SSCP) on cDNA, as well as denaturing high performance liquid chromatography (dHPLC) on genomic DNA (gDNA) to screen for mutations in 24 Polish A-T families. Twenty-six distinct Short Tandem Repeat (STR) haplotypes were identified. Three founder mutations accounted for 58% of the alleles. Three quarters of the families had at least one recurring (shared) mutation, which was somewhat surprising given the low frequency of consanguinity in Poland. STR haplotyping greatly improved the efficiency of mutation detection. We identified 44 of the expected 48 mutations (92%): sixty-nine percent were nonsense mutations, 23% caused aberrant splicing, and 5% were missense mutations. Four mutations have not been previously described. Two of the Polish mutations have been observed previously in Amish and Mennonite A-T patients; this is compatible with historical records. Shared mutations shared the same Single Nucleotide Polymorphism (SNP) and STR haplotypes, indicating common ancestries. The Mennonite mutation, 5932 G>T, is common in Russian A-T families, and the STR haplovariants are the same in both Poland and Russia. Attempts to correlate phenotypes with genotypes were inconclusive due to the limited numbers of patients with identical mutations. PMID- 16266406 TI - Temporal mitochondrial DNA variation in the Basque Country: influence of post neolithic events. AB - The Basque population has been considered an outlier in a large number of genetic studies, due to its hypothesized antiquity and greater genetic isolation. The present paper deals with an analysis of the mtDNA variability of the historical population of Aldaieta (VI-VII c. AD; Basque Country) which, together with genetic data existing for other prehistoric populations of the Basque Country (4,500-5,000 YBP), permits an appraisal of the hypotheses proposed for the origin of the genetic differentiation of the Basque population. Given that this is an aDNA study, application has been made both of standard precautions, to avoid contamination, and of authentication criteria (analysis of duplicates, replication in an independent laboratory, quantification of target DNA, sequencing and cloning of PCR products). The variability of the mtDNA haplogroups of the historical population of Aldaieta falls within the range of the present day populations of Europe's Atlantic fringe, whereas the prehistoric populations of the Basque Country display clear differentiation in relation to all others. Consequently, we suggest that between 5,000-1,500 YBP approximately, there may have been gene flow amongst the western European populations that homogenised mtDNA lineages. PMID- 16266407 TI - Diversity and divergence among the tribal populations of India. AB - Tribal populations of the Indian subcontinent have been of longstanding interest to anthropologists and human geneticists. To investigate the relationship of Indian tribes to Indian castes and continental populations, we analyzed 45 unlinked autosomal STR loci in 9 tribal groups, 8 castes, and 18 populations from Africa, Europe and East Asia. South Indian tribal populations demonstrate low within-population heterozygosity (range: 0.54 - 0.69), while tribal populations sampled further north and east have higher heterozygosity (range: 0.69 - 0.74). Genetic distance estimates show that tribal Indians are more closely related to caste Indians than to other major groups. Between-tribe differentiation is high and exceeds that for eight sub-Saharan African populations (4.8% vs. 3.7%). Telugu-speaking populations are less differentiated than non-Telugu speakers (F(ST): 0.029 vs. 0.079), but geographic distance was not predictive of genetic affinity between tribes. South Indian tribes show significant population structure, and individuals can be clustered statistically into groups that correspond with their tribal affiliation. These results are consistent with high levels of genetic drift and isolation in Indian tribal populations, particularly those of South India, and they imply that these populations may be potential candidates for linkage disequilibrium and association mapping. PMID- 16266408 TI - Assessment of the role of genetic polymorphism in venous thrombosis through artificial neural networks. AB - PURPOSE: To assess the role of genetic polymorphisms in venous thrombosis events (VTE) using Artificial Neural Networks (ANNs), a model for solving non-linear problems frequently associated with complex biological systems, due to interactions between biological, genetic and environmental factors. METHODS: A database was generated from a case-control study of venous thrombosis, using 238 patients and 211 controls. The database of 64 variables included age, gender and a panel of 62 genetic variants. Three different ANNs were compared, with logistic regression for the accuracy of predicting cases and controls. RESULTS: ANNs yielded a better performance than the logistic regression algorithm. Indeed, through ANNs models, the 62 variables related to genetic variants were first reduced to a set of 9, and then of 3 (MTHFR 677 C/T, FV arg506gln, ICAM1 gly214arg). CONCLUSIONS: The findings of this study illustrate the power of ANN in evaluating multifactorial data, and show that the different sensitivities of the models of elaboration are related to the characteristics of the data. This may contribute to a better understanding of the role played by genetic polymorphisms in VTE, and help to define, if possible, a test panel of genetic variants to estimate an individual's probability of developing the disease. PMID- 16266409 TI - Interaction between two quantitative trait loci affects fetal haemoglobin expression. AB - The biological mechanisms controlling complex quantitative traits are likely to be affected by interactions between genetic factors, sometimes referred to as epistasis. The identification of interacting loci through genetic analyse faces many challenges, and few examples of replicated findings of interaction exist for humans and model system organisms. The replication of an interaction, or the non independence, of two quantitative trait loci (QTL) affecting the developmental switch from the expression of fetal to adult haemoglobin is reported here. Fetal haemoglobin expression in adults is a highly heritable, yet complex, phenotype. Using a sample of 874 dizygotic twin pairs of European descent, we found linkage to a QTL on chromosome 8 to be conditional on the twin pairs' genotypes at a polymorphism in the beta-globin complex; an interaction originally identified in a large Asian Indian kindred. The beta-globin polymorphism has been previously shown to be associated with fetal haemoglobin levels in adults. This study reports the first known replication of a genetic interaction between QTLs influencing a complex human trait. PMID- 16266410 TI - Tests of association between quantitative traits and haplotypes in a reduced dimensional space. AB - Candidate gene association tests are currently performed using several intragenic SNPs simultaneously, by testing SNP haplotype or genotype effects in multifactorial diseases or traits. The number of haplotypes drastically increases with an increase in the number of typed SNPs. As a result, large numbers of haplotypes will introduce large degrees of freedom in haplotype-based tests, and thus limit the power of the tests. In this study we propose using the principal component method to reduce the dimension, and then construct association tests on the lower-dimensional space to test the association between haplotypes and a quantitative trait using population-based samples. The proposed method allows ambiguous haplotypes. We use simulation studies to evaluate the type I error rate of the tests, and compare the power of the proposed tests with that of the tests without dimension reduction, and the tests with dimension reduction by merging rare haplotypes. The simulation results show that the proposed tests have correct type I error rates and are more powerful than other tests in most cases considered in our simulation studies. PMID- 16266411 TI - SNP selection for association studies: maximizing power across SNP choice and study size. AB - Selection of single nucleotide polymorphisms (SNPs) is a problem of primary importance in association studies and several approaches have been proposed. However, none provides a satisfying answer to the problem of how many SNPs should be selected, and how this should depend on the pattern of linkage disequilibrium (LD) in the region under consideration. Moreover, SNP selection is usually considered as independent from deciding the sample size of the study. However, when resources are limited there is a tradeoff between the study size and the number of SNPs to genotype. We show that tuning the SNP density to the LD pattern can be achieved by looking for the best solution to this tradeoff. Our approach consists of formulating SNP selection as an optimization problem: the objective is to maximize the power of the final association study, whilst keeping the total costs below a given budget. We also propose two alternative algorithms for the solution of this optimization problem: a genetic algorithm and a hill climbing search. These standard techniques efficiently find good solutions, even when the number of possible SNPs to choose from is large. We compare the performance of these two algorithms on different chromosomal regions and show that, as expected, the selected SNPs reflect the LD pattern: the optimal SNP density varies dramatically between chromosomal regions. PMID- 16266412 TI - Multiple testing in the context of haplotype analysis revisited: application to case-control data. AB - We have lately presented a testing procedure for family data which accounts for the multiple testing problem that is induced by the enormous number of different marker combinations that can be analyzed in a set of tightly linked markers. Most methods of haplotype based association analysis already require simulations to obtain an uncorrected P value for a specific marker combination. As shown before, it is nevertheless not necessary to carry out nested simulations to obtain a global P value that properly corrects for the multiple testing of different marker combinations without neglecting the dependency of the tests. We have now implemented this approach for case-control data in our program FAMHAP, as this data structure currently plays a dominant role in the field. We consider different ways to deal with phase ambiguities and two different statistical tests for the underlying single marker combinations to obtain uncorrected P values. One test statistic is chi-square based, the other is a haplotype trend regression. The performance of these different tests in the multiple testing situation is investigated in a large simulation study. We obtain a considerable gain in power with our global P values as opposed to Bonferroni corrected P values for all suggested test statistics. Good power was obtained both with the haplotype trend regression approach as well as with the simpler chi-square based test. Furthermore, we conclude that the better strategy to deal with phase ambiguities is to assign to each individual its list of weighted haplotype explanations, rather than to assign to each individual its most likely haplotype explanation. Finally, we demonstrate the usefulness of our approach by a real data example. PMID- 16266415 TI - Guest editorial. PMID- 16266413 TI - The peopling of modern Bosnia-Herzegovina: Y-chromosome haplogroups in the three main ethnic groups. AB - The variation at 28 Y-chromosome biallelic markers was analysed in 256 males (90 Croats, 81 Serbs and 85 Bosniacs) from Bosnia-Herzegovina. An important shared feature between the three ethnic groups is the high frequency of the "Palaeolithic" European-specific haplogroup (Hg) I, a likely signature of a Balkan population re-expansion after the Last Glacial Maximum. This haplogroup is almost completely represented by the sub-haplogroup I-P37 whose frequency is, however, higher in the Croats (approximately 71%) than in Bosniacs (approximately 44%) and Serbs (approximately 31%). Other rather frequent haplogroups are E (approximately 15%) and J (approximately 7%), which are considered to have arrived from the Middle East in Neolithic and post-Neolithic times, and R-M17 (approximately 14%), which probably marked several arrivals, at different times, from eastern Eurasia. Hg E, almost exclusively represented by its subclade E-M78, is more common in the Serbs (approximately 20%) than in Bosniacs (approximately 13%) and Croats (approximately 9%), and Hg J, observed in only one Croat, encompasses approximately 9% of the Serbs and approximately 12% of the Bosniacs, where it shows its highest diversification. By contrast, Hg R-M17 displays similar frequencies in all three groups. On the whole, the three main groups of Bosnia-Herzegovina, in spite of some quantitative differences, share a large fraction of the same ancient gene pool distinctive for the Balkan area. PMID- 16266416 TI - A consultation leaflet to improve an older patient's involvement in general practice care: a randomized trial. AB - OBJECTIVE: To evaluate the effects of a programme to enhance the involvement of older patients in their consultations in general practice. DESIGN: Cluster randomized trial, in which data was collected from different cohorts. SETTING AND PARTICIPANTS: Twenty-five general practices in the south-east part of the Netherlands and their patients aged 70 years and over. INTERVENTION: Patients in the intervention group received a leaflet to help them prepare for the consultation. General practitioners (GPs) received an outreach visit to optimize older patients' involvement when visiting their GP. Patients in the control group received usual care. MAIN OUTCOME MEASURES: Questionnaires measuring involvement (COMRADE), enablement (Patient Enablement Index) and satisfaction with their care (EUROPEP). RESULTS: Pre-intervention 315 patients and post-intervention 263 patients were included. Subjects were satisfied with their involvement and the GP's behaviour during the consultation. No differences in effect as a result of the leaflet on involvement, enablement or satisfaction were found between the intervention and the control group. Of 318 patients who received the leaflet and visited their GP in the intervention period, 47 patients used the leaflet. These users were more accustomed to prepare themselves for consultations. Users reported more psychological problems than non-users. CONCLUSIONS: No relevant effects of the implementation programme on involvement, enablement or satisfaction were found. Other strategies are needed to enhance involvement of older patients in their care. Alternatively, older patients may perceive themselves sufficiently involved. PMID- 16266417 TI - Informed consent for mammography screening: modelling the risks and benefits for American women. AB - INTRODUCTION: In order to facilitate informed decision making, women require information on the probabilities of different outcomes with mammography screening. This paper derives these probabilities for a US population and illustrates them visually in a readily understandable format. METHODS: Probabilities of the breast cancer mortality, all cause mortality and further investigation are derived from published data on mortality from breast cancer and published estimates of effectiveness using a life-table method. Probabilities are calculated of surviving to age 75 from age 40 with and without two-yearly mammography screening from age 40 and age 50. Probabilities are also calculated that a woman will be referred for further assessment or biopsy or die from breast cancer despite screening. To avoid being misled, these outcomes are presented in the form of a single decision aid illustrating the outcomes for 1000 women choosing each alternative: mammography screening or no mammography screening. RESULTS: Of 1000 women undergoing two-yearly mammography screening from age 40 an additional four (3.7 per 1000) will reach the age of 75; of the survivors 514 will be referred for further investigation and 138 will undergo biopsy. Of 1000 women screened from age 50 an additional three (3.3 per 1000) will reach age 75; of the survivors 408 will be referred for further investigation and 94 will undergo biopsy. Mammography from age 40 to 49 reduces mortality by 0.4 in 1000. This information is readily presented visually. CONCLUSIONS: It is possible to provide realistic estimates of the effects of mammography screening on mortality in a readily understandable format. Women require this information if they are to make informed choices about mammography screening. PMID- 16266418 TI - The rhetoric of informed choice: perspectives from midwives on intrapartum fetal heart rate monitoring. AB - OBJECTIVE: To investigate midwives' attitudes, values and beliefs on the use of intrapartum fetal monitoring. DESIGN: Qualitative, semi-structured interviews. SUBJECTS AND SETTING: Fifty-eight registered midwives in two hospitals in the North of England. RESULTS: In this paper two main themes are discussed, these are: informed choice, and the power of the midwife. Midwives favoured the application of informed choice and shared a unanimous consensus on the definition. However, the idealistic perception of informed choice, which included contemporary notions of empowerment and autonomy for women expressing an informed choice, was not reportedly translated into practice. Midwives had to implement informed choice on intrapartum fetal monitoring within a competing set of health service agendas, i.e. medically driven protocols and a political climate of actively managed childbearing. This resulted in the manipulation of information during the midwives' interactions with women. This ultimately meant that the women often got the choice the midwives wanted them to have. CONCLUSIONS: The information that a midwife imparts may consciously or subconsciously affect the woman's uptake and understanding of information. Therefore, the midwife has a powerful role to play in balancing the benefits and risk ratios applicable to fetal heart rate monitoring. However, a deeply ingrained pre-occupation with technological methods of intrapartum fetal monitoring over many years has made it difficult for midwives to offer alternative forms of monitoring. This has placed limits on the facilitation of informed choice and autonomous decision making for women. PMID- 16266419 TI - Exploring the role of self-management programmes in caring for people from culturally and linguistically diverse backgrounds in Melbourne, Australia. AB - BACKGROUND: Chronic disease self-management programmes are now an important adjunct to the treatment and care of Australians with chronic illnesses. Most programmes are delivered in English and cater for 'Anglo' views of health and illness. The Peer-Led Self-Management of Chronic Illness Project was funded by the National Health and Medical Research Council (NHMRC) to test the hypothesis that the Stanford University Chronic Disease Self-Management Program would improve health outcomes for people from the Vietnamese, Greek, Chinese and Italian communities in Melbourne's north-eastern suburbs. OBJECTIVE: To examine the extent to which the programme required modification so that the concepts associated with self-management programmes have relevance to the health behaviours of people with chronic illness from the above communities. METHODS: Four focus groups facilitated in English, using interpreters. RESULTS: There was wide understanding of the concepts employed in self-management programmes. Literacy problems emerged as the major obstacle to participating in unmodified programmes. CONCLUSION: The conceptual aspects of the programme require less modification than originally predicted, but the programme requires sensitive modification so that it is accessible to people with low literacy levels. PMID- 16266420 TI - Meeting the educational needs of people at risk of diabetes-related amputation: a vignette study with patients and professionals. AB - OBJECTIVE: To investigate how patients and professionals view the role of advice in diabetes foot care, in order to inform educational practice. DESIGN: A qualitative study based upon one-to-one interviews using a vignette technique with patients and professionals. SETTING AND PARTICIPANTS: Interviews with 15 patients with diabetes-related foot complications attending diabetes foot clinics, and 15 health professionals across two hospital sites and two primary care trusts. RESULTS: There were similarities between issues raised by patients and professionals, with differences in emphasis. An important factor was patient understanding and acceptance of the implications of diabetes from diagnosis. Many patients reported a lack of early understanding that may reflect inadequate provision or acceptance of advice. Development of positive relationships with health professionals was important for patients and encouraged shared understanding. Use of the vignette technique highlighted the gap between patient knowledge and action, with patients and professionals emphasizing their own responsibilities in relation to reduction of risk through education and action. Responses suggest that early positive interactions with appropriately trained professionals are needed to help patients assimilate advice into everyday routines. CONCLUSIONS: For effective preventive care, patients need to understand how diabetes impacts on their health. Foot care education should begin early, be patient-centred and delivered with empathy by professionals whom the patients trust. The findings reflect patient and professional expectations in educational practice, and therefore have relevance for other chronic conditions for which much education and advice is related to self-care. PMID- 16266421 TI - Patient priorities in colorectal cancer screening decisions. AB - BACKGROUND: Colorectal cancer screening guidelines in the United States recommend that decisions about screening should incorporate patient preferences, but little is known about how patients make the trade-offs inherent in choosing one of the five currently recommended screening programmes. STUDY POPULATION: Forty-eight primary care patients at average risk for colorectal cancer who completed an experimental shared decision-making intervention based on a multicriteria decision analysis. METHODS: Descriptive analysis of priorities assigned to decision criteria describing the advantages and disadvantages of the five currently recommended colorectal screening programmes in the United States. Criteria were divided into four major criteria - avoid cancer, avoid screening side-effects, avoid false positive test results and the combined importance of other considerations - and three subcriteria: the number of screening tests, test preparation and the test itself. Cluster analysis was used to identify common combinations of priorities within each set of criteria. RESULTS: Patients assigned widely variable priorities to both the criteria and subcriteria: the average range of priorities was 46 on a 100 point priority scale. Cluster analysis identified six different combinations of priorities for the major criteria and four for the subcriteria. The differences in priorities assigned to both the criteria and subcriteria in the clusters were statistically significant with P < 0.0001. CONCLUSIONS: Even within a small group of patients, preferences vary widely regarding trade-offs involved in choosing among the currently recommended colorectal cancer screening programmes in the United States. These results provide empiric support for recommendations to utilize a shared decision making process when making colorectal cancer screening decisions and highlight the need for additional research into how average risk patients view the trade offs inherent in choosing a colorectal cancer screening programme. PMID- 16266422 TI - Patient, consumer, client, or customer: what do people want to be called? AB - OBJECTIVE: To clarify preferred labels for people receiving health care. BACKGROUND: The proper label to describe people receiving care has evoked considerable debate among providers and bio-ethicists, but there is little evidence as to the preferences of the people involved. DESIGN: We analysed dictionary definitions as to the derivation and connotations of such potential labels as: patient, client, customer, consumer, partner and survivor. We then surveyed outpatients from four clinical populations in Ontario, Canada about their feelings about these labels. SETTING AND PARTICIPANTS: People from breast cancer (n = 202), prostate disease (n = 202) and fracture (n = 202) clinics in an urban Canadian teaching hospital (Sharpe study), and people with HIV/AIDS at 10 specialty care clinics and three primary care practices affiliated with the HIV Ontario Observational Database (n = 431). VARIABLES AND OUTCOME MEASURES: The survey instruments included questions about opinion of label, role in treatment decision-making (the Problem Solving Decision Making scale), trust, use of information and health status. RESULTS: Our respondents moderately liked the label 'patient'. The other alternatives evoked moderate to strong dislike. CONCLUSIONS: Many alternatives to 'patient' incorporate assumptions (e.g. a market relationship) which care recipients may also find objectionable. People who are receiving care find the label 'patient' much less objectionable than the alternatives that have been suggested. PMID- 16266423 TI - Consumer-driven health care: building partnerships in research. AB - Over the past four decades, there has been a widespread movement to increase the involvement of patients and the public in health care. Strategies to effectively foster consumer participation are occurring within all research activities from research priority setting to utilization. One of the ten principles of the Cochrane Collaboration is to 'enable wide participation', and this includes consumers. The Cochrane Musculoskeletal Group (CMSG) is a review group of 50 within the Collaboration that has been working to increase consumer participation since its inception in 1993. Based in Canada, the CMSG has embraced the concept of knowledge translation as advocated by the Canadian Institutes of Health Research. The emphasis in knowledge translation is on interactions or partnerships between researchers and users to facilitate the use of relevant research in decision making. While the CMSG recognizes the importance of reaching all users, much of its work has focused on developing relationships with people with musculoskeletal diseases to enhance consumer participation in research. The CMSG has built a network of consumer members who guide research priorities, peer review systematic reviews and also promote and facilitate consumer-appropriate knowledge dissemination. Consumers were recruited through links with other arthritis organizations and the recruitment continues. Specific roles were established for the consumer team and responsibilities of the CMSG staff developed. The continuing development of a diversified team of consumer participants enables the CMSG to produce and promote access to high quality relevant systematic reviews and summaries of those reviews to the consumer. PMID- 16266424 TI - The teamwork file: National Cancer Alliance 2003. PMID- 16266429 TI - Single subject pharmacological-MRI (phMRI) study: modulation of brain activity of psoriatic arthritis pain by cyclooxygenase-2 inhibitor. AB - We use fMRI to examine brain activity for pain elicited by palpating joints in a single patient suffering from psoriatic arthritis. Changes in these responses are documented when the patient ingested a single dose of a selective cyclooxygenase 2 inhibitor (COX-2i). We show that mechanical stimulation of the painful joints exhibited a cortical activity pattern similar to that reported for acute pain, with activity primarily localized to the thalamus, insular, primary and secondary somatosensory cortices and the mid anterior cingulum. COX-2i resulted in significant decreased in reported pain intensity and in brain activity after 1 hour of administration. The anterior insula and SII correlated with pain intensity, however no central activation site for the drug was detected. We demonstrate the similarity of the activation pattern for palpating painful joints to brain activity in normal subjects in response to thermal painful stimuli, by performing a spatial conjunction analysis between these maps, where overlap is observed in the insula, thalamus, secondary somatosensory cortex, and anterior cingulate. The results demonstrate that one can study effects of pharmacological manipulations in a single subject where the brain activity for a clinical condition is delineated and its modulation by COX-2i demonstrated. This approach may have diagnostic and prognostic utility. PMID- 16266430 TI - The chromosomes of the Filariae. AB - An understanding of the nature of the chromosomes of the filariae is expected to greatly assist the future interpretation of genome data. Filarial development is not eutelic, and there does not seem to be a fixed number of cell divisions in the way that there is in Caenorhabditis. It is not clear whether the chromosomes of the filariae have localized centromeres or whether they are holocentric. Sex determination is by a chromosomal "balance" X0 system in most filariae, but in some Onchocercidae there has been a chromosomal fusion to create a neo-XY system. It is presumed that the molecular basis of sex determination in filariae is similar to Caenorhabditis. The ancestral karyotype of the filariae is probably 5A+X0, but in some Onchocercidae this has been reduced to 4A+XY, and in O. volvulus and O. gibsoni it has been further reduced to 3A+XY. Onchocerca volvulus and O. gibsoni both have supernumary (B-) chromosomes and in O. volvulus there is a single active nucleolus organising region near the middle of the long autosome. PMID- 16266428 TI - The crucial role of particle surface reactivity in respirable quartz-induced reactive oxygen/nitrogen species formation and APE/Ref-1 induction in rat lung. AB - Persistent inflammation and associated excessive oxidative stress have been crucially implicated in quartz-induced pulmonary diseases, including fibrosis and cancer. We have investigated the significance of the particle surface reactivity of respirable quartz dust in relation to the in vivo generation of reactive oxygen and nitrogen species (ROS/RNS) and the associated induction of oxidative stress responses in the lung. Therefore, rats were intratracheally instilled with 2 mg quartz (DQ12) or quartz whose surface was modified by either polyvinylpyridine-N-oxide (PVNO) or aluminium lactate (AL). Seven days after instillation, the bronchoalveolar lavage fluid (BALF) was analysed for markers of inflammation (total/differential cell counts), levels of pulmonary oxidants (H2O2, nitrite), antioxidant status (trolox equivalent antioxidant capacity), as well as for markers of lung tissue damage, e.g. total protein, lactate dehydrogenase and alkaline phosphatase. Lung homogenates as well as sections were investigated regarding the induction of the oxidative DNA-lesion/oxidative stress marker 8-hydroxy-2'-deoxyguanosine (8-OHdG) using HPLC/ECD analysis and immunohistochemistry, respectively. Homogenates and sections were also investigated for the expression of the bifunctional apurinic/apyrimidinic endonuclease/redox factor-1 (APE/Ref-1) by Western blotting and immunohistochemistry. Significantly increased levels of H2O2 and nitrite were observed in rats treated with non-coated quartz, when compared to rats that were treated with either saline or the surface-modified quartz preparations. In the BALF, there was a strong correlation between the number of macrophages and ROS, as well as total cells and RNS. Although enhanced oxidant generation in non coated DQ12-treated rats was paralleled with an increased total antioxidant capacity in the BALF, these animals also showed significantly enhanced lung tissue damage. Remarkably however, elevated ROS levels were not associated with an increase in 8-OHdG, whereas the lung tissue expression of APE/Ref-1 protein was clearly up-regulated. The present data provide further in vivo evidence for the crucial role of particle surface properties in quartz dust-induced ROS/RNS generation by recruited inflammatory phagocytes. Our results also demonstrate that quartz dust can fail to show steady-state enhanced oxidative DNA damage in the respiratory tract, in conditions were it elicits a marked and persistent inflammation with associated generation of ROS/RNS, and indicate that this may relate to compensatory induction of APE/Ref-1 mediated base excision repair. PMID- 16266432 TI - Proposed methods for testing and selecting the ERCC external RNA controls. AB - The External RNA Control Consortium (ERCC) is an ad-hoc group with approximately 70 members from private, public, and academic organizations. The group is developing a set of external RNA control transcripts that can be used to assess technical performance in gene expression assays. The ERCC is now initiating the Testing Phase of the project, during which candidate external RNA controls will be evaluated in both microarray and QRT-PCR gene expression platforms. This document describes the proposed experiments and informatics process that will be followed to test and qualify individual controls. The ERCC is distributing this description of the proposed testing process in an effort to gain consensus and to encourage feedback from the scientific community. On October 4-5, 2005, the ERCC met to further review the document, clarify ambiguities, and plan next steps. A summary of this meeting and changes to the test plan are provided as an appendix to this manuscript. PMID- 16266433 TI - Apical membrane P2Y4 purinergic receptor controls K+ secretion by strial marginal cell epithelium. AB - BACKGROUND: It was previously shown that K+ secretion by strial marginal cell epithelium is under the control of G-protein coupled receptors of the P2Y family in the apical membrane. Receptor activation by uracil nucleotides (P2Y2, P2Y4 or P2Y6) leads to a decrease in the electrogenic K+ secretion. The present study was conducted to determine the subtype of the functional purinergic receptor in gerbil stria vascularis, to test if receptor activation leads to elevation of intracellular [Ca2+] and to test if the response to these receptors undergoes desensitization. RESULTS: The transepithelial short circuit current (Isc) represents electrogenic K+ secretion and was found to be decreased by uridine 5' triphosphate (UTP), adenosine 5'-triphosphate (ATP) and diadenosine tetraphosphate (Ap4A) but not uridine 5'-diphosphate (UDP) at the apical membrane of marginal cells of the gerbil stria vascularis. The potencies of these agonists were consistent with rodent P2Y4 and P2Y2 but not P2Y6 receptors. Activation caused a biphasic increase in intracellular [Ca2+] that could be partially blocked by 2-aminoethoxy-diphenyl borate (2-APB), an inhibitor of the IP3 receptor and store-operated channels. Suramin (100 microM) did not inhibit the effect of UTP (1 microM). The ineffectiveness of suramin at the concentration used was consistent with P2Y4 but not P2Y2. Transcripts for both P2Y2 and P2Y4 were found in the stria vascularis. Sustained exposure to ATP or UTP for 15 min caused a depression of Isc that appeared to have two components but with apparently no chronic desensitization. CONCLUSION: The results support the conclusion that regulation of K+ secretion across strial marginal cell epithelium occurs by P2Y4 receptors at the apical membrane. The apparent lack of desensitization of the response is consistent with two processes: a rapid-onset phosphorylation of KCNE1 channel subunit and a slower-onset of regulation by depletion of plasma membrane PIP2. PMID- 16266434 TI - Analysis of four DLX homeobox genes in autistic probands. AB - BACKGROUND: Linkage studies in autism have identified susceptibility loci on chromosomes 2q and 7q, regions containing the DLX1/2 and DLX5/6 bigene clusters. The DLX genes encode homeodomain transcription factors that control craniofacial patterning and differentiation and survival of forebrain inhibitory neurons. We investigated the role that sequence variants in DLX genes play in autism by in depth resequencing of these genes in 161 autism probands from the AGRE collection. RESULTS: Sequencing of exons, exon/intron boundaries and known enhancers of DLX1, 2, 5 and 6 identified several nonsynonymous variants in DLX2 and DLX5 and a variant in a DLX5/6 intragenic enhancer. The nonsynonymous variants were detected in 4 of 95 families from which samples were sequenced. Two of these four SNPs were not observed in 378 undiagnosed samples from North American populations, while the remaining 2 were seen in one sample each. CONCLUSION: Segregation of these variants in pedigrees did not generally support a contribution to autism susceptibility by these genes, although functional analyses may provide insight into the biological understanding of these important proteins. PMID- 16266435 TI - Absence of force suppression in rabbit bladder correlates with low expression of heat shock protein 20. AB - BACKGROUND: Nitroglycerin can induce relaxation of swine carotid artery without sustained reductions in [Ca2+]i or myosin regulatory light chain (MRLC) phosphorylation. This has been termed force suppression and been found to correlate with ser16-phosphorylation of heat shock protein 20 (HSP20). We tested for the existence of this mechanism in a smooth muscle that is not responsive to nitric oxide. METHODS: Isometrically mounted mucosa free rabbit bladder strips were contracted with carbachol and relaxed with 8-Br-cGMP, forskolin, or isoprenaline. RESULTS: Contraction was associated with a highly cooperative relation between MRLC phosphorylation and force such that very small increases in MRLC phosphorylation induced large increases in force. Relaxation induced by 8-Br cGMP, forskolin, or isoprenaline did not shift the MRLC phosphorylation-force relation from that observed with carbachol alone, i.e. there was no force suppression. HSP20 content was negligible (approximately two hundred-fold less than swine carotid). CONCLUSION: The lack of force suppression in the absence of HSP20 is consistent with the hypothesized role for HSP20 in the force suppression observed in tonic smooth muscles. PMID- 16266437 TI - ATP-sensitive potassium channel (KATP channel) expression in the normal canine pancreas and in canine insulinomas. AB - BACKGROUND: Pancreatic beta cells express ATP-sensitive potassium (KATP) channels that are needed for normal insulin secretion and are targets for drugs that modulate insulin secretion. The KATP channel is composed of two subunits: a sulfonylurea receptor (SUR 1) and an inward rectifying potassium channel (Kir6.2). KATP channel activity is influenced by the metabolic state of the cell and initiates the ionic events that precede insulin exocytosis. Although drugs that target the KATP channel have the expected effects on insulin secretion in dogs, little is known about molecular aspects of this potassium channel. To learn more about canine beta cell KATP channels, we studied KATP channel expression by the normal canine pancreas and by insulin-secreting tumors of dogs. RESULTS: Pancreatic tissue from normal dogs and tumor tissue from three dogs with histologically-confirmed insulinomas was examined for expression of KATP channel subunits (SUR1 and Kir6.2) using RT-PCR. Normal canine pancreas expressed SUR1 and Kir6.2 subunits of the KATP channel. The partial nucleotide sequences for SUR1 and Kir6.2 obtained from the normal pancreas showed a high degree of homology to published sequences for other mammalian species. SUR1 and Kir6.2 expression was observed in each of the three canine insulinomas examined. Comparison of short sequences from insulinomas with those obtained from normal pancreas did not reveal any mutations in either SUR1 or Kir6.2 in any of the insulinomas. CONCLUSION: Canine pancreatic KATP channels have the same subunit composition as those found in the endocrine pancreases of humans, rats, and mice, suggesting that the canine channel is regulated in a similar fashion as in other species. SUR1 and Kir6.2 expression was found in the three insulinomas examined indicating that unregulated insulin secretion by these tumors does not result from failure to express one or both KATP channel subunits. PMID- 16266436 TI - Cerebrospinal fluid HIV infection and pleocytosis: relation to systemic infection and antiretroviral treatment. AB - BACKGROUND: Central nervous system (CNS) exposure to HIV is a universal facet of systemic infection. Because of its proximity to and shared barriers with the brain, cerebrospinal fluid (CSF) provides a useful window into and model of human CNS HIV infection. METHODS: Prospective study of the relationships of CSF to plasma HIV RNA, and the effects of: 1) progression of systemic infection, 2) CSF white blood cell (WBC) count, 3) antiretroviral therapy (ART), and 4) neurological performance. One hundred HIV-infected subjects were cross sectionally studied, and 28 were followed longitudinally after initiating or changing ART. RESULTS: In cross-sectional analysis, HIV RNA levels were lower in CSF than plasma (median difference 1.30 log10 copies/mL). CSF HIV viral loads (VLs) correlated strongly with plasma VLs and CSF WBC counts. Higher CSF WBC counts associated with smaller differences between plasma and CSF HIV VL. CSF VL did not correlate with blood CD4 count, but CD4 counts <50 cells/microL associated with a low prevalence of CSF pleocytosis and large differences between plasma and CSF VL. CSF HIV RNA correlated neither with the severity of the AIDS dementia complex (ADC) nor abnormal quantitative neurological performance, although these measures were associated with depression of CD4 counts. In subjects starting ART, those with lower CD4 counts had slower initial viral decay in CSF than in plasma. In all subjects, including five with persistent plasma viremia and four with new-onset ADC, CSF HIV eventually approached or reached the limit of viral detection and CSF pleocytosis resolved. CONCLUSION: CSF HIV infection is common across the spectrum of infection and is directly related to CSF pleocytosis, though whether the latter is a response to or a contributing cause of CSF infection remains uncertain. Slowing in the rate of CSF response to ART compared to plasma as CD4 counts decline indicates a changing character of CSF infection with systemic immunological progression. Longer-term responses indicate that CSF infection generally responds well to ART, even in the face of systemic virological failure due to drug resistance. We present simple models to explain the differing relationships of CSF to plasma HIV in these settings. PMID- 16266438 TI - The trend of susceptibilities to amphotericin B and fluconazole of Candida species from 1999 to 2002 in Taiwan. AB - BACKGROUND: Candida species have various degrees of susceptibility to common antifungal drugs. The extent of resistance to amphotericin B and fluconazole of Candida glabrata isolates causing candidemia has been reported. Active surveillance may help us to monitor the trend of susceptibility to antifungal drugs and to determine if there is an emerging co-resistance to both drugs of Candida species, specifically, of C. glabrata in Taiwan. METHODS: The susceptibilities to amphotericin B and fluconazole of Candida species collected in 1999 and 2002 of the Taiwan Surveillance of Antimicrobial Resistance of Yeasts (TSARY) were determined by the microdilution method. RESULTS: The antifungal susceptibilities of 342 and 456 isolates collected from 11 hospitals participating in both TSARY 1999 and TSARY 2002, respectively, have been determined. The resistance rate to amphotericin B has increased from 0.3% in the TSARY1999 to 2.2% in the TSARY 2002. In contrast, the resistance rate to fluconazole has decreased from 8.8% to 2.2%. Nevertheless, significantly more C. glabrata isolates were not susceptible to fluconazole in the TSARY 2002 (47.4%) than that in the TSARY 1999 (20.8%). There were 9.8% and 11% of C. glabrata isolates having susceptible-dose dependent and resistant phenotype to fluconazole in the TSARY 1999, verse 45.3% and 2.1% in the TSARY 2002. CONCLUSION: There was an increase of resistance rate to amphotericin B in C. glabrata. On the other hand, although the resistance rate to fluconazole has decreased, almost half of C. glabrata isolates were not susceptible to this drug. Hence, continuous monitoring the emerging of co-resistance to both amphotericin B and fluconazole of Candida species, specifically, of C. glabrata, will be an important early warning system. PMID- 16266439 TI - The gene for a lectin-like protein is transcriptionally activated during sexual development, but is not essential for fruiting body formation in the filamentous fungus Sordaria macrospora. AB - BACKGROUND: The filamentous fungus Sordaria macrospora forms complex three dimensional fruiting bodies called perithecia that protect the developing ascospores and ensure their proper discharge. In previous microarray analyses, several genes have been identified that are downregulated in sterile mutants compared to the wild type. Among these genes was tap1 (transcript associated with perithecial development), a gene encoding a putative lectin homolog. RESULTS: Analysis of tap1 transcript levels in the wild type under conditions allowing only vegetative growth compared to conditions that lead to fruiting body development showed that tap1 is not only downregulated in developmental mutants but is also upregulated in the wild type during fruiting body development. We have cloned and sequenced a 3.2 kb fragment of genomic DNA containing the tap1 open reading frame and adjoining sequences. The genomic region comprising tap1 is syntenic to its homologous region in the closely related filamentous fungus Neurospora crassa. To determine whether tap1 is involved in fruiting body development in S. macrospora, a knockout construct was generated in which the tap1 open reading frame was replaced by the hygromycin B resistance gene hph under the control of fungal regulatory regions. Transformation of the S. macrospora wild type with this construct resulted in a tap1 deletion strain where tap1 had been replaced by the hph cassette. The knockout strain displayed no phenotypic differences under conditions of vegetative growth and sexual development when compared to the wild type. Double mutants carrying the Deltatap1 allele in several developmental mutant backgrounds were phenotypically similar to the corresponding developmental mutant strains. CONCLUSION: The tap1 transcript is strongly upregulated during sexual development in S. macrospora; however, analysis of a tap1 knockout strain shows that tap1 is not essential for fruiting body formation in S. macrospora. PMID- 16266441 TI - Anthropological insights into the use of race/ethnicity to explore genetic contributions to disparities in health. AB - Anthropological insights into the use of race/ethnicity to explore genetic contributions to disparities in health were developed using in-depth qualitative interviews with editorial staff from nineteen genetics journals, focusing on the methodological and conceptual mechanisms required to make race/ethnicity a genetic variable. As such, these analyses explore how and why race/ethnicity comes to be used in the context of genetic research, set against the background of continuing critiques from anthropology and related human sciences that focus on the social construction, structural correlates and limited genetic validity of racial/ethnic categories. The analyses demonstrate how these critiques have failed to engage geneticists, and how geneticists use a range of essentially cultural devices to protect and separate their use of race/ethnicity as a genetic construct from its use as a societal and social science resource. Given its multidisciplinary, biosocial nature and the cultural gaze of its ethnographic methodologies, anthropology is well placed to explore the cultural separation of science and society, and of natural and social science disciplines. Anthropological insights into the use of race/ethnicity to explore disparities in health suggest that moving beyond genetic explanations of innate difference might benefit from a more even-handed critique of how both the natural and social sciences tend to essentialize selective elements of race/ethnicity. Drawing on the example of HIV/AIDS, this paper demonstrates how public health has been undermined by the use of race/ethnicity as an analytical variable, both as a cipher for innate genetic differences in susceptibility and response to treatment, and in its use to identify 'core groups' at greater risk of becoming infected and infecting others. Clearly, a tendency for biological reductionism can place many biomedical issues beyond the scope of public health interventions, while socio-cultural essentialization has tended to stigmatize 'unhealthy behaviours' and the communities where these are more prevalent. PMID- 16266440 TI - Homeopathic medical practice: long-term results of a cohort study with 3981 patients. AB - BACKGROUND: On the range of diagnoses, course of treatment, and long-term outcome in patients who chose to receive homeopathic medical treatment very little is known. We investigated homeopathic practice in an industrialized country under everyday conditions. METHODS: In a prospective, multicentre cohort study with 103 primary care practices with additional specialisation in homeopathy in Germany and Switzerland, data from all patients (age > 1 year) consulting the physician for the first time were observed. The main outcome measures were: Patient and physician assessments (numeric rating scales from 0 to 10) and quality of life at baseline, and after 3, 12, and 24 months. RESULTS: A total of 3,981 patients were studied including 2,851 adults (29% men, mean age 42.5 +/- 13.1 years; 71% women, 39.9 +/- 12.4 years) and 1,130 children (52% boys, 6.5 +/- 3.9 years; 48% girls, 7.0 +/- 4.3 years). Ninety-seven percent of all diagnoses were chronic with an average duration of 8.8 +/- 8 years. The most frequent diagnoses were allergic rhinitis in men, headache in women, and atopic dermatitis in children. Disease severity decreased significantly (p < 0.001) between baseline and 24 months (adults from 6.2 +/- 1.7 to 3.0 +/- 2.2; children from 6.1 +/- 1.8 to 2.2 +/- 1.9). Physicians' assessments yielded similar results. For adults and young children, major improvements were observed for quality of life, whereas no changes were seen in adolescents. Younger age and more severe disease at baseline were factors predictive of better therapeutic success. CONCLUSION: Disease severity and quality of life demonstrated marked and sustained improvements following homeopathic treatment period. Our findings indicate that homeopathic medical therapy may play a beneficial role in the long-term care of patients with chronic diseases. PMID- 16266442 TI - Urban health and pharmaceutical consumption in Delhi, India. AB - This paper interrogates the routine and unproblematic use of terms such as 'self medication' in biomedical and anthropological discourse. A typical depiction of the social factors that explain the practice of 'self-medication' in India is to put together the supply side factors (such as protection offered by the government for the production of generic drugs, especially in the small scale sector, and expansion of the number of drug store outlets), with the increasing demand for allopathic drugs. The paper provides an ethnographic account of the intricate connections between households and biomedical practitioners in urban neighbourhoods in Delhi. It breaks away from the conventional opposition drawn between the practices of physicians and the beliefs of their patients, and suggests that what constitutes the medical environments of these neighbourhoods is the product of medical practices, household economies and concepts of disease. Thus pharmaceutical use is determined as much by practices of dispensation and by how practitioners understand what constitutes therapy as by household understanding of the normal and the pathological. This paper uses both quantitative data and narrative interviews to provide an in-depth understanding of the circulation of pharmaceuticals within the life worlds of the urban poor. PMID- 16266444 TI - Abstain or die: the development of HIV/AIDS policy in Botswana. AB - This paper traces the development of policies dealing with HIV/AIDS in Botswana from their beginning in the late 1980s to the current programme to provide population-wide anti-retroviral therapy (ARV). Using a variety of source material, including long-term ethnographic research, it seeks to account for the failure of Western-inspired approaches in dealing with the pandemic. It does this by looking at the cultural and institutional features that have created resistance to the message and inhibited effective implementation. The negative response to the first educational campaign stressing condom use is described and contextualized in terms of Tswana ideas of morality and illness. Nor, as was initially expected, did the introduction of free ARV therapy operate to break the silence and stigma that had developed around the disease. Take-up was very slow, and did not operate to encourage widespread testing. In 2003, key policymakers in Botswana began to argue for a break with the AIDS 'exceptionalism' position, with its emphasis on voluntarism, confidentiality and the human rights of patients. This resulted in routine testing being introduced in 2004. This links to a major argument running through the paper which is that the failure of policy cannot be attributed solely to the nature of local populations. Western cultural assumptions about 'good practice' also require critical examination. PMID- 16266443 TI - Public engagement with science? Local understandings of a vaccine trial in the Gambia. AB - This paper considers how parents engage with a large, internationally supported childhood pneumococcal vaccine trial in The Gambia. Current analysis and professional reflection on public engagement is strongly shaped by the imperatives of public health and research institutions, and is thus couched in terms of acceptance and refusal, and 'informed consent'. In contrast Gambian parents' perspectives on the trial are couched in conceptual and experiential terms that are linked to their wider dilemmas of raising infants amidst the hazards of globally connected village life. Ethnographic research reveals how for most parents, longer-term experiences of the organization managing the research (the UK Medical Research Council Laboratories in The Gambia) as a health providing institution override their reflection on trial-specific aims. A decision to participate in the trial involves a perceived balance of benefit and danger -- in the extreme, of free medical treatment, versus one's child being drained of blood for sale to Europe. Social relations (especially gender relations) shape this calculus and study participation. This case suggests how the idea of 'public engagement with science' in a globalized context might be recast, with implications for debates in biomedical ethics, and the sustainability of public participation in medical research. PMID- 16266445 TI - The grant is what I eat: the politics of social security and disability in the post-apartheid South African state. AB - In South Africa, disability grant allocation has been under review and tensions are evident in government rhetoric stressing welfare provision on the one hand, and encouraging 'rationalization' on the other. This ambiguity is traced down to the level of grant negotiations between doctors and 'clients' in a psychiatry clinic in Khayelitsha. Here 'having nerves' embodies the distress associated with harsh circumstances and is deemed by supplicants as sufficient to secure a grant. The paper illustrates how national discourses influence the presentation and experience of suffering and the way in which doctors mediate diagnoses. The implications of local understandings of 'health citizenship' for expectations of the post-apartheid state are explored. PMID- 16266446 TI - Perceived nuisance of mosquitoes on the Isle of Sheppey, Kent, UK. AB - Little is known about the biting nuisance of mosquitoes in the UK, despite the high numbers found in some locations. A telephone questionnaire survey was used to determine the perceived nuisance of biting insects on the Isle of Sheppey, Kent, a place notorious for mosquitoes. Two hundred randomly selected individuals were interviewed and asked if they suffered from mosquito bites. If they answered yes, they were asked to describe where and when they were bitten, and what measures they took against mosquitoes. Forty-six percent of respondents completed the questionnaire. Of those, 50% reported being bitten by mosquitoes, mostly outside during the summer. Seventy percent said that most biting occurred during the evening and night. Of those respondents who protected themselves against biting (27), most used repellents (70%), with the remainder changing their behaviour to avoid mosquitoes, including closing or screening windows (33%), wearing thicker clothes (7%) and spraying insecticide (4%). One person slept under a bednet in summer (4%). This study provides evidence that on the Isle of Sheppey mosquitoes are considered a major nuisance by a sizeable proportion of the population. Since there is growing interest in the threat posed by new and emerging diseases in the UK, health authorities will need to make substantial efforts to inform and reassure the public about the threats posed by mosquitoes in areas where they are common. PMID- 16266447 TI - Anthropology, DOTS and understanding tuberculosis control in Nepal. AB - This paper argues for the inclusion of ethnography as a research methodology for understanding the effects of public health policy. To do this, the implementation of DOTS (Directly Observed Therapy, Short-course) -- the World Health Organization (WHO) prescribed policy for the control of the infectious disease tuberculosis -- is explored in the context of Nepal. A brief history of DOTS and its implementation in Nepal is outlined, and the way it has been represented by those within the Nepal Tuberculosis Programme (NTP) is described. This is followed by an outline of the research done in relation to this, and the ethnographic methods used. These ethnographic data are then interpreted and analysed in relation to two specific areas of concern. Firstly, the effects around the epidemiological uses of 'cases' is explored; it is argued that a tightening of the definitional categories so necessary for the programme to be stabilized for comparative purposes has profound material effects in marginalizing some from treatment. Secondly, the paper examines some of the implications and effects relating to the way that the 'directly observed' component was implemented. The discussion explores how current debate on DOTS has been played out in some medical journals. It argues for the importance of ethnography as a method for understanding certain questions that cannot be answered by particular, and increasingly dominant, research ideologies informed by randomized controlled trials. This raises important issues about the nature of 'evidence' in debates on the relationship of research to policy. PMID- 16266448 TI - [The important value of ECG in evaluating myocardial perfusion]. PMID- 16266449 TI - [Effect of Chinese guidelines issued in 2001 on in-hospital management and prognosis of patients with acute myocardial infarction]. AB - OBJECTIVE: To evaluate the effect of Chinese guidelines issued on December 2001 on in-hospital management and prognosis of patients with acute myocardial infarction. METHODS: A retrospective study was carried out in patients hospitalized in our hospital with acute myocardial infarction from January 1994 to December 2004. RESULTS: There were 1783 patients enrolled in our study. Reperfusion therapy was undergone in 21.7% of patients hospitalized between 1994 and 2001, and in 35.8% of patients hospitalized between 2002 and 2004 (P < 0.001). Beta-blockers, ACE inhibitors and/or angiotensin receptor blockers, lipid regulating agents and antithrombins were used more extensively between 2002 and 2004 than before (all P < 0.001). There were no significant differences in the usage of nitrates and antiplatelets before and after the guidelines was issued (98.8% vs 97.9%, P = 0.172; 97.4% vs 98.6%, P = 0.113 respectively). After the guidelines issued, the incidence of angina pectoris, heart failure and death in hospital were lower than before (32.2% vs 41.2%, P < 0.001; 17.2% vs 26.2%, P < 0.001; 6.4% vs 9.4%, P = 0.038). CONCLUSIONS: Chinese guidelines issued on December 2001 have great effect on the management and prognosis of patients with acute myocardial infarction. After the guidelines was issued the management became more standardized and the incidence of in-hospital complications was lower than before. PMID- 16266450 TI - [Study on early fibrinolytic therapy to avoid acute myocardial infarction]. AB - OBJECTIVE: To investigate the frequency of aborted AMI and clinical characteristics of the patients received prompt fibrinolytic therapy. METHODS: 1120 patients with AMI were divided into two groups, true AMI group and aborted AMI group. Aborted AMI was defined as maximal creatine kinase-MB < or = 2 x upper limit of normal coupled with the presence of resolution of chest pain and 50% of ST-segment deviation within 2 hours after onset of therapy. We compared some characteristic of two groups such as the fibrinolytic time after symptom onset and the frequency of aborted AMI. RESULTS: The reopening ratio of infarct was 80.5%. 7.1% of the patients escaped myocardial necrosis. Aborted AMI was highest frequency within the first hour (22.0%) than other time groups (P < 0.01); There were no significant differences in the frequency of Aborted AMI in UK group, SK group and rt-PA group (7.0%, 6.7%, 7.1%, P > 0.05); The rate of Killip III/IV, major arrhythmias, angina pectoris and mortality at 30 day in aborted AMI patients compared with those who had true AMI was 3.9% versus 17.1%, 18.0% versus 30.0%, 1.3% versus 8.0%, 0 versus 6.0%, respectively (P < 0.01). CONCLUSION: Prompt fibrinolytic therapy improved the likelihood of aborted AMI and clinical outcomes. The frequency of aborted AMI has no relationship with fibrinolytic drug, but closely related to the starting time of treatment from symptom onset. PMID- 16266452 TI - [Comparison of current clinical practice and guideline application in therapies of ACS: findings from the Multi-central Collaborative Group on Chinese registry of acute coronary events]. AB - OBJECTIVE: To evaluate the current practice of medications that have been proven effective by evidence-based medicine on patients with ACS. METHODS: (One thousand three hundred and one patients with ACS from 12 hospitals in China received different therapies in hospitals. We analyzed the status of application of reperfusion and aspirin, ACEI, beta-blocker, LWMH and cholesterol lowering agents on different ACS patients. RESULTS: (1) Percentage of medications in STEMI patients in different periods: aspirin (95.9%-100.0%), ACEI (72.0%-88.4%), beta blocker (62.7%-74.5%), LWMH (84.7%-100.0%), cholesterol lowering agents (72.5% 93.0%); (2) In NSTEMI patients: aspirin (100%), LWMH in NSTEMI patients (84.2% 100.0%), in UAP patients (65.1%-87.2%); (3) Among STEMI patients, 48.9% received primary PCI, 12% received Fibrinolytics only, and nearly 28.8% did not receive any form of reperfusion. (4) 50% of NSTEACS patients received PCI therapy. CONCLUSION: We conclude that in the developed regions in China the application of medications being proved effective by evidence-based medicine in clinical practice is better than that reported in other countries, and that the application status in China could be further improved. PMID- 16266451 TI - [The relationship between circulating endothelial progenitor cells and the risk factors of CHD as well as the severity of coronary lesions, and its clinical significance]. AB - OBJECTIVE: To investigate the correlation between circulating endothelial progenitor cells (EPCs) and the risk factors of coronary heart disease (CHD) as well as the severity of coronary lesions, and its clinical significance. METHODS: 42 patients with CHD and 36 patients excluding CHD (control) were studied. Total mononuclear cells were isolated from peripheral blood by Ficoll density gradient centrifugation, and were cultured in M199 medium supplemented with 20% fetal bovine serum, 50 ng/ml vascular endothelial growth factor (VEGF). After 14 days cultured, the numbers of colony-forming units of EPCs were counted by phase contrast microscope. The relationship between the number of colony-forming units of EPCs and the risk factors of CHD (such as age, gender, hypertension, hypercholesterolemia, diabetes, smoking, positive family history of CHD) as well as the severity of coronary lesions were assessed. RESULTS: The number of risk factors of CHD was significantly correlated with a reduction of EPCs levels (r = 0.436, P = 0.014). Smoking was associated with significantly lower EPCs levels, whereas a minor but nonsignificant reduction of EPCs levels was detected in the presence of gender, hypertension, and a positive family history of CHD. It was observed that low density lipoprotein (LDL) and uric acid were negatively correlated with the number of colony-forming units of circulating EPCs (P < 0.05). A correlation existed between age, high density lipoprotein, apoprotein A and levels of circulating EPCs, however, this relation was not statistically significant. The number of colony-forming units of circulating EPCs in CHD groups was significantly lower than those in control group (12.8 +/- 6.34 versus 37.0 +/ 5.5, P < 0.001); and the circulating EPCs level of coronary artery lesion group (including single, double, triple vessels disease) was significantly lower than that of control group (P < 0. 01). CONCLUSIONS: The level of circulating EPCs was inversely associated with the risk factor scores of CHD and the severity of coronary artery lesion. These finding imply that endothelial injury in the absence of sufficient circulating EPCs may affect the degree of the heart disorder and the clinical situation. PMID- 16266453 TI - [Therapeutic value of trimetazidine in patients with coronary heart disease and left ventricular dysfunction]. AB - OBJECTIVE: Trimetazidine has been shown to have anti-ischemic properties by improving exercise tolerance without haemodynamic effects and direct cyto protective effects on the myocardium. The aim of this study was to assess the effect of trimetazidine on the treatment of patients with ischemic cardiomyopathy, and to assess the anti-angina efficacy and the clinical tolerance of TMZ. METHODS: 60 ischemic cardiomyopathy patients on conventional therapy were enrolled in this open multiple center clinical trial and received TMZ (20 mg 3 times per day) for six months. Patients were followed up for six months and assessments included clinical evaluation, electrocardiography and echocardiography. RESULTS: All patients tolerated 60 mg of trimetazidine for six months, left ventricular ejection fraction (LVEF) was increased from 32.4 +/- 6.3% to 41.8 +/- 10.8% (P < 0.0001). Stage of NYHA classification was improved in most patients (P < 0.0001). Attack of angina was relieved and consumption of nitroglycerin reduced (both P < 0.001). CONCLUSION: Trimetazidine seems to improve clinical status and ejection fraction, and reduce angina onsets without adverse effects in patients with ischemic cardiomyopathy. PMID- 16266454 TI - [Comparable analysis of in-hospital mortality in patients with acute myocardial infarction]. AB - OBJECTIVE: To investigate the change of the mortality of AMI and influence factors within 20 years. METHODS: Clinic data of 134 AMI patients from 1980 to 1983, 354 AMI patients from 1990 to 1993 and 817 AMI patients from 2000 to 2003 were comparably analyzed. RESULTS: In hospital mortality of AMI was 22.4% from 1980 to 1983, 14.4% from 1990 to 1993 and 9.2% from 2000 to 2003, respectively (P < 0.01). The decrease of in-hospital mortality in male was more significant than in female (P < 0.01). The corresponding factors for decrease of mortality were younger than 60 years old, first onset of AMI, successful rescue of cardiac arrest and reperfusion management of infarction relative artery. The disadvantage factor was female. CONCLUSIONS: Improvement of medical and reperfusion management of AMI conduced in significant decreases of hospital mortality. PMID- 16266455 TI - [Noninvasive assessment of left anterior descending and right coronary artery flow reserve by transthoracic Doppler echocardiography in patients with coronary artery disease]. AB - OBJECTIVE: To assess the feasibility of evaluation of CFR for LAD and right coronary artery (RCA) as well as diagnostic accuracy for patency of each vessel by transthoracic Doppler echocardiography (TTDE). METHODS: 65 consecutive patients (age 58 +/- 14, ejection fraction 49% +/- 8%, 48 men and 17 women) were studied for CFR by TTDE with adenosine infusion (140 microg.kg(-1).min(-1)). LAD flow was interrogated nearby the apex in modified 2-ch apical view. RCA flow was interrogated at mid posterior wall on distal part of the posterior descending branch (PDA) in modified 2-ch apical view for PDA. RESULTS: Flow for LAD was detected in all patients (4 with use of contrast agent) and for RCA in 55 patients (5 with contrast agent). Two patients had an occluded RCA. Feasibility of RCA flow detection was 55/63 (87%). In all patients maximal flow of each branch was detected in less than 2 min of adenosine infusion (140 mg.kg(-1).min( 1)). Total time for both CFR estimation was 7.3 +/- 1.6 min. Selecting 2.0 or 1.8 as CFR cut-off value for diagnosis of coronary stenosis (> or = 50% in diameter) or severe stenosis (> or = 70%) was found fitted for both LAD and RCA with similar diagnostic performance (sensitivity 89%/93%, specificity 86%/84%, area under curve 0.89/0.92 respectively, in stenosis diagnosis and sensitivity 88%/89%, specificity 90%/83%, area under curve 0.94/0.95 respectively in severe stenosis diagnosis). CONCLUSIONS: CFR evaluation of both distal LAD and PDA is of high feasibility and can provide high diagnostic yield for relevant vessel patency. PMID- 16266456 TI - [Efficacy and safety of the coronary intervention therapy to prevent electrical storm in patients with coronary artery disease but without myocardial infarction]. AB - OBJECTIVE: To study the efficacy and safety of the coronary interventional therapy to prevent electrical storm (ES) in patients with coronary artery diseases (CAD) but without myocardial infarction or ischemic cardiomyopathy. METHODS: Coronary angiography and stent implantation were performed in CAD patients with ES as major symptom, according to the standardized methods. Holter Electrocardiography was recorded regularly during follow-up. RESULTS: Six patients, five male and one female, with mean age of 49.5 +/- 9.1 year-old, were hospitalized. In 2 patients with repetitive syncope, multiple episodes of ventricular tachycardia and/or ventricular fibrillation (VF) were documented by Holter recording. One patient developed VF during exercise test. Three patients experienced chest pain and multiple episodes of cardiac arrest. Before procedure, averaged 16.5 +/- 5.3 episodes of syncope or VF were documented in 6 patients. Coronary angiography revealed severe one or multi-vessel diseases. Total 8 stents, including 3 drug-eluting stents, were implanted in 6 patients. Symptom, ST-T changes and ES disappeared after coronary stenting. During 4 month to 6.5 year follow-up (mean 47.7 +/- 30.7 months), ES was not documented, no appropriated shock occurred in patients with implantable defibrillator for 6.5 years. CONCLUSION: In CAD patients without myocardial infarction, coronary stenting can relieve the ischemic substrate of ES, hence prevents sudden death effectively. PMID- 16266457 TI - [The effect of homocysteine on fibrinolytic system in human umbilical vein endothelial cells]. AB - OBJECTIVES: To investigate the relationship between homocysteine (Hcy) and the fibrinolytic system in acute myocardial infarction (AMI) and human umbilical vein endothelial cells (HUVEC). METHODS: Cultured HUVEC was divided into 10 groups (0, 10, 50, 200, 500 micromol/L Hcy with or without 15 micromol/L of folic acid). There were 53 patients of acute myocardial infarction (AMI) and 48 healthy controls. The plasminogen activator inhibitor-1 (PAI-1) and activator of plasminogen (tPA) antigen levels in HUVEC's supernatant and plasma were measured with Elisa kit. Concentration of plasma Hcy was measured by reverse-phase high performance liquid chromatography with precolumn derivatization and fluorometric detection in the patients and healthy controls. Total RNA was extracted using the guanidinium isothiocyanate method. The semi-quantification of PAI-1 and tPA mRNA in HUVEC was carried out by reverse transcriptase-polymerase chain reaction (RT PCR). RESULTS: (1) PAI-1 mRNA and secreted protein levels were both significantly enhanced by Hcy at the concentration of 500 micromol/L, compared with the control group (P < 0.05). (2) The tPA mRNA and antigen levels were decreased significantly at concentration of 500 micromol/L of Hcy, compared with that of 10 micromol/L Hcy (P < 0.05), but compared with the control group (0 micromol/L), the tPA mRNA and antigen levels of 10 micromol/L of Hcy were much higher (P < 0.05). (3) The addition of folic acid reduced PAI-1 but increased tPA at both mRNA and protein levels, which were both obvious at concentrations of 500 micromol/L Hcy, compared with only Hcy group (P < 0.05). (4) Hcy, tPA, and PAI-1 antigen levels were increased in AMI group. Hcy is a independent risk factor of AMI (P < 0.05). There weren't significant correlation between Hcy and tPA or Hcy and PAI-1 in both groups (P > 0.05), although the coefficient correlation was higher in patients than in controls. CONCLUSIONS: These results suggested that hyperhomo-cysteinemia increased the incidence of thrombotic disease, which may be caused by decreasing the activity of fibrinolytic system, whereas, folic acid may be protective against the toxic action of Hcy. PMID- 16266458 TI - [The AGT genotype affects the antihypertensive effects of benazepril]. AB - OBJECTIVE: To investigate the association between M235T variant of angiotensinogen (AGT) gene and the blood pressure response to benazepril in a hypertensive cohort. METHODS: Benazepril (10-20 mg/day) was administered for 6 weeks to 251 essential hypertensives. Polymerase chain reaction (PCR) combined with restriction enzyme digestion was used to detect the polymorphism and the patients were classified as MM, MT or TT genotype. The changes in systolic and diastolic blood pressure (SBP and DBP) were analyzed for association with genotypes at the AGT gene locus. RESULTS: The MM genotype was observed in 23 patients (9.2%), the MT genotype in 104 patients (41.4%) and the TT genotype in 124 patients (49.4%). There was no association between these polymorphisms and the blood pressure responses in the total 251 patients. But based on the analysis stratified by age, the association between these polymorphism and the DBP responses was found in the old patients (> or = 60 years old) subgroup, the reduction in DBP was significantly greater in patients carrying the MM compared to MT or TT genotypes (14.8 +/- 4.8 mm Hg vs. 7.9 +/- 7.7 mm Hg or 9.8 +/- 6.4 mm Hg respectively; ANOVA, P = 0.034). CONCLUSION: The M235T polymorphism of the AGT gene was shown to influence the responses to benazepril in old hypertensive patients (> or = 60 years old). Thus, specific genotypes might predict the response to specific antihypertensive treatment. PMID- 16266459 TI - [A preliminary study on the role of anti-atrial fibrillation pacemaker in the prevention of atrial tachyarrhythmia]. AB - OBJECTIVE: To investigate the effect of anti-atrial fibrillation of Philos DDDR pacemaker on atrial tachyarrhythmia. METHODS: Thirty-eight patients with sick sinus syndrome and paroxysmal atrial fibrillation (AF) were implanted with Philos DDDR pacemaker. After implantation, auto-Mode-Switch (AMS) function was switched "on" and AF preventive algorithms were "off" in all cases. The number of AMS, atrial premature beats, heart rate and the percentage of atrial and ventricular pacing were recorded by pacemaker diagnostic function for one-month after procedure. AF preventive algorithms function with "middle" (approx 8 bpm) was then switched on and the same parameters as above from the database of pacemaker diagnostic function were collected for additional one month. RESULTS: The symptoms of dizziness, dyspnoea, and palpitation in the majority of patients were dramatically improved regardless of whether the AF preventive algorithms function was switched "on" or "off" after pacemaker implantation. There were no significant clinical changes in most patients when AF preventive algorithms were "on". However, 5 cases (13.2%) had palpitations and short of breath. These symptoms were relieved by changing the algorithms from "middle to slight (approx 4 bpm)". When AF preventive algorithms were switched on, atrial premature beats were reduced significantly (P < 0.05) with a dramatic increase in atrial pacing percentage and heart rate (P < 0.05). However, there was no significant difference in AMS (P > 0.05) between the two groups of AF preventive algorithms function switching "on" and "of", indicating that atrial tachyarrhythmias were not inhibited by anti-atrial fibrillation pacemaker. CONCLUSION: This study suggested that atrial fibrillation and atrial tachycardia were not reduced by implantation of an anti-atrial fibrillation Philos DDDR pacemaker, although atrial premature beats decreased significantly with increasing atrial pacing percentage when AF preventive algorithms were in "middle" and "slight". PMID- 16266460 TI - [The effect of TLR4/NF-kappaB activation and LOX-1 on monocyte adhesion to endothelium]. AB - OBJECTIVE: Recent studies have shown that Toll-like receptor 4 (TLR4), a mediator of for innate immune responses, is involved in the initiation and progression of atherosclerosis. TLR4 activation mediates the expression of chemokines and cytokines through activation of NF-kappaB. We investigated the expression of lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1), intercellular adhesion molecule-1 (CAM-1), E-selectin induced by TLR4/NF-kappaB in human umbilical vein endothelial cells (HUVECs), and their effects on adhesion of monocyte to HUVECs. METHODS: HUVECs were incubated with purified LPS for 24 h. TLR4, LOX-1, ICAM-1, E-selectin mRNA were measured by RT-PCR; the protein expression of TLR4, LOX-1 and activation of NF-kappaB were detected by Western blot; the adhesive percentage between HUVECs and monocytes was determined by direct counting. RESULTS: LPS (1 mg/L) not only enhanced expression of TLR4, activation of NF-kappaB and induction of LOX-1, ICAM-1, E-selectin expression, but also increased the percentage of monocyte adhesion to endothelium. Pretreatment of HUVECs with anti-LOX1, anti-ICAM-1 or anti E-selectin antibodies partly abolished the increase in monocyte adhesion to endothelium. NF-kappaB inhibitor CAPE suppressed LPS-induced these effects. CONCLUSION: TLR4/NF-kappaB plays an important role in monocyte-endothelium adhesion partly through upregulation of LOX-1, ICAM-1 and E-selection expression, which may provide a target for the treatment of atherosclerosis. PMID- 16266461 TI - [Stat 3 signal transduction pathway correlates with proliferation of vascular smooth muscle cells of rats]. AB - OBJECTIVE: To investigate the role of Stat3 (one of the signal transductant and activator) signal transduction pathway in proliferation of vascular smooth muscle cells (VSMCs). METHODS: VSMC was transfected with Stat3 antisense oligonucleotide. ELISA analysis was performed on Stat3 expression in cultured rat thoracic VSMC. Cell proliferation and toxicity assay by Methyl Thiazole (MTT) was used to measure the cell proliferation. The expression of Stat3, p-Stat3, Cyclin D1 and Bcl-x(L) were measured by Western blot. RESULTS: Stat3 protein expression was positive in VSMC. Targeting of Stat3 using antisense oligonucleotide directed against the translation site, resulted in significant growth inhibition and downregulation of Stat3, p-Stat3 and Cyclin D1. CONCLUSIONS: The increase of Stat3 correlates significantly with cell proliferation of VSMC. Cyclin D1 may be the critical target gene in mediating proliferation. Blocking Stat3 signal transduction pathway may inhibit the growth of VSMCs. PMID- 16266462 TI - [Effect of urotensin II on secretion of adrenomedullin from human vascular endothelial cells]. AB - OBJECTIVE: To study the effect of human urotensin II (HU II) on secretion of adrenomedullin (ADM) from human vascular endothelial cells (HVEC) and its mechanism. METHODS: In cultured HVEC, different concentrations of HUII were used to stimulate the ADM secretion from HVEC, and the inhibitors of different signal transduction pathway were used to investigate their effects on ADM secretion. The contents of ADM in medium were determined by radio immunoassay. RESULTS: HUII stimulated secretion of ADM from HVEC in a time-dependent and concentration dependent manner. The contents of ADM in the experiment groups were changed compared with that in control group (P < 0.05). The increase of ADM could be inhibited by inhibitor of extracellular signal-regulated protein kinase (PD(98059)), inhibitor of P38 kinase (SB(202190)), inhibitor of calmodulin (W(7)) and inhibitor of Ca(2+) (nicardipine) (P < 0.05). The inhibition ratio in those groups was 68%, 78%, 24% and 25% respectively. But the inhibitor of Calcineurin (CaN) and inhibitor of protein kinase C (H(7)) had no influence on the secretion of ADM from HVEC (P > 0.05). CONCLUSION: The stimulated effect of HUII on the ADM secretion from HVEC may be mediated by Ca(2+), ERKs, CaM-PK and P38 signal transduction pathways. PMID- 16266463 TI - [Fluid shear stress regulates secretion of tissue-type plasminogen activator in human endothelial progenitor cells]. AB - OBJECTIVE: In order to investigate the role of shear stress in the regulation of endothelial function, we assessed here effects of shear stress on tissue-type plasminogen activator in human endothelial progenitor cells (EPCs). METHODS: The peripheral blood mononuclear cells were separated from healthy adult and inducted into EPCs, which were identified by double staining for the fluorescent labeled acetylated-LDL and lectin. EPCs were seeded on the small diameter artificial vessels, and then divided into four different experimental groups including stationary group, low-flow shear stress group (5 dyn/cm(2)), medium-flow shear stress group (15 dyn/cm(2)) and high-flow shear stress group (25 dyn/cm(2)). The levels of t-PA in EPC culture medium at 0 hour, 5 hours, 10 hours, 15 hours, 20 hours and 25 hours after culture were measured by enzyme-linked immunosorbent assay. RESULTS: The peripheral blood mononuclear cells differentiated into EPCs after induction, which were positively labeled by fluorescent acetylated-LDL and lectin. Shear stress enhanced production of the t-PA by EPCs, which was paralleled to levels and times of shear stress. CONCLUSIONS: Shear stress increases t-PA secretion by human EPCs, suggesting that shear stress not only regulates vascular endothelial function but also participates in the pathogenesis of arteriosclerosis. PMID- 16266464 TI - [The role of isoprenaline-induced, calcium-activated transient outward chloride current in atrial electrical remodeling of rabbit]. AB - OBJECTIVE: To investigate the relationship between the changes of the L-type calcium current (I(Ca, L)) and the calcium-activated transient outward chloride current (I(Cl, Ca)), and the repolarization characteristic of action potential in phase 1 under isoprenaline (ISO) stimulation in atrium myocytes of rabbit. METHODS: Atrium myocytes were obtained by enzymatic dissociation from a section of atrial free wall. The membrane currents and action potential were recorded by the whole-cell patch-clamp technique. RESULTS: After recording I(Ca, L), atrium myocytes were perfused with ISO (1 micromol/L) immediately. Five minutes later, a transient outward current (I(to)) was significantly induced, and the peak of I(to) was gradually increased while I(Ca, L) gradually decreased with increasing in clamp voltage. The I(to) was resistant to 4-AP (3 mmol/L) but sensitive to DIDS (150 micromol/L, Cl(-) channel blocker). This current was blocked by CdCl(2) (200 micromol/L, Ca(2+) channel blocker). The elicited rate of I(to) was 91.67% (P < 0.05). (2) The shape of AP was like an inverse triangle with no plateau in Phase 2 after ISO (1 micromol/L) perfusion. Moreover, compared to the parameters of control group, APD(50) and APD(90) were significantly shortened from (65.4 +/- 4.2) ms and (95.8 +/- 3.8) ms to (12.8 +/- 3.8) ms and (27.0 +/- 4.7) ms, and reduced to 80.46% and 71.87%, respectively (P < 0.01, n = 12). 4-AP (3 mmol/L) had on obvious effect on the shape of AP, however, the plateau of AP in phase 2 was recovered by DIDS (150 micromol/L) perfusion, APD(50) and APD(90) were (41.1 +/- 4.5) ms and (79.6 +/- 3.4) ms respectively. Compared to the parameters of control group, there were no significant differences (P > 0.05, n = 12). These results indicated that ionic transport were changed by ISO perfusion in atrium myocytes and I(to) played an important role in the phase 1 repolarization of AP. CONCLUSIONS: Before ISO administration, we could only observe I(Ca, L) in atrium myocytes of rabbit. After isoproterenol intervention, certain intracellular ionic consistency and membrane ionic channels were changed. Calcium activated chloride channel and I(to2) revealed obvious predominance which shorten APD significantly. Action potential showed a triangle with no plateau, suggesting an electrical remodeling in atrium myocytes. The remodeling of ionic channel is related possibly with the opening of Ca(2+)-activated Cl(-) current, which maybe the electrophysiological base of reentrant atrial tachycardia. PMID- 16266466 TI - [Blood pressure variability and anti hypertensive treatment]. PMID- 16266465 TI - [A cohort study on the relationship between nutrients intake and the incidence of hypertension in middle-aged Chinese]. AB - OBJECTIVE: To explore the relationship between nutrient intake and the incidence of hypertension in middle-aged Chinese. METHODS: A prospective study was conducted in 653 men and women of Beijing and Guanzhou, aged 35-59 years. The subjects had no hypertension or other cardiovascular diseases at baseline risk factor and dietary survey carried out in 1983-1984. The baseline information of nutrient intake was collected by using a 24-hour recall method on three consecutive days. The incidence of hypertension was determined through the follow up survey in 1993-1994. The subjects were categorized into three groups according to tritiles of each baseline nutrient intake and the relative risk for hypertension incidence in each group was calculated by using the logistic regression model with the group of lowest risk as the referent. RESULTS: During the period of 1983-1984 to 1993-1994, 170 among 653 subjects had developed hypertension; 92 were men and 78 were women. After adjustment for other risk factors, the relative risk of hypertension for different protein intake groups was 1.0, 0.87 (95% CI 0.54-1.40) and 0.52 (95% CI 0.32-0.87), respectively, and the p value for trend analysis was 0.011; the relative risk of hypertension for different sodium intake groups was 1.0, 1.12 (95% CI 0.66-1.88) and 1.85 (95% CI 1.09-3.14), respectively, and the p value for trend analysis was 0.015. There was no significant relationship between the other nutrient intake and hypertension incidence. CONCLUSION: Among middle-aged Chinese, protein and sodium may be two of the most important dietary factors affecting the incidence of hypertension. Increasing protein intake and reducing sodium intake might be helpful for the prevention of hypertension. PMID- 16266467 TI - [Complement in atherogenesis]. PMID- 16266468 TI - [The biological treatment of heart failure: current status and prospect]. PMID- 16266469 TI - [A novel LMNA gene mutation E82K associated with familial dilated cardiomyopathy]. AB - OBJECTIVE: To examine the function of the novel mutation E82K in LMNA gene identified in a Chinese family infected by dilated cardiomyopathy. METHODS: (1) One Chinese family infected by dilated cardiomyopathy was chosen for the study. Exons 1-12 of the LMNA gene were screened with both PCR method and the cycle sequencing of the PCR products. (2) cDNA of the E82K mutation or wild type of LMNA gene was transfected into HEK293 cells and the apoptosis of the cells was detected after treatment with 0.8 mmol/L H2O2. RESULTS: (1) A new mutation E82K in LMNA gene was identified in this dilated cardiomyopathy family. (2) Apoptosis was more in the HEK293 cells transfected with E82K mutation than those with empty vector or wild type LMNA gene. CONCLUSIONS: The missense mutation E82K in LMNA gene changed the polar of the amino acid. It showed a malignant phenotype of severe clinical symptoms, early onset, poor survival prognosis and might be associated with atrioventricular conduction block (II degrees-III degrees), suggesting that the E82K mutation in LMNA gene may be a candidate for nosogenesis of dilated cardiomyopathy. PMID- 16266470 TI - [Association of polymorphism in alpha-adducin gene with antihypertensive effect of Hydrochlorothiazide]. AB - OBJECTIVE: To explore the association between G614T single nuclear polymorphism (SNP) of the alpha-adducin gene and the antihypertensive effect of hydrochlorothiazide (HCTZ) in essential hypertensive (EH) patients. METHODS: Eight hundred twenty nine EH patients were given 12.5 mg HCTZ/d for six weeks. Alpha-adducin gene G614T SNP in the tenth exon was determined by PCR-RFLP in 754 patients with complete records. All the patients were grouped according to TT, GT and GG genotypes. RESULTS: After 6 weeks of HCTZ treatment, the decreases in DBP and MAP of patients carrying 614T allele of alpha-adducin were significantly greater than that of those carrying GG homozygotes (P < 0.05). The decreases in SBP and MAP were significantly greater in patients with the TT genotype as compared with GT or GG genotype (P < 0.05). The effective rate of BP fall by HCTZ was higher in patients with TT genotype than those with GT or GG genotype (P < 0.05). Multivariate stepwise regression analysis showed that the TT genotype and the baseline SBP were the two major predictors affecting the decrease in SBP. CONCLUSION: The present study suggests that the alpha-adducin G614T polymorphism is associated with the antihypertensive effect of HCTZ, which is more effective in patients with TT genotype. PMID- 16266472 TI - [The comparison of the effect of enalapril and indapamide on the peripheral blood pressure and central blood pressure through pulse wave analysis]. AB - OBJECTIVES: The purpose of this study was to evaluate the effects of the angiotensin-converting enzyme (ACE) inhibitor enalapril and diuretic indapamide on the peripheral blood pressure and the central blood pressure in Chinese patients with essential hypertension. METHODS: This study was a double blind, randomized study. Informed consent were given by all patients. After 2 weeks of placebo run-in period, 105 patients with mild or moderate essential hypertension were randomized to receive either enalapril (10 mg per day) or indapamide (2.5 mg per day) for 8 weeks. Radial pulse wave recordings were performed in all the patients before the active treatments were given and at the end of the study. Only those patients who have finished 8 weeks of active treatment in both groups were included into the final analysis. RESULTS: One hundred one patients (51 in enalapril group and 50 in indapamide group) completed the study. No significant difference (all P values > 0.05) was found in baseline data between the two groups. After 8 weeks of treatment, all the parameters of pulse wave (except heart rates in both groups and augmentation index in indapamide group) decreased significantly. Comparison of the 2 groups showed that there were no significant differences (all P values > 0.05) in all the parameters of pulse wave except that the central systolic blood pressure, augmentation and augmentation index were significantly lower in enalapril group than in indapamide group. In enalapril group, the reduced values of systolic blood pressure and pulse pressure in central aorta were significantly larger than those in brachial artery. However, the difference was not observed in indapamide group. CONCLUSIONS: Enalapril and indapamide are both similarly effective in reducing peripheral arterial blood pressure. Moreover, enalapril is more effective in reducing central systolic pressure and augmentation index than indapamide. The difference is probably due to the reduction of wave reflection caused by enalapril. PMID- 16266473 TI - [Value of metoprolol injection at dobutamine-atropine stress echocardiography for detection of coronary artery disease]. AB - OBJECTIVE: To investigate the value of metoprolol injection at dobutamine atropine stress echocardiography (DASE) for detection of coronary artery disease (CAD). METHODS: DASE was performed in 72 patients with suspected CAD. All the patients received rapid metoprolol injection immediately after getting peak heart rate at DASE (DASE-Meto) and were subjected to coronary angiography (CAG) within two weeks. Regional wall motion and haemodynamic parameters at peak heart rate during DASE and after metoprolol injection were analyzed, and DASE and DASE-Meto results were compared with CAG. RESULTS: There were 35 patients with CAG positive and 37 negative. The sensitivity, specificity, accuracy and positive and negative predictive values of DASE for detecting CAD were 65.7%, 86.5%, 76.4%, 82.1% and 84.6%, respectively. There were 10 patients with positive result at CAG undetected by DASE but observed regional wall motion abnormality (RWMA) after metoprolol injection. So the sensitivity, specificity, accuracy and positive and negative predictive values of DASE-Meto for detecting CAD were 94.3%, 83.8%, 88.9%, 72.7%, 93.9%, respectively. After metoprolol injection, the symptoms caused by the medicine used in detection were alleviated soon and recovery time was shortened. CONCLUSION: The use of metoprolol at DASE can improve the accuracy and security of CAD detection. PMID- 16266474 TI - [Relationship between coronary arterial remodeling and plaque composition assessed by intravascular ultrasound imaging]. AB - OBJECTIVE: During progression of atherosclerosis, the vessel may develop either positive or negative remodeling. The pathophysiology of vascular remodeling is not fully understood. This study investigated the relationship between plaque characteristics and arterial remodeling using intravascular ultrasound imaging (IVUS). METHODS: A total of 77 patients (male 53, mean age 58 +/- 10 years) who underwent IVUS imaging (ClearView or Galaxy2, Boston Scientific, USA) of culprit vessel were enrolled in this study. Among the 77 patients, 31 presented with stable angina pectoris and 46 presented with acute coronary syndrome. Qualitative assessment of the lesion and quantitative measurement were performed in both stenotic and reference segments. The lesions were classified into soft plaque and hard plaque (including fibrous plaque, calcified plaque and mixed plaque) according to different ultrasound patterns of tissue reflection. The remodeling index (RI) was defined as the ratio of vessel cross sectional area (EEMcsa) of lesion segment to the mean reference EEMcsa. Positive remodeling was defined as RI > 1.0 and negative remodeling as RI < 1.0. RESULTS: Of 77 lesions, 45 (58%) had undergone positive remodeling, and 32 (42%) had negative remodeling. In comparison to the patients with negative remodeling, patients with positive remodeling presented with more acute coronary syndrome (74% vs. 43%, P = 0.006). Both the plaque area and the vessel area were significantly larger in the lesion with positive remodeling than in lesion with negative remodeling. The lesions with positive remodeling were predominantly soft (71% vs. 34%, P = 0.001) and had less calcification [21% vs. 54%, P = 0.003 and (18 +/- 37) degrees vs. (40 +/- 50) degrees, P = 0.027] compared with lesions with negative remodeling. The difference of clinical presentation and plaque characteristics between the patients with different patterns of remodeling is still significant with binary logistic analysis. CONCLUSIONS: Coronary arterial remodeling pattern is related to the clinical manifestation and the composition of plaque. Lesions presented with positive remodeling have a higher prevalence of soft plaque and less calcification. PMID- 16266476 TI - [Plasma concentration of NT-proBNP in patients with acute coronary syndrome correlated with the severity of the diseases?]. AB - OBJECTIVE: To elucidate whether the plasma concentration of NT-proBNP in patients with acute coronary syndrome (ACS) correlated with severity of the diseases and whether NT-proBNP is a reliable biochemical marker correctly indicates the severity of ACS. METHODS: Eighty-nine subjects came from CCU of Cardiology Department of People's Hospital Beijing University from October 2003 to June 2004 and aged 34-85 y (66.89 +/- 11.12 y). In this study the spectrum of ACS only included unstable angina pectoris (UA) and acute myocardial infarction (AMI). Patients with UA were separated into 3 groups by Braunwald classes and those with AMI were separated into 4 groups by Killip classes when their venous blood samples were collected. Plasma concentration of NT-proBNP was measured by enzyme linked immunoabsorbent assay. Data was estimated by SPSS. RESULTS: The concentration of NT-proBNP in patients with ACS was dramatically correlative with the severity of the diseases: with the upgrading of Braunwald classes, the concentration of NT-proBNP in patients with UA increased gradually; in patients with AMI it also raised gradually with the upgrading of killip classes; furthermore, the plasma concentration of NT-proBNP in patients with AIM increased much more than that in patients with UA when they are at the similar NYHA functional class. CONCLUSION: Plasma concentration of NT-proBNP in patients with ACS might be a reliable biochemical marker which can objectively indicate the degree of this diseases. PMID- 16266477 TI - [Clinical research of heart rate turbulence on predictive value in patients with acute myocardial infarction]. AB - OBJECTIVE: To assess the predictive value of heart rate turbulence (HRT) in patients with acute myocardial infarction. METHODS: One hundred and twenty-five patients with acute myocardial infarction were enrolled in this study. During the period from 6 to 21 days after onset of acute myocardial infarction, they were undergone 24-hour Holter recordings to collect the mean RR interval and heart rate variability (HRV) SDNN. The Holter files were processed with software of "HRT! View V0.60-1" to obtain the value of Turbulence Onset (TO) and Turbulence Slope (TS) and the value of "heart rate variability (HRV) SDNN". LVEF and EDD were measured by Ultrasonic Cardiography. Endpoint of follow-up was cardiac death. According to the results, patients were divided into two groups (the "survivors" and the "nonsurvivors"). The predictive value for high-risk patients with acute myocardial infarction was assessed by variables between the two groups. RESULTS: In the period of follow-up (mean 225.4 +/- 99.8 days), 14 patients died and 111 patients survived. In the univariate Cox regression analysis, "TS" was a strong univariate predictor of mortality (hazard ratio 11.46, P < 0.01); "TO" was a relatively weak predictor and the hazard ratio was 2.76 (P > 0.05). Combination of abnormal TO and abnormal TS was the strongest mortality predictor (hazard ratio 26.70, P < 0.01); in the multivariate Cox regression analysis, TS < or = 2.5 ms/RR and EDD > or = 5.6 cm were the independent predictors of mortality with hazard ratios 9.49 (P < 0.01) and 3.64 (P < 0.05), respectively. CONCLUSIONS: The absence of the heart rate turbulence after ventricular premature beats is a very potent post-infarction risk predictor which is independent of and stronger than other known risk predictors. PMID- 16266479 TI - [Clinical study on 29 pacemaker and defibrillator lead fractures]. AB - OBJECTIVE: To analyze major probable causes, locations and remedies of lead fractures in implanted pacemaker and implanted cardioverter defibrillator (ICD) . METHODS: From January 1982 to November 2004, on all the pacemakers and ICDs implanted at this hospital, lead fractures were detected timely by regular telemetric programmed testing, chest radiography and electrocardiography. Appropriate remedial actions were given and follow up visits were done to verify the results. RESULTS: Of all the 4698 pacemakers and 161 ICDs, there were 29 lead fractures on 31 leads with an incidence rate of 0.6%. The incidence rate of lead fracture was 0.9% for pacemakers/defibrillators implanted by subclavian venipuncture, while it was 0.4% for pacemakers/defibrillators implanted by cephalic venotomy (P = 0.04). Fracture locations: 22 cases at the sub clavicle, 6 cases in the pacemaker pocket, and 1 case in the right ventricle. Among the 29 cases, 24 were implanted with additional new leads, 3 cases whose atrial leads were fractured were switched from DDD mode to VVI. All the patients were followed up for an average of 5.6 months +/- 16.1 months. In one patient, the lead was removed through thoracotomy due to infection of exposed lead. CONCLUSIONS: Cephalic venotomy is the first choice for pacemaker and ICD implantation, and the second choice is subclavian venipuncture in the outer part of subclavian vein, which has a higher incidence rate of lead fracture. Most lead fractures are detected at the region of medial subclavian. Regular follow-up can help timely detect fractures, so that the cases can be dealt with properly. PMID- 16266478 TI - [Clinical results of circumferential pulmonary vein linear ablation in 100 patients with atrial fibrillation]. AB - OBJECTIVE: To investigate the feasibility and efficacy of treating atrial fibrillation (AF) with circumferential pulmonary vein (PV) linear ablation guided by 3 dimensional mapping system and single circular mapping catheter. METHODS: From April 2004 to January 2005, PV isolation with circumferential PV linear ablation guided by CARTO system (in 76 patients) or EnSite-NavX system (in 24 patients) was performed in 100 consecutive patients with significantly symptomatic, drug refractory AF. The procedural end-point was complete electrical isolation of bilateral PV. RESULTS: Up to 200 linear circles were produced around each ipsilateral PVs in all 100 cases, and 95.0% (190/200) of PV isolation rate was achieved with a mean procedure time of 150-365 (240 +/- 65) min and a mean fluoroscopy time of 23-61 (37 +/- 12) min, respectively. Eight cases with recurrent AF (8.0%) underwent second session. Cumulative atrial tachyarrhythmias free rate was 85.0% (85/100) during a mean follow-up of 5.5-12 (10.2 +/- 5.7) months. Atrial tachyarrhythmias-free rate was 66.0% (66/100), 82.0% (82/100), 87.0% (87/100), 85.0% (85/100), 85.0% (85/100), and 88.6% (70/79) during the follow up at 1 month, 2 months, 3 months, 4 months, 5 months and 6 months, respectively. There were 2 complications (1 tamponade and 1 PV stenosis), which were rehabilitated after conservative treatment. CONCLUSION: PV isolation with circumferential PV linear ablation guided by 3 dimensional mapping system is safe and effective for treating AF. PMID- 16266481 TI - [Treatment with oral sildenafil in patients with severe pulmonary hypertension after open heart operations]. AB - OBJECTIVE: To evaluate the clinical effects of treatment with oral sildenafil on severe pulmonary hypertension after cardiac surgery. METHODS: From September 2002 to January 2005, oral sildenafil was added to the treatment regime in 27 cases of severe pulmonary hypertension after cardiac surgery. All these cases were given general treatments including intravenous prostaglandin E1 and inhalation of nitric oxide before the use of sildenafil, which did not show obvious effects on decreasing pulmonary pressure. Then a combined treatment [general treatment plus oral sildenafil (1-2 mg/kg, q8h; Pfizer Ltd)] was instituted. Pulmonary artery pressure, systolic pulmonary artery pressure/systolic systemic blood pressure (Pp/Ps) were measured before and every hour after adding sildenafil. RESULTS: One hour after adding sildenafil, the patients' pulmonary artery pressure decreased remarkably (P < 0.01) with no adverse effects on systematic artery pressure. SO(2) and PaO(2) of all cases improved respectively (P < 0.05). One or two days later, the patients' hemodynamics were stable and some patients stopped inhaling nitric oxide and the dosage of prostaglandin E1 decreased. 25 cases stopped use of ventilator and were discharged safely. 2 cases died of multiple organ dysfunction. CONCLUSION: Sildenafil is a highly selective and effective pulmonary hypertension vasodilator, which can be given for the treatment of pulmonary hypertension after cardiac surgery. PMID- 16266483 TI - [Implantation of neonatal cardiomyocytes plus artificial matrix improve heart function in a rat infarct model]. AB - OBJECTIVE: To investigate whether injectable engineering heart tissue (EHT) can survive and improve heart function after transplantation into infarct area. METHODS: Ventricular cardiomyocytes from 1-3 day-old Sprague-Dawley (SD) rats were isolated by using trypsin method, and then labeled and cultured. The left coronary of female SD rats was ligated to create a myocardial infarct model. Three weeks later, the qualified animals were randomized into four groups: EHT group (n = 12), which were transplanted with both cardiomyocytes and matrix; cell transplantation group (n = 12); matrix group (n = 12), control (n = 11). Four weeks after implantation, echocardiography and Langendorff model were used to assess heart function, and then the hearts were harvested for pathological examination. Meanwhile polymerase chain reaction (PCR) was performed to detect SRY gene on Y chromosome. RESULTS: The grafted cells were identified in both EHT and cell transplantation group by either pathology or PCR. But in EHT group, transplanted cells formed more condensed tissue, and produced definite connected protein. Data of fraction shortness from echocardiography are showed as follows: EHT group, (22.82 +/- 3.44)%; cell transplantation group, (20.55 +/- 4.11)%, matrix group, (17.05 +/- 4.57)%; control, (19.80 +/- 3.98)% (P = 0.012). Langendorff examination revealed significant differences among four groups when left ventricular balloon volume was at the level of 0.06 ml, 0.08 ml and 0.10 ml, in which EHT group had the highest developed pressure and dp/dt. CONCLUSION: It is feasible to fabricate injectable EHT in vitro. The fabricated EHT could survive in the myocardial infarct area after transplantation in a rat model and improve heart function due to better histological configuration. PMID- 16266486 TI - [Relationship between expression of Smad and ventricular remodeling after myocardial infarction in rats]. AB - OBJECTIVE: To investigate the relationship between expression of Smad3, Smad7 and ventricular remodeling in rats after myocardial infarction. METHODS: Myocardial infarction was induced by left anterior descending coronary artery ligation in rats (n = 11) and sham-operated rats were used as control (n = 10). The rats were sacrificed 8 weeks later. Heart weight/body weight (HW/BW), mean blood pressure, left ventricular end diastolic pressure (LVEDP), collagen content in the un infarcted area were examined. The mRNA levels of transforming growth factor (TGF)beta(1), Smad 3, Smad7 were determined by RT-PCR. RESULT: Compared with controls, the level of HW/BW, LVEDP and collagen content were significant increased. The mRNA expression of TGFbeta(1) and Smad3 was significantly increased in areas of myocardial infarction, border of the infarction, interventricular septum and right ventricle. The expression of Smad7 mRNA in these areas was decreased. CONCLUSION: These results indicated that TGFbeta(1) Smads signaling was correlated to the ventricular remodeling after myocardial infarction. Smad3 might promote the process while Smad7 inhibit the process. PMID- 16266484 TI - [The mechanism of stem cell factor in bone marrow stem cell transplantation to heart]. AB - OBJECTIVE: To study the mechanism of stem cell factor (SCF) in bone marrow stem cells heart transplantation (BMT) and the influence of bone marrow mobilization on the transplantation efficacy. METHODS: Rats with acute myocardial infarction (AMI) accepted BMT. The SCF expression in the bone marrow was measured by RT-PCR after the operation. Then bone marrow stem cells with different SCF levels for the transplantation were used and the cardiac function was compared by using echocardiography. The SCF protein expression in the heart, plasma and bone marrow was detected by ELISA. RESULTS: SCF expression level decreased significantly 1 week after AMI (P < 0.01), but it didn't decrease in those accepting BMT. Though the rats accepted BMT with bone marrow stem cells from different sources, the cardiac function showed no difference (P > 0.05). After BMT, the SCF protein level in the plasma decreased significantly (P < 0.05). CONCLUSIONS: BMT may make mobilization through SCF. Bone marrow stem cells from rats with AMI and also those with myocardial infarction plus BMT therapy can also be used for the transplantation into heart, and have no influence on cardiac function improvement. PMID- 16266487 TI - [The ultrastructure change of cardiomyocyte in Athlete's Heart]. AB - OBJECTIVE: This study is to see the pathologic change of cardiac myocyte in Athlete's Heart, and explore the mechanism of the pathologic change. METHODS: Fifteen male SD rats were separated randomly into control group (without any exercise), aerobic exercise group (Ae group, swimming for 75 min every day), and overloading exercise group (Oe group, swimming for 180 min with a loading of 5 percent of body weight every day). After 5 days per week for 12 weeks, swimming stopped, the rat hearts were prepared to specimens and examined under Transmission Electron Microscope. RESULTS: The Ae group, the number and volume of mitochondria increased, and the membrane of mitochondria remained entire. Few of dense bodies were found in cytoplasm. The nucleus envelopes of expansion nucleus appear as dentition. These changes were considered as the adaptation to exercises. At the same time, some pathologic changes of the cardiac myocytes similar to senescence also appeared, such as mitochondria expanse, the crista disorder or disappearance, unclear mitochondria membrane, many dense bodies in cytoplasm, nucleus disfiguration and chromatin collection at edge. CONCLUSION: After exercise training, some pathologic changes of cardiac myocyte also occur with physiological changes. With the raise of exercise intension, the pathologic changes become more obvious, even appearance of cardiac myocyte death. PMID- 16266488 TI - [Role of peroxisome proliferators activated receptor gamma in mediating proliferation of rat vascular smooth muscle cells induced by advanced glycation end products]. AB - OBJECTIVE: To observe the effect of pioglitazone on advanced glycation end products (AGEs)-induced proliferation of vascular smooth muscle cells (VSMCs) and expression of peroxisome proliferators activated receptor gamma (PPARgamma). To investigate the possible role of PPARgamma in mediating AGEs induced proliferation of VSMCs. METHODS: Primary cultures of smooth muscle cells from rat aorta were exposed to AGEs of different concentrations and different times prior to co-treatment with pioglitazone and AGEs. 3-(4, 5-dimethylthiazol-2-yl)-2, 5 diphenyl tetrazolium bromide (MTT) assay was adopted for the quantification of the cell proliferation ratio and PPARgamma expression was determined by RT-PCR and Western immunoblotting. RESULTS: AGEs increased the proliferation of VSMCs. AGEs treatment to VSMCs decreased mRNA and protein levels of PPARgamma in a time- and dose-related manner (P < 0.05). Pioglitazone inhibited the AGEs-induced proliferation of VSMCs in vitro. CONCLUSIONS: Activating PPARgamma in VSMCs, pioglitazone may play a role in anti atherosclerosis. The reduction in PPARgamma expression may be implicated in vascular smooth muscle cells proliferation and pathogenesis of atherosclerosis in patients with diabetes mellitus. PMID- 16266490 TI - [An nonselective cation current in rabbit ventricle myocytes]. AB - OBJECTIVE: Currents contributing repolarization in rabbit ventricular myocyte are very complex since the I(To.s) covers almost the whole repolarization phase of the action potential. The other components of repolarizing currents, as I(Kr) and I(Ks) are small. The purpose of this study is to investigate whether or not there are other currents in rabbit ventricular repolarization. METHODS: Ion currents of rabbit ventricular myocyte were recorded using the whole-cell patch-clamp technique. RESULTS: In the present work, an nonselective cation current was identified by replacing the K(+) with Cs(+) in the bathing and pipette solutions. The outward current elicited by depolarizing potentials could be inhibited by Gd(3+), an effective inhibitor of nonselective cation currents. Depleting Ca(2+) and Mg(2+) in the bathing solution, the amplitudes of this outward current increased by 40%-116% at +60 mV, and adding 2 micromol/L insulin to the solution (with normal concentration of Ca(2+) and Mg(2+) in Tyrode's solution), the amplitude increased by 30%-60% at +60 mV. CONCLUSION: It is suggested that a nonselective cation current in rabbit ventricular myocytes may play an important role in the repolarization of the action potential in rabbit ventricle. Changes of nonselective cation current will lead to induce or inhibit arrhythmia. PMID- 16266498 TI - [Application of optical coherence tomography in the detection of atherosclerosis]. PMID- 16266491 TI - [A cross sectional survey on serum lipid level and its influencing factors in children aged 3-14 years in Guangdong province]. AB - OBJECTIVE: To investigate the serum levels of cholesterol, triglyceride and high density lipoprotein cholesterol in children aged 3-14 years and its influencing factors. METHODS: The cross-sectional survey study population was a representative sample from Guangdong province obtained by multi-stage randomized cluster sampling. Serum lipids in 6188 children aged > or = 3 years were assayed using automatic biochemical instrument. The data of social and demographic status were collected by face-to-face interview, and height and weight were obtained by physical examination. RESULTS: The age-standardized and region-weighted means of serum total cholesterol (TC), triglyceride (TG), and high density lipoprotein cholesterol (HDL-C) were 0.80 mmol/L, 3.50 mmol/L and 1.28 mmol/L, respectively. For the mean of TG, there was no difference between metropolitan and middle city, nor between rich county and poor county. For TC, it was the highest in metropolitan, and there was no difference between rich and poor county. For HDL C, the difference existed between every two regions. The age-standardized and region-weighted prevalence of high TG, high TC and low HDL-C were 2.2%, 2.1% and 8.0%, respectively. Metropolitan, rich county, low weight and age between 7.0-9.9 years are protecting factors for high TG, and the number of family between 3-4, age between 7.0-9.9, metropolitan, middle city and poor county are risk factors for high TC. Male, family income per year between 800-9999 RMB, middle city, rich county are protecting factors for low HDL-C. CONCLUSION: The prevalence of abnormal serum lipid was still low compared with other regions in China. The region, number of family member, age and sex may be the important factors influencing on serum lipid levels. PMID- 16266499 TI - [Coronary slow flow phenomenon]. PMID- 16266502 TI - [Predictive equations and analysis of influence factors of lung ventilation based on a large occupational population in North China]. AB - OBJECTIVE: To establish suitable predictive equations of lung function for occupational population in North China. METHODS: A total of 5 002 on the job or retired healthy adults from five enterprises in North China with category of mild or moderate work intensity underwent spirometry using a Chest HI-198 spirometer and the procedures recommended by the American Thoracic Society, were a sample. RESULTS: The data of 3 913 subjects were used. A normal distribution of our data was shown using the normality test and distribution curve. Univariate analysis showed that both age and height were significantly correlated with FVC, FEV(1), FEV(1)/FVC (%) and MMF. Further multiple linear stepwise regression analysis indicated that the levels of FVC, FEV(1), FEV(1)/FVC (%) and MMF were highly influenced by age, height, and weight rather than chest circumference. Thus, only age, height, and weight were introduced into our regression equations. Data from the studied subjects and other source were utilised to examine the validity of the equations and a high accordance rate (> 90%) was obtained. No significant difference (P > 0.05) was found in the predictive values between the simplified equations and equations in which more variables were included. CONCLUSION: The studied predictive equations for male non-smokers, female non-smokers, and male smokers were established based on data from a large occupational population. These equations should be more applicable for evaluating lung ventilatory function of occupational populations in North China. PMID- 16266503 TI - [Application of c-- reactive protein in diagnosis and treatment of nosocomial pneumonia with coal workers' pneumoconiosis]. PMID- 16266504 TI - [Experimental study on the DNA damage of NIH/3T3 cells induced by nickel-refining dusts]. AB - OBJECTIVE: To study the effects of nickel-refining dusts on DNA damage in mouse NIH/3T3 cell. METHOD: DNA damage of mouse NIH/3T3 cell induced by two nickel refining dust samples was determined with single cell gel electrophoresis (SCGE) technique. RESULTS: (1) Under the condition of the same treatment time, the tailed cell (%) of NIH/3T3 cells increased with the increase in doses of nickel refining dusts (35.5%, 69.7%, 85.2%, 41.3%, 75.7%, 89.2% respectively except for sample 2, 50 microg/ml, 24 h group), and DNA strand breaks reached the peak value at 4 h of exposure; (2) When we treated the NIH/3T3 cells with the same dose of nickel-refining dusts, the tail rate at 4 h was higher than those at 2 h and 24 h of exposure; (3) Both sample 1 and sample 2 with different doses of nickel refining dusts could induce higher comet tail, DNA%, tail length (except for 12.5 microg/ml), extent of TM (except for sample 1, 12.5 microg/ml) than in control group (P < 0.05). The DNA damage range was significantly increased in different tested doses of nickel-refining dusts and the damage range reached the peak value when the cells were treated for 4 h. CONCLUSION: Nickel-refining dusts could induce different degree DNA damage in NIH/3T3 cells. PMID- 16266506 TI - [Follow-up study on psychic and neuropathic delayed effects of acute organophosphorus pesticides poisoning]. AB - OBJECTIVE: To study the delayed effect on neuropsychopathy and its related factors after acute organophosphorus pesticides poisoning (AOPP). METHODS: Two hundred and fifty-seven cases of AOPP in the observation period were chosen to follow-up 2 months later from the 4 county hospitals in Shandong Province where the incidence of organophosphorus pesticide poisoning is high. RESULTS: Nine cases of organophosphate induced delayed polyneuropathy (OPIDP) were found and the incidence rate was 3.5%. The occurrence of OPIDP were related to the need for emergent artificial respiration, and the degree of poisoning, and the kinds of organophosphorus pesticides (Ops). The positive rate of symptoms of peripheral nerves, central nerves and psychogeny except auditory and visual hallucination after poisoning was significantly higher than that before (P < 0.05). The patient's situation of health, economy and work became statistically worse (P < 0.05). CONCLUSION: We found some had delayed effects on neuropsychopathy after AOPP which could debase the patient's life quality. The control measure should be administered as early as possible. PMID- 16266505 TI - [Inhibition of the pathway of benzo (a) pyrene-induced cell cycle changes by all trans retinoic acid in lung fibroblast]. AB - OBJECTIVE: To investigate the reverse effect of all-trans retinoic acid (ATRA) on Benzo (a) pyrene (B (a) P)-induced cyclin D1, CDK4, E2F-1 and E2F-4 expression and cell cycle progression in human embryo lung fibroblast (HELF). METHODS: After HELF cells was treated with ATRA, they were exposed to 2 micromol/L of B (a) P. Western blotting was employed to detect protein expression level; the RNA transfection techniques was used to investigate ATRA-induced signal pathway; flow cytometry was used to detect cell cycle progression. RESULT: After treatment with 2 micromol/L B (a) P for 24 h, the expression of cyclin D1 and E2F-1 were both increased significantly in HELF; the expression of E2F-4 and CDK4 were not changed markedly; pretreatment with 0.1 micromol/L ATRA for 24 h could efficiently decrease B (a) P-induced overexpression of cyclin D1 and E2F-1; stimulation to antisense cyclin D1 or antisense CDK4 by B (a) P could significantly impair E2F-1 up-regulation; pretreatment with ATRA, cells with antisense cyclin D1 or antisense CDK4 showed a less decrease in B (a) P-induced overexpression of E2F-1 compared to similarly treated control cells; flow cytometry analysis showed B (a) P promoted cell cycle progression from G(1) phase to S phase, while pretreatment with ATRA could inhibit B (a) P-induced cell cycle progression by an accumulation of cells in the G(1) phase. CONCLUSION: ATRA could block B (a) P-induced cell cycle promotion through cyclin D1/E2F-1 pathway in HELF. PMID- 16266507 TI - [The reporting system of acute pesticides poisoning and general situation of pesticides poisoning in China]. AB - OBJECTIVE: To introduce the reporting system of acute pesticides poisoning and analyze epidemiologic characteristics of pesticides poisoning from reported cases in China. METHODS: Case reports in the data base of reporting system for occupational diseases were computed by Excel for windows and statistical significance by SAS 6.12. RESULTS: A total of 108 372 cases were reported from 1997 to 2003. Among them, the incidence of occupational poisoning, and non occupational poisoning accounted for 25.39%, and 74.61% respectively. The fatality rate was 6.86%. The average age was 36.83 years for all pesticides poisoning patients, and 15-59 years old patients accounted for 84.11%. Among 0-14 years old non-occupational poisoning patients, 0-4 years children accounted for 33.51%. Male patients were in the majority in occupational pesticides poisoning, female in non-occupational. Insecticides especially organophosphorus insecticides such as methamidophos, parathion, and omethoate comprised a higher proportion, accounting for 86.02% of the pesticides poisoning. CONCLUSION: More attention should be paid to pesticides poisoning by the government and medical workers engaged in public health. PMID- 16266508 TI - [A clinicopathological study of 16 autopsy cases of anthracosilicosis with lung cancer]. AB - OBJECTIVE: To investigate the clinicopathological characteristics of anthracosilicosis complicated with lung cancer. METHODS: Tissue specimens from 16 autopsy cases of 0(+) anthracosilicosis complicated with lung cancer were retrospectively studied by hematoxylin-eosin, histochemical, and immunohistochemical staining. RESULTS: All of 16 patients were male. The patient's age ranged from 46 to 57 years (average: 52.8 years). The dust-exposure time were over 25 years. The pneumoconiosis and dust fibrosis of different degrees in the lung were found. The positive detected rate of coal silicotic nodules was 93.75% (15/16). Among 16 cases of lung cancer, there were 5 cases of squamous cell carcinoma, and 5 cases of small cell undifferentiated carcinoma, 3 cases of bronchioloalveolar carcinoma, 2 cases of adenocarcinoma and 1 case of adenosquamous carcinoma. The typical pathological changes of anthracosilicosis complicated with lung cancer were: the cancer tissue was located at the side of coal dust fibrous focus and fibrosis lesion, or mixte with silicotic lesion. CK, EMA and CEA were positively expressed in most of the tumor cells, while vimentin was positive in the fibrocyte of dust fibrosis. CONCLUSIONS: The occurrence of some lung cancer may be related with fibrosis. The dust-exposed workers can suffer from lung cancer which is histologically identical to the general lung tumor. PCNA and Ki67 may be a prognostic index for anthracosilicosis with lung cancer, while vimentin may be a marker for the examination of dust fibrosis in anthracosilicosis. PMID- 16266510 TI - [One case of tetramine poisoning due to occupational exposure]. PMID- 16266509 TI - [Analysis of serum glutathione S-transferase and urinary 8-hydroxy-2 deoxyguanosine in coke oven workers]. AB - OBJECTIVE: To investigate the application of serum glutathione S-transferase (GST) and urinary 8-hydroxy-2-deoxyguanosine (8-OHdG) as the monitoring biomarkers for coke oven workers exposed to polycyclic aromatic hydrocarbons (PAHs). METHODS: 47 male coke oven workers and 31 male control workers were investigated. Urinary 8-OHdG and serum GST were analyzed using high performance liquid chromatography (HPLC) with electrochemical detection and test kit. Urinary 1-hydroxypyrene (1-OHP) as internal exposure of PAHs was also determined simultaneously by alkaline hydrolysis and HPLC. RESULTS: The values of urinary 1 OHP, serum GST and urinary 8-OHdG were reported as median with interquartile range (P(25)-P(75)). Urinary 1-OHP [5.7 (1.4-12.0) micromol/mol Cr], serum GST [22.1 (14.9-31.2) U/ml], and urinary 8-OHdG [1.9 (1.4-15.4) micromol/mol Cr] in coke oven workers were significantly higher than in control workers [3.0 (0.5 6.4) micromol/mol Cr (P < 0.05), 13.1 (9.5-16.7) U/ml (P < 0.01), and 1.3 (1.0 4.0) micromol/mol Cr (P < 0.05) respectively]. Categorizing by smoking status, significant differences in urinary 1-OHP and serum GST were found only in smokers among coke oven workers compared to control workers (P < 0.01), and 8-OHdG levels only in non-smokers (P < 0.01). Additionally, there was significant correlation between urinary 1-OHP and serum GST activity (r(s) = 0.31, P < 0.01, n = 78). The multiple logistic regression analysis showed that coke oven workers were at the higher risk of having GST activities above 16.7 U/ml (OR = 13.2) and 8-OHdG levels above 1.8 micromol/mol creatinine (OR = 4.4). High body mass index was an independent factor to affect urinary 8-OHdG levels. CONCLUSIONS: The elevated serum GST activities and increased oxidative DNA damage were found in the coke oven workers. Occupational exposure and smoking interact on each other. Serum GST may be used as a biomarker for assessing the exposure of PAHs. Assay of urinary 8 OHdG may be useful for evaluating the risk of lung cancer in coke oven workers. PMID- 16266511 TI - [Study of trichloroethylene-induced apoptosis in normal human epidermis keratinocytes in vitro]. AB - OBJECTIVE: To investigate the apoptosis-inducing effect of trichloroethylene (TCE) on cultured normal human epidermis keratinocytes (NHEK) in vitro. METHODS: NR(50) values (the concentration of neutral red absorbed is reduced to 50%) of TCE on NHEK were assayed by neutral red uptake (NRU), and the administered dose of TCE was determined. Lipid peroxidation (LPO) and oxidative stress were assessed by measurement of malondialdehyde (MDA) contens and superoxide dismutase (SOD) activity. Transmission electron microscope (TEM) were used to observe morphologic changes, flow cytometer (FCM) was used to measure DNA contents and calculate cell apoptosis rate and proliferation index (PI). RESULTS: NR(50) values of TCE on NHEK was found to be 4.53 mmol/L (95% CI: 3.92-5.13 mmol/L). The increase in MDA content and inhibition of SOD activity in a concentration dependent manner were shown after NHEK was treated with a series of dose of TCE 4 h later, and typical morphologic changes of apoptosis were also observed by TEM examination. FCM analysis revealed a sub-G(1) peak in the apoptotic cells. The apoptotic rate in TCE 0.125, 0.500, 2.000 mmol/L exposed groups (31.83%, 38.63%, 44.35%, respectively) were significantly higher than that in blank control (18.42%), while PI in TCE 0.125, 0.500, 2.000 mmol/L group (3.26%, 2.48%, 2.07%, respectively) were significantly lower than that in blank control (4.99%). CONCLUSION: TCE may induce apoptosis of cultured NHEK in vitro, and inhibit cell proliferation through lipid peroxidation and oxidative stress. PMID- 16266512 TI - [Electrophysiological study of 16 patients with severe N-hexane neuropathy]. AB - OBJECTIVE: To observe electrophysiological changes of severe N-hexane neuropathy getting active therapies and discuss its prognosis. METHODS: A follow-up study involved 16 adult severe N-hexane neuropathy patients who got active therapies was performed. EMG in right muscle of thenar, tibial muscle, and vastus medialis, NCV in right median nerve, common peroneal nerve, and sural nerve were determined and analyzed before treatment and in the first, the third, the ninth, and the twenty-fourth month after treatment. RESULTS: The electrophysiology in severe N hexane neuropathy patients showed that the voluntary potential during muscle relaxation increased by 25.0%; the motor unit potential time limit prolonged by 20.8%, and the amplitude increased by 12.5%, and multiphasic wave increased by 16.5% during mild contraction; the raise decreased by 25.0% during strong contraction. In control group, the MCV, SCV, SNAP, DML, and CMAP of median nerve were (54.63 +/- 5.33) m/s, (59.25 +/- 6.45) m/s, (26.53 +/- 6.32) microV, (3.96 +/- 0.65)ms, and (9.89 +/- 2.30) mV respectively, the MCV, CMAP, DML of common peroneal nerve were (48.49 +/- 3.25) m/s, (5.47 +/- 1.77) mV, (5.20 +/- 1.27) ms respectively, and the SCV, SNAP of sural nerve were (63.21 +/- 9.30) m/s, (4.63 +/- 1.29) microV respectively. Severe N-hexane neuropathy patients presented significantly different abnormalities in the NCV and EMG (P < 0.01). The MCV, SCV, SNAP, DML, CMAP of median nerve were (46.00 +/- 4.32) m/s, (40.66 +/- 2.65) m/s, (7.98 +/- 1.05) microV, (4.28 +/- 0.63) ms, and (6.32 +/- 1.54) mV respectively. The MCV, CMAP, DML of common peroneal nerve were (48.49 +/- 3.25) m/s, (3.21 +/- 1.99) mV, (7.32 +/- 1.65) ms respectively. The SCV, SNAP of sural nerve were (36.48 +/- 5.20) m/s, (2.15 +/- 1.22) microV respectively. These parameters gradually recovered to normal levels in 24 months after treatment. CONCLUSION: The electrophysiological abnormalities in severe N-hexane neuropathy patients can restore after treatment, and clinical prognosis is good. PMID- 16266514 TI - [Effects of silica on the expression of plasminogen activator inhibitor-1 and activator protein-1 in human alveolar epithelial cells type II]. AB - OBJECTIVE: To investigate the mechanism of silicosis by observing the effects of silica on the expression of plasminogen activator inhibitor-1 (PAI-1) and activator protein-1 (AP-1) in human alveolar epithelial cells type II (A549). METHODS: A549 cell and SiO(2) (200 microg /ml) were co-cultured for 0, 4, 8, 16 and 24 h respectively. The reverse transcriptase polymerase chain reaction (RT PCR), Western blotting and SP immunocytochemistry were used for detections of the PAI-1 mRNA and protein expression. The nucleoprotein and total protein expression of AP-1 were investigated by Western blotting. RESULTS: The expression levels of PAI-1 mRNA and protein were increased in a time-dependent manner(r(mRNA) = 0.911, r(protein) = 0.902, P < 0.05). The expressions of PAI-1 mRNA and protein in experimental groups were higher than that in control group (P < 0.05) and was the highest in 24 h group [(0.73 +/- 0.01) vs (0.36 +/- .03)]. The nucleoprotein expressions of c-jun/c-fos in experimental groups were also higher than in control group (P < 0.05), and the nucleoprotein expression level of c-jun was the highest in 4 h group [(1.54 +/- 0.02) vs (0.56 +/- 0.03)]; the nucleoprotein expression level of c-fos was the highest in 8 h group [(0.36 +/- 0.01) vs (0.15 +/- 0.01)]. Both c-jun and c-fos expression were decreased after 16 h, but the total protein expression of c-jun/c-fos had no difference in all experimental groups. The positive signal of PAI-1 was located in cytoplasm and nucleus. CONCLUSION: SiO(2) could induce PAI-1 expression of A549 in a time-dependent manner, and AP-1 activation can be observed in early time. PMID- 16266515 TI - [Abnormalities of microsatellite in transitional cell carcinoma of urinary bladder related with aromatic amine exposure]. AB - OBJECTIVE: To study the microsatellite abnormalities of the aromatic amine exposure-associated transitional cell carcinoma (TCC) and sporadic TCC of urinary bladder, and to evaluate the potential of microsatellite analysis on detection of this diseases. METHODS: Based on our previous investigations, 5 microsatellite markers (D17S695, D9S162, D3S1295, DBH and D3S1234) that had high frequencies of loss of heterozygosity (LOH) in sporadic TCC, were selected for analysis with the bladder lesions derived from 16 patients with aromatic amine exposure history. The microsatellite analysis with urine sediments from the post-operated patients was also carried out. RESULTS: There was at least one informative marker out of the 5 microsatellite foci showed polymorphism in the DNA derived from 16 patients examined. Within 87.50% (14/16) patients, LOH was detected in the bladder lesions at least with one microsatellite marker. The LOH frequency of D3S1295 was higher in occupational TCC patients than that in sporadic TCC patients. The diagnostic accordance rate of patients showed LOH in at least one microsatellite marker with patients diagnosed by pathology was 81.25% (13/16). In the urine sediments from 8 TCC post-operated patients, LOH was found at least with one microsatellite marker. CONCLUSION: There could be a different LOH pattern in aromatic amine exposure-associated TCC, and genes near D3S1295 might play a role in the occupational exposure-associated TCC. PMID- 16266516 TI - [DNA damage induced by methyl tertiary-butyl ether in vivo and in vitro]. AB - OBJECTIVE: To investigate the mechanism of methyl tertiary-butyl ether (MTBE) induced animal carcinoma. METHODS: Single cell gel electrophoresis assay (SCGE), DNA cross-links test and unscheduled DNA synthesis (UDS) assay were conducted with cultured rat type II pneumocytes and rat hepatocytes in vitro. Except UDS assay, the same experiment was performed in hepatocytes, renal cells and pneumocytes of mice administrated MTBE by inhalation at 0, 108, 1,440 and 4,968 mg/m(3) for 20 consecutive days. Simultaneously, the contents of malondialdehyde (MDA) in homogenates of lung and kidney were determined. RESULTS: The lengths of DNA migration in mice hepatocytes at 1,440, 4,968 mg/m(3) of MTBE, renal cells at all doses of MTBE, and pneumocytes at 4,968 mg/m(3) were greater than those in negative controls. There was dose-effect relationship between the concentration of MTBE and hepatocytes DNA migration lengths in mice (r = 0.997, P = 0.003). MTBE of 1,440 and 4,968 mg/m(3) contributed to a rise in MDA of renal homogenates in female mice (P < 0.05). MTBE above 0.050 mmol/L caused greater DNA migration in cultured rat type II pneumocytes and rat hepatocytes in vitro (P < 0.05), and also with dose-effect relationship (r(lung) = 0.967, r(liver) = 0.963, P < 0.05)). In UDS assay, DNA synthesis of rat type II pneumocytes and rat hepatocytes were increased at the concentration of 5.0 mmol/L and 10.0 mmol/L of MTBE. CONCLUSION: MTBE has some genotoxicity on DNA, and the single strand breaks of cell and lipid peroxidation may be one of the possible mechanism of MTBE induced hepatic and renal tumors of animal. PMID- 16266517 TI - [An accidents of acute hydrogen sulfide poisoning]. PMID- 16266518 TI - [Biological responses of tin mine particles and their association with adverse effects on health in tin mine]. AB - OBJECTIVE: To evaluate the biological and toxicity of tin mine particles mixed with crystalline silica using an in vitro test, and to compare to the pathogenesis of pneumoconiosis and lung cancer. METHODS: Respirable particle samples were sampled from four tin mines, in which elevated mortality of pneumoconiosis and lung cancer were reported in miners exposed to particles. Alveolar macrophages (AM) are considered as the target cells of primary dust effects. The samples were then measured in 15, 30, 60 and 120 microg particle per 106 AM for cytoxicity with the release of glucuronidase, lactate dehydrogenase, for reactive oxygen damage with H2O2 release, and for ability to induce fibrosis using the secretion of tumor necrosis factor-alpha (TNF-(alpha) in guinea pig and/or rat am. pure quartz (dq12) and corundum were used as controls. RESULTS: The results showed the samples from tin mines caused a higher cytoxicity when compared to corundum, yet lower when compared to quartz. However, reactive oxygen species release induced by the samples were significantly higher than that induced by quartz and corundum. Beside particle samples induced higher TNF-alpha secretion than corundum, samples from Limu tin mine also induced greatly higher TNF-alpha levels than that induced by pure quartz, even in the lowest concentration. The results from epidemiological research show that high incidence of silicosis among tin miners. And standardize mortality from all cancer (SMR = 1.58, 95% CI: 1.39-1.76) and lung cancer (SMR = 3.17, 95% CI: 2.59-3.76) are higher than national average level. CONCLUSION: The results from in vitro test may reasonable interpret high risk of pneumoconiosis and lung cancer in tin miners. The in vitro multidimensional reaction patterns of AM can be used to screen workplace particles for adverse effects to health. PMID- 16266519 TI - [Effect of the pre-hospital systematic treatment on prognosis patients of with severe acute organophosphorus pesticide poisoning]. AB - OBJECTIVE: To investigate if the duration from poisoning to treatment (no treatment period) is related to the prognosis of patients with severe acute organophosphorus pesticide poisoning (SAOPP). METHODS: One hundred and seventy four patients with the pre-hospital systematic treatment served as the treatment group while 160 patients going to the hospital by themselves without treatment or rejecting gastrolavage served as the control group. Patients in both groups were treated by gastrolavage, pralidoxime chloride, atropine and other expectant treatment. The duration of no treatment period, death, and severe complication were observed. The time of disappearance of symptoms, the recovery time of acetyl cholinesterase (AChE), atropinization time, atropine dosage, pralidoxime chloride dosage, naloxone dosage, hospitalization days and other targets were also observed. RESULTS: The duration of no treatment period in treatment group [(1.2 +/- 0.3) h] was significantly shorter than that in control group [(2.8 +/- 0.5) h, (P < 0.01)]. The mortality rate in treatment group was 6.32% while that in control group 22.5% (P < 0.01). The incidence of respiratory failure, heart injury, brain injury, atropine poisoning, intermediate syndrome, liver injury in treatment group (12.64%, 5.75%, 8.62%, 1.72%, 4.60%, 5.17% respectively) were lower than those in control group (25.63%, 13.75%, 17.50%, 6.25%, 7.50%, 9.38% respectively, P < 0.05 or P < 0.01). The time of symptoms disappearance, the recovery time of AChE, atropinization time, atropine dosage, pralidoxime chloride dosage, naloxone dosage, hospitalization days in treatment group were significantly superior to those in control group (P < 0.05 or P < 0.01). CONCLUSION: The pre-hospital systematic treatment can improve the prognosis of the patients with SAOPP, which is worth popularizing and using. PMID- 16266520 TI - [Change of urinary microalbumin in patients with acute organophosphorus pesticide poisoning]. PMID- 16266522 TI - [Investigation on an accident of death induced by bromoxynil poisoning]. PMID- 16266523 TI - [Relationship between cerebral infarction and nosocomial pneumonia with coal workers' pneumoconiosis]. PMID- 16266524 TI - [Investigation on an accident of trichloroethylene-induced medicamentosa like dermatitis]. PMID- 16266525 TI - [The effect of pyraloxime methylchloride to myocardial tissue induced by methamidophos poisoning in rats]. PMID- 16266528 TI - [Study on clinical treatment of acute bromoxynil poisoning]. PMID- 16266529 TI - [Eight cases of acute phosphine poisoning]. PMID- 16266530 TI - [One case of extrinsic allergic alveolitis with librarian]. PMID- 16266532 TI - [Review of leukemia induced by benzene poisoning]. PMID- 16266533 TI - [The use of induced sputum in treatment of occupational asthma]. PMID- 16266535 TI - [Improving the nutrition and health status of Chinese people, for facilitating the development of social economy]. PMID- 16266536 TI - [Prevention of vaccine derived poliovirus transmission]. PMID- 16266537 TI - [The plasma lipids level in adults among different areas in China]. AB - OBJECTIVE: To study plasma total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C) levels of Chinese adults among different areas in China. METHODS: Analysis of fasting plasma lipid in 49,252 subjects aged 18 years and above was made. RESULTS: The mean TC, TG and HDL-C were 3.81 mmol/L, 1.10 mmol/L and 1.30 mmol/L, respectively. The mean TC, TG and HDL-C is 3.96 mmol/L, 1.16 mmol/L, 1.30 mmol/L in adults in urban and 3.75 mmol/L, 1.07 mmol/L and 1.30 mmol/L in adults in rural areas. In age groups of 18 - 44 years, 45 - 59 years and 60 years above, the mean values were 3.70 mmol/L, 4.09 mmol/L, 4.21 mmol/L for TC; 1.07 mmol/L, 1.21 mmol/L, 1.20 mmol/L for TG; and 1.29 mmol/L, 1.33 mmol/L, 1.33 mmol/L for HDL-C. CONCLUSION: The results of this study might provide the national representative data of plasma lipid level. TC and TG levels of adults in urban might be higher than those adults in rural. With increasing of age, the plasma TC level should be increased. There is no significant difference in TG levels between the adults aged 45-49 years and over 60 years, while the TG level of both age groups should be higher than those of adults aged 18-44 years. PMID- 16266539 TI - [The prevalence of body overweight and obesity and its changes among Chinese people during 1992 to 2002]. AB - OBJECTIVE: To describe the prevalence of body overweight and obesity and its changes among Chinese people during 1992-2002, and to provide scientific basis for developing intervention strategies for obesity in China. METHODS: The data from "1992 China third National Nutrition Survey" (78,704 subjects, 38,323 male and 40,381 female) and from "2002 China National Nutrition and Health Survey" (209,849 subjects, 101,377 male and 108,472 female) were used in this study. The overweight and obesity were defined by using WHO Z-score criteria for children younger than 7 years old, Chinese age-, sex-specific BMI criteria for children aged 7-17 years, and Chinese BMI criteria for adults. RESULTS: The overall prevalence of overweight and obesity of Chinese people was increased by 38.6% and 80.6%, respectively during the period of 1992-2002. The prevalence of overweight and obesity of Chinese people aged 0-6 years, 7-17 years, 18-44 years, 45-59 years and above 60 years was increased 31.7%, 17.9%, 66.7%, 45.2% and 43.7%, respectively. The increase of the prevalence of overweight and obesity was higher in girls younger than 6 years than their male counterparts, while among other age groups, that were higher in boys than girls. The prevalence of overweight and obesity among people living in rural areas was lower than that of their urban counterparts, while the increment of overweight and obesity prevalence among rural people was greater than that of their urban counterparts. It was estimated that another 70 million overweight and 30 million obese Chinese people emerged in China from 1992 to 2002. CONCLUSION: The prevalence of overweight and obesity of Chinese people was increased rapidly in the past decade, which had affected 260 million Chinese people. It would continue to increase in the near future if effective intervention measures have not been taken. PMID- 16266540 TI - [The current prevalence status of body overweight and obesity in China: data from the China National Nutrition and Health Survey]. AB - OBJECTIVE: To study on the prevalence of body overweight and obesity in China and its distribution in age, sex and economic development status. METHODS: A total of 209,849 in all ages from the 2002 China National Nutrition and Health Survey (NNAHS) database were analyzed. The year 2000 population census data was used for age standardization. 1978 WHO criteria using Z scores was applied to age 0-7 years, criteria recommended by the Working Group on Obesity in China was applied to those aged 7-17 years and criteria in the Guidelines for Prevention and Control of Overweight and Obesity in Chinese Adults was applied to those aged 18 years and above. RESULTS: The overall prevalence was 17.6% for overweight and 5.6% for obesity. The combined prevalence of overweight and obesity was 23.2%. The prevalence of overweight was 25.0%, 21.6%, 17.4%, 15.1%, 19.2% and 12.8%, respectively for large city, middle-and-small city, class 1 rural, class 2 rural, class 3 rural and class 4 rural. For obesity, the rate was 10.6%, 7.2%, 6.4%, 4.3%, 6.0% and 2.7%, respectively for large city, middle-and-small city, class 1 rural, class 2 rural, class 3 rural and class 4 rural. By age groups, the prevalence of overweight was 3.4% in children age 0-6, 4.5% in adolescents age 7 17, 22.8% in adults age 18 and above; the prevalence of obesity was 2.0% in children age 0-6, 2.7% in adolescents age 7-17, 7.1% in adults age 18 and above. The sex difference in overweight and obesity differed between urban and rural, and among age groups. CONCLUSION: Overweight and obesity should have affected nearly 1 quarter of the country's total population, and became a threatening hazard to resident's health. As the urbanization progress, obesity might doubly increase. Prevention and control of this hazard should be urgently needed. PMID- 16266538 TI - [Epidemiologic characteristics of dyslipidemia in people aged 18 years and over in China]. AB - OBJECTIVE: To study the difference in prevalence of dyslipidemia, hypercholesterolemia, hypertriglyceridemia and low blood high-density lipoprotein cholesterol (HDL-C) between the residents of urban and rural areas of varied regions in China. METHODS: Fasting plasma levels of total cholesterol, triglyceride and HDL-C were determined with the enzyme methods for 49,252 subjects aged 18 and over during August to December, 2002. RESULTS: Prevalence of dyslipidemia in Chinese adults aged 18 and over was 18.6%, with 17.0%, 22.9% and 23.4% in the groups of 18-44, 45-59 and over 60 years old, respectively, 22.2% and 15.9% in males and females, respectively, and 21.0% and 17.7% in urban and rural areas, respectively. Prevalence of hypercholesterolemia, hypertriglyceridemia and low blood HDL-C in those aged 18 and over was 2.9%, 11.9% and 7.4%, respectively. CONCLUSIONS: Dyslipidemia has become one of important risk factors threatening health of Chinese people, with hypertriglyceridemia and low blood HDL-C as two major types in those aged 18 and over. Prevalence of dyslipidemia was nearly the same in the middle-aged and in the elderly people, and not significantly different in those living in urban areas from those in rural areas. It is very important to pay more attention to earlier comprehensive prevention and control of dyslipidemia. PMID- 16266542 TI - [An epidemiological study on vaccine derived polio virus circle in Zhenfeng County of Guizhou Province]. AB - OBJECTIVE: To analyze the event and cause of vaccine derived polio virus (VDPV) circle happened at Yaoshang Village, Wanlan Township, Zhenfeng County, Qianxinan Prefecture, Guizhou Province in August 2004. METHODS: Virus isolation was performed for stool specimens collected from two children cases with acute flaccid paralysis (AFP) reported at Yaoshang Village and 21 normal children under five years old in the same village. And, routine immunization coverage at the Village and AFP incidence and performance of AFP surveillance system in Zhenfeng County were investigated, as well. RESULTS: Vaccine derived polio virus 1 (VDPV1) was identified in the feces of two affected children at Yaoshang Village and three other normal children at the same village. Totally, seven cases of AFP have been underreported in Zhenfeng County since 2002. Routine immunization coverage was rather low at Wanlan Township, and nearly nil at Yaoshang Village. CONCLUSIONS: Oral polio vaccine (OPV) with attenuated live virus has been used in China for many years to prevent polio. Circulation of VDPV in Zhenfeng County issues a new challenge to eradication of polio. It is necessary to strengthen routine immunization and AFP surveillance to prevent occurrence of VDPV event. PMID- 16266543 TI - [Effects of selenium and B-27 supplements on viability and differentiation of neural stem cell in newborn rat]. AB - OBJECTIVE: To assess how trace element selenium and B27 supplements affect the neural stem cell (NSc) differentiation in vitro. METHODS: The development and differentiation of NSc from the newborn rat were observed with primary culture and subculture during treating by sodium-selenite, and selenium-methyl-cysteine (SMC). The immunocytochemistry techniques were used to identify the NSc and mature protein expression with neuron marker beta-tubulin, astrocyte marker GFAP, and oligodendrocyte marker CNPase. The neurosphere morphology and neurite outgrowth were observed. RESULTS: Adding the complete B-27 serum-free supplement, Selenium could promote the neurosphere viability, development and differentiation. Without selenium and B-27, neurosphere could not survive and differentiate. Without B-27 in the medium but there containing selenium, the neurosphere could promote the viability and development into neuron, astrocyte and oligodendrocyte, as compared with the no-containing B-27 and selenium groups, these differentiated cells might have more quantity, more branches and better morphological nerve net. The count of the neuron, astrocyte and oligodendrocyte was 11.2/Hp, 16.1/Hp and 9.3/Hp. CONCLUSIONS: The selenium should be very important for neural stem cells' survival. Selenium could promote the neurosphere cells differentiation and development. PMID- 16266544 TI - [Effects of zearalenone on proliferation and apoptosis in MCF-7 cells]. AB - OBJECTIVE: To explore the effects of zearalenone (ZEA) on proliferation and apoptosis in estrogen-dependent human breast cancer MCF-7 cells and the likely underlying molecular mechanisms. METHODS: Cell viability was determined by MTT assay and cell cycle distribution by cytometry. Apoptosis was detected by Cell Death Detection ELISA and cytometry, respectively. The expressions of bax and bcl 2 were examined using multiple RT-PCR and Western-blot both at mRNA and protein level, respectively. RESULTS: The current study confirmed the previous studies that ZEA could stimulate proliferation in MCF-7 cells with inducing a profound increase in S phase and a modest increase in G(2)/M phase that was accompanied by a decrease in G(0)/G(1) phase. ZEA could inhibit apoptosis in MCF-7 cells following estrogen ablation at a range of concentrations of 2 nmol/L -96 nmol/L. Western blot and RT-PCR analysis revealed that the anti-apoptotic bcl-2 was upregulated at both protein and mRNA level, together with the downregulation of pro-apoptotic bax. CONCLUSION: ZEA should have possessed comparative estrogenic activity and could promote the progression of MCF-7 cells through the cell cycle by a decreasing in the G(0)/G(1) phase and by a significant increasing in S phase. The pro-proliferative activity of ZEA was due to inhibition of apoptosis through regulation of bax/bcl-2 expression. PMID- 16266545 TI - [Analysis of hepatitis virus infection in hepatocellular carcinoma patients receiving surgical operation in China and Japan]. AB - OBJECTIVE: To describe and compare the hepatitis virus infection in hepatocellular carcinoma (HCC) patients receiving surgical operation in China and Japan. METHODS: Information of surgical HCC patients was retrieved from the medical records. The concerned characteristics of the HCC cases from two countries were described and compared. RESULTS: A total of 425 diagnosed cases that underwent surgical resection for HCC in China were investigated, and the corresponding cases in Japan were 247. The proportion of the hepatitis virus infection was 75.53% in patients with HCC from China. Within the infection cases, 91.28% were positive for HBsAg but negative for anti-HCV. The proportion was 82.59% in patients with HCC from Japan. Within the infection cases, 77.94% were positive for anti-HCV but negative for HBsAg. The proportion of hepatocirrhosis in the hepatitis virus infection patients with HCC were 89.10% and 68.14% in China and Japan, respectively. CONCLUSION: The hepatitis B virus infection showed be a main cause of HCC in China, however, the HCC in Japan be mostly related to hepatitis C virus infection. PMID- 16266547 TI - [Investigation of dietary fiber intakes and varies in 53 patients with diabetes]. AB - OBJECTIVE: To investigate the intakes of total dietary fiber (TDF), soluble dietary fiber (SDF) and insoluble dietary fiber (IDF) in Subjects With Type 2 Diabetes so as to provide the base for making the adequate intakes of dietary fiber. METHODS: The enzymatic-gravimetric methods for dietary fiber were established on basis of a collaborative study. Dietary intake was measured by means of 3-day food records through weighting and using food pictures. TDF, SDF and IDF were analyzed by enzymatic-gravimetric methods. RESULTS: The reproducibility relative standard deviations for DF ranged from 2.63% to 9.67%. Vegetable foods were the mainly sources of DF. The total dietary intakes, insoluble and soluble fibers were 26.5 +/- 9.8, 14.6 +/- 5.8, 10.4 +/- 4.4 (g/d) respectively. CONCLUSION: The dietary fiber intake of the diabetes remains in the range of intakes recommended by American Diabetes Association. PMID- 16266546 TI - [Application of flow cytometry on functional assessment of health food]. AB - OBJECTIVE: To establish flow cytometry (FCM) methods and evaluate their application value for measuring the index for enhancing immune function of health food. METHODS: In mice experiment model, the dosage groups were respectively oral fed with three test substances according to 5, 10, 30 times of the recommended dose for human body; both the negative and positive control groups were fed with equivalence purified water once a day. The positive control was fed with 25 mg/kg body weight levamisole for 3 days before finishing the administration, and the immune two percent of sheep erythrocytes were administrated at the last day. In rats experiment model, the test substance was given by mixing feed according to 25 and 50 times of the recommended dose for human body. At the end of the experiment, indices below were simultaneously detected. (1) The classical indices included: spleen lymphocyte transformation test by using ConA (MTT assay); spleen NK cell activity test (LDH assay); delayed-type hypersensitivity test by using sheep erythrocyte (foot palm thickening) method and phagocytosis activity tested by mice peritoneal macrophages. (2) FCM indices included: T and B lymphocytes quantitating in mice peripheral blood, activated antigen expression level in the surface of T lymphocytes and NK cells and phagocytosis activity for fluospheres in mice peritoneal macrophages. RESULTS: (1) Compared with the negative control group, there were no significant differences in T and B lymphocytes proportion and the number of lymphocytes in mice peripheral blood after given 0.83, 1.67, 5.01 g/kg protein powder; (2) mice peripheral blood T lymphocyte sub-cluster CD(69)(+)/CD(3)(+) of 3.75, 7.50, 15.0 ml/kg bw Cen-Rong Cream groups were all significantly increased (P < 0.05), which were shown a good coherence with the classic test index; (3) mice peripheral blood NK cell sub-cluster CD(69)(+)/NKG2D(+) of 0.83, 1.67 g/kg protein powder groups were both significantly increased (P < 0.05), which was kept in good coherence with those of NK cell activity test (LDH assay); rats peripheral blood NK cell sub-cluster CD(161a+)/CD(25)(+) of 1.50 g/kg ganoderma lucidum and cordycepicmycelia group was significantly increased (P < 0.05); (4) the phagocytosis activity in mice peritoneal macrophages: there were no significant difference found between the controls and the dosage groups in the classic test. However, in the FCM test, the percentage of phagocytic cells of 0.15, 0.30, 0.90 g/kg ganoderma lucidum and cordycepicmycelia groups and the phagocytic index of 0.30, 0.90 g/kg were enhanced. CONCLUSION: It suggests that was shown in detecting and assessing enhancing immune function of health food the results tested by FCM were fairly consistent with those by using traditional methods, most of them would have higher sensitivity. It should be valuable to applying FCM in the measurement and assessment of enhancing immune function of health food and worth while to further study as to enlarging its application. PMID- 16266548 TI - [Genetic effects on physical growth items of child twins in Han nationality]. AB - OBJECTIVE: To analyse the heritabilities of physical growth items of body and its related factors. METHODS: An 116 twin pairs of Han nationality, 67 monozygotic (MZ) and 49 like-sex dizygotic (DZ) aged 6 to 12 years, were investigated from June to October in 2004. The measurements included height, weight, sitting height, chest circumference, biacromial breadth and biiliac breadth, and BMI index calculated by the former two measurements. The heritabilities were estimated by using intraclass correlation coefficient method from the adjusted data for age. RESULTS: The intraclass correlation coefficient was greater in the MZ twins than in the DZ twins. The estimated heritabilities of height, weight, BMI, sitting height, chest circumference, biacromial breadth and biiliac breadth were 0.89, 0.88, 0.73, 0.87, 0.78, 0.78, 0.73 in boys and 0.87, 0.74, 0.72, 0.86, 0.62, 0.56, 0.59 in girls adjusted for age. Therefore, there were no sex difference for the heritabilities of height, sitting height and BMI, but the male heritabilities of weight, chest circumference, biacromial breadth and biiliac breadth were higher than the female's respectively. CONCLUSION: Physical growth items should be mainly determined by the genetic factors. There are sex differences for the heritabilities of weight, chest circumference, biacromial breadth and biiliac breadth, i.e., the girls might be affected more easily by environmental factors than the boys in these items. PMID- 16266549 TI - [Anxiety state and its related factors in Shanghai high school students]. AB - OBJECTIVE: To investigate the anxiety state in high school students and related factors in order to get reasonable suggestions for prevention. METHODS: The mental health test (MHT) for high school students, and the living environmental and parental style were used in this study. RESULTS: MHT served as an assessment scale of anxiety. The efficiency sample was 3,050, aged 11 to 18 years old. The level of total anxiety and its each contents was low to moderate (0.24 to 0.54). The percentage of moderate to high of total anxiety was 16.7%, the percentage of moderate to high of each anxiety aspects were 8.8%-21.8%. The mostly high aspects were self-blame, schooling anxiety, social anxiety and over sensitiveness. In general, the girls' anxiety level was higher than boys', but the boys' lonely feeling was higher than the girls'. The total score of anxiety was decreased with age. Except of the over sensitiveness, the decrease tendency of each anxiety contents was significant in boys. For girls, the lonely feeling was deceased and the over sensitiveness increased with age. The age of fifteen seems as a significant changing age. The related disadvantage factors of students' anxiety were: the low education level, the parents' anxiety and depression characters, the authoritarian or neglecting parental style, the often contradiction parental styles between mother and father, parents often quarrel, the experience of often being scalded and physical punishment, lacking in care of others when in difficulties. CONCLUSIONS: The schooling pressure should be decreased to an appropriate level. The students' self-confidence and social ability should be emphasized. Good family environments and the support outside the family should be quite important for adolescents' mental health, these factors might decrease the adolescents' anxiety. PMID- 16266560 TI - The effectiveness and cost-effectiveness of dual-chamber pacemakers compared with single-chamber pacemakers for bradycardia due to atrioventricular block or sick sinus syndrome: systematic review and economic evaluation. AB - OBJECTIVES: To estimate the effectiveness and cost-effectiveness of dual-chamber pacemakers versus single-chamber atrial or single-chamber ventricular pacemakers in the treatment of bradycardia due to sick sinus syndrome (SSS) or atrioventricular block (AVB). DATA SOURCES: Electronic databases and relevant Internet sites. Contact with device manufacturers and experts in the field. REVIEW METHODS: A systematic review was carried out of randomised controlled trials (RCTs). The quality of selected studies was appraised using standard frameworks. Meta-analyses, using random effects models, were carried out where appropriate. Limited exploration of heterogeneity was possible. Critical appraisal of economic evaluations was carried out using two frameworks. A decision-analytic model was developed using a Markov approach, to estimate the cost-effectiveness of dual-chamber versus ventricular or atrial pacing over 5 and 10 years as cost per quality-adjusted life-year (QALY). Uncertainty was explored using one-way and probabilistic sensitivity analyses. RESULTS: The searches retrieved a systematic review of effectiveness and cost-effectiveness published in 2002, four parallel group RCTs and 28 cross-over trials. Dual-chamber pacing was associated with lower rates of atrial fibrillation, particularly in SSS, than ventricular pacing, and prevents pacemaker syndrome. Higher rates of atrial fibrillation were seen with dual-chamber pacing than with atrial pacing. Complications occurred more frequently in dual-chamber pacemaker insertion. The cost of a dual-chamber system, over 5 years, including cost of complications and subsequent clinical events in the population, was estimated to be around 7400 pounds. The overall cost difference between single and dual systems is not large over this period: around 700 pounds more for dual-chamber devices. The cost effectiveness of dual-chamber compared with ventricular pacing was estimated to be around 8500 pounds per QALY in AVB and 9500 pounds in SSS over 5 years, and around 5500 pounds per QALY in both populations over 10 years. Under more conservative assumptions, the cost-effectiveness of dual-chamber pacing is around 30,000 pounds per QALY. The probabilistic sensitivity analysis showed that, under the base-case assumptions, dual-chamber pacing is likely to be considered cost effective at levels of willingness to pay that are generally considered acceptable by policy makers. In contrast, atrial pacing may be cost-effective compared with dual-chamber pacing. CONCLUSIONS: Dual-chamber pacing results in small but potentially important benefits in populations with SSS and/or AVB compared with ventricular pacemakers. Pacemaker syndrome is a crucial factor in determining cost-effectiveness; however, difficulties in standardising diagnosis and measurement of severity make it difficult to quantify. Dual-chamber pacing is in common usage in the UK. Recipients are more likely to be younger. Insufficient evidence is currently available to inform policy on specific groups who may benefit most from pacing with dual-chamber devices. Further important research is underway. Outstanding research priorities include the economic evaluation of UKPACE studies of the classification, diagnosis and utility associated with pacemaker syndrome and evidence on the effectiveness of pacemakers in children. PMID- 16266565 TI - [Organization of colorectal surgery in Denmark--on which basis?]. PMID- 16266559 TI - Long-term outcome of cognitive behaviour therapy clinical trials in central Scotland. AB - OBJECTIVES: To establish the long-term outcome of participants in clinical trials of cognitive behaviour therapy (CBT) for anxiety disorders and psychosis, examining the effectiveness and cost-effectiveness associated with receiving CBT in comparison with alternative treatments. DESIGN: An attempt was made to contact and interview all of the participants in eight randomised, controlled, clinical trials of CBT for anxiety disorders and two randomised, controlled, clinical trials of CBT for schizophrenia conducted between 1985 and 2001. Case note reviews of healthcare resources used in the 2 years prior to entering the trials and the 2 years prior to follow-up interview were undertaken. SETTING: Mixed rural and urban settings in five localities in central Scotland. Anxiety disorder trials were conducted mainly in primary care and included three with generalised anxiety disorder, four with panic disorder and one with post-traumatic stress disorder (PTSD). The psychosis studies (one on relapse prevention and one with chronic disorder) were conducted in secondary care. PARTICIPANTS: Of the 1071 entrants to the 10 studies, 489 agreed to participate (46% of original entrants, 52% of those available to contact). INTERVENTIONS: Follow-up interviews took place between 1999 and 2003, 2-14 years after the original treatment. Interviews for Trials 1-8 were conducted by a research psychologist blind to original treatment condition. Interviews for Trials 9 and 10 were conducted by community psychiatric nurses also blind to treatment condition. Case note reviews were completed following the interview. MAIN OUTCOME MEASURES: For Trials 1-8 the main interview-based outcome measures were: Anxiety Disorders Interview Schedule-DSM IV for diagnosis and co-morbidity, Clinical Global Severity (0-8) and the Hamilton Anxiety Rating Scale. The main patient-rated measures were: Brief Symptom Inventory, SF-36 II, Clinical Global Improvement (1-7), and the Positive and Negative Affect Scale. For Trials 9 and 10 the primary outcome measure was the interview-based Positive and Negative Syndrome Scale (PANSS). RESULTS: For the anxiety disorder studies (Trials 1-8), over half of the participants (52%) had at least one diagnosis at long-term follow-up, with significant levels of co morbidity and health status scores comparable to the lowest 10% of the general population. Only 36% reported receiving no interim treatment for anxiety over the follow-up period with 19% receiving almost constant treatment. Patients with PTSD did particularly poorly. There was a 40% real increase in healthcare costs over the two time periods, mainly due to an increase in prescribing. A close relationship was found between poor mental and physical health for those with a chronic anxiety disorder. Treatment with CBT was associated with a better long term outcome than non-CBT in terms of overall symptom severity but not with regard to diagnostic status. The positive effects of CBT found in the original trials were eroded over longer time periods. No evidence was found for an association between more intensive therapy and more enduring effects of CBT. Long term outcome was found to be most strongly predicted by the complexity and severity of presenting problems at the time of referral, by completion of treatment irrespective of modality and by the amount of interim treatment during the follow-up period. The quality of the therapeutic alliance, measured in two of the studies, was not related to long-term outcome but was related to short-term outcome. The cost-effectiveness analysis showed no advantages of CBT over non CBT. The cost of providing CBT in the original trials was only a very small proportion (6.4%) of the overall costs of healthcare for this population, which are high for both physical and mental health problems. In the psychosis studies (Trials 9 and 10), outcome was generally poor with only 10% achieving a 25% reduction in total PANSS scores from pretreatment to long-term follow-up, also cost-effectiveness analysis showed no advantages of CBT over non-CBT, although healthcare costs fell over the two time periods mainly owing to a reduction in inpatient costs. CONCLUSIONS: Psychological therapy services need to recognise that anxiety disorders tend to follow a chronic course and that good outcomes with CBT over the short term are no guarantee of good outcomes over the longer term. Clinicians who go beyond standard treatment protocols of about 10 sessions over a 6-month period are unlikely to bring about greater improvement. Poor outcomes over the long term are related to greater complexity and severity of presenting problems at the time of referral, failure to complete treatment irrespective of modality and the amount of interim treatment during the follow-up period. The relative gains of CBT are greater in anxiety disorders than in psychosis. Longitudinal research designs over extended periods of time (2-5 years), with large numbers of participants (500+), are required to investigate the relative importance of patient characteristics, therapeutic alliance and therapist expertise in determining the cost-effectiveness of CBT in the longer term. PMID- 16266566 TI - [Colorectal cancer. Incidence and risk factors]. AB - Colorectal cancer (CRC) is a common cancer in the more developed Western countries. This paper aims to give a review of the established or suspected and potentially modifiable risk factors for CRC. Increased BMI, physical inactivity, tobacco smoking and high alcohol consumption are strongly implicated factors. The precise dietary patterns associated with increased risk are less clear. Use of NSAIDs as well as postmenopausal hormones seems to be associated with a substantially reduced risk of CRC. Altogether, the evidence indicates that primary prevention of CRC is feasible. PMID- 16266567 TI - [Hereditary colorectal cancer]. AB - HNPCC and FAP are inherited diseases with a lifetime risk of colorectal cancer (CRC) of 80-100% in gene carriers. Disease-causing mutations have been identified in the APC gene at FAP and in MMR genes at HNPCC. In FAP-patients, screening has reduced the prevalence of CRC by 55%, and the survival rate has improved considerably. For HNPCC-patients, 77% of CRCs found by screening were Duke' A or B, and survival after CRC has improved significantly since 1990. Continuous central registration in the HNPCC and Polyposis registers is recommended to ensure identification of high-risk families and evaluate the effect of screening. PMID- 16266568 TI - [Serological markers of colorectal cancer]. AB - In Denmark, approximately 3,600 persons are diagnosed with colorectal cancer every year. Of these patients, 75-80% may undergo intended curative resection, but within a few years the disease recurs in about half of the curatively resected patients. Less than half of the total number of patients will survive the subsequent five years. Radiotherapy and adjuvant systemic chemotherapy have a limited effect on overall survival. It is therefore imperative to optimise the overall treatment of patients with colorectal cancer. One substantial option might be the development, validation and implementation of biological markers that could be used for early detection/screening, prognosis evaluation, surveillance, selection of patients for adjuvant therapy independent of stage of the disease, and monitoring of the given treatment. PMID- 16266569 TI - [Immunohistochemical markers in colorectal cancer]. AB - In colorectal cancer, immunohistochemical staining can routinely be used for tumour classification, screening for defective DNA mismatch repair and possibly identification of markers relevant to prediction of outcome of chemotherapy. This article provides a brief description of these applications. PMID- 16266570 TI - [Quality control of pathological examination in colorectal cancer]. AB - Pathological assessment of colorectal cancer resection specimens is the most significant prognostic indicator. This includes determination of TNM stage, tumour type and grade, status of resection margins, extramural vascular and perineural invasion, and molecular features such as defects in the mismatch repair system. The pathological assessment contributes significantly to the quality control of surgery and preoperative staging. Reporting should be standardized by using DSPAC's registration scheme for colorectal carcinomas. PMID- 16266571 TI - [Diagnostic imaging in staging of colorectal cancer]. AB - The main strength of MR is the ability to define the mesorectal facia. There is recent evidence suggesting that MR imaging can accurately predict the tumor-free circumferential resection margin before total mesorectal excision. However, the prediction of T stage is less accurate and more affected by the experience of the observer. Transrectal ultrasonography has proven useful for staging of rectal carcinoma. Computed tomography is one of the most useful imaging techniques for the evaluation of metastatic lesions in colorectal cancer. PMID- 16266572 TI - [Virtual colonoscopy is now reality]. AB - Virtual colonoscopy involves a helical CT or MR scan of the abdomen and pelvis to detect colorectal polyps and cancer. Both modalities have shown promising sensitivity in revealing larger polyps, in comparison with colonoscopy. Caution should be exercised in its clinical implementation due to significant interobserver variation and individual learning curves. A Danish study indicates that CT colonography (CTC) can be performed cost-effectively compared to colonoscopy. CTC is recommended in preference to double-contrast barium enema after incomplete colonoscopy. PMID- 16266573 TI - [Diagnostic strategy with respect to symptoms of colorectal cancer]. AB - In relation to the risk of colorectal cancer, the population is divided into high risk and average-risk groups. Patients at high risk are followed according to the national recommendations. Patients at average risk are examined according to their primary symptom(s). The strategy is based on the diagnostic strategy described in a Danish Health Technology Assessment published in 2001. The article states the recommendations for the high- risk groups and describes the value of the individual primary symptoms of colorectal cancer for average-risk patients and the examinations recommended accordingly. PMID- 16266574 TI - [Acute treatment of colon cancer]. AB - The primary treatment of left-sided colonic ileus due to cancer is after localisation of the tumor with water-soluble contrast enema placement of an intraluminal, self-expanding metal stent. If this treatment is unsuccessful, open resection with primary anastomosis is the treatment of choice. In the case of a perforated tumor, resection and primary anastomis may be performed if the patient s general condition is acceptable. PMID- 16266575 TI - [Fast-track colorectal cancer surgery]. AB - Multimodal rehabilitation with preoperative information, reduction of surgical stress responses, optimised dynamic pain relief and early mobilisation and oral nutrition reduce hospital stay, morbidity and convalescence in colorectal procedures. In addition, the results of fast-track colonic surgery suggest that postoperative pulmonary, cardiovascular and muscle function are improved and body composition preserved. Consequently, the hospital stay may be reduced to about two to four days. Preliminary studies suggest similar improvements in more complex colorectal procedures. PMID- 16266576 TI - [The Danish Colorectal Cancer Database]. AB - The Danish Colorectal Cancer Database was established in 1994 with the purpose of monitoring whether diagnostic and surgical principles specified in the evidence based national guidelines of good clinical practice were followed. Twelve clinical indicators have been listed by the Danish Colorectal Cancer Group, and the performance of each hospital surgical department with respect to these indicators is reported annually. In addition, the register contains a large collection of data that provide valuable information on the influence of comorbidity and lifestyle factors on disease outcome and survival. PMID- 16266577 TI - [Follow-up of patients after curative surgery for colorectal cancer]. AB - Meta-analyses of randomised controlled studies show that intensive follow-up programmes after radical surgery for colorectal cancer have a more positive effect on total survival than do less intensive programmes, but the ideal programme has not yet been found. It appears that monitoring with carcinoembryonal antigen (CEA) and some kind of visualisation of the liver is a way forward, but we do not know how often it should be done. Colonoscopy should be offered every five years to look for metachronous cancer; patients with HNPCC should have a colonoscopy done even more often. PMID- 16266578 TI - [Future organisation of colorectal cancer surgery in Denmark]. AB - We recommend that in the future, surgery for colorectal cancer (CRC) in Denmark should be done in 10 to 15 colorectal units with an uptake zone of 350,000 500,000 citizens each. These units should perform both acute and elective CRC surgery and acute surgical treatment of other intestinal diseases. In each unit, a senior colorectal surgeon should be available on a 24-hour shift, and there should be sufficient diagnostic and theatre capacity to ensure optimal treatment levels. A stoma clinic should be available Monday to Friday. The units should perform research according to international standards. Each senior surgeon should document his or her relevant continuing surgical education. A national postgraduate education should be ensured to all members of the multidisciplinary team according to the standards in force in Great Britain. The treatment of primary advanced T4 tumors and local recurrence should be done in only one to two colorectal units in Denmark, and the treatment of local recurrence with spread to the pelvis should be done in only one unit in Denmark. PMID- 16266579 TI - [Future perspectives in screening for colorectal cancer]. AB - Based on international experience, pilot studies with fecal occult blood tests and colonoscopy in those with blood in the stools are now to be performed in Scandinavia in limited geographic areas before introducing countrywide screening. The Danish State has offered to pay for feasibility studies in two counties. Provided that the results of randomised clinical trials are reproduced according to acceptability and stage of the cancer found and the organisation is feasible, it will then be possible to evaluate newer technologies as well as other methods which have not previously been evaluated in a satisfactory way. PMID- 16266584 TI - [TIPS is a life-prolonging treatment of those cirrhosis patients with ascites who can not be treated medically or with paracentesis]. PMID- 16266586 TI - [Documentation of acupuncture]. PMID- 16266589 TI - [Physical activity increases the quality of life]. PMID- 16266599 TI - A young girl with scaly skin plaques. PMID- 16266598 TI - Vaginal pain and fever in a premenarchal girl. How would you treat? PMID- 16266600 TI - The uninsured: you can make a difference. PMID- 16266601 TI - Chronic compartment syndrome: tips on recognizing and treating. PMID- 16266602 TI - Acute otitis media: influence of the PCV-7 vaccine on changes in the disease and its management. AB - Studies have shown that most cases of acute otitis media (AOM) are of viral or "no bacteria isolated" origin. In these cases, watchful waiting may be appropriate. This publication addresses presumed bacterial causes of AOM for which antibiotic use may be efficacious. PMID- 16266603 TI - Is your practice really that predictable? Nonlinearity principles in family medicine. PMID- 16266604 TI - No need for glycosuria/proteinuria screen in pregnant women. AB - OBJECTIVE: More than 22 million prenatal visits occur in the US each year. Each pregnant woman averages 7 visits. Most include urine testing for glucose and protein to screen for gestational diabetes and preeclampsia. Is there sufficient scientific evidence to support this routine practice? METHODS: We searched Medline (1966-2004), the Cochrane review, AHRQ National Guideline Clearinghouse, the Institute for Clinical Systems Improvement, and Google, searching for studies on proteinuria or glycosuria in pregnancy. The reference list of each article reviewed was examined for additional studies, but none were identified. We found 6 studies investigating glycosuria as a predictor for gestational diabetes mellitus, or proteinuria as a predictor for preeclampsia (1 examined both). Because every study used different dipstick methods of determining results, or definitions of abnormal, each was evaluated separately. RESULTS: Glycosuria is found at some point in about 50% of pregnant women; it is believed to be due to an increased glomerular filtration rate. The renal threshold for glucose is highly variable and may lead to a positive test result for glycosuria despite normal blood sugar. High intake of ascorbic acid or high urinary ketone levels may result in false-positive results. Four published studies assessed the value of glycosuria as a screen for gestational diabetes. All used urine dipsticks. Three of the 4 most likely overestimate the sensitivity of glycosuria for predicting gestational diabetes. CONCLUSIONS: Routine dipstick screening for protein and glucose at each prenatal visit should be abandoned. Women who are known or perceived to be at high risk for gestational diabetes or preeclampsia should continue to be monitored closely at the discretion of their clinician. PMID- 16266605 TI - Which late-stage Alzheimer's patients should be referred for hospice care? PMID- 16266606 TI - Clinical inquiries. When is neuroimaging warranted for headache? PMID- 16266607 TI - Clinical Inquiries. What are the best therapies for acute migraine in pregnancy? PMID- 16266608 TI - Clinical Inquiries. How does colonoscopy compare with fecal occult blood testing as a screening tool for colon cancer? PMID- 16266609 TI - Clinical Inquiries. How can we best treat and monitor VTE during pregnancy? PMID- 16266610 TI - What is the best way to diagnose menopause? PMID- 16266611 TI - [Infectious endocarditis]. PMID- 16266612 TI - [Minimal residual disease in acute lymphoblastic leukemia: a new concept of complete remission]. AB - INTRODUCTION: Early response to induction treatment is one of the most important prognostic factors in children with acute lymphoblastic leukemia (ALL). Cytological remission is currently achieved in 95-98 % of these patients, although a significant proportion will later relapse. More sensitive techniques are required to measure residual leukemia and establish a new definition of complete remission. OBJECTIVE: To identify minimal residual disease (MRD) by immunological techniques and define its prognostic impact in children with ALL. METHODS: MRD was studied by flow cytometry in 53 children diagnosed in our department between June 1999 and April 2003 and treated using the Pethema protocols. All the children achieved complete cytological remission (< 5 %) with the induction treatment and had at least one useful phenotype for follow-up: 11 % were T phenotype, one was biphenotypic and the remainder were B cell leukemias. Bone marrow samples were analyzed post-induction, post-consolidation, after 6 and 11 months of maintenance treatment, at the end of treatment, and 3 months later. The positivity threshold was set at 0.01 % and the sensitivity of the technique was 1 x 10(-4)-1 x 10(-5). RESULTS: A total of 199 samples were analyzed. Thirty seven percent of the post-induction and 20 % of the post-consolidation samples analyzed were MRD-positive. Elimination was slower in patients with a T phenotype and in high-risk patients according to the traditional classification. After a median follow-up of 26 months, event free survival (EFS) in the group as a whole was 92 %. The EFS rate in the patients who were MRD-positive post-induction was 79 %. None of the patients who were MRD-negative post-induction has developed recurrence. CONCLUSION: Study of MRD is essential and should be included in all current treatment protocols for children with ALL. Its usefulness derives from the prognostic impact of the response to induction treatment. Continued sequential monitoring may predict recurrence before the onset of clinical or hematologic manifestations. PMID- 16266613 TI - [New trends in pediatric endocarditis]. AB - OBJECTIVES: To evaluate the epidemiology, etiology and outcome of endocarditis in a cohort of pediatric patients and to compare the main characteristics with our previous experience. MATERIAL AND METHODS: Patients aged less than 18 years of age diagnosed with endocarditis at the Sainte-Justine Hospital, University of Montreal between 1-1986 and 12-2000 were studied. The recent case series was compared with our previous experience from 1960-1985. RESULTS: Fifty-six children with endocarditis were included in the 1986-2000 series: 35 children with congenital heart disease, 15 with serious underlying disease and six healthy children. The mean age was 7 years and 10 months. Fifty-four percent of the patients were boys. The incidence of endocarditis increased from 1.5 cases/year in the 1986-2000 series to 4 cases/year in the 1986-2000 series. In the 1986-2000 series, 10 (17.9 %) patients had a central catheter. Sixteen (28.6 %) patients had a vascular prosthesis. Blood cultures were positive in 50 patients (89 %): Streptococci were found in 48 % of the patients and Staphylococci in 34 %. Echocardiography was positive in 36 of 55 patients (65.4 %). All children were treated with intravenous antibiotics for a mean of 43 +/- 15 days. There were no recurrences. Significant morbidity developed in 26 patients (46 %). Embolic phenomena were seen in 11 children (20 %). Twelve patients (21 %) needed surgery. Of the six healthy children, five developed complications. Overall, seven children (12.5 %) died; all were older than 6 years of age. Comparing our experience from 1960-1985 with that from 1986-2000 revealed that morbidity decreased from 85.7 % to 46.4 % and mortality decreased from 27 % to 12.5 %. CONCLUSIONS: Physicians must recognize that children with underlying immunodeficiency and those with central catheters have an increased risk of endocarditis. Healthy children with endocarditis have a greater risk of complications. Morbidity and mortality due to endocarditis has diminished considerably in recent years. PMID- 16266614 TI - [Salmonellosis in a maternity-children's hospital in Barcelona over a 10-year period (1992-2001)]. AB - BACKGROUND: We performed a retrospective study of Salmonella spp. strains, as well as of serotypes and resistance to antimicrobial agents that could be useful in salmonellosis requiring antibiotic treatment. MATERIAL AND METHODS: All Salmonella strains isolated in a maternity-children's hospital in Barcelona (Spain) during a 10-year period (1992-2001) were serotyped. The susceptibility of the strains isolated from 1994 to ampicillin, amoxicillin-clavulanic acid, chloramphenicol, trimethoprim-sulfamethoxazole and cefotaxime was analyzed. Ciprofloxacin substituted chloramphenicol in the analysis of strains isolated during the last 2 years of the study period. RESULTS AND DISCUSSION: A total of 860 isolates from 31 distinct serotypes were analyzed. The most frequent serotypes were S. enteritidis (48.7 %), S. typhimurium (33.4 %), S. virchow (5 %), S. hadar (1.8 %) and S. typhi (1.5 %). We found a high percentage of strains resistant to ampicillin (44.6 % in 2001) and chloramphenicol (28.7 % in 1999) and a lower percentage of strains resistant to amoxicillin-clavulanic acid and trimethoprim-sulfamethoxazole (2.2 % and 6.5 % respectively, among the strains isolated in 2001). Only one cefotaxime-resistant strain was found, and all isolates were susceptible to ciprofloxacin. Our results provide useful epidemiological information for the control of these infections, which remain a serious public health problem all over the world. PMID- 16266615 TI - [Underlying causes of recurrent pneumonia]. AB - OBJECTIVE: To determine the relative frequency and describe the predisposing causes of recurrent pneumonia in infants and children aged between 1 month and 14 years. METHODS: We retrospectively reviewed the medical records of a tertiary care pediatric hospital covering a 10-year period, from January 1994 through December 2003. Children with cystic fibrosis were not included in the analysis. Recurrent pneumonia was defined as at least two pneumonia episodes in a 1 year period or at least three episodes over a lifetime. RESULTS: Of 1644 children hospitalized with pneumonia, 106 (6.4 %) met the criteria for recurrent pneumonia. An underlying cause was identified in 92 patients (86.7 %). Of these, the underlying cause was diagnosed prior to pneumonia in 67 (72.8 %), during the first episode in 12 (13 %) and during recurrence in 13 (14.1 %). Underlying causes included asthma in 28 patients (30.4 %), congenital cardiac defects in 27 patients (29.3 %), aspiration syndrome in 25 patients (27.1 %), immune disorder in nine patients (9.7 %), pulmonary anomalies in two patients (2.1 %), and anhidrotic ectodermal dysplasia in one patient (1 %). CONCLUSIONS: Recurrent pneumonia occurred in 6.4 % of all children hospitalized for pneumonia. The underlying cause was identified in 86.7 % of the children. The most common causes were asthma, congenital cardiac defects, and aspiration syndrome. PMID- 16266616 TI - [Neonatal hospitalization through a hospital emergency service in Uruguay]. AB - OBJECTIVE: To determine the prevalence of neonatal admissions through a pediatric emergency department (PED), and to evaluate the characteristics of patients who present vital risk on arrival. METHODS: We performed a descriptive, retrospective review of neonates aged 2-28 days arriving from home who presented to the pediatric emergency department between 01/01/03 and 31/12/03. RESULTS: Of a total of 943 neonatal consultations, 277 (29.3 %) required admission. The most common diseases were respiratory (42.6 %) and fever without source (17 %). Among the 943 patients attended, 42 (4,5 %) showed vital risk, and in this group, 81 % were full term neonates, with a P50 of age of 19 days, and a P50 of weight of 3200 g. Among these, there was a predominance of infectious diseases (74 %), especially respiratory tract infections (50 %). Respiratory syncytial virus (RSV) was identified in 14 of the 42 patients. Thirty-six percent required mechanical ventilation and 71.5 % were transferred to the pediatric intensive care unit. Sixty percent of the critically-ill neonates were admitted in the 3 winter months (p < 0.001). CONCLUSIONS: A high percentage of the neonates arriving from home required admission, and vital risk was more frequent in this age group than in other ages (p < 0.001). This does not correspond to risk related to perinatal pathology, or to nutritional reasons, but to community-acquired infections, particularly those of the respiratory tract, the most frequent cause of which is RSV. PMID- 16266618 TI - [Ondine syndrome: diagnosis and management]. AB - Ondine's syndrome is a congenital central hypoventilation syndrome due to a disorder in the autonomic control of breathing in the absence of any primary disease that would explain it. Although the incidence of this entity is low, it may be underestimated due to the variable clinical expression of this syndrome, depending on its severity. Early diagnosis is of great importance to provide appropriate management to prevent the acute and chronic asphyxia that determines the long-term prognosis of this disease. This review aims to present practical management guidelines to provide doctors unfamiliar with this syndrome with basic knowledge of the diagnosis, treatment and follow-up of these patients. PMID- 16266617 TI - [Functional constipation: prospective study and treatment response]. AB - OBJECTIVE: To assess the prevalence of functional constipation (FC) with and without encopresis, the factors involved in its onset, and treatment response. PATIENTS AND METHODS: A prospective study of 62 children was performed using a standard questionnaire (onset-age, regular toilet use, encopresis, complications, dietary habits and environmental and psychological factors) and physical and anthropometric assessment. FC was defined as a stool frequency of less than 3 bowel movements/week, with passage of large or scybalous stools, with or without 2 or more soiling episodes per week, without underlying disease. Treatment included demystification, behavioral modification and drugs (mineral oil and senna). Each child was periodically re-evaluated, and treatment was considered successful when the defecation rate was 3 or more bowel movements/week, discomfort was absent, and fecal soiling frequency was 2 or fewer episodes/ month. RESULTS AND CONCLUSIONS: FC accounted for 13 % of all first consultations (60 % boys, 40 % girls; mean age at diagnosis 6.1 years). The most frequent manifestations were painful defecation (60 %), rectorrhagia (42 %), obstructive episodes (34 %) and anal fissure or hemorrhoids (17 %); 19 patients (31 %) had encopresis. Nutritional assessment revealed that 84 % of the patients was well nourished and 16 % was overweight. Fiber intake was deficient in more than 60 %. Sixteen (26 %) patients underwent successful relief of impaction with senna (20 30 mg/dose) combined with mineral oil. Maintenance treatment included mineral oil (15-30 ml/day) and senna at the minimum effective dose (5-15 mg/day). Satisfactory results were achieved 1 month later in 32 % of the children, 3-6 months later in 71 %, and 6-12 months later in 85 %; successful response was closely related to regular toilet habits, dietary modification and a shift in the family's attitude. PMID- 16266619 TI - [The cannabinoid system and its importance in the perinatal period]. AB - The cannabinoid system has been recently described, including the endogenous ligands, mainly arachidonic acid derivatives, and their specific receptors. Endocannabinoids are involved in the modulation of synaptic transmission, through which they exert their psychoactive, motor and antinociceptive effects, among others; they also exert extraneural effects, mainly immunomodulation and vasodilation. Recent data suggest that the cannabinoid system might play an important role in human ontogeny and could participate in the implantation and early development of the embryo, in fetal brain development, and in the beginning of breast feeding after birth. In addition, the vasodilatory effect of cannabinoids, together with inhibition of the release of excitotoxic amino acids and cytokines, as well as modulation of oxidative stress and the toxic production of nitric oxide, justify the growing evidence pointing to a possible neuroprotective effect of cannabinoids in perinatal asphyxia. PMID- 16266620 TI - [Non-invasive ventilation in neonates with Ondine syndrome: a real indication?]. AB - Ondine's syndrome is a congenital central hypoventilation syndrome due to a disorder in the autonomic control of breathing in the absence of any primary disease that would explain it. Noninvasive ventilation (NIV) has been reported to be effective in the management of these patients, thus avoiding the need for tracheotomy for prolonged mechanical ventilation. We describe our experience of NIV in infants with Ondine's syndrome. Two infants with Ondine's syndrome were transferred to our center for management and adjustment of therapy. On admission NIV (BiPAP VISION) was started with nasal interphase in S/T (spontaneous/timed) mode, which failed to provide suitable ventilation (PCO2 > 70 mmHg). Finally, tracheotomy for continuous mechanical ventilation was performed. Although NIV has been reported to be successful in some patients with Ondine's syndrome, its application in patients younger than 5 years does not seem to be the general norm of treatment. PMID- 16266621 TI - [Effectiveness of behavioral intervention in nocturnal enuresis]. AB - BACKGROUND: Nocturnal enuresis is a common, often misunderstood condition that affects millions of children. Although the rate of spontaneous resolution is 15 % per year, treatment is often required for social or psychological reasons. OBJECTIVE: To determine the prevalence of nocturnal enuresis in a primary health care area, how was it diagnosed, which treatments were employed and treatment response. MATERIAL AND METHODS: A retrospective study was performed that included all the medical records of children aged between 6 and 14 years old in a basic health area. RESULTS: Twenty-seven children with a diagnosis of nocturnal enuresis were identified (4.3 %). Among these, 21 were boys (77.7 %) and six were girls (22.3 %). There was a family history of nocturnal enuresis in 15 patients (55.5 %). Nocturnal enuresis was not the reason for consultation in 24 of the 27 patients (88.8 %) but was diagnosed collaterally by anamnesis. All the children had simple primary nocturnal (monosymptomatic) enuresis. Five patients abandoned the treatment (18 %), 15 (55.5 %) recovered with behavioral intervention exclusively, the remaining seven children were treated with desmopressin, which was successful in three of them. The remaining four patients were still receiving treatment when the data were collected. CONCLUSIONS: The prevalence of nocturnal enuresis in this primary health area (4.3 %) is similar that reported in other series. In a high percentage (88.8 %) of patients, nocturnal enuresis was not the reason for consultation but was diagnosed during anamnesis. Behavioral intervention was effective in more than half (55.5 %) of the patients. PMID- 16266622 TI - [Risks of black henna tattoos]. AB - Temporary henna tattoos have become increasingly popular in the last few years, because of their apparent harmlessness and disappearance in few weeks. Black henna contains paraphenylenediamine (PPD), a synthetic colorant which is used in hair dyes and can cause sensitization. Many cases of allergic contact dermatitis occurring after tattooing have been reported, especially in children and teenagers during the summer time, since there are many tattoo-painters at holiday places and the parents are unaware of the risks of these tattoos. Clinicians, particularly pediatricians, primary care physicians and dermatologists should seek to remedy this misinformation. We can be the first person to be consulted before the tattoo painting and we will treat patients with lesions. We present our experience of eight cases of allergic contact dermatitis after tattooing and briefly review the literature on the risks of black henna tattoos. PMID- 16266623 TI - [Ingestion of foreign bodies containing lead]. AB - Ingestion of a foreign body containing lead in children carries the additional risk of acute lead poisoning secondary to dissolution and absorption of the ingested lead in the acid environment of the stomach. We report the case of a 3 year-old girl who ingested a lead sinker. The patient was asymptomatic but therapy with a proton pump inhibitor (omeprazole) was empirically started on admission and the foreign body was removed from the stomach by emergency endoscopy within 6 hours of ingestion. Despite the quick removal, blood lead levels (drawn 3 hours after ingestion) exceeded 40 .g/dl. The patient completely recovered and continues to do well after 1 year of follow-up. This case reveals that acute elevations of blood lead concentrations may occur rapidly after ingestion of lead foreign bodies. Even in the absence of symptoms, lead foreign body ingestion in children should prompt lead screening and initiation of appropriate treatment. PMID- 16266624 TI - [Multiple intestinal perforations due to ingestion of magnetized pieces of a toy]. PMID- 16266626 TI - [Atypical presentation of acute paracetamol poisoning]. PMID- 16266625 TI - [Fourteen-year survival in a patient with Edwards syndrome]. PMID- 16266627 TI - [Schonlein-Henoch purpura: a complex case]. PMID- 16266628 TI - [Spectacular response of acute disseminated encephalomyelitis after high-dose steroid therapy]. PMID- 16266629 TI - [Hypoglycemia due to adrenal insufficiency secondary to corticotropin-releasing hormone deficiency]. PMID- 16266630 TI - [Bone infarction as the first manifestation of sickle cell anemia]. PMID- 16266631 TI - [Artificial nutrition in pediatric intensive care units]. PMID- 16266633 TI - [Neonate with a short lower extremity]. PMID- 16266634 TI - [Quality improvement program of nosocomial infection in colorectal cancer surgery]. AB - BACKGROUND AND OBJECTIVE: The surgical-site infection (SSI) is a complication of colorectal neoplasia surgery. The objectives of the study were to identify the SSI risk factors associated with colon surgery and to describe a strategy of quality improvement using surgical-site rates. PATIENTS AND METHOD: Prospective cohort study of in-patients undergoing neoplasia colorectal surgery between 1st July 2002 to 30th June 2003. A descriptive analysis was implemented. Benchmarking was used as tool of quality improvement, and the outcomes were measured using the standardized infection ratio (SIR). To define the risk factors, the Chi square test and logistic regression test were used in univariate and multivariate analysis, respectively. RESULTS: 148 patients were included in the study. The SSI accumulative incidence rate (IA) was 10.14%, and the incidence rate was 6.47 SSI per 1000 days. The SIR was 1.53 the first semester and 1.02 the second one. The multivariate analysis identified two risk factors associated with SSI: unscheduled admission (odds ratio [OR] = 7.47, 95% confidence interval [CI] 2.03 27.48) and a risk index of American Society of Anaesthesiologists (ASA) > or = 3 (OR = 6.77, IC 95%, 1.15-39.84). CONCLUSIONS: An unscheduled admission and high risk ASA index were risk factors associated with SSI in patients undergoing colorectal surgery. The program of quality improvement based on benchmark achieved a reduction of SSI rates similar to the standard ones. PMID- 16266635 TI - [Sociodemographic factors and lifestyle habits associated with weight change in the elderly in Spain]. AB - BACKGROUND AND OBJECTIVE: Our purpose was to examine sociodemographic factors and lifestyle customs associated with weight change in the older adult population of Spain. SUBJECTS AND METHOD: Prospective cohort study conducted from 2001 to 2003 among a cohort of 2,384 people representative of the non-institutionalized Spanish population aged 60 years and over. Data were collected through home interviews, and main statistical analysis was performed through polytomous logistic regression. RESULTS: Over the 2-year follow-up, 27.9% of men and 27.3% of women lost > or = 3 kg, and 18.2% of men and 16.9% of women gained > or = 3 kg. Among men, a loss of > or = 3 kg was more likely among former smokers (odds ratio [OR] = 1.58; 95% confidence interval [CI], 1.13-2.23) and among those with higher body mass index (BMI) (p for linear trend < 0.0001). A weight gain of > or = 3 kg was also more frequent in former smokers (OR = 1.93; 95% IC, 1.28-2.90). In addition, the risk of gaining > or = 3 kg decreased progressively with the increase in the frequency of physical activity at leisure time (p for linear trend < 0.0001). Among women, results were similar to those in men, though in former smokers there was no association with weight changes. CONCLUSIONS: In the older adult population, BMI, physical activity at leisure time, and a former smoker status are associated with weight change in the short term. Intervention on physical activity should be a priority, because it is the main modifiable factor, is associated with health-related quality of life, and it could contribute to avoid excess weight. PMID- 16266636 TI - [Epidemiological surveillance of sexually-transmitted diseases. Spain 1995-2003]. AB - BACKGROUND AND OBJECTIVE: Sexually transmitted infections (STIs) are a major public health problem due to their morbidity, complications and sequels, and also by their relationships with human immunodeficiency virus infection. Several countries have been described a resurgence of STIs since the mid-1990's. MATERIAL AND METHOD: Descriptive study of the STIs monitored by epidemiological surveillance, using as source of information the Spanish National Network of Epidemiological Surveillance data, from 1995 to 2003. RESULTS: In 2003, 1069 new cases of gonorrhoea (rate: 2.55 per 100,000 population) and 917 new cases of syphilis (rate: 2.19 per 100,000 population) were notified to the Spanish Compulsory Disease Reporting System. Between 1995 and 2003, gonorrhoea incidence rates decreased in a 78% and syphilis rates diminish in a 15%. Nevertheless, between 2001 and 2003, both gonorrhoea and syphilis incidence rates have increased in a 29% and 27% respectively. CONCLUSIONS: The data analyzed in this period show an evident change in the trend of the incidence of these STIs. This fact has to be confirmed in the future. PMID- 16266637 TI - [Measurement of blood pressure and uncertainty principle]. PMID- 16266638 TI - [Nutrition assessment in hospitalized patients in Spain]. PMID- 16266639 TI - [Paraneoplasic neurologic syndromes]. PMID- 16266640 TI - [Infectious pulmonary complications in HIV-infected patients in the high by active antiretroviral therapy era in Spain]. AB - Pulmonary complications in HIV-infected patients are at present a first-rate problem. They are the main cause of hospital admission of these patients in our country. Most HIV-patients have a pulmonary complication during the evolution of the infection. The main etiologic diagnosis is bacterial pneumonia, especially pneumococcal pneumonia; the second most frequent cause is Pneumocystis jiroveci (previously named P. carinii) pneumonia and the third cause is mycobacteriosis, particularly Mycobacterium tuberculosis. From early studies, important changes in the epidemiology of HIV-related pulmonary complications have occurred. General prescription of P. jiroveci primary prophylaxis is probably one of the main causes, and, more recently, the use of highly active antiretroviral therapy may also be an underlying explanation. In this review, epidemiology, diagnosis and outcome of HIV-related pulmonary complications in our country are update. PMID- 16266642 TI - [Significant increase of functional status and decrease of fatigue in patients with chronic fatigue syndrome after completing cognitive behavioural group therapy]. PMID- 16266641 TI - [Hallucitations and agitation in two young patients]. PMID- 16266643 TI - [Cost and value of radiation therapy]. PMID- 16266644 TI - [Calcinosis cutis. An inusual complication on adult dermatomyositis]. PMID- 16266645 TI - [Central hypothyroidism related with bexarotene treatment]. PMID- 16266647 TI - [Health promotion, community care, and family medicine]. PMID- 16266648 TI - [Perceptions and expectations on primary health care: a new form of identifying improvements in the care system]. AB - OBJECTIVE: To analyse the opinions of the users of primary care on the care that they receive and to identify the principal areas of satisfaction. DESIGN: Qualitative study using discussion groups and open interviews during the period January-May 2003. SETTING: Health areas of Valladolid, Spain. PARTICIPANTS: The inclusion criteria were: to have attended a primary care clinic at sometime and to be between 35 and 80 years old. Recruitment was carried out through key informants, using the snowball technique. METHOD: 6 discussion groups and interviews with representatives of 3 nursing and 1 residents association were carried out. Structural sampling was carried out as regards the variables that influenced satisfaction. The conversations were recorded using tape recorders and literally transcribed on paper. The analysis of the texts was carried out by 2 investigators and concordance was sought between them. RESULTS: The principal areas related to satisfaction were: the treatment received from the professionals, which is considered a fundamental part of care, combined with the technical quality, continuity of the care, the admission services, the bureaucratic procedures, the barriers for accessing specialised services, and waiting lists. CONCLUSIONS: Personalized care, the time dedicated by the professional, the continuity of care, and waiting lists are the principal areas related to the perceived satisfaction of the patients. The possible responses to improve this situation are: the implementation of changes in the care management and organisation which would simplify the procedures, investment of resources (human and economic), changes in the model of the professional-patient relationship, and improvements in undergraduate and postgraduate training. PMID- 16266649 TI - [On the perceived quality of primary care]. PMID- 16266650 TI - [Efficacy of educational sessions to modify the prescription of new drugs]. AB - OBJECTIVE: To evaluate the efficacy of an educational intervention to minimise the prescription of those new medicines whose therapeutic effects are of little benefit. DESIGN: Controlled and randomised experimental study. SETTING: 27 health centres in the province of Sevilla, Spain. PARTICIPANTS: 376 general practitioners. The 264 who worked in the same posts were randomised for the 6 pre intervention months. 10 of them did not complete the post-intervention period. INTERVENTIONS: Four 45-minute training sessions in a 2-month period, given by health team doctors, with a critical reading of the studies available on recently marketed drugs, plus personal feed-back on prescription and bulletins on therapeutic novelties. The control group received only the feed-back and bulletins. MAIN MEASUREMENTS: Prescription of new medication of little benefit, measured as the number of packages out of the total. Second, the amount of coxib and eprosartan measured as defined daily doses. RESULTS: In the 6 months after the educational sessions, the doctors in the intervention group prescribed proportionately fewer therapeutic novelties of little benefit than those allocated to the control group (1.34% vs 1.62%; P<.001). The coxib and eprosartan prescribed showed only a non-significant trend towards less prescription by the intervention group. CONCLUSIONS: The group educational sessions, run by doctors trained in aspects of evidence-based medicine and prepared jointly with the pharmacy unit, reduced discreetly the prescription of new medicines that were not very innovative. PMID- 16266651 TI - [Costs of spirometry as a screening test for chronic obstructive pulmonary disease in primary care]. AB - OBJECTIVE: To calculate the costs of COPD screening by simple spirometry, conducted in primary care, in smokers and ex-smokers over 40 years old. DESIGN: Descriptive study. Cost analysis. SETTING: Semi-rural health district covering some 6000 inhabitants. PARTICIPANTS: Randomised sample of 350 smokers and ex smokers over 40 allocated to the centre. 73 were excluded on SEFAR criteria. INTERVENTIONS AND MEASUREMENTS: Simple spirometry was conducted with an automatic, portable, dry spirometer. Costs were calculated from the time needed, the material, human resources and the number of valid spirometer readings. RESULTS: 85% of 277 had a spirometry test. 18% of the spirometry readings were not valid. 3 spirometries an hour could be done: at least 1 was pathological and a bronchodilator test was needed. The cost of a spirometry, if a professional devotes 2 hours a day for 5 years, was 10.57 euros or 8.54 euros, for doctor or nurse, respectively. Modifying the number of spirometries per year or the number of professionals trained for such a technique causes only minor changes in cost. CONCLUSIONS: Optimal cost: 9000 spirometries a year by a single nurse. However, this would entail one nurse's almost exclusive dedication. Two trained professionals raises the cost slightly and distributes the work load better, enabling more hours to be covered. Before advising any COPD screening, its costs and its real possibility of affecting active smokers (the sole preventive possibility) should be carefully assessed. PMID- 16266652 TI - [Analysis of the new active principles sold in Spain (1992-2002)]. AB - OBJECTIVES: To describe the active principles (AP) marketed in Spain from 1992 to 2002, to determine their characteristics, and to find whether they supposed genuine therapeutic advances (TA). DESIGN: Transversal, descriptive study. MAIN MEASUREMENTS: The degree of TA in the AP analysed was studied with the classification used by the FDA (A*: exceptional therapeutic novelty; A: important therapeutic novelty; B: modest therapeutic improvement; C: null or very little therapeutic improvement, corresponding to "me-too" drugs; and D: not classified), the context of use and the price. RESULTS: 369 new AP were marketed. 3.5% were in group A*, 11.9% in A, 30.1% in B, 49.3% in C, and 5.1% in D. 42.3% corresponded to AP used in hospitals for therapy or diagnosis. Significant differences were found (P<.05) on comparing the degree of TA and the context of use, such that more AP in the A/A* (32.6%) and B (44.0%) groups were found in AP used in hospital therapy and diagnosis than in AP used in primary care and generally (5.3% in the A/A* groups and 23.4% in group B). Only 11 AP of the A/A* groups were used in primary care. The cost per defined daily dose was 17.6 euros; and the new AP in group C were dearer than already existing alternatives in 93.4% of cases. CONCLUSIONS: Real TA are few in number and preferentially used in hospitals. Almost all the new AP are "me-too" drugs and are dearer than already existing alternatives. PMID- 16266653 TI - [Psychiatric/social guidance and family care in family medicine: bad times for lyrical poetry]. PMID- 16266654 TI - [Health utility of unfitness for work]. PMID- 16266655 TI - [Flu vaccination in primary care: analysis of the process and proposals for increasing coverage]. AB - INTRODUCTION: Flu vaccination (FV) in elderly patients at risk is linked to a 50% 60% reduction in hospital admissions and up to an 80% drop in deaths from complications arising from the illness. Equally clear benefits have been found for other risk groups, such as patients with chronic cardiovascular or respiratory diseases. The vaccine is cost-effective for both the elderly and other risk groups. Despite this, vaccination rates are low, even among health staff. OBJECTIVES: To update our knowledge of FV by means of a review of the bibliography and to describe a series of interventions that have proved successful in increasing vaccination rates. PROGRAMME: To discover procedures, the following factors were analysed: the environment, patients and health professionals that condition vaccination, the characteristics of the health-care organisation for the vaccination campaign, and the clinical organisation of risk groups required. DISCUSSION: After this analysis, certain communicated strategies that manage to increase vaccination coverage and others that could be introduced into primary care were discussed. We conclude that, given the clinical evidence available and the ease of introducing certain other interventions, improvement of flu vaccination procedures and increased vaccine coverage of patients at risk is not only advisable, but is an ethical imperative. Improvements that are within the possibilities of every primary care clinic could be introduced. PMID- 16266656 TI - [Care for immigrant minors in the consultation: between cultural difference and crime. A case of clitoral ablation]. PMID- 16266657 TI - [The Constitution of the Spanish Association of Teaching Committees (AREDA)]. PMID- 16266658 TI - [Tendinitis linked to taking norfloxacin: concerning a case]. PMID- 16266659 TI - [Descriptive study of adaptive disorders in a primary care centre]. PMID- 16266660 TI - [Half the capillary glycaemia strips that our patients use have doubtful indication]. PMID- 16266661 TI - [Indications authorized in Spain of osteoporosis drugs]. PMID- 16266662 TI - [The last will be first]. PMID- 16266663 TI - [Calculation of cardiovascular risk on the Framingham scale calibrated by REGICOR]. PMID- 16266664 TI - [The lung at high altitudes]. PMID- 16266665 TI - [Analysis of exhaled breath condensate: a technique with a future?]. PMID- 16266666 TI - [Limitations of the technique to determine hydrogen peroxide levels in exhaled breath condensate from patients with adult respiratory distress syndrome]. AB - OBJECTIVE: Exhaled breath condensate represents an alternative to bronchoalveolar lavage for the analysis of markers of inflammation and oxidative stress in patients with adult respiratory distress syndrome (ARDS). However, analysis of hydrogen peroxide (H2O2) yields variable results that do not correlate with severity of the clinical presentation. In an attempt to explain this variability, the aim of the present study was to assess the possible limitations of the most commonly used technique for analyzing H2O2 in breath condensate. PATIENTS AND METHODS: H2O2 levels were analyzed using the Gallati technique (linear range between 0.3 and 10 microM, r=0.99; P<.05) in serial samples of condensate taken from the expiratory tube of a mechanical ventilator in 6 patients with ARDS. RESULTS: The volume of condensate obtained correlated to minute ventilation (r=0.96; P<.05). In 11 out of 23 samples, a spectrophotometer reading was obtained at 450 nm despite the absence of the characteristic color of the reaction and in some of these samples a spontaneous reading was obtained that was indicative of contamination. The absorbance spectrum of these samples did not contain the characteristic peak for H2O2 at 450 nm and pretreatment of some samples with catalase did not affect the absorbance at 450 nm. CONCLUSIONS: The spectrophotometric method commonly used to measure H2O2 levels in breath condensate lacks specificity in ARDS due to the presence of variable levels of contaminants in the samples, which lead to false positives. PMID- 16266667 TI - [Changes in spirometric parameters and arterial oxygen saturation during a mountain ascent to over 3000 meters]. AB - OBJECTIVE: To ascertain whether climbing a mountain over 3000 meters high produces any alterations in ventilation, whether such alterations are modified by acclimatization, and whether they correlate with changes in arterial oxygen saturation (SaO2) or the development of acute mountain sickness (AMS). SUBJECTS AND METHODS: The following parameters were measured in 8 unacclimatized mountaineers who climbed Aneto (3404 m) and spent 3 days at the summit: forced vital capacity (FVC), forced expiratory volume in 1 second (FEV1), airway response to inhaled terbutaline, SaO2, and the symptoms of AMS. RESULTS: At the summit, mean (SD) FEV1 declined by 12.3% (5.7%) and mean FVC by 7.6% (6.7%) while the ratio of FEV1 to FVC remained normal. The means for both parameters were higher on the following day. No airway response to bronchodilator treatment was observed. The restriction disappeared entirely on descent. At the peak, SaO2 increased progressively as the climbers became acclimatized. During the ascent, FEV1 correlated with SaO2 (r=0.79). One participant who suffered from AMS had a ratio of FEV1 to FVC less than 70% and the worst SaO2 during the 3 days on the summit. Obstruction preceded the AMS symptoms, did not respond to bronchodilator treatment, and disappeared when the climber descended. CONCLUSIONS: The mountaineers who climbed over 3000 meters presented restriction that correlated with hypoxemia. This restriction did not respond to bronchodilator treatment, improved with acclimatization, and disappeared on descent. One person with AMS presented obstruction that did not respond to terbutaline and disappeared on descent. PMID- 16266668 TI - [Cystic fibrosis in adults: inter- and intraobserver agreement for the Brasfield and Chrispin-Norman chest radiography scoring systems and correlation with clinical data and spirometry]. AB - OBJECTIVE: Most chest radiography scoring systems for patients with cystic fibrosis have been developed for children but are also used for adults. Our aim was to evaluate the intra- and interobserver variability of 2 radiographic scoring systems in adults with cystic fibrosis and to assess the correlation of these systems with clinical and spirometric parameters. PATIENTS AND METHODS: The chest x-rays of 24 adult patients with cystic fibrosis were compared using 2 scoring systems (Brasfield and Chrispin-Norman). The x-rays were scored by 2 radiologists and reevaluated 4 months later by 1 of the 2 observers. Intra- and interobserver agreement was assessed using the intraclass and Pearson's correlation coefficients. The radiographic scores were compared to lung function tests and other clinical data. RESULTS: Both intra- and interobserver agreement were high (r > or = 0.9 and the intraclass correlation coefficient > or = 0.85 with both systems for both samples). Both scoring systems correlated with spirometry results: forced expiratory volume in the first second (FEV1) (r = 0.64 and r = 0.55), FEV1% (r = 0.75 and r = 0.72), and the percentage of forced vital capacity in relation to the predicted value (r = 0.63 and r = 0.056). We found no association between scoring system and sex, age, or body mass index. CONCLUSIONS: Assessment of chest radiographs of adult patients with cystic fibrosis by the Brasfield and Chrispin-Norman scoring systems shows good intra- and interobserver agreement. Both systems correlate well with lung function variables, especially FEV1. PMID- 16266669 TI - [Nontuberculous mycobacteria in patients with cystic fibrosis]. AB - OBJECTIVE: Patients with cystic fibrosis are at great risk of infection by nontuberculous mycobacteria from the environment because of certain predisposing factors such as bronchiectasis, malnutrition, and diabetes. The aim of this study was to analyze the mycobacterial content of sputum smears and cultures from adult patients with cystic fibrosis attended at a specialized unit for adults from March 1997 through December 2001. PATIENTS AND METHODS: Sputum samples were collected prospectively according to a protocol applied at each visit, and during most exacerbations staining and culture for mycobacteria were ordered in addition to the usual cultures for bacteria and fungi. A tuberculin test was performed at the end of the study. RESULTS: Twenty-eight patients (16 men) with cystic fibrosis were enrolled. The mean (SD) age was 25.3 (6.7) years. A total of 251 samples were cultured (range in number of samples per patient, 1-31). The mean period of follow up was 40.3 (22.1) months. The sputum smear was positive in 29 cases (4 patients); the culture was positive in 7 patients. More than 3 samples were positive in only 4 patients. Mycobacterium abscessus was isolated in 3 cases, Mycobacterium avium complex in 2 and Mycobacterium simiae in 1 and other an unidentified rapid growth Mycobacterium species. The Mantoux test was positive in 5 patients. Two of the 4 patients in whose samples mycobacteria were isolated repeatedly required treatment. CONCLUSIONS: The prevalence of nontuberculous mycobacterial infection is high in patients with cystic fibrosis. Staining and culture for mycobacteria should be carried out regularly and whenever exacerbation of pulmonary symptoms cannot be attributed to bacteria usually found in such patients. Patients with recurrent isolations of mycobacteria should be monitored closely. PMID- 16266670 TI - [Variability among pathologists in the histological diagnosis of diffuse interstitial lung diseases]. AB - OBJECTIVE: Diffuse interstitial lung diseases (DILD) form a group of diseases which affect the alveolar interstitial space and share very similar clinical, radiological, and functional features, making lung biopsy essential for establishing diagnosis, prognosis, and treatment in many cases. We aimed to see whether there was agreement in histopathological diagnosis among different groups of pathologists in their assessment of these diseases. MATERIAL AND METHODS: Biopsies were studied from 33 patients suffering from noninfectious, nontumorous DILD. The biopsies had been assessed by 2 groups of pathologists: one specializing in this type of disease and another which was not a specialist group. RESULTS: There was disagreement in the histology reports of 10 out of the 33 cases studied (30.3%): 9 cases in the group of 22 cases of idiopathic interstitial pneumonia (40.9%) and 1 in the group of 3 DILD with known or associated causes. No discrepancies were found, however, in the diagnosis of primary DILD or DILD associated with other, less well-defined processes. CONCLUSIONS: We believe that idiopathic interstitial pneumonias are the DILD which pose most problems for pathologists. Therefore, the study of DILD requires specific dedication by pathologists and other professionals and specialists. PMID- 16266671 TI - [Rhinitis and asthma: one airway, one disease]. PMID- 16266672 TI - [Setting up and organizing a noninvasive ventilation unit for hospital and home therapy]. PMID- 16266673 TI - [Exhaled breath condensate: standardized collection of samples from healthy volunteers]. AB - Expired breath condensate collection is a noninvasive technique for obtaining a sample in which to analyze substances that reflect the functional status of the lung and other tissues. Twenty healthy volunteers provided 3 expired breath samples: the second was collected 20 minutes after the first and the third 48 hours after the first. The air and condensate volumes were assessed. The mean (SD) volume of condensate in exhaled air over a period of 15 minutes was 1.8 (0.5) mL (95% confidence interval [CI], 1.5-2 mL) and the coefficient of variation was 29%. Analysis of variance in the 3 samples demonstrated no significant differences. The mean volume of air inhaled over 15 minutes was 119 (25) L (95% CI, 112-125 L). These results indicate that it takes at least 15 minutes and the inhalation of some 120 L of air to collect a condensate volume that exceeds 1.5 mL, sufficient to allow distribution in aliquots to analyze fundamental physical and chemical properties (conductivity, pH) and certain relevant biomarkers. PMID- 16266674 TI - [Diffuse pulmonary calcification in a patient with renal insufficiency]. AB - We report the case of a 37-year-old man with chronic renal insufficiency, on hemodialysis, with no respiratory symptoms but whose chest radiograph showed parenchymal consolidation in the middle and upper lung fields. High resolution computed tomography showed a high-attenuating diffuse alveolar pattern that indicated calcium deposits. Bronchoscopy revealed metastatic calcification on the interalveolar septa and bronchiolar and arteriolar. The present report, based on radiologic and bronchoscopic findings, describes the pathogenesis and anatomical distribution of the patient's diffuse pulmonary calcification. PMID- 16266675 TI - [Tuberculosis as an occupational disease]. PMID- 16266676 TI - Neurotropism and neuropathological effects of selected rhabdoviruses on intranasally-infected newborn mice. AB - Viral neurotropism is the ability of viruses to infect neuronal cells. This is well studied for herpesviruses, rabies-related viruses, and a few others, but it is poorly investigated among almost all arboviruses. In this study, we describe both the neurotropism and the neuropathological effects of Amazonian rhabdoviruses on the brains of experimentally infected-newborn mice. Suckling mice were intranasally infected with 10(-4) to 10(-8) LD50 of viruses. Animals were anaesthetized and perfused after they had become sick. Immunohistochemistry using specific anti-virus and anti-active caspase three antibodies was performed. All infected animals developed fatal encephalitis. Survival time ranged from 18 h to 15 days. Viruses presented distinct species-dependent neurotropism for CNS regions. Histopathological analysis revealed variable degrees of necrosis and apoptosis in different brain regions. These results showed that viruses belonging to the Rhabdoviridae family possess distinct tropism for CNS structures and induce different pattern of cell death depending on the CNS region. PMID- 16266677 TI - Amperometric, screen-printed, glucose biosensor for analysis of human plasma samples using a biocomposite water-based carbon ink incorporating glucose oxidase. AB - This paper describes the optimisation of a screen-printing water-based carbon ink containing cobalt phthalocyanine (CoPC) and glucose oxidase (GOD) for the fabrication of a glucose biosensor. To optimise the performance of the biosensor, the loadings of the electrocatalyst (CoPC) and enzyme (GOD) were varied. It was found that the maximum linear range was achieved with a CoPC loading of 20% (m/m, relative to the mass of carbon) and a GOD loading of 628 U per gram of carbon. In our studies we chose to employ chronoamperometry, as this technique is commonly used for commercial devices. The optimum operating applied potential was found to be +0.5 V, following an incubation period of 60 s. The optimum supporting electrolyte was found to be 0.05 M phosphate buffer at pH 8.0, which resulted in a linear range of 0.2-5 mM, the former represents the detection limit. The sensitivity was 1.12 microA mM(-1). The effect of temperature was also investigated, and it was found that 40 degrees C gave optimal performance. The resulting amperometric biosensors were evaluated by measuring the glucose concentrations for 10 different human plasma samples containing endogenous glucose and also added glucose. The same samples were analysed by a standard spectrophotometric method, and the results obtained by the two different methods were compared. A good correlation coefficient (R(2) = 0.95) and slope (0.98) were calculated from the experimental data, indicating that the new devices hold promise for biomedical studies. PMID- 16266678 TI - A homogeneous, high-throughput fluorescence resonance energy transfer-based DNA polymerase assay. AB - A homogeneous, fluorescence resonance energy transfer (FRET)-based DNA polymerase assay that is suitable for high-throughput screening for inhibitors, and can also be used for steady-state kinetic investigations, is described. The activity, kinetic mechanism, and processivity of the isolated alpha subunit of DNA polymerase III, the product of the dnaE gene, from the gram-negative pathogen Haemophilus influenzae were investigated using the FRET assay. PMID- 16266679 TI - The analysis of folate and its metabolic precursors in biological samples. PMID- 16266680 TI - A continuous spectrophotometric assay method for peptidylarginine deiminase type 4 activity. AB - A simple, continuous spectrophotometric assay for peptidylarginine deiminase (PAD) is described. Deimination of peptidylarginine results in the formation of peptidylcitrulline and ammonia. The ammonia released during peptidylarginine hydrolysis is coupled to the glutamate-dehydrogenase-catalyzed reductive amination of alpha-ketoglutarate to glutamate and reduced nicotinamide adenine dinucleotide (NADH) oxidation. The disappearance of absorbance at 340nm due to NADH oxidation is continuously measured. The specific activity obtained by this new protocol for highly purified human PAD is comparable to that obtained by a commonly used colorimetric procedure, which measures the ureido group of peptidylcitrulline by coupling with diacetyl monoxime. The present continuous spectrophotometric method is highly sensitive and accurate and is thus suitable for enzyme kinetic analysis of PAD. The Ca(2+) concentration for half-maximal activity of PAD obtained by this method is comparable to that previously obtained by the colorimetric procedure. PMID- 16266681 TI - A polypyrrole protein microarray for antibody-antigen interaction studies using a label-free detection process. AB - Protein microarray is a promising technology that should combine rapidity and easy use with high throughput and versatility. This article describes a method in which an electrocopolymerization process is employed to graft biological molecules on to a chip so that surface plasmon resonance imaging may be used to detect molecular interactions. Copolymerization of pyrrole-modified protein and pyrrole is an efficient grafting process which immobilizes molecules at defined positions on a gold surface. Surface plasmon resonance imaging is an optical technique that allows real-time simultaneous detection of molecular interactions on a large number of spots without labeling. This method was successfully used to analyze antibody-antigen interactions. This illustrates its high specificity and good sensitivity and demonstrates its suitability for biological studies. PMID- 16266682 TI - New immobilization method for immunoaffinity biosensors by using thiolated proteins. AB - A new immobilization method for immunoaffinity (IA) biosensors that ensures the high surface density and the stability of the IA layer was developed. For the immobilization of biomolecules, the molecular recognition protein was first thiolated by covalent conjugation of mercaptopropionic acid, and then the thiolated protein was attached on the gold surface of the transducer. In this work, horseradish peroxidase (HRP) and its antibody were used as a model antigen antibody, and the following properties of the IA layer prepared by thiolated protein were estimated: (i) biological integrity of HRP after the immobilization process by using activity assay, (ii) charge transfer resistance by immobilization, (iii) mass loading by the surface plasmon resonance (SPR) biosensor, (iv) number of binding sites, and (v) feasibility test for the measurement of capacitive change by the antigen-antibody interaction. Based on these parameters, the immobilization method by using thiolated protein was determined to be feasible for application to IA biosensors. PMID- 16266683 TI - Determination of saccharide content in pneumococcal polysaccharides and conjugate vaccines by GC-MSD. AB - A simple and sensitive gas chromatographic method was designed for quantitative analysis of Streptococcus pneumoniae capsular polysaccharides, activated polysaccharides, and polysaccharide conjugates. Pneumococcal serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, and 23F polysaccharide or conjugate were subjected to methanolysis in 3N hydrochloric acid in methanol followed by re-N acetylation and trimethylsilylation. Derivatized samples were chromatographed and detected using gas chromatography with mass selective detector. Gas chromatographic results were compared with colorimetric values with agreement of 92 to 123% over the range of all samples tested. Monosaccharides released during methanolysis included hexoses, uronic acids, 6-deoxy-hexoses, amino sugars, and alditols. Quantitative recovery of monosaccharides was achieved for all serotypes by the use of a single methanolysis, derivatization, and chromatography procedure. Response factors generated from authentic monosaccharide standards were used for quantitation of pneumococcal polysaccharides and conjugates with confirmation of peak assignments by retention time and mass spectral analysis. This method allows saccharide quantitation in multivalent pneumococcal vaccine intermediates and final drug products with low-level detection (10 pg) and peak purity. PMID- 16266684 TI - Potentiometric sensor for the measurement of Cd2+ transport in yeast and plants. AB - Research on heavy metals, and especially on transport of Cd(2+), has attracted much interest during the past decade. An optimized Cd(2+)-selective electrode for the continuous potentiometric monitoring of Cd(2+) fluxes in biological systems is presented. The selectivity of the electrode for Cd(2+) was further improved, and it now has very good long-term stability. The utility of this simple and inexpensive method is demonstrated by studying the Cd(2+) transport with model organisms, such as the yeast Saccharomyces cerevisiae and Arabidopsis cell cultures, frequently used in plant science. Its lower detection limits in the presence of commonly used growth media for yeast and plant cells are improved by approximately three orders of magnitude and are 10(-10) and 10(-8)M Cd(2+), respectively. Control experiments using atomic absorption spectrophotometry confirm that the decrease in Cd(2+) activities in the cell cultures is indeed due to the uptake of these metal ions by the cells. Both model systems can be easily transformed; therefore, in combination with the new electrode, they are very promising tools for the investigation of any protein of interest that might be involved in Cd(2+) transport. PMID- 16266685 TI - Small-scale total DNA extraction from bacteria and yeast for PCR applications. PMID- 16266686 TI - Optimization of fragmentation conditions for microarray analysis of viral RNA. AB - An important consideration in microarray analysis of nucleic acids is the efficiency with which the target molecule is captured by, or hybridized to, surface-immobilized oligos. For RNA, secondary and tertiary structure of the target strand can significantly decrease capture efficiency. To overcome this limitation, RNA is often fragmented to reduce structural effects. In this study, the metal ion-catalyzed base hydrolysis fragmentation conditions for viral RNA extracted from influenza viruses were evaluated and the hybridization efficiency of the resulting fragments was determined as a function of fragment length. The amount of RNA captured was evaluated qualitatively by fluorescence intensity normalized to an internal standard. Optimized conditions for influenza RNA were determined to include a fragmentation time of 20-30 min at 75 degrees C. These conditions resulted in a maximum concentration of fragments between 38 and 150 nt in length and a maximum in the capture and label efficiency. PMID- 16266687 TI - Probing the role of the hyper-reactive histidine residue of arginase. AB - Rat liver arginase (arginase I) is potently inactivated by diethyl pyrocarbonate, with a second-order rate constant of 113M(-1)s(-1) for the inactivation process at pH 7.0, 25 degrees C. Partial protection from inactivation is provided by the product of the reaction, l-ornithine, while nearly complete protection is afforded by the inhibitor pair, l-ornithine and borate. The role of H141 has been probed by mutagenesis, chemical modulation, and X-ray diffraction. The hyper reactivity of H141 towards diethyl pyrocarbonate can be explained by its proximity to E277. A proton shuttling role for H141 is supported by its conformational mobility observed among the known arginase structures. H141 is proposed to serve as an acid/base catalyst, deprotonating the metal-bridging water molecule to generate the metal-bridging hydroxide nucleophile, and by protonating the amino group of the product to facilitate its departure. PMID- 16266688 TI - Delta (delta) opioid receptors in small and medium-sized trigeminal neurons supporting the dental pulp of rats. AB - The control of pain perception is a challenge in clinical dentistry, most prominent during tooth pulp inflammation. The tooth pulp is a well-defined target, and is densely supplied by a sensory trigeminal innervation. Opioids are signaling molecules that are suggested to participate in pain perception. Here we analysed the presence of delta opioid receptor (DOR) in trigeminal neurons innervating the tooth pulp of rat molars. Immunohistochemical and ultrastructural analysis revealed that DOR was identified in peripheral nerves in the molar dental pulp, both in the root and the coronal pulpal parts, with branching in the highly innervated subodontoblast layer. DOR was localised in about one third of all the trigeminal dental neurons, identified by means of retrograde neuronal transport of fluorogold (FG) from the dental pulp. Of the DOR-labeled neurons, nearly all were small and medium-sized (147.5-1,810.2 microm(2), mean 749.1 +/- 327.3 microm(2)). Confocal microscopy confirmed that DOR-immunoreactivity was distributed as granules in the neuronal cytoplasm. Approximately 70% of the DOR immunoreactive neurons were also immunopositive for vanilloid receptor 1 (TRPV1). Ultrastructural analysis demonstrated DOR-immunoreactivity in the unmyelinated and in some of the myelinated nerve fibers in the dental pulp. These results indicate that DOR may influence the function in a subset of small and medium sized trigeminal sensory neurons supporting the tooth, which are mainly known for their ability to mediate nociceptive stimuli. Agonists, acting on DOR, may thus have an influence on a subpopulation of nociceptive neurons supporting the rat tooth. PMID- 16266689 TI - Different glycoforms of the human GPI-anchored antigen CD52 associate differently with lipid microdomains in leukocytes and sperm membranes. AB - CD52 is a human GPI-anchored antigen, expressed exclusively in the immune system and part of the reproductive system (epididymal cells). Sperm cells acquire the antigen from the epididymal secretions when transiting in the epididymal corpus and cauda. The peptide backbone of CD52, consisting of only 12 aminoacids, is generally considered no more than a scaffold for post-translational modifications, such as GPI-anchor and especially N-glycosylation which occur at the third asparagine. The latter modification is highly heterogeneous, especially in the reproductive system, giving rise to many different glycoforms, some of which are tissue specific. A peculiar O-glycan-containing glycoform is also found in reproductive and immune systems. We determined to locate CD52 in microdomains of leukocytes and sperm membranes using two antibodies: (1) CAMPATH-1G, the epitope of which includes the last three aminoacids and part of the GPI-anchor of glycoforms present in leukocytes and sperm cells; (2) anti-gp20, the epitope of which belongs to the unique O-glycan-bearing glycoform also present in both cell types. Using a Brij 98 solubilization protocol and sucrose gradient partition we demonstrated that the CD52 glycoforms recognized by both antibodies are markers of typical raft microdomains in leukocytes, whereas in capacitated sperm the O glycoform is included in GM3-rich microdomains different from the cholesterol and GM1-rich lipid rafts with which CAMPATH antigen is stably associated. The importance of the association between GM3 and O-glycans for formation of specialized microdomains was confirmed by heterologous CD52 insertion experiments. When prostasomes from human seminal fluid were incubated with rat sperm from different epididymal regions, the CD52 glycoform recognized by anti gp20 decorated rat epididymal corpus and cauda sperm, associated with the same low-cholesterol GM3-rich sperm membrane fractions as in human sperm. The glycoforms recognized by CAMPATH-1G were not found in rat sperm. The relationship between this differential insertion and differences in glycosylation of rat and human CD52 is discussed. PMID- 16266690 TI - Benzodiazepine involvement in LTP of the GABA-ergic IPSC in rat hippocampal CA1 neurons. AB - Benzodiazepine binding sites are present on gamma-aminobutyric acid (GABA) receptors in hippocampal neurons. Diazepam is known to potentiate the amplitude and prolong the decay of GABA(A) receptor-mediated inhibitory postsynaptic currents (IPSCs). In this study, benzodiazepine involvement in long-term potentiation (LTP) of the IPSC was examined. Whole-cell recordings of IPSCs were made from rat hippocampal CA1 neurons in a slice preparation. LTP was induced by a tetanic stimulation in the stratum radiatum (2 trains of 100 Hz for 1 s, 20 s inter-train interval) while pharmacologically blocking ionotropic glutamate receptors. During LTP, the amplitude of the IPSCs was potentiated in the majority of neurons with the IPSC decay and shape unaffected. Diazepam (5 microM) potentiated the IPSC amplitude and prolonged the decay when applied before, but not during, LTP. In neurons in which LTP could not be induced by a tetanic stimulation, diazepam did not increase the amplitude of the pre-tetanic IPSC. Flumazenil, at a concentration (10 microM) that blocked the enhancement of the IPSC by applied diazepam, had no effect on the IPSC amplitude when applied before LTP induction but significantly decreased the IPSC when applied during LTP maintenance. The antagonist, when applied during the tetanic stimulation, did not block LTP, suggesting that benzodiazepine receptors do not participate in LTP induction. These results indicate that the maintenance of LTP of the IPSC involves (a) the release of endogenous benzodiazepine agonist(s) and/or (b) the participation of benzodiazepine binding sites on subsynaptic GABA(A) receptors. PMID- 16266691 TI - Local inhibition of organic cation transporters increases extracellular serotonin in the medial hypothalamus. AB - In the rat dorsomedial hypothalamus (DMH), serotonin (5-HT) concentrations are altered rapidly in response to acute stressors. The mechanism for rapid changes in 5-HT concentrations in the DMH is not clear. We hypothesize that the mechanism involves corticosteroid-induced alterations in the uptake of 5-HT from extracellular fluid through the action of corticosterone-sensitive organic cation transporters (OCTs). To determine if OCTs affect the clearance of 5-HT from the extracellular fluid compartment within the medial hypothalamus (MH), the OCT blocker, decynium 22 (0, 10, 30, or 100 microM), was perfused into the MH via a microdialysis probe, and dialysate 5-HT concentrations were measured at 20 min intervals. In addition, home cage behavior was measured both before and after drug administration. Inhibition of OCTs in the MH resulted in a reversible dose dependent increase in extracellular 5-HT concentration. Increases in extracellular 5-HT concentrations were associated with increases in grooming behavior in rats treated with the highest concentration of decynium 22. No other behavioral responses were observed following administration of any concentration of decynium 22. These findings are consistent with the hypothesis that OCTs in the MH play an important role in the regulation of serotonergic neurotransmission and specific behavioral responses. Because the MH plays an important role in the neuroendocrine, autonomic, and behavioral responses to stress-related stimuli, these data lead to new questions regarding the role of interactions between corticosterone and corticosterone-sensitive OCTs in stress-induced 5-HT accumulation within the MH as well as the physiological and behavioral consequences of these interactions. PMID- 16266692 TI - Preservation of pressure-induced cutaneous vasodilation by limiting oxidative stress in short-term diabetic mice. AB - OBJECTIVE: Pressure-induced vasodilation (PIV) allows skin blood flow to increase in response to locally applied pressure and may be protective against pressure ulcers. We previously showed that PIV was absent in 1-week diabetic mice exhibiting no neuropathy. Our aim was to determine whether the diabetes-induced PIV alteration could be prevented. METHODS AND RESULTS: Diabetic mice received no treatment or a daily treatment with either sorbinil, alagebrium or alpha-lipoic acid (LPA) for 1 week. Laser Doppler flowmetry was used to evaluate PIV as well as endothelium-dependent vasodilation following iontophoretic delivery of acetylcholine (ACh). The effect of each treatment on oxidative stress was examined by plasma 8-isoprostane assay. LPA was the sole treatment to prevent both PIV and ACh vasodilation alterations, with a significant reduction of oxidative stress in diabetic mice. Both PIV and ACh-vasodilation were abolished in LPA-treated diabetic mice following injection of Nomega-nitro-L-arginine (p<0.05). In contrast, alagebrium and sorbinil prevented neither diabetes-induced PIV abolition nor endothelial alteration. CONCLUSIONS: LPA treatment significantly reduced the oxidative stress and was able to preserve endothelial nitric oxide availability in the cutaneous microcirculation and then to preserve the PIV response in diabetic mice. LPA treatment could play a key role in limiting the risk of pressure-induced cutaneous ulcer during diabetes. PMID- 16266693 TI - Matrix metalloproteinases regulate migration, proliferation, and death of vascular smooth muscle cells by degrading matrix and non-matrix substrates. AB - Intimal thickening occurs in blood vessels in response to injury or atherosclerosis. The balance of migration and proliferation of vascular smooth muscle cells (VSMC) over death by apoptosis has an important impact on the final size of intimal thickening and may also affect atherosclerotic plaque stability. All aspects of VSMC behaviour are under coordinated control by growth factors, cell-matrix and cell-cell interactions. We review the evidence that matrix degrading metalloproteinases (MMPs) regulate migration, proliferation and survival of VSMC. Moreover, we discuss critically the underlying mechanisms, which include changing growth factor availability and remodelling cell-matrix and cell-cell contacts. We conclude that MMPs influence VSMC behaviour by cleaving both matrix and non-matrix substrates. PMID- 16266694 TI - Novel isosorbide-based substrates for human butyrylcholinesterase. AB - Butyrylcholinesterase [EC 3.1.1.8] present widely in mammalian tissue does not have a precisely defined biological function or known endogenous substrate. However, it plays an important role in the detoxification of certain xenobiotics and is an established vector for the systemic liberation of other drugs from their prodrugs. While investigating a series of isosorbide-based prodrugs, we discovered that BuChE catalyses the hydrolysis of esters of the simple sugar isosorbide with unusually rapidity and in some cases with remarkable regioselectivity. In this study, a series of isosorbide esters were synthesised and their rates of hydrolysis measured by HPLC following incubation in diluted plasma solution. In general, little hydrolysis of the 5-ester group could be observed but the 2-ester group was usually hydrolysed very rapidly and the hydrolysis rate exhibited an unusual dependence on the identity of the 5-group. The results indicate that while the 5-ester group is not itself hydrolysed it is important for productive binding in isosorbide diesters. PMID- 16266695 TI - Estimation of oxime efficacy in nerve agent poisoning: a kinetic approach. AB - Standard treatment of poisoning by organophosphorus compounds (OP) includes the administration of an anti-muscarinic, e.g. atropine, and of an acetylcholinesterase (AChE) reactivator (oxime). Two oximes, obidoxime and pralidoxime (2-PAM), are presently commercially available, yet, these compounds are considered to be of insufficient efficacy against certain nerve agents, e.g. soman and cyclosarin. In the past decades, numerous new oximes were synthesized and tested for their antidotal efficacy. The available data indicate that two Hagedorn oximes, HI 6 and HLo 7, are promising antidotes against various nerve agents. The efficacy of antidotes against nerve agent poisoning cannot be investigated in humans for ethical reasons. Therefore, it is necessary to use surrogate parameters for the evaluation of oxime efficacy. Reactivation of inhibited AChE is considered to be the main mechanism of action of oximes. Clinical data indicate that changes in erythrocyte AChE activity correlate to neuromuscular function indicating that interactions between AChE, inhibitor and oximes can be investigated in vitro with human erythrocyte AChE. Different theoretical models were used for the evaluation of reactivating efficacy of oximes with nerve agent-inhibited human AChE and for estimating effective oxime concentrations. The calculations demonstrate the marked differences between oximes in dependence of the inhibitor and provide a basis for the estimation of the required oxime dose as well as of dosing intervals. PMID- 16266696 TI - Solubilization and stabilization of beta-carotene in niosomes: delivery to cultured cells. AB - Carotenoids exhibit preventive effects against major diseases, including cancer and atherosclerosis. However, experimental studies on carotenoid functions in cultured cells are limited by the absence of an adequate method of solubilizing carotenoids, since they are unstable when exposed to light or oxygen and highly hydrophobic. In this study, we developed a niosomal formulation, consisting of non-ionic surfactants and cholesterol, which both solubilized and stabilized beta carotene and that allowed to deliver it to cultured cells at concentrations spanning the range of physiological levels. beta-Carotene contained in niosomes was highly resistant to sunlight, high temperatures and oxidative stress induced by different sources of free radicals. The carotenoid was extremely stable in culture medium up to 96 h. Moreover, it was easily taken up by both immortalized and transformed cells at carotenoid concentrations which ranged from 0.1 to 2 microM. Therefore, niosomes provide a convenient, nontoxic and inexpensive vehicle for beta-carotene in cell culture. PMID- 16266697 TI - Catecholamine analysis with microcolumn LC-peroxyoxalate chemiluminescence reaction detection. AB - BACKGROUND: Plasma catecholamines (CAs) are widely used as an index of sympathetic nervous system activity. In addition, CAs are known to be metabolized by catechol-O-methyltransferase (COMT) to produce their 3-O-methyl metabolites. We previously established a sensitive determination method of CAs and their 3-O methyl metabolites using HPLC-peroxyoxalate chemiluminescence (POCL) reaction detection system. In this study, a microcolumn (100 x 1.0 mm I.D.) was used for separation to obtain higher sensitivity and shorter analysis time. METHODS: The system included automated precolumn ion-exchange extraction of amines, followed by separation on an ODS column, coulometric oxidation, fluorescence derivatization with ethylenediamine, and finally POCL reaction detection. RESULTS: The detection limits for CAs and their 3-O-methyl metabolites were 0.3 2.0 fmol. The analysis time was about 35 min, about half that of previously reported results. The method developed was used in monitoring changes in CAs and 3-O-methyl metabolite concentrations in human plasma during exercise. CONCLUSION: The simultaneous determination method for concentrations of CAs and their 3-O methyl metabolites in human plasma was developed using micro LC-peroxyoxalate chemiluminescence detection. We were successful in quantitating the changes in plasma CAs and their 3-O-methyl metabolites during exercise. PMID- 16266698 TI - Oxidative status in patients submitted to conization and radiation treatments for uterine cervix neoplasia. AB - BACKGROUND: Cervical cancer is a major cause of morbidity among women. We investigated the treatment effect on oxidative status from patients submitted to radiotherapy or conization surgery to high-grade SIL (squamous intraepithelial lesion) treatment, and oxidative profile from patients newly diagnosed for uterine cervix cancer, without treatment. METHODS: We determined the catalase activity in blood, reduced glutathione (GSH) in plasma, TBARS and protein carbonyl content from serum samples of the patients. RESULTS: The catalase activity, GSH levels, TBARS and protein carbonyl content had no statistical differences related to the controls, neither when the 2 treatments were compared, possibly because the antioxidant defense may be acting in the first period of the neoplasic transformation, and maybe indicating a possible arrest of the tumor cells caused by the efficiency of the treatments. In the non-treated patients, TBARS and protein carbonyl contents, GSH levels and catalase activity were shown to be increased comparing with the treated patients and compared with the controls indicating an tumor effect on oxidative profile, and the antioxidant activity been increased in the beginning of the tumor development. CONCLUSIONS: We suggest that the treatments were efficient in arrest of the tumor. PMID- 16266699 TI - Attack behaviors in mice: from factorial structure to quantitative trait loci mapping. AB - The emergence or non-emergence of attack behavior results from interaction between the genotype and the conditions under which the mice are tested. Inbred mice of the same strain reared or housed under conditions do not react the same way; reactions also vary according to the place selected for testing and the different opponents. A factor analysis showed that the attack behavior in non isolated males, tested in neutral area covaried with high testosterone and steroid sulfatase and low brain 5-hydroxytriptamine (5-HT), beta-endorphin and Adrenocorticotropic Hormone (ACTH) concentration, whereas, for isolated males tested in their own housing cage, it covaried with high testosterone activity and low brain 5-HT concentration. A wide genome scan was performed with two independent populations derived from C57BL/6J and NZB/BlNJ, each being reared, housed and tested under highly contrasting conditions, as described above, and confronted with A/J standard males. Common Quantitative Trait Loci emerged for two rearing/testing conditions. For rattling latency we detected Quantitative Trait Loci on Mus musculus chromosome 8 (MMU8) (at 44, LOD score=3.51 and 47 cM, LOD score=6.22, for the first and the second conditions) and on MMU12 (at 39 cM, LOD score=3.69 and at 41 cM, LOD score=2.99, respectively). For the number of attacks, Quantitative Trait Loci were common: on MMU11 at 39 cM LOD score=4.51 and 45 cM, LOD score=3.05, respectively, and on MMU12 (17 cM, LOD score=2.71 and 24 cM, LOD score=3.10). The steroid sulfatase gene (Sts), located on the X-Y pairing region, was linked, but only in non-isolated males, tested in neutral area for rattling latency, first attack latency, and number of attacks (LOD scores=4.9, 4.79 and 3.57, respectively). We found also that the Quantitative Trait Locus encompassing Sts region interacted with other Quantitative Trait Loci. These results indicate that attack behavior measured in different rearing and testing conditions have different biological and genetic correlates. This suggests that further explorations should be done with standardized tests and, in addition, with a wide range of tests, so as to gain an understanding of the true impact of genes or pharmacological treatments on specific categories of aggressive behavior. PMID- 16266700 TI - New achievements and pharmacotherapeutic approaches in the treatment of alcohol dependence. AB - The treatment of alcohol dependence mainly consists of psychological, social, and pharmacotherapeutic interventions aiming to reduce physical withdrawal, craving, and alcohol relapse. During the last years, it has become increasingly clear that adjuvant pharmacotherapy is efficacious especially in rehabilitation programs for alcohol dependent patients. The development of alcohol dependence seems to involve adaptive changes in amino acid neurotransmitter systems, stimulation of dopamine and opioid peptide systems, and changes in serotonergic activity. Disulfiram, naltrexone and acamprosate are approved treatments for the management of abstinence maintenance treatment. New compounds are under investigation. This review discusses the neurobiological basis of alcohol addiction, pharmacological targets for relapse prevention treatment and pre-clinical and clinical results with the most promising drugs. PMID- 16266701 TI - Effect of mast cells depletion on the failure of neutrophil migration during sepsis. AB - The possible role of mast cell in neutrophil migration failure during sepsis was examined in a polymicrobial sepsis model in mice. Mast cells were depleted by compound 48/80 or lysed by distilled water, both preventing the neutrophil migration failure. This phenomenon was accompanied by reduction of bacteria in the peritoneal cavity and blood, serum tumor necrosis factor alpha (TNF-alpha), interleukin 1beta (IL-1beta) and nitrate (NO3) and by an increase in mice survival rate. Neither neutrophil migration failure nor significant mortality was observed when lethal inoculum was injected into the air-pouch model, a cavity poorly populated by mast cells. Confirming that neutrophil migration failure is a phenomenon induced by systemic circulating mediators, it was observed that i.p. administration of lethal inoculum induced a neutrophil migration failure to the air pouch inoculated with non-lethal bacterial challenge. These results suggest that mast cells have a key role in the genesis of neutrophil migration failure, and, consequently, contribute to the systemic inflammatory response and mortality in severe sepsis. PMID- 16266702 TI - Reduced capacitative calcium entry in the mesenteric vascular bed of bile duct ligated rats. AB - In this work, we analyzed the interaction of nitric oxide (NO) with some of the mechanisms known to regulate intracellular calcium levels in order to gain insight into the mechanisms responsible for the reduced vascular pressor response to vasoconstrictors observed in an experimental model of liver cirrhosis. Specifically, we hypothesized that the entry of calcium through capacitative channels is defective in this model. The experiments were performed with isolated, Krebs-perfused and de-endothelialized mesenteric arterial bed of rats with bile duct ligation (4 weeks) and their controls. Pretreatment with thapsigargin to inhibit calcium uptake into sarcoplasmic reticulum potentiated the pressor responses to methoxamine, but the response of the cirrhotic vessels was significantly lower than that of the controls. Under the same conditions, perfusion of the mesenteries with zero calcium-Krebs resulted in lower pressor responses to methoxamine, especially in the mesenteries of the bile duct-ligated rats. To specifically analyze the entry of calcium through store-operated calcium channels, the pressor response to the addition of calcium was studied in mesenteries perfused with zero calcium-Krebs and in the presence of thapsigargin. Again, the response of the cirrhotic mesenteric beds was significantly lower than that of the control vessels. Under all these experimental conditions, the differences between control and cirrhotic responses were abolished by pretreatment with the NO synthesis inhibitor N(w)-nitro-L-arginine (NNA). These results indicate that, in the mesenteric bed of bile duct-ligated rats, an excess of nitric oxide interferes with the release of calcium from thapsigargin sensitive internal stores and also reduces the capacitative entry of calcium into vascular muscular cells induced by the depletion of calcium from internal stores. This mechanism may have an important role in the reduced pressor response observed in the mesenteric vascular bed in cirrhosis. PMID- 16266703 TI - Rhesus monkey alpha7 nicotinic acetylcholine receptors: comparisons to human alpha7 receptors expressed in Xenopus oocytes. AB - An alpha7 nicotinic acetylcholine receptor sequence was cloned from Rhesus monkey (Macaca mulatta). This clone differs from the mature human alpha7 nicotinic acetylcholine receptor in only four amino acids, two of which are in the extracellular domain. The monkey alpha7 nicotinic receptor was characterized in regard to its functional responses to acetylcholine, choline, cytisine, and the experimental alpha7-selective agonists 4OH-GTS-21, TC-1698, and AR-R17779. For all of these agonists, the EC(50) for activation of monkey receptors was uniformly higher than for human receptors. In contrast, the potencies of mecamylamine and MLA for inhibiting monkey and human alpha7 were comparable. Acetylcholine and 4OH-GTS-21 were used to probe the significance of the single point differences in the extracellular domain. Mutants with the two different amino acids in the extracellular domain of the monkey receptor changed to the corresponding sequence of the human receptor had responses to these agonists that were not significantly different in EC(50) from wild-type human alpha7 nicotinic receptors. Monkey alpha7 nicotinic receptors have a serine at residue 171, while the human receptors have an asparagine at this site. Monkey S171N mutants were more like human alpha7 nicotinic receptors, while mutations at the other site (K186R) had relatively little effect. These experiments point toward the basic utility of the monkey receptor as a model for the human alpha7 nicotinic receptor, albeit with the caveat that these receptors will vary in their agonist concentration dependency. They also point to the potential importance of a newly identified sequence element for modeling the specific amino acids involved with receptor activation. PMID- 16266704 TI - Chronic spinal cord injury induced changes in the responses of thalamic neurons. AB - Sensory disturbances following spinal cord injury (SCI) include chronic pain, which is often localized at spinal levels just rostral to the lesion (referred to as at-level neuropathic pain) and not effectively relieved by traditional treatments. In the present study, a clinically relevant spinal contusion injury was made at the spinal T8 level in 11 deeply anesthetized male rats. Behavioral testing just prior to terminal electrophysiological experiments (done at 30 days post-injury) demonstrated at-level sensitivity to touching the trunk (i.e., allodynia) in 64% of the animals. Electrophysiological data (urethane anesthesia) were obtained for 218 single somatovisceral convergent neurons that were located throughout 12 subregions of the thalamus. In total, 90% (197 of 218) responded to noxious at-level pinch, compared to 52% for pinching the dorsal trunk at the same level in uninjured controls (our previously published data--recorded from 133 total neurons). In addition, 33% of the total neurons tested also responded to gentle touch (dorsal trunk) versus 9% in controls. A comparison of electrophysiological and behavioral data for each individual animal reveals novel tactile neuronal responses within ventral and posterior thalamic subnuclei for those rats showing signs of at-level allodynia. These data suggest that neurons in specific regions of thalamus undergo significant changes in responsiveness following severe chronic SCI. The observed plasticity and ensuing hypersensitivity are likely part of the central reorganization producing the multitude of sensory disturbances that surface following SCI. PMID- 16266705 TI - Molecular and developmental analyses of thyroid hormone receptor function in Xenopus laevis, the African clawed frog. AB - The current review focuses on the molecular mechanisms and developmental roles of thyroid hormone receptors (TRs) in gene regulation and metamorphosis in Xenopus laevis and discusses implications for TR function in vertebrate development and diversity. Questions addressed are: (1) what are the molecular mechanisms of gene regulation by TR, (2) what are the developmental roles of TR in mediating the thyroid hormone (TH) signal, (3) what are the roles of the different TR isoforms, and (4) how do changes in these molecular and developmental mechanisms affect evolution? Even though detailed knowledge of molecular mechanisms of TR-mediated gene regulation is available from in vitro studies, relatively little is known about how TR functions in development in vivo. Studies on TR function during frog metamorphosis are leading the way toward bridging the gap between in vitro and in vivo studies. In particular, a dual function model for the role of TR in metamorphosis has been proposed and investigated. In this model, TRs repress genes allowing tadpole growth in the absence of TH during premetamorphosis and activate genes important for metamorphosis when TH is present. Despite the lack of metamorphosis in most other vertebrates, TR has important functions in development across vertebrates. The underlying molecular mechanisms of TR in gene regulation are conserved through evolution, so other mechanisms involving TH target genes and TH tissue-sensitivity and dependence underlie differences in role of TR across vertebrates. Continued analysis of molecular and developmental roles of TR in X. laevis will provide the basis for understanding how TR functions in gene regulation in vivo across vertebrates and how TR is involved in the generation of evolutionary diversity. PMID- 16266706 TI - The attitudes of two groups of South African women towards mode of delivery. AB - OBJECTIVE: To determine whether 2 specific groups of women prefer vaginal or abdominal delivery. METHODS: Forty-six black and 54 colored (of mixed race, the predominant population group) primigravidas and 59 black and 62 colored multigravidas were interviewed between April 2003 and February 2004 at Tygerberg Hospital, Tygerberg, South Africa. RESULTS: Among primigravidas, 89% of black women and 83% of colored women preferred vaginal delivery. Among the multigravidas, 86% of black women and 79% of colored women preferred vaginal delivery. Most women believed that they would recover more quickly (65%) and with less pain (32%) after vaginal delivery. Forty-one percent of all women viewed episiotomy as the greatest disadvantage of vaginal delivery, while 43% were unaware of any advantage to cesarean delivery. CONCLUSION: Most colored and black women preferred vaginal over cesarean delivery. A challenge to healthcare personnel is to educate women about both routes. PMID- 16266707 TI - Thermodynamic analysis of the heterogenous binding sites of molecularly imprinted polymers. AB - The thermodynamic interactions of two polymers, one Fmoc-L-Trp-imprinted (MIP), the other one an unimprinted reference (NIP), with the two Fmoc-tryptophan enantiomers were studied by frontal analysis, which allows accurate measurements of the adsorption isotherms. These isotherms were acquired at temperatures of 40, 50, 60, and 70 degrees C, for sample concentrations ranging between 0.005 and 40 mM. The mobile phase used was acetonitrile with one percent acetic acid as an organic modifier. Within the measured concentration ranges, the tri-Langmuir isotherm model accounts best for the isotherm data of both enantiomers on the MIP, the bi-Langmuir model for the isotherm data of Fmoc-L-Trp on the NIP. These isotherm models were selected using three independent processes: statistical tests on the results from regression of the isotherm data to different isotherm models; calculation of the affinity energy distribution from the raw isotherm data; comparison of the experimental and the calculated band profiles. The isotherm parameters obtained from these best selected isotherm models showed that the enantiomeric selectivity does not change significantly with temperature, while the affinity of the substrates for both the MIP and the NIP decrease considerably with increasing temperatures. These temperature effects on the binding performance of the MIP were clarified by considering the thermodynamic functions (i.e., the standard molar Gibbs free energy, the standard molar entropy of adsorption, and the standard molar enthalpy of adsorption) for each identified type of adsorption sites, derived from the Van't Hoff equation. This showed that the entropy of transfer of Fmoc-L-Trp from the mobile to the MIP stationary phase is the dominant driving force for the selective adsorption of Fmoc-L-Trp onto the enantioselective binding sites. This entropy does not change significantly with increasing temperatures from 40 to 70 degrees C. PMID- 16266708 TI - Determination of trace concentrations of bromophenols in water using purge-and trap after in situ acetylation. AB - An analytical method that enables detection and quantification of bromophenols (BPs) at taste threshold concentrations (2,6-DBP: 0.5 ng/L) was developed. This method involves conversion of the BPs to their acetates, followed by isolation of the acetates by a modified purge-and-trap procedure, and analysis by gas chromatography-mass spectrometry (GC-MS) in selected ion monitoring (SIM) mode. Bromophenyl acetates were synthesized so that each of the two steps in the method could be developed and optimised in isolation to the other. Deuterated BPs (phenol-d5, 2-BP-d4, 4-BP-d2, 2,6-DBP-d3, 2,4-DBP-d3 and 2,4,6-TBP-d2) were synthesized to enable quantification of analytes using the deuterated analogues of analytes as internal standards. This method allowed quantification of BPs at concentrations ranging from the detection limits (3 ng/L for phenol and 0.1-0.5 ng/L for each of the BPs) to 1000 ng/L for each analyte, with repeatabilities of < or =14% (RSD) for concentrations of 1 ng/L and < or =9% (RSD) for concentrations of 10-1000 ng/L, with recoveries ranging from 91 to 97%. PMID- 16266709 TI - Studies of the resolution of racemic 1,1'-bi-2-naphthol with a dipeptide chiral selector identified from a small library. AB - Several new stationary phases were prepared to study the structure-activity relationship of the chiral resolution of racemic 1,1'-bi-2-naphthol with a modified dipeptide Asn-Asn selector. The number of amino acid, the side chain protecting groups of the amino acid, and the Fmoc end-capping group all proved important for enantioselectivity. The linker also influenced enantioselectivity. Influence of the length of the linker appears to be related to the accessibility of chiral selectors. The bond through which the selector is attached to the linker proved important. Based on these results, it is postulated that hydrogen bonding interactions between one side chain amide group of one Asn and the oxygen on the backbone of another Asn with the two hydroxyl groups of the analyte play an important role in the resolution of racemic 1,1'-bi-2-naphthol with the modified dipeptide Asn-Asn selector. PMID- 16266710 TI - High-throughput screening of protein binding by equilibrium dialysis combined with liquid chromatography and mass spectrometry. AB - A new approach for screening plasma protein binding is presented. The method is based on equilibrium dialysis combined with rapid generic LC-MS bioanalysis by using a sample pooling approach enabling high-throughput screening of protein binding in the drug discovery phase. The method is evaluated by a comparison of measured unbound free fractions f(u) (%) between single and pooled compounds for a test set of structurally diverse compounds with a wide range of unbound fractions. Test compounds include 1 acidic and 10 basic drug standards along with 36 new chemical entities. A good correlation (R2>0.95) of f(u) (%) between the single and pooled compounds is found, suggesting that at least 10 compounds can be simultaneously measured with acceptable accuracy. A simplified drug-protein binding model is applied to calculate the f(u) (%) of drugs at various drug and protein concentrations and this is applied to elucidate the applicability of the sample pooling approach from a theoretical standpoint. Moreover, pH shifts in the plasma were observed after dialysis when using different types of buffers and the impact of that on the f(u) is illustrated in association with their physicochemical properties, in particular the ionization state of compounds by the profile of effective mobility as a function of pH. A new buffer is proposed being able to minimize the pH shift of plasma during the dialysis. In addition, the application of the proposed buffer does not necessarily require adjusting plasma pH before the dialysis and utilizing a CO2 incubator during the dialysis. The effect of the ionic strengths of different buffers on MS signals is investigated with regard to ion suppression. The sample pooling method not only significantly reduces the plasma volume required but also the number of bioanalysis samples as compared to the single compound measurements by a conventional approach. The new proposed approach is especially beneficial for measuring in vitro protein binding in matrices such as mouse plasma where plasma is available only in limited amounts. The current new development will facilitate the drug discovery process by more rapidly assessing the protein binding potential of drug candidates. PMID- 16266711 TI - Improved solvent-free microwave extraction of essential oil from dried Cuminum cyminum L. and Zanthoxylum bungeanum Maxim. AB - Solvent-free microwave extraction (SFME) is a recently developed green technique which is performed in atmospheric conditions without adding any solvent or water. SFME has already been applied to extraction of essential oil from fresh plant materials or dried materials prior moistened. The essential oil is evaporated by the in situ water in the plant materials. In this paper, it was observed that an improved SFME, in which a kind of microwave absorption solid medium, such as carbonyl iron powders (CIP), was added and mixed with the sample, can be applied to extraction of essential oil from the dried plant materials without any pretreatment. Because the microwave absorption capacity of CIP is much better than that of water, the extraction time while using the improved SFME is no more than 30 min using a microwave power of 85 W. Compared to the conventional SFME, the advantages of improved SFME were to speed up the extraction rate and need no pretreatment. Improved SFME has been compared with conventional SFME, microwave assisted hydrodistillation (MAHD) and conventional hydrodistillation (HD) for the extraction of essential oil from dried Cuminum cyminum L. and Zanthoxylum bungeanum Maxim. By using GC-MS system the compositions of essential oil extracted by applying four kinds of extraction methods were identified. There was no obvious difference in the quality of essential oils obtained by the four kinds of extraction methods. PMID- 16266712 TI - Analysis of phenols in water by high-performance liquid chromatography using coumarin-6-sulfonyl chloride as a fluorogenic precolumn label. AB - A simple, sensitive and rapid reversed-phase high-performance liquid chromatography (RP-HPLC) method is proposed for the analysis of some environmentally important phenols in water. The use of coumarin-6-sulphonyl chloride (C6SCl) as a fluorescence-labeling reagent has been investigated. The compound reacts with phenols within 20 min under mild conditions (ambient temperature, pH 9.0) to give sulphonates that can be separated by RP-HPLC employing fluorescence detection at lambda(ex) = 360 and lambda(em) = 460 nm. The optimum conditions for fluorescence, derivatization and chromatographic separation have been established and detection limits in the range 0.1-0.9 microg l(-1) were obtained for the studied compounds. The calibration curves were linear for the range 6-200 microg l(-1) for phenol, 3-200 microg l(-1) for 2 chlorophenol, 4-chlorophenol and 2,3,5-trichlorophenol and for the range of 3-100 microg l(-1) for 2,3-dichlorophenol and 3,5-dichlorophenol. The practical applicability of the method to environmental samples was demonstrated by analyzing drinking and industrial water samples spiked with the phenolic compounds. PMID- 16266713 TI - Unusual resolution of N-(3,5-dinitrobenzoyl)-alpha-amino acids on a chiral stationary phase based on (+)-(18-crown-6)-2,3,11,12-tetracarboxylic acid. AB - While HPLC chiral stationary phases (CSPs) based on chiral crown ethers have been known useful for the resolution of only racemic primary amino compounds or some secondary amino compounds, in this study, we first demonstrated that the CSP based on (+)-(18-crown-6)-2,3,11,12-tetracarboxylic acid is also useful for the resolution of N-benzoyl-alpha-amino acids, which do not contain a primary or secondary amino group. Especially, N-(3,5-dinitrobenzoyl)-alpha-amino acids were resolved better than corresponding N-(3-nitrobenzoyl)- or N-benzoyl-alpha-amino acids, the separation (alpha) and the resolution factors (R(S)) for the resolution of eight N-(3,5-dinitrobenzoyl)-alpha-amino acids being in the range of 1.06-1.81 and 0.54-2.81, respectively. The optimum mobile phase condition was the mixture of acetic acid-triethylamine-acetonitrile with the ratio of 0.05/0.25/100 (v/v/v). PMID- 16266714 TI - Stir bar sorptive extraction for the determination of volatile compounds in oak aged wines. AB - Stir bar sorptive extraction (SBSE) followed by a thermal desorption-gas chromatography-mass spectrometry analysis has been optimized for the determination of 13 of the most important oak volatiles in wine samples, all in a single run. The stir bar sorptive extraction method was optimized in terms of temperature, time, pH and NaCl addition, and the best results were obtained after stirring the wine sample with the polydimethylsiloxane stir bar during 90min at room temperature. The addition of sodium chloride did not enhance the volatile extraction. The method proposed showed good linearity over the concentration range tested, with correlation coefficients ranging from 0.92 to 0.99 for all the analytes. The reproducibility and repeatability of the method was estimated between 0.11 and 5.45%. The detection and quantification limits of all analytes were lower than their respective olfactory threshold values, and, most importantly, no artifacts have been observed during the analysis as described in most extractions using other current methodologies. PMID- 16266716 TI - Structural evolution of self-expanding arrays of charged particulates. AB - Self-expansion patterns of unconstrained assemblies of charged particulates are simulated by solution of their individual trajectories. The general behaviour of these systems is considered regarding their expansion shape and structure. As the particulates cannot be described, in general, in terms of massless charged entities, the complete equation of motion, inclusive of the inertial and other size effects, must be applied to each and every member of the assembly. It is shown that irrespective of the initial position of the particulates and the time dependent shape of the assembly, when expanding in free space or else the particulates are identical in size, shape and mass, they self-expand asymptotically into a circular or spherical shape with an inner structure that tends to uniformity. This behaviour persists irrespective of the size and charge level of the particulates, or whether they form a single or multiple separate groups in one, two and three dimensions. In this context, ionic gaseous assemblies that fit into the realm of continua, are included. Two- and three dimensional examples of simulation outputs for different particulate assemblies, illustrate typical self-expansion patterns. Internal structures that evolve in two-dimensional self-expanding arrays are shown to be different compared to those obtained in three dimensions. These simulations show that models of particle capture by random self-expanding arrays of charged particulates, may lack physical grounds, as they contradict the asymptotic mode of uniform and ordered self-expansion that is expected from the array. PMID- 16266715 TI - Sample preparation and gas chromatography of primary fatty acid amides. AB - A method for the isolation of bio-active primary fatty acid amides (PFAM's) from total lipid extract by solid-phase extraction (SPE) was developed and validated. The lowest mass of amide to be loaded and recovered by this method was detected as 0.5 microg using 500 mg of normal phase adsorbent. The isolated PFAM's were separated and quantified by GC/MS and percent recoveries were calculated. An HP 5MS column was able to provide base line separation between the saturated and unsaturated PFAM's whereas clear resolution between geometric and positional isomers having the same number of carbons was obtained using a BPX70 column. The separated amides were all 18 carbon analogs of cis-9-octadecenoamide (oleamide). Detection limits in the single ion monitoring mode were found to be on the order of 10 pg in a 1 microl injection. Solid phase extraction of amides from total lipid extract before GC/MS analysis provides clean detection and interference free analysis. PMID- 16266717 TI - Compressibility, the measurement of surface tension, and particle size in molecular or nuclear matter. AB - It is shown that the interface shrinkage resulting from the capillary pressure difference between both sides of a curved interface is the product of a "standard shrinkage"kappagamma (kappa is the isothermal compressibility, gamma the interfacial tension) by a dimensionless factor that depends only on the shape of the sample of matter under study. The behaviour of the standard shrinkage in the critical domain shows that it cannot be a measure of the thickness of the liquid vapour interface in that domain. The standard shrinkage of classical liquids somewhat above triple point is usually near to 0.048 v(c)(1/3) (v(c) is the critical molecular volume); exceptions to this rule are discussed. The variation of the standard shrinkage along the liquid-vapour coexistence curves of water and argon is presented; the effect of the interface shrinkage on the measured surface tension of liquids can become important within about 15% of the critical temperature. The standard shrinkage of solids is less than that of the corresponding liquids, and is of no consequence when measuring the surface tension of solids. The standard shrinkage of the nuclear fluid is 0.23 fm=0.09 v(c)(1/3). The saturation density of infinite nuclear matter is about 9% less than its value in atomic nuclei, and a term proportional to A(1/3) (A is the mass number) must be added to the nuclear binding energy formula. PMID- 16266718 TI - Autocrine activation of DNA synthesis in prothoracic gland cells of the silkworm, Bombyx mori. AB - Autocrine activation of DNA synthesis in prothoracic gland cells in last instar larvae of the silkworm, Bombyx mori, was studied using both a long-term in vitro organ culture system and immunocytochemical labeling with 5-bromo-2'-deoxyuridine (BrdU). When prothoracic glands were incubated in a small volume of culture medium (10 microl/gland), the numbers of DNA-synthesizing cells per gland increased significantly, and DNA synthesis was stimulated less by hemolymph, as compared with glands incubated in a large volume (50 microl/gland). Moreover, glands cultured in groups (6 glands per group in a 50-microl drop) also resulted in much higher levels of DNA synthesis than those cultured individually in a 50 microl drop. The mechanism by which alternation of the volume of the incubation medium results in changes in the levels of DNA synthesis was further examined. When prothoracic glands were incubated in medium (50-microl drop per gland) that was preconditioned with glands (in a 10-microl drop individually), a dramatic increase in DNA synthesis activity was also observed, indicating that prothoracic glands may release a factor that stimulates their own DNA synthesis. The growth promoting factor was further characterized and it was found that the factor is heat stable, and its molecular weight was estimated to be between 1,000 and 3,000 Da. Moreover, the factor also stimulated corpus allatum cell DNA synthesis in vitro. Injection of concentrated putative growth-promoting factor into day 4 last instar-ligated larvae greatly increased cell DNA synthesis of the prothoracic glands, indicating the in vivo function of the present autocrine factor. PMID- 16266720 TI - Increased serum-GFAP in patients with severe traumatic brain injury is related to outcome. AB - OBJECTIVES: Several studies have established the relevance of S-100 in blood as a marker of brain damage after traumatic brain injury. However, a more specific marker is required and glial fibrillary acidic protein (GFAP) is considered to be a good candidate. METHODS: In order to assess the increase of GFAP in serum (s GFAP) after a severe traumatic brain injury (TBI) we collected daily serum samples from 59 patients with severe TBI starting on the day of the trauma. S GFAP was measured using a sandwich ELISA. The Glasgow outcome scale (GOS) assessed outcome after 1 year. RESULTS: All but one patient had maximal s-GFAP values above the laboratory reference value (median increased 10-fold). The highest detected levels were seen during the first days after TBI and then decreased gradually. Patients with unfavourable outcome had significantly (p<0.001) higher maximal s-GFAP values in the acute phase compared with patients with favourable outcome. All patients (n=5) with s-GFAP>15.04 microg /L died (reference level<0.15 microg/L). We found no significant difference in the maximal s-GFAP levels of patients with isolated brain injury in comparison with patients with multiple traumas. CONCLUSION: Serum-GFAP is increased during the first days after a severe traumatic brain injury and related to clinical outcome. PMID- 16266721 TI - Action of imipramine on activated ATP-sensitive K(+) channels in interstitial cells of Cajal from murine small intestine. AB - Tricyclic antidepressants have been widely used for the treatment of depression and as a therapeutic agent for the altered gastrointestinal (GI) motility of irritable bowel syndrome (IBS). The aim of this study was to clarify whether antidepressants directly modulate pacemaker currents in cultured interstitial cells of Cajal (ICC). We used the whole-cell patch-clamp techniques at 30 degrees C in cultured ICC from the mouse small intestine. Treatment of pinacidil, an ATP sensitive K(+) channel opener, in the ICC using the current clamping mode, produced hyperpolarization of the membrane potential and decreased the amplitude of the pacemaker potentials. With the voltage clamp mode, we observed a decrease in the frequency and amplitude of pacemaker currents and increases in the resting outward currents. These effects of pinacidil on pacemaker potentials and currents were completely suppressed by glibenclamide, an ATP-sensitive K(+) channel blocker. Also, with the current clamp mode, imipramine blocked the affect of pinacidil on the pacemaker potentials. Observations of the voltage clamp mode with imipramine, desipramine and amitryptyline suppressed the action of pinacidil in the ICC. Next, we examined whether protein kinase C (PKC) and the G protein are involved in the action of imipramine on pinacidil induced pacemaker current inhibition. We used chelerythrine, a potent PKC inhibitor and GDPbetaS, a nonhydrolyzable guanosine 5-diphosphate (GDP) analogue that permanently inactivates GTP-binding proteins. We found that pretreatment with chelerythrine and intracellular application of GDPbetaS had no influence on the blocking action of imipramine on inhibited pacemaker currents by pinacidil. We conclude that imipramine inhibited the activated ATP-sensitive K(+) channels in ICC. This action does not appear to be mediated through the G protein and protein kinase C. Furthermore, this study may suggest another possible mechanism for tricyclic antidepressants related modulation of GI motility. PMID- 16266719 TI - Divergent evolution within protein superfolds inferred from profile-based phylogenetics. AB - Many dissimilar protein sequences fold into similar structures. A central and persistent challenge facing protein structural analysis is the discrimination between homology and convergence for structurally similar domains that lack significant sequence similarity. Classic examples are the OB-fold and SH3 domains, both small, modular beta-barrel protein superfolds. The similarities among these domains have variously been attributed to common descent or to convergent evolution. Using a sequence profile-based phylogenetic technique, we analyzed all structurally characterized OB-fold, SH3, and PDZ domains with less than 40% mutual sequence identity. An all-against-all, profile-versus-profile analysis of these domains revealed many previously undetectable significant interrelationships. The matrices of scores were used to infer phylogenies based on our derivation of the relationships between sequence similarity E-values and evolutionary distances. The resulting clades of domains correlate remarkably well with biological function, as opposed to structural similarity, indicating that the functionally distinct sub-families within these superfolds are homologous. This method extends phylogenetics into the challenging "twilight zone" of sequence similarity, providing the first objective resolution of deep evolutionary relationships among distant protein families. PMID- 16266722 TI - Nicotine and lipopolysaccharide stimulate the formation of osteoclast-like cells by increasing macrophage colony-stimulating factor and prostaglandin E2 production by osteoblasts. AB - Several studies have indicated that one of the causes of alveolar bone destruction with periodontitis is lipopolysaccharide (LPS) from the cell wall of Gram-negative bacteria in plaque and that tobacco smoking may be an important risk factor for the development and severity of periodontitis. The present study was undertaken to determine the effect of nicotine and LPS on the expression of macrophage colony-stimulating factor (M-CSF), osteoprotegerin (OPG), and prostaglandin E2 (PGE2) in osteoblasts, and the indirect effect of nicotine and LPS on the formation of osteoclast-like cells. Saos-2 cells were cultured with 10(-3) M nicotine, or 1 or 10 microg/ml LPS and 10(-3) M nicotine, for up to 14 days. The gene and protein expression of M-CSF and OPG were determined using real time PCR and ELISA, respectively. PGE2 expression was determined using ELISA. The formation of osteoclast-like cells was estimated using tartrate-resistant acid phosphatase (TRAP) staining of osteoclast precursors in culture with conditioned medium from nicotine and LPS-treated Saos-2 cells and the soluble receptor activator of NF-kappaB ligand (RANKL). M-CSF and PGE2 expression increased markedly in cells cultured with nicotine and LPS compared with those cultured with nicotine alone. OPG expression increased in the initial stages of culture with nicotine and LPS but decreased in the later stages of culture. The conditioned medium containing M-CSF and PGE2 produced by nicotine and LPS-treated Saos-2 cells with soluble RANKL increased the TRAP staining of osteoclast precursors compared with that produced by nicotine treatment alone. These results suggest that nicotine and LPS stimulate the formation of osteoclast-like cells via an increase in M-CSF and PGE2 production and that the stimulation is greater than with nicotine treatment alone. PMID- 16266723 TI - Partial agonistic effect of 9-hydroxycorynantheidine on mu-opioid receptor in the guinea-pig ileum. AB - Mitragynine is an indole alkaloid isolated from the Thai medicinal plant Mitragyna speciosa that is reported to have opioid agonistic properties. The 9 demethyl analogue of mitragynine, 9-hydroxycorynantheidine, is synthesized from mitragynine. 9-Hydroxycorynantheidine inhibited electrically stimulated guinea pig ileum contraction, but its maximum inhibition was weaker than that of mitragynine and its effect was antagonized by naloxone, suggesting that 9 hydroxycorynantheidine possesses partial agonist properties on opioid receptors. Receptor binding assays revealed that 9-hydroxycorynantheidine has high affinity for mu-opioid receptors. In an assay of the guinea-pig ileum, naloxone shifted the concentration-response curves for [D-Ala(2), N-MePhe(4), Gly-ol(5)] enkephalin (DAMGO), (5alpha,7alpha,8beta)-(+)-N-Methyl-N-[7-(1-pyrrolidinyl)-1 oxaspiro[4.5]dec-8-yl]-benzeneacetamide (U69593) and 9-hydroxycorynantheidine to the right in a competitive manner. The pA(2) values of naloxone against 9 hydroxycorynantheidine and DAMGO were very similar, but not that against U69593. As indicated by the two assay systems, the opioid effect of 9 hydroxycorynantheidine is selective for the mu-opioid receptor. 9 Hydroxycorynantheidine shifted the concentration-response curve for DAMGO slightly to the right. Pretreatment with the mu-opioid selective and irreversible antagonist beta-funaltorexamine hydrochloride (beta-FNA) shifted the concentration-response curve for DAMGO to the right without affecting the maximum response. On the other hand, beta-FNA did not affect the curve for 9 hydroxycorynantheidine, but decreased the maximum response because of the lack of spare receptors. These studies suggest that 9-hydroxycorynantheidine has partial agonist properties on mu-opioid receptors in the guinea-pig ileum. PMID- 16266724 TI - Common carotid artery stiffness, cardiovascular function and lipid metabolism after menopause. AB - While cardiovascular disease is a major cause of death in elderly women, relatively little is known regarding the influence of menopause on atherogenesis. We tried to characterize postmenopausal changes in the arterial properties. A group of 72 postmenopausal women were classified into subgroups based on duration of the postmenopausal period (PMP): Group PM1 (1-2 years; n = 16), PM4 (2-6 years; n = 16), PM8 (6-10 years; n = 25), and PM12 (10-15 years; n = 15). The control group consisted of 24 volunteers with regular menstruation (PM0). The diameter pulse waveform and intima-media thickness (IMT) of the common carotid artery (CCA) was measured using a phase-locked echo tracking system coupled with B-mode ultrasonography. The stiffness index was calculated from the waveform and the systemic blood pressure. The cardiac contractile force and the cerebral perfusion were also estimated using the maximum incremental velocity (MIV) and the calculated blood flow, as well as the fasting lipid profile. When compared to control, significant and progressive increases were noted in total cholesterol and low density lipoprotein (PM1, PM4, PM8, PM12), IMT (PM8, PM12), and SI (PM1, PM4, PM8, PM12). Further significant and progressive reductions were noted in pulse amplitude of CCA diameter (PM1, PM4, PM8, PM12) and MIV and cerebral perfusion (PM8, PM12). The postmenopausal increase in CCA stiffness as well as lipid profile occurs earlier than the increase in IMT and may be a more sensitive predictor of disorder on arterial property. PMID- 16266725 TI - Prevention of kainic acid-induced changes in nitric oxide level and neuronal cell damage in the rat hippocampus by manganese complexes of curcumin and diacetylcurcumin. AB - Curcumin is a natural antioxidant isolated from the medicinal plant Curcuma longa Linn. We previously reported that manganese complexes of curcumin (Cp-Mn) and diacetylcurcumin (DiAc-Cp-Mn) exhibited potent superoxide dismutase (SOD)-like activity in an in vitro assay. Nitric oxide (NO) is a free radial playing a multifaceted role in the brain and its excessive production is known to induce neurotoxicity. Here, we examined the in vivo effect of Cp-Mn and DiAc-Cp-Mn on NO levels enhanced by kainic acid (KA) and L-arginine (L-Arg) in the hippocampi of awake rats using a microdialysis technique. Injection of KA (10 mg/kg, i.p.) and L-Arg (1000 mg/kg, i.p.) significantly increased the concentration of NO and Cp Mn and DiAc-Cp-Mn (50 mg/kg, i.p.) significantly reversed the effects of KA and L Arg without affecting the basal NO concentration. Following KA-induced seizures, severe neuronal cell damage was observed in the CA1 and CA3 subfields of hippocampal 3 days after KA administration. Pretreatment with Cp-Mn and DiAc-Cp Mn (50 mg/kg, i.p.) significantly attenuated KA-induced neuronal cell death in both CA1 and CA3 regions of rat hippocampus compared with vehicle control, and Cp Mn and DiAc-Cp-Mn showed more potent neuroprotective effect than their parent compounds, curcumin and diacetylcurcumin. These results suggest that Cp-Mn and DiAc-Cp-Mn protect against KA-induced neuronal cell death by suppression of KA induced increase in NO levels probably by their NO scavenging activity and antioxidative activity. Cp-Mn and DiAc-Cp-Mn have an advantage to be neuroprotective agents in the treatment of acute brain pathologies associated with NO-induced neurotoxicity and oxidative stress-induced neuronal damage such as epilepsy, stroke and traumatic brain injury. PMID- 16266726 TI - Effect of selenium-enriched malt on hepatocarcinogenesis, paraneoplastic syndrome and the hormones regulating blood glucose in rats treated by diethylnitrosamine. AB - 233 SD rats weighing 100 approximately 120 g were divided randomly into 6 groups. The animals in group I and group II received 0.1 mg/kg selenium in the form of sodium selenite only and served as the negative control and positive control, respectively. Animals in groups III, IV and V were fed with selenium as Se enriched malt supplemented diets (0.3, 1 and 3 mg/kg), and group VI with selenium by using sodium selenite supplemented diets (3 mg/kg). Animals of groups II approximately VI were induced hepatoma by diethylnitrosamine (100 mg/l) for 16 weeks, then drunk with sterilized water for 2 more weeks. Subsequently, the effects of Se-enriched malt and sodium selenite on hepatoma nodules, relative liver weight, the liver function indices including alanine aminotransferase (ALT), alkaline phosphatase (ALP), albumin (ALB), total bilirubin (TBIL), and the tumor markers, named as gamma-glutamyltranspeptidase (GGT), alpha-fetoprotein (AFP), insulin-like growth factor-II (IGF-II) were recorded. The calcium concentration, glucose content in plasma and values of the hormones regulating blood glucose, such as insulin, glucagons and thyroid hormones (3,5,3' tetraiodothyronine, T(3); 3,5,3'5'-tetraiodothyronine, T(4)) were observed as well. At the same time, the correlations between the concentration of plasma glucose and related hormones were also analyzed. The results indicated that Se enriched malt showed a better chemopreventive efficiency in decreasing the number of hepatoma nodules, relative liver weight and the contents of AFP, GGT, IGF-II, ALT, ALP and TBIL in the plasma, and delaying the descent of hormones in the serum, names as insulin, glucagons, T(3) and T(4) than those feeding with sodium selenite. Effect of Se-enriched malt excelled sodium selenite in the aspects of deadening the descent of glucose concentration in the plasma and the rise of calcium concentration in the serum of the rats with hepatoma induced by diethylnitrosamine. The values of glucose and calcium were significantly related to those items fore-named. In conclusion, the function of Se-enriched malt in deadening the lesion and delaying the development of hepatoma of rats induced by diethylnitrosamine was better than that of sodium selenite. Hypoglycemia and hypercalcemia were significantly correlated with the multifactors mentioned above. PMID- 16266727 TI - Sublingual administration of Delta9-tetrahydrocannabinol/beta-cyclodextrin complex increases the bioavailability of Delta9-tetrahydrocannabinol in rabbits. AB - The bioavailability of Delta(9)-tetrahydrocannabinol (THC) was determined after its sublingual administration as solid THC/beta-cyclodextrin (THC/beta-CD) complex, and was compared to oral administration of ethanolic THC, in rabbits. The absolute bioavailability of THC after sublingual administration of solid THC/beta-CD complex powder (16.0 +/- 7.5%; mean +/- SD; n = 4) is higher than the bioavailability of THC after oral administration of ethanolic THC solution (1.3 +/- 1.4%; mean +/- SD; n = 4). The results suggest that sublingual administration of THC/beta-CD complex is a useful tool in improving absolute bioavailability of THC. PMID- 16266728 TI - Validation of soft bottom benthic habitats identified by single-beam acoustics. AB - Acoustic diversity charts were produced for a Portuguese soft bottom mid-shelf area, depth from 30 to 90 m, using a single-beam echo sounder coupled to the acoustic systems QTC VIEW Series IV and V. A similar acoustic pattern was identified by both systems, which, after ground-truth interpretation based in available sediment and biological data, established a preliminary spatial distribution model of the benthic habitats in this coastal area. However, some of the acoustic areas were interpreted using one or very few sediment and benthic samples. A specific validation survey was conducted a posteriori, in which the positioning of the sediment and benthic community sampling sites was based on the acoustic diversity previously identified. The results clearly confirm the benthic habitats distribution model suggested by the acoustic method, indicating a high potential for the use of such approach in the identification and mapping of large scale soft bottom coastal shelf habitat diversity. PMID- 16266729 TI - Life-history- and ecosystem-driven variation in composition and residence pattern of seabream species (Perciformes: Sparidae) in two Mediterranean coastal lagoons. AB - Species composition and length-frequency distributions of six sparid fish species were investigated in two central Mediterranean coastal lagoons off the western coast of Italy: Fogliano and Caprolace. In the former, the sparid fauna was dominated by the gilt-head seabream (Sparus aurata), whereas in Caprolace, species composition was more homogeneous across all six species. Size structure varied considerably among species: S. aurata, Diplodus puntazzo and Diplodus vulgaris had a single-cohort structure in both lagoons, whereas in Diplodus annularis and Diplodus sargus at least two cohorts were identified. In Lithognathus mormyrus inter-lagoon variation was detected, with a single-cohort structure in Fogliano and a two-cohort structure in Caprolace. While inter specific differences can be explained by variation in life-history strategies among species, intra-specific variation in L. mormyrus is likely to be determined by the known differences between the two habitats: Fogliano being a more confined lagoon, and Caprolace more extensively influenced by the sea. PMID- 16266730 TI - Aggregate settling velocity of combined sewage overflow. AB - In order to better model the fate and transport of material in a combined sewer overflow (CSO), hydrologic and geochemical measurements were made during a storm event in Flushing Bay, NY, USA. Particle size, total suspended solids concentrations, and CSO solid density are used to calculate the settling velocity of aggregates of the CSO material. Roughly half of the material that is discharged from the outfall sinks rapidly to the bottom, while the other half forms a turbid surface plume in fresher water. The dry solid density of the CSO material was 1589 kg m(-3) and the volume fraction of solid material to total aggregate volume ranged from 0.1 to 1.0. The settling velocity estimates of aggregated material in the surface plume ranged from about 0.5 to 9.0 mm s(-1) with a median settling velocity of 2.9 mm s(-1). A strong inverse relationship between the size of the aggregates and the aggregate density resulted in a fractal dimension of the aggregates of 2.3. PMID- 16266732 TI - Temporal and spatial variation of trace metals in clams Tivela mactroidea along the Venezuelan coast. PMID- 16266731 TI - Anthropogenic iodine-129 in the Arctic Ocean and Nordic Seas: numerical modeling and prognoses. AB - A numerical model simulation has been used to predict extent and variability in the anthropogenic (129)I pollution in the Arctic Ocean and Nordic Seas region over a period of 100 years. The source function of (129)I used in the model is represented by a well-known history of discharges from the Sellafield and La Hague nuclear reprocessing facilities. The simulations suggest a fast transport and large inventory of the anthropogenic (129)I in the Arctic and North Atlantic Oceans. In a fictitious case of abrupt stop of the discharges, a rapid decline of inventories is observed in all compartments except the North Atlantic Ocean, the deep Nordic Seas and the deep Arctic Ocean. Within 15 years after the stop of releases, the model prediction indicates that near-equilibrium conditions are reached in all compartments. PMID- 16266733 TI - Development of a behavioral task measuring reward "wanting" and "liking" in rats. AB - It has been suggested that reward "wanting" and "liking" are mediated by separable brain systems. To facilitate neuropharmacological and neurophysiological research on this issue we developed a behavioral task with putative measures of reward "wanting" and "liking" available on a trial-by-trial basis. We were able to test whether our measures were sensitive to changes in thirsty rats' "wanting" and "liking" of liquid reward by manipulating its delay, taste and volume. We found that three of our putative "wanting" measures (anticipatory errors, reaction time and reward collection latency) were affected by upcoming reward delay and/or taste and our putative "liking" measure (post reward licking) was sensitive to variations in reward taste and volume. To cross validate our measures with previous pharmacological work we tested rats following acute, systemic administration of drug compounds that globally enhance serotonin and noradrenaline (imipramine), dopamine (GBR 12909) and opioid (morphine) function. Imipramine augmented the effects of delay and taste on reward "wanting", GBR 12909 attenuated the effects of delay on reward "wanting" and the effects of taste on reward "liking", and morphine reduced the effect of delay on a measure of reward "wanting". Since morphine failed to affect reward "liking" but has been previously found to enhance reward "liking" in taste reactivity tests, our measure requires further pharmacological validation. However, this task shows potential to assess the specific neural mechanisms that contribute to the impact of reward parameters on "wanting" and "liking". PMID- 16266734 TI - Camalexin induces detoxification of the phytoalexin brassinin in the plant pathogen Leptosphaeria maculans. AB - The impact of the phytoalexins camalexin and spirobrassinin on brassinin detoxification by Leptosphaeria maculans (Desm.) Ces. et de Not. [asexual stage Phoma lingam (Tode ex Fr.) Desm.], a pathogenic fungus prevalent on crucifers, was investigated. Brassinin is a plant metabolite of great significance due to its dual role both as an effective phytoalexin and as an early biosynthetic precursor of the majority of the phytoalexins produced by plants of the family Brassicaceae (Cruciferae). The rate of detoxification of brassinin in cultures of L. maculans increased substantially in the presence of camalexin, whereas spirobrassinin did not appear to have a detectable effect. In addition, the brassinin detoxifying activity of cell-free extracts obtained from cultures incubated with camalexin was substantially higher than that of control cell-free extracts or cultures incubated with spirobrassinin, and correlated positively with brassinin oxidase activity. The discovery of a potent synthetic modulator of brassinin oxidase activity, 3-phenylindole, and comparison with the commercial fungicide thiabendazole is also reported. The overall results indicate that brassinin oxidase production is induced by camalexin and 3-phenylindole but not by spirobrassinin or thiabendazole. Importantly, our work suggests that introduction of the camalexin pathway into plants that produce brassinin might make these plants more susceptible to L. maculans. PMID- 16266735 TI - Early selection of diagnostic facial information in the human visual cortex. AB - There is behavioral evidence that different visual categorization tasks on various types of stimuli (e.g., faces) are sensitive to distinct visual characteristics of the same image, for example, spatial frequencies. However, it has been more difficult to address the question of how early in the processing stream this sensitivity to the information relevant to the categorization task emerges. The current study uses scalp event-related potentials recorded in humans to examine how and when information diagnostic to a particular task is processed during that task versus during a task for which it is not diagnostic. Subjects were shown diagnostic and anti-diagnostic face images for both expression and gender decisions (created using Gosselin and Schyns' Bubbles technique), and asked to perform both tasks on all stimuli. Behaviorally, there was a larger advantage of diagnostic over anti-diagnostic facial images when images designed to be diagnostic for a particular task were shown when performing that task, as compared to performing the other task. Most importantly, this interaction was seen in the amplitude of the occipito-temporal N170, a visual component reflecting a perceptual stage of processing associated with the categorization of faces. When participants performed the gender categorization task, the N170 amplitude was larger when they were presented with gender diagnostic images than with expression-diagnostic images, relative to their respective non-diagnostic stimuli. However, categorizing faces according to their facial expression was not significantly associated with a larger N170 when subjects categorized expression diagnostic cues relative to gender-diagnostic cues. These results show that the influence of higher-level task-oriented processing may take place at the level of visual categorization stages for faces, at least for processes relying on shared diagnostic features with facial identity judgments, such as gender cues. PMID- 16266736 TI - The shape of the anterior and posterior surface of the aging human cornea. AB - PURPOSE: To determine the shape and astigmatism of the posterior corneal surface in a healthy population with age, using Scheimpflug photography corrected for distortion due to the geometry of the Scheimpflug imaging system and the refraction of the anterior corneal surface. METHODS: Scheimpflug imaging was used to measure in six meridians the cornea of the right eye of 114 subjects, ranging in age from 18 to 65 years. RESULTS: The average radius of the anterior corneal surface was 7.79+/-0.27 (SD) mm and the average radius of the posterior corneal surface was 6.53+/-0.25 (SD) mm. Both surfaces were found to be flatter horizontally than vertically. The cylindrical component of the posterior surface of 0.33 mm is twice that of the anterior surface (0.16 mm). The asphericity of both the anterior and the posterior surface was independent of the radius of curvature at the vertex, refractive error and gender. In contrast with that of the anterior corneal surface, the asphericity of the posterior corneal surface varied significantly between meridians. With age, the asphericity of both the anterior and the posterior corneal surface changes significantly, which results in a slight peripheral thinning of the cornea. CONCLUSION: On average, the astigmatism of the posterior corneal surface (-0.305 D) compensates the astigmatism of the anterior corneal surface (0.99 D) with 31%. The results show that the effective refractive index is 1.329, which is lower than values commonly used. There is no correlation between the asphericity of the anterior and the posterior corneal surface. As a result, the shape of the anterior corneal surface provides no definitive basis for knowing the asphericity of the posterior surface. PMID- 16266737 TI - Biomonitoring of mercury in polluted coastal area using transplanted mussels. AB - The Kastela Bay is heavily polluted with inorganic mercury originated from direct discharges from the chlor-alkali plant, which operated in the period from 1950 to 1990. Even though the plant was closed 15 years ago, elevated levels of total mercury are still evident in surface sediments of the bay. In order to assess the availability of remobilized mercury to marine organisms, cultured mussels (M. galloprovincialis) were transplanted from pristine area to Kastela Bay, in the period from September 2000 to March 2001. Mussel samples were collected for the analysis of THg and MeHg in whole soft tissue, gills and digestive gland. Surface sediments and suspended matter were collected for the analysis of THg. Digestive gland was the target organ for the accumulation of THg, while concentrations of MeHg were similar in all analyzed tissues. The percentage of MeHg in mussel tissues (4-27%) was characteristic for the areas contaminated with inorganic mercury. A significant negative correlation was observed between the THg concentration in the tissues and the percentage of MeHg. Concentrations of THg in mussel tissues, which were decreasing from the source of contamination in an anticlockwise direction towards the exit of the bay, were significantly positively correlated to THg content in sediment and suspended particles. Spatial distribution of mercury species (THg and MeHg) in different environmental compartments was in accordance with the prevailing circulation in the bay. Data obtained through 6 months of biomonitoring experiment indicated that digestive gland was more sensitive indicator of THg concentrations in the environment than the whole organism or gills. As for MeHg, all tissues were equally suitable as biomonitors of MeHg concentrations in the environment. PMID- 16266738 TI - Spatial distribution of total Hg in urban soils from an Atlantic coastal city (Aveiro, Portugal). AB - The aim of this study was to investigate the levels and the spatial distribution of total Hg concentrations in soils from the urban area of Aveiro (Portugal) in order to assess the impact of industrial activities and identified Hg emission sources in these urban soils. For this purpose, soils were collected in 25 sampling points (at two depths) within the urban perimeter and in places considered representative of the main green areas of the city. A median concentration of 0.091 mg kg(-1) (dry weight) was obtained, regardless the depth. Aveiro was considered a low polluted city in terms of total Hg and no direct effects of emissions of Hg from industrial activities nearby could be detected in these urban soils. Despite of the low values obtained, high variability (range of approximately 0.5 mg kg(-1)) was observed in the results. Such was considered to be related to characteristic features of soils in urban settings and to the behaviour of Hg in the urban environment. PMID- 16266739 TI - Model simulation of meteorology and air quality during the summer PUMA intensive measurement campaign in the UK West Midlands conurbation. AB - The Regional Atmospheric Modeling System (RAMS) and Urban Airshed Model (UAM IV) have been implemented for prediction of air pollutant concentrations within the West Midlands conurbation of the United Kingdom. The modelling results for wind speed, direction and temperature are in reasonable agreement with observations for two stations, one in a rural area and the other in an urban area. Predictions of surface temperature are generally good for both stations, but the results suggest that the quality of temperature prediction is sensitive to whether cloud cover is reproduced reliably by the model. Wind direction is captured very well by the model, while wind speed is generally overestimated. The air pollution climate of the UK West Midlands is very different to those for which the UAM model was primarily developed, and the methods used to overcome these limitations are described. The model shows a tendency towards under-prediction of primary pollutant (NOx and CO) concentrations, but with suitable attention to boundary conditions and vertical profiles gives fairly good predictions of ozone concentrations. Hourly updating of chemical concentration boundary conditions yields the best results, with input of vertical profiles desirable. The model seriously underpredicts NO2/NO ratios within the urban area and this appears to relate to inadequate production of peroxy radicals. Overall, the chemical reactivity predicted by the model appears to fall well below that occurring in the atmosphere. PMID- 16266740 TI - Cadmium and lead contamination in japonica rice grains and its variation among the different locations in southeast China. AB - On the basis of a large-scale survey with 269 sampling sites and a field experiment with 12 cultivars grown at 3 ecologically different locations, Cadmium (Cd) and lead (Pb) contamination in milled rice grains and its variation among different locations were investigated in the area of southeast China. The objective of this study was mainly to assess the present situation of Cd and Pb contamination in rice grains harvested locally and find out the variation of Cd and Pb background levels in rice grains among the diverse regional locations. The results showed that: a) Cd concentrations in milled rice grains ranged from trace (below 10 ng/g) to 340 ng/g, with the means of 81.4 ng/g (AM) and 55.2 ng/g (GM) for the 269 samples, whereas Pb concentrations ranged from trace (below 10 ng/g) to 1136 ng/g, with the means of 113.5 ng/g (AM) and 51.0 ng/g (GM), respectively. High frequency was observed for both the Cd and Pb concentrations ranging from trace to 200 ng/g, with 95.9% for Cd and 84.8% for Pb, respectively. b) ANOVA for the AMs and GMs among 269 sampling sites indicated that there were significant differences among the different spatial locations in Cd or Pb concentrations. c) The Cd concentrations in milled rice grains were poorly correlated with Pb concentration among the 269 sampling sites surveyed, and no significant difference (P>0.05) in the AMs or GMs of Cd and Pb concentration was observed among different cities within a range of the same provinces, only with an exception for 1-2 provinces. d) Variance analysis of Cd, Pb concentrations for 12 rice cultivars grown in three locations showed that the great difference in coefficients of variation (CV) was observed both among cultivars grown in the same location and among locations of cultivars planted, in which locations showed much more contribution to the variation of Cd or Pb concentration in milled rice grains as compared with cultivars. PMID- 16266741 TI - Mercury distribution in fish organs and food regimes: Significant relationships from twelve species collected in French Guiana (Amazonian basin). AB - Within a multidisciplinary research programme set up in French Guiana (Amazonian basin), twelve fish species from six food regimes were collected from the upper part of the Maroni River in order to analyze mercury (Hg) distribution in six organs (gills, liver, kidneys, skeletal muscle, stomach, and intestine) and to look for a relationship between Hg organotropism and food regimes. As many studies have shown, mercury biomagnification leads to extremely marked differences in muscle accumulation levels: the average ratio between extreme concentrations measured in piscivorous and herbivorous species was almost 500. A first principal component analysis on primary Hg concentration variables showed that biomagnification had a marked effect, masking differences between Hg distribution in the organs according to fish species and their food regimes. In order to avoid this, we determined ratios between Hg concentrations measured in the different organs and in the skeletal muscle, considered as the reference tissue for biomagnification effects. A new principal component analysis using these normalized values, in conjunction with a Ward's hierarchical clustering method, revealed that there is a link between Hg organotropism and the food regimes, with comparatively high [Hg]gills/[Hg]muscle ratios for the herbivorous species; high [Hg]intestine-liver-kidneys/[Hg]muscle ratios for the benthivorous and periphytophagous species, and, in contrast, ratios of less than 1 in the different organs for the piscivorous and omnivorous species. Our determinations of methylmercury (MMHg) percentages in the food consumed by the fish (aquatic macrophytes, terrestrial material from the river banks, biofilms, benthic invertebrates, fish muscle tissues), according to the different food regimes (herbivorous, periphytophagous, benthivorous, omnivorous, carnivorous, piscivorous), showed that this criterion can account for the differences in Hg distribution in the fish organs. For instance, the periphytophagous and benthivorous fish species ingest biofilms and small benthic invertebrates with quite low MMHg burdens (18% and 35 to 52% of Hgtotal, respectively). The highest [Hg]organs/[Hg]muscle ratios were observed for the liver and kidneys, the two principal target organs for inorganic Hg in fish. On the other hand, the piscivorous species ingest a large amount of fish of varying size, with high MMHg percentages in their muscle tissue (nearly 80%); Hg organotropism is characterized by high MMHg concentrations in the skeletal muscle and comparatively low [Hg]organs/[Hg]muscle ratios. PMID- 16266742 TI - Shorter PFA-100-closure times in laparoscopic versus conventional hysterectomy are not caused by increased vasopressin levels. AB - BACKGROUND/OBJECTIVES: Bleeding problems during laparoscopic surgery are infrequent. We hypothesised that increased abdominal pressure during the application of the pneumoperitoneum would lead to an increased release of endogenous vasopressin which could then contribute to the hemostasis by increasing platelet reactivity, FVIII and von Willebrand-factor. PATIENTS AND METHODS: We compared the vasopressin levels, the platelet function as measured by the PFA-100-test, aPTT and FVIII in 39 consecutive patients who underwent elective hysterectomy (20 with the laparoscopic and 19 with the conventional, "open" method). Blood was sampled the day before surgery and 2, 4 and 72 h after the induction of anaesthesia. RESULTS: After two hours, the PFA-100 closure times with collagen/ADP decreased to lower levels in the laparoscopic group (from 93 +/ 22 to 82 +/- 20, mean +/- SD) and even further down to 65 +/- 13 s (compared to 82 +/- 20 s) (p = 0.024)) four hours after the beginning of surgery. Vasopressin levels and F VIII increased in both groups but there was no significant difference between the groups (21 vs. 17.8 pmol/l for vasopressin, differences of the mean). Bleeding was minimal, with a trend to lower Hb-levels in the laparotomy group. CONCLUSIONS: The procedural difference of laparoscopic vs. open hysterectomy appears to enhance platelet reactivity by other mechanisms than increased vasopressin levels and may contribute to an enhanced hemostatic competence in laparoscopic surgery. PMID- 16266744 TI - Villoglandular adenocarcinoma of the cervix: clarity is needed on the histological definition for this difficult diagnosis. AB - BACKGROUND: Villoglandular adenocarcinoma (VGA) of the cervix is reported as a variant of a cervical adenocarcinoma with a good prognosis. CASES: We present two cases histologically reported as a villoglandular adenocarcinoma of the cervix that have recurred and progressed rapidly since initial treatment. External histopathological review suggested both had a prominent villoglandular pattern but with an associated underlying well-differentiated adenocarcinoma. CONCLUSION: The diagnosis of VGA is difficult. Current literature is not entirely consistent in the presented definition, and further clarity is needed. Because of the rarity of VGA and the difficulty but importance of the diagnosis, we would feel that a central review of all cases of VGA is warranted. This would assist in diagnosis and also in obtaining accurate follow-up data. PMID- 16266743 TI - Sequential gemcitabine-carboplatin followed by paclitaxel-carboplatin in the first-line treatment of advanced ovarian cancer: A phase II study. AB - OBJECTIVE: To determine the feasibility and efficacy of sequential gemcitabine carboplatin followed by paclitaxel-carboplatin in the first-line treatment of advanced epithelial ovarian cancer, with the response rate as the primary endpoint. METHODS: After primary laparotomy, 56 patients with FIGO Stages III-IV disease were given 4 cycles of gemcitabine 1000 mg/m2 d1,8 and carboplatin AUC5 (44 patients) or AUC6 (12 patients) d1 q3wk followed by 4 cycles of paclitaxel 175 mg/m2 d1 and carboplatin AUC5/6 q3wk. Of the tumors, 43 were serous, 6 clear cell, 4 endometrioid, and 3 anaplastic type. Thirty-seven (66.1%) of the patients were suboptimally debulked. RESULTS: Forty-seven patients were evaluable for response by CA-125 criteria, and 46 (98%) responded. Thirty patients (after gemcitabine-carboplatin) and 24 (after paclitaxel-carboplatin) were evaluable for response by CT (RECIST criteria), respectively. After the four gemcitabine carboplatin cycles, the objective response rate was 83% (6 CR, 19 PR). Following completion of the whole sequential regimen, 7 patients showed a CR and 15 a PR, respectively, giving a response rate of 92%. The median progression-free survival was 12.8 months after a median follow-up of 19 months (range 7-35 months). The median overall survival has not been reached yet. The main toxicity was neutropenia as 139/221 (62.9%) of the gemcitabine-carboplatin cycles and 92/181 (50.8%) of the paclitaxel-carboplatin cycles, respectively, were associated with Grades 3-4 neutropenia. Neutropenia was reported as a serious adverse event in 5.7% of the cycles, and G-CSF support was needed in 18.4% of the cycles. Only the gemcitabine-carboplatin cycles were associated with a marked thrombocytopenia (32.1% Grades 3-4). Of the other side effects, marked allergy occurred in 14/52 (27%) exposed to paclitaxel. A total of 14 patients discontinued the treatment prematurely: 3 due to lack of efficacy, 1 due to protocol violation, and 10 due to toxicity (4 allergic reactions to paclitaxel, 3 complicated neutropenias, 1 fever, and 2 unspecified toxicities). The mean relative dose intensities were: gemcitabine 84.0%, paclitaxel 85.4%, and carboplatin 96.5%. Of the gemcitabine carboplatin cycles and paclitaxel-carboplatin cycles, 32% and 38% were delayed, respectively. Gemcitabine d8 dose had to be omitted in 8% of the cycles. CONCLUSION: The sequential regimen of gemcitabine-carboplatin followed by paclitaxel-carboplatin is feasible in chemotherapy-naive ovarian cancer. Although its use is associated with a marked neutropenia, the neutropenia is manageable. PMID- 16266745 TI - Errors in development of fetuses and placentas from in vitro-produced bovine embryos. AB - In vitro systems for oocyte maturation, fertilization and embryo culture [in vitro production (IVP)] have the potential for more wide-spread use in creative breeding programs for dairy and beef cattle. However, one negative consequence of both IVP and somatic cell nuclear transfer (SCNT) in cattle and other species is that embryos, fetuses, placentas, and offspring can differ significantly in morphology and developmental competence compared with those from embryos produced in vivo. Fetuses and placentas derived from IVP and SCNT embryos may fall within the normal range of development, may have obvious abnormalities such as increased fetal and placental weights, or may have subtle abnormalities such as aberrant development of fetal skeletal muscle, placental blood vessels, and altered metabolism. Failures in physiologic and/or genetic mechanisms essential for proper fetal growth and survival outside of the uterus contribute significantly to pregnancy and neonatal losses. Oversized fetuses are at increased risk of death during parturition and the adverse consequences of severe dystocia may compromise the dam. Collectively, these abnormalities have been referred to as 'large offspring syndrome' or 'large calf syndrome'. Abnormal phenotypes resulting from IVP and SCNT embryos are stochastic in occurrence and they have not been consistently linked to aberrant expression of single genes or specific pathophysiology. Thus, reliable methods of early diagnosis of the condition are not yet available. The objective of this paper is to examine abnormal development of fetuses and placentas resulting from embryos produced using in vitro systems. The term 'abnormal offspring syndrome (AOS)' is introduced and a classification system of developmental outcomes is proposed to facilitate research efforts on the mechanisms of the various abnormal phenotypes. We also discuss potential genetic and physiologic mechanisms that may contribute to abnormal phenotypes following transfer of IVP and SCNT embryos. PMID- 16266746 TI - DNA-collagen complex as a carrier for Ag+ delivery. AB - The possibility of DNA-collagen complex as a drug carrier was investigated. The interaction between DNA and silver ions was proved by CD spectra. The release property of the complex of DNA-Ag+ was measured through turbidity of PBS solution to indicate that silver ions could coordinate with base pairs of DNA, and be released slowly from the complex of DNA-Ag+. Collagen film, collagen-Ag+ film, DNA-collagen film and DNA-collagen-Ag+ film were prepared, and studied through SEM. Particles were found present in DNA-collagen-Ag+ film by SEM. These show that silver ions may be enclosed inside these particles, which led to the slow release of Ag+ to the environments. Two bacteria, Escherichia coli and Staphylococcus aureus, were used to study the antibiotic properties of the complex films. The growth of E. coli and S. aureus could be inhibited by these films. It indicates that DNA-collagen may be a good drug carrier for the drug controlled release. PMID- 16266747 TI - Fasting-induced increases of arcuate NPY mRNA and plasma corticosterone are blunted in the rat experienced neonatal maternal separation. AB - This study was conducted to examine the effects of neonatal maternal separation on the hypothalamic expression of feeding peptides in later life. Pups in maternal separation (MS) groups were separated from their dam for 3 h daily from postnatal day (PND) 1-14, while pups in non-handled (NH) groups were left undisturbed. Rats were sacrificed on PND 60 to examine the gene expression of neuropeptide Y (NPY) and pro-opiomelanocortin (POMC) in the hypothalamic arcuate nucleus by mRNA in situ hybridization. Half of the rats from each group were food deprived for 48 h before sacrifice. POMC mRNA expression increased in the free fed MS group compared with the free fed NH group. Food deprivation significantly decreased the arcuate POMC mRNA level in both groups. Body weight gain, basal levels of plasma corticosterone, leptin, and arcuate NPY mRNA were not modulated by experience of neonatal maternal separation. However, fasting-induced increases of plasma corticosterone and arcuate NPY expression were blunted in MS rats. These results suggest that neonatal maternal separation may increase the basal expression level of arcuate POMC mRNA, while inhibit the fasting-induced expression of arcuate NPY mRNA, later in life. Lastly, the altered expression of arcuate NPY mRNA, but not of arcuate POMC mRNA, appeared to be related with altered activity of the hypothalamic-pituitary-adrenal gland axis in offspring by neonatal maternal separation. PMID- 16266748 TI - Prevalence of mental disorders and deliberate self-harm in Greek male prisoners. AB - The aim of this survey was to determine the prevalence of current and lifetime mental disorder and deliberate self-harm among male prisoners in Greece. The subjects were 80 randomly selected remanded and sentenced prisoners in a Greek prison. They were assessed for mental disorder including suicidality and substance misuse using the Mini International Neuropsychiatric Interview (MINI). We also collected information regarding contact with psychiatric services, previous deliberate self-harm as well as physical health and conducted a brief assessment of their intellectual functioning. Mental disorder was diagnosed in 63 (78.7%) prisoners. The main diagnoses were: anxiety disorder, 30 (37.5%); major depression, 22 (27.5%); antisocial personality disorder, 30 (37.5%); alcohol dependence, 21 (26.3%) and opiate dependence 22 (27.5%) and schizophrenic or bipolar disorder 9 (11.2%). Deliberate self-harm prior to and during imprisonment was reported by 15% and 2.5% of prisoners, respectively, and 12 prisoners (15%) had IQ below 75. This survey identified a significant level of need for specialist mental health services in prison. Further studies are required to assess the specific needs of those patients who are too unwell to remain in prison, the need for specific treatments for substance misuse and improved assessment/treatment of common psychiatric disorders. PMID- 16266749 TI - Collagen type I-mediated activation of ERK/MAP Kinase is dependent on Ras, Raf-1 and protein phosphatase 2A in Jurkat T cells. AB - Growing evidence indicates that interactions of T cells with extracellular matrix through beta1 integrins are important for the regulation of T cell-mediated immune responses and diseases. In this regard, we have recently demonstrated that collagen I (Coll I) through alpha2beta1 integrin inhibited Fas-induced apoptosis of T cells by activating a protein phosphatase 2A (PP2A)-dependent ERK/MAP Kinase pathway. As survival of T cells is critical for their functions, we further investigated the mechanisms underlying the activation of this pathway. Inhibition studies demonstrated that Coll I activates the ERK/MAP Kinase pathway in Jurkat T cells through the activation of Ras and Raf-1. Activation of PP2A was not necessary for the binding of Coll I to Jurkat T cells, but is required for the activation of Raf-1. In accordance, activation of Ras, Raf-1 and PP2A were also required for the ability of Coll I to protect Jurkat T cells from Fas-induced apoptosis. In contrast and despite its capacity to activate Ras, fibronectin (Fbn) failed to activate PP2A and Raf-1. These results might explain, at least in part, the weak ability of Fbn to activate ERK in T cells, supporting thus the differential signaling of beta1 integrin members in these cells. This study provides novel insights into the mechanisms by which beta1 integrins activate the ERK/MAP Kinase pathway in T cells, and is the first report to provide a role for PP2A in integrin-mediated ERK/MAP Kinase activation. PMID- 16266750 TI - Effects of cadmium on Na+ transport in the isolated skin of the toad Pleurodema thaul. AB - Cadmium ions applied to either (outer or inner) surface of the isolated toad skin dose-dependently increased the short-circuit current (SCC), the potential difference (V) and the active sodium conductance (G(Na)) in the concentration range 0.07-0.50mM. Maximal stimulatory effect was over 30% with an EC(50) of about 0.1mM. The effect of the highest concentration used (0.75mM) decreased considerably, and when it was applied to the inner surface (10 experiments), induced between 30% and 40% inhibition of the electric parameters in four experiments. Pretreatment with amiloride inverted the stimulatory effect of externally applied Cd(2+), suggesting competitive action on the apical Na(+) channel. The effect of noradrenaline (NA) was increased after outer application of Cd(2+) and decreased after inner application of the metal: the latter effect might be due to cadmium inhibition of the activity of Na(+),K(+)-ATPase. On the other hand, pretreatment with amiloride was followed by partial although transient reversal of its effects by serosal Cd(2+), which might be explained by action of cadmium on cytoplasmic lysine residues concerned with Na(+) channel gating. The amiloride test showed that the increment of the electric parameters was due principally to stimulation of the driving potential for Na(+) (V E(Na(+))) and that inhibition was accompanied by a reduction in the V-E(Na(+)) and by a significant decrease in skin resistance indicating possible disruption of membrane or cell integrity. These data are in favor of the possibility that externally applied Cd(2+) activates toad skin ion transport, partly by increasing apical sodium conductance and also by stimulating the V-E(Na(+)), and that internally applied Cd(2+), with easier access to membrane and cellular constituents, may inhibit the sodium pump. PMID- 16266751 TI - Hg(II)-coordination by sugar-acids: role of the hydroxy groups. AB - A solution study on the ability of some derivatised sugars [glucuronic acid (GluA), galacturonic acid (GalA) and glucosaminic acid (GlNA)] to complex the Hg(II) ion is reported. The stability constants of the complex species were determined by potentiometric measurements while (1)H NMR experiments allow to define the coordination sites of sugar molecules. GluA coordinates the metal ion through the carboxylic oxygen and the O-4 hydroxyl group and is found to form more stable complexes with respect to GalA in which metal ligation is from the carboxylic oxygen and the O-5 ring oxygen. GlNA forms stable complexes chelating Hg(II) ion through carboxylic oxygen and the alpha-amino group. The ternary 2,2' bipyridine containing systems were also investigated by means of potentiometric studies. The ML(2) complexes were also isolated in the solid state and characterised by IR spectroscopy. PMID- 16266752 TI - Mechanism of action of sphingolipids and their metabolites in the toxicity of fumonisin B1. AB - Fumonisins are a group of mycotoxins produced primarily by Fusarium moniliforme. Several fumonisins have been isolated through out the years but only fumonisin B1, B2 and B3 are the ones present in naturally contaminated foods, with B1 being the most toxic between them. The structural similarity between sphinganine and fumonisin B1 suggests that the mechanism of action of this mycotoxin is mainly via disruption of sphingolipid metabolism, this is an important step in the cascade of events leading to altered cell growth, differentiation and cell injury. Sphingolipids are a second type of lipid found in cell membranes, particularly nerve cells and brain tissues. Toxicity of fumonisin B1 is given via inhibition of ceramide synthase that catalyzes the formation of dihydroceramide from sphingosine. This mechanism of action may explain the wide variety of health effects observed when this mycotoxin is ingested like high rate of human oesophageal cancer and promotion of primary liver cancer. PMID- 16266753 TI - Symptoms following abrupt discontinuation of duloxetine treatment in patients with major depressive disorder. AB - BACKGROUND: Discontinuation symptoms are common following antidepressant treatment. This report characterizes symptoms following duloxetine discontinuation. METHODS: Data were obtained from 9 clinical trials assessing the efficacy and safety of duloxetine in the treatment of major depressive disorder (MDD). RESULTS: In a pooled analysis of 6 short-term treatment trials, in which treatment was stopped abruptly, discontinuation-emergent adverse events (DEAEs) were reported by 44.3% and 22.9% of duloxetine- and placebo-treated patients, respectively (p<0.05). Among duloxetine-treated patients reporting at least 1 DEAE, the mean number of symptoms was 2.4. DEAEs reported significantly more frequently on abrupt discontinuation of duloxetine compared with placebo were dizziness (12.4%), nausea (5.9%), headache (5.3%), paresthesia (2.9%), vomiting (2.4%), irritability (2.4%), and nightmares (2.0%). Dizziness was also the most frequently reported DEAE in the analyses of 3 long-term duloxetine studies. Across the short- and long-term data sets, 45.1% of DEAEs had resolved in the duloxetine-treated populations by the end of the respective studies, and the majority of these (65.0%) resolved within 7 days. Most patients rated the severity of their symptoms as mild or moderate. A higher proportion of patients reporting DEAEs were seen with 120 mg/day duloxetine compared with lower doses. For doses between 40 and 120 mg/day duloxetine the proportion of patients reporting at least one DEAE differed significantly from placebo. Extended treatment with duloxetine beyond 8-9 weeks did not appear to be associated with an increased incidence or severity of DEAEs. CONCLUSIONS: Abrupt discontinuation of duloxetine is associated with a DEAE profile similar to that seen with other selective serotonin reuptake inhibitor (SSRI) and selective serotonin and norepinephrine reuptake inhibitor (SNRI) antidepressants. It is recommended that, whenever possible, clinicians gradually reduce the dose no less than 2 weeks before discontinuation of duloxetine treatment. LIMITATIONS: The main limitation is the use of spontaneously reported DEAEs. PMID- 16266754 TI - Lifespan development of pro- and anti-saccades: multiple regression models for point estimates. AB - The comparative study of anti- and pro-saccade task performance contributes to our functional understanding of the frontal lobes, their alterations in psychiatric or neurological populations, and their changes during the life span. In the present study, we apply regression analysis to model life span developmental effects on various pro- and anti-saccade task parameters, using data of a non-representative sample of 327 participants aged 9 to 88 years. Development up to the age of about 27 years was dominated by curvilinear rather than linear effects of age. Furthermore, the largest developmental differences were found for intra-subject variability measures and the anti-saccade task parameters. Ageing, by contrast, had the shape of a global linear decline of the investigated saccade functions, lacking the differential effects of age observed during development. While these results do support the assumption that frontal lobe functions can be distinguished from other functions by their strong and protracted development, they do not confirm the assumption of disproportionate deterioration of frontal lobe functions with ageing. We finally show that the regression models applied here to quantify life span developmental effects can also be used for individual predictions in applied research contexts or clinical practice. PMID- 16266755 TI - Real-time reverse transcription polymerase chain reaction assay to detect a broad range of feline calicivirus isolates. AB - This report describes a real-time reverse transcription polymerase chain reaction (RT-PCR) assay with SYBR Green targeting the VP2 (ORF 3) of feline caliciviruses. All of the 44 feline calicivirus isolates tested were detected, whereas neither feline herpesvirus, feline panleukopenia virus, feline immunodeficiency virus and feline leukemia virus nor other calicivirus like rabbit hemorrhagic disease virus and a canine calicivirus isolate showed specific amplification products. The sensitivity of the SYBR Green reaction was shown to be equivalent to 5 x 10(1) to 5 x 10(2) copies/reaction and the overall sensitivity equivalent to a feline calicivirus titer of 10(0.6) TCID(50)/100 microl in Crandell Reese Feline Kidney (CRFK) cells. PMID- 16266756 TI - Purification of adenoviral vectors using expanded bed chromatography. AB - The increasing numbers of pre-clinical and clinical trials where recombinant adenoviral vectors are used for gene therapy and vaccination require the development of cost-effective and reproducible large scale purification strategies of the biologically active particles. Alternatives to the traditional laboratory scale CsCl density gradient ultracentrifugation method, such as fixed bed chromatography strategies, have been developed, but the yields of final recovery remain too low due mainly to the capture and concentration steps taking place before and between the chromatographic stages. In this study, a rapid and efficient scale-able purification protocol allowing to obtain concentrated, pure and bioactive adenoviral vectors was developed. This allows efficient levels of binding to the column media and vector purification without centrifugation or filtration steps. Expanded bed chromatography followed by hollow fiber concentration allows the capture of viral particles directly from cellular extracts with high efficiency and vector purification is achieved in less than one working day with a minimal amount of sample handling, thus presenting an improvement over existing processes. The overall process yield reached 32%, representing an eight-fold improvement over results reported previously, while the purity is comparable to that obtained with the CsCl method. PMID- 16266757 TI - Golgi and centrosome cycles in Toxoplasma gondii. PMID- 16266758 TI - Propagation velocity, contractility and prognosis in patients with delayed relaxation. PMID- 16266759 TI - Association between post-ischemic forearm blood flow and blood pressure response to maximal exercise in well trained healthy young men. AB - BACKGROUND: The relationship between peripheral circulation and blood pressure (BP) response to maximal exercise is an intriguing and not yet well defined topic. AIMS: Aims of the present study were to investigate in well trained young healthy males the possible relationships between the endothelial or the smooth muscle component of the peripheral circulation and 1) the BP response to physical exercise on treadmill 2) the body mass composition. SUBJECTS AND METHODS: Fifteen subjects (18-36 years), regularly performing physical activity 3 times weekly underwent the following examinations: body composition by bioelectrical impedance analysis; measurement of the forearm blood flow (FBF) at rest and during post ischemic hyperemia by strain-gauge plethysmography at the upper arm; measurement of brachial artery diameter (BAD) at rest and after 4-min ischemia by echography; BP response to maximal exercise on treadmill with the determination of maximal oxygen consumption and the measurement of lactic acid serum concentration. RESULTS: BAD was significantly increased during post-ischemic hyperemia up to the 4th minute of observation with a peak at 60 s (+8.5%); FBF increased at 30 s after ischemia (+210%) and returned to baseline levels at the 2nd minute. In the linear correlation analysis, systolic BP increase at the end of the maximal exercise was significantly and inversely related to the increase in FBF (r= 0.663, p<0.01) and to the early FMD (r=0.503, p<0.05). In the multiple regression analysis, however, only FBF independently affected SBP increase during exercise (t=-3.268, p<0.02). Systolic BP increase at the end of the maximal exercise was significantly related to the increase in FBF but not to that of BAD. Among parameters of body composition, fat-free mass was closely related to changes only in BAD. CONCLUSIONS: These data indicate that FBF, which depends on the smooth muscle component of the peripheral circulation, is closely related to BP response to exercise while the endothelial function, which has been determined as changes in BAD, is related to the fat-free mass of the body, possibly through the peripheral insulin sensitivity. PMID- 16266760 TI - Optimizing heart failure management: an Australian experience. PMID- 16266761 TI - Prior clopidogrel use and outcome of acute coronary syndrome. AB - BACKGROUND: Prior aspirin use has been associated with poorer outcome in acute coronary syndrome, and forms part of the TIMI Risk Score. It is not known if prior use of clopidogrel is associated with similar risk. AIM: To assess if prior clopidogrel use is associated with higher risk in acute coronary syndrome. PARTICIPANTS: Participants were 869 consecutive admissions to a Scottish district general hospital with suspected acute coronary syndrome. METHODS: Incidence of death, recurrent myocardial infarction or urgent percutaneous intervention at 2 weeks was recorded. Odds ratios for sub-groups on clopidogrel, aspirin or neither were calculated. RESULTS: Odds ratios were: clopidogrel 1.46 (95% ci 0.62-3.33), aspirin 1.09 (95% ci 0.64-1.85), neither 0.91 (95% ci 0.53-1.54). CONCLUSION: No definite association was shown between clopidogrel use and outcome but there was a trend towards increased risk of major acute coronary events. PMID- 16266762 TI - Novel mitochondrial DNA mutations in a rare variety of hypertrophic cardiomyopathy. AB - We report a rare case of a 65 year old male with mid left ventricular cavity obstruction which is an uncommon form of hypertrophic cardiomyopathy with cytogenetic analysis revealing novel mutations in mitochondrial nucleic acid. PMID- 16266763 TI - Animal health pharmaceutical industry. AB - The animal health pharmaceutical industry has proactively reported on the volumes of member company antimicrobial active ingredients sold in the U.S. At the individual company level, reporting of finished product distribution data to the FDA is a regulatory requirement, with applications to surveillance and pharmacovigilance. An accounting of product manufactured is done for purposes of good business practices, as well as marketing analyses. Additional applications of antimicrobial usage data might include use in risk assessments, such as for the FDA's Center for Veterinary Medicine Guidance for Industry #152 for the evaluation of the microbiological safety of antimicrobials intended for use in food animals. Compilation of national usage data will be a complex undertaking, hindered by issues such as confidentiality, auditing, field use practice variations, population dynamics (e.g. disease incidence, market conditions for poultry and livestock production), and generic usage. The amounts or volumes in pounds should be considered relative to the large number of animals under husbandry in the United States. Large volumes might seem impressive unless put into proper context. Until such time as a clearly defined application of national usage data is agreed, it is recommended that local usage programs will provide more useful information to perpetuate prudent antimicrobial use in animals. PMID- 16266764 TI - Stakeholder position paper: the need for antimicrobial use data for risk assessment. AB - Risk assessment seeks to estimate the probability and impact of human health effects due to antimicrobial resistance arising from antimicrobial use in animals. Potential ecological pathways for the flow of antimicrobial resistant bacteria and resistance determinants through the food supply and environment are diverse and complicated and this has been a logistical barrier to direct epidemiological measure of risk. Consequently, a number of indirect approaches to assessment have been developed. This paper provides some examples of risk assessments that have been conducted with and without antimicrobial use data, and identifies possible benefits and applications of quantitative antimicrobial use data for risk assessment. PMID- 16266765 TI - Rapid, specific and quantitative assays for the detection of the endophytic bacterium Methylobacterium mesophilicum in plants. AB - Xylella fastidiosa is a xylem-limited bacterium that causes citrus variegated chlorosis disease in sweet orange. There is evidence that X. fastidiosa interacts with endophytic bacteria present in the xylem of sweet orange, and that these interactions, particularly with Methylobacterium mesophilicum, may affect disease progress. However, these interactions cannot be evaluated in detail until efficient methods for detection and enumeration of these bacteria in planta are developed. We have previously developed standard and quantitative PCR-based assays specific for X. fastidiosa using the LightCycler system [Li, W.B., Pria Jr., L.P.M.W.D., X. Qin, and J.S. Hartung, 2003. Presence of Xylella fastidiosa in sweet orange fruit and seeds and its transmission to seedlings. Phytopathology 93:953-958.], and now report the development of both standard and quantitative PCR assays for M. mesophilicum. The assays are specific for M. mesophilicum and do not amplify DNA from other species of Methylobacterium or other bacteria commonly associated with citrus or plant tissue. Other bacteria tested included Curtobacterium flaccumfaciens, Pantoea agglomerans, Enterobacter cloacae, Bacillus sp., X. fastidiosa, Xanthomonas axonopodis pv. citri, and Candidatus Liberibacter asiaticus. We have demonstrated that with these methods we can quantitatively monitor the colonization of xylem by M. mesophilicum during the course of disease development in plants artificially inoculated with both bacteria. PMID- 16266766 TI - Inhibitory effects of microencapsulated allyl isothiocyanate (AIT) against Escherichia coli O157:H7 in refrigerated, nitrogen packed, finely chopped beef. AB - Allyl isothiocyanate (AIT) is an effective inhibitor of various pathogens, but its use in the food industry is limited by its volatility and pungency. The objective of this study was to overcome the volatility of AIT by microencapsulation and evaluate its antimicrobial effectiveness against Escherichia coli O157:H7 in chopped beef. Chopped beef was aseptically prepared and inoculated with a five-strain cocktail of E. coli O157:H7 to yield 4 or 8 log10 cfu/g. AIT was microencapsulated in gum acacia to yield 3.7-54.8 mg AIT/g at a ratio of 1:4 and freeze dried. Microcapsules at 5% or 10% (w/w) were then added to experimental samples that were packed under nitrogen, and stored at 4 degrees C for 18 days. Samples were analyzed for numbers of E. coli O157:H7 and the aerobic mesophilic bacteria (TAC) at 3-day intervals. AIT at 4980 ppm eliminated both low and high levels of inoculated E. coli O157:H7 after 15 and 18 days of storage, respectively. AIT at 2828 ppm reduced E. coli by 2.7 log10 cfu/g by 18 days of storage. AIT levels <1000 ppm were not more effective in reducing E. coli survival than the control treatment without AIT addition. AIT at 170-1480 ppm had negligible effects on the TAC, and while 4980 ppm kept TAC levels 80% were prepared using the double emulsion method. PEG-PH significantly improved the stability of BSA both in aqueous solutions and in PLGA microspheres. The release profiles of BSA from different formulations of PLGA microspheres were significantly different. PEG-PH effectively buffered the local acidity inside the microspheres and improved BSA release kinetics by reducing initial burst release and extending continuous release over a period of time, when encapsulated as an ionic complex. PLGA degradation rate was found to be delayed by PEG-PH. There was clear evidence that PEG-PH played multiple roles when complexed with BSA and incorporated into PLGA microspheres. PEG-PH is an effective excipient for preserving the structural stability of BSA in aqueous solution and BSA/PLGA microspheres formulation. PMID- 16266770 TI - Genes required for fructose metabolism are expressed in Purkinje cells in the cerebellum. AB - Since 1967, fructose has become the primary commercial sweetener in the food industry. Large amounts of fructose can be toxic and have been correlated with atherosclerosis, malabsorption, hyperuricemia, lactic acidosis, and cataracts. To understand the deleterious and critical role(s) fructose plays in normal metabolism, it is essential to know how and where fructose is metabolized. The fructose transporter, GLUT5, and the specialized enzymes ketohexokinase, aldolase, and triokinase comprise the well-defined fructose-specific metabolic pathway found in liver, kidney, and small intestine. It is estimated that 50-70% of ingested fructose is metabolized in these tissues; where and how the remaining 30-50% is metabolized is not well defined. Prediction of tissues capable of metabolizing fructose via this pathway was done using expressed sequence tags (ESTs) in Unigene and a gene-specific virtual northern blot (VNB) algorithm. Unigene and VNB combined correctly predicted the expression of the genes required for fructose metabolism in liver, kidney, and small intestine. Both methods indicated brain, breast, lymphocytes, muscle, placenta, and stomach additionally express this set of genes. Expression of the genes for GLUT5 (glut5) and ketohexokinase (khk) in neurons was validated by immunohistochemistry and RNA in situ hybridization, respectively. Using stringent controls, clear expression of glut5 and khk was localized to Purkinje cells in the cerebellum. Cerebellum was used to oxidize fructose to carbon dioxide. Together, these data suggest that these neurons in the brain are able to utilize fructose as a carbon source. PMID- 16266771 TI - Inhibition of PC5 expression decreases CCK secretion and increases PC2 expression. AB - Cholecystokinin (CCK) is produced from pro CCK by a series of enzymatic cleavages. One of the enzymes thought to be important for pro CCK cleavage is prohormone convertase 5 (PC5). STC-1 cells, a mouse intestinal tumor cell line that expresses CCK, PC1, PC2, and PC5 were stably transfected with hairpin loop plasmids encoding siRNA targeting PC5 and clones were selected. CCK secretion was reduced significantly. PC5 mRNA and protein expression as measured by quantitative PCR and Western blot analysis was reduced about 50%. CCK and PC1 mRNA expression were not changed. These cells showed a three-fold increase in PC2 mRNA and protein expression. This increase may represent a compensatory mechanism triggered by the loss of PC5. The decrease in CCK in the media was due largely to loss of CCK 22. These results provide the first direct evidence that PC5 is involved in CCK processing. PMID- 16266772 TI - Prevention of Alzheimer's disease: Omega-3 fatty acid and phenolic anti-oxidant interventions. AB - Alzheimer's disease (AD) and cardiovascular disease (CVD) are syndromes of aging that share analogous lesions and risk factors, involving lipoproteins, oxidative damage and inflammation. Unlike in CVD, in AD, sensitive biomarkers are unknown, and high-risk groups are understudied. To identify potential prevention strategies in AD, we have focused on pre-clinical models (transgenic and amyloid infusion models), testing dietary/lifestyle factors strongly implicated in reducing risk in epidemiological studies. Initially, we reported the impact of non-steroidal anti-inflammatory drugs (NSAIDs), notably ibuprofen, which reduced amyloid accumulation, but suppressed few inflammatory markers and without reducing oxidative damage. Safety concerns with chronic NSAIDs led to a screen of alternative NSAIDs and identification of the phenolic anti-inflammatory/anti oxidant compound curcumin, the yellow pigment in turmeric that we found targeted multiple AD pathogenic cascades. The dietary omega-3 fatty acid, docosahexaenoic acid (DHA), also limited amyloid, oxidative damage and synaptic and cognitive deficits in a transgenic mouse model. Both DHA and curcumin have favorable safety profiles, epidemiology and efficacy, and may exert general anti-aging benefits (anti-cancer and cardioprotective.). PMID- 16266773 TI - Insulin resistance syndrome and Alzheimer's disease: age- and obesity-related effects on memory, amyloid, and inflammation. AB - Insulin plays an important role in memory and other aspects of brain function. The insulin resistance syndrome, characterized by chronic peripheral insulin elevations, reduced insulin activity, and reduced brain insulin levels, is associated with age-related memory impairment and Alzheimer's disease (AD). Our work has focused on specific mechanisms through which this association is forged, including the effects of peripheral hyperinsulinemia on memory, inflammation, and regulation of the beta-amyloid peptide that plays a key role in AD pathophysiology. Our data suggest that excessive insulin invokes synchronous increases in levels of Abeta and inflammatory agents, effects that are exacerbated by age and obesity. This constellation of events may have deleterious effects on memory. Treatments focused on preventing or correcting insulin abnormalities may be of therapeutic benefit for adults with age-related memory impairment and AD. PMID- 16266774 TI - Horizontal transmission of the Leningrad-3 live attenuated mumps vaccine virus. AB - Here we describe symptomatic transmission of the Leningrad-3 mumps vaccine virus from healthy vaccinees to previously vaccinated contacts. Throat swab and serum samples were taken from six symptomatic mumps cases and from 13 family contacts. Assessment of serum IgG and IgM anti-mumps virus antibodies and IgG avidity testing was performed using commercial test kits. Sera neutralizing antibodies were measured by plaque reduction neutralization assay using the L-3 vaccine mumps virus as the target. All six of the symptomatic mumps cases and three contact subjects tested positive for mumps by RT-PCR. The genomic sequences tested (F, SH and HN genes) of all nine of these samples were identical to the L 3 mumps vaccine strain. All 13 contacts were asymptomatic; however clear serological evidence of mumps infection was found in some of them. The likely epidemiological source of the transmitted L-3 mumps virus was children who were recently vaccinated at the schools attended by the six symptomatic mumps patients described here. The L-3 mumps vaccine virus can be shed and transmitted horizontally, even to subjects previously vaccinated with the same virus. PMID- 16266775 TI - How many infective viral particles are necessary for successful mass measles immunization by aerosol? AB - OBJECTIVES: This study characterized the performance of the "Classical Mexican Device", which immunized 4 million children against measles, demonstrating the efficacy of aerosol vaccination. METHODS: Using plaque-forming units to quantify virus, the particle size distribution (Next Generation Pharmaceutical Impactor) and rate of output coupled with age specific patterns of breathing allowed the calculation of expected pulmonary deposition. RESULTS: The estimated immunization dose for infants was 30 pfu's, for small children 50, and for older children and adolescents it was 130 and 225, respectively. CONCLUSIONS: These performance characteristics can be used to develop newer battery operable devices that are licensable. PMID- 16266776 TI - Use of a Silastic plate after repair of an oronasal fistula. PMID- 16266777 TI - Enhancing the role of science in the decision-making of the European Union. AB - Used well, science provides effective ways of identifying potential risks, protecting citizens, and using resources wisely. It enables government decisions to be based on evidence and provides a foundation for a rule-based framework that supports global trade. To ensure that the best available science becomes a key input in the decisions made by EU institutions, this abridged version of a working paper produced for the European Policy Centre, a leading, independent think tank, considers how science is currently used in the policy and decision making processes of the EU, what the limitations of scientific evidence are, and how a risk assessment process based on scientific 'good practices' can be advantageous. Finally, the paper makes recommendations on how to improve the use of science by EU institutions. PMID- 16266778 TI - Thirty years of olfactory learning and memory research in Drosophila melanogaster. AB - The last 30 years have witnessed tremendous progress in elucidating the basic mechanisms underlying a simple form of olfactory learning and memory in Drosophila. The application of the mutagenic approach to the study of olfactory learning and memory in Drosophila has yielded insights into the participation of a large number of genes in both the development of critical brain regions as well as in the physiology underlying the acquisition, storage, and retrieval of memory. Newer sophisticated molecular-genetic tools have further allowed for the specification and functional dissection of the neuronal circuitry involved in these processes at a systems level. With these advances in our understanding of the genes, neurons, and circuits involved in learning and memory, the field of Drosophila memory research is nearing a state of integration of the bottom up and top down approaches to understanding this form of behavioral plasticity. PMID- 16266780 TI - A new submerged membrane photocatalysis reactor (SMPR) for fulvic acid removal using a nano-structured photocatalyst. AB - The study focuses on the degradation of fulvic acid by nano-structured TiO2 in a submerged membrane photocatalysis reactor (SMPR). It has been demonstrated that the composite TiO2 photocatalyst could be automatically settled due to its gravity and then be easily separated by MF membrane. In addition, it was more efficient to maintain high flux of membranes than that of commercial TiO2 P25. The paper describes the effects of operational parameters on the photocatalytic degradation of fulvic acid in SMPR. It was found that the photocatalyst at 0.5 g/L and airflow at 0.06 m3/h were the optimal condition for the removal of fulvic acid (FA) and the FA degradation rate was higher at acidic condition than that at alkalinous media. In order to compare the effects of different filtration duration on permeate flux rate of MF, P25 powder and nano-structured TiO2 were employed. According to the experiments, the permeate flux rate of MF is improved and thus the membrane fouling phenomenon is reduced with the addition of nano structured TiO2 catalyst. Therefore, the submerged membrane photocatalysis reactor can be potentially applied in photocatalytic oxidation process during drinking water treatment. PMID- 16266779 TI - Non-genomic effects of endocrine disrupters: inhibition of estrogen sulfotransferase by phenols and chlorinated phenols. AB - Phenols are used world-wide and their presence in the environment is a cause of increasing concern. Despite evidence to suggest that, in general, they bind poorly to estrogen receptors, they are suspected of being endocrine disrupters. Here, we show that 2, x-substituted phenols are potent inhibitors of estrogen sulfotransferase with IC(50) values at low- or sub-micromolar levels. Our results demonstrate a potential non-genomic mechanism of action for these compounds and suggest that, where viable alternatives exist, both phenols substituted in the 2 position and their metabolic precursors should be avoided. PMID- 16266781 TI - Factors affecting adsorption characteristics of Zn2+ on two natural zeolites. AB - Mining-related and industrial wastes are primary sources of heavy metal contamination in soils and groundwater. The limitation of such waste in drinking water needs to meet government requirements in order to safeguard human health and environment. Zinc, one of the most preponderant pollutants, is difficult to remove from wastewater rather than other heavy metals (i.e. lead, copper and cadmium). This paper investigates Zn2+ adsorption characteristics of two natural zeolites found in the regions of Gordes and Bigadic, in western Turkey. The results show that the Zn2+ adsorption behavior of both zeolites is highly dependent on the pH. Adsorption dependence on lower pH values (pH<4) is explained by the dissolution of crystal structure and the competition of the zinc ions with the H+. Between pH 4 and 6, the basic mechanism is the ion exchange process. The results also showed that decrease in grain size does not increase the adsorption capacity of zeolite from Gordes, yet it increases that of zeolite from Bigadic about 23%. The results also reveal that an increase in the initial concentration of Zn2+ in the system causes an increase in the adsorption capacity to a degree, then it becomes more constant at higher concentrations. With this, the removal efficiency of Gordes zeolite is two times higher than that of Bigadic zeolite. Results show that an increase in slurry concentration results in a lower uptake of Zn2+. In the final part of the paper, we compared the experimental data with the Langmuir and Freundlich isotherms. The results show that there is a good fit between the experimental data and empirical isotherms. PMID- 16266784 TI - Measuring the effect of glutamate receptor agonists on extracellular D-serine concentrations in the rat striatum using online microdialysis-capillary electrophoresis. AB - Online microdialysis-capillary electrophoresis (CE) was used to measure the changes in extracellular D-serine concentration in response to the application of glutamate agonists and antagonists in the rat striatum. The microdialysis-CE assay was capable of measuring concentration changes as small as 8% with a sampling rate of 12-15s. Kainic acid (KA) induced increases in D- and L-serine concentrations. Application of the non-NMDA receptor antagonist CNQX did not affect the increases observed for D- or L-serine, suggesting a nonspecific effect. NMDA also induced increases in D-serine, L-serine, glutamate and GABA concentrations. These increases were attenuated by the NMDA receptor antagonist MK-801. PMID- 16266782 TI - Representation of motor skills in human long-term memory. AB - This study uses the example of the tennis serve to investigate the nature and role of long-term memory in skilled athletic performance. Information processing linked with complex movements has always been notoriously difficult to investigate. However, a new experimental method revealed that athletic expertise was characterized by well-integrated networks of so-called basic action concepts (BACs) that each corresponded to functionally meaningful submovements. In high level experts, these representational frameworks were organized in a distinctive hierarchical tree-like structure, were remarkably similar between individuals and were well matched with the functional and biomechanical demands of the task. In comparison, action representations in low-level players and nonplayers were organized less hierarchically, were more variable between persons and were less well matched with functional and biomechanical demands. It is concluded that, in concert with situational goals and constraints, movement representations of this kind in long-term memory might provide the basis for action control in skilled voluntary movements in the form of suitably organized perceptual-cognitive reference structures. PMID- 16266783 TI - Analysis of correlation between serum D-serine levels and functional promoter polymorphisms of GRIN2A and GRIN2B genes. AB - D-Serine is an endogenous coagonist that increases the opening of N-methyl-D aspartate (NMDA)-type glutamate receptor channels. We previously reported a reduction of D-serine serum levels in schizophrenia, supporting the disease hypothesis of NMDA receptor-mediated hypo-neurotransmission. The serum levels of D-serine are thought to reflect brain d-serine content. It is important to understand whether there is a direct link between the altered D-serine levels and NMDA receptor expression in vivo or whether these are independent processes. Two polymorphisms are known to regulate the expression of NMDA receptor subunit genes: (GT)(n) (rs3219790) in the promoter region of the NR2A subunit gene (GRIN2A) and -200T > G (rs1019385) in the NR2B gene (GRIN2B). These polymorphisms are also reported to be associated with schizophrenia. Therefore, we examined the correlation between these two polymorphisms and d-serine serum levels in mentally healthy controls, schizophrenics and the combined group. We observed no significant genotype-phenotype correlations in any of the sample groups. However, analyses of larger sample numbers and the detection of additional polymorphisms that affect gene expression are needed before we can conclude that NMDA receptor expression and serum levels of d-serine, if involved in schizophrenia pathophysiology, are independent and additive events. PMID- 16266785 TI - Colocalization of serotonin and vesicular glutamate transporter 3-like immunoreactivity in the midbrain raphe of Syrian hamsters (Mesocricetus auratus). AB - Vesicular glutamate transporter 3 (VGLUT3) expression has been specifically localized to brain regions rich in serotonergic cells. It has been suggested that this transporter may contribute to the regulation of extracellular glutamate concentrations via a nonsynaptic mechanism. In this study, we examine the colocalization of vesicular glutamate transporter 3 immunoreactivity with serotonin immunoreactivity in the dorsal and median raphe nuclei of Syrian hamsters. Brain sections from adult hamsters were fluorescently labeled for serotonin-ir and VGLUT3-ir and examined using confocal microscopy. The results indicate that most serotonergic cells of the midbrain raphe also expressed vesicular glutamate transporter 3. In addition, nonserotonergic cells in these brain regions also show immunoreactivity for the transporter. These data confirm previous findings of vesicular glutamate transporter 3 expression in serotonergic and nonserotonergic neurons in rats. These findings suggest that the location of vesicular glutamate transporter 3 may be as much a function of neuroanatomical location as of the neurochemical identity of the expressing neurons. PMID- 16266786 TI - Deleting the phosphorylated tail domain of the neurofilament heavy subunit does not alter neurofilament transport rate in vivo. AB - Phosphorylation of the carboxyl tail domains of the neurofilament heavy (NF-H) and middle molecular weight (NF-M) subunits has been proposed to regulate the axonal transport of neurofilaments. To test this hypothesis, we recently constructed mice lacking the extensively phosphorylated NF-H tail domain (NF HtailDelta) and showed that the transport rate of neurofilaments in optic axons is unaltered in the absence of this domain [M.V. Rao, M.L. Garcia, Y. Miyazaki, T. Gotow, A. Yuan, S. Mattina, C.M. Ward, N.A. Calcutt, Y. Uchiyama, R.A. Nixon, D.W. Cleveland, Gene replacement in mice reveals that the heavily phosphorylated tail of neurofilament heavy subunit does not affect axonal caliber or the transit of cargoes in slow axonal transport, J. Cell Biol. 158 (2002) 681-693]. However, Shea et al. proposed that deletion of NF-H carboxyl-terminal region accelerates the transport of a subpopulation of neurofilaments based on minor differences between tail-deleted and control mice in our axonal transport analysis. Here, we present additional evidence that neurofilament transport rate is unchanged after deleting the phosphorylated NF-H tail domain, establishing unequivocally that the NF-H tail domain alone does not regulate the rate of neurofilament transport in optic axons in vivo. Possible roles for tail domains as cross-bridges between a neurofilament and its neighbors or other cytoskeletal elements is discussed. PMID- 16266787 TI - Taxanes in the treatment of early breast cancer. AB - The taxanes docetaxel and paclitaxel have established roles as two of the most active agents in the treatment of metastatic breast cancer. These two drugs are now being incorporated into the management of early breast cancer. A first generation of trials has explored whether the addition of taxanes either sequentially or in combination with adjuvant anthracycline-based chemotherapy improves outcome for patients with early breast cancer. A second generation of trials are now underway which are based on the assumption that taxanes are beneficial in the adjuvant setting, and are comparing the different taxanes, dosing regimens and the addition of further agents. Trials in the neoadjuvant setting have recently demonstrated improved response rates with the addition of taxanes into existing anthracycline-based regimes. This review critically appraises these trials and provides an overview of ongoing research in the area. PMID- 16266788 TI - Childhood behavior problems and peer selection and socialization: risk for adolescent alcohol use. AB - To date, research examining the role of peers in the development of substance use has focused almost exclusively on externalizing behavior problems without considering internalizing behavior problems. This is a notable omission in the literature, because there is some evidence to suggest that internalizing behavior increases risk for substance use, and peers are considered to be among the strongest proximal influences of substance use. The current study considered both internalizing and externalizing behavior problems and examined peer socialization and selection models of alcohol use using a 2-year longitudinal design. We examined potential reciprocal relations between internalizing and externalizing behavior and affiliations with delinquent peers, and how these variables predicted initiation of alcohol use. Participants were 86 children (71% male) ranging from 9-12 years of age (M=10.87). Results were consistent with socialization, whereby delinquent peer affiliations were associated with increases in externalizing behavior, and subsequently early initiation of alcohol use. There was also evidence to suggest that internalizing behavior served as a protective factor for delinquent peer affiliations and for early initiation of alcohol use. Implications of these findings for prevention and intervention efforts are discussed. PMID- 16266791 TI - Management of preterm infants with intrauterine growth restriction. AB - Preterm intrauterine growth restriction (IUGR) is strongly associated with increased mortality and morbidity. In the management of these infants, complications of preterm birth can be amplified by the effect of suboptimal fetal growth. It is important that pregnancies with IUGR are detected before birth, so that delivery can be arranged in a high-risk maternity unit with the appropriate neonatal staff in attendance. The provision of full support for resuscitation and stabilisation of these infants is crucial to the short-term and long-term health of these infants, who have suffered chronic hypoxia and malnutrition in utero. The long term outcome studies of these infants are retrospective and they include SGA infants. The effects of prematurity affect the outcome of IUGR infants. IUGR is associated with cerebral palsy in those delivered more than 32 weeks gestation. Infants less than 32 weeks of gestation may have poor developmental outcome if the head growth is affected, these infants may have associated cognitive and behavioural problems. Children who fail to grow by 2-4 years are at risk of long term growth problems. This paper outlines the acute and long-term management of these infants. PMID- 16266789 TI - More tea for septic patients?--Green tea may reduce endotoxin-induced release of high mobility group box 1 and other pro-inflammatory cytokines. AB - Despite recent advances in antibiotic therapy and intensive care, sepsis remains widespread problems in critically ill patients. The high mortality of sepsis is in part mediated by bacterial endotoxin, which stimulates macrophages/monocytes to sequentially release early (e.g., TNF, IL-1, and IFN-gamma) and late (e.g., HMGB1) pro-inflammatory cytokines. In light of our recent discovery of HMGB1 as a late mediator of lethal systemic inflammation, and the observation that green tea (Camellia sinensis) dose-dependently attenuated bacterial endotoxin-induced HMGB1 release, we propose that regular tea intake might decrease the incidence of and mortality rates from lethal endotoxemia and sepsis. PMID- 16266792 TI - DNA damage induced by ethoxyquin in human peripheral lymphocytes. AB - Ethoxyquin (1,2-dihydro-6-ethoxy-2,2,4-trimethylquinoline, EQ) is widely used in various food products and in animal feeds because of its powerful antioxidant activity. This compound was recently found to cause not only many unfavourable side-effects in animals fed with feeds containing it, but also adverse effects in people exposed to it at work. In the present study, DNA damage induced by EQ in human lymphocytes has been assessed. The alkaline single cell gel electrophoresis assay (comet assay) was used to measure DNA damage. The cells were treated for 1 h with EQ doses ranging from 1 to 250 microM in the absence or in the presence of an exogenous metabolic activation system (S9mix). The obtained results showed that EQ-induced DNA damage in human lymphocytes in a dose-dependent manner; the observed DNA fragmentation induced by EQ in the presence of metabolic activation system was always significantly lower, as compared to cells treated with the same doses of EQ alone. PMID- 16266793 TI - Quality assessment of internet pharmaceutical products using traditional and non traditional analytical techniques. AB - This work investigated the use of non-traditional analytical methods to evaluate the quality of a variety of pharmaceutical products purchased via internet sites from foreign sources and compared the results with those obtained from conventional quality assurance methods. Traditional analytical techniques employing HPLC for potency, content uniformity, chromatographic purity and drug release profiles were used to evaluate the quality of five selected drug products (fluoxetine hydrochloride, levothyroxine sodium, metformin hydrochloride, phenytoin sodium, and warfarin sodium). Non-traditional techniques, such as near infrared spectroscopy (NIR), NIR imaging and thermogravimetric analysis (TGA), were employed to verify the results and investigate their potential as alternative testing methods. Two of 20 samples failed USP monographs for quality attributes. The additional analytical methods found 11 of 20 samples had different formulations when compared to the U.S. product. Seven of the 20 samples arrived in questionable containers, and 19 of 20 had incomplete labeling. Only 1 of the 20 samples had final packaging similar to the U.S. products. The non traditional techniques complemented the traditional techniques used and highlighted additional quality issues for the products tested. For example, these methods detected suspect manufacturing issues (such as blending), which were not evident from traditional testing alone. PMID- 16266794 TI - 3-Farnesyl-2-hydroxybenzoic acid is a new anti-Helicobacter pylori compound from Piper multiplinervium. AB - A new prenylated salicylic acid derivative, 3-farnesyl-2-hydroxy benzoic acid (1), was isolated from the leaves of Piper multiplinervium C. DC. (Piperaceae). It showed anti-Helicobacter pylori activity (MIC 37.5 microg/ml) and antimicrobial activity at MICs between 2.5 and 5 microg/ml against Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Mycobacterium smegmatis, Pseudomonas aeruginosa and Candida albicans. Its structure was elucidated by means of MS, 1H and 13C NMR. The ethnomedical claim of Piper multiplinervium to treat stomach aches by the Kuna Indians of Panama may be justified by anti Helicobacter pylori activity of its MeOH extract. PMID- 16266795 TI - Mechanisms of the vasorelaxant effect of Danshen (Salvia miltiorrhiza) in rat knee joints. AB - The aim of this study is to investigate the mechanisms of the vasorelaxant effect of the crude extract of Salvia miltiorrhiza (family: Labiatae), also known as "Danshen", in rat knee joints. Changes in blood flow of rat knee joints were measured in vivo by a laser Doppler perfusion imager. Topical administration of Danshen onto the exposed knee joint blood vessels produced dose-dependent increases in blood flow. Treatment of the rat knee joint with 2x 1 nmol of atropine, 2x 0.1 nmol of propranolol, or 2x 0.1 nmol of a mixture of pyrilamine plus cimitedine produced no change on the vasodilator response to Danshen. However, significant inhibition of the Danshen-induced vasodilator response was observed in knee joints treated with 2x 100 nmol of N(G)-nitro-L-arginine methyl ester (L-NAME), 2x 100 nmol of flurbiprofen, 2x 10 nmol of the calcitnonin gene related peptide (CGRP) receptor antagonist CGRP(8-37), and also in knee joints but had been denervated by capsaicin treatment or by surgery. Intravenous administration of low doses of Danshen (2.5 and 6 mg/kg) did not affect the systemic blood pressure but significantly increased knee joint blood flow, whereas, high doses of Danshen (167 and 381 mg/kg) produced hypotension with concurrent decreases in knee joint blood flow. These findings indicate that the knee joint blood vessels are more sensitive to the relaxant effect of Danshen compared to blood vessels in the general circulation. The vasorelaxant effect of Danshen was found to be partly mediated by CGRP released from sensory nerves, and nitric oxide and prostaglandins also played a part. However, there is no evidence to support a role for muscarinic receptors, adrenoceptors, or histamine receptors. PMID- 16266796 TI - Collecting duct carcinoma of the kidney: CT and pathologic correlation. AB - PURPOSE: We characterized CT findings of collecting duct carcinoma of the kidney and correlated these with the histopathologic findings. MATERIALS AND METHODS: CT scans of 18 patients with pathologically proven collecting duct carcinoma of the kidney were retrospectively reviewed. We analyzed CT findings of collecting duct carcinoma and also correlated CT findings with the histopathologic findings. RESULTS: The mean size of the tumors was 6.9 cm and all cases were solid. Seventeen (94%) tumors had a medullary location. Nine (69%) and 11 (85%) cases showed weak and heterogeneous enhancement, respectively. A cystic component (50%) was frequently seen within the tumors. Lymphadenopathy and metastasis were noted in 10 (56%) and 6 (33%) cases, respectively. Perinephric stranding and vascular invasion were present in 10 (56%) and 5 (28%) cases, respectively. In 17 (94%) of the 18 cases, involvement of the renal sinus was present. Infiltrative growth (67%) and preservation of the renal contour (61%) were more common than expansile growth (33%) and exophytic configuration (39%), respectively. These CT features were well correlated with the histopathologic findings. CONCLUSION: Medullary location, weak and heterogeneous enhancement, involvement of the renal sinus, infiltrative growth, preserved renal contour, and a cystic component are CT findings frequently seen in patients with collecting duct carcinoma of the kidney. CT findings are nevertheless nonspecific and do not allow collecting duct carcinoma to be easily differentiated from the other subtypes of renal cell carcinoma. However, when CT demonstrates a renal tumor with these findings, collecting duct carcinoma can be considered in the differential diagnosis. PMID- 16266797 TI - Kinetic analysis of the antibacterial activity of probiotic lactobacilli towards Salmonella enterica serovar Typhimurium reveals a role for lactic acid and other inhibitory compounds. AB - Six Lactobacillus strains including commercial probiotic ones (L. acidophilus IBB 801, L. amylovorus DCE 471, L. casei Shirota, L. johnsonii La1, L. plantarum ACA DC 287 and L. rhamnosus GG) were investigated, through batch fermentations under controlled conditions, for their capacity to inhibit Salmonella enterica serovar Typhimurium SL1344. All lactobacilli displayed strong antibacterial activity toward this Gram-negative pathogen and significantly inhibited invasion of the pathogen into cultured human enterocyte-like Caco-2/TC7 cells. By studying the production kinetics of antibacterial activity and applying the appropriate acid and pH control samples during a killing assay, we were able to distinguish between the effect of lactic acid and other inhibitory compounds produced. The antibacterial activity of L. acidophilus IBB 801, L. amylovorus DCE 471, L. casei Shirota and L. rhamnosus GG was solely due to the production of lactic acid. The antibacterial activity of L. johnsonii La1 and L. plantarum ACA-DC 287 was due to the production of lactic acid and (an) unknown inhibitory substance(s). The latter was (were) only active in the presence of lactic acid. In addition, the lactic acid produced was responsible for significant inhibitory activity upon invasion of Salmonella into Caco-2/TC7 cells. PMID- 16266798 TI - Stability and degradation profiles of Spantide II in aqueous solutions. AB - Spantide II is an 11 amino acid peptide that has been shown to be a potential anti-inflammatory agent. The stability and degradation profiles of Spantide II in aqueous solutions were evaluated with the long-term objective of developing topical formulations of this compound for various skin disorders. The stability profile of Spantide II at various temperature and pH conditions was monitored by high performance liquid chromatography (HPLC) and the resulting degradation products were identified by liquid chromatography-mass spectroscopy (LC-MS). Forced degradation of Spantide II was performed at extreme acidic (pH <2.0) and alkaline (pH >10.0) conditions and by addition of hydrogen peroxide (oxidizing agent). The degradation pattern of Spantide II followed pseudo first-order kinetics. The shelf life (T90%) of Spantide II in aqueous ethanol (50%) was determined to be 230 days at 25 degrees C. Spantide II was susceptible to degradation at pH <2 and pH >5 and showed maximum stability at pH 3-5. The stability under various pH conditions indicates that Spantide II was most stable at pH 3.0 with a half-life of 95 days at 60 degrees C. Spantide II degradation was attributed to hydrolysis of peptide bonds [Pro2-(pyridyl)Ala3, (nicotinoyl)Lys1-Pro2, Pro4-PheCl2(5), Trp7-Phe8, Phe8-Trp9, Nle11-NH2), racemization of the peptide fragments that resulted from hydrolysis, cleavage and formation of (nicotinoyl)Lys1-Pro2 diketopiperazine. In the presence of an oxidizing agent, Pro(2,4) residues degraded by ring opening to form glutamyl semialdehyde and by bond cleavage at Pro4 to form 2-pyrrolidone, while Phe(5,8) degraded to form 2-hydroxyphenylalanine. Spantide II was found to be stable in aqueous medium with T90% of 230 days. The major degradation pathways of Spantide II were identified as hydrolysis, racemization, cleavage and formation of diketopiperazine. PMID- 16266799 TI - Both direct and collagen-mediated signals are required for active vitamin D3 elicited differentiation of human osteoblastic cells: roles of osterix, an osteoblast-related transcription factor. AB - In order to investigate the mechanisms by which 1alpha,25(OH)2 vitamin D3 (VD3) stimulates the differentiation of human osteoblasts, we cultured MG-63, which is a human osteoblastic cell line, in the presence or absence of VD3 and/or L ascorbic acid 2-phosphate (Asc 2-P), a long-acting vitamin C derivative. The cell growth rate was decreased by the presence of VD3 in the culture medium. Type I collagen synthesis and alkaline phosphatase (ALP) activity, which are markers of early stage osteoblast differentiation, were stimulated by the presence of VD3 as well as by that of Asc 2-P. The co-presence of Asc 2-P and VD3 had a synergistic effect on the collagen synthesis and ALP activity of the cells. Inhibition of collagen synthesis by the addition of inhibitors of collagen synthesis to the medium attenuated the stimulative effect of VD3 and Asc 2-P on the ALP activity. Transfection of the cells with siRNA-expressing vectors for COL1A1 decreased the expression level of ALP mRNA in addition to that of COL1A1. On the other hand, ALP activity was significantly increased, and the growth rate was decreased, when the cells were cultured on type I collagen-coated dishes. These effects were not seen when the cells were cultured on dishes coated with heat-denatured collagen. VD3 also increased the mRNA levels for Runx2 and osterix, which are transcription factors critical for osteoblast differentiation, as well as those of differentiation markers such as bone/liver/kidney type ALP, COL1A1, (the gene for the alpha1 chain of type I collagen), and osteocalcin, in the cells. Normal human osteoblasts and human bone marrow-derived mesenchymal stem cells (hBMSC) showed quite similar responses to VD3. These results indicate that VD3-stimulated gene expression of type I collagen and that mature type I collagen produced in the presence of Asc 2-P mediates at least a part of the stimulative effects of Asc 2 P and VD3 on the differentiation of these human osteoblastic cells. Levels of mRNAs for ALP and COL1A1 were increased, but the level of Runx2 was decreased, by the expression of osterix in MG-63 cells. These results also suggest that VD3 controls the growth and differentiation of human osteoblastic cells by regulating the gene expression of osteoblast-related transcription factors as well as that of type I collagen, and that the co-presence of both signals is essential for VD3 to express full activity toward the differentiation of human osteoblasts. PMID- 16266800 TI - Needle-track metastases and prophylactic radiotherapy for mesothelioma. AB - BACKGROUND: Mesothelioma invades the tracts made by chest instrumentation. Prophylactic radiotherapy is effective at preventing malignant seeding at these sites. METHODS: We assessed the use and effectiveness of radiotherapy at our centre in 39 of the 40 patients identified with mesothelioma between January 2000 and September 2003. RESULTS: Thirty-seven (95%) patients received radiotherapy to their chest instrumentation site between 6 and 42 days (median 26 days) following the diagnosis of mesothelioma. The radiotherapy field size varied, from 4 cm square to 14 x 10.5 cm. The radiotherapy was given as 21 Gy in 3 fractions over 1 week. In 3 patients (8%), there was already tumour invasion of the skin at the time of radiotherapy. In 2 other patients (5%), there was tumour recurrence following radiotherapy; in both this was at the edge of the previous radiotherapy fields. Further treatment was administered to an adjacent field in both. One patient with an indwelling pleural catheter developed tumour growth at the catheter insertion site. This was treated successfully with radiotherapy, with no catheter damage. CONCLUSIONS: Prompt radiotherapy referral and radiotherapy field selection is important to maximise the effect of radiotherapy given to prevent chest wall tumour growth. There was no tumour growth in areas that were treated with radiotherapy. Further chest interventions outside the radiotherapy field should be followed with further radiotherapy. PMID- 16266801 TI - Cough frequency, cough sensitivity and health status in patients with chronic cough. AB - BACKGROUND: Little is known about the frequency of cough in health and in patients with chronic cough. METHODS: We measured cough frequency and its relationship with other markers of cough severity in 20 patients with chronic cough and 9 healthy subjects using the Leicester Cough Monitor (LCM), which is an automated ambulatory digital cough monitor that records sound only. All subjects had a 6-h recording and recordings were manually counted. A subgroup of 6 normals and 6 patients with a stable chronic cough had repeat measurements up to 6 months apart. RESULTS: Mean (sem) cough counts/hour were 43(8) in patients with chronic cough and 2(1) in normals (mean difference 41; 95% confidence interval 24-59; P<0.001). The cough counts were repeatable (within subject standard deviation: 23 coughs/hour; intraclass correlation coefficient 0.8). Cough counts correlated significantly with physical (r=-0.6, P=0.03), social (r=-0.7, P=0.01) and total Leicester Cough Questionnaire (LCQ) health status scores (r=-0.6, P=0.03) and cough sensitivity (concentration of capsaicin causing 5 coughs: r=0.9, P=0.008). CONCLUSION: We have shown that there are marked differences in cough frequency between patients with chronic cough and healthy subjects, that these measurements are repeatable, and that they correlate with cough-specific health status. PMID- 16266802 TI - An evolutionary perspective on FoxP2: strictly for the birds? AB - FoxP2 mutations in humans are associated with a disorder that affects both the comprehension of language and its production, speech. This discovery provided the first opportunity to analyze the genetics of language with molecular and neurobiological tools. The amino acid sequence and the neural expression pattern of FoxP2 are extremely conserved, from reptile to man. This suggests an important role for FoxP2 in vertebrate brains, regardless of whether they support imitative vocal learning or not. Its expression pattern pinpoints neural circuits that might have been crucial for the evolution of speech and language, including the basal ganglia and the cerebellum. Recent studies in songbirds show that during times of song plasticity FoxP2 is upregulated in a striatal region essential for song learning. This suggests that FoxP2 plays important roles both in the development of neural circuits and in the postnatal behaviors they mediate. PMID- 16266803 TI - Side-chain modified analogues of histaprodifen: asymmetric synthesis and histamine H1-receptor activity. AB - New analogues of histaprodifen with polar side chains have been stereoselectively synthesized and evaluated as histamine H(1)-receptor agonists. As a key transformation the asymmetric aminohydroxylation has been used, which was successfully realized for the first time on an imidazolyl derivative. While all chiral analogues proved to be weak H(1)-receptor antagonists, an achiral keto derivative of histaprodifen turned out to be the first 2-acylated histamine congener displaying partial H(1)-receptor agonism (relative potency 12%). PMID- 16266804 TI - Molecular modeling analysis of the interaction of novel bis-cationic ligands with the lipid A moiety of lipopolysaccharide. AB - Lipopolysaccharides (LPS), otherwise termed "endotoxins", are outer-membrane constituents of Gram-negative bacteria and play a key role in the pathogenesis of "septic shock", a major cause of mortality in the critically ill patient. We have shown that the pharmacophore necessary for optimal recognition and neutralization of LPS by small molecules requires an interaction between two protonatable positive charges separated by a distance of approximately 14A, which corresponds to the distance between two anionic phosphates on the glycolipid component of LPS called lipid A. The in silico binding of a diverse set of compounds with bis amino, -amidino, -guanidino, and -aminoguanidino functionalities, identified as potential lead scaffolds in a high-throughput screen, with lipid A was explored using molecular docking simulations. A weighted expression for binding affinity was trained relative to experimental ED(50) measurements, attaining a correlation of R(2)=0.66. Our docking results showed that the electrostatic interaction between ligands and lipid A phosphates dominates the expression and varies little across the series, and other ligand-receptor interactions seem to play a secondary role in governing the observed variations in the relative ligand binding affinity. Further, it appears that the ligand internal energy plays the primary role in differentiating between compound binding affinities which also correlated well with experimental ED(50) data (R=0.77). Application of this strategy would be useful in the de novo design of highly active endotoxin sequestering agents. PMID- 16266805 TI - Inactivation of bovine plasma amine oxidase by haloallylamines. AB - Various 2- and 3-haloallylamines were synthesized and evaluated as inhibitors of the quinone-dependent bovine plasma amine oxidase (BPAO). 3-Haloallylamines, which were previously found to be good inhibitors of the flavin-dependent mitochondrial monoamine oxidase (MAO), exhibited a time-dependent inactivation of BPAO, with the 2-phenyl analogs being more potent than the 2-methyl analogs. No plateau of enzyme activity loss was observed, suggestive of a lack of competitive partitioning to normal turnover. The (E)- and (Z)-2-phenyl-3-fluoro analogs were the most potent (low microM IC(50)s), with the corresponding 3-bromo and 3-chloro analogs being >10-fold less potent. In each case, the Z-isomers were more potent than the E-isomers, the reverse of the configurational inhibitory preference observed with MAO. In contrast to the 2-phenyl analogs, 3-phenyl-2(or 3) chloroallylamines displayed a partitioning behavior, consistent with these being both substrates and inactivators of BPAO. PMID- 16266806 TI - DFT study of the reductive decomposition of artemisinin. AB - Artemisinin is a sesquiterpene lactone with an endoperoxide function that is essential for its antimalarial activity. The DFT B3LYP method, together with the 6-31G(d) and 6-31+G(d,p) basis set, is employed to calculate a set of radical anions and neutral species supposed to be formed during the rearrangement of artemisinin from the two radicals (C-centered and O-centered) that are supposed to play a relevant role in the mechanism of action. The B3LYP results show that the primary and the secondary radicals centered on C(4), generated by homolytic break of the C(3)-C(4) bond and by 1,5 hydrogen shift, respectively, are more stable than radicals centered on oxygen. The calculations show that the activation barriers for rearrangements are low, leading to a thermodynamically favorable process. These results reinforce our previous conclusions based on semi empirical calculations but also give additional information on the reductive decomposition of artemisinin. PMID- 16266807 TI - Synthesis of hypermodified adenosine derivatives as selective adenosine A3 receptor ligands. AB - We investigated the A(3)AR affinity and selectivity of a series of 2-substituted 3'-azido and 3'-amino adenosine derivatives as well as some 5'-uronamide derivatives thereof. All compounds showed high A(3)AR selectivity. While the 3' azides appeared to be A(3)AR antagonists with moderate A(3)AR affinity, their 3' amino congeners exhibit significantly improved A(3)AR affinity and behave as partial agonists. For both the 3'-azides and the 3'-amines, the 5' methylcarbamoyl modification improved the overall affinity. Introduction of a 2 phenylethynyl substituent provided high affinity for the A(3)AR. PMID- 16266808 TI - Structure-intrinsic activity relationship studies in the group of 1-imido/amido substituted 4-(4-arylpiperazin-1-yl)cyclohexane derivatives; new, potent 5-HT1A receptor agents with anxiolytic- like activity. AB - Introduction of 1,4-disubstituted cyclohexane ring in the structure of flexible long chain arylpiperazines resulted in linearly constrained, potent serotonin (5 HT)(1A) ligands. In order to trace structure-intrinsic activity relationships in this group, a new series of 1-substituted 4-(4-arylpiperazin-1-yl)cyclohexane derivatives with different cyclic imide/amide termini, and their flexible, tetramethylene analogues were synthesized and pharmacologically evaluated for 5 HT(1A) receptors. In vitro binding experiments revealed that all the compounds were potent 5-HT(1A) receptor agents (K(i) = 1.9-74 nM). Some derivatives tested additionally showed also high affinity for alpha(1)-adrenergic receptors (K(i) = 2.9-101 nM) and for 5-HT(7) receptors. Functional in vivo examination revealed that rigid ligands with o-OCH(3) group in the aryl moiety and cyclic imide system in the opposite terminal behaved like postsynaptic 5-HT(1A) receptor antagonists. On the other hand, unsubstituted, m-Cl, or m-CF(3) substituted derivatives as well as those with cyclic amide group in the terminal fragment exhibited agonistic or partial agonistic activity. Three out of four derivatives tested, that is, postsynaptic 5-HT(1A) antagonists 9 and 10, and partial agonist 16, showed anxiolytic-like activity in the conflict drinking (Vogel) test in rats. PMID- 16266810 TI - The acute alterations in biochemistry, morphology, and contractility of rat isolated terminal ileum via increased intra-abdominal pressure. AB - AIM AND SCOPE: To determine the acute effects of increased intra-abdominal pressure (IAP) on the biochemistry, morphology, and contractility of the isolated terminal ileum of rats. BACKGROUND: Laparoscopic procedures are used clinically in diagnostic and treatment modalities and experimentally as a model of ischemia reperfusion injury induced by the elevation of IAP. Although some clinical and in vivo experimental studies investigate the results of ischemia-reperfusion injury whether induced by elevated IAP or clamping, there is no in vitro study that has investigated the acute effects of high IAP mimicked by a laparoscopic intervention in any of the intra-abdominal organs (like terminal ileum) on the basis of contractility which represents the motility. METHODS: Twenty-four adult with either sex Sprague-Dawley rats were divided into three groups. The control group (Group I) was not subjected to any IAP. In Groups II and III, an IAP of 10 and 20 mmHg, respectively, was established by carbon dioxide pneumoperitoneum for a period of 60 min. Thirty minutes after the desufflation, the terminal ileum was removed for in vitro pharmacological investigation, measurement of malondialdehyde (MDA) values, and histopathological examination. Statistical comparisons among groups were done using the Kruskal-Wallis variance analysis, with post hoc comparison performed with the Mann-Whitney U-test. RESULTS: Tissue MDA value and the damage scores of mucosa and submucosa were significantly increased in both IAP groups. The smooth muscle layer was significantly damaged only in Group III. The contractions obtained by electrical field stimulation (EFS) were inhibited in both IAP groups, and the contractions to acetylcholine were inhibited in Group III when compared to the control group. CONCLUSIONS: In conclusion, we can say that pneumoperitoneum induced IAP may inhibit contractile responses, cause structural alterations which may be related to ischemia reperfusion injury in rat terminal ileum. PMID- 16266809 TI - Identification of 12Cysbeta on tubulin as the binding site of tubulyzine. AB - We have undertaken quantitative binding site studies in order to identify the binding site of the known microtubule destabilizing agents, the tubulyzines, in the tubulin dimer. Two different approaches were employed that utilized the tubulyzines and their derivatives. The first approach was based on a chemical affinity labeling method using tubulyzine affinity derivatives, and the second approach employed the mass spectrometric measurement of the differential reactivity of cysteines using the tubulyzines and monobromobimane. Based on overlapping data from these two approaches, we propose that the tubulyzines bind at the guanosine-5'-triphosphate binding site of beta-tubulin. Interestingly, we also show that the tubulyzines' binding to tubulin induces a conformational change in tubulin that prevents further interaction of the 239Cysbeta with other reagents. PMID- 16266812 TI - Does level of load affect relative activation levels of vastus medialis oblique and vastus laterialis? AB - The purpose of the study was to evaluate the effect of different relative loading levels on the EMG activity of Vastus Medialis Oblique (VMO) and Vastus Laterialis (VL). Previous research into the EMG temporal and spatial relationship between VMO and VL has increased the controversy surrounding the topic, due to the majority of studies failing to be consistent in electrode placement, level of loading and subject selection. It is generally believed that the nature of the loading task will significantly affect results; despite this few studies have controlled relative load level between subjects. EMG activity of VMO and VL was measured at four load levels (MIVC, 75%, 50% and 25% of MIVC) in 10 asymptomatic male subjects. No difference in onset of activity was found between VMO and VL (p>0.05) and onset of activity was not affected by level of load (p>0.05). The relative level of load had a significant effect both on overall activity of VMO and VL, and the ratio of their activity. The study has shown that relative level of load can have significant effects on the parameters measured and if this variable is not controlled for within the study design it becomes a potential confounding effect. PMID- 16266811 TI - A dynamic view of the immunological synapse. AB - T cell activation requires interactions of T cell antigen receptors (TCR) and peptides presented by major histocompatibility complex molecules (MHCp) in an adhesive junction between the T cell and antigen-presenting cell. Stable junctions with bull's eye supramolecular activation clusters (SMACs) have been defined as immunological synapses (IS). These structures maintain T cell-APC interaction and allow directed secretion. T cells can also be activated by asymmetric hemi-synapses (HS) that allow migration during signal integration. IS and HS operate in different stages of T cell priming. Optimal effector functions may also depend upon cyclical use of IS and HS. PMID- 16266813 TI - Molecular phylogenetics and biogeography of the nacellid limpets of New Zealand (Mollusca: Patellogastropoda). PMID- 16266814 TI - Molecular phylogenetic relationship of toads distributed in the Far East and Europe inferred from the nucleotide sequences of mitochondrial DNA genes. AB - The toads of the Bufo bufo species group are widely distributed in the Eurasian continent and Japanese Archipelago. In this study, we analyzed the mtDNA gene sequences of this species group and estimated the divergence time to clarify the evolutionary relationships and biogeography of toads distributed in the Far East and Europe. The phylogenetic tree indicated that this group produced Bufo bufo in Europe, whereas it produced B. japonicus in the Far East. B. japonicus was divided into three major clades corresponding to a group consisting of B. j. gargarizans in China, B. j. bankorensis in Taiwan, and B. j. miyakonis on Miyako Isl. and eastern and western groups of Japanese B. j. japonicus subspecies group. The eastern and western groups were divided into several subclades which tended to reflect the region-specific geographic distribution of all localities except B. j. japonicus from Hakodate. The estimated branching times of these clades suggest that geological events may have influenced the divergence of the toads distributed in the Far East and Europe. PMID- 16266815 TI - Characterisation of the mating-type locus in the genus Xanthoria (lichen-forming ascomycetes, Lecanoromycetes). AB - Conserved regions of mating-type genes were amplified in four representatives of the genus Xanthoria (X. parietina, X. polycarpa, X. flammea, and X. elegans) using PCR-based methods. The complete MAT locus, containing one ORF (MAT1-2-1) coding for a truncated HMG-box protein, and two partial flanking genes, were cloned by screening a genomic lambda phage library of the homothallic X. parietina. The flanking genes, a homologue of SLA2 of Saccharomyces cerevisiae and a DNA lyase gene, served to amplify the two idiomorphs of the X. polycarpa MAT locus. Each idiomorph contains a single gene: MAT1-2-1 codes for a HMG-box protein, MAT1-1-1 encodes an alpha domain protein. The occurrence of mating-type genes in eight single spore isolates derived from one ascus was studied with a PCR assay. In the homothallic X. parietina a HMG fragment, but no alpha box fragment was found in all isolates, whereas in X. elegans, another homothallic species, all tested isolates contained a fragment of both idiomorphs. Conversely, isolates of the heterothallic X. polycarpa contained either a HMG or an alpha box fragment, but never both. PMID- 16266816 TI - Application of a KDPG-aldolase gene-dependent addiction system for enhanced production of cyanophycin in Ralstonia eutropha strain H16. AB - Two different recombinant plasmids both containing the cyanophycin synthetase gene (cphA) of Synechocystis sp. strain PCC6308 but differing concerning the resistance marker gene were tested for their suitability to produce high amounts of cyanophycin in recombinant strains of Ralstonia eutropha. Various cultivation experiments at the 30-L scale revealed very low cyanophycin contents of the cells ranging from 4.6% to 6.2% (w/w) of cellular dry weight (CDW) only, most probably because most cells had lost the corresponding plasmid during cultivation. To establish a cost effective and high efficient system for production of cyanophycin at larger scales using recombinant strains of R. eutropha, we applied two strategies: First, we integrated cphA into the dispensable chromosomal l lactate dehydrogenase gene (ldh) of R. eutropha. Depending on the cultivation conditions used, relatively low cyanophycin contents between 2.2% and 7.7% (w/w) of CDW were reproducibly detected, which might be due to weak expression or low gene dosage in the single cphA copy strain of R. eutropha. In a second strategy we constructed a KDPG-aldolase gene (eda)-dependent addiction system, which combined features of a multi-copy plasmid with stabilized expression of cphA. Flasks experiments revealed that the cells accumulated extraordinarily high amounts of cyanophycin between 26.9% and 40.0% (w/w) of CDW even under cultivation conditions lacking cyanophycin precursor substrates or plasmid stabilizing antibiotics. Cyanophycin contents of up to 40.0% (w/w) of CDW were also obtained at a 30-L scale or a 500-L pilot-plant scale under such non selective conditions. This demonstrates impressively that the stabilizing effect of the constructed eda-dependent addiction system can be used for production of enhanced amounts of cyanophycin at a larger scale in recombinant strains of R. eutropha. PMID- 16266817 TI - Head and neck cancer cell lines exhibit differential mitochondrial repair deficiency in response to 4NQO. AB - Constituents of tobacco can cause DNA adduct formation and are implicated in head and neck squamous cell cancer (HNSC) development. We investigated the capacity of HNSC cell lines to repair mitochondrial DNA (mtDNA) damage induced by a DNA adduct-forming agent. HNSC cell lines underwent 4-nitroquinoline 1-oxide (4NQO) exposure with subsequent rescue with normal media. Real-time quantitative PCR for nuclear DNA (nDNA) and mtDNA was performed. mtDNA to nDNA ratios were calculated and standardized to mock-treated cells to assess mtDNA repair ability. Two of three tested cancer cell lines exposed to 4NQO exhibited consistent decreases in mtDNA/nDNA ratios throughout the different repair timepoints. At 24 h mtDNA/nDNA ratios of JHU-O19 and JHU-O22 decreased to 63% and 60% of controls, respectively. Conversely, a control keratinocyte cell line exhibited overall increases in mtDNA/nDNA ratios compared to baseline suggesting intact DNA repair mechanisms. By using a DNA adduct formation and repair model featuring 4NQO and HNSC cell lines, we have implicated faulty mtDNA repair as having a potential role in HNSC. PMID- 16266818 TI - Photodynamic therapy with mono-L-aspartyl chlorin e6 can cause necrosis of squamous cell carcinoma of tongue: experimental study on an animal model of nude mouse. AB - Mono-L-aspartyl chlorin e6 (NPe6) is an effective photosensitizer with a major absorption band at 664 nm. NPe6 is potentially exploitable for photodynamic therapy (PDT) and does not cause the side effect of prolonged normal skin photosensitization. However, there are no clinical and experimental reports of its use in oral cancer till now. In the present study, we examined the effectiveness of NPe6-induced PDT with a diode laser for treatment of tongue cancer in the nude mouse. Six nude mice with experimental tongue cancer (HSC-3) were given 10 mg/kg NPe6 intravenously. Two hours later PDT was performed using a laser diode at a light dose of 100 J/cm2 and wavelength of 664 nm. Histological changes in the tumors were examined 42-72 h after PDT. Almost all of the tumors developed necrosis, while viable-like neoplastic cells remained mainly in the peripheral region of the tumor in some cases. The mean depth of necrosis below the surface was 2.1 mm. The mean tumor thickness below the surface was 2.3 mm. Tumor thickness coincided with the depth of necrosis. NPe6-induced PDT exhibited tumor selectivity and can effectively cause necrosis of tongue cancers. This therapy could be suggested for treatment of other superficial oral cancer. PMID- 16266819 TI - Growth factor-sensitive molecular targets identified in primary and metastatic head and neck squamous cell carcinoma using microarray analysis. AB - Polypeptide growth factors play key roles in the processes of cell migration and invasion. In this study, we have used cDNA microarrays to identify target genes whose expression is differentially modulated by the growth factors TGFbeta and EGF. HN4 and HN12 cell lines, established from primary tumor and a lymph node metastasis arising in one patient with head and neck squamous cell carcinoma, were treated with 2nM EGF or 50pM TGFbeta for 24h and extracted RNA was used to prepare labeled cDNAs which were hybridized to NCI UniGem 2.0 cDNA microarrays containing 9128 features. Results revealed constitutive overexpression of 41 genes and underexpression of 109 genes in metastatic HN12 compared to HN4 under conditions of serum withdrawal. Furthermore, TGFbeta treatment resulted in relative upregulation of 53 genes and downregulation of 91 genes in HN12 compared with HN4, whereas cells treated with EGF showed relative upregulation of 67 genes and downregulation of 113 genes. Partial overlap was found between TGFbeta and EGF-modulated gene sets. Results were verified for a subset of each category using quantitative PCR, western blotting and zymography. The data indicate that TGFbeta and EGF differentially affect gene expression in primary and metastatic HNSCC cells, and likely contribute to the invasive properties of metastatic cells through regulation of both common and specific mediators for each growth factor. PMID- 16266820 TI - Screening for distant metastases in patients with head and neck cancer: is there a role for (18)FDG-PET? AB - The detection of distant metastases and second primary tumours at the time of initial evaluation changes the prognosis and influences the selection of treatment modality in patients with HNSCC. Until recently chest CT was the single most effective test to screen for distant metastases in HNSCC patients. In this observational cohort study we prospectively compared the yield of whole body (18)FDG-PET and chest CT to detect distant metastases and synchronous primary tumours. The results of whole body (18)FDG-PET and chest CT were analysed in 34 consecutive HNSCC patients with previously established risk factors for the presence of distant metastases. Four patients were diagnosed with distant metastases or second primary tumours: CT as well as (18)FDG-PET identified one patient with lung metastases and another with primary lung cancer. In addition, (18)FDG-PET detected second primary tumours in two patients (hepatocellular carcinoma and abdominal adenocarcinoma). However increased uptake sites at (18)FDG-PET in lung, liver and pelvis in five patients were not confirmed by other imaging modalities. The added value of whole body (18)FDG-PET versus chest CT was to identify unknown malignancy in 6% of the patients. Confirmation of positive (18)FDG-PET findings is feasible and necessary. PMID- 16266821 TI - Epithelioid hemangioma in the oral mucosa: a clinicopathological study of seven cases and review of the literature. AB - To investigate the clinicopathological characteristics and biologic behavior of epithelioid hemangioma in oral mucosa, the clinical features and pathological findings of seven cases of epithelioid hemangioma were reviewed, and all sections were examined by immunohistochemistry. There were 4 males and 3 females ranged from 8 to 65 years (mean 39.7 years). Sites of origin included the tongue (n = 4), lips (n = 2), and palates (n = 1). Five lesions presented as a solitary mass. Three patients had history of minimal trauma. Histopathologically, well-formed but often immature vessels lined by plump epitholid endothelial cells with prominent inflammatory components were observed. Immunohistochemical staining revealed that tumour cells were positive for Vimentin, FVIIIRAg and CD34. All the cases were negative to S-100 protein and EMA. One case recurred after surgical excision, while none of them metastasized. Epithelioid hemangioma is a benign vascular anomaly, and has close relationship with local trauma. The differential diagnosis is usually based upon histopathological findings. PMID- 16266822 TI - Animal models in oral cancer research. AB - Biologically and clinically relevant animal models are essential in investigation of the progression of diseases and the elaboration of diagnostic or therapeutic protocols. The several rodent models used for in vivo evaluation for oral cancer employ chemical, transplantation and genetic (knockout and transgenic) induction methods. These models are described together with their advantages and disadvantages. Their optimization and application in future research may improve the early detection and treatment of oral cancer. PMID- 16266824 TI - Factors regarding increase of platelet counts in chronic hepatitis C patients with sustained virological response to interferon-Relation to serum thrombopoietin levels. AB - Thrombocytopenia is frequently found in patients with chronic liver disease, and associated with advanced fibrosis stage and with decreased liver function. Serum thrombopoietin (TPO) levels also decrease as the disease progresses from mild fibrosis to cirrhosis. On the other hand, platelet counts increase associated with improvement of fibrosis in chronic hepatitis C (CH-C) patients with sustained virological response (SVR) to interferon (IFN) therapy. Then, we studied if the increase of platelet counts in SVR associate with elevated TPO production or a reduction of spleen size. Liver fibrosis, spleen size, serum TPO levels, albumin, zinc turbidity test (ZTT), platelet counts were compared in fifteen CH-C patients with SVR before and after IFN therapy. RESULTS: Albumin increased from 4.2+/-0.3 to 4.3+/-0.3g/dl (p=0.067), ZTT decreased from 17.7+/ 5.9 to 8.9+/-3.9K-U (p<0.001), platelet counts increased from 15.5+/-6.8x10(4) to 19.9+/-5.8x10(4)/mul (p<0.01) and serum TPO levels increased from 1.65+/-0.94 to 2.06+/-1.22fmol/ml (p=0.073). Spleen size was measured by ultrasonography, and the spleen index was calculated by multiplication of the long and short axes from hilus, which decreased from 14.6+/-5.0 to 10+/-3.1 (p<0.001) after IFN therapy. In conclusion, increase of platelet counts in SVR may be related to the reduction of spleen size and increased serum TPO levels associated with improvement of fibrosis after IFN therapy. PMID- 16266823 TI - Prevalence of hepatitis C virus antibodies among blood donors in north-eastern Poland. AB - In many countries, screening blood donors for viral markers is an important source of information about epidemiology of HCV infection. The aim of this study was to determine seroprevalence of HCV infection among blood donors in north eastern Poland and to compare it with prevalence of markers of other infections. We retrospectively analysed the results of tests for anti-HCV, HBsAg and anti-HIV of all blood donations performed in years 1998-2003. For HCV infection, full data (including all results of confirmatory tests) were available only for years 1998 2000. Overall prevalence of anti-HCV among all donors in years 1998-2000 was 0.5%. There was no significant difference in anti-HCV positivity rate of male and female donors. Majority of HCV infections occurred among first time donors (152/202; 75% of all HCV-positive results). The prevalence of anti-HCV among first time donors averaged 0.6% and remained at similar level as HBsAg (0.7%). The number of anti-HIV positives among first time donors remained low (mean prevalence 0.01%). We conclude that prevalence of anti-HCV among blood donors in podlaskie woiewodship is similar to prevalence in Poland but higher than reported for Western European countries and USA. PMID- 16266825 TI - Clinical determinants of poor six-minute walk test performance in patients with left ventricular systolic dysfunction and no major structural heart disease. AB - BACKGROUND: The clinical determinants of six-minute walk test (6-MWT) performance in patients with left ventricular systolic dysfunction (LVSD) have rarely been investigated, and it is not clear whether they differ from patients referred for the assessment of symptoms of heart failure who do not have major structural heart disease (MSHD). METHODS AND RESULTS: 571 patients with LVSD enrolled in a chronic disease management programme (79% male; mean age 71+/-10 years; BMI 28+/ 5) completed a 6-MWT with a mean distance 337+/-103 m. 688 patients referred with suspected heart failure but in whom MSHD was excluded (49% male; mean age 70+/-11 years; BMI 28+/-6) had a mean 6-MWT distance of 391+/-106 m (P<0.001 compared to patients with LVSD). Relationships with walking distance were determined by calculating odds ratios (ORs) with 95% confidence intervals (CIs) for walking300 m. In patients with LVSD, predictors of poor walking distance (or=75 years (OR=4.0, 95% CI=2.4-6.4); low BMI (<20) (OR=3.4, 95% CI=1.6-7.3); anaemia (OR=2.8, 95% CI=1.8-4.2); resting heart rate>80 beats x min(-1) (OR=2.2, 95% CI=1.3-3.5); and being female (OR=2.0, 95% CI=1.3-3.0). Serum creatinine and NT-proBNP showed dose-response effects, as did self-perceived feelings of depression and anxiety. Determinants of 6-MWT in patients without MSHD were similar including age>or=75 years (OR=6.0, 95% CI=3.4 10.4), anaemia (OR=2.8, 95% CI=1.6-4.9), resting HR>80 beats x min(-1) (OR=2.5, 95% CI=1.4-4.4) and being female (OR=1.6, 95% CI=1.9-2.4). NT-proBNP and self perceived feelings of depression and anxiety also showed dose-response effects. CONCLUSION: The determinants of poor 6-MWT performance depend on physical cardiovascular and non-cardiovascular, and psychological factors. Clinical predictors for poor walking performance are similar for patients with LVSD and without MSHD. PMID- 16266826 TI - EpiTrack: tracking cognitive side effects of medication on attention and executive functions in patients with epilepsy. AB - RATIONALE: Achievement of maximum seizure control with preservation or even improvement of patient's cognitive capabilities is the major aim of epilepsy therapy. EpiTrack is a brief screening tool for the tracking of cognitive side effects of antiepileptic drugs. Test selection was based on recent studies on the effects of topiramate on cognition and retrospective inspection of results from patients with antiepileptic drug (AED) side effects. METHODS: The 15-minute screening tool comprises six subtests: the Trail-Making Test (parts A and B), a test of response inhibition, digit span backward, written word fluency, and a maze test. These tests were standardized in 220 healthy subjects, 100 of whom were reevaluated after 5.3 months to obtain information on reliability and practice effects. Criterion validity was determined by correlation to other neuropsychological measures. For a first clinical evaluation, the impact of epilepsy (seizures) and medication on EpiTrack scores was evaluated cross sectionally in 184 consecutive inpatients with chronic epilepsy. RESULTS: According to the normative data, we developed an easy scoring scheme assigning test scores on a 7-point scale. The EpiTrack is suitable for patients between 18 and 60 years of age. Age corrections were included for patients between 40 and 60 years. EpiTrack scores on subtests for both controls and patients were submitted to principal component analysis. VARIMAX rotation yielded a two-factor solution (verbal/visuo-spatial) that accounted for 63.8% of the total variance in controls. In the patient group, only one factor emerged accounting for 54.7% of variance. EpiTrack correlates with global scores of attention (r=0.85) and language (r=0.67) (P's<0.001). At a cutoff score of 25, only 2.7% of the controls were classified as impaired, while impairment was indicated in 48.4% of the patients. The score is sensitive to monthly frequency of complex partial seizures and to number of AEDs. It shows negative cognitive effects of valproate and topiramate given in mono/polytherapy. CONCLUSION: EpiTrack is a promising 15 minute screening tool for the detection and tracking of cognitive side effects of AEDs and adverse effects of seizures in patients with epilepsy. Future application will show its value in prospective follow-up studies on AED side effects. PMID- 16266827 TI - Neurocognitive effects of phenobarbital discontinuation in epileptic children. AB - PURPOSE: Phenobarbital (PB) is the most widely used antiepileptic drug in the world, but its possible deleterious cognitive and behavioral side effects remain an important concern among physicians and patients.We therefore investigated whether discontinuation of PB in children with epilepsy is accompanied by improvement in cognitive function. METHODS: Neuropsychiatric performance was evaluated with the Wechsler Intelligence Scale for Children-Revised (WISC-R), in two consecutive 7-month periods, in two groups of children with epilepsy 6-12 years old who had been seizure-free for at least 2 years and whose only antiepileptic drug was PB. The case group comprised 24 patients who discontinued PB, and the control group was comprised of the 21 children who continued to take PB. RESULTS: Discontinuation of PB improved Total IQ in the case group compared to the control group (P = 0.027). This increase was mostly in performance (nonverbal) items; verbal items remained almost unchanged. CONCLUSION: These findings suggest that PB affects cognitive function, and the performance (nonverbal) deficits are diminished after discontinuation of therapy. PMID- 16266828 TI - Reelin mouse mutants as models of cortical development disorders. AB - Developmental defects in neuronal positioning and synaptic connectivity are commonly found in neurological diseases, and they are believed to underlie many cognitive and affective disorders. Several mouse mutants are currently available that model at least some aspects of human developmental brain disorders. With the identification of the genes mutated in these animals and the study of the cellular basis of the phenotypes, we have taken significant strides toward an understanding of the mechanisms controlling proper brain development and the consequences of their dysfunction. In particular, mouse mutants deficient in the Reelin gene have provided valuable insights into the mechanisms of cortical development. Absence of Reelin expression in the spontaneous mutant mouse reeler leads to extensive defects in neuronal position and dendrite development. In humans, loss of Reelin results in a type of lissencephaly with severe cortical and cerebellar malformation. Genetic and biochemical studies using mouse mutants suggest that the Lis1 protein may participate in the Reelin signaling pathway controlling cortical development. Reduced levels of Reelin are also present in postmortem brains of patients with schizophrenia, suggesting a possible link with this cognitive disorder. The regulation of the Reelin gene may thus provide insights into the mechanisms of this disease. PMID- 16266829 TI - Older adults with epilepsy demonstrate cognitive impairments compared with patients with amnestic mild cognitive impairment. AB - Little is known about the cognitive effects of chronic epilepsy in older adults. To better characterize cognitive impairment in seniors with epilepsy, we compared cognitive performance of 26 seniors with epilepsy with that of 26 well-matched patients with mild cognitive impairment (MCI) and 26 well-matched healthy older adults. Participants completed neuropsychological testing with the Dementia Rating Scale (DRS), Logical Memory, and CFL Word Fluency. There were no significant demographic group differences, although seniors with epilepsy had higher self-reported depression. Seniors with epilepsy performed below controls on virtually all neuropsychological tests, and performed below patients with MCI on DRS Total score, Initiation/Perseveration, and CFL Fluency. Seniors with epilepsy on antiepileptic drug (AED) polytherapy had the most severe cognitive deficits, whereas seniors with epilepsy on AED monotherapy were comparable to cholinesterase inhibitor-naive patients with MCI. This study emphasizes the clinical importance of cognitive impairment in seniors with epilepsy and highlights the need for future studies addressing causes and treatment of cognitive impairment. PMID- 16266830 TI - Desiderata for domain reference ontologies in biomedicine. AB - Domain reference ontologies represent knowledge about a particular part of the world in a way that is independent from specific objectives, through a theory of the domain. An example of reference ontology in biomedical informatics is the Foundational Model of Anatomy (FMA), an ontology of anatomy that covers the entire range of macroscopic, microscopic, and subcellular anatomy. The purpose of this paper is to explore how two domain reference ontologies--the FMA and the Chemical Entities of Biological Interest (ChEBI) ontology, can be used (i) to align existing terminologies, (ii) to infer new knowledge in ontologies of more complex entities, and (iii) to manage and help reasoning about individual data. We analyze those kinds of usages of these two domain reference ontologies and suggest desiderata for reference ontologies in biomedicine. While a number of groups and communities have investigated general requirements for ontology design and desiderata for controlled medical vocabularies, we are focusing on application purposes. We suggest five desirable characteristics for reference ontologies: good lexical coverage, good coverage in terms of relations, compatibility with standards, modularity, and ability to represent variation in reality. PMID- 16266831 TI - Transient isolation of Burkholderia multivorans and Burkholderia cenocepacia from a Brazilian cystic fibrosis patient chronically colonized with Burkholderia vietnamiensis. AB - Fifteen serial Burkholderia cepacia complex isolates recovered over a period of 4 years from a single cystic fibrosis patient were analysed for genomovar status by means of recA sequence determination, and genetic relatedness by RAPD-PCR. Twelve isolates were assigned as Burkholderia vietnamiensis, two as Burkholderia cenocepacia and one as Burkholderia multivorans. B. vietnamiensis persisted in the airways during 4 years, except in three occasions when B. cenocepacia or B. multivorans were isolated. The patient was chronically colonized by B. vietnamiensis with the RAPD-profile 12 and transiently by the RAPD-profile 15. PMID- 16266832 TI - Delayed diagnosis of cystic fibrosis associated with R117H on a background of 7T polythymidine tract at intron 8. AB - We report late diagnoses of cystic fibrosis (CF) in two men aged 61 and 65 years. At the time of presentation, both patients had significant pulmonary disease. In each case two CFTR gene mutations were identified, including R117H on a background of a poly T genotype of 7T/9T. Patients with two identified CFTR mutations which include the R117H/7T anomaly should be followed up routinely as they remain susceptible to severe lung disease. PMID- 16266834 TI - Purification and characterization of phycocyanin from the blue-green alga Aphanizomenon flos-aquae. AB - Aphanizomenon flos-aquae (AFA) is a blue-green alga and represents a nutrient dense food source. In this study the presence of phycocyanin (PC), a blue protein belonging to the photosynthetic apparatus, has been demonstrated in AFA. An efficient method for its separation has been set up: PC can be purified by a simple single step chromatographic run using a hydroxyapatite column (ratio A620/A280 of 4.78), allowing its usage for health-enhancing properties while eliminating other aspecific algal components. Proteomic investigation and HPLC analysis of purified AFA phycobilisomes revealed that, contrary to the well characterized Synechocystis and Spirulina spp., only one type of biliprotein is present in phycobilisomes: phycocyanins with no allo-phycocyanins. Two subunit polypeptides of PC were also separated: the beta subunit containing two bilins as chromophore and the alpha subunit containing only one. PMID- 16266833 TI - High-performance liquid chromatographic determination of acetyl-11-keto-alpha boswellic acid, a novel pentacyclic triterpenoid, in plasma using a fluorinated stationary phase and photodiode array detection: application in pharmacokinetic studies. AB - A rapid, sensitive and selective HPLC separation with photodiode array detection was developed for the analysis of the novel pentacyclic triterpenoid acetyl-11 keto-alpha-boswellic acid. Complete baseline separation of acetyl-11-keto-alpha boswellic acid from the corresponding isomer acetyl-11-keto-beta-boswellic acid was achieved on a fluorinated stationary phase. The standard curve was linear from 0.98 nmol/l to 196 nmol/l acetyl-11-keto-alpha-boswellic acid. The compound was isolated from chick embryonic plasma using extraction on diatomaceous earth with an overall average extraction yield of 82%. This method was applied in a kinetic study on the chick chorioallantoic membrane model (CAM) and showed unequivocal separation between acetyl-11-keto-alpha-boswellic acid and acetyl-11 keto-beta-boswellic acid unachievable so far. PMID- 16266835 TI - Solution 1H NMR investigation of Zn2+ and Cd2+ binding to amyloid-beta peptide (Abeta) of Alzheimer's disease. AB - Elevated levels of zinc2+ and copper2+ are found chelated to the amyloid-beta peptide (Abeta) in isolated senile plaque cores of Alzheimer's disease (AD) patients. However, the precise residues involved in Zn2+ ligation are yet to be established. We have used 1H NMR and CD to probe the binding of Zn2+ to Abeta(1 28). Zinc binding to Abeta causes a number of 1H NMR resonances to exhibit intermediate exchange broadening upon Zn2+ addition, signals in slow and fast exchange are also observed. In addition, there is a general loss of signal for all resonances with Zn2+ addition, suggestive of the formation of high molecular weight polymeric species. Perturbations in specific 1H NMR resonances between residues 6 and 14, and analysis of various Abeta analogues in which each of the three His residues have been replaced by alanine, indicates that His6, His13 and His14 residues are implicated in Zn-Abeta binding. Complementary studies with Cd2+ ions cause perturbations to 1H NMR spectra that are strikingly similar to that observed for Zn2+. Binding monitored at Val12 indicates a 1:1 stoichiometry with Abeta for both Zn2+ and Cd2+ ions. Circular Dichroism (CD) studies in the far-UV indicate quite minimal ordering of the main-chain with Zn2+ or Cd2+ addition. Changes in the far-UV are quite different from that obtained with Cu2+ additions indicating that Zn2+ coordination is distinct from that of Cu2+ ions. Taken together, these observations seem to suggest that Zn2+ coordination is dominated by inter-molecular coordination and the formation of polymeric species. PMID- 16266836 TI - Liver resection of metastasis by colorectal cancer in a HIV patient. PMID- 16266837 TI - Wireless capsule enteroscopy (Given) in a case of Cowden syndrome. PMID- 16266838 TI - Stable low levels of hepatitis C virus infection among Italian young males over the past decade. PMID- 16266839 TI - Life saving cyclophosphamide treatment in a girl with giant cell hepatitis and autoimmune haemolytic anaemia: case report and up-to-date on therapeutical options. AB - We report the case of a girl affected by giant cell hepatitis associated with autoimmune haemolytic anaemia. Both conditions were severe with a number of life threatening episodes of liver failure and anaemia unresponsive to several immunosuppressant drugs but cyclophosphamide. After a low-dose long-term treatment with this drug the patient is stably well without any therapy. A review of therapeutical options in this condition is also presented. PMID- 16266840 TI - Vaccine development in the 21st century: a time of living dangerously. PMID- 16266841 TI - Mycoplasmas - Identifying hosts for a stealth pathogen. PMID- 16266842 TI - The role of flow cytometry in companion animal diagnostic medicine. AB - Flow cytometry is a powerful tool for characterising the composition of complex cell populations. The accuracy and precision of this technology for describing and enumerating cells exceeds traditional methods. The number of diagnostic veterinary laboratories with access to a dedicated machine is increasing, and there is the potential to offer a clinical flow cytometry service. The improved availability of monoclonal antibodies (mAb) to cell markers expressed by the leukocytes of companion animals, permits the implementation of comprehensive mAb panels suitable for diagnosis of lympho- and myeloproliferative disease. Reticulated erythrocyte and platelet quantification, antiglobulin assays for immune-mediated cytopenias, lymphocyte subset analysis, and immunophenotyping of lymphoma and leukemia, have been validated for companion animal samples on the flow cytometer. It is now timely to consider the role of flow cytometry in diagnostic practice, and the requirement for quality assurance and standardization of testing procedures. PMID- 16266843 TI - Recent developments in veterinary vaccinology. AB - Advancement in technology and science and our detailed knowledge of immunology, molecular biology, microbiology, and biochemistry among other basic science disciplines have defined new directions for vaccine development strategies. The applicability of genetic engineering and proteomics along with other new technologies have played pivotal roles in introducing novel ideas in vaccinology, and resulted in developing new vaccines and improving the quality of existing ones. Subunit vaccines, recombinant vaccines, DNA vaccines and vectored vaccines are rapidly gaining scientific and public acceptance as the new generation of vaccines and are seriously considered as alternatives to current conventional vaccines. The present review focuses on recent advances in veterinary vaccinology and addresses the effects and impact of modern microbiology, immunology, and molecular biology. PMID- 16266844 TI - Mycoplasma host specificity: fact or fiction? AB - Bacteria of the genus Mycoplasma are the smallest organisms known to be capable of self-replication. They only occur in association with animal host cells on which they are dependant for many pre-formed nutrients since they lack many of the metabolic pathways associated with energy production and the synthesis of cell components found in other species of bacteria. It is generally thought that most species of Mycoplasma are very host specific but there are many reports of mycoplasmas in hosts that are not perceived as their normal habitat. Sometimes these "crossings" may have a pathological impact particularly where there may be predisposing conditions such as immunodeficiency. These are often reported in humans but may also occur in animals whose immune or physiological status is not known. This review brings together some of these reported incidents and speculates on their potential impact for laboratory diagnosis. PMID- 16266845 TI - Control of reproductive processes by growth hormone: extra- and intracellular mechanisms. AB - Recent data on the association between growth hormone (GH) and male and female reproductive processes, as well as the effects of GH on these processes and on some reproductive and non-reproductive disorders, and possible extra- and intracellular mediators of its action are reviewed. The available data suggest that GH is an important endocrine and autocrine/paracrine regulator of reproduction. It controls proliferation, apoptosis, growth and differentiation and the secretory and generative activities of different reproductive organs. It also regulates their response to gonadotrophin-releasing hormone (GnRH) and gonadotropins. Despite the effects of GH on the IGF/IGFBP (insulin-like growth factor binding protein) system, oxytocin, steroids, activin, gonadotropin and gonadotropin receptors, the majority of GH's actions on the reproductive processes are probably mediated not by these substances but by specific GH receptors acting through cAMP/protein kinase A, protein kinase G, tyrosine kinase , MAP kinase and CDC2 kinase-dependent intracellular mechanisms. Although GH treatments can increase the risk of some reproductive and non-reproductive disorders, they may be useful in improving gonadal function, inducing superovulation and in embryo production. PMID- 16266846 TI - Intracellular distribution of copper and zinc in the liver of copper-exposed cattle from northwest Spain. AB - The intracellular distribution of copper (Cu) and zinc (Zn) in the livers of normal and moderately Cu-exposed cattle was investigated with the aim of improving understanding of the pathophysiology of Cu accumulation in cattle. At total liver Cu concentrations within the generally accepted normal range (25-100 mg/kg fresh weight) the large-granule fraction was the main subcellular compartment for Cu accumulation, followed by the cytosol and the nucleus, whereas in the microsomal fraction Cu accumulation was very low. With increasing Cu exposure, the capacity of the large-granule fraction to accumulate Cu decreased, proceeding towards a plateau (estimated at about 80 mg/kg at a projected total liver Cu concentrations of 450 mg/kg), accompanied by progressively greater Cu accumulation in the nucleus and cytosol. Total liver Cu concentration had little influence on subcellular Zn distribution, with hepatic Cu concentrations being only moderately correlated with Zn concentration in the large-granule fraction. There was a strong association between the proportion of total Cu bound to metallothionein (MT) and the proportion of total Zn in the large-granule fraction, suggesting that Zn displaced from MT is taken up by the lysosomes for excretion. This pattern of Cu accumulation, as in sheep, may be due to the limited capacity for metallothionein binding of Cu and excretion in bile. PMID- 16266847 TI - Clindamycin bioavailability and pharmacokinetics following oral administration of clindamycin hydrochloride capsules in dogs. AB - Oral bioavailability and pharmacokinetic behaviour of clindamycin in dogs was investigated following intravenous (IV) and oral (capsules) administration of clindamycin hydrochloride, at the dose of 11 mg/kg BW. The absorption after oral administration was fast, with a mean absorption time (MAT) of 0.87+/-0.40 h, and bioavailability was 72.55+/-9.86%. Total clearance (CL) of clindamycin was low, after both IV and oral administration (0.503+/-0.095 vs. 0.458+/-0.087 L/h/kg). Volume of distribution at steady-state (IV) was 2.48+/-0.48 L/kg, indicating a wide distribution of clindamycin in body fluids and tissues. Elimination half lives were similar for both routes of administration (4.37+/-1.20 h for IV, vs. 4.37+/-0.73 h for oral). Serum clindamycin concentrations following administration of capsules remained above the MICs of very susceptible microorganisms (0.04-0.5 microg/mL) for 12 or 10 h, respectively. Time above the mean inhibitory concentration (MIC) is considered as the index predicting the efficacy of clindamycin (T(>MIC) must be at least 40-50% of the dosing interval), so a once-daily oral administration of 11 mg/kg BW of clindamycin can be considered therapeutically effective. For less susceptible bacteria (with MICs of 0.5-2 microg/mL) the same dose should be given but twice daily. PMID- 16266848 TI - Clostridium perfringens toxin-types in lambs and kids affected with gastroenteric pathologies in Italy. AB - A study was carried out in the South of Italy to assess the role of clostridia in neonatal diseases of lambs and kids. Eighty-seven lambs and 15 kids belonging to 25 flocks were examined and Clostridium perfringens was the microorganism most commonly identified. C. perfringens isolates were analysed by polymerase chain reaction (PCR), in order to determine the prevalence of the genes cpa, cpb, cpb2, etx, iap and cpe. The most prevalent toxin-type of C. perfringens was found to be type A found in 84% of the cases with clostridial enterotoxaemia. No C. perfringens type B, C or E were found. C. perfringens type D was isolated in 16% of the cases. About 24% of the isolates were cpb2 positive. The prevalence of cpb2 across the different C. perfringens types varied. The beta(2)-toxin gene cpb2 was detected in 4/21 (19%) type A isolates, in 1/2 type D isolates, and in 1/2 type DE (cpe-carrying type D) isolates. The high rate of positivity to cpb2 among the isolates suggests that a vaccine based on the beta(2)-toxin, should be included in the vaccination schedule of the animals to confer adequate protection and to prevent the disease. PMID- 16266849 TI - Morphology of caprine skin glands involved in buck odour production. AB - The distribution and morphology of the cornual, sub-caudal, mental and preputial glands were studied macro- and microscopically in four Toggenburg and eight miniature male goats. Although the cornual and sub-caudal glands could be readily located macroscopically, the mental glands in the inter-mandibular region and the preputial glands at the preputial orifice were not visible macroscopically. On histological section, all glands were found to be composed of lobulated sebaceous tissue combining both normal and modified holocrine secretory units. Over a period of 18 months, five consecutive glandular swabs for scent tests were taken to assess the influence of age and season on buck odour production. Buck odour was most apparent in the cornual gland area, less distinct at the mental gland region, and faint or absent in the other glandular areas. Surgical removal of the cornual glands caused a decrease in buck odour and persisting scent was ascribed to smaller skin glands dispersed in the cranial body half. Complete absence of buck odour was only observed in castrated bucks. PMID- 16266850 TI - Aerobic bacterial flora of the nasal cavity in Gulf of California sea lion (Zalophus californianus) pups. AB - Nasal swab samples from clinically healthy California sea lions pups (Zalophus californianus) from six different reproductive rookeries in the Gulf of California were collected to determine the type and frequency of the representative aerobic bacterial microflora of their nasal mucosa. A total of 114 samples were examined and 100 bacterial isolates were identified and typified by microbiological and biochemical standard tests. Fifty four isolates corresponded to Gram positive bacteria (54%) and 46 isolates to Gram negative bacteria (46%). Fifteen bacterial genera were identified, including Micrococcus, Arcanobacterium, Corynebacterium, Moraxella, Neisseria, Escherichia, Kurthia, Acinetobacter, Staphylococcus, Brevibacillus, Bacillus, Klebsiella, Stenotrophomonas, Pseudomonas and Aeromonas. The most frequently isolated genera were Moraxella (24%), Micrococcus (18%), and Corynebacterium (15%). These results show the presence in the nasal cavity of sea lions of several microorganisms. Although considered part of the normal microflora, they may also be opportunistic pathogens for their hosts and may act as a potential natural sentinel of environmental changes. PMID- 16266851 TI - Pharmacokinetics of marbofloxacin after intravenous and intramuscular administration to ostriches. AB - The pharmacokinetics of marbofloxacin was investigated after intravenous (IV) and intramuscular (IM) administration, both at a dose rate of 5 mg/kg BW, in six clinically healthy domestic ostriches. Plasma concentrations of marbofloxacin was determined by a HPLC/UV method. The high volume of distribution (3.22+/-0.98 L/kg) suggests good tissue penetration. Marbofloxacin presented a high clearance value (2.19+/-0.27 L/kgh), explaining the low AUC values (2.32+/-0.30 microgh/mL and 2.25+/-0.70 microgh/mL, after IV and IM administration, respectively) and a short half life and mean residence time (t(1/2 beta)=1.47+/-0.31 h and 1.96+/ 0.35 h; MRT=1.46+/-0.02 h and 2.11+/-0.30 h, IV and IM, respectively). The absorption of marbofloxacin after IM administration was rapid and complete (C(max)=1.13+/-0.29 microg/mL; T(max)=0.36+/-0.071 h; MAT=0.66+/-0.22 h and F (%)=95.03+/-16.89). PMID- 16266852 TI - Fractures sustained by racehorses in Japan during flat racing with special reference to track condition and racing time. AB - The purpose of this report is to describe the results of epidemiological surveys of racing-related fractures in Thoroughbred horses in Japan. In the period 1987 2000, a total of 10,203 fractures were diagnosed in 556,705 runners, resulting in an overall incidence of 1.83%. The annual incidence of fractures in flat racing during the 14-year period fluctuated between 1.44% and 2.19%. The majority of fractures affected the forelimbs. We found significant effects of track condition on injury incidence. The incidence of fractures decreased as track conditions on turf became softer and increased as track conditions on dirt became muddier. Because of the general trend for the incidence of fractures to vary with track condition, we examined the relationship between track conditions and racing times for winning horses in 4117 races and for 50,564 overall runners for the period 1990-1994 on two tracks. For turf courses, racing times became longer as track conditions became softer. In contrast, for dirt courses, racing time tended to become shorter in muddier conditions. The variation in the incidence of fracture with track condition might be due to the corresponding difference in racing time. PMID- 16266854 TI - Helicobacter spp. from captive bottlenose dolphins (Tursiops spp.) and polar bears (Ursus maritimus). AB - The gastric fluid of six bottlenose dolphins and the faeces of four polar bears from the same oceanarium were examined for the presence of Helicobacter. As detected by PCR, all dolphins and 8/12 samples collected from polar bears were positive for Helicobacter. Novel sequence types were identified in samples collected from these animals of which several were unique to either the dolphins or the polar bears. At least one sequence type was, however, detected in both animal taxa. In addition, a sequence type from a dolphin shared a 98.2-100% identity to sequences from other Helicobacter species from harp seals, sea otters and sea lions. This study reports on the occurrence of novel Helicobacter sequence types in polar bears and dolphins and demonstrates the broad-host range of some species within these animals. PMID- 16266853 TI - Anomalous insertion of the splenius muscle in a dog. AB - The splenius muscle in the dog is a triangular muscle extending from the third thoracic vertebra to the skull. It inserts on the nuchal line of the occipital bone and the mastoid part of the temporal bone and occasionally it also sends a strong serration to the transverse process of the axis. Here we describe a new, anomalous insertion of both the left and right splenius muscles in one young adult dog routinely prepared for anatomical dissection. In this dog, a group of muscle fibres diverged ventrolaterally from both the right and left splenial muscles to be inserted at the end of the transverse processes of the third cervical vertebra. This anomalous insertion of the splenius muscle corresponds to the splenius cervicis muscle present in other species. PMID- 16266855 TI - Clinical and pathological findings in a goat with cerebral gliomatosis. AB - This report describes a buck with cerebral gliomatosis. The animal was severely apathetic to somnolent. Neurological examination revealed generalised ataxia and hyper-metria of the fore limbs. There was bilateral mydriasis and severely decreased menace and pupillary light reflexes. Sensitivity to pricking with a needle was markedly reduced over the entire body. There was a delayed response to adduction, abduction and crossing of the limbs and rocking of the animal. Examination of cerebrospinal fluid indicated mild mixed-cell inflammation. Based on all of the findings, an abscess or tumour of the central nervous system with localisation in the cerebrum was suspected. Because of the grave prognosis, the goat was euthanased and a post mortem examination performed. No macroscopic abnormalities were seen in any of the organs including the brain. Histologically, there was extensive diffuse glial cell hyperplasia in the white matter of the cerebral hemispheres and in the brain stem. PMID- 16266857 TI - Regulation of interleukin-18 by THP-1 monocytoid cells stimulated with HIV-1 and Nef viral protein. AB - Interleukin (IL)-18 is a proinflammatory cytokine that plays an important role in both innate and adaptive immune responses against several infectious pathogens. Relatively little is known about its production in HIV-1 infection, and there are controversial data on the influence of IL-18 on HIV-1 replication in vitro. In this study, we investigated the effect of HIV-1 infection, and challenge with recombinant HIV-1 proteins, on IL-18 production by THP-1 cells. This is a monocytoid cell line spontaneously producing IL-18, and consequently is particularly suitable for the study of HIV-1 effects on this type of cytokine regulation. The results reported here demonstrate a significant reduction in IL 18 secretion during HIV-infection. In fact, low levels of IL-18 were released until 120 h from viral challenge (15 +/- 11 pg/mL at 24 h and 17 +/- 13 at 96 h and < 12.5 at 120 h), whereas IL-18 production by uninfected control cells was 193 +/- 104 pg/mL and 214 +/- 114 pg/mL at 24 h and 120 h respectively. At 168 h of incubation, IL-18 production by infected and uninfected cells was found to be 164 +/- 88 pg/mL and 325 +/- 101 pg/mL respectively (p = 0.001). Of the following viral proteins: gp 120, p24 and Nef, only the last one induced decreased IL-18 secretion in the supernatants of THP-1 cells. This effect is more evident with the concentrations of 5 -1.25 microg/mL of Nef protein (p < 0.0001). In conclusion, our data show that HIV-1 and its regulatory protein, Nef, are able to down-regulate the release of IL-18, in vitro. These results confirm that a variety of modulating effects on the immune response, induced by HIV-infection, may facilitate progression of HIV-1 infection. PMID- 16266856 TI - High serum interleukin-18 concentrations in patients with coronary artery disease and type 2 diabetes mellitus. AB - AIMS: The aim of our study was to analyse the serum level of interleukin 18 (IL 18) in coronary artery disease (CAD) patients with type 2 diabetes mellitus (DM), and to relate this to clinical findings. METHODS: The IL-18 level was measured by ELISA in serum samples from 130 CAD patients prior to their first, elective, coronary artery bypass surgery. Forty-three of them had been diabetic for several years. A control group consisted of 31 healthy people matched according to age, BMI, lipid and smoking status. RESULTS: The CAD patients with DM were similar to the non-diabetic CAD patients with respect to age, BMI, grade of heart failure, ejection fraction. There were no differences in the duration of CAD, history of myocardial infarction and PTCA or instability of angina. The serum level of IL-18 was higher in the CAD patients than in the control group. The CAD patients with DM had a higher concentration of IL-18 compared to the non-diabetic CAD group. The diabetic patients with triple-vessel disease were characterized by a higher concentration of IL-18 than the non-diabetic patients with the same grade of CAD. Smoking affected the IL-18 concentration, particularly in the diabetic patients. CONCLUSION: Type 2 DM predisposes patients, especially those with multi-vessel CAD who were smokers, to a higher serum level of IL-18, which may help explain their vulnerability to fatal, secondary cardiovascular events. These patients should be in the first line for stringent, secondary cardiovascular prevention. PMID- 16266858 TI - IFN-gamma is not induced through increased plasma concentrations of interleukin 12/interleukin-18 during human endotoxemia. AB - Endotoxin administration to animals and humans is an accepted experimental model of Gram-negative sepsis, and endotoxin is believed to play a major role in triggering the activation of cytokines. In septic patients, the IL-12/IL-18/IFN gamma axis is activated and correlates with mortality. Our aim was to investigate the effects of endotoxin administration in humans on the activation of the IL 12/IL-18/IFN-gamma axis. Seven healthy volunteers received E. coli endotoxin (O:113). Hemodynamics, temperature and the course of plasma concentrations of TNF alpha, IL-1beta, IL-12, IL-18 and IFN-gamma were determined. Endotoxin administration resulted in the expected flu-like symptoms, a temperature of 38.8 +/- 0.3(o)C (p < 0.003), a decrease in mean arterial blood pressure of 14.8 +/- 1.8 mmHg (p < 0.0002) and an increase in heart rate of 27.5 +/- 4.8 bpm (p < 0.002) compared to baseline values. TNF-alpha increased from 16.6 +/- 8.2 to 927 +/- 187 pg/mL (p < 0.003). IL-1beta increased from 8.6 +/- 0.5 to 25.3 +/- 2.0 pg/mL (p < 0.0001). IL-12 showed no significant increase (8.2 +/- 0.2 to 9.3 +/- 0.8 pg/mL, p = 0.13), and all IL-18 measurements remained below the level of detection. In contrast, IFN-gamma showed an increase from 106.6 +/- 57.1 to 152.7 +/- 57.8 (p < 0.005). These results indicate that pathways other than the IL 12/IL-18 axis may induce IFN-gamma production in human endotoxemia. PMID- 16266859 TI - The effect of fish oil supplementation on cytokine production in children. AB - The ex vivo production of inflammatory cytokines during fish oil supplementation (n-3 polyunsaturated fatty acids, n-3 PUFA) is a matter of considerable controversy. Studies on human subjects have generally reported decreased lymphocyte proliferation and decreased production of IL-2, interferon-gamma, IL 1beta, IL-6 and TNF-alpha, but other studies showed no effect or even increased production. There are no published reports on ex vivo cytokine production in children on long-term, n-3 PUFA supplementation. The current double-blind study explored cytokine production by peripheral blood mononuclear cells (PBMCs), with and without lipopolysaccharide (LPS) stimulation in children on 12 weeks' supplementation with 300 mg/day of n-3 PUFA. Twenty-one children (aged 8-12 years) were randomized to receive 1 g canola oil (control) or 300 mg n-3 PUFA + 700 mg canola oil in a chocolate spread. Blood was then drawn and PBMCs were separated and cultured for 24 h in a culture medium with or without 10 microg/mL LPS for 5 x 10(6) PBMCs. The pro-inflammatory cytokines, IL-1beta, TNF-alpha and IL-6, and the anti-inflammatory cytokines, IL-10 and IL-1RA, were evaluated by ELISA. The levels of all the cytokines were higher in non-stimulated and LPS stimulated cultures, from n-3 PUFA-treated subjects as compared to controls. There was no difference in the IL-1beta/IL-1RA ratio between the two groups, with and without LPS stimulation. Nevertheless, the ratio tended to be lower in the treated subjects on both occasions. In conclusion, our results indicate an increased production of both pro-inflammatory and anti-inflammatory cytokines, with and without LPS stimulation, in children on 12 weeks' n-3 PUFA supplementation. PMID- 16266860 TI - Comparison of serum levels of seven cytokines in premature newborns undergoing different ventilatory procedures: high frequency oscillatory ventilation or synchronized intermittent mandatory ventilation. AB - OBJECTIVE: The severity of pulmonary dysfunction and subsequent development of chronic lung disease (CLD) in preterm neonates depends on several factors, among them oxygen administration. The aim of this report is to compare the effects of high-frequency, oscillatory ventilation (HFOV) versus synchronized, intermittent, mandatory ventilation (sIMV) on serum cytokine levels (IL-6, IL-8, IL-10, MCP-1, PDGF-BB, VEGF and TGF-beta1) and ventilator indices during the first week of life. Moreover, CLD development and several other outcomes were compared between the two groups. DESIGN: Randomized clinical trial. SETTING: Third level NICU. PATIENTS: 40 preterm neonates with a gestational age between 24 and 29 weeks were randomly (20 per group) assigned to one of the two, above-mentioned ventilation strategies within 30 minutes of birth. MEASUREMENTS AND RESULTS: At 1, 3 and 5 days, neonates were monitored by means of ventilator indices and levels of seven pro-inflammatory or anti-inflammatory (pro-fibrotic) cytokines in serum. No clinical or biochemical differences were observed at baseline. The neonates assigned to HFOV benefited from early and sustained improvement in gas exchange, with earlier extubation and lower incidence of CLD, as compared to the neonates assigned to sIMV treatment, and showed a significant reduction of serum IL-6, IL 8 and IL-10 over time only when the HFOV treatment was administered. In addition, at days 3 and 5, the IL-6 levels were significantly lower in the HFOV group as compared to sIMV patients. CONCLUSIONS: The results of this randomized clinical trial support the hypothesis that early use of HFOV, combined with an optimum volume strategy, has a beneficial effect, reducing serum levels of pro inflammatory cytokines and consequently the acute phase leading to lung injury. PMID- 16266861 TI - Breastfeeding reduces immune activation in primary respiratory syncytial virus infection. AB - In epidemiological studies of respiratory syncytial virus (RSV) disease, breast milk has proven to be beneficial. However, a host mechanism that is associated with both disease severity and that is capable of being modulated by breast milk, has not yet been identified. Both the predominance of interleukin-10 (IL-10) over interferon-gamma (IFN-gamma), and high soluble interleukin-2 receptor antagonist (sCD25) concentrations have been associated with RSV severity. We explored if they were modulated by breastfeeding. Previously healthy Chilean infants from Santiago with RSV infection (n = 349) were consecutively enrolled in the study if they were term births, without underlying pathology. Breastfeeding was described as absent or present, and if partial or exclusive. Immune response was expressed through plasma concentrations of IFN-gamma, IL-10 and sCD25, obtained both in the acute and the recovery phase. The acute phase sCD25 concentrations were lower in the breastfed (13.8 ng/mL, n =133), compared with the non-breastfed infants (15.9 ng/mL, n 27, p = 0.015). The difference increased in infants below 3 months of age (p = 0.006) and with exclusive (p = 0.004), compared to partial breastfeeding (p = 0.025). When analyzed together with age, sex, severity and environment, breastfeeding was the only independent predictor of high sCD25 concentrations (above mean + 1SD, OR 4.6, 95% CI 1.8-11.9, p = 0.0015). The recovery phase IFN gamma/IL-10 ratio was higher in the breastfed infants, but when analyzed with potential confounding factors, only female sex was associated with an increased ratio (OR 2.32, 95% CI 1.02-5.29, p = 0.045). High sCD25 concentrations during the acute phase of infection, previously associated with severe RSV disease, were significantly and independently reduced in association with breastfeeding, whereas the Th1/Th3 balance was only modified in the recovery phase. PMID- 16266862 TI - Changes in metalloproteinases in healthy normotensive patients with high-normal blood pressure. AB - INTRODUCTION: High-normal blood pressure (HNBP) seems to be related to increased cardiovascular risk in healthy, normotensive subjects, while essential hypertension is associated with an increase in extracellular matrix content, especially fibrillar collagen type I. The aim of our study was to investigate whether collagen degradation is altered in healthy normotensives with HNBP, and whether this alteration could be related to disturbances in the matrix metalloproteinases plasma concentration, and to compare the findings to those of healthy normotensives with normal blood pressure (NBP) levels, matched for age, sex and BMI. METHODS: Twenty six (14 males, 12 females) healthy, normotensive patients with HNBP, mean age 52 +/- 5 yrs, and BMI 23 +/- 1.5 kg/m(2) (group A), and 24, healthy normotensive patients (13 males, 11 females) with NBP, mean age 53 +/- 6 yrs, and BMI 23.2 +/- 1.4 kg/m(2) (group B), were studied. The two groups were matched for age, sex and BMI. Plasma levels of matrix metalloproteinase-9 (MMP-9) and tissue inhibitors (TIMP-1) and (TIMP-4) were determined by relevant ELISA in the study population. RESULTS: Plasma MMP-9 levels were significantly higher, while TIMP-1 and TIMP-4 levels were significantly lower in group A compared to group B, (MMP-9 579 +/- 147 versus 294 +/- 111 ng/mL, TIMP-1 178 +/- 45 versus 237 +/- 35 ng/mL p < 0.01, and TIMP-4 2.2 +/- 1.4 versus 4.4 +/- 2.1 p < 0.04 respectively). CONCLUSIONS: Our findings suggest that healthy normotensives with high-normal blood pressure have significantly increased MMP-9 and decreased TIMP-1 and TIMP-4 plasma levels compared to healthy normotensives with normal blood pressure. These findings need further investigation. PMID- 16266863 TI - Development of a sensitive ELISA for the quantification of human tumour necrosis factor-alpha using 4 polyclonal antibodies. AB - Despite the availability of many assays to measure concentrations of tumour necrosis factor alpha (TNF-alpha) in body fluids, these assays often lack specificity or sensitivity and are often of questionable reliability, resulting in inconsistent results. Therefore, we have developed an ELISA that is sensitive, reliable and not susceptible to disturbances by interfering substances such as heterophilic antibodies. The assay involves a combination of four polyclonal antibodies. The antibodies, which capture the analyte, were raised in chicken and the trapping anti-analyte antibodies were raised in rabbit. The immobilization of capture antibodies was achieved via a coating antibody raised in a duck against chicken IgY and the recognition of trapping antibodies was achieved by a detection antibody raised in a goat against rabbit IgG and labelled with HRP. The analytical and functional sensitivities of the ELISA are 8 pg/mL and 13 pg/mL, respectively. The assay showed good precision and, in contrast to our in-house RIA, excellent parallelism in serial dilutions. The recovery of TNF-alpha spiked to plasma samples ranged from 97% to 119%. Comparison of the newly developed, sensitive ELISA with our in-house RIA showed that the median TNF-alpha value obtained by RIA (range: 0.095-10.0, median 0.578 ng/mL) was found to be 1.5-2 times higher than that obtained with the ELISA (range 0.008-5.84, median 0.213 ng/mL). Spearman correlation was 0.755 (p < 0.0001). In addition, analysis of the TNF-alpha concentrations in blood from healthy individuals and from patients suffering from tuberculosis, with RIA and ELISA, showed the same differences although TNF-alpha levels obtained with ELISA were lower. We feel that this ELISA is a major improvement compared to the currently available assays for TNF. PMID- 16266864 TI - Spontaneous and cytokine-evoked production of matrix metalloproteinases by bone marrow and peripheral blood pre-B cells in childhood acute lymphoblastic leukaemia. AB - The present work focused on the study of the secretory activity of pre-B acute lymphoblastic leukaemia (ALL) cells harvested from bone marrow (BM) and peripheral blood (PB) in 16 children. The basal and cytokine (SDF-1, GM-CSF, bFGF, VEGF)-stimulated secretions of gelatinases 2 and 9 (MMPs-2 and -9) and expression of their genes were monitored by zymography and RT-PCR, respectively. A wide heterogeneity was found in the secretory capacities of these cells. The basal secretion of MMP-9 was more frequently observed than that of MMP-2 in both cell types. The cytokines VEGF and bFGF were found to induce predominant stimulatory effects on the MMP-2 secretion. In contrast, GM-CSF was shown to exert a more pronounced activation of the MMP-9 production. Experiments using inhibitors of metabolic pathways (U0126, LY294002 and SN50) revealed that the secretion of MMP-9 was mediated through PI3/MEK1 kinases. The MMP-2 secretion appeared to be however, stimulated through a different metabolic pathway. The microfluorimetric approach showed that the basal and stimulated secretions of MMPs-2 and -9 depended on the extracellular calcium pool. The cytokines VEGF and bFGF represent potent factors increasing the intracellular calcium concentration with similar kinetics. In contrast, GM-CSF was found to activate a verapamil sensitive efflux of indo-1 from cytosol suggesting that this cytokine could be responsible for the activation of xenobiotic membrane transporters. Experiments using the trypan blue exclusion test demonstrated that bFGF, in contrast to VEGF and GM-CSF, markedly augmented pre-B ALL cell survival. Further investigations into a possible correlation between the plasma concentrations of MMP-2 and -9, VEGF, bFGF and GM-CSF, and the poor evolution of pre-B ALL in children could have valuable diagnostic implications. PMID- 16266865 TI - Interleukin-9 stimulates the production of interleukin-5 in CD4+ T cells. AB - We recently showed that interleukin-9 (IL-9), a Th2 cytokine, promotes IL-5 mediated rejection of allografts in mice. This observation led us to investigate the functional link between IL-9 and IL-5 production during alloreactive T cell responses in vitro and in vivo. Firstly, we found that IL-9 was produced by alloreactive Th2 cells, and IL-9 mRNA was detected in skin allograft during Th2 type rejection. We then established that IL-5 production was impaired in alloreactive Th2 cells isolated from IL-9-deficient mice and that optimal IL-5 production after allogeneic stimulation requires a functional IL-9 receptor (IL 9R) on the responding cells. Finally, the production of IL-5 by anti-CD3 stimulated CD4+ T cells was abolished by neutralization of IL-9. Despite the fact that IL-9 promotes IL-5 production by alloreactive T cells, IL-9-deficient recipients of skin allografts still developed eosinophilic graft infiltrates and neither IL-9 nor IL-9R deficiency modified Th2-type allograft rejection. PMID- 16266867 TI - Follicular lymphoma: a therapeutic update. AB - Follicular lymphoma is the most common low-grade non Hodgkin's lymphoma. This indolent disease is usually characterised by an indolent course with a continuous pattern of relapse and a median survival of 10 years. Radiotherapy can be curative in a small proportion of patients with very localized disease, but the majority of patients have advanced disease at diagnosis and it is not clear that any current therapy is curative in this situation. In the last 5 years we have seen a dramatic increase in the number of patients receiving emerging therapies such as antibody therapy combined or not with chemotherapy, radioimmunotherapy or stem cell transplantation. In this article, we will review the available treatments of follicular lymphomas, with special emphasis on published phase III trials. PMID- 16266866 TI - Inflammatory cytokine profile and circulating cortisol levels in malnourished children with necrotizing ulcerative gingivitis. AB - Necrotizing ulcerative gingivitis (NUG), a periodontal disease traditionally associated with stressful lifestyles in young adults in developed countries, is very prevalent in socioeconomically deprived Nigerian children. Random incident cases (153) of NUG, along with their neighborhood village counterparts of comparable age and without NUG, as control, were recruited for this study. Anthropometric evaluation revealed widespread malnutrition and poor health in both groups of children, with more severe stunting in NUG cases. The poor nutritional status of the village children, with and without NUG, was also confirmed by markedly reduced levels of circulating micronutrients. Compared with the neighborhood children, NUG victims showed significant (p < 0.05 or < 0.001) increases in serum levels of interleukin (IL)-8 (+ 233%), IL-18 (+ 30%), IL-6 (+ 190%), IL-1beta (+ 341%), IL-10 (+ 186%), with a small decrease in interferon (IFN)-gamma (-19%) and nonsignificant increases in soluble tumor necrosis factor (TNF) receptors (sTNFR-p55, p75). Associated with NUG was a significant, 38% (p < 0.05) increase in plasma cortisol above the already high levels observed in the neighborhood village children, as well as some micronutrient deficiencies. The findings suggest that NUG is associated with dysregulated cytokine production, with a complex interplay of elevated levels of pro- and anti-inflammatory mediators. Such changes may serve as the common link between the seemingly unrelated risk conditions (e.g. stressful life styles, smoking, microbial infections, diabetes, malnutrition, alcoholism) traditionally implicated in the genesis of NUG, and all known to promote an increase in the blood level of cortisol, as well as a Th(1) to Th(2) cytokine shift. PMID- 16266868 TI - [Negative regulation of the JAK/STAT: pathway implication in tumorigenesis]. AB - Cytokines, hormones or growth factors induce a variety of biological responses including proliferation, differentiation and apoptosis. After binding to their specific cell surface receptors, these stimuli induce the activation of a number of signaling pathways including the activation of JAK (JAnus Kinase) proteins by auto and transphosphorylation. Activated JAK phosphorylate the receptor chains on tyrosines, creating docking sites for cytoplasmic transcription factors named STAT (Signal Transducers and Activators of Transcription). Furthermore, the JAK phosphorylate the STAT which form dimers and migrate to the nucleus where they bind to specific DNA sequences leading to the activation of transcription. The multiplicity of JAK (4 members) and STAT (7 members) and their associations with multiple possible partners allow the formation of various STAT homo and heterodimers and STAT-containing transcriptional complexes. Each of these complexes lead to the specific regulation of gene transcription. Negative regulation of the JAK/STAT signaling pathway is crucial to switch off the cytokine/growth factors' signal. Three families of proteins : the phosphotyrosine phosphatases (SHPs, CD45, PTP1B/TC-PTP), the SOCS proteins (Suppressors Of Cytokine Signaling) and the PIAS (Protein Inhibitor of Activated STAT) are involved in this process. These proteins act at various levels of the JAK/STAT pathway. Thus, tyrosine-phosphatases dephosphorylate activated JAK, STAT or cytokine receptors. PIAS interact with activated STAT and inhibit their DNA binding or their transactivating capacity, probably in relation with their intrinsic SUMO E3-ligase activity. The tyrosine phosphatases and the PIAS are constitutively present in the cell and represent a first level of regulation. The SOCS, which represent a second level of JAK/STAT negative control, are induced by cytokines and exert a negative feed-back loop. Indeed, they interact with activated JAK or with phosphorylated receptors, inhibiting the recruitment of STAT, the activation of the JAK enzymatic activity, or inducing the proteasome dependant degradation of activated JAK or receptors. PMID- 16266869 TI - [Combined role of tumor markers and 18fluoro-deoxyglucose-positron emission tomography (18FDG-PET) in follow-up of cancer patients]. AB - One of the major indications of tumor markers is the detection of occult disease. Less than 20 % of tumor markers elevations are associated with clinical or radiological findings. Such elevations have led medical community to doubt about the interest of markers follow-up, such as CA 15.3 in breast cancer. Now, positron emission tomography with (18)fluoro-desoxyglucose (18FDG-PET), using metabolic properties of malignant cells, is able to visualize tumor recurrences at an early stage of development, before any occurrence morphologic changes depicted by radiological examinations. Because of its cost and its limited accessibility, this functional technique should only be prescribed following a large set of informative indications, of which tumor markers belong. Early positive, non invasive and cost effectiveness, tumor markers become a precious guide in the prescription of 18FDG-PET in oncology. This article reviews the results of a set of recent studies in colorectal, breast and ovarian cancer. PMID- 16266870 TI - [Prostate cancer: update]. AB - The study of mitotic transduction of the signal showed that overexpression of pAkt and reduction in pERK expression would be associated a biological relapse. For tumors T1-3 N0M0 at the high risk of local relapse after prostatectomy, an immediate radiotherapy compared with a differed radiotherapy (at the time of PSA relapse), showed a significant reduction in the rate of local relapse and an ameliorated progression free survival. The effectiveness of the docetaxel was confirmed in two phase III randomized clinical trials : TAX-327 with 3 arms compared docetaxel every 21 days, docetaxel every 7 days and mitoxantrone. All arms were prednisone-based. An increase in overall survival, PSA progression free survival, PSA response rate and a pain reduction were highlighted in the docetaxel arm every 21 days. Docetaxel obtained at the end of this study the marketing authorization in this indication and became the treatment of reference. The SWOG 99-16 study compared the docetaxel estramustine association with the same arm of reference, mitoxantrone and prednisone, with similar results. The addition of estramustine to the docetaxel seems to improve the PSA response rate and progression free survival, but with a greater embolic toxicity. The addition of an antiangiogenic agent, the thalidomide, to docetaxel, improves progression free survival and overall survival. PSA responses were observed with an inhibitor of the proteasome, the bortezomib, in monotherapy, contrary to the imatinib which in monotherapy didn't have any effectiveness. Studies in association with docetaxel are ongoing. Some biological responses were observed with a vaccine anti MUC-1 and must be confirmed on a greater series of patients. The docetaxel impact on localized disease is actually evaluated. PMID- 16266871 TI - [Breast cancer screening for women with a strong familial risk]. AB - Screening for breast cancer in high-risk women could be seen through the general criteria for cancer screening. The attributable part of BRCA in breast cancer is estimated to be between 2 and 5%. For these women, breast cancer screening lay between prevention with low risk/benefit efficacy and prophylactic mastectomy with high efficacy but low acceptability. Risk reduction could be achieve with "classic" screening tools (examination, mammography and ultra sound) but should soon benefit from MRI experimental protocols. Later, combined imaging and biology protocols may solve the issue of screening in cancer prone women. A comprehensive program is required, taking in account both the competitive risk of ovarian cancer and the risk reduction observed at the breast level with prophylactic oophorectomy, at least in BRCA1 women. Information, and psychological help is all the more a necessity, since the lack of specificity of screening may induce harm, particularly for these women. Due to methodological flaws, the low power and a short follow-up of the surveys, this statement cannot however aspire to a high stability. PMID- 16266872 TI - [Advantage of immunochemical fecal occult blood test in screening for colorectal cancer]. AB - Colorectal cancer screening is a high public health priority in all industrialized countries. However, the low sensitivity of the common guaiac screening test (Haemoccult II) makes practitioners and public health deciders reluctant to set up national screening program. In recent years, immunochemical tests based on the use of a specific antibody have been found to be more sensitive than the Haemoccult II test. However, for screening purposes, any gain in sensitivity is of interest only if specificity and positive predictive value are satisfactory. As instance, rehydration of the Haemoccult II test prior to lecture can increase sensitivity, but the associated decrease in specificity and positivity predictive value and the high positivity rate render its value in mass screening debatable. Moreover, extra costs, if existing, must be acceptable for the society. Until recently, immunochemical tests costs made it unaffordable in our societies. The arrival of automated reading is likely to remove this obstacle. Moreover it offers the opportunity of positivity cut-off choice. A recent study was conducted in Cotentin (France) to assess the performance of an immunochemical test with an automated reading technique (Magstream 1000) for different haemoglobin content cut-off points. As previous American, Japanese, Chinese and Italian studies, this study suggests that the use of immunochemical tests could lead a substantial gain, in screening sensitivity. Moreover by choosing a higher haemoglobin content as cut-off point (50 ng/ml instead of usual cut-off at 20 ng/ml), a gain in sensitivity can be obtained with a satisfactory specificity (97%) and positivity rate (3%). Considering the increasing number of publications, the use of an immunochemical test with an automated reading technique could improve the prospects for mass-screening for colorectal cancer, since it offers a promising alternative to guaiac tests. PMID- 16266873 TI - [Standards and Options for the use of radiation therapy in the management of patients with osteosarcoma. Update 2004]. AB - CONTEXT: The A "Standards, Options and Recommendations A" (SOR) project, started in 1993, is a collaboration between the Federation of French Cancer Centres (FNCLCC), the 20 French regional cancer centres, and specialists from French public universities, general hospitals and private clinics. The main objective is the development of clinical practice guidelines to improve the quality of health care and the outcome of cancer patients. The methodology is based on a literature review and critical appraisal by a multidisciplinary group of experts, with feedback from specialists in cancer care delivery. OBJECTIVE: To update the SOR recommendations for the use of radiation therapy in the management of patients with osteosarcoma. This work was performed in collaboration with the French society against cancers in children and adolescent (SFCE). METHODS: Data have been identified by literature search using MedlineA (from January 1992 to October 2003). In addition several Internet sites were searched in October 2003. RESULTS: The 3 mains standards are: 1) local and exclusive curative irradiation is not indicated as primary treatment for osteosarcoma or for local and operable recurrence, except for lesion in inaccessible sites or if the patient refuses surgery; 2) local and prophylactic adjuvant irradiation is not indicated for the treatment of osteosarcoma after chemotherapy (neoadjuvant and/or adjuvant) and complete macro or microscopic surgery, except for nonoperable R1 or R2 surgical resection and 3) whole-lung prophylactic irradiation is not indicated in non metastatic osteosarcoma. Systemic metabolic radiotherapy for pain treatment, using samarium-153 ethylene diamine tetramethylene phosphonic acid (Sm-153-EDTMP) can be offered to patients with painful metastatic osteosarcoma or in case of recurrent bone sites inaccessible to local therapies (surgery, external irradiation). PMID- 16266874 TI - [Recommendations for the management of GIST patients]. AB - BACKGROUND: The management of gastrointestinal stromal tumors (GIST) has evolved very rapidly in the last years. A national consensus meeting was therefore organized in order to identify the optimal management procedures for patients with GIST in localized and advanced stages. METHODS: A panel of different specialties, including pathology, molecular biology, imaging, surgery, gastroenterology, medical oncology reviewed the current literature, in particular the recent Lugano conference, to identify consensus points and topics for future research in four different working groups: pathology and molecular biology, early management of small tumors and imaging, surgery, and medical treatment. Consensus points were categorized according to the Standard Options Recommendations (SOR) of the French Federation of Cancer Centers. RESULTS: The standard histological examination with immunohistochemical analysis using CD117, CD34, PS100, desmin and smooth muscle actin is considered standard. Molecular biology for the identification of KIT and PDGFRA mutation is advisable for GIST with negative CD117 staining, and otherwise is considered a research procedure. Complete tumor resection with negative tumor margins is the standard surgical treatment. Adjuvant imatinib after optimal tumor resection as well as neo-adjuvant imatinib remain experimental approaches to be performed within prospective clinical studies. Imatinib should be started at the date of diagnosis of metastatic relapse and given until development of intolerance or progressive disease. Resection of metastases is also considered as an experimental procedure which can not be recommended routinely. The criteria for tumor response to imatinib should include not only tumor size reduction or disease stabilization, but also reduction of tumor density (Hounsfield units) on computed tomography, metabolic activity (i.e. reduction of FDG uptake on positron emission tomography), and reduction of vascularisation of the tumors using contrast enhanced ultrasound evaluation. An increase in tumor size may be associated with pathologic response to imatinib therapy, and available survival data indicate that the survival of these patients is similar to that of patients with conventional tumor response. CONCLUSIONS: Consensus points in clinical management of GIST in this national conference adopted the majority of consensus points published in the Lugano conference. This multidisciplinary work will be published in the reference oncology, gastroenterology, and pathology journals in French languages. PMID- 16266890 TI - Chronic lymphocytic leukemia--aiming at a moving target! PMID- 16266891 TI - Interactions between RPS19, mutated in Diamond-Blackfan anemia, and the PIM-1 oncoprotein. AB - BACKGROUND AND OBJECTIVES: Diamond Blackfan anemia (DBA) is a congenital disease characterized by defective erythroid progenitor maturation. Patients' bone marrow progenitor cells do not respond to erythropoietic growth factors, such as erythropoietin. Mutations in the gene encoding for ribosomal protein (RP) S19 account for 25% of cases of DBA. The link between defective erythropoiesis and RPS19 is still unclear. Two not mutually exclusive hypotheses have been proposed: altered protein synthesis and loss of unknown extraribosomal functions. DESIGN AND METHODS: We used yeast two-hybrid screening and a human liver cDNA library obtained at 19-24 weeks of gestation, when hepatic erythropoiesis is efficient, to search for proteins interacting with RPS19. RESULTS: We found that RPS19 binds PIM-1, an ubiquitous serine-threonine kinase whose expression can be induced in erythropoietic cells by several growth factors, such as erythropoietin. The PIM 1/RPS19 interaction was demonstrated both in vitro and in living cells and led to phosphorylation of RPS19 in an in vitro kinase assay. We also showed that in human 293T cells PIM-1 interacts with ribosomes and may be involved in translational control. Three DBA-associated RPS19 mutations alter the binding between RPS19 and PIM-1. INTERPRETATION AND CONCLUSIONS: A link between erythropoietic growth factor signaling and RPS19 has been identified for the first time. PMID- 16266892 TI - A novel 5' ATRX mutation with splicing consequences in acquired alpha thalassemia myelodysplastic syndrome. AB - BACKGROUND AND OBJECTIVES: Acquired alpha thalassemia (hemoglobin H (HbH) disease) is a rare complication of neoplastic chronic myeloid disorders, especially myelodysplastic syndrome. Acquired HbH has recently been associated with mutations in an X-linked gene, ATRX, previously linked to inherited ATR-X syndrome (alpha thalassemia-retardation-X linked). DESIGN AND METHODS: A Swiss man with chronic myelomonocytic leukemia complicated by various autoimmune disorders and by strikingly microcytic, hypochromic anemia was analyzed for the presence of acquired HbH. After HbH detection, we sought an underlying genetic cause. We used denaturing high-performance liquid chromatography to screen for an ATRX mutation, and measured ATRX expression by reverse transcriptase polymerase chain reaction. RESULTS: The patient had 50% HbH-containing cells on supravital staining. Marrow karyotype and the alpha globin cluster were normal. A clonally restricted ATRX point mutation was detected in the conserved splice donor motif in intron 4 (IVS 4 +2 T-->C). Plasmid vector cloning of patient ATRX cDNA demonstrated both exon 4 skipping and partial intron retention with activation of a cryptic splice site, both outcomes resulting in frameshifts with premature stop codon generation in exon 5 and near-decimation of ATRX expression in myeloid cells. Normal exon 6 alternative splicing was retained. INTERPRETATION AND CONCLUSIONS: Intronic ATRX mutations with splicing consequences, uncommon in inherited ATR-X syndrome because of their devastating effect on expression of functional protein, should be routinely sought when undertaking molecular analysis of acquired HbH disease. Detection of an acquired ATRX mutation can help support clonality in karyotypically normal ambiguous myeloid disorders with HbH. PMID- 16266893 TI - The impact of RNA stabilization on minimal residual disease assessment in chronic myeloid leukemia. AB - BACKGROUND AND OBJECTIVES: Accurate quantification of BCR-ABL mRNA is of critical importance for managing patients with chronic myeloid leukemia (CML) who are receiving imatinib therapy. RNA degradation thus constitutes a potential problem for laboratories quantifying minimal residual disease (MRD). Patients' samples that take a long time to be transported from the hospital to the analyzing laboratory may be subject to RNA degradation with a corresponding loss in sensitivity and possible generation of false negative results. Recently, RNA preservation systems have been developed in order to improve RNA stability. The aim of the present study was to investigate such a system. DESIGN AND METHODS: We evaluated the performance of the PAXgene Blood RNA Kit in follow-up CML peripheral blood samples and compared the results to those from unstabilized parallel Trizol extracted samples. The different sample processing methods were evaluated by real-time polymerase chain reaction (PCR) analysis. RESULTS: RNA isolated with the PAXgene system gave a superior yield per milliliter of blood than did the routine Trizol extraction method. However, although of comparable quality, the RNA did not PCR-amplify as efficiently as equal amounts of RNA from routinely processed samples. Therefore, RNA processed with the PAXgene system showed decreased sensitivity for MRD detection, resulting in false negative results. The sensitivity was comparable to that of samples processed routinely 20 30 hours after phlebotomy. INTERPRETATION AND CONCLUSIONS: We conclude that routinely processed, i.e. unstabilized, peripheral blood that reaches the laboratory and is processed within 30 hours is preferable for MRD detection. Optimal results were achieved with fresh samples processed within 5 hours with the Trizol method. However, RNA stabilization may be useful if sample transit is expected to exceed 30 hours. PMID- 16266894 TI - The management and outcome of 18 pregnancies in women with polycythemia vera. AB - BACKGROUND AND OBJECTIVES: Polycythemia vera (PV) is rare in women of childbearing age with only 20 previous pregnancies reported. DESIGN AND METHODS: We report a series of 18 pregnancies (19 fetuses) in eight women with PV managed prior to or following implementation of management guidelines tailored to PV in pregnancy, and review the literature. RESULTS: Seven of these pregnancies were managed by standard antenatal care (group A) without specific attention to the women's PV. All remaining 11 pregnancies (group B) were managed following a formal protocol and received tailored management principally comprising tight control of the hematocrit by venesection, and the use of interferon ? in three patients, in addition to aspirin 75 mg, and prophylactic low molecular weight heparin (LMWH). Each pregnancy was monitored with uterine artery Doppler examinations and regular fetal scanning. In group A (n=7) there was one live birth, which required delivery at 34 weeks due to placental insufficiency, three first trimester miscarriages, two stillbirths and one combined stillbirth and neonatal death (twins) associated with placental dysfunction. All 11 patients in group B received aspirin and post-partum LMWH; four also received venesection (during pregnancy), three interferon-a and three antenatal LMWH. There were ten live births, nine at term, one first trimester miscarriage and no intrauterine growth retardation. INTERPRETATION AND CONCLUSIONS: Pregnancy in PV without meticulous attention to hematocrit is associated with poor fetal outcome. Aggressive intervention with control of hematocrit, aspirin and some LMWH appears to be associated with significantly better outcome (p=0.0017). PMID- 16266895 TI - Gene-expression profiles and their association with drug resistance in adult acute myeloid leukemia. AB - BACKGROUND AND OBJECTIVES: From 20-50% of patients with acute myeloid leukemia (AML) are primarily resistant to induction chemotherapy. It has previously been shown that resistance to the first cycle of induction chemotherapy is an independent prognostic factor. We investigated whether resistance to chemotherapy be represented by gene-expression profiles, and which genes are associated with resistance. DESIGN AND METHODS: cDNA microarrays containing approximately 41,000 features were used to compare the gene-expression profile of AML blasts between 33 patients with good or poor response to induction chemotherapy. Data generated by cDNA-arrays were confirmed by quantitative reverse transcription polymerase chain reaction. RESULTS: Using significance analysis of microarrays, we identified a characteristic gene-expression profile which distinguished AML samples from patients with good or poor responses. In hierarchical clustering analysis poor responders clustered together with normal CD34+ cells. Moreover, 13/40 (32.5%) genes highly expressed in poor responders are also overexpressed in hematopoietic stem/progenitor cells. Prediction analysis using 10-fold cross validation revealed an 80% overall accuracy. Using the treatment-response signature to predict the outcome in an independent test set of 104 AML patients, samples were separated into two subgroups with significantly inferior response rate (43.5% vs. 66.7%, p=0.04), significantly shorter event-free and overall survival (p=0.01 and p=0.03, respectively) in the poor-response compared to in the good-response signature group. In multivariate analysis, the treatment response signature was an independent prognostic factor (hazard ratio, 2.1, 95% confidence interval 1.2 to 3.6, p=0.006). INTERPRETATION AND CONCLUSIONS: Resistance to chemotherapy in AML can be identified by gene-expression profiling before treatment and seems to be mediated by a transcriptional program active in hematopoietic stem/progenitor cells. PMID- 16266897 TI - Population-based age-specific incidences of cytogenetic subgroups of acute myeloid leukemia. AB - BACKGROUND AND OBJECTIVES: It is well known that the different cytogenetic subgroups of acute myeloid leukemia (AML) show different age-specific frequencies. For example, balanced translocations tend to be found in younger patients while complex aberrant karyotypes are usually found in elderly patients with AML. However, detailed data on the population-based age-dependent incidences of distinct cytogenetic subtypes as well as of molecular mutations are lacking. DESIGN AND METHODS: We evaluated the population-based age-specific incidences of different cytogenetic subgroups in 2555 patients with AML between 21 and 70 years of age. We also investigated the association of specific molecular markers (FLT3 M, FLT3- TKD, MLL-PTD, NRAS, CEPBA, KITD816). RESULTS: The incidence of balanced translocations was rather constant over lifetime. In contrast, the incidence of unbalanced aberrations and especially complex aberrant karyotypes increased sharply with age. There were also different age-specific incidences of some recurrent molecular mutations. INTERPRETATION AND CONCLUSIONS: These results are suggestive of different mechanisms in the pathogenesis of AML. PMID- 16266896 TI - Quantification of AML1-ETO fusion transcript as a prognostic indicator in acute myeloid leukemia. AB - BACKGROUND AND OBJECTIVES: In spite of the high complete remission rate that chemotherapy achieves in acute myeloid leukemia with AML1-ETO gene rearrangement, relapse is a major cause of treatment failure in this condition. We aimed to determine a predictor of relapse with the real-time quantitative reverse transcription polymerase chain reaction (RQ-PCR) of AML1-ETO chimeric mRNA. DESIGN AND METHODS: We serially monitored AML1-ETO fusion transcripts using RQ PCR in 113 bone marrow or peripheral blood samples from 21 patients with AML1-ETO positive acute myeloid leukemia and analyzed the prognostic relevance of the results. RESULTS: Higher transcript levels at diagnosis were associated with a higher probability of relapse (p=0.038 in all patients and p=0.001 in adult patients). A decrease of less than 3-log at the time of achieving complete remission was also associated with a higher risk of relapse (p=0.035 in all patients and p=0.011 in adult patients). RQ-PCR detected the reappearance of AML1 ETO fusion transcripts in both peripheral blood and bone marrow during apparent complete remission. Detection of the transcripts preceded hematologic relapse by one to three months. The transcript levels in peripheral blood correlated with those in bone marrow at the same time point. INTERPRETATION AND CONCLUSIONS: Our findings indicate that regular monitoring of AML1-ETO chimeric transcript levels by RQ-PCR on bone marrow or peripheral blood samples could be extremely useful for the selection of high-risk patients and be an early predictor of relapse. PMID- 16266898 TI - The association of a distinctive allele of NAD(P)H:quinone oxidoreductase with pediatric acute lymphoblastic leukemias with MLL fusion genes in Japan. AB - BACKGROUND AND OBJECTIVES: The enzyme NAD(P)H:quinone oxidoreductase (NQO1) detoxifies chemicals with quinone rings including benzene metabolites and flavonoids. Previous studies in Caucasian populations have provided evidence that a loss of function allele at nt 609 (C609T, Pro187Ser) is associated with increased risk of infant acute lymphoblastic leukemia (ALL) with MLL-AF4 fusion genes. DESIGN AND METHODS: We genotyped 103 infants (<18 months) with ALL or acute myeloid leukemia (AML) in Japan and 185 controls for the frequency of allelic variation at nt 609 and 465 in NQO1 using standardized polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) methodology. RESULTS: The C609T polymorphism is very common in Japan but we found no link with altered risk for infant ALL. However, a variant of another allele at nt 465 (C465T, Arg139Trp), also associated with diminished enzyme activity, was strongly associated (OR 6.36; CI 1.84-21.90; p=0.002) with infant ALL, especially in t(4;11)(q21;q23), MLL-AF4. No association was found between this allele and risk of infant AML with MLL gene fusions or infant ALL without MLL gene fusions. The same C465T allele has been linked recently, in an Oriental population, to sensitivity to benzene hematotoxicity. INTERPRETATION AND CONCLUSIONS: These data endorse the notion that infant ALL with MLL fusion genes have a unique etiology possibly involving transplacental exposure to chemicals. PMID- 16266899 TI - Four-color flow cytometry bypasses limitations of IG/TCR polymerase chain reaction for minimal residual disease detection in certain subsets of children with acute lymphoblastic leukemia. AB - BACKGROUND AND OBJECTIVES: Competitive immunoglobulin/T-cell receptor polymerase chain reaction (PCR) analysis with fluorescent detection is a rapid, cheap and reproducible method for quantifying minimal residual disease (MRD), which is well adapted to the recognition of high-risk childhood acute lymphoblastic leukemia (ALL). We aimed at defining whether flow cytometry (FC) techniques can bypass limitations of PCR for MRD determination. DESIGN AND METHODS: We analyzed 140 remission samples from 91 patients using both competitive PCR amplification of antigen-receptor genes and four-color FC identification of leukemia immunophenotype. These methods were chosen with the aim of detecting at least 0.1% blasts. RESULTS: MRD was measured using both PCR and FC methods in 123 samples and the two methods provided concordant results in 119 of them (97%). Moreover, three out of the four discordant results appeared minor since MRD was detectable by both methods, but at different levels. In 12 of 13 samples from nine patients, mainly infants with early CD10- and/or t(4;11) B-cell ALL and children with immature T-cell ALL, MRD could be determined using FC whereas PCR failed. Conversely, FC methods were unfeasible due to inappropriate leukemia immunophenotype in three additional children (including two with T-cell ALL) for whom PCR successfully provided MRD results. INTERPRETATION AND CONCLUSIONS: The MRD results provided by FC techniques were highly concordant with those of competitive PCR. Moreover, the applicability of FC appeared higher in certain ALL subsets, although the appropriateness of this technique in terms of outcome prediction remains to be demonstrated. PMID- 16266900 TI - Resolving T-cell receptor clonality in two and genotype in four multiplex polymerase chain reactions. AB - BACKGROUND AND OBJECTIVES: The diagnosis of T-cell neoplasia requires the use of immunohistochemistry on tumor sections or molecular genetic analysis of T-cell receptor (TCR) clonality. Multiplex polymerase chain reactions (PCR) offer a sensitive and expeditious approach to determining clonality early in the diagnostic work-up. We determined the sensitivity and specificity of four multiple PCR for genotyping lymphoid neoplasms at the TCR loci gamma (TCRG), delta (TCRD) and beta (TCRB, including complete [Vbeta-Jbeta] and incomplete [Dbeta-Jbeta] rearrangements). DESIGN AND METHODS: Template DNA was derived from frozen or formalin-fixed tissue and from imprints of aspirates or cut tissue surface on a FTA MicroCard. Each multiplex PCR was performed for 36 cycles in a single tube with multiple previously reported fluorescently labeled primers (TCRG and TCRD) or novel homologous primers (TCRB) and analyzed on electropherograms (Genescan), applying stringent criteria for interpreting clonal peaks. Two hundred and eleven clinically and immunohistochemically well-characterized benign and malignant non-T-cell lymphoid proliferations, including 138 B-cell lymphomas, were analyzed to determine specificity. The results were compared with those of 28 peripheral and immature T-cell neoplasms and two NK/T-cell lymphomas to determine sensitivity and compute predictive values. RESULTS: In all T-cell tumors, one or more TCR loci showed clonal rearrangement, which was not evident in two NK/T-cell lymphomas. TCRG was the single most informative locus (clonal rearrangement in 89%), followed by TCRB (79%) and TCRD (39%). Multiplex PCR targeting of TCRG and TCRD together resolved clonality in all T-cell neoplasms, whereas the TCRB locus was clonal in two of three cases with polyclonal TCRG. Unexpectedly, in B-cell lymphomas single clonal incomplete TCRB (Dbeta-Jbeta) peaks were 20 times more likely to occur than clonal TCRG. INTERPRETATION AND CONCLUSIONS: Clonality can be accurately determined in nodal T-cell lymphoma with two single-tube multiplex PCR targeting TCRG and TCRD. TCRB analysis should be considered in equivocal cases in which a polyclonal background may obscure clonal TCRD, but clonal incomplete TCRB rearrangement alone is insufficient for presuming T-cell lineage. In the absence of objective evidence of B-cell neoplasia, multiplex PCR of T-cell receptor genes may be used early in the diagnostic work-up, including for fine needle aspirates. PMID- 16266901 TI - Comparison of ZAP-70/Syk mRNA levels with immunoglobulin heavy-chain gene mutation status and disease progression in chronic lymphocytic leukemia. AB - BACKGROUND AND OBJECTIVES: The protein tyrosine kinase ZAP-70 has recently emerged as a major prognostic indicator in chronic lymphocytic leukemia (CLL). ZAP-70 is structurally and functionally homologous to Syk, a key mediator of B cell receptor signaling. We therefore evaluated ZAP-70 expression in CLL B cells using Syk as an intracellular standard. DESIGN AND METHODS: The relative amounts of ZAP-70 and Syk were determined in purified B cells from 92 CLL patients using a novel reverse transcriptase/polymerase chain reaction (RT-PCR) procedure that co-amplifies both transcripts with equal efficiency. The ZAP-70/Syk mRNA ratio was correlated with VH gene mutation status, median treatment-free survival and FACS analysis of ZAP-70 expression. RESULTS: ZAP-70 was expressed in the majority of cases with unmutated VH genes (88%), but also at lower levels in a substantial fraction of cases with mutated VH genes (44%). High levels of ZAP-70, defined as ZAP-70/Syk mRNA ratios above 0.25, were observed mainly in cases with unmutated VH genes and correlated with short treatment-free survival. In contrast, no difference was observed in the median treatment-free survival between patients with low ZAP-70/Syk ratios (0.05-0.25) and patients with no or negligible ZAP-70 expression (ZAP-70/Syk<0.05). In 73 cases ZAP-70 expression was investigated by RT/PCR and FACS analysis; concordance with VH gene mutation status was 86% and 71%, respectively. INTERPRETATION AND CONCLUSIONS: ZAP-70 is frequently expressed in CLL B cells, but only high levels correlate with unmutated VH gene status and progressive disease. Expression of ZAP-70 can be accurately assessed by analysis of the ZAP-70/Syk mRNA ratio, thus providing an alternative to FACS analysis. PMID- 16266903 TI - Hemostatic balance on the surface of leukemic cells: the role of tissue factor and urokinase plasminogen activator receptor. AB - BACKGROUND AND OBJECTIVES: The frequency of thrombotic complications is increased in patients with acute leukemia. Since coagulation processes take place on cell membranes, we hypothesized that expression of coagulation proteins on blast membrane could determine the hemostatic balance on the surface of leukemic cells and may correlate with thrombotic manifestations. DESIGN AND METHODS: Fifty-one consecutive patients with newly diagnosed acute leukemia were enrolled over an 11 month period. Twenty-five of the patients had acute myeloid leukemia (AML)-M(0 2), 11 had AML-M3, 6 had AML-M(4-5), and 9 acute lymphocytic leukemia (ALL). Peripheral blood and bone marrow were analyzed by flow-cytometry for tissue factor, protease-activated receptor 1, tissue factor pathway inhibitor, urokinase plasminogen activator receptor, and thrombomodulin. RESULTS: Regardless of the leukemia subtype, tissue factor was predominantly present on leukemic blast surfaces as compared to protease-activated receptor 1, tissue factor pathway inhibitor, urokinase plasminogen activator receptor and thrombomodulin and it was significantly elevated (mean 63+/-6%) in AML-M3 and AML-M(4-5) as compared to AML M0(-2) and ALL (mean 37+/-4%, p<0.001). Likewise, urokinase plasminogen activator receptor expression was greater in AML-M(4-5) (49+/-11%) than in in AML- M(0-2), M3 and, ALL (mean 17+/-3%, p<0.001). Thrombotic manifestations were present in 13 out of 51 (26%) patients. The tissue factor to urokinase plasminogen activator receptor ratio was higher in patients with a thrombotic event than in patients without thrombotic events (16+/-4 vs. 6+/-2, p=0.042). INTERPRETATION AND CONCLUSIONS: This study demonstrates that tissue factor predominates on leukemic blast surface, particularly in M3 and M4-5 subtypes, while urokinase plasminogen activator receptor is increased on M4-5 blasts. The hemostatic balance on the blast surface may contribute to thrombotic manifestations in leukemic patients. PMID- 16266902 TI - Molecular and biological characterization of three novel interleukin-6-dependent human myeloma cell lines. AB - BACKGROUND AND OBJECTIVES: Established human myeloma cell lines (HMCL) have significantly contributed to the investigation of the biological aspects of multiple myeloma. Our study reports the molecular and biological characterization of three novel interleukin-6 (IL-6)-dependent HMCL (CMA-01, CMA-02, CMA-03) established from the malignant plasma cells of myeloma patients with extramedullary disease. DESIGN AND METHODS: The immunophenotype, cell growth characteristics, IL-6 pathway, chromosomal alterations and gene expression profiles of the three HMCL were investigated. RESULTS: The plasma cell origin of the three Epstein-Barr virus-negative HMCL was confirmed by immunophenotypic analysis. Cytogenetic and fluorescence in situ hybridization analyses revealed the presence of complex karyotypes with many numerical and structural chromosomal abnormalities. All three HMCL are positive for the t(8;14); CMA-01 and CMA-02 showed t(11;14) and t(14;16) translocations, respectively. The three HMCL grow slowly at a relatively low saturation density and depend on exogenous IL-6 for their survival and proliferation. The comparison of the gene expression profiles of the three HMCL versus those of the purified tumor plasma cells from which the cell lines were derived identified a set of differentially expressed genes mainly involved in the cell proliferation pathway. INTERPRETATION AND CONCLUSIONS: Extensively characterized large HMCL panels that reflect the heterogeneity of the disease may improve our understanding of the pathogenetic events and clinical progression of multiple myeloma. PMID- 16266904 TI - Haptoglobin phenotype 2-2 as a potentially new risk factor for spontaneous venous thromboembolism. AB - BACKGROUND AND OBJECTIVES: Haptoglobin (Hp) is a plasma protein that binds free hemoglobin and thus prevents catalysis of reactive oxygen species by the Fenton reaction. A genetic polymorphism has been described that leads to the generation of two distinct alleles, Hp1 and Hp2, which define three major haptoglobin phenotypes, denoted Hp1-1, Hp2-1 and Hp2-2. Hp2-2 has been reported to be associated with the risk of atherosclerosis and coronary heart disease. In our study we investigated the association of haptoglobin genotype and phenotype with the risk of spontaneous venous thromboembolism (VTE). DESIGN AND METHODS: One hundred and twenty-eight patients with a history of spontaneous deep vein thrombosis (70 women, 58 men), 105 with spontaneous symptomatic pulmonary embolism (58 women, 47 men) and 122 healthy controls (60 women, 62 men) were enrolled. Haptoglobin levels were measured immunonephelometrically and phenotypes were detected by polyacrylamide gel electrophoresis and subsequent immunoblotting. RESULTS: The Hp2-2 phenotype was significantly more prevalent in patients (42%) than in controls (30%) and significantly increased the risk for VTE in univariable (odds ratio=1.6, 95% confidence interval [1.0-2.6], p=0.04) and multivariable analyses (odds ratio=1.9 [1.0-3.4], p=0.04). Hp2-2 (n=134) was associated with significantly lower haptoglobin levels (median=89.7 mg/dL) than Hp2-1 (n=170, median = 123.5 mg/dL, p<0.001) or Hp1-1 (n=51, median=142.8 mg/dL, p<0.001). INTERPRETATION AND CONCLUSIONS: Our study gives the first evidence that Hp2-2 represents a risk factor for spontaneous VTE, presumably through a pathophysiological mechanism similar to that in arterial disease. PMID- 16266905 TI - The impact of donor factors on primary non-engraftment in recipients of reduced intensity conditioned transplants from unrelated donors. AB - BACKGROUND AND OBJECTIVES: Primary graft failure is a serious complication following hematopoietic cell transplants, particularly when using unrelated donors. We analyzed factors affecting primary graft failure in recipients of hematopoietic cell transplants from unrelated donors, which were performed using reduced intensity conditioning. DESIGN AND METHODS: This was a retrospective analysis of 144 patients whose transplants took place between March 1998 and October 2004. The data were analyzed in January 2005. RESULTS: The median age of the patients was 51 years. The diagnoses were varied. Conditioning regimens were fludarabine, melphalan, campath (n=80), fludarabine, busulphan, campath (n=38), fludarabine, BEAM, campath (n=9) and other (n=17). The donor was 10/10 allele matched in 95/144 (66%) cases; 94 donated bone marrow and 50 peripheral blood stem cells. The 3-year probability of overall survival was 43%. The median follow up was 724 days (range: 91-1651 days). Of evaluable patients, 7/140 (5%) failed to achieve myeloid engraftment. Primary graft failure was significantly associated with the use of a mismatched donor (6/47,13% versus 1/93, 1%, p=0.006), as well as: bone marrow as the source of stem cells (p=0.046), chronic myeloid leukemia compared to other diagnoses (p=0.022), and a female rather than a male donor (p=0.019). In multivariate analysis chronic myeloid leukemia, HLA mismatched and/or female donors remained significantly associated with primary graft failure. Single HLA mismatches were tolerated, however in multiply mismatched grafts, overall survival was worse (p=0.005); transplanted-related mortality (p=0.005) and chronic graft-versus-host disease (p=0.025) were increased. INTERPRETATION AND CONCLUSIONS: These data have implications for the choice of donor and stem cell source in transplants performed using reduced intensity conditioning regimens, suggesting that the use of bone marrow, female donors and HLA-mismatched grafts increase the risk of primary graft failure, and should be avoided in certain situations. PMID- 16266907 TI - Effects of the protein tyrosine kinase inhibitor, SU5614, on leukemic and normal stem cells. AB - FLT3 activating mutations are the most frequent single genetic abnormality in patients with acute myeloid leukemia. Thus targeting the FLT3 activated kinase is a promising treatment approach. We wanted to test whether the protein tyrosine kinase inhibitor SU5614 selectively eliminates leukemic stem cells while sparing their normal counterparts. PMID- 16266906 TI - Inhibition of coagulation by macromolecular complexes. AB - The role of vertebrate blood coagulation is to rapidly prevent the loss of body fluids following vascular injury without compromising blood flow through either the uninjured or damaged vessels. To achieve this the coagulation network is initiated and regulated by a complex network of interactions that are under the control of both positive and negative feedback loops that result in controlled fibrin deposition and platelet activation only at the site of injury. Anticoagulant molecules play key roles in preventing inappropriate initiation of coagulation as well as down-regulating thrombin generation at the site of injury. Tissue factor pathway inhibitor (TFPI) inhibits the initiation complex, antithrombin (AT) inhibits the active serine proteases directly, whereas the activated protein C pathway inhibits coagulation by inactivating the cofactors V and VIII. In this review the structure and function of these anticoagulant molecules and their inhibitory complexes is discussed. PMID- 16266908 TI - Rituximab in patients with mucosal-associated lymphoid tissue-type lymphoma of the ocular adnexa. AB - Eight patients with ocular adnexal mucosal-associated lymphpid tissue (MALT) lymphoma were treated with rituximab, at diagnosis (n=5) or relapse (n=3). All untreated patients achieved lymphoma regression, while relapsing patients had no benefit. Four responding patients experienced early relapse. The median time to progression was 5 months. The efficacy of rituximab in ocular adnexal lymphoma is lower than that reported for gastric MALT lymphomas. PMID- 16266909 TI - High-dose therapy with autologous stem cell transplantation in first response in mantle cell lymphoma. AB - We retrospectively investigated the outcome of 30 newly diagnosed patients with mantle cell lymphoma treated with high-dose therapy and autologous stem cell transplantation in first response. With a median follow-up of 55 months, the 5 year overall-survival is 62%, the 5-year progression-free-survival is 40% and no secondary malignancy has occurred. PMID- 16266910 TI - Development of functional Haemophilus influenzae type b antibodies after vaccination. AB - Sixteen autologous stem cell transplant recipients received three vaccinations with conjugated haemophilus influenzae type b vaccine. Quantitative and qualitative aspects of the antibody response were studied. The vaccination schedule resulted in high antibody response rates and functional maturation of antibodies, as measured by antibody avidity and phagocytosis-inducing capacity. PMID- 16266911 TI - A 4 base pair TGAT insertion at codon 116 of the beta globin gene causes beta0 thalassemia. AB - A new beta(0) thalassemia allele caused by a TGAT insert in codon 116 of exon III was detected in a patient compound heterozygous for beta(0) thalassemia / Hb D Los Angeles and his father. The mutation unexpectedly causes a classical thalassemic phenotype. The compound heterozygosity leads to mild microcytic anemia and no further clinical signs. PMID- 16266912 TI - Reduction of glycosylated hemoglobin with stable insulin levels in a diabetic patient with chronic myeloid leukemia responsive to imatinib. PMID- 16266913 TI - Mucormycoses in patients with hematologic malignancies: an emerging fungal infection. AB - BACKGROUND AND OBJECTIVES: Mucormycoses are seen with an increasing incidence in immunocompromised patients. Most common presentations are rhinocerebral and pulmonary. We here report the experience of a single center with mucormycoses in patients with hematologic malignancies. RESULTS: Mucormycoses were diagnosed in six patients, (median age of 52 years; range, 26-74) treated between 2001-2004. Diagnoses included acute myeloid leukemia (AML) (n=3), acute lymphoblastic leukemia (n=1), chronic lymphocytic leukemia (n=1) and multiple myeloma (n=1). Mucormycosis was diagnosed in the neutropenic state following allogeneic hematopoietic cell transplantation (n=3) or intense chemotherapy (n=3). Sites of infections were rhinocerebral, facial and pulmonary involvement in one patient each and disseminated mucormycosis in three patients. The diagnosis was established by computed tomography followed by surgical interventions and histological diagnosis in 4 patients and post-mortem in two patients. Species identified were Rhizopus (n=3), Rhizomucor (n=2) and Absidia (n=1). Treatment responses were best if surgical resection was followed by aggressive antifungal chemotherapy. Five of six 6 patients died, all of complications of mucormycosis or their underlying disease. Only one patient with facial mucormycosis is still alive. CONCLUSIONS: This experience demonstrates that patient with mucormycoses have a high mortality rate and early recognition followed by aggressive surgical debridement, high dose antifungal therapy and attempts to correct the underlying immunocompromised state are crucial in the treatment of this fatal infection. PMID- 16266914 TI - Rituximab-induced acute thrombocytopenia: a report of two cases. AB - Rituximab use in B-cell malignancies has been widely favored by the acceptable toxicity profile of this drug. Episodes of rituximab-induced neutropenia have been reported in some patients, but severe acute thrombocytopenia is very unusual. Here, we report transient severe acute thrombocytopenia after rituximab infusion in two patients with, one hairy cell leukemia the other with mantle cell lymphomay. Interestingly, in both cases, thrombocytopenia was reversible in a few days without further therapeutic intervention. The mechanism of this side effect remains unclear. Previous reports suggested the presence of CD20 antigen on the platelets themselves or that soluble CD20 antigen in the circulation may cause an antigen-antibody reaction and immune-mediated cell lysis. It is noteworthy that the two cases reported here as well as the two previously published cases share massive bone marrow involvement by neoplastic B lymphocytes. PMID- 16266915 TI - Optic nerve relapse in a child with common acute lymphoblastic leukemia, treated with systemic anti-CD-20 (rituximab). AB - We describe a two- year old boy who was diagnosed with pre-B acute lymphoblastic leukemia (ALL). He developed a central nervous system (CNS) relapse with optic nerve involvement. Initially he was treated according to the ALL relapse protocol, including CNS radiotherapy. Despite an initial complete reponse, relapse occurred within six weeks of treatment. The leukemic blast cells were CD 20 positive and he was treated with systemic anti CD-20 therapy (rituximab) with no CNS recurrence over a six-month period. He died due to a CD-20 negative bone marrow relapse. This case illustrates a potential role for rituximab in pediatric CD-20 positive malignancies. PMID- 16266916 TI - Clinical significance of development of Philadelphia-chromosome negative clones in patients with chronic myeloid leukemia treated with imatinib mesylate. PMID- 16266917 TI - Complete remission of hairy cell leukemia variant (HCL-v) complicated by red cell aplasia post treatment with rituximab. AB - Hairy cell leukemia variant (HCL-v) is a rare form of a chronic B-cell lymphoproliferative disorder. Unlike typical hairy cell leukemia (HCL) where the complete response (CR) rate to 2-chlorodeoxyadenosine and 2'-deoxycoformycin can approach about 90%, in HCL-v CR is rare and partial response (PR) occurs in approximately 50% with these agents. Rituximab treatment in relapsed or refractory HCL results in a CR of 13% to 53%, but its use in HCL-v has not been reported in the literature to our knowledge. We describe a patient with HCL-v, whose course was previously complicated by pure red cell aplasia who achieved CR after treatment with rituximab. We also briefly review outcomes of treatments used in HCL-v reported in the current literature. PMID- 16266918 TI - Post-transplantation lymphoproliferative disorder of recipient origin in a boy with acute T-cell leukemia with detection of B-cell clonality 3 months before stem cell transplantation. AB - Post-transplantation lymphoproliferative disorder is an infrequent complication after hematopoietic stem cell transplantation. It is hypothesized that lack of T cell surveillance following transplantation permits reactivation of latent EBV leading to polyclonal B-cell expansion and finally outgrowth of a predominant clone. Most cases are of donor origin. Here, we describe an 8-year old boy with early onset post-transplantation lymphoproliferative disorder following matched unrelated stem cell transplantation for high-risk T-cell leukemia whose disease was unusual for two reasons. First, his B-cell clone was of host origin and, in contrast to the few PTLD of host origin described so far, not associated with autologous reconstitution. Secondly, using clonal analysis, we could retrospectively show that the B-cell clone emerged during consolidation chemotherapy for T-cell leukemia, 3 months before stem cell transplantation. PMID- 16266919 TI - Cerebellar degeneration and folate deficiency due to cough mixture abuse. PMID- 16266920 TI - Kasabach-Merritt syndrome associated with giant liver hemangioma: the effect of combined therapy with danaparoid sodium and tranexamic acid. AB - n patients with Kasabach-Merritt syndrome (KMS), local activation of coagulation commonly results in disseminated intravascular coagulation (DIC). Progress of DIC is associated with 30-40% mortality as a result of uncontrollable hemorrhage. A 39-year-old woman with an enlarging giant liver hemangioma was diagnosed as having KMS with DIC. To control the hemorrhagic diathesis, we commenced combination therapy for DIC with danaparoid (1,250 Ux2/day, intravenously (IV)) and tranexamic acid (0.5 g x 3/day, peros (PO). Rapid improvement of the bleeding tendency and coagulopathy occurred in response to this treatment - that is, DIC was controlled without removing the giant hemangioma. The therapy did not restrict the behavior of the patient by continuous drip and angiography could be performed without bleeding. Such therapy may be beneficial in chronic DIC with activation of fibrinolysis. PMID- 16266921 TI - Venous thromboembolism in a young woman with combined homozygosity for factor V Leiden and prothrombin G20210A mutations. PMID- 16266922 TI - Membranous glomerulopathy in children given allogeneic hematopoietic stem cell transplantation. AB - Graft-versus-host disease (GVHD) is a common complication of allogeneic hematopoietic stem cell transplantation (HSCT), but membranous glomerulopathy (MG) has rarely been described as a manifestation of chronic GVHD. We report two cases of MG in children who underwent allogeneic HSCT. The clinical findings were characterized by edema of the lower extremities and nephrotic proteinuria in one case and hypertension, hematuria and edema with non-nephrotic proteinuria in the other one. Renal biopsy was consistent with MG and appropriate immunosuppressive therapy was prescribed. Both patients achieved complete remission and are alive without renal disease 4 and 2 years after the diagnosis of MG. The normal levels of albumin and non-nephrotic proteinuria in one of the two cases raise the question of whether the real incidence of MG after HSCT is underestimated. Therefore, we strongly suggest regular urine analysis during the follow-up of children undergoing HSCT in order to diagnose MG early. PMID- 16266923 TI - Paravertebral extramedullary hematopoiesis due to pyruvate kinase deficiency. PMID- 16266924 TI - Successful treatment of pure red cell aplasia with repeated, low doses of rituximab in two patients after ABO-incompatible allogeneic haematopoietic stem cell transplantation for acute myeloid leukaemia. AB - We describe two patients with acute myeloid leukemia successfully treated with anti-CD20 antibody for pure red cell aplasia (PRCA) following ABO-mismatched allogeneic hematopoietic stem cell transplantation (HSCT). PRCA following HSCT is associated with major ABO incompatibility between donor and recipient and is due to an inhibition of donor erythroid precursors by residual host isoagglutinins. The first patient developed PRCA resistant to several treatment options including donor-derived leukocyte infusions (DLI), high-dose erythropoietin (EPO), and rapid tapering of cyclosporin A (CsA). This patient also received anti-viral therapy as CMV and parvovirus B19 infections were regarded as additional causes of PRCA. Due to a loss of donor chimerism, he underwent second HSCT, but PRCA still persisted. He showed no evidence of graft-versus-host disease (GVHD). Finally he was administered anti-CD20 antibody (rituximab) at a dose of 150/m2 and PRCA resolved in a short period of time. The case was complicated by life threatening pulmonary aspergillosis with septic shock, successfully treated with anti-fungal therapy. The second case concerns a patient, who revealed PRCA after major ABO-incompatible HSCT from his brother. Considering our experience with the previously described patient, he proceeded to rituximab at a dose of 150/m2 as first line treatment. We observed rapid recovery from PRCA without any side effects. We conclude that rituximab seems to be a promising therapeutic option in patients with PRCA after ABO-mismatched HSCT, in whom conventional treatment fails. PMID- 16266925 TI - Are thrombotic complications in patients with von Willebrand's disease expression of a multifactorial disease? PMID- 16266926 TI - Human herpesvirus 8 and Epstein-Barr virus coinfection in localized Castleman disease during pregnancy. AB - Castleman's disease is a rare disorder characterized by two distinct entities with similar histology but different time course and therapeutic response. Multicentric plasma cell variant is highly associated with infection by human herpesvirus 8 (HHV-8), but the pathogenesis of the hyaline vascular variant is currently unknown. We report a pregnant patient who develops a localized axillary hyaline-type Castleman's disease in which HHV-8 DNA sequences were detected in the lymph node lesions by nested PCR. In addition, the PCR multiplex also showed positivity for EBV. Immunohistochemical studies confirmed the presence of both viruses. Our results provide the first evidence of the presence of HHV-8 and EBV sequences in localized Castleman's disease, suggesting a possible role of the association of these herpes virus in the pathogenesis of this type of disorder. This case highlights that searching for HHV-8 and EBV sequences in cases of localized Castleman's disease is strongly advised. PMID- 16266927 TI - Mobilization of autologous hematopoietic progenitors and subsequent transplantation is a safe and feasible procedure in chronic phase chronic myelogenous leukemia patients with cytogenetic resistance to imatinib. PMID- 16266928 TI - Remission of severe antiphospholipid syndrome associated with non-Hodgkin's B cell lymphoma after combined treatment with rituximab and chemotherapy. AB - The association of lymphoid neoplasms and antiphospolipid antibodies (APA), with or without thromboembolic complications, has been reported in several cases. We describe one case of B-cell non-Hodgkinis lymphoma (NHL) in which the combination of rituximab with standard chemotherapy led to the complete remission of a severe hypercoagulable state associated with APA. PMID- 16266929 TI - Premature labor and leukoerythroblastosis in a newborn with parvovirus B19 infection. AB - Leukoerythroblastosis is a rarely observed disease characterized by the presence of leukocytosis, erythroid and myeloid blast cells in peripheral blood. To our knowledge, it had not been diagnosed in a premature newborn before the case we report have.A female baby weighing 1164 grams, who was born prematurely at the 29th week of gestation by Cesarean section was referred to our newborn intensive care unit due to prematurity and respiratory distress with no prenatal pathological findings. Physical examination revealed tachypnea and hepatosplenomegaly. Routine laboratory measurements showed significant leukocytosis (85,000/mm3) and anemia (Hb: 9.6 g/dL and Hct: 27.6%). The platelet count was normal. The peripheral blood smear suggested leukoerythroblastosis with the presence of nucleated erythrocytes, monocytosis, and 4% blasts. Bone marrow cytogenetic examination was normal. Parvovirus B19 Ig G and M serology were detected to be positive. The etiological factors observed in leukoerythroblastosis occurring during neonatal and early childhood period are congenital-postnatal viral infections, juvenile myelomonocytic leukemia and osteopetrosis. To our knowledge, no case of leukoerythroblastosis in such an early phase has been reported in the in literature. As a result, premature delivery and leukoerythroblastosis were thought to have developed secondary to intrauterine parvovirus B19 infection. Leukoerythroblastosis is a rarely observed disease characterized by the presence of leukocytosis, erythroid and myeloid blast cells in peripheral blood. It is reported that it can be observed following hematologic malignancies especially juvenile myelomonocytic leukemia, acute infections, hemolytic anemia, osteopetrosis, myelofibrosis, neuroblastoma and taking certain medicines. To our knowledge, it has not been diagnosed in a premature newborn before. Here we the case of a newborn who was referred to our intensive care unit due to being born prematurely at the 29th week of gestation and diagnosed with leukoerythroblastosis. PMID- 16266930 TI - Primary gastrointestinal lymphoma of the colon. PMID- 16266931 TI - Binding of alpha-1-acid glycoprotein to imatinib following increased dosage of drug. PMID- 16266933 TI - Brief comment on the case report by Fattori et al. 2005 (5) 589-590. PMID- 16266936 TI - Home-treatment of deep vein thrombosis in patients with cancer: quality of life impact of therapies. PMID- 16266934 TI - The role of serum erythropoietin levels in diagnosis and classification of erythrocytosis/polycythemia. PMID- 16266937 TI - Safety and economic impacts of photo radar program. AB - OBJECTIVE: Unsafe speed is one of the major traffic safety challenges facing motorized nations. In 2003, unsafe speed contributed to 31 percent of all fatal collisions, causing a loss of 13,380 lives in the United States alone. The economic impact of speeding is tremendous. According to NHTSA, the cost of unsafe speed related collisions to the American society exceeds 40 billion US dollars per year. In response, automated photo radar speed enforcement programs have been implemented in many countries. This study assesses the economic impacts of a large-scale photo radar program in British Columbia. The knowledge generated from this study could inform policy makers and project managers in making informed decisions with regard to this highly effective and efficient, yet very controversial program. METHODS: This study establishes speed and safety effects of photo radar programs by summarizing two physical impact investigations in British Columbia. It then conducts a cost-benefit analysis to assess the program's economic impacts. The cost-benefit analysis takes into account both societal and funding agency's perspectives. It includes a comprehensive account of major impacts. It uses willingness to pay principle to value human lives saved and injuries avoided. It incorporates an extended sensitivity analysis to quantify the robustness of base case conclusions. RESULTS: The study reveals an annual net benefit of approximately 114 million in year 2001 Canadian dollars to British Columbians. The study also finds a net annual saving of over 38 million Canadian dollars for the Insurance Corporation of British Columbia (ICBC) that funded the program. These results are robust under almost all alternative scenarios tested. The only circumstance under which the net benefit of the program turns negative is when the real safety effects were one standard deviation below the estimated values, which is possible but highly unlikely. CONCLUSION: Automated photo radar traffic safety enforcement can be an effective and efficient means to manage traffic speed, reduce collisions and injuries, and combat the huge resulting economic burden to society. The cost-effectiveness of the program takes on special meaning and urgency when considering the present and future government funding constraints. The application of the program, however, should be planned and implemented with caution. Every effort should be made to focus on and to promote the program on safety improvement grounds. The program can be easily terminated because of political considerations, if the public perceives it as a cash cow to enhance government revenue. PMID- 16266938 TI - Quirks of mass accident data bases. PMID- 16266939 TI - Multilevel modeling in traffic safety research: two empirical examples illustrating the consequences of ignoring hierarchies. AB - OBJECTIVES: This commentary illustrates the advantages of multilevel modeling compared to statistical techniques that ignore hierarchies, based on two empirical traffic safety examples. METHODS: The common concept shared by different definitions of multilevel modeling is identified and illustrated. Each definition defines multilevel modeling in its own way but they all refer to hierarchies. Conceptual issues inherently related to hierarchies are then pointed out and illustrated. RESULTS: Broadly speaking there are two important consequences of ignoring a hierarchical structure in the data. The first consequence, underestimation of standard errors, is illustrated with data from an observational study on seatbelt behavior. Two effects that were significant at the 5% level in a single-level model were no longer found to be significant in a two-level model. The single-level model is therefore bound to lead to erroneous conclusions regarding variables that could have an impact on seatbelt use and, ultimately, on increasing the level of traffic safety. The second consequence, related to contextual information, is illustrated with data from a roadside survey on drink driving. Of particular interest is the relationship between Traffic Count, an aggregated level 2 explanatory variable and Odds of drink driving, an individual level 1 dependent variable. CONCLUSIONS: Like every statistical technique, multilevel models should be used with caution and reservation. However, given certain limitations, multilevel modeling is very useful and valuable to traffic safety research. PMID- 16266940 TI - Driving under the influence of drugs in Sweden with zero concentration limits in blood for controlled substances. AB - OBJECTIVE: This article describes the background and implementation in Sweden of zero-concentration limits for controlled drugs in the blood of drivers. Eliminating the need to prove that a person's ability to drive safely was impaired by drugs has greatly simplified the prosecution case, which now rests primarily on the forensic toxicology report. Driving under the influence of a prescription drug listed as a controlled substance is exempt from the zero-limit law provided the medication was being used in accordance with a physician's direction and the person was not considered unfit to drive. METHODS: The prevalence of driving under the influence of drugs (DUID) in Sweden was evaluated from police reports with the main focus on the toxicological findings. A large case series of DUID suspects was compared before and after introducing zero concentration limits in blood for controlled substances on July 1, 1999. The spectrum of drugs used by typical offenders and the concentrations of various licit and illicit substances in blood were evaluated and compared. RESULTS: Immediately after the zero-limit law came into force, the number of cases of DUID submitted by the police for toxicological analysis increased sharply and is currently ten-fold higher than before the new legislation. Statistics show that about 85% of all blood samples sent for toxicological analysis have one or more banned substances present. Amphetamine is by far the leading drug of abuse in Sweden and was identified in about 50-60% of all DUID suspects either alone or together with other controlled substances. The next most frequently encountered illicit drug was tetrahydrocannabinol (THC), with positive findings in about 20 25% of cases. Various prescription drugs, mainly sedative-hypnotics like diazepam and flunitrazepam, were also highly prevalent and these occurred mostly together with illicit substances. Opiates, such as 6-acetyl morphine and morphine, the metabolites of heroin, were high on the list of substances identified. Most DUID suspects in Sweden were men (85%) who were poly-drug users combining illicit substances, like amphetamine and/or cannabis, with a prescription medication such as various benzodiazepines. CONCLUSIONS: Sweden's zero-concentration limit has done nothing to reduce DUID or deter the typical offender because recidivism is high in this population of individuals (40-50%). Indeed, many traffic delinquents in Sweden are criminal elements in society with previous convictions for drunk and/or drugged driving as well as other offenses. The spectrum of drugs identified in blood samples from DUID suspects has not changed much since the zero-limit law was introduced. PMID- 16266942 TI - Influential factors in freeway crash response and clearance times by emergency management services in peak periods. AB - OBJECTIVE: Various factors influence the time performance of emergency management personnel when a freeway traffic crash occurs. The proper identification and prioritization of factors that contribute to emergency management services' response times and clearance times result in better usage of taxpayer resources. METHOD: Use of a proportional hazard-based Cox-regression model analyzed statewide, peak-period, traffic crash data from 1999 Ohio logs. These data included time performance measures of emergency management services. RESULTS: Traffic crash severity had the most effect on response times. Those crashes involving injuries or fatalities had up to 20% less emergency management service response times than "property damage only" crashes. Environmental factors such as weather or roadway conditions had minimal effect on response times to traffic crashes. Day of week, urban or rural area, off or opposing-lane crash location, number of vehicles involved, heavy vehicle involvement, and response time significantly affected clearance time and the resulting total time during peak periods. CONCLUSIONS: By assessing resources currently dedicated to insignificant factors, emergency management services can further improve response times to those casualties that crucially need emergency services. By accurately identifying and deciphering traffic crash severity from initial field reports, services can further improve. Moreover, improvements in crash severity prediction reduce "false alarms" for emergency services. The improvements reduce the probability of a very short response time for a property damage only crash in which initial reports implied a very severe injury. By focusing on factors that significantly reduce traffic crash clearance times on freeways in peak periods, more reductions in average delay experienced by freeway users, in fuel consumption, and in motor vehicle emissions can occur. PMID- 16266941 TI - The effect of a campus-community environmental alcohol prevention initiative on student drinking and driving: results from the "a matter of degree" program evaluation. AB - OBJECTIVE: Motor vehicle crashes involving alcohol are a major contributor to morbidity and mortality among college students in the United States. This study evaluates the effect on drinking and driving outcomes of the "A Matter of Degree" program, a campus-community coalition initiative to reduce college binge drinking. METHODS: We used a quasi-experimental longitudinal study design that compared student responses at 10 colleges participating in the program and students attending 32 similar colleges that did not participate in the program. We also divided the program sites into two groups of five according to their level of program implementation and compared each with the non-program colleges. We examined driving after any alcohol consumption and driving after five or more drinks among drinkers who drove one or more times a week per month and riding with a high or drunk driver among all students at these colleges beginning in 1997 through 2001. Outcomes were based on data collected from repeated cross sectional surveys using the Harvard School of Public Health College Alcohol Study. Analyses were conducted using MLwiN multilevel statistical software. RESULTS: We found significant reductions in driving after drinking, driving after five or more drinks and riding with a high or drunk driver at the program colleges relative to the comparison colleges. Further analyses indicated that these reductions among the AMOD program colleges occurred at the sites with high program implementation relative to comparison sites, while no statistically significant change was noted at the program sites with low implementation. The program effect on the two drinking and driving outcomes appeared to be mediated by frequent binge drinking, while significant decline in the riding with an intoxicated driver outcome was not mediated by the individual's drinking. CONCLUSIONS: Campus-community based environmental alcohol prevention is a promising approach for reducing alcohol-impaired motor vehicle crashes among this population. PMID- 16266943 TI - Frontal impact dummy kinematics in oblique frontal collisions: evaluation against post mortem human subject test data. AB - OBJECTIVE: Today, a predominant percentage of vehicles involved in car crashes are exposed to oblique or frontal offset collisions. The aim of this study is to evaluate the 50th percentile male Hybrid III, THOR 99, and THOR Alpha dummies by comparing them with the corresponding kinematics of post mortem human subjects (PMHS) in this type of collision. METHODS: The PMHS data include results from oblique frontal collision tests. They include sled tests with near-side and far side belt geometries at 15 degrees , 30 degrees , and 45 degrees angles. The test subjects were restrained with a three-point lap-shoulder belt and the Delta V was 30 km/h. RESULTS: The results from the Hybrid III and THOR 99 tests showed that, in most of the test, the head trajectories were an average of approximately 0.1 m shorter than those from equivalent PMHS. The Hybrid III and THOR 99 far-side belt geometry tests showed that the belt remained in place longer on the shoulder of the Hybrid III than on the THOR 99 and the THOR Alpha. This was probably due to a stiffer lumbar spine in the Hybrid III and to a large groove in the steel of the superior surface of the Hybrid III shoulder structure. The THOR 99 escaped from the shoulder belt about 40-50 ms earlier than the THOR Alpha. The results from the THOR Alpha tests show that the head trajectory accorded fairly well with the PMHS data, as long as the shoulder belt did not slip off the shoulder. Although the THOR Alpha shoulder escaped the shoulder belt in the 45 degrees far-side belt geometry, the PMHS did not. This may be due to the THOR Alpha shoulder design, with approximately 0.05 m smaller superior and medial shoulder range-of-motion, in combination with a relatively soft lumbar spine. CONCLUSIONS: The THOR Alpha provides head trajectories similar to those of the PMHS under these loading conditions, provided the shoulder belt remains in position on the shoulder. When the shoulder belt slipped off the dummy shoulder, the head kinematics was altered. The shoulder range-of-motion may be a contributing factor to the overall kinematics of an occupant in oblique frontal impact situations where the occupant moves in a trajectory at an angle from that of the longitudinal direction of the car. PMID- 16266944 TI - Field investigation of child restraints in side impact crashes. AB - OBJECTIVE: Various test procedures have been suggested for assessing the protection afforded by child restraints (CRS) in lateral collisions. Analyses of real world crashes can be used to identify relevant characteristics of the child, restraint, collision, and injury mechanisms that should be incorporated into the design of the test procedures as well as in the design of related ATDs and injury metrics. The objective of this work is to use in-depth crash investigations of children restrained in CRS in side impacts to elucidate specific sources and mechanisms of injuries and explore the role of crash severity variables such as magnitude and location of intrusion and specific impact angle. METHODS: Real world crashes involving children restrained in forward facing CRS in side impacts were analyzed from Partners for Child Passenger Safety, an on-going child specific crash surveillance system in which insurance claims are used to identify cases. In-depth crash investigations using standardized protocols were used to calculate the crash severity and determine the mechanisms and sources of the injuries sustained. RESULTS: Cases of 32 children restrained in CRS in 30 side impact crashes were examined. Twenty-five percent sustained AIS 2+ injuries. The most common injuries sustained by children restrained in CRS in side impact crashes were to the face, head, and lower extremity. Characteristics of the crashes that appeared related to injury were intrusion that entered the child's occupant space or caused an interior part of the vehicle to enter the child's occupant space, forward component of the crash, and the rotation of the CRS, restrained by a seat belt, towards the side of the impact. CONCLUSIONS: The ability to assess the injury potential in a laboratory setting for the body regions of common injury, the head, face, and lower extremity, must be explored. Characteristics of a regulatory-based test procedure to assess injury risk should include a frontal component to the crash and intrusion into the occupant's seating position. Design enhancements of the CRS should address rotation during lateral impacts. These results provide guidance to current efforts to design and regulate these restraints for the safety of child passengers in side impacts. PMID- 16266945 TI - Development of a three-year-old child FE model. AB - OBJECTIVE: Child crash dummies are conventionally used for safety performance evaluations of the child restraint system (CRS) in vehicle crash tests. To investigate injuries to various body regions of a child in detail, mathematical models are useful, and provide information that cannot be analyzed by crash dummies. Therefore, in the present research, a finite element (FE) model of a 3 year-old child has been developed by model-based scaling from the AM50 human FE model, THUMS (Total HUman body for Safety). METHODS: The dimensions of each body region were based on the anthropometry data of United States children, and material properties of child bone were estimated from data reported in the literature. Neck flexion, thorax impact responses, and torso flexion were validated against the response corridor of the 3-year-old Hybrid III dummy in calibration tests. A test of lap belt loading to the abdomen was also conducted. FE models of two different types of CRS, a 5-point harness and a tray shield CRS, were also made, and ECE R44 sled impact test simulations were conducted using the child FE model. RESULTS: The characteristics of the child FE model proved to be close to the Hybrid III and child volunteer corridor. In the ECE R44 sled test simulations using the child FE model, the head movement down and head rotation were large in the 5-point harness CRS, and chest deflection was large in the tray shield CRS. In both CRS types, the whole spine flexed in the child FE model. This behavior is different from that of the Hybrid III, where the thorax spine is stiff and only the cervical spine and lumbar spine flex. CONCLUSIONS: Although this child FE model has several limitations in areas such as the anatomical shapes and material properties of a child, this model can be a useful tool to examine the behavior of a child in impacts, which may be difficult to predict by using the Hybrid III dummy with its stiff thorax spine box. PMID- 16266946 TI - The role of door orientation on occupant injury in a nearside impact: a CIREN, MADYMO modeling and experimental study. AB - OBJECTIVE: This study addressed the effects of vehicle height mismatch in side impact crashes. A light truck or SUV tends to strike the door of a passenger car higher causing the upper border to lead into the occupant space. Conversely, an impact centered lower on the door, from a passenger car, causes the lower border to lead. We proposed the hypothesis that the type of injury sustained by the occupant could be related to door orientation during its intrusion into the passenger compartment. METHOD: Data on door orientation and nearside occupant injuries were collected from 125 side impact crashes reported in the CIREN database. Experimental testing was performed using a pendulum carrying a frame and a vehicle door, impacting against a USDOT SID. The frame allowed the door orientation to be changed. A model was developed in MADYMO (v 6.2) using the more biofidelic dummies, BIOSID, and SIDIIs as well as USDOT SID. RESULTS: In side impact crashes with the lower border of the door leading, 81% of occupants sustained pelvic injury, 42% suffered rib fractures, and the rate of organ injury was 0.84. With the upper border leading, 46% of occupants sustained pelvic injury, 71% sustained rib fracture, and the rate of organ injuries per case increased to 1.13. The differences in the groups with respect to pelvic injury were significant at p = 0.01, rib fracture, p = 0.10, and organ injury, p = 0.001. Experimental testing showed that when the door angle changed from lower to upper border leading, peak T4 acceleration increased by 273% and pelvic acceleration decreased by 44%. The model demonstrated that when the door angle changed from lower to upper border leading, the USDOT SID showed a 29% increase in T4 acceleration and a 57% decrease in pelvic acceleration. The BIOSID dummy demonstrated a 36% increase in T1 acceleration, a 44% increase in abdominal rib 1 deflection, a 91% increase in thoracic rib 1 deflection, and a 33% decrease in pelvic acceleration. CONCLUSIONS: These data add more insight to the problem of mismatch during side impacts, where the bumper of the striking vehicle overrides the door beam, causing the upper part of the door to lead the intrusion into the passenger compartment. Even with the same delta V and intrusion, with the upper border of the door leading, more severe chest and organ injuries resulted. This data suggests that door orientation should be considered when testing subsystems for side impact protection. PMID- 16266947 TI - Deflection, acceleration, and force corridors for small females in side impacts. AB - OBJECTIVE: This study was undertaken to develop biomechanical corridors applicable to the small-sized female in side impacts. METHODS: Sled tests were conducted using post mortem human subjects at a velocity of 6.7 m/s. Three chestbands were used to compute deflection-time histories at the axilla, xyphoid process, and tenth rib levels. Triaxial accelerometers were fixed to the upper and lower spine and sacrum to record acceleration-time histories. Specimens contacted the load wall with varying initial conditions (rigid and padded; flat wall and offset) from which impact forces to the thoracic, abdominal, and pelvic regions were obtained using load cell data. Adopting signal processing and mass based scaling methods, corridors were derived for forces, accelerations, and chest deflections at three levels for all initial conditions. RESULTS: All time history corridors were expressed as mean plus/minus one standard deviation and provided in the article. CONCLUSIONS: Acceleration-, deflection-, and force-time corridors obtained for the chest and pelvic regions of the human body will assist in the assessment of anthropomorphic test devices used in crashworthiness evaluations. PMID- 16266948 TI - Intervertebral neck injury criterion for prediction of multiplanar cervical spine injury due to side impacts. AB - OBJECTIVE: Intervertebral Neck Injury Criterion (IV-NIC) is based on the hypothesis that dynamic three-dimensional intervertebral motion beyond physiological limits may cause multiplanar injury of cervical spine soft tissues. Goals of this study, using a biofidelic whole human cervical spine model with muscle force replication and surrogate head in simulated side impacts, were to correlate IV-NIC with multiplanar injury and determine IV-NIC injury threshold for each intervertebral level. METHODS: Using a bench-top apparatus, side impacts were simulated at 3.5, 5, 6.5, and 8 g horizontal accelerations of the T1 vertebra. Pre- and post-impact flexibility testing in three-motion planes measured the soft tissue injury, i.e., significant increase (p < 0.05) in neutral zone (NZ) or range of motion (RoM) at any intervertebral level, above corresponding physiological limit. RESULTS: IV-NIC in left lateral bending correlated well with total lateral bending RoM (R = 0.61, P < 0.001) and NZ (R = 0.55, P < 0.001). Additionally, the same IV-NIC correlated well with left axial rotation RoM (R = 0.50, P < 0.001). IV-NIC injury thresholds (95% confidence limits) varied among intervertebral levels and ranged between 1.5 (0.6-2.4) at C3 C4 and 4.0 (2.4-5.7) at C7-T1. IV-NIC injury threshold times were attained beginning at 84.5 ms following impact. CONCLUSIONS: Present results suggest that IV-NIC is an effective tool for determining multiplanar soft tissue neck injuries by identifying the intervertebral level, mode, time, and severity of injury. PMID- 16266949 TI - The use of seat belt by motor vehicle occupants in South Africa. AB - OBJECTIVE: The purpose of this article was to examine the use of seat belt by motor vehicle users in the various provinces in South Africa. METHODS: Data were abstracted from published reports of the national Department of Transport. Percentage distribution and correlation of road safety variables and seat belt wearing rates for motor vehicle users were calculated for the different provinces. RESULTS: High seat belt wearing rates by the drivers (between 75.1% and 88.1%, national rate was 81%). The seat belt wearing rates for the front and back seat passengers were much lower than for the drivers. The seat belt wearing rates for front seat passengers (44.5% to 60.5%, national - 50.1%) and back seat passengers (1% to 16%, national - 7.6%) were much lower than for the drivers. The national seat belt wearing rate for all vehicles between 1982 and 1995 was between 46.9% and 69.2%, but this has generally declined. CONCLUSION: There is a need for the implementation of strategies to increase the use of seat belts to reduce injuries and fatalities. PMID- 16266950 TI - Advances in the management of pediatric asthma: a review of recent FDA drug approvals and label updates. AB - Children have the highest prevalence of asthma of any age group. In the United States during 2001, there were 12.6 million physician and hospital outpatient visits for asthma treatment, of which almost 5 million involved children 18 years and younger. Therapeutic advances in pediatric asthma could improve patient outcomes and potentially reduce the burden on health care systems. Efforts to obtain efficacy and safety data in pediatric populations and develop pediatric formulations of asthma treatments have been encouraged by the FDA and clinicians. This article reviews the newest additions to asthma therapies approved for use in children, including an inhaled corticosteroid, some long-acting beta2-agonists, some leukotriene-receptor blockers, and a single-isomer, short-acting beta2 agonist. PMID- 16266951 TI - Contrast of pediatric asthma management approaches in a multicultural and collectivistic population. AB - BACKGROUND: Although asthma is the most prevalent chronic illness in childhood and affects approximately 9 million children, the management approaches used by practitioners may not be efficient from the perspective of families. Clinicians often maintain their usual customs of practice, and the context of the clinical encounter is defined in terms of an individual illness management. In collectivistic and multiethnic settings, the extended kin group or extended family shares responsibility for illness management. The goal of this study is to describe health care providers' strategies to manage children with asthma in a multicultural and collectivistic cultural context. METHODS: Data were obtained through semistructured in-depth practitioner interviews. Western-trained and traditional practitioners participated. RESULTS: Narrative analysis strategies produced two major themes: 1) fix the asthma and 2) making connections. Practitioners who perceived their responsibility to fix the asthma (make things physiologically normal) often ran into constraints with dealing with the extended family group. Other practitioners who used strategies of making connections often capitalized on the assistance of others to problem-solve the asthma management. CONCLUSION: In terms of asthma management, the barriers most frequently reported by practitioners were related to the sociocultural and physical environment. There may be vast differences in asthma management approaches for populations from collectivistic cultural orientation. PMID- 16266952 TI - Asthma patients' knowledge, perception, and adherence to the asthma guidelines. AB - Asthma patients' knowledge and perceptions about asthma management regimens have been shown to correlate with the quality of their asthma management and outcomes. This study addresses adult ambulatory asthma patients' knowledge and perception of the asthma management guidelines issued by the National Asthma Education and Prevention Program. We interviewed 76 patients at three adult medicine outpatient clinics. Of 37 patients who knew about peak flow monitoring, 21 found it to be useful, only 10 were using it regularly, and 7 had a notation about it in their chart. Eight patients knew about patient self-assessment forms, 6 found them useful, 3 had used one recently, and only 1 patient had one in the chart. Six patients knew about asthma action plans, 4 of them thought that these plans were useful, but only 1 patient was actively using it. This study demonstrates major gaps in the asthma patients knowledge, perceptions, and their adherence to recommendations and also with the documentation in the medical record. PMID- 16266953 TI - Successful treatment of a patient with severe Churg-Strauss syndrome by a combination of pulse corticosteroids, pulse cyclophosphamide, and high-dose intravenous immunoglobulin. AB - A 24-year-old woman with a 4-year history of bronchial asthma suffered from bloody sputum, numbness of the extremities, elevated eosinophil count, and hypoxemia. A diagnosis of alveolar hemorrhage was made by bronchoalveolar lavage. Echocardiogram revealed severe hypokinesis of the left ventricular wall. Her respiratory condition deteriorated despite administration of pulse corticosteroids. A second pulse corticosteroids and pulse cyclophosphamide followed by high-dose intravenous immunoglobulin brought about a dramatic improvement of alveolar hemorrhage, cardiac impairment, and peripheral neuropathy. Levels of antimyeloperoxidase-antineutrophil cytoplasmic antibodies, soluble thrombomodulin, soluble interleukin-2 receptor, eosinophil cationic protein were elevated and returned to the normal range in remission. The combination of pulse corticosteroids, pulse cyclophosphamide, and high-dose intravenous immunoglobulin seemed effective for the acute phase of severe Churg Strauss syndrome. PMID- 16266954 TI - Prevalence of probable mental disorders among pediatric asthma patients in an inner-city clinic. AB - OBJECTIVE: To determine the screen-positive prevalence of anxiety disorders and depression among pediatric asthma patients in an inner-city asthma clinic and to investigate the association between probable diagnoses of anxiety disorders and depression and medical service use among inner-city pediatric asthma patients. METHOD: In this pilot study, a consecutive sample of pediatric asthma patients aged 5-11 in the waiting room of an inner-city asthma clinic was screened for mental disorders using the DISC Predictive Scales (DPS), which produces probable DSM-IV diagnoses. In addition, data on health service use for asthma were collected. Statistical analyses were performed to examine the relationship between probable anxiety disorders and depression and health service use for asthma among pediatric asthma patients. RESULTS: Approximately one in four (25.7%) pediatric asthma patients in an inner-city asthma clinic met criteria for a probable diagnosis of current anxiety disorders or depression (past 4-week prevalence). Specifically, childhood separation anxiety disorder was common among 8.1%, panic among 14.9%, generalized anxiety disorder among 4.1%, agoraphobia among 5.4%, and 2.7% had depression. Having more than one anxiety disorder or depression diagnosis was associated with higher levels of inpatient and outpatient medical services, compared with patients who were negative on screening for anxiety or depressive disorders, although differences failed to reach statistical significance. CONCLUSIONS: These findings are the first to provide preliminary evidence suggesting that mental health problems are common among pediatric asthma patients in an inner-city clinic. The results also suggest that mental health problems in pediatric asthma patients may be associated with elevated levels of medical service use for asthma. Replication of this pilot study is needed with a larger sample, more precise diagnostic methodology, and a comparison group with chronic medical illness. PMID- 16266955 TI - Halothane, an effective infrequently used drug, in the treatment of pediatric status asthmaticus: a case report. AB - Asthma is the most common chronic disease of childhood. Despite a better understanding of the disease process and its management, status asthmaticus continues to be a life-threatening event. The use of volatile inhaled anesthetics is infrequently reported as adjunctive therapy to conventional treatment of this condition. We report the use of halothane in a mechanically ventilated pediatric patient with life-threatening status asthmaticus who was admitted to the pediatric intensive care unit (PICU) after failing to respond to standard medical therapy and noninvasive positive pressure ventilation. A 12-year-old African American male was seen in the emergency department and treated with intravenous corticosteroids, beta-agonist therapy. He deteriorated rapidly and required endotracheal intubation and mechanical ventilation. Two hours later, the patient developed an acute, severe respiratory acidosis (pH=6.97, PaCO2=171, PaO2=162, BE=1.7). Halothane was started at 2% by using the Siemens Servo 900C anesthesia ventilator. Improvement in both arterial blood gases and exhaled tidal volume were noted 30 minutes after initiation of the anesthetic gas. The patient remained on halothane for a total of 36 hours. No adverse effects associated with the use of halothane were noted. The patient was extubated to BiPAP 16/6, FiO2=0.30 at 68 hours and was discharged home 5 days later. PMID- 16266956 TI - Childhood asthma hospitalization rates, childhood asthma prevalence, and their relationships in Erie County, New York. AB - We estimated asthma hospitalization rates, prevalence, and their relationships for children under age 15 in Erie County, New York. Information on hospitalizations was obtained from hospital discharge data, and prevalence was estimated through a mailed survey of 9271 children. The hospitalization rate was 2.3 per 1000, the lifetime prevalence was 14.1%, and the hospitalization-to prevalence ratio was 16.3 per 1000 prevalent cases. Hospitalization rates and hospitalization-to-prevalence ratios were higher among young children relative to older children; and hospitalization rates, prevalence, and hospitalization-to prevalence ratios were higher among males, blacks, and Buffalo residents, suggesting greater severity, poorer management, and/or less access to care among these groups. The hospitalization-to-prevalence ratio may be useful in estimating prevalence from more easily available hospitalization data. PMID- 16266957 TI - Factors associated with asthma control. AB - The aim of this study was to evaluate which factors are associated with asthma control experienced by asthma patients. In a cross-sectional study patients aged 16-60 years with mild to moderate asthma were selected. The influence of the following factors on asthma control was studied in a multivariate model: age, gender, socioeconomic status, smoking, perceived hyperresponsiveness (PHR, responding with asthma symptoms to one or more triggers), allergy (Phadiatop), long-acting bronchodilating agents, and inhaled corticosteroids. Asthma control was measured by means of the Asthma Control Questionnaire (ACQ) as developed by Juniper. Forced expiratory volume in 1 second (FEV1) was measured by means of a portable spirometer. In this study with 311 patients, mean ACQ score was 1.39 (range 0-4.43). A stepwise backward linear regression analysis showed that low socioeconomic status (beta 0.425; p=0.001), current smoking (beta 0.555; p<0.001), high dose of inhaled corticosteroids (beta 0.364; p=0.04) and perceived hyperresponsiveness for increasing number of different triggers (PHR for 1 trigger beta 0.833; p=0.03; 2 triggers beta 0.810; p=0.03; 3 triggers beta 0.995; p=0.01; 4 triggers beta 1.131; p=0.002; 5 triggers beta 1.182; p=0.002) are independent predictors for poorer asthma control. Beside treatment with medication, stopping smoking and avoidance of triggers are factors, which may have a high impact on asthma control. PMID- 16266958 TI - Exhaled nitric oxide and bronchial wall thickening in asthmatics during and after acute exacerbation: evidence of bronchial wall remodeling. AB - To assess whether bronchial wall thickening during asthma exacerbations is due to active inflammation in severe asthmatics, we measured bronchial wall thickness and exhaled nitric oxide (FeNO) following treatment. Nine asthmatics were compared with seven controls with high-resolution computed tomography, spirometry, and FeNO measurements. The asthmatic bronchial wall area percent and FeNO was greater than controls. Following treatment, the FEV1 markedly improved, FeNO decreased modestly, and bronchial wall area percent did not change significantly. Bronchial wall thickening persisted after treatment of acute asthma exacerbation despite improvement in spirometry and decline in FeNO, possibly due to chronic airway remodeling. PMID- 16266959 TI - Participatory decision making, asthma action plans, and use of asthma medication: a population survey. AB - Use of controller asthma medication and possession of asthma action plans remains suboptimal. Our aim was to investigate the association of the propensity of physicians to involve patients in their care (participatory decision-making style) and their asthma management in a representative population sample of 3015 adults. Current doctor-diagnosed asthma was reported by 393 (13.0%). People who rated their doctors as more participatory were significantly more likely to report more regular use of controller medications and possession of a written asthma action plan, but not less asthma morbidity. Possession of a written action plan was associated with more participatory interactions (OR 2.3; 95% CI 1.1-4.7, for upper tertile scores compared to lowest tertile); more severe symptoms (OR 4.8; 95% CI 1.7-13.0), being female (OR 2.2; 95% CI 1.2-4.3), those with higher education, and residence outside the metropolitan area (OR 2.1; 95% CI 1.1-4.0). Increasing patient participation in their own care is associated with better asthma management, independent of asthma symptoms. Longitudinal studies are needed to examine if increasing participation in decisions can also improve asthma outcomes. PMID- 16266960 TI - The emergency department as an asthma surveillance tool at the community level: a decline in the burden of pediatric asthma in halifax, Canada. AB - We report pediatric emergency department (PED) asthma visit and inpatient asthma (AS) admission data in our area over a 5-year period. AS visits decreased by 33.9%, AS admissions by 24.6%, both significant compared with the decline in elementary school enrollment. The decrease in asthma visits was due to a decrease in the number of asthmatic patients, not a decline in repeat visits, or use of alternate venues of care. Explanations include a decrease in the burden of disease or an improvement in ambulatory care, but not alternate treatment venues or improvement in acute (PED) care. Readily available, emergency department data are useful in the community surveillance of asthma. PMID- 16266961 TI - Learning preferences of caregivers of asthmatic children. AB - BACKGROUND: People learn in different ways: visually, aurally, by reading/writing, and kinesthetically. In our clinic, we use color-coded Asthma Action Cards to educate our patients and their caregivers on asthma management. Our teaching is largely aural based, with the cards providing reading and visual stimulation and hands-on practice with devices offering kinesthetic stimulation. OBJECTIVE: We sought to determine the learning styles of the caregivers of our asthmatic children. METHODS: Caregivers in our Asthma/Allergy Clinic completed the Visual-Aural-Read/Write-Kinesthetic (VARK) questionnaire anonymously, and the responses were evaluated on the basis of previously validated scoring instructions. RESULTS: Analysis of 98 respondents showed that 42% had a single learning modality preference, and the remaining 58% were multimodal learners. Of those who reported a single mode of learning, 61% preferred kinesthetic, 27% preferred reading/writing, and less than 1% each preferred aural or visual stimuli. Of all 98 caregivers, 82% included kinesthetic as a learning preference, 59% included read/write, 50% included aural, and 41% included visual. CONCLUSION: The majority of caregivers preferred the kinesthetic learning method, whether as a single learning preference or in combination with other approaches. Incorporating kinesthetic methods of learning, such as role plays and problem solving case scenarios, into standardized asthma education curricula may be beneficial to patients and families in terms of understanding and using their regimen. PMID- 16266962 TI - Efficacy and tolerability of budesonide Clickhaler and Turbuhaler in adult asthma. AB - New dry powder inhalers should be clinically comparable with established devices to ensure the continuity of effective therapy for asthma patients. This randomized, open, parallel group study compared the clinical efficacy and tolerability of budesonide delivered via Clickhaler or Turbuhaler dry powder inhalers in adults with mild to moderate stable asthma. Following a 4-week stabilizing period using budesonide Turbuhaler adults aged 18 years or older, who had been treated with inhaled corticosteroids for at least the previous 12 weeks, were randomized to receive budesonide twice daily (or=85 percentile), obese girls (BMI>or=95 percentile) had an increased risk of current wheezing of 19% (OR=1.19; 95% CI 0.97-1.46). After stratifying by age at menarche, we observed that this increased risk was only present in girls with menarche at 11 years old or younger (1.31%; 95% CI 1.01-1.73). CONCLUSIONS: The association between obesity and asthma seems to be greater among girls with early puberty, suggesting the role of female hormones. PMID- 16266965 TI - Between morality and repentance: recapturing "sin" for bioethics. AB - Distinguishing within "sin" the dimensions of anomia, hamartia, and asthenia makes it possible to analyze in greater detail the contrary manners in which traditional and post-traditional Christianities in this issue of Christian Bioethics endeavor to recapture what was lost when secular bioethics reconstructed the specifically spiritual-context-oriented normative commitments of Christianity in one-dimensionally moral terms. Various post-traditional attempts at securing moral orientation and resources for forgiveness, both of which secular bioethics finds increasingly difficult to provide, are critically reviewed. Their engagement of secular moral concepts and concerns, and even their adoption of an academically philosophical posture and language, is presented as responsible for their failure to adequately preserve what in traditional Christianity would count as prohibited vs. permitted, and advisable vs. non advisable, or what would allow to resolve "tragic conflicts." The deeper reason for this failure lies in post-traditional Christianity's restricting the Christian life (with its central tension between love and the law) to what can be captured by cognitive categories. As the survey of several traditionally Christian accounts of sin in bioethics makes clear, both moral orientation (along with the resolution of "tragic" conflicts) and the sources of forgiveness are available, once that Christian life is framed in terms of persons' spirit supported practical involvement in ascesis and liturgy, and once bioethical reflections are situated in the experiential context of such involvement. PMID- 16266966 TI - Sin and bioethics. AB - The essay starts out with defining the biblical concept of sin in the Old and the New Testaments. The literal knowledge of divine truth is distinguished from its truthful and spiritual interpretation. A further distinction should be made between the Creator of life (God) and the medium or "intermediary creator" (man) of life. I argue for the "single wholeness" of the human race and for the unity of human responsibility in bioethics. In delineating the teaching of the Church on abortion and family planning, I show that the healing of all human diseases, from traditional interventions to genetic ones, is a Christian duty and is in accordance with Christ's mission on earth as long as one has not been directly or indirectly involved in "reproducing" or "designing" one's descendants or destroying or damaging human life even at its very beginnings. PMID- 16266967 TI - Freedom in responsibility: on the relevance of "sin" as a hermeneutic guiding principle in bioethical decision making. AB - This essay deals with questions of responsibility concerning technology, in particular, gene technology and the special problem of research on embryos. I raise issues concerning the extent of humans' authority to act and the limits of human freedom. In what way is that freedom given, and what kind of responsibility results from it? By discussing various concepts of human freedom in the tradition of European philosophy, as juxtaposed to the Protestant understanding of freedom, this essay discusses the restricting limits, and the obligation to take responsibility. It will turn out that the question concerning freedom cannot be answered without understanding what being human involves. From a Christian perspective, this implies that the foundational relationship between human freedom and sin will be central to an assessment of the human ability to take responsibility. By obliterating the limits of human freedom, sin jeopardizes the very essence of that freedom. The project of taking into account the sinful state of the human condition thus aims at developing a realistic picture of the authority of humans in action, even in view of the human tasks of promoting science and research. PMID- 16266968 TI - Freedom in responsibility: a response. AB - This paper is a critical response to Elisabeth Grab-Schmidt's article "Freedom in responsibility: On the relevance of 'sin' as hermeneutic guiding principle in bioethical decision making." Grab-Schmidt's chief contention is that ethics begins with anthropology, and that moral responsibility is thereby grounded within a set of given limits. Freedom is distorted into sin when these limits are transgressed. My principal complaint is that her account of the relationship between freedom and sin is grounded in a tragic ontology. Alternatively, I contend that anthropology is grounded in Christology in which freedom is a gift of the Spirit. Consequently, sin is not so much tragic as it marks a refusal of humans to accept their divine election. The issues of human cloning and embryonic stem-cell research are used to exemplify what difference these respective differences might make in a process of moral deliberation. PMID- 16266969 TI - Bioethics and sin. AB - On the basis of a historical reconstruction of the stages through which the Christian notion of sin took shape in Protestantism, the significance of this term for modern bioethics is derived from its opposition to a holiness of God and his creatures, which in turn translates into the secular moral concept of dignity. This dignity imposes obligations to respect and to relationships that are sustained by faithfulness and trust. In being based on the gratuitousness of God's grace, such relationships preclude attempts at instrumentalization, denial of singularity, and subjection to market forces. Accordingly, reproductive cloning as well as exposing medicine to economical considerations can be classified as sinful. The difference between sinful acts and humans' sinful state furthermore permits to address the problems of evil and misfortune in the world, and to acknowledge humans' responsibility for the threats to humanity entailed by those ills. While the Christian faith relies on God's mercy, it also imposes the task of following Christ by fighting against evil and misfortune. PMID- 16266970 TI - Notes on "bioethics and sin" by Jean-Francois Collange. AB - Placing the notion of sin in the context of a meontic account of evil, and emphasizing the effect of sin on the sinner himself, this commentary exposes the insufficiency of restricting oneself to human efforts at atonement, and of thus underemphasizing the role of Christ. Collange's claim that the teaching of "predestination" is rooted in Paul and that the doctrine of merits and indulgences is rooted in Augustine is criticized, and Luther's "forensic" understanding is linked with Augustine, rather than with Paul. Collange's reduction of the concern for holiness to respect and trust is contrasted with holiness's essential context of loving unification with God. The commentary closes by exposing the unsatisfactory scantiness of Collange's treatment of cloning, health-care economy, and of the evils of life. PMID- 16266971 TI - The significance of the concept of sin for bioethics. AB - After a period during which the theological categories of sin and forgiveness were ignored or trivialized, presently these notions are being rediscovered. What could their impact be on bioethics, either in the narrow sense of medical ethics, or in the more encompassing sense of the ethics of the life sciences? This essay begins with describing the processes of transcending and ethitization, which gave rise to the biblical notion of sin. It portrays the theological foundation of sin in terms of a twofold refusal of proper relations to God and other humans. Through the practise of confession in the face of God (coram deo), sin is placed into a horizon of hope for forgiveness and reconciliation. The heuristic and hermeneutical significance of these categories results from their introducing a "surplus value," which transcends biological and ethical considerations. This additional dimension is illustrated in view of care (cura) for the injured, and in view of individual as well as collective willingness to forgive. PMID- 16266972 TI - "... As we forgive those who trespass against us...": theological reflections on sin and guilt in the hospital environment. AB - In general parlance the term sin has lost its existential meaning. Originally a Jewish-Christian term within a purely religious context, referring to a wrongdoing with regard to God, sin has slowly become reduced to guilt in the course of the secularization process. Guilt refers to a wrongdoing, especially with regard to fellow human beings. It also refers to errors of judgement with what can be tragic consequences. These errors can occur whenever human beings are called upon to act, including the hospital environment. A Christian hospital has to address the issue of how to deal not only with guilt-ridden misdemeanors, but also with wrongdoing unto God, which overshadows every instance of guilt-ridden human behavior. Here, as in every parish, the Church Service is the place to acknowledge sin, confess sin, and forgive sin, beyond the boundaries of the parish itself. PMID- 16266973 TI - Sin and bioethics: why a liturgical anthropology is foundational. AB - The project of articulating a coherent, canonical, content-full, secular morality cum-bioethics fails, because it does not acknowledge sin, which is to say, it does not acknowledge the centrality of holiness, which is essential to a non distorted understanding of human existence and of morality. Secular morality cannot establish a particular moral content, the harmony of the good and the right, or the necessary precedence of morality over prudence, because such is possible only in terms of an ultimate point of reference: God. The necessity of a rightly ordered appreciation of God places centrally the focus on holiness and the avoidance of sin. Because the cardinal relationship of creatures to their Creator is worship, and because the cardinal corporate act of human worship is the Liturgy, morality in general and bioethics in particular can be understood in terms of the conditions necessary, so as worthily to enter into Eucharistic liturgical participation. Morality can be summed up in terms of the requirements of ritual purity. A liturgical anthropology is foundational to an account of the content-full morality and bioethics that should bind humans, since humans are first and foremost creatures obliged to join in rightly ordered worship of their Creator. When humans worship correctly, when they avoid sin and pursue holiness, they participate in restoring created reality. PMID- 16266974 TI - Fas receptor signaling is requisite for B cell differentiation. AB - The Fas/Fas ligand (FasL) pathway has been largely implicated in the homeostasis of mature cells. However, it is still unclear whether it plays a role at the progenitor level. To address this issue, we created chimeric mice by transferring C57BL/6 bone marrow (BM) cells of the lpr (Fas-FasL+) or gld (Fas+FasL-) genotype into Rag-2-/- hosts of the same genetic background. In this model, the consequences of a deficient Fas/FasL pathway on lymphoid differentiation could be evaluated without endogenous competition. Analysis of the chimerism revealed a differential sensitivity of hematopoietic lineages to the lack of Fas receptor signaling. While donor-derived myelo-monocytic cells were similarly distributed in all chimeric mice, mature B cells were deleted in the BM and the spleen of lpr chimera, leading to the absence of the marginal zone (MZ) as detected by immunohistology. In contrast, B cell hematopoiesis was complete in gld chimera but MZ macrophages undetectable. These defects suggest a direct and determinant dual role of FasL regulation in negative selection of B cells and in maintenance of the MZ. PMID- 16266975 TI - Claudin proteins in human cancer: promising new targets for diagnosis and therapy. AB - The tight junction proteins claudins are abnormally regulated in several human cancers. In particular, claudin-3 and claudin-4 are frequently overexpressed in several neoplasias, including ovarian, breast, pancreatic, and prostate cancers. Although the exact roles of these proteins in tumorigenesis are still being uncovered, it is clear that they represent promising targets for cancer detection, diagnosis, and therapy. PMID- 16266976 TI - Hopping around the tumor genome: transposons for cancer gene discovery. AB - Retroviruses are powerful insertional somatic mutagens that have been used for many landmark discoveries of cancer genes in model organisms. However, their use as a cancer gene discovery tool has been limited to only a few tissues, mainly the hematopoietic system and mammary gland. Recently, the Sleeping Beauty (SB) transposon system was shown to be useful for random somatic cell mutagenesis in mice, allowing the induction or acceleration of tumor formation both in the hematopoietic system and in sarcomas. In these tumors, SB transposons repeatedly "tagged" specific genes, both known and new cancer genes. These results indicate that the SB system has great potential both for generating specific mouse models of human cancer and for cancer gene discovery in a wide variety of tissues. PMID- 16266977 TI - Antiandrogen bicalutamide promotes tumor growth in a novel androgen-dependent prostate cancer xenograft model derived from a bicalutamide-treated patient. AB - Androgen ablation therapies are effective in controlling prostate cancer. Although most cancers relapse and progress despite androgen ablation, some patients experience antiandrogen withdrawal syndrome, in which those treated with antiandrogen show clinical improvement when antiandrogen is discontinued. Although the androgen receptor (AR) is suggested to play an important role in prostate cancer progression even after the androgen ablation, limited tissue availability for molecular studies and small numbers of human prostate cancer cell lines have restricted prostate cancer research. Here, we describe KUCaP, a novel serially transplantable human prostate cancer xenograft model. We established KUCaP from liver metastatic tissue of a patient treated with antiandrogen bicalutamide. KUCaP expressed the AR with a point mutation at amino acid 741 (tryptophan to cysteine; W741C) in the ligand-binding domain. This mutation was also present in cancerous tissue used for generation of KUCaP. Although the growth of KUCaP in male mice was androgen dependent, bicalutamide aberrantly promoted the growth and prostate-specific antigen production of KUCaP. For the first time, we show the agonistic effect of bicalutamide to a xenograft with clinically induced AR mutation. This bicalutamide-responsive mutant AR will serve in the development of new therapies for androgen ablation-resistant prostate cancers. PMID- 16266978 TI - Adenovirus-mediated overexpression of REIC/Dkk-3 selectively induces apoptosis in human prostate cancer cells through activation of c-Jun-NH2-kinase. AB - Alteration in genes which takes place during malignant conversion and progression could be potential targets for gene therapy. We previously identified REIC/Dkk-3 as a gene whose expression is reduced in many human cancers. Here, we showed that expression of REIC/Dkk-3 was consistently reduced in human prostate cancer tissues in a stage-dependent manner. Forced expression of REIC/Dkk-3 induced apoptosis in human prostate cancer cell lines lacking endogenous REIC/Dkk-3 expression but not in REIC/Dkk-3-proficient normal prostate epithelial and stromal cells. The apoptosis involved c-Jun-NH2-kinase activation, mitochondrial translocation of Bax, and reduction of Bcl-2. A single injection of an adenovirus vector carrying REIC/Dkk-3 showed a dramatic antitumor effect on a xenotransplanted human prostate cancer. Thus, REIC/Dkk-3 could be a novel target for gene-based therapy of prostate cancer. PMID- 16266979 TI - Telomerase expression in noncancerous bronchial epithelia is a possible marker of early development of lung cancer. AB - Centrally located lung cancers in smokers frequently associated with subsequent primary tumors. We evaluated the telomerase expression chronologically in noncancerous epithelia as a risk factor of susceptibility to lung cancer development. Telomerase protein expression was examined in situ by immunohistochemistry in 26 noncancerous bronchial epithelia adjacent to centrally located early-stage lung cancers in sequential 23 patients treated by photodynamic therapy or surgery among 206 patients who underwent autofluorescence bronchoscopy from 1997 to 2003. Among the 15 lesions in 12 patients treated by photodynamic therapy alone, 11 lesions achieved complete remission after photodynamic therapy, and none of their noncancerous bronchial epithelia was telomerase positive. On the contrary, in the remaining four lesions, either recurrence or secondary lung cancer developed adjacent to the successfully treated primary cancer within 26 months, and the telomerase protein expression in noncancerous epithelia was detected before the secondary cancer development (P < 0.001). The overall relationship of human telomerase reverse transcriptase positivity in noncancerous epithelia and subsequent lung cancer development, including patients treated by radiation or surgery, showed higher significance (P < 0.0001). Histologically "normal" bronchial epithelia in smokers may unphysiologically express telomerase as a field, and such epithelia are likely susceptible to develop lung cancer. We propose that ectopic expression of telomerase in bronchial epithelia may precede transformation in human lung cancer development and that detection of telomerase protein in noncancerous bronchial epithelia will become a useful marker detecting high-risk patients for lung cancer development. PMID- 16266980 TI - A polycistronic microRNA cluster, miR-17-92, is overexpressed in human lung cancers and enhances cell proliferation. AB - MicroRNAs (miRNAs) are small noncoding RNAs, thought to be involved in physiologic and developmental processes by negatively regulating expression of target genes. We have previously reported frequent down-regulation of the let-7 miRNA family in lung cancers and, in the present study, assessed alteration in a panel of 19 lung cancer cell lines. As a result, we found for the first time that the miR-17-92 cluster, which comprises seven miRNAs and resides in intron 3 of the C13orf25 gene at 13q31.3, is markedly overexpressed in lung cancers, especially with small-cell lung cancer histology. Southern blot analysis revealed the presence of increased gene copy numbers of the miRNA cluster in a fraction of lung cancer cell lines with overexpression. In addition, we were able to show predominant localization of C13orf25 transcripts within the nucleus and introduction of the expression construct of the miR-17-92 cluster, but not the putative open reading frame of C13orf25, enhancing lung cancer cell growth. These findings clearly suggest that marked overexpression of the miR-17-92 cluster with occasional gene amplification may play a role in the development of lung cancers, especially in their most aggressive form, small-cell lung cancer, and that the C13orf25 gene may well be serving as a vehicle in this regard. PMID- 16266982 TI - Fine mapping reveals multiple loci and a possible epistatic interaction within the mammary carcinoma susceptibility quantitative trait locus, Mcs5. AB - To identify high-frequency, low-penetrance breast cancer modifier genes, we have developed a rat genetic model that uses the Wistar-Kyoto (WKy) inbred strain, resistant to developing 7,12-dimethylbenz[a]anthracene-induced mammary carcinogenesis, as a congenic donor and the susceptible Wistar-Furth (WF) strain as the recipient. Here, data from congenic rat lines containing smaller WKy genomic intervals of the Mcs5 quantitative trait locus region are presented to fine map three independently acting Mcs5 subloci. WKy-homozygous females from congenic lines defining Mcs5a, Mcs5b, and Mcs5c averaged, respectively, 4.0 +/- 0.4, 11.6 +/- 0.6, and 3.5 +/- 0.4 mammary carcinomas per rat. These phenotypic values are statistically different from the WF-homozygous phenotype value of 8.0 +/- 0.4, which is the baseline phenotype used for these experiments. We identified a likely Mcs5a x Mcs5b epistatic interaction that results in masking the increased susceptibility effect of the Mcs5b WKy allele by the Mcs5a WKy allele. We also provide evidence for a Mcs5a x Mcs5c interaction that is synergistic to decrease mammary carcinoma susceptibility below the additive effects of WKy alleles at each locus independently. The Mcs5 subloci are currently localized to 1.0, 7.5, and 4.5 Mb of rat chromosome 5, and the orthologous regions are on human chromosome 9 and mouse chromosome 4. These loci will provide unbiased candidate gene loci for evaluation in human case-control association studies. PMID- 16266981 TI - An in vivo tumor model exploiting metabolic response as a biomarker for targeted drug development. AB - In vivo models that recapitulate oncogene-dependent tumorigenesis will greatly facilitate development of molecularly targeted anticancer therapies. We have developed a model based on activating mutations in c-KIT in gastrointestinal stromal tumors (GISTs). This model comprises murine tumors of FDC-P1 cell lines expressing c-KIT mutations that render the tumors either responsive (V560G) or resistant (D816V) to the small-molecule c-KIT inhibitor, imatinib. Clinically, GIST response to imatinib is associated with rapid reduction in fluorodeoxyglucose (FDG) uptake on positron emission tomography (PET), preceding changes in conventional response criteria by several weeks. Using the FDC-P1 model in small animal PET, FDG uptake into tumors expressing the c-KIT V560G mutation was significantly reduced as early as 4 hours after imatinib treatment. In contrast, no change in FDG uptake was observed in resistant c-KIT D816V expressing tumors after 48 hours of imatinib treatment. Consistent with the PET results, expression of the glucose transporter, GLUT1, was significantly reduced in V560G tumors at 4 hours, preceding changes in markers of proliferation by several hours. In vitro, imatinib treatment of V560G cells resulted in a reduction of glucose transporter numbers at the cell surface and decreased glucose uptake well before changes in cell viability. Notably, decreased ambient glucose concentrations enhanced the cytotoxic effect of imatinib. Taken together, these data account for the rapidity and significance of the PET response to imatinib and suggest that metabolic effects may contribute to imatinib cytotoxicity. Further, the FDC-P1 model represents a very useful paradigm for molecularly targeted drug development. PMID- 16266983 TI - Constitutive activation of Akt by Flt3 internal tandem duplications is necessary for increased survival, proliferation, and myeloid transformation. AB - Up to 30% of patients with acute myeloid leukemia (AML) harbor internal tandem duplications (ITD) within the FLT3 gene, encoding a receptor tyrosine kinase. These mutations induce constitutive tyrosine kinase activity in the absence of the natural Flt3 ligand and confer growth factor independence, increased proliferation, and survival to myeloid precursor cells. The signaling pathways and downstream nuclear targets mediating leukemic transformation are only partly identified. Here, we show that the presence of Flt3-ITD constitutively activates Akt (PKB), a key serine-threonine kinase within the phosphatidylinositol 3-kinase pathway. Constitutive activation of Akt phosphorylated and inhibited the transcription factor Foxo3a. Restored Foxo3a activity reversed Flt3-ITD-mediated growth properties and dominant-negative Akt prevented Flt3-ITD-mediated cytokine independence. Conditional Akt activation targeted to the cell membrane induced cytokine-independent survival, cell cycle progression, and proliferation. Importantly, Akt activation was sufficient to cause in vitro transformation of 32D myeloid progenitor cells and in vivo promoted the development of a leukemia like myeloid disease. Akt phosphorylation was found in myeloid blasts of 86% of AML patients, suggesting an important role in leukemogenesis. In summary, Akt is necessary for increased survival, proliferation, and leukemic transformation by Flt3-ITD, possibly by inactivation of Foxo transcription factors. These findings indicate that Akt and Foxo transcription factors are attractive targets for therapeutic intervention in AML. PMID- 16266984 TI - Novel pheochromocytoma susceptibility loci identified by integrative genomics. AB - Pheochromocytomas are catecholamine-secreting tumors that result from mutations of at least six different genes as components of distinct autosomal dominant disorders. However, there remain familial occurrences of pheochromocytoma without a known genetic defect. We describe here a familial pheochromocytoma syndrome consistent with digenic inheritance identified through a combination of global genomics strategies. Multipoint parametric linkage analysis revealed identical LOD scores of 2.97 for chromosome 2cen and 16p13 loci. A two-locus parametric linkage analysis produced maximum LOD score of 5.16 under a double recessive multiplicative model, suggesting that both loci are required to develop the disease. Allele-specific loss of heterozygosity (LOH) was detected only at the chromosome 2 locus in all tumors from this family, consistent with a tumor suppressor gene. Four additional pheochromocytomas with a similar genetic pattern were identified through transcription profiling and helped refine the chromosome 2 locus. High-density LOH mapping with single nucleotide polymorphism-based array identified a total of 18 of 62 pheochromocytomas with LOH within the chromosome 2 region, which further narrowed down the locus to <2 cM. This finding provides evidence for two novel susceptibility loci for pheochromocytoma and adds a recessive digenic trait to the increasingly broad genetic heterogeneity of these tumors. Similarly, complex traits may also be involved in other familial cancer syndromes. PMID- 16266985 TI - HIN-1, an inhibitor of cell growth, invasion, and AKT activation. AB - The HIN-1 gene encoding a small, secreted protein is silenced due to methylation in a substantial fraction of breast, prostate, lung, and pancreatic carcinomas, suggesting a potential tumor suppressor function. The receptor of HIN-1 is unknown, but ligand-binding studies indicate the presence of high-affinity cell surface HIN-1 binding on epithelial cells. Here, we report that HIN-1 is a potent inhibitor of anchorage-dependent and anchorage-independent cell growth, cell migration, and invasion. Expression of HIN-1 in synchronized cells inhibits cell cycle reentry and the phosphorylation of the retinoblastoma protein (Rb), whereas in exponentially growing cells, HIN-1 induces apoptosis without apparent cell cycle arrest and effect on Rb phosphorylation. Investigation of multiple signaling pathways revealed that mitogen-induced phosphorylation and activation of AKT are inhibited in HIN-1-expressing cells. In addition, expression of constitutively activate AKT abrogates HIN-1-mediated growth arrest. Taken together, these studies provide further evidence that HIN-1 possesses tumor suppressor functions, and that these activities may be mediated through the AKT signaling pathway. PMID- 16266986 TI - Asbestos-induced lung inflammation and epithelial cell proliferation are altered in myeloperoxidase-null mice. AB - Asbestos fibers are carcinogens causing oxidative stress and inflammation, but the sources and ramifications of oxidant production by asbestos are poorly understood. Here, we show that inhaled chrysotile asbestos fibers cause increased myeloperoxidase activity in bronchoalveolar lavage fluids (BALF) and myeloperoxidase immunoreactivity in epithelial cells lining distal bronchioles and alveolar ducts, sites of initial lung deposition of asbestos fibers. In comparison with sham mice, asbestos-exposed myeloperoxidase-null (MPO-/-) and normal (MPO+/+) mice exhibited comparable increases in polymorphonuclear leukocytes, predominately neutrophils, in BALF after 9 days of asbestos inhalation. Differential cell counts on BALF revealed decreased proportions of macrophages and increased lymphocytes in all mice exposed to asbestos, but numbers were decreased overall in asbestos-exposed myeloperoxidase-null versus normal mice. Asbestos-associated lung inflammation in myeloperoxidase-null mice was reduced (P < or = 0.05) in comparison with normal asbestos-exposed mice at 9 days. Decreased lung inflammation in asbestos-exposed myeloperoxidase-null mice at 9 days was accompanied by increases (P < or = 0.05) in Ki-67- and cyclin D1 positive immunoreactive cells, markers of cell cycle reentry, in the distal bronchiolar epithelium. Asbestos-induced epithelial cell proliferation in myeloperoxidase-null mice at 30 days was comparable to that found at 9 days. In contrast, inflammation and epithelial cell proliferation in asbestos-exposed normal mice increased over time. These results support the hypothesis that myeloperoxidase status modulates early asbestos-induced oxidative stress, epithelial cell proliferation, and inflammation. PMID- 16266987 TI - Unexpected roles for pRb in mouse skin carcinogenesis. AB - The mouse skin carcinogenesis represents one of the best models for the understanding of malignant transformation, including the multistage nature of tumor development. The retinoblastoma gene product (pRb) plays a critical role in cell cycle regulation, differentiation, and inhibition of oncogenic transformation. In epidermis, Rb-/- deletion leads to proliferation and differentiation defects. Numerous evidences showed the involvement of the retinoblastoma pathway in this model. However, the actual role of pRb is still unknown. To study the possible involvement of pRb in keratinocyte malignant transformation, we have carried out two-stage chemical skin carcinogenesis on Rb(F19/F19) (thereafter Rb+/+) and Rb(F19/F19);K14Cre (thereafter Rb-/-) animals. Unexpectedly, we found that Rb-/- mice developed fewer and smaller papillomas than the Rb+/+ counterparts. Moreover, the small size of the pRb-deficient tumors is associated with an increase in the apoptotic index. Despite this, pRb deficient tumors display an increased conversion rate to squamous cell carcinomas. Biochemical analyses revealed that these characteristics correlate with the differential expression and activity of different pathways, including E2F/p19arf/p53, PTEN/Akt, c-jun NH2-terminal kinase/p38, and nuclear factor kappaB. Collectively, our findings show unexpected and hitherto nondescribed roles of pRb during the process of epidermal carcinogenesis. PMID- 16266988 TI - Identification of an aberrantly spliced form of HDMX in human tumors: a new mechanism for HDM2 stabilization. AB - The HDMX protein is closely related to HDM2 with which it shares different structural domains, particularly the p53 binding domain and the ring finger domain, where the two HDM proteins interact. Several oncogenic forms derived from splicing of HDM2 have been described in cancer. This work aimed at investigating whether analogous forms of HDMX exist in human tumors. Here, we report the characterization of an aberrantly spliced form of HDMX, HDMX211, isolated from the thyroid tumor cell line, ARO. HDMX211 binds and stabilizes the HDM2 protein. Although it lacks the p53 binding domain, HDMX211 also stabilizes p53 by counteracting its degradation by HDM2. However, the resulting p53 is transcriptionally inactive and increasingly associated to its inhibitor HDM2. Expression of HDMX211 strongly enhances the colony-forming ability of human cells in the presence or absence of wild-type p53. Conversely, depletion of HDMX211 by small interfering RNA significantly reduces the growth of ARO cells and increases their sensitivity to chemotherapy. Screening of lung cancer biopsies shows the presence of HDMX211 in samples that overexpress HDM2 protein, supporting a pathologic role for this new protein. This is the first evidence of a variant form of HDMX that has oncogenic potential independently of p53. HDMX211 reveals a new mechanism for overexpression of the oncoprotein HDM2. Most interestingly, it outlines a possible molecular explanation for a yet unclarified tumor phenotype, characterized by simultaneous overexpression of HDM2 and wild-type p53. PMID- 16266989 TI - Copy number aberrations in mouse breast tumors reveal loci and genes important in tumorigenic receptor tyrosine kinase signaling. AB - Receptor tyrosine kinase (RTK) signaling plays a key role in the development of breast cancer. Defining the genes and pathways in the RTK signaling network that are important regulators of tumorigenesis in vivo will unveil potential candidates for targeted therapeutics. To this end, we used microarray comparative genomic hybridization to identify and compare copy number aberrations in five mouse models of breast cancer induced by wild-type and mutated forms of oncogenic ErbB2 or the polyomavirus middle T antigen (PyMT). We observed distinct genomic alterations among the various models, including recurrent chromosome 11 amplifications and chromosome 4 deletions, syntenic with human 17q21-25 and 1p35 36, respectively. Expression of oncogenic Erbb2 (NeuNT) under control of the endogenous Erbb2 promoter results in frequent (85%) amplification at the Erbb2 locus with striking structural similarity to the human amplicon, resulting in overexpression of at least two of the genes, Erbb2 and Grb7. Chromosome 11 amplicons distal to Erbb2 arise in a model (DB) overexpressing a mutant variant of PyMT (Y315/322F) unable to activate phosphatidylinositol 3-kinase. These amplicons are not observed in DB hyperplasias or in tumors overexpressing wild type PyMT and result in overexpression of Grb2 and Itgb4. Distal chromosome 4 deletions occur in a significantly higher proportion of Erbb2 than PyMT tumors and encompass 14-3-3sigma (Stratifin), which is expressed at low or undetectable levels in the majority of NeuNT tumors. Our studies highlight loci and genes important in the regulation of tumorigenic RTK signaling in mammary epithelial cells in vivo. PMID- 16266991 TI - Loss of heterozygosity and somatic mutations of the glucocorticoid receptor gene are rarely found at relapse in pediatric acute lymphoblastic leukemia but may occur in a subpopulation early in the disease course. AB - Glucocorticoids are pivotal in the treatment of children with acute lymphoblastic leukemia (ALL) and have significant antileukemic effects in the majority of children. However, clinical resistance is a significant problem. Although cell line models implicate somatic mutations and loss of heterozygosity (LOH) of the glucocorticoid receptor (GR) gene as a mechanism of in vitro glucocorticoid resistance, the relevance of this mechanism as a cause of clinical resistance in children with ALL is not known. Mutational screening of all coding exons of the GR gene and LOH analyses were done in a large cohort of relapsed ALL. We show that somatic mutations and LOH of the GR rarely contribute to relapsed disease in children with ALL. However, we report the second case of ALL with a somatic mutation of the GR involving a 29-bp deletion in exon 8 and resulting in a truncated protein with loss of part of the ligand-binding domain. There was no evidence of a remaining wild-type allele. Allele-specific PCR detected the mutated clone at day 28 after presentation, which persisted at a low level throughout the disease course before relapse several years later. We hypothesize that the mutated allele present in a leukemic subclone at initial diagnosis was selected for during remission induction with glucocorticoids and contributed to the emergence of a glucocorticoid-resistant cell population. PMID- 16266992 TI - Mutations of C-RAF are rare in human cancer because C-RAF has a low basal kinase activity compared with B-RAF. AB - The protein kinase B-RAF is mutated in approximately 8% of human cancers. Here we show that presumptive mutants of the closely related kinase, C-RAF, were detected in only 4 of 545 (0.7%) cancer cell lines. The activity of two of the mutated proteins is not significantly different from that of wild-type C-RAF and these variants may represent rare human polymorphisms. The basal and B-RAF-stimulated kinase activities of a third variant are unaltered but its activation by RAS is significantly reduced, suggesting that it may act in a dominant-negative manner to modulate pathway signaling. The fourth variant has elevated basal kinase activity and is hypersensitive to activation by RAS but does not transform mammalian cells. Furthermore, when we introduce the equivalent of the most common cancer mutation in B-RAF (V600E) into C-RAF, it only has a weak effect on kinase activity and does not convert C-RAF into an oncogene. This lack of activation occurs because C-RAF lacks a constitutive charge within a motif in the kinase domain called the N-region. This fundamental difference in RAF isoform regulation explains why B-RAF is frequently mutated in cancer whereas C-RAF mutations are rare. PMID- 16266990 TI - Proteolysis of CCN1 by plasmin: functional implications. AB - Plasmin is shown to play a crucial role in many pathophysiologic processes primarily through its ability to degrade extracellular matrix (ECM) and/or mobilizing growth factors that are sequestered in the ECM. Cysteine-rich 61 (CCN1) is a matricellular protein of which expression is up-regulated in cancer and various vascular diseases. The present study was undertaken to investigate whether plasmin liberates CCN1 from the ECM and whether the released growth factor modulates endothelial cell migration. Treatment of breast carcinoma cells (MDA-MB-231) with plasmin released a truncated form of CCN1 (28 kDa) into the overlying medium. Experiments with recombinant CCN1 confirmed that plasmin effectively cleaves CCN1. Thrombin and other clotting/fibrinolytic proteases are ineffective in cleaving CCN1. Further studies revealed that the conditioned medium of plasmin-treated carcinoma cells supports endothelial cell migration and that antibodies specific to CCN1 blocked this enhancing effect. These data were the first to show that plasmin can liberate a pluripotent matrix signaling protein, CCN1, from the ECM. Because both CCN1 and the components of the plasmin generation system are present in tumor cells and a variety of other cells, the proteolysis of CCN1 by plasmin may play a role in many pathophysiologic processes, including tumor cell-mediated angiogenesis. PMID- 16266993 TI - Cyclin E overexpression obstructs infiltrative behavior in breast cancer: a novel role reflected in the growth pattern of medullary breast cancers. AB - Cell cycle deregulation is a prerequisite in tumor development and overexpression of cyclin E, a major G1-S regulator, is often observed in breast cancer and is further linked to poor prognosis. By overexpressing cyclin E in a retinoblastoma inactivated breast cancer cell line, we induced significant alterations in the expression of genes associated with proliferation and cell adhesion. Rearrangements of the actin cytoskeleton in addition to increased adhesive properties, decreased motility, and invasive potential in functional assays, indicated an overall abrogated mobility. Consistent in vivo findings were obtained upon investigation of 985 primary breast cancers, where cyclin E-high tumors predominantly (67%) displayed a low infiltrative, pushing growth pattern. Furthermore, medullary breast cancers, a subtype defined by its pushing, delimited growth, exhibited a remarkable frequency of cyclin E deregulation (87%) compared with other histologic subtypes (5-20%). Taken together, our results suggest the novel role of cyclin E in modeling infiltrative behavior. The consequences of cyclin E overexpression in breast cancer seems to be multiple, including effects on proliferation as well as growth patterns, a scenario that is indeed observed in the archetype of cyclin E-overexpressing medullary breast cancers. PMID- 16266994 TI - Induction of early post-transplant graft-versus-leukemia effects using intentionally mismatched donor lymphocytes and elimination of alloantigen-primed donor lymphocytes for prevention of graft-versus-host disease. AB - Graft-versus-leukemia (GVL) effects can be induced in tolerant mixed chimeras prepared with nonmyeloablative conditioning. GVL effects can be amplified by post grafting donor lymphocyte infusion (DLI). Unfortunately, DLI is frequently associated with graft-versus-host disease (GVHD). We investigated the feasibility of induction of potent GVL effects by DLI using intentionally mismatched lymphocytes followed by elimination of alloreactive donor T cells by cyclophosphamide for prevention of lethal GVHD following induction of very short yet most potent GVL effects. Mice inoculated with B-cell leukemia (BCL1) and mismatched donor lymphocytes were treated 2 weeks later with low-dose or high dose cyclophosphamide. All mice receiving cyclophosphamide 2 weeks after DLI survived GVHD, and no residual disease was detected by PCR; all control mice receiving DLI alone died of GVHD. Analysis of host (female) and donor (male) DNA showed that cyclophosphamide treatment eradicated most alloreactive donor cells, yet mixed chimerism was converted to full donor chimerism following transient self-limited GVHD. Our working hypothesis suggests that short-term yet effective and safe adoptive immunotherapy of leukemia may be accomplished early post transplantation using alloreactive donor lymphocytes, with prevention of GVHD by elimination of GVL effector cells. PMID- 16266995 TI - Pericytes and endothelial precursor cells: cellular interactions and contributions to malignancy. AB - Tumor vasculature is irregular, abnormal, and essential for tumor growth. Pericytes and endothelial precursor cells (EPC) contribute to the formation of blood vessels under angiogenic conditions. As primary cells in culture, pericytes and EPC share many properties such as tube/network formation and response to kinase inhibitors selective for angiogenic pathways. Expression of cell surface proteins including platelet-derived growth factor receptor, vascular cell adhesion molecule, intercellular adhesion molecule, CD105, desmin, and neural growth proteoglycan 2 was similar between pericytes and EPC, whereas expression of P1H12 and lymphocyte function-associated antigen-1 clearly differentiates the cell types. Further distinction was observed in the molecular profiles for expression of angiogenic genes. Pericytes or EPC enhanced the invasion of MDA-MB 231 breast cancer cells in a coculture assay system. The s.c. coinjection of live pericytes or EPC along with MDA-MB-231 cells resulted in an increased rate of tumor growth compared with coinjection of irradiated pericytes or EPC. Microvessel density analysis indicated there was no difference in MDA-MB-231 tumors with or without EPC or pericytes. However, immunohistochemical staining of vasculature suggested that EPC and pericytes may stabilize or normalize vasculature rather than initiate vasculogenesis. In addition, tumors arising from the coinjection of EPC and cancer cells were more likely to develop lymphatic vessels. These results support the notion that pericytes and EPC contribute to malignancy and that these cell types can be useful as cell-based models for tumor vascular development and selection of agents that may provide therapeutic benefit. PMID- 16266996 TI - Maternal embryonic leucine zipper kinase/murine protein serine-threonine kinase 38 is a promising therapeutic target for multiple cancers. AB - To identify genes that could serve as targets for novel cancer therapeutics, we used a bioinformatic analysis of microarray data comparing gene expression between normal and tumor-derived primary human tissues. From this approach, we have found that maternal embryonic leucine zipper kinase (Melk), a member of the AMP serine/threonine kinase family, exhibits multiple features consistent with the potential utility of this gene as an anticancer target. An oligonucleotide microarray analysis of multiple human tumor samples and cell lines suggests that Melk expression is frequently elevated in cancer relative to normal tissues, a pattern confirmed by quantitative reverse transcription-PCR and Western blotting of selected primary tumor samples. In situ hybridization localized Melk expression to malignant epithelial cells in 96%, 23%, and 13% of colorectal, lung, and ovarian tissue tumor samples, respectively. Expression of this gene is also elevated in spontaneous tumors derived from the ApcMin and Apc1638N murine models of intestinal tumorigenesis. To begin addressing whether Melk is relevant for tumorigenesis, RNA interference-mediated silencing within human and murine tumor cell lines was done. We show that Melk knockdown decreases proliferation and anchorage-independent growth in vitro as well as tumor growth in a xenograft model. Together, these results suggest that Melk may provide a growth advantage for neoplastic cells and, therefore, inactivation may be therapeutically beneficial. PMID- 16266997 TI - Expression of Frzb/secreted Frizzled-related protein 3, a secreted Wnt antagonist, in human androgen-independent prostate cancer PC-3 cells suppresses tumor growth and cellular invasiveness. AB - The ability of Frzb/secreted Frizzled-related protein 3 (sFRP3) to inhibit Wnt signaling and the localization of Frzb/sFRP3 on chromosome 2q to a region frequently deleted in cancers have led some investigators to hypothesize that Frzb/sFRP3 is a tumor suppressor gene. Here, we examined the biological effects of Frzb/sFRP3 on an androgen-independent prostate cancer cell model. We showed that expression of Frzb/sFRP3 in PC-3 cells resulted in decreased colony formation in soft agar and a dramatic inhibition of tumor growth in a xenograft mouse model. When cellular morphology was examined, PC-3 cells expressing Frzb/sFRP3 exhibited an increase in cell-cell contact formation accompanied by a pronounced induction of epithelial markers E-cadherin and keratin-8 and down regulation of mesenchymal markers N-cadherin, fibronectin, and vimentin. This phenomenon suggested a reversal of epithelial-to-mesenchymal transition and a less invasive phenotype. Indeed, further in vitro studies with a Matrigel assay showed that Frzb/sFRP3 decreased the invasive capacity of PC-3 cells. These changes in the biology of PC-3 cells are associated with a decrease in the expression and activities of both matrix metalloproteinase (MMP)-2 and MMP-9 as well as decreases in AKT activation, cytosolic beta-catenin levels, T-cell factor transcription activity, and expression of Slug and Twist. In addition, transfection of PC-3 with a dominant-negative low-density lipoprotein receptor related protein 5 (DN-LRP5) coreceptor showed similar biological effects as Frzb/sFRP3 transfection. Together, these data suggest that Frzb/sFRP3 and DN-LRP5 exhibit antitumor activity through the reversal of epithelial-to-mesenchymal transition and inhibition of MMP activities in a subset of prostate cancer. PMID- 16266998 TI - Suberoylanilide hydroxamic acid enhances gap junctional intercellular communication via acetylation of histone containing connexin 43 gene locus. AB - A histone deacetylase (HDAC) inhibitor, suberoylanilide hydroxamic acid (SAHA), induces apoptosis in neoplastic cells, but its effect on gap junctional intercellular communication in relation to apoptosis was unclear. Therefore, we carried out a comparative study of the effects of two HDAC inhibitors, SAHA and trichostatin-A, on gap junctional intercellular communication in nonmalignant human peritoneal mesothelial cells (HPMC) and tumorigenic ras oncogene transformed rat liver epithelial cells (WB-ras) that showed a significantly lower level of gap junctional intercellular communication than did HPMC. Gap junctional intercellular communication was assessed by recovery rate of fluorescence recovery after photobleaching. Treatment of HPMC with SAHA at nanomolar concentrations caused a dose-dependent increase of recovery rate without inducing apoptosis. This effect was accompanied by enhanced connexin 43 (Cx43) mRNA and protein expression and increased presence of Cx43 protein on cell membrane. Trichostatin-A induced apoptosis in HPMC but was less potent than SAHA in enhancing the recovery rate. In contrast, treatment of WB-ras cells with SAHA or trichostatin-A induced apoptosis at low concentrations, in spite of smaller increases in recovery rate, Cx43 mRNA, and protein than in HPMC. Chromatin immunoprecipitation analysis revealed that SAHA enhanced acetylated histones H3 and H4 in the chromatin fragments associated with Cx43 gene in HPMC. These results indicate that SAHA at low concentrations selectively up-regulates Cx43 expression in normal human cells without induction of apoptosis, as a result of histone acetylation in selective chromatin fragments, in contrast to the apoptotic effect observed in tumorigenic WB-ras cells. These results support a cancer therapeutic and preventive role for specific HDAC inhibitors. PMID- 16266999 TI - Progestins initiate a luminal to myoepithelial switch in estrogen-dependent human breast tumors without altering growth. AB - Although long-term clinical use of progestins is associated with an increased incidence of breast cancers, their role in established cancers is unclear. Estrogens are considered to be the main mitogens in the majority of breast cancers. Whether progesterone affects proliferation and/or differentiation is under debate. To assess the role of progesterone in established breast cancers, we used T47D human breast cancer cells that are estrogen receptor (ER) positive and either progesterone receptor (PR) negative or positive for PRA, PRB, or both. These cells were grown as strictly estrogen-dependent solid tumors in ovariectomized female nude mice. Progesterone or medroxyprogesterone acetate (MPA) alone did not support tumor growth, nor did progesterone or MPA given simultaneously with estrogen significantly alter estrogen-dependent tumor growth. However, treatment of mice bearing ER+PR+ but not ER+PR- tumors with either progesterone or MPA increased expression of the myoepithelial cytokeratins (CK) 5 and 6 in a subpopulation of tumor cells. These CK5+/CK6+ cells had decreased expression of luminal epithelial CK8, CK18, and CK19. We conclude that progestins exert differentiative effects on tumors characterized by transition of a cell subpopulation from luminal to myoepithelial. This may not be beneficial, however, because such a phenotype is associated with poor prognosis. PMID- 16267000 TI - Tumor-secreted vascular endothelial growth factor-C is necessary for prostate cancer lymphangiogenesis, but lymphangiogenesis is unnecessary for lymph node metastasis. AB - Dissemination to draining lymph nodes is a frequent first step in prostate cancer metastasis. Although tumors metastasize to lymph nodes via the lymphatics, the importance of lymphangiogenesis in mediating the process remains controversial. Here, we inhibit intratumoral lymphangiogenesis in s.c. and surgical orthotopic implantation mouse models of human prostate cancer using several strategies. Stable expression of small interfering RNAs (siRNA) targeted against human vascular endothelial growth factor-C (VEGF-C) in PC-3 cells reduced intratumoral lymphatics by 99% in s.c. tumors, indicating that tumor-secreted VEGF-C is necessary for lymphangiogenesis. Expression of siRNAs against human VEGF-A somewhat reduced tumor lymphangiogenesis. Secretion of a soluble VEGF receptor 3/Flt4 fusion protein by PC-3 cells reduced intratumoral lymphatics by 100% in s.c. tumors. Combination of soluble Flt4 and VEGF-C siRNA yielded >92% reduction of intratumoral lymphatics in orthotopic prostate tumors. However, metastasis to lymph nodes was not significantly affected regardless of intratumoral lymphatic vessel density. The abundance of marginal lymphatics at the tumor-stromal interface was unchanged in orthotopic tumors whose intratumoral lymphatics were inhibited, suggesting that these marginal vessels could be sufficient for lymph node metastasis. Hematogenous metastasis (blood tumor burden, lung metastasis) correlated with degree of lymph node invasion. We also analyzed the lymphatics in spontaneous transgenic adenocarcinomas of the mouse prostate which metastasize to lymph nodes. Progression from well-differentiated prostate intraepithelial neoplasia to metastatic, undifferentiated adenocarcinoma was accompanied by loss of lymphatics. These results suggest that tumor-secreted VEGF-C and, to a lesser extent, VEGF-A, are important for inducing prostate cancer intratumoral lymphangiogenesis but are unnecessary for lymph node metastasis. PMID- 16267001 TI - A role for polymerase eta in the cellular tolerance to cisplatin-induced damage. AB - Mutation of the POLH gene encoding DNA polymerase eta (pol eta) causes the UV sensitivity syndrome xeroderma pigmentosum-variant (XP-V) which is linked to the ability of pol eta to accurately bypass UV-induced cyclobutane pyrimidine dimers during a process termed translesion synthesis. Pol eta can also bypass other DNA damage adducts in vitro, including cisplatin-induced intrastrand adducts, although the physiological relevance of this is unknown. Here, we show that independent XP-V cell lines are dramatically more sensitive to cisplatin than the same cells complemented with functional pol eta. Similar results were obtained with the chemotherapeutic agents, carboplatin and oxaliplatin, thus revealing a general requirement for pol eta expression in providing tolerance to these platinum-based drugs. The level of sensitization observed was comparable to that of XP-A cells deficient in nucleotide excision repair, a recognized and important mechanism for repair of cisplatin adducts. However, unlike in XP-A cells, the absence of pol eta expression resulted in a reduced ability to overcome cisplatin induced S phase arrest, suggesting that pol eta is involved in translesion synthesis past these replication-blocking adducts. Subcellular localization studies also highlighted an accumulation of nuclei with pol eta foci that correlated with the formation of monoubiquitinated proliferating cell nuclear antigen following treatment with cisplatin, reminiscent of the response to UV irradiation and further indicating a role for pol eta in dealing with cisplatin induced damage. Together, these data show that pol eta represents an important determinant of cellular responses to cisplatin, which could have implications for acquired or intrinsic resistance to this key chemotherapeutic agent. PMID- 16267002 TI - Tocopherol-associated protein suppresses prostate cancer cell growth by inhibition of the phosphoinositide 3-kinase pathway. AB - Epidemiologic studies suggested that vitamin E has a protective effect against prostate cancer. We showed here that tocopherol-associated protein (TAP), a vitamin E-binding protein, promoted vitamin E uptake and facilitated vitamin E antiproliferation effect in prostate cancer cells. Interestingly, without vitamin E treatment, overexpression of TAP in prostate cancer cells significantly suppressed cell growth; knockdown of endogenous TAP by TAP small interfering RNA (siRNA) in nonmalignant prostate HPr-1 cells increased cell growth. Further mechanism dissection studies suggested that the tumor suppressor function of TAP was via down-regulation of phosphoinositide 3-kinase (PI3K)/Akt signaling, but not by modulating cell cycle arrest or androgen receptor signaling. Immunoprecipitation results indicated that TAP inhibited the interaction of PI3K subunits, p110 with p85, and subsequently reduced Akt activity. Constitutively active Akt could negate the TAP-suppressive activity on prostate cancer cell growth. Moreover, stable transfection of TAP in LNCaP cells suppressed LNCaP tumor incidence and growth rate in nude mice. Furthermore, TAP mRNA and protein expression levels were significantly down-regulated in human prostate cancer tissue samples compared with benign prostate tissues as measured by reverse transcription-PCR, in situ hybridization, and immunohistochemistry. Together, our data suggest that TAP not only mediates vitamin E absorption to facilitate vitamin E antiproliferation effect in prostate cancer cells, but also functions like a tumor suppressor gene to control cancer cell viability through a non vitamin E manner. Therefore, TAP may represent a new prognostic marker for prostate cancer progression. PMID- 16267003 TI - Addressing the "Fas counterattack" controversy: blocking fas ligand expression suppresses tumor immune evasion of colon cancer in vivo. AB - Fas ligand (FasL/CD95L) is a transmembrane protein belonging to the tumor necrosis factor superfamily that can trigger apoptotic cell death following ligation to its receptor, Fas (CD95/APO-1). Expression of FasL may help to maintain tumor cells in a state of immune privilege by inducing apoptosis of antitumor immune effector cells-the "Fas counterattack." However, the ability of FasL to mediate tumor immune privilege is controversial due to studies that indicate FasL has both pro- and anti-inflammatory activities. To resolve this controversy and functionally define the role of FasL in tumor immune evasion, we investigated if suppression of endogenously expressed FasL in colon tumor cells resulted in reduced tumor development and improved antitumor immune challenge in vivo. Specifically, FasL expression in CMT93 colon carcinoma cells was down regulated following stable transfection with a plasmid encoding antisense FasL cDNA. Down-regulation of FasL expression had no effect on tumor growth in vitro but significantly reduced tumor development in syngeneic immunocompetent mice in vivo. Tumor size was also significantly decreased. Reduced FasL expression by tumor cells led to increased lymphocyte infiltration. The overall level of neutrophils present in all of the tumors examined was low, with no difference between the tumors, irrespective of FasL expression. Thus, down-regulation of FasL expression by colon tumor cells results in an improved antitumor immune challenge in vivo, providing functional evidence in favor of the "Fas counterattack" as a mechanism of tumor immune evasion. PMID- 16267004 TI - A network of genetic events sufficient to convert normal human cells to a tumorigenic state. AB - Although great progress has been made at identifying and characterizing individual genes involved in cancer, less is known about how the combination of such genes collaborate to form tumors in humans. To this end, we sought to genetically recreate tumorigenesis in normal human cells using genes altered in human cancer. We now show that expression of mammalian proteins that inactivate the tumor suppressors Rb and p53 in conjunction with the oncoproteins Ras and Myc and the telomerase subunit hTERT is sufficient to drive a number of normal human somatic cells to a tumorigenic fate. This provides a blueprint of the events that lead to human cancer, allowing different cancers to be genetically modeled from normal human cells. PMID- 16267005 TI - High correlation of whole-body red fluorescent protein imaging and magnetic resonance imaging on an orthotopic model of pancreatic cancer. AB - We have developed genetically fluorescent orthotopic models of human pancreatic cancer. In these models, noninvasive fluorescent protein imaging (FPI) of internal primary tumors and metastatic deposits has been carried out. Whole-body tumor images are easily and inexpensively obtained using FPI, permitting both detection and quantification of tumor load. In this study, we simultaneously compared single mice with a highly fluorescent, red fluorescent protein expressing orthotopic pancreatic cancer xenografts with both FPI and high resolution magnetic resonance imaging (MRI). Images were acquired at multiple time points after tumor implantation in the pancreas. Indwelling pancreatic primary tumors and metastatic foci were detected by both FPI and MRI. Moreover, a strong correlation existed between images taken with these two technologies. FPI permitted rapid, high-throughput imaging without the need for either anesthesia or contrast agents. Both FPI and MRI enabled accurate imaging of tumor growth and metastasis, although MRI enabled tissue structure to be visualized as well. FPI has high resolution and is exceedingly rapid with instant image capture. We suggest a complimentary role for these two imaging modalities. PMID- 16267006 TI - DNA damage disrupts the p14ARF-B23(nucleophosmin) interaction and triggers a transient subnuclear redistribution of p14ARF. AB - The p14 alternate reading frame (ARF) tumor suppressor plays a central role in cancer by binding to mdm2 (Hdm2 in humans) and enhancing p53-mediated apoptosis following DNA damage and oncogene activation. It is unclear, however, how ARF initiates its involvement in the p53/mdm2 pathway, as p53 and mdm2 are located in the nucleoplasm, whereas ARF is largely nucleolar in tumor cells. We have used immunofluorescence and coimmunoprecipitation to examine how the subnuclear distribution and protein-protein interactions of ARF change immediately after DNA damage and over the time course of the DNA damage response in human tumor cells. We find that DNA damage disrupts the interaction of ARF with the nucleolar protein B23(nucleophosmin) and promotes a transient p53-independent translocation of ARF to the nucleoplasm, resulting in a masking of the ARF NH2 terminus that correlates with the appearance of ARF-Hdm2 complexes. The translocation also results in an unmasking of the ARF COOH terminus, suggesting that redistribution disrupts a nucleolar interaction of ARF involving this region. By 24 hours after irradiation, DNA repair has ceased and the pretreatment immunofluorescence patterns and complexes of ARF have been restored. Although the redistribution of ARF is independent of p53 and likely to be regulated by interactions other than Hdm2, ARF does not promote UV sensitization unless p53 is expressed. The results implicate the nucleolus and nucleolar interactions of the ARF, including potentially novel interactions involving its COOH terminus as sites for early DNA damage and stress-mediated cellular events. PMID- 16267007 TI - Cerebrospinal fluid proteomic analysis reveals dysregulation of methionine aminopeptidase-2 expression in human and mouse neurofibromatosis 1-associated glioma. AB - Individuals affected with the neurofibromatosis 1 (NF1) tumor predisposition syndrome are prone to the development of multiple nervous system tumors, including optic pathway gliomas (OPG). The NF1 tumor suppressor gene product, neurofibromin, functions as a Ras GTPase-activating protein, and has been proposed to regulate cell growth by inhibiting Ras activity. Recent studies from our laboratory have shown that neurofibromin also regulates the mammalian target of rapamycin activity in a Ras-dependent fashion, and that the rapamycin-mediated mammalian target of rapamycin inhibition ameliorates the Nf1-/- astrocyte growth advantage. Moreover, Nf1-deficient astrocytes exhibit increased protein translation. As part of a larger effort to identify protein markers for NF1 associated astrocytomas that could be exploited for therapeutic drug design, we did an objective proteomic analysis of the cerebrospinal fluid from genetically engineered Nf1 mice with optic glioma. One of the proteins found to be increased in the cerebrospinal fluid of OPG-bearing mice was the eukaryotic initiation factor-2alpha binding protein, methionine aminopeptidase 2 (MetAP2). In this study, we show that Nf1 mouse OPGs and NF1-associated human astrocytic tumors, but not sporadic pilocytic or other low-grade astrocytomas, specifically expressed high levels of MetAP2. In addition, we show that Nf1-deficient astrocytes overexpress MetAP2 in vitro and in vivo, and that treatment with the MetAP2 inhibitor fumagillin significantly reduces Nf1-/- astrocyte proliferation in vitro. These observations suggest that MetAP2 is regulated by neurofibromin, and that MetAP2 inhibitors could be potentially employed to treat NF1-associated tumor proliferation. PMID- 16267009 TI - The intrinsic mitochondrial membrane potential of colonic carcinoma cells is linked to the probability of tumor progression. AB - We subcloned cell lines from SW620 cells establishing that, despite the dynamic nature of the mitochondrial membrane potential (Deltapsim), there are significant and stable differences in the intrinsic Deltapsim among cells within an in vitro population of human colonic carcinoma cells. Whereas more dramatic differences in Deltapsim would likely perturb essential mitochondrial functions, the differences in Deltapsim of the subclones did not affect steady-state reactive oxygen species levels, electron transport activity, or cellular viability and growth rates. However, the differences in intrinsic Deltapsim had a significant effect on the tumorigenic behavior of the cells. Subcloned cell lines with higher Deltapsim were more likely to exhibit elevated steady-state levels of vascular endothelial growth factor and matrix metalloproteinase 7, and increased invasive behavior (properties associated with tumor progression), than cells with lower intrinsic Deltapsim, whereas cells with lower Deltapsim were more likely to respond to the chemopreventive activities of butyrate, including Deltapsim dissipation, growth arrest, and apoptosis, than cells with higher Deltapsim. Therefore, these data establish that the probability for tumor development and progression is linked to stable differences in the intrinsic Deltapsim of colonic epithelial cells. PMID- 16267008 TI - Focal adhesion kinase promotes the aggressive melanoma phenotype. AB - Malignant melanoma continues to remain a significant health threat, with death often occurring as a result of metastasis. The metastatic phenotype typically is characterized by augmented tumor cell invasion and migration in addition to tumor cell plasticity as shown by vasculogenic mimicry. Therefore, understanding the molecular mechanisms that promote an aggressive phenotype is essential to predicting the likelihood of metastasis at a stage when intervention may be possible. This study focuses on the role of focal adhesion kinase (FAK), a cytoplasmic tyrosine kinase important for many cellular processes, including cell survival, invasion, and migration. We found FAK to be phosphorylated on its key tyrosine residues, Tyr397 and Tyr576, in only aggressive uveal and cutaneous melanoma cells, which correlates with their increased invasion, migration, and vasculogenic mimicry plasticity. Additionally, we confirmed the presence of FAK phosphorylated on Tyr397 and Tyr576 in both cutaneous and uveal melanoma tumors in situ. Examination of a functional role for FAK in aggressive melanoma revealed that disruption of FAK-mediated signal transduction pathways, through the expression of FAK-related nonkinase (FRNK), results in a decrease in melanoma cell invasion, migration, and inhibition of vasculogenic mimicry. Moreover, we found that FRNK expression resulted in a down-regulation of Erk1/2 phosphorylation resulting in a decrease in urokinase activity. Collectively, these data suggest a new mechanism involved in promoting the aggressive melanoma phenotype through FAK-mediated signal transduction pathways, thus providing new insights into possible therapeutic intervention strategies. PMID- 16267010 TI - Proliferation of human neuroblastomas mediated by the epidermal growth factor receptor. AB - Neuroblastoma is a common solid tumor of childhood that is derived from the neural crest. Expression of epidermal growth factor (EGF) receptors (EGFRs) has been associated with enhanced cell growth and aggressive behavior in other tumors. Here, we examined the expression profile of EGFRs in neuroblastoma cell lines and primary tumors. We found that all 13 neuroblastoma cell lines examined expressed EGFR1 (HER1), most at readily detectable levels. Low levels of other human EGFR family receptors were also detected in almost all cell lines. All primary tumors examined expressed readily detectable levels of HER1 and HER3 and lower levels of HER2 and HER4. EGF had a significant effect on the proliferation of neuroblastoma cell lines in vitro. EGF treatment (100 ng/mL) of the cell lines SY5Y and NLF significantly increased cell number (P < 0.01). EGF stimulated more cells to enter S and G2-M phase, as suggested by flow cytometry, indicating that EGF increases cell number by increasing proliferation, with no appreciable change in apoptosis. EGF exposure resulted in receptor autophosphorylation and activation of both the mitogen-activated protein kinase (MAPK) and phosphoinositide 3-kinase (PI3K)/AKT pathways. Exposure to 0.5 micromol/L ZD1839, a HER1-specific inhibitor, caused a 40% to 50% reduction in the number of SY5Y and NLF cells grown in medium containing 10% fetal bovine serum (P < 0.01). Even at 0.01 micromol/L, ZD1839 inhibited autophosphorylation of HER1 by EGF. At 0.1 micromol/L, it also blocked phosphorylation of AKT, but not MAPK, in NLF cells. Additional studies showed that the PI3K/AKT-specific inhibitor LY294002 had a more profound effect than the MAPK-specific inhibitor U0126 in blocking EGF induced cell proliferation. This suggests that the PI3K/AKT pathway is the main signaling pathway responsible for the proliferation effects of EGF in neuroblastomas. Our results also indicate that ZD1839 is a potent inhibitor of neuroblastoma cell proliferation; therefore, it may be a useful, biologically based therapeutic agent for these tumors. PMID- 16267011 TI - Assessment of low linear energy transfer radiation-induced bystander mutagenesis in a three-dimensional culture model. AB - A three-dimensional cell culture model composed of human-hamster hybrid (A(L)) and Chinese hamster ovary (CHO) cells in multicellular clusters was used to investigate low linear energy transfer (LET) radiation-induced bystander genotoxicity. CHO cells were mixed with A(L) cells in a 1:5 ratio and briefly centrifuged to produce a spheroid of 4 x 10(6) cells. CHO cells were labeled with tritiated thymidine ([3H]dTTP) for 12 hours and subsequently incubated with A(L) cells for 24 hours at 11 degrees C. The short-range beta-particles emitted by [3H]dTTP result in self-irradiation of labeled CHO cells; thus, biological effects on neighboring A(L) cells can be attributed to the bystander response. Nonlabeled bystander A(L) cells were isolated from among labeled CHO cells by using a magnetic separation technique. Treatment of CHO cells with 100 microCi [3H]dTTP resulted in a 14-fold increase in bystander mutation incidence among neighboring A(L) cells compared with controls. Multiplex PCR analysis revealed the types of mutants to be significantly different from those of spontaneous origin. The free radical scavenger DMSO or the gap junction inhibitor Lindane within the clusters significantly reduced the mutation incidence. The use of A(L) cells that are dominant negative for connexin 43 and lack gap junction formation produced a complete attenuation of the bystander mutagenic response. These data provide evidence that low LET radiation can induce bystander mutagenesis in a three-dimensional model and that reactive oxygen species and intercellular communication may have a modulating role. The results of this study will address the relevant issues of actual target size and radiation quality and are likely to have a significant effect on our current understanding of radiation risk assessment. PMID- 16267012 TI - Rac3-mediated transformation requires multiple effector pathways. AB - Our initial characterization of Rac3, a close relative of the small GTPase Rac1, established its ability to promote membrane ruffling, transformation, and activation of c-jun transcriptional activity. The finding that Rac3 is transforming, and its similarity to Rac1, a protein that has a well-established connection to many processes important for cancer progression, prompted further investigation into Rac3 transformation. We used effector domain mutants (EDMs) to explore the relationship among Rac signaling, transformation, and effector usage. All Rac3 EDMs tested (N26D, F37L, Y40C, and N43D) retained the ability to promote membrane ruffling and focus formation. In contrast, only the N43D mutant promoted anchorage independence. This differs from Rac1, where both N26D and N43D mutants were impaired in both types of transformation. To learn more about the signaling pathways involved, we did luciferase reporter assays and glutathione S transferase pull-down assays for effector binding. We found evidence for a functional link between activation of phospholipase Cbeta2 by Rac3 and signaling to the serum response factor (SRF). Surprisingly, we also found that Rac3 binds poorly to the known Rac1 effectors mixed lineage kinases 2 and 3 (MLK2 and MLK3). Transcription of cyclin D1 was the only pathway that correlated with growth in soft agar. Our experiments show that activation of membrane ruffling and transcriptional activation of c-jun, SRF, or E2F are not sufficient to promote anchorage-independent growth mediated by Rac3. Instead, multiple effector pathways are required for Rac3 transformation, and these overlap partially but not completely with those used by Rac1. PMID- 16267013 TI - Up-regulation of CXCR4 expression in PC-3 cells by stromal-derived factor-1alpha (CXCL12) increases endothelial adhesion and transendothelial migration: role of MEK/ERK signaling pathway-dependent NF-kappaB activation. AB - The chemokine stromal-derived factor-1alpha (SDF-1alpha/CXCL-12) and its receptor, CXCR4, play a crucial role in adhesion and transendothelium migration (TEM) of prostate cancer cells. We tested the hypothesis that enhanced expression of CXCR4 in prostate cancer cells is dependent upon SDF-1alpha-mediated activation of nuclear factor-kappaB (NF-kappaB). SDF-1alpha increased the CXCR4 mRNA and protein expression in PC-3 cells but not in LNCaP cells. Similarly, SDF 1alpha enhanced the NF-kappaB-dependent transcriptional activity in PC-3 cells but not in LNCaP cells. SDF-1alpha increased PC-3 cell adhesion to the human umbilical vein endothelial cell monolayer and enhanced TEM, which was abrogated with anti-CXCR4 monoclonal antibody (mAb). Suppression of NF-kappaB activity in PC-3 cells by a mutant IkappaBalpha super-repressor adenoviral vector decreased the CXCR4 mRNA expression and inhibited adhesion and TEM. Transient overexpression of p65 subunit of NF-kappaB in PC-3 cells up-regulated CXCR4 receptor expression and increased the adhesion and TEM of these cells in response to SDF-1alpha gradient. Treatment of PC-3 cells with SDF-1alpha leads to nuclear translocation of NF-kappaB protein within 15 to 30 minutes, which correlated with IkappaBalpha phosphorylation. A p42/44 mitogen-activated protein kinase [MAPK, extracellular signal regulated kinase-1/2 (ERK-1/2)] biphasic activation pattern was observed in these cells at 15 minutes and 3 hours after SDF-1alpha treatment. Phosphorylation of IkappaB kinase alpha was observed within 30 minutes, which was blocked by PD98059 [MAPK kinase (MEK) inhibitor]. PD98059 cotreatment significantly inhibited SDF-1alpha-induced NF-kappaB reporter activity and CXCR4 receptor expression as shown by flow cytometry. These data suggest that SDF 1alpha-induced expression of CXCR4 in PC-3 cells is dependent on MEK/ERK signaling cascade and NF-kappaB activation. PMID- 16267014 TI - Focal adhesion kinase regulates syndecan-2-mediated tumorigenic activity of HT1080 fibrosarcoma cells. AB - Expression of syndecan-2, a transmembrane heparan sulfate proteoglycan, is crucial for the tumorigenic activity in colon carcinoma cells. However, despite the high-level expression of syndecan-2 in mesenchymal cells, few studies have addressed the function of syndecan-2 in sarcoma cells. In HT1080 fibrosarcoma cells, we found that syndecan-2 regulated migration, invasion into Matrigel, and anchorage-independent growth but not cell-extracellular matrix adhesion or proliferation, suggesting that syndecan-2 plays different functional roles in fibrosarcoma and colon carcinoma cells. Consistent with the increased cell migration/invasion of syndecan-2-overexpressing HT1080 cells, syndecan-2 overexpression increased phosphorylation and interaction of focal adhesion kinase (FAK) and phosphatidylinositol 3-kinase (PI3K), membrane localization of T lymphoma invasion and metastasis gene-1 (Tiam-1), and activation of Rac. Syndecan 2-mediated cell migration/invasion of HT1080 cells was diminished when (a) cells were cotransfected with nonphosphorylatable mutant FAK Y397F or with other FAK mutants lacking PI3K interactions, (b) cells were treated with a specific PI3K inhibitor, or (c) levels of Tiam-1 were knocked down with small interfering RNAs. Furthermore, expression of several FAK mutants inhibited syndecan-2-mediated enhancement of anchorage-independent growth in HT1080 cells. Taken together, these data suggest that syndecan-2 regulates the tumorigenic activities of HT1080 fibrosarcoma cells and that FAK is a key regulator of syndecan-2-mediated tumorigenic activities. PMID- 16267015 TI - The role of DOC-2/DAB2 in modulating androgen receptor-mediated cell growth via the nongenomic c-Src-mediated pathway in normal prostatic epithelium and cancer. AB - Prostate cancer is initially responsive to androgen ablation, but prostate cancer tumors invariably progress to an androgen-independent state that is ultimately lethal. The onset of the androgen-independent prostate cancer is often associated with up-regulation of the androgen receptor that can cause antagonists to exhibit agonistic activity, which could lead to the failure of androgen ablation therapy. We describe a unique protein-DOC-2/DAB2 (differentially expressed in ovarian cancer-2/disabled 2)-that antagonizes androgen receptor-mediated cell growth in prostate cancer cells via interaction with c-Src protein. This interaction causes inactivation of Erk and Akt proteins critical for proliferation and survival of prostate cancer cells. However, DOC-2/DAB2 does not change the capacity of androgen receptor to regulate the transcription of androgen-responsive reporter genes, indicating that DOC-2/DAB2 selectively inhibits androgen receptor-mediated cell growth in androgen-independent prostate cancer by disrupting the androgen receptor/c-Src complex. In normal prostatic epithelia, DOC-2/DAB2 protein levels are more abundant than androgen receptor protein levels and reduced endogenous DOC-2/DAB2 protein levels in these cells by DOC-2/DAB2 RNA interference result in enhancing androgen receptor-mediated cell growth. We conclude that DOC-2/DAB2 can modulate androgen receptor-mediated cell growth in both normal and malignant prostatic epithelial cells and the outcome of this study could evolve into a new therapeutic strategy of prostate cancer. PMID- 16267016 TI - Induction of hepatocyte growth factor/scatter factor by fibroblast clustering directly promotes tumor cell invasiveness. AB - For determining the malignant behavior of a tumor, paracrine interactions between stromal and cancer cells are crucial. We previously reported that fibroblast clustering induces cyclooxygenase-2 (COX-2), plasminogen activation, and programmed necrosis, all of which were significantly reduced by nonsteroidal anti inflammatory drugs (NSAID). We have now found that tumor cell-conditioned medium induces similar fibroblast clustering. Activation of the necrotic pathway in clustering fibroblasts, compared with control monolayer cultures, induced a massive >200-fold production of bioactive hepatocyte growth factor/scatter factor (HGF/SF), which made human carcinoma cells spread and invade a collagen lattice. This response occurred only if a functional, properly processed c-Met receptor was present, which was then rapidly phosphorylated. The invasion-promoting activity was inhibited by a neutralizing HGF/SF antibody. NSAIDs, if added early during fibroblast aggregation, inhibited HGF/SF production effectively but had no effect at later stages of cell aggregation. Our results thus provide the first evidence that aggravated progression of tumors with necrotic foci may involve paracrine reciprocal signaling leading to stromal activation by direct cell-cell contact (i.e., nemosis). PMID- 16267017 TI - Retinol inhibits the growth of all-trans-retinoic acid-sensitive and all-trans retinoic acid-resistant colon cancer cells through a retinoic acid receptor independent mechanism. AB - Retinol (vitamin A) is thought to exert its effects through the actions of its metabolite, all-trans-retinoic acid (ATRA), on gene transcription mediated by retinoic acid receptors (RAR) and retinoic acid response elements (RARE). However, retinoic acid resistance limits the chemotherapeutic potential of ATRA. We examined the ability of retinol to inhibit the growth of ATRA-sensitive (HCT 15) and ATRA-resistant (HCT-116, SW620, and WiDR) human colon cancer cell lines. Retinol inhibited cell growth in a dose-responsive manner. Retinol was not metabolized to ATRA or any bioactive retinoid in two of the cell lines examined. HCT-116 and WiDR cells converted a small amount of retinol to ATRA; however, this amount of ATRA was unable to inhibit cell growth. To show that retinol was not inducing RARE-mediated transcription, each cell line was transfected with pRARE chloramphenicol acetyltransferase (CAT) and treated with ATRA and retinol. Although treatment with ATRA increased CAT activity 5-fold in ATRA-sensitive cells, retinol treatment did not increase CAT activity in any cell line examined. To show that growth inhibition due to retinol was ATRA, RAR, and RARE independent, a pan-RAR antagonist was used to block RAR signaling. Retinol induced growth inhibition was not alleviated by the RAR antagonist in any cell line, but the antagonist alleviated ATRA-induced growth inhibition of HCT-15 cells. Retinol did not induce apoptosis, differentiation or necrosis, but affected cell cycle progression. Our data show that retinol acts through a novel, RAR-independent mechanism to inhibit colon cancer cell growth. PMID- 16267018 TI - Bombesin regulates cyclin D1 expression through the early growth response protein Egr-1 in prostate cancer cells. AB - Our previous studies indicate that the activation of mitogen-activated protein kinase (MAPK) pathway is involved in bombesin-induced cell proliferation in prostate cancer cells. Cyclin D1 is a critical regulator involved in cell cycle progression through the G1 phase into the S phase, thereby contributing to cell proliferation. Mostly, mitogen-stimulated expression of cyclin D1 is attributed to the extracellular signal-regulated kinase (ERK) activation. Here, we found that bombesin induced human cyclin D1 expression on both mRNA and protein levels in DU-145 prostate cancer cells. Mutational analyses showed that bombesin enhanced cyclin D1 transcription required the binding of nuclear proteins to the 143 to -105 region of the human cyclin D1 promoter, which contains binding sites for transcription factors Sp-1 and early growth response protein (Egr-1). Do novo protein synthesis was requisite for bombesin-induced cyclin D1 expression. Further studies showed Egr-1 was induced upon bombesin stimulation. The induction of Egr-1 expression and its binding to the cyclin D1 promoter were essential for bombesin-enhanced cyclin D1 transcription. Inhibition of MAPK pathway with either the MEK1 inhibitor PD98059 or a dominant-negative Ras mutant, RasN17, abolished bombesin-induced cyclin D1 activation. Taken together, bombesin-induced cyclin D1 expression in prostate cancer cells is mediated by Egr-1 activation and the interaction of Egr-1 with the Egr-1/Sp1 motif of the cyclin D1 promoter through the activation of MAPK pathway. These findings represent a novel mechanism of bombesin-dependent stimulation of mitogenesis by regulating directly the cell cycle in prostate cancer. PMID- 16267019 TI - Resveratrol inhibits myeloma cell growth, prevents osteoclast formation, and promotes osteoblast differentiation. AB - Multiple myeloma is characterized by the accumulation of clonal malignant plasma cells in the bone marrow, which stimulates bone destruction by osteoclasts and reduces bone formation by osteoblasts. In turn, the changed bone microenvironment sustains survival of myeloma cells. Therefore, a challenge for treating multiple myeloma is discovering drugs targeting not only myeloma cells but also osteoclasts and osteoblasts. Because resveratrol (trans-3,4',5 trihydroxystilbene) is reported to display antitumor activities on a variety of human cancer cells, we investigated the effects of this natural compound on myeloma and bone cells. We found that resveratrol reduces dose-dependently the growth of myeloma cell lines (RPMI 8226 and OPM-2) by a mechanism involving cell apoptosis. In cultures of human primary monocytes, resveratrol inhibits dose dependently receptor activator of nuclear factor-kappaB (NF-kappaB) ligand induced formation of tartrate-resistant acid phosphatase (TRACP)-positive multinucleated cells, TRACP activity in the medium, up-regulation of cathepsin K gene expression, and bone resorption. These inhibitions are associated with a down-regulation of RANK expression at both mRNA and cell surface protein levels and a decrease of NFATc1 stimulation and NF-kappaB nuclear translocation, whereas the gene expression of c-fms, CD14, and CD11a is up-regulated. Finally, resveratrol promotes dose-dependently the expression of osteoblast markers like osteocalcin and osteopontin in human bone marrow mesenchymal stem cells (hMSC TERT) and stimulates their response to 1,25(OH)2 vitamin D3 [1,25(OH)2D3]. Moreover, resveratrol up-regulates dose-dependently the expression of 1,25(OH)2D3 nuclear receptor. Taken together, these results suggest that resveratrol or its derivatives deserve attention as potential drugs for treating multiple myeloma. PMID- 16267020 TI - Mammalian target of rapamycin, a molecular target in squamous cell carcinomas of the head and neck. AB - Emerging knowledge on how the dysregulated function of signaling networks contributes to the malignant growth of squamous cell carcinoma of the head and neck (HNSCC) can now be exploited to identify novel mechanism-based anticancer treatments. In this regard, we have observed that persistent activation of the serine/threonine kinase Akt is a frequent event in HNSCC, and that blockade of its upstream kinase, 3'-phosphoinositide-dependent kinase 1, potently inhibits tumor cell growth. Akt promotes cell proliferation by its ability to coordinate mitogenic signaling with energy- and nutrient-sensing pathways that control protein synthesis through the atypical serine/threonine kinase, mammalian target of rapamycin (mTOR). This kinase, in turn, phosphorylates key eukaryotic translation regulators, including p70-S6 kinase and the eukaryotic translation initiation factor, 4E binding protein 1. Indeed, we show here that aberrant accumulation of the phosphorylated active form of S6, the most downstream target of the Akt-mTOR-p70-S6 kinase pathway, is a frequent event in clinical specimens from patients with HNSCC and their derived cell lines. Of interest, this enhanced level of the phosphorylated active form of S6 was rapidly reduced in HNSCC cell lines and HNSCC xenograft models at clinically relevant doses of rapamycin, which specifically inhibits mTOR. Furthermore, we observed that rapamycin displays a potent antitumor effect in vivo, as it inhibits DNA synthesis and induces the apoptotic death of HNSCC cells, ultimately resulting in tumor regression. These findings identify the Akt-mTOR pathway as a potential therapeutic target for HNSCC, and may provide the rationale for the early clinical evaluation of rapamycin and its analogues in patients with HNSCC. PMID- 16267021 TI - Development of a conditional in vivo model to evaluate the efficacy of small molecule inhibitors for the treatment of Raf-transformed hematopoietic cells. AB - Conditionally active forms of the Raf proteins (Raf-1, B-Raf, and A-Raf) were created by ligating NH2-terminal truncated activated forms (Delta) to the estrogen receptor (ER) hormone-binding domain resulting in estradiol-regulated constructs (DeltaRaf:ER). These different Raf:ER oncoproteins were introduced into the murine FDC-P1 hematopoietic cell line, and cells that grew in response to the three DeltaRaf:ER oncoproteins were isolated. The ability of FDC-P1, DeltaRaf-1:ER, DeltaA-Raf:ER, and DeltaB-Raf:ER cells to form tumors in severe combined immunodeficient mice was compared. Mice injected with DeltaRaf:ER cells were implanted with beta-estradiol pellets to induce the DeltaRaf:ER oncoprotein. Cytokine-dependent parental cell lines did not form tumors. Implantation of beta estradiol pellets into mice injected with DeltaRaf:ER cells significantly accelerated tumor onset and tumor size. The recovered DeltaRaf:ER cells displayed induction of extracellular signal-regulated kinase (ERK) in response to beta estradiol stimulation, indicating that they had retained conditional activation of ERK even when passed through a severe combined immunodeficient mouse. The DeltaRaf:ER cells were very sensitive to induction of apoptosis by the mitogen activated protein/ERK kinase (MEK) 1 inhibitor CI1040 whereas parental cells were much less affected, demonstrating that the MEK1 may be useful in eliminating Ras/Raf/MEK-transformed cells. Furthermore, the effects of in vivo administration of the MEK1 inhibitor were evaluated and this inhibitor was observed to suppress the tumorigenicity of the injected cells. This DeltaRaf:ER system can serve as a preclinical model to evaluate the effects of signal transduction inhibitors which target the Raf and MEK proteins. PMID- 16267022 TI - Mitochondrially targeted p53 has tumor suppressor activities in vivo. AB - Complex proapoptotic functions are essential for the tumor suppressor activity of p53. We recently described a novel transcription-independent mechanism that involves a rapid proapoptotic action of p53 at the mitochondria and executes the shortest known circuitry of p53 death signaling. Here, we examine if this p53 dependent mitochondrial program could be exploited for tumor suppression in vivo. To test this, we engage Emu-Myc transgenic mice, a well-established model of p53 dependent lymphomagenesis. We show that exclusive delivery of p53 to the outer mitochondrial membrane confers a significant growth disadvantage on Emu-Myc transformed B-cells of p53-deficient or alternate reading frame-deficient genotypes, resulting in efficient induction of apoptosis and impinged proliferation. Conversely, normal cells from thymus, spleen, and bone marrow showed poor infectivity with these viruses. This proof-of-principle experiment shows that exclusive reliance on the direct mitochondrial program exerts a significant tumor suppressor activity in vivo. Our in vivo data on the direct mitochondrial apoptotic p53 program lays the groundwork to further investigate its efficacy and safety and to address its possible therapeutic value in the future. PMID- 16267023 TI - Myxoma virus is a novel oncolytic virus with significant antitumor activity against experimental human gliomas. AB - Myxoma virus, a poxvirus previously considered rabbit specific, can replicate productively in a variety of human tumor cells in culture. The purpose of this study was to determine if there was efficacy or toxicities of this oncolytic virus against experimental models of human malignant gliomas in vitro, in vivo, and ex vivo in malignant glioma specimens. In vitro, the majority of glioma cell lines tested (7 of 8, 87.5%) were fully permissive for myxoma virus replication and killed by infection. In vivo, intracerebral (i.c.) myxoma virus inoculation was well tolerated and produced only minimal focal inflammatory changes at the site of viral inoculation. U87 and U251 orthotopic xenograft models were used to assess myxoma virus efficacy in vivo. A single intratumoral injection of myxoma virus dramatically prolonged median survival compared with treatment with UV inactivated myxoma virus. Median survival was not reached in myxoma virus-treated groups versus 47.3 days (U87; P = 0.0002) and 50.7 days (U251; P = 0.0027) in UV inactivated myxoma virus-treated groups. Most myxoma virus-treated animals (12 of 13, 92%) were alive and apparently "cured" when the experiment was finished (>130 days). Interestingly, we found a selective and long-lived myxoma virus infection in gliomas in vivo. This is the first demonstration of the oncolytic activity of myxoma virus in vivo. The nonpathogenic nature of myxoma virus outside of the rabbit host, its capacity to be genetically modified, its ability to produce a long-lived infection in human tumor cells, and the lack of preexisting antibodies in the human population suggest that myxoma virus may be an attractive oncolytic agent against human malignant glioma. PMID- 16267025 TI - Doxorubicin induces apoptosis in germ line stem cells in the immature rat testis and amifostine cannot protect against this cytotoxicity. AB - The underlying primary damage to the seminiferous epithelium caused by chemotherapeutic regimens at childhood is largely unknown. The present investigation was designed to identify acute cytotoxic events in the testis caused by a single dose of doxorubicin. Male rats at 6, 16, and 24 days of age were injected with doxorubicin (3 mg/kg, i.p.) or vehicle (saline) alone and 24 and 48 hours later, the germ cell types and apoptotic cells in the seminiferous epithelium were examined. As indicated by microscopy and terminal deoxyribonucleotidyl transferase-mediated dUTP nick end labeling staining, an 8 fold increase in the number of apoptotic germ cells in the testes of 6-day-old rats was observed 48 hours after doxorubicin treatment. Spermatogonia migrating to the basement membrane were the primary cell type undergoing this induced apoptosis. A single dose of amifostine (200 mg/kg) administered i.p. 15 minutes before injection of doxorubicin provided no protection against this enhanced apoptosis. Under the same conditions, testicular levels of p53 and activated caspase 8 were elevated, whereas the level of murine double minute-2 was lowered. In contrast, doxorubicin treatment did not result in any significant change in the physiologic, stage-specific germ cell apoptosis occurring in the testes of 16 and 24-day-old rats. These observations suggest that the initiation phase of spermatogenesis is highly sensitive to doxorubicin-induced apoptosis. Gonocytes and early spermatogonia are the cell types that are vulnerable to this p53 trigged apoptosis, which results in a decrease in the size of the pool of germ line stem cells. Amifostine fails to protect the germ cells against this cytotoxic insult. PMID- 16267024 TI - The enhanced tumor selectivity of an oncolytic vaccinia lacking the host range and antiapoptosis genes SPI-1 and SPI-2. AB - The ability of cancer cells to evade apoptosis may permit survival of a recombinant vaccinia lacking antiapoptotic genes in cancer cells compared with normal cells. We have explored the deletion of two vaccinia virus host range/antiapoptosis genes, SPI-1 and SPI-2, for their effects on the viral replication and their ability to induce cell death in infected normal and transformed cells in vitro. Indeed, in three paired normal and transformed cell types, the SPI-1 and SPI-2 gene-deleted virus (vSP) preferentially replicates in transformed cells or p53-null cells when compared with their normal counterparts. This selectivity may be derived from the fact that vSP-infected normal cells died faster than infected cancer cells. A fraction of infected cells died with evidence of necrosis as shown by both flow cytometry and detection of high mobility group B1 protein released from necrotic cells into the culture supernatant. When administered to animals, vSP retains full ability to replicate in tumor tissues, whereas replication in normal tissues is greatly diminished. In a model of viral pathogenesis, mice treated with vSP survived substantially longer when compared with mice treated with the wild-type virus. The mutant virus vSP displayed significant antitumoral effects in an MC38 s.c. tumor model in both nude (P < 0.001) and immunocompetent mice (P < 0.05). We conclude that this recombinant vaccinia vSP shows promise for oncolytic virus therapy. Given its enhanced tumor selectivity, improved safety profile, and substantial oncolytic effects following systemic delivery in murine models, it should also serve as a useful vector for tumor-directed gene therapy. PMID- 16267026 TI - Formation of 17-allylamino-demethoxygeldanamycin (17-AAG) hydroquinone by NAD(P)H:quinone oxidoreductase 1: role of 17-AAG hydroquinone in heat shock protein 90 inhibition. AB - We have examined the role of NAD(P)H:quinone oxidoreductase 1 (NQO1) in the bioreductive metabolism of 17-allylamino-demethoxygeldanamycin (17-AAG). High performance liquid chromatography (HPLC) analysis of the metabolism of 17-AAG by recombinant human NQO1 revealed the formation of a more polar metabolite 17 AAGH2. The formation of 17-AAGH2 was NQO1 dependent, and its formation could be inhibited by the addition of 5-methoxy-1,2-dimethyl-3-[(4 nitrophenoxy)methyl]indole-4,7-dione (ES936), a mechanism-based (suicide) inhibitor of NQO1. The reduction of 17-AAG to the corresponding hydroquinone 17 AAGH2 was confirmed by tandem liquid chromatography-mass spectrometry. 17-AAGH2 was relatively stable and only slowly underwent autooxidation back to 17-AAG over a period of hours. To examine the role of NQO1 in 17-AAG metabolism in cells, we used an isogenic pair of human breast cancer cell lines differing only in NQO1 levels. MDA468 cells lack NQO1 due to a genetic polymorphism, and MDA468/NQ16 cells are a stably transfected clone that express high levels of NQO1 protein. HPLC analysis of 17-AAG metabolism using cell sonicates and intact cells showed that 17-AAGH2 was formed by MDA468/NQ16 cells, and formation of 17-AAGH2 could be inhibited by ES936. No 17-AAGH2 was detected in sonicates or intact MDA468 cells. Following a 4-hour treatment with 17-AAG, the MDA468/NQ16 cells were 12-fold more sensitive to growth inhibition compared with MDA468 cells. More importantly, the increased sensitivity of MDA468/NQ16 cells to 17-AAG could be abolished if the cells were pretreated with ES936. Cellular markers of heat shock protein (Hsp) 90 inhibition, Hsp70 induction, and Raf-1 degradation were measured by immunoblot analysis. Marked Hsp70 induction and Raf-1 degradation was observed in MDA468/NQ16 cells but not in MDA468 cells. Similarly, downstream Raf-1 signaling molecules mitogen-activated protein kinase/extracellular signal-regulated kinase (ERK) kinase and ERK also showed decreased levels of phosphorylation in MDA468/NQ16 cells but not in MDA468 cells. The ability of 17-AAG and 17-AAGH2 to inhibit purified yeast and human Hsp90 ATPase activity was examined. Maximal 17 AAG-induced ATPase inhibition was observed in the presence of NQO1 and could be abrogated by ES936, showing that 17-AAGH2 was a more potent Hsp90 inhibitor compared with 17-AAG. Molecular modeling studies also showed that due to increased hydrogen bonding between the hydroquinone and the Hsp90 protein, 17 AAGH2 was bound more tightly to the ATP-binding site in both yeast and human Hsp90 models. In conclusion, these studies have shown that reduction of 17-AAG by NQO1 generates 17-AAGH2, a relatively stable hydroquinone that exhibits superior Hsp90 inhibition. PMID- 16267027 TI - N-benzyladriamycin-14-valerate (AD198) induces apoptosis through protein kinase C delta-induced phosphorylation of phospholipid scramblase 3. AB - Phospholipid scramblase 3 (PLS3) is an enzyme that plays a critical role in mitochondrial morphology, functions, and apoptotic response. During apoptosis, activated protein kinase C-delta (PKC-delta) translocates to mitochondria and phosphorylates PLS3. Here, we utilize an extranuclear-targeted anthracycline N benzyladriamycin-14-valerate (AD198), a PKC-delta activator, to investigate the mechanism of PLS3 phosphorylation by PKC-delta. Overexpression of PLS3 enhanced, whereas down-regulation of PLS3 by small interfering RNA decreased, the sensitivity of AD198-induced apoptosis. Overexpression of PKC-delta, but not the kinase-defective PKC-delta, and AD198 treatment enhanced threonine phosphorylation of PLS3. The phosphorylated threonine was mapped to Thr21 of PLS3. Mutation of Thr21 to alanine did not affect mitochondrial localization of PLS3 but abolished threonine phosphorylation by PKC-delta in vitro and AD198 induced PLS3 phosphorylation in vivo. Expression of PLS3(T21A) in cells could not enhance AD198-induced apoptosis compared with expression of the wild-type PLS3. Using benzyloxycarbonyl-Val-Ala-Asp-(OMe) fluoromethyl ketone and cyclosporine A, we also showed that AD198-induced PLS3 phosphorylation occurs upstream of caspase activation and independent of mitochondrial permeability transition. These studies establish that AD198-activated PKC-delta induces phosphorylation of mitochondrial PLS3 at Thr21 and that PLS3 is a critical downstream effector of PKC-delta in AD198-induced apoptosis. PMID- 16267028 TI - Epigenetic regulation of WTH3 in primary and cultured drug-resistant breast cancer cells. AB - Previous studies showed that the WTH3 gene functioned as a negative regulator during multidrug resistance (MDR) development in vitro. To understand whether this gene is also involved in clinical drug resistance, hypermethylation at its promoter region observed in cultured MDR MCF7/AdrR cells was examined in primary drug-resistant breast cancer epithelial cells isolated from effusions of breast cancer patients. The results showed that this event also occurred in drug resistant breast cancer epithelial cells and a newly induced drug-resistant cell line, MCF7/inR. Interestingly, we found that a CpG (CpG 23) that was close to the TATA-like box was constantly methylated in the WTH3 promoter of drug-resistant breast cancer epithelial and cultured MDR cells. Mutagenic study suggested that this CpG site had a functional effect on promoter activity. We also discovered that MCF7/AdrR cells treated with trichostatin A, a histone deacetylase inhibitor, exhibited higher WTH3, but lower MDR1, expression. A reverse correlation between WTH3 and MDR1 gene expression was also observed in MCF7/AdrR, and its non-MDR parental cell line, MCF7/WT. This result indicated that both DNA methylation and histone deacetylase could act in concert to inhibit WTH3 and consequently stimulate MDR1 expression. This hypothesis was supported by data obtained from introducing the WTH3 transgene into MDR cell lines, which reduced endogenous MDR1 expression. Therefore, our studies suggested that the behavior of WTH3 in primary drug-resistant breast cancer epithelial cells was similar to that in a model system where epigenetic regulation of the WTH3 gene was linked to the MDR phenotype. PMID- 16267029 TI - Renal cancer treatment with low levels of mixed chimerism induced by nonmyeloablative regimen using cyclophosphamide in mice. AB - Recently, much attention has been paid to nonmyeloablative allogeneic stem cell transplantation for the treatment of metastatic renal cancer. Mature donor T cells cause graft-versus-host disease (GVHD) although they are also the main mediators of the beneficial graft-versus-tumor activity associated with this treatment. Hence, the segregation of the graft-versus-tumor activity from GVHD is an important challenge in managing the clinical course of treatment. We previously reported a series of studies regarding the allograft tolerance induced by allogeneic spleen cells (with bone marrow cells) and cyclophosphamide in mice. Here, we show a modified cyclophosphamide-induced tolerance system for the treatment of murine renal cell carcinoma, RENCA, by shifting the equal balance between graft-versus-host and host-versus-graft reactions toward graft-versus host reaction with donor lymphocyte infusion. Our results clearly show the antitumor activity against RENCA with only low levels of mixed chimerism in the periphery and the in vivo and in vitro acquired immunity against RENCA even when mixed chimerism is almost undetectable. Because the withdrawal of mixed chimerism reduces the risk of GVHD, the antitumor activity is thus sequentially segregated from the initial GVHD in our model. We believe that this is the first unique model system of nonmyeloablative allogeneic hemopoietic cell transplantation to ever be reported for the treatment of renal cancer. PMID- 16267030 TI - Protein transduction of dendritic cells for NY-ESO-1-based immunotherapy of myeloma. AB - Myeloma vaccines, based on dendritic cells pulsed with idiotype or tumor lysate, have been met with limited success, probably in part due to insufficient cross priming of myeloma antigens. A powerful method to introduce myeloma-associated antigens into the cytosol of dendritic cells is protein transduction, a process by which proteins fused with a protein transduction domain (PTD) freely traverse membrane barriers. NY-ESO-1, an immunogenic antigen by itself highly expressed in 60% of high-risk myeloma patients, was purified to near homogeneity both alone and as a recombinant fusion protein with a PTD, derived from HIV-Tat. Efficient entry of PTD-NY-ESO-1 into dendritic cells, confirmed by microscopy, Western blotting, and intracellular flow cytometry, was achieved without affecting dendritic cell phenotype. Experiments with amiloride, which inhibits endocytosis, and N-acetyl-l-leucinyl-l-norleucinal, a proteasome inhibitor, confirmed that PTD NY-ESO-1 entered dendritic cells by protein transduction and was degraded by the proteasome. Tetramer analysis indicated superior generation of HLA-A2.1, CD8+ T lymphocytes specific for NY-ESO-1(157-165) with PTD-NY-ESO-1 compared with NY-ESO 1 control protein (44% versus 2%, respectively). NY-ESO-1-specific T lymphocytes generated with PTD-NY-ESO-1 secreted IFN-gamma indicative of a Tc1-type cytokine response. Thus, PTD-NY-ESO-1 accesses the cytoplasm by protein transduction, is processed by the proteasome, and NY-ESO-1 peptides presented by HLA class I elicit NY-ESO-1-specific T lymphocytes. PMID- 16267031 TI - Gene expression profiling identifies BAX-delta as a novel tumor antigen in acute lymphoblastic leukemia. AB - The identification of new tumor-associated antigens (TAA) is critical for the development of effective immunotherapeutic strategies, particularly in diseases like B-cell acute lymphoblastic leukemia (B-ALL), where few target epitopes are known. To accelerate the identification of novel TAA in B-ALL, we used a combination of expression profiling and reverse immunology. We compared gene expression profiles of primary B-ALL cells with their normal counterparts, B-cell precursors. Genes differentially expressed by B-ALL cells included many previously identified as TAA in other malignancies. Within this set of overexpressed genes, we focused on those that may be functionally important to the cancer cell. The apoptosis-related molecule, BAX, was highly correlated with the ALL class distinction. Therefore, we evaluated BAX and its isoforms as potential TAA. Peptides from the isoform BAX-delta bound with high affinity to HLA-A*0201 and HLA-DR1. CD8+ CTLs specific for BAX-delta epitopes or their heteroclitic peptides could be expanded from normal donors. BAX-delta-specific T cells lysed peptide-pulsed targets and BAX-delta-expressing leukemia cells in a MHC-restricted fashion. Moreover, primary B-ALL cells were recognized by BAX delta-specific CTL, indicating that this antigen is naturally processed and presented by tumor cells. This study suggests that (a) BAX-delta may serve as a widely expressed TAA in B-ALL and (b) gene expression profiling can be a generalizable tool to identify immunologic targets for cancer immunotherapy. PMID- 16267032 TI - Tumor cell loaded type-1 polarized dendritic cells induce Th1-mediated tumor immunity. AB - Dendritic cells are professional antigen-presenting cells capable of inducing and regulating innate and antigen-specific immune responses. Therapeutic cancer vaccines using ex vivo engineered or in vivo targeted dendritic cells are being evaluated in clinical trials. T-helper type-1 (Th1)-skewed immune responses are characterized by the preferential induction of antigen-specific IFN-gamma secreting CD4+ T cells and correlate with effector mechanisms important for tumor and viral immunity. Methods to "polarize" human monocyte-derived dendritic cells for the preferential induction of Th1-skewed immune responses have been developed, and polarized dendritic cells (DC1s) are being evaluated in preclinical and clinical studies. Here, we show that stimulation of bone marrow derived murine dendritic cell populations with poly(I:C) and CpGs results in phenotypic maturation of dendritic cells and synergistic induction of durable, high-level IL-12p70 secretion characteristic of human type-1 polarized dendritic cells. Functionally, these dendritic cells induce antigen-specific Th1-type CD4+ T-cell activation in vitro and in vivo. Dendritic cell maturation and polarization are not inhibited by the presence of live B16 melanoma tumor cells, and tumor-loaded DC1s induce delayed-type hypersensitivity responses in vivo. DC1s loaded with B16 melanoma cells and injected into tumor-bearing mice induce Th1-skewed tumor-specific CD4+ T cells and a significant reduction in tumor growth. Tumor infiltrates in DC1-immunized animals are characterized by the presence of CD4+ T cells and activated macrophages. These results show a murine model of DC1 function and suggest an important role for CD4+ T cells and macrophages in DC1-induced antitumor immune responses. They have implications for the future development of DC1-based immunotherapies and strategies for clinical immune monitoring of their effectiveness. PMID- 16267033 TI - A novel strategy for the discovery of MHC class II-restricted tumor antigens: identification of a melanotransferrin helper T-cell epitope. AB - CD4+ helper T cells play a critical role in orchestrating host immune responses, including antitumor immunity. The limited availability of MHC class II-associated tumor antigens is still viewed as a major obstacle in the use of CD4+ T cells in cancer vaccines. Here, we describe a novel approach for the identification of MHC class II tumor-associated antigens (TAAs). By combining two-dimensional liquid chromatography and nanoelectrospray ionization tandem mass spectrometry, we developed a highly sensitive method for the detection of human leukocyte antigen (HLA)-DR-associated peptides of dendritic cells upon exposure to necrotic tumor cells. This approach led to the identification of a novel MHC class II-restricted TAA epitope derived from melanotransferrin. The epitope stimulated T cells derived from melanoma patients and healthy individuals and displayed promiscuity in HLA-DR restriction. Moreover, the same peptide was also presented by MHC class II-positive melanoma cells. This strategy may contribute to increase the number of tumor epitopes presented by MHC class II molecules and may support the development of more efficacious vaccines against cancer. PMID- 16267034 TI - High frequencies of functional tumor-reactive T cells in bone marrow and blood of pancreatic cancer patients. AB - Pancreatic cancer is characterized by aggressive growth and treatment resistance. New approaches include immunotherapeutic strategies but the type and extent of spontaneous immune responses against tumor antigens remains unclear. A dominance of TH2 cytokines in patients' sera reported previously suggests systemic tumor induced immunosuppression, potentially inhibiting the induction of tumor-reactive T cells. We characterized the localization, frequencies, and functional potential of spontaneously induced memory T cells specific for individual tumor antigens or the tumor-associated antigen mucin-1 in the peripheral blood and bone marrow of 41 pancreatic cancer patients. We found high numbers of tumor-reactive T cells in all bone marrow samples and in 50% of the blood samples. These cells secreted the TH1 cytokine IFN-gamma rather than TH2 cytokines upon stimulation with tumor antigens. Although consistently induced during pancreatic cancer, T cells specific for pancreatic antigens were not detected during chronic pancreatitis, suggesting that their evaluation may be of diagnostic use in both diseases. Freshly isolated T cells from cancer patients recognized autologous tumor cells and rejected them in vitro and in a xenotransplant model in vivo, suggesting their therapeutic potential. Thus, tumor antigen-specific T cell responses occur regularly during pancreatic cancer disease and lead to enrichment of tumor cell reactive memory T cells in the bone marrow. The bone marrow can therefore be considered an important organ for antitumor immune responses in pancreatic cancer. PMID- 16267035 TI - Genetic ablation of protein tyrosine phosphatase 1B accelerates lymphomagenesis of p53-null mice through the regulation of B-cell development. AB - Protein tyrosine phosphatase 1B (PTP1B) is involved in multiple signaling pathways by down-regulating several tyrosine kinases. For example, gene-targeting studies in mice have established PTP1B as a critical physiologic regulator of metabolism by attenuating insulin signaling. PTP1B is an important target for the treatment of diabetes, because the PTP1B null mice are resistant to diet-induced diabetes and obesity. On the other hand, despite the potential for enhanced oncogenic signaling in the absence of PTP1B, PTP1B null mice do not develop spontaneous tumors. Because the majority of human cancers harbor mutations in p53, we generated p53/PTP1B double null mice to elucidate the role of PTP1B in tumorigenesis. We show that genetic ablation of PTP1B in p53 null mice decreases survival rate and increases susceptibility towards the development of B lymphomas. This suggested a role for PTP1B in lymphopoiesis, and we report that PTP1B null mice have an accumulation of B cells in bone marrow and lymph nodes, which contributed to the increased incidence of B lymphomas. The mean time of tumor development and tumor spectrum are unchanged in p53-/-PTP1B+/- mice. We conclude that PTP1B is an important determinant of the latency and type of tumors in a p53-deficient background through its role in the regulation of B-cell development. PMID- 16267036 TI - Application of embryonic lethal or other obvious phenotypes to characterize the clinical significance of genetic variants found in trans with known deleterious mutations. AB - This work describes an approach to characterize the clinical significance of genetic variants detected during the genetic testing of BRCA1 in patients from hereditary breast/ovarian cancer families. Results from transgenic mice and extensive clinical testing support the hypothesis that biallelic BRCA1 mutations result in embryonic lethality. Therefore, it is reasonable to conclude that variants of uncertain clinical significance found to reside in trans with known deleterious mutations impart reduced risk for cancer. This approach was applied to a large data set of 55,630 patients who underwent clinical BRCA1 screening by whole gene direct DNA sequencing. Fourteen common single nucleotide polymorphisms (SNPs) were used to assign 10 previously defined common, recurrent, or canonical haplotypes in 99% of these cases. From a total of 1,477 genetic variants detected in these patients, excluding haplotype-tagging SNPs, 877 (59%) could be unambiguously assigned to one or more haplotypes. In 41 instances, variants previously classified as being of uncertain clinical significance, mostly missense variants, were excluded as fully penetrant mutations due to their coincidence in trans with known deleterious mutations. From a total of 1,150 patients that harbored these 41 variants, 956 carried one as the sole variant of uncertain clinical significance reported. This approach could have widespread application to other disease genes where compound heterozygous mutations are incompatible with life or result in obvious phenotypes. This largely computational technique is advantageous because it relies upon existing clinical data and is likely to prove informative for prevalent genetic variants in large data sets. PMID- 16267037 TI - Quantification of cellular proliferation in tumor and normal tissues of patients with breast cancer by [18F]fluorothymidine-positron emission tomography imaging: evaluation of analytical methods. AB - There is an unmet need to develop imaging methods for the early and objective assessment of breast tumors to therapy. 3'-Deoxy-3'-[18F]fluorothymidine ([18F]FLT)-positron emission tomography represents a new approach to imaging thymidine kinase activity, and hence, cellular proliferation. We compared graphical, spectral, and semiquantitative analytic methodologies for quantifying [18F]FLT kinetics in tumor and normal tissue of patients with locally advanced and metastatic breast cancer. The resultant kinetic parameters were correlated with the Ki-67 labeling index from tumor biopsies. [18F]FLT accumulation was detected in primary tumor, nodal disease, and lung metastasis. In large tumors, there was substantial heterogeneity in regional radiotracer uptake, reflecting heterogeneity in cellular proliferation; radiotracer uptake in primary tumors also differed from that of metastases. [18F]FLT was metabolized in patients to a single metabolite [18F]FLT-glucuronide. Unmetabolized [18F]FLT accounted for 71.54 +/- 1.50% of plasma radioactivity by 90 minutes. The rate constant for the metabolite-corrected net irreversible uptake of [18F]FLT (Ki) ranged from 0.6 to 10.4 x 10(-4) and from 0 to 0.6 x 10(-4) mL plasma cleared/s/mL tissue in tumor (29 regions, 15 patients) and normal tissues, respectively. Tumor Ki and fractional retention of radiotracer determined by spectral analysis correlated with Ki-67 labeling index (r = 0.92, P < 0.0001 and r = 0.92, P < 0.0001, respectively). These correlations were superior to those determined by semiquantitative methods. We conclude that [18F]FLT-positron emission tomography is a promising clinical tool for imaging cellular proliferation in breast cancer, and is most predictive when analyzed by graphical and spectral methods. PMID- 16267039 TI - SV40 detection in human tumor specimens. PMID- 16267038 TI - HER2/neu-induced mammary tumorigenesis and angiogenesis are reduced in cyclooxygenase-2 knockout mice. AB - The inducible prostaglandin synthase cyclooxygenase-2 (Cox-2) is overexpressed in approximately 40% of human breast cancers and at higher frequencies in preinvasive ductal carcinoma in situ (DCIS). Cox-2 expression is particularly associated with overexpression of human epidermal growth factor receptor 2 (HER2/neu). To definitively interrogate the role of Cox-2 in mammary neoplasia, we have used a genetic approach, crossing Cox-2-deficient mice with a HER2/neu transgenic strain, MMTV/NDL. At 20 weeks of age, mammary glands from virgin MMTV/NDL females contained multiple focal tumors, or mammary intraepithelial neoplasias, which histologically resembled human DCIS. Mammary tumor multiplicity and prostaglandin E2 (PGE2) levels were significantly decreased in Cox-2 heterozygous and knockout animals relative to Cox-2 wild-type controls. Notably, the proportion of larger tumors was decreased in Cox-2-deficient mice. HER2/neu induced mammary hyperplasia was also substantially reduced in Cox-2 null mice. Additionally, mammary glands from Cox-2 knockout mice exhibited a striking reduction in vascularization, and expression of proangiogenic genes was correspondingly reduced. Decreased vascularization was observed both in dysplastic and normal-appearing regions of Cox-2-null mammary glands. Our data provide the first genetic evidence that Cox-2 contributes to HER2/neu-induced mammary tumorigenesis. This finding may help to explain the reduced risk of breast cancer associated with regular use of nonsteroidal anti-inflammatory drugs. PMID- 16267040 TI - Structure of the pyrroloquinoline quinone radical in quinoprotein ethanol dehydrogenase. AB - Quinoprotein alcohol dehydrogenases use the pyrroloquinoline quinone (PQQ) cofactor to catalyze the oxidation of alcohols. The catalytic cycle is thought to involve a hydride transfer from the alcohol to the oxidized PQQ, resulting in the generation of aldehyde and reduced PQQ. Reoxidation of the cofactor by cytochrome proceeds in two sequential steps via the PQQ radical. We have used a combination of electron nuclear double resonance and density functional theory to show that the PQQ radical is not protonated at either O-4 or O-5, a result that is at variance with the general presumption of a singly protonated radical. The quantum mechanical calculations also show that reduced PQQ is unlikely to be protonated at O-5; rather, it is either singly protonated at O-4 or not protonated at either O-4 or O-5, a result that also challenges the common assumption of a reduced PQQ protonated at both O-4 and O-5. The reaction cycle of PQQ-dependent alcohol dehydrogenases is revised in light of these findings. PMID- 16267041 TI - The role of the epsilon subunit in the Escherichia coli ATP synthase. The C terminal domain is required for efficient energy coupling. AB - The role of the C-domain of the epsilon subunit of ATP synthase was investigated by fusing either the 20-kDa flavodoxin (Fd) or the 5-kDa chitin binding domain (CBD) to the N termini of both full-length epsilon and a truncation mutant epsilon(88-stop). All mutant epsilon proteins were stable in cells and supported F1F0 assembly. Cells expressing the Fd-epsilon or Fd-epsilon(88-stop) mutants were unable to grow on acetate minimal medium, indicating their inability to carry out oxidative phosphorylation because of steric blockage of rotation. The other forms of epsilon supported growth on acetate. Membrane vesicles containing Fd-epsilon showed 23% of the wild type ATPase activity but no proton pumping, suggesting that the ATP synthase is intrinsically partially uncoupled. Vesicles containing CBD-epsilon were indistinguishable from the wild type in ATPase activity and proton pumping, indicating that the N-terminal fusions alone do not promote uncoupling. Fd-epsilon(88-stop) caused higher rates of uncoupled ATP hydrolysis than Fd-epsilon, and epsilon(88-stop) showed an increased rate of membrane-bound ATP hydrolysis but decreased proton pumping relative to the wild type. Both results demonstrate the role of the C-domain in coupling. Analysis of the wild type and epsilon(88-stop) mutant membrane ATPase activities at concentrations of ATP from 50 mum to 8 mm showed no significant dependence of the ratio of bound/released ATPase activity on ATP concentration. These results support the hypothesis that the main function of the C-domain in the Escherichia coli epsilon subunit is to reduce uncoupled ATPase activity, rather than to regulate coupled activity. PMID- 16267042 TI - Site-specific monoubiquitination of IkappaB kinase IKKbeta regulates its phosphorylation and persistent activation. AB - Transcription factor NF-kappaB governs the expression of multiple genes involved in cell growth, immunity, and inflammation. Nuclear translocation of NF-kappaB is regulated from the cytoplasm by IkappaB kinase-beta (IKKbeta), which earmarks inhibitors of NF-kappaB for polyubiquination and proteasome-mediated degradation. Activation of IKKbeta is contingent upon signal-induced phosphorylation of its T loop at Ser-177/Ser-181. T loop phosphorylation also renders IKKbeta a substrate for monoubiquitination in cells exposed to chronic activating cues, such as the Tax oncoprotein or sustained signaling through proinflammatory cytokine receptors. Here we provide evidence that the T loop-proximal residue Lys-163 in IKKbeta serves as a major site for signal-induced monoubiquitination with significant regulatory potential. Conservative replacement of Lys-163 with Arg yielded a monoubiquitination-defective mutant of IKKbeta that retains kinase activity in Tax-expressing cells but is impaired for activation mediated by chronic signaling from the type 1 receptor for tumor necrosis factor-alpha. Phosphopeptide mapping experiments revealed that the Lys-163 --> Arg mutation also interferes with proper in vivo but not in vitro phosphorylation of cytokine responsive serine residues located in the distal C-terminal region of IKKbeta. Taken together, these data indicate that chronic phosphorylation of IKKbeta at Ser-177/Ser-181 leads to monoubiquitin attachment at nearby Lys-163, which in turn modulates the phosphorylation status of IKKbeta at select C-terminal serines. This mechanism for post-translational cross-talk may play an important role in the control of IKKbeta signaling during chronic inflammation. PMID- 16267043 TI - Deregulation of proteasome function induces Abl-mediated cell death by uncoupling p130CAS and c-CrkII. AB - Cell migration and survival are coordinately regulated through activation of c Abl (Abl) family tyrosine kinases. Activated Abl phosphorylates tyrosine 221 of c CrkII (Crk; Crk-Y221-P), which prevents Crk from binding to the docking protein p130(CAS) (CAS). Disruption of CAS-Crk binding blocks downstream effectors of the actin cytoskeleton and focal adhesion assembly, inhibits cell migration, and disrupts survival signals leading to apoptosis. Here we show that inhibition of the 26 S proteasome and ubiquitination facilitates Abl-mediated Crk-Y221-P, leading to disassembly of CAS-Crk complexes in cells. Surprisingly, inhibition of these molecular interactions does not perturb cell migration but rather specifically induces apoptosis. Furthermore, we demonstrate that attachment to an extracellular matrix plays a key role in regulating the apoptotic machinery through caspase-mediated cleavage of Abl and Crk-Y221-P. Our findings indicate that regulated protein degradation by the proteasome specifically controls cell death through regulation of Abl-mediated Crk Tyr221 phosphorylation and assembly of the CAS-Crk signaling scaffold. PMID- 16267044 TI - The plant plasma membrane Ca2+ pump ACA8 contains overlapping as well as physically separated autoinhibitory and calmodulin-binding domains. AB - In plant Ca(2+) pumps belonging to the P(2B) subfamily of P-type ATPases, the N terminal cytoplasmic domain is responsible for pump autoinhibition. Binding of calmodulin (CaM) to this region results in pump activation but the structural basis for CaM activation is still not clear. All residues in a putative CaM binding domain (Arg(43) to Lys(68)) were mutagenized and the resulting recombinant proteins were studied with respect to CaM binding and the activation state. The results demonstrate that (i) the binding site for CaM is overlapping with the autoinhibitory region and (ii) the autoinhibitory region comprises significantly fewer residues than the CaM-binding region. In a helical wheel projection of the CaM-binding domain, residues involved in autoinhibition cluster on one side of the helix, which is proposed to interact with an intramolecular receptor site in the pump. Residues influencing CaM negatively are situated on the other face of the helix, likely to face the cytosol, whereas residues controlling CaM binding positively are scattered throughout. We propose that early CaM recognition is mediated by the cytosolic face and that CaM subsequently competes with the intramolecular autoinhibitor in binding to the other face of the helix. PMID- 16267045 TI - Src activation is not necessary for transforming growth factor (TGF)-beta mediated epithelial to mesenchymal transitions (EMT) in mammary epithelial cells. PP1 directly inhibits TGF-beta receptors I and II. AB - Epithelial to mesenchymal transitions (EMTs) are key events during embryonic development and cancer progression. It has been proposed that Src plays a major role in some EMT models, as shown by the overexpression of viral Src (v-Src) in epithelial cells. It is clear that Src family kinases can regulate the integrity of both adherens junctions and focal adhesions; however, their significance in EMT, especially in the physiological context, remains to be elucidated. Here we showed that Src is activated in transforming growth factor-beta1 (TGF-beta1) mediated EMT in mammary epithelial cells and that the Src family kinase inhibitor, PP1, prevents EMT. However, neither a more specific Src family kinase inhibitor, SU6656, nor a dominant-negative Src inhibited TGF-beta1-mediated EMT, leading us to speculate that Src activation is not an essential component of TGF beta1-mediated EMT. Unexpectedly, PP1 prevented Smad2/3 activation by TGF-beta1, whereas SU6656 did not. Most interestingly, an in vitro kinase assay showed that PP1 strongly inhibited the TGF-beta receptor type I, and to a lesser extent, the TGF-beta receptor type II. Taken together, our data indicated that PP1 interferes with TGF-beta1-mediated EMT not by inhibiting Src family kinases but by inhibiting the Smad pathway via a direct inhibition of TGF-beta receptor kinase activity. PMID- 16267046 TI - A structural perspective on enzymes activated by monovalent cations. AB - Enzymes activated by monovalent cations are abundantly represented in plants and the animal world. They have evolved to exploit Na+ and K+, readily available in biological environments, as major driving forces for substrate binding and catalysis. Recent progress in the structural biology of such enzymes has answered long standing questions about the molecular mechanism of activation and the origin of monovalent cation selectivity. That enables a simple classification of these functionally diverse enzymes and reveals unanticipated connections with ion transporters. PMID- 16267047 TI - Two isoforms of a divalent metal transporter (DMT1) in Schistosoma mansoni suggest a surface-associated pathway for iron absorption in schistosomes. AB - We describe two homologues of the mammalian divalent metal transporter (DMT1) for Schistosoma mansoni, a pathogenic intravascular parasite of humans. Schistosomes have a high nutritional and metabolic demand for iron. Nucleotide sequences of the parasite homologues, designated SmDMT1A and -B, are identical in all but the 5'-regions. The predicted amino acid sequences share at least 60% identity with DMT1 (=Nramp2) of humans, mice, and rats, and at least 55% identity with Nramp1 from mice, humans and Caenorhabditis elegans. SmDMT1A is expressed in differentiating eggs, miracidia, cercariae, schistosomula, and adults, whereas SmDMT1B is expressed in all but the miracidium and occurs at lower levels than SmDMT1A in differentiating eggs and cercariae. An iron-responsive element, present at the 3'-untranslated region of many DMT1 molecules, is not present in schistosome mRNAs studied here. A Western blot analysis of adult worm preparations using a homologous rabbit serum raised against a schistosome DMT1 peptide and a heterologous serum raised against mammalian DMT1, revealed a band approximating 115 kDa. By immunofluorescence microscopy, the schistosome DMT1s localize primarily to the tegument. Iron uptake assays demonstrated that SmDMT1s were able to rescue yeast growth in ferrous iron-transport deficient yeast (fet3fet4). The results suggest that schistosomes express molecules for ferrous iron transport in their tegument, suggesting trans-tegumental transport as one means of iron acquisition for these parasites. PMID- 16267048 TI - Genetic ablation of polysialic acid causes severe neurodevelopmental defects rescued by deletion of the neural cell adhesion molecule. AB - Poly-alpha2,8-sialic acid (polySia) is a unique modification of the neural cell adhesion molecule, NCAM, tightly associated with neural development and plasticity. However, the vital role attributed to this carbohydrate polymer has been challenged by the mild phenotype of mice lacking polySia due to NCAM deficiency. To dissect polySia and NCAM functions, we generated polySia-negative but NCAM-positive mice by simultaneous deletion of the two polysialyltransferase genes, St8sia-II and St8sia-IV. Beyond features shared with NCAM-null animals, a severe phenotype with specific brain wiring defects, progressive hydrocephalus, postnatal growth retardation, and precocious death was observed. These drastic defects were selectively rescued by additional deletion of NCAM, demonstrating that they originate from a gain of NCAM functions because of polySia deficiency. The data presented in this study reveal that the essential role of polySia resides in the control and coordination of NCAM interactions during mouse brain development. Moreover, this first demonstration in vivo that a highly specific glycan structure is more important than the glycoconjugate as a whole provides a novel view on the relevance of protein glycosylation for the complex process of building the vertebrate brain. PMID- 16267049 TI - Estrogen-related receptor alpha is a repressor of phosphoenolpyruvate carboxykinase gene transcription. AB - The orphan nuclear receptor estrogen-related receptor (ERR) alpha is a downstream effector of the transcriptional coactivator PGC-1alpha in the regulation of genes important for mitochondrial oxidative capacity. PGC-1alpha is also a potent activator of the transcriptional program required for hepatic gluconeogenesis, and in particular of the key gluconeogenic enzyme phosphoenolpyruvate carboxykinase (PEPCK). We report here that the regulatory sequences of the PEPCK gene harbor a functional ERRalpha binding site. However, in contrast to the co stimulating effects of ERRalpha and PGC-1alpha on mitochondrial gene expression, ERRalpha acts as a transcriptional repressor of the PEPCK gene. Suppression of ERRalpha expression by small interfering RNA leads to reduced binding of ERRalpha to the endogenous PEPCK gene, and an increase in promoter occupancy by PGC 1alpha, suggesting that part of the ERRalpha function at this gene is to antagonize the action of PGC-1alpha. In agreement with the in vitro studies, animals that lack ERRalpha show increased expression of gluconeogenic genes, including PEPCK and glycerol kinase, but decreased expression of mitochondrial genes, such as ATP synthase subunit beta and cytochrome c-1. Our findings suggest that ERRalpha has opposing effects on genes important for mitochondrial oxidative capacity and gluconeogenesis. The different functions of ERRalpha in the regulation of these pathways suggest that enhancing ERRalpha activity could have beneficial effects on glucose metabolism in diabetic subjects by two distinct mechanisms: increasing mitochondrial oxidative capacity in peripheral tissues and liver, and suppressing hepatic glucose production. PMID- 16267050 TI - Expression patterns and post-translational modifications associated with mammalian histone H3 variants. AB - Covalent histone modifications and the incorporation of histone variants bring about changes in chromatin structure that in turn alter gene expression. Interest in non-allelic histone variants has been renewed, in part because of recent work on H3 (and other) histone variants. However, only in mammals do three non centromeric H3 variants (H3.1, H3.2, and H3.3) exist. Here, we show that mammalian cell lines can be separated into two different groups based on their expression of H3.1, H3.2, and H3.3 at both mRNA and protein levels. Additionally, the ratio of these variants changes slightly during neuronal differentiation of murine ES cells. This difference in H3 variant expression between cell lines could not be explained by changes in growth rate, cell cycle stages, or chromosomal ploidy, but rather suggests other possibilities, such as changes in H3 variant incorporation during differentiation and tissue- or species-specific H3 variant expression. Moreover, quantitative mass spectrometry analysis of human H3.1, H3.2, and H3.3 showed modification differences between these three H3 variants, suggesting that they may have different biological functions. Specifically, H3.3 contains marks associated with transcriptionally active chromatin, whereas H3.2, in contrast, contains mostly silencing modifications that have been associated with facultative heterochromatin. Interestingly, H3.1 is enriched in both active and repressive marks, although the latter marks are different from those observed in H3.2. Although the biological significance as to why mammalian cells differentially employ three highly similar H3 variants remains unclear, our results underscore potential functional differences between them and reinforce the general view that H3.1 and H3.2 in mammalian cells should not be treated as equivalent proteins. PMID- 16267051 TI - Cholesterol-mediated neurite outgrowth is differently regulated between cortical and hippocampal neurons. AB - The acquisition of neuronal type-specific morphogenesis is a central feature of neuronal differentiation and has important consequences for region-specific nervous system functions. Here, we report that the cell type-specific cholesterol profile determines the differential modulation of axon and dendrite outgrowths in hippocampal and cerebral cortical neurons in culture. The extent of axon and dendrite outgrowths is greater and the polarity formation occurs earlier in cortical neurons than in hippocampal neurons. The cholesterol concentrations in total homogenate and the lipid rafts from hippocampal neurons are significantly higher than those from cortical neurons. Cholesterol depletion by beta cyclodextrin markedly enhanced the neurite outgrowth and accelerated the establishment of neuronal polarity in hippocampal neurons, which were similarly observed in nontreated cortical neurons, whereas cholesterol loading had no effects. In contrast, both depletion and loading of cholesterol decreased the neurite outgrowths in cortical neurons. The stimulation of neurite outgrowth and polarity formation induced by cholesterol depletion was accompanied by an enhanced localization of Fyn, a Src kinase, in the lipid rafts of hippocampal neurons. A concomitant treatment with beta-cyclodextrin and a Src family kinase inhibitor, PP2, specifically blocked axon outgrowth but not dendrite outgrowth (both of which were enhanced by beta-cyclodextrin) in hippocampal neurons, suggesting that axon outgrowth modulated by cholesterol is induced in a Fyn dependent manner. These results suggest that cellular cholesterol modulates axon and dendrite outgrowths and neuronal polarization under culture conditions and also that the difference in cholesterol profile between hippocampal and cortical neurons underlies the difference in neurite outgrowth between these two types of neurons. PMID- 16267052 TI - Obese yeast: triglyceride lipolysis is functionally conserved from mammals to yeast. AB - Storage and degradation of triglycerides are essential processes to ensure energy homeostasis and availability of precursors for membrane lipid synthesis. Recent evidence suggests that an emerging class of enzymes containing a conserved patatin domain are centrally important players in lipid degradation. Here we describe the identification and characterization of a major triglyceride lipase of the adipose triglyceride lipase/Brummer family, Tgl4, in the yeast Saccharomyces cerevisiae. Elimination of Tgl4 in a tgl3 background led to fat yeast, rendering growing cells unable to degrade triglycerides. Tgl4 and Tgl3 lipases localized to lipid droplets, independent of each other. Serine 315 in the GXSXG lipase active site consensus sequence of the patatin domain of Tgl4 is essential for catalytic activity. Mouse adipose triglyceride lipase (which also contains a patatin domain but is otherwise highly divergent in primary structure from any yeast protein) localized to lipid droplets when expressed in yeast, and significantly restored triglyceride breakdown in tgl4 mutants in vivo. Our data identify yeast Tgl4 as a functional ortholog of mammalian adipose triglyceride lipase. PMID- 16267053 TI - An aromatic hydroxylation reaction catalyzed by a two-component FMN-dependent Monooxygenase. The ActVA-ActVB system from Streptomyces coelicolor. AB - The ActVA-ActVB system from Streptomyces coelicolor isatwo-component flavin dependent monooxygenase that belongs to an emerging class of enzymes involved in various oxidation reactions in microorganisms. The ActVB component is a NADH:flavin oxidoreductase that provides a reduced FMN to the second component, ActVA the proper monooxygenase. In this work, we demonstrate that the ActVA-ActVB system catalyzes the aromatic monohydroxylation of dihydrokalafungin by molecular oxygen. In the presence of reduced FMN and molecular oxygen, the ActVA active site accommodates and stabilizes an electrophilic flavin FMN-OOH hydroperoxide intermediate species as the oxidant. Surprisingly, we demonstrate that the quinone form of dihydrokalafungin is not oxidized by the ActVA-ActVB system, whereas the corresponding hydroquinone is an excellent substrate. The enantiomer of dihydrokalafungin, nanaomycin A, as well as the enantiomer of kalafungin, nanaomycin D, are also substrates in their hydroquinone forms. The previously postulated product of the ActVA-ActVB system, the antibiotic actinorhodin, was not found to be formed during the oxidation reaction. PMID- 16267054 TI - Focally elevated creatine detected in amyloid precursor protein (APP) transgenic mice and Alzheimer disease brain tissue. AB - The creatine/phosphocreatine system, regulated by creatine kinase, plays an important role in maintaining energy balance in the brain. Energy metabolism and the function of creatine kinase are known to be affected in Alzheimer diseased brain and in cells exposed to the beta-amyloid peptide. We used infrared microspectroscopy to examine hippocampal, cortical, and caudal tissue from 21-89 week-old transgenic mice expressing doubly mutant (K670N/M671L and V717F) amyloid precursor protein and displaying robust pathology from an early age. Microcrystalline deposits of creatine, suggestive of perturbed energetic status, were detected by infrared microspectroscopy in all animals with advanced plaque pathology. Relatively large creatine deposits were also found in hippocampal sections from post-mortem Alzheimer diseased human brain, compared with hippocampus from non-demented brain. We therefore speculate that this molecule is a marker of the disease process. PMID- 16267056 TI - Seven-transmembrane receptor signaling through beta-arrestin. AB - Cell surface receptors are important communicators of external stimuli to the cell interior where they lead to initiation of various signaling pathways and cellular responses. The largest receptor family is the seven-transmembrane receptor (7TMR) family, with approximately 1000 coding genes in the human genome. When 7TMRs are stimulated with agonists, they activate heterotrimeric guanine nucleotide-binding proteins (G proteins), leading to the production of signaling second messengers, such as adenosine 3',5'-monophosphate, inositol phosphates, and others. Activated receptors are rapidly phosphorylated on serine and threonine residues by specialized enzymes called G protein-coupled receptor kinases. Phosphorylated receptors bind the multifunctional adaptor proteins beta arrestin1 and beta-arrestin2 with high affinity. Beta-arrestin binding blocks further G protein coupling, leading to "desensitization" of G protein-dependent signaling pathways. For several years, this was considered the sole function of beta-arrestins. However, novel functions of beta-arrestins have been discovered. Beta-arrestins are now designated as important adaptors that link receptors to the clathrin-dependent pathway of internalization. Beta-arrestins bind and direct the activity of several nonreceptor tyrosine kinases in response to 7TMR stimulation. Beta-arrestins also bind and scaffold members of such signaling cascades as the mitogen-activated protein kinases (MAPKs). Beta-arrestins are crucial components in 7TMR signaling leading to cellular responses that include cell survival and chemotaxis. Beta-arrestins act as endocytic adaptors and signal mediators not only for the 7TMRs, but also for several receptor tyrosine kinases. PMID- 16267055 TI - Fgfr4 is required for effective muscle regeneration in vivo. Delineation of a MyoD-Tead2-Fgfr4 transcriptional pathway. AB - Fgfr4 has been shown to be important for appropriate muscle development in chick limb buds; however, Fgfr4 null mice show no phenotype. Here, we show that staged induction of muscle regeneration in Fgfr4 null mice becomes highly abnormal at the time point when Fgfr4 is normally expressed. By 7 days of regeneration, differentiation of myotubes became poorly coordinated and delayed by both histology and embryonic myosin heavy chain staining. By 14 days much of the muscle was replaced by fat and calcifications. To begin to dissect the molecular pathways involving Fgfr4, we queried the promoter sequences for transcriptional factor binding sites and tested candidate regulators in a 27-time point regeneration series. The Fgfr4 promoter region contained a Tead protein binding site (M-CAT 5'-CATTCCT-3'), and Tead2 showed induction during regeneration commensurate with Fgfr4 regulation. Co-transfection of Tead2 and Fgfr4 promoter reporter constructs into C2C12 myotubes showed Tead2 to activate Fgfr4, and mutation of the M-CAT motif in the Fgfr4 promoter abolished these effects. Immunostaining for Tead2 showed timed expression in myotube nuclei consistent with the mRNA data. Query of the expression timing and genomic sequences of Tead2 suggested direct regulation by MyoD, and consistent with this, MyoD directly bound to two strong E-boxes in the first intron of Tead2 by chromatin immunoprecipitation assay. Moreover, co-transfection of MyoD and Tead2 intron reporter constructs into 10T1/2 cells activated reporter activity in a dose dependent manner. This activation was greatly reduced when the two E-boxes were mutated. Our data suggest a novel MyoD-Tead2-Fgfr4 pathway important for effective muscle regeneration. PMID- 16267058 TI - Recent developments in nonsteroidal antiinflammatory drugs in cats. AB - Pain, particularly chronic pain, is an underestimated ailment in cats. Veterinarians tend to under-diagnose and under-treat pain in this aloof and stoic species. Until recently, there was only one analgesic (i.e., butorphanol) approved in the United States for use in cats; but many analgesics, particularly opioids, have been used extra-label for this purpose. Nonsteroidal antiinflammatory drugs (NSAIDs) have been used sparingly in cats because of safety concerns, which are less of an issue with the newer agents. Meloxicam is the only NSAID labeled for use in cats in the United States, but other agents are available in this country and are labeled for use in cats in other countries. PMID- 16267057 TI - G protein signaling in the regulation of Arabidopsis seed germination. AB - Seed germination and early seedling growth are regulated by an intricate network of hormonal signaling pathways. The plant hormones gibberellic acid (GA) and brassinosteroids (BRs) are important positive regulators of these processes, whereas the hormone abscisic acid (ABA) is a potent negative regulator. In Arabidopsis, analysis of mutant plants has implicated heterotrimeric guanine nucleotide-binding protein (G protein) components in regulation of seed germination by all three of these hormones. PMID- 16267059 TI - Nutritional and herbal therapies in the treatment of heart disease in cats and dogs. AB - Nutritional supplements such as L-carnitine and taurine have been found to be beneficial in dogs and cats with certain cardiac diseases. However, not all animals with cardiac disease respond to nutritional supplementation, which means that further work must be done to identify causes of cardiac disease. Herbal therapies have been used in dogs and cats based on information available from their use in humans. This paper reviews the possible benefits and side effects of L-carnitine, taurine, and herbal supplements. PMID- 16267060 TI - Use of compounded adrenocorticotropic hormone (ACTH) for adrenal function testing in dogs. AB - Serum cortisol concentrations were measured in five healthy dogs in response to five adrenocorticotropic hormone (ACTH) preparations. Cortisol concentrations were similar at time 0 (pre-ACTH) and at 30 and 60 minutes after injection of all forms of ACTH. However, at 90 and 120 minutes post-ACTH, serum cortisol concentrations were significantly lower following injection of two compounded forms of ACTH. The data showed that injection of four compounded forms of ACTH caused elevations in serum cortisol concentrations of a similar magnitude as cosyntropin in samples collected 60 minutes after administration; but concentrations at later times varied, depending on the type of ACTH used. PMID- 16267061 TI - Fungal urinary tract infections in the dog and cat: a retrospective study (2001 2004). AB - Thirty-five animals (23 dogs, 12 cats) with fungal urinary tract infections (UTIs) were retrospectively studied. Dysuria, hematuria, increased frequency of micturition, anorexia, depression, and pyrexia were the most common clinical signs noted. Seven species of fungi were identified in the affected animals. Candida albicans was the most common isolate. Most animals diagnosed with fungal UTI also had other concurrent urinary tract or medical problems. Lower urinary tract diseases, diabetes mellitus, neoplasia, and renal failure were the most common concurrent or preceding diseases identified. Resolution of fungal UTI occurred in 12 animals that received specific antifungal treatment. PMID- 16267062 TI - Toxicity and efficacy of cisplatin and doxorubicin combination chemotherapy for the treatment of canine osteosarcoma. AB - Thirty-five dogs with appendicular osteosarcoma underwent amputation and chemotherapy with cisplatin and doxorubicin every 21 days for up to four cycles. Sixteen dogs completed all four cycles. Two dogs had therapy discontinued because of metastases. The remaining 17 dogs experienced toxicities necessitating protocol alteration or discontinuation. The median survival time of 300 days was not improved over previously reported single-agent protocols, but the 10 dogs that survived to a year lived a median of 510 days. PMID- 16267063 TI - Clinical efficacy of selamectin in the treatment of naturally acquired infection of sucking lice (Linognathus setosus) in dogs. AB - A clinical study was performed in 21 dogs to evaluate the efficacy of selamectin for the treatment of naturally acquired infection of sucking lice (Linognathus setosus [L.setosus]) in dogs. Each dog was randomly assigned to one of two treatment groups. One group was treated with selamectin applied topically at a mean dosage of 7.9 mg/kg. The other group was treated with permethrin applied topically at a mean dosage of 85.7 mg/kg. At day 42 posttreatment, all animals remaining in the study (10 treated with selamectin and six with permethrin) were clear of lice. In both groups, the reduction in lice counts from pretreatment values to day 42 was statistically significant at P< or =0.0001. Selamectin applied topically appeared to be effective against L. setosus infection in dogs. PMID- 16267064 TI - Tibial plateau leveling osteotomy and cranial closing wedge ostectomy in a cat with cranial cruciate ligament rupture. AB - A 5-year-old domestic shorthair cat was presented for a nonweight-bearing left pelvic-limb lameness. Orthopedic and radiographic examinations revealed a cranial cruciate ligament rupture and deformity of the proximal tibial metaphysis. The deformity of the proximal tibia resulted in an exaggerated tibial plateau angle of approximately 75 degrees. Surgical correction was performed with a combination of tibial plateau leveling osteotomy and cranial closing wedge ostectomy. The procedures resulted in an excellent clinical outcome with immediate return to weight bearing in the operated limb and progressive improvement in function over the ensuing 4 months. PMID- 16267065 TI - Subdural hematoma of the brainstem in a dog: magnetic resonance findings and treatment. AB - An 8-year-old, spayed female Dalmatian with a history of seizures was evaluated for cervical pain and bilateral scleral hemorrhages. Diagnostic evaluations revealed a mass displacing the ventral brainstem on magnetic resonance imaging (MRI). The mass was surgically removed and histologically confirmed to be a hematoma. The dog's neurological signs resolved completely after surgery. Although extradural, subdural, subarachnoid, and intraparenchymal hemorrhages have been reported in dogs and cats, this is the first known report of a subdural hematoma of the ventral brainstem in a dog. On the basis of the history and the appearance of the subdural hematoma on MRI, a traumatic event during the seizure episodes was considered the most likely cause of the subdural hematoma in this case. PMID- 16267066 TI - Quadricuspid aortic valve and associated abnormalities in a dog. AB - An 11-month-old, female Scottish terrier was presented with a history of a heart murmur. The electrocardiogram showed signs of left ventricular enlargement, and radiography confirmed generalized cardiomegaly. Echocardiography revealed four equally sized aortic valve cusps. A ventricular septal defect, with systolic left to-right shunting, and aortic regurgitation into both ventricles were also present. The dog was free of clinical signs 1 year after diagnosis. PMID- 16267067 TI - Cardiac toxicity from phenylpropanolamine overdose in a dog. AB - A 5-year-old, 29-kg, female Labrador retriever developed tachypnea, tachycardia, and ataxia following ingestion of approximately 48 mg/kg of phenylpropanolamine. Initial diagnostic tests showed multiform ventricular tachycardia, left ventricular dilatation with a focal dyskinetic region in the dorsal interventricular septum, and elevations in creatinine kinase and cardiac troponin I. All abnormalities resolved within 6 months. The transient electrocardiographic, echocardiographic, and biochemical abnormalities were consistent with myocardial necrosis from infarction or direct catecholamine induced myocardial toxicity. PMID- 16267068 TI - Heart failure improvement from a supplement containing copper. PMID- 16267069 TI - Role of dendritic cells in specific atherosclerosis types. PMID- 16267070 TI - Increased risk of sudden and non-sudden cardiovascular death in patients with atrial fibrillation/flutter following acute myocardial infarction. AB - AIMS: Atrial fibrillation (AF) is a common complication in patients with acute myocardial infarction and is associated with an increase in the risk of death. The excess mortality associated with AF complicating acute myocardial infarction has not been studied in detail. Observations indicate that AF facilitates induction of ventricular arrhythmias, which may increase the risk of sudden cardiovascular death (SCD). A close examination of the mode of death could potentially provide useful knowledge to guide further investigations and treatments. METHODS AND RESULTS: We analysed the relation between AF/atrial flutter (AFL) and modes of death in 5983 consecutive patients discharged alive after an acute myocardial infarction screened in the TRAndolapril Cardiac Evaluation registry. This cohort of patients with an enzyme-verified acute myocardial infarction was admitted to 27 centres in 1990-92. Survival status was obtained 2 years after screening of the last patient. An independent endpoint committee assessed the modes of death. Left ventricular ejection fraction was determined in all the screened patients and information about presence or absence of AF/AFL was prospectively collected. Sustained or paroxysmal AF/AFL was observed in 1149 patients (19%) during hospitalization. During follow-up, 1659 patients (34%) died: 482 (50%) patients with AF/AFL and 1177 (30%) patients without AF/AFL, P<0.001. SCD occurred in 536, non-SCD occurred in 725, and 398 died of non-cardiovascular causes (includes 142 unclassifiable cases). The adjusted risk ratio of AF/AFL for total mortality was 1.33 (95% CI: 1.19-1.49; P<0.0001) and the risk ratio for SCD was 1.31 (95% CI: 1.07-1.60; P<0.009). The adjusted risk ratio of AF/AFL for non-SCD was 1.43 (95% CI: 1.21-1.70; P<0.0001). CONCLUSION: The excess mortality observed in patients with AF/AFL following acute myocardial infarction is due to a significant increase in both SCD and non-SCD. PMID- 16267071 TI - Mobilization of CD34(+), CD117(+), CXCR4(+), c-met(+) stem cells is correlated with left ventricular ejection fraction and plasma NT-proBNP levels in patients with acute myocardial infarction. AB - AIMS: The aim of the study was to assess the correlation between the number of CD34(+), CD117(+), c-met(+), CXCR4(+) stem cells mobilized into peripheral blood, left ventricular ejection fraction (LVEF), NT-proBNP levels, and myocardial necrosis markers in patients with acute myocardial infarction (AMI). METHODS AND RESULTS: 43 patients with STEMI were enrolled. Stem cells number was measured using flow-cytometer and concentrations of NT-proBNP, SDF-1, G-CSF, VEGF, IL-6, and HGF were measured using ELISA kits. The number of stem cells mobilized early (<12 h) in AMI was significantly, positively correlated with LVEF: r=0.49 (P=0.0012) for CD34(+) cells, r=0.48 (P=0.0018) for CXCR4(+) cells, r=0.45 (P=0.0043) for CD117(+) cells, and r=0.41 (P=0.01) for c-met(+) cells and negatively correlated with NT-proBNP levels on admission r=-0.35 (P=0.024) for CD34(+) cells, r=-0.42 (P=0.007) for CXCR4(+) cells, r=-0.33 (P=0.04). In patients with LVEF 40%. The number of CXCR4(+) cells on admission and after 24 h was negatively correlated with respective cardiac Troponin I levels (r=-0.37; P=0.029 and r= 0.45, P=0.02) and maximum activity of CK-MB (r=-0.37; P=0.021). No significant correlations between levels of haematopoietic cytokines and LVEF were found. CONCLUSION: The mobilization of CD34(+), CD117(+), CXCR4(+), c-met(+) stem cells into peripheral blood early in STEMI is positively correlated with LVEF and negatively correlated with NT-proBNP levels and myocardial necrosis markers. PMID- 16267072 TI - Myocardial perfusion scintigraphy: an important step between clinical assessment and coronary angiography in patients with stable chest pain. PMID- 16267073 TI - Ranolazine is associated with cardiovascular and metabolic improvement: a win-win for patients with diabetes. PMID- 16267074 TI - Obstructive sleep apnoea-hypoapnoea syndrome reversibly depresses cardiac response to exercise. AB - AIMS: To evaluate cardiac response to exercise in middle-aged normotensive obstructive sleep apnoea-hypoapnoea syndrome (OSAHS) adults with normal resting left ventricular systolic function and to test the hypothesis that nasal continuous positive airway pressure (CPAP) therapy might improve cardiac performance during exercise. METHODS AND RESULTS: We performed a prospective, randomized, double-blind, placebo-controlled, cross-over clinical trial including 31 consecutive newly diagnosed OSAHS patients and 15 healthy subjects. Cardiopulmonary exercise testing with cardiac output measurement, blood pressure (BP) recordings, and urinary excretion of catecholamine levels were obtained at baseline and after 3 months on both effective and sham CPAP. OSAHS subjects had higher systolic and mean nocturnal BP and higher nocturnal levels of catecholamines. In contrast, they had lower increments in cardiac output (Qt) and in stroke volume (SV) in response to exercise than control subjects. CPAP therapy was associated with highly significant improvements in all the indices of left ventricular systolic performance response during exercise, whereas with sham CPAP, all of them remained unchanged. CONCLUSION: OSAHS patients with normal resting left ventricular systolic function and no hypertension had a worse cardiac response to exercise than healthy subjects. In these patients, 3 months of CPAP improved both Qt and SV responses to exercise. PMID- 16267075 TI - Flow-mediated changes in pulse wave velocity: a new clinical measure of endothelial function. AB - AIMS: To test whether measuring hyperaemic changes in pulse wave velocity (PWV) could be used as a new method of assessing endothelial function for use in clinical practice. METHODS AND RESULTS: Flow-mediated changes in vascular tone may be used to assess endothelial function and may be induced by distal hyperaemia, while endothelium-mediated changes in vascular tone can influence PWV. These three known principles were combined to provide and test a novel method of measuring endothelial function by the acute effects of distal hyperaemia on upper and lower limb PWV (measured by a recently developed method). Flow-mediated changes in upper and lower limb PWV were compared in 17 healthy subjects and seven patients with stable chronic heart failure (CHF), as a condition where endothelial function is impaired but endothelium-independent dilator responses are retained. Corroborative measurements of PWV and brachial artery diameter responses to endothelium-dependent and -independent pharmacological stimuli were performed in a further eight healthy subjects. Flow mediated reduction of PWV (by 14% with no change in blood pressure) was found in normal subjects but was almost abolished in patients with CHF. PWV responses appear to be inversely related to and relatively greater than brachial artery diameter responses. CONCLUSION: The method may offer potential advantages of practical use and sensitivity over conduit artery diameter responses to measure endothelial dysfunction. PMID- 16267076 TI - What is the most cost-effective strategy to screen for left ventricular systolic dysfunction: natriuretic peptides, the electrocardiogram, hand-held echocardiography, traditional echocardiography, or their combination? AB - AIMS: To assess the screening characteristics and cost-effectiveness of screening for left ventricular systolic dysfunction (LVSD) in community subjects. METHODS AND RESULTS: A total of 1392 members of the general public and 928 higher risk subjects were randomly selected from seven community practices. Attending subjects underwent an ECG, N-terminal pro-brain natriuretic peptide (NTproBNP) serum levels, and traditional echocardiography (TE). A total of 533 consecutive subjects underwent hand-held echocardiography (HE). The screening characteristics and cost-effectiveness (cost per case of LVSD diagnosed) of eight strategies to predict LVSD (LVSD <45% on TE) were compared. A total of 1205 subjects attended. Ninety six per cent of subjects with LVSD in the general population had identifiable risk factors. All screening strategies gave excellent negative predictive value. Screening high-risk subjects was most cost-effective, screening low-risk subjects least cost-effective. TE screening was the least cost-effective strategy. NTproBNP screening gave similar cost savings to ECG screening; HE screening greater cost-savings, and HE screening following NTproBNP or ECG pre screening the greatest cost-savings, costing approximately 650 Euros per case of LVSD diagnosed in high-risk subjects (63% cost-savings vs.TE). CONCLUSION: Thus several different modalities allow cost-effective community-based screening for LVSD, especially in high-risk subjects. Such programmes would be cost-effective and miss few cases of LVSD in the community. PMID- 16267077 TI - Transitional care facility for elderly people in hospital awaiting a long term care bed: randomised controlled trial. AB - OBJECTIVE: To assess the effectiveness of moving patients who are waiting in hospital for a long term care bed to an off-site transitional care facility. DESIGN: Randomised controlled trial. SETTING: Three public hospitals in Southern Adelaide. PARTICIPANTS: 320 elderly patients (mean age 83 years) in acute hospital beds (212 randomised to intervention, 108 to control). INTERVENTIONS: A transitional care facility where all patients received a single assessment from a specialist elder care team and appropriate ongoing therapy. MAIN OUTCOME MEASURES: Length of stay in hospital, rates of readmission, deaths, and patient's functional level (modified Barthel index), quality of life (assessment of quality of life), and care needs (residential care scale) at four months. RESULTS: From admission, those in the intervention group stayed a median of 32.5 days (95% confidence interval 29 to 36 days) in hospital. In the control group the median length of stay was 43.5 days (41 to 51 days) (95% confidence interval for difference 6 to 16 days). Patients in the intervention group took a median of 21 days (6 to 27 days) longer to be admitted to permanent care than those in the control group. In both groups few patients went home (14 (7%) in the intervention group v 9 (9%) in the control group). There were no significant differences in death rates (28% v 27%) or rates of transfer back to hospital (28% v 25%). CONCLUSIONS: For frail elderly patients who are awaiting a residential care bed transfer out of hospital to an off-site transitional care unit with focus on aged care "unblocks beds" without adverse effects. PMID- 16267078 TI - Development of a novel home sperm test. AB - BACKGROUND: The majority of men find the production of a semen sample an embarrassing and stressful experience. Consequently, the availability of an over the-counter home sperm test, which would reliably and accurately allow the patient to obtain an assessment of fertility potential at their convenience, would be a major benefit. Our objective was to develop and evaluate a home sperm test that provides a visual estimate of the concentration of progressively motile sperm in a semen sample. METHODS: Three particular challenges are described (i) developing a visualization system; (ii) optimization of the detection limit; and (iii) controlling variation due to changes in ambient temperature. The accuracy of the device was tested against two reference methods: computer-assisted sperm analysis (CASA) and a hyaluronate migration test (HMT). RESULTS: In 129 semen samples, where both reference methods agreed (positive or negative), the accuracy of the device was 95%. The observed likelihood ratio of 8.8 indicated that a sample showing a red line in the device was over eight times more likely to have a positive (normal) result in CASA and HMT than a sample without a red line. CONCLUSIONS: The final device provides a visual estimate of the concentration of progressively motile sperm in a semen sample using a test that is completed within approximately 1 h of production of the sample and can be used by the man in the comfort of his own home. PMID- 16267079 TI - A method for predicting disease subtypes in presence of misclassification among training samples using gene expression: application to human breast cancer. AB - MOTIVATION: An accurate diagnostic and prediction will not be achieved unless the disease subtype status for every training sample used in the supervised learning step is accurately known. Such an assumption requires the existence of a perfect tool for disease diagnostic and classification, which is seldom available in the majority of the cases. Thus, the supervised learning step has to be conducted with a statistical model that contemplates and handles potential mislabeling in the input data. RESULTS: A procedure for handling potential mislabeling among training samples in the prediction of disease subtypes using gene expression data was proposed. A real data-based simulation study about the estrogen receptor status (ER+/ER-) of breast cancer patients was conducted. The results demonstrated that when 1-4 training samples (N = 30) were artificially mislabeled, the proposed method was able not only in correcting the ER status of mislabeled training samples but also more importantly in predicting the ER status of validation samples as well as using 'true' training data. PMID- 16267081 TI - GenoMiner: a tool for genome-wide search of coding and non-coding conserved sequence tags. AB - GenoMiner is a software tool that searches for regions of similarity between user submitted genome or transcript sequences and user-specified whole genome assemblies. The program then identifies conserved sequence tags (CSTs) in these homologous regions and provides a prediction of their coding or non-coding nature. The analysis is carried out through three steps: (1) definition of sequence regions homologous to the query sequence in the selected target genomes by a fast BLAT alignment; (2) identification of CSTs by a more sensitive BLAST like alignment between the query and the homologous regions in the target genomes and (3) assessment of the coding or non-coding nature of detected CSTs through the computation of a suitable coding potential score. GenoMiner allows the user to search the query sequence against a number of vertebrate genome assemblies in a single run providing a user-friendly graphical output. PMID- 16267080 TI - Sequence features of DNA binding sites reveal structural class of associated transcription factor. AB - MOTIVATION: A key goal in molecular biology is to understand the mechanisms by which a cell regulates the transcription of its genes. One important aspect of this transcriptional regulation is the binding of transcription factors (TFs) to their specific cis-regulatory counterparts on the DNA. TFs recognize and bind their DNA counterparts according to the structure of their DNA-binding domains (e.g. zinc finger, leucine zipper, homeodomain). The structure of these domains can be used as a basis for grouping TFs into classes. Although the structure of DNA-binding domains varies widely across TFs generally, the TFs within a particular class bind to DNA in a similar fashion, suggesting the existence of class-specific features in the DNA sequences bound by each class of TFs. RESULTS: In this paper, we apply a sparse Bayesian learning algorithm to identify a small set of class-specific features in the DNA sequences bound by different classes of TFs; the algorithm simultaneously learns a true multi-class classifier that uses these features to predict the DNA-binding domain of the TF that recognizes a particular set of DNA sequences. We train our algorithm on the six largest classes in TRANSFAC, comprising a total of 587 TFs. We learn a six-class classifier for this training set that achieves 87% leave-one-out cross-validation accuracy. We also identify features within cis-regulatory sequences that are highly specific to each class of TF, which has significant implications for how TF binding sites should be modeled for the purpose of motif discovery. PMID- 16267082 TI - libSRES: a C library for stochastic ranking evolution strategy for parameter estimation. AB - SUMMARY: Estimation of kinetic parameters in a biochemical pathway or network represents a common problem in systems studies of biological processes. We have implemented a C library, named libSRES, to facilitate a fast implementation of computer software for study of non-linear biochemical pathways. This library implements a (mu, lambda)-ES evolutionary optimization algorithm that uses stochastic ranking as the constraint handling technique. Considering the amount of computing time it might require to solve a parameter-estimation problem, an MPI version of libSRES is provided for parallel implementation, as well as a simple user interface. libSRES is freely available and could be used directly in any C program as a library function. We have extensively tested the performance of libSRES on various pathway parameter-estimation problems and found its performance to be satisfactory. AVAILABILITY: The source code (in C) is free for academic users at http://csbl.bmb.uga.edu/~jix/science/libSRES/ PMID- 16267083 TI - SSEP-Domain: protein domain prediction by alignment of secondary structure elements and profiles. AB - MOTIVATION: The prediction of protein domains is a crucial task for functional classification, homology-based structure prediction and structural genomics. In this paper, we present the SSEP-Domain protein domain prediction approach, which is based on the application of secondary structure element alignment (SSEA) and profile-profile alignment (PPA) in combination with InterPro pattern searches. SSEA allows rapid screening for potential domain regions while PPA provides us with the necessary specificity for selecting significant hits. The combination with InterPro patterns allows finding domain regions without solved structural templates if sequence family definitions exist. RESULTS: A preliminary version of SSEP-Domain was ranked among the top-performing domain prediction servers in the CASP 6 and CAFASP 4 experiments. Evaluation of the final version shows further improvement over these results together with a significant speed-up. AVAILABILITY: The server is available at http://www.bio.ifi.lmu.de/SSEP/ PMID- 16267084 TI - Using a calibration experiment to assess gene-specific information: full Bayesian and empirical Bayesian models for two-channel microarray data. AB - MOTIVATION: Microarray studies permit to quantify expression levels on a global scale by measuring transcript abundance of thousands of genes simultaneously. A difficulty when analysing expression measures is how to model variability for the whole set of genes. It is usually unrealistic to assume a common variance for each gene. Several approaches to model gene-specific variances are proposed. We take advantage of calibration experiments, in which the probes hybridized on the two channels come from the same population (self-self experiment). In this case it is possible to estimate the gene-specific variance, to be incorporated in comparative experiments on the same tissue, cellular line or species. RESULTS: We present two approaches to introduce prior information on gene-specific variability from a calibration experiment: an empirical Bayes model and a full Bayesian hierarchical model. We apply the methods in the analysis of human lipopolysaccharide-stimulated leukocyte experiments. AVAILABILITY: The calculations are implemented in WinBugs. The codes are available on request from the authors. PMID- 16267085 TI - BioThesaurus: a web-based thesaurus of protein and gene names. AB - BioThesaurus is a web-based system designed to map a comprehensive collection of protein and gene names to protein entries in the UniProt Knowledgebase. Currently covering more than two million proteins, BioThesaurus consists of over 2.8 million names extracted from multiple molecular biological databases according to the database cross-references in iProClass. The BioThesaurus web site allows the retrieval of synonymous names of given protein entries and the identification of protein entries sharing the same names. AVAILABILITY: BioThesaurus is accessible for online searching at http://pir.georgetown.edu/iprolink/biothesaurus PMID- 16267086 TI - Improved spliced alignment from an information theoretic approach. AB - MOTIVATION: mRNA sequences and expressed sequence tags represent some of the most abundant experimental data for identifying genes and alternatively spliced products in metazoans. These transcript sequences are frequently studied by aligning them to a genomic sequence template. For existing programs, error-prone, polymorphic and cross-species data, as well as non-canonical splice sites, still present significant barriers to producing accurate, complete alignments. RESULTS: We took a novel approach to spliced alignment that meaningfully combined information from sequence similarity with that obtained from PSSM splice site models. Scoring systems were chosen to maximize their power of discrimination, and dynamic programming (DP) was employed to guarantee optimal solutions would be found. The resultant program, EXALIN, performed better than other popular tools tested under a wide range of conditions that included detection of micro-exons and human-mouse cross-species comparisons. For improved speed with only a marginal decrease in splice site prediction accuracy, EXALIN could perform limited DP guided by a result from BLASTN. AVAILABILITY: The source code, binaries, scripts, scoring matrices and splice site models for human, mouse, rice and Caenorhabditis elegans utilized in this study are posted at http://blast.wustl.edu/exalin. The software (scripts, source code and binaries) is copyrighted but free for all to use. PMID- 16267088 TI - mILD: a tool for constructing and analyzing matrices of pairwise phylogenetic character incongruence tests. AB - SUMMARY: Pairwise comparisons of disagreement in phylogenetic datasets offer a powerful tool for isolating historical incongruence for closer analysis. Statistically significant phylogenetic character incongruence may reflect important differences in evolutionary history, such as horizontal gene transfer. Such testing can also be used to specify possible combinations of datasets for further phylogenetic analysis. The process of comparing multiple datasets can be very time consuming, and it is sometimes unclear how to combine data partitions given the observed patterns of incongruence. Here we present an application that automates the process of making pairwise comparisons between large numbers of phylogenetic datasets using the Incongruence Length Difference (ILD) test. The application also implements strategies for data combination based on the patterns of incongruence observed in pairwise comparisons. PMID- 16267087 TI - Nexplorer: phylogeny-based exploration of sequence family data. AB - SUMMARY: Nexplorer is a web-based program for interactive browsing and manipulation of character data in NEXUS format, well suited for use with alignments and trees representing families of homologous genes or proteins. Users may upload a sequence family dataset, or choose from one of several thousand already available. Nexplorer provides a flexible means to develop customized views that combine a tree and a data matrix or alignment, to create subsets of data, and to output data files or publication-quality graphics. AVAILABILITY: Web access is from http://www.molevol.org/nexplorer PMID- 16267089 TI - Sequence-based heuristics for faster annotation of non-coding RNA families. AB - MOTIVATION: Non-coding RNAs (ncRNAs) are functional RNA molecules that do not code for proteins. Covariance Models (CMs) are a useful statistical tool to find new members of an ncRNA gene family in a large genome database, using both sequence and, importantly, RNA secondary structure information. Unfortunately, CM searches are extremely slow. Previously, we created rigorous filters, which provably sacrifice none of a CM's accuracy, while making searches significantly faster for virtually all ncRNA families. However, these rigorous filters make searches slower than heuristics could be. RESULTS: In this paper we introduce profile HMM-based heuristic filters. We show that their accuracy is usually superior to heuristics based on BLAST. Moreover, we compared our heuristics with those used in tRNAscan-SE, whose heuristics incorporate a significant amount of work specific to tRNAs, where our heuristics are generic to any ncRNA. Performance was roughly comparable, so we expect that our heuristics provide a high-quality solution that--unlike family-specific solutions--can scale to hundreds of ncRNA families. AVAILABILITY: The source code is available under GNU Public License at the supplementary web site. PMID- 16267090 TI - A genotype calling algorithm for affymetrix SNP arrays. AB - MOTIVATION: A classification algorithm, based on a multi-chip, multi-SNP approach is proposed for Affymetrix SNP arrays. Current procedures for calling genotypes on SNP arrays process all the features associated with one chip and one SNP at a time. Using a large training sample where the genotype labels are known, we develop a supervised learning algorithm to obtain more accurate classification results on new data. The method we propose, RLMM, is based on a robustly fitted, linear model and uses the Mahalanobis distance for classification. The chip-to chip non-biological variance is reduced through normalization. This model-based algorithm captures the similarities across genotype groups and probes, as well as across thousands of SNPs for accurate classification. In this paper, we apply RLMM to Affymetrix 100 K SNP array data, present classification results and compare them with genotype calls obtained from the Affymetrix procedure DM, as well as to the publicly available genotype calls from the HapMap project. PMID- 16267091 TI - Pleiotropic functions of a conserved insect-specific Hox peptide motif. AB - The proteins that regulate developmental processes in animals have generally been well conserved during evolution. A few cases are known where protein activities have functionally evolved. These rare examples raise the issue of how highly conserved regulatory proteins with many roles evolve new functions while maintaining old functions. We have investigated this by analyzing the function of the ;QA' peptide motif of the Hox protein Ultrabithorax (Ubx), a motif that has been conserved throughout insect evolution since its establishment early in the lineage. We precisely deleted the QA motif at the endogenous locus via allelic replacement in Drosophila melanogaster. Although the QA motif was originally characterized as involved in the repression of limb formation, we have found that it is highly pleiotropic. Curiously, deleting the QA motif had strong effects in some tissues while barely affecting others, suggesting that QA function is preferentially required for a subset of Ubx target genes. QA deletion homozygotes had a normal complement of limbs, but, at reduced doses of Ubx and the abdominal A (abd-A) Hox gene, ectopic limb primordia and adult abdominal limbs formed when the QA motif was absent. These results show that redundancy and the additive contributions of activity-regulating peptide motifs play important roles in moderating the phenotypic consequences of Hox protein evolution, and that pleiotropic peptide motifs that contribute quantitatively to several functions are subject to intense purifying selection. PMID- 16267092 TI - Fgf8 expression defines a morphogenetic center required for olfactory neurogenesis and nasal cavity development in the mouse. AB - In vertebrate olfactory epithelium (OE), neurogenesis proceeds continuously, suggesting that endogenous signals support survival and proliferation of stem and progenitor cells. We used a genetic approach to test the hypothesis that Fgf8 plays such a role in developing OE. In young embryos, Fgf8 RNA is expressed in the rim of the invaginating nasal pit (NP), in a small domain of cells that overlaps partially with that of putative OE neural stem cells later in gestation. In mutant mice in which the Fgf8 gene is inactivated in anterior neural structures, FGF-mediated signaling is strongly downregulated in both OE proper and underlying mesenchyme by day 10 of gestation. Mutants survive gestation but die at birth, lacking OE, vomeronasal organ (VNO), nasal cavity, forebrain, lower jaw, eyelids and pinnae. Analysis of mutants indicates that although initial NP formation is grossly normal, cells in the Fgf8-expressing domain undergo high levels of apoptosis, resulting in cessation of nasal cavity invagination and loss of virtually all OE neuronal cell types. These findings demonstrate that Fgf8 is crucial for proper development of the OE, nasal cavity and VNO, as well as maintenance of OE neurogenesis during prenatal development. The data suggest a model in which Fgf8 expression defines an anterior morphogenetic center, which is required not only for the sustenance and continued production of primary olfactory (OE and VNO) neural stem and progenitor cells, but also for proper morphogenesis of the entire nasal cavity. PMID- 16267093 TI - Ligand-dependent de-repression via EcR/USP acts as a gate to coordinate the differentiation of sensory neurons in the Drosophila wing. AB - Loss of function of either the ecdysone receptor (EcR) or Ultraspiracle (USP), the two components of the ecdysone receptor, causes precocious differentiation of the sensory neurons on the wing of Drosophila. We propose that the unliganded receptor complex is repressive and that this repression is relieved as the hormone titers increase at the onset of metamorphosis. The point in development where the receptor complex exerts this repression varies for different groups of sensilla. For the chemosensory organ precursors along the wing margin, the block is at the level of senseless expression and is indirect, via the repressive control of broad expression. Misexpressing broad or senseless can circumvent the repression by the unliganded receptor and leads to precocious differentiation of the sensory neurons. This precocious differentiation results in the misguidance of their axons. The sensory precursors of some of the campaniform sensilla on the third longitudinal vein are born prior to the rise in ecdysone. Their differentiation is also repressed by the unliganded EcR/USP complex but the block occurs after senseless expression but before the precursors undertake their first division. We suggest that in imaginal discs the unliganded EcR/USP complex acts as a ligand-sensitive ;gate' that can be imposed at various points in a developmental pathway, depending on the nature of the cells involved. In this way, the ecdysone signal can function as a developmental timer coordinating development within the imaginal disc. PMID- 16267094 TI - Eph and NMDA receptors control Ca2+/calmodulin-dependent protein kinase II activation during C. elegans oocyte meiotic maturation. AB - Fertilization in the female reproductive tract depends on intercellular signaling mechanisms that coordinate sperm presence with oocyte meiotic progression. To achieve this coordination in Caenorhabditis elegans, sperm release an extracellular signal, the major sperm protein (MSP), to induce oocyte meiotic maturation and ovulation. MSP binds to multiple receptors, including the VAB-1 Eph receptor protein-tyrosine kinase on oocyte and ovarian sheath cell surfaces. Canonical VAB-1 ligands called ephrins negatively regulate oocyte maturation and MPK-1 mitogen-activated protein kinase (MAPK) activation. Here, we show that MSP and VAB-1 regulate the signaling properties of two Ca2+ channels that are encoded by the NMR-1 N-methyl D-aspartate type glutamate receptor subunit and ITR-1 inositol 1,4,5-triphosphate receptor. Ephrin/VAB-1 signaling acts upstream of ITR 1 to inhibit meiotic resumption, while NMR-1 prevents signaling by the UNC-43 Ca2+/calmodulin-dependent protein kinase II (CaMKII). MSP binding to VAB-1 stimulates NMR-1-dependent UNC-43 activation, and UNC-43 acts redundantly in oocytes to promote oocyte maturation and MAPK activation. Our results support a model in which VAB-1 switches from a negative regulator into a redundant positive regulator of oocyte maturation upon binding to MSP. NMR-1 mediates this switch by controlling UNC-43 CaMKII activation at the oocyte cortex. PMID- 16267095 TI - NO-donating aspirin induces phase II enzymes in vitro and in vivo. AB - Modulation of drug metabolizing enzymes, leading to facilitated elimination of carcinogens represents a successful strategy for cancer chemoprevention. Nitric oxide-donating aspirin (NO-ASA) is a promising agent for the prevention of colon and other cancers. We studied the effect of NO-ASA on drug metabolizing enzymes in HT-29 human colon adenocarcinoma and Hepa 1c1c7 mouse liver adenocarcinoma cells and in Min mice treated with NO-ASA for 3 weeks. In these cell lines, NO ASA induced the activity and expression of NAD(P)H:quinone oxireductase (NQO) and glutathione S-transferase (GST). Compared with untreated Min mice, NO-ASA increased in the liver the activity (nmol/min/mg; mean+/-SEM for all) of NQO (85+/-6 versus 128+/-11, P<0.05) and GST (2560+/-233 versus 4254+/-608, P<0.005) and also in the intestine but not in the kidney; the expression of NQO1 and GST P1-1 was also increased. NO-ASA had only a marginal effect on P450 1A1 and P450 2E1, two phase I enzymes. The release of NO from NO-ASA, determined with a selective microelectrode was paralleled by the induction of NQO1 and abrogated by NO scavengers; an exogenous NO donor also induced the expression of NQO1. NO-ASA induced concentration-dependently the translocation of Nrf2 into the nucleus as documented by immunofluorescence and immunoblotting; this paralleled the induction of NQO1 and GST P1-1. Thus NO-ASA induces phase II enzymes, at least in part, through the action of NO that it releases and by modulating the Keap1-Nrf2 pathway; this effect may be part of its mechanism of action against colon and other cancers. PMID- 16267096 TI - Identification of a novel polymorphism Arg290Gln of esophageal cancer related gene 1 (ECRG1) and its related risk to esophageal squamous cell carcinoma. AB - We previously cloned and identified the esophageal cancer related gene 1 (ECRG1), a novel candidate tumor suppressor gene, from human esophageal cells. A single nucleotide polymorphism (Arg290Gln) was identified in the coding region of ECRG1 and might play a role in susceptibility to esophageal squamous cell carcinoma (ESCC). To examine this hypothesis, we analyzed 998 ESCC patients and 1252 controls in a hospital-based, case-control study in a Chinese population for this polymorphism. We observed a statistically significantly increased risk of ESCC associated with the ECRG1 290Arg/Gln and 290Gln/Gln genotypes compared with the 290Arg/Arg [odds ratio (OR)=1.23, 95% confidence interval (CI)=1.03-1.46; P<0.05]. A greater than multiplicative joint effect between the ECRG1 polymorphism and tobacco smoking exposure was also observed (OR=1.95, 95% CI=1.48 2.56; P<0.001). Furthermore, the elevated risk of ESCC associated with the ECRG1 polymorphism was increased consistently with cumulative smoking dose. ORs (95% CI) for 290Arg/Gln and 290Gln/Gln genotypes among non-smokers and smokers who smoked27 pack-years were 1.03 (0.78-1.35), 1.91 (1.36-2.67) and 2.08 (1.48-2.92), respectively (P trend test<0.001). Taken together, our results indicate that the ECRG1 290Gln variant allele might be a genetic susceptibility factor for developing ESCC, especially in the smoking population. PMID- 16267097 TI - Dietary HDAC inhibitors: time to rethink weak ligands in cancer chemoprevention? AB - There is growing interest in the various mechanisms that regulate chromatin remodeling, including modulation of histone deacetylase (HDAC) activities. Competitive HDAC inhibitors disrupt the cell cycle and/or induce apoptosis via de repression of genes such as P21 and BAX, and cancer cells appear to be more sensitive than non-transformed cells to trichostatin A and related HDAC inhibitory compounds. This apparent selectivity of action in cancer cells makes HDAC inhibitors an attractive avenue for drug development. However, in the search for potent HDAC inhibitors with cancer therapeutic potential there has been a tendency to overlook or dismiss weak ligands that could prove effective in cancer prevention, including agents present in the human diet. Recent reports have described butyrate, diallyl disulfide and sulforaphane as HDAC inhibitors, and many other dietary agents will be probably discovered to attenuate HDAC activity. Here we discuss 'pharmacologic' agents that potently de-repress gene expression (e.g. during therapeutic intervention) versus dietary HDAC inhibitors that, as weak ligands, might subtly regulate the expression of genes involved in cell growth and apoptosis. An important question is the extent to which dietary HDAC inhibitors, and other dietary agents that affect gene expression via chromatin remodeling, modulate the expression of genes such as P21 and BAX so that cells can respond most effectively to external stimuli and toxic insults. PMID- 16267100 TI - Work ability of workers in western China: reference data. AB - BACKGROUND: The Work Ability Index (WAI) is a validated and widely used research tool. Reference data for Chinese workers by age, gender and work content are poorly documented or lacking. OBJECTIVE: To provide reference data for work ability among workers in western China. METHODS: A random sample of 10 218 workers (including manual, professional, clerical and semi-skilled workers) in western China, aged 16-69 years, was taken from several studies and the WAI questionnaire was administered. RESULTS: All the WAI scores were distributed continuously and nearly normally. The WAI for female workers was significantly higher than for males (P < 0.01) and the 'poor' WAI category only accounted for 3% of females, against 6% of males. WAI scores declined with age and changed variably among workers according to work content and age. Mean WAI scores of manual workers declined rapidly beyond the age of 35 years, and beyond age 45 years for professional and clerical workers. WAI scores were distributed differently according to work content and age group (P < 0.01). WAI categories differed by work content and age group both for male and female workers. CONCLUSION: WAI is validated in Chinese occupational practices. Some reference data are different from Finnish data. This study provides gender, age and work content-specific WAI reference values that will help enable comparison and intervention evaluation in further studies. PMID- 16267098 TI - A front-end desaturase from Chlamydomonas reinhardtii produces pinolenic and coniferonic acids by omega13 desaturation in methylotrophic yeast and tobacco. AB - Pinolenic acid (PA; 18:3Delta(5,9,12)) and coniferonic acid (CA; 18:4Delta(5,9,12,15)) are Delta(5)-unsaturated bis-methylene-interrupted fatty acids (Delta(5)-UBIFAs) commonly found in pine seed oil. They are assumed to be synthesized from linoleic acid (LA; 18:2Delta(9,12)) and alpha-linolenic acid (ALA; 18:3Delta(9,12,15)), respectively, by Delta(5)-desaturation. A unicellular green microalga Chlamydomonas reinhardtii also accumulates PA and CA in a betain lipid. The expressed sequence tag (EST) resource of C. reinhardtii led to the isolation of a cDNA clone that encoded a putative fatty acid desaturase named as CrDES containing a cytochrome b5 domain at the N-terminus. When the coding sequence was expressed heterologously in the methylotrophic yeast Pichia pastoris, PA and CA were newly detected and comparable amounts of LA and ALA were reduced, demonstrating that CrDES has Delta(5)-desaturase activity for both LA and ALA. CrDES expressed in the yeast showed Delta(5)-desaturase activity on 18:1Delta(9) but not 18:1Delta(11). Unexpectedly, CrDES also showed Delta(7) desaturase activity on 20:2Delta(11,14) and 20:3Delta(11,14,17) to produce 20:3Delta(7,11,14) and 20:4Delta(7,11,14,17), respectively. Since both the Delta(5) bond in C18 and the Delta(7) bond in C20 fatty acids are 'omega13' double bonds, these results indicate that CrDES has omega13 desaturase activity for omega9 unsaturated C18/C20 fatty acids, in contrast to the previously reported front-end desaturases. In order to evaluate the activity of CrDES in higher plants, transgenic tobacco plants expressing CrDES were created. PA and CA accumulated in the leaves of transgenic plants. The highest combined yield of PA and CA was 44.7% of total fatty acids, suggesting that PA and CA can be produced in higher plants on a large scale. PMID- 16267099 TI - Apoplastic glycosidases active against xyloglucan oligosaccharides of Arabidopsis thaliana. AB - All four glycanases necessary for the degradation of xyloglucan oligosaccharides (alpha-fucosidase, alpha-xylosidase, beta-galactosidase and beta-glucosidase) were found in the apoplastic fluid of Arabidopsis thaliana. These activities acted cooperatively on xyloglucan oligosaccharides (XLFG), leading to the sequential formation of XXFG, XXLG, XXXG, GXXG and XXG, as identified by matrix assisted laser desorption ionization time of flight (MALDI-TOF). AtFXG1 (At1g67830) and AtXYL1 (At1g68560) had been previously identified as the Arabidopsis genes coding for alpha-fucosidase and alpha-xylosidase, respectively. As for the genes coding for beta-galactosidase activity, we identified in phylogenetic trees 12 candidates from family 35 of glycoside hydrolases. Similarly, four genes from family 3 were selected as possible beta-glucosidases active on xyloglucan. The expression level of all the selected genes was studied in different plant regions (young and mature rosette leaves, apical and basal region of the inflorescence stem, roots, flower and siliques) using quantitative real-time reverse transcription-PCR. The expression patterns were very diverse as well as their relationship with growth rates, showing a very complex situation. This could lead to highly varying proportions of the different xyloglucan oligosaccharides in different plant regions and developmental stages. PMID- 16267101 TI - A longitudinal study of the influence of shift work on serum uric acid levels in workers at a telecommunications company. AB - BACKGROUND: Hyperuricemia is a lifestyle-related disease. Although there have been many previous reports about the association of serum uric acid (UA) levels with lifestyle, including eating habits and alcohol intake, there has been no report of a longitudinal study of the relationship between serum UA levels and shift work. AIMS: To clarify the influence of shift work on serum UA levels in Japanese workers. METHODS: This was a 4-year cohort study of 15 871 workers at a telecommunications company. Pooled logistic regression analyses by sex were performed, with job schedule type, age, body mass index (BMI), lifestyle and the results of blood chemistries as covariates. RESULTS: In males, shift work, part time work, BMI, consumption of alcohol (less than twice per week, two to five times per week or more than five times per week) and little preference for vegetables were positively associated with the development of increased serum UA (>or=8 mg/dl in males, >or=6 mg/dl in females). In females, age, BMI and a history of smoking were positively associated with the development of increased serum UA. CONCLUSION: This study revealed that shift work is independently related to increased serum UA in males. PMID- 16267102 TI - Occupational fitness standards for beach lifeguards. Phase 2: the development of an easily administered fitness test. AB - BACKGROUND: No task-based fitness standard currently exists for beach lifeguards (BLGs). AIM: To formulate an easily administered fitness test for BLGs based on the physical demands identified in Phase 1 of the project (previous paper). METHODS: A range of anthropometric and land- and water-based (swimming pool and flume) fitness assessments were administered to 25 male and female volunteer subjects (13 BLGs from the UK). RESULTS: The mean (SD) VO(2max) (l/min) were 3.04 (0.61) for towing a casualty, 2.08 (0.53) for board paddling with a casualty and 2.97 (0.67) for freestyle swimming. A significant correlation (r = -0.82, P < 0.001) was identified between distance paddled in the sea in 3.5 min (established in Phase 1) and pool 400-m front crawl swim time and between towing VO(2max) and deltoid circumference/log(10) 400-m front crawl swim time (r = -0.83, P < 0.001). CONCLUSIONS: The regression identified allows the conclusion that if a BLG can swim 400-m front crawl in a pool in <7.5 min, he/she should be able to paddle 310 m in the sea in <3.5 min. Final recommendations for a fitness test for potential BLGs are presented. PMID- 16267104 TI - Mouse GPR40 heterologously expressed in Xenopus oocytes is activated by short-, medium-, and long-chain fatty acids. AB - Several orphan G protein-coupled receptors, including GPR40, have recently been shown to be responsive to fatty acids. Although previous reports have suggested GPR40 detects medium- and long-chain fatty acids, it has been reported to be unresponsive to short chain fatty acids. In this study, we have heterologously expressed mouse GPR40 in Xenopus laevis oocytes and measured fatty acid-induced increases in intracellular Ca(2+), via two electrode voltage clamp recordings of the endogenous Ca(2+)-activated chloride conductance. Exposure to 500 muM linoleic acid (C18:2), a long-chain fatty acid, stimulated significant currents in mGPR40-injected oocytes (P < 0.01, ANOVA), but not in water-injected control oocytes (not significant, ANOVA). These currents were confirmed as Ca(2+) activated chloride conductances because they were biphasic, sensitive to changes in external pH, and inhibited by DIDS. Similar currents were observed with medium chain fatty acids, such as lauric acid (C12:0) (P < 0.01, ANOVA), and more importantly, with short-chain fatty acids, such as butyric acid (C4:0) (P < 0.01, ANOVA). In contrast, no responses were observed in mGPR40-injected oocytes exposed to either acetic acid (C2:0) or propionic acid (C3:0). Therefore, GPR40 has the capacity to respond to fatty acids with chain lengths of four or greater. This finding has important implications for understanding the structure:function relationship of fatty acid sensors, and potentially for short-chain fatty acid sensing in the gastrointestinal tract. PMID- 16267105 TI - High mobility group box 1 protein interacts with multiple Toll-like receptors. AB - High mobility group box 1 (HMGB1), originally described as a DNA-binding protein, can also be released extracellularly and functions as a late mediator of inflammatory responses. Although recent reports have indicated that the receptor for advanced glycation end products (RAGE) as well as Toll-like receptor (TLR)2 and TLR4 are involved in cellular activation by HMGB1, there has been little evidence of direct association between HMGB1 and these receptors. To examine this issue, we used fluorescence resonance energy transfer (FRET) and immunoprecipitation to directly investigate cell surface interactions of HMGB1 with TLR2, TLR4, and RAGE. FRET images in RAW264.7 macrophages demonstrated association of HMGB1 with TLR2 and TLR4 but not RAGE. Transient transfections into human embryonic kidney-293 cells showed that HMGB1 induced cellular activation and NF-kappaB-dependent transcription through TLR2 or TLR4 but not RAGE. Coimmunoprecipitation also found interaction between HMGB1 and TLR2 as well as TLR4, but not with RAGE. These studies provide the first direct evidence that HMGB1 can interact with both TLR2 and TLR4 and also supply an explanation for the ability of HMGB1 to induce cellular activation and generate inflammatory responses that are similar to those initiated by LPS. PMID- 16267103 TI - TNF-alpha dilates cerebral arteries via NAD(P)H oxidase-dependent Ca2+ spark activation. AB - Expression of TNF-alpha, a pleiotropic cytokine, is elevated during stroke and cerebral ischemia. TNF-alpha regulates arterial diameter, although mechanisms mediating this effect are unclear. In the present study, we tested the hypothesis that TNF-alpha regulates the diameter of resistance-sized ( approximately 150 microm diameter) cerebral arteries by modulating local and global intracellular Ca(2+) signals in smooth muscle cells. Laser-scanning confocal imaging revealed that TNF-alpha increased Ca(2+) spark and Ca(2+) wave frequency but reduced global intracellular Ca(2+) concentration ([Ca(2+)](i)) in smooth muscle cells of intact arteries. TNF-alpha elevated reactive oxygen species (ROS) in smooth muscle cells of intact arteries, and this increase was prevented by apocynin or diphenyleneiodonium (DPI), both of which are NAD(P)H oxidase blockers, but was unaffected by inhibitors of other ROS-generating enzymes. In voltage-clamped (-40 mV) cells, TNF-alpha increased the frequency and amplitude of Ca(2+) spark induced, large-conductance, Ca(2+)-activated K(+) (K(Ca)) channel transients approximately 1.7- and approximately 1.4-fold, respectively. TNF-alpha-induced transient K(Ca) current activation was reversed by apocynin or by Mn(III)tetrakis(1-methyl-4-pyridyl)porphyrin (MnTMPyP), a membrane-permeant antioxidant, and was prevented by intracellular dialysis of catalase. TNF-alpha induced reversible and similar amplitude dilations in either endothelium-intact or endothelium-denuded pressurized (60 mmHg) cerebral arteries. MnTMPyP, thapsigargin, a sarcoplasmic reticulum Ca(2+)-ATPase blocker that inhibits Ca(2+) sparks, and iberiotoxin, a K(Ca) channel blocker, reduced TNF-alpha-induced vasodilations to between 15 and 33% of control. In summary, our data indicate that TNF-alpha activates NAD(P)H oxidase, resulting in an increase in intracellular H(2)O(2) that stimulates Ca(2+) sparks and transient K(Ca) currents, leading to a reduction in global [Ca(2+)](i), and vasodilation. PMID- 16267106 TI - Evidence for Ca(2+)-permeable AMPA receptors in the olfactory bulb. AB - alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors (AMPARs), a subtype of glutamate receptor, contribute to olfactory processing in the olfactory bulb (OB). These ion channels consist of various combinations of the subunits GluR1-GluR4, which bestow certain properties. For example, AMPARs that lack GluR2 are highly permeable to Ca(2+) and generate inwardly rectifying currents. Because increased intracellular Ca(2+) could trigger a host of Ca(2+) dependent odor-encoding processes, we used whole cell recording as well as histological and immunocytochemical (ICC) techniques to investigate whether AMPARs on rat OB neurons flux Ca(2+). Application of 1-naphthylacetyl spermine (NAS), a selective antagonist of Ca(2+)-permeable AMPARs (CP-AMPARs), inhibited AMPAR-mediated currents in subsets of interneurons and principal cells in cultures and slices. The addition of spermine to the electrode yielded inwardly rectifying current-voltage plots in some cells. In OB slices, olfactory nerve stimulation elicited excitatory responses in juxtaglomerular and mitral cells. Bath application of NAS with d,l-2-amino-5-phosphonovaleric acid (AP5) to isolate AMPARs suppressed the amplitudes of these synaptic responses compared with responses obtained using AP5 alone. Co(2+) staining, which involves the kainate stimulated influx of Co(2+) through CP-AMPARs, produced diverse patterns of labeling in cultures and slices as did ICC techniques used with a GluR2-selective antibody. These results suggest that subsets of OB neurons express CP-AMPARs, including functional CP-AMPARs at synapses. Ca(2+) entry into cells via these receptors could influence odor encoding by modulating K(+) channels, N-methyl-d aspartate receptors, and Ca(2+)-binding proteins, or it could facilitate synaptic vesicle fusion. PMID- 16267107 TI - Divergent kinase signaling mediates agonist-induced phosphorylation of phosphatase inhibitory proteins PHI-1 and CPI-17 in vascular smooth muscle cells. AB - Phosphatase holoenzyme inhibitor (PHI)-1 is one of the newest members of the family of protein phosphatase inhibitor proteins. In isolated enzyme systems, several kinases, including PKC and rho kinase (ROCK), have been shown to phosphorylate PHI-1. However, it is largely unknown whether PHI-1 is phosphorylated in response to agonist stimulation in intact cells. We investigated this question in primary cultured rat aortic vascular smooth muscle cells (VSMCs). Using two-dimensional polyacrylamide gel electrophoresis and immunoblot, we found that there are two major PHI-1 spots under resting conditions: a minor spot with an acidic isoelectric point (pI) and a major spot with a more alkaline pI. Interestingly, U-46619, a G protein-coupled receptor agonist, caused a significant increase in the acidic spot, suggesting that it may represent a phosphorylated form of PHI-1. This was confirmed by phosphatase treatment and by a specific phospho-PHI-1 antibody. Furthermore, we found that angiotensin II, thrombin, and U-46619 increased phosphorylated PHI-1 from 9% of total PHI-1 in resting cells to 18%, 18%, and 30%, respectively. We also found that inhibition of ROCK by Y-27632 or H-1152 selectively diminished U-46619 induced CPI-17 phosphorylation, whereas it did not affect PHI-1 phosphorylation. Activation of ROCK by expressing V14RhoA selectively induced CPI-17 phosphorylation without affecting PHI-1 phosphorylation. In contrast, inhibition of PKC by GF-109203X or by PKC downregulation selectively diminished U-46619 induced PHI-1 phosphorylation without significantly affecting U-46619-induced CPI 17 phosphorylation. Activating PKC by PMA induced PHI-1 phosphorylation. Together, our results show for the first time that agonist induces PHI-1 phosphorylation in VSMCs and divergent kinase signaling couples agonist stimulation to PHI-1 and CPI-17 phosphorylation. PMID- 16267108 TI - Sphingosine-1-phosphate activates BKCa channels independently of G protein coupled receptor in human endothelial cells. AB - The effect of sphingosine-1-phosphate (S1P) on large-conductance Ca(2+)-activated K(+) (BK(Ca)) channels was examined in primary cultured human umbilical vein endothelial cells by measuring intracellular Ca(2+) concentration ([Ca(2+)](i)), whole cell membrane currents, and single-channel activity. In nystatin-perforated current-clamped cells, S1P hyperpolarized the membrane and simultaneously increased [Ca(2+)](i). [Ca(2+)](i) and membrane potentials were strongly correlated. In whole cell clamped cells, BK(Ca) currents were activated by increasing [Ca(2+)](i) via cell dialysis with pipette solution, and the activated BK(Ca) currents were further enhanced by S1P. When [Ca(2+)](i) was buffered at 1 microM, the S1P concentration required to evoke half-maximal activation was 403 +/- 13 nM. In inside-out patches, when S1P was included in the bath solution, S1P enhanced BK(Ca) channel activity in a reversible manner and shifted the relationship between Ca(2+) concentration in the bath solution and the mean open probability to the left. In whole cell clamped cells or inside-out patches loaded with guanosine 5'-O-(2-thiodiphosphate) (GDPbetaS; 1 mM) using a patch pipette, GDPbetaS application or pretreatment of cells with pertussis toxin (100 ng/ml) for 15 h did not affect S1P-induced BK(Ca) current and channel activation. These results suggest that S1P enhances BK(Ca) channel activity by increasing Ca(2+) sensitivity. This channel activation hyperpolarizes the membrane and thereby increases Ca(2+) influx through Ca(2+) entry channels. Inasmuch as S1P activates BK(Ca) channels via a mechanism independent of G protein-coupled receptors, S1P may be a component of the intracellular second messenger that is involved in Ca(2+) mobilization in human endothelial cells. PMID- 16267109 TI - MAP kinase and calcium signaling mediate fluid flow-induced human mesenchymal stem cell proliferation. AB - Mechanical signals are important regulators of skeletal homeostasis, and strain induced oscillatory fluid flow is a potent mechanical stimulus. Although the mechanisms by which osteoblasts and osteocytes respond to fluid flow are being elucidated, little is known about the mechanisms by which bone marrow-derived mesenchymal stem cells respond to such stimuli. Here we show that the intracellular signaling cascades activated in human mesenchymal stem cells by fluid flow are similar to those activated in osteoblastic cells. Oscillatory fluid flow inducing shear stresses of 5, 10, and 20 dyn/cm(2) triggered rapid, flow rate-dependent increases in intracellular calcium that pharmacological studies suggest are inositol trisphosphate mediated. The application of fluid flow also induced the phosphorylation of extracellular signal-regulated kinase-1 and -2 as well as the activation of the calcium-sensitive protein phosphatase calcineurin in mesenchymal stem cells. Activation of these signaling pathways combined to induce a robust increase in cellular proliferation. These data suggest that mechanically induced fluid flow regulates not only osteoblastic behavior but also that of mesenchymal precursors, implying that the observed osteogenic response to mechanical loading may be mediated by alterations in the cellular behavior of multiple members of the osteoblast lineage, perhaps by a common signaling pathway. PMID- 16267110 TI - EMG changes in human thenar motor units with force potentiation and fatigue. AB - Few studies have analyzed activity-induced changes in EMG activity in individual human motor units. We studied the changes in human thenar motor unit EMG that accompany the potentiation of twitch force and fatigue of tetanic force. Single motor unit EMG and force were recorded in healthy subjects in response to selective stimulation of their motor axons within the median nerve just above the elbow. Twitches were recorded before and after a series of pulse trains delivered at frequencies that varied between 5 and 100 Hz. This stimulation induced significant increases in EMG amplitude, duration, and area. However, in relative terms, all of these EMG changes were substantially smaller than the potentiation of twitch force. Another 2 min of stimulation (13 pulses at 40 Hz each second) induced additional potentiation of EMG amplitude, duration, and area, but the tetanic force from every unit declined. Thus activity-induced changes in human thenar motor unit EMG do not indicate the alterations in force or vice versa. These data suggest that different processes underlie the changes in EMG and force that occur during human thenar motor unit activity. PMID- 16267112 TI - Temperature modulates taste responsiveness and stimulates gustatory neurons in the rat geniculate ganglion. AB - In humans, temperature influences taste intensity and quality perception, and thermal stimulation itself may elicit taste sensations. However, peripheral coding mechanisms of taste have generally been examined independently of the influence of temperature. In anesthetized rats, we characterized the single-cell responses of geniculate ganglion neurons to 0.5 M sucrose, 0.1 M NaCl, 0.01 M citric acid, and 0.02 M quinine hydrochloride at a steady, baseline temperature (adapted) of 10, 25, and 40 degrees C; gradual cooling and warming (1 degrees C/s change in water temperature >5 s) from an adapted tongue temperature of 25 degrees C; gradual cooling from an adapted temperature of 40 degrees C; and gradual warming from an adapted temperature of 10 degrees C. Hierarchical cluster analysis of the taste responses at 25 degrees C divided 50 neurons into two major categories of narrowly tuned (Sucrose-specialists, NaCl-specialists) and broadly tuned (NaCl-generalists(I), NaCl- generalists(II), Acid-generalists, and QHCl generalists) groups. NaCl specialists were excited by cooling from 25 to 10 degrees C and inhibited by warming from 10 to 25 degrees C. Acid-generalists were excited by cooling from 40 to 25 degrees C but not from 25 to 10 degrees C. In general, the taste responses of broadly tuned neurons decreased systematically to all stimuli with decreasing adapted temperatures. The response selectivity of Sucrose-specialists for sucrose and NaCl-specialists for NaCl was unaffected by adapted temperature. However, Sucrose-specialists were unresponsive to sucrose at 10 degrees C, whereas NaCl-specialists responded equally to NaCl at all adapted temperatures. In conclusion, we have shown that temperature modulates taste responsiveness and is itself a stimulus for activation in specific types of peripheral gustatory neurons. PMID- 16267111 TI - Neonatal cortical ablation disrupts multisensory development in superior colliculus. AB - The ability of cat superior colliculus (SC) neurons to synthesize information from different senses depends on influences from two areas of the cortex: the anterior ectosylvian sulcus (AES) and the rostral lateral suprasylvian sulcus (rLS). Reversibly deactivating the inputs to the SC from either of these areas in normal adults severely compromises this ability and the SC-mediated behaviors that depend on it. In this study, we found that removal of these areas in neonatal animals precluded the normal development of multisensory SC processes. At maturity there was a substantial decrease in the incidence of multisensory neurons, and those multisensory neurons that did develop were highly abnormal. Their cross-modal receptive field register was severely compromised, as was their ability to integrate cross-modal stimuli. Apparently, despite the impressive plasticity of the neonatal brain, it cannot compensate for the early loss of these cortices. Surprisingly, however, neonatal removal of either AES or rLS had comparatively minor consequences on these properties. At maturity multisensory SC neurons were quite common: they developed the characteristic spatial register among their unisensory receptive fields and exhibited normal adult-like multisensory integration. These observations suggest that during early ontogeny, when the multisensory properties of SC neurons are being crafted, AES and rLS may have the ability to compensate for the loss of one another's cortico-collicular influences so that normal multisensory processes can develop in the SC. PMID- 16267113 TI - Coding of stimulus frequency by latency in thalamic networks through the interplay of GABAB-mediated feedback and stimulus shape. AB - A temporal sensory code occurs in posterior medial (POm) thalamus of the rat vibrissa system, where the latency for the spike rate to peak is observed to increase with increasing frequency of stimulation between 2 and 11 Hz. In contrast, the latency of the spike rate in the ventroposterior medial (VPm) thalamus is constant in this frequency range. We consider the hypothesis that two factors are essential for latency coding in the POm. The first is GABAB-mediated feedback inhibition from the reticular thalamic (Rt) nucleus, which provides delayed and prolonged input to thalamic structures. The second is sensory input that leads to an accelerating spike rate in brain stem nuclei. Essential aspects of the experimental observations are replicated by the analytical solution of a rate-based model with a minimal architecture that includes only the POm and Rt nuclei, i.e., an increase in stimulus frequency will increase the level of inhibitory output from Rt thalamus and lead to a longer latency in the activation of POm thalamus. This architecture, however, admits period-doubling at high levels of GABAB-mediated conductance. A full architecture that incorporates the VPm nucleus suppresses period-doubling. A clear match between the experimentally measured spike rates and the numerically calculated rates for the full model occurs when VPm thalamus receives stronger brain stem input and weaker GABAB mediated inhibition than POm thalamus. Our analysis leads to the prediction that the latency code will disappear if GABAB-mediated transmission is blocked in POm thalamus or if the onset of sensory input is too abrupt. We suggest that GABAB mediated inhibition is a substrate of temporal coding in normal brain function. PMID- 16267115 TI - Computational estimation of the distribution of L-type Ca(2+) channels in motoneurons based on variable threshold of activation of persistent inward currents. AB - In the presence of neuromodulators such as serotonin and noradrenaline, motoneurons exhibit persistent inward currents (PICs) that serve to amplify synaptic inputs. A major component of these PICs is mediated by L-type Ca(2+) channels. Estimates based on electrophysiological studies indicate that these channels are located on the dendrites, but immunohistochemical studies of their precise distribution have yielded different results. Our goal was to determine the distribution of these channels using computational methods. A theoretical analysis of the activation of PICs by a somatic current injection in the absence or presence of synaptic activity suggests that L-type Ca(2+) channels may be segregated to discrete hot spots 25-200 microm long and centered 100-400 microm from the soma in the dendritic tree. Compartmental models based on detailed anatomical measurements of the structure of feline neck motoneurons with L-type Ca(2+) channels incorporated in these regions produced plateau potentials resulting from PIC activation. Furthermore, we replicated the experimental observation that the somatic threshold at which PICs were activated was depolarized by tonic activation of inhibitory synapses and hyperpolarized by tonic activation of excitatory synapses. Models with L-type Ca(2+) channels distributed uniformly were unable to replicate the change in somatic threshold of PIC activation. Therefore we conclude that the set of L-type Ca(2+) channels mediating plateau potentials is restricted to discrete regions in the dendritic tree. Furthermore, this distribution leads to the compartmentalization of the dendritic tree of motoneurons into subunits whose sequential activation lead to the graded amplification of synaptic inputs. PMID- 16267114 TI - Intermittent visuomotor processing in the human cerebellum, parietal cortex, and premotor cortex. AB - The cerebellum, parietal cortex, and premotor cortex are integral to visuomotor processing. The parameters of visual information that modulate their role in visuomotor control are less clear. From motor psychophysics, the relation between the frequency of visual feedback and force variability has been identified as nonlinear. Thus we hypothesized that visual feedback frequency will differentially modulate the neural activation in the cerebellum, parietal cortex, and premotor cortex related to visuomotor processing. We used functional magnetic resonance imaging at 3 Tesla to examine visually guided grip force control under frequent and infrequent visual feedback conditions. Control conditions with intermittent visual feedback alone and a control force condition without visual feedback were examined. As expected, force variability was reduced in the frequent compared with the infrequent condition. Three novel findings were identified. First, infrequent (0.4 Hz) visual feedback did not result in visuomotor activation in lateral cerebellum (lobule VI/Crus I), whereas frequent (25 Hz) intermittent visual feedback did. This is in contrast to the anterior intermediate cerebellum (lobule V/VI), which was consistently active across all force conditions compared with rest. Second, confirming previous observations, the parietal and premotor cortices were active during grip force with frequent visual feedback. The novel finding was that the parietal and premotor cortex were also active during grip force with infrequent visual feedback. Third, right inferior parietal lobule, dorsal premotor cortex, and ventral premotor cortex had greater activation in the frequent compared with the infrequent grip force condition. These findings demonstrate that the frequency of visual information reduces motor error and differentially modulates the neural activation related to visuomotor processing in the cerebellum, parietal cortex, and premotor cortex. PMID- 16267116 TI - Processing of kinetic boundaries in macaque V4. AB - We used gratings and shapes defined by relative motion to study selectivity for static kinetic boundaries in macaque V4 neurons. Kinetic gratings were generated by random pixels moving in opposite directions in the neighboring bars, either parallel to the orientation of the boundary (parallel kinetic grating) or perpendicular to the boundary (orthogonal kinetic grating). Neurons were also tested with static, luminance defined gratings to establish cue invariance. In addition, we used eight shapes defined either by relative motion or by luminance contrast, as used previously to test cue invariance in the infero-temporal (IT) cortex. A sizeable fraction (10-20%) of the V4 neurons responded selectively to kinetic patterns. Most neurons selective for kinetic contours had receptive fields (RFs) within the central 10 degrees of the visual field. Neurons selective for the orientation of kinetic gratings were defined as having similar orientation preferences for the two types of kinetic gratings, and the vast majority of these neurons also retained the same orientation preference for luminance defined gratings. Also, kinetic shape selective neurons had similar shape preferences when the shape was defined by relative motion or by luminance contrast, showing a cue-invariant form processing in V4. Although shape selectivity was weaker in V4 than what has been reported in the IT cortex, cue invariance was similar in the two areas, suggesting that invariance for luminance and motion cues of IT originates in V4. The neurons selective for kinetic patterns tended to be clustered within dorsal V4. PMID- 16267118 TI - Two kinematic synergies in voluntary whole-body movements during standing. AB - We used a particular computational approach, the uncontrolled manifold hypothesis, to investigate joint angle covariation patterns during whole-body actions performed by standing persons. We hypothesized that two kinematic synergies accounted for the leg/trunk joint covariation across cycles during a rhythmic whole-body motion to stabilize two performance variables, the trunk orientation in the external space and the horizontal position of the center of mass (COM). Subjects stood on a force plate and performed whole-body rhythmic movements for 45 s under visual feedback on one of the four variables, the position of the center of pressure or the angle in one of the three joints (ankle, knee, or hip). The Fitts-like paradigm was used with two target amplitudes and six indices of difficulty (ID) for each of the four variables. This was done to explore the robustness of kinematic postural synergies. A speed accuracy trade-off was observed in all feedback conditions such that the movement time scaled with ID and the scaling differed between the two movement amplitudes. Principal-component (PC) analysis showed the existence of a single PC in the joint space that accounted for over 95% of the joint angle variance. Analysis within the uncontrolled manifold hypothesis has shown that data distributions in the joint angle space were compatible with stabilization of both trunk orientation and COM location. We conclude that trunk orientation and the COM location are stabilized by co-varied changes of the major joint angles during whole-body movements. Despite the strong effects of movement amplitude and ID on performance, the structure of the joint variance showed only minor dependence on these task parameters. The two kinematic synergies (co-varied changes in the joint angles that stabilized the COM location and trunk orientation) have proven to be robust over a variety of tasks. PMID- 16267117 TI - Macrophage migration inhibitory factor increases neuronal delayed rectifier K+ current. AB - Macrophage migration inhibitory factor (MIF) has widespread actions in the immune, endocrine, and nervous systems. Previously, we reported that increases in the intracellular levels of MIF depress the firing of hypothalamus/brain stem neurons in culture, including the chronotropic actions of angiotensin II. The objective of this study was to investigate the effects of MIF on delayed rectifier K+ current (I(Kv)), one of the component currents whose activity contributes to neuronal firing. Intracellular perfusion of MIF (80 nM) into Sprague-Dawley rat neuronal cultures caused a significant increase in I(Kv), as measured by patch-clamp recordings. This effect was apparent by 3 min, and was maximal after 20-30 min. I(Kv) current density (pA/pF) increased from 31.58 +/- 2.36 in controls to 41.88 +/- 3.76 in MIF-treated neurons (mean +/- SE; n = 9; P < 0.01). MIF that had been inactivated by boiling did not alter I(Kv), and MIF neutralizing antibodies abolished the action of recombinant MIF (rMIF). The stimulatory effect of MIF on I(Kv) current density was mimicked by intracellular application of either P1S-MIF (80 nM) or the peptide MIF-(50-65) (0.8-8 microM), both of which harbor the thiol-protein oxidoreductase (TPOR) activity of the MIF molecule. Conversely, neither C60S-MIF (80 nM) nor the MIF homologue D-dopachrome tautomerase (80 nM), both of which lack TPOR activity, altered I(Kv). Finally, the increase in I(Kv) produced by rMIF was abolished by the superoxide scavenger Tiron (1 mM). These studies indicate that the neuronal action of MIF includes a stimulatory action on I(Kv) that may be mediated by a TPOR/superoxide-scavenging mechanism. PMID- 16267119 TI - Sensory and motor effects of etomidate anesthesia. AB - The effects of anesthesia with etomidate on the cellular mechanisms of sensory processing and sensorimotor coordination have been studied in the active electric sense of the mormyrid fish Gnathonemus petersii. Like many anesthetics, etomidate is known to potentiate GABA(A) receptors, but little is known about the effects on sensory processing at the systems level. A better understanding is necessary for experimental studies of sensory processing, in particular regarding possible effects on the dynamic structure of excitatory and inhibitory receptive fields and to improve the knowledge of the mechanisms of anesthesia in general. Etomidate slowed the electromotor discharge rhythm, probably because of feedback inhibition at the premotor level, but did not alter the structure of the electromotor command. Sensory translation through primary afferents projecting to the cerebellum-like electrosensory lobe (ELL) was not changed. However, central interneurons and projection neurons were hyperpolarized under etomidate, and their spiking activity was reduced. Although the spatial extent and the center/surround organization of sensory receptive fields were not changed, initial excitatory responses were followed by prolonged inhibition. Corollary discharge input to ELL was maintained, and the temporal sequence of excitatory and inhibitory components of this descending signal remained intact. Later inhibitory corollary discharge responses were prolonged by several hundred milliseconds. The result was that excitatory reafferent sensory input was conserved with enhanced precision of timing, whereas background activity was greatly reduced. Anti-Hebbian synaptic plasticity evoked by association of sensory and corollary discharge input was still present under anesthesia, and differences compared with the nonanesthetized condition are discussed. PMID- 16267120 TI - Enhancement of spontaneous and heat-evoked activity in spinal nociceptive neurons by the endovanilloid/endocannabinoid N-arachidonoyldopamine (NADA). AB - N-arachidonoyldopamine (NADA) is an endogenous molecule found in the nervous system that is capable of acting as a vanilloid agonist via the TRPV1 receptor and as a cannabinoid agonist via the CB1 receptor. Using anesthetized rats, we investigated the neural correlates of behavioral thermal hyperalgesia produced by NADA. Extracellular single cell electrophysiology was conducted to assess the effects of peripheral administration of NADA (i.pl.) on nociceptive neurons in the dorsal horn of the spinal cord. Injection of NADA in the hindpaw caused increased spontaneous discharge of spinal nociceptive neurons compared with injection of vehicle. The neurons also displayed magnified responses to application of thermal stimuli ranging from 34 to 52 degrees C. NADA-induced neural hypersensitivity was dose dependent (EC50 = 1.55 microg) and TRPV1 dependent, as the effect was abolished by co-administration of the TRPV1 antagonist 5'-iodoresiniferatoxin (I-RTX). In contrast, co-administration of the CB1 antagonist SR 141716A did not attenuate this effect. These results suggest that the enhanced responses of spinal nociceptive neurons likely underlie the behavioral thermal hyperalgesia and implicate a possible pain-sensitizing role of endogenous NADA mediated by TRPV1 in the periphery. PMID- 16267121 TI - Role of anticipation in schizophrenia-related pursuit initiation deficits. AB - Schizophrenia patients exhibit several smooth pursuit abnormalities including poor pursuit initiation. Velocity discrimination is also impaired and is correlated with pursuit initiation performance-suggesting that pursuit deficits are related to impairments in processing velocity information. Studies suggest that pursuit initiation is influenced by prior target motion information and/or expectations and that this is likely caused by expectation-based changes in the perceptual inputs to the pursuit system. We examined whether poor pursuit initiation in schizophrenia results from inaccurate encoding of immediate velocity signals, or whether these deficits reflect a failure to use prior target motion information to "optimize" the response. Twenty-eight patients and 24 controls performed an adapted version of a "remembered pursuit task." Trials consisted of a series of target motions, the first of which occurred unexpectedly, followed by four to seven identical targets each preceded by an auditory cue and a "catch target" in which a cue was given followed by target extinction. Initiation eye velocity in response to unexpected, first targets was similar in the patient and control groups. In contrast, patients showed lower eye velocity in response to repeated, cued targets compared with controls. Patients also showed reduced eye velocity in response to catch targets. Reduction in pursuit latency across repeated targets was less robust in patients. Results suggest that processing of immediate velocity information is unaffected in schizophrenia and that pursuit initiation deficits reflect an inability to accurately generate, store, and/or access "remembered" velocity signals. PMID- 16267122 TI - Synapse elimination in the corticospinal projection during the early postnatal period. AB - In corticospinal synapses reconstructed in vitro by slice co-culture, we previously showed that the synapses were distributed across the gray matter at 6 7 days in vitro (DIV). Thereafter, they began to be eliminated from the ventral side, and dorsal-dominant distribution was nearly complete at 11-12 DIV. The synapse elimination is associated with retraction of the corticospinal (CS) terminals. We studied whether this specific type of synapse elimination is a physiological phenomenon rather than in vitro artifact. The rat corticospinal tract was stimulated at the medullary pyramid, and field potentials were recorded at the cervical cord along an 200-microm interval lattice on the axial plane. Clearly defined negative field potential were identified as field excitatory postsynaptic potentials (fEPSPs) generated by corticospinal synapses. They were recorded from the entire spinal gray matter at postnatal day 7 (P7). These negative fEPSPs reversed to positive in the most ventrolateral part at P8. Reversal extended to the more mediodorsal area at P10, indicative of progressive synapse elimination in the ventrolateral area. To verify that regression of the axons in vivo paralleled the changes in spatial distribution of fEPSPs as observed in vitro, corticospinal axons were anterogradely labeled. Redistribution of the labeled terminals closely paralleled the fEPSP distribution, being present in the ventrolateral spinal cord at P7, decreased at P8, further deceased at P10, but unchanged at P11. Furthermore, double immunostaining for labeled terminals and synaptophysin observed under a confocal microscope suggests that corticospinal fibers at P7 possess presynaptic structures in the ventrolateral area as well as the dorsomedial area. These findings suggest that corticospinal synapses are widely formed in the spinal gray matter at P7, are rapidly eliminated from the ventrolateral side from P8 to P10, a time-course very similar to that observed in vitro, and are associated with axonal regression. PMID- 16267124 TI - Rho-kinase as a molecular target for insulin resistance and hypertension. AB - Rho-kinase plays an important role in hypertension and is reported to interfere with insulin signaling through serine phosphorylation of insulin receptor substrate-1 (IRS-1) in cultured vascular smooth muscle cells. We therefore examined the role of Rho-kinase in the development of insulin resistance in Zucker obese rats. In skeletal muscles and aortic tissues of Zucker obese rats, activation of RhoA/Rho-kinase was observed. Long-term Rho-kinase inhibition by 4 wk treatment with fasudil (a Rho-kinase inhibitor) not only reduced blood pressure but corrected glucose and lipid metabolism, with improvement in serine phosphorylation of IRS-1 and insulin signaling in skeletal muscles. Direct visualization of skeletal muscle arterioles with an intravital CCD videomicroscope demonstrated that both acetylcholine- and sodium nitroprusside induced vasodilations were blunted, which were restored by the fasudil treatment. Furthermore, both fasudil and Y-27632 prevented the serine phosphorylation of IRS 1 induced by insulin and/or tumor necrosis factor-alpha in skeletal muscle cells. Collectively, Rho-kinase is responsible for the impairment of insulin signaling and may constitute a critical mediator linking between metabolic and hemodynamic abnormalities in insulin resistance. PMID- 16267123 TI - Early signaling responses to divergent exercise stimuli in skeletal muscle from well-trained humans. AB - Skeletal muscle from strength- and endurance-trained individuals represents diverse adaptive states. In this regard, AMPK-PGC-1alpha signaling mediates several adaptations to endurance training, while up-regulation of the Akt-TSC2 mTOR pathway may underlie increased protein synthesis after resistance exercise. We determined the effect of prior training history on signaling responses in seven strength-trained and six endurance-trained males who undertook 1 h cycling at 70% VO2peak or eight sets of five maximal repetitions of isokinetic leg extensions. Muscle biopsies were taken at rest, immediately and 3 h postexercise. AMPK phosphorylation increased after cycling in strength-trained (54%; P<0.05) but not endurance-trained subjects. Conversely, AMPK was elevated after resistance exercise in endurance- (114%; P<0.05), but not strength-trained subjects. Akt phosphorylation increased in endurance- (50%; P<0.05), but not strength-trained subjects after cycling but was unchanged in either group after resistance exercise. TSC2 phosphorylation was decreased (47%; P<0.05) in endurance-trained subjects following resistance exercise, but cycling had little effect on the phosphorylation state of this protein in either group. p70S6K phosphorylation increased in endurance- (118%; P<0.05), but not strength-trained subjects after resistance exercise, but was similar to rest in both groups after cycling. Similarly, phosphorylation of S6 protein, a substrate for p70 S6K, was increased immediately following resistance exercise in endurance- (129%; P<0.05), but not strength-trained subjects. In conclusion, a degree of "response plasticity" is conserved at opposite ends of the endurance-hypertrophic adaptation continuum. Moreover, prior training attenuates the exercise specific signaling responses involved in single mode adaptations to training. PMID- 16267125 TI - Purinoceptors are involved in the induction of an osmolyte permeability in malaria-infected and oxidized human erythrocytes. AB - In human erythrocytes, infection by the malaria parasite Plasmodium falciparum or oxidative stress induces a new organic osmolyte and anion permeability. To examine a role for autocrine purinoceptor signaling during this induction process, erythrocytic purinoceptor expression, and ATP release were determined. Furthermore, using pharmacological and genetic approaches the dependence on purinoceptor signaling of osmolyte permeability and Plasmodium development, both in vitro and in vivo, were assessed. Extracellular ATP did not induce an osmolyte permeability in non-infected or non-oxidized erythrocytes. ATP and other purinoceptor agonists increased the induction of osmolyte permeability during infection or oxidation as measured by isosmotic hemolysis and patch-clamp recording. Purinoceptor antagonists and apyrase decreased the induced permeability. The observed pharmacology suggested the involvement of P2Y purinoceptors. Accordingly, human erythrocytes expressed P2Y1 protein. Moreover, P2Y1-deficient mouse erythrocytes exhibited a delayed appearance of the osmolyte permeability during P. berghei infection- or oxidation compared with wild-type erythrocytes. Furthermore, the nonspecific purinoceptor antagonist suramin decreased in vitro growth and DNA/RNA amplification of P. falciparum in human erythrocytes and decreased in vivo growth of P. berghei. P. berghei developed slower in P2Y1-deficient mice in vivo compared with wild-type animals. In conclusion, induction of the osmolyte permeability in Plasmodium-infected erythrocytes involves autocrine purinoceptor signaling. PMID- 16267126 TI - Comparing cost/utility of giving an aromatase inhibitor as monotherapy for 5 years versus sequential administration following 2-3 or 5 years of tamoxifen as adjuvant treatment for postmenopausal breast cancer. AB - BACKGROUND: Several studies have shown aromatase inhibitors administered as monotherapy or sequentially to tamoxifen to improve relapse-free survival in postmenopausal women with early breast cancer. Any difference in cost/utility between the strategies may be of importance to therapy selection. METHODS: Cost/utility was compared between the different regimens based on the theoretical assumption that costs, benefits and side-effects were similar for each drug and independent of whether it was administered as monotherapy or sequentially. RESULTS: Tamoxifen for 2-3 years followed by an aromatase inhibitor for 3 or 2 years provided the lowest cost/quality-adjusted life years (QALY) estimates, while administration of an aromatase inhibitor subsequent to 5 years on tamoxifen provided the highest values. The difference between strategies increased with patient age. Cost/QALY estimates were sensitive to an increase in hip fracture risk and to cost reductions due to relapse prevention. Adding oral bisphosphonates increased costs moderately. CONCLUSIONS: While tamoxifen for 2-3 years followed by an aromatase inhibitor provided the lowest cost/QALY estimates, a further improvement of relapse-free survival of 1% if the aromatase inhibitor is given up front provides an acceptable cost/QALY. In contrast, additional benefits achieved by administering an aromatase inhibitor subsequent to 5 years of tamoxifen provided unacceptable costs. PMID- 16267127 TI - Assessment of open and incomplete bite correction by incisor overlap and optical density of polyvinyl siloxane bite registration. AB - Open bite (OB) is a generalized term, which could incorporate subgroups that react differently to vertical correction. The objectives of the present study were to detect vertical treatment changes in incomplete bite (IB: inter-incisor overlap with no lower incisor contact with teeth or palate) and OB (no inter incisor overlap) groups compared with a complete bite (CB: inter-incisor overlap with full lower incisor contact with teeth or palate) control group, to evaluate treatment response of the central and lateral incisors, and to study the vertico sagittal interaction. Dental casts were taken at three time points, pre treatment, post-treatment, and after one year of retention, from 54 Class II patients (22 males and 32 females with a mean age of 11 years 6 months) divided into three groups: CB (n = 21), IB (n = 18) and OB (n = 15). Measurements included incisor overlap (mm) and optical density (OD/mm2) of occlusal bite registration made of polyvinl siloxane. Both CB and IB groups demonstrated post retention bite opening. However, bite opening in the CB group was three times greater than that in the IB group (e.g. lower lateral = -1.42 mm, 118 OD/mm2 versus -0.40 mm, 107 OD/mm2). Conversely, the OB group showed a significant (P < 0.001) bite closure (e.g. lower lateral = 1.30 mm, -377 OD/mm2). Overjet changes affected OD measurements, causing diversity in OD and millimetric measurements of the lateral incisors in the IB group. In conclusion, the OB group demonstrated a significant stable vertical correction; a post-treatment non-contact inter incisor relationship was determined by a vertico-sagittal relapse; and full compensation of an IB was not possible. PMID- 16267128 TI - Evaluation of the functional diffusion map as an early biomarker of time-to progression and overall survival in high-grade glioma. AB - Diffuse malignant gliomas, the most common type of brain tumor, carry a dire prognosis and are poorly responsive to initial treatment. The response to treatment is typically evaluated by measurements obtained from radiographic images several months after the start of treatment; therefore, an early biomarker of tumor response would be useful for making early treatment decisions and for prognostic information. Thirty-four patients with malignant glioma were examined by diffusion MRI before treatment and 3 weeks later. These images were coregistered, and differences in tumor-water diffusion values were calculated as functional diffusion maps (fDM), which were correlated with the radiographic response, time-to-progression (TTP), and overall survival (OS). Changes in fDM at 3 weeks were closely associated with the radiographic response at 10 weeks. The percentage of the tumor undergoing a significant change in the diffusion of water (V(T)) was different between patients with progressive disease (PD) vs. stable disease (SD) (P < 0.001). Patients classified as PD by fDM analysis at 3 weeks were found to have a shorter TTP compared with SD (median TTP, 4.3 vs. 7.3 months; P < 0.04). By using fDM, early patient stratification also was correlated with shorter OS in the PD group compared with SD patients (median survival, 8.0 vs. 18.2 months; P < 0.01). On the basis of fDM, tumor assessment provided an early biomarker for response, TTP, and OS in patients with malignant glioma. Further evaluation of this technique is warranted to determine whether it may be useful in the individualization of treatment or evaluation of the response in clinical protocols. PMID- 16267129 TI - Computational docking and solution x-ray scattering predict a membrane interacting role for the histone domain of the Ras activator son of sevenless. AB - The Ras-specific nucleotide exchange factor son of sevenless (SOS) is a large, multidomain protein with complex regulation, including a Ras-dependent allosteric mechanism. The N-terminal segment of SOS, the histone domain, contains two histone folds, which is highly unusual for a cytoplasmic protein. Using a combination of computational docking, small-angle x-ray scattering, mutagenesis, and calorimetry, we show that the histone domain folds into the rest of SOS and docks onto a helical linker that connects the pleckstrin-homology (PH) and Dbl homology (DH) domains of SOS to the catalytic domain. In this model, a positively charged surface region on the histone domain is positioned so as to provide a fourth potential anchorage site on the membrane for SOS in addition to the PH domain, the allosteric Ras molecule, and the C-terminal adapter-binding site. The histone domain in SOS interacts with the helical linker, using a region of the surface that in nucleosomes is involved in histone tetramerization. Adjacent surface elements on the histone domain that correspond to the DNA-binding surface of nucleosomes form the predicted interaction site with the membrane. The orientation and position of the histone domain in the SOS model implicates it as a potential mediator of membrane-dependent activation signals. PMID- 16267130 TI - The antiinflammatory effect of laminar flow: the role of PPARgamma, epoxyeicosatrienoic acids, and soluble epoxide hydrolase. AB - We previously reported that laminar flow activates peroxisome proliferator activated receptor gamma (PPARgamma) in vascular endothelial cells in a ligand dependent manner that involves phospholipase A2 and cytochrome P450 epoxygenases. In this study, we investigated whether epoxyeicosatrienoic acids (EETs), the catalytic products of cytochrome P450 epoxygenases, are PPARgamma ligands. Competition and direct binding assays revealed that EETs bind to the ligand binding domain of PPARgamma with K(d) in the microM range. In the presence of adamantyl-ureido-dodecanoic acid (AUDA), a soluble epoxide hydrolase (sEH) specific inhibitor, EETs increased PPARgamma transcription activity in endothelial cells and 3T3-L1 preadipocytes. Inclusion of AUDA in the perfusing media enhanced, but overexpression of sEH reduced, the laminar flow-induced PPARgamma activity. Furthermore, laminar flow augmented cellular levels of EETs but decreased sEH at the levels of mRNA, protein, and activity. Blocking PPARgamma by GW9662 abolished the EET/AUDA-mediated antiinflammatory effect, which indicates that PPARgamma is an effector of EETs. PMID- 16267131 TI - Economic benefits of biodiversity exceed costs of conservation at an African rainforest reserve. AB - Economic research on biodiversity conservation has focused on the costs of conservation reserves and the benefits of intact ecosystems; however, no study has simultaneously considered the costs and benefits of species diversity, a fundamental component of biodiversity. We quantified the costs and benefits of avian biodiversity at a rainforest reserve in Uganda through a combination of economic surveys of tourists, spatial land-use analyses, and species-area relationships. Our results show that revising entrance fees and redistributing ecotourism revenues would protect 114 of 143 forest bird species (80%) under current market conditions. This total would increase to 131 species (approximately 90%) if entrance fees were optimized to capture the tourist's willingness to pay for forest visits and the chance of seeing increased numbers of bird species. In contrast, the cost of purchasing agricultural land for ecological rehabilitation of the avian habitat would be economically prohibitive. These results suggest that local biodiversity markets could play a positive role in tropical conservation strategies if the appropriate institutions for redistribution can be developed. PMID- 16267132 TI - Relation between the Widom line and the dynamic crossover in systems with a liquid-liquid phase transition. AB - We investigate, for two water models displaying a liquid-liquid critical point, the relation between changes in dynamic and thermodynamic anomalies arising from the presence of the liquid-liquid critical point. We find a correlation between the dynamic crossover and the locus of specific heat maxima C(P)(max) ("Widom line") emanating from the critical point. Our findings are consistent with a possible relation between the previously hypothesized liquid-liquid phase transition and the transition in the dynamics recently observed in neutron scattering experiments on confined water. More generally, we argue that this connection between C(P)(max) and dynamic crossover is not limited to the case of water, a hydrogen bond network-forming liquid, but is a more general feature of crossing the Widom line. Specifically, we also study the Jagla potential, a spherically symmetric two-scale potential known to possess a liquid-liquid critical point, in which the competition between two liquid structures is generated by repulsive and attractive ramp interactions. PMID- 16267133 TI - Protein folding by distributed computing and the denatured state ensemble. AB - The distributed computing (DC) paradigm in conjunction with the folding@home (FH) client server has been used to study the folding kinetics of small peptides and proteins, giving excellent agreement with experimentally measured folding rates, although pathways sampled in these simulations are not always consistent with the folding mechanism. In this study, we use a coarse-grain model of protein L, whose two-state kinetics have been characterized in detail by using long-time equilibrium simulations, to rigorously test a FH protocol using approximately 10,000 short-time, uncoupled folding simulations starting from an extended state of the protein. We show that the FH results give non-Poisson distributions and early folding events that are unphysical, whereas longer folding events experience a correct barrier to folding but are not representative of the equilibrium folding ensemble. Using short-time, uncoupled folding simulations started from an equilibrated denatured state ensemble (DSE), we also do not get agreement with the equilibrium two-state kinetics because of overrepresented folding events arising from higher energy subpopulations in the DSE. The DC approach using uncoupled short trajectories can make contact with traditionally measured experimental rates and folding mechanism when starting from an equilibrated DSE, when the simulation time is long enough to sample the lowest energy states of the unfolded basin and the simulated free-energy surface is correct. However, the DC paradigm, together with faster time-resolved and single molecule experiments, can also reveal the breakdown in the two-state approximation due to observation of folding events from higher energy subpopulations in the DSE. PMID- 16267134 TI - An improved reverse genetics system for influenza A virus generation and its implications for vaccine production. AB - The generation of vaccines for highly pathogenic avian influenza viruses, including those of the H5N1 subtype, relies on reverse genetics, which allows the production of influenza viruses from cloned cDNA. In the future, reverse genetics will likely be the method of choice for the generation of conventional influenza vaccine strains because gene reassortment by more traditional methods is cumbersome. Established systems for the artificial generation of influenza A viruses require transfection of cells with the eight to 12 plasmids that provide the eight influenza viral RNAs as well as the polymerase and nucleoproteins of the virus. However, cell lines appropriate for human vaccine production (e.g., Vero cells) cannot be transfected with high efficiencies. To overcome these problems, we established a reverse genetics system in which the eight RNA polymerase I transcription cassettes for viral RNA synthesis are combined on one plasmid. Similarly, two cassettes encoding the hemagglutinin and neuraminidase segments and six cassettes encoding the remaining proteins were combined. We also combined three RNA polymerase II transcription cassettes for the expression of the polymerase subunits. By combining these cassettes, we reduced the number of plasmids required for virus generation significantly and produced influenza A virus in Vero cells with higher efficiency than with the traditional 12 plasmid system. This new system is thus suitable for influenza virus vaccine production and may be applicable to other reverse genetics systems that rely on the introduction of several plasmids into eukaryotic cells. PMID- 16267135 TI - Biofilm formation and phenotypic variation enhance predation-driven persistence of Vibrio cholerae. AB - Persistence of the opportunistic bacterial pathogen Vibrio cholerae in aquatic environments is the principal cause for seasonal occurrence of cholera epidemics. This causality has been explained by postulating that V. cholerae forms biofilms in association with animate and inanimate surfaces. Alternatively, it has been proposed that bacterial pathogens are an integral part of the natural microbial food web and thus their survival is constrained by protozoan predation. Here, we report that both explanations are interrelated. Our data show that biofilms are the protective agent enabling V. cholerae to survive protozoan grazing while their planktonic counterparts are eliminated. Grazing on planktonic V. cholerae was found to select for the biofilm-enhancing rugose phase variant, which is adapted to the surface-associated niche by the production of exopolymers. Interestingly, grazing resistance in V. cholerae biofilms was not attained by exopolymer production alone but was accomplished by the secretion of an antiprotozoal factor that inhibits protozoan feeding activity. We identified that the cell density-dependent regulator hapR controls the production of this factor in biofilms. The inhibitory effect of V. cholerae biofilms was found to be widespread among toxigenic and nontoxigenic isolates. Our results provide a mechanistic explanation for the adaptive advantage of surface-associated growth in the environmental persistence of V. cholerae and suggest an important contribution of protozoan predation in the selective enrichment of biofilm forming strains in the out-of-host environment. PMID- 16267136 TI - An IL-7-dependent rebound in thymic T cell output contributes to the bone loss induced by estrogen deficiency. AB - The bone wasting induced by estrogen deficiency is, in part, a consequence of increased T cell production of the osteoclastogenic cytokine TNF-alpha. This phenomenon is due to an expansion of T cells, but the responsible mechanism is unknown. We now show that ovariectomy (ovx) disregulates T lymphopoiesis and induces bone loss by stimulating, through a rise in IL-7 levels, both thymic dependent differentiation of bone marrow-derived progenitors and thymic independent, peripheral expansion of mature T cells. Attesting to the relevance of the thymic effects, thymectomy decreases by approximately 50% the bone loss and the stimulation of T lymphopoiesis induced by ovx. In contrast, in vivo attenuation of the elevated IL-7 completely prevents the stimulation of T lymphopoiesis and the bone loss that follow ovx. Thus, the disruption of both T cell and bone homeostasis induced by ovx is mediated by IL-7 and due to both the thymic and extrathymic mechanisms. We conclude that IL-7 is a pivotal upstream target through which estrogen regulates hematopoietic and immune functions that are critical for bone homeostasis. PMID- 16267137 TI - The urotensin-II receptor antagonist palosuran improves pancreatic and renal function in diabetic rats. AB - Urotensin-II (U-II) is a cyclic peptide that acts through a specific G-protein coupled receptor, UT receptor. Urotensin-II and UT receptors have been described in pancreas and kidney, but their function is not well understood. We studied the effects of chronic treatment of diabetic rats with the orally active selective U II receptor antagonist palosuran. Streptozotocin treatment causes pancreatic beta cell destruction and leads to the development of hyperglycemia, dyslipidemia, and renal dysfunction. Long-term treatment of streptozotocin-induced diabetic rats with palosuran improved survival, increased insulin, and slowed the increase in glycemia, glycosylated hemoglobin, and serum lipids. Furthermore, palosuran increased renal blood flow and delayed the development of proteinuria and renal damage. The U-II system is unique in that it plays a role both in insulin secretion and in the renal complications of diabetes. Urotensin receptor antagonism might be a new therapeutic approach for the treatment of diabetes. PMID- 16267138 TI - Traditional Chinese medicines Wu Wei Zi (Schisandra chinensis Baill) and Gan Cao (Glycyrrhiza uralensis Fisch) activate pregnane X receptor and increase warfarin clearance in rats. AB - The traditional Chinese medicines (TCMs) are essential components of alternative medicines. Many TCMs are known to alter the expression of hepatic drug metabolizing enzymes and transporters. The molecular mechanism by which TCMs and/or their constituents regulate enzyme and transporter expression, however, has remained largely unknown. In this report, we show that two TCMs, Wu Wei Zi (Schisandra chinensis Baill) and Gan Cao (Glycyrrhiza uralensis Fisch), and their selected constituents activate the xenobiotic orphan nuclear receptor pregnane X receptor (PXR). Treatment with TCM extracts and the Schisandrol and Schisandrin constituents of Wu Wei Zi induced the expression of drug-metabolizing enzymes and transporters in reporter gene assays and in primary hepatocyte cultures. The affected enzymes and transporters include CYP3A and 2C isozymes and the multidrug resistance-associated protein 2. In transient transfection and reporter gene assays, the Schisandrin constituents of Wu Wei Zi had an estimated EC50 of 2 and 1.25 microM on hPXR and mPXR, respectively. Interestingly, mutations that were intended to alter the pore of the ligand-binding cavity of PXR had species specific effects on the activities of the individual Schisandrols and Schisandrins. In rats, the administration of Wu Wei Zi and Gan Cao increased the metabolism of the coadministered warfarin, reinforcing concerns involving the safe use of herbal medicines and other nutraceuticals to avoid PXR-mediated drug drug interactions. Meanwhile, the activation of PXR and induction of detoxifying enzymes provide a molecular mechanism for the hepatoprotective effects of certain TCMs. PMID- 16267139 TI - Hemispheric asymmetries in cortical thickness. AB - Using magnetic resonance imaging and computational cortical pattern matching methods, we analyzed hemispheric differences in regional gray matter thickness across the lateral and medial cortices in young, healthy adults (n = 60). In addition, we investigated the influence of gender on the degree of thickness asymmetry. Results revealed global and regionally specific differences between the two hemispheres, with generally thicker cortex in the left hemisphere. Regions with significant leftward asymmetry were identified in the precentral gyrus, middle frontal, anterior temporal and superior parietal lobes, while rightward asymmetry was prominent in the inferior posterior temporal lobe and inferior frontal lobe. On the medial surface, significant rightward asymmetries were observed in posterior regions, while significant leftward asymmetries were evident from the vicinity of the paracentral gyrus extending anteriorly. Asymmetry profiles were similar in both sexes, but hemispheric differences appeared slightly pronounced in males compared with females, albeit a few regions also indicated greater asymmetry in females compared with males. Hemispheric differences in the thickness of the cortex might be related to hemisphere specific functional specializations that are possibly related to behavioral asymmetries. PMID- 16267140 TI - Widespread recombination throughout Wolbachia genomes. AB - Evidence is growing that homologous recombination is a powerful source of genetic variability among closely related free-living bacteria. Here we investigate the extent of recombination among housekeeping genes of the endosymbiotic bacteria Wolbachia. Four housekeeping genes, gltA, dnaA, ftsZ, and groEL, were sequenced from a sample of 22 strains belonging to supergroups A and B. Sequence alignments were searched for recombination within and between genes using phylogenetic inference, analysis of genetic variation, and four recombination detection programs (MaxChi, Chimera, RDP, and Geneconv). Independent analyses indicate no or weak intragenic recombination in ftsZ, dnaA, and groEL. Intragenic recombination affects gltA, with a clear evidence of horizontal DNA transfers within and between divergent Wolbachia supergroups. Intergenic recombination was detected between all pairs of genes, suggesting either a horizontal exchange of a genome portion encompassing several genes or multiple recombination events involving smaller tracts along the genome. Overall, the observed pattern is compatible with pervasive recombination. Such results, combined with previous evidence of recombination in a surface protein, phage, and IS elements, support an unexpected chimeric origin of Wolbachia strains, with important implications for Wolbachia phylogeny and adaptation of these obligate intracellular bacteria in arthropods. PMID- 16267141 TI - Origin and evolution of the chloroplast trnK (matK) intron: a model for evolution of group II intron RNA structures. AB - The trnK intron of plants encodes the matK open reading frame (ORF), which has been used extensively as a phylogenetic marker for classification of plants. Here we examined the evolution of the trnK intron itself as a model for group II intron evolution in plants. Representative trnK intron sequences were compiled from species spanning algae to angiosperms, and four introns were newly sequenced. Phylogenetic analyses showed that the matK ORFs belong to the ML (mitochondrial-like) subclass of group II intron ORFs, indicating that they were derived from a mobile group II intron of the class. RNA structures of the introns were folded and analyzed, which revealed progressive RNA structural deviations and degenerations throughout plant evolution. The data support a model in which plant organellar group II introns were derived from bacterial-like introns that had "standard" RNA structures and were competent for self-splicing and mobility and that subsequently the ribozyme structures degenerated to ultimately become dependent upon host-splicing factors. We propose that the patterns of RNA structure evolution seen for the trnK intron will apply to the other group II introns in plants. PMID- 16267142 TI - The Trypanosoma cruzi L1Tc and NARTc non-LTR retrotransposons show relative site specificity for insertion. AB - The trypanosomatid protozoan Trypanosoma cruzi contains long autonomous (L1Tc) and short nonautonomous (NARTc) non-long terminal repeat retrotransposons. NARTc (0.25 kb) probably derived from L1Tc (4.9 kb) by 3'-deletion. It has been proposed that their apparent random distribution in the genome is related to the L1Tc-encoded apurinic/apyrimidinic endonuclease (APE) activity, which repairs modified residues. To address this question we used the T. cruzi (CL-Brener strain) genome data to analyze the distribution of all the L1Tc/NARTc elements present in contigs larger than 10 kb. This data set, which represents 0.91x sequence coverage of the haploid nuclear genome ( approximately 55 Mb), contains 419 elements, including 112 full-length L1Tc elements (14 of which are potentially functional) and 84 full-length NARTc. Approximately half of the full length elements are flanked by a target site duplication, most of them (87%) are 12 bp long. Statistical analyses of sequences flanking the full-length elements show the same highly conserved pattern upstream of both the L1Tc and NARTc retrotransposons. The two most conserved residues are a guanine and an adenine, which flank the site where first-strand cleavage is performed by the element encoded endonuclease activity. This analysis clearly indicates that the L1Tc and NARTc elements display relative site specificity for insertion, which suggests that the APE activity is not responsible for first-strand cleavage of the target site. PMID- 16267143 TI - An evolutionary space-time model with varying among-site dependencies. AB - It is now widely accepted that sites in a protein do not undergo independent evolutionary processes. The underlying assumption is that proteins are composed of conserved and variable linear domains, and thus rates at neighboring sites are correlated. In this paper, we comprehensively examine the performance of an autocorrelation model of evolutionary rates in protein sequences. We further develop a model in which the level of correlation between rates at adjacent sites is not equal at all sites of the protein. High correlation is expected, for example, in linear functional domains. On the other hand, when we consider nonlinear functional regions (e.g., active sites), low correlation is expected because the interaction between distant sites imposes independence of rates in the linear sequence. Our model is based on a hidden Markov model, which accounts for autocorrelation at certain regions of the protein and rate independence at others. We study the differences between the novel model and models which assume either independence or a fixed level of dependence throughout the protein. Using a diverse set of protein data sets we show that the novel model better fits most data sets. We further analyze the potassium-channel protein family and illustrate the relationship between the dependence of rates at adjacent sites and the tertiary structure of the protein. PMID- 16267144 TI - Overview of the National Health Educator Competencies Update Project, 1998-2004. AB - The National Health Educator Competencies Update Project (CUP), conducted during 1998-2004, addressed what health educators currently do in practice, the degree to which the role definition of the entry-level health educator is still up-to date, and the validation of advanced-level competencies. A 19-page questionnaire was sent to a representative sample of health educators in recognized practice settings in all states and the District of Columbia. A total of 4,030 health educators participated in the research (70.6% adjusted response rate) resulting in the largest national data set of its kind, with 1.6 million data points. The model derived from the research was hierarchical (7 areas of responsibility, 35 competencies, and 163 subcompetencies), with three levels of practice (Entry, Advanced 1, and Advanced 2) differentiated by degrees earned and years of experience. The findings affect professional preparation, credentialing, and professional development. PMID- 16267145 TI - Predictors of men's acceptance of modern contraceptive practice: study in rural Vietnam. AB - Studies have shown family planning adoption is likely to be more effective for women when men are actively involved. The transtheoretical model of behavior change was used to examine men's involvement in general contraception and intrauterine device (IUD) use by their wives. The study was carried out in rural Vietnam with 651 eligible participants. Cons of IUD use for men in precontemplation and contemplation/preparation were significantly higher than those in the action/maintenance stages, whereas the reverse was true for pros of IUD. The self-efficacy for convincing wife to have IUD in precontemplation was significantly lower than for those in higher stages. Women's education and ages, spontaneous recall of modern contraceptive method, cons for IUD, and self efficacy for contraception and for convincing wives to get IUD inserted (or continue use) were significant predictors of men's readiness to accept IUD. Interventions are targeted to reduce cons and increase self-efficacy for IUD use. PMID- 16267146 TI - Recreational gun use by California adolescents. AB - Most research on adolescents and firearms focuses on urban populations, handguns, and homicide. This study examines the prevalence and correlates of recreational gun use (RGU)--for hunting or target shooting--among 5,801 community-residing 12- to 17-year-old Californians. Data are from the first statewide California Health Interview Survey (CHIS), and person, design, and population weights were applied to the data. About one fifth (22.4%) of California adolescents report that they have gone hunting or target shooting. Nearly two thirds (62.8%) have hunted with a family member, typically (67.3%) their father. Recreational gun use among adolescents appears to be linked to a few basic demographic characteristics; most notably, male adolescents had an adjusted odds ratio of RGU nearly five times that of female adolescents. Some of the variables associated with RGU are consistent with those for violent gun use; differences, however, suggest that separate approaches to preventing firearm-related injury may be warranted. PMID- 16267147 TI - Acculturation, health protective sexual communication, and HIV/AIDS risk behavior among Hispanic women in a large midwestern city. AB - The relationships among acculturation, health protective sexual communication, and HIV/AIDS risk behaviors of Hispanic women are examined. Respondents represented 14 Latin American countries, a mean age of 29.5 years, and relatively low education and income levels. A significant correlation was found between higher non-Hispanic acculturation level and higher sexual communication with new partners. Higher non-Hispanic acculturation levels were also correlated with higher number of sexual partners in the prior 12 months. The higher the score for health protective sexual communication, the higher the reported frequency of condom use with new sexual partners in the past 12 months. Given the low number of sexual partners, HIV/AIDS risk for this group of women appears to be related to not knowing whether their partners had other sexual partners and to low frequency of condom use. Implications for health education are discussed. PMID- 16267148 TI - School tobacco control policies related to students' smoking and attitudes toward smoking: national survey results, 1999-2000. AB - The belief that schools can play a powerful role in preventing tobacco use among adolescents has led to the implementation of various tobacco-related policies and practices. This study examines the association between school policies regarding monitoring student behavior, severity of action taken for infraction of policies, and tobacco use by staff, and student smoking behavior and attitudes. Data on students' smoking behavior and attitudes were obtained from the 1999 and 2000 Monitoring the Future surveys of nationally representative samples of 8th-, 10th , and 12th-grade students. Data on school policies and practices were obtained from administrators in those same schools. Hierarchical analyses using HLM5 were conducted. Strictness of monitoring was significantly negatively associated with daily cigarette use by middle school students. Permitting staff to smoke was significantly positively associated with students' daily cigarette use and negatively with their disapproval of cigarette use. Policy implications are discussed. PMID- 16267149 TI - Australian adult consumers' beliefs about plant foods: a qualitative study. AB - This exploratory qualitative study examined consumers' perceived barriers and benefits of plant food (fruits, vegetables, grains, legumes, nuts, seeds) consumption and views on the promotion of these foods. Ten focus groups were conducted in Melbourne, Australia. Groups consisted of employees of various workplaces, community group members, university students, and inner-city residents. Health-related benefits predominated, particularly relating to the properties of plant foods (e.g., vitamins). Taste, variety, versatility, and environmental benefits were also considered important. The main barriers to eating plant foods were lack of knowledge and skills and length of preparation time. The poor quality of plant foods was also an issue for consumers. Awareness of the promotion of plant foods was generally high. Participants noted that promotions require a stronger practical emphasis with a focus on quick, easy-to prepare foods and meals. These findings provide insight into effective ways to promote a higher consumption of plant foods. PMID- 16267150 TI - The behavioral determinants of breast-feeding in The Netherlands: predictors for the initiation of breast-feeding. AB - The aim of this study was to evaluate the behavioral determinants of the initiation of breast-feeding at birth. The prospective cohort study used the attitude, social influence, self-efficacy (ASE) model in 373 pregnant women in five child health care centers. Prenatally, 72% of the women had the intention to breast-feed, and 73% actually started with breast-feeding at birth. Mothers who initiated breast-feeding differed in almost all the attitude, social influence, and self-efficacy determinants from others who initiated formula feeding. Intention was a very strong predictor of the initiation of breast-feeding. The components of the ASE model predicted the initiation of breast-feeding. Hence, the results may be used to tailor future interventions aimed at promoting breast feeding. PMID- 16267151 TI - The kidneys in the Bible: what happened? AB - The kidneys, always used in the plural (kelayot), are mentioned more than 30 times in the Bible. In the Pentateuch, the kidneys are cited 11 times in the detailed instructions given for the sacrificial offering of animals at the altar. Whereas those instructions were for purification ceremonies at the Temple, sacrificial offerings were made subsequently in seeking divine intervention for the relief of medical problems. In the books of the Bible that follow the Pentateuch, mostly in Jeremiah and Psalms, the human kidneys are cited figuratively as the site of temperament, emotions, prudence, vigor, and wisdom. In five instances, they are mentioned as the organs examined by God to judge an individual. They are cited either before or after but always in conjunction with the heart as mirrors of the psyche of the person examined. There is also reference to the kidneys as the site of divine punishment for misdemeanors, committed or perceived, particularly in the book of Job, whose suffering and ailments are legendary. In the first vernacular versions of the Bible in English, the translators elected to use the term "reins" instead of kidneys in differentiating the metaphoric uses of human kidneys from that of their mention as anatomic organs of sacrificial animals burned at the altar. This initial effort at linguistic purity or gentility has progressed further in recent versions of the Bible, in which the reins are now replaced by the soul or the mind. The erosion may have begun in the centuries that followed the writing of the Bible, when recognition of the kidneys as excretory organs deprived them of the ancient aura of mysterious organs hidden deep in the body but accessible to the look of God. At approximately the same time, Greek analytical philosophy argued that the brain, which is never mentioned in the Bible, was the most divine and sacred part of the body. This argument gained ground in the past century, when the functions of the brain were elucidated, and ultimately established in the 1960s, when salvaging the kidneys for transplantation necessitated a change in the definition of death as irreversible brain function. It is ironic that advances in understanding kidney function and in nephrology that made kidney transplantation feasible may have contributed, albeit indirectly, to the gradual elimination of the metaphoric mention of human kidneys in the Bible. PMID- 16267152 TI - Glomerular disease workshop. AB - Recent observations regarding intrinsic glomerular cell biology, particularly in the podocyte, have provided exciting new insights into potential pathogenic mechanisms of human glomerular disease. Although both immune and nonimmune mechanisms of glomerular injury have been studied previously, experimental models of disease and recent techniques that provide tools for molecular profiling show great promise for identifying glomerular disease biomarkers. Despite these recent advances, additional work in both basic and clinical studies of glomerular disease is needed to advance the field. Standardization of animal models of distinct forms of glomerular disease would likely facilitate the search for biomarkers. Several factors limit current efforts to implement clinical trials of glomerular disease. Identification of disease biomarkers, development of disease specific end points, and organization of collaborative clinical groups are critical for ultimately designing and implementing appropriately powered trials of glomerular disease. PMID- 16267153 TI - NIMA-related kinases defective in murine models of polycystic kidney diseases localize to primary cilia and centrosomes. AB - A key feature of the polycystic kidney diseases is aberrant cell proliferation, a consequence of dysfunctional ciliary signaling. The NIMA-related kinases (Nek) Nek1 and Nek8 carry the causal mutations of two of the eight established mouse models of polycystic kidneys. Nek proteins have roles in cell cycle and may contribute to coordinate regulation of cilia and cell-cycle progression. Herein is reported that in a mouse kidney epithelial cell line, mNek1 localizes to centrosomes in interphase and remains associated with the mitotic spindle pole during mitosis. In contrast, mNek8 localizes to the proximal region of the primary cilium and is not observed in dividing cells. Knockdown of mNek8 by siRNA does not affect ciliary assembly. Taken together with the phenotypes of the mutant mice, these data suggest that mNek1 and mNek8 provide links between cilia, centrosomes, and cell-cycle regulation. PMID- 16267154 TI - Hantavirus infection. PMID- 16267155 TI - Cystatin C and mortality risk in the elderly: the health, aging, and body composition study. AB - Kidney dysfunction is known to decrease life expectancy in the elderly. Cystatin C is a novel biomarker of kidney function that may have prognostic utility in older adults. The association of cystatin C with mortality was evaluated in a biracial cohort of black and white ambulatory elderly and compared with that of serum creatinine concentrations. The Health, Aging and Body Composition study is a cohort of well-functioning elderly that was designed to evaluate longitudinal changes in weight, body composition, and function. A total of 3075 participants who were aged 70 to 79 yr and had no disability were recruited at sites in Memphis, TN, and Pittsburgh, PA, between April 1997 and June 1998 with a follow up of 6 yr. At entry, the mean cystatin C was 1.05 mg/L and the mean creatinine was 1.06 mg/dl. After 6 yr of follow-up, 557 participants had died. The mortality rates in each ascending cystatin C quintile were 1.7, 2.7, 2.9, 3.1, and 5.4%/yr. After adjustment for demographic risk factors, comorbid health conditions, and inflammatory biomarkers (C-reactive protein, IL-6. and TNF-alpha), each quintile of cystatin C was significantly associated with increased mortality risk compared with the lowest: Hazard ratios (HR; 95% confidence intervals) quintile 1, -1.0 (referent); quintile 2, -1.74 (1.21 to 2.50); quintile 3, -1.51 (1.05 to 2.18); quintile 4, -1.49 (1.04 to 2.13); and quintile 5, -2.18 (1.53 to 3.10). These associations did not differ by gender or race. Results were consistent for cardiovascular and other-cause mortality, but not cancer mortality. Creatinine quintiles were not associated with mortality after multivariate adjustment (HR: 1.0 [referent], 1.00 [0.72 to 1.39], 0.95 [0.68 to 1.32], 1.11 [0.79 to 1.57], 1.16 [0.86 to 1.58]). Cystatin C is a strong, independent risk factor for mortality in the elderly. Future studies should investigate whether cystatin C has a role in clinical medicine. PMID- 16267157 TI - Chemokine receptor Ccr2 deficiency reduces renal disease and prolongs survival in MRL/lpr lupus-prone mice. AB - MRL/MpJ-Fas(lpr)/J (MRL/lpr) mice represent a well-established mouse model of human systemic lupus erythematosus. MRL/lpr mice homozygous for the spontaneous lymphoproliferation mutation (lpr) are characterized by systemic autoimmunity, massive lymphadenopathy associated with proliferation of aberrant T cells, splenomegaly, hypergammaglobulinemia, arthritis, and fatal immune complex mediated glomerulonephritis. It was reported previously that steady-state mRNA levels for the chemokine (C-C motif) receptor 2 (Ccr2) continuously increase in kidneys of MRL/lpr mice. For examining the role of Ccr2 for development and progression of immune complex-mediated glomerulonephritis, Ccr2-deficient mice were generated and backcrossed onto the MRL/lpr genetic background. Ccr2 deficient MRL/lpr mice developed less lymphadenopathy, had less proteinuria, had reduced lesion scores, and had less infiltration by T cells and macrophages in the glomerular and tubulointerstitial compartment. Ccr2-deficient MRL/lpr mice survived significantly longer than MRL/lpr wild-type mice despite similar levels of circulating immunoglobulins and comparable immune complex depositions in the glomeruli of both groups. Anti-dsDNA antibody levels, however, were reduced in the absence of Ccr2. The frequency of CD8+ T cells in peripheral blood was significantly lower in Ccr2-deficient MRL/lpr mice. Thus Ccr2 deficiency influenced not only monocyte/macrophage and T cell infiltration in the kidney but also the systemic T cell response in MRL/lpr mice. These data suggest an important role for Ccr2 both in the general development of autoimmunity and in the renal involvement of the lupus-like disease. These results identify Ccr2 as an additional possible target for the treatment of lupus nephritis. PMID- 16267156 TI - Nephrogenic factors promote differentiation of mouse embryonic stem cells into renal epithelia. AB - Embryonic stem (ES) cells have been induced to differentiate in vitro into a broad spectrum of specialized cell types, including hematopoietic, pancreatic, and neuronal cell types. Such ES-derived cells can provide a valuable source of progenitor cell types. Whereas undifferentiated ES cells can become integrated into a developing kidney and contribute to tubular epithelia, the ability to generate renal precursor cells in vitro has not been reported. This study used a combination of nephrogenic growth factors to differentiate ES cells into renal epithelial cells that are capable of integrating into a developing kidney with very high efficiency. Using a combination of retinoic acid, Activin-A, and Bmp7, cultured ES cells can be induced to express markers specific for the intermediate mesoderm, from which the kidneys arise. Embryoid bodies that are cultured in the presence of nephrogenic factors can respond to inductive signals and form epithelial structures in vitro. When injected into developing kidney rudiments, treated ES cells contribute to tubular epithelia with near 100% efficiency. These methods may facilitate the large-scale culture of renal epithelial precursor cells for a variety of applications. PMID- 16267158 TI - Hyperaldosteronemia and activation of the epithelial sodium channel are not required for sodium retention in puromycin-induced nephrosis. AB - Edema and ascites in nephrotic syndrome mainly result from increased Na+ reabsorption along connecting tubules and cortical collecting ducts (CCD). In puromycin aminonucleoside (PAN)-induced nephrosis, increased Na+ reabsorption is associated with increased activity of the epithelial sodium channel (ENaC) and Na+,K+-ATPase, two targets of aldosterone. Because plasma aldosterone increases in PAN-nephrotic rats, the aldosterone dependence of ENaC activation in PAN nephrosis was investigated. For this purpose, (1) the mechanism of ENaC activation was compared in nephrotic and sodium-depleted rats, and (2) ENaC activity in PAN-nephrotic rats was evaluated in the absence of hyperaldosteronemia. The mechanism of ENaC activation was similar in CCD from nephrotic and sodium-depleted rats, as demonstrated by (1) increased number of active ENaC evaluated by patch clamp, (2) recruitment of ENaC to the apical membrane determined by immunohistochemistry, (3) shift in the electrophoretic profile of gamma-ENaC, and (4) increased abundance of beta-ENaC mRNA. Corticosteroid clamp fully prevented all PAN-induced changes in ENaC but did not alter the development of a full-blown nephrotic syndrome with massive albuminuria, amiloride-sensitive sodium retention, induction of CCD Na+,K+ ATPase, and ascites. It is concluded that in PAN-nephrosis, (1) ENaC activation in CCD is secondary to hyperaldosteronemia, (2) sodium retention and induction of Na+,K+-ATPase in CCD are independent of hyperaldosteronemia, and (3) ENaC is not necessarily limiting for sodium reabsorption in the distal nephron. PMID- 16267159 TI - Activation of Galpha q-coupled signaling pathways in glomerular podocytes promotes renal injury. AB - The glomerular podocyte plays a key role in maintaining the integrity of the glomerular filtration barrier. This function may be regulated by activation of cell surface G protein-coupled receptors (GPCR). Studies suggest that podocytes express GPCR that are implicated in the pathogenesis of glomerular diseases. Common to these GPCR systems is activation of phospholipase C through the Gq alpha-subunit (Galpha q). For investigating the role of Galpha q-coupled signaling pathways in promoting renal injury in podocytes, a constitutively active Galpha q subunit (Galpha qQ > L) was expressed in glomerular podocytes using the mouse nephrin promoter. Transgenic (TG) mice demonstrated albuminuria as well as a decrease in both kidney mass and nephron number. By light microscopy, a portion of the TG mice had glomerular abnormalities, including focal to diffuse hypercellularity and segmental sclerosis. Consistent with injury promoting effects of Galpha qQ > L, there was a significant reduction in podocalyxin mRNA as well as nephrin mRNA and protein levels in glomeruli from TG mice compared with non-TG controls. Expression of the transgene also seemed to increase susceptibility to glomerular injury, because treatment with puromycin aminonucleoside enhanced proteinuria in TG mice compared with non-TG littermate controls (4.2 +/- 1.0 [TG] versus 1.6 +/- 0.3 [non-TG] mg/24 h; P = 0.0161). Thus, activation of Galpha q in glomerular podocytes caused alterations in glomerular histomorphology, albuminuria, decreased nephron mass, and reduced glomerular expression of both nephrin and podocalyxin as well as enhanced susceptibility to glomerular damage induced by puromycin aminonucleoside. It is speculated that Galpha q-coupled signaling cascades may be important effector pathways mediating renal injury. PMID- 16267160 TI - Projecting the number of patients with end-stage renal disease in the United States to the year 2015. AB - The size of the prevalent ESRD population in the United States increased dramatically during the 1990s, from 196,000 in 1991 to 382,000 in 2000. Incidence also increased considerably during the same period, from 53,000 to 93,000 per year. If previous trends in ESRD incidence and prevalence continue, then current levels of health care resources that are devoted to the care of these patients will eventually be unable to meet the demand. This study discusses a Markov model developed to predict ESRD incidence, prevalence, and mortality to the year 2015 and incorporating expected changes in age/race distributions, diabetes prevalence, ESRD incidence, and probability of death. The model predicted that by 2015 there will be 136,166 incident ESRD patients per year (lower/upper limits 110,989 to 164,550), 712,290 prevalent patients (595,046 to 842,761), and 107,760 ESRD deaths annually (96,068 to 118,220). Incidence and prevalence counts are expected to increase by 44 and 85%, respectively, from 2000 to 2015 and incidence and prevalence rates per million population by 32 and 70%, respectively. The financial and human resources that will be needed to care for these patients in 2015 will be considerably greater than in 2005. PMID- 16267161 TI - Computation in the olfactory system. AB - Computational models are increasingly essential to systems neuroscience. Models serve as proofs of concept, tests of sufficiency, and as quantitative embodiments of working hypotheses and are important tools for understanding and interpreting complex data sets. In the olfactory system, models have played a particularly prominent role in framing contemporary theories and presenting novel hypotheses, a role that will only grow as the complexity and intricacy of experimental data continue to increase. This review will attempt to provide a comprehensive, functional overview of computational ideas in olfaction and outline a computational framework for olfactory processing based on the insights provided by these diverse models and their supporting data. PMID- 16267162 TI - Differential covariation in taste responsiveness to bitter stimuli in rats. AB - Variation exists in the sensitivity of individual rodents and humans to different bitter tastants. An absence of uniform correlation in responsiveness to different bitter substances across individuals within a species suggests heterogeneity in the mechanisms underlying stimulus processing within this taste modality. Here, we examined taste responsiveness of individual rats to three bitter compounds (quinine hydrochloride, denatonium benzoate, and cycloheximide) in short-term lick tests to determine the magnitude of covariation among responses to these stimuli and infer commonalities in their receptor and neural mechanisms. Rats were tested with a given pair of bitter stimuli during three sessions comprising randomized trial blocks of six concentrations of each stimulus + deionized water. Psychophysical functions were generated for individual rats for respective stimulus pairs, and concentrations of each stimulus that produced equivalent lick suppression relative to water were correlated across animals. Behavioral taste responsiveness to quinine hydrochloride strongly covaried with responsiveness to denatonium benzoate (r = +0.82). Lick responsiveness to quinine was less robustly correlated with that to cycloheximide (r = +0.44), and denatonium and cycloheximide responses failed to correlate. These results imply substantial overlap in the bitter taste coding mechanisms for quinine and denatonium but some degree of independence in the mechanisms responsible for gustatory processing of cycloheximide. More generally, these data reinforce the notion that bitter taste processing is not a homogeneous event. PMID- 16267163 TI - Electroantennographic resolution of pulsed pheromone plumes in two species of moths with bipectinate antennae. AB - Trains of 20-ms-duration pulses of pheromone were delivered at rates of 1-33 Hz to antennal preparations of males of Bombyx mori and Lymantria dispar, two moth species with bipectinate antennae. Resolution of rapidly pulsed plumes of pheromone was not compromised by a complex antennal morphology or by moderate changes in wind speed (25-50 cm/s). Fourier analysis of the electroantennograms resolved the temporal structure of the signal at frequencies up to 25 Hz for B. mori and up to 5 Hz for L. dispar. The ability of these sensory structures to identify the original (unchanged) frequency of the pulse train is particularly noteworthy because air is slowed by about an order of magnitude as it passes through bipectinate antennae. Although an unchanging frequency in slowed airflow may be counterintuitive, this flow pattern, and its effects on odorant patch shape and spacing, is explained from fluid mechanical principles (i.e., the principle of continuity). An unchanging frequency suggests that as decelerating air passes through a bipectinate antenna, the slowed patches of odorant are stretched, thinned, and brought closer together by the same factor with which they are slowed. PMID- 16267164 TI - Exclusion of candidate genes for canine SRY-negative XX sex reversal. AB - In mammals, the Y-linked SRY gene is normally responsible for testis induction, yet testis development can occur in the absence of Y-linked genes, including SRY. The canine model of SRY-negative XX sex reversal could lead to the discovery of novel genes in the mammalian sex determination pathway. The autosomal genes causing testis induction in this disorder in dogs, humans, pigs, and horses are presently unknown. In goats, a large deletion is responsible for sex reversal linked to the polled (hornless) phenotype. However, this region has been excluded as being causative of the canine disorder, as have WT1 and DMRT1 in more recent studies. The purpose of this study was to determine whether microsatellite marker alleles near or within five candidate genes (GATA4, FOG2, LHX1, SF1, SOX9) are associated with the affected phenotype in a pedigree of canine SRY-negative XX sex reversal. Primer sequences flanking nucleotide repeats were designed within genomic sequences of canine candidate gene homologues. Fluorescence-labeled polymorphic markers were used to screen a subset of the multigenerational pedigree, and marker alleles were determined by software. Our results indicate that the mutation causing canine SRY-negative XX sex reversal in this pedigree is unlikely to be located in regions containing these candidates. PMID- 16267165 TI - Complex segregation analysis of canine hip dysplasia in German shepherd dogs. AB - Complex segregation analyses were carried out to clarify the mode of inheritance of canine hip dysplasia (CHD) in German shepherd dogs. Data were used from 8,567 animals examined for CHD from 20 families with three to four generations. The existence of a major gene in addition to polygenic gene effects was detected. In the present study, a mixed model with a dominant major gene effect seemed to be most probable for dichotomous encoding (0: dogs without signs of CHD; 1: dogs with borderline/slight to severe CHD). In addition, mixed major gene inheritance was shown for a binary trait where borderline was assigned to dogs scored free from CHD and for a trichotomously encoded trait (0: dogs without signs of CHD; 1: borderline CHD; 2: mild to severe CHD). Although only small frequencies were found for the unfavorable homozygotic genotype AA, the probability of the AB genotype was high in affected animals. Selection schemes to reduce the frequency of the allele A should therefore efficiently improve existing breeding programmes in German shepherd dogs. PMID- 16267167 TI - Canine DNA subjected to whole genome amplification is suitable for a wide range of molecular applications. AB - Molecular and genetic studies of canine disease phenotypes can be limited by the amount of DNA available for analysis. New methods have been developed to amplify the genomic DNA of a species producing large quantities of DNA from small starting amounts. Whole genome amplification (WGA) of DNA is now being used in human studies, although this technique has not been applied extensively in veterinary research. We evaluated WGA of canine DNA for suitability in a range of molecular tests. DNA from 93 canine blood extracted and 18 buccal swab samples was subjected to WGA using the GenomiPhi kit (Amersham). Genomic DNA was compared with WGA product using a range of techniques, including reference strand-mediated conformation analysis, denaturing high-performance liquid chromatography analysis, microsatellite genotyping, direct DNA sequencing, and single nucleotide polymorphism allelic discrimination. All samples amplified well, giving an average yield of 3 mug of DNA from 2.5 ng of starting material. Extremely high levels of experimental reproducibility and concordance were observed between source and WGA DNA samples for all analyses used: greater than 95% for blood extracted DNA and greater than 80% for buccal swab DNA. These studies clearly demonstrate the usefulness of WGA of canine DNA as a means of increasing DNA quantities for canine studies. This technique will have major implications for future veterinary research. PMID- 16267166 TI - Phylogenetic analysis of snow sheep (Ovis nivicola) and closely related taxa. AB - Based on mitochondrial cytochrome b gene sequence analysis, the history of true sheep (Ovis) began approximately 3.12 million years ago (MYA). The evolution of Ovis resulted in three generally accepted genetic groups: Argaliforms, Moufloniforms, and Pachyceriforms. The Pachyceriforms of the subgenus Pachyceros comprise the thin-horn sheep Ovis nivicola (snow sheep), Ovis dalli (Dall and Stone sheep), and Ovis canadensis (Rocky Mountain and desert bighorn). North America wild sheep (O. canadensis and O. dalli) evolved separately from Eurasian wild sheep and diverged from each other about 1.41 MYA. Ancestral stock that gave rise to snow sheep, Moufloniforms, and Argaliforms occurred 2.3 MYA, which then gave rise to two different extant lines of snow sheep that diverged from each other about 1.96 MYA. The more recent nivicola line is genetically closer to the North American wild sheep and may represent a close association during the refugium when Alaska and Siberia were connected by the Bering land bridge. The earlier period of evolution of the Pachyceriforms suggests they may have first evolved in Eurasia, the oldest ancestor then giving rise to North American wild sheep, and that a canadensis-like ancestor most likely gave rise to nivicola. Cytogenetic analysis further validates that the standard diploid number for modern nivicola is 52. PMID- 16267168 TI - Fecal DNA analysis for identifying species and sex of sympatric carnivores: a noninvasive method for conservation on the Tsushima Islands, Japan. AB - Fecal analysis is a useful tool for the investigation of food habits and species identity in mammals. However, it is generally difficult to identify the species based on the morphological features and contents of feces deposited by mammals of similar body size. Therefore we developed noninvasive DNA analysis methods using fecal samples for identification of the species and sex of four small sympatric carnivores living on the Tsushima Islands of Japan: the leopard cat (Felis bengalensis), Japanese marten (Martes melampus), Siberian weasel (Mustela sibirica), and feral cat (Felis catus). Based on DNA sequence data from previous phylogenetic studies, we designed species-specific primers for polymerase chain reaction (PCR) amplification of the partial mitochondrial cytochrome b gene (112 347 bp) to identify the species and primers for the partial SRY gene (135 bp) to determine the sex. Due to the adjustment of PCR conditions, those specific DNA fragments were successfully amplified and then applied for species and sex identification. Nucleotide sequences obtained from the PCR products corresponded with cytochrome b sequences of the carnivore species expected. The protocol developed could be a valuable tool in the management and conservation of the four carnivore species occurring on the Tsushima Islands. PMID- 16267170 TI - High diversity of the chicken growth hormone gene and effects on growth and carcass traits. AB - The chicken growth hormone (cGH) gene plays a crucial role in controlling growth and metabolism, leading to potential correlations between cGH polymorphisms and economic traits. In this study, DNA from four divergent chicken breeds were screened for single nucleotide polymorphisms (SNPs) in the cGH gene using denaturing high-performance liquid chromatography and sequencing. A total of 46 SNPs were identified, of which 4 were in the 5' untranslated region, 1 in the 3' untranslated region, 5 in exons (two of which are nonsynonymous), with the remaining 36 in introns. The nucleotide diversity in the cGH gene ( theta = 2.7 x 10(-3)) was higher than that reported for other chicken genes, even within the same breeds. The associations of five of these SNPs and their haplotypes with chicken growth and carcass traits were determined using polymerase chain reaction restriction fragment length polymorphism analysis in a F2 resource population cross of two of the four chicken breeds (White Recessive Rock and Xinghua). This analysis shows that, among other correlations, G+1705A was significantly associated with body weight at all ages measured, shank length at three of four ages measured, and average daily gain within weeks 0 to 4. Thus, this cGH polymorphism, or another polymorphism that is in linkage disequilibrium with G+1705A, appears to correspond to a significant growth-related quantitative trait locus difference between the two breeds used to construct the resource population. PMID- 16267169 TI - Genetic diversity within and among four South European native horse breeds based on microsatellite DNA analysis: implications for conservation. AB - In the present study, genetic analyses of diversity and differentiation were performed on four Basque-Navarrese semiferal native horse breeds. In total, 417 animals were genotyped for 12 microsatellite markers. Mean heterozygosity was higher than in other horse breeds, surely as a consequence of management. Although the population size of some of these breeds has declined appreciably in the past century, no genetic bottleneck was detected in any of the breeds, possibly because it was not narrow enough to be detectable. In the phylogenetic tree, the Jaca Navarra breed was very similar to the Pottoka, but appeared to stand in an intermediate position between this and the meat breeds. Assuming that Pottoka is the breed less affected by admixture, the others gradually distanced themselves from it through varying influences from outside breeds, among other factors. In a comparative study with other breeds, the French breeds Ardanais, Comtois, and Breton were the closest to the four native breeds. Three different approaches for evaluating the distribution of genetic diversity were applied. The high intrabreed variability of Euskal Herriko Mendiko Zaldia (EHMZ) was pointed out in these analyses. In our opinion, cultural, economic, and scientific factors should also be considered in the management of these horse breeds. PMID- 16267171 TI - The many faces of the copper metabolism protein MURR1/COMMD1. AB - Copper is an essential transition metal but is toxic in excess; therefore, its metabolism needs to be tightly regulated. Defects in the regulation of copper can lead to various disorders characterized by copper deficiency or copper excess. Recently, we characterized the COMMD1 (previously MURR1) gene as the defective gene in canine copper toxicosis. The molecular functions of COMMD1 remain unknown, but significant progress has been made in identifying the cellular processes in which COMMD1 participates, through the identification of proteins interacting with COMMD1. This review discusses how COMMD1 functions as a regulator of not only copper homeostasis but also sodium transport and the NF kappaB signaling pathway. We outline the possible mechanisms through which COMMD1 exerts these newly identified functions. PMID- 16267173 TI - Vitamin D and fracture prevention--treatment still indicated but clarification needed. PMID- 16267174 TI - Reversible dementia--the implications of a fall in prevalence. PMID- 16267175 TI - Chronic kidney disease and older people--implications of the publication of the Part 2 of the National Service Framework for Renal Services. PMID- 16267176 TI - Mental capacity assessments and discharge decisions. PMID- 16267177 TI - The future of geriatric medicine in an era of patient choice. PMID- 16267178 TI - Accelerometers in rehabilitation medicine for older adults. AB - Recent technological developments have led to the production of inexpensive, miniature accelerometer sensors with potential for use in a clinical setting. These sensors can provide reliable information on mobility and objective measurement of gait. They are currently used mainly in a research setting; however, with recent advances, incorporation into clinical practice is possible. For illustrative purposes this paper describes some current applications of accelerometers in gait and balance evaluation, falls risk assessment and mobility monitoring. Accelerometers provide quantitative measures of gait, they are capable of identifying specific gait changes in older adults and in fallers and can be used to objectively quantify ambulatory activity levels. Accelerometers have many potential uses in monitoring of patients in rehabilitation. They provide an added objective and quantitative dimension to gait analysis when combined with clinical assessment. They have the potential in the future to stratify falls risk facilitating early initiation of appropriate therapeutic intervention, thus reducing further falls. The challenge facing clinicians and biomedical engineers is to further harness this technology making it part of everyday clinical practice. PMID- 16267179 TI - A systematic review of prevalence and incidence studies of dementia with Lewy bodies. AB - BACKGROUND: Substantial variation in the prevalence of Dementia with Lewy Bodies (DLB) has been reported with estimates ranging from 0 to 26.3% of all dementia cases, potentially making it the second most common dementia subtype. OBJECTIVES: The aim of this study was to review systematically and critically for the first time previous studies of the clinical prevalence and incidence of DLB in the population. METHODS: A systematic literature search was performed using PubMed. Selected articles had to describe an original study that provided a prevalence and/or incidence number for the whole population for DLB as defined by pre-set clinical criteria and findings. RESULTS: Six studies reporting the prevalence of DLB and one study reporting the incidence of DLB met the inclusion criteria. Prevalence estimates, depending on case criteria, range from 0 to 5% with regard to the general population, and from 0 to 30.5% of all dementia cases. The only estimate for DLB incidence is 0.1% a year for the general population and 3.2% a year for all new dementia cases. The number of available studies was too small to hypothesise on the potential effect of age, sex and genetic background on the results. CONCLUSIONS: Although the literature on the prevalence and incidence of DLB is limited, there is a general consensus that DLB must be considered in the range of neurodegenerative conditions in the elderly. The move towards use of consensus criteria facilitates comparison and is welcome. Their application in a more routine way towards rigorously defined and selected study populations will lead to more comparable and generalisable studies in the future. PMID- 16267180 TI - Streamlining assessment and intervention in a falls clinic using the Timed Up and Go Test and Physiological Profile Assessments. AB - BACKGROUND: The Timed Up and Go Test (TUGT) has been recommended as a simple screening tool to identify those at risk of falling. However, subsequent detailed assessment is required to identify underlying falls risk factors to provide direction for optimal targeted intervention strategies. METHODS: 110 consecutive falls clinic patients underwent the TUGT, the Abbreviated Mental Test and the Physiological Profile Assessment (PPA), a validated tool for quantifying risk of falling based on a combination of physiological measures-contrast sensitivity, knee extension strength, proprioception, reaction time and postural sway. Regression analysis was used to determine how well the TUGT and presence of cognitive impairment could identify patients at high risk of falls as defined by the PPA. RESULTS: TUGT and cognitive status were found to be independent and significant predictors of PPA scores. These variables accounted for 21% of the variance in PPA scores (multiple R=0.47, P<0.001). The standardised beta weights were 0.403 for TUGT and 0.236 for cognitive status. A receiver-operator curve (ROC) indicated that 15 seconds in the TUGT was the optimal cut-point for identifying those with a high risk of falling: 70% of the total sample. CONCLUSIONS: The TUGT and a simple test of cognition can be used to streamline referrals in a high-risk population, allowing for more efficient use of available resources in clinical practice. A subsequent PPA provides quantification of risk and direction for tailored intervention. PMID- 16267181 TI - Endoscopic retrograde cholangiopancreatography in the elderly: a prospective and comparative study. AB - OBJECTIVE: In this study we aimed to compare the utility and safety of endoscopic retrograde cholangiopancreatography (ERCP) in elderly and younger groups of people. METHODS: The study involved 299 patients who underwent ERCP for the first time between March 2002 and January 2003. Group A consisted of 202 patients who were 69 years of age or younger and group B involved 97 patients who were 70 years of age or older. The patients were prospectively identified and data were gathered on them prospectively. Clinical and biochemical features, ERCP procedures, ERCP diagnosis, complications and length of hospital stay were compared between the two groups. RESULTS: Group B patients presented with more specific symptoms compared with group A. Laboratory findings were similar in both groups other than mean bilirubin levels, which were higher in the elderly group. Selective biliary cannulation was technically successful in 99% of both groups. Pre-cut papillotomy was performed in 49.5% of group A patients and 56.7% of group B patients. Choledocholithiasis was the most frequent diagnosis in both groups. Benign biliary stenosis was more frequent in group A and malignant biliary stenosis in group B. Among the malignant aetiologies, pancreatic carcinoma was the most common cancer in both groups. Post-procedural complications developed in 27 (9%) patients. There were 16 (7.9%) complications (six bleeding, five pancreatitis, three perforation and two cholangitis) in the 202 patients aged under 70 years. In comparison, 11 (11.3%) of the 97 patients aged over 70 years developed a complication (bleeding in six, pancreatitis in one, perforation in one, cholangitis in two and ileus in one). Twelve (nine group A and three group B) patients died within 5-30 days of the procedure. Length of hospital stay was comparable for both groups (15.53+/-12.43 days in group A and 14.84+/-11.56 days in group B). CONCLUSIONS: Diagnostic and therapeutic ERCP has similar outcomes in both elderly and younger patients. ERCP is effective and safe in elderly patients with naive papilla. The more invasive procedures like pre-cut papillotomy may be performed safely in the elderly. PMID- 16267183 TI - Renal transplantation in the elderly: does patient age determine the results? AB - BACKGROUND: Transplantation is the best treatment for patients with chronic renal failure, including the elderly. However, the patient's age was traditionally considered as a relative contraindication for it. OBJECTIVE: To compare the results of renal transplantation in patients over and under 60 years of age. METHODS: Analysis of 621 transplant recipients in Galicia (Spain) between 1996 and 2000, divided into two groups, according to age over 60 years (484) or under 60 years (137). The actuarial method, Kaplan-Meier curves, log-rank test and Cox proportional hazard model were used to study survival. RESULTS: Graft survival for those aged under 60 years was 82% and 70% at 1 and 5 years, while it was 73% and 56% for those over 60 years. However, censuring the deceased patients with a functioning graft, it was 84% and 76% for those aged under 60 years and 83% and 77% for those over 60 years. A total of 47% of the graft losses in the group over 60 years were due to the patient's death. Overall graft survival for all the patients was greater (P<0.0001) when the donor was under 60 years of age. CONCLUSIONS: Recipient age alone cannot be a criterion to exclude patients over 60 years from transplantation, since their lower survival is influenced by comorbidity and the donor's age. PMID- 16267182 TI - A whole system study of intermediate care services for older people. AB - BACKGROUND: Intermediate care (IC) services have been widely introduced in England and have the strategic objectives of reducing hospital and long-term care use. There is uncertainty about the clinical outcomes of these services and whether their strategic aims will be realised. SETTING: A metropolitan city in northern England. DESIGN: A quasi-experimental study comparing a group of older people before and after the introduction of an IC service. A quota sampling method was used to match the groups. SUBJECTS: Patients presenting as emergency admissions to two elderly care departments with falls, confusion, incontinence or immobility. INTERVENTION: a city-wide service in which a joint care management team (multi-agency, multi-disciplinary) assessed patient need and purchased support and rehabilitation from sector-based IC teams. OUTCOMES: Nottingham Extended Activities of Daily Living score, Barthel Index, Hospital Anxiety and Depression score, mortality, readmission to hospital, and new institutional care placement at 3, 6 and 12 months post-recruitment. RESULTS: There were 800 and 848 patients, respectively, in the control and intervention groups. Clinical outcomes, hospital and long-term care use were similar between the groups. Uptake of IC was lower than anticipated at 29%. An embedded case-control study comparing the 246 patients who received IC with a matched sample from the control group demonstrated similar clinical outcomes but increased hospital bed days used over 12 months (mean +8 days; 95% CI 3.1-13.0). CONCLUSION: This city-wide IC service was associated with similar clinical outcomes but did not achieve its strategic objectives of reducing long-term care and hospital use. PMID- 16267184 TI - Improving care for patients with dysphagia. AB - BACKGROUND: Early diagnosis and effective management of dysphagia reduce the incidence of pneumonia and improve quality of care and outcome. Dysphagic stroke patients rarely perceive that they have a swallowing problem, and thus carers have to take responsibility for following the safe swallow recommendations made by the Speech and Language Therapist (SLT). Published work and observations in our own Trust indicated that patients with dysphagia may be fed in a manner which places them at significant risk of aspiration, despite SLT advice for safe swallowing. OBJECTIVE: To determine compliance with swallowing recommendations in patients with dysphagia and to investigate the effectiveness of changes in practice in improving compliance. DESIGN: Sequential observational study before and after targeted intervention. SETTING: An acute general and teaching hospital in an inner city area. SUBJECTS: All patients with dysphagia on the caseload of the speech and language therapy department at the time of the study. METHODS: Observations were made on compliance with the recommendations of SLTs regarding consistency of fluids, dietary modifications, amount to be given at a single meal/drink, swallowing strategies, general safe swallow recommendations and whether supervision was required. A dysphagia link nurse programme was established, together with modification of an in-house training scheme, use of pre-thickened drinks and modification of swallowing advice sheets. The same observations were repeated after this intervention. RESULTS: Thirty-one patients were observed before and 54 after the intervention. There was improvement in compliance with the recommendations on consistency of fluids (48-64%, P<0.05), amount given (35-69%, P<0.05), adherence to safe swallow guidelines (51-90%, P<0.01) and use of supervision (35-67%, P<0.01). There were no significant differences in compliance with dietary modifications or swallowing strategies. Improvement in compliance was demonstrated in medical and geriatric wards and the stroke unit, but not in the surgical wards. Compliance with 'nil by mouth' instructions was 100% throughout. CONCLUSIONS: Relatively simple and low-cost measures, including an educational programme tailored to the needs of individual disciplines, proved effective in improving the compliance with advice on swallowing in patients with dysphagia. It is suggested that this approach may produce widespread benefit to patients across the NHS. PMID- 16267185 TI - Mobility disability in the middle-aged: cross-sectional associations in the English Longitudinal Study of Ageing. AB - BACKGROUND: Mobility (locomotor) disability is an early marker of disability progression, health care utilisation and institutionalisation in older people. Whether mobility disability has different causes in the middle-aged has received limited attention. OBJECTIVES: To examine associations of mobility disability with sociodemographic, behaviour and disease status and to contrast these with associations in older groups. DESIGN: Cross-sectional interview data from the 2002 English Longitudinal Study of Ageing. Mobility status based on reported difficulty walking a quarter of a mile (402 m). PARTICIPANTS: A total of 11,392 community-living respondents aged 50 years and over. RESULTS: In the middle-aged, 8% (95% CI 7-9%) of women and 9% (95% CI 8-11%) of men reported having much difficulty or being unable to walk a quarter of a mile, equating to 787,000 (95% CI 700,000-831,000) people in England. Factors which at least doubled odds of mobility disability in the middle-aged were chronic obstructive lung disease, angina, stroke, recently treated cancer, comorbidity, lower limb and back pain. Factors associated with mobility disability in older groups were similar. Thirty eight per cent of mobility disability in the middle-aged population was related to high levels of lower limb pain and 15% to high levels of back pain. CONCLUSIONS: Mobility disability in the middle-aged is relatively common. The associated conditions in the middle-aged are similar to those in older people. Lower limb and back pain make dominant population contributions to mobility disability. Prevention of later disability progression may require more attention being paid to mobility difficulties and its causes in the middle-aged. PMID- 16267186 TI - Enzymes of drug metabolism during delirium. AB - BACKGROUND: Delirium is common in ill medical patients. Several drugs and polypharmacy are recognised risk factors, yet little is known about drug metabolism in people with delirium. OBJECTIVE: The aim of this study was to investigate the activities of plasma esterases (drug metabolising enzymes) in delirium. DESIGN: This was a prospective study of delirium present at time of hospital admission (community acquired) or developing later (hospital acquired) in patients admitted as a medical emergency and aged 75 years or over. METHODS: Following informed consent or assent cognitive screening was completed on all patients on admission and every 48 hours subsequently. Delirium was diagnosed by Confusion Assessment Method and DSM IV criteria. Blood samples were taken on admission and at onset of delirium if this was later. Four plasma esterase assays were performed spectrophotometrically: acetylcholinesterase, aspirin esterase, benzoylcholinesterase, butyrylcholinesterase. RESULTS: 283 patients (71% of eligible) were recruited, with mean age 82.4 years and 59% female. 27% had community acquired delirium, 10% developed hospital acquired delirium, 63% never developed delirium. On admission the mean activities of all four esterase assays were statistically significantly lower in delirious than non delirious patients. There were no significant differences on admission in any plasma esterase activity between patients with hospital and community acquired delirium. In hospital mortality was associated with low plasma esterase activities on admission. CONCLUSION: Plasma esterase activities are suppressed during delirium. These data reinforce the need for extreme caution with drugs in this vulnerable population. PMID- 16267187 TI - Age-related differences in the areas of Broca and Wernicke using functional magnetic resonance imaging. AB - BACKGROUND: Previous studies have identified differences in the brain activation pattern between children and adults as well as between healthy subjects and patients using various imaging methods. Despite the increase of old people in the population, at present no study has applied a language task to investigate differences in adults. OBJECTIVE: We used a simple paradigm to elicit activation in young and old adults to investigate differences in the activation of the classical Broca and Wernicke areas. METHODS: Using functional magnetic resonance imaging, we applied a simple language paradigm to 10 right-handed subjects in each age group. Words and letter strings were presented separately with the instruction to decide for each pair if it represents a synonym (in case of words) or identical strings (in case of letters). The corresponding changes in the brain activation for words and letters were contrasted for each individual subject and compared by statistical parametric methods. RESULTS: For all subjects, the paradigm resulted in the activation of Broca and Wernicke areas in the left hemisphere only. Group analysis demonstrated a higher level of activation in Broca as well as in Wernicke areas for the young adults in comparison with the old adults. CONCLUSIONS: The applied, short paradigm provided consistent activation of the classical Broca and Wernicke areas in both age groups. These results demonstrate specific age-related differences in the processing of language stimuli during identical performance measures. PMID- 16267188 TI - Development and initial validation of the Falls Efficacy Scale-International (FES I). AB - BACKGROUND: There is a need for a measure of fear of falling that assesses both easy and difficult physical activities and social activities and is suitable for use in a range of languages and cultural contexts, permitting direct comparison between studies and populations in different countries and settings. OBJECTIVE: To develop a modified version of the Falls Efficacy Scale to satisfy this need, and to establish its psychometric properties, reliability, and concurrent validity (i.e. that it demonstrates the expected relationship with age, falls history and falls risk factors). DESIGN: Cross-sectional survey. SETTING: Community sample. METHOD: 704 people aged between 60 and 95 years completed The Falls Efficacy Scale-International (FES-I) either in postal self-completion format or by structured interview. RESULTS: The FES-I had excellent internal and test-retest reliability (Cronbach's alpha=0.96, ICC=0.96). Factor analysis suggested a unitary underlying factor, with two dimensions assessing concern about less demanding physical activities mainly in the home, and concern about more demanding physical activities mainly outside the home. The FES-I had slightly better power than the original FES items to discriminate differences in concern about falling between groups differentiated by sex, age, occupation, falls in the past year, and falls risk factors (chronic illness, taking multiple or psychoactive medications, dizziness). CONCLUSIONS: The FES-I has close continuity with the best existing measure of fear of falling, excellent psychometric properties, and assesses concerns relating to basic and more demanding activities, both physical and social. Further research is required to confirm cross-cultural and predictive validity. PMID- 16267189 TI - Relations between undernutrition and nosocomial infections in elderly patients. AB - BACKGROUND: Hospital-acquired infections and malnutrition are of major concern in public health in elderly patients. However, the interactions between these two entities are not well established. OBJECTIVES: To determine the incidence of nosocomial infections (NI) and its association with malnutrition. SUBJECTS: 185 hospitalised older adults aged 81.6+/-0.6 years old were nutritionally assessed on admission by measurement of anthropometric variables, serum nutritional proteins and evaluation of dietary intake. During hospitalisation, patients' progress was closely monitored, particularly for the detection of nosocomial infections. RESULTS: The incidence rate of NI was 59% and the global infection rate was 7.6/1,000 bed days. The most common infection site was the urinary tract (n=63). The nutritional status of the population was studied by comparing three groups defined according to the absence (group I, n=116), presence of one infection (group II, n=38) or presence of more than one infection (group III, n=31). All but one anthropometric parameters varied among the three groups. Total energy intake also varied among the three groups. The group I had higher daily nutrient intake than the other two groups (respectively P=0.004 and P<0.0001). Albumin, transthyretin, and C-reactive protein levels differed significantly among the three groups (respectively P<0.0001, P<0.0001 and P=0.0003). Age, energy intake, length of hospital stay and the presence of a urinary catheter were independent risk factors of nosocomial infection. CONCLUSION: Our findings show that patients with multiple NI were older, showed an altered nutritional status, a prolonged recovery, more frequently had urinary catheters and more discharge placement. PMID- 16267190 TI - Medication management at home: medication-related risk factors associated with poor health outcomes. AB - BACKGROUND: Some patients may have medication-related risk factors only identified by home visits, but the extent to which those risk factors are associated with poor health outcomes remains unclear. OBJECTIVE: To determine the association between medication-related risk factors and poor patient health outcomes from observations in the patients' homes. DESIGN: Cross-sectional study. SETTING: Patients' homes. SUBJECTS: 204 general practice patients living in their own homes and at risk of medication-related poor health outcomes. METHODS: Medications and medication-related risk factors were identified in the patients' homes by community pharmacists and general practitioners (GPs). The medication related risk factors were examined as determinants of patients' self-reported health related quality of life (SF-36) and their medication use, as well as physicians' impression of patient adverse drug events and health status. RESULTS: Key medication-related risk factors associated with poor health outcomes included: Lack of any medication administration routine, therapeutic duplication, hoarding, confusion between generic and trade names, multiple prescribers, discontinued medication repeats retained and multiple storage locations. Older age and female gender were associated with some poorer health outcomes. In addition, expired medication and poor adherence were also associated with poor health outcomes, however, not independently. CONCLUSION: The findings support the theory that polypharmacy and medication-related risk factors as a result of polypharmacy are correlated to poor health outcomes. PMID- 16267191 TI - Intermittent intramuscular clodronate therapy: a valuable option for older osteoporotic women. PMID- 16267193 TI - Age-related differences in smoking cessation advice and support given to patients hospitalised with smoking-related illness. PMID- 16267192 TI - Tailored group exercise (Falls Management Exercise -- FaME) reduces falls in community-dwelling older frequent fallers (an RCT). PMID- 16267194 TI - In-hospital hip fractures: prevalence, risk factors and outcome. PMID- 16267195 TI - An unusual cause of chest pain. AB - Flail chest is an uncommon condition that generally arises due to a significant impact to the chest, resulting in multiple fractures of the anterior and posterior ribs. This force may be much less if the bones are weakened for any reason, in osteoporosis or myeloma for instance. We describe a case of flail segment that arose secondary to a large sternal abscess resulting from methicillin-resistant Staphylococcus aureus bacteraemia. PMID- 16267196 TI - Collapse in a 79-year-old: a rare case of amyloid tumour of the pelvis. AB - A 79-year-old man presented to accident and emergency with collapse, unable to bear weight on his left leg. Computed tomography revealed a large isolated lesion (28 x 12 x 8 cm) extending from the pelvis into the abdomen, affecting the left lumbrosacral nerves. Further investigations showed that the mass contained amyloid protein. With no evidence of systemic amyloidosis or malignancy a diagnosis of amyloidoma/amyloid tumour was made. This is the largest amyloid tumour reported in the literature to date. There is limited but conflicting evidence regarding the pathophysiology, management and prognosis of amyloidoma. Clearly amyloidomas are rare, but patients can present acutely and may have a poor prognosis, especially when the tumour is of considerable size. PMID- 16267197 TI - Buried Bumper Syndrome complicated by intra-abdominal sepsis. AB - There is growing evidence that enteral feeding tubes are associated with increased mortality and complication rates in patients with advanced dementia. Buried Bumper Syndrome is an uncommon, but well documented late complication of PEG placement. Our case report reinforces this recognised risk of PEG feeding in an elderly, cognitively impaired patient. PMID- 16267198 TI - Trench foot following a collapse: assessment of the feet is essential in the elderly. AB - Elderly patients commonly present to hospital following a collapse and period of distressing immobilisation on the floor. We present a case of bilateral trench foot in such a patient with no prior peripheral vascular disease. Examination of the feet is mandatory for early detection of this rare condition in the collapsed elderly patient. PMID- 16267199 TI - Primary malignant melanoma of the nose: a rare cause of epistaxis in the elderly. AB - We report the case of a primary nasal malignant melanoma in an elderly woman, illustrating the dilemmas involved in clinical decision making with non-specific symptoms, and the treatment of cancer in elderly patients. We also discuss the incidence, clinical presentation, diagnosis, principles of management and outcome. PMID- 16267200 TI - Inter-rater reliability of the Barthel Index. PMID- 16267201 TI - Methods to correct placement of a nasogastric tube: beware of the pitfalls. PMID- 16267202 TI - Musical abilities may outlast other faculties in advanced dementia. PMID- 16267203 TI - Response to: Hypodermoclysis--a victim of historical prejudice. PMID- 16267204 TI - Traumatic events and recall. PMID- 16267207 TI - Novel and specific inhibitors of a poxvirus type I topoisomerase. AB - Vaccinia DNA topoisomerase (vTopo) is a prototypic pox virus family topoisomerase that shares extensive structural and mechanistic properties with the human type IB enzyme (hTopo) and is important for viral replication. Despite their far reaching similarities, vTopo and hTopo have surprisingly distinct pharmacological properties. To further exploit these differences, we have developed recently the first high-throughput screen for vTopo, which has allowed rapid screening of a 1990-member small-molecule library for inhibitors. Using this approach, 21 compounds were identified with IC(90) values less than 10 muM, and 19 of these were also found to inhibit DNA supercoil relaxation by vTopo. Four of the most potent compounds were completely characterized and are structurally novel topo I inhibitors with efficacies at nanomolar concentrations. These inhibitors were highly specific for vTopo, showing no inhibition of the human enzyme even at 500- to 2000-fold greater concentrations. We describe a battery of efficient experiments to characterize the unique mechanisms of these vTopo inhibitors and discuss the surprising promiscuity of this enzyme to inhibition by structurally diverse small molecules. PMID- 16267208 TI - Activation and potentiation of interferon-gamma signaling by 3,3' diindolylmethane in MCF-7 breast cancer cells. AB - 3,3'-Diindolylmethane (DIM), a natural autolytic product in plants of the Brassica genus, including broccoli, cauliflower, and Brussels sprouts, exhibits promising cancer protective activities, especially against mammary neoplasia in animal models. We observed previously that DIM induced a G(1) cell-cycle arrest and strong induction of cell-cycle inhibitor p21 expression and promoter activity in both estrogen-responsive and -independent breast cancer cell lines. We showed recently that DIM up-regulates the expression of interferon gamma (IFNgamma) in human MCF-7 breast cancer cells. This novel effect may contribute to the anticancer effects of DIM because IFNgamma plays an important role in preventing the development of primary and transplanted tumors. In this study, we observed that DIM activated the IFNgamma signaling pathway in human breast cancer cells. DIM activated the expression of the IFNgamma receptor (IFNGR1) and IFNgamma responsive genes p56- and p69-oligoadenylate synthase (OAS). In cotreatments with IFNgamma, DIM produced an additive activation of endogenous p69-OAS and of an OAS Luc reporter and a synergistic activation of a GAS-Luc reporter. DIM synergistically augmented the IFNgamma induced phosphorylation of signal transducer and activator of transcription factor 1, further evidence of DIM activation of the IFNgamma pathway. DIM and IFNgamma produced an additive inhibition of cell proliferation and a synergistic increase in levels of major histocompatibility complex class-1 (MHC-1) expression, accompanied by increased levels of mRNAs of MHC-1-associated proteins and transporters. These results reveal novel immune activating and potentiating activities of DIM in human tumor cells that may contribute to the established effectiveness of this dietary indole against various tumors types. PMID- 16267209 TI - Four novel tarantula toxins as selective modulators of voltage-gated sodium channel subtypes. AB - Four novel peptide toxins that act on voltage-gated sodium channels have been isolated from tarantula venoms: ceratotoxins 1, 2, and 3 (CcoTx1, CcoTx2, and CcoTx3) from Ceratogyrus cornuatus and phrixotoxin 3 (PaurTx3) from Phrixotrichus auratus. The pharmacological profiles of these new toxins were characterized by electrophysiological measurements on six cloned voltage-gated sodium channel subtypes expressed in Xenopus laevis oocytes (Na(v)1.1/beta(1), Na(v)1.2/beta(1), Na(v)1.3/beta(1), Na(v)1.4/beta(1), Na(v)1.5/beta(1), and Na(v)1.8/beta(1)). These novel toxins modulate voltage-gated sodium channels with properties similar to those of typical gating-modifier toxins, both by causing a depolarizing shift in gating kinetics and by blocking the inward component of the sodium current. PaurTx3 is one of the most potent peptide modulators of voltage-gated sodium channels described thus far from spider venom, modulating Na(v)1.2 with an IC(50) value of 0.6 +/- 0.1 nM. CcoTx1 and CcoTx2, differing by only one amino acid, are potent modulators of different voltage-gated sodium channel subtypes from the central nervous system, except for Na(v)1.3, which is only affected by CcoTx2. The potency of CcoTx3 is lower, although this toxin seems to be more selective for the tetrodotoxin-resistant channel subtype Na(v)1.5/beta(1) (IC(50) = 447 +/- 32 nM). In addition to these results, molecular modeling indicates that subtle differences in toxin surfaces may relate to their different pharmacological profiles. Furthermore, an evolutionary trace analysis of these toxins and other structurally related three-disulfide spider toxins provides clues for the exploration of toxin-channel interaction and future structure-function research. PMID- 16267210 TI - Leptin in pregnancy: an update. AB - Leptin influences satiety, adiposity, and metabolism and is associated with mechanisms regulating puberty onset, fertility, and pregnancy in various species. Maternal hyperleptinemia is a hallmark of mammalian pregnancy, although both the roles of leptin and the mechanisms regulating its synthesis appear to be taxa specific. In pregnant humans and nonhuman primates, leptin is produced by both maternal and fetal adipose tissues, as well as by the placental trophoblast. Specific receptors in the uterine endometrium, trophoblast, and fetus facilitate direct effects of the polypeptide on implantation, placental endocrine function, and conceptus development. A soluble isoform of the receptor may be responsible for inducing maternal leptin resistance during pregnancy and/or may facilitate the transplacental passage of leptin for the purpose of directly regulating fetal development. The steroid hormones are linked to the regulation of leptin and the leptin receptor and probably interact with other pregnancy-specific, serum-borne factors to regulate leptin dynamics during pregnancy. In addition to its effects on normal conceptus development, leptin is linked to mechanisms affecting a diverse array of pregnancy-specific pathologies that include preeclampsia, gestational diabetes, and intrauterine growth restriction. Association with these anomalies and with mechanisms pointing to a fetal origin for a range of conditions affecting the individual's health in adult life, such as obesity, diabetes mellitus, and cardiovascular disease, reiterate the need for continued research dedicated to elucidating leptin's roles and regulation throughout gestation. PMID- 16267211 TI - Extensive S aureus spinal epidural empyema. PMID- 16267212 TI - Morphine-induced mu-opioid receptor internalization: a paradox solved in neurons. PMID- 16267214 TI - Neuronal differentiation in the adult hippocampus recapitulates embryonic development. AB - In the adult hippocampus and olfactory bulb, neural progenitor cells generate neurons that functionally integrate into the existing circuits. To understand how neuronal differentiation occurs in the adult hippocampus, we labeled dividing progenitor cells with a retrovirus expressing green fluorescent protein and studied the morphological and functional properties of their neuronal progeny over the following weeks. During the first week neurons had an irregular shape and immature spikes and were synaptically silent. Slow GABAergic synaptic inputs first appeared during the second week, when neurons exhibited spineless dendrites and migrated into the granule cell layer. In contrast, glutamatergic afferents were detected by the fourth week in neurons displaying mature excitability and morphology. Interestingly, fast GABAergic responses were the latest to appear. It is striking that neuronal maturation in the adult hippocampus follows a precise sequence of connectivity (silent --> slow GABA --> glutamate --> fast GABA) that resembles hippocampal development. We conclude that, unlike what is observed in the olfactory bulb, the hippocampus maintains the same developmental rules for neuronal integration through adulthood. PMID- 16267213 TI - Olig transcription factors are expressed in oligodendrocyte and neuronal cells in human fetal CNS. AB - The transcription factors Olig1 and Olig2 are closely associated with the development of oligodendrocyte (OL) lineage in the vertebrate nervous system, but little is known about their role in the human developing CNS. To test the hypothesis that they contribute to initial OL specification in humans, we studied the expression of Olig1 and Olig2 in human fetuses at 5-24 gestational weeks (GW). Both transcription factors were present in well outlined regions of the ventral neuroepithelium at 5 GW, several weeks before oligodendrogenesis. Spatial differences in the expression of Olig1 and Olig2 along the neuronal axis suggest that they specify different subpopulations of progenitor cells. Olig1 was distributed rostrally, from the basal forebrain to the hindbrain, whereas Olig2 was also found in the ventral spinal cord. Furthermore, at 5 GW, Olig1 was coexpressed with vimentin, and Olig2 was coexpressed with a neuronal marker, microtubule-associated protein 2. With the progression of development at 15 GW, both proteins were present throughout the spinal cord and the ventricular subventricular zone of the ganglionic eminences, whereas at midgestation (20 GW), they were also expressed in the telencephalic proliferative zones and the emerging white matter. Double-labeling studies revealed that early OL progenitor cells and radial glia expressed Olig1, whereas Olig2 was localized predominantly in mature OLs and a subset of neural progenitor cells and mature neurons. Thus, Olig1 and Olig2 transcription factors in the human CNS are important not only for differentiation of the OL lineage, but they may also have a role in neural cell specification. PMID- 16267215 TI - Input to V2 thin stripes arises from V1 cytochrome oxidase patches. AB - Parallel streams from the primary visual cortex (V1) to the second visual area (V2) are thought to mediate different aspects of visual perception in primates. One hypothesis is that the projection from cytochrome oxidase patches to thin stripes is responsible for color, whereas a separate pathway from interpatches to pale stripes mediates form. Recently, the notion of segregated pathways has been challenged by a report showing that patches and interpatches project equally to thin stripes. We made injections of a retrograde tracer, cholera toxin-B (CTB Au), into macaque V2 thin stripes and counted the number of labeled cells in patches versus interpatches in layer 2/3. Analysis of eight thin-stripe injections showed that a mean of 81% of labeled cells were located in patches (defined as 33% of the surface area of V1). This result confirms that the projection to thin stripes arises predominately from patches. To assess the segregation of patch and interpatch projections, we injected CTB-Au in a pale stripe and horseradish peroxidase in an adjacent thin stripe. In both successful cases, interdigitated fields of labeled cells were present in V1. Less than 1% of cells were double-labeled, indicating that the populations of cells supplying thin stripes and pale stripes are quite independent. This finding means that different signals are likely conveyed by patches and interpatches to V2. PMID- 16267217 TI - Neural coupling binds visual tokens to moving stimuli. AB - Spatially separated visual objects that appear in alternating sequence can be perceived as a single moving object. This phenomenon of apparent motion enables us to perceive sequentially presented images as a motion picture. How does the visual system bind together single visual tokens to one moving object? Here we report a series of experiments investigating apparent motion with electroencephalographic recordings showing that gamma-band oscillatory coupling in the visual cortex is crucial for this phenomenon. We used an ambiguous stimulus that could be perceived as moving either vertically or horizontally. Because visual information from the right and left visual hemifields is routed to the contralateral early visual cortex, in the case of perceived horizontal movement, information from both hemispheres has to be integrated. In accord with this assumption, our data show stronger oscillatory coupling between right and left visual cortices during perception of horizontal motion compared with vertical motion. PMID- 16267216 TI - Tumor necrosis factor-like weak inducer of apoptosis increases the permeability of the neurovascular unit through nuclear factor-kappa B pathway activation. AB - Tumor necrosis factor-like weak inducer of apoptosis (TWEAK) is a member of the tumor necrosis factor superfamily. TWEAK acts on responsive cells via binding to a small cell-surface receptor named fibroblast growth factor-inducible-14 (Fn14). TWEAK can stimulate numerous cellular responses including cell proliferation, migration, and proinflammatory molecule production. The present study investigated whether TWEAK plays a role in the regulation of the permeability of the neurovascular unit (NVU). We found that intracerebral injection of TWEAK in wild-type mice induces activation of the nuclear factor-kappaB (NF-kappaB) pathway and matrix metalloproteinase-9 (MMP-9) expression in the brain with resultant disruption in the structure of the NVU and increase in the permeability of the blood-brain barrier (BBB). TWEAK did not increase MMP-9 activity or BBB permeability when injected into mice genetically deficient in the NF-kappaB family member p50. Furthermore, we report that inhibition of TWEAK activity during cerebral ischemia with an Fn14-Fc decoy receptor results in significant preservation of the integrity of the NVU with attenuation of cerebral ischemia induced increase in the permeability of the BBB. We conclude that the cytokine TWEAK plays a role in the disruption of the structure and permeability of the NVU during physiological and pathological conditions. PMID- 16267218 TI - cAMP response element-binding protein regulates differentiation and survival of newborn neurons in the olfactory bulb. AB - The transcription factor cAMP response element-binding protein (CREB) is involved in multiple aspects of neuronal development and plasticity. Here, we demonstrate that CREB regulates specific phases of adult neurogenesis in the subventricular zone/olfactory bulb (SVZ/OB) system. Combining immunohistochemistry with bromodeoxyuridine treatments, cell tracer injections, cell transplants, and quantitative analyses, we show that although CREB is expressed by the SVZ neuroblasts throughout the neurogenic process, its phosphorylation is transient and parallels neuronal differentiation, increasing during the late phase of tangential migration and decreasing after dendrite elongation and spine formation. In vitro, inhibition of CREB function impairs morphological differentiation of SVZ-derived neuroblasts. Transgenic mice lacking CREB, in a null CREM genetic background, show reduced survival of newborn neurons in the OB. This finding is further supported by peripheral afferent denervation experiments resulting in downregulation of CREB phosphorylation in neuroblasts, the survival of which appears heavily impaired. Together, these findings provide evidence that CREB regulates differentiation and survival of newborn neurons in the OB. PMID- 16267219 TI - Molecular control of spinal accessory motor neuron/axon development in the mouse spinal cord. AB - Within the developing vertebrate spinal cord, motor neuron subtypes are distinguished by the settling positions of their cell bodies, patterns of gene expression, and the paths their axons follow to exit the CNS. The inclusive set of cues required to guide a given motor axon subtype from cell body to target has yet to be identified, in any species. This is attributable, in part, to the unavailability of markers that demarcate the complete trajectory followed by a specific class of spinal motor axons. Most spinal motor neurons extend axons out of the CNS through ventral exit points. In contrast, spinal accessory motor neurons (SACMNs) project dorsally directed axons through lateral exit points (LEPs), and these axons assemble into the spinal accessory nerve (SAN). Here we show that an antibody against BEN/ALCAM/SC1/DM-GRASP/MuSC selectively labels mouse SACMNs and can be used to trace the pathfinding of SACMN axons. We use this marker, together with a battery of transcription factor-deficient or guidance cue/receptor-deficient mice to identify molecules required for distinct stages of SACMN development. Specifically, we find that Gli2 is required for the initial extension of axons from SACMN cell bodies, and that netrin-1 and its receptor Dcc are required for the proper dorsal migration of these cells and the dorsally directed extension of SACMN axons toward the LEPs. Furthermore, in the absence of the transcription factor Nkx2.9, SACMN axons fail to exit the CNS. Together, these findings suggest molecular mechanisms that are likely to regulate key steps in SACMN development. PMID- 16267220 TI - Breakdown of long-range temporal correlations in theta oscillations in patients with major depressive disorder. AB - Neuroimaging has revealed robust large-scale patterns of high neuronal activity in the human brain in the classical eyes-closed wakeful rest condition, pointing to the presence of a baseline of sustained endogenous processing in the absence of stimulus-driven neuronal activity. This baseline state has been shown to differ in major depressive disorder. More recently, several studies have documented that despite having a complex temporal structure, baseline oscillatory activity is characterized by persistent autocorrelations for tens of seconds that are highly replicable within and across subjects. The functional significance of these long-range temporal correlations has remained unknown. We recorded neuromagnetic activity in patients with a major depressive disorder and in healthy control subjects during eyes-closed wakeful rest and quantified the long range temporal correlations in the amplitude fluctuations of different frequency bands. We found that temporal correlations in the theta-frequency band (3-7 Hz) were almost absent in the 5-100 s time range in the patients but prominent in the control subjects. The magnitude of temporal correlations over the left temporocentral region predicted the severity of depression in the patients. These data indicate that long-range temporal correlations in theta oscillations are a salient characteristic of the healthy human brain and may have diagnostic potential in psychiatric disorders. We propose a link between the abnormal temporal structure of theta oscillations in the depressive patients and the systems-level impairments of limbic-cortical networks that have been identified in recent anatomical and functional studies of patients with major depressive disorder. PMID- 16267221 TI - Declarative memory, awareness, and transitive inference. AB - A characteristic usually attributed to declarative memory is that what is learned is accessible to awareness. Recently, the relationship between awareness and declarative (hippocampus-dependent) memory has been questioned on the basis of findings from transitive inference tasks. In transitive inference, participants are first trained on overlapping pairs of items (e.g., A+B-, B+C-, C+D-, and D+E , where + and - indicate correct and incorrect choices). Later, participants who choose B over D when presented with the novel pair BD are said to demonstrate transitive inference. The ability to exhibit transitive inference is thought to depend on the fact that participants have represented the stimulus elements hierarchically (i.e., A>B>C>D>E). We found that performance on five-item and six item transitive inference tasks was closely related to awareness of the hierarchical relationship among the elements of the training pairs. Participants who were aware of the hierarchy performed near 100% correct on all tests of transitivity, but participants who were unaware of the hierarchy performed poorly (e.g., on transitive pair BD in the five-item problem; on transitive pairs BD, BE, and CE in the six-item problem). When the five-item task was administered to memory-impaired patients with damage thought to be limited to the hippocampal region, the patients were impaired at learning the training pairs. All patients were unaware of the hierarchy and, like unaware controls, performed poorly on the BD pair. The findings indicate that awareness is critical for robust performance on tests of transitive inference and support the view that awareness of what is learned is a fundamental characteristic of declarative memory. PMID- 16267222 TI - A Drosophila KCNQ channel essential for early embryonic development. AB - The mammalian voltage-dependent KCNQ channels are responsible for distinct types of native potassium currents and are associated with several human diseases. We cloned a novel Drosophila KCNQ channel (dKCNQ) based on its sequence homology to the mammalian genes. When expressed in Chinese hamster ovary cells, dKCNQ gives rise to a slowly activating and slowly deactivating current that activates in the subthreshold voltage range. Like the M-current produced by mammalian KCNQ channels, dKCNQ current is sensitive to the KCNQ-specific blocker linopirdine and is suppressed by activation of a muscarinic receptor. dKCNQ is also similar to the mammalian channels in that it binds calmodulin (CaM), and CaM binding is necessary to produce functional currents. In situ hybridization analysis demonstrates that dKCNQ mRNA is present in brain cortical neurons, the cardia (proventriculus), and the nurse cells and oocytes of the ovary. We generated mutant flies with deletions in the genomic sequence of dKCNQ. Embryos produced by homozygous deletion females exhibit disorganized nuclei and fail to hatch, suggesting strongly that a maternal contribution of dKCNQ protein and/or mRNA is essential for early embryonic development. PMID- 16267223 TI - Glucagon-expressing neurons within the retina regulate the proliferation of neural progenitors in the circumferential marginal zone of the avian eye. AB - Glucagon-expressing retinal amacrine cells have been implicated in regulating postnatal ocular growth. Furthermore, experimentally accelerated rates of ocular growth increase the number of neurons added to the peripheral edge of the retina. Accordingly, we assayed whether glucagon-expressing neurons within the retina regulate the proliferation of progenitors in the circumferential marginal zone (CMZ) of the postnatal chicken eye. We found that glucagon-containing neurites are heavily clustered within the CMZ at the peripheral edge of the retina. Many of these neurites originate from a cell type that is distinct from other types of retinal neurons, which we termed large glucagon-expressing neurons (LGENs). The LGENs are immunoreactive for glucagon and glucagon-like peptide 1 (GLP1), have a unipolar morphology, produce an axon that projects into the CMZ, and are found only in ventral regions of the retina. In dorsal regions of the retina, a smaller version of the LGENs densely ramifies neurites in the CMZ. Intraocular injections of glucagon or GLP1 suppressed the proliferation of progenitors in the CMZ, whereas a glucagon-receptor antagonist promoted proliferation. In addition, we found that glucagon, GLP1, and glucagon antagonist influenced the number of progenitors in the CMZ. We conclude that the LGENs may convey visual information to the CMZ to control the addition of new cells to the edge of the retina. We propose that glucagon/GLP1 released from LGENs acts in opposition to insulin (or insulin-like growth factor) to regulate precisely the proliferation of retinal progenitors in the CMZ. PMID- 16267224 TI - Extensive cortical rewiring after brain injury. AB - Previously, we showed that the ventral premotor cortex (PMv) underwent neurophysiological remodeling after injury to the primary motor cortex (M1). In the present study, we examined cortical connections of PMv after such lesions. The neuroanatomical tract tracer biotinylated dextran amine was injected into the PMv hand area at least 5 months after ischemic injury to the M1 hand area. Comparison of labeling patterns between experimental and control animals demonstrated extensive proliferation of novel PMv terminal fields and the appearance of retrogradely labeled cell bodies within area 1/2 of the primary somatosensory cortex after M1 injury. Furthermore, evidence was found for alterations in the trajectory of PMv intracortical axons near the site of the lesion. The results suggest that M1 injury results in axonal sprouting near the ischemic injury and the establishment of novel connections within a distant target. These results support the hypothesis that, after a cortical injury, such as occurs after stroke, cortical areas distant from the injury undergo major neuroanatomical reorganization. Our results reveal an extraordinary anatomical rewiring capacity in the adult CNS after injury that may potentially play a role in recovery. PMID- 16267225 TI - Deletion of the prostaglandin E2 EP2 receptor reduces oxidative damage and amyloid burden in a model of Alzheimer's disease. AB - Epidemiological studies demonstrate that chronic use of nonsteroidal anti inflammatory drugs (NSAIDs) in normal aging populations reduces the risk of developing Alzheimer's disease (AD). NSAIDs inhibit the enzymatic activity of cyclooxygenase-1 (COX-1) and inducible COX-2, which catalyze the first committed step in the synthesis of prostaglandins. These studies implicate COX-mediated inflammation as an early and potentially reversible preclinical event; however, the mechanism by which COX activity promotes development of AD has not been determined. Recent studies implicate the prostaglandin E2 (PGE2) E prostanoid subtype 2 (EP2) receptor in the development of the innate immune response in brain. Here, we report that deletion of the PGE2 EP2 receptor in the APPSwe PS1DeltaE9 model of familial AD results in marked reductions in lipid peroxidation in aging mice. This reduction in oxidative stress is associated with significant decreases in levels of amyloid-beta (Abeta) 40 and 42 peptides and amyloid deposition. Aged APPSwe-PS1DeltaE9 mice lacking the EP2 receptor harbor lower levels of beta C-terminal fragments, the product of beta-site APP cleaving enzyme (BACE1) processing of amyloid precursor protein. Increases in BACE1 processing have been demonstrated in models of aging and AD and after oxidative stress. Our results indicate that PGE2 signaling via the EP2 receptor promotes age-dependent oxidative damage and increased Abeta peptide burden in this model of AD, possibly via effects on BACE1 activity. Our findings identify EP2 receptor signaling as a novel proinflammatory and proamyloidogenic pathway in this model of AD, and suggest a rationale for development of therapeutics targeting the EP2 receptor in neuroinflammatory diseases such as AD. PMID- 16267226 TI - N-terminal insertion and C-terminal ankyrin-like repeats of alpha-latrotoxin are critical for Ca2+-dependent exocytosis. AB - Alpha-latrotoxin, a potent stimulator of exocytosis from neurons and neuroendocrine cells, has been studied intensively, but the mechanisms of its actions are poorly understood. Here, we developed a new method to generate active recombinant alpha-latrotoxin and conducted a structure/function analysis of the toxin in stimulating Ca2+-dependent exocytosis. alpha-Latrotoxin consists of a conserved N-terminal domain and C-terminal ankyrin-like repeats. After cleavage of an N-terminally fused purification tag of glutathione S-transferase (GST), the recombinant toxin strongly stimulated exocytosis, whereas the GST-fused toxin was much less potent. The GST-fused toxin bound to the receptors [neurexin 1alpha; CL1 (CIRL/latrophilin 1)] as efficiently as did the GST-cleaved toxin but was much less effective in inserting into the plasma membrane and inducing cation conductance. The toxin with deletion of the last two ankyrin-like repeats still bound the receptors but could neither stimulate exocytosis nor induce cation conductance efficiently. The abilities of the mutated toxins to stimulate exocytosis correlated well with their abilities to induce cation conductance, but not their binding to the receptors. Our results indicate that (1) C-terminal ankyrin-like repeats and a free (unfused) N terminus are both required for the toxin to form pores, which is essential for Ca2+-dependent exocytosis, and (2) receptor binding alone is not sufficient to stimulate Ca2+-dependent exocytosis. PMID- 16267227 TI - Antidepressants and antipsychotic drugs colocalize with 5-HT3 receptors in raft like domains. AB - Despite different chemical structure and pharmacodynamic signaling pathways, a variety of antidepressants and antipsychotics inhibit ion fluxes through 5-HT3 receptors in a noncompetitive manner with the exception of the known competitive antagonists mirtazapine and clozapine. To further investigate the mechanisms underlying the noncompetitive inhibition of the serotonin-evoked cation current, we quantified the concentrations of different types of antidepressants and antipsychotics in fractions of sucrose flotation gradients isolated from HEK293 (human embryonic kidney 293) cells stably transfected with the 5-HT3A receptor and of N1E-115 neuroblastoma cells in relation to the localization of the 5-HT3 receptor protein within the cell membrane. Western blots revealed a localization of the 5-HT3 receptor protein exclusively in the low buoyant density (LBD) fractions compatible with a localization within raft-like domains. Also, the antidepressants desipramine, fluoxetine, and reboxetine and the antipsychotics fluphenazine, haloperidol, and clozapine were markedly enriched in LBD fractions, whereas no accumulation occurs for mirtazapine, carbamazepine, moclobemide, and risperidone. The concentrations of psychopharmacological drugs within LBD fractions was strongly associated with their inhibitory potency against serotonin induced cation currents. The noncompetitive antagonism of antidepressants at the 5-HT3 receptor was not conferred by an enhancement of receptor internalization as shown by immunofluorescence studies, assessment of receptor density in clathrin coated vesicles, and electrophysiological recordings after coexpression of a dominant-negative mutant of dynamin I, which inhibits receptor internalization. In conclusion, enrichment of antidepressants and antipsychotics in raft-like domains within the cell membrane appears to be crucial for their antagonistic effects at ligand-gated ion channels such as 5-HT3 receptors. PMID- 16267228 TI - Neuronal computation of disparity in V1 limits temporal resolution for detecting disparity modulation. AB - The human ability to detect modulation of binocular disparity over time is poor compared with detection of luminance modulation. We examined the physiological origin of this limitation by analyzing neuronal responses to temporal modulation of binocular disparity in striate cortex of awake monkeys. When neurons were presented with random-dot stereograms in which disparity varied sinusoidally over time, their responses modulated at the stimulus temporal frequency, with little change in mean firing rate. We calculated modulation amplitude as a function of temporal frequency and compared this with the psychophysical performance of four human observers. Neuronal and psychophysical functions showed similar peak frequencies (2 Hz) and comparable high-cut frequencies (10 and 5.5 Hz, respectively). Thus, V1 (primary visual cortex) neurons appear to limit psychophysical performance. The temporal resolution of the same neurons for contrast modulation was approximately 2.5 times greater, which parallels the superior psychophysical performance for contrast. There is a simple mathematical explanation for this difference: it results from calculating cross-correlation between temporally broadband monocular images that are bandpass filtered before measuring correlation. The limit on temporal resolution is a direct consequence of the binocular energy model that adds to the list of properties of human stereoscopic performance that are explained by this simple model of disparity encoding in V1: the same neurons can account for the performance of psychophysical tasks that result in either high (contrast) or low (disparity) temporal resolution. Because this principle holds whenever a broadband input is bandpass filtered before computing correlation, it may limit the resolution of other neuronal systems. PMID- 16267229 TI - Nigrostriatal dysfunction in familial Alzheimer's disease-linked APPswe/PS1DeltaE9 transgenic mice. AB - Alzheimer's disease (AD) is often accompanied by extrapyramidal signs attributed to nigrostriatal dysfunction. The association between amyloid deposition and nigrostriatal degeneration is essentially unknown. We showed previously that the striatum and the substantia nigra of transgenic mice harboring familial AD (FAD) linked APPswe/PS1DeltaE9 mutants exhibit morphological alterations accompanied by amyloid-beta (Abeta) deposition (Perez et al., 2004). In the present study, we further investigated the interaction between Abeta deposition and dopaminergic nigrostriatal dysfunction, by correlating morphological and biochemical changes in the nigrostriatal pathway with amyloid deposition pathology in the brains of 3 to 17-month-old APPswe/PS1DeltaE9 transgenic mice and age-matched wild-type controls. We show that Abeta deposition is pronounced in the striatum of APPswe/PS1DeltaE9 mice at 6 months of age, and the extent of deposition increases in an age-dependent manner. Tyrosine hydroxylase (TH)-positive dystrophic neurites with rosette or grape-like cluster disposition are observed adjacent to Abeta plaques and display multilaminar, multivesicular, and dense-core bodies as well as mitochondria. In addition, an age-dependent increase of TH protein levels are shown in nigral cells in these mutant mice. Using HPLC analysis, we found a reduction in the dopamine metabolite DOPAC in the striatum of these mice. These findings show a close association between amyloid deposition and nigrostriatal pathology and suggest that altered FAD-linked amyloid metabolism impairs, at least in part, the function of dopaminergic neurons. PMID- 16267231 TI - Functional specialization in the human medial temporal lobe. AB - Investigations of memory in rats and nonhuman primates have demonstrated functional specialization within the medial temporal lobe (MTL), a set of heavily interconnected structures including the hippocampal formation and underlying entorhinal, perirhinal, and parahippocampal cortices. Most studies in humans, however, especially in patients with brain damage, suggest that the human MTL is a unitary memory system supporting all types of declarative memory, our conscious memory for facts and events. To resolve this discrepancy, amnesic patients with either selective hippocampal damage or more extensive MTL damage were tested on variations of an object discrimination task adapted from the nonhuman primate literature. Although both groups were equally impaired on standard recall-based memory tasks, they exhibited different profiles of performance on the object discrimination test, arguing against a unitary view of MTL function. Cases with selective hippocampal damage performed normally, whereas individuals with broader MTL lesions were impaired. Furthermore, deficits in this latter group were related not to the number of discriminations to be learned and remembered, but to the degree of "feature ambiguity," a property of visual discriminations that can emerge when features are part of both rewarded and unrewarded stimuli. These findings resolve contradictions between published studies in humans and animals and introduce a new way of characterizing the impairments that arise after damage to the MTL. PMID- 16267230 TI - Control of spontaneous firing patterns by the selective coupling of calcium currents to calcium-activated potassium currents in striatal cholinergic interneurons. AB - The spontaneous firing patterns of striatal cholinergic interneurons are sculpted by potassium currents that give rise to prominent afterhyperpolarizations (AHPs). Large-conductance calcium-activated potassium (BK) channel currents contribute to action potential (AP) repolarization; small-conductance calcium-activated potassium channel currents generate an apamin-sensitive medium AHP (mAHP) after each AP; and bursts of APs generate long-lasting slow AHPs (sAHPs) attributable to apamin-insensitive currents. Because all these currents are calcium dependent, we conducted voltage- and current-clamp whole-cell recordings while pharmacologically manipulating calcium channels of the plasma membrane and intracellular stores to determine what sources of calcium activate the currents underlying AP repolarization and the AHPs. The Cav2.2 (N-type) blocker omega conotoxin GVIA (1 microM) was the only blocker that significantly reduced the mAHP, and it induced a transition to rhythmic bursting in one-third of the cells tested. Cav1 (L-type) blockers (10 microM dihydropyridines) were the only ones that significantly reduced the sAHP. When applied to cells induced to burst with apamin, dihydropyridines reduced the sAHPs and abolished bursting. Depletion of intracellular stores with 10 mM caffeine also significantly reduced the sAHP current and reversibly regularized firing. Application of 1 microM omega conotoxin MVIIC (a Cav2.1/2.2 blocker) broadened APs but had a negligible effect on APs in cells in which BK channels were already blocked by submillimolar tetraethylammonium chloride, indicating that Cav2.1 (Q-type) channels provide the calcium to activate BK channels that repolarize the AP. Thus, calcium currents are selectively coupled to the calcium-dependent potassium currents underlying the AHPs, thereby creating mechanisms for control of the spontaneous firing patterns of these neurons. PMID- 16267232 TI - Neuronal L-type calcium channels open quickly and are inhibited slowly. AB - Neuronal L-type calcium channels are essential for regulating activity-dependent gene expression, but they are thought to open too slowly to contribute to action potential-dependent calcium entry. A complication of studying native L-type channels is that they represent a minor fraction of the whole-cell calcium current in most neurons. Dihydropyridine antagonists are therefore widely used to establish the contribution of L-type channels to various neuronal processes and to study their underlying biophysical properties. The effectiveness of these antagonists on L-type channels, however, varies with stimulus and channel subtype. Here, we study recombinant neuronal L-type calcium channels, CaV1.2 and CaV1.3. We show that these channels open with fast kinetics and carry substantial calcium entry in response to individual action potential waveforms, contrary to most studies of native L-type currents. Neuronal CaV1.3 L-type channels were as efficient as CaV2.2 N-type channels at supporting calcium entry during action potential-like stimuli. We conclude that the apparent slow activation of native L type currents and their lack of contribution to single action potentials reflect the state-dependent nature of the dihydropyridine antagonists used to study them, not the underlying properties of L-type channels. PMID- 16267233 TI - Development of a femtomolar-acting humanin derivative named colivelin by attaching activity-dependent neurotrophic factor to its N terminus: characterization of colivelin-mediated neuroprotection against Alzheimer's disease-relevant insults in vitro and in vivo. AB - Alzheimer's disease (AD) is the most common cause of dementia. Humanin (HN) is a short bioactive peptide abolishing neuronal cell death induced by various familial AD (FAD)-causative genes and amyloid-beta (Abeta) in vitro. It has been shown that HN suppresses memory impairment of mice induced by intracerebroventricular administration of Abeta. To potentiate the neuroprotective effect of HN, we synthesized a hybrid peptide named Colivelin composed of activity-dependent neurotrophic factor (ADNF) C-terminally fused to AGA-(C8R)HNG17, a potent HN derivative. Colivelin completely suppresses death induced by overexpressed FAD-causative genes and Abeta1-43 at a concentration of 100 fM, whereas AGA-(C8R)HNG17 does so at a concentration of 10 pM. Colivelin induced neuroprotection has been confirmed to occur via two neuroprotective pathways: one mediated by Ca2+/calmodulin-dependent protein kinase IV, triggered by ADNF, and one mediated by signal transducer and activator of transcription 3, triggered by HN. In vivo animal studies have further indicated that intracerebroventricular administration of Colivelin not only completely suppresses impairment in spatial working memory induced by repetitive intracerebroventricular injection of Abeta25-35 or Abeta1-42, but also it antagonizes neuronal loss in the CA1 region of hippocampus induced by hippocampal injection of Abeta1-42. In addition, intraperitoneally administered Colivelin suppresses memory impairment caused by a muscarinic acetylcholine receptor antagonist, 3-quinuclidinyl benzilate, indicating that a substantial portion of intraperitoneally administered Colivelin passes through the blood-brain barrier and suppresses functional memory deficit. Thus, Colivelin might serve as a novel drug candidate for treatment of AD. PMID- 16267234 TI - Apoptosis-inducing factor triggered by poly(ADP-ribose) polymerase and Bid mediates neuronal cell death after oxygen-glucose deprivation and focal cerebral ischemia. AB - Delayed neuronal cell death occurring hours after reperfusion is a hallmark of ischemic stroke and a primary target for neuroprotective strategies. In the present study, we investigated whether apoptosis-inducing factor (AIF), a caspase independent proapoptotic protein, is responsible for neuronal cell death after glutamate toxicity and oxygen-glucose deprivation (OGD) in vitro and after experimental stroke in vivo. AIF translocated to the nucleus in which it colocalized with DNA fragmentation and nuclear apoptotic morphology after exposure to glutamate or OGD in cultured neurons or after transient middle cerebral artery occlusion (MCAo) in mice. Small inhibitory RNA-mediated downregulation of AIF reduced glutamate- and OGD-induced neuronal apoptosis by 37 and 60%, respectively (p < 0.01). Moreover, Harlequin mutant mice, which express AIF at low levels (approximately 20% of wild-type mice), displayed smaller infarct volumes (-43%; p < 0.03) and showed dramatically reduced cell death in the ischemic penumbra after 45 min of MCAo compared with wild-type littermates. Inhibition of poly(ADP-ribose) polymerase and Bid reduced nuclear AIF translocation. These results provide the first evidence for a causal role of AIF in ischemic neuronal cell death. Therefore, caspase-independent cell death signaling may provide a promising novel target for therapeutic interventions in cerebrovascular diseases. PMID- 16267235 TI - Cholinergic deafferentation of the entorhinal cortex in rats impairs encoding of novel but not familiar stimuli in a delayed nonmatch-to-sample task. AB - Acetylcholine may regulate working memory for novel stimuli by activating intrinsic mechanisms for sustained spiking in entorhinal cortical neurons, which have been demonstrated in slice preparations of the entorhinal cortex. Computational modeling demonstrates that loss of the cholinergic activation of intrinsic mechanisms for sustained activity could selectively impair working memory for novel stimuli, whereas working memory for familiar stimuli could be maintained because of previously modified synapses. Blockade of muscarinic cholinergic receptors and selective cholinergic lesions has been shown to impair encoding in delayed matching tasks. However, previous studies have not compared explicitly the role of cholinergic modulation in working memory for novel versus familiar stimuli. Here, we show that lesions of the cholinergic innervation of the entorhinal cortex selectively impair delayed nonmatch to sample performance for novel odors, whereas delayed nonmatch to sample for familiar odors is spared. This indicates an important role for cholinergic innervation of the entorhinal cortex in working memory for novel stimuli. PMID- 16267237 TI - Function of atypical protein kinase C lambda in differentiating photoreceptors is required for proper lamination of mouse retina. AB - The photoreceptor is a highly polarized neuron and also has epithelial characteristics such as adherens junctions. To investigate the mechanisms of polarity formation of the photoreceptor cells, we conditionally knocked out atypical protein kinase Clambda (aPKClambda), which has been proposed to play a critical role in the establishment of epithelial and neuronal polarity, in differentiating photoreceptor cells using the Cre-loxP system. In aPKClambda conditional knock-out (CKO) mice, the photoreceptor cells displayed morphological defects and failed to form ribbon synapses. Intriguingly, lack of aPKClambda in differentiating photoreceptors led to severe laminar disorganization not only in the photoreceptor layer but also in the entire retina. Cell fate determination was not affected by total laminar disorganization. After Cre recombinase began to be expressed in the developing photoreceptors at embryonic day 12.5, both the immature photoreceptors and mitotic progenitors were dispersed throughout the CKO retina. We detected that adherens junction formation between the immature photoreceptors and the progenitors was lost in the CKO retina, whereas it was maintained between the progenitors themselves. These results indicate that the expression of aPKClambda in differentiating photoreceptors is required for total retinal lamination. Our data suggest that properly polarized photoreceptors anchor progenitors at the apical edge of the neural retina, which may be essential for building correct laminar organization of the retina. PMID- 16267236 TI - Rapid eye movement sleep is reduced in prolactin-deficient mice. AB - Prolactin (PRL) is implicated in the modulation of spontaneous rapid eye movement sleep (REMS). Previous models of hypoprolactinemic animals were characterized by changes in REMS, although associated deficits made it difficult to ascribe changes in REMS to reduced PRL. In the current studies, male PRL knock-out (KO) mice were used; these mice lack functional PRL but have no known additional deficits. Spontaneous REMS was reduced in the PRL KO mice compared with wild-type or heterozygous littermates. Infusion of PRL for 11-12 d into PRL KO mice restored their REMS to that occurring in wild-type or heterozygous controls. Six hours of sleep deprivation induced a non-REMS and a REMS rebound in both PRL KO mice and heterozygous littermates, although the REMS rebound in the KOs was substantially less. Vasoactive intestinal peptide (VIP) induced REMS responses in heterozygous mice but not in KO mice. Similarly, an ether stressor failed to enhance REMS in the PRL KOs but did in heterozygous littermates. Finally, hypothalamic mRNA levels for PRL, VIP, neural nitric oxide synthase (NOS), inducible NOS, and the interferon type I receptor were similar in KO and heterozygous mice. In contrast, tyrosine hydroxylase mRNA was lower in the PRL KO mice than in heterozygous controls and was restored to control values by infusion of PRL, suggesting a functioning short-loop negative feedback regulation in PRL KO mice. Data support the notion that PRL is involved in REMS regulation. PMID- 16267238 TI - Dynamically modulated spike correlation in monkey inferior temporal cortex depending on the feature configuration within a whole object. AB - The mechanism underlying the processing of spatially separated multiple local features to form a unique whole object is an important issue in visual object recognition. We tested whether, in behaving monkeys, the spike correlation between pairs of inferior temporal (IT) neurons dynamically changes depending on the spatial configuration of the local features within a whole object. We prepared more than 60,000 face-like objects (FOs) and their corresponding non face-like objects (NFOs) that consisted of random arrangements of the same set of local features as those in FOs. The spike correlation between a pair of neurons was quantified by the peak height of the shift predictor-subtracted cross correlogram. For both neurons of the pair, the local features in a whole object were determined so that they elicited as high a response as possible to enable a reliable cross-correlation analysis. We found that the FOs thus constructed elicited neuronal activities that were more strongly correlated than the corresponding NFOs. Firing rates of the same neurons did not show such a consistent bias depending on the feature configuration. Furthermore, receiver operating characteristic analysis revealed that this FO dominance of spike correlation was robust enough to discriminate between different feature configurations at the population level. Spike correlation of the cell pairs exhibited significant FO dominance within 300 ms after stimulus onset. The present results suggest that feature configuration within a unique whole object can be reflected in the rapid modulation of spike correlation among a population of neurons in the IT cortex. PMID- 16267239 TI - Cholinergic inhibition of neocortical pyramidal neurons. AB - Acetylcholine (ACh) is a central neurotransmitter critical for normal cognitive function. Here we show that transient muscarinic acetylcholine receptor activation directly inhibits neocortical layer 5 pyramidal neurons. Using whole cell and cell-attached recordings from neurons in slices of rat somatosensory cortex, we demonstrate that transient activation of M1-type muscarinic receptors induces calcium release from IP3-sensitive intracellular calcium stores and subsequent activation of an apamin-sensitive, SK-type calcium-activated potassium conductance. ACh-induced hyperpolarizing responses were blocked by atropine and pirenzepine but not by methoctramine or GABA receptor antagonists (picrotoxin, SR 95531 [2-(3-carboxypropyl)-3-amino-6-(4-methoxyphenyl)pyridazinium bromide], and CGP 55845 [(2S)-3-[[(15)-1-(3,4-dichlorophenyl)ethyl]amino-2 hydroxypropyl](phenylmethyl)phosphinic acid]). Responses were associated with a 31 +/- 5% increase in membrane conductance, had a reversal potential of -93 +/- 1 mV, and were eliminated after internal calcium chelation with BAPTA, blockade of IP3 receptors, or extracellular application of cadmium but not by sodium channel blockade with tetrodotoxin. Calcium-imaging experiments demonstrated that ACh induced hyperpolarizing, but not depolarizing, responses were correlated with large increases in intracellular calcium. Surprisingly, transient increases in muscarinic receptor activation were capable of generating hyperpolarizing responses even during periods of tonic muscarinic activation sufficient to depolarize neurons to action potential threshold. Furthermore, eserine, an acetylcholinesterase inhibitor similar to those used therapeutically in the treatment of Alzheimer's disease, disproportionately enhanced the excitatory actions of acetylcholine while reducing the ability of acetylcholine to generate inhibitory responses during repeated applications of ACh. These data demonstrate that acetylcholine can directly inhibit the output of neocortical pyramidal neurons. PMID- 16267240 TI - A small-molecule-inducible Nrf2-mediated antioxidant response provides effective prophylaxis against cerebral ischemia in vivo. AB - The transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2) coordinates expression of genes required for free radical scavenging, detoxification of xenobiotics, and maintenance of redox potential. Previously, activation of this pleiotropic response was neuroprotective in cell culture models that simulate components of stroke damage. However, the role of Nrf2 in limiting stroke damage in vivo remained unclear. We report that Nrf2 activation protects the brain from cerebral ischemia in vivo. Acute (1-3 d) intracerebroventricular or intraperitoneal pretreatment with tert butylhydroquinone (tBHQ), an Nrf2 activity inducer, reduced cortical damage and sensorimotor deficit at 24 h and even 1 month after ischemia-reperfusion in rats. Cortical glutathione levels robustly increased with tBHQ administration to rats and Nrf2-expressing mice, but not Nrf2(-/-) mice. Basal and inducible activities of antioxidant/detoxification enzymes in Nrf2(-/-) mice were reduced when compared with Nrf2(+/+) controls. Interestingly, larger infarcts were observed in Nrf2(-/-) mice at 7 d after stroke, but not at 24 h, suggesting that Nrf2 may play a role in shaping the penumbra well after the onset of ischemia. Neuronal death caused by a "penumbral" model of stroke, using intracortical endothelin-1 microinjection, was attenuated by tBHQ administration to Nrf2(+/+), but not to Nrf2(-/-) mice, confirming the Nrf2-specific action of tBHQ in vivo. We conclude that Nrf2 plays a role in modulating ischemic injury in vivo. Accordingly, Nrf2 activation by small molecule inducers may be a practical preventative treatment for stroke-prone patients. PMID- 16267241 TI - Cardiology patient page. Palpitations. PMID- 16267242 TI - Images in cardiovascular medicine. The catch: a graphic demonstration of the value of embolic protection devices. PMID- 16267243 TI - Images in cardiovascular medicine. Novel magnetic resonance-compatible coronary stent: the absorbable magnesium-alloy stent. PMID- 16267244 TI - Images in cardiovascular medicine. Dehiscence of mitral annuloplasty ring. PMID- 16267245 TI - Letter regarding article by Valgimigli et al, "Short- and long-term clinical outcome after drug-eluting stent implantation for the percutaneous treatment of left main coronary artery disease". PMID- 16267246 TI - Guidelines, lighthouses, and a toe in the water. PMID- 16267247 TI - Evolving therapeutic strategies for dystrophinopathies: potential for conflict between cardiac and skeletal needs. PMID- 16267248 TI - Prevention of restenosis by systemic drug therapy: back to the future? PMID- 16267249 TI - Prevention of fatal arrhythmias in high-risk subjects by fish oil n-3 fatty acid intake. AB - BACKGROUND: The long-chain n-3 fatty acids in fish have been demonstrated to have antiarrhythmic properties in experimental models and to prevent sudden cardiac death in a randomized trial of post-myocardial infarction patients. Therefore, we hypothesized that these n-3 fatty acids might prevent potentially fatal ventricular arrhythmias in high-risk patients. METHODS AND RESULTS: Four hundred two patients with implanted cardioverter/defibrillators (ICDs) were randomly assigned to double-blind treatment with either a fish oil or an olive oil daily supplement for 12 months. The primary end point, time to first ICD event for ventricular tachycardia or fibrillation (VT or VF) confirmed by stored electrograms or death from any cause, was analyzed by intention to treat. Secondary analyses were performed for "probable" ventricular arrhythmias, "on treatment" analyses for all subjects who had taken any of their oil supplements, and "on-treatment" analyses only of those subjects who were on treatment for at least 11 months. Compliance with double-blind treatment was similar in the 2 groups; however, the noncompliance rate was high (35% of all enrollees). In the primary analysis, assignment to treatment with the fish oil supplement showed a trend toward a prolonged time to the first ICD event (VT or VF) or of death from any cause (risk reduction of 28%; P=0.057). When therapies for probable episodes of VT or VF were included, the risk reduction became significant at 31%; P=0.033. For those who stayed on protocol for at least 11 months, the antiarrhythmic benefit of fish oil was improved for those with confirmed events (risk reduction of 38%; P=0.034). CONCLUSIONS: Although significance was not achieved for the primary end point, this study provides evidence that for individuals at high risk of fatal ventricular arrhythmias, regular daily ingestion of fish oil fatty acids may significantly reduce potentially fatal ventricular arrhythmias. PMID- 16267250 TI - Right ventricular fibrosis and conduction delay in a patient with clinical signs of Brugada syndrome: a combined electrophysiological, genetic, histopathologic, and computational study. AB - BACKGROUND: The mechanism of ECG changes and arrhythmogenesis in Brugada syndrome (BS) patients is unknown. METHODS AND RESULTS: A BS patient without clinically detected cardiac structural abnormalities underwent cardiac transplantation for intolerable numbers of implantable cardioverter/defibrillator discharges. The patient's explanted heart was studied electrophysiologically and histopathologically. Whole-cell currents were measured in HEK293 cells expressing wild-type or mutated sodium channels from the patient. The right ventricular outflow tract (RVOT) endocardium showed activation slowing and was the origin of ventricular fibrillation without a transmural repolarization gradient. Conduction restitution was abnormal in the RVOT but normal in the left ventricle. Right ventricular hypertrophy and fibrosis with epicardial fatty infiltration were present. HEK293 cells expressing a G1935S mutation in the gene encoding the cardiac sodium channel exhibited enhanced slow inactivation compared with wild type channels. Computer simulations demonstrated that conduction slowing in the RVOT might have been the cause of the ECG changes. CONCLUSIONS: In this patient with BS, conduction slowing based on interstitial fibrosis, but not transmural repolarization differences, caused the ECG signs and was the origin of ventricular fibrillation. PMID- 16267251 TI - Pulmonary neovascularity: a distinctive radiographic finding in Eisenmenger syndrome. AB - BACKGROUND: We sought to characterize the distinctive pulmonary vascular abnormalities seen on chest radiographs and computed tomography (CT) scans in Eisenmenger syndrome. METHODS AND RESULTS: Thoracic CT scans, chest radiographs, and clinical data were reviewed for 24 Eisenmenger syndrome patients subdivided into those with interatrial (pretricuspid) versus interventricular or great arterial (posttricuspid) communications and in 14 acyanotic patients with pulmonary arterial hypertension (PAH) and no congenital heart disease. CT scans were scored blindly by 2 thoracic radiologists for the presence and severity of small, tortuous, intrapulmonary vessels, termed "neovascularity," lobular ground glass opacification, and systemic perihilar and intercostal vessels. Histopathologic lung sections from 5 patients with Eisenmenger syndrome and from 3 patients with acyanotic PAH were reviewed. Associations between clinical and imaging features were tested by ANOVA and chi2 tests. Kendall's rank-order coefficient and the Kruskal-Wallis test were used to test for significant differences in imaging features between Eisenmenger syndrome and acyanotic PAH. Neovascularity on chest radiographs was more common in Eisenmenger syndrome than acyanotic PAH, but differences were not significant. On CT, neovascularity, lobular ground-glass opacification, and hilar and intercostal systemic collaterals were more prevalent in Eisenmenger syndrome, with severity greater in posttricuspid communications. Three previously undescribed vascular lesions were identified histologically in Eisenmenger syndrome: malformed, dilated, muscular arteries within alveolar septa; congested capillaries within alveolar walls; and congested capillaries within the walls of medium-size, muscular pulmonary arteries. These lesions may correspond to the distinctive vascular abnormalities observed on chest radiographs and CT scans. CONCLUSIONS: Distinctive vascular lesions on chest radiographs and CT scans in Eisenmenger syndrome appear to be correlated histologically with collateral vessels that develop more extensively with posttricuspid communications. PMID- 16267252 TI - Relationship between procedure indications and outcomes of percutaneous coronary interventions by American College of Cardiology/American Heart Association Task Force Guidelines. AB - BACKGROUND: An American College of Cardiology/American Heart Association (ACC/AHA) Task Force periodically revises and publishes guidelines with evidence based recommendations for appropriate use of percutaneous coronary intervention (PCI). Some studies have suggested that closer adherence to guidelines can reduce variations in care, can improve quality, and may ultimately result in better outcomes, but this finding is incompletely understood. Guidelines themselves must change to be responsive to continuously evolving clinical practice. Our goal here was to investigate whether any relationship existed between the most recent ACC/AHA recommended indications for PCI and short term in-hospital outcomes. METHODS AND RESULTS: We analyzed the ACC National Cardiovascular Data Registry for the period of January 1, 2001, through March 31, 2004. We excluded PCI procedures performed for acute myocardial infarction (ST-segment elevation myocardial infarction); all others were grouped by their indications according to the standard ACC/AHA scheme: Class I, evidence and/or agreement that PCI is useful and effective; Class IIa, conflicting evidence and/or divergent opinions, weight is in favor; Class IIb, usefulness/efficacy is less well established; and Class III, evidence and/or agreement that PCI is not useful or effective and may be harmful. Clinical success was defined as angiographic success (<20% residual stenosis) at all lesions attempted without the adverse events of myocardial infarction, same-admission bypass surgery, or death. There were 412 617 PCI procedures included in the analysis. Frequency of indications was as follows: Class I, 64%; Class IIa, 21%; Class IIb, 7%; and Class III, 8%. Clinical success declined across the indications classes (92.8%, 91.7%, 89%, and 85.5%, respectively; P<0.001), whereas adverse events increased. CONCLUSIONS: In this large survey of contemporary PCI practice, most procedures were performed for Class I indications. A significant relationship between evidence-based indications recommended by the ACC/AHA Task Force and in-hospital outcomes was noted. PMID- 16267253 TI - Gene mutations in apical hypertrophic cardiomyopathy. AB - BACKGROUND: Nonobstructive hypertrophy localized to the cardiac apex is an uncommon morphological variant of hypertrophic cardiomyopathy (HCM) that often is further distinguished by distinct giant negative T waves and a benign clinical course. The genetic relationship between HCM with typical hypertrophic morphology versus isolated apical hypertrophy is incompletely understood. METHODS AND RESULTS: Genetic cause was investigated in 15 probands with apical hypertrophy by DNA sequence analyses of 9 sarcomere protein genes and 3 other genes (GLA, PRKAG2, and LAMP2) implicated in idiopathic cardiac hypertrophy. Six sarcomere gene mutations were found in 7 samples; no samples contained mutations in GLA, PRKAG2, or LAMP2. Clinical evaluations demonstrated familial apical HCM in 4 probands, and in 3 probands disease-causing mutations were identified. Two families shared a cardiac actin Glu101Lys missense mutation; all members of both families with clinical manifestations of HCM (n=16) had apical hypertrophy. An essential light chain missense mutation Met149Val caused apical or midventricular segment HCM in another proband and 5 family members, but 6 other affected relatives had typical HCM morphologies. No other sarcomere gene mutations identified in the remaining probands caused apical HCM in other family members. CONCLUSIONS: Sarcomere protein gene mutations that cause apical hypertrophy rather than more common HCM morphologies reflect interactions among genetic etiology, background modifier genes, and/or hemodynamic factors. Only a limited number of sarcomere gene defects (eg, cardiac actin Glu101Lys) consistently produce apical HCM. PMID- 16267254 TI - Myeloperoxidase-generated oxidants modulate left ventricular remodeling but not infarct size after myocardial infarction. AB - BACKGROUND: Inflammation after myocardial infarction (MI) heralds worse left ventricular (LV) function and clinical outcomes. However, whether inflammation affects LV function by extending myonecrosis and/or altering LV remodeling remains unknown. We hypothesized that cytotoxic aldehydes generated during oxidative stress may adversely affect remodeling and infarct size. One theoretical source of reactive aldehydes is oxidation of common alpha-amino acids by myeloperoxidase (MPO) released by leukocytes. However, a role for MPO in formation of aldehydes in vivo and the functional consequences of MPO-generated oxidants in ischemia/reperfusion models of MI have not been established. METHODS AND RESULTS: In studies with cell types found in vascular tissue, MPO-oxidation products of glycine (formaldehyde) and threonine (acrolein) were the most cytotoxic. Mass spectrometry studies of myocardial tissue from murine models of acute MI (both chronic left anterior descending coronary artery ligation and ischemia/reperfusion injury) confirmed that MPO serves as a major enzymatic source in the generation of these cytotoxic aldehydes. Interestingly, although MPO-null mice experienced 35.1% (P<0.001) less LV dilation and a 52.2% (P<0.0001) improvement in LV function compared with wild-type mice 24 days after ischemia/reperfusion injury, no difference in infarct size between wild-type and MPO-null mice was noted. CONCLUSIONS: The present data separate inflammatory effects on infarct size and LV remodeling and demonstrate that MPO-generated oxidants do not significantly affect tissue necrosis after MI but rather have a profound adverse effect on LV remodeling and function. PMID- 16267255 TI - Magnetic resonance assessment of the substrate for inducible ventricular tachycardia in nonischemic cardiomyopathy. AB - BACKGROUND: Patients with left ventricular dysfunction have an elevated risk of sudden cardiac death. However, the substrate for ventricular arrhythmia in patients with nonischemic cardiomyopathy remains poorly understood. We hypothesized that the distribution of scar identified by MRI is predictive of inducible ventricular tachycardia. METHODS AND RESULTS: Short-axis cine steady state free-precession and postcontrast inversion-recovery gradient-echo MRI sequences were obtained before electrophysiological study in 26 patients with nonischemic cardiomyopathy. Left ventricular ejection fraction was measured from end-diastolic and end-systolic cine images. The transmural extent of scar as a percentage of wall thickness (percent scar transmurality) in each of 12 radial sectors per slice was calculated in all myocardial slices. The percentages of sectors with 1% to 25%, 26% to 50%, 51% to 75%, and 76% to 100% scar transmurality were determined for each patient. Predominance of scar distribution involving 26% to 75% of wall thickness was significantly predictive of inducible ventricular tachycardia and remained independently predictive in the multivariable model after adjustment for left ventricular ejection fraction (odds ratio, 9.125; P=0.020). CONCLUSIONS: MR assessment of scar distribution can identify the substrate for inducible ventricular tachycardia and may identify high-risk patients with nonischemic cardiomyopathy currently missed by ejection fraction criteria. PMID- 16267256 TI - Robustness of late lumen loss in discriminating drug-eluting stents across variable observational and randomized trials. AB - BACKGROUND: Binary angiographic and clinical restenosis rates can vary widely between clinical studies, even for the same stent, influenced heavily by case-mix covariates that differ among observational and randomized trials intended to assess a given stent system. We hypothesized that mean in-stent late loss might be a more stable estimator of restenosis propensity across such studies. METHODS AND RESULTS: In 46 trials of drug-eluting and bare-metal stenting, increasing mean late loss was associated with higher target lesion revascularization (TLR) rates (P<0.001). When the class of bare-metal stents was compared with the class of effective drug-eluting stents, late loss was more discriminating than TLR as measured by the high intraclass correlation coefficient (rho) (late loss, rho=0.71 versus TLR, rho=0.22; 95% CI of difference=0.33, 0.65). When the individual drug-eluting stents and bare-metal stents were compared, late loss was a better discriminator than TLR (0.68 versus 0.19; 95% CI of difference=0.24, 0.60). Greater adjustments of study covariates are needed to stabilize assessments of TLR compared with late loss because of greater influence of reference vessel diameter on TLR than on in-stent late loss. Optimization of late loss with the use of a novel method of standardization according to diabetes prevalence and mean lesion length resulted in minor adjustments in late loss (<0.08 mm for 90% of reported trials) and an ordered array of mean late loss values for the stent systems studied. CONCLUSIONS: Late loss is more reliable than restenosis rates for discriminating restenosis propensity between new drug eluting stent platforms across studies and might be the optimum end point for evaluating drug-eluting stents in early, nonrandomized studies. PMID- 16267257 TI - Id1 gene transfer confers angiogenic property on fully differentiated endothelial cells and contributes to therapeutic angiogenesis. AB - BACKGROUND: Transplantation of endothelial progenitor cells has been proposed as a potential strategy for therapeutic revascularization. However, the limited endogenous cell pool and the related technical difficulties constitute clinically important disadvantages to autologous transplantation. In this study we investigated whether fully differentiated endothelial cells (ECs) modified with gene transfer of Id1, a helix-loop-helix transcription factor involved in angiogenesis, have the potential to contribute to therapeutic angiogenesis. METHODS AND RESULTS: The Id1 gene was transferred into human umbilical vein ECs (HUVECs) via a Sendai virus vector. Id1 stimulated migration, proliferation, and capillary-like tube/cord formation of HUVECs. In addition, Id1 reduced serum deprivation-induced HUVEC apoptosis, as shown by FACS analysis with annexin V and TUNEL staining. Transplantation of Id1-overexpressing HUVECs accelerated recovery of blood flow as evaluated by laser-Doppler perfusion imaging, increased capillary density, and improved the rate of limb salvage compared with the transplantation of control HUVECs. Histochemical analysis revealed that the regenerated vascular networks of limbs transplanted with Id1-overexpressing HUVECs contained numerous HUVECs, some of which were in a proliferative state. Untransfected HUVECs were also incorporated with Id1-transfected HUVECs, suggesting the noncell autonomous effect of Id1. Finally, angiopoietin-1 was upregulated in Id1-overexpressing HUVECs and functionally contributed to the in vitro angiogenic effect of Id1. CONCLUSIONS: Id1 gene transfer conferred HUVECs with an angiogenic property, contributing to neovascularization after transplantation into ischemic lesions. Transplantation of Id1-overexpressing mature ECs may serve as a novel and useful strategy for therapeutic angiogenesis. PMID- 16267258 TI - Cardiac troponin elevation, cardiovascular morbidity, and outcome after subarachnoid hemorrhage. AB - BACKGROUND: Cardiac troponin I (cTI) release occurs frequently after subarachnoid hemorrhage (SAH) and has been associated with a neurogenic form of myocardial injury. The prognostic significance and clinical impact of these elevations remain poorly defined. METHODS AND RESULTS: We studied 253 SAH patients who underwent serial cTI measurements for clinical or ECG signs of potential cardiac injury. These patients were drawn from an inception cohort of 441 subjects enrolled in the Columbia University SAH Outcomes Project between November 1998 and August 2002. Peak cTI levels were divided into quartiles or classified as undetectable. Adverse in-hospital events were prospectively recorded, and outcome at 3 months was assessed with the modified Rankin Scale. Admission predictors of cTI elevation included poor clinical grade, intraventricular hemorrhage, loss of consciousness at ictus, global cerebral edema, and a composite score of physiological derangement (all P< or =0.01). Peak cTI level was associated with an increased risk of echocardiographic left ventricular dysfunction (odds ratio [OR], 1.3 per quintile; 95% CI, 1.0 to 1.7; P=0.03), pulmonary edema (OR, 2.1 per quintile; 95% CI, 1.6 to 2.7; P<0.001), hypotension requiring pressors (OR, 1.9 per quintile; 95% CI, 1.5 to 2.3; P<0.001), and delayed cerebral ischemia from vasospasm (OR, 1.3 per quintile; 95% CI, 1.07 to 1.7; P=0.01). Peak cTI levels were predictive of death or severe disability at discharge after controlling for age, clinical grade, and aneurysm size (adjusted OR, 1.4 per quintile; 95% CI, 1.1 to 1.9; P=0.02), but this association was no longer significant at 3 months. CONCLUSIONS: cTI elevation after SAH is associated with an increased risk of cardiopulmonary complications, delayed cerebral ischemia, and death or poor functional outcome at discharge. PMID- 16267259 TI - Involvement of thromboxane receptor in the proatherogenic effect of isoprostane F2alpha-III: evidence from apolipoprotein E- and LDL receptor-deficient mice. AB - BACKGROUND: Atherosclerosis is a chronic inflammatory disease of the arterial wall, where it associates with oxidative stress and formation of oxidized lipids. The lipid oxidation product isoprostane iPF2alpha-III, also known as 8 isoPGF2alpha and 15-F2t-IsoP, is elevated in patients with cardiovascular disease and present in atherosclerotic lesions. Several proatherogenic biological effects have been attributed to this isoprostane, suggesting that it could be an active factor in the pathogenesis of the disease. METHODS AND RESULTS: In this study we show that iPF2alpha-III directly promotes atherogenesis in 2 different mouse models (ie, apolipoprotein E [apoE]- and LDL receptor-deficient mice) by activating the thromboxane receptor (TP). This effect is mediated by potent proinflammatory vascular reactions but is independent of thromboxane A2 levels, changes in blood pressure, or lipid profile. Pharmacological antagonism of TP suppresses the vascular proatherogenic effects of iPF2alpha-III. Endothelial cells genetically lacking TP show reduced inflammatory responses when stimulated with this product of lipid oxidation but not other oxidized lipids. CONCLUSIONS: Our results demonstrate that in atherosclerosis iPF2alpha-III is not only a biomarker of oxidative stress but also an active mediator of its vascular phenotype. We conclude that in a clinical setting in which both thromboxane A2 and iPF2alpha-III are elevated, suppression of the first alone would not provide the most benefit for patients because the coincidental presence of the isoprostane will still have a proatherogenic effect. PMID- 16267260 TI - Sevelamer prevents uremia-enhanced atherosclerosis progression in apolipoprotein E-deficient mice. AB - BACKGROUND: The novel phosphate binder sevelamer has been shown to prevent the progression of aortic and coronary calcification in uremic patients. Whether it also decreases the progression of atheromatous plaques is unknown. The aim of our study was to examine the effect of sevelamer administration on the development of atherosclerosis and aortic calcification in the uremic apolipoprotein E-deficient mouse as an established model of accelerated atherosclerosis. METHODS AND RESULTS: Female mice were randomly assigned to 4 groups: 2 groups of nonuremic mice (sevelamer versus control) and 2 groups of uremic mice (sevelamer versus control). Sevelamer was given at 3% with chow. The increases in serum phosphorus concentration and calcium-phosphorus product observed in uremic control mice were prevented by sevelamer. Serum total cholesterol was increased in the 2 uremic mouse groups and remained unchanged in response to sevelamer. After 8 weeks of sevelamer treatment, uremic mice exhibited a significantly lower degree of atherosclerosis (P<0.001) and vascular calcification than uremic control mice. Of interest, sevelamer exerted an effect on both intima and media calcification (P=0.005) in uremic mice. Among possible mechanisms involved, we found no evidence for the modulation by sevelamer of inflammation or selected uremic toxins. In contrast, nitrotyrosine staining as a measure of oxidative damage was significantly decreased in response to sevelamer treatment in control and uremic mice (P<0.005). CONCLUSIONS: Sevelamer delays not only vascular calcification but also atherosclerotic lesion progression in uremic apolipoprotein E-deficient mice. It opens the possibility of a cholesterol-independent action of sevelamer on atheroma formation via effects on mineral metabolism, oxidative stress, or both. PMID- 16267261 TI - Humanitarian Use Devices/Humanitarian Device Exemptions in cardiovascular medicine. AB - The Second Dartmouth Device Development Symposium held in October 2004 brought together leaders from the medical device community, including clinical investigators, senior representatives from the US Food and Drug Administration, large and small device manufacturers, and representatives from the financial community to examine difficult issues confronting device development. The role of the Humanitarian Use Device/Humanitarian Device Exemption (HUD/HDE) pathway in the development of new cardiovascular devices was discussed in this forum. The HUD/HDE pathway was created by Congress to facilitate the availability of medical devices for "orphan" indications, ie, those affecting <4000 individuals within the United States each year. The HUD/HDE pathway streamlines the approval process and permits less well-characterized devices to enter the market. HDE approval focuses primarily on issues of safety and scientific soundness and does not require demonstration of efficacy. In the 7 years since the first device was approved in 1997, a total of 35 HDEs have been granted (23 devices, 6 diagnostic tests). As the costs to gain regulatory approval for commonly used devices increase, companies often seek alternative ways to gain market access, including the HUD/HDE pathway. For a given device, there may be multiple legitimate and distinct indications, including indications that meet the HUD criteria. Companies must choose how and when to pursue each of these indications. The consensus of symposium participants was for the HUD/HDE pathway to be reserved for true orphan indications and not be viewed strategically as part of the clinical development plan to access a large market. PMID- 16267262 TI - Hematology, ASH, and the anemia of the aged. PMID- 16267263 TI - Gamma-glutamyl carboxylase (GGCX) microsatellite and warfarin dosing. PMID- 16267264 TI - Role of VAD in the initial treatment of multiple myeloma. PMID- 16267265 TI - CAR-1, a protein that localizes with the mRNA decapping component DCAP-1, is required for cytokinesis and ER organization in Caenorhabditis elegans embryos. AB - The division of one cell into two requires the coordination of multiple components. We describe a gene, car-1, whose product may provide a link between disparate cellular processes. Inhibition of car-1 expression in Caenorhabditis elegans embryos causes late cytokinesis failures: cleavage furrows ingress but subsequently regress and the spindle midzone fails to form, even though midzone components are present. The localized accumulation of membrane that normally develops at the apex of the cleavage furrow during the final phase of cytokinesis does not occur and organization of the endoplasmic reticulum is aberrant, indicative of a disruption in membrane trafficking. The car-1 gene has homologues in a number of species, including proteins that associate with RNA binding proteins. CAR-1 localizes to P-granules (germ-line specific ribonucleoprotein particles) and discrete, developmentally regulated cytoplasmic foci. These foci also contain DCAP-1, a protein involved in decapping mRNAs. Thus, CAR-1, a protein likely to be associated with RNA metabolism, plays an essential role in the late stage of cytokinesis, suggesting a novel link between RNA, membrane trafficking and cytokinesis in the C. elegans embryo. PMID- 16267266 TI - Molecular interaction maps of bioregulatory networks: a general rubric for systems biology. AB - A standard for bioregulatory network diagrams is urgently needed in the same way that circuit diagrams are needed in electronics. Several graphical notations have been proposed, but none has become standard. We have prepared many detailed bioregulatory network diagrams using the molecular interaction map (MIM) notation, and we now feel confident that it is suitable as a standard. Here, we describe the MIM notation formally and discuss its merits relative to alternative proposals. We show by simple examples how to denote all of the molecular interactions commonly found in bioregulatory networks. There are two forms of MIM diagrams. "Heuristic" MIMs present the repertoire of interactions possible for molecules that are colocalized in time and place. "Explicit" MIMs define particular models (derived from heuristic MIMs) for computer simulation. We show also how pathways or processes can be highlighted on a canonical heuristic MIM. Drawing a MIM diagram, adhering to the rules of notation, imposes a logical discipline that sharpens one's understanding of the structure and function of a network. PMID- 16267267 TI - Different Plk1 functions show distinct dependencies on Polo-Box domain-mediated targeting. AB - Polo-like kinase 1 (Plk1) has multiple important functions during M-phase progression. In addition to a catalytic domain, Plk1 possesses a phosphopeptide binding motif, the polo-box domain (PBD), which is required for proper localization. Here, we have explored the importance of correct Plk1 subcellular targeting for its mitotic functions. We either displaced endogenous Plk1 through overexpression of the PBD or introduced the catalytic domain of Plk1, lacking the PBD, into Plk1-depleted cells. Both treatments resulted in remarkably similar phenotypes, which were distinct from the Plk1 depletion phenotype. Cells depleted of Plk1 mostly arrested with monoastral spindles, because of inhibition of centrosome maturation and separation. In contrast, these functions were not impaired in cells with mislocalized Plk1. Instead, these latter cells showed a checkpoint-dependent mitotic arrest characterized by impaired chromosome congression. Thus, whereas chromosome congression requires localized Plk1 activity, other investigated Plk1 functions are less dependent on correct PBD mediated targeting. This opens the possibility that PBD-directed drugs might be developed to selectively interfere with a subset of Plk1 functions. PMID- 16267269 TI - Phosphoinositide 3-kinase activity controls the chemoattractant-mediated activation and adaptation of adenylyl cyclase. AB - The binding of chemoattractants to cognate G protein-coupled receptors activates a variety of signaling cascades that provide spatial and temporal cues required for chemotaxis. When subjected to uniform stimulation, these responses are transient, showing an initial peak of activation followed by a period of adaptation, in which activity subsides even in the presence of stimulus. A tightly regulated balance between receptor-mediated stimulatory and inhibitory pathways controls the kinetics of activation and subsequent adaptation. In Dictyostelium, the adenylyl cyclase expressed during aggregation (ACA), which synthesizes the chemoattractant cAMP, is essential to relay the signal to neighboring cells. Here, we report that cells lacking phosphoinositide 3-kinase (PI3K) activity are deficient in signal relay. In LY294002-treated cells, this defect is because of a loss of ACA activation. In contrast, in cells lacking PI3K1 and PI3K2, the signal relay defect is because of a loss of ACA adaptation. We propose that the residual low level of 3-phosphoinositides in pi3k(1-/2-) cells is sufficient to generate the initial peak of ACA activity, yet is insufficient to sustain the inhibitory phase required for its adaptation. Thus, PI3K activity is poised to regulate both ACA activation and adaptation, thereby providing a link to ensure the proper balance of counteracting signals required to maintain optimal chemoresponsiveness. PMID- 16267268 TI - Slx4 regulates DNA damage checkpoint-dependent phosphorylation of the BRCT domain protein Rtt107/Esc4. AB - RTT107 (ESC4, YHR154W) encodes a BRCA1 C-terminal-domain protein that is important for recovery from DNA damage during S phase. Rtt107 is a substrate of the checkpoint protein kinase Mec1, although the mechanism by which Rtt107 is targeted by Mec1 after checkpoint activation is currently unclear. Slx4, a component of the Slx1-Slx4 structure-specific nuclease, formed a complex with Rtt107. Deletion of SLX4 conferred many of the same DNA-repair defects observed in rtt107delta, including DNA damage sensitivity, prolonged DNA damage checkpoint activation, and increased spontaneous DNA damage. These phenotypes were not shared by the Slx4 binding partner Slx1, suggesting that the functions of the Slx4 and Slx1 proteins in the DNA damage response were not identical. Of particular interest, Slx4, but not Slx1, was required for phosphorylation of Rtt107 by Mec1 in vivo, indicating that Slx4 was a mediator of DNA damage dependent phosphorylation of the checkpoint effector Rtt107. We propose that Slx4 has roles in the DNA damage response that are distinct from the function of Slx1 Slx4 in maintaining rDNA structure and that Slx4-dependent phosphorylation of Rtt107 by Mec1 is critical for replication restart after alkylation damage. PMID- 16267271 TI - An uncleavable uPAR mutant allows dissection of signaling pathways in uPA dependent cell migration. AB - Urokinase-type plasminogen activator (uPA) binding to uPAR induces migration, adhesion, and proliferation through multiple interactions with G proteins-coupled receptor FPRL1, integrins, or the epidermal growth factor (EGF) receptor (EGFR). At least two forms of uPAR are present on the cell surface: full-length and cleaved uPAR, each specifically interacting with one or more transmembrane proteins. The connection between these interactions and the effects on the signaling pathways activation is not clear. We have exploited an uPAR mutant (hcr, human cleavage resistant) to dissect the pathways involved in uPA-induced cell migration. This mutant is not cleaved by proteases, is glycosylphosphatidylinositol anchored, and binds uPA with a normal K(d). Both wild-type (wt) and hcr-uPAR are able to mediate uPA-induced migration, are constitutively associated with the EGFR, and associate with alpha3beta1 integrin upon uPA binding. However, they engage different pathways in response to uPA. wt uPAR requires both integrins and FPRL1 to mediate uPA-induced migration, and association of wt-uPAR to alpha3beta1 results in uPAR cleavage and extracellular signal-regulated kinase (ERK) activation. On the contrary, hcr-uPAR does not activate ERK and does not engage FPRL1 or any other G protein-coupled receptor, but it activates an alternative pathway initiated by the formation of a triple complex (uPAR-alpha3beta1-EGFR) and resulting in the autotyrosine phosphorylation of EGFR. PMID- 16267270 TI - Binding of Src to Na+/K+-ATPase forms a functional signaling complex. AB - We have shown that ouabain activates Src, resulting in subsequent tyrosine phosphorylation of multiple effectors. Here, we tested if the Na+/K+-ATPase and Src can form a functional signaling complex. In LLC-PK1 cells the Na+/K+-ATPase and Src colocalized in the plasma membrane. Fluorescence resonance energy transfer analysis indicated that both proteins were in close proximity, suggesting a direct interaction. GST pulldown assay showed a direct, ouabain regulated, and multifocal interaction between the 1 subunit of Na+/K+-ATPase and Src. Although the interaction between the Src kinase domain and the third cytosolic domain (CD3) of 1 is regulated by ouabain, the Src SH3SH2 domain binds to the second cytosolic domain constitutively. Functionally, binding of Src to either the Na+/K+-ATPase or GST-CD3 inhibited Src activity. Addition of ouabain, but not vanadate, to the purified Na+/K+-ATPase/Src complex freed the kinase domain and restored the Src activity. Consistently, exposure of intact cells to ouabain apparently increased the distance between the Na+/K+-ATPase and Src. Concomitantly, it also stimulated tyrosine phosphorylation of the proteins that are associated with the Na+/K+-ATPase. These new findings illustrate a novel molecular mechanism of signal transduction involving the interaction of a P-type ATPase and a nonreceptor tyrosine kinase. PMID- 16267272 TI - The flagellar motility of Chlamydomonas pf25 mutant lacking an AKAP-binding protein is overtly sensitive to medium conditions. AB - Radial spokes are a conserved axonemal structural complex postulated to regulate the motility of 9 + 2 cilia and flagella via a network of phosphoenzymes and regulatory proteins. Consistently, a Chlamydomonas radial spoke protein, RSP3, has been identified by RII overlays as an A-kinase anchoring protein (AKAP) that localizes the cAMP-dependent protein kinase (PKA) holoenzyme by binding to the RIIa domain of PKA RII subunit. However, the highly conserved docking domain of PKA is also found in the N termini of several AKAP-binding proteins unrelated to PKA as well as a 24-kDa novel spoke protein, RSP11. Here, we report that RSP11 binds to RSP3 directly in vitro and colocalizes with RSP3 toward the spoke base near outer doublets and dynein motors in axonemes. Importantly, RSP11 mutant pf25 displays a spectrum of motility, from paralysis with flaccid or twitching flagella as other spoke mutants to wildtype-like swimming. The wide range of motility changes reversibly depending on the condition of liquid media without replacing defective proteins. We postulate that radial spokes use the RIIa/AKAP module to regulate ciliary and flagellar beating; absence of the spoke RIIa protein exposes a medium-sensitive regulatory mechanism that is not obvious in wild-type Chlamydomonas. PMID- 16267273 TI - Conserved prefusion protein assembly in regulated exocytosis. AB - The regulated release of hormones and neurotransmitters is a fundamental process throughout the animal kingdom. The short time scale for the calcium triggering of vesicle fusion in regulated secretion suggests that the calcium sensor synaptotagmin and the soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) membrane fusion machinery are well ordered before the calcium signal. To gain insight into the organization of the prefusion protein assembly in regulated exocytosis, we undertook a structural/functional study of the vesicular synaptotagmin1 and the plasma membrane SNARE proteins, which copurify from the brain in the absence of calcium. Based on an evolutionary analysis, mutagenesis screens, and a computational protein docking approach, we now provide the first testable description of the supramolecular prefusion assembly. Perturbing the determined synaptotagmin/SNARE-interacting interface in several models of regulated exocytosis altered the secretion of hormones and neurotransmitters. These mutations also disrupted the constitutive synaptotagmin/SNARE link in full agreement with our model. We conclude that the interaction of synaptotagmin with preassembled plasma membrane SNARE proteins, before the action of calcium, can provide a precisely organized "tethering" scaffold that underlies regulated secretion throughout evolution. PMID- 16267275 TI - Rescue of vasopressin V2 receptor mutants by chemical chaperones: specificity and mechanism. AB - Because missense mutations in genetic diseases of membrane proteins often result in endoplasmic reticulum (ER) retention of functional proteins, drug-induced rescue of their cell surface expression and understanding the underlying mechanism are of clinical value. To study this, we tested chemical chaperones and sarco(endo)plasmic reticulum Ca2+ ATPase pump inhibitors on Madin-Darby canine kidney cells expressing nine ER-retained vasopressin type-2 receptor (V2R) mutants involved in nephrogenic diabetes insipidus. Of these nine, only V2R-V206D showed improved maturation and plasma membrane rescue with glycerol, dimethyl sulfoxide (DMSO), thapsigargin/curcumin, and ionomycin but not with other osmolytes or growth at 27 degrees C. This revealed that rescue is mutant specific and that this mutant is prone to rescue by multiple compounds. Rescue did not involve changed expression of molecular chaperones calnexin, heat-shock protein (HSP) 70, or HSP90. V2R antagonist SR121463B treatment revealed that V2R-V206D and V2R-S167T were rescued and matured to a greater extent, suggesting that the rescuing activity of a pharmacological versus chemical chaperone is broader and stronger. Calcium measurements showed that rescue of V2R-V206D by thapsigargin, curcumin, and ionomycin was because of increased cytosolic calcium level, rather than decreased endoplasmic reticulum calcium level. The molecular mechanism underlying rescue by DMSO, glycerol, and SR121463B is different, because with these compounds intracellular calcium levels were unaffected. PMID- 16267274 TI - A genomewide screen for petite-negative yeast strains yields a new subunit of the i-AAA protease complex. AB - Unlike many other organisms, the yeast Saccharomyces cerevisiae can tolerate the loss of mitochondrial DNA (mtDNA). Although a few proteins have been identified that are required for yeast cell viability without mtDNA, the mechanism of mtDNA independent growth is not completely understood. To probe the relationship between the mitochondrial genome and cell viability, we conducted a microarray based, genomewide screen for mitochondrial DNA-dependent yeast mutants. Among the several genes that we discovered is MGR1, which encodes a novel subunit of the i AAA protease complex located in the mitochondrial inner membrane. mgr1Delta mutants retain some i-AAA protease activity, yet mitochondria lacking Mgr1p contain a misassembled i-AAA protease and are defective for turnover of mitochondrial inner membrane proteins. Our results highlight the importance of the i-AAA complex and proteolysis at the inner membrane in cells lacking mitochondrial DNA. PMID- 16267277 TI - Characterization of an ERAD gene as VPS30/ATG6 reveals two alternative and functionally distinct protein quality control pathways: one for soluble Z variant of human alpha-1 proteinase inhibitor (A1PiZ) and another for aggregates of A1PiZ. AB - The Z variant of human alpha-1 proteinase inhibitor (A1PiZ) is a substrate for endoplasmic reticulum-associated protein degradation (ERAD). To identify genes required for the degradation of this protein, A1PiZ degradation-deficient (add) yeast mutants were isolated. The defect in one of these mutants, add3, was complemented by VPS30/ATG6, a gene that encodes a component of two phosphatidylinositol 3-kinase (PtdIns 3-kinase) complexes: complex I is required for autophagy, whereas complex II is required for the carboxypeptidase Y (CPY)-to vacuole pathway. We found that upon overexpression of A1PiZ, both PtdIns 3-kinase complexes were required for delivery of the excess A1PiZ to the vacuole. When the CPY-to-vacuole pathway was compromised, A1PiZ was secreted; however, disruption of autophagy led to an increase in aggregated A1PiZ rather than secretion. These results suggest that excess soluble A1PiZ transits the secretion pathway to the trans-Golgi network and is selectively targeted to the vacuole via the CPY-to vacuole sorting pathway, but excess A1PiZ that forms aggregates in the endoplasmic reticulum is targeted to the vacuole via autophagy. These findings illustrate the complex nature of protein quality control in the secretion pathway and reveal multiple sites that recognize and sort both soluble and aggregated forms of aberrant or misfolded proteins. PMID- 16267276 TI - The Flo8 transcription factor is essential for hyphal development and virulence in Candida albicans. AB - The transcription factor Flo8 is essential for filamentous growth in Saccharomyces cerevisiae and is regulated under the cAMP/protein kinase A (PKA) pathway. To determine whether a similar pathway/regulation exists in Candida albicans, we have cloned C. albicans FLO8 by its ability to complement S. cerevisiae flo8. Deleting FLO8 in C. albicans blocked hyphal development and hypha-specific gene expression. The flo8/flo8 mutant is avirulent in a mouse model of systemic infection. Genome-wide transcription profiling of efg1/efg1 and flo8/flo8 using a C. albicans DNA microarray suggests that Flo8 controls subsets of Efg1-regulated genes. Most of these genes are hypha specific, including HGC1 and IHD1. We also show that Flo8 interacts with Efg1 in yeast and hyphal cells by in vivo immunoprecipitation. Similar to efg1/efg1, flo8/flo8 and cdc35/cdc35 show enhanced hyphal growth under an embedded growth condition. Our results suggest that Flo8 may function downstream of the cAMP/PKA pathway, and together with Efg1, regulates the expression of hypha-specific genes and genes that are important for the virulence of C. albicans. PMID- 16267278 TI - Escherichia coli starvation diets: essential nutrients weigh in distinctly. PMID- 16267279 TI - Contributions of ATP, GTP, and redox state to nutritional stress activation of the Bacillus subtilis sigmaB transcription factor. AB - The general stress regulon of Bacillus subtilis is induced by activation of the sigma(B) transcription factor. sigma(B) activation occurs when one of two phosphatases responds to physical or nutritional stress to activate a positive sigma(B) regulator by dephosphorylation. The signal that triggers the nutritional stress phosphatase (RsbP) is unknown; however, RsbP activation occurs under culture conditions (glucose/phosphate starvation, azide or decoyinine treatment) that reduce the cell's levels of ATP and/or GTP. Variances in nucleotide levels in these instances may be coincidental rather than causal. RsbP carries a domain (PAS) that in some regulatory systems can respond directly to changes in electron transport, proton motive force, or redox potential, changes that typically precede shifts in high-energy nucleotide levels. The current work uses Bacillus subtilis with mutations in the oxidative phosphorylation and purine nucleotide biosynthetic pathways in conjunction with metabolic inhibitors to better define the inducing signal for RsbP activation. The data argue that a drop in ATP, rather than changes in GTP, proton motive force, or redox state, is the key to triggering sigma(B) activation. PMID- 16267280 TI - Adaptations of Pseudomonas aeruginosa to the cystic fibrosis lung environment can include deregulation of zwf, encoding glucose-6-phosphate dehydrogenase. AB - Cystic fibrosis (CF) patients are highly susceptible to chronic pulmonary disease caused by mucoid Pseudomonas aeruginosa strains that overproduce the exopolysaccharide alginate. We showed here that a mutation in zwf, encoding glucose-6-phosphate dehydrogenase (G6PDH), leads to a approximately 90% reduction in alginate production in the mucoid, CF isolate, P. aeruginosa FRD1. The main regulator of alginate, sigma-22 encoded by algT (algU), plays a small but demonstrable role in the induction of zwf expression in P. aeruginosa. However, G6PDH activity and zwf expression were higher in FRD1 strains than in PAO1 strains. In PAO1, zwf expression and G6PDH activity are known to be subject to catabolite repression by succinate. In contrast, FRD1 zwf expression and G6PDH activity were shown to be refractory to such catabolite repression. This was apparently not due to a defect in the catabolite repression control (Crc) protein. Such relaxed control of zwf was found to be common among several examined CF isolates but was not seen in other strains of clinical and environmental origin. Two sets of clonal isolates from individual CF patient indicated that the resident P. aeruginosa strain underwent an adaptive change that deregulated zwf expression. We hypothesized that high-level, unregulated G6PDH activity provided a survival advantage to P. aeruginosa within the lung environment. Interestingly, zwf expression in P. aeruginosa was shown to be required for its resistance to human sputum. This study illustrates that adaptation to the CF pulmonary environment by P. aeruginosa can include altered regulation of basic metabolic activities, including carbon catabolism. PMID- 16267281 TI - Unconventional mode of attachment of the Ruminococcus flavefaciens cellulosome to the cell surface. AB - Sequence extension of the scaffoldin gene cluster from Ruminococcus flavefaciens revealed a new gene (scaE) that encodes a protein with an N-terminal cohesin domain and a C terminus with a typical gram-positive anchoring signal for sortase mediated attachment to the bacterial cell wall. The recombinant cohesin of ScaE was recovered after expression in Escherichia coli and was shown to bind to the C terminal domain of the cellulosomal structural protein ScaB, as well as to three unknown polypeptides derived from native cellulose-bound Ruminococcus flavefaciens protein extracts. The ScaB C terminus includes a cryptic dockerin domain that is unusual in its sequence, and considerably larger than conventional dockerins. The ScaB dockerin binds to ScaE, suggesting that this interaction occurs through a novel cohesin-dockerin pairing. The novel ScaB dockerin was expressed as a xylanase fusion protein, which was shown to bind tenaciously and selectively to a recombinant form of the ScaE cohesin. Thus, ScaE appears to play a role in anchoring the cellulosomal complex to the bacterial cell envelope via its interaction with ScaB. This sortase-mediated mechanism for covalent cell-wall anchoring of the cellulosome in R. flavefaciens differs from those reported thus far for any other cellulosome system. PMID- 16267282 TI - Adenylate cyclase toxin (ACT) from Bordetella hinzii: characterization and differences from ACT of Bordetella pertussis. AB - Bordetella hinzii is a commensal respiratory microorganism in poultry but is increasingly being recognized as an opportunistic pathogen in immunocompromised humans. Although associated with a variety of disease states, practically nothing is known about the mechanisms employed by this bacterium. In this study, we show by DNA sequencing and reverse transcription-PCR that both commensal and clinical strains of B. hinzii possess and transcriptionally express cyaA, the gene encoding adenylate cyclase toxin (ACT) in other pathogenic Bordetella species. By Western blotting, we also found that B. hinzii produces full-length ACT protein in quantities that are comparable to those made by B. pertussis. In contrast to B. pertussis ACT, however, ACT from B. hinzii is less extractable from whole bacteria, nonhemolytic, has a 50-fold reduction in adenylate cyclase activity, and is unable to elevate cyclic AMP levels in host macrophages (nontoxic). The decrease in enzymatic activity is attributable, at least in part, to a decreased binding affinity of B. hinzii ACT for calmodulin, the eukaryotic activator of B. pertussis ACT. In addition, we demonstrate that the lack of intoxication by B. hinzii ACT may be due to the absence of expression of cyaC, the gene encoding the accessory protein required for the acylation of B. pertussis ACT. These results demonstrate the expression of ACT by B. hinzii and represent the first characterization of a potential virulence factor of this organism. PMID- 16267283 TI - The activity profile of the NhaD-type Na+(Li+)/H+ antiporter from the soda Lake Haloalkaliphile Alkalimonas amylolytica is adaptive for the extreme environment. AB - In extreme alkaliphiles, Na(+)/H(+) antiporters play a central role in the Na(+) cycle that supports pH homeostasis, Na(+) resistance, solute uptake, and motility. Properties of individual antiporters have only been examined in extremely alkaliphilic soil Bacillus spp., whereas the most alkaline natural habitats usually couple high pH with high salinity. Here, studies were conducted on a Na(+)(Li(+))/H(+) antiporter, NhaD, from the soda lake haloalkaliphile Alkalimonas amylolytica. The activity profile of A. amylolytica NhaD at different pH values and Na(+) concentrations reflects its unique natural habitat. In membrane vesicles from antiporter-deficient Escherichia coli EP432 (DeltanhaA DeltanhaB), the pH optimum for NhaD-dependent Na(+)(Li(+))/H(+) antiport was at least 9.5, the highest pH that could be tested; no activity was observed at pH < or =8.5. NhaD supported low Na(+)/H(+) antiport activity at pH 9.5 that was detectable over a range of Na(+) concentrations from 10 mM to at least 800 mM, with a 600 mM optimum. Although A. amylolytica nhaD was isolated by complementing the Li(+) sensitivity of the triple mutant E. coli strain KNabc (DeltanhaA DeltanhaB DeltachaA), sustained propagation of nhaD-bearing plasmids in this strain resulted in a glycine (Gly(327))-->serine mutation in a putative cytoplasmic loop of the mutant transporter. The altered activity profile of NhaD G327S appears to be adaptive to the E. coli setting: a much higher activity than wild-type NhaD at Na(+) concentrations up to 200 mM but lower activity at 400 to 600 mM Na(+), with a pH optimum and minimal pH for activity lower than those of wild-type NhaD. PMID- 16267284 TI - Conditional depletion of KasA, a key enzyme of mycolic acid biosynthesis, leads to mycobacterial cell lysis. AB - Inhibition or inactivation of InhA, a fatty acid synthase II (FASII) enzyme, leads to mycobacterial cell lysis. To determine whether inactivation of other enzymes of the mycolic acid-synthesizing FASII complex also leads to lysis, we characterized the essentiality of two beta-ketoacyl-acyl carrier protein synthases, KasA and KasB, in Mycobacterium smegmatis. Using specialized transduction for allelic exchange, null kasB mutants, but not kasA mutants, could be generated in Mycobacterium smegmatis, suggesting that unlike kasB, kasA is essential. To confirm the essentiality of kasA, and to detail the molecular events that occur following depletion of KasA, we developed CESTET (conditional expression specialized transduction essentiality test), a genetic tool that combines conditional gene expression and specialized transduction. Using CESTET, we were able to generate conditional null inhA and kasA mutants. We studied the effects of depletion of KasA in M. smegmatis using the former strain as a reference. Depletion of either InhA or KasA led to cell lysis, but with different biochemical and morphological events prior to lysis. While InhA depletion led to the induction of an 80-kDa complex containing both KasA and AcpM, the mycobacterial acyl carrier protein, KasA depletion did not induce the same complex. Depletion of either InhA or KasA led to inhibition of alpha and epoxy mycolate biosynthesis and to accumulation of alpha'-mycolates. Furthermore, scanning electron micrographs revealed that KasA depletion resulted in the cell surface having a "crumpled" appearance, in contrast to the blebs observed on InhA depletion. Thus, our studies support the further exploration of KasA as a target for mycobacterial-drug development. PMID- 16267285 TI - Roles of the Escherichia coli RecA protein and the global SOS response in effecting DNA polymerase selection in vivo. AB - The Escherichia coli beta sliding clamp protein is proposed to play an important role in effecting switches between different DNA polymerases during replication, repair, and translesion DNA synthesis. We recently described how strains bearing the dnaN159 allele, which encodes a mutant form of the beta clamp (beta159), display a UV-sensitive phenotype that is suppressed by inactivation of DNA polymerase IV (M. D. Sutton, J. Bacteriol. 186:6738-6748, 2004). As part of an ongoing effort to understand mechanisms of DNA polymerase management in E. coli, we have further characterized effects of the dnaN159 allele on polymerase usage. Three of the five E.coli DNA polymerases (II, IV, and V) are regulated as part of the global SOS response. Our results indicate that elevated expression of the dinB-encoded polymerase IV is sufficient to result in conditional lethality of the dnaN159 strain. In contrast, chronically activated RecA protein, expressed from the recA730 allele, is lethal to the dnaN159 strain, and this lethality is suppressed by mutations that either mitigate RecA730 activity (i.e., DeltarecR), or impair the activities of DNA polymerase II or DNA polymerase V (i.e., DeltapolB or DeltaumuDC). Thus, we have identified distinct genetic requirements whereby each of the three different SOS-regulated DNA polymerases are able to confer lethality upon the dnaN159 strain, suggesting the presence of multiple mechanisms by which the actions of the cell's different DNA polymerases are managed in vivo. PMID- 16267286 TI - High-resolution visualization of Pseudomonas aeruginosa PAO1 biofilms by freeze substitution transmission electron microscopy. AB - High-pressure freeze-substitution and transmission electron microscopy have been used for high-resolution imaging of the natural structure of a gram-negative biofilm. Unlike more conventional embedding techniques, this method confirms many of the observations seen by confocal microscopy but with finer structural detail. It further reveals that there is a structural complexity to biofilms at both the cellular and extracellular matrix levels that has not been seen before. Different domains of healthy and lysed cells exist randomly dispersed within a single biofilm as well as different structural organizations of exopolymers. Particulate matter is suspended within this network of fibers and appears to be an integral part of the exopolymeric substance (EPS). O-side chains extending from the outer membrane are integrated into EPS polymers so as to form a continuum. Together, the results support the concept of physical microenvironments within biofilms and show a complexity that was hitherto unknown. PMID- 16267287 TI - Bacillus subtilis cysteine synthetase is a global regulator of the expression of genes involved in sulfur assimilation. AB - The synthesis of L-cysteine, the major mechanism by which sulfur is incorporated into organic compounds in microorganisms, occupies a significant fraction of bacterial metabolism. In Bacillus subtilis the cysH operon, encoding several proteins involved in cysteine biosynthesis, is induced by sulfur starvation and tightly repressed by cysteine. We show that a null mutation in the cysK gene encoding an O-acetylserine-(thiol)lyase, the enzyme that catalyzes the final step in cysteine biosynthesis, results in constitutive expression of the cysH operon. Using DNA microarrays we found that, in addition to cysH, almost all of the genes required for sulfate assimilation are constitutively expressed in cysK mutants. These results indicate that CysK, besides its enzymatic role in cysteine biosynthesis, is a global negative regulator of genes involved in sulfur metabolism. PMID- 16267288 TI - Functional analysis of all aminotransferase proteins inferred from the genome sequence of Corynebacterium glutamicum. AB - Twenty putative aminotransferase (AT) proteins of Corynebacterium glutamicum, or rather pyridoxal-5'-phosphate (PLP)-dependent enzymes, were isolated and assayed among others with L-glutamate, L-aspartate, and L-alanine as amino donors and a number of 2-oxo-acids as amino acceptors. One outstanding AT identified is AlaT, which has a broad amino donor specificity utilizing (in the order of preference) L-glutamate > 2-aminobutyrate > L-aspartate with pyruvate as acceptor. Another AT is AvtA, which utilizes L-alanine to aminate 2-oxo-isovalerate, the L-valine precursor, and 2-oxo-butyrate. A second AT active with the L-valine precursor and that of the other two branched-chain amino acids, too, is IlvE, and both enzyme activities overlap partially in vivo, as demonstrated by the analysis of deletion mutants. Also identified was AroT, the aromatic AT, and this and IlvE were shown to have comparable activities with phenylpyruvate, thus demonstrating the relevance of both ATs for L-phenylalanine synthesis. We also assessed the activity of two PLP-containing cysteine desulfurases, supplying a persulfide intermediate. One of them is SufS, which assists in the sulfur transfer pathway for the Fe-S cluster assembly. Together with the identification of further ATs and the additional analysis of deletion mutants, this results in an overview of the ATs within an organism that may not have been achieved thus far. PMID- 16267289 TI - Stabilization of polar localization of a chemoreceptor via its covalent modifications and its communication with a different chemoreceptor. AB - In the chemotaxis of Escherichia coli, polar clustering of the chemoreceptors, the histidine kinase CheA, and the adaptor protein CheW is thought to be involved in signal amplification and adaptation. However, the mechanism that leads to the polar localization of the receptor is still largely unknown. In this study, we examined the effect of receptor covalent modification on the polar localization of the aspartate chemoreceptor Tar fused to green fluorescent protein (GFP). Amidation (and presumably methylation) of Tar-GFP enhanced its own polar localization, although the effect was small. The slight but significant effect of amidation on receptor localization was reinforced by the fact that localization of a noncatalytic mutant version of GFP-CheR that targets to the C-terminal pentapeptide sequence of Tar was similarly facilitated by receptor amidation. Polar localization of the demethylated version of Tar-GFP was also enhanced by increasing levels of the serine chemoreceptor Tsr. The effect of covalent modification on receptor localization by itself may be too small to account for chemotactic adaptation, but receptor modification is suggested to contribute to the molecular assembly of the chemoreceptor/histidine kinase array at a cell pole, presumably by stabilizing the receptor dimer-to-dimer interaction. PMID- 16267290 TI - Genetic composition of the Bacillus subtilis SOS system. AB - The SOS response in bacteria includes a global transcriptional response to DNA damage. DNA damage is sensed by the highly conserved recombination protein RecA, which facilitates inactivation of the transcriptional repressor LexA. Inactivation of LexA causes induction (derepression) of genes of the LexA regulon, many of which are involved in DNA repair and survival after DNA damage. To identify potential RecA-LexA-regulated genes in Bacillus subtilis, we searched the genome for putative LexA binding sites within 300 bp upstream of the start codons of all annotated open reading frames. We found 62 genes that could be regulated by putative LexA binding sites. Using mobility shift assays, we found that LexA binds specifically to DNA in the regulatory regions of 54 of these genes, which are organized in 34 putative operons. Using DNA microarray analyses, we found that 33 of the genes with LexA binding sites exhibit RecA-dependent induction by both mitomycin C and UV radiation. Among these 33 SOS genes, there are 22 distinct LexA binding sites preceding 18 putative operons. Alignment of the distinct LexA binding sites reveals an expanded consensus sequence for the B. subtilis operator: 5'-CGAACATATGTTCG-3'. Although the number of genes controlled by RecA and LexA in B. subtilis is similar to that of Escherichia coli, only eight B. subtilis RecA-dependent SOS genes have homologous counterparts in E. coli. PMID- 16267291 TI - The twin-arginine translocation pathway of Mycobacterium smegmatis is functional and required for the export of mycobacterial beta-lactamases. AB - The twin-arginine translocation (Tat) pathway exports folded proteins across the bacterial cytoplasmic membrane and is responsible for the proper extracytoplasmic localization of proteins involved in a variety of cellular functions, including pathogenesis. The Mycobacterium tuberculosis and Mycobacterium smegmatis genomes contain open reading frames with homology to components of the Tat export system (TatABC) as well as potential Tat-exported proteins possessing N-terminal signal sequences with the characteristic twin-arginine motif. Due to the importance of exported virulence factors in the pathogenesis of M. tuberculosis and the limited understanding of mycobacterial protein export systems, we sought to determine the functional nature of the Tat export pathway in mycobacteria. Here we describe phenotypic analyses of DeltatatA and DeltatatC deletion mutants of M. smegmatis, which demonstrated that tatA and tatC encode components of a functional Tat system capable of exporting characteristic Tat substrates. Both mutants displayed a growth defect on agar medium and hypersensitivity to sodium dodecyl sulfate. The mutants were also defective in the export of active beta-lactamases of M. smegmatis (BlaS) and M. tuberculosis (BlaC), both of which possess twin-arginine signal sequences. The Tat-dependent nature of BlaC was further revealed by mutation of the twin-arginine motif. Finally, we demonstrated that replacement of the native signal sequence of BlaC with the predicted Tat signal sequences of M. tuberculosis phospholipase C proteins (PlcA and PlcB) resulted in the Tat dependent export of an enzymatically active 'BlaC. Thus, 'BlaC can be used as a genetic reporter for Tat-dependent export in mycobacteria. PMID- 16267292 TI - Periplasmic peptidyl prolyl cis-trans isomerases are not essential for viability, but SurA is required for pilus biogenesis in Escherichia coli. AB - In Escherichia coli, FkpA, PpiA, PpiD, and SurA are the four known periplasmic cis-trans prolyl isomerases. These isomerases facilitate proper protein folding by increasing the rate of transition of proline residues between the cis and trans states. Genetic inactivation of all four periplasmic isomerases resulted in a viable strain that exhibited a decreased growth rate and increased susceptibility to certain antibiotics. Levels of the outer membrane proteins LamB and OmpA in the quadruple mutant were indistinguishable from those in the surA single mutant. In addition, expression of P and type 1 pili (adhesive organelles produced by uropathogenic strains of E. coli and assembled by the chaperone/usher pathway) were severely diminished in the absence of the four periplasmic isomerases. Maturation of the usher was significantly impaired in the outer membranes of strains devoid of all four periplasmic isomerases, resulting in a defect in pilus assembly. Moreover, this defect in pilus assembly and usher stability could be attributed to the absence of SurA. The data presented here suggest that the four periplasmic isomerases are not essential for growth under laboratory conditions but may have significant roles in survival in environmental and pathogenic niches, as indicated by the effect on pilus production. PMID- 16267293 TI - Diverse phenotypes resulting from polyphosphate kinase gene (ppk1) inactivation in different strains of Helicobacter pylori. AB - Connections among biochemical pathways should help buffer organisms against environmental stress and affect the pace and trajectory of genome evolution. To explore these ideas, we studied consequences of inactivating the gene for polyphosphate kinase 1 (ppk1) in strains of Helicobacter pylori, a genetically diverse gastric pathogen. The PPK1 enzyme catalyzes synthesis of inorganic polyphosphate (poly P), a reservoir of high-energy phosphate bonds with multiple roles. Prior analyses in less-fastidious microbes had implicated poly P in stress resistance, motility, and virulence. In our studies, ppk1 inactivation caused the expected near-complete absence of poly P (>250-fold decrease) but had phenotypic effects that differed markedly among unrelated strains: (i) poor initial growth on standard brain heart infusion agar (five of six strains tested); (ii) weakened colonization of mice (4 of 5 strains); (iii) reduced growth on Ham's F-12 agar, a nutritionally limiting medium (8 of 11 strains); (iv) heightened susceptibility to metronidazole (6 of 17 strains); and (v) decreased motility in soft agar (1 of 13 strains). Complementation tests confirmed that the lack of growth of one Deltappk1 strain on F-12 agar and the inability to colonize mice of another were each due to ppk1 inactivation. Thus, the importance of ppk1 to H. pylori differed among strains and the phenotypes monitored. We suggest that quantitative interactions, as seen here, are common among genes that affect metabolic pathways and that H. pylori's high genetic diversity makes it well suited for studies of such interactions, their underlying mechanisms, and their evolutionary consequences. PMID- 16267294 TI - Molecular and biochemical characterization of the xlnD-encoded 3-hydroxybenzoate 6-hydroxylase involved in the degradation of 2,5-xylenol via the gentisate pathway in Pseudomonas alcaligenes NCIMB 9867. AB - The xlnD gene from Pseudomonas alcaligenes NCIMB 9867 (strain P25X) was shown to encode 3-hydroxybenzoate 6-hydroxylase I, the enzyme that catalyzes the NADH dependent conversion of 3-hydroxybenzoate to gentisate. Active recombinant XlnD was purified as a hexahistidine fusion protein from Escherichia coli, had an estimated molecular mass of 130 kDa, and is probably a trimeric protein with a subunit mass of 43 kDa. This is in contrast to the monomeric nature of the few 3 hydroxybenzoate 6-hydroxylases that have been characterized thus far. Like other 3-hydroxybenzoate 6-hydroxylases, XlnD could utilize either NADH or NADPH as the electron donor. P25X harbors a second 3-hydroxybenzoate 6-hydroxylase II that was strictly inducible by specific aromatic substrates. However, the degradation of 2,5-xylenol and 3,5-xylenol in strain P25X was found to be dependent on the xlnD encoded 6-hydroxylase I and not the second, strictly inducible 6-hydroxylase II. PMID- 16267295 TI - In vitro analysis of protein-operator interactions of the NikR and fur metal responsive regulators of coregulated genes in Helicobacter pylori. AB - Two important metal-responsive regulators, NikR and Fur, are involved in nickel and iron homeostasis and controlling gene expression in Helicobacter pylori. To date, they have been implicated in the regulation of sets of overlapping genes. We have attempted here dissection of the molecular mechanisms involved in transcriptional regulation of the NikR and Fur proteins, and we investigated protein-promoter interactions of the regulators with known target genes. We show that H. pylori NikR is a tetrameric protein and, through DNase I footprinting analysis, we have identified operators for NikR to which it binds with different affinities in a metal-responsive way. Mapping of the NikR binding site upstream of the urease promoter established a direct role for NikR as a positive regulator of transcription and, through scanning mutagenesis of this binding site, we have determined two subsites that are important for the binding of the protein to its target sequence. Furthermore, by alignment of the operators for NikR, we have shown that the H. pylori protein recognizes a sequence that is distinct from its well-studied orthologue in Escherichia coli. Moreover, we show that NikR and Fur can bind independently at distinct operators and also compete for overlapping operators in some coregulated gene promoters, adding another dimension to the previous suggested link between iron and nickel regulation. Finally, the importance of an interconnection between metal-responsive gene networks for homeostasis is discussed. PMID- 16267296 TI - Evidence for acquisition of Legionella type IV secretion substrates via interdomain horizontal gene transfer. AB - Intracellular pathogens exploit host cell functions to create a replication niche inside eukaryotic cells. The causative agent of Legionnaires' disease, the gamma proteobacterium Legionella pneumophila, resides and replicates within a modified vacuole of protozoan and mammalian cells. L. pneumophila translocates effector proteins into host cells through the Icm-Dot complex, a specialized type IVB secretion system that is required for intracellular growth. To find out if some effector proteins may have been acquired through interdomain horizontal gene transfer (HGT), we performed a bioinformatic screen that searched for eukaryotic motifs in all open reading frames of the L. pneumophila Philadelphia-1 genome. We found 44 uncharacterized genes with many distinct eukaryotic motifs. Most of these genes contain G+C biases compared to other L. pneumophila genes, supporting the theory that they were acquired through HGT. Furthermore, we found that several of them are expressed and up-regulated in stationary phase in an RpoS dependent manner. In addition, at least seven of these gene products are translocated into host cells via the Icm-Dot complex, confirming their role in the intracellular environment. Reminiscent of the case with most Icm-Dot substrates, most of the strains containing mutations in these genes grew comparably to the parent strain intracellularly. Our findings suggest that in L. pneumophila, interdomain HGT may have been a major mechanism for the acquisition of determinants of infection. PMID- 16267297 TI - Genetic analysis of transfer-related regions of the vancomycin resistance Enterococcus conjugative plasmid pHTbeta: identification of oriT and a putative relaxase gene. AB - The pHT plasmids pHTalpha (65.9 kbp), pHTbeta (63.7 kbp), and pHTgamma (66.5 kbp) are highly conjugative pheromone-independent pMG1-like plasmids that carry Tn1546 like transposons encoding vancomycin resistance. pHTbeta is the prototype plasmid, and the pHTalpha and pHTgamma plasmids are derivatives of the insertion into pHTbeta of an IS232-like (2.2 kbp) element and a group II intron (2.8 kbp), respectively. The complete nucleotide sequence of the pHTbeta plasmid was determined and, with the exception of the Tn1546-like insertion (10,851 bp), was found to be 52,890 bp. Sixty-one open reading frames (ORFs) having the same transcript orientation were identified. A homology search revealed that 22 of the pHTbeta (pHT) plasmid ORFs showed similarities to the ORFs identified on the pXO2 plasmid (96.2 kbp), which is the virulence plasmid essential for capsule formation by Bacillus anthracis; however, the functions of most of the ORFs remain unknown. Most other ORFs did not show any significant homology to reported genes for which functions have been analyzed. To investigate the highly efficient transfer mechanism of the pHT plasmid, mutations with 174 unique insertions of transposon Tn917-lac insertion mutants of pHTbeta were obtained. Of the 174 derivatives, 92 showed decrease or loss in transfer frequency, and 74 showed normal transfer frequency and LacZ expression. Eight derivatives showed normal transfer and no LacZ expression. Inserts within the 174 derivatives were mapped to 124 different sites on pHTbeta. The Tn917-lac insertions which resulted in altered transfer frequency mapped to three separate regions designated I, II, and III, which were separated by segments in which insertions of Tn917-lac did not affect transfer. There was no region homologous to the previously reported oriT sequences in the pHT plasmid. The oriT was cloned by selection for the ability to mobilize the vector plasmid pAM401. The oriT region resided in a noncoding region (192 bp) between ORF31 and ORF32 and contained three direct repeat sequences and two inverted repeat sequences. ORF34, encoding a 506-amino-acid protein which was located downstream of the oriT region, contains the three conserved motifs (I to III) of the DNA relaxase/nickase of mobile plasmids. The transfer abilities of the Tn917-lac-insertion mutants of ORF34 or a mutant of ORF34 with an in-frame motif III deletion were completely abolished. The sequence of the oriT region and the deduced relaxase/nickase protein of ORF34 showed no significant similarity to the oriT and relaxase/nickase of other conjugative plasmids, respectively. The putative relaxase/nickase protein of ORF34 could be classified as a new member of the MOB(MG) family. PMID- 16267299 TI - Assembly and function of a spore coat-associated transglutaminase of Bacillus subtilis. AB - The assembly of a multiprotein coat around the Bacillus subtilis spore confers resistance to lytic enzymes and noxious chemicals and ensures normal germination. Part of the coat is cross-linked and resistant to solubilization. The coat contains epsilon-(gamma-glutamyl)lysyl cross-links, and the expression of the gene (tgl) for a spore-associated transglutaminase was shown before to be required for the cross-linking of coat protein GerQ. Here, we have investigated the assembly and function of Tgl. We found that Tgl associates, albeit at somewhat reduced levels, with the coats of mutants that are unable to assemble the outer coat (cotE), that are missing the inner coat and with a greatly altered outer coat (gerE), or that are lacking discernible inner and outer coat structures (cotE gerE double mutant). This suggests that Tgl is present at various levels within the coat lattice. The assembly of Tgl occurs independently of its own activity, as a single amino acid substitution of a cysteine to an alanine (C116A) at the active site of Tgl does not affect its accumulation or assembly. However, like a tgl insertional mutation, the tglC116A allele causes increased extractability of polypeptides of about 40, 28, and 16 kDa in addition to GerQ (20 kDa) and affects the structural integrity of the coat. We show that most Tgl is assembled onto the spore surface soon after its synthesis in the mother cell under sigma(K) control but that the complete insolubilization of at least two of the Tgl-controlled polypeptides occurs several hours later. We also show that a multicopy allele of tgl causes increased assembly of Tgl and affects the assembly, structure, and functional properties of the coat. PMID- 16267298 TI - Mapping of a YscY binding domain within the LcrH chaperone that is required for regulation of Yersinia type III secretion. AB - Type III secretion systems are used by many animal and plant interacting bacteria to colonize their host. These systems are often composed of at least 40 genes, making their temporal and spatial regulation very complex. Some type III chaperones of the translocator class are important regulatory molecules, such as the LcrH chaperone of Yersinia pseudotuberculosis. In contrast, the highly homologous PcrH chaperone has no regulatory effect in native Pseudomonas aeruginosa or when produced in Yersinia. In this study, we used LcrH-PcrH chaperone hybrids to identify a discrete region in the N terminus of LcrH that is necessary for YscY binding and regulatory control of the Yersinia type III secretion machinery. PcrH was unable to bind YscY and the homologue Pcr4 of P. aeruginosa. YscY and Pcr4 were both essential for type III secretion and reciprocally bound to both substrates YscX of Yersinia and Pcr3 of P. aeruginosa. Still, Pcr4 was unable to complement a DeltayscY null mutant defective for type III secretion and yop-regulatory control in Yersinia, despite the ability of YscY to function in P. aeruginosa. Taken together, we conclude that the cross-talk between the LcrH and YscY components represents a strategic regulatory pathway specific to Yersinia type III secretion. PMID- 16267300 TI - CapE, a 47-amino-acid peptide, is necessary for Bacillus anthracis polyglutamate capsule synthesis. AB - Polyglutamate is found in various bacteria, but displays different functions depending on the species and their environment. Here, we describe a minimal polyglutamate synthesis system in Bacillus anthracis. In addition to the three genes previously described as sufficient for polyglutamate synthesis, this system includes a small open reading frame, capE, belonging to the cap operon. The polyglutamate system's requirement for the five cap genes, for capsulation and anchoring, was assayed in nonpolar mutants. The capA, capB, capC, and capE genes are all necessary and are sufficient for polyglutamate synthesis by B. anthracis. capD is required for polyglutamate anchoring to the peptidoglycan. The 47-amino acid peptide encoded by capE is localized in the B. anthracis membrane. It is not a regulator and it is required for polyglutamate synthesis, suggesting that it has a structural role in polyglutamate synthesis. CapE appears to interact with CapA. Bacillus subtilis ywtC is similar to capE and we named it pgsE. Genes similar to capE or pgsE were found in B. subtilis natto, Bacillus licheniformis, and Staphylococcus epidermidis, species that produce polyglutamate. All the bacterial polyglutamate synthesis systems analyzed show a similar genetic organization and, we suggest, the same protein requirements. PMID- 16267302 TI - Functional genomic analysis of three nitrogenase isozymes in the photosynthetic bacterium Rhodopseudomonas palustris. AB - The photosynthetic bacterium Rhodopseudomonas palustris is one of just a few prokaryotes described so far that has vnf and anf genes for alternative vanadium cofactor (V) and iron cofactor (Fe) nitrogenases in addition to nif genes for a molybdenum cofactor (Mo) nitrogenase. Transcriptome data indicated that the 32 genes in the nif gene cluster, but not the anf or vnf genes, were induced in wild type and Mo nitrogenase-expressing strains grown under nitrogen-fixing conditions in Mo-containing medium. Strains that were unable to express a functional Mo nitrogenase due to mutations in Mo nitrogenase structural genes synthesized functional V and Fe nitrogenases and expressed vnf and anf genes in nitrogen fixing growth media that contained Mo and V at concentrations far in excess of those that repress alternative nitrogenase gene expression in other bacteria. Thus, not only does R. palustris have multiple enzymatic options for nitrogen fixation, but in contrast to reports on other nitrogen-fixing bacteria, the expression of its alternative nitrogenases is not repressed by transition metals. Between 95 and 295 genes that are not directly associated with nitrogenase synthesis and assembly were induced under nitrogen-fixing conditions, depending on which nitrogenase was being used by R. palustris. Genes for nitrogen acquisition were expressed at particularly high levels during alternative nitrogenase-dependent growth. This suggests that alternative nitrogenase expressing cells are relatively starved for nitrogen and raises the possibility that fixed nitrogen availability may be the primary signal that controls the synthesis of the V and Fe nitrogenases. PMID- 16267301 TI - A gyrase mutant with low activity disrupts supercoiling at the replication terminus. AB - When a mutation in an essential gene shows a temperature-sensitive phenotype, one usually assumes that the protein is inactive at nonpermissive temperature. DNA gyrase is an essential bacterial enzyme composed of two subunits, GyrA and GyrB. The gyrB652 mutation results from a single base change that substitutes a serine residue for arginine 436 (R436-S) in the GyrB protein. At 42 degrees C, strains with the gyrB652 allele stop DNA replication, and at 37 degrees C, such strains grow but have RecA-dependent SOS induction and show constitutive RecBCD-dependent DNA degradation. Surprisingly, the GyrB652 protein is not inactive at 42 degrees C in vivo or in vitro and it doesn't directly produce breaks in chromosomal DNA. Rather, this mutant has a low k(cat) compared to wild-type GyrB subunit. With more than twice the normal mean number of supercoil domains, this gyrase hypomorph is prone to fork collapse and topological chaos near the terminus of DNA replication. PMID- 16267303 TI - Characterization of Mycobacterium tuberculosis Rv3676 (CRPMt), a cyclic AMP receptor protein-like DNA binding protein. AB - Little is known about cyclic AMP (cAMP) function in Mycobacterium tuberculosis, despite its ability to encode 15 adenylate cyclases and 10 cNMP-binding proteins. M. tuberculosis Rv3676, which we have designated CRP(Mt), is predicted to be a cAMP-dependent transcription factor. In this study, we characterized CRP(Mt)'s interactions with DNA and cAMP, using experimental and computational approaches. We used Gibbs sampling to define a CRP(Mt) DNA motif that resembles the cAMP receptor protein (CRP) binding motif model for Escherichia coli. CRP(Mt) binding sites were identified in a total of 73 promoter regions regulating 114 genes in the M. tuberculosis genome, which are being explored as a regulon. Specific CRP(Mt) binding caused DNA bending, and the substitution of highly conserved nucleotides in the binding site resulted in a complete loss of binding to CRP(Mt). cAMP enhanced CRP(Mt)'s ability to bind DNA and caused allosteric alterations in CRP(Mt) conformation. These results provide the first direct evidence for cAMP binding to a transcription factor in M. tuberculosis, suggesting a role for cAMP signal transduction in M. tuberculosis and implicating CRP(Mt) as a cAMP-responsive global regulator. PMID- 16267304 TI - Novel virulence gene of Pseudomonas syringae pv. tomato strain DC3000. AB - Previously, we conducted a mutant screen of Pseudomonas syringae pv. tomato strain DC3000 to identify genes that contribute to virulence on Arabidopsis thaliana plants. Here we describe the characterization of one mutant strain, DB4H2, which contains a single Tn5 insertion in PSPTO3576, an open reading frame that is predicted to encode a protein belonging to the TetR family of transcriptional regulators. We demonstrate that PSPTO3576 is necessary for virulence in DC3000 and designate the encoded protein TvrR (TetR-like virulence regulator). TvrR, like many other TetR-like transcriptional regulators, negatively regulates its own expression. Despite the presence of a putative HrpL binding site in the tvrR promoter region, tvrR is not regulated by HrpL, an alternative sigma factor that regulates the expression of many known DC3000 virulence genes. tvrR mutant strains grow comparably to wild-type DC3000 in culture and possess an intact type III secretion system. However, tvrR mutants do not cause disease symptoms on inoculated A. thaliana and tomato plants, and their growth within plant tissue is significantly impaired. We demonstrate that tvrR mutant strains are able to synthesize coronatine (COR), a phytotoxin required for virulence of DC3000 on A. thaliana. Given that tvrR mutant strains are not defective for type III secretion or COR production, tvrR appears to be a novel virulence factor required for a previously unexplored process that is necessary for pathogenesis. PMID- 16267305 TI - Cell division defects in Escherichia coli deficient in the multidrug efflux transporter AcrEF-TolC. AB - The Escherichia coli chromosome contains several operons encoding confirmed and predicted multidrug transporters. Among these transporters only the inactivation of components of the AcrAB-TolC complex leads to substantial changes in susceptibility to multiple drugs. This observation prompted a conclusion that other transporters are silent or expressed at levels insufficient to contribute to multidrug resistance phenotype. We found that increased expression of AcrA, the periplasmic membrane fusion protein, is toxic only in cells lacking the multidrug efflux transporter AcrEF. AcrEF-deficient cells with increased expression of AcrA have a severe cell division defect that results in cell filamentation (>50 microm). Similar defects were obtained in cells lacking the outer membrane channel TolC, which acts with AcrEF, suggesting that cell filamentation is caused by the loss of AcrEF function. Green fluorescent protein AcrA fusion studies showed that in normal and filamentous cells AcrA is associated with membranes in a confined manner and that this localization is not affected by the lack of AcrEF. Similarly, the structure and composition of membranes were normal in filamentous cells. Fluorescence microscopy showed that the filamentous AcrEF-deficient E. coli cells are defective in chromosome condensation and segregation. Our results suggest that the E. coli AcrEF transporter is expressed under standard laboratory conditions and plays an important role in the normal maintenance of cell division. PMID- 16267307 TI - Polar localization of a soluble methyl-accepting protein of Pseudomonas aeruginosa. AB - A soluble methyl-accepting chemotaxis protein (MCP) of Pseudomonas aeruginosa, McpS, showed polar localization by immunofluorescence microscopy. Overexpression of McpS resulted in a dominant-negative effect on chemotaxis and caused a loss of polar clustering of the general MCP population. The polar localization of a soluble MCP defines a third, and unexpected, paradigm for cellular MCP localization. PMID- 16267306 TI - Catabolite repression and activation in Bacillus subtilis: dependency on CcpA, HPr, and HprK. AB - Previous studies have suggested that the transcription factor CcpA, as well as the coeffectors HPr and Crh, both phosphorylated by the HprK kinase/phosphorylase, are primary mediators of catabolite repression and catabolite activation in Bacillus subtilis. We here report whole transcriptome analyses that characterize glucose-dependent gene expression in wild-type cells and in isogenic mutants lacking CcpA, HprK, or the HprK phosphorylatable serine in HPr. Binding site identification revealed which genes are likely to be primarily or secondarily regulated by CcpA. Most genes subject to CcpA-dependent regulation are regulated fully by HprK and partially by serine-phosphorylated HPr [HPr(Ser-P)]. A positive linear correlation was noted between the dependencies of catabolite-repressible gene expression on CcpA and HprK, but no such relationship was observed for catabolite-activated genes, suggesting that large numbers of the latter genes are not regulated by the CcpA-HPr(Ser-P) complex. Many genes that mediate nitrogen or phosphorus metabolism as well as those that function in stress responses proved to be subject to CcpA-dependent glucose control. While nitrogen-metabolic genes may be subject to either glucose repression or activation, depending on the gene, almost all glucose-responsive phosphorus metabolic genes exhibit activation while almost all glucose-responsive stress genes show repression. These responses are discussed from physiological standpoints. These studies expand our appreciation of CcpA-mediated catabolite control and provide insight into potential interregulon control mechanisms in gram-positive bacteria. PMID- 16267308 TI - Alternate sigma factor RpoS is required for the in vivo-specific repression of Borrelia burgdorferi plasmid lp54-borne ospA and lp6.6 genes. AB - While numerous positively regulated loci have been characterized during the enzootic cycle of Borrelia burgdorferi, very little is known about the mechanism(s) involved in the repression of borrelial loci either during tick feeding or within the mammalian host. Here, we report that the alternative sigma factor RpoS is required for the in vivo-specific repression of at least two RpoD dependent B. burgdorferi loci, ospA and lp6.6. The downregulation of ospA and Ip6.6 appears to require either a repressor molecule whose expression is RpoS dependent or an accessory factor which enables RpoS to directly interact with the ospA and Ip6.6 promoter elements, thereby blocking transcription by RpoD. The central role for RpoS during the earliest stages of host adaptation suggests that tick feeding imparts signals to spirochetes that trigger the RpoS-dependent repression, as well as expression, of in vivo-specific virulence factors critical for the tick-to-mammalian host transition. PMID- 16267309 TI - The Chlamydia pneumoniae type III secretion-related lcrH gene clusters are developmentally expressed operons. AB - Two chlamydial homologues of the Yersinia lcrH chaperone for type III secretion system structural components are present within separate gene clusters. Quantitative transcriptional analyses demonstrated that each cluster is differentially regulated and expressed as an operon using major sigma factor elements, suggesting the presence of more elaborate developmental regulation mechanisms in chlamydiae. PMID- 16267310 TI - Actin-binding proteins from Burkholderia mallei and Burkholderia thailandensis can functionally compensate for the actin-based motility defect of a Burkholderia pseudomallei bimA mutant. AB - Recently we identified a bacterial factor (BimA) required for actin-based motility of Burkholderia pseudomallei. Here we report that Burkholderia mallei and Burkholderia thailandensis are capable of actin-based motility in J774.2 cells and that BimA homologs of these bacteria can restore the actin-based motility defect of a B. pseudomallei bimA mutant. While the BimA homologs differ in their amino-terminal sequence, they interact directly with actin in vitro and vary in their ability to bind Arp3. PMID- 16267311 TI - Streptococcus mutans murein hydrolase. AB - Allelic replacement of the C terminus of a Streptococcus mutans surface protein affects murein hydrolase activity. The targeted open reading frame encodes a 67 kDa protein (SmaA) with an N-terminal signal sequence and cleavage site, three 46 amino-acid (aa) direct repeats, and two 88-aa direct repeats. The identical autolytic profile was obtained using a sortase mutant (SrtA(-)). PMID- 16267312 TI - Tryptophan catabolism: identification and characterization of a new degradative pathway. AB - A new tryptophan catabolic pathway is characterized from Burkholderia cepacia J2315. In this pathway, tryptophan is converted to 2-amino-3-carboxymuconate semialdehyde, which is enzymatically degraded to pyruvate and acetate via the intermediates 2-aminomuconate and 4-oxalocrotonate. This pathway differs from the proposed mammalian pathway which converts 2-aminomuconate to 2-ketoadipate and, ultimately, glutaryl-coenzyme A. PMID- 16267313 TI - CadC activates pH-dependent expression of the Vibrio vulnificus cadBA operon at a distance through direct binding to an upstream region. AB - The Vibrio vulnificus cadBA genes were transcribed as a transcriptional operon by a single promoter, P(cadBA), which was activated by CadC in a pH-dependent manner. A direct interaction between CadC and the P(cadBA) DNA was demonstrated, and a CadC binding site centered at -233.5 was mapped by deletion analyses of P(cadBA) and confirmed by a DNase I protection assay. PMID- 16267315 TI - Images in clinical medicine. Actinomyces hepatic abscess with cutaneous fistula. PMID- 16267316 TI - Betting on a malaria vaccine. PMID- 16267314 TI - Identification of a novel essential two-component signal transduction system, YhcSR, in Staphylococcus aureus. AB - Two-component signal transduction systems play an important role in the ability of bacteria to adapt to various environments by sensing changes in their habitat and by altering gene expression. In this study, we report a novel two-component system, YhcSR, in Staphylococcus aureus which is required for bacterial growth in vitro. We found that the down-regulation of yhcSR expression by induced yhcS antisense RNA can inhibit and terminate bacterial growth. Moreover, without complementary yhcS or yhcR, no viable yhcS or yhcR gene replacement mutant was recoverable. Collectively, these results demonstrated that the YhcSR regulatory system is indispensable for S. aureus growth in culture. Moreover, induced yhcS antisense RNA selectively increased bacterial susceptibility to phosphomycin. These data suggest that YhcSR probably modulates the expression of genes critical for bacterial survival and may be a potential target for the development of novel antibacterial agents. PMID- 16267317 TI - Beyond Red Lake--the persistent crisis in American Indian health care. PMID- 16267318 TI - Paving the way--providing opportunities for Native American students. PMID- 16267319 TI - The complexity of microbial diversity in bacterial vaginosis. PMID- 16267320 TI - Prognostic significance of dyspnea in patients referred for cardiac stress testing. AB - BACKGROUND: Although dyspnea is a common symptom, there has been only limited investigation of its prognostic significance among patients referred for cardiac evaluation. METHODS: We studied 17,991 patients undergoing myocardial-perfusion single-photon-emission computed tomography during stress and at rest. Patients were divided into five categories on the basis of symptoms at presentation (none, nonanginal chest pain, atypical angina, typical angina, and dyspnea). Multivariable analysis was used to assess the incremental prognostic value of symptom categories in predicting the risk of death from cardiac causes and from any cause. In addition, the prognosis associated with various symptoms at presentation was compared in subgroups selected on the basis of propensity analysis. RESULTS: After a mean (+/-SD) follow-up of 2.7+/-1.7 years, the rate of death from cardiac causes and from any cause was significantly higher among patients with dyspnea (both those previously known to have coronary artery disease and those with no known history of coronary artery disease) than among patients with other or no symptoms at presentation. Among patients with no known history of coronary artery disease, those with dyspnea had four times the risk of sudden death from cardiac causes of asymptomatic patients and more than twice the risk of patients with typical angina. Dyspnea was associated with a significant increase in the risk of death among each clinically relevant subgroup and remained an independent predictor of the risk of death from cardiac causes (P<0.001) and from any cause (P<0.001) after adjustment for other significant factors by multivariable and propensity analysis. CONCLUSIONS: In a large series of patients, self-reported dyspnea identified a subgroup of otherwise asymptomatic patients at increased risk for death from cardiac causes and from any cause. Our results suggest that an assessment of dyspnea should be incorporated into the clinical evaluation of patients referred for cardiac stress testing. PMID- 16267321 TI - Molecular identification of bacteria associated with bacterial vaginosis. AB - BACKGROUND: Bacterial vaginosis affects millions of women and is associated with several serious health conditions. The cause of bacterial vaginosis remains poorly understood despite numerous studies based on cultures. Bacteria in microbial communities can be identified without cultivation by characterizing their ribosomal DNA (rDNA) sequences. METHODS: We identified bacteria in samples of vaginal fluid with a combination of broad-range polymerase-chain-reaction (PCR) amplification of 16S rDNA with clone analysis, bacterium-specific PCR assay of 16S rDNA, and fluorescence in situ hybridization (FISH) performed directly on vaginal fluid from 27 subjects with bacterial vaginosis and 46 without the condition. Twenty-one subjects were studied with the use of broad-range PCR of 16S rDNA, and 73 subjects were studied with the use of bacterium-specific PCR. RESULTS: Women without bacterial vaginosis had 1 to 6 vaginal bacterial species (phylotypes) in each sample (mean, 3.3), as detected by broad-range PCR of 16S rDNA, and lactobacillus species were the predominant bacteria noted (83 to 100 percent of clones). Women with bacterial vaginosis had greater bacterial diversity (P<0.001), with 9 to 17 phylotypes (mean, 12.6) detected per sample and newly recognized species present in 32 to 89 percent of clones per sample library (mean, 58 percent). Thirty-five unique bacterial species were detected in the women with bacterial vaginosis, including several species with no close cultivated relatives. Bacterium-specific PCR assays showed that several bacteria that had not been previously described were highly prevalent in subjects with bacterial vaginosis but rare in healthy controls. FISH confirmed that newly recognized bacteria detected by PCR corresponded to specific bacterial morphotypes visible in vaginal fluid. CONCLUSIONS: Women with bacterial vaginosis have complex vaginal infections with many newly recognized species, including three bacteria in the Clostridiales order that were highly specific for bacterial vaginosis. PMID- 16267322 TI - Natalizumab induction and maintenance therapy for Crohn's disease. AB - BACKGROUND: Natalizumab, a humanized monoclonal antibody against alpha4 integrin, inhibits leukocyte adhesion and migration into inflamed tissue. METHODS: We conducted two controlled trials to evaluate natalizumab as induction and maintenance therapy in patients with active Crohn's disease. In the first trial, 905 patients were randomly assigned to receive 300 mg of natalizumab or placebo at weeks 0, 4, and 8. The primary outcome was response, defined by a decrease in the Crohn's Disease Activity Index (CDAI) score of at least 70 points, at week 10. In the second trial, 339 patients who had a response to natalizumab in the first trial were randomly reassigned to receive 300 mg of natalizumab or placebo every four weeks through week 56. The primary outcome was a sustained response through week 36. A secondary outcome in both trials was disease remission (a CDAI score of less than 150). RESULTS: In the first trial, the natalizumab and placebo groups had similar rates of response (56 percent and 49 percent, respectively; P=0.05) and remission (37 percent and 30 percent, respectively; P=0.12) at 10 weeks. Continuing natalizumab in the second trial resulted in higher rates of sustained response (61 percent vs. 28 percent, P<0.001) and remission (44 percent vs. 26 percent, P=0.003) through week 36 than did switching to placebo. Serious adverse events occurred in 7 percent of each group in the first trial and in 10 percent of the placebo group and 8 percent of the natalizumab group in the second trial. In an open-label extension study, a patient treated with natalizumab died from progressive multifocal leukoencephalopathy, associated with the JC virus, a human polyomavirus. CONCLUSIONS: Induction therapy with natalizumab for Crohn's disease resulted in small, nonsignificant improvements in response and remission rates. Patients who had a response had significantly increased rates of sustained response and remission if natalizumab was continued every four weeks. The benefit of natalizumab will need to be weighed against the risk of serious adverse events, including progressive multifocal leukoencephalopathy. (ClinicalTrials.gov numbers, NCT00032786 and NCT00032799.) PMID- 16267323 TI - Congenital glutamine deficiency with glutamine synthetase mutations. AB - Glutamine synthetase plays a major role in ammonia detoxification, interorgan nitrogen flux, acid-base homeostasis, and cell signaling. We report on two unrelated newborns who had congenital human glutamine synthetase deficiency with severe brain malformations resulting in multiorgan failure and neonatal death. Glutamine was largely absent from their serum, urine, and cerebrospinal fluid. Each infant had a homozygous mutation in the glutamine synthetase gene (R324C and R341C). Studies that used immortalized lymphocytes expressing R324C glutamine synthetase (R324C-GS) and COS7 cells expressing R341C-GS suggest that these mutations are associated with reduced glutamine synthetase activity. PMID- 16267324 TI - Clinical practice. Allergic rhinitis. PMID- 16267325 TI - Bacterial infections in drug users. PMID- 16267326 TI - Images in clinical medicine. Traumatic cranial nerve palsy. PMID- 16267327 TI - Clinical problem-solving. A perfect storm. PMID- 16267328 TI - Dyspnea and risk in suspected coronary disease. PMID- 16267329 TI - Selective adhesion-molecule therapy and inflammatory bowel disease--a tale of Janus? PMID- 16267330 TI - Metabolic disorders in the center of genetic medicine. PMID- 16267331 TI - Echinacea angustifolia in rhinovirus infections. PMID- 16267332 TI - Adherence to medication. PMID- 16267333 TI - Postmenopausal osteoporosis. PMID- 16267334 TI - Screening for osteoporosis. PMID- 16267335 TI - Cabergoline-related severe restrictive mitral regurgitation. PMID- 16267336 TI - Statistical methods in the journal. PMID- 16267337 TI - Dynamic gene expression changes precede dioxin-induced liver pathogenesis in medaka fish. AB - A major challenge for environmental genomics is linking gene expression to cellular toxicity and morphological alteration. Herein, we address complexities related to hepatic gene expression responses after a single injection of the aryl hydrocarbon receptor (AHR) agonist 2,3,7,8-tetrachlorodibenzo-p-dioxin (dioxin) and illustrate an initial stress response followed by cytologic and adaptive changes in the teleost fish medaka. Using a custom 175-gene array, we find that overall hepatic gene expression and histological changes are strongly dependent on dose and time. The most pronounced dioxin-induced gene expression changes occurred early and preceded morphologic alteration in the liver. Following a systematic search for putative Ah response elements (AHREs) (5'-CACGCA-3') within 2000 bp upstream of the predicted transcriptional start site, the majority (87%) of genes screened in this study did not contain an AHRE, suggesting that gene expression was not solely dependent on AHRE-mediated transcription. Moreover, in the highest dosage, we observed gene expression changes associated with adaptation that persisted for almost two weeks, including induction of a gene putatively identified as ependymin that may function in hepatic injury repair. These data suggest that the cellular response to dioxin involves both AHRE- and non-AHRE-mediated transcription, and that coupling gene expression profiling with analysis of morphologic pathogenesis is essential for establishing temporal relationships between transcriptional changes, toxicity, and adaptation to hepatic injury. PMID- 16267338 TI - A Weight-of-evidence analysis of the cancer dose-response characteristics of 2,3,7,8-tetrachlorodibenzodioxin (TCDD). AB - Cancer risk assessment for TCDD and other compounds must focus on the cancer dose response relationship and corresponding potency for the range of human doses before it can have relevance to the human exposure environment. Major differences of opinion exist over whether the dose-response curve for TCDD and other dioxin congeners is non-linear (incorporating a threshold dose region below which tumors are unlikely to be elicited) or linear (implying that any exposure has a statistical likelihood of causing cancer). The World Health Organization and others strongly support a non-linear dose-response relationship for TCDD and cancer, whereas USEPA characterizes the dose-response function as linear. This review critically summarizes the available information on TCDD dose-response relationship for cancer utilizing a weight-of-evidence approach. This assessment concludes that the available data support a non-linear dose-response relationship as being most likely and appropriate for human cancer risk assessment, i.e., the evidence suggests that a biological threshold exists in the dose-response. While proof of a threshold is not absolute, and never can be, the level of certainty for TCDD is substantial because of the concordance of many lines of evidence and the consistency of repeated observations pointing to non-linearity. PMID- 16267339 TI - Drug bust. PMID- 16267340 TI - Developing a research agenda in biogerontology: basic mechanisms. AB - The National Institute on Aging (NIA) began operation in 1975, splitting off from the National Institute of Child Health and Human Development. The first 10 years of NIA's existence were characterized by funding descriptive and discovery research, as the field by then had not come of age. With the isolation of long lived animal mutants and the application of the tools of molecular biology (including whole-genome sequencing) and transgenic technology to biogerontology research, the situation has changed dramatically since then, and aging-related research has become increasingly mechanistic and respectable. This transition has been aided by research initiatives implemented by NIA staff, and the goal of this article is to describe how NIA develops such research initiatives using research progress made in biogerontology over the past 20 years as the basis for the discussion. PMID- 16267341 TI - Keeping priorities: the role of working memory and selective attention in cognitive aging. AB - Cognitive aging is associated with impairments to working memory and top-down control in selective attention, two components of cognitive control associated with the frontal lobes. Recent findings indicate that working memory and selective attention may be interdependent, making a better understanding of their involvement in cognitive aging particularly challenging. A paper in a recent issue of Nature Neuroscience has provided evidence that a reduction in the ability to keep a clear distinction between information to be stored in working memory and information that should be ignored and subsequently suppressed is associated with poor working memory performance. These results are in line with previous evidence for a specific age-related impairment in the ability to separate irrelevant from relevant information and may be able to explain a range of age-related cognitive changes. PMID- 16267344 TI - The science of COPD. Opportunities for combination therapy. Proceedings of a symposium. September 17-19, 2004. Miami, Florida, USA. PMID- 16267342 TI - Swelling of phospholipids by monovalent salt. AB - Critical to biological processes such as membrane fusion and secretion, ion-lipid interactions at the membrane-water interface still raise many unanswered questions. Using reconstituted phosphatidylcholine membranes, we confirm here that multilamellar vesicles swell in salt solutions, a direct indication that salt modifies the interactions between neighboring membranes. By varying sample histories, and by comparing with data from ion carrier-containing bilayers, we eliminate the possibility that swelling is an equilibration artifact. Although both attractive and repulsive forces could be modified by salt, we show experimentally that swelling is driven primarily by weakening of the van der Waals attraction. To isolate the effect of salt on van der Waals interactions, we focus on high salt concentrations at which any possible electrostatic interactions are screened. By analysis of X-ray diffraction data, we show that salt does not alter membrane structure or bending rigidity, eliminating the possibility that repulsive fluctuation forces change with salt. By measuring changes in interbilayer separation with applied osmotic stress, we have determined, using the standard paradigm for bilayer interactions, that 1 M concentrations of KBr or KCl decrease the van der Waals strength by 50%. By weakening van der Waals attractions, salt increases energy barriers to membrane contact, possibly affecting cellular communication and biological signaling. PMID- 16267343 TI - The biotin-capture lipid affinity assay: a rapid method for determining lipid binding parameters for apolipoproteins. AB - The lipid affinity of plasma apolipoproteins is an important modulator of lipoprotein metabolism. Mutagenesis techniques have been widely used to modulate apolipoprotein lipid affinity for studying biological function, but the approach requires rapid and reliable lipid affinity assays to compare the mutants. Here, we describe a novel method that measures apolipoprotein binding to a standardized preparation of small unilamellar vesicles (SUVs) containing trace biotinylated and fluorescent phospholipids. After a 30 min incubation at various apolipoprotein concentrations, vesicle-bound protein is rapidly separated from free protein on columns of immobilized streptavidin in a 96-well microplate format. Vesicle-bound protein and lipid are eluted and measured in a fluorescence microplate reader for calculation of a dissociation constant and the maximum number of potential binding sites on the SUVs. Using human apolipoprotein A-I (apoA-I), apoA-IV, and mutants of each, we show that the assay generates binding constants that are comparable to other methods and is reproducible across time and apolipoprotein preparations. The assay is easy to perform and can measure triplicate binding parameters for up to 10 separate apolipoproteins in 3.5 h, consuming only 120 microg of apolipoprotein in total. The benefits and potential drawbacks of the assay are discussed. PMID- 16267345 TI - Clinical Year in Review III: Sleep-disordered breathing, interstitial lung disease, lung transplantation, and pediatric pulmonary disease. PMID- 16267347 TI - Chronic obstructive pulmonary disease outcome measurements: what's important? What's useful? AB - The severity of chronic obstructive pulmonary disease (COPD) and patients' response to therapy are difficult to assess. The traditional measure, spirometry, correlates poorly with important clinical features of the disease, such as survival and quality of life (QOL). Moreover, COPD has recently been recognized as a systemic disease, and its systemic manifestations, such as weight loss and muscle weakness, are only poorly related to lung function. Therefore, although lung function remains an important outcome, other outcomes must be included in any overall assessment of disease severity or response to interventions. Examples include refinements of spirometry, such as measurement of FEV6 and inspiratory capacity; functional outcomes, such as dyspnea indexes and exercise tests; and global-clinical outcomes, such as QOL questionnaires and assessment of frequency and severity of acute exacerbations. For scoring disease severity, making a prognosis, or determining the outcome of novel interventions, composite measures need to be developed that take into account as many aspects of COPD as practicable. PMID- 16267348 TI - Combination therapy for chronic obstructive pulmonary disease: clinical aspects. AB - Anticholinergics and beta-agonists reduce bronchoconstriction through different mechanisms, and there is a long history of combination therapy with short-acting agents in these classes for chronic obstructive pulmonary disease. Such combinations may allow lower doses and thereby improve safety. Oral theophylline has also been combined with short-acting bronchodilators for many years. Most studies, however, show only mild improvements in bronchodilation at the expense of increased adverse effects. Professional society guidelines recommend that as the symptoms of chronic obstructive pulmonary disease progress, the patient should receive regular treatment with one or more long-acting bronchodilators, and an inhaled corticosteroid if the patient has repeated exacerbations. The combination of a short-acting anticholinergic with a long-acting beta-agonist, or the combination of a long-acting anticholinergic with a short- or long-acting beta-agonist, has been shown in most studies to improve lung function versus monotherapy with the individual components. Systematic reviews have concluded that fluticasone and salmeterol, and budesonide and formoterol, are superior to placebo and lead to clinically meaningful improvements in lung function, exacerbation rate, and quality of life. Effects on survival are less clear. Some of the other issues to be resolved are the safety of combination therapy, its pharmacoeconomic impact, and the role of newer agents. PMID- 16267349 TI - Pharmacologic principles for combination therapy. AB - This article discusses the pharmacologic basis for understanding the therapeutic actions of drugs, particularly for their use in combinations. The focus is on principles underlying combination therapy in general, including examples from diseases other than chronic obstructive pulmonary disease (COPD). Pharmacodynamic aspects of drug action are covered, with an emphasis on recent advances in the understanding of drug-receptor interactions and of drug agonism. Pharmacokinetics and drug-induced adaptive changes in receptors and cell signaling pathways are summarized, emphasizing their importance for potential combination therapies aimed at prolonging drug action. An organizational framework for three different approaches to combination therapy is then proposed; the molecular rationales for each approach are described together with classic examples from other diseases, and then their application to combination therapy in COPD is discussed. Finally, terminology for the independent and interactive effects of drug combinations is discussed, and approaches to the quantitative analysis and visual display of the effects of drug combinations are introduced. The basic principles reviewed here provide the pharmacologic foundation for subsequent articles in this issue that address the combinations in current use for COPD, and they point to novel strategies for potential future approaches to combination therapy in COPD. PMID- 16267346 TI - Pathogenesis of chronic obstructive pulmonary disease. AB - The current paradigm for the pathogenesis of chronic obstructive pulmonary disease is that chronic airflow limitation results from an abnormal inflammatory response to inhaled particles and gases in the lung. Airspace inflammation appears to be different in susceptible smokers and involves a predominance of CD8+ T lymphocytes, neutrophils, and macrophages. Studies have characterized inflammation in the peripheral airspaces in different stages of disease severity. Two other processes have received considerable research attention. The first is a protease-antiprotease imbalance, which has been linked to the pathogenesis of emphysema. However, the hypothesis of an increased protease burden associated with functional inhibition of antiproteases has been difficult to prove and is now considered an oversimplification. The second process, oxidative stress, has a role in many of the pathogenic processes of chronic obstructive pulmonary disease and may be one mechanism that enhances the inflammatory response. In addition, it has been proposed that the development of emphysema may involve alveolar cell loss through apoptosis. This mechanism may involve the vascular endothelial growth factor pathway and oxidative stress. PMID- 16267351 TI - Molecular mechanisms of beta2-adrenergic receptor function and regulation. AB - It is now clear that the beta2-adrenergic receptor continuously oscillates between various conformations in the basal state, and that agonists act to stabilize one or more conformations. It is conceivable that synthetic agonists might be engineered to preferentially confine the receptor to certain conformations deemed clinically important while having a less stabilizing effect on unwanted conformations. In addition, studies of genetically engineered mice have revealed previously unrecognized cross-talk between the beta2-receptor and phospholipase C, such that removal of the primary dilating pathway results in downregulation of constrictive pathways and overactivity of the dilating pathway increases the contractile response. These results indicate a dynamic interaction between beta2-receptor activity and Gq-coupled receptors that constrict the airway. Potentially, then, during chronic beta-agonist therapy, expression of phospholipase C is increased, the functions of Gq-coupled constrictive receptors are enhanced, and there may be an increased tendency for clinical decompensation due to asthma and chronic obstructive pulmonary disease triggers. Antagonists to these receptors might be able to act synergistically with chronic beta-agonists to block the effect of phospholipase C. Alternatively, perhaps novel phospholipase C antagonists would provide the most efficacious approach to blocking the physiologic sequelae of cross-talk between the beta2-receptor and phospholipase C. PMID- 16267352 TI - Cholinergic pathways in the lungs and anticholinergic therapy for chronic obstructive pulmonary disease. AB - Abundant data from animal models and humans support the hypothesis that changes at the level of parasympathetic neuronal control of airway smooth muscle result in increased bronchoconstriction in response to vagal stimulation, leading to airway hyperresponsiveness. Neuronal inhibitory M2 muscarinic acetylcholine receptors on parasympathetic nerves are responsible for limiting acetylcholine release from these nerves. In humans with asthma, and after pulmonary inflammatory events in experimental animals, these receptors are dysfunctional, which results in airway hyperresponsiveness. Although it is unknown what mechanisms underlie airway hyperresponsiveness in chronic obstructive pulmonary disease, loss of parasympathetic control of airway smooth muscle is thought to be a contributing mechanism. As such, anticholinergic therapy is used extensively and with a high degree of success in the treatment of this condition. The future for inhaled anticholinergic compounds for the treatment of chronic obstructive pulmonary disease appears to rest in their combination with other agents, such as beta2 agonists and phosphodiesterase-4 inhibitors. Nonselective anticholinergic agents might be the best choice, because M2 muscarinic receptors on airway smooth muscle inhibit the generation and accumulation of cyclic adenosine monophosphate. Adequate concurrent blockade of M3 muscarinic receptors would be expected to counteract the enhanced acetylcholine release that would result from blockade of neuronal inhibitory M2 muscarinic receptors. PMID- 16267353 TI - Beta2-agonist and anticholinergic drugs in the treatment of lung disease. AB - The lungs are innervated by both the sympathetic and parasympathetic nervous systems, which entails the activation of adrenergic and muscarinic receptors, respectively. Both the adrenergic and muscarinic receptors are G-protein-coupled receptors, and they share many similar signal transduction molecules. These receptors are widely expressed in the lung and the specific receptor expression can vary among the species. The location and the subtype of receptor expressed are important in the regulation of normal airway function. Acetylcholine released from the parasympathetic fibers activates the M3 muscarinic receptors located on the airway smooth muscle, causing bronchoconstriction. To counter this activity, M2 muscarinic receptors located on the parasympathetic nerves inhibit release of acetylcholine. Beta2-adrenergic receptors are expressed on the airway smooth muscle where activation causes bronchodilation. Adrenergic receptors are also on the autonomic nerves where they can modulate neurotransmitter release. The crosstalk between these G-protein-coupled receptors and downstream pathways ensures normal airway function. The prejunctional and postjunctional muscarinic and adrenergic receptors control autonomic tone and any imbalance or selective blockade of the receptors can compromise the system and cause the airways to become hyperreactive. The location, function, and crosstalk of the adrenergic and muscarinic receptors must be considered in the design, development, and use of drugs to combat airway diseases. PMID- 16267356 TI - Corticosteroids: potential beta2-agonist and anticholinergic interactions in chronic obstructive pulmonary disease. AB - Corticosteroids are often used in combination with beta2-agonist and anticholinergic bronchodilators in the treatment of chronic obstructive pulmonary disease (COPD). Corticosteroids activate the beta2-receptor gene, increasing receptor number and decreasing desensitization. Long-acting beta2-agonists prime the glucocorticoid receptor and enhance nuclear translocation via activation of CCAAT enhancer binding protein-alpha. Corticosteroids can also increase prejunctional auto-inhibitory M2-receptor gene expression in airway smooth muscle. There is evidence of a synergistic inhibition of cytokine and chemokine release from alveolar macrophages, epithelial cells, and mucosal glands and enhanced respiratory cytoprotection against viral and bacterial infection when a corticosteroid is combined with salmeterol. In airway smooth muscle, corticosteroids inhibit the contractile effects of acetylcholine, whereas M2 receptor antagonism increases the relaxant activity of isoproterenol. Complementary interactions between corticosteroids and long-acting beta2-agonists and between corticosteroids and anticholinergic bronchodilators may be important if these drugs are combined in the treatment of COPD. PMID- 16267355 TI - Glucocorticoid pathways in chronic obstructive pulmonary disease therapy. AB - Lung function measures in patients with chronic obstructive pulmonary disease remain insensitive to corticosteroid actions, in contrast to the clinical improvements observed in most patients with asthma. By uncovering the reason for this paradox, physicians should be able to implement treatment regimens that restore corticosteroid sensitivity. Corticosteroids exert their effects by binding to a cytoplasmic glucocorticoid receptor, which is subjected to post translational modification by phosphorylation. Receptor phosphorylation may influence hormone binding and nuclear translocation, as well as alter other glucocorticoid receptor interactions, its protein half-life, and downregulation processes. This suggests that a "phosphorylation code" may exist for glucocorticoid receptor function. Oxidative stress due to cigarette smoke may also be a mechanism for the corticosteroid resistance observed in chronic obstructive pulmonary disease, as it enhances proinflammatory transcription. Reduced glucocorticoid nuclear translocation along with attenuated histone deacetylase activity may be partially responsible for this effect. Therapies targeting these aspects of the glucocorticoid receptor activation pathway may reverse steroid resistance in patients with chronic obstructive pulmonary disease. PMID- 16267357 TI - Phosphodiesterase-4: selective and dual-specificity inhibitors for the therapy of chronic obstructive pulmonary disease. AB - Phosphodiesterase-4 isoenzymes have absolute specificity for cyclic adenosine 3',5'-monophosphate and are considered potential therapeutic targets for the treatment of chronic inflammatory disorders, such as chronic obstructive pulmonary disease, with small-molecule inhibitors. Several selective phosphodiesterase-4 inhibitors are in clinical trials of chronic obstructive pulmonary disease, including cilomilast and roflumilast. Despite some encouraging data from phase III clinical trials, the current generation of phosphodiesterase 4 inhibitors is hampered by a low therapeutic ratio. Indeed, a major obstacle is their propensity to evoke non-steroid-like side effects, of which nausea, diarrhea, abdominal pain, vomiting, and dyspepsia are the most common. In addition, a particularly worrying potential toxicity of phosphodiesterase-4 inhibitors, also shared by phosphodiesterase-3 inhibitors and other vasodilators, is arteritis/periarteritis. One potential means of improving the therapeutic ratio and safety of phosphodiesterase-4 inhibitors may lie in the development of compounds that have broader phosphodiesterase specificity. Of the 11 phosphodiesterase families that have been unequivocally identified, dual specificity compounds that inhibit phosphodiesterase-4 and phosphodiesterase-1, phosphodiesterase-3, or phosphodiesterase-7 may offer the best opportunities to enhance clinical efficacy. PMID- 16267358 TI - Theophylline in chronic obstructive pulmonary disease: new horizons. AB - Although theophylline has side effects when used in bronchodilator doses, increasing evidence shows that it has significant antiinflammatory effects in chronic obstructive pulmonary disease at lower plasma concentrations. These antiinflammatory effects are unlikely to be accounted for by phosphodiesterase inhibition or adenosine receptor antagonism, which require higher concentrations. There is now evidence that theophylline at low therapeutic concentrations is an activator of histone deacetylases and that this activation enhances the antiinflammatory effect of corticosteroids. There appears to be a marked reduction in histone deacetylase-2 in macrophages and peripheral lung of patients with chronic obstructive pulmonary disease, which accounts for amplified inflammation and steroid resistance. Theophylline has been shown to restore steroid sensitivity in vitro. The effect of theophylline on histone deacetylase activity appears to be enhanced by oxidative stress. The mechanism whereby theophylline activates histone deacetylase is not yet known, but it does not involve other known actions of theophylline that account for its side effects. Better understanding of the molecular basis for the action of theophylline might lead to the development of novel drugs. PMID- 16267360 TI - Innate immune responses and chronic obstructive pulmonary disease: "Terminator" or "Terminator 2"? AB - Innate immune responses appear to be partially responsible for maintaining inflammation and tissue destruction in chronic obstructive pulmonary disease. In the early stages of the disease in smokers, the airways are bombarded with large quantities of particulate material, and activation of phagocytic cells results in the release of many of the mediators believed to remodel the airways. Ironically, failure of the innate immune defense system, either by inherited deficiency or as a result of chronic smoke inhalation, is likely to result in increased susceptibility to infectious disease and exacerbations of chronic obstructive pulmonary disease. It is well known that deficiencies in the production of collectins, pentraxins, and complement can lead to increased infections, and several studies indicate that deficiency in one or another innate defense component is associated with increased exacerbations. Corticosteroids reduce exacerbations in part because of their ability to boost the production of innate host-defense molecules. Therapeutic approaches that stimulate the generation of antimicrobial molecules in the lungs might be able to reduce disease exacerbations. PMID- 16267361 TI - The role of airway smooth muscle in the pathogenesis of airway wall remodeling in chronic obstructive pulmonary disease. AB - Airway wall remodeling processes are present in the small airways of patients with chronic obstructive pulmonary disease, consisting of tissue repair and epithelial metaplasia that contribute to airway wall thickening and airflow obstruction. With increasing disease severity, there is also increased mucous metaplasia and submucosal gland hypertrophy, peribronchial fibrosis, and an increase in airway smooth muscle mass. Apart from its contractile properties, airway smooth muscle produces inflammatory cytokines, proteases, and growth factors, which may contribute to the remodeling process and induce phenotypic changes of the muscle. Airflow limitation responds minimally to beta-agonists and corticosteroid therapy, unlike asthma, perhaps because of alterations in beta receptor or glucocorticoid receptor numbers, alterations in receptor signaling, or the constrictive limitation imposed by peribronchial fibrosis. Better response is observed with the combination of inhaled long-acting beta-agonists and corticosteroids. This could result from effects at the level of airway smooth muscle. Airway wall remodeling may involve the release of growth factors from inflammatory or resident cells. The influence of smoking cessation or of current therapies on airway wall remodeling is unknown. Specific therapies for airway wall remodeling may be necessary, together with noninvasive methods of imaging small airway wall remodeling to assess responses. PMID- 16267363 TI - Mechanisms and experimental models of chronic obstructive pulmonary disease exacerbations. AB - Exacerbations of chronic obstructive pulmonary disease (COPD) are a major cause of morbidity, mortality, and rising health care costs. In addition, they are associated with an accelerated loss of lung function and thus have a direct effect on disease progression. There are few studies examining the cellular and molecular mechanisms of COPD exacerbations. Exacerbations are linked to increased airway inflammation and oxidative stress, but many questions remain unanswered regarding the key inflammatory cells and mediators. Current therapies for COPD exacerbations are of limited effectiveness, and a better understanding of the inflammatory events at exacerbation is required to devise new therapeutic agents. The development of experimental models of exacerbation-for example, the use of experimental rhinovirus infection in humans with COPD-would greatly facilitate studies of exacerbations. PMID- 16267364 TI - Systemic effects of chronic obstructive pulmonary disease. AB - Chronic obstructive pulmonary disease (COPD) affects various structural and functional domains in the lungs. It also has significant extrapulmonary effects, the so-called systemic effects of COPD. Weight loss, nutritional abnormalities, and skeletal muscle dysfunction are well-recognized systemic effects of COPD. Other less well-known but potentially important systemic effects include an increased risk of cardiovascular disease and several neurologic and skeletal defects. The mechanisms underlying these systemic effects are unclear, but they are probably interrelated and multifactorial, including inactivity, systemic inflammation, tissue hypoxia and oxidative stress among others. These systemic effects add to the respiratory morbidity produced by the underlying pulmonary disease and should be considered in the clinical assessment as well as the treatment of affected patients. PMID- 16267362 TI - The role of vagal afferent nerves in chronic obstructive pulmonary disease. AB - Circumstantial evidence supports the hypothesis that the vagal nervous system is dysregulated in chronic obstructive pulmonary disease. This dysregulation can lead to an increased sensitivity of the cough reflex such that the coughing becomes, at times, "nonproductive" or inappropriate. Vagal dysregulation can also lead to an increase in the activity of the parasympathetic reflex control of the airways, which contributes to greater mucus secretion and bronchial smooth muscle contraction. Indirect evidence indicates that lung disease is accompanied by substantive changes to the entire reflex pathways, including enhanced activity of the primary afferent nerves, increases in synaptic efficacy at secondary nerves in the central nervous system, and changes in the autonomic nerve pathways. Drugs aimed at normalizing neuronal activity may, therefore, be beneficial in chronic obstructive pulmonary disease. PMID- 16267368 TI - Challenges and opportunities for combination therapy in chronic obstructive pulmonary disease. AB - Advances in the understanding of chronic obstructive pulmonary disease have presented a number of novel therapeutic opportunities. More extensive use of drug combinations is likely, but the development of these therapies presents a number of challenges. In clinical trials, a combination must be tested not only against placebo but also against each of its components, and the false-positive error rate increases rapidly with multiple comparisons. Thus, more groups of subjects must be studied, and more individuals within each group must be studied, in order to ensure statistical significance. Another challenge is that the relatively slow progression of chronic obstructive pulmonary disease and the lack of specificity of commonly used outcome variables complicate the evaluation of all therapies, including combinations. In analogy to genomics and proteomics (evaluation of the pattern of expression of many things simultaneously), it may be more useful to adopt an approach that is here dubbed "clinicomics": consideration of multiple features of chronic obstructive pulmonary disease that are evaluated routinely, for example, with a well-done history and physical examination. The use of a truly multidimensional outcome parameter promises an entirely novel paradigm for the assessment of novel combinations of therapies. PMID- 16267367 TI - Inflammation-activated protein kinases as targets for drug development. AB - Given the prevalence and debilitating nature of chronic inflammatory diseases, there is a never-ending quest to identify novel targets for the rational development of antiinflammatory drugs. The major signaling pathway that controls inflammation-associated gene expression is the one which leads to activation of transcription factor nuclear factor-kappaB. Therefore, inhibitors of the kinase responsible for nuclear factor-kappaB activation, IkappaB kinase, are expected to have potent antiinflammatory activity. Indeed, our results with cell type specific inactivation of the beta-catalytic subunit of IkappaB kinase are by and large consistent with this assertion. In addition to IkappaB kinase and nuclear factor-kappaB, the expression of certain proinflammatory cytokines, such as interleukin-1 and tumor necrosis factor-alpha, is dependent on mitogen-activated protein kinases. Therefore, considerable attention has also been given to mitogen activated protein kinases as likely targets for the development of novel antiinflammatory therapeutics. Preliminary preclinical data suggest that inhibitors that target all these pathways exhibit antiinflammatory activity. This review focuses on the possible mechanisms through which such inhibitors may interfere with inflammation and some of the complications that may be associated with their use. PMID- 16267370 TI - A walking program for outpatients in psychiatric rehabilitation: pilot study. AB - The purposes of this quasi-experimental pilot study were to determine adherence to a 12-week group-based moderate-intensity walking program for sedentary adult outpatients with serious and persistent mental illness and to examine change from baseline to after the walking program in health status (mental and physical health, mood, and psychosocial functioning) and exercise motivation (exercise outcomes expectancies, exercise decisional balance). The 15 volunteers in this study were aged 21 to 65 years and enrolled in psychosocial rehabilitation; they participated in a 12-week walking program meeting three times per week for 1 hr, supplemented with four health information workshops delivered at the beginning of the study. Participants received individual exercise prescriptions determined by preprogram fitness testing and used heart rate monitors during walking sessions. Thirteen participants (87%) completed the study and attended 76% of the walking sessions. Overall, they walked at lower intensity than prescribed, with pulses within target heart rate ranges 35% of the time during Weeks 1 through 4, 26% of the time during Weeks 5 through 8, and 22% of the time during Weeks 9 through 12. However, mood improved (Profile of Mood States, t = -2.51, two-tailed, df = 12, p = .02), as did psychosocial functioning (Multnomah Community Ability Scale, two tailed, df = 12, t = 2.49, p = .02). The findings indicate a walking group may be feasible for rehabilitation programs. In addition to the known cardiovascular risk-reduction benefits of regular walking, walking may improve mood and psychosocial functioning in adults with serious and persistent mental illness. PMID- 16267371 TI - The effect of therapeutic back massage in hypertensive persons: a preliminary study. AB - Hypertension, one of the most pervasive disease processes in the United States, can lead to target organ damage. Although there is no one cause of primary hypertension, the theory of an unchecked long-term stress response continues to be a valid argument. Conversely, eliciting the relaxation response may alter the course of the unchecked stress response. Massage therapists have suggested that their therapy elicits the relaxation response and therefore can decrease blood pressure (BP) and hypertension. This preliminary study tested the effects of a regularly applied back massage on the BP of patients with clinically diagnosed hypertension. In this experimental, pretest-posttest study, a 10-min back massage was given to the experimental group (n = 8), three times a week for 10 sessions. The control group (n = 6) relaxed in the same environment for 10 min, three times a week for 10 sessions. Analysis of variance determined systolic BP changed significantly, F(1, 12) = 17.90, p = .001, between groups over time as did the diastolic BP, F(1, 12) = 8.34, p = .014. Effect size was 2.25 for systolic pressure and 1.56 for diastolic pressure (alpha of .05 and power at .80). This preliminary study suggests that regular massage may lower BP in hypertensive persons. PMID- 16267366 TI - Proteinases and oxidants as targets in the treatment of chronic obstructive pulmonary disease. AB - There is now compelling evidence that proteinases and oxidative stress play pathogenetic roles in the following pathologies in chronic obstructive pulmonary disease: airspace enlargement; chronic inflammation in the airways, lung interstitium, and alveolar space; and mucus hypersecretion in the large airways. Proteinases and oxidants may also contribute to remodeling processes in the small airways. In addition, data are emerging that show interactions between classes of proteinases and between proteinases and oxidants, which amplify lung inflammation and injury in chronic obstructive pulmonary disease. This review discusses the biologic roles of proteinases and oxidants, their roles in the pathogenesis of chronic obstructive pulmonary disease, and their potential as targets for therapy. PMID- 16267373 TI - Chronic inflammation and breast pathology: a theoretical model. AB - Breast cell pathology results from biochemical and molecular changes that culminate in the cell's loss of functional responsiveness. The epithelial cell compartment in the breast ductal system is the site of approximately 98% of malignant proliferations, and it is from within these cells that the first biochemical signal of change may be expressed as an inflammatory response. Inflammation may be represented by biomarkers of early pathologic changes in breast cells and be associated with risk for the development of breast cancer. A theoretical model of the inflammatory process is proposed showing predictive linkages among stimuli in the breast microenvironment and the development of breast pathology, in particular, breast cancer. This model fuels intervention concepts that may prevent malignant breast health outcomes. PMID- 16267372 TI - Differences between exclusive breastfeeders, formula-feeders, and controls: a study of stress, mood, and endocrine variables. AB - The purpose of this study was to examine relationships among lactational status, naturalistic stress, mood, and levels of serum cortisol and prolactin and plasma adrenocorticotropic hormone (ACTH). Eighty-four exclusively breastfeeding, 99 exclusively formula-feeding, and 33 nonpostpartum healthy control women were studied. The postpartum mothers were studied cross-sectionally once between 4 and 6 weeks after the birth. Stress was measured using the Perceived Stress Scale, the Tennessee Postpartum Stress Scale, and the Inventory of Small Life Events. Mood was measured using the Profile of Mood States. Serum prolactin, plasma ACTH, and serum cortisol levels were measured by commercial ELISA (enzyme-linked immunosorbent assay) kits. Results indicate that breastfeeding mothers had more positive moods, reported more positive events, and perceived less stress than formula-feeders. Reports of stressful life events were generally equivalent in the two groups. Serum prolactin was inversely related to stress and mood in formula-feeders. When breast and formula-feeders were compared to controls, they had higher serum cortisol, lower stress, and lower anxiety. Breastfeeders had lower perceived stress than controls. Breastfeeders had lower depression and anger and more positive life events reported than formula-feeders. However, there were few correlations among stress, mood, and the hormones in postpartum mothers, and those only in formula-feeders, whereas strong relationships were found between serum ACTH and a number of stress and mood variables in controls. Postpartum mothers reported a range of stress and negative moods at 4 to 6 weeks, and in formula-feeders, serum prolactin was related to some of the stress and mood variables. Breastfeeding appears to be somewhat protective of negative moods and stress. PMID- 16267374 TI - Physiological wet laboratory facilities in colleges of nursing. AB - The purpose of this study was to describe physiological wet laboratories as they exist within colleges of nursing with doctoral programs. Surveys were sent to the current deans and directors of all 96 nursing colleges/schools with doctoral programs as of January 2004, obtained from the American Association of Colleges of Nursing Web site. Only 26 (37%) of 71 responding schools operate their own wet laboratory, either singly or with another college. The most common analyses done are hormone levels, immune proteins, and cell counts, followed by catecholamines, chemistries, and bacterial cultures. Approximate annual costs of running wet laboratories range from 1,500 dollars to 320,000 dollars per school. Schools based in an academic health center were more likely to operate a wet laboratory ( chi2 = .003). There were no differences based on source of university funding (public vs. private) or Carnegie classification. PMID- 16267376 TI - Vascular responses to extractable fractions of Ilex paraguariensis in rats fed standard and high-cholesterol diets. AB - The authors investigated the vasorelaxant properties of the aqueous (Aq-EF) and acid n-butanolic (acn-BuOH) extractable fractions from Ilex paraguariensis leaves. Perfusion pressure was evaluated using isolated and perfused mesenteric arterial beds (MABs) from rats fed hypercholesterolemic and standard diets. Extract-induced vasorelaxation in the presence and absence of various inhibitors was examined following precontraction of the MABs with methoxamine (30 microM) solution. In hypercholesterolemic-diet rats, relaxation in intact MABs was significantly decreased with ac-n-BuOH-EF bolus (300, 600, 900 microg) in comparison to those in standard-diet rats. After the endothelium was stripped from the MABs, the vascular responses to ac-n-BuOH-EF and 900 microg bolus of Aq EF were significantly changed. Treatment of the MABs with an inhibitor of nitric oxide synthase, N(G)-nitro-L-arginine methylester hydrochloride (L-NAME, 10 mM), did not change either ac-n-BuOH-EF- or Aq-EF-induced vasodilation except for the 900 microg bolus of Aq-EF. The guanilate cyclase inhibitor methylene blue (100 microM) did not affect vasodilation for either fraction in the MABs from the hypercholesterolemic-diet rats. The chronic oral administration of I. paraguariensis extract in hypercholesterolemic-diet rats resulted in a significant reduction in serum levels of cholesterol and triglycerides. These results suggest that I. paraguariensis ac-n-BuOH-EF and Aq-EF induce vasodilation in standard-diet rats in a dose-dependent manner and that the hypercholesterolemic diet substantially reduced the effect of ac-n-BuOH-EF on precontracted MABs. PMID- 16267377 TI - Rapid changes in plant genomes. PMID- 16267375 TI - Effects of estrogen on platelet reactivity after transient forebrain ischemia in rats. AB - Estrogen's prothrombotic effects are of increasing concern, particularly in stroke risk and recovery. Using an ischemic rodent model, the authors sought to determine (a) if estrogen replacement increases postischemic platelet reactivity, (b) if changes in estrogen status alter intraplatelet endothelial nitric oxide synthase (eNOS) synthesis, and (c) if estrogen-mediated effects on platelets alter cerebral blood flow during reperfusion. Intact (I), ovariectomized (OVX), and OVX + 17 beta-estradiol (E50) rats were subjected to 30 min of forebrain ischemia and 60 min of reperfusion. Using the platelet activation marker P selectin, postischemic platelet reactivity was quantified by flow cytometry. In a separate cohort (I, OVX, E50), the authors quantified platelet eNOS by Western blot. Another cohort (OVX, E50) was subjected to ischemia/reperfusion, and cerebral blood flow was determined using the iodoantipyrine technique. Collagen stimulated platelet P-selectin expression was increased in the OVX rats at 60 min of reperfusion, and this effect was reversed by estrogen treatment. No differences in platelet eNOS expression were detected among groups. Cerebral blood flow at 60 min reperfusion was comparable between the OVX and the E50 rats. The authors conclude that during reper-fusion, estrogen deficiency increases postischemic platelet sensitivity to stimuli in estrogen-deficient rats. Estrogen treatment mitigates effects of estrogen loss on platelets, but this early effect is apparently not caused by intraplatelet eNOS depression. Neither estrogen deficiency nor estrogen treatment changes early postischemic regional brain blood flow. In this rodent global cerebral ischemic model, physiologic doses of estrogen are not deleterious to platelet reactivity and may initially reduce postischemic platelet reactivity. PMID- 16267378 TI - A toxic mutator and selection alternative to the non-Mendelian RNA cache hypothesis for hothead reversion. PMID- 16267379 TI - Differential control of Bmal1 circadian transcription by REV-ERB and ROR nuclear receptors. AB - Circadian rhythms result from feedback loops involving clock genes and their protein products. In mammals, 2 orphan nuclear receptors, REV-ERBalpha and RORalpha, play important roles in the transcription of the clock gene Bmal1. The authors now considerably extend these findings with the demonstration that all members of the REV-ERB (alpha and beta) and ROR (alpha, beta, and gamma) families repress and activate Bmal1 transcription, respectively. The authors further show that transcription of Bmal1 is the result of competition between REV-ERBs and RORs at their specific response elements (RORE). Moreover, they demonstrate that Reverb genes are similarly expressed in the thymus, skeletal muscle, and kidney, whereas Ror genes present distinct expression patterns. Thus, the results indicate that all members of the REV-ERB and ROR families are crucial components of the molecular circadian clock. Furthermore, their strikingly different patterns of expression in nervous and peripheral tissues provide important insights into functional differences between circadian clocks within the organism. PMID- 16267380 TI - Tissue-specific disruption of rhythmic expression of Dec1 and Dec2 in clock mutant mice. AB - DEC1 and DEC2-basic helix-loop-helix transcription factors-exhibit a circadian expression in the suprachiasmatic nucleus and other peripheral tissues and seem to play roles in regulating the mammalian circadian rhythm by suppressing the CLOCK/BMAL1-activated promoters of Per1, Dec1, and Dec2. The authors present data on the expression patterns of mRNA for Dec1, Dec2, Per2, Dbp, and Npas2 in various tissues of wild-type and homozygous Clock mutant mice (Clock/Clock). The Clock mutation resulted in extreme reduction of Dec1 expression in kidney, heart, and skeletal muscle but not in liver, whereas it strongly repressed Dec2 expression in liver, kidney, and heart, while Dec2 expression in skeletal muscle remained rhythmic. Per2 also showed the tissue-dependent disruption of the rhythmicity by Clock mutation, whereas rhythmic expression of Dbp in Clock mutant mice disappeared in all tissues examined. Npas2, a structurally and functionally related gene to Clock, showed significant levels of expression in the liver and kidney with a robust rhythmicity, which was also affected by Clock mutation. These marked changes in the Dec1 and Dec2 expression, as well as in the Per2, Dbp, and Npas2 expression in the periphery by Clock mutation, indicated that CLOCK plays a major role in the expression of these genes in most tissues. However, circadian expression of Dec1 in liver and kidney and that of Dec2 in skeletal muscle of Clock mutant mice suggested that CLOCK-independent circadian regulation operates in some tissues. PMID- 16267382 TI - Free-running rhythms of pineal circadian output. AB - Circadian rhythms are self-sustaining oscillations that free-run in constant conditions with a period close to 24 h. Overt circadian rhythms have been studied mostly using onset phase as the marker for the underlying pacemaker. Using in vivo online pineal microdialysis, the authors have performed detailed analysis of free-running profiles of rat pineal secretory products, including N acetylserotonin (NAS) and melatonin that have precisely defined onsets and offsets. When rats entrained in LD 12:12 were released into constant darkness (DD), both onset and offset phases of melatonin and NAS free-run. However, while onsets free-run with a period closer to a day (FRP(on) = 24-24.17 h) at the beginning, offset phases free-run with significantly larger FRPs (free-running periods) (FRP(off) = 24.24-24.42 h). This asymmetric free-running of onset and offset of NAS and melatonin in DD resulted in a 60- to 120-min increase of secretion duration of both NAS and melatonin. The rate of expansion of melatonin duration was 10 to 15 min per circadian cycle. The expansion of melatonin secretion duration ended for some within 4 days, while others were still expanding by the end of 10th day in DD. These results revealed that upon release into DD, the pacemaker's oscillation is initially driven by 2 forces, free running and decompression, before reaching a stable state of free running, and suggest that the circadian pacemaker may be an elastic structure that can decompress and compress under varying photic conditions. They also illustrate the importance of using both onset and offset of a given rhythm as phase markers, as compression/decompression, and transient disparity between FRP(on) and FRP(off) may be a common phenomenon of the circadian pacemaker. PMID- 16267383 TI - Reentrainment of the circadian pacemaker through three distinct stages. AB - Circadian rhythms are endogenously generated by a central pacemaker and are synchronized to the environmental LD cycle. The rhythms can be resynchronized, or reentrained, after a shift of the LD cycle, as in traveling across time zones. The authors have performed high-resolution mapping of the pacemaker to analyze the reentrainment process using rat pineal melatonin onset (MT(on)) and melatonin offset (MT(off)) rhythms as markers. Following LD (12:12) delays of 3, 6, and 12 h, MT(on) was phase locked immediately, whereas MT(off) shifted rapidly during the initial 1 through 3 cycles. In all animals, the MT(off) shifted beyond their expected phase positions in the new LD cycle, which resulted in a transient expansion of melatonin secretion duration for several cycles. It took MT(off) only 1, 2, or 3 cycles to complete most of the required phase shifts after 3, 6, or 12 h of the LD cycle delays, respectively. However, the final stabilization of phase relationships of both MT(on) and MT(off) required at least 6 cycles for rats experiencing a 3-h LD delay and much longer for the rest. These results reaffirmed the notion that both onset and offset phases of melatonin rhythms are important markers for the pacemaker and demonstrated that the reentrainment of the central pacemaker to a delay shift of the LD cycle is a 3-step process: an immediate phase lock of onset and a rapid delay shift of offset rhythms, overshoot of the offset, and, finally, a slow adjustment of both onset and offset phases. This study represents the 1st detailed analysis of the pacemaker behavior during reentrainment using melatonin and supports the notion that the eventual adaptation of the circadian pacemaker to a new time zone is a time-consuming process. PMID- 16267381 TI - c-Fos expression in the brains of behaviorally "split" hamsters in constant light: calling attention to a dorsolateral region of the suprachiasmatic nucleus and the medial division of the lateral habenula. AB - "Splitting" of circadian activity rhythms in Syrian hamsters maintained in constant light appears to be the consequence of a reorganized SCN, with left and right halves oscillating in antiphase; in split hamsters, high mRNA levels characteristic of day and night are simultaneously expressed on opposite sides of the paired SCN. To visualize the splitting phenomenon at a cellular level, immunohistochemical c-Fos protein expression in the SCN and brains of split hamsters was analyzed. One side of the split SCN exhibited relatively high c-Fos levels, in a pattern resembling that seen in normal, unsplit hamsters during subjective day in constant darkness; the opposite side was labeled only within a central-dorsolateral area of the caudal SCN, in a region that likely coincides with a photo-responsive, glutamate receptor antagonist-insensitive, pERK expressing cluster of cells previously identified by other laboratories. Outside the SCN, visual inspection revealed an obvious left-right asymmetry of c-Fos expression in the medial preoptic nucleus and subparaventricular zone of split hamsters killed during the inactive phase and in the medial division of the lateral habenula during the active phase (when the hamsters were running in their wheels). Roles for the dorsolateral SCN and the mediolateral habenula in circadian timekeeping are not yet understood. PMID- 16267384 TI - Are modifications of melatonin circadian rhythm in the middle years of life related to habitual patterns of light exposure? AB - The mechanisms underlying age-related changes in the signal from the biological clock have yet to be determined. The authors sought to determine if the phase advance of circadian melatonin rhythm during the middle years of life is related to different patterns of habitual light exposure. Forty-one healthy subjects between the ages of 22 and 58 y were studied. Habitual light exposure was measured by a wrist monitor for 7 days. Participants underwent a 25-h constant routine. They provided saliva samples every 30 min, and melatonin concentration was determined by radioimmunoassay to assess salivary dim light melatonin onset (S-DLMO(1.3)). Aging was associated with earlier S-DLMO(1.3). Increasing age was not related to the time spent at different light intensities. However, it was associated with lower percentage of light exposure during the night (between 0200 0400, 0600-0700, and 2300-2400 h) and with higher percentage of light exposure in the morning (between 0800-1100 h). Earlier S-DLMO(1.3) was associated with lower percentage of light exposure early on in the night (between 2200-0000, 0000-0100, and 0200-0300 h) as well as in the afternoon (between 1500-1600 h) and with higher percentage of light exposure in the morning (between 0800-1100 h). When the effects of age were controlled, there was no significant relationship between S-DLMO(1.3) and percentages of light exposure. Yet increasing age was associated with earlier S-DLMO(1.3) regardless of light exposure patterns. Earlier habitual wake time explained the earlier light exposure patterns of older subjects. Both habitual wake time and age contributed to the prediction of S-DLMO(1.3). The results suggest a phase advance of circadian rhythms in the middle years of life. Whereas a clear change in habitual light exposure patterns was associated with aging and with shifts in S-DLMO(1.3), it did not explain entirely the age-related advance of melatonin circadian phase. PMID- 16267385 TI - Shortened seasonal photoperiodic cycles accelerate aging of the diurnal and circadian locomotor activity rhythms in a primate. AB - The gray mouse lemur (Microcebus murinus), a prosimian primate, exhibits seasonal rhythms strictly controlled by photoperiodic variations. Previous studies indicated that longevity can be altered by long-term acceleration of seasonal rhythms, providing a model for assessing various aspects of aging. To assess the effect of aging and accelerated aging on the circadian system of this primate, we compared the circadian rhythm of the locomotor activity in adult mouse lemurs (2 4.5 years, n = 9), aged mouse lemurs (5-9 years, n = 10), and adult mouse lemurs that had been exposed from birth to a shortened seasonal photoperiodic cycle (2 4.5 years, n = 7). Compared to adult animals, aged mouse lemurs showed a significant increase in intradaily variability and an advanced activity onset. Aging was characterized by a decrease in amplitude, with both a decrease in nocturnal activity and an increase in daytime activity. When maintained in constant dim red light, aged animals exhibited a shortening of the free-running period (22.8 +/- 0.1 h) compared to adult animals (23.5 +/- 0.1 h). A 3- to 5 year exposure to an accelerated seasonal photoperiodic rhythm ("annual" duration of 5 months) in accelerated mouse lemurs produced disturbances of the locomotor activity rhythm that resembled those of aged mouse lemurs, whether animals were studied in entrained or in free-running conditions. The present study demonstrated a weakened and fragmented locomotor activity rhythm during normal aging in this primate. Increasing the number of expressed seasonal cycles accelerated aging of parameters related to circadian rhythmicity in adult animals. PMID- 16267386 TI - Expansion during primate radiation of a variable number tandem repeat in the coding region of the circadian clock gene period3. PMID- 16267388 TI - c-myc expression: keep the noise down! AB - The c-myc proto-oncogene encodes a nuclear protein that is deregulated and/or mutated in most human cancers. Acting primarily as an activator and sometimes as a repressor, MYC protein controls the synthesis of up to 10-15% of genes. The key MYC targets contributing to oncogenesis are incompletely enumerated and it is not known whether pathology arises from the expression of physiologic targets at abnormal levels or from the pathologic response of new target genes that are not normally regulated by MYC. Regardless of which, available evidence indicates that the level of MYC expression is an important determinant of MYC biology. The c-myc promoter has architectural and functional features that contribute to uniform expression and help to prevent or mitigate conditions that might otherwise create noisy expression. Those features include the use of an expanded proximal promoter, the averaging of input from dozens of transcription factors, and real time feedback using the supercoil-deformable Far UpStream Element (FUSE) as physical sensor of ongoing transcriptional activity, and the FUSE binding protein (FBP) as well as the FBP interacting repressor (FIR) as effectors to enforce normal transcription from the c-myc promoter. PMID- 16267389 TI - Regulation of macrophage ceruloplasmin gene expression: one paradigm of 3'-UTR mediated translational control. AB - Ceruloplasmin (Cp) is a copper protein with important functions in iron homeostasis and in inflammation. Cp mRNA expression is induced by interferon (IFN)-g in U937 monocytic cells, but synthesis of Cp protein is halted after about 12 h by transcript-specific translational silencing. The silencing mechanism requires binding of a 4-component cytosolic inhibitor complex, IFN gamma-activated inhibitor of translation (GAIT), to a defined structural element (GAIT element) in the Cp 3'-UTR. Translational silencing of Cp mRNA requires the essential proteins of mRNA circularization, suggesting that the translational inhibition requires end-to-end mRNA closure. These studies describe a new mechanism of translational control, and may shed light on the role that macrophage-derived Cp plays at the intersection of iron homeostasis and inflammation. PMID- 16267390 TI - Roles of heat shock protein gp96 in the ER quality control: redundant or unique function? AB - Heat shock protein gp96 is an endoplasmic reticulum chaperone, belonging to the HSP90 family. The function of gp96 as a molecular chaperone was discovered more than 10 years ago, but its importance has been overshadowed by the brilliance of its role in immune responses. It is now clear that gp96 is instrumental in the initiation of both the innate and adaptive immunity. Recently, the roles of gp96 in protein homeostasis, as well as in cell differentiation and development, are beginning to draw more attention due to rapid development in the structural study of HSP90 and some surprising new discoveries from genetic studies of gp96. In this review, we focus on the aspect of gp96 as an ER molecular chaperone in protein maturation, peptide binding and the regulation of its activity. PMID- 16267391 TI - Chromatin-remodeling factor INI1/hSNF5/BAF47 is involved in activation of the colony stimulating factor 1 promoter. AB - INI1/hSNF5/BAF47 is a core component of the hSWI/ SNF ATP-dependent chromatin remodeling complex, and it has been implicated in regulating gene expression, cell division and tumorigenesis. We investigated whether INI1/hSNF5/BAF47 functions in activation of the colony stimulating factor 1 (CSF1) promoter in HeLa cells. Overexpression of INI1/hSNF5/BAF47 promoted CSF1 transcription, and siRNA targeting INI1/hSNF5/ BAF47 (siINI1) strongly inhibited the activity of the CSF1 promoter. We demonstrated that all conserved domains of INI1/hSNF5/BAF47 are needed for CSF1 transcription. ChIP experiment showed that INI1/ hSNF5/BAF47 is recruited to the region of the CSF1 promoter. Taken together, these results indicate that INI1/hSNF5/BAF47 is involved in activation of the CSF1 promoter. PMID- 16267392 TI - Ethanol induces cell death by activating caspase-3 in the rat cerebral cortex. AB - Ethanol has long been implicated in triggering apoptotic neurodegeneration. We examined the effects of ethanol on the rat brain during synaptogenesis when a spurt in brain growth occurs. This period corresponds to the first 2 postnatal weeks in rats and is very sensitive to ethanol exposure. Ethanol was administered subcutaneously to 7-day- postnatal rat pups by a dosing regimen of 3 g/kg at 0 h and again at 2 h. Blood ethanol levels peaked (677+/-16.4 mg/dl) at 4 h after the first ethanol administration. The cerebral cortexes of the ethanol-treated group showed several typical symptoms of apoptosis such as chromosome condensation and disintegration of cell bodies. Activated caspase-3 positive cells were found in the cortex within 2 h of the first injection, and reached a peak at 12 h. In addition, TUNEL staining revealed DNA fragmentation in the same regions. These results demonstrate that acute ethanol administration causes neuronal cell death via a caspase-3-dependent pathway within 24 h, suggesting that activation of caspase-3 is a marker of the developmental neurotoxicity of ethanol. PMID- 16267393 TI - The Phosphorylation status of merlin is important for regulating the Ras-ERK pathway. AB - The neurofibromatosis type2 (NF2) tumor suppressor gene product, merlin, is structurally related to the ezrin-radixin-moesin (ERM) family of proteins that anchor the actin cytoskeleton to specific membrane proteins and participate in cell signaling. However, the basis of the tumor suppressing activity of merlin is not well understood. Previously, we identified a role of merlin as an inhibitor of the Ras-ERK signaling pathway. Recent studies have suggested that phosphorylation of merlin, as of other ERM proteins, may regulate its function. To determine whether phosphorylation of merlin affects its suppression of Ras-ERK signaling, we generated plasmids expressing full-length merlin with substitutions of serine 518, a potential phosphorylation site. A substitution that mimics constitutive phosphorylation (S518D) abrogated the ability of merlin to suppress effects of the Ras-ERK signaling pathway such as Ras-induced SRE transactivation, Elk-mediated SRE transactivation, Ras-induced ERK phosphorylation and Ras-induced focus formation. On the other hand, an S518A mutant, which mimics nonphosphorylated merlin, acted like wild type merlin. These observations show that mimicking merlin phosphorylation impairs not only growth suppression by merlin but also its inhibitory action on the Ras-ERK signaling pathway. PMID- 16267394 TI - An AFLP-based linkage map of Japanese red pine (Pinus densiflora) using haploid DNA samples of megagametophytes from a single maternal tree. AB - We have constructed an AFLP-based linkage map of Japanese red pine (Pinus densiflora Siebold et Zucc.) using haploid DNA samples of 96 megagametophytes from a single maternal tree, selection clone Kyungbuk 4. Twenty-eight primer pairs generated a total of 5,780 AFLP fragments. Five hundreds and thirteen fragments were verified as genetic markers with two alleles by their Mendelian segregation. At the linkage criteria LOD 4.0 and maximum recombination fraction 0.25(theta), a total of 152 markers constituted 25 framework maps for 19 major linkage groups. The maps spanned a total length of 2,341 cM with an average framework marker spacing of 18.4 cM. The estimated genome size was 2,662 cM. With an assumption of equal marker density, 82.2% of the estimated genome would be within 10 cM of one of the 230 linked markers, and 68.1% would be within 10 cM of one of the 152 framework markers. We evaluated map completeness in terms of LOD value, marker density, genome length, and map coverage. The resulting map will provide crucial information for future genomic studies of the Japanese red pine, in particular for QTL mapping of economically important breeding target traits. PMID- 16267395 TI - Characterization and transcriptional expression of the alpha-expansin gene family in rice. AB - The rice genome contains at least 28 EXPA (alpha-expansin) genes. We have obtained near full-length cDNAs from the previously uncharacterized genes. Analysis of these newly identified clones together with the 12 identified earlier showed that the EXPA genes contain up to two introns and encode proteins of 240 to 291 amino acid residues. The EXPA proteins contain three conserved motifs: eight cysteine residues at the N-terminus, four tryptophan residues at the C terminus, and a histidine-phenylalanine-aspartate motif in the central region. EXPA proteins could be divided into six groups based on their sequence similarity. Most were strongly induced in two-day-old seedlings and in the roots of one-week-old plants. However, only 14 genes were expressed in the aboveground organs, and their patterns were quite diverse. Transcript levels of EXPA7, 14, 15, 18, 21, and 29 were greater in stems, while EXPA2, 4, 5, 6, and 16 were highly expressed in both stem and sheath but not in leaf blade. EXPA1 is leaf blade-preferential, and EXP9 is leaf sheath-preferential. Most of the root expressed genes were more strongly expressed in the dividing zone. However, the Group 2 EXPA genes were also strongly expressed in both mature and dividing zones, while EXPA9 was preferentially expressed in the elongation zone. Fourteen EXPA genes were expressed in developing panicles, with some being expressed during most developmental stages, others only as the panicles matured. These diverse expression patterns of EXPA genes suggest that in general they have distinct roles in plant growth and development. PMID- 16267397 TI - Deficiency of Bloom's syndrome protein causes hypersensitivity of C. elegans to ionizing radiation but not to UV radiation, and induces p53-dependent physiological apoptosis. AB - Caenorhabditis elegans him-6 mutants, which show a high incidence of males and partial embryonic lethality, are defective in the orthologue of human Bloom's syndrome protein (BLM). When strain him-6(e1104) containing a missense him-6 mutation was irradiated with gamma-rays during germ cell development or embryogenesis, embryonic lethality was higher than in the wild type, suggesting a critical function of the wild type gene in mitotic and pachytene stage germ cells as well as in early embryos. Even in the absence of gamma-irradiation, apoptosis was elevated in the germ cells of the him-6 strain and this increase was dependent on a functional p53 homologue (CEP-1), suggesting that spontaneous DNA damage accumulates due to him-6 deficiency. However, induction of germline apoptosis by ionizing radiation was not significantly affected by the deficiency, indicating that HIM-6 has no role in the induction of apoptosis by exogenous DNA damage. We conclude that the C. elegans BLM orthologue is involved in DNA repair in promeiotic cells undergoing homologous recombination, as well as in actively dividing germline and somatic cells. PMID- 16267396 TI - Identification of genes differentially expressed in wild type and Purkinje cell degeneration mice. AB - Purkinje cell degeneration (pcd) mice are characterized by death of virtually all cerebellar Purkinje cells by postnatal day 30. In this study, we used DNA microarray analysis to investigate differences in gene expression between the brains of wild type and pcd mice on postnatal day 20, before the appearance of clear-cut phenotypic abnormalities. We identified 300 differentially expressed genes, most of which were involved in metabolic and physiological processes. Among the differentially expressed genes were several calcium binding proteins including calbindin-28k, paravalbumin, matrix gamma-carboxyglutamate protein and synaptotagamins 1 and 13, suggesting the involvement of abnormal Ca2+ signaling in the pcd phenotype. PMID- 16267398 TI - Effects of pine needle extract on pacemaker currents in interstitial cells of Cajal from the murine small intestine. AB - Extracts of pine needles (Pinus densiflora Sieb. et Zucc.) have diverse physiological and pharmacological actions. In this study we show that pine needle extract alters pacemaker currents in interstitial cells of Cajal (ICC) by modulating ATP-sensitive K+ channels and that this effect is mediated by prostaglandins. In whole cell patches at 30 degrees , ICC generated spontaneous pacemaker potentials in the current clamp mode (I = 0), and inward currents (pacemaker currents) in the voltage clamp mode at a holding potential of -70 mV. Pine needle extract hyperpolarized the membrane potential, and in voltage clamp mode decreased both the frequency and amplitude of the pacemaker currents, and increased the resting currents in the outward direction. It also inhibited the pacemaker currents in a dose-dependent manner. Because the effects of pine needle extract on pacemaker currents were the same as those of pinacidil (an ATP sensitive K+ channel opener) we tested the effect of glibenclamide (an ATP sensitive K+ channels blocker) on ICC exposed to pine needle extract. The effects of pine needle extract on pacemaker currents were blocked by glibenclamide. To see whether production of prostaglandins (PGs) is involved in the inhibitory effect of pine needle extract on pacemaker currents, we tested the effects of naproxen, a non-selective cyclooxygenase (COX-1 and COX-2) inhibitor, and AH6809, a prostaglandin EP1 and EP2 receptor antagonist. Naproxen and AH6809 blocked the inhibitory effects of pine needle extract on ICC. These results indicate that pine needle extract inhibits the pacemaker currents of ICC by activating ATP sensitive K+ channels via the production of PGs. PMID- 16267399 TI - PI3-kinase and PDK-1 regulate HDAC1-mediated transcriptional repression of transcription factor NF-kappaB. AB - PDK-1 activates PI3-kinase/Akt signaling and regulates fundamental cellular functions, such as growth and survival. NF-kB is involved in the induction of a variety of cellular genes affecting immunity, inflammation and the resistance to apoptosis induced by some anti-cancer drugs. Even though the crucial involvement of the PI3-kinase/Akt pathway in the anti-apoptotic activation of NF-kB is well known, the exact role of PDK-1 as well as PI3-kinase/Akt in NF-kB activation is not understood. Here we demonstrate that PDK-1 plays a pivotal role in transcriptional activation of NF-kB by dissociating the transcriptional co repressor HDAC1 from the p65 subunit of NF-kB. The association of CBP with p65 was not directly modulated by PDK-1 or by PI3-kinase. Etoposide activated NF-kB through PI3-kinase/Akt, and the transcription activation domain (TAD) of p65 was further activated by wild-type PDK-1. Overexpression of a dominant negative PDK-1 mutant decreased etoposide-induced NF-kB transcription and further down-regulated the ectopic HDAC1-mediated decrease in NF-kB transcriptional activity. Thus activation of PDK-1 relieves the HDAC1-mediated repression of NF-kB that may be related to basal as well as activated transcription by NF-kB. This effect may also explain the role of the PI3-kinase/PDK-1 pathway in the anti-apoptotic function of NF-kB associated with the chemoresistance of cancer cells. PMID- 16267400 TI - Differential expression of three catalase genes in hot pepper (Capsicum annuum L.). AB - Three different catalase cDNA clones (CaCat1, CaCat2, and CaCat3) were isolated from hot pepper (Capsicum annuum L.), and their expression patterns were analyzed at the levels of mRNA and enzyme activity. Northern hybridization showed that the three catalase genes were differentially expressed in various organs, and that expression of CaCat1 and CaCat2 was regulated differently by the circadian rhythm. In situ hybridization revealed different spatial distributions of CaCat1 and CaCat2 transcripts in leaf and stem. In response to wounding and paraquat treatment, CaCat1 mRNA increased at 4-12 h in both paraquat-treated and systemic leaves. In contrast, wounding had no significant effect on expression of the catalase genes. The increase of catalase activity in the paraquat-treated and systemic leaves paralleled that of CaCat1 mRNA, but did not match that of CaCat1 mRNA in paraquat-treated stems. Our results suggest that CaCat1 may play a role in responses to environmental stresses. PMID- 16267401 TI - Interaction of nonreceptor tyrosine-kinase Fer and p120 catenin is involved in neuronal polarization. AB - The neuronal cytoskeleton is essential for establishment of neuronal polarity, but mechanisms controlling generation of polarity in the cytoskeleton are poorly understood. The nonreceptor tyrosine kinase, Fer, has been shown to bind to microtubules and to interact with several actin-regulatory proteins. Furthermore, Fer binds p120 catenin and has been shown to regulate cadherin function by modulating cadherin-beta-catenin interaction. Here we show involvement of Fer in neuronal polarization and neurite development. Fer is concentrated in growth cones together with cadherin, beta-catenin, and cortactin in stage 2 hippocampal neurons. Inhibition of Fer-p120 catenin interaction with a cell-permeable inhibitory peptide (FerP) increases neurite branching. In addition, the peptide significantly delays conversion of one of several dendrites into an axon in early stage hippocampal neurons. FerP-treated growth cones also exhibit modified localization of the microtubule and actin cytoskeleton. Together, this indicates that the Fer-p120 interaction is required for normal neuronal polarization and neurite development. PMID- 16267402 TI - Angiotensin II promotes smooth muscle cell proliferation and migration through release of heparin-binding epidermal growth factor and activation of EGF-receptor pathway. AB - Transactivation of EGF-receptor (EGFR) by G-protein coupled receptors (GPCRs) is emerging as an important pathway in cell proliferation, which plays a crucial role in the development of atherosclerotic lesion. Angiotensin II (Ang II) has been identified to have a major role in the formation of atherosclerotic lesions, although the underlying mechanisms remain largely unclear. We hypothesize that Ang II promotes the proliferation and migration of smooth muscle cells through the release of heparin-binding epidermal growth factor like growth factor (HB EGF), transactivation of EGFR and activation of Akt and Erk 1/2, with matrix metalloproteases (MMPs) playing a dispensable role. Primary rat aortic smooth muscle cells were used in this study. Smooth muscle cells rendered quiescent by serum deprivation for 12 h were treated with Ang II (100 nM) in the presence of either GM6001 (20 microM), a specific inhibitor of MMPs or AG1478 (10 microM), an inhibitor of EGFR. The levels of phosphorylation of EGFR, Akt and Erk 1/2 were assessed in the cell lysates. Inhibition of MMPs by GM6001 significantly attenuated Ang II-stimulated phosphorylation of EGFR, suggesting that MMPs may be involved in the transactivation of EGFR by Ang II receptor. Furthermore Ang II stimulated proliferation and migration of smooth muscle cells were significantly blunted by inhibiting MMPs and EGFR and applying HB-EGF neutralization antibody, indicating that MMPs, HB-EGF and EGFR activation is necessary for Ang-II stimulated migration and proliferation of smooth muscle cells. Our results suggest that inhibition of MMPs may represent one of the strategies to counter the mitogenic and motogenic effects of Ang II on smooth muscle cells and thereby prevent the formation and development of atherosclerotic lesions. PMID- 16267403 TI - Alterations of protein expression in macrophages in response to Candida albicans infection. AB - Although macrophages are an important first line of cellular defense, they are unable to effectively kill phagocytosed C. albicans. To determine the physiological basis of this inability, we investigated the alterations of macrophage proteins caused by C. albicans infection. Since the formation of C. albicans hyphae caused cell death, proteins were prepared 3 h after infection and examined by two-dimensional gel electrophoresis (2-DE). The most prominent changes were in glycolytic enzymes, which could have caused energy depletion of the infected cells. Also changed were proteins involved in maintenance of cellular integrity and NO production. Treatment of the macrophages with either cytochalasin D or taxol did not alter their inability to kill C. albicans. Our results indicate that multiple factors contribute to cell death as the pathogenic form of C. albicans becomes fully active inside macrophage cells. PMID- 16267405 TI - Cytokine reporter mouse system for screening novel IL12/23 p40-inducing compounds. AB - Cytokines interleukin (IL) 12 and 23 play critical roles in linking innate and adaptive immune responses. They are members of heterodimeric cytokines, sharing a subunit p40. Although IL12/23 p40 is mainly induced in macrophages and dendritic cells (DCs) after stimulation with microbial Toll-like receptor ligands, methods to monitor the cells that produce IL12/23 p40 in vivo are limited. Recently, the mouse model to track p40-expressing cells with fluorescent reporter, yellow fluorescent protein, has been developed. Macrophages and DCs from these mice faithfully reported p40 induction using the fluorescent marker. Here we took advantage of these reporter mice to screen bio-compounds for p40-inducing activity. After screening hundreds of compounds, we found several extracts inducing IL12/23 p40 gene expression. Treatment of DCs with these extracts induced the expression of MHC class II and co-stimulatory molecules, which implies that these might be useful as adjuvants. Next, the in vivo target immune cells of candidate compounds were examined. The reporter system can be useful to identify cells producing IL12 or IL23 in vivo as well as in vitro. Thus, our cytokine reporter system proved to be a valuable reagent for screening for immunostimulatory molecules and identification of target cells in vivo. PMID- 16267404 TI - Insulin cannot activate extracellular-signal-related kinase due to inability to generate reactive oxygen species in SK-N-BE(2) human neuroblastoma cells. AB - The insulin-mediated Ras/mitogen-activated protein (MAP) kinase cascade was examined in SK-N-BE(2) and PC12 cells, which can and cannot produce reactive oxygen species (ROS), respectively. Tyrosine phosphorylation of the insulin receptor and insulin receptor substrate 1 (IRS-1) was much lower in SK-N-BE(2) cells than in PC12 cells when the cells were treated with insulin. The insulin mediated interaction of IRS-1 with Grb2 was observed in PC12 but not in SK-N BE(2) cells. Moreover, the activity of extracellular-signal-related kinase (ERK) was much lower in SK-N-BE(2) than in PC12 cells when the cells were treated with insulin. Application of exogenous H2O2 caused increased tyrosine phosphorylation and Grb2 binding to IRS-1 in SK-N-BE(2) cells, while exposure to an H2O2 scavenger (N-acetylcysteine) or to a phophatidylinositol-3 kinase inhibitor (wortmannin), and expression of a dominant negative Rac1, decreased the activation of ERK in insulin-stimulated PC12 cells. These results indicate that the transient increase of ROS is needed to activate ERK in insulin-mediated signaling and that an inability to generate ROS is the reason for the insulin insensitivity of SK-N-BE(2) cells. PMID- 16267408 TI - Adult-onset Still disease as the cause of fever of unknown origin. AB - We conducted the current study to evaluate the cases of fever of unknown origin (FUO) admitted in our institution during the 10 years between 1991 and 2001 and to compare the patients diagnosed as having adult-onset Still disease (AOSD) with the patients with FUO due to other diagnoses. We performed a case-control study and analyzed 26 patients with AOSD and 135 patients with FUO due to other diseases. Controls were classified into 1 of 4 groups: 1. Infectious diseases; 2. Malignant conditions; 3. Autoimmune diseases; 4. No diagnosis. Differences between groups were evaluated by analysis of variance (ANOVA). Odds ratios (OR) were calculated by multiple logistic regression analyses. Patients with AOSD were younger than controls. Arthritis (OR, 8.6; 95% confidence interval [CI], 1.5 49.1; p = 0.014), pharyngitis (OR, 6.9; 95% CI, 1.5-30.2; p = 0.010), splenomegaly (OR, 5.4; 95% CI, 1.1-26.7; p = 0.039), and neutrophilic leukocytosis (OR, 18.1; 95% CI, 3.5-93.6; p = 0.001) were significantly more common in patients with AOSD than in the control groups. A clinical scale that identifies patients with AOSD was designed. It proved to be highly specific ( approximately 98%), with predictive values greater than 90%.AOSD is a defined clinical entity. In most cases, it is clinically distinguishable from other causes of FUO. We propose a clinical scale as a tool to identify patients whose disease can be diagnosed based on clinical grounds without the need of long, costly diagnostic procedures. PMID- 16267406 TI - PLP-1 binds nematode double-stranded telomeric DNA. AB - The integrity and proper functioning of telomeres require association of telomeric DNA sequences with specific binding proteins. We have characterized PLP 1, a PURa homolog encoded by F45E4.2, which we previously identified as a candidate double stranded telomere binding protein, by affinity chromatography followed by mass spectrometry. PLP-1 bound double-stranded telomeric DNA in vitro as shown by competition assays. Core binding was provided by the third and fourth nucleotides of the TTAGGC telomeric repeat. This is quite different from the binding sequence of CEH-37, another C. elegans telomere binding protein, suggesting that multiple proteins may bind nematode telomeric DNA simultaneously in vivo. PMID- 16267409 TI - Immunogenetic risk and protective factors for the idiopathic inflammatory myopathies: distinct HLA-A, -B, -Cw, -DRB1 and -DQA1 allelic profiles and motifs define clinicopathologic groups in caucasians. AB - The idiopathic inflammatory myopathies (IIM) are systemic connective tissue diseases in which autoimmune pathology is suspected to promote chronic muscle inflammation and weakness. We have performed low to high resolution genotyping to characterize the allelic profiles of HLA-A, -B, -Cw, -DRB1, and -DQA1 loci in a large population of North American Caucasian patients with IIM representing the major clinicopathologic groups (n = 571). We confirmed that alleles of the 8.1 ancestral haplotype were important risk markers for the development of IIM, and a random forests classification analysis suggested that within this haplotype, HLA B*0801, DRB1*0301 and/ or closely linked genes are the principal HLA risk factors. In addition, we identified several novel HLA factors associated distinctly with 1 or more clinicopathologic groups of IIM. The DQA1*0201 allele and associated peptide-binding motif (KLPLFHRL) were exclusive protective factors for the CD8+ T cell-mediated IIM forms of polymyositis (PM) and inclusion body myositis (IBM) (pc < 0.005). In contrast, HLA-A*68 alleles were significant risk factors for dermatomyositis (DM) (pc = 0.0021), a distinct clinical group thought to involve a humorally mediated immunopathology. While the DQA1*0301 allele was detected as a possible risk factor for IIM, PM, and DM patients (p < 0.05), DQA1*03 alleles were protective factors for IBM (pc = 0.0002). Myositis associated with malignancies was the most distinctive group of IIM wherein HLA Class I alleles were the only identifiable susceptibility factors and a shared HLA-Cw peptide-binding motif (AGSHTLQWM) conferred significant risk (pc = 0.019). Together, these data suggest that HLA susceptibility markers distinguish different myositis phenotypes with divergent pathogenetic mechanisms. These variations in associated HLA polymorphisms may reflect responses to unique environmental triggers resulting in the tissue pathospecificity and distinct clinicopathologic syndromes of the IIM. PMID- 16267410 TI - Peripheral tuberculous lymphadenitis: epidemiology, diagnosis, treatment, and outcome. AB - We reviewed 106 patients referred to our institution for treatment of peripheral tuberculous adenitis to establish the epidemiologic, clinical, and pathologic manifestations of this disease. Tuberculous lymphadenitis occurred predominantly in young, foreign-born women a mean of 5 years after arrival in the United States. Tuberculin skin tests were positive in 94% of cases. Lymphadenopathy occurred most frequently in the neck (57%) or supraclavicular area (26%) and involved 1-3 nodes. Forty (38%) patients had an abnormal chest radiograph consistent with granulomatous infection. Culture-positive pulmonary tuberculosis was diagnosed in 41% of those patients with abnormal chest radiographs. Fine needle aspiration was an essential step in the evaluation and diagnosis of tuberculous lymphadenitis. Granulomas were seen in 61% of fine needle aspirates and 88% of surgical biopsies. Positive cultures for Mycobacterium tuberculosis were obtained from 62% of fine needle aspirate samples and 71% of excisional biopsies. The presence of necrosis and/or neutrophilic inflammation in tissue samples correlated with culture positivity. Given the high yield of positive cultures from fine needle aspirates, surgery was rarely indicated as an initial step in immunocompetent adults. In this cohort, 101 patients received a final diagnosis of peripheral tuberculous lymphadenitis. Eighty-two percent received their entire therapy under direct observation, and response to antituberculous therapy was uniformly successful. Paradoxical expansion of adenopathy was seen in 20% of all patients and was more commonly noted in human immunodeficiency virus seropositive patients. We present a diagnostic algorithm based on our experience. PMID- 16267411 TI - Outcome of sickle cell anemia: a 4-decade observational study of 1056 patients. AB - Based on a prospective cohort study of 1056 patients with sickle cell anemia (Hb SS) initiated in 1959, we investigated the influence of calendar era, age, sex, and prior medical conditions on the subsequent development of irreversible organ damage and survival using the Cox regression model with time-dependent covariates adjusting for all prior occurrences. We studied 30 acute clinical events, and focused on 8 prototypic forms of irreversible organ damage. Childhood survival to age 20 years has improved from 79% for those born before 1975 to 89% for children born in or after 1975. Bone infarction was a significant risk factor for avascular necrosis (p = 0.01), and infantile dactylitis was a significant risk factor for stroke (p = 0.01). Prior hospitalized vaso-occlusive sickle crisis in adults was significantly associated with the increased rate of avascular necrosis (p < 0.001), leg ulcers (p < 0.001), sickle chronic lung disease (p < 0.001), renal failure (p < 0.005), and early death (p < 0.001). The diagnosis of clearly evident clinical conditions such as leg ulcer, osteonecrosis, and retinopathy predicted an increased likelihood of developing a more lethal form of organ damage and earlier death: 77% of patients with chronic lung disease, 75% of those with renal insufficiency, and 51% of those with stroke had a prior chronic condition. Of the 232 patients who died, 73% had 1 or more clinically recognized forms of irreversible organ damage. By the fifth decade, nearly one-half of the surviving patients (48%) had documented irreversible organ damage. End-stage renal disease (glomerulosclerosis), chronic pulmonary disease with pulmonary hypertension, retinopathy, and cerebral microinfarctions are manifestations of arterial and capillary microcirculation obstructive vasculopathy. The current study underscores the need for preventive therapy to ameliorate the progression of the sickle vasculopathy. PMID- 16267412 TI - Rhabdomyolysis: an evaluation of 475 hospitalized patients. AB - Rhabdomyolysis is a common and potentially lethal clinical syndrome that results from acute muscle fiber necrosis with leakage of muscle constituents into blood. Myoglobinuria is the most significant consequence, leading to acute renal failure (ARF) in 15%-33% of patients with rhabdomyolysis. Rhabdomyolysis occurs from inherited diseases, toxins, muscle compression or overexertion, or inflammatory processes, among other disorders. In some cases, no cause is found. We describe 475 patients from the Johns Hopkins Hospital inpatient records between January 1993 and December 2001 for the following discharge diagnosis codes: myoglobinuria, rhabdomyolysis, myopathy, toxic myopathy, malignant hyperthermia, neuroleptic malignant syndrome, and polymyositis. Of 1362 patients, 475 patients with an acute neuromuscular illness with serum creatine kinase (CK) more than 5 times the upper limit of normal (>975 IU/L) were included. Patients with recent myocardial infarction or stroke were excluded. The etiology was assigned by chart review. For all, the highest values of serum CK, serum creatinine and urine myoglobin, hemoglobin, and red blood cells were recorded. Forty-one patients had muscle biopsy within at least 2 months from the onset of rhabdomyolysis.Of the 475 patients, 151 were female and 324 were male (median age, 47 yr; range, 4-95 yr). Exogenous toxins were the most common cause of rhabdomyolysis, with illicit drugs, alcohol, and prescribed drugs responsible for 46%. Among the medical drugs, antipsychotics, statins, zidovudine, colchicine, selective serotonin reuptake inhibitors, and lithium were the most frequently involved. In 60% of all cases, multiple factors were present. In 11% of all cases, rhabdomyolysis was recurrent. Underlying myopathy or muscle metabolic defects were responsible for 10% of cases, in which there was a high percentage of recurrence, only 1 etiologic factor, and a low incidence of ARF. In 7%, no cause was found. ARF was present in 218 (46%) patients, and 16 died (3.4%). A linear correlation was found between CK and creatinine and between multiple factors and ARF, but there was no correlation between ARF and death or between multiple factors and death. Urine myoglobin detected by dipstick/ultrafiltration was positive in only 19%. Toxins are the most frequent cause of rhabdomyolysis, but in most cases more than 1 etiologic factor was present. Patients using illicit drugs or on prescribed polytherapy are at risk for rhabdomyolysis. The absence of urine myoglobin, by qualitative assay, does not exclude rhabdomyolysis. With appropriate care, death is rare. PMID- 16267413 TI - Ear myxomas in Carney's complex. PMID- 16267414 TI - Malakoplakia of the neck in an immunosuppressed patient. PMID- 16267415 TI - Ipsilateral replantation of foot with crossover segmental transfer in bilateral leg amputation. PMID- 16267416 TI - Gender identity disorder: general overview and surgical treatment for vaginoplasty in male-to-female transsexuals. AB - LEARNING OBJECTIVES: After studying this article, the participant should be able to discuss: 1. The terminology related to male-to-female gender dysphoria. 2. The different theories regarding cause, epidemiology, and treatment of gender dysphoria. 3. The surgical goals of sex reassignment surgery in male-to-female transsexualism. 4. The surgical techniques available for sex reassignment surgery in male-to-female transsexualism. BACKGROUND: Gender identity disorder (previously "transsexualism") is the term used for individuals who show a strong and persistent cross-gender identification and a persistent discomfort with their anatomical sex, as manifested by a preoccupation with getting rid of one's sex characteristics, or the belief of being born in the wrong sex. Since 1978, the Harry Benjamin International Gender Dysphoria Association (in honor of Dr. Harry Benjamin, one of the first physicians who made many clinicians aware of the potential benefits of sex reassignment surgery) has played a major role in the research and treatment of gender identity disorder, publishing the Standards of Care for Gender Dysphoric Persons. METHODS: The authors performed an overview of the terminology related to male-to-female gender identity disorder; the different theories regarding cause, epidemiology, and treatment; the goals expected; and the surgical technique available for sex reassignment surgery in male-to-female transsexualism. RESULTS: Surgical techniques available for sex reassignment surgery in male-to-female transsexualism, with advantages and disadvantages offered by each technique, are reviewed. Other feminizing nongenital operative interventions are also examined. CONCLUSIONS: This review describes recent etiopathogenetic theories and actual guidelines on the treatment of the gender identity disorder in male-to-female transsexuals; the penile-scrotal skin flap technique is considered the state of the art for vaginoplasty in male-to-female transsexuals, whereas other techniques (rectosigmoid flap, local flaps, and isolated skin grafts) should be considered only in secondary cases. As techniques in vaginoplasty become more refined, more emphasis is being placed on aesthetic outcomes by both surgeons and patients. PMID- 16267417 TI - The conjoint medial circumflex femoral perforator and gracilis muscle free flap: anatomical study and clinical use for complex facial paralysis reconstruction. AB - BACKGROUND: [corrected] The aim of this study was to establish the anatomic basis of the conjoint medial circumflex femoral perforator and gracilis muscle flap and to expand the use of this flap in complex facial paralysis reconstruction. METHODS: An anatomic study was initially undertaken to record the existence, consistency, and diameter of musculocutaneous perforators emanating from the proximal third of the gracilis muscle to provide blood supply to the overlying fascia, subcutaneous fat, and skin. In a total of 20 clinical cases of gracilis muscle harvesting, the aforementioned anatomical data were recorded during flap dissection. At least one musculocutaneus perforator, consisting of one artery and two accompanying veins (vein caliber > 0.3 mm) was found in 95 percent of cases. RESULTS: The anatomical study was followed by successful use of the conjoint flap for reconstruction of longstanding facial palsy accompanied by a soft-tissue defect of the cheek. In the first stage, cross-face nerve grafting was performed. In the second stage, free transfer of the conjoint flap, consisting of the proximal third of the gracilis muscle and the overlying subcutaneous fat, was performed to the face. The only connection between the two components of the conjoint flap was one musculocutaneous perforator. When the flap was inset, the muscle was used for facial reanimation and partial obliteration of the soft tissue defect, while the subcutaneous fat was used to obliterate the rest of the defect. CONCLUSION: The proposed technique ensured symmetry of the face, on both rest and animation, and obliteration of the cheek deformity. PMID- 16267418 TI - Gunshot wounds: reconstruction of the lower face by osteogenic distraction. AB - BACKGROUND: Although osteogenic distraction is a well-established technique, the distraction device still needs to be improved, miniaturized, and made lighter, more flexible, and more adaptable for mandibular reconstruction in adults with gunshot wounds. The authors successively used unidirectional and bidirectional devices, followed by a bone transporter with a horseshoe-shaped trammel. The trammel system was then replaced by an endless screw, and finally by a customized endless screw. METHODS: Eleven adult patients with gunshot injuries underwent mandibular reconstruction using osteogenic distraction with an external device. RESULTS: An average bone gain of 79 mm was achieved. No infectious complications were observed. The authors encountered equipment problems during the study, requiring a change of material. The mean duration of mandibular distraction was 3.5 months. CONCLUSIONS: Distraction of bone fragments facilitates the simultaneous expansion of soft tissues, avoiding free or pedicled myocutaneous flaps, for soft-tissue reconstruction. The alveolar ridge with the attached gum is also recreated by distraction, allowing dental reconstruction by osseointegrated implants. PMID- 16267419 TI - Island rotation flap for nasal reconstruction. AB - BACKGROUND: Nasal reconstruction options are often dictated by size and location of the defect. Local flaps are often considered of little utility for larger or more complex defects. The authors' modification of the island rotation flap allows increased versatility and single-stage reconstruction for many more defects of varied sizes and locations. METHODS: A retrospective review of all cases of nasal reconstruction using the island rotation flap over a 3.5-year period was undertaken. RESULTS: Ninety-two island flap reconstructions were performed for defects ranging from 0.8 to 4.0 cm. For defects smaller than 2.5 cm, there were no cases of flap loss. Five percent of the patients required a second procedure, whereas 95 percent were reconstructed in a single stage. Five percent required treatment for prominent scars. Follow-up was over 2 years. CONCLUSIONS: The laterally based blood supply of the nasal island rotation flap is reliable and robust. Patients treated with this flap had good healing and good symmetry and required minimal revision. Single-stage reconstruction with this flap requires both proper flap design and extensive undermining of the entire nasal soft-tissue cover. Nasal symmetry is maintained despite the defect and donor flap being on the same side of the nose. The authors present their technique for the island rotation flap for nasal reconstruction and information on its reliability and acceptance. PMID- 16267421 TI - Combining the Cutting and Mulliken methods for primary repair of the bilateral cleft lip nose. AB - BACKGROUND: Since 1990, primary bilateral cleft nasal reconstruction has been focused on placing the lower lateral cartilages into normal anatomical position. Of the four major techniques in this class, the Cutting (i.e., retrograde) method and the Mulliken method have been most successful. The retrograde method makes no external nasal incisions, but requires either preoperative or postoperative nasal molding to achieve maximum benefit. Mulliken's technique does not require molding, but leaves the footplates of the medial crura in the depression above the projecting premaxilla associated with the diminutive anterior nasal spine. Leaving the footplates in place also prevents adequate approximation of the alar bases. In this article, the two methods are combined to achieve the benefits of both. METHODS: We report our experience with the retrograde nasal approach associated with marginal rim incisions (Mulliken method) in a series of 25 consecutive bilateral cleft lip cases simultaneous with lip repair. We performed a retrograde approach through membranous septum incisions elevating a prolabial columellar flap. To facilitate alar cartilage manipulation we added bilateral marginal rim incisions. Nasal width, columella length and width, tip projection, and nasolabial angle were analyzed after a minimum of 2 years after surgery. These were compared with a normal, age-matched, control group. We also examined nostril symmetry and marginal nostril scars. RESULTS: Columellar length was not statistically significantly different from that of the control group (p = 0.122442). Nasal width, columellar width, tip projection, and nasolabial angle were all significantly greater in the cleft group than normal (p < 0.001). No hypertrophied scars were found associated with the marginal rim scar. CONCLUSIONS: Adding the Mulliken approach allows alar cartilage manipulation to be performed more easily than when using the retrograde approach alone. Tip projection and alar base narrowing are facilitated using the combined technique rather than the Mulliken approach alone. Prolabial flap manipulation is safe using this combined approach, even in cases with a severely projected premaxilla. We believe that the combined approach is safe and yields better long-term results than either technique alone. PMID- 16267423 TI - Double unilimb Z-plastic repair of microform cleft lip. AB - BACKGROUND: Microform unilateral cleft lip is characterized by 1) notched mucosal margin; 2) thin medial vermilion; 3) elevated medial peak of Cupid's bow; 4) furrowed philtral column; 5) hypoplastic orbicularis oris; and 6) minor nasal deformity. METHODS: The author's registry of unilateral incomplete cleft lip was culled for patients with microform cleft lip. Operative correction included: double-limb Z-plasty at the vermilion-cutaneous and vermilion-mucosal junctions; eversion of orbicularis oris; augmentation of philtral ridge with a dermal graft; medial positioning of the alar base; and elevation of the lower lateral cartilage. RESULTS: Microform phenotype was found in 33 of 360 infants (9.2 percent) with unilateral incomplete cleft lip. Male-to-female and left-to-right ratio were both 2:1. Median age at presentation was 11 months (range, 2 weeks to 9 years). Twenty-three patients had a double unilimb Z-plastic repair (including dermal graft and nasal correction). No revisions have been necessary at median follow-up of 5 years, however, 13 percent of children lacked prominence of the upper philtral column and one-third of children exhibited minor nostril asymmetry. CONCLUSIONS: Double unilimb Z-plasty corrects the vertical asymmetry in a microform cleft lip while limiting the scar to the lower one-half of the lip. The philtral ridge is formed by repair of the muscular diastasis and onlay of a dermal graft. Components of this technique are applicable to secondary cleft deformities, such as elevated peak of the Cupid's bow and inadequate philtral ridge. PMID- 16267424 TI - Maximizing outcomes in breast reduction surgery: a review of 518 consecutive patients. AB - BACKGROUND: Economic constraints and diminished healthcare resources mandate increased efficiency in labor-intensive plastic surgical procedures, such as reduction mammaplasty. The evolution to our current approach over a 9-year period was reviewed. METHODS: From 1992 to 2001 a total of 518 patients underwent bilateral reduction mammaplasty by a single plastic surgeon. Since 1992, a bilateral simultaneous approach to reduction mammaplasty was used, with the primary surgeon (Scott) performing the preoperative markings and determining the final resection. An inferior pedicle, "inverted T" technique using a Wise pattern was used for all cases. The initial 2-year period (113 consecutive patients from 1992 to 1994) was compared with the later 2-year period (103 patients from 1999 to 2001). Variables that decreased operative times and resources were identified. RESULTS: Length of stay (from admission to discharge) was initially 27 hours (overnight admission at a hospital) in the early review and decreased to 5 hours (outpatient surgery center) in the recent series. Drains were used in 100 percent of the earlier cases and in 7 percent in the recent series. Major complications occurred in 3 percent of the entire series. Minor complications occurred in 20 percent of all patients. In the recent series 97 percent of patients received complete relief of their preoperative symptoms and expressed satisfaction with their results. Operative times decreased from 120 to 102 minutes. Beginning in 1999, Dermabond skin adhesive was used in all patients. CONCLUSIONS: An evolutionary approach to bilateral reduction mammaplasty with low complication rates and high patient satisfaction is presented. Optimization of technique leads to decreased operative times, shortened lengths of stay, and contributes to efficient use of surgeon and operating room resources. PMID- 16267426 TI - Unilateral postoperative chest wall radiotherapy in bilateral tissue expander/implant reconstruction patients: a prospective outcomes analysis. AB - BACKGROUND: Implant reconstruction has a major role to play in breast reconstruction, as some patients neither wish nor are suitable for autogenous reconstruction. The suitability of implant reconstruction in patients who may receive postoperative, adjuvant radiation therapy has not, however, been fully clarified. The purpose of this study was to evaluate complications, capsular contracture, aesthetic outcomes, and patient satisfaction in patients who have undergone bilateral tissue expander/implant reconstruction and unilateral, post exchange, adjuvant radiotherapy. In this study population, the effect of radiation can best be appreciated because the nonirradiated breast acts as a control. METHODS: A review of all bilateral tissue expander/implant reconstructions at a single cancer center was undertaken. Twelve patients who underwent bilateral expander/implant reconstruction and unilateral postexchange radiotherapy were eligible for participation. A prospective evaluation of complications, cosmesis, and patient satisfaction was performed. The evaluation of cosmesis and patient satisfaction was executed for 10 of the 12 patients, as two were dead at the time of follow-up. RESULTS: Mean follow-up was 23.5 months (range, 12 to 58.5 years). In 40 percent of patients, there was no discernible difference in capsular contracture between the irradiated and nonirradiated breasts. In 50 percent of patients, the irradiated breast demonstrated increased contracture by a single modified Baker grade. In 10 percent of patients, contracture of the irradiated breast was two modified Baker grades greater than that of the nonirradiated side (grade III versus grade I). CONCLUSION: For the majority of patients, the degree of capsular contracture was higher on the irradiated side, yet overall symmetry, aesthetic results, and patient satisfaction remained high. These data support the conclusion that immediate, bilateral breast reconstruction using tissue expansion and implants is an acceptable option for the subset of patients who may undergo unilateral, postexchange radiotherapy. PMID- 16267427 TI - A randomized, controlled trial to determine the efficacy of paper tape in preventing hypertrophic scar formation in surgical incisions that traverse Langer's skin tension lines. AB - BACKGROUND: How a scar is managed postoperatively influences its cosmetic outcome. After suture removal, scars are susceptible to skin tension, which may be the trigger for hypertrophic scarring. Paper tape to support the scar may reduce multidirectional forces and prevent hypertrophic scarring. METHODS: Seventy patients who had undergone cesarean section at the Royal Brisbane and Women's Hospital were randomized to treatment and control groups. Patients in the control group received no postoperative intervention. Patients in the treatment group applied paper tape to their scars for 12 weeks. Scars were assessed at 6 weeks, 12 weeks, and 6 months after surgery using ultrasound to measure intradermal scar volume. Scars were also assessed using the International Clinical Recommendations. RESULTS: Paper tape significantly decreased scar volume by a mean of 0.16 cm3, (95 percent confidence interval, 0.00 to 0.29 cm3). At 12 weeks after surgery, 41 percent of the control group developed hypertrophic scars compared with none in the treatment group (exact test, p = 0.003). In the treatment group, one patient developed a hypertrophic scar and four developed stretched scars only after the tape was removed. The odds of developing a hypertrophic scar were 13.6 times greater in the control than in the treatment group (95 percent confidence interval, 3.6 to 66.9). Of the 70 patients randomized, 39 completed the study. Four patients in the treatment group developed a localized red rash beneath the tape. These reactions were minor and transient and resolved without medical intervention. CONCLUSIONS: The development of hypertrophic and stretched scars in the treatment group only after the tape was removed suggests that tension acting on a scar is the trigger for hypertrophic scarring. Paper tape is likely to be an effective modality for the prevention of hypertrophic scarring through its ability to eliminate scar tension. PMID- 16267429 TI - Free tissue transfer in the elderly: incidence of perioperative complications following microsurgical reconstruction of 197 septuagenarians and octogenarians. AB - BACKGROUND: There are an increasing number of elderly patients requiring free tissue transfer. The risks of complex reconstructions in this patient population remain largely unknown. Therefore, the authors' reason for conducting this study was to review their experience with free tissue transfer in patients aged 70 years or older. METHODS: A retrospective review of all free tissue transfers performed over a 10-year period at Memorial Sloan-Kettering Cancer Center was performed and all patients aged 70 years or older were identified. Medical records and the authors' prospectively maintained database were analyzed with respect to comorbidities and postoperative complications. RESULTS: Two hundred eleven patients aged 70 years or older (70 to 79 years, n = 184; 80+ years, n = 27) were identified during the study period. Of these, 197 (70 to 79 years, n = 170; 80+ years, n = 27) patients had complete charts for review. Flap survival was 100 percent in the 80+ group and 97 percent in the 70 to 79 group. The overall complication rate was 59.3 percent in the 80+ group and 35.3 percent in the 70 to 79 group (p = 0.030). The medical complication rate was 40.7 percent in octogenarians and 11.8 percent in septuagenarians (p = 0.0004). Overall surgical complications were similar in the two groups. Univariate analysis demonstrated that age was associated with medical complications but not surgical complications. Using multivariate analysis, the authors found that alcohol use and coronary artery disease were independent predictors of overall, medical, and surgical complications. CONCLUSIONS: Free tissue transfer may be performed in patients over age 70 with a high degree of technical success. The procedure, however, carries a distinct risk of perioperative mortality and morbidity, particularly in patients over the age of 80. Comorbidities significantly associated with complications include age, alcohol use, coronary disease, and hypertension. This study suggests that prolonged survival may be achieved in some patients; however, a selective approach is required. PMID- 16267431 TI - Chronic asymptomatic contralateral wrist scapholunate dissociation. AB - BACKGROUND: The authors report a series of six patients who presented with scapholunate dissociation, with no significant contralateral antecedent trauma and no rheumatoid arthritis or congenital ligamentous laxity. Such patients may present with pain or a snapping/popping sensation in one wrist, undergo radiographs of both wrists, and are discovered to have scapholunate gapping bilaterally. The literature contains few reports of this condition, and this series of six is a relatively large one for this infrequently reported condition. METHODS: The six charts were reviewed retrospectively; each patient was asked to return for follow-up and radiographic examination and each participated in a telephone questionnaire about pain, activity changes, new treatments, and exacerbation of wrist problems. The average follow-up was 39 months, with a range of 6 months to 13.5 years. RESULTS: One patient with severe unilateral instability and persistent pain underwent soft-tissue surgical repair (Blatt reconstruction); another demonstrated unilateral dorsal intercalated segment instability with moderate pain symptoms but declined surgical reconstruction. The other 10 wrists, despite radiographically demonstrated widened scapholunate angles and rotatory subluxation of the scaphoid, had mild or no pain and no dorsal intercalated segment instability deformity. CONCLUSIONS: The evolution of the bilateral form of scapholunate dissociation seems to be benign unless dorsal intercalated segment instability deformity is present, which may then rapidly progress to degenerative arthritis and scapholunate advanced collapse wrist. Severe or minor repetitive trauma, inflammation, infection, tumors, and congenital ligamentous laxity have been etiologically implicated in scapholunate dissociation. The natural history of scapholunate dissociation involves volar rotation of the scaphoid and dorsal rotation of the lunate, progressing to malalignment and eventual arthrosis between the scaphoid and radius, the capitate and lunate, and the lunate and hamate bones. PMID- 16267432 TI - Glabrous dermal grafting: a 12-year experience with the functional and aesthetic restoration of palmar and plantar skin defects. AB - BACKGROUND: Glabrous skin on the palmar aspect of the hands and the plantar aspect of the feet has special attributes. These attributes define the skin on the palm, fingers, and sole as functionally and aesthetically different from skin on other parts of the body. When there is a glabrous skin defect, it should be replaced with similar skin to restore function and aesthetics. The authors report their 12-year experience with the technique of glabrous dermal grafting for the reconstruction of palmar and plantar skin defects. METHODS: From 1992 to 2004, 13 patients with 14 defects underwent glabrous dermal grafting of either palmar or plantar defects. Defects included nine hand and five foot defects. Causes included nine acute burns, one secondary burn reconstruction, two delayed reconstructions of traumatic injuries, one congenital nevus, and one malignant melanoma. Donor sites included 12 glabrous dermal grafts from the foot and two from the hand. RESULTS: Follow-up ranged from 1 month to 65 months. All glabrous dermal grafts demonstrated complete epithelialization and no incidence of complete loss. There was return of sensation without hyperkeratosis or breakdown. The grafts demonstrated good color match with the surrounding skin. The donor site healed without complications, and there were no incidences of significant hypopigmentation, hyperpigmentation, or hypertrophic scarring. CONCLUSION: Glabrous dermal grafting of palmar and plantar defects is the ideal way of reconstructing glabrous skin to restore both function and aesthetics and minimize donor-site morbidity. PMID- 16267433 TI - The osteogenic potential of adipose-derived mesenchymal cells is maintained with aging. AB - BACKGROUND: Adipose-derived mesenchymal cells are multipotent progenitor cells derived from the vascular-stromal compartment of adipose tissue. Although we have recently shown that these cells, from both juvenile and adult animals, are capable of forming bone in vivo, a detailed examination of the differences in the biology of these two populations (and in particular their ability to form bone) has not been performed. METHODS: Adipose-derived mesenchymal cells were harvested from juvenile (6-day-old) and adult (60-day-old) mice. Differences in cellular attachment, proliferation, and proliferating cell nuclear antigen production were assessed. The ability of cells to undergo adipogenic differentiation was determined by Oil Red O staining. Early osteogenic differentiation was determined with alkaline phosphatase staining, and terminal differentiation with von Kossa staining as well as determination of extracellular matrix calcium content. All experiments were performed in triplicate. RESULTS: Greater attachment, proliferation, and proliferating cell nuclear antigen production were seen in juvenile as compared with adult adipose-derived mesenchymal cells. The juvenile cells underwent significantly greater adipogenic differentiation than did adult cells (p < 0.001). Interestingly, the adult cells were capable of robust early and terminal osteogenic differentiation, with levels of all three osteo-genic assays being similar to those seen in juvenile cells. Differences were not statistically significant. CONCLUSIONS: Although biologic differences exist between adipose-derived mesenchymal cells from juveniles and adults, the osteogenic capacity of these cells appears to be minimally affected by donor age. This suggests that these cells may be a particularly useful cellular resource in the design of cell-based therapies for skeletal regeneration in an aging population. PMID- 16267434 TI - Nerve regeneration through a healthy nerve trunk: a new and hopeful conduit for bridging nerve defects. AB - BACKGROUND: Considering a healthy nerve trunk as the hypothetically ideal conduit, a new experimental model using an intact nerve for bridging a nerve defect was contemplated. METHODS: Thirty rats were used. In group I (double coaptation), a segment was removed from the peroneal nerve. Both the proximal and distal stumps were repaired end-to-side to the tibial nerve. In group II (only distal coaptation), only the distal nerve stump was repaired. In group III (control), the transected segment was immediately repaired primarily in its original orientation as a nerve graft. A walking track analysis was conducted periodically for 28 months. The horseradish peroxidase retrograde labeling technique was used for tracking the origin of the axons presented in the distal stump of the peroneal nerve in group I, and morphologic studies were also carried out for all the groups. RESULTS: Functional assessment revealed that the difference between group I and group II was significant. The horseradish peroxidase labeling test suggested that the nerve fibers in the distal stump of the peroneal nerve were mostly from its original proximal stump passed by the way of the tibial nerve bridge. CONCLUSION: This study suggested that the axons of the proximal stump of a sectioned nerve can sprout into another intact nerve trunk by the way of an end-to-side repair site, regenerate, and advance in its epineurium distally for a distance and pass into its original distal stump if it was repaired end-to-side. It was thought that the technique could be used in clinical cases with short nerve defects as an alternative method to grafts and conduits. PMID- 16267435 TI - Treatment of severely contracted fingers with combined use of cross-finger and side finger transposition flaps. AB - BACKGROUND: In the present study, the authors combined use of the cross-finger flap and the side-finger transposition flap to cover the skin and soft-tissue defect created by contracture release of severely contracted fingers. METHODS: Eight patients having Stern type III flexion contractures of the proximal interphalangeal joints were included. The cause of injury was burn in six patients and trauma in two patients. The average follow-up period was 11.6 months. RESULTS: All operations were successful. Lack of extension of the proximal interphalangeal joint was improved by approximately 81.2 degrees for all digits. CONCLUSION: Stern type III contracture of the proximal interphalangeal joint can be released by transverse incision and ample resection of scarred tissue, and the resulting palmar skin defect that cannot be covered by using the finger's own flaps or cross-finger flap can be covered by combined use of cross finger and side-finger transposition flaps. PMID- 16267436 TI - Use of the omentum flap as additional soft-tissue cover for abdominal wall defects reconstructed with Gore-Tex. AB - BACKGROUND: Use of alloplastic materials such as Gore-Tex and locoregional flaps for reconstruction of large abdominal wall defects has been well described. The purpose of this article is to present a novel technique of using the omentum as an interpositional flap to protect the Gore-Tex repair of the abdominal wall. METHODS: Four patients with large abdominal wall defects underwent reconstruction with Gore-Tex and omentum flap. These defects resulted from tumor resection and recurrent incisional hernia. Their dimensions ranged from 15 x 10 cm to 25 x 27 cm. The Gore-Tex patch was inset using an underlay technique. The omentum was tunneled through a separate opening in the abdominal wall into the subcutaneous plane and used to cover the Gore-Tex. Skin coverage was accomplished by direct closure or myocutaneous flaps. RESULTS: The mean follow-up was 17 months. All wounds healed, with no hernias. One patient developed a subcutaneous abscess 6 months postoperatively, and this was treated successfully by percutaneous drainage, as the omentum had walled-off the abscess. CONCLUSION: The omentum flap served as an additional soft-tissue cover over the Gore-Tex repair to prevent exposure in the event of infection or flap breakdown. This technique is useful in situations in which delayed wound healing is anticipated or when large quantities of prosthetic material are used. PMID- 16267437 TI - A simple, inexpensive nerve approximator. PMID- 16267438 TI - Further clinical experience with synthetic mesh for the entire abdominal wall after TRAM flap breast reconstruction. PMID- 16267439 TI - New frontiers in calvarial reconstruction: integrating computer-assisted design and tissue engineering in cranioplasty. AB - Repair of large and complex calvarial defects remains a particular challenge for reconstruction. The paucity of techniques and materials emphasizes the need for alternative bone formation strategies. Recent integrative approaches suggest that successful reconstruction requires interdisciplinary teams, with surgeons interacting with imaging experts, materials scientists, and engineers. In this review, the authors present an overview of current materials used in calvarial reconstruction. Subsequently, progress in computer-designed prostheses, tissue engineering, and osteoinduction strategies is discussed. Finally, the authors discuss their experience with the integration of computer-aided fabrication of customized implants and tissue engineering for calvarial reconstruction. PMID- 16267440 TI - Analysis of the nerve branches to the orbicularis oculi muscle of the lower eyelid in fresh cadavers. AB - BACKGROUND: Scleral show or ectropion is a known complication of lower eyelid surgery. It is particularly problematic after lower eyelid blepharoplasty when the orbicularis oculi muscle is transected or after other procedures are performed that denervate the lower eyelid. The purpose of this study was to determine whether a dominant motor branch exists to the lower lid orbicularis oculi muscle, using anatomic dissection and histologic analysis. METHODS: Sixteen fresh facial cadaver halves were dissected using the operative microscope to identify and measure nerve branches. The nerves were then harvested, sectioned, and stained with toluidine blue dye for histomorphometric analysis. Nerves were categorized as either lateral or medial to a vertical line marked at the lateral edge of the limbus of the eye. RESULTS: Eighty-seven percent and 47 percent of cadaver halves had one and two nerve branches lateral to the limbus, respectively. Eighty percent and 47 percent of cadaver halves had one and two nerve branches medial to the lateral limbus, respectively. The nerve branches entered the inferior edge of the orbicularis oculi muscle between 0.88 cm (+/ 0.36 cm) and 2.73 cm (+/-0.46 cm) from the lateral canthus. There was no correlation noted between fascicular counts and distance from the lateral canthus. CONCLUSIONS: Multiple motor nerves exist to the lower eyelid. Branches are found medial and lateral to the lateral limbus, and no nerve branch is dominant. PMID- 16267442 TI - A key to canthopexy: the tarsal strap. A fresh cadaveric study. AB - BACKGROUND: This article describes a poorly understood yet clinically significant eyelid structure the authors have labeled the tarsal strap. The tarsal strap anchors the tarsus to the periosteum of the inferolateral orbit. Releasing this tethering band is the cornerstone of achieving lasting, superior surgical results during lateral canthopexy and periorbital restoration in aesthetic and reconstructive surgery. METHODS: An 87-year-old, male, fresh cadaveric head of Caucasian origin was studied. The dissection was repeated on 15 cadaver heads, including both male and female cadavers, from various ethnic backgrounds, to confirm the anatomy of the tarsal strap described in this article. RESULTS: The tarsal strap was identified in all 16 cadaver dissections and appeared in a consistently similar location. CONCLUSIONS: Until now, the tarsal strap has been a poorly understood entity, despite being an anatomically and clinically significant eyelid structure. Knowledge of its significance is essential to restoring the lateral canthus and returning periorbital structures to their youthful state. It should be routinely sought out and properly released. PMID- 16267444 TI - Brow fixation with the Endotine Forehead device in endoscopic brow lift. AB - BACKGROUND: The Endotine Forehead device (Coapt Systems, Inc., Palo Alto, Calif.) is an implantable bioabsorbable fixation device designed to provide intuitive, multipoint, distributed tension and repeatable and predictable brow fixation during endoscopic and open browplasty. The purpose of this study was to evaluate early results in a series of endoscopic brow lift cases using the Endotine Forehead device. METHODS: Two versions of the Endotine device were used in this study. One was composed of polylactic acid, and the other was a smaller version consisting of 82/18 L-lactide/glycolide). In a consecutive series of endoscopic brow lift cases, preoperative and postoperative standardized photographs were taken in the Frankfort horizontal plane and three measurements were compared: midpupil to superior brow, midpupil to hairline, and lateral canthus to superior brow. RESULTS: A total of 21 patients (19 women and two men) underwent endoscopic browplasty. Photographs were obtained preoperatively and 54 to 174 days postoperatively. Brow elevation measurements were obtained postoperatively. No significant adverse events were encountered in the follow-up period. CONCLUSION: The Endotine Forehead device provides significant and reproducible brow elevation, with no significant adverse events, as measured at three points in excess of 14 weeks postoperatively. PMID- 16267446 TI - Visible scars from percutaneous osteotomies. AB - BACKGROUND: One of the criticisms of the percutaneous lateral nasal osteotomy technique is the potential for unfavorable, visible scarring. No clinical study has compared the external percutaneous approach with an internal approach to evaluate scarring in the same patient. This prospective, randomized, blinded study was designed to test the hypothesis that percutaneous perforating osteotomies cause perceptible scars when compared with the transnasal internal lateral osteotomy methods. METHODS: Fifty consecutive rhinoplasty patients requiring bilateral osteotomies (100 total lateral osteotomies) were prospectively randomized (institutional review board no. 1341-1M) so that each patient received an external skin puncture perforating osteotomy on one side and an internal lateral osteotomy on the control side. The percutaneous skin puncture was made with the flat edge of the sharpened 2-mm unguarded osteotome. The puncture site was approximated with Steri-Strips as part of the postoperative nasal splint. All patients were evaluated for scarring on the left versus the right side of the face at 1, 3, and 6 weeks; 6 months; 1 year; and when possible up to 2 years after the operation. The median follow-up in this series is over 1 year. RESULTS: The external percutaneous approach caused a visible scar in three patients (6 percent) as noted by three blinded examiners. One of these patients (2 percent) required a scar revision. Care must be taken to cleanse the chisel before percutaneous entry. This practice should avoid a traumatic tattoo. CONCLUSION: Percutaneous osteotomies generally produce an imperceptible scar (94 percent) but may cause a visible scar in a small percentage of patients. PMID- 16267447 TI - Calibrated lateral crus stabilizer. AB - BACKGROUND: Fundamental shortcomings of the currently available instrumentation prompted the need for a device that facilitates stabilization of the lateral crus while at the same time allowing for precise measurement of the retained cartilage fragment along the anatomic axis of the lateral crus. METHODS: This article presents a new instrument, the calibrated lateral crus stabilizer, and highlights its unique features, which facilitate the precise and symmetric excision of the lateral crural cartilage. RESULTS: A lateral crus stabilizer with a calibration mechanism is introduced that provides the means for precise removal of the excess portion of the lower lateral cartilage during rhinoplasty. CONCLUSIONS: There was a need for improved instrumentation to stabilize the alar cartilage than what is currently available. New instrumentation is presented that answers that need. PMID- 16267448 TI - Assessing the long-term retention and permanency of acellular cross-linked porcine dermal collagen as a soft-tissue substitute. AB - BACKGROUND: Acellular porcine dermis has been used as a soft-tissue substitute in Europe since 1998 and in the United States since 2000. Processing renders the composite acellular, reducing antigenicity and crosslinking the dermal collagen. The purpose of this study was to determine the long-term histologic response of this material to the transfer into a subcutaneous soft-tissue location in an in vivo mouse model. METHODS: A total of 24 mice (ICR strain) underwent an initial procedure transplanting a segment of acellular porcine dermis into a dorsal subcutaneous pocket. The implants were examined for qualitative characteristics including rigidity, migration, inflammation, and healing response. Composites of skin, subcutaneous tissue, implant, and underlying muscle were harvested for histologic evaluation. The specimens were analyzed for inflammatory response, degree, and character of tissue incorporation and degree of implant resorption. RESULTS: One of the 24 animals studied experienced extrusion of the implant. The remaining 23 implants persisted within their respective hosts until the time the animals were killed. The volume and weight of all of the implants remained unchanged from the preoperative to postoperative measurements. Dystrophic calcification and bone formation was seen at 12 months. CONCLUSIONS: The results do not suggest characteristics optimal for implants used in soft-tissue augmentation for the purpose of aesthetics. The porcine dermis would appear from these results to be better suited for situations that require strength and permanence of the implant. PMID- 16267449 TI - Nonsurgical lower eyelid lift. PMID- 16267450 TI - Can you hear the bells? PMID- 16267451 TI - It's a matter of life. PMID- 16267452 TI - The three periorbital hollows: a paradigm for periorbital rejuvenation. PMID- 16267456 TI - Asian "double eyelid" blepharoplasty: the contribution of L. Fernandez. PMID- 16267457 TI - Harvesting fat from the infratemporal fossa. PMID- 16267459 TI - Thoracoepigastric flap in donor-site closure of the pectoralis major musculocutaneous transfer: axial or random pattern? PMID- 16267460 TI - Distally based dorsal forearm fasciosubcutaneous flap. PMID- 16267463 TI - Analysis of publications in three plastic surgery journals for the year 2002. PMID- 16267464 TI - Closure of large scalp defects by modified Gillies triple scalp flaps in patients with scalp tumors. PMID- 16267465 TI - Cerclage clamp. PMID- 16267466 TI - Skin ulceration in trigeminal trophic syndrome: report of a lesion occurring 22 years later. PMID- 16267467 TI - A new, easy, and pragmatic assessment of tear drainage after posttraumatic obstruction of the upper lacrimal canaliculus. PMID- 16267468 TI - Hypertrophy of the depressor septi nasi muscle. PMID- 16267469 TI - A composite cartilaginous graft. PMID- 16267470 TI - Pressure sore of the contralateral helical rim as a complication of middle ear surgery. PMID- 16267471 TI - A helpful adjunct to otoplasty surgery. PMID- 16267472 TI - Endoscopically assisted, intraorally approached palatoplasty. PMID- 16267474 TI - The "chewing gum test" for cleft palate speech. PMID- 16267473 TI - Palatal perforation as a result of neonatal sepsis. PMID- 16267475 TI - Is pseudocholinesterase elevation a risk for surgery? PMID- 16267476 TI - A safe way to insert breast implants. PMID- 16267477 TI - Barotrauma: an unrecognized mechanism for pneumothorax in breast augmentation. PMID- 16267478 TI - Implant found in thoracic cavity after breast augmentation. PMID- 16267479 TI - Removal of the difficult subcuticular suture. PMID- 16267480 TI - Use of aerosolized fibrin glue fixation after liposuction of the arms. PMID- 16267481 TI - Cross-digital vein graft in arterial repair. PMID- 16267483 TI - Does vacuum pressure extraction of fat affect the infranatant cellularity of liposuction specimens? PMID- 16267482 TI - The fish-skinning machine: an unusual source of hand trauma. PMID- 16267484 TI - Two scrotal calcinosis cases with different causal mechanisms. PMID- 16267485 TI - An unusual pressure sore site in a morbidly obese patient: infrapannicular fold. PMID- 16267486 TI - A simple bolstering method for optimizing skin graft take on the shaft of the penis. PMID- 16267487 TI - Pedicled flaps for foot defects in elderly patients: is duplex ultrasonography necessary? PMID- 16267489 TI - The perforator "plus" flap: a simple nomenclature for locoregional perforator based flaps. PMID- 16267488 TI - The congenital deficit of protein S as a new prognostic factor in microsurgery. PMID- 16267490 TI - What is the medicolegal implication of a Web-based three-dimensional interactive virtual reality plastic surgery package? PMID- 16267491 TI - Top ten reasons to hate the "V.A.C.". PMID- 16267494 TI - Ramelteon (Rozerem) for insomnia. PMID- 16267495 TI - Omega-3 polyunsaturated fatty acids (Omacor) for hypertriglyceridemia. PMID- 16267496 TI - Incidence of end-stage renal disease among persons with diabetes--United States, 1990-2002. AB - Diabetes mellitus is the leading cause of end-stage renal disease (ESRD) (i.e., kidney failure requiring dialysis or transplantation) in the United States, accounting for 44% of new cases of treated ESRD in 2002. To examine trends in ESRD attributed to diabetes mellitus (ESRD-DM) in the United States, CDC analyzed 1990-2002 data from the United States Renal Data System (USRDS) and the National Health Interview Survey (NHIS). This report summarizes the findings of that analysis, which indicated that, although the number of new cases of ESRD-DM increased overall, the incidence of ESRD-DM among persons with diabetes is not increasing among blacks, Hispanics, men, and persons aged 65-74 years, and is declining among persons aged <65 years, women, and whites. Continued interventions to reduce the prevalence of risk factors for kidney disease and improve diabetes care are needed to sustain and improve these trends. PMID- 16267497 TI - Global Measles and Rubella Laboratory Network, January 2004-June 2005. AB - Measles continues to be a leading cause of childhood morbidity and mortality in developing countries and an outbreak threat in the majority of countries. In 2000, measles was considered the fifth leading cause of childhood mortality, and the World Health Organization (WHO) estimated that approximately 777,000 measles associated deaths occurred worldwide. In 2001, WHO and the United Nations Children's Fund (UNICEF) developed a 5-year strategic plan, endorsed by the World Health Assembly in 2003, to reduce measles mortality by 50% by 2005 (relative to 1999 estimates) and to achieve and maintain interruption of indigenous measles transmission in large geographic areas with established measles elimination goals. This plan included strengthening routine vaccination coverage, providing a second opportunity for measles immunization to children, improving measles case management, and improving surveillance with laboratory confirmation of suspected measles cases. To date, four of six WHO regions have established measles elimination targets: the Americas Region (AMR) by 2000, the European Region (EUR) by 2010, the Eastern Mediterranean Region (EMR) by 2010, and the Western Pacific Region (WPR) by 2012. The remaining two WHO regions, the African (AFR) and South East Asian (SEAR) regions, are continuing work toward the measles mortality reduction goal. Likewise, to reduce the burden of disease from congenital rubella syndrome (CRS), currently estimated at 100,000 cases per year worldwide, several countries have developed or continue to develop rubella control programs, and AMR and EUR have established regional rubella elimination and CRS reduction goals, respectively. Because improved global surveillance is essential for monitoring progress toward mortality reduction and elimination of these diseases, WHO established the Measles and Rubella Laboratory Network (LabNet) in 2003 to promote case identification and confirmation. This report provides an update on the development of LabNet during January 2004-June 2005 and describes the geographic distribution of measles and rubella virus genotypes as of June 2005. PMID- 16267498 TI - Update: Ralstonia species associated with Vapotherm oxygen delivery devices- United States, 2005. AB - This report updates information on Ralstonia species associated with Vapotherm oxygen delivery devices (Vapotherm Inc., Stevensville, Maryland). CDC has obtained new information from a test developed by CDC and performed by The Children's Hospital of Philadelphia (Pennsylvania) to assess the efficacy of the new chlorine dioxide disinfection protocol recommended by Vapotherm. Although limited, this information suggests that the new protocol for disinfecting Vapotherm devices and cartridges might not achieve sustained bacterial control in certain situations. At this time, the optimal protocol to disinfect machines and cartridges that might contain very heavy biofilms is not known. PMID- 16267499 TI - Controlling tuberculosis in the United States. Recommendations from the American Thoracic Society, CDC, and the Infectious Diseases Society of America. AB - During 1993-2003, incidence of tuberculosis (TB) in the United States decreased 44% and is now occurring at a historic low level (14,874 cases in 2003). The Advisory Council for the Elimination of Tuberculosis has called for a renewed commitment to eliminating TB in the United States, and the Institute of Medicine has published a detailed plan for achieving that goal. In this statement, the American Thoracic Society (ATS), CDC, and the Infectious Diseases Society of America (IDSA) propose recommendations to improve the control and prevention of TB in the United States and to progress toward its elimination. This statement is one in a series issued periodically by the sponsoring organizations to guide the diagnosis, treatment, control, and prevention of TB. This statement supersedes the previous statement by ATS and CDC, which was also supported by IDSA and the American Academy of Pediatrics (AAP). This statement was drafted, after an evidence-based review of the subject, by a panel of representatives of the three sponsoring organizations. AAP, the National Tuberculosis Controllers Association, and the Canadian Thoracic Society were also represented on the panel. This statement integrates recent scientific advances with current epidemiologic data, other recent guidelines from this series, and other sources into a coherent and practical approach to the control of TB in the United States. Although drafted to apply to TB control activities in the United States, this statement might be of use in other countries in which persons with TB generally have access to medical and public health services and resources necessary to make a precise diagnosis of the disease; achieve curative medical treatment; and otherwise provide substantial science-based protection of the population against TB. This statement is aimed at all persons who advocate, plan, and work at controlling and preventing TB in the United States, including persons who formulate public health policy and make decisions about allocation of resources for disease control and health maintenance and directors and staff members of state, county, and local public health agencies throughout the United States charged with control of TB. The audience also includes the full range of medical practitioners, organizations, and institutions involved in the health care of persons in the United States who are at risk for TB. PMID- 16267500 TI - Genetic background of HSH in three Polish families and a patient with an X;9 translocation. AB - Hypomagnesemia with secondary hypocalcemia (HSH) is a rare inherited disease, characterised by neurological symptoms, such as tetany, muscle spasms and seizures, due to hypocalcemia. It has been suggested that HSH is genetically heterogeneous, but only one causative gene, TRPM6, on chromosome 9 has so far been isolated. We have now studied the genetic background of HSH in four Polish patients belonging to three families, and a HSH patient carrying an apparently balanced X;9 translocation. The translocation patient has long been considered as an example of the X-linked form of HSH. We identified six TRPM6 gene mutations, of which five were novel, in the Polish patients. All the alterations were either nonsense/splicing or missense mutations. The clinical picture of the patients was similar to the HSH patients reported earlier. No genotype-phenotype correlation could be detected. Sequencing did not reveal any TRPM6 or TRPM7 gene mutations in the female HSH patient with an X;9 translocation. Isolation of the translocation breakpoint showed that the chromosome 9 specific breakpoint mapped within satellite III repeat sequence. The X-chromosomal breakpoint was localised to the first intron of the vascular endothelial growth factor gene, VEGFD. No other sequence alterations were observed within the VEGFD gene. Even though the VEGFD gene was interrupted by the X;9 translocation, it seems unlikely that VEGFD is causing the translocation patient's HSH-like phenotype. Furthermore, re evaluation of patient's clinical symptoms suggests that she did not have a typical HSH. PMID- 16267501 TI - Genetics of the Lp(a)/apo(a) system in an autochthonous Black African population from the Gabon. AB - Plasma lipoprotein(a) (Lp(a)) is a quantitative trait associated with atherothrombotic disease in European and Asian populations. Lp(a) concentrations vary widely within and between populations, with Africans exhibiting on average two- to threefold higher Lp(a) levels and a different distribution compared to Europeans. The apo(a) gene locus on chromosome 6q26-27 (LPA, MIM 152200) has been identified as the major quantitative trait locus (QTL) for Lp(a) concentrations in Europeans and populations of African descent (North American and South African Blacks) but data on autochthonous Black Africans are lacking.Here, we have analysed Lp(a) plasma concentrations, apo(a) isoforms in plasma and four polymorphisms in the LPA gene in 31 African families with 54 children from Gabon. Weighted midparent-offspring regression estimated a heritability h2=0.76. The correlation of Lp(a) levels associated with LPA alleles identical by descent (IBD) resulted in a heritability estimate of 0.801. Our data demonstrate that Lp(a) concentrations are highly heritable in a Central African population without admixture and high Lp(a) (median 43 mg/dl). LPA is the major QTL, explaining most or all of the heritability of Lp(a) in this population. PMID- 16267502 TI - Carrier testing in minors: a systematic review of guidelines and position papers. AB - The objective of this article is to review all published normative ethical and clinical guidelines concerning the genetic carrier testing of minors. The databases Medline, Philosopher's Index, Biological Abstracts, Web of Science, and Google Scholar were searched using keywords relating to the carrier testing of children. We also searched the websites of the national bioethics committees indexed on the websites of WHO and the German Reference Center for Ethics in the Life Sciences, the Human Genetics Societies of various nations indexed on the website of the International Federation of Human Genetics Societies and related links, and the national medical associations indexed on the website of the World Medical Association. We retrieved 14 guidelines emanating from 24 different groups. All guidelines advanced the following preferences: (1) carrier testing should not be performed in children, and (2) testing should be deferred until the child can give proper informed consent to be tested. The guidelines varied in three areas: (a) the role of genetic services in ensuring that children are informed about their carrier status and associated risks when they are older; (b) exceptions to the general rule of withholding or deferring carrier testing; and (c) the communication of incidentally discovered carrier status. In the absence of compelling reasons, carrier testing of a child can reasonably be deferred until the child has the intellectual capacity needed to discern if and when to be tested. PMID- 16267503 TI - Intake of milk fat, reflected in adipose tissue fatty acids and risk of myocardial infarction: a case-control study. AB - OBJECTIVE: To study the association between content of fatty acids from milk fat (14:0, 15:0 and 17:0) in adipose tissue and risk of a first myocardial infarction (MI). DESIGN AND SUBJECTS: A case-control study with 99 patients and 98 population controls both men and postmenopausal women, age 45-75 year. Adipose tissue fatty acids were determined by gas-liquid chromatography. RESULTS: The content of 14:0, 14:1, 15:0, 17:0 and 17:1 were all significantly higher in adipose tissue of controls than of the patients. Age and sex adjusted odds ratios (OR) for MI were significantly reduced with increasing quartiles of 14:0, 14:1, 15:0 and 17:1 in adipose tissue, but except for 15:0 (OR = 0.36, 95% CI 0.13 0.99), the trend was no longer significant after further adjustment for waist-to hip ratio, smoking and family history for coronary heart disease. Correlations between 14:0 and 15:0 in adipose tissue, and waist-to-hip ratio were significantly negative (r = -0.22 for both, P < 0.01). CONCLUSION: Our study suggests that intake of dairy fat or some other component of dairy products, as reflected by C15:0 as marker in adipose tissue, may protect persons at increased risk from having a first MI, and that the causal effects may rely on other factors than serum cholesterol. SPONSORSHIP: Throne Holst's foundation for Nutrition Research, Research Council of Norway, The Norwegian Association of Margarine Producers, DeNoFa Fabriker A/S, TINE BA. PMID- 16267504 TI - Ecology: is speciation driven by species diversity? AB - Emerson and Kolm show that the proportion of species endemic to an island is positively related to its species richness and, assuming that endemism indexes speciation rate, they infer that greater species diversity accelerates diversification. Here we demonstrate that the same correlation between species richness and percentage endemism can arise even if within-island speciation is negligible, particularly when both endemism and species richness depend on attributes of islands (such as area) that influence the average age of resident populations. Island biogeography theory indicates that, where the average time to extinction is relatively long, diversity increases through colonization, irrespective of whether new species are formed; at the same time, islands on which populations persist for longer accumulate more endemic species as local populations differentiate and populations on neighbouring islands become extinct. We therefore suggest that species richness and endemism are correlated fortuitously owing to their mutual dependence on the life spans of populations on islands, which is unrelated to speciation itself. PMID- 16267509 TI - Turkey's evolution. PMID- 16267510 TI - Taking a stand on animal-rights violence. PMID- 16267511 TI - Clamp down on copycats. PMID- 16267513 TI - Drug firms donate compounds for anti-HIV gel. PMID- 16267514 TI - Wartime tactic doubles power of scarce bird-flu drug. PMID- 16267516 TI - Turkish rectors rally in support of university head thrown in jail. PMID- 16267515 TI - Universities scramble to assess scope of falsified results. PMID- 16267517 TI - Protists push animals aside in rule revamp. PMID- 16267519 TI - Floods fail to save canyon beaches. PMID- 16267520 TI - Expert witness: the scientists who testified against intelligent design. Interview by Geoff Brumfiel. PMID- 16267521 TI - Gene study raises fears for three-parent babies. PMID- 16267523 TI - Evolutionary theory: Personal effects. PMID- 16267524 TI - Star of the south. PMID- 16267525 TI - Meteorology: winds of change. PMID- 16267526 TI - Race is on for flu vaccine. PMID- 16267527 TI - Mapping the complexities of science and politics. PMID- 16267528 TI - Universal fungus register offers pattern for zoology. PMID- 16267534 TI - Wit and wisdom. PMID- 16267529 TI - UK must go on promoting and funding science. PMID- 16267535 TI - Intelligence: a gender bender. PMID- 16267536 TI - Astronomy: light on a dark place. PMID- 16267537 TI - Microbiology: algae and the vitamin mosaic. PMID- 16267539 TI - Global change: sea level and volcanoes. PMID- 16267540 TI - Structural biology: proteins flex to function. PMID- 16267541 TI - Cosmology: the infrared dawn of starlight. PMID- 16267543 TI - Chemical biology: bring them back alive. PMID- 16267544 TI - Obituary: Richard Doll (1912-2005). PMID- 16267545 TI - Theoretical mechanics: crowd synchrony on the Millennium Bridge. AB - Soon after the crowd streamed on to London's Millennium Bridge on the day it opened, the bridge started to sway from side to side: many pedestrians fell spontaneously into step with the bridge's vibrations, inadvertently amplifying them. Here we model this unexpected and now notorious phenomenon--which was not due to the bridge's innovative design as was first thought--by adapting ideas originally developed to describe the collective synchronization of biological oscillators such as neurons and fireflies. Our approach should help engineers to estimate the damping needed to stabilize other exceptionally crowded footbridges against synchronous lateral excitation by pedestrians. PMID- 16267546 TI - Nanoscale hydrodynamics: enhanced flow in carbon nanotubes. AB - Nanoscale structures that could mimic the selective transport and extraordinarily fast flow possible in biological cellular channels would have a wide range of potential applications. Here we show that liquid flow through a membrane composed of an array of aligned carbon nanotubes is four to five orders of magnitude faster than would be predicted from conventional fluid-flow theory. This high fluid velocity results from an almost frictionless interface at the carbon nanotube wall. PMID- 16267548 TI - A size of approximately 1 au for the radio source Sgr A* at the centre of the Milky Way. AB - Although it is widely accepted that most galaxies have supermassive black holes at their centres, concrete proof has proved elusive. Sagittarius A* (Sgr A*), an extremely compact radio source at the centre of our Galaxy, is the best candidate for proof, because it is the closest. Previous very-long-baseline interferometry observations (at 7 mm wavelength) reported that Sgr A* is approximately 2 astronomical units (au) in size, but this is still larger than the 'shadow' (a remarkably dim inner region encircled by a bright ring) that should arise from general relativistic effects near the event horizon of the black hole. Moreover, the measured size is wavelength dependent. Here we report a radio image of Sgr A* at a wavelength of 3.5 mm, demonstrating that its size is approximately 1 au. When combined with the lower limit on its mass, the lower limit on the mass density is 6.5 x 10(21)M(o) pc(-3) (where M(o) is the solar mass), which provides strong evidence that Sgr A* is a supermassive black hole. The power-law relationship between wavelength and intrinsic size (size proportional, variantwavelength(1.09)) explicitly rules out explanations other than those emission models with stratified structure, which predict a smaller emitting region observed at a shorter radio wavelength. PMID- 16267547 TI - Tracing the first stars with fluctuations of the cosmic infrared background. AB - The deepest space- and ground-based observations find metal-enriched galaxies at cosmic times when the Universe was less than 1 Gyr old. These stellar populations had to be preceded by the metal-free first stars, known as 'population III'. Recent cosmic microwave background polarization measurements indicate that stars started forming early--when the Universe was < or =200 Myr old. It is now thought that population III stars were significantly more massive than the present metal rich stellar populations. Although such sources will not be individually detectable by existing or planned telescopes, they would have produced significant cosmic infrared background radiation in the near-infrared, whose fluctuations reflect the conditions in the primordial density field. Here we report a measurement of diffuse flux fluctuations after removing foreground stars and galaxies. The anisotropies exceed the instrument noise and the more local foregrounds; they can be attributed to emission from population III stars, at an era dominated by these objects. PMID- 16267549 TI - Active control of slow light on a chip with photonic crystal waveguides. AB - It is known that light can be slowed down in dispersive materials near resonances. Dramatic reduction of the light group velocity-and even bringing light pulses to a complete halt-has been demonstrated recently in various atomic and solid state systems, where the material absorption is cancelled via quantum optical coherent effects. Exploitation of slow light phenomena has potential for applications ranging from all-optical storage to all-optical switching. Existing schemes, however, are restricted to the narrow frequency range of the material resonance, which limits the operation frequency, maximum data rate and storage capacity. Moreover, the implementation of external lasers, low pressures and/or low temperatures prevents miniaturization and hinders practical applications. Here we experimentally demonstrate an over 300-fold reduction of the group velocity on a silicon chip via an ultra-compact photonic integrated circuit using low-loss silicon photonic crystal waveguides that can support an optical mode with a submicrometre cross-section. In addition, we show fast (approximately 100 ns) and efficient (2 mW electric power) active control of the group velocity by localized heating of the photonic crystal waveguide with an integrated micro heater. PMID- 16267550 TI - Simulating micrometre-scale crystal growth from solution. AB - Understanding crystal growth is essential for controlling the crystallization used in industrial separation and purification processes. Because solids interact through their surfaces, crystal shape can influence both chemical and physical properties. The thermodynamic morphology can readily be predicted, but most particle shapes are actually controlled by the kinetics of the atomic growth processes through which assembly occurs. Here we study the urea-solvent interface at the nanometre scale and report kinetic Monte Carlo simulations of the micrometre-scale three-dimensional growth of urea crystals. These simulations accurately reproduce experimentally observed crystal growth. Unlike previous models of crystal growth, no assumption is made that the morphology can be constructed from the results for independently growing surfaces or from an a priori specification of surface defect concentration. This approach offers insights into the role of the solvent, the degree of supersaturation, and the contribution that extended defects (such as screw dislocations) make to crystal growth. It also connects observations made at the nanometre scale, through in situ atomic force microscopy, with those made at the macroscopic level. If extended to include additives, the technique could lead to the computer-aided design of crystals. PMID- 16267551 TI - Significant decadal-scale impact of volcanic eruptions on sea level and ocean heat content. AB - Ocean thermal expansion contributes significantly to sea-level variability and rise. However, observed decadal variability in ocean heat content and sea level has not been reproduced well in climate models. Aerosols injected into the stratosphere during volcanic eruptions scatter incoming solar radiation, and cause a rapid cooling of the atmosphere and a reduction in rainfall, as well as other changes in the climate system. Here we use observations of ocean heat content and a set of climate simulations to show that large volcanic eruptions result in rapid reductions in ocean heat content and global mean sea level. For the Mt Pinatubo eruption, we estimate a reduction in ocean heat content of about 3 x 10(22) J and a global sea-level fall of about 5 mm. Over the three years following such an eruption, we estimate a decrease in evaporation of up to 0.1 mm d(-1), comparable to observed changes in mean land precipitation. The recovery of sea level following the Mt Pinatubo eruption in 1991 explains about half of the difference between the long-term rate of sea-level rise of 1.8 mm yr(-1) (for 1950-2000), and the higher rate estimated for the more recent period where satellite altimeter data are available (1993-2000). PMID- 16267552 TI - Crustal rheology of the Himalaya and Southern Tibet inferred from magnetotelluric data. AB - The Cenozoic collision between the Indian and Asian continents formed the Tibetan plateau, beginning about 70 million years ago. Since this time, at least 1,400 km of convergence has been accommodated by a combination of underthrusting of Indian and Asian lithosphere, crustal shortening, horizontal extrusion and lithospheric delamination. Rocks exposed in the Himalaya show evidence of crustal melting and are thought to have been exhumed by rapid erosion and climatically forced crustal flow. Magnetotelluric data can be used to image subsurface electrical resistivity, a parameter sensitive to the presence of interconnected fluids in the host rock matrix, even at low volume fractions. Here we present magnetotelluric data from the Tibetan-Himalayan orogen from 77 degrees E to 92 degrees E, which show that low resistivity, interpreted as a partially molten layer, is present along at least 1,000 km of the southern margin of the Tibetan plateau. The inferred low viscosity of this layer is consistent with the development of climatically forced crustal flow in Southern Tibet. PMID- 16267553 TI - Proteorhodopsin in the ubiquitous marine bacterium SAR11. AB - Proteorhodopsins are light-dependent proton pumps that are predicted to have an important role in the ecology of the oceans by supplying energy for microbial metabolism. Proteorhodopsin genes were first discovered through the cloning and sequencing of large genomic DNA fragments from seawater. They were later shown to be widely distributed, phylogenetically diverse, and active in the oceans. Proteorhodopsin genes have not been found in cultured bacteria, and on the basis of environmental sequence data, it has not yet been possible to reconstruct the genomes of uncultured bacterial strains that have proteorhodopsin genes. Although the metabolic effect of proteorhodopsins is uncertain, they are thought to function in cells for which the primary mode of metabolism is the heterotrophic assimilation of dissolved organic carbon. Here we report that SAR11 strain HTCC1062 ('Pelagibacter ubique'), the first cultivated member of the extraordinarily abundant SAR11 clade, expresses a proteorhodopsin gene when cultured in autoclaved seawater and in its natural environment, the ocean. The Pelagibacter proteorhodopsin functions as a light-dependent proton pump. The gene is expressed by cells grown in either diurnal light or in darkness, and there is no difference between the growth rates or cell yields of cultures grown in light or darkness. PMID- 16267554 TI - Algae acquire vitamin B12 through a symbiotic relationship with bacteria. AB - Vitamin B12 (cobalamin) was identified nearly 80 years ago as the anti-pernicious anaemia factor in liver, and its importance in human health and disease has resulted in much work on its uptake, cellular transport and utilization. Plants do not contain cobalamin because they have no cobalamin-dependent enzymes. Deficiencies are therefore common in strict vegetarians, and in the elderly, who are susceptible to an autoimmune disorder that prevents its efficient uptake. In contrast, many algae are rich in vitamin B12, with some species, such as Porphyra yezoensis (Nori), containing as much cobalamin as liver. Despite this, the role of the cofactor in algal metabolism remains unknown, as does the source of the vitamin for these organisms. A survey of 326 algal species revealed that 171 species require exogenous vitamin B12 for growth, implying that more than half of the algal kingdom are cobalamin auxotrophs. Here we show that the role of vitamin B12 in algal metabolism is primarily as a cofactor for vitamin B12-dependent methionine synthase, and that cobalamin auxotrophy has arisen numerous times throughout evolution, probably owing to the loss of the vitamin B12-independent form of the enzyme. The source of cobalamin seems to be bacteria, indicating an important and unsuspected symbiosis. PMID- 16267555 TI - The transcription factor Engrailed-2 guides retinal axons. AB - Engrailed-2 (En-2), a homeodomain transcription factor, is expressed in a caudal to-rostral gradient in the developing midbrain, where it has an instructive role in patterning the optic tectum--the target of topographic retinal input. In addition to its well-known role in regulating gene expression through its DNA binding domain, En-2 may also have a role in cell-cell communication, as suggested by the presence of other domains involved in nuclear export, secretion and internalization. Consistent with this possibility, here we report that an external gradient of En-2 protein strongly repels growth cones of Xenopus axons originating from the temporal retina and, conversely, attracts nasal axons. Fluorescently tagged En-2 accumulates inside growth cones within minutes of exposure, and a mutant form of the protein that cannot enter cells fails to elicit axon turning. Once internalized, En-2 stimulates the rapid phosphorylation of proteins involved in translation initiation and triggers the local synthesis of new proteins. Furthermore, the turning responses of both nasal and temporal growth cones in the presence of En-2 are blocked by inhibitors of protein synthesis. The differential guidance of nasal and temporal axons reported here suggests that En-2 may participate directly in topographic map formation in the vertebrate visual system. PMID- 16267556 TI - A protein interaction network of the malaria parasite Plasmodium falciparum. AB - Plasmodium falciparum causes the most severe form of malaria and kills up to 2.7 million people annually. Despite the global importance of P. falciparum, the vast majority of its proteins have not been characterized experimentally. Here we identify P. falciparum protein-protein interactions using a high-throughput version of the yeast two-hybrid assay that circumvents the difficulties in expressing P. falciparum proteins in Saccharomyces cerevisiae. From more than 32,000 yeast two-hybrid screens with P. falciparum protein fragments, we identified 2,846 unique interactions, most of which include at least one previously uncharacterized protein. Informatic analyses of network connectivity, coexpression of the genes encoding interacting fragments, and enrichment of specific protein domains or Gene Ontology annotations were used to identify groups of interacting proteins, including one implicated in chromatin modification, transcription, messenger RNA stability and ubiquitination, and another implicated in the invasion of host cells. These data constitute the first extensive description of the protein interaction network for this important human pathogen. PMID- 16267557 TI - The Plasmodium protein network diverges from those of other eukaryotes. AB - Plasmodium falciparum is the pathogen responsible for over 90% of human deaths from malaria. Therefore, it has been the focus of a considerable research initiative, involving the complete DNA sequencing of the genome, large-scale expression analyses, and protein characterization of its life-cycle stages. The Plasmodium genome sequence is relatively distant from those of most other eukaryotes, with more than 60% of the 5,334 encoded proteins lacking any notable sequence similarity to other organisms. To systematically elucidate functional relationships among these proteins, a large two-hybrid study has recently mapped a network of 2,846 interactions involving 1,312 proteins within Plasmodium. This network adds to a growing collection of available interaction maps for a number of different organisms, and raises questions about whether the divergence of Plasmodium at the sequence level is reflected in the configuration of its protein network. Here we examine the degree of conservation between the Plasmodium protein network and those of model organisms. Although we find 29 highly connected protein complexes specific to the network of the pathogen, we find very little conservation with complexes observed in other organisms (three in yeast, none in the others). Overall, the patterns of protein interaction in Plasmodium, like its genome sequence, set it apart from other species. PMID- 16267558 TI - A putative stimulatory role for activator turnover in gene expression. AB - The ubiquitin-proteasome system (UPS) promotes the destruction of target proteins by attaching to them a ubiquitin chain that is recognized by the 26S proteasome. The UPS influences most cellular processes, and its targets include transcriptional activators that are primary determinants of gene expression. Emerging evidence indicates that non-proteolytic functions of the UPS might stimulate transcriptional activity. Here we show that the proteolysis of some transcriptional activators by the UPS can stimulate their function. We focused on the role of UPS-dependent proteolysis in the function of inducible transcriptional activators in yeast, and found that inhibition of the proteasome reduced transcription of the targets of the activators Gcn4, Gal4 and Ino2/4. In addition, mutations in SCF(Cdc4), the ubiquitin ligase for Gcn4 (ref. 5), or mutations in ubiquitin that prevent degradation, also impaired the transcription of Gcn4 targets. These transcriptional defects were manifested despite the enhanced abundance of Gcn4 on cognate promoters. Proteasome inhibition also decreased the association of RNA polymerase II with Gcn4, Gal4 and Ino2/4 targets, as did mutations in SCF(Cdc4) for Gcn4 targets. Expression of a stable phospho-site mutant of Gcn4 (ref. 7) or disruption of the kinases that target Gcn4 for turnover alleviated the sensitivity of Gcn4 activity to defects in the UPS. PMID- 16267559 TI - Intrinsic dynamics of an enzyme underlies catalysis. AB - A unique feature of chemical catalysis mediated by enzymes is that the catalytically reactive atoms are embedded within a folded protein. Although current understanding of enzyme function has been focused on the chemical reactions and static three-dimensional structures, the dynamic nature of proteins has been proposed to have a function in catalysis. The concept of conformational substates has been described; however, the challenge is to unravel the intimate linkage between protein flexibility and enzymatic function. Here we show that the intrinsic plasticity of the protein is a key characteristic of catalysis. The dynamics of the prolyl cis-trans isomerase cyclophilin A (CypA) in its substrate free state and during catalysis were characterized with NMR relaxation experiments. The characteristic enzyme motions detected during catalysis are already present in the free enzyme with frequencies corresponding to the catalytic turnover rates. This correlation suggests that the protein motions necessary for catalysis are an intrinsic property of the enzyme and may even limit the overall turnover rate. Motion is localized not only to the active site but also to a wider dynamic network. Whereas coupled networks in proteins have been proposed previously, we experimentally measured the collective nature of motions with the use of mutant forms of CypA. We propose that the pre-existence of collective dynamics in enzymes before catalysis is a common feature of biocatalysts and that proteins have evolved under synergistic pressure between structure and dynamics. PMID- 16267563 TI - Hypertension labelling was associated with poorer self-rated health in the Third US National Health and Nutrition Examination Survey. AB - Individuals labelled as having hypertension tend to report poor self-rated health (SRH), but it is unclear whether this association is independent of actual hypertension, socioeconomic status and adiposity, and extends across racial and ethnic groups. In a cross-sectional study we compared hypertensive and normotensive individuals (N = 19,057) who varied in whether they had ever been labelled hypertensive. Blood pressure was measured in participants' homes and mobile examination centres in the United States as part of the Third National Health and Nutrition Examination Survey, 1988-1994. The main outcome measure was global SRH. Hypertensive labelling was associated with poorer SRH and was independent of established SRH predictors, antihypertensive medication use, body mass index, and hypertension status (adjusted odds ratio (OR) = 1.79, 95% confidence interval (CI), 1.61-1.99). Hypertension was also associated with poorer SRH (OR = 1.26; 95% CI 1.09-1.46) but this association was eliminated by adjustment for hypertensive labelling (OR 1.06; 95% CI 0.92-1.22). These effects were consistent across non-Hispanic white, non-Hispanic black, and Hispanic subgroups. Individuals labelled hypertensive are more likely to have lower SRH and this labelling effect predominates over that of actual hypertension. Public health efforts to increase the number of individuals screened for high blood pressure may successfully detect the presence of hypertension but may also reduce health-related quality of life as measured by global SRH. PMID- 16267564 TI - A haplotype in the human Sur2 gene is associated with essential hypertension. PMID- 16267565 TI - Inventories for male and female sexual dysfunctions. AB - Several illnesses can distress sexual health and disrupt sexuality, an integral part of being human. These illnesses are not different in origin from those affecting other health areas, but, because sexuality is involved, effective communication and empathy between the physician and the patient may be severely hindered by negative feelings such as anxiety and guilt. A detailed general and sexual history is, however, an essential step in evaluating patients for sexual dysfunction (SD). Finding the correct way to ask questions and to decode answers on sexual health and disease might be difficult and, in some way, embarrassing. Hence, validated and standardized sexual inventories might help physicians confront SD. These case-history tools have the advantage of being standardized, easy to administer and score, relatively unobtrusive and substantially inexpensive. This review describes the main sexual inventories hitherto described and validated in different sexual areas of health and disease, and the advantages of the two main formats available to clinicians, that is, structured interviews (SIs) and self-report questionnaires (SRQs). Both types of inventories are composed of a set of standardized, written probe questions requiring a finite number of responses, driven by an interviewer (SIs) or by the patients themselves (SRQs). SRQs allow more time and intimacy to organize and develop answers to delicate questions, as are those on sexual life. In addition, SRQs could also be very useful in quantifying disease severity and treatment outcome. On the other hand, SIs help achieve a better patient-physician relationship and reduce the risk of misunderstandings. PMID- 16267566 TI - Immune response to lentiviral bilirubin UDP-glucuronosyltransferase gene transfer in fetal and neonatal rats. AB - Gene therapy for inherited disorders might cause an immune response to the therapeutic protein. A solution would be to introduce the gene in the fetal or neonatal period, which should lead to tolerization. Lentiviral vectors mediate long-term gene expression, and are well suited for gene therapy early in development. A model for fetal or neonatal gene therapy is the inherited disorder of bilirubin metabolism, Crigler-Najjar disease (CN). The absence of bilirubin UDP-glucoronyltransferase (UGT1A1) activity in CN patients causes high serum levels of unconjugated bilirubin and brain damage in infancy. CN is attractive for the development of gene therapy because the mutant Gunn rat closely mimics the human disease. Injection of UGT1A1 lentiviral vectors corrected the hyperbilirubinemia for more than a year in rats injected as fetuses and for up to 18 weeks in rats injected the day of birth. UGT1A1 gene transfer was confirmed by the presence of bilirubin glucuronides in bile. All animals injected with UGT1A1 lentiviral vectors developed antibodies to UGT1A1. Animals injected with green fluorescent protein (GFP) lentiviral vectors did not develop antibodies to GFP. Our results indicate that fetal and neonatal gene therapy with immunogenic proteins such as UGT1A1 does not necessarily lead to tolerization. PMID- 16267567 TI - Reconstituted influenza virus envelopes as an efficient carrier system for cellular delivery of small-interfering RNAs. AB - Application of RNA interference for in vivo evaluation of gene function or for therapeutic interventions has been hampered by a lack of suitable delivery methods for small interfering RNA (siRNA). Here, we present reconstituted viral envelopes (virosomes) derived from influenza virus as suitable vehicles for in vitro as well as in vivo delivery of siRNAs. Virosomes are vesicles that bear in their membrane the influenza virus spike protein hemagglutinin (HA). This protein mediates binding of native virus to and fusion with cellular target membranes. Accordingly, virosomes with membrane-incorporated HA bind to cells, are taken up by receptor-mediated endocytosis, and fuse with the endosomal membrane to release their contents into the cytoplasm. When complexed to cationic lipids, siRNA was successfully encapsulated in virosomes. Virosomes with encapsulated siRNA fused with target membranes in a pH-dependent manner and delivered the encapsulated siRNA to several cell lines in vitro. Virosome-delivered siRNA markedly downregulated the synthesis of newly induced and constitutively expressed green fluorescent protein. Moreover, intraperitoneal injection of siRNA-loaded virosomes resulted in delivery of the nucleotides to cells in the peritoneal cavity. Our results indicate that virosomes are a promising delivery device for in vivo application, especially where topical administration of siRNA, for example, to the respiratory tract is envisaged. PMID- 16267568 TI - Selective proapoptotic activity of a secreted recombinant antibody/AIF fusion protein in carcinomas overexpressing HER2. AB - Apoptosis-inducing factor (AIF) represents a caspase-independent apoptotic pathway in the cell, and a mitochondrial localization sequence-truncated AIF (AIFDelta1-120) can be relocated from the cytoplasm to the nucleus and exhibit a constitutive proapoptotic activity. Here, we generated a chimeric immuno-AIF protein, which comprised an HER2 antibody, a Pseudomonas exotoxin translocation domain and AIFDelta1-120. Human Jurkat cells transfected with the immuno-AIF gene could express and secrete the chimeric protein, which selectively recognized HER2 overexpressing tumor cells and was endocytosed. Subsequent cleavage of truncated AIF from immuno-AIF and its release from the internalized vesicles resulted in apoptosis of tumor cells. Intramuscular injection of the immuno-AIF gene caused significant suppression of tumors and substantially prolonged mice survival in an HER2-overexpressing xenograft tumor model. Our study demonstrates the feasibility of the immuno-AIF gene as a novel approach to treating cancers that overexpress HER2. PMID- 16267569 TI - A membrane antibody receptor for noninvasive imaging of gene expression. AB - Monitoring gene expression is important to optimize gene therapy protocols and ensure that the proper tissue distribution is achieved in clinical practice. We developed a noninvasive imaging system based on the expression of artificial antibody receptors to trap hapten-labeled imaging probes. Functional membrane bound anti-dansyl antibodies (DNS receptor) were stably expressed on melanoma cells in vitro and in vivo. A bivalent (DNS)2-diethylenetriaminepentaacetic 111Indium probe specifically bound to cells that expressed DNS receptors but not control scFv receptors. Importantly, the 111In probe preferentially localized to DNS receptors but not control receptors on tumors in mice as assessed by gamma camera imaging. By 48 h after intravenous injection, the uptake of the probe in tumors expressing DNS receptors was 72 times greater than the amount of probe in the blood. This targeting strategy may allow noninvasive assessment of the location, extent and persistence of gene expression in living animals and in the clinic. PMID- 16267570 TI - RNA knockdown as a potential therapeutic strategy in Parkinson's disease. AB - Parkinson's disease is a prevalent progressive degenerative disorder of the elderly. There is a current need for novel therapeutic strategies because the standard levodopa pharmacotherapy is only temporarily efficacious. Recently, there have been some high-profile successful preclinical results obtained in animal models of neurological disorders using small interfering RNAs delivered by viral vectors. RNA interference can theoretically be applied to Parkinson's disease since over-expression of various proteins is known to kill the dopamine neurons of the substantia nigra in animal models and in familial forms of Parkinson's disease. Potential RNA interfering strategies and caveats are discussed in this review. PMID- 16267571 TI - Truncated vesicular stomatitis virus G protein improves baculovirus transduction efficiency in vitro and in vivo. AB - Pseudotyping of viral vectors has been widely used to enhance viral transduction efficiency. One of the most popular pseudotyping proteins has been the G-protein of the vesicular stomatitis virus, VSV-G. In the present study, we show that the 21-amino-acid ectodomain with transmembrane and cytoplasmic tail domains of VSV-G (VSV-GED) augments baculovirus-mediated gene delivery in vertebrate cells by aiding viral entry. The VSV-GED pseudotyped virus replicated efficiently in insect cells yielding high titers. Five out of six studied cell lines showed improved transduction, as measured by a number of transduced cells or transgene expression level. Nearly 15-fold increase in the transduction efficiency was detected in rat malignant glioma cells as compared to the control virus. In the rat brain, transgene expression could be detected in the walls of lateral ventricles and in subarachnoid membranes. Increased transduction efficiency was also observed in the rabbit muscle. Our results suggest that VSV-GED enhances baculoviral gene transfer by augmenting gp64-mediated endosomal release. Moreover, no cytotoxicity was associated with improved gene transfer efficiency. Thus, VSV-GED pseudotyping provides a simple means to enhance baculovirus mediated gene transfer in vitro and in vivo. PMID- 16267574 TI - Identifying and counting point defects in carbon nanotubes. AB - The prevailing conception of carbon nanotubes and particularly single-walled carbon nanotubes (SWNTs) continues to be one of perfectly crystalline wires. Here, we demonstrate a selective electrochemical method that labels point defects and makes them easily visible for quantitative analysis. High-quality SWNTs are confirmed to contain one defect per 4 microm on average, with a distribution weighted towards areas of SWNT curvature. Although this defect density compares favourably to high-quality, silicon single-crystals, the presence of a single defect can have tremendous electronic effects in one-dimensional conductors such as SWNTs. We demonstrate a one-to-one correspondence between chemically active point defects and sites of local electronic sensitivity in SWNT circuits, confirming the expectation that individual defects may be critical to understanding and controlling variability, noise and chemical sensitivity in SWNT electronic devices. By varying the SWNT synthesis technique, we further show that the defect spacing can be varied over orders of magnitude. The ability to detect and analyse point defects, especially at very low concentrations, indicates the promise of this technique for quantitative process analysis, especially in nanoelectronics development. PMID- 16267573 TI - Anharmonic motions of Kr in the clathrate hydrate. AB - The anomalous glass-like thermal conductivity of crystalline clathrates has been suggested to be the result of the scattering of thermal phonons of the framework by 'rattling' motions of the guests in the clathrate cages. Using the site specific (83)Kr nuclear resonant inelastic scattering spectroscopy in combination with conventional incoherent inelastic neutron scattering and molecular-dynamics simulations, we provide unambiguous evidence and characterization of the effects on these guest-host interactions in a structure-II Kr clathrate hydrate. The resonant scattering of phonons led to unprecedented large anharmonic motions of the guest atoms. The anharmonic interaction underlies the anomalous thermal transport in this system. Clathrates are prototypical models for a class of crystalline framework materials with glass-like thermal conductivity. The explanation of the unusual molecular dynamics has a wide implication for the understanding of the thermal properties of disordered solids and structural glasses. PMID- 16267575 TI - Nanoparticle networks reduce the flammability of polymer nanocomposites. AB - Synthetic polymeric materials are rapidly replacing more traditional inorganic materials, such as metals, and natural polymeric materials, such as wood. As these synthetic materials are flammable, they require modifications to decrease their flammability through the addition of flame-retardant compounds. Environmental regulation has restricted the use of some halogenated flame retardant additives, initiating a search for alternative flame-retardant additives. Nanoparticle fillers are highly attractive for this purpose, because they can simultaneously improve both the physical and flammability properties of the polymer nanocomposite. We show that carbon nanotubes can surpass nanoclays as effective flame-retardant additives if they form a jammed network structure in the polymer matrix, such that the material as a whole behaves rheologically like a gel. We find this kind of network formation for a variety of highly extended carbon-based nanoparticles: single- and multiwalled nanotubes, as well as carbon nanofibres. PMID- 16267576 TI - DNA-programmed assembly of nanostructures. AB - DNA is a unique material for nanotechnology since it is possible to use base sequences to encode instructions for assembly in a predetermined fashion at the nanometre scale. Synthetic oligonucleotides are readily obtained by automated synthesis and numerous techniques have been developed for conjugating DNA with other materials. The exact spatial positioning of materials is crucial for the future development of complex nanodevices and the emerging field of DNA nanotechnology is now exploring DNA-programmed processes for the assembly of organic compounds, biomolecules, and inorganic materials. PMID- 16267577 TI - Probing dipeptide trans/cis stereochemistry using pH control of thiopeptide analogues, and application to the PepT1 transporter. AB - The stereochemistry of thiodipeptides of proline [e.g. Ala-Psi[CS-N]-Pro] can be controlled using pH, allowing the trans-preference for substrates of the peptide transporter PepT1 to be confirmed. PMID- 16267578 TI - Selective transannular ring transformations in azirino-fused eight-membered O,N- or S,N-heterocycles. AB - The first examples of transannular ring transformations in azirino-fused eight membered O,N- or S,N-heterocycles involving selective aziridine ring opening and medium-sized ring contraction are described, which provide an access to functionalized 1,4-benzox(thi)azines or 1,3-benzox(thi)azoles. PMID- 16267579 TI - Aminocatalytic preparation of bisindolylalkanes. AB - An aminocatalytic method for the formation of bisindolylalkanes is described; the reaction proceeds effectively at ambient temperature in the presence of moisture and air for both aldehyde and ketone substrates and has been applied to the one step preparation of a series of naturally-occurring bis- and tris-indolylalkanes. PMID- 16267580 TI - Mechanistic divergence of two closely related aldol-like enzyme-catalysed reactions. AB - The analysis of the interaction of threose 4-phosphate and 2-deoxyerythrose 4 phosphate with 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase (DAH7PS) reveals previously unrecognised mechanistic differences between the DAH7PS catalysed reaction and that catalysed by the closely related enzyme, 3-deoxy-D manno-octulosonate 8-phosphate synthase (KDO8PS). PMID- 16267581 TI - A novel azetidinyl gamma-lactam based peptide with a preference for beta-turn conformation. AB - Novel azetidinyl gamma-lactam based peptides have been synthesized with only compound showing a preference for the beta-turn conformation. PMID- 16267582 TI - Ring currents in the porphyrins: pi shielding, delocalisation pathways and the central cation. AB - It is shown that the ipsocentric orbital-based model explains how the charge of the central cation drives the delocalisation pathway in metalloporphyrins. A positive charge +Ze at the centre of the porphin ring gives rise to a two-way radial transfer of charge within the pi structure of the porphin macrocycle. This manifests itself in a change of pathway of the global pi current, as Z increases from Z = 0, from an inner- through a bifurcated- to an outer-pathway. Changes of pathway can be interpreted in terms of a specific pi shielding effect whereby electrons in high-lying pi orbitals are screened from the central charge by the electrons in lower-lying orbitals of the same symmetry. These changes in pi structure are essentially independent of accompanying changes in the sigma structure. PMID- 16267583 TI - Amino acid ester salt recognition by ferrocene-based ditopic receptor bearing oligoethylene glycol with pendant bipy subunits: CV, UV-vis and ESR studies. AB - The novel ferrocene-based ditopic receptor 1 was synthesized. This receptor bears two oligoethylene glycol arms with pendant 2,2'-bipyridine unit at the identical cyclopentadienyl rings, Cu(I) cation binds to 1 to form the 1 : 1 complex (1.Cu(I)) with the cavity consisting of polyether, and the resulting complex acts as a receptor for amino acid ester salts to give the ditopic complex (1.Cu(I).AAOMe-HCl). The 1H NMR spectrum of 1.Cu(I).LeuOMe-HCl exhibits strong broadening at the bipyridine region, and the ESR spectrum of the same sample gives the signals assigned as Cu(II) species. With these data, the binding of 1.Cu(I) towards AAOMe.HCl leads to the conformational change, and the Cu(I) complex is simultaneously oxidized to the Cu(II) complex. PMID- 16267585 TI - Control of rotor motion in a light-driven molecular motor: towards a molecular gearbox. AB - Controlled intramolecular movement and coupling of motor and rotor functions is exerted by this new molecular device. The rate of rotation of the rotor part of the molecule can be adjusted by alteration of the conformation of the motor part of the molecule. For all states of the motor part, different rates of rotation were measured for the rotor part. Conversion between the four propeller orientations was achieved by irradiation and heating. PMID- 16267584 TI - New C16 fatty-acid-based oxylipin pathway in the marine diatom Thalassiosira rotula. AB - An unprecedented series of C16 oxylipins (1-8) has been characterized from the marine diatom Thalassiosira rotula. Absolute stereochemistry of the major alcohols 1 and 3 was determined to be 9S by spectroscopic and chemical methods. All the described products are formally derived by unprecedented enzymatic oxidation of C16 fatty acids. Conversion of hexadeca-6,9,12-trienoic acid (C16:3 omega-4) into 3 unequivocally established the occurrence of (at least) a specific 9S-oxygenase activity. To the best of our knowledge, the present data reveal for the first time the existence of an organic network of oxygenase-mediated transformations that require C16 fatty acids as substrates in living cells. PMID- 16267586 TI - Synthesis and binding studies of 2-arylapomorphines. AB - From codeine, four different 2-aryl substituted apomorphines were synthesised in 6 steps each. Oxidation of codeine with IBX followed by acid catalysed rearrangement gave morphothebaine, which was selectively triflylated at the 2 position and subsequently O-acetylated at the 11-position. The resulting triflate was coupled in a Suzuki-Miyaura type reaction with a series of 4-substituted arylboronic esters which, after deprotection, gave the desired 2-aryl apomorphines. The analogues were tested for affinity towards a range of dopaminergic, serotonergic and adrenergic receptors. 2-(4-Hydroxyphenyl) apomorphine exhibited high affinity for the dopamine D2 receptor. A putative ligand-receptor interaction was put forward. PMID- 16267587 TI - Mechanistic studies of La3+ and Zn2+-catalyzed methanolysis of O-ethyl O-aryl methylphosphonate esters. An effective solvolytic method for the catalytic destruction of phosphonate CW simulants. AB - The kinetics of methanolysis of six O-ethyl O-aryl methylphosphonates (6a-f) promoted by methoxide, La3+ and 1,5,9-triazacyclododecane complex of Zn2+(-OCH3) (5:Zn2+(-OCH3)) were studied as simulants for chemical warfare (CW) agents, and analyzed through the use of Bronsted plots. The beta(lg) values are, respectively, -0.76, -1.26 and -1.06, pointing to significant weakening of the P OAr bond in the transition state. For the metal-catalyzed reactions the data are consistent with a concerted process where the P-OAr bond rupture has progressed to the extent of 84% in the La3+ reaction and ca. 70% in the Zn2+ catalyzed reaction. The catalysis afforded by the metal ions is remarkable, being about 10(6)-fold and 10(8)-fold for poor and good leaving groups, respectively, relative to the background reactions at pH 9.1. Solvent deuterium kinetic isotope studies for two of the substrates promoted by 5:Zn2+(-OCH3) give kH/kD = 1.0 +/- 0.1, consistent with a nucleophilic mechanism. A unified mechanism for the metal catalyzed reactions is presented which involves pre-equilibrium coordination of the substrate to the metal ion followed by intramolecular delivery of a coordinated methoxide. PMID- 16267588 TI - A new route to 2-C- and 4-C-branched sugars by palladium-indium bromide-mediated carbonyl allylation. AB - Palladium-catalysed carbonyl allylations can be effectively applied to the regio- and diastereoselective synthesis of 2-C- and 4-C-branched sugars from allylic esters or carbonates via the formation of pi-allylpalladium(II) intermediates and their reductive transmetalation with indium(I) bromide. PMID- 16267589 TI - Michael, Michael-aldol and Michael-Michael reactions of enolate equivalents of butane-2,3-diacetal protected glycolic acid derivatives. AB - Consecutive coupling reactions of butane-2,3-diacetal protected glycolic acid derivatives with Michael acceptors and aldehydes are reported. An enantiopure sample of this building block was used to kinetically resolve a chiral Michael acceptor present as a racemic mixture of enantiomers. PMID- 16267590 TI - Cyclic and acyclic sulfonimides in reactions with Rh(II)-ketocarbenoids: a new access to chemoselective O-functionalization of the imidic carbonyl groups. AB - Catalytic decomposition of diazoacetylacetone, diazoacetoacetic, diazomalonic, and diazoacetic esters using dirhodium tetraacetate in the presence of isothiazol 3(2H)-one 1,1-dioxides and a number of N-(arenesulfonyl)carboxamides in solutions of methylene chloride or dichloroethane gives rise to O-alkylation of the imidic carbonyl groups by Rh(II)-carbenoids and the formation of O-alkylimidates as the final products. The reaction proceeds with high chemoselectivity via carbonyl ylides and offers a powerful method for the synthesis in good yields of the imidates with polyfunctional O-alkyl groups. On the basis of X-ray analysis and 1H- and 13C-NMR studies it was shown that the resulting acyclic O-alkylimidates have the E-configuration in the solid state and in solution. Unlike acyclic analogues, the cyclic carbonyl ylide derived from substituted diazosaccharin by intramolecular cyclization of the appropriate diketocarbenoid is capable of reacting with DMAD in a 1,3-cycloaddition process. PMID- 16267591 TI - New routes to beta-cycloalkylalanine derivatives using serine-derived organozinc reagents. AB - Two distinct routes to beta-cycloalkylalanine derivatives have been developed. The first route employs the reaction of the iodoalanine-derived zinc-copper reagent 2 with cycloalk-1-en-3-yl phosphates, and the second uses the palladium catalysed coupling of the iodoalanine-derived zinc reagent 1 with cycloalkenyl triflates; in each case, catalytic hydrogenation of the unsaturated product leads to the protected beta-cycloalkylalanine. The latter route allows access to a range of cycloalkyl derivatives, with ring sizes of 5-8. beta-(1-Methyl-1 cyclohexyl)alanine may be prepared using reaction of the zinc-copper reagent 2 with 3-methyl-2-cyclohexenyl chloride, followed by hydrogenation. The corresponding cyclopentyl derivative may be prepared by reaction of the same zinc copper reagent 2 with diethyl geranylphosphate, followed by ring-closing metathesis and hydrogenation. PMID- 16267592 TI - A concise synthetic route to the conduritols from pentoses. AB - A short synthetic strategy for preparation of the conduritols is described. The key step employs a zinc-mediated fragmentation of protected methyl 5-deoxy-5-iodo d-pentofuranosides followed by an allylation of the intermediate aldehyde in the same pot. The allylation is performed with 3-bromopropenyl benzoate and occurs with good diastereoselectivity. An amino group can be introduced in the product by trapping the intermediate aldehyde as the imine prior to the allylation. The functionalised 1,7-octadienes, thus obtained, are converted into protected conduritols by ring-closing olefin metathesis. PMID- 16267593 TI - Mild and efficient method for the cleavage of benzylidene acetals by using erbium (III) triflate. AB - Er(OTf)3 is proposed as new efficient Lewis acid catalyst in a mild deprotection protocol of benzylidene derivatives. In a modified procedure, where acetic anhydride is used as the reaction solvent, the simultaneous cleavage of the benzylidene acetal and the peracetylation of the substrates is obtained in quantitative yields and very short reaction times. PMID- 16267594 TI - Bis(amino acid) derivatives of 1,4-diamino-2-butyne that adopt a C2-symmetric turn conformation. AB - 1,4-Diamino-2-butyne was prepared from 1,4-dichloro-2-butyne via 1,4-diazido-2 butyne. Bis(amino acid) derivatives of 1,4-diamino-2-butyne having the general structure (Boc-Xxx-NHCH2C[triple bond])2 (Xxx = Ala, Phe and Met) were prepared and examined by 1H NMR spectroscopy. Using chemical shift, coupling constant and DMSO titration data it is found that these compounds adopt a C2-symmetric turn conformation featuring two intramolecular hydrogen bonds. PMID- 16267595 TI - Lewis-acid assisted cross metathesis of acrylonitrile with functionalized olefins catalyzed by phosphine-free ruthenium carbene complex. AB - The exchange of the PPh3 ligand in the complex [1,3-bis(2,6-dimethylphenyl)4,5 dihydroimidazol-2-ylidene](PPh3)(Cl)2Ru=CHPh (7) for a pyridine ligand at ambient temperature leads to the formation of the stable phosphine-free carbene ruthenium complex [1,3-bis(2,6-dimethylphenyl)4,5-dihydroimidazol-2-ylidene](C5H5N)2(Cl)2 Ru=CHPh (8). The resulted ruthenium complex exhibits highly catalytic activity for the cross metathesis of acrylonitrile with various functionalized olefins under mild conditions, and its activity can be further improved by the addition of a Lewis acid such as Ti(OiPr)4. In the mixture products, the Z-isomer predominates. PMID- 16267596 TI - Synthesis and electronic properties of series of oligothiophene [1,10]phenanthrolines. AB - Organic semiconductors containing metal binding sites within their molecular backbones are of a general interest in organic materials chemistry. In this paper, we describe a straightforward synthetic procedure, which gives access to a series of 2-(oligothienyl)-[1,10]phenanthrolines (nT-phen), 2,9-bis(oligothienyl) [1,10]phenanthrolines (nT-phen-nT) and 2,2'-(oligothienyl)bis [1,10]phenanthrolines (phen-nT-phen). By a Negishi-type cross coupling of 2-iodo [1,10]phenanthroline or 2,9-diiodo-[1,10]phenanthroline with in situ generated alpha-zinc derivatives of different mono-, ter-, and quinquethiophenes we were able to synthesize the corresponding oligothienyl-phenanthrolines in medium to excellent yields. Furthermore, characterization of the optical properties of the new materials indicated that the two subunits, oligothiophene and phenanthroline, are in pi-conjugation. Characterization of the redox properties revealed additional evidence for the role of [1,10]phenanthroline as a pi-bridging unit in the nT-phen-nT series. PMID- 16267597 TI - Exceptionally slow kinetics of the intramolecular quadruplex formed by the Oxytricha telomeric repeat. AB - We examined the stability and kinetics of folding of the Oxytricha telomeric repeat sequence (G4T4)4. Fluorescence melting experiments show that this intramolecular quadruplex, which is more stable in potassium- than sodium containing buffers, shows considerable hysteresis between the melting and annealing profiles, even when heated at a rate of 0.05 degrees C min(-1). Quantitative analysis of this hysteresis, together with temperature-jump relaxation experiments show that the dissociation is exceptionally slow with a half-life of about 10 years at 37 degrees C in the presence of 50 mM K+. The association reaction has a half-life of a few seconds at 37 degrees C, but becomes slower at elevated temperatures consistent with the suggestion that association occurs by a nucleation-zipper mechanism. PMID- 16267598 TI - The incidence of Proteus mirabilis infection increases in patients on treatment but does not trigger disease activity. AB - The aim of this study is to determine whether treatment increases the levels of anti-Proteus antibodies (APA) in patients with rheumatoid arthritis (RA). The blood samples of 32 patients suffering from RA who were recruited in our previous study and continued to participate in our follow-up study were collected after 1 year. Their first and follow-up samples were analysed for the presence of IgG isotype and total immunoglobulins (IgG+IgA+IgM) against Proteus mirabalis (PM) using enzyme-linked immunosorbent assay with two kinds of antigen preparations: whole bacteria and sodium dodecyl sulphate (SDS) lysed bacterial extract. All patients were treated with methotrexate and hydroxychloroquine with adequate dose of non-steroidal anti-inflammatory drug. After 1 year, 11 patients were in clinical remission [erythrocyte sedimentation rate (ESR) less than 30 mm/h and C reactive protein (CRP) less than or equal to 10 mg/l], while the rest of the 21 were in the state of active disease. Correlation and Student's t test were used for statistical analysis. APA titres were significantly elevated in patients after 1 year of therapy. However, the rise was not different between patients who were in clinical remission and those in the state of active disease. APA titre increases in the treatment of RA, and the probable mechanisms are discussed. PMID- 16267599 TI - Cutaneous lesions as presenting symptoms of primary biliary cirrhosis: an undifferentiated connective tissue disease-like onset. AB - We describe two patients with primary biliary cirrhosis (PBC) who presented with specific symptoms mimicking an undifferentiated connective tissue disease (arthromyalgia, fatigue, cutaneous lesions either morbillous-like or urticarial, the latter with an eosinophil infiltrate of upper dermis). Subsequent detection firstly of eosinophilia in the blood and secondarily of antimitochondrial antibodies with results of liver biopsy allowed a diagnosis of asymptomatic PBC. In our cases, a peculiar sign of early stage of PBC was represented also by the eosinophilia in the liver. PMID- 16267600 TI - Adrenal axis functions in patients with familial Mediterranean fever. AB - OBJECTIVE: Familial Mediterranean fever (FMF) is a hereditary disease characterized by recurrent attacks of fever with peritonitis, arthritis, pleuritis or erysipelas-like rash. It is unclear what effects of FMF itself on endocrine system and hormones are. None of the FMF patients without amyloidosis have been reported to have any endocrine disorders, except those who developed colchicine-induced diabetes insipidus. There is a large body of evidence to show that cytokines (IL-1, IL-6 and TNF-alpha) activate the hypothalamic-pituitary adrenal (HPA) axis. We have designated this study to investigate the HPA axis in FMF patients without amyloidosis. METHODS: Twenty-one patients with FMF were included. ACTH stimulation test was performed on the healthy subjects and during attack period in the patients. In the patient group, same test was repeated during remission period. RESULTS: Peak cortisol levels were significantly higher in the attack period than those in the remission period of patients (p<0.05). CONCLUSION: The cytokines play a role on the activation of the HPA axis; we thought the axis would be affected in this disease. The response of cortisol to 250 mug ACTH was significant in attack period when compared with remission period. This result reveals that HPA axis is more activated in an FMF attack. Previous studies suggest that the adrenal hormones increase in acute inflammatory events, and eventually, the changes on these hormones are related to TNF and IL-6 levels. During the FMF attack, HPA axis may be stimulated by cytokines. It seems that HPA axis is regulated normally in FMF patients. PMID- 16267601 TI - Vasculitis of the aortic arch and cardiac valves as the cause of relapsing fever of unknown origin in an elderly, white man. AB - Here, we report the case of fever of unknown origin (FUO) in a 77-year-old white man. The patient presented with a 3-week history of fever (between 38.5 and 39 degrees C) and general malaise. These symptoms had occurred about five to seven times during the past 30 years, and despite repeated hospitalizations, no diagnosis was made. Physical examination did not reveal any specific signs of infection nor did the patient fulfill the criteria for any rheumatic disease including vasculitides. Blood chemistry showed a greatly elevated C-reactive protein (CRP; 158.2 mg/l) and an erythrocyte sedimentation rate >100 mm, indicating an active inflammatory process, and leukocytes were significantly elevated (20,000/mul). Rheumatological parameters showed only nonspecific changes. Finally, a 2-[(18)F]-fluoro-2-deoxy-D: -glucose-positron emission tomography was performed, revealing a markedly enhanced glucose uptake in the ascending aorta and the cardiac valves, indicating vasculitis as the cause of FUO in this patient. Based on this finding, treatment was started with corticosteroids, and 2 days after the initiation of treatment, the patient had normal body temperature, and after 5 days, CRP values had returned to normal. After tapering and final complete removal of steroid treatment, the patient was still free of symptoms, hence no disease-modifying antirheumatic drug therapy was necessary. PMID- 16267602 TI - Long-term mortality outcome in patients with reactive amyloidosis associated with rheumatoid arthritis. AB - It is well established that amyloidosis is a serious clinical complication that can influence the prognosis of patients with rheumatoid arthritis (RA). The purpose of the study was to obtain information on the survival and the hemodialysis (HD) of patients with amyloidosis. Eighty patients (9 men and 71 women) who were diagnosed with amyloidosis by biopsy and definite or classical RA were studied retrospectively. The average duration of RA prior to the diagnosis of amyloidosis was 15.4+/-9.4 years. The average period from the diagnosis of amyloidosis to death was 67.4 months. Forty-nine patients died of the disease (32 cases with HD and 17 cases without HD). Thirty-one patients lived (7 cases with HD and 24 cases without HD). Regarding the survival of these patients, 49 (61.3%) of the 80 patients have died. Survival rate at 28 months was 75%; at 67 months, it was 50%; and at 111 months, it was down to 25%. Mortality rate was 11.9% per year. Survival rate in dialysis at 9.8 months was 75%; at 60.6 months, it dropped to 50%; and at 100.0 months, to 25%. As for patients' survival, high onset age of amyloidosis was the major determining factor for poor survival in these patients (p<0.001). Furthermore, male patients also had poor survival (p=0.07). The long term results were very encouraging to initiate HD in patients with end-stage renal disease due to reactive amyloidosis associated with RA. PMID- 16267603 TI - Comparison of the effects of two antidepressants on exercise performance of the female patients with fibromyalgia. AB - OBJECTIVE: To compare the effects of two antidepressants on exercise performance of the female patients with fibromyalgia. METHODS: Fifteen patients with fibromyalgia participated in this randomized, double-blind crossover study composed of two trials separated by a 2-week washout phase. The pharmacy randomly assigned patients to two groups: (a) placebo (glucose) in the morning and 25 mg amitriptyline at bedtime or (b) 20 mg fluoxetine in the morning and placebo at bedtime. The VO(2max) and Wingate anaerobic test were evaluated. Within 24 h after 15-day medication, the tests were repeated. RESULTS: Peak power and fatigue index increased, and VO(2max) and VT did not change with both fluoxetine and amitriptyline. Mean power significantly increased after fluoxetine but not after amitriptyline. CONCLUSION: Both amitriptyline administration and fluoxetine administration in patients with fibromyalgia have beneficial effects on anaerobic performance. These effects may be important in life quality of the female patients with fibromyalgia. PMID- 16267604 TI - Characterization of relapses in adult idiopathic inflammatory myopathies. AB - The objective of the current report was to determine the relapse rates and characterize the nature of relapses during the disease course of adult patients with idiopathic inflammatory myopathies (IIM). A retrospective cohort study of 53 medical records of patients with polymyositis (PM), dermatomyositis (DM), connective tissue disease (CTD)-associated myositis, and malignancy-associated myositis at an academic rheumatology center was performed. Medical records were reviewed to determine clinical presentation, initial treatment, and clinical follow-up, with an emphasis on relapses. Relapses were defined as a sustained elevation in serum creatine kinase (CK) levels in the absence of an alternative etiology. Patients were followed for an average of 65+/-43 months. All patients received corticosteroids, and 35 patients received additional immunosuppressive medications as part of their initial treatment. Serum CK levels normalized in 51 patients, and muscle strength normalized in 43 patients. Biochemical relapse was observed in 33 patients (65%). Patients with PM and CTD-associated myositis had a higher relapse rate compared to DM and malignancy-associated myositis patients. Multiple relapses were observed in 17 patients. Relapses tended to occur within the first 2 years after treatment initiation and during the tapering phase of treatment. No risk factors were unequivocally identified, although advanced age and increased duration of symptoms prior to treatment initiation had nonsignificant associations with increased risk of relapse. In conclusion, initial treatment of IIM results in a high rate of normalization of serum CK and muscle weakness. However, physicians should be aware of the high rate of relapse in patients with IIM. PMID- 16267605 TI - Management of musculoskeletal complications in endstage renal disease: an update. AB - People treated with prolonged hemodialysis now survive longer and increasingly develop musculoskeletal complications. Significant advances have been made in the management of musculoskeletal manifestations in patients undergoing hemodialysis for endstage renal disease. Noncalcium-based phosphate binders are preferred for the management of hyperphosphatemia. Vitamin D therapy is based on intact parathyroid hormone levels. Newer Vitamin D sterols and calcimimetic agents seem to control secondary hyperparathyroidism without increasing the calcium phosphorous product. Use of biocompatible dialyzers for hemodialysis may delay articular manifestations of beta-2 microglobulin amyloidosis. Beta-2 microglobulin adsorption columns when used along with hemodialysis may lead to lower beta-2 microglobulin levels and improvement in articular manifestations of beta-2 microglobulin amyloidosis. In this review, we focus on the latest advancements in the management of secondary hyperparathyroidism and beta-2 microglobulin amyloidosis. PMID- 16267606 TI - Infliximab in a child with therapy-resistant systemic vasculitis. AB - Treatment of systemic vasculitides is usually based on the use of corticosteroids and other immunosuppressive drugs. We describe a 10-year-old girl with systemic vasculitis resistant to immunosuppressive treatment who had a rapid and impressive response to treatment with infliximab. PMID- 16267607 TI - Coexistence of anomalous origin of the coronary arteries and severe aortic regurgitation in Marfan syndrome. AB - A case report of abnormal origin of the left and right coronary arteries from the right sinus of valsalva with severe aortic regurgitation in Marfan syndrome is presented in this paper. PMID- 16267608 TI - Cytomegalovirus-induced interstitial pneumonitis in a patient with dermatomyositis. AB - Dermatomyositis (DM) patients might present with pulmonary involvement as the first manifestation or during the follow-up period. It is sometimes difficult to determine whether the clinical manifestations related with pulmonary involvement are due to DM or to an infectious process. We report a case of DM patient who developed interstitial pneumonitis induced by cytomegalovirus (CMV) while receiving immunosuppressive treatment. To the best of our knowledge, this is the second report in the literature of interstitial pneumonitis secondary to CMV infection in a patient with DM. PMID- 16267609 TI - Quality of life and life satisfaction in patients with Behcet's disease: relationship with disease activity. AB - Quality of life (QoL) and life satisfaction (LS) are important outcome factors in chronic inflammatory conditions such as Behcet's disease (BD). The aim of this study was to investigate QoL and LS in patients with BD and determine the relationship with disease activity. Forty-one patients with BD and 40 control subjects were involved in the study. Demographic properties were obtained. Disease activity was assessed by Turkish version of BD Current Activity Form (BDCAF) in BD patients. QoL and psychological well-being were assessed by Nottingham Health Profile (NHP) and Life Satisfaction Index (LSI), respectively, in both patients and control groups. The related disease activity measures of QoL and LS were determined. Twenty-two male and 19 female BD patients with a mean age of 33.3+/-9.3 years and 20 male and 20 female control subjects with a mean age of 33.3+/-4.1 years were involved. According to BDCAF, no patient had central nervous system involvement. Thirty-four patients had headache, 33 patients had fatigue, 30 patients had articular involvement, 29 had mucocutaneous lesions, 27 had gastrointestinal involvement, 21 patients had ocular involvement, and 7 patients had vascular involvement. The scores of all dimensions of NHP were significantly higher and the mean score of LSI was significantly lower in BD patients than in control subjects (p<0.001). Correlation analysis indicated that the scores of fatigue, joint involvement, and oral ulcers were the most related factors for physical domains of NHP, whereas joint involvement and genital ulcers were the most related activity measures for psychosocial subscales of NHP. LS was found to be most related with the scores of patient's and physician's impression of disease activity and joint involvement. In conclusion, patients with BD have impaired QoL and disturbed psychological well-being. Current management strategies focusing on fatigue, arthralgia, mucocutaneous lesions, and efforts to measure psychosocial aspects and symptoms of the patients by their point of view will help to improve QoL and raise the LS in patients suffering from BD. PMID- 16267610 TI - Ochronotic spondyloarthropathy: spinal involvement resembling ankylosing spondylitis. AB - Ochronotic spondyloarthropathy is a rare metabolic disease with the musculoskeletal manifestations of alkaptonuria. Ochronotic arthropathy patients may have spinal abnormalities similar to ankylosing spondylitis (AS). The proof of sacroiliac involvement or bamboo spine appearance is not sufficient either for diagnosis of ankylosing spondilitis or exclusion of ochronosis. In this report, the case of a 54-year-old woman having ochronosis, with clinically more recognizable axial arthropathy resembling AS, is presented, and the history, clinical presentation, diagnostic techniques, and distinctive diagnosis are reviewed. PMID- 16267611 TI - Morphologic and immunophenotypic markers as surrogate endpoints of tamoxifen effect for prevention of breast cancer. AB - Prevention trials using incidence or mortality as endpoints require a large number of participants and long follow-up. Trials using biomarkers as endpoints would potentially require fewer participants, less time, and significantly less resources to test promising new agents for breast cancer prevention. To test this idea, a randomized trial of tamoxifen for 1 year versus observation for 1 year was conducted to determine whether tamoxifen can cause regression of hyperplastic breast tissue, whether it changes the biomarker phenotype of premalignant disease or normal breast epithelium, and if biomarkers can be used as early surrogate indicators of response to tamoxifen. Women were identified by having an abnormal mammogram and ductal hyperplasia diagnosed by core needle biopsy. Image-directed needle biopsy was repeated in the same site of the breast after 1 year. Approximately 3000 women were screened, and 265 were eligible. Sixty-three women were randomized and paired biopsies from 45 subjects were available for analysis. There was no evidence of substantial regression of hyperplasia--fewer samples showed hyperplasia at 1 year follow-up, but this was seen in both untreated and tamoxifen-treated women. There were trends for reductions in ER and PgR and trends for increases in bcl-2 in normal and hyperplastic tissue in the tamoxifen treated arm, though these changes did not reach statistical significance. Proliferation, determined by Ki67 staining, was not significantly changed. Clinical trials of this type are difficult to carry out and modifications in trial design are needed to make this process more efficient. PMID- 16267612 TI - Prevalence of the NOD2 3020insC mutation in aggregations of breast and lung cancer. AB - Both breast and lung cancers are common malignancies and within the context of known genetic predispositions to breast cancer, no association has been made in linking the two diseases together. This does not exclude the possibility that such associations may exist that lie outside the known high-risk breast cancer families. To examine the likelihood of common genetic factors that could influence the risk of disease, two sets of consecutively collected tumor groups were examined for the 3020insC mutation in the NOD2/CARD15 gene. A total of 4107 consecutively collected breast cancer patients were assessed for the prevalence of the 3020insC mutation and compared to a consecutively collected series of 389 lung cancer patients and 2068 control samples. The results revealed that a proportion of breast cancer patients who had a first or a second degree relative diagnosed with lung cancer were more likely to harbour a change in NOD2/CARD15 compared to patients who had no relatives affected by lung cancer. Furthermore, this difference appeared to be specific to the breast and lung cancer subgroup since there was no difference in the frequency of the 3020insC allele in the consecutively collected lung cancer patients. In conclusion, it appears that the 3020insC mutation of the NOD2/CARD15 gene may be a genetic predisposing factor for aggregations of breast and lung cancer. PMID- 16267613 TI - Polymorphisms of the promoter regions of matrix metalloproteinases genes MMP-1 and MMP-9 in breast cancer. AB - PURPOSE: Matrix metalloproteinases play a crucial role in the cancer invasion and metastasis, angiogenesis and tumorigenicity. A single guanine insertion--the 1G/2G polymorphism in the promoter of the matrix metalloproteinase 1 (MMP-1) gene creates a binding site for the transcription factor AP-1 and thus may affect the transcription level of MMP-1. The C-->T substitution at the polymorphic site of the MMP-9 gene promoter results in a higher transcription activity of the T allelic promoter trough the loss of binding site for a repressor protein. The aim of this work was to investigate the influence of 1G/2G and C-->T polymorphisms on the MMP-1 and MMP-9 level and therefore on the occurrence and progression of breast cancer. EXPERIMENTAL DESIGN: We investigated the distribution of genotypes and frequency of alleles of the 1G/2G and C-->T polymorphisms for 270 patients with breast cancer and 300 healthy women served as control. The genotypes were determined by RFLP-PCR. Additionally, we estimated the level of MMP-1 and MMP-9 antigens in tumor samples and normal breast tissue using ELISA. RESULTS: The levels of MMP-1 in tumor samples of node positive patients ware significantly higher than in samples of node negative patients (p<0.05). Increased level of MMP 9 correlates with Bloom-Richardson grading III (p<0.05), increased tumor size (p<0.05) and absence of estrogen and progesterone receptors (p<0.01). Additionally, both MMP-1 and MMP-9 levels were higher in tumor than in the normal breast tissue. We showed the higher risk of metastasis development in lymph node for the 2G/2G genotype (OR=2.14; CI 95% 1.24;3.69) and the 2G allele carriers (OR=1.68; CI 95% 1.19;2.39). We found correlation between the T allele (OR=2.61; CI 95% 1.33;4.87), 2G (OR=2.58; CI 95% 1.35;4.91) and malignance. CONCLUSION: The results suggest that MMP-1 is responsible for the local invasion and MMP-9 is associated with the malignance and the growth of the tumor. We suggest that the 2G allele of the 1G/2G MMP-1 gene polymorphism may be associated with the lymph node metastasis in patients with breast cancer and therefore it can be considered as a progression marker in this disease. PMID- 16267614 TI - Cytosolic levels of TFF1/pS2 in breast cancer: Their relationship with clinical pathological parameters and their prognostic significance. AB - BACKGROUND: The Trefoil Factor 1 (TFF1/pS2), a peptide consisting of 60 amino acids, is the most abundant estrogen-induced messenger RNA present in MCF-7 breast cancer cells. The objective of this work was to evaluate the cytosolic TFF1 content in breast carcinomas, its possible relationship with different clinical-pathological parameters, and its potential prognostic significance and predictive value. METHODS: Cytosolic TFF1 levels were examined by immunoradiometric assay in 1031 patients with invasive breast cancer. The median follow-up period was of 50 months. RESULTS: There was a wide variability of cytosolic TFF1 levels in tumors (0.9-743.2 ng/mg protein). Statistical analysis showed that TFF1 levels were significantly higher in premenopausal patients (p = 0.001), as well as in tumors showing any of the following characteristics: good differentiation (p = 0.0001), ER and PgR positivity (p = 0.0001 and p = 0.001, respectively), diploidy (p = 0.045) and a high S-phase fraction (p = 0.001). In addition, the presence of high intratumoral TFF1 levels (cut-off: 2 ng/mg protein) was independently associated with a shorter overall survival in the group of patients as a whole (p = 0.001) as well as in the subgroup with node negative breast cancer (p = 0.0004). Likewise, high intratumoral TFF1 levels were associated with a more prolonged overall survival in patients who received adjuvant tamoxifen (p = 0.004). CONCLUSIONS: In breast cancer patients, intratumoral TFF1 levels are associated with a better clinical outcome, especially in those with node-negative tumors. In addition, TFF1 levels have a low but significant predictive value in regards to response to adjuvant therapy with tamoxifen. PMID- 16267615 TI - Multicenter phase II study of trastuzumab in combination with epirubicin and docetaxel as first-line treatment for HER2-overexpressing metastatic breast cancer. AB - The primary objective of study is to evaluate cardiac safety of trastuzumab in combination with epirubicin and docetaxel. HER2-overexpressing metastatic breast cancer patients were enrolled in a two-stage, multicenter phase II trial with weekly trastuzumab (4 and then 2 mg/kg) with epirubicin and docetaxel (either 75 mg/m(2)) on day 1 every 3 weeks. After eight courses of chemotherapy, trastuzumab was continued as a single agent. To assess cardiotoxicity, patients were evaluated for left ventricular ejection fraction (LVEF) at baseline, every two cycles during chemotherapy and trastuzumab, and every 3 months during trastuzumab alone. Cardiotoxicity was defined as signs and/or symptoms of congestive heart failure (CHF) and/or an absolute decrease in LVEF of >or=20 units or a decline to or = 1.15 of bilateral SN echogenic sizes, estimated by division of larger size by smaller size, was found in 69% of PDD patients but only 20% of DLB patients. Combination of SN echogenic sizes, asymmetry indices and onset age discriminated PDD from DLB with a sensitivity of 96%, a specificity of 80% and a positive predictive value of 93%. TCS of brainstem raphe, thalami, lenticular nuclei, caudate nuclei and ventricle widths did not discriminate between DLB and PDD. Compared with PDnD patients, DLB and PDD patients exhibited significantly larger widths of third ventricle and of frontal horns. In PDD patients, scores on the Unified Parkinson's Disease Rating Scale correlated with widths of third ventricle and of frontal horns. CONCLUSIONS: SN hyperechogenicity is typical for PDD and DLB. However, size, asymmetry and relation of SN hyperechogenicity to age at disease onset discriminate PDD from DLB. PMID- 16267639 TI - Hyperhomocysteinemia is associated with volumetric white matter change in patients with small vessel disease. AB - BACKGROUND: Hyperhomocysteinemia is associated with cerebral small vessel disease (SVD). We examined the relationship between homocysteine and 1) volumetric measure of white matter change (WMC), 2) silent brain infarcts, 3) cerebral atrophy on MRI and 4) cognition on a consecutive cohort of patients with stroke associated with SVD. SUBJECTS AND METHODS: Fifty-seven patients consecutively admitted to the Acute Stroke Unit in a university hospital due to stroke associated with SVD were recruited and assessed three months after the stroke. Non-fasting homocysteine was obtained. Using MRI, the number of infarcts, volume of WMC and cerebral atrophy were measured. General cognitive functions were assessed using the Mini Mental State Examination and Alzheimer's disease Assessment Scale. Mattis Dementia Rating Scale - Initiation/Perseveration subset was used to assess executive cognitive functions. RESULTS: Hyperhomocysteinemia (> or = 14.88 micromol/L) significantly accounted for the volume of WMC on MRI in a multivariate stepwise regression model (adjusted R(2)=0.058, p <0.05) after adjustment for age and folate level. Patients in the highest quartile of WMC volume had significantly higher levels of homocysteine than those in lowest quartile (p <0.001). No significant relationship was found between homocysteine and silent brain infarcts, cerebral atrophy and performance on psychometric tests. CONCLUSION: Hyperhomocysteinemia is associated with volumetric measure of WMC among patients with SVD. The role of homocysteine in the development of silent brain infarcts and cerebral atrophy as previously reported cannot be ascertained in this study. No direct relationship was found between homocysteine and cognitive functions. PMID- 16267641 TI - Verbal and spatial working memory in autism. AB - Verbal and spatial working memory were examined in high-functioning children, adolescents, and adults with autism compared to age and cognitive-matched controls. No deficit was found in verbal working memory in the individuals with autism using an N-back letter task and standardized measures. The distinction between the N-back task and others used previously to infer a working memory deficit in autism is that this task does not involve a complex cognitive demand. Deficits were found in spatial working memory. Understanding the basis for the dissociation between intact verbal working memory and impaired spatial working memory and the breakdown that occurs in verbal working memory as information processing demands are increased will likely provide valuable insights into the neural basis of autism. PMID- 16267642 TI - Gastrointestinal factors in autistic disorder: a critical review. AB - Interest in the gastrointestinal (GI) factors of autistic disorder (autism) has developed from descriptions of symptoms such as constipation and diarrhea in autistic children and advanced towards more detailed studies of GI histopathology and treatment modalities. This review attempts to critically and comprehensively analyze the literature as it applies to all aspects of GI factors in autism, including discussion of symptoms, pathology, nutrition, and treatment. While much literature is available on this topic, a dearth of rigorous study was found to validate GI factors specific to children with autism. PMID- 16267640 TI - Clinical phenotypic heterogeneity of Alzheimer's disease associated with mutations of the presenilin-1 gene. AB - It is now 10 years since the first report of mutations in the presenilin genes that were deterministic for familial autosomal dominant Alzheimer's disease. The most common of these mutations occurs in the presenilin-1 gene (PSEN1) located on chromosome 14. In the ensuing decade, more than 100 PSEN1 mutations have been described. The emphasis of these reports has largely been on the novelty of the mutations and their potential pathogenic consequences rather than detailed clinical, neuropsychological, neuroimaging and neuropathological accounts of patients with the mutation. This article reviews the clinical phenotypes of reported PSEN1 mutations, emphasizing their heterogeneity, and suggesting that other factors, both genetic and epigenetic,must contribute to disease phenotype. PMID- 16267645 TI - The determination of total Se in urine and serum by graphite furnace atomic absorption spectrometry using Ir as permanent modifier and in situ oxidation for complete trimethylselenonium recovery. AB - The present work evaluated the use of iridium (Ir) as permanent modifier for the determination of total selenium in urine and serum by graphite furnace atomic absorption spectrometry. Concerning urine, the presence of trimethylselenonium (TMSe(+)) was especially considered. Pyrolysis and atomization temperatures of 1,000 and 2,100 degrees C, respectively, were used. For nondigested urine and serum samples, 0.2% v/v HNO(3) and Triton X-100 were used as diluents, respectively, and the same initial platform Ir treatment was effective for up to 1,100 atomization cycles. Good precision [less than 5% relative standard deviation (RSD)] can be achieved with the proposed method. Low TMSe(+) recovery was observed for nondigested urine samples. Thus, if this species is to be considered in urine analysis, a previous external mineralization step was found to be necessary. Alternatively, an in situ oxidation treatment was developed. Detection limits of 8, 10, and 7 mug l(-1) were obtained after dilution, microwave-assisted digestion, and in situ oxidation procedures, respectively. The accuracy of the method was validated by the analysis of certified reference or commercial quality control materials and spiked samples. PMID- 16267646 TI - Determination of multiclass pesticides in food commodities by pressurized liquid extraction using GC-MS/MS and LC-MS/MS. AB - Pressurized liquid extraction (PLE) was applied to the simultaneous extraction of a wide range of pesticides from food commodities. Extractions were performed by mixing 4 g of sample with 4 g of Hydromatrix and (after optimization) a mixture of ethyl acetate:acetone (3:1, v/v) as extraction solvent, a temperature of 100 degrees C, a pressure of 1000 psi and a static extraction time of 5 min. After extraction, the more polar compounds were analyzed by liquid chromatography (LC), and the apolar and semipolar pesticides by gas chromatography (GC); in both cases LC and GC were coupled with mass spectrometry in tandem (MS/MS) mode. The overall method (including the PLE step) was validated in GC and LC according to the criteria of the SANCO Document of the European Commission. The average extraction recoveries (at two concentration levels) for most of the analytes were in the range 70-80%, with precision values usually lower than 15%. Limits of quantification (LOQ) were low enough to determine the pesticide residues at concentrations below or equal to the maximum residue levels (MRL) specified by legislation. In order to assess its applicability to the analysis of real samples, aliquots of 15 vegetable samples were processed using a conventional extraction method with dichloromethane, and the results obtained were compared with the proposed PLE method; differences lower than 0.01 mg kg(-1) were found. PMID- 16267647 TI - Development of a new biosensor for superoxide radicals. AB - A superoxide dismutase (SOD) biosensor for determination of superoxide radicals has been developed by immobilization of superoxide dismutase within gelatin (G) on a Pt electrode surface. The properties of the biosensor have been investigated and optimum conditions-enzyme concentration, glutaraldehyde concentration, and pH were determined. The response of the G-SOD biosensor was proportional to O2*- concentration and the detection limit was 0.01 mmol L(-1) at a signal-to-noise ratio of 3. The biosensor retained 89% and 60% of its sensitivity after use for three and four weeks, respectively. Immobilization of SOD on gelatin provides a biocompatible microenvironment around the enzyme and stabilizes the activity of the enzyme very efficiently. The superoxide dismutase biosensor was used to determine the antioxidant properties of acetylsalicylic acid-based drugs and the anti-radical activity of healthy and cancerous human brain tissues. PMID- 16267643 TI - Imaging in cell-based therapy for neurodegenerative diseases. AB - Fetal cell transplantation for the treatment of Parkinson's and Huntington's diseases has been developed over the past two decades and is now in early clinical testing phase. Direct assessment of the graft's survival, integration into the host brain and impact on neuronal functions requires advanced in vivo neuroimaging techniques. Owing to its high sensitivity, positron emission tomography is today the most widely used tool to evaluate the viability and function of the transplanted tissue in the brain. Nuclear magnetic resonance techniques are opening new possibilities for imaging neurochemical events in the brain. The ultimate goal will be to use the combination of multiple imaging modalities for complete functional monitoring of the repair processes in the central nervous system. PMID- 16267648 TI - [CT-based assessment score after ventral spondylodesis for thoracolumbar spine fracture]. AB - BACKGROUND: Dorsoventral stabilization is a common procedure to treat thoracolumbar spine fractures. Especially in respect of the evaluation of alternative procedures to autogenous bone graft, a standardized evaluation score for ventral spondylodesis is necessary. PATIENTS AND METHODS: In a group of 44 patients a follow-up CT scan was evaluated with a standardized scoring system by four different independent evaluators (a trauma surgeon, an orthopedic surgeon, and two radiologists). The score is based on the morphologic classification of the region between graft and vertebral body. It allows a classification of the spondylodesis as sufficient, partial, and not sufficient. RESULTS: The statistical evaluation of the classification of the different evaluators shows very good interobserver agreement in monosegmental fusion and good agreement in bisegmental fusion. CONCLUSION: The demonstrated score is easy to handle, does not need special equipment for CT scans, and shows good interobserver agreement in the classification of spinal fusion after ventral spondylodesis for thoracolumbar spine fracture. PMID- 16267650 TI - Treatment option in a SLAP-related ganglion cyst resulting in suprascapular nerve entrapment. AB - INTRODUCTION: Our report shows a rare case of suprascapular nerve palsy due to a SLAP-related ganglion cyst resulting in isolated weakness of the infraspinatus muscle. CASE REPORT: We report on a 31-year old volleyball player with severe shoulder pain. A ganglion cyst was excised in an open procedure and was completely resolved in a postoperative magnetic resonance imaging (MRI). But the patient again had pain and disability 7 months after this procedure. A renewed MRI scan showed a cystic mass in the spinoglenoid notch. An electromyography revealed an isolated lesion of the suprascapular nerve. The patient was treated by shoulder arthroscopy with refixation of a type-II-SLAP-lesion and drainage of the cyst formation. At latest follow-up 29 months after surgery, the patient's pain and shoulder function improved with a constant score of 94 points. A MRI scan documented complete cyst resolution. CONCLUSIONS: Treatment options for ganglion cysts at the spinoglenoid notch are various and can be handled in conservative and operative ways. We believe that the arthroscopic concept with the management of a SLAP lesion as the cause of cyst formation, and the drainage of the ganglion is an effective way with low surgical morbidity that shows good postoperative results. PMID- 16267651 TI - Dynamic function of the fibula. Gait analysis evaluation of three different parts of the shank after fibulectomy: proximal, middle and distal. AB - The purpose of this case study was to investigate the dynamic features of fibular movement to gait pattern by analyzing the gait of individuals with three different parts of the fibula resected. Gait analyses revealed that proximal fibula resection impaired knee stability, whereas distal fibula resection disturbed ankle kinematics significantly. Except a mild secondary quadriceps weakness, middle fibula resection did not cause a significant biomechanical disturbance on gait. PMID- 16267649 TI - C1 inhibitor: molecular and clinical aspects. AB - C1 inhibitor (C1-INH) is a serine protease inhibitor (serpins) that inactivates several different proteases in the complement, contact, coagulation, and fibrinolytic systems. By its C-terminal part (serpin domain), characterized by three beta-sheets and an exposed mobile reactive loop, C1-INH binds, and blocks the activity of its target proteases. The N-terminal end (nonserpin domain) confers to C1-INH the capacity to bind lipopolysaccharides and E-selectin. Owing to this moiety, C1-INH intervenes in regulation of the inflammatory reaction. The heterozygous deficiency of C1-INH results in hereditary angioedema (HAE). The clinical picture of HAE is characterized by bouts of local increase in vascular permeability. Depending on the affected site, patients suffer from disfiguring subcutaneous edema, abdominal pain, vomiting and/or diarrhoea for edema of the gastrointestinal mucosa, dysphagia, and dysphonia up to asphyxia for edema of the pharynx and larynx. Apart from its genetic deficiency, there are several pathological conditions such as ischemia-reperfusion, septic shock, capillary leak syndrome, and pancreatitis, in which C1-INH has been reported to either play a pathogenic role or be a potential therapeutic tool. These potential applications were identified long ago, but controlled studies have not been performed to confirm pilot experiences. Recombinant C1-INH, produced in transgenic animals, has recently been produced for treatment of HAE, and clinical trials are in progress. We can expect that the introduction of this new product, along with the existing plasma derivative, will renew interest in exploiting C1 INH as a therapeutic agent. PMID- 16267653 TI - Differential regulation of vacuolar H+ -ATPase and Na+/H+ exchanger 3 in rat cholangiocytes after bile duct ligation. AB - The cholangiocytes lining the intrahepatic bile ducts modify the primary secretion from the hepatocytes. The cholangiocytes secrete HCO (3)(-) into bile when stimulated with secretin in many species, including man. However, in rats, secretin stimulation neither affects biliary HCO (3)(-) concentration nor bile flow, whereas following bile duct ligation (BDL) it induces hypercholeresis with significant increase of NaHCO(3) concentration. We hypothesized that BDL might affect the expression of cholangiocyte H(+) transporters and thereby choleresis, and determined the expression and localization of the 31 kDa vacuolar type H(+) ATPase (V-ATPase) subunit and of Na(+)/H(+) exchanger NHE3 in the livers of control and BDL rats by real-time PCR, in situ hybridization, immunoblotting, and immunohistochemistry. In controls, secretin had no effect on bile flow, whereas following BDL, secretin increased bile flow approximately threefold. V-ATPase and NHE3 were expressed in control cholangiocytes showing intracellular and apical distribution, respectively. BDL significantly up-regulated V-ATPase mRNA and protein expression and was associated with redistribution to the apical pole in approximately 60% of the cholangiocytes lining the small bile ductules. In contrast, NHE3 expression was significantly down-regulated by BDL at the mRNA and protein level. The data demonstrate expression of V-ATPase in rat cholangiocytes. BDL-induced down-regulation of NHE3 may contribute to a reduction of Na(+) and HCO (3)(-) reabsorption and thus to their net secretion into bile. Apical localization of V-ATPase in cholangiocytes may indicate its involvement in pH regulation and/or HCO (3)(-) salvage to compensate for NHE3 down-regulation in BDL. PMID- 16267654 TI - The cost-effectiveness of dual oral antiplatelet therapy following percutaneous coronary intervention: a Swedish analysis of the CREDO trial. AB - The CREDO trial demonstrated the clinical efficacy of 12-month antiplatelet therapy with clopidogrel compared to standard 28-day treatment with a 27% relative reduction in the combined risk of death, myocardial infarction, or stroke in patients undergoing percutaneous coronary intervention (PCI) and being treated with aspirin. This study evaluated the long-term cost-effectiveness of 12 month vs. 28-day therapy with clopidogrel in Sweden. A Markov model was developed which assumed a hypothetical cohort of patients in a post-PCI state to have certain risks of suffering one of the endpoints of the CREDO trial: stroke, myocardial infarction, or death. The model predicted a mean survival of 12.098 years in the 12-month arm vs. 12.026 in the 28-day arm, an incremental gain of 0.072 life-years. The gain in survival came at a predicted incremental cost of Euro 217, resulting in an incremental cost-effectiveness ratio of Euro 3,022. Thus the predicted cost-effectiveness ratio of long-term treatment with clopidogrel in patients undergoing PCI is well below the threshold values currently considered cost-effective. PMID- 16267652 TI - VASP-dependent regulation of actin cytoskeleton rigidity, cell adhesion, and detachment. AB - Enabled/vasodilator-stimulated phosphoprotein (Ena/VASP) proteins are established regulators of actin-based motility, platelet aggregation, and growth cone guidance. However, the molecular mechanisms involved essentially remain elusive. Here we report on a novel mechanism of VASP action, namely the regulation of tensile strength, contractility, and rigidity of the actin cytoskeleton. Compared to wild-type cells fibroblasts derived from VASP-deficient mice have thicker and more stable actin stress fibres. Furthermore focal adhesions are enlarged, myosin light chain phosphorylation is increased, and the rigidity of the filament supported plasma membrane is elevated about three- to fourfold, as is evident from atomic force microscopy. Moreover, fibronectin-coated beads adhere stronger to the surface of VASP-deficient cells. The resistance of these beads to mechanical displacement by laser tweezers is dramatically increased in an F-actin dependent mode. Cytoskeletal stabilization coincides with slower cell adhesion and detachment, while overall adhesion is increased. Interestingly, many of these effects observed in VASP (-/-) cells are recapitulated in VASP-overexpressing cells, hinting towards a balanced stoichiometry necessary for appropriate VASP function. Taken together, our results suggest that VASP regulates surface protrusion formation and cell adhesion through modulation of the mechanical properties of the actin cytoskeleton. PMID- 16267655 TI - Spanish health benefits for services of curative care. AB - This contribution presents entitlements and benefits, decision criteria, and involved actors for services of curative care in Spain. It describes basic benefits included in the category of curative care defined by the central government and any additional benefits that some autonomous communities (ACs) have included to enlarge their own basket. It is concluded that there is no specific and explicit benefit catalogue. As no user charges exist for this category, waiting times serve as the main cost containment tool. There is a need for further legislation, as inequalities may increase across the territory as a matter of fact. Inequalities in access to health care resources between ACs are not due to differences in health baskets but mainly to the availability of technologies. PMID- 16267656 TI - Benefits and entitlements in the Hungarian health care system. AB - This contribution considers entitlements and benefits in the Hungarian health care system. After a brief introduction to the organizational structure of the system the decision-making processes are discussed in detail, including the most important actors, types and pieces of legislation, formal structures, decision making criteria, and outputs in terms of benefit catalogues. Within the two main public financing systems (social insurance and tax-funded services) there are four types of regulatory regimes: (a) traditional political decision making, (b) price negotiations, (c) updating of classification systems for payment purposes, and (d) the procedure for the inclusion of registered medicines in the scope of the social health insurance system. As an example we discuss the benefit regulations and benefit catalogues in the category of services of curative care (HC.1) of the OECD classification of health services. PMID- 16267657 TI - The "Health Benefit Basket" in France. AB - The French "Health Benefit Basket" is defined principally by positive lists of reimbursed goods and services; however, global budget-financed hospital-delivered services are more implicitly defined. The range of reimbursable curative care services is defined by two coexisting positive lists/fee schedules: the Classification Commune des Actes Medicaux (CCAM) and the Nomenclature Generale des Actes Professionnels (NGAP). The National Union of Health Insurance Funds has been updating these positive lists since August 2004, with the main criterion for inclusion being the proposed procedure's effectiveness. This is assessed by the newly created High Health authority (replacing the former ANAES). In addition, complementary health insurers are consulted in the inclusion process due to their important role in French health care financing. PMID- 16267658 TI - Hospital development plans: a new tool to break ground for strategic thinking in Tanzanian hospitals. AB - Tanzanian hospitals suffer from underfunding and poor management. In particular, planning and strategic thinking need improvement. Cultural values such as subordination, risk aversion, and high time preference, together with a long history of socialist government, result in lack of responsibility, accountability, and planning. This has been addressed by the health sector reform with its focus on decentralization, strengthened by the introduction of basket funding facilitated by the Comprehensive Council Health Plans. As a consequence of this the next logical step is to improve the authority of regional and district hospitals in the use of their resources by introducing hospital development plans. These strategic plans were introduced as tools of strategic planning in 2001 by the Kreditanstalt fur Wiederaufbau in close collaboration with the Tanzanian Ministry of Health, binding the release of rehabilitation funds to presentation of a strategic hospital plan. This study examines the rationale and content of hospital development plans. Initial experiences are discussed. The quality of presented plans has steadily improved, but there is a tendency for hospitals with a close connection to development partners to present well prepared reports while other hospitals have severe problems fulfilling the requirements. For many hospitals it is in fact the first time that they have had to define their functions and future role, thus breaking ground for strategic thinking. PMID- 16267659 TI - Cost-effectiveness of outpatient treatment in depressive patients with escitalopram in Germany. AB - We investigated the cost-effectiveness of escitalopram (10 mg daily dose) vs. venlafaxine XR (75 mg daily dose) in a German outpatient setting for the treatment of unipolar depression (MADRS score 20-34) over a period of 70 days. To assess the cost effectiveness of the two substances we combined data from physician's surveys and clinical response data; cost-effectiveness was calculated using a Markov model. In a second step we considered the therapeutic decisions of the attending physicians. Cost-effectiveness was indicated as costs per successfully treated patient. Escitalopram demonstrated a more favorable cost effectiveness ratio than venlafaxine XR. The analysis of treatment patterns showed that attending physicians intervene fairly early if the chosen therapy is ineffective. Additional costs for the use of venlafaxine XR over those of escitalopram were estimated from Euro 7,446 to Euro 9.836 per successfully treated per patient. Hence escitalopram may be a cost-effective alternative to venlafaxine XR in outpatient care setting in Germany. PMID- 16267660 TI - Kinetics and mechanistic studies of the reactions of metleghemoglobin, ferrylleghemoglobin, and nitrosylleghemoglobin with reactive nitrogen species. AB - It is now established that nitrogen monoxide is produced not only in animals but also in plants. However, much less is known about the pathways of generation and the functions of NO. in planta. One of the possible targets of NO. is leghemoglobin (Lb), the hemoprotein found in high concentrations in the root nodules of legumes that establish a symbiosis with nitrogen-fixing bacteria. In analogy to hemoglobin and myoglobin, we have shown that different forms of Lb react not only with NO. but also with so-called reactive nitrogen species derived from it, among others peroxynitrite and nitrite. Because of the wider active-site pocket, the rate constants measured in this work for NO. and for nitrite binding to metLb are 1 order of magnitude larger than the corresponding values for binding of these species to metmyoglobin and methemoglobin. Moreover, we showed that reactive nitrogen species are able to react with two forms of Lb that are produced in vivo but that cannot bind oxygen: ferrylLb is reduced by NO. and nitrite, and nitrosylLb is oxidized by peroxynitrite. The second-order rate constants of these reactions are on the order of 10(2), 10(6), and 10(5) M-1 s-1, respectively. In all cases, the final reaction product is metLb, a further Lb form that has been detected in vivo. Since a specific reductase is active in nodules, which reduces metLb, reactive nitrogen species could contribute to the recycling of these inactive forms to regenerate deoxyLb, the oxygen-binding form of Lb. PMID- 16267661 TI - Oxyleghemoglobin scavenges nitrogen monoxide and peroxynitrite: a possible role in functioning nodules? AB - It has been demonstrated that the NO* produced by nitric oxide synthase or by the reduction of nitrite by nitrate reductase plays an important role in plants' defense against microbial pathogens. The detection of nitrosyl Lb in nodules strongly suggests that NO* is also formed in functional nodules. Moreover, NO* may react with superoxide (which has been shown to be produced in nodules by various processes), leading to the formation of peroxynitrite. We have determined the second-order rate constants of the reactions of soybean oxyleghemoglobin with nitrogen monoxide and peroxynitrite. At pH 7.3 and 20 degrees C, the values are on the order of 10(8) and 10(4) M-1 s-1, respectively. In the presence of physiological amounts of CO2 (1.2 mM), the second-order rate constant of the reaction of oxyleghemoglobin peroxynitrite is even larger (10(5) M-1 s-1). The results presented here clearly show that oxyleghemoglobin is able to scavenge any NO* and peroxynitrite formed in functional nodules. This may help to stop NO* triggering a plant defense reaction. PMID- 16267662 TI - Synthesis, structure and biological activity of a new and efficient Cd(II)-uracil derivative complex system for cleavage of DNA. AB - The new complex formed by Cd(II) and the 1:2 Schiff-base-type ligand 2,6-bis[1-(4 amino-1,2,3,6-tetrahydro-1,3-dimethyl-2,6-dioxopyrimidin-5-yl)imino]ethylpyridine (DAPDAAU) has been chemically and structurally characterized by X-ray diffraction: the ion Cd(II) is surrounded by six nitrogen atoms from two DAPDAAU ligands which coordinates each one in a tridentate fashion through the pyridine ring (N1) and both azomethine nitrogen atoms (N5). The interaction of the Cd(II) complex (compound I) with calf-thymus DNA as observed by circular dichroism spectroscopy suggests the initial unwinding of the DNA double helix strongly depends on increasing incubation times and metal-to-nucleic acid molar ratios. Electrophoretic experiments indicate that the cadmium complex induces cleavage of the plasmid pBR322 DNA to give ulterior nicking and shortening of this molecule, as a result of the complex binding to DNA, resulting in the conclusion that compound I behaves as a chemical nuclease. Cytotoxic activity of the Cd(II) complex against selected different human cancer cell lines is specific and increases with increasing concentration of the metal compound; this fact indicates the potential antitumor character of the complex. When the culture medium is supplemented with compound I, a remarkable inhibition of the growing cell is observed, important cell degeneration appears before 48 h and abundant precipitates are formed that correspond to cell residues and denatured proteins. PMID- 16267664 TI - Observation of contrast enhancement in the cochlear fluid space of healthy subjects using a 3D-FLAIR sequence at 3 Tesla. AB - In animals, the enhancement of perilymph in the cochlea has been reported using 1.25 mmol/kg of Gd-DTPA, allowing the separate visualisation of perilymph and endolymph for the diagnosis of Meniere's disease. The purpose of this study was three-fold: (1) to determine the optimal timing for detecting cochlear fluid enhancement using 3D-FLAIR (fluid-attenuated inversion recovery) after intravenous administration of 0.1 mmol/kg of Gd-DTPA in healthy human subjects; (2) to examine the reliability of enhancement in multiple healthy subjects; and (3) to investigate whether endolymph and perilymph space can be visually discriminated. In two healthy subjects, 3D-FLAIR images were obtained before, immediately after and 2 h, 4 h and 6 h after the injection. Three more healthy subjects were scanned before and 4 h after the injection. In all four ears of the initial two subjects, cochlear fluid was found to be most intensely enhanced 4 h after the injection. In all of the additional three subjects, the cochlear fluid signal had increased after 4 h from injection. However, visual differentiation of endolymph and perilymph fluid could not be achieved. Using 3D-FLAIR and Gd-DTPA, cochlear fluid enhancement can be observed in healthy human ears, even with a single dose of contrast-medium injection. PMID- 16267663 TI - Ab initio model studies of copper binding to peptides containing a His-His sequence: relevance to the beta-amyloid peptide of Alzheimer's disease. AB - Two of the defining hallmarks of Alzheimer's disease (AD) are deposits of the beta-amyloid peptide, Abeta, and the generation of reactive oxygen species, both of which may be due to the Abeta peptide coordinating metal ions. The Cu2+ concentrations in cores of senile plaques are significantly elevated in AD patients. Experimental results indicate that Abeta1-42 in particular has a very high affinity for Cu2+, and that His13 and His14 are the two most firmly established ligands in the coordination sphere of the copper ion. Quantum chemical calculations using the unrestricted B3LYP hybrid density functional method with the 6-31G(d) basis set were performed for geometries, zero point energies and thermochemistry. The effects of solvation were accommodated using the CPCM method. The enthalpies were calculated with the 6-311+G(2df,2p) basis set. Calculations show that when Cu(H2O)(4)2+ combines with the model compound 1 (3-(1H-imidazol-5-yl)-N-[2-(1H-imidazol-5-yl)ethyl] propanamide) in the aqueous phase, the most stable binding site involves the Npi atoms of His13 and His14 as well as the carbonyl of the intervening backbone amide group. These structures are fairly rigid and the implications for conformational changes to the Abeta backbone are discussed. In solution at pH=7, Cu2+ promotes the deprotonation and involvement in the binding of the backbone amide nitrogen in a beta-sheet like structure. This geometry does not induce strain in the peptide backbone, making it the most likely representation of that portion of the Cu2+-Abeta complex monomer in aqueous solution. PMID- 16267665 TI - Clinical evaluation of bone-subtraction CT angiography (BSCTA) in head and neck imaging. AB - Fifty-one patients were examined with bone subtraction CT angiography (BSCTA). Data were acquired on 4-and 64-slice spiral CT systems. The post-processing method is based on fully automatic registration of non-enhanced and contrast enhanced CT data and subsequent selective bone removal. Vascular structures and brain tissue are retained with the original CTA noise level. Image quality and delineation of the pathologic process were assessed and artifacts introduced by the bone removal process recorded. The bone subtraction algorithm worked successfully in all examinations. The processing time was 6 min on average. Image quality was rated excellent in 20 (39%), good in 26 (51%) and acceptable in 5 (10%) patients. Ophthalmic arteries were visible in 12 (24%) patients bilaterally, in 13 (25%) patients unilaterally and in 26 (51%) patients at least at the origin. BSCTA improved visualization of the infraclinoid ICA and the vertebral arteries. The depiction of stenosis of the extracranial ICA and supraclinoid aneurysms was not significantly improved. In patients with suspicion of sinus thrombosis, BSCTA and conventional CTA yielded similar results. To conclude, BSCTA improves the visualization of vessels with close contact to bone and can improve the diagnostic accuracy and therapy planning of infraclinoid aneurysms. PMID- 16267666 TI - Radiofrequency ablation of chondroblastoma using a multi-tined expandable electrode system: initial results. AB - The standard treatment for chondroblastoma is surgery, which can be difficult and disabling due to its apo- or epiphyseal location. Radiofrequency (RF) ablation potentially offers a minimally invasive alternative. The often large size of chondroblastomas can make treatment with plain electrode systems difficult or impossible. This article describes the preliminary experience of RF treatment of chondroblastomas with a multi-tined expandable RF electrode system. Four cases of CT guided RF treatment are described. The tumour was successfully treated in all cases. In two cases, complications occurred; infraction of a subarticular chondroblastoma in one case and cartilage and bone damage in the unaffected compartment of a knee joint in the other. Radiofrequency treatment near a joint surface threatens the integrity of cartilage and therefore long-term joint function. In weight-bearing areas, the lack of bone replacement in successfully treated lesions contributes to the risk of mechanical failure. Multi-tined expandable electrode systems allow the treatment of large chondroblastomas. In weight-bearing joints and lesions near to the articular cartilage, there is a risk of cartilage damage and mechanical weakening of the bone. In lesions without these caveats, RF ablation appears promising. The potential risks and benefits need to be evaluated for each case individually. PMID- 16267667 TI - Increasing anastomosis safety and preventing abdominal adhesion formation by the use of polypeptides in the rat. AB - BACKGROUND AND AIMS: Postoperative adhesions can potentially be reduced using different anti-adhesive agents, though these drugs tend to compromise healing of an intestinal anastomosis. No method that significantly increases anastomosis safety is known at present. The aim of the study was to develop a concept of preventing postoperative adhesions using differently charged bioactive polypeptides, also considering healing and safety of an intestinal anastomosis. METHODS: An ileocolic anastomosis was performed under both "clean" and "septic" conditions in the rat. The treatment group received intraperitoneal poly-L-lysine and poly-L-glutamate, while controls received sodium chloride. Abdominal adhesions, anastomosis leakage and burst pressure were analysed after 1, 3, 5 and 7 days in the clean anastomosis model and after 7 days in the septic model. RESULTS: A significant decrease (p<0.01) in the amount of adhesions was seen in animals treated with polypeptides after 1, 3 and 5 days, while no difference was seen after 7 days. The anastomosis demonstrated a significantly higher burst pressure as evaluated at days 1 and 3 (p<0.05 and p<0.01, respectively) in the polypeptide-treated animals, while no difference was seen between the groups at day 5 or 7. CONCLUSION: The use of differently charged polypeptides administered intraperitoneally after surgery resulted in a significant decrease in the extent of postoperative adhesions. Furthermore, an increase in intestinal anastomosis safety, based on improved burst pressure during the first 3 days, i.e. the critical period during the healing process, was noted. No adverse effects were seen in surgery during septic conditions. PMID- 16267668 TI - Isolation and cultivation of neuronal precursor cells from the developing human enteric nervous system as a tool for cell therapy in dysganglionosis. AB - BACKGROUND: The human enteric nervous system (ENS) descends from migrating neural crest cells (NCC) and is structured into different plexuses embedded in the gastrointestinal tract wall. The development of this entity strongly depends on the supply of an appropriate support with trophic factors during organogenesis. The lack of important factors, such as glial cell line-derived neurotrophic factor, leads to severe disturbances in the ENS and, thus, to motility disorders in children. The isolation of neuronal precursor cells as well as their transplantation after expansion in vitro is therefore a hopeful new approach concerning all forms of dysganglionosis in children. METHODS: We therefore established a way to isolate and expand precursor cells from the developing and postnatal human ENS. Bowel samples were obtained from human fetuses and children (from the 9th week of gestation to 5 years postnatal). Myenteric plexus was isolated by enzymatical digestion and cultivated until spheroid aggregates, the so-called neurospheres, developed. These neurospheres could be differentiated and also be transplanted after dissociation into aganglionic bowel in vitro. RESULTS: Enteric neurospheres could be grown from different gestational ages, including postmortem material. Undifferentiated proliferating precursor cells were kept in culture for up to 72 days and could be differentiated in neurons and glial cells in vitro. CONCLUSION: The first results using isolated enteric neurospheres in aganglionic bowel are quite promising and are a basis to develop an appropriate cell therapy for all kinds of dysganglionosis, especially for cases where a surgical approach is not sufficient or not even possible. PMID- 16267669 TI - The duration of postoperative ileus after elective colectomy is correlated to surgical specialization. AB - AIM: Postoperative ileus is an important factor of complications following gastrointestinal procedures. Its pathophysiology and the parameters, which may impact on its duration, remain unclear. The aim of this study was to measure the role of various clinical determinants on restoration of intestinal function after elective colorectal surgery. METHODS: From July 2002 to September 2003, all patients who underwent laparotomy for colectomy (laparoscopic resections excluded) with either an ileotransverse, colocolic, or high colorectal anastomosis were entered in this prospective study. The intervals in hours between the end of the surgical procedure and passing of flatus (PG) and passing of stool (PS) were recorded by an independent investigator. PG and PS were eventually correlated with the following parameters: type of colectomy, early removal of nasogastric tube (NGT), mechanical bowel preparation (MBP), type of underlying disease, systemic administration of opiates, and surgical training (colorectal fellowship or other). RESULTS: One hundred twenty-four patients were entered in this study. Four patients (3.2%) developed septic complications (3 anastomotic leaks and 1 intraabdominal abscess) and were excluded from the analysis. Median age in this population was 68 (range 30-95) years. Mean duration of postoperative ileus was 70+/-28 h (PG) and 99+/-34 h (PS). The type of colectomy, underlying disease, MBP, and early NGT removal failed, in univariate analysis, to correlate with the duration of postoperative ileus. By contrast, time intervals PG and PS were statistically shorter in the group of patients treated by a colorectal surgeon [56+/-23 vs 74+/-28 h (PG); 82+/-26 vs 103+/-35 h (PS), p=0.004], as well as in patients who received systemic opiates for less than 2 days [64+/-27 vs 75+/-28 h (PG), p=0.04; 88+/-32 vs 108+/-33 h (PS), p=001]. CONCLUSION: Restoration of normal intestinal function after elective open colectomy takes 3 (PG) to 4 (PS) days. In our series, specialized training in colorectal surgery has a positive impact on the duration of postoperative ileus. Surgical specialization should be considered an important parameter in future clinical trials aiming to minimize postoperative ileus. PMID- 16267670 TI - Increased nitric oxide (NO) pathway metabolites in the vitreous fluid of patients with rhegmatogenous retinal detachment or diabetic traction retinal detachment. AB - BACKGROUND: Nitric oxide (NO) plays a significant role in physiological and pathological processes in the retina. In the L-arginine-NO pathway, NO synthase (NOS) converts L-arginine to NO and L-citrulline. Increased NO production, mediated by inducible NOS has been implicated in the pathogenesis of various vitreoretinal diseases. In the present study it is hypothesized that in rhegmatogenous retinal detachment (RRD), the production of NO pathway metabolites might be upregulated. METHODS: Using high-pressure liquid chromatography citrulline, arginine and nitrite were measured in vitreous fluid of 93 eyes with RRD, nine eyes with a traction retinal detachment due to proliferative diabetic retinopathy (PDR), and in 49 control samples of vitreous fluid from eyes without retinal detachment. RESULTS: The mean vitreous concentrations of citrulline and arginine were significantly increased in eyes with RRD (9.6+/-4.3 and 97.3+/ 29.2; respectively) or in eyes with a traction retinal detachment (25.8+/-10.3 and 130.7+/-23.7; respectively) as compared to control eyes (7.1+/-3.2 and 75.9+/ 18.1; respectively). The mean level of nitrite was also higher in vitreous fluid of patients with RRD (2.24+/-1.4) or patients with a traction retinal detachment (2.21+/-0.72) than in the controls (2.01+/-0.72), although not significantly so. CONCLUSIONS: We found increased levels of NO pathway metabolites in the vitreous fluid of eyes with retinal detachment, which may reflect a possible role of NO in the pathogenesis of this disease. PMID- 16267671 TI - Ring chromosome 15: characterization by array CGH. AB - Ring chromosome 15 [r(15)] is an uncommon finding with less than 50 patients reported. Precise genotype-phenotype correlations are problematic because of the difficulties in determining the extent of euchromatic loss, the level of mosaicism, and the influence of the timing of ascertainment. We report two discordant examples of r(15) patients. In the first case, prenatal diagnosis of a de novo r(15) was made during the second trimester: mos 46,XX,r(15)(p11.2q26)[32]/45,XX,-15[13]/47,XX,r(15)(p11.2q26)x2[3]/46,XX,dic r(15)(p11.2q26p11.2q26[1]/46,XX[2]. Postnatal follow-up revealed extremely small stature, heart defects, and developmental delay. Patient 2 was a 31-year-old short-statured female who was living independently: 46,XX,r(15)(p11q26). Both cases showed loss of the 15q subtelomeric region by fluorescence in situ hybridization (FISH). To investigate the discordance in phenotypes between the two patients, we undertook array comparative genomic hybridization (array CGH) analyses to more fully characterize the deletions associated with these otherwise structurally indistinguishable r(15) chromosomes from conventional cytogenetic analyses and fluorescence in situ hybridization (FISH) studies. By array CGH, patient 1 showed deletion of multiple contiguous clones predicting an approximately 6 Mb deletion of distal 15q. In contrast, patient 2 showed loss of just the 15q subtelomeric clone and an interstitial clone by array CGH confirming that the severity of the phenotype correlated with the size of the deletion at the molecular level. These cases illustrate the utility of array CGH characterization for determining the size of the associated deletion in ring chromosomes and for facilitating phenotype-genotype correlations. PMID- 16267672 TI - Effects of castration on the expression of brain-derived neurotrophic factor (BDNF) in the vas deferens and male accessory genital glands of the rat. AB - Brain-derived neurotrophic factor (BDNF) is a growth factor belonging to the family of neurotrophins. Although neurotrophins in the male genital organs have been well documented, their role in the biology of these organs is far from clear. In particular, little is known about the influence of sex hormones on neurotrophin expression. In the present study, using immunohistochemistry and enzyme-linked immunosorbent assay (ELISA), we investigated the distribution and tissue concentration of BDNF in the vas deferens and accessory male genital glands in normal and castrated rats. The expression of BDNF mRNA was also investigated. In normal rats, BDNF immunoreactivity was localized in the musculature of the vas deferens and vesicular gland and in the fibromuscular stromal cells of the prostate. In the ventral prostatic lobes, BDNF immunoreactivity was localized in basal, secretory and neuroendocrine epithelial cells. Innervating ganglia and nerves were immunoreactive in all the examined tracts. After castration, BDNF immunoreactivity increased in the musculature of the vesicular gland and in the fibromuscular stromal cells of both dorsal and ventral prostatic lobes. BDNF immunoreactivity also increased in the nerves. ELISA and reverse transcription/real-time polymerase chain reaction confirmed the findings of the immunohistochemical study. In the accessory glands, castration induced an increase of both BDNF tissue concentration and mRNA expression. These results suggest that BDNF is expressed in the internal male genital organs of the rat and that its expression is downregulated by androgen hormones. We hypothesize that the observed BDNF increases are related to the castration-induced regression of the sympathetic nerves. PMID- 16267675 TI - Characterization of a peg-like terminal NOR structure with light microscopy and high-resolution scanning electron microscopy. AB - An atypical peg-like terminal constriction ("peg") on metaphase chromosomes of the plant genus Oziroe could be identified as a nucleolus organizing region (NOR) by detecting 45S rDNA with correlative light microscopy (LM) and scanning electron microscopy (SEM) in situ hybridization (ISH). Using high-resolution 3D analytical SEM, the architecture and DNA distribution of the peg-like NOR were characterized as typical for chromosomes, albeit with significantly smaller chromomeres. ISH procedure was improved for SEM concerning signal localization, labeling efficiency, and structural preservation, allowing 3D SEM analysis of the peg-like NOR structure and rDNA distribution for the first time. It could be shown that implementation of FluoroNanogold markers is an attractive tool that allows efficient immunodection in both LM and SEM. A model is proposed for the peg structure and its mode of condensation. PMID- 16267674 TI - Increased missegregation and chromosome loss with decreasing chromosome size in vertebrate cells. AB - Chromosome engineering has allowed the generation of an extensive and well defined series of linear human X centromere-based minichromosomes, which has been used to investigate the influence of size and structure on chromosome segregation in vertebrate cells. A clear relationship between overall chromosome size and mitotic stability was detected, with decreasing size associated with increasing loss rates. In chicken DT40, the lower size limit for prolonged mitotic stability is approximately 550 kb: at 450 kb, there was a dramatic increase in chromosome loss, while structures of approximately 200 kb could not be recovered. In human HT1080 cells, the size threshold for mitotic stability is approximately 1.6 Mb. Minichromosomes of 0.55-1.0 Mb can be recovered, but display high loss rates. However, all minichromosomes examined exhibited more segregation errors than normal chromosomes in HT1080 cells. This error rate increases with decreased size and correlates with reduced levels of CENP-A and Aurora B. In mouse LA-9 and Indian muntjac FM7 cells, the size requirements for mitotic stability are much greater. In mouse, a human 2.7-Mb minichromosome is rarely able to propagate a kinetochore and behaves acentrically. In Indian muntjac, CENP-C associates with the human minichromosome, but the mitotic apparatus appears unable to handle its segregation. PMID- 16267676 TI - [New therapeutic concepts for vasculitis and collagenosis]. AB - After improvement of the prognosis of the primary systemic vasculitides and systemic lupus erythematosus from a desperate diagnosis with hardly a one year survival after diagnosis to a 5-year-survival-rate of more than 90% actual therapeutic regimes aim at those patients refractory to standard therapeutic regimes, not achieving a remission by standard approaches or having organ damage or contraindications. Furthermore less toxic regimes are looked for with the aim to avoid secondary complications of the standard therapy. New drugs used successfully in rheumatology, transplantation medicine and haematology are used for these purposes in the last years. Recent experiences with Infliximab, Mycophenolate Mofetil, Rituximab und Deoxyspergualin for the treatment of the small vessel vasculitides and systemic lupus erythematosus are reviewed. PMID- 16267677 TI - Tapered terminal ileum conduit for antegrade continence enemas. AB - Antegrade continence enemas can be used to control fecal incontinence caused by neurogenic bowel. Creating a conduit can become a problem when the appendix is unsuitable for use. This paper describes a procedure using a tapered terminal ileum conduit for use with antegrade continence enemas in place of the appendix. This procedure has been attempted and successfully completed on two pediatric patients with good results at 6-month follow-up. The tapered terminal ileum conduit is an option for creating a conduit for antegrade continence enemas in patients without a suitable appendix. PMID- 16267680 TI - Unusual spinal teratoma with an accessory penis on the back. AB - INTRODUCTION: Spinal teratomas are extremely rare tumours, and their association with split cord malformation is even rarer. CASE REPORT: This is a case report of an infant with a large teratoma with a lipomyelomeningocele along with a split cord malformation and with an unusual accessory penis and scrotum on the back over the swelling. This, to our knowledge, is the first such case reported in the English literature. PMID- 16267678 TI - Advances in short bowel syndrome: an updated review. AB - Short bowel syndrome (SBS) continues to be an important clinical problem due to its high mortality and morbidity as well as its devastating socioeconomic effects. The past 3 years have witnessed many advances in the investigation of this condition, with the aim of elucidating the cellular and molecular mechanisms of intestinal adaptation. Such information may provide opportunities to exploit various factors that act as growth agents for the remaining bowel mucosa and may suggest new therapeutic strategies to maintain gut integrity, eliminate dependence on total parenteral nutrition, and avoid the need for intestinal transplantation. This review summarizes current research on SBS over the last few years. PMID- 16267681 TI - Umbilical angiectases as the sole clinical sign of Fabry disease in a 9-year-old boy. PMID- 16267679 TI - T lymphocyte chemotactic chemokines in acute myelogenous leukemia (AML): local release by native human AML blasts and systemic levels of CXCL10 (IP-10), CCL5 (RANTES) and CCL17 (TARC). AB - T cell targeting immunotherapy is now considered in acute myelogenous leukemia (AML), and local recruitment of antileukemic T cells to the AML microcompartment will then be essential. This process is probably influenced by both intravascular as well as extravascular levels of T cell chemotactic chemokines. We observed that native human AML cells usually showed constitutive secretion of the chemotactic chemokines CXCL10 and CCL5, whereas CCL17 was only released for a subset of patients and at relatively low levels. Coculture of AML cells with nonleukemic stromal cells (i.e., fibroblasts, osteoblasts) increased CXCL10 and CCL17 levels whereas CCL5 levels were not altered. However, a wide variation between patients in both CXCL10 and CCL5 levels persisted even in the presence of the stromal cells. Neutralization of CXCL10 and CCL5 inhibited T cell migration in the presence of native human AML cells. Furthermore, serum CCL17 and CXCL10 levels varied between AML patients and were determined by disease status (both chemokines) as well as patient age, chemotherapy and complicating infections (only CCL17). Thus, extravascular as well as intravascular levels of T cell chemotactic chemokines show a considerable variation between patients that may be important for T cell recruitment and the effects of antileukemic T cell reactivity in local AML compartments. PMID- 16267682 TI - Unusual extension of the first branchial cleft anomaly. AB - First branchial cleft is the only branchial structure that persists as the external ear canal, while all other clefts are resorbed. Incomplete obliteration and the degree of closure cause the varied types of first branchial cleft anomalies. They were classified based on the anatomical and histological features. We present an unusual type of first branchial cleft anomaly involving the external auditory canal, the middle ear and the nasopharynx through the eustachian tube. PMID- 16267683 TI - Evaluation of the relation between audiometric and psychometric measures of hearing after tympanoplasty. AB - The objective of this prospective study was to evaluate the relation between audiometric and psychometric measures after tympanoplasty from the perspective of preoperative selection of patients and postoperative assessment of the results of reconstructive middle ear surgery. Hearing (dis)ability was measured by means of pure-tone audiometry and a validated self-assessment questionnaire: the (modified) Amsterdam Inventory of Auditory Disability and Handicap (m)AIAD. Average hearing thresholds and (m)AIAD scores were evaluated for 80 patients, pre and 12 months postoperatively. The average improvement of the air conduction threshold in the operated ear was 5.4 (+/-14.3) dB; the average improvement in the (m)AIAD score was 2.9 points (+/-12.1). Although not very strong, the audiometric improvement and increase in (m)AIAD score are significantly related. After the calculation of postoperatively measured mean scores on the (m)AIAD for different 10-dB intervals of postoperative hearing loss averaged over both ears, an interesting relation between the (m)AIAD score and hearing losses emerges. The (m)AIAD score is almost independent of hearing loss for postoperative hearing levels between 25 and 40 dB. Residual hearing loss has to become less than 25 dB before a smaller hearing loss corresponds with a higher (better) (m)AIAD score. For losses larger than 40 dB the (m)AIAD score clearly decreases with increasing hearing loss. Finally, even small residual hearing losses lead, on average, to (m)AIAD scores that are substantially lower than the score for normally hearing subjects. In general, the patient benefit seems related to the magnitude of improvement in the air-conduction thresholds, rather than to the achievement of a certain threshold level. In addition, even small residual hearing losses (less than 25 dB HL) still lead, on average, to (m)AIAD scores that are substantially lower than the scores for normally hearing subjects. PMID- 16267684 TI - Neck mass as the first presentation of testicular choriocarcinoma. AB - Choriocarcinoma is a highly malignant tumor of the testis and usually occurs as a component of mixed germ cell tumor. The testicular enlargement may be subtle, and the patient may present with metastasis, the neck being one of its least reported locations. A case of testicular choriocarcinoma in a 24-year-old man who presented for the first time with metastatic neck mass is reported here. This presentation, although very rare, can be a great diagnostic and therapeutic challenge and should be considered in the differential diagnosis of cervical masses occurring in young males. PMID- 16267685 TI - Postural stability in patients with different durations of benign paroxysmal positional vertigo. AB - Postural stability maintenance was investigated in patients with "idiopathic" benign paroxysmal positional vertigo of the posterior semicircular canal (BPPV PSC) and compared to healthy subjects. To measure the postural stability during a quiet upright stance, we used static posturography in two conditions: with open and with closed eyes. The effect of the repositioning Epley's maneuver on the recovery of postural stability in patients with different durations of BPPV-PSC less than 60 days after the first attack of positioning vertigo (group I) and more than 60 days (group II) was examined. The investigation was made 1 h after the positive Dix-Hallpike test and 7 days after treatment with the Epley maneuver. "Sway velocity" (SV) and "relative power spectrum" (RPS) of the stabilograms were calculated to evaluate the postural stability for each subject and each experimental condition. We found a pronounced spectral density peak in the frequency range of 1.0-2.0 Hz and smaller spectral density in the range of 0.15-0.5 Hz compared to that in healthy subjects. Our results showed that the postural maintenance in BPPV-PSC patients depended on the disease duration. Patients with a duration of BPPV symptoms less than 60 days after the first attack demonstrated a high dependence on the visual input for postural stability. One week after the Epley maneuver, differences in the recovery of postural control in both groups of patients were also found. We assume that the disturbed otolith function together with the impaired dynamics of the semicircular canal generate a particular pattern of postural maintenance. The different degree of restoration of postural stability 1 week after the EM treatment in BPPV-PSC patients with different durations of the disease gives us reason to believe that after removing the otoconia from the semicircular canal, some stimulation of other sensory subsystems and adaptation mechanisms occur that lead to a new pattern of postural maintenance. PMID- 16267686 TI - Chloroacetaldehyde- and acrolein-induced death of human proximal tubule cells. AB - Ifosfamide (ifo) is a commonly used drug in chemotherapy. It is metabolized to acrolein (acro) and chloroacetaldehyde (CAA), which are thought to be responsible for renal side effects. We studied the effects of ifo and cyclophosphamide (cyclo) as well as their metabolites, acro and CAA, on cellular protein content, necrosis, apoptosis and cytosolic calcium concentration using a human proximal tubule cell line. The protein content decreased during acro or CAA administration (15 to 300 micromol/l), but not during ifo or cyclo exposure over a time period of up to 72 h. Mild apoptosis was induced only by high acro (150, 300 micromol/l) and low CAA concentrations (15, 75 micromol/l) and only in a narrow time window (24 h). Necrosis was increased after exposure to acro or CAA at all concentrations. CAA was more potent than acro. Ifo and cyclo did not induce necrosis or apoptosis. Glutathione abolished CAA-induced cell death. Cytosolic calcium concentrations increased after acro or CAA administration and showed an oscillating pattern. Cytosolic Ca(2+) chelation did not prevent necrosis. We conclude that neither ifo nor cyclo induce cell damage, but that their metabolites acro and CAA induce cell death. This cell death occurs mainly by necrosis and not by apoptosis. PMID- 16267688 TI - Protein structure prediction by tempering spatial constraints. AB - The probability to predict correctly a protein structure can be enhanced through introduction of spatial constraints - either from NMR experiments or from homologous structures. However, the additional constraints lead often to new local energy minima and worse sampling efficiency in simulations. In this work, we present a new parallel tempering variant that alleviates the energy barriers resulting from spatial constraints and therefore yields to an enhanced sampling in structure prediction simulations. PMID- 16267687 TI - [Hypereosinophilic dermatitis. An overlooked diagnosis?]. AB - BACKGROUND: The idiopathic hypereosinophilic syndrome is a rare systemic disease characterized by blood and tissue eosinophilia of unknown etiology, in which multiple organs may be affected. In hypereosinophilic dermatitis the only affected organ besides the blood is the skin. PATIENTS: We present a series of seven patients with hypereosinophilic dermatitis who were treated in our hospital between 2002 and 2003. RESULTS: All patients initially showed characteristic, therapy-resistant, polymorphic skin lesions, presenting with a combination of erythematous, pruritic and urticarial papules and plaques. All had blood eosinophilia without evidence of allergic, parasitic or other causes. The histology showed tissue eosinophilia only in half of the cases; the other histological findings were non-specific. We observed a good response to therapy with systemic corticosteroids, dapsone and light therapy, applied as UVA-1 irradiation or as shower photochemotherapy. CONCLUSIONS: The diagnosis "hypereosinophilic dermatitis" should be based primarily on the characteristic clinical picture together with "idiopathic" peripheral eosinophilia, whereas the histological findings are not always indicative. Because of the multiplicity of possible differential diagnoses and the often non-revealing histology, we assume that the diagnosis "hypereosinophilic dermatitis" is often overlooked. PMID- 16267690 TI - Why relevant chemical information cannot be exchanged without disclosing structures. AB - Both society and industry are interested in increasing the safety of pharmaceuticals. Potentially dangerous compounds could be filtered out at early stages of R&D by computer prediction of biological activity and ADMET characteristics. Accuracy of such predictions strongly depends on the quality & quantity of information contained in a training set. Suggestion that some relevant chemical information can be added to such training sets without disclosing chemical structures was generated at the recent ACS Symposium. We presented arguments that such safety exchange of relevant chemical information is impossible. Any relevant information about chemical structures can be used for search of either a particular compound itself or its close analogues. Risk of identifying such structures is enough to prevent pharma industry from relevant chemical information exchange. PMID- 16267691 TI - Surrogate data--a secure way to share corporate data. AB - The privacy of chemical structure is of paramount importance for the industrial sector, in particular for the pharmaceutical industry. At the same time, companies handle large amounts of physico-chemical and biological data that could be shared in order to improve our molecular understanding of pharmacokinetic and toxicological properties, which could lead to improved predictivity and shorten the development time for drugs, in particular in the early phases of drug discovery. The current study provides some theoretical limits on the information required to produce reverse engineering of molecules from generated descriptors and demonstrates that the information content of molecules can be as low as less than one bit per atom. Thus theoretically just one descriptor can be used to completely disclose the molecular structure. Instead of sharing descriptors, we propose to share surrogate data. The sharing of surrogate data is nothing else but sharing of reliably predicted molecules. The use of surrogate data can provide the same information as the original set. We consider the practical application of this idea to predict lipophilicity of chemical compounds and we demonstrate that surrogate and real (original) data provides similar prediction ability. Thus, our proposed strategy makes it possible not only to share descriptors, but also complete collections of surrogate molecules without the danger of disclosing the underlying molecular structures. PMID- 16267692 TI - Prediction of plasma protein binding of drugs using Kier-Hall valence connectivity indices and 4D-fingerprint molecular similarity analyses. AB - A 115 compound dataset for HSA binding is divided into the training set and the test set based on molecular similarity and cluster analyses. Both Kier-Hall valence connectivity indices and 4D-fingerprint similarity measures were applied to this dataset. Four different predictive schemes (SM, SA, SR, SC) were applied to the test set based on the similarity measures of each compound to the compounds in the training set. The first algorithmic scheme (SM) predicts the binding affinity of a test compound using only the most similar training set compound's binding affinity. This scheme has relatively poor predictivity based both on Kier-Hall valence connectivity indices similarity measures and 4D fingerprints similarity analyses. The other three algorithmic schemes (SM SR, SC), which assign a weighting coefficient to each of the top-ten most similar training set compounds, have reasonable predictivity of a test set. The algorithmic scheme which categorizes the most similar compounds into different weighted clusters predicts the test set best. The 4D-fingerprints provide 36 different individual IPE/IPE type molecular similarity measures. This study supports that some types of similarity measures are highly similar to one another for this dataset. Both the Kier-Hall valence connectivity indices similarity measures and the 4D-fingerprints have nearly same predictivity for this particular dataset. PMID- 16267689 TI - ENPDA: an evolutionary structure-based de novo peptide design algorithm. AB - One of the goals of computational chemists is to automate the de novo design of bioactive molecules. Despite significant advances in computational approaches to ligand design and binding energy evaluation, novel procedures for ligand design are required. Evolutionary computation provides a new approach to this design endeavor. We propose an evolutionary tool for de novo peptide design, based on the evaluation of energies for peptide binding to a user-defined protein surface patch. Special emphasis has been placed on the evaluation of the proposed peptides, leading to two different evaluation heuristics. The software developed was successfully tested on the design of ligands for the proteins prolyl oligopeptidase, p53, and DNA gyrase. PMID- 16267693 TI - Secure analysis of distributed chemical databases without data integration. AB - We present a method for performing statistically valid linear regressions on the union of distributed chemical databases that preserves confidentiality of those databases. The method employs secure multi-party computation to share local sufficient statistics necessary to compute least squares estimators of regression coefficients, error variances and other quantities of interest. We illustrate our method with an example containing four companies' rather different databases. PMID- 16267694 TI - Reverse engineering chemical structures from molecular descriptors: how many solutions? AB - Physical, chemical and biological properties are the ultimate information of interest for chemical compounds. Molecular descriptors that map structural information to activities and properties are obvious candidates for information sharing. In this paper, we consider the feasibility of using molecular descriptors to safely exchange chemical information in such a way that the original chemical structures cannot be reverse engineered. To investigate the safety of sharing such descriptors, we compute the degeneracy (the number of structure matching a descriptor value) of several 2D descriptors, and use various methods to search for and reverse engineer structures. We examine degeneracy in the entire chemical space taking descriptors values from the alkane isomer series and the PubChem database. We further use a stochastic search to retrieve structures matching specific topological index values. Finally, we investigate the safety of exchanging of fragmental descriptors using deterministic enumeration. PMID- 16267696 TI - Pulmonary complications in patients with hematological disorders: pathobiological bases and practical approach. AB - Pulmonary complications occur in up to 40 to 60% of patients with hematological disorders during the disease course and considerably influence morbidity and mortality. The main factors making the lung a clinically significant targeted organ in these patients may be summarized as follows. In the lung parenchyma a variety of inflammatory cells whose precursors are in the bone marrow pass through, park in, proliferate, and release microbicidal and cytohistotoxic substances. Constitutive parenchymal lung cells (bronchiolar and alveolar epithelial cells, endothelial cells, "interstitial" cells) may be a distinctive target for toxic substances or may have an important part in the inflammatory/reactive and reparative processes after an injury event. Pathogenic agents are allowed to reach the lung very easily through either or both the airways and the vascular bed and accumulate there in large amounts. Inflammatory/immunologic reactions may be particularly weak or, on the contrary strong, in the lungs either spontaneously or due to toxic action of drugs and radiation or to the immunodeficiency induced by hematological disorders, and finally to the presence of immunomodulatory viruses. The distinctive anatomical structure and function of the lung parenchyma (interactions between air spaces and capillary bed-gas exchange units) may render localized parenchymal damage clinically relevant. Allogeneic reactions may be overexpressed in the lung or the kinetics of the developing of graft versus host disease (GVHD)-related lung injury may be markedly different from the kinetics of GVHD in other organs. Hematological disorders may harbor in lung parenchymal structures at the onset (i.e., lympho-/myeloproliferative disorders primary in the lung) or during the disease course. Genetic predisposition, although probably involved, is not yet well understood. This article reviews the pathobiological bases of lung injury occurring in subjects with hematological disorders and suggests a practical diagnostic approach to these pulmonary complications. PMID- 16267697 TI - Pulmonary infections complicating hematological disorders. AB - Pulmonary infections are second in importance only to septicemia as a cause of infectious morbidity and mortality in patients with hematological disorders. The differential diagnosis of the pneumonitis syndrome includes not only infection but also a multitude of noninfectious causes. In addition, the diagnosis may be difficult, owing to the subtlety of the clinical signs as a consequence of the impaired inflammatory response. Radiographic findings are often nonspecific, and invasive procedures and microbiological exams are required to establish the cause of pulmonary disease and to choose a specific therapy. However, invasive diagnostic procedures are often precluded by the poor general conditions and (particularly in acute leukemia patients) by concurrent thrombocytopenia. The approach to all infectious complications, including those of the lower respiratory tract, in immunocompromised patients with hematological diseases, is based on aggressive prevention strategies and the empirical administration of broad-spectrum antimicrobials eventually followed by a clinically or microbiologically guided treatment modification. With regard to the antimicrobial treatment, given the variety of infectious and noninfectious causes of pulmonary infiltrates in patients with hematological diseases, the diversity of the underlying immunocompromised state, and the spectrum of clinical findings, no single general therapeutic algorithm can be applied. PMID- 16267698 TI - Drug-induced respiratory disease in patients with hematological diseases. AB - Patients with hematological malignancies or recipients of hematopoietic stem cell transplants may develop myriad pulmonary manifestations, as a complication of either the disease or the diverse agents used to treat the disease. Clinical, radiographic, and physiological features of drug-induced and radiation-induced pulmonary injury are often difficult to distinguish from other causes of pulmonary infiltrates (e.g., infections, pulmonary edema, alveolar hemorrhage, etc.). Fiberoptic bronchoscopy with bronchoalveolar lavage (BAL) is essential to exclude infectious etiologies. In some cases, surgical lung biopsies are required to establish a specific etiological diagnosis. This review discusses the myriad causes of lung injury/toxicity that may afflict patients with hematological malignancies or transplant recipients, and presents diagnostic and therapeutic approaches. PMID- 16267699 TI - Graft versus host-associated pulmonary disease and other idiopathic pulmonary complications after hematopoietic stem cell transplant. AB - Noninfectious conditions are now the major pulmonary causes of morbidity and mortality after hematopoietic stem cell transplantation (HSCT). The idiopathic pneumonia syndrome (IPS) remains one of the more common and serious pulmonary complications within months after transplantation. Epidemiological data suggest that, although graft versus host disease (GVHD) reactions may play an etiological role, the major contributing factor is conditioning-related toxicity. Among lung conditions that are more closely associated with GVHD are both obliterative bronchiolitis (usually occurring months to years after HSCT) and bronchiolitis obliterans organizing pneumonia (BOOP). The former is an inexorably progressive condition, whereas BOOP behaves similarly to idiopathic BOOP seen in other populations. Also addressed in this article are engraftment syndrome, diffuse alveolar hemorrhage, and pulmonary veno-occlusive disease. Knowledge of these complications is now a part of the contemporary practice of pulmonary medicine, no longer isolated to the transplant pulmonologist. PMID- 16267700 TI - Lymphoproliferative lung disorders. AB - Although the lung is frequently involved by disseminated lymphoma, isolated pulmonary lymphoma is rare, accounting for less than 1% of all extranodal localized disease. Three broad categories of lymphoma of the lung require recognition: in rare instances, large B cell type lymphoma can present primarily in the lung; a second variant is by T cell lymphoma presenting as an angiocentric process. However, the most common histologic subtype is represented by low-grade mucosa-associated lymphoid tissue (MALT) lymphoma, often in the past considered as a pseudotumor because of its long indolent natural history. Common presenting features include cough, dyspnea, pain, fever, recurrent infections, hemoptysis, or an asymptomatic finding on routine chest radiograph. Precise pathological diagnosis and molecular characterization are required in all cases, following World Health Organization classification criteria. Radiological features include pulmonary consolidation, solid pulmonary opacities, hilar adenopathy, or pleural effusion. Principles of treatment vary with the different histology. PMID- 16267702 TI - Myeloid leukemias and lung involvement. AB - Myeloid leukemias are clonal malignancies characterized by the presence of increased numbers of immature myeloid cells in the marrow and peripheral blood. Pulmonary involvement by myeloid leukemia is relatively uncommon and seen mainly in patients with severe disease. The most common form of pulmonary involvement consists of leukemic infiltration along the lymphatics in the peribronchovascular, septal, and pleural interstitial tissue. Less common manifestations include myeloid sarcoma, leukostasis, leukemic cell lysis pneumopathy, and hyperleukocytic reaction. The radiological manifestations of pulmonary leukemic cell infiltration and leukostasis consist mainly of bilateral thickening of the peribronchovascular interstitium and interlobular septa, a pattern that resembles that of interstitial pulmonary edema. The radiological manifestations of leukemic cell lysis pneumopathy and hyperleukocytic reaction consist of symmetric bilateral areas of consolidation. This manuscript reviews the histological and radiological intrathoracic manifestations of myelogenous leukemias. PMID- 16267701 TI - Pulmonary amyloidosis in hematological disorders. AB - Amyloidosis is defined by tissue deposits of amyloid, a proteic substance with a characteristic spatial structure of beta-sheet fibrils assembled into bundles. This structure results in specific staining with Congo red dye, with green birefringence under polarized light microscope. AL amyloid forms from amyloidogenic immunoglobulins produced by clonal plasma cells. Pulmonary amyloidosis may be either part of systemic amyloidosis (primary amyloidosis or associated with myeloma) or organ-limited. Pulmonary interstitial amyloidosis in systemic amyloidosis is rarely symptomatic unless amyloid deposits severely affect gas exchange alveolar structures, thus resulting in serious respiratory impairment. Localized parenchymal involvement may present as nodular amyloidosis or as amyloid deposits in the vicinity of pulmonary lymphomas. Tracheobronchial amyloidosis, which is not associated with evident clonal proliferation, results in airway stenoses necessitating iterative laser treatment. Treatment of systemic amyloidosis aims at reducing the clonal cell populations producing amyloidogenic immunoglobulins, using high-dose chemotherapy followed by autologous stem cell transplantation in carefully selected patients. Its efficiency in treating diffuse pulmonary amyloidosis has not been established. PMID- 16267703 TI - Secondary lung tumors in hematological patients. AB - Advances made in the field of chemotherapy and radiotherapy have considerably increased the survival of patients with Hodgkin's disease (HD), non-Hodgkin's lymphoma (NHL), and chronic lymphocytic leukemia (CLL). Unfortunately, these antiblastic therapies have also increased the risk of late complications such as second tumors, especially second lung cancers. Although the role of ionizing radiations in carcinogenesis is now clear, less is known about the damage caused by chemotherapy, immunodeficiency induced by drugs or hematological pathologies, and cigarette smoking. In HD, the relative risk (RR) of second lung cancer increases considerably in relation to the dose of ionizing radiation given to the patient, with an RR of 9.6 when more than 9 Gy are administered. Some studies have reported a significantly higher risk of second lung cancers in smokers compared with nonsmokers ( p = .03). The role of chemotherapy in the development of second lung cancers has yet to be determined. Although some authors correlate a greater risk with an increased number of chemotherapy cycles, others maintain that chemotherapy increases the risk of second lung cancer only if associated with cigarette smoking. Even less is known about the correlation between NHL and second lung cancer. Although the RR is higher in long-term NHL survivors than in healthy individuals (RR = 1.36), the heterogeneity of histotype and treatment does not permit us to confirm a correlation with chemotherapy and smoking. Conversely, in CLL, the development of second lung cancer appears to be linked to the immunodeficiency that accompanies this hematological malignancy. This is confirmed by the identical RR (1.66) for CLL patients subjected to chemotherapy and for those who have only follow-up. PMID- 16267704 TI - Lungs in hemoglobinopathies, erythrocyte disorders, and hemorrhagic diatheses. AB - The erythropoietic system plays a major role in tissue oxygenation because the erythrocytes are the primary carriers of oxygen in the form of oxyhemoglobin. Therefore, clinical entities such as abnormal hemoglobins, polycythemia, anemia, and significant changes in blood volume frequently produce alterations in various respiratory functions. The pulmonary manifestations can vary from mild respiratory illness to life-threatening emergencies with high mortality rates. Among the hemoglobinopathies, sickle cell disease is clinically the most important and commonly associated with serious pulmonary consequences, including acute chest syndrome, pneumonia, infarction due to in situ thrombosis, bone marrow fat embolism of pulmonary vasculature, bone marrow infarction, pulmonary hypertension, and other abnormalities. Hemoglobinopathies with high and low affinity for oxygen and other abnormal hemoglobinopathies occasionally cause clinically significant respiratory complications by interfering with normal tissue oxygenation. Acquired methemoglobinemia can cause alarming cyanosis and medical emergency. Erythrocyte disorders are associated with pulmonary complications, including pulmonary hypertension, alveolar fibrosis, and pulmonary dysfunction. Coagulation disorders, both the inherited and acquired types, have the potential to affect the respiratory system in the form of hemorrhage from the airways, lung parenchyma, or pulmonary hypertension. The following paragraphs describe the common pulmonary complications and manifestations associated with hemoglobinopathies, erythrocyte disorders, and coagulation abnormalities. PMID- 16267705 TI - [Nobel prize in 2005 for H. Pylori and for gastroenterology]. PMID- 16267706 TI - [Influence of adjuvant radio-chemotherapy for rectal cancer on quality of life]. AB - INTRODUCTION: Modern therapy for rectal cancer is associated with functional disorders. Dysfunction as a consequence of surgery has to be distinguished from disorders caused by postoperative therapy. Therefore we have compared the long term functional results of patients who received postoperative radio-chemotherapy or no therapy in conjunction with low anterior resection of the rectum. PATIENTS AND METHOD: From 1997 to 2002, a total of 32 patients (16 males and 16 females) after low anterior rectal resection and postoperative radio-chemotherapy or surgical therapy alone was compared using standardized and validated instruments (Short-Form-36-Health-Survey, EORTC QLQ-C30, QLQ-CR 38 and ASCRS fecal incontinence questionnaire) in a matched-pair analysis (age, gender and time of surgery). Mean age was 61.8 (62.1) years and mean follow-up was 4 (3.8) years. RESULTS: Two out of the 40 examined parameters differed significantly. There were no significant differences in Short-Form-36-Health-Survey and EORTC QLQ-C30 scales between both groups. The QLQ-CR38 scale sexual enjoyment differed significantly, whereas future perspectives, sexual functioning, micturition problems, symptoms in the area of the gastrointestinal tract, weight loss, defecation problems, male and female sexual problems did not differ significantly. The scales Lifestyle, Coping/Behavior and Depression/Self Perception of the ASCRS fecal incontinence questionnaire also did not differ significantly. The difference for embarrassment was significant. DISCUSSION: No differences in quality of life after postoperative radio-chemotherapy or no postoperative therapy in conjunction with low anterior rectal resection can be found. There are, however, tendencies that postoperative radio-chemotherapy has more adverse effects on continence and sexual function than low anterior rectal resection alone. PMID- 16267707 TI - Contrast-enhanced endoscopic ultrasound with low mechanical index: a new technique. AB - AIM: Recent advances in technology have supported the development of new endoscopic ultrasound systems making it possible to use low MI contrast-enhanced imaging techniques (wide band harmonic imaging done with endoscopic ultrasound is currently at a preliminary stage). We now report on the first use of contrast enhanced, low mechanical index, real-time endoscopic ultrasound (CELMI-EUS) in six patients using prototype technology. MATERIALS AND METHODS: CELMI-EUS was performed using an electronic echo-endoscope HITACHI/Pentax EG-3830UT and adapted dynamic contrast harmonic wide-band pulsed inversion software with low mechanical index (MI = 0.09 - 0.25) before and up to 180 seconds after injection of SonoVue (4.8 mL) in six patients. RESULTS: Adequate visualisation of the arterial and portal venous phases was achieved in all patients. The pancreas and liver were studied thereafter. In contrast to the satisfactory visualisation of these vessels, enhancement of the left liver lobe was sufficient only in 4 patients. In the remaining 2 patients with liver cirrhosis, the enhancement was less pronounced in contrast to the strong enhancement of the hepatic artery and portal vein. CONCLUSION: Recent advances in technology have supported the development of new echo-endoscopic systems making it possible to use real-time, low mechanical index, contrast-enhanced imaging techniques with endoscopic ultrasound. We have preliminarily shown that arterial, portal venous and parenchymal contrast enhancement is possible. PMID- 16267708 TI - [Splenosis--important differential diagnosis in splenectomized patients presenting with abdominal masses of unknown origin]. AB - A 40-year-old female patient was admitted for work-up of multiple abdominal masses. The lymphoma-mimicking tumors were detected accidentally during an ultrasound course. The past medical history was unremarkable besides a status post-traumatic splenic rupture and splenectomy. The patient was asymptomatic, especially there were no complaints of fever, night sweats or weight loss. Laboratory tests did not show pathological results. Ultrasound of the abdomen revealed multiple hypoechoic mesenterial and peritoneal enlarged tumors as well as a subhepatic mass (30 x 20 mm). Transmission computed tomography (CT) showed a normal chest, excluded abnormal thoracal masses and confirmed the multiple abdominal nodules. Microparticles were trapped only by tissue with phagocytosis function as cells of the reticulohistiocytary system in liver and spleen. Uptake of (99 m)Tc-labeled microparticles is specific for splenic tissue. All abdominal masses were detectable by single photon emission computed tomography (SPECT) after intravenous administration of this radiotracer. Ultrasound-guided biopsy proved the presence of spleen tissue with follicular hyperplasia. In conclusion, we report a case of post-traumatic splenosis. In 16 - 67 % of patients who experienced traumatic splenic rupture autotransplanted spleen tissue can be detected. Splenosis therefore is an important differential diagnosis of abdominal masses in splenectomized patients. PMID- 16267709 TI - [Enteral nutrition: drug administration via feeding tube]. AB - Enteral nutrition support via a feeding tube is a preferred and broadly applied way of artificial nutrition in patients who cannot take up orally an adequate amount of nutrients. These patients often need simultaneous drug therapy as well. Thus, there is a high risk of drug-nutrient interactions. Although enteral nutrition is commonly used there is a lack of awareness and knowledge about the appropriate handling and drug administration via the feeding tube. On the one hand, drug-nutrient interactions can lead to clogging of the tube, on the other hand, the change in bioavailability of the drug can have a direct effect on the therapeutic effort. To optimise safety and efficacy of drug therapy in patients with feeding tubes, some basic rules have been set up. PMID- 16267711 TI - [ASCO-Update 2005--Highlights of the 41st Meeting of the American Society of Clinical Oncology/ASCO 2005]. AB - Currently, the treatment of gastrointestinal cancers is rapidly changing due to the implementation of novel chemotherapeutic agents as well as the introduction of targeted therapies into treatment protocols. The following review provides an overview of the most important clinical trials in esophageal, gastric, colorectal, pancreatic and hepatobiliary cancer that were presented at the annual meeting of the American Society of Clinical Oncology. PMID- 16267710 TI - [Diagnosis of celiac disease]. AB - In spite of modern diagnostic possibilities, the diagnosis of celiac disease is still challenging for the physician. This is due to the great variability of the clinical presentation. Nowadays, the classical symptoms like diarrhea, weight loss and abdominal pain are seen less often. It has become evident that celiac disease is not merely a disease of the intestine but of the entire organism. Furthermore, extraintestinal manifestations can present without any gastrointestinal symptoms. It is likely that in many cases the immune system and not nutrient deficiencies play a major role. In addition, the diagnostic tests are not always unequivocal. There is a great variability of the small intestinal changes which are sometimes in contradiction to the antibody results. Since celiac disease implies a lifelong gluten-free diet, a firm diagnosis should be obtained. Thus, one should not rely on a single test but should rather consider serology, histology and clinical response altogether. PMID- 16267712 TI - Diarrhea at the summit. PMID- 16267713 TI - Global impact of diarrheal diseases that are sampled by travelers: the rest of the hippopotamus. AB - Travelers who experience diarrhea (i.e., "turista") are exposed to the same pathogens and illnesses that pose some of the greatest threats to life and development among malnourished children in developing areas around the world, where inadequate water and poor sanitation remain. This article focuses on new findings about the impact, diagnosis, and control of these illnesses and the genetic predispositions of persons who acquire them. Despite the reductions in mortality due to dehydrating diarrhea, the morbidity associated with diarrheal illnesses continues unabated. Furthermore, we increasingly recognize the lasting detrimental effects of enteric infections that occur during early childhood on later physical and cognitive development and, in patients with acquired immunodeficiency syndrome, on the absorption of antiretroviral drugs. Genetic predispositions to inflammation and potential protection associated with such alleles as ApoE4, which are not suspected of being involved in diarrhea, remind us of how much we have to learn about the effect and interactions of enteric tropical infectious diseases with regard to our host genome. New diagnostic methods hold promise for improved recognition and, hopefully, control of enteric infections worldwide. PMID- 16267714 TI - Looking for evidence that personal hygiene precautions prevent traveler's diarrhea. AB - In the 50 years during which traveler's diarrhea has been studied, it has always been assumed that personal hygiene precautions can prevent or reduce the likelihood of developing traveler's diarrhea. However, 7 of 8 studies that specifically addressed this issue showed no correlation between the types of food selected and the risk of acquiring traveler's diarrhea. The eighth study showed a correlation between a few dietary mistakes and a decreased risk of acquiring traveler's diarrhea. A further increase in the number of dietary mistakes, however, did not continue to increase the risk of acquiring traveler's diarrhea. Personal hygiene precautions, when performed under the direct supervision of an expatriate operating his or her own kitchen, can prevent traveler's diarrhea, but poor restaurant hygiene in most developing countries continues to create an insurmountable risk of acquiring traveler's diarrhea. PMID- 16267715 TI - Epidemiology of traveler's diarrhea. AB - Among travelers from developed countries who visit developing countries, >60% may experience traveler's diarrhea, accounting for 40,000 travelers daily or >15 million travelers annually. Traveler's diarrhea is often accompanied by other symptoms, most often abdominal cramps. Although the spontaneous cure occurs after a mean of 4 days, a few patients have symptoms for weeks, and it is increasingly noted that some patients may later develop irritable bowel syndrome. Traveler's diarrhea is life threatening only exceptionally, but it frequently it leads to incapacitation. Both host factors (e.g., age, behavior, nationality, and genetic factors) and environmental factors (primarily the selected destination and hotel) play an important role in risk for traveler's diarrhea. PMID- 16267716 TI - Diarrhea in nontravelers: risk and etiology. AB - Acute diarrheal illnesses in nontravelers are common and represent a significant health and economic burden in the United States and other developed countries. The likelihood of experiencing diarrhea is increased many fold during travel to developing countries. Extensive overlap exists in the pathogens that cause diarrhea in travelers and nontravelers, although proportions differ and show variation by geographic area and by season, and they change over time. Rates of infection are highest in infants and young children, in whom viral pathogens predominate. Person-to-person transmission may account for more than one-half of cases. In contrast, in many studies, bacterial infections predominate in travelers, who often acquire infection from contaminated food and water. Because of the globalization of the food supply, clinicians in developed countries should expect to continue to see sporadic cases and outbreaks of diarrhea caused by unusual pathogens, such as Cyclospora species. PMID- 16267717 TI - Traveler's diarrhea in the pediatric population: etiology and impact. AB - Children who travel are at risk of developing the same, well-known illnesses that affect adult travelers. The etiology, treatment, and actual risk of these illnesses are not well defined in children. The limited amount of data available forces clinicians to extrapolate appropriate treatment for children. Recommendations for children have not been standardized. The role and judicious use of antimicrobials in the treatment of pediatric traveler's diarrhea have evolved during recent decades and will be reviewed here. The past 2 decades have seen an unprecedented, sustained growth in travel. The epidemiology related to travel medicine has evolved to describe travelers and their illnesses more specifically. The development of the field of travel medicine has led to greater understanding and improved, targeted treatment of travel-related illnesses. Most of the data available today describe traveler's diarrhea in adults. Children traveling to distant destinations from their home country have not been well studied, yet treatment parameters exist in practice and will be reviewed here. PMID- 16267718 TI - Prophylactic antimicrobials for traveler's diarrhea: an early history. AB - The prevention of traveler's diarrhea by use of pharmacologic agents has been of great interest since the early 1960s. It was not until the discovery of enterotoxigenic Escherichia coli as the dominant causative organism of travelers' diarrhea, however, that rational antimicrobial therapy could be studied. Selected antimicrobials have proven to be highly efficacious, but, by consensus, are not widely used. PMID- 16267719 TI - Nonantimicrobial agents in the prevention and treatment of traveler's diarrhea. AB - Among the nonantimicrobial agents that are available and useful for the prevention of traveler's diarrhea are bismuth subsalicylate-containing preparations, which can provide a rate of protection of up to 65% when taken 4 times daily. In one study, the probiotic Lactobacillus GG was found to provide 49% protection against traveler's diarrhea, but results with this agent and other probiotics have been highly variable and geographically inconsistent. Tannin albuminate plus ethacridine lactate provided 36% protection, but it is not widely available. Among the nonantimicrobial agents that are available and useful for the treatment of traveler's diarrhea are bismuth subsalicylate-containing preparations, which reduce the passage of loose stools by 16%-18%. The antisecretory and antimotility agent loperamide reduces the passage of loose stools by approximately 50% and has been especially useful, in combination with antimicrobial agents, in reducing the total duration of posttreatment diarrhea to a matter of hours. PMID- 16267720 TI - Poorly absorbed antibiotics for the treatment of traveler's diarrhea. AB - This article describes the use of poorly absorbed antibiotics for the treatment of traveler's diarrhea. Poorly absorbed oral antibiotics can deliver high concentrations of drug to the site of enteric infection with minimal risk of systemic adverse effects, toxicity, and drug interactions. Compared with systemically absorbed oral antibiotics, poorly absorbed oral antibiotics may be less associated with the pressure that leads to the development of bacterial resistance, because they do not affect bacteria outside the gastrointestinal tract. In clinical studies, poorly absorbed oral antibiotics, including aztreonam, bicozamycin, and rifaximin, were more effective than and as well tolerated as placebo; in particular, rifaximin was as effective as oral ciprofloxacin in reducing the duration of illness in traveler's diarrhea. More research is warranted to delineate the effects of poorly absorbed antibiotics in invasive infection and to assess the potential for the development of bacterial resistance. PMID- 16267722 TI - Sequelae of traveler's diarrhea: focus on postinfectious irritable bowel syndrome. AB - Traveler's diarrhea is usually an acute, self-limited illness; however, in some patients, enteric symptoms can persist for weeks, months, or years. It has been estimated that up to 3% of patients with traveler's diarrhea have symptoms for >30 days. The differential diagnosis includes persistent infection, coinfection, temporary postinfection phenomena, or malabsorptive syndromes. Once these possibilities are excluded, and if symptoms persist, a diagnosis of postinfectious irritable bowel syndrome (PI-IBS) becomes more likely. PI-IBS has recently become a topic of considerable clinical and investigative interest, because evidence validating it as a diagnosis and elucidating its pathophysiological mechanisms has accumulated. Epidemiological evidence suggests that PI-IBS is a relatively common sequela of acute gastroenteritis. Experimental evidence suggests that chronic inflammation following acute bacterial infection has a pathophysiological role in the development of PI-IBS. A fuller understanding of these pathophysiological mechanisms will lead to a more directed therapeutic approach and, perhaps, a reevaluation of prophylaxis for traveler's diarrhea as a means of primary prevention of PI-IBS. PMID- 16267721 TI - Antibacterial chemoprophylaxis in the prevention of traveler's diarrhea: evaluation of poorly absorbed oral rifaximin. AB - The use of antibacterial drugs was first shown to effectively reduce the occurrence of traveler's diarrhea nearly 50 years ago. The approach was not encouraged for general use by a Consensus Development Conference in 1985 because of concerns about adverse effects of the drugs and the possible development of resistance against systemically absorbed drugs. When therapy with poorly absorbed rifaximin was shown to be as effective as therapy with systemically absorbed drugs in shortening the duration of traveler's diarrhea, without the development of resistant coliform flora, the use of rifaximin for the prevention of traveler's diarrhea was studied. In the present study, rifaximin provided 72% protection against the development of diarrhea and 77% protection against active or treated diarrhea during 2 weeks of drug administration to United States students in Mexico. Rifaximin offers a potentially useful approach for preventing traveler's diarrhea. Potential areas of future study include use of the drug to prevent diarrhea due to mucosally invasive bacteria, including ciprofloxacin resistant Campylobacter species, and to reduce the occurrence of postinfectious irritable bowel syndrome. PMID- 16267723 TI - Effective combination therapy using interferon-gamma and interleukin-2 for disseminated Mycobacterium avium complex infection in a pediatric patient with AIDS. AB - A 5-year-old girl presented with disseminated Mycobacterium avium complex infection during advanced human immunodeficiency virus infection. Interferon gamma or interleukin-2 monotherapy showed only limited effects. Use of a combination of interferon-gamma and interferon-2 resulted in a remarkable improvement in the patient's condition, accompanied by an increase in circulating CD4+ T cells. PMID- 16267724 TI - Invasive infections with Haemophilus influenzae serotype a containing an IS1016 bexA partial deletion: possible association with virulence. AB - BACKGROUND: Recent reports of invasive Haemophilus influenzae non-b capsular serotypes in the era since development of conjugate vaccines have prompted concern about serotype replacement. Unusual clusters of invasive infection due to H. influenzae serotype a with clinical features that resemble those of infection due to H. influenzae serotype b have been described. A unique feature often associated with more-virulent H. influenzae serotype a isolates is the IS1016 bexA partial deletion, which was previously identified in the capsule locus of H. influenzae serotype b strains. We report the clinical, epidemiologic, and molecular genetic features of 2 cases of severe disease caused by H. influenzae serotype a. METHODS: Invasive H. influenzae isolates were serotyped with standard serological methods, and molecular typing was done with PCR. The capsular genotype of each isolate was characterized with PCR, partial sequencing, and Southern blot hybridization. Further strain typing was performed with pulsed field gel electrophoresis. RESULTS: We identified 2 children with severe invasive disease due to H. influenzae serotype a. Both H. influenzae serotype a isolates contained the identical pulsed-field gel electrophoresis pattern and capsular genotype. An IS1016-bexA partial deletion in the capsule gene locus similar to that found in H. influenzae serotype b was identified in both isolates by means of PCR and sequencing of the IS1016-bexA junction. CONCLUSIONS: We describe 2 cases of severe invasive disease due to H. influenzae serotype a with the putative virulence-enhancing IS1016-bexA partial deletion and duplication of the capsule locus. Our data support the hypothesis that this mutation may be associated with virulence in non-b capsular serotypes of H. influenzae. PMID- 16267726 TI - Acquired metallo-beta-lactamases: an increasing clinical threat. PMID- 16267725 TI - Dissemination of the metallo-beta-lactamase gene blaIMP-4 among gram-negative pathogens in a clinical setting in Australia. AB - BACKGROUND: The clinical utility of carbapenems is under threat because of the emergence of acquired metallo-beta-lactamase (MBL) genes. We describe the first outbreak in Australia of infection and/or colonization with gram-negative pathogens carrying the MBL gene blaIMP-4. METHODS: MBL-producing organisms were identified using susceptibility data in conjunction with MBL screening methods. PCR and sequence analysis were performed to characterize the resistance gene and identify the presence of integrons. DNA profiles were determined by ribotyping. Clinical and epidemiological data were prospectively collected from January-July 2004. RESULTS: A total of 19 isolates were recovered from 16 patients: Serratia marcescens (10 isolates), Klebsiella pneumoniae (4 isolates), Pseudomonas aeruginosa (3 isolates), Escherichia coli (1 isolate), and Enterobacter cloacae (1 isolate). Isolates were resistant to most beta-lactams except aztreonam, and variable resistance to carbapenems was observed (MIC range, 2 to >8 mg/L). PCR and sequence analysis identified the blaIMP-4 gene and a class 1 integrase (IntI1) in all isolates. Of the 16 patients, 12 (75%) had infection; 5 had septicemia, 5 had ventilator-associated pneumonia, 1 had a urinary tract infection, and 1 had a superficial central venous line infection. Six (38%) of the 16 patients died, and 5 of those 6 (31% of the group of 16) had clinical infection with an MBL-producing organism. All except 2 patients had spatio temporal epidemiological links in the intensive care unit. All K. pneumoniae isolates were of different ribogroups, whereas the S. marcescens and P. aeruginosa isolates were predominately of the same ribogroup. CONCLUSIONS: MBL producing gram-negative organisms have now emerged in Australia. The resistance gene, blaIMP-4, appears highly mobile; this outbreak involved 5 different gram negative genera from patients with close epidemiological links. PMID- 16267727 TI - Invasive Saccharomyces infection: a comprehensive review. AB - BACKGROUND: Saccharomyces cerevisiae (also known as "baker's yeast" or "brewer's yeast") is mostly considered to be an occasional digestive commensal. However, since the 1990s, there have been a growing number of reports about its implication as an etiologic agent of invasive infection. A particular feature of such infections is their association with a probiotic preparation of Saccharomyces boulardii (a subtype of S. cerevisiae) for treatment various diarrheal disorders. METHODS: We collected published case reports, through May 2005, of invasive Saccharomyces infection by use of a Medline query. Epidemiological and clinical charts and therapeutic strategies were analyzed. RESULTS: We found 92 cases of Saccharomyces invasive infection. Predisposing factors were similar to those of invasive candidiasis, with intravascular catheter and antibiotic therapy being the most frequent. Blood was the most frequent site of isolation (for 72 patients). S. boulardii accounted for 51.3% of fungemias and was exclusively isolated from blood. Compared with patients infected with S. cerevisiae, patients infected with S. boulardii were more frequently immunocompetent and had a better prognosis. Saccharomyces invasive infection was clinically indistinguishable from an invasive candidiasis. Overall, S. cerevisiae clinical isolates exhibited low susceptibility to amphotericin B and azole derivatives. However, global outcome was favorable in 62% of the cases. Treatment with intravenous amphotericin B and fluconazole, in combination with central vascular catheter removal, were effective therapeutic options. CONCLUSION: Saccharomyces organisms should now be added to the growing list of emerging fungal pathogens. Special caution should be taken regarding the use of S. boulardii probiotic preparations. PMID- 16267728 TI - Cervicofacial lymphadenitis in children caused by Mycobacterium haemophilum. AB - BACKGROUND: Nontuberculous mycobacterial (NTM) lymphadenitis in children is most often caused by Mycobacterium avium. In a prospective, multicenter trial of the optimal treatment, 23.7% of the NTM cervicofacial lymphadenitis cases in children were caused by Mycobacterium haemophilum. In this article, we describe the epidemiological and clinical features of M. haemophilum cervicofacial lymphadenitis. METHODS: The diagnosis of Mycobacterium avium or M. haemophilum infection was established by culture or polymerase chain reaction. Demographic characteristics and data regarding clinical presentation and possible environmental exposure were compared for patients infected with M. avium and those infected with M. haemophilum. RESULTS: Ninety-four (69.9%) of 135 infections were caused by M. avium, 32 (23.7%) by M. haemophilum, and 9 (6.4%) by other NTM species. The median age of the M. haemophilum-infected children was 72 months, compared with 41 months for the M. avium-infected children (P < .001), with an equal distribution for both sexes. Involvement of multiple lymph nodes was frequently observed among the M. haemophilum-infected patients (56% of patients). Extranodal localizations were only observed in M. haemophilum-infected patients. Children with M. haemophilum infection were more likely to have a non Dutch background (P = .001), and in most cases, they had a history of contact with swimming water (P = .03), whereas M. avium-infected patients were more likely to have a history of playing in sandpits (P = .01). In a multivariate analysis, only older age and a non-Dutch background were predisposing risk factors for M. haemophilum infection, compared with M. avium infection. CONCLUSION: Higher age, non-Dutch background, and involvement of multiple cervicofacial lymph nodes with extranodal localizations distinguished M. haemophilum infection from M. avium infection. PMID- 16267729 TI - Application of hepatitis B virus genotyping and phylogenetic analysis in intrafamilial transmission of hepatitis B virus. AB - BACKGROUND: Infection with hepatitis B virus (HBV) in early life frequently results in persistent infection, and clustering of the chronic infection within a family is common. However, the relative contribution of perinatal mother-to infant transmission or early horizontal transmission to the intrafamilial clustering of HBV infection remains unclear. Therefore, we used HBV genotyping and phylogenetic analysis to elucidate the modes of intrafamilial HBV transmission in Taiwan. METHODS: HBV genotypes and serological markers were determined for 103 individuals from 20 families with evidence of clustering HBV infection. RESULTS: Three patterns of intrafamilial clustering of HBV infection were identified. Among the 20 families, 8 included a hepatitis B surface antigen (HBsAg)-positive mother (pattern I), 7 included an HBsAg-positive father (pattern II), and in the remaining 5, both parents were positive for HBsAg (pattern III). The rates of HBsAg positivity for children of the 3 representative groups of families were 85.7%, 65.4%, and 87.5%, respectively (P = .16). The identical genotyping results between index parent and carrier children indicated that pattern I clustering was caused by maternal transmission, whereas pattern II clustering was caused by paternal transmission. In pattern III clustering, a concordant HBV genotype between carrier children and mother or father was found in 3 and 2 families, respectively. The modes of transmission were confirmed by phylogenetic analysis in 1 family of each pattern. CONCLUSIONS: In Taiwan, maternal and paternal transmissions are both important in the intrafamilial spread of HBV infection. PMID- 16267730 TI - Proton nuclear magnetic resonance-based metabonomics for rapid diagnosis of meningitis and ventriculitis. AB - BACKGROUND: Reduction of mortality associated with bacterial meningitis and postsurgical cerebral ventriculitis is dependent on early diagnosis and institution of appropriate therapy. Metabonomics rapidly defines metabolic profiles of biological fluids through the use of high-throughput analytical techniques combined with statistical pattern recognition tools. METHODS: Proton nuclear magnetic resonance (1H NMR)-based metabonomics was applied to (1) lumbar cerebrospinal fluid samples collected prospectively from a cohort of patients with bacterial, fungal, or viral meningitis and from control subjects without neurological disease and (2) ventricular cerebrospinal fluid samples from patients with ventriculitis associated with an external ventricular drain and from control subjects. 1H NMR spectra were analyzed by the unsupervised statistical method of principal components analysis. RESULTS: Metabonomic analysis clearly distinguished patients with bacterial or fungal meningitis (11 patients) from patients with viral meningitis (12) and control subjects (27) and clearly distinguished patients with postsurgical ventriculitis (5) from postsurgical control subjects (10). Metabolites of microbial and host origin that were responsible for class separation were determined. Metabonomic data also correlated with the onset and course of infection in a patient with 2 episodes of bacterial ventriculitis and with response to therapy in another patient with cryptococcal meningitis. CONCLUSIONS: Metabonomic analysis is rapid, requires minimal sample processing, and is not targeted to specific microbial pathogens, making the platform potentially suitable for use in the diagnostic laboratory. This pilot study indicates that metabonomic analysis of cerebrospinal fluid is feasible and a potentially more powerful diagnostic tool than conventional rapid laboratory indicators for distinguishing bacterial from viral meningitis and for monitoring therapy. This should have important implications for early management, reduced empirical use of antibiotics, and treatment duration. PMID- 16267732 TI - Tuberculosis, bronchiectasis, and infertility: what ailed George Orwell? AB - In the last and most productive years of his life, George Orwell struggled with pulmonary tuberculosis, dying at the dawn of the era of chemotherapy. His case history illustrates clinical aspects of tuberculosis with contemporary relevance: the role of poverty in its spread, the limited efficacy of monotherapy, the potential toxicity of treatment, and the prominence of cachexia as a terminal symptom. Orwell's ordeals with collapse therapy may have influenced the portrayal of the tortures of Winston Smith in the novel 1984. I discuss unifying diagnoses for Orwell's respiratory problems and apparent infertility, including tuberculous epididymitis, Young syndrome, immotile cilia syndrome, and cystic fibrosis. PMID- 16267731 TI - Clinical and economic outcomes in critically ill patients with nosocomial catheter-related bloodstream infections. AB - BACKGROUND: Central venous catheters are universally used during the treatment of critically ill patients. Their use, however, is associated with a substantial infection risk, potentially leading to increased mortality and costs. We evaluate clinical and economic outcomes associated with nosocomial central venous catheter related bloodstream infection (CR-BSI) in intensive care unit (ICU) patients. METHODS: A retrospective (1992-2002), pairwise-matched (ratio of case patients to control subjects, 1:2 or 1:1), risk-adjusted cohort study was performed at a 54 bed general ICU at a university hospital. ICU patients with microbiologically documented CR-BSI (n = 176) were matched with control subjects (n = 315) on the basis of disease severity, diagnostic category, and length of ICU stay (equivalent or longer) before the onset of CR-BSI in the index case patient. Clinical outcome was principally evaluated by in-hospital mortality. Economic outcome was evaluated on the basis of duration of mechanical ventilation, length of ICU and hospital stays, and total hospital costs, as derived from the patient's hospital invoices. RESULTS: The attributable mortality rate for CR-BSI was estimated to be 1.8% (95% confidence interval, -6.4% to 10.0%); in-hospital mortality rates for patients with CR-BSI and matched control subjects were 27.8% and 26.0%, respectively. CR-BSI was associated with significant excesses in duration of mechanical ventilation, duration of ICU and hospital stays, and a significant increase in total hospital cost. Linear regression analysis with adjustment for duration of hospitalization and clinical covariates, revealed that CR-BSI is independently associated with higher costs. CONCLUSIONS: In ICU patients, CR-BSI does not result in increased mortality. It is, however, associated with a significant economic burden, emphasizing the importance of continuous efforts in prevention. PMID- 16267733 TI - Chronic hepatitis C virus infection in older adults. AB - Most of the older adults with chronic hepatitis C virus infection acquired the disease earlier in life. These patients often present with complications of liver disease, mainly cirrhosis and hepatocellular carcinoma. The burden of chronic hepatitis C virus infection in elderly persons is expected to increase significantly in the United States during the next 2 decades. It seems important that, for elderly patients with chronic hepatitis C, the risk-benefit of combination antiviral therapy consisting of pegylated interferon and ribavirin should be assessed on an individual basis. Assessment should be performed in all cases before considering treatment, and it should include evaluation of the degree of liver fibrosis by means of liver biopsy or, possibly, by means of noninvasive methods. Novel antiviral drugs that may have fewer adverse effects, such as protease inhibitors, may serve as potential alternatives. It is recommended that elderly patients (up to the age of 75 years) be included in randomized trials of chronic hepatitis C virus infection treatment. PMID- 16267734 TI - Human health consequences of antimicrobial drug-resistant Salmonella and other foodborne pathogens. AB - There are several clinical and public health consequences associated with antimicrobial drug resistance in non-Typhi Salmonella species and other foodborne pathogens. If bacteria acquire resistance to clinically important antimicrobial drugs, early empirical treatment may fail, and there will be limitations in the choices of treatment after the establishment of microbial diagnosis. Drug resistant gastrointestinal pathogens preferentially cause illness in persons receiving antimicrobial drugs for any medical condition. Consequently, emerging resistance in foodborne pathogens may result in increased burdens of illness and outbreaks in settings where patients are treated with antimicrobial drugs. Finally, resistance may be associated with increased virulence, and several epidemiological studies have demonstrated that infections with drug-resistant non Typhi Salmonella serotypes and Campylobacter species are associated with excess mortality and morbidity. Mitigation of drug resistance in foodborne bacteria is likely to be of benefit for human health. PMID- 16267735 TI - Bacterial diarrhea in persons with HIV infection, United States, 1992-2002. AB - BACKGROUND: To describe trends in bacterial diarrhea among human immunodeficiency virus (HIV)-infected persons during 1992-2002, we examined data from a longitudinal record review study of persons with HIV infection who were receiving medical care in >100 medical facilities in 9 major United States cities. METHODS: An analysis was performed using data from 44,778 persons who were followed up for a mean of 2.6 years. We calculated incidence rates and rate ratios for bacterial diarrhea, by stage of HIV disease, and determined odds ratios (ORs) to compare bacterial diarrhea diagnosis in 2002 versus 1992. RESULTS: The mean annual incidence of bacterial diarrhea was 7.2 cases per 1000 person-years. The incidence of Clostridium difficile-associated diarrhea, the most common bacterial cause of diarrhea, was 4.1 cases per 1000 person-years. Compared with persons without AIDS, persons with AIDS were more likely to have bacterial diarrhea (incidence rate ratio, 1.3-9.9, varying by clinical versus immunologic AIDS and type of bacterial diarrhea). Between 1992 and 2002, the overall rate of bacterial diarrhea in persons with clinical AIDS decreased (OR, 0.4; 95% confidence interval, 0.2-0.6). During the same period, bacterial diarrhea rates among other persons in the analysis did not significantly change. CONCLUSIONS: C. difficile is the most common recognized cause of bacterial diarrhea among persons infected with HIV. The risk for bacterial diarrhea increases with increased severity of HIV disease. Health care professionals should be aware that patients with AIDS are at increased risk for bacterial diarrhea, and they should reinforce recommendations for decreasing the chances of acquiring bacterial diarrhea. PMID- 16267736 TI - Bacterial diarrhea in HIV-infected patients: why Clostridium difficile, and why now? PMID- 16267737 TI - Increased prevalence of severe malaria in HIV-infected adults in South Africa. AB - BACKGROUND: Conflicting reports exist regarding the impact of human immunodeficiency virus (HIV) infection on the risk of severe malaria. We aimed to assess the effect of HIV infection status, advancing immunosuppression, and antimalarial immunity on the severity of malaria. METHODS: A prospective cohort study was conducted. Consecutive hospitalized adult patients with falciparum malaria were tested for HIV antibodies and to determine CD4+ T cell count. Immunity to malaria was assessed by obtaining a history of childhood residence in an area where malaria is endemic. Patients were assessed for features of severe malaria. RESULTS: Three hundred thirty-six patients were enrolled in the study, of whom 32 (10%) had severe malaria. The prevalence of HIV infection was 33%, and 111 patients (33%) were nonimmune to malaria. HIV-infected patients complained more frequently about respiratory and abdominal symptoms and less frequently about rigors and headache. Risk factors for severe malaria determined by multivariate analysis included being nonimmune to malaria, having a positive HIV serostatus, having an elevated parasite count, and having an increased white blood cell count. Risk of severe malaria was increased in HIV-infected patients with a CD4+ T cell count of < 200 x 10(6) cells/L (P < or = .001). Nonimmune HIV infected patients were significantly more likely to have severe malaria (13 [36%] of 36 patients) than were nonimmune non-HIV-infected patients (9 [12%] of 75 patients; odds ratio, 4.15 [95% confidence interval, 1.57-10.97]; P = .003). HIV serostatus did not affect risk of severe malaria in the group from an area with endemicity (5 [7%] of 74 HIV-infected patients had severe malaria, and 5 [3%] of 151 non-HIV-infected patients had malaria; P = .248). CONCLUSIONS: HIV-infected nonimmune adults are at increased risk of severe malaria. This risk is associated with a low CD4+ T cell count. This interaction is of great public health importance. PMID- 16267738 TI - The clinical pharmacokinetics of rifampin and ethambutol in HIV-infected persons with tuberculosis. AB - BACKGROUND: The pharmacokinetics of rifampin and ethambutol in HIV-infected patients with tuberculosis (TB) are incompletely characterized. We examined the pharmacokinetics of rifampin and ethambutol in a cohort of patients with HIV related TB who were treated in the United States. METHODS: Serum drug concentrations were determined 2, 6, and 10 h after dosing in 36 HIV-infected patients with TB who were taking rifampin and in 49 who were taking ethambutol. Observed serum concentrations were compared with published normal ranges and published data. RESULTS: With daily dosing of rifampin (600 mg), 26 (77%) of 34 patients (95% confidence interval [CI], 59%-89%]) had a low maximum concentration of rifampin (<8 microg/mL), and 12 (35%; 95% CI, 20%-54%) had a very low maximum concentration (<4 microg/mL). With intermittent rifampin dosing (600 mg), 13 (68%) of 19 patients (95% CI, 44%-85%) had a low maximum concentration of rifampin, and 5 (26%; 95% CI, 11%-50%) had a very low maximum concentration. With daily ethambutol dosing (20 mg/kg), 33 (69%) of 48 patients (95% CI, 55%-81%) had a low maximum concentration of ethambutol (<2 microg/mL), and 18 (38%; 95% CI, 24%-53%) had a very low maximum concentration (<1 microg/mL). With intermittent ethambutol dosing (50 mg/kg twice weekly or 30 mg/kg thrice weekly), 13 (72%) of 18 patients (95% CI, 47%-88%) had a low maximum concentration of ethambutol (<4 microg/mL), and 5 (28%; 95% CI, 12%-54%]) had a very low maximum concentration (<2 microg/mL). CONCLUSIONS: In HIV-infected patients with TB who are receiving rifampin and ethambutol, low maximum concentrations of rifampin and ethambutol were common. For patients with HIV-related TB, therapeutic monitoring of rifampin and ethambutol levels may help clinicians achieve target serum concentrations. PMID- 16267739 TI - Prediction of neuropsychiatric adverse events associated with long-term efavirenz therapy, using plasma drug level monitoring. AB - BACKGROUND: Data on long-term central nervous system (CNS) toxicity associated with efavirenz therapy are scarce, and risk factors remain largely unknown. We aimed to determine whether monitoring the plasma concentration of efavirenz could predict neuropsychiatric adverse events associated with long-term therapy with efavirenz. METHODS: We performed a longitudinal study involving 17 consecutive human immunodeficiency virus (HIV)-infected subjects with virological suppression after at least 6 months of antiretroviral therapy with an efavirenz-containing regimen. Efavirenz plasma concentrations were measured at study entry and at different time points through an 18-month study period. RESULTS: Median duration of efavirenz therapy before study entry was 18 months (range, 6-27 months). Ten (58.8%) of the patients experienced CNS-related adverse effects, ranging from insomnia and abnormal dreams to depression with suicidal ideation. In 4 (23.5%) of the cases, CNS toxicity led to efavirenz discontinuation. Mean (+/- standard deviation) plasma levels were higher for patients experiencing neuropsychiatric symptoms (5.10 +/- 2.15 microg/mL vs. 2.79 +/- 1.31 microg/mL; P = .024). A plasma level of 2.74 microg/mL had a sensitivity of 90.9% and specificity of 72% to predict CNS toxicity (area under the curve, 0.839; 95% confidence interval, 0.73-0.95; P < .0001). Patients having efavirenz plasma concentrations > 2.74 microg/mL at any time point of the study were 5.68 times more likely to experiencing CNS toxicity than were other patients (95% confidence interval, 1.97 16.37). CONCLUSIONS: In patients with HIV infection receiving long-term therapy with efavirenz-containing antiretroviral regimens, CNS toxicity is related to efavirenz plasma levels. Patients achieving higher plasma levels are at increased risk of experiencing neuropsychiatric adverse events. PMID- 16267740 TI - Does severity of HIV disease in HIV-infected mothers affect mortality and morbidity among their uninfected infants? AB - BACKGROUND: Rates of perinatal human immunodeficiency virus (HIV) transmission are higher among HIV-infected mothers with more advanced disease, but effects of maternal disease on HIV-uninfected offspring are unclear. We investigated the hypothesis that the severity of HIV disease and immune dysfunction among mothers is associated with increased morbidity and mortality among their uninfected infants. METHODS: In a birth cohort of 620 HIV-uninfected infants born to HIV infected mothers in Lusaka, Zambia, we investigated associations between markers of more advanced maternal HIV disease and child mortality, hospital admissions, and infant weight through 4 months of age. RESULTS: Mortality in the cohort of uninfected infants was 4.6% (95% confidence interval [CI], 2.8-6.3) through 4 months of age. Infants of mothers with CD4+ T cell counts of <350 cells/microL were more likely to die (hazard ratio [HR], 2.87; 95% CI, 1.03-8.03) and were more likely to be hospitalized (HR, 2.28; 95% CI, 1.17-4.45), after adjusting for other factors, including maternal death and low birth weight. The most common cause of infant death and hospitalization was pneumonia and/or sepsis. A maternal viral load of >100,000 copies/mL was associated with significantly lower child weight through 4 months of age. CONCLUSION: Children born to HIV-infected mothers with advanced disease who escaped perinatal or early breastfeeding-related HIV infection are nonetheless at high risk of mortality and morbidity during the first few months of life. HIV-related immunosuppression appears to have adverse consequences for the health of infants, in addition to risks of vertical transmission. PMID- 16267741 TI - CD4+ cell count, viral load, and highly active antiretroviral therapy use are independent predictors of body composition alterations in HIV-infected adults: a longitudinal study. AB - BACKGROUND: To understand the concurrent effects of human immunodeficiency virus (HIV) infection, the immune system, and antiretroviral therapy on body composition alterations, we examined annualized composition changes in HIV infected adults who were receiving stable antiretroviral therapy. METHODS: With use of data from the Nutrition For Healthy Living Study, we performed multivariate analyses using longitudinal models to evaluate the relationship of CD4+ cell count, viral load, and highly active antiretroviral therapy (HAART) or antiretroviral therapy (ART) with changes in trunk and extremity composition for 110 men and 42 women who provided data relating to 194 study intervals (i.e., intervals of time between 2 assessment visits). Of these intervals, 165 involved HAART use (89.7% involved protease inhibitor-based regimens), and 29 did not involve HAART use. Patients receiving HAART or ART (who had continuous use during the interval) were compared with HAART- or ART-naive subjects. RESULTS: The median length of intervals between visits was 12.9 months (interquartile range, 12.1-17.6 months). In models adjusted for HAART or ART use, baseline CD4+ cell count was positively associated with increased trunk fat (mean increase per year, 2.3% per 100 cells/mm3; 95% confidence interval [CI], 0.7%-3.9%]) and, in men, with increased extremity fat (mean increase per year, 1.8% per 100 cells/mm3; 95% CI, 0.6%-3.0%). Increase in CD4+ cell count predicted increased extremity lean mass (mean increase per year, 0.6% per 100 cells/mm3; 95% CI, 0.05%-1.1%). Higher baseline viral load predicted fat loss (trunk fat loss per year, -5.0% per log10 copies/mL; 95% CI, -9.4% to -0.7%; extremity fat loss per year, -3.4% per log10 copies/mL; 95% CI, -6.1% to -0.6%), as did zidovudine use (trunk fat loss per year, -10.8%; 95% CI, -20.4% to -1.4%; extremity fat loss per year, -4.9%; 95% CI, -9.8% to -0.01%). HAART use independently predicted decreased bone mineral content (extremity bone mineral content loss per year, -1.6%; 95% CI, -3.1% to 0.08%) but did not predict changes in fat or lean mass. Receipt of protease inhibitor-based HAART predicted a -1.9% decrease in extremity bone mineral content per year (95% CI, -3.6% to -0.2%), and zidovudine use predicted a -2.6% decrease in trunk bone mineral content per year (95% CI, -4.4% to -0.8%). CONCLUSIONS: Baseline viral load, CD4+ cell count, and change in CD4+ cell count predicted alterations in trunk fat, extremity fat, and lean mass. HAART use and zidovudine use were associated with bone loss, and zidovudine use was associated with fat loss, but HAART use was not associated with fat mass changes. PMID- 16267742 TI - Transient viremia in HIV-infected patients and use of plasma preparation tubes. AB - Using plasma preparation tubes for the collection and storage of plasma resulted in factitious, low-level human immunodeficiency virus type 1 (HIV-1) viremia among patients receiving highly active antiretroviral therapy who incurred unnecessary additional clinic visits, laboratory testing, and medication changes. We caution clinicians against the routine use of plasma preparation tubes for collection of blood samples for HIV-1 level 1 quantification. PMID- 16267743 TI - Prolonged incubation and extensive subculturing do not increase recovery of clinically significant microorganisms from standard automated blood cultures. AB - An extensive blood culture protocol, including prolonged incubation of cultures, for 215 patients believed to have had endocarditis yielded only 3 clinically relevant results. Twenty-four Haemophilus, Actinobacillus, Cardiobacterium, Eikenella, and Kingella (i.e., HACEK) organisms were recovered from standard 5 day blood cultures during the same time period. Specialized methods and not extended incubation times are recommended for recovery of fastidious agents of septicemia. PMID- 16267744 TI - Emerging data indicating that extended incubation of blood cultures has little clinical value. PMID- 16267745 TI - Efficacy of antiretroviral therapy in resource-poor settings: are outcomes comparable to those in the developed world? PMID- 16267747 TI - Listeria monocytogenes meningitis following imatinib mesylate-induced monocytopenia in a patient with chronic myeloid leukemia. PMID- 16267748 TI - Infliximab does not affect dendritic cells' mediated lymphoproliferative response to Histoplasma capsulatum. PMID- 16267750 TI - Misattribution of central nervous system dysfunction to artesunate. PMID- 16267752 TI - Medical treatment of fish bone-related liver abscess. PMID- 16267753 TI - Fluconazole prophylaxis for critically ill patients at high risk for Candida infection. PMID- 16267754 TI - Nature usually favors females. PMID- 16267755 TI - Less than the sum of its parts: failure of a tenofovir-abacavir-Lamivudine triple nucleoside regimen. PMID- 16267756 TI - Intermittent preventive treatment for malaria in infants: moving forward, cautiously. PMID- 16267757 TI - A significant sex--but not elective cesarean section--effect on mother-to-child transmission of hepatitis C virus infection. AB - BACKGROUND: Risk factors for mother-to-child transmission of hepatitis C virus (HCV) are poorly quantified. METHODS: We conducted a European multicenter prospective study of HCV-infected pregnant women and their infants. Children with > or =2 positive HCV RNA polymerase chain reaction test results and/or anti-HCV antibodies after 18 months of age were considered to be infected. RESULTS: The overall HCV vertical transmission rate was 6.2% (95% confidence interval [CI], 5.0%-7.5%; 91/1479). Girls were twice as likely to be infected as boys (adjusted odds ratio [OR], 2.07 [95% CI, 1.23-3.48]; P=.006). There was no protective effect of elective cesarean section (CS) delivery on HCV vertical transmission (adjusted OR, 1.46 [95% CI, 0.86-2.48]; P=.16). HCV/human immunodeficiency virus coinfected women more frequently transmitted HCV than did women with HCV infection only, although the difference was not statistically significant (adjusted OR, 1.82 [95% CI, 0.94-3.52]; P=.08). Maternal history of injection drug use, prematurity, and breast-feeding were not significantly associated with transmission. Transmission occurred more frequently from viremic women, but it also occurred from a few nonviremic women. CONCLUSIONS: Our results strongly suggest that women should neither be offered an elective CS nor be discouraged from breast-feeding on the basis of HCV infection alone. The sex association is an intriguing finding that probably reflects biological differences in susceptibility or response to infection. PMID- 16267758 TI - Risk factors for perinatal transmission of hepatitis C virus (HCV) and the natural history of HCV infection acquired in infancy. AB - BACKGROUND: The goal of the present study was to assess risk factors for perinatal hepatitis C virus (HCV) transmission and the natural history of infection among HCV-infected infants. METHODS: In a cohort study, 244 infants born to HCV-positive mothers were followed from birth until age > or =12 months. Maternal serum was collected at enrollment and delivery; infant serum was collected at birth and at 8 well-child visits. Testing included detection of antibody to HCV, detection of HCV RNA (qualitative and quantitative), and genotyping. HCV-infected infants were followed annually until age 5 years. RESULTS: Overall, 9 of 190 (4.7% [95% confidence interval (CI), 2.3%-9.1%]) infants born to mothers who were HCV RNA positive at delivery became infected, compared with 0 of 54 infants born to HCV RNA-negative mothers (P=.10). Among HCV RNA-positive mothers, the rate of transmission was 3.8% (95% CI, 1.7%-8.1%) from the 182 who were human immunodeficiency virus (HIV) negative, compared with 25.0% (95% CI, 4.5%-64.4%) from the 8 who were HIV positive (P<.05). Three infected infants resolved their infection (i.e., became HCV RNA negative). In multivariate analysis restricted to HCV RNA-positive mothers, membrane rupture > or =6 h (odds ratio [OR], 9.3 [95% CI, 1.5-179.7]) and internal fetal monitoring (OR, 6.7 [95% CI, 1.1-35.9]) were associated with transmission of HCV to infants. CONCLUSION: If duration of membrane rupture and internal fetal monitoring are confirmed to be associated with transmission, interventions may be possible to decrease the risk of transmission. PMID- 16267759 TI - Reducing the global burden of congenital rubella syndrome: report of the World Health Organization Steering Committee On Research Related To Measles and Rubella Vaccines and Vaccination, June 2004. AB - Rubella and congenital rubella syndrome (CRS) continue to be important health problems in many countries. In June 2004, the World Health Organization Steering Committee on Research Related to Measles and Rubella Vaccines and Vaccination met to evaluate data from research and operational activities and to identify critical scientific issues and gaps in knowledge that need to be addressed to improve the global control of rubella and CRS. Information about surveillance for rubella, natural and vaccine-induced immunity to rubella, laboratory diagnosis, the molecular epidemiological profile of rubella virus, and mathematical modeling to assess the burden of CRS and the impact of rubella vaccination was reviewed. This report summarizes the presentations and recommendations for future research. PMID- 16267761 TI - The granzyme B inhibitor SERPINB9 (protease inhibitor 9) circulates in blood and increases on primary cytomegalovirus infection after renal transplantation. AB - SERPINB9 is the only known human intracellular inhibitor of granzyme B (GrB), the effector molecule in immunity against cytomegalovirus (CMV) and in renal allograft rejection. Therefore, using specific enzyme-linked immunosorbent assays, we addressed the presence of circulating SERPINB9 during primary CMV infection, subclinical rejection, acute rejection, and uncomplicated posttransplantation course. Soluble (s) SERPINB9 circulates in blood and increases on primary CMV infection. This increase was significantly higher in symptomatic than in asymptomatic patients. In contrast, sSERPINB9 levels did not change in response to subclinical or acute rejection. We demonstrated the presence of circulating sSERPINB9/sGrB complexes, which suggests that SERPINB9 has extracellular functions as well. PMID- 16267762 TI - Simultaneous generation of cytomegalovirus-specific CD8+ and CD4+ T lymphocytes by use of dendritic cells comodified with pp65 mRNA and pp65 protein. AB - Cytomegalovirus (CMV) disease remains a severe complication in patients who have undergone transplantation. Viremia can be prevented and treated by the adoptive transfer of donor-derived CMV-directed T cells. To ensure long-term protection against CMV disease, it is important to transfer CMV antigen-specific T cells that represent both the CD8+ and the CD4+ subsets. In the present study, we used as stimulators dendritic cells (DCs) that were electroporated with in vitro transcribed 5'-capped polyadenylated messenger RNA (mRNA) that encoded the CMV pp65 protein (i.e., pp65 mRNA). These DCs could efficiently activate CMV-directed CD8+ T cells, as assayed by tetramer staining, interferon- gamma production, and cytolytic activity. We also used DCs that were pulsed with a recombinant pp65 protein to activate CMV-directed CD4+ T cells. When DCs were comodified with pp65 mRNA and pp65 protein, large numbers of CMV-directed CD8+ and CD4+ T cells were generated simultaneously. The approach outlined in the present study can be adapted for a clinical protocol that circumvents potential virus-related biohazards and is available to all patients independently of their human leukocyte antigen haplotype. PMID- 16267760 TI - Clinical and molecular epidemiological features of coronavirus HKU1-associated community-acquired pneumonia. AB - BACKGROUND: Recently, we described the discovery of a novel group 2 coronavirus, coronavirus HKU1 (CoV-HKU1), from a patient with pneumonia. However, the clinical and molecular epidemiological features of CoV-HKU1-associated pneumonia are unknown. METHODS: Prospectively collected (during a 12-month period) nasopharyngeal aspirates (NPAs) from patients with community-acquired pneumonia from 4 hospitals were subjected to reverse-transcription polymerase chain reaction, for detection of CoV-HKU1. The epidemiological, clinical, and laboratory characteristics of patients with CoV-HKU1-associated pneumonia were analyzed. The pol, spike (S), and nucleocapsid (N) genes were also sequenced. RESULTS: NPAs from 10 (2.4%) of 418 patients with community-acquired pneumonia were found to be positive for CoV-HKU1. All 10 cases occurred in spring and winter. Nine of these patients were adults, and 4 had underlying diseases of the respiratory tract. In the 6 patients from whom serum samples were available, all had a 4-fold change in immunoglobulin (Ig) G titer and/or presence of IgM against CoV-HKU1. The 2 patients who died had significantly lower hemoglobin levels, monocyte counts, albumin levels, and oxygen saturation levels on admission and had more-extensive involvement visible on chest radiographs. Sequence analysis of the pol, S, and N genes revealed 2 genotypes of CoV-HKU1. CONCLUSIONS: CoV-HKU1 accounts for 2.4% of community-acquired pneumonia, with 2 genotypes in the study population. Without performance of diagnostic tests, the illness was clinically indistinguishable from other community-acquired pneumonia illnesses. PMID- 16267763 TI - Early virologic nonresponse to tenofovir, abacavir, and lamivudine in HIV infected antiretroviral-naive subjects. AB - BACKGROUND: Antiretroviral combinations that reduce the number of pills and dosing frequency have the potential to simplify therapy. We compared 2 regimens dosed as 2 pills once daily. METHODS: This was a randomized, open-label, multicenter study of tenofovir disoproxil fumarate versus efavirenz, both administered once daily with the abacavir/lamivudine fixed-dose combination in treatment-naive human immunodeficiency virus type 1 (HIV-1)-infected subjects. After reports of early nonresponse, an unplanned interim analysis was performed. Virologic nonresponse was defined as (1) a <2.0-log(10) copies/mL decrease in HIV 1 RNA level by week 8, (2) an HIV-1 RNA rebound of > or =1.0 log(10) copies/mL above the nadir, or (3) for subjects with 2 consecutive HIV-1 RNA measurements <50 copies/mL, a subsequent increase to >400 copies/mL on 2 consecutive occasions. RESULTS: We randomized 340 subjects. Median baseline HIV-1 RNA level and CD4+ cell count were 4.7 log(10) copies/mL and 251 cells/mm3, respectively; 194 subjects with HIV-1 RNA data from > or =8 weeks were included in the interim analysis. Virologic nonresponse occurred in 50 (49%) of 102 subjects in the tenofovir disoproxil fumarate arm, compared with 5 (5%) of 92 of subjects in the efavirenz arm (P<.001). Within 12 weeks, viral genotypes for nonresponders in the tenofovir disoproxil fumarate arm showed M184V or I/M/V mixtures in 40 (98%) of 41 subjects and K65R and M184V or mixtures in 22 (54%) of 41 subjects. The protocol was immediately amended to modify the tenofovir disoproxil fumarate arm. The efavirenz arm continued unchanged; after 48 weeks, 120 (71%) of 169 subjects achieved HIV-1 RNA levels <50 copies/mL. CONCLUSION: The tenofovir disoproxil fumarate/abacavir/lamivudine regimen resulted in an unexpected and unacceptably high rate of nonresponse and incidence of K65R and M184V/I. This 3-drug regimen should not be used. PMID- 16267765 TI - Histological findings and clinical characteristics associated with hepatic steatosis in patients coinfected with HIV and hepatitis C virus. AB - BACKGROUND: Hepatic steatosis, a common histological finding in hepatitis C virus (HCV)-infected patients, is associated with severity of fibrosis. The prevalence and significance of steatosis in patients coinfected with human immunodeficiency virus (HIV) and HCV are not well characterized. METHODS: To determine the prevalence and severity of steatosis, a single pathologist evaluated liver-biopsy samples from 106 patients coinfected with HIV and HCV but without hepatitis B infection (negative results for hepatitis B surface antigen) for findings associated with steatosis or steatohepatitis and viral hepatitis. Medical records were reviewed retrospectively to elucidate risk factors for steatosis. RESULTS: Steatosis was present in 56% of biopsy samples, with moderate to severe grades in 9%. Severity of steatosis was associated with fibrosis (odds ratio [OR], 1.84 [95% confidence interval (CI), 1.06-3.20]; P=.03) but not with necroinflammation. In multivariate analysis, the severity of steatosis was associated with lower levels of high-density lipoprotein cholesterol (OR, 0.71 per 10-mg/dL increase [95% CI, 0.52-0.95]; P=.02), higher body-mass index (OR, 1.30 per kg/m2 increase [95% CI, 1.13-1.49]; P<.001), and the presence of lipodystrophy (OR, 3.82 [95% CI, 1.13-12.88]; P=.03). There was a trend toward an association between the severity of steatosis and fibrosis in multivariate analysis (OR, 1.69 [95% CI, 0.91-3.16]; P=.10). CONCLUSIONS: In patients coinfected with HIV and HCV, hepatic steatosis is common and associated with more-advanced fibrosis. Lower levels of high-density lipoprotein cholesterol, higher body-mass index, and lipodystrophy are potentially modifiable risk factors associated with the severity of steatosis. PMID- 16267764 TI - Pharmacogenetics of long-term responses to antiretroviral regimens containing Efavirenz and/or Nelfinavir: an Adult Aids Clinical Trials Group Study. AB - BACKGROUND: Efavirenz and nelfinavir are metabolized by cytochrome P-450 (CYP) 2B6 and CYP2C19, respectively, with some involvement by CYP3A. Nelfinavir is a substrate for P-glycoprotein, which is encoded by MDR1. The present study examined associations between genetic variants and long-term responses to treatment. METHODS: Adult AIDS Clinical Trials Group study 384 randomized antiretroviral-naive subjects to receive efavirenz and/or nelfinavir plus 2 nucleoside analogues, with follow-up lasting up to 3 years. Population pharmacokinetics were estimated from a nonlinear mixed-effects model. Polymorphisms in CYP2B6, CYP2C19, CYP3A4, CYP3A5, and MDR1 were characterized. RESULTS: The 504 participants in the genetic study included 340 efavirenz recipients and 348 nelfinavir recipients (184 of the 504 participants received both efavirenz and nelfinavir). Of the participants, 49% were white, 31% were black, and 19% were Hispanic. Plasma exposure to efavirenz and nelfinavir in each population was significantly associated with the polymorphisms CYP2B6 516G-->T and CYP2C19 681G-->A, respectively. Among efavirenz recipients, the MDR1 position 3435 TT genotype was associated with decreased likelihood of virologic failure and decreased emergence of efavirenz-resistant virus but not with plasma efavirenz exposure. Among nelfinavir recipients, a trend toward decreased virologic failure was associated with the polymorphism CYP2C19 681G-->A. CONCLUSIONS: Genetic variants predict plasma exposure to efavirenz and nelfinavir, and they may predict virologic failure and/or emergence of drug resistant virus. These associations with treatment responses must be validated in other studies. PMID- 16267766 TI - Influence of HIV infection on changes in circulating leukocyte counts during measles in Zambian children. AB - Measles remains an important problem in Africa, where human immunodeficiency virus type 1 (HIV-1) infection is prevalent. To identify the consequences of coinfection, Zambian children hospitalized with measles were studied at entry, discharge, and 1 month after discharge. All children had low lymphocyte and eosinophil counts at entry and high leukocyte and monocyte counts during recovery. The death of cultured lymphocytes was more prolonged for HIV-positive children. CD38 and Fas were increased on CD4+ and CD8+ lymphocytes, and CD28 was decreased on CD8+ lymphocytes in all children. Abnormalities in CD4+ and CD8+ lymphocyte percentages and CD28 expression associated with HIV infection were preserved during measles. Therefore, the patterns of changes in leukocyte counts were similar, with little evidence that measles exacerbated HIV-associated lymphocyte abnormalities. PMID- 16267767 TI - Schistosomiasis and HIV-1 infection in rural Zimbabwe: effect of treatment of schistosomiasis on CD4 cell count and plasma HIV-1 RNA load. AB - To determine whether treatment of schistosomiasis has an effect on the course of human immunodeficiency virus type 1 (HIV-1) infection, individuals with schistosomiasis and with or without HIV-1 infection were randomized to receive praziquantel treatment at inclusion or after a delay of 3 months; 287 participants were included in the study, and 227 (79%) were followed up. Among the 130 participants who were coinfected, those who received early treatment (n=64) had a significantly lower increase in plasma HIV-1 RNA load than did those who received delayed treatment (n=66) (P<.05); this difference was associated with no change in plasma HIV-1 RNA load in the early intervention group (P=.99) and an increase in plasma HIV-1 RNA load in the delayed intervention group (P<.01). Among the 227 participants who were followed up, those who received early treatment (n=105) had an increase in CD4 cell count, whereas those who received delayed treatment (n=122) did not (P<.05); this effect did not differ between participants when stratified by HIV-1 infection status (P=.17). The present study suggests that treatment of schistosomiasis can reduce the rate of viral replication and increase CD4 cell count in the coinfected host. PMID- 16267768 TI - Parasitological rebound effect and emergence of pyrimethamine resistance in Plasmodium falciparum after single-dose sulfadoxine-pyrimethamine. AB - Intermittent preventive treatment for malaria in infants (IPTi) is a promising malaria control strategy. However, mass preventive treatment for malaria inherently bears the risk of increasing drug resistance. Here, the effect of single-dose sulfadoxine-pyrimethamine (S-P) versus placebo on Plasmodium falciparum infection rates was assessed in 63 selected infants who were aparasitemic at enrollment. An increase in the proportion of infants with isolates exhibiting drug resistance-associated mutations was detected 3 weeks after drug application in the treatment group. S-P, in the setting of IPTi, appears to cause a parasitological rebound effect in which there is selection of drug-resistant parasites for a short period after drug clearance. PMID- 16267770 TI - Influence of carriage of hemoglobin AS and the Fc gamma receptor IIa-R131 allele on levels of immunoglobulin G2 antibodies to Plasmodium falciparum merozoite antigens in Gabonese children. AB - BACKGROUND: To extend our previous findings showing an imbalanced distribution of immunoglobulin G2 (IgG2) antibodies to Plasmodium falciparum merozoite surface protein 2 (MSP2) and a higher frequency of infection with multiple P. falciparum strains in Gabonese children with sickle cell trait (hemoglobin AS), human Fc gamma receptor (Fc gamma R) IIa (CD32) polymorphism and the rate of in vitro invasion of red blood cells (RBCs) from subjects with either hemoglobin AA or AS by multiple P. falciparum strains were investigated. METHODS: Fc gamma RIIa mutation at amino acid position 131 (arginine or histidine) was detected by polymerase chain reaction, and in vitro cultures for parasites were used to assess the invasion rate. RESULTS: Fc gamma RIIa polymorphism is normally distributed in this population, with no preferential carriage by children with hemoglobin AS. Lower levels of IgG2 subclass antibodies to MSP2 peptides were independently associated with the Fc gamma RIIa-R131 allele and with carriage of hemoglobin AS. Our data suggest that IgG3 antibody responses to MSP2 epitopes could be exacerbated by lower IgG2 levels in children with hemoglobin AS. CONCLUSIONS: The higher rate of invasion of RBCs in the presence of multiple strains may indicate that several invasion pathways are solicited simultaneously, and the longer persistence of ring forms in RBCs from the subjects with hemoglobin AS might reflect a slower multiplication phase, leading to a longer circulation and enhanced phagocytosis of these nonpathogenic parasite forms. This may contribute to the protection against P. falciparum malaria observed in children with hemoglobin AS. PMID- 16267771 TI - The predictive validity of naturally acquired delayed-type hypersensitivity to leishmanin in resistance to Leishmania major-associated cutaneous leishmaniasis. AB - To accurately quantify the different outcomes of Leishmania major infection and to evaluate the fraction of zoonotic cutaneous leishmaniasis (ZCL) cases prevented by naturally acquired leishmanin skin test (LST) reactivity, a cohort of 470 children was followed up in 2 endemic foci, Remada and Dhiba, in southern Tunisia. During May 1997, before the ZCL emergence season, LST was performed, and results were reassessed 12 months later. Active case detection during the ZCL emergence season showed a high incidence of ZCL: 57.0% in Remada and 13.7% in Dhiba. The preventive fraction of ZCL conferred by LST reactivity increased proportionally with the reaction size before the emergence season, revealing a dose-response effect of approximately 70%. In addition, asymptomatic L. major infection appeared to be a significant form of natural immunization, particularly in the context of relatively low transmission. These findings may help in the design and evaluation of vaccines. PMID- 16267769 TI - Lung injury in uncomplicated and severe falciparum malaria: a longitudinal study in papua, Indonesia. AB - BACKGROUND: In patients with severe malaria, acute respiratory distress syndrome usually develops after the start of drug treatment and is a major cause of death. Its pathogenesis is not well understood. METHODS: Respiratory symptom, spirometry, and gas transfer analyses were performed longitudinally in adults in Papua, Indonesia, with uncomplicated (n=50) and severe (n=30) falciparum malaria; normal values were derived from 109 control subjects. Gas transfer was partitioned into its alveolar-capillary membrane (D(M)) and pulmonary vascular (Vc) components, to characterize the site of impaired gas transfer. RESULTS: Cough was frequent in both patients with uncomplicated malaria (50%) and those with severe malaria (30%) and resolved by day 14. Reduced midexpiratory flow indicated obstruction of the small airways. Gas transfer was significantly impaired in patients with severe malaria. D(M) was reduced in patients with severe malaria but not in those with uncomplicated malaria and only returned to normal levels after 2 weeks. In patients with uncomplicated malaria, Vc was reduced at presentation but improved thereafter. In patients with severe malaria, Vc decreased with treatment and was lowest at day 7. CONCLUSIONS: Our results suggest that pulmonary vascular occlusion occurs in both patients with uncomplicated malaria and those with severe malaria, likely from sequestration of both red blood cells (RBCs) and white blood cells. There was also impaired alveolar-capillary membrane function in patients with severe malaria but not in those with uncomplicated malaria. Persistent impairment long after clearance of parasitized RBCs suggests prolonged posttreatment inflammatory alveolar-capillary injury. PMID- 16267772 TI - Postvaccine genetic structure of Streptococcus pneumoniae serotype 19A from children in the United States. AB - BACKGROUND: The introduction of the 7-valent conjugate pneumococcal vaccine (PCV7) in children may result in serotype replacement. We estimated the rate of increase of invasive pneumococcal disease (IPD) caused by serotype 19A in children <5 years old and determined the genetic composition of these isolates. METHODS: Cases of IPD between July 1999 and June 2004 were identified through the Active Bacterial Core Surveillance. Serotype 19A isolates obtained from children <5 years old between January 2003 and June 2004 were characterized by serotyping, antibiotic susceptibility testing, and pulsed-field gel electrophoresis (PFGE). Select isolates representing homologous PFGE clusters were subjected to multilocus sequence typing, and eBURST was used to delineate clonal groups. RESULTS: Between July 1999 and June 2004, the overall rate of IPD decreased from 23.3 to 13.1 cases/100,000 population (P<.00001). In children <5 years old, the rate decreased from 88.7 to 22.4 cases/100,000 population (P<.00001), whereas the rate in persons > or =5 years old decreased from 18.4 to 12.4 cases/100,000 population (P<.0001). The rate of serotype 19A IPD in children <5 years old increased significantly from 2.6 cases/100,000 population in 1999-2000 to 6.5 cases/100,000 population in 2003-2004; this was accompanied by significant increases in penicillin nonsusceptibility (P=.008) and multidrug resistance (P=.002) among serotype 19A isolates. As was observed during the pre-PCV7 era, clonal complex (CC) 199 predominated within serotype 19A, representing approximately 70% of invasive serotype 19A isolates from children <5 years old during 2003-2004. New serotype 19A genotypes were observed during 2003-2004, including 6 CCs that were not found among pneumococcal serotype 19A isolates during surveillance in 1999. CONCLUSION: Serotype 19A is, at present, the most important cause of IPD by replacement serotypes, and it is increasingly drug resistant. CC199 is the predominant CC among type 19A serotypes in children <5 years old. Our data suggest that some of the increase in rates of infection with serotype 19A may be due to serotype switching within certain vaccine type strains. PMID- 16267773 TI - In vivo binding of complement regulator factor H by Streptococcus pneumoniae. AB - Pneumococcal surface protein C (PspC) binds to the complement regulatory protein factor H (FH), which inhibits alternative pathway activation. In the present study, using a mouse model of systemic infection and flow-cytometric analyses, we demonstrated an in vivo interaction between FH and pneumococci and showed differential FH binding during bacteremia. Flow-cytometric analyses of pneumococci harvested after intraperitoneal (ip) challenge demonstrated increased binding of FH, compared with that after intravenous (iv) challenge. Real-time polymerase chain reaction analyses of PspC mRNA showed that, relative to pneumococci grown in vitro, those recovered from the blood of mice 24 h after iv challenge exhibited 23-fold higher mRNA levels; however, after ip challenge, PspC mRNA induction was increased 870-fold. A subsequent increase in PspC expression was detected by flow cytometry using a monoclonal antibody against PspC. Furthermore, pneumococci with FH bound to complement before exposure had increased proliferation, compared with pneumococci not pretreated with FH. These results suggest that the interaction between PspC and FH contributes to pneumococcal virulence. PMID- 16267774 TI - A mathematical model quantifying the impact of antibiotic exposure and other interventions on the endemic prevalence of vancomycin-resistant enterococci. AB - BACKGROUND: Mathematical modeling can be used to describe the interdependent and dynamic interactions that contribute to the transmission dynamics of vancomycin resistant enterococci (VRE). A model was developed to quantify the contribution of antibiotic exposure and of other modifiable factors to the dissemination of VRE in the hospital setting. METHODS: The model consists of 4 compartments: patients colonized with VRE receiving and not receiving antibiotics and uncolonized patients receiving and not receiving antibiotics. A series of differential equations describe the movement between these compartments. Baseline parameter estimates were obtained from pharmacy, infection-control, and clinical databases. RESULTS: The main predictions of this model are that (1) preventing the initiation or enhancing the discontinuation of unnecessary antimicrobial therapy will have a greater impact if it is targeted to patients who are not colonized with VRE; (2) increasing the number of patients harboring VRE at the time of hospital admission substantially increases the endemic prevalence of VRE; and (3) eliminating the influx of VRE results in the eradication of this pathogen from the hospital. A decrease in the endemic prevalence of VRE also occurs with a decrease in the length of hospital stay of colonized patients, increased hand hygiene compliance, and a lower ratio of health-care workers : patients. CONCLUSION: This mathematical model provides a framework to assist in targeting necessary interventions aimed at limiting the spread of VRE. PMID- 16267775 TI - The role of epitope specificity in the human opsonic antibody response to the staphylococcal surface polysaccharide poly N-acetyl glucosamine. AB - BACKGROUND: The staphylococcal surface polysaccharide poly N-acetyl glucosamine (PNAG) is a target for killing and protective antibody in animals. We investigated the human antibody response and specificity of binding and opsonic antibodies for different epitopes on PNAG in serum samples from patients with cystic fibrosis (CF) colonized and not colonized with Staphylococcus aureus. METHODS: Serum samples from patients with CF colonized and not colonized with S. aureus were used to compare levels and specificities of binding and opsonic antibodies to native PNAG (>95% acetylation) and deacetylated PNAG (dPNAG, approximately 15% acetylation). RESULTS: Colonized patients had higher killing activity mediated by opsonic antibody than did noncolonized patients in a PNAG specific opsonophagocytic assay (P<.0001) but no difference in average levels of antibody to either PNAG or dPNAG by enzyme-linked immunosorbent assay. Killing activity in serum samples of the colonized patients correlated with the level of IgG specific to dPNAG more than to native PNAG. dPNAG and PNAG shared expression of the epitopes binding opsonic antibody, as evidenced by comparable inhibition of opsonophagocytic killing by both antigens. Affinity-purified antibodies specific to dPNAG were superior in mediating opsonic killing. CONCLUSION: Human antibodies to PNAG that mediate opsonic killing bind primarily to the nonacetylated epitopes of this antigen, which indicates that these antigenic determinants are the dominant targets of the functional human antibody response to staphylococcal PNAG. PMID- 16267776 TI - Association between human leukocyte antigen class II alleles and genotype of Borrelia burgdorferi in patients with early lyme disease. AB - BACKGROUND: On the basis of a polymerase chain reaction-restriction fragment length polymorphism analysis of the 16S-23S ribosomal DNA intergenic spacer, clinical isolates of Borrelia burgdorferi can be classified into 3 genotypes designated as RST1, RST2, and RST3. RST1 strains are the most pathogenic, and RST3 strains are the least pathogenic. METHODS: Human leukocyte antigen (HLA) class II alleles were determined for a group of culture-positive patients with Lyme disease-associated erythema migrans and were evaluated for an association with the genotype of the infecting B. burgdorferi strain. RESULTS: The DRB1*0101 allele carriage rate was higher in patients infected with RST3 strains (9/25 [36.0%]) than in patients infected with RST1 strains (2/28 [7.1%]) or RST2 strains (7/36 [19.4%]) (P=.010). The same relationship was found for carriage of the DRB1*0101-DQB1*0501 haplotype (P=.018), because of tight linkage disequilibrium. Similar associations could not be demonstrated for any of the other DRB1 and DQB1 alleles or haplotypes that were assessed. CONCLUSION: The DRB1*0101 allele and the DRB1*0101-DQB1*0501 haplotype may be relevant to the development of infection with strains from the least invasive genotypes of B. burgdorferi. PMID- 16267777 TI - Internationally recognized antimicrobial susceptibility testing methods and interpretive criteria: the case for conformity. PMID- 16267779 TI - Salmonella serotype typhimurium, not antimicrobial resistance per se, is associated with excess bloodstream infections and hospitalizations. PMID- 16267781 TI - Ghrelin cell density in the gastrointestinal tracts of animal models of human diabetes. AB - Ghrelin cell density in the gastrointestinal tract of animal models of human diabetes type 1 and 2 was investigated. The animals used were non-obese diabetic (NOD) mice and obese diabetic mice. Ghrelin cells were detected by immunohistochemistry and quantified by computerized image analysis. Ghrelin immunoreactive cells were detected in all animals studied. They were abundant in the oxyntic mucosa, patchy and few in the duodenum and rare in the colon. The density of ghrelin-immunoreactive cells decreased in diabetic, pre-diabetic NOD mice and in obese diabetic mice as compared to controls, though not statistically significant. It was concluded that the reduced density of ghrelin-immunoreactive cells in animal models of human diabetes type 1 and 2 might explain the slow gastric emptying and slow intestinal transit found in diabetes gastroenteropathy. PMID- 16267782 TI - Frequent expression of neuroendocrine markers in mucinous tubular and spindle cell carcinoma of the kidney. AB - Mucinous tubular and spindle cell carcinoma (MTSCC) is a new tumorous entity which has been recently established. In this article, we examined the expression of neuroendocrine markers including neuron specific enolase (NSE), chromogranin A and synaptophysin in 16 cases of MTSCC using immunohistochemistry. The sex ratio (male: female) of the patients was 4:12. In normal kidney, distal tubules or collecting ducts were positive for NSE, but no structures were positive for chromogranin A or synaptophysin. All MTSCCs showed a positive reaction for NSE. Additionally, fifteen of sixteen neoplasms (93.8%) with MTSCC showed the expression of either chromogranin A or synaptophysin or both. Finally, it is possible that MTSCC may be one of renal neoplasms which frequently exhibit the neuroendocrine differentiation. PMID- 16267783 TI - Histochemical analysis of glycoconjugates in the domestic cat testis. AB - The localization and characterization of oligosaccharide sequences in the cat testis was investigated using 12 lectins in combination with the beta-elimination reaction, N-Glycosidase F and sialidase digestion. Leydig cells expressed O linked glycans with terminal alphaGalNAc (HPA reactivity) and N-glycans with terminal/internal alphaMan (Con A affinity). The basement membrane showed terminal Neu5Acalpha2,6Gal/GalNAc, Galbeta1,3GalNAc, alpha/betaGalNAc, and GlcNAc (SNA, PNA, HPA, SBA, GSA II reactivity) in O-linked oligosaccharides, terminal Galbeta1,4GlcNAc (RCA120 staining) and alphaMan in N-linked oligosaccharides; in addition, terminal Neu5acalpha2,3Galbeta1,4GlcNac, Forssman pentasaccharide, alphaGal, alphaL-Fuc and internal GlcNAc (MAL II, DBA, GSA I-B4, UEA I, KOH sialidase-WGA affinity) formed both O- and N-linked oligosaccharides. The Sertoli cells cytoplasm contained terminal Neu5Ac-Galbeta1,4GlcNAc, Neu5Ac-betaGalNAc as well as internal GlcNAc in O-linked glycans, alphaMan in N-linked glycoproteins and terminal Neu5Acalpha2,6Gal/ GalNAc in both O- and N-linked oligosaccharides. Spermatogonia exhibited cytoplasmic N-linked glycoproteins with alphaMan residues. The spermatocytes cytoplasm expressed terminal Neu5Acalpha2,3Galbeta1,4 GlcNAc and Galbeta1,3GalNAc in O-linked oligosaccharides, terminal Galbeta1,4GlcNAc and alpha/betaGalNAc in N-linked glycoconjugates. The Golgi region showed terminal Neu5Acalpha2,3Galbeta1,4GlcNac, Galbeta1,4GlcNAc, Forssman pentasaccharide, and alphaGalNAc in O-linked oligosaccharides, alphaMan and terminal betaGal in N-linked oligosaccharides. The acrosomes of Golgi-phase spermatids expressed terminal Galbeta1,3GalNAc, Galbeta1,4GlcNAc, Forssmann pentasaccharide, alpha/betaGalNAc, alphaGal and internal GlcNAc in O-linked oligosaccharides, terminal alpha/betaGalNAc, alphaGal and terminal/internal alphaMan in N-linked glycoproteins. The acrosomes of cap-phase spermatids lacked internal Forssman pentasaccharide and alphaGal, while having increased alpha/betaGalNAc. The acrosomes of elongated spermatids did not show terminal Galbeta1,3GalNAc, displayed terminal Galbeta1,4GlcNAc and alpha/betaGalNAc in N glycans and Neu5Ac-Galbeta1,3GalNAc in O-linked oligosaccharides. PMID- 16267784 TI - Altered alpha1-syntrophin expression in myofibers with Duchenne and Fukuyama muscular dystrophies. AB - Alpha1-syntrophin, a scaffolding adapter and modular protein, is a cytoplasmic component of the dystrophin glycoprotein complex. This study investigated immunohistochemically the expression of alpha1-syntrophin in Duchenne and Fukuyama muscular dystrophies (DMD and FCMD, respectively). Biopsied muscles of five DMD, five FCMD, five normal controls and five disease controls (three myotonic and two facioscapulohumeral dystrophies) were analyzed. Immunoblot analysis showed that anti-alpha1-syntrophin antibody had a decreased reaction in both DMD and FCMD muscle extracts. Biopsied muscle sections and their serial sections were immunostained with rabbit anti-alpha1-syntrophin and rabbit anti muscle-specific beta-spectrin antibodies, respectively. Immunoreactive patterns of sarcolemma were classified into (i) a continuously positive immunostaining pattern, (ii) a partially positive immunostaining pattern, (iii) a negative immunostaining pattern and (iv) a faint but entire surface positive immunostaining pattern. The group mean percentages of alpha1-syntrophin and beta spectrin immunonegative myofibers in the DMD group were 39.3% and 10.8%, respectively, while those in the FCMD group were 45.5% and 10.4%, respectively. These values were statistically significant compared with those of disease control and normal control muscles. Thus we found that dystrophin-deficient DMD muscles contained significant numbers of alpha1-syntrophin-positive fibers and significant numbers of alpha1-syntrophin-negative fibers were present in dystrophin-positive muscles of severe muscular dystrophy such as FCMD. Alpha dystrobrevin immunoreactivity was tested in DMD muscles and appreciable amounts of alpha-dystrobrevin that binds to syntrophin were found in DMD muscle membranes. PMID- 16267785 TI - Immunohistochemical study of serum albumin in normal and cadmium-treated mouse testis organs by "in vivo cryotechnique". AB - The in vivo injection of cadmium (Cd) was reported to induce blood-testis barrier disruption, and assumed to be an experimental model to examine junctional structures in seminiferous tubules. The purpose of this study is to investigate time-dependent changes of albumin permeability in the normal or Cd-treated mouse testis by our "in vivo cryotechnique" with immunohistochemistry, reflecting tight junctional (TJ) barriers of Sertoli cells. The albumin in the seminiferous tubules was firstly immobilized by the cryotechnique, in which normal blood circulation was always kept. The cryofixed testicular tissues were then processed for freeze-substitution, and embedded in the paraffin wax. Serial sections were immunostained by anti-mouse albumin antibody with peroxidase immunostaining, and also stained with hematoxylin-eosine (HE) for morphological observation. In normal seminiferous tubules, the immunoreaction products were localized around peritubular myoid cells and between Leydig cells, as well as in blood vessels. They were also localized as arch-like patterns around some spermatogonia in basal compartments of seminiferous tubules. Twenty-four and 48 hrs after Cd-treatment, some enlarged spaces and vesicular formations in the seminiferous epithelium were observed on the HE-stained sections. The albumin immunolocalization was detected not only in the basal compartments, but also in the adluminal compartments between Sertoli cells and germ cells. Thus, the structural disruptions of inter Sertoli TJ barriers could be clearly demonstrated by the "in vivo cryotechnique". PMID- 16267786 TI - Investigation of general and cytoskeletal markers to estimate numbers and proportions of neurons in the human intestine. AB - An important requirement in pathological diagnostics in the human enteric nervous system (ENS) is the estimation of the total numbers of neurons and of proportions of distinct subpopulations. In this study, we compared the suitability of two suggested panneuronal markers, cuprolinic blue (CB) and anti-Hu-protein (HU), for staining and counting human myenteric neurons in wholemounts, derived from small and large intestinal samples. Furthermore, the proportional expression of three cytoskeletal intermediate filaments, alpha-internexin (IN), neurofilament 200 (NF) and peripherin (PE), was correlated with both CB and HU. In 8 CB- and HU stained wholemounts, 93.3% of all neurons were double labeled, 3.3% of neurons were stained only with CB whereas 3.3% were immuno-stained only for HU. Thus, both markers were comparably reliable in representing the putative total human myenteric neuron population in our material. The wholemounts double stained for IN/CB or IN/HU revealed between 56.2 and 71.5% of neurons to be IN-reactive. Between 42.8 and 50.9% of neurons were immunoreactive for NF whereas 53.9 to 62.4% of neurons were reactive for PE. Although our sample number was too small to allow final conclusions, we suggest that the variations in proportions of intermediate filament expression we observed may be due to individual circumstances rather than to correlation with age or region. The proportions of neurons positive for IN, NF or PE but unstained by CB histochemical or HU immunohistochemical techniques was between 0 and 2.2%. We conclude that both CB and HU techniques are suitable methods for representation of almost all myenteric neurons in the human gut and that the differential expression of the cytoskeletal proteins investigated has to be included in the classification of enteric neurons in pathological diagnostics of human gastrointestinal diseases. PMID- 16267787 TI - The penis: a new target and source of estrogen in male reproduction. AB - In the past decade, interest and knowledge in the role of estrogen in male reproduction and fertility has gained significant momentum. More recently, the cellular distribution and activity of estrogen receptors (alpha and beta)(ER) and aromatase (estrogen synthesis) has been reported in the penis, making the penis the latest "frontier" in the study of estrogen in male reproduction. ER and aromatase are broadly and abundantly expressed in various penile compartments and cell types (erectile tissues, urethral epithelia, vascular and neuronal cells), suggesting the complexity and significance of the estrogen-ER system in penile events. Unraveling this complexity is important and will require utilization of the various resources that are now at our disposal including, animal models and human lacking or deficient in ER and aromatase and the use of advanced and sensitive techniques. Some of the obvious areas that require our attention include: 1) a comprehensive mapping of ER-alpha and -beta cellular expression in the different penile compartments and subpopulations of cells, 2) delineation of the specific roles of estrogen in the different subpopulations of cells, 3) establishing the relationship of the estrogen-ER system with the androgen androgen receptor system, if any, and 4) characterizing the specific penile phenotypes in human and animals lacking or deficient in estrogen and ER. Some data generated thus far, although preliminary, appear to challenge the long held dogma that, overall, androgens have a regulatory monopoly of penile development and function. PMID- 16267788 TI - The NF-kappaB-mediated control of ROS and JNK signaling. AB - NF-kappaB/Rel transcription factors are best known for their roles in innate and adaptive immunity and inflammation. They also play a central role in promoting cell survival. This latter activity of NF-kappaB antagonizes programmed cell death (PCD) induced by the proinflammatory cytokine tumor necrosis factor (TNF)alpha and plays an important role in immunity, lymphopoiesis, osteogenesis, tumorigenesis and radio- and chemoresistance in cancer. With regard to TNFalpha, the NF-kappaB-mediated inhibition of PCD seems to involve an attenuation of the c Jun-N-terminal kinase (JNK) cascade mediated through the induction of select downstream targets such as the caspase inhibitor XIAP, the zinc-finger protein A20, and the inhibitor of the MKK7/JNKK2 kinase, Gadd45beta/Myd118. Notably, NF kappaB also blunts accumulation of reactive oxygen species (ROS), which themselves are pivotal elements for induction of PCD by TNFalpha, and this suppression of ROS formation mediates an additional protective activity recently ascribed to NF-kappaB. The antioxidant activity of NF-kappaB has been shown to depend upon upregulation of both Ferritin heavy chain (FHC)--a component of Ferritin, the primary iron-storage protein complex found in cells--and of the mitochondrial enzyme Mn++ superoxide dismutase (Mn-SOD). Indeed, the inductions of Mn-SOD and FHC represent another important means through which NF-kappaB controls proapoptotic JNK signaling triggered by TNFalpha. These findings might enable the development of new, more targeted approaches to treatment of diseases sustained by a deregulated activity of NF-kappaB, including some cancers and chronic inflammatory conditions. PMID- 16267789 TI - Current concepts in ovarian epithelial tumorigenesis: correlation between morphological and molecular data. AB - Ovarian carcinoma is the most lethal gynaecological malignancy, most tumours being advanced at presentation. However, little is known about precursor lesions and the cell of origin of epithelial ovarian malignancy. In this review, the proposed cell of origin is discussed as well as recent molecular data relating to ovarian cancers of different morphological types. It is stressed that ovarian carcinoma is a heterogeneous group of neoplasms with several different morphological types, each with their own underlying molecular genetic events. Recent data suggest that mucinous ovarian cancers and a small subset of serous cancers (low grade ovarian serous carcinoma) develop through a well-defined adenoma-carcinoma sequence while the much more common high grade ovarian serous carcinoma develops de novo from the ovarian surface epithelium or the epithelium of cortical inclusion cysts. The realisation that various morphological types of epithelial ovarian cancer are associated with different molecular genetic events is a major advance in the study of ovarian cancer. It can be anticipated that this will lead to the development of specific therapeutic agents of value against a specific tumour type. PMID- 16267790 TI - Recombinant generation of two fragments of the rat complement inhibitory factor H [FH(SCR1-7) and FH(SCR1-4)] and their structural and functional characterization in comparison to FH isolated from rat serum. AB - Factor H (FH) is the predominant soluble inhibitor of the complement system. With a concentration of 200-800 microg/ml in human and rat plasma it acts as a cofactor for the soluble factor I (FI)-mediated cleavage of the component C3b to iC3b. Furthermore it competes with factor B for binding to C3b and C3(H2O) and promotes the dissociation of the C3bBb complex. FH is a monomer of about 155 kDa which comprises 20 short consensus repeats (SCR), each of which is composed of approximately 60 amino acid (aa) residues. Two functional fragments of FH comprising the SCR1-4 or SCR1-7 were generated using either the Baculovirus system or stably transfected human embryonal kidney cells, respectively. These fragments, as well as FH purified from rat serum, were first analyzed for their relative molecular weights (Mr) using non-reducing or reducing SDS-PAGE. The Mr of the FH variants differed by about 20% depending on the experimental conditions employed. Only the Mr of proteins separated under reducing conditions were in accordance with the MW calculated from the aa sequence. Analyses of the glycosylation patterns using PAS-staining showed a lack of staining of the recombinant variants (SCR1-4 and SCR1-7) in contrast to FH(SCR1-20) from serum. Using a complement hemolysis assay (CH50-assay) all three variants exhibited a molar complement inhibitory activity of FH(1-20)/FH(1-7)/FH(1-4) of about 3/1/1. These data support the postulated model of FH bearing three binding sites for its ligand C3b, from which one is located in the SCR1-4, whereas the other two are located in the SCR8-20. PMID- 16267793 TI - [New insights in the pathogenesis of secondary hyperparathyroidism]. AB - Parathyroid gland growth is a major cause of secondary hyperparathyroidism in renal failure. It is well known that high serum phosphate levels, low serum calcium levels and vitamin D deficiency are the three promoters of parathyroid hyperplasia in renal failure. Recent studies have investigated in depth the potential role of growth factors (transforming growth factor alpha) and their receptors (epidermal growth factor receptor) in the pathogenesis of parathyroid cell hyperplasia in chronic renal failure. The identification of molecular mechanisms involved in calcium, phosphate and vitamin D manipulations in an experimental renal failure model could help design more effective therapy for secondary hyperparathyroidism in uremic patients. PMID- 16267792 TI - [Use of dialysate conductivity to obtain neutral sodium balance in chronic hemodialysis patients]. AB - In order to assure a zero sodium balance in hemodialysis patients, attaining 'constant' values of total body water and plasma water sodium concentration at the end of each dialysis session is a basic pre-requisite. This is achieved by matching the ultrafiltration to the inter-dialytic weight gain and by individualizing dialysate sodium concentration at each dialysis session by making use of a kinetic model. Clinical results suggest that the single pool variable volume sodium kinetic model allows the targeted end-dialysis plasma water sodium concentration to be obtained. Nevertheless, this model is not suitable for routine clinical application, because of difficulties in the real-time determination of initial plasma water sodium concentration and 'effective' sodium dialysance. Measuring dialysate conductivity at the inlet and outlet ports of the dialyzer allows the estimation of sodium transfer during dialysis, if the function of concentration versus conductivity is known. If sodium transfer is measured at two different inlet dialysate conductivities, it is possible to determine ionic dialysance and systemic plasma water conductivity, which can be used routinely to apply the single pool sodium kinetic model. Given that ionic dialysance and plasma water conductivity can be measured easily repeatedly and inexpensively at each dialysis session without the need for blood sampling or laboratory determinations, it can be expected that conductivity kinetic models will soon become a part of everyday clinical practice. PMID- 16267795 TI - [Statistical models and multivariable analysis]. AB - Most clinical research can be simplified as an investigation of an input/output relationship. The inputs are called explanatory (independent) variables or predictors and are thought to be related to the outcome, or response (independent) variable. This relationship is usually complicated by other factors related to both the input and the output (presence of confounding) and can vary according to the levels of the other variables (presence of interaction). This input/output relationship is usually described by statistical models that include a fit part and a residual component or difference between the data and the fit. The most popular models are the general linear models, which can be considered the paradigm of all models used in multi-variable analyzes. PMID- 16267794 TI - [Kidney transplantation in children]. AB - Indications, procedures, complications, pharmacokinetics and outcomes of renal transplantation are different in children and in adults. Subjects <18 yrs old, are often included in a unique list as in Italy, benefiting from donors <15 yrs old, and the waiting time is reduced to <12 months in 71% of cases. The risk of thrombosis limits the use of donors <2 yrs and trans-plantation in infants <1 yr. The age at kidney transplantation is <5 yrs in 20-30% of children. In Italy living-related trans-plantation (LRT) is performed in 7% of cases, while in the USA it is more common (57%) and is often pre-emptive before entering dialysis (24%). Current therapy tends to reduce steroid treatment doses and, optimizing induction therapy with IL-2R inhibitors, using tacrolimus or mycophenolate or sirolimus. Transplanted patient survival is better in children than in adults (94 98% at 5 yrs). Infections, cardiovascular diseases and neoplasia induce 34, 15 and 12% of deaths, respectively, at 10 yrs; morbidity for infections and lymphoproliferative disease is increasing. Acute rejections declined from 70% in 1987 to 31% in 2002 in cadaveric transplantation (CT) and renal survival at 3 yrs increased from 50% in 1985 to 82% for CT and up to 92% in LRT. In adolescents (11 17 yrs old) renal survival is lower than in infants and in adults <65 yrs old. Renal losses are due to chronic transplant nephropathy (32%), vascular thrombosis (13%) and the recurrence of the original nephropathy (focal glomerulosclerosis up to 50%, membrano-proliferative glomerulonephritis up to 30%, and primary hyperoxaluria up to 90% if combined kidney-liver transplantation is not performed). Growth improves after transplantation particularly in children <5 yrs, while it is not completely satisfactory in adolescents. Overall, results indicate that kidney transplantation in children has very much improved and will offer in the near future even more favorable outcomes. PMID- 16267797 TI - [Dialytic treatment in patients with renal insufficiency and heart failure refractory to combined drug therapy]. AB - The incidence and prevalence of heart failure are continuously on the increase. Pharmacological therapy is not sufficient in the advanced stages of the illness to control the signs and symptoms, especially when stressing factors intervene and complicate the illness course. Dialytic treatments are always indicated more than necessary in the severe forms of heart failure, but the optimal treatment has not yet been established. In our division, 24 patients with heart failure (III-IV NYHA) and renal failure (III-IV NKF-DOQI) were treated with low flux bicarbonate dialysis, variable dialysate and limited ultra-filtration. Arrhythmia, pericarditis, ischemic cardiopathy and hyponatremia were the stressing factors that determined acute heart failure. Fifteen patients presented with diastolic failure, whereas nine patients had systolic heart failure. The first group of patients had higher arterial pressure, better ejection fraction and better prognosis regarding renal function and survival rate. The results obtained with this treatment in prevalently diastolic heart failure are satisfactory in terms of survival, as well as in renal function recovery, whereas they are not adequate in systolic failure where other approaches are being studied. Therefore, we believe that the therapeutic choice must always be determined in relation to the type of heart failure. PMID- 16267796 TI - [Census 2004 of the Italian Renal and Dialysis Units--Piemonte, Liguria and Valle d'Aosta]. AB - The Italian Society of Nephrology (SIN) promoted a national survey in order to obtain detailed information from all Renal and/or Dialysis Units using the on line questionnaire (158 items) regarding structural and technological resources, medical workforce organisation and activity features. The purposes of this initiative were to obtain regional benchmarks as references for renal units and to describe the current Italian renal network in order to plan further interventions for the next 5 years. In this paper data of the first three Italian Regions (Piemonte, Liguria and Valle d'Aosta) which completed the survey (100% of the units) are reported. Main findings in the 3 Regions. A) Epidemiology: prevalence of dialysis patients = 709, 720, 787 pmp (per million population); prevalence of transplanted patients = 325, 387, 279 pmp; incidence of dialysis patients = 166, 191, 156 pmp; gross mortality of dialysis patients = 13.7, 15.0, 13.0%; distribution of vascular access in prevalent dialysis patients: arteriovenous fistula = 74, 83, 76%, central venous catheter = 18, 12, 15%, vascular graft = 8, 5, 9%. B) Structural resources: hospital's number of beds = 49, 72, 49 pmp, dialysis places = 166, 158, 164 pmp. C) Personnel resources: renal physicians = 44, 47, 41 pmp, renal nurses = 186, 194, 205 pmp; each renal physician takes care of 16, 15, 19 dialysis patients and each renal nurse cares for 3.8, 3.7, 3.8 dialysis patients. D) Activity: admission to hospital = 1507, 2392, 1606 pmp, renal biopsies = 109, 133, 57 pmp. Despite discrepancies in population density in the three Regions, most indexes are surprisingly similar and show the satisfactory level of renal care attained in the Northwestern Italian area. Further improvements in health care management can be predicted as a consequence of a direct comparison between needs and results in the various Regions of the Country. PMID- 16267798 TI - [A 'pretty' surprise...]. AB - This is a clinical history of hypertensive chronic renal failure patients who first presented at our hypertension ambulatory for control. The possibility of performing routine echography permitted the precocious diagnosis of bilateral kidney neoplasm and urothelial cancer. PMID- 16267799 TI - [From the Mailing List SIN: expected and unexpected professional risks for the nephrologists--reflections from an outbreak of Burkholderia cepacia bacteremia in a hemodialysis unit]. AB - An outbreak of bacteremia in 20 hemodialysed patients who developed central venous catheter (CVC) infection related to Burkholderia cepacia is reported, introducing medical and professional responsibilities in nephrology units. The cepacia was documented in the blood stream, in the CVC biofilm, in the water supply and in the distribution. This and other confounding factors delayed the identification of the contamination source. Finally, it was isolated, clonally identical to that found in the blood stream, from ammonium chloride solution used to disinfect the skin and distributed in a sterile disposable kit. Burkholderia cepacia was clonally different in blood with respect to water. The possible differing responsibilities in the organizational steps of nephrology activity are discussed. PMID- 16267801 TI - [Bioelectrical analysis in uremic patients]. AB - Bioelectrical analysis (BIA) is an easy, repeatable, low cost, operator independent method. BIA obtains two different goals, i.e. body water content evaluation, by the RXc Graph or the BIVA Z score and morbidity and mortality predictions by the phase angle. Therefore, BIA can be considered as part of the clinical examination for the evaluation of both hydration and nutritional status. PMID- 16267800 TI - [Fabry nephropathy in a female with superposed IgA glomerulonephritis]. AB - BACKGROUND: In Anderson-Fabry disease (AFd), the kidney is affected in all hemizygous males and in some heterozygous females. Female carriers can present subtle renal abnormalities due to glycosphingolipid (GSL) accumulation within renal cells. Renal biopsy is rarely performed in female Fabry patients because clinical renal manifestations are usually lacking. However, female carriers can accumulate GSL in their renal cells despite the absence of clinically evident kidney disease. CASE REPORT: We performed a kidney biopsy in a 52-year-old female patient, a Fabry disease carrier. The patient showed normal glomerular filtration rate, persistent microhematuria and proteinuria (about 1.7 g/24 hr), cornea "verticillata", and evident left ventricular hypertrophy. The molecular study documented a missense mutation R227Q in exon 5 of the alpha-galactosidase A gene. Optical microscopy showed electron-dense mesangial deposits due IgA glomerulonephritis, as confirmed by immunofluorescence. We decided to start therapy with angiotensin-converting enzyme inhibitors (ACE-I). After 8 months of treatment, the patient demonstrated proteinuria of 0.9 g/24 hr. To decide when to start treatment using enzyme replacement therapy (ERT) with human recombinant GAL A (Fabrazyme), we decided to perform an electron microscopy study of the renal biopsy. The renal ultrastructural findings were typical GSL inclusions in all kinds of glomerular cells, in tubular epithelial cells and in endothelial cells of interstitial capillaries, confirming the hypothesis of Fabry nephropathy. Consequently, Fabrazyme was given at a standard dose of 1 mg/kg every 2 weeks. After 24 months of combined treatment (ACE-I-Fabrazyme), proteinuria decreased to 0.2 g/24 hr. CONCLUSIONS: The importance of performing the ultrastructural examination of the kidney biopsy is stressed, especially in heterozygous Fabry patients to evaluate the need to treat them with ERT and to evaluate the degree of renal involvement. PMID- 16267791 TI - Winding through the WNT pathway during cellular development and demise. AB - In slightly over a period of twenty years, our comprehension of the cellular and molecular mechanisms that govern the Wnt signaling pathway continue to unfold. The Wnt proteins were initially implicated in viral carcinogenesis experiments associated with mammary tumors, but since this period investigations focusing on the Wnt pathways and their transmembrane receptors termed Frizzled have been advanced to demonstrate the critical nature of Wnt for the development of a variety of cell populations as well as the potential of the Wnt pathway to avert apoptotic injury. In particular, Wnt signaling plays a significant role in both the cardiovascular and nervous systems during embryonic cell patterning, proliferation, differentiation, and orientation. Furthermore, modulation of Wnt signaling under specific cellular influences can either promote or prevent the early and late stages of apoptotic cellular injury in neurons, endothelial cells, vascular smooth muscle cells, and cardiomyocytes. A number of downstream signal transduction pathways can mediate the biological response of the Wnt proteins that include Dishevelled, beta-catenin, intracellular calcium, protein kinase C, Akt, and glycogen synthase kinase-3beta. Interestingly, these cellular cascades of the Wnt-Frizzled pathways can participate in several neurodegenerative, vascular, and cardiac disorders and may be closely integrated with the function of trophic factors. Identification of the critical elements that modulate the Wnt Frizzled signaling pathway should continue to unlock the potential of Wnt pathway for the development of new therapeutic options against neurodegenerative and vascular diseases. PMID- 16267802 TI - [Improving the outcome of peritoneal dialysis in the long term: is it possible?]. AB - Improving the results of peritoneal dialysis (PD) over time means reducing both the technique's drop out (TDO) and mortality rates. The PD mortality rate has diminished over the years, due to greater experience in using the technique and the reduction in mortality due to peritonitis making it comparable with the hemodialysis (HD) mortality rate. Moreover, improved control of the hydrosaline balance through the use of ambulatory peritoneal dialysis (APD) and icodextrin could further improve the survival rate in the future. The adequacy targets needed to reduce the mortality rate still appear to be debatable, as their importance seems conditioned by the presence of RRF and comorbidity. The TDO is higher in PD than in HD because PD is a self-administered treatment that uses a biological membrane as a filter. The most frequent causes of TDO are peritonitis (30-40%), dialytic inadequacy (11-27%), and subsequent inability and/or choice (10-32%). Peritonitis is the cause that has seen the greatest reduction over time due to the introduction of the Y-Set, but a further reduction could result from the prevention of ESI, and from improvements in the patient selection procedure designed to identify both clinical and psycho-social disposition peritonitis risk factors. Among the causes of TDO due to dialytic inadequacy, insufficient ultrafiltration (UF) could benefit from the diffusion of APD and icodextrin, while insufficient depuration could be reduced by new targets and optimization of the prescription. Finally, TDO due to social causes could be reduced by the use of APD, care support and appropriate patient selection. PMID- 16267803 TI - [Hypertension in patients on hemodialysis: the role of salt intake]. AB - In the 1960s, about 10% of hemodialysis (HD) patients had hypertension; the current percentage of hypertensive patients has risen to 70-75%. The scarce implementation of low-salt diets and the increment of dialysate sodium concentration aimed at ameliorating treatment tolerability are the main causes of the currently poor hypertension control. Considerable sodium intake activates a vicious circle: an increase in serum osmolarity, greater thirst and greater water intake, high inter-dialytic weight gains, need for large ultrafiltration rates, more frequent episodes of intradialytic hypotension, failure to achieve dry weight, progressive extra-cellular volume (ECV) expansion, and finally, blood pressure (BP) increase. Therefore, many studies have pointed out the importance of a low-salt diet in HD; it has been proven that the normalization of BP and ECV overload with a low-salt diet is associated with left ventricular hypertrophy regression and diastolic dysfunction improvement. Preparing meals with fresh foods, using spices, avoiding salt when cooking, and drastically limiting salty foods reduce dietary sodium down to about 6 g/day. Sodium intake during inter dialytic periods can easily be assessed by measuring the changes in serum sodium concentration and in body weight. PMID- 16267804 TI - [Clinical and genetic features of the Alport 'syndromes']. AB - Alport syndrome (ATS) is a clinically and genetically heterogeneous progressive nephropathy often associated with deafness and/or ocular lesions. The histological aspect is characterized by thinning, thickening and splitting of the glomerular basement membrane (GBM). Alport syndrome is caused by mutations in COL4A3 gene (type IV collagen, alfa-3 chain), or COL4A4 gene (type IV collagen, alfa-4 chain) or COL4A5 gene (type IV collagen, alfa-5 chain) genes. Alport syndrome accounts for 1-2% of renal failure cases in Europe, and for 2-3% of transplanted patients in United States. This review focuses on the three types of Alport syndrome which differ in the clinical progression and in the mode of inheritance. The common X-linked form is caused by mutations in the COL4A5 gene and it accounts for 85% of cases. The autosomal dominant and the autosomal recessive forms are caused by mutations in either COL4A3 or COL4A4 genes. The autosomal recessive form which is responsible for the 10-15% of Alport cases, has been known since several years. On the contrary, the autosomal dominant form has only recently been identified in some families. Furthermore, this review will focus on the difficulties encountered during the genetic counselling related to the differential diagnosis between Alport syndrome and Thin Basement Membrane Disease (TBMD). We will report direct experiences of our group showing the difficulties to give an exact prognosis and a correct recurrence risk to the family. PMID- 16267805 TI - [Health related quality of life (HRQoL) assessment in uremic patients: a qualitative review of concepts, methods, available instruments and results]. AB - The health related quality of life (HRQoL) assessment is a complex and multidimensional concept usually defined as the perception by individuals of the impact of diseases and medical treatments on physical, psychological and social aspects of health and life. HRQoL has become more popular in the last 20 yrs as an important indicator of the effectiveness of the medical care in clinical research protocols, and as part of so-called patient-reported outcomes (PRO), which are concepts concerning aspects of health perceived, reported and rated by study subjects or patients. Generic and disease specific questionnaires are usually used to assess the individual's perceptions about health and life, and more valid and reliable instruments are now available to use in research settings. Instruments that can be useful in daily clinical practice, in the management of the single patient, are not yet available. A large portion of the scientific literature dealing with HRQoL is actually focused on many aspects of uremic syndrome, renal transplantation and dialysis therapies, probably because several reasons make this clinical setting particularly favorable for HRQoL assessment. Early referral to a nephrologist, anemia treatment with epoetin, daily and nocturnal dialysis, kidney transplantation, a program of physical exercise and the treatment of some co-existent conditions and symptoms (i.e. depression, pain, erectile dysfunction and restless leg symptoms) are the most effective ways to ameliorate HRQoL in renal patients. Moreover, HRQoL is a reliable prognostic factor for morbidity and mortality in uremic patients. PMID- 16267806 TI - [Introduction of the general linear models]. AB - General linear models can be considered the paradigm of all models used in clinical epidemiology. In these models, the independent variables combine in linear fashion to predict the values of the variable response. Since no model predicts the variable response perfectly, an error term is incorporated into the model to acknowledge what remains to be explained after getting a fit to the data. When this error term is normally distributed with constant variance, the linear models are reasonably appropriate to describe the input/output relationship of interest. PMID- 16267808 TI - [Twinkling artifact in kidney stone disease]. AB - BACKGROUND: Ultrasonography artifacts are false representations of the image caused by the interaction between the ultrasound and the tissues. The ability to identify artifacts is an important source of information that can help the clinician in performing a correct diagnosis. The twinkling artifact (called 'Effetto Arlecchino' by the Italians) consists of a series of colored pixels that appear inside, around and often along the shadow cone of the calculi. METHODS: We evaluated the clinical effectiveness of this artifact in the diagnosis of kidney stone disease. In 107 ultrasonography cases of hyperechogenic formations with clinical features of kidney stones, we used the color box to evoke the twinkling artifact. Of the 107 cases, 102 cases (95%) presented this artifact, while five cases did not. Moreover, this artifact was present in all urethral and bladder stones and in 62/67 kidney stone cases. CONCLUSIONS: In our experience, we found that the twinkling artifact is often positive in urinary stone disease, and the ability to identify it adds useful information to the diagnosis of urinary kidney stone disease. PMID- 16267809 TI - [From the Mailing List SIN: Epidemic of infections caused by 'aquatic' bacteria in patients undergoing hemodialysis via central venous catheters]. AB - Bacteremia due to central venous catheter (CVC) infection is the most frequent complication of CVC use as vascular access for hemodialysis (HD). We report a case of an epidemic of CVC infections caused by 3 strains of unusual bacteria: Ralstonia pickettii (Rp), Leifsonia xyli/Leifsonia aquatica (Lxa), Tsuckamurella strandjordae (Ts). From 20/8/01 to 30/9/01, 23 of 35 patients dialyzed via CVCs experienced intra-HD pyrogenic reactions. Their hemocultures were positive for: Rp (14 pts), Lxa (3 pts), Rp+Lxa (5 pts) and Rp+Ts (1 pt). The hemocultures of 12/35 asymptomatic pts were positive for: Rp 2 pts, Lxa 2 pts, Rp+Lxa 2 pts, Ts 1 pt, Rp+Ts 1 pt. The epidemiological and microbiological analyses of environmental samples failed to identify the source of the epidemic. Actions taken were: a) replacement of the batches of disposable materials; b) removal of CVCs in cases where possible to prepare a different access; c) treatment of the infections with intra-CVC antibiotic lock therapy. No relapses were recorded until April 2002, when 8 pts had again pyrogenic reactions due to Rp. After quick substitution of the CVC and repetitions of the action a), no relapses of pyrogenic reactions were observed. CONCLUSIONS: 1) given the characteristics of Rp, Lxa and Ts, saprophytes of moist environments, the most plausible source of the epidemic was a low-charge contaminated solution that was not identified due to low sensitivity of environmental sample culturing methods; 2) antibiotic lock therapy is a viable option for the conservative treatment of CVC infections. PMID- 16267807 TI - [The Dialysis Outcomes and Practice Patterns Study (DOPPS): results of the Italian cohort]. AB - BACKGROUND: The Dialysis Outcomes and Practice Patterns Study (DOPPS) is an international prospective, longitudinal, observational study examining the relationship between dialysis unit practices and outcomes for hemodialysis (HD) patients in seven developed countries France, Germany, Italy, Spain, United Kingdom, Japan and the United States. Results of the DOPPS in Italy are the subject of this report. METHODS: A national representative sample of 20 dialysis units (21 in Germany) was randomly selected in each of the European DOPPS countries (Euro-DOPPS). In these units, the HD in-center patients were included on a facility census, and their survival rates continuously monitored. A representative sample of incident (269 in Italy, 1553 in the Euro-DOPPS) and prevalent (600 in Italy, 3038 in the Euro-DOPPS) patients was randomly selected from the census for more detailed longitudinal investigation with regard to medical history, laboratory values and hospital admission. RESULTS: Comparing the Italian and Euro-DOPPS cohorts we found comparable mean age for prevalent patients (61.4 vs. 59.5 yrs), but incident patients were older in Italy. Italian prevalent patients had less cardiovascular disease, more satisfactory nutritional status and more frequent use of native vascular access. These data were associated with a comparable mortality (15.7 vs. 16.3 deaths/100 patient yrs), but morbidity was lower in Italy. Kt/V levels were comparable in the two cohorts (1.32 vs. 1.37), but 35% of Italian patients showed a Kt/V below the recommended target. Moreover, hemoglobin levels were below 11 g/dL in 60% of Italian patients. CONCLUSIONS: The DOPPS results bring to light several positive aspects and the opportunity for further possible improvements for Italian patients, but at the same time highlight some critical points that could represent a risk for dialysis quality. PMID- 16267810 TI - [Acute renal failure, hemolysis and thrombocytopenia]. AB - A 41-year-old male was admitted because of severe systemic hypertension and acute renal failure (ARF) that required hemodialysis (HD). Also present were hemolytic anemia, thrombocytopenia and increased plasmatic levels of aldosterone and reninic activity. The diagnostic tests performed during the recovery led to the conclusion of malignant hypertension. This case dealt with a cause of ARF, which is not currently so common; physicians should be aware of this condition especially when it is present with hemolytic anemia and thrombocytopenia, which are the microangiopathy markers. PMID- 16267811 TI - Blood and lymph vessels in embryonic tumors. AB - Formation of embryonic blood and lymph vessels is mediated by different steps of vasculogenesis and angiogenesis. The vascular endothelial growth factor family (VEGF) as well as the VEGF-receptors on the surface of blood endothelial cells and lymph endothelial cells are responsible for both processes. In addition to the embryonic development of the vessel systems, the interactions of angiogenesis factors and receptors are also present in solid tumors. Embryonic tumors in children offer an interesting target for new substances developed for anti angiogenesis. Some data on the role of blood vessels and anti-blood-angiogenesis are available on embryonic tumors. However, studies of lymph-angiogenesis are not found and anti-lymph-angiogenesis is not at all examined in embryonic tumors. This review gives an overview of the challenging field of angiogenesis and anti angiogenesis of both blood and lymph vessels with a focus on embryonic tumors. PMID- 16267812 TI - Systematic comparison of surface coatings for protein microarrays. AB - To process large numbers of samples in parallel is one potential of protein microarrays for research and diagnostics. However, the application of protein arrays is currently hampered by the lack of comprehensive technological knowledge about the suitability of 2-D and 3-D slide surface coatings. We have performed a systematic study to analyze how both surface types perform in combination with different fluorescent dyes to generate significant and reproducible data. In total, we analyzed more than 100 slides containing 1152 spots each. Slides were probed against different monoclonal antibodies (mAbs) and recombinant fusion proteins. We found two surface coatings to be most suitable for protein and antibody (Ab) immobilization. These were further subjected to quantitative analyses by evaluating intraslide and slide-to-slide reproducibilities, and the linear range of target detection. In summary, we demonstrate that only suitable combinations of surface and fluorescent dyes allow the generation of highly reproducible data. PMID- 16267813 TI - Proteome modifications of blue mussel (Mytilus edulis L.) gills as an effect of water pollution. AB - The discharge of chemicals such as oil associated or not with derived products constitutes a real threat for the environment. We report here the differential expression of the blue mussel (Mytilus edulis) gill proteins corresponding to two contaminated environmental conditions: crude oil and offshore produced water. In order to evaluate and understand contaminants, effects and adaptive response of these organisms, we identified proteins using MS. The latter can be grouped into three main classes: proteins involved in the cellular structure, in metabolism, and in defence proteins. PMID- 16267814 TI - Survey of albumin purification methods for the analysis of albumin-organic toxicant adducts by liquid chromatography-electrospray ionization-tandem mass spectrometry. AB - HSA has been shown to react with many organic toxicants to form adducts that are useful biomarkers for exposure. Albumin isolation is an important first step for the analysis of these protein-toxicant adducts. We tested several approaches to isolate albumin from serum treated with an electrophilic organic toxicant known to form adducts with albumin, i.e., sulfur mustard agent (HD) (2,2'-dichloroethyl sulfide), in order to evaluate these techniques as purification methods. To select the most efficient isolation strategy, methods were evaluated using gel electrophoresis, total protein quantitation, and peptide-adduct identification by MS. Results suggest that the albumin-rich fractions obtained can be used to identify exposure by quantitating the albumin adducts to electrophilic organic toxicants such as HD. The HiTrap Blue HP albumin isolation system appears to display the most promising results for purifying albumin to detect HD-adducts, exhibiting high purification efficiency, satisfactory albumin recovery, promising specificity, and a higher loading capacity for serum. PMID- 16267815 TI - Development of a simplified, economical polyacrylamide gel staining protocol for phosphoproteins. AB - Pro-Q Diamond (Pro-Q DPS) is a commercially available stain that binds the phosphate moiety of phosphoproteins with high sensitivity and linearity. To conserve consumable costs we demonstrate that threefold diluted Pro-Q DPS offers the same sensitivity and linearity of signal to that obtained with undiluted Pro Q DPS. The optimal conditions for Pro-Q DPS indicate that fixation, staining, and destaining of gels longer than 1 h, 2 h, and four 30-min incubations, respectively, are not required. The fixation and destaining solutions, but not the threefold diluted Pro-Q DPS, can be re-used without compromising the signal intensity or linear dynamic range. This modified protocol of Pro-Q DPS reduces the cost at least by fourfold, making the stain economically attractive for large scale analysis of phosphoproteins. PMID- 16267816 TI - A quantitative proteomic analysis of growth factor-induced compositional changes in lipid rafts of human smooth muscle cells. AB - Signals that promote proliferation and migration of smooth muscle cells (SMC) have been implicated in pathologic growth of hollow organs. Members of the platelet-derived growth factor (PDGF) family, potent mitogens and motility factors for SMC, have been shown to signal through cholesterol-enriched lipid rafts. We recently demonstrated that PDGF-stimulated DNA synthesis in urinary tract SMC was dependent on the integrity of lipid rafts. Despite its known ability to rapidly alter discrete proteins within rafts, the effect of PDGF on overall raft protein composition is unknown. In this study, we employed isotope coded affinity tag (ICAT) analysis to evaluate PDGF-induced protein changes in lipid rafts of primary culture human SMC. Following acute (i.e., 15 min) exposure of SMC to PDGF, 23 proteins increased in rafts >20%. In contrast, raft localization of only three proteins increased after 12 h of PDGF treatment. Among the proteins that increased at 15 min were the glycophosphatidylinositol-anchored proteins Thy-1, 5'-nucleotidase, and CD55, the cytoskeletal proteins actin, actinin, tropomyosin-3 and -4, and the endocytosis-related proteins clathrin and beta-adaptin. In addition, eight Rho family members were localized to rafts by ICAT analysis. Collectively, these observations suggest a role for lipid rafts in regulation of PDGF-stimulated changes in the cytoskeleton. PMID- 16267817 TI - Target identification of the novel antiobesity agent tungstate in adipose tissue from obese rats. AB - Adipose tissue plays an active role in the development of obesity, and thus characterization of the molecular changes related to obesity in this tissue is a priority. Recently, we identified tungstate as a potent body weight reducing agent in obese animals, adipose tissue being one of the targets of its action. In this study a proteomics approach combining 2-DE and MS was used to identify proteins associated with obesity and targets of tungstate in white adipose tissue. Twenty-nine proteins were found differentially expressed between lean and diet-induced obese rats. Expression changes in transferrin, vimentin, vinculin, peroxiredoxins, Rho-GTP dissociation inhibitor, grifin, guanine deaminase and 3 phosphoglycerate dehydrogenase were associated here for the first time with obesity. Furthermore, tungstate treatment of obese rats reverted expression changes of 70% of the proteins modulated by obesity and another ten proteins were regulated by tungstate independently of the body weight reduction. The results suggest that the tungstate antiobesity effect can be mediated by the modulation of cellular structure, metabolism, redox state and signalling processes in adipose tissue. These findings open new avenues for the study of the aetiology of obesity and its treatment. PMID- 16267819 TI - Fluoroprofile, a fluorescence-based assay for rapid and sensitive quantitation of proteins in solution. AB - The development of a sensitive fluorescence-based assay for the quantitative determination of protein concentration is described. The assay is based on the natural product epicocconone, which produces a large increase in fluorescence quantum yield upon binding to detergent-coated proteins in solution. There is a concomitant shift in the emission maximum from 520 to 605 nm after binding, which results in low background signal allowing a linear dynamic range of 40 ng/mL to 200 microg/mL for most proteins. There is little protein-to-protein variation except for iron-containing proteins and the assay can be used so that it is tolerant of chemicals commonly used in 2-D sample buffers. The assay is more sensitive than standard absorption assays such as the Bradford and Lowry assays, and has a greater dynamic range and sensitivity than other fluorescent assays. PMID- 16267818 TI - A proteomics strategy for the enrichment of receptor-associated complexes. AB - Multimeric protein complexes are important for cell function and are being identified by proteomics approaches. Enrichment strategies, such as those employing affinity matrices, are required for the characterization of such complexes, for example, those containing growth factor receptors. The receptor for the macrophage lineage growth factor, macrophage-colony stimulating factor (M CSF or CSF-1), is the tyrosine kinase, c-Fms. There is evidence that the CSF-1 receptor (CSF-1R) forms distinct multimeric complexes involving autophosphorylated tyrosines in its cytoplasmic region; however, these complexes are difficult to identify by immunoprecipitation, making enrichment necessary. We report here the use of a tyrosine-phosphorylated, GST-fusion construct of the entire CSF-1R cytoplasmic region to characterize proteins putatively associating with the activated CSF-1R. Besides signalling molecules known to associate with the receptor or be involved in CSF-1R-dependent signalling, mass spectrometry identified a number of other molecules binding to the construct. So far among these candidate proteins, dynein, claudin and silencer of death domains co immunoprecipitated with the CSF-1R, suggesting association. This affinity matrix method, using an entire cytoplasmic region, may have relevance for other growth factor receptors. PMID- 16267820 TI - Proteome analysis of a human uveal melanoma primary cell culture by 2-DE and MS. AB - We present here the first proteomics analysis of uveal melanoma (UM) cells. These cells represent a good model for the identification of polypeptide markers, which could be developed as diagnostic tools. UM is the most common primary intraocular tumour in adults. In contrast to other cancers, the survival rate of patients with this malignancy has changed little over the past few decades; a better understanding of the molecular biology of UM oncogenesis and metastasis is needed to build the basis for the identification of novel drug targets. In the study presented here, proteins from a UM primary cell culture were separated by 2-DE using a pI 3-10 gradient; 270 spots were analysed by LC-MS/MS, identifying 683 proteins derived from 393 different genes. Of those, 69 (18%) are related to cancer processes involving cell division, proliferation, invasion, metastasis, oncogenesis, drug resistance and others. To our knowledge, 96% of the proteins identified, including 16 hypothetical proteins, have never been reported in UM before. This study represents the first step towards the establishment of a UM protein database as a valuable resource for the study of this malignancy. PMID- 16267822 TI - Live imaging and morphometric analysis of embryonic development in the ascidian Ciona intestinalis. AB - The ascidian Ciona intestinalis is one of the model organisms of choice for comparative investigations of chordate development and for unraveling the molecular mechanisms underlying morphogenesis and cell fate specification. Taking advantage of the availability of various genetically encoded fluorescent proteins and of defined cis-regulatory elements, we combined transient transgenesis with laser scanning confocal imaging to acquire and quantitate 3D time-lapse data from living Ciona embryos. We used Ciona tissue-specific enhancers to drive expression of spectrally distinct fluorescent protein reporters to label and simultaneously visualize axially and paraxially positioned mesodermal derivatives, as well as neural precursors in individual embryos. We observed morphogenetic movements, without perturbing development, from the early gastrula throughout the larval stage, including gastrulation, neurulation, convergent extension of the presumptive notochord, and tail elongation. These multidimensional data allowed us to establish a reference system of metrics to quantify key developmental events including blastopore closure and muscle extension. The approach we describe can be used to document morphogenetic cell and tissue rearrangements in living embryos and paves the way for a live digitized anatomical atlas of Ciona. PMID- 16267821 TI - In vivo genetic ablation by Cre-mediated expression of diphtheria toxin fragment A. AB - We generated a ROSA26-eGFP-DTA mouse line by introducing an eGFP-DTA (enhanced green fluorescent protein -- diphtheria toxin fragment A) cassette into the ROSA26 locus by homologous recombination in ES cells. This mouse expresses eGFP ubiquitously, but DTA expression is prevented by the presence of eGFP, a Neo cassette, and a strong transcriptional stop sequence. Mice carrying this construct are normal and fertile, indicating the absence of DTA expression. However, upon Cre-mediated excision of the floxed region DTA expression is activated, resulting in the specific ablation of Cre-expressing cells. As an example of this approach, we ablated Nkx2.5 and Wnt1-expressing cells by using the Nkx2.5-Cre and Wnt1-Cre mouse lines, respectively. We observed loss of the precise tissues in which Nkx2.5 and Wnt1 are expressed. Apart from being a general GFP reporter, the ROSA26-GFP-DTA mouse line should provide a useful resource for genetic ablation of specific groups of cells. PMID- 16267824 TI - Peroxisome proliferator-activated receptors: potential therapeutic targets in lung disease? AB - The peroxisome proliferator-activated receptors (PPARs) are a family of nuclear hormone receptors that play central roles in lipid and glucose homeostasis, cellular differentiation, and the immune/inflammatory response. Growing evidence indicates that changes in expression and activation of PPARs likely modulate conditions as diverse as diabetes, atherosclerosis, cancer, asthma, Parkinson's disease, and Alzheimer's disease. Activation of these receptors by natural or pharmacologic ligands leads to both gene-dependent and gene-independent effects that alter the expression of a wide array of proteins. In the lung, PPARs are expressed by alveolar macrophages, as well as by epithelial, endothelial, and smooth muscle cells. Studies both in vitro and in vivo suggest that PPAR ligands may have anti-inflammatory effects in asthma, pulmonary sarcoidosis, and pulmonary alveolar proteinosis, as well as antiproliferative and antiangiogenic effects in epithelial lung cancers. Further studies to understand the contribution of these receptors to health and disease will be important for determining whether they represent a promising target for therapeutic intervention. PMID- 16267823 TI - Meox1Cre: a mouse line expressing Cre recombinase in somitic mesoderm. AB - We developed a novel strategy based on in vitro DNA transposition of phage Mu to construct vectors for "knock-in" of the gene encoding Cre recombinase into endogenous loci in embryonic stem cells. This strategy was used to introduce Cre into the mouse Meox1 locus, which was expected to drive Cre expression in the presomitic and somitic mesoderm. In embryos heterozygous for both Meox1(Cre) and R26R or Z/AP reporter alleles, specific and efficient recombination of the reporter alleles was detected in the maturing somites and their derivatives, including developing vertebrae, skeletal muscle, back dermis, as well as endothelium of the blood vessels invading the spinal cord and developing limbs. In contrast to the somitic mesoderm, Cre activity was not observed in the cranial paraxial mesoderm. Thus, the Meox1(Cre) allele allows detailed fate-mapping of Meox1-expressing tissues, including derivatives of the somitic mesoderm. We used it to demonstrate dynamic changes in the composition of the mesenchyme surrounding the developing inner ear. Meox1(Cre) may also be used for tissue specific mutagenesis in the somitic mesoderm and its derivatives. PMID- 16267826 TI - The facial motor nucleus transcriptional program in response to peripheral nerve injury identifies Hn1 as a regeneration-associated gene. AB - Facial nerve axotomy (FNA) is a well-established experimental model of motoneuron regeneration. After peripheral nerve axotomy, a sequence of events including glial activation and axonal regrowth leads to functional recovery of the afflicted pool of motoneurons. Using microarray analysis we identified an increase in the expression of 60 genes (at a false discovery rate of 0.1, genes were significant P < 0.004) within the facial nucleus as a consequence of nerve injury. In situ hybridization analysis validated the increased expression of many of these axotomy-induced genes. One specific gene, encoding a unique primary amino acid sequence, termed hemopoietic- and neurologic-expressed sequence-1 (Hn1), was evaluated more extensively using several additional nerve injury paradigms. Hn1 mRNA was upregulated in injured facial motoneurons in both rats and mice. Sustained upregulation of Hn1 mRNA was evident after nerve resection whereas levels of Hn1 mRNA returned to baseline in animals subjected to nerve crush or nerve transection. Hn1 was also increased in the dorsal motor nucleus and the nucleus ambiguous after vagus nerve axotomy, another regeneration model. No upregulation of Hn1 expression was observed, however, in two nonregeneration models: FNA in newborn rats and rubrospinal tractotomy. Hn1 mRNA was ubiquitous in the developing central nervous system whereas its expression in adult brain was confined to neurons of the hippocampus, cortex and cerebellum. These findings identify Hn1 as a gene associated with nervous system development and nerve regeneration. PMID- 16267825 TI - Neuroprotective effects of GluR6 antisense oligodeoxynucleotides on transient brain ischemia/reperfusion-induced neuronal death in rat hippocampal CA1 region. AB - To investigate whether the kainate (KA) receptors subunit GluR6 is involved in the neuronal cell death induced by cerebral ischemia followed by reperfusion, the antisense oligodeoxynucleotides (ODNs) of GluR6 were used to suppress the expression of GluR6 by intracerebroventricular infusion once per day for 3 days before ischemia. Transient brain ischemia was induced by four-vessel occlusion in Sprague-Dawley rats. The effects of GluR6 antisense ODNs on the phosphorylation of MLK3 and JNK and the interactions of MLK3 and PSD-95 with GluR6 were examined by immunoprecipitation and immunoblotting. Our results show that GluR6 antisense ODNs can knock down the expression of GluR6 and suppress the assembly of the GluR6.PSD-95.MLK3 signaling module and, therefore, inhibit JNK activation and phosphoralation of c-jun. On the other hand, the GluR6 antisense ODNs also show a protective role against neuronal cell death induced by cerebral ischemia/reperfusion. Administration of GluR6 antisense ODNs once per day for 3 days before cerebral ischemia significantly decreased neuronal degeneration. In conclusion, our results demonstrate that kainate receptor subunit GluR6 plays an important role in neuronal death induced by cerebral ischemia followed by reperfusion. PMID- 16267827 TI - Differential role of tumor necrosis factor receptors in mouse brain inflammatory responses in cryolesion brain injury. AB - Tumor necrosis factor-alpha (TNF-alpha) is one of the mediators dramatically increased after traumatic brain injury that leads to the activation, proliferation, and hypertrophy of mononuclear, phagocytic cells and gliosis. Eventually, TNF-alpha can induce both apoptosis and necrosis via intracellular signaling. This cytokine exerts its functions via interaction with two receptors: type-1 receptor (TNFR1) and type-2 receptor (TNFR2). In this work, the inflammatory response after a freeze injury (cryolesion) in the cortex was studied in wild-type (WT) animals and in mice lacking TNFR1 (TNFR1 KO) or TNFR2 (TNFR2 KO). Lack of TNFR1, but not of TNFR2, significantly decreased the inflammatory response and tissue damage elicited by the cryolesion at both 3 and 7 days postlesion, with decreased gliosis, lower IL-1beta immunostaining, and a reduction of apoptosis markers. Cryolesion produced a clear induction of the proinflammatory cytokines interleukin (IL)-1alpha, IL-1beta, IL-6, and TNF-alpha; this induction was significantly lower in the TNFR1 KO mice. Host response genes (ICAM-1, A20, EB22/5, and GFAP) were also induced by the cryolesion, but to a lesser extent in TNFR1 KO mice. Lack of TNFR1 signaling also affected the expression of apoptosis/cell death-related genes (Fas, Rip, p53), matrix metalloproteinases (MMP3, MMP9, MMP12), and their inhibitors (TIMP1), suggesting a role of TNFR1 in extracellular matrix remodeling after injury. However, GDNF, NGF, and BDNF expression were not affected by TNFR1 deficiency. Overall, these results suggest that TNFR1 is involved in the early establishment of the inflammatory response and that its deficiency causes a decreased inflammatory response and tissue damage following brain injury. PMID- 16267828 TI - Phosphodiesterase-Ialpha/autotaxin (PD-Ialpha/ATX): a multifunctional protein involved in central nervous system development and disease. AB - Phosphodiesterase-Ialpha/autotaxin (PD-Ialpha/ATX) was originally identified as a cell-motility-stimulating factor secreted by a variety of tumor cells. Thus, studies related to its potential functional roles have traditionally focused on tumorigenesis. PD-Ialpha/ATX's catalytic activity, initially defined as nucleotide pyrophosphatase/phosphodiesterase, was soon recognized as being necessary for its tumor cell-motility-stimulating activity. However, only the discovery of PD-Ialpha/ATX's identity with lysophospholipase D, an extracellular enzyme that converts lysophosphatidylcholine into lysophosphatidic acid (LPA) and potentially sphingosylphosphoryl choline into sphingosine 1-phosphate (S1P), revealed the actual effectors responsible for PD-Ialpha/ATX's ascribed motogenic functions, i.e., its catalytic products. PD-Ialpha/ATX has also been detected during normal development in a number of tissues, in particular, the central nervous system (CNS), where expression levels are high. Similar to tumor cells, PD-Ialpha/ATX-expressing CNS cells secrete catalytically active PD-Ialpha/ATX into the extracellular environment. Thus, it appears reasonable to assume that PD Ialpha/ATX's CNS-related functions are mediated via lysophospholipid, LPA and potentially S1P, signaling. However, recent studies identified PD-Ialpha/ATX as a matricellular protein involved in the modulation of oligodendrocyte-extracellular matrix interactions and oligodendrocyte remodeling. This property of PD Ialpha/ATX was found to be independent of its catalytic activity and to be mediated by a novel functionally active domain. These findings, therefore, uncover PD-Ialpha/ATX, at least in the CNS, as a multifunctional protein able to induce complex signaling cascades via distinct structure-function domains. This Mini-Review describes PD-Ialpha/ATX's multifunctional roles in the CNS and discusses their potential contributions to CNS development and pathology. PMID- 16267829 TI - The cytoplasmic tail of the alpha3 integrin subunit promotes neurite outgrowth in PC12 cells. AB - Binding of integrins to proteins of the extracellular matrix (ECM) provides structural and signaling information for biological processes such as cell proliferation, migration, neurite outgrowth, and differentiation. Integrins represent a family of heterodimeric transmembrane cell surface receptors. Besides connecting the ECM with the cytoskeleton, integrins also induce various signaling pathways in response to ligand binding. Integrin ligation leads to cytoplasmic protein-protein interactions requiring both integrin cytoplasmic tails. These sequences are initiation points for focal adhesion formation and subsequent signal transduction cascades. In this study, we addressed the question of whether the short cytoplasmic tail of the alpha(3) integrin subunit of alpha(3)beta(1) integrin is required for alpha(3)beta(1) integrin-dependent processes. For this purpose, cDNA representing the extracellular and transmembrane domain of the interleukin 2 receptor (IL2R) alpha subunit and the cytoplasmic sequence of the alpha(3) integrin subunit was transfected into PC12 cells. Autonomous expression of the cytoplasmic alpha(3) tail does not affect attachment but leads to inhibition of neuronal differentiation on laminin 5. This indicates that the cytoplasmic alpha(3) sequence is not required for cell attachment but is necessary for long-term adhesion and for the reorganization of the cytoskeleton that precedes neuronal differentiation. Inhibition of neurite outgrowth by chimeric IL2R-alpha(3) can be rescued by treatment of transfected cells with the pharmacological inhibitor Y27632, which inhibits the RhoA downstream effector Rho kinase alpha. PMID- 16267830 TI - Species-specific effects of the hepatocarcinogens 3'-methyl-4-dimethyl aminoazobenzene and ortho-aminoazotoluene in mouse and rat liver. AB - The effects of rat-specific hepatocarcinogen 3'-methyl-4-dimethylaminoazobenzene (3'-MeDAB), mouse-specific hepatocarcinogen ortho-aminoazotoluene (OAT), non species-specific hepatocarcinogen diethylnitrosamine (DENA), and non-carcinogenic 4'-methyl-4-dimethylaminoazobenzene (4'-MeDAB) on glucocorticoid induction of tyrosine aminotransferase (TAT) and DNA-binding activity of hepatocyte nuclear factor 3 (HNF3) family of transcription factors were investigated with carcinogen susceptible and -resistant animals. Species-specific hepatocarcinogens 3'-MeDAB and OAT strongly inhibited glucocorticoid induction of TAT in the liver of susceptible but not resistant animals. DENA, which is highly carcinogenic for the liver of both rats and mice inhibited glucocorticoid induction of TAT in both species, while non-carcinogenic 4'-MeDAB was absolutely ineffective both in rats and mice. The inhibition of TAT activity by the carcinogens was due to reduced levels of TAT mRNA, which is most likely to be a result of the reduced rate of transcription initiation of the TAT gene. In all cases, the TAT inhibition was accompanied by significant reduction of DNA-binding activity of the HNF3 transcription factor, which is known to be critical to glucocorticoid regulation of TAT gene. We also demonstrated that the described species-specific effects of OAT and of 3'-MeDAB on HNF3 DNA-binding activity may be initiated not only by administration in vivo, but also by their direct administration to homogenate, intact nuclei or nuclear lysate, but not to nuclear extract fraction, obtained by precipitation with 0.32 g/mL of ammonium sulfate (Fraction I). We showed, that a factor responsible for this effect might be precipitated in 0.32-0.47 g/mL interval of ammonium sulfate concentration. In contrast, non-specific hepatocarcinogen DENA was effective upon being added directly to Fraction I, implying a different mechanism of its action. PMID- 16267831 TI - Mutant human tumor suppressor p53 modulates the activation of mitogen-activated protein kinase and nuclear factor-kappaB, but not c-Jun N-terminal kinase and activated protein-1. AB - The roles of the mitogen-activated kinase protein (MAPK) pathway, nuclear factor kappa B (NF-kappaB), and activator protein-1 (AP-1) in cellular responses to growth factors and mitogen are well established. However, the manner by which these proliferative pathways are affected by the tumor suppressor protein p53 is not fully understood. We report here the results of an investigation of the status of p53 on two human melanoma cell lines with wild-type p53 (SK-Mel-186) or mutant p53 (SK-Mel-110). The basal levels of the activated extracellular-signal regulated kinases 1 and 2 (ERK1/2) were high in cells with wild-type p53, but low in cells with mutant p53. The 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced activation of ERK1/2 through the phosphorylation of threonine and tyrosine at 202 and 204, respectively, was demonstrated in both cell lines, however, in a discrete manner. TPA-induced activation of ERK1/2 was sustained in wild-type p53 cells, while only a transient activation was seen in mutant p53 cells. Inhibition of MAPK kinase (MEK), an upstream kinase, by U0126, blocked TPA-induced activation of ERK1/2 in wild-type p53 cells and in mutant p53 cells. Treatment of wild-type p53 (SK-Mel 186) cells with small interfering RNA (siRNA) of p53 displayed a transient induction of activation of ERK1/2 following TPA treatment, indicating that p53 has a role in the regulation of the activation of ERK1/2. NF kappaB activity decreased significantly in cells with wild-type p53, while enhanced NF-kappaB activity was evident in cells with mutant p53. The expression of either wild-type or mutant p53 had a similar effect on TPA-induced Jun N terminal kinase (JNK) activation, indicating specificity for the ERK pathway. Similarly, AP-1 binding activity showed a transient variation in both cell lines after TPA treatment but with different kinetics. These observations suggest that both wild-type and mutant p53 can modulate the activation pathways for ERK1/2, and NF-kappaB distinctively, while modulating the pathways of JNK and AP-1 similarly. These differences may influence cellular processes such as proliferation, differentiation, and apoptosis. PMID- 16267832 TI - Synergistic interaction between 17-AAG and phosphatidylinositol 3-kinase inhibition in human malignant glioma cells. AB - The phosphatidylinositol 3'-kinase (PI3K)/Akt pathway is often constitutively activated in malignant glioma cells, in many cases as a result of mutation of phosphatase and tensin homologue deleted on chromosome ten (PTEN), an endogenous inhibitor of Akt, which renders tumor cells resistant to cytotoxic insults, including those related to anticancer drugs. Pharmacological inhibition of this pathway may potentially restore or augment the effectiveness of conventional chemotherapy or other signaling-targeted agents. Because the heat shock protein (HSP) is involved in the conformational maturation of a number of signaling proteins critical to the proliferation of malignant glioma cells, we hypothesized that the combination of the PI3K inhibitor LY294002 and the HSP90 inhibitor 17 allyl-aminogeldanamycin (17-AAG) would promote glioma cytotoxicity by decreasing both the activation status and levels of Akt, as well as downregulating the levels of other relevant signaling effectors. We, therefore, examined the effects of LY294002 and 17-AAG, alone and in combination, on signal transduction and apoptosis in a series of malignant glioma cell lines. Simultaneous exposure to these inhibitors significantly induced cell death, and irreversibly inhibited proliferative activity and colony forming ability of the glioma cell lines. Quantitative analysis revealed that enhancement by LY294002 of 17-AAG-induced cytotoxicity was synergistic, leading to a pronounced increase in active caspase 3 and poly (adenosine diphosphate-ribose) polymerase (PARP) cleavage together with the release of cytochrome c and apoptosis inducing factor (AIF). No significant growth inhibition or caspase activation was seen in control cells. The enhanced cytotoxicity of this combination was associated with diminished Akt activation and a significant downregulation of epidermal growth factor receptor (EGFR), Raf-1, and mitogen activated protein kinase. Combination of 17-AAG and LY294002 did not modify phospho-JNK/SPK and phospho-p38. Cells exposed to 17-AAG and LY294002 displayed a significant reduction in cell-cycle regulatory proteins, such as retinoblastoma (Rb), cyclin dependent kinase (CDK)4, CDK6, cyclin D1, and cyclin D3. Taken together, these findings suggest that the PI3K/Akt pathway plays a critical role in regulating the apoptotic response to 17-AAG and that targeting this pathway could provide a potent strategy to treat patients with malignant gliomas. PMID- 16267833 TI - Labeled Schwann cell transplantation: cell loss, host Schwann cell replacement, and strategies to enhance survival. AB - Although transplanted Schwann cells (SCs) can promote axon regeneration and remyelination and improve recovery in models of spinal cord injury, little is known about their survival and how they interact with host tissue. Using labeled SCs from transgenic rats expressing human placental alkaline phosphatase (PLAP), SC survival in a spinal cord contusion lesion was assessed. Few PLAP SCs survived at 2 weeks after acute transplantation. They died early due to necrosis and apoptosis. Delaying transplantation until 7 days after injury improved survival. A second wave of cell death occurred after surviving cells had integrated into the spinal cord. Survival of PLAP SCs was enhanced by immunosuppression with cyclosporin; delayed transplantation in conjunction with immunosuppression resulted in the best survival. In all cases, transplantation of SCs resulted in extensive infiltration of endogenous p75+ cells into the injury site, suggesting that endogenous SCs may play an important role in the repair observed after SC transplantation. PMID- 16267834 TI - Phenotypic and functional heterogeneity of GFAP-expressing cells in vitro: differential expression of LeX/CD15 by GFAP-expressing multipotent neural stem cells and non-neurogenic astrocytes. AB - Recent findings show that the predominant multipotent neural stem cells (NSCs) isolated from postnatal and adult mouse brain express glial fibrillary acid protein (GFAP), a protein commonly associated with astrocytes, and that primary astrocyte cultures can contain GFAP-expressing cells that act as multipotent NSCs when transferred to neurogenic conditions. The relationship of GFAP-expressing NSCs to GFAP-expressing astrocytes is unclear, but has important implications. We compared the phenotype and neurogenic potential of GFAP-expressing cells derived from different CNS regions and maintained in vitro under different conditions. Multiple labeling immunohistochemistry revealed that both primary astrocyte cultures and adherent neurogenic cultures derived from postnatal or adult periventricular tissue contained subpopulations of GFAP-expressing cells that co expressed nestin and LeX/CD15, two molecules associated with NSCs. In contrast, GFAP-expressing cells in similar cultures prepared from adult cerebral cortex did not express detectable levels of LeX/CD15, and exhibited no neurogenic potential. Fluorescence-activated cell sorting (FACS) of both primary astrocyte cultures and adherent neurogenic cultures for LeX/CD15 showed that GFAP-expressing cells competent to act as multipotent NSCs were concentrated in the LeX-positive fraction. Using neurosphere assays and a transgenic ablation strategy, we confirmed that the predominant NSCs in primary astrocyte and adherent neurogenic cultures were GFAP-expressing cells. These findings demonstrate that GFAP expressing cells derived from postnatal and adult forebrain are heterogeneous in both molecular phenotype and neurogenic potential in vitro, and that this heterogeneity exists before exposure to neurogenic conditions. The findings provide evidence that GFAP-expressing NSCs are phenotypically and functionally distinct from non-neurogenic astrocytes. PMID- 16267836 TI - Prevalent mutations in prostate cancer. AB - Quantitative and structural genetic alterations cause the development and progression of prostate cancer. A number of genes have been implicated in prostate cancer by genetic alterations and functional consequences of the genetic alterations. These include the ELAC2 (HPC2), MSR1, and RNASEL (HPC1) genes that have germline mutations in familial prostate cancer; AR, ATBF1, EPHB2 (ERK), KLF6, mitochondria DNA, p53, PTEN, and RAS that have somatic mutations in sporadic prostate cancer; AR, BRCA1, BRCA2, CHEK2 (RAD53), CYP17, CYP1B1, CYP3A4, GSTM1, GSTP1, GSTT1, PON1, SRD5A2, and VDR that have germline genetic variants associated with either hereditary and/or sporadic prostate cancer; and ANXA7 (ANX7), KLF5, NKX3-1 (NKX3.1), CDKN1B (p27), and MYC that have genomic copy number changes affecting gene function. More genes relevant to prostate cancer remain to be identified in each of these gene groups. For the genes that have been identified, most need additional genetic, functional, and/or biochemical examination. Identification and characterization of these genes will be a key step for improving the detection and treatment of prostate cancer. PMID- 16267835 TI - Lymphocyte enhancer-binding factor 1 (Lef1) inhibits terminal differentiation of osteoblasts. AB - Lef1 is a transcriptional regulator of the Wnt/beta-catenin signaling cascade. Wnts directly augment bone formation and osteoblast differentiation from mesenchymal stem cells by receptor-mediated pathways involving Lrp5 and Frizzled. We previously reported that Lef1 represses Runx2-dependent activation of the late osteoblast differentiation gene, osteocalcin. Lef1 is expressed in preosteoblasts but is undetectable in fully differentiated osteoblasts. To determine if downregulation of Lef1 is necessary for osteoblast maturation, we constitutively overexpressed Lef1 in MC3T3-E1 preosteoblasts. Lef1-overexpressing cells produced alkaline phosphatase (ALP) and osteocalcin later, and at lower levels than control cells. Moreover, the extracellular matrices of Lef1-overexpressing cell cultures never mineralized. To further examine the role of Lef1 in osteoblasts, we suppressed Lef1 expression in MC3T3-E1 cells by RNA interference. Transient expression of a Lef1 shRNA efficiently reduced murine Lef1 levels and transcriptional activity. Stable suppression of Lef1 in MC3T3 preosteoblasts did not affect proliferation or Runx2 levels; however, ALP production and matrix mineralization were accelerated by 3-4 days. Gene chip analyses identified 14 genes that are differentially regulated in Lef1-suppressed cells. These data outline a role for Lef1 in delaying osteoblast maturation and suggest that Lef1 controls the expression of multiple genes in osteoblasts. PMID- 16267837 TI - Cell cycle control in breast cancer cells. AB - In breast cancer, cyclins D1 and E and the cyclin-dependent kinase inhibitors p21 (Waf1/Cip1)and p27 (Kip1) are important in cell-cycle control and as potential oncogenes or tumor suppressor genes. They are regulated in breast cancer cells following mitogenic stimuli including activation of receptor tyrosine kinases and steroid hormone receptors, and their deregulation frequently impacts on breast cancer outcome, including response to therapy. The cyclin-dependent kinase inhibitor p16 (INK4A) also has a critical role in transformation of mammary epithelial cells. In addition to their roles in cell cycle control, some of these molecules, particularly cyclin D1, have actions that are not mediated through regulation of cyclin-dependent kinase activity but may be important for loss of proliferative control during mammary oncogenesis. PMID- 16267838 TI - An osteoclastic protein-tyrosine phosphatase is a potential positive regulator of the c-Src protein-tyrosine kinase activity: a mediator of osteoclast activity. AB - This study tested the hypothesis that an osteoclastic protein-tyrosine phosphatase, PTP-oc, enhances osteoclast activity through c-Src activation. The effects of several resorption activators and inhibitors on PTP-oc expression, resorption activity, and c-Src activation were determined in rabbit osteoclasts. PTP-oc expression was assayed with immunoblots and semi-quantitative RT-PCR. Osteoclastic activity was determined by the resorption pit assay; and c-Src activation was monitored by P-tyr527 (PY527) dephosphorylation, and in vitro kinase assay. Treatment of osteoclasts with PTH, PGE2, 1,25(OH)2D3, IL-1, but not RANKL or IL-6, significantly stimulated resorption activity, increased PTP-oc mRNA and protein levels, and reduced c-Src PY527 level with corresponding activation of c-Src protein-tyrosine kinase activity. The PTP-oc antisense phosphorothioated oligo treatment blocked the basal and IL-1alpha-mediated, but not RANKL-mediated, resorption activity of isolated osteoclasts. The antisense oligo treatment also significantly reduced the average depth of resorption pits created by rabbit osteoclasts under basal conditions. Calcitonin and alendondrate, significantly reduced resorption activity and PTP-oc expression, and increased c-Src PY527 with corresponding reduction in its PTK activity. The cellular PTP-oc protein level correlated with the resorption activity. Among the various signaling proteins co-immunoprecipitated with PTP-oc, the resorption effectors caused corresponding changes in the tyrosyl phosphorylation level of only c-Src. The GST-PTP-oc fusion protein dephosphorylated PY-527-containing c Src peptide in time- and dose-dependent manner in vitro. In summary, (1) PTP-oc is regulated in part at transcriptional level, (2) upregulation of PTP-oc in osteoclasts led to c-Src activation, and (3) PY527 of c-Src may be a cellular substrate of PTP-oc. These findings are consistent with the hypothesis that PTP oc is a positive regulator of c-Src in osteoclasts. PMID- 16267839 TI - Nuclear phosphoinositide kinases and inositol phospholipids. AB - The presence of inositol phospholipids in the nuclei of mammalian cells has by now been well established, as has the presence of the enzymes responsible for their metabolism. However, our understanding of the role of these nuclear phosphoinositides in regulating cellular events has lagged far behind that for its cytosolic counterpart. It is clear, though, that the nuclear phosphoinositide pool is independent of the cytosolic pool and is, therefore, likely to be regulating a unique set of cellular events. As with its cytosolic phosphoinositides, many nuclear phosphoinositides and their metabolic enzymes are located at distinct sub-cellular structures. This arrangement spatially limits the production and activity of inositol phospholipids and is believed to be a major mechanism for regulating their function. Here, we will introduce the components of nuclear inositol phospholipid signal transduction and discuss how their spatial arrangement may dictate which nuclear functions they are modulating. PMID- 16267840 TI - Albumin induces cellular fibrosis by upregulating transforming growth factor-beta ligand and its receptors in renal distal tubule cells. AB - Albuminuria is indicative of nephropathy. However, little literature has focused on the role of albumin in renal distal tubule fibrosis. We used a well-defined distal tubule cell, Madin-Darby Canine Kidney (MDCK). Proliferation and cytotoxicity were examined. The conditioned supernatant was collected and subjected to ELISA assay for detection of fibronectin and TGF-beta1. Reverse transcription-PCR and Western blot assay were performed to evaluate the expression of mRNA and protein of two types of TGF-beta receptors (TbetaR). Flow cytometry assay and phosphotyrosine (pY)-specific antibodies were used to assay the phosphorylation status of TbetaR. We showed that albumin dose dependently (0, 0.1, 1, or 10 mg/ml) inhibited cellular growth in MDCK cells without inducing cellular cytotoxicity. In addition, albumin significantly upregulated the secretion of both fibronectin and TGF-beta1 at dose over 1 mg/ml. Moreover, 24 h pretreatment of albumin significantly enhanced exogenous TGF-beta1-induced secretion of fibronectin. These observations were reminiscent of the implications of TbetaR since TbetaR appears to correlate with the susceptibility of cellular fibrosis. We found that albumin significantly increased protein levels of type I TbetaR (TbetaRI) instead of type II receptors (TbetaRII). In addition, phosphorylation level of TbetaRII of both pY259 and pY424 was significantly enhanced instead of pY336. The novel observation indicates that extreme dose of albumin upregulates TGF-beta autocrine loop by upregulating TGF-beta1, TbetaRI, and the receptor kinase activity of TbetaRII by inducing tyrosine phosphorylation on key amino residue of TbetaRII in renal distal tubule cells. These combinational effects might contribute to the pathogenesis of renal fibrosis. PMID- 16267841 TI - Identification of estrogen-responsive genes in the testis of sea bream (Sparus auratus) using suppression subtractive hybridization. AB - There is growing evidence that estrogens play important roles in both normal and xenoestrogen disrupted testis physiology. However, the mechanisms and signaling pathways involved, in particular in fish, are largely unknown. We have used suppression subtractive hybridization to isolate 152 candidate estrogen responsive genes in the testis of male estradiol (E2)-treated sea bream (Sparus aurata). The E2 up-regulation of some of the genes (e.g., choriogenin L and H, vitellogenin I and II, apolipoprotein A-I, fibrinogen beta and gamma, and thyroid receptor interacting protein 4) was confirmed by reverse transcriptase polymerase chain reaction in fish treated with 0.1-10 mg/kg E2. Many of these genes are typical E2-induced genes in liver, and this is the first report of its up regulation with E2 in testis. Moreover, low levels of expression were also found for nontreated fish. Hepatic differential expression for these genes was also confirmed, although, contrary to testis, fibrinogen beta, and gamma were downregulated. The possible significance of these findings in normal testis physiology and in endocrine disruption is discussed. PMID- 16267843 TI - Neurocysticercosis-induced hemichorea. PMID- 16267842 TI - The Parkinson-Control study: a 1-year randomized, double-blind trial comparing piribedil (150 mg/day) with bromocriptine (25 mg/day) in early combination with levodopa in Parkinson's disease. AB - Dopamine agonists have been recommended as early treatment for Parkinson's disease (PD), alone or combined with levodopa. Piribedil is a non-ergot selective D(2)/D(3) agonist with alpha(2) antagonist properties shown to be effective in the treatment of PD. This 12-month international, randomized, double-blind trial aimed to assess the efficacy of piribedil 150 mg versus bromocriptine 25 mg, in early combination with levodopa in Stage I to III PD patients. Motor efficacy was assessed using the Unified Parkinson's Disease Rating Scale (UPDRS III, Items 18 31) as improvement from baseline. Response rate was defined as a 30% improvement. Among the 425 randomly assigned patients, 178 were also included in a substudy on cognitive follow-up evaluated by a dysexecutive syndrome oriented battery. A relevant improvement in UPDRS III over the 12-month study duration was observed both in the piribedil and bromocriptine groups (-7.9 +/- 9.7 points from baseline versus -8.0 +/- 9.5; not significant [n.s.]) with a response rate of 58.4% and 55.3% (n.s.), respectively. Piribedil and bromocriptine resulted in similar improvement on all UPDRS III subscores. Piribedil patients required less levodopa dose increase than those on bromocriptine. Cognitive performance remained generally unchanged in both groups, with a significant effect of piribedil limited to the Wisconsin Card Sorting Test. An overall good tolerability of piribedil was observed. Early combination of piribedil 150 mg with levodopa resulted in significant long-term improvement of all motor symptoms in PD patients insufficiently controlled by levodopa alone. Taking into account both efficacy and acceptability in the long-term, piribedil proved in this bromocriptine controlled study to be an effective and safe treatment for PD. PMID- 16267844 TI - Complete suppression of paroxysmal nonkinesigenic dyskinesia by globus pallidus internus pallidal stimulation. AB - Stereotactic functional surgery is being explored as potential therapies for medically intractable paroxysmal dyskinesias (PxD). We report on a 59-year-old man in whom stimulation of globus pallidus internus produced immediate and sustained relief of paroxysmal non-kinesigenic dyskinesia secondary to a rotator cuff tears on the left shoulder. Our finding strongly suggests that altered function of neuronal circuits of the basal ganglia underlies the manifestation of PxD. PMID- 16267845 TI - Dopa-responsive dystonia and early-onset Parkinson's disease in a patient with GTP cyclohydrolase I deficiency? AB - We describe a patient with a combination of dystonic and parkinsonian signs. Paraclinical studies revealed a mutation in the GTP cyclohydrolase I gene (GCH1) and a decrease in [123I]-N-omega-fluoropropyl-2beta-carbomethoxy-3beta-(4 iodophenyl) nortropane (123I-FP-CIT) binding ratios indicative of Parkinson's disease. We conclude that the patient probably suffers from a variant of dopa responsive dystonia (DRD) or two separate movement disorders, normally considered to be differential diagnoses, DRD and early-onset Parkinson's disease with resulting difficulties concerning treatment and prognosis. PMID- 16267847 TI - Genetic, clinical, and imaging characterization of one patient with late-onset, slowly progressive, pantothenate kinase-associated neurodegeneration. AB - We report on a patient with late-onset, pantothenate kinase-associated neurodegeneration (PKAN) who revealed two new heterozygous mutations at gene testing and showed asymmetric moderately reduced striatal dopamine transporter binding with single photon emission computed tomography, possibly due to prolonged neuroleptic treatment. These findings expand the genetic and imaging spectrum of this rare disorder. PMID- 16267846 TI - Clinical and genetic study of a Brazilian family with spastic paraplegia (SPG6 locus). AB - We describe a Brazilian family in which inheritance of a G106R mutation in the SPG6 gene (also know as NIPA1) resulted in an autosomal dominant form of hereditary spastic paraplegia (ADHSP). Clinical investigations indicated that this family has a pure form of spastic paraplegia. All patients presented with gait difficulty in their twenties, progressing to frank spastic paraplegia during the next decade. Our report further supports evidence that mutations in SPG6 cause ADHSP. PMID- 16267848 TI - Enhanced growth in NS0 cells expressing aminoglycoside phosphotransferase is associated with changes in metabolism, productivity, and apoptosis. AB - Prior work has reported that cotransfecting a gene of interest with the selectable marker neo can seriously perturb a number of cellular processes. In this study the influence of the neo gene on the growth, death, and metabolism of a murine myeloma NS0 cell line, expressing a chimeric antibody, was investigated. A pool of neo transfectants, 6A1-NEO, was selected with 500 microg/mL G418. Quantitative PCR analysis revealed that 6A1-NEO contained, on average, three copies of the neo gene per cell. Batch cultivation of 6A1-NEO showed that there was a 36% increase in maximum viable cell concentration, a 20% increase in the maximum apparent growth rate, and a 134% increase in cumulative cell hours as compared with the parent, 6A1-(100)3. Batch cultivation of five randomly selected clones illustrated that 6A1-NEO's advantage over the parent was not due to clonal variation. Neither the use of G418 during the selection process nor the cultivation of cells in the presence of G418 were responsible. This implied that the neo gene product, APH(3')-II, was causing the changes in proliferative capacity. Analysis of the cell cycle revealed that there were no differences in the distribution of cells in the G(1), S, and G(2) phases. When cell growth was synchronized, there were no observed differences in cell-cycle duration. 6A1-NEO resisted the onset of apoptosis during the growth phase. Consequently, there was a larger viable population of 6A1-NEO cells available for proliferation as compared with the parent. However, 6A1-NEO died at the same rate as the parent when resuspended in spent media or after treatment with staurosporin. Expression of the anti-apoptotic protein Bcl-2 was upregulated in 6A1-NEO, indicating that APH(3')-II could be acting by modulating endogenous gene expression. Analysis of key metabolites showed that 6A1-NEO's specific glucose consumption rate was 133% higher, whilst its specific glutamate consumption rate was 45% lower than the parent. 6A1-NEO's efficient utilization of glutamate and shift towards glucose metabolism may have contributed to the rise in proliferative capacity. However, this was accompanied by a 70% drop in the specific antibody production rate. These results show that the increase in growth rate and proliferative capacity caused by the expression of recombinant APH(3')-II was associated with changes in metabolism, apoptosis, and endogenous gene expression. PMID- 16267849 TI - Process modeling of in situ-adsorption of a bacterial lipase. AB - In situ adsorption, known as an in situ-roduct removal (ISPR) technique for low molecular mass bioproducts, was in this study applied to a bacterial exoenzyme proving that this method is also suitable for the separation of macromolecules like proteins. For this, adsorbent particles were added to growing cultures of Staphylococcus carnosus rec., therefore both production and adsorption occurred simultaneously in shaking flasks, stirred tank, or airlift bioreactor as the chosen types of fermenters. The exoenzyme lipase adsorbed rapidly and, after separating cells and adsorbents, desorbed in a packed bed column. Up to 85% of the produced lipase were recovered, fractions of these had been concentrated up to the factor 20 and purified up to a factor of 40 by the procedure. By using the airlift bioreactor an enhancement of biomass production was observed, but the necessity of the addition of an anti-foam reagent resulted in higher product losses in adsorption as well as in desorption. Production and adsorption kinetics have been modeled and applied to in situ-adsorption. The model was used to perform a parameter study in which the influence of biological and physical parameters as well as process parameters on discontinuous and continuous in situ adsorption was investigated. PMID- 16267850 TI - Potential application of monolith packed columns as bioreactors, control of biofilm formation. AB - Monolith reactors combine good mass transfer characteristics with low-pressure drop, the principle factors affecting the cost effectiveness of industrial processes. Recently, these specific features of the monolith reactors have drawn the attention toward the application of the monolith reactor in multiphase reaction systems. In this study, we explore the potential application of monolith reactors as bioreactor requiring gas-liquid mass transfer for substrate supply. It is demonstrated on theoretical grounds that the monolith reactor is a competitive alternative to conventional gas-liquid bioreactors such as stirred tanks, packed beds, and airlift bioreactors because it allows for a significant reduction of the energy dissipation that is normally required for gas-liquid contacting. A potential problem of monolith reactors for biological processes is clogging due to biofilm formation. This paper presents experimental results of a study into the formation and possible removal of biofilms during operation of a monolith reactor as suspended cells bioreactor. The results indicate that biofilm formation may be minimized and postponed by a proper choice of operating conditions. Periodic biofilm removal could straightforwardly be achieved by rinsing with water at moderate pressures and allows for stable operation for prolonged periods of time. PMID- 16267852 TI - Technological advances in MRI measurement of brain perfusion. AB - Measurement of brain perfusion using arterial spin labeling (ASL) or dynamic susceptibility contrast (DSC) based MRI has many potential important clinical applications. However, the clinical application of perfusion MRI has been limited by a number of factors, including a relatively poor spatial resolution, limited volume coverage, and low signal-to-noise ratio (SNR). It is difficult to improve any of these aspects because both ASL and DSC methods require rapid image acquisition. In this report, recent methodological developments are discussed that alleviate some of these limitations and make perfusion MRI more suitable for clinical application. In particular, the availability of high magnetic field strength systems, increased gradient performance, the use of RF coil arrays and parallel imaging, and increasing pulse sequence efficiency allow for increased image acquisition speed and improved SNR. The use of parallel imaging facilitates the trade-off of SNR for increases in spatial resolution. As a demonstration, we obtained DSC and ASL perfusion images at 3.0 T and 7.0 T with multichannel RF coils and parallel imaging, which allowed us to obtain high-quality images with in-plane voxel sizes of 1.5 x 1.5 mm(2). PMID- 16267853 TI - Arterial spin labeling in small animals: methods and applications to experimental cerebral ischemia. AB - ASL enables noninvasive, quantitative monitoring of cerebral perfusion to be performed repeatedly over a period of hours. Thus, ASL is an attractive method for basic science studies of the time evolution and pathophysiology of diseases using animal models. In particular, ASL is valuable for basic science studies of evolving tissue status and viability in stroke using animal models of acute ischemia. This study describes both pulsed (PASL) and continuous ASL (CASL) studies of quantitative cerebral perfusion in rodent models of cerebral ischemia. Some technical factors pertinent to these studies are discussed, including a method for measuring arterial blood T(1) and double-echo PASL for measuring cerebral blood flow (CBF) and volume (CBV). Investigations of the CBF response to forebrain ischemia and reperfusion, and of regional variations in CBF and arterial transit time (ATT) are also discussed. PMID- 16267854 TI - Quantification of cerebral perfusion using arterial spin labeling: two compartment models. AB - One of the advantages of arterial spin labeling (ASL) techniques over other techniques for measuring cerebral perfusion is that with ASL it is possible to achieve accurate quantification. This is particularly useful in the field of functional imaging, where accurate measurements of perfusion change can help untangle the complex physiological changes that occur following neuronal activation. However, the linearity of the perfusion estimate over a wide range of perfusion values may be more important than absolute values. For several years, single-compartment models have dominated the literature, and it has been assumed that the labeled water diffuses freely throughout the tissue voxel. However, recent work, as summarized in this review, has shown that this assumption is inaccurate and leads to an overestimation of perfusion at low perfusion rates, and an underestimation at high rates. The inclusion of restricted permeability of the capillary wall to water in a two-compartment model offers improved quantification. PMID- 16267855 TI - Perfusion-based fMRI: insights from animal models. AB - Modern functional neuroimaging techniques, including functional magnetic resonance imaging (fMRI), positron emission tomography (PET), and optical imaging of intrinsic signals (OIS), rely on a tight coupling between neural activity and cerebral blood flow (CBF) to visualize brain activity using CBF as a surrogate marker. Because CBF is a uniquely defined physiological parameter, fMRI techniques based on CBF contrast have the advantage of being specific to tissue signal change, and the potential to provide more direct and quantitative measures of brain activation than blood oxygenation level-dependent (BOLD)- or cerebral blood volume (CBV)-based techniques. The changes in CBF elicited by increased neural activity are an excellent index of the magnitude of electrical activity. Increases in CBF are more closely localized to the foci of increased electrical activity, and occur more promptly to the stimulus than BOLD- or CBV-based contrast. In addition, CBF-based fMRI is less affected by confounds from venous drainage common to BOLD. Animal studies of brain activation have yielded considerable insights into the advantages of CBF-based fMRI. Based on results provided by animal studies, CBF fMRI may offer a means of better assessing the magnitude, spatial extent, and temporal response of neural activity, and may be more specific to tissue state. These properties are expected to be particularly useful for longitudinal and quantitative fMRI studies. PMID- 16267856 TI - The effect of chronic inflammatory condition of the bladder and estrogen replacement therapy on bladder functions and histology in surgically menopause and chronic cystitis induced rats. AB - AIMS: We investigated the effect of chronic inflammatory condition of the bladder and estrogen replacement therapy on bladder function and histological changes in surgically menopause and chronic cystitis induced rats. METHODS: The study included 36 female Sprague-Dawley rats, divided into five groups. After bilateral ovariectomy, chronic cystitis was induced by intravesical instillation of HCl, and then group 1 (n = 8) received nasal estrogen, and group 2 (n = 8) received oral estrogen replacement therapy. Group 3 (n = 7) underwent ovariectomy and chronic cystitis, but no replacement therapy. Group 4 (n = 7) had only ovariectomy. Group 5 (n = 6) was taken as sham group. The rats were sacrificed after 60 days, and cystometric study and histological findings were compared among the groups. RESULTS: The mean maximal bladder capacity and compliance revealed significant decreases in group 3 and 4 compared with group 5, and significant increases in group 1 and 2 compared with group 3. Histological findings showed significant increases in the mast cells and leukocyte infiltration of group 3 and 4 compared with group 5, and significant decreases in the mast cells and leukocyte infiltration of group 1 and 2 compared with group 3. CONCLUSIONS: This experimental menopause and chronic cystitis model showed that bladder function and histology might deteriorate much more with chronic cystitis in postmenopausal period. This is the first study in the literature to report that chronic inflammatory bladder occurring after menopause can be improved by estrogen replacement therapy. PMID- 16267857 TI - Altered expression of thin filament-associated proteins in hypertrophied urinary bladder smooth muscle. AB - AIMS: Obstruction of the urinary bladder outlet induces detrusor smooth muscle (DSM) hypertrophy. The goal of this study was to determine whether the composition of thin filament-associated proteins, known to play important roles in cytoskeletal structure and/or the regulation of contraction, is altered in DSM during hypertrophy. METHODS: DSM hypertrophy was induced in male rabbits by partial ligation of the urethra. Sham-operated rabbits served as a control. Reverse transcriptase-polymerase chain reaction (RT-PCR) and real-time PCR revealed a significant increase in the expression of mRNAs for basic (h1) calponin (CaP), and alpha-isoform of tropomyosin (Tm) in hypertrophied DSM compared to controls. Western blotting and two-dimensional (2-D) gel electrophoresis showed enhanced expression of these proteins and also a significant increase in the expression of beta-non muscle and gamma-smooth muscle actin in the DSM from obstructed bladders, while alpha-actin remained constant. RESULTS: Enhanced expression of these proteins in the DSM from obstructed bladders was confirmed by immunofluorescence microscopy. Double immunostaining with Cap/Tm and alpha/beta-actin-specific antibodies showed co-localization of these proteins in myocytes. Colocalization of smooth muscle specific myosin and CaP to cytoplasmic filaments in cells dissociated from the hypertrophied DSM indicated that these cells are differentiated smooth muscle cells. CONCLUSIONS: The change in the isoforms of actin, Cap, and Tm may be part of the molecular mechanism for bladder compensation in increased urethral resistance. PMID- 16267858 TI - A combined urodynamic and electrophysiological study of diabetic cystopathy. AB - AIMS: The aim of this study is to compare urodynamics and electrophysiological studies in the diagnosis of diabetic cystopathy. MATERIALS AND METHODS: In this prospective study are included twenty six patients with diabetes mellitus diagnosed at least since 5 years; 17 patients with diabetes type II and 9 with type I. They were divided in two groups in respect to lower urinary tract symptoms (LUTS) and signs suggestive of lower urinary tract dysfunction (LUTD) according to the ICS standardization. Patients with LUTS/LUTD were included in Group A and patients without LUTS/LUTD in Group B. Patients underwent different studies: urodynamic, somatosensory evoked potentials (SSEP) of tibial and pudendal nerves, bulbocavernosus reflex (BCR), bulbocavernosus muscles' electromyography and motor evoked potentials after transcranial magnetic stimulation to indirectly investigate bladder's innervation. RESULTS: Abnormal urodynamics were found in 13 patients of group A (92.9%) and in 5 of group B (47.1%). This difference was statistically significant (P=0.009). Abnormally prolonged latency of P40 of tibial SSEP was found in 11 patients of group A (78.6%) and in 4 of group B (33.3%) and this difference was also statistically significant (P=0.04). Differences between the two groups concerning: i) peripheral polyneuropathy; ii) pudendal SSEP, iii) dysfunction of central nervous system and iv) abnormal BCR were not statistically significant. CONCLUSION: The study of tibial SSEP is an easily performed test and it is well correlated to abnormal urodynamics in diabetic patients with and without LUTD/LUTS. PMID- 16267859 TI - Aurora B expression directly correlates with prostate cancer malignancy and influence prostate cell proliferation. AB - BACKGROUND: Chromosomal instability is one of the most common features of prostate cancer (PC), especially in advanced stages. Recent studies suggest that defects in mitotic checkpoints play a role in carcinogenesis. Lack of mitotic regulation induces aneuploidy in cancer cells acting thereafter as a driving force for malignant progression. Serine/threonine protein kinases of the Aurora genes family play an important throughout the entire cell cycle. In that Aurora B regulates chromosome segregation by ensuring the orientation of sister chromatids. As a consequence, the overexpression of Aurora B in diploid human cells NHDF induces the appearance of multinucleate cells. METHODS: Archive samples of normal and neoplastic prostate tissue, and prostate derived cell lines were screened for the expression of Aurora B. RESULTS: Immunohistochemical analysis showed increased nuclear expression of Aurora-B in high Gleason grade PCs respect to low and intermediate grade cases and in all cancers in respect to hyperplastic and normal glands. Furthermore, in the high Gleason grade anaplastic cancer tissues Aurora B expression was accompanied by the phosphorylation of the histone H3. In analogy to the in vivo situation, Aurora B was vigorously expressed in the androgen independent PC cell lines PC3 and DU145, while a very modest expression of the kinase was observed in the androgen sensitive LnCap cells and in the EPN cells, a line of epithelial cells derived from normal prostate tissue. In addition, in PC3 cells Aurora B expression is accompanied the by the phosphorylation of the histone H3. The block of Aurora B expression induced by an inhibitor of Aurora kinase activity significantly reduced the growth of prostate carcinoma cells, but not that of non-transformed EPN cells. CONCLUSIONS: Our data are the first demonstration of a role of Aurora B in PC progression. In addition, the observation that Aurora B specific inhibitors interfere with PC cell proliferation but not with that of non-transformed prostate epithelial cells suggest that Aurora B is a potential therapeutic target for PC. PMID- 16267860 TI - (S)-selective kinetic resolution and chemoenzymatic dynamic kinetic resolution of secondary alcohols. AB - (S)-Selective kinetic resolution was achieved through the use of a commercially available protease, which was activated with a combination of two different surfactants. The kinetic resolution (KR) process was optimized with respect to activation of the protease and to the acyl donor. The KR proved to be compatible with a range of functionalized sec-alcohols, giving good to high enantiomeric ratio values (up to >200). The enzymatic resolution was combined with a ruthenium catalyzed racemization to give an (S)-selective dynamic kinetic resolution (DKR) of sec-alcohols. The DKR process works under very mild reaction conditions to give the corresponding esters in high yields and with excellent enantioselectivities. PMID- 16267861 TI - Total synthesis of phorboxazole B. AB - An efficient and highly convergent total synthesis of the potent antitumor agent phorboxazole B has been achieved. The synthetic strategy of this synthesis features: 1) a highly efficient substrate-controlled hydrogenation to construct the functionalized cis-tetrahydropyrane unit; 2) iterative crotyl addition to synthesize the segment that contains alternating hydroxyl and methyl substituents; 3) Hg(OAc)2/I2-induced cyclization to establish the cis tetrahydropyrane moiety; 4) 1,3-asymmetric induction in the Mukaiyama aldol reaction to afford the stereogenic centers at C9 and C3; and 5) the exploration of the Still-Gennari olefination reaction to complete the macrolide ring of phorboxazoloe B. PMID- 16267862 TI - NMR investigation of the dihydrogen-bonding and proton-transfer equilibria between the hydrido carbonyl anion [HRe2(CO)9]- and fluorinated alcohols. AB - The interaction of fluorinated alcohols with the anionic hydrido complex [HRe2(CO)9]- (1) has been investigated by NMR spectroscopy. According to the acidic strength of the alcohols, the interaction may result not only in the formation of dihydrogen-bonded ROH...[HRe2(CO)9]- adducts 2, but also in proton transfer to give the neutral species [H2Re2(CO)9] (3). With the weaker acid trifluoroethanol (TFE) evidence for the occurrence of the dihydrogen-bonding equilibrium was obtained by 2D 1H NOESY. The dependence of the hydride chemical shift on TFE concentration at different temperatures provided values for the constants of this equilibrium, from which the thermodynamic parameters were evaluated as deltaH(degrees) = -2.6(2) kcal mol(-1), deltaS(degrees) = -9.3(2) cal mol(-1) K(-1). This corresponds to a rather low basicity factor (E(j) = 0.64). Variable-temperature T1 measurements allowed the proton-hydride distance in adduct 2 a to be estimated (1.80 angstroms). In the presence of hexafluoroisopropyl alcohol (HFIP) simultaneous occurrence of both dihydrogen bonding and proton-transfer equilibria was observed, and the equilibria shifted versus the protonated product 3 with increasing HFIP concentration and decreasing temperature. Reversible proton transfer between the alcohol and the hydrido complex occurs on the NMR timescale, as revealed by a 2D 1H EXSY experiment at 240 K. For the more acidic perfluoro-tert-butyl alcohol (PFTB) the protonation equilibrium was further shifted to the right. Thermal instability of 3 prevented the acquisition of accurate thermodynamic data for these equilibria. The occurrence of the proton-transfer processes (in spite of the unfavorable pK(a) values) can be explained by the formation of homoconjugated RO...HOR- pairs which stabilize the alcoholate anions. PMID- 16267863 TI - Nitrogen fixation under mild ambient conditions: part I--the initial dissociation/association step at molybdenum triamidoamine complexes. AB - In several recent studies Schrock and collaborators demonstrated for the first time how molecular dinitrogen can be catalytically transformed under mild and ambient conditions to ammonia by a molybdenum triamidoamine complex. In this work, we investigate the geometrical and electronic structures involved in this process of dinitrogen activation with quantum chemical methods. Density functional theory (DFT) has been employed to calculate the coordination energies of ammonia and dinitrogen relevant for the dissociation/association step in which ammonia is substituted by dinitrogen. In the DFT calculations the triamidoamine chelate ligand has been modeled by a systematic hierarchy of increasingly complex substituents at the amide nitrogen atoms. The most complex ligand considered is an experimentally known ligand with an HMT = 3,5-(2,4,6-Me3C6H2)2C6H3 substituent. Several assumptions by Schrock and collaborators on key reaction steps are confirmed by our calculations. Additional information is provided on many species not yet observed experimentally. Particular attention is paid to the role of the charge of the complexes. The investigation demonstrates that dinitrogen coordination is enhanced for the negatively charged metal fragment, that is, coordination is more favorable for the anionic metal fragment than for the neutral species. Coordination of N2 is least favorable for the cationic metal fragment. Furthermore, ammonia abstraction from the cationic complex is energetically unfavorable, while NH3 abstraction is less difficult from the neutral and easily feasible from the anionic low-spin complex. PMID- 16267865 TI - A theoretical investigation into the photophysical properties of ruthenium polypyridine-type complexes. AB - Excited states of ruthenium polypyridine-type complexes have always attracted the interest of chemists. We have recently found evidence of a remarkable long-lived excited state (30 micros) for a Ru(II) complex containing a heteroditopic ligand that can be viewed as a fused phenanthroline and salophen ligand.1 To unravel this intriguing electronic property, we have used density functional theory (DFT) calculations to understand the ground-state properties of [(bpy)(2)Ru(LH(2))](2+), where LH(2) represents N,N'-bis(salicylidene)-(1,10 phenanthroline)diamine. Excited singlet and triplet states have been examined by the time-dependent DFT (TDDFT) formalism and the theoretical findings have been compared with those for the parent complex [Ru(bpy)(3)](2+). The outstanding result is the presence of excited states lower in energy than the metal-to-ligand charge-transfer states, originating from intraligand charge transfer (ILCT) from the phenolic rings to the phenanthroline part of the coordinated LH(2). The spin density distribution for the lowest triplet state provides evidence that it is in fact the lowest triplet state of the free ligand. Correlation between the energy level diagram of orbitals for the ground state and that for the (3)ILCT state clearly establishes that the ruthenium retains its formal Ru(II) oxidation state. The quenching of the luminescence and the evidence of the long-lived excited state observed for [(bpy)(2)Ru(LH(2))](2+) are discussed in the light of the computational results. PMID- 16267864 TI - Relaxometric study of copper [15]metallacrown-5 gadolinium complexes derived from alpha-aminohydroxamic acids. AB - Proton nuclear magnetic relaxation dispersion (NMRD) profiles were recorded between 0.24 mT and 1.4 T for lanthanum(III)- and gadolinium(III)-containing [15]metallacrown-5 complexes derived from alpha-aminohydroxamic acids and with copper(II) as the ring metal. The influence of the different R-groups on the proton relaxivity was investigated, and a linear relationship between the relaxivity and the molecular mass of the metallacrown complex was found. The selectivity of the metallacrown complexes was tested by transmetalation experiments with zinc(II) ions. The crystal structure of the copper [15]metallacrown-5 gadolinium complex with glycine hydroximate ligands is reported. PMID- 16267866 TI - Carbohydrate chain of ganglioside GM1 as a ligand: identification of the binding strategies of three 15 mer peptides and their divergence from the binding modes of growth-regulatory galectin-1 and cholera toxin. AB - The branched pentasaccharide chain of ganglioside GM1 is a prominent cell surface ligand, for example, for cholera toxin or tumor growth-regulatory homodimeric galectins. This activity profile via protein recognition prompted us to examine the binding properties of peptides with this specificity. Our study provides insights into the mechanism of molecular interaction of this thus far unexplored size limit of the protein part. We used three pentadecapeptides in a combined approach of mass spectrometry, NMR spectroscopy and molecular modelling to analyze the ligand binding in solution. Availability of charged and hydrophobic functionalities affected the intramolecular flexibility of the peptides differently. Backfolding led to restrictions in two cases; the flexibility was not reduced significantly by association of the ligand in its energetically privileged conformations. Major contributions to the interaction energy arise from the sialic acid moiety contacting Arg/Lys residues and the N-terminal charge. Considerable involvement of stacking between the monovalent ligand and aromatic rings could not be detected. This carbohydrate binding strategy is similar to how an adenoviral fiber knob targets sialylated glycans. Rational manipulation for an affinity enhancement can now be directed to reduce the flexibility, exploit the potential for stacking and acquire the cross-linking capacity of the natural lectins by peptide attachment to a suitable scaffold. PMID- 16267867 TI - Hydrogen bonding and dynamic behaviour in crystals and polymorphs of dicarboxylic diamine adducts: a comparison between NMR parameters and X-ray diffraction studies. AB - Fumaric, malonic, maleic, and hydromuconic (HOOCCH2(CH)2CH2COOH) acids were used to prepare a series of hydrogen-bonded adducts or salts, depending on whether acid-base proton transfer takes place, with the dibase [N(muCH2CH2)3N] in various stoichiometric ratios. The resulting compounds have been investigated by using the 1H MAS, 15N, and 13C cross polarisation magic-angle spinning (CPMAS) methods and discussed in relation to X-ray diffraction studies to ascertain the nature of the O-HO, NH-O, and N+-HO- hydrogen bonds between the various species. In addition, two polymorphic forms of the malonic compound and a hydrate in the maleic case were examined. We also present the correlations between the chemical shifts of the hydrogen-bonded protons and those from the proton transfer reaction (acid-to-base) with the heavy atom distances. The dynamic behaviour in the solid state of the [N(muCH2CH2)3N] adducts with fumaric 2:1, maleic 1:1 hydrate, and hydromuconic acids, and a malonate 2:1 polymorph adduct have been investigated by using variable-temperature 1H spin-lattice relaxation times. A substantial agreement between the activation energies obtained from fitting the T1 data and the results of potential energy barrier calculations demonstrates that the facile reorientation of the [N(muCH2CH2)3N] molecule occurs in several of the adducts. PMID- 16267868 TI - Zinc hydrazides and alkoxyhydrazides: organometallic compounds with novel Zn4N8, Zn4N6O and Zn4N4O2 cage structures. AB - Tetrameric [{RZn(NHNMe2)}4] (R = Me, Et), the first organometallic zinc hydrazides to be described, have been prepared by alkane elimination from dialkylzinc solutions and N,N-dimethylhydrazine. They were characterised by 1H and 13C NMR and IR spectroscopy, mass spectrometry, elemental analysis and X-ray crystallography. The compounds form asymmetric aggregates containing the novel Zn4N8 core; tetrahedra of Zn atoms bear the alkyl groups at Zn, with the triangular faces bridged by NHNMe2 substituents. The NH groups are connected to two Zn atoms, and the NMe2 groups to one. Hydrolysis of the compounds with water gives [(RZn)4(OH)(NHNMe2)3] as products, which also were characterised as described above. Higher yields of these hydroxo clusters were achieved in one-pot syntheses by reaction of dialkylzinc solutions with mixtures of N,N dimethylhydrazine and water. They contain Zn4N6O cages, in which one hydroxide in the tetrameric hydrazides described above replaces one NHNMe2 group. Similar products can be prepared with alkoxy instead of hydroxy groups, in analogous one pot syntheses with alcohols. Alcoholysis of [EtZn(NHNMe2)]4 with methanol or ethanol gave zinc trishydrazide monoalkoxides, [(EtZn)4(OR)(NHNMe2)3] (R = Me, Et), which have constitutions analogous to the monohydroxides. The organozinc bishydrazide bisalkoxides [(MeZn)4(NHNMe2)2(OEt)2] and [(EtZn)4(NHNMe2)2(OEt)2] were obtained in one-pot reactions from dialkylzinc solutions with mixtures of the hydrazine and alcohol, and their crystal structures, confirmed by spectroscopic methods in solution, show an unsymmetrical aggregation with the novel Zn4N4O2 cage structure. PMID- 16267869 TI - Alkynylpyrenes as improved pyrene-based biomolecular probes with the advantages of high fluorescence quantum yields and long absorption/emission wavelengths. AB - The photochemical properties of various alkynylpyrene derivatives have been investigated in detail with a view to developing a new class of pyrene-based biomolecular probes. The absorption maxima of the alkynylpyrenes were seen to be shifted to longer wavelengths compared with those of the unsubstituted parent pyrene. Fluorescence quantum yields of the alkynylpyrenes dramatically increased up to 0.99 in ethanol, and only slight quenching of the fluorescence occurred even under aerated conditions. The alkynylpyrenes have been successfully introduced into representative biomolecules such as peptides, proteins, and DNAs. The detectabilities of the labeled biomolecules were significantly improved, with the unique photochemical characteristics of the pyrene nucleus being maintained. PMID- 16267870 TI - Aminolysis of 2,4-dinitrophenyl X-substituted benzoates and Y-substituted phenyl benzoates in MeCN: effect of the reaction medium on rate and mechanism. AB - Second-order rate constants (k(N)) have been determined spectrophotometrically for the reactions of 2,4-dinitrophenyl X-substituted benzoates (1 a-f) and Y substituted phenyl benzoates (2 a-h) with a series of alicyclic secondary amines in MeCN at 25.0 +/- 0.1 degrees C. The k(N) values are only slightly larger in MeCN than in H2O, although the amines studied are approximately 8 pK(a) units more basic in the aprotic solvent than in H2O. The Yukawa-Tsuno plot for the aminolysis of 1 a-f is linear, indicating that the electronic nature of the substituent X in the nonleaving group does not affect the rate-determining step (RDS) or reaction mechanism. The Hammett correlation with sigma- constants also exhibits good linearity with a large slope (rho(Y) = 3.54) for the reactions of 2 a-h with piperidine, implying that the leaving-group departure occurs at the rate determining step. Aminolysis of 2,4-dinitrophenyl benzoate (1 c) results in a linear Bronsted-type plot with a beta(nuc) value of 0.40, suggesting that bond formation between the attacking amine and the carbonyl carbon atom of 1 c is little advanced in the transition state (TS). A concerted mechanism is proposed for the aminolysis of 1 a-f in MeCN. The medium change from H2O to MeCN appears to force the reaction to proceed concertedly by decreasing the stability of the zwitterionic tetrahedral intermediate (T+/-) in aprotic solvent. PMID- 16267871 TI - Catalytic asymmetric mannich-type reactions activated by ZnF2 chiral diamine in aqueous media. AB - Catalytic asymmetric Mannich-type reactions of an alpha-hydrazono ester with silicon enolates in aqueous media have been developed by using ZnF2 and chiral diamines as catalysts. In these reactions, both Zn2+ and a fluoride anion were necessary to achieve high yields and enantioselectivities, suggesting a double activation mechanism, in which Zn2+ activates the alpha-hydrazono ester and the fluoride anion simultaneously activates the silicon enolate. When chiral diamine ligands bearing methoxy-substituted aromatic rings were employed, the reactions in aqueous THF were markedly accelerated. Furthermore, the use of these diamines facilitated the asymmetric Mannich-type reactions in water without any organic cosolvents. It is noteworthy that either syn or anti adducts were stereospecifically obtained from (E)- or (Z)-silicon enolates, respectively. Interestingly, these reactions proceeded smoothly only in the presence of water. On the basis of several experimental results, it can be concluded that the reaction mechanism is likely to be a fluoride-catalyzed one, in which the ZnF2 chiral diamine complex is regenerated from the Me3SiF formed during the reaction. PMID- 16267872 TI - Protein posttranslational modifications: the chemistry of proteome diversifications. AB - The diversity of distinct covalent forms of proteins (the proteome) greatly exceeds the number of proteins predicted by DNA coding capacities owing to directed posttranslational modifications. Enzymes dedicated to such protein modifications include 500 human protein kinases, 150 protein phosphatases, and 500 proteases. The major types of protein covalent modifications, such as phosphorylation, acetylation, glycosylation, methylation, and ubiquitylation, can be classified according to the type of amino acid side chain modified, the category of the modifying enzyme, and the extent of reversibility. Chemical events such as protein splicing, green fluorescent protein maturation, and proteasome autoactivations also represent posttranslational modifications. An understanding of the scope and pattern of the many posttranslational modifications in eukaryotic cells provides insight into the function and dynamics of proteome compositions. PMID- 16267873 TI - Short de novo synthesis of fully functionalized uronic acid monosaccharides. PMID- 16267874 TI - [MnIII(salen)]6[FeIII(bpmb)(CN)2]67 H2O: a cyanide-bridged nanosized molecular wheel. PMID- 16267875 TI - The effects of triplet sensitizers' energies on the photoreactivity of beta,gamma unsaturated methyl ketones. PMID- 16267876 TI - Finely tuning metallic nanogap size with electrodeposition by utilizing high frequency impedance in feedback. PMID- 16267877 TI - Characterization of a complex comprising a [Cu2(S2)2]2+ core: bis(mu-S2 2 )dicopperIII or bis(mu-S2).-)dicopperII? PMID- 16267878 TI - Scope for improvement in the quality of reporting of systematic reviews. From the Cochrane Musculoskeletal Group. AB - OBJECTIVE: To assess the quality of reporting in Cochrane musculoskeletal systematic reviews (excluding back and injury reviews). METHODS: This study assessed all the Cochrane Musculoskeletal Group's systematic reviews from Issue 4, 2002, of the Cochrane Library Database of Systematic Reviews. Two reviewers independently extracted data and assessed quality. Two assessment tools were used, including an 18 item checklist and flow chart developed by the Quality of Reporting of Meta-analysis (QUOROM) consensus group, and a 10 item scale, the Oxman-Guyatt Overview Quality Assessment Questionnaire (OQAQ). One question on the latter scale (item 10) scores overall quality on a 7 point scale, with high scores indicating superior quality. Data were analyzed using univariate approaches. RESULTS: The 57 systematic reviews assessed were found to have good overall quality, with scores on individual items revealing only minor flaws. Documenting the flow of included and excluded studies and summarizing the results are 2 areas needing improvement in reporting. According to the Oxman-Guyatt scale the overall scientific quality of the Cochrane musculoskeletal reviews was good [mean 5.02 (95% CI 3.71-6.32)]. CONCLUSION: Our study found that the reporting quality of Cochrane musculoskeletal systematic reviews was generally good, although there was room for improvement. For example, it might be feasible to develop specific guidelines for reporting protocols. Certainly more work is needed in reporting search results, documentation of the flow of studies, identification of the type of studies, and summarization of the key findings. PMID- 16267879 TI - Frequency and predictors of inappropriate management of recurrent gout attacks in a longitudinal study. AB - OBJECTIVE: To evaluate the patterns and determinants of medication use during recurrent gout attacks. METHODS: We followed participants with documented gout in an online prospective case-crossover study. During an attack, subjects were asked if they had consulted a physician for the attack and what medications they were using. Definitely inappropriate therapy was defined as use of allopurinol or a uricosuric agent acutely without having used it as a prophylactic. Potentially inappropriate therapy was defined as use of analgesics alone, alternative remedies, or no medications. We estimated the risk of having >or= 1 attack in 1 year using life table methods. We examined the relation of various risk factors to the risk of inappropriate therapy using Poisson regression. RESULTS: Among 232 participants (mean age 52 yrs, 81% male) with documented gout, the risk of having >or= 1 attack in a year was 69%. One hundred ten participants consulted a physician for each attack, 49 did so for only some attacks, while 43 never consulted a physician for any attack. Fifty-three participants had definitely (n = 10) or potentially (n = 43) inappropriate therapy for their recurrent attacks. Physician consultation for an attack was associated with increased risk of inappropriate therapy (risk ratio, RR, 2.5, p = 0.006), whereas an increasing number of gout attacks was associated with lower risk of inappropriate therapy (RR 0.8, p = 0.01). CONCLUSION: Given the high risk of recurrent attacks and the substantial number of persons whose attacks are not appropriately managed, further education about management of gout attacks for both patients and physicians may be warranted. PMID- 16267880 TI - Couple-focused support improves medication adherence. PMID- 16267881 TI - Commentary: nicotine replacement therapy for smoking cessation. PMID- 16267882 TI - Commentary; nicotine replacement therapy for smoking cessation. PMID- 16267883 TI - Stereochemistry of allene biosynthesis and the formation of the acetylenic carotenoid diadinoxanthin and peridinin (C37) from neoxanthin. PMID- 16267884 TI - 2005 Guidelines for the diagnosis, prevention and management of heartworm (Dirofilaria immitis) infection in dogs. PMID- 16267885 TI - 2005 Guidelines for the diagnosis, prevention and management of heartworm (Dirofilaria immitis) infection in cats. PMID- 16267886 TI - Approval of drug marketed to African Americans raises political, economic issues. Study criticizes manufacturer for re-branding generic drug with race-specific indication. PMID- 16267887 TI - Few middle-aged women with severe mental symptoms use psychotropic drugs: the Women's Health in Lund Area (WHILA) study. AB - AIMS: In a population of middle-aged women a survey was carried out to ascertain the prevalence of mental symptoms and psychotropic drug use, and further to investigate whether severe mental symptoms are associated with social situation, alcohol consumption, smoking, and physical health. METHODS: All women (n = 10,766) aged 50-59 years and living in the Lund area were invited to the WHILA study, a health survey including laboratory examinations and a self-administered questionnaire; 6,917 (64.2%) participated. This study is based on the questionnaire only. RESULTS: During the past three months 25.4% (n = 1,709) had been troubled by none or 1 mental symptom (labelled "absent/slight"), 52.8% (n = 3,555) by 2-6 mental symptoms ("moderate") and 21.8% (n = 1,471) by 7-10 mental symptoms ("severe"). Among women with severe mental symptoms 15.4% regularly used psychotropic drugs, mainly antidepressants. A multiple logistic regression analysis showed that women with severe mental symptoms had higher odds for living alone (OR 1.7; CI 1.3-2.2) or as single parents (OR 2.1; CI 1.2-3.6), being university-educated (OR 1.5; CI 1.1-2.0), being on long-term sick-leave (OR 8.8; CI 3.0-25.5), using hormone replacement therapy (OR 1.3; CI 1.1-1.6), and having severe physical symptoms (136.8; CI 89.2-209.7) compared with women with absent/slight mental symptoms. CONCLUSION: Mental symptoms were common among the participating women. The presence of severe mental symptoms was strongly associated with severe physical symptoms. Few women with severe mental symptoms used psychotropic drugs. Middle-aged women with severe mental symptoms need to be identified and provided with appropriate psychopharmacological, hormonal, and/or psychosocial treatment. PMID- 16267888 TI - Preventive home visits to older people in Southern Sweden. AB - BACKGROUND: It seems urgent to further develop and modify models of preventive measures in order to prolong a healthy functional and social situation for older people, as this population is increasing globally. The aim of the study was to describe a Swedish model of preventive home visits for persons aged 78 years with the focus on the effect on physical and social well-being, and the participants' and the visitors' experience of the home visits. METHOD: Three annual visits were performed and included 150 persons, 78 years old at the first visit. Data were collected in the form of a structured interview with the persons who received the visits and as a questionnaire for their visitors. RESULTS: Differences over time could be seen for the participants' physical and social activity, indicating that there was a deterioration between year one and two that was not seen in year three. The results further showed that the participants felt more secure and well informed as a result of the visits. The visitors said that they had gained a new, more positive view of older people and increased job satisfaction as a result of the visits and emphasized the impact on their own professional development. CONCLUSION: As no control group was used, the interpretation of effects concerning health factors has to be interpreted with caution. The visits did, however, have a positive impact on the participants and the visitors. PMID- 16267889 TI - Myths and perceptions of back pain in the Norwegian population, before and after the introduction of guidelines for acute back pain. AB - AIM: Deyo's seven "myths" about back pain are based on common misconceptions of causality and therapy of back pain. These myths were alive in the Norwegian population in 2001; this report investigates whether this is true two years later. METHODS: A representative sample of the Norwegian population in 2003 (n = 1,014) were asked to rate their agreement with the seven myths. RESULTS: There is significantly less acceptance of all myths except "Most back pain is caused by injuries or heavy lifting" in 2003. Myths concerning the use and importance of X ray were still common: 43% agreed that "X-ray and newer imaging tests can always identify the cause of pain" and 50% that "Everyone with back pain should have a spine X-ray". Low level of education is still associated with high acceptance of the myths, but the changes from 2001 are most pronounced for these groups. CONCLUSION: Perceptions in the general population in Norway are slowly changing to be more in accordance with existing knowledge on accurate behaviour and treatment of back pain. This may be related to introduction of new evidence-based guidelines. The difference between educational groups is reduced, but is still a challenge to health professionals and health authorities. PMID- 16267890 TI - [Image of the month: ulcerated mammary cancer]. PMID- 16267891 TI - [Anemia by self-injury: the Lasthenie de Ferjol syndrome]. AB - Patients with factitious disorders need to be ill and to defy physicians. These syndromes are difficult to diagnose because of the permanent disorder appearing inside the therapeutic relation. The case of a young woman who presents with a factitious anemia, also called Lasthenie de Ferjol syndrome, shows the complexity of such psychosomatic symptoms. PMID- 16267892 TI - [The hemolytic uremic syndrome: prime reason for acute renal failure in children]. AB - The Hemolytic Uremic Syndrome (HUS) is the prime cause for acute renal failure in children. The HUS is a combination of hemolytic anemia, thombopenia and acute nephropathy: all signs of a thrombotic microangiopathy. Onset occurs generally in infancy and is often associated with severe bloody diarrhea. Most of those cases are caused by Escherichia coli O157:H7 witch produces an exotoxin responsible for the microangiopathy. We discuss the treatment of HUS based on the experience acquired since 1994 in our Paediatric Intensive Care Unit (PICU), University of Liege. The frequent association of dehydration, multi-systemic impairment and reno-vascular hypertension justifies the early admission for PICU-surveillance. This allows the difficult fluid balance management in a setting of renal and pre renal failure. PMID- 16267893 TI - On interferon research. PMID- 16267894 TI - Euthanasia versus letting die: Christian decision-making in terminal patients. AB - Utilitarianism and quality-of-life considerations have increased the pressure to devalue life in terminal situations, leading to ethical confusion among caregivers. Where is the balance between a commitment to life and a commonsense willingness to "let go" when the time comes? This paper explores this balance, using a case history of a man with respiratory failure. This provides an opportunity to define and discuss some commonly misunderstood concepts related to end-of-life care. The ethical principles of terminal care are presented from the viewpoint of both secular and Christian ethics. PMID- 16267895 TI - Whistleblowing in the health-related professions. PMID- 16267897 TI - Doctors and the plague. PMID- 16267896 TI - Medical ethics in India: ancient and modern (I). PMID- 16267898 TI - Ethical problems in medical education. PMID- 16267899 TI - Urinary tract infections. PMID- 16267900 TI - Commercial support for continuing medical education. PMID- 16267901 TI - Human values in genetics and embryo experiments. PMID- 16267902 TI - Patients testing positive for HIV--ethical dilemmas in India. PMID- 16267904 TI - Biotechnology and medicine: ethical concerns. PMID- 16267903 TI - Medical ethics: relationships between doctors. PMID- 16267905 TI - Fraud in medical research. PMID- 16267906 TI - Cover illustration: a choroidal sleight of hand. PMID- 16267907 TI - Clinicians prescribing exercise: is air pollution a hazard? PMID- 16267908 TI - Body mass index predicts the incidence of radiation pneumonitis in breast cancer patients. AB - In patients receiving breast radiotherapy, the risk of radiation pneumonitis has been associated with the volume of irradiated lung, and concomitant methotrexate, paclitaxel, and tamoxifen therapy. Many of the studies of radiation pneumonitis are based on estimates of pulmonary risk using central lung distance that is calculated using two-dimensional techniques. With the treatment of internal mammary nodes and three-dimensional treatment planning for breast cancer becoming increasingly more common, there is a need to further consider the impact of dose volume metrics in assessing radiation pneumonitis risk. We herein present a case control study assessing the impact of clinical and dose-volume metrics on the development of radiation pneumonitis in patients receiving sequential chemotherapy and local-regional radiotherapy. PMID- 16267909 TI - Parental drift. PMID- 16267911 TI - [Risk management--adverse effects of pharmaceutical products. Interpretation of the drug package inserts]. PMID- 16267910 TI - Terror firmer. PMID- 16267912 TI - Farming for new opportunities. PMID- 16267913 TI - Skirting around thin ice. PMID- 16267914 TI - Susan Gasser. Interview. PMID- 16267915 TI - Surrogacy from a feminist perspective. PMID- 16267916 TI - Surrogacy and human reproductive biology. PMID- 16267917 TI - Sponsorships for medical specialists. PMID- 16267918 TI - Doctor-doctor ethics. PMID- 16267919 TI - Sponsored medical education? PMID- 16267920 TI - Taking a second look: some opinions on the second opinion. PMID- 16267921 TI - The cloning bandwagon: current discussion on the ethical consequences of genetic research is misplaced. PMID- 16267922 TI - Whose body is it anyway? An updated model of healthcare decision-making rights for adolescents. PMID- 16267923 TI - Physician, do no harm. PMID- 16267924 TI - Antiretrovirals in India. PMID- 16267926 TI - The 17th Annual Meeting of the Winter Conference on Neural Plasticity, Guadeloupe, French West Indies, February 19-26, 2005. PMID- 16267925 TI - Ethics, rights and strike of health workers. PMID- 16267928 TI - Potential CNS Drugs Experimental Biology 2005 Meeting, San Diego, California, USA, April 2-6, 2005. PMID- 16267927 TI - Medical ethics and medical education: some thoughts. PMID- 16267929 TI - Questions and miniscrews. PMID- 16267930 TI - Teaching medical ethics: a model. PMID- 16267931 TI - Privatisation of health care: new ethical dilemmas. PMID- 16267932 TI - HIV and confidentiality in India. PMID- 16267934 TI - The cloning bandwagon: a hysterical outburst. PMID- 16267933 TI - The ethics of cloning. PMID- 16267935 TI - The medical profession and human rights. PMID- 16267936 TI - Sexual assault: the role of the examining doctor. PMID- 16267937 TI - The medical profession and human rights. PMID- 16267939 TI - Mechanisms in heart, lung, and vessel diseases. Selected articles from the 1st International Conference on Recent Advances in Biomedical and Therapeutic Sciences, January 13-15, 2004, Jhansi, India. PMID- 16267938 TI - The ethics of public health: how can equity, social justice and human rights be incorporated into public health programmes? PMID- 16267940 TI - Practising ethically in a high-tech specialty. PMID- 16267941 TI - Iatrogenic error and truth telling: a comparison of the United States and India. PMID- 16267943 TI - The patient with AIDS. PMID- 16267942 TI - The pharmaceutical industry and the medical profession. PMID- 16267944 TI - Microvolt T-wave alternans testing to risk stratify patients being considered for ICD therapy for primary prevention of sudden death. PMID- 16267945 TI - Removing the uterus from mentally handicapped women: some ethical considerations. PMID- 16267946 TI - Suggested guidelines for hysterectomy in mentally handicapped women. PMID- 16267947 TI - The patient with AIDS--a response. PMID- 16267948 TI - Ethics in human medical research: views of a non-medical-person. PMID- 16267949 TI - Magnetic resonance-guided focused ultrasound therapy for symptomatic uterine fibroids. PMID- 16267950 TI - Hysterectomy in the mentally handicapped: an abridged version of the statement issued by PARYAY. PMID- 16267951 TI - Medical ethics--as prescribed by Caraka, Susruta and other ancient Indian physicians. PMID- 16267952 TI - Ethics of organ transplantation. PMID- 16267953 TI - Doctor-patient relationship. PMID- 16267954 TI - AIDS: Indian Council of Medical Research (ICMR) guidelines. PMID- 16267955 TI - Brainism: 'we are not our brains; we are embodied persons'. PMID- 16267956 TI - [Development of an antigen detection dot blot assay for the diagnosis of human onchocerciasis based on the biotin-avidin binding system]. AB - The development of a highly sensitive and specific diagnostic test is of urgent need for the field assessment of human onchocerciasis and for monitoring the success of control programs. We report here the development and evaluation of a Dot blot Immunobinding Assay (DIA-BA) based on the biotin-avidin binding system, for the detection of O. volvulus specific antigens in body fluids. Specific antibodies were produced by immunizing rabbits with the O. volvulus recombinant antigen Oncho-C71 and labelled with biotin. The biotinylated probes were then used to detect O. volvulus specific antigens initially blotted onto a nitrocellulose membrane. The smallest amount of blotted antigens detectable by the new test is 0.5ng, 1ng, 1ng and 2ng respectively in urine, dermal fluid, tears and serum samples. Out of 456 onchocerciasis endemic subjects examined, 98.4%, 96.5%, 90.8% and 75.0% were positive by the DIA-BA test on urine, dermal fluid, tears and serum respectively The test was most sensitive (100%) when used on urine and least (54.76%) when used on serum from skin snip positive subjects. The specificity of the test, determined amongst non-exposed individuals, was 100% on all but for dermal fluid samples (97.5%). Also, the color intensities on the blot were observed to positively correlate (r = 0.8 on urine) with the skin microfilaria loads on the individuals. We conclude that DIA-BA test could be very useful for mass diagnosis of prepatent, of low and high level infections due to O. volvulus. PMID- 16267957 TI - [Diagnosis and treatment of invasive cervical cancer in Cambodia (apropos of 35 cases)]. AB - Uterine cervix cancer is an important public health problem in developing countries. In Cambodia, incidence (24. 10-5) and mortality (12,77. 10-5) rates due to this cancer are higher than in other southeast Asian countries. 35 consecutive women with a diagnosis of invasive cervical cancer were identified from the histology laboratory of the Institut Pasteur in Phnom Penh. Information on history, clinical findings, ancillary investigations and treatment were obtained by clinical files review. Patients were from Phnom Penh, the main city and from outlying provinces. The mean age was 50 years, 2 months. They all presented with clinical symptoms at the time of diagnosis, and lesions had spread beyond the uterus in nearly half of the cases. Squamous cell carcinoma (80%) was more frequent than adenocarcinoma (14%). Microinvasive squamous cell carcinomas were rare (2.8%). Treatment was mainly surgical, but lacked standardization, as radical hysterectomy including lymphadenectomy was not systematic, even when the tumor spread beyond the uterine cervix. Few women that required adjuvant radiotherapy received it as this technique has only recently been introduced in Cambodia. Many patients were lost to follow-up. None of the cases was detected by pap smear. This technique is not in general use in the country. Cervical cancer screening could be carried out by visual inspection of the cervix with acetic acid wash that is less expensive and nearly as sensitive as pap smear. Treatments are still insufficiently standardized, due to the lack of technical means, and lack of patient compliance. PMID- 16267958 TI - [Dermatologic manifestations associated with immune reconstitution syndrome in HIV+ patients starting HAART: a retrospective study in French Guiana]. AB - Immune reconstitution syndrome (IRIS) is an unusual inflammatory reaction to an opportunistic infection in an HIV-positive patient. This syndrome occurs when immunity is restored in the first months of an effective highly active antiretroviral treatment (HAART). First, we described all patients with a cutaneous form of IRIS. Then, between 1992 and 2004 we conducted a retrospective cohort study comparing Herpes Zoster and Herpes Simplex infections among untreated patients, patients treated by HAART for < or = six months, and patients treated for > six months. We observed three cases of atypical leprosy and three original observations: two of these were fistulisation of lymph node histoplasmosis and tuberculosis, the third one reports the recurrence of a treated cutaneous leishmaniasis. Multivariate analysis showed that, after controlling for age, sex and CD4 counts, patients receiving HAART for < or = six months were more likely to develop Herpes Zoster or herpes simplex infections (p < 0.005). Herpes Simplex and Herpes Zoster infections are the two most frequent dermatological manifestations in our tropical setting. Although mycobacterial infections are more rarely observed than in visceral IRIS, the increased incidence of leprosy may be quite significant when the availability of HAART spreads to developing countries. PMID- 16267959 TI - [Evaluation of the therapeutic efficacy of amodiaquine versus chloroquine in the treatment of uncomplicated malaria in Abie, Cote-d'Ivoire]. AB - The WHO 14-days' test and an in vitro survey were carried out to study the efficacy of amodiaquine versus chloroquine in Abie, a hyperendemic village in the southern forest area of Cote-d'Ivoire. One hundred and nineteen children less than 15 years old suffering from uncomplicated malaria were randomised. Among these, 62 were given amodiaquine treatment and 57 chloroquine treatment. both 4 aminoquinoleines were administered at the same dose of 30 mg/kg spread over three days by 10 mg/kg/day. Before the drug was administered, parasites were taken from some patients of each group and were evaluated in vitro to both drugs. In vivo, the amodiaquine treatment shows 95% of clinical success, 2% of early clinical failures and 3% of late clinical failures. For the chloroquine treatment, the rates are respectively. 79%, 7% and 14%. However, some patients still had a level of parasitaemia for both treatments but were asymptomatic. These parasites were found to be resistant in vitro. The authors recommend that the treatment to be used in Abie must be firstly amodiaquine followed by sulfadoxine-pyrimethamine in cases where there is persistent asymptomatic parasitemia. PMID- 16267960 TI - [The incidence of snakebite in a rural zone of southeastern Senegal]. AB - In order to complete an exploratory study on the risk of death due to snakebite in a rural zone of South-Eastern Senegal, we have carried out a survey to estimate the incidence of snakebites in the same population. The study made on a sample of almost 600 subjects showed an annual incidence of 677 bites per 100.000 inhabitants, that is one of the most important rate ever reported in the world until now. Based on these results and data collected previously on deaths due to snakebites in this same population, we provide an estimate of snakebite case fatality rate of 2.1% in this area of Senegal. PMID- 16267961 TI - [Effect of pyrethoid and organophosphate bi-treated nets on the behaviour of Kdr+ Anopheles gambiae mosquitoes]. AB - The efficacy of mosquito nets treated with bifenthrin (pyrethroid) and chlorpyrifos-methyl (organophosphate), alone and in mixture, was evaluated against the pyrethroid-resistant strain (VKPR) of Anopheles gambiae using the tunnel test technique. Results first showed that bifenthrin and chlorpyrifos methyl kept a good excito-repellency effect against resistant mosquitoes despite the low doses applied on nets. With the mixture, irritancy significantly increased, which consequently limited the duration of exposure of females to impregnated surface. This phenomenon most probably explained the absence of synergism between bifenthrin and chlorpyrifos-methyl concerning the insecticidal activity of this mixture. This study demonstrated the importance of the mosquito behaviour in the global effectiveness of insecticide treated materials and in the development of alternative strategies for vector control. Concerning the prospective insecticide mixtures, not only will we have to consider the insecticidal properties of each insecticide selected, but also their excito repellency which affects the time of duration of mosquitoes on the treated surface. PMID- 16267962 TI - Laboratory diagnosis of Ebola and Marburg hemorrhagic fever. AB - The control of Filovirus outbreaks can be greatly enhanced by timely laboratory confirmation of infection or the identification of alternative disease processes. The status of current laboratory diagnostics for Ebola and Marburg virus infections is discussed in terms of the assays available and their interpretation. In addition, the role of field-based laboratory support and its limitations and capabilities in an outbreak response setting, especially in regards to real-time PCR and immunofiltration assays, is presented. PMID- 16267963 TI - Ebola virus circulation in Africa: a balance between clinical expression and epidemiological silence. AB - Nearly thirty years after the first epidemics, Ebola virus (EBOV) remains hardly described, its transmission unclear and its reservoir elusive. Soon after the Ebola fever outbreak and virus discovery in 1976 and in order to investigate the distribution of EBOV in Central Africa, several countries including a range of ecological zones were investigated in the early 1980s, using extensive survey: Central African Republic (CAR), Cameroon, Chad, Congo, Gabon and Equatorial Guinea. Since 1992, ELISA antibody test along with a RT-PCR have been used to detect specific virus antibodies and characterize viral RNA. The widely separated geographic locations of outbreaks have suggested that the reservoir and the transmission cycle of EBOV are probably closely associated with the rain forest ecosystem, what is supported by the distribution of antibodies. The fact that outbreaks seldom occur suggests the presence of a rare or ecologically isolated animal reservoir having few contacts with humans and non-human primates. However various serological investigations showed a high prevalence in humans without any pathology reported. This suggests a circulation of both pathogenic and non pathogenic strains as well as more frequent contacts with man than expected, and could partially explain fifteen years of Ebola fever silence between the emergence and re-emergence of Ebola virus in the Congolese basin. Nowadays, largely enlightened by the study of recent epizootic and epidemic manifestations of EBOV in Gabon and neighboring countries, EBOV natural history starts to be understood as for the fundamentals of epizootic in non-human primates and chains of transmission. PMID- 16267964 TI - [Clinical management of patients and deceased during the Ebola outbreak from October to December 2003 in Republic of Congo]. AB - Outbreaks of Ebola virus hemorrhagic fever (EVHF) have been reported since 2001 in the Cuvette Ouest department, a forested area located in the Western North of Congo. At the end of October 2003 a new alarm came from this department which was quickly confirmed as being an epidemic of EVHF. The outbreak response was organized by the ministry of health with the assistance of an international team under the aegis of WHO. The case management of suspect cases was done in an isolation ward set up at the hospital; when patients refused to go to the ward for care they were isolated in their house according to a protocol "transmission risks reduction at home". Safe burials were performed by specialized teams which respected the major aspects of the funeral to allow the process of mourning of the families. An active surveillance system was set up in order to organize the detection of new cases and the follow-up of their contacts. A case definition was adopted. From October 11 to December 2, 2003, 35 cases including 29 deaths were reported, 16 cases were laboratory confirmed. The first four cases had been exposed to monkey meat (Cercopithecus nictitans). The epidemic spread was due to family transmission. The population interpretation of the disease, in particular questions around wizards and evil-minded persons, is a factor which must be taken into account by the medical teams during communication meetings for behavioral change of the populations. The case management of patient in isolation wards to prevent the transmission of the virus in the community remains the most effective means to dam up Ebola virus hemorrhagic fever outbreaks. The good perception by the community of the safe funerary procedures is an important aspect in the establishment of confidence relations with the local population. PMID- 16267965 TI - [Multiple Ebola virus haemorrhagic fever outbreaks in Gabon, from October 2001 to April 2002]. AB - Outbreaks of Ebola virus haemorrhagic fever have been reported from 1994 to 1996 in the province of Ogooue Ivindo, a forest zone situated in the Northeast of Gabon. Each time, the great primates had been identified as the initial source of human infection. End of November 2001 a new alert came from this province, rapidly confirmed as a EVHV outbreak. The response was given by the Ministry of Health with the help of an international team under the aegis of WHO. An active monitoring system was implemented in the three districts hit by the epidemic (Zadie, Ivindo and Mpassa) to organize the detection of cases and their follow up. A case definition has been set up, the suspected cases were isolated at hospital, at home or in lazarets and serological tests were performed. These tests consisted of the detection of antigen or specific IgG and the RT-PCR. A classification of cases was made according to the results of biological tests, clinical and epidemiological data. The contact subjects were kept watch over for 21 days. 65 cases were recorded among which 53 deaths. The first human case, a hunter died on the 28th of October 2001. The epidemic spreads over through family transmission and nosocomial contamination. Four distinct primary foci have been identified together with an isolated case situated in the South East of Gabon, 580 km away from the epicenter. Deaths happened within a delay of 6 days. The last death has been recorded on the 22nd of March 2002 and the end of the outbreak was declared on the 6th of May 2002. The epidemic spreads over the Gabon just next. Unexplained deaths of animals had been mentionned in the nearby forests as soon as August 2001: great primates and cephalophus. Samples taken from their carcasses confirmed a concomitant animal epidemic. PMID- 16267966 TI - Medical anthropology and Ebola in Congo: cultural models and humanistic care. AB - Seldom have medical anthropologists been involved in efforts to control high mortality diseases such as Ebola hemorrhagic fever (EHF) This paper describes the results of two distinct but complementary interventions during the first phases of an outbreak in the Republic of Congo in 2003. The first approach emphasized understanding local peoples cultural models and political-economic explanations for the disease while the second approach focused on providing more humanitarian care of patients by identifying and incorporating local beliefs and practices into patient care and response efforts. PMID- 16267967 TI - Ebola and great apes in Central Africa: current status and future needs. PMID- 16267968 TI - Climate-based health monitoring systems for eco-climatic conditions associated with infectious diseases. AB - Despite a century of confidence and optimism in modern medicine and technology inspired by their often successful prevention and control efforts, infectious diseases remain an omnipresent, conspicuous major challenge to public health. Effective detection and control of infectious diseases require predictive and proactive efficient methods that provide early warning of an epidemic activity. Of particular relevance to these efforts is linking information at the landscape and coarser scales to data at the scale of the epidemic activity. In recent years, landscape epidemiology has used satellite remote sensing and geographic information systems as the technology capable of providing, from local to global scales, spatial and temporal climatic patterns that may influence the intensity of a vector-borne disease and predicts risk conditions associated with an epidemic. This article provides a condensed, and selective look at classical material and recent research about remote sensing and GIS (geographic information system) applications in public health. PMID- 16267970 TI - [Doing the homework]. PMID- 16267969 TI - [Training the trainers seminar and analysis of the Ebola virus hemorrhagic fever outbreaks in central Africa from 2001 to 2004. (Brazzaville, Republic of Congo, April 6-8, 2004]. PMID- 16267971 TI - [Balsam for skin and spirit--a project in health promotion: rediscovering personal resources]. PMID- 16267972 TI - [Hygiene in nursing--4: Preventing wound infections]. PMID- 16267973 TI - [Rheumatologic nursing care: retaining independence as long as possible]. PMID- 16267974 TI - [Ergotherapy in rheumatic diseases: managing the daily routine]. PMID- 16267975 TI - [Joint operation as ultima ratio in rheumatic arthritis? Retaining function is the goal]. PMID- 16267976 TI - [Nursing in juvenile idiopathic arthritis: not making the illness the center of care]. PMID- 16267977 TI - [Patient education in systemic vasculitis: maintaining quality of life]. PMID- 16267978 TI - [Nursing consultations for chronic patients: help for self-care]. PMID- 16267979 TI - [Principles of patient-oriented information and counseling: using pedagogic strategies appropriate for the situation]. PMID- 16267980 TI - [Counseling in nursing: establishing basic principles of education]. PMID- 16267981 TI - [Idiolectic approach for counseling in nursing: understanding other people]. PMID- 16267982 TI - [Introducing change actively and with goal orientation: counselors have a key role]. PMID- 16267983 TI - [Continuing education for palliative care nursing: giving support at the end of life]. PMID- 16267984 TI - [Nurse counseling on anticipated will declaration: analysis of personal values and wishes]. PMID- 16267985 TI - [From case to case: (no) reason for despair]. PMID- 16267986 TI - [Fever in critically ill patients: frequent causes, diagnostic procedures, and therapy]. AB - Fever is an adaptive physiological process. In reaction to specific proteins (pyrogens) distributed by the blood the reference temperature increases, thus the organism has to produce heat. In intensive care units approximately 50 percent of fever episodes are caused by hospital acquired infections, while other episodes result from non-infectious diseases or their aetiology remain undetected. A special kind of fever is the hyperthermia following malfunctioning hypothalamic regulation after central nervous diseases or injuries. In patients with severe head injuries any increase of temperature has to be prevented as hyperthermia is associated with more adverse outcomes. If the hyperthermia can be attributed to other aetiological factors, a decrease of temperature is not desirable provided that the consequences of fever do not jeopardize the patient for further health risks. As a basis of therapy, the aetiology of fever episodes should be diagnosed taking recent scientific knowledge and guidelines into consideration. The most reliable temperature values are measured by using invasive sensors, particularly in pulmonary arterial catheters or bladder catheters. Alternatively usual thermometers can be applied, due to unreliable results excepting in the axilla. PMID- 16267988 TI - Proposed Medicare fee schedule includes 4.3 percent cut, policy changes. PMID- 16267987 TI - Ready or not, pay for performance is here. PMID- 16267989 TI - Pulling their own weight. PMID- 16267990 TI - Mississippi practice grapples with Katrina. PMID- 16267991 TI - Managing the difficult doc: three case examples. PMID- 16267992 TI - Payer contracts: do you know if you're being reimbursed appropriately? PMID- 16267993 TI - Off to a slow start... AB - Challenging common estimates of electronic health record (EHR) adoption among medical group practices, a new comprehensive study by the Medical Group Management Association (MGMA) and the University of Minnesota Division of Health Services Research and Policy has found that just 14.1 percent of medical group practices have fully implemented EHR. While the current adoption rate is low, the study also found that growth in EHR adoption over the next two years could be dramatic. Learn the technology outlook for medical groups. PMID- 16267994 TI - For love of the profession. Benjamin S. Snyder--Harry J. Harwick Lifetime Achievement Award. PMID- 16267995 TI - Evaluating freestanding ambulatory surgery center ventures. AB - Projecting revenues and expenses The second of a two-part series, this article discusses the projected revenues and expenses of a hypothetical ambulatory surgery center and analyzes the potential impact of a hypothetical new payment method by the Centers for Medicare & Medicaid Services. PMID- 16267996 TI - Kickin' it. PMID- 16267997 TI - A primer for nursing students on current issues in risk management. PMID- 16267998 TI - School of nursing becomes University's first flagship program. PMID- 16267999 TI - FIU school of nursing confers distinguished service medallion on Thelma Gibson. PMID- 16268000 TI - University of Central Florida College of Health and Public Affairs School of Nursing. PMID- 16268001 TI - FONE survey on nursing research resources. PMID- 16268002 TI - Florida Center for Nursing. PMID- 16268003 TI - A study of a structured exercise program with members of an ICCD Certified Clubhouse: program design, benefits, and implications for feasibility. AB - Individuals with serious mental illness (SMI) have significantly greater risk of comorbid health problems and premature death, and there is need for interventions that can improve physical fitness and overall health. Accordingly, a study was conducted which evaluated the effectiveness of a structured physical exercise program that was developed as part of a wellness project in an ICCD Certified Clubhouse. Seventeen clubhouse members completed a 16-week program with evidence of significant improvement in aerobic capacity and perceived mental health as well as positive trends in perceived improvements in physical and social functioning. Qualitative data indicated satisfaction with the program by all participants, especially the value of group support, while also highlighting the need for greater attention to nutrition as part of a future program. Moreover, the study found that a structured exercise program can be successfully provided to members of an ICCD Certified Clubhouse. PMID- 16268004 TI - A narrative approach to the evaluation of supportive housing: stories of homeless people who have experienced serious mental illness. AB - We present the findings of a narrative approach to the evaluation of supportive housing for formerly homeless people who have experienced serious mental illness. According to the accounts of 11 men and 9 women, their youth and adult years were filled with personal problems, troubled relationships, and a lack of adequate social resources. Since entering supportive housing, participants noted more stability in their lives and the beginning of journeys to recover positive personal identities, restore or develop new supportive relationships, and reclaim resources vital to leading lives with dignity and meaning. The findings add to the literature on housing interventions for this population in suggesting many positive gains beyond reductions in homelessness and hospitalization. PMID- 16268005 TI - Work-related self-efficacy among community residents with psychiatric disabilities. AB - A new measure of work-related self-efficacy for people with psychiatric disabilities is reported. The 37-item scale measures self-efficacy in four relevant activity domains: (1) vocational service access and career planning, (2) job acquisition, (3) work-related social skills, and (4) general work skills. The scale was developed in a 12-month longitudinal survey of urban residents diagnosed with schizophrenia or schizoaffective disorder (n=104). Results indicate validity of both a four-factor structure differentiating four core skill domains, and a single factor representing total work-related self-efficacy. The favorable psychometric properties support further research and trial applications in supported employment and psychiatric vocational rehabilitation. PMID- 16268006 TI - Predictors of psychiatric hospitalization for adults with co-occurring substance and mental disorders as compared to adults with mental illness only. AB - Predictors of psychiatric hospitalization, predisposing, enabling and need, of adults with co-occurring mental and substance disorders were compared to predictors for adults with a mental illness only. Research participants were 1613 users of crisis intervention services. Findings using Cox regression show that dually-diagnosed individuals were more likely to be hospitalized. Enabling and need factors were important predictors for both groups. Disruptive behavior was a predictor for dually-diagnosed clients but not for clients with mental illness only. Findings suggest that outpatient mental health services are less well equipped to address a psychiatric crisis when it was accompanied by substance use issues. PMID- 16268007 TI - The role of a fitness intervention on people with serious psychiatric disabilities. AB - The purpose of this study was to determine the effects a health education and exercise program would have in limiting weight gain and in improving fitness and psychological parameters in adults with mood or psychotic disorders. Thirty volunteers were randomly assigned to the healthy lifestyle group (HL) or a control group. The HL group engaged in exercise for 12 weeks. Pre- and post exercise testing was conducted to assess body fat, lipid profile, and cardiovascular fitness. Educational seminars were held weekly. The intervention group evidenced greater weight loss than the control group, although not statistically significant. Significant differences were observed in ratings of general health (p < .05) and empowerment (p < .01). Trends suggest that exercise interventions may encourage weight loss, particularly if barriers to full participation can be addressed. Additionally, such interventions may contribute to "perceived" well-being even among those with subclinical participation. PMID- 16268008 TI - The job developer's presence in the job interview: is it helpful or harmful to persons with psychiatric disabilities seeking employment? AB - This study examines the effect of having an employment specialist join supported employment participants at job interviews. The sample consists of 45 individuals with psychiatric disabilities who sought job placement services during a 28-month period between December 1991 and March 1994. Seventy-nine percent of the job offers occurred when the job developer was present in the interview as compared to only 21% when the job applicants went alone. The finding challenges "current" conventional wisdom about having rehabilitation staff at the worksite. Rehabilitation professionals need to take a fresh look at program policies and practices regarding on-site services to SE participants. PMID- 16268009 TI - Relationships between patient, family, and significant other disposition preferences in psychiatric emergency services and the clinical symptom ratings and disposition decisions of psychiatrists. AB - A previous paper examined the relationship between 10 clinical ratings made by psychiatrists and 465 psychiatric emergency service (PES) dispositions. A logistic regression model explained 52% of the variance in the psychiatrist's decisions to admit to inpatient services or release. The 5 clinical ratings in the model, rated on 8-point scales, were severity of danger to self, psychosis, inability to care for self, impulse control, and depression. The current paper examines the relationships between patient and family/significant other dispositional preferences expressed in the PES, and the psychiatrist's disposition decision and the 10 clinical ratings. It found that the dispositional preferences of the patient, family, and other individuals (community clinician, police, or ambulance driver) were very consistent with the actual decision made by the PES psychiatrist. Further, it found that these preferences explained an additional 35% of the variability in the psychiatrist's decision to admit or release. Finally, the ten clinical ratings made by the PES psychiatrist explained only a small amount of the variance in the dispositional preferences, and some of the significant coefficients were in the opposite direction expected. Patient, family and other significant individuals' expressed preferences and expectations in the PES are important and should be routinely recorded in the charts. The small relationships between clinical ratings and preferences suggests that the reasons the family/significant other and the patient seeks hospital admission or release may differ with psychiatrists' reasons for disposition. PMID- 16268010 TI - A program of supported education for adult Israeli students with schizophrenia. AB - This report describes a supported education program (SEP), a novel community based program of rehabilitation for psychiatric patients, started in Israel in 1999. Objectives, target population and activities are described. In addition, initial findings of two program-related studies are presented. One study explored the relationship of psychological distress with students' coping strategies and perceived social support. The second study is a survey of mental health literacy and attitudes of heads of schools for adult education where the SEP was implemented. Studies' findings suggested a "case for action" with the following objectives: to develop interventions to prevent distress among SEP users, and to enhance mental health knowledge and foster positive attitudes among school personnel aimed at decreasing dropout rates and encourage the utilization of the opportunities provided by SEP. PMID- 16268011 TI - How the pace program builds a recovery-transformed system: results from a national survey. PMID- 16268013 TI - E-commerce stats refine search for supply savings. PMID- 16268012 TI - Acceptance and change: the dialectic of recovery. AB - While a good deal of research has been conducted on the factors associated with recovery, we still need a clearer understanding of the dialectic between behavior and attitude. This research uses semi-structured interviews with two groups of mental health consumers, one receiving more intensive ACT services and the other receiving less intensive case management services, to ascertain the illness management strategies used by those who have been transferred to less intensive services. Our research suggests that while engaging in community, vocational, and church activities are important, those who are further along the road to recovery have gained an understanding and acceptance of their illness that allow them to use these activities as effective illness management strategies. PMID- 16268014 TI - Wound care prices set to rise, survey says. PMID- 16268015 TI - Rethinking disability and what to do about it: disability studies and its implications for occupational therapy. AB - Disability studies seeks to reframe rehabilitation's understanding and responses to disability. Disability scholars point out that rehabilitation's perspectives and practices are not objective, but instead reflect particular historical and ideological forces. By demonstrating how rehabilitation practitioners can unintentionally do things that are unhelpful or even harmful, disability scholars challenge the profession of occupational therapy to reconsider aspects of practice, education, and research. In order to provide a context for a special issue devoted to disability studies, this paper examines disability studies' major critiques of rehabilitation and considers their implications for occupational therapy. The paper identifies ways that occupational therapy can continue to respond reflectively to the themes of disability studies. It also identifies a number of questions raised by disability studies that will need to be addressed in practice, education, and research. PMID- 16268016 TI - Can the social model explain all of disability experience? Perspectives of persons with chronic fatigue syndrome. AB - OBJECTIVE: The social model of disability has had a major influence on the academic field of disability studies and on contemporary understandings of the causes and experience of disability. The purpose of this study was to examine the adequacy of the social model for explaining the disability experience of persons with chronic fatigue syndrome (CFS). METHODS: This qualitative study examined the experiences of 47 adults with CFS participating in a research project that aimed to evaluate a participant-designed rehabilitation program. Data were aggregated from focus group interviews, open-ended questionnaires, progress notes, and from a program evaluation questionnaire. Data analysis was based on a grounded theory approach and used triangulation of multiple data sources and member checks to assure dependability of findings. RESULTS: Four themes emerged from the analysis: (1) minimization and mistrust of the disability; (2) negative experiences of impairment; (3) lack of identification with the disability community; and (4) the focus on advocacy as a quest for legitimacy. These themes varied in the extent to which they conformed to the principles set forth by the social model. CONCLUSIONS: Although the social model has important contributions to lend to occupational therapy practice, it is important to recognize that it may not capture the full reality of disability. In particular, the social model has serious limitations in describing the disability experience of individuals with disabilities who do not have visibly obvious disabilities and whose impairments do not conform to the traditional viewpoint of disability. PMID- 16268017 TI - Habitus and the embodiment of disability through lifestyle. AB - One aim of disability studies is to accurately portray the experience of persons who live with disabilities. Often this experience generates knowledge that challenges institutional and attitudinal stereotypes. In this study, disability is conceptualized as a measurable space and place characterized by objective differences among groups. Bourdieu's concept of habitus is proposed as a way to understand how a social space is created and maintained by everyday life experience. Linking Bourdieu's understanding of social space to the sociology of health lifestyles, the paper further proposes that classes of individuals enact individual and collective behavior that respond to life choice and life chance. Using measures from the Craig Handicap Assessment and Reporting Technique clustered to reveal patterns of community participation 2 years after spinal cord injury, the study offers an alternative way for modeling knowledge of disability as embodied categories of action expressed as differentiated health lifestyles. PMID- 16268018 TI - Disability studies at the population level: issues of health service utilization. AB - OBJECTIVE: In this paper, I propose a population-level analysis of disability to raise issues of access and equity in terms of use of health services. METHOD: The study was a cross-sectional analysis of the National Population Health Survey (Statistics Canada, 1998-1999). The sample consisted of 10,898 adults between 20 and 64 years of age. FINDINGS: The study showed that adults with disabilities used significantly more of all types of health professionals and health services than nondisabled adults. Disability was a significant determinant of all types of health service use, representing a two- to threefold increase in risk of seeing health professionals. Although poor health explained a large proportion of variance attributable to use of medical and nursing services, it did not explain the use of other allied health services. The results are interpreted in terms of structural barriers to access to health services. The findings also remind us of the potential role for occupational therapists as advocates within the health care system for persons with disabilities. PMID- 16268019 TI - The problem with prevention: the case of spina bifida. AB - In this paper, I present a viewpoint about prevention and spina bifida that is not usually expressed within the occupational therapy literature. Using an autoethnographic account, I convey my experiences as a person with impairments from spina bifida in order to problematize current preventive efforts undertaken to eradicate this birth defect. This self-reflexive account connects my personal experiences to historical and medical views about spina bifida. The messages inherent in preventive efforts are discussed from a disability rights perspective. Occupational therapists are challenged to examine their attitudes toward disability, act as advocates in their practice, and, in a more informed manner, support or contest policy initiatives. PMID- 16268020 TI - Resident and staff perceptions of barriers to independence and employment in supportive living settings for persons with AIDS. AB - The purpose of this study was to identify perceived barriers to independent community living and employment among clients and staff members in a transitional living facility for persons with AIDS. This qualitative study used focus groups to collect these perceptions from staff members (N = 21) and clients (N = 16). Whereas staff identified both systemic and personal barriers, clients only identified systemic barriers. These findings suggest that both clients and staff recognize the types of environmental barriers to participation that have been identified by disability scholars. The findings also underscore a consequential gap between clients' exclusive emphasis on environmental barriers and staff emphasis on clients' personal barriers that impact upon participation. PMID- 16268021 TI - Integrating disability studies concepts into occupational therapy education using service learning. AB - This article describes an occupational therapy educational program's experience with service-learning courses that has fostered student learning about service to the community and disability as a multidimensional construct. Faculty-reflective perspectives about disability and ways to enhance learning about disability as a human experience are presented as an important consideration for health care education curriculum design and course development. Through review of educational evaluation described in research on service learning, the authors used a multi method assessment matrix to capture students' perspectives on their service learning. The community project investigated accessibility issues that persons with disabilities encountered in social participation at community arts venues. Results from student surveys, interviews, focus groups, and journal entries indicated that service learning contributed to occupational therapy students' appreciation and understanding about disability as an individual, environmental, and societal construct. PMID- 16268022 TI - Introducing disability studies to occupational therapy students. AB - This article is a work of collaborative ethnography about teaching and learning disability studies within the context of an occupational therapy graduate program. In spring 2004,14 occupational therapy students were introduced to disability studies by their cultural anthropologist (nonoccupational therapist) course instructor. During the one-credit course, they were expected to complete readings, watch films, attend guest lectures, and make a site visit. The occupational therapy students were required to write a journal to record personal reactions and new insights gained from these experiences. This article focuses on a thematic analysis of the students' journaled responses to the film "Dance Me to My Song," and a site visit to a local Independent Living Center. Students were expected to analyze these experiences from both disability studies and occupational therapy perspectives. The article addresses philosophical and practical differences between occupational therapy and disability studies and identifies opportunities for collaboration between occupational therapists and independent living specialists. PMID- 16268023 TI - The Olmstead decision: landmark opportunity or platform for rhetoric? Our collective responsibility for full community participation. AB - Occupational therapy's core values and founding beliefs reflect a commitment to the attainment of a self-directed life that enables full community participation. The reality that this desired outcome remains unattainable for millions of persons with disabilities due to the institutional bias of current public policies cannot be ignored. The Supreme Court's landmark Olmstead decision provides a judicial mechanism to right this fundamental injustice. The executive branch's response to the Olmstead decision, the New Freedom Initiative (NFI), offers the potential to finally end the societal segregation of persons with disabilities. Occupational therapy practitioners share a moral collective responsibility to advocate for the major systemic changes that will be needed to fully implement the NFI and achieve the promise of the Olmstead decision. By joining with the disability rights movement, occupational therapy practitioners can become effective activists to help persons with disabilities attain full participation in life. PMID- 16268024 TI - An occupational perspective on the concept of participation in the International Classification of Functioning, Disability and Health--some critical remarks. AB - The International Classification of Functioning, Disability and Health (ICF; World Health Organization, 2001) provides an international and interprofessional scientific basis for understanding and studying health. The concept of participation plays an important role in the classification and has become a central construct in health care, rehabilitation, and in occupational therapy. The aim of this paper is to provide a critical analysis of the concept of participation in the ICF. As background, the origins and current presentation of the ICF are presented. The use and function of the ICF and the contemporary discussions regarding the classification are reviewed. An occupational perspective on participation in the ICF reveals major shortcomings regarding the subjective experience of meaning and autonomy. Furthermore, the ICF has limitations in capturing different kinds of participation in a single life situation. Following these analyses we discuss the advantages and shortcomings of using the ICF, and how an occupational perspective can contribute to an ongoing discussion about the development of the ICF. PMID- 16268025 TI - Nothing about us without us: searching for the narrative of disability. PMID- 16268026 TI - Therapists I have known and (mostly) loved. AB - This manuscript embeds the author's firsthand experiences with pediatric occupational therapy with her expertise as a developmental psychologist interested in families and their collaborations with service professionals. The author's own experiences are used to illustrate points made in literature regarding parent-professional collaborations, as well as points made in interviews of other parents of young children with special needs. It is hoped that the article can serve as a "starting off" point for discussions and partnership building between pediatric occupational therapists and the children and families they serve. PMID- 16268028 TI - Beyond the horizon: health care for everyone. PMID- 16268027 TI - Building a professional tapestry. PMID- 16268029 TI - The need for mentorship. PMID- 16268030 TI - 1000 CNA certified nephrology nurses by 2006: is it possible? PMID- 16268031 TI - Smoking cessation treatment options. PMID- 16268033 TI - The role of peer mentorship in job satisfaction of registered nurses in the hemodialysis unit. AB - The effects of health care economic constraints and restructuring have had a deleterious effect on the job satisfaction of registered nurses. Canada is also experiencing an aging nursing workforce, and a nursing shortage due, in part, to stressful work environments. Concurrently, workloads are increasing as nurses in dialysis units care for older patients with many co-morbid conditions. Mentorship has been shown to positively impact workplace culture and job satisfaction for both the mentor and the individual being mentored. The role of peer mentorship within the context of a community-based hemodialysis unit is explored as a potential strategy to improve job satisfaction of dialysis nurses. PMID- 16268032 TI - The evolution of a mentoring relationship. AB - My involvement in a formalized mentoring partnership evolved from an advanced clinical practice fellowship (ACPF) through the Registered Nurses Association of Ontario (RNAO). The RNAO provides opportunities for registered nurses across the province to learn more about their special interest by offering fellowships for those who are committed to expanding their knowledge while meeting their desired goals and objectives. My experience in this project far exceeded my expectations. Not only did I meet my goal on a professional level, I also learned the value of mentoring and the impact that such an experience can have on a person overall. This paper discusses how my mentor guided me through my fellowship and how our relationship typified the mentor-protege role as described in several articles. PMID- 16268035 TI - Mentoring: in the eye of the beholder. PMID- 16268036 TI - An interview with CANNT Western Region VP--Rick Luscombe. Interview by Pady Dunn. PMID- 16268034 TI - Resources for preceptors. PMID- 16268037 TI - The interrelationships among thinking skills, research knowledge, and evidence based practice. PMID- 16268038 TI - Views on the nursing faculty shortage. PMID- 16268039 TI - Views on the nursing faculty shortage. PMID- 16268040 TI - Teaching thinking and reaching the limits of memorization: enacting new pedagogies. AB - Contemporary practice environments are complex and require students and new graduates to possess high-level thinking capabilities to ensure patient safety and quality of care. To achieve this goal, many nurse educators are exploring new pedagogies (critical, feminist, phenomenological, and postmodern) to teach students the practices of thinking that contemporary clinical situations demand. These new pedagogies offer alternatives to the predominant use of conventional pedagogies, which emphasize students' memorization of content and its subsequent application in practice situations. This study, using Heideggerian hermeneutics, documents how teachers are using new pedagogies in ways that shift their attention to teaching thinking and away from devising strategies to help students memorize and apply more content. One of the themes that emerged from this study, Teaching Thinking and Reaching the Limits of Memorization, and two subthemes, Thinking AS Memorization and Recall, and Beyond Memorization: Thinking in Context are presented. PMID- 16268041 TI - Predictors of novice nurses' use of intuition to guide patient care decisions. AB - Nurses routinely report using intuition to guide decisions about patient care, although they use it covertly because of difficulty explaining the sources of their intuitions to colleagues. To help nurse educators guide students toward open discussion and appropriate use of intuition, this study compared personal, interpersonal, and professional experiences suggested by past research for their association with the use of intuition by nurses. A questionnaire completed by 323 novice nurses measured use of intuition with an 18-item subscale identified by Miller from the Miller Intuitiveness Instrument. Measures of personal experience included age, gender, hospitalizations, self-esteem, and religiosity. Interpersonal experience included parenthood and social support from family and friends. Professional experience included grade point average and months of on the-job experience. Multiple regression revealed that novice nurses use intuition more to guide patient care if they are older, have had more hospitalizations, and experience more social support. PMID- 16268042 TI - The effect of problem-based learning on nursing students' perceptions of empowerment. AB - This study tested Kanter's structural empowerment theory within a university nursing student population. Differences in perceptions of empowerment among nursing students enrolled in either a problem-based learning (PBL) or a conventional lecture learning (CLL) program were examined, as well as the relationship between perceptions of structural empowerment in the learning environment and feelings of psychological empowerment. Participants completed measures of structural and psychological empowerment adapted to educational settings, as well as measures related to exposure to various learning strategies in their programs and clinical problem-solving abilities. Students in the PBL program (n = 41) had significantly higher perceptions of structural and psychological empowerment than students in the CLL program (n = 67). Regardless of academic program, structural empowerment was strongly positively related to psychological empowerment. The results of this study are the first to support the applicability of Kanter's theory to nursing education settings. PMID- 16268043 TI - Introducing students to research: the road to success. AB - Introduction of the terminology associated with research can overwhelm both students and faculty. In this article, I describe an interactive method for teaching undergraduate nursing students about the concepts involved in the research process. The "Research Road to Success" is a board game created to promote active learning using repetition, peer collaboration, group dynamics, and questioning. The game has been part of the curriculum for 3 years, with positive results. Nurse educators and students can benefit from the incorporation of creative methods to stimulate interest in and understanding of nursing research. PMID- 16268044 TI - Actualizing scholarship in senior baccalaureate nursing students. AB - Nursing scholarship is difficult to define and often equated with either research and publication in academia or academic achievement of students, but the definition has recently been broadened to include clinical scholarship. This article emphasizes the connections between the theory of nursing scholarship and its application to advanced clinical practice for senior baccalaureate nursing students. Strategies used to enhance students' ability to operationalize scholarship include reflective journals, development of an abstract and professional paper, and presentation of a paper or poster at the student-led Nursing Scholarship Forum. The Forum is evaluated as a learning activity that helps students actualize scholarship. The results of the evaluation indicate this type of scholarly activity enhances nursing students' transition into the professional practice role. PMID- 16268045 TI - A service learning research methods course. AB - Incorporation of service learning into an undergraduate nursing research course has the potential to invigorate baccalaureate nursing students' interest in research through real-world connections. In this article, I describe the development and implementation of an undergraduate nursing research course that incorporated service learning. The course experience proved to be enlightening for both the students and instructor, and propelled students to continue their research beyond the course requirements. PMID- 16268046 TI - The prescription pad as a teaching tool. PMID- 16268047 TI - Incorporating CQI concepts into student community health projects. PMID- 16268048 TI - The insulin-like growth factor system: IGFs, IGF-binding proteins and IGFBP proteases. AB - Insulin-like growth factors (IGF-I/-II) are not only the endocrine mediators of growth hormone-induced metabolic and anabolic actions but also polypeptides that act in a paracrine and autocrine manner to regulate cell growth, differentiation, apoptosis and transformation. The IGF system is a complex network comprised of two growth factors (IGF-I and -II), cell surface receptors (IGF-IR and -IIR), six specific high affinity binding proteins (IGFBP-I to IGFBP-6), IGFBP proteases as well as several other IGFBP-interacting molecules, which regulate and propagate IGF actions in several tissues. Besides their broad-spectrum physiological and pathophysiological functions, recent evidence suggests even a link between IGFs and different malignancies. PMID- 16268049 TI - Hypertriglyceridemia, the coronary heart disease risk marker "solved". AB - Both epidemiological and intervention studies have shown that hypertriglyceridemia is a significant cardiovascular risk factor. The large variation of the triglyceride values is explained by the influence of several modifying factors, which are difficult to standardise. Therefore hypertriglyceridemia should be considered rather as risk marker, than risk factor. The measurement of the apolipoprotein CIII level, which is a more stable parameter of the triglyceride rich lipid particles, is now becoming more widespread. This parameter is also able to substitute the assessment of the small dense LDL form that has a controversial significance. The clinical benefit of reduction of triglyceride concentration and the accompanying increase of HDL cholesterol level by fibrates, in the prevention of the coronary heart disease (CHD) events, have been demonstrated in several prospective, placebo-controlled trials. The VA-HIT study, enrolling the largest number of patients, has shown that fibrates have another effect, presumably influencing the insulin resistance independently of lipid levels that is also able to reduce the CHD events. PMID- 16268050 TI - Testosterone and endurance exercise: development of the "exercise-hypogonadal male condition". AB - During the last 30 years a large number of research studies have been conducted examining reproductive endocrine dysfunction in exercising women. The number of similar studies examining men is still relatively small. Nevertheless, an increasing amount of research studies in men indicate endurance exercise training has significant effects upon the major male reproductive hormone, testosterone, and the hypothalamic-pituitary-testicular axis that regulates reproductive hormones. This review article addresses one reproductive endocrine dysfunction found in exercising men, what has been deemed the "exercise-hypogonadal male condition". Specifically, men with this condition exhibit basal (resting-state) free and total testosterone levels that are significantly and persistently reduced. The exact physiological mechanism inducing the reduction of testosterone is currently unclear, but is postulated to be a dysfunction (or perhaps a readjustment) within the hypothalamic-pituitary-testicular regulatory axis. The time course for the development of the "exercise-hypogonadal condition" or the threshold of exercise training necessary to induce the condition remains unresolved. The potential exists for these reduced testosterone levels within the exercise-hypogonadal male to disrupt and be detrimental to some anabolic or androgenic testosterone-dependent physiological processes. Unfortunately, extremely few research studies have addressed whether such processes are affected, and thus findings are inconclusive. Conversely, the alterations in testosterone levels brought about by endurance exercise training have the potential for cardiovascular protective effects and thus could be beneficial to the health of these men. Current evidence suggests this condition is limited to men who have been persistently involved in chronic endurance exercise training for extended periods of time (i.e., years). Many questions, however, regarding the male reproductive endocrine adaptive process to exercise and exercise training remain unanswered, necessitating the need for further research on this topic. PMID- 16268051 TI - Influence of moderate altitude on blood lactate and heart rate in a standardized exercise test in healthy volunteers. AB - Nineteen healthy volunteers were exposed to a standardized exercise test at sea level (SLa), at an altitude of 1700 m before (1700a) and after a moderate 10-day mountain training (1700b), with a final control four weeks later at sea level (SLb). Vital signs, blood lactate and arterial oxygen saturation were determined prior, during or after the exercise test. Whereas systolic blood pressure and heart rate at rest did not change substantially, diastolic blood pressure decreased at the final control (SLb, p<0.05) and oxygen saturation was significantly lower at 1700 m (1700a, 1700b, p<0.01). Lactate at rest increased from 1.16 (SLa) to 1.97 (1700a) mmol/l after acute exposure followed by a slight reduction after adaptation (p<0.05). The mean maximum lactate levels were as follows: 6.03, 10.56, 6.22 and 8.75 (p<0.01). The mean maximum performance increased during the study (225.6, 223.3, 231.6, 248.1 Watt, p<0.01). Lactate versus workload curves did not show a marked shift to the right. No significant changes of maximum heart rates during the exercise test were found. In conclusion, a sojourn at 1700 m provokes an increase of lactate levels with subsequent reduction after acclimatization and has a significant positive impact on the mean maximum performance after moderate mountain training. PMID- 16268052 TI - The contribution of blood chemistry to the electrical resistance of blood: an in vitro model. AB - In order to improve the predictive accuracy of impedance cardiac output, the relationship between blood resistance, chemistry, and hematocrit was examined. Blood samples from sixty-three intensive care (ICU) patients was analyzed for hematocrit, sodium, bicarbonate, urea, total protein, albumin, glucose, and pH, and the electrical resistance of the sample was measured. Multiple regression analysis produced a statistically significant model with resistance as the dependant variable, and the exponent of the hematocrit (Exp[Hct]), pH and blood urea as the independent variables. This study therefore suggests that the accuracy of resistance prediction can be improved by incorporating pH and urea into the resistivity equation. It is to be expected that this in turn will improve the accuracy of impedance cardiac output estimation. PMID- 16268053 TI - Immunohistochemical localization of beta1 and beta4 integrins in mouse endometrium during implantation and early pregnancy. AB - Implantation presents the remarkable synchronisation between the development of embryo and differentiation of endometrium. Cell-cell adhesion is an important phenomenon taking place during blastocyst implantation in uterine membrane. We think that the investigation of existence and the level of integrins in women can be a guide for treatment of infertility. Our purpose in this study was to show expression beta1 and beta4 integrins on gestational days 4, 6, 12 by immunohistochemical methods and to investigate whether beta4 integrin is a useful marker for receptivity. beta1 and beta4 integrin were exhibited on surface epithelium on gestational day 4. On the other hand, strong beta4 immunoreactivity was detected on surface epithelium and glandular cells on gestational day 12 but no beta1 reactivity was present in the surface epithelium and glandular cells on day 12. In conclusion, both beta1 and beta4 integrins may have a role in implantation process because positive immunoreactivity was seen on apical membrane of surface epithelium on day 4 when implantation occurred. The localization to apical pole of surface epithelium suggest a role for beta1, beta4 integrins in initial embryo and endometrium interaction. It does not seem that beta1 integrin has a role supporting pregnancy since expression of beta4 on surface epithelium and glandular epithelium disappeared on day 12. beta4 integrin expression increasing on day 12 of pregnancy leads us to think a possible functional role supporting pregnancy. PMID- 16268054 TI - The effects of chronic aerobic and anaerobic exercises n lymphocyte subgroups. AB - Exercise is the strongest stress to which the body is ever exposed. The body responds to this stress through a set of physiological changes in its metabolic, hormonal and immunological systems. In this study, responses of the immune system to the long-term aerobic and anaerobic exercises have been investigated. Twenty four sedentary male university students and officers participated in this study. Subjects were divided into two groups, each consisting of twelve people. Group-1 (age: 25.67 +/- 3.79 years, height: 174.83 +/- 5.15 cm, body mass: 72.17 +/- 8.05 kg) and Group-2 (age: 24.83 +/- 2.89 years, height: 175.3 +/- 6.68 cm, body mass: 70.67 +/- 6.15 kg). After physical examinations of the two groups, resting ECG, respiratory function tests and metabolic tests with the use of the breath by breath method were completed, and anerobic heart rates at the threshold level were determined. The first group was subjected to exercise using Monark ergometry cycles at a heart rate 10% below the threshold level for 8 weeks, 3 days a week, 30 min a day. The second group exercised at a heart rate 10% above the threshold level for 8 weeks, 3 days a week, 20 min a day. Heart rates were checked with the Polar Test during exercises. Pre-exercise (Ep) venous blood samples were taken from each group before their 1st and 24th exercises. Hb (gr), Hct (%), erythrocyte (x10(6)/microl), leukocyte (x10(6)/microl), leukocyte subpopulations (neutrophil, lymphocyte, monocyte, eosinophil, basophil %) and thrombocyte (x10(6)/microl) values were determined. CD3, CD4, CD8, CD19 and CD56 values were determined by Flow Cytometry method using monoclonal antibodies. The chronic effects of exercise were examined through a comparison of Ep blood samples at the 1st exercise with Ep blood samples at the 24th exercise. While the increase in the total leukocyte number was significant (p<0.05) in the first group, increase in the second group was found to be non-significant. When percentiles of leukocyte subpopulations were taken into consideration, changes in the first and second group were found to be non-significant. When lymphocyte subgroups were examined; in the first group a decrease in CD3 and CD4 percentiles to 7% and 12%, respectively (p<0.05) and a 65% increase (p<0.01) in the CD56 value were observed. In the second group a decrease in CD3 and CD4 percentiles to 13% and 17%, respectively (p<0.05) and a 73% increase (p<0.01) in the CD56 value were observed. The Sample-t Test and The Wilcoxon Test were used for statistical analysis. PMID- 16268055 TI - Investigation of serum leptin levels and VO2max value in trained young male athletes and healthy males. AB - The present study aimed at investigating serum leptin levels of elite young male athletes who have been regularly exercising for a long period of time and males who do not exercise. The study included 24 trained young male athletes and 22 healthy sedentary male subjects. Athletes who participated in the study were from different sports branches and have been regularly exercising for at least 2 years. Serum leptin levels were determined by RIA. VO2max levels were identified during maximal exercise. Lactic acid levels were identified one minute before and one minute after exercise from the fingertip by Pro-lactate kit. As a result of the tests, although BMI values of trained young male athletes and healthy males were close to each other, leptin levels were significantly lower (p<0.01), VO2max values were significantly higher (p<0.01) and test periods were significantly longer (p<0.001) in the former. In conclusion, regular exercise, by reducing body fat percentage, suppresses serum leptin levels. PMID- 16268057 TI - [Pathological aspects of the prolonged forms of the hepatitis "A"]. AB - Prolonged forms of the hepatitis "A" have been examined during the three year multicentric prospected study. AIMS: Of the study from patohistological aspect was: 1) Determinate stage and type of liver damage during the differentiate phase of prolonged HAV infection; 2) The resultats of the researches compared with duration of persistance activity of serum ALT, circulating immune complexes (CIC) and persistence length IgM anti-HAV. MATERIAL AND METHODS: In 21 patients with prolonged Hepatitis "A" have been done Biopsy of Liver with Menhginy needle by intercostal approach. We put the material into formalin and it was processed by standard procedure. Biopsy of liver was preformed between 50-60 days, 60-180 days, 180-210 days of illness. Anti-HAV IgM was determined by ELISE method. CIC have determined photometrically in the poliethylenglycole sediment. RESULTS: of the researches showed that in 21 patients with biopsy Hepatitis acuta was faund in 20%, 55 % residue posthepatitis, 10% patients had nonspecifical finding, 10 % reported hronical persistence hepatitis and 5% chronical active hepatitis. CONCLUSIONS: Authors conclused that during the prolonged HAV infection, all stages of liver damage can be find from acute to chronic illness. There is positive correlation between persistance of maintaince of CiC and IgM anti-HAV in patients sera and staging of patohistological lesions in liver. There is no correlation between activity of CIC, activity ALT in sera and study of liver lesions. PMID- 16268056 TI - The history of bronchial asthma from the Renaissance till the beginning of the twentieth century. AB - The aim of the paper is to give an overview of the history of knowledge on asthma from the Renaissance till the beginning of the 20th century. During this period the clinical picture of bronchial asthma and some etiological factors--like familiarity of the disease, the role of the pollen, psychological factors were cleared. The disease was relatively rare in these periods--the epidemiological explosion came only in the second half of the 20th century. Data on pediatric asthma, before all in Hungary are demonstrated mostly based on the works of Schoepf and Bokai senior in the first half of the 19th century. PMID- 16268058 TI - Vineyard pesticide induced changes in the lungs: experimental studying on rabbits. AB - Our intention was to analyze changes in the lungs in rabbits induced by the inhalation of the Bordeaux Mixture aerosol and determination of the time necessary for development of changes that cause respiratory failure and eventually precancerous changes. To experimental rabbits aerosol was administred for 4 months. Lungs were examined pathohistologically and histochemically with rubeanic acid for copper detection. After 4 months of everyday inhalation lung tissue showed diffuse inflammation in all experimental animals, but without granulome formation and fibrosis. The bronchial epithelia showed basal hyperplasia and ciliocytophtoria without precancerous atypical squamous metaplasia. The development of centrilobular emphysema was also observed. Numerous macrophages within the lumen of bronchi, in the interstitium as well as those within the lung alveoli, contain granules with a positive reaction on copper. A four-month-period is not enough for inducing interstitial fibrosis or granuloma foration within the lung tissue, what some authors have found in experimental animals during their longer exposure in relation to humans, as well as in vineyard sprayers during their longer exposure at work, where cytologically can be found unusual mataplastic bronchi cells. At least a six-month period of exposure influenced by Bordeaux Mixture is thought to be the minimal period needed for development of changes in the lung tissue which can cause "Respiratory Failure" as well as unplastic expansion. PMID- 16268059 TI - Clinical study on safety and efficacy of the administration of amlodipine in a combination with lisinopril in hypertensive patients. AB - AIM: Evaluation of efficacy and safety of long-term (24 weeks) administration of a combined therapy of ACE inhibitors (lisinopril) and calcium-channel antagonists with long-term action (amlodipine) (with or without diuretics). METHOD: A total number of 98 patients of both genders were included in the prospective, open trial. Evaluation of efficacy of the trial was carried out by careful monitoring of anamnesis and physical examination, blood-pressure values, pulse, body weight, ECG test and echocardiographic test. Evaluation of safety of the trial was based on the evaluation of a physician, standard laboratory evaluation, adverse effects. RESULT: Mean value of blood pressure in the beginning of clinical trial was significantly reduced (from 197.65 +/- 18.05/107.1+/-13.1 mm Hg, to 139.85 +/ 10.45/82.6+/-5.47 mmHg) after 24 weeks, without special oscillations in last three months. The pulse remained within physiological limits. In 73.08% patients, echocardiographic test has proven an improvement of ejection fraction. Minor adverse effects have been reported, which were not a reason for interruption of the treatment, with the exception of 3 cases (lower leg oedema, dry cough). CONCLUSION: An extremely good effect in patients with moderate hypertension was reported in the trial, as well as in patients with severe hypertension with good tolerance. PMID- 16268060 TI - [The results of treatment children's Hodgkin lymphoma at pediatric clinic in Sarajevo]. AB - The aim of this study is to estimate the eficacy of citostatic and radiotherapy in treatment of Hodgkin disease (HD) on Hematology-oncology Department of Pediatric Clinic in Sarajevo (Bosnia and Herzegovina). PATIENTS AND METHODS: In retrospective study we followed all patients with HD in the period of 7 years (01.01 1997-31. 12. 2004) Patients were up to 15 years old, and HD was diagnosed and treated at our Clinic. Till 2000 we used UKCCSG HD 9201 protocol treatment with combination of citotstatic and radiotherapy, and after 2000 protocol UKCCSG HD 2000 (ChlVPP / ABVD) was used for treatment HD. RESULTS: We treated 24 children with HD (10 boys-41,66%, 14 girls-58,33). In 7 children (29,1%) we found II A stage of HD, in 4 children (16,6%) II B stage of HD, in 5 children (20,8%) stage III A, in 3 children (12,5%) stage III B, in 2 (8,3%) stage IV A and in 3 children (12,5%) stage IV B. In 5 patients (20,8 %) appeared relapse due to which we used more aggresive citostatic and radiotherapy, and in 3 patients (12,5%) in combination with bone marrow transplantation. One patient (4,1%) died. There are 23 children (95,8%) alive in 1st or 2nd remission phase. Secondary malignancies did not occur in any of patients. CONCLUSION: Although there were significant number of patients diagnosed in high stage of HD (III and IV) the results of therapy HD on Pediatric Clinic in Sarajevo are good. PMID- 16268061 TI - [Pregnancy after laparoscopic treatment of ovarian endometriotic cysts]. AB - OBJECTIVE: The aim of study was to present the optimal laparoscopic treatment of the ovarian endometriotic cysts in patients who had been treated of sterility and to present the success of the sterility treatment. METHODS: Ovarian endometriotic cysts do not have proper reaction to hormonal supressive therapy and surgical treatment is required. Laparoscopic cystectomy of ovarian endometriotic cysts using electrocision was performed in 45 patients who previously underwent laboratory tests specific for that type of patients, as well as sperm analysis of their husbands. At the same time, potency test of the uterine tubes was performed. RESULTS: 34 patients had normal uterine tubes potency and normal semen analysis of their husbands. Out of 34 patients, 12 (35%) spontaneously got pregnant during 12 months following laparoscopic treatment. At the same period of 12 months, in 6 (13,3 %) patients recurrent ovarian cysts were discovered. CONCLUSION: Laparoscopic cystectomy of ovarian endometriotic cysts is efficient method of preserving fertility in certain number of patients. Although the laparoscopic treatment may reduce ovarian reserve, it presents a method of choice in therapy of ovarian endometriotic cysts. PMID- 16268062 TI - [Perinatal mortality at University Hospital Mostar for five years]. AB - AIM: The aim of this study was to present the trend of the rates of perinatal mortality at the University hospital Mostar in the period from 1999 to 2003 and to determine risk factors of perinatal mortality. PATIENTS AND METHODS: Observed group was formed by 110 pregnant women whose infants died in perinatal period at University hospital Mostar in the period from 1999 to 2003 and control group by 107 pregnant women, the same parity and age, whose infants didn't die in perinatal period. The groups were compared acccording to available parameters which determine perinatal outcome. We determined risk factors of perinatal mortality by using case-control study. RESULTS: Mean of perinatal mortality at University hospital Mostar in the observed five year period was 14,71%, range 8,88-19,1%o. A significantly higher number of premature labor (ten times higher), twin pregnancy (p=0.005; chi2 = 7.76), placental abruption (p<0.001; chi2 = 13.84), preeclampsia (p=0.027; chi2 = 4.891), placental insufficiency (p= 0.002; chi2 =9.395), metrorrhagia (p=0.007; chi2 = 7.237), fetal growth restriction (p=0.007; chi2 = 7.237), fetal asphyxia (p=0.017; chi2 = 5.683), true knots in the umbilical cord (p=0.024; chi2 = 5.12) and congenital malformations of fetus (p=0.024; chi2 = 5.12) were in the observed group than in control. 68,04% of all infants which died in perinatal period were premature babies. CONCLUSION: Rate of perinatal mortality at the University hospital Mostar is unacceptable high and doesn't show clear tendency of decreasing in observed period. In order to decrease perinatal mortality rate it is necessary to improve the organization and quality of perinatal care and start with regionalisation and establishing tertiary centers on state level. PMID- 16268063 TI - [Radical cystectomy early postoperative complications and mortality rate]. AB - The aim of this study is to determine mortality rate, intraoperative, early postoperative complications and length of postoperative hospitalization in patients treated with cystectomy carried out for radical or palliative purposes. TESTS AND METHODS: This retrospective study included 57 patients that were treated in the Urological Clinic Clinical Centre University of Sarajevo in period from January 2000 until July 2004, for bladder cancer. RESULTS: Early mortality rate was 3.5%, early postoperative complication rate was 33.2% and it normally included prolonged ileus, wound dehiscence and urinary infection. CONCLUSION: Early mortality and the intraoperative one did not rise when compared to the results indicated in professional literature; early postoperative complications and length of postoperative hospitalization increased. Postoperative mortality and early postoperative complication rates are not statistically in patients over the age of 70 but they depended on concomitant diseases (comorbidity) and general health status of patients. PMID- 16268064 TI - [Complications of operative treatment fractures by osteosynthesis]. AB - In this paper are showen complications as possibility in a work these who usses osteosynthesis as an operative technic. One of the most frequent complication is infection. Infection can imperil as the operative work as longtime healing. The other complications are: disturbed biomechanical relations and fixation on the fracture place, bad preoperative diagnostics, pseudoarthrosis, osteoporosis etc. PMID- 16268066 TI - [Anxiety in acute phase of ischemic stroke and myocardial infarction]. AB - INTRODUCTION: Period after ischemic stroke and myocardial infarction is followed by different psychological reactions. Anxiety represents one possible psychological problem. AIM: To investigate presence of anxiety in patients within period from 48 hours till 15 days after ischemic stroke and myocardial infarction. SUBJECTS AND METHODS: Fourty patients formed analyzed group. Their average age was 65.3 +/- 10.3 years (33-83). Twenty of them were females. Thirty patients formed controled group. Their average age was 57.46 +/- 10.6 years (42 78). Eleven of them were females. Twenty questions self-assessment Zung scales were used for anxiety levels determination, Responses were scored by points, which were summarized as final result. Final score of 50 and more points suggested that anxiety was present. RESULTS: Average self-assessment anxiety score within 48 hours since ischemic stroke was 44.4 +/- 8.8 points, and 15-th day 42.2 +/- 7.7 (p=0.237). In twelve patients (30%) anxiety was found during the first surveying, and in 10 patients (25 %) during the second the surveying. Average self-assessment anxiety score within 48 hours since myocardial infarction was 43.1 +/- 8.3 points, and 15-th day 43.2 +/- 6.7 (p=0.959). In seven patients (23.3%) anxiety was found during first surveying, and in 5 patients (16.7%) during second surveying (p=0.52). CONCLUSION: Anxiety represents important psychological problem for patients after ischemic stroke and myocardial infarction. Aldo number of anxiety patients is larger within 48 hours after ischemic stroke and myocardial infarction, average value in anxiety scale stays unreduced even 15 days after the beginning of the disease. PMID- 16268065 TI - [Prevention of hepatitis C at Center for Haemodialisys of Clinical Centre of University of Sarajevo]. AB - GOAL: The patients with the chronical programm for haemodialisys have the higher risk from getting ill virus hepatitis C in the realtion to the rest population. AIM OF THE WORK: Was the evaluation of the prevalence and incidence of hepatitis C at the Center for Haemodialisys CCU Sarajevo and the effect of the prevalentive measures on the incidence of the serum conversion of hepatitis C, in the period from 2002 till 2004 year. MATERIAL AND METHOD: By the examening is comprehended 155 patients aged 54,58 +/- 14,797 years, with the aproximative length of the haemodialisys 58,9 +/- 53,9 months. Patients at the chronic programm of the bicarbonite haemodialisys taree times per week, and antibodies on the hepatitis C were determined III generation. Also was determined PCR. RESULTS: During the periiod of examination the dialized population was increased also 2002 year the prevalence of hepatitis C was 23,87% (37/155), in 2003 year 29,29% (46/157) and 2004 year the pregalence amounted 26,28% (46/175). Incidence of hepatitis C was significantly decreased in the course of the period of followup and in 2002 year was 16,21%, in 2003 year 13,04%, that in 2004 year would amount 4,34%, that is only in two patients occurred the serum conversion on hepatitis C. CONCLUSION: By applying of the corresponding protocoles and their strict realization (desinfection of the hands, wearing of gloves, apparatus desinfection) and separation of the dialyzed monitors for anti HCV positive and anti HCV negative patients dicreased significantly the hepatitis C incidence in our dialyzed population. The strict application of the preventive measures can completely prevent the speading occurrence of hepatitis C on haemodialisys. PMID- 16268067 TI - [One or more drugs in epilepsy treatment]. AB - Therapeutic options for epilepsy are increasing, and all antiepileptic drugs can produce adverse side effects. Physicians shuld first use monotherapy when treating an epileptic patient. If one drug proves insufficient to control all seizures without intolerable side effects, then careful combination of the fewest drugs possible must be used. Combine drugs with different mechanisms of action. Closely monitor the patient for adverse side effects, which increase when multiple drugs are taken, and measure antiepileptic blood levels more frequently than with patients who are on monotherapy. PMID- 16268068 TI - [Complex functional test in juvenile rheumatoid arthritis]. AB - The Juvenile rheumatoid arthritis is a special form of rheumatoid arthritis which occurs in childhood and adolescence (before the age of sixteen). The disease primarily affects the joints, but can also cause heavy damages to organs and systems such as the heart, blood vessels, skin, epidermis, eyes, peripheral nerves etc. The disease occurs more often with girls. In the therapeutic sense, viewed from the physical medicine and rehabilitation aspect, lying in bed is needed during the acute phase of the disease, and when the inflammation ceases, normal body activities should be encouraged. Rehabilitation with standard methods of physical therapy includes enhancement of daily activities and quality of life, adaptation of patient to the new conditions and prevention of complications. At the Physical medicine and rehabilitation department of the Pediatric clinic CCU Sarajevo, during year 2001 and 2002, fourteen children were observed. All children were treated with kinezitherapy, after which everyday life activities were observed as well as the mobility. In these everyday life activities we were observing whether child can use spoon, fork and knife, whether or not a child can dress his/her upper or lower part of the body with or without buttoning, whether or not it can bathe without someone's help. While observing mobility we evaluated general mobility with changing the body's position from lying to sitting and also from sitting to standing position, climbing onto and getting of a bed, ascending and descending stairs, ability to crouch down or not. These methods are used to prevent inability or decrease it as much as possible. It is accomplished by preventing the forming of muscular atrophies, preventing the development of contractions of joints, as well as preventing the development of osteoporosis and bone destruction due to the inactivity. The imperative of a good rehabilitation is "active movement". PMID- 16268069 TI - [Epidemiological characteristics of low back pain in selected sample of rehabilitation centre "Reumal" in Fojnica]. AB - Low back pain, with or without propagation to lower extremities, still remains challenge not only for therapy but as diagnostic entity as well. This is the reason why literature is regularly refreshed with new epidemiological studies of low back pain. Six years retrospective study included 55.725 patients rehabilitated in Rehabilitation centre "Reumal" in Fojnica, from January 1997 to December 2002. From this sample it has been isolated 12.296 patients with diagnosis referred to low back pain, in order to get certain epidemiological parameters of low back pain, as: intrahospital prevalence, gender and age overview, overview of diagnostic entities referred to low back pain and their relationship to gender. Total prevalence of low back pain was 22,6 %, ranging from 17,8 % to 28,6 % by years. This epidemiological parameter of low back pain, as well as others, correlate with data of other studies. PMID- 16268070 TI - [Viruses in water]. AB - The article elaborates the significance of water, which is necessary for the maintenance of an organism and specifies its daily needs in human beings. Reference data on the early isolations of viruses from sewage water and modern understanding on the occurring virus species are listed. The article considers virus survival in sewage and flowing water and other fresh and marine water systems as well as marine silt or fruit. Finally, we give here an overview of basic features of individual virus kinds that are potential water contaminants. PMID- 16268071 TI - [The level lipids in serum for specific professions]. AB - The high level lipids combine with other factors of risk for atherosclerotic heart disease and stroke (CVA) are one of the most important health problems all over the world. A risk for development of such changes is higher for specific profession as policemen. In this prospective study 300 policemen were medically examined; 150 tested patients and 150 policemen in control group. The same methods were applied to both groups: detailed anamnesis, physical check up, complete laboratory testing, electrophoresis of lipids at hypercholesterolinemia, Ultra-sound examination of abdomen and The Color Doppler analysis of neck arteries. Results obtained by statistical data processing have shown that the level of cholesterol and triglycerides in serums of the tested patients is significantly increased in comparison with the control group. Ultra-sound examination of abdomen detected fatty infiltration of liver for 16% patients and for 2% control group. The Color Doppler analysis of neck arteries for 14% patients revealed from hardly observable changes to dramatic narrowing of lumen up to 50%. For control group this percent was 0,66. Having in mind that this group consists of young active people, the problem of hyperlipidemy becomes even more serious. PMID- 16268073 TI - [Echocardiographic assessment of diagnosis and prognosis of biopsy-proven amyloid cardiomyopathy]. AB - Amyloid cardiomyopathy is myocardial infiltrative disorder which mostly has been seen as the consequence of systemic amiloidosis. The diffuse global myocardial infiltration of nonfunctional amyloid displaces the contractile myocites giving rice to relaxation abnormality and diastolic dysfunction of restrictive or congestive type of both ventricles, but more frequently with right-sided congestion, while systolic left ventricular function deteriorates late in disease process. We report a patient with amyloid cardiomyopathy and nephrotic syndrome underlying primary amiloidosis. Our aim is to point out at echocardiographic assessment of diagnosis and prognosis of amyloid cardiomyopathy, which is proven by postmortal endomyocardial biopsy. The hallmark of echocardiographic diagnostics are the findings of the thickened ventricular and septal walls, small ventricular cavities, dilated atria with thickened interatrial septum and atrioventricular valves, and granular-sparkling and hyperrefractile myocardium. Doppler assessment diagnostically gives us the insight in restrictive physiology of both ventricles, and the inverse relation of the left ventricular thickness and voltage on the ECG is high specific. Echocardiographic evaluation of mean left ventricular thickness in amyloid cardiomyopathy is very important prognostic parameter. so that if it is > or =15 mm, median survival is 0.4 years, whereas in our patient with median thickness of 2.76 cm the survival was only three months. The advanced diastolic dysfunction of the left ventricle with an increased transmitral E/A ratio and deceleration time of < or =150 ms is strong predictor of cardiac death. In this case of restrictive transmitral flow E/A was 1.7 and DT 100 ms and they were ominous prognostic signs of survival. PMID- 16268072 TI - [The role of the team of family physician in prevention of changing risk factors important in development of arterial hypertension]. AB - Arterial hypertension (AH) is one of the commonest noninfective chronic disease according to its important and the role in the morbidity and mortality, which is the reason for patients coming to the family phisician. Detection and treatment of high blood pressure are the major responsibility of physician in the primary care. If the family physician team (physician and nurse) make a good assessment of the risk factors which is important in development of arterial hypertension, the appearance of disease and its complications can be prevented or delayed. The most important for prevention of arterial hypertension is adoption a healthy lifestyle and it is nonseparate part of arterial hypertension treatment. PMID- 16268074 TI - [Uncommon foreign body in the respiratory airways]. AB - Foreign body most frequently reaches respiratory airways by act of aspiration. It could be of various origin, most dangerous is vegetables one which inlarges progressively and could obturate airways completelly. Consequences of that is problem with breathing, atelectasis, and futher on if that obstactle is of long duration bronchiectasis and abscess of pulmonum can be developed. The aim od this paper is to present uncommen foreign body located in the right bronchus- plastic tube. Methods of choice: bronchoscopy and extraction. PMID- 16268075 TI - [Neuropsychiatric lupus and lupus nephritis as a clinical manifestations of systemic lupus erythematosus]. AB - Neuropsychiatric (NP) lupus and lupus nephritis are one of the most profound manifestations of the Systemic lupus erythematosus (SLE), with wide variety of clinical manifestations. Especially NP lupus is the most poorly understood subset of the disease, and the most difficult therapeutic problem. We present case report of female SLE patient with the associated difficult and different clinical manifestations of central and peripheral nervous system disease end renal involvement. Agressive treatment option with intermittent pulsed intravenous cyclophosphamide and corticosteroids after the second month of treatment brought to complete remission of nephrotic syndrome. Improving of life-threating clinical manifestations of NP lupus was obtained after six months treatment by this immunosupressive therapy and included intravenous immunoglobulin 400 mg/kg body weight during five days monthly. PMID- 16268076 TI - [Similarity and difference between American (JNC 7) and European (2003 ESH/ESC) guidelines for management of hypertension]. AB - The American guidelines for management of hypertension (JNC 7) and the European guidelines (2003 ESH/ESC) communicate some new moments and attitudes in the treatment of the condition. According to JNC 7, in patients belonging to the prehypertension category (systolic bp 120-139, diastolic bp 80-89), life style modifications are prescribed. The second and third degrees of hypertension have been merged (degree 2). The drugs of choice for majority of the patients, according to INC 7, are thiazide diuretics, alone or in combination with other drugs. The European guidelines have, more or less, preserved the old classification of hypertension. According to the European guidelines, the assessment of global cardiovascular risk is one of the more important factors in making the decision when to begin the therapy of hypertension. In line with the European guidelines, it is not so important which drug will be applied at the beginning of therapy because, in time, most of the patients will be receiving two or three drugs. PMID- 16268077 TI - [Crisis of the professional ethics at educational system of Bosnia and Herzegovina and its relationship]. AB - In the structure of ruining of the social values system, in post war Bosnia and Herzegovina we possess the worrying degradation of the professional morals in the educational performance: irreal examination, intervention, bribe, sexual black mail and similar. That confirmed the results of the extensive examination which recently was realized on this subject. The negative crisis effects of the professional morals in the educational system will, surely, at the social plan, more long-term reflect. Because, it is no about only in the intelectual-expert, than also about the educational component of personality forming. PMID- 16268079 TI - Implementing the practicum requirement. PMID- 16268078 TI - The Harambee Nursing Center. PMID- 16268080 TI - Courts' perceptions of the responsibilities of nursing practice. PMID- 16268081 TI - Spousal codependency in making end of life decisions: a proposal. PMID- 16268082 TI - Art therapy with family caregivers of patients with cancer. AB - Walsh et al.'s (2004) study supported art therapy to reduce stress, decrease anxiety, and increase positive emotions. The results of this study can be used to support the group's utilization project to promote and educate nurses about the importance of creative arts programs with patients and family members and ways to help establish a creative arts program. One suggestion for future research would be to determine whether the severity of the illness of the patient plays a role in the effectiveness of the art therapy intervention. One feasibility issue for implementing art therapy in hospitals would be the cost of the materials, such as paper, watercolors, markers, and fabrics. PMID- 16268084 TI - Disaster preparedness: students learn about the nurses role. PMID- 16268083 TI - Preceptors: critical link to nursing efficiency. PMID- 16268085 TI - Kentucky's war on weight & type 2 diabetes in youth: the battle continues as a legislative victory emerges. PMID- 16268086 TI - Diabetes standing orders. PMID- 16268087 TI - Planning and emotional health of abused adult children caregivers. AB - The purpose of this study was to explore markers associated with the advance planning patterns and emotional health of adult child caregivers pre-selected for their history of childhood abuse within the family. An informational decision grid and two classification procedures were used to organize and link 246 written plans of 50 adults providing nearly 6 hours of care per day to a cognitively impaired parent. Results revealed 3 constellations of plans describing hands-on person-centred, instrumental, and grief-based approaches to caregiver planning. The 31 participants taking an instrumental approach to planning were significantly more likely than the 17 taking a grief-driven approach to report experiences of abandonment, betrayal, suicidal ideation, and use of alcohol. The 2 participants taking a person-centred approach reported significantly less emotional arousal and risk for loss of health and did not recount experiences of distress or emotion-focused coping. Nurses can use the findings to highlight and strengthen the decision-making and parental-care capacities of caregivers traumatized in childhood. PMID- 16268088 TI - Enhancing health-care research: an interdisciplinary collaborative approach. AB - Many research programs tackle complex problems that cannot be comprehensively investigated by a sole researcher or a research team from a single profession. Interdisciplinary teams can develop a collective mass of common knowledge, broaden the scope of research, and produce more clinically relevant outcomes that are sensitive to the realities of practice. The authors describe the experience of a research team from the perspective of its members. The purposes of the paper are to highlight the benefits of an interdisciplinary collaborative approach to research and to describe the characteristics of a successful team. Some of the benefits discussed include increased research productivity and quality, professional development and mentorship, support and encouragement, expanded resource networks, and bridging of the gap between academia and practice. The authors also discuss the characteristics of a successful research team, associated challenges, and recommendations for enhancing research endeavours through collaboration. PMID- 16268089 TI - Through seniors' eyes: an exploratory qualitative study to identify environmental barriers to and facilitators of walking. AB - This qualitative exploratory study examined environmental factors influencing the walking choices of elderly people using the photovoice approach. A total of 13 seniors in Ottawa, Canada, took photographs of barriers to and facilitators of walking in their neighbourhoods. These photos were displayed during 3 focus-group sessions and served as touchstones for discussion. A total of 22 seniors, including 8 of the 13 photographers, participated in the focus-group sessions. The findings show that environmental hazards related to traffic and falls risks can be significant barriers to walking for seniors, and that connectivity can truly exist for the elderly only if convenience, hazard-free routes are available. They also indicate that simple amenities such as benches and washrooms might facilitate walking for seniors. A neighbourhood that is activity-friendly for seniors will also be a good place for everyone else to live, work, and play. The use of photovoice as a method was well received by the participants and provided rich information that may not have been captured through other means. PMID- 16268090 TI - Unit culture and research-based nursing practice in acute care. AB - The purpose of this multiple-case study of research utilization (RU) was to examine whether and how nursing practices in acute-care units are built on research and to identify potential explanations for the observed patterns. Open ended data were collected from staff nurses and nursing leaders on 8 acute-care units through interviews and observation. RU varied within and across units, but unit culture emerged as the principal factor linked to patterns of RU. Unit culture themes that formed the links were harmony of research perspective, motivation to learn, goal orientation, creativity, critical inquiry, mutual respect, and maximization of resources. The findings provide a rich description that could serve as a basis for self-assessment of unit culture in inpatient and outpatient acute-care units. PMID- 16268091 TI - Categorical service allocation and barriers to care for children with chronic conditions. AB - Parents raising children with chronic conditions face the challenge of locating and coordinating appropriate community-based resources and services for their child. The purpose of this secondary analysis was to determine parents' view of the mechanisms used to allocate health, education, and social services to children with chronic illness and disability and their caregiving families. A thematic analysis was conducted on data from interviews with 30 mothers and 13 fathers. These parents described 11 mechanisms that were used to determine eligibility and/or to ration services: diagnosis, age, technology dependence, severity, functional ability, guardianship status, geographic location, financial resources, judged parental coping, failure to inform parents about available services, and cyclical funding. These mechanisms were complex, inconsistent, and subject to change. Mechanisms that are integrated, proactive, flexible, and fair are needed to reduce parents' workload and to ensure more equitable allocation of services. PMID- 16268092 TI - Determinants of health-service use by low-income people. AB - Poverty influences health status, life expectancy, health behaviours, and use of health services. This study examined factors influencing the use of health related services by people living in poverty. In the first phase, 199 impoverished users of health-related services in 2 large Canadian cities were interviewed by their peers. In the second phase, group interviews with people living in poverty (n = 52) were conducted. Data were analyzed using thematic content analysis. Diverse health-related services were used to meet basic and health needs, to maintain human contact, and to cope with life's challenges. Use of services depended on proximity, affordability, convenience, information, and providers' attitudes and behaviours. Use was impeded by inequities based on income status. To promote the health of people living in poverty, nurses and other health professionals can enhance the accessibility and quality of services, improve their interactions with people living in poverty, provide information about available programs, offer coordinated community-based services, collaborate with other sectors, and advocate for more equitable services and policies. PMID- 16268093 TI - Social and geographical boundaries around senior nurse and physician leaders: an application of social network analysis. AB - The purpose of this study was to describe the social and geographical boundaries around the networks of senior nurse executives and physician leaders and managers in acute-care hospitals in the United Kingdom. A telephone survey was conducted using standard social network methods. A random sample was drawn from a national list and repeatedly sampled until 100 respondents were interviewed. The response rate was 49.5%. Both groups tended to discuss "important professional matters" with others who were similar to themselves in terms of profession, gender, age, and seniority, with physicians being more extreme in this regard. The implication is that gaps in the network of informal ties will impede the dissemination of information and the spread of social influence between these 2 important groups. Managers (non-clinically qualified) appear to occupy a powerful "brokerage" role. Informal networks are mainly composed of local ties. The authors argue that dissemination and influence strategies that take features of the social structure into account are more likely to be successful. PMID- 16268095 TI - Skills enhancement: strengthening public health practice in Canada. PMID- 16268094 TI - Factors to weigh when considering electronic data collection. AB - Researchers are increasingly considering the adoption of electronic data collection methods--which entail the use of the Web, e-mail, and desktop and handheld computers--for surveys, diaries, research instruments, and focus groups. Based on prior research findings on electronic data collection, the authors delineate the key factors, which incorporate population, data, and resource characteristics, that guide researchers in selecting the data-collection method most appropriate for their research question. Population factors to consider in data-collection decisions are age, gender, socio-economic status, and access to technology. Key data factors influencing the selection of collection method include sensitivity of the topic, time sensitivity, longitudinal data, and contextual data. Data-collection decisions are also based on financial, time, and technological resources. Technology and demographics will continue to change and affect data-collection methods and possibilities. By examining these key factors, however, researchers will be able to reach data-collection decisions that are appropriate for each project. PMID- 16268096 TI - Study of survival, dispersal and home range of autumn-released red-legged partridges (Alectoris rufa). AB - 1. On a private property with a stable population of wild red-legged partridge (Alectoris rufa) and an appropriate habitat for the survival of the species, reinforcement repopulations were carried out in the months of October and November for two consecutive years using 5- to 6-month-old birds reared on a commercial game farm. 2. Of the 36 released birds, none was still alive by the following spring's breeding period. Mean survival time was 9.4 d in the first year and 7.6 d in the second year. 3. Seventy-two per cent of mortality was attributable to predation, 11% to hunting and 17% to doubtful causes of death, accidents and starvation. 4. Post-release mean dispersion was 377.8 m in the first year and 526.3 m in the second. Mean home range was 7.1 ha in the first year and 5.4 ha in the second. 5. The production systems and handling practices of commercial game farms may have modified some anti-predator ethological patterns and strategies, which might make it more difficult for the birds to adapt and integrate into the wild, resulting in reduced survival due to premature mortality. PMID- 16268097 TI - Chicken welfare as indicated by lesions on carcases in supermarkets. AB - 1. The extent of visible lesions in British broiler carcases was investigated in order to discover the extent of any poor welfare. It is well established that hock burn and some other injuries can occur during housing of broiler chickens, whilst other injuries can occur during handling. 2. Three hundred and eighty-four whole conventionally reared British Farm Standard Grade A chickens were scrutinised in supermarkets to assess the frequency of 15 kinds of lesions. Six lesions were analysed histopathologically and this showed that hock burn would have occurred several days pre-mortem, red alular and wrist bruise were very likely to have occurred pre-mortem, whilst red parson's nose and scratches were likely to have occurred pre-mortem. 3. Many broiler carcases in the supermarkets had dermal lesions: 0.82 had hock burns, 0.45 a red parson's nose, 0.32 a red alular, 0.21 wrist bruises and 0.21 had at least one scratch on a leg. The frequency of the larger hock burns increased with body weight. 'Organic' chickens had half as many hock burns as conventionally reared broilers, perhaps because of differences in litter quality or leg strength. 4. The sample studied indicates that painful lesions which would result in poor welfare are frequent in broilers slaughtered in the UK. The Grade A chickens observed exclude birds with obvious visible defects because these birds would have had the blemishes removed and the carcase would have been portioned. Hence the frequencies of lesions in farmed birds would be higher than those reported here. PMID- 16268098 TI - Recovery of duodenal villi and cells in chickens refed protein, carbohydrate and fat. AB - 1. To clarify how histological recovery of villi and cells would be affected after refeeding single nutrients such as protein, carbohydrate and fat, male chickens were divided as follows: (1) intact control fed ad libitum a commercial finisher mash diet (CP, 140 g; ME, 11.71 MJ/kg, ALM), (2) 3 d feed withdrawal (FW), (3) FW followed by one day ad libitum free access to the mash diet (FW ALM), and FW followed by one day force-feeding of (4) a commercial finisher pellet diet (FW-FFM) and an isocaloric diet of (5) a protein (FW-FFP), (6) a carbohydrate (FW-FFC) or (7) a fat (FW-FFF). 2. After refeeding, the formula diet groups increased in villus height and villus area and tended to increase in cell area and cell mitosis. Furthermore, flat cells on the villus tip in the F group developed to dome-shaped cells. This suggests that nutritionally well-balanced diets can induce histological recovery at villus and cellular levels. 3. Not all of the single nutrient groups recovered to the extent of the formula diet groups in all light microscopic variables after refeeding, suggesting that a single nutrient cannot induce histological recovery of the villus. 4. However, the dome shaped cells were more distributed on the villus tip in these single nutrient groups than in the well-balanced formula diet groups, although cell diameter of the former groups was smaller than that of the latter. This suggests that the single nutrients would be effectively absorbed from cells and can induce histological recovery at the cellular level. PMID- 16268099 TI - Performance of indigenous, Khaki Campbell and their reciprocal crossbred layer ducks under different management systems. AB - 1. The performance of indigenous ducks (ID), Khaki Campbell (KC) and their reciprocal crossbred layers was studied from 19 to 58 weeks of age. For each genotype, 4 x 18 ducks (3 males + 15 females) were reared under a semi-intensive system (SIS) and an intensive system (IS) with standard management, and 4 x 50 ducks (8 males + 42 females) were reared in an extensive system (ES) with traditional management. 2. In comparison to KC, ID were superior in terms of age at first egg, age at 50% egg production, egg weight, hatchability, eggshell thickness with higher egg shape index. KC ducks were superior to ID in body weight, egg production and feed/kg eggs. Egg quality was similar among the genotypes. Crosses were superior to their parent breeds in age at first egg, egg production and feed/kg eggs. They were also superior to KC in egg weight and egg shell thickness with a higher egg shape index. 3. The performance of genotypes in the SIS and the IS was similar and superior to the ES except for fertility and yolk colour. 4. Significant heterotic effects were recorded for age at first egg, age at 50% egg production, egg production per duck-day, feed efficiency and egg weight in crosses. Performance was similar in the reciprocal crosses, but superior to their parent breeds. PMID- 16268100 TI - Mapping of quantitative trait loci affecting organ weights and blood variables in a broiler layer cross. AB - 1. A genome scan was performed to locate genomic regions associated with traits that are known to vary in birds (most commonly broilers) suffering from heart, lung or muscular dysfunction and for weight of the dressed carcass and some internal organs. 2. The F2 population studied was derived from a cross between a broiler and a layer line and consisted of over 460 birds that were genotyped for 101 markers. 3. There was strong support for segregation of quantitative trait loci (QTL) for carcass and organ weights and blood variables. We identified 11 genome-wide significant QTL (most of them for dressed carcass weight) and several genome-wide suggestive QTL. 4. The results point to some genome regions that may be associated with health-related traits and merit further study, with the final aim of identifying linked genetic markers that could be used in commercial breeding programmes to decrease the incidence of muscular and metabolic disorders in broiler populations. PMID- 16268101 TI - Effect of ochratoxin A on broiler chicks challenged with Salmonella gallinarum. AB - 1. A study was conducted to evaluate the effects of ochratoxin A (OA) on broiler chicks challenged with Salmonella gallinarum. 2. One hundred and seventy-six 1-d old broiler chicks were divided into two groups of 88 chicks each, with one group fed on a control mash diet and the other given a mash diet containing 2 ppm OA. On d 14, each group was further subdivided into two groups with one group infected with S. gallinarum and the other uninfected. 3. Following S. gallinarum inoculation on d 14, 4 birds from each group were killed at 1, 2, 3, 5, 7, 10, 14 and 21 d post inoculation. 4. S. gallinarum infection caused dullness, depression, weakness, increased thirst, droopy wings, ruffled feathers and greenish-yellow diarrhoea. S. gallinarum infection in the absence of OA caused 11.5% mortality which increased to 28.8% in the presence of OA. 5. Decreased body weight and reduced feed intake were observed in chicks fed on the diet containing OA. S. gallinarum infection also reduced the body weights of chicks, with the effects being more marked in chicks receiving OA. The OA diet led to increased serum levels of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, uric acid and creatinine, and decreased levels of total proteins, albumin, globulins, calcium and phosphorus. S. gallinarum infection did not cause significant alteration in any of the serum biochemical parameters. 6. Mortality and the severity of S. gallinarum infection in broiler chicks were increased by the presence of OA in the diet. PMID- 16268102 TI - Effect of chilling, polyphosphate and bicarbonate on quality characteristics of broiler breast meat. AB - 1. An experiment was conducted to assess the effect of tetrasodium pyrophosphate and sodium bicarbonate on colour and sensory attributes of pre- and post-chilled breast meat. 2. Three groups of 6 halves of breasts (pre-chill) immediately after slaughter were treated with 3% tetrasodium pyrophosphate, 3% sodium bicarbonate in 2% NaCl or 2% NaCl alone (control); the remaining 6 halves (post-chill) were stored overnight at 4 degrees C and then treated similarly. Both the pre- and post-chill samples were held at 4 degrees C for 24 h and pH, water holding capacity, cooking loss, CIE colour values and sensory attributes were recorded. 3. Chilling had few effects on the meat characteristics measured in this study. 4. Treatment with phosphate and bicarbonate increased pH in both the pre- and post-chill groups. Treated breasts exhibited lower L* and higher a* value (more red) than controls. 5. A sensory evaluation study revealed improvements in colour and other sensory attributes of cooked broiler breast meat in all treated samples compared to the control. 6. The findings suggest that tetrasodium pyrophosphate and sodium bicarbonate, when injected post mortem, will have beneficial effects on several physico-chemical (pH, colour, WHC %, cooking loss) and sensory attributes of broiler meat. However, phosphate had a smaller effect than bicarbonate. PMID- 16268103 TI - Chromium picolinate, rather than biotin, alleviates performance and metabolic parameters in heat-stressed quail. AB - 1. The effects of chromium picolinate and biotin supplementation alone and in combination on performance, carcase characteristics, malondialdehyde (MDA), vitamin C, vitamin E, glucose and cholesterol levels were evaluated in Japanese quail exposed to high ambient temperature. 2. Two hundred and forty quails (10d old) were assigned randomly to 4 dietary treatments at room temperature (22 degrees C; thermoneutral, TN) or ambient (34 degrees C for 8 h/d; heat stress, HS). Both TN and HS were fed either on a basal (control) diet or the basal diet supplemented with 400 microg of Cr/kg (Cr group), 0.5 mg of biotin/kg of diet (biotin group) or both (Cr + Biotin group). 3. Supplementing the diet of heat stressed quails with chromium picolinate improved live weight gain, feed intake, feed efficiency and carcase traits. Biotin supplementation during TN and HS conditions did not have any beneficial effects on body weight gain, feed intake, feed efficiency or carcase traits. 4. Either in combination or alone, chromium picolinate increased serum concentrations of vitamins C and E, but decreased MDA, glucose and cholesterol concentrations in birds kept at high ambient temperature. There was no difference in vitamins C and E and MDA concentrations between birds given chromium picolinate and birds receiving chromium picolinate plus biotin, while glucose and cholesterol levels were significantly lower in all groups. The lowest concentrations of cholesterol and glucose were found in the combination group under both TN and HS conditions. An interaction between diet and temperature was detected for glucose and cholesterol concentrations. 5. Excretion rates for zinc, iron and chromium were lower in TN groups than in the corresponding HS groups. Supplementing diet with chromium picolinate and chromium picolinate plus biotin decreased excretion of minerals while biotin alone did not effect excretion of minerals. 6. Chromium supplementation, but not biotin supplementation, attenuated the decline in performance and antioxidant status resulting from heat stress. PMID- 16268104 TI - Effect of microbial phytase on production performance of White Leghorn layers fed on a diet low in non-phytate phosphorus. AB - 1. An experiment with 150 White Leghorn layers was conducted to examine the effect of microbial phytase supplementation of low non-phytate phosphorus (NPP) diets on egg production, eggshell quality, bone mineralisation and retention of nutrients at 32-48 weeks of age. 2. Four isonitrogenous and isocaloric diets were formulated to contain 1.2, 1.8, 2.4 and 3.0 g NPP/kg diet with the two lowest NPP (1.2 and 1.8) supplemented with microbial phytase (Biofeed Phytase, India) at 500 FTU per kg diet. Each diet was offered ad libitum to 5 replicates of 5 layers throughout the experiment. 3. Body weight gain was reduced significantly in the layers fed on the 1.2 g/kg NPP diet as compared to those given diets containing 1.8-3.0 g/kg. Addition of phytase to the 1.2 g/kg diet significantly enhanced the body weight and was comparable with those given diets containing 1.8-3.0 g/kg NPP. 4. No additional advantage resulted from enhancing the NPP levels beyond 1.8 g/kg or adding phytase to a diet containing 1.8 g/kg NPP. 5. Hen d egg production, food intake, food efficiency, shell weight, shell thickness, shell strength and tibia strength followed the same trends as above. However, adding phytase to the 1.8 g/kg NPP diet significantly enhanced tibia ash. Egg weight, specific gravity and Haugh units were influenced by neither NPP concentration nor phytase supplementation. 6. Adding phytase to the 1.2 g/kg NPP diet significantly enhanced nitrogen and phosphorus retention. 7. It was concluded that addition of 500 FTU of microbial phytase/kg diet can allow the reduction of NPP content to 1.2g/kg in the layer diet, eliminate inorganic phosphorus supplementation and results in significant reduction of nitrogen and phosphorus excretion without affecting the production performance of layers. PMID- 16268105 TI - Evaluation of phytase concentration needed for growing-finishing commercial turkey toms. AB - 1. Growth performance, serum bone markers, and bone strength and mineralisation were determined in tom turkeys grown from 9 to 17 weeks of age. 2. Dietary non phytate phosphorus was formulated to be reduced by 1.0 g/kg in the low phosphorus diet compared to a control diet and phytase was added to provide 0, 150, 300, 450 or 600 units/kg activity to the low phosphorus diet. 3. From 9 to 12 weeks of age, body weight and gain:food were reduced by the low phosphorus diet without added phytase, compared to the adequate phosphorus diet. Increasing the concentration of phytase linearly increased these growth parameters. There were no significant growth responses at 17 weeks of age. 4. Serum osteocalcin was reduced by increasing dietary phosphorus at 12 weeks of age when growth was affected, but not at later ages. Serum pyridinoline was reduced by higher dietary phosphorus and decreased linearly with increasing phytase activity at 17 weeks of age. 5. Fracture force of the ulna and femur increased linearly with increasing phytase activity but bone strength was not affected when corrected for bone cross sectional area. Bone strength of the ulna and ash concentration of the ulna and tibia were increased by higher dietary phosphorus. Humerus and ulna ash increased linearly with increasing phytase activity. 6. Water-soluble phosphorus content of the litter was increased by higher dietary phosphorus and addition of phytase to the low phosphorus diet. The increase in water-soluble phosphorus content of the litter when phytase was fed may indicate that phosphorus could be fed at a lower concentration than used in this trial, at least in the finisher diet when phytase is added to the food. 7. Bone fracture force, strength and ash were generally optimised when 450 units/kg phytase activity was added to the low phosphorus diet. However, growth performance was best in the grower I (9 to 12 weeks) phase when 600 units/kg phytase was added to the diet. PMID- 16268106 TI - Dietary tryptophan effects on growth and stress responses of male broiler chicks. AB - 1. Three experiments were conducted to determine growth of broiler chicks fed on test diets formulated to be deficient or adequate in tryptophan (Trp) using gelatin by-product as a means of generating a Trp deficiency. Growth response estimates of broiler chicks to graduations of Trp were determined by dose response criteria and regression analyses. Experiments were conducted using broiler chicks from 1 to 20 d of age. 2. Broiler chicks fed Trp-deficient diets had poor body weight gain, feed intake and feed conversion. Recommended total Trp needs were 2.0, 2.1 and 2.2 g/kg for feed intake, body weight gain and feed conversion, respectively. 3. Blood plasma Trp exhibited a sigmoidal trend while blood plasma glucose increased in a linear manner to supplemental Trp. Physiological stress variables measured were unaffected by dietary Trp. PMID- 16268107 TI - Use of active substances of plant origin in chicken diets based on maize and locally grown cereals. AB - 1. A total of 336 Hubbard Hi-Y broiler hybrids were fed from d 1-41 on diets based on maize or wheat and barley which were supplemented (or not) with 100 mg/kg plant extract consisting of capsaicin, cinnamaldehyde and carvacrol. 2. Body weight was not enhanced, but feed conversion improved by 4.2% on the maize diet and 2.0% on the wheat and barley diet. 3. Plant extract addition enhanced the breast muscle proportion of empty body weight by 1.2% in comparison to control birds. 4. The apparent ileal digestibility of nutrients (crude protein, fibre and amino acids) was not significantly better in birds fed on supplemented diets than in the control group. 5. Reduction of E. coli, Clostridium perfringens and fungi and increase of Lactobacillus spp. were observed in the 41-d-old chickens from the supplemented groups. 6. An increase in the lipase activity in pancreas and intestine wall was found in older birds fed on the plant extract supplemented diets. PMID- 16268108 TI - Bacillus cereus var. toyoii and Saccharomyces boulardii increased feed efficiency in broilers infected with Salmonella enteritidis. AB - 1. The effect on feed efficiency of two probiotics, one prepared with Saccharomyces boulardii and the other with Bacillus cereus var. toyoii, was tested in broilers infected with Salmonella enteritidis. 2. One-day-old chicks were divided at random into three groups and fed commercial feed devoid of antibiotics: group 1 was fed with non-supplemented feed, group 2 was supplemented with S. boulardii and group 3 with B. cereus. At 14 d of age the animals were challenged by the oral route with 1 x 10(7) viable S. enteritidis. 3. At d 47, average live weights were: group 1, 1.77 kg, group 2, 1.89 kg and group 3, 2.06 kg, and were significantly different. Feed conversion rates were 2.61 for group 1, 2.35 for group 2 and 2.30 for group 3. 4. We conclude that both probiotics improved feed efficiency in broilers. PMID- 16268109 TI - Performance of broiler chickens given whey in the food and/or drinking water. AB - 1. The effects on food intake and weight gain of offering broiler chickens (2 to 7 weeks of age) dry food, wet food, wet food containing whey, whey as drinking liquid and combinations of two of these were studied in 5 experiments. 2. Wet feed generally improved both weight gain and feed efficiencies significantly. Feeding whey also improved weight gain and feed conversion efficiency, but whey offered as a drinking fluid had an adverse effect on broiler performance. 3. When whey was offered both as drinking liquid and added to the food it had a deleterious effect. 4. When whey was offered from 4 or 6 weeks of age, it had a better effect than when offered from 2 weeks of age. 5. There was better performance when whey in the drinking water was diluted and/or offered on alternate days or half-days. 6. Broilers allowed to choose between wet and dry feed when water was freely available chose mostly dry feed; in the absence of drinking water they chose mostly wet food. Birds offered water and liquid whey avoided whey completely. 7. It is concluded that whey can be used in diets for broiler chickens by incorporating it in the food as long as drinking water is offered ad libitum. Whey may be offered as a drink if the food is mixed with 1.8 times its weight of water but it is better to dilute the whey with an equal volume of water whether it is added to food or given as drink. Good results can also be obtained when undiluted whey is offered alternately with water, either in half-day or full-day periods. PMID- 16268110 TI - Light intensity can influence plasma FSH and age at sexual maturity in domestic pullets. AB - 1. Shaver White and ISA Brown pullets were reared to 140 d in cage groups of 8 on a 10-h photoperiod of incandescent light and maintained at an illuminance of 3 or 25 lux, or transferred from 3 to 25 lux or from 25 to 3 lux at 63 or 112 d of age. 2. Plasma follicle stimulating hormone (FSH) concentration at 63 and 112 d was higher in both breeds for pullets maintained at an illuminance of 25 lux compared with 3 lux. After 2-4 d, and relative to constant-illuminance controls, plasma FSH increased significantly for ISA Brown transferred from 3 to 25 lux at 63 d and for Shaver White transferred at 112 d. Irrespective of genotype, plasma FSH for pullets given a decrease in illuminance at 63 or 112 d showed a tendency for less change than did constant-illuminance controls. 3. There was no significant difference in sexual maturity for ISA Brown maintained on 3 or 25 lux, but Shaver White pullets exposed to constant 3 lux matured later than those maintained on 25 lux. Shaver White matured later following an increase from 3 to 25 lux at 63 and 112 d, and earlier subsequent to a decrease from 25 to 3 lux at 112 d. ISA Brown pullets were not significantly affected by a change in illuminance at 63 or 112 d, though their responses were in the same direction as Shaver White. 4. Changes in plasma FSH in the 2- to 4-d period following a change in illuminance at 63 or 112 d were not significantly correlated with sexual maturity. PMID- 16268111 TI - Characterisation of glucose transporter (GLUT) gene expression in broiler chickens. AB - 1. Glucose transporter (GLUT) proteins, one of which is the major insulin responsive transporter GLUT4, play a crucial role in cellular glucose uptake and glucose homeostasis in mammals. The aim of this study was to identify the extent of mRNA expression of GLUT1, GLUT2, GLUT3 and GLUT8 in chickens intrinsically lacking GLUT4. 2. GLUT1 mRNA was detected in most tissues of 3-week-old broiler chickens, with the highest expression measured in brain and adipose tissue. GLUT2 was expressed only in the liver and kidney. GLUT3 was highly expressed in the brain. GLUT8 was expressed ubiquitously, with expression in kidney and adipose tissue relatively higher than that of other tissues. 3. Expression levels of GLUT isoforms 1, 3 and 8 in skeletal muscle tissue were very low compared to the other tissues tested. 4. [3H]Cytochalasin B binding assays on tissue from 3-week-old chickens showed that the number of cytochalasin B binding sites in skeletal muscle plasma membranes was higher than in liver plasma membranes. These results suggest that GLUT proteins and/or GLUT-like proteins that bind cytochalasin B are expressed in chicken skeletal muscles. 5. It is proposed that GLUT expression and glucose transport in chicken tissues are regulated in a manner different from that in mammals. PMID- 16268112 TI - Effects of chitosan on growth performance and energy and protein utilisation in broiler chickens. AB - 1. Two experiments were conducted to investigate the effects of dietary chitosan on growth performance, energy availability and protein retention in broilers. 2. Experiment 1 was a 42-d growth assay, in which 294 1-d-old male broilers were given one of 7 dietary treatments. A control feed was supplemented with 5 levels of chitosan (0.2, 0.5, 1.0, 3.0 and 5.0 g/kg) or 50 mg/kg chlortetracycline (CTC). 3. Increasing chitosan inclusion gave a nonlinear increase (P< 0.001) in feed conversion efficiency (FCE). Optimal growth and feed conversion were obtained with 0.5-1.0 g/kg chitosan. 4. In experiment 2, 42 1-d-old male broilers (6/treatment) were individually housed but fed on the same diets as in experiment 1. Excreta were collected from d 19-21 and d 40-42. 5. The addition of 0.5-1.0 g/kg chitosan increased nitrogen retention compared with the control group (P< 0.01), while apparent metabolisable energy in the diets was not altered. PMID- 16268113 TI - MYCOTOX and aflatoxicosis in quails. AB - 1. This study was to evaluate the toxic effects of aflatoxin (AF) on growth performance of quail, and to determine the preventive efficacy of MYCOTOX (oxicinol, tymol, micronised yeast). 2. One hundred and eighty 1-d-old quail (Coturnix coturnix japonica) of both sexes were weighed and randomly divided into 4 experimental groups each with 5 replicates of 9 birds. 3. There were 4 dietary treatments: (1) control with 0 mg AF/kg diet and 0% MYCOTOX; (2) 0 mg AF/kg diet and 0.5% MYCOTOX; (3) 2.5 mg AF/kg diet and 0% MYCOTOX; (4) 2.5 mg AF/kg diet plus 0.5% MYCOTOX. The chicks were maintained on these treatments to 3 weeks of age. Quail consumed the diets and water ad libitum. 4. Body weight (BW) gains in groups receiving AF alone were the lowest at all periods. Feed intake was lowest in the group consuming the AF diet. The addition of MYCOTOX to the AF diet did not prevent or reduce the toxic effects of AF on feed intake at any time period. Feeding diets containing MYCOTOX alone did not change feed intake significantly. With the exception of the 1 to 7 d period, feed conversion of chicks fed the AF diet was similar to those of the other experimental groups. 5. Bursa of Fabricius weight decreased, whereas the relative weights of liver, kidney and spleen increased in quail consuming diets containing AF and AF plus MYCOTOX. Liver colour was normal in the control and MYCOTOX alone group, but was lighter in groups fed AF. 6. The results indicated that MYCOTOX was not effective in preventing the deleterious effects of AF. PMID- 16268114 TI - Exhaled nitric oxide: sources of error in offline measurement. AB - Delayed offline measurement of exhaled nitric oxide (eNO), although useful in environmental and clinical research, is limited by the instability of stored breath samples. The authors characterized sources of instability with the goal of minimizing them. Breath and other air samples were stored under various conditions, and NO levels were measured repeatedly over 1-7 d. Concentration change rates varied positively with temperature and negatively with initial NO level, thus "stable" levels reflected a balance of NO-adding and NO-removing processes. Storage under refrigeration for a standardized period of time can optimize offline eNO measurement, although samples at room temperature are effectively stable for several hours. PMID- 16268115 TI - Acute effects of noise on blood pressure and heart rate. AB - The authors assessed the acute effects of exposure to noise on systolic and diastolic blood pressure, and heart rate, among 46 workers in a midwestern auto assembly plant. Workers wore ambulatory blood pressure monitors and personal noise dosimeters during one work shift. After adjustment for covariates of cardiovascular function, systolic and diastolic blood pressure, along with heart rate, were shown to be significantly positively associated with noise exposure. Although the long-term effect of these associations is not known, results from other studies suggest that they may be harmful. Replication of this study with a larger number of subjects, monitored for several days and in a variety of work settings, is recommended to verify these findings. The results of this and other studies imply that reducing acute noise exposure reduces cardiovascular stress. PMID- 16268117 TI - Somatic and psychological characteristics of noise-sensitive adults in Finland. AB - The authors examined the relationship of noise sensitivity with health status and psychological factors in individuals <70 yr of age in Finland. Subjects (n = 1,355) were selected from a 1988 case-control study, based on the Finnish Twin Cohort, that assessed noise sensitivity, lifetime noise exposure, and hypertension. Other health status and psychological factors were obtained from a questionnaire that had been administered to the same individuals in 1981. Statistical analysis showed that noise sensitivity was associated significantly with hypertension, emphysema, use of psychotropic drugs (i.e., sleeping pills, tranquilizers, and pain relievers), stress, smoking, and hostility, even after adjustment for lifetime noise exposure. These results indicate that noise sensitivity has both psychological and somatogenic components. PMID- 16268116 TI - Environmental exposure and fingernail analysis of arsenic and mercury in children and adults in a Nicaraguan gold mining community. AB - Gold mining can release contaminants, including mercury, into the environment, and may increase exposure to naturally occurring elements such as arsenic. The authors investigated environmental and human tissue concentrations of arsenic and mercury in the gold mining town of Siuna, Nicaragua. The study involved 49 randomly selected households in Siuna, from whom a questionnaire along with environmental and fingernail samples were collected. Environmental samples indicated that mercury concentrations in drinking water, although generally low, were higher near the mine site. Arsenic concentrations were elevated in water and soil samples, but their distribution was unrelated to the mining site. Mercury concentrations in fingernail samples were correlated with residential proximity to the mine, drinking water concentrations, occupation, and, among children, with soil concentrations. Fingernail arsenic concentrations correlated with drinking water concentrations among adults who consumed higher levels, and with soil concentrations among children. Fingernail analysis helped to identify differential exposure pathways in children and adults. Mercury and arsenic uptake via soil exposure in children warrants further consideration. PMID- 16268119 TI - An evaluation of residual organochlorine pesticides in popular Indian herbal teas. AB - Herbal preparations are gaining popularity worldwide because of their history of use and the belief that they are free of harmful side effects. Among the most popular products are herbal teas, which are marketed extensively with emphasis on their medicinal properties. At the same time, the World Health Organization has been emphasizing the need for quality assurance of herbal products, including testing for inadvertent contamination. The authors conducted a quality-assurance evaluation of residual organochlorine pesticides in some popular brands of Indian herbal teas. Organochlorine pesticide residue build-up from agricultural or storage practices was estimated with gas-liquid chromatography. The results revealed scant presence of dichlorodiphenyltrichloroethane (DDT) or its metabolites; endosulfan--a highly toxic pesticide--was absent in all 8 brands of herbal teas studied. Hexachlorocyclohexane isomers were detected in 2 samples, but levels were below the permissible limit for pesticide residue in foods, as promulgated by the Codex Alimentarius Commission. The authors believe that all herbal preparations should be checked for toxic chemical residues to allay consumer fears of exposure to known neurotoxicant pesticides and to aid in promoting global acceptance of these products. PMID- 16268118 TI - Effect of pesticide exposure on acetylcholinesterase activity in subsistence farmers from Campeche, Mexico. AB - The authors surveyed agricultural production methods and pesticide use among subsistence farmers (campesinos) in 4 rural communities of Campeche, Mexico. Self reports of symptoms of poisoning resulting from occupational pesticide exposure were elicited by questionnaire (N = 121), and acetylcholinesterase (AChE) activity during insecticide use was evaluated from blood samples (N = 127). In individuals from 2 of the 4 communities, AChE activity was significantly lower (p < 0.05) than the mean of activity determined for individuals in a reference group. Results of this study show that erythrocyte AChE inhibition provides a good biomarker of exposure to organophosphate pesticides in field studies with human populations. Carbamates, particularly carbofuran, seem to be more associated with exuberant and diversified symptomatology of pesticide exposure than organophosphates. Studies in field communities where both carbamates and organophosphates are suspected to exist should include blood AChE determinations, symptomatology surveys, and socioeconomic questionnaires. The authors recommend that the Mexican National Health Ministry authorities specify additional provisions regarding the use of protective equipment and the adoption of other safety practices during field work, increase information campaigns about the risks of pesticide use and the value of safety practices, and increase programs of medical monitoring and assistance for rural communities dealing with pesticides. PMID- 16268120 TI - Molecular properties and antibacterial activity of the methyl and ethyl ester derivatives of ampicillin. AB - Ampicillin is a beta-lactam antibiotic that is effective against gram-negative bacteria. Ampicillin has a single carboxyl group (-C(O)OH) within its structure which is suitable for forming ester compounds. Diazomethane and diazoethane were utilized to react with ampicillin to form the methyl and ethyl esters, respectively. The ester derivatives of ampicillin were solubilized together (mole ratio 1:1) in LB media and penicillin resistant Escherichia coli added to measure antibacterial activity. Growth inhibition of bacteria was monitored by optical density after a known time period and with known specific concentrations of the ampicillin esters present. Significant growth inhibition of penicillin resistant bacteria occurred at concentrations of the combined methyl and ethyl ampicillin esters from less than 50 microgram/mL to more than 150 microgram/mL. Molecular properties of the ester compounds were determined. The two ester derivatives showed values of Log BB, Log P, polar surface area, intestinal absorption, and solubility suitable for clinical application. The two ester compounds showed zero violations of the Rule of 5 indicating good bioavailability. The two ester derivatives showed greater intestinal absorbance and greater penetration of the blood brain barrier than the parent ampicillin. Favorable druglikeness was determined for both ester derivatives. PMID- 16268121 TI - Role of alpha-tocopherol in the regulation of mitochondrial permeability transition. AB - We previously showed that Ca2+-induced cyclosporin A-sensitive membrane permeability transition (MPT) of mitochondria occurred with concomitant generation of reactive oxygen species (ROS) and release of cytochrome c (Free Rad. Res.38, 29-35, 2004). To elucidate the role of alpha-tocopherol in MPT, we investigated the effect of alpha-tocopherol on mitochondrial ROS generation, swelling and cytochrome c release induced by Ca2+ or hydroxyl radicals. Biochemical analysis revealed that alpha-tocopherol suppressed Ca2+-induced ROS generation and oxidation of critical thiol groups of mitochondrial adenine nucleotide translocase (ANT) but not swelling and cytochrome c release. Hydroxyl radicals also induced cyclosporin A-sensitive MPT of mitochondria. alpha Tocopherol suppressed the hydroxyl radical-induced lipid peroxidation, swelling and cytochrome c release from mitochondria. These results indicate that alpha tocopherol inhibits ROS generation, ANT oxidation, lipid peroxidation and the opening of MPT, thereby playing important roles in the prevention of oxidative cell death. PMID- 16268123 TI - Temperature distribution and electrical properties along the Oriental hornet body. AB - The hornet is an endothermic insect. Daily variations in hornet surface temperature were measured. Three peaks were found between 9:30 and 10: 30 a.m., 11 and 12 a.m. and between 2 and 3 p.m. Electrical current and voltage values were highest along the head. Electrical current along the gaster and the head flowed towards the thorax, i.e., from body parts with minimal temperature towards the body part with maximal temperature. Current and voltage values measured across the cuticle of the gaster were about 5nA and 100 mV, respectively, and these were of the same order of magnitude as the current and voltage values along the cuticle. It was found that: 1) temperature regulation most probably originates in the thorax and 2) there is a correlation between the temperature distribution along the hornet body surface and levels of the cuticular electrical signals. PMID- 16268122 TI - Phosphate deposition during and after hypokinesia in phosphate supplemented and unsupplemented rats. AB - The objective of this study was to show that prolonged restriction of motor activity (hypokinesia) could reduce phosphate (P) deposition and contribute to P loss with tissue P depletion. To this end, measurements were made of tissue P content, P absorption, plasma P levels, urinary and fecal P excretion of rats during and after hypokinesia (HK) and daily phosphate supplementation. Studies were conducted on male Wistar rats during a pre-hypokinetic period, a hypokinetic period and a post-hypokinetic period. All rats were equally divided into four groups: unsupplemented vivarium control rats (UVCR), unsupplemented hypokinetic rats (UHKR), supplemented vivarium control rats (SVCR) and supplemented hypokinetic rats (SHKR). Bone and muscle P content, plasma intact parathyroid hormone (iPTH) levels, P absorption, plasma P levels and urinary and fecal P excretion did not change in SVCR and UVCR compared with their pre-HK values. During HK, plasma P levels, urinary and fecal P excretion increased significantly (p<0.05) while muscle and bone P content, P absorption and plasma iPTH levels decreased significantly (p<0.05) in SHKR and UHKR compared with their pre-HK values and the values in their respective vivarium controls (SVCR and UVCR). During the initial 9-days of post-HK, plasma, urinary and fecal P levels decreased significantly (p<0.05), and plasma iPTH levels, muscle and bone P levels remained significantly (p<0.05) depressed in hypokinetic rats compared with their pre-HK values and the values in their respective vivarium control rats. By the 15th day, these values approached the control values. During HK and post-HK, changes in P absorption, plasma iPTH levels, and P levels in muscle, bone, plasma, urine and feces were significantly (p<0.05) greater in SHKR than in UHKR. Decreased tissue P content with increased P loss in animals receiving and not receiving P supplementation demonstrates decreased P deposition during HK. Higher P excretion with lower tissue content in SHKR and UHKR demonstrates that P deposition is decreased more with P supplementation than without. Because SHKR with a lower tissue P content showed higher P excretion than UHKR it was concluded that the risk of decreased P deposition with greater tissue P depletion is inversely related to P intake, that is, the higher the P intake the greater the risk for decreased P deposition and the greater tissue P depletion. It was shown that P (regardless of the intensity of its tissue depletion) is lost during HK unless factors contributing to the decreased P deposition are partially or totally reversed. It was concluded that dissociation between (decreased) tissue P content and (increased) P uptake indicates decreased P (absorption and) deposition as the main mechanisms of tissue P depletion during prolonged HK. PMID- 16268124 TI - Communication by electrical means in social insects. AB - Social insects, belonging to the order Hymenoptera, maintain a fixed, optimal temperature in their nest. Thus, in social wasps and hornets, the optimal nest temperature is 29 degrees C, despite the fact that they are distributed in regions of varying climates both in the northern and southern hemispheres of the globe. Since hornets and bees are relatively small insects, determination of their own body temperature as well as that of their nest and the brood was made via thermometers or by the use of infrared (IR) rays. It has been suggested that thermoregulation in social insect colonies is effected primarily by the adult insects via muscle activation, that is, fluttering of their wings, which can raise both their own and the ambient temperature by many degrees centigrade. However, the larval brood can also contribute to the thermoregulation by acting as heat resources and thereby raising the ambient temperature by 1-2 degrees C. To this end, the adult hornets are endowed with a well-developed musculature and their larvae, too, have muscles that enable them to move about. Not so the hornet pupae which are enclosed in a silk envelope (the cocoon), with a rather thick silk cap spun by the pupating larvae, and have rather undeveloped muscles. In the latter instance, it stands to reason that the pupae benefit from the nest warming achieved primarily by the adult hornets, but how is the information regarding their thermal needs relayed from them to the adults? Previously we showed that the adult hornets are attracted to the pupae by pheromones released by the latter, but such chemical compounds can only convey information of a general nature and we are still left with the question as to how the adult hornet can gauge or ascertain the temperature of a single insulated pupa. The present study provides evidence that the hornet pupa can indeed transmit information regarding its body temperature via electrical means. PMID- 16268125 TI - How much water is made "non-free" by 36% native hemoglobin? AB - At equilibrium, the concentration ratio of poly(ethylene gycol) (PEG-4000) in a dialysis sac containing a 35.1% solution of native bovine hemoglobin over that in the external solution is 0.196 +/- 0.028 (mean +/- SD). This apparent equilibrium distribution constant or rho-value of 0.196, when viewed side-by-side with the near-equal distribution of sucrose and raffinose in similar native-hemoglobin dominated water suggests all (rather than 80%) of the water in this solution has been altered by the native hemoglobin and is no longer free liquid water. Based on Ling's equation for solute exclusion, we found that an excess of water-to water interaction energy of a mere 4.25 cal/mole could account for both the observed exclusion of PEG-4000 and non-exclusion of sucrose and raffinose. Finally, the long-range action of (even this relatively inactive) native hemoglobin on the dynamic water structure was compared with the exclusion of coated latex microspheres from the altered water 100 microm from the surface of polyvinylalcohol gel (Zheng and Pollack) --in the light of Ling's new theory of ad infinitum water polarization-orientation (under idealized conditions) first publicized at the Gordon Research Conference on "Interfacial Water in Cell Biology" on the campus of the Mount Holyoke College in June 2004. PMID- 16268126 TI - The impact of parental attachment and feelings of isolation on adolescent fear of crime at school. AB - While scores of researchers have examined the antecedents of fear of criminal victimization among adults, research examining the correlates of such fear among adolescents, particularly in the school setting, is limited. Using data from 2136 public school students from a rural Southern state, we examine the association between fear of criminal victimization and race, gender, age, attachment to parents, feelings of isolation, and victimization. We determine that adolescents who have been victimized by crime are far more fearful than their counterparts who have not. Additionally, we determine that youth who have lower levels of attachment to parents and higher levels of isolation/alienation are also more fearful of criminal victimization than their counterparts. Interestingly, the impact of isolation on fear of criminal victimization is stronger for whites than nonwhites while the impact of parental attachment is stronger for males than females. Implications for policy and future research are also discussed. PMID- 16268127 TI - Success of Taiwanese mothers in guiding adolescents. AB - Education for parents was recently mandated in Taiwan and presents a challenge to the schools. The purpose of this study was to determine how two generations perceive parenting strengths and learning needs. Taiwanese mothers of 10- to 14 year-olds (n=209) and their adolescent children (n=201) completed the Parent Success Indicator. Generational reports were compared, and effects of independent variables were examined. The amount of time mothers spent talking to and doing things with their adolescents had the greatest influence on how both groups rated mother success. Unfavorable ratings expressed by mothers and adolescents identified topics that would be appropriate for parent education. The findings will be used by educators and researchers to support parent development in Taiwan. PMID- 16268128 TI - The role of ego-identity status in mating preferences. AB - This study was designed to examine the role ego-identity plays in the mating preferences of late adolescents. In addition to examining the variance in mating preferences explained by ego-identity status, it was hoped that the results could assist in testing the competing Sexual Strategies (Buss & Schmitt, 1993) and Social Role (Eagly & Wood, 1999) theories. Ego-identity and the sex of the participant accounted for a significant amount of variance in the number of sexual partners desired and the penchant for short-term mating. The sex of the participant was the lone predictor of the importance placed on the mate characteristics of physical attractiveness and earning capacity with females placing more emphasis on the former and males placing more emphasis on the latter characteristic. PMID- 16268129 TI - An exploration of adolescent emotional intelligence in relation to demographic characteristics. AB - Emotional intelligence (EI) was measured in 200 youth ages 16-19. EI scores were compared to demographic characteristics of the individuals (age, sex, household income, parents' level of education, and location of residence). Findings indicate that EI levels were positively related to females, parents' education, and household income. The study did not show significant relationships between adolescent EI and location of residence or age. EI scores were significantly different between females and males, with females reporting higher EI levels. A one-way ANOVA showed no significant differences between EI scores and age, location of residence, and household income. Significant differences were found based upon EI scores for parents' education; as they increased, so did EI levels. In a linear regression model, with demograpics as the independent variables and EI as the dependent variable, father's education and sex were both predictors. The results will guide future studies to determine the factors behind adolescent El formation and development. PMID- 16268130 TI - The nature of connections: young fathers and their children. AB - The purpose of this study was to examine the involvement of young fathers with their children at entry to a fatherhood program and at subsequent follow-up. Thirty-eight young fathers participated in this analysis. Using open-ended questions at intake and subsequent follow-up, they were asked to describe in their own words their relationships with their children. A thematic analysis was used to explore their self-reported statements. Three themes emerged from the analysis: Positive Emotionality, Accessibility, and Engagement. A comparison between thematic categories at intake and follow-up revealed that the most dramatic shift occurred in regard to Engagement. A third of the young fathers shifted from describing Positive Emotionality at intake to the more active process of Engagement at follow-up. The area that showed little change between intake to follow-up was related to Accessibility. These findings may suggest that for young fathers, becoming more involved with their children is a process which can be facilitated by participating in a fatherhood program. PMID- 16268131 TI - Correlates of adolescent assertiveness with HIV avoidance in a four-nation sample. AB - Adolescents are frequently admonished to be socially assertive in order to confront negative interpersonal peer influences. Since the advent of HIV/AIDS in human social chemistry, the admonition has become more critical than ever. But the warning is often proffered in the misguided presumption that social assertiveness is the all-required psycho-structure against risk. The present cross-national study examines social and sexual assertiveness in four-country adolescent samples of Nigerian, Thai, Chinese, and Japanese college students. The findings reveal that sexual rather than social assertiveness was the sine qua non against risk, cross-culturally. PMID- 16268132 TI - Demography, psychosocial factors, and emotional problems of Korean American adolescents. AB - This study attempted to identify emotional problems and examine the related demographic and psychosocial factors of 340 Korean American adolescents in a major metropolitan area. Results revealed that lower GPA, longer length of residence in the United States, subjects' poor self-esteem, greater severity of conflict with parents, and poor relationships with peers were significant predictors of emotional problems. The findings could be beneficial for social work practitioners and educators in determining the service needs of Korean American adoelscents and designing interventions for their problems. PMID- 16268133 TI - The effect of individual differences on adolescents' impulsive buying behavior. AB - This study posits a relationship between Emotional Intelligence (EI) and Impulsive Buying Tendency (IBT). A survey of 574 adolescents found that high-EI adolescents manifested less impulsive behavior than did low-EI adolescents, and high-IBT adolescents were more likely to engage in more impulsive buying behavior than were low-IBT adolescents. Finally, possible extensions of the research to the area of adolescents' impulsive buying are suggested. PMID- 16268134 TI - The role of personality factors in predicting the reported physical health symptoms of Turkish college students. AB - This study investigates the role of optimism, health control beliefs, perceived health competence, and medical help-seeking variables in predicting the frequency of reported physical symptoms. A total of 345 college students (207 male and 138 female) were presented with the Life Orientation Test, Multidimensional Health Locus of Control, Perceived Health Competence Scale, and Physical Symptom Checklist. Separate stepwise multiple regression analyses were applied to the data obtained from males, females, and the total group of students. Results showed that optimism, medical help seeking, chance health locus of control, and internal health locus of control predicted the reported physical symptom scores of the total sample. Also revealed were the different patterns for males and females. While optimism and internal health locus of control best predicted physical health for males, optimism and chance health locus of control best predicted the physical health symptoms of females. PMID- 16268135 TI - Adolescents at risk for violence: an initial validation of the life challenges questionnaire and risk assessment index. AB - Initial validation was sought for the Life-Challenges Questionnaire-Teen Form, a 120-item youth-risk assessment tool. The questionnaire was administered to 99 students enrolled in an adolescent detention facility and a comparison group of 305 students attending high school. The survey items included correlates of youth violence and categorized risk level in a Risk Assessment Index (RAI) based on 53 critical items most strongly correlated with youth violence. Higher RAI scores were expected for the detention sample, males, minorities, and 15- to 18-year olds. Differences between adolescents in detention and high school in terms of risk for violence were assessed by means of analysis of variance, and multiple regression analysis was used to examine the relative effect of detention status, race/ethnicity, gender, and other factors on risk behavior as measured by the RAI. Findings revealed that the detention group endorsed correlates of youth violence more often than the non-detention group and received significantly higher RAI scores. In addition, being in detention, male, and a racial/ethnic minority were significant predictors of risk behavior. The authors conclude that The Life-Challenges Questionnaire (and Risk Assessment Index) effectively differentiated between the detention and student samples, thus, providing initial support for its validity as a risk-assessment measure. PMID- 16268136 TI - The relation of facial affect recognition and empathy to delinquency in youth offenders. AB - Associations among facial affect recognition, empathy, and self-reported delin quency were studied in a sample of 29 male youth offenders at a probation placement facility. Youth offenders were asked to recognize facial expressions of emotions from adult faces, child faces, and cartoon faces. Youth offenders also responded to a series of statements on emotional empathy, and provided self reported acts of delinquency. Findings revealed a moderate positive relationship between ability to recognize the expression, angry, in adult faces, and self reported acts of delinquent behavior, which included physical violence, theft, and vandalism. Findings revealed a moderate inverse relationship between ability to recognize facial expressions of emotions in child faces and self-reported acts of physical violence. With respect to specific facial expressions of emotions in child faces, a moderate inverse relationship was found between ability to recognize the expression, fearful, and self-reported acts of physical violence. A moderate positive relationship was found between ability to recognized the expression, fearful, in child faces, and ability to empathize with the emotional experiences of others. Strong and moderate links were found between the negative expressions, fearful and sad, and angry and sad, respectively. Additionally, a strong inverse relationship was found between ability to emphathize with the emotional experiences of others and self-reported acts of delinquent behavior. Lastly, a strong positive relationship was found between covert and overt self reported acts of delinquent behavior. Results from this exploratory investigation suggest a link between facial affect recognition, empathy, and delinquency. Findings have important implications for educators and counselors who work with youth offenders within probation placement facilities. PMID- 16268137 TI - Gay adolescents and suicide: understanding the association. AB - Suicide among gay adolescents is a major public health concern, but it is still under-emphasized within mainstream healthcare. This article brings together information garnered from past studies, articles, and experiences to create a more in-depth understanding of why gay adolescents are at a higher risk for suicide. By creating a better understanding, this article attempts to push forward the significance of this issue and encourages physicians to take a more active role in suicide prevention. The article initially explores why issues involving homosexuality are seldom discussed and provides evidence for an increased risk in suicide among gay adolescents. By addressing major psychosocial issues faced by gay adolescents, it spends the majority of time facilitating an understanding of the increased risk. In this article "gay" refers to homosexual males and females, and bisexuals unless otherwise indicated. PMID- 16268138 TI - Acculturation models of immigrant Soviet adolescents in Israel. AB - This study probed acculturation in Israel of immigrant students from the former U.S.S.R. from the perception of the host society and the students. The finding from a questionnaire distributed to Israeli-born and immigrant students indicated that most but not all of the Israel-born students applied an assimilatory model while the immigrants were split between those who accepted the Israeli students and those who resisted them. Thus, ethnic relation typologies and critical sociology of education concepts can explain only part of the immigrants' attitudes. It was found that both the host society and the immigrants have different attitudes toward the culture of the other, and thus use different acculturation strategies. PMID- 16268139 TI - An investigation of Taiwanese early adolescents' views about the nature of science. AB - This study developed a Pupils' Nature of Science Scale, including the subscales of the invented and changing nature of science, the role of social negotiation on science, and cultural context on science, to assess early adolescents' views about the nature of science. More than 6,000 fifth and sixth graders in Taiwan responded to the Scale. The study revealed that the adolescents had quite different perspectives toward different subscales of the nature of science. Moreover, male adolescents tended to express more constructivist-oriented views toward the nature of science than did their female counterparts. The adolescents of different grades and races also displayed varying views toward the nature of science. PMID- 16268140 TI - Relational factors of vulnerability and protection for adolescent pregnancy: a cross-sectional comparative study of Portuguese pregnant and nonpregnant adolescents of low socioeconomic status. AB - This study explores multiple relational contexts that promote vulnerability and protection against early pregnancy in a potential risk group of Portuguese adolescents. A comparative analysis was made between two groups of female adolescents of low socioeconomic status: pregnant adolescents (n=57) and adolescents without a history of pregnancy (n=81). Results suggest that several variables belonging to different contexts-family and school and peer relations- are important in the characterization of the two groups. Lower levels of mother's overprotection and father's emotional support, presence of early pregnancy in adolescent's mother, lower level of emotional proximity to peer relations, and higher number of school failures are significantly associated with adolescent pregnancy. PMID- 16268141 TI - Comment on "The long-term fate of polychlorinated biphenyls in San Francisco Bay, (USA)". PMID- 16268142 TI - Correlation analyses for bimolecular nucleophilic substitution reactions of chloroacetanilide herbicides and their structural analogs with environmentally relevant nucleophiles. AB - Second-order rate constants (kNuc) for aqueous-phase bimolecular nucleophilic substitution (SN2) reactions of a range of anionic nucleophiles with alachlor, propachlor, and two analogs of propachlor (a thioacetanilide and a beta-anilide) were fit to the Swain-Scott and Edwards models. Correlations of literature kNuc values for analogous reactions of methyl chloride and methyl benzenesulfonate were included for comparison. The Swain-Scott correlation yielded poor to fair results for chloroacetanilides and their analogs, with adjusted (adj) r2 values ranging from 0.67 to 0.89, which are comparable to correlations for CH3Cl and CH3OSO2Ph (r2 (adj) = 0.80 and 0.85, respectively). Both the one- and two parameter Edwards models yielded improved correlations for the majority of substrates. A pronounced dependence on the substrate polarizability (alpha) factor generally was observed for the Edwards model, with a negligible dependence on the substrate basicity (beta) factor. Substrate polarizability factors for the one-parameter Edwards model were significantly larger for alachlor, propachlor, and the beta-anilide analog of propachlor than for methyl chloride and methyl benzenesulfonate. This indicates that the chloroacetanilides are activated toward SN2 reactions with highly reactive nucleophiles (e.g., HS-, PhS-, and Sn(2-)) relative to other saturated carbon substrates. At best, the Swain-Scott and Edwards models only furnish order-of-magnitude predictions of kNuc for the substrates investigated. PMID- 16268143 TI - Predicting sediment metal toxicity using a sediment biotic ligand model: methodology and initial application. AB - An extension of the simultaneously extracted metals/acid-volatile sulfide (SEM/AVS) procedure is presented that predicts the acute and chronic sediment metals effects concentrations. A biotic ligand model (BLM) and a pore water sediment partitioning model are used to predict the sediment concentration that is in equilibrium with the biotic ligand effects concentration. This initial application considers only partitioning to sediment particulate organic carbon. This procedure bypasses the need to compute the details of the pore-water chemistry. Remarkably, the median lethal concentration on a sediment organic carbon (OC)-normalized basis, SEM*(x,OC), is essentially unchanged over a wide range of concentrations of pore-water hardness, salinity, dissolved organic carbon, and any other complexing or competing ligands. Only the pore-water pH is important. Both acute and chronic exposures in fresh- and saltwater sediments are compared to predictions for cadmium (Cd), copper (Cu), nickel (Ni), lead (Pb), and zinc (Zn) based on the Daphnia magna BLM. The SEM*(x,OC) concentrations are similar for all the metals except cadmium. For pH = 8, the approximate values (micromol/gOC) are Cd-SEM*(xOC) approximately equal to 100, Cu-SEM*(x,OC) approximately equal to 900, Ni-SEMoc approximately equal to 1,100, Zn-SEM*(x,OC) approximately equal to 1,400, and Pb-SEM*(x,OC) approximately equal to 2,700. This similarity is the explanation for an empirically observed dose-response relationship between SEM and acute and chronic effects concentrations that had been observed previously. This initial application clearly demonstrates that BLMs can be used to predict toxic sediment concentrations without modeling the pore water chemistry. PMID- 16268144 TI - Evaluation of microbial inoculation and vegetation to enhance the dissipation of atrazine and metolachlor in soil. AB - Four greenhouse studies were conducted to evaluate the effects of native prairie grasses and two pesticide-degrading bacteria to remediate atrazine and metolachlor in soils from agricultural dealerships (Alpha site soil, northwest Iowa, USA; Bravo site soil, central Iowa, USA). The Alpha soil contained a low population of atrazine-degrading microorganisms relative to the Bravo soil. Each soil freshly treated with atrazine or metolachlor was aged for a short or long period of time, respectively. An atrazine-degrading bacterium, Agrobacterium radiobacter strain J14a; a metolachlor-degrading bacterium, Pseudomonas fluorescens strain UA5-40; and a mixture of three native prairie grasses-big bluestem (Andropogon gerardii Vitman), yellow Indian grass (Sorghastrum nutans [L.] Nash), and switchgrass (Panicum virgatum L.)-were added to the soils after the soils were aged for long periods of time. The soils aged for short periods of time were treated with J14a, the prairie grasses, or both after aging. The J14a and the grasses significantly reduced the concentration of atrazine in Alpha soil when the soil was aged for a short period of time. However, these treatments had no statistically significant effect when the soil was aged for a long period of time or on atrazine in Bravo soil. Inoculation with UA5-40 did not enhance metolachlor dissipation in either soil, but vegetation did increase metolachlor dissipation. Our results indicate that the dissipation of atrazine by J14a is affected by the presence of indigenous atrazine-mineralizing microorganisms and probably by the bioavailability of atrazine in the soil. PMID- 16268145 TI - Electron paramagnetic resonance analysis of the distribution of a hydrophobic spin probe in suspensions of humic acids, hectorite, and aluminum hydroxide humate-hectorite complexes. AB - Until recently, there were no techniques capable of direct observation of the microscale locations where nonpolar organic compounds accumulate when associated with natural geosorbents. The ability of electron paramagnetic resonance (EPR) spectroscopy to monitor and elucidate directly the different molecular-scale environments of paramagnetic spin probes has been demonstrated lately in model soils, yet it remains untested in complex systems. In this general context, the present investigation was aimed at assessing the extent to which EPR could be used to monitor the sorption of 4-hydroxy-2,2,6,6-tetramethyl-piperidinyloxy benzoate (TEMPO benzoate), a hydrophobic spin probe, on a smectite (hectorite), two humic acids, and their complexes in the presence or absence of aluminum hydroxide. Results demonstrate that EPR is able to monitor easily adsorption on these sorbents in batch-style experiments. Distribution coefficient (Kd) values of 455.4 and 483.1 ml/g were found for the adsorption of TEMPO benzoate on hectorite-humic acids complexes, compared to respective Kd values of 46 and 147 ml/g predicted solely on the basis of the mass of humic acids present in the complexes. These observations confirm the significant role of hectorite for the sorption of hydrophobic compounds, together with humic acids, contrary to common belief that emphasizes the almost exclusive sorptive role of organic matter. In addition, for the first time, EPR is able to provide evidence that hydrophobic molecules in the presence of geosorbents can segregate in multimolecular clusters that are in equilibrium with aqueous probe concentrations below the probe's solubility threshold. Possible consequences of this clustering process in terms of the fate and transport of hydrophobic compounds in subsurface environments are discussed. PMID- 16268146 TI - Effect-directed analysis of mutagens and ethoxyresorufin-o-deethylase inducers in aquatic sediments. AB - Sediment extracts from a creek in the Neckar river basin (Germany), which received the discharge of treated hospital wastewater, were found to exhibit strong aromatic hydrocarbon (Ah) receptor-mediated effects in a rainbow trout liver cell line (RTL-WI) as well as high mutagenicity in the Salmonella/microsome assay after fractionation. The crude extract did not exhibit a clear mutagenic response. Apparently, cleanup or fractionation before mutagenicity testing is necessary to minimize the risk of false-negative results. Effect-directed fractionation and analysis were applied to characterize and identify the toxicants that cause these effects. Major ethoxyresorufin-O-deethylase induction potency and mutagenicity were detected in different polyaromatic fractions, indicating different sets of toxicants that induce metabolic activation and mutagenicity. Dioxin-like halogenated aromatic hydrocarbons, including polychlorinated biphenyls, naphthalenes, dibenzo-p-dioxins and furans, and priority polycyclic aromatic hydrocarbons, contributed to Ah receptor-mediated activity only to a minor extent. Benzo[a]pyrene, benzo[a]fluoranthene, and perylene could be confirmed as important contributors to mutagenicity. The nonpriority pollutants 11H-indeno[2,1,7-cde]pyrene, a methylbenzo[e]pyrene, and a methylperylene were tentatively identified as major components, representing 82% of the peak area of a highly mutagenic fraction of the sediment extract. This suggests that hazard and risk assessment of complex environmental mixtures should make increasing attempts to identify and consider hazardous key pollutants rather than focusing on a priori-selected key pollutants alone. PMID- 16268147 TI - Low hepatic ethoxyresorufin-O-deethylase activity correlates with high organochlorine concentrations in Atlantic tomcod from the Canadian east coast. AB - A suppression of hepatic ethoxyresorufin-O-deethylase (EROD) activity was reported recently in large-sized Atlantic tomcod (Microgadus tomcod) from the St. Lawrence Estuary (SLE; QC, Canada), possibly related to chronic exposure to persistent contaminants and/or to emaciation. In the present study, hepatic concentrations of organochlorine contaminants and biological responses were measured in female tomcods from three estuaries located on the Canadian east coast: The SLE, the Miramichi (ME), and the Richibucto (RE) Rivers Estuaries (NB, Canada). Tomcods from the SLE had higher hepatic concentrations of organochlorine contaminants than tomcods from the ME and RE. For example, concentrations of polychlorinated biphenyls (PCBs, lipid wt) were 2.5 to 4 times higher, and concentrations of mirex and chlordanes were 6 times higher, in tomcods from the SLE than in tomcods from the other sites. Concentrations of polycyclic aromatic hydrocarbons (PAHs) metabolites in the bile did not differ among sites. The pattern of biological responses differed markedly between the SLE and the two other sites. Tomcods from the SLE had 1.5 times higher concentrations of DNA adducts and 2 times higher rates of hepatocellular proliferation, but 20 times lower hepatic EROD activity, than tomcods from the ME and RE. Lipid content was not correlated with EROD activity, indicating that low hepatic lipid content alone does not cause suppression of EROD activity in Atlantic tomcod. In contrast, for the three sites combined, EROD activity decreased as concentrations of PCBs increased. Within sites, hepatic PCB concentrations increased as lipid content decreased. This study supports the hypothesis that low EROD activity in SLE tomcods is related to chronic exposure to organochlorine contaminants. PMID- 16268148 TI - Prediction and experimental validation of acute toxicity of beta-blockers in Ceriodaphnia dubia. AB - Acute toxicity of beta-adrenoceptor blockers (beta-blockers) was studied with beta-blockers as single compounds or in mixture using the standardized acute 2-d Ceriodaphnia dubia immobility test. The tested compounds were selected according to their selectivity for the beta1-adrenoceptor, with three beta1-selective blockers (acebutolol, atenolol, and metoprolol) and three non-beta1-selective blockers (nadolol, oxprenolol, and propranolol). The acute toxicity (median effective concentration) of the six single compounds ranged from 1.4 mg/L for propranolol to 163 mg/L for nadolol. According to European Union directive 93/67EEC, these values range from toxic for aquatic organisms to nonclassified. The more toxic compounds, propranolol and oxprenolol, are both characterized by a membrane-stabilizing activity, a strong affinity for the beta1-adrenoceptor, and a high octanol-water partition coefficient (log Kow). The property of beta receptor selectivity seems not to be involved in the observed acute toxicity of the single compounds for C. dubia. Nevertheless, the toxicity of the selected compounds in mixture can be defined according to the beta1-selectivity. Two main joint effects have been detected: An independent action for the beta1-selective blockers, and an additive effect when either the nonselective beta1-selective blockers or the six compounds are tested together. The concentration addition model seems to be appropriate, providing a reasonable worst-case estimation of beta-blocker mixture toxicity for regulatory purposes. PMID- 16268149 TI - Functional and structural impact of linuron on a freshwater community of primary producers: the use of immobilized algae. AB - An approach in determining ecosystem integrity and stress on ecosystem level is to assess processes within ecosystems. The aim of the present study was to evaluate the potential use of an in situ assay with immobilized Chlorella vulgaris as an indicator of effects on ecosystem functioning with regard to primary production. The herbicide linuron, applied in concentrations of 20, 60, and 180 microg linuron/L, was used to induce direct effects on primary producers. Direct and indirect changes in structure and function within outdoor model ecosystems of 3 m3 were monitored. The intermediate and highest concentration of linuron caused a decline in growth of the macrophyte Elodea sp., resulting in a significant increase of concentrations of nutrients. The increase in concentrations of nutrients caused an indirectly stimulated growth of immobilized C. vulgaris at the intermediate concentration, whereas similar concentrations of nutrients, at the highest treatment, did not stimulate Chlorella growth. It appeared that the direct effect of linuron on C. vulgaris growth was masked by nutrient availability at the intermediate but not at the highest linuron concentration. The observed immobilized algal growth was an accumulated effect of toxic and trophic pressures within the ecosystem, resulting in an integrative endpoint to detect actual impairment of ecosystem function. PMID- 16268150 TI - New organochlorine contaminants and metabolites in plasma and eggs of glaucous gulls (Larus hyperboreus) from the Norwegian Arctic. AB - The present study investigated new or lesser-studied and legacy organochlorine (OC) contaminants and metabolites in plasma and eggs of glaucous gulls (Larus hyperboreus) collected from major breeding colonies on Bear Island in the Norwegian Arctic. Hexachlorobutadiene was below the method limit of detection (< 0.07 ng/g lipid wt) in all samples. The sum (sum) of 20 chlorobornane congener concentrations ranged between 294 and 986 ng/g lipid weight and 104 and 1,121 ng/g lipid weight in plasma and eggs, respectively, whereas those of sum 20polychlorinated naphthalene ranged between 1.34 and 126 ng/g lipid weight in plasma and 1.82 and 162 ng/g lipid weight in eggs. Bis(4-chlorophenyl) sulfone concentrations ranged between 5.24 and 143 ng/ g lipid weight plasma, which is the first report of this contaminant in arctic biota north of Sweden. Based on 2,3,7,8-tetrachlorodibenzo-p-dioxin toxic equivalents (TEQs), partial dioxin-like toxicity varied between 3.04 and 20.8 ng TEQ/g lipid weight in plasma and 0.94 and 46.5 ng TEQ/g lipid weight in eggs, and largely was due to concentrations of non-ortho polychlorinated biphenyls (PCBs), with a very minor contribution from mono-ortho PCBs and polychlorinated naphthalenes. The major aryl sulfone metabolite in plasma was an unidentified hexachlorinated MeSO2-PCB congener (range: 13.5-551 ng/g lipid wt), whereas the pentachlorinated congeners 3'- and 4'-MeSO2-CB101 (range: 4.49-38.1 ng/g lipid wt) dominated in eggs. The predominant halogenated phenolic compound (HPC) in plasma was consistently the PCB metabolite 4-OH-CB187 (range: 0.29-17.5 ng/g wet wt), whereas in eggs, detectable HPCs were at very low and transient concentrations. As part of a complex profile of contaminant exposure, these chemical classes and metabolites may be contributing factors to enhance physiological stress in breeding glaucous gulls. PMID- 16268151 TI - Effects of long-chain hydrocarbon-polluted sediment on freshwater macroinvertebrates. AB - High-molecular weight (> C16) hydrocarbons (HMWHs) are common pollutants in sediments of freshwater systems, particularly urban water bodies. No sediment quality guidelines exist for total hydrocarbons; more emphasis is placed on polyaromatic hydrocarbons, the most toxic component of hydrocarbons. A field based microcosm experiment was conducted to determine whether unpolluted sediments spiked with synthetic motor oil impair freshwater macroinvertebrate assemblages. Total petroleum hydrocarbon (TPH) concentrations of 860 mg/kg dry weight significantly increased the abundance of Polypedilum vespertinus and Cricotopus albitarsis and decreased the abundance of Paratanytarsus grimmii adults (all Chironomidae), whereas TPH concentrations ranging from 1,858 to 14,266 mg/kg produced a significant reduction in the total numbers of taxa and abundance, with significant declines in the abundance of nine chironomid taxa. About 28% of water bodies surveyed in urban Melbourne, Australia, had TPH concentrations in sediments likely to cause ecological impairment, and about 14% of the water bodies surveyed are likely to have reduced species richness and abundance. Therefore, HMWHs can be a significant pollutant in urban water bodies. Freshwater sediment quality guidelines should be developed for this ubiquitous urban pollutant. PMID- 16268152 TI - Weathering and aging of 2,4,6-trinitrotoluene in soil increases toxicity to potworm Enchytraeus crypticus. AB - Energetic materials are employed in a wide range of commercial and military activities and often are released into the environment. Scientifically based ecological soil-screening levels (Eco-SSLs) are needed to identify contaminant explosive levels in soil that present an acceptable ecological risk. Insufficient information for 2,4,6-trinitrotoluene (TNT) to generate Eco-SSLs for soil invertebrates necessitated toxicity testing. We adapted the standardized Enchytraeid Reproduction Test and selected Enchytraeus crypticus for these studies. Tests were conducted in Sassafras sandy loam soil, which supports relatively high bioavailability of TNT. Weathering and aging procedures for TNT amended to test soil were incorporated into the study design to produce toxicity data that better reflect the soil exposure conditions in the field compared with toxicity in freshly amended soils. This included exposing hydrated TNT-amended soils in open glass containers in the greenhouse to alternating wetting and drying cycles. Definitive tests showed that toxicity for E. crypticus adult survival and juvenile production was increased significantly in weathered and aged soil treatments compared with toxicity in freshly amended soil based on 95% confidence intervals. The median effect concentration and 20% effective concentration for reproduction were 98 and 77 mg/kg, respectively, for TNT freshly amended into soil and 48 and 37 mg/kg, respectively, for weathered and aged TNT soil treatments. These findings of increased toxicity to E. crypticus in weathered and aged TNT soil treatments compared with exposures in freshly amended soils show that future investigations should include a weathering and aging component to generate toxicity data that provide more complete information on ecotoxicological effects of energetic contaminants in soil. PMID- 16268153 TI - 1,3-dinitrobenzene toxicity in the least shrew, Cryptotis parva. AB - Shrews are abundant in most areas of toxic chemical contamination and bioaccumulate pollutants at much higher rates than sympatric rodent species. As a part of studies to provide information concerning the toxicity of 1,3 dinitrobenzene (DNB) in least shrews (Cryptotis parva), groups of 10 females and 10 males received DNB at 0 (control), 0.7, 2.9, 11.6, and 46.3 microl/L (equivalent mean daily dosage of 0, 0.26, 1.06, 4.26, and 17.0 mg/kg body wt in each sex) in their diet for 14 d. Leukocytosis present at the 0.26 mg/kg body weight/d dosage established the lowest-observed-adverse effect level (LOAEL). Adrenal enlargement was noted at the 1.06 mg/kg body weight/d level. Splenic enlargement and reductions in hematocrit and hemoglobin values occurred at the 4.26 mg/kg body weight/d treatment. Enlargements in the liver and heart and reductions in brown fat weight, granulocyte numbers, and alanine aminotransferase levels were present at high dose levels. Histopathologic examinations showed Kupffer's cell hemosiderosis and suggested testicular damage at the two highest tested doses but failed to confirm brain lesions. Least shrews do not follow standard scaling estimates for lifespan or metabolic rates. The LOAEL calculated from the standard terrestrial screening benchmark equation was higher than our findings, suggesting that these estimates must be viewed with caution. PMID- 16268155 TI - Effect of release herbicide on mortality, avoidance response, and growth of amphibian larvae in two forest wetlands. AB - Effects of Release herbicide (triclopyr butoxyethyl ester, [TBEE]) on mortality, avoidance response, and growth of larval amphibians (Rana clamitans, Rana pipiens) were investigated using in situ enclosures deployed in two forest wetlands in northern Ontario, Canada. Release was applied at nominal concentrations ranging from 0.26 to 7.68 mg TBEE acid equivalents (AE)/L. No significant deleterious effects of this herbicide on larval growth were detected. However, concentration-dependent mortality and abnormal avoidance response were observed. Most mortality occurred within 96 h following treatment. Median lethal concentration (LC50) values for each species and experimental site ranged from 2.79 to 3.29 mg AE/L, while median effective concentration (EC50) values (abnormal avoidance response) ranged from 1.67 to 3.84 mg AE/L. The LC10 and EC10 endpoints approximated aqueous concentrations (0.59 mg AE/L) expected under direct aerial overspray scenarios, indicating a potential risk of impacts for a small proportion of native amphibian larvae. However, given the low frequency and limited use of this herbicide formulation in Canadian forestry, these risks are considered negligible. Changes in usage patterns would require concurrent chemical and biological monitoring of operational spray programs to accurately quantify the probability and magnitude of real-world exposures and to relate these exposure levels to concentration-response relationships including those described in this study. PMID- 16268154 TI - Effects of the polycyclic aromatic hydrocarbon heterocycles, carbazole and dibenzothiophene, on in vivo and in vitro CYP1A activity and polycyclic aromatic hydrocarbon-derived embryonic deformities. AB - Heterocyclic derivatives of polycyclic aromatic hydrocarbons (PAHs) are often significant components of environmental contaminant mixtures; however, their contribution to the toxicity of these mixtures is not well characterized. These heterocycles commonly co-occur in PAH mixtures, which contain agonists for the aryl hydrocarbon receptor (AHR). Our goal for these studies was to explore the effects of two PAH heterocycles, carbazole (CB) and dibenzothiophene (DBT), alone and in combination with a PAH-type agonist for the AHR (beta-naphthoflavone [BNF]) on AHR-mediated cytochrome P4501A (CYP1A) activity and on fish embryotoxicity. Embryos of Fundulus heteroclitus were exposed to CB or DBT, with and without coexposure to BNE Carbazole alone slightly induced, whereas DBT alone slightly reduced, in ovo CYP1A-mediated ethoxyresorufin-O-deethylase (EROD) activity compared to control values. However, exposure to CB or DBT reduced in ovo EROD activity in embryos coexposed to BNE Carbazole and DBT were characterized in vitro as noncompetitive CYP1A inhibitors. Carbazole and DBT enhanced the embryotoxicity of BNF, although neither compound was embryotoxic by itself. The co-occurrence of CB and DBT with PAH-type AHR inducers in contaminated ecosystems may increase the toxicity of PAH-type AHR agonists in these settings and may need to be considered when estimating the embryotoxicity of PAH mixtures. PMID- 16268156 TI - Sensitivity of an indigenous amphipod (Corophium colo) to chemical contaminants in laboratory toxicity tests conducted with sediments from Sydney Harbor, Australia, and vicinity. AB - Laboratory survival tests were conducted with an indigenous infaunal amphipod, Corophium colo, on 103 sediment samples from Sydney Harbor (NSW, Australia) and vicinity, containing a wide range of chemicals and concentrations. The present study describes the sensitivity of C. colo to the sediments and compares the results to data for North American amphipods (Rhepoxynius abronius and Ampelisca abdita) previously used to establish and validate sediment-quality guidelines (SQGs). The incidence of toxicity increased with increasing contamination, as indicated by increasing numbers of SQGs exceeded and increasing mean SQG quotients. The incidence of highly toxic results (p < 0.05 and mean amphipod survival of < 80% that of controls) for highly contaminated samples was approximately half (28-40%) that of a large U.S. database (74%). The incidence of highly toxic responses for samples with intermediate levels of contamination also was lower in the present study (5-13%) compared to the results in large U.S. studies (approximately 30-50%). Corophium colo reburial tests showed greater sensitivity compared to survival tests, with a maximum incidence of statistically significant responses in moderately contaminated sediments of 70%. The present study showed that adult Corophium organisms are suitable for testing lethal responses in highly contaminated sediments (i.e., with mean effects range-median quotients of >1.5). Reburial results provide additional sensitivity. PMID- 16268157 TI - Analysis of estrogenic effects by quantification of green fluorescent protein in juvenile fish of a transgenic medaka. AB - TheChgH-GFP strain of the teleost medaka contains a regulatory region of the estrogen-responsive choriogenin H (chgH) gene fussed to the green fluorescent protein (GFP) gene. The strain was developed for the identification of environmental estrogens by noninvasive analysis of GFP fluorescence. In the present study, a quantification method for GFP by image analysis was establishedand applied to the analysis of time- and concentration-dependent GFP fluorescence in juvenile fish. Concentration-response analyses were performed with fish exposed for 14 d to 17beta-estradiol (0.37-367 pM), genistein (0.37-367 nM), or p-nonylphenol (0.367-1,835 nM). By means of image analysis, it was shown that ChgH-GFP was induced at 183.5 pM or greater 17beta-estradiol. Time-course and recovery experiments indicated a strong accumulation of GFP in the liver. Results of reverse transcriptase-polymerase chain reaction analysis of choriogenin H and vitellogenin demonstrated induction of gene expression for the same rangeof concentrations as that for GFP analysis. Neither expression of these genes nor GFP fluorescence was induced by genistein and p-nonylphenol. Although the ChgH-GFP strain failed to detect these weakly estrogenic compounds, the simplicity of the GFP quantification during early life stages of fish offers promising possibilities for further developments of transgenic strains using different target regulatory sequences. PMID- 16268158 TI - Persistent organic pollutants in the diet of harbor seals (Phoca vitulina) inhabiting Puget Sound, Washington (USA), and the Strait of Georgia, British Columbia (Canada): a food basket approach. AB - Harbor seals (Phoca vitulina) inhabiting Puget Sound (WA, USA) recently were found to be seven times more contaminated with polychlorinated biphenyls (PCBs) than those inhabiting the adjacent Strait of Georgia (BC, Canada). We carried out a food basket approach to approximate realistic dietary exposures of both new (e.g., polybrominated diphenyl ethers [PBDEs]) and legacy (e.g., dichlorodiphenyltrichloroethane [DDT]) persistent organic pollutants (POPs) for these harbor seals. Food basket homogenates, each consisting of over 200 individual prey items, were constructed using documented dietary preferences for harbor seals in these basins, and analyzed for organochlorine pesticides, flame retardants, and other persistent contaminants. Concentration rankings for the major contaminant classes in the Puget Sound food basket were sigmaPCBs > sigmaPBDEs > sigmaDDT, and for the Strait of Georgia food basket were sigmaPCBs > sigmaDDT > sigmaPBDEs, highlighting the emergence of PBDEs as a significant concern in the regional environment. Consistent with observations in harbor seals, PCB concentrations in the Puget Sound food basket were seven times higher than in its Strait of Georgia counterpart. Based on our food basket results, the estimated daily intake of sigmaPCB toxic equivalents to dioxin by Puget Sound harbor seals exceeds some wildlife consumption guidelines for PCBs. Our results indicate that both legacy and new POPs present a health risk to these marine mammals. PMID- 16268159 TI - Potential ecotoxic effects of polychlorinated biphenyls on Xenopus laevis. AB - We examined potential ecotoxic effects of polychlorinated biphenyl (PCB)3, PCB5, Aroclor 1254, and Aroclor 1242 on Xenopus laevis. Tadpoles were exposed to PCBs from stage 46/47 (system of Nieuwkoop and Faber) to the completion of metamorphosis. We demonstrated, to our knowledge for the first time, forelimb malformations caused by PCBs (malformation rate, > 70%). The malformed forelimbs were fixed in the adduction-backward rotation position and could not move. Therefore, malformed male frogs were destined to have no offspring, because they could not grasp the females with their forelimbs to mate. Alcian blue-alizarin red double-staining indicated that the forelimb malformation resulted from the shoulder abnormality. Compared with the normal shoulder joint, the proximal humerus with the humerus inter-rotated 90 degrees in the abnormal shoulder joint. Moreover, testes from more than a third of male frogs with exposed to PCBs exhibited feminization to different degrees at gross morphology and histology, with fewer or abnormal spermatogonia and oocytes. Gonadal abnormalities would lead directly to reproductive dysfunction and population decline. These results suggest that PCBs have potentially ecotoxic effects on amphibian populations. We infer that PCBs could play roles in amphibian malformations and population declines, at least at sites that are polluted heavily with PCBs. PMID- 16268160 TI - Survival and reproduction of enchytraeid worms, Oligochaeta, in different soil types amended with energetic cyclic nitramines. AB - Hexanitrohexaazaisowurtzitane (CL-20), a new polycyclic polynitramine, has the same functional nitramine groups (N-NO2) as the widely used energetic chemicals hexahydro-1,3,5-trinitro-1,3,5-triazacyclohexane (royal demolition explosive [RDX]) and octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine (high-melting explosive [HMX]). Potential impacts of CL-20 as an emerging contaminant must be assessed before its use. The effects of CL-20, RDX, or HMX on adult survival and juvenile production by potworms Enchytraeus albidus and Enchytraeus crypticus were studied in three soil types, including Sassafras sandy loam (1.2% organic matter [OM], 11% clay, pH 5.5), an agricultural soil (42% OM, 1% clay, pH 8.2), and a composite agricultural-forest soil (23% OM, 2% clay, pH 7.9) by using ISO method 16387 (International Standard Organization, Geneva, Switzerland). Results showed that CL-20 was toxic to E. crypticus with median lethal concentration values for adult survival ranging from 0.1 to 0.7 mg/kg dry mass (DM) when using the three tested soils. In addition, CL-20 adversely affected juvenile production by both species in all soils tested, with median effective concentration (EC50) values ranging from 0.08 to 0.62 mg/kg DM. Enchytraeus crypticus and E. albidus were similarly sensitive to CL-20 exposure in the composite agricultural-forest soil, which supported reproduction by both species and enabled comparisons. Correlation analysis showed weak or no relationship overall among the soil properties and reproduction toxicity endpoints. Neither RDX nor HMX affected (p > 0.05) adult survival of either species below 658 and 918 mg/kg DM, respectively, indicating that CL-20 is more toxic to enchytraeids than RDX or HMX. Examination of data shows that CL-20 should be considered as a potential reproductive toxicant to soil invertebrates, and that safeguards should be considered to minimize the potential for release of CL-20 into the environment. PMID- 16268161 TI - Effects of polychlorinated biphenyls on the schooling behavior of Japanese medaka (Oryzias latipes). AB - The effects of polychlorinated biphenyls (PCBs; Kanechlor 400) on the schooling behavior of Japanese medaka (Oryzias latipes) were tested after feeding with various concentrations of PCBs (0, 1, 5, 25, and 125 microg/g). To test schooling, three PCB-exposed medaka and three untreated fish were placed in one chamber, and their swimming trajectories were recorded. Swimming velocity, turning angle, and nearest-neighbor distance (NND) were analyzed individually, and the polarization and expanse of each schooling group were analyzed. Fractal dimension analysis was performed for trajectory, swimming velocity, and turning angle. Six behaviors (school formed by six swimming medaka, school formed by six stationary medaka, school formed by three to five swimming medaka, school formed by three to five stationary medaka, swimming without schooling, and stationary without schooling) were quantified with time. Behavioral parameters were the same for PCB-exposed and unexposed medaka in the same schooling group. Swimming velocity decreased in a dose-dependent manner, and NND in the group exposed to 25 microg/g of PCBs was significantly longer (126%) than that in the controls. The fractal dimensions of trajectory and turning angle increased significantly in the highest PCB-exposure group, indicating that schools containing PCB-exposed individuals frequently changed direction. The PCBs shortened the time of school formed by six swimming fish, which was 23.7% of the control value in the group exposed to 25 microg/g of PCBs. The frequency of collisions between individuals was significantly greater in the highest-exposure group (31.3) than in the controls (13.3). Thus, PCB exposure influences the behavior of unexposed fish in the same school, consequently affecting schooling behavior. PMID- 16268162 TI - Effects of maternally transferred organochlorine contaminants on early life survival in a freshwater fish. AB - Laboratory research has shown that female fish can pass toxic organochlorines (OCs) from their bodies to their eggs, killing their offspring if sufficient quantities are transferred. We conducted a controlled incubation study using gametes from a wild, OC-contaminated walleye (Sander vitreus) population (Bay of Quinte, Lake Ontario, Canada) in order to assess among-female variation in offspring early life survival in relation to ova concentrations of planar OCs (polychlorinated dibenzo-p-dioxins and furans and planar polychlorinated biphenyls) and a suite of other maternal and ova characteristics. Equal volumes of ova from each female were fertilized, pooled, and incubated together as an experimental cohort. Relative survival of each female's offspring was estimated as the proportion of surviving larvae (at approximately 5 d posthatch) that she contributed to the cohort as determined by microsatellite DNA parentage assignment. Total planar OC concentration (expressed as toxic equivalency of 2,3,7,8-tetrachlorodibenzo-p-dioxin) of ova was positively related to maternal age and size and to ova lipid content. However, early life survival did not decline with increasing ova planar OC concentrations. Similarly, we observed no significant relationships between early life survival and ova thiamine content, ova fatty acid composition, or maternal age or size. Early life survival was more strongly correlated with date of spawn collection, thyroid hormone status of the ova, and ovum size. Maternally transferred planar OCs do not appear to negatively influence female reproductive success in this walleye population. PMID- 16268164 TI - Validation of otolith growth rate analysis using cadmium-exposed larval topsmelt (Atherinops affinis). AB - We applied otolith growth rate analysis to an investigation of cadmium (Cd) exposed larval topsmelt (Atherinops affinis) to determine if growth rate was a more sensitive measure than somatic growth (body wt or length). Topsmelt otoliths, calcareous concretions in the fish inner ear, formed daily increments, and otolith growth was proportional to somatic growth. Nine-day posthatch larval topsmelt were exposed to Cd (0-100 ppb) in seawater for 14 d and fed low or high ration levels in separate experiments. Whereas Cd impaired topsmelt growth and growth rates, the extent of growth reduction was dependent on the ration level. At high ration levels, otolith and somatic growth rates of fish exposed to Cd (50 and 100 ppb) were significantly reduced; however, no differences in final mean weight and only marginal differences in final mean length of Cd-exposed topsmelt were observed. At low ration levels, we detected reductions in both somatic growth as well as otolith and somatic growth rates of topsmelt exposed to Cd (50 and 100 ppb). Otolith growth rate analysis was more sensitive than growth measurements of Cd-exposed topsmelt, because it allowed the detection of small differences in growth rates even when differences in somatic growth were not observed. PMID- 16268163 TI - Comparison of Cd, Cu, and Zn toxic effects on four marine phytoplankton by pulse amplitude-modulated fluorometry. AB - The toxic effects of Cd, Cu, and Zn on four different marine phytoplankton, Dunaliella tertiolecta, Prorocentrum minimum, Synechococcus sp., and Thalassiosira weissflogii, were examined by comparing the cell-specific growth rate, pulse-amplitude-modulated (PAM) parameters (maximum photosystem II quantum yield phiM and operational quantum yield phi'M, chlorophyll a content, and cellular metal concentration, over a 96-h period. The calculated no-observed effect concentration (NOEC) based on both cell-specific growth rate and two PAM parameters (phiM and phi'M) were mostly identical. Thus, these PAM parameters and cell-specific growth rate were comparable in their sensitivities as the biomarkers for trace metal toxicity to marine phytoplankton. The cyanobacteria Synechococcus sp. was the most sensitive species among the four algal species tested because of its higher cell surface to volume ratio. The toxicity of the three tested metals followed the order of Cd > Cu > Zn based on the cellular metal concentration of the four algae at the NOEC. The cellular metal bioaccumulation followed the same Freundlich isotherm for each metal regardless of the algal species, indicating that the metal accumulation was a nonmetabolic process under high ambient metal concentrations and that the cell surface metal binding was comparable among the different species. For all the algae examined in our study, the bioaccumulation potentials of Cu and Zn were similar to each other, while the Cd bioaccumulation was much lower under environmentally realistic metal concentration. PMID- 16268165 TI - Effects of hydrogen sulfide to Vibrio fischeri, Scenedesmus vacuolatus, and Daphnia magna. AB - The effects of hydrogen sulfide (H2S) were tested in three ecotoxicological tests in order to evaluate its confounding potential in assessment of pore water and groundwater toxicity. The luminescent bacteria Vibrio fischeri, the water flea Daphnia magna, and the microalgae Scenedesmus vacuolatus often are part of a biotest battery. A new technique for the synthesis of hydrogen sulfide solutions of defined concentrations using an electrochemical generator instead of sodium sulfide solutions was used. Because hydrogen sulfide is volatile, the loss rate of H2S was studied over time to enable estimation of the mean test concentrations over the whole test duration. Loss rates were calculated to be 13 +/- 6% after 30 min, and 39 +/- 11% and 43 +/- 16% after a 24- and 48-h exposure time, respectively. Sensitivities of the test organisms in terms of median effective concentration (EC50), corrected for the above loss rates, varied from 0.28 to 0.0036 and 0.055 mM for the luminescent bacteria, the crustacea, and the algae, respectively. A species-sensitivity distribution using EC and mean lethal concentration literature data for marine and freshwater crustaceans and phytoplankton showed a medium sensitivity of the water flea D. magna, though the bacteria V. fischeri and the algae S. vacuolatus were among the least-sensitive group of organisms. This demonstrates that only the algae and the bacteria are easy to use in the assessment of toxicity of matrices with H2S concentrations above 0.06 mM. PMID- 16268166 TI - Toxicity, biotransformation, and mode of action of arsenic in two freshwater microalgae (Chlorella sp. and Monoraphidium arcuatum). AB - The toxicity of As(V) and As(III) to two axenic tropical freshwater microalgae, Chlorella sp. and Monoraphidium arcuatum, was determined using 72-h growth rate inhibition bioassays. Both organisms were tolerant to As(III) (72-h concentration to cause 50% inhibition of growth rate [IC50], of 25 and 15 mg As[III]/L, respectively). Chlorella sp. also was tolerant to As(V) with no effect on growth rate over 72 h at concentrations up to 0.8 mg/L (72-h IC50 of 25 mg As[V]/L). Monoraphidium arcuatum was more sensitive to As(V) (72-h IC50 of 0.25 mg As[V]/L). An increase in phosphate in the growth medium (0.15-1.5 mg PO4(3-)/L) decreased toxicity, i.e., the 72-h IC50 value for M. arcuatum increased from 0.25 mg As(V)/L to 4.5 mg As(V)/L, while extracellular As and intracellular As decreased, indicating competition between arsenate and phosphate for cellular uptake. Both microalgae reduced As(V) to As(III) in the cell, with further biological transformation to methylated species (monomethyl arsonic acid and dimethyl arsinic acid) and phosphate arsenoriboside. Less than 0.01% of added As(V) was incorporated into algal cells, suggesting that bioaccumulation and subsequent methylation was not the primary mode of detoxification. When exposed to As(V), both species reduced As(V) to As(III); however, only M. arcuatum excreted As(III) into solution. Intracellular arsenic reduction may be coupled to thiol oxidation in both species. Arsenic toxicity most likely was due to arsenite accumulation in the cell, when the ability to excrete and/or methylate arsenite was overwhelmed at high arsenic concentrations. Arsenite may bind to intracellular thiols, such as glutathione, potentially disrupting the ratio of reduced to oxidized glutathione and, consequently, inhibiting cell division. PMID- 16268167 TI - Sorption and degradation of estrogen-like-endocrine disrupting chemicals in soil. AB - Studies were undertaken to assess sorption of seven endocrine-disrupting chemicals (EDCs), namely, estrone (E1), 17beta-estradiol (E2), estriol (E3), 17alpha-ethynylestradiol (EE2), bisphenol A (BPA), 4-tert-octyl phenol (4-t-OP), and 4-n-nonyl phenol (4-n-NP) on four soils (from sandy to clay soil) with different physicochemical properties and biodegradation of five EDCs (BPA, E2, EE2, 4-t-OP, and 4-n-NP) in a loam soil associated with wastewater reuse. We also characterized the biotransformation of E2 to El in the loam soil under aerobic and anaerobic conditions. Sorption test using a batch equilibrium method demonstrated that alkylphenols (4-t-OP and 4-n-NP) had the strongest sorption onto soils, followed by estrogens (EE2, E2, El, E3) and BPA. This laboratory study showed that all five EDCs, including the degradation product El, were degraded rapidly in the soil within 7 d under aerobic conditions. However, under anaerobic conditions in the soil, little or no degradation of the five EDCs was noted except for E2, which showed slow degradation during the 70-d study. The calculated half-lives for E2 under anaerobic conditions were 24 d in the soil. Estradiol was found to be biotransformed to E1 under both aerobic and anaerobic conditions. The study suggested that the five EDCs as well as El, which may be present in reclaimed wastewater, would not persist in well-aerated soil. But these EDCs persisting in anaerobic soil may affect soil and groundwater quality and ecosystem. PMID- 16268168 TI - Comparison of different androgen bioassays in the screening for environmental (anti)androgenic activity. AB - Endocrine disruptors pose a growing threat to human and wildlife health. Validated test systems are required to study the mechanisms by which chemicals may interfere with the endocrine system. In order to identify compounds with (anti)androgenic activity, we used several in vitro bioassays, based on different androgen receptor (AR) functions, including AR transactivation and interaction between the amino-terminal domain and the ligand-binding domain. The AR activity assay, based on activation of the transcription of an androgen-responsive reporter gene in the presence of androgen, proved to excel in terms of high fold induction range and low minimal detection limit. The EC50 value, defined as the concentration that leads to half the maximal response and reflecting the potency of the synthetic androgen R1881 (methyltrienolone), was found to be 250 pM, consistent with the high affinity of this ligand to the human AR. A number of environmental samples were tested in the bioassay. The bioassay also could be used to detect antiandrogenic activity, because known AR-antagonists were able to inhibit R1881-mediated transactivation. PMID- 16268169 TI - Metal and oxygen uptake in the green mussel Perna viridis under different metabolic conditions. AB - Coupled respirometric and radiotracer techniques were applied to simultaneously measure the rates of oxygen and metal uptake in the green mussel Perna viridis. This was performed under different metabolic conditions by varying the ambient oxygen partial pressure (P(O2)), temperature, air exposure, and body size. When the mussels were tested under different hypoxic and anoxic conditions, Cd and Zn uptake decreased with decreasing P(O2), accompanied by a decrease in the ventilation activity of mussels. Significant reduction in metal uptake was observed at a P(O2) level of 3 kPa. Under anoxic conditions, the uptake of Cd and Zn was 1.6 to 2.7 times and 2.8 times, respectively, lower than those measured under normoxia. In contrast, both the absorption efficiencies of Cd and Zn and the oxygen extraction efficiency increased significantly with decreasing P(O2). There were significant correlations between the rates of Cd/Zn and O2 uptake by the mussels when quantified under various P(O2) levels. The uptake of Cd and Zn was temperature dependent and increased with temperature over a range of 15 to 30 degrees C. Significant correlations between the rates of Cd/Zn and O2 uptake were also found in the temperature experiments. With reimmersion of mussels after aerial exposure, the mussels experienced an apparent O2 debt. Metal uptake also increased within the first 15 min followed by gradual recovery to the control levels. Similarly, the quantified uptake rates of Zn were significantly correlated with the O2 uptake in experiments with different sizes of mussels. These results strongly suggest that Cd and Zn uptake are coupled with oxygen uptake in the mussels; thus, physiological processes need to be considered in studying metal accumulation. PMID- 16268170 TI - Complex mixture toxicity for single and multiple species: proposed methodologies. AB - Methods for the assessment of ecological risks associated with exposure to defined mixtures of toxicants are reviewed and formalized for single-species toxicity. Depending on the modes of action of toxicants in a mixture, these methods apply either the model for concentration additivity (CA) or the model for response additivity (RA). For complex mixtures, the present paper advocates the use of a new, two-step, mixed-model approach as a logical extension of model selection: Mixture toxicity for individual modes of action is evaluated with the CA model, and the toxicities of different modes of action are combined using the RA model. Using comparable mixture toxicity strategies in combination with the concept of species-sensitivity distributions, we develop a method to address and predict the risk for direct effects on the composition of species assemblages and biodiversity. The data needed for modeling can be obtained from existing databases, and lack of data can, in part, be addressed by the use of toxicity patterns in those databases. Both single- and multiple-species methods of mixture risk prediction are useful for risk management, because they allow ranking of polluted sites and affected species as well as identification of the most hazardous contaminants, at least in a comparative way. Validation of the proposed methods is feasible but currently limited because of a lack of appropriate data. PMID- 16268172 TI - An evaluation of logistic regression models for predicting amphipod toxicity from sediment chemistry. AB - An empirical screening level approach was developed to assess the probability of toxicity to benthic organisms associated with contaminated sediment exposure. The study was based on simple logistic regression models (LRMs) of matching sediment chemistry and toxicity data retrieved from a large database of field-collected sediment samples contaminated with multiple chemicals. Three decisions were made to simplify the application of LRMs to sediment samples contaminated with multiple chemicals. First, percent mortality information associated with each sediment sample was condensed into a dichotomous response (i.e., toxic or nontoxic). Second, each LRM assumed that toxicity was attributable to a single contaminant. Third, individual contaminants present at low concentrations were excluded from toxic sediment samples. Based on an analysis of the National Sediment Inventory database, the LRM approach classified 55% of nontoxic sediments as toxic (i.e., false-positives). Because this approach has been used to assess the probability of benthic toxicity as reported by the U.S. Environmental Protection Agency (U.S. EPA), the resultant estimates of potential toxicity convey a misleading impression of the increased hazard that sediments pose to the health of aquatic organisms at many sites in the United States. This could result in important resources needlessly being diverted from truly contaminated sites to evaluate and possibly remediate sediments at uncontaminated sites. PMID- 16268173 TI - Significance testing of synergistic/antagonistic, dose level-dependent, or dose ratio-dependent effects in mixture dose-response analysis. AB - In ecotoxicology, the state of the art for effect assessment of chemical mixtures is through multiple dose-response analysis of single compounds and their combinations. Investigating whether such data deviate from the reference models of concentration addition and/or independent action to identify overall synergism or antagonism is becoming routine. However, recent data show that more complex deviation patterns, such as dose ratio-dependent deviation and dose level dependent deviation, need to be addressed. For concentration addition, methods to detect such deviation patterns exist, but they are stand-alone methods developed separately in literature, and conclusions derived from these analyses are therefore difficult to compare. For independent action, hardly any methods to detect such deviations from this reference model exist. This paper describes how these well-established mixture toxicity principles have been incorporated in a coherent data analysis procedure enabling detection and quantification of dose level-and dose ratio-specific synergism or antagonism from both the concentration addition and the independent action models. Significance testing of which deviation pattern describes the data best is carried out through maximum likelihood analysis. This analysis procedure is demonstrated through various data sets, and its applicability and limitations in mixture research are discussed. PMID- 16268171 TI - Application of frog embryo teratogenesis assay-Xenopus to ecological risk assessment. AB - An expert workshop recently was convened to consider the frog embryo teratogenesis assay-Xenopus (FETAX) as a screening method for identifying the potential developmental toxicity of single chemicals and chemical mixtures. One recommendation from the workshop was that, in order to determine the utility of FETAX for ecological risk assessments, additional consideration of how the assay is conducted is necessary. In addition, a comparative evaluation would be useful of FETAX endpoints (i.e., survival, malformations, growth) versus each other, endpoints from aquatic toxicity tests using more commonly tested species of cladocerans and fish, and tests with other amphibian species. This review provides an evaluation and critique of the current FETAX protocol from two perspectives: Practical considerations relative to conducting the test and sensitivity of the assay (and associated endpoints) compared to tests with other species. Several aspects of the current standard protocol, including test temperature, diet, loading rates, and chemical exposure options, need to be modified to ensure that the assay is robust technically. Evaluation of FETAX data from the open literature indicates that growth is the most sensitive endpoint in the assay, followed by malformations and then survival; unfortunately, the growth endpoint often is not considered or reported in the assay. Comparison of FETAX data with acute toxicity data from tests with other amphibians or traditional aquatic test species indicates FETAX is relatively insensitive. This suggests that environmental risk assessments using acute hazard data from tests with traditional aquatic test species usually would be more protective of native amphibian species than risk assessments that use hazard data from FETAX. PMID- 16268174 TI - Registration of confocal scanning laser microscopy and quantitative backscattered electron images for the temporospatial quantification of mineralization density in 18-month old thoroughbred racehorse articular calcified cartilage. AB - Combined backscattered electron scanning electron microscopy (BSE SEM) and confocal scanning laser microscopy (CSLM) have been used to put tissue mineralization data into the context of soft tissue histology and fluorescent label information. Mineralization density (Dm) and linear accretion rate (LAR) are quantifiable parameters associated with mineralizing fronts within calcified tissues. Quantitative BSE (qBSE) may be used to determine Dm, while CSLM may be used to detect label fluorescence from which LAR is calculated. Eighteen-month old Thoroughbred horses received single calcein injections 19 and 8 days prior to euthanasia, labeling sites of active mineralization with fluorescent bands. Confocal scanning laser microscopy images of articular calcified cartilage (ACC) from distal third metacarpal condyles were registered to qBSE images of the same sites using an in-house program. ImageJ and Sync Windows enabled the simultaneous collection of LAR and Dm data. The repeatability of the registration and measurement protocols was determined. Dm profiles between calcein labels were explored for an association with time. Dm was 119.7 +/- 24.5 (mean +/- standard deviation) gray levels (where 0 = backscattering from monobrominated and 255 from monoiodinated dimethacrylate standards, respectively), while modal and maximum LAR were 0.45 and 3.45 microm/day, respectively. Coefficients of variation (CV) for Dm were 0.70 and 0.77% with and without repeat registration, respectively; CVs for LAR were 1.90 and 2.26% with and without repeat registration, respectively. No relationship was identified between Dm and time in the 11-day interlabel interval. Registration of CSLM to qBSE images is sufficiently repeatable for quantitative studies of equine ACC. PMID- 16268175 TI - X-ray spectrometry and spectrum image mapping at output count rates above 100 kHz with a silicon drift detector on a scanning electron microscope. AB - A third-generation silicon drift detector (SDD) in the form of a silicon multicathode detector (SMCD) was tested as an analytical x-ray spectrometer on a scanning electron microscope. Resolution, output count rate, and spectral quality were examined as a function of the detector peaking time from 8 micros to 250 ns and over a range of input count rate (dead time). The SDD-SMCD (50 mm2 active area) produced a resolution of 134 eV with a peaking time of 8 micros. The peak width and peak channel were nearly independent of the input count rate (at 8 micros peaking time, the peak width degradation was 0.003 eV/percent dead time and peak position change was -0.7 eV over the dead time range tested). Maximum output count rates as high as 280 kHz were obtained with a 500 ns peaking time (188 eV resolution) and 500 kHz with a 250 ns peaking time (217 eV resolution). X ray spectrum imaging was achieved with a pixel dwell time as short as 10 ms (with 1.3 ms overhead) in which a 2048 channel (10 eV/channel) spectrum with 2-byte intensity range was recorded at each pixel (scanned at 128 x 128). With a 220 kHz output count rate, a minor constituent of iron (present at a concentration of 0.04 mass fraction or 4 weight %) in an aluminum-nickel alloy could be readily detected in the x-ray maps derived from the x-ray spectrum image database accumulated in 185 s. PMID- 16268176 TI - Mineral content changes in bone associated with damage induced by the electron beam. AB - Energy-dispersive x-ray (EDX) spectroscopy and backscattered electron (BSE) imaging are finding increased use for determining mineral content in microscopic regions of bone. Electron beam bombardment, however, can damage the tissue, leading to erroneous interpretations of mineral content. We performed elemental (EDX) and mineral content (BSE) analyses on bone tissue in order to quantify observable deleterious effects in the context of (1) prolonged scanning time, (2) scan versus point (spot) mode, (3) low versus high magnification, and (4) embedding in poly-methylmethacrylate (PMMA). Undemineralized cortical bone specimens from adult human femora were examined in three groups: 200x embedded, 200x unembedded, and 1000x embedded. Coupled BSE/EDX analyses were conducted five consecutive times, with no location analyzed more than five times. Variation in the relative proportions of calcium (Ca), phosphorous (P), and carbon (C) were measured using EDX spectroscopy, and mineral content variations were inferred from changes in mean gray levels ("atomic number contrast") in BSE images captured at 20 keV. In point mode at 200x, the embedded specimens exhibited a significant increase in Ca by the second measurement (7.2%, p < 0.05); in scan mode, a small and statistically nonsignificant increase (1.0%) was seen by the second measurement. Changes in P were similar, although the increases were less. The apparent increases in Ca and P likely result from decreases in C: -3.2% (p < 0.05) in point mode and -0.3% in scan mode by the second measurement. Analysis of unembedded specimens showed similar results. In contrast to embedded specimens at 200x, 1000x data showed significantly larger variations in the proportions of Ca, P, and C by the second or third measurement in scan and point mode. At both magnifications, BSE image gray level values increased (suggesting increased mineral content) by the second measurement, with increases up to 23% in point mode. These results show that mineral content measurements can be reliable when using coupled BSE/EDX analyses in PMMA-embedded bone if lower magnifications are used in scan mode and if prolonged exposure to the electron beam is avoided. When point mode is used to analyze minute regions, adjustments in accelerating voltages and probe current may be required to minimize damage. PMID- 16268177 TI - Read-out of soft X-ray contact microscopy microradiographs by focused ion beam/scanning electron microscope. AB - A novel focused ion beam-based technique is presented for the read-out of microradiographs of Caenorhabditis elegans nematodes generated by soft x-ray contact microscopy (SXCM). In previous studies, the read-out was performed by atomic force microscopy (AFM), but in our work SXCM microradiographs were imaged by scanning ion microscopy (SIM) in a focused ion beam/scanning electron microscope (FIB/SEM). It allows an ad libitum selection of a sample region for gross morphologic to nanometric investigations, with a sequence of imaging and cutting. The FIB/SEM is less sensitive to height variation of the relief, and sectioning makes it possible to analyse the sample further. The SXCM can be coupled to SIM in a more efficient and faster way than to AFM. Scanning ion microscopy is the method of choice for the read-out of microradiographs of small multicellular organisms. PMID- 16268178 TI - Monte Carlo simulation of secondary electron and backscattered electron images in scanning electron microscopy for specimen with complex geometric structure. AB - A new Monte Carlo technique for the simulation of secondary electron (SE) and backscattered electron (BSE) of scanning electron microscopy (SEM) images for an inhomogeneous specimen with a complex geometric structure has been developed. The simulation is based on structure construction modeling with simple geometric structures, as well as on the ray-tracing technique for correction of electron flight-step-length sampling when an electron trajectory crosses the interface of the inhomogeneous structures. This correction is important for the simulation of nanoscale structures of a size comparable with or even less than the electron scattering mean free paths. The physical model for electron transport in solids combines the use of the Mott cross section for electron elastic scattering and a dielectric function approach for electron inelastic scattering, and the cascade SE production is also included. PMID- 16268179 TI - Say goodbye to the bench...microwave is here to stay! PMID- 16268180 TI - [Citizens' language]. PMID- 16268181 TI - Obesity risk factors in the ISAAC (International Study of Asthma and Allergies in Childhood) in Mexico City. AB - BACKGROUND: The International Study of Asthma and Allergies in Childhood (ISAAC) has promoted surveys in asthma and allergic diseases using standardized methodologies including validated questionnaires. Many items in the questionnaires have also been implied in the overweight and obesity etiology. OBJECTIVE: To describe the factors associated with obesity in subjects of 6-7 years and 13-14 years in the ISAAC survey in Mexico City. MATERIAL AND METHODS: Data were obtained from questionnaires of children participating in a phase 3b ISAAC survey. Logistic regression was used to determine the obesity risks factors. RESULTS: The factors related to obesity were weekly consumption of meat (+, positive relationship), vegetables, pasta, rice (+) and quartiles of birth weight (+) in boys of 6-7 years. Having suffered eczema at any time, weekly consumption of fruit, pasta, butter, nuts, potato (+), fast food (+), daily TV viewing (+) in girls of 6-7years. Having suffered eczema at any time, weekly consumption of pasta (+), butter, potato, weekly physical exercise in boys of 13 14 years; weekly consumption of pasta, margarine, milk, fast food (+), currently smoking in girls of 13-14 years. CONCLUSIONS: There were not common factor patterns for the different groups, birth weight, fast food, TV viewing and lack of exercise have been previously related to pediatric obesity. Asthma was not associated with a higher risk of obesity but medical history of eczema was associated with lower risk of obesity in the 6-7 years girls, and 13-14 years boys. The present study provides the bases for future epidemiological studies and gives some clues on possible public health actions. PMID- 16268182 TI - [Diminished apoptosis in nasal mucosa of patients with persistent allergic rhinitis]. AB - OBJECTIVE: To assess apoptosis frequency in the nasal mucosa of patients with perennial allergic rhinitis. PATIENTS AND METHODS: Eleven patients and five healthy subjects accepted to participate, from 10 to 16 years old. None of them had evidence of acute infectious disease, tobacco exposure or use of corticosteroids, antileukotrienes, antihistamines or immunotherapy. After a clinical evaluation and a skin prick testing, a nasal symptoms questionnaire was applied and a nasal washing was performed. Two hundred cells were examined (Giemsa and eosin yellowish) and apoptosis was identified by immunohistochemistry (Active-Caspase-3) (p20L18, sc-1225. Santa Cruz Biothecnology, Inc. Santa Cruz). RESULTS: The most common allergen was Dermatophagoides sp in 10 patients (91%). The total score of the nasal symptom questionnaire was 3 to 8 (median 6) in patients and 1 to 4 (median 2) in healthy subjects (p < 0.05, Mann Whitney U test). Frequency of eosinophils was 11 to 80% in patients and 0 to 1% in healthy subjects (p < 0.05, Z for proportions). Apoptosis was less frequent in patients (0 to 5, median 2) than in healthy subjects (4 to 16, median 8) (p < 0.01, Mann Whitney U test). CONCLUSIONS: Perennial allergic rhinitis may be associated with decreased apoptosis in the nasal mucosa. PMID- 16268183 TI - [Montelukast: new therapeutic option in patients with nasal polyps associated to respiratory allergic disease]. AB - BACKGROUND: The physiopathogenic mechanism of nasal polyposis is unknown. Chemical mediators found in nasal polyposis are: histamine, serotonin, leukotrienes (LTC4, LTD4, LTE4, LTB4), norepinephrine and possibly prostaglandin D2. Leukotrienes that mediate bronchoconstriction as well as chemical mediators of inflammation are observed in the nasal disease associated to polyposis. HYPOTHESIS: Antileukotrienes might play a significant role in controlling polyposis and symptoms due to sinonasal disease. They represent an alternative to conventional treatments with oral steroids as well as to operations in the long term control. OBJECTIVE: This study is a one year investigation that evaluates the important role of leukotriene receptor antagonists on nasal polyposis, with or without association to any allergic disease. Likewise, it compares its use with the conventional therapy used in the above mentioned disease. PATIENTS AND METHODS: We included 30 patients, 12 men (40%) and 18 women (60%), with ages between 16-45 years old. The mean age was of 20.7 years. Systematic assessments were made at the beginning of the study, as well as in the third, sixth, ninth, and twelfth month. Objective assessments, computed axial tomography of paranasal sinuses, allergological tests, nasal and bronchial symptoms evaluation, and interleukines, eosinophils, and immunoglobulin determinations were made. RESULTS: There was an improvement of nasal symptoms in the patients of group 1 (montelukast plus beclomethasone). They included the respiratory ones (p < 0.05), the nasal washing cells (p < 0.05), and the peak flow (p < 0.05) at the third, sixth, and twelfth months, respectively. There was no significant difference with the improvement obtained in the patients of group 3 (surgery-montelukast beclomethasone) (p < 0.05). There was no significant change in group 2 regarding the baseline in the studied variables (p > 0.05). PMID- 16268184 TI - [Increased accumulated incidence of allergic rhinitis in adolescents of Morelia, Michoacan, and its relation to satisfaction of vital and affective needs]. AB - OBJECTIVE: To determine the incidence (preliminary) of allergic rhinitis in a population-based sample of adolescents and its relation on how they meet basic and affective need satisfaction. MATERIAL AND METHODS: We used ISAAC survey to assess rhinitis-related symptoms in a population-based sample of adolescents from Morelia, Michoacan, Mexico. Some items were added to the survey to evaluate how they meet basic and affective needs. Random sample included 1,333 adolescents aged 11-16 years from several schools of Morelia, Michoacan. Allergic rhinitis impact on basic and affective need satisfaction was assessed. Relative risk (RR) and 95% confidence interval (CI) were estimated. RESULTS AND DISCUSSION: Until 2003, such disease was diagnosed in 12% of cases; however, the number went up after the survey: rhinitis-related symptoms were seen in 46% of those polled, of which prevalence, sensitivity and specificity rates were 27, 90, 70%, respectively. More symptoms were reported from July through December. Sixty percent of adolescents with allergic rhinitis reported sleep disturbances, with adverse effects on bed rest period (RR 2.64) [1.01-6.94]. Well-balanced diet was seen in 62% of adolescents with such disease (RR 1.56) [1.34-1.79]. Sixty-seven percent of such teenagers said that the disease does no interfere with their performance in school (RR 1.41) [0.61-3.24] and 22% felt unaccepted (RR 1.8) [0.795-16.63]. CONCLUSIONS: The estimation of incidence of allergic rhinitis found in this study is higher than in other reports. Basic and affective need satisfaction was affected; thus, implementation of psychotherapy within a multi disciplinary approach is required. PMID- 16268185 TI - [Psychological factors contributing to asthma in asthmatic children and adolescents and their parents]. AB - INTRODUCTION: Nowadays, psychosocial risk factors may contribute to asthma related problems among children, adolescents and family members; such disorders are described in this article. METHODS: A descriptive study, which included 50 children and their parents who took part in the educational program of Hospital Pediatrico Universitario Juan Manuel Marquez from 1997 to 1999, was performed. A survey, designed to establish psychological factors in children during both generally and symptom exacerbations, activities carried out with or without symptoms, and parental attitudes during illness episodes, was given to each one of the participants. Values were expressed in percentages. We used the Crosstabs to explore the relationships among variables. RESULTS: Anxiety, among other psychological stressors, prevailed in children (92%). Day-to-day activities were seriously affected in children with and without symptoms of illness: play 58%, exercise 60%. Children who received less parental support had severe persistent asthma, up to 20%. CONCLUSION: Symptoms severity is associated with child well being and negative attitudes of parents. PMID- 16268186 TI - [Safety of topical tacrolimus and pimecrolimus in children with atopic dermatitis]. AB - The use of topical immunomodulators in pediatric patients is an important topic in the clinical practice. Their prescription for chronic diseases suggests the necessity of evaluating their efficacy and safety profile in a long term period. In children they can develop systemic adverse events after their application, though sometimes they are useful to reduce the long consumption of other drugs, as topical steroids, or to have influence in the critical aspects of immunomodulation. Pimecrolimus and tacrolimus are two topical calcineurin inhibitors, from which there are several reports that support their efficacy in pediatric patients with atopic dermatitis. Recently, the FDA issued a recommendation for their topical use in a sporadic way in two years old children or older that have moderate to serious atopic dermatitis and that have not responded to other treatments. This article shows the results of several studies in which these drugs have been applied for a long time in children with atopic dermatitis. The more frequent adverse effects were: infections, pyrexia, burning, pruritus, erythema, and papules in the application area. In suckling babies they were: dry skin, pruritus, infections, constipation, erythema, and papules. Even when these adverse effects have been reported with relative frequency, their controlled use in concrete clinical conditions is still a therapeutic option and they should be considered particularly useful in the treatment of atopic dermatitis without positive reaction to other treatments in children older than two years, during short periods and in cases in which immunocompromised situations have been ruled out. PMID- 16268187 TI - Forgetting something. PMID- 16268188 TI - Naso-respiratory impairment and development of dento-skeletal sequelae: a comprehensive review. PMID- 16268189 TI - Cranial strains and malocclusion: II. Hyperextension and superior vertical strain. PMID- 16268190 TI - Practice philosophy: orthodontist/dento facial orthopedist part IV. PMID- 16268191 TI - Distalization in fixed mechanics. PMID- 16268193 TI - Weigh the digital "WOW" factor. PMID- 16268192 TI - Cleaning orthodontic bands after cementation. PMID- 16268194 TI - Above and beyond our call of duty. PMID- 16268196 TI - Growth hormone treatments for adult disorders. PMID- 16268195 TI - Growth hormone therapy effective in treatment of variety of hormone deficiency disorders in children. PMID- 16268197 TI - Important considerations in human growth hormone therapy delivery. PMID- 16268198 TI - Economic model shows savings through finer dosing increments. PMID- 16268199 TI - Managed care executives see utility in economic modeling for specialty injectables. PMID- 16268200 TI - Cost effectiveness and cost-savings opportunities associated with human growth hormone. PMID- 16268201 TI - A problem-solving strategy for teaching biosciences to nursing majors. AB - The author outlines a problem-solving strategy for teaching human anatomy, physiology, and microbiology to nursing majors. In Spring 2005, students improved their academic performance in comparison to Spring 2003. The author plans to further expand and analyze this teaching approach during the next two semesters. PMID- 16268203 TI - Student success strategies. PMID- 16268202 TI - Innovative instructional strategy using cinema films in an undergraduate nursing course. AB - Educators can develop innovative instructional strategies to engage students within the philosophical framework of Constructivism. To that end, the authors used films--Hollywood movies--to enhance their curriculum on neurological and psychopathological illnesses. During the fourth quarter of a seven-quarter associate degree nursing program, students developed case studies of the disorders portrayed in selected films. The authors outline the methods used to implement this approach and discuss evaluations from student and faculty perspectives. PMID- 16268205 TI - Every silver lining has a cloud. PMID- 16268204 TI - The impact of stress on academic success in college students. AB - The purpose of this article is to provide a discussion on stress and how it can prevent students from being successful in fulfillment of their educational goals. The literature is supportive of the fact that stress places demands on an individual, and in response to the stress, the body attempts to adapt to the stressful experience to maintain a sense of normalcy (Selye, 1974). Another common theme in the literature is that college students are faced with a unique set of stressors that may be overwhelming, thus altering the ability to cope with a situation. Strategies to reduce stress have been associated with academic success in college students (Dziegielewski et al., 2004). PMID- 16268206 TI - A balance between good and evil. PMID- 16268207 TI - Rhode Island Office of the State Medical Examiner: pathology in the public interest. PMID- 16268208 TI - Adult forensic psychiatry in Rhode Island. PMID- 16268209 TI - Forensic odontology and the postmortem identification process. PMID- 16268210 TI - Child and adolescent forensic psychiatry in Rhode Island. PMID- 16268211 TI - Forensic pediatrics. PMID- 16268212 TI - Rhode Island Child Death Review key findings: 2000-2002 deaths. PMID- 16268213 TI - Rhode Island's Child Death Review Team. PMID- 16268214 TI - The GENESIS project: culture change at the Miriam Hospital. PMID- 16268215 TI - [Could the development of post-traumatic disorder can be prevented by drugs?]. PMID- 16268216 TI - [Is manipulation treatment effective?]. PMID- 16268217 TI - [Prevention of allergies in the population]. PMID- 16268219 TI - [Prosa and medicine]. PMID- 16268218 TI - [Nephropathia epidemica]. PMID- 16268220 TI - [Resuscitation! Academic Medicine is in trouble]. PMID- 16268221 TI - [Vitamin D--multifunctional hormone]. PMID- 16268222 TI - [Atypical Parkinsonian syndromes, a challenging group of diseases]. PMID- 16268223 TI - [What is functional instruction for a patient?]. PMID- 16268225 TI - [Treatment of locally spread prostate cancer]. PMID- 16268224 TI - [Re-expansion of spontaneously collapsed lung and pulmonary edema]. PMID- 16268226 TI - [Castration of prostate cancer patients and cognition]. PMID- 16268227 TI - [Care of alcohol addiction]. PMID- 16268228 TI - [Red eye and a graduation ceremony]. PMID- 16268229 TI - The meaning of incontinence in dementia care. AB - Exploring ways of helping dementia sufferers and their carer's cope at home is central to government philosophy and legislation (Department of Health, 2001). The success of such support relies on having an accurate understanding of the nature and extent of the illness, and the caregiving experience. This article recognises and validates the pre-eminent role of spouse caregivers in the day-to day maintenance of dementia sufferers. In so doing, it promotes a caregiver coping paradigm as the most sensible and effective approach to formulating clinical assessment and service delivery. There is a dearth of research exploring the influence of incontinence on caregiver coping. This is surprising bearing in mind the seemingly obvious difficulties it creates for the caregiver. This relationship, described and discussed from a caregiver perspective, provides a valuable example of the need for a paradigm shift in the way professionals and services support and care for people with dementia and their spouses. PMID- 16268230 TI - Advocating for the rights of the mentally ill: a global issue. AB - The media has shared with the American public horrors of abuse towards prisoners of war in recent newscasts. Prisoners, and others in vulnerable positions, can fall victim to neglect and abuse at the hands of their captors. Often, human rights can be violated. Therefore, people in vulnerable positions need advocates who will defend and protect their rights. The role of advocate is familiar to nursing professionals. In the mid-1980's advocacy grew attention due to disclosure of human rights abuses in mental health facilities. Today, worldwide, nursing leaders continue to confront abuses in multiple settings. In the United States, the Human Rights Authority was specifically developed to address issues of abuse of persons with physical disabilities and or mental illness. Globally, nurses will want to support the development and enforcement of policies that protect persons in vulnerable positions. PMID- 16268231 TI - Clinical-academic partnerships research: converting the rhetoric into reality. AB - An increasing recognition of the importance of research-based practice to the nursing profession has led to a number of strategies designed to increase the utilization and conduct of nursing research. The transfer of nursing education from hospitals to universities occurred partly in response to the identified theory-practice gap. Subsequently, a significant investment in joint clinical academic positions and clinical professorial positions has been made with the intention of bridging the gap between the tertiary sector and the clinical field. Anecdotal evidence suggests that neither strategy has achieved the desired degree of success. The available literature suggests that nurses do not tend to become involved in the conduct of research, nor do they readily utilise research findings in their practice. It is hypothesized in this paper that this reflects the strong cultural differences between the clinical and academic worlds in nursing. The aim of this paper is to discuss the impact of these cultural differences and describe specific principals that could contribute to significant cultural change and the bridging of the academic-clinician divide. PMID- 16268232 TI - Southeast Asian refugee children: a school-based mental health intervention. AB - One particular focus of refugee studies in the United States has been the violence experience of Southeast Asian (S.E.A.) refugee children and its impact on mental health and school adaptation. Although virtually all researchers have found that the children have high rates of depression and/or post-traumatic stress disorder, findings concerning successful school adaptation have been inconclusive. Even so, concern has been generated on how to best meet the children's mental health needs. The purpose of our study was to provide an eight week school-based program that was designed to reduce depression symptoms of S.E.A. refugee children. Specifically, this collaborative program addressed refugee adaptation issues, children's culture and the development of coping skills. All of the children were screened for depression using the Children's Depression Inventory (CDI). Analysis of CDI data revealed that children's depression scores had a significant decrease between screening times 1 (approximately one month before the intervention) and 2 (fourth week of the intervention), 1 and 3 (eighth week of the intervention) and 1 and 4 (one month following the intervention). Globally, culturally sensitive mental health school based programs may be an appropriate intervention to assist immigrant and refugee children in making a successful adaptation to host countries. PMID- 16268233 TI - An anthropological hybrid: the pragmatic arrangement of universalism and culturalism in French mental health. AB - As in most European countries, the mental health of immigrants in France has recently been the subject of scientific scrutiny. Since the end of World War II voluntary special mental health services for migrants and refugees have been created in France and especially in Paris, but none has been based on epidemiological data. Generally, this lack of objective data gave rise to the assumption that many immigrants might not be getting the type of services they required. The birth of a new type of service (e.g. for migrants, refugees, ethnic groups, trauma and torture victims) was a political reaction to what was, at the time, expressed as an essential unmet need regarding this very specific population. In this article we review, from an anthropological point of view, the different paradigms that have prevailed over the last 50 years. PMID- 16268234 TI - Politics of healing and politics of culture: ethnopsychiatry, identities and migration. AB - Ethnopsychiatry is today a contested field, in which concepts and terms such as ethnicity, identity, culture, citizenship, traditional therapies or symbolic efficacy are used in a very controversial way. Recent accusations of'racism' against some ethnopsychiatrists have contributed to making more obscure the deep roots of these issues and controversies. Little attention has been paid to analysing the complex legacy of colonial psychiatry, as well as the relationships among current definitions of 'culture' and 'belonging', post-colonial subjectivities and migration. In this article, the authors briefly analyse the contributions of Italian ethnopsychiatry and investigate the hidden expressions of racism and prejudice still characterizing mental health workers' attitudes toward immigrants. It is argued that a 'generative' and community-based ethnopsychiatry can challenge the hegemony of western psychiatry and improve the quality of therapeutic strategies. PMID- 16268235 TI - Historical reflections on mental health care in Sweden: the welfare state and cultural diversity. AB - This article discusses historical reflections on the response of Swedish mental health care to cultural diversity and immigration and our views regarding future directions for clinical care, research and training. Sweden has become increasingly multicultural through immigration. Mental health care in Sweden faces the challenges of encountering cultural diversity and the mental health consequences of forced immigration, acculturation, and refugee trauma. In our view, Swedish mental health care is at a crossroads: either it takes up the challenge raised by immigration of an increasing cultural diversity or it satisfies itself with rhetoric, thus leaving reality at the margins. Equity regarding access to mental health care in Sweden today must include an acceptance of, and interest for, the diversity of the population. PMID- 16268236 TI - Multicultural mental health services: projects for minority ethnic communities in England. AB - Black and minority ethnic (BME) communities form 7.8% of the total population of the U.K. Many of these communities face a variety of disadvantages when they access, or are forced to access, statutory mental health services under the National Health Service. Efforts have been made to address these problems by developing projects both within statutory mental health services and in the non governmental ('voluntary') sector. This article describes some of these projects located in England, drawing out the themes and models that underlie their approaches, and discusses the lessons that can be learned from the U.K. experience. PMID- 16268237 TI - Mental health services in a multicultural society: interculturalization and its quality surveillance. AB - The purpose of this article is to present a model to promote and assess interculturalization of mental healthcare services in western multicultural society. We define interculturalization as the adaptation of mental health services to suit clients from different cultures. The suggested measures aim to introduce changes in four contexts: (i) the clinical interface or the relations between the immigrant patient and the healthcare workers and the treatment team; (ii) organizational adaptations required in the treatment context of the mental healthcare facility; (iii) the relation between the mental health facility and the ethnic communities; and (iv) the relation between the mental healthcare system, other facilities and society at large. To monitor the desired changes, the model describes qualitative and quantitative criteria and indicators to be applied in the four contexts. PMID- 16268238 TI - The Hong Fook experience: working with ethnocultural communities in Toronto 1982 2002. AB - The experience of the first 20 years of a community mental health service for the Asian communities in Toronto, the Hong Fook Mental Health Association, is presented within the context of the overall development of mental health services to ethnocultural minorities in Toronto and Canada. The various people involved and the relationships with other stakeholders are described. Through the discussion, the socio-political and cultural factors contributing to its successes and failures are highlighted. The future for Hong Fook, and the mental health system at large, is considered. The article finishes with an outline of the philosophical basis that has sustained Hong Fook's development. PMID- 16268239 TI - Overview of culturally-based mental health care in Vancouver. AB - This article is a description of how cross-cultural services in mental health have evolved in Vancouver. With 49% of Vancouver's total population described as a 'visible minority' by Statistics Canada, it has been essential for the city, in its efforts to provide health care that is accessible, available and acceptable to all, to develop health care that acknowledges racial and cultural diversities. Vancouver's Cross Cultural Mental Health Services had their beginnings over 25 years ago. The services encompass both formal and informal sectors of the healthcare system, are provided at primary, secondary and tertiary levels of healthcare delivery and are available through hospital- and community-based services. With recent regionalization of British Columbia's health services, the cross-cultural mental health service has experienced increased coordination under the administration of the Vancouver Coastal Health Authority (one of six British Columbia health regions). The initial elements of a cross-cultural mental health service consisted of the Vancouver Association for the Survivors of Torture, the Cross-Cultural Clinic at Vancouver General Hospital, and the Multicultural Liaison Workers Program of the Vancouver Community Mental Health Service. Collaboration and partnerships between the formal and informal sectors support each other, bridge gaps in services and provide a milieu for growth and development. PMID- 16268240 TI - A qualitative needs assessment for development of a cultural consultation service. AB - Cross-cultural mental health services were assessed using qualitative interviews and focus groups of 43 mental health clinicians and program directors in one of the most ethnically integrated cities in the U.S. The commonly used strategy of ethnic matching between clinician and patient was found to be difficult to effectively apply to an ethnically diverse and highly integrated patient population. Information from cultural-competency training was also found to be difficult to apply, particularly due to time limitations and language barriers. Implementation of a cultural consultation service, which uses in-depth cultural evaluations and case-based learning, may help bridge these service gaps. PMID- 16268241 TI - Are physical therapists fulfilling their responsibilities as diagnosticians? PMID- 16268242 TI - The relationship between forefoot, midfoot, and rearfoot static alignment in pain free individuals. AB - STUDY DESIGN: Correlational study. OBJECTIVES: To determine whether, and to what degree, a relationship exists between forefoot angle and weight-bearing midfoot and rearfoot position. BACKGROUND: There have been conflicting reports with regard to the degree to which the structure of the foot may influence the function. The influence of forefoot structure on weight-bearing midfoot and rearfoot position has not been extensively investigated. METHODS AND MEASURES: Fifty-one healthy subjects participated in this study (26 male and 25 female). Forefoot angle was measured in prone as varus (positive numbers), neutral (0), or valgus (negative numbers). Navicular drop was measured from subtalar joint neutral to unilateral standing relaxed. Rearfoot angle was measured in relaxed single-limb stance as the angle between a line that bisected the calcaneus and a line that bisected the lower third of the leg. The relationships between forefoot angle and navicular drop, and between forefoot angle and relaxed rearfoot angle, were investigated. The same relationships were also investigated in the neutral forefoot subgroup when the sample was divided in 3 subgroups based on 1 standard deviation of forefoot angle. RESULTS: There is a significant relationship between forefoot angle and relaxed rearfoot angle (r = 0.52, P < .001), as well as between forefoot angle and navicular drop (r = 0.55, P < .001), in the whole sample (n = 51). Average degrees of forefoot angle in the neutral subgroup (between 1.0 degree and 8 degrees of varus) are not associated with predictable positions of relaxed rearfoot angle (r = 0.19, P = .24) or navicular drop (r = 0.01, P = .96). CONCLUSIONS: Based on the results of this study, there is a significant relationship between forefoot angle and relaxed rearfoot angle, as well as between forefoot angle and navicular drop, in healthy subjects. These relationships were not found when forefoot varus values were within a standard deviation of the sample mean. PMID- 16268243 TI - Performance of the craniocervical flexion test in subjects with and without chronic neck pain. AB - STUDY DESIGN: Cross-sectional comparative study. OBJECTIVE: To compare the performance of the deep cervical flexor muscles on the craniocervical flexion test (CCFT) in individuals with and without neck pain. BACKGROUND: Significant weakness of the superficial neck muscles is often found in patients with neck pain. However, there is scant work on deep cervical flexors performance in subjects with chronic nonspecific neck pain. METHODS AND MEASURES: Twenty asymptomatic subjects and 20 subjects with chronic neck pain (duration, > 3 months) were recruited. The CCFT was performed with the subject supine and required performing a gentle head-nodding action of craniocervical flexion (indicating yes) for 5 incremental stages of increasing difficulty. Each stage was held for 10 seconds, as guided by the pressure biofeedback unit. The data used for analysis were the highest pressure level that each subject was able to hold for 10 seconds, up to a maximum of 30 mmHg. RESULTS: Reliability data obtained on 10 asymptomatic subjects indicated that the CCFT was reliable, with a kappa coefficient equal to 0.72. Subjects with chronic neck pain had significantly poorer (P < .001) performance on the CCFT (median pressure achieved, 24 mmHg) when compared with those in the asymptomatic group (median pressure achieved, 28 mmHg). CONCLUSIONS: The results of this study demonstrated that patients with chronic neck pain had a poorer ability to perform the CCFT when compared with asymptomatic subjects. The study adds to the evidence that poor ability to perform the CCFT may be clinical evidence of an impairment that characterizes neck pain, regardless of origin. PMID- 16268244 TI - Current status and correlates of physicians' referral diagnoses for physical therapy. AB - STUDY DESIGN: Randomized multicenter retrospective chart review of medical referral diagnoses and corresponding referral, patient, and physician demographic data. OBJECTIVE: To examine the information content of medical referral diagnoses provided to outpatient physical therapists with respect to physician and patient characteristics. BACKGROUND: Previous studies indicate that physicians commonly provide nonspecific referral diagnoses to physical therapists. The effects of patient and physician characteristics on information contained in referral diagnoses are not well elucidated. METHODS AND MEASURES: A team of blinded raters categorized the information content of referral diagnoses (n = 2183) using a classification system adapted from a previous study. RESULTS: One third (32%) of analyzed diagnoses were anatomically oriented and reported specific pathology. These specific diagnoses were provided significantly more commonly by specialist physicians (odds ratio [OR], 3.4; 95% confidence interval [CI], 2.7-4.2; P < .001), male physicians (OR, 2.2; 95% CI, 1.6-3.1; P < .001), both early- and late career physicians (P < .001), and for male patients (OR, 1.3; 95% CI, 1.1-1.6; P < .05). CONCLUSION: Nonspecific referral diagnoses are frequently provided to physical therapists by physicians. The practice of evidence-based physical therapy seems challenged by the high rate of nonspecific referral diagnoses. Physical therapists may also have the responsibility to conduct differential diagnosis of pathology more commonly than formally recognized by many state practice acts and third-party payers. PMID- 16268245 TI - The centralization phenomenon and fear-avoidance beliefs as prognostic factors for acute low back pain: a preliminary investigation involving patients classified for specific exercise. AB - STUDY DESIGN: Secondary analysis of a prospective cohort of patients with acute low back pain (LBP). OBJECTIVES: To determine if the centralization phenomenon and fear-avoidance beliefs predict measurement of pain and disability 6 months after entering the study. BACKGROUND: The centralization phenomenon and fear avoidance are predictive of future pain and disability. However, previous prognostic studies have not routinely included both measures in homogenous subgroups of patients with acute LBP. METHODS AND MEASURES: Patients completed self-report questionnaires and were evaluated and treated with treatment-based classification guidelines. Only the patients classified for specific exercise were included in this analysis (n = 28). Measures of disability and pain intensity were reassessed at 6 months by mail. Separate hierarchical regression models predicted measures of disability and pain intensity with the centralization phenomenon, fear-avoidance beliefs, and prespecified covariates. RESULTS: There were no significant differences in duration of symptoms, fear avoidance beliefs, and history of LBP based on the centralization phenomenon (P > .05). Patients reporting the centralization phenomenon were significantly more likely to have leg pain (P < .01). A regression model including initial disability, the centralization phenomenon, and fear-avoidance beliefs about work significantly predicted 6-month disability, explaining 49% of the total variance (P < .001). A regression model that included initial pain intensity and the centralization phenomenon significantly predicted 6-month pain intensity, explaining 29% of the total variance (P < .016). These factors also appeared to be clinically meaningful predictors of outcome, but lacked precision for immediate use in clinical settings. The following covariates were not included in the final regression models: presence of leg pain, history of LBP, and duration of LBP. CONCLUSIONS: Baseline elevation in fear-avoidance beliefs about work and lack of centralization phenomenon predicted higher disability. Baseline lack of centralization phenomenon predicted higher pain intensity. These results can only be generalized to patients with acute LBP classified for specific exercise. It will be necessary to independently validate these prediction models before they can be implemented in clinical settings. PMID- 16268246 TI - Criterion-related validity of a clinical measure of dorsal first ray mobility. AB - STUDY DESIGN: Test-retest methodological design using a sample of convenience. OBJECTIVE: To determine the criterion-related validity and the reliability of measuring first ray mobility with a ruler. BACKGROUND: Studies have questioned the accuracy of assessing first ray mobility by manual examination. Use of a ruler and adherence to strict guidelines in positioning of the patient may improve the measure. This study investigates the validity, and the intrarater and interrater reliability of measuring dorsal first ray mobility with a ruler while following recent recommendations to standardize the position of measurement. A valid and reliable mechanical device designed to measure first ray mobility was used as the validation criterion of measurement. METHODS: Three clinicians performed ruler measurement of dorsal mobility on 14 subjects. A separate examiner measured dorsal mobility with the mechanical device. Intraclass correlation coefficients (ICCs) and standard error of measurements (SEMs) were computed to quantify the intrarater reliability of both testing procedures and the interrater reliability of the ruler measurement. ICCs of agreement were also computed to determine the concurrent validity of the ruler measurement for each clinician. RESULTS: Mechanical device intrarater reliability ICC was 0.98 (SEM = 0.15 mm). Ruler intrarater ICCs were equal or less than -0.06 (SEMs = 1.1 mm); ruler interrater ICC was 0.05 (SEM = 1.2 mm). The ICCs of agreement between the mechanical device and ruler method ranged from -0.44 to 0.06. CONCLUSION: The ruler method of testing demonstrates poor reliability and validity as a clinical measure. PMID- 16268247 TI - Knee function and pain related to psychological variables in patients with long term patellofemoral pain syndrome. AB - STUDY DESIGN: Nonexperimental, descriptive study, including 2 independent samples. OBJECTIVES: To assess the levels of mental distress and self-perceived health in subjects with long-term patellofemoral pain syndrome (PFPS) compared to a group of healthy subjects, and the relationship between knee function and knee pain to these psychological variables. BACKGROUND: Psychological variables and those describing self-perceived health status have been given little focus in PFPS research. METHODS AND MEASURES: One group of 25 men and women between 19 and 44 years of age with unilateral long lasting PFPS, and a control group (n = 23) of healthy subjects (age range, 18-44 years) participated in the study. Knee function was assessed with the use of the Cincinnati Knee Rating System (CKRS) and the triple jump test, and knee pain was measured by a visual analogue scale (VAS). Self-perceived health and mental distress were assessed with the Coop Wonca Chart and the Hopkins Symptoms Checklist-25 (HSCL-25) questionnaire. RESULTS: The mean (+/- SD) score on the Coop-Wonca Chart was 2.02 +/- 0.73 in the PFPS group, compared to 1.20 +/- 0.53 in the controls (P < .001). HSCL-25 mean (+/- SD) scores were 1.46 +/- 0.47 and 1.08 +/- 0.18 (P < .001) for the PFPS and the control group, respectively. When analyzed with correlation statistics, CKRS and VAS scores were found to correlate to those of the Coop-Wonca Chart and HSCL 25 scores. CONCLUSION: Levels of mental distress were higher in the group with PFPS than in the control group, while levels of self-perceived health were lower. Our data indicate that the levels of knee pain and knee function correlate closely to the degree of mental distress and self-perceived health in individuals with PFPS. PMID- 16268248 TI - Proliferation and differentiation of human embryonic germ cell derivatives in bioactive polymeric fibrous scaffold. AB - Human embryonic germ cell derivatives, a heterogeneous population of uncommitted embryoid body derived (EBD) cells, were studied in a bioactive three-dimensional (3D) fibrous culture. Their proliferation, morphology, gene expression and differentiation were investigated to gain insights on development of 3D bioactive scaffold for pluripotent stem cells. The expansion of the EBD cells in 3D environment was significantly higher than their two-dimensional controls after 21 days. No apparent differentiation of the EBD cells cultured in the 3D environment, as indicated by histology and gene expression profile analysis, was evident. Extracellular matrix production was weak in the long-term 3D culture, and the EBD cells maintained their multilineage gene expressions for the period studied. When nerve growth factor (NGF) was surface-immobilized on the fibrous scaffold via chemically-modified Pluronic, the EBD cells cultured in this scaffold showed evidence of entering the neural pathway. An upregulation of tyrosine hydroxylase mRNA expression was observed when EBD cells were cultured in the NGF-immobilized fibrous scaffold, as demonstrated by real-time PCR and immunofluorescence staining. The study suggests the value of such fibrous 3D culture in manipulating stem cell proliferation/differentiation and as a model for developing a bioactive scaffold. PMID- 16268249 TI - Dendrimer/methyl methacrylate co-polymers: residual methyl methacrylate and degree of conversion. AB - Dendrimer/methyl methacrylate co-polymers were studied for use in dental composites. The aim was to determine the effects of methyl methacrylate concentration in the resin mixture and polymerization method on the degree of conversion and residual monomer content of the copolymers. Two dendrimers were studied, D12 with 12 reactive methacrylate groups and D24 with 24 reactive groups. The concentration of methyl methacrylate varied from 20 wt% to 50 wt% of monomers. Camphorquinone (CQ) was used as the light-activation initiator and 2 (N,N-dimethylamino)ethyl methacrylate (DMAEMA) as the activator, both in the quantity of 3.0 wt%. Three polymerization methods were used: photo polymerization, photo-polymerized immediately followed by post-polymerization at 120 degrees C for 15 min, and photo-polymerization followed by postpolymerization after 7 days. The degree of conversion was determined using FT-IR. Residual monomers were extracted with tetrahydrofuran and methanol and analyzed with HPLC. The highest degrees of conversion were 65 and 62%, and the lowest residual monomer contents 1.0 and 1.5% for D12 and D24, respectively. These were measured after heat-induced post-polymerization. For D12, increasing the proportion of methyl methacrylate decreased the degree of conversion and increased the residual monomer content after photo-polymerization. Post-polymerization enhanced the polymerization of the dendrimer co-polymers in respect of degree of conversion and residual monomer content. The present study suggested that the tested dendrimer/methyl methacrylate copolymers require heat-induced polymerization to reach the generally accepted levels of degree of conversion and residual monomers. PMID- 16268250 TI - Development of amine-containing polymeric particles. AB - The objective of this study was to synthesize and characterize particles as a drug-delivery platform for gliomas, a highly advanced and invasive stage of brain tumor with poor prognosis. Poly(aminoethyl methacrylate-co-methyl methacrylate) particles were prepared by suspension polymerization and poly(aminoethyl methacrylate-co-poly(ethylene glycol) methacrylate) particles were prepared by emulsion (w/o) polymerization. Amine groups of the particles were complexed with tetrachloroplatinate to form a cisplatin-like molecule. Particles were characterized with respect to size, zeta-potential, amine content, loading efficiency and drug release. Poly(aminoethyl methacrylate-co-methyl methacrylate) particles had diameters of below 10 microm, whereas the poly(aminoethyl methacrylate-co-poly(ethylene glycol) methacrylate) particles had diameters of approx. 1 microm. Poly(aminoethyl methacrylate-co-poly(ethylene glycol) methacrylate) particles had a more positive zeta-potential as compared to poly(aminoethyl methacrylate-co-methyl methacrylate) particles, although the amino-group content of both particles was almost equivalent. The net positive charge on the particles decreased after complexation with tetrachloroplatinate for both types of particles. Both particles had very high platinum-loading efficiency (>85%) and showed slow release of platinum over time. Particles had relatively low cytotoxicity (LC50 > 100 microg/ml) and demonstrated a high degree of association with cells. Complexation with poly(aminoethyl methacrylate-co methyl methacrylate) particles significantly reduced the toxicity of platinum. The poly(aminoethyl methacrylate-co-poly(ethylene glycol) methacrylate) particles have potential for being an effective drug-delivery platform and continued investigation is warranted. PMID- 16268251 TI - Morphology and metabolism of hepatocytes microencapsulated with acrylic terpolymer-alginate using gelatin and poly(vinyl alcohol) as extracellular matrices. AB - Microcapsules with good mechanical stability were prepared using an appropriate mixture of alginate and acrylic terpolymer. It was found from the microscopic observation that the microcapsules had a porous structure with interconnected pores, with a size of 50-150 nm. The results of the permeability experiment of microcapsules using FITC-dextrans showed that the capsule had a molecular mass cut-off of 120 kDa. The hepatocytes encapsulated in both alginate and acrylic terpolymer with gelatin and PVA rapidly aggregated in the core. The aggregated cells showed high albumin synthesis and ammonia removal, suggesting good metabolic function. PMID- 16268252 TI - Mechanical properties of collagen gels derived from rats of different ages. AB - In a previous study, we found that different collagen gels produced using collagen fibrils extracted from 1-, 4- and 8-month-old rat tails essentially influenced the morphogenesis of epithelial cells. More importantly, the youngest collagen gel induces the highest level of cell apoptosis. The objective of this study was to investigate mechanical properties of various collagen gels correlated to the rat ages. A rheometer and dynamic mechanical analyzer were used to measure shear and compressive properties of hydrated collagen gels. Experimental results obtained from both testing modes showed that older age related collagen gels possessed a larger elastic modulus, possibly due to the enhanced cross-linking degree. The moduli obtained in shear mode were 1.4-2.7 times greater than those in compression. The results of shear test and compressive test consistently indicated the age of rats did have a statistically significant effect on mechanical properties of hydrated collagen gels. PMID- 16268253 TI - A thin carboxymethyl cellulose culture substrate for the cellulase-induced harvesting of an endothelial cell sheet. AB - Engineered tissues constructed with two-dimensionally organized cells provide promising parts for reconstructing damaged tissues. Here we propose a new method for fabricating a 2D sheet made of an endothelial cell monolayer. First a culture substrate was prepared by treating the glass surface with an amine-terminated organosilicon derivative, followed by the covalent attachment of a thin carboxymethyl cellulose (CMC) layer. Fibronectin was immobilized onto the CMC coated surface to promote cell adhesion. These surfaces were characterized step by step by means of contact angle measurement and X-ray photoelectron spectroscopy. Porcine aortic endothelial cells adhered to the culture substrate and consequently formed a confluent monolayer. When the substrate-cell composite was immersed in a cellulase solution, a cell sheet was spontaneously detached from the substrate due to enzymatic digestion of the CMC layer. The cell-cell connections were well preserved in the cell sheet, even after detachment from the substrate, most likely due to the fact that cellulase is harmless to mammalian cells. The cell sheet could be transferred to other culture dish with the aid of a hydrophilic membrane support, retaining the proliferation activity of the cells. The results obtained in this study demonstrate that cellulase treatment of the CMC layer is a rational and efficient method for obtaining a 2D cell sheet. PMID- 16268254 TI - Effects of sub-toxic concentrations of camphorquinone on cell lipid metabolism. AB - The biological effects of camphorquinone (CQ), an initiator for light-polymerized resins, have been reported to relate to its ability to generate free radicals and cause radical-induced membrane damage via lipid peroxidation. However, the effects of CQ on lipids other than peroxidation may result in unfavorable tissue responses especially at concentrations that are not overtly toxic to cells. The purpose of the current study was to examine the effects of CQ on cell lipid metabolism at subtoxic concentrations, with or without visible light irradiation. HCP and THP-1 cells were exposed to CQ with or without light irradiation under clinically relevant conditions and lipid metabolism was analyzed using 14C labeling and thin-layer chromatography. We found that CQ increased synthesis of neutral lipids, such as triglycerides, from 7 to nearly 15% of the total and diglycerides from 2% to about 3% of the total in HCP cells, while synthesis of phospholipids, such as sphingomyelin, was decreased by 1-1.5%. In THP-1 cells cholesterol synthesis increased more than 2-fold and cholesterol ester synthesis increased more than 5-fold. Light-activated CQ did not differ significantly in terms of its bioactivity compared to no-light conditions. We conclude that CQ significantly altered the metabolism of several important structural lipids in two cell types at sub-toxic concentrations that are clinically relevant. These changes in lipid metabolism may in turn affect membrane integrity and permeability and possibly lead to significant changes in cell responses. PMID- 16268255 TI - 2-methacryloyloxyethyl N-butylcarbamate: a new co-monomer for synthesis of polyurethane hydrogels with improved mechanical properties for biomedical applications. AB - Hydrogels with tunable hydrophilic and mechanical properties were synthesized by the free radical polymerization of 2-hydroxyethyl methacrylate (HEMA) and 2 methacryloyloxyethyl N-butylcarbamate. The resulting hydrogels were investigated for their equilibrium water content, sessile drop water contact angles, gel fraction, mechanical properties and protein adsorption. Results indicated that co polymer hydrogels have good hydrophilicity and that, with the incorporation of the 2-methacryloyloxyethyl N-butylcarbamate, mechanical properties could be improved significantly. without affecting other important properties. Lysozyme and albumin adsorption experiments demonstrated that, similar to most hydrogel materials, the co-polymer hydrogels adsorb more lysozyme than albumin and that the adsorption was dependent on hydrophilicity. The control poly(HEMA) hydrogels were found to adsorb more protein than the co-polymer hydrogels; this is thought to be primarily a consequence of protein absorption rather than protein adsorption. PMID- 16268257 TI - Perspectives on psoriasis. PMID- 16268256 TI - Oral sustained delivery of diclofenac sodium using calcium chondroitin sulfate matrix. AB - Chondroitin sulfate (CS) is a potential candidate for colon-specific drug carriers. However, the readily water-soluble nature limits its application as a solid-state drug-delivery vehicle. In this study, the CS formation of a polyelectrolyte complex (PEC) with Ca2+ (CS-Ca) was adapted to retain CS in a solid form for use in a drug-delivery system. Pre-treated CS with poly(ethylene glycol) diglycidyl ether (EX-810) followed by complexation with Ca2+ was also tested (CS-Ca-EX). Diclofenac sodium was used as a drug probe to evaluate the performance of the drug-release behavior of the complexes. The amount of diclofenac sodium released was higher in simulated intestinal fluid (SIF) than in simulated gastric fluid (SGF) due to the anionic groups on CS or the higher solubility of drug itself in PBS. The release profile of diclofenac sodium from CS-Ca-EX was most notably sustained when compared to other groups. Enzymatic degradation by chondroitinase ABC of CS, CS-Ca and CS-Ca-EX exhibited a similar degradation mechanism and GPC revealed the dissolution rate of CS from the three matrix types was, in decreasing order: CS, CS-Ca, CS-Ca-EX. The synergy of the anti-inflammatory activity of diclofenac sodium in CS-based complexes was evaluated using the carrageenan-induced edema rat test. The percentage of swelling was lower for all experimental groups as compared to the control, untreated group. The anti-inflammatory activity of diclofenac in the CS matrix gradually increased up to 9 h but CS-Ca or CS-Ca-EX matrices showed less potency than the CS matrix in reducing inflammation. PMID- 16268258 TI - What is your diagnosis? Terry nails. PMID- 16268259 TI - Infantile psoriasis. AB - Psoriasis is a common inherited papulosquamous dermatosis that may be a diagnostic dilemma, particularly in infants and children. The treatment of children with psoriasis should be handled with caution and tailored according to the child's age, as well as to the extent, distribution, and type of psoriasis. PMID- 16268261 TI - Alcohol as a risk factor for plaque-type psoriasis. AB - The association between alcohol and the development of plaque-type psoriasis is complex and confusing because many of the initial studies did not control for confounding factors such as tobacco use. This article presents a literature review of the epidemiologic, case-controlled, and clinical studies that examined the relationship between alcohol and plaque-type psoriasis. Early studies showed no correlation between alcohol consumption and plaque-type psoriasis. However, as researchers began to control for confounding factors, study results often illustrated a significant correlation between alcohol use and psoriasis. Some studies suggested a relative risk factor of 8.01, particularly in men. However, the studies did not document an increased risk for plaque-type psoriasis in women who drank alcohol. We recommend that clinicians discourage patients with psoriasis from consuming alcohol, especially during periods of disease exacerbation. PMID- 16268260 TI - Tea tree oil. AB - Tea tree oil is a popular ingredient in many over-the-counter healthcare and cosmetic products. With the explosion of the natural and alternative medicine industry, more and more people are using products containing tea tree oil. This article reviews basic information about tea tree oil and contact allergy, including sources of tea tree oil, chemical composition, potential cross reactions, reported cases of allergic contact dermatitis, allergenic compounds in tea tree oil, practical patch testing information, and preventive measures. PMID- 16268262 TI - Partial remission of psoriasis following rituximab therapy for non-Hodgkin lymphoma. AB - Rituximab, a human-mouse, chimeric, monoclonal antibody that targets the B-cell CD20 antigen and causes rapid and specific B-cell depletion, is indicated for the treatment of low-grade or follicular, relapsed or refractory, CD20+ B-cell, non Hodgkin lymphoma (NHL). We report the case of a middle-aged woman with psoriasis who experienced a partial sustained remission of her psoriasis after treatment with rituximab for NHL and discuss potential pathophysiologic mechanisms for this unexpected effect in a condition known to be mediated by T cells. PMID- 16268263 TI - Subacute cutaneous lupus erythematosus associated with leflunomide. AB - A skin eruption consistent with subacute cutaneous lupus erythematosus (SCLE) occurred in a patient taking leflunomide for rheumatoid arthritis. The eruption resolved after discontinuation of the medication. Suppression of tumor necrosis factor (TNF)-effector mechanisms by leflunomide may have played a role in the pathogenesis of this disorder. PMID- 16268264 TI - Metastatic gastric adenocarcinoma presenting as an enlarging plaque on the scalp. AB - We present a case report of metastatic gastric adenocarcinoma in which the cutaneous metastases were the first sign of disease recurrence. A 73-year-old man presented with painless red plaques on his scalp and forehead. He was diagnosed with gastric carcinoma with metastases to perigastric lymph nodes 3 1/2 years earlier. Histopathology results revealed signet-ring cells consistent with gastric adenocarcinoma. The patient failed to respond to treatment with intralesional interleukin 2, previously reported to be effective, and expired 7 months later. PMID- 16268265 TI - Unilateral segmental Darier disease following Blaschko lines: a case report and review of the literature. AB - Darier disease (DD) is an autosomal-dominant skin disorder that is characterized by multiple keratotic papules, loss of epithelial adhesion, and abnormal keratinization. We describe an unusual case of late-onset unilateral segmental DD that follows the lines of Blaschko. Our patient did not exhibit other classic findings of DD. Our case and review of the literature suggest that lesions previously classified as acantholytic dyskeratotic epidermal nevi (ADEN) are actually unilateral segmental presentations of DD. PMID- 16268267 TI - The Woody Cohlmia Golf Tournament--a legacy in the making. PMID- 16268266 TI - Treatment of bullous pemphigoid with dapsone, methylprednisolone, and topical clobetasol propionate: a retrospective study of 62 cases. AB - Dapsone has been used successfully as adjuvant therapy for bullous pemphigoid (BP). The effectiveness of dapsone for this indication, however, remains controversial. We evaluated the effectiveness and adverse events of dapsone (1.0 1.5 mg/kg per day) in combination with oral methylprednisolone (tapering doses of 0.5 mg/kg per day) and topical clobetasol propionate (initially applied once daily on lesions only) in the treatment of BP. Sixty-two patients treated with this regimen were analyzed retrospectively. Patients were free of new blisters after a mean (+/- SD) of 22 +/- 13 days (median, 20 days). After 3 and 6 months of treatment, methylprednisolone could be reduced to less than 10 mg/d in 71% and 91% of patients, respectively; after 12 months of treatment, 53% of patients were in complete remission without receiving further therapy. Dapsone-related side effects were usually mild except in 3 patients (5%), 2 patients with anemia (hemoglobin level, <7 g/dL) and 1 with agranulocytosis. Our data suggest that dapsone used in combination with systemic and topical corticosteroids may be a relatively safe and effective treatment option for BP. PMID- 16268268 TI - Profile: Bill Nations, DDS, State Representative. Interview. PMID- 16268269 TI - Party in the park: ADA, CDC celebrate 60 years of community water. PMID- 16268270 TI - Community water fluoridation: 60 years of fighting tooth decay. PMID- 16268271 TI - HIPAA security rule FAQs. PMID- 16268272 TI - ADA question & answer: oral healthcare in Native American communities. PMID- 16268273 TI - Oral health and learning: when children's oral health suffers, so does their ability to learn. PMID- 16268274 TI - Case study of benzocaine-induced methemoglobinemia. PMID- 16268276 TI - [The centrality of the family as a resource for homecare: gender and generational perspectives]. AB - Given the critical situation of informal caregiving in Spain, we explore how in the current socio-economic and political context main caregivers value and understand their family members as a resource to support caregiving. This qualitative study had a postfeminist orientation and was developed in Mallorca (Spain) through individual interviews and focus groups with men and women home caregivers from three generations. The participants identified their families, understood as women, as their main source of help due to the emotional quality of care they provide. However, internal conflicts and gender inequities, among others, made us question the dominant discourse of family's privileged position to take care of dependent people. The implications of these results to public policy are discussed. PMID- 16268277 TI - [Inquiry on condoms in the basin of Rio Acre: carriage, package, use and the infection risk for STD]. AB - Descriptive-exploratory type study, carried with 168 inhabitants of the basin of Rio Acre - Brazil, which aimed to investigate preservative use and the risk for Sexually Transmited Diseases. For data collection it was used an application form with spontaneous participation and anonymate guaranty. As result, stands out the 64.3% masculine gender, 75.0% married/joined, 57.1% had incomplete first degree and 31.0% were illiterate. At that time 78.0% did not bring preservative when coming to the city, and 6.0% just came out seeking for sexual activity; 62.0% had already used the masculine preservative, with 67.2% of acceptance and 77.8% had kept it in the back pocket of the pants, with danger of damaging and infection risk and undesired pregnancy; 8.9% had had venereal disease, being 80.0% for the gonorrhea. Of these, 46.7% had negotiated respectively with pharmacist and doctor. Thus, 89.3% considered the prevention campaigns as great/good. PMID- 16268278 TI - [Nurses: collaborating agents in the execution of the institutional mission]. AB - The study aimed to investigate the strategy that can be used by health institutions, with respect to the nurse's optics, to motivate/value and to promote the nursing worker's training, while collaborating agent of the institution seeking for the execution of his/her mission. For the materialization of the same, we invested in a qualitative, descriptive and exploratory methodology, whose research instrument, semi-structured, was applied to 15 nurses, being guaranteed to them, the ethical and legal issues reasonable in that study. In concluding this study we verified that the main strategies that can be used in health institutions, under the nurse's optics, to motivate/ value and to promote the nursing worker's training, concerns to the dialogue need and the promotion of the professional growth. PMID- 16268279 TI - [Development of an expert system for identification of nursing diagnosis related to urinary elimination]. AB - The study involved an evaluation of the ALTURIN.EXP system and its improvement that culminated in the establishment and evaluation of a new system named ALTURIN.SDD. In the present work the evaluation phases of the ALTURIN.EXP and development of the ALTURIN.SDD systems are described. The ALTURIN.SDD system was developed by utilizing a "shell" program specially built for this project, the Program for Diagnose Determination (SDD, 1.0 Version), which had as a target Windows 95/98/Me platforms. The nursing diagnoses are those present in the NANDA 2001-2002 classification. This development experience shall be applied in the development of other specialized systems. PMID- 16268280 TI - [Applying situational leadership in emergency nursing]. AB - This study aimed to identify the correspondence of opinions between nurses and nursing staff members who work at the emergency unit regarding the leadership style performed by nurses and the style nurses must adopt considering the maturity level of nursing staff members and the care provided at the unit. Situational Leadership was adopted as a theoretical reference framework. In order to achieve these goals, instruments were elaborated and applied to 24 research participants. Results demonstrated that the leadership style nurses most frequently perform was E3 (participating), and that nurses should adopt leadership style E4 (delegating) in view of the nursing staff's level of maturity, suggesting that staff members present a high level of maturity (M4). PMID- 16268281 TI - [Analysis of the care delivered to hospitalized newborns according to the Winnicottian perspective]. AB - This work aimed to analyze the holding provided by nurse aides, nurses, mothers, and fathers to infants newborn hospitalized at the middle risk neonatal unit of a university hospital in the city of Sao Paulo. This is a qualitative descriptive study based on Winnicott's theoretical framework. Data were collected by means of videotaping. Results demonstrated that there are some professionals who fail in providing holding, whereas others provide it satisfactorily. Although parents provide adequate holding, at times they fail to do so. We believe that it is necessary that the caring practice must be reevaulated, so that all members of the team, and also mothers and fathers are able to provide adequate holding for the infant's development. PMID- 16268282 TI - [Caring for the elderly with dementia: a study of ambulatory nursing practice]. AB - The care provided by caregivers to elderly people with dementia is investigated having as objectives: to describe the characteristics of sixteen elderly and their caregivers in attendence at the neurogeriatrical ambulatory of Rio de Janeiro during 2004; to identify nursing and care diagnosis used by caregivers, through EDG, MEEM, AIVDs and AVDs applied in nursing assessments. The quantitative descriptive method was employed, as a study of each case, obtaining the results: in the client- Disorientation, aggressiveness, skin drought, urinary incontinence, disturbance in the family relationship; In the caregivers- inefficient caregiver/elderly communication, physical fatigue, lack of knowledge about care; accomplished types of care--feeding, medication and hygiene. In conclusion, educational nursing actions are necessary to improve the care quality to the elderly and their family. PMID- 16268284 TI - [The puerperal infection in a delivery center: occurrence and predisposing factors]. AB - The Delivery Center (DC) provides obstetric assistance centered on the parturient's needs according to scientific evidences. This study aimed to verify the occurrence of puerperal infection in the DC and to compare the clinical obstetric characteristics of the women readmitted in the hospital with puerperal infection to those who were not readmitted in the hospital. It is an exploratory and non-experimental research with retrospective data collection. The population was based on 51 puerparae who received assistance in the DC, from 2000 to 2003. The results demonstrated that among 10,559 deliveries, 0.16% of them presented puerperal infection and the obstetric assistance given in the DC was related to the puerperal infection, only when considering the duration of the labor. It was concluded that the obstetric assistance given by this model of care brings benefits to the women. PMID- 16268283 TI - [The experience of becoming parents of a premature infant: an ethnographic approach]. AB - The aim of this study was to understand the experience of the parents of premature newborn infant of very low birth weight, discharged from Neonatal and Pediatric Intensive Care of the University Hospital. It was conducted through qualitative approach, using ethnography as method, in its interpretative perspective. Data were collected using participant observation, and interviews with six parents. From this analysis emerged six categories and two cultural themes: "The capacity to be parents: fighting moments and growth" and "Caring and living together with a child". Parents lived all the process with fear and hope. They said this experience was very intense and transforming. They felt capable to take care of the child after hospital discharge, although recognized new obstacles in the social-cultural context to carry out the responsibility of being parents. PMID- 16268285 TI - [Prevalence of specific hypertensive disease of pregnancy in a public hospital of the city of Sao Paulo]. AB - This is a descriptive-exploratory study carried out from January to July 2002 with the purpose to know the prevalence of Specific Hypertension Disease in pregnancy (SHD) and its disorders. A total of 604 records of hospitalized women were retrospectively analyzed and 22 were diagnosed with SHD, that is, 3.64% of women presented this diagnosis. Of those, 45.45% were adolescents and 40.90% first pregnancy. In 86.36% the pathology occurred after the 20th week of gestation. Eclampsia, hypertension crisis, intrauterine fetal death, neonatal death, chronic fetal suffering and prematurity are one of the observed disorders during pregnancy. It was concluded that perinatal health assistance given to pregnant women in this region must be reviewed specially for the adolescents aiming at better perinatal health indexes. PMID- 16268286 TI - [Nursing service assessment: identification of process criteria in hospital accreditation programs]. AB - Efforts to ensure quality in healthcare, to elaborate patterns and criteria have been a challenge in Brazil. This study aimed to identify the process of evaluation criteria applied to the nursing services adopted by Accreditation programs, and classify them in administrative, assistential and teaching research attributions. The study was exploratory, descriptive and quantitative. Collected data from 7 companies accredited by the National Accrediting Organization leaded into 8 models, which were divided into A, B and C groups. Seventy-nine criteria were identified; 32 from assistential, 32 administrative and 15 teaching-research processes. The nursing service was evaluated with focus in the C group. A small amount of assistential and learning research in A and B groups, showing that an increase in criteria would be necessary in order to obtain more accurate measuring. PMID- 16268287 TI - [Heart failure: old syndrome, new concepts and nurse's role]. AB - Congestive Heart Failure (CHF) has been cause of increasing concern in the world, face its progressive prevalence and incidence and its social and economic repercussion, pointing to the need of revision of the syndrome concept and adoption of measures to reduce its economical and social costs. The objective of this study is to contextualize the comprehension of the physiopathology and the treatment of CHF and the nursing interventions to patients with this syndrome. PMID- 16268288 TI - [Nurse's image: review of the literature]. AB - The society's image about the nurses is permeated of stereotypes that include figures like saints, prostitutes, witches, heroines and are related to the function of assisting doctors and to a lack of social life. Moreover, the image that the nurses have of themselves and of their work is negative, causing frustration for the lack of autonomy found on the professional reality. Seeking for a better understanding of this problem, a literature review was proceeded, aiming to characterize and analyze the nurse's image. The national scientific production is considered small on the subject. Conclusions indicate that stereotypes negatively influence nursing practice and that in Nursing history it is found many reasons for the existing distortions. PMID- 16268289 TI - [Changes in nursing administration in supporting transplantation in Brazil]. AB - This historical and bibliographic study aimed to understand how Nursing was organized to support care in transplantation. The HISA, LILACS, BDENF, PERIENF and DEDALUS databases were consulted, and thirteen references were found, ten of which were scientific articles, two were master's dissertations and one was a doctoral thesis. The span of time chosen for study ranges from the date of the first kidney transplant in Brazil (1965), to the date of publication of the last scientific article found in the databases mentioned above (2003). After reading these articles, the ones that were similar in topic were grouped together, thus creating the thematic axis for the presentation of the results. The results showed that the Nursing profession has played an important and active role in transplants ever since the first procedure in 1965. PMID- 16268290 TI - [Nursing in life re-significance on the face of disease]. AB - The paper deals with the educational role played by the health care provider. The ideas exposed stem from a reflection made from a research on the resignification of life of individuals affected by diseases that entail changes to their lifestyle. The health care provider has a key role in this process, influencing the patient positively or negatively. The reflection approaches the subjective process of life resignification in the health/disease situation and the educational role of the nursing professional in performing their tasks. Teaching and learning influence the configuration of the individual's imagery, constituting thereby an important ingredient to the educational know-how in the professional practice. PMID- 16268291 TI - [Punctuating ideas on the methodological development of social representations in Brazilian researches]. AB - This text proposes an alternative approach to the methodological discussions on the studies of social representation in the field of nursing, which are usually anchored in an opposition between quantitative and qualitative methods. It is argued that such an opposition is just an apparent one, since it is possible to observe similar results in studies conducted with different methods. With a basis on the thought of several authors in the field of social representations, the question is treated from the standpoint of the distinction between a processual and a structural approach to the phenomenon of social representation. It is proposed a taxonomy that, in departing from those perspectives, takes into consideration the data collection and data analysis implied techniques, as well as the plan of the representation to which the researcher wants to get access. PMID- 16268292 TI - [Competence approach in teaching-learning process]. AB - The work relates the experience of the teaching-learning process based on the competence approach. The detail of that teaching process reveals the authors concern with the few occurrence of studies about the practical development of the competences theme in the Educational Practice and, point out to the need of more studies production in that dimension. PMID- 16268293 TI - [Patient safety and the paradox in medication use]. AB - Administering medication to patients is a complex process with various phases which contemplate a series of inter-related decisions and actions involving professionals from a number of disciplines as well as patients themselves. Errors may occur in any phase of such process. This article describes the indexes, terminology and classification of medication errors and determines the approaches which explain their occurrence, namely the person-centered approach and the system-centered approach. The aim of the paper was to contribute, through theoretical elements, to the strategic discussions concerning the improvement of care standards in health care institutions in our country, focusing on patient safety related to drug therapy. PMID- 16268294 TI - [Information system in nursing: interacion of tacit-explicit knowledge]. AB - The present article aims to trace some theoretical and conceptual considerations on information systems in nursing, seeking to point out the knowledge based on the clinical practice evidences to construct a model of system integrated to the conceptual structures, formed by the combination of three sciences: information, computing and nursing. This knowledge can systematically describe and explain the necessary phenomena to develop a comprehensive information system that contribute for nursing records improvement and to consolidate a mechanism to provide basic measuring of costs, quality, patient access to care, and results of this care. PMID- 16268295 TI - [Nursing representative entities in the State of Goias: a historical report]. AB - The Nursing representative entities in Goias have been poorly explored. This work aims to reconstitute the path of these institutions, in the context, in which they were created and their main remarks. It is a qualitative study, of historical - social type. The ABEn was founded in 1948, working in the "Sao Vicente de Paulo" school dependences. The COREn was created in 1975 and the SIEG in 1982. The COREn, as a professional legislative institution, agglutinates the ones who exercise the profession, adopting government determinations. The ABEn, since 1986, started to assume a more committed professional behave with subjects of the category and the population. The SIEG has been having small representativeness, because the nurses, usually, are linked in the other unions. PMID- 16268296 TI - [Poliomyelitis surveillance - Acute flaccid paralyses]. PMID- 16268298 TI - Moving forward as one family of medicine: MSMS priorities and initiatives. PMID- 16268297 TI - Recent affidavit of merit decision would have been trouble for physicians and plaintiffs. PMID- 16268299 TI - Complementary and alternative medicine attracting attention. PMID- 16268300 TI - The priceless benefits of membership. PMID- 16268301 TI - American physicians assurance: providing the resources and protection you need. PMID- 16268302 TI - CDC issues new influenza recommendations for 2005-2006 flu season. PMID- 16268303 TI - One family = one voice. PMID- 16268304 TI - Put care back into homes. PMID- 16268305 TI - Will the government's world-class aims give nursing research the boost it needs? PMID- 16268306 TI - Lack of breaks leaves nurses burnt out. PMID- 16268307 TI - Campus care. AB - As the academic year begins, nurses are set to help students living away from home for the first time with their health and emotional needs. Each educational institution organises student health services. Loneliness can be a particular problem for overseas students. Students can be at risk from diseases such as mumps and meningitis. Drug and alcohol misuse are serious concerns. PMID- 16268308 TI - Words of wisdom. AB - Most students experience good and bad mentoring throughout their training. Bad mentors can cause serious problems further down the line, particularly if they 'fail to fail' incompetent students. However, the pressure on mentors is immense and students may not have realistic expectations. The Nursing and Midwifery Council and the RCN are introducing measures to improve the overall experience of mentoring. PMID- 16268309 TI - Not-so-private practices. PMID- 16268310 TI - Handling with care. PMID- 16268312 TI - With an accent on prejudice. PMID- 16268311 TI - Gene politics. PMID- 16268313 TI - Save primary care. PMID- 16268314 TI - Working in care homes: a survey. AB - AIM: To explore the impact of policy changes on care homes and the provision of nursing in care homes. METHOD: Eight hundred Royal College of Nursing (RCN) members working in nursing care homes were surveyed by postal questionnaire in 2004. RESULTS: A 37 per cent response rate (n = 296) was achieved. Of these, 274 worked in homes registered to provide care for older people and/or those with dementia. Although the majority of respondents were happy in their posts, they said that they were not always able to meet residents' needs. They voiced concerns about inappropriate admissions, the assessment process and the need to fill beds to maintain income. Although 65 per cent of residents were state funded, almost 75 per cent of the homes charged these residents a top-up fee. CONCLUSION: Care homes should be given enough resources to ensure that the needs of residents are met. The RCN is developing a programme to lobby government for adequate resources for care home placements. PMID- 16268315 TI - Recognising the limitations of your genetics expertise. AB - The sixth article in this series aims to provide you with sufficient knowledge to refer a client for further genetic assessment. The focus is on the skills needed to recognise the limitations of one's own genetics competence, as described in the competency standard statement; this is that, at the point of registration, all nurses, midwives and health visitors should be able to recognise the limitations of their own genetics experience based on an understanding of their professional role in the referral, provision or follow up to genetics services. PMID- 16268317 TI - Administration of medicines. PMID- 16268318 TI - Negotiate in style. PMID- 16268316 TI - Emergency first aid for nurses. AB - This article provides guidance for nurses on giving first aid in the domestic, pre-hospital and clinical settings. It focuses on resuscitation, cardiac conditions, shock and haemorrhage. PMID- 16268319 TI - Tips for success. PMID- 16268320 TI - Frustrations in clinical practice. PMID- 16268321 TI - Clinical trials in sepsis. PMID- 16268322 TI - Acute renal failure on the intensive care unit. PMID- 16268323 TI - Recent advances in mechanical ventilation. PMID- 16268324 TI - Weaning from mechanical ventilation. PMID- 16268325 TI - Critical care outwith the intensive care unit. PMID- 16268326 TI - A review of the coroner system in England and Wales: a commentary. AB - The certification of deaths and their investigation is flawed and has not been subject to comprehensive revision for many decades; the current system is fragmented. Despite its historical 'stability', it is poorly understood by many who have to use it and the lack of supervisory structures within the system means that there is no leadership, accountability or quality assurance. No formal linkage to or communication with other public health services and systems exists, minimising its epidemiological value. There is a lack of clear participation rights in these processes for bereaved families. The standards for the treatment and support of the bereaved are woefully inadequate and have contributed in a major way to certain causes celebres. A report in 2003 suggested that death investigation should be a service that is consistent and professional, able to deal effectively with legal and health issues, work across the full range of concerns about public health and public safety and support, and audit the death certification process. The role of those supporting the current system must be properly established in a framework of accountability. PMID- 16268327 TI - Supporting revalidation: methods and evidence. AB - The aim of revalidation is to reassure the public that doctors are up to date and fit to practise. The Royal College of Physicians has developed various programmes of work, both ongoing and under development, in support of revalidation. The purpose of this work is to provide guidance for doctors about the standards of practice expected and the types of evidence required for revalidation. A range of methods and tools are outlined that could be used by doctors to demonstrate their commitment to evaluating and reflecting on the way they practise medicine. These methods include ways of assessing both professional standards and clinical competence. Although the conclusions of the Chief Medical Officer in terms of the absolute requirements for revalidation are still unknown, the College is committed to supporting its Members and Fellows to prepare for revalidation through setting educational, professional and clinical standards in medicine. PMID- 16268328 TI - Risk communication in the clinical consultation. AB - Modern healthcare and modern societies are facing up to the need for greater engagement of patients in treatment decisions. Shared and informed decision making is replacing traditional paternalistic approaches to decisions; health policy both reflects and drives these changes. A critical contribution to better informed decisions by patients is the effective communication of risk in the clinical consultation. This is not straightforward, but there is a growing evidence base to improve performance in this area to the benefit of both patients and clinicians. The purpose of this review is to provide an accessible and practical guide to better communication of risk by clinicians. PMID- 16268329 TI - Pharmacopolitics and deliberative democracy. AB - Setting priorities in healthcare has become a highly politicised activity. Traditionally it has been undertaken by government and the health professions but there is an increasing imperative to involve the public. The National Institute for Health and Clinical Excellence (NICE) has established a Citizens' Council as an attempt to capture the informed views of the public in shaping the Institute's social value judgements. Although, in the future, better ways of involving the public may emerge, NICE's approach represents one way in which ordinary citizens can engage in the process of prioritising healthcare. PMID- 16268330 TI - Molecular genetics goes to the diabetes clinic. AB - Diabetes has historically been thought of as a medical specialty which primarily deals with treatment rather than diagnosis. Molecular genetic testing can now be used to make a diagnosis of the 1-2% of all diabetic patients with monogenic diabetes. Making a diagnosis of monogenic diabetes is important as it can have a dramatic effect on the treatment a patient should receive: glucokinase MODY patients need no treatment; HNF1alpha MODY patients are very sensitive to low dose sulphonylureas; and patients with neonatal diabetes due to Kir6.2 mutations, despite being insulin dependent, can discontinue insulin and be well controlled on high dose sulphonylurea tablets. The challenge for diabetologists is to use clinical skills to detect these monogenic patients whose care will be greatly helped by the treatment changes that follow molecular genetic testing. PMID- 16268331 TI - Don't look now! Pain and attention. AB - Attention and pain are linked inexorably. The manipulation of attention, via either distraction or focused attention, has been used as a therapeutic initiative for generations. Imaging evidence and clinical observations demonstrate that attention can be altered with associated changes at the cortical level and this may have positive or negative effects on the individual. New theories suggest that cortical remapping and visual attention may play key roles in a cortical model of pain specifically involving the motor control system. Within this system, the relationship between allocentric (external) and egocentric (internal) stimuli are managed; where conflict occurs, somaesthetic disturbances may be generated. If an individual pays too much attention to such sensory disturbances, then they may report the disturbances as abnormal symptoms, which may explain the diverse symptomatology of fibromyalgia. The use of a therapeutic optokinetic device to correct existing imbalances in the motor control system is also discussed. PMID- 16268332 TI - HIV/AIDS in the transitional countries of Eastern Europe and Central Asia. AB - In the 1990s, HIV/AIDS became a major threat to health, economic stability and human development in countries in eastern Europe and central Asia. Social, political and economic transition exacerbated the structural conditions that allowed HIV/AIDS to flourish as dramatic changes led to increasing drug injection, economic decline and failing health and healthcare systems. There is a need to address the professional and ideological opposition - even in countries considered to be fully functioning democracies - to evidence-based public health interventions like harm reduction, coupled with treating HIV/AIDS for all those in need, if countries are to provide a more effective response. PMID- 16268333 TI - Genetic predisposition to cancer. AB - Over recent decades a number of genes causing predisposition to cancer have been identified. Some of these cause rare autosomal dominant monogenic cancer predisposition syndromes. In the majority of families, the increased incidence of cancers is due to a multifactorial aetiology with a number of lower penetrance cancer predisposition genes interacting with environmental factors. Identification of those at increased risk of cancer on account of their family history is important, as genetic testing, enhanced surveillance, prophylactic surgery and chemoprophylaxis may be indicated. In this article the issues surrounding genetic predisposition to cancer are considered by examining two common cancers: colorectal and breast cancer. PMID- 16268334 TI - Patients and doctors: rights and responsibilities in the NHS. AB - Whilst patients and doctors each bring a collection of rights and responsibilities with them into the consulting room, these parties must not be seen in isolation. The government of the day and its representatives have an absolute right to influence the future direction of the health service but they also have very grave responsibilities. PMID- 16268335 TI - Patients and doctors: rights and responsibilities in the NHS. AB - Our society is very concerned with endowing and protecting people's rights and the existence of a National Health Service in this country has allowed such concepts to be applied to medical care. It is not clear, however, whether the concept of rights helps either doctors or patients, and it may in fact be misleading or even damaging. A discussion of the suitability of rights ethics to the healthcare context is presented, particularly highlighting the problems of attributing positive rights. The way in which rights create corresponding duties and responsibilities, and the particular problems that this creates for our health service is also addressed. Finally, in the light of the ongoing changes in the doctor-patient relationship, the future balance of rights and responsibilities in healthcare and the impact this will have on clinical decision making is discussed. PMID- 16268336 TI - Cardiac amyloidosis. AB - Systemic amyloidosis commonly affects the heart. Indeed, cardiac symptoms may be the first clinical indicator of underlying amyloid deposition. Using two case studies, this article reviews the latest evidence regarding cardiac amyloidosis. The diagnosis of cardiac involvement can be established through imaging with echocardiography and magnetic resonance. Supportive evidence may be gained from biochemical markers such as serum N-terminal probrain natriuretic peptide (NT proBNP). The main clinical consequences of amyloid deposition are cardiac failure and rhythm disturbances. Attempts to cure the underlying disease process with chemotherapy and/or cardiac and/or liver transplantation have had variable results. Stem-cell transplantation is associated with significant mortality in the context of cardiac involvement. Although newer therapeutic agents are emerging, the overall outlook at this time remains poor. PMID- 16268337 TI - A shared agenda: doctors and managers. PMID- 16268338 TI - Management of blackouts and misdiagnosis of epilepsy and falls. PMID- 16268339 TI - Facing an epidemic of chronic kidney disease. PMID- 16268340 TI - Munchausen's syndrome. PMID- 16268341 TI - A survey of continuing professional development (CPD) in palliative medicine. PMID- 16268342 TI - Hands, foot and mouth disease. PMID- 16268343 TI - [Advances of research on medical image fusion]. AB - This paper analyzes the present situation and focuses of medical image fusion and especially places emphasis on the developing trend of intelligent image fusion and comachine image fusion technologies. PMID- 16268344 TI - [Present situation and prospects of artificial heart pumps in Jiangsu University]. AB - Since 1995, four different types of artificial heart pumps and artificial valvo pumps have been developed in Jiangsu University of China. Three types of heart pumps and valvo-pumps have been applied in animal experiments in University Texas, Medical Branch, USA and in Zhenjiang No.1 People's Hospital of China. The recently-developed UJS-IV pump is a totally implantable trans-ventricular and cross-valvular pump for emergercy treatments. PMID- 16268346 TI - [Preliminary assessment of osteoporosis in vivo MR image analysis]. AB - This paper presents a non-invasive and non-ionizing radiation method for assessment of osteoporosis by analyzing in vivo MR images. Texture features (entropy, coarse, etc...) derived from co-occurrence matrix and neighborhood graystone difference matrix are obtained. These features are significantly different between patients and control subjects. Then the grayscale MR image is transformed to the binary image, The shape and topology features (area, skeleton length, euler number, etc. ) obtained from the binary image show too significant differences between patients and control subjects. PMID- 16268345 TI - [A simulation design of a one-way micro valve for the micro engineering capsule]. AB - This paper proposes a one-way micro valve with a simple structure and a simulation design for the engineering capsule. We have now got its design parameter selection method and its mechanic characteristic from experiments. PMID- 16268348 TI - [Development of a new system for head and neck stereotactic conformal radiotherapy]. AB - This paper introduces the constitution and fabrication of the new head and neck stereotactic conformal radiotherapy system. It remedies the shortages in the head and neck stereotactic conformal radiotherapy at present and it is deserved to be popularized in clinical applications. PMID- 16268349 TI - [Software and hardware design for the temperature control system of quantitative polymerase chain reaction]. AB - A temperature control system for quantitive polymerase chain reaction (PCR) is presented in the paper with both software and hardware configuration. The performance of the control system has been improved by optimizing the software and hardware design according to the system's properties. The control system has been proven to have a good repeatability and reliability as well as high control precision. PMID- 16268350 TI - [Cardiovascular circulation feedback control treatment instrument]. AB - The cardiovascular circulation feedback control treatment instrument (CFCTI) is an automatic feedback control treatment system, which has the function of monitoring, alarming, trouble self-diagnosis and testing on the line in the closed loop. The instrument is designed based on the successful clinical experiences and the data are inputted into the computer in real-time through a pressure sensor and A/D card. User interface window is set up for the doctor's choosing different medicine. The orders are outputted to control the dose of medicine through the transfusion system. The response to medicine is updated continually. CFCTI can avoid the man-made errors and the long interval of sampling. Its reliability and accuracy in rescuing the critical patients are much higher than the traditional methods. PMID- 16268347 TI - [Development of external therapeutical ultrasound systems (ETUS) for acute myocardial infarction]. AB - This paper introduces the principle and structure of the ETUS system for acute myocardial infarction. This system is an assistant treatment tool with ultrasound waves acting from the external on the heart, speeding thrombolytic drug's permeation into the thrombi for a good curative effect. PMID- 16268351 TI - [Development of a mobile digital hydraulic extracorporeal heart compression machine]. AB - This article introduces the working principle, the structural design of a mobile digital hydraulic extracorporeal heart compression machine and its trial result on the human model. The result shows that the machine which has the advantages of easy operation, fast effectiveness, safety, line display and agile adjustment, is an ideal medical device for patients with cardiac arrest and is of great social benefit and great market expectations. PMID- 16268352 TI - [The EICP's development for clean operation rooms]. AB - This paper introduces the principium and application of the embedded intelligence control platform (EICP) in the clean operating room in our hospital. It can be a master of automatic control for air decontamination, temperature, humidity, lighting lamps, shadowless lamp, etc.. PMID- 16268353 TI - [Development of ZM-1 tissue microarrayer]. AB - ZM-1 tissue microarrayer designed by our group is manufactured in stainless steel and brass. It features an easier and faster preparation for tissue microarrays. By means of it, a group of biopsy needles are used to punch the donor tissue specimens respectively, and all the needles with the punched specimen cylinders are arranged into the array-board, where small holes have been digged to fit the needles. All the specimen cylinders arraying and the tissue microarray block's shaping are finished simultaneously. ZM-1 tissue microarrayer with a lower cost of manufacture, is capable of preparing the tissue microarrays conveniently, efficiently and quality-controllably. PMID- 16268354 TI - [The inflatable penile prosthesis made in China for treatment of erectile dysfunction]. AB - Erectile dysfunction (ED) is a common male sexual dysfunction. Some ED patients who do not respond to any nonsurgical therapy will eventually be candidate for penile prostheses. This paper introduces the characteristics and application of inflatable penile prosthesis made in China. PMID- 16268355 TI - [The design of an electromyogram-guided treatment instrument]. AB - The text introduces an electromyogram-guided treatment instrument with simple operation and lower cost, and it is easy to find the lesion muscle. Its clinical tests have shown a satisfying result. PMID- 16268356 TI - [Target volume calculation methods based on PET-CT images]. AB - The applications of PET-CT have developed from qualitative analysis to quantitative analysis. Target volume is important for tumor biological volume defining, tumor isotope therapy, organ function evaluation, acceptor affinity calculation, and pharmaceutical metabolic kinetics. Many factors work on the target volume calculation, such as PET image acquisition mode, scatter correction, attenuation correction, reconstruction method, image display mode, positron pharmacy. The commonly-used methods of target volume calculation are background-threshold, max threshold, and background-max threshold. In this article we will discuss about the methods of target volume calculation. PMID- 16268357 TI - [Confirmation of pipetting performances for fully-open automatic biochemistry analyzers]. AB - This paper introduces a kind of evaluation method in pipetting performance on new fully automated biochemistry analyzers by experiments. The performance of sample pipetting volume is confirmed by dye dilution method, the performance of reagent pipetting volume and dummy volume is done by weighing method. Meanwhile, researches and comparative researches on dummy volumes in different conditions have been made, providing valuable reference for clinical applications of automatic biochemistry analyzers. PMID- 16268358 TI - [Application of "P" control chart to quality control of medical devices]. AB - This article describes the opportune time, method and procedures of "P" control chart's establishment during product manufacturing. "P" control chart of statistical data is the most commonly used in data analysis and the quality control system. PMID- 16268359 TI - [Research on telerobotic laparoscopic surgery and its applications]. AB - A general review is here presented on the development, composition, existing problems and prospects of telerobotic laparoscopic surgery systems, based on the related literatures and informations in recent years. PMID- 16268361 TI - [Research on quality system management of software manufacturers for medical devices]. AB - Based on the investigation and analysis of the current situation of medical devices' software manufacturers an exploration of new administrative modalities for the enterprises is presented in the paper. PMID- 16268360 TI - [The research actualities and developing trend of nitric oxide (NO) inhalation systems]. AB - This paper discusses the limitations of current NO inhalation systems, based on the research in collocation of NO, inspection of NO/NO2 and synchronous working of NO inhalation systems with ventilators. And then, the developing trend of NO inhalation systems is put forward here too. PMID- 16268362 TI - [The testing methods of the linearity errors and the cross contamination for clinical analyzers]. AB - The measuring methods of the linearity errors and the cross contamination are very hard to grasp in testing the clinical analyzer's specifications. In this paper, the measuring steps and the data processing methods are introduced in detail in combination with the living examples and taking the form of the lists so as to help the technical personnels to understand the verification regulation of clinical analyzers correctly and to carry out the technical supervision effectively. PMID- 16268363 TI - [The functional integrity of imaging diagnostic equipments]. AB - On the basis of introduction and analysis of three new medical imaging equipments, the article discusses the superiority of the function integrity that is a new concept, and its application in the development of medical equipments in 21st century. PMID- 16268365 TI - [Medical devices' management and medical safety]. AB - This article presents some suggestions about how to strengthen the management of medical devices so as to remove the hidden perils and risks of medical safety. PMID- 16268364 TI - [A discussion on requirements and technical specifications while procuring and deploying a ventilator]. AB - Based on the practical work, this paper discusses the requirements and technical specifications we should pay attention to while procuring and deploying a ventilator. PMID- 16268366 TI - [Experiences in maintenance and repair cost control of medical equipments]. AB - This paper introduces methods to control the cost of maintenance and repair for medical equipments through service team training, service contract control, system establishment and outside service resources, etc.. PMID- 16268367 TI - Sleep habits and accident risk among truck drivers: a cross-sectional study in Argentina. PMID- 16268368 TI - Sleep and suicidality: do sleep disturbances predict suicide risk? PMID- 16268370 TI - Insomnia: a dangerous condition but not a killer? PMID- 16268369 TI - How many children with ADHD have sleep apnea or periodic leg movements on polysomnography? PMID- 16268371 TI - Insomnia state of the science: an evolutionary, evidence-based assessment. PMID- 16268372 TI - The diagnosis and treatment of chronic insomnia in adults. PMID- 16268373 TI - National Institutes of Health State of the Science Conference statement on Manifestations and Management of Chronic Insomnia in Adults, June 13-15, 2005. PMID- 16268374 TI - The neural basis of the psychomotor vigilance task. AB - STUDY OBJECTIVE: To identify brain regions underlying the fastest and slowest reaction times on the Psychomotor Vigilance task (PVT) under well-rested conditions, as well as brain regions related to particularly poor performance after sleep deprivation. DESIGN: Subjects took the PVT twice while undergoing functional magnetic resonance imaging: once 12 hours after waking from a normal night of sleep and once after 36 hours of total sleep deprivation (TSD). Session order was counterbalanced. SETTING: UCSD J. Christian Gillin Laboratory for Sleep and Chronobiology (the sleep core of the General Clinical Research Center) and UCSD Magnetic Resonance Institute. PATIENTS OR PARTICIPANTS: Twenty right-handed healthy adults (8 women; age = 27.4 +/- 6.7 years; education = 15.6 +/- 1.5 years). MEASUREMENTS AND RESULTS: After a normal night of sleep, optimal performance was related to greater cerebral responses within a cortical sustained attention network and the cortical and subcortical motor systems. Slow reaction times, particularly after TSD, were associated with greater activity in the "default mode network" consisting of frontal and posterior midline regions. CONCLUSIONS: Optimal performance on the PVT appears to rely on activation both within the sustained attention system and within the motor system. Poor performance following TSD may result from a disengagement from the task and related inattention, and brain regions responsible for this localize within midline structures shown to be involved in the brain's "default mode." Finally, particularly poor performance after TSD may elicit a subsequent attentional recovery that manifests as greater activation within the same regions normally responsible for fast reaction times. PMID- 16268375 TI - Ferritin levels in the cerebrospinal fluid and restless legs syndrome: effects of different clinical phenotypes. AB - STUDY OBJECTIVE: To determine whether patients with restless legs syndrome (RLS) and controls differ in regard to levels of ferritin and transferrin in the cerebrospinal fluid (CSF) when samples are collected at night, to determine whether patients with early-onset and late-onset RLS show a different outcome for CSF values, and to determine whether the CSF ferritin level correlates with disease severity. DESIGN: Collection of CSF and plasma; assessment of disease severity using objective (periodic limb movements) and subjective (Johns Hopkins Restless Legs Severity Scale) measures of severity. SETTING: General Clinical Research Center. PARTICIPANTS: Thirty subjects with idiopathic RLS (15 early- and 15 late-onset RLS) and 22 age- and sex-matched controls. INTERVENTION: N/A. RESULTS: Nighttime CSF ferritin levels were lower in the total RLS group compared with controls. Further assessment found that the early-onset (less than 45 years of age) but not the late-onset (greater than or equal to 45 years of age) RLS group had significantly lower CSF ferritin levels compared with controls. There was a strong correlation between the age of symptom onset and CSF ferritin values (r = 0.64): the earlier the age, the lower the ferritin level. A regression analysis showed that both sex and RLS subtype had significant effects on the CSF ferritin level, with women with early-onset RLS having substantial lower values than men with late-onset RLS. A comparison between these nighttime CSF values and previously published daytime samples suggests that diurnal changes may have effects on the findings. CONCLUSIONS: This study is distinct in showing that the degree of the CSF-ferritin effect is best defined by the clinical phenotypes of sex and age of symptom onset and by the time of day that samples are collected. PMID- 16268376 TI - Brain potentials before and after rapid eye movements: an electrophysiological approach to dreaming in REM sleep. AB - STUDY OBJECTIVE: This study examined hypotheses regarding dreaming in rapid eye movement (REM) sleep by comparing brain potentials related to rapid eye movements in REM sleep with those in wakefulness. DESIGN: Within participants. SETTING: Data were collected in a sleep laboratory. PARTICIPANTS: Fifteen healthy university students who reported having dreams frequently. MEASUREMENTS AND RESULTS: Rapid eye movements in REM sleep were recorded during natural nocturnal sleep. Saccades in wakefulness were recorded during a self-paced visual search task. The presaccadic negativity before and the lambda response after eye movements were examined. It was assumed that the presaccadic negativity reflects voluntary readiness activity before eye movements, and the lambda response reflects visual information processing after saccades in wakefulness. Brain potentials were averaged, time-locked to the onset and offset of eye movements for the presaccadic negativity and the lambda response, respectively. In wakefulness, the presaccadic negativity occurred at the centroparietal site. However, no presaccadic negativity was found during REM sleep. Lambda-like responses (P1r, P2r) were observed in REM sleep over the parietooccipital site, as were the lambda responses (P1, P2) in wakefulness. CONCLUSIONS: The finding that rapid eye movements are initiated without preparation but elicit some neural activity in the cortical visual area suggests that rapid eye movements may trigger dream images. PMID- 16268377 TI - Partial REM-sleep deprivation increases the dream-like quality of mentation from REM sleep and sleep onset. AB - STUDY OBJECTIVES: Sleep onset (SO) is cognitively and physiologically similar to rapid eye movement (REM) sleep, supporting the notion that REM sleep-related processes are 'covertly' active at this time. The objective was to determine if SO mentation is sensitive to REM sleep deprivation. DESIGN: Two-group cross sectional design; sleep recordings for 3 nights. SETTING: Standard sleep laboratory with 24-channel polysomnography recording. PARTICIPANTS: Fourteen female, 13 male healthy volunteers (18-41 yrs, mean=24.8 +/- 6.07). INTERVENTIONS: On Night 2, half were and half were not partially REM sleep deprived (REMD), recalled REM mentation, and rated it for dream-like quality (DLQ), sleepiness, and sensory attributes. On Night 3, all were awakened from SO substages 4 and 5 for mentation reports and further ratings. REMD measures were derived from scored sleep tracings. MEASUREMENTS AND RESULTS: REMD produced increases in DLQ for both REM and SO reports (P < .05); DLQ scores were higher for REM than for SO mentation (P < .001). Covarying sleepiness preserved the (REMD) effect but abolished the REM/SO difference. Whereas 2 sensory attributes (presence of self, visual intensity) tended to distinguish the REM-mentation reports of REMD and control subjects, only 1, self-movement, distinguished their SO mentation reports (P < .06). Multiple regression revealed that increased DLQ of both REM and SO mentation was associated with increased sleepiness and decreased REM sleep time on Night 2. CONCLUSIONS: SO mentation responds to REMD much like REM mentation does, a finding consistent with other work supporting the notion of covert REM-sleep processes at SO. DLQ may be mediated by both increases in REM-sleep propensity and a circadian process indexed by sleepiness ratings. PMID- 16268378 TI - Age-related changes in the circadian modulation of sleep-spindle frequency during nap sleep. AB - STUDY OBJECTIVES: Sleep spindles exhibit a clear circadian modulation in healthy younger people. During the biological night (when melatonin is secreted), spindle density and spindle amplitude are high and spindle frequency and its variability are low, as compared with the biological day. We investigated whether this circadian modulation of spindle characteristics changes with age. DESIGN: A 40 hour multiple-nap paradigm under constant-routine conditions SETTING: Chronobiology Laboratory, University Psychiatric Hospitals, Basel, Switzerland PARTICIPANTS: Seventeen younger (20-31 years) and 15 older (57-74 years) volunteers. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: Whereas the circadian modulation of spindle density, amplitude, duration, and intraspindle frequency variability was not greatly affected by age, we found significant changes in the circadian modulation of spindle frequency. The pronounced circadian modulation of spindle frequency in younger, but not older, subjects was phase locked with the circadian rhythm in melatonin secretion. In the latter, circadian modulation was attenuated and tended to be advanced with respect to the timing of melatonin secretion. There was no difference between age groups in the phase of the sleep wake cycle or that of melatonin, nor did the phase angle between them differ. Although changes in the circadian modulation of spindle frequency in older subjects were accompanied by reduced amplitude in the sleep consolidation profile, there was no significant correlation between spindle frequency and sleep consolidation. CONCLUSION: This multiple-nap protocol under constant-routine conditions revealed an age-dependent weaker coupling of the circadian rhythms of spindle frequency and sleep propensity to the circadian rhythm of melatonin secretion. PMID- 16268379 TI - Sleep habits and accident risk among truck drivers: a cross-sectional study in Argentina. AB - Road accidents are a major cause of death, and sleep deprivation affects driving skills. We conducted a cross-sectional study to evaluate sleep habits and accident risk in long-haul truck drivers in Buenos Aires, Argentina. Questionnaires regarding sleep habits, snoring, and daytime sleepiness were administered, and a limited physical examination was performed. We obtained 738 complete answers (response rate 85%). Mean sleep hours during working days was 3.76 (SD 2.40). Mean driving hours was 15.9 (SD 5.60) per day. Frequent sleepiness while driving was reported by 43.7% of responders. Sleepiness while driving was associated with Epworth Sleepiness Scale values >10 (odds ratio 1.85, 95% confidence interval = 1.20-2.85). Snoring was reportea by 71% of drivers and was frequent in 43.8%. Snoring more than 3 times a week (odds ratio 1.73, 95% confidence interval = 1.23-2.44), sleepiness while driving (OR 1.92, 95% confidence interval = 1.08-1.96), and Epworth Sleepiness Scale score > 10 (odds ratio 2.53, 95% confidence interval = 1.61-3.97) were independently associated with reporting of accidents or near accidents. Sleep deprivation and long driving shifts were prevalent in our study. Accident risk was associated with frequent snoring, daytime sleepiness, and reporting of sleepiness at the wheel. This study highlights the need of improving working conditions in this highly exposed population. PMID- 16268380 TI - Differential effects of sleep deprivation on saccadic eye movements. AB - STUDY OBJECTIVES: This study was designed to show the influence of sleep deprivation on different types of saccadic eye movements. DESIGN: Performance of saccadic eye movements was compared after normal sleep and sleep deprivation in a randomized, within-subjects paradigm. Parameters of voluntary and reflexive saccades were measured before and after experimental nights and after a night of recovery sleep. Additionally, subjects spent 1 adaptation night in the laboratory before the experiments. SETTING: Experiments took place under controlled laboratory conditions. PARTICIPANTS: Fifteen healthy male volunteers (aged 19-30 years). INTERVENTIONS: Each subject participated in 1 night of sleep deprivation followed by a night of recovery sleep and, on another occasion, in 2 successive nights of undisturbed sleep. MEASUREMENTS AND RESULTS: Horizontal prosaccades, antisaccades, and memory-guided saccades were recorded by means of electrooculography. They were analysed semiautomatically with respect to accuracy, peak velocity, and latency. Peak velocity was significantly reduced in all saccade tasks after 1 night of sleep deprivation but recovered after another night of sleep. Latency was prolonged after sleep deprivation only for memory guided saccades; accuracy showed a decrease after 1 night without sleep only for prosaccades. CONCLUSIONS: Sleep deprivation has a general impairing effect on the peak velocity of saccades, reflecting possible dysfunction at the level of the brainstem reticular formation. Deficits of accuracy and latency point to dysfunction of specific brain sites such as the supplementary eye field and cerebellum, whereas the cardinal functions of the frontal and parietal eye fields were not affected. These results suggest the possibility of measuring fatigue by means of saccadic parameters, especially saccadic peak velocity. PMID- 16268381 TI - Comparison of nasal pressure transducer and thermistor for detection of respiratory events during polysomnography in children. AB - STUDY OBJECTIVES: The results of small studies have suggested that a nasal cannula pressure transducer has a higher sensitivity than a thermistor in detecting hypopneas and diagnosing sleep-disordered breathing in both adults and children. We compared a thermistor alone, and in conjunction with a pressure transducer, for detection of sleep-disordered breathing in children during in home polysomnography. DESIGN: Retrospective analysis of a subsample of a prospective cohort study. SETTING: Students attending elementary school in the Tucson Unified School District. PARTICIPANTS: A subsample of the Tucson Children's Assessment of Sleep Apnea study population. MEASUREMENTS AND RESULTS: Polysomnographic recordings of 40 children (24 girls and 16 boys, mean age 9.2 +/ 1.7 years; range 6-11 years) were analyzed to compare the detection of sleep disordered breathing events by 2 different methods of measuring airflow: thermistor alone and thermistor with nasal-cannula pressure transducer (transducer) used simultaneously. The transducer detected all the respiratory events detected by the thermistor, but the thermistor detected only 84% of the transducer-defined events. Consequently, the transducer-derived mean respiratory disturbance index was higher than that detected by the thermistor (7.0 +/- 3.8 vs 5.9 +/- 3.4, P < .001). The bias error between transducer respiratory disturbance index and thermistor respiratory disturbance index on a Bland-Altman plot was 1.08 (95% confidence interval, 0.8 - 1.4). There was good agreement between the thermistor and the transducer for making the diagnosis of sleep apnea using a cutoff of a respiratory disturbance index greater than 5 (kappa = 0.69). The quality of the tracings with the transducer was comparable to that of the thermistor, but the transducer dislodged more frequently. CONCLUSION: The use of a nasal transducer in conjunction with a thermistor was more sensitive than the thermistor alone in detecting sleep-disordered breathing in children during unattended polysomnography. PMID- 16268383 TI - Suicidality and sleep disturbances. AB - STUDY OBJECTIVES: A growing body of research indicates that sleep disturbances may be specifically linked to suicidal behaviors. It remains unclear, however, whether this link is largely explained by depressive symptoms. The present study investigated the relationship between suicidality, depression, and sleep complaints in a clinical outpatient setting. DESIGN AND SETTING: Upon admission, 176 outpatients completed measures on sleep disturbances, suicidal symptoms, and depression. Several sleep disturbances were evaluated with regard to suicidal ideation, including insomnia, nightmares, and sleep-related breathing symptoms. MEASUREMENTS AND RESULTS: Regression analyses revealed that insomnia and nightmare symptoms were associated with both depressive symptoms and suicidality. Sleep-related breathing symptoms were associated with depressive symptoms, but did not show an association with suicidal ideation. After controlling for depressive symptoms, only nightmares demonstrated an association with suicidal ideation. This relationship emerged as a nonsignificant trend (P = .06). Nightmares were particularly associated with suicidality among women compared with men. Posthoc analyses revealed that, after controlling for sex and depressive symptoms, nightmare symptoms were significantly associated with suicidality (P = .04). CONCLUSIONS: Although insomnia and nightmares were significantly associated with depressive and suicidal symptoms, after controlling for additional variables, such as depression and sex, only nightmares remained associated with suicidality. This association was slightly stronger among women compared with men. PMID- 16268382 TI - Polysomnographic predictors of blood pressure and hypertension: is one index best? AB - STUDY OBJECTIVES: Numerous indexes derived from polysomnography are available to characterize sleep-disordered breathing, with no consensus over which measures best predict clinical outcomes. This study addresses the relative merits of using alternative polysomnography indexes by characterizing the consistency and strength of the association of each index with blood pressure and hypertension. DESIGN: Cross-sectional analyses of the association of alternative polysomnography indexes with blood pressure and hypertension were performed in construction and validation data sets. Linear and logistic regression models were used to identify the best variable sets. PATIENTS: Data were obtained from 6433 men and women (age 62.9 +/- 11.0 years, 52.8% women) who participated in the Sleep Heart Health Study. RESULTS: In multivariable models, most indexes showed weak linear associations with systolic, with slightly stronger associations for diastolic blood pressure, and the log odds of hypertension. No single index showed consistent superiority over others. Systolic blood pressure, diastolic blood pressure, and hypertension each were associated with distinct sets of polysomnography variables. Slightly more-consistent associations were demonstrated for indexes that included hypopneas that were linked with either a 3% or 4% desaturation level than indexes that did not require hypopneas to have linked desaturation. For indexes that combined apneas and hypopneas, there was no evidence that linking obstructive apneas to desaturation or arousal altered prediction compared with counting all apneas. CONCLUSION: In summary, using a rigorous cross-validation assessment, we did not identify a clear superiority of any single index for blood pressure or hypertension prediction. Detailed analyses of alternative definitions of the respiratory disturbance index support current scoring guidelines, where desaturation criteria are recommended for hypopneas but not apneas. PMID- 16268384 TI - Patients with attention-deficit/hyperactivity disorder without observed apneic episodes in sleep or daytime sleepiness have normal sleep on polysomnography. AB - STUDY OBJECTIVE: There is continuing speculation about the relationship between attention-deficit/hyperactivity disorder (ADHD) and obstructive sleep apnea (OSA) or periodic limb movement disorder (PLMD)/restless legs syndrome. The objective was to determine if a significant portion of children with ADHD diagnosed using DSM-IV criteria have OSA or PLMD. SETTING: Sleep disorders centers in a private practice setting and a hospital setting. PARTICIPANTS: Children aged 6 to 14 years with ADHD were enrolled. Patients with snoring were not excluded. Although patients with snoring plus either observed apneic episodes in sleep or excessive daytime sleepiness were to be excluded, as were patients with restless legs at night, only 1 subject actually had to be excluded because of these criteria. MEASUREMENTS AND RESULTS: Forty children were evaluated with a polysomnogram. A respiratory disturbance index cut-off of more than 5 per hour of sleep was used to diagnose OSA, and a periodic limb movement (with arousal) index cut-off of 5 or more per hour of sleep was used to diagnose PLMD. The Attention Deficit/Hyperactivity Disorder Rating Scale-IV-Parent Version: Investigator Administered and Scored was used to determine severity of inattentive, hyperactive, and total ADHD symptoms. Except for a somewhat longer rapid eye movement sleep latency and decreased percentage of rapid eye movement sleep, polysomnography was essentially normal. No patient had OSA or PLMD on polysomnography. CONCLUSIONS: OSA or PLMD is not a common underlying disorder or etiologic factor in patients who meet the criteria for ADHD. In the absence of symptoms suggesting a primary sleep disorder, such as snoring with observed apneic episodes in sleep or daytime sleepiness or restless legs, polysomnographic evaluation does not seem indicated in patients with ADHD. PMID- 16268385 TI - An electrocardiogram-based technique to assess cardiopulmonary coupling during sleep. AB - STUDY OBJECTIVES: To evaluate a new automated measure of cardiopulmonary coupling during sleep using a single-lead electrocardiographic signal. DESIGN: Using training and test datasets of 35 polysomnograms each, we assessed the correlations of an electrocardiogram-based measure of cardiopulmonary interactions with respect to standard sleep staging, as well as to the cyclic alternating pattern classification. The pattern of coupling in 15 healthy individuals was also assessed. SETTING: American Academy of Sleep Medicine Accredited Sleep Disorders Center. INTERVENTIONS: None. MEASUREMENTS AND RESULTS: From a continuous, single-lead electrocardiogram, we extracted both the normal-to normal sinus interbeat interval series and a corresponding electrocardiogram derived respiration signal. Employing Fourier-based techniques, the product of the coherence and cross-power of these 2 simultaneous signals was used to generate a spectrographic representation of cardiopulmonary coupling dynamics during sleep. This technique shows that non-rapid eye movement sleep in adults demonstrates spontaneous abrupt transitions between high- and low-frequency cardiopulmonary coupling regimes, which have characteristic electroencephalogram, respiratory, and heart-rate variability signatures in both health and disease. Using the kappa statistic, agreement with standard sleep staging was poor (training set 62.7%, test set 43.9%) but higher with cyclic alternating pattern scoring (training set 74%, test set 77.3%). CONCLUSIONS: A sleep spectrogram derived from information in a single-lead electrocardiogram can be used to dynamically track cardiopulmonary interactions. The 2 distinct (bimodal) regimes demonstrate a closer relationship with visual cyclic alternating pattern and non cyclic alternating pattern states than with standard sleep stages. This technique may provide a complementary approach to the conventional characterization of graded non-rapid eye movement sleep stages. PMID- 16268386 TI - The use of stimulants to modify performance during sleep loss: a review by the sleep deprivation and Stimulant Task Force of the American Academy of Sleep Medicine. PMID- 16268387 TI - Atomoxetine also effective in patients suffering from narcolepsy? PMID- 16268388 TI - "Augmentation" of augmentation index studies in obstructive sleep apnea. PMID- 16268389 TI - HLA DQB1*0602 positive narcoleptic subjects with cataplexy have CSF lgG reactive to rat hypothalamic protein extract. PMID- 16268390 TI - Water treeing in simplified dentin adhesives--deja vu? PMID- 16268391 TI - Clinical evaluation of a flowable resin composite and flowable compomer for preventive resin restorations. AB - This clinical study evaluated the retention and caries protection of a flowable resin composite (Flow Line) and a flowable compomer (Dyract Flow) used in preventive resin restorations as compared to the conventional preventive resin technique which uses a resin composite (Brilliant) and a sealant (Concise). This study observed 205 permanent molars with small carious cavities less than 1.5 mm in width, which were obtained from 165 children aged 7 to 15 years. Flowable resin composite was used to treat 75 teeth, and 71 teeth were treated with flowable compomer in both cavities and caries-free fissures. For the control group, 59 teeth were treated with resin composite in cavities and sealant in caries-free fissures. The teeth were evaluated at 3, 6, 12, 18 and 24-month intervals. After three months, all 205 treated teeth were completely intact. After six months, 66 of the 71 teeth treated with flowable resin composite and 65 of the 70 teeth treated with flowable compomer were complete, compared to 57 of the 58 teeth treated with the conventional preventive resin technique. After 12 months, 60 of the 67 teeth treated with flowable resin composite and 61 of the 67 teeth treated with flowable compomer were complete, compared to 51 of the 55 teeth treated with the conventional preventive resin technique. After 18 months, 53 of the 61 teeth treated with flowable resin composite and 54 of the 62 teeth treated with flowable compomer were complete, compared to 47 of the 53 teeth treated with the conventional preventive resin technique. After 24 months, 49 of the 58 teeth treated with flowable resin composite and 45 of the 57 teeth treated with flowable compomer were complete, compared to 42 of the 52 teeth treated with the conventional preventive resin technique. There were no statistically significant differences in retention rates among all groups after 3, 6, 12, 18 or 24-months (p>0.05). One tooth treated with flowable resin composite and one tooth treated with flowable compomer developed caries after 18 and 24 months, respectively, resulting from partial loss at "caries-free fissures." Five teeth developed caries in the conventional preventive resin group; one after 12 months, two after 18 months and one after 24 months, due to loss at cavities. The final caries occurred after 24 months, resulting from partial loss at "caries-free fissures." The differences in caries development among the three groups were not statistically significant (p>0.05). This study suggested that flowable resin composite and flowable compomer could be used for preventive resin restorations. Meanwhile, a vigilant recall should be followed-up due to the risk of failure for flowable materials in "caries-free" fissures. The repair should be performed immediately, in case the preventive resin restoration develops a fracture or loss. PMID- 16268392 TI - A clinical evaluation of bleaching using whitening wraps and strips. AB - This study evaluated the degree of color change of teeth and the sensitivities of teeth and gums in an in vivo study. Ranir Whitening Wraps (WW2) and Crest Whitestrips Premium (WP2) were used twice a day and Ranir Whitening Wraps (WW1) were used once a day. Color evaluations occurred at baseline, after five and seven-day use of bleaching agent and 14 days post-bleaching. Color change was evaluated objectively and subjectively. Sensitivity evaluations were also accomplished. Seventy-six of the 78 subjects enrolled completed the study. All three products significantly lightened teeth. WW2 lightened more than WP2 and WW1 in L*, a*, b*, E and shade guide value. WP2 lightened more than WW1 in a*, b*, E and shade guide value. There was no difference in tooth sensitivity, but WW1 and WP2 caused less gingival sensitivity than WW2. The mean age of smokers was seven years younger than nonsmokers who qualified. PMID- 16268393 TI - Surface finish produced on three resin composites by new polishing systems. AB - This study evaluated the surface finish of three direct resin composites polished with three different systems. Disk-shaped specimens (n=16 per material; phi=8.0 mm x h=2.0 mm) were formed in a stainless steel mold by packing uncured material, either a hybrid composite (Z250, 3M ESPE) or two micro-hybrid composites (Point 4, Kerr; Esthet-X, Dentsply), and light-cured from the top and the bottom surfaces with a light-emitting diode (LED) curing unit (NRG, Dentsply). After storing the specimens in deionized water at 37 degrees C for seven days, one side of each specimen was finished through 1200-grit SiC abrasive (Buehler). Five specimens of each resin composite were randomly assigned to one of the three polishing systems (Identoflex, Kerr; Pogo, Dentsply; Sof-Lex, 3M ESPE). Manufacturers' instructions were followed during the polishing procedures. The average surface roughness (Ra) was determined by generating tracings across the polished surface of each disk using a scanning profilometer (Surfanalyzer System 5000, Federal Products Co). The results were analyzed by Kruskal-Wallis and Mann and Whitney tests (p < or = 0.05). The smoothest surfaces were produced with the celluloid strip (control group) on all the resin composites tested. The aluminum oxide disks (Sof-Lex) produced a statistically equivalent surface finish (Ra) on the three resin composites. The lowest mean roughness values were recorded with diamond micropolisher disks (PoGo) on the hybrid composite (Z250). Overall, the two new polishing systems, Identoflex and PoGo, created a comparable surface finish to that produced by the Sof-Lex system on all three resin composites. PMID- 16268394 TI - Influence of vision on the evaluation of marginal discrepancies in restorations. AB - This study investigated the influence of visual inspection in the detection and discrimination between principle types of marginal discrepancies in restorations. Using devices simulating vertical steps, horizontal gaps and the combination of a vertical step and horizontal gap at the margin of a restoration, and explorers with four different tip diameters, 10 experienced dental faculty members were asked to identify discrepancies and the boundary between Alpha (excellent) and Bravo (clinically acceptable) marginal adaptation ratings under three different visual conditions--with and without visual inspection and visual inspection aided with binocular loupes. A significant correlation was found to exist between explorer tip diameter and the Alpha/Bravo boundary for horizontal gaps, but not for vertical steps. There was no significant difference in the detection of the Alpha/Bravo boundary for the three visual conditions. It was concluded that visual inspection aided and unaided with loupes had no significant effect on the evaluation of simulated marginal discrepancies. These findings highlight the importance of the traditional dental explorer, in the absence of a more discriminatory devices in the assessment of marginal discrepancies in restorations. PMID- 16268395 TI - Effect of polymerization modes and resin composite on the temperature rise of human dentin of different thicknesses: an in vitro study. AB - This in vitro study evaluated the effect of different polymerization modes and the presence of resin composite on the temperature rise (TR) in human dentin of different thicknesses. For this purpose, 90 specimens were assigned to 30 groups (n=3): five polymerization modes (1-conventional; 2-soft-start; 3-high intensity; 4-ramp cure: progressive and high intensity; 5-high intensity with the tip of the light cure at a distance of 1.3 cm for 10 seconds and the tip leaned in the sample); two levels of resin composite presence (absence or presence of resin composite) and three dentin thicknesses (1, 2, 3 mm). During polymerization, temperature was measured by a digital laser thermometer (CMSS2000-SL/SKF). Three way ANOVA and Tukey tests were performed. There were statistical differences in TR among polymerization modes, presence of resin composite and dentin thicknesses. Within the limits of this study, it can be concluded that 1) conventional and high intensity polymerization modes presented lower TR means, and it was statistically different from soft start, distanced tip and ramp curing polymerization modes; 2) the presence of resin composite showed a statistically significant reduction TR means and 3) the thicker the dentin, the less the temperature rise. PMID- 16268396 TI - The effect of 10% carbamide peroxide, carbopol and/or glycerin on enamel and dentin microhardness. AB - This study evaluated the effects of 10% carbamide peroxide, carbopol and glycerin and their associations on microhardness over time on enamel and dentin. Eight treatment agents were evaluated: a commercial bleaching agent containing 10% carbamide peroxide (Opalescence 10% Ultradent), 10% carbamide peroxide, carbopol, glycerin, 10% carbamide peroxide + carbopol, 10% carbamide peroxide + glycerin, carbopol + glycerin and 10% carbamide peroxide + carbopol + glycerin. Three hundred and twenty human dental fragments, 80 sound enamel fragments (SE), 80 demineralized enamel fragments (DE), 80 sound dentin fragments (SD) and 80 demineralized dentin (DD) fragments, were exposed to the treatment agents (n=10). These agents were applied onto the surface of the fragments eight hours a day for 42 days. After eight hours, they were washed from the dental fragment surfaces after five back-and-forth movements with a soft bristle toothbrush under distilled and deionized running water. During the remaining time (16 hours per day), the fragments were kept in individual vials in artificial saliva. After the 42-day treatment period, the specimens were kept individually in artificial saliva for 14 days. Knoop microhardness measurements were performed at baseline, after eight hours, and 7, 14, 21, 28, 35 and 42 days, and 7 and 14 days post treatment (corresponding to 49 and 56 days after the initial treatment agent applications). The non-parametric Kruskal-Wallis analysis showed significant differences among the agents at each time interval, except at baseline for sound and demineralized enamel and dentin. For SE, SD and DD, there was a decrease in microhardness values during treatment with all agents. There was a tendency towards lower microhardness values after treatment with carbopol and its associations for sound tissues. DD showed low microhardness values during and after treatment with CP and its associations. For DE, there was an increase in microhardness values during treatment with all agents and in the post-treatment phase. The baseline microhardness values were not recovered during the 14-day post-treatment phase. Opalescence 10%, carbamide peroxide, carbopol, glycerin and their associations may change the microhardness of sound and demineralized dental tissues, even in the presence of artificial saliva. PMID- 16268397 TI - Surface roughness of different dental materials before and after simulated toothbrushing in vitro. AB - This study measured the effect of toothbrushing with a slurry of toothpaste on different dental materials that have been optimally polished. Specimens (n=8) of 21 dental materials, 16 resin composites, 1 amalgam and 4 ceramic materials, were subjected to 36,000 cycles (approximately five hours) of circular toothbrushing with a force of 1.7 N and a slurry of toothpaste (RDA 75) in a device for simulated toothbrushing. The unpolished enamel and dentin of extracted anterior teeth were used as a control. The mean roughness (Ra) was measured with an optical sensor (FRT MicroProf) before and after toothbrushing. To compare the roughness of the different materials, ANOVA with a post hoc Tukey B test was applied (p<0.05). Among the resin composites, the hybrid composites showed the greatest increase in mean roughness, while the microfilled composites and the compomer Compoglass F demonstrated the lowest increase. No statistically significant difference in roughness was found before and after simulated toothbrushing for the Amalcap amalgam, Esthet-X resin composite, TPH Spectrum resin composite, d.SIGN ceramic and the experimental ceramic. The other ceramic materials and dentin and enamel specimens showed a statistically significant decrease in mean roughness after simulated toothbrushing. Glazed Empress demonstrated a statistically significant higher initial roughness than polished Empress. For resin composites, no correlation was found between the mean particle size and mean roughness after simulated toothbrushing. PMID- 16268398 TI - Evaluation of the adhesion of fiber posts to intraradicular dentin. AB - The interfacial strength and ultrastructure of a total-etch, self-etch and self adhesive resin cement used to lute endodontic glass fiber posts (FRC Postec, Ivoclar-Vivadent) was assessed with the "thin-slice" push-out test and transmission electron microscopy (TEM). The tested adhesive cements were Variolink II (Ivoclar-Vivadent), Panavia 21 (Kuraray Co) and RelyX Unicem (3M ESPE). In each group, seven posted roots were used for push-out tests and two were processed for TEM observations. The interfacial strength achieved by Variolink II (10.18 +/- 2.89 MPa) was significantly higher than Panavia (5.04 +/- 2.81 MPa) and RelyX Unicem (5.01 +/- 2.63 MPa), which were comparable to each other. TEM micrographs of the interface between Variolink II and intraradicular dentin revealed that the smear layer was totally removed and an 8-10 micron thick hybrid layer was formed. In the other group specimens, the smear layer was not completely dissolved and smear plugs were retained. Gaps were present between the hybridized complex and the adhesive layer in the Panavia 21 specimens and between the smear layer and underlying root dentin in the RelyX Unicem specimens. Interfacial strengths and microscopic findings were in agreement and indicated that the bonding potential of the total-etch resin cement was greater. The acidic resin monomers responsible for substrate conditioning in Panavia 21 and RelyX Unicem appeared unable to effectively remove the thick smear layer created on root dentin during post space preparation. PMID- 16268399 TI - Inhibitory activity of glass-ionomer cements on cariogenic bacteria. AB - This study evaluated the antibacterial activity of the glass-ionomer cements Vitrebond (3M ESPE), Ketac Molar (3M ESPE) and Fuji IX (GC America) against S mutans, S sobrinus, L acidophilus and A viscosus, using the agar diffusion test. Inocula were obtained by the seed of indicators cultures in BHI broth incubated at 37 degrees C for 24 hours. Base layers containing 15 mL of BHI agar and 300 microL of each bacteria suspension were prepared in Petri dishes. Six wells measuring 4 mm in diameter were made in each plate and completely filled with one of the testing materials. A 0.2% chlorhexidine solution applied in round filter papers was used as control. Tests were performed 12 times for each material and bacteria strain. After incubation of the plates at 37 degrees C for 24 hours, the zones of bacterial growth inhibition around the wells were measured. Overall, the results showed the following sequence of antibacterial activity: Vitrebond (despite the activation mode) > 0.2% chlorhexidine > Ketac Molar > Fuji IX, according to Kruskal-Wallis and Mann-Whitney statistical tests. This study confirmed significant antibacterial activity for two conventional glass-ionomers and one resin-modified glass-ionomer material. The resin-modified glass-ionomer cement Vitrebond, regardless of the activation mode, presented the best antibacterial activity against S mutans and S sobrinus. The antibacterial activity against A viscosus for Vitrebond was similar to 0.2% chlorhexidine, while light activation reduced its antibacterial activity against L acidophilus. PMID- 16268400 TI - Influence of light activation on the volumetric change of core foundation resins. AB - A core foundation system is frequently used in endodontically treated teeth that suffer excessive loss of the coronal portion of their structure. The volumetric shrinkage of core foundation resins may create marginal gaps that influence the bonding ability and longevity of a restored tooth. Little is known about how activation conditions of resin core foundation resin pastes affect their volumetric shrinkage. This study evaluated the influence of light intensity and light activation duration on volumetric shrinkage of direct core foundation resins. Two dual- and one light-activated core foundation resin pastes were employed. The material was placed in a Teflon mold 4 mm in diameter and 2 mm in height and extruded into a water filled dilatometer. The specimens were then light activated and the change in height of the meniscus of water was recorded using a charged-coupled device camera. The average volumetric shrinkage of the core foundation resins after 180 seconds ranged from 1.53% to 2.63%. For all materials tested, there was a tendency for increased volumetric shrinkage with increased light activation time and intensity. The results of this study indicate that the volumetric change of core foundation resins is influenced by the time and intensity of light activation. PMID- 16268401 TI - The effect of composite type on microhardness when using quartz-tungsten-halogen (QTH) or LED lights. AB - This study evaluates the Knoop microhardness of resin composites cured with different light-emitting diode (LED) based light curing units (LCU) or with a conventional quartz-tungsten-halogen light (QTH). Ten experimental groups with 10 specimens each were used. The specimens were prepared by placing two light-cured resin composites with similar VITA shade A2-microhybrid Filtek Z250/3M ESPE and microfill Durafil VS/Heraeus Kulzer--in a 2.0 mm-thick disc shaped mold. The specimens were polymerized for 40 seconds with the use of one QTH LCU (Optilux 501/Kerr-Demetron) and four LED LCUs: Elipar FreeLight 1 Cordless LED (3M ESPE), Ultrablue II LED with cord (DMC), Ultrablue III LED cordless (DMC) and LEC 470 I (MM Optics). Knoop microhardness was determined at the top and bottom surfaces of the specimens 24 hours following curing. Microhardness values in the microhybrid resin composite group showed no statistically significant differences when cured with LED FreeLight 1 LCU and QTH LCU (p<0.05). The other LED devices evaluated in the study presented lower microhardness values in both surfaces (p<0.05) when compared to QTH. In the microfill resin composite group, no statistically significant differences were observed among all LCUs evaluated on the bottom surfaces (p<0.05). However, on the top surfaces, QTH presented the highest KHN values, and the LED devices presented similar results when compared with KHN values relative to each other (p<0.05). PMID- 16268402 TI - Two-year color changes of light-cured composites: influence of different light curing units. AB - This study determined color changes in a composite cured with various types of curing units after two years. A hybrid (Clearfil AP-X) composite was cured with a conventional halogen, a high intensity halogen, a plasma arc and a light emitting diode unit. The specimens were stored in light-proof boxes after the curing procedure to avoid further exposure to light and stored in 37 degrees C in 100% humidity. Colorimetric values of the specimens immediately after curing and after two years were measured using a colorimeter. The CIE 1976 L*a*b color system was used to determine color differences. Differences from baseline were calculated as deltaE*ab. Data were analyzed with two-way analysis of variance (p<0.05). The deltaE*ab values varied significantly, depending on the curing unit used. The specimens cured with a plasma arc curing unit induced significantly higher color changes than any other specimen and the color differences were also visually appreciable by the non-skilled operator (deltaE*ab >2.5). The specimens cured with a high intensity halogen curing unit produced the lowest color change; however, there were no statistically significant differences among the color changes of specimens cured with conventional halogen, high intensity halogen and the light emitting diode unit, and the color changes were not clinically relevant (deltaE*ab <2.5). The results of this study suggest that composite materials undergo measurable changes due to curing unit exposure. The specimens cured with a plasma arc light showed the highest color changes as compared to specimens cured with other curing units. PMID- 16268404 TI - Heroic efforts keep supplies coming in wake of Katrina. PMID- 16268403 TI - Effects of pH on the microhardness of resin-based restorative materials. AB - This study determined the effect of pH on the microhardness of commonly used resin-based restorative materials which included a resin composite (Esthet-X, Dentsply), a new generation compomer (Dyract Extra, Dentsply) and a giomer (Beautifil, Shofu). Fifty-four specimens (3-mm wide x 3-mm long x 2-mm deep) were made for each material. The specimens were divided into six equal groups and conditioned in the following solutions at 37 degrees C for one week: 0.3% citric acid at pH 2.5, sodium hydroxide-buffered citric acid at pH 3, 4, 5, 6 and 7. After conditioning, the specimens were subjected to hardness testing using a digital microhardness tester (load 500gf; dwell time 15 seconds). Data was analyzed using one-way ANOVA and Scheffe's test at a significance level of 0.05. The effects of pH on the microhardness of resin-based restoratives were material dependent. The compomer and giomer materials were more affected by acids of low pH than the composite material that was evaluated. PMID- 16268405 TI - PTCA catheters now a bargain. PMID- 16268406 TI - Interview with Dianne Mandernach, FACHE, Commissioner, Minnesota Department of Health, St. Paul, Minnesota. Interview by Kyle L Grazier. PMID- 16268407 TI - Philanthropy: the last frontier for capital funding. PMID- 16268408 TI - Persuasion: what to say, how to be. PMID- 16268409 TI - Opportunities for administrators to promote disease management. AB - Studies of disease management (DM) have shown that patients who participate in such programs achieve better health status and make fewer emergency room visits. Private and government payers have recently increased their efforts to promote DM initiatives through financial incentives to healthcare providers. This article explores opportunities for administrators of health services organizations (HSO) to promote DM in the current political and economic environment. Our survey of professionals (DM leaders, physicians, and DM nurses) in six DM programs reveals these professionals' assessments of the key players and resources that they deem important to their respective DM programs. They view DM programs as heavily dependent on the support of physicians, nurses, and health plan leaders but relatively less so on the support of HSO administrators- a situation that may suggest opportunities for administrators to take on greater leadership in moving the HSO toward developing DM programs. Survey results also indicate a strong need for the integration of resources such as communication systems, electronic medical records, and DM reporting. Taken collectively, these needs suggest a number of strategies for the administrator to play a larger role in supporting the adoption and effective implementation of DM. In the article, we propose that DM programs can benefit substantially from an administrator who can demonstrate a thorough knowledge of DM-related government and private-payer initiatives and who has the ability to provide leadership to develop and implement viable DM programs. Valued contributions that the administrator should bring to the table include support of standardized DM processes, use of practice guidelines, and provision of pertinent information systems. PMID- 16268410 TI - Managing primary care using patient satisfaction measures. AB - Our study aimed to identify which attributes of a primary healthcare experience have the most impact on patient satisfaction as well as which aspects of each attribute are most significant in patients' response to the services they receive. The three attributes examined in this study were access, staff care, and physician care. Analyses of the aspects of each attribute controlled for age, gender, and race. Data used in this study were obtained through a survey questionnaire with random sampling, resulting in the sample size of 8,465. The psychometric properties of the questionnaire were also examined and showed appropriate reliability and validity. The multiple regression analysis showed that among the three attributes, physician care was most influential, closely followed by staff care, with access having much less influence. Further analyses revealed that specific aspects of each attribute were more influential on patient satisfaction. Within the physician care attribute, patients were found to be rational consumers who were looking for surrogate indicators of correct diagnosis and treatment options among the measures available to them. They were much less likely to be influenced by so-called bedside manner. Within the staff care attribute, willingness and compassionate behaviors of staff and prompt service were most important. Within the access attribute, patients sought caring interaction with appointment personnel. After considering the findings, we discuss possible actions for healthcare managers. PMID- 16268411 TI - Emergency department overcrowding: the impact of resource scarcity on physician job satisfaction. AB - Emergency departments in most developed countries have been experiencing significant overcrowding under a regime of severe resource constraints. Physicians in emergency departments increasingly find themselves toiling in workplaces that are characterized by diminished availability of, limited access to, and decreased stability of critical resources. Severe resource constraints have the potential to greatly weaken the overall job satisfaction of emergency physicians. This article examines the impact of hospital resource constraints on the job satisfaction of a large sample of emergency physicians in Canada. After controlling for workflow and patient characteristics and for various institutional and physician characteristics, institutional resource constraints are found to be major contributors to emergency physician job dissatisfaction. Resource factors that have the greatest impact on job satisfaction include availability of emergency room physicians, access to hospital technology and emergency beds, and stability of financial (investment) resources. PMID- 16268412 TI - Seeding community partnerships in providing medical care that lowers cost of care. PMID- 16268413 TI - Primary antibiotic resistance and effectiveness of Helicobacter pylori triple therapy in ulcero-inflammatory pathologies of the upper digestive tract. AB - OBJECTIVES: To determine firstly, the rates of primary antimicrobial resistance for Helicobacter pylori-associated upper-digestive lesions in relation to the success rate of triple therapy; and secondly, the performance of HpSA stool antigen detection test for control of eradication after treatment. METHODS: Prospective open study of 436 patients who underwent upper-digestive tract endoscopy with biopsies for histological examination and culture between January 1 and July 31, 2002 at a University hospital in Brussels, Belgium. The primary resistance to antibiotics of H. pylori isolates was determined by disc diffusion method. Seventy of 164 infected patients agreed to be included in the treatment study with standard triple therapy with amoxicillin + clarithromycin + omeprazole adjusted on the basis of antibiogram results. Control of eradication was tested by 14C-Urea breath test and H. pylori Stool Antigen test (HpSA test). RESULTS: Primary resistance to clarithromycin and metronidazole was observed in 3% and 31% of the isolates, respectively. No primary resistance to amoxicillin and tetracycline was observed. By intention to treat analysis, H. pylori was eradicated in 56 (80%) patients included in the therapeutic study. Three (4%) patients were lost to follow-up. The rate of eradication failure was 20% (14/70), included 11 cases documented by a positive control test (14C-Urea breath test). In comparison with 14C-Urea breath test, the H. pylori Stool Antigen test showed a sensitivity of 100%, a specificity of 91%, PPV of 69%, and NPV of 100%. CONCLUSION: Standard triple therapy achieved 80% bacterial eradication in this patient population with a low prevalence of H. pylori primary antibiotic resistance. Our data confirm that the H. pylori Stool Antigen test displays a diagnostic performance similar to the breath test for control of eradication. PMID- 16268414 TI - Microsatellite instability in sporadic and inherited colon adenocarcinomas from Greek patients: correlation with several clinicopathological characteristics. AB - BACKGROUND AND AIMS: Microsatellite instability seems to play a significant role in colorectal carcinogenesis, as it is reported to occur in HNPCC patients as well as in a proportion of sporadic cases. The aim of this study was to examine the presence of microsatellite instability in relation to other commonly observed genetic abnormalities and clinicopathological characteristics of sporadic and inherited colorectal cancers. METHODOLOGY: One hundred and three sporadic colorectal adenocarcinomas and 9 adenocarcinomas from HNPCC patients were histologically evaluated. The presence of microsatellite instability was investigated at six loci. K-ras and p53 mutations, p53 LOH, hMLH1 expression and methylation status were examined as well. Statistical analysis was performed to define possible correlations of the observed genetic alterations with the clinicopathological characteristics of the analysed tumors. RESULTS: High-grade microsatellite instability was found in 14% of sporadic adenocarcinomas and in 78% of adenocarcinomas from HNPCC patients. K-ras and p53 mutations were found in 29% and 28% of sporadic adenocarcinomas respectively and in 0% and 22% of the 9 HNPCC cases. A statistically significant correlation was noticed in sporadic tumors between the presence of MSI-H and tumor location at the proximal colon, as well as with the female gender. CONCLUSIONS: Sporadic MSI+ colon adenocarcinomas seem to represent a distinct entity with a unique profile of genetic changes, different from those observed in HNPCC or MSI negative sporadic tumors. PMID- 16268415 TI - Oesophageal intraepithelial and invasive neoplasia of squamous cell type: epidemiology and outcome in Luxembourg, 1980-2001. AB - BACKGROUND AND STUDY AIMS: Oesophageal intraepithelial neoplasia of squamous cell type (INSC) and invasive oesophageal squamous cell carcinoma (IOSCC) are infrequent diseases in Western Europe. The aim of the present study was to collect population-based data of both entities over a 20 year-period and to look for concomitant neoplastic affections in order to define an adequate diagnostic strategy. PATIENTS AND METHODS: The National Morphologic Tumour Registry allowed to review the data of all patients with INSC and IOSCC diagnosed between 1980 and 2001 and to record the time trends in incidence, the oncologic co-morbidity and the outcome of the patients. RESULTS: 29 patients with INSC and 363 cases of IOSCC were identified. The overall age-standardized (world) incidence rate of intraepithelial neoplasia and invasive squamous cell carcinoma were 0.2 and 4.2 per 10(5), respectively, the M/F-ratio for both 3:1. During the study period, the incidence rate of invasive cancer remained stable in males but showed a 3-fold increase in females. There was a 2-fold increase of the intraepithelial neoplasia incidence in the last decade. The precancerous/cancerous-ratio increased slightly over the last 5 years. 31% of the patients with an INSC and 17.6% of those with IOSCC had concomitant precancerous and cancerous lesions especially of head and neck (laryngopharyngeal) or pulmonary origin. The observed 5-year survival rate was 8.8 +/- 3% (95% confidence interval) for IOSCC and 27.6% +/- 17% for INSC. CONCLUSIONS: The incidence of invasive oesophageal squamous cell carcinomas remains stable whereas that of detected intraepithelial squamous cell neoplasias is remarkably low, indicating potential underdiagnosis. Considering the overall low incidence rates, mass screening for oesophageal cancer does not seem reasonable in Luxembourg. Nevertheless, patients at high-risk for oesophageal or head and neck or broncho-pulmonary cancer should be identified and surveilled by endoscopy, possibly with vital staining. PMID- 16268416 TI - Practical use of hepatitis C and B molecular tools: Belgian guidelines. AB - This article discusses the use of virologic assays in the diagnosis and management of hepatitis C virus (HCV) and hepatitis B (HBV) infection. The use of virologic tests has become essential in the management of HCV and HBV infection to diagnose viral infection, guide treatment decisions, and assess the virologic response to antiviral therapy. The continuing development of test systems accompanied by new antiviral drugs and novel therapeutic approaches should lead to an optimization of the treatment of HCV infection. Molecular methods for viral testing have become an integral part of the diagnostic and therapeutic management of infections with hepatitis C virus (HCV) and hepatitis B virus (HBV). PMID- 16268417 TI - The management of patients with mild hepatitis C. AB - Infection with the hepatitis C virus (HCV) represents an important public health problem and is a leading cause of chronic hepatitis, cirrhosis and hepatocellular carcinoma. Chronic hepatitis C is a heterogeneous disease. Many patients have mild disease at presentation but not all of them will develop advanced liver disease. However, the identification of these patients with mild hepatitis C who will show progressive disease is difficult and is based on histological criteria and the assessment of co-factors (age, alcohol intake, steatosis). In addition, serum transaminases that are persistently normal on several occasions during 18 months may point to a more benign course. Patients with mild hepatitis C should not be excluded "a priori" from the possibility of being treated, as treatment with pegylated interferon and ribavirin is safe and effective in this group. Overall, the decision to initiate therapy should be individualized and based on the severity of the disease by liver biopsy, the potential of serious side effects, the probability of response and the motivation of the patient. PMID- 16268418 TI - Stepwise minimization of the immunosuppressive therapy in pediatric liver transplantation. A conceptual approach towards operational tolerance. AB - The evolution of immunosuppression in pediatric liver transplantation has been characterized by a steady reduction of the immunosuppressive load, including removal of anti-lymphocyte antibodies, with the aim to reduce the incidence of EBV-related post-transplant lymphoproliferative disorders. Acute rejection rates were studied retrospectively over two decades of pediatric liver transplantation, according to the successive immunoprophylactic regimens. 318 primary pediatric liver transplant recipients, included between 1984 and 2004 in successive prospective trials, were analyzed, with respect to the impact of the immunosuppressive protocol on acute rejection occurrence. A progressive decrease of rejection incidences was observed, which corresponded to reduced immunosuppressive load and to transplant eras. Such trend might be related to changing approaches towards acute rejection histology and therapy by transplant clinicians, but also to the stepwise minimization of immunosuppressive protocols, putatively enhancing graft acceptance. We hypothesize that the recent population of liver transplant recipients with low immunosuppression might be more suitable for progressive immunosuppression withdrawal trial, with the aim to reach ultimately operational tolerance. PMID- 16268419 TI - Long-term medical complications and quality of life in adult recipients surviving 10 years or more after liver transplantation. AB - BACKGROUND AND STUDY AIMS: Little information is available about long-term results after adult liver transplantation. This study analyses long-term medical complications, changes of immunosuppression, recurrence of primary disease and quality of life 10 years after liver transplantation. MATERIAL AND METHODS: During the period February 1984-April 1994, 324 LT were performed in 282 adults (>15 years). One hundred forty-seven (52%) patients survived more than 10 years. Data regarding health status of 103 patients exclusively followed-up in our institution were analyzed. RESULTS: Actual 1, 5, 10 years survival rates of the 282 recipients were 76.6%, 64.9% and 52% respectively. Forty eight (46.6%) of the 103 studied patients had normal liver tests in their tenth year of the follow-up. Seventy-one (69%) patients were on a CyA, TAC or MMF monotherapy; 31 (30%) patients had CyA levels of less than 100ng/ml. Forty five patients had recurrent allograft disease. Twenty-four (40.6%) of 59 liver biopsy available at 10th year were normal. Thirty five (34%) patients developed chronic renal failure; nine (8.7%) of them had end-stage renal disease. New onset hypertension (>140/100 mmHg) developed in 49 (47.6%) patients; fourteen (13.6%) developed diabetes (glucose blood level > 140 mg/dl) and twenty five (24.2%) patients had serious cardiovascular events. Thirteen (12.6%) patients had a BMI>28 and thirty six (35%) patients had elevated serum cholesterol (>220 mg/dl). Cataract was present in 8 (7.7%) patients. De novo malignancy developed in 23 (22.3%) patients. One patient each developed nasopharyngeal lymphoproliferative disease and myeloma. Quality of life of this patient cohort was excellent as shown by a Karnofsky score of more than 80% in 96.6% of patients. CONCLUSION: The high rate of medical complications and especially of malignant tumours in this long-term follow-up study indicate that further optimization and especially minimization of immunosuppressive therapy as well as development of newer therapies in order to prevent recurrent allograft diseases are the priority for the future development of transplant medicine. PMID- 16268420 TI - Recurrence of autoimmune hepatitis, primary biliary cirrhosis and primary sclerosing cholangitis after liver transplantation. AB - Liver transplantation (LT) is the standard therapeutic approach for the treatment of end-stage acute and chronic autoimmune liver disease as autoimmune hepatitis (AIH), primary biliary cirrhosis (PBC) and primary sclerosing cholangitis (PSC). Results of liver transplantation in these indications are good with a patient survival after LT at 5 years of 85%. However several series have reported a possible recurrence of primary autoimmune liver disease after liver transplantation. Concerning all these three autoimmune liver diseases, recurrence of the disease on the graft may have multiple clinical, biochemical, histological and radiological expression influenced by different factors as the diagnostic methods used, the degree of immunosuppression and the genetic background of the recipient. We would like with this overview to describe the different pattern of recurrence of these autoimmune liver disease, their potential influence on the liver graft and their therapeutic management. PMID- 16268421 TI - Recurrent allograft disease: viral hepatitis. AB - Viral hepatitis is the leading indication for liver transplantation (LT) in the majority of transplant centers. Post-transplantation outcome in these patients largely depends on the prevention of allograft reinfection. In contrast to hepatitis B where excellent results have been achieved following the implementation of effective measures to prevent HBV, recurrent hepatitis C is an increasing problem facing liver transplant hepatologists and surgeons. HBV recurrence is effectively contained by the use of hepatitis B inmunoglobulins with antivirals. Unfortunately, no effective prophylactic therapy is available for hepatitis C so that recurrent hepatitis C occurs almost invariably. Progression to severe allograft fibrosis is often rapid. Current antivirals, including peg-interferons, are limited by substantial toxicities that compromise their efficacy. Hence, it is not surprising that although some improvements have been made in the treatment of recurrent hepatitis C, a substantial proportion of HCV-infected patients develop recurrent allograft end-stage liver disease leading to a decrease in graft survival, an increase in the need for re-transplantation, and ultimately, a decrease in patient survival. PMID- 16268422 TI - Noncompliance with immunosuppressive regimen in organ transplantation: is it worth worrying about? PMID- 16268423 TI - Transplantation okay--psyche okay? Reflections on psychosomatics in the field of organ transplantation. AB - Transplantation puts a large burden on patients' psyche, before and after the operation. Psychosomatic care implicates helping patients to take a firm decision in favour of a new organ, of a new life. Incorporation of the graft, efficient doctor-patient-relations, pregnancy and sexuality, everything is possible but crucial to many patients. Psychosomatic knowledge and specified consulting help them and their families and even the doctors and nurses to cope with overwhelming emotions, fear and a lifelong danger of losing the organ. Transplantation means crossing borders, going into unknown psychic regions. And the recent rapid development of living liver transplantation does not facilitate things. PMID- 16268424 TI - Prospects of the use of hepatic cells for extracorporeal liver support. AB - Hybrid extracorporeal liver support is an option to assist liver transplantation therapy. An overview on liver cell bioreactors is given and our own development is described. Furthermore, the prospects of the utilization of human liver cells from discarded transplantation organs due to steatosis, cirrhosis or traumatic injury, and liver progenitor cells are discussed. Our Modular Extracorporeal Liver Support (MELS) concept proposes an integrative aproach for the treatment of hepatic failure with appropriate extracorporeal therapy units, tailored to suit the actual clinical needs of each patient. The CellModule is a specific bioreactor (charged actually with primary human liver cells, harvested from human donor livers found to be unsuitable for transplantation). The DetoxModule enables albumin-dialysis for the removal of albumin-bound toxins, reducing the biochemical burden of the liver cells, and replacing the bile excretion of hepatocytes in the bioreactor. A Dialysis Module for continuous veno-venous hemofiltration can be added to the system if required in hepato-renal syndrome. PMID- 16268425 TI - Extension of the adult hepatic allograft pool using split liver transplantation. AB - BACKGROUND: The ever increasing number of, especially, adults waiting for a liver transplantation necessitates to develop techniques allowing to extend the available donor liver pool. MATERIALS AND METHODS: Between November 1988 and December 2004, 37 (6.6%) of 559 adults underwent split liver transplantation at Saint-Luc Hospitals. There were 36 were right and one left split procedures; 27 split grafts were obtained ex-situ and 10 in-situ. Results of these series are analysed and compared to literature data of split liver transplantation. RESULTS: Three and 12 months patient survival rates were 89.2% and 78.4% respectively. Five years actuarial patient survival was 75.7%. Early (< 3 months) and late (> 3 months) mortality rates were 10.8% (4 pat.) and 21.6% respectively. Early mortality was significantly higher in case of urgent split liver transplantation (3/5 patients vs. 2/32 elective patients--p 0.001). At present 25 patients are alive, with a mean Karnofsky score of 90%. Three and 12 months graft survival rates were 91.7% and 87.1% respectively. Three and one grafts were lost due to primary and early graft non-function. In-situ split grafts had shorter mean warm, cold, total ischemia and operating times as well as less need for blood transfusion; all these differences were however not statistically significant. Surgical complications occurred in 19 (51%) patients. All but one complication occurred early (< 3 months). There were sixteen biliary complications in 13 (35.1%) patients: 9 anastomotic stenoses, 3 anastomotic and 4 transection margin leakages. Six vascular complications occurred in 6 (15.2%) patients: three arterial and 3 portal vein thromboses. Seven (18.9%) patients had a postoperative bleeding. CONCLUSIONS: Graft and patient survival rates of split liver transplantation can be compared to those of classic liver transplantation. However the care of these patients is demanding due to the high number of technical complications. Results of split liver transplantation must be further improved in order to foster it's more widespread use necessary to overcome the actual shortage of liver allografts. PMID- 16268426 TI - Regression of Sweet's syndrome associated with Crohn's disease after anti-Tumour Necrosis Factor therapy. AB - The association of inflammatory bowel disease and acute febrile neutrophilic dermatitis (Sweet's syndrome) has infrequently been reported in the literature. We describe the case of a 41-year-old Caucasian woman with ileo- anal Crohn's disease who presented simultaneously an erythema nodosum and a Sweet's syndrome. A dramatic regression of the cutaneous lesions was observed after infliximab treatment, indicating that this therapy might be useful for both Crohn's disease and Sweet's syndrome. PMID- 16268427 TI - Very high elevation of CA19-9 level in a patient with steatosis. PMID- 16268428 TI - A succesfully managed inflammatory pseudotumour of liver without surgery: report of a case. AB - The inflammatory pseudotumor (IPT) of the liver is an extremely rare focal lesion of the parenchyma with unknown aetiology. It has the appearance of a malignant tumor but has a benign histology and clinical course. Herein, we report a case of IPT in a 55 year-old woman. She had complained of right upper quadrant pain and subfebrile fever. Imaging studies revealed a heterogeneous, solitary mass in right lobe of liver which was 90 x 81 mm in size. Histopathological examination of ultrasonography-guided true-cut biopsy demonstrated IPT. After the initiation of the antibiotic and nonsteroidal anti-inflammatory drugs (NSAID), the size of hepatic pseudotumor decreased (20 x 20 mm) and then completely disappeared. To rule out a malignancy and to reach the diagnosis of IPT, biopsy from the mass is mandatory. After the diagnosis, IPT gave an excellent response to short course of antibiotic and NSAID therapy. To prevent unnecessary resective liver surgery, IPT should be kept in mind in the differential diagnosis of liver mass which can be successfully managed medically. PMID- 16268429 TI - Subacute cholestatic hepatitis likely related to the use of senna for chronic constipation. AB - We report a case of senna-induced cholestatic hepatitis which was not diagnosed at presentation. A 77 year old male was referred with abdominal pain, jaundice and elevated transaminase levels. A diagnosis of extrahepatic cholestasis was first suspected, due to the observation of a duodenal diverticulum and dilated proximal choledocus. However, the sphincterotomy did not improve cholestasis. At further evaluation, HBsAg was positive but serological work up was compatible with a healthy-carrier status. Further interrogation of the patient revealed a history of chronic senna intake to treat a chronic constipation. Liver biopsy showed bridging hepatocellular necrosis as well as canalicular cholestasis. Drug withdrawal resulted in a slow and progressive reduction in bilirubin levels and liver enzymes. In this case senna was likely the cause of a subacute cholestatic hepatitis exemplifying again the potential role of herbal related liver injury. PMID- 16268431 TI - Fatal acute sodium phosphate enemas intoxication. AB - We describe a patient who died as a result of severe hypocalcaemia and hyperphosphatemia after treatment with a sodium-phosphate enema. Physicians should be aware of the risk when using these enemas, even in normal doses, especially in elderly patients without signs of renal failure, as in our patient. PMID- 16268430 TI - Upper gastrointestinal complaints as a consequence of thoracic spinal tumor. AB - A rare case of a 43-year-old male with chronic epigastric complaints and atypical diffuse osseous pain for two years, which were finally found to be caused by a benign thoracic spinal tumor (lipoma) and was successfully treated by neurosurgical resection, is presented. At three years follow-up postoperatively he remains completely asymptomatic. This report discusses the case and the potential pathophysiology of the patient's symptoms. PMID- 16268432 TI - Rare complication of interferon alpha therapy: retinal vein thrombosis. PMID- 16268433 TI - Committee for the protection of persons. AB - The transposition into French law of Directive 2001/20/CE, which relates to good clinical practice in the conduction of trials on drugs for human use, has required the modification of certain provisions that concern the protection of persons participating in biomedical research, in particular those provisions concerning the conditions for the authorisation of biomedical research. Declaration to the competent authorities now comes prior to authorisation and, henceforth, the prior opinion of a Committee for the Protection of Persons (CPP) must be expressly favourable in order for a trial to be undertaken. Proposals are put forward by this Round Table in order to promote the stability and professionalism of the CPPs. PMID- 16268435 TI - The human biological resource centres network. A key infrastructure for biomedical research in Europe. AB - Continuing advances in life sciences and medical research are the result of the remarkable achievements of cell and molecular biology. In the post-sequencing era, the quality of the huge amount of data continuously generated by biotechnology, i.e. genomics, proteomics and high-throughput screening, depends on the quality assurance and the traceability of the original biological materials, including the annotations linked to these materials. Thus, biological resource centres are key infrastructures supporting biotechnology, bioprocessing and the development of new approaches in the prevention, diagnosis and treatment of diseases. PMID- 16268434 TI - The respective roles of controlled clinical trials and cohort monitoring studies in the pre- and postmarketing assessment of drugs. AB - The respective roles of controlled clinical trials and observational studies (cohort or case-control studies) in evaluating the efficacy, safety and usefulness of a drug were analysed. A randomised, controlled, double-blind study is the best method of estimating the efficacy of a treatment. It provides the least biased and most robust estimate of the causal relationship. In certain situations and on the basis of certain criteria, observational studies can have a proof-of-efficacy value. Randomised, controlled, pre- and postmarketing authorisation (MA) clinical studies identify the rarer adverse effects and compare them with those resulting from the reference treatment. Before the MA, the pooled safety data from different controlled trials can provide an estimation of relatively frequent adverse events and subjects at risk. However, an observational study is the most appropriate method of evaluating the safety of a drug in the currently used conditions. By definition, a drug influences the health of a population if it directly or indirectly improves its health. A drug would have a major role in public health if it reduced mortality or morbidity related to a particular disease or if it improved the quality of life of patients with this disease. Prior to marketing a product, modelling is the approach of choice to quantify the expected effect. Pragmatic, postmarketing trials and observational studies are the reference methods used to define the population affected, the efficacy and safety of the drug in a real situation and its usefulness for public health. In conclusion, randomised clinical trials remain the reference approach for evaluating efficacy, while observational studies have a confirmatory value. Observational studies are the most appropriate way of evaluating safety in the currently used conditions, as the clinical trial has limited indications. In the interests of public health, modelling is the pre marketing approach of choice, while pragmatic trials and observational studies are the postmarketing reference approaches. PMID- 16268436 TI - Methodology for the evaluation and measurement of therapeutic progress. AB - When a candidate drug is likely to become available to prescribers and healthcare policy makers, evaluation of therapeutic progress moves forward in two stages. First, the level of expected therapeutic progress must be established. This first stage requires the determination of therapeutic needs and the comparison of these against the results of the clinical studies that will form the basis of the marketing authorisation of the drug. This determination helps anticipate the therapeutic progress that is attributable to the approved use of the new drug. The second stage of the process, the evaluation of the actual therapeutic progress, involves therapeutic drug monitoring and bases itself on observation. Since such observational data are intended to challenge the initial hypotheses and uncertainties (in terms of benefits and risks), goals and methods must be laid out before the drug becomes available to the general public. PMID- 16268437 TI - Appropriate medication use in individuals aged 80 or more years. AB - Over the last few years, there has been a rapid and steadily expanding demographic explosion in the elderly population in France. When the most dynamic segment of this growth is considered, i.e. patients aged 80 years or more (who, moreover, usually have several diseases), under- or over-prescribing and/or inappropriate prescribing are identified. Three short- and medium-term recommendations are proposed: to quickly get expert consensus on the theme of prescribing for the aged patient; to integrate systematic drug re-evaluation into annual consultations for people aged 80 years or more; and to better coordinate actions and communication between institutions, health professionals and the pharmaceutical industry. PMID- 16268438 TI - Modelling and clinical trials in paediatrics. AB - Clinical trials are more difficult to conduct in children, but they are even more necessary than in adults their scarcity is an ethical scandal. Mathematical models can be built that can describe both the disease process and the mechanism of action of drugs. These models can then be used to simulate the outcome of clinical trials. Inspection of the simulated results then facilitates optimisation of the trial design and proposed methods of analysis. Validation is a crucial issue for the good practice of modelling and simulation. The participants of Round Table No. 6 recommend: (i) that modelling be systematically employed; (ii) that all the required professional personnel be involved, at all phases; (iii) that all data needed are made accessible; (iv) that clinicians be trained; (v) that specialists develop training tool kits; and (vi) that universities provide appropriate training. PMID- 16268439 TI - [Medication errors resulting from drug preparation and administration]. AB - BACKGROUND: The risks associated with drug use are not confined to adverse reactions. Failures can occur in the process of drug prescribing, dispensing and administration. Such preventable events are termed 'medication errors'. Errors in preparation and administration, the last step in the medication process, constitute a good indicator of the quality of the medication process, and are irredeemable. METHODS: A protocol for measuring errors in the preparation and administration of medication has been developed and used in an internal medicine department at the University Hospital of Dijon. RESULTS: This protocol has enabled several different rates of medication errors to be determined depending on the calculation approach used: 15.1% with respect to interventions by nurses, 41% relating to the total number of opportunities for error, and 8.8% with respect to a methodological problem analysed in the study. The potential clinical significance, incidence and causes of errors during the medication process were also analysed. DISCUSSION AND CONCLUSION: Medical errors are not detected in our health system and are thus not preventable. Their consequences are incompatible with a well organised treatment process. The rate of medication errors is therefore a good indicator of the quality of the medication system in a hospital that is following the current steps for risk reduction and accreditation. PMID- 16268440 TI - [Prescribing drugs by international non-proprietary name: the perception of health and non-health professionals]. AB - AIM: The aim of this study was to evaluate the perceptions of health professionals and non-health professionals with regard to prescribing drugs by their international non-proprietary name (INN) in the Midi-Pyrenees area, France. METHODS: We developed a score to assess the perception of the four criteria that make up therapeutic progress: efficacy, safety, convenience and cost for the National Health Insurance. Changes in perception under these criteria were scored between 0 and 10 (0 for no change and 10 for maximal change). The questionnaire was answered by 142 general practitioners, 161 pharmacists and 132 healthy subjects (public). RESULTS: The median value (first quartile to third quartile) for the perceived change in efficacy was 0 (0-3) for physicians and pharmacists, and 0 (0-0) for the public. The median value for the perceived change in safety was 0 (0-3) for practitioners and pharmacists, and 0 (0-5) for the public. The median value for the perceived change in convenience was 0 (0-6) for practitioners, 5 (0-6) for pharmacists and 0 (0-0) for the public. The median value for the perceived change in cost was 6 (3-8) for practitioners, 8 (5-10) for pharmacists and 10 (6-10) for the public. CONCLUSION: This study shows that there is generally favourable acceptance of prescribing by INN by those in the health area. However, general practitioners seem to be more reluctant to accept this than pharmacists or the general public. PMID- 16268441 TI - [Evaluation of self-medication by scuba divers]. AB - AIM: The aim of the study was to evaluate drug use amongst French scuba divers. METHODS: An anonymous questionnaire was distributed to 220 people, out of which 106 responded. RESULTS: Fifteen percent of respondents were receiving long-term medication whereas 60.33% of respondents reported taking medication only occasionally when diving. Thirty-seven percent of respondents were taking prescribed medicines. Drugs consumed occasionally included analgesics (43.5%), corticosteroids (14.5%) and antiemetics. A detailed study of aspirin and steroid consumption revealed their misuse for performance-enhancing purposes, the level of risk varying with the type of diving activity. The influence of drug use on personal performance was recognised by 72.6% of divers, but only 59.4% considered that they were putting themselves at risk by self-medicating. PMID- 16268442 TI - [Clinical trials using medicinal plants: bibliographical review and methodological analysis]. AB - BACKGROUND: The importance of traditional medicine, one of the fundamentals of the cultural heritage of African, Asian and South American peoples, is evident in that such medicine is practised by more than 80% of these populations. METHODS: To analyse the methodology of clinical trials using medicinal plants, we reviewed articles published on this topic between 1980 and 2000. RESULTS: Forty-eight clinical trials were identified. Most were carried out in developed countries. Standard methodological principles were applied in almost all the trials: randomisation (85.4%), comparison (87.5%) versus placebo (95.2%), and blinded design (81.3%). The duration of the studies was short. Sample sizes were generally small, ranging from 30 to 99 subjects; statistical tests were used in 90% of the trials. Adverse effects were infrequently collected. CONCLUSION: Most clinical trials included in this survey were conducted in accordance with WHO guidelines. Respect for methodological principles and the implementation of a legislative framework are important in obtaining credibility and international recognition of the traditional pharmacopoeia. PMID- 16268443 TI - Trimetazidine induces parkinsonism, gait disorders and tremor. AB - OBJECTIVE: To study the adverse effects of trimetazidine on motor functions. DESIGN: A retrospective study was carried out using electronic records to identify all patients seen between January 1990 and August 2003. SETTING: A neurological out-patient clinic. PARTICIPANTS: Of the 10 258 patients who attended the clinic, 130 received trimetazidine. Treatment with this drug was discontinued in 128 patients. Of the 130 patients treated with trimetazidine, 29 also had other drugs capable of inducing parkinsonism withdrawn from their treatment. MAIN OUTCOME MEASURES: Identification of an improvement in motor function after drug withdrawal. RESULTS: In 56 of the 130 patients who were treated with trimetazidine (43%), an adverse effect on motor function was detected that had been induced or aggravated by one of the withdrawn drugs. Indeed, drug-induced parkinsonism was detected in 20 of these patients. Of these, ten were being treated with trimetazidine only, while the remaining ten were simultaneously receiving other drugs potentially capable of inducing parkinsonism. Treatment with trimetazidine worsened previously diagnosed Parkinson's disease in 12 patients, and gait disorders coupled with disequilibrium was observed in 15 patients. Trimetazidine induced tremor in nine patients. CONCLUSION: Trimetazidine can induce parkinsonism, gait disorder and tremor. These adverse effects have not been previously described for this drug. PMID- 16268445 TI - [Osteoporosis: a controlled epidemic?]. PMID- 16268444 TI - [Severe long-term furosemide (Lasilix) abuse: dependence syndrome or not?]. PMID- 16268446 TI - [Should we detect osteoporosis, and how?]. AB - Osteoporotic vertebral and hip fractures, which are very frequent among elderly, are associated to an excess of mortality, as well as to an important morbidity. In Switzerland, annual costs of in-patients' treatments of osteoporosis are beyond CHF 700 millions. As several effective treatments against osteoporosis are available, a systematic screening is justified. The methodical search for risk factors of fracture and the recognized medical indications for bone densitometry are of prime necessity. The appreciation of clinical symptoms associated to prevalent vertebral fractures (wall-occiput distance, ribs-pelvis distance) is an essential component to patients' approach. Thanks to technological progress in the field of DXA, which allows a visualisation of the spine from L4 to T4, this screening should become easier. PMID- 16268447 TI - [Is vertebral fracture diagnosed?]. AB - Vertebral fractures are the most common osteoporotic fractures. Higher morbidity, higher mortality and higher risk of future osteoporotic fractures are associated to them. However, vertebral fractures are largely under-recognized and therefore, a large part of osteoporotic patients remains untreated. An education program which emphasises the importance of vertebral fracture and proposes a detection method applicable in daily practice should improve the diagnosis and the management of osteoporotic patients. PMID- 16268448 TI - [Osteoporosis and anti-androgenic therapy in case of prostate cancer]. AB - Anti-androgenic therapy for prostate cancer is associated with an increased risk of osteoporosis and of fracture. A fracture occurence has negative influence on survival. An antiresorptive agent like zoledronate, prevents bone loss consecutive to anti-androgenic therapy. PMID- 16268449 TI - [Bisphosphonates and combined treatments osteoporosis]. AB - Bisphosphonates are inhibitors of bone resorption, which reduce vertebral and non vertebral fracture risk. Their efficacity has been shown in controlled trials up to 5 years, and their safety up to ten years. There is no proven benefit of combining two bone resorption inhibitors. PMID- 16268450 TI - [Parathyroid hormone for osteroporsis: why and who to treat?]. AB - Osteoporosis treatment with anti-catabolic agents, such as bisphosphonates and SERMS, inhibits bone remodeling, resulting in increased bone mineral density and preservation of bone micro-architecture. On another side, daily subcutaneous injections of parathyroid hormone (and PTH(1-34), teriparatide) stimulate osteoblastic functions and bone remodeling, resulting in new bone formation, increased bone mineral density and improved microarchitecture, thereby reducing fracture risk. However, the effects on PTH on cortical bone structure and its use in the context of anticatabolic treatments for osteoporosis remain complex. PMID- 16268451 TI - [Adherence to treatment in osteoporosis influence on efficiency]. AB - Low adherence to therapies has been repeatedly described as a major determinant of poor clinical outcomes in chronic disorders. Bisphosphonates, the most widely prescribed drugs in this field, have been linked to a 12-month persistence lower than 40%. The situation is improved when using the weekly formulation compared to the daily intake of the drug. Low compliance results in lesser increase in bone mineral density and decreased anti-fracture efficacy. New medications, currently developed for the management of osteoporosis, will be user-friendly, allowing to an improvement of compliance and persistence. PMID- 16268452 TI - [Cancer of the esophagus: news]. PMID- 16268453 TI - [Avian influenza, the new menace that hangs over our sky]. PMID- 16268454 TI - [Preimplantation diagnosis--bioethical issues surrounding a current patient]. PMID- 16268455 TI - [Medicine as a test of human credulity]. PMID- 16268456 TI - [Conduct disorder in children and adolescents: what is the role of medicine?]. PMID- 16268457 TI - [Behind the veil]. PMID- 16268458 TI - Tissue engineering of heart valves--hype or hope? PMID- 16268459 TI - Carboxypeptidase U (TAFIa): a metallocarboxypeptidase with a distinct role in haemostasis and a possible risk factor for thrombotic disease. AB - Since the discovery of Carboxypeptidase U (CPU) in 1988, considerable information has been gathered about its biochemistry and function in physiological and pathophysiological circumstances. A variety of tools such as assays to measure proCPU and CPU, antibodies raised against (pro)CPU, selective CPU inhibitors and knock-out mice have been developed and are currently being used to explore the role of this metallocarboxypeptidase in different in vivo and in vitro settings. The knowledge that proCPU can be activated by thrombin and plasmin, enzymes with a key function in coagulation and fibrinolysis, and the ability of CPU to remove C-terminal lysine residues has led to the hypothesis that the proCPU/CPU pathway plays a role in the balance between coagulation and fibrinolysis. The maintenance of the equilibrium between coagulation and fibrinolysis is crucial for normal haemostasis and disturbance of this delicate balance can lead either to bleeding tendency or thrombosis. This review provides an update on several aspects of CPU known at the moment, including an extensive overview on the clinical studies performed up till now. PMID- 16268460 TI - The potential role of platelet microparticles in atherosclerosis. AB - The release of microvesicles ('platelet microparticles', PMPs) by activated platelets has been shown to be an integral part of the thrombotic process. PMPs are believed to mediate many biological processes as they possess various platelet membrane proteins and bioactive lipids. Of note, there is a growing body of evidence that PMPs are involved in all stages in the pathobiology of atherosclerosis. In addition to their role in thrombosis, PMPs may also have a pro-inflammatory effect, which promotes the development of atherosclerosis. Also, bioactive lipids in PMPs have been shown to have important effects on angiogenesis. This review summarises the various studies on the possible role of PMPs in the progression of atherosclerosis. PMID- 16268461 TI - Generation of genetically-altered mice producing very low levels of coagulation factorVII. AB - It has been shown earlier that mice with a total targeted deletion of the factorVII gene (FVII(-/-)) die perinatally, thereby precluding study of adult animals with this total deficiency. Consequently, mice producing very low levels of FVII were developed by targeted replacement of the wild-type (WT) murine FYII gene with its corresponding cDNA, under control of the tetracycline transactivator (tTA) promoter. When backcrossed into the C57BI/6 strain, unchallenged mice containing two replaced FVII(tTA) alleles (FVII(tTA/tTA) produce approximately 0.7% of WT FVII levels, but yet live to adulthood despite displaying severely downregulated overall thrombin production and spontaneously developing cardiac fibrosis at a young adult age. This genetically-altered mouse line provides an excellent animal model to study consequences of a severe FVII deficiency in unchallenged mice and in mice subjected to a variety of experimental challenges. PMID- 16268462 TI - A murine model of deep vein thrombosis: characterization and validation in transgenic mice. AB - Deep vein thrombosis (DVT) occurs with high prevalence in association with a number of risk factors, including major surgery, trauma, obesity, bed rest (> 5 days), cancer, a previous history of DVT, and several predisposing prothrombotic mutations. A novel murine model of DVT was developed for applications to preclinical studies of transgenically constructed prothrombotic lines and evaluation of new antithrombotic therapies.A transient direct-current electrical injury was induced in the common femoral vein of adult C57BI/6 mice. A non occlusive thrombus grew, peaking in size at 30 min, and regressing by 60 min, as revealed by histomorphometric volume reconstruction of the clot. Pre heparinization greatly reduced clot formation at 10, 30, and 60 min (p < 0.01 versus non-heparinized). Homozygous FactorV Leiden mice (analogous to the clinical FactorV Leiden prothrombotic mutation) on a C57Bl/6 background had clot volumes more than twice those of wild-types at 30 min (0.121 +/- 0.018 mm3 vs. 0.052 +/- 0.008 mm3, respectively; p < 0.01). Scanning electron microscopy revealed a clot surface dominated by fibrin strands, in contrast to arterial thrombi which showed a platelet-dominated structure. This new model of DVT presents a quantifiable approach for evaluating thrombosis-related murine transgenic lines and for comparatively evaluating new pharmacologic approaches for prevention of DVT. PMID- 16268463 TI - Postprandial elevation of tissue factor antigen in the blood of healthy adults. AB - Atherosclerosis is a dynamic disease involving lipid metabolism, inflammation and thrombosis. A key factor in thrombosis is tissue factor, a small transmembrane glycoprotein. Tissue factor binds FactorVIIa, and this complex converts Factor X to Factor Xa, leading to thrombin generation and fibrin formation. Inhibition of this pathway is by tissue factor pathway inhibitor (TFPI). Tissue factor is found sequestered within atherosclerotic plaques, and plaque rupture allows tissue factor exposure to the circulation, leading to formation of a thrombus. Tissue factor is also associated with membrane microparticles in the circulation, most likely released from monocytes activated by an inflammatory event. We hypothesize that consumption of a typical western diet that is moderate in fat content leads to elevated levels of circulating tissue factor that may act as a marker of a prothrombotic state. Healthy volunteers, aged 18-55, consumed a moderate (40%) fat meal, with blood taken before and 3.5 and 6 h after the meal. Plasma was isolated and assayed for plasma triglycerides, tissue factor, thrombin antithrombin (TAT) complexes, TFPI and TNFalpha. The levels of circulating tissue factor increased 56% (from 78 pg/ml to 120 pg/ml) 3.5 h after the meal. Levels decreased, but had not returned to baseline 6 h postprandially. No significant differences in TAT, TFPI and TNFa levels were observed postprandially. These results demonstrate increased tissue factor levels in individuals who consumed a moderate fat diet. This suggests that the typical western diet may play a larger role in cardiovascular disease than merely altering lipid profiles. PMID- 16268464 TI - Association of MTRRA66G polymorphism (but not of MTHFR C677T and A1298C, MTRA2756G, TCN C776G) with homocysteine and coronary artery disease in the French population. AB - Methylenetetrahydrofolate reductase polymorphism (MTHFR C677T) is an established determinant of homocysteine plasma level (t-Hcys) while its association with coronary artery disease (CAD) seems to be more limited. In contrast, the association of the substitutions A2756G of methionine synthase (MTR), A66G of methionine synthase reductase (MTRR) and C776G of transcobalamin (TCN) to both t Hcys and CAD needs to be evaluated further. The objective was to evaluate the association of these polymorphisms with t-Hcys and CAD in a French population. We investigated the individual and combined effects of these polymorphisms and of vitamin B12 and folates with t-Hcys in 530 CAD patients and 248 matched healthy controls. t-Hcys was higher in the CAD group than in controls (11.8 vs 10.4 microM, P < 0.0001) and in carriers of MTRRAA and MTHFR 677TT than in those carrying the most frequent allele of both polymorphisms (13.8 vs 11.4 microM, P = 0.0102 and 12.5 vs 11.0 mM, P = 0.0065 respectively). The frequency of MTRR A allele was higher in CAD patients than in controls (0.48 [95% CI: 0.44-0.52] vs 0.38 [95% CI: 0.32-0.44], P = 0.0081) while no difference was observed for MTHFR 677T frequency. In multivariate analysis, t-Hcys > median and MTRRAA genotype were two significant independent predictors of CAD with respective odds ratios of 3.1 (95 % CI: 1.8-5.1, P < 0.0001) and 4.5 (95% CI: 1.5-13.1, P = 0.0051). In conclusion, in contrast to North Europe studies, MTRRAA genotype is a genetic determinant of moderate hyperhomocysteinemia associated with CAD in a French population without vitamin fortification. PMID- 16268465 TI - Significant association with location of central venous line placement and risk of venous thrombosis in children. AB - Venous thromboembolic events (VTE) in children are frequently associated with central venous lines (CVL). Identifying risk factors related to CVL management could potentially minimize CVL-related thrombotic complications. The objectives of the study were to assess whether CVL location, type, size, and duration of placement are associated with the incidence of VTE in children. The study was a prospective, multicentre cohort study in a general pediatric population requiring CVL. Data on CVL characteristics were documented prospectively using standardized case report forms. Outcome assessments were by i) clinical monitoring for symptomatic VTE which were confirmed by appropriate objective test, or ii) screening by venography at study exit. Among 158 children, 21 (13%) hadVTE. The incidence of VTE was increased with femoral CVL (32%) and subclavian CVL (27%) compared to brachial CVL (12%) and jugular CVL (8%; p = 0.01). The incidence of VTE was independent of CVL type (peripherally inserted central catheters, untunneled CVL, tunneled exteriorized CVL, subcutaneous ports; p = 0.90), and CVL size (CVL diameter, p = 0.42; number of CVL lumen, p = 0.58). The incidence of VTE did not increase with duration of CVL placement: 0-5 days (17% VTE), 6-20 days (19%), 21-35 days (10%), and 36-50 days (11%, p = 0.68). The incidence of CVL-relatedVTE may be reduced by preferred placement of CVL in brachial or jugular veins. The choice of CVL type and size does not significantly influence the risk of VTE. Short-term CVL are associated with a similar risk of VTE as longer-term CVL. PMID- 16268466 TI - Prognostic factors for recurrence of venous thromboembolism (VTE) or bleeding during long-term secondary prevention of VTE with ximelagatran. AB - The oral direct thrombin inhibitor ximelagatran (24 mg twice daily) has been shown to significantly reduce the incidence of recurrent venous thromboembolism (VTE) vs. placebo over 18 months, with no significant influence on bleeding (THRIVE III). The influence of potential prognostic factors on the risk of recurrent VTE or major and/or minor bleeding and their impact on ximelagatran treatment was evaluated in the THRIVE III study population. The effect of sex, age, body weight, renal function, malignancy, type of initial VTE event, and history of previous VTE events was investigated in the intention-to-treat population using Cox proportionate hazard modelling. Ximelagatran was administered to 612 patients and placebo to 611 patients. Within the placebo group, risk of recurrent VTE was higher among men than women (hazard ratio [HR]: 2.50,95% confidence interval [CI] 1.49,4.17), and in patients with one or more than one previous VTE event (HR: 1.73,95% CI 1.00, 2.99). There was a higher risk of bleeding among women than men in both the ximelagatran (HR: 1.49, 95% CI 1.06, 2.09) and placebo (HR: 1.48, 95% CI 1.01, 2.15) groups, and in placebo-treated patients with an initial pulmonary embolism (HR: 1.53, 95% CI 1.06,2.23) compared to those with initial deep vein thrombosis. There were no significant interactions between treatment effect and any of the potential prognostic factors. In conclusion, the superior efficacy of ximelagatran vs. placebo was maintained in all subgroups. Long-term use of oral ximelagatran, without coagulation monitoring or dose adjustment, should be feasible and well tolerated in a wide cross-section of patients for the secondary prevention of VTE. PMID- 16268467 TI - Bridging anticoagulation with low-molecular-weight heparin after interruption of warfarin therapy is associated with a residual anticoagulant effect prior to surgery. AB - Bridging anticoagulation with low-molecular-weight heparin (LMWH) is common in patients who require temporary interruption of warfarin before surgery or a procedure, but whether such patients have a residual anticoagulant effect just before a procedure is not known. Consecutive patients who received bridging anticoagulation with LMWH had anti-Xa levels measured just before a procedure. The proportion of patients with a residual anticoagulant effect, defined as an anti-Xa level > or = 0.10 IU/ml, was determined. Multivariable regression analysis identified predictors of a residual anticoagulant effect, expressed as an odds ratio (OR) and corresponding 95% confidence interval (CI). A pre procedure residual anticoagulant effect was detected in 12 of 73 (16%) patients overall, in 11 of 37 (30%) patients who received therapeutic-dose LMWH, and in 1 of 36 patients (3%) who received low-dose LMWH. Receiving therapeutic-dose LMWH (OR = 118.8; 95% CI: 5.8, 999.9), and increasing age (OR = 4.0; 95% CI: 1.3, 12.5) were predictors of a residual pre-procedure anticoagulant effect. In patients who require bridging anticoagulation with LMWH, a residual anticoagulant effect from LMWH is detected in 1 of 6 patients, and receiving therapeutic-dose LMWH is the strongest predictor of such an effect. PMID- 16268468 TI - Rates of proximal deep vein thrombosis as assessed by compression ultrasonography in patients receiving prolonged thromboprophylaxis with low molecular weight heparin after major orthopedic surgery. AB - Patients undergoing major orthopedic surgery are at an increased risk of thromboembolism even after the acute postoperative phase. Therefore, prolonged thromboprophylaxis is currently recommended and widely used. The length of hospital stay after major orthopedic surgery is steadily decreasing and most patients are transferred to specialized rehabilitation centers in the early postoperative phase. Consequently, thromboprophylaxis is usually given during the rehabilitation period. Previously there have been no systematic studies of how many patients with asymptomatic thrombosis are transferred to a rehabilitation center or how many patients develop deep vein thrombosis (DVT) during rehabilitation. In the present study, 238 patients who had undergone major orthopedic surgery were examined by compression ultrasonography on day 1 or 2 and on day 20 of their rehabilitation phase in order to detect asymptomatic proximal DVT. Sonograms were read centrally with blinding to the clinical course of the patients. All patients received standard thromboprophylaxis during the acute postoperative phase. Prolonged thromboprophylaxis with enoxaparin was given during the rehabilitation phase. The mean duration of thromboprophylaxis was 36.8 days. At admission to the rehabilitation center, proximal DVT was identified in 16 patients (6.7%). New proximal DVT at discharge from the rehabilitation center was identified in 3.2% of patients following prolonged thromboprophylaxis with enoxaparin. In conclusion, a considerable percentage of patients with asymptomatic proximal DVT are transferred to rehabilitation centers following major orthopedic surgery. Using prolonged thromboprophylaxis, new proximal DVTs will still develop during rehabilitation, but at a low rate. PMID- 16268470 TI - An association between oxygen desaturation and D-dimer in patients with obstructive sleep apnea syndrome. AB - D-dimer, a degradation product of fibrin, is being increasingly used as a marker and prognostic factor in various thrombotic diseases. Previous reports have shown that obstructive sleep apnea is associated with platelet activation and hypercoagulability. The aim of the study was to assess the potential role of the plasma D-dimer test in patients with obstructive sleep apnea. We designed a prospective group comparison study in a tertiary-care, university-affiliated medical center. One hundred and three patients of mean age 57 years (range 50-76 years) with symptoms suggestive of obstructive sleep apnea were included. Polysomnography was performed in all cases, and blood was collected for plasma D dimer measurement by MiniQuant turbidmetric assay. The demographic and polysomnograph data were compared between patients with normal and high (> 250 ng/ml) D-dimer levels. The group with higher D-dimer values had lower mean minimal oxygen saturation (72.1 +/- 16.4 vs. 81.7 +/- 11.6%, p = 0.008) and a longer mean period of oxygen saturation below 90% (84.1 +/- 86.2 vs. 38.5 +/- 70.8 minutes, p = 0.032). There was no correlation of respiratory disturbance index and sleep architecture with D-dimer values. We concluded that sleep apnea syndrome is associated with fibrinolytic activity. Oxygen desaturation seems to be one of the mediatory factors in the putative connection between obstructive sleep apnea and hypercoagulability state. PMID- 16268469 TI - Drug interactions and risk of acute bleeding leading to hospitalisation or death in patients with chronic atrial fibrillation treated with warfarin. AB - Although drug interactions with warfarin are an important cause of excessive anticoagulation, their impact on the risk of serious bleeding is unknown. We therefore performed a cohort study and a nested case-control analysis to determine the risk of serious bleeding in 4152 patients (aged 40-84 years) with nonvalvular atrial fibrillation (AF) taking long-term warfarin (> 3 months). The study population was drawn from the UK General Practice Research Database. More than half (58%) of eligible patients used potentially interacting drugs during continuous warfarin treatment. Among 45 identified cases of incident idiopathic bleeds (resulting in hospitalisation within 30 days or death within 7 days) and 143 matched controls, more cases than controls took > or = 1 potentially interacting drug within the preceding 30 days (62.2% vs. 35.7%) and used > 4 drugs (polypharmacy) within the preceding 90 days (80.0% vs. 66.4%). Conditional logistic regression analysis yielded an odds ratio (OR) of 3.4 (95% confidence interval [CI]: 1.4-8.5) for the risk of serious bleeding in patients treated with warfarin and > or = 1 drugs potentially increasing the effect of warfarin vs. warfarin alone adjusted for polypharmacy, diabetes, hypertension, heart failure, and thyroid disease; the adjusted OR for the combined use of warfarin and aspirin vs. warfarin alone was 4.5 (95% CI: 1.1-18.1). We conclude that concurrent use of potentially interacting drugs with warfarin is associated with a 3 to 4.5-fold increased risk of serious bleeding in long-term warfarin users. PMID- 16268471 TI - Do patients followed in anticoagulation clinics for antiphospholipid syndrome meet criteria for the disorder? AB - Although specific criteria for diagnosing the antiphospholipid syndrome (APS) exist (the Sapporo Criteria), most physicians are not aware these include repeat testing and documentation of either a lupus anticoagulant or medium to high levels of anti-cardiolipin antibody. Incorrect diagnosis of APS may result in unnecessary long-term anticoagulation. The purpose of this study was to determine the clinical and serological characteristics of patients being treated for APS and concordance with published criteria. This cross-sectional study identified APS patients who were being treated with warfarin at one of three university based anticoagulation clinics. Levels of anticardiolipin antibody were classified as low-positive if abnormal but < 40 GPL/MPL units and medium/high-positive if > or = 40 units. Strength of meeting Sapporo criteria was graded as definite, possible, and not meeting criteria. Of 103 cases, 97 had clinical and laboratory data available. Only 10 cases (10%, 95% Confidence Interval 5 - 19) met criteria for definite APS, 16 (16%, 10 - 26) had a possible diagnosis, and 71 (73%, 63 - 81) did not meet criteria. Of 70 cases that had abnormal anticardiolipin antibody results, only 32 (46%, 34 - 58) had medium/high-positive levels. Repeat laboratory testing was performed in only 49 cases (51%, 40 - 61). We conclude that few patients treated forAPS met Sapporo criteria. Abnormal levels of anticardiolipin antibody were frequently in the low-positive range, and repeat testing was often absent. A quality improvement program that includes review of cases referred for chronic anticoagulation care is recommended to ensure appropriate testing and treatment of patients with suspected APS. PMID- 16268472 TI - ADAM 15 is an adhesion receptor for platelet GPIIb-IIIa and induces platelet activation. AB - Cell adhesion and proteolytic matrix degradation are central processes in atherosclerosis. Being a member of the family of ADAMs ("a disintegrin and metalloproteinase"), metargidin (ADAM15) combines a metalloproteinase domain and an RGD aminoacid sequence. We studied the potential role of ADAM15 as an adhesion receptor on endothelial cells and interactions between platelets and ADAM15 with respect to platelet adhesion, activation and thrombus formation. ADAM15 was found to be expressed on cultured endothelial cells (HUVEC). Platelet adhesion to immobilized recombinant ADAM15 was effectively enhanced under both static and high shear rate conditions reaching the maximum level of adhesion to fibrinogen. Consistently, platelet adhesion onto ADAM15 overexpressing endothelial cells was significantly increased. Adhesion to ADAM15 was reduced by blockade of GPIIb-IIIa using neutralizing anti-alpha(IIb)beta3 mAbs (7E3, 2G12), but not by anti alpha(v)beta3 (LM609). Soluble ADAM15 binds to activated but not to resting GPIIb IIIa. Moreover, platelets adherent to ADAM15 additionally attracted platelets under high shear rates indicating an initial role of platelet-ADAM15 interactions for thrombus formation. Furthermore, incubation of platelets with soluble ADAM15 showed a dose-dependent increase in secretion of CD62P and CD40L. ADAM15 is expressed on endothelial cells and can serve as an adhesion receptor for platelets via GPIIb-IIIa binding. Platelet adhesion to ADAM15 leads to platelet activation, secretion and promotes thrombus formation. Thus, ADAM15 may represent a novel target for antithrombotic strategies in cardiovascular pathologies. PMID- 16268473 TI - The decellularized porcine heart valve matrix in tissue engineering: platelet adhesion and activation. AB - An approach in tissue engineering of heart valves is the use of decellularized xenogeneic matrices to avoid immune response after implantation. The decellularization process must preserve the structural components of the extracellular matrix to provide a biomechanically stable scaffold. However, it is known that in vascular lesions platelet adhesion to extracellular matrix components occurs and platelet activation is induced. In the present study we examined the effects of a decellularized porcine heart valve matrix on thrombocyte activation and the influence of re-endothelialisation in vitro. Porcine pulmonary conduits were decellularized using Triton X-100, Na deoxycholate and Igepal CA-630 followed by a ribonuclease digestion. Cryostat sections of decellularized heart valves with and without seeding with human umbilical vein endothelial cells (HUVEC) were incubated with platelet rich plasma. Samples were either stained with fluorescent antibodies for CD41 and PAC I (recognizing the activated fibrinogen receptor) or fixed with glutaraldehyde. Thereafter, the samples were processed for laser scanning microscopy (LSM) or scanning electron microscopy (SEM). Examination by LSM showed numerous platelets with co-localized staining for CD41 and PAC-1 on the nonseeded decellularized heart valve matrix whereas after seeding with endothelial cells no platelet activation was detected. SEM revealed platelet adhesion and aggregate formation only on the surface of the non-seeded or partially denuded matrix specimens. We show in this study that the decellularized porcine matrix acts as a platelet activating surface. Seeding with endothelial cells effectively abolishes the platelet adhesion and activation and therefore is necessary to eliminate thrombogenicity in tissue engineered heart valves. PMID- 16268474 TI - Clopidogrel inhibits platelet-leukocyte adhesion and platelet-dependent leukocyte activation. AB - Clopidogrel is considered to be an important therapeutic advance in anti-platelet therapy. We investigated whether inhibition by clopidogrel results in a reduced capacity of platelets to adhere and stimulate pro-atherothrombotic and inflammatory functions in polymorphonuclear leukocytes (PMN) and in monocytes (MN). An eventual effect on these processes could further substantiate anti atherothrombotic properties of this drug. The effects of clopidogrel or of its active metabolite were investigated on ADP or thrombin receptor-induced platelet activation and on platelet-leukocyte interactions ex vivo in the mouse or in vitro in isolated human cells or whole blood, respectively. Clopidogrel inhibited platelet aggregation, expression of P-selectin, platelet-PMN adhesion and platelet-dependent ROS production in mouse PMN. Similarly pretreatment of human platelets with the active metabolite of clopidogrel in vitro resulted in a profound inhibition of platelet P-selectin expression, platelet-PMN adhesion and production of ROS by PMN. Pretreatment with the active metabolite of clopidogrel significantly impaired the ability of platelets to up-regulate the expression of TF procoagulant activity in MN, in a washed cell system. Moreover, the active metabolite of clopidogrel inhibited rapidTF exposure on platelet as well as on leukocyte surfaces in whole blood. By reducing platelet-dependent up-regulation of inflammatory and pro-atherothrombotic functions in leukocytes, clopidogrel may reduce inflammation that underlies the chronic process of atherosclerosis and its acute complications. PMID- 16268475 TI - Effect of nitroxyl on human platelets function. AB - There is a growing body of evidence on the role of nitric oxide (NO) in human platelet physiology regulation. Recently, interest has developed in the functional role of an alternative redox form of NO, namely nitroxyl (HNO/NO-), because it is formed by a number of diverse biochemical reactions. The aim of the present study was to comparatively analyze the effect of HNO and NO on several functional parameters of human platelets. For this purpose, sodium trioxodinitrate (Angeli's salt,AS) and sodium nitroprusside (SNP) were used as HNO and NO releasers, respectively. BothAS and SNP significantly inhibited platelet aggregation and ATP release induced by different agonists and adrenaline. AS or SNP did not modify the expression of platelet glycoproteins (Ib, IIb-IIIa, la-IIa, IV), whereas they substantially decreased the levels of CD62P, CD63 and of PAC-1 (a platelet activated glycoprotein IIb/IIIa epitope) after the stimulation with ADP. AS and SNP significantly increased cGMP accumulation in a 1H-[1,2,4]oxadiazolo [4,3-a] quinoxalin-1-one (ODQ)-sensitive manner. However, while L-cysteine reduced the effect of AS, it increased the effect of SNP on this parameter. Accordingly, a differential effect of L-cysteine was observed on the antiaggregatory effect of both compounds. In summary, these results indicate that HNO is an effective inhibitor of human platelet aggregation. PMID- 16268476 TI - Endocytosis and storage of plasma factor V by human megakaryocytes. AB - Factor V is an essential coagulation cofactor that circulates in plasma and platelet alpha-granules where it is stored complexed to multimerin I (MMRN1). To gain insights into the origin and processing of human platelet factor V, and factor V-MMRN I complexes, we studied factorV in cultured megakaryocytes. Factor V mRNA was detected in all megakaryocyte cultures. However, like albumin, IgG and fibrinogen, factorV protein was detectable only in megakaryocytes cultured with exogenous protein. The amount of factor V associated with megakaryocytes was influenced by the exogenous factorV concentration. Similar to platelet factor V, megakaryocyte factor V was proteolyzed and complexed with megakaryocyte synthesized MMRN1. With secretagogues, megakaryocytes released factor V, IgG, fibrinogen and MMRN1. Immunofluorescent and electron microscopy confirmed factorV uptake by endocytosis and its trafficking to megakaryocyte alpha-granules. These data provide direct evidence that human megakaryocytes process plasma-derived factor V into alpha-granules and generate factorV-MMRN I complexes from endogenously and exogenously synthesized proteins. PMID- 16268477 TI - Stereoselective inhibition of human platelet aggregation by R-138727, the active metabolite of CS-747 (prasugrel, LY640315), a novel P2Y12 receptor inhibitor. AB - CS-747 (Prasugrel, LY640315) is a thienopyridine antiplatelet prodrug that is metabolized to the thiol-containing active metabolite R-138727,which binds to and irreversibly inhibits the platelet P2Y12ADP receptor. R-138727 is composed of 4 stereo-isomers, (R, S)-, (R, R)-, (S, S)-, and (S, R)-isomers (the first letter for the configuration of a chiral center at the sulfur-bearing position and the second for that at the benzylic position). In the present study, we determined the stereoselectivity of P2Y12 antagonist effects by assessing the antagonism of the [3H]-2-MeS-ADP that binds to human P2Y12 receptors expressed in Chinese hamster ovary cells as an affinity assay, and by the inhibition of ADP-induced aggregation of washed human platelets as a functional assay. R-138727 and its 2 components, R-99224, a mixture of (R, S)- and (S, R)-isomers and R-100364, a mixture of (R, R)- and (S, S)-isomers, inhibited [3H]-2-MeS-ADP binding and platelet aggregation. The rank order of potency of these compounds were identical in both assays: R-99224>R-138727>> R-100364. Inhibition of ADP-induced platelet aggregation by R-138727 and R-99224 was concentration- and time-related. In experiments using the 4 single stereo-isomers, all isomers inhibited ADP-induced platelet aggregation, but the (R, S)-isomer was found to be the most potent, followed by the (R, R)-isomer. These in vitro studies indicate that R- 138727 is an effective antagonist of P2Y12 and potent inhibitor of ADP-induced platelet aggregation, and that these antiplatelet activities of R-138727 are largely dependent on its (R, S)-isomer. This suggests that the (R)-configuration of the reactive thiol group of the active metabolite of CS-747 is critical for P2Y12 and platelet inhibitory activities. PMID- 16268478 TI - A common ancestral glycoprotein (GP) 9 1828A>G (Asn45Ser) gene mutation occurring in European families from Australia and Northern Europe with Bernard-Soulier Syndrome (BSS). AB - Bernard-Soulier syndrome (BSS) is an extremely rare hereditary bleeding disorder, caused by mutations occurring in the Glycoprotein (GP) Ibalpha, GPIbbeta and GP9 genes that encode for the corresponding subunits of platelet GPIb-V-IX adhesion receptor complex. BSS has been reported in many populations, mostly behaving in an autosomal-recessive manner.While the great majority of BSS mutations are unique to a single individual or family, the GP9 1828A>G Asn45Ser mutation, which we have identified in an undocumented Australian Caucasian, has already been reported in multiple unrelated Caucasian families from various Northern and Central European countries. Haplotype analysis of 19 BSS patients from 15 unrelated Northern European families (including 2 compound heterozygote siblings from a British family previously published, and 17 1828A>G Asn45Ser homozygotes), showed that 14 of these BSS patients from 11 of the 1828A>G Asn45Ser homozygote families share a common haplotype at the chromosomal region 3' to the GP9 gene. Hence, the results suggest that the GP9 1828A>GAsn45Ser mutation in these families is ancient, and its frequent emergence in the European population is the result of a founder effect rather than recurrent mutational events. Association of the 1828A>G Asn45Ser mutation with variant haplotypes in 4 other Northern European BSS families raised the possibility of a second founder event, or rare recombinations in these families. Additional members from these 'atypical' lineages would need to be screened to resolve this question. PMID- 16268479 TI - Domain 5 of cleaved high molecular weight kininogen inhibits endothelial cell migration through Akt. AB - Domain 5 (D5) of cleaved high molecular weight kininogen (HKa) inhibits angiogenesis in vivo and endothelial cell migration in vitro, but the cell signaling pathways involved in HKa and D5 inhibition of endothelial cell migration are incompletely delineated. This study examines the mechanism of HKa and D5 inhibition of two potent stimulators of endothelial cell migration, sphingosine 1-phosphate (S1P) and vascular endothelial growth factor (VEGF), that act through the P13-kinase-Akt signaling pathway. HKa and D5 inhibit bovine pulmonary artery endothelial cell (BPAE) or human umbilical vein endothelial cell chemotaxis in the modified-Boyden chamber in response toVEGF or S1P. The inhibition of migration by HKa is reversed by antibodies to urokinase-type plasminogen activator receptor. Both HKa and D5 decrease the speed of BPAE cell migration and alter the morphology in live, time-lapse microscopy after stimulation with S1P or VEGF. HKa and D5 reduce the localization of paxillin to the focal adhesions after S1P and VEGF stimulation. To better understand the intracellular signaling pathways, we examined the effect of HKa on the phosphorylation of Akt and its downstream effector, GSK-3alpha HKa and D5 inhibit phosphorylation of Akt and GSK-3alpha after stimulation withVEGF and S1P. Inhibitors of Akt and P13-kinase, the upstream activator of Akt, block endothelial cell migration and disrupt paxillin localization to the focal adhesions after stimulation with VEGF and S1P. Therefore we suggest that HKa through its D5 domain alters P13-kinase-Akt signaling to inhibit endothelial cell migration through alterations in the focal adhesions. PMID- 16268480 TI - Opposite relationship between circulating soluble CD14 concentration and endothelial function in diabetic and nondiabetic subjects. AB - Recent prospective studies indicate endothelial dysfunction and increased risk for cardiovascular events in patients with serological evidence of multiple infections. Soluble CD14 (sCD 14) plays a key role in the neutralization of lipopolysaccharide (LPS), a well-established bacterial product inducing endothelial dysfunction. Insulin resistance was recently identified as a significant factor influencing circulating sCD 14 concentration. Thus, we investigated the association of circulating sCD14 and endothelial dysfunction in subjects with well-established insulin resistance (patients with type 2 diabetes, n = 40) compared to control non-diabetic subjects (n = 100). To further explore the underlying mechanisms, we also analysed C-reactive protein and circulating NO2-/NO3- and cyclic GMP in the diabetic group. Serum sCD 14 concentration (ELISA) was found to be differently associated with endothelium-dependent vasodilatation (EDVD, high-resolution ultrasound) in diabetic and non-diabetic subjects. In nondiabetic subjects, serum sCD14 and C-reactive protein correlated negatively with EDVD (r = -0.21, p = 0.03, and r = -0.21, p = 0.03, respectively). In a partial correlation analysis, these associations remained significant after controlling for age and weight (sCD 14 and EDVD, r = -0.23, p = 0.023; C-reactive protein and EDVD, r = -0.21, p = 0.03; sCD14 and C-reactive protein, r = 0.30, p = 0.002). In contrast, sCD 14 was positively associated with EDVD in type 2 diabetic patients (r = 0.37, p = 0.019,). Interestingly, sCD14 was also associated with NO2-/NO3- in this group (r = 0.62, p = 0.001, n = 22). EDVD also correlated with cyclic GMP (r = 0.47, p = 0.03, n = 22). In summary, circulating sCD 14 is associated with endothelial function. While in non-diabetic subjects sCD14 behaves as an acute phase reactant, its role in type 2 diabetic patients should be further clarified. These findings need to be confirmed in further studies with larger number of patients. PMID- 16268481 TI - Tyrosine phosphorylation and the small GTPase rac cross-talk in regulation of endothelial barrier function. AB - Endothelial barrier function depends on the integrity of intercellular adherens junctions controlled by the association of VE-cadherin/catenin complex with cortical actin filaments. Both tyrosine phosphorylation/dephosphorylation of junctional proteins and actin reorganization mediated by rho-GTPases regulate barrier function but the relationship between these regulatory mechanisms is unclear. Here we studied the effects of factors increasing protein tyrosine phosphorylation, pervanadate (PV) and VEGF, on distribution of VE-cadherin, F actin polymerization and transendothelial electrical resistance (TER) in human umbilical vein endothelial cells (HUVECs). Changes in protein tyrosine phosphorylation of cytoplasmic and junctional proteins, as well as the activity of rho-GTPase racl, were also measured. We report for the first time that PV and VEGF induced a rapid transient increase in endothelial barrier function accompanied by rac1 activation, a differentiated tyrosine phosphorylation of the VE-cadherin/catenin complex, recruitment of actin filament to cell junctions and ruffle formation. A sustained decrease in endothelial barrier function was observed at later times of PV and VEGF treatment. Expression of dominant negative rac1, N17rac1 abolished the barrier-enhancing effects of PV and VEGF, while the sustained decrease in barrier function was unaffected. These observations bring into focus early short-term effects of protein tyrosine phosphorylation in cells, often overshadowed by more pronounced and long-lasting later effects and may play an important role in the regulation of endothelial barrier function. PMID- 16268482 TI - Congenital erythropoietin over-expression causes "anti-pulmonary hypertensive" structural and functional changes in mice, both in normoxia and hypoxia. AB - Acute alveolar hypoxia causes pulmonary vasoconstriction that matches lung perfusion to ventilation to optimize gas exchange. Chronic alveolar hypoxia induces pulmonary hypertension, characterized by increased muscularization of the pulmonary vasculature and right ventricular hypertrophy. Elevated erythropoietin (EPO) plasma levels increase hematocrit and blood viscosity and may affect structure and function of the pulmonary circulation. To differentiate between the direct effects of hypoxia and those linked to a hypoxia-induced increase in EPO/hematocrit levels, we investigated the lung vasculature in transgenic mice constitutively over-expressing EPO (termed tg6) upon exposure to normoxia and chronic hypoxia. Despite increased hematocrit levels (approximately 0.86),tg6 mice kept in normoxia did not develop selective right ventricular hypertrophy. The portion of vessels with a diameter of 51-95 microm and >155 microm was increased whereas the portion of small vessels (30-50 microm) was decreased. Pulmonary vascular resistance and the strength of hypoxic vasoconstriction measured in isolated perfused lungs were decreased. Vasoconstrictions induced by the thromboxane mimetic U46619 tended to be reduced. After chronic hypoxia (FiO2 = 0.10, 21 days), vascular resistance and vasoconstrictor responses to acute hypoxia and U46619 were reduced in tg6 mice compared to wildtype controls. Chronic hypoxia increased the degree of pulmonary vascular muscularization in wildtype but not in tg6 mice that already exhibited less muscularization in normoxia. In conclusion, congenital over-expression of EPO exerts an "anti pulmonary hypertensive" effect, both structurally and functionally, particularly obvious upon chronic hypoxia. PMID- 16268483 TI - Ethanol inhibits pulse pressure-induced vascular smooth muscle cell migration by differentially modulating plasminogen activator inhibitor type 1, matrix metalloproteinase-2 and -9. AB - We investigated the effect of ethanol on the pulse pressure-induced expression of PAI-1 and MMP-2/9 in human smooth muscle cells (SMC). Human SMC were exposed to static or pulse pressure (25 mL/min; pulse pressure 106/50 mm Hg) conditions for 24 h in the absence or presence of ethanol (0.1-100 mM). SMC migration was then measured by Transwell migration assay. SMC exposed to pulse pressure demonstrated a significant increase in PAI-1 mRNA and protein expression (approximately 4-fold and approximately 3-fold) concomitant with a 3- and 8-fold increase in MMP-2 and MMP-9 protein, respectively. Ethanol dose-dependently inhibited the pulse pressure-induced SMC migration with complete inhibition observed at 20 mM. There was no effect of ethanol on basal PAI-1 or MMP-2/9 in SMC under static conditions. However, ethanol significantly enhanced the pulse pressure-induced PAI-1 mRNA and protein expression (2.2 +/- 0.52 fold and 2.5 +/- 0.27 fold, for 10 mM), respectively. In contrast, ethanol dose-dependently inhibited the pulse pressure-induced increases in MMP-9 protein and pro-MMP-9 activity and to a lesser extent MMP-2 mRNA and protein and pro-MMP-2 activity, with significant inhibition observed at 1 mM. These data provide a molecular mechanism mediating the inhibitory effect of ethanol on pulse-pressure-induced SMC migration and may be relevant to the cardioprotective effects of ethanol in vivo. PMID- 16268484 TI - Interleukin-1 gene cluster variants and abdominal aortic aneurysms. AB - Inflammation is a key factor in the pathogenesis of abdominal aortic aneurysms (AAA). Interleukin 1 (IL-1), a fundamental regulator of the inflammatory cascade, has been shown to be involved in this process. Several functional polymorphisms in the IL-1 gene cluster are known. In this matched case-control study, we investigated a potential association between six genetic variants in IL-1 and IL 1 receptor antagonist (IL-1 RN) with AAA. We enrolled 405 individuals, 135 consecutive patients with AAA were individually age- and sex-matched to 270 patients with coronary artery disease (CAD). Traditional cardiovascular risk factors and IL-1 genotypes were determined, and the distribution of six single nucleotide polymorphisms were compared between patients and controls by multivariable conditional logistic regression analysis: IL-1A (-889) C>T, IL-1A (+4845) G>T, IL-1B (-511) C>T, IL-1B (-31) C>T, IL-1B (+3954) C>T and IL-1RN (+2018) C>T. IL-1A (-889) C>T and IL-1A (+4845) G>T (kappa 0.98,95% CI 0.96 to 1.00), and IL-1B (-511) C>T and IL-1B (-31) C>T (kappa 0.98, 95% CI 0.96 to 1.00) were closely linked, therefore IL-1A (-889) C>T and IL-1B (-31) C>T were not considered for further analyses. None of the 4 remaining polymorphisms showed a significant association with AAA: IL-1RN (+2018) C>T (p = 0.061), IL-1B (+3954) C>T (p = 0.51), IL-1B (-511) C>T (p = 0.61) and IL-1A (+4845) G>T (p = 0.81). No significant first-degree interactions between the genetic variants and AAA were detected. In conclusion,these six genetic variants in the interleukin- I gene cluster do not seem to play a clinically relevant role in the pathogenesis of AAA, although we cannot rule out the existence of higher degree gene-gene or gene environment interactions. PMID- 16268485 TI - CpG methylation of the PAI-1 gene 5'-flanking region is inversely correlated with PAI-1 mRNA levels in human cell lines. AB - The physiological and pathophysiological functions of PAI-1 are related to its expression by specific cell types in normal and diseased tissues. We analysed the contribution of DNA methylation to the variation in PAI-1 mRNA levels in five cell lines. We found varying frequencies of methylation of 25 CpGs in the 805/+152 region of the PAI-1 gene in Bowes, MCF-7 and U937 cells, while little or no methylation was detected in Hep2 and HT- 1080 cells. The methylation frequency was inversely correlated with PAI-1 mRNA level within its 20-fold range in Bowes, MCF-7, U937,and Hep2 cells, while the lack of methylation in both Hep2 and HT 1080 cells suggested another mechanism behind the 150-fold higher level in HT- 1080 cells than in Hep2 cells. However, all cell lines exhibited a high frequency of methylation of 10 CpGs in a CpG island at about--1800. Treatment with 5-aza-2' deoxycytidine led up to circa a 40-fold increase in the PAI-1 mRNA level and a strong decrease in the frequency of methylation in the -805/+152 region in Bowes, MCF-7 and U937. The histone deacetylase inhibitor trichostatin A induced a several fold increase of the PAI-I mRNA level in cells with a high methylation frequency of the -805/+152 region. As compared with matched normal tissue, three samples of oral squamous cell carcinomas displayed decreased frequencies of methylation of the PAI-1 5' flanking region and increased levels of PAI-1 mRNA. These results for the first time implicate DNA methylation and histone acetylation in regulation of the PAI-1 gene, and indicate that without proper CpG islands in 5'-flanking region,trancription may be regulated by methylation of less dense CpGs in the 5'-flanking region rather than methylation of upstream CpG island. PMID- 16268486 TI - Rapid identification of female haemophilia A carriers with deletions in the factor VIII gene by quantitative real-time PCR analysis. AB - Large deletions of the factorVIII gene account for approximately 5% of severe haemophilia A patients. Although deletions are readily detectable in males, the identification of heterozygosity in possible carriers of these families still constitutes a challenge. In order to identify a deleted allele over the background of the normal allele in these carriers, we developed a rapid real-time quantitative PCR approach by means of LightCycler technology and SYBR green I for monitoring product formation. The method was applied to families with independent deletions (one in exon 14 and the other in exons 23-24) of the Factor VIII gene, thereby allowing a reliable determination of carrier or non-carrier status. The method is extremely versatile and can be adapted to other deletions within the factorVIII gene as well as to other diseases whose molecular pathology consists of deletions or duplications. PMID- 16268488 TI - Immediate type hypersensitivity after injection of nadroparin (Fraxiparin). PMID- 16268487 TI - Variability of anti-beta2 glycoprotein I antibodies measurement by commercial assays. AB - The aim of this study was to evaluate the agreement in assay results between commercial kits for the measurement of anti-beta2glycoprotein I antibodies. Ten manufacturers provided one IgG and one IgM kit to three testing centres. Samples from patients with primary (n = 13) or secondary (n = 3) antiphospholipid syndrome (APS), from lupus patients without APS features (n = 6) and from normal individuals (n = 2) were tested in the three centres according to manufacturers' instructions. Dilutions in normal serum of a pool made from positive patients' samples (Forum Calibrators) and dilutions of humanized monoclonal antibodies (MoAbs) were used as additional calibrators. The calibration curves obtained with each calibrator differed widely between kits. The rate of positivity of patients' samples varied from 7 to 16 for IgG and from 2 to 17 for IgM, depending on the kit. Perfect agreement occurred in 12/22 samples for IgG and 5/22 samples for IgM. Samples from normals were found negative by all kits. Between kits, cutoff values varied up to five fold when expressed in Forum Calibrators arbitrary units and up to three fold when expressed in MoAbs equivalents. Examination of discrepant samples indicated that about half of the discrepancies, scoring 8:2 and 9:1, involved the same few kits. In highly discrepant samples, some kits appeared as high responders as compared to others. In conclusion, with the exception of a few kits, agreement in assay results was acceptable. In conclusion, additional efforts are however necessary, especially concerning the way to assess the cutoff point and the adoption of a reference calibrator, in order to improve standardization of the assays. PMID- 16268490 TI - Identification of the novel signal peptide mutation, antithrombin-Siriraj causes type-I antithrombin deficiency in thai patients. PMID- 16268489 TI - First report of two independent point factorVIII mutations in a family with haemophilia a: a word of caution for carrier diagnosis. PMID- 16268492 TI - Transitory reduction of platelet aggregation with the use of etonogestrel implant in healthy women. PMID- 16268491 TI - Distal embolization during angioplasty for acute myocardial infarction: is pharmacological protection enough? PMID- 16268493 TI - Treatment of acute central retinal artery occlusionwith the platelet glycoprotein IIb/IIIa receptor inhibitor tirofiban. PMID- 16268494 TI - The thrombomodulin-1208/-1209delTT gene promoter polymorphism does not affect basal or LPS-dependent monocyte TM mRNA transcription in healthy volunteers. PMID- 16268495 TI - Rebuttal to: strong lupus anticoagulant can influence the prothrombin time INR falsifying the follow up of oral anticoagulation. PMID- 16268496 TI - In vivo effect of Chianti red wine on tissue factor, tissue factor pathway inhibitor and homocysteine levels. PMID- 16268497 TI - Vitamin K epoxide reductase (VKORC1) genetic polymorphism is associated to oral anticoagulant overdose. PMID- 16268498 TI - [Role of P-glycoprotein in pharmacokinetics and its clinical implications]. PMID- 16268499 TI - [Significant role of transporters in drug hepatobiliary transport]. PMID- 16268500 TI - [Inhibitory effects of chloride channel blockers NPPB on proliferation of human glomerular mesangial cells]. AB - AIM: To investigate the effects of NPPB, a chloride channel blocker, on proliferation of mesangial cells. METHODS: Cell proliferation was determined by measuring cell number and 3H-thymidine incorporation. The LDH activity released from these cells was measured as evaluation of cell viability. The phase of cell cycle was detected by flow cytometry. RESULTS: Cell proliferation assays showed that treatment with both NPPB (50 and 25 micromol x L(-1)) and in hypertonic media (100% increased osmolarity with D-mannitol ) significantly reduced the number of human MC and 3H-thymidine incorporation in a dose-dependent manner. But the LDH activity was not significantly altered in the treatment with 50 micromol x L(-1) NPPB. Flow cytometry experiments showed that 50 and 25 micromol x L(-1) NPPB arrested (84.2 +/- 2.4) % and (80.8 +/- 2.9) % of cells at G0/G1 stage, versus (70.5 +/- 1.4) % of control cells. Conclusion NPPB suppresses cell proliferation and produces growth arrest at G0/G1 phase in human MC by a mechanism probably associated with changes in cell volume. PMID- 16268502 TI - [Influence of cytochrom P450 CYP2C9 polymorphism on the pharmacokinetics of tolbutamide metabolism using oligonucleotide genotyping microarray]. AB - AIM: To investigate the influence of cytochrom P450 CYP2C9 polymorphism on the pharmacokinetics of tolbutamide. METHODS: An oligonucleotide microarray was designed and fabricated to genotype the CYP2C9 accurately and quickly. 137 healthy volunteers were genotyped with the array to investigate the frequency of CYP2C9 functional SNPs. Moreover, 1 homozygous mutant, 9 heterozygous and 10 wild genotypes subjects in the assay were selected randomly and sequenced directly. After orally taking tolbutamide, blood samples and urine samples were collected, and their pharmacokinetics was studied with HPLC. RESULTS: CYP2C9 *1/*3 were found in 9 of 137 volunteers, CYP2C9 *3/*3 in only one, others were all CYP2C9 *1/*1 wild types. CYP2C9 *2, CYP2C9 *4 and CYP2C9 *5 alleles were not detected. Direct sequencing of the purified PCR products of the heterozygotes, mutant homozygotes and ten wild type individuals gave a corresponding result to that genotyped by microarray. Pharmacokinetic outcome showed that the individuals with CYP2C9 *1/*3 or CYP2C9 *3/*3 had slower metabolic elimination of tolbutamide than those with CYP2C9 *1/*1. CONCLUSION: CYP2C9 genetic polymorphism has a significant influence on the pharmacokinetics of tolbutamide. Pharmacogenomic study will be helpful in guiding rational and individualized medication. Key words: tolbutamide; cytochrom P450 CYP2C9; allele; single nucleotide polymorphism; genotyping PMID- 16268501 TI - [Protective effects of ligustrazine against photoreceptor cell injury induced by N-methyl-N-nitrosourea and its mechanism]. AB - AIM: To study the protective effect of ligustrazine against photoreceptor cell injury induced by N-methyl-N-nitrosourea (MNU) in Sprague-Dawley (SD) rats. METHODS: Ligustrazine injections of different doses were injected intraperitoneally into 47-day female SD rats once a day and a single intraperitoneal injection of MNU 60 mg x kg(-1) was given to 50-day rats. At different intervals after MNU treatment,the animals were sacrificed. The apoptotic index of photoreceptor cells was calculated by TUNEL labeling at 24 h following MNU treatment; peripheral retinal damage was evaluated based on retinal thickness at the d 7 after MNU treatment, and the expression of c-jun and c-fos genes was detected by RT-PCR technique. RESULTS: Ligustrazine injection could remarkably increase total thickness of peripheral retina and decrease apoptotic index of photoreceptor cells induced by MNU in a dose-dependent manner. Compared with MNU-treated rats, the gene expression of c-jun and c-fos was time dependently down-regulated in ligustrazine-treated group. CONCLUSION: Ligustrazine injection partially protects against MNU-induced retinal damage by down-modulating the expression of c-jun and c-fos genes to inhibit apoptosis of photoreceptor cells. PMID- 16268503 TI - [Effect of salidroside on mitochondria injury induced by sodium azide]. AB - AIM: To study the protective effect of salidroside on mitochondria injury induced by sodium azide. METHODS: Human neuroblastoma SH-SY5Y cells were exposed to sodium azide with different concentration of salidroside, then cell viability was measured by thiazolyl blue (MTT) method and mitochondrial membrane potential (MMP) was detected by JC-1 method. Protective effect of salidroside against disfunction of mitochondria induced by sodium azide was detected by resazurin method. RESULTS: After exposing to 64 mmol x L(-1) sodium azide for 4 hours, cell viability and MMP of SH-SY5Y cells significantly decreased. When pretreated with salidroside, the cell damage was greatly reduced and the mitochondrial membrane potential was maintained. Furthermore, salidroside can protect function of rat brain mitochondria against damage induced by sodium azide. CONCLUSION: Salidroside was demonstrated to play an important role in improving the function of mitochondria. PMID- 16268504 TI - [Protection of oxyphenamone against cardiac arrest-reperfusion injury of isolated rat hearts]. AB - AIM: To ascertain the protective effect of oxyphenamone, a novel inodilator, against myocardial ischemia-reperfusion injury. METHODS: A model of global myocardial ischemia-reperfusion injury (I-R) was established by arresting the infusion to heart 40 min followed by reperfusion 30 min in the preparations of rat Langendorff' s hearts. The protective effects of oxyphenamone were evaluated by the cardiac function, the activity of creatine phosphokinase (CPK) in coronary efflux, the myocardial energy metabolism and antioxidation, the membrane fluidity, the activity of ATPase, the calcium content and ultrastructure of mitochondria. RESULTS: Administration of oxyphenamone to the infused heart before cardiac arrest and during reperfusion ameliorated the decreases of myocardial contractile force and coronary flow and the increase of the activity of CPK in coronary efflux induced by cardiac arrest-reperfusion (A-R). At the concentration of 1-10 micromol x L(-1) oxyphenamone dose-dependently antagonized the decrease of myocardial adenosine triphosphate (ATP) and creatine phosphate (PCr) induced by A-R, increased the activities of myocardial superoxide dismutase (SOD) and glutathione peroxidase (GSH-px), decreased mitochondrial malondialdehyde (MDA) content and increased membrane fluidity, glutathione (GSH) content and the activities of GSH-px and ATPase of mitochondria. The parameters mentioned above even maintained at normal level when high concentration of oxyphenamone (10 micromol x L(-1)) was applied. Oxyphenamone also antagonized the mitochondrial calcium overload and the ultrastructure damage of mitochondria induced by A-R obviously. Addition of oxyphenamone (1-100 micromol x L(-1)) to the system of Fe2+-cysteine or Fe2+-H2O2, which could produce oxygen free radicals, decreased MDA content and increased GSH and membrane fluidity of mitochondria significantly. CONCLUSION: With the results of examinations of the cardiac physiological function, myocardial energy metabolism and antioxidation and the calcium content and ultrastructure of mitochondria, it is indicated that oxyphenamone could protect the isolated rat heart against cardiac arrest reperfusion injury markedly and the mechanism of its action may be related to the antioxidative effect of this agent. PMID- 16268505 TI - [Synthesis and antitumor activity of arylsubstituted imidazolin-2-one derivatives]. AB - AIM: To design and synthesize new arylsubstituted imidazolin-2-one analogues as antitumor compounds. METHODS: Arylsubstituted imidazolin-2-ones were prepared by condensation the appropriate omega-amino-acetophenone hydrochloride with arylisocyanate in toluene. The target compounds prepared in this study were tested for cytotoxicity against PC-3, A549, HO-8910, Hela, HL60, K562 and HL60R cancer cell lines, and mechanism of one of the products 4y was further evaluated with its mechanium. RESULTS: Thirty-six new compounds were synthesized and confirmed by 1H NMR, MS and elemental analysis. One of the synthesized products, compound 4y, displayed an encouraging selective activity against HL60 cells, and it was partlydue to the cell cycle arrest and cell apoptosis. CONCLUSION: Compound 4y is worthy to be intensively studied. PMID- 16268506 TI - [The biotransformation of kaempferitrin by human intestinal flora]. AB - AIM: To study the biotransformation of kaempferitrin, a major chemical principle of the fruits of Siraitia grosvenori (Swingle) C. Jeffery, with human intestinal flora. METHODS: The kaempferitrin was incubated with human intestinal flora. The biotransformation products were isolated and purified by chromatographic methods and the structures were determined by spectroscopic techniques. RESULTS: Kaempferitrin was converted into kaempferol 3-O-alpha-L-rhamnoside (afzelin, I) , kaempferol 7-O-alpha-L-rhamnoside (II), kaempferol (III) and p-hydroxybenzoic acid (IV) by human intestinal flora. rhamnoside (II), kaempferol (III) and p hydroxybenzoic acid (IV) by human intestinal flora. CONCLUSION: The structure of kaempferitrin can be biotransformatedly converted by human intestinal flora. PMID- 16268507 TI - [Isolation and characterization of phenylethanoid glycosides from Clerodendron bungei]. AB - AIM: To study the chemical constituents from Clerodendron bungei Steud. METHODS: The compounds were isolated and purified by various chromatographic techniques and identified by their physicochemical properties and spectral data. RESULTS: Ten phenylethanoid glycosides were isolated and identified as clerodendronoside (1), acteoside (2), isoacteoside (3), cistanoside C (4), jionoside C (5), leucosceptoside A (6), cistanoside D (7), campneoside I (8), campneoside II (9), cistanoside F (10). CONCLUSION: Compound 1 is a new phenylethanoid glycoside, while compounds 4-10 are obtained from this plant for the first time. PMID- 16268508 TI - [Molecular identification of medicinal plants: Dendrobium chrysanthum, Dendrobium fimbriatum and their morphologically allied species by PCR-RFLP analyses]. AB - AIM: To establish a simple method for molecular identification of original plants of D. chrysanthum and D. fimbriatum using molecular marker rDNA ITS region. METHODS: Restriction patterns of ITS fragments were obtained using PCR-RFLP method. The PCR products of D. chrysanthum and its morphologically allied species were digested at 37 degrees C by Cla I and Apa LI, those of D. fimbriatum and its morphologically allied species were digested by Sph I. RESULTS: D. chrysanthum, D. fimbriatum and their morphologically allied species could be identified by predicted restriction profiles of PCR-RFLP. The botanical origin of twenty-five fresh samples of "Shihu" collected in markets was identified by this method. CONCLUSION: The results showed that PCR-RFLP analysis of the rDNA ITS region is a feasible, simple and inexpensive method for determining the botanical origin of the traditional Chinese medicine "Shihu". PMID- 16268509 TI - [LC-ESI-MS metabolic profiling analysis of taxanes from the extracts of Taxus chinensis cell cultures]. AB - AIM: To develop a rapid analytical method for small amount biological samples of taxanes by using liquid chromatography coupled with electrospray ionization tandem mass spectrometry (LC-ESI-MS) in small amount biological samples. METHODS: A solution containing five given taxane constituents and the extract from cell cultures of Taxus chinensis were analysed separately. According to the performance of the given taxanes in ESI-MS/MS, run parameters of the mass spectrometer were optimized. Positive and negative electrospray modes were employed to simultaneously scan the cell cultures sample, and the full ion chromatogram and the molecular weight of individual peak were obtained. The qualitative analysis of taxanes was achieved by comparison of the retention time and molecular weight with those of the reference substances or was based on the interpretation of the MS/MS spectra of the analytes and the knowledge of the concerning genetic backgrounds of taxanes published in literatures. RESULTS: The taxanes with several acetyl substituents tended to produce ammonium adduct ions peak, while multi-hydroxy taxanes were subject to give protonized molecular ion peaks in positive ion ESI-MS. Thirteen taxanes in cell samples were assigned. Eight compounds of them were identified to be 1 -acetyl-5, 7, 10-deacetyl baccatin I (DAB-I, 1) , baccatin III (B-III, 3), baccatin VI (B-VI, 8), taxol (9), yunnanxane (10 ), taxuyunnanine C (Tc, 11), sinenxane B (12), sinenxane C (13), separately. For the other five constituents, character of taxane and the number of substituents were deduced. CONCLUSION: The results confirm the feasibility of characterizing taxanes in biological samples by LC-ESI-MS analysis. The analytical methodology provided a rapid, conventional and reliable tool to study metabolic profiling of taxanes for structural elucidation in taxol biosynthesis. PMID- 16268510 TI - [HPLC-electrospray ionization ion trap tandem mass spectrometry analysis of oxymatrine and its metabolites in rat urine]. AB - AIM: To identify the main metabolites of oxymatrine (OMT) in rats. METHODS: To optimize the conditions of LC/ESI-ITMS' chromatograms and spectra by oxymatrine and matrine (MT), and summarize their ionization and cleavage rules in ESIMS, then serving as the basis for the metabolite analyses of oxymatrine in rats. To collect the 0-24 h urine samples of the rats after ip 40 mg x kg(-1) oxymatrine, the samples were enriched and purified through C18 solid-phase extraction cartridge. The purified samples were analyzed by LC/ESI-ITMS. The structures of OMT metabolites were identified according to their retention times and ESI-ITMSn rules. RESULTS: Six phase I metabolites and the parent drug OMT were found in the rat urine, and the main metabolite was MT. No phase II metabolites were found. CONCLUSION: The developed LC/ESI-ITMSn methods to identify the metabolites of oxymatrine in rats is not only simple and rapid but also sensitive and specific. This technology is one of the most efficient methods for the analysis of drug metabolites. PMID- 16268511 TI - [Stereoselective pharmacokinetics of tetrahydropalmatine in rats]. AB - AIM: To investigate the stereoselective pharmacokinetic process of tetrahydropalmatine (THP) in rats. METHODS: The concentrations of tetrahydropalmatine enantiomers in rat plasma were determined by coupled achiral and chiral HPLC method. The differences in plasma concentrations and pharmacokinetic parameters between the two enantiomers were compared by paired t test. RESULTS: The plasma levels of l-THP were always higher than those of d-THP in eight rats. There was significant difference between the main pharmacokinetic parameters of the two enantiomers. CONCLUSION: Tetrahydropalmatine showed significant stereoselective pharmacokinetics in rats after an ig dose of the racemate. PMID- 16268512 TI - [Polarographic behavior and determination of glimepiride]. AB - AIM: To establish a polarographic method of parallel catalytic hydrogen wave for determination of glimepiride. METHODS: The catalytic wave of glimepiride in the presence of K2S2O8 was used to improve the analytical sensitivity. The rapid determination of glimepiride was done by linear single sweep polarography. RESULTS: The catalytic hydrogen wave of glimepiride was measured at ca. -1.36 (vs SCE) in 0.09 mol x L(-1) Na2B4O7-KH2PO4 (pH 6.24 +/- 0.1) supporting electrolyte. When 1.0 x 10(-2) mol x L(-1) K2S2O8 was present, the current increased by 25 times, and the peak potentioal was unchanged, producing a more sensitive parallel catalytic hydrogen wave. The peak current of the parallel catalytic hydrogen wave was rectilinear to the glimepiride concentration in the range 1.0 x 10(-7) - 4.2 x 10(-5) mol x L(-1) (r = 0.9990, n = 9). The detection limit was 5.0 x 10(-8) mol x L(-1). CONCLUSION: The proposed method could be applied to the determination of glimepiride in pharmaceuticals without preliminary separation. PMID- 16268513 TI - Enhancement of systemic and CNS delivery of meptazinol hydrochloride by intranasal administration to rats. AB - AIM: To investigate the extent of systemic absorption and uptake of meptazinol (MEP) hydrochloride in cerebrospinal fluid (CSF) after intranasal administration on rats and compare with oral administration. METHODS: CSF samples were collected by a serial sampling method. The concentration of MEP in the biological samples was measured by HPLC with fluorescence detector. RESULTS: Rapid and significant levels of MEP in plasma and CSF can be achieved after nasal administration whereas the oral administration resulted in considerably lower drug concentrations. AUC in plasma and CSF from the nasal route are 7.375 and 15.6 folds compared with those of the oral route, respectively. CONCLUSION: Intranasal MEP is able to show quick absorption and improve the bioavailability, which could be a promising alternative to oral administration. PMID- 16268514 TI - [Preparation of silymarin proliposomes and its pharmacokinetics in rats]. AB - AIM: To study the preparation of silymarin proliposomes. To study its physicochemic properties, its pharmacokinetical characteristics and bioavailability in rats after oral administration. METHODS: Silymarin proliposomes were prepared by film-deposition on carriers. When the proliposomes were contacted with water to form liposome suspensions, the tests of physicochemical properties including encapsulation efficiency, particle size and stability of the formed liposome suspensions were determined by HPLC, laser particle-sizer and etc. The concentrations of non-conjugated and overall silymarin in plasma of rats and their pharmacokinetic behaviors after oral administration were studied by RP-HPLC. The pharmacokinetic parameters were computed by software program 3P97. RESULTS: The encapsulation efficiency of silymarin liposomes could be more than 90%, with an average particle size of about 238.8 nm and a very good stability. The high bioavailability of silymarin proliposomes could be gotten by oral administration. CONCLUSION: Compared with silymarin, silymarin proliposome is a stable and easily industrialized preparation and did enchance the gastrointestinal absorption of silymarin. PMID- 16268516 TI - Access to isocarbacyclin derivatives via substrate-controlled enolate formation: total synthesis of 15-deoxy-16-(m-tolyl)- 17,18,19,20-tetranorisocarbacyclin. AB - [reaction: see text] We describe a convergent and flexible synthesis of 15-deoxy 16-(m-tolyl)-17,18,19,20-tetranorisocarbacyclin (15-deoxy-TIC), a simple isocarbacyclin derivative. The synthesis takes advantage of two key step reactions: a regioselective deprotonation of the described ketone under substrate control which is then trapped, as the enol triflate, to generate the C6-C9alpha endocyclic double bond, followed by an sp2-sp3 Pd-catalyzed cross-coupling reaction (C5-C6) with a suitable primary alkyl Grignard reagent. Introduction of the C13-C14 (E)-double bond in the omega-side chain is performed by the Julia Kocienski olefination. PMID- 16268515 TI - [In vitro transdermal permeation of lidocaine-dodecanol binary eutectic system]. AB - AIM: To study the effect of lidocaine-dodecanol binary eutectic system on the transdermal permeation of lidocaine. METHODS: Binary eutectic mixture of different proportions of lidocaine and dodecanol were prepared and the patch containing the binary eutectic mixture was developed. The solubilities of pure lidocaine and lidocaine from the binary eutectic system were determined in pH 7.9 phosphate buffer. The transdermal flux of lidocaine from the patches containing the binary eutectic system and pure lidocaine were measured using Franz-type single diffusion cell. RESULTS: The melting point of the lidocaine-dodecanol binary eutectic system was markedly lower than that of pure lidocaine. The steady state transdermal flux of lidocaine from the patch of the binary eutectic system was six times as much as that of pure lidocaine patch. CONCLUSION: The lidocaine dodecanol binary eutectic system could produce high thermodynamic activity of the drug and the high driving force for transdermal permeation of lidocaine. PMID- 16268517 TI - N-aryl-9-amino-substituted acridizinium derivatives as fluorescent "light-up" probes for DNA and protein detection. AB - [reaction: see text] N-Arylamino-substituted acridizinium (benzo[b]quinolizinium) derivatives are almost nonfluorescent in water or organic solvents; however, upon addition of calf thymus DNA or bovine serum albumin the fluorescence intensity increases by a factor of 10 to 50. Thus, these dyes exhibit ideal properties to be used as DNA- and protein-sensitive "light-up probes". PMID- 16268518 TI - Single-site catalysis by bimetallic zinc calixarene inclusion complexes. AB - [reaction: see text] Bimetallic alkylzinc calixarene inclusion complexes were prepared and used in the ring-opening polymerization of L-lactide. Polymers with high molecular weight and a low degree of polydispersity were obtained. 1H NMR and diffusion NMR experiments suggest that a single-site mechanism is operative. PMID- 16268520 TI - Spiro-configured bifluorenes: highly efficient emitter for UV organic light emitting device and host material for red electrophosphorescence. AB - [structure: see text] Remarkable UV OLED devices using novel spiro-configured bifluorenes as UV emitters have been successfully achieved with external quantum efficiencies up to 2.6-3.1% and emission wavelengths as short as 370 nm. In addition, these large-gap materials have been shown to be effective hosts for red phosphorescent emitters in phosphorescent OLEDs. This result represents the first example that pure hydrocarbon molecules can serve as an efficient host for red electrophosphorescence devices. PMID- 16268519 TI - Studies toward a marine toxin immunogen: enantioselective synthesis of the spirocyclic imine of (-)-gymnodimine. AB - [reaction: see text] An enantioselective Diels-Alder reaction catalyzed by an Evans' copper-bis(oxazoline) complex was utilized to construct a highly functionalized spirolactam, a key intermediate in our projected total synthesis of the marine toxin, gymnodimine. Additional transformations, including a mild N tosyl group deprotection, afforded a keto spirocyclic imine moiety, the proposed pharmacophore of gymnodimine. Thus, the prepared ketone is a potentially useful intermediate for conjugation to provide an immunogen for eventual monitoring of gymnodimine and congeners. PMID- 16268521 TI - Acid-catalyzed ortho-alkylation of anilines with styrenes: an improved route to chiral anilines with bulky substituents. AB - [reaction: see text] Reaction of para-substituted anilines with styrene derivatives at elevated temperatures, when catalyzed by CF3SO3H, results in highly chemoselective ortho-alkylation of the aniline. When R = H, dialkylation can be achieved by varying the ratio of styrene to aniline. Several different substituted anilines and styrenes were examined, and good yields (42-87%) were obtained, except in the case where electron-withdrawing substituents are present on the styrene. PMID- 16268522 TI - Reaction of Huisgen zwitterion with 1,2-benzoquinones and isatins: expeditious synthesis of dihydro-1,2,3-benzoxadiazoles and spirooxadiazolines. AB - [reaction: see text] The zwitterionic intermediate generated from dialkyl azodicarboxylate and triphenylphosphine on reaction with 3-methoxy-1,2 benzoquinones afforded dihydro-1,2,3-benzoxadiazoles. N-Substituted isatins furnished spirooxadiazolines under similar conditions. PMID- 16268523 TI - N-spiro chiral quaternary ammonium bromide catalyzed diastereo- and enantioselective conjugate addition of nitroalkanes to cyclic alpha,beta unsaturated ketones under phase-transfer conditions. AB - [reaction, structure: see text] Conjugate addition of various prochiral nitroalkanes to cyclic alpha,beta-unsaturated ketones was found to be efficiently catalyzed by N-spiro C2-symmetric chiral quaternary ammonium bromide 1b possessing a 3,5-bis(3,4,5-trifluorophenyl)phenyl substituent under solid-liquid phase-transfer conditions to afford the corresponding gamma-nitro ketones in excellent chemical yields with unprecedented levels of diastereo- and enantiocontrol. PMID- 16268524 TI - tert-Butyl (phenylsulfonyl)alkyl-N-hydroxycarbamates: the first class of N-(Boc) nitrone equivalents. AB - [reaction: see text] tert-Butyl (phenylsulfonyl)alkyl-N-hydroxycarbamates 1 have been easily prepared from aldehydes and tert-butyl N-hydroxycarbamate in a methanol-water mixture using sodium benzenesulfinate and formic acid. These sulfones 1 behave as N-(Boc)-protected nitrones 4 in the reaction with organometallics to give N-(Boc)hydroxylamines. Some chemical transformations showing their interest as building blocks in organic synthesis are described. PMID- 16268525 TI - Highly regio- and enantioselective palladium-catalyzed allylic alkylation and amination of dienyl esters with 1,1'-P,N-ferrocene ligands. AB - [reaction, structure: see text] Pd-catalyzed asymmetric allylic alkylation of dienyl acetates 1 and amination of allyl acetates 2 provides the corresponding chiral products in high regio- and enantioselectivities using 1,1'-P,N-ferrocenes L1a and L2d as ligands, respectively. PMID- 16268526 TI - Construction of the cyclovibsanin core via a biogenetically modeled approach. AB - [reaction: see text] Construction of the 15-O-methylcyclovibsanin B core was achieved expediently in eight linear steps utilizing a biogenetically modeled approach. PMID- 16268527 TI - Light-induced coumarin cyclopentannelation. AB - [reaction: see text] Whereas cyclopentenylcarbenes resulting from photocycloaddition of 4-alk-1-ynylcoumarins to 2,3-dimethylbut-2-ene undergo tandem cyclization to hitherto unknown tetracyclic (4 hetera)cyclopent[b,c]acenaphthylenes, the corresponding cyclopentenylnitrenes stemming from 4-cyanocoumarins and the same alkene are converted into tricyclic imines via H-abstraction. PMID- 16268528 TI - Stereocontrolled synthesis of the DE ring system of the marine alkaloid upenamide. AB - [reaction: see text] A stereocontrolled synthesis of the DE fragment (2) of the marine alkaloid upenamide (1) is described. The synthesis proceeds in 12 steps from caprolactone (10) and 20-25% overall yield. PMID- 16268529 TI - Benzylic oxidation catalyzed by dirhodium(II,III) caprolactamate. AB - [reaction: see text] Dirhodium caprolactamate [Rh2(cap)4] is an effective catalyst for benzylic oxidation with tert-butyl hydroperoxide (TBHP) under mild conditions. Sodium bicarbonate is the optimal base additive for substrate conversion. Benzylic carbonyl compounds are readily obtained, and a formal synthesis of palmarumycin CP2 using this methodology is described. PMID- 16268530 TI - Catalytic addition methods for the synthesis of functionalized diazoacetoacetates and application to the construction of highly substituted cyclobutanones. AB - [reaction: see text] Methyl 3-(trialkylsilanyloxy)-2-diazo-3-butenoate undergoes Lewis acid-catalyzed Mukaiyama aldol addition with aromatic and aliphatic aldehydes in the presence of low catalytic amounts of Lewis acids in nearly quantitative yields. Scandium(III) triflate is the preferred catalyst and, notably, addition proceeds without decomposition of the diazo moiety. Diazoacetoacetate products from reactions with aromatic aldehydes undergo rhodium(II)-catalyzed ring closure to cyclobutanones with high diastereocontrol. Examples of complimentary Mannich-type addition reactions with imines are reported. PMID- 16268531 TI - Concise and stereoselective synthesis of the N7-C25 fragment of psymberin. AB - [reaction: see text] The N7-C25 fragment of the potent and selective cytotoxic agent psymberin has been prepared through a short (12 linear steps, 15 total steps) and stereoselective sequence. Highlights of this route include a very rapid construction of the pentasubstituted arene, a substrate-controlled diastereoselective fragment coupling using a Mukaiyama aldol reaction, and an efficient entry into a key tetrahydropyranyl cyanide. PMID- 16268532 TI - Highly efficient synthesis of isoquinolines via nickel-catalyzed annulation of 2 iodobenzaldimines with alkynes: evidence for dual pathways of alkyne insertion. AB - [reaction: see text] A wide range of substituted isoquinolines were synthesized via a highly efficient nickel-catalyzed annulation of the tert-butyl imines of 2 iodobenzaldehydes and various alkynes; examination of the regiochemistry of isoquinolines synthesized indicates that there are two different alkyne insertion pathways for the catalytic reactions. PMID- 16268533 TI - A stereoselective cyclization to carbafuranose derivatives starting from 1,4-bis epoxides. AB - [reactions: see text] A concise synthesis of highly functionalized cyclopentane derivatives via a Brook rearrangement mediated stereoselective linchpin cyclization reaction involving tert-butyldimethylsilyl-1,3-dithianyllithium and homochiral 1,4-bis-epoxides is described. PMID- 16268534 TI - Effective PET and ICT switching of boradiazaindacene emission: a unimolecular, emission-mode, molecular half-subtractor with reconfigurable logic gates. AB - [reaction, structure: see text] We report a unimolecular system functioning as a combinatorial logic circuit for half-subtractor. The emission characteristics can be modulated by chemical inputs, and when followed at two different wavelengths, two functionally integrated logic gates XOR and INHIBIT are obtained. Both logic gates function in the emission mode, and with very large differences in the signal intensity allowing unequivocal assignment of logic-0 and logic-1. PMID- 16268535 TI - Trans- and cis-selective Lewis acid catalyzed hydrogermylation of alkynes. AB - [reactions: see text] The first examples of Lewis acid catalyzed hydrogermylation of alkynes have been demonstrated. It was found that this method has much higher functional group compatibility compared to the known Lewis acid catalyzed hydrosilylation and hydrostannation reactions. Remarkably, the stereochemical outcome of this hydrogermylation reaction depends on the nature of the alkyne used: proceeding via a trans-addition pathway with simple alkynes and cis addition with propiolates. Mechanistic studies strongly support the proposed rationale on the origins of cis-selectivity in the hydrogermylation of activated alkynes. PMID- 16268536 TI - Thermal isomerization of cis,anti,cis-tricyclo[6.3.0.0(2,7)]undec-3-ene to endo tricyclo[5.2.2.0(2,6)]undec-8-ene. AB - [reaction: see text] The gas-phase thermal isomerization of cis,anti,cis tricyclo[6.3.0.0(2,7)]undec-3-ene (1) to endo-tricyclo[5.2.2.0(2,6)]undec-8-ene (2) at 315 degrees C occurs cleanly through a symmetry-forbidden [1,3] suprafacial,retention (sr) pathway. PMID- 16268538 TI - Synthesis of isoxazoles via electrophilic cyclization. AB - [reaction: see text] A variety of 3,5-disubstituted 4-halo(seleno)isoxazoles are readily prepared in good to excellent yields under mild reaction conditions by the reaction of 2-alkyn-1-one O-methyl oximes with ICl, I2, Br2, or PhSeBr. PMID- 16268537 TI - Design, synthesis, and biological evaluation of potent discodermolide fluorescent and photoaffinity molecular probes. AB - [structure: see text] The design, synthesis, and biological evaluation of a series of (+)-discodermolide molecular probes possessing photoaffinity and fluorescent appendages has been achieved. Stereoselective olefin cross-metathesis comprised a key tactic for construction of two of the molecular probes. Three photoaffinity probes were radiolabeled with tritium. PMID- 16268539 TI - Concise synthesis of (+/-)-rhazinilam through direct coupling. AB - [reaction: see text] A concise synthesis of rhazinilam through direct, palladium catalyzed, intramolecular coupling is described. PMID- 16268540 TI - Synthesis and incorporation into DNA of a chemically stable, functional abasic site analogue. AB - [reaction: see text] The abasic site building block 7 for DNA synthesis, containing a methylenephosphinic acid group at C3', was prepared in six steps and was incorporated into DNA via a combination of H-phosphonate and phosphoramidite chemistry. Corresponding oligodeoxynucleotides were shown to be chemically stable under basic conditions and fully functional at the respective hemiacetal center. PMID- 16268541 TI - Electrophilic behavior of the pi delocalized azepinium ion: Friedel-Crafts reactions with benzenes and five-membered aromatic heterocycles. AB - [reaction: see text] Although the reactivity of tropylium ion with aromatic substrates is low, the reaction of azepinium ion with aromatic substrates such as benzene, phenol, furan, and thiophene resulted in the formation of 2-aryl-2H azepine as a major product. An exceptional result in the formation of ring contracted product was observed in the reaction with pyrrole. PMID- 16268542 TI - Synthesis of new artemisinin-derived dimers by self-cross-metathesis reaction. AB - [reaction: see text] New artemisinin-derived dimers, fluorinated or not, have been prepared by a self-cross metathesis reaction in the presence of first- or second-generation ruthenium catalysts without degradation of the endoperoxide bridge and with a good E/Z selectivity (up to 100:0). PMID- 16268543 TI - Cyclohexenones as Michael acceptors in the Staunton-Weinreb annulation: a simple stannane modification for the synthesis of polycyclic systems. AB - [reaction: see text] o-Toluate anions generated via transmetalation from the corresponding tributyl stannane underwent a Michael addition-Dieckmann condensation sequence with various cyclohexenones. This protocol provides an efficient entry into complex polycyclic systems without the use of beta-alkoxy enones hitherto required for the reaction. PMID- 16268544 TI - A six-step asymmetric synthesis of (+)-hyperaspine. AB - [reaction: see text] The ladybird alkaloid, (+)-hyperaspine, has been synthesized in a concise and highly stereoselective manner. The total synthesis was accomplished in six steps and 21% overall yield. PMID- 16268545 TI - Design, synthesis, and biological evaluation of simplified analogues of (+) discodermolide. Additional insights on the importance of the diene, the C7 hydroxyl, and the lactone. AB - [structure: see text] The design, synthesis, and biological evaluation of seven totally synthetic analogues of the antitumor agent (+)-discodermolide are reported. Saturation of the terminal diene system, alteration of the substituents on the lactone, and alkylation of the C7-hydroxyl group reveal significant structure-activity relationships. PMID- 16268546 TI - Uncialamycin, a new enediyne antibiotic. AB - [structure: see text] Laboratory cultures of an undescribed streptomycete obtained from the surface of a British Columbia lichen produce uncialamycin (1), a new enediyne antibiotic. The structure of uncialamycin (1) has been elucidated by analysis of spectroscopic data. Uncialamycin (1) exhibits potent in vitro antibacterial activity against gram-positive and gram-negative human pathogens, including Burkholderia cepacia, a major cause of morbidity and mortality in patients with cystic fibrosis. PMID- 16268547 TI - Mild and reversible dehydration of primary amides with PdCl2 in aqueous acetonitrile. AB - [reaction: see text] A new, mild, and reversible method to convert primary amides to nitriles in good yields using PdCl2 in aqueous acetonitrile is described. PMID- 16268548 TI - Aminoarenethiolate-copper(I)-catalyzed amination of aryl bromides. AB - [reaction, structure: see text] Aminoarenethiolate-copper(I) complexes are known to be efficient catalysts for carbon-carbon bond formation. Here, we show the first examples that these thiolate-copper(I) complexes are efficient for carbon nitrogen bond formation reactions as well. N-Arylation of benzylamine and imidazole with bromobenzene was achieved either in NMP as solvent or under solvent-free conditions in the presence of 2.5 mol % of aminoarenethiolate copper(I) complex only. PMID- 16268550 TI - Copper-catalyzed hydrostannation of activated alkynes. AB - [reactions: see text] [(Ph3P)CuH]6 effectively catalyzes the hydrostannation of activated alkynes with exclusive regioselectivity for alpha-stannation. Syn hydrostannation is observed exclusively for alkynoates. Anti or syn hydrostannation adducts are obtained as products for alkynone substrates. PMID- 16268549 TI - Sequential acid/base-catalyzed polycyclization of tryptamine derivatives. A rapid access to Buchi's ketone. AB - [reaction: see text] The development of an efficient and diastereoselective methodology that allows the rapid construction of the tetracyclic core of the Aspidosperma and Strychnos alkaloid families is described. Our approach relies upon two key steps: a sequential silica gel/potassium tert-butoxide polycyclization of a tryptamine precursor and a tandem oxidative decarboxylation/ring-closing reaction. The assembly of Buchi's ketone, a key intermediate in the synthesis of vindorosine, has been accomplished using this approach. PMID- 16268551 TI - Highly fluorescent oligothiophenes through the incorporation of central dithieno[3,2-b:2',3'-d]pyrrole units. AB - [structures: see text] Dithieno[3,2-b:2',3'-d]pyrrole-based terthiophene (2a-c) and quaterthiophene (3a-c) analogues have been prepared from dithieno[3,2-b:2',3' d]pyrrole (1) via Stille coupling utilizing a one-pot method. In comparison to the parent oligothiophenes (T(n), where n = 2-4), the resulting dithieno[3,2 b:2',3'-d]pyrrole-based systems exhibit enhanced fluorescence efficiencies in solution (up to 53%). These new oligomeric systems also allow the incorporation of solubilizing side chains without the negative steric interactions that typically reduce backbone planarity. PMID- 16268552 TI - Locked pi-expanded chlorins in two steps from simple tetraarylporphyrins. AB - [reaction: see text] Upon tandem Reformatsky reaction, easily accessible porphyrinic ketones give "locked" chlorinic diester. Both ketones and diesters, as bases or palladium complexes, efficiently generate singlet dioxygen, as demonstrated by trapping with cholesterol. PMID- 16268553 TI - Antineoplastic diterpene-benzoate macrolides from the Fijian red alga Callophycus serratus. AB - [structures: see text] Three diterpene-benzoate natural products, with novel carbon skeletons and an unusual proposed biosynthesis, were isolated from extracts of the Fijian red alga Callophycus serratus and identified by a combination of X-ray crystallographic, NMR, and mass spectral analyses. Bromophycolide A (1) displayed cytotoxicity against several human tumor cell lines via specific apoptotic cell death. This represents the first discovery of natural products incorporating a diterpene and benzoate skeleton into a macrolide system. PMID- 16268554 TI - Chirality organization of ferrocenes bearing dipeptide chains of heterochiral sequence. AB - [structures: see text] The symmetrical introduction of two dipeptide chains of heterochiral sequence (-L-Ala-D-Pro-NHPy) into the ferrocene scaffold as a central reverse-turn unit was demonstrated to induce both antiparallel beta-sheet like and type II beta-turn-like structures simultaneously, affording the chirality-organized structure. The ferrocene bearing only one dipeptide chain (-L Ala-D-Pro-NHPy) exhibited a left-handed helically ordered molecular assembly through a network of intermolecular hydrogen bonds instead of intramolecular hydrogen bonds. PMID- 16268555 TI - Systematic assignment of the configuration of flexible natural products by spectroscopic and computational methods: the bistramide C analysis. AB - [reaction: see text] The combination of NMR NOE, chemical shift, and J-coupling measurements with molar rotation and circular dichroism (CD) determinations, including RI-DFT BP86/aug-cc-pVDZ calculations, reduced a candidate pool of 1024 possible stereoisomers of (+)-bistramide C to a single absolute configuration assignment for the 10 stereogenic carbons of the marine natural product. PMID- 16268556 TI - A one-pot regiospecific synthesis of highly functionalized 1,4-benzodioxin derivatives from an electrochemically induced Diels-Alder reaction. AB - [reaction: see text] The anodic oxidation of pyrogallol derivatives produces chemically unstable o-quinone heterodienes, which are trapped in situ by enamine dienophiles through regiospecific inverse-electron-demand Diels-Alder reactions. The possibility of introducing variations in both cycloaddition partners gives rise to highly substituted 1,4-benzodioxin cycloadducts with up to five elements of diversity. The reactions proceed under mild conditions with a good efficiency. The methodology should be amenable to the assembly of libraries of biologically relevant heterocycles. PMID- 16268557 TI - Synthesis and anion binding properties of 2,5-diamidothiophene polypyrrole Schiff base macrocycles. AB - [reaction: see text] Two easy-to-synthesize polypyrrolic 2,5-diamidothiophene Schiff base macrocycles are reported, along with their anion binding properties as determined via UV-vis spectroscopic titrations carried out in dichloroethane. There is a striking difference between the interactions with anions of the two macrocycles, a finding ascribed to differences in their rigidity. For example, the more flexible dipyrromethane-derived macrocycle displays a 1.2:1 hydrogen sulfate versus nitrate selectivity, while its more rigid bipyrrole-derived congener shows a 7.4:1 selectivity in favor to hydrogen sulfate. PMID- 16268558 TI - From ketenimines to ketenes to quinolones: two consecutive pseudopericyclic events. AB - [reaction: see text] N-[2-(Alkyl- or arylthio)carbonyl]phenyl ketenimines undergo cyclization under mild thermal conditions to afford 2-alkyl(aryl)thio-3H-quinolin 4-ones by means of the 1,5-migration of the alkyl(aryl)thio group from the carbonyl carbon to the central carbon atom of the ketenimine fragment followed by the 6pi-electrocyclization of the resulting vinyliminoketene. These 1,5-migration and electrocyclization processes occur via transition states whose pseudopericyclic characteristics have been established on the basis of their magnetic properties, geometries, and NBO analyses. PMID- 16268559 TI - Exploiting the dual reactivity of o-isocyanobenzamide: three-component synthesis of 4-imino-4H-3,1-benzoxazines. AB - [reaction: see text] A multicomponent synthesis of 4-imino-4H-3,1-benzoxazines is developed. Heating a toluene solution of an aldehyde (6), an amine (7), and an isonitrile (5) in the presence of a stoichiometric amount of ammonium chloride at 60 degrees C for 12 h produces the title compound in good to excellent yields. PMID- 16268560 TI - Gold-catalyzed cycloisomerization of o-alkynylbenzaldehydes with a pendant unsaturated bond: [3 + 2] cycloaddition of gold-bound 1,3-dipolar species with dipolarophiles. AB - [reaction: see text] A new and novel Au-catalyzed cycloisomerization of ynals bearing a pendant unsaturated bond leading to synthetically valuable [6.7.n] tricyclic compounds were developed. This study strongly supports the intermediacy of [3 + 2] cycloaddition proposed by DFT calculation and provides an easy access to key skeletons found in a variety of natural products. PMID- 16268561 TI - Asymmetric direct vinylogous Michael reaction of activated alkenes to nitroolefins catalyzed by modified cinchona alkaloids. AB - [reaction: see text] The first organocatalytic and asymmetric direct vinylogous Michael reaction that employs the electron-deficient vinyl malononitriles as the nucleophilic species has been reported. The novel transformations exhibit exclusive gamma-selectivity and high diastereo- and enantioselectivity in the addition to nitroolefins, which give the multifunctional products with two vicinal chiral centers. PMID- 16268562 TI - Taiwankadsurins A, B, and C, three new C19 homolignans from Kadsura philippinensis. AB - [structures: see text] Three novel C19 homolignans, designated taiwankadsurins A (1), B (2), and C (3), were isolated from the aerial parts of Taiwanese medicinal plant Kadsura philippinensis. The structures of 1-3, which have a 3,4-{1'-[(Z) 2''-methoxy-2''-oxo-ethylidene]}-pentano(2,3-dihydro-benzo[b]furano)-3-(2''' methoxycarbonyl-2'''-hydroxy-2''',3'-epoxide) skeleton, were determined by spectroscopic analyses, especially 2D NMR techniques (HMBC and NOESY). Compound 2 exhibited mild cytotoxicity against human KB and Hela tumor cells. PMID- 16268563 TI - General synthesis of thiophene and selenophene-based heteroacenes. AB - [reaction: see text] A new intramolecular triple cyclization of bis(o haloaryl)diacetylenes, via dilithiation followed by reaction with chalcogen elements, produces pi-conjugated compounds containing heterole-1,2-dichalcogenin heterole fused tricyclic skeletons. The subsequent dechalcogenation with copper metal affords a series of thiophene- and selenophene-based heteroacenes. PMID- 16268565 TI - Palladium-catalyzed enantioselective conjugate addition of arylboronic acids. AB - [reaction: see text] The first asymmetric palladium-catalyzed conjugate addition of arylboronic acids to alpha,beta-unsaturated aldehydes, ketones, and esters is described. For cyclic substrates, excellent chemo-, regio-, and enantioselectivities are achieved when a Pd(O2CCF3)2/DuPHOS catalyst is applied. PMID- 16268564 TI - Highly enantioselective synthesis of rigid, quaternary 1,4-benzodiazepine-2,5 diones derived from proline. AB - [reaction: see text] Proline-derived 1,4-benzodiazepine-2,5-diones are extremely useful scaffolds in medicinal chemistry. In this paper, we describe a protocol for retentive C3 alkylation of these materials, thus accomplishing the direct synthesis of enantiopure quaternary 1,4-benzodiazepine-2,5-diones. The high enantioselectivities (up to 99.5%) are attributed to memory of chirality. PMID- 16268566 TI - New tandem Zn-promoted Brook rearrangement/ene-allene carbocyclization reactions. AB - [reaction: see text] A new tandem reaction was developed for the carbocyclization reaction of propargylic zinc reagents. First, we have shown that zinc salt promotes the Brook rearrangement into the carbanionic species followed by a stereospecific Zn-ene-allene carbocyclization reaction to lead to the corresponding cyclopentylmethylzinc derivatives. A single diastereoisomer is formed, even when a tertiary and a quaternary center are linked in the process. PMID- 16268567 TI - Asymmetric synthesis of 2-aryl-2,3-dihydro-4-quinolones by rhodium-catalyzed 1,4 addition of arylzinc reagents in the presence of chlorotrimethylsilane. AB - [reaction: see text] The first catalytic asymmetric synthesis of 2-aryl-2,3 dihydro-4-quinolones has been developed by way of a rhodium-catalyzed 1,4 addition of arylzinc reagents to 4-quinolones. These 1,4-adducts can be obtained with high enantioselectivity by the use of (R)-binap as a ligand, and high yields are realized by conducting the reactions in the presence of chlorotrimethylsilane. PMID- 16268568 TI - C2-symmetric bisprolinamide as a highly efficient catalyst for direct aldol reaction. AB - [reaction: see text] The catalytic activity of the prolinamide-type catalysts may be improved by introducing additional prolinamide moiety into the catalyst, while the enantioselectivity can still be maintained or further improved. A C2 symmetric bisprolinamide with two prolinamide moieties has been found to be an excellent catalyst for direct aldol reaction with more than doubled reactivity and better asymmetric induction than its monoprolinamide counterpart. PMID- 16268569 TI - A convergent total synthesis of ustiloxin D via an unprecedented copper-catalyzed ethynyl aziridine ring-opening by phenol derivatives. AB - [reaction: see text] The ustiloxins are a family of heterodetic cyclopeptides that have been isolated from the water extracts of false smut balls on the panicles of rice plants caused by the fungus Ustilaginoidea virens. A concise total synthesis of ustiloxin D has been achieved via an unprecedented ethynyl aziridine ring-opening of phenol derivatives. The longest linear sequence of the synthesis is 15 steps from commercially available compounds. PMID- 16268570 TI - ZnCl2-mediated stereoselective addition of terminal alkynes to D-(+) mannofuranosyl nitrones. AB - [reaction: see text] An optimized process for the addition of terminal alkynes to chiral nitrones using ZnCl2 and NEt3 in toluene is reported. The new reaction protocol is facile to perform and cost-effective. The resulting optically active propargyl N-hydroxylamines are isolated in good to excellent yield and high diastereoselectivity. PMID- 16268571 TI - Synthesis of the C6-C21 segment of amphidinolide E. AB - [reaction: see text] A convergent approach to a C6-C21 segment of the polyketide amphidinolide E has been developed through combination of three subunits by allenylindium bromide-aldehyde addition and Suzuki sp2-sp3 coupling. PMID- 16268572 TI - N-trialkylsilyl bistrifluoromethanesulfonimides (R3SiNTf2) are powerful catalysts for the highly efficient alpha-amido alkylation reactions of silicon-based nucleophiles. AB - [reaction: see text] In situ formed N-trialkylsilyl bistrifluoromethanesulfonimides (R3SiNTf2) species have been shown to efficiently catalyze the nucleophilic substitution reactions of chiral 5-oxypyrrolidin-2-ones by silicon-based nucleophiles. The reaction rates were significantly accelerated in comparison to the cases where the usual triflate catalysts are used. Adducts were obtained in high yields and usual stereoselectivities within short reaction times, and the process was compatible with a semipreparative scale. PMID- 16268573 TI - syn-selective direct catalytic asymmetric Mannich-type reactions of hydroxyketones using Y{N(SiMe3)2}3/linked-BINOL complexes. AB - [reaction, structure: see text] Chiral Y{N(SiMe3)2}3/linked-BINOL catalyst generated Y-enolate in situ from various hydroxyketones (R2 = aryl, heteroaryl). Beta-amino-alpha-hydroxy ketones (R1 = aryl, heteroaryl, alkenyl) were obtained syn-selectively (up to 96/4) in high ee (up to 98%) and good yield (up to 98% yield). PMID- 16268574 TI - Efficient rhodium-catalyzed asymmetric hydrogenation for the synthesis of a new class of N-aryl beta-amino acid derivatives. AB - [reaction: see text] N-Aryl beta-amino esters were obtained by asymmetric hydrogenation of a new class of N-aryl beta-enamino esters. High conversions and up to 96.3% ee values were achieved with a Rh-TangPhos catalyst. PMID- 16268578 TI - DNA cleavage by the photolysis of cyclopentadienyl metal complexes: mechanistic studies and sequence selectivity of strand scission by CpW(CO)3CH3. AB - [Reaction: see text]. The photolysis of CpW(CO)3Me has been shown to produce methyl radicals and to cleave DNA in a single-stranded manner, and preliminary evidence implicated a carbon-centered radical in this process. In this work, the mechanism of strand scission in this reaction was determined to occur by hydrogen atom abstraction from the 4'- and 5'-positions of the deoxyribose moiety of the backbone of DNA. Additionally, in a side reaction that does not lead to frank strand scission, all four bases of DNA are methylated under these conditions; however, none of these base or backbone modifications lead to the formation of abasic sites. PMID- 16268579 TI - Determination of the absolute configuration of three as-hydrindacene compounds by vibrational circular dichroism. AB - [Structure: see text]. The absolute configurations of three compounds with a rigid 1,8-disubstituted as-hydrindacene skeleton have been determined using vibrational circular dichroism spectroscopy and quantum chemical calculations. Experimental spectra were compared to B3LYP/6-31G and B3LYP/cc-pVTZ level predicted spectra. Based on the agreement between the predicted and experimental spectra, the stereochemistry could be assigned with high confidence. The results were found to be in agreement with ECD determinations and/or predictions based on the applied asymmetric methods in the synthetic route. PMID- 16268580 TI - Alpha,beta-unsaturated ketoximes carrying a terminal pyridine or quinoline subunit as building blocks for supramolecular syntheses. AB - [Structure: see text]. The crystal structures of a new series of alpha,beta unsaturated ketoximes, 8-14, carrying the terminal 4-pyridinyl, 3-pyridinyl, or 4 quinolinyl subunit have been investigated by X-ray structural analysis. The dominating intermolecular interaction in all structures, except 11, is the head tail OH...N hydrogen bond between the oxime moiety and the nitrogen atom of the heterocyclic unit. This intermolecular interaction generates infinite chains, which are cross-linked by CH...O/N/Cl or CH...pi interactions. Compound 10 has been shown to adopt a double-helical structure in the crystalline state. Compound 11 represents the only case where the unexpected head-head NOH...N(OH) hydrogen bonds determine the crystal packing. Both hydrogen-bonding and aromatic interactions stabilize the crystal structures of 8-14. PMID- 16268581 TI - O-confused oxaporphyrin--an intermediate in formation of 3-substituted 2-oxa-21 carbaporphyrins. AB - [Reaction: see text]. A condensation of 2,4-bis(phenylhydroxymethyl)furan with pyrrole and p-toluylaldehyde in the presence of ethanol formed 5,20-diphenyl 10,15-di(p-tolyl)-2-oxa-3-ethoxy-3-hydro-21-carbaporphyrin [(H,EtO)OCPH]H2. The new carbaporphyrinoid has 1H NMR features of an aromatic molecule, including the upfield shift of the inner H(21) atom (-5.48 ppm). Addition of acid removes the ethoxy substituent and converts [(H,EtO)OCPH]H2 into the dication of "true" O confused oxaporphyrin {[(H)OCPH]H3}2+ via an exocyclic C(3)-O bond cleavage followed by an elimination of the ethoxy group as determined by 1H NMR. Addition of ethanol, water, or pyrrole converts {[(H)OCPH]H3]2+ into [(H,EtO)OCPH]H2, [(H,OH)OCPH]H2, or pyrrole appended O-confused porphyrin [(H,pyrrole)OCPH]H2, respectively. The reaction of [(H,OEt)OCPH]H2 with silver(I) acetate yields a stable Ag(III) complex [(H,OEt)OCP]AgIII substituted at the C(3) position by the ethoxy group and hydrogen. Coordination through the nitrogen donors is reflected by the presence of 107/109Ag scalar splitting seen for the selected -H pyrrolic signals. Addition of TFA to [(H,OEt)OCP]AgIII produces a weakly aromatic O confused porphyrin complex {[(H)OCP]AgIII}+. In the course of this reversible process the tetrahedral-trigonal rearrangements originate at the C(3) atom but extend its consequences on the whole structure. Insertion of silver into the hydroxy analogue of [(H,OEt)OCPH]H2, i.e., [(H,OH)OCPH]H2, was accompanied by ligand oxidation, yielding carbaporpholactone which contains the lactone functionality instead of the regular furan moiety embedded in the carbaporphyrin ligand of [(O)OCP]AgIII. The structure was determined by X-ray crystallography. The macrocycle is only slightly distorted from planarity, and silver(III) is located in the NNNC plane. PMID- 16268582 TI - Internally solvated cyclopentadienyllithium compounds: structural integrity of the Cp-Li+ moiety. NMR, dynamics, X-ray crystallography, and calculations. AB - [Structure: see text]. N(CH2CH2OCH3)2 are as follows: T = CHCH(CH3)2, 6; T = (CH2)2, 10; T = (CH2)3, 14. The results of NOE NMR experiments for 6, 10, and 14 together with X-ray crystallography of 14 support internally coordinated monomeric structures for all three compounds. Models have been constructed for 6, 10, and 14 from modifications of an internally solvated allylic lithium compound at the B3LYP level of theory using basis set 6-311G*. The resulting structural features are very similar to those obtained from the NMR and crystallographic data. In addition, 13C NMR shifts obtained with the GIAO procedure using the results of the B3LYP/6-311G* calculations are closely similar to the experimental shifts, which validate B3LYP as a suitable model for these compounds. The Li+ centroid distance of ca. 1.9 A to 2.0 A obtained for 6, 10, and 14 is common to most crystallographic data for externally solvated Cp-Li+ compounds as well as one which incorporates a (CpLiCp)- triple ion. It is concluded that the ligand tether and the stereochemistry around Li+ accommodate to maintain the structural integrity of Cp-Li+. NMR and crystallography show 14 to be chiral. Carbon-13 NMR line shape changes are attributed to inversion via a lateral wobble mechanism with DeltaH++ = 6 kcal x mol(-1) and DeltaS++ = -2 eu. It is also shown that a 6,6-dimethylfulvene is deprotonated at methyl by LiN(CH2CH2OCH3)3 as well as by butyllithium in the presence of PMDTA producing isopropenyl Cp-Li+ compounds 24 and 25, respectively. NMR line shape changes of the sample containing 24 have been qualitatively interpreted to result from a combination of fast transfer of coordinated ligand between faces of the carbanion plane as well as a lithium exchange process. PMID- 16268583 TI - Probing the intermediates of halogen addition to alkynes: bridged halonium versus open halovinyl cation; a theoretical study. AB - [Reaction: see text]. Intermediates formed in halogen addition (X = Br, Cl, F) to alkynes (ethyne, propyne, 2-butyne, trifluoromethylethyne, trimethylsilylethyne, and 1-trimethylsilylpropyne) were studied computationally by MP2 at the MP2/6 311++G(3df,3pd) level and/or by DFT at the B3LYP/6-31+G(d) level. Structure optimization and frequency calculations were performed to identify the minima and to obtain their relative energies. PCM calculations (with H2O, CH2Cl2, and CCl4 as model solvents) were employed to examine solvation effects on the relative stabilities in the resulting bridged halonium, -halovinyl, or -halovinyl cations. GIAO-MP2 and GIAO-DFT calculations were employed to compute NMR chemical shifts (13C, 19F, and 29Si as appropriate). In selected cases, PCM-GIAO calculations were also performed to investigate the extent of solvent effects on the computed NMR shifts. The NPA-derived charges and the GIAO shifts were examined in comparative cases to shed light on structural features. In several cases, structure optimization starting with the -halovinyl cations resulted in halovinyl cations (via formal hydride shift or trimethylsilyl shift). With the CF3 derivative (when X = F), a formal F shift results in polyfluoroallyl cation generation from fluorovinyl cation as starting geometry. PMID- 16268584 TI - Synthesis of 2H-1,2-benzothiazine 1,1-dioxides via heteroannulation reactions of 2-iodobenzenesulfonamide with ketone enolates under S(RN)1 conditions. AB - [Reaction: see text]. 2-iodobenzenesulfonamide (1a) underwent photostimulated S(RN)1 reactions in liquid NH3 with the potassium enolates derived from acetone, pinacolone, butanone, and 3-methyl-2-butanone to give fair to good yields of 2H 1,2-benzothiazine 1,1-dioxides 2. Reductive dehalogenation of 1a was found to predominate in photoinduced reactions of 1a with 3-pentanone, 2-methyl-3 pentanone, and 2,4-dimethyl-3-pentanone, the extent of reduction being proportional to the number of beta-hydrogen atoms present in the ketone enolate. Isotopic labeling studies with 2,4-dimethyl-3-pentanone-d14 (24) confirmed the major role of the beta-hydrogens in the reduction process. Reactions of 1a with cyclopentanone, cyclohexanone, and cyclooctanone enolates afforded new tricyclic benzothiazine derivatives 26-29. Attempts to extend the heteroannulation reaction to the preparation of 2H-1,2-benzothiazin-3(4H)-one 1,1-dioxides 3 (R = H, Ph) through reactions of 1a with tert-butyl acetate and ethyl phenylacetate enolates resulted only in hydrodehalogenation of 1a. However, oxazoline anion 30, a synthetic equivalent of ethyl phenylacetate, was successfully employed in an alternative S(RN)1-based synthesis of benzothiazine 3 (R = Ph). PMID- 16268585 TI - Dual behavior of masked o-benzoquinones in intramolecular Diels-Alder reactions. Expedient synthesis of highly functionalized cis-decalins from 2-methoxyphenols. AB - [Reaction: see text]. The potential dual behavior as dienes and dienophiles of the diene moieties of masked o-benzoquinones (MOBs) 10a-e-12a-e, generated upon oxidation of 2-methoxyphenols 1-3 with BTIB in the presence of appropriate dienols, in their intramolecular Diels-Alder (IMDA) reactions has been examined. The IMDA reactions of MOBs 10a-d, 11a,b,d, and 12a,b,d resulted in highly functionalized oxatricyclic compounds 18a-d, 19a,b,d, and 20a,b,d, respectively, with concomitant formation of cis-decalin derivatives 21a-d, 22a,b,d, and 23a,b,d in a highly regio- and stereoselective manner. However, the MOBs 10e-12e provided exclusively oxatricyclic compounds 18e-20e. The formation of cis-decalins in these IMDA reactions illustrates the dienophilic character of MOBs, in addition to their behavior as dienes. The ratio of the two cycloadducts obtained in each reaction as a result of the dual character of MOBs depends on the nature and/or position of the substituents on both the cyclohexadienone moiety and the added 2,4-dienol. The majority of the cycloadducts resulted from the diene property of MOBs in intramolecular Diels-Alder reactions smoothly underwent Cope rearrangement to furnish cis-decalins as sole products in excellent to quantitative yields that provides a short and efficient entry to polyfunctionized cis-decalins from 2-methoxyphenols. Furthermore, the variation of dienophilic and diene characters of MOBs in the IMDA reactions with the electron-donating or electron-withdrawing substituent of both cyclohexadienone moiety and the added conjugated acyclic diene or 2,4-dienol has been studied in detail. PMID- 16268587 TI - A DFT investigation of alkyne bromination reactions. AB - [Structure: see text]. A DFT calculation study of the addition reaction between molecular bromine and the number of symmetrical or unsymmetrical substituted alkynes 1 (R-CC-R'), where R = R' = H (1a), Me (1b), t-Bu (1c), or Ph (1d), or R = H and R' = Me (1e), t-Bu (1f), or Ph (1g), was performed. Two possible reactions were checked: (a) the reactions suitable for the gas-phase interactions, which start from a 1:1 Br2-alkyne pi-complex and do not enter into a 2:1 Br2-alkyne pi-complex; and (b) the processes passing through a 2:1 Br2 alkyne pi-complex, which look more realistic for the reactions in solutions. The structures of the starting reactants and the final products as well as the possible stable intermediates have been optimized. The transition states of the predicted process have been found. Both trans- and cis-dibromoalkenes (2 and 3) may ensue without the formation of ionic intermediates from a pi-complex of two bromine molecules with the alkyne (solution reactions). The geometry around the double bond forming in dibromoalkenes strongly depends on the nature of the substituents at the triple bond. The "cluster model" was also used for the prediction of solvent influence on the value of the activation barrier of the but 2-yne (1b) bromination reaction. PMID- 16268586 TI - Biosynthesis of isoprenoids. purification and properties of IspG protein from Escherichia coli. AB - [Structure: see text]. The IspG protein is known to catalyze the transformation of 2-C-methyl-d-erythritol 2,4-cyclodiphosphate into 1-hydroxy-2-methyl-2-(E) butenyl 4-diphosphate in the nonmevalonate pathway of isoprenoid biosynthesis. We have found that the apparent IspG activity in the cell extracts of recombinant Escherichia coli cells as observed by a radiochemical assay can be enhanced severalfold by coexpression of the isc operon which is involved in the assembly of iron-sulfur clusters. The recombinant protein was isolated by affinity chromatography under anaerobic conditions. With a mixture of flavodoxin, flavodoxin reductase, and NADPH as the reducing agent, stringent assay methods based on photometry or on 13C NMR detection of multiply 13C-labeled substrate/product ratios afforded catalytic activities greater than 60 nmol mg( 1) min(-1) for the protein "as isolated" (i.e., without reconstitution of any kind). Lower apparent activities were found using photoreduced deazaflavin as an artifactual electron donor, whereas dithionite was unable to serve as an artificial electron donor. The apparent Michaelis constant for 2-C-methyl-D erythritol 2,4-cyclodiphosphate was 700 microM. The enzyme was inactivated by EDTA and could be reactivated by Mn2+. The pH optimum was at 9.0. The protein contained 2.4 iron ions and 4.4 sulfide ions per subunit. The replacement of any of the three conserved cysteine residues afforded mutant proteins which were devoid of catalytic activity and contained less than 6% of Fe2+ and less than 23% of S2- as compared to the wild-type protein. Sequence comparison indicates that putative IspG proteins of plants, the apicomplexan protozoan Plasmodium falciparum, and bacteria from the Bacteroidetes/Chlorobi group contain an insert of about 170-320 amino acid residues as compared with eubacterial enzymes. PMID- 16268588 TI - Ag NMR as a tool for mechanistic studies of Ag-catalyzed reactions: evidence for in situ formation of alkyn-1-yl silver from alkynes and silver salts. AB - [Structure: see formula]. As demonstrated by 1H, 13C, and 109Ag NMR, a pi-alkyne Ag complex and then an alkynyl silver are in situ formed from alkyne and silver salt in conditions related to those used for Ag-catalyzed alkynylation or for Ag/Pd-catalyzed sp-sp2 cross-coupling reactions. This finding allows for a rationale of the mechanisms of these reactions. PMID- 16268589 TI - N-sulfonylbenzotriazoles as advantageous reagents for C-sulfonylation. AB - [Structure: see text]. Reactions of readily available N-(alkyl-, aryl-, and heteroarylsulfonyl)benzotriazoles 3a-h with diverse nitriles, reactive heteroaromatics, alkylheteroaromatics, sulfones, and esters produced alpha cyanoalkyl sulfones 5a-i, sulfonylheteroaromatics 7a-e, alpha (sulfonylalkyl)heterocycles 9a-f, alpha-sulfonylalkyl sulfones 11a-g, and esters of alpha-sulfonyl acids 14a-c, respectively, in synthetically useful to excellent yields. The results represent the first examples of the successful application of sulfonylazoles for C-sulfonylation. PMID- 16268591 TI - Palladium-catalyzed cross-coupling of Baylis-Hillman acetate adducts with organosilanes. AB - [Reaction: see text]. A cross-coupling reaction between acetates of Baylis Hillman adducts and organosilanes is described. A nonconventional solvent poly(ethylene glycol) (PEG) is used as the reaction medium. PMID- 16268590 TI - Universal solid supports for the synthesis of oligonucleotides via a transesterification of H-phosphonate diester linkage. AB - [Structure: see text]. Three universal solid supports exhibiting an hydroxyl function were prepared. The introduction of a first H-phosphonate diester linkage which was kept throughout the elongation allowed the release of 3'-hydroxyl oligonucleotides by a transesterification mechanism. The transesterification was performed in a few minutes with either amino alcohols or K2CO3/methanol. Starting from a hydroxyl solid support, tandem oligonucleotides were synthesized and the solid support was easily recyclable. This strategy was extended to the release of an oligonucleotide from the solid support by a nonbasic treatment opening the way to the synthesis of base-sensitive oligonucleotides thanks to the selective deprotection of a hydroxyl in beta of the H-phosphonate diester linkage. PMID- 16268592 TI - Alpha-nitro ketone synthesis using N-acylbenzotriazoles. AB - [Reaction: see text]. Readily available N-acylbenzotriazoles 2a-l (derived from a variety of aliphatic, (hetero)aromatic, and N-protected alpha-amino carboxylic acids) smoothly convert primary 3a-c and alpha-functionalized primary nitroalkanes 3d into the corresponding alpha-nitro ketones 5a-p in yields of 39 86% (average 63%). PMID- 16268593 TI - Employment of palladium pincer-complexes in phenylselenylation of organohalides. AB - [Reaction: see text]. Palladium pincer-complex-catalyzed selenylation of propargyl-, allyl-, benzyl-, and benzoyl halides could be achieved under mild reaction conditions employing trimethylstannylphenylselenide as selenylating agent. This reaction has a high functional group tolerance as carbmethoxy, tosylamino, nitro, aryl bromide, and unprotected hydroxy groups are tolerated. Mechanistic studies indicate that the catalytic cycle is initiated by formation of a selenium-coordinated palladium pincer-complex, which subsequently reacts with the electrophilic substrate. DFT calculations were performed to explore the mechanism of the transfer of the organoselenium group from palladium to the substrate. These modeling studies revealed some interesting differences between the mechanism of the organoselenium group transfer and (the previously published) organostannane transfer reactions. PMID- 16268595 TI - Aromatic derivatives and tellurium analogues of cyclic seleninate esters and spirodioxyselenuranes that act as glutathione peroxidase mimetics. AB - [Reaction: see text]. Several novel organoselenium and tellurium compounds were prepared and evaluated as mimetics of the selenoenzyme glutathione peroxidase, which protects cells from oxidative stress by reducing harmful peroxides with the thiol glutathione. The compounds were tested for catalytic activity in a model system wherein tert-butyl hydroperoxide or hydrogen peroxide were reduced with benzyl thiol and the rate of the reaction was measured by monitoring the formation of dibenzyl disulfide. Thus, aromatic derivatives 19, 22, 24, and 25 proved to be inferior catalysts compared to the parent cyclic seleninate ester 14 and spirodioxyselenurane 16. In the case of 19 and 22, this was the result of their rapid conversion to the relatively inert selenenyl sulfides 31 and 32, respectively. In general, hydrogen peroxide was reduced faster than tert-butyl hydroperoxide in the presence of the selenium-based catalysts. The cyclic tellurinate ester 27 and spirodioxytellurane 29 proved to be superior catalysts to their selenium analogues 14 and 16, respectively, resulting in the fastest reaction rates by far of all of the compounds we have investigated to date. Oxidation of 29 with hydrogen peroxide produced the unusual and unexpected peroxide 33, in which two hypervalent octahedral tellurium moieties are joined by ether and peroxide bridges. The structure of 33 was confirmed by X-ray crystallography. Although 33 displayed strong catalytic activity when tested independently in the model system, its relatively slow formation from the oxidation of 29 rules out its intermediacy in the catalytic cycle of 29. PMID- 16268594 TI - Practical synthesis of a neuropeptide Y antagonist via stereoselective addition to a ketene. AB - [Reaction: see text]. The synthesis of neuropeptide Y antagonist 1, currently under clinical investigation for the treatment of obesity, is described. The convergent synthesis from trans-spirolactone carboxylic acid intermediate 2a and aminopyrazole 3 is predicated on a stereoselective route to the former. The coupling reaction of ethyl 4-oxocyclohexanecarboxylate (10a) with lithiated isonicotinamide 11 was investigated in detail, but even optimized conditions only provided a 45:55 ratio of trans:cis isomers (12a:12b). While selective crystallization schemes were developed to isolate the thermodynamically less stable trans isomer 2a, improved stereocontrol was subsequentially achieved by the application of ketene chemistry. The ketene formation and quench was investigated under a variety of conditions aimed at maximizing the trans:cis ratio. Reacting a mixture of carboxylic acids 2a and 2b with POCl3 in THF, followed by concomitant addition of tert-butyl alcohol in the presence of TMEDA at 35 degrees C provided a 4:1 ratio of trans:cis tert-butyl esters (18a:18b) via in situ ketene formation. Ester hydrolysis, followed by selective crystallization of undesired 2b as the HCl salt, led to isolation of 2a in 47% overall yield. Aminopyrazole intermediate 3 was synthesized via the condensation reaction of 2 fluorophenylhydrazine hydrochloride (4a) with acrylonitrile derivative 5 in 65 70% yield. Coupling of advanced intermediates 2a and 3b via activation with thionyl chloride gave a 92% yield of 1. PMID- 16268596 TI - o-hydroxylmethylphenylchalcogens: synthesis, intramolecular nonbonded chalcogen...OH interactions, and glutathione peroxidase-like activity. AB - [Structure: see text]. The synthesis and characterization of a series of organochalcogen (Se, Te) compounds derived from benzyl alcohol 13 are described. The synthesis of the key precursor dichalcogenides 15, 22, and 29 was achieved by the ortho-lithiation route. Selenide 18 was obtained by the reaction of the dilithiated derivative 14 with Se(dtc)2. Oxidation of 15 and 22 with H2O2 afforded the corresponding cyclic ester derivatives 17 and 24, respectively. Oxidation of selenide 18 with H2O2 affords the spirocyclic compound 19. The presence of intramolecular interactions in dichalcogenides 15 and 22 has been proven by single-crystal X-ray studies. The cyclic compounds 17 and 19 have also been characterized by single-crystal X-ray studies. GP(X)-like antioxidant activity of selenium compounds has been evaluated by the coupled bioassay method. Density functional theory calculations at the mPW1PW91 level on ditelluride 22 have identified a fairly strong nonbonding interaction between the hydroxy oxygen and tellurium atom. The second-order perturbation energy obtained through NBO analysis conveys the involvement of n(O) --> sigma(Te-Te) orbital overlap in nonbonding interaction. Post wave function analysis with the Atoms in Molecules (AIM) method identified distinct bond critical point in 15 and 22 and also indicated that the nonbonding interaction is predominantly covalent. Comparison between diselenide 15 and ditelluride 22 using the extent of orbital interaction as well as the value of electron density at the bond critical points unequivocally established that a ditelluride could be a better acceptor in nonbonding interaction, when the hydroxy group acts as the donor. PMID- 16268597 TI - Deprotonation of calixarenes in acetonitrile. AB - [Structure: see text]. The pKa values for calixarenes in MeCN have been determined by selective titration with bases using a spectroscopic method. These values are as follows: calix[4]arene pKa(1) = 19.06 +/- 0.22, pKa(2) > 33; calix[6]arene pKa(1) = 15.59 +/- 0.06, pKa(2) = 23.85 +/- 0.35, pKa(3) > 33; calix[8]arene pKa(1) = 17.20 +/- 0.20, pKa(2) = 20.32 +/- 0.31, pKa(3) > 33. The trends in acidity are rationalized using structures generated by a DFT model. For mono-deprotonation, the degree and nature of hydrogen bonding in the anion is the dominant factor; for di-deprotonation, spatial separation of the anionic charges becomes important. PMID- 16268598 TI - Concise and practical synthesis of C-glycosyl ketones from sugar benzothiazoles and their transformation into chiral tertiary alcohols. AB - [Reaction: see text]. A collection of 13 unsymmetrical ketones, each one featuring a sugar (d-glucosyl, d-galactosyl, d-mannosyl, and l-fucosyl) and an aglycone moiety (phenyl, 2-thiazolyl, TMS-ethynyl, allyl, and 1-propenyl) was prepared by a uniform route based on the use of benzothiazole as a carbonyl group equivalent. Succinctly, C-glycosylbenzothiazoles readily prepared by addition of 2-lithiobenzothiazole to sugar lactones and deoxygenation, were subjected to a one-pot reaction sequence involving N-methylation of the heterocyclic ring by MeOTf, treatment of the N-methylbenzothiazolium salt with a Grignard reagent, and HgCl(2)-promoted hydrolysis of the benzothiazoline thus formed. The resulting ketones were isolated in yields varying from 35 to 80%. Treatment of the sugar ketones with various organometals containing the phenyl, 2-thiazolyl, TMS ethynyl, or ethynyl group as a substituent afforded chiral tertiary alcohols. These addition reactions were highly stereoselective as observed by crude NMR analysis and isolation of a single epimer in high yield in each case examined. However, because of the complexity of the reagents involved, the stereochemical outcome of these reactions appears to be difficult to rationalize by simple classical steric models, thus, ab initio studies taking into account the role of the sugar fragment are advisable. An interesting synthetic elaboration of a propargylic alcohol containing the thiazole ring into a propargylic alcohol bearing the formyl and carboxylate groups is reported. PMID- 16268599 TI - Highly blue luminescent triazine-amine conjugated oligomers. AB - [Reaction: see text]. The novel synthetic strategy and optical properties of highly fluorescent, triazine-amine conjugated oligomers are described herein. Under basic conditions, aromatic dinitrile compounds, NC-C6H4-X-C6H4-CN (X = NMe, O, CH2), underwent cyclic trimerization of the cyano groups at both ends to give a series of triazine-containing oligomers. The oligomers can be expressed as (2n + 1) mer, where n represents the number of triazine rings in the oligomer. The absorption maximum of an amine-conjugated trimer (X = NMe, n = 1) was outstandingly red-shifted as compared with those of the other trimers (X = CH2, O). In acidic media, the amine-conjugated trimer showed two-step bathochromic shifts caused by protonation. The absorption maxima of the amine-conjugated (2n + 1) mers (X = NMe, n = 1-4) did not depend on n; instead, shoulder peaks appeared in the long-wavelength region when n 2. The oligomers involving alternate conjugation of triazines and NMe groups through phenylene groups showed strong fluorescence in chloroform. In particular, the pentamer was the most efficient blue emitter (PhiF = 0.82). The other triazine-containing oligomers (X = CH2, O) did not show fluorescence at all. Therefore, it is concluded that the emission properties are due to the strong electron-donating and accepting abilities of the NMe and triazine moieties, respectively. PMID- 16268600 TI - Modeling SN2 reactions in methanol solution by ab initio calculation of nucleophile solvent-substrate clusters. AB - [Structure: see text]. Ab initio calculations were used to study the S(N)2 reactions of the CH3OCH2I molecule with a methoxide ion (CH3O-) and a methanol molecule by systematically building up the reaction system with explicit incorporation of the methanol solvent molecules. For the reaction of CH3OCH2I with a methoxide ion, the explicit incorporation of the methanol molecules to better solvate the methoxide ion led to an increase in the barrier to reaction. For the reaction of CH3OCH2I with a methanol molecule, the explicit incorporation of the methanol molecules led to a decrease in the barrier to reaction because of an inclination of this reaction to proceed with the nucleophilic displacements accompanied by proton transfer through the H-bonding chain. The H-bonding chain served as both acid and base catalysts for the displacement reaction. A ca. 10(15)-fold acceleration of the methanol tetramer incorporated S(N)2 reaction was predicted relative to the corresponding methanol monomer reaction. The properties of the reactions examined are discussed briefly. PMID- 16268602 TI - Acid-, base-, and lewis-acid-catalyzed heterolysis of methoxide from an alpha hydroxy-beta-methoxy radical: models for reactions catalyzed by coenzyme B12 dependent diol dehydratase. AB - [Reaction: see text].A model for glycol radicals was employed in laser flash photolysis kinetic studies of catalysis of the fragmentation of a methoxy group adjacent to an alpha-hydroxy radical center. Photolysis of a phenylselenylmethylcyclopropane precursor gave a cyclopropylcarbinyl radical that rapidly ring opened to the target alpha-hydroxy-beta-methoxy radical (3). Heterolysis of the methoxy group in 3 gave an enolyl radical (4a) or an enol ether radical cation (4b), depending upon pH. Radicals 4 contain a 2,2 diphenylcyclopropane reporter group, and they rapidly opened to give UV observable diphenylalkyl radicals as the final products. No heterolysis was observed for radical 3 under neutral conditions. In basic aqueous acetonitrile solutions, specific base catalysis of the heterolysis was observed; the pK(a) of radical 3 was determined to be 12.5 from kinetic titration plots, and the ketyl radical formed by deprotonation of 3 eliminated methoxide with a rate constant of 5 x 10(7) s(-1). In the presence of carboxylic acids in acetonitrile solutions, radical 3 eliminated methanol in a general acid-catalyzed reaction, and rate constants for protonation of the methoxy group in 3 by several acids were measured. Radical 3 also reacted by fragmentation of methoxide in Lewis-acid catalyzed heterolysis reactions; ZnBr2, Sc(OTf)3, and BF3 were found to be efficient catalysts. Catalytic rate constants for the heterolysis reactions were in the range of 3 x 10(4) to 2 x 10(6) s(-1). The Lewis-acid-catalyzed heterolysis reactions are fast enough for kinetic competence in coenzyme B12 dependent enzyme-catalyzed reactions of glycols, and Lewis-acid-catalyzed cleavages of beta-ethers in radicals might be applied in synthetic reactions. PMID- 16268601 TI - Fluorescence ratiometry of monomer/excimer emissions in a space-through PET system. AB - [Reaction: see text]. Fluorogenic calix[4]arenes (1 and 2) bearing a pendent ethyleneamine on their triazacrown rings, respectively, were synthesized in the cone conformation. Compared with 4, free 1 and 2 display a relatively weak emission, reflecting that a PET process from the pendent amine group (-CH2CH2NH2) to the fluorogenic pyrenes is mainly operated. Addition of various metal ions or anions to the solution of 1 or 2 reduces the PET because the pendent alkylamine takes part in the complexation, causing their fluorescence spectra to be changed. When Pb2+, a quenching metal ion, is added to 1 or 2, their pyrene monomer emission is enhanced with their excimer emission quenched, which is due to conformational changes of the facing carbonyl groups as well as to the participation of the ethyleneamine into the three-dimensional Pb2+ ion encapsulation. In contrast, upon addition of alkali metal ions to the 1 and 2, both monomer and excimer emissions are observed to increase, which is attributable to the CHEF effect and the retained conformations. For anion sensing, both 1 and 2 show a high selectivity for F- ions over other anions tested. When the F- ion is bound to 1 or 2 by hydrogen bonding between the amide NH of the triazacrown ring and F-, both their monomer and excimer emissions are weakened due to PET from the bound F- to the pyrene units. PMID- 16268603 TI - Photoinduced hinge-like molecular motion: studies on xanthene-based cyclic azobenzene dimers. AB - [Structure: see text]. Molecular devices incorporating azobenzene units represent active components of smart systems, as they are capable of exhibiting photoregulated cooperative molecular motion. Herein, we describe the synthesis, X ray crystal analysis, and photochemical and thermal studies of a xanthene based cyclic azobenzene dimer and its precursor. The trans-trans isomer of the azobenzene dimer upon photoirradiation transforms to the cis-cis isomer through an intermediate trans-cis isomer. The X-ray crystal structures of the trans-trans isomer (open) and the cis-cis isomer (closed) provide unambiguous proof for the hinge-like molecular motion in this class of molecules. The inferences drawn from photochemical and thermal studies shed light on the effect of varied substitution and cyclic structures on the different transitions. The lifetime of the cis-cis isomer is estimated to be 6.43 years, whereas the trans-cis isomer is short-lived (2.73 min) at 303 K. A rational explanation for the relative stability of the different isomers is derived from the isokinetic plot and theoretical calculations. PMID- 16268604 TI - From cyclopentanone oximes to bis[1,2,3]dithiazolo-s-indacenes, cyclopenta[c][1,2]thiazine, pentathiepino-, tetrathiino-, and thienocyclopenta[1,2,3]dithiazoles as a rich source of new materials. AB - [Structure: see text]. The 1,5- and 1,7-s-hydrindacenedione dioximes reacted with S2Cl2 and iBu3N to give the first examples of bis[1,2,3]dithiazolo-s-indacenes; one of them was a near-infrared dye. In contrast, the silylated bicyclo[3.3.0]octan-2,6-dione dioxime reacted with S2Cl2 and Et3N to give a bicyclic 4-cyanoethylcyclopenta[1,2,3]dithiazole or, after addition of Li2S, a tricyclic 4-cyanoethyl-5,6-pentathiepinocyclopenta[1,2,3]dithiazole, also obtained from 2-cyanoethylcyclopentanone oxime, S2Cl2, and Hunig's base. In related reactions, 2-oxocyclopentylpropionate oxime gave the expected cyclopenta[1,2,3]dithiazole, in addition to an unexpected cyclopenta[c][1,2]thiazine that showed a reversible reduction wave in its CV at 0.95 V. Ethyl 2-oxocyclopentanecarboxylate oxime reacted with S2Cl2, Hunig's base, and Li2S to give a 5,6-tetrathiinocyclopenta[1,2,3]dithiazole derivative. Cyclopentathiophen-4-one oximes reacted with S2Cl2 and iBu3N to give thienocyclopenta[1,2,3]dithiazoles that showed UV-vis spectral bands that depended on the positions of the ring fusion. PMID- 16268605 TI - Koopmans-based analysis of the optical spectra of p-phenylene-bridged intervalence radical ions. AB - [Figure: see text]. The optical spectra of 10 p-phenylene-bridged delocalized intervalence compounds MC6H4M*- or *+ are analyzed using the Koopmans-based method, which considers only transitions from filled orbitals to the singly occupied orbital (SOMO), called Hoijtink type A transitions, and from the SOMO to unoccupied orbitals, Hoijtink type B transitions, and ignores configuration interaction. The radical ions with quinonoid structures, those that form ring-M double bonds with M = C(CN)2, NMe2, 3-oxo-9-azabicyclo[3.3.1], NPPh3, and O when the odd electron of the intervalence oxidation level is removed, are calculated to have the lowest-allowed type B transition lying mostly above the lowest allowed A transition, with B(i)- A(j) decreasing in the order shown from +14 370 to -1390 cm(-1), and the more intense second-lowest-allowed type B transition B(i) - A(j) from +14 940 to +7070 cm(-1). The five radical anions with benzenoid structures, which form ring-M single bonds with X = CN, CO2Me, CHO, C3HMeBF2O2, and NO2 when the odd electron of the intervalence oxidation level is removed, have a B(i)- A(j) value of the opposite sign that increases in magnitude from 2880 to -17 050 cm(-)(1) in the order shown. Configuration interaction is of course present in the observed spectra, and the predictions ignoring it mostly overestimate transition energies by 1900-2600 cm(-1) for the quinonoid compounds (but by 450 cm(-1) for the M = C(CN)2 radical anion), and by 1000-1400 cm(-1) for the benzenoid compounds (2500 cm(-1) for the M = CN radical anion). The very simple Koopmans-based model is useful for considering the optical spectra of these radical ions. PMID- 16268606 TI - Dynamic chirality in donor-acceptor pretzelanes. AB - [Figure: see text]. A series of donor-acceptor pretzelanes has been synthesized, using self-assembly and template-directed protocols, and the dynamic processes that these pretzelanes undergo have been investigated in solution. These compounds exist as libraries of diastereoisomers as a result of their multiple stereoelements, which are dynamically interconverted by several different, in some cases competing, processes. Altering the structure of the pretzelanes changes the rates and mechanisms by which these diastereoisomers equilibrate. Additionally, inserting an element of fixed chirality allows the equilibrium to be biased, while maintaining the barrier to the equilibration processes. These results bode well for the future construction of molecular devices based on switchable diastereoisomerism involving metastable states. PMID- 16268607 TI - Theoretical analysis of the high versatility in PtCl2-mediated cycloisomerization of enynes on a common mechanistic basis. AB - [Reaction: see text]. Transformations of enynes upon treatment with electrophilic transition-metal complexes, such as PtCl2, are strongly substrate-dependent processes and may yield a wide variety of cyclic compounds. Despite the high versatility, many of these processes could be closely related from a mechanistic point of view. Theoretical analyses of the plausible reaction mechanisms provide support for a unified mechanistic picture based on the electrophilic activation of the triple bond by the catalyst, which triggers the nucleophilic alkene attack through an endo- or exo-cyclization mode, to form the cyclopropylcarbene species. Then, these common key intermediates may evolve through alternative paths to afford a range of cyclic compounds. The preference for each path and the evolution of these intermediates are governed by the nature of the starting enyne. The effects induced by different structural motifs, such as the role played by a heteroatom directly attached to the triple bond, the tether length, the substitution on the acetylenic position, and the gem-dialkyl substitution on the tether, among others, are discussed. The proposed common mechanistic scheme can rationalize and account for the experimental observations accumulated. PMID- 16268608 TI - Regio- and stereoselective reactions of gem-difluorinated vinyloxiranes with heteronucleophiles. AB - [Reaction: see text]. Regio- and stereoselectivity in reactions of gem difluorinated vinyloxiranes with heteronucleophiles were successfully controlled. Halogen atoms were introduced regioselectively at the allylic epoxide carbon with an inversion in stereochemistry using MgBr2*Et2O or Li2CuCl4 to produce anti-vic halohydrine. The other diastereomers were obtained selectively using LiBr/AcOH or BCl3, and SN2' type products were formed selectively with excellent E preference by changing the reaction temperature. Moreover, a further investigation led us to find that a regio- and stereoselective SN2' addition of several Brnsted acids was dependent on the pKa values of the acids. Under strong acidic conditions, we exclusively obtained E allylic alcohols. PMID- 16268609 TI - Kumada coupling of aryl and vinyl tosylates under mild conditions. AB - [Reaction: see text]. Aryl and alkenyl tosylates are easily prepared, inexpensive and, thus, attractive for transition-metal-catalyzed couplings, but their reactivity is low. We report examples of mild, palladium-catalyzed coupling of aryl, alkenyl, and alkyl Grignard reagents with aryl and alkenyl tosylates. The resulting biaryls, vinylarenes, and alkylarenes were isolated in good to excellent yield. These couplings were conducted with a nearly equimolar ratio of the two reactants, and many examples were conducted at room temperature. PMID- 16268610 TI - Synthetic utilization of polynitroaromatic compounds. 3. Preparation of substituted dibenz[b,f][1,4]oxazepine-11(10H)-ones from 2,4,6-trinitrobenzoic acid via nucleophilic displacement of nitro groups. AB - [Reaction: see text]. 1,3-dinitrodibenz[b,f][1,4]oxazepin-11(10H)-one, prepared by intramolecular displacement of nitro group in N-(2-hydroxyphenyl)-2,4,6 trinitrobenzamide, reacts with O- and S-nucleophiles to yield the products of mono- or bis-substitution of the nitro groups. The nitro group in position 3 is displaced first. This observation is in contrast with earlier results for the nitro-substituted benzoannulated five-membered heterocycles. This difference in reactivity is likely due to the increased steric hindrance for peri-nitro group displacement in the case of the benzoannulated seven-membered heterocycle. N Alkylation of the nitro-substituted dibenz[b,f][1,4]oxazepin-11(10H)-ones yields analogues of a known antidepressant drug Sintamil. The structure of the products is confirmed by NOE experiments and alternative synthesis. PMID- 16268611 TI - Polyfunctional tetrahydropyrido[2,3-b]pyrazine scaffolds from 4-phenylsulfonyl tetrafluoropyridine. AB - [Reaction: see text]. Polyfunctional tetrahydropyrido[2,3-b]pyrazine scaffolds can be synthesized by sequential reaction of pentafluoropyridine with sodium phenylsulfinate and an appropriate diamine. The polyfunctionality possessed by the difluorinated tetrahydropyrido[2,3-b]pyrazine scaffolds was demonstrated in selected model reactions with nucleophiles to give access to various polysubstituted [6,6]-ring fused systems. PMID- 16268613 TI - Chirality transfer from guanidinium ylides to 3-alkenyl (or 3-alkynyl) aziridine 2-carboxylates and application to the syntheses of (2R,3S)-3-hydroxyleucinate and D-erythro-sphingosine. AB - [Reaction: see text]. Reaction of chiral guanidinium ylides with alpha,beta unsaturated aldehydes gives 3-(alpha,beta-unsaturated) aziridine-2-carboxylates in high diastereo- and enantioselectivities (up to 93% diastereomeric excess and 98% enantiomeric excess). 3-(1-methylvinyl)- and 3-[(E)-pentadec-1-enyl]aziridine 2-carboxylates were successfully employed to prepare (2R,3S)-3-hydroxyleucinate and d-erythro-sphingosine, respectively. PMID- 16268612 TI - Halichondrin B: synthesis of the C1-C22 subunit. AB - [Reaction: see text]. Two efficient routes to the C1-C22 subunit of halichondrin B are described. The cage ketal 7, which contains 11 asymmetric centers embedded within the ABCDEF-ring framework, was assembled from (+)-conduritol E (27) in 18 steps and 4% overall yield. In a separate route, 7 was also synthesized in 18 steps and 2% overall yield from a derivative of alpha-d-glucoheptonic acid gamma lactone (62). While the former route installs the fully elaborated C-ring endowed with the correct C12 stereochemistry early in the synthesis, the latter features a late-stage introduction of the C12 stereocenter during the ultimate one-pot Michael addition/ketalization cascade to form the CDE-ring system of the cage. The importance of the C12 stereocenter to the crucial ketalization event is discussed through comparison of these two strategies. PMID- 16268614 TI - Electroreductive acylation of aromatic ketones with acylimidazoles. AB - [Reaction: see text]. The intermolecular reductive coupling of aromatic ketones with acylimidazoles was effected by electroreduction in the presence of chlorotrimethylsilane and gave alpha-trimethylsiloxy ketones and esters. The best result was obtained using Bu4NPF6 as a supporting electrolyte and a Pb cathode in THF. The alpha-trimethylsiloxy-containing products were transformed to the corresponding alpha-hydroxy ketones and esters by treatment with TBAF in THF. This method was also effective for the intramolecular reductive coupling of delta and epsilon-keto acylimidazoles. PMID- 16268615 TI - Selective ring-opening of nonactivated amino aziridines by thiols and unusual nucleophilic substitution of a dibenzylamino group. AB - [Reaction: see text]. The reaction of chiral 2-(1-aminoalkyl)aziridines 1 with different thiols, in the presence of BF3*Et2O, is reported. The obtained products were dependent on the structure of the starting amino aziridines 1. Thus, enantiopure (2S,3S)-2-(alkylthio)alkane-1,3-diamines 2 were obtained from aziridines with C-2 substituents with lower steric congestion and partially racemized (2S,3S)-2,3-bis(alkylthio)alkan-1-amines 3 (ee = 56-66%) from aziridines with larger C-2 subtituents. In both cases, the opening of the nonactivated aziridine ring at C-2 took place with retention of configuration and proceeded with regio- and stereoselectivity at C-2. In the synthesis of 3, 2 equiv of thiol reacts with 1 and the opening of aziridine ring at C-2 was followed by an unusual displacement of the dibenzylamino group by a second equivalent of thiol. The regiochemistry and relative configuration of compounds 3 was established by single-crystal X-ray analysis. A mechanism is proposed to explain the results obtained. PMID- 16268616 TI - Three-carbon Dowd-Beckwith ring expansion reaction versus intramolecular 1,5 hydrogen transfer reaction: a theoretical study. AB - [Reaction: see text]. The evolution of the primary radicals formed by addition of AIBN/HSnBu3 to methyl 1-(3-iodopropyl)-5-oxocyclopentanecarboxylate, methyl (1R,2R)-1-(3-iodopropyl)-2-methyl-5-oxocyclopentanecarboxylate, and methyl (1R,2S)-1-(3-iodopropyl)-2-methyl-5-oxocyclopentanecarboxylate in benzene has been theoretically investigated by ROMP2/6-311++G(2d,2p)//UB3LYP/6-31G(d,p) calculations taking into account the effect of solvent through a PCM-UAHF model. According to the theoretical results, for methyl 1-(3-iodopropyl)-5 oxocyclopentanecarboxylate and methyl (1R,2S)-1-(3-iodopropyl)-2-methyl-5 oxocyclopentanecarboxylate the major product is the cyclooctane derivative from the three-carbon ring expansion, whereas for methyl (1R,2R)-1-(3-iodopropyl)-2 methyl-5-oxocyclopentanecarboxylate the major product is that corresponding to the 1,5-H transposition in agreement with the experimental findings. This different behavior is a consequence of several factors determining the relative energy barriers. The methyl substituent destabilizes the ring expansion process for methyl (1R,2R)-1-(3-iodopropyl)-2-methyl-5-oxocyclopentanecarboxylate because of steric repulsion but favors it in the case of the beta-trans-substituted substrate because it makes possible the evolution of the system along more favorable conformations. The methyl group also favors the 1,5-H transposition rendering the transposed product a tertiary radical. The second stage of the ring expansion process is stabilized by resonance. PMID- 16268617 TI - Asymmetric transfer hydrogenation of ketones catalyzed by hydrophobic metal-amido complexes in aqueous micelles and vesicles. AB - [Reaction: see text]. Asymmetric transfer hydrogenation of ketones, especially alpha-bromomethyl aromatic ketones, catalyzed by unmodified, hydrophobic transition metal-amido complexes (TsDPEN-M), was performed successfully with significant enhancement of activity, chemoselectivity, and enantioselectivity (up to 99% ee) in aqueous media containing micelles and vesicles. The hydrophobic catalyst, embedded in micelles constructed from the surfactant cetyltrimethylammonium bromide (CTAB), could be separated from the organic phase along with the products and was recycled for at least six times. PMID- 16268618 TI - Absolute stereochemistry of citrinadins a and B from marine-derived fungus. AB - [Structure: see text]. Citrinadin A (2) is a pentacyclic indolinone alkaloid isolated from the cultured broth of a fungus, Penicillium citrinum, which was separated from a marine red alga. The absolute stereochemistry of the pentacyclic core in 2 and its new congener, citrinadin B (1), was elucidated by analysis of the ROESY spectrum for the chlorohydrin derivative (3) of 1 as well as comparison of the electronic circular dichroism (ECD) spectra for 1 and 2 with those of known spirooxiindole alkaloids. On the other hand, the absolute configuration at C-21 bearing an epoxide ring was assigned as S by comparison of the vibrational circular dichroism (VCD) spectra of 1 with those of model compounds 2S- and 2R 2,3-epoxy-3,3-dimethyl-1-phenylpropan-1-one (4a and 4b, respectively). PMID- 16268619 TI - Efficient synthesis and characterization of novel bibenzimidazole oligomers and polymers as potential conjugated chelating ligands. AB - [Structure: see text]. A simple and mild condensation route for the synthesis of novel bibenzimidazole oligomers and polymers is reported here using methyl 2,2,2 trichloroacetimidate as a key starting material. The dimer, trimer, tetramer, and polymers of bibenzimidazole were synthesized as a new series of potential conjugated chelating ligands for metallopolymer studies. The polymers show a maximum absorption at around 400 nm. The optical band gap of the polymer was estimated to be 2.68 eV. PMID- 16268620 TI - Synthesis of all 16 stereoisomers of pinesaw fly sex pheromones--tools and tactics for solving problems in fluorous mixture synthesis. AB - [Reaction: see text]. The application of fluorous mixture synthesis (FMS) for accessing natural products and their stereoisomers was validated by the total synthesis of all 16 stereoisomers of the pine sawfly sex pheromone. Four fluorous p-methoxybenzyl groups were used as tags, and a "4-mix-4-split" approach was employed in a divergent synthesis. This paper presents the details of the FMS of pine sawfly sex pheromones with an emphasis on identification and solving of problems encountered when working with fluorous mixtures. PMID- 16268621 TI - Efficient regioselective synthesis of 6-amino-5-benzoyl-1-substituted 2(1H) pyridinones. AB - [Reaction: see text]. A regioselective and efficient approach toward 6-amino-5 benzoyl-1-substituted 2(1H)-pyridinones by reaction of acyclic ketene aminals with propiolic acid ester was developed. The effect of the solvent and temperature on the regioselectivity of the reaction and the compatibility of the target compounds to functional group manipulations was examined. Substrates with an ortho substituent build atropisomers due to the restricted rotation around the C-N bond. The enantiomers were separated, and the barrier of rotation was determined experimentally. Quantum chemical calculations allowed a ranking of the barrier heights, and a new mechanism of rotation by deformation of the central pyridinone moiety is proposed. PMID- 16268622 TI - Asymmetric phase-transfer catalyzed glycolate alkylation, investigation of the scope, and application to the synthesis of (-)-ragaglitazar. AB - [Reaction: see text]. Asymmetric glycolate alkylation using a protected acetophenone surrogate under solid-liquid phase-transfer conditions is a new approach to the synthesis of 2-hydroxy esters and acids. Diphenylmethyloxy-2,5 dimethoxyacetophenone 1 with a trifluorobenzyl cinchonidinium bromide catalyst 9 (10 mol %) and cesium hydroxide provided S-alkylation products 2 at -35 degrees C in high yield (80-99%) and with excellent enantioselectivities using a wide range of electrophiles (80-90% ee). Alkylated products were elaborated to useful alpha hydroxy intermediates 3 using bis-TMS peroxide Baeyer-Villiger conditions and selective transesterification reactions. The ester products have been enantioenriched by simple recrystallization from ether to give a single isomer (99% ee). A tight ion-pair model is proposed for the observed S-stereoinduction that includes van der Waals contacts between the extended enolate and the isoquinoline of the catalyst. To demonstrate the utility of the new methodology, the anti-diabetes drug (-)-ragaglitazar 24 was synthesized in six steps from a key 2-alkoxy-3-p-phenoxypropionic acid 26 that was made using PTC glycolate alkylation. PMID- 16268623 TI - Synthesis of three marine natural sesterterpenolides from methyl isoanticopalate. First enantioselective synthesis of luffolide. AB - [Reaction: see text]. The synthesis of three marine sponge metabolites, luffolide (4), 5, and 6, are described for the first time, establishing the absolute configuration of these compounds. The key intermediate, aldehyde 17, was obtained from methyl isoanticopalate, 11. The addition of 3-furyllithium to 17 and subsequent photochemical oxidation give the gamma-hydroxybutenolide 5 and its epimer at C-16. Sesterterpenolide 6 is obtained by dehydration of 5. From the key aldehyde 17, luffolide (4) was obtained in six steps. PMID- 16268624 TI - Asymmetric synthesis of tetrahydropalmatine via tandem 1,2-addition/cyclization. AB - [Reaction: see text]. The enantioselective synthesis of both enantiomers of tetrahydropalmatine (2) (ee = 98%), a natural alkaloid belonging to the tetrahydroprotoberberine family, is described. The key step of this total synthesis is based on our tandem 1,2-addition/ring-closure methodology employing lithiated methylbenzamide and benzaldehyde SAMP or RAMP hydrazones as substrates. An initial route was investigated for the formation of N- and 3-substituted dihydroisoquinolones starting from 2-substituted benzaldehyde SAMP hydrazones, but although high diastereoselectivity was achieved, only disappointing yields were obtained. In our subsequent synthetic strategy, 2,3-dimethoxy-6 methylbenzamide 6 and 3,4-dimethoxybenzaldehyde SAMP or RAMP hydrazone 19 gave the dihydroisoquinolones 20 in high diastereomeric purity (de > or = 96%) and reasonable yield (54-55%), taking into account the complex functionalities established in one step. Cleavage of the N-N bond of the chiral auxiliary and reduction of the carbonyl group of the amide moiety were performed in the same step, and the resulting tetrahydroisoquinolines 22 (ee = 99%) were N functionalized by treatment with various electrophiles to investigate the ring closure by Pummerer, Friedel-Crafts, and Pomeranz-Fritsch reactions. The Pummerer cyclization led to the formation of (S)-(-)-2 with slight racemization (ee = 89%), whereas the Friedel-Crafts reaction proved to be unsuccessful. Finally, Pomeranz-Fritsch-type cyclization afforded the desired title compound (R)-(+)-2 in excellent enantioselectivity in 9% overall yield over seven steps and after optimization of the last step (S)-(-)-2 in 17% overall yield. PMID- 16268625 TI - Z/E-photoisomerizations of olefins with 4npi- or (4n + 2)pi-electron substituents: zigzag variations in olefin properties along the T(1) state energy surfaces. AB - [Figure: see text]. A quantum chemical study has been performed to assess changes in aromaticity along the T1 state Z/E-isomerization pathways of annulenyl substituted olefins. It is argued that the point on the T1 energy surface with highest substituent aromaticity corresponds to the minimum. According to Baird (J. Am. Chem. Soc. 1972, 94, 4941), aromaticity and antiaromaticity are interchanged when going from S0 to T1. Thus, olefins with S0 aromatic substituents (set A olefins) will be partially antiaromatic in T1 and vice versa for olefins with S0 antiaromatic substituents (set B olefins). Twist of the C=C bond to a structure with a perpendicular orientation of the 2p(C) orbitals (3p*) in T1 should lead to regaining substituent aromaticity in set A and loss of aromaticity in set B olefins. This hypothesis is verified through quantum chemical calculations of T1 energies, geometries (bond lengths and harmonic oscillator measure of aromaticity), spin densities, and nucleus independent chemical shifts whose differences along the T1 PES display zigzag dependencies on the number of -electrons in the annulenyl substituent of the olefin. Aromaticity changes are reflected in the profiles of the T1 potential energy surfaces (T1 PESs) for Z/E-isomerizations because olefins in set A have minima at 3p* whereas those in set B have maxima at such structures. The proper combination (fusion) of the substituents of set A and B olefins could allow for design of novel optical switch compounds that isomerize adiabatically with high isomerization quantum yields. PMID- 16268626 TI - Self-inclusion behavior and circular dichroism of aliphatic chain-linked beta cyclodextrin-viologen compounds and their reduced forms depending on the side of modification. AB - [Reaction: see text]. The self-inclusion behavior and induced circular dichroism (ICD) characteristics of two beta-cyclodextrin (beta-CD) derivatives, in which a 1-methyl-4,4'-bipyridinium (viologen) group is connected by an octamethylene chain to either the primary (2(2+)) or secondary (3(2+)) side of beta-CD, and of their reduced forms, are investigated. 1H NMR studies showed that 2(2+) forms an intramolecular self-inclusion complex with K(in) = 3.1 +/- 0.4, whereas 3(2+) forms a head-to-head type of dimer with K(D) = 65 +/- 10 M(-1) at 25 degrees C. 2(2+) and 3(2+) form [2]pseudorotaxanes with alpha-CD, with the secondary side of the alpha-CD facing the viologen moiety. The ICD characteristics of mono-6-[4-(1 methyl-4-pyridinio)-1-pyridinio]-beta-CD (1(2+)), 2(2+), 3(2+), and methyloctyl viologen-beta-CD complexes were obtained for the oxidized and reduced states of the viologen units. The results indicated dimer formation for 1 degrees , and intramolecular complexation for 2*+ and 2 degrees in which the reduced viologen units are outside the beta-CD cavity. The results also indicated intramolecular complexation for 3*+ and 3 degrees, but with reduced viologen units inside the cavity. This work provides unequivocal evidence of the preference of the secondary side of cyclodextrins for viologen groups, regardless of their oxidation states, and the dependence of ICD of the viologen chromophores on their location with respect to the CD cavity. PMID- 16268628 TI - Kinetic study of the hydrogen abstraction reaction of the benzotriazole-N-oxyl radical (BTNO) with H-donor substrates. AB - [Reaction: see text]. The aminoxyl radical (>N-O*) BTNO (benzotriazole-N-oxyl) has been generated by the oxidation of 1-hydroxybenzotriazole (HBT; >N-OH) with a Ce(IV) salt in MeCN. BTNO presents a broad absorption band with lambda(max) 474 nm and epsilon 1840 M(-1) cm(-1), and spontaneously decays with a first-order rate constant of 6.3 x 10(-3) s(-1) in MeCN at 25 degrees C. Characterization of BTNO radical by EPR, laser flash photolysis, and cyclic voltammetry is provided. The spontaneous decay of BTNO is strongly accelerated in the presence of H-donor substrates such as alkylarenes, benzyl and allyl alcohols, and alkanols, and rate constants of H-abstraction by BTNO from a number of substrates have been spectroscopically investigated at 25 degrees C. The kinetic isotope effect confirms the H-abstraction step as rate-determining. Activation parameters have been measured in the 15-40 degrees C range with selected substrates. A correlation between E(a) and BDE(C-H) (C-H bond dissociation energy) for a small series of H-donors has been obtained according to the Evans-Polanyi equation, giving alpha = 0.44. From this plot, the experimentally unavailable BDE(C-H) of benzyl alcohol can be extrapolated, as ca. 79 kcal/mol. With respect to the H abstraction step, peculiar differences in the DeltaS++ parameter emerge between an alkylarene, ArC(H)R2, and a benzyl alcohol, ArC(H)(OH)R. The data acquired on the H-abstraction reactivity of BTNO are compared with those recently reported for the aminoxyl radical PINO (phthalimide-N-oxyl), generated from N hydroxyphthalimide (HPI). The higher reactivity of radical PINO is explained on the basis of the higher energy of the NO-H bond of HPI, as compared with that of HBT (88 vs ca. 85 kcal/mol, respectively), which is formed on H-abstraction from the RH substrate. PMID- 16268627 TI - Total synthesis of Bengamide E and analogues by modification at C-2 and at terminal olefinic positions. AB - [Reaction: see text]. The total synthesis of the natural product Bengamide E, one of the members of a new class of antitumor natural products of marine origin, is reported based on a convergent and flexible synthetic route featuring an oxirane ring-opening reaction and an olefin cross metathesis. In a similar way, analogues structurally modified at C-2 and at the terminal vinyl positions were prepared by introduction of various nucleophiles and alkyl substituents during the epoxide opening and the olefin cross metathesis steps, respectively. These studies demonstrate the validity of our synthetic strategy, although they reveal some problems associated with the olefin cross metathesis, whose efficiency depends on the substituent at the C-2 position as well as the steric environment of the alkene. PMID- 16268629 TI - Investigation of the Diels-Alder cycloadditions of 2(H)-1,4-oxazin-2-ones. AB - [Reaction: see text]. A variety of 5-chloro-2(H)-1,4-oxazin-2-ones bearing a range of substituents at their 3- and 6-positions undergo Diels-Alder cycloaddition as a 2-azadiene component with electron-rich, electron-deficient, and electron-neutral dienophiles. These reactions proceed with moderate regio- and stereoselectivity to afford relatively stable and readily isolable bridged bicyclic lactone cycloadducts. Chemical manipulation of these cycloadducts affords highly substituted and functionally rich piperidines. The regio- and stereochemical preferences of the cycloadditions of 5-chloro-2(H)-1,4-oxazin-2 ones are investigated computationally using density functional theory (B3LYP/6 31G). PMID- 16268630 TI - Rh-catalyzed enantioselective diboration of simple alkenes: reaction development and substrate scope. AB - [Reaction: see text]. The rhodium-catalyzed reaction between bis(catecholato)diboron and simple alkenes results in the syn addition of the diboron across the alkene. The resulting 1,2-bis(boronate) is subsequently oxidized to provide the 1,2-diol. In the presence of enantiomerically enriched Quinap ligand, high enantioselection in the diboration can be achieved. The reaction is highly selective for trans- and trisubstituted alkenes and can be selective for some monosubstituted alkenes as well. The development of this reaction is described as is the substrate scope and experiments that are informative about the reaction mechanism and competing pathways. PMID- 16268631 TI - A highly regio- and stereoselective nickel-catalyzed ring-opening reaction of alkyl- and allylzirconium reagents to 7-oxabenzonorbornadienes. AB - [Reaction: see text]. An efficient method for the synthesis of cis-2-alkyl- or allyl-1,2-dihydronaphthalenes via a nickel-catalyzed highly regio- and stereoselective ring-opening addition of alkyl- or allylzirconium reagents to 7 oxabenzonorbornadienes is described. Treatment of 7-oxabenzonorbornadienes 1a-c with various alkylzirconium reagents 2a-j (Cp2ZrClCH2CH2R: R = tert-butyl, n butyl, n-pentyl, -(CH2)3CH=C(CH3)2, -SiMe3, -CH2SiMe3, -(CH2)3Br, cyclopentyl, cyclohexyl, and benzyl) in the presence of NiBr2(dppe) and Zn powder in dry THF at 50 degrees C afforded the corresponding cis-2-alkyl-1,2-dihydronaphthalene derivatives 3a-m in good yields. In addition, allyl zirconium reagents 4a-c also underwent ring-opening reactions with 1a and 1c to give 5a-d in very good yields. The alkylative ring-opening products from 7-oxabenzonorbornadiene can be further converted to naphthalene derivatives 6a-c, via an acid-mediated dehydration, in good to excellent yields. A possible mechanism for the present catalytic reaction was proposed. PMID- 16268632 TI - Chemoenzymatic dynamic kinetic resolution of acyloins. AB - [Reaction: see text]. Acyloins (alpha-hydroxy ketones) are important building blocks in organic synthesis, e.g., for the total synthesis of epothilones. Optically pure acyloins can be obtained by lipase-catalyzed kinetic resolution (KR) of the racemate with, for example, Burkholderia cepacia lipase, but this process suffers from a yield limitation of 50%. To devise a dynamic kinetic resolution (DKR), we studied the racemization of two different acyloins and corresponding esters with various amine bases and ion exchangers. No combination of base and solvent was found that could selectively racemize the acyloin or corresponding ester under the conditions needed for a DKR. In contrast to bases, acidic resins (ARs) were found to racemize the acyloins selectively in n-hexane and in water. Unfortunately, the AR deactivated the lipase, preventing a one-pot DKR. Minor side reactions involving the AR, the substrate acyloin, and the vinyl ester acyl donor were also observed. However, an efficient DKR was made possible by the spatial separation of lipase and ion exchanger, with enzymatic transesterification and AR-catalyzed racemization taking place simultaneously in two compartments connected by a pump loop. The conversion of substrate alcohol was 91%, the selectivity toward the product butyrate ester 90%, and the enantiomeric excess of the (S)-product 93% ee. PMID- 16268633 TI - Ferrocenylketene and ferrocenyl-1,2-bisketenes: direct observation and reactivity measurements. AB - [Structure: see text]. Ferrocenylketene (1) is calculated to be destabilized by 1.6 kcal/mol relative to phenylketene (10) by B3LYP isodesmic comparison to the corresponding alkenes. Ketene 1 generated by Wolff rearrangement in CH3CN is identified by the IR band at 2119 cm(-1) and has a rate constant for reaction with n-BuNH2 less than that for 10 by a factor of 5. 1,2-Bisferrocenyl-1,2 bisketene 18 and 1-ferrocenyl-2-trimethylsilyl-1,2-bisketene 21 were prepared by photochemical ring opening of the corresponding cyclobutenediones, and 18 undergoes rapid ring closure 67 times faster than the corresponding 1,2-diphenyl 1,2-bisketene, while bisketene 21 is longer lived than 18 by a factor of 3.2 x 10(4). PMID- 16268635 TI - Peroxy acid epoxidation of acyclic allylic alcohols. Competition between s-trans and s-cis peroxy acid conformers. AB - [Reaction: see text]. RB3LYP calculations, reported here, indicate that peroxy acid s-cis conformer is more stable than its s-trans counterpart, in agreement with experimental data. Difference in stability is the highest in the gas phase, but it falls considerably on going from the gas phase to moderately polar solvent. In the case of peroxy formic acid, the enthalpy (free energy) difference is about 3.4 (2.5) kcal/mol, respectively, in the gas phase but decreases to 1.2 (0.6) kcal/mol in dichloromethane solution. Introduction of an alkyl or aryl substituent on the peroxy acid, that is, on passing to peroxy acetic, peroxy benzoic (PBA), and m-chloroperoxy benzoic acid (MCPBA), adds a further significant (1.0-1.5 kcal/mol) favor to the s-cis isomer. RB3LYP/6-31+G(2d,p) calculations on the epoxidation of 2-propenol with peroxy formic and peroxy benzoic acids, respectively, suggest that the less stable peroxy acid s-trans conformer can compete with the more stable s-cis form in epoxidation reaction of these substrates. Transition structures arising from s-trans peroxy acids ("trans" TSs) retain both the well-established, for "cis" TS, perpendicular orientation of the O-H peroxy acid bond relative to the C=C bond and the one-step oxirane ring formation. These TSs collapse to the final epoxide via a 1,2-H shift at variance with the 1,4-H transfer of the classical Bartlett's "cis" mechanism. The "trans" reaction pathways have a higher barrier in the gas phase than the "cis" reaction channels, but in moderately polar solvents they become competitive. In fact, the "trans" TSs are always significantly more stabilized than their "cis" counterparts by solvation effects. Calculations also suggest that going from peroxy formic to peroxy benzoic acid should slightly disfavor the "trans" route relative to the "cis" one, reflecting, in an attenuated way, the decrease in the peroxy acid s-trans/s-cis conformer ratio. The predicted behavior for MCPBA parallels that of PBA acid. PMID- 16268634 TI - Synthesis and luminescence of soluble meso-unsubstituted tetrabenzo- and tetranaphtho[2,3]porphyrins. AB - [Structure: see text]. Syntheses of soluble tetrabenzoporphyrins (TBP) and tetranaphtho[2,3]porphyrins (TNP), with multiple substituents in the conjugated aromatic rings but bearing no substituents in the meso-positions, is reported. Both types of porphyrins were obtained by direct aromatization of precursor porphyrins, annealed with either cyclohexene or dihydronaphthalene fragments. TBPs and TNPs possess powerful absorption bands in the near-infrared (lambda = 610-710 nm, epsilon = 100,000-300,000 M(-1) cm(-1)) and exhibit strong luminescence. Free bases and Zn complexes fluoresce with quantum yields of up to 50%, whereas Pd and Pt complexes phosphoresce in solutions at ambient temperatures. Remarkably, the phosphorescence quantum yields of Pd and Pt TBPs reach as high as 20-50%, which places them among the brightest near-infrared phosphors known to date. PMID- 16268636 TI - Effect of E-ring modifications in camptothecin on topoisomerase I inhibition: a quantum mechanics treatment. AB - Camptothecins (CPTs) are the prototypical class of topoisomerase I (Top1) inhibitors with significant anticancer activities. Structure-activity relationship studies have demonstrated that inverting the stereochemistry at C-20 (R-CPT) or changing the E-ring lactone to a lactam (CPT-lactam) abolishes the Top1 inhibitory activity. The explanations that have been advanced for these effects are that there is either a failure of hydrogen bond formation involving the C-20 hydroxyl group of R-CPT or a failure of E-ring opening of the lactam, which have been proposed to be required for Top1 inhibition. We demonstrate here that the preferred conformation for the CPTs has the 20-Et pseudoaxial, while the 20-OH is pseudoequatorial, and therefore, the 20-OH groups in all the three CPT analogues (S-CPT, R-CPT, and CPT-lactam) are able to hydrogen bond with Asp533. The loss of the Top1 inhibitory activity by the latter two CPT analogues is attributed to the decreased pi-pi stacking interaction energy with the neighboring base pairs compared to the natural S-CPT. The differences in pi-pi stacking interaction energies are derived from the differential electrostatics on the E-ring. PMID- 16268637 TI - Efficient synthesis of [3H]-sanglifehrin A via selective oxidation/reduction of alcohols at C31 and C35. AB - [Reaction: see text]. Sanglifehrin A is a novel complex natural product showing strong immunosuppressive activity and remarkably high affinity for cyclophilin A. To assess its pharmacokinetic properties in vivo, an efficient synthetic route was developed to introduce a tritium label in position C35 of sangliferin A via an oxidation/reduction strategy. The synthetic approach is particularly attractive, because the C35-oxo intermediate 7 is available in good yield on large scale and the reducing agent, lithium tri-sec-butylborotritide, is readily available. An attempt to apply a similar strategy to the alcohol in position C31 led primarily to C31-epi-hydroxy sanglifehrin A under a variety of conditions. PMID- 16268638 TI - Synthesis of 2-nitro- and 2,2'-dinitrobiphenyls by means of the suzuki cross coupling reaction. AB - [Reaction: see text]. Mechanistic investigations and protocols for the synthesis of 2-nitrobiphenyls and 2,2'-dinitrobiphenyls are disclosed. It is revealed that obstacles appear during the transmetalation step when the phenylboronic acid is substituted with a nitro group in the 2-position, whereas when substituted in the 3- or 4-positions, the reaction follows similar patterns as found in the electrophilic substitution of nitrobenzenes, an observation that may be attributed to the elimination step of the catalytic cycle. PMID- 16268639 TI - Ring closure to beta-turn mimics via copper-catalyzed azide/alkyne cycloadditions. AB - [Structure: see text]. Copper-catalyzed azide alkyne cycloadditions of the linear substrates 1 were used to form the cyclic derivatives 2. Computational, NMR, and CD analyses of these compounds indicate that their most favorable conformational states include type I and type II beta-turn conformations. Selectivity for the dimeric products 6 in these cyclization reactions is discussed. PMID- 16268640 TI - Investigating the reaction of N-carbobenzoxy-O-carbobenzoxy- hydroxylamine with dimethyl sulfoxide: formation of S,S-dimethyl-N-[(phenylmethoxy)- carbonyl]sulfoximine. AB - [Reaction: see text]. N-carbobenzoxy-O-carbobenzoxyhydroxylamine (1a) underwent a thermally induced reaction in DMSO in which there is net N-alpha-eliminative oxidation with tandem oxidative incorporation of DMSO to yield S,S-dimethyl-N [(phenylmethoxy)carbonyl]sulfoximine. Mechanisms for the formation of the sulfoximine are presented as well as the product characterizations, including the X-ray crystal structure. PMID- 16268641 TI - Highly effective fluorescent and colorimetric sensors for pyrophosphate over H2PO4- in 100% aqueous solution. AB - [Structure: see text]. This study demonstrated that Zinpyr-1*Zn2+ acts as a fluorescent and colorimetric sensor for pyrophosphate at pH 7.4. In addition, Zinpyr-1*Cu2+ and DIARB-1*Cu2+ complexes were found to act as selective fluorescent sensors for pyrophosphate. Furthermore, the chemosensors Zinpyr 1*Zn2+ and Zinpyr-1*Cu2+ show highly selective and ratiometric fluorescence changes for pyrophosphate compared with H2PO4-. PMID- 16268642 TI - Palladium-catalyzed silylations of hydrosilanes with aryl halides using bulky alkyl phosphine. AB - [Reaction: see text]. A palladium-catalyzed direct trialkylsilyl transfer to aryl halides has been developed. In the presence of Pd(t-Bu3P)2 and K3PO4, electron rich para- or meta-substitute aryl iodides were coupled efficiently with triethylsilane, triphenylsilane, and dimethylphenylsilane to afford the corresponding silylated products in moderate to good yields. PMID- 16268643 TI - A simple method for epoxidation of olefins using sodium chlorite as an oxidant without a catalyst. AB - [Reaction: see text]. Sodium chlorite has been demonstrated to be capable of epoxidizing a variety of olefins at 55-65 degrees C (oil bath). Chlorine dioxide is believed to be the pivotal epoxidizing agent in the reaction on the basis of the mechanistic studies. PMID- 16268644 TI - Electrochemical partial fluorination of organic compounds. 80. Synthesis of cyclic alpha-arylthio-alpha-monofluorophosphonate esters. AB - [Reaction: see text]. Seven-membered cyclic alpha-monofluorophosphonate esters such as 2-allyloxy-3-fluoro-3-phenylthio-4,7-dihydro-[1, 2]-oxaphosphinine-2 oxide were successfully synthesized in moderate total yield as 41% from open chain allyl phosphonates having an alpha-arylthio group as an electroauxiliary using an alternative sequence of anodic fluorination and ring-closing olefin metathesis (RCM). On the other hand, in an attempt to synthesize an eight membered analogue, a different type of seven-membered fluorinated cyclic product was formed predominantly by the RCM reaction between the allyloxy groups. PMID- 16268645 TI - Simple, rapid procedure for the synthesis of chloromethyl methyl ether and other chloro alkyl ethers. AB - [Reaction: see text]. Zinc(II) salts catalyze the reaction between acetals and acid halides to provide haloalkyl ethers in near-quantitative yield. Reactions from millimole to mole scale are typically complete in 1-4 h with 0.01 mol % catalyst. The solutions of haloalkyl ethers thus obtained can be utilized directly in reactions in which the presence of the ester byproduct does not interfere. Excess haloalkyl ether is destroyed on workup, thereby minimizing exposure to this class of carcinogenic compounds. PMID- 16268646 TI - Efficient approaches toward the solid-phase synthesis of new heterocyclic azoniaspiro ring systems: synthesis of tri- and tetrasubstituted 10-oxo- 3,9 diaza-6-azoniaspiro[5.5]undecanes. AB - [Reaction: see text]. An efficient approach for the parallel solid-phase synthesis of novel heterocyclic azoniaspiro ring systems is described. The target compounds, the 1,8,9-trisubstituted 10-oxo-3,9-diaza-6-azoniaspiro[5.5]undecanes, were obtained starting from resin-bound reduced dipeptides. The azoniaspiro cation was formed by intramolecular attack of a tertiary nitrogen on pendent alpha-bromocarbonyl. N-3 acylated and N-3 alkylamino carbonyl derivatives of the 1,8,9-trisubstituted 10-oxo-3,9-diaza-6-azoniaspiro[5.5]undecanes were obtained following in solution treatment of the N-3 azoniaspiro derivatives with different carboxylic acids and isocyanates. PMID- 16268647 TI - Synthesis of conformationally constrained cyclic peptides using an intramolecular sonogashira coupling. AB - [Reaction: see text]. Small peptides having a 3-bromobenzyl group at the C termini and n-alkynoyl group at the N-termini undergo a smooth copper-free intramolecular Sonogashira coupling reaction to afford the corresponding cyclic peptides in moderate yields. Scope and limitations of this macrocyclization is demonstrated with di-, tri-, and tetrapeptides. PMID- 16268648 TI - Preparation of substituted pyrimido[4,5-b]-1,4-benzoxazepines, thiazepines, and diazepines via a Pictet-Spengler cyclization. AB - [Reaction: see text]. A synthesis of the title compounds, which have found use as inhibitors of certain receptor tyrosine kinases, was achieved using a Pictet Spengler cyclization as a key step. PMID- 16268649 TI - Contrastive photoreduction pathways of benzophenones governed by regiospecific deprotonation of imidazoline radical cations and additive effects. AB - [Reaction: see text]. In the photoreaction of benzophenones with 1,3-dimethyl-2 phenylbenzimidazoline (DMPBI), benzhydrols were major products. Addition of H2O accelerated the reaction with no change in the product distribution, while AcOH, PhOH, and metal salts such as LiClO4 and Mg(ClO4)2 were effective additives to produce benzpinacols. In contrast, benzpinacols were exclusively formed regardless of the solvent and the additive in the reactions with 2-(o hydroxyphenyl)-1,3-dimethylbenzimidazoline (o-HPDMBI). These observations are consistent with the hypothesis that DMPBI*+ donates a proton at the C2 position to the benzophenone ketyl radicals while o-HPDMBI*+ donates a phenol proton. PMID- 16268650 TI - Cross-metathesis assisted by microwave irradiation. AB - Microwave irradiation effectively accelerates cross-coupling metathesis reactions between deactivated olefins. Reactions have been carried out with the phosphine free Hoveyda-Grubbs catalyst and the "second generation Grubbs' catalyst." While there have been reports that a "microwave effect" is observed in various transformations, the accelerations we observe are due to the efficient and rapid heating and increased pressure in the microwave apparatus. PMID- 16268651 TI - A concise approach to 2-azabicyclo[3.3.1]nonane derivatives from an acyclic precursor. AB - [Reaction: see text]. Condensation of a readily available 5-amino-2-alkenoate ester with alpha-unsubstituted aliphatic aldehydes leads to substituted 1,2,3,4 tetrahydropyridines. Subsequent manipulation of the ester and enamine functions gives a quick access to 2-azabicyclo[3.3.1]nonane-containing compounds. PMID- 16268653 TI - Unexpected and expeditious entry into trifluoromethyl aziridines from substituted beta-dicarbonyl compounds. AB - [Reaction: see text]. One-pot aziridinations were obtained starting from substituted 2,2,2-trifluoroethyl beta-dicarbonyl compounds with nosyloxycarbamates in the presence of an excess of CaO as base. The unexpected ring closure reaction takes place at room temperature, leading to the N-protected alpha-trifluoromethyl aziridines with good yields. The reaction pathway seems to be influenced by the choice of the base. PMID- 16268652 TI - Highly regioselective synthesis of 1-aryl-3 (or 5)-alkyl/aryl-5 (or 3)-(N cycloamino)pyrazoles. AB - [Reaction: see text]. An efficient highly regioselective protocol for the synthesis of isomeric 1,3-diaryl (or 1-aryl-3-alkyl) and 1,5-diaryl (or 1-aryl-5 alkyl)-5 (or 3)-(N-cycloamino)pyrazoles has been reported by cyclocondensation of common alpha-oxoketene N,S-acetal precursors with arylhydrazines by variation of reaction conditions. PMID- 16268654 TI - Pd(II)-bipyridine catalyzed conjugate addition of arylboronic acid to alpha,beta unsaturated carbonyl compounds. AB - [Reaction: see text]. A Pd(II)-catalyzed conjugate addition of arylboronic acid to alpha,beta-unsaturated ketones, aldehydes, esters, etc. in the presence of 2,2'-bipyridine was developed. A mechanism involving transmetalation, insertion of the carbon-carbon double bond into the C-Pd bond, and protonolysis of the resulting C-Pd bond is proposed. The reaction conditions are mild and the yield is high. The presence of 2,2'-bipyridine is crucial for the reaction to inhibit beta-hydride elimination. PMID- 16268656 TI - Synthesis of eupomatilone-6 and assignment of its absolute configuration. AB - [Reaction: see text]. The Zn-mediated Barbier reaction of the biarylaldehyde 8 with crotyl bromide followed by hydroboration and oxidation provided the gamma butyrolactones 4 and 5. The stereoselective installation of methyl group at C-3 by using LiHMDS and MeI completed the synthesis of racemic eupomatilone-6 (2) and its diastereomer 3. The spectroscopic data of 2 was in full agreement with reported spectra of natural product, thus confirming the revised relative configuration of eupomatilone-6. Similarly, an optically active (3R,4R,5S)-isomer of eupomatilone-6 (23) was prepared in which the aldol reaction with thiazolidinethione as a chiral auxiliary was employed as a key step. On the basis of the spectroscopic data and optical rotation values of 23, the absolute configuration of eupomatilone-6 was proposed. PMID- 16268655 TI - Enantiodivergent, biocatalytic routes to both taxol side chain antipodes. AB - [Reaction: see text]. Two enantiocomplementary bakers' yeast enzymes reduced an alpha-chloro-beta-keto ester to yield precursors for both enantiomers of the N benzoyl phenylisoserine Taxol side chain. After base-mediated ring closure of the chlorohydrin enantiomers, the epoxides were converted directly to the oxazoline form of the target molecules using a Ritter reaction with benzonitrile. These were hydrolyzed to the ethyl ester form of the Taxol side chain enantiomers under acidic conditions. This brief and atom-efficient route to both target enantiomers demonstrates both the synthetic utility of individual yeast reductases and the power of genomic strategies in making these catalysts available. PMID- 16268657 TI - Sulfone coupling and double-elimination strategy for carotenoid synthesis. AB - [Reaction: see text]. A highly efficient synthetic method of carotenoid compounds has been developed on the basis of the sulfone coupling and double-elimination strategy. This method highlighted the sulfone-mediated coupling with the novel C(10) dialdehyde, 2,7-dimethyl-4-octenedial, which was easily prepared and efficiently utilized in the synthesis of the conjugated polyene chains. PMID- 16268662 TI - Overtreatment in epilepsy: how it occurs and how it can be avoided. AB - In pharmacotherapy, overtreatment may be defined as an excessive drug load (that is, excessive drug dosages or unnecessary polypharmacy) leading to a suboptimal risk-to-benefit ratio. The risk of overtreatment in the pharmacological management of epilepsy is substantial and may have serious consequences in terms of a greater incidence and severity of adverse effects. These effects can range from subtle CNS impairment to overt toxic effects, including teratogenicity. Overtreatment also causes increased treatment costs and may even lead to a paradoxical deterioration in seizure control. The prevention and correction of overtreatment requires a thorough understanding of the situations and mechanisms that lead to inappropriate prescribing of antiepileptic drugs. These include initiating treatment in conditions where it is not indicated (for example, long term prophylaxis after head trauma or supratentorial surgery in seizure-free patients), use of excessively fast titration rates, prescription of excessively high initial target dosages, failure to consider conditions associated with reduced dosage requirements (for example, old age or comorbidities associated with impaired drug clearance), and failure to consider the dose-response characteristics of the selected drug. Many patients whose seizures do not respond to the initially prescribed medication can be optimally managed by switching to monotherapy with an alternative agent; premature use of combination therapy represents another common form of overtreatment. Overtreatment may also result from a failure to adjust the dosage to prevent or compensate for adverse pharmacokinetic or pharmacodynamic drug interactions, and from a failure to reduce drug load in patients who have not benefited from high dosages or polypharmacy. While the measurement of drug concentrations can aid in minimising adverse effects, there is also a danger of overtreatment resulting from inappropriate interpretation of drug concentration data. Continuation of drug therapy in seizure-free patients in whom the risk-benefit ratio is in favour of gradual withdrawal may also be regarded as overtreatment. Tailoring therapy to the needs of the individual patient is the key to the successful management of epilepsy. Even though the importance of complete seizure control cannot be overemphasised, no patient should be made to suffer more from the adverse effects of treatment than from the manifestations of the seizure disorder. PMID- 16268663 TI - The role of alpha-4 integrin in the aetiology of multiple sclerosis: current knowledge and therapeutic implications. AB - Multiple sclerosis (MS) has been recognised as a disease since the mid-19th century. The delineation of its CNS pathology, revealing the presence of inflammatory demyelination and relative sparing of axons, was originally interpreted as evidence of infection. Despite many studies, a primary infectious aetiology of MS has not been found. However, the occurrence of acute demyelinating disease following a variety of infections and vaccinations, leading to MS in about a third of cases, provides evidence for the existence of an auto allergic pathogenesis for the disease. Improved understanding of the role of the blood-brain barrier in protecting the CNS, and the mechanisms by which cells gain entry into the brain and spinal cord has advanced the understanding of MS. Evidence of the central role of the adhesion molecule alpha4beta1-integrin (very late activation antigen-4 [VLA-4]) for lymphocytes in endothelial transmigration into the CNS specifically, has provided a major insight into the pathogenesis of human demyelinating disease and its experimental model, experimental autoimmune encephalomyelitis (EAE). This finding has led to a new window of therapeutic opportunity in MS. Monoclonal antibodies to VLA-4 abrogate the development of EAE in sensitised animals and may actually reverse its clinical and pathological findings in manifest disease. Natalizumab, one such monoclonal antibody, which is administered intravenously, has been found to be a promising agent in the treatment of MS. Although single doses produced no improvement in the speed or quality of recovery from acute exacerbations of MS in a phase II trial, long-term administration (in phase II and phase III trials) have produced significant benefits with results showing both a marked reduction in the risk of new magnetic resonance imaging lesions and a significant reduction in the risk of exacerbations within 2 months of the initiation of therapy. Phase III double blinded controlled trials have provided additional evidence of safety and a favourable impact on exacerbation rates over the 1 year of administration. Unfortunately, the success of natalizumab has been curtailed by three cases of progressive multifocal leukoencephalopathy, which have prompted the manufacturer to voluntary withdraw the drug from the market. An independent review board is currently investigating the safety of the drug to determine whether it should return to the market. The demonstration that selective modulation (blocking) of the adhesion molecule VLA-4 by natalizumab in MS, resembling that observed in experimental disease, represents a major advance in rational therapy. PMID- 16268664 TI - Advances in drug treatments for children and adolescents with autism and other pervasive developmental disorders. AB - Autism is a disorder characterised by abnormalities in language and social development, and repetitive behaviours. Antipsychotics, including haloperidol and risperidone, are the most widely studied drugs for reducing symptoms in children and adolescents with autism. When administered at relatively low dosages, antipsychotics have been shown to reduce repetitive behaviours (stereotypies) and social withdrawal, as well as a number of related symptoms, such as hyperactivity, aggression, self-abusive behaviour, temper tantrums, lability of mood and irritability. Adverse effects of antipsychotics include sedation, dizziness, increased appetite, weight gain, changes in the electrocardiogram parameters, drooling, hyperprolactinemia and a risk of drug-related dyskinesias. Other agents have been less well studied for the treatment of autism, but there are suggestive data regarding their safety and efficacy. Of these agents, a number have been investigated, based on theories about the aetiology of autism, including SSRIs and naltrexone, although the efficacy of these agents has been limited. Stimulant drugs have been shown to reduce hyperactivity and improve focus, but they may cause behavioural worsening, weight loss and stereotypies de novo. Secretin is a treatment that has received much media attention after reports of efficacy from small open studies, but all controlled studies have failed to show any benefit. In autism, alternative treatments have also been used, but none have shown benefit in well-designed studies. PMID- 16268665 TI - Head-to-head comparison of the costs of atypical antipsychotics: a systematic review. AB - In many countries, prescribing guidelines recommend the use of atypical or second generation antipsychotics (SGAs) in the first-line treatment of individuals with newly diagnosed schizophrenia. This recommendation has increased the utilisation of these agents and, consequently, produced a progressive increase in the proportion of total direct costs in schizophrenia accounted for by drug therapy. In this still-evolving context of care, it becomes relevant to critically investigate the literature base on the relative cost effectiveness of each SGA in comparison with the others, the purpose being to ascertain whether the data reveal any one agent to be truly more cost effective than the others.A systematic search of economic evaluations comparing two or more SGAs yielded 19 studies meeting the inclusion criteria. Of these, 11 were retrospective database or chart review analyses, six were observational prospective or mirror-image studies, and two were randomised clinical trials. Olanzapine and risperidone were compared in 16 studies, two studies compared clozapine, olanzapine and risperidone, and one compared clozapine and risperidone. While experimental studies indicated an absence of differences among the SGAs in terms of total expenditure, database analyses found contrasting evidence. These latter studies, although susceptible to bias and confounding, should theoretically provide an added dimension, in that they are based on observations from 'real world' practice. However, there were too many potential threats to the validity of these analyses to draw a firm conclusion that any one agent is truly more cost effective than the others. In this uncertain situation, clinicians and policy makers should be aware that indirect evidence from independent randomised controlled trials comparing individual SGAs with haloperidol suggested similar cost effectiveness. As healthcare providers in different settings are ultimately the ones who pay for new innovations, it seems appropriate that they commission research into the cost effectiveness of SGAs. PMID- 16268666 TI - The use of triptans in the management of menstrual migraine. AB - Many women experience headaches, including migraine, in association with their menstrual cycles. Although definitions vary, menstrual migraine generally refers to migraine without aura that occurs within several days prior to and several days after the onset of menses. Although menstrual migraine has been reported to be more difficult to treat than other types of migraines, there is no evidence from controlled clinical trials to support this assertion. Thus, the pharmacological treatment of menstrual migraine should be similar to that of other types of migraines, except with respect to the use of hormonal manipulations to treat menstrual migraine. Serotonin 5-HT(1B/1D) receptor agonists (triptans) are effective for the acute treatment of both menstrual and non-menstrual migraines. When used as acute therapy, a triptan should be administered early, when the headache is still mild in severity. Ideally, an acute therapy will provide rapid and complete pain relief with no disability. Some patients may require preventive therapy for menstrual migraine based on suboptimal response to an adequate trial of acute therapy. Patient diaries that record headache onset, relationship to the menstrual cycle and treatment response through three complete cycles will allow accurate prediction of the onset of menstrual migraine; this information is also needed to make decisions about timing of intermittent preventive therapy. The goals of intermittent preventive therapy are to reduce the frequency, duration and intensity of menstrual migraine attacks. Clinical studies show that triptans are effective when used as either acute therapy or as intermittent preventive therapy for menstrual migraine. Sumatriptan and zolmitriptan have been evaluated in prospective, randomised, controlled trials for acute treatment. Retrospective analyses and open-label studies also support the use of other triptans as acute therapy. In addition, sumatriptan, frovatriptan, naratriptan and zolmitriptan have been evaluated as intermittent preventive therapy in prospective studies. Thus, data from clinical studies indicate that triptans are effective for the treatment of menstrual migraine. PMID- 16268667 TI - Rotigotine: in Parkinson's disease. AB - Rotigotine is a nonergolinic dopamine D3/D2/D1 receptor agonist delivered via a transdermal system and has been evaluated for the treatment of idiopathic Parkinson's disease. Patients with early Parkinson's disease receiving rotigotine monotherapy experienced significantly greater improvements in parkinsonian symptoms (as measured by Unified Parkinson's Disease Rating Scale scores) compared to placebo in two large, well designed clinical trials. Significant beneficial effects versus placebo were observed with the 30 and 40 cm2 rotigotine patches in both trials.black triangle Patients with advanced Parkinson's disease receiving rotigotine as adjunctive therapy with levodopa experienced clinically significant reductions from baseline in 'off' time in two well designed clinical trials. In one trial, a large placebo effect was observed, therefore, there was no significant difference between placebo and active treatment (20, 40 and 60 cm2) for this primary efficacy variable, However, a recent study found a significant (p < or = 0.003) reduction in 'off' time in rotigotine 40 and 60 cm2 recipients versus that in the placebo group. Rotigotine was generally well tolerated in clinical trials as both monotherapy and when administered with levodopa; adverse events were generally mild or moderate in severity. PMID- 16268668 TI - Rotigotine: a viewpoint by Peter LeWitt. PMID- 16268669 TI - Rotigotine: a viewpoint by Tomislav Babic. PMID- 16268670 TI - Size-dependent structures of NanI+n-1 cluster ions with a methanol adsorbate: a combined study by photodissociation spectroscopy and density-functional theory calculation. AB - Methanol adsorption sites on NanI+n-1 ions were investigated. Photoexcitation to charge-transfer states of NanI+n-1 (methanol) predominantly produces two fragment ions: Nan-1I+n-2 (methanol) (neutral NaI loss) and Nan-1I+n-2(neutral NaI and methanol loss), without forming NanI+n-1 (methanol loss). The relative intensities of these fragments are correlated with the geometries and binding energies. PMID- 16268671 TI - Electronic excited-state mixing in NeCl2. AB - Ab initio calculations that explicitly include spin-orbit interactions are reported for the NeCl2 system of electronic states. A surprising curve crossing is observed for the C2v, T-shaped geometry. Away from the C2v geometry, the states mix, as expected. On the basis of these new results we propose a new mechanism for electronic energy transfer from highly vibrationally excited levels of the B electronic state of the chlorine molecule. It is proposed that as long as vibrational predissociation of NeCl2 proceeds by direct coupling of the initial state to the continuum states the Ne atom does not sample geometries that efficiently quench the Cl2 B electronic state. However, when the vibrational dynamics changes to the intramolecular vibrational relaxation regime the Ne atom becomes quite effective at coupling the Cl2 B3Pi0u+ state with a 3Pi2g state. PMID- 16268672 TI - Exchange-correlation functional with broad accuracy for metallic and nonmetallic compounds, kinetics, and noncovalent interactions. AB - By incorporating kinetic-energy density in a balanced way in the exchange and correlational functionals and removing self-correlation effects, we have designed a density functional that is broadly applicable to organometallic, inorganometallic, and nonmetallic bonding, thermochemistry, thermochemical kinetics, and noncovalent interactions as well as satisfying the uniform electron gas limit. The average error is reduced by a factor of 1.3 compared with the best previously available functionals, but even more significantly, we find a functional that has a high accuracy for all four categories of interaction. PMID- 16268673 TI - Oxygen adsorption at anionic free and supported Au clusters. AB - The structure, stability, and O2 adsorption properties of anionic Au(n) (n=1-11) clusters either free or supported at defected MgO100 surfaces are investigated using density-functional theory. O2 adsorption is strong whenever unpaired electrons are present, except for at some small, supported, planar, high-band-gap clusters. These clusters have the unpaired electrons pinned by the Madelung potential of the support. Larger clusters (starting at Au7-Au8) become three dimensional and metallic. This ensures that while one cluster orbital is pinned to the defect, another orbital at comparable energy can undergo depletion, thus binding O2 with charge transfer. PMID- 16268674 TI - Energy relaxation versus spectral diffusion of the OH-stretching vibration of HOD in liquid-to-supercritical deuterated water. AB - The dynamics of vibrational energy relaxation (VER) of the OH-stretching vibration of HOD in liquid-to-supercritical heavy water is studied as a function of temperature and solvent density by femtosecond mid-infrared spectroscopy. Using the dielectric constant of the fluid both, the OH-stretching absorption frequency and the VER rate, can be correlated phenomenologically with the average hydrogen-bond connectivity within the random D2O network. This correlation enables the identification of thermodynamic conditions under which spectral diffusion due to hydrogen-bond breakage/formation is much faster than VER. PMID- 16268675 TI - Molecular dynamics in the isothermal-isobaric ensemble: the requirement of a "shell" molecule. I. Theory and phase-space analysis. AB - Current constant pressure molecular-dynamics (MD) algorithms are not consistent with the recent reformulation of the isothermal-isobaric (NpT) ensemble. The NpT ensemble partition function requires the use of a "shell" molecule to identify uniquely the volume of the system, thereby avoiding the redundant counting of configurations [e.g., G. J. M. Koper and H. Reiss, J. Phys. Chem. 100, 422 (1996); D. S. Corti, Phys. Rev. E, 64, 016128 (2001)]. So far, only the NpT Monte Carlo method has been updated to allow the system volume to be defined by a shell particle [D. S. Corti, Mol. Phys. 100, 1887 (2002)]. A shell particle has yet to be incorporated into MD simulations. The proper modification of the NpT MD algorithm is therefore the subject of this paper. Unlike Andersen's method [H. C. Andersen, J. Chem. Phys. 72, 2384 (1980)] where a piston of unknown mass serves to control the response time of volume fluctuations, the newly proposed equations of motion impose a constant external pressure via the introduction of a shell particle of known mass. Hence, the system itself sets the time scales for pressure and volume fluctuations. The new algorithm is subject to a number of fundamentally rigorous tests to ensure that the equations of motion sample phase space correctly. We also show that the Hoover NpT algorithm [W. G. Hoover, Phys. Rev. A. 31, 1695 (1985); 34, 2499 (1986)] does sample phase correctly, but only when periodic boundary conditions are employed. PMID- 16268676 TI - Molecular dynamics in the isothermal-isobaric ensemble: the requirement of a "shell" molecule. II. Simulation results. AB - The results of a series of constant pressure and temperature molecular-dynamics (MD) simulation studies based on the rigorous shell particle formulation of the isothermal-isobaric (NpT) ensemble are presented. These MD simulations validate the newly proposed constant pressure equations of motion in which a "shell" particle is used to define uniquely the volume of the system [M. J. Uline and D. S. Corti, J. Chem. Phys. (to be published), preceding paper]. Ensemble averages obtained with the new MD NpT algorithm match the ensemble averages obtained using the previously derived shell particle Monte Carlo NpT method [D. S. Corti, Mol. Phys. 100, 1887 (2002)]. In addition, we also verify that the Hoover NpT MD algorithm [W. G. Hoover, Phys. Rev. A 31, 1695 (1985); 34, 2499 (1986)] generates the correct ensemble averages, though only when periodic boundary conditions are employed. The extension of the shell particle MD algorithm to multicomponent systems is also discussed, in which we show for equilibrium properties that the identity of the shell particle is completely arbitrary when periodic boundary conditions are applied. Self-diffusion coefficients determined with the shell particle equations of motion are also identical to those obtained in other ensembles. Finally, since the mass of the shell particle is known, the system itself, and not a piston of arbitrary mass, controls the time scales for internal pressure and volume fluctuations. We therefore consider the effects of the shell particle on the dynamics of the system. Overall, the shell particle MD algorithm is an effective simulation method for studying systems exposed to a constant external pressure and may provide an advantage over other existing constant pressure approaches when developing nonequilibrium MD methods. PMID- 16268677 TI - Direct evaluation of multicomponent phase equilibria using flat-histogram methods. AB - We present a method for directly locating density-driven phase transitions in multicomponent systems. Phase coexistence conditions are determined through manipulation of a total density probability distribution evaluated over a density range that includes both coexisting phases. Saturation quantities are determined through appropriate averaging of density-dependent mean values of a given property of interest. We discuss how to implement the method in both the grand canonical and isothermal-isobaric semigrand ensembles. Calculations can be conducted using any of the recently introduced flat-histogram techniques. Here, we combine the general algorithm with a transition-matrix approach to produce an efficient self-adaptive technique for determining multicomponent phase equilibrium properties. To assess the performance of the new method, we generate phase diagrams for a number of binary and ternary Lennard-Jones mixtures. PMID- 16268678 TI - Efficient treatment of the Hartree interaction in the relativistic Kohn-Sham problem. AB - We elaborate the two-component Douglas-Kroll reduction of the Dirac-Kohn-Sham problem of relativistic density-functional theory as introduced by Matveev and Rosch [J. Chem. Phys. 118, 3997 (2003)]. That method retains corrections to the Coulomb self-interaction (or Hartree) term of the energy functional that are due to the picture change. Using analytic expressions for the matrix elements, one is able to abandon the resolution of the identity approach for a crucial step of the relativistic transformation. Thus, a major source of uncertainties of the method is eliminated because basis sets no longer have to be extended by functions of higher angular momentum, previously required to ensure kinetic balance. This approach also relies on the electron charge-density fitting scheme via an auxiliary basis set. An efficient approximate implementation results if one restricts the relativistic transformation to the spherically symmetric atom centered auxiliary functions. It provides accurate results while simplifying greatly the expressions for the matrix elements of the relativistically transformed operators and significantly reducing the computational effort. We demonstrate the performance of the method for the fine structure of one-electron levels of the Hg atom, the g-tensor shifts of NO2, and the properties of the diatomic molecules Bi2, Pb2, PbO, and TlH. PMID- 16268679 TI - On the accuracy of one-component pseudopotential spin-orbit calculations. AB - Improvements on current one-component extraction procedures of spin-orbit pseudopotentials are investigated for high accuracy computation of spin-orbit coupling energies. By means of the perturbation-theory formalism we first show that spin-orbit pseudopotentials, extracted at the one-component self-consistent field level from a reference all-electron Dirac-Coulomb or Dirac-Coulomb-Breit calculation, include valence spin-orbit polarization and relaxation effects. As a consequence the use of these pseudopotentials in uncontracted spin-orbit configuration interaction (CI) with singles from the reference ground-state configuration gives rise to double counting of these spin-orbit effects. Two new methods that avoid such double counting have been investigated. The first, so called "explicit" method, calculates explicitly, by means of a four-component spin-orbit CI, the double-counted spin-orbit effects and removes them from the pseudopotentials. Due to the nonadditivity of the core and valence spin-orbit effects as well as the so-called "pseudovariational collapse," this method is shown to be cumbersome. In the second "implicit" method the spin-orbit pseudopotential is extracted at the spin-orbit polarized and relaxed level by means of a single-excitation spin-orbit CI calculation. Atomic tests on iodine demonstrate the ability of the latter method to solve the double-counting problem. PMID- 16268680 TI - New constraints upon the electron-electron repulsion energy functional of the one electron reduced density matrix. AB - Three strict constraints upon the electron-electron repulsion energy functional of the one-electron reduced density matrix (the 1-matrix) are obtained by combining its invariance and stationary properties with the extended Koopmans' theorem. The constraints relate the quantities derived from the functional pertaining to an N-electron system with those of its (N-1)-electron congener. Together with the N-representability requirement for the 1-matrix of the congener, identities involving the electron-electron repulsion energies of the two systems and their derivatives with respect to the 1-matrices seriously narrow down the choices for potential approximate density-matrix functionals. This fact is well illustrated in the case of two-electron systems, where the validity of the new constraints is confirmed and found to originate from a nontrivial cancellation among different terms. Thus, the constraints provide a new tool for the construction and testing of new functionals that complements the previously known conditions such as the reproduction of the homogeneous gas energies and momentum distributions, convexity, and the N-representability of the associated 2 matrices. PMID- 16268681 TI - Fast evaluation of polarizable forces. AB - Polarizability is considered to be the single most significant development in the next generation of force fields for biomolecular simulations. However, the self consistent computation of induced atomic dipoles in a polarizable force field is expensive due to the cost of solving a large dense linear system at each step of a simulation. This article introduces methods that reduce the cost of computing the electrostatic energy and force of a polarizable model from about 7.5 times the cost of computing those of a nonpolarizable model to less than twice the cost. This is probably sufficient for the routine use of polarizable forces in biomolecular simulations. The reduction in computing time is achieved by an efficient implementation of the particle-mesh Ewald method, an accurate and robust predictor based on least-squares fitting, and non-stationary iterative methods whose fast convergence is accelerated by a simple preconditioner. Furthermore, with these methods, the self-consistent approach with a larger timestep is shown to be faster than the extended Lagrangian approach. The use of dipole moments from previous timesteps to calculate an accurate initial guess for iterative methods leads to an energy drift, which can be made acceptably small. The use of a zero initial guess does not lead to perceptible energy drift if a reasonably strict convergence criterion for the iteration is imposed. PMID- 16268682 TI - Improvement of semiempirical response properties with charge-dependent response density. AB - The present work outlines a new method for treatment of charge-dependent polarizability in semiempirical quantum models for use in combined quantum mechanical/molecular mechanical simulations of biological reactions. The method addresses a major shortcoming in the performance of conventional semiempirical models for these simulations that is tied to the use of a localized minimal atomic-orbital basis set. The present approach has the advantages that it uses a density basis that retains a set of linear-response equations, does not increase the atomic-orbital basis, and avoids the problem of artificial charge transfer and scaling of the polarizability seen in related models that allow atomic charges to fluctuate. The model introduces four new atom-based parameters and has been tested with the modified neglect of differential overlap d-orbital Hamiltonian against 1132 molecules and ions and shown to decrease the dipole moment and polarizability errors by factors of 2 and 10, respectively, with respect to density-functional results. The method performs impressively for a variety of charge states (from 2+ to 2-), and offers a potentially powerful extension in the design of next generation semiempirical quantum models for accurate simulations of highly charged biological reactions. PMID- 16268683 TI - Coarse-grained free-energy-functional treatment of quasistatic multiscale processes in heterogeneous materials. AB - A new treatment of quasistatic (reversible) multiscale processes in heterogeneous materials at nonzero temperature is presented. The system is coarse grained by means of a finite-element mesh. The coarse-grained free-energy functional (of the positions of the nodes of the mesh) appropriate to the thermodynamic-state variables controlled in the relevant process is minimized. Tests of the new procedure on a Lennard-Jonesium crystal yield thermomechanical properties in good agreement with the "exact" atomistic results. PMID- 16268684 TI - About the calculation of exchange coupling constants using density-functional theory: the role of the self-interaction error. AB - The effect of the correction of the self-interaction error on the calculation of exchange coupling constants with methods based on density-functional theory has been tested in simple model systems. The inclusion of the self-interaction correction cancels the nondynamical correlation energy contributions simulated by the commonly used functionals. Hence, such correction should be important in the accurate determination of exchange coupling constants. We have also tested several recent functionals to calculate exchange coupling constants in transition metal complexes, such as meta-GGA functionals or new formulations of hybrid functionals. The influence of the basis set and of the use of pseudopotentials on the calculated J values has also been evaluated for a Fe(III) dinuclear complex in which the paramagnetic centers bear several unpaired electrons. PMID- 16268685 TI - A theoretical study of molecular conduction. II. A Hartree-Fock approach to transmission probability. AB - In this paper, we discuss molecular conductivity based on Green's function methods. In our calculations, we adopted the self-energy formalism to accommodate semi-infinite electrodes connected to a molecule, and the self-energy was obtained from the surface Green's function of the electrodes. We adopted the formalism of the surface Green's function derived by Sanvito et al. [Phys. Rev. B 59, 11936 (1999)] and Krstic et al. [Phys. Rev. B 66, 205319 (2002)], and although their formalisms for the surface Green's function were different, we were able to demonstrate that these formalisms are mathematically identical. We analyzed the electron transmission probability by using the spectrum expression of Green's function, instead of using the inverse matrix of the effective Hamiltonian that includes an isolated molecule and the electrodes. Finally, we calculated the transmission probability of benzenedithiol based on the Hartree Fock method and analyzed the disappearance of the transmission probability due to the orbital interference. PMID- 16268686 TI - Efficient method for the calculation of time- and frequency-resolved four-wave mixing signals and its application to photon-echo spectroscopy. AB - An efficient method has been developed for the calculation of third-order time- and frequency-resolved optical signals. To obtain the general four-wave mixing signal, seven auxiliary density matrices have to be propagated in time. For the special cases of two-pulse photon-echo and transient-grating signals, two or three density matrices, respectively, are required. The method is limited to weak laser fields (it is thus valid within the third-order perturbation theory) but allows for any pulse durations and automatically accounts for pulse-overlap effects. To illustrate the method, we present the explicit derivation of the three-pulse photon-echo signal. Any other third-order optical signal can be calculated in the same manner. As an example, two- and three-pulse photon-echo and transient-grating signals for a weakly damped displaced harmonic oscillator have been calculated. PMID- 16268687 TI - Theoretical studies on magnetic circular dichroism by the finite perturbation method with relativistic corrections. AB - A theoretical method for calculating magnetic circular dichroism (MCD) of molecules is presented. We examined the numerical accuracy and the stability of the finite perturbation (FP) method and the sum-over-state (SOS) perturbation method. The relativistic effects are shown to be important for the MCD spectra of molecules containing heavy elements. Calculations using the FP and the SOS methods were carried out for ethylene, para- and ortho-benzoquinone, showing that the FP method is superior to the SOS method, as expected. The relativistic effect was examined using the second-order Douglas-Kroll Hamiltonians for the halogen molecules F2, Cl2, Br2, and I2. The Faraday terms of I2 and Br2 were strongly affected by the relativistic effects, while the effect was negligible for Cl2 and F2. PMID- 16268688 TI - A regularized and renormalized electrostatic coupling Hamiltonian for hybrid quantum-mechanical-molecular-mechanical calculations. AB - We describe a regularized and renormalized electrostatic coupling Hamiltonian for hybrid quantum-mechanical (QM)-molecular-mechanical (MM) calculations. To remedy the nonphysical QM/MM Coulomb interaction at short distances arising from a point electrostatic potential (ESP) charge of the MM atom and also to accommodate the effect of polarized MM atom in the coupling Hamiltonian, we propose a partial wave expansion of the ESP charge and describe the effect of a s-wave expansion, extended over the covalent radius r(c), of the MM atom. The resulting potential describes that, at short distances, large scale cancellation of Coulomb interaction arises intrinsically from the localized expansion of the MM point charge and the potential self-consistently reduces to 1r(c) at zero distance providing a renormalization to the Coulomb energy near interatomic separations. Employing this renormalized Hamiltonian, we developed an interface between the Car-Parrinello molecular-dynamics program and the classical molecular-dynamics simulation program Groningen machine for chemical simulations. With this hybrid code we performed QM/MM calculations on water dimer, imidazole carbon monoxide (CO) complex, and imidazole-heme-CO complex with CO interacting with another imidazole. The QM/MM results are in excellent agreement with experimental data for the geometry of these complexes and other computational data found in literature. PMID- 16268689 TI - On the origins of approximations for stochastic chemical kinetics. AB - This paper considers the derivation of approximations for stochastic chemical kinetics governed by the discrete master equation. Here, the concepts of (1) partitioning on the basis of fast and slow reactions as opposed to fast and slow species and (2) conditional probability densities are used to derive approximate, partitioned master equations, which are Markovian in nature, from the original master equation. Under different conditions dictated by relaxation time arguments, such approximations give rise to both the equilibrium and hybrid (deterministic or Langevin equations coupled with discrete stochastic simulation) approximations previously reported. In addition, the derivation points out several weaknesses in previous justifications of both the hybrid and equilibrium systems and demonstrates the connection between the original and approximate master equations. Two simple examples illustrate situations in which these two approximate methods are applicable and demonstrate the two methods' efficiencies. PMID- 16268690 TI - Efficient exact exchange approximations in density-functional theory. AB - Two approaches to approximate the Slater potential component of local exact exchange of density-functional theory are investigated. The first approach employs density fitting of the electrostatic potential integrals over two occupied orbitals and the other approach approximates the "exact" Slater potential with the potential derived from the Becke-Roussel [Phys. Rev. A. 39, 3761 (1989)] model of the exchange hole. In both cases significant time savings can be achieved for larger systems compared to the calculation of the numerical Slater potential. It is then analyzed how well the orbitals obtained from the various total exchange potentials reproduce Hartree-Fock energies and molecular properties. A large range of atoms and small molecules has been utilized, including the three DNA bases adenine, thymine, and cytosine. PMID- 16268691 TI - Accidental vibrational degeneracy in vibrational excited states observed with ultrafast two-dimensional IR vibrational echo spectroscopy. AB - The coupling between the OD stretch v=2 level and benzene-ring modes in 2 methoxyphenol-OD (hydroxyl H replaced by D) is observed with ultrafast two dimensional (2D) IR vibrational echo spectroscopy. Because of this coupling, the 1-2 transition peak in the 2D spectrum is split into a doublet with peaks of approximately equal amplitudes. Several molecules and solvents were used to study this phenomenon. Near-IR (NIR) spectroscopy measurements and density-functional theory calculations (B3LYP6-31+G(d,p) level) were also applied. Experimental results and calculations show that the OD stretch 1-2 transition is coupled to a combination band related to the benzene-ring motions. A simple quantum-mechanical model indicates that the combination band has a frequency of 5172 and 5176.5 cm( 1) in CCl4 and hexane, respectively. The transition between this combination band and the ground state is too weak to detect by NIR. The transition between this band and the OD stretch first excited state is also so weak that most of the intensity of the doublet comes from the oscillator strength produced by coupling to the OD stretch. The model gives the coupling strengths as 6.5 and 7 cm(-1) in CCl4 and hexane, respectively. PMID- 16268692 TI - Ab initio study of the reaction of H2 with an AuPt3 cluster. AB - The study of the interaction of a pyramidal tetramer of AuPt3 with H2 is carried out by means of Hartree-Fock self-consistent field (SCF) calculations using relativistic effective core potentials and multiconfigurational SCF plus multireference variational and perturbational on second-order Moller-Plesset configuration interaction calculations. The AuPt3-H2 interaction was carried out in C(s) symmetry. The three lowest electronic states X 2A", A 2A', and a 4A' of the bare cluster were considered in order to study this interaction. The AuPt3+H2 reaction by a Pt vertex shows that AuPt3 cluster in the three lowest-lying electronic states can spontaneously capture and dissociate the H2 molecule. While, by the AuPt2 face side, the AuPt3 cluster only in the A 2A' electronic state can capture and dissociate the H2 molecule after surmounting a small energy barrier. For the Au vertex, this cluster in the three electronic states can also spontaneously capture and dissociate the H2 molecule. On the other hand, by the Pt3 face side, the AuPt3 cluster is able to capture and dissociate the H2 molecule after surmounting energy barriers, where the AuPt3 (X 2A" and 4A'-H2 adsorption are slightly activated. PMID- 16268693 TI - Structures and stability of medium-sized silicon clusters. III. Reexamination of motif transition in growth pattern from Si15 to Si20. AB - It has been established from experiments that stable medium-sized ionic clusters Si15-Si20 are prolate in shape. Density-functional theories (DFTs) also predict that nearly all low-lying neutral clusters in this size range are prolate in shape. Moreover, most of them are built onto two generic structural motifs, either the tricapped-trigonal-prism (TTP) Si9 motif or the six/six Si6Si6 (sixfold-puckered hexagonal ring Si6 plus six-atom tetragonal bipyramid Si6) motif. However, it appears that the exact location of the TTP-to-six/six motif transition is dependent on the functional (e.g., PBE or BLYP) used in the DFT calculations. Here, we present total-energy calculations for two series of clusters (one series containing six/six motif and the other containing the TTP motif) in the size range of Si16-Si20. The calculations were based on all electron DFT methods with a medium [6-311G (2d)] and a large (cc-pVTZ) basis sets, as well as coupled-cluster single and double substitutions (including triple excitations) [CCSD(T)] method with a modest (cc-pVDZ) basis set. In the DFT calculations, two popular hybrid density functionals, the B3LYP and PBE1PBE, were selected. It is found that the B3LYP total-energy calculations slightly favor the six/six motif, whereas the PBE1PBE calculations slightly favor the TTP motif. The CCSD(T) total-energy calculations, however, show that isomers based on the six/six motif are energetically slightly favorable in the size range of Si16 Si20. Hence, the TTP-to-six/six motif transition is more likely to occur at Si16. PMID- 16268694 TI - Water-ketones hydrogen bonding: the rotational spectrum of cyclobutanone-water. AB - The hydrogen-bonded complex cyclobutanone-water has been studied by Fourier transform molecular-beam microwave spectroscopy in the frequency range of 6-18.5 GHz. The rotational spectra of ten isotopomers have been assigned and measured. Five of them have been obtained from different isotopic species (or configurations) of water (H2O, D2O, DOH, HOD, and H2 18O). The remaining five correspond to the four singly substituted 13C and to the 18O species of cyclobutanone, observed in natural abundance. For all species the inertial defect is in the range from -10.44 to -10.50 uA2, showing that the cyclobutanone frame is effectively planar and that the water molecule is coplanar to this frame. The hydrogen bond, almost linear, is formed between a water proton and one of the lone pairs of the cyclobutanone oxygen. PMID- 16268695 TI - One-color two-photon mass-analyzed threshold ionization spectroscopy of ethyl bromide through a dissociative intermediate state. AB - Mass-analyzed threshold ionization (MATI) spectra of ethyl bromide were obtained using one-color two-photon ionization through a dissociative intermediate state. Accurate values for the adiabatic ionization energy have been obtained, 83099+/-5 and 85454+/-5 cm(-1) for the X1 2E and X2 2E states of the ethyl bromide cation, respectively, giving a splitting of 2355+/-10 cm(-1). Compared with conventional photoelectron data, the two-photon MATI spectrum exhibited a more extensive vibrational structure with a higher resolution, mainly containing the modes involving the dissociation coordinate. The observed modes were analyzed and discussed in terms of wave packet evolving on the potential-energy surface of the dissociative state. PMID- 16268696 TI - Li3-O-Li3 molecule: a metal-nonmetal-metal sandwichlike compound with a distending electron cloud. AB - The D3d and D2d isomers of the Li3-O-Li3 molecule are metal-nonmetal-metal sandwichlike structures that contain two Li3 superalkali atoms. Their geometries and the real frequencies are obtained at the CCSD(T)/aug-cc-pVDZ level. They are different from the traditional types of the nonmetal-metal-nonmetal sandwich compounds. The natural bond orbital calculation and the topological property nabla2rho(r) calculation indicate that they are typical ionic compounds. In two isomers, the O2- anion is sandwiched in between two Li3+ cation rings. However, the different orientations of two Li3+ planes give the D3d isomer its own special characteristics. Under the action of the O2- anion in the center, the valence electrons of the D3d isomer are pushed out from two Li3+ triangle rings. This special interaction causes three phenomena. First, the valence electron clouds are distended. Second, the vertical ionization energy of the D3d isomer is considerably low, 4.39 eV, so that it may also be viewed as a superalkali atom. Third, we find that the D3d isomer owns the out-of-plane aromaticity and the largest negative nucleus-independent chemical shift value (-10.8 ppm) exists at 2.5 A above the center of the Li3+ ring, not at the center of the Li3+ ring like the isolated aromatic Li3+ cation. PMID- 16268697 TI - CS2O+ and CS2O in the gas phase: an experimental and computational study. AB - The CS2O+ ion and CS2O molecule were prepared and structurally characterized by mass spectrometric techniques as isolated species in the gas phase. The theoretical analysis, performed by B3LYP and CCSD(T) computational methods, predicted different CS2O+ isomers, SSCO+, O(CS2)+, SCSO+, SCOS+ and S(COS)+, and structurally related singlet and triplet CS2O. Experiment and theory agree in identifying the obtained CS2O+ ions as a mixture of SCSO+ and SCOS+ isomers. CS2O neutral species, prepared by neutralization-reionization mass spectrometry, were directly characterized as intact, long-lived species with a lifetime tau > or =2 micros. PMID- 16268698 TI - On the control of product yields in the photofragmentation of deuteriumchlorid ions (DCl+). AB - The prospect of controlling the photofragmentation of deuterium chloride ions (DCl+) via strong ultrashort IR laser pulses has been investigated by a numerical solution of coupled Schrodinger equations. The calculations provide evidence that the ratio of product ion yields Cl+ versus D+ can be manipulated by an appropriate choice of laser pulse parameters, in particular, central laser frequency, pulse duration, intensity, and chirp. The analysis of time-dependent populations reveals competition between intra- and interelectronic state excitations, enabling the understanding of quantum control at the molecular level. PMID- 16268699 TI - Mid-infrared characterization of the NH4 +(H2O)n clusters in the neighborhood of the n=20 "magic" number. AB - Vibrational predissociation spectra are reported for size-selected NH4+ (H2O)n clusters (n=5-22) in the 2500-3900 cm(-1) region. We concentrate on the sharp free OH stretching bands to deduce the local H-bonding configurations of water molecules on the cluster surface. As in the spectra of the protonated water clusters, the free OH bands in NH4+ (H2O)n evolve from a quartet at small sizes (n<7), to a doublet around n=9, and then to a single peak at the n=20 magic number cluster, before the doublet re-emerges at larger sizes. This spectral simplification at the magic number cluster mirrors that found earlier in the H+(H2O)n clusters. We characterize the likely structures at play for the n=19 and 20 clusters with electronic structure calculations. The most stable form of the n=20 cluster is predicted to have a surface-solvated NH4+ ion that lies considerably lower in energy than isomers with the NH4+ in the interior. PMID- 16268700 TI - First-principles investigation of finite-temperature behavior in small sodium clusters. AB - A systematic and detailed investigation of the finite-temperature behavior of small sodium clusters, Na(n), in the size range of n=8-50 are carried out. The simulations are performed using density-functional molecular dynamics with ultrasoft pseudopotentials. A number of thermodynamic indicators such as specific heat, caloric curve, root-mean-square bond-length fluctuation, deviation energy, etc., are calculated for each of the clusters. Size dependence of these indicators reveals several interesting features. The smallest clusters with n=8 and 10 do not show any signature of melting transition. With the increase in size, broad peak in the specific heat is developed, which alternately for larger clusters evolves into a sharper one, indicating a solidlike to liquidlike transition. The melting temperatures show an irregular pattern similar to the experimentally observed one for larger clusters [Schmidt et al., Nature (London) 393, 238 (1998)]. The present calculations also reveal a remarkable size sensitive effect in the size range of n=40-55. While Na(40) and Na(55) show well developed peaks in the specific-heat curve, Na(50) cluster exhibits a rather broad peak, indicating a poorly defined melting transition. Such a feature has been experimentally observed for gallium and aluminum clusters [Breaux et al., J. Am. Chem. Soc. 126, 8628 (2004); Breaux et al., Phys. Rev. Lett. 94, 173401 (2005)]. PMID- 16268701 TI - Intermolecular multiple quantum coherences at high magnetic field: the nonlinear regime. AB - Experiments have been carried at magnetic-field strengths of 9.4, 14.1, and 17.6 T to explore the evolution of intermolecular multiple quantum coherences in the nonlinear regime where the system evolves for times that are much greater than the characteristic time of action of the long-range dipolar field, tau(d). The results show the expected Bessel function form of the recorded signal as a function of time of evolution, with evident zeros and sign changes. As expected, the rate of signal evolution increases at higher-field strengths as a result of the increased equilibrium magnetization. A numerical method for calculating the evolution of magnetization under the action of the distant dipolar field, relaxation, and diffusion that is based on Fourier analysis of the magnetization distribution has been applied to the correlated two-dimensional spectroscopy revamped by asymmetric z-gradient echo detection sequence in the nonlinear regime and shown to produce results that are in good agreement with experimental data acquired at different magnetic fields and rates of spatial modulation. Experiments and simulations have also been used to explore the evolution of magnetization in a mixture of two interacting spin species in the nonlinear regime. PMID- 16268702 TI - High-resolution spectroscopy of CoS (X4Deltai): examining 3d transition-metal sulfide bonds. AB - The pure rotational spectrum of CoS, the cobalt sulfide radical, has been measured using direct absorption techniques in the frequency range of 180-540 GHz. This study is the first spectroscopic investigation of any kind of this molecule. CoS was created by reacting cobalt vapor with H2S. Four spin components were identified in the spectra of this species, one of which exhibited lambda doubling, identifying the ground state as 4Deltai. Transitions arising from the lowest spin component of the less abundant Co 35S isotopomer have also been detected, as well as from v=1 and v=2 of the main species. The spectra were readily identified because each spin component exhibited an octet pattern arising from the 59Co spin of I=72. The data were fit using Hund's case (a) Hamiltonian, and rotational, fine-structure, hyperfine, and lambda-doubling constants were determined. The hyperfine parameters support a delta3pi2 electron configuration and are consistent with some orbital overlap between the metal and sulfur atoms. From the rotational constant, the bond length of CoS was calculated to be r0=1.977 985 06(10) A. This bond length is significantly shorter than that of MnS or FeS, in contrast to the bond distances found in the oxide analogs which are all similar in value. These results indicate that the 3d metal sulfides differ somewhat from their oxide counterparts, probably due to the availability of sulfur p orbitals for bonding. PMID- 16268703 TI - State-to-state photodissociation of carbonyl sulfide (nu2=0,1/JlM). II. The effect of initial bending on coherence of S(1D2) polarization. AB - Photodissociation studies using ion imaging are reported, measuring the coherence of the polarization of the S((1)D(2)) fragment from the photolysis of single quantum state-selected carbonyl sulfide (OCS) at 223 and 230 nm. A hexapole state selector focuses a molecular beam of OCS parent molecules in the ground state (nu2=0mid R:JM=10) or in the first excited bending state (nu2=1mid R:JlM=111). At 230 nm photolysis the Im[a1 (1)(parallel, perpendicular)] moment for the fast S(1D2) channel increases by about 50% when the initial OCS parent state changes from the vibrationless ground state to the first excited bending state. No dependence on the initial bending state is found for photolysis at 223 nm. We observe separate rings in the slow channel of the velocity distribution of S(1D2) correlating to single CO(J) rotational states. The additional available energy for photolysis at 223 nm is found to be channeled mostly into the CO(J) rotational motion. An improved value for the OC-S bond energy D0=4.292 eV is reported. PMID- 16268704 TI - Single and double photoionizations of methanal (formaldehyde). AB - Single and double photoionization spectra of formaldehyde have been measured at 40.81 and 48.37 eV photon energy and the spectrum of the doubly charged cation has been interpreted using high-level electronic structure calculations. The adiabatic double-ionization energy is determined as 31.7+/-0.25 eV and the vertical ionization energy is 33 eV. The five lowest excited electronic states are identified and located. The potential-energy surfaces of the accessible states explain the lack of stable H2CO2+ dications and the lack of vibrational structure. The experimental double-ionization spectrum can be decomposed into two distinct contributions, one from direct photoionization and the second from indirect double photoionization by an inner-valence shell Auger effect. PMID- 16268705 TI - Analysis of the rotational spectrum of methylene (CH2) in its vibronic ground state with an Euler expansion of the Hamiltonian. AB - We present an analysis of a global, field-free data set of the methylene radical CH2 in its X 3B1 vibronic ground state by means of a novel Euler expansion of the Hamiltonian. The data set comprises pure rotational transitions up to 2 THz obtained with microwave accuracies of 30-500 kHz as well as nu2 ground-state combination differences and pure rotational data obtained with infrared accuracies of 0.001-0.010 cm(-1). Highly accurate spectroscopic parameters have been determined. These include rotational, spin-spin, spin-rotation, and electron spin-nuclear-spin coupling terms along with several centrifugal distortion corrections. The spectroscopic model has been tested and improved by recording newly three weak DeltaN not equalDeltaJ fine-structure components of the N(KaKc)=2(12)-3(03) and 5(05)-4(14) transitions near 434, 454, and 581 GHz. These lines were rather close to the predictions. Overall weighted root mean squares of 1.28 and 0.83 were achieved for fits in which the Euler expansion was used only for the rotational part of the Hamiltonian or for the rotational and spin-spin terms of the Hamiltonian, respectively. The resulting spectroscopic parameters allow for precise frequency predictions of astrophysically important rotational transitions of methylene. PMID- 16268706 TI - Rotational correlation and dynamic heterogeneity in a kinetically constrained lattice gas. AB - We study the dynamical heterogeneity and glassy dynamics in a kinetically constrained lattice-gas model which has both translational and rotational degrees of freedom. We find that the rotational relaxation time tracks the structural relaxation time as density is increased whereas the translational diffusion constant exhibits a strong decoupling. We investigate distributions of exchange and persistence times for both the rotational and translational degrees of freedom and compare our results on the distributions of rotational exchange times to recent single molecule studies. PMID- 16268707 TI - Structure, thermodynamics, and liquid-vapor equilibrium of ethanol from molecular dynamics simulations using nonadditive interactions. AB - We present a molecular-dynamics simulation study of the bulk and liquid-vapor interfacial properties of ethanol using a polarizable force field based on the fluctuating charge (FQ) formalism, as well as the nonpolarizable CHARMM22 force field. Both models are competitive with respect to the prediction of ambient liquid properties such as liquid density, enthalpy of vaporization, dielectric constant, and self-diffusion constants. The polarizable model predicts an average condensed-phase dipole moment of 2.2 D associated with an induced liquid-phase dipole moment of 0.6 D; though qualitatively in agreement with earlier nonadditive models as well as recent Car-Parinello calculations, the current FQ model underestimates the condensed-phase dipole moment. In terms of liquid structure, both models are in agreement with recent neutron-diffraction results of liquid ethanol structure, although the polarizable model predicts the hydroxyl hydrogen-hydroxyl-hydrogen structure factor in closer agreement with the experimental data. In terms of interfacial properties, both models predict ambient surface tension to within 4% of the experimental value of 22.8 dyncm, while overestimating the surface excess entropy by almost a factor of 2. Both models display the characteristic preferential orientation of interfacial molecules. The polarizable model allows for a monotonic variation of the average molecular dipole moment from the bulk value to that of the vapor phase. Consequently, there is a dramatic difference in the surface potential predicted by the polarizable and nonpolarizable models. The polarizable model estimates a surface potential of -209+/-3 mV, while the nonpolarizable model yields a value of -944+/-10 mV. Finally, based on the vapor-liquid equilibrium simulation data from several temperatures, we estimate the critical properties of both models. As observed with other FQ models for associating fluids (such as water and methanol), and counter to what one would anticipate by modeling more physically the electrostatic response to local environment, the current FQ model underestimates the critical temperature and overestimates the critical density of ethanol; moreover, the FQ model is, in this respect, equivalent to the underlying fixed-charge model. These results further suggest the need to revisit polarizable models in terms of quantitative vapor-liquid equilibrium prediction. PMID- 16268708 TI - Effects of nonpolar solutes on the thermodynamic response functions of aqueous mixtures. AB - We investigate the effect of adding nonpolar solutes at atmospheric pressure on water's temperature of maximum density, isothermal compressibility, and isobaric heat capacity, using a statistical mechanical model of water solutions [H. S. Ashbaugh, T. M. Truskett, and P. G. Debenedetti, J. Chem. Phys. 116, 2907 (2002)]. We find that the temperature of maximum density increases with solute hydrophobicity, as characterized by its size, and decreases with its van der Waals attractive parameter a, in agreement with experiment. We predict similar trends for the addition of solutes on the isothermal compressibility and isobaric heat capacity: solute hydrophobicity causes an upward shift in water's anomalies, whereas dispersive interactions as measured by the solute's van der Waals attractive parameter shift the anomalies to lower temperatures. The locus along which the competing contributions of solute size sigma and interaction strength a to the shift in water's response functions balance each other obeys the scaling relationship sigma6 approximately a. PMID- 16268709 TI - Dynamics of glass-forming liquids. X. Dielectric relaxation of 3-bromopentane as molecular probes in 3-methylpentane. AB - The glass-forming liquids 3-bromopentane (3BP) and 3-methylpentane (3MP) are readily miscible across the entire composition range, although their polarities differ considerably. As noted by Berberian [J. Non-Cryst. Solids 131-133, 48 (1991)], the nearly matching molar volumes makes this binary system appear ideal for probe-sensitized measurements. We have performed a dielectric study of these mixtures in the range of 3BP mole fractions x from 2 x 10(-4) to 0.75. In the limit of low concentrations, x<0.5%, the dielectric loss peak of 3BP is slower by a factor of 2.5 relative to that of 3MP. Additionally, the relaxation behavior of the guest is more exponential than that of the host liquid. We interpret the distinct dynamics of the guest as a result of temporal averaging over the heterogeneous host dynamics, with the exchange time being near the longest structural time constant of the system. PMID- 16268710 TI - On the performance of molecular polarization methods. II. Water and carbon tetrachloride close to a cation. AB - Our initial study on the performance of molecular polarization methods close to a positive point charge [M. Masia, M. Probst, and R. Rey, J. Chem. Phys. 121, 7362 (2004)] is extended to the case in which a molecule interacts with a real cation. Two different methods (point dipoles and shell model) are applied to both the ion and the molecule. The results are tested against high-level ab initio calculations for a molecule (water or carbon tetrachloride) close to Li+, Na+, Mg2+, and Ca2+. The monitored observable is in all cases the dimer electric dipole as a function of the ion-molecule distance for selected molecular orientations. The moderate disagreement previously obtained for point charges at intermediate distances, and attributed to the linearity of current polarization methods (as opposed to the nonlinear effects evident in ab initio calculations), is confirmed for real cations as well. More importantly, it is found that at short separations the phenomenological polarization methods studied here substantially overestimate the dipole moment induced if the ion is described quantum chemically as well, in contrast to the dipole moment induced by a point charge ion, for which they show a better degree of accord with ab initio results. Such behavior can be understood in terms of a decrease of atomic polarizabilities due to the repulsion between electronic charge distributions at contact separations. It is shown that a reparametrization of the Thole method for damping of the electric field, used in conjunction with any polarization scheme, allows to satisfactorily reproduce the dimer dipole at short distances. In contrast with the original approach (developed for intramolecular interactions), the present reparametrization is ion and method dependent, and corresponding parameters are given for each case. PMID- 16268711 TI - A new high-temperature multinuclear-magnetic-resonance probe and the self diffusion of light and heavy water in sub- and supercritical conditions. AB - A high-resolution nuclear-magnetic-resonance probe (500 MHz for 1H) has been developed for multinuclear pulsed-field-gradient spin-echo diffusion measurements at high temperatures up to 400 degrees C. The convection effect on the self diffusion measurement is minimized by achieving the homogeneous temperature distributions of +/-1 and +/-2 degrees C, respectively, at 250 and 400 degrees C. The high temperature homogeneity is attained by using the solid-state heating system composed of a ceramic (AlN) with high thermal conductivity comparable with that of metal aluminium. The self-diffusion coefficients D for light (1H2O) and heavy (2H2O) water are distinguishably measured at subcritical temperatures of 30 350 degrees C with intervals of 10-25 degrees C on the liquid-vapor coexisting curve and at a supercritical temperature of 400 degrees C as a function of water density between 0.071 and 0.251 gcm3. The D value obtained for 1H2O is 10%-20% smaller than those previously reported because of the absence of the convection effect. At 400 degrees C, the D value for 1H2O is increased by a factor of 3.7 as the water density is reduced from 0.251 to 0.071 gcm3. The isotope ratio D(1H2O)D(2H2O) decreases from 1.23 to approximately 1.0 as the temperature increases from 30 to 400 degrees C. The linear hydrodynamic relationship between the self-diffusion coefficient divided by the temperature and the inverse viscosity does not hold. The effective hydrodynamic radius of water is not constant but increases with the temperature elevation in subcritical water. PMID- 16268712 TI - Search for memory effects in methane hydrate: structure of water before hydrate formation and after hydrate decomposition. AB - Neutron diffraction with HD isotope substitution has been used to study the formation and decomposition of the methane clathrate hydrate. Using this atomistic technique coupled with simultaneous gas consumption measurements, we have successfully tracked the formation of the sI methane hydrate from a water/gas mixture and then the subsequent decomposition of the hydrate from initiation to completion. These studies demonstrate that the application of neutron diffraction with simultaneous gas consumption measurements provides a powerful method for studying the clathrate hydrate crystal growth and decomposition. We have also used neutron diffraction to examine the water structure before the hydrate growth and after the hydrate decomposition. From the neutron-scattering curves and the empirical potential structure refinement analysis of the data, we find that there is no significant difference between the structure of water before the hydrate formation and the structure of water after the hydrate decomposition. Nor is there any significant change to the methane hydration shell. These results are discussed in the context of widely held views on the existence of memory effects after the hydrate decomposition. PMID- 16268713 TI - Caging and mosaic length scales in plaquette spin models of glasses. AB - We consider two systems of Ising spins with plaquette interactions. They are simple models of glasses which have dual representations as kinetically constrained systems. These models allow an explicit analysis using the mosaic, or entropic droplet, approach of the random first-order transition theory of the glass transition. We show that the low-temperature states of these systems resemble glassy mosaic states, despite the fact that excitations are localized and that there are no static singularities. By means of finite-size thermodynamics we study a generalized caging effect whereby the system is frozen on short length scales, but free at larger length scales. We find that the freezing length scales obtained from statics coincide with those relevant to dynamic correlations, as expected in the mosaic view. The simple nucleation arguments of the mosaic approach, however, do not give the correct relation between freezing lengths and relaxation times, as they do not capture the transition states for relaxation. We discuss how these results make a connection between the mosaic and the dynamic facilitation views of glass formers. PMID- 16268715 TI - Electrolyte diodes with weak acids and bases. II. Numerical model calculations and experiments. AB - This is the second part of our work dealing with electrolyte diodes with weak acids and bases. In the first part an approximative analytical solution was derived for the steady-state current-voltage characteristic (CVC) of a reverse biased diode (a quasi-one-dimensional gel connecting an acidic and an alkaline reservoir), applying either strong or weak electrolytes. An approximative analytical solution is compared here with a numerical solution free of any approximations and with CVCs measured experimentally with both strong and weak electrolytes. It is shown that the deviations between the numerical and analytical solutions are mostly due to assumptions made for the fixed charge concentration profiles. The concept of optimal analytical solution is introduced which does not use such assumptions and applies only the quasielectroneutrality and quasiequilibrium approximations. It is proven that the slope of the CVC based on the optimum analytical solution can be calculated without the complicated derivation of that solution itself. The calculation of that slope is based on the fact that in the optimum analytical solution all currents are inversely proportional to the length if the boundary conditions are held constant and realizing that in the middle part of the gel the only mobile counterions of the fixed ionized groups are hydrogen ions. In the experimental part the apparatus and the preparation of the gel are described together with the CVCs measured with strong and weak electrolytes. From these CVCs the fixed ion concentration in the middle part of the gel can be determined. That fixed ion concentration is 1.96 x 10(-4)M measured with weak electrolytes and 3.48 x 10(-4)M measured with strong electrolytes. The deviation indicates that the strong base causes some hydrolysis of the gel. Finally, possible applications of weak acid-weak base diodes are discussed. PMID- 16268714 TI - Electrolyte diodes with weak acids and bases. I. Theory and an approximate analytical solution. AB - Until now acid-base diodes and transistors applied strong mineral acids and bases exclusively. In this work properties of electrolyte diodes with weak electrolytes are studied and compared with those of diodes with strong ones to show the advantages of weak acids and bases in these applications. The theoretical model is a one dimensional piece of gel containing fixed ionizable groups and connecting reservoirs of an acid and a base. The electric current flowing through the gel is measured as a function of the applied voltage. The steady-state current-voltage characteristic (CVC) of such a gel looks like that of a diode under these conditions. Results of our theoretical, numerical, and experimental investigations are reported in two parts. In this first, theoretical part governing equations necessary to calculate the steady-state CVC of a reverse biased electrolyte diode are presented together with an approximate analytical solution of this reaction-diffusion-ionic migration problem. The applied approximations are quasielectroneutrality and quasiequilibrium. It is shown that the gel can be divided into an alkaline and an acidic zone separated by a middle weakly acidic region. As a further approximation it is assumed that the ionization of the fixed acidic groups is complete in the alkaline zone and that it is completely suppressed in the acidic one. The general solution given here describes the CVC and the potential and ionic concentration profiles of diodes applying either strong or weak electrolytes. It is proven that previous formulas valid for a strong acid-strong base diode can be regarded as a special case of the more general formulas presented here. PMID- 16268716 TI - Physics of solid and liquid alkali halide surfaces near the melting point. AB - This paper presents a broad theoretical and simulation study of the high temperature behavior of crystalline alkali halide surfaces typified by NaCl(100), of the liquid NaCl surface near freezing, and of the very unusual partial wetting of the solid surface by the melt. Simulations are conducted using two-body rigid ion Born-Mayer-Huggins-Fumi-Tosi (BMHFT) potentials, with full treatment of long range Coulomb forces. After a preliminary check of the description of bulk NaCl provided by these potentials, which seems generally good even at the melting point, we carry out a new investigation of solid and liquid surfaces. Solid NaCl(100) is found in this model to be very anharmonic and yet exceptionally stable when hot. It is predicted by a thermodynamic integration calculation of the surface free energy that NaCl(100) should be a well-ordered, nonmelting surface, metastable even well above the melting point. By contrast, the simulated liquid NaCl surface is found to exhibit large thermal fluctuations and no layering order. In spite of that, it is shown to possess a relatively large surface free energy. The latter is traced to a surface entropy deficit, reflecting some kind of surface short-range order. We show that the surface short range order is most likely caused by the continuous transition of the bulk ionic melt into the vapor, made of NaCl molecules and dimers rather than of single ions. Finally, the solid-liquid interface free energy is derived through Young's equation from direct simulation of partial wetting of NaCl(100) by a liquid droplet. The resulting interface free energy is large, in line with the conspicuous solid-liquid 27% density difference. A partial wetting angle near 50 degrees close to the experimental value of 48 degrees is obtained in the process. It is concluded that three elements, namely, the exceptional anharmonic stability of the solid (100) surface, the molecular short-range order at the liquid surface, and the costly solid-liquid interface, all conspire to cause the anomalously poor wetting of the (100) surface by its own melt in the BMHFT model of NaCl-and most likely also in real alkali halide surfaces. PMID- 16268717 TI - Rotational effects in the dissociative adsorption of H2 on the Pt211 stepped surface. AB - Rotational effects in the dissociative adsorption of H2 on the Pt211 stepped surface have been studied using classical trajectory calculations on a six dimensional, density-functional theory potential-energy surface. Reaction of rotating molecules via an indirect trapping mechanism exhibits an unexpected nonmonotonic dependence on the initial rotational quantum number J. Indirect reaction is first quenched with increasing J but is enhanced again for high J initial states. The quenching is attributed to rotational-to-translational energy transfer, which facilitates escape from the chemisorption wells responsible for molecular trapping. For high J, rotational and translational motions decouple, and the energy transfer is no longer possible, which leads again to trapping. Degeneracy-resolved calculations show that for high initial J, molecules rotating in a "cartwheel" fashion (mJ=0) are more likely to become trapped and react indirectly than "helicoptering" molecules (mJ=J). Experimental confirmation of this finding would lend strong support to the existence of the chemisorption wells that trap molecules prior to reaction. PMID- 16268718 TI - Evidence of overcharging in the complexation between oppositely charged polymers and surfactants. AB - We report on the complexation between charged-neutral block copolymers and oppositely charged surfactants studied by small-angle neutron scattering. Two block copolymers/surfactant systems are investigated, poly(acrylicacid)-b poly(acrylamide) with dodecyltrimethylammonium bromide and poly(trimethylammonium ethylacrylate methylsulfate)-b-poly(acrylamide) with sodium dodecyl sulfate. Two two systems are similar in terms of structure and molecular weight but have different electrostatic charges. The neutron-scattering data have been interpreted in terms of a model that assumes the formation of mixed polymer surfactant aggregates, also called colloidal complexes. These complexes exhibit a core-shell microstructure, where the core is a dense coacervate microphase of micelles surrounded by neutral blocks. Here, we are taking advantage of the fact that the complexation results in finite-size aggregates to shed some light on the complexation mechanisms. In order to analyze quantitatively the neutron data, we develop two different approaches to derive the number of surfactant micelles per polymer in the mixed aggregates and the distributions of aggregation numbers. With these results, we show that the formation of the colloidal complex is in agreement with overcharging predictions. In both systems, the amount of polyelectrolytes needed to build the core-shell colloids always exceeds the number that would be necessary to compensate the charge of the micelles. For the two polymer-surfactant systems investigated, the overcharging ratios are 0.66+/ 0.06 and 0.38+/-0.02. PMID- 16268719 TI - Second-harmonic generation in an optically poled azo-dye/polymer film. AB - The power dependence of the optical poling process of a new azo chromophore in the polymethylmethacrylate matrix at room temperature has been investigated. The existing theory is found to be inconsistent with the experiment. A simple model based on the rate equation is proposed to describe the writing process of the chromophore/polymer system. The measured growth rate and the plateau second harmonic generation intensity as induced by all optical poling are found to be in good agreement with the prediction of the simple model. PMID- 16268720 TI - Rotational diffusion of colloidal particles near confining walls. AB - We study the rotational diffusion of a spherical colloid confined in a narrow channel between parallel plane hard walls. The walls damp translational diffusion much more than rotational diffusion so that there is expected to be little translation-rotation coupling. Using a recent calculation of the nonisotropic rotational mobilities arising from the hydrodynamic interactions with the walls, we set up the rotational Smoluchowski equation for either a particle with a permanent dipole moment or a polarizable particle with axisymmetric polarizabilities subject to an external electric field. Using the Smoluchowski equation dynamics we calculate the time-correlation functions of orientation that are measured in depolarized light scattering for the cases of no external field, external field normal to the walls, and external field parallel to the walls. The decay of correlations is shown to be given by a weighted sum of decaying exponentials and can be characterized by an initial and a mean characteristic decay time. The weights and decay rates of each component and the characteristic decay times are studied numerically for a range of field strengths. The nonisotropic rotational mobilities make these decay times highly sensitive to the distance of the particle from the confining walls. This position dependence can be used as a method of measuring the rotational mobilities or, conversely, the rate of decay of correlations can be used as a probe of particle position between the confining walls. PMID- 16268721 TI - Replica Ornstein-Zernike theory of adsorption in a templated porous material: interaction site systems. AB - Molecular templating offers the possibility of porous materials whose selectivity rivals the molecular recognition observed in nature. The design of templated materials requires a molecular understanding of the templating effect on the material structure and performance. We present here a theoretical description of adsorption in a model templated porous material. Our model material is a quenched, equilibrated mixture of template and matrix molecular species where the template component has been subsequently removed. We propose a set of site-site [i.e., reference interaction site model (RISM)] replica Ornstein-Zernike equations relating the correlation functions of template, matrix, and adsorbing fluid molecules. To test this approach, we focus here on systems interacting via hard-sphere site-site potentials and employ a Percus-Yevick closure. We consider chain and cluster species composed of up to five spheres and observe a range of effects associated with template structure, including higher affinity toward, and enhanced templating by, compact cluster molecules. We assess these effects by grand canonical Monte Carlo simulation and discuss their implication to the design of templated molecular recognition materials. PMID- 16268722 TI - Competition of mesoscales and crossover to theta-point tricriticality in near critical polymer solutions. AB - The approach to asymptotic critical behavior in polymer solutions is governed by a competition between the correlation length of critical fluctuations diverging at the critical point of phase separation and an additional mesoscopic length scale, the radius of gyration. In this paper we present a theory for crossover between two universal regimes: a regime with Ising (fluctuation-induced) asymptotic critical behavior, where the correlation length prevails, and a mean field tricritical regime with theta-point behavior controlled by the mesoscopic polymer chain. The theory yields a universal scaled description of existing experimental phase-equilibria data and is in excellent agreement with our light scattering experiments on polystyrene solutions in cyclohexane with polymer molecular weights ranging from 2 x 10(5) up to 11.4 x 10(6). The experiments demonstrate unambiguously that crossover to theta-point tricriticality is controlled by a competition of the two mesoscales. The critical amplitudes deduced from our experiments depend on the polymer molecular weight as predicted by de Gennes [Phys. Lett. 26A, 313 (1968)]. Experimental evidence for the presence of logarithmic corrections to mean-field tricritical theta-point behavior in the molecular-weight dependence of the critical parameters is also presented. PMID- 16268723 TI - Nonlinear elasticity and yielding of depletion gels. AB - A microscopic activated barrier hopping theory of the viscoelasticity of colloidal glasses and gels has been generalized to treat the nonlinear rheological behavior of particle-polymer suspensions. The quiescent cage constraints and depletion bond strength are quantified using the polymer reference interaction site model theory of structure. External deformation (strain or stress) distorts the confining nonequilibrium free energy and reduces the barrier. The theory is specialized to study a limiting mechanical description of yielding and modulus softening in the absence of thermally induced barrier hopping. The yield stress and strain show a rich functional dependence on colloid volume fraction, polymer concentration, and polymer-colloid size asymmetry ratio. The yield stress collapses onto a master curve as a function of the polymer concentration scaled by its ideal mode-coupling gel boundary value, and sufficiently deep in the gel is of an effective power-law form with a universal exponent. A similar functional and scaling dependence of the yield stress on the volume fraction is found, but the apparent power-law exponent is nonuniversal and linearly correlated with the critical gel volume fraction. Stronger gels are generally, but not always, predicted to be more brittle in the strain mode of deformation. The theoretical calculations appear to be in accord with a broad range of observations. PMID- 16268724 TI - Dynamic yielding, shear thinning, and stress rheology of polymer-particle suspensions and gels. AB - The nonlinear rheological version of our barrier hopping theory for particle polymer suspensions and gels has been employed to study the effect of steady shear and constant stress on the alpha relaxation time, yielding process, viscosity, and non-Newtonian flow curves. The role of particle volume fraction, polymer-particle size asymmetry ratio, and polymer concentration have been systematically explored. The dynamic yield stress decreases in a polymer concentration- and volume-fraction-dependent manner that can be described as apparent power laws with effective exponents that monotonically increase with observation time. Stress- or shear-induced thinning of the viscosity becomes more abrupt with increasing magnitude of the quiescent viscosity. Flow curves show an intermediate shear rate dependence of an effective power-law form, becoming more solidlike with increasing depletion attraction. The influence of polymer concentration, particle volume fraction, and polymer-particle size asymmetry ratio on all properties is controlled to a first approximation by how far the system is from the gelation boundary of ideal mode-coupling theory (MCT). This emphasizes the importance of the MCT nonergodicity transition despite its ultimate destruction by activated barrier hopping processes. Comparison of the theoretical results with limited experimental studies is encouraging. PMID- 16268725 TI - A simple theory of molecular organization in fullerene-containing liquid crystals. AB - Systematic efforts to synthesize fullerene-containing liquid crystals have produced a variety of successful model compounds. We present a simple molecular theory, based on the interconverting shape approach [Vanakaras and Photinos, J. Mater. Chem. 15, 2002 (2005)], that relates the self-organization observed in these systems to their molecular structure. The interactions are modeled by dividing each molecule into a number of submolecular blocks to which specific interactions are assigned. Three types of blocks are introduced, corresponding to fullerene units, mesogenic units, and nonmesogenic linkage units. The blocks are constrained to move on a cubic three-dimensional lattice and molecular flexibility is allowed by retaining a number of representative conformations within the block representation of the molecule. Calculations are presented for a variety of molecular architectures including twin mesogenic branch monoadducts of C60, twin dendromesogenic branch monoadducts, and conical (badminton shuttlecock) multiadducts of C60. The dependence of the phase diagrams on the interaction parameters is explored. In spite of its many simplifications and the minimal molecular modeling used (three types of chemically distinct submolecular blocks with only repulsive interactions), the theory accounts remarkably well for the phase behavior of these systems. PMID- 16268726 TI - Quantum chemical studies on polythiophenes containing heterocyclic substituents: effect of structure on the band gap. AB - Color tuning by the tailoring of substituents at the 3-position of thiophene is very encouraging, and comparative experimental and theoretical studies proved to be powerful in the search for a suitable design for the above. Since the novel polythiophene-based materials substituted with five-membered/six-membered ring containing sulphur and nitrogen at different positions are proven to be potential candidates for electron-transporting hole blocking functions, the structure property relationship of these systems have been focused in the present study. Molecular-orbital calculations are applied to obtain the optimized geometries and band gaps of the thiophene oligomers. An oligomeric approach has been implemented for calculating the band gaps, and the theoretically obtained band gaps for the different model compounds are compared. Density-functional theory B3LYP6-31G* predicted band-gap values are compared with the experimental band gaps obtained from optical-absorption edge. The predicted values show little deviations from experimental band gaps, but the trend in band gap is found to be the same in experimental and theoretical results in most of the cases. Hence, this study illustrates the usefulness of quantum-mechanical calculations in understanding the effects of various structural parameters on optical band gap. PMID- 16268727 TI - Comment on "the effect of density on the properties of short chain fluids" [J. Chem. Phys. 122, 094906 (2005)]. PMID- 16268729 TI - Comment on "hydrophobic effects on partial molar volume" [J. Chem. Phys. 122, 094509 (2005)]. PMID- 16268734 TI - Serotonin and depression: a disconnect between the advertisements and the scientific literature. PMID- 16268735 TI - Comprehensive photochemistry and photophysics of land- and marine-based beta carbolines employing time-resolved emission and flash transient spectroscopy. AB - The photophysical and photochemical behavior of Norharmane (Norh), Harmane (Hara) and Harmine (Hari) and their cations have been examined as a function of the nature of the solvent. Time-resolved emission in nonprotic polar solvents showed fluorescence for all and also phosphorescence for Hari. All emissions were assigned as those of the neutral molecules. Norh and Hari showed fluorescence of both the neutral and the cation in methanol as well as phosphorescence of the neutral while Hari also had fluorescence of the zwitter ion. In ethanol, Norh and Hari displayed fluorescence and phosphorescence of the neutral. The ground-state cations of Norh and Hari exhibited fluorescences of the cation and Hari also had a phosphorescence (cation). The flash transient spectra in nonprotic solvents of all three carbolines had long-lived triplet transients only of the neutral. Triplet and singlet oxygen yields were quite high, 0.31-0.40. Direct excitation of any of the cations gave only the cation triplet. The triplet yields of the cations appear to be low (0.01-0.10 range). Theoretical calculations were done relative to location of triplet states. Some new information will be reported on other naturally occurring differently substituted marine-based beta-carbolines. The impact of all of the foregoing observations on the photosensitizing potential of all compounds is discussed. PMID- 16268736 TI - [Uncertainty on drug effectiveness and safety. Relevance of studies on drug use]. PMID- 16268737 TI - [Assisted electronic prescription: a new opportunity for leadership regarding knowledge management in drug therapy]. PMID- 16268738 TI - [Comparative study of errors in electronic versus manual prescription]. AB - OBJECTIVE: Electronic prescribing is considered a basic measure for the prevention and reduction of medications errors. The goal of this survey was to assess the incidence of errors occurring with electronic versus standard prescription. METHOD: A prospective, sequential, open-label study to assess errors with electronic prescribing as compared to traditional manual prescribing in two public hospitals in Balearic Islands. Errors regarding medication, diet and/or nursing orders were assesses along four process stages: medical prescription, pharmacy transcription/validation, nursing transcription, and dispensation. RESULTS: With manual prescription 1,576 errors/18,539 therapy orders (8.50%) were identified, whereas with electronic prescription 827 errors/18,885 therapy orders (4.38%) were detected, which represents a relative risk reduction by 48% and an absolute risk reduction by 4.12% (p < 0.0001). Pharmacy transcription/validation errors decreased (1.73 vs. 0.13%, p < 0.0001), as did nursing transcription errors (2.54 vs. 0.81%, p < 0.0001) and dispensation errors (2.13 vs. 0.96%, p < 0.0001); however, the number of prescription errors increased (2.10 vs. 2.40%, p = 0.0401). CONCLUSIONS: Electronic prescription is a powerful tool, and one that in this work was shown to decrease medication-, diet , and nursing care-related errors in a highly significant way; however, it should be developed and maintained in order to achieve safety and effectiveness as required by drug usage. PMID- 16268739 TI - [Computer-aided electronic prescribing in Spanish hospitals]. AB - OBJECTIVE: To describe electronic prescribing in a sample of Spanish hospitals and activities derived from its implementation in Hospital Pharmacy Departments, software features and acceptance. METHOD: A survey that included data related to program implantation and performance in hospitals with diverse characteristics was designed. It was posted at the Sociedad Espanola de Farmacia Hospitalaria website for a 6-month period and completed by phone. RESULTS: A total of 47 hospitals answered the survey. 13 (27.7%) had electronic prescribing systems, and of these, 8 of them had more than 75% of beds included in the system. In 15 (31.9%) hospitals that did not have it, its implementation was in project in the near future. Software applications used in this new health technology were not homogeneous, while integration of applications that facilitate Pharmaceutical Care tasks was infrequent. In 61.5% of the hospitals with electronic prescribing, transcription used to be the pharmacist s task. In 92.3% of these hospitals, order validation is currently carried out by pharmacists. The degree of satisfaction with the system was rated as good or very good by 92.3% of pharmacists. CONCLUSIONS: Computer-aided electronic prescribing appears to be a helpful tool for restructuring Department organization and investing time in new activities. A more ambitious and effective approach to clinical activities will therefore be possible. System optimization requires taking into account factors such as worker time investment, since new activities will be developed, along with the need for training courses. PMID- 16268740 TI - [Designing a module for the prevention of hypersensitivity reactions in an assisted electronic prescription system]. AB - OBJECTIVE: To develop a module for the prevention of drug-related allergies to be integrated within the assisted electronic prescription software PRISMA. METHOD: On module design potential sources of medication errors regarding drug allergies were first analyzed, and ideal module characteristics were defined. Then a review of the literature was performed to define "group allergies", and last of all master archives were created, with their required relations being established. RESULTS: A module for the prevention of drug-related allergies in the setting of an assisted electronic prescription software was designed. By interrelating tables listing active ingredients, excipients, chemical structures/functional groups, and "group allergies" prescriptions may be interactively checked, and useful information is provided to the prescribing practitioner--as well as the whole multidisciplinary team--to help him make his/her decisions. PMID- 16268741 TI - [Perioperative anaemia in geriatric patients with hip fracture]. AB - OBJECTIVE: To describe and study the effectiveness of the perioperative anaemia treatment patterns for patients older than 64 with hip fracture. METHOD: Three groups of patients were compared: Group 1: Oral iron or without iron therapy. Group 2: low doses of intravenous iron. Group 3: treated according to a blood saving programme including intravenous iron, alpha epoetin and restrictive transfusional criteria. The homogeneity of gender, age, type of fracture, ASA, preoperative period and perisurgical bleeding affecting drug consumption within the groups was analyzed. The effectiveness of the treatments was determined by means of transfusional rate, postoperative haemoglobin levels, and postoperative length of stay and infection rate. RESULTS: 329 patients were checked. Patients were comparable. Patients included in Group 3 were transfused less than the rest (36.5 of patients in group 3 vs. 52.0% in group 1 and 67.6% in group 3, p = 0.002). Decreases in the infection rate and mean postoperative stay in group 3 were not significant. Haemoglobin levels at 48 hours post surgery were higher in group 1 but haemoglobin levels at the seventh day post surgery were similar for the three groups. CONCLUSIONS: The above mentioned blood saving programme has been observed to be effective in decreasing transfusional requirements without increasing morbidity. However, further prospective studies are needed in order to define the cost-effectiveness of this programme and to determine its role in the reduction of posttransfusional infections and postoperative length of stay. PMID- 16268742 TI - [Effectiveness of heparin versus NaCl 0.9% in central venous catheter flushing. A systematic review]. AB - OBJECTIVE: To determine the effects of intermittent flushing with heparin versus NaCl 0.9% solution on prevention of thrombotic events and infections in central venous catheters. METHOD: A systematic review of controlled trials, randomized or not, in whose intermittent flushing with heparin versus NaCl 0.9% in central venous catheters were compared was carried out. Any publication was excluded because of patients age, country or language. Studies were identified by computerised searches (MEDLINE and CINAHL 1996-2004, EMBASE Drugs and Pharmacology 1991-2004, Cochrane Library up 2005, ISI Web of knowledge 1990 2004), clinical trials metaregister http://www. controlled trials.commrct/mrct_info_es.asp, and scanning references. RESULTS: Only two clinical trials, involving 94 patients, fulfilled inclusion criteria for obstruction. Methodological quality of trials was poor and heterogenicity was also very significant. It made impossible meta-analytical combination of results. No studies comparing heparin vs. NaCl 0.9% solution flushing in catheter-related infection prophylaxis were reported. CONCLUSIONS: Poor evidence supports effectiveness of intermittent heparin flushing vs. NaCl 0.9% solution, although methodological limitations and small sample size make these findings very inconsistent. In infection prophylaxis, there is no data allowing us to conclude if heparin flushing is more effective than NaCl 0.9% solution. More randomized clinical trials are needed. PMID- 16268743 TI - [Leflunomide: assessing teratogenic risk during the first trimester of pregnancy]. AB - OBJECTIVE: To assess the teratogenic risk associated with leflunomide during the first quarter of pregnancy, and to establish guidelines to minimize said risk. METHOD: Literature search using tertiary, secondary, and primary sources related to teratogenicity, including databases (MEDLINE and EMBASE) and specific webs. The information required for assessment, as well as for the establishment of criteria was collected. RESULTS: Leflunomide demonstrated an increased risk of fetal death and teratogenic effects in animals. No major or minor malformation cases have been reported in humans regarding leflunomide, which is classified within category X of fetal risk. A wash-out regimen may possibly reduce the risk for fetal harm. CONCLUSIONS: Conception scheduling or early pregnancy detection is required for better clinical counselling and the avoidance of unnecessary risk. PMID- 16268744 TI - [Antisense therapy in oncology: present situation]. AB - The purpose of antisense therapy is to control the regulation of genes contributing to cancer progression while sparing normal cell growth, which represents a novel alternative with fewer side effects when compared to conventional chemotherapy. Antisense oligonucleotides control cell proliferation by specifically blocking the expression of selected genes, and hence they are being developed as molecular drugs with potential activity for cancer treatment. Extensive clinical information and a number of clinical trials show encouraging results. This review discusses the most significant aspects of this new therapeutic alternative in oncology. Clinical trials performed thus far have demonstrated their short- to mid-term efficacy and safety; however, long-term studies are needed to definitely define their clinical effectiveness and true toxic profile. PMID- 16268746 TI - [Healthcare technologies assessment agencies. Utility of their assessment reports in a pharmacy department]. PMID- 16268745 TI - [Summary of therapy recommendations for fungal infection in the non-neutropenic critical-care patient: a multidisciplinary view]. PMID- 16268747 TI - [Likely serotoninergic syndrome from an interaction between amitryptiline, paroxetine, and linezolid]. PMID- 16268748 TI - [Fluphenazine decanoate overdose with no apparent adverse events]. PMID- 16268749 TI - [Acute coronary syndrome in a patient previously treated with capecitabine and 5 fluorouracil]. PMID- 16268750 TI - The effect of switching on compliance and persistence: the case of statin treatment. AB - OBJECTIVE: To determine the effect of switching drugs on the compliance and persistence of new statin users. STUDY DESIGN: Retrospective database analysis of pharmacy claims provided by a large pharmacy benefit manager. The study sample consisted of 38 866 new statin users 18 to 65 years old beginning treatment with atorvastatin calcium, fluvastatin sodium, lovastatin, pravastatin sodium, or simvastatin. METHODS: Compliance was measured by the "medication possession ratio," and persistence was measured by the time to discontinuation. Switching rates were derived from the proportions of patients filling a prescription other than the initial statin. RESULTS: Patients who switched statins were less compliant by 18.9% (odds ratio, 0.81; P < .001), as defined by the probability of having a medication possession ratio of 0.8 or higher, and were less persistent by 20.9% to 48.3% (P < .001) depending on the gap length used to define discontinuation. CONCLUSIONS: Switching statins substantially reduces the likelihood that patients will be compliant and remain on treatment long enough to obtain the full benefit of statin treatment. To ensure better compliance, special care should be given to patients who change drugs. PMID- 16268751 TI - Improving adherence to guidelines for hypertension drug prescribing: cluster randomized controlled trial of general versus patient-specific recommendations. AB - OBJECTIVE: To determine whether an intervention focusing clinician attention on drug choice for hypertension treatment improves concordance between drug regimens and guidelines. STUDY DESIGN: Cluster-randomized controlled trial comparing an individualized intervention with a general guideline implementation in geographically diverse primary care clinics of a university-affiliated Department of Veterans Affairs healthcare system. METHODS: Participants were 36 attending physicians and nurse practitioners (16 in the general group and 20 in the individualized group), with findings based on 4500 hypertensive patients. A general guideline implementation for all clinicians, including education about guideline-based drug recommendations and goals for adequacy of blood pressure control, was compared with addition of a printed individualized advisory sent to clinicians at each patient visit, indicating whether or not the patient's antihypertensive drug regimen was guideline concordant. We measured change from baseline to end point in the proportion of clinicians' patients whose drug therapy was guideline concordant. RESULTS: The individualized intervention resulted in an improvement in guideline concordance more than twice that observed for the general intervention (10.9% vs 3.8%, t = 2.796, P = .008). Bootstrap analysis showed that being in the individualized group increased the odds of concordance 1.5-fold (P = .025). The proportion of patients with adequate blood pressure control increased within each study group; however, the difference between groups was not significant. CONCLUSION: An individualized advisory regarding drug therapy for hypertension given to the clinician at each patient visit was more effective in changing clinician prescribing behavior than implementation of a general guideline. PMID- 16268752 TI - Continuity of care and cardiovascular risk factor management: does care by a single clinician add to informational continuity provided by electronic medical records? AB - BACKGROUND: Electronic medical records allow information sharing among multiple clinicians treating the same patient, enabling informational continuity between visits. OBJECTIVE: To assess the contribution of continuity of care (COC) with a single clinician to short-term outcomes in a setting in which electronic medical records are used. STUDY DESIGN: Retrospective cohort study. METHODS: Between January 1, 2003, and October 1, 2004, we identified 3718 patients assessed for lipid and blood pressure control and a subgroup of 1448 patients with diabetes mellitus assessed for glycemic control in the primary care clinics of a large Department of Veterans Affairs healthcare facility. Continuity of care was defined as having been seen by the same clinician (physician or nurse practitioner) in the year before testing. Analytic techniques accounting for clustering of patients by providers yielded robust estimators for the association between continuity with a single clinician and control of these cardiovascular disease risk factors. RESULTS: Patients with complete COC were more likely to be men with few medical problems and visits during the study period. Controlling for these differences, we detected no association between COC and patient attainment of recommended goals for cardiovascular disease risk factor control (P < .05 for all). CONCLUSION: Continuity of care with a single clinician contributes little to cardiovascular risk factor management in a setting in which electronic medical records provide enhanced informational continuity, although its value may be greater in the management and outcomes of established diseases that require coordination of care and ongoing collaboration between clinician and patient. PMID- 16268753 TI - Disease management produces limited quality-of-life improvements in patients with congestive heart failure: evidence from a randomized trial in community-dwelling patients. AB - BACKGROUND: Disease management programs are reported to improve clinical and quality-of-life outcomes while simultaneously lowering healthcare costs. OBJECTIVE: To examine the effectiveness of disease management in improving health related quality of life (HRQL) among patients with heart failure beyond 12 months. METHODS: A total of 1069 community-dwelling patients 18 years and older in South Texas with echocardiographic evidence of congestive heart failure were randomly assigned to disease management, augmented disease management, and control groups. They were followed up 18 months. Patients in the control group received usual care. Patients in the intervention groups were assigned a registered nurse as a disease manager who performed telephonic patient education and medication management. Health-related quality-of-life data (based on the Medical Outcomes Study 36-Item Short-Form Health Survey [SF-36]) were collected 4 times, at 6-month intervals. RESULTS: Disease management has a limited effect on HRQL. Analysis of the SF-36 health transition measure showed a positive effect of the intervention on self-reported improvement in health at 6 months and at 12 months (P = .04 and P = .004, respectively). However, no effect of disease management was observed across any of the SF-36 components. Women and patients with diastolic heart failure had poorer HRQL scores. CONCLUSIONS: Participation in disease management has little effect on HRQL outcomes in congestive heart failure. Beneficial effects on the SF-36 scale scores seen at 6 and 12 months were not sustained. Therefore, it is unclear whether disease management can provide long-term improvement in HRQL for patients with congestive heart failure. PMID- 16268754 TI - Tailored interventions to introduce influenza vaccination among 6- to 23-month old children at inner-city health centers. AB - BACKGROUND: Because of high rates of hospitalization for influenza infections among very young children (< 2 years), the Advisory Committee on Immunization Practices initiated a new policy in 2002 that encouraged vaccination of healthy children aged 6 to 23 months against influenza. OBJECTIVE: To evaluate the effectiveness of implementing tailored interventions to introduce influenza vaccination of children 6 to 23 months of age in inner-city practices. STUDY DESIGN: A before-after trial with historical and concurrent controls was conducted in 6 health centers in low-income urban locations. METHODS: Intervention sites were selected, and interventions (directed at 1534 patients who were 6 to 23 months old) were implemented from a menu of strategies. Vaccination rates were measured from medical record reviews. Focus groups of nursing staffs provided evaluative information on strategies. RESULTS: Influenza vaccination rates improved significantly at the intervention health centers compared with the control center. Preintervention (2001-2002) rates ranged from 0% to 7.6%, and intervention (2002-2003) rates ranged from 15.2% to 49.2% (P < .001). The number of interventions ranged from 6 to 11. Sites that used more interventions (odds ratio, 1.24; 95% confidence interval, 1.15-1.34) and had staff support of the vaccination effort (odds ratio, 1.91; 95% confidence interval, 1.40-2.60) had higher vaccination rates. CONCLUSIONS: Tailored interventions resulted in successful introduction of influenza vaccination of 6- to 23-month-old children in inner-city health centers. More strategies and enthusiastic staff support may result in higher vaccination rates. PMID- 16268755 TI - The effects of prescription drug cost sharing: a review of the evidence. AB - OBJECTIVES: To determine whether patients respond to increased cost sharing by substituting less expensive alternatives for medications with higher levels of copayments or coinsurance, and to examine the body of evidence on the relationships between cost sharing and use of essential or maintenance medications, health outcomes, process-of-care measures (such as medication adherence and discontinuation), and costs. STUDY DESIGN: Literature review. METHODS: Healthcare reference databases and key journals were searched to identify peer-reviewed empirical studies that examined the effects of variation in the amount of prescription drug copayments or coinsurance on healthcare utilization patterns. Thirty studies met our search criteria. RESULTS: Higher levels of prescription drug cost sharing generally produce intended effects, namely, decreasing the consumption of prescription drugs and steering patients away from nonpreferred to preferred brand-name drugs. However, patients do not always switch to generic drugs. Although not consistently reported, the most troublesome effects associated with higher levels of cost sharing are treatment disruptions (such as lower levels of treatment adherence, continuation, and initiation) for chronically ill patients. At times, higher levels of cost sharing can affect the use of essential medications and outcomes of care. CONCLUSIONS: Cost sharing reduces the consumption of prescription drugs but may have unintended effects on the process and outcomes of therapy. Further research is warranted in this area. The central question for health plan managers and policy makers is whether we will continue to use cost sharing as is or make modifications to reduce unintended effects. PMID- 16268756 TI - The dependence of the happiness of superstar scientists on the freedom to conduct animal experiments. PMID- 16268757 TI - QSAR applicabilty domain estimation by projection of the training set descriptor space: a review. AB - As the use of Quantitative Structure Activity Relationship (QSAR) models for chemical management increases, the reliability of the predictions from such models is a matter of growing concern. The OECD QSAR Validation Principles recommend that a model should be used within its applicability domain (AD). The Setubal Workshop report provided conceptual guidance on defining a (Q)SAR AD, but it is difficult to use directly. The practical application of the AD concept requires an operational definition that permits the design of an automatic (computerised), quantitative procedure to determine a models AD. An attempt is made to address this need, and methods and criteria for estimating AD through training set interpolation in descriptor space are reviewed. It is proposed that response space should be included in the training set representation. Thus, training set chemicals are points in n-dimensional descriptor space and m dimensional model response space. Four major approaches for estimating interpolation regions in a multivariate space are reviewed and compared: range, distance, geometrical, and probability density distribution. PMID- 16268758 TI - An approach to determining applicability domains for QSAR group contribution models: an analysis of SRC KOWWIN. AB - QSAR model predictions are most reliable if they come from the models applicability domain. The Setubal Workshop report provides a conceptual guidance for defining a (Q)SAR applicability domain. However, an operational definition is necessary for applying this guidance in practice. It should also permit the design of an automatic (computerised) procedure for determining a models applicability domain. This paper attempts to address this need for models that use a large number of descriptors (for example, group contribution-based models). The high dimensionality of these models imposes specific computational restrictions on estimating the interpolation region. The Syracuse Research Corporation KOWWIN model for prediction of the n-octanol/water partition coefficient is analysed as a case study. This is a linear regression model that uses 508 fragment counts and correction factors as descriptors, and is based on the group contribution approach. We conclude that the applicability domain estimation by descriptor ranges, combined with Principal Component rotation as a data pre-processing step, is an acceptable compromise between estimation accuracy and the amount of data in the training set. PMID- 16268759 TI - Psi-screen, an in vitro toxicity test system: applications in the bioassay of perfumes and fragrance chemicals. AB - The effects of 65 perfume formulations (perfume oils, perfumes, eau de parfum, eau de toilette) on mitochondrial membrane potential (Psim) and mitochondrial respiration have been investigated using a mitochondria-based assay for (Psim, termed Psi-Screen. All the perfume formulations tested are highly active in the Psi-Screen assay, and the major site of inhibition in all cases is NADH ubiquinone reductase (Complex I). This is confirmed in studies on the inhibition of NADH oxidase and NADH-ubiquinone reductase. Some formulations also inhibit succinate oxidation at either Complex II or Complex III. Evidence for the inhibition of mitochondrial ATPase is presented, as well as for the induction of reactive oxygen species production by perfume inhibition of Complex I. Thus, perfume formulations are multiple inhibitor mixtures which inhibit multiple bioenergetic functions at high dilutions. The implications of these findings are discussed with respect to cell toxicity via necrosis and/or apoptosis. Twenty candidate fragrance chemicals were investigated and all inhibited Complex I (5 at <35 microM). Mass screening strategies and high-throughput screening assays are discussed. PMID- 16268760 TI - The interpretation and application of the three Rs by animal ethics committee members. AB - The Three Rs form the basis of review of animal-use protocols by Animal Ethics Committees (AECs), but little research has examined how AECs actually interpret and implement the Three Rs. This topic was explored through in-depth, open-ended interviews with 28 members of AECs at four Canadian universities. In describing protocol review, AEC members rarely mentioned the Three Rs, but most reported applying some aspects of the basic concepts. Comments identified several factors that could impede full application of the Three Rs: incomplete understanding of the Three Rs (especially Refinement), trust that researchers implement Replacement and Reduction themselves, belief by some members that granting agency review covers the Three Rs, focus on sample size rather than experimental design to achieve Reduction, focus on harm caused by procedures to the exclusion of housing and husbandry, and lack of consensus on key issues, notably on the nature and moral significance of animal pain and suffering, and on whether AECs should minimise overall harm to animals. The study suggests ways to achieve more consistent application of the Three Rs, by providing AECs with up-to-date information on the Three Rs and with access to statistical expertise, by consensus-building on divisive issues, and by training on the scope and implementation of the Three Rs. PMID- 16268761 TI - Municipal sludge-induced phytotoxicity. AB - Municipal sludge (MS), containing various types of environmental pollutants, can exert phytotoxic effects in plants. Seed germination, chlorophyll content, plant weight, root and shoot growth, and metal levels, were measured as endpoints to assess the phytotoxic effects of leachates of MS from Lucknow city in three plants, Triticum aestivum, Brassica campestris and Phaseolus aureus. Among the five metals analysed, lead was found to be present at the highest concentration, and chromium at the lowest. Aqueous MS leachates had lower amounts of metals than pure MS. Seed germination was inhibited significantly in all three plants. The greatest inhibition of root and shoot growth was observed in B. campestris and T. aestivum. Total chlorophyll was depleted in all three plants, but the maximum depletion was observed in T.aestivum exposed to 10% leachate. Symptoms such as necrosis, leaf discoloration and root curling were also observed after exposure to 10% leachate, and plant weight was reduced significantly. It is evident that MS from Lucknow city exerted phytotoxic effects on all three test plant species. PMID- 16268762 TI - Comparison of an animal product free medium and normal growth supplement on the growth and barrier integrity of a human corneal epithelial cell line. AB - With the development of defined media for general and specific use with cell cultures, and concern over the use of human cells and over potential prion infections associated with growth factor extracts such as bovine pituitary extract, an animal product-free medium has become available. The basic keratinocyte defined medium can be used with a choice of animal product containing or animal product-free supplements. Human corneal epithelia cell lines were cultured in the media with these two types of supplement, and compared in terms of their growth rates, their capacity to form tight barriers, and calcium regulation of the location of a junction-associated protein, zonula occludins-1 (ZO-1). The growth rates were not different in the two media, as long as the recommended coating was applied to the culture flask for the animal product-free medium. The barrier function was equally effective for confluent cultures seeded at the same densities. A calcium concentration of 100 microM or above resulted in ZO-1 localisation at the cell membrane in either medium. Hence, cultures in the media are comparable, when the coating is employed. Further experiments are being conducted to establish the comparability of responses to chronic treatment with surfactants. PMID- 16268763 TI - The use of non-human primates in biological and medical research: evidence submitted by FRAME to the Academy of Medical Sciences/Medical Research Council/Royal Society/Wellcome Trust Working Group. AB - The Academy of Medical Sciences, the Medical Research Council, the Royal Society and the Wellcome Trust are undertaking a study into the use of non-human primates in biological and medical research. An independent working group of scientific experts, led by Sir David Weatherall, aims to produce a report summarising the findings of this study, early in 2006. The trends in primate research, and the nature and effects of recent and proposed changes in the global use of non-human primates in research, will be investigated. The associated ethical, welfare and regulatory issues, and the role and impact of the Three Rs principles of refinement, reduction and replacement will also be reviewed. As part of this study, a call for evidence was made. The evidence submitted by FRAME emphasised that the use of non-human primates for fundamental research or for regulatory testing still fails to take into account the fact that, although non-human primates are anatomically and physiologically similar to humans, they are not necessarily relevant models for studies on human disease or human physiology. FRAME continues to believe that we have a duty to ensure that these animals are not used without overwhelming evidence that they are the only suitable and relevant models for use in work of undeniable significance. PMID- 16268764 TI - The use of non-human primates in regulatory toxicology: comments submitted by FRAME to the Home Office. AB - The Home Office have circulated a document that summarises the discussions of a Primate Stakeholders Forum. The Forum took place in January 2004, and was convened to address the issues raised and the recommendations made in the Animal Procedures Committee 2002 report on the use of primates under the Animals (Scientific Procedures) Act 1986. The report emphasises the need for more resources focused on alternatives to toxicological testing in primates, including harmonising worldwide regulatory requirements, investigating the relevance of primate models, and improving the retrospective analysis of procedures involving primates. The document called for reasoned comments about the report to be submitted to the Home Office. In response, FRAME submitted a comprehensive paper, which evaluated each of the Animal Procedures Committees recommendations, along with the Home Office Forums comments. FRAME believes that, in coming to a decision as to whether primates should be used for regulatory testing, there must be full consideration of all the information available, including whether the ethological needs of any given species can be met prior to, during and following experimental use. Where these needs cannot be met, there must be a concerted effort to develop alternative models for research and testing. However, this should not detract from the ultimate goal of phasing out primate research altogether. PMID- 16268766 TI - The collaborative assessment and management of suicidality versus treatment as usual: a retrospective study with suicidal outpatients. AB - The Collaborative Assessment and Management of Suicidality (CAMS) is a novel clinical approach used to identify, assess, and manage suicidal outpatients (Jobes & Drozd, 2004). The results of a retrospective study evaluating the impact of CAMS versus treatment as usual (TAU) on suicidal outpatients are presented. Patients in the CAMS treatment group (n = 25) resolved their suicidality significantly more quickly than TAU patients (n = 30). CAMS was also significantly associated with decreased medical health care utilization in the 6 months after the start of suicide-related mental health treatment. These results provide promising preliminary support for the effectiveness of CAMS and a foundation for prospective research. PMID- 16268767 TI - Complicated grief and suicidal ideation in adult survivors of suicide. AB - While the prevalence of complicated grief has been demonstrated to be elevated in survivors of suicide, the association between complicated grief and suicidal ideation among adult survivors of suicide has not been explored. The purpose of the present study is to examine the association between complicated grief and suicidal ideation in suicide survivors. The Inventory of Complicated Grief and the Beck Depression Inventory were administered to 60 adult survivors within 1 month of a death by suicide of a family member or significant other. Complicated grief was associated with a 9.68 (CI: 1.036, 90.417) times greater likelihood of suicidal ideation after controlling for depression, suggesting that syndromal complicated grief heightens vulnerability to suicidal ideation. Clinicians may provide more comprehensive assessments by recognizing the possibility of suicidal ideation in those with complicated grief. PMID- 16268768 TI - Writing characteristics of suicidal people on the Internet: a psychological investigation of emerging social environments. AB - Three studies were carried out to investigate the writings of suicidal people on the Internet, hypothesizing consistent findings to equivalent research of offline writing. In Study 1, suicidal persons' (n = 34) attributions as the cause of their condition, as expressed in their free online writing, were compared to those of emotionally distressed, nonsuicidal (n = 16) and to nondistressed (n = 15) individuals. Results showed that suicidal persons have significantly more stable and global attributions than do the other groups. In Study 2, expressions of self-focus in online written messages were compared among suicidal, two levels of nonsuicidal but distressed, and nondistressed individuals (200 messages in each group). With the use of various expressions of self-focus, findings showed that suicidal people were distinctively self-focused in their writing, unlike their counterparts. In Study 3, psychologically relevant themes in the online writing of suicidal (n = 39) and highly distressed, nonsuicidal persons (n = 24) were compared. Results revealed that the former expressed significantly more unbearable psychological pain and cognitive constriction than did the latter. The findings of the three studies are consistent with those found in examinations of offline writing. The research has important implications for the use of online environments for psychological research and means for assessment, as well as for understanding suicidality. PMID- 16268769 TI - Suicidal and nonsuicidal adolescents: different factors contribute to self esteem. AB - Some risk and protective factors differ in their importance to suicidal and nonsuicidal people. In this research we explore the cross-sectional differences between risk factors among suicidal adolescents and nonsuicidal adolescents by focusing on self-esteem. Sixty-five suicidal and 390 nonsuicidal adolescents were compared on Harter's Self-Perceived Profile for Adolescents, self-concept stability, seeking support, loneliness, and depression. Self-concept stability, loneliness, and peer support correlated differently with self-esteem. In multivariate regression analyses, variance in self-esteem was explained by depression and loneliness, and among nonsuicidal adolescents also by self-concept stability, support, and competencies. Loneliness and self-concept stability related differently to self-esteem in suicidal and nonsuicidal adolescents. When the aim is to enhance self-esteem, this difference may delineate suicidal subgroups that need special interventions. PMID- 16268770 TI - Utah youth suicide study: psychological autopsy. AB - We conducted a psychological autopsy study to further understand youth suicide in Utah. While traditional psychological autopsy studies primarily focus on the administration of psychometric measures to identify any underlying diagnosis of mental illness for the suicide decedent, we focused our interviews to identify which contacts in the decedent's life recognized risk factors for suicidal behavior, symptoms of mental illness, as well as barriers to mental health treatment for the decedent. Parents and friends recognized most symptoms universally, although friends better recognized symptoms of substance abuse than any other contact. The study results suggest that parents and friends are the most appropriate individuals for gatekeeper training and, in conjunction with other innovative screening programs, may be an effective strategy in reducing adolescent suicide. PMID- 16268771 TI - Single and multiple suicide attempts and associated health risk factors in New Hampshire adolescents. AB - In this study we examined self-reported suicide attempts and their relationship to other health risk factors in a community sample of 16,644 adolescents. Fifteen percent endorsed suicide attempts (10% single; 5% multiple attempts) We hypothesized that multiple attempters would show higher prevalence of comorbid health risks than single or non-attempters. The three groups showed significant differences in ten health risk domains, on factors such as depressed mood, sexual assault, weight problems, and drug and alcohol use (ORs: 3.26-13.57). Repeated suicide attempts appear to be related to increased vulnerability and likelihood of harm in multiple domains of health risk. PMID- 16268772 TI - Factors associated with suicide attempts in female inmates: the hegemony of hopelessness. AB - In this study factors associated with past suicide attempts in female inmates were examined. Female inmate participants (N = 105) were given structured diagnostic assessments of antisocial and borderline personality disorders and substance dependence, as well as measures of depression, hopelessness, problem focused coping styles, and reasons for living. There was a high lifetime prevalence of past suicide attempts (38.1%). Suicide attempts were positively associated with personality disorders, hopelessness, depression, childhood physical/emotional abuse, and family history of suicide and mood disorders, and negatively associated with income, reasons for living, and problem-focused coping. Controlling for hopelessness, borderline personality disorder and family history of suicide attempts were the only variables that remained uniquely associated with suicide attempts. PMID- 16268773 TI - An evaluation of the Psychache Scale on an offender population. AB - This study examined the generalizability of a self-report measure of psychache to an offender population. The factor structure, construct validity, and criterion validity of the Psychache Scale was assessed on 136 male prison inmates. The results showed the Psychache Scale has a single underlying factor structure and to be strongly associated with measures of depression and hopelessness and moderately associated with psychiatric symptoms and the criterion variable of a history of prior suicide attempts. The variables of depression, hopelessness, and psychiatric symptoms all contributed unique variance to psychache. Discussion centers on psychache's theoretical application to the prediction of suicide. PMID- 16268774 TI - Self-mutilation and symptoms of depression, anxiety, and borderline personality disorder. AB - The goal of this study was to examine the relationship between self-mutilation and symptoms of depression and anxiety in a nonclinical population. Self mutilators reported significantly more symptoms of depression and anxiety than did the control group. When the group of self-mutilators was divided into individuals who cut themselves and individuals who harm themselves in other ways, we found that the between-group differences were primarily due to individuals with a history of cutting. Yet when symptoms of borderline personality disorder (BPD) were statistically controlled, all significant between-group differences in depressive and anxious symptoms were reduced to nonsignificant. These findings highlight the importance of assessing symptoms of BPD in self-mutilators, regardless of diagnosis. PMID- 16268775 TI - Living circumstances of suicide mortality in a South African city: an ecological study of differences across race groups and sexes. AB - In this study the importance of living area circumstances for suicide mortality was explored. Suicide mortality was assessed across race and sex groups in a South African city and the influence of area-based compositional and sociophysical characteristics on suicide risk was considered. Suicide mortality rates are highest among Whites, in particular White males. Main--and independent- dimensions of the living circumstances of residential areas (i.e., socioeconomic circumstances, economic need, and matrimony) influence age-adjusted suicide rates for both sexes but minimally so for race groups. Less favorable clusters of circumstances have a protective effect. PMID- 16268776 TI - Unacted suicide: a proposal. PMID- 16268777 TI - Recorded suicidality among patients with major depression in psychiatric outpatient care in Finland (1989-2001). PMID- 16268778 TI - Galphaq binds to p110alpha/p85alpha phosphoinositide 3-kinase and displaces Ras. AB - Several studies have reported that activation of G(q)-coupled receptors inhibits PI3K (phosphoinositide 3-kinase) signalling. In the present study, we used purified proteins to demonstrate that Galpha(q) directly inhibits p110alpha/p85alpha PI3K in a GTP-dependent manner. Activated Galpha(q) binds to the p110alpha/p85alpha PI3K with an apparent affinity that is seven times stronger than that for Galpha(q).GDP as measured by fluorescence spectroscopy. In contrast, Galpha(q) did not bind to the p110gamma PI3K. Fluorescence spectroscopy experiments also showed that Galpha(q) competes with Ras, a PI3K activator, for binding to p110alpha/p85alpha. Interestingly, co-precipitation studies using deletion mutants showed that Galpha(q) binds to the p85-binding domain of p110alpha and not to the Ras-binding domain. Expression of constitutively active Galpha(q)Q209L in cells inhibited Ras activation of the PI3K/Akt pathway but had no effect on Ras/Raf/MEK [MAPK (mitogen-activated protein kinase)/ERK (extracellular-signal-regulated kinase) kinase] signalling. These results suggest that activation of G(q)-coupled receptors leads to increased binding of Galpha(q).GTP to some isoforms of PI3K, which might explain why these receptors inhibit this signalling pathway in certain cell types. PMID- 16268780 TI - An analysis on gene architecture in human and mouse genomes. AB - A comparative genome analysis on exon-intron distribution profiles is performed for human and mouse genomes to deduce similarities and differences between them. Interestingly, both in human and mouse genomes, the total length in introns and intergenic DNA on each chromosome is significantly correlated to the chromosome size. The results presented provide a framework for understanding the nature and patterns of exon-intron length distributions, the constraints on them and their role in genome design and evolution. PMID- 16268781 TI - Natural antisense as potential regulator of alternative initiation, splicing and termination. AB - In humans an estimated 35-60% of genes are alternatively spliced. A large number of genes also show alternative initiation or termination. Regulation of these processes is still poorly understood. For alternative splicing it is believed that the relative concentration of certain proteins and the presence of certain regulatory elements are the key factors determining alterations in splicing pattern. However, there is evidence that antisense RNA might be part of the regulatory processes. Antisense RNA molecules could bind to the target pre-mRNA in a sequence-specific fashion, sterically blocking targeted splice sites and redirecting the spliceosome to available and unhindered splice sites. Here we describe an in silico investigation to identify human sense/antisense pairs with alternative initiation or termination in the sense gene and where only one of the isoforms overlaps the antisense transcript. Alternatively spliced genes with antisense transcripts covering the alternatively used splice site are also identified. Our analyses are based on the ASAP splicing annotation database from UCLA, the antisense transcripts data from Yelin et al., 2003, and the H invitational full-length cDNA database from JBIRC, Japan. These data gives new insight into the complexity of genomic organization and provide candidate loci for experimentalists to study antisense mediated regulation of alternative initiation, splicing and termination. Our result contains 468 clusters with this characteristic genomic organization and can be found at http://aistar.bii.a star.edu.sg/. PMID- 16268782 TI - A new domain family in the superfamily of alkaline phosphatases. AB - During the course of our large-scale genome analysis a conserved domain, currently detectable only in the genomes of Drosophila melanogaster, Caenorhabditis elegans and Anopheles gambiae, has been identified. The function of this domain is currently unknown and no function annotation is provided for this domain in the publicly available genomic, protein family and sequence databases. The search for the homologues of this domain in the non-redundant sequence database using PSI-BLAST, resulted in identification of distant relationship between this family and the alkaline phosphatase-like superfamily, which includes families of aryl sulfatase, N-acetylgalactosomine-4-sulfatase, alkaline phosphatase and 2,3-bisphosphoglycerate-independent phosphoglycerate mutase (iPGM). The fold recognition procedures showed that this new domain could adopt a similar 3-D fold as for this superfamily. Most of the phosphatases and sulfatases of this superfamily are characterized by functional residues Ser and Cys respectively in the topologically equivalent positions. This functionally important site aligns with Ser/Thr in the members of the new family. Additionally, set of residues responsible for a metal binding site in phosphatases and sulphtases are conserved in the new family. The in-depth analysis suggests that the new family could possess phosphatase activity. PMID- 16268783 TI - Analysis of gene ontology features in microarray data using the Proteome BioKnowledge Library. AB - Microarray technology has resulted in an explosion of complex, valuable data. Integrating data analysis tools with a comprehensive underlying database would allow efficient identification of common properties among differentially regulated genes. In this study we sought to compare the utility of various databases in microarray analysis. The Proteome BioKnowledge Library (BKL), a manually curated, proteome-wide compilation of the scientific literature, was used to generate a list of Gene Ontology (GO) Biological Process (BP) terms enriched among proteins involved in cardiovascular disease. Analysis of DNA microarray data generated in a study of rat vascular smooth muscle cell responses revealed significant enrichment in a number of GO BPs that were also enriched among cardiovascular disease-related proteins. Using annotation from LocusLink and chip annotation from the Gene Expression Omnibus yielded fewer enriched cardiovascular disease-associated GO BP terms. Data sets of orthologous genes from mouse and human were generated using the BKL Retriever. Analysis of these sets focusing on BKL Disease annotation, revealed a significant association of these genes with cardiovascular disease. These results and the extensive presence of experimental evidence for BKL GO and Disease features, underscore the benefits of using this database for microarray analysis. PMID- 16268779 TI - Induction of the permeability transition and cytochrome c release by 15-deoxy Delta12,14-prostaglandin J2 in mitochondria. AB - The electrophilic lipid 15-deoxy-Delta12,14-prostaglandin J2 (15d-PGJ2) is known to allow adaptation to oxidative stress in cells at low concentrations and apoptosis at high levels. The mechanisms leading to adaptation involve the covalent modification of regulatory proteins, such as Keap1, and augmentation of antioxidant defences in the cell. The targets leading to apoptosis are less well defined, but mitochondria have been indirectly implicated in the mechanisms of cell death mediated by electrophilic lipids. To determine the potential of electrophilic cyclopentenones to induce pro-apoptotic effects in the mitochondrion, we used isolated liver mitochondria and demonstrated that 15d-PGJ2 promotes Ca2+-induced mitochondrial swelling and cytochrome c release. The mechanisms involved are consistent with direct modification of protein thiols in the mitochondrion, rather than secondary formation of reactive oxygen species or lipid peroxidation. Using proteomic analysis in combination with biotinylated 15d PGJ2, we were able to identify 17 potential targets of the electrophile responsive proteome in isolated liver mitochondria. Taken together, these results suggest that electrophilic lipid oxidation products can target a sub-proteome in mitochondria, and this in turn results in the transduction of the electrophilic stimulus to the cell through cytochrome c release. PMID- 16268784 TI - MOLS--a program to explore the potential energy surface of a peptide and locate its low energy conformations. AB - We describe a computer program that uses mutually orthogonal Latin squares (MOLS) to perform an efficient and exhaustive conformational search of the multi dimensional potential energy hypersurface of an oligopeptide, and locate all its low energy conformations. The software package has been developed with a user friendly graphical interface using the Fast Light Tool Kit (FLTK)--a cross platform C++ toolkit. PMID- 16268785 TI - Folded RNA from an intron of one gene might inhibit expression of a counteracting gene. AB - Homeostatic maintenance of mRNA levels including prompt availability of mRNAs for translation in response to changing protein demands might be partly enabled by a system of combinatorial controls involving noncoding RNA blocking agents. This article proposes a specific version of that control mechanism, namely, a double stranded RNA folding from transcription of an intron of one gene might and leading to an agent that inhibits mRNA of a counteracting gene. Thus transcription of the first gene would automatically repress translation of the second. On the basis of a bioinformatics search, we suggest a possible example, namely, that pro-apoptosis gene PAR4 might inhibit anti-apoptosis gene XIAP. Part of an intron from PAR4 folds to form a large, stable hairpin, and reverse complement of the hairpin stem (with approximately 280 nucleotides) matches a sub sequence of an exon in XIAP. This would be part of an efficient system to drive initiated apoptosis to its conclusion. Figuratively speaking, it replaces two control knobs with one. Since repeats, some with many thousands of copies, occur throughout the genome with complex distributions, care must be taken before asserting that the presence of any repeat in any gene is significant. Our apoptosis example involves repeats, so experimental verification is needed and planned. However, if it is found that the noncoding RNA by-product of one gene folds into a hairpin that is processed into an agent that inhibits a counteracting gene, then the same type of control unit might be found extensively among counteracting families of genes of many types. PMID- 16268786 TI - CMDWave: conserved motifs detection using wavelets. AB - CMDWave (Conserved Motif Detection using WAVElets) is a web server that predicts conserved motifs in protein sequences. A set of query protein sequences are first aligned using ClustalW to obtain equal sized sequences. CMDWave then converts the sequences into a numerical representation using electron-ion interaction potential (EIIP). This is followed by a wavelet decomposition and reconstruction. A new similarity metric along with thresholding is then used to identify conserved motifs across all the query sequences. Users need not specify the number of motifs to be identified. For larger groups of sequences, results can be emailed to the users. PMID- 16268787 TI - KEGG-based pathway visualization tool for complex omics data. AB - Pathway-level visualization of omics data provides an essential means for systems biology, to capture the systematic properties of the inner activities of cells. Here we describe a web-based resource consisting of a web-application for the visualization of complex omics data onto KEGG pathways to overview all entities in the context of cellular pathways, and databases created with the software to visualize a series of microarray data. The web-application accepts transcriptome, proteome, metabolome, or the combination of these data as input, and because of this scalability it is advantageous for the visualization of cell simulation results. The web server can be accessed at http://www.g language.org/data/marray/. PMID- 16268788 TI - FASSM: enhanced function association in whole genome analysis using sequence and structural motifs. AB - We present an algorithm to detect remote homology, which arises through circular permutation and discontinuous domains. It is also helpful in detecting small domain proteins that are characterized by few conserved residues. The input to the algorithm is a set of multiply aligned protein sequence profiles. This method, coded as FASSM, examines the sequence conservation and positions of protein family signatures or motifs for the annotation of protein sequences and to facilitate the analysis of their domains. The overall coverage of FASSM is 93% in comparison to other validation tools like HMM and IMPALA. The method is especially useful for difficult relationships such as discontinuous domains during whole-genome surveys and is demonstrated to perform accurate family associations at sequence identities as low as 15%. PMID- 16268789 TI - An in silico mining for simple sequence repeats from expressed sequence tags of zebrafish, medaka, Fundulus, and Xiphophorus. AB - Teleost fish genome projects involving model species are resulting in a rapid accumulation of genomic and expressed DNA sequences in public databases. The expressed sequence tags (ESTs) collected in the databases can be mined for the analysis of both structural and functional genomics. In this study, we in silico analyzed 49,430 unigenes representing a total of 692,654 ESTs from four model fish for their potential use in developing simple sequence repeats (SSRs), or microsatellites. After bioinformatical mining, a total of 3,018 EST derived SSRs (EST-SSRs) were identified for 2,335 SSR containing ESTs (SSR-ESTs). The frequency of identified SSR-ESTs ranged from 1.5% for Xiphophorus to 7.3% for zebrafish. The dinucleotide repeat motif is the most abundant SSR, accounting for 47%, 52%, 64%, and 78% for medaka, Fundulus, zebrafish, and Xiphophorus, respectively. Simulation analysis suggests that a majority of these EST-SSRs have sufficient flanking sequences for polymerase chain reaction (PCR) primer design. Comparative DNA sequence analyses of SSR-ESTs identified several cross-species SSRs and sequences that may be used as cross-reference genes in comparative studies. For example, the flanking sequences of one SSR (CTG)n within the pituitary tumor-transforming gene (PTTG) 1 interacting protein (PTTGIP), showed conservation spanning the medaka, Fundulus, human, and mouse genomes. This study provides a large body of information on EST-SSRs that can be useful for the development of polymorphic markers, gene mapping, and comparative genome analysis. Functional analysis of these SSR-ESTs may reveal their role in metabolism and gene evolution of these model species. PMID- 16268790 TI - Correlation Finder. AB - Correlation Finder is a free software which allows to seek exhaustively correlations between nucleotides in genomic sequences. It permits to analyze generic DNA sequences and genic sequences where the codon phase needs to be taken into account. Its graphic interface allows to easily set the parameters that characterize the motifs being sought. This tool handles large data sets and runs on the Windows operative system. PMID- 16268791 TI - Accessing optimal primer distance from insert. AB - When building either DNA or cDNA libraries, a researcher looks at the vector multiple cloning site and chooses which restriction enzyme(s) will be used to clone the inserts. Although this procedure does not seem to be important, the accurate choosing of primer to insert distance can save time and money from genome and transcriptome projects. Here, 846 single-pool pUC18 sequences were produced and compared with the pUC18 consensus using local alignment tools. Data show that reads often contain 0-20 miscalled bases at the beginning of read and noise to signal transition is frequently found at 46-54 bases from the first 3' base downstream the sequencing primer. For SWAT-based approaches, 60 bases was the distance where over 90% of the sequences provided reliable information, presenting 13 vector bases on average. Looking at the data, it is possible to choose the most appropriate primer to insert distance for many applications. PMID- 16268792 TI - Computing TaqMan probes for multiplex PCR detection of E. coli O157 serotypes in water. AB - Diarrheagenic E. coli strains contribute to water related diseases in urban and rural environment in developing and developed world. E. coli pathotype and pathogenicity varies due to complex multifactorial mechanism involving a large number of virulence factors. Rapid assessment of the virulence pattern of E. coli isolates is possible by Real-Time PCR probes like TaqMan. For designing TaqMan probes and primers for multiplex PCR selected E. coli gene sequences: stx1, stx2, hlyA, chuA, eae, lacZ, lamB and fimA were retrieved from NCBI's GenBank database. The alignment of the multiple sequences and analysis of conserved sequences was carried out using ClustalW and BLAST programs. The primers and Taqmen probes were designed using Beacon Designer software version 2.1 for two multiplexed PCR assays. In silico PCR simulation of these assays showed PCR products for stx2 (248bp) stx1 (102 bp), lacZ (228bp) and lamB (86 bp) in multiplex #1 and eae (200bp), chuA (147 bp), hlyA (141bp) and fimA (79 bp) in multiplex #2, respectively. These multiplexed PCR amplification products and probes can be used to identify and confirm presence of O157:H7/ H7-, O157:H43/45 and O26:H-/H11 serotypes. In conclusion, multiplex Real-Time Polymerase Chain Reaction oligomers and TaqMan probes designed and validated in silico will be helpful in management of water quality and outbreaks, by improving specificity and minimizing time needed for in vitro verification work. PMID- 16268793 TI - T cell receptor/peptide/MHC molecular characterization and standardized pMHC contact sites in IMGT/3Dstructure-DB. AB - One of the key elements in the adaptive immune response is the presentation of peptides by the major histocompatibility complex (MHC) to the T cell receptors (TR) at the surface of T cells. The characterization of the TR/peptide/MHC trimolecular complexes (TR/pMHC) is crucial to the fields of immunology, vaccination and immunotherapy. In order to facilitate data comparison and cross referencing between experiments from different laboratories whatever the receptor, the chain type, the domain, or the species, IMGT, the international ImMunoGeneTics information system (http://imgt.cines.fr), has developed IMGT ONTOLOGY, the first ontology in immunogenetics and immunoinformatics. In IMGT/3Dstructure-DB, the IMGT three-dimensional structure database, TR/pMHC molecular characterization and pMHC contact analysis are made according to the IMGT Scientific chart rules, based on the IMGT-ONTOLOGY concepts. IMGT/3Dstructure-DB provides the standardized IMGT gene and allele names (CLASSIFICATION), the standardized IMGT labels (DESCRIPTION) and the IMGT unique numbering (NUMEROTATION). As the IMGT structural unit is the domain, amino acids at conserved positions always have the same number in the IMGT databases, tools and Web resources. For the TR alpha and beta chains, the amino acids in contact with the peptide/MHC (pMHC) are defined according to the IMGT unique numbering for V-DOMAIN. The MHC cleft that binds the peptide is formed by two groove domains (G-DOMAIN), each one comprising four antiparallel beta strands and one alpha helix. The IMGT unique numbering for G-DOMAIN applies both to the first two domains (G-ALPHA1 and G-ALPHA2) of the MHC class I alpha chain, and to the first domain (G-ALPHA and G-BETA) of the two MHC class II chains, alpha and beta. Based on the IMGT unique numbering, we defined eleven contact sites for the analysis of the pMHC contacts. The TR/pMHC contact description, based on the IMGT numbering, can be queried in the IMGT/StucturalQuery tool, at http://imgt.cines.fr. PMID- 16268794 TI - Ontology based standardization of Petri net modeling for signaling pathways. AB - Taking account of the great availability of Petri nets in modeling and analyzing large complicated signaling networks, semantics of Petri nets is in need of systematization for the purpose of consistency and reusability of the models. This paper reports on standardization of units of Petri nets on the basis of an ontology that gives an intrinsic definition to the process of signaling in signaling pathways. PMID- 16268795 TI - Molecular analysis of a 348 base-pair segment of open reading frame 2 of human astrovirus. A characterization of Colombian isolates. AB - We are reporting computational studies of several genotyped strains of astrovirus isolated in Colombia which we have genotyped using the 348-bp segment located between nucleotides 258 and 606 close to the amino terminal region of the complete ORF 2. By biocomputational techniques this 348-bp segment from the different strains was translated into an amino acid sequence. The sequences were aligned and compared in order to build a dendrogram. Our results show that the 348 bp and the 116 amino acid peptides cluster in a very conservative way, showing slight genetic variations between them. This slight sequence variability does not allow us to identify common amino acid substitution patterns shared by all members of each HAstV specific type, thus suggesting that antigenic epitopes are probably located outside the 116 peptide fragment of this capsid protein. These results show that there is little recombination among different regions of the ORF2, thus suggesting that this genotyping method should continue to be useful for serotyping HAstV isolates. PMID- 16268796 TI - Signal-theoretical DNA similarity measure revealing unexpected similarities of E. coli promoters. AB - We present an implementation of the signal theory based approach for detection of novel types of DNA similarity which are based on physical properties of DNA. Systematic study of the sensitivity of the new similarity measure revealed qualitative differences to letter-based similarity. A variety of physical parameters of DNA double strands, which in a straightforward way reflect different kinds of information hidden behind the primary structure of DNA, showed a wide range of recognition power of the signal similarity measure. We applied the novel DNA similarity measure for the analysis of promoters of E.coli genes. We found that promoter similarities revealed by our approach correlate with their transcription regulatory responsivenesses to different antibiotic and osmotic treatments. Accelerated by special hardware for fast Fourier transformations, the method is easily applicable for the analysis of entire eukaryotic genomes in minutes. PMID- 16268797 TI - I-superfamily conotoxins: sequence and structure analysis. AB - I-superfamily conotoxins have four-disulfide bonds with cysteine arrangement C-C CC-CC-C-C, and they inhibit or modify ion channels of nerve cells. They have been characterized only recently and are relatively less well studied compared to other superfamily conotoxins. We have detected selective and sensitive sequence pattern for I-superfamily conotoxins. The availability of sequence pattern should be useful in protein family classification and functional annotation. We have built by homology modeling, a theoretical structural 3D model of ViTx from Conus virgo, a typical member of I-superfamily conotoxins. The modeling was based on the available 3D structure of Janus-atracotoxin-Hv1c of Janus-atracotoxin family whose members have been suggested as possible biopesticides. A study comparing the theoretically modeled structure of ViTx, with experimentally determined structures of other toxins, which share functional similarity with ViTx, reveals the crucial role of C-terminal region of ViTx in blocking therapeutically important voltage-gated potassium channels. PMID- 16268798 TI - Why is the management of glucocorticoid deficiency still controversial: a review of the literature. AB - All endocrinologists would like to make glucocorticoid replacement therapy for their hypoadrenal patients as physiological as possible. Many would like the reassurance of a method of monitoring such treatment to confirm that they are achieving this aim. Advances in our knowledge of the normal physiology are relevant to our attempts to do this. The cortisol production rate in normal subjects is lower than was previously believed. The normal pattern of glucocorticoid secretion includes both a diurnal rhythm and a pulsatile ultradian rhythm. Glucocorticoid access to nuclear receptors is 'gated' by the 11-beta hydroxysteroid dehydrogenase enzymes, which interconvert active cortisol and inactive cortisone. Such complexities make the target of physiological glucocorticoid replacement therapy hard to achieve. The available evidence suggests that conventional treatment of hypoadrenal patients may result in adverse effects on some surrogate markers of disease risk, such as a lower bone mineral density than age-sex matched controls, and increases in postprandial glucose and insulin concentrations. Although the quality of life of hypoadrenal patients may be impaired, there is no evidence of an improvement on higher doses of steroids, although quality of life is better if the hydrocortisone dose is split up, with the highest dose taken in the morning. Thus the evidence suggests that most patients may safely be treated with a low dose of glucocorticoid (e.g. 15 mg hydrocortisone daily) in two or three divided doses, with education about the appropriate action to take in the event of intercurrent illnesses. PMID- 16268799 TI - Increased plasma N-terminal pro-B-type natriuretic peptide and markers of inflammation related to atherosclerosis in patients with primary hyperparathyroidism. AB - OBJECTIVE: Increased risk of cardiovascular disease has been reported in patients with primary hyperparathyroidism (PHPT). The aim of this study was to evaluate novel plasma risk markers of cardiovascular disease in patients with PHPT. DESIGN: PHPT patients were evaluated with a control group. Patients who underwent parathyroidectomy were re-evaluated after 7 and 18 months. PATIENTS: Forty-five PHPT patients and 40 matched controls participated. Seventeen patients underwent parathyroidectomy. MEASUREMENTS: Plasma N-terminal pro-B-type natriuretic peptide (NT-proBNP), high-sensitivity C-reactive protein (CRP), interleukin-6 (IL-6) and tumour necrosis factor alpha (TNF-alpha), lipids and blood pressure were measured. In 27 patients a bicycle exercise test and radionuclide angiography were performed, and repeated in those who underwent parathyroidectomy. RESULTS: Plasma NT-proBNP, CRP and TNF-alpha, but not IL-6, were higher in patients with PHPT than in controls (P < 0.01 and P = 0.17, respectively). In patients with PHPT, NT-proBNP correlated with systolic blood pressure, left ventricular end diastolic volume, and peak oxygen uptake (all P < 0.01). Log CRP correlated with systolic and diastolic blood pressure (both P < 0.05) and log IL-6 (P < 0.01). No significant correlations were observed between PTH or calcium and risk markers of cardiovascular disease. No decrease in NT-proBNP, markers of inflammation or blood pressure was observed after parathyroidectomy. CONCLUSIONS: Our data suggest that hypertension or other factors, rather than plasma calcium or PTH, could explain the increased levels of the inflammatory markers and NT-proBNP in PHPT. We therefore suggest that aggressive treatment of hypertension should be initiated in patients with PHPT to try to reduce the increased cardiovascular mortality described in PHPT. Further prospective studies are needed to validate the suggestion that increased levels of NT-proBNP and inflammatory markers also represent strong prognostic markers of cardiovascular disease in patients with PHPT. PMID- 16268800 TI - Adrenocortical insufficiency after pituitary surgery: an audit of the reliability of the conventional short synacthen test. AB - BACKGROUND: Assessment of the hypothalamic-pituitary-adrenal (HPA) axis after pituitary surgery is important for appropriate decision making regarding replacement therapy. The synacthen test is often used but is questioned, as time has to elapse for adrenal atrophy to develop. OBJECTIVE: To audit the use of the 250 microg synacthen test after transsphenoidal adenomectomy. METHODS: A retrospective study of 110 patients submitted to first-time transsphenoidal adenomectomy. Anterior pituitary testing was performed preoperatively, 1 week and 1, 3, 6 and 12 months postoperatively. The adrenocortical function was tested by a synacthen test (250 microg synacthen i.v.). RESULTS: Thirty-two out of 71 patients with normal HPA function before surgery developed insufficiency postoperatively, seven patients presenting an insufficient test response after 1 week, 16 after 1 month and nine after 3 months, whereas none became insufficient during the remaining follow-up. Three patients presented symptomatic adrenal insufficiency during the first postoperative week despite a normal test. All of these developed an insufficient test 1 month postoperatively. A 1-week basal plasma cortisol > 400 nmol/l indicated HPA sufficiency, whereas a basal cortisol < or = 100 nmol/l indicated insufficiency when related to the diagnosis based on the 3-month synacthen test. CONCLUSION: This study confirms that the synacthen test is of limited use in the early postoperative phase, because out of 62 patients with normal 1-week postoperative synacthen responses, 23 patients developed a test that was indicative of adrenal insufficiency over 1-3 months. Our results indicate that a large proportion of patients should be considered for hydrocortisone replacement therapy up to 3 months postoperatively followed by reassessment of the HPA axis. PMID- 16268801 TI - Vitamin D status, seasonal variations, parathyroid adenoma weight and bone mineral density in primary hyperparathyroidism. AB - BACKGROUND: Primary hyperparathyroidism (PHPT) and vitamin D insufficiency are common conditions that can occur in combination. However, low plasma 25 hydroxyvitamin D (25OHD) may also enhance the risk of PHPT or modify disease severity. AIM: To compare the risk of vitamin D insufficiency and deficiency stratified by age, sex and season between PHPT patients and controls and to assess associations between plasma 25OHD and adenoma weight, biochemical variables, bone mineral density (BMD) and clinical complications. DESIGN: Cross sectional study. MATERIAL: A total of 289 consecutive Caucasian patients with PHPT aged 65.9 (24-92) years, 289 sex-, age- and season-matched normocalcaemic controls and 187 healthy adult blood donors. PHPT diagnosis was confirmed in 214 by neck exploration. RESULTS: Vitamin D insufficiency (plasma 25OHD < 50 nmol/l) was observed in 81% of PHPT patients compared with 60% of sex- and age-matched controls (P < 0.001) and 35% of blood donors (P < 0.001). During summer, 77%vs. 53% (P < 0.001) and 4% (P < 0.001), respectively, had vitamin D insufficiency. Average plasma 25OHD was 41 (range 9-87) nmol/l among 27 PHPT patients compared with 87 (21-173) nmol/l (P < 0.001) among aged-matched blood donors. During winter, 86%vs. 66% (P < 0.001) and 71% (P < 0.05), respectively, had vitamin D insufficiency. Vitamin D deficiency (plasma 25OHD < 25 nmol/l) was observed in 33% of PHPT patients compared with 20% of age- and sex-matched controls (P < 0.001) and 13% of blood donors (P < 0.001). Both PHPT patients and controls showed seasonal variations in 25OHD related to the average number of sun hours, but values were lower in PHPT patients at all calendar months. In PHPT patients low plasma 25OHD was associated with higher plasma levels of calcium, PTH and alkaline phosphatase and with lower renal calcium excretion, femoral neck and forearm BMD. No association was found between plasma 25OHD and adenoma weight (total or divided into tertiles). There was a trend towards increased risk of osteoporotic fractures (P < 0.08) with low plasma 25OHD. CONCLUSION: Vitamin D insufficiency and deficiency are common findings in PHPT and occur more often than in a sex- and age-matched control group referred from general practice and in normal blood donors irrespective of season. Low plasma 25OHD levels are associated with an aggravated clinical presentation of PHPT but do not affect adenoma size. PMID- 16268802 TI - Pharmacokinetic profile of lanreotide Autogel in patients with acromegaly after four deep subcutaneous injections of 60, 90 or 120 mg every 28 days. AB - OBJECTIVE: To investigate the pharmacokinetic profile of a prolonged release, aqueous Autogel formulation of the somatostatin analogue lanreotide (Lan-ATG). DESIGN: A phase II, randomized, double-blind study, during which patients received 60, 90 or 120 mg Lan-ATG for four fixed administrations at 28-day intervals. PATIENTS: A total of 18 patients with acromegaly were recruited; six patients were randomized to each treatment. MEASUREMENTS: Lanreotide minimum concentration (C(min)), maximum serum concentration (C(max)) and area under the concentration-time curve during a dosing interval (AUC(tau)) were assessed after a single dose and at steady state (ss). Serum GH and IGF-1 levels were assessed before each administration and at the end of the study. RESULTS: After a single administration, dose proportionality for C(min,1), C(max) and AUC(tau) was demonstrated statistically. After repeated administrations, Lan-ATG exhibited linear pharmacokinetics over the dose range and ss values of C(min), C(max) and AUC(tau) increased in a dose-dependent, linear manner. Mean C(max,ss) values were only two- to fourfold greater than C(min,ss) values, and there was good control over the entire release profile. Serum levels of GH and IGF-1 declined over the course of the study and acromegaly symptoms improved. The treatment was well tolerated. CONCLUSIONS: Lan-ATG showed linear pharmacokinetic profiles over the three dose levels after both single and repeated dosing, no initial burst effect and good control over the entire release profile. Despite the absence of dose adaptation, four injections of Lan-ATG were effective in lowering serum levels of GH and IGF-1. PMID- 16268803 TI - Macroprolactin does not contribute to elevated levels of prolactin in patients on renal replacement therapy. AB - OBJECTIVE: Three molecular forms of PRL with molecular weights of 23, 50-60 and > 100 kDa have been defined. The high-molecular-weight forms are called macroprolactin. Different immunoassays produce varyingly elevated results with macroprolactin-containing sera. The kidneys are reported to clear 25% of PRL from the circulation. Hyperprolactinaemia is seen in 20-75% of patients with chronic renal failure (CRF). PRL clearance rate has been reported to be reduced in CRF and the resulting hyperprolactinaemia is due to reduced renal function. PATIENTS: To determine the contribution of macroprolactinaemia to elevated PRL levels in CRF, 91 patients receiving haemodialysis (HD), continuous ambulatory peritoneal dialysis (CAPD) and renal transplantation (RT) therapies and 72 control subjects were included in the study. MEASUREMENTS: Serum PRL levels were measured by a sandwich immunoassay with electrochemical detection. Following polyethylene glycol (PEG) precipitation, recovery ratios were calculated and samples with a recovery of < 50% were identified as having macroprolactin isoforms. RESULTS: The serum and supernatant PRL levels of CRF patients were significantly higher than those of the control group (P < 0.001). The serum PRL levels of HD and CAPD patients were significantly higher than those of the RT patients (P < 0.001). The serum PRL levels of the RT patients and the control group did not differ significantly (P > 0.05). A moderate correlation was found between PRL and creatinine levels (r = 0.609, P < 0.001). CONCLUSIONS: The hyperprolactinaemia seen in renal replacement therapy is not associated with the presence of macroprolactin isoforms but with the decline in renal function. PMID- 16268804 TI - Diabetic neuropathy is associated with activation of the TNF-alpha system in subjects with type 1 diabetes mellitus. AB - OBJECTIVE: The development of diabetic neuropathy (DN) is predicted by cardiovascular risk factors associated with insulin resistance. As inflammation seems to be implicated in the pathogenesis of insulin resistance, we investigated whether subjects with type 1 diabetes mellitus (T1DM) and DN have an increase in plasma concentrations of inflammatory proteins involved in insulin resistance. DESIGN: Cross-sectional. Patients One hundred twenty subjects, all diagnosed with T1DM 14 years before. MEASUREMENTS: (1) Sex, age, body mass index, waist-to-hip ratio (WHR), blood pressure, smoking, alcohol intake, insulin dose, HbA1c and lipid profile; (2) DN (peripheral and cardiac autonomic), retinopathy and nephropathy; (3) plasma concentrations of soluble fractions of tumour necrosis factor alpha receptors 1 and 2 (sTNFR1 and sTNFR2), interleukin-6, high-sensitive C-reactive protein, adiponectin and leptin; and (4) insulin resistance (by way of a mathematical estimation of the glucose disposal rate - eGDR-). RESULTS: Thirty six subjects had DN and 84 did not. Subjects with DN received higher insulin doses (57.6 +/- 16.7 vs. 49.2 +/- 15.0 IU/day; P = 0.008) and had higher WHR (0.85 +/- 0.07 vs. 0.81 +/- 0.10; P = 0.007) and HbA1c values (8.5 (7.6-9.6) vs. 7.7 (7.3-8.9)%; P = 0.049) than subjects without DN. They also had higher values of sTNFR1 (2.42 +/- 0.60 vs. 1.96 +/- 0.66 microg/l; P = 0.001) and sTNFR2 (4.73 +/- 1.33 vs. 4.14 +/- 1.09 microg/l; P = 0.015), and were more insulin resistant (eGDR values: 7.28 (5.83-8.03) vs. 8.30 (7.17-9.03) mg kg(-1) min(-1); P = 0.003). The relationship between DN and either sTNFR1 or sTNFR2 remained essentially unchanged after adjusting for several confounders, including glycaemic control, WHR, lipid profile, blood pressure and other microvascular complications (OR for sTNFR1: 2.592 (1.222-5.498), P = 0.013; OR for sTNFR2: 2.124 (1.258-3.587), P = 0.005). CONCLUSIONS: The activity of the TNF-alpha system is increased in subjects with type 1 diabetes mellitus and diabetic neuropathy, regardless of their glycaemic control and cardiovascular risk factors associated with insulin resistance. These results suggest that TNF-alpha may play a pathogenic role in the development of diabetic neuropathy. PMID- 16268805 TI - Adaptation of the hypothalamic-pituitary hormones during intensive endurance training. AB - OBJECTIVE: Physical activity leads to changes in the hypothalamic-pituitary hormonal system. However, acute and long-term adaptations have not yet been precisely characterized. In this study, the changes of the hormonal system as a result of marathon training and running a marathon were examined. In particular, we focused on adaptations of the hypothalamic-pituitary-adrenocortical (HPA) axis, regarding the activation or inactivation of cortisol to cortisone by the 11beta-hydroxysteroid-dehydrogenase system (11beta-HSD). DESIGN: Patient measurements: 8 healthy women and 11 healthy men volunteered for this study. Blood samples, 24-h urine and a dexamethasone suppression test were analysed for metabolic and hormonal parameters at five different dates 12 weeks around a marathon. RESULTS: Cortisol and ACTH values decreased significantly 2 days after the marathon, whereas the activity of the whole body 11beta-HSD-1 was up regulated. An increased suppression of cortisol levels was observed in the dexamethasone suppression test after 6 weeks of reduced training levels. Ghrelin was elevated 2 days after the marathon. Only minor changes in the other hypothalamic-pituitary-hormonal axes could be observed. However, the free androgen index increased significantly after 6 weeks of reduced training. CONCLUSIONS: The HPA system appeared to become chronically activated by continuous physical training and therefore less sensitive to the dexamethasone suppression test. The acute stress of the marathon led to a central exhaustion of the HPA system with a paracrine counteraction by the activation of the 11beta-HSD system. Changes in the other hypothalamic-pituitary hormonal axes were the result of long-term differences in training levels and were not altered by the marathon. PMID- 16268806 TI - Alterations in the oxidant-antioxidant status in prepubertal children with growth hormone deficiency: effect of growth hormone replacement therapy. AB - OBJECTIVE: Growth hormone deficiency (GHD) in adults is associated with increased oxidative stress determined by the underlying GH-IGF-1 axis alterations. Despite GHD being a common diagnosis in children with short stature, no data on the oxidant/antioxidant status are available in this age group. This study was designed to detect differences in oxidative stress parameters between prepubertal GH-deficient children and healthy controls. Furthermore, the effect of 12 months of conventional GH replacement (rGH) on oxidant-antioxidant status was evaluated in the GHD group only. PATIENTS: Ten (nine males and one female) prepubertal children (mean age 9.1 +/- 1.3 years) with GHD were recruited and matched for sex and age (9.2 +/- 1.9 years) with 20 healthy controls (18 males and two females). MEASUREMENTS: At study entry, lag phase, an index of susceptibility of low density lipoprotein (LDL) to in vitro oxidation, malondialdehyde (MDA) and vitamin E were measured in all subjects. These parameters were also evaluated in GH-deficient children after 12 months of rGH treatment. RESULTS: The lag phase was significantly decreased in GH-deficient children compared to healthy controls (15.50 +/- 7.4 vs. 43.00 +/- 9.2 min; P = 0.0007), while MDA was significantly increased (1.33 +/- 0.38 vs. 0.46 +/- 0.10 nmol/mg; P = 0.0006). Vitamin E levels were significantly decreased (22.44 +/- 9.57 vs. 35.38 +/- 16.49 micromol/l; P = 0.001). IGF-1 and IGFBP-3 correlated directly to lag phase (r = 0.48; P = 0.01; r = 0.63, P = 0.002, respectively) and to vitamin E (r = 0.59, P = 0.003; r = 0.58, P = 0.006, respectively). By contrast, IGF-1 and IGFBP-3 correlated indirectly to MDA (r = -0.47, P = 0.01; r =-0.65, P = 0.002, respectively). After 1 year of rGH therapy, lag phase (39.32 +/- 15.24 min; P = 0.005) and vitamin E (34.9 +/- 7.7 micromol/l; P = 0.005) increased significantly, while MDA decreased significantly (0.71 +/- 0.42 nmol/mg; P = 0.005), reaching normal levels. CONCLUSIONS: These data show that children with GHD have substantially increased oxidative stress parameters compared to healthy controls and demonstrate a normalization of these parameters after 1 year of rGH therapy. PMID- 16268807 TI - Determination of iodine concentration in urine by isotope dilution analysis and thyroid volume of school children in the west coast of Turkey after mandatory salt iodization. AB - OBJECTIVE: This study was designed to evaluate iodine deficiency status in children 6-12 years in the west coast (Aegean Region) of Turkey after 5 years of mandatory iodine prophylaxis. A total of 2300 children from 72 populations (rural and urban area) were evaluated with urinary iodine excretion and thyroid volume. DESIGN: Cross-sectional, observational study designed and performed according to surveillance methods for iodine deficiency disease (IDD) prevalence recommended by WHO/UNICEF/ICCIDD. SUBJECTS: The study population consisted of 2300 school children age ranging from 6 to 12 years. The children were selected by multiple stage randomization from 91 primary schools of 76 zones (91 clusters). Information about the use of iodized salt was obtained from the families. MEASUREMENTS: Data on the following were collected: birth date, sex, weight, height, thyroid size by palpation and ultrasonography; and urinary iodine by isotope dilution analysis method. Thyroid volumes above 97th percentile according to the WHO/ICCIDD by age and body surface area (BSA) were accepted as goitre. RESULTS: Iodized salt consumption was 51.7%. The prevalence of goitre determined by palpation was 12.1% and by ultrasound based on BSA and age were 9.8% and 5.5%, respectively. Median urinary iodine was 53 (2-142) microg/l. CONCLUSION: Mild to severe degree of iodine deficiency was detected in the west coast of Turkey. PMID- 16268808 TI - Long-term remission rates after pituitary surgery for Cushing's disease: the need for long-term surveillance. AB - OBJECTIVE: There have been a few reports on long-term remission rates after apparent early remission following pituitary surgery in the management of Cushing's disease. An undetectable postoperative serum cortisol has been regarded as the result most likely to predict long-term remission. Our objective was to assess the relapse rates in patients who underwent transsphenoidal surgery in order to determine whether undetectable cortisol following surgery was predictive of long-term remission and whether it was possible to have long-term remission when early morning cortisol was measurable but not grossly elevated. Endocrinological factors associated with late relapse were also studied. PATIENTS: We reviewed the long-term outcome in 63 patients who had pituitary surgery for the treatment of Cushing's disease between 1979 and 2000. MEASUREMENTS: Case notes were reviewed and the current clinical and biochemical status assessed. Our usual practice was that early after the operation, an 08:00 h serum cortisol was measured 24 h after the last dose of hydrocortisone. This was followed by a formal low-dose dexamethasone suppression test. Current clinical status and recent 24-h urinary free cortisol values were used as an index of activity of the Cushing's disease. If there was evidence suggesting relapse, a low-dose dexamethasone suppression test was performed. In many patients, sequential collections of early morning urine specimens for urinary cortisol to creatinine ratio were also performed in an attempt to diagnose cyclical and intermittent forms of recurrent hypercortisolism. We did this if there was conflicting endocrine data, or if patients were slow to lose abnormal clinical features. RESULTS: Mean age at diagnosis was 40.3 years (range 14-70 years). Mean follow-up up time was 9.6 years (range 1-21 years). Forty-five patients (9 males/36 females) achieved apparent remission immediately after surgery and were subsequently studied long term. Of these 45 patients, four have subsequently died while in remission from hypercortisolism. Ten of the remaining 41 patients have relapsed. Of those 10, six demonstrated definite cyclical cortisol secretion. Two of the 10 had undetectable basal serum cortisol levels in the immediate postoperative period. Thirty-one patients are still alive and in remission. Fourteen (45%) of the 31 who remained in remission had detectable serum cortisol levels (> 50 nmol/l) immediately postoperatively, and remain in remission after a mean of 8.8 years. Our relapse rate was therefore 10/45 (22%), after a mean follow-up time of 9.6 years, with mean time to relapse 5.3 years. CONCLUSIONS: The overall remission rate of 56% (35/63) at 9.6 years follow-up is disappointing and merits some re-appraisal of the widely accepted principle that pituitary surgery must be the initial treatment of choice in pituitary-dependent Cushing's syndrome. Following pituitary surgery, careful ongoing expert endocrine assessment is mandatory as the incidence of relapse increases with time and also with increasing rigour of the endocrine evaluation. A significant number of our patients were shown to have relapsed with a cyclical form of hypercortisolism. PMID- 16268809 TI - Maternal hypothyroidism in early and late gestation: effects on neonatal and obstetric outcome. AB - BACKGROUND: Maternal hypothyroidism may be associated with a variety of adverse neonatal and obstetric outcomes. Whether these outcomes are affected by maternal thyroid status at initial presentation or in late gestation specifically within a dedicated antenatal endocrine clinic remains unclear. The effects of thyroxine dose requirement during pregnancy and serum concentrations of TSH within such clinic settings are still not known. OBJECTIVES: We investigated these outcomes in patients with hypothyroidism during early and late gestation. TSH levels and thyroxine dose requirement during early and late gestation were also evaluated. METHODS: We performed a retrospective study of data from 167 pregnancies managed in the antenatal endocrine clinic. Analysis of outcomes was linked to TSH at first presentation and in the third trimester. Outcome variables included: rate of caesarean section, pre-eclampsia, neonatal unit admission, neonatal weight and gestational age. Controlled TSH was defined as mothers with TSH between 0.1 and 2 with normal free thyroid hormone levels. RESULTS: The caesarean section (CS) rates were higher in the study cohort (H) compared with the local (C) rate (H = 28.7%, C = 18%). The higher rate in our patient cohort was not due to a higher rate of emergency section nor to a lower threshold for performing elective caesarean section. The infant birthweight (IBW) from mothers with TSH > 5.5 (H1) and mothers with TSH between 0.1 and 5.5 at presentation (H2) was [median (range)] 3.38 (1.73-4.70) vs. 3.45 (1.36-4.76); P = ns. The prevalence of low birthweight (LBW) infants (< 2.5 g) in groups H1 and H2 was 15% and 4.8%, respectively [odds ratio (OR) = 3.55, 95% confidence interval (95% CI) = 0.96 10.31]. IBW from mothers with TSH > 2 (H3) and mothers with controlled TSH in the third trimester (H4) were similar [3.38 (1.78-4.4) vs. 3.46 (1.36-4.76); P = ns]. The prevalence of LBW in groups H3 and H4 was 9% and 4.9%, respectively (OR = 1.95, 95% CI = 0.52-7.26). The median thyroxine dose (microg) increased significantly during pregnancy (first trimester: 100; second trimester: 125, P < 0.001; and third trimester: 150, P < 0.001) associated with appropriate suppression of TSH levels in the second and third trimesters. Rates of pre eclampsia or admissions to neonatal units were negligible. CONCLUSION: Thyroxine dose requirement increases during pregnancy and thus close monitoring of thyroid function with appropriate adjustment of thyroxine dose to maintain a normal serum TSH level is necessary throughout gestation. Within a joint endocrine-obstetric clinic, maternal hypothyroidism at presentation and in the third trimester may increase the risk of low birthweight and the likelihood for caesarean section. The latter observation was not due to a higher rate of emergency caesarean section nor to a lower threshold for performing elective caesarean section. A larger study with adjustments made for the various confounders is required to confirm this observation. PMID- 16268810 TI - Milk-alkali syndrome is a major cause of hypercalcaemia among non-end-stage renal disease (non-ESRD) inpatients. AB - OBJECTIVE: Milk-alkali syndrome, once a common cause of hypercalcaemia, is now considered rare. Our aim was to estimate the prevalence of milk-alkali syndrome among hypercalcaemic, non-end-stage renal disease (non-ESRD) inpatients of a University Hospital and identify patients' and syndrome characteristics. DESIGN AND PATIENTS: In this retrospective chart review study, we identified patients hospitalized with possible hypercalcaemia between November 1998 and October 2003 by a computer search of admission, discharge and consultation diagnoses. Patients with renal transplantation, stage 5 chronic kidney disease (CKD-5) and those admitted for parathyroidectomy were excluded. The remaining patients' charts were reviewed for confirmation of hypercalcaemia and identification of the cause. In patients with milk-alkali syndrome, additional historical, clinical, laboratory and imaging data were collected. RESULTS: We identified 125 patients with hypercalcaemia, 11 (8.8%) of whom had milk-alkali syndrome, 42 (33.6%) had malignancy and 37 (29.6%) hyperparathyroidism. Thirty-five patients had severe hypercalcaemia, defined as corrected serum calcium 3.5 mmol/l. Malignancy accounted for 13 of those patients (37.1%) and milk-alkali for nine (25.7%). Conditions prevalent among the milk-alkali inpatients were female gender, hypertension, chronic kidney disease, osteoporosis, upper gastrointestinal diseases, diuretic treatment and vitamin D derivative supplementation. Five of the patients with milk-alkali syndrome were treated with bisphosphonates and all five developed hypocalcaemia, compared to one of the five who received conventional treatment (P = 0.047). CONCLUSION: Milk-alkali was the third leading cause of hypercalcaemia of any degree and the second cause of severe hypercalcaemia among inpatients without ESRD. In milk-alkali syndrome, treatment with bisphosphonates contributes to post-treatment hypocalcaemia. PMID- 16268811 TI - Thyroid papillary cancers: microcarcinoma and carcinoma, incidental cancers and non-incidental cancers - are they different diseases? AB - OBJECTIVE: Tumour size represents a much-debated prognostic factor in papillary cancer, and the necessity to perform a fine-needle aspiration (FNA) on small nodules is a frequent matter of discussion. We compared some prognostic histological features for various sizes of papillary cancers (PCs) and, with regard to these prognostic features, we compared non-incidental with incidental PCs. We also considered the possibility that ultrasonography could detect nodules harbouring the most aggressive cancers. DESIGN AND PATIENTS: We have studied patients with a histological diagnosis of PC from 1999 to 2003. FNA was performed on all nodules > 1.0 cm and on hypoechoic nodules with irregular margins or microcalcification when the size was < 1.0 cm (3321 FNAs in total). We were able to consider several different types of patients: those with PC diagnosed by FNA before surgery; those with large goitre and PC of small size diagnosed after histological examination and in whom a careful examination of the presurgical ultrasonography could show a distinct highly suspicious nodule that was not subjected to FNA, and patients with real incidental PC (that is, those with nodular goitre who correctly underwent FNA on suspicious nodules but in whom thyroid cancer was discovered only at histological examination). We considered two groups of patients with PC. Group 1 PCs were diagnosed before surgery with FNA (128 cases); in this group we also considered the two cases that were not correctly diagnosed before surgery. Group 2 real incidental PCs (12 cases) were found in goitres at histological examination after thyroidectomy for goitre (282 thyroidectomies). Group 1 was divided into three subgroups according to the maximum size of the PC at histological examination: (a) 44 PCs with maximum size < 10 mm, (b) 47 PCs with maximum size between 10 mm and 20 mm, and (c) 39 PCs with maximum size 20 mm. In all subgroups 1 as well as in group 2, the following four histological features were considered separately: multifocality, extracapsular extension, lymph-node involvement and its extent, and special aggressive features (dedifferentiation and/or insular aspects, tall and columnar variants). RESULTS: In subgroups 1a, 1b and 1c the results were, respectively: multifocality 56.8, 57.4 and 51.2%; extracapsular extension 27.2, 23.4 and 46.3% (P = 0.01, subgroups 1a and 1b vs. subgroup 1c); lymph-node metastasis 13.5, 23.3 and 46.1% (P = 0.003 subgroup 1a vs. 1c; P = 0.04, subgroup 1b vs. 1c); special aggressive features 11.3, 25.5 and 28.2% (P = ns). Group 2 showed one case of multifocality (8.3%) in a patient with prior exposure to radiotherapy in childhood, while no case was found of extracapsular invasion or lymph-node involvement, and only one patient had a PC with features of dedifferentiation. CONCLUSIONS: Non-incidental cancer, apart from multifocality, showed a classical progression for all prognostic factors from microcarcinoma to larger cancers. However, real incidental PC seemed to be different from non-incidental PC microcarcinoma regarding the main prognostic features. We conclude that ultrasonography is useful not only in terms of revealing the presence of cancer but also in identifying the most aggressive cancers. PMID- 16268812 TI - Serum biomarkers for cardiovascular inflammation in subclinical hypothyroidism. AB - OBJECTIVE: Cardiovascular disease has been reported to be more common in patients with elevated levels of TSH and with normal thyroxine levels, termed subclinical hypothyroidism (SCH). The aim of this study was to determine whether individuals with SCH were more likely than euthyroid controls to have elevations in inflammatory biomarkers that are associated with cardiac disease. METHODS: From the National Health and Nutrition Examination Survey (NHANES) 1999-2002 data we identified respondents aged > or = 40 who met the laboratory criteria for SCH. We compared the average values and frequencies of elevated C-reactive protein (CRP) and homocysteine in the SCH individuals to those in euthyroid individuals. Results A total of 1608 individuals were included in the analysis of whom 45 (2.8%) met the criteria for SCH, with seven (15.6%) having TSH levels between 10 and 15 IU/l. We found no differences in median, mean or the percentage of elevated CRP or homocysteine in individuals with SCH compared to euthyroid controls. Linear regression models adjusting for age, gender, the presence of other cardiac risk factors, and whether the individual was using a cholesterol lowering drug also failed to show any association between SCH and either high sensitivity CRP (hsCRP) (P = 0.62) or homocysteine (P = 0.63) levels. When we examined the subset of people with TSH levels between 10 and 15 IU/l, we again found no difference in mean hsCRP or homocysteine compared to control. CONCLUSIONS: In this large population-based sample, hsCRP and homocysteine levels do not differ for individuals with SCH compared to euthyroid individuals. This suggests that cardiac inflammation is not greater in individuals with SCH. PMID- 16268813 TI - The BRAF mutation is not associated with poor prognostic factors in Korean patients with conventional papillary thyroid microcarcinoma. AB - BACKGROUND: The BRAF(V600E) mutation, the most common genetic alteration reported in papillary thyroid carcinoma, has been associated with poor prognostic factors. AIM: To determine whether the presence of the BRAF(V600E) mutation is associated with poor prognosis in Korean patients with conventional papillary thyroid microcarcinoma (micro-PTC). PATIENTS AND METHODS: DNA was extracted from paraffin embedded thyroid tumour specimens taken from 60 patients with conventional micro PTC, as well as from nine patients with follicular variant papillary carcinoma, six with nodular hyperplasia, four with follicular carcinoma (including one with Hurthle cell carcinoma), four with follicular adenoma (including two with Hurthle cell adenoma) and one each with medullary carcinoma, poorly differentiated carcinoma and anaplastic carcinoma. The presence of the BRAF(V600E) mutation was determined by polymerase chain reaction (PCR) amplification of exon 15 followed by direct sequencing. RESULTS: The BRAF(V600E) mutation was detected in tumour samples from 31 of 60 conventional micro-PTC patients (52%), but was not detected in patients with other types of thyroid tumours. The age distribution, tumour size, extrathyroid extension, multifocality and staging did not differ significantly between patients with and without the BRAF(V600E) mutation. CONCLUSION: In Korean patients with conventional micro-PTC, the presence of the BRAF(V600E) mutation was not significantly associated with prognostic factors. PMID- 16268814 TI - Serum osteoprotegerin levels are associated with inflammation and pulse wave velocity. AB - OBJECTIVE: We examined the association between serum osteoprotegerin (OPG) levels, systemic inflammation and arterial stiffness in normal and diabetic patients. PATIENTS AND MEASUREMENTS: The study subjects comprised 49 newly diagnosed diabetic patients and 72 age- and sex-matched normal glucose controls. Anthropometric parameters, blood pressure, fasting blood glucose (FBG), lipid profiles, serum OPG, high-sensitive C-reactive protein (hsCRP), interleukin-6 (IL 6) and brachial-ankle pulse wave velocity (baPWV) were measured. RESULTS: Serum OPG levels (6.1 +/- 1.4 vs. 5.4 +/- 1.3 pmol/l, P = 0.011) and baPWV (1562 +/- 354 vs. 1399 +/- 257 cm/s, P = 0.004) were significantly higher in the diabetic group than in the normal glucose group. Serum OPG levels in normal and diabetic patients correlated significantly with systolic blood pressure (r = 0.20, P = 0.035), FBG (r = 0.30, P = 0.002), right baPWV (r = 0.22, P = 0.021), left baPWV (r = 0.26, P = 0.006), homeostasis model assessment insulin resistance (HOMA-IR) (r = 0.19, P = 0.045), IL-6 (r = 0.32, P = 0.001) and hsCRP (r = 0.21, P = 0.027) after adjusting for age and sex. Multiple regression analysis showed that serum OPG level was significantly associated with age, FBG, IL-6, systolic blood pressure, triglyceride and hsCRP (R(2) = 0.299). CONCLUSIONS: In summary, serum OPG and baPWV levels are elevated in diabetic patients and serum OPG levels are significantly associated with inflammation and arterial stiffness. PMID- 16268816 TI - Serological markers of autoimmunity in patients infected with hepatitis C virus: impact of HIV co-infection. AB - OBJECTIVES: We sought to evaluate the prevalence, predictors and significance of autoantibody expression in patients with chronic hepatitis C (CHC) with or without HIV co-infection. METHODS: Retrospective review of laboratory and histologic data for all patients with CHC who had a liver biopsy available. HIV status was documented in all patients. Results analyzed in SPSS10, Chicago, IL, a p value <0.05 was considered significant. RESULTS: 170 patients with hepatitis C viremia, including 107 (63%) HIV co-infection, who had testing for anti-nuclear antibody (ANA) or anti-smooth muscle antibody (ASMA) and anti-mitochondrial antibody (AMA) were included in the study. Overall, 63% (74/117) of patients were ASMA seropositive and 6% (9/153) were positive for ANA. All 117 patients tested for AMA were negative. HIV co-infected patients were significantly more likely to be ASMA positive 71% (53/75) compared to those with hepatitis C alone (50%) [P=0.026]. There were no significant differences in age, gender, race, risk group, alanine aminotransferase (ALT) levels or grade of inflammation on histology between autoantibody positive and negative patients. ASMA positive patients had significantly higher globulin levels (P=0.036) and a trend towards more bridging fibrosis or cirrhosis. Patients with autoantibody expression rarely had histologic features of AIH. CONCLUSION: We found a high rate of ASMA seropositivity in our cohort of patients with chronic hepatitis C, and HIV co infection was associated with significantly higher rates of ASMA expression. Autoantibody expression was not associated with demographic or clinical characteristics and does not necessarily preclude antiviral therapy. PMID- 16268817 TI - Validation of a simple model for predicting liver fibrosis in HIV/hepatitis C virus-coinfected patients. AB - OBJECTIVES: Recently, several models incorporating laboratory measurements have been validated for use as surrogate markers for liver fibrosis in hepatitis C virus (HCV) mono-infection, the simplest of these being the aspartate aminotransferase (AST) to platelet ratio index (APRI). We evaluated how well the APRI predicts significant hepatic fibrosis in patients with HIV/HCV coinfection. METHODS: Forty-six HIV/HCV-coinfected patients who underwent liver biopsy and had concomitant laboratory measurements (+/-3 months) were included in the study. Significant fibrosis was defined as F2-F4 using Batt and Ludwig scoring (=3 Ishak). APRI=[(AST/upper limit of normal)/platelet count (10(9)/L)] x 100. We used sas proc logistic (SAS Institute, Cary, NC) to calculate the area under the receiver operating curve (ROC) (AUC). Sensitivities, specificities, positive predictive value (PPV) and negative predictive value (NPV) were compared using cut-offs previously identified in the literature. RESULTS: Thirty-three of 46 patients (72%) had significant fibrosis on biopsy. For significant fibrosis, the area under the ROC for the APRI was 0.847+/-0.057. APRI scores >1.5 (the higher cut-off) were 100% specific and 52% sensitive; PPV was 100% and NPV 45%. Scores <0.5 (the lower cut-off) were 82% sensitive and 46% specific in ruling out significant fibrosis (PPV 79%; NPV 50%). CONCLUSIONS: A simple model incorporating readily available laboratory data is highly predictive of significant fibrosis in HIV/HCV coinfection and could serve as a biopsy-sparing measure, thus making treatment more accessible for this population. PMID- 16268818 TI - Rash in an HIV-positive patient. PMID- 16268819 TI - Carotid intima-media thickness is slightly increased over time in HIV-1-infected patients. AB - OBJECTIVES: HIV-infected patients are at risk of atherosclerosis and cardiovascular diseases. In a 12-month follow-up study, we aimed to investigate changes in carotid intima-media thickness (IMT), a surrogate marker of atherosclerosis, and its determinants in HIV-1-infected patients. METHODS: Our multicentre prospective longitudinal cohort study included 346 HIV-infected patients, for each of whom two IMT measurements were taken by B-mode ultrasonography at baseline (M0) and 1 year later (M12). RESULTS: We observed a significant but moderate increase in the common carotid artery (CCA) median IMT, from 0.54 to 0.56 mm (P<10(-4)), i.e. an increase of 0.020 mm (95% confidence interval 0.012-0.029). There was a significant association between cross sectional CCA IMT measures at M12 and conventional cardiovascular risk factors (higher CCA IMT with older age, P<10(-4); male gender, P=0.02; tobacco consumption, P=0.05), as well as higher CD4 cell count at M12 (>median 455 cells/microL, P=0.01). Only CD4 cell count at M0 was strongly and positively associated with the variation in IMT between M0 and M12 (P=4 x 10(-3)). IMT progression was +0.0020 mm for the lowest quartile of CD4 cell count distribution at M0, i.e. 3-253 cells/microL, +0.010 mm for 253-402 cells/microL, +0.043 mm for 402-590 cells/microL, and +0.028 mm for 590-2270 cells/microL. No association was found with type or duration of antiretroviral exposure. CONCLUSIONS: Conventional cardiovascular risk factors are major determinants of IMT evolution. The link between immunological status and carotid IMT requires further study. PMID- 16268820 TI - Triple nucleoside reverse transcriptase inhibitor- vs. nonnucleoside reverse transcriptase inhibitor-containing regimens as first-line therapy: efficacy and durability in a prospective cohort of French HIV-infected patients. AB - OBJECTIVE: Based on the short-term results of the AIDS Clinical Trials Group (ACTG) A5095 trial, zidovudine (ZDV)/lamivudine (3TC)/abacavir (ABC) is no longer recommended as a first-line antiretroviral regimen. Data on the efficacy of this triple nucleoside reverse transcriptase inhibitor (NRTI) combination compared with the gold-standard nonnucleoside reverse transcriptase inhibitor (NNRTI) regimen could provide important information. METHODS: Patients were selected from three prospective cohorts of patients who received first-line therapy with ZDV/3TC plus an NNRTI or ABC, started after January 1998. Immunovirological changes and the proportion of treatment discontinuations were compared between groups. RESULTS: Of the 380 patients, 190 started on ABC [the triple-NRTI group (3N)] and 190 on NNRTI. At baseline, there was no statistical difference between the NNRTI and 3N groups for age (mean=38 years), sex (66% male) or CD4 cell count (mean=305 cells/muL). Mean baseline plasma HIV-1 viral load (pVL) was higher in the 3N group (4.6 vs. 4.3 log10 HIV-1 RNA copies/mL: P<0.01). Lower and higher estimates of median pVL decrease at month 24 were 2.05 and 4.76 log10 copies/mL in the 3N group, and 1.73 and 4.31 log10 copies/mL in the NNRTI group (not significant). CD4 cell count evolution did not differ between groups. Treatment discontinuation occurred in 45% vs. 44% of patients in the NNRTI and 3N groups, respectively, after median durations of 9 and 4 months, respectively (P=0.02). CONCLUSIONS: In this prospective cohort, 3N and NNRTI regimens as first-line therapy produced similar immunovirological responses. PMID- 16268821 TI - Impact of antiretroviral choice on hypercholesterolaemia events: the role of the nucleoside reverse transcriptase inhibitor backbone. AB - BACKGROUND: The use of highly active antiretroviral therapy (HAART) has profoundly altered the life expectancy of individuals infected with HIV. Metabolic abnormalities associated with antiretrovirals and cumulative exposure to combination antiretroviral therapy, including dyslipidaemia and insulin resistance, have been linked to an increased risk of myocardial infarction. METHODS: Longitudinal data from a large prospectively collected clinical database were analysed. All patients who commenced first antiretroviral therapy (ART) [two nucleoside reverse transcriptase inhibitors (NRTIs)+one nonnucleoside reverse transcriptase inhibitor (NNRTI) or one active protease inhibitor (PI)] since 1996 were identified. Patients with elevated cholesterol levels [>5.5 mmol/L (215 mg/dL)] prior to therapy initiation were excluded. Quantitative data were categorized into quartiles and presented stratified by individuals developing abnormal levels of cholesterol during first-line HAART. Event time was defined as time from commencing first-line ART to either development of cholesterol level >6.5 mmol/L (254 mg/dL) or switch of first-line therapy. The Kaplan-Meier product limit survival method was used to estimate time to abnormal cholesterol level, and the chi2 test was used for comparisons between drug classes. Cox's proportional hazards regression analysis was used to identify factors predicting a likelihood of raised cholesterol level. RESULTS: A total of 1664 patients were included in the study: 57.1% on two NRTIs+one NNRTI, 38.4% on two NRTIs+one PI, and 4.4% on two NRTIs+a boosted PI regimen. Regimens containing stavudine or PIs were associated with a significantly higher event risk and earlier time to event. No differences between efavirenz and nevirapine or between didanosine and lamivudine were observed. In 28 patients exposed to the combination of tenofovir+lamivudine+efavirenz, there were no episodes of elevated cholesterol level. CONCLUSION: Dyslipidaemia has emerged as an important issue in HIV infected individuals receiving antiretroviral therapy. This study demonstrates that age at start of therapy, baseline cholesterol level, stavudine use and PI use are all associated with increased risk of hypercholesterolemia on initial therapy. Both NRTI and NNRTI/PI choice influence risk of hypercholesterolaemia. PMID- 16268822 TI - Drug-resistance mutations in antiretroviral-naive patients with established HIV-1 infection in Mexico. AB - OBJECTIVES: To describe the prevalence of baseline drug-resistance mutations, resistance to antiretroviral drugs, and the subsequent virological response to therapy in treatment-naive patients from Mexico with established HIV-1 infection. METHODS: Resistance testing was performed on plasma samples from antiretroviral naive patients. Data on mutations associated with antiretroviral drug resistance were obtained using Stanford software (http://hivdb.stanford.edu). RESULTS: Ninety-six treatment-naive individuals were enrolled in the study during 2002 2003. Of these, 83 patients (86%) had at least one resistance mutation and 15 (16%) had drug resistance. At baseline, the mean plasma viral load was 299 834 HIV-1 RNA copies/mL, and at follow-up it was 37 620 copies/mL (P<0.0001). Primary mutations in the reverse transcriptase region were observed in 15% of patients. For nucleoside inhibitors, mutations T215Y/C and F77L (3%) and D67N/S, T69N and M184V (2%), were detected. For nonnucleoside inhibitors, mutations K103N/R (6%), Y181C (3%) and G190A (2%) were detected. Overall, 6% of patients showed resistance to delavirdine and nevirapine, 4% to efavirenz, and 2% to lamivudine and nelfinavir. Twelve patients showed no response to treatment and three of these patients had antiretroviral drug resistance. CONCLUSIONS: The prevalence of baseline drug-resistance mutations found in this study was similar to that found in previous reports for newly HIV-infected individuals, although access to and management of antiretrovirals in Mexico are different. PMID- 16268823 TI - Indinavir/ritonavir 800/100 mg bid and efavirenz 600 mg qd in patients failing treatment with combination nucleoside reverse transcriptase inhibitors: 96-week outcomes of HIV-NAT 009. AB - OBJECTIVE: Nucleoside reverse transcriptase (NRTI) sparing is a favourable option for patients with NRTI failure or toxicity. METHODS: Patients judged to be failing NRTI therapy were enrolled in a single-arm, open-label study of indinavir/ritonavir (IDV/r) 800/100 mg twice a day (bid)+efavirenz (EFV) 600 mg once a day (qd). The primary endpoint was the change in time-weighted average HIV RNA from baseline. The initial 48-week protocol was extended to 96 weeks by a single amendment. Analysis was by intention to treat. RESULTS: Sixty-one patients (23 female) were enrolled in the study. Baseline median inter-quartile range (IQR) NRTI exposure was 4.4 (3.9-4.7) years; baseline median viral load was 4.09 log(10) HIV-1 RNA copies/mL (range 3.75-4.61 log(10) copies/mL); baseline median CD4 count was 169 cells/microL (range 60-277 cells/microL). The mean (SD) change in time-weighted average HIV RNA from baseline at 48 and 96 weeks was -2.1 (0.7) and -2.1 (0.8) log(10) copies/mL respectively, resulting in 87% and 69% of patients with HIV RNA <50 copies/mL. Sixteen per cent of patients permanently ceased therapy and 26% underwent temporary drug interruptions because of study drug-related adverse events. Fasted-lipid values rose significantly over the 96 weeks of study, as did median blood glucose and median serum creatinine levels. Twelve (20%) patients underwent IDV dose reduction, mainly because of nephrotoxicity (nine of 12 patients). Blood pressure values deteriorated following switch, but markers of nucleoside toxicity improved. CONCLUSIONS: IDV/r 800/100 mg bid+EFV 600 mg qd gave a potent, durable response in these NRTI failures and was reasonably well tolerated. However, we observed adverse effects on renal, metabolic and blood pressure parameters. Lower doses of boosted IDV might improve toxicity while maintaining efficacy, and this possibility warrants further investigation. PMID- 16268824 TI - The effect of low-dose ritonavir monotherapy on fasting serum lipid concentrations. AB - OBJECTIVES: Ritonavir (RTV) at doses of 400 mg twice a day (bid) or higher adversely affects serum lipids. However, the effect of RTV 100 mg bid on serum lipids is unknown. We conducted a study to evaluate the effect of RTV 100 mg bid on fasting serum lipid profiles in HIV-negative healthy volunteers. METHODS: Ritonavir 100 mg bid was administered for 14 days to 20 healthy HIV-seronegative adults with normal serum lipids. After a 7-day washout, lopinavir/ritonavir (LPV/RTV) 400/100 mg bid was administered for 14 days. Fasting serum lipid parameters were measured twice at baseline, after 14 days of RTV, and after 14 days of LPV/RTV, and comparisons were made at each time-point for levels of total cholesterol, low-density lipoprotein (LDL) cholesterol, high-density lipoprotein (HDL) cholesterol, the total/HDL cholesterol ratio and triglycerides. RESULTS: After 14 days of RTV 100 mg bid, total cholesterol level increased by 10.2% (P<0.001), LDL cholesterol level increased by 16.2% (P<0.001), triglyceride levels increased by 26.5% (P<0.001), HDL cholesterol level decreased by 5.4% (P<0.01) and the total/HDL cholesterol ratio increased by 17.3% (P<0.001). The addition of LPV 400 mg bid to RTV 100 mg bid resulted in no significant further changes in LDL cholesterol or triglyceride level or total/HDL cholesterol ratio, but there were significant increases in both total cholesterol (8.0% increase; P=0.007) and HDL cholesterol levels (6.7% increase; P=0.008). CONCLUSIONS: Ritonavir dosed at 100 mg bid significantly increased the concentration of total cholesterol, LDL cholesterol, total/HDL cholesterol ratio and triglycerides and reduced HDL cholesterol concentration. The addition of LPV 400 mg bid to RTV 100 mg bid further increased both total and HDL cholesterol levels without affecting the total/HDL ratio. PMID- 16268827 TI - Measuring research productivity. PMID- 16268828 TI - Interaction between ozone and airborne particulate matter in office air. AB - This study investigated the hypotheses that humans are affected by air pollution caused by ozone and house dust, that the effect of simultaneous exposure to ozone and dust in the air is larger than the effect of these two pollutants individually, and that the effects can be measured as release of cytokines and changes of the respiratory function. Experimental exposures of eight atopic but otherwise healthy subjects were performed in a climate chamber under controlled conditions. The three controlled exposures were about 75 microg/m3 total suspended particulate matter, 0.3 p.p.m. ozone, and the combination of these. The exposure duration was 3 h. The outcome measures were interleukins and cells in nasal lavages (NAL), respiratory function, bronchial metacholine responsiveness, rhinometry symptoms and general well-being in a questionnaire and time course of general irritation on a visual analogue scale. Indications of interactions between exposure types were demonstrated for peak expiratory flow (PEF) (P<0.05) and for discomfort symptoms (P<0.03). Non-significant interactions were found for the concentration of interleukin-8 in NAL. The combined exposure was found to cause significantly more effects than either ozone exposures or dust exposures. This is interpreted as indications of a potentiation caused by the combined exposures to dust and ozone. The findings in this study are based on a limited number of subjects and thus should not be over-interpreted. However, they support the hypothesis that ozone at relatively high concentrations interacts with dust exposures to cause decrements in PEF and increase in discomfort measures. PRACTICAL IMPLICATIONS: If confirmed at lower ozone and dust concentrations this finding could help to explain many problems with indoor air quality reported in offices throughout the world. PMID- 16268829 TI - Ventilation in public housing: implications for indoor nitrogen dioxide concentrations. AB - Although elevated nitrogen dioxide (NO2) exposures may exacerbate asthma, few studies have examined indoor NO2 levels in low-income, urban neighborhoods, where asthma prevalence is high. As part of the Healthy Public Housing Initiative, NO2 was measured in 77 homes within three Boston public housing developments, using Palmes tubes placed in the kitchen, living room, and outdoors. Air exchange rates (AERs) were assessed using a perfluorocarbon tracer technique. Overall NO2 levels were [mean (ppb)+/-s.d.]: kitchen (43+/-20, n=100), living room (36+/-17, n=102), outdoor (19+/-6, n=91). Indoor NO2 levels were significantly higher in the heating season (living room: 43 ppb vs. 26 ppb, kitchen: 50 ppb vs. 33 ppb), while AERs were significantly lower in the heating season (medians 0.49/h vs. 0.85/h). Significant univariate predictors of indoor concentrations include: outdoor NO2 levels, AERs, and occupancy. AERs and outdoor NO2 remained significant in multivariate models (P<0.05). A dummy variable for supplemental heating with gas stove was not significant (P=0.14), but had a large, positive coefficient. Indoor NO2 levels in this cohort are higher than those generally reported in residential US settings, associated in part with increased gas stove usage and decreased AERs during the heating season. PRACTICAL IMPLICATIONS: Indoor air quality is mainly a function of outdoor concentrations, indoor sources, ventilation, and residential behavior. Indoor exposures to nitrogen dioxide and other combustion pollutants may be elevated within low-income housing developments due to the presence of multiple sources, poor ventilation, small apartment size, and behavioral responses to apartment conditions (e.g. supplemental heating with gas stove). This information may be used by housing authorities and other landlords to decrease potential environmental stressors, through interventions such as source substitution and improved ventilation, particularly for sensitive sub-populations such as asthmatics. PMID- 16268830 TI - A pilot study to investigate the effects of combined dehumidification and HEPA filtration on dew point and airborne mold spore counts in day care centers. AB - Meteorological factors such as relative humidity directly correlate with airborne fungal levels outdoors and indoors. While dehumidification alone is effective at reducing moisture necessary for mold growth, it is inadequate as a single intervention as it does not remove viable and non-viable fungal spores that are potentially allergenic. The purpose of this pilot study was to investigate whether dehumidification in combination with high-efficiency particulate arrestance (HEPA) filtration is effective at reducing airborne mold spore levels in day care centers. Two day care centers within a 2-mile radius of each other were selected. Day care center A was 2 years old with eight rooms while day care center B was 15 years old with six rooms. A high efficiency Santa Fe dehumidification unit equipped with a HEPA filter was installed in half the rooms (intervention) of each day care facility. Electronic HOBO data loggers continuously measured outdoor and indoor room dew point and temperature every 2 h throughout the study. Dew point and airborne fungal spore measurements from selected rooms with controlled air conditions were analyzed by comparing baseline measurements to those obtained at subsequent time periods over 1 year. Regression models accounted for correlations between measurements in the same room over time. Intervention resulted in a lowered average dew point from baseline by 8.8 degrees C compared with a decrease of 7.1 degrees C in non-intervention rooms across all time periods in both facilities (P<0.001). Fungal analyses demonstrated lower baseline (P=0.06) and follow-up means in intervention rooms (P<0.05), however the change from baseline to end of follow-up differed between intervention and non-intervention rooms in the two facilities. Log transformation was applied to approximate normality of fungal measurements. Dehumidification with HEPA filtration was effective at controlling indoor dew point in both facilities and at reducing airborne culturable fungal spore levels in one of the two facilities. These preliminary results provide a scientific rationale for using this intervention in future studies designed to investigate the impact of indoor mold exposure on health outcomes. PRACTICAL IMPLICATIONS: Poor indoor air quality is a recognized cause or contributing factor to health effects. Dampness and humidity have been linked to upper and lower respiratory symptoms in children and adults. This study indicates that reducing indoor relative humidity and airborne mold spore levels using high-efficiency dehumidification units equipped with HEPA filtration is feasible even in work facilities such as day care centers where traffic in and out of the building is difficult to regulate. Clinicians should emphasize to their patients the importance of dehumidification and HEPA filtration to improve indoor air quality in the home and workplace. PMID- 16268831 TI - The influence of ammonia and carbon dioxide on the sorption of a basic organic pollutant to a mineral surface. AB - Indoor surfaces have a sorptive capacity for organic pollutants which may be significantly influenced by other gases and the pH of the surface. In this research, we examine the influence of a common indoor gaseous acid, CO2, and base, NH3, on the adsorption of a volatile organic base, trimethylamine (TMA), to a mineral surface, zirconium silicate beads. Varying ammonia and CO2 within concentration ranges of indoor relevance substantially influences the sorptive capacity of this mineral surface. Increasing the CO2 mixing ratio to 1000 p.p.m. enhances surface capacity of TMA by 40-50%; increasing the NH3 mixing ratio to 10 p.p.m. decreases the TMA surface capacity by approximately 5-80% depending on relative humidity. The phenomena of dissolution of TMA into bulk surface water and acid-base chemistry in the surface water do not adequately describe equilibrium adsorption on this surface. Instead, adsorption to the dry solid or to adsorbed water layers appears to dominate. Reduction in the equilibrium partition coefficient, ke, in the presence of NH3 is due to a competition between TMA and ammonia molecules for adsorption sites. Site competition appears to follow the Langmuir competitive model and most ke values range from 0.003-0.045 m. PRACTICAL IMPLICATIONS: Sorptive interactions with indoor surfaces strongly influence indoor exposure to pollutants. For basic or acidic compounds, these interactions are themselves influenced by surface pH and competition with other acidic or basic gases such as CO2 and NH3. We show that CO2 tends to cause mineral surfaces to store more amines but NH3 tends to decrease this surface capacity. Given the typical range of indoor CO2 and NH3 concentrations, the indoor sorbtive capacity of amines on mineral surfaces may vary by greater than an order of magnitude. PMID- 16268832 TI - Use of air-cleaning devices to create airborne particle-free spaces intended to alleviate allergic rhinitis and asthma during sleep. AB - Comprehensive quantitative experiments were performed to assess the capabilities of several air-cleaning devices to create a particle-free microenvironment as a therapy for sleeping persons affected by allergic rhinitis and asthma. Six devices were evaluated, of which five were portable and intended to provide general air cleaning for bedroom-sized spaces. The sixth was intended for installation in front of the headboard of a bed and was designed to provide clean air focused in a space occupied by a sleeping person. The air-cleaning methods of the selected devices included high-efficiency particulate air (HEPA) filtering and electrostatic precipitation. Particle concentration measurements for six particle-size ranges and sound intensity measurements were made during 8-h, sleep simulating periods. The effects of four parameters were studied: (i) device location, (ii) controlled air motion in the laboratory, (iii) airflow rate setting of the air-cleaning device, and (iv) controlled disturbances. To ensure a totally objective study, a special laboratory facility was constructed which enabled complete control of the experimental conditions. The measured concentration histories provided comprehensive evidence of the relative capabilities of the various devices for the specific air-cleaning function. It was found that the device designed to focus the cleaned air in the sleeping space fulfilled its goal and, in that regard, was clearly superior to all of the other air-cleaning devices. PRACTICAL IMPLICATIONS: There is evidence that allergic reactions are triggered by the presence of airborne particles and that these reactions can be mitigated by particle removal. This strategy can be implemented by the use of air-cleaning devices which are capable of creating particle-free zones at locations where human activity occurs. In particular, the creation of a particle-free zone which encompasses the breathing space of a sleeping person holds promise of mitigating sleep-disturbing allergic reactions. PMID- 16268833 TI - Effects of an ozone-generating air purifier on indoor secondary particles in three residential dwellings. AB - The use of indoor ozone generators as air purifiers has steadily increased over the past decade. Many ozone generators are marketed to consumers for their ability to eliminate odors and microbial agents and to improve health. In addition to the harmful effects of ozone, recent studies have shown that heterogeneous and homogeneous reactions between ozone and some unsaturated hydrocarbons can be an important source of indoor secondary pollutants, including free radicals, carbonyls, carboxylic acids, and fine particles. Experiments were conducted in one apartment and two detached single-family dwellings in Austin, TX, to assess the effects of an ozone generator on indoor secondary organic aerosol concentrations in actual residential settings. Ozone was generated using a commercial ozone generator marketed as an air purifier, and particle measurements were recorded before, during, and after the release of terpenes from a pine oil-based cleaning product. Particle number concentration, ozone concentration, and air exchange rate were measured during each experiment. Particle number and mass concentrations increased when both terpenes and ozone were present at elevated levels. Experimental results indicate that ozone generators in the presence of terpene sources facilitate the growth of indoor fine particles in residential indoor atmospheres. Human exposure to secondary organic particles can be reduced by minimizing the intentional release of ozone, particularly in the presence of terpene sources. PRACTICAL IMPLICATIONS: Past studies have shown that ozone-initiated indoor chemistry can lead to elevated concentrations of fine particulate matter, but have generally been completed in controlled laboratory environments and office buildings. We explored the effects of an explicit ozone generator marketed as an air purifier on the formation of secondary organic aerosol mass in actual residential indoor settings. Results indicate significant increases in number and mass concentrations for particles <0.7 microns in diameter, particularly when an ozone generator is used in the presence of a terpene source such as a pine oil-based cleaner. These results add evidence to the potentially harmful effects of ozone generation in residential environments. PMID- 16268834 TI - Odor and chemesthesis from brief exposures to TXIB. AB - An experiment explored ability of subjects to detect vapors of the plasticizer TXIB (2,2,4-trimethyl-1,3-pentanediol diisobutyrate) and ethanol via olfaction and via ocular and nasal chemesthesis, i.e. chemically stimulated feel. Testing, tailored to the sensitivity of each subject, produced psychometric functions for individuals. Olfactory detection of TXIB began at concentrations below 1 ppb (v/v), with 50% correct detection at 1.2 ppb. (Comparable detection for ethanol occurred almost two orders of magnitude higher.) Chemesthetic detection of TXIB began at about 500 ppb, with 50% correct detection at 2.1 ppm for the eye and 4.6 ppm for the nose, both close to saturated vapor concentration. (Comparable detection for ethanol occurred essentially three orders of magnitude higher.) Suggestions that TXIB plays a role in generation of irritative symptoms at concentrations in the range of parts-per-billion need to reckon with a conservatively estimated 200-fold gap between the levels putatively 'responsible' for the symptoms and those even minimally detectable via chemesthesis. Neither the variable of exposure duration nor that of mixing offers a likely explanation. Inclusion of ethanol in the study allowed comparisons pertinent to issues of variability in human chemoreception. An interpretation of the psychometric functions for individuals across materials and perceptual continua led to the conclusion that use of concentration as the metric of detection in olfaction inflates individual differences. PRACTICAL IMPLICATIONS: This study indicated that the plasticizer TXIB could contribute odor at concentrations in the range of parts-per-billion, but could hardly contribute sensory irritation per se, as alleged in reports of some field studies where TXIB has existed amongst many other organic compounds. PMID- 16268835 TI - A model predicting the effect of speech of varying intelligibility on work performance. AB - Speech is the most distracting sound in (open-plan) offices. Several laboratory studies have shown that speech impairs the performance of, for example, reading and short-term memory. It is not the sound level of speech that determines its distracting power but its intelligibility, which can be physically determined by measuring the Speech Transmission Index (STI). The aim of this study was to develop a mathematical model that predicts how much the performance is reduced due to speech of varying intelligibility. The model was based on the literature according to which performance decrements have been 4-45% depending on the task. The best performance occurs when speech is absent (STI=0.0), and the strongest performance decrement occurs when speech is perfectly heard (STI=1.0). The shape of the performance vs. STI between 0.0 and 1.0 was adopted from the general speech intelligibility theory. The performance starts to decrease when STI exceeds 0.2. Highest performance decrease is reached already when STI exceeds 0.60. PRACTICAL IMPLICATIONS: The prediction model can be exploited in the evaluation of work performance in different acoustical conditions in open-plan offices when STI is known. It can be utilized to promote actions aiming at better acoustical conditions. PMID- 16268837 TI - Higher education for lower patient risk? PMID- 16268836 TI - Risk factors for mold in housing: a national survey. AB - A national random telephone survey was undertaken to determine the prevalence of reported mold in New Zealand houses and the risk factors for it. A total of 613 households provided responses. Mold in one or more rooms was reported by 35.1% of respondents in the sample. House design and construction factors that were independently associated with reported mold in the multivariate analysis included: poorer house condition, older house age (>22 years), relative lack of sun exposure, and having no insulation (e.g. for poorer house condition: odds ratio=1.97, 95% CI=1.25, 3.11). Univariate analyses also showed increased risk associated with high locality rainfall, and living in the most northern part of the country. The number of residents was significantly associated with reported mold in the multivariate analysis as were various behaviors in the univariate analysis (i.e. frequency of baths, showering and clothes washing). The high prevalence of unflued gas heating (32.9%) found in this sample is of potential concern given the potential respiratory hazards. Although this survey has a number of limitations, it does suggest that there are a number of potentially modifiable risk factors for mold that could be reduced by a range of policy responses. PRACTICAL IMPLICATIONS: While further research is desirable, the available evidence associated with the health and other adverse impacts of both mold and dampness would favor additional policy responses by government. These could include changes to regulations around housing design including house position, access to sunlight, and level of insulation. The extension of low interest loans for insulation or subsidized installation can help to increase its level of use. Restrictions on the sales of unflued gas heaters could also be considered. A mass media campaign could also be used to inform the population of the readily modifiable risk factors for mold growth and dampness. PMID- 16268838 TI - Ways of talking about experiences of pain among older patients following orthopaedic surgery. AB - AIM: The aim of this study was to examine how older patients who had undergone hip surgery described their experience of pain. BACKGROUND: A verbal report of pain is considered to be the single most reliable indicator of a person's pain experience. When assessing pain, healthcare professionals must be able to interpret the content of pain reports in order to understand older patient's pain experiences. METHODS: The study was carried out in two orthopaedic and two elder care wards in a large university hospital in Sweden in 2000. Altogether, 38 patients with hip replacement (mean age = 75) and 22 patients with hip fracture (mean age = 81) took part. A face-to-face interview was conducted with each patient on the second day after operation. Data were transcribed and analysed using descriptive qualitative content analysis. FINDINGS: Participants expressed their pain in a nuanced and detailed way in everyday language. Four main themes with sub-themes emerged: (a) objectification (localizing; quantifying; characterizing; temporalizing); (b) compensating (substitution; picturing); (c) explaining (functionalizing pain and its relief; externalizing pain and its relief); (d) existentializing (present pain orientation; future pain orientation). CONCLUSIONS: Exploring the ways older patients talk about pain is expected to result in a better understanding of the older patient's need of empathic individualized care and in the optimization of pain management. PMID- 16268840 TI - A realist study of the mechanisms of cardiac rehabilitation. AB - AIM: The aim of this paper is to report patients' experiences of cardiac rehabilitation and perceptions of the mechanisms and contexts influencing its long-term effectiveness. BACKGROUND: Cardiac rehabilitation programmes for the secondary prevention of coronary heart disease are common. The effects of these programmes, however, can be inconsistent and little is known of the personal and contextual factors that influence service effectiveness. METHOD: Forty-seven participants with a formal diagnosis of coronary heart disease who had attended a programme of cardiac rehabilitation in Scotland 3 years previously were included in focus groups to discuss their perceptions and experiences (30 males and 17 females). The data were generated in 2002 and analysed using the realist approach of Pawson and Tilley (1997). RESULTS: Participants' accounts indicated that the didactic content of cardiac rehabilitation was not strongly linked to longer-term health behaviour change. The main positive effects of cardiac rehabilitation were related to the effect of participation on mediating social and body-focused mechanisms that were triggered when the rehabilitation setting was perceived to be safe. Social mechanisms identified included social comparisons, camaraderie, and social capital. Body-focused mechanisms included greater knowledge of personal physical boundaries and a greater trust in the heart-diseased body. Collectively, these mechanisms had a positive effect on confidence that was perceived as being imperative to maintain health behaviour change. CONCLUSIONS: More support is required to promote health behaviour change after the completion of cardiac rehabilitation. Use of community-based exercise services and conventional or web-based support groups for coronary heart disease patients should be encouraged, as these appear to extend the positive health effects of the mechanisms that promote behaviour change. At the completion of cardiac rehabilitation programmes, patients should be referred to safe and appropriate community-based exercise services. Further research is needed to examine the effects on health outcomes of mechanisms and contexts related to cardiac rehabilitation. PMID- 16268841 TI - Women's experiences of fatigue in chronic illness. AB - AIM: The aim of this paper is to add a qualitative dimension to the body of knowledge about fatigue by revealing the meaning given by women living with chronic illness to the experience of fatigue. BACKGROUND: Fatigue is a common symptom experienced by people who live with chronic illness. It pervades every aspect of life and may be experienced in physical, psychological, emotional or social dimensions. Management of fatigue relies heavily on the individual's ability to employ self-care actions. The invisibility of fatigue is recognized as one of the most frustrating aspects, which can lead to lack of understanding and misunderstanding by others. METHOD: We report the findings of data from research in progress (2003-2005). Data were generated via email group conversations between us and 30 women who live with long-term illness. FINDINGS: A recurring conversational thread has been women's experiences of fatigue when living with long-term illness. Although fatigue has been reported to be a major obstacle to maintaining usual daily activities and quality of life, few studies have explored this common symptom from the perspective of people themselves. Common themes found in the experience of fatigue as described by women are the meaning of fatigue, awareness as self-care, fatigue as invisible to others, seeking medical validation and accountability for self-care. CONCLUSION: It is vital for healthcare workers to give opportunities for women to talk about fatigue, validate their experiences and provide support with self-care. Healthcare workers are encouraged to challenge their own meanings and expectations surrounding a person's report of fatigue so that opportunities for therapeutic intervention can be facilitated. PMID- 16268842 TI - Introducing older people to the theory of gerotranscendence. AB - AIM: This paper reports a study whose aims were to introduce the theory of gerotranscendence to a group of older people; to give participants in the group an opportunity to discuss their ageing process; to study how participants described their ageing in relation to the theory; and to gather participants' opinions about discussing their ageing in a group. BACKGROUND: The theory of gerotranscendence states that human development is a process extending into old age. Guidelines had previously been derived for its practical use in the care of older people, aiming to promote their development towards gerotranscendence. METHODS: A qualitative descriptive approach was taken, and older people were invited to participate in group sessions at a day centre. At the sessions, participants discussed their ageing, and a video presentation about the theory of gerotranscendence was shown at one of the sessions. They were encouraged to discuss the description of the ageing process presented in the video and to link this to their own experiences of growing old. The discussion in each session was tape-recorded and transcribed verbatim. Data were analysed and categorized using qualitative methods. The data were collected in 2002. FINDINGS: All women had an experience of ageing that was in some way in line with the theory's description, and they more or less agreed that this description of ageing was in accordance with their own ageing. They considered that it was interesting and fruitful to discuss ageing in a group. They felt that introduction of the view of ageing offered by the theory of gerotranscendence was beneficial because it gave them a more positive view of ageing which also allowed them to be as they were. CONCLUSION: It is possible to arrange this type of group activity for older people, resulting in possibilities to use aspects of the theory of gerotranscendence as an intervention in gerontological nursing. PMID- 16268843 TI - Evaluation of an action research project in ophthalmic nursing practice. AB - AIM: This paper reports the evaluation phase of an action research project that promoted face-down posturing of patients following vitreo-retinal surgery for macular hole to enhance patient outcomes. The evaluation phase identified areas of practice needing further development from the perspectives of those involved with the care of patients. BACKGROUND: To achieve best results following surgical repair of macular hole, patients are required to posture face down for several weeks. As a consequence, patients complain of severe back and neck ache and find it difficult to persist with the posturing. Work to advance nursing practice as surgical developments occur has relevance beyond ophthalmology and the particular context of this project. METHOD: The first three phases of this action research- problem identification, planning and action--have been reported in another paper. Throughout the project an action research group comprising of representatives of key stakeholders were actively involved in researching and changing practice. During the evaluation phase, a qualitative methodology was chosen. Interviews with 17 members of staff from the inpatient area were carried out to elicit their perspectives on the posturing of patients. Qualitative interviews were selected to facilitate comparison with interview data from Phase 1. Data analysis ran concurrently with data collection, so that one could inform the other. FINDINGS: Overall, nurses and healthcare support workers felt that patients were more agreeable to posturing and after surgery began to posture more quickly. Communication was still an issue in some instances, and patients having urgent as opposed to planned surgery were found to be more difficult to prepare and the psychological care of patients still posed problems for nursing staff. CONCLUSIONS: The evaluation suggests that improvements in the care of this group of patients have occurred. A 10-point plan to promote face-down posturing has been developed which will be of use to practitioners in other settings. Some aspects of practice remain less well-understood, for example, the psychological care of patients. PMID- 16268844 TI - Preparing a large data set for analysis: using the minimum data set to study perineal dermatitis. AB - AIM: The aim of this paper is to present a practical example of preparing a large set of Minimum Data Set records for analysis, operationalizing Minimum Data Set items that defined risk factors for perineal dermatitis, our outcome variable. BACKGROUND: Research with nursing home elders remains a vital need as 'baby boomers' age. Conducting research in nursing homes is a daunting task. The Minimum Data Set is a standardized instrument used to assess many aspects of a nursing home resident's functional capability. United States Federal Regulations require a Minimum Data Set assessment of all nursing home residents. These large data would be a useful resource for research studies, but need to be extensively refined for use in most statistical analyses. Although fairly comprehensive, the Minimum Data Set does not provide direct measures of all clinical outcomes and variables of interest. METHOD: Perineal dermatitis is not directly measured in the Minimum Data Set. Additional information from prescribers' (physician and nurse) orders was used to identify cases of perineal dermatitis. The following steps were followed to produce Minimum Data Set records appropriate for analysis: (1) identification of a subset of Minimum Data Set records specific to the research, (2) identification of perineal dermatitis cases from the prescribers' orders, (3) merging of the perineal dermatitis cases with the Minimum Data Set data set, (4) identification of Minimum Data Set items used to operationalize the variables in our model of perineal dermatitis, (5) determination of the appropriate way to aggregate individual Minimum Data Set items into composite measures of the variables, (6) refinement of these composites using item analysis and (7) assessment of the distribution of the composite variables and need for transformations to use in statistical analysis. RESULTS: Cases of perineal dermatitis were successfully identified and composites were created that operationalized a model of perineal dermatitis. CONCLUSION: Following these steps resulted in a data set where data analysis could be pursued with confidence. Incorporating other sources of data, such as prescribers' orders, extends the usefulness of the Minimum Data Set for research use. PMID- 16268845 TI - Oral hygiene care for residents with dementia: a literature review. AB - AIM: This paper presents a literature review of oral hygiene care for adults with dementia in residential aged care facilities, including evidence for: (1) prevalence, incidence, experiences and increments of oral diseases; (2) use of assessment tools to evaluate residents' oral health; (3) preventive oral hygiene care strategies; and (4) provision of dental treatment. BACKGROUND: The impact of dementia on residential care is ever-increasing and regular oral hygiene care provision is challenging for cognitively impaired residents. Although an abundance of oral hygiene care recommendations for older people have been published, the supporting evidence has not been clearly delineated. METHODS: A review was conducted of English language publications (1980-2002), using a two step approach (keyword electronic database search, supplemented with secondary search of cited references). All 306 selected articles were critically reviewed and systematically categorized. RESULTS: Evidence confirmed clinicians' observations of poor oral health in older residents with dementia. Possible risk factors identified were: salivary dysfunction, polypharmacy, medical conditions, swallowing and dietary problems, functional dependence, oral hygiene care assistance and poor use of dental care. One comprehensive, reliable and validated oral assessment screening tool for residents with dementia had been published. Expert opinion indicated that oral assessment screening by staff and a dentist would be ideal at admission and regularly thereafter. Clinicians and researchers suggested that oral hygiene care strategies were effective in preventing oral diseases and appropriate for residents with dementia. CONCLUSION: These literature review findings supported the use of oral assessment screening tools by staff and efficacious preventive oral hygiene care strategies/products for adults with dementia in residential care facilities. Further research with this population is needed to develop and validate oral assessment tools and staff education programmes, trial preventive oral hygiene care strategies/products and trial dementia-focused behaviour management and communication strategies. PMID- 16268846 TI - Effects of timed voiding for the management of urinary incontinence in adults: systematic review. AB - AIM: The aim of this paper is to present a systematic review assessing the effectiveness of timed voiding for the management of urinary incontinence in adults. BACKGROUND: Despite the widespread use of systematic voiding programmes, their effectiveness is unclear, and the evidence for timed voiding has not been subject to rigorous and systematic evaluation. The impact on psychosocial factors and cost is also untested. The physiological basis for timed voiding is also poorly established. METHODS: The systematic review incorporated the methodology of the Cochrane Collaboration. All randomized or quasi-randomized controlled trials that addressed timed voiding for the management of urinary incontinence in adults were searched, appraised, analysed and summarized. The date of the latest search was 2002. Data were extracted independently and appraised according to the level of concealment of random allocation prior to formal entry; few and identifiable withdrawals and dropouts and an analysis based on an intention to treat. The relative risk for dichotomous data was calculated with 95% confidence intervals. Where data were insufficient to support quantitative analysis, a narrative overview was undertaken. RESULTS: Two trials of timed voiding met the inclusion criteria. In both, timed voiding was combined with other strategies. Participants were predominantly cognitively and physically impaired older women who resided in nursing home settings. Within-group improvements for the intervention groups were reported for both trials. One trial additionally reported a statistically significant reduction in night-time incontinence for the intervention group. The quality of the trials was modest and interpretation was limited by the potential for bias associated with inadequate concealment, missing data and no analysis by intention to treat. CONCLUSION: Terms used to describe voiding programmes that involve a fixed interval of voiding are variable. No conclusions can be drawn at this point about the effectiveness of timed voiding for the management of urinary incontinence in adults. PMID- 16268847 TI - Barriers to evidence-based practice in primary care nursing--why viewing decision making as context is helpful. AB - AIM: This paper reports a study examining the barriers associated with research knowledge transfer amongst primary care nurses in the context of clinical decision-making. BACKGROUND: The research literature on barriers to nurses' use of research knowledge is characterized by studies that rely primarily on self report data, making them prone to reporting biases. Studies of the barriers to evidence-based practice often fail to examine information use and behaviour in the context of clinical decision-making. METHODS: A multi-site, mixed method, case study was carried out in 2001. Data were collected in three primary care organizations by means of interviews with 82 primary care nurses, 270 hours of non-participant observation and 122 Q-sorts. Nurses were selected using a published theoretical sampling frame. Between-methods triangulation was employed and data analysed according to the principles of constant comparison. Multiple linear regression was used to explore relationships between a number of independent demographic variables (such as length of clinical experience) and the dependent variable of nurses' perspectives on the barriers to their use of research knowledge. RESULTS: Three perspectives on barriers to research information use emerged: the need to bridge the skills and knowledge gap for successful knowledge transfer; information formats need to maximize limited opportunities for consumption; and limited access in the context of limited time for decision-making and information consumption. Demographic variables largely failed to predict allegiance to any of the perspectives identified. CONCLUSIONS: Researchers should consider using decision-making as a contextual backdrop for exploring information use and behaviour, avoid relying solely on self-reported behaviour as data, and use a variety of research methods to provide a richer picture of information-related behaviour. Practice developers need to recognize that understanding the decisions to which research knowledge is to be applied should be a characteristic of any strategy to increase research uptake by nurses. PMID- 16268848 TI - Daytime sleepiness, sleep habits and occupational accidents among hospital nurses. AB - AIM: This paper reports a study to determine the prevalence of excessive daytime sleepiness and sleep habits among hospital nurses and to analyse associations between excessive daytime sleepiness and different types of medical error. BACKGROUND: It has been reported that sleep disorders, and the tiredness and sleepiness brought about by sleep disorders may be associated with occupational accidents. However, to our knowledge, there has so far been no report on associations between sleep disorders, excessive daytime sleepiness in particular, and occupational accidents among hospital nurses. METHODS: The study was a cross sectional study targeting 4407 nurses working in eight large general hospitals in Japan. An anonymous self-administered questionnaire was used to investigate their sleep patterns and experience of occupational accidents. The data were collected in 2003. RESULTS: The prevalence of excessive daytime sleepiness among hospital nurses in the present study was 26.0%. A statistically significant relationship was observed between having or not having occupational accidents during the past 12 months and excessive daytime sleepiness. Multiple logistic regression analyses on factors leading to occupational accidents during the past 12 months showed statistically significant associations between (1) drug administration errors and (2) shift work and age, between (1) incorrect operation of medical equipment and (2) excessive daytime sleepiness and age, and between needlestick injuries and age. CONCLUSIONS: Excessive daytime sleepiness is an important occupational health issue in hospital nurses. It is possible that occupational policies and health promotion measures, such as a provision of sleep hygiene advice and social support at worksites, would be effective in preventing occupational accidents among hospital nurses. PMID- 16268849 TI - Trust and managerialism: exploring discourses of care. AB - AIM: This paper reports a study that explored the relationship between trust and managerialism through an analysis of professional discourse. BACKGROUND: Managerialism is a distinct set of discourses and practices related to managerial effectiveness, flexibility and consumer responsiveness that have come to characterize debates over the provision of health and welfare services across the developed world. At the same time, trust has attracted increasing academic and political interest. Managerial discourses are critical of healthcare professions and the way they operate. Professional opinions are challenged as representing the interests of professions rather than service users; as a consequence trust is contested. However, where practitioners are both professionals and managers, the boundaries between these discourses become blurred. Moreover, paradoxical development occurs where increasing autonomy for practitioners is accompanied by a strengthening of managerial controls over their activity. METHOD: Discourse analysis was used to explore the text from two genres, academic literature and interviews (n = 17), in the context of community residential services for people with learning disabilities. The study was conducted in 2001. FINDINGS: Two broad themes were identified, each with a number of sub-themes. The first focuses on the relationship between managerialism and trust located around the management of expectations. The second, 'the politics of care', explores the way professional and managerial discourse articulate to produce complementary and contradictory positions. CONCLUSION: The colonization of professional activity by managerial discourse has produced a context where professional activity is defined by a series of managerial imperatives; trust, which was once the product of intimate social activity is now shaped through techniques based on distrust such as audits and quality monitoring. Nevertheless, the persistence of tensions between trust and managerialism suggest an ongoing struggle for professional autonomy in the face of increasing managerial controls. PMID- 16268852 TI - Packaging: a grounded theory of how to report physiological deterioration effectively. AB - AIM: The aim of this paper is to present a study of how ward-based staff use vital signs and the Early Warning Score to package physiological deterioration effectively to ensure successful referral to doctors. BACKGROUND: The literature tends to emphasize the identification of premonitory signs in predicting physiological deterioration. However, these signs lack sensitivity and specificity, and there is evidence that nurses rely on subjective and subtle indicators. The Early Warning Score was developed for the early detection of deterioration and has been widely implemented, with various modifications. METHOD: The data reported here form part of a larger study investigating the practical problems faced by general ward staff in detecting physiological deterioration. During 2002, interviews and observations were carried out using a grounded theory approach, and a total of 44 participants were interviewed (30 nurses, 7 doctors and 7 health care support workers). FINDINGS: Participants reported that quantifiable evidence is the most effective means of referring patients to doctors, and the Early Warning Score achieves this by improving communication between professionals. Rather than reporting changes in individual vital signs, the Early Warning Score effectively packages them together, resulting in a much more convincing referral. It gives nurses a precise, concise and unambiguous means of communicating deterioration, and confidence in using medical language. Thus, nurses are empowered and doctors can focus quickly on identified problems. CONCLUSION: The Early Warning Score leads to successful referral of patients by providing an agreed framework for assessment, increasing confidence in the use of medical language and empowering nurses. It is essential that nurses and nursing students are supported in its use and in developing confidence in using medical language by continued emphasis on physiology and pathophysiology in the nursing curriculum. PMID- 16268853 TI - Thermal management for premature births. AB - AIM: This paper reports an audit of the effect on admission temperatures of using occlusive polyethylene wrap applied immediately after the birth of extremely premature infants. BACKGROUND: Use of occlusive polyethylene wrap during the early postnatal management of the premature infant reduces evaporative and convective heat loss. METHOD: Retrospective pre-intervention audit was carried out, followed by the introduction of occlusive polyethylene wrap for thermal management during resuscitation and early stabilization. Prospective post intervention audit was then performed. The pre-intervention (control) group infants were immediately dried with prewarmed towels and resuscitated under radiant heat. Infants in the intervention group were managed under radiant heat, were not dried but were immediately enclosed in an occlusive polyethylene wrap. RESULTS: The demographic characteristics of the two groups were comparable. Use of occlusive polyethylene wrap resulted in higher admission temperatures for infants less than 27 weeks gestation (z=108.50, P<0.01). There was no statistically significant improvement in admission temperatures for 27-29 week infants. The rate of hypothermia on admission (<35.6 degrees C per axilla) was lower in the intervention group (chi(2)=5.12, d.f.=1, P=0.02), but more infants recorded temperatures exceeding 37.2 degrees C during the first 12 hours (chi(2)=23.45, d.f.=1, P<0.01). There were no other adverse effects noted. CONCLUSION: Use of occlusive polyethylene wrap improved admission temperatures for infants less than 27 weeks gestation. This intervention is easy to implement and does not interfere with resuscitation. However, removal of the wrap should be considered following admission to a closed care system in the neonatal intensive care unit because, in the intervention group, hyperthermia in the first 12 hours was a potential side effect. PMID- 16268854 TI - Determinants of parental behaviour when children undergo day-care surgery. AB - AIMS: This paper reports an investigation of the nature and strength of parental beliefs about their participation in their children's day-care surgery as well as demonstrated parental behaviours during the immediate post-recovery period. BACKGROUND: Parents want to participate in their children's hospital care but their behaviour does not reflect their intention. Parents' beliefs about their role in the hospital may be a factor influencing the way they help their children at the course of a day-care surgery. RESEARCH METHODS: A descriptive correlative design was used. French and English speaking parents of 3-12-year old children were asked to complete a self-administered questionnaire which comprised 42 items measuring the intensity of attitudes, norms and sense of control and their influence in predicting their intention to participate in hospital care. Parents' behaviours during the first hour following the child's return from the recovery room were video-recorded. RESULTS: A total of 220 parents participated in the study. Parental beliefs about participation in care were quite strong in terms of subjective norms, sense of control and attitude, predicting 64% of their intention to participate. Parents demonstrated mostly attitude type behaviours, while the level of demonstrated helping behaviours was somewhat low. CONCLUSIONS: Parents can be valuable partners in ambulatory care settings. Clinicians and health care managers should aim at establishing a true partnership with parents, in order to improve the quality of care given to children and their family in hospital and at home. PMID- 16268855 TI - Responses to advanced cancer: Chinese-Australians. AB - AIM: This paper describes a study identifying the impact of key aspects of Chinese culture on the responses of mid-aged Chinese-Australians to their advanced cancer in order to make recommendations about their care within the health system. BACKGROUND: Studies conducted in the 1960s and 1970s focused on understanding people's psychological responses to their experiences of terminal illness, but the issue of culture was not addressed. In recent years, a few studies have been conducted with Chinese-Australians, but were limited to issues related to their information needs and the disclosure of a cancer diagnosis. There is a lack of understanding of the impact of Chinese culture on the experiences of these patients. METHOD: A grounded theory approach was used to generate a substantive theory to explain how mid-aged Chinese-Australians respond to advancing cancer. Eleven participants were recruited and data were collected from face-to-face interviews, telephone contacts, observation and researcher field notes. Data generation occurred between 1997 and 1999. FINDINGS: Four modes of response to advanced cancer were identified: acute crisis, combat, despondency and waiting for death. This paper deals particularly with the combat mode which incorporated five culturally specific strategies used by participants in their struggle against advanced cancer. These were traditional Chinese medicine, traditional Chinese beliefs on the use of food for health maintenance, qi gong (a form of exercise), feng shui (which involves paying attention to spatial organization) and the worship of ancestors and gods. Deeply entrenched within these responses is the influence of Chinese culture, rooted in the beliefs and practices of traditional Chinese medicine and the philosophy of harmony and balance of yin and yang and qi. CONCLUSION: Health care professionals need to be aware of the cultural practices and beliefs of the different ethnic groups for whom they care, and of the importance of accommodation to and negotiation about these cultural practices. PMID- 16268856 TI - Hope seen through the eyes of 10 Australian young people. AB - AIM: The aim of this paper is to report the findings of a phenomenological study that explored hope in 10 young people in Australia. BACKGROUND: Evidence suggests many Australian young people are in crisis. Examination of key reports that detail the incidence of suicide, early drug-taking behaviours, homelessness, self harm behaviours, joblessness, depressive disorders, crime statistics and alcohol abuse suggest that many of today's young people have lost resilience as well as vital connections to their community. METHOD: Two methods were employed to encourage the participants to reflect on their experiences of hope - what it is and what it meant to them. The first was to supply participants with a disposable colour film camera and ask them to take pictures that, in their view, showed hope. The second was participation in an in-depth interview that was prompted in part, by their photographs. Interview audiotapes were transcribed verbatim and analysis of the text used the Turner method. The data were collected in 2002. FINDINGS: Four horizons of hope were revealed: at-one-with; a driving force; having choices; and connecting and being connected. These horizons are discussed, showing how, or if, the literature treats these dimensions of hope. Perspectives are offered on how they might be considered by nurses who are charged with caring for today's young people. CONCLUSION: Registered Nurses who work with young people must understand the phenomenon of hope from their unique perspective before they can offer appropriate hope-facilitating strategies. PMID- 16268857 TI - Put hope to work: a commentary. PMID- 16268858 TI - Culture and ageing: reflections on the arts and nursing. AB - AIM; In this paper, we focus on ageing as an area in which nursing, society and the humanities can be profitably conjoined. We illustrate our argument with three case studies of ageing: in painting, opera and ballet. BACKGROUND: There has been a recent spectacular increase in papers devoted to the relatively new field of the medical humanities. We argue for a similar renaissance in thinking about the connections between the arts and nursing. DISCUSSION: First, we consider the paintings of J.M.W. Turner (1775-1851) as examples of loss, ageing and death. Second, we draw upon Leos Janacek's opera 'The Makropulos Case' (1926) as a focus for debate about human mortality. Third, we review some ethnographic research on the balletic body as an example of cultures of youthful ageing. CONCLUSION: A focus on the embodiment of vulnerability is a productive catalyst for research on the intimate connections between self and society, biology and culture, and reason and emotion. Such a research agenda would be the hallmark of a holistic approach to the arts and nursing. PMID- 16268859 TI - Agitation in dementia: concept clarification. AB - AIM: The aim of this paper is to clarify the concept of agitation in dementia through analysing definitions, critical attributes, components, boundaries, antecedents and consequences of agitation. BACKGROUND: The concept of agitation is not well defined. In addition, there exists much confusion about the characteristics and boundaries of agitation, as well as the distinction between agitation and related concepts. Recently developed theoretical models for agitation in dementia require new interpretation and conceptualization of agitation. METHODS: Morse's method of critical appraisal of the literature was used. In addition, some parts of Rodgers' evolutionary method were employed. Data were selected using six electronic databases and the key words 'agitation', 'agitated', 'dementia', 'demented' and 'Alzheimer'. The analysis included 86 empirical or theoretical papers and one book. RESULTS: A transition from the observer's perspective to the patient's perspective in the interpretation of agitation was found. Five critical attributes of agitation in dementia were identified: excessive, inappropriate, repetitive, non-specific and observable. Patient factors, interpersonal factors, environmental factors and restraint were identified as precipitating antecedents. Mediating antecedents included discomfort, unmet need and misinterpretation. Consequences of agitation were identified at the levels of patient, caregiver and others. CONCLUSIONS: This transition in perspectives has important implications as it can change health providers' attitudes and responses to agitation and lead to patient-focused and individualized care. Researchers and clinicians are encouraged to avoid labelling agitated behaviour as 'disturbing behaviour'. PMID- 16268860 TI - The ethics of research using electronic mail discussion groups. AB - AIM: The aim of this paper is to identify and discuss the ethical considerations that have confronted and challenged the research team when researchers facilitate conversations using private electronic mail discussion lists. BACKGROUND: The use of electronic mail group conversations, as a collaborative data generation method, remains underdeveloped in nursing. Ethical challenges associated with this approach to data generation have only begun to be considered. As receipt of ethics approval for a study titled; 'Describing transition with people who live with chronic illness' we have been challenged by many ethical dilemmas, hence we believe it is timely to share the issues that have confronted the research team. These discussions are essential so we can understand the possibilities for research interaction, communication, and collaboration made possible by advanced information technologies. DISCUSSION: Our experiences in this study have increased our awareness for ongoing ethical discussions about privacy, confidentiality, consent, accountability and openness underpinning research with human participants when generating data using an electronic mail discussion group. We describe how we work at upholding these ethical principles focusing on informed consent, participant confidentiality and privacy, the participants as threats to themselves and one another, public-private confusion, employees with access, hackers and threats from the researchers. CONCLUSION: A variety of complex issues arise during cyberspace research that can make the application of traditional ethical standards troublesome. Communication in cyberspace alters the temporal, spatial and sensory components of human interaction, thereby challenging traditional ethical definitions and calling to question some basic assumptions about identity and ones right to keep aspects of it confidential. Nurse researchers are bound by human research ethics protocols; however, the nature of research by electronic mail generates moral issues as well as ethical concerns. Vigilance by researchers is required to ensure that data are viewed within the scope of the enabling ethics approval. PMID- 16268861 TI - The integrative review: updated methodology. AB - AIM: The aim of this paper is to distinguish the integrative review method from other review methods and to propose methodological strategies specific to the integrative review method to enhance the rigour of the process. BACKGROUND: Recent evidence-based practice initiatives have increased the need for and the production of all types of reviews of the literature (integrative reviews, systematic reviews, meta-analyses, and qualitative reviews). The integrative review method is the only approach that allows for the combination of diverse methodologies (for example, experimental and non-experimental research), and has the potential to play a greater role in evidence-based practice for nursing. With respect to the integrative review method, strategies to enhance data collection and extraction have been developed; however, methods of analysis, synthesis, and conclusion drawing remain poorly formulated. DISCUSSION: A modified framework for research reviews is presented to address issues specific to the integrative review method. Issues related to specifying the review purpose, searching the literature, evaluating data from primary sources, analysing data, and presenting the results are discussed. Data analysis methods of qualitative research are proposed as strategies that enhance the rigour of combining diverse methodologies as well as empirical and theoretical sources in an integrative review. CONCLUSION: An updated integrative review method has the potential to allow for diverse primary research methods to become a greater part of evidence-based practice initiatives. PMID- 16268862 TI - The value of integrating interpretive research approaches in the exposition of health care context. AB - AIM: This paper discusses the use of a nested set of methodologies (dramaturgy, ethno-methodology and ethnography) to characterize and interpret the settings, practices and interactions inherent in the health care environment. The aim is to explain how a set of methodologies can help make sense of research data in the clinical setting. BACKGROUND: Despite the recognition of the importance of the context of care there has been limited debate about the use and value of research methods and methodologies and how they can be best applied to the health care context. DISCUSSION: Using dramaturgy the physical and social scene can clearly be established, to enable insight into 'how the scene is contrived'. The ethno methodological approach assists in the examination of taken-for-granted assumptions inherent in the interactions between individuals in the 'scene', and the underlying 'shared' knowledge within interactions. 'Shared knowledge' identifies knowledge as a medium for communication. The use of ethnography ensures that social and cultural symbols, which are an integral component of how individuals collectively attribute meaning to places and events, become a significant part in the interpretation of interactions. CONCLUSION: The combination of these methods is advantageous in assisting qualitative researchers in the health care environment to 'make sense' of their complex field notes. PMID- 16268863 TI - Therapeutic working relationships with people with schizophrenia: literature review. AB - AIM: The aim of this paper is to review the evidence for the necessity and sufficiency of therapeutic relationships when working with people with enduring mental health problems, such as schizophrenia. BACKGROUND: The value of therapeutic relationships in mental health nursing has been the subject of some debate within the profession. This debate has centred on the spectrum of beliefs about therapeutic relationships, ranging from the position that the relationship is both necessary and sufficient to enable change, to more technical approaches, to therapeutic intervention which de-emphasises the influence of the relationship. METHODS: Searches for published material in English between 1986 and 2003 were carried out using the following databases: Cumulative Index of Nursing and Allied Health Literature; MEDLINE; Applied Social Sciences Index and Abstracts; Sociological abstracts; and social service abstracts. The search terms were: therapeutic alliance; therapeutic relationship; working alliance; and nurse patient relationships. Papers chosen for inclusion in the review were those with a research focus on the elements and potential benefits/costs of therapeutic relationships in nursing. RESULTS: People who experience a relationship as being therapeutic appear to have better outcomes. A consistent finding of a number of meta-analyses is that therapeutic relationships characterized by facilitative and positive interpersonal relationships with the helper have in-built benefits, and that this is an important element of advanced techniques. In order for cognitive behavioural therapy to be successful, people need to feel understood and involved in the therapeutic relationship. CONCLUSION: Therapeutic relationships are necessary but not sufficient to enable change when working with people with schizophrenia. PMID- 16268864 TI - A response to T. Koch, P. Jenkin & D. Kralik (2004) Chronic illness self management: locating the 'self'. PMID- 16268869 TI - Review of the potential photo-cocarcinogenicity of topical calcineurin inhibitors: position statement of the European Dermatology Forum. AB - Topical Calcineurin Inhibitors (TCIs) used for the treatment of atopic eczema modify the immune regulatory function of the skin and may have the potential to enhance immunosuppressive ultraviolet (UV) effects. Current recommendations on UV protection in eczema patients treated with PCIs are inconsistent and have given rise to uncertainty and anxiety in patients. Therefore, the European Dermatology Forum (EDF) developed a position statement which reviews critically the available data with regard to the problem, especially analysing and commenting the limitations of rodent models for the human situation. There is no conclusive evidence from rodent trials to indicate that long-term application of TCIs is photococarcinogenic. There is a need for further studies to investigate the validity of mouse models as well as long-term cohort studies in patients using TCIs. Available data suggest that long-term application of TCIs is safe, that there is no evidence of increased skin cancer risk and that it is ethical to treat patients with TCIs when indicated. PMID- 16268870 TI - The clinical benefit of moisturizers. AB - Moisturizing creams marketed to consumers often contain trendy ingredients and are accompanied by exciting names and attractive claims. Moisturizers are also an important part of the dermatologist's armamentarium to treat dry skin conditions and maintain healthy skin. The products can be regarded as cosmetics, but may also be regulated as medicinal products if they are marketed against dry skin diseases, such as atopic dermatitis and ichthyosis. When moisturizers are used on the so-called dry skin, many distinct disorders that manifest themselves with the generally recognized symptoms of dryness are treated. Dryness is not a single entity, but is characterized by differences in chemistry and morphology in the epidermis depending on the internal and external stressors of the skin. Patients and the society expect dermatologists and pharmacists to be able to recommend treatment for various dry skin conditions upon evidence-based medicine. LEARNING OBJECTIVE: Upon completing this paper, the reader should be aware of different types of moisturizers and their major constituents. Furthermore, s/he will know more about the relief of dryness symptoms and the functional changes of the skin induced by moisturizers. PMID- 16268871 TI - An effective surgical treatment for nail thickening in Darier's disease. AB - BACKGROUND: Nail thickening of idiopathic or hereditary aetiology originating in the matrix is difficult to treat. OBJECTIVE; The purpose is to describe an easy surgical technique to reduce the thickness of the nail. METHODS AND RESULTS: The result is achieved by shortening the matrix with an excellent cosmetic appearance in the newly formed nail plate. CONCLUSION: Nail thickening in Darier's disease originating in the matrix, and other similar hereditary or idiopathic conditions, may benefit from this surgical approach. PMID- 16268872 TI - Interaction of legislation and prevalence of sexually transmitted diseases with focus on syphilis and gonorrhoea in the Czech Republic - review to year 2003. AB - INTRODUCTION: A large increase in syphilis prevalence was noted in the Czech Republic during the last decade. It was associated with prostitution in cities and especially in areas bordering more economically developed countries. Foreigners account for more than 50% of this increase. Absence of legislative regulations contributes to this situation. OBJECTIVE: To demonstrate the necessity of setting sexually transmitted disease (STD) fighting priority in the Czech Republic. METHODS: The authors review the development of legislation related to STDs in the context of geopolitical changes and the epidemiological situation in the Czech Republic by relating epidemiological, geopolitical and economic data. RESULTS: Venereal-diseases-related regulatory codes have existed in the Czech Republic for years; however, they became fragmented in several regulations and often do not have the status of law. It is a consequence of the transformation of the entire state health care delivery system to a private system. The treatment of foreigners, refugees and uninsured people is a major problem. CONCLUSION: It is necessary to focus on special populations, such as homeless people, immigrants and prostitutes. It is desirable to strengthen international cooperation in combating STD as well as prostitution. Relevant legislation should be introduced to correlate the existing regulations and bring them up to date. PMID- 16268873 TI - Women dermatologists and the Spanish Academy of Dermatology and Venereology (SADV). AB - BASIS: The Spanish Academy of Dermatology and Venereology (SADV) consists of a large majority of Spanish dermatologists. Their track record and history can be considered as parallel to that of female dermatologists. The aim of this study is to determine the role of women in the SADV throughout its history, both in terms of their involvement and their positions of responsibility. MATERIALS AND METHODS: To obtain data, we consulted the SADV archives from the year of its foundation until 2001. We also consulted other Spanish medical societies, receiving responses from three of them. RESULTS: Until 1964, no woman had ever become a member of the SADV. Since then, the percentage began to increase slowly (5.5% in 1975). Over the last decade, their involvement has increased rapidly, reaching 41.34% in 2001. During this time, only 11 women have held positions on the Board of Directors, and in all cases in nonpresidential positions. The percentage of women in paediatrics, ophthalmology and pathologic anatomy is similar. CONCLUSIONS: During this time, the number of female dermatologists in the SADV has gradually increased. This increase is, nonetheless, not uniform, with the greatest growth being seen during the last decade. Women in positions of responsibility have been scarce and, to date, there has been no female national president. These results coincide with statistics revealed in other areas of the medical society. PMID- 16268874 TI - Frontal fibrosing alopecia: a survey in 16 patients. AB - BACKGROUND: Postmenopausal frontal fibrosing alopecia (PFFA) was described by Kossard et al. as a progressive recession of the frontal hairline affecting particularly postmenopausal women. Further cases of PFFA have been reported to date, all of them considering it as a variant of lichen planopilaris on the basis of its clinical, histological and immunohistochemical features. OBJECTIVE: To describe clinical features, and response to treatment of 16 cases of frontal fibrosing alopecia diagnosed at our department in the last 6 years. METHODS: In addition to clinical data, biopsies and laboratory tests (antinuclear antibodies, sex hormones, thyroid hormones) were performed in order to rule out other causes of scarring alopecia. Patients were treated with intralesional corticosteroids, finasteride, and minoxidil, depending on the stage of the disease and association to androgenetic alopecia. RESULTS: All patients presented progressive alopecia localized to the frontal and temporal hairlines. Eight patients (50%) had loss of eyebrows, and six patients (37.5%) had axillar alopecia. Ages ranged from 45 to 79. Three of these women were premenopausal. Androgenetic alopecia was evident in seven patients (43.8%). All patients biopsied showed perifollicular lymphocitic infiltrate with lamelar fibrosis limited to the upper portions of the follicle. The progression of the condition stopped in most patients after a variable period on treatment. When treatment was abandoned the alopecia progressed to 'clown alopecia' appearance. DISCUSSION: Cases of Kossard's type scarring alopecia affecting premenopausal women made us consider that this condition is not exclusive of postmenopausal women. Differential diagnosis should take into account conditions like female androgenetic alopecia, fibrosing alopecia in a pattern distribution, alopecia areata, and chronic lupus erythematosus. Except for the pattern of alopecia, lichen planopilaris and frontal fibrosing alopecia are indistinguishable, thus the latter is included as a variant of lichen planopilaris. Although the disease tends to spontaneous stabilization, intralesional and topical corticosteroids, and anti-androgens may stop the progression of the disease and improve the female androgenetic alopecia that usually is associated to FFA. PMID- 16268875 TI - Assessment of the immune system in 55 Iranian patients with vitiligo. AB - BACKGROUND: Vitiligo is an acquired idiopathic hypomelanotic disorder characterized by circumscribed depigmented macules resulting from the loss of cutaneous melanocytes. OBJECTIVE: In order to evaluate the immune system of Iranian patients with vitiligo, this study was accomplished. METHODS: Fifty-five Iranian patients with vitiligo and 60 healthy persons as control were investigated in this study. The laboratory techniques were included: antimelanocyte antibody (AMA) and antinuclear antibody (ANA) with indirect immunoflorescent test, C3 and C4 levels with single radial immunodiffusion (SRID), and rheumatoid factor (RF) with enzyme-linked immunosorbent assay (ELISA). RESULTS: AMA was positive in 17 patients (30.9%) and was negative in the entire control group (P < 0.0001). ANA was positive in 4 patients (7.3%), which was insignificantly higher than control group (1.7%). IgM-RF was positive in 6 patients (10.8%) while it was negative in the entire control group (P = 0.027). C3 and C4 values decreased in 14 patients (25.5%), which was significantly higher than control group (P < 0.001). CONCLUSION: The important role of the immune system in the pathogenesis of vitiligo could be suggested. In addition, the autoimmune hypothesis of vitiligo could be confirmed based on the results of this study. PMID- 16268876 TI - Effects of patient satisfaction with care on health-related quality of life: a prospective study. AB - OBJECTIVES: To evaluate the effect of patient satisfaction on health-related quality of life among dermatological outpatients, independently of patients' sociodemographic, and clinical characteristics. DESIGN: Prospective cohort study. Satisfaction was evaluated with a standardized questionnaire three days after the visit by telephone interview. Quality of life and psychiatric disorders were measured with validated instruments (Skindex-29 and Ghq-12) before the dermatological visit and after four weeks. At the four week interview also self reported medication adherence was assessed. SETTING: Outpatient clinics of a large dermatological hospital in Rome, Italy. PATIENTS: A consecutive sample of 1389 outpatients was approached, and 52% agreed to participate. Inclusion criteria were met by 424 patients, and 396 (93%) of them completed the study. MAIN OUTCOME MEASURES: Improvement in health related quality of life. RESULTS: At multivariate analysis satisfied patients have approximately twice the odds for an improved quality of life on the emotions scale (OR = 1.99; 95%CI 1.1 to 3.7; P = 0.03) and on the functioning scale (OR = 2.2; 95%CI 1.1 to 4.7; P = 0.03). Patients with psychiatric disorders at baseline were less likely to have an improved quality of life on the functioning scale (OR = 0.3; 95%CI 0.2 to 0.8; P = 0.01). CONCLUSIONS: This is the first prospective study showing that patient satisfaction and psychiatric disorders have a significant effect on quality of life improvement among dermatological patients, independently of patient characteristics. Particular attention should be devoted to improving physicians' interpersonal skills, the major component of patient satisfaction. PMID- 16268877 TI - Perifolliculitis capitis abscedens et suffodiens. AB - Perifolliculitis capitis abscedens et suffodiens (PCAS) is rare chronic, suppurative and inflammatory scalp disease. Its aetiology and pathogenesis is not completely understood. The treatment is usually difficult and often disappointing. We report a case of 29-year-old male who presented with tender, fluctuant nodules and abscesses, with draining pus and patchy alopecia on his scalp for 3 years. A skin biopsy from scalp lesions revealed features that are characteristic of perifolliculitis. Initially, the patient was treated with periodic incision and drainage of the scalp abscesses. The answer was very poor. When admitted to our department, isotretinoin was started at daily dose of 30 mg, because initially his cholesterol and triglyceride levels were mildly increased. When dose was reduced to 10 mg the levels of cholesterol and triglyceride remained normal. A response to treatment was excellent and rapid. The treatment of PCAS represents usually difficulties and frustration for both the patient and the physician. A long course of isotretinoin can be considered as one of the most effective treatment for PCAS. PMID- 16268879 TI - Bilateral blepharochalasis. AB - Blepharochalasis is a rare disorder characterized by recurrent painless periorbital oedema, which leads to atrophy of the periorbital skin. Pathomechanism of the disease is probably immunological with a nearly complete loss of elastic fibres. The authors describe a 17-year-old woman, who was followed for 7 years. IgA deposits were found in the periorbital tissues, which confirm the immunological background of the condition. Electron microscopy results show that not only elastic but also collagen fibres were affected. PMID- 16268880 TI - Ross syndrome, an entity included within the spectrum of partial disautonomic syndromes. AB - Ross syndrome is a degenerative peripheral nervous system disorder defined by the following triad: unilateral or bilateral segmental anhidrosis, hyporeflexia of deep tendon reflexes and Adie's tonic pupils. The most disturbing symptom is segmental compensatory hyperhidrosis. It has only occasionally been reported in the dermatological literature. We present a 35-year-old woman with chronic hepatitis C who developed the characteristic triad of Ross syndrome within 1 month. The patient was otherwise healthy except for an aneurysm of the left medium brain artery not responsible for the syndrome. PMID- 16268878 TI - Thysanoptera dermatitis. AB - Thysanoptera dermatitis is caused by the bite of small (1-2 mm) insects (generally thrips). Thrips usually feed on the juices of vegetables but if they reach human skin they can suck the epidermal lymph after biting. The cutaneous lesions formed are small pink and itchy papules localized mainly on the trunk and the arms. Diagnosis is only possible by demonstrating that the insect is present inside one of the lesions of the skin. Thysanoptera dermatitis is often misdiagnosed as mosquito bites. Although self-resolving in a few days, Thysanoptera dermatitis should be known not only for the cultural and scientific training of the dermatologist but also to add more detailed diagnostic information to the generic diagnosis of 'entomodermatosis' and to confirm the benign evolution of the bites. PMID- 16268881 TI - Recurrent epithelioid angiosarcoma of the scalp simulating melanoma. A 10-year follow-up. AB - We present a case of an intriguing mesenchymal neoplasm of the scalp that recurred several times over 10 years before a final diagnosis was possible. The case was sent for expert opinions to various international dermatopathological authorities and was, for a long time, unanimously interpreted as malignant melanoma. This diagnosis was supported by immunohistochemical examinations demonstrating S-100 positivity. Nevertheless, the clinical behaviour, as well as some histopathological features raised doubt regarding the diagnosis. Only after the last recurrence, followed by a repeat extensive immunohistochemical study, the diagnosis of epithelioid angiosarcoma was made. Histologically malignant melanoma can be highly misleading and in literature, reports of misinterpreted cases of melanoma are published. In contrast, tumours that can simulate melanoma are also not infrequent and it is essential to perform immunohistochemistry to confirm diagnosis and exclude a melanocytic lesion. PMID- 16268882 TI - Melanoma associated with subacute primitive fibrinolysis. AB - BACKGROUND: Surgery is the first treatment of a pigmented skin lesion when a melanoma is suspected. This excision is most of the time realized by the dermatologist as well as the second time surgery. Bleeding at the surgical site may have numerous aetiologies. Fibrinolysis is a rare but dramatic event. CASE REPORT: We report a case with delayed bleeding after excision of a melanoma. The patient was re-operated and a major haemorrhage followed so that a transfusion of blood and fresh frozen plasma was necessary. The biological investigations concluded to a subacute primitive fibrinolysis associated with the melanoma. The patient was first considered to have completely recovered when a blood-borne liver metastasis diffusion rapidly occurred. The patient deceased within a few months from a hepatic encephalopathy with hepatic metastases. DISCUSSION: We find only a few published cases of melanoma with fibrinolysis. Its association with other abnormalities, like a partial factor XIII deficiency, made these abnormalities even more difficult to identify. The relation between the subacute fibrinolysis, the melanoma and the liver infiltrative metastasis is difficult to establish. PMID- 16268883 TI - Parry-Romberg syndrome in association with anti-dsDNA antibodies: a case report. AB - Parry-Romberg syndrome (PRS) is a rare and puzzling disorder that is characterized by progressive hemifacial atrophy. It involves mainly some or all tissues of one side of the face. A case of 21-year-old Caucasian man with hemifacial atrophy in the right facial region is reported. Serological studies with anti-single-stranded DNA (anti-ssDNA), anti-double-stranded DNA (anti dsDNA), anticentromere (ACA) and antinuclear (ANA) antibodies were done. Anti dsDNA antibodies was found positive, but the others were negative. Rheumatoid factor (RF) was also negative. Since PRS is rare and its association with anti dsDNA antibodies was not reported before, this case appears to be the first report. PMID- 16268884 TI - Impetigo herpetiformis with hyperparathyroidism. AB - Impetigo herpetiformis, first described by Hebra, is a rare pustular disorder that primarily affects pregnant women and it is often complicated by an increased risk of spontaneous abortion. A commonly associated hypocalcemia often appears with hypoparathyroidism. Here we report a case of complicated impetigo herpetiformis without hypocalcemia paired with a compensatory hyperparathyroidism. PMID- 16268885 TI - Subcutaneous Rosai-Dorfman disease: is surgical excision justified? AB - Rosai-Dorfman disease in soft tissue without nodal disease has been recognized as a distinct clinicopathologic entity. It may represent a diagnostic challenge and the natural history and optimal treatment has not been well clarified. We investigated a patient in whom Rosai-Dorfman disease was confined to the subcutis of the abdominal wall and recurred after incomplete excision. Complete resolution was achieved by wide surgical excision with negative margins. Pathologic examination confirmed the diagnosis of extranodal Rosai-Dorfman disease. The patient is disease-free after 1 year of follow-up. Despite the possibility that spontaneous remission may occur, our results suggest that when anatomically feasible, complete excision can be a treatment option for persistence or recurrence of exclusively extranodal disease. Larger case series and longer follow-up are needed to assess the long-term efficacy in these patients. PMID- 16268886 TI - Naevus lipomatosus cutaneous superficialis on the nose. AB - Naevus lipomatosus cutaneous superficialis (NLCS) is an uncommon hamartomatous lesion with an exceptional presentation on the face. We report the case of an elderly patient who presented with a classic type of NLCS on the right nasal orifice. This patient was taking inhaled steroids for his allergic rhinitis during 35 years. PMID- 16268887 TI - 'Dalmatian dog'-like skin eruption (two cases of multifocal fixed drug eruption induced by mefenamic acid). AB - Mefenamic acid is a common widely prescribed drug with analgesic activity. Authors report two cases of multifocal fixed drug eruption induced by mefenamic acid. Cases were diagnosed on basis of clinical examination and histopathology of skin lesion. Only a few cases have been reported in the literature and these are the first two described in Greece. PMID- 16268888 TI - Epidermoid carcinoma and perforating necrobiosis lipoidica: a rare association. AB - We report the case of a 33-year-old patient who had had insulin-dependent diabetes mellitus (IDDM) since he was 11 months old, and who presented with major perforating necrobiosis lipoidica (PNL) complicated by a well-differentiated epidermoid carcinoma. PNL is a rare clinical form of NL, always associated with diabetes. Only seven cases have been reported to date in the literature, and to the best of our knowledge, an association of epidermoid carcinoma and PNL has never been described. The development of a tumoral transformation on a classical NL plaque has only been described 12 times. The presence of an epidermoid carcinoma on a weakened background with permanent ulceration suggests that early surgical excision of the tumour and of the NL followed by a skin graft might be the treatment of choice. Radiotherapy seems to be a poor therapeutic option. PMID- 16268889 TI - Ichthyosis follicularis, alopecia and photophobia (IFAP) syndrome treated with acitretin. AB - We describe a 3-year-old male patient with the ichthyosis follicularis, alopecia and photophobia (IFAP) syndrome, who developed cutaneous and ocular involvement in infancy. In addition, he had growth retardation and borderline intelligence; no other systemic involvement was found on detailed investigation. A moderate response to acitretin therapy (1 mg/kg) administered for 6 months was observed, with improvement in cutaneous features and corneal erosions and no change in alopecia or photophobia. PMID- 16268890 TI - Bullous haemorrhagic cellulitis caused by Enterobacter cloacae. PMID- 16268891 TI - Atrophia maculosa varioliformis cutis: a case with extrafacial involvement and familial facial lesions. PMID- 16268892 TI - Glutathione peroxidase (GSH-Px) enzyme levels of patients with acne vulgaris. PMID- 16268894 TI - Alopecia areata in Down syndrome: a clinical evaluation. PMID- 16268895 TI - Linear lichen planopilaris of the face. PMID- 16268893 TI - Elevated serum parathyroid hormone levels in a patient with annular pustular psoriasis. PMID- 16268896 TI - Cutis verticis gyrata secondary to acne scleroticans capitis. PMID- 16268897 TI - Mondor's disease probably due to herpes zoster. PMID- 16268898 TI - Soluble TNF-receptors, naive CD31+ T cells and B-1a cells in psoriatic patients. PMID- 16268899 TI - Particularities of clinic expression in a case of Bloch-Sulzberger syndrome. PMID- 16268900 TI - Follicular cysts and hyperkeratoses as first manifestation, and involvement of the central nervous system as late manifestation of mycosis fungoides. PMID- 16268902 TI - Childhood onset of psoriatic onycho-pachydermo-periostitis (POPP). PMID- 16268901 TI - Sequential treatment of angiosarcoma of the back with liposomal doxorubicin and radiotherapy. PMID- 16268903 TI - Anogenital warts in an infant. PMID- 16268904 TI - Kikuchi's disease with skin lesions in a patient with SLE. PMID- 16268907 TI - Update in screening of lung cancer. AB - In the USA, lung cancer is the leading cause of cancer death. Earlier studies of CXR and sputum cytology screening conducted in the 1970s showed no mortality benefit. Accordingly, mass screening for lung cancer was abandoned and is not currently recommended. Recently, interest in lung cancer screening has been revived due to various reports showing an advantage of low-dose CT over CXR in detecting smaller size tumours and at an earlier stage. Although these reports generated much enthusiasm for screening among clinicians and the general public, the effectiveness of low-dose CT in reducing lung cancer-specific mortality rates has not been demonstrated. Large-scale randomized controlled trials are currently in progress to determine the efficacy of CXR and low-dose CT screening. This review highlights the advantages and limitations of current modalities for lung cancer screening. The cases for and against screening with currently available modalities are examined. Additional new screening modalities are also discussed. PMID- 16268908 TI - The effects of erythromycin on the viability and the secretion of TNF-alpha and TGF-beta1 and expression of connexin43 by human pleural mesothelial cells. AB - OBJECTIVE: The mechanism by which erythromycin produces pleurodesis remains unknown. The purpose of this study was to investigate the effects of erythromycin on human pleural mesothelial cell (HPMC) viability, the secretion of tumour necrosis factor-alpha (TNF-alpha) and transforming growth factor-beta(1) (TGF beta(1)) and the level of expression of connexin43. METHODOLOGY: HPMC were incubated with different concentrations of erythromycin. The inhibitory effects of erythromycin on HPMC growth were measured using a tetrazolium-based colorimetric assay. The levels of TNF-alpha and TGF-beta(1) in supernatants were measured by ELISA and levels of connexin43 were assessed by Western blot. RESULTS: Erythromycin injured HPMC in a dose and time-dependent manner. The secretion of both TNF-alpha and TGF-beta(1) by HMPC increased significantly when they were incubated with 100 mg/L erythromycin for 3 or 5 days. The levels of connexin43 in HPMC decreased after incubation with 100 mg/L erythromycin and no relationship was observed between the levels and incubation time. CONCLUSIONS: Erythromycin injures HPMC in a dose- and time-dependent manner and results in the secretion of TNF-alpha and TGF-beta(1). This is one possible mechanism of pleurodesis with erythromycin. Furthermore, erythromycin decreased the levels of connexin43 in HPMC, which could possibly affect the response of HPMC to pleurodesis with erythromycin. PMID- 16268909 TI - Serum IgG and IgA antibodies to Chlamydia pneumoniae and severity of emphysema. AB - OBJECTIVE: Chronic Chlamydia pneumoniae infection has been identified serologically in patients with COPD. The aim of this study was to examine whether the severity of emphysema is related to elevated antibody titres against C. pneumoniae. METHODOLOGY: We measured antibody titres against C. pneumoniae using ELISA, and assessed the severity of emphysema by the percentage of low attenuation area (%LAA) using high resolution (HR) CT in patients with COPD and in non-smoking control subjects. RESULTS: The mean %LAA was 2.2% in non-smoking controls (n = 28) and 13.3% in COPD patients (n = 94). COPD patients with a high IgG antibody index to C. pneumoniae (> or =2.0, n = 42) had a significantly higher %LAA (16.8%) than those with a low IgG index (<2.0, n = 52) (10.6%, P = 0.01). In addition, COPD patients with a high IgA antibody index (> or =2.0, n = 46) had a significantly higher %LAA (15.9%) than those with a low IgA index (<2.0, n = 48) (10.9%, P = 0.048). COPD patients with a high IgA antibody index also had a significantly lower %DLco than that associated with a low IgA index (68.1% and 80.3%, respectively, P = 0.007). There were no significant differences in age, smoking index or FEV(1)/FVC between these groups. CONCLUSION: These results suggest that high antibody titres against C. pneumoniae are linked with the severity of emphysema on high resolution CT and decreased diffusing capacity to carbon monoxide. PMID- 16268910 TI - Asthma control in adults in Asia-Pacific. AB - OBJECTIVE: The Asthma Insights and Reality in Asia-Pacific (AIRIAP) survey collected detailed information on asthma severity and management in the urban centres of eight areas of the Asia-Pacific region. This study compared asthma morbidity and management practices in these areas. METHODOLOGY: Following recruitment, face-to-face interviews were completed with 2323 adults with diagnosed asthma, who had current symptoms or were using asthma medication. Comparisons between areas were made for asthma severity, asthma burden and management practices. RESULTS Asthma severity varied significantly between areas (P < 0.01), with Vietnam and mainland China reporting the most cases with severe, persistent symptoms. Severity of asthma was significantly associated with advancing age and a lower level of education in a multivariate analysis (P < 0.001). The total use of acute healthcare for asthma was significantly associated with increased asthma severity. Work absence due to asthma was highest in the Philippines (46.6%) and lowest in South Korea (7.5%). The use of inhaled corticosteroids was associated with age in a non-linear manner. There was significant variation among countries in usage of inhaled corticosteroids, from 1.3% in South Korea to 29.0% in Taiwan (P < 0.00001). A peak flow meter was owned by a total of 7.7% of respondents, and overall, 17.9% of adults had a written action plan for asthma management. CONCLUSIONS: Within the Asia-Pacific region, asthma in adults differs significantly in disease severity, management and treatment according to area of residence. International recommendations on the management of asthma are generally not being followed. PMID- 16268911 TI - Assessment of an asthma quality of life scale using item-response theory. AB - OBJECTIVE: Health-related quality of life (HRQOL) scores produced by simply summing individual item values have been criticized for lacking linearity and for not being equally discriminating across the range of scores. Differences in summed scores may depend more on their starting point on the scale rather than actual differences in the underlying dimension of HRQOL, making it difficult to judge what a particular score means. We sought to examine the usefulness of an alternative method of scoring questionnaires, item-response theory (IRT), for the clinical interpretation of a modified version of the Marks Asthma Quality of Life Questionnaire (MAQLQ-M). METHODOLOGY: Using the MAQLQ-M, we surveyed 293 adults with moderate to severe asthma, managed at two university teaching hospitals in Adelaide, South Australia. Scores obtained by usual summative Likert-type scores were compared to estimates using the partial credit method of IRT. RESULTS: We found a non-linear relationship between raw, summative scores and the IRT estimates. The departure from linearity was marked for summative scores below 3.0 and above 5.0 (range 1.0-7.0), values that included half of the study patients. Summative scoring did not produce scores at the interval level of measurement. CONCLUSION: For an equivalent difference measured in the underlying dimension of actual HRQOL using IRT, traditional summated scores showed a much smaller difference in scores at both the lower and upper end of HRQOL, than at the mid range of HRQOL. Caution should be used when interpreting HRQOL surveys scored in the usual summative manner. Advantages may be gained by using IRT as an alternative method of scoring HRQOL questionnaires. PMID- 16268912 TI - COPD: a prevalence estimation model. AB - OBJECTIVES: COPD is increasingly recognized as a leading cause of global morbidity and mortality. Prevalence estimates for COPD are generally unavailable or unreliable. Thus, a simple and valid model for estimating COPD prevalence would provide essential information for policymakers in addressing a major burden of worldwide illness. METHODOLOGY: We modelled the relationships among readily available demographic data (e.g. age, gender), smoking prevalence, and COPD prevalence based on a literature review. We also included risks of COPD from environmental pollution and associations with socioeconomic status. RESULTS: The model specifies a minimum of eight input variables to predict COPD prevalence in a given population: population by age, gender, smoking prevalence, prevalence of COPD among smokers, proportion living in rural areas, country by level of development, and exposures to environmental pollution. Actual COPD prevalence data from large population-based studies in Spain, Norway, Poland and Nepal compared favourably with the model projections (P > or = 0.10). CONCLUSION: The model is a simple tool for estimating the prevalence of COPD populations in a given region or country. Further studies are needed to prospectively validate the model and test the assumptions upon which it is based. PMID- 16268913 TI - Response to bronchodilator in infants with bronchiolitis can be predicted from wheeze characteristics. AB - OBJECTIVE: Lung sounds analysis has been used for clinical care. Our objectives were to characterize the spectral pattern of lung sounds and their relation to bronchodilator effects in acute bronchiolitis (AB). We hypothesized that patients with sinusoidal wheezes (SW) would show a more significant bronchodilator response. METHODOLOGY: We studied 22 asleep hospitalized infants (14 boys, eight girls), aged 5.2 +/- 1 months, 16 with a positive respiratory syncytial virus test, during their first 3 days after admission. Patients breathed spontaneously through a face mask connected to a pneumotachograph during normal breathing, and only target flows of 0.1 +/- 0.02 L/s were analyzed. Sounds were obtained using two contact sensors attached over both posterior lower lobes. For inspiratory and expiratory sounds, we determined the frequencies below which 25% (F25), 50% (F50), 75% (F75) and 99% (SEF99) of the spectral power between 100 and 1000 Hz was contained. We repeated the measurements 20 min after bronchodilator therapy in all patients. RESULTS: We found classic SW in 11 patients, while the other 11 had complex wheezes (CW). There were positive bronchodilator responses in 9/11 with SW and 3/11 with CW (P < 0.01). Patients who responded to salbutamol showed an increase in power at low frequencies after medication (P < 0.01), and a positive correlation between wheezing and the increase in the power spectra measured by F50 and SEF99 (P < 0.001). CONCLUSIONS: We conclude that sinusoidal and complex wheezes occur in patients with AB, that a positive response to bronchodilator is significantly more common in those with classic SW and that lung sounds analysis is a reproducible, safe and non-invasive method for assessing wheeze in infants. PMID- 16268914 TI - Inflammatory activity is still present in the advanced stages of idiopathic pulmonary fibrosis. AB - OBJECTIVE: The role of active inflammation in idiopathic pulmonary fibrosis (IPF) is controversial. A gallium-67 citrate (Ga(67) scan) is a sensitive indicator of inflammatory activity. The aim of this study was to assess the Ga(67) uptake and other markers of inflammation at different stages of IPF and to investigate its prognostic role. METHODOLOGY: Twenty-two patients (aged 66 +/- 11 years, 18 males) with IPF were monitored for a period of 6-20 months (mean 13 months). At presentation (T0), high resolution CT (HRCT) scans showed reticular opacities and traction bronchiectasis with bi-basilar and peripheral distribution in all cases. At both T0 and follow-up (T1), we measured pulmonary function (PaO(2), FVC, DLco), overall radiographic extent of fibrosis (HRCT visual score), Ga(67) uptake, serum concentrations of lactate dehydrogenase (LDH) and C-reactive protein (CRP), and erythrocyte sedimentation rate (ESR). RESULTS: All parameters showed a significant deterioration during the T0-T1 interval, though the increase in Ga(67) uptake and serum markers was not significant. Patients with Ga(67) uptake indices graded as normal or mildly increased (group I), and graded as considerably or severely increased (group II) at presentation, were compared. There was no significant difference with respect to lung function or HRCT score between the two groups at T1. Ga(67) uptake, LDH, CRP and ESR at presentation did not correlate significantly with the interval change in pulmonary function and disease extent. CONCLUSIONS: Our findings indicate that inflammatory activity in the advanced stage of IPF is still relevant, although a Ga(67) scan is not predictive of the clinical course. PMID- 16268915 TI - Protective effects of N-acetylcysteine on the peroxidative changes of rat lungs exposed to inhalation of thinners. AB - OBJECTIVE: Long-term inhalation of thinners may cause damage, both to the lungs and to other organ systems. It causes cellular damage via formation of reactive oxygen species. The lung is protected from oxidative stress by the glutathione (GSH) antioxidant system which can be augmented by the thiol drug, N acetylcysteine (NAC). This study investigated the protective effect of NAC on peroxidative changes in rat lungs exposed to inhalation of thinners for 8 weeks. METHODOLOGY: Seventy-two male Wistar albino rats were used and divided into two groups: one group inhaled only thinners (TI), while the other inhaled TI plus NAC. Rats in the TI and TI + NAC groups were divided into four subgroups (each consisting of eight rats) according to the duration of exposure to TI: 2, 4, 6 and 8 weeks. A control group (n = 7) of rats inhaled neither TI nor NAC. Malondialdehyde (MDA) and GSH levels, and superoxide dismutase (SOD) activities were determined in the lung tissues. Histopathological findings were evaluated as acute and chronic changes in the alveoli and interstitium in the TI and TI + NAC groups and compared with those in the control group. RESULTS: While tissue MDA levels in the groups inhaling TI for 4, 6 and 8 weeks were significantly higher than those in the control groups (P < 0.01, P < 0.01, P < 0.0001, respectively), GSH levels were significantly lower (P < 0.05, P < 0.01, P < 0.01, respectively). Tissue SOD activities in the groups inhaling TI for 6 and 8 weeks were significantly lower than those in the control group (P < 0.05, P < 0.01, respectively). In the TI group, MDA levels were significantly increased (P < 0.01) with increasing duration of inhalation (from the second week through to the eighth week), while GSH levels and SOD activities were significantly decreased (P < 0.01, P < 0.01). Tissue MDA levels were significantly lower in the TI + NAC groups across all inhalation periods, when compared with the TI groups (P < 0.01, P < 0.0001, P < 0.0001, P < 0.0001, respectively). Tissue GSH levels in the TI + NAC groups were significantly higher than those of the TI groups (respective values: P < 0.05, P < 0.01, P < 0.01, P < 0.0001). Tissue SOD activities in the TI + NAC groups were significantly higher than those of the TI groups (respective values: P < 0.05, P < 0.0001, P < 0.05, P < 0.0001). Pathological examinations with light microscopy did not show any beneficial effect of NAC application in terms of deferring or alleviating the negative effects of TI. CONCLUSIONS: Thinners are agents that cause imbalance between oxidants and antioxidants produced by aerobic cellular systems. This imbalance between oxidant and antioxidant systems is decreased by the effect of NAC. However, ultrastructural studies may be needed to substantiate this evidence morphologically, as light microscopy was inconclusive. PMID- 16268916 TI - Pulmonary artery pressure and blood flow as predictors of outcome from lung cancer resection. AB - OBJECTIVE: Pulmonary hypertension is a putative risk factor for lung resection but catheter measurements are invasive. The aim of the present study was to assess prediction of mortality and complications from lung resection by Doppler echocardiographic estimates of pulmonary artery pressure (PAp) and soluble gas uptake estimates of effective pulmonary blood flow (Q(RB)). METHODOLOGY: In 33 lung cancer patients, resting PAp (sys) and Q(RB) were measured preoperatively. In 13 patients, supine exercise estimates of PAp (sys) were also made and in five patients catheter PAp estimates were made. RESULTS: Baseline PAp (sys) was 35 +/- 5 mmHg in four patients who died and 35 +/- 5 mmHg in 27 survivors. Post-exercise PAp (sys) was 58 +/- 11 mmHg in non-survivors and 60 +/- 11 mmHg in survivors (both not significant). Resting Q(RB) was 3.9 +/- 0.35 L/min in two non-survivors compared with 4.7 +/- 0.90 L/min in 22 survivors (P = 0.12) and was significantly lower in those experiencing complications. Regression analysis showed no significant relationship between resting or post-exercise PAp and mortality, although low Q(RB) tended towards predicting mortality (P = 0.07). There were also trends for higher PAp (sys) to predict respiratory and cardiac complications (P = 0.08) and for lower Q(RB) to predict unspecified or surgical complications. CONCLUSION: PAp and Q(RB) did not predict outcome from lung resection better than baseline respiratory function indices. PMID- 16268918 TI - Frozen section of pleural biopsies at medical thoracoscopy assists in correctly identifying benign disease. AB - OBJECTIVES: Medical thoracoscopy and thoracoscopic talc poudrage (TTP) are accepted procedures in the management of pleural effusions. The relative merits of TTP compared with pleurodesis via intercostal catheter (ICC) continue to be debated. However, of the two procedures, only medical thoracoscopy allows both tissue diagnosis and pleurodesis to be achieved reliably in one procedure. The aim of this study was to assess the feasibility and accuracy of using frozen section analysis of samples taken during medical thoracoscopy to assist the thoracoscopist's decision to complete the procedure with a TTP. METHODOLOGY: Twenty patients with undiagnosed pleural effusions after at least one diagnostic pleurocentesis underwent medical thoracoscopy and biopsy. RESULTS: Frozen sections were easily performed within the timeframe of medical thoracoscopy. The final diagnosis based on paraffin sections was malignant in 10 cases and benign in 10 cases. Frozen section at the time of thoracoscopy (before TTP) correctly identified nine of 10 cases as being benign and six of 10 cases as malignant. In the malignant group, reasons for incorrect identification as benign were sampling from superficial benign adipose tissue overlying the malignant deposits, difficult access to the most involved parts of the pleura and intense cellular infiltrate initially thought to be benign. CONCLUSION: Frozen sections taken during medical thoracoscopy have the potential to facilitate decision-making prior to pleurodesis, particularly for accurate identification of benign histology on thoracoscopic pleural biopsies, in order that pleurodesis is not performed unnecessarily. PMID- 16268917 TI - Gemcitabine and carboplatin in the treatment of locally advanced and metastatic non-small cell lung cancer. AB - OBJECTIVE: The aim of the study was to evaluate the response, survival advantage and toxicity profile of gemcitabine-carboplatin combination cytotoxic chemotherapy in patients with locally advanced and metastatic non-small cell lung cancer (NSCLC). METHODOLOGY: Patients who received gemcitabine-carboplatin combination chemotherapy over a 2.5-years period were analyzed. Carboplatin at a dose of 5 mg/mL/min (area under the concentration-time curve) was given on day 1 and gemcitabine (1000 mg/m(2)) on days 1 and 8, every 3 weeks. RESULTS: Of 49 chemotherapy-naive patients (median age, 62 years) who received this treatment, 57% were males, 12% had stage IIIa, 39% stage IIIb and 49% metastatic disease. The Eastern Cooperative Oncology Group (ECOG) performance status of 70% of the patients was 1 at the time of commencement of chemotherapy and 2 for the remaining 30% of patients. The overall response rate, based on 33 evaluable patients, was 27.3%. The response rate was not affected by age, stage of disease or performance status. The median survival was 9 months. Median survival among patients with an ECOG performance status of 1 was 11 months, as compared with 4 months for patients with an ECOG performance status of 2 (P < 0.001). Toxicity was generally well tolerated and there were no treatment-related deaths. CONCLUSIONS: Gemcitabine-carboplatin combination chemotherapy is an effective and well-tolerated cytotoxic regimen among Malaysian patients with advanced NSCLC. A performance status of 1 or less was associated with a better survival. PMID- 16268919 TI - The prevalence of acute response to bronchodilator in pulmonary lymphangioleiomyomatosis. AB - OBJECTIVE: The objective of this study was to evaluate the prevalence of acute bronchodilator responses in patients who were evaluated for pulmonary lymphangioleiomyomatosis. METHODOLOGY: This study was a retrospective review of the medical records of 50 patients, who were assessed for lymphangioleiomyomatosis from 1978 to 2002 at Mayo Clinic in Rochester, Minnesota, and Jacksonville, Florida, USA. Each medical record was reviewed for pertinent clinical and pulmonary function test data, including results for lung volume and flow, diffusing capacity and response to bronchodilator. RESULTS: At initial testing, the mean FEV(1)/FVC ratio for the whole group was 60%; 35 patients (70%) had airflow obstruction; five patients (10%) had only restriction; and 10 (20%) had neither restriction nor obstruction. Three patients (6%) showed a positive response to bronchodilator at initial testing and only one patient showed a positive response at 1 year. Pulmonary function testing at 1-year follow up was available in a subgroup of this cohort (23 patients; 46%), and only one patient (2%) had a positive response to bronchodilator. Although there was a statistically significant decrease in the FEV(1)/FVC ratio (from 64% to 62%; P = 0.02), no other differences were found at 1-year follow up. CONCLUSIONS: In its advanced state, lymphangioleiomyomatosis produces irreversible airflow obstruction. A positive response to bronchodilator was unusual. Other measures of pulmonary function showed little or no change over a 12-month period. PMID- 16268920 TI - Prolonged survival after talc poudrage for malignant pleural mesothelioma: case series. AB - OBJECTIVE: Malignant pleural mesothelioma is a fatal disease with a mean life expectancy of 6-12 months. Since 1982, we have performed thoracoscopic talc poudrage (TTP) as a primary therapy in mesothelioma patients presenting with pleural effusion. As the survival data for our patients surpassed that of many published series, the patient data was analyzed to determine whether talc poudrage can be considered as a contemporary palliative option. METHODOLOGY: We reviewed all 26 patients with a final diagnosis of malignant pleural mesothelioma from our prospective database of 228 consecutive patients who received thoracoscopy from the same physician for recurrent symptomatic pleural effusion. Patients were followed up until their death. RESULTS: Mean survival after TTP was 23.8 +/- 16.3 months (median 19.4, range 2.9-68). Pleurodesis palliated dyspnoea in all patients. No perioperative deaths and one postoperative complication (pneumonia) occurred. Mean hospital stay was 3.9 +/- 2.7 days. CONCLUSION: TTP remains a safe, low-morbidity, inexpensive primary palliative treatment option for malignant pleural mesothelioma and a valid control arm option for therapeutic trials. TTP is ideal for patients who wish to avoid thoracotomy, long hospital stays and morbidity from multimodality therapy. Prospective randomized studies are needed to compare quality of life and survival after talc poudrage and other therapies. PMID- 16268921 TI - Leptospirosis and its pulmonary complications. AB - OBJECTIVE: The aim of this study was to assess the incidence and factors associated with pulmonary complications of leptospirosis. METHODOLOGY: In a retrospective study, patients with a definite diagnosis of leptospirosis following a 6-week period of severe flooding in Hadyai city, Thailand, were reviewed. Pulmonary complications of leptospirosis were defined as the occurrence of respiratory symptoms and an abnormal CXR. The clinical and laboratory test results for patients with and without pulmonary complications were compared. RESULTS: Among the 157 patients with leptospirosis, eight patients had pulmonary complications. Three patients had acute renal failure (ARF) and pulmonary oedema. One patient had ARF and adult respiratory distress syndrome (ARDS). Two patients had ARF, congestive heart failure and pulmonary oedema. One patient had congestive heart failure and pulmonary oedema. One patient had only ARF. Factors associated with pulmonary complications were delayed antibiotic treatment and thrombocytopenia (platelet count < 100 x 10(9)/L). Three patients developed adult respiratory distress syndrome and one died from respiratory failure. CONCLUSIONS: Pulmonary complications and death occur in a low percentage of patients with leptospirosis. Delayed antibiotic treatment and thrombocytopenia are risk factors for the development of pulmonary involvement in leptospirosis. PMID- 16268922 TI - Clinicopathological analysis of lung cancer resembling malignant pleural mesothelioma. AB - OBJECTIVE: The aim of this study was to evaluate the clinicopathological features of lung cancer resembling malignant pleural mesothelioma. METHODOLOGY: The seven patients studied had tumours showing an extensive pleural growth pattern, and were chosen from 1516 lung cancer patients diagnosed at two affiliated hospitals over a 17-year period. RESULTS: Histologically, five of these lung cancers were adenocarcinomas, one was a small cell carcinoma, and one a large cell carcinoma. Tumour markers such as carcinoembryonic antigen (CEA) were elevated in six patients at admission. However, a clinical diagnosis was difficult and these cancers could only be distinguished from malignant pleural mesothelioma by thoracentesis in two cases, pleural biopsy in two, thoracotomy in two, and in case 7 at autopsy. Treatment consisted of chemotherapy or radiation therapy in four patients, but with little clinical effect. The mean survival time was 6.7 months, which is much less than for true malignant pleural mesothelioma. CONCLUSIONS: Lung cancer resembling malignant pleural mesothelioma is most frequently observed in patients with adenocarcinoma, which is suspected to occur peripherally. To distinguish pseudomesotheliomatous carcinoma from malignant pleural mesothelioma in patients with diffuse pleural thickening and effusion, requires adequate tissue sampling by thoracotomy or video-assisted thoracoscopic surgery and a panel of immunohistochemical stains. PMID- 16268923 TI - Cranial optic nerve involvements in patients with severe COPD. AB - OBJECTIVE: The relationship between neuropathy and increased morbidity in patients with COPD is clear, but few studies have assessed cranial neuropathies, especially optic nerve involvement, in COPD patients. We evaluated peripheral involvement of the optic nerve and determined factors influencing this condition in patients with severe COPD. METHODOLOGY: Twenty-eight patients, mean age 59.4 +/- 9.4 years, diagnosed with severe stable COPD according to the GOLD criteria, and 20 age- and gender-matched healthy individuals, mean age 55.6 +/- 8.5 years, were included in the study. All subjects underwent visual evoked potential (VEP) assessment together with detailed clinical and laboratory examination to exclude concurrent risk factors for neuropathy. RESULTS: VEP assessment showed significant abnormalities in COPD patients (82.1%) (commonly amplitude abnormalities) when compared with healthy controls. CONCLUSIONS: The optic nerve is often involved in patients with severe COPD, possibly as part of a polyneuropathy, and this is related to acidosis, hypercarbia and airway obstruction, independent of disease duration, smoking and age. These results should be taken into consideration when determining management strategies for these patients. PMID- 16268924 TI - The feasibility of nitric oxide delivery with high frequency jet ventilation. AB - A 27-year-old female with acute monocytic leukaemia (M5) developed acute respiratory distress syndrome (ARDS). Because of refractory hypoxaemia and severe barotrauma during conventional mechanical ventilation, the patient was switched to high frequency jet ventilation (HFJV) as a salvage therapy. Her refractory hypoxaemia improved temporarily but worsened again. Approval of the Institutional Review Board and the Food and Drug Administration was obtained to use nitric oxide (NO) with HFJV on a compassionate basis considering the grave situation. The NO delivery system was connected to the secondary flow circuit of the HFJV immediately after the humidifier. We measured the concentration of NO at least every hour by inserting a 7-Fr catheter connected to a McNeill analyzer into the endotracheal tube. Despite improvement of oxygenation, the patient's respiratory status deteriorated further and she died. In this case we were able to achieve reliable and constant levels of NO. The purpose of this report is to discuss the technical aspects and pitfalls of this method. PMID- 16268925 TI - Microscopic polyangiitis with unusual lung involvement. AB - Microscopic polyangiitis (MPA) is a non-granulomatous, systemic and small vessel vasculitis accompanied by segmental necrotizing glomerulonephritis with no evidence of other small vessel disease. We report a patient with weakness, fever, and arthralgia whose CXR and thoracic CT showed widespread nodular infiltration. His proteinase-3 anti-neutrophilic cytoplasmic antibody (c-ANCA) was positive. The serum creatinine was increased and haematuria subsequently developed. Renal biopsy revealed a focal segmental necrotizing glomerulonephritis which was compatible with MPA. He was treated with high-dose corticosteroid and cyclophosphamide. Because of a worsening CXR and hypoxaemia, mechanical ventilation was applied. Despite this he died of respiratory failure following 20 days of treatment. Nodular infiltration is an unusual radiological pattern in patients with MPA and is the reason for this report. PMID- 16268926 TI - Chronic pulmonary scedosporiosis simulating aspergillosis. AB - Scedosporium apiospermum, a ubiquitously distributed saprophyte, is emerging as an important pathogen in immunocompromised patients. We describe a 72-year-old patient with chronic S. apiospermum infection of the lung simulating aspergilloma. His medical history was unremarkable except that he had undergone partial lung resection as a treatment for pulmonary tuberculosis several decades previously. He had no underlying immunosuppressive conditions. This patient illustrates that pulmonary scedosporiosis is not confined to immunocompromised patients and that the clinical presentation may be indistinguishable from that of aspergilloma. PMID- 16268927 TI - Intralobar sequestration with tuberculous infection confined to the sequestrated lung. AB - A 22-year-old female was referred to the hospital suffering from a persistent, non-productive cough and repeated exacerbations of a right lower zone infiltrate suggestive of pulmonary sequestration. Angiography revealed an aberrant artery that originated from the left side of the descending aorta, crossed the aorta anteriorly, flowed through the right lower lobe and drained into the right inferior pulmonary vein. Right lower lobectomy was conducted by open thoracotomy. Gross examination revealed mucoid impaction of bronchi in the sequestration. Microscopy demonstrated a clear boundary between sequestrated and normal lung, as well as caseating epitheloid granulomas confined to the sequestration. As the aberrant artery fed both sequestrated and adjacent normal lung and the boundary did not involve the pleura, the lesion was classified as a Pryce's type II intralobar sequestration. No acid-fast bacilli were observed, but homogenates of the sequestration were positive for Mycobacterium tuberculosis by polymerase chain reaction. The patient was treated with isoniazid and rifampicin daily for 9 months, and 6 years later her clinical status remained excellent. Tuberculosis confined to a sequestration is extremely rare and characteristics of the aberrant artery suggested this intralobar sequestration was likely to be congenital in origin. PMID- 16268928 TI - Thymic carcinoma originating from the mid-posterior mediastinum. AB - An unusual thymic carcinoma in a 74-year-old woman is described. Initial chest CT revealed a mass at the mid-posterior mediastinum. Transbronchial fine needle biopsy of the mass failed to provide a definite diagnosis. The mass was treated as a malignant mediastinal tumour, and chemoradiotherapy was performed as initial treatment. The patient died 5 years after receiving primary treatment. The results of postmortem microscopic examination, including immunohistochemical study with CD5 antibody, were consistent with thymic carcinoma. This case is interesting in that the mid-posterior mediastinum is the site where thymic carcinoma is least likely to originate. PMID- 16268929 TI - What is a life worth? Robustness of VSL values from contingent valuation surveys. AB - The value of a statistical life (VSL) is a key input for estimating the benefits of policies that save lives. Several recent studies have obtained estimates of the VSL from contingent valuation surveys, i.e., by asking people to say how much they would pay to reduce their risk of dying. This article examines statistical factors that may influence the estimates of the VSL obtained from such surveys. We examine the importance of distributional assumptions, the choice of the welfare statistics of interest, the procedure for computing them, outliers, undesirable response effects, and internal validity of the willingness-to-pay (WTP) responses. We illustrate the importance of these factors using dichotomous choice and open-ended WTP data from four recent contingent valuation surveys. PMID- 16268930 TI - Estimating the health impacts of tobacco harm reduction policies: a simulation modeling approach. AB - With adult smoking prevalence rates declining too slowly to reach national objectives, opinion leaders are considering policies to improve tobacco-related outcomes by regulating the composition of cigarettes to be (1) less harmful and/or (2) less addictive. Because harm reduction efforts may actually encourage higher cigarette consumption by promoting a safer image, and addictiveness reduction may increase the harmfulness of cigarettes by encouraging compensatory smoking behaviors, policymakers must consider the tradeoffs between these two approaches when proposing legislation to control cigarette content. To estimate health impacts, we developed a dynamic computer model simulating changes in the age- and gender-specific smoking behaviors of the U.S. population over time. Secondary data for model parameters were obtained from publicly available sources. Population health impacts were measured as change in smoking prevalence and the change in cumulative quality-adjusted life-years (QALYs) in the U.S. population over 75 years. According to the risk-use threshold matrix generated by the simulation, modifying cigarettes to reduce their harmfulness and/or addictiveness could result in important gains to the nation's health. Addictiveness reduction efforts producing a 60% improvement in smoking behavior change probabilities would produce a net gain in population health at every plausible level of increase of smoking-related harm that was modeled. A 40% reduction in smoking-related harm would produce a net QALY gain at every level of behavior change considered. This research should prove useful to policymakers as they contemplate giving the FDA the authority to regulate the composition of cigarettes. PMID- 16268931 TI - Risk perception of the "mad cow disease" in France: determinants and consequences. AB - Since 1996, when bovine spongiform encephalopathy (BSE) was assessed as a possible human transmissible disease, a variant of Creutzfeldt-Jakob disease (vCJD), French people have entered into a long period of fear and avoidance of beef and bovine byproducts, which produced an unprecedented collapse in the beef market. This article deals with the perceived risk of the "mad cow disease" (MCD) in the French general population. Two surveys were conducted on a representative sample of the adult population, the first one in 2000 during the peak of the crisis and the second one 13 months later in a quieter period. The main assumption we made was that changes in beef consumption are strongly related to the perceived risk of MCD, which we defined as people's cognitive and affective responses to hazard. Our objective was to identify the determinants and consequences of this perceived risk and to compare them in different sociopolitical contexts. The results issued from a bivariate and multivariate analysis show that: (i) the distribution of most of the variables significantly related to the perceived risk identified in the first survey had changed in the second survey, in relation with the reduction of worry and the resumption of national beef consumption; (ii) the propensity for self-protection through avoiding or ceasing beef eating was more related to feelings of worry than to subjective vCJD risk assessments; and (iii) the main determinant of less avoidance to beef products was the preference for beef, a feeling identified prior to emergence of the risk of MCD, remaining unchanged in various contexts. PMID- 16268932 TI - Optimal tracking and testing of U.S. and Canadian herds for BSE: a value-of information (VOI) approach. AB - The U.S. Department of Agriculture (USDA) tests a subset of cattle slaughtered in the United States for bovine spongiform encephalitis (BSE). Knowing the origin of cattle (U.S. vs. Canadian) at testing could enable new testing or surveillance policies based on the origin of cattle testing positive. For example, if a Canadian cow tests positive for BSE, while no U.S. origin cattle do, the United States could subject Canadian cattle to more stringent testing. This article illustrates the application of a value-of-information (VOI) framework to quantify and compare potential economic costs to the United States of implementing tracking cattle origins to the costs of not doing so. The potential economic value of information from a tracking program is estimated to exceed its costs by more than five-fold if such information can reduce future losses in export and domestic markets and reduce future testing costs required to reassure or win back customers. Sensitivity analyses indicate that this conclusion is somewhat robust to many technical, scientific, and market uncertainties, including the current prevalence of BSE in the United States and/or Canada and the likely reactions of consumers to possible future discoveries of BSE in the United States and/or Canada. Indeed, the potential value of tracking information is great enough to justify locating and tracking Canadian cattle already in the United States when this can be done for a reasonable cost. If aggressive tracking and testing can win back lost exports, then the VOI of a tracking program may increase to over half a billion dollars per year. PMID- 16268933 TI - An interagency comparison of screening-level risk assessment approaches. AB - Approaches to risk assessment have been shown to vary among regulatory agencies and across jurisdictional boundaries according to the different assumptions and justifications used. Approaches to screening-level risk assessment from six international agencies were applied to an urban case study focusing on benzo[a]pyrene (B[a]P) exposure and compared in order to provide insight into the differences between agency methods, assumptions, and justifications. Exposure estimates ranged four-fold, with most of the dose stemming from exposure to animal products (8-73%) and plant products (24-88%). Total cancer risk across agencies varied by two orders of magnitude, with exposure to air and plant and animal products contributing most to total cancer risk, while the air contribution showed the greatest variability (1-99%). Variability in cancer risk of 100-fold was attributed to choices of toxicological reference values (TRVs), either based on a combination of epidemiological and animal data, or on animal data. The contribution and importance of the urban exposure pathway for cancer risk varied according to the TRV and, ultimately, according to differences in risk assessment assumptions and guidance. While all agency risk assessment methods are predicated on science, the study results suggest that the largest impact on the differential assessment of risk by international agencies comes from policy and judgment, rather than science. PMID- 16268934 TI - Cellular telephones and driving performance: the effects of attentional demands on motor vehicle crash risk. AB - This study examines the effects of conversation mode and split-attention communication training on driving performance. The study is based on an experiment where drivers with and without communication training (pilots vs. nonpilots) completed a simulated driving course while involved in one of three conversation modes: no conversation, conversation with passenger, or conversation on a hands-free cellular telephone. Results indicate that cellular telephone conversations consume more attention and interfere more with driving than passenger conversations. Cell phone conversations lack the nonverbal cues available during close-contact conversations and conversation participants expend significant cognitive resources to compensate for the lack of such cues. The results also demonstrate that communication training may reduce the hazardous effects of cell phone conversations on driving performance. PMID- 16268935 TI - Why has safety improved at rail-highway grade crossings? AB - The number of collisions and fatalities at rail-highway intersections in the United States has declined significantly over the past 30 years, despite considerable increases in the volume of rail and highway traffic. This article disaggregates the improvement into its constituent causes. Negative binomial regressions are conducted on a pooled data set for 49 states from 1975 to 2001. The analysis concludes that about two-fifths of the decrease is due to factors such as reduced drunk driving and improved emergency medical response that have improved safety on all parts of the highway network. The installation of gates and/or flashing lights accounts for about a fifth of the reduction. The development in the 1970s and early 1980s of the Operation Lifesaver public education campaign, and the installation of additional lights on locomotives in the mid 1990s, each led to about a seventh of the reduction. Finally, about a tenth is due to closure of crossings resulting from line abandonments or consolidation of little-used crossings. PMID- 16268936 TI - A benefit-cost analysis of retrofitting diesel vehicles with particulate filters in the Mexico City metropolitan area. AB - In the Mexico City metropolitan area, poor air quality is a public health concern. Diesel vehicles contribute significantly to the emissions that are most harmful to health. Harmful diesel emissions can be reduced by retrofitting vehicles with one of several technologies, including diesel particulate filters. We quantified the social costs and benefits, including health benefits, of retrofitting diesel vehicles in Mexico City with catalyzed diesel particulate filters, actively regenerating diesel particulate filters, or diesel oxidation catalysts, either immediately or in 2010, when capital costs are expected to be lower. Retrofit with either type of diesel particulate filter or an oxidation catalyst is expected to provide net benefits to society beginning immediately and in 2010. At current prices, retrofit with an oxidation catalyst provides greatest net benefits. However, as capital costs decrease, retrofit with diesel particulate filters is expected to provide greater net benefits. In both scenarios, retrofit of older, dirtier vehicles that circulate only within the city provides greatest benefits, and retrofit with oxidation catalysts provides greater health benefits per dollar spent than retrofit with particulate filters. Uncertainty about the magnitude of net benefits of a retrofit program is significant. Results are most sensitive to values used to calculate benefits, such as the concentration-response coefficient, intake fraction (a measure of exposure), and the monetary value of health benefits. PMID- 16268937 TI - Risk management and the precautionary principle: a fuzzy logic model. AB - The aim of this article is to illustrate a procedure for applying the precautionary principle within a strategy for reducing the possibility of underestimating the effective risk caused by a phenomenon, product, or process, and of adopting insufficient risk reduction measures or overlooking their need. We start by simply defining risk as the product between the numerical expression of the adverse consequences of an event and the likelihood of its occurrence or the likelihood that such consequences will occur. Uncertainty in likelihood estimates and several key concepts inherent to the precautionary principle, such as sufficient certainty, prevention, and desired level of protection, are represented as fuzzy sets. The strategy described may be viewed as a simplified example of a precautionary decision process that has been chiefly conceived as a theoretical contribution to the debate concerning the precautionary principle, the quantification of its application, and the formal approach to such problems. PMID- 16268938 TI - Benchmark analysis: shopping with proper confidence. AB - We discuss the issue of using benchmark doses for quantifying (excess) risk associated with exposure to environmental hazards. The paradigm of low-dose risk estimation in dose-response modeling is used as the primary application scenario. Emphasis is placed on making simultaneous inferences on benchmark doses when data are in the form of proportions, although the concepts translate easily to other forms of outcome data. PMID- 16268939 TI - A numerical solution to the nonhomogeneous two-stage MVK model of cancer. AB - In this article, we describe a straightforward method for solving the probability of at least one malignant cell by time t, and the associated hazard function, in the general (i.e., nonhomogeneous) two-stage Moolgavkar-Venzon-Knudson (MVK) model of cancer. The method consists of solving four coupled ordinary differential equations derived from the Kolmogorov backward equations for this process. The relationship of this method to previously proposed solutions is discussed. PMID- 16268940 TI - Developing policy in the face of scientific uncertainty: interpreting 0.3 microT or 0.4 microT cutpoints from EMF epidemiologic studies. AB - There has been considerable scientific effort to understand the potential link between exposures to power-frequency electric and magnetic fields (EMF) and the occurrence of cancer and other diseases. The combination of widespread exposures, established biological effects from acute, high-level exposures, and the possibility of leukemia in children from low-level, chronic exposures has made it both necessary and difficult to develop consistent public health policies. In this article we review the basis of both numeric standards and precautionary based approaches. While we believe that policies regarding EMF should indeed be precautionary, this does not require or imply adoption of numeric exposure standards. We argue that cutpoints from epidemiologic studies, which are arbitrarily chosen, should not be used as the basis for making exposure limits due to a number of uncertainties. Establishment of arbitrary numeric exposure limits undermines the value of both the science-based numeric EMF exposure standards for acute exposures and precautionary approaches. The World Health Organization's draft Precautionary Framework provides guidance for establishing appropriate public health policies for power-frequency EMF. PMID- 16268941 TI - Mesothelioma: risk apportionment among asbestos exposure sources. AB - The mesothelioma epidemic in the United States, which peaked during the 2000-2004 period, can be traced to high-level asbestos exposures experienced by males in occupational settings prior to the full recognition of the disease-causing potential of asbestos and the establishment of enforceable asbestos exposure limits by the Occupational Safety and Health Administration (OSHA) in 1971. Many individuals diagnosed with mesothelioma where asbestos has been identified as a contributing cause of the disease have filed claims seeking compensation from asbestos settlement trusts or through the court system. An individual with mesothelioma typically has been exposed to asbestos in more than one setting and from more than one asbestos product. Apportioning risk for mesothelioma among contributing factors is an ongoing problem faced by occupational disease compensation boards, juries, parties responsible for paying damages, and currently by the U.S. Senate in its efforts to formulate a bill establishing an asbestos settlement trust. In this article we address the following question: If an individual with mesothelioma where asbestos has been identified as a contributing cause were to be compensated for his or her disease, how should that compensation be apportioned among those responsible for the asbestos exposures? For the purposes of apportionment, we assume that asbestos is the only cause of mesothelioma and that every asbestos exposure contributes, albeit differentially, to the risk. We use an extension of the mesothelioma risk model initially proposed in the early 1980s to quantify the contribution to risk of each exposure as a percentage of the total risk. The percentage for each specific discrete asbestos exposure depends on the start and end dates, the intensity, and the asbestos fiber type for the exposure. We provide justification for the use of the mesothelioma risk model for apportioning risk and discuss how to assess uncertainty associated with its application. PMID- 16268942 TI - Exposure reconstruction for the TCDD-exposed NIOSH cohort using a concentration- and age-dependent model of elimination. AB - Recent studies demonstrating a concentration dependence of elimination of 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) suggest that previous estimates of exposure for occupationally exposed cohorts may have underestimated actual exposure, resulting in a potential overestimate of the carcinogenic potency of TCDD in humans based on the mortality data for these cohorts. Using a database on U.S. chemical manufacturing workers potentially exposed to TCDD compiled by the National Institute for Occupational Safety and Health (NIOSH), we evaluated the impact of using a concentration- and age-dependent elimination model (CADM) (Aylward et al., 2005) on estimates of serum lipid area under the curve (AUC) for the NIOSH cohort. These data were used previously by Steenland et al. (2001) in combination with a first-order elimination model with an 8.7-year half-life to estimate cumulative serum lipid concentration (equivalent to AUC) for these workers for use in cancer dose-response assessment. Serum lipid TCDD measurements taken in 1988 for a subset of the cohort were combined with the NIOSH job exposure matrix and work histories to estimate dose rates per unit of exposure score. We evaluated the effect of choices in regression model (regression on untransformed vs. ln-transformed data and inclusion of a nonzero regression intercept) as well as the impact of choices of elimination models and parameters on estimated AUCs for the cohort. Central estimates for dose rate parameters derived from the serum-sampled subcohort were applied with the elimination models to time-specific exposure scores for the entire cohort to generate AUC estimates for all cohort members. Use of the CADM resulted in improved model fits to the serum sampling data compared to the first-order models. Dose rates varied by a factor of 50 among different combinations of elimination model, parameter sets, and regression models. Use of a CADM results in increases of up to five-fold in AUC estimates for the more highly exposed members of the cohort compared to estimates obtained using the first-order model with 8.7-year half-life. This degree of variation in the AUC estimates for this cohort would affect substantially the cancer potency estimates derived from the mortality data from this cohort. Such variability and uncertainty in the reconstructed serum lipid AUC estimates for this cohort, depending on elimination model, parameter set, and regression model, have not been described previously and are critical components in evaluating the dose-response data from the occupationally exposed populations. PMID- 16268943 TI - Benchmark dose of lead inducing anemia at the workplace. AB - To estimate the critical dose of lead inducing anemia in humans, the effects of lead on hemoglobin (Hb) and hematocrit (Hct) levels and red blood cell (RBC) count were examined in 388 male lead-exposed workers with blood lead (BPb) levels of 0.05-5.5 (mean 1.3) micromol/L by using the benchmark dose (BMD) approach. The BPb level was significantly related to Hb (regression coefficient beta=-0.276), RBC (beta=-11.35), and Hct (beta=-0.563) among the workers (p < 0.001) when controlling for age and working status. The average BPb levels were significantly higher in the workers with anemia (1.85 micromol/L), based on the WHO criteria, than in those without anemia (1.26 micromol/L). The benchmark dose levels of BPb (i.e., lower 95% confidence limits of BMD), calculated from the K-power model set at an abnormal probability of 5% in unexposed workers and an excess risk of 5% in exposed workers were estimated to be 0.94 micromol/L (19.5 microg/dl) for Hb, 0.94 micromol/L (19.4 microg/dl) for RBC, and 1.43 micromol/L (29.6 microg/dl) for Hct. These findings suggest that reduction in hematopoietic indicators may be initiated at BPbs below the level currently considered without effect. PMID- 16268944 TI - Using Bayesian networks to model expected and unexpected operational losses. AB - This report describes the use of Bayesian networks (BNs) to model statistical loss distributions in financial operational risk scenarios. Its focus is on modeling "long" tail, or unexpected, loss events using mixtures of appropriate loss frequency and severity distributions where these mixtures are conditioned on causal variables that model the capability or effectiveness of the underlying controls process. The use of causal modeling is discussed from the perspective of exploiting local expertise about process reliability and formally connecting this knowledge to actual or hypothetical statistical phenomena resulting from the process. This brings the benefit of supplementing sparse data with expert judgment and transforming qualitative knowledge about the process into quantitative predictions. We conclude that BNs can help combine qualitative data from experts and quantitative data from historical loss databases in a principled way and as such they go some way in meeting the requirements of the draft Basel II Accord (Basel, 2004) for an advanced measurement approach (AMA). PMID- 16268945 TI - Competing failure risk analysis using evidence theory. AB - Safety systems are important components of high-consequence systems that are intended to prevent the unintended operation of the system and thus the potentially significant negative consequences that could result from such an operation. This presentation investigates and illustrates formal procedures for assessing the uncertainty in the probability that a safety system will fail to operate as intended in an accident environment. Probability theory and evidence theory are introduced as possible mathematical structures for the representation of the epistemic uncertainty associated with the performance of safety systems, and a representation of this type is illustrated with a hypothetical safety system involving one weak link and one strong link that is exposed to a high temperature fire environment. Topics considered include (1) the nature of diffuse uncertainty information involving a system and its environment, (2) the conversion of diffuse uncertainty information into the mathematical structures associated with probability theory and evidence theory, and (3) the propagation of these uncertainty structures through a model for a safety system to obtain representations in the context of probability theory and evidence theory of the uncertainty in the probability that the safety system will fail to operate as intended. The results suggest that evidence theory provides a potentially valuable representational tool for the display of the implications of significant epistemic uncertainty in inputs to complex analyses. PMID- 16268946 TI - Improbability of igneous intrusion promoting a critical event in spent nuclear fuel disposed in unsaturated tuff. AB - In their regulations, the U.S. Environmental Protection Agency and the U.S. Nuclear Regulatory Commission permit the omission of features, events, or processes with probabilities of <10(-4) in 10(4) yr (e.g., a constant frequency of <10(-8) per yr) in assessments of the performance of radioactive waste disposal systems. Igneous intrusion (or "volcanism") of a geologic repository at Yucca Mountain for radioactive waste is one disruptive event that has a probability with a range of uncertainty that straddles this regulatory criterion and is considered directly in performance assessment calculations. A self sustained nuclear chain reaction (or "criticality") is another potentially disruptive event to consider, although it was never found to be important when evaluating the efficacy of radioactive waste disposal since the early 1970s. The thesis of this article is that the consideration of the joint event--volcanism and criticality--occurring in any 10,000-year period following closure can be eliminated from performance calculations at Yucca Mountain. The probability of the joint event must be less than the fairly well-accepted but low probability of volcanism. Furthermore, volcanism does not "remove" or "fail" existing hydrologic or geochemical constraints at Yucca Mountain that tend to prevent concentration of fissile material. Prior to general corrosion failure of waste packages, the mean release of fissile mass caused by a low-probability, igneous intrusive event is so small that the probability of a critical event is remote, even for highly enriched spent nuclear fuel owned by the U.S. Department of Energy. After widespread failure of packages occurs, the probability of the joint event is less than the probability of criticality because of the very small influence of volcanism on the mean fissile mass release. Hence, volcanism plays an insignificant role in inducing criticality over any 10(4)-yr period. We also argue that the Oklo reactors serve as a natural analogue and provide a rough bound on probability of criticality given favorable hydrologic or geochemical conditions on the scale of the repository that is less than 0.10. Because the product of this bound with the probability of volcanism represents the probability of the joint event and the product is less than 10(-4) in 10(4) yr, consideration of the joint event can be eliminated from performance calculations. PMID- 16268947 TI - Understanding organizational safety using value-focused thinking. AB - Decision analysis is recognized as the right way to make risk management decisions, using probabilistic techniques to assess the accident risk. It is also accepted that the decisions that individuals in the organization make affect the likelihood of an accident and thus managerial and organizational factors should be included in the risk modeling process. However, decision analytic techniques have not been used to understand the decisions that are made by these individuals. The initial domain for this research is marine transportation. We use the framework of value-focused thinking in order to understand safety decisions made within our research partner organization, a major domestic oil tanker operator. We describe the results of interviews held with managers and employees from this organization. Through these interviews, we sought to understand the values these experts apply in their roles within the organization and the objectives they seek to achieve to contribute to its overall safety performance. The end result is a framework that not only portrays the fundamental objectives of safe operations for various roles in the organization, but also interconnects these different decision contexts. We believe that this approach is fundamentally different from those used in previous work and that this is an interesting application of value-focused thinking. PMID- 16268948 TI - Genetic diversity of invasive species in the Great Lakes versus their Eurasian source populations: insights for risk analysis. AB - Combining DNA variation data and risk assessment procedures offers important diagnostic and monitoring tools for evaluating the relative success of exotic species invasions. Risk assessment may allow us to understand how the numbers of founding individuals, genetic variants, population sources, and introduction events affect successful establishment and spread. This is particularly important in habitats that are "hotbeds" for invasive species--such as the North American Great Lakes. This study compares genetic variability and its application to risk assessment within and among three Eurasian groups and five species that successfully invaded the Great Lakes during the mid 1980s through early 1990s; including zebra and quagga mussels, round and tubenose gobies, and the ruffe. DNA sequences are compared from exotic and native populations in order to evaluate the role of genetic diversity in invasions. Close relatives are also examined, since they often invade in concert and several are saline tolerant and are likely to spread to North American estuaries. Results show that very high genetic diversity characterizes the invasions of all five species, indicating that they were founded by very large numbers of propagules and underwent no founder effects. Genetic evidence points to multiple invasion sources for both dreissenid and goby species, which appears related to especially rapid spread and widespread colonization success in a variety of habitats. In contrast, results show that the ruffe population in the Great Lakes originated from a single founding population source from the Elbe River drainage. Both the Great Lakes and the Elbe River populations of ruffe have similar genetic diversity levels--showing no founder effect, as in the other invasive species. In conclusion, high genetic variability, large numbers of founders, and multiple founding sources likely significantly contribute to the risk of an exotic species introduction's success and persistence. PMID- 16268949 TI - Integrating software into PRA: a test-based approach. AB - Probabilistic risk assessment (PRA) is a methodology to assess the probability of failure or success of a system's operation. PRA has been proved to be a systematic, logical, and comprehensive technique for risk assessment. Software plays an increasing role in modern safety critical systems. A significant number of failures can be attributed to software failures. Unfortunately, current probabilistic risk assessment concentrates on representing the behavior of hardware systems, humans, and their contributions (to a limited extent) to risk but neglects the contributions of software due to a lack of understanding of software failure phenomena. It is thus imperative to consider and model the impact of software to reflect the risk in current and future systems. The objective of our research is to develop a methodology to account for the impact of software on system failure that can be used in the classical PRA analysis process. A test-based approach for integrating software into PRA is discussed in this article. This approach includes identification of software functions to be modeled in the PRA, modeling of the software contributions in the ESD, and fault tree. The approach also introduces the concepts of input tree and output tree and proposes a quantification strategy that uses a software safety testing technique. The method is applied to an example system, PACS. PMID- 16268953 TI - Effects of testosterone on lipid peroxidation, lipid profiles and some coagulation parameters in rabbits. AB - The purpose of this study was to investigate the effects of testosterone on some risk factors of atherosclerosis. Twenty-four male New Zealand white rabbits were randomly divided into three groups of eight. The first group was used as control. Second group was injected with 10 mg of testosterone propionate. Third group was castrated bilaterally. At the end of 6 weeks, lipid peroxidation (LPO), lipid profile, fibrinogen (FBN) level and coagulation parameters were evaluated. Testosterone administration decreased the level of high-density lipoprotein cholesterol (HDL-C), while castration increased this level (P < 0.05). Triglyceride (TG) and total cholesterol (TC) levels in the castration group were significantly higher (P < 0.05) than those in the testosterone group. The ratio of HDL-C:low-density lipoprotein cholesterol (LDL-C) decreased, while TC:HDL-C ratio increased (P < 0.05) in the testosterone group. No significant differences were found in the LDL-C and FBN levels among groups. However, there was a tendency for higher FBN level in the testosterone group. Testosterone administration resulted in an increase in the level of LPO (P < 0.05). Clotting time and prothrombin time prolonged in the castration group compared with testosterone group (P < 0.05). As a result, testosterone has exacerbating effect on atherosclerosis risk factors including lipid profile, LPO, FBN and coagulation system. PMID- 16268952 TI - Enhanced non-esterified fatty acids and corticosterone in blood plasma of chickens treated with insulin are significantly depleted by reverse T: minor changes in hypoglycaemia. AB - Previously, it has been observed that dexamethasone or adrenaline-induced hyperlipaemia in blood of chicken was significantly reduced after administration of reverse triiodothyronine (rT3). The present experiment was performed on chicken to determine the altered circulating non-esterified fatty acids (NEFA) induced by physiologically enhanced endogenous corticosterone and catecholamines may also be influenced by rT3. Rise of both hormones were induced by insulin administration. Changes in circulating glucose, corticosterone and catecholamines were additionally measured. Following insulin injection blood glucose fell on the average by 32.7% below control at 2 h of the experiment. Additional treatment with rT3 (rT3 + insulin group) gradually attenuated this decrease and at 4 and 6 h of the experiment it was 17.1% and 12.9% below control, respectively, suggesting on slight inhibition by rT3 of insulin-stimulated glucose utilization. Exposure to insulin significantly increased NEFA levels to about 670% above control group. Additional treatment with rT3 reduced this increase to 309% of control, suggesting inhibition of lipolysis by rT3. Similar alterations were observed in plasma corticosterone levels. Insulin treatment peaked the corticosterone levels maximally by 507.6% above control. Additional treatment with rT3 abolished this rise in the averages to 194.2% above control, possibly by interaction of rT3 with hypothalamo-adrenal axis. Insulin injection increased plasma catecholamines on the average by 21.5% and 53.4% for adrenaline and noradrenaline respectively. Supplementary treatment with rT3 intensified this rise by 55.6% and 71.6% respectively. The obtained results suggest on inhibitory effect of rT3 on hypoglycaemia, hyperlipaemia and plasma corticosterone concentrations in chickens treated with insulin. Contrary to this, rT3 enhanced the rise of plasma catecholamines due to insulin treatment. The obtained data favour the assumption that hypometabolic properties of rT3 depends mainly upon reduced supply of NEFA as a result of restricted lipolysis and to a lesser extent upon the supply of glucose. PMID- 16268954 TI - Effect of ACTH administration on biochemical and immune measures in boars. AB - The effects of stress induced by adrenocorticotropic hormone (ACTH) on biochemical and immune changes in Swedish Landrace boars aged approximately 6-7 months were observed during ACTH administration and for 16 days after the cessation of treatment. Adrenocorticotropic hormone treated animals (n = 7) were given intravenously 10 microg/kg body mass of ACTH for 3 days. Control group of animals (n = 7) received intramuscularly 1 ml of sterile 0.9% saline. Total protein and globulin levels were significantly elevated during the induced stress period and for the next 16 days (P < 0.01 to P < 0.0001, respectively). Also, serum immunoglobulin IgG concentration was significantly elevated during and after ACTH injection (P < 0.001 to P < 0.0001). Adrenocorticotropic hormone treatment had no effect on serum albumin, IgA and IgM concentrations. Glucose concentration was significantly decreased on the second day of ACTH administration (P < 0.001) and on day 9 after treatment (P < 0.001). Calcium level was significantly decreased only for 24 h after the last ACTH dosage (P < 0.01). Also, serum phosphorus level was significantly decreased on the first (P < 0.05) and third (P < 0.001) days of ACTH challenge but remained unaffected after the cessation of ACTH treatment. It is concluded that the administration of ACTH to boars results in immune humoral and biochemical changes during stress and the post-stress period. PMID- 16268955 TI - Immunohistochemical identification of amyloid, using an anti-human serum amyloid P component (SAP) antibody, is possible in ruminants but not in dogs and cats. AB - Amyloidosis represents a heterogenous group of diseases that have in common the deposition of fibrils composed of proteins of beta-pleated sheet structure, a structure which can be specifically identified by histochemistry using the Congo red or similar stains. Amyloid consists primarily of the amyloid fibrils but also of the amyloid P component (AP). This component, which is identical with the serum counterpart (SAP), is found in all types of human amyloid, and immunohistochemical identification of AP has been proposed as an adjunct to the universal, type-independent diagnosis of human amyloidosis. In the present study of animal amyloidosis, we compared the amyloid-specific Congo red stain with an immunohistochemical protocol using an anti-human SAP antibody for the identification of amyloid in formalin fixed tissue samples. The species and types of amyloidoses investigated were: (i) seven cows, one yak (Bos grunniens), and one sheep affected with amyloidosis of presumed AA type, (ii) one dog with a pancreatic endocrine tumour producing amyloid of presumed AIAPP type, (iii) two cats with presumed AIAPP-amyloidosis of the islets of Langerhans, one cat with presumed AA-amyloidosis, and one cat with an amyloid-producing odontogenic tumour. Intense immunostaining co-localized with amyloid, identified by its congophilia and green birefringence, using a protocol without any antigen retrieval in each of the seven cows, the yak and the sheep. The method seemed more sensitive in the ruminants than the Congo red stain, but was unable to detect amyloid in the dog and the cats regardless of the application of various antigen retrieval protocols. However, specific identification of amyloid still rests on the Congo red method or similar histochemical techniques. PMID- 16268956 TI - Immunohistochemical observations on TSH secreting cells in pituitary glands of goat kids with congenital goitre. AB - Pituitary glands of normal-termed stillborn goat kids with congenital goitre and normal-termed stillborn goat kids without congenital goitre were examined macroscopically, histopathologically and immunohistochemically. Thyroid glands of these animals were also examined grossly and microscopically. The pituitary glands of kids with goitre were larger than those of normal kids, and on histopathological examination there was hyperplasia of the acidophil cells in the ventral part of the glands. However, it was impossible to distinguish thyroid stimulating hormone (TSH)-secreting cells from other acidophil cells in sections stained with haematoxylin and eosin (HE). Red granules were observed in the cytoplasm of these hyperplastic cells in periodic acid-Schiff (PAS)-stained sections. Sections were also immunostained with an antibody against TSH using the streptavidin-biotin peroxidase technique. Immunohistochemistry revealed TSH secreting cells to have increased in number in the pituitary glands of kids with congenital goitre because of the extensive proliferation when compared with those of normal kids. The present study indicated that the presence of multiple fetuses (twins or triplets) may be a predisposing factor for congenital goitre. PMID- 16268957 TI - The effect of different isolation procedures on canine leucocyte populations and on lectin-induced lymphocyte proliferation. AB - Proliferation assays performed on peripheral blood mononuclear cells (PBMC) are commonly used in experimental and clinical immunology. A prerequisite for an in vitro assay is the ability to obtain relatively pure populations of mononuclear cells from whole blood, as contaminating polymorphonuclear cells may affect the proliferation of lymphocytes. Purification of canine leucocytes from whole blood is associated with difficulties in obtaining pure lymphocytes in high yields. The aim of this study was to optimize the lymphocyte purification from canine whole blood in terms of total cell recovery and purity, while not influencing the proliferation capacity of the isolated cells. To acquire optimal isolation of canine lymphocytes several density gradient media of different densities and osmolalities were examined. For optimal phagocyte removal, pre-treatment of whole blood with carbonyl iron/arabic gum and/or adherence to fibrinogen pre-coated polystyrene tissue flasks were examined. Lectin-induced proliferation was used as measurement of cell activity of the obtained cell fractions after the different separation procedures. Canine blood pre-treated with carbonyl iron/arabic gum followed by density gradient centrifugation with medium 'G' (density: 1.079 g/cm(3), osmolality: 256 mOsm) and adherence to pre-coated polystyrene tissue flask obtained the best PBMC cultures with a median lymphocyte purity of 88% and a median yield of recovered lymphocytes of 54%. This culture also resulted in the highest proliferation and subsequently the highest stimulation index upon lectin stimulation. PMID- 16268958 TI - Effect of insemination volume on uterine contractions and inflammatory response and on elimination of semen in the mare uterus-scintigraphic and ultrasonographic studies. AB - The effect of artificial insemination (AI) volume on uterine contractility and inflammation and on elimination of semen in the reproductive tract of mares was examined for 4 h after AI using two methods, scintigraphy and ultrasonography. The same doses were used in both methods: 2 and 100 ml of skim milk-extended frozen semen. In the scintigraphic study, the number of reproductively normal mares was four per group and in the ultrasonographic study five per group. For scintigraphy, the semen was radiolabelled with technetium-99m. The static scintigrams were acquired immediately before and 30, 60, 120, 180 and 240 min after AI. The activities in the vagina and uterus were calculated and the values for sperm that had been discharged from the mare were obtained by subtracting the counts for the uterus and vagina from the total radioactivity. The dynamic scintigrams were taken continuously for the first 30 min after AI and in 5-min periods immediately after having acquired the static scintigrams. The uterine contractions were counted. In the ultrasonographic study, the mares were scanned before AI and at 5, 10, 15, 20, 25, 30, 60, 120, 150, 180, and 240 min after AI, for at least 1 min each time. The examinations were videotaped and contractions counted per minute. More contractions were observed with the ultrasonographic method than with the scintigraphic method. No difference was present in the number of contractions between the groups, except in the ultrasonographic study at 4 h, when the mares inseminated with 100 ml showed more contractions than did the mares inseminated with 2 ml. The intraluminal fluid was sampled with a tampon and by uterine lavage 4 h after AI in the ultrasonographic study. The numbers of polymorphonuclear leukocytes and spermatozoa were counted, but the differences between the groups were not significant. Under our experimental conditions and with the number of mares examined, the volume of the AI dose had an insignificant effect on contractility - with the exception at 4 h - and inflammatory reaction and on semen elimination in the uterus. PMID- 16268959 TI - Influence of partus induction on the pH value in the blood of newborn piglets. AB - Partus induction is an important integrative component of reproduction management in the swine industry. In this context, the question is to what extent the application of uterotonic substances may influence the vitality of piglets. The blood pH value is used as a laboratory objective diagnostic parameter of neonatal acidosis. This parameter of the acid-base balance was examined in piglets born spontaneously at the 115th to 116th day of gestation (group 1, n = 57) and after two different partus induction protocols starting at the 114th day of gestation (group 2: combination of 175 microg of cloprostenol and 20 IU of oxytocin, n = 55; group 3: 175 microg of cloprostenol and 35 microg of carbetocin, n = 56). Initial blood samples were taken from each piglet at birth and 2 h later. The results show that the different protocols have no negative influence on the initial pH value and the compensation of neonatal acidosis until the second hour post-natum. PMID- 16268960 TI - Retrospective study on vulvar conformation in relation to endometrial cytology and fertility in thoroughbred mares. AB - Changes in the conformation of the vulva predispose the mare to uterine infection. Vulvoplasty for closure of the upper vulvar lips improves fertility. Not all mares are resutured after parturition, but are resutured after mating. No clinical data have hitherto been published on the reproductive outcome in mares that are resutured or not after parturition. The objectives of the present study were to investigate relationships between vulvar status (not Caslick-operated, group A; previously Caslick-operated and/or not resutured after the last parturition, groups B and C respectively) and endometrial cytology, conception rate and live foal rate for different age groups and categories of Thoroughbred mares. A careful examination of the vulvar area, and endometrial sampling for bacteriology and cytology was performed. The mares were naturally mated once during oestrus (never foal heat) by one of two stallions. Mares belonging to vulvar status group A had the highest conception rate and live foal rate, whereas the lowest conception rate and live foal rate occurred in mares with vulvar status group C. A multifactorial analysis revealed that the vulvar status at arrival had a significant effect on pregnancy rate and live foal rate, but not on fetal loss. Stallion, year and age group had no significant effect on any of the three variables analysed. In conclusion, optimizing managerial factors such as resuturing Caslick-operated mares immediately after parturition to minimize growth of micro-organisms with a subsequent endometritis will contribute to their conception and that they carry a pregnancy to term. PMID- 16268961 TI - Interdigital involvement in a case of primary cutaneous canine histoplasmosis in Japan. AB - A 5-year-old male Siberian husky bred outdoor in Tokyo had a swollen paw with interdigital granulomatous lesions in the left hindlimb. The dog had no apparent pulmonary or gastrointestinal involvement. Histopathological analysis of the skin lesions demonstrated yeast-like organisms predominantly within macrophages. Sequence analysis of fungal ribosome RNA gene isolated from a paraffin sample revealed a 100% homology with the teleomorph of Histoplasma capsulatum. The present case may support the concept of primary cutaneous canine histoplasmosis as an endemic phenotype recognized in Japan. PMID- 16268962 TI - Osseous lesion of the calcaneus following the use of shock wave therapy in a horse. AB - An 8-year-old Dutch warmblood gelding was presented with a mechanical lameness (2/10) because of the presence of a soft tissue injury on the top of the right tuber calcanei. Plain radiographs of the tarsus demonstrated the presence of soft tissue swelling caudal to the right tuber calcanei, without osseous involvement, and ultrasonography revealed excessive scar tissue within and around the superficial digital flexor tendon. Extra-corporeal shock wave therapy was applied on the right hock to decrease the amount of scar tissue. One month after the therapy the lameness was greater (3/10) and a marginal increase in the size of the swelling was found. Periosteal new bone formation associated with an ill defined radiolucent area and two bony fragments were detected radiographically at the caudo-proximal aspect of the right tuber calcanei. A blister containing oil of croton, camphor, pine and thyme, turpentine and cantharides was applied on the right calcaneus. Twenty days after blister application, the size of the swelling had been reduced by 50% and the degree of lameness had also been decreased (1/10). On clinical re-evaluation 6 months after treatment, the degree of lameness was stable (1/10) and flexion test of the limb was negative. PMID- 16268963 TI - Review article: the treatment of non-alcoholic steatohepatitis with thiazolidinediones. AB - It is generally accepted that non-alcoholic fatty liver disease will be the most frequent liver disease in the near future and that the management of patients with non-alcoholic fatty liver disease will be a challenge for hepatologists in the next decades. Non-alcoholic fatty liver disease is considered the hepatic manifestation of the metabolic syndrome, in which insulin resistance plays a crucial role. Although steatosis will often not progress to severe liver disease, in some patients, it results in cirrhosis and even hepatocellular carcinoma. Therefore, it is important to identify those patients at risk for developing fibrosis. Age, diabetes, obesity and hypertriglyceridaemia are independent risk factors for fibrosis in patients with elevated serum alanine aminotransferase levels and steatosis on ultrasound. The presence of multiple metabolic disorders increases the risk. Apart from diet, exercise and correction of underlying metabolic abnormalities, no specific treatment is available at the moment. Theoretically, thiazolidinediones are an attractive way to treat non-alcoholic fatty liver disease, because they improve insulin resistance. Some preliminary studies with thiazolidinediones were encouraging, as steatosis, inflammation and fibrosis improved in a substantial number of patients. Although no serious side effects occurred in the pilot studies, we should look vigilantly for hepatotoxicity, as the first generation thiazolidinediones proved to be toxic for the liver. PMID- 16268964 TI - Review article: how and when to use ciclosporin in ulcerative colitis. AB - Although colectomy for ulcerative colitis is curative, long-term quality of life is reduced. Intravenous ciclosporin 4 mg/kg/day has significant toxicity. There is now evidence that low-dose ciclosporin (2 mg/kg daily by intravenous infusion, or 5-6 mg/kg daily in a twice daily oral dosage) has an acceptable safety profile, even when used in combination with corticosteroids. Drug dosage should be adjusted to the levels of 150-250 ng/mL initially (random levels during intravenous infusion, or trough levels for oral use). Ciclosporin should be considered not only in those who have failed 7 days of corticosteroids, but also in fulminant colitis at day 3, if not responding to corticosteroids. The drug should be avoided in frail or elderly patients with significant comorbidity, and also where colectomy is likely to be necessary in the short to medium term. Ciclosporin should not be continued for more than 7 days, unless there is a definite response. A 70-80% initial response is likely, and responders are discharged on oral ciclosporin, adding thiopurines and tailing prednisolone rapidly. The drug should be continued for 3 months. The likelihood of avoiding colectomy over 2-3 years is 40-50%. More studies are needed to evaluate the use of oral ciclosporin in corticosteroid-refractory colitis in out-patients, and to assess whether monotherapy (without corticosteroids) is significantly safer, without loss of efficacy. PMID- 16268965 TI - Systematic review: acute colonic pseudo-obstruction. AB - Acute colonic pseudo-obstruction is the clinical syndrome of acute large bowel dilatation without mechanical obstruction that is an important cause of morbidity and mortality. Acute colonic pseudo-obstruction occurs in hospitalized or institutionalized patients with serious underlying medical and surgical conditions. The pathogenesis of acute colonic pseudo-obstruction is not completely understood but likely results from an imbalance in the autonomic regulation of colonic motor function. Metabolic or pharmacological factors, as well as spinal or retroperitoneal trauma, may alter the autonomic regulation of colonic function, leading to excessive parasympathetic suppression or sympathetic stimulation. This imbalance results in colonic atony and dilatation. Early recognition and appropriate management are critical to minimizing morbidity and mortality. The mortality rate is estimated at 40% when ischaemia or perforation occurs. The best-studied treatment of acute colonic pseudo-obstruction is intravenous neostigmine, which leads to prompt colon decompression in the majority of patients after a single infusion. In patients failing or having contraindications to neostigmine, colonoscopic decompression is the active intervention of choice. Surgery is reserved for those with peritonitis or perforation. PMID- 16268966 TI - Melatonin improves bowel symptoms in female patients with irritable bowel syndrome: a double-blind placebo-controlled study. AB - BACKGROUND: Melatonin is involved in the regulation of gastrointestinal motility and sensation. AIM: To determine the potential therapeutic effects of melatonin in irritable bowel syndrome (IBS). METHOD: Seventeen female patients satisfying the Rome II criteria for IBS were randomized to receive either melatonin 3 mg nocte or identically appearing placebo 1 nocte for 8 weeks, followed by a 4-week washout period and placebo or melatonin in the reverse order for another 8 weeks. Three validated questionnaires - the GI symptom, the sleep questionnaires and the Hospital Anxiety and Depression Scale - were used to assess symptom severity and to compute the IBS, sleep and anxiety/depression scores, respectively. RESULTS: Improvements in mean IBS scores were significantly greater after treatment with melatonin (3.9 +/- 2.6) than with placebo (1.3 +/- 4.0, P = 0.037). Percent response rate, defined as percentage of subjects achieving mild-to-excellent improvement in IBS symptoms, was also greater in the melatonin-treated arm (88% vs. 47%, P = 0.04). The changes in mean sleep, anxiety, and depression scores were similar with either melatonin or placebo treatment. CONCLUSIONS: Melatonin is a promising therapeutic agent for IBS. Its therapeutic effect is independent of its effects on sleep, anxiety or depression. PMID- 16268967 TI - Prevalence and demographics of irritable bowel syndrome: results from a large web based survey. AB - BACKGROUND: Irritable bowel syndrome is a common gastrointestinal disorder, and its prevalence and demographics have been evaluated by different methodologies with varying results. AIM: To evaluate irritable bowel syndrome demographic and prevalence characteristics utilizing a web-enabled panel. METHODS: From an existing 150 000-member panel, 31 829 individuals were randomly selected and sent screening questionnaires to evaluate irritable bowel syndrome symptoms. Individuals who agreed to participate and completed the screening questionnaire received a second questionnaire related to a diagnosis of irritable bowel syndrome, a more detailed symptom description, and additional burden of illness data. RESULTS: Irritable bowel syndrome prevalence was 7%. Prevalence was higher in women vs. men, unmarried individuals vs. married individuals and unemployed individuals vs. employed individuals. Of those completing the second questionnaire, 51% had seen their physicians for irritable bowel syndrome symptoms in the past year and most had an episode within the past 3 months. During the past year, approximately half of the participants had used a prescription medication, and over 90% had used an over-the-counter medication for irritable bowel syndrome. Participants with irritable bowel syndrome demonstrated quality-of-life reductions relative to norms of the United States population. CONCLUSIONS: Web-enabled data collection represents a novel tool for rapidly surveying a large population of individuals with irritable bowel syndrome symptoms. PMID- 16268968 TI - Hepatic steatosis in chronic hepatitis C: impact on response to anti-viral treatment with peg-interferon and ribavirin. AB - BACKGROUND: There is increasing evidence that hepatic steatosis contributes to the progression of liver fibrosis, whereas its impact on the efficacy of anti viral treatment is still under investigation. AIM: To evaluate the effect of steatosis on the outcome of combined anti-viral treatment. METHODS: We studied 102 consecutive naive patients with chronic hepatitis C receiving combined anti viral therapy (peg-interferon alpha-2b and ribavirin). RESULTS: Fifty (49%) of 102 patients had evidence of hepatic steatosis (29 grade 1, 16 grade 2 and 5 grade 3). Sustained virological response was similar in patients with and without steatosis (58% vs. 56%); moreover, the grade of steatosis did not affect the rate of sustained virological response (grade 1: 58%, grade 2: 56% and grade 3: 60%). Patients with steatosis had significantly higher serum levels of aspartate transaminase, alanine transaminase and gamma-glutamyltransferase (P = 0.007, 0.004 and 0.03, respectively), higher histological activity (P = 0.03), more advanced stage of fibrosis (P = 0.0394) and more often hepatitis C virus genotype 3 (P = 0.04). CONCLUSIONS: Our findings suggest that hepatic steatosis in chronic hepatitis C, irrespective of its grade, is not a negative prognostic factor of response to combined anti-viral therapy, even when the histological and biochemical profile of the disease is more aggressive. PMID- 16268969 TI - Anti-cyclic citrullinated peptide antibodies in type 1 autoimmune hepatitis. AB - BACKGROUND: Besides the autoantibodies included in the diagnostic criteria of type 1 autoimmune hepatitis, many other autoantibodies have been described in this condition. Recently, antibodies against cyclic citrullinated peptide have been validated as specific diagnostic and prognostic markers of rheumatoid arthritis. AIM: To assess whether these antibodies are part of the autoantibody repertoire of type 1 autoimmune hepatitis and correlate with rheumatological manifestations. METHODS: Antibodies against cyclic citrullinated peptide were tested by a commercially available enzyme-linked immunosorbent assay. RESULTS: The antibodies were found in 12 of 133 (9%) type 1 autoimmune hepatitis, two of 49 (4%) with primary biliary cirrhosis, one of 80 (1%) with hepatitis C virus related chronic liver disease and 53 of 89 (60%) with rheumatoid arthritis serum samples. High titres were found only in rheumatoid arthritis and type 1 autoimmune hepatitis. No clinical (in particular rheumatological manifestations), biochemical or immunoserological differences were detectable between antibodies against cyclic citrullinated peptide positive and negative type 1 autoimmune hepatitis sera, with the exception of rheumatoid factor, always negative in the positive ones. CONCLUSIONS: Antibodies against cyclic citrullinated peptide can be detected in a subgroup of patients with type 1 autoimmune hepatitis. They might be part of the wide range of autoantibody production characteristic of this condition and/or, less probably, be predictive of future rheumatoid arthritis development. PMID- 16268970 TI - Capsule endoscopy - comparison of two strategies of bowel preparation. AB - BACKGROUND: Capsule endoscopy can identify lesions of the small bowel that cannot be detected by other techniques. In addition to patient safety, quality of care and cost-efficiency, good preparation is an important factor for good visualization. AIM: To evaluate the efficacy of oral sodium phosphate preparation. METHODS: Forty-six consecutive patients scheduled for capsule endoscopy in two medical centres. The patients treated in Dallas were prepared by 12 h fasting (group A), and those treated in Israel were also asked to drink 45 mL of sodium phosphate with water (group B). An experienced endoscopist, blinded to the method used, graded the quality of preparation according to visual capability, and to relative durations of each grade. RESULTS: The quality of the preparation was poor in 35% of group A compared with 4% of group B (P = 0.023). The mean duration of good preparation with excellent visualization was 122 +/- 110 min in group A and 180 +/- 96 min in group B (P = 0.006). Preparation with sodium phosphate and lower patient weight were significant predictive factors for good visualization. CONCLUSIONS: Bowel preparation offers better visualization than overnight fasting alone and is associated with fewer disturbances by intraluminal turbid fluid. PMID- 16268971 TI - Efficacy of depot long-acting release octreotide therapy in severe dumping syndrome. AB - BACKGROUND: Dumping syndrome is a serious complication occurring in 10% of patients after gastric surgery. Dumping symptoms are effectively reduced by subcutaneous application of the somatostatin analogue octreotide, but side effects limit its use. AIM: To evaluate the efficacy of depot long-acting release octreotide (Sandostatin-LAR) vs. octreotide subcutaneous on dumping symptoms, quality of life and side-effects. METHODS: Twelve patients (five females, age 58 +/- 3 years) with severe dumping symptoms, requiring daily use of octreotide subcutaneous, were included in an open study and changed from octreotide subcutaneous after a 2 weeks washout to Sandostatin-LAR 10 mg i.m., every 4 weeks for 6 months. Symptoms (diary), body weight, fat excretion, food intake and Gastrointestinal Specific Quality of Life Index were evaluated. RESULTS: Gastrointestinal Specific Quality of Life Index increased significantly (P < 0.05) during Sandostatin-LAR treatment (88 +/- 4) compared with octreotide (74 +/ 4) and washout (75 +/- 6). During Sandostatin-LAR treatment, abdominal symptom score was lower compared with octreotide and washout, but not significantly. During Sandostatin-LAR treatment, body weight increased (66 +/- 4 to 70 +/- 3 kg; P = 0.19). CONCLUSIONS: Sandostatin-LAR is at least as effective as octreotide subcutaneous in suppressing symptoms in patients with severe dumping syndrome and is more effective than octreotide subcutaneous in increasing body weight and quality of life. PMID- 16268972 TI - Intestinal fluid volumes and transit of dosage forms as assessed by magnetic resonance imaging. AB - AIM: The gastrointestinal transit of sequentially administered capsules was investigated in relation to the availability of fluid along the intestinal lumen by magnetic resonance imaging. METHODS: Water-sensitive magnetic resonance imaging was performed on 12 healthy subjects during fasting and 1 h after a meal. Specifiable non-disintegrating capsules were administered at 7, 4 and 1 h prior to imaging. RESULTS: While food intake reduced the mean fluid volumes in the small intestine (105 +/- 72 mL vs. 54 +/- 41 mL, P < 0.01) it had no significant effect on the mean fluid volumes in the colon (13 +/- 12 mL vs. 18 +/- 26 mL). The mean number of separated fluid pockets increased in both organs after meal (small intestine: 4 vs. 6, P < 0.05; large intestine: 4 vs. 6, P < 0.05). The distribution of capsules between the small and large intestine was strongly influenced by food (colon: 3 vs. 17 capsules, P < 0.01). CONCLUSIONS: The results show that fluid is not homogeneously distributed along the gut, which likely contributes to the individual variability of drug absorption. Furthermore, transport of fluid and solids through the ileocaecal valve is obviously initiated by a meal-induced gastro-ileocaecal reflex. PMID- 16268973 TI - Gabapentin reduces rectal mechanosensitivity and increases rectal compliance in patients with diarrhoea-predominant irritable bowel syndrome. AB - BACKGROUND: Gabapentin has been shown to reduce elements of central sensitization in human experimental hyperalgesia. It remains uninvestigated whether gabapentin has beneficial effects for irritable bowel syndrome associated with visceral hypersensitivity. AIMS: To evaluate the effects of gabapentin on sensory and motor function of the rectum in patients with diarrhoea-predominant irritable bowel syndrome. METHODS: Forty patients with diarrhoea-predominant irritable bowel syndrome completed this randomized, double-blind, placebo-controlled, parallel-grouped study. All patients received a barostat study and were subsequently randomized for 5-day treatment with gabapentin 300 mg/day and then 600 mg/day or placebo. On day 6, after subjects had their morning dose, the barostat experiment was repeated. RESULTS: The threshold pressures for bloating, discomfort and pain significantly increased after gabapentin, but not after placebo. Significant increase in the pressure and corresponding wall tension inducing discomfort or pain were observed in the gabapentin group, but not in the placebo group. Rectal compliance significantly increased after gabapentin, but not after placebo. The postprandial increase of rectal tone was not affected by gabapentin. CONCLUSION: Our results show that gabapentin reduces rectal sensory thresholds through attenuating rectal sensitivity to distension and enhancing rectal compliance in diarrhoea-predominant irritable bowel syndrome patients. The clinical efficacy of this drug in irritable bowel syndrome patients warrants investigation. PMID- 16268974 TI - Low rectal volumes in patients suffering from fecal incontinence: what does it mean? AB - BACKGROUND: Rectal perception and adaptation to distension are widely heterogeneous in subjects with faecal incontinence. AIM: To quantify rectal physiology in patients with incontinence and low maximum rectal volume, according to AGA guidelines on anorectal testing techniques. PATIENTS AND METHODS: 148 patients (12 men, 136 female) with incontinence to liquid and/or solid stools were investigated. Distending isobaric procedures were carried out using an electronic barostat in order to analyse perception and adaptation of the rectum. RESULTS: Pain during isovolumic rectal distension at a level of 100 mL or less was experienced in 21 subjects (14.2%). As defined by isobaric distensions, incontinent patients with low MTV had more frequently a hypocompliant rectum (62%) when compared with those with higher MTV (31%, P = 0.046). Perception scores tended to be higher at each step of distending rectal pressure: incontinent patients with low MTV had more frequently a hypersensitive rectum (48%) when compared with those with normal or high MTV (24%, P = 0.035). Only four of 21 incontinent subjects with low MTV had an isolated hypersensitive rectum. CONCLUSION: Both sensitivity and compliance are altered in patients with low MTV. A more extensive study of the role of sensory and compliance aspects of subjects with incontinence is warranted. PMID- 16268975 TI - Outlet type constipation in Parkinson's disease: results of botulinum toxin treatment. AB - BACKGROUND: Constipation is one of the most common autonomic dysfunctions observed in Parkinson's disease. AIM: To investigate the efficacy of injections of botulinum toxin in improving rectal emptying in these patients. METHODS: Eighteen Parkinson's disease patients with outlet constipation were included in the study. The patients were treated with type A botulinum toxin, injected into two sites on either side of the puborectalis muscle under ultrasonographic guidance. RESULTS: Symptomatic improvement was noted in 10 patients, at 2 months evaluation. In these subjects, anorectal manometry demonstrated decreased tone during straining from 96.2 +/- 17.1 to 45.9 +/- 16.2 mmHg at 1 month evaluation (P = 0.00001) and to 56.1 +/- 10.7 mmHg at 2 months (P = 0.00001). Pressure during straining was lower than resting anal pressure at the same times in all patients. Defecography after the treatment showed improvement in anorectal angle during straining, which increased from 99.1 +/- 8.4 degrees to 121.7 +/- 12.7 degrees (P = 0.00001) at 2 months. CONCLUSIONS: Botulinum toxin injections may be a useful treatment for Parkinson's disease patients affected by outlet obstruction constipation. The treatment is safe and simple. However, because the effects of the toxin wear off within 3 months of administration, repeated injections could be necessary to maintain the clinical improvement. PMID- 16268976 TI - The association of body mass index with Barrett's oesophagus. AB - BACKGROUND: Obesity has been linked to gastro-oesophageal reflux disease symptoms and oesophageal adenocarcinoma; however, there is no published evidence for an association with Barrett's oesophagus. AIM: To investigate the association between obesity and Barrett's oesophagus. METHODS: We conducted a retrospective cross-sectional study of patients who underwent upper endoscopy at the Southern Arizona Veteran's Affairs Healthcare System between 1998 and 2004. We examined male patients without malignancy, with available information on weight and height. Based on endoscopic and histological findings, patients were classified as cases with Barrett's oesophagus or non-cases without Barrett's oesophagus. Multivariable logistic regression analysis was conducted to examine the association of body mass index and obesity with Barrett's oesophagus and Barrett's oesophagus length while adjusting for age and race. RESULTS: There were 65 cases with Barrett's oesophagus and 385 non-cases without Barrett's oesophagus. The mean body mass index was significantly higher in cases than in non-cases (29.8 vs. 28.0, P = 0.03). Cases had significantly greater mean weight than controls (206 lb vs. 190,P = 0.005). The proportions of cases with body mass index 25-30 and body mass index > or =30 were greater than those in non-cases (44.6% vs. 37.7%) and (40.0% vs. 33.5%), respectively (P = 0.08). In the multivariable logistic regression model adjusting for race and age, when compared with body mass index < 25, the odds ratio was 2.43 (95% confidence interval: 1.12 5.31) for body mass index 25-30 and 2.46 (1.11-5.44) for body mass index > or =30. When examined as a continuous variable the adjusted odd ratio for each five point increase in body mass index was 1.35 (95% confidence interval: 1.06-1.71, P = 0.01). The association between weight and Barrett's oesophagus was also statistically significant (adjusted odd ratio for each 10 pound increase = 1.10, 1.03-1.17, P =0.002). Among the 65 cases of Barrett's oesophagus, there was no correlation between the length of Barrett's oesophagus at the time of diagnosis and the body mass index (correlation coefficient = 0.03, P = 0.79). CONCLUSION: This retrospective cross-sectional study in male veterans shows that overweight is associated with a two-and-half-fold increased risk of Barrett's oesophagus. Larger studies of the underlying mechanism are warranted to better understand how and why obese patients are at greater risk for Barrett's oesophagus. PMID- 16268977 TI - Normal values and day-to-day variability of 24-h ambulatory oesophageal impedance pH monitoring in a Belgian-French cohort of healthy subjects. AB - BACKGROUND: Impedance-pH monitoring is the most sensitive method for detection and characterization of gastro-oesophageal reflux episodes. Normal values from European subjects are lacking. AIM: To build a database of gastro-oesophageal reflux patterns from French and Belgian healthy subjects. METHODS: Seventy-two healthy subjects (35 men, mean age 35 years, 18-72) underwent 24-h ambulatory impedance-pH studies. Gastro-oesophageal reflux episodes were detected using impedance and characterized by pH as acid, weakly acidic, or weakly alkaline. Analysis was performed visually and effects of age, gender and intra-individual reproducibility were evaluated. RESULTS: The total number of gastro-oesophageal reflux episodes was 44 (25,58,75) of which 59% were acid, 28% were weakly acidic and 10% weakly alkaline. Half of gastro-oesophageal reflux episodes were mixed (liquid/gas) and 22% reached 15 cm above the lower oesophageal sphincter. The bolus clearance time was 11 s while acid was chemically cleared in 34 s. Male gender was associated with increased number and proximal extent of total and acid gastro-oesophageal reflux. Repeated studies in 27 subjects showed good reproducibility for number, acidity and air-liquid composition of reflux (Kendall's W-values = 0.72-0.85). CONCLUSIONS: This study demonstrates good reproducibility of 24-h ambulatory impedance-pH studies and provides values of reflux patterns in healthy subjects for comparisons with European gastro oesophageal reflux disease patients. PMID- 16268978 TI - Risk factors for non-cardiac chest pain in the community. AB - BACKGROUND: Non-cardiac chest pain is a common and expensive condition. Risk factors for non-cardiac chest pain are poorly understood and lack description in the USA community. AIM: To explore risk factors and health-seeking behaviour in non-cardiac chest pain in a USA community. METHODS: Olmsted County, MN subjects who reported at least monthly or severe chest pain in response to a validated questionnaire were compared with controls. All the Mayo Clinic medical records were reviewed; those with cardiac disease or relevant organic conditions were excluded. RESULTS: Of 2118 eligible subjects, 1524 (72%) returned a questionnaire, 389 subjects (26%) reported any chest pain and 186 (12.2%) at least monthly or severe chest pain. Of these, 62 were excluded for a cardiac diagnosis or another organic cause. Thus, 124 subjects (9.1%, 95% CI: 7.6-10.8%) reported at least monthly or severe non-cardiac chest pain, of which 65 (52.4%) had frequent reflux symptoms. Independent risk factors for non-cardiac chest pain were obesity (OR 3.0, 95% CI: 1.64-5.50), family history of reflux (OR 2.8, 95% CI: 1.73-4.32), previous cigarette use (OR 2.0, 95% CI: 1.27-3.18), aspirin use (OR 1.5, 95% CI: 1.00-2.31) and use of antiarthritis medicines (OR 2.0, 95% CI: 1.27-3.16). Compared with subjects with non-cardiac chest pain and associated gastro-oesophageal reflux symptoms, subjects with non-cardiac chest pain without associated gastro-oesophageal reflux symptoms were less likely to have a family history of reflux, more likely to be younger, and less likely to be obese. Compared with controls, subjects with non-cardiac chest pain without gastro oesophageal reflux symptoms were younger (OR 0.97, 95% CI: 0.95-0.99), reported higher somatic symptom scores (OR 1.1, 95% CI: 1.08-1.73) and were more likely to be obese (OR 2.6, 95% CI: 1.15-5.93). CONCLUSIONS: Recurrent or severe non cardiac chest pain is common in the community. Half of the people with significant non-cardiac chest pain have frequent reflux symptoms, thus, risk factors for non-cardiac chest pain are similar to risk factors for gastro oesophageal reflux disease. People with non-cardiac chest pain without reflux symptoms have a slightly different risk factor profile. PMID- 16268979 TI - Poor metabolizer genotype status of CYP2C19 is a risk factor for developing gastric cancer in Japanese patients with Helicobacter pylori infection. AB - BACKGROUND: Cytochrome P450 2C19 (CYP2C19) polymorphism has been associated with the development of lung, liver or oesophageal cancer by detoxification of carcinogen(s) or activation of procarcinogen(s). AIM: To clarify the association between CYP2C19 polymorphisms and gastric cancer development in Japanese. Methods : We determined CYP2C19 genotypes (CYP2C19*1, *2 and *3) in 111 Helicobacter pylori-positive patients with gastric cancer and 315 H. pylori-positive controls without gastric cancer consisting of patients with gastritis only or peptic ulcer. Frequencies of CYP2C19 genotypes and serum pepsinogen I and II levels, a biomarker of gastric atrophy, in the gastric cancers and controls were compared. RESULTS: Frequencies of homozygous extensive metabolizers, heterozygous extensive metabolizers and poor metabolizers were 31.5%, 42.3% and 26.2% in the gastric cancers and 38.1%, 47.0% and 14.9% in the controls, respectively (P = 0.046). Poor metabolizers were associated with an increased risk for developing gastric cancer with the age- and sex-adjusted odds ratio (OR) of 1.975 [95% confidence interval (CI): 1.068-3.649], especially for diffuse type (OR: 3.385, CI: 1.187 9.648). There is no significant association between CYP2C19 genotypes and serum pepsinogen I level or pepsinogen I/II ratios, although serum pepsinogen I level in gastric cancers were significantly decreased. CONCLUSIONS: In H. pylori positive Japanese, poor metabolizers of CYP2C19 appear to be at an increased risk for developing gastric cancer, especially diffuse type, and may require an intensive follow-up for scrutinizing possible gastric cancer development. PMID- 16268980 TI - Helicobacter pylori first-line treatment and rescue options in patients allergic to penicillin. AB - BACKGROUND: Helicobacter pylori eradication is a challenge in patients allergic to penicillin, especially those who have failed a first-eradication trial. AIM: To assess the efficacy and tolerability of H. pylori first-line treatment and rescue options in patients allergic to penicillin. METHODS: Prospective single centre study including 40 consecutive treatments administered to patients allergic to penicillin. Therapy regimens: First-line (12 patients) omeprazole, clarithromycin and metronidazole for 7 days; second-line (17 patients) ranitidine bismuth citrate, tetracycline and metronidazole for 7 days; third-line (nine patients) rifabutin, clarithromycin and omeprazole for 10 days; and fourth-line (two patients) levofloxacin, clarithromycin and omeprazole for 10 days. OUTCOME VARIABLE: a negative (13)C-urea breath test 8 weeks after completion of treatment. RESULTS: Per-protocol/intention-to-treat eradication rates were: first line (64/58%); second-line (ranitidine bismuth citrate; 53/47%); third-line (rifabutin; 17/11%) and fourth-line regimen (levofloxacin; 100/100%). Compliance with treatment was generally good, except with the rifabutin-based regimen, which presented adverse effects in 89% of the patients, including four cases of myelotoxicity. CONCLUSIONS: H. pylori-infected patients who are allergic to penicillin may be treated with a first-line treatment combining a proton-pump inhibitor, clarithromycin and metronidazole. Rescue options may include a regimen with ranitidine bismuth citrate, tetracycline and metronidazole. A levofloxacin based rescue regimen (with proton-pump inhibitor and clarithromycin) may also represent an alternative, even when two or more consecutive eradication treatments have previously failed. However, rifabutin + clarithromycin + proton pump inhibitor regimen is ineffective and poorly tolerated. PMID- 16268981 TI - Cardiorespiratory effects of warm water immersion in elderly patients with chronic heart failure. AB - BACKGROUND: Hydrotherapy might be included in the rehabilitation of patients with chronic heart failure (CHF), but little is known about the acute cardiorespiratory reaction in warm water. The aim of this study was to assess the acute cardiorespiratory effect of immersion in warm water, in a clinical setting, in elderly patients with CHF compared with healthy age and sex matched persons. METHODS: Twelve patients (three females) with CHF, NYHA II-III, age 64 +/- 6 years, and 12 healthy subjects were studied. Cardiorespiratory changes, on land and in a temperature-controlled swimming pool (33-34 degrees C) were assessed during rest and exercise, in a sitting position, using continuous gas analyses. RESULTS: There were no significant differences, land versus water, in carbon dioxide production, total ventilation, respiratory frequency, respiratory exchange ratio, heart rate or blood pressure in either of the groups. A significant difference was found in oxygen uptake, at rest, land versus water in patients with CHF in comparison with healthy subjects (-0.2 +/- 0.4 versus +0.3 +/- 0.6 ml kg(-1) min(-1), P < 0.01). Oxygen kinetics (tau) increased significantly (P = 0.01) in both groups during exercise in water. CONCLUSION: Hydrotherapy was well tolerated and the vast majority of the cardiorespiratory responses, during warm water immersion in a clinical setting, are similar in patients with CHF compared with healthy subjects. However, further larger studies, are needed to better understand the physiological reactions during hydrotherapy. PMID- 16268982 TI - Normal values of pulmonary capillary wedge pressure and the blood pressure response to the Valsalva manoeuvre in healthy elderly subjects. AB - The blood pressure response to the Valsalva manoeuvre is related to pulmonary capillary wedge pressure (PCWP) and can be used to diagnose heart failure. However, this has never been studied specifically in the elderly, in whom the prevalence of heart failure is highest. Furthermore, normal values of the Valsalva manoeuvre are lacking. We aimed to obtain normal values of PCWP and the blood pressure response to the Valsalva manoeuvre in elderly subjects. Therefore, 28 healthy subjects, aged 70 +/- 4 years, performed Valsalva manoeuvres before and after anti-G garment inflation, which was used for temporary increase of PCWP. Before inflation, PCWP was 9.8 +/- 1.9 mmHg in supine and 8.9 +/- 2.1 in semi-recumbent position. From the blood pressure response, measured with Finapres, the systolic blood pressure ratio (SBPR), pulse pressure ratio (PPR), stroke volume ratio (SVR) and heart rate ratio (HRR) were calculated. In supine position, SBPR was 0.76 +/- 0.11, PPR 0.51 +/- 0.16, SVR 0.42 +/- 0.11, and HRR 1.17 +/- 0.12. Semi-recumbently, SBPR was 0.74 +/- 0.10, PPR 0.46 +/- 0.14, SVR 0.41 +/- 0.10, and HRR 1.24 +/- 0.23. After inflation of the anti-G garment, the areas under the Receiver Operator Characteristics curves of SBPR, PPR and SVR for elevated (> or = 15 mmHg) PCWP were >0.85 in supine position. In conclusion, this is the first study to obtain normal values of the blood pressure response to the Valsalva manoeuvre and PCWP in healthy elderly subjects, which is essential for the interpretation of patient data. The Valsalva manoeuvre showed significant discriminatory power in the detection of elevated PCWP, which underscores its potential in the non-invasive diagnosis of heart failure. PMID- 16268983 TI - The effects of left ventricular hypertrophy on the respiratory changes in transmitral Doppler flow patterns of hypertension patients. AB - BACKGROUND: Left ventricular early diastolic fillings can be reduced by inspiration. However, the effects of left ventricular hypertrophy on such changes have not been studied before. This study was undertaken to investigate whether respiratory changes in transmitral Doppler flow were affected by left ventricular hypertrophy in hypertension patients. METHODS: Eighty-three patients (mean age 46 +/- 8 years, 49 males) with untreated essential hypertension were included in this study. Transmitral Doppler flow velocity was measured both at end-expiration and end-inspiration. Left ventricular mass was measured by M-mode echocardiography. We divided patients into two groups based on the presence of left ventricular hypertrophy or not. RESULTS: Twenty-one patients were diagnosed to have left ventricular hypertrophy. In patients without left ventricular hypertrophy, the peak early filling velocity decreased significantly (from 74 +/- 15 to 71 +/- 18 cm s(-1), P = 0.003), the peak atrial velocity increased significantly (from 65 +/- 17 to 74 +/- 15 cm s(-1), P < 0.001) and the early filling to atrial velocity ratio decreased significantly (from 1.2 +/- 0.3 to 1.1 +/- 0.3, P < 0.001) from end-expiration to end-inspiration. In patients with left ventricular hypertrophy, the parameters of transmitral Doppler flow pattern did not change during respiration. CONCLUSION: Respiratory changes in the transmitral Doppler flow velocity are blunted by left ventricular hypertrophy in hypertension patients. This phenomenon is probably contributed by the increased left ventricular wall stiffness in the left ventricular hypertrophy. PMID- 16268984 TI - Physiological determinants of the variation in left ventricular mass from early adolescence to late adulthood in healthy subjects. AB - BACKGROUND: The physiological determinants of left ventricular mass (LVM) measured by cardiac magnetic resonance (CMR) imaging are not well defined as prior investigators have studied either adults or adolescents in isolation or have not strictly excluded hypertension or accounted for the effects of exercise habits, haemodynamic, demographic, or body shape characteristics. METHODS: A total of 102 healthy volunteers (12-81 years, 53 males) underwent CMR. All parameters [unstandardized and adjusted for body surface area (BSA)] were analysed according to gender and by adolescence versus adulthood (adolescents <20 years, adults > or = 20 years). The influence of haemodynamic factors, exercise, and demographic factors on LVM were determined with multivariate linear regression. RESULTS: LVM rose during adolescence and declined in adulthood. LVM and LVMBSA were higher in males both in adults (LVM: 188 +/- 22 g versus 139 +/- 21 g, P < 0.001; LVMBSA: 94 +/- 11 g m(-2) versus 80 +/- 11 g m(-2), P < 0.001) and in adolescents when adjusted for BSA (LVM: 128 +/- 29 g versus 107 +/- 20 g, P = 0.063; LVMBSA: 82 +/- 8 g m(-2) versus 71 +/- 10 g m(-2), P = 0.025). In adults, systolic blood pressure (SBP) and self-reported physical activity increased while meridional and circumferential wall stress were constant with age. Multivariate regression analysis revealed age, gender, and BSA as the major determinants of LVM (global R2 = 0.69). CONCLUSIONS: Normal LVM shows variation over a broad age range in both genders with a rise in adolescence and subsequent decline with increasing age in adulthood despite an increase in SBP and physical activity. BSA, age, and gender were found to be major contributors to the variation in LVM in healthy adults, while haemodynamic factors, exercise, and wall stress were not. PMID- 16268985 TI - Modulation of the inflammatory response to histamine by terbutaline and sodium nitroprusside in guinea-pig skin. AB - We measured the microvascular response (vasodilatation and plasma exudation) to skin prick provocations with histamine, terbutaline, sodium nitroprusside (SNP) and the combinations of terbutaline and histamine as well as SNP and histamine in guinea-pig skin. The response was measured by external detection of beta radiation from transferrin labelled with (113m)In. Histamine induced a moderate microvascular response. Terbutaline alone induced a smaller response, probably reflecting vasodilatation. When added to histamine, terbutaline significantly reduced the microvascular response to histamine. The response to histamine, SNP and the combination of histamine and SNP were all similar. We conclude that the anti-inflammatory effect of terbutaline can be readily measured with this technique. We found no indication of a pro-inflammatory effect of SNP when combined with histamine. Rather, the lack of additive effect may suggest an anti inflammatory effect of SNP on the response to histamine. PMID- 16268986 TI - Right-sided cardiac function in healthy volunteers measured by first-pass radionuclide ventriculography and gated blood-pool SPECT: comparison with cine MRI. AB - BACKGROUND: Right ventricular (RV) function is of interest in an array of cardiopulmonary diseases. First-pass radionuclide ventriculography (FP), gated blood-pool single photon emission tomography (GBPS) and cardiac magnetic resonance imaging (MRI) are three currently used non-invasive methods for evaluation of right-sided cardiac function. The aim of our study was to compare the agreement between these methods when measuring right-sided cardiac function. METHODS: Twenty-four healthy volunteers were included. Mean age was 44 years (range: 25-60) and 29% were females. All participants had FP, GBPS and breath hold cine MRI performed according to standard protocols. RESULTS: Normal ranges for RV ejection fraction (RVEF) defined as mean +/- 2SD were 0.49-0.72, 0.44-0.66 and 0.40-0.69 when measured by MRI, FP and GBPS respectively. Bland-Altman analysis showed a mean difference (bias) between MRI and FP of 0.05 (95% CI: 0.03 0.08) and of 0.06 (95% CI: 0.02-0.10) between MRI and GBPS. No systematic bias was found between FP and GBPS. Normal values for RV end-diastolic volume index (RVEDVI) were 37-95 and 29-91 ml m(-2) when measured by MRI and GBPS respectively. The mean difference between RVEDVI was 6 ml m(-2) (95% CI: 0-11). CONCLUSIONS: (i) Normal values of RVEF differ between MRI, FP and GBPS with wide limits of agreement, accordingly it is difficult to evaluate changes over time if measured by different methods, (ii) RV volumes are in the same range when measured by MRI or GBPS but with wide limits of agreement, and (iii) if MRI is considered gold standard then FP is more accurate than GBPS for RVEF measurements. PMID- 16268987 TI - Recording of dynamic arterial compliance changes during hand elevation. AB - Finger arterial compliance has been studied on the beat-to-beat basis by using the digital arterial pressure and volume waveforms and performing measurements at zero transmural pressure during arm elevation in 11 volunteers. Continuous non invasive finger blood pressure was measured by applying the Finapres monitor and the finger volume pulses were recorded by the UT9201 physiograph by using the photoplethysmographic principle of registration. Estimation of the beat-to-beat finger arterial compliance is based on (i) the recorded volume and pressure wave amplitudes (Vpulse and Ppulse) and (ii) on the calculation of the slope of the pressure-volume relationship from the first derivatives dV/dt and dP/dt of the recorded volume and pressure pulses near the point of the maximum slope. The results of the study demonstrate that the applied two methods similarly (correlation coefficient r = 0.97) describe the changes of the beat-to-beat compliance during hand elevation test. At the same time the second estimate was 18% higher than the first one (P = 0.003). PMID- 16268988 TI - Intestinal circulation, oxygenation and metabolism is not affected by oleic acid lung injury. AB - This study was performed to establish a platform for further studies on effects of ventilatory treatment modalities on the intestines during mechanical ventilation of acute lung injury (ALI). We tested the hypotheses that oleic acid (OA) infusion causes changes in intestinal circulation, oxygenation and metabolism, and that OA is distributed to tissues outside the lung. This was performed as an experimental, prospective and controlled study in an university animal research laboratory. Thirteen juvenile anaesthetized pigs were used in the main study, where seven were given an intravenous infusion of 0.1 ml kg(-1) OA and six served as control (surgery only). In a separate study, four animals were given an intravenous infusion of 0.1 ml kg(-1) (3)H-labelled OA. We measured systemic and mesenteric (portal venous blood flow, jejunal mucosal perfusion) haemodynamic parameters, mesenteric oxygenation (jejunal tissue oxygen tension) and systemic cytokines (tumour necrosis factor-alpha and interleukin-6). We calculated mesenteric lactate flux and mesenteric oxygen delivery, uptake and extraction ratio. In the animals given 3H-OA, we measured 3H-OA in different tissues (lungs, heart, liver, kidney, stomach, jejunum, colon and arterial blood). We found that OA given intravenously is distributed in small amounts to the intestines. This intestinal exposure to OA does not cause intestinal injury when evaluating mesenteric blood flow, metabolism or oxygenation. OA infusion induced a moderate increase in mean pulmonary arterial pressure and a decrease in PaO2/Fraction inspired O2 (P/F) ratio, giving evidence of severe lung injury. Consequently, the OA lung injury model is suitable for studies on intestinal effects of ventilatory treatment modalities during mechanical ventilation of ALI. PMID- 16268990 TI - Female receptivity phenotype of icebox mutants caused by a mutation in the L1 type cell adhesion molecule neuroglian. AB - Relatively little is known about the genes and brain structures that enable virgin female Drosophila to make the decision to mate or not. Classical genetic approaches have identified several mutant females that have a reluctance-to-mate phenotype, but most of these have additional behavioral defects. However, the icebox (ibx) mutation was previously reported to lower the sexual receptivity of females, without apparently affecting any other aspect of female behavior. We have shown that the ibx mutation maps to the 7F region of the Drosophila X chromosome to form a complex complementation group with both lethal and viable alleles of neuroglian (nrg). The L1-type cell adhesion molecule encoded by nrg consists of six immunoglobulin-like domains, five fibronectin-like domains, one transmembrane domain and one alternatively spliced intracellular domain. The ibx strain has a missense mutation causing a glycine-to-arginine change at amino acid 92 in the first immunoglobulin domain of nrg. Defects in the central brain of ibx mutants are similar to those observed in another nrg mutant, central brain deranged(1) (ceb(1)). However, both ceb(1) homozygous and ceb(1)/ibx heterozygous females are receptive. The expression of a transgene containing the non-neural isoform of nrg rescues both the receptivity and the brain structure phenotypes of ibx females. PMID- 16268991 TI - Genetic and environmental influences on Anxious/Depression during childhood: a study from the Netherlands Twin Register. AB - For a large sample of twin pairs from the Netherlands Twins Register who were recruited at birth and followed through childhood, we obtained parental ratings of Anxious/Depression (A/D). Maternal ratings were obtained at ages 3 years (for 9025 twin pairs), 5 years (9222 pairs), 7 years (7331 pairs), 10 years (4430 pairs) and 12 years (2363 pairs). For 60-90% of the pairs, father ratings were also available. Multivariate genetic models were used to test for rater independent and rater-specific assessments of A/D and to determine the genetic and environmental influences on individual differences in A/D at different ages. At all ages, monozygotic twins resembled each other more closely for A/D than dizygotic twins, implying genetic influences on variation in A/D. Opposite sex twin pairs resembled each other to same extent as same-sex dizygotic twins, suggesting that the same genes are expressed in boys and girls. Heritability estimates for rater-independent A/D were high in 3-year olds (76%) and decreased in size as children grew up [60% at age 5, 67% at age 7, 53% at age 10 (60% in boys) and 48% at age 12 years]. The decrease in genetic influences was accompanied by an increase in the influence of the shared family environment [absent at ages 3 and 7, 16% at age 5, 20% at age 10 (5% in boys) and 18% at age 12 years]. The agreement between parental A/D ratings was between 0.5 and 0.7, with somewhat higher correlations for the youngest group. Disagreement in ratings between the parents was not merely the result of unreliability or rater bias. Both the parents provided unique information from their own perspective on the behavior of their children. Significant influences of genetic and shared environmental factors were found for the unique parental views. At all ages, the contribution of shared environmental factors to variation in rater-specific views was higher for father ratings. Also, at all ages except age 12, the heritability estimates for the rater-specific phenotype were higher for mother ratings (59% at age 3 and decreasing to 27% at age 12 years) than for father ratings (between 14 and 29%). Differences between children, even as young as 3 years, in A/D are to a large extent due to genetic differences. As children grow up, the variation in A/D is due in equal parts to genetic and environmental influences. Anxious/Depression, unlike many other common childhood psychopathologies, is influenced by the shared family environment. These findings may provide support for why certain family therapeutic approaches are effective in the A/D spectrum of illnesses. PMID- 16268992 TI - Association between Tph2 gene polymorphism, brain tryptophan hydroxylase activity and aggressiveness in mouse strains. AB - The brain neurotransmitter serotonin is involved in the regulation of aggressive behavior. The main factor determining the brain serotonin level is the activity of the rate-limiting enzyme in the biosynthesis of the neurotransmitter- tryptophan hydroxylase isoform (TPH) 2 encoded by the Tph2 gene. Recently the C1473G single-nucleotide polymorphism in the Tph2 gene was reported. Here we study the C1473G polymorphism in 10 inbred mouse strains (C57BL/6J, AKR/J, DD/He, C3H/HeJ, YT/Y, BALB/cJLac, CC57BR/Mv and A/He) and demonstrate the association of the polymorphism with brain TPH activity and intermale aggressiveness. TPH activity in the midbrain of mice homozygous for the 1473C allele was higher than that in mice carrying 1473G alleles. A close association of the 1473C allele with increased number of attacks towards another male was found. The results support a link between the C1473G polymorphism in Tph2 gene, tryptophan hydroxylase activity and intensity of intermale aggression. PMID- 16268994 TI - Serum antibodies to periodontal pathogens and markers of systemic inflammation. AB - AIM: We examined the relationship between serum antibodies against Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans, and plasma fibrinogen and serum C-reactive protein (CRP) in a nationally representative sample. METHODS: Data on 2,973 participants aged 40 years and older from the third National Health and Nutrition Examination Survey, second phase (1991-1994) were used. Three logistic regression models adjusted for gender, race, educational attainment, diabetes, cigarette smoking, body mass index (BMI), and other inflammatory conditions were constructed, based on three different assumptions: (A) no access to dental/periodontal data; (B) knowledge of number of teeth present but not of clinical periodontal status; and (C) knowledge of both dental and clinical periodontal status. RESULTS: High fibrinogen (>400 mg/dl) was unrelated to P. gingivalis and A. actinomycetemcomitans antibodies in all models. High CRP (>0.4 mg/dl) was related to high antibody levels to P. gingivalis in models A [odds ratios (OR) 1.63, 95% confidence intervals (CI) 1.15-2.32], B (OR 1.69, 95% CI 1.18-2.41), and C (OR 1.58, 95% CI 1.12-2.23). In model C, high CRP was related to >30% extent of attachment loss of >or=3 mm (OR 1.58, 95% CI 1.19-2.08). Antibodies to A. actinomycetemcomitans were not associated with high CRP levels in any model. CONCLUSIONS: High serum titre to P. gingivalis and the presence of periodontal disease are independently related to high CRP levels. PMID- 16268996 TI - Effect of smoking on the gingival capillary density: assessment of gingival capillary density with orthogonal polarization spectral imaging. AB - OBJECTIVES: Microvascular changes because of smoking are frequently presumed in models because of the negative effect of smoking portrayed on the microcirculation. We hypothesized that cigarette smoke might lead to a decrease in gingival capillary density. MATERIALS AND METHODS: Capillary density was assessed with orthogonal polarization spectral (OPS) imaging, a technique using special optics by which a virtual light source is created at a depth of 1 mm within the mucosa. The light is absorbed by haemoglobin, resulting in an image of the capillaries in negative contrast. The gingival capillary density was measured in 20 healthy male dental students with a mean age of 25. Ten of the students were smokers and 10 were non-smokers. In each subject six images of the right maxillary pre-molar region were obtained, and the mean gingival capillary density was determined through the use of K&K software technology. RESULTS: The mean capillary density in smokers was 69.3 +/- 8.9 capillaries per visual field compared with a mean capillary density in non-smokers of 60.6 +/- 5.4 (p=0.33). CONCLUSION: No significant differences were found between the gingival capillary density of smokers and non-smokers. PMID- 16268995 TI - Nicotine inhibits human gingival fibroblast migration via modulation of Rac signalling pathways. AB - AIM: Cigarette smoking is a risk factor in the development of periodontal diseases. In addition, a delayed healing process has been shown in smokers compared with non-smokers after periodontal treatment. Cell migration is a key process of wound healing and it is highly regulated by a variety of signalling pathways. The small G protein, Rac, is necessary for cell migration. Our aim was to determine if nicotine disrupted Rac and its downstream signalling proteins, p21-activated kinase 1/2 (PAK1/2), and p44/42 mitogen-activated protein kinase (MAPK) (extracellular regulated kinase 1/2). MATERIAL AND METHODS: Primary human fibroblasts from healthy gingival tissues were cultured and grown to confluence. Cells were serum starved for 24 h, and then treated with nicotine (0 or 0.5 microM) prior to in vitro wounding. Cell migration was analysed in live cell assays following in vitro wounds. Rac activity, phosphorylation levels of PAK1/2, and p44/42 MAPK were assessed in cultures treated with or without nicotine after multiple wounds. RESULTS: Nicotine decreased cell migration rates by 50% compared with controls. In addition, nicotine altered the activation patterns of Rac and PAK 1/2 and up-regulated p44/42 MAPK. CONCLUSION: Decreased cell migration in periodontal wounds exposed to nicotine may be mediated through the Rac and PAK1/2 signalling pathways. PMID- 16268997 TI - Clinical and microbiological characterization of periodontal abscesses. AB - BACKGROUND/AIM: The knowledge of clinical features, microbial composition and susceptibility to antimicrobials of periodontal abscesses has recently improved. This descriptive clinical and microbiological study provides more information on the characteristics of periodontal abscesses. MATERIALS AND METHODS: Clinical parameters and subgingival samples were examined from 54 subjects presenting 60 periodontal abscesses. Samples were cultured for anaerobic and facultative bacteria, and data were expressed as frequency detection and mean proportion of isolation for microorganisms. Selected isolates of Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans and Prevotella intermedia/nigrescens were used to test susceptibility to amoxicillin, azithromycin, tetracycline and metronidazole. Statistical descriptive analysis was conducted. RESULTS: Most periodontal abscesses were present in patients with ongoing Chronic Periodontitis. Bleeding on probing, tumefaction and suppuration were present in almost all abscesses. Affected teeth were lower anterior teeth, upper anterior teeth and lower molars. The subgingival microbiota was composed of periodontal pathogens such as Fusobacterium spp. (75%), P. intermedia/nigrescens (60%), P. gingivalis (51%) and A. actinomycetemcomitans (30%). Some periodontopathogens showed antimicrobial resistance to tetracycline, metronidazole and amoxicillin, but not to azithromycin. CONCLUSIONS: Periodontal abscesses showed typical clinical features associated with untreated periodontitis, and the organisms identified were important periodontopathic bacteria. Rationale use of antibiotic adjunctive therapy in abscess treatment should be taken into account. PMID- 16268998 TI - Controlled clinical and psychometric studies on the relation between periodontitis and depressive mood. AB - BACKGROUND: Depressive mood is considered a risk factor for the development of periodontitis. OBJECTIVES: Investigation of the relationship between periodontitis and psychopathology utilizing psychometry (both observer- and self rating scales). METHODS: Forty periodontitis patients were compared with 41 age- and sex-matched controls. The percentage of smokers was similar in both groups (30% versus 24.4%). Dental variables included probing depth, clinical attachment loss (CAL), radiographic loss of attachment, papillary bleeding index (PBI) and approximal plaque index (API). Psychometry comprised the Hamilton Depression Scale, the Zung Self-Rating Depression and Anxiety Scales, the von Zerssen Well being and Complaint Scales, the Epworth Sleepiness Scale, the Pittsburgh Sleep Quality Index, the Quality-of-Life Index, crystallized intelligence and the Freiburg Personality Inventory (FPI). RESULTS: Multifactorial analysis of variance demonstrated increased depression and anxiety scores, reduced well being, increased somatic complaints, deteriorated quality of life and introversion in periodontitis. Partial correlation analyses between psychometric measures and dental variables revealed positive correlations of periodontal disease severity/CAL with the depression/anxiety, subjective well-being and complaints scores, and a negative correlation with quality of life. The API was negatively correlated with social orientation, and the CAL was positively correlated with somatic complaints and introversion in the FPI. CONCLUSION: Our clinical-psychometric studies confirm depressive mood as a relevant pathogenetic factor for periodontitis. PMID- 16268999 TI - The influence of stress and anxiety on the response of non-surgical periodontal treatment. AB - AIM: The aim of this study was to evaluate the influence of stress and anxiety on the response to non-surgical periodontal treatment (NPT) in patients with chronic periodontitis. METHOD: Sixty-six patients (mean age 46.1 +/- 8 years) were assigned to three groups: control group, probing pocket depth (PPD) or=4 and 6 mm, n=20. Stress, state anxiety (SA) and trait anxiety (TA) and plaque index (PI), gingival index, PPD and clinical attachment level (CAL) were recorded at baseline and 3 months after NPT. RESULTS: TA scores were different among groups at baseline and after NPT. TA was related to periodontitis at baseline and after NPT. PI was associated with the SA at baseline. The reduction of frequency of CAL >6 mm was correlated with TA after adjusting for confounders. Stressed subjects did not show reduction of frequency of PPD >6 mm (T1), CAL 4-6 mm and CAL >6 mm (T2). CONCLUSIONS: The data suggest an influence of trait of anxiety and stress on the response to NPT. PMID- 16269000 TI - Sonic-powered toothbrushes and reversal of experimental gingivitis. AB - OBJECTIVE: To compare two sonic toothbrushes in relation to the reversal of experimental gingivitis. MATERIALS AND METHODS: Subjects refrained from brushing mandibular teeth for 21 days. During a 4-week treatment phase, the right or left side of the mouth was brushed with either the Sonic Complete (SC) or Sonicare Elite (SE) toothbrush as randomly allocated. Plaque and gingivitis were assessed on day 0, after 21 days of no oral hygiene and after 1, 2 and 4 weeks of brushing twice daily. RESULTS: Thirty-four subjects provided evaluable data. The experimentally induced gingivitis (EIG) resulted in higher bleeding and plaque scores compared with day 0. The mean plaque scores at day 21 changed from 3.09 to 1.30 for the SC, and from 3.02 to 1.21 for the SE. At the end of the treatment period, there was no significant difference between the two brushes. The mean bleeding scores changed from 1.87 (day 21) to 0.97 for the SC, and from 1.83 to 0.92 for the SE. For the assessments at 1, 2 and 4 weeks post-EIG, both brushes showed a significant decrease in bleeding scores. There were no statistically significant differences between brushes. PMID- 16269001 TI - Wear of dentine in vitro by toothpaste abrasives and detergents alone and combined. AB - AIM: To measure in vitro the abrasion of dentine by toothpaste detergents and abrasives alone and combined. MATERIALS AND METHODS: Detergents used were tego betain, sodium lauryl sulphate (SLS), adinol and pluronic diluted to 1% w/v. Abrasives were three artificial silicas, tixosil 73 and 123 and Zeodent 113, and calcium carbonate used at 2.5% w/v. Flat human dentine specimens were brushed with aqueous detergent solutions or abrasive slurries, detergent abrasive slurries and water for 20,000 brush strokes. Dentine loss was measured by non contacting profilometry at 10,000 and 20,000 strokes. Silica particle size distribution was measured by laser deflection. RESULTS: Loss of dentine occurred with all detergents, abrasives and detergent abrasion combinations, but was not linear with number of brush strokes. Water appeared to remove the smear layer only, but all detergents exceeded the predicted smear layer thickness. The silica abrasives differed in abrasion properties despite similar particle size distribution. Different detergents modulated the abrasives actions in mainly positive or mainly negative directions. CONCLUSIONS: Detergents appear able to attack the dentine surface to produce wear. Abrasives vary considerably in wear produced under similar conditions. Detergents modulate the effect of abrasives in a way that may reflect the rheological properties of the mixture. PMID- 16269002 TI - Methyl cellulose gel obstructed bone formation by GBR: an experimental study in rats. AB - AIM: To evaluate whether bone formation under Teflon capsules may be enhanced by concomitant implantation of recombinant human platelet-derived growth factor BB/insulin-like growth factor-I (rhPDGF-BB/IGF-I) incorporated into a methyl cellulose gel. MATERIALS AND METHODS: Fifty-five male 6-month-old albino rats of the Wistar strain were used in the study. The lateral aspect of the mandibular ramus was exposed on both sides of the jaw. In 70 sites, the periosteum was removed from the ramus, leaving the bone denuded, while in 35 sites, it was preserved. On 10 non-periosteal (P-) sites and five periosteal (P+) sites, an empty rigid teflon capsule (d=7 mm), serving as control, was placed on the ramus. In the 40 test animals, the capsule placed on the one side of the jaw was filled at random with one of three different concentrations (1,200, 600, 150 microg/ml) of rhPDGF-BB/IGF-I gel. The capsules placed on the contralateral side of the jaw contained a placebo methyl cellulose gel. Each growth factor group, defined according to the gel concentration, and the placebo group contained 10 capsules placed on the P- side and five capsules placed on the P+ side. Two months after surgery, all animals were sacrificed. RESULTS: Histologic analysis revealed that in the non-filled control capsules, the amount of new bone including the bone marrow was 29.9% and 39.7% of the capsule area on the P- and P+ sides, respectively. In the test capsules with the growth factor gel and placed on the P sides, the amounts of new bone ranged from 5.6% to 6.3%, which were similar (p>0.05) to that formed in the capsules filled with the methyl cellulose gel (5.5%). New bone formation was larger in the capsules on the P+ sides than in those on the P- sides but was similar in the capsules with different growth factor concentrations (range 17.9-19.6%) and in those with placebo gel (21.0%). In all groups, the carrier gel was poorly absorbed and occupied most of the capsules. CONCLUSION: Local application of a methyl cellulose gel obstructed bone formation under Teflon capsules placed adjacent to uninjured cortical bone in the mandibular ramus of rats. These data suggest that another material should be utilized to deliver growth factors under Teflon membranes for guided bone regeneration. PMID- 16269003 TI - Effect of Bio-Oss with or without platelet-derived growth factor on bone formation by "guided tissue regeneration": a pilot study in rats. AB - AIM: To investigate the effect of Bio-Oss with and without the local application of recombinant human platelet-derived growth factor (rhPDGF-BB) on bone formation under Teflon capsules. MATERIALS AND METHODS: Eight male, 6-month-old, Wistar strain rats were used in the study. In each animal, the lateral aspect of the mandibular ramus was exposed and small perforations were produced in the bone. A rigid, non-porous hemispherical teflon capsule (diameter 7 mm) was placed on the ramus in both sides of the animals. The capsule placed on the one side of the jaw was filled with Bio-Oss granules soaked in a solution of PDGF-BB (20 microg/capsule) and autogenous blood prior to placement. The capsules placed on the other side of the jaw were filled with Bio-Oss granules soaked in autogenous blood only (controls). Four rats were sacrificed after 3 months and the remaining four after 5 months. Undecalcified sections containing the capsule and surrounding tissues were prepared and analysed in the microscope. RESULTS: Histologic analysis revealed limited amounts of bone formation. Most of the space underneath the capsules was occupied by Bio-Oss particles surrounded by fibrovascular connective tissue. Given the small sample size statistical analysis was not possible, however, the mean amount of mineralized new bone in the control group (20.8%) appeared to be larger than that in the test group (6.7%). After 5 months the amount of newly formed bone appeared similar in the two groups (23.0% test, 26.0% controls). The Bio-Oss particles occupied between 31.4% and 41.1% of the capsule area at 3 months and between 34.0% and 34.7% at 5 months. Only particles adjacent to the mandibular ramus were incorporated in newly formed bone. CONCLUSION: Limited bone formation was present in the capsules grafted with Bio-Oss with or without the growth factor. PMID- 16269004 TI - Post-translational excision of the carboxyl-terminal segment of CaM kinase phosphatase N and its cytosolic occurrence in the brain. AB - Ca2+/Calmodulin-dependent protein kinase (CaM kinase) phosphatase, occurring in the cytoplasm of all tissues, dephosphorylates and thereby deactivates multifunctional CaM kinases, such as CaM kinases I, II and IV. In contrast, CaM kinase phosphatase N has been reported to occur almost exclusively in the brain and to be localized in the nucleus in the transfected COS-7 cells, as examined immunocytochemically with antibodies against the carboxyl-terminal segment of the enzyme, indicating its involvement in the deactivation of CaM kinase IV. Here, we show that the majority of the naturally occurring CaM kinase phosphatase N in the brain exists not in the intact form of the enzyme (83.4 kDa) but in a form (61.1 kDa) in which the carboxyl-terminal segment containing nuclear localization signals is deleted, and that it is present mostly in the cytoplasm but a little in the nucleus throughout the central nervous system, although occurring mostly in the nucleus in some large neurons. Strong immunostaining of the enzyme was also observed at postsynaptic density. These findings suggest that CaM kinase phosphatase N is involved in the regulation of not only CaM kinase IV but also CaM kinases II and I. PMID- 16269005 TI - The beta-amyloid peptide of Alzheimer's disease decreases adhesion of vascular smooth muscle cells to the basement membrane. AB - Cerebral amyloid angiopathy (CAA) is a major feature of Alzheimer's disease pathology. In CAA, degeneration of vascular smooth muscle cells (VSMCs) occurs close to regions of the basement membrane where the amyloid protein (Abeta) builds up. In this study, the possibility that Abeta disrupts adhesive interactions between VSMCs and the basement membrane was examined. VSMCs were cultured on a commercial basement membrane substrate (Matrigel). The presence of Abeta in the Matrigel decreased cell-substrate adhesion and cell viability. Full length oligomeric Abeta was required for the effect, as N- and C-terminally truncated peptide analogues did not inhibit adhesion. Abeta that was fluorescently labelled at the N-terminus (fluo-Abeta) bound to Matrigel as well as to the basement membrane heparan sulfate proteoglycan (HSPG) perlecan and laminin. Adhesion of VSMCs to perlecan or laminin was decreased by Abeta. As perlecan influences VSMC viability through the extracellular signal-regulated kinase (ERK)1/2 signalling pathway, the effect of Abeta1-40 on ERK1/2 phosphorylation was examined. The level of phospho-ERK1/2 was decreased in cells following Abeta treatment. An inhibitor of ERK1/2 phosphorylation enhanced the effect of Abeta on cell adhesion. The studies suggest that Abeta can decrease VSMC viability by disrupting VSMC-extracellular matrix (ECM) adhesion. PMID- 16269006 TI - Action of extracellular divalent cations on native alpha-amino-3-hydroxy-5 methylisoxazole-4-propionate (AMPA) receptors. AB - The effects of divalent cations on Ca2+-impermeable containing (GluR2 subunit) MPA receptors of hippocampal pyramidal neurones isolated from rat brain was studied using patch-clamping. Ca2+, Mg2+, Mn2+, Co2+, Ni2+ and Zn2+ inhibited currents induced by kainate and glutamate. Inhibition was fast, reversible and voltage independent. The rank order of activities was Ni2+ > Zn2+ > Co2+ > Ca2+ > Mn2+ > Mg2+. Cyclothiazide (0.1 mm) significantly reduced inhibition by divalent cations and 6, 7 dinitroquinoxaline-2.3-dione (DNQX). However, high concentrations of Ni2+ and DNQX inhibited AMPA receptors even in the presence of cyclothiazide. The inhibitory effect of divalent cations as well as DNQX was counteracted by an increase in agonist concentration. In the presence of divalent cations the EC50 values of kainate and glutamate were increased, but the maximal response was not changed. An increase in agonist concentration induced a parallel shift in the concentration-inhibition curve for a divalent cation. These data suggest a competitive-like type of inhibition. However, an increase in agonist concentration reduced the inhibitory action of Ni2+ less than that of DNQX. This gave evidence against direct competition between divalent cations and AMPA receptor agonists. A 'complex-competition' hypothesis was proposed to explain the inhibitory action of divalent cations; it is suggested that divalent cations form ion-agonist complexes, which compete with free agonist for agonist-binding sites on AMPA receptors. PMID- 16269007 TI - Striatal neuroadaptation and rescue of locomotor deficit by L-dopa in aphakia mice, a model of Parkinson's disease. AB - Preferential neurodegeneration of dopaminergic neurons in the ventral substantia nigra of the midbrain is a hallmark of Parkinson's disease. The homeobox transcription factor Pitx3 is similarly and selectively expressed in the same neurons. Pitx3 deficiency in a natural mouse mutant, the aphakia mouse, was correlated with the loss of these neurons and with a deficit in locomotor activity. We now report that the locomotor deficit of aphakia mice is established by 40 days of age and that it can be rescued by injection of l-dopa. We further show that downstream striatal correlates of the midbrain neuronal losses in aphakia mice, as assessed by dopamine transporter binding and expression of dopamine receptors, enkephalin, dynorphin and neurotensin, are highly similar to neuroadaptive responses observed following rapid neurodegeneration induced by neurotoxin administration in adult animals or following the progressive neurodegenerative processes as seen in Parkinson patients. Taken collectively, these data support the idea that the aphakia mice represent a selective model of dopaminergic deficiency that closely resembles the midbrain and striatal neuropathology associated with Parkinson's disease, and this suggests that these mice are a good model to assess therapies for Parkinson's disease as well as to understand the susceptibility of these neurons to neurodegeneration. PMID- 16269008 TI - Fibronectin promotes brain capillary endothelial cell survival and proliferation through alpha5beta1 and alphavbeta3 integrins via MAP kinase signalling. AB - We showed previously that blood vessel maturation in the CNS is associated with a developmental switch in brain capillary endothelial cells (BCEC), from fibronectin signalling during angiogenesis to laminin signalling in the adult. To investigate the functional significance of this switch, we have examined the response of BCEC to different extracellular matrix (ECM) proteins. This showed that BCEC proliferation was significantly promoted by fibronectin (28.2 +/- 4.0%) and by vitronectin (14.8 +/- 2.1%) compared with uncoated glass (7.2 +/- 0.7%), while BCEC survival was significantly promoted by fibronectin (1130 +/- 131 cells), vitronectin (830 +/- 63 cells), collagen IV (703 +/- 77 cells) and laminin (680 +/- 34 cells) compared with the uncoated glass (367 +/- 48 cells). Biochemical studies showed that BCEC express a limited repertoire of integrins, including the beta1 integrins, alpha3beta1, alpha5beta1 and alpha6beta1, and the alphavbeta3 integrin. Function-blocking studies showed that the response to fibronectin was mediated equally by the alpha5beta1 and alphavbeta3 integrins. Analysis of signalling pathways revealed that fibronectin stimulated activation of the p44/p42 MAP kinase signalling pathway and pharmacological inhibitors of this pathway blocked BCEC proliferation on fibronectin. Taken together, these findings show that fibronectin exerts a strong angiogenic influence on endothelial cells (EC) in the CNS, and that this is mediated through the alpha5beta1 and alphavbeta3 integrins via MAP kinase signalling. In addition to a fundamental role in development, these findings may also have implications in pathological conditions of the CNS where fibronectin is re-expressed. PMID- 16269010 TI - Molecular and pharmacological characterization of muscarinic receptors in retinal pigment epithelium: role in light-adaptive pigment movements. AB - Muscarinic receptors are the predominant cholinergic receptors in the central and peripheral nervous systems. Recently, activation of muscarinic receptors was found to elicit pigment granule dispersion in retinal pigment epithelium isolated from bluegill fish. Pigment granule movement in retinal pigment epithelium is a light-adaptive mechanism in fish. In the present study, we used pharmacological and molecular approaches to identify the muscarinic receptor subtype and the intracellular signaling pathway involved in the pigment granule dispersion in retinal pigment epithelium. Of the muscarinic receptor subtype-specific antagonists used, only antagonists specific for M1 and M3 muscarinic receptors were found to block carbamyl choline (carbachol)-induced pigment granule dispersion. A phospholipase C inhibitor also blocked carbachol-induced pigment granule dispersion, and a similar result was obtained when retinal pigment epithelium was incubated with an inositol trisphosphate receptor inhibitor. We isolated M2 and M5 receptor genes from bluegill and studied their expression. Only M5 was found to be expressed in retinal pigment epithelium. Taken together, pharmacological and molecular evidence suggest that activation of an odd subtype of muscarinic receptor, possibly M5, on fish retinal pigment epithelium induces pigment granule dispersion. PMID- 16269011 TI - Fibroblast growth factor 2 applied to the optic nerve after axotomy up-regulates BDNF and TrkB in ganglion cells by activating the ERK and PKA signaling pathways. AB - Application of basic fibroblast growth factor (FGF-2) to the optic nerve after axotomy promotes the survival of retinal ganglion cells (RGCs) in the frog, Rana pipiens. Here we investigate the effects of FGF-2 treatment upon the synthesis of brain-derived neurotrophic factor (BDNF) and its receptor, tyrosine receptor kinase B (TrkB). Axotomy alone increased the amounts of BDNF and TrkB mRNA in RGCs after 1 week and 48 h, respectively; FGF-2 treatment to the nerve accelerated and increased this up-regulation of both. FGF-2 also increased the amounts of phosphorylated cAMP response element binding protein (pCREB) in the retina. Blocking extracellular-regulated kinase (ERK) activation with PD98059 or U0126 prevented the FGF-2-induced up-regulation of BDNF transcription but had no effect on TrkB. However, blocking protein kinase A (PKA) with H89 or Rp-8-Cl cAMPS reduced the up-regulation of both BDNF and TrkB, and reduced pCREB. In addition, H89 inhibited ERK activation, indicating cross-talk between the pathways. Finally, axonal application of blocking antibody against the FGF receptor 1 (FGFR1) prevented the FGF-2-induced up-regulation of BDNF and TrkB. Our results suggest that FGF-2 acts on RGCs via FGFR1, activating the ERK pathway and CREB to increase BDNF synthesis, and PKA and CREB to increase TrkB synthesis. PMID- 16269012 TI - Immunoisolation of two synaptic vesicle pools from synaptosomes: a proteomics analysis. AB - The nerve terminal proteome governs neurotransmitter release as well as the structural and functional dynamics of the presynaptic compartment. In order to further define specific presynaptic subproteomes we used subcellular fractionation and a monoclonal antibody against the synaptic vesicle protein SV2 for immunoaffinity purification of two major synaptosome-derived synaptic vesicle containing fractions: one sedimenting at lower and one sedimenting at higher sucrose density. The less dense fraction contains free synaptic vesicles, the denser fraction synaptic vesicles as well as components of the presynaptic membrane compartment. These immunoisolated fractions were analyzed using the cationic benzyldimethyl-n-hexadecylammonium chloride (BAC) polyacrylamide gel system in the first and sodium dodecyl sulfate-polyacrylamide gel electrophoresis in the second dimension. Protein spots were subjected to analysis by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI TOF MS). We identified 72 proteins in the free vesicle fraction and 81 proteins in the plasma membrane-containing denser fraction. Synaptic vesicles contain a considerably larger number of protein constituents than previously anticipated. The plasma membrane-containing fraction contains synaptic vesicle proteins, components of the presynaptic fusion and retrieval machinery and numerous other proteins potentially involved in regulating the functional and structural dynamics of the nerve terminal. PMID- 16269013 TI - NMDA-induced retinal injury is mediated by an endoplasmic reticulum stress related protein, CHOP/GADD153. AB - We investigated the role of an endoplasmic reticulum stress-associated protein, CHOP/GADD153, after NMDA-induced mouse retinal damage. After injection of NMDA into the vitreous, TUNEL-positive cells were detected in the retinal ganglion cell layer (GCL) and inner nuclear layer (INL) at 6 h after NMDA injection, and these gradually increased in number up to 24 h. Analysis by real-time RT-PCR revealed that CHOP mRNA was induced by about 3-fold, at 2 h after NMDA injection. Immunoreactivity for the CHOP protein was intense in cells of the GCL following NMDA treatment. Immunoblot analysis showed that NMDA injection increased the expression of CHOP protein in the retina. Compared with wild-type mice, CHOP/ mice were more resistant to NMDA-induced retinal cell death as determined by TUNEL assay. At 7 days after NMDA treatment, the thickness of the inner plexiform layer and INL were larger in CHOP/ mice than in wild-type mice. The number of residual cells in the GCL following NMDA treatment was significantly higher in CHOP/ mice than in wild-type mice. In conclusion, CHOP is induced in mouse retina by NMDA treatment, and CHOP/ mice are more resistant to NMDA-induced retinal damage, suggesting that CHOP plays an important role in NMDA-induced retinal cell death. PMID- 16269014 TI - Quantitative measurement of serotonin synthesis and sequestration in individual live neuronal cells. AB - Synthesis and subsequent sequestration into vesicles are essential steps that precede neurotransmitter exocytosis, but neither the total neurotransmitter content nor the fraction sequestered into vesicles have been measured in individual live neurons. We use multiphoton microscopy to directly observe intracellular and intravesicular serotonin in the serotonergic neuronal cell line RN46A. We focus on how the relationship between synthesis and sequestration changes as synthesis is up-regulated by differentiation or down-regulated by chemical inhibition. Temperature-induced differentiation causes an increase of about 60% in the total serotonin content of individual cells, which goes up to about 10 fmol. However, the number of vesicles per cell increases by a factor of four and the proportion of serotonin sequestered inside the vesicles increases by a factor of five. When serotonin synthesis is inhibited in differentiated cells and the serotonin content goes down to the level present in undifferentiated cells, the sequestered proportion still remains at this high level. The total neurotransmitter content of a cell is, thus, an unreliable indicator of the sequestered amount. PMID- 16269015 TI - New insights into the pathogenesis of the hyperimmunoglobulinaemia E syndrome. PMID- 16269016 TI - VEGF-A, VEGF-D and VEGF-D(DeltaNDeltaC) induced intimal hyperplasia in carotid arteries. AB - BACKGROUND: The role of vascular endothelial growth factors (VEGFs) in intimal hyperplasia and atherogenesis remains unknown. Several studies have suggested that some members of the VEGF family reduce intimal hyperplasia, but others have proposed that VEGFs accelerate restenosis and atherosclerosis. This investigation conducted a comparative study with adenoviruses encoding different VEGFs in a rabbit carotid artery collar model of intimal hyperplasia in order to analyze the role of VEGFs in the formation of intimal hyperplasia. MATERIALS AND METHODS: Intimal hyperplasia was induced in the carotid arteries of cholesterol fed New Zealand White rabbits using a silastic collar. Adenoviral vectors encoding VEGF A, VEGF-B, VEGF-C, VEGF-C(DeltaNDeltaC), VEGF-D and VEGF-D(DeltaNDeltaC) were delivered to the adventitia using the collar as a gene delivery device. Adeno LacZ was used as a control. RESULTS: A significant (P < 0.01) increase in the intima/media ratio was observed in the arteries transduced with VEGF-A, VEGF-D and VEGF-D(DeltaNDeltaC). There was a significant increase in the number of proliferating cells in the adventitia, media and intima of the VEGF-A, VEGF-D and the VEGF-D(DeltaNDeltaC) transduced arteries. The majority of medial smooth muscle cells in these arteries had a synthetic phenotype. The presence of matrix metalloproteinase-2 (MMP-2) and MMP-9 in the VEGF-A, VEGF-D and the VEGF D(DeltaNDeltaC) transduced arteries was significantly increased. A significant positive correlation was observed between adventitial angiogenesis and intimal hyperplasia. CONCLUSIONS: Adventitial delivery of adenoviruses encoding VEGF-A, VEGF-D and VEGF-D(DeltaNDeltaC) increased intimal hyperplasia in the rabbit collar model. Adventitial angiogenesis correlated positively with the intimal hyperplasia. These results indicated that efficient adventitial production of VEGF-A, VEGF-D and VEGF-D(DeltaNDeltaC) can cause thickening of the inner layer of the artery in rabbits. PMID- 16269017 TI - Combined cord blood stem cells and gene therapy enhances angiogenesis and improves cardiac performance in mouse after acute myocardial infarction. AB - BACKGROUND: Gene and stem cell therapies hold promise for the treatment of ischaemic cardiovascular disease. However, combined stem cell and angiogenic growth factor gene therapy for acute ischaemic myocardium has not been previously reported. This study hypothesized that combined stem cell and gene therapy would not only augment new vessels formation but also improve myocardial function in acute ischaemic myocardium. METHODS: Human angiopoietin-1 (Ang1) cDNA and VEGF(165) cDNA were ligated into AAV vector. The purified CD34(+) cells were obtained from human umbilical cord blood samples. Cord blood CD34(+) cells were transduced with AAV vector encoding either the human Ang1 (AAV-Ang1) or VEGF(165) (AAV-VEGF) cDNA alone, or both (AAV-Ang1 plus VEGF). Immediately after ligation of the left anterior descending coronary artery in male SCID mice, culture expanded CD34(+) cells transduced with AAV-Ang1, AAV-VEGF or AAV-Ang1 plus VEGF were injected intramyocardially at the left anterior free wall. RESULTS: Western blot showed that Ang1 and VEGF protein expressions were enhanced in the CD34(+)cells transduced with AAV-Ang1 and AAV-VEGF, respectively. Infarct size significantly decreased and capillary density significantly increased after treatment with CD34(+)/AAV-Ang1 plus VEGF when compared with treatment by CD34(+) only. Combined therapy with CD34(+) and AAV-Ang1, CD34(+) and AAV-VEGF, CD34(+) and AAV-Ang1 plus VEGF, all showed significantly higher cardiac performance in echocardiography than the therapy with CD34(+) alone 4 weeks after myocardial infarction. CONCLUSIONS: Combined therapy with human umbilical cord blood CD34(+) cells and both Ang1 and VEGF genes reduced infarct size, attenuated the progression of cardiac dysfunction and increased capillary density in acute myocardial infarction in mice. PMID- 16269018 TI - Erythrocyte transport efficacy of human blood: a rheological point of view. AB - BACKGROUND: Large-scale epidemiological studies have demonstrated that both anaemia and polycytaemia are independent cardiovascular risk factors. This was substantiated by the Framingham study, which demonstrated a U-shaped relation between haemoglobin concentration and mortality. It was previously noted that delineating the corresponding haematocrit/blood viscosity ratios in the function of haematocrit provided a distribution of an inverted U-shaped curve. The peak appeared physiologically important because it denotes a healthy balance between a relatively high oxygen binding capacity and a moderately low blood viscosity. It was the aim of this study to examine the mathematical relationship between the haematocrit and haematocrit/blood viscosity ratio. MATERIALS AND METHODS: In a retrospective study, the haemorheological data of 32 healthy controls, 52 outpatients with hyperlipidaemia and 120 outpatients with Raynaud's disease were analyzed. Whole blood viscosity was measured with Hevimet 40 capillary viscometer at 37.0 degrees C and at shear rates of 10 s(-1), 90 s(-1) and 200 s(-1). RESULTS: Haematocrit/blood viscosity ratios in the function of haematocrit values showed a Gaussian association in the healthy subjects, hyperlipidaemic and Raynaud's disease outpatient groups. Peak values (i.e. the rheologically optimal haematocrit) were shear-rate and group dependent and were found at 44.3%, 43.5% and 38.3% in controls, hyperlipidaemic and Raynaud's disease patients, respectively, at a shear rate of 90 s(-1). CONCLUSIONS: This is one of the first reports in which a theoretically optimal haematocrit value was determined using the haematocrit/blood viscosity ratio. Further studies are needed to examine the potential clinical usefulness of this approach. PMID- 16269019 TI - Cognitive and physiological effects of Omega-3 polyunsaturated fatty acid supplementation in healthy subjects. AB - BACKGROUND: It has been reported that Omega-3 fatty acids may play a role in nervous system activity and that they improve cognitive development and reference memory-related learning, increase neuroplasticity of nerve membranes, contribute to synaptogenesis and are involved in synaptic transmission. The aim of this study was to examine the effects of Omega-3 supplementation on some cognitive and physiological parameters in healthy subjects. MATERIALS AND METHODS: Subjects were tested at the beginning of the experiment and after 35 days. In this period they were supplemented with Omega-3 polyunsaturated fatty acids. A group was supplemented with olive oil (placebo). Tests involving different types of attention were used, i.e. Alert, Go/No-Go, Choice and Sustained Attention. For each test, the reaction time, the event-related potentials by electroencephalogram (EEG) and the electromyography (EMG) of the forefinger flexor muscle were recorded. The Profile of Mood States test (POMS) was also administered. RESULTS: Blood analyses showed that after Omega-3 supplementation the arachidonic acid/eicosapentaenoic acid ratio (AA/EPA) was strongly reduced. The mood profile was improved after Omega-3 with increased vigour and reduced anger, anxiety and depression states. This was associated with an effect on reactivity with a reduction of reaction time in the Go/No-Go and Sustained Attention tests. The latency of EMG activation was concomitantly reduced in the same tests plus Choice. An EEG frequency shift towards the theta and alpha band were recorded in all the tests after Omega-3. CONCLUSIONS: Omega-3 supplementation is associated with an improvement of attentional and physiological functions, particularly those involving complex cortical processing. These findings are discussed in terms of the influence of Omega-3 on the central nervous system. PMID- 16269021 TI - Recombinant human erythropoietin: effects on frataxin expression in vitro. AB - BACKGROUND: Friedreich's ataxia (FRDA) is a neurodegenerative disorder caused by decreased expression of the protein frataxin, recently described to be an iron chaperone for the assembly of iron-sulphur clusters in the mitochondria, causing iron accumulation in mitochondria, oxidative stress and cell damage. Searching for compounds that could possibly influence frataxin expression, we found that the cytokine recombinant human erythropoietin (rhuEPO) significantly increases frataxin expression by a still unknown mechanism. MATERIALS AND METHODS: Isolated lymphocytes from FRDA patients, isolated human cardiac cells (fibroblasts and myocytes) from patients undergoing heart transplantation and P19 mouse cells (neuronal typ), were incubated with different concentrations of rhuEPO, and immunoblot was carried out for the detection of frataxin. RESULTS: We show for the first time that the cytokine recombinant human erythropoietin (rhuEPO) can, additionally to its reported neuro- and cardioprotective properties, increase frataxin expression in vitro. We show that rhuEPO significantly increases frataxin expression in primary lymphocytes from patients with Friedreich's ataxia. Further we show that rhuEPO can also increase frataxin expression in many other cell types; among them the most affected cell types in FRDA such as neurones and cardiac cells. CONCLUSIONS: Our results provide a scientific basis for further studies examining the effectiveness of this agent for the treatment of FRDA patients. PMID- 16269020 TI - Four peptide hormones' specific decrease (up to 97%) of human prostate carcinoma cells. AB - BACKGROUND: Mortality from prostate cancer remains a significant problem with current treatment(s), with an expected 30 350 deaths from prostate cancer in 2005. Long-acting natriuretic peptide, vessel dilator, kaliuretic peptide and atrial natriuretic peptide have significant anticancer effects in breast and pancreatic adenocarcinomas. Whether these effects are specific and whether they have anticancer effects in prostate adenocarcinoma cells has not been determined. MATERIALS AND METHODS: These peptide hormones were evaluated to determine if they have specific anticancer effects in human prostate adenocarcinomas. RESULTS: Dose response curves revealed a significant (P < 0.05) decrease in human prostate cancer number with each tenfold increase in the concentration from 1 microM to 1000 microM (i.e. 1 mM) of these four peptide hormones. There was a 97.4%, 87%, 88% and 89% (P < 0.001 for each) decrease in prostate cancer cells secondary to vessel dilator, long-acting natriuretic peptide, kaliuretic peptide and atrial natriuretic peptide, respectively, at their 1-mM concentrations within 24 h, without any proliferation in the 3 days following this decrease. These same hormones decreased DNA synthesis from 68% to 89% (P < 0.001). When utilized with their respective antibodies their ability to decrease prostate adenocarcinoma cells or inhibit their DNA synthesis was completely blocked. Western blots revealed that for the first time natriuretic peptide receptors (NPR) A- and C- were present in prostate cancer cells. CONCLUSIONS: These results indicate that these peptide hormones' anticancer effects are specific. Furthermore, they have very potent effects of eliminating up to 97% of prostate cancer cells within 24 h of treatment. PMID- 16269022 TI - Severely impaired IL-12/IL-18/IFNgamma axis in patients with hyper IgE syndrome. AB - BACKGROUND: Hyperimmunoglobulin E syndrome (HIES) is an inborn disorder characterized by recurrent skin and respiratory tract infections, skeletal abnormalities, chronic eczema and elevated serum IgE. The nature of the host defence defect has yet to be established. The aim of this study was to investigate whether activation of the interleukin-12/interleukin-18/interferon gamma axis, known to be crucial for the activation of cellular immune responses, is impaired in patients with HIES. MATERIALS AND METHODS: Cytokine production capacity of seven HIES patients and seven healthy controls was investigated in whole-blood cultures stimulated with heat-killed Staphylococcus aureus, Candida albicans, or a combination of interleukin (IL)-12/IL-18. RESULTS: Interferon (IFN)gamma production, in addition to IFNgamma/IL-10 ratios, was 10-30-fold lower in the HIES patients compared with the healthy volunteers. In contrast TNF, IL 1beta and IL-8 production was normal. CONCLUSIONS: These data revealed a severe dysbalance towards a Th2 phenotype in HIES patients which is likely to contribute to the specific pattern of infection susceptibility characteristic to HIES. PMID- 16269024 TI - Number V Oral lichen planus: clinical features and management. AB - Oral lichen planus (OLP) is a relatively common chronic inflammatory disorder affecting stratified squamous epithelia. Whereas in the majority of instances, cutaneous lesions of lichen planus (LP) are self-limiting and cause itching, oral lesions in OLP are chronic, rarely undergo spontaneous remission, are potentially premalignant and are often a source of morbidity. Current data suggest that OLP is a T cell-mediated autoimmune disease in which auto-cytotoxic CD8+ T cells trigger apoptosis of oral epithelial cells. The characteristic clinical aspects of OLP may be sufficient to make a correct diagnosis if there are classic skin lesions present. An oral biopsy with histopathologic study is recommended to confirm the clinical diagnosis and mainly to exclude dysplasia and malignancy. The most commonly employed and useful agents for the treatment of lichen planus (LP) are topical corticosteroids but other newer agents are available. PMID- 16269025 TI - Human papillomavirus and disease mechanisms: relevance to oral and cervical cancers. AB - Oral squamous cell carcinoma (OSCC) is the sixth most common malignancy and is a major cause of cancer morbidity and mortality worldwide. Carcinoma of the uterine cervix is the most common female malignancy in the world. While cervical cancer is a worldwide disease, oral cancer has the highest incidence in developing countries, especially among tobacco and alcohol users and betel quid chewers. A strong association of cervical and oral cancer with high-risk human papillomavirus (HPV) 16 and 18 infections underlines the importance of the virus in the pathogenesis of these squamous cell carcinomas. Functionally high-risk HPV infection contributes to carcinogenesis and tumor progression predominantly through the actions of two viral oncogenes, E6 and E7. The E6 and E7 genes have been studied in different patient populations and a number of variants have been described. More than 40 variants have been classified and may be related to differences in progression of squamous intraepithelial lesions. The transcription factor, NFkappaB and its activation pathways are frequently targeted by viruses and aberrant constitutive activation of NFkappaB is frequently found in human tumors of diverse tissue origin. Diet-gene interactions are also likely to contribute considerably to the observed inter-individual variations in HPV associated cancer risk, in response to exposures to the nutritional factors that have the potential to promote or protect against cancer. PMID- 16269026 TI - Fructooligosaccharide consumption improves the decreased dentin formation and mandibular defects following gastrectomy in rats. AB - OBJECTIVE: We examined the effects of fructooligosaccharides (FOS) consumption on gastrectomy-evoked osteopenia and disorders of dentin formation in rats. MATERIALS AND METHODS: Male Sprague-Dawley rats (n = 28, 35-day old) were equally divided into two groups; sham-operated and gastrectomized, and sham-operation or total gastrectomy was performed. Four weeks after each surgery, the rats were divided into two sub-groups (n = 7 each); with or without 7.5% FOS-feeding for 6 weeks. Backscattered electron images of the mandibular sections were taken to calculate trabecular bone area, cortical bone area and total scan area. Thereafter, the dentin formation rate in maxilla were calculated using a fluorescent microscope. RESULTS: Trabecular bone area and cortical bone area in GX rats were markedly decreased. FOS-feeding significantly counteracted this reduction, but not to the level seen in sham-operated rats. Total scan area in gastrectomized groups was significantly decreased. The dentin formation rate was not statistically different among the groups, except the gastrectomized group. CONCLUSION: These results suggest that FOS consumption partially restored osteopenia and almost completely restored the reduction in dentin formation following gastrectomy in rats. PMID- 16269028 TI - Oral lesions as indicators of HIV infection among routine dental patients in Lagos, Nigeria. AB - OBJECTIVES: To document the incidental oral lesions of human immunodeficiency virus (HIV) infection, the pattern and frequency of the lesions based on clinical presentation and oral manifestations in routine dental patients who tested positive in Nigeria. SUBJECTS AND METHODS: The study was conducted at the Oral Diagnosis/Oral Medicine clinic of the Lagos University Teaching Hospital, Lagos, Nigeria between May 2002 and April 2003. During this period, all patients with oral lesions suggestive of HIV/acquired immunodeficiency syndrome (AIDS) as described in the EEC-WHO Classification and diagnostic criteria of oral lesions of HIV were counseled and offered voluntary HIV testing. All the 35 patients who consented and tested positive were included in this study. RESULTS: Of a total of 700 patients 53 patients with oral lesions suggestive of HIV/AIDS were seen, thirty-eight (72%) consented to HIV screening, 15 patients (28%) refused. Thirty five patients (92%), mean age 36 +/- 13 years were confirmed positive for HIV. Oral candidiasis was the commonest lesion seen (43%) the second common being Herpes zoster (23%). Other lesions seen included erythema multiforme in two (6%), facial palsy in two (6%) and oral hairy leukoplakia in one (3%). CONCLUSION: An oral mucosal lesion may be the presenting lesion of HIV/AIDS in routine patients attending the dental clinic. Oral health care workers should practice optimal infection control based on the Centers for Disease Control 'Standard Precautions' guidelines on infection control for all patients to minimize occupational transmission of HIV. PMID- 16269027 TI - Oral avascular bone necrosis associated with chemotherapy and biphosphonate therapy. AB - BACKGROUND: Oral avascular bone necrosis is an important adverse effect of chemotherapy and biphosphate therapy. OBJECTIVE: To report our experience in oral avascular bone necrosis in cancer patients assigned to undergo chemotherapy. PATIENTS AND METHODS: Fourteen patients presenting oral avascular bone necrosis were selected from the clinical files of five Stomatological Clinics in Brazil. Clinical data as well as treatment and prognosis information were obtained from all 14 patients. RESULTS: Twelve patients (86%) were submitted to biphosphonate therapy. The most important symptom was pain, present in all cases, and the mandible was the most common involved site. Most patients (79%) had their conditions managed by antibiotic therapy and surgical debridation; however complete response was achieved in only three cases (21%). CONCLUSION: Avascular bone necrosis is a serious oral side-effect of cancer chemotherapy, particularly in patients using biphosphonates, and antibiotic therapy and surgical debridation were not able to promote complete response in most cases. PMID- 16269030 TI - Intra-oral carcinomas in Maiduguri, north-eastern Nigeria. AB - OBJECTIVE: To document the pattern of intra-oral carcinomas diagnosed in north eastern Nigeria. DESIGN AND SETTING: A retrospective clinical and histopathological review seen in a tertiary care hospital (January 1987-December 2002). MATERIALS AND METHODS: Information on socio-demographic characteristics of patients and details of cancer management were retrieved from the files and histopathology records of cases diagnosed at the hospital. RESULTS: A total of 378 biopsies, 317 primary cancers including 279 carcinomas of the head and neck were diagnosed during the study period. Intra-oral carcinomas constitute 43 (15.4%) of all head and neck carcinomas reported, with no occurrence in children. The overall mean age of occurrence was 51.2 +/- 15.6 years (male = 56.2 +/- 13.7 years; females = 47.5 +/- 16.2 years) and an overall male-female ratio of 3:4. Carcinomas were commonly reported in the palate 19 (44.2%) and lip six (13.9%) and floor of mouth four (9.3%). Squamous cell carcinoma 28 (65.1%), adenoidcystic carcinoma seven (16.3%) and mucoepidermoid carcinoma five (11.6%) were the commonly reported carcinomas. Squamous cell carcinoma was the most common carcinoma in all sites, in the sixth decade of life, of equal gender distribution and commonly reported in users of kola nuts and tobacco. The occupation of patients diagnosed with oral carcinoma and squamous cell carcinoma was farming (50%, 61.5%). All the staged cancers patients (n = 7) reported in the late stages (III/IV) of the disease. The mean interval between symptoms and presentation for the different carcinomas ranged between 9 and 25 months, with the least interval reported for mucoepidermoid carcinoma. CONCLUSIONS: Squamous cell carcinoma was the most common oral carcinoma, commonly reported in the palate, among farmers and in the sixth decade of life. Its occurrence in under 40 year olds is three to six times greater than reported for the USA and Europe and may be associated with poor diet and the habitual use of kola nuts and tobacco. The survival rates of patients diagnosed with intra-oral carcinomas, although not available, would be expectedly low in view of the prognostic indicators recorded in this series. The routine oral cancer screening of each patient, and counselling of patients with high-risk habits, by dentists is recommended to improve the prognosis of the disease. PMID- 16269029 TI - Norwegian LongoVital and recurrent aphthous ulceration: a randomized, double blind, placebo-controlled study. AB - OBJECTIVE: LongoVital (LV) is a herbal-based tablet enriched with the recommended daily doses of vitamins. The present study was undertaken to investigate possible prevention of recurrent aphthous ulceration (RAU) during 4 months daily intake of the Norwegian LV. DESIGN: The study was a placebo-controlled, double-blind, randomized, clinical trial. SUBJECTS: Sixty otherwise healthy patients with at least one attack of minor RAU per 2 months were included in the study. METHODS: After an introduction period (IP) of 60 days, the patients were randomly divided into three groups and given either LV, the herbs of LV only, or placebo. Three test tablets were taken every day together with breakfast for 4 months [tablet period (TP)] and the patients followed up for another 4 months (F-UP). The number of new ulcers (NU) and ulcer-free days (UFD) were observed. RESULTS: Fifty-two patients completed the study. Neither NU nor UFD showed any statistical significant differences between any of the groups in any of the periods. All three groups, however, showed a significant increase in UFD during the first 2 months of TP compared to IP. Within the LV group only, there was a further increase in UFD after 2 months intake of the tablets. The number of NU and UFD decreased significantly in both the LV and the herbal group in F-UP compared with TP. CONCLUSION: Neither the Norwegian LV nor the herbal component alone was superior to placebo in the prevention of RAU. The results, however, indicate that neither the LV nor the herbal group benefited from the treatment. PMID- 16269031 TI - Salivary gland neoplasms in Maiduguri, north-eastern Nigeria. AB - OBJECTIVE: To document the pattern of salivary gland neoplasia in Maiduguri, Nigeria. DESIGN AND SETTING: A retrospective clinical and histopathological review (January 1987-December 2002) of cases diagnosed at a tertiary care hospital. MATERIALS AND METHODS: Information on demographics, diagnosis and cancer management in the hospital were retrieved from biopsy reports and case notes of patients. RESULTS: The palatal (71.9%) and parotid (78.3%) glands were the most common minor and major salivary glands involved, with a benign-malignant ratio of 1:1 and 1.4:1, respectively. Pleomorphic adenoma (44.3%) was the most common salivary gland neoplasm recorded. It was commonly reported in the third decade (mean 30.4 years) and among males (M:F, 1.4:1). Ectopic lesions (17.1%) were reported in the neck, nose and cervical nodes. Mucoepidermoid carcinoma (10.1%) was the most common salivary gland malignancy, occurring in the second and sixth decades; of equal gender distribution and predominantly in the palate (50%). The squamous cell carcinoma (10.9%) and adenoidcystic carcinoma (21.9%) were the most common malignancies in the major and minor glands respectively. There was a higher prevalence of malignancies of the parotid than previously reported for northern Nigeria (P = 0.036). CONCLUSION: Pleomorphic adenoma and mucoepidermoid carcinoma were the most commonly reported benign and malignant neoplasia in this series. The prevalence of mucoepidermoid carcinoma contrasts with reported findings in other African studies. PMID- 16269032 TI - Phenotypic methods and commercial systems for the discrimination between C. albicans and C. dubliniensis. AB - Candida dubliniensis is a recently described Candida species associated with oral candidosis that exhibits a high degree of phenotypic similarity to Candida albicans. However, these species show differences in levels of resistance to antimycotic agents and ability to cause infections. Therefore, accurate clinical identification of C. dubliniensis and C. albicans species is important in order to treat oral candidal infections. Phenotypic identification methods are easy-to use procedures for routine discrimination of oral isolates in the clinical microbiology laboratory. However, C. dubliniensis may be so far underreported in clinical samples because most currently used identification methods fail to recognize this yeast. Phenotypic methods depend on growth temperature, carbon source assimilation, chlamydospore and hyphal growth production, positive or negative growth on special media and intracellular enzyme production, among others. In this review, some phenotypic methods are presented with a special emphasis on the discrimination of C. dubliniensis and C. albicans. PMID- 16269033 TI - Clinical features of hypodontia and associated dental anomalies: a retrospective study. AB - OBJECTIVE: Hypodontia is the most commonly known developmental dental anomaly in man. This paper aims to investigate the characteristics of tooth absence and associated dental anomalies among the patients attending our paediatric dentistry clinic. SUBJECTS AND METHODS: For this study, 192 patients aged between 1 and 18 years with congenitally missing teeth were selected among the patients who attended the Department of Paediatric Dentistry of Suleyman Demirel University between January 2000 and December 2003. Clinical and radiographic examinations were performed by two dentists in order to detect the missing teeth. RESULTS: We determined 503 congenitally missing teeth in 192 patients (male = 93, female = 99), excluding third molars. Twelve of these teeth were deciduous and 491 were permanent. A higher incidence of missing teeth was observed in girls than boys, in mandibular arch than maxillary and on the right side than left side. However the differences between arches and sides were not statistically significant in both sexes. CONCLUSION: The diagnosis of the hypodontia should be performed as early as possible, thus alternative treatment modalities can be planned and performed in a multidisciplinary team approach, in order to establish an aesthetic and functional dentition in the future. PMID- 16269034 TI - Preliminary results of moisture checker for Mucus in diagnosing dry mouth. AB - OBJECTIVE: To evaluate the ability of Moisture Checker for Mucus (MCM), a device which measures the weight percent of water content in the oral mucosal epithelium, for the diagnosis of dry mouth. METHOD: Forty-three consecutive patients seen at the Dry Mouth Clinic of Tsurumi University were divided into two groups by the diagnostic criteria of hyposalivation defined by the stimulated salivary flow rate < or =10 ml per 10 min. The MCM values in the buccal, tongue and labial mucosa in each group were compared. RESULTS: The value of the normal salivation group was significantly higher when compared with that of the hyposalivation group both in the buccal and tongue mucosa, P = 0.01 and P = 0.046, respectively. CONCLUSION: Moisture Checker for Mucus has the potential to be a useful device in the screening of hyposalivation. PMID- 16269035 TI - Is the excessive inhibition of matrix metalloproteinases (MMPs) by potent synthetic MMP inhibitors (MMPIs) desirable in periodontitis and other inflammatory diseases? That is: 'Leaky' MMPIs vs excessively efficient drugs. PMID- 16269037 TI - Concurrent or sequential liver and kidney transplantation in children with primary hyperoxaluria type 1? PMID- 16269038 TI - Can non-invasive methodology predict rejection and either dictate or obviate the need for an endomyocardial biopsy in pediatric heart transplant recipients? PMID- 16269039 TI - Transplantation proteomics. AB - The field of proteomics is developing at a rapid pace in the post-genome era. Translational proteomics investigations aim to apply a combination of established methods and new technologies to learn about protein expression profiles predictive of clinical events, therapeutic response, and underlying mechanisms. However, in contrast to genetic studies and in parallel with gene expression studies, the dynamic nature of the proteome in conjunction with the challenges of accounting for post-translational modifications requires the translational proteomics investigator to understand the strengths and limitations of proteomics approaches. In this review, we provide an overview of proteomics approaches and techniques, and proteomics informatics for clinical transplantation investigators. We also review recent publications pertaining to transplantation proteomics, and discuss the implications and utility of urine proteomics for non invasive investigation of transplant outcomes. PMID- 16269041 TI - Cardiac function in children post-orthotopic liver transplantation: echocardiographic parameters and biochemical markers of subclinical cardiovascular damage. AB - Tacrolimus and cyclosporin A (CsA), the mainstay of preventive therapy for solid organ rejection, may cause various side-effects, such as hypertension and nephrotoxicity. Furthermore, tacrolimus is associated with cardiac hypertrophy. In the immediate post-transplant period, both drugs raise the levels of Endothelin-1 (ET), a potent vasoconstrictor; and of B-type Natriuretic Peptide (BNP), a sensitive marker of left ventricular volume overload, which may precede echocardiographic changes of cardiac dysfunction. The aim of the study was to investigate the presence of cardiac damage, by echocardiography and by the biochemical markers BNP and ET, in post-orthotopic liver transplantation (OLT) children, receiving long-term immunosuppressive therapy. ET (ELISA) and BNP (RIA) were measured in plasma of 18 children, post-OLT and 18 healthy controls. Children post-OLT were echocardiographically assessed for left ventricular mass (interventricular septum and posterior wall dimensions), systolic function (ejection fraction, fractional shortening) and diastolic parameters (mitral valve E and A waves, deceleration time, isovolumic relaxation time). None of the post transplant recipients had a history or physical examination consistent with cardiac disease and all recipients were normotensive. Echocardiography revealed no systolic or diastolic dysfunction in any of the recipients. The mean ET and BNP levels tended to be higher among children post-liver transplant, compared with healthy controls (ET: 4.22 +/- 5.35 pg/mL vs. 2.1 +/- 2.0 pg/mL; BNP: 7.05 +/- 4.4 pg/mL vs. 5.87 +/- 2.0 pg/mL, respectively, mean +/- s.d.) although differences did not reach statistical significance. Three children (17%) had elevated BNP and/or ET levels. A strong correlation was observed between ET and BNP levels in post-OLT children (r = 0.79, p < or = 0.05). No correlation was found between ET or BNP levels and echocardiographic findings. In children receiving long-term immunosuppressive therapy post-OLT, although cardiac function is grossly preserved, ET and BNP levels tend to be higher than in healthy, age matched children. Thus, elevated levels of BNP and/or ET may identify patients with early cardiac damage. PMID- 16269042 TI - Outcome of pediatric liver transplant recipients in Turkey: single center experience. AB - To summarize the evolution of the pediatric liver transplant program in a developing country. Between April 1997, and September 2003, 32 cadaveric (CD) and 35 living donor (LD) liver transplantations were performed on 61 children (median age 3.8 yr, range 0.5-16) at Ege University Organ Transplantation and Research Center. The patient's charts were reviewed retrospectively. The outcome of patient and graft survival was analyzed and the incidence of graft loss, complications and rejections was calculated. Indications for liver transplantation were metabolic liver disease (n = 17), biliary atresia (n = 14), viral hepatitis (n = 4), autoimmune hepatitis (n = 6), cryptogenic cirrhosis (n = 11), fulminant liver failure (n = 5) and others (n = 5). Seven of 61 children with chronic liver disease had hepatocellular carcinoma concomitantly. Median pediatric end-stage liver disease score was 23 (range 1-54). Seven children (11.4%) were UNOS status I, 44 (72%) were UNOS status II and 10 (16.6%) were UNOS status III. The median follow-up of the study population was 3.6 yr (range 0.5 6). Actuarial patient survival rates at 1, 2, 3 and 4 yr were 86, 86, 71.3 and 65% in the CD group vs. 80, 76, 67 and 67% in the LR group, respectively (p = NS). Patients listed as UNOS status 1 had lower survival rates than patients listed as UNOS status 2 and 3 (p < 0.05). The mortality rate was 26.2%. Graft survival rates were 81, 81, 75 and 64% at 1, 2, 3 and 4-yr respectively. Six patients (9%) underwent retransplantation. The main complications were infections (64.7%) and surgical complications (43.2%) (including biliary complication, vascular problems, postoperative bleeding, small for size and large for size). The incidence of acute cellular rejection was 39.3%, whereas chronic rejection was 7.4%. The result of liver transplantation in Turkish children was slightly inferior to those reported for North American and European children. However, an important characteristic of these patients that distinguishes them from Europe and North America is that most were UNOS status IIa and UNOS status I (44%). Despite technical and medical progress, infectious and biliary problems have continued to be an important cause of mortality in these patients. PMID- 16269043 TI - Fingerprick blood samples can be used to accurately measure tacrolimus levels by tandem mass spectrometry. AB - Regular monitoring of tacrolimus levels is an essential component of post transplantation follow up in children receiving this drug. We have developed a high performance liquid chromatography tandem mass spectrometry (HPLC T-MS) methodology which measures whole blood tacrolimus levels using a 10 microL sample, thus allowing the use of fingerprick blood sampling. The aim of this study was to determine the degree of relationship between fingerprick and venous tacrolimus levels measured by HPLC T-MS and venous levels measured using the Abbott IMx tacrolimus II assay (IMx). This has not previously been investigated. Blood samples were collected from children on five separate occasions. In addition to the routine venous IMx sample, a further venous and fingerprick sample were collected for tacrolimus level measurement by HPLC T-MS. These were mailed to the central laboratory. One hundred and seventy-two sets of triplicate samples were collected from 36 children (33 kidney, two kidney-pancreas and one kidney-heart). Linear regression analysis showed highly significant relationships between HPLC T-MS venous and IMx venous levels (r(2) = 0.83), HPLC T-MS fingerprick and HPLC T-MS venous levels (r(2) = 0.85) and HPLC T-MS fingerprick and IMx venous levels (r(2) = 0.71) (all p < 0.0001). Bland Altman analysis showed a small, although statistically significant difference between measured values, fingerprick HPLC T-MS levels being lower than venous IMx levels, the mean difference being 0.58 ng/mL (95% CI 0.26-0.91, t = 3.54, d.f. = 168, p = 0.0005 one sample t-test). Precise conversion between the two techniques could be achieved using the regression formula; IMx venous level = 0.751 + 0.978. HPLC T MS fingerprick level. There is a strong significant relationship between fingerprick blood tacrolimus levels measured by HPLC T-MS and venous blood levels measured by IMx, allowing the two to be used interchangeably for routine clinical purposes. PMID- 16269044 TI - Cytomegalovirus infection after allogeneic stem cell transplant in children. AB - The aim of this study was to examine the frequency and the course of cytomegalovirus (CMV) infection and CMV disease in a group of pediatric and adolescent patients after allogeneic stem cell transplantation. Patients were treated according to a protocol including prophylactic high-dose acyclovir in combination with preemptive administration of ganciclovir, based on weekly examinations of CMV antigenemia. A total of 110 consecutively transplanted patients, with a mean age of 9 yr (range 0-20) were treated according to the protocol. All patients were transplanted between March 1993 and January 2000 at the only Danish allotransplantation center. CMV infection occurred in 21.8% (24 of 110) of the patients. Three patients [12.5% (3/24)] developed CMV disease, all with pneumonitis and one with gastrointestinal disease as well. Mean time of disease onset was day +58. Treatment with ganciclovir was in general well tolerated. Late onset CMV disease was not documented. Multivariate analysis revealed that the use of unrelated donor transplants was significantly associated with an increased risk for CMV infection [hazard ratio (HR) 2.90, p = 0.03] and CMV infection was found to be a risk factor for transplant related mortality before day +100 (HR 10.70, p = 0.0015). Although high-dose acyclovir in combination with antigenemia based preemptive treatment with ganciclovir resulted in a low incidence of CMV disease in pediatric and adolescent patients, CMV infection was a significant risk factor in stem cell transplantation with unrelated donors. PMID- 16269045 TI - Long-term results of basiliximab induction immunosuppression in pediatric liver transplant recipients. AB - It has been shown that an induction therapy with the monoclonal anti-interleukin 2 receptor antibody basiliximab (Simulect) is capable to reduce the incidence of acute graft rejection in adult and pediatric liver transplantation (Ltx). However, data on long-term results using basiliximab in children post-Ltx are still pending. Therefore, the objective of our study was to report on the long term results of basiliximab induction therapy in pediatric liver transplant recipients. A total of 54 children received two single doses of basiliximab in addition to cyclosporine and prednisolone following Ltx. We analyzed the incidence of acute and chronic graft rejection that of post-transplant lymphoproliferative disease (PTLD), and patient and graft survival. The follow-up was 22-46 months. The historical control group (matched controls) consisted of 54 patients treated with a cyclosporine and prednisolone dual therapy. Patient survival was 53 of 54 in the treatment group and 51 of 54 in the controls. One patient was retransplanted in the treatment group vs. three patients in the control group. The incidence of acute graft rejection was 16.6% compared with 53.7% in the control group (p < 0.001), that of chronic rejection was comparable in both groups (one of 54 vs. one of 54). The incidence of steroid resistant rejection was four of 54 vs. six of 54 that of PTLD were one of 54 vs. zero of 54. There were no adverse effects observed, which could be related to the antibody treatment. We conclude that basiliximab provides safe and effective induction immunosuppression in pediatric liver graft recipients. Short- and even long-term results are excellent. PMID- 16269046 TI - Prospective monitoring of lipid profiles in children receiving pravastatin preemptively after renal transplantation. AB - Hyperlipidemia is common after renal transplantation (Tx) and contributes to the increased cardiovascular morbidity seen in the post-transplant period. Limited data are available on the utility of the statins in children after renal Tx. This 12-month prospective study was undertaken to determine the efficacy of pravastatin in reducing dyslipidemia after renal Tx in children and to determine predictors of dyslipidemia after Tx. From August 2001 to April 2004, all 17 newly transplanted pediatric renal transplant recipients at our center were preemptively treated with pravastatin from the immediate post-transplant period. Fasting lipid profiles were obtained at 1, 3, 6 and 12 months after Tx. Trends in the lipid profile were analyzed using the repeated measures general linear model (GLM). A historical cohort of pediatric renal-transplant recipients not treated with pravastatin was used as the control population. The mixed effects GLM was used for multivariable logistic regression analyses to determine the independent effect of age, pretransplant cholesterol (Chol), body mass index (BMI), creatinine clearance (CrCl), and corticosteroid and tacrolimus doses on the development of dyslipidemia. The mean age of the children at Tx was 8.7 yr. The GLM analysis showed that with time, there was a significant decline in the total Chol, serum triglyceride (TG), LDL and also HDL-Chol (p-value <0.05 for each). Compared with the controls, the mean serum Chol was lower at all time points post transplant in the treated patients. However, despite treatment, the prevalence of hypercholesterolemia increased from 31% pretransplant to 53% at 1-month, but declined thereafter to 6% at 3 and 6 months and 0% at 1 yr. Multivariable regression analyses showed the prednisone dose, pretransplant Chol and age to be the most important risk factors for the development of dyslipidemia. No child developed complications related to therapy. In summary, pravastatin is safe in the post-transplant period in children and reduces serum Chol, LDL-Chol and TG. An unexpected finding in our study was the decline in HDL-Chol after Tx. Whether the preemptive use of the statins will result in lower cardiovascular morbidity, especially considering the concomitant reduction in HDL-Chol remains to be determined. PMID- 16269047 TI - Better renal function with enhanced immunosuppression and protocol biopsies after kidney transplantation in children. AB - Subclinical rejection may be associated with decreased graft function after renal transplantation (Tx). Detection by protocol biopsies and treatment could thus be important for the long-term prognosis. We have earlier discovered that glomerular filtration rate (GFR) declined in young children during the first 18 months. Consequently, we slightly enhanced and individualized each patient's immunosuppression. This was a retrospective study of 59 pediatric renal Tx patients between 1995 and 2001. The 35 historical controls received triple therapy of azathioprine, methylprednisolone and cyclosporine. GFR was measured by protocol at discharge, 6 and 18 months, and a core biopsy was obtained at 18 months. The 24 study patients in addition received basiliximab, had GFR measured at 3 and 12 months, and a biopsy taken at 3 months. Based on histology and function, immunosuppression was individually adjusted. The groups were compared for GFR and histology at 18 months after Tx. There were less acute rejection episodes in the study group (0.38 vs. 1.23 per patient) and serum creatinine concentrations were lower. Subclinical rejection was detected and treated in 39% at 3 months. There were more chronic changes in the control (47%) than in the study group (29%) at 18 months. GFR was significantly higher in the study group at 18 months (87 vs. 68 mL/min/1.73 m(2)), most remarkably in patients < or =2 yr of age (99 vs. 68 mL/min/1.73 m(2)). Detection of subclinical rejection and slightly enhanced and individualized immunosuppression improved GFR 18 months after renal Tx, especially in the youngest patients. PMID- 16269048 TI - Determinants of graft survival in pediatric and adolescent live donor kidney transplant recipients: a single center experience. AB - To study the independent determinants of graft survival among pediatric and adolescent live donor kidney transplant recipients. Between March 1976 and March 2004, 1600 live donor kidney transplants were carried out in our center. Of them 284 were 20 yr old or younger (mean age 13.1 yr, ranging from 5 to 20 yr). Evaluation of the possible variables that may affect graft survival were carried out using univariate and multivariate analyses. Studied factors included age, gender, relation between donor and recipient, original kidney disease, ABO blood group, pretransplant blood transfusion, human leukocyte antigen (HLA) matching, pretransplant dialysis, height standard deviation score (SDS), pretransplant hypertension, cold ischemia time, number of renal arteries, ureteral anastomosis, time to diuresis, time of transplantation, occurrence of acute tubular necrosis (ATN), primary and secondary immunosuppression, total dose of steroids in the first 3 months, development of acute rejection and post-transplant hypertension. Using univariate analysis, the significant predictors for graft survival were HLA matching, type of primary urinary recontinuity, time to diuresis, ATN, acute rejection and post-transplant hypertension. The multivariate analysis restricted the significance to acute rejection and post-transplant hypertension. The independent determinants of graft survival in live-donor pediatric and adolescent renal transplant recipients are acute rejection and post-transplant hypertension. PMID- 16269049 TI - Pediatric obesity at renal transplantation: a single center experience. AB - Obesity is a major issue affecting health care delivery. While studies have been done in adults with end-stage renal disease, similar studies are lacking in pediatric patients with this disease. We retrospectively analyzed our renal transplant database from 1978 to 2002, to identify prevalence and predisposing factors to obesity in a pediatric end-stage renal disease population. Obesity, particularly in younger individuals, was found to be prevalent at transplantation. PMID- 16269050 TI - Serial signal-averaged electrocardiography in children after cardiac transplantation. AB - In patients with myocytolysis detected in endomyocardial biopsy, there is a tendency towards a shift of ventricular electrical axes from normal to strain pattern on surface ECG. Their 12-lead signal-averaged electrocardiogram (SAECG) show a significant increase in filtered QRS duration (QRSD) compared with those with specimens without myocytolysis. Late potentials were generally found more frequently in individual SAECG leads than in the vector magnitude. An increase in filtered QRSD and the presence of late potentials compared with a baseline study emphasizes the need for endomyocardial biopsy. Consequently, no change in these parameters may preclude the indication for endomyocardial biopsy. PMID- 16269051 TI - Pharmacokinetics of enteric-coated mycophenolate sodium in stable pediatric renal transplant recipients. AB - This study aims to characterize the pharmacokinetics of mycophenolic acid (MPA) and its glucuronide metabolite (mycophenolic acid 7-O-glucuronide, MPAG) following single oral administration of enteric-coated mycophenolate sodium (EC MPS, myfortic) at an approximate dose level of 450 mg/m(2) body surface area (BSA) to 25 stable renal transplant recipients (aged 5-16 yr), and to evaluate the safety and tolerability of EC-MPS in this pediatric population. Patients had been maintained on a cyclosporine emulsion, Neoral-based immunosuppressive regimen for at least 3 months and had received their first or second renal transplant more than 6 months prior to entry into the study. After a brief lag phase (t(lag) 0.75 h), MPA was rapidly absorbed (t(max) 2.5 h) and rapidly converted to MPAG (t(max) 3.25 h), with relatively high plasma concentrations of MPAG (C(max) 67.7 microg/mL) compared with MPA (C(max) 36.3 microg/mL). The elimination half-life for MPAG was slightly longer than for MPA (approximately 13 h vs. 8.5 h), and the apparent oral clearance of MPA was approximately 0.2 L/h/kg. The pharmacokinetics of MPA or MPAG were not affected by age, body weight or BSA, within the study population. The pharmacokinetic results for pediatric patients are comparable with those obtained previously in adults, although exposure based on AUC(0-infinity) was approximately 23% higher, and this finding may be a result of dosing on the basis of BSA, rather than body weight. The recommended dose of EC-MPS in pediatric patients is 400-450 mg/m(2) twice daily or, alternatively, approximately 10-14 mg/kg twice daily when used in combination with cyclosporine microemulsion. PMID- 16269052 TI - Gastric antral vascular ectasia in 2-yr-old girl undergoing unrelated cord blood stem cell transplantation. AB - Gastrointestinal bleeding is a common complication after hematopoietic stem cell transplantation (HSCT) and is often related to acute graft-vs.-host disease (aGVHD). Gastric antral vascular ectasia (GAVE), recently recognized as a complication after HSCT, is a rare cause of severe gastrointestinal bleeding, which has only been reported in adult patients so far. We report a 2-yr-old girl who developed GAVE after unrelated cord blood stem cell transplantation (CBSCT) as treatment of intractable Epstein-Barr virus-associated hemophagocytic lymphohistiocytosis (EBV-HLH). Her conditioning regimen for CBSCT consisted of etoposide, busulfan, and cyclophosphamide. She was doing well after CBSCT without recurrence and developed only grade I aGVHD. She suddenly developed coffee ground emesis, tarry stools and severe anemia 76 days after CBSCT. As antacids were ineffective, esophagogastroduodenoscopy was performed and revealed GAVE on day 97. Endoscopic coagulation therapy was performed twice; subsequently, she needed no further transfusions and there was no clinical recurrence of GAVE. PMID- 16269053 TI - Treatment of acute tacrolimus whole-blood elevation with phenobarbital in the pediatric liver transplant recipient. AB - The toxicities associated with the chronic use of tacrolimus are well described in the literature; however, little is known about the management during an acute overdose. Phenobarbital is a long-acting barbiturate metabolized in the liver by the cytochrome p450 3a4 system. It is known to enhance the rate of metabolism of itself and the clearance of drugs metabolized by p450 3a4. Because tacrolimus is a substrate of this particular isoenzyme, phenobarbital can be considered a potential option when rapid decreases in tacrolimus whole-blood levels are desired. We hereby report our experience using intravenous phenobarbital in the management of two infants with acute elevations in their tacrolimus whole-blood concentration following liver transplantation. Phenobarbital, through its up regulation of hepatic cytochrome p450 system increases the elimination of whole blood tacrolimus concentration in acute overdose situations. PMID- 16269054 TI - Three consecutive related bone marrow transplants for juvenile myelomonocytic leukaemia. AB - Allogeneic haematopoietic stem cell transplantation (allo-HSCT) is the only cure for juvenile myelomonocytic leukaemia (JMML), but relapse remains the major cause of failure. A second transplant may be considered a way to induce the graft vs. leukaemia effect in patients who relapse after their first HSCT. We describe a 7 month-old girl with JMML who relapsed after a first, related allo-HSCT, and who again relapsed 8 months after the second transplant, despite discontinuation of immusuppressive therapy. She underwent a third allogeneic transplant from another related donor. At the time of this report the patient is in complete remission 26 months after the third transplant. We suggest that a third allo-HSCT may be taken into consideration for JMML patients who experience relapse, even after two previous transplants. PMID- 16269055 TI - Acute renal failure in a pediatric kidney allograft recipient treated with intravenous immunoglobulin for parvovirus B19 induced pure red cell aplasia. AB - Infection with parvovirus B19 (PV-B19) after solid organ transplantation may cause pure red cell aplasia (PRCA). Intravenous immunoglobulin (IVIg) may be of benefit in clearing the infection. Acute renal failure is a known adverse effect of IVIg administration. A 14-yr-old male received a cadaveric renal transplant. Three weeks after surgery he developed symptomatic anemia (hemoglobin 4.5 g/dL, reticulocyte count 0.2%). Anti-PV-B19 IgM and IgG titers, which had been negative pretransplant, were positive. He received two IVIg infusions as treatment for the PV-B19 infection. Four days after the IVIg infusions he developed non-oliguric acute renal failure (ARF) with a rise in serum creatinine from 1 to 1.8 mg/dL. Allograft biopsy showed changes consistent with an osmotic load. Anemia and the renal failure resolved after transfusions and IVIg. PV-B19 infection in immunosuppressed transplant recipients is associated with significant morbidity and may respond to IVIg therapy. High sucrose IVIg preparations may be associated with renal failure in renal allograft recipients. Adding PV-B19 testing of the donor and recipient to the standard pretransplant evaluation may be beneficial in diagnosing and managing a potential infection. If IVIg is to be used it may be safer to use a sucrose-free IVIg preparation. PMID- 16269057 TI - Recurrence of hepatic artery thrombosis following acute tacrolimus overdose in pediatric liver transplant recipient. AB - Acute overdose of tacrolimus appears to cause no or minimal adverse clinical consequences. We encountered a pediatric case who underwent liver transplantation associated with hepatic artery thrombosis (HAT), which recurred following acute tacrolimus overdose. A 10-month-old girl underwent living-related liver transplantation because of biliary atresia. To reconstruct the hepatic artery, the right gastroepiploic artery of the donor was interposed between the right hepatic artery of the recipient (2.5 mm in diameter) and the left hepatic graft artery (1 mm in diameter) under microscopy. On postoperative day 4, Doppler ultrasonography showed a remarkable reduction in hepatic arterial flow, which was consistent with HAT. The patient underwent immediate hepatic arteriography and balloon angioplasty. The stenotic sites were dilated by the procedure. Tacrolimus was infused intravenously after transplantation and the infusion rate was adjusted to achieve a target concentration of 18-22 ng/mL, which remained stable until the morning of day 6. An unexpectedly high blood concentration of tacrolimus (57.4 ng/mL) was detected at 6:00 PM on day 6, and tacrolimus was discontinued at 9:00 PM; however, the tacrolimus level reached 119.5 ng/mL at 0:00 h on day 7. While the concentration decreased to 55.2 ng/mL on the morning of day 7, the hepatic arterial flow could not be observed by Doppler ultrasonography. Emergent hepatic arteriography showed stenosis of the artery at the proximal site of the anastomosis. Balloon angioplasty was again performed and the stenotic site was successfully dilated. High level of tacrolimus exposure to the hepatic artery with injured endothelium by preceding angioplasty may have been related to the recurrence of HAT in the present case. PMID- 16269056 TI - Hepatoblastoma in a child with progressive familial intrahepatic cholestasis. AB - End-stage liver cirrhosis because of metabolic or infectious diseases predisposes to hepatic malignancies like hepatocellular carcinoma. We report the first case of hepatoblastoma incidentally detected in the explanted liver of a 2-yr-old child undergoing liver transplantation for cirrhosis because of progressive familial intrahepatic cholestasis (PFIC). The diagnosis was difficult to obtain. The hepatoblastoma was not seen on ultrasound examination of the cirrhotic liver. As we could confirm retrospectively, alpha fetoprotein (AFP) was found elevated prior to transplantation. Two years after successful transplantation, there are no signs of malignancy detectable by clinical and radiological methods. We conclude from this case that PFIC may induce hepatoblastoma and that children with liver cirrhosis should undergo routine screening of serum AFP concentration. PMID- 16269058 TI - Gene expression in skeletal tissues: application of laser capture microdissection. AB - Tissue differentiation is based on the expression of transcription factors, receptors for cytokines, and nuclear receptors that regulate a specific phenotype. The purpose of this study was to select cells from various skeletal tissues in order to analyse differential gene expression of cells in the native environment in vivo. It is a difficult task to obtain cells from skeletal tissues, such as cartilage, periost, bone and muscle, that are structured together and do not exist as individual organs. We used laser capture microdissection which permits the selection and isolation of individual cells from tissue sections. The RNA isolated from these tissues was used for reverse transcriptase-polymerase chain reactions for molecular analysis. We analysed the expression of transcription factors (cFOS, cbfa1, MyoD), receptors for cytokines, nuclear receptors, alkaline phosphatase and the structural proteins osteocalcin and collagen II. The results obtained demonstrate differential patterns of gene expression according to the tissue arrangement in their native in vivo environment, with reliable interpretation of the functions of the analysed genes in the context of intact skeletal tissue physiology. PMID- 16269059 TI - Fully automated intensity compensation for confocal microscopic images. AB - One well-recognized problem in three-dimensional (3D) confocal microscopic images is that the intensities in deeper slices are generally weaker than those in shallower slices. The loss of intensity with depth hampers both qualitative observation and quantitative measurement of specimens. Two major types of methods exist to compensate for this intensity loss: the first is based on the geometrical optics inside the specimen, and the second applies an empirical parametric intensity decay function (IDF) of depth. A common feature shared by both methods is that they are parameter-dependent. However, for the optics-based method there are as yet no fully automated parameter-setting approaches; and for the IDF method the traditional profile-fitting approach cannot provide proper parameters if the presumed IDF model does not match the experimental intensity depth profile of the 3D image. In this paper, we propose a novel maximum-entropy (ME) approach to fully automated parameter-setting. In principle the ME approach is suitable for any compensation method as long as it is parameter-dependent. The basic assumption is that without intensity loss an ideal 3D image should be generally homogeneous with respect to depth and this axial homogeneity can be represented by the entropy of a normalized intensity-depth profile. Experiments on real confocal images showed that such a profile was consistent with visual evaluation of axial intensity homogeneity and that the ME approach could provide proper parameters for both compensation methods mentioned above. Moreover, for the IDF method, experiments on both real and simulated data showed that the ME approach could provide more precise parameters than with traditional profile fitting. The Appendix provides a proof that under certain conditions the global maximization of the profile-entropy is guaranteed. PMID- 16269060 TI - Systematic evaluation of FRAP experiments performed in a confocal laser scanning microscope. AB - The diffusion coefficient as well as the dimensionality of the diffusion process can be determined by straightforward and facile data analysis, when fluorescence recovery after photobleaching (FRAP) is measured as a function of time and space by means of confocal laser scanning microscopy. Experiments representing one dimensional diffusion from a plane source or two-dimensional diffusion from a line source are readily realized. In the data analysis, the deviations of the actual initial conditions from ideal models are consistently taken into account, so that no calibration measurements are needed. The method is applied to FRAP experiments on solutions of Rhodamine B in glycerol and aqueous suspensions of polymethyl methacrylate microspheres. PMID- 16269061 TI - Studies on the influence of power ultrasound on dye penetration in leather dyeing using photomicrographic analysis. AB - The use of power ultrasound in enhancing diffusion rate in various chemical as well as physical processes is gaining in importance. The influence of power ultrasound in the leather dyeing process on enhancing the penetration of dye through the leather matrix was studied. The penetration of dye through a leather cross-section for a given time in the presence and absence of an ultrasonic field (33 kHz, 150 W) was studied by photomicrographic analysis using a stereomicroscope. Different types of black dyes, such as Acid black 1, Metal complex black 194 and Direct black 155, were used for dyeing leather in the present study. Photomicrographic analysis of a cross-section of dyed leather indicated better penetration of dyes through the leather matrix with the use of ultrasound than without it. Therefore, the results indicate that ultrasound helps to improve the diffusion of dye and to reduce diffusional resistance in the leather dyeing process. PMID- 16269062 TI - The characterization of low-angle boundaries by EBSD. AB - A method of accurately measuring misorientations by electron backscatter diffraction (EBSD), which is an extension of that proposed by Wilkinson and based on the comparison of diffraction patterns, is described. The method has been applied to linescans, and found to improve the angular resolution by a factor of more than 30. The consequent improvement in determining misorientation axes is also analysed. Small changes of orientation very close to some low-angle boundaries were investigated and found to be artefacts of the analysis. Measurements of the area from which diffraction patterns are generated show this to be much larger than the effective spatial resolution of EBSD, and it is concluded that this may be a limiting factor in the use of EBSD for microstructural characterization. PMID- 16269063 TI - Improved visualization and quantitative analysis of fluorescent membrane sterol in polarized hepatic cells. AB - Dehydroergosterol is a natural yeast sterol which has recently been employed for direct observation of intracellular sterol transport by UV microscopy. Here, methods are described for improved visualization and quantification of dehydroergosterol in the membranes of polarized HepG2 cells. Using a new online assay, it is shown that dehydroergosterol derived from a cyclodextrin complex inserted into the plasma membrane with a half time of t(1/2) approximately 34 s. Based on a detailed bleaching analysis of dehydroergosterol, slightly different bleaching rates for dehydroergosterol in the basolateral and canalicular membrane were found, indicating different fluorophore environments. Bleaching correction in concert with 3D imaging allows for detection of dehydroergosterol enrichment in microvilli of the canalicular membrane forming the biliary canaliculus. Evidence is provided that some dehydroergosterol accumulating in a subapical compartment or apical recycling compartment can rapidly (t(1/2) approximately 2 min) exchange in vesicles towards the biliary canaliculus while the majority of dehydroergosterol does not redistribute from this compartment. The rapidly exchanging pool resembles only a small portion of the total subapical compartment or apical recycling compartment-associated dehydroergosterol (about 15-30%). Kinetic modelling supports the theory that the subapical compartment or apical recycling compartment to biliary canaliculus transport pathway for sterol is unidirectional. This pathway might be important for rapid biliary transport of free sterol produced by hydrolysis of cholesteryl esters derived from high density lipoprotein. PMID- 16269064 TI - Direct preparation of particles from liquid suspension for ESEM or SEM analysis. AB - A simplified method for the preparation of particles from liquid suspensions has been developed. Particles are deposited directly on carbon planchets for rapid analysis by environmental scanning electron microscopy or by conventional scanning electron microscopy after an additional drying step. This is accomplished by filtering the liquid through thin carbon planchets. Three different grades of graphite were investigated for their suitability as the source material for these planchets. The high quality isomolded graphite is recommended for the filtration and direct observation of particles by electron microscopy. This technique is demonstrated for particles in hydraulic fluid and aquatic suspended particulate material from a natural water source. PMID- 16269065 TI - In vivo assessment of emphysema in mice by high resolution X-ray microtomography. AB - High resolution X-ray microtomography (micro-CT) was used for the detection of emphysema in live mice. Emphysema was induced in C57BL/6 J mice by intratracheal instillation of different amounts of porcine pancreatic elastase. This emphysema could be clearly detected by micro-CT seven weeks post-treatment: analysis of the whole data set of virtual cross-sections showed the presence of a dose-dependent level of emphysema. PMID- 16269066 TI - Hypospadias and maternal exposures to cigarette smoke. AB - The few previous studies of hypospadias and smoking have suggested either no association or a reduced risk. This study, which uses data from the National Birth Defects Prevention Study, a multi-state, population-based case-control study, includes data on males born with severe hypospadias (i.e. the urethra opens at the penile shaft, scrotum or perineum) from 1997 to 2000. Non-malformed, liveborn male controls were selected randomly from birth certificates or from birth hospitals. Maternal interviews were completed by telephone with 453 case mothers and 1267 control mothers. Maternal smoking was not associated with hypospadias risk. For example, during the third month of pregnancy, smoking < 0.5 pack/day had an odds ratio (OR) of 1.1 [95% CI 0.6, 1.9]; 0.5 pack/day, 0.6 [0.4, 1.1]; and > or = 1 pack/day, 0.8 [0.4, 1.6]. Exposure to any secondhand smoke at home during the third month of pregnancy showed an OR of 0.6 [95% CI 0.4, 1.0], and exposure at work or school, an OR of 0.7 [0.5, 1.1]. Similar risks were observed for other months during the periconceptional period, and adjustment for several potential confounders did not substantially alter results. This analysis does not confirm a recent report suggesting that maternal smoking is associated with a reduced risk of having offspring with hypospadias. PMID- 16269068 TI - Population stress and the Swedish sex ratio. AB - Well-developed theory implies that the human secondary sex ratio moves inversely over time with the level of anxiety and depression in the population. Few tests of this hypothesis, however, appear in the voluminous literature concerned with the sex ratio. These tests, moreover, employ designs that allow only weak inference. We contribute to the literature by applying time-series methods to Swedish data for the 276 months beginning January 1974 to detect a relationship between the sex ratio and defined daily doses of antidepressants and anxiolytics dispensed to women. Consistent with theory, we find the drug variable inversely related to the sex ratio. We argue that the discovered association cannot be attributed to shared trends, cycles, or other forms of autocorrelation in the data, or to the problem of endogeneity that necessarily plagues studies based on samples of individual women and births. Implications include that surveillance systems might monitor dispensing of anxiolytics and antidepressants as a marker for population stress thought to be a risk factor for adverse pregnancy outcomes such as preterm delivery. PMID- 16269069 TI - The impact of maternal stress on pregnancy outcome in a well-educated Caucasian population. AB - The aim of the study was to examine the association between stress and pregnancy outcome after adjustment for possible confounding and mediating variables. A prospective cohort study of 5511 pregnancies was conducted in 2001-03 in the Netherlands. A standardised questionnaire collecting demographics and mental health data was administered at 14 and 30 weeks of pregnancy. Medical data on the pregnancy and delivery were obtained from obstetricians and midwives. The results showed that a high level of perceived stress at 14 weeks of pregnancy increased the risk for delivery of an infant that was small-for-gestational-age (OR = 1.26 [95% CI 1.01, 1.56]), but the association was reduced after adjustment for the possible confounding effects of demographic variables (OR = 1.16 [95% CI 0.92, 1.47]). The results do not support a direct relationship between perceived stress and adverse pregnancy outcome. Demographic variables may explain the association between psychosocial stress and pregnancy outcome to a significant degree. PMID- 16269070 TI - A feeling of well-being accompanied by a period of prosperity and birthweight in Chile: a possible link? AB - The aims of the study were to describe the trend in birthweight from 1985 to 2000 in Chile and ascertain the possible factors that may explain the changes in birthweight during the period. We used time series of birthweight and length at birth of all live births (n > 4,000,000). Multivariable regression analyses were carried out to assess whether the gender of the newborn, maternal education, maternal age and marital status could explain the trend in the total time series and by period. Birthweight adjusted for gestational age was analysed in terms of grams and Z scores. There was an increase in birthweight of 100 g between 1989 and 1994 that coincided with the re-establishment of democracy and economic growth in Chile. The changes in birthweight were not explained by the available independent variables and trends of caesarean section, maternal obesity and post mature deliveries over the period. The birthweight difference between mothers with university education and those with primary education was 0.2 Z scores (equivalent to 110 g) in 1985 and was greatly reduced by 2000 (equivalent to 40 g). A positive sense of social well-being and economic growth may have influenced an increase in birthweight between 1989 and 1994 as socio-economic growth preceded and continued after the period of increasing birthweight. Our study showed a remarkable decrease in the differences in birthweight by educational level not shown in other countries of similar wealth. PMID- 16269071 TI - Promotion of folate for the prevention of neural tube defects: who benefits? AB - Since the publication of randomised trials showing firm evidence of prevention of neural tube defects with periconceptional folic acid, there have been population health promotion programmes to encourage women to take folic acid supplements, and the introduction of voluntary fortification of some foods with folic acid in Australia. In order to evaluate these two strategies, we collected data by self administered questionnaire from a random sample of recently pregnant women in Western Australia between September 1997 and March 2000. Response to health promotion was measured in three ways: (1) knowledge of the association between periconceptional folate and prevention of spina bifida (the 'correct message'); (2) use of periconceptional vitamin supplements of folic acid daily in the periconceptional period; and (3) daily folate intake from fortified foods in the 6 months before pregnancy. We examined the relationship of maternal demographic and behavioural characteristics with these three measures. Overall, 62.3% of women were aware of the correct message before pregnancy, 28.5% reported taking 200 microg or more of folic acid from supplements daily in the periconceptional period and 56.6% of women obtained 100 microg or more of folic acid from fortified foods. Women who first became aware of the correct message during pregnancy or who were unaware of the correct message before or during pregnancy were more likely than women aware before pregnancy to be younger, having their first pregnancy, be single or in a de facto relationship, have no tertiary education, and be a public patient. Similar associations were seen for women taking either no folic acid or < 200 microg of folic acid in supplements daily in the periconceptional period. There were no significant associations between these demographic variables and amount of folate obtained from fortified foods. Women who were unaware of the correct message and did not take folic acid supplements were more likely to have smoked, not to have engaged in exercise, and not to have planned their pregnancy, whereas there was no association with these behavioural characteristics and intake of folate from fortified foods. These results indicate that health promotion strategies have not reached all segments of the target population equally, but there is no such disparity with folate-fortified foods, and they suggest that mandatory fortification of a staple food is likely to reach all women regardless of demographic and behavioural characteristics, and hence provide improved opportunity for prevention of neural tube defects in Australia. PMID- 16269072 TI - Does reducing infant mortality depend on preventing low birthweight? An analysis of temporal trends in the Americas. AB - Low birthweight (LBW) is highly associated with death during infancy, and countries with the highest LBW rates also have the highest infant mortality rates. We compared temporal trends in LBW with both overall and birthweight specific infant mortality in United States, Canada, Argentina, Chile, and Uruguay over two time periods, using cohort and cross-sectional analysis of national population-based vital statistics for 1985-89 and 1995-98. Infant mortality diminished substantially (RR = 0.60-0.80 for the later vs. earlier periods) and to a similar degree in all birthweight categories in all five study countries, despite an increase in LBW in the US and Uruguay, minimal changes in Canada and Argentina, and a decrease in Chile. The strength of the (positive) association between LBW and overall infant mortality diminished over the two time periods (from r(s) = +0.80 to +0.25 and RR per SD increase in LBW rate from 2.13 [2.09, 2.17] to 1.76 [1.74, 1.79]). The proportion of infant deaths occurring among LBW infants was negatively correlated with overall infant mortality in both time periods (r(s) = -0.30 and -0.60, RR = 0.68 [0.67, 0.68] and 0.47 [0.46, 0.47]). Developed and less developed countries in the Americas have succeeded in reducing infant mortality in all birthweight groups despite inconsistent changes in LBW rates, and none has achieved this success primarily by reducing LBW. Although our results are not necessarily generalisable to the least developed countries in South Asia and sub-Saharan Africa, it is likely that all countries can substantially reduce their infant mortality rates by improving the care of infants at normal and low birthweights. PMID- 16269073 TI - Childhood intelligence in relation to adult coronary heart disease and stroke risk: evidence from a Danish birth cohort study. AB - While recent studies have reported an inverse relation between childhood intelligence test scores and all-cause mortality in later life, the link with disease-specific outcomes has been rarely examined. Furthermore, the potential confounding effect of birthweight and childhood social circumstances is unknown. We investigated the relation of childhood intelligence with coronary heart disease (CHD) and stroke risk in a cohort of 6910 men born in 1953 in the Copenhagen area of Denmark. Events were ascertained from 1978 to 2000 using a cause-of-death register and hospital discharge records. There were 150 CHD (19 fatal; 131 non-fatal) and 93 stroke (4 fatal; 89 non-fatal) events during follow up into mid-life. Childhood intelligence was inversely related to CHD with the highest rate apparent in adults with low childhood test scores (HR(lowest vs. highest quartile), 2.70; 95% confidence interval: 1.60, 4.57; P(trend) = 0.0001). After adjustment for paternal social class and birthweight, this association was attenuated only marginally. There was little evidence of a IQ-stroke relationship. The cognitive characteristics captured by IQ testing in the present study, such as communication and problem solving ability, appear to be associated with risk of CHD. Health promotion specialists and clinical practitioners may wish to consider these skills in their interactions with the general public. Replication of these results using studies which hold data on intelligence and socio-economic position across the life course is required. PMID- 16269074 TI - Accuracy of reporting maternal in-hospital diagnoses and intrapartum procedures in Washington State linked birth records. AB - While the impact of maternal morbidities and intrapartum procedures is a common topic in perinatal outcomes research, the accuracy of the reporting of these variables in the large administrative databases (birth certificates, hospital discharges) often utilised for such research is largely unknown. We conducted this study to compare maternal diagnoses and procedures listed on birth certificates, hospital discharge data, and birth certificate and hospital discharge data combined, with those documented in a stratified random sample of hospital medical records of 4541 women delivering liveborn infants in Washington State in 2000. We found that birth certificate and hospital discharge data combined had substantially higher true positive fractions (TPF, proportion of women with a positive medical record assessment who were positive using the administrative databases) than did birth certificate data alone for labour induction (86% vs. 52%), cephalopelvic disproportion (83% vs. 35%), abruptio placentae (85% vs. 68%), and forceps-assisted delivery (89% vs. 55%). For procedures available only in hospital discharge data, TPFs were generally high: episiotomy (85%) and third and fourth degree vaginal lacerations (91%). Except for repeat caesarean section without labour (TPF, 81%), delivery procedures available only in birth certificate data had low TPFs, including augmentation (34%), repeat caesarean section with labour (61%), and vaginal birth after caesarean section (62%). Our data suggest that researchers conducting perinatal epidemiological studies should not rely solely on birth certificate data to detect maternal diagnoses and intrapartum procedures accurately. PMID- 16269075 TI - The use and diagnostic yield of radiology in subjects with longstanding musculoskeletal pain--an eight year follow up. AB - BACKGROUND: Longstanding musculoskeletal pain is common in the general population and associated with frequent use of health care. Plain radiography is a common diagnostic approach in these patients despite knowledge that the use in the investigation of musculoskeletal pain is associated with low diagnostic yield, substantial costs and high radiation exposure. The aim of this study was to assess the use of diagnostic imaging and the proportion of pathological findings with regard to duration and distribution of pain in a cohort from the general population. METHODS: An eight-year longitudinal study based on questionnaires at three occasions and medical records on radiological examinations done in medical care. Thirty subjects were selected from an established population based cohort of 2425 subjects that in 1995 answered a postal survey on pain experience. At baseline there were ten subjects from each of three pain groups; No chronic pain, Chronic regional pain, and Chronic widespread pain (CWP). Those who presented with CWP at two or all three occasions were considered to have a longstanding or re-occurring CWP. In total the thirty subjects underwent 102 radiological examinations during the eight year follow up. RESULTS: There was a non significant (p = 0.10) finding indicating that subjects with chronic pain at baseline (regional or widespread) were examined three times more often than those with no chronic pain. When the indication for the examination was pain, there was a low proportion of positive findings in subjects with longstanding CWP, compared to all others (5.3% vs 28.9%; p = 0.045). On the other hand, in examinations on other indications than pain the proportion of positive findings was high in the CWP group (62.5% vs 14.8%; p = 0.001). CONCLUSION: Radiological examinations had a low diagnostic yield in evaluation of pain in subjects with longstanding/reoccurring CWP. These subjects had on the other hand more often positive findings when examined on other indications than pain. This may indicate that subjects with longstanding/reoccurring CWP are more prone to other diseases. It is a challenge for caregivers, often primary care physicians, to use radiological examinations to the best for their patients. PMID- 16269076 TI - A fully automatable enzymatic method for DNA extraction from plant tissues. AB - BACKGROUND: DNA extraction from plant tissues, unlike DNA isolation from mammalian tissues, remains difficult due to the presence of a rigid cell wall around the plant cells. Currently used methods inevitably require a laborious mechanical grinding step, necessary to disrupt the cell wall for the release of DNA. RESULTS: Using a cocktail of different carbohydrases, a method was developed that enables a complete digestion of the plant cell walls and subsequent DNA release. Optimized conditions for the digestion reaction minimize DNA shearing and digestion, and maximize DNA release from the plant cell. The method gave good results in 125 of the 156 tested species. CONCLUSION: In combination with conventional DNA isolation techniques, the new enzymatic method allows to obtain high-yield, high-molecular weight DNA, which can be used for many applications, including genome characterization by AFLP, RAPD and SSR. Automation of the protocol (from leaf disks to DNA) is possible with existing workstations. PMID- 16269077 TI - maxdLoad2 and maxdBrowse: standards-compliant tools for microarray experimental annotation, data management and dissemination. AB - BACKGROUND: maxdLoad2 is a relational database schema and Java application for microarray experimental annotation and storage. It is compliant with all standards for microarray meta-data capture; including the specification of what data should be recorded, extensive use of standard ontologies and support for data exchange formats. The output from maxdLoad2 is of a form acceptable for submission to the ArrayExpress microarray repository at the European Bioinformatics Institute. maxdBrowse is a PHP web-application that makes contents of maxdLoad2 databases accessible via web-browser, the command-line and web service environments. It thus acts as both a dissemination and data-mining tool. RESULTS: maxdLoad2 presents an easy-to-use interface to an underlying relational database and provides a full complement of facilities for browsing, searching and editing. There is a tree-based visualization of data connectivity and the ability to explore the links between any pair of data elements, irrespective of how many intermediate links lie between them. Its principle novel features are: the flexibility of the meta-data that can be captured, the tools provided for importing data from spreadsheets and other tabular representations, the tools provided for the automatic creation of structured documents, the ability to browse and access the data via web and web-services interfaces. Within maxdLoad2 it is very straightforward to customise the meta-data that is being captured or change the definitions of the meta-data. These meta-data definitions are stored within the database itself allowing client software to connect properly to a modified database without having to be specially configured. The meta-data definitions (configuration file) can also be centralized allowing changes made in response to revisions of standards or terminologies to be propagated to clients without user intervention.maxdBrowse is hosted on a web-server and presents multiple interfaces to the contents of maxd databases. maxdBrowse emulates many of the browse and search features available in the maxdLoad2 application via a web-browser. This allows users who are not familiar with maxdLoad2 to browse and export microarray data from the database for their own analysis. The same browse and search features are also available via command-line and SOAP server interfaces. This both enables scripting of data export for use embedded in data repositories and analysis environments, and allows access to the maxd databases via web-service architectures. CONCLUSION: maxdLoad2 http://www.bioinf.man.ac.uk/microarray/maxd/ and maxdBrowse http://dbk.ch.umist.ac.uk/maxdBrowse are portable and compatible with all common operating systems and major database servers. They provide a powerful, flexible package for annotation of microarray experiments and a convenient dissemination environment. They are available for download and open sourced under the Artistic License. PMID- 16269078 TI - Towards understanding the presence/absence of Human African Trypanosomosis in a focus of Cote d'Ivoire: a spatial analysis of the pathogenic system. AB - BACKGROUND: This study aimed at identifying factors influencing the development of Human African Trypanosomosis (HAT, or sleeping sickness) in the focus of Bonon, located in the mesophile forest of Cote d'Ivoire. A previous study mapping the main daytime activity sites of 96 patients revealed an important disparity between the area south of the town- where all the patients lived- and the area north of the town, apparently free of disease. In order to explain this disparity, we carried out a spatial analysis of the key components of the pathogenic system, i.e. the human host, the tsetse vector and the trypanosomes in their environment using a geographic information system (GIS). RESULTS: This approach at the scale of a HAT focus enabled us to identify spatial patterns which linked to the transmission and the dissemination of this disease. The history of human settlement (with the rural northern area exploited much earlier than the southern one) appears to be a major factor which determines the land use pattern, which itself may account for differences found in vector densities (tsetse were found six times more abundant in the southern rural area than in the northern). Vector density, according to the human and environmental context in which it is found (here an intense mobility between the town of Bonon and the rural areas), may explain the observed spatial differences in HAT prevalence. CONCLUSION: This work demonstrates the role of GIS analyses of key components of the pathogenic system in providing a better understanding of transmission and dissemination of HAT. Moreover, following the identification of the most active transmission areas, and of an area unfavourable to HAT transmission, this study more precisely delineates the boundaries of the Bonon focus. As a follow-up, targeted tsetse control activities starting north of Bonon (with few chances of reinvasion due to very low densities) going south, and additional medical surveys in the south will be proposed to the Ivoirian HAT control program to enhance the control of the disease in this focus. This work also shows the evolution of HAT regarding time and environment, and the methodology used may be able to predict possible sleeping sickness development/extinction in areas with similar history and space organization. PMID- 16269080 TI - Human leukocyte antigen class I, class II, and tumor necrosis factor-alpha polymorphisms in a healthy elder Mexican Mestizo population. AB - BACKGROUND: There is strong evidence that an individual's genetic background is an important predisposing factor to longevity. In the present study we analysed the frequency of HLA class I, class II, as well as the TNF-alpha -308 polymorphism that may be related to an increased life span in Mexican Mestizo healthy elders. RESULTS: HLA typing was performed by polymerase chain reaction sequence specific oligonucleotide (PCR SSO) reverse dot blot. The TNF-alpha -308 polymorphism was assessed by PCR restriction fragment length polymorphism. A significant increased frequency of HLA-DRB1*11 was found in elderly women whereas this allele was not present in elderly males. The TNF2 allele was also increased in the elder group when compared to young controls. The frequencies of the remaining alleles tested were not statistically different among groups. CONCLUSION: These data suggest an ethnicity independent tendency of HLA-DRB1*11 in elder females to increase life span and a possible role of the TNF2 allele with the successful remodelling of senescent immune system. PMID- 16269081 TI - Parametric versus non-parametric statistics in the analysis of randomized trials with non-normally distributed data. AB - BACKGROUND: It has generally been argued that parametric statistics should not be applied to data with non-normal distributions. Empirical research has demonstrated that Mann-Whitney generally has greater power than the t-test unless data are sampled from the normal. In the case of randomized trials, we are typically interested in how an endpoint, such as blood pressure or pain, changes following treatment. Such trials should be analyzed using ANCOVA, rather than t test. The objectives of this study were: a) to compare the relative power of Mann Whitney and ANCOVA; b) to determine whether ANCOVA provides an unbiased estimate for the difference between groups; c) to investigate the distribution of change scores between repeat assessments of a non-normally distributed variable. METHODS: Polynomials were developed to simulate five archetypal non-normal distributions for baseline and post-treatment scores in a randomized trial. Simulation studies compared the power of Mann-Whitney and ANCOVA for analyzing each distribution, varying sample size, correlation and type of treatment effect (ratio or shift). RESULTS: Change between skewed baseline and post-treatment data tended towards a normal distribution. ANCOVA was generally superior to Mann Whitney in most situations, especially where log-transformed data were entered into the model. The estimate of the treatment effect from ANCOVA was not importantly biased. CONCLUSION: ANCOVA is the preferred method of analyzing randomized trials with baseline and post-treatment measures. In certain extreme cases, ANCOVA is less powerful than Mann-Whitney. Notably, in these cases, the estimate of treatment effect provided by ANCOVA is of questionable interpretability. PMID- 16269079 TI - cGMP-independent nitric oxide signaling and regulation of the cell cycle. AB - BACKGROUND: Regulatory functions of nitric oxide (NO*) that bypass the second messenger cGMP are incompletely understood. Here, cGMP-independent effects of NO* on gene expression were globally examined in U937 cells, a human monoblastoid line that constitutively lacks soluble guanylate cyclase. Differentiated U937 cells (>80% in G0/G1) were exposed to S-nitrosoglutathione, a NO* donor, or glutathione alone (control) for 6 h without or with dibutyryl-cAMP (Bt2cAMP), and then harvested to extract total RNA for microarray analysis. Bt2cAMP was used to block signaling attributable to NO*-induced decreases in cAMP. RESULTS: NO* regulated 110 transcripts that annotated disproportionately to the cell cycle and cell proliferation (47/110, 43%) and more frequently than expected contained AU rich, post-transcriptional regulatory elements (ARE). Bt2cAMP regulated 106 genes; cell cycle gene enrichment did not reach significance. Like NO*, Bt2cAMP was associated with ARE-containing transcripts. A comparison of NO* and Bt2cAMP effects showed that NO* regulation of cell cycle genes was independent of its ability to interfere with cAMP signaling. Cell cycle genes induced by NO* annotated to G1/S (7/8) and included E2F1 and p21/Waf1/Cip1; 6 of these 7 were E2F target genes involved in G1/S transition. Repressed genes were G2/M associated (24/27); 8 of 27 were known targets of p21. E2F1 mRNA and protein were increased by NO*, as was E2F1 binding to E2F promoter elements. NO* activated p38 MAPK, stabilizing p21 mRNA (an ARE-containing transcript) and increasing p21 protein; this increased protein binding to CDE/CHR promoter sites of p21 target genes, repressing key G2/M phase genes, and increasing the proportion of cells in G2/M. CONCLUSION: NO* coordinates a highly integrated program of cell cycle arrest that regulates a large number of genes, but does not require signaling through cGMP. In humans, antiproliferative effects of NO* may rely substantially on cGMP-independent mechanisms. Stress kinase signaling and alterations in mRNA stability appear to be major pathways by which NO* regulates the transcriptome. PMID- 16269082 TI - A mathematical model of the euglycemic hyperinsulinemic clamp. AB - BACKGROUND: The Euglycemic Hyperinsulinemic Clamp (EHC) is the most widely used experimental procedure for the determination of insulin sensitivity, and in its usual form the patient is followed under insulinization for two hours. In the present study, sixteen subjects with BMI between 18.5 and 63.6 kg/m(2) were studied by long-duration (five hours) EHC. RESULTS: From the results of this series and from similar reports in the literature it is clear that, in obese subjects, glucose uptake rates continue to increase if the clamp procedure is prolonged beyond the customary 2 hours. A mathematical model of the EHC, incorporating delays, was fitted to the recorded data, and the insulin resistance behaviour of obese subjects was assessed analytically. Obese subjects had significantly less effective suppression of hepatic glucose output and higher pancreatic insulin secretion than lean subjects. Tissue insulin resistance appeared to be higher in the obese group, but this difference did not reach statistical significance. CONCLUSION: The use of a mathematical model allows a greater amount of information to be recovered from clamp data, making it easier to understand the components of insulin resistance in obese vs. normal subjects. PMID- 16269083 TI - The Bradford Hill considerations on causality: a counterfactual perspective. AB - Bradford Hill's considerations published in 1965 had an enormous influence on attempts to separate causal from non-causal explanations of observed associations. These considerations were often applied as a checklist of criteria, although they were by no means intended to be used in this way by Hill himself. Hill, however, avoided defining explicitly what he meant by "causal effect". This paper provides a fresh point of view on Hill's considerations from the perspective of counterfactual causality. I argue that counterfactual arguments strongly contribute to the question of when to apply the Hill considerations. Some of the considerations, however, involve many counterfactuals in a broader causal system, and their heuristic value decreases as the complexity of a system increases; the danger of misapplying them can be high. The impacts of these insights for study design and data analysis are discussed. The key analysis tool to assess the applicability of Hill's considerations is multiple bias modelling (Bayesian methods and Monte Carlo sensitivity analysis); these methods should be used much more frequently. PMID- 16269084 TI - The contribution of leading diseases and risk factors to excess losses of healthy life in Eastern Europe: burden of disease study. AB - BACKGROUND: The East/West gradient in health across Europe has been described often, but not using metrics as comprehensive and comparable as those of the Global Burden of Disease 2000 and Comparative Risk Assessment studies. METHODS: Comparisons are made across 3 epidemiological subregions of the WHO region for Europe--A (very low child and adult mortality), B (low child and low adult mortality) and C (low child and high adult mortality)--with populations in 2000 of 412, 218 and 243 millions respectively, and using the following measures: 1. Probabilities of death by sex and causal group across 7 age intervals; 2. Loss of healthy life (DALYs) to diseases and injuries per thousand population; 3. Loss of healthy life (DALYs) attributable to selected risk factors across 3 age ranges. RESULTS: Absolute differences in mortality are most marked in males and in younger adults, and for deaths from vascular diseases and from injuries. Dominant contributions to east-west differences come from the nutritional/physiological group of risk factors (blood pressure, cholesterol concentration, body mass index, low fruit and vegetable consumption and inactivity) contributing to vascular disease and from the legal drugs--tobacco and alcohol. CONCLUSION: The main requirements for reducing excess health losses in the east of Europe are: 1) favorable shifts in all amenable vascular risk factors (irrespective of their current levels) by population-wide and personal measures; 2) intensified tobacco control; 3) reduced alcohol consumption and injury control strategies (for example, for road traffic injuries). Cost effective strategies are broadly known but local institutional support for them needs strengthening. PMID- 16269085 TI - Disseminated tuberculosis presenting with finger swelling in a patient with tuberculous osteomyelitis: a case report. AB - BACKGROUND: Extrapulmonary manifestations of tuberculosis have become increasingly important in the era of HIV/AIDS. CASE PRESENTATION: We describe a case of tuberculosis (TB) dactylitis in a patient with AIDS who originated from the Ivory Coast. The diagnosis was established by direct visualization of acid fast bacilli on joint fluid and bone biopsy of the proximal phalanx. Imaging of the chest revealed multiple bilateral nodules. Confirmation of the diagnosis was made by isolation of Mycobacterium tuberculosis from sputum and bone cultures. CONCLUSION: Tuberculosis should be considered in patients with unusual soft tissue or skeletal lesions, especially when an immunosuppressive condition is present. Ziehl-Neelsen staining and culture of tissue obtained via surgical biopsy offer the most direct approach to diagnosis. PMID- 16269086 TI - Characterization of the metabolic shift between oxidative and fermentative growth in Saccharomyces cerevisiae by comparative 13C flux analysis. AB - BACKGROUND: One of the most fascinating properties of the biotechnologically important organism Saccharomyces cerevisiae is its ability to perform simultaneous respiration and fermentation at high growth rate even under fully aerobic conditions. In the present work, this Crabtree effect called phenomenon was investigated in detail by comparative 13C metabolic flux analysis of S. cerevisiae growing under purely oxidative, respiro-fermentative and predominantly fermentative conditions. RESULTS: The metabolic shift from oxidative to fermentative growth was accompanied by complex changes of carbon flux throughout the whole central metabolism. This involved a flux redirection from the pentose phosphate pathway (PPP) towards glycolysis, an increased flux through pyruvate carboxylase, the fermentative pathways and malic enzyme, a flux decrease through the TCA cycle, and a partial relocation of alanine biosynthesis from the mitochondrion to the cytosol. S. cerevisiae exhibited a by-pass of pyruvate dehydrogenase in all physiological regimes. During oxidative growth this by-pass was mainly provided via pyruvate decarboxylase, acetaldehyde dehydrogenase, acetyl-CoA synthase and transport of acetyl-CoA into the mitochondrion. During fermentative growth this route, however, was saturated due to limited enzyme capacity. Under these conditions the cells exhibited high carbon flux through a chain of reactions involving pyruvate carboxylase, the oxaloacetate transporter and malic enzyme. During purely oxidative growth the PPP alone was sufficient to completely supply NADPH for anabolism. During fermentation, it provided only 60 % of the required NADPH. CONCLUSION: We conclude that, in order to overcome the limited capacity of pyruvate dehydrogenase, S. cerevisiae possesses different metabolic by-passes to channel carbon into the mitochondrion. This involves the conversion of cytosolic pyruvate either into acetyl CoA or oxaloacetate followed by intercompartmental transport of these metabolites. During oxidative growth mainly the NAD specific isoforms of acetaldehyde dehydrogenase and isocitrate dehydrogenase catalyze the corresponding reactions in S. cerevisiae, whereas NADPH supply under fermentative conditions involves significant contribution of sources other than the PPP such as e. g. NADPH specific acetaldehyde dehydrogenase or isocitrate dehydrogenase. PMID- 16269087 TI - Epidural abscess caused by Streptococcus milleri in a pregnant woman. AB - BACKGROUND: Bacteria in the Streptococcus milleri group (S. anginosus, S. constellatus, and S. intermedius) are associated with bacteremia and abscess formation. While most reports of Streptococcus milleri group (SMG) infection occur in patients with underlying medical conditions, SMG infections during pregnancy have been documented. However, SMG infections in pregnant women are associated with either neonatal or maternal puerperal sepsis. Albeit rare, S. milleri spinal-epidural abscess in pregnancy has been reported, always as a complication of spinal-epidural anesthesia. We report a case of spinal-epidural abscess caused by SMG in a young, pregnant woman without an antecedent history of spinal epidural anesthesia and without any underlying risk factors for invasive streptococcal disease. CASE PRESENTATION: A 25 year old pregnant woman developed neurological symptoms consistent with spinal cord compression at 20 weeks gestation. She underwent emergency laminectomy for decompression and was treated with ceftriaxone 2 gm IV daily for 28 days. She was ambulatory at the time of discharge from the inpatient rehabilitation unit with residual lower extremity weakness. CONCLUSION: To our knowledge, this is the first reported case of a Streptococcus milleri epidural abscess in a healthy, pregnant woman with no history of epidural anesthesia or invasive procedures. This report adds to the body of literature on SMG invasive infections. Treatment of SMG spinal-epidural abscess with neurologic manifestations should include prompt and aggressive surgical decompression coupled with targeted anti-infective therapy. PMID- 16269088 TI - Evaluation of KO-Tab 1-2-3: a wash-resistant 'dip-it-yourself' insecticide formulation for long-lasting treatment of mosquito nets. AB - INTRODUCTION: Insecticide-treated nets (ITN) are an important method of preventing malaria. To remain effective, they need to be re-treated with pyrethroid insecticide at approximately yearly intervals. Systems for re-treating nets in Africa are limited, and the vast majority of nets in use have never been treated or were treated only once. Bayer Environmental Science (BES) has developed a long-lasting formulation 'KO-Tab 1-2-3' which can be applied to the net post-manufacture, under field conditions, and renders the insecticide wash resistant. METHODS: The performance of polyester nets treated with three kinds of BES long-lasting formulations, a conventional ITN (treated with standard KO-Tab) and PermaNet 2.0 were evaluated after washing samples of treated netting up to 30 times using standard WHO procedures. Performance was measured using 'three-minute exposure' and 'median time to knockdown' bioassay tests and by measuring the levels of deltamethrin using high-pressure liquid chromatography. RESULTS: The conventional ITN was largely stripped of deltamethrin within 5-10 washes and insecticidal efficacy in bioassay declined to suboptimal levels. With PermaNet and KO-Tab 1-2-3 the loss of deltamethrin was much slower: insecticide content halved within 20 washes and there was no loss of biological efficacy in three minute exposure bioassays in WHO cylinders even after 30 washes. After 30 washes there remained on the netting 16% (4.4 mg/m2) of the loading dose of KO-Tab 1-2-3 and 28% (18.8 mg/m2) of the loading dose of PermaNet. CONCLUSION: KO-Tab 1-2-3 was confirmed to be a long-lasting insecticide formulation. This finding raises the prospect of conventional polyester nets being converted into long-lasting insecticidal nets through simple dipping in the community or at home. This single development, if widely adopted, could transform the malaria control landscape in Africa and have a major impact on malaria. PMID- 16269089 TI - Primary small cell neuroendocrine carcinoma of the urinary bladder with coexisting high-grade urothelial carcinoma: a case report and a review of the literature. AB - Primary neuroendocrine carcinomas of the urinary bladder are rare. Here, we report a case of an 82-year-old man who presented with hematuria and was found to have an ulcerated lesion in the bladder. A diagnosis of small neuroendocrine cell carcinoma with coexisting minor high-grade urothelial components was rendered. In this report, the clinical, cytological, histological, and immunohistochemical features of this case are described, and a review of the literature about this neoplasm is presented. The differential diagnoses of small cell tumor in urinary bladder washing specimens are discussed. PMID- 16269090 TI - Isolation of human beta-defensin-4 in lung tissue and its increase in lower respiratory tract infection. AB - BACKGROUND: Human beta-defensin-4 (hBD-4), a new member of the beta-defensin family, was discovered by an analysis of the genomic sequence. The objective of this study was to clarify hBD-4 expression in human lung tissue, along with the inducible expression in response to infectious stimuli, localization, and antimicrobial activities of hBD-4 peptides. We also investigated the participation of hBD-4 in chronic lower respiratory tract infections (LRTI) by measuring the concentrations of hBD-4 peptides in human bronchial epithelial lining fluid (ELF). METHODS: The antimicrobial activity of synthetic hBD-4 peptides against E. coli and P. aeruginosa was measured by radial diffusion and colony count assays. We identified hBD-4 in homogenated human lung tissue by reverse-phase high-performance liquid chromatography coupled with a radioimmunoassay (RIA). Localization of hBD-4 was studied through immunohistochemical analysis (IHC). We investigated the effects of lipopolysaccharide (LPS) on hBD-4 expression and its release from small airway epithelial cells (SAEC). We collected ELF from patients with chronic LRTI using bronchoscopic microsampling to measure hBD-4 concentrations by RIA. RESULTS: hBD 4 exhibited salt-sensitive antimicrobial activity against P. aeruginosa. We detected the presence of hBD-4 peptides in human lung tissue. IHC demonstrated the localization of hBD-4-producing cells in bronchial and bronchiolar epithelium. The levels of hBD-4 peptides released from LPS-treated SAECs were higher than those of untreated control cells. ELF hBD-4 was detectable in 4 of 6 patients with chronic LRTI, while the amounts in controls were all below the detectable level. CONCLUSION: This study suggested that hBD-4 plays a significant role in the innate immunity of the lower respiratory tract. PMID- 16269091 TI - RB1 gene mutation up-date, a meta-analysis based on 932 reported mutations available in a searchable database. AB - BACKGROUND: Retinoblastoma, a prototype of hereditary cancer, is the most common intraocular tumour in children and potential cause of blindness from therapeutic eye ablation, second tumours in germ line carrier's survivors, and even death when left untreated. The molecular scanning of RB1 in search of germ line mutations lead to the publication of more than 900 mutations whose knowledge is important for genetic counselling and the characterization of phenotypic genotypic relationships. RESULTS: A searchable database (RBGMdb) has been constructed with 932 published RB1 mutations. The spectrum of these mutations has been analyzed with the following results: 1) the retinoblastoma protein is frequently inactivated by deletions and nonsense mutations while missense mutations are the main inactivating event in most genetic diseases. 2) Near 40% of RB1 gene mutations are recurrent and gather in sixteen hot points, including twelve nonsense, two missense and three splicing mutations. The remainder mutations are scattered along RB1, being most frequent in exons 9, 10, 14, 17, 18, 20, and 23. 3) The analysis of RB1 mutations by country of origin of the patients identifies two groups in which the incidence of nonsense and splicing mutations show differences extremely significant, and suggest the involvement of predisposing ethnic backgrounds. 4) A significant association between late age at diagnosis and splicing mutations in bilateral retinoblastoma patients suggests the occurrence of a delayed-onset genotype. 5) Most of the reported mutations in low-penetrance families fall in three groups: a) Mutations in regulatory sequences at the promoter resulting in low expression of a normal Rb; b) Missense and in-frame deletions affecting non-essential sequence motifs which result in a partial inactivation of Rb functions; c) Splicing mutations leading to the reduction of normal mRNA splicing or to alternative splicing involving either true oncogenic or defective (weak) alleles. CONCLUSION: The analysis of RB1 gene mutations logged in the RBGMdb has shown relevant phenotype-genotype relationships and provided working hypothesis to ascertain mechanisms linking certain mutations to ethnicity, delayed onset of the disease and low-penetrance. Gene profiling of tumors will help to clarify the genetic background linked to ethnicity and variable expressivity or delayed onset phenotypes. PMID- 16269092 TI - Using psilocybin to investigate the relationship between attention, working memory, and the serotonin 1A and 2A receptors. AB - Increasing evidence suggests a link between attention, working memory, serotonin (5-HT), and prefrontal cortex activity. In an attempt to tease out the relationship between these elements, this study tested the effects of the hallucinogenic mixed 5-HT1A/2A receptor agonist psilocybin alone and after pretreatment with the 5-HT2A antagonist ketanserin. Eight healthy human volunteers were tested on a multiple-object tracking task and spatial working memory task under the four conditions: placebo, psilocybin (215 microg/kg), ketanserin (50 mg), and psilocybin and ketanserin. Psilocybin significantly reduced attentional tracking ability, but had no significant effect on spatial working memory, suggesting a functional dissociation between the two tasks. Pretreatment with ketanserin did not attenuate the effect of psilocybin on attentional performance, suggesting a primary involvement of the 5-HT1A receptor in the observed deficit. Based on physiological and pharmacological data, we speculate that this impaired attentional performance may reflect a reduced ability to suppress or ignore distracting stimuli rather than reduced attentional capacity. The clinical relevance of these results is also discussed. PMID- 16269093 TI - Analysis of single-unit responses to emotional scenes in human ventromedial prefrontal cortex. AB - Lesion and functional imaging studies in humans have shown that the ventral and medial prefrontal cortex is critically involved in the processing of emotional stimuli, but both of these methods have limited spatiotemporal resolution. Conversely, neurophysiological studies of emotion in nonhuman primates typically rely on stimuli that do not require elaborate cognitive processing. To begin bridging this gap, we recorded from a total of 267 neurons in the left and right orbital and anterior cingulate cortices of four patients who had chronically implanted depth electrodes for monitoring epilepsy. Peristimulus activity was recorded to standardized, complex visual scenes depicting neutral, pleasant, or aversive content. Recording locations were verified with postoperative magnetic resonance imaging. Using a conservative, multistep statistical evaluation, we found significant responses in 56 neurons; 16 of these were selective for only one emotion class, most often aversive. The findings suggest sparse and widely distributed processing of emotional value in the prefrontal cortex, with a predominance of responses to aversive stimuli. PMID- 16269095 TI - Emerging neurophysiological specialization for letter strings. AB - In adult readers, printed words and other letter strings activate specialized visual functions within 200 msec, as evident from neurophysiological recordings of brain activity. These fast, specialized responses to letter strings are thought to develop through plastic changes in the visual system. However, it is unknown whether this specialization emerges only with the onset of word reading, or represents a precursor of literacy. We compared 6-year-old kindergarten children who could not yet read words to adult readers. Both age groups detected immediate repetitions of visually presented words, pseudowords, symbol strings, and pictures during event-related potential (ERP) mapping. Maps from seven corresponding ERP segments in children and adults were analyzed regarding fast (<250 msec) and slow (>300 msec) specialization for letter strings. Adults reliably differentiated words through increased fast (<150 msec) occipito temporal N1 activity from symbols. Children showed a later, more mid-occipital N1 with marginal word-symbol differences, which were absent in those children with low letter knowledge. Children with high letter knowledge showed some fast sensitivity to letter strings, which was confined to right occipito-temporal sites, unlike the stronger adult N1 specialization. This suggests that a critical degree of early literacy induces some immature, but fast, specialization for letter strings before word reading becomes possible. Children also differentiated words from symbols in later segments through increased right occipito-temporal negativity for words. This slow specialization for letter strings was not modulated by letter knowledge and was absent in adults, possibly reflecting a visual precursor of literacy due to visual familiarity with letter strings. PMID- 16269094 TI - Left auditory cortex and amygdala, but right insula dominance for human laughing and crying. AB - Evidence suggests that in animals their own species-specific communication sounds are processed predominantly in the left hemisphere. In contrast, processing linguistic aspects of human speech involves the left hemisphere, whereas processing some prosodic aspects of speech as well as other not yet well-defined attributes of human voices predominantly involves the right hemisphere. This leaves open the question of hemispheric processing of universal (species specific) human vocalizations that are more directly comparable to animal vocalizations. The present functional magnetic resonance imaging study addresses this question. Twenty subjects listened to human laughing and crying presented either in an original or time-reversed version while performing a pitch-shift detection task to control attention. Time-reversed presentation of these sounds is a suitable auditory control because it does not change the overall spectral content. The auditory cortex, amygdala, and insula in the left hemisphere were more strongly activated by original than by time-reversed laughing and crying. Thus, similar to speech, these nonspeech vocalizations involve predominantly left hemisphere auditory processing. Functional data suggest that this lateralization effect is more likely based on acoustical similarities between speech and laughing or crying than on similarities with respect to communicative functions. Both the original and time-reversed laughing and crying activated more strongly the right insula, which may be compatible with its assumed function in emotional self-awareness. PMID- 16269096 TI - fMRI reveals a common neural substrate of illusory and real contours in V1 after perceptual learning. AB - Perceptual learning involves the specific and relatively permanent modification of perception following a sensory experience. In psychophysical experiments, the specificity of the learning effects to the trained stimulus attributes (e.g., visual field position or stimulus orientation) is often attributed to assumed neural modifications at an early cortical site within the visual processing hierarchy. We directly investigated a neural correlate of perceptual learning in the primary visual cortex using fMRI. Twenty volunteers practiced a curvature discrimination on Kanizsa-type illusory contours in the MR scanner. Practice induced changes in the BOLD response to illusory contours were compared between the pretraining and the posttraining block in those areas of the primary visual cortex (V1) that, in the same session, had been identified to represent real contours at corresponding visual field locations. A retinotopically specific BOLD signal increase to illusory contours was observed as a consequence of the training, possibly signaling the formation of a contour representation, which is necessary for performing the curvature discrimination. The effects of perceptual training were maintained over a period of about 10 months, and they were specific to the trained visual field position. The behavioral specificity of the learning effects supports an involvement of V1 in perceptual learning, and not in unspecific attentional effects. PMID- 16269097 TI - Interaction between syntax processing in language and in music: an ERP Study. AB - The present study investigated simultaneous processing of language and music using visually presented sentences and auditorily presented chord sequences. Music-syntactically regular and irregular chord functions were presented synchronously with syntactically correct or incorrect words, or with words that had either a high or a low semantic cloze probability. Music-syntactically irregular chords elicited an early right anterior negativity (ERAN). Syntactically incorrect words elicited a left anterior negativity (LAN). The LAN was clearly reduced when words were presented simultaneously with music syntactically irregular chord functions. Processing of high and low cloze probability words as indexed by the N400 was not affected by the presentation of irregular chord functions. In a control experiment, the LAN was not affected by physically deviant tones that elicited a mismatch negativity (MMN). Results demonstrate that processing of musical syntax (as reflected in the ERAN) interacts with the processing of linguistic syntax (as reflected in the LAN), and that this interaction is not due to a general effect of deviance-related negativities that precede an LAN. Findings thus indicate a strong overlap of neural resources involved in the processing of syntax in language and music. PMID- 16269098 TI - Automatic encoding of polyphonic melodies in musicians and nonmusicians. AB - In music, multiple musical objects often overlap in time. Western polyphonic music contains multiple simultaneous melodic lines (referred to as "voices") of equal importance. Previous electrophysiological studies have shown that pitch changes in a single melody are automatically encoded in memory traces, as indexed by mismatch negativity (MMN) and its magnetic counterpart (MMNm), and that this encoding process is enhanced by musical experience. In the present study, we examined whether two simultaneous melodies in polyphonic music are represented as separate entities in the auditory memory trace. Musicians and untrained controls were tested in both magnetoencephalogram and behavioral sessions. Polyphonic stimuli were created by combining two melodies (A and B), each consisting of the same five notes but in a different order. Melody A was in the high voice and Melody B in the low voice in one condition, and this was reversed in the other condition. On 50% of trials, a deviant final (5th) note was played either in the high or in the low voice, and it either went outside the key of the melody or remained within the key. These four deviations occurred with equal probability of 12.5% each. Clear MMNm was obtained for most changes in both groups, despite the 50% deviance level, with a larger amplitude in musicians than in controls. The response pattern was consistent across groups, with larger MMNm for deviants in the high voice than in the low voice, and larger MMNm for in-key than out-of-key changes, despite better behavioral performance for out-of-key changes. The results suggest that melodic information in each voice in polyphonic music is encoded in the sensory memory trace, that the higher voice is more salient than the lower, and that tonality may be processed primarily at cognitive stages subsequent to MMN generation. PMID- 16269099 TI - Who's in control? Proficiency and L1 influence on L2 processing. AB - We report three reaction time (RT)/event-related brain potential (ERP) semantic priming lexical decision experiments that explore the following in relation to L1 activation during L2 processing: (1) the role of L2 proficiency, (2) the role of sentence context, and (3) the locus of L1 activations (orthographic vs. semantic). All experiments used German (L1) homonyms translated into English (L2) to form prime-target pairs (pine-jaw for Kiefer) to test whether the L1 caused interference in an all-L2 experiment. Both RTs and ERPs were measured on targets. Experiment 1 revealed reversed priming in the N200 component and RTs for low proficiency learners, but only RT interference for high-proficiency participants. Experiment 2 showed that once the words were processed in sentence context, the low-proficiency participants still showed reversed N200 and RT priming, whereas the high-proficiency group showed no effects. Experiment 3 tested native English speakers with the words in sentence context and showed a null result comparable to the high-proficiency group. Based on these results, we argue that cognitive control relating to translational activation is modulated by (1) L2 proficiency, as the early interference in the N200 was observed only for low-proficiency learners, and (2) sentence context, as it helps high-proficiency learners control L1 activation. As reversed priming was observed in the N200 and not the N400 component, we argue that (3) the locus of the L1 activations was orthographic. Implications in terms of bilingual word recognition and the functional role of the N200 ERP component are discussed. PMID- 16269100 TI - Specificity of the effect of a nicotinic receptor polymorphism on individual differences in visuospatial attention. AB - Cortical neurotransmitter availability is known to exert domain-specific effects on cognitive performance. Hence, normal variation in genes with a role in neurotransmission may also have specific effects on cognition. We tested this hypothesis by examining associations between polymorphisms in genes affecting cholinergic and noradrenergic neurotransmission and individual differences in visuospatial attention. Healthy individuals were administered a cued visual search task which varied the size of precues to the location of a target letter embedded in a 15-letter array. Cues encompassed 1, 3, 9, or 15 letters. Search speed increased linearly with precue size, indicative of a spatial attentional scaling mechanism. The strength of attentional scaling increased progressively with the number of C alleles (0, 1, or 2) of the alpha-4 nicotinic receptor gene C1545T polymorphism (n = 104). No association was found for the dopamine beta hydroxylase gene G444A polymorphism (n = 135). These findings point to the specificity of genetic neuromodulation. Whereas variation in a gene linked to cholinergic transmission systematically modulated the ability to scale the focus of visuospatial attention, variation in a gene governing dopamine availability did not. The results show that normal variation in a gene controlling a nicotinic receptor makes a selective contribution to individual differences in visuospatial attention. PMID- 16269102 TI - Effects of vestibular rotatory accelerations on covert attentional orienting in vision and touch. AB - Peripheral vestibular organs feed the central nervous system with inputs favoring the correct perception of space during head and body motion. Applying temporal order judgments (TOJs) to pairs of simultaneous or asynchronous stimuli presented in the left and right egocentric space, we evaluated the influence of leftward and rightward vestibular rotatory accelerations given around the vertical head body axis on covert attentional orienting. In a first experiment, we presented visual stimuli in the left and right hemifield. In a second experiment, tactile stimuli were presented to hands lying on their anatomical side or in a crossed position across the sagittal body midline. In both experiments, stimuli were presented while normal subjects suppressed or did not suppress the vestibulo ocular response (VOR) evoked by head-body rotation. Independently of VOR suppression, visual and tactile stimuli presented on the side of rotation were judged to precede simultaneous stimuli presented on the side opposite the rotation. When limbs were crossed, attentional facilitatory effects were only observed for stimuli presented to the right hand lying in the left hemispace during leftward rotatory trials with VOR suppression. This result points to spatiotopic rather than somatotopic influences of vestibular inputs, suggesting that cross-modal effects of these inputs on tactile ones operate on a representation of space that is updated following arm crossing. In a third control experiment, we demonstrated that temporal prioritization of stimuli presented on the side of rotation was not determined by response bias linked to spatial compatibility between the directions of rotation and the directional labels used in TOJs (i.e., "left" or "right" first). These findings suggest that during passive rotatory head-body accelerations, covert attention is shifted toward the direction of rotation and the direction of the fast phases of the VOR. PMID- 16269101 TI - Comparing the effects of auditory deprivation and sign language within the auditory and visual cortex. AB - To investigate neural plasticity resulting from early auditory deprivation and use of American Sign Language, we measured responses to visual stimuli in deaf signers, hearing signers, and hearing nonsigners using functional magnetic resonance imaging. We examined "compensatory hypertrophy" (changes in the responsivity/size of visual cortical areas) and "cross-modal plasticity" (changes in auditory cortex responses to visual stimuli). We measured the volume of early visual areas (V1, V2, V3, V4, and MT+). We also measured the amplitude of responses within these areas, and within the auditory cortex, to a peripheral visual motion stimulus that was attended or ignored. We found no major differences between deaf and hearing subjects in the size or responsivity of early visual areas. In contrast, within the auditory cortex, motion stimuli evoked significant responses in deaf subjects, but not in hearing subjects, in a region of the right auditory cortex corresponding to Brodmann's areas 41, 42, and 22. This hemispheric selectivity may be due to a predisposition for the right auditory cortex to process motion; earlier studies report a right hemisphere bias for auditory motion in hearing subjects. Visual responses within the auditory cortex of deaf subjects were stronger for attended than ignored stimuli, suggesting top-down processes. Hearing signers did not show visual responses in the auditory cortex, indicating that cross-modal plasticity can be attributed to auditory deprivation rather than sign language experience. The largest effects of auditory deprivation occurred within the auditory cortex rather than the visual cortex, suggesting that the absence of normal input is necessary for large-scale cortical reorganization to occur. PMID- 16269103 TI - Does prosopagnosia take the eyes out of face representations? Evidence for a defect in representing diagnostic facial information following brain damage. AB - One of the most impressive disorders following brain damage to the ventral occipitotemporal cortex is prosopagnosia, or the inability to recognize faces. Although acquired prosopagnosia with preserved general visual and memory functions is rare, several cases have been described in the neuropsychological literature and studied at the functional and neural level over the last decades. Here we tested a brain-damaged patient (PS) presenting a deficit restricted to the category of faces to clarify the nature of the missing and preserved components of the face processing system when it is selectively damaged. Following learning to identify 10 neutral and happy faces through extensive training, we investigated patient PS's recognition of faces using Bubbles, a response classification technique that sampled facial information across the faces in different bandwidths of spatial frequencies [Gosselin, F., & Schyns, P. E., Bubbles: A technique to reveal the use of information in recognition tasks. Vision Research, 41, 2261-2271, 2001]. Although PS gradually used less information (i.e., the number of bubbles) to identify faces over testing, the total information required was much larger than for normal controls and decreased less steeply with practice. Most importantly, the facial information used to identify individual faces differed between PS and controls. Specifically, in marked contrast to controls, PS did not use the optimal eye information to identify familiar faces, but instead the lower part of the face, including the mouth and the external contours, as normal observers typically do when processing unfamiliar faces. Together, the findings reported here suggest that damage to the face processing system is characterized by an inability to use the information that is optimal to judge identity, focusing instead on suboptimal information. PMID- 16269104 TI - Neural correlates of syntactic processing in two-year-olds. AB - Event-related brain potential (ERP) studies of sentence processing in adults have shown that phrase-structure violations are associated with two ERP components: an early left anterior negativity (ELAN) and a late, centro-parietal positivity (P600). Although the ELAN reflects highly automatic first-pass sentence parsing, the P600 has been interpreted to reflect later, more controlled processes. The present ERP study investigates the processing of phrase-structure violations in children below three years of age. Both children (mean age of 2.8 years) and adults passively listened to short active sentences that were either correct or syntactically incorrect. Adults displayed an ELAN that was followed by a P600 to the syntactic violation. Children also demonstrated a biphasic ERP pattern consisting of an early left hemispheric negativity and a late positivity. Both components, however, started later and persisted longer than those observed in adults. The left lateralization of the children's negativity suggests that this component can be interpreted as a child-specific precursor to the ELAN observed in adults. The appearance of the early negativity indicates that the neural mechanisms of syntactic parsing are present, in principle, during early language development. PMID- 16269105 TI - Delay-period activity in the prefrontal cortex: one function is sensory gating. AB - The prefrontal cortex (PFC) contributes to working memory functions via executive control processes that do not entail the storage, per se, of mnemonic representations. One of these control processes may be a sensory gating mechanism that facilitates retention of representations in working memory by down regulating the gain of the sensory processing of intervening irrelevant stimuli. This idea was tested by scanning healthy young adults with functional magnetic resonance imaging while they performed a delayed face-recognition task. The 2 x 2 factorial design varied the factors of Memory (present, absent) and Distraction (present, absent). During memory-present trials, target and probe stimuli were individual gray-scale male faces. Memory-absent trials were identical, except that they employed the same recurring female faces (denoting a "no memory" trial). Distraction-present trials featured rapid serial visual presentation of bespectacled male faces during the two middle seconds of the delay. The first step of the analyses identified dorsolateral PFC (dlPFC) and inferior occipitotemporal cortex (IOTC) voxels exhibiting delay-period activity in memory present/distraction-absent trials, that is, the "unfilled" delay. Within these voxels, distraction-evoked activity in the dlPFC was markedly higher during trials that required the concurrent short-term retention of information than on those that did not, whereas the opposite effect was seen in the IOTC. These results are consistent with the view that processes related to sensory gating account for a portion of the delay-period activity that is routinely observed in the dlPFC. PMID- 16269107 TI - Familiarity affects the processing of task-irrelevant auditory deviance. AB - The effects of familiarity on auditory change detection on the basis of auditory sensory memory representations were investigated by presenting oddball sequences of sounds while participants ignored the auditory stimuli. Stimulus sequences were composed of sounds that were familiar and sounds that were made unfamiliar by playing the same sounds backward. The roles of frequently presented stimuli (standards) and infrequently presented ones (deviants) were fully crossed. Deviants elicited the mismatch negativity component of the event-related brain potential. We found an enhancement in detecting changes when deviant sounds appeared among familiar standard sounds compared when they were delivered among unfamiliar standards. Familiarity with the deviant sounds also enhanced the change-detection process. We suggest that tuning to familiar items sets up preparatory processes that affect change detection in familiar sound sequences. PMID- 16269108 TI - Correlates of capture of attention and inhibition of return across stages of visual processing. AB - How do visual signals evolve from early to late stages in sensory processing? We explored this question by examining two neural correlates of spatial attention. The capture of attention and inhibition of return refer to the initial advantage and subsequent disadvantage to respond to a visual target that follows an irrelevant visual cue at the same location. In the intermediate layers of the superior colliculus (a region that receives input from late stages in visual processing), both behavioral effects link to changes in the neural representation of the target: strong target-related activity correlates with the capture of attention and weak target-related activity correlates with inhibition of return. Contrasting these correlates with those obtained in the superficial layers (a functionally distinct region that receives input from early stages in visual processing), we show that the target-related activity of neurons in the intermediate layers was the best predictor of orienting behavior, although dramatic changes in the target-related response were observed in both subregions. We describe the important consequences of these findings for understanding the neural basis of the capture of attention and inhibition of return and interpreting changes in neural activity more generally. PMID- 16269106 TI - Dynamics of a temporo-fronto-parietal network during sustained spatial or spectral auditory processing. AB - Animal and human studies have suggested that posterior temporal, parietal, and frontal regions are specifically involved in auditory spatial (location and motion) processing, forming a putative dorsal "where" pathway. We used scalp EEG and current density mapping to investigate the dynamics of this network in human subjects presented with a varying acoustic stream in a two-factor paradigm: spatial versus pitch variations, focused versus diverted attention. The main findings were: (i) a temporo-parieto-frontal network was activated during the whole duration of the stream in all conditions and modulated by attention; (ii) the left superior temporal cortex was the only region showing different activations for pitch and spatial variations. Therefore, parietal and frontal regions would be involved in task-related processes (attention and motor preparation), whereas the differential processing of acoustic spatial and object related features seems to take place at the temporal level. PMID- 16269109 TI - FROST: a distributed neurocomputational model of working memory maintenance. AB - Many studies suggest that the sustained activation underlying working memory (WM) maintenance is mediated by a distributed network that includes the prefrontal cortex and other structures (e.g., posterior parietal cortex, thalamus, globus pallidus, and the caudate nucleus). A computational model of WM, called FROST (short for FROntal-Striatal-Thalamic), is proposed in which the representation of items and spatial positions is encoded in the lateral prefrontal cortex. During delay intervals, activation in these prefrontal cells is sustained via parallel, prefrontal cortical-thalamic loops. Activation reverberates in these loops because prefrontal cortical excitation of the head of the caudate nucleus leads to disinhibition of the thalamus (via the globus pallidus). FROST successfully accounts for a wide variety of WM data, including single-cell recording data and human behavioral data. PMID- 16269110 TI - Regional brain activation evoked when approaching a virtual human on a virtual walk. AB - We investigated the necessity of biological motion for activation of the posterior superior temporal sulcus (pSTS) in circumstances in which the rapid approach of the observer to a virtual human induced the observer to make inferences about the characters intentions. Using a virtual reality environment, subjects experienced themselves walking towards a complex scene composed of animate and/or inanimate objects. During "person" trials, the scene contained a virtual human either making a simple gesture such as scratching his face (Study 1) or standing completely still (Study 2). During "object" trials, the scenes contained items such as furniture, a face portrait, and a clock, but not the virtual human. Using functional MRI to measure brain activity, we demonstrated strong activity in the pSTS while the observer approached the social scene, but only when the virtual human was making gestures. This result emphasizes the importance of biological motion in inferring the intentions of others. PMID- 16269111 TI - Dissociating reading processes on the basis of neuronal interactions. AB - Previous studies of patients with phonological and surface alexia have demonstrated a double dissociation between the reading of pseudo words and words with atypical spelling-to-sound relationships. A corresponding double dissociation in the neuronal activation patterns for pseudo words and exception words has not, however, been consistently demonstrated in normal subjects. Motivated by the literature on acquired alexia, the present study contrasted pseudo words to exception words and explored how neuronal interactions within the reading system are influenced by word type. Functional magnetic resonance imaging was used to measure neuronal responses during reading in 22 healthy volunteers. The direct comparison of reading pseudo words and exception words revealed a double dissociation within the left frontal cortex. Pseudo words preferentially increased left dorsal premotor activation, whereas exception words preferentially increased left pars triangularis activation. Critically, these areas correspond to those previously associated with phonological and semantic processing, respectively. Word-type dependent interactions between brain areas were then investigated using dynamic causal modeling. This revealed that increased activation in the dorsal premotor cortex for pseudo words was associated with a selective increase in effective connectivity from the posterior fusiform gyrus. In contrast, increased activation in the pars triangularis for exception words was associated with a selective increase in effective connectivity from the anterior fusiform gyrus. The present investigation is the first to identify distinct neuronal mechanisms for semantic and phonological contributions to reading. PMID- 16269112 TI - When blue is larger than red: colors influence numerical cognition in synesthesia. AB - In synesthesia, certain stimuli ("inducers") may give rise to perceptual experience in additional modalities not normally associated with them ("concurrent"). For example, color-grapheme synesthetes automatically perceive achromatic numbers as colored (e.g., 7 is turquoise). Although synesthetes know when a given color matches the one evoked by a certain number, colors do not automatically give rise to any sort of number experience. The behavioral consequences of synesthesia have been documented using Stroop-like paradigms, usually using color judgments. Owing to the unidirectional nature of the synesthetic experience, little has been done to obtain performance measures that could indicate whether bidirectional cross-activation occurs in synesthesia. Here it is shown that colors do implicitly evoke numerical magnitudes in color grapheme synesthetes, but not in nonsynesthetic participants. It is proposed that bidirectional coactivation of brain areas is responsible for the links between color and magnitude processing in color-grapheme synesthesia and that unidirectional models of synesthesia might have to be revised. PMID- 16269113 TI - Where memory meets attention: neural substrates of negative priming. AB - The negative priming (NP) effect refers to the observed increase in identification time for a current target stimulus or stimulus feature (the "probe") that has been employed as a distractor stimulus or stimulus feature on the previous trial (the "prime"), representing strong evidence that ignored information is actively processed to a high level by selective attention systems. However, theoretical accounts of NP differ in whether they attribute the effect to processes of selective inhibition or episodic memory retrieval. Here we derived neurophysiological predictions from the rival "selective inhibition" and "episodic retrieval" models of NP, and employed event-related fMRI in a color naming Stroop task to assess neural responses to probe trials that were subject to either no priming or negative priming. Compared to no-priming probe trials, NP resulted in increased activation of the right dorsolateral prefrontal cortex, in a region which has been closely linked with episodic memory retrieval functions. NP was also accompanied by activation of the right thalamus, particularly the mediodorsal nucleus, which has been implicated in the pathophysiology of schizophrenia, a condition associated with diminished NP effects. Our results support the proposal that ignored stimulus information is fully encoded in memory, and that episodic retrieval, not selective inhibition, of such information affects selective attention performance on subsequent trials. PMID- 16269114 TI - Phonotactic knowledge and lexical-semantic processing in one-year-olds: brain responses to words and nonsense words in picture contexts. AB - During their first year of life, infants not only acquire probabilistic knowledge about the phonetic, prosodic, and phonotactic organization of their native language, but also begin to establish first lexical-semantic representations. The present study investigated the sensitivity to phonotactic regularities and its impact on semantic processing in 1-year-olds. We applied the method of event related brain potentials to 12- and 19-month-old children and to an adult control group. While looking at pictures of known objects, subjects listened to spoken nonsense words that were phonotactically legal (pseudowords) or had phonotactically illegal word onsets (nonwords), or to real words that were either congruous or incongruous to the picture contents. In 19-month-olds and in adults, incongruous words and pseudowords, but not nonwords, elicited an N400 known to ref lect mechanisms of semantic integration. For congruous words, the N400 was attenuated by semantic priming. In contrast, 12-month-olds did not show an N400 difference, neither between pseudo- and nonwords nor between incongruous and congruous words. Both 1-year-old groups and adults additionally displayed a lexical priming effect for congruous words, that is, a negativity starting around 100 msec after words onset. One-year-olds, moreover, displayed a phonotactic familiarity effect, that is, a widely distributed negativity starting around 250 msec in 19-month-olds but occurring later in 12-month-olds. The results imply that both lexical priming and phonotactic familiarity already affect the processing of acoustic stimuli in children at 12 months of age. In 19-month-olds, adult-like mechanisms of semantic integration are present in response to phonotactically legal, but not to phonotactically illegal, nonsense words, indicating that children at this age treat pseudowords, but not nonwords, as potential word candidates. PMID- 16269115 TI - Early event-related potential effects of syllabic processing during visual word recognition. AB - A number of behavioral studies have suggested that syllables might play an important role in visual word recognition in some languages. We report two event related potential (ERP) experiments using a new paradigm showing that syllabic units modulate early ERP components. In Experiment 1, words and pseudowords were presented visually and colored so that there was a match or a mismatch between the syllable boundaries and the color boundaries. The results showed color syllable congruency effects in the time window of the P200. Lexicality modulated the N400 amplitude, but no effects of this variable were obtained at the P200 window. In Experiment 2, high- and low-frequency words and pseudowords were presented in the congruent and incongruent conditions. The results again showed congruency effects at the P200 for low-frequency words and pseudowords, but not for high-frequency words. Lexicality and lexical frequency effects showed up at the N400 component. The results suggest a dissociation between syllabic and lexical effects with important consequences for models of visual word recognition. PMID- 16269116 TI - Salvadoran fathers' attendance at prenatal care, delivery, and postpartum care. AB - OBJECTIVE: To provide a baseline perspective on the prevalence of Salvadoran men's attendance at prenatal care, delivery, and postpartum well-baby care and on sociodemographic factors associated with their attendance, with the goal of informing efforts to help men play more positive roles in maternal-child health. METHODS: The data came from the 2003 Salvadoran National Male Health Survey. The data focused on fathers (n = 418) and their most recent live-born child in the preceding five years. Factors associated with the fathers' participation in prenatal care visits, attendance at delivery, and participation in postnatal well baby visits were explored using logistic and multinomial regression models. RESULTS: Ninety percent of the recent Salvadoran fathers who were surveyed participated in a prenatal care visit, attended the delivery, or participated in a postpartum well-baby care visit; 34% participated in all three of the activities. Attendance at delivery was most common, reported by 81% of fathers; the most common reason that subjects cited for not attending was that they had had to work. CONCLUSIONS: A large majority of the Salvadoran fathers participated in at least one prenatal care visit, delivery, or a postpartum well-baby care visit. While attendance alone does not necessarily indicate that men are supporting their partners, the results suggest that norms are in place for men to play positive roles in maternal-child health matters. Furthermore, the participation of fathers in these maternal and child health care activities may provide new opportunities to educate and further support men in both their own health and their family's health. PMID- 16269117 TI - [Differences in the prevalence of malnutrition resulting from using a national growth reference or an international growth reference]. AB - OBJECTIVE: To evaluate if using an international growth reference versus a national growth reference leads to differences in the prevalences of low weight for-height (wasting) and low height-for-age (stunting) in a sample of children 8 to 14 years old. METHODS: A cross-sectional study was carried out with a sample of 1,470 school-children (both boys and girls) in the city of La Plata, in the province of Buenos Aires, Argentina. Based on the data on weight and height, Z scores were calculated with regard to the reference values from the United States of America's first and second National Health and Nutrition Examination Survey (NHANES I and NHANES II), and the reference values for Argentina from the Argentine Pediatric Society. The prevalences of wasting and stunting were calculated, using as the cutoff point two standard deviations below the median of the reference. The prevalences were compared using the chi-squared test with Yates' correction, and odds ratios (ORs) and 95% confidence intervals (95% CIs) were calculated. RESULTS: The international reference was positively associated with a higher prevalence of stunting in the entire sample (OR = 5.12; 95% CI: 3.27 to 8.05), in the boys (OR = 4.36; 95% CI: 2.43 to 7.93), and in the girls (OR = 6.32; 95% CI: 3.10 to 13.27). The prevalence of wasting was similar with both references. The distribution of height-for-age was skewed to the left with respect to the international reference and to the right with respect to the Argentine reference, while the distribution of weight-for-height overlapped both references. CONCLUSIONS: The differences in the prevalences of stunting calculated according to the two references that were used indicate a different nutritional status for the population studied. The increase in the height of the children with respect to the Argentine reference points to the presence of a positive secular trend, and the increase can explain part of the differences in the prevalence of stunting. PMID- 16269118 TI - A literature review of the epidemiology of sepsis in Latin America. AB - OBJECTIVES: Epidemiological studies from the United States of America and from Europe have shown that sepsis is a widely prevalent syndrome, with either steady or slightly decreasing rates of morbidity and of mortality in recent decades. The objective of this paper is to provide a systematic review regarding the description and characterization of sepsis in Latin America. METHODS: To locate materials on sepsis in Latin America, a comprehensive search strategy was employed with three medical bibliographic databases, using combinations of the terms "sepsis," "septicemia," "bacteremia," "sepsis syndrome," "epidemiology," "incidence," and "prevalence." The materials selected were in English, Spanish, or Portuguese. RESULTS: The titles of more than 1,000 potentially relevant articles were screened, and more than 600 selected abstracts were reviewed in detail. Twenty papers published from 1990 through 2004 were selected and analyzed. The studies described in the 20 articles were extremely heterogeneous in design, population, sample size, end points, and follow-up. The studies did not all apply the same clinical definition for sepsis, thus making it impossible to produce a precise overall estimate of the magnitude of the problem of sepsis in Latin America. CONCLUSIONS: The results of the literature review suggest that the clinical and epidemiological approaches to the problem of sepsis in Latin America have sometimes been inappropriate with respect to research design, study population, and clinical outcome. Further, some data suggest that in terms of both frequency and mortality the situation with sepsis and severe systemic infections may be worse in Latin America than it is in developed countries. PMID- 16269119 TI - [Usage of food and beverage labels by supermarket shoppers in Brasilia, Brazil]. AB - OBJECTIVE: To investigate whether adults who shop in supermarkets in one part of Brasilia, the capital city of Brazil, use the information contained in food and beverage labels, as well as to characterize this usage. METHODS: This study was done in five supermarkets in the Plano Piloto (Pilot Plan) central core area of Brasilia. The research was carried out in two stages: (1) a quantitative stage, based on a cross-sectional study, during which 250 individuals were randomly selected and surveyed in the supermarkets; and (2) a qualitative stage, in which 25 individuals who had participated in the first stage underwent more extensive individual interviews. RESULTS: Of the 250 persons surveyed, 187 of them (74.8%) reported that they usually read food and beverage labels. However, only 25.7% of the consumers who consulted the labels did so for all foods and beverages. Of the persons who read the labels, the majority (59.9%) only read the labels for specific foods, including milk, dairy products, canned foods, sausages, and diet products. The information that shoppers most often wanted to know from the labels was the number of calories and the fat and sodium content. CONCLUSIONS: Our study shows the need to utilize our findings in improving the existing educational strategies for promoting healthy eating. We suggest that consumers, food producers, and food distributors all be involved in developing strategies for nutrition education. PMID- 16269120 TI - Assessing the causes of under-five mortality in the Albert Schweitzer Hospital service area of rural Haiti. AB - OBJECTIVES: Limited information is available regarding the causes of under-five mortality in nearly all of the countries in which mortality is the highest. The purpose of this study was to use a standard computerized protocol for defining the leading causes of death among children in a high-mortality rural population of Haiti and to highlight the need for similar studies else-where in Haiti and throughout the high-mortality areas of Latin America and the Caribbean. METHODS: In 2001 a standardized, closed-ended verbal autopsy questionnaire endorsed by the World Health Organization was administered to a representative, population-based sample of the mothers or other caregivers of 97 children who had died before reaching 5 years of age between 1995 and 1999 in the service area of the Albert Schweitzer Hospital, which is located in the rural Artibonite Valley of Haiti. With the data from the questionnaires we used a computerized algorithm to generate diagnoses of the cause of death; the algorithm made it possible to have more than one cause of death. RESULTS: Acute lower respiratory infection (ALRI) was the leading diagnosis, present in 45% of all under-five deaths, followed by enteric diseases, present in 21% of deaths. Neonatal tetanus, preterm birth, and other early neonatal causes unassociated with ALRI or diarrhea were present in 41% of the neonatal deaths. Among children 1-59 months of age, ALRI was present in 51% of the deaths, and enteric diseases in 30%. Deaths were concentrated during the first few months of life, with 35% occurring during the first month. Among the neonatal deaths, 27% occurred on the first day of life, and 80% occurred during the first 10 days of life. CONCLUSIONS: In the Albert Schweitzer Hospital program area--and presumably in other areas of Haiti as well--priority needs to be given to the prevention of and the early, effective treatment of ALRI, diarrhea, and early neonatal conditions. This study points to the need for more, similar standardized assessments to guide local, regional, and national programs. PMID- 16269121 TI - [Determining exposure history in occupational epidemiology]. AB - In epidemiology, it is necessary that exposure indicators have good validity in order to obtain valid results when measuring the risks associated with occupational exposure to environmental noxious agents. However, ensuring the validity of past exposure data is no easy task. Because there are no environmental hygiene measures or representative levels of bioindicators signaling past exposure, self-reports have been used as a source of indirect exposure data. Unfortunately, data on specific agents are commonly poor and need to be complemented with data on the determinants of exposure. The validity of self-reports improves when certain techniques, such as control lists and icons, are employed, and the quality of individual exposure data improves when secondary data on exposure and its conditioning or determining factors are incorporated. Exposure can be determined by means of exposure matrices, assessment by experts, and exposure models, and by using a combination of primary and secondary data on exposure and its conditioning factors. Matrices contain pooled data and can thus lead to errors in classifying individual exposure and to biased risk estimates. Assessment by experts is probably the method with the highest validity, but it can become expensive when studies are large. It is also feasible to use a formal model for assessing perceivable exposures, complemented with expert assessments whenever the results of the model appear to deviate from reality. PMID- 16269122 TI - Global child survival initiatives and their relevance to the Latin American and Caribbean Region. AB - We review two series of papers published by The Lancet: the Child Survival Series (2003) and the Neonatal Survival Series (2005). Both series drew attention to the nearly 11 million annual deaths of children under the age of five years, and to the fact that almost 4 million of these deaths occur in the first month of life. We show that two thirds of these deaths could be prevented through universal coverage with existing, low-cost interventions that are failing to reach most children in the world. The series also highlighted the importance of reducing inequities both between and within countries. The relevance of these series to Latin America and the Caribbean is examined. Although substantial progress has been made in reducing mortality and improving coverage, two major challenges remain: how to improve the quality of health interventions, and how to reach the most disadvantaged children in the Latin American and Caribbean Region. PMID- 16269123 TI - [Preventing influenza: recommendations for the 2005-2006 season]. PMID- 16269124 TI - Public health impact and medical consequences of earthquakes. PMID- 16269125 TI - A genetically encoded indicator for assaying bioactive chemicals that induce nuclear transport of glucocorticoid receptor. AB - Glucocorticoids, the adrenal steroid hormones secreted during stress, are essential to homeostasis and metabolism in the human body. An impaired glucocorticoid signaling due to dysfunction of the glucocorticoid receptor (GR) by synthetic chemicals can cause diseases and disruptions of the homeostasis and metabolism. Here we demonstrate the development of a method for screening endocrine-disrupting chemicals and potent risk factors of human diseases based on the nuclear trafficking of the GR. We constructed a new assay using a pair of genetic indicators with the full length of the GR, split Renilla luciferase (RLuc), and split DnaE (a protein splicing element). The GR-containing fusion protein with C-terminal halves of DnaE and RLuc is localized in cytosol due to the cytosolic character of the GR, whereas the fusion protein with N-terminal halves of DnaE and RLuc stays in the nucleus due to the cofused nucleus localization signal. On being stimulated with a ligand, the GR is translocated into the cellular nucleus. Thus, a protein splicing occurs in the nucleus by an interaction between the splicing junctions of each DnaE fragment. The enzymatic activities from the reconstituted RLuc allow the ligand-dependent luminescence intensities. The feasibility of the method was evaluated by quantifying the hormonal activities of 20 different kinds of steroids and synthetic chemicals using the NIH 3T3 cells carrying the pair of indicators. The hormonal activities of tested ligands are discussed based on the chemical structure-activity relationship. We found that androgens, testosterone, and 19-nortestosterone weakly induce the nuclear transport of the GR. The current assay allows high throughput screening of risk chemicals and drug candidates influential to a signal transduction pathway of the GR. PMID- 16269126 TI - Beta-D-glucosidase reaction kinetics from isothermal titration microcalorimetry. AB - The cellobiase activities of nine thermal stable mutants of Thermobifida fusca BglC were assayed by isothermal titration microcalorimetry (ITC). The mutations were previously generated using random mutagenesis and identified by high temperature screening as imparting improved thermal stability to the beta-D glucosidase enzyme. Analysis of the substrate-saturation curves obtained by ITC for the wild-type enzyme and the nine thermally stabilized mutants revealed that the wild type and all the mutants were subject to binding of a second substrate molecule. Furthermore, the "inhibited" enzyme-substrate complexes were shown to retain catalytic activity. In the case of three of the BglC mutants (N178I, N317Y/L444F, and N317Y/L444F/A433V), binding of a second substrate molecule resulted in improved cellobiose turnover rates at lower substrate concentrations. No correlation between denaturation temperatures of the mutants and activity on cellobiose at 25 degrees C was evident. However, one particular mutant, BglC S319C, was significantly improved in both thermal tolerance and cellobiase activity with respect to those of the wild-type BglC. The triple mutant, N317Y/L444F/A433V, had a 5 degrees C increase in denaturation temperature while maintaining activity levels similar to that of the wild type at higher substrate concentrations. ITC provided a highly sensitive and nondestructive means to continuously monitor the reaction of BglC with cellobiose, resulting in abundant data sets that could be rigorously analyzed by fitting to known enzyme kinetics models. One distinct advantage of using data from the ITC was the empirical validation of the pseudo steady state assumption, a necessary condition for obtaining solutions to the proposed mechanisms. PMID- 16269127 TI - Mass spectrometric identification and quantification of glycosyl flavonoids, including dihydrochalcones with neutral loss scan mode. AB - We developed a strategy for determination and quantification of glycosyl flavonoids using liquid chromatography-triple quadrupole mass spectrometry with neutral loss scan at 15 and 30eV collision energy in the positive ion mode. The fragmentation patterns of glycosyl flavonoids at 15 and 30eV showed that fragmentation of sugar moiety depended on the type of glycosidic bond to aglycone, the site of C-glycosylation, and the type of aglycone. C-Glycosyl dihydrochalcones especially stood out because they produced [M+H-162](+) even at 15eV such as O-glycoside in spite of C-glycoside. C-Glycosides were classified according to (i) the intensity ratio A of [M+H-150](+) to [M+H-120](+) at 30eV and (ii) the intensity ratio B of [M+H-120](+) at 15eV to one at 30eV. The 8-C glycosides were A<1 and B<1, the 6-C-glycosides were A>1 and B<1, and the C glycosyl dihydrochalcones were A>1 and B>>1. Therefore, the intensity ratios of the neutral loss scan of 120 and 150Da at 30eV and those of 120, 162, and 308Da at 15eV allowed sequential distinction among these three types of C-glycosides as well as between O- and C-glycosides. Our method was applied for analysis of Rooibos tea, and the identified glycosides could be quantified specifically by the selected reaction monitoring method. PMID- 16269128 TI - Distribution of 10 periodontal bacteria in saliva samples from Japanese children and their mothers. AB - OBJECTIVE: We analyzed the distribution of 10 periodontal bacteria species (Porphyromonas gingivalis, Tannerella forsythensis, Prevotella intermedia, Prevotella nigrescens, Campylobacter rectus, Eikenella corrodens, Actinobacillus actinomycetemcomitans, Capnocytophaga ochracea, Capnocytophaga sputigena, and Treponema denticola) in children, and then compared their distribution in those children and their mothers, with special attention given to three of the species known as the red complex (P. gingivalis, T. forsythensis, and T. denticola) whose presence has been shown to be associated with conditions related to periodontal diseases. METHODS: One hundred thirteen pairs of children and their mothers were randomly selected from patients treated at the Pedodontic Clinic of Osaka University Dental Hospital. Saliva samples were taken at the second visit prior to receiving professional tooth brushing instruction. Genomic DNA was extracted from each saliva sample, followed by a polymerase chain reaction assay with species-specific sets of primers. RESULTS: A. actinomycetemcomitans was the most frequently detected species in the mothers, followed by C. sputigena, P. gingivalis, and T. forsythensis, while C. sputigena had the highest detection rate, followed by A. actinomycetemcomitans and T. denticola in the children. The detection rate of the red complex species in children whose mothers possessed the same species was significantly higher than in those whose mothers did not possess them. CONCLUSIONS: Our results indicate a correlation between the presence of periodontal bacteria in children and their mothers, while the presence of red complex bacteria in children was highly associated with that in their mothers. PMID- 16269129 TI - Endothelial stress induces the release of vitamin D-binding protein, a novel growth factor. AB - Endothelial cells (EC) under stress release paracrine mediators that facilitate accumulation of vascular smooth muscle cells (VSCM) at sites of vascular injury. We found that medium conditioned by serum-starved EC increase proliferation and migration of VSCM in vitro. Fractionation of the conditioned medium followed by mass spectral analysis identified one bioactive component as vitamin D-binding protein (DBP). DBP induced both proliferation and migration of VSMC in vitro in association with increased phosphorylation of ERK 1/2. PD 98059, a biochemical inhibitor of ERK 1/2, abrogated these proliferative and migratory responses in VSMC. DBP is an important carrier for the vitamin-D sterols, 25-hydroxyvitamin-D, and 1alpha,25-dihydroxyvitamin-D. Both sterols inhibited the activity of DBP on VSMC, suggesting that vitamin D binding sites are important for initiating the activities of DBP on VSMC. Release of DBP at sites of endothelial injury represents a novel pathway favoring accumulation of VSMC at sites of vascular injury. PMID- 16269130 TI - Exposure of protein kinase motifs that trigger binding of Hsp90 and Cdc37. AB - Hsp90 and its co-chaperone Cdc37 are required for the activity of numerous eukaryotic protein kinases. c-Jun N-terminal kinases (JNKs) appear to be Hsp90 independent kinases, as their activity is unaffected by Hsp90 inhibition. It is currently unknown why some protein kinases are Hsp90- and Cdc37-dependent for their function, while others are not. Therefore, we investigated what structural motifs within JNKs confer or defer Hsp90 and Cdc37 interaction. Both Hsp90 and Cdc37 recognized structural features that were exposed or destabilized upon deletion of JNK1alpha1's N-terminal non-catalytic structural motif, while only Hsp90 bound JNK when its C-terminal non-catalytic structural motif was deleted. Mutations in JNK's activation loop that are known to constitutively activate or inactivate its kinase activity had no effect on JNK's lack of interaction with Hsp90 and Cdc37. Our findings suggest a model in which Hsp90 and Cdc37 each recognize distinct features within the catalytic domains of kinases. PMID- 16269131 TI - Structure of Thermus thermophilus type 2 isopentenyl diphosphate isomerase inferred from crystallography and molecular dynamics. AB - Crystal structures of Thermus thermophilus and Bacillus subtilis type 2 IPP isomerases were combined to generate an almost complete model of the FMN-bound structure of the enzyme. In contrast to previous studies, positions of flexible loops were obtained and carefully analyzed by molecular dynamics. Docking simulations find a unique putative binding site for the IPP substrate. PMID- 16269132 TI - Polyamine sensing during antizyme mRNA programmed frameshifting. AB - A key regulator of cellular polyamine levels from yeasts to mammals is the protein antizyme. The antizyme gene consists of two overlapping reading frames with ORF2 in the +1 frame relative to ORF1. A programmed +1 ribosomal frameshift occurs at the last codon of ORF1 and results in the production of full-length antizyme protein. The efficiency of frameshifting is proportional to the concentration of polyamines, thus creating an autoregulatory circuit for controlling polyamine levels. The mRNA recoding signals for frameshifting include an element 5' and a pseudoknot 3' of the shift site. The present work illustrates that the ORF1 stop codon and the 5' element are critical for polyamine sensing, whereas the 3' pseudoknot acts to stimulate frameshifting in a polyamine independent manner. We also demonstrate that polyamines are required to stimulate stop codon readthrough at the MuLV redefinition site required for normal expression of the GagPol precursor protein. PMID- 16269133 TI - Methylation-based molecular margin analysis in hepatocellular carcinoma. AB - The positive surgical margins are associated with postsurgical recurrence in hepatocellular carcinoma patients, and molecular margin analysis is considered more sensitive in detecting preneoplastic lesions than conventional histological margin examination. To evaluate the feasibility of methylation-based molecular margin analysis in HCC and explore its clinical application, we investigated CDKN2A methylation status in the surgical margins of 20 HCC patients using a nested BS-MSP protocol and compared the methylation patterns in resection margins with those in the corresponding tumor and adjacent nonmalignant tissues. The results showed that a considerable frequency (35%, 7 of 20) of CDKN2A methylation was present in histologically negative margins, and methylation pattern analysis might be valuable for studying the cellular origin of recurrent carcinoma. Therefore, methylation-based molecular surgical margin analysis offers a promising tool in prognosis for HCC patients who underwent hepatectomy. PMID- 16269134 TI - Role of the conserved threonine 309 in mechanism of oxidation by cytochrome P450 2D6. AB - Based on sequence alignments and homology modeling, threonine 309 in cytochrome P450 2D6 (CYP2D6) is proposed to be the conserved I-helix threonine, which is supposed to be involved in dioxygen activation by CYPs. The T309V mutant of CYP2D6 displayed a strong shift from O-dealkylation to N-dealkylation reactions in oxidation of dextromethorphan and 3,4-methylenedioxymethylamphetamine. This may be explained by an elevated ratio of hydroperoxo-iron to oxenoid-iron of the oxygenating species. In consistence, using cumene hydroperoxide, which directly forms the oxenoid-iron, the T309V mutant again selectively catalyzed the O dealkylation reactions. The changed ratio of oxygenating species can also explain the decreased activity and changed regioselectivity that were observed in 7 methoxy-4-(aminomethyl)-coumarin and bufuralol oxidation, respectively, by the T309V mutant. Interestingly, the T309V mutant always showed a significantly increased, up to 75-fold, higher activity compared to that of the wild-type when using cumene hydroperoxide. These results indicate that T309 in CYP2D6 is involved in maintaining the balance of multiple oxygenating species and thus influences substrate and regioselectivity. PMID- 16269135 TI - Alterations in cellular phenotypes differentiating from embryonic rat brain neurosphere cultures by immunoselection of neuronal progenitors. AB - The neurosphere culture system is widely used to expand neural stem/progenitor cells in vitro and to provide a source of cells for transplantation approaches to CNS disorders. This study describes the populations of neurones, astrocytes and oligodendrocytes which differentiated from embryonic day (E) 14 rat cortical and striatal tissue grown as neurosphere cultures over three passages. The percentages of cells that adopted neuronal phenotypes decreased with passage, astrocytic percentages increased and oligodendrocytic percentages remained constant. In the second part of this study, immunomagnetic separation was used to positively select neuronal progenitor cells from E14 rat cortical and striatal tissue using an antibody, 2F7, which recognises an epitope on the cell surface of pre- and post-mitotic neurones. These immunomagnetically selected cells were grown as neurosphere cultures over three passages and gave rise to significantly different percentages of neurones, astrocytes and oligodendrocytes than those found in the baseline study. In particular, the percentage of neurones arising from the second and third passages was significantly higher following immunoselection. This indicates that neuronal progenitor cells can be isolated using immunomagnetic separation and then expanded using the neurosphere culture system, to generate enriched populations of neurones that can be used in CNS repair. PMID- 16269136 TI - Capsaicin differentially modulates voltage-activated calcium channel currents in dorsal root ganglion neurones of rats. AB - It is discussed whether capsaicin, an agonist of the pain mediating TRPV1 receptor, decreases or increases voltage-activated calcium channel (VACC) currents (I(Ca(V))). I(Ca(V)) were isolated in cultured dorsal root ganglion (DRG) neurones of rats using the whole cell patch clamp method and Ba2+ as charge carrier. In large diameter neurones (>35 micorm), a concentration of 50 microM was needed to reduce I(Ca(V)) (activated by depolarizations to 0 mV) by 80%, while in small diameter neurones (< or =30 microm), the IC50 was 0.36 microM. This effect was concentration dependent with a threshold below 0.025 microM and maximal blockade (>80%) at 5 microM. The current-voltage relation was shifted to the hyperpolarized direction with an increase of the current between -40 and -10 mV and a decrease between 0 and +50 mV. Isolation of L-, N-, and T-type calcium channels resulted in differential effects when 0.1 microM capsaicin was applied. While T-type channel currents were equally reduced over the voltage range, L-type channel currents were additionally shifted to the hyperpolarized direction by 10 to 20 mV. N-type channel currents expressed either a shift (3 cells) or a reduction of the current (4 cells) or both (3 cells). Thus, capsaicin increases I(Ca(V)) at negative and decreases I(Ca(V)) at positive voltages by differentially affecting L-, N-, and T-type calcium channels. These effects of capsaicin on different VACCs in small DRG neurones, which most likely express the TRPV1 receptor, may represent another mechanism of action of the pungent substance capsaicin in addition to opening of TRPV1. PMID- 16269137 TI - Clorazepate affects cell surface regulation of delta and kappa opioid receptors, thereby altering buprenorphine-induced adaptation in the rat brain. AB - Concomitant abuse of buprenorphine (BPN) and benzodiazepines (BZD) may relate to a pharmacodynamic interaction between the two. The objective of the present work was to investigate the acute and chronic effects of clorazepate (CRZ) alone or in combination with BPN on selective kappa opiate tritiated ligand [3H]-U69 593 and delta opiate radioligand [3H]-deltorphine II binding in the rat brain. Bmax (maximal receptor density) and Kd (the dissociation constant) were directly determined at different brain regions of interest (ROI) selected for high densities of kappa and/or delta receptors in rats treated with BPN and/or CRZ. The agents were administered either once or for 21 consecutive days. Differences in Bmax and Kd (for both specific ligands) were related to drug treatment and receptor location. Globally, single BPN administration induced no changes in kappa or delta opiate receptor binding, whereas repeated BPN administration up regulated kappa receptor density and decreased delta affinity. At the kappa receptor level, repeated administration of CRZ acted only on Kd, whereas the delta receptor was up-regulated. Repeated addition of CRZ to BPN had no effect on kappa receptor Bmax versus chronic controls. By significantly decreasing Bmax, CRZ nullified the effect of chronic BPN on the kappa receptor. The modifications were strongest in the nucleus accumbens, where both types of receptor occur. Treatments had region-selective effects in some brain areas, such as the amygdala, periaqueductal gray matter, hypothalamus and caudate putamen. Increased mu and delta receptor densities would be expected to provide reinforcement by enhancing reward, and impairment of kappa receptor availability would be expected to decrease aversion. The effects described are likely to influence addictive behavior among people abusing BZD and BPN. PMID- 16269138 TI - Reduced immunoreactivities of a vasoactive intestinal peptide and pituitary adenylate cyclase-activating polypeptide receptor (VPAC1 receptor) in the cerebral cortex, hippocampal region, and amygdala of aged rats. AB - In this study, we examined expressional changes of VPAC1 receptor in aged rat brains using an immunohistochemical approach and found that its immunoreactivities are significantly reduced in the cerebral cortex, hippocampal region, and amygdala of aged rats. These results suggest that this reduction could underlie aging-associated memory/learning deficits and several other age induced functional changes in these areas. However, the functional consequences of these down-regulations require further elucidation. PMID- 16269139 TI - Combined use of a cytoprotectant and rehabilitation therapy after severe intracerebral hemorrhage in rats. AB - After moderate intracerebral hemorrhage (ICH), both hypothermia (HYPO) and constraint-induced movement therapy (CIMT) improve recovery and reduce the volume of brain injury. We tested the hypothesis that more severe ICH requires both cytoprotection and rehabilitation to significantly improve recovery. Rats were subjected to a unilateral striatal ICH via collagenase infusion. Rats remained normothermic or were subjected to mild HYPO ( approximately 2 days) starting 12 h later. Fourteen days after ICH, half of the rats received CIMT (7 days of restraint of the less affected limb plus daily exercises); the remainder were untreated. Walking, limb use and skilled reaching were assessed up to 60 days, at which time animals were euthanized and the volume of tissue lost was determined. The HYPO treatment alone did not improve outcome, whereas CIMT alone provided significant benefit on the limb use asymmetry test. In the staircase test, the greatest benefit was achieved with the combination of HYPO and CIMT treatments. The volume of tissue lost after ICH was similar among groups arguing against cytoprotection as a mechanism of functional recovery. Finally, these findings suggest that, at least under the present circumstances (e.g., severe striatal ICH), CIMT provides superior benefit to HYPO and that combination therapy will sometimes further improve recovery. PMID- 16269140 TI - Purification of transgenic plant-derived recombinant human acetylcholinesterase R. AB - Nicotiana benthamiana plants were engineered to express a codon-optimized gene encoding the human acetylcholinesterase-R (AChE) isoform. The transgenic plants expressed the protein at >0.4% of total soluble protein, and the plant-produced enzyme was purified to homogeneity. Following lysis, procainamide affinity chromatography and anion-exchange chromatography, more than 400-fold purification was achieved and electrophoretic purity was obtained. This pure protein is kinetically indistinguishable from the only commercially available source of human acetylcholinesterase, which is produced in mammalian cell culture. Thus, we have demonstrated a model system for the production of acetylcholinesterase, which is not susceptible to the quantitative limitations or mammalian pathogens associated with purification from mammalian cell culture or human serum. PMID- 16269141 TI - Involvement of oxidative stress in the pre-malignant and malignant states of cervical cancer in women. AB - OBJECTIVES: To evaluate the potential role of oxidative stress in the evolution of cervical cancer, including its pre-malignant states. DESIGN AND METHODS: Erythrocytes thiobarbituric acid reactive substances (TBARS) levels, plasma vitamin C and thiol content and total blood delta-ALA-D levels were estimated in 46 untreated cervical cancer and pre-malignant patients and in 46 age-sex-matched controls. RESULTS: Erythrocytes from patients, regardless of disease state, pre malignant (low squamous intraepithelial lesion--LSIL and high squamous intraepithelial lesion--HSIL) or cancer, showed a significant 2-3 times increase in TBARS levels (P<0.01). Plasma vitamin C was lower in the carcinoma group (P<0.01). The reactivation index of delta-aminolevulinate dehydratase (delta-ALA D) was higher in the patient group, when compared to control (P<0.01). CONCLUSION: LSIL, HSIL or cervical cancer can be associated with changes in 3 indicators of oxidative stress: increase in erythrocyte TBARS, ALA-D reactivation index and a decrease in vitamin C content, that may play an important role in carcinogenesis. PMID- 16269142 TI - Sex- and segment-specific modulation of gene expression profiles in Drosophila. AB - Homeotic and sex-determining genes control a wide range of morphological traits by regulating the expression of different target genes in different tissues. The identity of most of these target genes remains unknown, and it is not even clear what fraction of the genome is regulated in a segment- and sex-specific manner. In this report, we examine segment- and sex-specific gene expression in Drosophila pupal legs. The first and second legs in Drosophila have clearly distinguishable bristle patterns. Bristle pattern in the first leg also differs between males and females, whereas the second leg has no overt sexual dimorphism. To identify the genes responsible for these differences, we compared transcriptional profiles between male and female first and second legs during early pupal development. The extent of sexually dimorphic gene expression parallels morphological differences: over 100 genes are expressed sex specifically in the first leg, whereas no sexual differences are seen in the second leg. Segmental differences are less extensive than sexual dimorphism and involve fewer than 14 genes. We have identified a novel gene, CG13857, that is expressed exclusively in the first leg in a pattern that suggests this gene may play an important role in specifying segment- and sex-specific bristle patterns. PMID- 16269143 TI - Ca2+-dependent K+ channels are targets for bradykinin B1 receptor ligands and for lipopolysaccharide in the rat aorta. AB - Although rat aorta smooth muscle cells in culture constitutively express bradykinin B1 receptors, the normotensive rat aorta does not respond to the bradykinin B1 receptor agonist des-Arg9-bradykinin, whereas vessels from the spontaneously hypertensive rat (SHR) respond to bradykinin B1 receptor agonists with cell membrane hyperpolarization and relaxation. Bacterial lipopolysaccharide also is inactive on the normotensive rat but hyperpolarizes the SHR aorta. To determine whether this could be due to the increased intracellular Ca2+ concentration ([Ca2+]i) in the SHR, we raised [Ca2+]i in normotensive rats by treatment with thapsigargin. In the thapsigargin-treated aorta, both lipopolysaccharide and des-Arg9-bradykinin induced hyperpolarization, which was reversed by the Ca2+-dependent K+ channel inhibitor iberiotoxin and by the bradykinin B1 receptor antagonists Lys-[Leu8]-des-Arg9-bradykinin and [Leu8]-des Arg9-bradykinin. Thus the bradykinin B1 receptor, as well as lipopolysaccharide, needs activated Ca2+-dependent K+ channels for functional expression. The two bradykinin B1 receptor inhibitors, however, have effects on Ca2+-dependent K+ channels which are not mediated by bradykinin B1 receptors. PMID- 16269144 TI - Effect of pravastatin, simvastatin and atorvastatin on the phagocytic activity of mouse peritoneal macrophages. AB - Since cholesterol and lipid content may affect cell membrane fluidity, we assumed that treatment of mice with lipid lowering statins would enhance the engulfing capacity of their macrophages. Four groups of animals were examined. Group I treated with pravastatin, group II--with simvastatin--both drugs in a dosage of 40 mg/kg daily, 5 days/week for a total of 3 weeks. Mice in group III received atorvastatin 5 mg/kg for the same time period. Group IV--untreated animals serving as controls. The phagocytic capacity of the peritoneal macrophages was evaluated by their ability to engulf latex particles. In addition, the mitogen response of the peripheral blood mononuclear cells (PBMC) and splenocytes to Con A and PHA was examined. Compared to the controls, the percentage of phagocyting cells in pravastatin-treated mice was enhanced by 18%, with simvastatin--by 24% and in atorvastatin-treated animals by 8%. The three statins increased the phagocytic index by 79.5%, 88.8% and 62%, respectively. The mitogen response of splenocytes from mice treated with the three statins to Con A increased by 68%, 48% and by 40%, respectively. Compared with the controls the response to PHA was higher in animals treated with pravastatin (84%), simvastatin (73%) and atorvastatin (57%). The response of PBMC from statin-treated animals to both mitogens did not differ from that of the controls. The results suggest that statins, at least those hereby investigated, may exert a beneficial effect on the immune function of the macrophages. PMID- 16269146 TI - Attitudes toward the discontinuation of female genital cutting among men and women in Guinea. AB - OBJECTIVE: To investigate socioeconomic correlates of and gender differences in attitudinal support for the discontinuation of FGC in Guinea. METHOD: Data from structured interviews of men aged 15-59 and women aged 15-49 years in the 1999 Demographic and Health Survey and multiple logistic regression methods were used to examine the relationship of socioeconomic factors and gender to attitudinal support for the discontinuation of FGC. RESULT: More than 9 out of 10 women had undergone FGC. Attitudinal support for FGC discontinuation was more prevalent among men than women. The odds of supporting the discontinuation of FGC were negatively related to beliefs in social approval of and religious support for FGC and its enhancement of women's marriageability, the number of perceived advantages of FGC, and women's low socioeconomic status. CONCLUSION: Community education, improvements in women's socioeconomic status and traditional and religious leader involvement would be critical for FGC eradication. PMID- 16269145 TI - Vesicular monoamine transporter-2 and aromatic L-amino acid decarboxylase gene therapy prevents development of motor complications in parkinsonian rats after chronic intermittent L-3,4-dihydroxyphenylalanine administration. AB - Motor complications after chronic L-3,4-dihydroxyphenylalanine (L-DOPA) therapy occur partly because of the sensitization to dopaminergic agents resulting from pulsatile dopaminergic stimulation. The loss of presynaptic storage contributes to short duration of action by dopamine. Vesicular monoamine transporter-2 (VMAT 2) controls intraneuronal dopamine storage by packaging dopamine into synaptic vesicles, thereby allowing exocytotic release of dopamine. Using primary fibroblast doubly transduced with VMAT-2 and aromatic L-amino acid decarboxylase (AADC) genes, we previously demonstrated the beneficial effects of such double gene transduction in the production, storage, and gradual release of dopamine in vitro and in vivo. In this study, we further evaluate the effect of achieving sustained level of dopamine within the striata by VMAT-2 gene on behavioral response of parkinsonian rats after chronic intermittent L-DOPA administration. Primary fibroblast (PF) cells were genetically modified with AADC and VMAT-2 genes. We grafted primary fibroblast cells, PF with AADC (PFAADC), or doubly transduced PF with AADC and VMAT-2 (PFVMAA) (n = 6 for each group) into parkinsonian rat striata and administered L-DOPA (25 mg/kg/day) intermittently for 4 weeks. For behavioral study, we employed a model of akinesia using forepaw adjusting steps (FAS) that have been well characterized to reflect the effect of the lesion and the antiparkinsonian effect of dopaminergic drugs and transplants. The duration of FAS response to L-DOPA was sustained for a longer duration in rats grafted with PFVMAA cells than in those grafted with either control cells or cells with AADC alone. In PFVMAA-grafted animals, prolonged duration of FAS responses to L-DOPA was sustained even 6 weeks after discontinuation of 4-week intermittent L-DOPA treatment. These findings suggest that the restoration of dopamine storage capacity could enhance the efficacy of L-DOPA therapy and attenuate the motor fluctuations that result from chronic intermittent L-DOPA administration. The gene therapy expressing AADC and VMAT-2 along with systemic L DOPA therapy could provide a novel treatment strategy to prevent motor fluctuations. PMID- 16269148 TI - Affinity capillary electrophoresis of DNA for detection of single-nucleotide polymorphisms and point mutations. Comprehensive study for optimization of the weak affinity. AB - A single-stranded DNA and its point mutant can be separated with affinity capillary electrophoresis (ACE) in which an appropriate ligand DNA is used as a pseudo-stationary affinity phase. In this paper, we systematically examine the effects of ligand sequence, capillary temperature, and cation concentration on the ACE separation quality, which is quantitatively evaluated in terms of resolution and peak height. For fine tuning of the affinity, control of MgCl(2) concentration and insertion of a spacer sequence into the ligand DNA are more effective than control of the capillary temperature. For design of the ligand DNA, a simple strategy is proposed, based on the prediction of melting temperature. This strategy was tested with eleven different sample sequences. All of them were separated under the same conditions (250 microM MgCl(2) and 25 degrees C), and nine cases of them had satisfactory separation qualities. PMID- 16269147 TI - Cervicography as an adjunctive test to visual inspection with acetic acid in cervical cancer detection screening. AB - OBJECTIVE: Validate Digital Cervicography as adjunctive cervical cancer screening test to VIA and Cytology. METHOD: Women (1292) were submitted to both cytological and VIA tests to obtain a sample of 301 positive VIA cases (23.3%) which have had the Digital Cervicography (DC) taken and evaluated. Just cases considered positive by DC (149/301 cases--49.5%) and/or positive Pap smear tests were referred to colposcopy and biopsy whenever indicated. RESULTS: Cervical smear was positive in 5.4%, including LGSIL (4%), HGSIL (1%) and one case of invasive cancer (0.4%), and 1.6% of ASCUS or AGUS). DC identified 81 positive cases (74 LGSIL; 5 HGSIL; 2 cancers). The sensitivity of cytology was 14.8%, and the DC was 100%. Corresponding specificities were 95.4% and 69.1%, respectively. The NPV and PPV for DC were 100% and 54.4%. CONCLUSION: DC increases sensitivity, specificity and positive predictive value of VIA, becoming a valid adjunctive screening test for cervical cancer in low resource settings. PMID- 16269149 TI - Determination of mustard and lewisite related compounds in abandoned chemical weapons (Yellow shells) from sources in China and Japan. AB - Knowledge of the states of the contents in chemical munitions that Japanese Imperial Forces abandoned at the end of World War II in Japan and China is gravely lacking. To unearth and recover these chemical weapons and detoxify the contents safely, it is essential to establish analytical procedures to definitely determine the CWA contents. We established such a procedure and applied it to the analysis of chemicals in the abandoned shells. Yellow shells are known to contain sulfur mustard, lewisite, or a mixture of both. Lewisite was analyzed without thiol derivatization, because it and its decomposition products yield the same substances in the derivatization. Analysis using our new procedure showed that both mustard and lewisite remained as the major components after the long abandonment of nearly 60 years. The content of mustard was 43% and that of lewisite 55%. The viscous material found was suggested to be mostly oligomers of mustard. Comparison of the components in the Yellow agents with mustard recovered in both Japan and China showed a difference in the impurities between the CWAs produced by the former Imperial navy and those by the former Imperial army. PMID- 16269150 TI - Gas chromatographic-mass spectrometric analysis of essential oils from Pimpinella aurea, Pimpinella corymbosa, Pimpinella peregrina and Pimpinella puberula gathered from Eastern and Southern Turkey. AB - Essential oils from fruits, stems and leaves and roots of Pimpinella aurea DC., P. corymbosa Boiss., P. peregrina L. were analyzed by gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) techniques. Fruits and aerial parts of P. puberula (DC.) Boiss were also evaluated. A total of 140 different compounds were identified, and significant qualitative and quantitative differences were observed among the samples. In fact, the main constituents of each species were different and only the oils extracted from roots shared the same principal compound, epoxy pseudoisoeugenyl-2-methyl butyrate (26.8-42.8%). The other fractions were dominated by different sesquiterpene compounds although in three of them, P. aurea stem and leaves, P. puberula fruits and P. puberula stems and leaves, monoterpene constituents also appear as main ones. PMID- 16269151 TI - Developments in the application of counter-current chromatography to plant analysis. AB - Counter-current chromatography is a very versatile separation technique which does not require a solid stationary phase. It relies simply on the partition of a sample between the two phases of an immiscible solvent system. Some of the more recent applications of the method to the separation of plant-derived natural products are described here. Crude plant extracts and semi-pure fractions can be chromatographed, with sample loads ranging from milligrams to grams. Aqueous and non-aqueous solvent systems are used and the separation of compounds with a wide range of polarities is possible. The technique is complementary to other chromatographic methods and is compatible with gradient systems. The possibilities for solvent selection are almost limitless but some guidelines for the choice of successful systems are presented. PMID- 16269152 TI - Membrane and extracellular antigens of Paracoccidioides brasiliensis (Mexo): identification of a 28-kDa protein suitable for immunodiagnosis of paracoccidioidomycosis. AB - In this work, we analyzed serological responses of paracoccidioidomycosis (PCM) patients to membrane and extracellular antigens (Mexo) of Paracoccidioides brasiliensis by ELISA, immunoblot technique and immunofluorescence assays to identify a specific antigen profile. Among 140 PCM serum samples analyzed, a homogeneous IgG response to Mexo was observed. The specificity of this antigen was 96.6% in relation to control sera and 81.2% to sera from patients with diverse infections. Patients undergoing treatment for more than 1 year showed a reduced antibody response against Mexo. These results suggest that the presence of anti-Mexo antibodies might be an indicator of active disease. A protein from Mexo with a molecular weight of 28 kDa (Pb28) was the most specific antigen in humoral immune responses to PCM, since it reacted with 100% of patient sera and did not react with heterologous serum samples tested. This protein was purified by molecular filtration chromatography in FPLC system and, when tested by immunoblotting, it maintained its reactivity and specificity of 100% with PCM sera. The Pb28 N-terminal amino acid sequence comparison analysis in the non redundant GenBank database at NCBI revealed no significant homology to known PCM proteins or to other fungal proteins of known function. Since the 28-kDa protein of P. brasiliensis seems to be specific for PCM, it can be used as an alternative antigen in immunoblotting diagnostic methods. PMID- 16269153 TI - Crystal structures of active LytM. AB - Lysostaphin-type enzymes are metalloendopeptidases that are present in bacteriophages and in bacteria. They share the catalytic domain, but normally contain other domains as well. The well-characterized enzymes in this group are all specific for the pentaglycine crosslinks in the cell walls of some Gram positive bacterial species. Lysostaphin-type enzymes are synthesized as secreted preproenzymes and require proteolytic activation for maturation. Although lysostaphin, the prototypical peptidase in the group, is widely used as a tool in biotechnology and developed as an antistaphylococcal agent, the detailed structure of this enzyme is unknown. So far, only one lysostaphin-type enzyme, the Staphylococcus aureus autolysin LytM, has been crystallized in its full length, inactive form. Here, we describe the synthesis of a convenient reporter substrate, characterize the metal and pH-dependence of an active LytM fragment, and present its crystal structure in three crystal forms at different pH values that either support or do not support activity. In all structures, we find an extended, long and narrow groove that has the active site at its bottom and is delineated on the sides by the most flexible regions of the molecule. In two cases, the groove is partially filled by a loop of a neighbouring molecule in the crystal. As the loop contains three consecutive glycine residues, this crystal packing effect supports the interpretation that the groove is the substrate binding cleft. To characterize the substrate-binding mode more closely, a phosphinate analogue of tetraglycine was synthesized. Although tetraglycine is a substrate of the active LytM fragment, the phosphinate analogue turned out to be a very poor inhibitor. Crystals that were grown in its presence contained an L+ tartrate molecule from the crystallization buffer and not the phosphinate in the active site. PMID- 16269154 TI - Characterizing conserved structural contacts by pair-wise relative contacts and relative packing groups. AB - To adequately deal with the inherent complexity of interactions between protein side-chains, we develop and describe here a novel method for characterizing protein packing within a fold family. Instead of approaching side-chain interactions absolutely from one residue to another, we instead consider the relative interactions of contacting residue pairs. The basic element, the pair wise relative contact, is constructed from a sequence alignment and contact analysis of a set of structures and consists of a cluster of similarly oriented, interacting, side-chain pairs. To demonstrate this construct's usefulness in analyzing protein structure, we used the pair-wise relative contacts to analyze two sets of protein structures as defined by SCOP: the diverse globin-like superfamily (126 structures) and the more uniform heme binding globin family (a 94 structure subset of the globin-like superfamily). The superfamily structure set produced 1266 unique pair-wise relative contacts, whereas the family structure subset gave 1001 unique pair-wise relative contacts. For both sets, we show that these constructs can be used to accurately and automatically differentiate between fold classes. Furthermore, these pair-wise relative contacts correlate well with sequence identity and thus provide a direct relationship between changes in sequence and changes in structure. To capture the complexity of protein packing, these pair-wise relative contacts can be superimposed around a single residue to create a multi-body construct called a relative packing group. Construction of convex hulls around the individual packing groups provides a measure of the variation in packing around a residue and defines an approximate volume of space occupied by the groups interacting with a residue. We find that these relative packing groups are useful in understanding the structural quality of sequence or structure alignments. Moreover, they provide context to calculate a value for structural randomness, which is important in properly assessing the quality of a structural alignment. The results of this study provide the framework for future analysis for correlating sequence changes to specific structure changes. PMID- 16269155 TI - Isolated cerebral mucormycosis: report of a case and review of the literature. AB - Isolated cerebral mucormycosis is a rare but life-threatening infection that generally occurs in patients with intravenous drug abuse or immune deficiency. We report a case of primary cerebral mucormycosis in a healthy adult. Whole body autopsy in this case revealed cerebral mucormycosis with prominent vascular pathology and hemorrhagic necrosis. No nasal sinus, orbital or other primary locus of fungus infection was discovered. Review of the previously reported 30 cases of isolated cerebral mucormycosis revealed associated systemic predisposition in 11 patients and history of intravenous drug abuse in 17 cases. In the remaining two cases, the diagnosis of fungal infection was made only after surgical exploration. Early tissue diagnosis and the consequent surgical excision of the necrotic tissue and aggressive antifungal therapy might salvage life in this fatal condition. PMID- 16269156 TI - Ganoderma lucidum polysaccharides peptide inhibits the growth of vascular endothelial cell and the induction of VEGF in human lung cancer cell. AB - Ganoderma lucidum Polysaccharide Peptide (Gl-PP) has shown some effects as anti tumors in mice and potential anti-angiogenesis. In this study, we elucidated the possible mechanism of Gl-PP action on anti-angiogenesis of tumor. Our research indicated that the proliferation of HUVECs was inhibited by Gl-PP in a dose dependent fashion, but not because of cytotoxicity. Flow cytometric studies revealed that Gl-PP treatment of HUVECs could induce cell apoptosis directly. Moreover, addition of Gl-PP also led to a reduction of Bcl-2 anti-apoptotic protein expression and an increase of Bax pro-apoptotic protein expression of HUVECs. Therefore, inducing cell apoptosis by Gl-PP might be the mechanism of inhibiting HUVEC proliferation. Human lung carcinoma cells PG when exposed to high dose of Gl-PP in hypoxia for 18 h resulted in a decrease in the secreted VEGF. Taken together, these findings support the hypothesis that the key attribute of the anti-angiogenic potential of Gl-PP is that it may directly inhibit vascular endothelial cell proliferation or indirectly decrease growth factor expression of tumor cells. PMID- 16269157 TI - Vasodilatory and anti-inflammatory effects of the aqueous extract of rhubarb via a NO-cGMP pathway. AB - While conducting an in vitro screen of various medicinal plant extracts, an aqueous extract of rhubarb (Rheum undulatum L, AR) was found to exhibit a distinct vasorelaxant activity. AR induced a concentration-dependent relaxation of the phenylephrine-precontracted aorta. This effect disappeared with the removal of functional endothelium. Pretreatment of the aortic tissues with N(G) nitro-L-arginine methyl ester (L-NAME), methylene blue, or 1H-[1,2,4]-oxadiazole [4,3-alpha]-quinoxalin-1-one (ODQ) inhibited the relaxation induced by AR. Incubation of human umbilical vein endothelial cells (HUVECs) with AR increased the production of cGMP in a dose-dependent manner, but this effect was blocked by pretreatment with L-NAME and ODQ, respectively. AR treatment attenuated TNF-alpha induced NF-kappaB p65 translocation in HUVECs in a dose-dependent manner. In addition, AR suppressed the expression levels of adhesion molecules including ICAM-1 and VCAM-1 induced by TNF-alpha in HUVECs. TNF-alpha-induced MCP-1 expression was also attenuated by the addition of AR. This attenuation was blocked by pretreatment with either L-NAME or ODQ. AR treatment inhibited cellular adhesion of U937 cells onto HUVECs induced by TNF-alpha. Taken together, the present study suggests that AR dilates vascular smooth muscle and suppresses the vascular inflammatory process via endothelium-dependent NO/cGMP signaling. PMID- 16269159 TI - Evaluating compliance with national guidelines for the clinical, laboratory and public health management of tuberculosis in a low-prevalence English district. PMID- 16269158 TI - Area characteristics and determinants of hospitalised childhood burn injury: a study in the city of Cape Town. AB - OBJECTIVES: To investigate the impact of local living circumstances on the level of childhood burn injuries. STUDY DESIGN/METHOD: This study was an ecological investigation encompassing the 634 residential areas of the city of Cape Town (about 668,900 children aged 12 years and younger). Fifteen socio-physical features of these areas that describe the population, household, dwelling types and services were considered using data from the 2001 census. Data of childhood burn injuries were gathered from the Red Cross Children's Hospital's register over 1999-2000 (n=923). Area features were synthesized into three main dimensions using factor analysis (principal axis method). Each dimension was split into three (exposure) levels, and the effect of each dimension on childhood burn injury was measured, compiling odds ratios with 95% confidence intervals and using the most favourable exposure level as the reference category within each dimension. RESULTS: The main dimensions of contextual exposure were defined as housing conditions (five variables), child dependency (three variables) and socio economic barriers (three variables). Each aspect had a significant impact on the risk of burn injury, with increased levels of exposure leading to increased risk of burns. For housing conditions (Factor 1) and socio-economic barriers (Factor 3), a graded relationship was observed with noteably increased odds for increased levels of exposure. CONCLUSIONS: The occurrence of child burn injury is highly influenced by various features of the local environment. These features may be essential targets for sustainable childhood burn injury control and prevention programmes. PMID- 16269160 TI - Effect of healthcare on mortality: trends in avoidable mortality in Australia and comparisons with Western Europe. AB - OBJECTIVES: Using the concept of avoidable mortality, international studies suggest that healthcare has been effective in reducing mortality. This paper provides an analysis of avoidable mortality in Australia and compares trends with those of Western Europe. METHODS: Using unit-record mortality data, we calculated avoidable mortality rates in Australia for 1968-2001. We partitioned avoidable causes into three categories: those amenable to medical care; those mainly responsive to health policy; and ischaemic heart disease. We used Poisson regression to model the trends. We compared trends with those of nine European countries using published data. RESULTS: Total avoidable death rates fell by 68% in females and 72% in males. The corresponding non-avoidable death rates fell by 35 and 33%. The annual declines in avoidable mortality rates were: 3.47% [95% confidence intervals (CI) 3.44-3.50%] in males and 3.89% (95% CI 3.86-3.91%) in females. For non-avoidable mortality rates, the annual declines were 1.09% (95% CI 1.05-1.13%) and 0.95% (95% CI 0.92-0.98%), respectively. In females, declines in death rates from causes amenable to medical care contributed 54% to the decline in avoidable mortality rates, ischaemic heart disease contributed 45%, and causes responsive to health policy intervention contributed 1%. In males, the corresponding contributions were 32, 57 and 11%. These rates, and the declines between 1980 and 1998, were comparable with selected European countries, with Australia's ranking improving over the period. CONCLUSION: Trends in avoidable mortality in Australia suggest that the Australian healthcare system has been effective in improving population health. Australia's experience compares favourably with that of Europe. PMID- 16269161 TI - History of breastfeeding and Helicobacter pylori infection in children: results of a community-based study from northeastern Brazil. AB - The aim of this study was to evaluate the role of breastfeeding and the infection status of the mother in the acquisition of Helicobacter pylori infection in a poor urban community in northeastern Brazil. Helicobacter pylori status was evaluated by 13C-urea breath test in individuals under the age of 14 years and by ELISA in the mothers. The prevalence of H. pylori infection was 55.8% (197/353) in the children and it increased with age (P<0.0001). Of the children in whom breastfeeding status was known, 93.2% (316/339) were breastfed. The H. pylori prevalence did not differ between breastfed and never breastfed children (55% vs. 52%) even when children were breastfed for >6 months. The prevalence of infection was much higher in children whose mothers were H. pylori infected than in children whose mothers were not infected, resulting in a crude odds ratio (OR) of 3.11 (95% CI 1.57-6.19) and 2.40 after adjustment for potential confounders (95% CI 1.12-5.15). This study suggests that breastfeeding does not protect against acquisition of H. pylori in northeastern Brazil; conversely, an infected mother may have an important role in transmission of the disease to the child. PMID- 16269162 TI - Individual venom variability in Crotalus durissus ruruima snakes, a subspecies of Crotalus durissus from the Amazonian region. AB - Venoms of six specimens of Crotalus durissus ruruima snakes from the same geographical site in the Brazilian State of Roraima, were individually assayed for their main pharmacological properties. Quantitative and qualitative differences were found and the presence of crotoxin-like isoforms in these venoms was indicated. Our findings corroborate the existence of a considerable intrapopulational variability in C. d. ruruima venoms, and the importance of using a pool of venoms for antivenom production, in general, in order to assure the neutralization of the maximum possible number of toxins from a given species. PMID- 16269163 TI - Envenoming by the viperid snake Eristicophis macmahonii. AB - Little is known about the effects of bites by the rare viperid snake Eristicophis macmahonii. A 31-year-old Belgian man, who keeps these animals as a hobby, was bitten on a finger by one of his pets while trying to give it some medicine. The patient developed syncope, followed by a sudden extensive skin rash and massive edema of the arm. He recovered completely, except for a small area of limited tissue necrosis on the bitten finger. PMID- 16269164 TI - Crystal structure of a novel phospholipase A2 from Naja naja sagittifera with a strong anticoagulant activity. AB - This is the first PLA(2) crystal structure from group I that shows a strong anticoagulant property. The monomeric PLA(2) was purified from the venom of Naja naja sagittifera (Indian cobra). Its amino acid sequence has been determined using cDNA technique. The amino acid sequence of sPLA(2) contains three positively charged and two negatively charged residues in the segment 54-71 (numbering scheme of sPLA(2)) thus giving this region an overall cationic amphiphilic surface. This suggested the presence of an anticoagulant activity in sPLA(2). The enzyme was crystallized using hanging drop vapour diffusion method in the presence of calcium chloride. The crystals belong to space group P4(1) with cell dimensions of a=b=42.0A, c=65.9A. The X-ray crystal structure was determined at 1.8A resolution using molecular replacement method and refined to an R value of 0.179 for 10,023 reflections. The overall scaffolding of sPLA(2) is essentially similar to those observed for other group I PLA(2)s. However, the conformations of various surface loops were found to be significantly different. The most significant observation pertains to the anticoagulant loop in which both the acidic residues are engaged in intramolecular interactions whereas all the three basic residues are free to interact with other molecules. This makes the sPLA(2) a potentially strong anticoagulating molecule. PMID- 16269165 TI - Catalpol prevents the loss of CA1 hippocampal neurons and reduces working errors in gerbils after ischemia-reperfusion injury. AB - Catalpol, an iridoid glycoside, contained richly in the roots of Rehmannia glutinosa, was found for the first time to be of neuroprotection in gerbils subjected to transient global cerebral ischemia. Catalpol (1 mg/kg ip) used immediately after reperfusion and repeatedly at 12, 24, 48 and 72 h significantly rescued neurons in hippocampal CA1 subfield and reduced working errors during behavioral testing. The neuroprotective efficacy of catalpol became more evident when the doses of catalpol were increased to 5 and 10mg/kg. In addition, it was exciting that the significant neuroprotection by catalpol was also evident when catalpol was applied up to 3 h after ischemia. But the neuroprotective efficacy of catalpol became weak when catalpol was given at 6h after ischemia. Of great encouragement was the finding that the neuroprotection of catalpol could be seen not only in a short post-ischemic period (12 days) but also in a long period (35 days). All these indicated that catalpol was truly neuroprotective rather than simply delayed the onset of neuronal damage and might be of therapeutic value for the treatment of global cerebral ischemia. PMID- 16269167 TI - Immunohistochemical study of proliferating cell nuclear antigen (PCNA) in duckling liver fed with aflatoxin B1 and esterified glucomannan. AB - The effect of esterified glucomannan on aflatoxin B(1) toxicity in ducklings was studied by immunohistochemical staining of proliferating cell nuclear antigen (PCNA) in hepatic cells on formalin-fixed paraffin-embedded liver samples. Cherry Valley ducklings were divided into five groups, 20 birds in each. One of the groups was fed with conventional feed, and the other groups were fed with diet containing 100 ppb aflatoxin B(1), that containing 0.05% esterified glucomannan, or that containing 100 ppb aflatoxin B(1) supplemented with 0.05 or 0.1% esterified glucomannan, from five days of age for one month, and subsequently all the groups were fed with conventional feed for 20 days. Four birds of each group were sacrificed on the 30th, 35th, 40th, 45th and 50th day of feeding, and PCNA on the liver tissue sections was quantitatively analyzed by immunohistochemical staining. The percentage of PCNA-positive hepatocytes was significantly higher in the group given diet containing aflatoxin B(1) than in the other groups, which were not significantly different from each other. The results demonstrate that supplementation of feed with esterified glucomannan is effective in reduction of aflatoxin B(1)-induced hepatic injury in ducklings. PMID- 16269166 TI - Responses of antioxidant system in Arabidopsis thaliana suspension cells to the toxicity of microcystin-RR. AB - Microcystins are cyclic heptapeptide hepatoxins produced by many species of cyanobacteria. The toxic effects and mechanism of microcystins on animals have been well studied both in vivo and in vitro. It was also reported that microcystins had adverse effects on plants. However, to our knowledge, there is no information about the toxic effects and mechanism of microcystins on plant suspension cells. In this study, Arabidopsis thaliana suspension cells were exposed to a range dose of microcystin-RR. Lipid peroxidation, a main manifestation of oxidative damage, was studied and a time- and dose-dependent increase in malondiadehyde was observed. In contrast, glutathione (GSH) levels in the cells decreased after 48 h treatment with 1 and 5mg/L of microcystin-RR. The activities of superoxide dismutase (SOD) and catalase (CAT) increased significantly after 48 h exposure to 1 and 5mg/L of microcystin-RR, but glutathione S-transferase (GST) activity showed no difference compared with the control. These results clearly indicate that microcystin-RR is able to cause oxidative damage in A. thaliana suspension cells. Decrease of GSH content and increases of SOD and CAT activities reveal that the antioxidant system may play an important role in eliminating or alleviating the toxicity of microcystin-RR. The possible toxicity mechanism of microcystin-RR on the A. thaliana suspension cells is also discussed in this paper. PMID- 16269168 TI - Labile aluminium chemistry downstream a limestone treated lake and an acid tributary: effects of warm winters and extreme rainstorms. AB - The outlet from the limestone treated Lake Terjevann consisted mainly of well mixed lake water (mean pH 6.1) during the ice-free seasons including the unusually warm winters of 1992 and 1993. However, during the ice-covered period acidic water (mean pH 4.8, mean inorganic aluminium (Al(i)) about 160 microg/l) from the catchment draining under the lake ice dominated. A downstream tributary was generally acid and rich in aluminium (mean pH 4.6, Al(i) about 230 microg/l). After an extreme rainstorm loaded with sea-salts cation exchange in the soil resulted in more than a doubling of the Al(i) concentration (reaching about 500 microg/l). It took 3-4 months until the Al(i) concentration returned to pre-event levels. During the ice-covered period, the acidic outlet and tributary waters resulted in acidic conditions below the confluence (pH<4.8, Al(i) about 150 microg/l) while during the ice-free periods the more neutral outlet water resulted in higher pH and lower Al(i) concentrations (pH>5.2, Al(i) about 95 microg/l). However, during the latter climatic conditions the water was most probably more harmful to fish due to hydrolysing and polymerizing aluminium. After the sea-salt event, the increased Al(i) concentration in the tributary made the zone below the confluence potentially more toxic (pH approximately 5, Al(i) approximately 250 microg/l). Expected global warming resulting in winter mean temperatures above 0 degrees C may eliminate the seasonal acidification of the outlet from limestone-treated lakes creating permanent toxic mixing zones in the confluence below acidic aluminium-rich tributaries. Besides, more frequent rainstorms as a consequence of global warming may increase the frequency of sea salt events and the Al(i) concentrations in the mixing zones. PMID- 16269169 TI - Pediatric edition thrombosis research. PMID- 16269170 TI - The problem of "lumping versus splitting". PMID- 16269172 TI - The role of hybrid PET/CT in the evaluation of patients with cervical cancer. AB - OBJECTIVE: To assess the role of hybrid PET/CT in the evaluation of patients with cervical cancer. METHODS: 75 patients divided into 3 groups. Group 1 consisted of 16 patients prior to radical surgery. Group 2 consisted of 31 patients prior to pelvic radiotherapy. Group 3 had 28 patients who underwent the examination secondary to suspected recurrent disease. Whole body PET and CT were performed respectively on the same device 1 h after injection of 10 mCi FDG. PET/CT results were correlated to histological, radiological and clinical follow-up data. Only women with >6 months follow-up were included. RESULTS: In 33 patients, pathohistological examinations for extra cervical lesions were obtained. Correlation with PET/CT examination revealed levels of sensitivity (60%), specificity (94%), positive (90%) and negative predictive values (74%). The examination indicated 21 patients with extrapelvic and/or metastatic disease. The follow-up data of this group revealed that 20 patients either died or were alive with active disease, and only one patient was in clinical remission. PET/CT yielded an improved diagnosis for both PET and CT in 43% of the cases by providing better localization and definition of abnormal FDG uptake. CONCLUSIONS: Hybrid PET/CT was found to be of value for detection of cervical cancer metastases, decision-making and planning of irradiation therapy. Using this modality may reduce unnecessary surgical interventions, help modify radiation fields and change therapeutic approaches. Detection of advanced diseases on PET/CT correlates with poor prognosis. However, this examination is less accurate in detecting microscopic diseases and lesions smaller than 1.5 cm. PMID- 16269173 TI - Lead-induced catalase activity differentially modulates behaviors induced by short-chain alcohols. AB - Acute lead administration produces a transient increase in brain catalase activity. This effect of lead has been used to assess the involvement of brain ethanol metabolism, and therefore centrally formed acetaldehyde, in the behavioral actions of ethanol. In mice, catalase is involved in ethanol and methanol metabolism, but not in the metabolism of other alcohols such as 1 propanol or tert-butanol. In the present study, we assessed the specificity of the effects of lead acetate on catalase-mediated metabolism of alcohols, and the ability of lead to modulate the locomotion and loss of the righting reflex (LRR) induced by 4 different short-chain alcohols. Animals were pretreated i.p. with lead acetate (100 mg/kg) or saline, and 7 days later were injected i.p. with ethanol (2.5 or 4.5 g/kg), methanol (2.5 or 6.0 g/kg), 1-propanol (0.5 or 2.5 g/kg) or tert-butanol (0.5 or 2.0 g/kg) for locomotion and LRR, respectively. Locomotion induced by ethanol was significantly potentiated in lead-treated mice, while methanol-induced locomotion was reduced by lead treatment. The loss of righting reflex induced by ethanol was shorter in lead-treated mice, and lead produced the opposite effect in methanol-treated mice. There was no effect of lead on 1-propanol or tert-butanol-induced behaviors. Lead treatment was effective in inducing catalase activity and protein both in liver and brain. These results support the hypothesis that the effects of lead treatment on ethanol-induced behaviors are related to changes in catalase activity, rather than some nonspecific effect that generalizes to all alcohols. PMID- 16269171 TI - Effects of human papillomavirus type 16 E7 protein on the growth of cervical carcinoma cells and immuno-escape through the TGF-beta1 signaling pathway. AB - OBJECTIVE: E7 is regarded as one of the main oncoproteins of high-risk human papillomaviruses (HPVs). It may affect the transforming growth factor beta 1 (TGF beta1) signaling pathway. In this study, the relationship between HPV-16 infection and the functions of three critical factors of the TGF-beta1/Smads pathway was explored to assess the possible role of E7 in the development of cervical cancer. METHODS: The expression of E7, TGF-beta1, TbetaR-II and Smad4 was detected by immunohistochemistry in paraffin-embedded cervical samples, and by RT-PCR and Western blotting in cervical cancer cell lines. The effect of TGF beta1 on the growth of cervical cancer cells were tested by methyl thiazolyl tetrazolium (MTT), and the effects of HPV-16 E7 protein on normal and malignant cervical cells were investigated by flow cytometry. RESULTS: During the progression from benign to malignant lesions, the expression levels of TGF-beta1 and Smad4 increased significantly in cervical carcinoma tissues. The expression of TGF-beta1 was positively correlated with E7 expression. In vitro experiments showed that TGF-beta1 could not inhibit the proliferation of several cervical carcinoma cell lines in long-term regulation, but could inhibit immunologic reactions of peripheral blood mononuclear cells (PBMCs). Blocking E7 expression could lower the expression level of TGF-beta1 and induce cells to enter apoptosis. CONCLUSIONS: Our data indicate that HPV-16 E7 protein plays an important role during the development of cervical cancer by immuno-inhibition and stimulation of tumor cell proliferation through the TGF-beta1/Smads signaling pathway. PMID- 16269174 TI - Effect of hormonal stimulation on bovine follicular response and oocyte developmental competence in a commercial operation. AB - The widespread use of ultrasonography and IVF over the past decade has provided new tools to evaluate how follicles and oocytes react to different superstimulatory treatments. This information may be used to redefine actual hormonal stimulations to improve results of MOET programs and/or obtain improved responses from the "so-called" poor responders. This retrospective study examined data collected over a 5-year period involving oocyte collections in a commercial embryo transfer unit to determine the stimulation protocol that was most effective in producing competent cumulus oocyte complexes, and to determine a definition of a low responder. Overall, the population of small antral follicles at the time of follicle ablation was the most important factor affecting results. This pool of small antral follicles was significantly correlated with the number of follicles at oocyte collection, and to the number of viable and transferable embryos produced. Varying the superstimulatory treatments in terms of type of FSH in association with a shorter or longer coasting period did not affect ovarian response or embryonic development rates. Low responders (less than 10% of the animals in this study) were defined as animals with a lower than average follicular response following superstimulation. Low potential animals were defined as donors producing a limited number of embryos because of the limited population of small antral follicles present in the ovaries at initiation of FSH treatment. Embryo transfer practitioners must distinguish between low responders and low potential animals as modifications to the stimulation protocol for the latter group is unlikely to result in a higher number of transferable embryos. PMID- 16269175 TI - Tracing the glycogen cells with protocadherin 12 during mouse placenta development. AB - Among the different trophoblast subtypes of the mouse placenta, the glycogen cells (GC) are one of the trophoblast subtypes that invade the decidua. We previously established that GC specifically expressed protocadherin 12 (PCDH12). In this paper, we investigated the origin of the PCDH12-positive cells and we characterized their fate in the maternal tissues. Our data indicate that they directly originate from the central part of the ectoplacental cone at embryonic day (E) 7.5. PCDH12-positive cells start to accumulate glycogen from E10.5 and the first migrating cells could be observed from this age. Unlike other placental and decidual cells, GC do not express P-cadherin, which may explain their migration properties in this organ. In the decidua, GC settle in the vicinity of the maternal vascular sinuses but do not incorporate in the endothelium. By the end of gestation (E17.5), most GC islets of the decidua enter into a lytic phase and form large lacunae. These lacunae, filled with glycogen, may provide a substantial source of energy at the end of gestation or during delivery. Our data suggest that spongiotrophoblasts and GC are two independent lineages and we bring insights into GC migration and fate. PMID- 16269176 TI - Neurochemical, neuroautonomic and neuropharmacological acute effects of sibutramine in healthy subjects. AB - Sibutramine is a neuropharmacological drug that exerts central (CNS) and peripheral effects including noradrenaline (NA), and serotonin (5-HT) uptake inhibition. In addition, the drug is able to induce release from DA axons. We measured levels of circulating neurotransmitters in 20 healthy subjects during supine-resting (fasting) state before and after 15 mg of oral sibutramine. Systolic blood pressure (SBP), diastolic blood pressure (DBP), and heart rate (HR) were also monitored. Sibutramine triggered sustained and progressive increase of NA, NA/Ad ratio and DBP. Slight increases of DA were also registered between the 60 and 240 min periods. The rise in DA tended to fade progressively, reaching basal level at 360 min period. Diastolic blood pressure, but neither SBP nor HR, showed significant increases that correlated positively with NA/Ad ratios. Slight but significant negative correlation was also found between DBP and DA. This correlation tended to fade throughout the trial to show no significance at the 360 min period. Although neither plasma serotonin (f-5HT) nor platelet serotonin (p-5HT) values showed significant variation throughout the trial, the f-5HT/p-5HT ratio showed significant decrease throughout. Significant negative correlation was found between f-5HT/p-5HT ratio and NA/Ad ratio. Our results indicate that sibutramine stimulates neural sympathetic activity but not adrenal sympathetic activity in healthy individuals. Further, sibutramine lowers parasympathetic activity. The moderate rise in diastolic blood pressure triggered by sibutramine would be associated with CNS-NA enhancement plus parasympathetic inhibition. PMID- 16269177 TI - Multidrug-resistant Escherichia coli clonal groups causing community-acquired bloodstream infections. AB - OBJECTIVES: A multidrug-resistant Escherichia coli clonal group (designated CgA) has been isolated from women with cystitis and pyelonephritis in several communities. This study was designed to determine if CgA can cause community acquired bloodstream infections. METHODS: All community-acquired bloodstream infections caused by E. coli identified at the San Francisco General Hospital between May 2001 and May 2003 were included. The diagnosis of septicemia was based on admission diagnosis. E. coli isolates were characterized by antibiotic susceptibility profile, enterobacterial repetitive intergenic consensus (ERIC2) PCR, serogrouping, and pulsed field gel electrophoresis (PFGE). RESULTS: A total of 127 individuals with a community-acquired bloodstream infection were identified; 48 (39%) were trimethoprim-sulfamethoxazole (SXT)-resistant. CgA, as defined by ERIC2 PCR, was responsible for 19 (15%) of these infections. Infection with a CgA isolate was associated with an admission diagnosis of cystitis or pyelonephritis (p=0.01). By PFGE, none of the CgA isolates were indistinguishable to the prototype cystitis strain; however, nine bloodstream isolates differed by fewer than six bands. CONCLUSIONS: CgA can cause community-acquired bloodstream infections, but does not appear to cause a disproportionate number of severe extraintestinal infections. This study provides evidence that UTI-causing clonal groups can cause a wide spectrum of disease and are an important clinical and public health concern. PMID- 16269178 TI - Hypoglycemia-induced anoxic brain injury possibly associated with levofloxacin. AB - Reports linking alterations in blood glucose concentrations with fluoroquinolones, mostly gatifloxacin and ciprofloxacin, have been published. We describe an elderly, non-diabetic patient with steroid-induced hyperglycemia prescribed glyburide who later developed severe hypoglycemia resulting in anoxic brain injury soon after initiating therapy with levofloxacin. PMID- 16269179 TI - Short course of prednisolone in patients with solitary cysticercus granuloma: a double blind placebo controlled study. AB - The study was conducted to confirm the role of a short course of oral corticosteroids in patients with solitary cysticercus granuloma with seizures by a double-blind placebo-controlled study. In an open-label trial we, in past, had demonstrated a beneficial role of prednisolone. A short course of prednisolone helped in early resolution of solitary cysticercus granuloma. In this double blind placebo-controlled randomized study, 60 patients with new-onset seizures and a single enhancing computed tomography (CT)-detected lesion of cysticercosis were randomly divided in two groups to receive either anti-epileptic monotherapy with prednisolone (n=30) or anti-epileptic monotherapy along with placebo (n=30). The patients received prednisolone, 1mg/kg/day for 10 days, followed by tapering over next 4 days. None of the patients received albendazole therapy. The patients were followed up monthly, at least for 9 months. A repeat CT scan was performed after 6 months. The data were analysed by chi-square test. The majority of patients were young. Simple partial seizure, with or without secondary generalization, was the commonest seizure type encountered. Follow-up CT scans at 6 months demonstrated non-significantly better response for prednisolone treated patients. In prednisolone group the lesion disappeared in 52% of patients and in 48% patients who received placebo. However, a significantly lesser number of prednisolone treated patients (n=12%) than controls (n=48%), had seizure recurrence. Our study suggests that short-term prednisolone therapy may not help in rapid resolution of solitary cysticercus granuloma, however, prednisolone therapy improves seizure-related prognosis. PMID- 16269180 TI - Rapid detection of pneumococcal antigen in serum samples for diagnosing pneumococcal pneumonia. AB - OBJECTIVES: The aim of the study is to assess the usefulness of C polysaccharide and polysaccharide capsular antigen detection by immunochromatography (ICT) and enzyme immunoassay (EIA), respectively, in serum samples for diagnosing pneumococcal pneumonia. METHODS: Adult patients included in the study were classified in the following groups: In group 1 we studied 101 serum samples from patients with pneumonia due to Streptococcus pneumoniae. In 53 cases the pneumonia was bacteremic. The second group contained 113 serum samples from patients with no pneumococcal pneumonia. Group 3 was made up of 40 serum samples from healthy subjects with no clinical or radiological signs of pneumonia. RESULTS: Using ICT, antigen was detected in 50% of patients with pneumococcal pneumonia, in 64.3 and 40.9% of patients with bacteremic and non-bacteremic pneumococcal pneumonia, respectively. Using EIA, antigens were detected in 35.8% of patients with pneumococcal pneumonia, in 45 and 22.2% of patients with bacteremic and non-bacteremic pneumococcal pneumonia, respectively. CONCLUSIONS: In conclusion, the sensitivity of the tests is low. However, in special situations, where obtaining large volume of urine is difficult, they could be a complementary method in the rapid diagnosis of pneumococcal pneumonia. PMID- 16269181 TI - Brucellosis is not a major cause of febrile illness in patients at public health care facilities in Binh Thuan Province, Vietnam. AB - OBJECTIVE: To determine the presence of brucellosis among patients with acute febrile illness at health care facilities in Binh Thuan province, Vietnam. METHOD: A retrospective seroepidemiological study on serum samples collected at 13 not adjacent health care facilities using the Rose Bengal test as a rapid screening test and the Brucella IgM/IgG flow assay as a simple confirmatory test. RESULT: The seroprevalence in the Rose Bengal test among 406 patients presented with acute undifferentiated fever was 14.8%. Seven of the 64 Rose Bengal test positive samples reacted weakly (1+) positive in the Brucella IgM/IgG flow assay. No seroconversion was observed. CONCLUSIONS: Brucellosis is not a major cause of morbidity in Binh Thuan province. PMID- 16269182 TI - Acute fulminant hepatitis B in a patient with diabetic nephropathy treated successfully with concomitant lamivudine and molecular adsorbents recirculating system. AB - A 36-year-old man with type 2 diabetes and diabetic nephropathy treated with hemodialysis developed hepatitis B virus (HBV)-induced acute fulminant hepatic failure (FHF). Despite supportive treatment, the condition rapidly progressed as manifested by severe jaundice, coagulopathy and hepatic coma. He was placed on the waiting list for liver transplantation and was treated with lamivudine and extracoporeal liver support with the molecular adsorbent recirculating system (MARS). After three 8-h sessions of MARS treatment in 1 week, he had remarkable improvement in clinical symptoms and serum biochemistry. On the 14th hospital day, surface antigen seroconversion was noted with undetectable hepatitis B virus surface antigen (HBs Ag) and low titre of anti-HBs antibody, indicating a complete recovery from acute fulminant hepatitis B. MARS treatment has been reported to benefit patients with liver failure from different causes including acute exacerbation of chronic hepatitis B, poisoning, post transplantation and Wilson's disease. The present case suggests its potential benefit when combined with lamivudine in treating uremic patients with acute fulminant hepatitis B. PMID- 16269183 TI - Is malaria a predisposing factor for third molar pericoronitis in the tropics? AB - BACKGROUND: Malaria is an important disease in the tropics, and its role as a predisposing factor or co morbidity has been investigated in many diseases including HIV infection and tuberculosis. There are very few studies, which have investigated its role in oral and dental diseases. Our study aimed to demonstrate the possible role of malaria in predisposing to pericoronitis, an infection affecting impacted third molars predominantly. PATIENTS AND METHODS: Thirty-eight patients presenting with pericoronitis were tested for malaria parasites and results compared with that obtained from controls that were equally susceptible to pericoronitis but did not have the infection. RESULTS: 19.7% of the study group compared to 6.6% of control group had malaria parasite in their blood. This difference was statistically significant, P=0.018 (Fisher's exact). The odds ratio was 4.3 (95% CI=1.2-17.0). CONCLUSIONS: Malaria appears to be a predisposing factor to pericoronitis in this study. There is a need for further studies on the possible role of malaria in oral and dental diseases. PMID- 16269184 TI - Pegylated interferon alpha-2b plus ribavirin for the treatment of chronic hepatitis C in HIV-coinfected patients. AB - OBJECTIVES: HIV-coinfection accelerates the course of HCV-related liver disease. Since, highly active anti-retroviral therapy significantly improved survival of HIV-patients more coinfected patients develop end stage liver disease. Therefore, treatment options for chronic hepatitis C in HIV-coinfected patients need to be evaluated. METHODS: Efficacy and safety of pegylated interferon alpha-2b (peg IFN) plus ribavirin (RBV) was examined within this prospective, uncontrolled, multicentre trial. Patients received peg IFN (1.5 microg/kg) once weekly plus RBV 800 mg daily for 48 weeks for HCV genotypes (GT) 1/4 and 24 weeks for GT 2/3. RESULTS: One hundred and twenty-two patients were enrolled. Patients were predominantly male (68%) and former i.v. drug users (61%). Baseline characteristics (median) were as follows: age 39 years (range 23-58), CD4 count 494 cells/microl (range 150-1578/microl), HIV-RNA 2.3log copies/ml (range <1.7 5.4log copies/ml). 61% currently received anti-retroviral treatment. Fifty-six percent had HCV GT 1. EOT response was achieved by 52%. However, only 25% achieved sustained response (SR) due to a high relapse rate. SR rates were significantly higher among patients with GT 2/3 compared to those with GT 1/4 (44 vs. 18%). SR was observed in only one patient without early response (ER). Discontinuation rate was 30%, 21% discontinued due to adverse events. CONCLUSION: Peg IFN/RBV appears safe and effective in HIV/HCV-coinfected patients. GT 2/3 is associated with better SR. Lack of ER strongly predicts non-response. High relapse rates substantially reduce treatment success. In terms of toxicity neuro psychiatric side effects frequently required treatment discontinuation. PMID- 16269186 TI - The metabolism of structured triacylglycerols. AB - The triacylglycerol (TAG) structure in addition to the overall fatty acid profile is of importance when considering the nutritional effect of a dietary fat. This review aims at summarizing our current knowledge of the digestion, absorption, uptake, and transport of structured TAGs, with particular emphasis on the following aspects: gastric emptying, specificity of pancreatic lipase, lymphatic transport and clearance of chylomicrons, effects of lipid structure on tissue lipid compositions and the fecal loss of fats. So an overview will be provided for how the structure and fatty acid composition of TAGs affect their absorption and the distribution of the fatty acids in the body following digestion and absorption. PMID- 16269185 TI - Visceral leishmaniasis (kala-azar)--the Bihar (India) perspective. AB - From a hospital-based surveillance carried out in Rajendra Memorial Research Institute of Medical Sciences, Patna, Bihar, India, the socio-economic, demographic and treatment response information of 737 patients admitted with visceral leishmaniasis (VL) during January 2001-December 2003, were analysed. The disease was two times higher in males than in females because of several factors including clothing pattern, sleeping habits and occupation. In Bihar, the second poorest state in India, poverty plays a major role in perpetuation of the disease, contributing to malnutrition, illiteracy (60%), and poor housing (82%). Further, presences of peri-domestic animal shelters around houses (63%) and vegetations (77%) facilitate breeding of sand fly vector. Clinical and laboratory characteristics were similar in the age groups <12 years and >12 years. The increasing unresponsiveness of VL patients to conventional anti-leishmanial drugs, e.g. sodium antimony gluconate (SAG) and pentamidine, has definitely posed a major therapeutic challenge in combating the disease. Amphotericin B, though costly, is highly effective. Miltefosine is a highly promising new oral drug for VL. PMID- 16269187 TI - Immunohistochemical visualization of neurons and specific glial cells for stereological application in the porcine neocortex. AB - The pig is becoming an increasingly used non-primate model in basic experimental studies of human neurological diseases. In spite of the widespread use of immunohistochemistry and cell type specific markers, the application of immunohistochemistry in the pig brain has not been systematically described. Therefore, to facilitate future stereological studies of the neuronal and glial cell populations in experimental neurological diseases in the pig, we established a battery of immunohistochemical protocols for staining of perfusion fixed porcine brain tissue processed as free floating cryostat-, vibratome- or paraffin sections. Antibodies against NeuN, GFAP, S100-protein, MBP, CNPase, CD11b, CD68 (KP1), CD45 and Ki67 were evaluated, and all except CD68 and CD45 resulted in staining of high quality in either type of tissue. Each staining was evaluated with respect to specificity and sensitivity in identification of the individual cells, and for penetration of the staining and maintenance of section thickness above 25 microm, necessary for stereological cell counting. In the cases of NeuN, CNPase, CD11b and Ki67 the staining met the demands to be applicable in stereological analyses using the optical disector. In conclusion, all protocols will be applicable in studies of pathological and neurochemical changes in the porcine brain, and a few protocols applicable for stereology. PMID- 16269188 TI - Testing for directed influences among neural signals using partial directed coherence. AB - One major challenge in neuroscience is the identification of interrelations between signals reflecting neural activity. When applying multivariate time series analysis techniques to neural signals, detection of directed relationships, which can be described in terms of Granger-causality, is of particular interest. Partial directed coherence has been introduced for a frequency domain analysis of linear Granger-causality based on modeling the underlying dynamics by vector autoregressive processes. We discuss the statistical properties of estimates for partial directed coherence and propose a significance level for testing for nonzero partial directed coherence at a given frequency. The performance of this test is illustrated by means of linear and non linear model systems and in an application to electroencephalography and electromyography data recorded from a patient suffering from essential tremor. PMID- 16269190 TI - Physiological relevance of constitutive activity of 5-HT2A and 5-HT2C receptors. AB - It is generally accepted that seven-transmembrane receptors have the capacity to regulate cellular signaling systems in the absence of occupancy by a ligand (i.e. the receptors display constitutive activity). Drugs can increase (agonists), decrease (inverse agonists) or not change (antagonists) receptor activity towards a cellular effector. Moreover, some drugs (protean ligands) have multiple pharmacological properties (e.g. agonism towards one response and inverse agonism towards another response coupled to the same receptor and measured from the same cells, simultaneously). In this article, we describe response-dependent constitutive activity and ligand pharmacology for 5-HT2A and 5-HT2C receptors in vitro. Moreover, we provide evidence that 5-HT2A and 5-HT2C receptor constitutive activity is physiologically relevant in vivo and suggest that strong consideration should be given to the impact of constitutive receptor activity on disease and the therapeutic potential of inverse agonism. PMID- 16269189 TI - Pivotal role of endogenous tachykinins and the NK1 receptor in mediating leukocyte accumulation, in the absence of oedema formation, in response to TNFalpha in the cutaneous microvasculature. AB - Tachykinins including substance P (SP) are well known to play a role in influencing oedema formation and leukocyte accumulation during tissue insult and inflammation. Cutaneous inflammatory models to characterize a TNFalpha-dependent mechanism where endogenous SP act via the NK1 receptor to promote leukocyte accumulation in the absence of oedema formation were used. We found that TNFalpha induced dose-dependent leukocyte accumulation at 4 h, which returned towards basal levels at 8 h in NK1+/+ mice. This response was absent in both the NK1+/+ mice treated with an NK1 receptor antagonist and NK1-/- mice. At the highest dose IL-6 induced a significant accumulation in NK1+/+ and NK1-/- mice but IL-12 was ineffective. SP induced skin oedema but none of the cytokines did. Either co injection of SP with low dose of TNFalpha (0.3 pmol/site) or SP previously injected (30 min) to TNFalpha evoked a significant increase in MPO activity when compared with that induced by the cytokine alone. In contrast, SP injected i.d. 3.5 h after TNFalpha failed to produce additive response. Control, but not capsaicin-pretreated rats (to deplete sensory nerves), exhibited a marked increase in MPO activity in response to TNFalpha. Histological analysis showed that TNFalpha caused tissue infiltrate of leukocytes in NK1+/+ mice, whilst leukocytes accumulated at intravascular sites in NK1-/- mice, but did not appear to emigrate, suggesting a defect in trans-endothelial migration. Interestingly, monocytes in addition to neutrophils accumulated 4 h post TNFalpha injection. In conclusion, the NK1 receptor plays a functional role in mediating leukocyte accumulation independently of the historically important NK1 mediated oedema formation. It seems that TNFalpha directly activates sensory nerve in addition to its chemoattractant activity. The NK1 receptor agonist influences the accumulation of monocytes in addition to that of PMN by 4 h, thus revealing an important influence of the NK1 receptor on TNFalpha mediated events in mouse skin. PMID- 16269191 TI - A vector series for rapid PCR-mediated C-terminal in situ tagging of Trypanosoma brucei genes. PMID- 16269192 TI - Public health and policy. AB - Antimicrobial agent usage data are essential for focusing efforts to reduce misuse and overuse of antimicrobial agents in food producing animals because these practices may select for resistance in bacteria of animals. Transfer of resistant bacteria from animals to humans can lead to human infection caused by resistant pathogens. Resistant infections can lead to treatment failures, resulting in prolonged or more severe illness. Multiple World Health Organization (WHO) reports have concluded that both antimicrobial resistance and antimicrobial usage should be monitored on the national level. The system for collecting antimicrobial usage data should be clear and transparent to facilitate trend analysis and comparison within and among countries. Therapeutic, prophylactic and growth promotion use should be recorded, along with route of administration and animal species and/or production class treated. The usage data should be compared to resistance data, and the comparison should be made available in a timely manner. In the United States, surveillance of antimicrobial resistance in foodborne bacteria is performed by the National Antimicrobial Resistance Monitoring System (NARMS) for enteric bacteria, however, the United States still lacks a mechanism for collecting antimicrobial usage data. Combined with antimicrobial resistance information from NARMS, antimicrobial usage data will help to direct education efforts and policy decisions, minimizing the risk that people will develop antimicrobial resistant infections as a result of eating food of animal origin. Ultimately mitigation strategies guided by usage data will be more effective in maintaining antimicrobial drugs for appropriate veterinary use and in protecting human health. PMID- 16269193 TI - Fluorescent proteins as a toolkit for in vivo imaging. AB - Green fluorescent protein (GFP) from the jellyfish Aequorea victoria, and its mutant variants, are the only fully genetically encoded fluorescent probes available and they have proved to be excellent tools for labeling living specimens. Since 1999, numerous GFP homologues have been discovered in Anthozoa, Hydrozoa and Copepoda species, demonstrating the broad evolutionary and spectral diversity of this protein family. Mutagenic studies gave rise to diversified and optimized variants of fluorescent proteins, which have never been encountered in nature. This article gives an overview of the GFP-like proteins developed to date and their most common applications to study living specimens using fluorescence microscopy. PMID- 16269194 TI - Novel extraction strategy of ribosomal RNA and genomic DNA from cheese for PCR based investigations. AB - Cheese microorganisms, such as bacteria and fungi, constitute a complex ecosystem that plays a central role in cheeses ripening. The molecular study of cheese microbial diversity and activity is essential but the extraction of high quality nucleic acid may be problematic: the cheese samples are characterised by a strong buffering capacity which negatively influenced the yield of the extracted rRNA. The objective of this study is to develop an effective method for the direct and simultaneous isolation of yeast and bacterial ribosomal RNA and genomic DNA from the same cheese samples. DNA isolation was based on a protocol used for nucleic acids isolation from anaerobic digestor, without preliminary washing step with the combined use of the action of chaotropic agent (acid guanidinium thiocyanate), detergents (SDS, N-lauroylsarcosine), chelating agent (EDTA) and a mechanical method (bead beating system). The DNA purification was carried out by two washing steps of phenol-chloroform. RNA was isolated successfully after the second acid extraction step by recovering it from the phenolic phase of the first acid extraction. The novel method yielded pure preparation of undegraded RNA accessible for reverse transcription-PCR. The extraction protocol of genomic DNA and rRNA was applicable to complex ecosystem of different cheese matrices. PMID- 16269195 TI - Antifungal activity of TiO2 photocatalysis against Penicillium expansum in vitro and in fruit tests. AB - The antifungal activity of TiO2 photocatalytic reaction in the form of TiO2 powder and TiO2 coated on a plastic film against Penicillium expansum was investigated in vitro and in fruit tests. The mixture of P. expansum conidial suspension and TiO2 powder was added to potato dextrose agar (PDA) plates for vitro test. The TiO2 photocatalytic reaction reduced conidial germination of the fungal pathogen. It was found that the ability of the TiO2 photocatalytic reaction to suppress P. expansum growth correlated to the amount of TiO2 added. Lower numbers of viable colonies of P. expansum were observed with increasing amount of TiO2. Regardless of the kind of selected fruit inoculated with P. expansum, both TiO2 powder and TiO2-coated film exhibited antifungal activity to control fruit rot. Development of Penicillium rot in apple was significantly (P = 0.05) retarded by the TiO2 photocatalytic reaction. Similarly the TiO2 photocatalytic reaction was the only treatment where no tomato fruit rot was noticeable after 1 week of storage. TiO2-coated film also decreased brown lesions and Penicillium rot infection in lemons. The mean severity fruit rot scores (browning and softening flesh) were 3.2 and 1.9 for uncoated and TiO2-coated film, respectively. Our findings suggest that "TiO2 photocatalytic reaction" shows antifungal activity against P. expansum which may have potential for postharvest disease control. PMID- 16269196 TI - Isolation and characterization of verocytotoxin-producing Escherichia coli O157 from Turkish cattle. AB - The objective of this study was to collect rectal swabs from the cattle in a slaughterhouse located in Hatay (Turkey) immediately after slaughter for the isolation and characterization of verotoxin-producing Escherichia coli O157 in each month during a 1-year period. The rectal swab samples were analyzed for the isolation of E. coli O157 through pre-enrichment, immunomagnetic separation and selective plating on CT-SMAC agar. E. coli O157 was isolated from 77 (13.6%) of the samples. The presence of E. coli O157 changed during a 1-year period, in that the occurrence of E. coli O157 was the highest in July and November and lowest in February. A total of 66 isolates out of 77 were serotype O157:H7 and 11 were serotype O157:NM. PCR analysis of E. coli O157 virulence genes revealed that all O157:H7/NM were positive for rbf(O157), 74 positive for EhlyA, 72 positive for eaeA, 62 positive for vtx2, and 3 positive for both vtx1 and vtx2. It was presented by cytotoxicity tests that many of E. coli O157 isolates showed high cytotoxicity on Vero cells. All of the isolates containing EhlyA showed enterohaemolysin production. PMID- 16269197 TI - Evolution of ORF5 of Spanish porcine reproductive and respiratory syndrome virus strains from 1991 to 2005. AB - ORF5 sequences of porcine reproductive and respiratory syndrome virus (PRRSV) were analysed to determine genetic diversity, codon usage, positive and negative selection sites and potential changes in the predicted glycoprotein 5 (GP5). A hypothetical GP5 containing all selected sites was constructed to determine its characteristics. These sequences corresponded to isolates obtained 10 years apart (1991-1995, 18 strains) and a second set (n = 46) from 2000 to 2005. Similarity to Lelystad virus (LV) decreased from 95.5% in 1991-1995 to 89.5% in 2000-2005. Three highly variable regions were found in ORF5. Codon usage was different in both sets for leucine, glutamine, serine and proline. Thus, 2000-2005 sequences used codons more similar to those present in highly expressed pig genes compared to the 1991-1995 set. Twenty four sites of positive selection and 20 sites of negative selection were found in GP5, most of them in transmembrane regions. Additional glycosylation in N37 of GP5 was common in 2000-2005 but some sequences lack a glycosylation site in N46. The hypothetical GP5 was only 88.1% similar to LV and was less hydrophobic. Taking together these results suggest that PRRSV is still adapting to pig cells. PMID- 16269198 TI - Chemotherapy based on HPMA copolymer conjugates with pH-controlled release of doxorubicin triggers anti-tumor immunity. AB - A novel class of anti-cancer therapeutics - polymeric conjugates of N-(2 hydroxypropyl)methacrylamide (HPMA) copolymers and doxorubicin with pH-controlled release of the drug - is highly efficient in killing tumor cells in vitro and is potent in eradicating growing tumors in vivo. Moreover, in comparison with low molecular-weight drugs, the macromolecular therapeutics show decreased acute as well as delayed adverse side-toxicity. More importantly, the polymeric conjugates trigger the onset of specific anti-tumor immune response and this anti-tumor immunity can be transferred with splenocytes to naive recipients. In other terms, chemotherapy based on conjugates of HPMA copolymer with doxorubicin possesses immunomodulating properties. This finding might also have wider implications for the management of relapsing tumors in human patients. PMID- 16269199 TI - Enhancement of bronchial octreotide absorption by chitosan and N-trimethyl chitosan shows linear in vitro/in vivo correlation. AB - Chitosan is a biocompatible polysaccharide of natural origin that can act as a permeation enhancer. In this study, we used an integral in vitro/in vivo correlation approach to: a) investigate polysaccharide-mediated absorption kinetics of the peptide drug octreotide across mammalian airway epithelium, b) assess formulation toxicity, c) correlate the mechanism of permeation enhancement. The 20% and 60% N-trimethylated chitosan derivatives (TMC20 and TMC60) were synthesized by alkaline methylation using chitosan as starting material. Octreotide was administered in control buffers or in 1.5% (w/v) gel phase formulations of pH 5.5 for chitosan and pH 7.4 for TMCs. In vitro, reconstituted Calu-3 cell monolayers were used for trans-epithelial electrical resistance (TEER), transport and cytotoxicity assays. Intratracheal instillation in rats was used to determine octreotide kinetics and formulation toxicity in vivo. Chitosan, TMC20 and TMC60 decreased TEER significantly and enhanced octreotide permeation in vitro by 21-, 16- and 30-fold. In vivo, sustained release properties of the formulations were observed and the bio-availability was enhanced by 2.4-, 2.5- and 3.9-fold, respectively. Interestingly, we found a linear in vitro/in vivo correlation between calculated absorption rates (R2=0.93), suggesting that the permeation enhancement by polysaccharides, both in vitro and in vivo, proceeds via an analogous mechanism. Cell viability and histology studies showed that the TMCs are safer than chitosan and that Calu-3 cell monolayers are a valuable model for predicting paracellular transport kinetics in airway epithelia. Additionally, cationic polysaccharides are promising enhancers for peptide drug absorption with prospect for sustained release formulations. PMID- 16269200 TI - Biodegradable microspheres alone do not stimulate murine macrophages in vitro, but prolong antigen presentation by macrophages in vitro and stimulate a solid immune response in mice. AB - The purpose of this study was to analyze the potential of various types of biodegradable microspheres (MS) (i) to activate in vitro cell line-derived macrophages (RAW 264.7, Mphi), and primary peritoneal and bone marrow-derived mouse Mphi, to prolong the release and presentation of microencapsulated synthetic malaria antigens by Mphi after uptake of antigen-loaded MS, and (ii) to stimulate an immune response in mice against a microencapsulated synthetic malaria antigen. The MS were made of various types of poly(lactide-co-glycolide) (PLGA) or chitosan cross-linked with tripolyphosphate. PLGA, but not chitosan MS, were efficiently ingested by Mphi. Upon exposure to the various MS types, Mphi increased only the production of reactive oxygen intermediates (ROI), while the production of nitric oxide (NO), tumor necrosis factor alpha (TNF-alpha), and the expression of cyclooxigenase-2 (COX-2), inducible NO synthase (iNOS), the cell surface markers MHC class I and II, and CD 86 remained unaffected. In vitro release of the microencapsulated antigen from PLGA50:50 MS followed a pulsatile pattern and extended over 14 weeks. This prolonged antigen release was also mirrored in the significantly prolonged antigen presentation over more than 7 days by Mphi after uptake of antigen-loaded PLGA MS. Finally, antigen-loaded PLGA MS induced a solid immune response in mice after a single s.c.-injection, which was only slightly inferior to the antibody titers measured with the control formulation with Montanide ISA720. These results suggest that MS are well tolerated by Mphi. The prolonged antigen presentation by Mphi, as measured in vitro, along with the capacity to induce a strong immune response in animals emphasize that biodegradable MS are a very promising delivery system for both preventive and immunotherapeutic vaccines. PMID- 16269201 TI - Synthesis and application of a non-viral gene delivery system for immunogene therapy of cancer. AB - The synthesis and gene delivery application of a novel lipopolymer, PEG-PEI-CHOL (PPC), is described. PPC is composed of a low molecular weight branched polyethylenimine (PEI) covalently linked with functional groups methoxypolyethyleneglycol (PEG) and cholesterol (CHOL). The potential utility of PPC as a gene delivery polymer was evaluated by showing its ability to form stable nanocomplexes with DNA, protect DNA from degradation by DNase and mediate gene transfer in vitro and in vivo in solid tumors. The ratio of PEG/PEI/CHOL and nitrogen to phosphate (Polymer/DNA) was optimized for physico-chemical properties and gene delivery efficiency of PPC/DNA complexes. The gene therapy application of the polymer was shown following administration of a murine IL-12 plasmid (pmIL 12) formulated with PPC into tumors in mice which resulted in significant inhibition of tumor growth. The inhibitory effects of pmIL-12/PPC were enhanced when combined with specific chemotherapeutic agents, demonstrating the potential usefulness of pIL-12/PPC as an adjuvant therapy for cancer treatment. PMID- 16269202 TI - An evolutionary scenario for one of the largest yeast gene families. AB - The DUP gene family of Saccharomyces cerevisiae comprises 23 members that can be divided into two subfamilies--DUP240 and DUP380. The location of the DUP loci suggests that at least three mechanisms were responsible for their genomic dispersion: nonreciprocal translocation at chromosomal ends, tandem duplication and Ty-associated duplication. The data we present here suggest that these nonessential genes encode proteins that facilitate membrane trafficking processes. Dup240 proteins have three conserved domains (C1, C2 and C3) and two predicted transmembrane segments (H1 and H2). A direct repetition of the C1-H1-H2 C2 module is observed in Dup380p sequences. In this article, we propose an evolutionary model to account for the emergence of the two gene subfamilies. PMID- 16269203 TI - Different expression role among glutamate transporters in rat retinal glial cells under various culture conditions. AB - Using cultured rat retinal glial cells, the changes in the expression of glutamate transporters (GLTs) under such conditions as the degree of confluence of the cells, hypoxia, glutamate loading, and potassium loading, were assessed. After the retinal glial cells were isolated from 3-day-old Sprague-Dawley rats, GLAST, GLT-1, and EAAC1 mRNA expression was confirmed by reverse transcription polymerase chain reaction (RT-PCR) and their protein expression was confirmed by Western blot analysis and immunostaining. Changes in the expression of the GLTs at 6 h after passage and at confluence, during culture in 5% oxygen, during glutamate loading and during potassium loading were assessed using real-time PCR. Although the GLAST mRNA expression was increased during glutamate and potassium loading, no changes in the expression were observed during hypoxia and at confluence. By contrast, the GLT-1 mRNA expression was increased during hypoxia and at confluence, but not during glutamate and potassium loading, and the EAAC1 mRNA expression was not changed only during glutamate loading. The expression of EAAC1 in the cultured retinal glial cells was confirmed. The expression of the GLTs varied according to the environment and the type of load suggests that the involvement of the GLTs in retinal physiology and pathology varies depending on the subtype. PMID- 16269204 TI - Role of melanocortin in the long-term regulation of energy balance: lessons from a seasonal model. AB - Siberian hamsters express photoperiod-regulated seasonal cycles of body weight and food intake, providing an opportunity to study the role of melanocortin systems in regulating long-term adaptive changes in energy metabolism. These hamsters accumulate intraperitoneal fat reserves when kept in long summer photoperiods, but show a profound long-term decrease in food intake and body weight when exposed to a short winter photoperiod. Icv administration of a MC3/4 R agonist (MTII) potently suppresses food intake in hamsters in both the obese and lean state, indicating the potential for melanocortin systems to regulate energy metabolism in the hypothalamus of the Siberian hamster. Icv treatment with the melanocortin antagonist SHU9119 increases food intake in both seasonal states. Moreover, hamsters bearing neurotoxic lesions, which destroy the majority of POMC expressing neurons in the arcuate nucleus are still able to show seasonal regulation of body weight. These studies in a seasonal model substantiate the view that endogenous melanocortin systems exert a tonic inhibition of food intake in mammals. The observations that this melanocortin tone occurs to a similar extent in both an anabolic state induced by a long day photoperiod, and in a catabolic state induced by a short day photoperiod, suggests that alterations in endogenous melanocortin tone are not the primary cause of the lipolysis, weight loss and hypophagia which characterize the establishment of the short day-induced overwintering state. PMID- 16269205 TI - Characterisation of CTL and IFN-gamma synthesis in ponies following vaccination with a NYVAC-based construct coding for EHV-1 immediate early gene, followed by challenge infection. AB - Equine herpesvirus-1 (EHV-1) is a ubiquitous pathogen of horses, which continues to cause respiratory and neurological disease and abortion, despite the widespread use of vaccines. Cell mediated immunity (CMI) is thought to play a major role in protection against infection with EHV-1. The aim of this study was to characterise the virus-specific CMI response in ponies vaccinated with vP1014, a vaccinia-based construct (NYVAC) coding for the immediate early gene (gene 64) of EHV-1. This gene product is a CTL target protein for an equine MHC class I allele expressed on the A3 haplotype. EHV-primed yearling ponies expressing this haplotype were vaccinated once (n = 1), three (n = 1), or four times (n = 2), and one pony was kept as an unvaccinated control. Cytotoxic T lymphocyte (CTL) activity and interferon gamma (IFN-gamma) synthesis were measured before and after vaccination and challenge infection with EHV-1. Multiple immunisations with vP1014 resulted in increased CTL activity and IFN-gamma synthesis specific for EHV-1 compared with unvaccinated or singly vaccinated ponies. The phenotype of EHV-1 specific T-cells synthesising IFN-gamma was also modified by immunisation. In the unvaccinated pony, the predominant population synthesising IFN-gamma after EHV-1 stimulation was CD8alpha+. In contrast, multiply vaccinated ponies demonstrated an increased proportion of CD8alpha- T-cells synthesising IFN-gamma. The results demonstrated that vaccination with a NYVAC-based construct coding for gene 64 stimulated CMI. This immune response alone did not protect against challenge infection. However, the study does illustrate that vaccinia-based vaccines can stimulate CMI in the horse and may therefore contribute to protection against disease caused by EHV-1. PMID- 16269206 TI - Preparation and characterization of SARS in-house reference antiserum. AB - A reference antiserum for SARS is in urgent need in the development of SARS vaccine and other serological test of SARS research. Convalescent serum was collected from clinical confirmed patient. ELISA, Western-blotting and neutralization assay detected specific antibody against SARS coronavirus. This antiserum was prepared as in-house reference antiserum, freeze-dried and sealed in ampoules. The potency of this reference antiserum is defined to be 52.7 U after extensive calibration. Further, collaborative studies for the evaluation of this serum are needed in order to satisfy the requirements for international reference antiserum. PMID- 16269207 TI - Changes in foot and lower limb coupling due to systematic variations in step width. AB - BACKGROUND: Motion at the midfoot joints can contribute significantly to overall foot motion during gait. However, there is little information regarding the kinematic coupling relationship at the midfoot. The purpose of the present study was to determine whether the coupling relationship at the midfoot and subtalar joints was affected when step width was manipulated during running. METHODS: Twelve subjects ran over-ground at self-selected speeds using three different step widths (normal, wide, cross-over). Coupling at the midfoot (forefoot relative to rearfoot) and subtalar (rearfoot relative to shank) joints was assessed using cross-correlation techniques. FINDINGS: Rearfoot kinematics were significantly different from normal running in cross-over running (P<0.05) but not in wide running. However, coupling between rearfoot eversion/inversion and shank rotation was consistently high (r>0.917), regardless of step width. This was also the case for coupling between rearfoot frontal plane motion and forefoot sagittal plane (r<-0.852) and forefoot transverse plane (r>0.946) motion. There was little evidence of coupling between rearfoot frontal plane motion and forefoot frontal plane motion in any of the conditions. INTERPRETATION: Forefoot frontal plane motion appeared to have little effect on rearfoot frontal plane motion and thus, had no effect on motion at the subtalar joint. The strong coupling of forefoot sagittal and transverse plane motions with rearfoot frontal plane motion suggests that forefoot motion exerts an important influence on subtalar joint kinematics. PMID- 16269208 TI - Ankle function during gait in patients with chronic ankle instability compared to controls. AB - BACKGROUND: Despite much research, the reasons behind the development of chronic ankle instability in individuals post ankle inversion sprain are unknown. Chronic ankle instability has not previously been investigated dynamically using 3D motion analysis during walking. We hypothesised that chronic ankle instability subjects would exhibit a different kinematic and kinetic pattern during normal walking when compared with a control group. METHODS: Gait analysis was carried out on fifty subjects (25 chronic ankle instability, and 25 age, gender, activity, and gait velocity matched controls) during walking. Kinematic and kinetic pattern differences using the 3D motion analysis system combined with a force plate were established during 100 ms pre-heel strike and 200 ms post-heel strike, between the chronic ankle instability subjects and controls during normal walking. FINDINGS: Chronic ankle instability subjects were significantly (P<0.01) more inverted in the frontal plane compared to controls from 100 ms pre-heel strike to 200 ms post-heel strike. The joint angular velocity was significantly (P<0.05) higher at heel strike in the chronic ankle instability group. During the early stance phase of gait chronic ankle instability subjects appear to be controlled by an evertor muscle moment working concentrically compared to an invertor muscle moment working eccentrically in the controls. INTERPRETATION: These changes in kinematics and kinetics which arise are likely to result in increased stress being applied to ankle joint structures during the heel strike and loading response phases of the gait cycle. This could result in repeated injury and consequent damage to ankle joint structures. PMID- 16269209 TI - Role of amino acid properties to determine backbone tau(N-Calpha-C') stretching angle in peptides and proteins. AB - The analysis of the basic geometry of amino acid residues of protein structures has demonstrated the invariability of all the bond lengths and bond angles except for tau, the backbone N-Calpha-C' angle. This angle can be widened or contracted significantly from the tetrahedral geometry to accommodate various other strains in the structure. In order to accurately determine the cause for this deviation, a survey is made for the tau angles using the peptide structures and the ultrahigh resolution protein structures. The average deviation of N-Calpha-C' angles from tetrahedral geometry for each amino acid in all the categories were calculated and then correlated with forty-eight physiochemical, energetic and conformational properties of amino acids. Linear and multiple regression analysis were carried out between the amino acid deviation and the 48 properties. This study confirms the deviation of tau angles in both the peptide and protein structures but similar forces do not influence them. The peptide structures are influenced by physical properties whereas as expected the conformational properties influence the protein structures. And it is not any single property that dominates the deviation but the combination of different factors contributes to the tau angle deviation. PMID- 16269210 TI - Biphasic effects of exogenous VEGF on VEGF expression of adult neural progenitors. AB - Vascular endothelial growth factor (VEGF) regulates neurogenesis. The present study investigated the direct effect of VEGF on the enhancement of proliferation and differentiation of the adult mouse subventricular zone (SVZ) neural progenitors in vitro. A high dose (500 ng/ml) of VEGF significantly downregulated endogenous VEGF receptors 1 and 2, which was associated with significantly reduced neural progenitor cell proliferation and enhancement of neuronal differentiation. A low dose (50 ng/ml) of VEGF significantly upregulated endogenous VEGF receptors 1 and 2 but did not increase proliferation and differentiation. These data suggest that exogenous VEGF has a biphasic effect on the expression of endogenous VEGF receptors, and the high dose of VEGF enhances adult neural progenitor cell differentiation into neurons. PMID- 16269211 TI - GABA(A) receptor gamma subunits in the hippocampus of the rat after perforant pathway lesion. AB - Immunohistochemical and Western blotting techniques were employed to examine the alterations in immunostaining of the gamma-amino butyric acid (GABA) receptor subunits gamma 1/3 and 2 within the hippocampus of the rat brain at 1, 3, 7, 14, and 30 days after a unilateral perforant pathway lesion. At 1, 3, and 7 days post lesion, we observed a remarkable decrease in gamma 1/3 neuropil staining in the deafferented zone (i.e., the outer molecular layer of the dentate gyrus ipsilateral to the lesion), although at 3 and 7 days post-lesion, staining intensity was considerably recovered. At 14 days post-lesion, the gamma 1/3 immunostaining was indistinguishable from that of controls and it appeared yet more robust at 30 days post-lesion. We also observed a slight decrease in gamma 2 neuropil staining until 7 days post-lesion, and an increase in gamma 2 staining at 30 days post-lesion. Western blot analysis demonstrated data that was relatively consistent with our immunohistochemical observations, although gamma 3 was hardly detectable. Our study suggests that gamma subunits of the GABA(A) receptor in the dentate gyrus display a plastic response to the deafferentation of the perforant pathway. PMID- 16269212 TI - Identification of distinct characteristics of postural sway in Parkinson's disease: a feature selection procedure based on principal component analysis. AB - We selected descriptive measures of the centre of pressure (CoP) displacement in quiet standing, by means of a procedure based on principal component analysis, in two groups particularly different in terms of postural behaviours, such as subjects with Parkinson's disease (PD) in the levodopa off and on states. We computed 14 measures of the CoP: 5 measures of CoP trajectory over the support surface, 3 measures that estimated the area covered by the CoP, 1 measure that estimated the principal CoP sway direction, 1 measure that quantified the CoP total power, 1 measure that estimated the variability of CoP frequency content and 3 measures of characteristic CoP frequencies [L. Rocchi, L. Chiari, A. Cappello, Feature selection of stabilometric parameters based on principal component analysis, Med. Biol. Eng. Comput. 42 (2004) 71-79; L. Rocchi, L. Chiari, F.B. Horak, Effects of deep brain stimulation and levodopa on postural sway in Parkinson's disease, J. Neurol. Neurosurg. Psychiatry, 73 (2002) 267 274]. The feature selection, independently applied to the measures obtained in the two groups, resulted in different principal component (PC) subspaces of the 14-dimension original data set (4 PCs in the off and 3 PCs in the on state to account for over 90% of the original variance), but in the same 5 CoP measures (selected features) needed to describe the different postural behaviours: root mean square distance; mean velocity; principal sway direction; centroidal frequency of the power spectrum; frequency dispersion. The five selected features were found to provide insight into the postural control mechanisms and to describe changes in postural strategies in the two groups of PD subjects, off and on levodopa. Thus, the five selected features may be recommended for use in clinical practice and in research, in the direction toward the definition of a standard protocol in quantitative posturography. PMID- 16269213 TI - Nicotine prevents HIVgp120-caused electrophysiological and motor disturbances in rats. AB - Human immunodeficiency virus (HIV)-associated dementia (HAD) is a frequent complication in HIV+ subjects. Several electrophysiological markers and motor control are altered in HIV+ subjects, including event-related potentials (N2-P3 changes). These are electrophysiological indicators of cognitive processing. The mechanisms by which HIV induces neurophysiological abnormality is still under research. However, several neurotransmitters have been implicated. For example, glutamate and the vasoactive intestinal neuropeptide (VIP). In this study, we support further this notion indicating that HIVgp120, a glycoprotein derived from HIV, is involved in the pathogenesis of neuropsychiatric abnormalities. We also have observations suggesting that one HIVgp120 mechanism of action is to interfere with cholinergic neurotransmission. Our results indicate that event related potentials (ERP) were affected by HIVgp120, in particular N2 and P3. In addition, motor coordination was severely affected. Both parameters were maintained near normality when rats were simultaneously treated with nicotine. These results support further an HIVgp120-caused alteration of cholinergic neurotransmission that might be part of the etiology of neuropsychiatric disturbances. PMID- 16269214 TI - Structural characteristics of a lipid peroxidation product, trans-2-nonenal, that favour inhibition of membrane-associated phosphotyrosine phosphatase activity. AB - Protein-tyrosine phosphatases (PTPs) are very susceptible to oxidation by reactive oxygen species (ROS), which induce the oxidation of catalytic cysteines, thereby inactivating these PTPs. PTPs are also inactivated by treatment with different aldehydes (such as trans-2-nonenal), produced after tissue damage by ROS. However, the molecular mechanisms behind such aldehyde-due inactivation remain unknown. Using commercially available compounds, we examined the structural characteristics of trans-2-nonenal that allow the inhibition of platelet membrane-associated PTP activity, as well as how these compounds affect the dynamics of SH-, CO- and NH2- protein groups on the membranes. PTP was effectively inhibited by physiological amounts of trans-2-nonenal (1-10 microM). Incubation with trans-2-nonene (10 microM) also decreased PTP activity, although to a lower extent. Treatment with nonyl aldehyde almost eliminated PTP inhibition. Decreases in protein thiols were visible after trans-2-nonenal and trans-2-nonene treatments. Both the latter compounds also increased protein carbonyls (although trans-2-nonenal was more effective) and decreased protein amino groups to an equal extent. Collectively, our data indicate that alpha,beta unsaturation (and not a double bond in another position) is the most important structural determinant for PTP inhibition, the alkenal with 9-carbon atoms being the most effective in eliciting such inhibition. The data allow us to predict the modification of sulfhydryls and/or the formation of addition products with lysyl or histidyl residues, and hence the kind of specific antibodies that it would be necessary to generate in order to test such modifications directly. PMID- 16269215 TI - The KRAS oncogene: past, present, and future. PMID- 16269216 TI - Current practice of thromboprophylaxis in the burn population: a survey study of 84 US burn centers. AB - While there is limited prospective data on the incidence of venous thromboembolism (VTE) in the burn population, there are no prospective studies on the efficacy and safety of VTE prophylaxis in these patients. Despite lack of such data, we hypothesized that most burn centers practice some form of prophylaxis. Eighty-four US burn centers were contacted regarding their modality of VTE prophylaxis, if any. Of the 84 US burn centers, 71 were enrolled in this survey. 76.1% centers reported routine VTE prophylaxis. Modalities included sequential compression device (SCD) (33), subcutaneous heparin (31), enoxaparin (13), dalteparin (3), and intravenous heparin infusion (1). Twenty-one reported combined modalities of SCD and subcutaneous heparin (19), SCD and enoxaparin (1), or SCD and dalteparin (1). Survey results underscore the need to definitively establish risk factors for VTE in the burn population and to prospectively define an evidence-based standard of care in prophylaxis for those patients. PMID- 16269217 TI - A three decade analysis of factors affecting burn mortality in the elderly. AB - This study's objective was to identify variables that affect the mortality of elderly burn patients and to assess their changes over time. A retrospective review was conducted on all patients 75 or older (n=201) admitted to a university based burn center between 1972 and 2000. Variables examined were age, sex, TBSA, ABSI, inhalation injury, timing from burn to operative intervention, the number of surgical procedures, the number of pre-morbid conditions, and mortality. There were 95 fatalities. TBSA strongly correlated with mortality (p<0.0001). Adjusting for TBSA and inhalation injury, mortality significantly decreased (p=0.04, odds ratio=0.58). Mortality significantly increased with inhalation injury (p<0.01). Fatality risk increased by 400% with inhalation injury. Absence of inhalation injury was not significant with respect to mortality in the 1970s, however there was a significant decrease (p=0.02) in mortality without an inhalation injury in the 1980s and 1990s. ABSI was strongly predictive of mortality (p<0.0001). On average there was a 200% increase in mortality per unit increase of ABSI. The elderly are 58% less likely to die from burns now as compared to the 1970s. Although mortality rose with increasing TBSA equally in each decade, the absolute risk of mortality decreased over time. This data suggests major strides have been made in burn care, however similar success has not been achieved with inhalation injuries. PMID- 16269219 TI - The relevance of the detection of troponins to the forensic diagnosis of cardiac contusion. AB - The forensic diagnosis of cardiac contusion has hitherto been based mainly on anamnesis, concomitant thoracic injuries and the detection of macroscopic changes to the heart. Parallel histological and serological investigations of the heart specific troponins have been conducted with varying results. This paper aims to show whether heart-specific troponins are suitable as a means of securing the diagnosis in proven cases of cardiac contusion and of determining which of the three heart-specific troponins cTnT, cTnI and cTnC are most significant in serology and histology for postmortem diagnosis. In the study, 25 cases of known cardiac contusion and 11 controls without vital myocardial trauma taken from autopsy material were prospectively investigated. Investigation of the venous serum revealed significant differences in the concentrations of the case and control groups for troponin T (mean value 5.5056 versus 0.4982; p=0.014), for troponin C (mean value 263.9280 versus 68.5640; p=0.001) and for troponin I (mean value 1404.0560 versus 36.1650; p=0.003). In histology there are also significantly different depletions between the groups investigated (cTnT: p=0.002; cTnC: p=0.003; cTnI: p<0.001) taking into account the autolysis time. PMID- 16269218 TI - Biological monitoring of experimental human exposure to hydrocarbon solvent mixtures. AB - The aim of the study was to develop and to validate a suitable analytical method in order to assess the internal exposure of persons to commercial products of hydrocarbon solvent mixtures (HSM). Twenty healthy volunteers were exposed to vapours of five commercial HSM for 8h at 200-1,000 mg/m(3) air. Aromatic-rich, aromatic-poor and aromatic-free HSM were used, as well as isohexane and technical hexane mixtures. A total of 300 exposures were carried out at rest or with an exercise period of 10 min/h at 50 and 75 W. Blood samples for the determination of the HSM were collected before and immediately after exposure. They were analyzed with a headspace analyzer by gas chromatography and mass spectrometry. The analytical method has detection limits of 2-50 microg HSM/L blood. With this method we obtained intra- and interassay variation coefficients of 3.7-15.1%, at concentrations of 53-1,500 microg HSM/L blood. The mean values of the HSM of the 20 volunteers after 8h range between 89 mug/L (technical hexane-mixture) and 1,369 microg/L blood (aromatic-free HSM) at rest. Physical exercises of 50 and 75 W, respectively, lead to a significant increase of the blood-concentrations by mean factors between 1.2 and 1.9 for the five HSM. In conclusion, our results demonstrate that physical activity should be considered in the setting of occupational exposure limits. PMID- 16269220 TI - Separation and determination of the effective components in the alabastrum of Edgeworthia chrysantha Lindl. by micellar electrokinetic capillary chromatography. AB - A rapid and efficient micellar electrokinetic capillary chromatography (MEKC) method was developed to analyze edgeworoside C (1), kaempferol-3-O-beta-D glucoside (2) and rutin (3) in the alabastrum of Edgeworthia chrysantha Lindl. for the first time. The factors that affect the separation were studied, such as the concentrations of the buffer, SDS, and organic modifier, the apparent pH, the applied voltage and temperature. The analytes were well separated within 15 min with an electrolyte containing 25 mM Na(2)B(4)O(7), 30 mM NaH(2)PO(4), 60mM SDS and 15% acetonitrile (pH(*) 9.1) at 25 kV and 15 degrees C. The correlation coefficients between the peak areas of analytes and the corresponding concentrations were 0.9976-0.9981 under the optimum conditions. The relative standard deviations (R.S.D.) of the migration time and peak area were in the range 0.6-1.7 and 1.9-5.3%, respectively. The contents of analytes in E. chrysantha Lindl. collected from the different places were successfully determined with the recoveries ranging from 95.9 to 104.3%. And, the results demonstrated that there was significant difference between the two real samples. PMID- 16269221 TI - Activity of faropenem tested against Neisseria gonorrhoeae isolates including fluoroquinolone-resistant strains. AB - We evaluated the anti-gonococcal potency of faropenem along with 7 comparator reference antimicrobials against a preselected collection of clinical isolates. The 265 isolates were inclusive of 2 subsets: 1) 76 well-characterized resistant phenotypes of gonococcal strains (53 quinolone-resistant strains--31 with documented quinolone resistance-determining region changes from Japan, 15 strains resistant to penicillin and tetracycline, and 8 strains with intermediate susceptibility to penicillin) and 2) 189 recent isolates from clinical specimens in 2004 from 6 states across the United States where quinolone resistance is prevalent. Activity of faropenem was adversely affected by l-cysteine hydrochloride in IsoVitaleX (4-fold increase in [minimal inhibitory concentration] MIC50; 0.06 versus 0.25 microg/mL). The rank order of potency of the antimicrobials for the entire collection was ceftriaxone (MIC90, 0.06 microg/mL) > faropenem (0.25 microg/mL) > azithromycin (0.5 microg/mL) > cefuroxime (1 microg/mL) > tetracycline (2 microg/mL) > penicillin = ciprofloxacin = levofloxacin (4 microg/mL). Using MIC90 for comparison, faropenem was 4-fold more potent than cefuroxime (0.25 versus 1 microg/mL), but was 4-fold less active than ceftriaxone (0.25 versus 0.06 microg/mL). Although the activity of faropenem was not affected by either penicillinase production (MIC90, 0.12 microg/mL, penicillinase-positive) or increasing ciprofloxacin MIC (0.25 microg/mL, ciprofloxacin-resistant), increasing penicillin MIC was associated with an increase in MIC90 values (0.016 microg/mL for penicillin-susceptible to 0.25 microg/mL for penicillin-resistant strains). Among the recent (2004) clinical gonococcal isolates tested, reduced susceptibility to penicillins, tetracycline, and fluoroquinolones was high (28.0-94.2%). Geographic distribution of the endemic resistance rates of gonococci varied considerably, with 16.7-66.7% of the gonococcal isolates being ciprofloxacin-resistant in Oregon, California, Washington, and Hawaii. Faropenem retained its potency against these recent clinical strains and also quinolone-resistant strains from Japan (MIC90, < or =0.25 microg/mL). In summary, the excellent activity of faropenem against the gonococcal strains analyzed irrespective of the resistance phenotype, along with its beta-lactamase stability, makes it an ideal contender for further development as an oral beta-lactam agent to treat uncomplicated gonococcal infections due to strains emerging with resistant to penicillins, tetracyclines, and fluoroquinolones. PMID- 16269222 TI - Evaluation of phenotypic screening methods for detecting plasmid-mediated AmpC beta-lactamases-producing isolates of Escherichia coli and Klebsiella pneumoniae. AB - Detection of plasmid-mediated (P-M) AmpC beta-lactamase-producing isolates is considered critical for epidemiologic studies and hospital infection control, but the documents of the Clinical and Laboratory Standards Institute do not contain any recommendation for the phenotypic detection. In this study, phenotypic detection methods, cefoxitin-Hodge test and induction test, were evaluated using cefoxitin-resistant Escherichia coli and Klebsiella pneumoniae isolates. The cefoxitin-Hodge test detected all bla(CMY-10), and 97.4% of bla(CMY-2) allele positive isolates, but only 57.3% of bla(DHA-1) allele-positive isolates. Induction test with an aztreonam and an amoxicillin-clavulanic acid disk was more sensitive than with cefoxitin disk, which detected 86.6% of bla(DHA-1) allele positive isolates. These phenotypic tests should be useful to screen P-M AmpC beta-lactamase-producing E. coli and K. pneumoniae isolates. PMID- 16269223 TI - Clinical significance of Aerococcus urinae: a retrospective review. AB - A retrospective chart review was performed on 54 patients with positive urine cultures during a 1-year period to assess the clinical significance of Aerococcus urinae. Based on predetermined criteria, patients were classified into 2 groups: those with urinary tract infections (UTIs) and those who were considered colonized. The majority of the patients were > or =65 years old and were female. Only 31% of patients with UTI and 45% of colonized patients had A. urinae isolated in pure cultures. Both groups had significant but similar underlying medical conditions, with urologic conditions being predominant. Significantly more patients in the UTI group had urinary catheters (P < .01). No direct complications or invasive disease was recognized in either group regardless of whether patients were treated with antibiotics. Apparently, A. urinae is a relatively avirulent organism when cultured from urine. PMID- 16269224 TI - A 5'-nuclease real-time reverse transcriptase-polymerase chain reaction assay for the detection of a broad range of influenza A subtypes, including H5N1. AB - A 5'-nuclease real-time reverse transcriptase-polymerase chain reaction assay was developed for the detection of influenza type A and was validated using a range of influenza A subtypes, including avian strains, and 126 nasopharyngeal aspirate samples. The results show the assay is suitable for screening for influenza A infections, particularly in regions where avian strains may be circulating. PMID- 16269225 TI - Sentinel node biopsy for cT1 and cT2a gastric cancer. AB - AIMS: To evaluate the feasibility and accuracy of sentinel node (SN) biopsy for gastric cancer. PATIENTS AND METHODS: One hundred patients with gastric cancer diagnosed as cT1 (n=80) or cT2a (n=20) were enrolled. Indocyanine green-stained SNs were analysed by hematoxylin and eosin staining (n=100) and by cytokeratin immunohistochemistry (n=50). RESULTS: SNs were identified in 94 of the 100 patients and the mean number of SNs was 4.4 (range, 1-12). Of these 94 patients, 14 patients had lymph node metastases. Two patients with T1 and one patient with T2 had metastases in non-SNs alone by hematoxylin and eosin staining (diagnostic accuracy =97.3% in T1 and 95.0% in T2). All three patients with a false negative result had a tumour, which was more than 4 cm in size and signet ring cell histology. In two of them, the tumour was located at lesser curvature. By immunohistochemical staining, three patients with T1 and one patient with T2 were found to have lymph node micrometastases in non-SNs alone among 45 patients (diagnostic accuracy =92.1% in T1, 85.7% in T2). CONCLUSION: SN biopsy using indocyanine green can be performed rapidly and easily with a high detection rate and accuracy in patients with T1 gastric cancer. However, it should be performed with caution for large tumours with a signet ring cell histology located at lesser curvature due to the possibility of a false negative result. PMID- 16269226 TI - Positron emission tomography in patients with breast cancer using (18)F-3'-deoxy 3'-fluoro-l-thymidine ((18)F-FLT)-a pilot study. AB - BACKGROUND: This pilot study investigated the feasibility of (18)F-3'-deoxy-3' fluoro-l-thymidine ((18)F-FLT) as a positron emission tomography (PET) tracer for the visualisation of breast cancer. METHODS: Patients with breast cancer underwent (18)F-FLT-PET prior to surgery. The uptake of (18)F-FLT was determined in the primary tumour and in the axilla. RESULTS: Eight tumours were visualized by (18)F-FLT-PET with a mean uptake value (SUV(mean)) of 1.7 and mean tumour-non tumour ratio (TNT) of 5.0. In seven patients, axillary lymph-node metastases were found at pathological examinations, however, (18)F-FLT-PET showed uptake in only two large (and clinically evident) lymph-node metastases. CONCLUSIONS: (18)F-FLT shows uptake in most primary breast tumours and in large axillary lymph-node metastases. PMID- 16269227 TI - Predictors of tumour involvement in remaining axillary lymph nodes of breast cancer patients with positive sentinel lymph node. AB - AIMS: To characterize the various clinicopathologic features in cases of breast cancer with positive sentinel lymph nodes (SLNs), in order to determine factors that might help in predicting the involvement of the non-SLNs. METHODS: A retrospective database review was performed of 726 breast cancer patients with stage 0-II, in whom SLNs were successfully identified. One hundred eighty-five of these patients showed positive SLNs, and subsequently underwent axillary lymph node dissection (ALND). These cases were divided into two groups based on the presence or absence of metastases in the non-SLNs, i.e. positive non-SLNs (NSLN+; 81 cases) and negative non-SLNs (NSLN-; 104 cases). RESULTS: Multivariate analysis revealed that a larger size of the primary tumour (>2.0cm), presence of lymphatic invasion, larger size of the largest SLN metastasis (>2mm), and a 100% metastatic rate in the SLNs (number of positive SLNs/number of harvested SLNs) were significantly associated with NSLN+. Among the cases in which all the four factors were present, 73% (30/41) were found to have NSLN+. CONCLUSION: We found four independent predictors in relation to non-SLN metastasis. Although these factors might be useful for determining the need of additional ALND, it would seem that even the presence of all of these four factors in combination may be insufficient to safely omit ALND. Thus, until further evidence is accumulated from the results of large clinical trials, ALND would still be recommended for patients with SLN metastasis. PMID- 16269229 TI - [Sciatic nerve block]. PMID- 16269228 TI - [National survey in France about continuing medical education among anaesthesiologists]. AB - STUDY DESIGN: The aim of this study was to assess how French anaesthesiologists perform continuing medical education (CME). METHODS: A 73 items survey was mailed to 1,000 anaesthesiologists (11% of anaesthesiologists population) of geographic, gender and institution representative of national anaesthesiologist demography. A second mail was sent to non-responders, 6 weeks later. RESULTS: The answer rate was 40.8% and the sample of responders appeared to match the French anaesthesiologist population. Seventy-two percent of surveyed anaesthesiologists were affiliated to the French society of anaesthesia-intensive care and 24% to other medical societies. Attendance to French congresses was 81% and miscellaneous congresses were equally appreciated in terms of quality. Only 17% of surveyed anaesthesiologists attended international congresses. Multi-thematic congresses were preferred by 67%. The annual time devoted to congresses was 6 days (median) with additional 4 days (median) reserved for practical courses. French medical journals and international journals had a reading rate of 89 and 37%, respectively. For 61% of responders CME was funded by institutional grants. Internet CME use was found in 73% of anaesthesiologists. Time and money were the two most frequent reasons invoked for CME restriction. CONCLUSION: CME is a broadly shared activity, which still remains focussed on national resources. PMID- 16269230 TI - [Fat pulmonary embolism after liposuction]. AB - A 24-year-old woman undergoes buttock's liposuction as an outpatient procedure. As she went back home, progressive dyspnea, respiratory distress and collapse developed. At hospital admission, she was dyspneic with thoracic oppression, tachycardia and anguish. Chest X-ray and thoracic CT scan suggested a pulmonary localisation of fat emboli. Symptomatic treatment allowed complete recovery. This report discusses diagnosis of fat emboli after liposuction as well as epidemiology and physiopathology. PMID- 16269231 TI - [Anaesthesia and critical care for scheduled infrarenal abdominal aortic aneurysm surgery]. AB - OBJECTIVES: Patient scheduled for infrarenal abdominal aortic aneurysm surgery carries a high risk of cardiac or respiratory comorbidity. To outline the perioperative management for these patients. METHODS: Review of the literature using MesH Terms "abdominal aortic aneurysm", "anesthesia", "analgesia" "critical care" and/or "surgery" in Medline database. RESULTS: Cardiac preoperative evaluation and management have recently been reviewed. Intermediate and high-risk patients should undergo non-invasive cardiac testing to decide between a preoperative medical strategy (using betablocker+/-statin and aspirin) and an interventional strategy (coronary angioplasty or cardiac surgery). Perioperative myocardial ischaemia should also be investigated by clinical, electrocardiographic and biologic monitoring such as plasmatic troponin Ic dosage. Specific score could also assess the respiratory failure risk preoperatively. Epidural analgesia decreases this risk. There is no evidence that a pharmacological treatment decreases the incidence of acute renal failure after aortic surgery. Endovascular repair is actually recommended for older, higher risk patients or patients with a hostile abdomen or other technical factors that may complicate standard open repair. PMID- 16269232 TI - [Evaluation of a continuous training program at Bichat hospital for in-hospital cardiac arrest resuscitation]. AB - Management of in-hospital cardiac arrest is now considered as a hospital quality indicator. Such management actually requires training health care workers (HCWs) for basic life support (BLS). OBJECTIVE: To assess the usefulness and efficacy of a short mandatory BLS training course amongst general ward HCWs in a 1,200 bed teaching hospital. STUDY DESIGN: The in-hospital medical emergency team (MET) established a 45-min BLS training course comprising 10 goals for basic CPR and preparing for the arrival of the MET. Assessment was based on satisfaction questionnaires, cross-sectional evaluation of knowledge and skills of HCWs before and 1 year after the start of the training course. Efficacy of BLS performed on ward was assessed by the MET on scene. RESULTS: One year after, 68 training sessions had been fulfilled and 522 HCWs had been trained (46.27% of total HCWs). HCWs were satisfied with the teaching course. Instant retention of objectives was over 90%. Cross-sectional surveys showed an improvement of BLS knowledge and skills. The knowledge of initial clinical assessment remained low. Knowledge and skills were significantly higher amongst HCWs who had been trained than amongst those who had not. Unfortunately, general ward BLS performance showed no improvement. CONCLUSION: Short mandatory training courses are stimulating and well appreciated amongst HCWs. Although basic knowledge and skills improve dramatically, no improvement of on-scene BLS performance occurs. PMID- 16269233 TI - [Anaesthesia for a non cardiac surgery in a patient with an Eisenmenger syndrome. Interest of a non invasive haemodynamic monitoring]. AB - Patients with an Eisenmenger syndrome have an instuble hemodynamic status. During a general anaesthesia, the intracardiac shunt has to maintain the correct orientation and volume, adapted to each patient, in such a condition, to avoid the risk of hypoxemia and cardiac failure. The haemodynamic monitoring with a Swan Ganz catheter could be useful. But it is necessary to evaluate the advantage and the risks when the technique is used in these pathological circumstances. Moreover, when the cardiac output is measured with the thermodilution technique, the right-left intra cardiac shunt volume, is not taking into account. The continuous haemodynamic monitoring, with a simplified transoesophageal echo Doppler system, as it was done in this case, allows appreciate the real quantitative variations of the shunt. In this way the more adequate calculation of some others haemodynamic parameters, over all the total systemic vascular resistances, allows a more precise therapeutic approach. PMID- 16269234 TI - The HSP90 family of genes in the human genome: insights into their divergence and evolution. AB - HSP90 proteins are important molecular chaperones. Transcriptome and genome analyses revealed that the human HSP90 family includes 17 genes that fall into four classes. A standardized nomenclature for each of these genes is presented here. Classes HSP90AA, HSP90AB, HSP90B, and TRAP contain 7, 6, 3, and 1 genes, respectively. HSP90AA genes mapped onto chromosomes 1, 3, 4, and 11; HSP90AB genes mapped onto 3, 4, 6, 13 and 15; HSP90B genes mapped onto 1, 12, and 15; and the TRAP1 gene mapped onto 16. Six genes, HSP90AA1, HSP90AA2, HSP90N, HSP90AB1, HSP90B1 and TRAP1, were recognized as functional, and the remaining 11 genes were considered putative pseudogenes. Amino acid polymorphic variants were detected for genes HSP90AA1, HSP90AA2, HSP90AB1, HSP90B1, and TRAP1. The structures of these genes and the functional motifs and polymorphic variants of their proteins were documented and the features and functions of their proteins were discussed. Phylogenetic analyses based on both nucleotide and protein data demonstrated that HSP90(AA+AB+B) formed a monophyletic clade, whereas TRAP is a relatively distant paralogue of this clade. PMID- 16269235 TI - DNA microarray-based detection of nosocomial pathogenic Pseudomonas aeruginosa and Acinetobacter baumannii. AB - Infection by nosocomial pathogenic bacteria is increasingly becoming a major threat to the patients in the hospital. We have developed a diagnostic DNA microarray for the detection of two important nosocomial pathogens, Pseudomonas aeruginosa and Acinetobacter baumannii. The diagnostic DNA microarray contains the species-specific probes of 15mer oligonucleotides designed based on the sequences of 23S ribosomal DNA. The performance of DNA microarray in diagnosing P. aeruginosa and A. baumannii was evaluated using reference bacteria as well as clinical specimens such as blood, stool, pus, sputum, urine and cerebrospinal fluid. Using this DNA microarray, A. baumannii could be successfully detected in 11 out of 13 clinical specimens, thus giving the sensitivity of 84.6% with the specificity of 100% and the positive predictive value of 100%. P. aeruginosa could also be detected in 25 out of 26 clinical specimens, showing the sensitivity of 96.2%, the specificity of 100%, and the positive predictive value of 100%. These results suggest that two nosocomial pathogens, P. aeruginosa and A. baumannii, can be efficiently diagnosed by using the DNA microarray developed in this study. PMID- 16269236 TI - Fears, hyperacusis and musicality in Williams syndrome. AB - The study investigated the prevalence of fear and hyperacusis and the possible connections between fear, hyperacusis and musicality in a Swedish sample of individuals with Williams syndrome (WS). The study included 38 individuals and a cross-sectional design, with no matched control group. Two persons, who knew the participant well, completed a questionnaire. On reported fears, 58% of the participants scored higher than +2S.D., compared to a psychometric study. Thirteen percent scored above the suggested cut-off for hyperacusis, compared to 2.5% in a psychometric study. Female participants generally had higher reported fears and hyperacusis compared to male participants. There were also startling findings of correlations between reported fears and hyperacusis. This preliminary report supports a hypothesis that fears and anxiety could be associated with hyperacusis in the WS population. A hypothesis that musicality could serve as a protective factor and prevent anxiety, received no or very limited support. A hypothesis that hyperacusis could be connected to a general, readily arousal, tendency in the sympathetic nervous system and could be seen as vulnerability for psychopathology is discussed. PMID- 16269237 TI - Interpreting hippocampal function as recoding and forecasting. AB - A model of hippocampal function, centered on region CA3, reproduces many of the cognitive and behavioral functions ascribed to the hippocampus. Where there is precise stimulus control and detailed quantitative data, this model reproduces the quantitative behavioral results. Underlying the model is a recoding conjecture of hippocampal computational function. The expanded conjecture includes a special role for randomization and, as recoding progresses with experience, the occurrence of sequence learning and sequence compression. These functions support the putative higher-order hippocampal function, i.e. production of representations readable by a linear decoder and suitable for both neocortical storage and forecasting. Simulations confirm the critical importance of randomly driven recoding and the neurocognitive relevance of sequence learning and compression. Two forms of sequence compression exist, on-line and off-line compression: both are conjectured to support neocortical encoding of context and declarative memory as described by . PMID- 16269238 TI - Digital staining for multispectral images of pathological tissue specimens based on combined classification of spectral transmittance. AB - In this study, the digital transformation (digital staining) of the 16-band multispectral image of a hematoxylin and eosin (HE) stained pathological specimen to its Masson's trichrome (MT) stained counterpart is addressed. The digital staining procedure involves the classification of the various H&E-stained tissue components and then the transformation of their transmittance spectra to their equivalent MT-stained transmittance configurations. Combination of transmittance classifiers were designed to classify the various tissue components found in the multispectral images of an HE-stained specimen, e.g. nucleus, cytoplasm, red blood cell (RBC), fibrosis, etc.; while pseudo-inverse method was used to obtain the transformation matrices that would translate the transmittance spectra of the classified HE-stained multispectral pixels to their MT-stained configurations. To generate the digitally stained image, weighting factors, which were based on the classifiers beliefs, were introduced to the generated transformation matrices. Initial results of our experiments on liver specimens show the viability of multispectral imaging (MSI) to implement a digital staining framework in the pathological context. PMID- 16269239 TI - alpha-[11C]Methyl-L-tryptophan trapping in the orbital and ventral medial prefrontal cortex of suicide attempters. AB - Low serotonin neurotransmission is thought to increase vulnerability to suicidal behavior. To test this hypothesis, we measured brain regional serotonin synthesis, as indexed by PET and alpha-[(11)C]methyl-L-tryptophan trapping, in 10 patients who had made a high-lethality suicide attempt and 16 healthy controls. Compared to healthy controls, suicide attempters had reduced normalized alpha [(11)C]methyl-L-tryptophan trapping in orbital and ventromedial prefrontal cortex. alpha-[(11)C]Methyl-L-tryptophan trapping in these regions correlated negatively with suicide intent. Low serotonin synthesis in the prefrontal cortex might lower the threshold for suicidal behavior. PMID- 16269240 TI - Antiplasmodial activity of ferrocenyl chalcones: investigations into the role of ferrocene. AB - A series of ferrocenyl chalcones were synthesized and evaluated in vitro against Plasmodium falciparum (K1) in a [3H] hypoxanthine uptake assay. Appropriate size, electronic, lipophilic and electrochemical parameters were determined for QSAR analysis. The results showed that the location of ferrocene influenced the ease of oxidation of Fe2+ in ferrocene and the polarity of the carbonyl linkage. These parameters were found to influence antiplasmodial activity. A general trend was noted in which compounds with ferrocene adjacent to the carbonyl linkage (series A) were associated with more selective and potent antiplasmodial activities. These compounds had polarized carbonyl linkages, lower lipophilicities and ferrocene rings that were less readily oxidized. The most active analogue was 1 ferrocenyl-3-(4-nitrophenyl)prop-2-en-1-one (28) (IC50 4.6 microM, selectivity index 37 against KB3-1 cells). To understand how the redox properties of ferrocene might influence antiplasmodial activity, the oxidant properties of selected compounds were investigated in antioxidant (ABTS+) and EPR experiments. The incorporation of ferrocene in the chalcone template was found to enhance its role in processes that involved the quenching and generation of free radicals. Thus, ferrocene may participate in redox cycling and this process may contribute to the antiplasmodial activity of ferrocenyl chalcones. However, the extent to which this property is manifested is also influenced by other physicochemical properties (lipophilicity, polarity, and planarity) of the compound. PMID- 16269241 TI - Effect of pre-composting on vermicomposting of kitchen waste. AB - The aim of this work was to test combination of the thermocomposting and vermicomposting to improve the treatment efficiency and assess the optimum period required in each method to produce good quality compost. The results showed that pre-thermocomposting improved vermicomposting of kitchen waste. A 9-day thermocomposting prior to vermicomposting helped in mass reduction, moisture management and pathogen reduction. PMID- 16269242 TI - Soda-anthraquinone, kraft and organosolv pulping of holm oak trimmings. AB - The operating conditions for an organosolv (ethyleneglycol) and two alkaline (soda-anthraquinone and kraft) processes for obtaining cellulose pulp and paper from holm oak (Quercus ilex) wood trimmings were optimized. A range of variation for each process variable (viz. temperature, cooking time and soda or ethyleneglycol concentration) was established and a central composite experimental design involving three independent variables at three different variation levels was applied. The results obtained with the three cooking processes used were compared and those provided by the kraft process were found to be the best. Thus, the tensile index values it provided (5.9-16.3 N m/g) were 23.7% and 41.5% better than those obtained with the soda-AQ and ethyleneglycol processes, respectively. Also, the kraft process provided the best burst index, brightness and kappa number values. Based on the optimum working ranges, the temperature and cooking time were the variables resulting in the most and least marked changes, respectively, in pulp properties. PMID- 16269244 TI - Postural visual dependence after recent stroke: assessment by optokinetic stimulation. AB - INTRODUCTION: This study was conducted to assess visual dependence of postural control early after stroke. DESIGN: Case control study. SETTING: A Physical Medicine and Rehabilitation Department. METHOD: Twenty-five patients impaired by severe hemiplegia due to a recent first stroke, were examined. Fourteen had a right and 11 a left hemispheric lesion. There were aged 52+/-13 years, time since stroke was 30+/-12 days. Patients' data were compared to data for 25 healthy subjects. Sitting posture was assessed on a dynamic balance, using two parameters: frontal plane displacement of the centre of pressure under optokinetic stimulation (OKS), and the total length of centre of pressure displacement for the stability reaction. On the basis of 90th percentile control data, patients' behaviour was classified as totally visuo independent (VI), totally visuo dependent (VD) or mixed. RESULTS: Body tilt under OKS was greater in patients than controls. No control subject was totally VD, 19 subjects were totally VI. Four patients were totally VD and only six were VI. The only clinical parameter linked to the effect of OKS was the sensitivity impairment. Overall patients with visuospatial neglect were the most perturbed, but two were totally visuo independent. DISCUSSION AND CONCLUSION: Assessment of postural variations in sitting under OKS is proposed for estimating visual dependence early after stroke. Individual reactions are more important than mean group reactions. Visual dependence is not solely due to neurological impairment, implying that previous physiological behaviour may be involved. Knowledge of these characteristics may affect rehabilitation programmes. PMID- 16269245 TI - Glucose as a regulator of eukaryotic gene transcription. AB - Glucose has essential metabolic roles as both a fuel for energy and a substrate for the biosynthesis of cell components. Because of its central importance, many cells have evolved mechanisms to sense glucose levels in their environment and to adapt the expression of their genetic information to glucose availability. This glucose signaling is vital in mammalian cells where derangements in glucose utilization might contribute to conditions such as obesity and type 2 diabetes. Two crucial issues stand out in understanding pathways of glucose-regulated gene transcription. First, how do cells sense changing glucose levels? Second, how is this signal transduced to the transcriptional apparatus of the cell? In mammalian cells, glucose sensing involves the detection of changes in glucose metabolism rather than glucose itself. A transcription factor that is involved in mediating responses to glucose, ChREBP, has been identified recently and studies have begun to elucidate the molecular basis of coupling between glucose metabolism and transcription factor activity. PMID- 16269246 TI - The endocrine function of the heart. AB - Atrial cardiocytes in the heart of mammals produce in a regulated manner the polypeptide hormones atrial natriuretic factor (ANF, ANP) and brain natriuretic peptide (BNP). The biological actions of ANF and BNP are similar; they include the modulation of systems that tend to increase extracellular fluid volume and blood pressure, such as the renin-angiotensin system and the sympathetic nervous system. Additionally, both hormones have potent growth-regulating properties. ANF and BNP signal by activating membrane-bound guanylyl cyclase receptors, leading to an increase in intracellular cGMP and thus affecting the activity of cGMP regulated enzymes and ion channels. Under chronic hemodynamic overload, cardiac ANF and BNP synthesis and secretion are increased. This increase is viewed as a cardioprotective mechanism, given the beneficial effects of ANF and BNP on cardiac preload, afterload and cardiovascular growth. As discussed in this review, some basic facts regarding the synthesis and secretion of ANF and BNP and their peripheral effects remain to be clarified. Nevertheless, at the clinical level, the elevation of circulating ANF and BNP in heart failure or following acute coronary syndromes has been shown to have diagnostic and prognostic implications. Moreover, these peptides themselves hold promise as therapeutic agents in the treatment of heart failure. Additional pharmaceutical applications might be gleaned from current preclinical and clinical studies showing beneficial effects of ANF or BNP in the treatment of hypertension, bronchospasm and in tissue remodeling following acute myocardial infarction. PMID- 16269247 TI - A novel esterase from Bacillus subtilis (RRL 1789): purification and characterization of the enzyme. AB - An esterase (EC 3.1.1.1) produced by an isolated strain of Bacillus subtilis RRL 1789 exhibited moderate to high enantioselectivity in the kinetic resolution of several substrates like aryl carbinols, hydroxy esters, and halo esters. The enzyme named as B. subtilis esterase (BSE), was purified to >95% purity with a specific activity of 944 U/mg protein and 12% overall yield. The purified enzyme is approximately 52 kDa monomer, maximally activity at 37 degrees C and pH 8.0 and fairly stable up to 55 degrees C. The enzyme does not exhibit the phenomenon of interfacial activation with tributyrin and p-nitrophenyl butyrate beyond the saturation concentration. The enzyme showed preference for triacyglycerols and esters of p-nitrophenol with short chain fatty acid. Presence of Ca2+ ions increases the activity of enzyme by approximately 20% but its presence does not have any influence on the thermostability of the enzyme. The enzyme is not a metalloprotein and belongs to the family of serine proteases. The N-terminal amino acid sequence of BSE determined, as Met-Thr-Pro-Glu-Iso-Val-Thr-Thr-Glu-Tyr Gly- revealed similarity with the N-terminal amino acid sequence of p nitrobenzylesterase of B. subtilis. PMID- 16269248 TI - Correspondence between EEG-fMRI and EEG dipole localisation of interictal discharges in focal epilepsy. AB - EEG-fMRI and EEG dipole source localisation are two non-invasive imaging methods that can be applied to the study of the haemodynamic and electrical consequences of epileptic discharges. Using them in combination has the potential to allow imaging with the spatial resolution of fMRI and the temporal resolution of EEG. However, although considerable data are available concerning their concordance in studies involving event-related potentials (ERPs), less is known about how well they agree in epilepsy. To this end, 17 patients were selected from a database of 57 who had undergone an EEG-fMRI scanning session followed by a separate EEG session outside of the scanner. Spatiotemporal dipole modelling was compared with the peak and closest EEG-fMRI activations and deactivations. On average, the dipoles were 58.5 mm from the voxel with the highest positive t value and 32.5 mm from the nearest activated voxel. For deactivations, the corresponding values were 60.8 and 34.0 mm. These values are considerably higher than is generally observed with ERPs, probably as a result of the relatively widespread field, which can lead to artificially deep dipoles, and the occurrence of EEG-fMRI responses remote from the presumed focus of the epileptic activity. The results suggest that EEG and MEG inverse solutions for equivalent current dipole approaches should not be strongly constrained by EEG-fMRI results in epilepsy, and that the use of distributed source modelling will be a more appropriate way of combining EEG-fMRI results with source localisation techniques. PMID- 16269249 TI - Interrupting the "stream of consciousness": an fMRI investigation. AB - In functional neuroimaging, a local decrease in blood flow during an active task, relative to a "resting" baseline, is referred to as task-induced deactivation (TID). TID may occur when resources shift from ongoing, internally generated processing typical of "resting" states to processing required by an exogenous task. We previously found specific brain regions in which TID increased as task processing demands increased. When engaged in an exogenous cognitive task, reallocation of resources from areas involved in internal processing should result in suspension of that processing. Self-reported thought content has been used as an indicator of the extent of internal processing activity. We investigated the relationship between TID and task-unrelated thought (TUT) frequency using an auditory target detection task with seven levels of task difficulty. At varied intervals during task performance, subjects indicated whether they were experiencing a TUT. We expected TUT frequency to decrease as task demands increased and for this pattern to correlate with TID magnitude across conditions. Generally, fewer TUTs were reported during difficult task conditions than during easier conditions. As TID magnitude increased across task conditions, the frequency of TUTs declined (r = 0.90, P = 0.005). Four left hemisphere regions (posterior parieto-occipital cortex, anterior cingulate gyrus, fusiform gyrus, and middle frontal gyrus) showed strong relationships between TUTs and TID (r > 0.79, P < 0.05 corrected). As these regions have been implicated in semantic processing and self-referential thought, the findings support the suspension of internal cognitive processing as one mechanism for TID. PMID- 16269251 TI - An fMRI study of the motor system in patients with neuropsychiatric systemic lupus erythematosus. AB - Functional cortical changes have been demonstrated in patients with several neurological conditions, including stroke, tumors and MS. The correlation found between the extent of fMRI activations and the extent and severity of brain structural damage suggests an adaptive role of these functional changes. In this study, we assess, using fMRI, the brain pattern of movement-associated cortical activations in neuropsychiatric systemic lupus erythematosus (NPSLE) patients and investigate whether the extent of cortical reorganization is associated with the extent of brain pathology, measured on dual-echo and diffusion tensor (DT) MR images. From 14 right-handed NPSLE patients and 14 matched controls, we obtained: (a) fMRI during the performance of repetitive flexion-extension of the last four fingers of the right hand; (b) dual-echo and (c) pulsed-gradient spin-echo echo planar sequence to calculate DT MRI maps of the normal-appearing white (NAWM) and gray (NAGM) matter. Brain T2-visible abnormalities were detected in 11 NPSLE patients. Compared with controls, NPSLE patients had significantly higher NAWM fractional anisotropy histogram peak height (P = 0.005), and more significant activations of the contralateral primary sensorimotor cortex, putamen and dentate nucleus. They also had more significant activations of several regions located in the frontal and parietal lobes as well as of MT/V5 and the middle occipital gyrus, bilaterally. Strong correlations (r values ranging from 0.79 to 0.87) were found between relative activations of sensorimotor areas and the extent and severity of brain damage. Movement-associated functional cortical changes do occur in patients with NPSLE and might contribute to the maintenance of their normal functional capacities. PMID- 16269250 TI - Cortical reconstruction using implicit surface evolution: accuracy and precision analysis. AB - Two different studies were conducted to assess the accuracy and precision of an algorithm developed for automatic reconstruction of the cerebral cortex from T1 weighted magnetic resonance (MR) brain images. Repeated scans of three different brains were used to quantify the precision of the algorithm, and manually selected landmarks on different sulcal regions throughout the cortex were used to analyze the accuracy of the three reconstructed surfaces: inner, central, and pial. We conclude that the algorithm can find these surfaces in a robust fashion and with subvoxel accuracy, typically with an accuracy of one third of a voxel, although this varies with brain region and cortical geometry. Parameters were adjusted on the basis of this analysis in order to improve the algorithm's overall performance. PMID- 16269252 TI - Phylogenetic relationships of South American lizards of the genus Stenocercus (Squamata: Iguania): A new approach using a general mixture model for gene sequence data. AB - The South American iguanian lizard genus Stenocercus includes 54 species occurring mostly in the Andes and adjacent lowland areas from northern Venezuela and Colombia to central Argentina at elevations of 0-4000m. Small taxon or character sampling has characterized all phylogenetic analyses of Stenocercus, which has long been recognized as sister taxon to the Tropidurus Group. In this study, we use mtDNA sequence data to perform phylogenetic analyses that include 32 species of Stenocercus and 12 outgroup taxa. Monophyly of this genus is strongly supported by maximum parsimony and Bayesian analyses. Evolutionary relationships within Stenocercus are further analyzed with a Bayesian implementation of a general mixture model, which accommodates variability in the pattern of evolution across sites. These analyses indicate a basal split of Stenocercus into two clades, one of which receives very strong statistical support. In addition, we test previous hypotheses using non-parametric and parametric statistical methods, and provide a phylogenetic classification for Stenocercus. PMID- 16269253 TI - Neuropsychological tests with lateralizing value in patients with temporal lobe epilepsy: reconsidering material-specific theory. AB - PURPOSE: To assess the ability of neuropsychological tests to determine the side of seizure onset for preoperative assessment in patients with drug-resistant temporal lobe epilepsy. METHODS: Twenty-nine consecutive patients diagnosed with temporal lobe epilepsy (TLE), in whom the epileptogenic focus was clearly identified and localized to either the right or left hemisphere. Patients underwent a full neuropsychological assessment as part of their pre-surgical investigation, including the Boston Naming Test (BNT) and a variety of Wechsler Adult Intelligence Scale-Third Edition (WAIS-III) and Wechsler Memory Scale-Third Edition (WMS-III) subtests. Two multivariate analyses of variance were carried out to assess differences on memory and language measures between groups according to side of epileptogenic focus. Binary logistic regression analysis was performed to find the sets of tests that best predicted the side of seizure onset (determined by EEG and MRI). RESULTS: Memory multivariate analysis of variance failed to show significant differences between the right- and left-sided groups. Among language measures, only the BNT revealed significant differences between the groups. The neuropsychological measures that best predicted the side of seizure onset were the BNT and Visual Reproduction II. CONCLUSIONS: Language measures predict the side of seizure focus better than memory measures. The results of this study in a sample of drug-resistant temporal lobe epilepsy patients challenge the memory material-specific theory for the side of seizure focus. PMID- 16269254 TI - Gadolinium-enhanced magnetic resonance angiography, colour duplex and digital subtraction angiography of the lower limb arteries from the aorta to the tibio peroneal trunk in patients with intermittent claudication. AB - OBJECTIVES: To evaluate the sensitivity, specificity, positive and negative predictive value of contrast-enhanced (gadolinium) magnetic resonance imaging (CE MRA) and colour duplex ultrasound (CDU) of lower limb arteries. DESIGN: Prospective, single centre study. MATERIAL AND METHODS: A consecutive series of 58 patients with intermittent claudication (IC) were examined with CE-MRA and CDU from the infrarenal aorta to the tibio-peroneal trunk with digital subtraction angiography (DSA) as reference. The arterial tree was divided into 15 segments, pooled into three regions; suprainguinal, thigh and knee. Sensitivity, specificity, positive and negative predictive values for significant obstructions were calculated. Cohen Kappa statistics was used to establish agreement between the three methods. RESULTS: The sensitivity (specificity in parentheses) for significant obstructions in the suprainguinal region were 96% (94%) for CE-MRA and 91% (96%) for CDU, in the thigh region 92% (95%) for CE-MRA and 76% (99%) for CDU, and in the knee region 93% (96%) for CE-MRA and 33% (98%) for CDU. CDU failed to visualize 10% of suprainguinal, 2% of thigh and 13% of knee-region arterial segments. CONCLUSIONS: Both CE-MRA and CDU are good alternatives to DSA in the suprainguinal- and thigh-region. In the knee region only CE-MRA can be relied upon as an alternative to DSA. Imaging by CDU is not suited to situations were evaluation of runoff vessels is important. PMID- 16269256 TI - Staged endovascular stent-grafting and surgical treatment of a secondary aortoduodenal fistula. AB - Secondary aortoenteric fistula is a dramatic, rather infrequent late complication occurring mostly after abdominal aortic surgery. Currently, graft excision and in situ bypass is considered the treatment of choice, but it is associated with significant mortality and morbidity. Herein, we describe the case of a secondary aortoduodenal fistula treated by staged endovascular stent-grafting and surgical closure of the fistula. Forty days after stent-grafting, Tc-99m-HMPAO labelled leukocyte scanning failed to identify leukocyte infiltration of the graft and there were no clinical signs of infection. At 8-month follow up, the patient was asymptomatic. PMID- 16269257 TI - Association of cardiovascular risk factors with pattern of lower limb atherosclerosis in 2659 patients undergoing angioplasty. AB - OBJECTIVES: Aim of this study is to correlate distribution pattern of lower limb atherosclerosis with cardiovascular risk factor profile of patients with peripheral arterial occlusive disease (PAD). PATIENTS AND METHODS: Analysis is based on a consecutive series of 2659 patients (1583 men, 1076 women, 70+/-11 years) with chronic PAD of atherosclerotic origin undergoing primary endovascular treatment of lower extremity arteries. Pattern of atherosclerosis was grouped into iliac (n=1166), femoropopliteal (n=2151) and infrageniculate (n=888) disease defined according to target lesions treated. A multivariable multinomial logistic regression analysis was performed to assess relation with age, gender and classical cardiovascular risk factors (diabetes mellitus, arterial hypertension, hypercholesterolemia, cigarette smoking) using femoropopliteal disease as reference. RESULTS: Iliac disease was associated with younger age (RRR 0.95 per year of age, 95%-CI 0.94-0.96, p<0.001), male gender (RRR 1.32, 95%-CI 1.09-1.59, p=0.004) and cigarette smoking (RRR 2.02, 95%-CI 1.68-2.42, p<0.001). Infrageniculate disease was associated with higher age (RRR 1.02, 95%-CI 1.01 1.02, p<0.001), male gender (RRR 1.23, 95%-CI 1.06-1.41, p=0.005) and diabetes mellitus (RRR 1.68, 95%-CI 1.47-1.92, p<0.001). Hypercholesterolemia was less prevalent in patients with lesions below the knee (RRR 0.82, 95%-CI 0.71-0.94, p=0.006), whereas no distinct pattern was apparent related to arterial hypertension. CONCLUSION: Clinical phenotype of peripheral atherosclerosis varies with prevalence of cardiovascular risk factors suggesting differences in mechanisms involved in iliac as compared with infrageniculate lesions. Identification of molecular mechanism might have influence on future therapeutic strategies in PAD patients. PMID- 16269258 TI - Analysis of the UDP-glucuronosyltransferase gene in Portuguese patients with a clinical diagnosis of Gilbert and Crigler-Najjar syndromes. AB - We describe the molecular study in a cohort of 120 Portuguese patients with the clinical diagnosis of Gilbert syndrome and in one with the diagnosis of Crigler Najjar syndrome type II, as well as a prenatal diagnosis of Crigler-Najjar syndrome type I. Among the 120 unrelated patients with Gilbert syndrome, 110 were homozygous for the [TA]7 allele ([TA]7/[TA]7), and one patient was a compound heterozygote for two different insertions ([TA]7/[TA]8). The remaining 9 patients were heterozygous for the TA insertion ([TA]6/[TA]7). Additional studies in these 9 patients revealed heterozygosity for the c.674T>G, c.488_491dupACCT and c.923G>A mutations, in 1, 1 and 4 patients, respectively. The patient with Crigler-Najjar syndrome type II was a compound heterozygote for [TA]7 and the c.923G>A mutation. The undocumented polymorphisms c.-1126C>T and c.997-82T>C were also detected in the course of this study. Prenatal diagnosis in a family with a boy previously diagnosed as Crigler-Najjar syndrome type I and homozygosity for the c.923G>A mutation revealed that the fetus was unaffected. Homozygosity for the [TA] insertion was found to be the most frequent cause of GS in our population. Identification of further mutations in the UGT1A1 gene was also seen to contribute significantly towards diagnosis. PMID- 16269259 TI - Opsoclonus-myoclonus in an HIV-infected child on antiretroviral therapy--possible immune reconstitution inflammatory syndrome. AB - The exact immunopathogenesis and neuroanatomical localization of opsoclonus myoclonus ataxia syndrome remains unclear. We describe a 1 year 9 month old girl who, shortly after commencement of highly active antiretroviral therapy developed opsoclonus-myoclonus syndrome and subsequently died of disseminated cytomegalovirus infection. We postulate on the etiological factors that may have played a role in the disease pathogenesis of the patient's opsoclonus-myoclonus ataxia. Immune reconstitution inflammatory syndrome was considered the most likely because of the initial CD4 depletion and the onset of symptoms shortly after initiation of antiretroviral therapy. Single photon emission computed tomography (SPECT) proved helpful by localizing the area of dysfunction to the cerebellar vermis. PMID- 16269260 TI - Hadamard frequency-encoded SOFAST-HMQC for ultrafast two-dimensional protein NMR. AB - We demonstrate the feasibility of recording (1)H-(15)N correlation spectra of proteins in only one second of acquisition time. The experiment combines recently proposed SOFAST-HMQC with Hadamard-type (15)N frequency encoding. This allows site-resolved real-time NMR studies of kinetic processes in proteins with an increased time resolution. The sensitivity of the experiment is sufficient to be applicable to a wide range of molecular systems available at millimolar concentration on a high magnetic field spectrometer. PMID- 16269261 TI - Application of k- and q-space encoding NMR techniques on granular media in a 3D model fluidized bed reactor. AB - A combination of PFG-NMR imaging and velocity encoding methods was applied to investigate the dynamic behavior of a bed of poppy seeds subjected to air flow, representing a model setup for fluidized bed reactors. The particle motion is described both from a statistical point of view, by determining propagators and dispersion coefficients representing an average over the whole bed volume, as well as combined with spatial resolution by generating velocity maps. Velocity images of different horizontal slices in the bed confirm the notion of a toroidal particle flow pattern inside the shallow granular bed. Despite the need of considerable averaging due to the random motion of the relatively few particles in the bed, quantitative velocity images and statistical information about the random particle motion can be obtained from monitoring the fluid component in the seeds by conventional spin-echo techniques. PMID- 16269262 TI - Differential expression of chlamydial signal transduction genes in normal and interferon gamma-induced persistent Chlamydophila pneumoniae infections. AB - Characteristic features of the persistent chlamydial developmental cycle, associated with chronic infections in both humans and animals, include the generation of non-replicative, morphologically aberrant bodies which are distinct from normal propagating reticulate bodies. Previous studies have correlated these morphological and metabolic changes with differential expression of diverse functional subsets of chlamydial genes. To further investigate these correlations, we compared mRNA expression of predicted chlamydial signal transduction genes between normal Chlamydophila pneumoniae A-03 infections in HEp 2 cells and those treated with gamma interferon (IFN-gamma) by using real-time RT PCR. Inspection of the Cp. pneumoniae genome revealed at least 39 candidate signal transduction genes, of which 30 were differentially expressed in Cp. pneumoniae mediated persistence. Functional sub-groups of differentially expressed signal transduction genes include chlamydial GTPases (hflX, ychF, yhbZ and yphC), linked to bacterial cellular processes such as cell cycle control and ribosome assembly and stability. Other up-regulated signal transduction genes sharing similarity to bacterial stress response genes (htrA, surE, lytB and hrcA) were also detected. The transcriptional changes observed for the majority of signal transduction genes appear to be unique for Cp. pneumoniae, as similar changes were not observed in recent whole genomic analysis of C. trachomatis IFN gamma mediated persistence. These results suggest that chlamydial signal transduction genes play potentially important roles in the establishment and maintenance of Cp. pneumoniae persistence, likely as part of the IFN-gamma response stimulon as described for C. trachomatis, but with considerable differences in the transcriptional profile. PMID- 16269263 TI - Monokine induced by interferon gamma and IFN-gamma response to a fusion protein of Mycobacterium tuberculosis ESAT-6 and CFP-10 in Brazilian tuberculosis patients. AB - IFN-gamma responses to Mycobacterium tuberculosis antigens ESAT-6 and CFP-10 have been proposed as specific markers of M. tuberculosis infection. Monokine induced by gamma interferon (MIG/CXCL9) has been shown to be expressed by IFN-gamma stimulated mononuclear cells and to attract activated T-cells through the chemokine receptor CXCR3. Since MIG is induced early in the response to IFN gamma, measuring MIG may provide an interesting marker to assess downstream IFN gamma induced responses, in contrast to assays that mainly focus on quantifying production of IFN-gamma per se. We, therefore, investigated MIG and IFN-gamma responses to a fusion protein of ESAT-6 and CFP-10, and compared responses to the conserved mycobacterial antigen 85B (Ag85B) and purified protein derivative (PPD) of M. tuberculosis, in 29 BCG vaccine controls and 24 TB patients. IFN-gamma secreting cells were determined by ELISPOT, and MIG production was measured by ELISA and flow cytometry. Production of MIG in response to ESAT-6/CFP-10, Ag85B and PPD correlated overall with increased numbers of IFN-gamma secreting cells (r=0.55, P<0.0001). A significant increase was noted among patients compared to controls in the secretion of IFN-gamma and MIG following stimulation with ESAT 6/CFP-10 or PPD (P<0.05). Moreover, MIG intracellular expression was higher in TB patients compared to BCG vaccines (P<0.05) in response to ESAT-6/CFP-10 or PPD. We conclude that MIG production correlates significantly with enhanced T-cell IFN gamma production induced by M. tuberculosis-specific antigens ESAT-6/CFP-10. These results point to MIG as a potential novel biomarker that may be helpful in assessing downstream responses induced by IFN-gamma in TB. PMID- 16269264 TI - Establishment of Vero E6 cell clones persistently infected with severe acute respiratory syndrome coronavirus. AB - Little information is available on persistent infection of severe acute respiratory syndrome (SARS) coronavirus (CoV). In this study, we established persistent infection of SARS-CoV in the Vero E6 cell line. Acute infection of Vero E6 with SARS-CoV produced a lytic infection with characteristic rounding cytopathic effects (CPE) and the production of a large number of infectious particles in the culture fluid within 3 days post-infection. Upon subsequent culturing of the remaining adherent cells, the cells gradually proliferated and recovered normal morphology similar to that of the parental cells, and continued to produce large numbers of infectious viral particles during the observation period of 5 months. Among a total of 87 cell clones obtained from the persistently infected Vero E6, only four cell clones (named #13, #18, #21, and #34) were positive for viral RNA. Clones #13, #18, and #34 shifted to viral RNA negative during subsequent cultures, while #21 continuously produced infectious particles at a high rate. The SARS-CoV receptor, angiotensin-converting enzyme 2, was almost completely down regulated from the cell surface of persistently infected cells. Western blot analysis as well as electron microscopy indicated that the ratios of spike to nucleocapsid protein in clone #21 as well as its parental persistently infected cells were lower than that in the cells in the acute phase of infection. These Vero E6 cells persistently infected with SARS-CoV may be useful for clarifying the mechanism of the persistent infection and also for elucidating the possible pathophysiologic significance of such long-term maintenance of this virus. PMID- 16269265 TI - TNF but not Fas ligand provides protective anti-L. major immunity in C57BL/6 mice. AB - The pro-inflammatory cytokine TNF is essential for a protective immune response to some but not all strains of Leishmania major. TNF-deficient mice of a resistant genetic background succumbed rapidly to an infection with L. major BNI. Another member of the TNF superfamily, Fas ligand (FasL), has also been reported to be critical for the immune response to L. major. To test the relative importance of TNF versus FasL for the control of L. major BNI, we infected wildtype C57BL/6 (B6.WT), B6.TNF(-/-), B6.gld and C57BL/6.gld x TNF(-/-) (B6.gld.TNF(-/-)) double-negative mice. Visceral, fatal disease was only observed in B6.TNF(-/-) mice, but not in B6 gld mice. The course of infection and the immune response of B6.gld.TNF(-/-) mice were similar to those of B6.TNF(-/-) mice. B6.gld.TNF(-/-) mice had a high tissue parasite burden and expressed prominent amounts of inducible nitric oxide synthase (iNOS) in the skin, the lymph nodes (LN) and the spleen as previously reported for B6.TNF(-/-) mice, whereas the tissue parasite load and the iNOS expression of B6.gld mice resembled that of B6.WT controls. Neither the TNF- nor the FasL-deficiency exerted a detectable intrinsic effect on the proliferation of T cells. Thus, TNF, but not FasL is essential for the control of L. major BNI. The discrepancy between these and other published data are most likely due to the use of different strains of the pathogen. PMID- 16269266 TI - Geographic and ethnic differences in frequencies of two polymorphisms (D/N394 and L/I272) of the parkin gene in sporadic Parkinson's disease. AB - In this report, we evaluated the allele frequency of the D/N394 single nucleotide polymorphism (SNP) in exon 11 of the parkin gene in 200 Japanese patients with sporadic Parkinson's disease (PD) and 200 normal controls. Although the reported allele frequency of G-to-A (D/N394) is 2% in Caucasians, this SNP was not detected in Japanese patients and healthy controls. Evaluation of L/I272 polymorphism, a C-to-A transition in exon 7, showed the polymorphism in only six controls, but not in PD patients. Our results suggest that the frequencies of parkin polymorphisms are different among Asians and Caucasians. PMID- 16269267 TI - Neuroimmunoendocrine circuitry of the 'brain-skin connection'. AB - The skin offers an ideally suited, clinically relevant model for studying the crossroads between peripheral and systemic responses to stress. A 'brain-skin connection' with local neuroimmunoendocrine circuitry underlies the pathogenesis of allergic and inflammatory skin diseases, triggered or aggravated by stress. In stressed mice, corticotropin-releasing hormone, nerve growth factor, neurotensin, substance P and mast cells are recruited hierarchically to induce neurogenic skin inflammation, which inhibits hair growth. The hair follicle is both a target and a source for immunomodulatory stress mediators, and has an equivalent of the hypothalamus-pituitary-adrenal axis. Thus, the skin and its appendages enable the study of complex neuroimmunoendocrine responses that peripheral tissues launch upon stress exposure, as a basis for identifying new targets for therapeutic stress intervention. PMID- 16269268 TI - Antioxidant enzymes in ringed seal tissues: potential protection against dive associated ischemia/reperfusion. AB - Diving seals experience heart rate reduction and preferential distribution of the oxygenated blood flow to the heart and brain, widespread peripheral vasoconstriction, and selective ischemia in the most hypoxia-tolerant tissues. The first breath after the dive restores the oxygenated blood flow to all tissues and raises the potential for the production of reactive oxygen species (ROS). We hypothesized that in order to counteract the damaging effects of ROS and to tolerate repetitive cycles of ischemia/reperfusion associated with diving, ringed seal (Phoca hispida) tissues have elevated activities of antioxidant enzymes. Activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx) and glutathione-S-transferase (GST) were measured by spectrophotometric techniques in heart, kidney, liver, lung, and muscle extracts of ringed seals and domestic pigs (Sus scrofa). The results suggest that in ringed seal heart SOD, GPx and GST activities are an efficient protective mechanism for counteracting ROS production and its deleterious effects. Apparently CAT activity in seal liver and GPx activity in seal muscle participate in the removal of hydroperoxides, while seal lung appears to be protected from oxidative damage by SOD and GPx activities. PMID- 16269269 TI - Janus kinase-2 signaling mediates apoptosis in rat cardiomyocytes. AB - We tested the hypothesis that activation Jak2, which is prominently involved in the up-regulation of the renin-angiotensin system (RAS), constitutes a focal point in relaying signals triggered by a Angiotensin II (Ang II) and hypoxia/reoxygenation separately to cause an enhanced susceptibility of cardiac myocyte to apoptotic cell death. Ang II-treated adult cardiomyocytes in culture exhibited an increased level of apoptosis that accompanied activation of pro apoptotic as well as anti-apoptotic signaling pathways. We observed increased phosphorylation of Jak2 kinase, Stat1, JNK, with increased expression of Bax protein, followed by an increase in caspase-1 and caspase-3 activity. Activation of these pro-apoptotic pathways was blocked by the Jak2 pharmacological inhibitor, Tyrphostin AG490. We also observed an increase in phosphorylation of cardioprotective pathway components, namely S6 ribosomal protein, and heat shock protein 27 (HSP27). Likewise, the oxidative stress, via the hypoxia/reoxygenation treatment of rat adult cardiomyocytes, produced apoptosis that was dependent upon activation of Jak2. The apoptotic response was not only reduced by Losartan, an inverse agonist of the AT1, receptor, but by treatment with AG490 as well. Taken together, these observations provide clear evidence in favor of Jak2 signaling as mediator of the apoptotic response in cardiomyocytes. However, there was a concomitant induction of cytoprotective signaling that presumably provides a negative feed-back to the deleterious effects of the agonist. PMID- 16269271 TI - A review of developmental instability, parasitism and disease. Infection, genetics and evolution. AB - Developmental instability is reflected in imprecise development caused by perturbations of the developmental process, while developmental stability reflects the ability to avoid or reduce such perturbations by developmental means. Developmental instability has been hypothesized to reflect overall individual condition, and asymmetric or otherwise aberrant individuals have thus been predicted to be particularly severely affected by disease and parasitism. An extensive review of the literature on animals, including humans, revealed consistent relationships between increased bilateral asymmetry and elevated risk of parasitism. Parasitism, including parasitism of mothers, is a cause of asymmetry as shown by a number of experiments, and asymmetric individuals are differentially susceptible to a range of different parasites. Extensive studies of humans have shown that asymmetric individuals also suffer disproportionately from a range of different diseases including mental diseases. Studies of transgenic organisms have now demonstrated that single genes associated with disease is a sufficient cause of increased asymmetry. A number of studies have also shown that activation of the immune system causes increased asymmetry in developing individuals, and that asymmetry and immunity show negative covariation. These findings may have important implications for the study of susceptibility of hosts to infectious disease. PMID- 16269270 TI - Antithrombotic effect of the type III collagen-related octapeptide (KOGEOGPK) in the mouse. AB - Platelet adhesion to subendothelial types I and III collagens exposed upon vascular injury plays a crucial role in hemostasis and thrombosis. We previously identified a KOGEOGPK sequence (O for hydroxyproline) within type III collagen interacting with platelets, and demonstrated a strong inhibitory effect of the KOGEOGPK peptide on human platelet interactions with type III collagen in vitro. In the present study, we tested the antithrombotic effect of KOGEOGPK in vivo. In a mouse model of pulmonary thromboembolism induced by intravenous injection of type III collagen and epinephrine, prior administration of 80 mg/kg KOGEOGPK reduced by 50% the size of thrombi embolized in lungs, compared to vehicle treated mice (p<0.0001). In a mouse model of photochemically induced lesion of caecum venules and arterioles, intravenous injection of 80 mg/kg KOGEOGPK decreased by 76% the occurrence of arteriole occlusion 45 min after vascular injury (p<0.05). A moderate antithrombotic effect of KOGEOGPK was also observed in the injured venules. In addition, intracardiac injection of KOGEOGPK had no effect on the tail bleeding time. These findings demonstrate a substantial contribution of platelet interactions with the type III collagen-related KOGEOGPK sequence in thrombus formation in vivo with preferential involvement in arterial thrombosis. PMID- 16269272 TI - Ultrasound assisted extraction in quantifying lutein from chicken liver using high-performance liquid chromatography. AB - Four sample preparation methods, (1) solvent (SOL), (2) saponification and solvent (SP), (3) ultrasound assisted solvent (UA), and (4) saponification and ultrasound assisted solvent (SP-UA), were used for quantifying lutein in chicken liver samples by HPLC. The lutein concentrations obtained by using SOL, UA, SP, and SP-UA were significantly different with values from 10.4 microg/g (UA) to undetected (SOL). Efficiency of the four different methods for extracting lutein from high to low were the UA, SP, SP-UA, and SOL method. The measured value of lutein in the liver sample using the UA method was approximately two and three times higher than that obtained from the SP and SP-UA method, respectively. The methods with saponification significantly affected the stabilities of lutein in liver samples. The lutein concentration measured with the solvent only method was either much lower than any of the other extraction methods or undetectable. This indicated that little lutein in those samples was in a form that could be extracted directly by solvent. Compared with the saponification method, the ultrasound assisted solvent method could effectively extract lutein from sample matrix and thus avoid chemical degradation reactions, which would be especially important for complex biological tissue such as liver. PMID- 16269273 TI - Sensitive liquid chromatography-tandem mass spectrometry method for the determination of scutellarin in human plasma: Application to a pharmacokinetic study. AB - A sensitive and selective liquid chromatographic-tandem mass spectrometric (LC MS/MS) method for the determination of scutellarin in human plasma has been developed. Samples were prepared using solid phase extraction and analyzed on a C(18) column interfaced with a triple quadrupole tandem mass spectrometer. Positive electrospray ionization was employed as the ionization source. The mobile phase consisted of methanol-water (0.1% formic acid), using gradient procedure. The analyte and internal standard baicalin were both detected by use of selected reaction monitoring mode. The method was linear in the concentration range of 0.2-20.0 ng/mL. The lower limit of quantification (LLOQ) was 0.2 ng/mL. The intra- and inter-day relative standard deviation across three validation runs over the entire concentration range was less than 12.4%. The accuracy determined at three concentrations (1.0, 5.0 and 10.0 ng/mL for scutellarin) was within +/ 5.0% in terms of relative error. The method herein described was successfully applied for the evaluation of pharmacokinetic profiles of scutellarin guttate pills in 20 healthy volunteers. PMID- 16269274 TI - SPE-GC-MS for the sampling and determination of unmetabolized styrene in urine. AB - The urinary excretion of unmetabolized styrene can be a very good indicator for biomonitoring styrene in occupationally exposed people. The use of a new urine sampling system, involving a solid-phase extraction cartridge, offers several advantages for determining styrene. The advantages are especially related to the pre-analytical phase of styrene determination, which may be influenced by many variables. The effect on styrene recovery of sorbent type, eluting solvent, elution volume, elution flow-rate, and the addition of methanol to the washing solvent, was evaluated by experimental design methodology. As a result, Oasis HLB cartridges were selected for urine sampling, as well as 1.5 mL of ethyl acetate at 0.5 mL/min for eluting the retained styrene. These conditions were then applied to the validation of the solid-phase extraction combined with GC-MS method for the sampling and analysis of unmetabolized styrene in urine. The overall uncertainty was in the 12-22% range and the limit of detection was 2.2 microg/L for a 4 mL urine sample. The stability of styrene has been studied both in cartridges and in vials under different storage periods. After 1 month period the styrene stored on cartridges at room temperature remained stable, whereas this is not the case for styrene recovery from vials. The results obtained indicate that on-site solid-phase extraction of urine can provide a simple, accurate and reproducible sampling and analytical method for the biomonitoring of styrene in urine. PMID- 16269275 TI - Identification and characterization of digestive serine proteases from inhibitor resistant Helicoverpa zea larval midgut. AB - Protease inhibitors mediate a natural form of plant defence against insects, by interfering with the digestive system of the insect. In this paper, affinity chromatography was used to isolate trypsins and chymotrypsins from Helicoverpa zea larvae, which had been raised on inhibitor-containing diet. Sensitivity of the fractions to inhibition by plant proteinase inhibitors was tested, and compared to the sensitivity of proteinases found in insects raised on diet to which no inhibitor had been added. The isolated chymotrypsin activity was found to be less sensitive to plant protease inhibitors. The sensitivity of the isolated trypsin activity was found to be intermediate between completely sensitive trypsins and completely insensitive forms that have been previously described. Mass spectrometry was used to identify one trypsin and two chymotrypsins in the partially purified protease fraction. The sequence features of these proteases are discussed in relation to their sensitivity to inhibitors. The results provide insight in the enzymes deployed by Helicoverpa larvae to overcome plant defence. PMID- 16269276 TI - Sensitive assay for determining plasma tenofovir concentrations by LC/MS/MS. AB - An LC/MS/MS assay for the determination of tenofovir (TNF) was developed and validated for use with the EDTA anticoagulated human plasma matrix. Heparin treated plasma and serum matrices were also validated. After addition of adefovir as an internal standard, trifluoroacetic acid was used to produce a protein-free extract. Chromatographic separation was achieved with a Polar-RP Synergi, 2.0 mm x 150 mm, reversed-phase analytical column. The mobile phase was 3% acetonitrile/1% acetic acid, aq. Detection of TNF and the internal standard was achieved by ESI MS/MS in the positive ion mode using 288/176 and 274/162 transitions, respectively. The method was linear from 10 to 750 ng/ml with a minimum quantifiable limit of 10 ng/ml when 250 microl aliquots were analyzed. The usefulness of this LC/MS/MS method to routinely monitor plasma concentrations of TNF was demonstrated along with its ability to assist in the performance of pharmacokinetic studies. PMID- 16269277 TI - Microdialysis sampling coupled to on-line high-performance liquid chromatography for determination of arbutin in whitening cosmetics. AB - We have developed a method that uses on-line microdialysis sampling coupled with high-performance liquid chromatography (HPLC) to determine arbutin in whitening cosmetics. The optimum analytical conditions for microdialysis sampling were a probe length of 10mm and a dialysis flow-rate of 5 microl min-1. The accuracy (% bias) for intra-day (n=6) and inter-day (n=30, five consecutive days) analyses ranged from -8.9 to 11.5%, with a precision below 7.64% R.S.D. The calibration curve was linear within the range from 0.1 to 20 mM (R2=0.9989). The detection limit was 15 microM. By comparing the arbutin levels determined this way in the whitening cosmetics with the results obtained from the no-net-flux method, we conclude that our proposed on-line microdialysis-HPLC system displays good accuracy. We evaluated the robustness of our optimum conditions by means of a Plackett-Burman design. Apart from the effect of a low flow-rate of perfusate--an increase of 12.52+/-2.31%--we observed no significant changes in the analyses upon changing the levels of any other parameter. Because this on-line method offers the advantages of simplicity, reliability, the lack of any tedious sample pretreatment process, and a reduced use of organic solvents, we believe that it is suitable for the routine analysis of commercial cosmetics. PMID- 16269278 TI - Biochemical characterization of a truncated penta-EF-hand Ca2+ binding protein from maize. AB - Plants possess multiple genes encoding calcium sensor proteins that are members of the penta-EF-hand (PEF) family. Characterized PEF proteins such as ALG-2 (apoptosis-linked gene 2 product) and the calpain small subunit function in diverse cellular processes in a calcium-dependent manner by interacting with their target proteins at either their N-terminal extension or Ca2+ binding domains. We have identified a previously unreported class of PEF proteins in plants that are notable because they do not possess the hydrophobic amino acid rich N-terminal extension that is typical of these PEF proteins. We demonstrate that the maize PEF protein without the N-terminal extension has the characteristics of known PEF proteins; the protein binds calcium in the 100 nM range and, as a result of calcium binding, displays an increase in hydrophobicity. Characterization of the truncated maize PEF protein provides insights into the role of the N-terminal extension in PEF protein signaling. In the context of the current model of how PEF proteins are activated by calcium binding, these results demonstrate that this distinctive class of PEF proteins could function as calcium sensor proteins in plants even in the absence of the N terminal extension. PMID- 16269279 TI - Crystallography for protein kinase drug design: PKA and SRC case studies. AB - Protein crystallography can be used throughout the drug discovery process to obtain diverse information critical for structure based drug design. At a minimum, a single target structure may be available. Optimally, and especially for protein kinases, a broad range of crystal structures should be obtained to characterize target flexibility, structure modulation via co-factor binding or posttranslational modification, ligand induced conformational changes, and off target complex structures for selectivity optimization. The flexibility of the protein kinases is in contrast to the need for "crystallizable" constructs, that is, proteins that crystallize under varying conditions and in varying crystal packing arrangements. Strategies to produce crystallizable protein kinase constructs include truncation to the catalytic domain, co-crystallization with rigidifying ligands, crystallization of known rigid forms, and point mutation to improve homogeneity or mimic less crystallizable proteins. PKA, the prototypical serine/threonine protein kinase, and SRC, a tyrosine kinase and the first identified oncoprotein, provide multiple examples of these various approaches to protein kinase crystallography for drug design. PMID- 16269280 TI - The NK-lysin derived peptide NK-2 preferentially kills cancer cells with increased surface levels of negatively charged phosphatidylserine. AB - The NK-lysin derived peptide NK-2 is a potent antibacterial, but non-toxic to a human keratinocyte cell line and of low hemolytic activity. Its target selectivity is based upon a strong binding preference to membranes containing anionic phospholipids, which are normally not found on the surface of human cells. Here, we analyzed the interaction of NK-2 with normal human lymphocytes and seven different human cancer cell lines and demonstrate that some of these cells expose negatively charged surface phosphatidylserine (PS), which presumably facilitates killing of the cells by NK-2. This is underlined by the specific intercalation of the peptide into PS-containing liposomes analyzed by fluorescence-resonance energy transfer spectroscopy. PMID- 16269281 TI - President's address: future challenges. PMID- 16269282 TI - Computer-enhanced robotic telesurgery minimizes esophageal perforation during Heller myotomy. AB - BACKGROUND: Laparoscopic Heller myotomy has emerged as the treatment of choice for achalasia. However, intraoperative esophageal perforation remains a significant complication. Computer-enhanced operative techniques have the potential to improve outcomes for certain operative procedures. Robotic, computer enhanced laparoscopic telemanipulators using 3-dimensional magnified imaging and motion scaling are designed uniquely to facilitate certain operations requiring fine-tissue manipulation. We hypothesized that computer-enhanced robotic Heller myotomy would reduce intraoperative complications compared with laparoscopic techniques. METHODS: All patients undergoing an operation for achalasia at 3 institutions with a robotic surgery system (DaVinci; Intuitive Surgical Corporation, Sunnyvale, Calif) were followed-up prospectively. Demographics, perioperative course, complications, and hospital stay were recorded. Follow-up evaluation was obtained via a standardized symptom survey, office visits, and medical records. Data were compared with preoperative symptoms using a Mann Whitney U test, and operating times were compared using the ANOVA test. RESULTS: Between August 2000 and August 2004 there were 104 patients who underwent a robotic Heller myotomy with partial fundoplicaton. There were 53 women and 51 men. All patients were symptomatic. The operative time was 140.55 minutes overall, but improved from 162.63 minutes to 113.50 minutes from 2000-2002 to 2003-2004 (P = .0001). There were no esophageal perforations. There were 8 minor complications and 1 patient required conversion to an open operation. Sixty-six (62.3%) patients were discharged on the first postoperative day and the average hospital stay was 1.5 days. A symptom survey was completed in 79 of 104 patients (76%) at follow-up evaluation. Symptoms improved in all patients with an average follow-up symptom score of 0.48 compared with 5.0 before the operation (P = .0001). Forty-three of the 79 patients from whom follow-up data were collected had a minimum follow-up period of 1 year. The follow-up period averaged 16 months. No patients required reoperation. CONCLUSIONS: Computer-enhanced robotic laparoscopic techniques provide a clear advantage over standard laparoscopy for the operative treatment of achalasia. We have shown in this large series that Heller myotomy can be completed using this technology without esophageal perforation. The application of computer-enhanced operative techniques appears to provide superior outcomes in selected procedures. PMID- 16269283 TI - Surgical considerations in cystic fibrosis: a 32-year evaluation of outcomes. AB - BACKGROUND: Information concerning long-term operative outcomes in patients with cystic fibrosis (CF) is relatively sparse in the operative literature. METHODS: A retrospective review of CF patients with operative conditions was performed (1972 2004) at a tertiary children's hospital to analyze outcomes including long-term morbidity and survival. RESULTS: A total of 226 patients with CF presented with an operative diagnosis (113 men, 113 women). A total of 422 operations were performed in 213 patients (94%). The mean age at operation was 4.1 +/- 6.2 years (range, 1 d to 26 y) and 109 were neonates. Fifteen of 42 (36%) babies with simple meconium ileus (MI) were treated nonoperatively with hypertonic enemas, 27 of 42 and all 45 patients with complicated MI required operation, including 15 with jejunoileal atresia (17%). Seventeen of 27 (63%) patients with meconium ileus equivalent had MI as neonates; 7 of 27 (26%) required operation. Eight of 9 (89%) with fibrosing colonopathy required operation. Organ transplantation was required in 21 patients. Follow-up evaluation was possible in 204 of 213 (96%) patients. The duration of follow-up evaluation was 14.9 +/- 8.5 years (range, 2 mo to 35 y). Operative morbidity was 11% at 1 year, 2% at 2 to 4 years, 1% at 5 to 10 years, and less than 1% at more than 10 years. There were 24 deaths (11%); 22 followed CF-related pulmonary complications and included 8 of 16 (50%) children with pneumothorax. CONCLUSIONS: Long-term survival in CF patients has improved significantly (89%), with many surviving into the fourth decade. MI may predispose to late complications including meconium ileus equivalent and fibrosing colonopathy. Pneumothorax in CF patients is an ominous predictor of mortality. Children with CF are living longer and are good candidates for operation, but require long-term follow-up evaluation because of ongoing exocrine dysfunction. PMID- 16269284 TI - Molecular epidemiology of microbial contamination in the operating room environment: Is there a risk for infection? AB - BACKGROUND: Modern operating rooms are considered to be aseptic environments. The use of surgical mask, frequent air exchanges, and architectural barriers are used to reduce airborne microbial populations. Breaks in surgical technique, host contamination, or hematogenous seeding are suggested as causal factors in these infections. This study implicates contamination of the operating room air as an additional etiology of infection. METHODS: To investigate the potential sources of perioperative contamination, an innovative in situ air-sampling analysis was conducted during an 18-month period involving 70 separate vascular surgical procedures. Air-sample cultures were obtained from multiple points within the operating room, ranging from 0.5 to 4 m from the surgical wound. Selected microbial clonality was determined by pulse-field gel electrophoresis. In a separate series of studies microbial nasopharyngeal shedding was evaluated under controlled environmental conditions in the presence and absence of a surgical mask. RESULTS: Coagulase-negative staphylococci were recovered from 86% of air samples, 51% from within 0.5 m of the surgical wound, whereas Staphylococcus aureus was recovered from 64% of air samples, 39% within 0.5 m from the wound. Anterior nares swabs were obtained from 11 members of the vascular team, clonality was observed between 8 strains of S epidermidis, and 2 strains of S aureus were recovered from selected team members and air-samples collected throughout the operating room environment. Miscellaneous Gram-negative isolates were recovered less frequently (<33%); however, 7 isolates expressed multiple patterns of antimicrobial resistance. The traditional surgical mask demonstrated limited effectiveness at curtailing microbial shedding, especially during symptomatic periods of rhinorrhea. CONCLUSIONS: Gram-positive staphylococcal isolates were frequently isolated from air samples obtained throughout the operating room, including areas adjacent to the operative field. Nasopharyngeal shedding from person participating in the operation was identified as the source of many of these airborne contaminants. Failure of the traditional surgical mask to prevent microbial shedding is likely associated with an increased risk of perioperative contamination of biomedical implants, especially in procedures lasting longer than 90 minutes. PMID- 16269285 TI - Intraoperative parathyroid hormone testing improves cure rates in patients undergoing minimally invasive parathyroidectomy. AB - BACKGROUND: Intraoperative parathyroid hormone (iPTH) testing often is used during minimally invasive parathyroidectomy for primary hyperparathyroidism (1 degrees HPT). However, several investigators report that these assays are not cost effective and do not improve outcomes significantly. METHODS: To determine the impact of iPTH testing on the outcomes of patients with 1 degrees HPT, we reviewed our experience. From January 1990 to June 2004, there were 345 consecutive patients with 1 degrees HPT and positive localization studies for a single parathyroid adenoma who were candidates for minimally invasive parathyroidectomy. Group 1 patients (n = 157) underwent parathyroid exploration without iPTH testing and group 2 patients (n = 188) had an operation with iPTH testing. RESULTS: Of the group 1 patients, 15 (10%) still were hypercalcemic postoperatively owing to additional unidentified hyperfunctioning parathyroid glands. In contrast, among 188 group 2 patients, 170 (90%) had resection of a single parathyroid adenoma, a greater than 50% decrease in iPTH levels, and were cured. The remaining 18 (10%) patients did not have an adequate reduction in iPTH levels and underwent bilateral neck exploration with resection of additional parathyroids. Of these 18 patients, 9 had double adenomas and 9 had 3- or 4-gland hyperplasia. Importantly, all patients in group 2 were cured. CONCLUSIONS: iPTH testing improves cure rates in patients undergoing minimally invasive parathyroidectomy. iPTH testing allowed intraoperative recognition and resection of additional hyperfunctioning parathyroids missed by preoperative imaging studies. Consequently, we strongly advocate the routine use of iPTH testing in patients who undergo minimally invasive parathyroidectomy for 1 degrees HPT. PMID- 16269286 TI - Comparative analysis of minimally invasive microductectomy versus major duct excision in patients with pathologic nipple discharge. AB - BACKGROUND: Minimally invasive techniques are being used increasingly in patients with benign and malignant breast diseases. The purpose of this study was to compare the diagnostic yield of 2 groups of patients who underwent either minimally invasive microductectomy or major duct excision for pathologic nipple discharge. METHODS: Two hundred thirty-five patients who underwent nipple exploration and duct excision and were part of an institutional review board approved database were included in this retrospective analysis. Preoperative imaging, ductal washing cytology, surgical specimen size, and final histopathology were compared among 95 patients who underwent microductectomy by using intraoperative ductoscopy and 140 patients undergoing standard major duct excision. RESULTS: Mean age of patients undergoing microductectomy was 53 versus 55 years in patients undergoing major duct excision. Preoperative mammogram was negative or benign in 92% and suspicious in 8% of patients in both the microductectomy group and the major duct excision group. A ductal abnormality was identified by preoperative ductography in 43 of 56 (77%) patients in the microductectomy group versus 74 of 92 (80%) patients in the major duct excision group. Ductal cytology was benign in 81% and 80% of patients tested, respectively. Mean specimen size was significantly smaller in patients who underwent microductectomy (9.2 cm3) as compared with major duct excision (12.6 cm3). Although the percentage of patients with atypical ductal hyperplasia or lobular carcinoma in situ was similar among the 2 groups (9% vs 10%), only 3 of 95 (3%) patients within the microductectomy group were found to have carcinoma within the resection specimen as compared with 12 of 140 (9%) within the major duct excision group (P = .03). Mean specimen size of the patients diagnosed with carcinoma was 8.6 cm(3) in the microductectomy group as compared with 15.5 cm3 in the major duct excision group (P = .014). CONCLUSIONS: These data confirm that patients who present with single duct pathologic nipple discharge usually have benign pathology as the etiology. However, in a small percentage of patients an occult carcinoma might be present. Major duct excision appears to detect a higher percentage of occult carcinoma when compared with minimally invasive microductectomy, which might be related to the larger sample size of the resection specimen. PMID- 16269287 TI - Endovascular repair of thoracic aortic pathology with custom-made devices. AB - BACKGROUND: Open repair of thoracic aortic aneurysms (TAAs) is fraught with high morbidity and mortality rates. The availability of endoprostheses for treating thoracic aortic pathology has not kept pace with those for treating abdominal aneurysms. Technical feasibility, durability, and safety of custom-made stent grafts for the treatment of TAAs and dissections are evaluated. METHODS: From July 2002 to October 2004 there were 15 patients with TAAs, intramural hematoma, or dissections treated with custom-made endografts. Grafts were deployed after brief adenosine-induced cardiac arrest. Computed tomography scans were obtained 1 month postoperatively and every 6 months thereafter. The mean follow-up period was 15 months (range, 3-31 mo). RESULTS: The mean age of patients was 67 +/- 11 years (range, 47-81 y; 67% men, 33% women). Indications for repair included TAA (10), chronic type B dissection (3), penetrating ulcer (1), and acute dissection (1). Planned concomitant procedures included subclavian-carotid transposition (2) and aortosplenic bypass (2) to achieve adequate proximal or distal landing seal zones, respectively. The mean length of hospital stay was 8 days (range, 1-49 d). Immediate complications included 2 access-related events, 1 cerebellar infarction treated expectantly, and 1 death from a large hemispheric stroke. There were no cases of postoperative paralysis and on follow-up imaging no cases of endoleak, endograft migration, or stent fractures were found. No late deaths occurred caused by stent-graft repair or aneurysm-related causes. CONCLUSIONS: Endoluminal exclusion of thoracic aortic aneurysms and dissections can be achieved successfully using custom-made stent-grafts. The use of specially designed devices appears to be technically feasible and durable, with acceptable morbidity and mortality rates. PMID- 16269288 TI - Selective management of blunt hepatic injuries including nonoperative management is a safe and effective strategy. AB - BACKGROUND: The justification and preference for operative versus nonoperative management of hepatic injuries caused by blunt trauma remains ambiguous. This review assesses the outcome of operative and nonoperative management of liver injury after blunt trauma. METHODS: We retrospectively reviewed the demographics, severity of injury, severity of liver injury, associated concomitant injuries, management scheme, and outcome of patients with documented hepatic injury from 1993 to 2003. RESULTS: The overall mortality rate was 9.4%, with 3.7% caused by the liver injury itself. Fifty-nine percent (330 of 561) of liver injuries were of low severity (grades I and II), with an overall mortality rate of 6.6% caused by concomitant injuries and liver-related mortality of 0%. Forty-one percent (231 of 561) of liver injuries were high-severity injuries (grades III, IV, and V). Mortality for nonoperative management of high-severity liver injuries was 2.2%. If operative intervention was required because of hemodynamic instability or concomitant injuries then the mortality rate was significantly higher at 30%. Forty-two of the 378 (11%) liver injuries treated nonoperatively required an adjunctive procedure for successful management. CONCLUSIONS: Selective management of liver injuries presented a low liver-related mortality rate. Low-grade injuries can be managed nonoperatively with excellent results. High-grade injuries can be managed nonoperatively, if operative intervention is not required for hemodynamic instability or associated injuries, with a low mortality. In these patients, adjunctive procedures will be required selectively for successful nonoperative management of high-grade liver injuries. High-grade injuries requiring operative management because of hemodynamic instability or concomitant injuries continue to have significantly higher mortality. PMID- 16269289 TI - Efficacy of laparoscopic fundoplication in controlling pulmonary symptoms associated with gastroesophageal reflux disease. AB - BACKGROUND: Gastroesophageal reflux disease (GERD)-induced pulmonary symptoms (PS) can be difficult to control. The effectiveness of laparoscopic fundoplication (LF) in controlling PS among patients with medically recalcitrant GERD is poorly documented. We evaluated our results in controlling important PS in patients with GERD undergoing LF. METHODS: Seventy-four patients (28 men, 46 women) were identified with clinically important PS from a prospective cohort of 155 patients undergoing elective LF for recalcitrant GERD. Median age was 52.5 years (range, 29-84 years). Sixty-seven (91%) patients were taking proton pump inhibitors at the time of operation. Quality of life by using the SF36 physical (PCS) and mental (MCS) component summary scores (normal, 50) and heartburn severity by using the health-related quality of life (HRQOL) (best score, 0; worst score, 45) were measured. RESULTS: All 74 patients with PS survived operation, and minor morbidity occurred in 5 (7%) patients. Median hospital stay was 2 days (range, 1-6 days), and return to normal activity was seen at 2.2 weeks (range, 1-8 weeks). Median follow-up was 12 months. PS were improved significantly (P < .01) for hoarseness (62% to 17.6%), bronchospasm (60% to 9.5%), and aspiration (22% to 1.4%). Before LF, 11 (14.9%) patients required bronchodilators or oral steroids. Postoperatively such therapy was required in only 3 (4.2%) patients (P = .019), with no patient requiring oral steroids. Patients with poorer control of their GERD on the basis of high HRQOL scores had significantly more PS after operation. CONCLUSIONS: A significant number of patients with medically recalcitrant GERD (46% from our prospective database) have important PS. LF can improve PS, decrease requirement for pulmonary medications, as well as improve typical reflux symptoms and quality of life. PMID- 16269290 TI - A prospective randomized trial comparing standard pancreatoduodenectomy with pancreatoduodenectomy with extended lymphadenectomy in resectable pancreatic head adenocarcinoma. AB - BACKGROUND: To compare operative morbidity, mortality, quality of life, and survival after pancreatoduodenectomy (PD) versus pancreatoduodenectomy with extended lymphadenectomy (PD/ELND) in patients with resectable pancreatic cancer. METHODS: From May 1997 to July 2003 there were 132 patients with biopsy examination-proven or suspected adenocarcinoma of the pancreatic head who agreed to participate in a single-institution, prospective, randomized trial. If resectable at operation, patients then were randomized to standard PD (40 patients) or PD/ELND (39 patients). Quality of life was assessed by using the Functional Assessment of Response to Cancer Therapy specific to the pancreas. Morbidity, mortality, and survival were analyzed. RESULTS: Demographics and pathologic characteristics for both groups were similar. When comparing PD/ELND with standard PD, the median operating time was greater for the PD/ELND group (7.6 h vs 6.2 h, P < .01), blood transfusion more likely (44% vs 22%, P < .05), and the median number of lymph nodes resected was greater (36 vs 15 nodes, P < .01). Morbidity and mortality rates were comparable. Median durations of stay were 11 and 10.5 days (P = NS), respectively. There were no significant differences in 1-year (71% vs 82%), 3-year (25% vs 41%), 5-year (16.5% vs 16.4%), and median (19 vs 26 mo) survival (P = .32). At 4 months postoperatively, diarrhea, body appearance, and bowel control scored lower on the Functional Assessment of Response to Cancer Therapy specific to the pancreas after PD/ELND (P < .05). CONCLUSIONS: Although a much larger study would have more power to compare statistically the survival between groups, both the decrement in quality of life and similar studies showing no survival difference make PD/ELND unattractive for further prospective investigation. PMID- 16269292 TI - Feasibility, reliability and validity of an operative performance rating system for evaluating surgery residents. AB - BACKGROUND: Resident evaluation traditionally involves global assessments including clinical performance, professional behavior, technical skill, and number of procedures performed. These evaluations lack objective assessment of operative skills. We describe an operative performance rating system (OPRS) designed to provide objective operative performance ratings using a sentinel procedure format. METHODS: Ten-item procedure-specific rating instruments were developed. Items included technical skills, operative decision making, and general items. A 1 to 5 (5 = excellent) scale was used for evaluation. Six procedures had sufficient forms returned to allow evaluation. Inter-rater reliability was determined by having faculty evaluators view 2 videotaped operations. RESULTS: Return rates for the Internet-based form were full-time faculty (92%), volunteer faculty (27%), and overall (67%). Reliability, (average interitem correlation), and total procedures evaluated were excisional biopsy, 0.90, (0.48), 77; open inguinal herniorraphy, 0.94, (0.62), 51; laparoscopic cholecystectomy, 0.95, (0.64), 75; small-bowel and colon resection, 0.92, (0.58), 30; parathyroidectomy, 0.70, (0.19), 30; and lumpectomy, 0.92, (0.51), 38. Years of training accounted for 25% to 57% of the variation in scores. Inter-rater variability was observed; however, the average rater agreement was reliable. CONCLUSIONS: Internet-based management made obtaining the data feasible. The OPRS complements traditional evaluations by providing objective assessment of operative decision-making and technical skills. Interitem correlations indicate the average rating of items provides a reliable indicator of resident performance. The OPRS is useful in tracking resident development throughout postgraduate training and offers a structured means of certifying operative skills. PMID- 16269291 TI - Surgical outcome in patients with primary sclerosing cholangitis undergoing ileal pouch-anal anastomosis: a case-control study. AB - BACKGROUND: The outcome of restorative proctocolectomy in the setting of chronic ulcerative colitis complicated by primary sclerosing cholangitis (PSC) is not clear. The purpose of this study was to determine the surgical outcome, risk of dysplasia/cancer, morbidity/mortality, long-term results, and functional and quality of life results in patients with inflammatory bowel disease (IBD) and PSC who underwent restorative proctocolectomy with ileal pouch-anal anastomosis and compare them in a case-matched study. METHODS: Patients with PSC-associated IBD undergoing restorative proctocolectomy between 1983 and 2002 were included in the study. This study group was matched for age, gender, diagnosis, duration of disease, anastomosis technique, and proximal diversion to a cohort of IBD patients with no associated PSC who underwent restorative proctocolectomy during the same period of time. Postoperative morbidity, incidence of neoplasia/cancer in the resected specimen, pouchitis, pouch failure, long-term mortality, and 5 year survival rates were compared between the groups. The functional and quality of life records were prospectively collected and compared between the groups. For each group, matched Kaplan-Meier survival analysis was also conducted comparing 5 year survival between the 2 cohorts, matching for diagnosis, duration of disease, age, gender, anastomosis type, and proximal diversion. RESULTS: Sixty-five patients with PSC and IBD underwent restorative proctocolectomy with ileal pouch anal anastomosis during the study period. Two hundred sixty IBD patients with no associated PSC who matched with the outlined criteria comprised the control group. The follow-up period was 68 +/- 50 months for the PSC group and 102 +/- 62 months for the control group. A higher incidence of cancer (14% vs 5%, P = .02) and dysplasia in the resected specimen (40% vs 7%, P < .001), an associated increased risk of postoperative pelvic sepsis (14% vs 5%, P = .02), and higher long-term mortality (35% vs 4%, P < .001) were found in the PSC group compared with control group with no associated PSC. The majority, 13 of 23 (57%), of the deaths in the PSC group were a result of liver disease. Five-year survival for the PSC group was significantly poorer than the 5-year survival for the control group with no associated PSC. No significant differences were found in functional and quality of life results between the groups in the short- and long-term follow up periods. CONCLUSIONS: PSC-associated IBD patients after restorative proctocolectomy have a higher risk of neoplasia/cancer in the resected specimen, postoperative pelvic sepsis, and higher long-term mortality. Functional and quality of life remains similar in IBD patients after restorative proctocolectomy with or without associated PSC in the follow-up. However, patients with IBD and PSC have a significantly poorer survival than patients with no associated PSC after restorative proctocolectomy. PMID- 16269293 TI - Fine-needle aspiration biopsy specimen with a predominance of Hurthle cells: a dilemma in the management of nodular thyroid disease. AB - BACKGROUND: A fine-needle aspiration biopsy (FNAB) specimen of a thyroid nodule with a predominance of Hurthle cells usually is indicative of a Hurthle cell neoplasm, but it also may occur with nonneoplastic disease. METHODS: A prospective nodular thyroid disease database was used to identify patients with a FNAB specimen consisting of a predominance of Hurthle cells. Clinical factors were investigated and FNAB specimens were examined in a blinded fashion by a single cytopathologist to determine if there were specific factors that could be used to distinguish nonneoplastic from neoplastic disease. RESULTS: Of the 738 patients with nodular thyroid disease, 622 had a FNAB specimen. The FNAB specimen was interpreted as consistent with a Hurthle cell neoplasm in 45 (7%) patients, 7 (16%) with carcinoma, 21 (47%) with adenoma, 12 (27%) with adenomatous hyperplasia, and 5 (11%) with thyroiditis. Extensive cellularity and absent colloid were associated with neoplastic disease (P < .05). No cytologic feature reliably excluded neoplastic disease (P > .05). No significant differences in age (x +/- SD) (51 +/- 17 vs 54 +/- 17 y), sex (female/male ratio, 6/1 vs 15/2), nodule size (3.9 +/- 1.9 vs 3.4 +/- 2.0 cm), weight of excised thyroid tissue (42 +/- 27 vs 33 +/- 30 g), or functional status of the thyroid gland was observed between patients with neoplastic (n = 28, 62%) versus nonneoplastic (n = 17, 38%) disease. CONCLUSIONS: Neoplastic disease accounts for two thirds of the pathology in patients with a predominance of Hurthle cells on FNAB specimen and neither clinical nor cytologic features reliably exclude Hurthle cell adenoma or carcinoma. As a result, thyroidectomy is recommended for all patients with a thyroid nodule and a predominance of Hurthle cells on FNAB specimen. PMID- 16269294 TI - Presence of distal intramural spread after preoperative combined-modality therapy for adenocarcinoma of the rectum: what is now the appropriate distal resection margin? AB - BACKGROUND: Preoperative combined-modality therapy (CMT) is considered the treatment approach of choice for rectal cancer. To examine the effect of CMT on distal intramural spread (DIS), we determined the extent of DIS in rectal cancer specimens after CMT. METHODS: Specimens from 20 patients after preoperative CMT and total mesorectal excision for rectal cancer were analyzed for extent of DIS. Specimens were washed, fixed in formalin, and sectioned in 5-mm cuts from the distal border of the residual tumor to the distal resection margin. RESULTS: Eleven patients (55%) had DIS. Two patients had a complete pathologic response (10%). The mean extent of DIS was 0.50 cm (standard deviation, 0.64 cm; range, 0.0 to 2.5 cm). Only 1 patient in the study had DIS beyond 1 cm from the residual tumor. Six of the patients have died at a median of 30.4 months after operation (range, 19.0 to 51.1 months). No correlation was found between the extent of residual DIS and survival. CONCLUSIONS: Our study shows that the vast majority of patients with rectal cancer after neoadjuvant CMT have no tumor cells beyond 1 cm from the residual tumor. Our data suggest that a 2-cm margin of resection when feasible from the macroscopic tumor margin after neoadjuvant CMT is adequate. PMID- 16269296 TI - Preoperative evaluation of pancreatic cystic lesions: cost-benefit analysis and proposed management algorithm. AB - BACKGROUND: This study was performed to develop a management algorithm that accurately predicts the necessity of operative intervention and results in significant cost savings for patients with cystic pancreatic tumors. METHODS: We reviewed 60 patients treated between 1992 and 2003. Accuracy rates of tests used to differentiate benign from premalignant and malignant cysts were calculated. A management algorithm was generated that incorporated clinical presentation, radiologic findings, and selective use of endoscopic procedures. This algorithm was tested for predictive accuracy, and savings between actual management and proposed management were compared. RESULTS: There was an average of 3 preoperative tests performed per patient. Endoscopic procedures included endoscopic retrograde cholangiopancreatography in 21 patients and endoscopic ultrasound in 25 patients. A cyst fluid carcinoembryonic antigen (CEA) level of 158 ng/mL or greater had an accuracy rate of 87.5% that was significantly higher than endoscopic retrograde cholangiopancreatography (72%), endoscopic ultrasound morphology (45%), or endoscopic ultrasound cytology (66.7%). There was no combination of tests that provided greater accuracy than cyst fluid CEA level alone (P < .05). The management algorithm had a positive predictive value of 81%. The average actual cost of radiographic and endoscopic procedures was dollar 8,080 per patient. The proposed cost based on the algorithm was dollar 6,677 with a savings of dollar 1,403 per patient (P = .009). CONCLUSIONS: In patients with clinical symptoms or radiographic findings of mucinous or malignant tumors, further testing is excessive. Of endoscopic tests available, the cystic fluid CEA level most accurately predicts the presence of a mucinous neoplasm. A management algorithm based on presenting symptoms, radiographic findings, and cyst fluid CEA level provides a guideline for the evaluation of cystic lesions in the most cost efficient manner while ensuring proper care. PMID- 16269295 TI - Natural history of indeterminate pancreatic cysts. AB - BACKGROUND: The optimal treatment for incidental asymptomatic pancreatic cysts is not known. The purpose of this study was to determine whether nonmucinous cysts by cyst-aspiration analysis can be observed safely. METHODS: A prospective protocol was initiated in September of 1999 for all suspected cystic neoplasms. Asymptomatic patients with negative cyst aspirates (no extracellular mucin, and concentration of carcinoembryonic antigen in the cyst fluid <200 ng/mL) were followed-up clinically and radiographically. RESULTS: Through December 2004, 221 patients have been evaluated, and 80 (36%) initially were operated. There were 141 (64%) patients with indeterminate cysts, 98 have been followed-up for more than 12 months. Compared with resected patients, observed patients were older (62 vs 56 y, P < .006), and had smaller cysts (2.4 vs 4.0, P = .001). At a mean follow-up period of 24 months, 4 patients (4%) were resected. The indication, time to resection, and pathology were as follows: 2 patients for symptoms (abdominal pain and obstructive jaundice) at 24 and 72 months, respectively: mucinous and serous cystadenomas; 1 patient for an increase in size (6.6 to 7.8 cm) at 18 months: lymphoepithelial cyst; and 1 patient for abdominal pain and increase in size (2.0 to 3.7 cm) at 41 months: pseudocyst. The only patient resected for a mucinous neoplasm had a cyst fluid carcinoembryonic antigen level of 896 ng/mL. In the remaining observed patients, 20 (23%) showed a decrease in cyst size, and 16 (19%) showed an increase in size (mean diameter change, 21%). CONCLUSIONS: Initial follow-up evaluation indicates that asymptomatic patients without evidence of a mucinous neoplasm by cyst aspiration can be followed clinically and with interval imaging. PMID- 16269297 TI - Outpatient minimally invasive parathyroidectomy using local/regional anesthesia: a safe and effective operative approach for selected patients. AB - BACKGROUND: Minimally invasive parathyroidectomy (MIP) using local/regional anesthesia has become an accepted treatment for selected patients with primary hyperparathyroidism (HPT) and can be performed in the ambulatory setting. METHODS: From 1999 to 2004, 139 consecutive patients at our institution with HPT caused by a single localized parathyroid adenoma underwent MIP through a 2.5- to 3-cm incision. Anesthesia included preoperative local/regional blocks with moderate intravenous sedation. Patient follow-up data were reviewed retrospectively. RESULTS: All 139 MIP patients had biochemical HPT and a single adenoma localized by sestamibi scan alone (n = 119; 86%) or combined with other imaging (n = 20; 14%). The mean adenoma size was 1,184 +/- 1,091 mg. Total calcium and parathyroid hormone levels were 11.3 +/- 0.8 mg/dL and 451 pg/mL preoperatively, respectively, decreasing to 9.4 +/- 0.6 mg/dL and 34 pg/mL postoperatively, respectively. Of MIP cases, 117 (84%) were completed with local/regional anesthesia, and 22 (16%) used general anesthesia (4 local/regional conversions). The mean operative time when reported was 56 +/- 21 minutes (n = 28). Same-day discharges occurred for 120 (86%) patients, whereas 16 patients were observed overnight and 3 patients were observed for 48 hours. Operative cure was achieved in 137 (98.6%) patients (follow-up period, 15.2 +/- 12.4 mo) with 1 morbidity (0.7%). CONCLUSIONS: Outpatient MIP is safe and effective in selected patients. A low morbidity (0.7% in this series), rapid recovery, and high biochemical cure rate (98.6%) parallels 4-gland exploration under general anesthesia. PMID- 16269298 TI - A description of morbidly obese state employees requesting a bariatric operation. AB - BACKGROUND: The federal government, the medical insurance industry, and the academic medical community have disagreed over what treatments are appropriate and cost effective for morbid obesity. This debate is hindered by inadequate data regarding the true costs of diseases and who chooses an operation as a treatment option. The purpose of this study was to obtain these costs and to describe this population. METHODS: Louisiana's managed medical insurance program created primarily for its civil service employees contracted to offer a small random group of morbidly obese employees the option of a bariatric operation. This observational study examined the subpopulation who requested consideration for the operation. We present historic cost data from all medical expenses paid by the insurance company, a telephone survey of the volunteers in the study to determine their medical problems, and diagnostic evaluation data on those employees randomized to proceed for possible bariatric operation. RESULTS: A total of 911 of 189,398 adult members of the insurance plan wanted to be considered for this study. Only 397, however, completed the informed-consent process. Of the 248 employees who met the age requirement, body mass index criteria, and health criteria to be considered for a bariatric operation and were randomized, 20 withdrew before obtaining 40 committed operative candidates. The 773 morbidly obese female members had used a mean of dollar 11,145 in medical insurance expenses in the year 2003 versus a mean of dollar 8,096 for the other 106,908 adult women. Similar values for the men were dollar 16,720 for the 138 morbidly obese men versus dollar 5,943 for the other 82,490 men. CONCLUSIONS: The morbidly obese members of this medical insurance plan who requested a bariatric operation are costing their plan 1.4 to 2.8 times the yearly amount of the other adult members in medical expenses. The yearly mean amount the insurance plan spends on these members suggests that operative treatment would pay for itself in a relatively few number of years if it could significantly reduce these costs. Even in those who consider bariatric operation, many withdraw, further limiting the costs of operative therapy. PMID- 16269299 TI - Survivin, a potential biomarker in the development of Barrett's adenocarcinoma. AB - BACKGROUND: Survivin, a member of the inhibitor-of-apoptosis family, is reported to be overexpressed in esophageal cancer but no data are available about its status in the metaplastic/dysplastic sequence. The aim of this study was to measure survivin gene expression in normal squamous/columnar epithelium and in the various stages of development of Barrett's adenocarcinoma. METHODS: Endoscopic biopsy or operative specimen samples from 5 tissue types were analyzed: (1) squamous epithelium from 3 cm above the gastroesophageal junction in patients with a negative pH study and no histologic injury (n = 17, pH- control); (2) antral tissue from patients with no evidence of Barrett's, dysplasia, or cancer (n = 29, antral control); (3) specialized intestinal metaplasia from patients with Barrett's esophagus (n = 16; Barrett's group); (4) low- or high-grade dysplasia (n = 12, dysplasia group), and (5) adenocarcinoma (n = 45 cancer group). After laser-capture microdissection cellular RNA was extracted from each tissue and reverse transcribed to complementary DNA. Expression levels of survivin were measured by reverse-transcription polymerase chain reaction. RESULTS: Survivin gene expression was greater in columnar (antral) compared with squamous (pH-) control tissues (P = .03). Expression in quiescent Barrett's epithelium was similar to both control tissues. Expression levels in dysplastic epithelium were greater than in squamous control (P = .01) and Barrett's tissues (P = .04), but not higher than columnar control tissues, whereas expression in adenocarcinoma was greater than all tissues except dysplasia (P < .001). CONCLUSIONS: Survivin expression may be a biomarker in the development of Barrett's adenocarcinoma that is able to distinguish between quiescent Barrett's, dysplastic Barrett's, and Barrett's adenocarcinoma. PMID- 16269300 TI - Perioperative outcomes and complications of laparoscopic ventral hernia repair. AB - BACKGROUND: Laparoscopic techniques are being used increasingly in the repair of ventral hernias and offer the potential benefits of a shorter hospital stay, decreased wound complications, and possibly a lower recurrence rate. Despite good results from high-volume centers, significant complications may occur with this approach and the morbidity of incisional hernia repair may be underestimated. The purpose of this study was to review our experience with laparoscopic ventral hernia repair (LVHR) since its inception at our institution. METHODS: Medical records of all patients who underwent LVHR at a single institution from May 2000 through December 2003 were reviewed. Preoperative and postoperative variables including complications were analyzed. Follow-up evaluation was by office visit and phone survey with assessment of patient satisfaction scores. Data are expressed as mean +/- SD. RESULTS: A total of 121 LVHR were performed in 116 patients (52 men, 64 women; mean age, 57 +/- 13 y; mean body mass index, 35 +/- 8). Hernias were recurrent in 35 cases (28.9%), with a mean of 1.4 prior repairs (range, 1-7). The mean defect size was 109 +/- 126 cm2 and the average mesh size used was 256 +/- 192 cm2. Operating time was 147 +/- 45 minutes, and the hospital stay averaged 1.7 +/- 1 days. Twelve cases (9.9%) were converted to open operation, most commonly because of extensive adhesions. Extensive laparoscopic adhesiolysis was necessary in 29 cases (26.6%). Overall, perioperative complications occurred in 33 cases (27.3%), 13 of which (39.3%) were persistent seromas. Major complications were seen in 9 cases (7.4%). There were 4 enterotomies (3.3%): 3 occurred as a result of adhesiolysis and 1 resulted from a trocar injury; 2 were detected intraoperatively and were converted to open operation and 2 presented postoperatively. One of these patients developed sepsis and died. Follow-up evaluation was available for 83.6% of cases at a mean interval of 22 +/- 16 months after repair. The hernia recurrence rate was 9.3% (9 cases) and was detected at a median of 6 months postoperatively. The overall patient satisfaction score was high at 4.3 +/- 1.1 (scale, 1-5). CONCLUSIONS: Laparoscopic repair is effective for the vast majority of patients with primary or recurrent ventral hernias and results in hernia recurrence rates of less than 10%, with high patient satisfaction scores. Although seroma is the most common complication, major morbidity occurred in 7.4% of the patients in our series. Enterotomy is the most common serious complication and may result in sepsis and death. PMID- 16269301 TI - Half-a-dozen ribs: the breakpoint for mortality. AB - BACKGROUND: We hypothesized that the number of rib fractures independently impacted patient pulmonary morbidity and mortality. METHODS: The National Trauma Data Bank (NTDB, v. 3.0 American College of Surgeons, Chicago, IL) was queried for patients sustaining 1 or more rib fractures. Data abstracted included the number of rib fractures by International Classification of Diseases-9 code, Injury Severity Score, the occurrence of pneumonia, acute respiratory distress syndrome, pulmonary embolus, pneumothorax, aspiration pneumonia, empyema, and associated injuries by abbreviated injury score, the need for mechanical ventilation, number of ventilator days, intensive care unit (ICU) length of stay (LOS), hospital LOS, mortality, and use of epidural analgesia. Statistical analysis was performed using the Student t test and linear regression analysis. Statistical significance was defined as a P value of less than .05. RESULTS: The NTDB included 731,823 patients. Of these, 64,750 (9%) had a diagnosis of 1 or more fractured ribs. Thirteen percent (n = 8,473) of those with rib fractures developed 13,086 complications, of which 6,292 (48%) were related to a chest-wall injury. Mechanical ventilation was required in 60% of patients for an average of 13 days. Hospital LOS averaged 7 days and ICU LOS averaged 4 days. The overall mortality rate for patients with rib fractures was 10%. The mortality rate increased (P < .02) for each additional rib fracture. The same pattern was seen for the following morbidities: pneumonia (P < .01), acute respiratory distress syndrome (P < .01), pneumothorax (P < .01), aspiration pneumonia (P < .01), empyema (P < .04), ICU LOS (P < .01), and hospital LOS for up to 7 rib fractures (P < .01). An association between increasing hospital LOS and number of rib fractures was not shown (P = .19). Pulmonary embolism also was not related to the number of rib fractures (P = .06). Epidural analgesia was used in 2.2% (n = 1,295) of patients with rib fractures. A reduction in mortality with epidural analgesia was shown at 2, 4, and 6 through 8 rib fractures. The use of epidural analgesia had no impact on the frequency of pulmonary complications. When stratifying data by Injury Severity Score and the presence or absence of rib fractures the mortality rates were similar. CONCLUSIONS: Increasing the number of rib fractures correlated directly with increasing pulmonary morbidity and mortality. Patients sustaining fractures of 6 or more ribs are at significant risk for death from causes unrelated to the rib fractures. Epidural analgesia was associated with a reduction in mortality for all patients sustaining rib fractures, particularly those with more than 4 fractures, but this modality of treatment appears to be underused. PMID- 16269302 TI - Biliary dyskinesia: the most common indication for cholecystectomy in children. AB - BACKGROUND: The purpose of this study is to examine the current indications for cholecystectomy in children and to evaluate the results after such surgery. METHODS: Retrospective analysis of 107 consecutive cholecystectomies performed in children at the Children's Hospital of Illinois between October 1998 and September 2003. Hospital medical charts and outpatient clinic charts were reviewed. Patients' families were contacted by telephone to obtain longer-term follow-up. Results were analyzed with SPSS 12.0 for Windows (SPSS Inc, Chicago, Ill). RESULTS: Biliary dyskinesia (BD) was the indication for surgery for 62 (58%) of the 107 children who underwent cholecystectomy during the study period. Gallbladder calculus (GC) disease was the next most common indication with 29 (27%) children. The duration of symptoms was longer for BD. The most common presenting symptom in both groups was abdominal pain. Food intolerance was reported by 45% of patients with BD, significantly higher than patients with GC. Mean length of stay after cholecystectomy was 17 hours and 45 hours for BD and GC, respectively. Short-term follow-up showed relief or improvement of symptoms in 85% of children with BD and in 97% with GC. There were no deaths. Two (1.9%) children of the total of 107 developed complications; both had intra-abdominal abscesses. Most patients had complete or considerable long-term improvement in symptoms. CONCLUSIONS: Biliary dyskinesia was the most common indication for cholecystectomy in children in our study. More than half of the surgeries were performed on an outpatient basis. Morbidity was minimal and mortality was zero. We had satisfactory short- and long-term symptom resolution with long-term patient satisfaction reaching 95%. PMID- 16269303 TI - Surgical treatment of hidradenitis suppurativa: a 10-year experience. AB - BACKGROUND: Hidradenitis suppurativa (HS) is a disease of the apocrine sweat glands resulting in chronic wounds with abscesses, sinuses, and fibrosis. Because many patients referred for treatment have both recurrent and progressive disability, we attempted to determine which factors have the greatest impact on outcome so we could develop an operative treatment algorithm. METHODS: We identified 57 patients with HS who underwent operative treatment for chronic recurrent HS from January 1994 through December 2003. Charts were reviewed for demographic, treatment, and outcome data. RESULTS: The mean age at presentation was 34 years and the average duration of symptoms was 6.7 years. Two thirds of the patients had undergone 1 or more incision and drainage procedures and 90% had received long-term antibiotic therapy. Axillary involvement was present in 88% of women and was bilateral in half of all patients. Inguinoperineal involvement was present in 87% of men and was bilateral in 92% of all patients. An algorithm for operative treatment was developed based on the extent of involvement, chronicity, and comorbid conditions. Ninety-two operative procedures were performed, 50% involved the axilla, 36% involved the perineum, and 14% involved the inguinal region. Excision and primary closure was used for localized disease; wide excision with or without skin grafting was used for diffuse disease. CONCLUSIONS: HS is a chronic relapsing disease that frequently causes disabling pain, diminished range of motion, and social isolation. Definitive treatment involves operative excision of the involved apocrine tissue and should be individualized based on the stage and location of the disease. PMID- 16269304 TI - Diagnosis and management of bile leaks after blunt liver injury. AB - BACKGROUND: Nonoperative management (NOM) of blunt liver injuries (BLIs) is common, but little data exist on the management of complications related to NOM. Furthermore, bile leaks are emerging as frequent complications of NOM. The goal of this study was to determine which BLI patients are at greatest risk for bile leaks and how to manage this complication. METHODS: Patients treated with NOM, including angiographic embolization (AE) and observation, were compared with those undergoing operation (OR). Mortality, hospital length of stay, liver related complications, and severity of liver injury were compared among the groups. RESULTS: Two hundred eighty-one patients sustained a BLI from 1997 through 2004. The Injury Severity Score and age of the groups were similar. The mortality rate of the OR patients was higher (P = .02). All bile leaks occurred in patients with a liver Abbreviated Injury Score (AIS) of > or = 4. For survivors of more than 4 days, 71% of OR, 50% of AE, and 17% of observed patients developed bile leaks. For the OR and AE groups, HIDA scanning detected all leaks, leading to earlier drainage procedures. Detection of a bile leak in NOM patients before 5 days of hospitalization led to shorter HLOS, P = .02. No patient with a negative HIDA scan developed a subsequent bile leak. CONCLUSIONS: NOM of BLIs is associated with lower mortality, but significant complications. Bile leaks are more common in patients with higher liver AIS, and in this study presented in only those with liver AIS > or = 4. HIDA scanning successfully diagnosed all patients with bile leaks and led to less invasive drainage procedures over time and shorter HLOS. PMID- 16269305 TI - The role of the lung in postinjury multiple organ failure. AB - BACKGROUND: Postinjury multiple organ failure (MOF) is a result of a dysfunctional inflammatory response to severe injury and shock. Acute lung injury is thought to promote further organ dysfunction by the systemic release of inflammatory mediators from injured lung tissue. Although clinical evidence supports this model, a clear understanding of the relationship between lung dysfunction and multiple organ failure has yet to be defined. We hypothesized that respiratory dysfunction is an early obligate event in the progression of postinjury MOF. METHODS: Data were collected prospectively on 1,344 trauma patients at risk for postinjury MOF. Inclusion criteria were age greater than 16 years, trauma intensive care unit admission, Injury Severity Score greater than 15, and survival longer than 48 hours. Isolated head injuries and head injuries with an extracranial abbreviated injury score of less than 2 were excluded. Daily physiologic and laboratory data were collected through surgical intensive care unit day 28 and clinical events were recorded thereafter until death or hospital discharge. Organ failure was characterized using the Denver MOF scale. RESULTS: Organ dysfunction was observed in 1,011 (75%) of 1,344 patients. Lung dysfunction was observed in 951 (94%) patients with 1 or more organ dysfunctions and 598 (99%) of 605 patients with 2 or more organ dysfunctions. Lung dysfunction preceded heart, liver, and kidney dysfunction by an average of 0.6 +/- 0.2 days, 4.8 +/- 0.2 days, and 5.5 +/- 0.5 days, respectively. The severity of lung dysfunction correlated with the severity of heart, liver, and kidney dysfunction, and the number of other dysfunctional organ systems. CONCLUSIONS: Postinjury respiratory dysfunction is an obligate event that precedes heart, liver, and kidney failure. The severity of other organ dysfunction is related directly to the severity of respiratory dysfunction. These data implicate lung dysfunction as central to the promotion of pathogenic inflammation and the development of postinjury MOF. PMID- 16269306 TI - Prospective analysis of the incidence of deep venous thrombosis in bariatric surgery patients. AB - BACKGROUND: Venous thromboembolism (VTE), manifest as deep venous thrombosis (DVT) or pulmonary embolus, remains an important complication in bariatric operation patients. Our purpose was to determine the incidence of VTE in a consecutive series of patients undergoing Roux-en-Y gastric bypass (RYGB) to guide appropriate therapy. METHODS: We prospectively examined a consecutive series of RYGB patients with bilateral lower-extremity venous duplex scan (DS) preoperatively, on postoperative day (POD)#2, and approximately POD#14. Preoperative clinical information including history of VTE, intraoperative data, postoperative course, and complications were recorded. Heparin 5,000 U subcutaneously was administered before the operation and every 12 hours throughout hospitalization along with sequential compression devices. Ambulation was instituted on POD#1. Temporary caval filters were placed in patients with a history of VTE. RESULTS: A total of 106 patients were examined. Body mass index was 51 +/- 8 kg/m2 (range, 40-73 kg/m2). Laparoscopic RYGB was performed in 75%. Hospital length of stay was 2.5 +/- 0.6 days. One hundred patients had no history of VTE; none had a positive DS preoperatively or on POD#2. One patient had a positive POD#14 DS and a second patient had a superficial thrombophlebitis, but a negative DS for DVT (both patients were symptomatic). Six patients had a prior history of VTE; all underwent preoperative placement of a temporary caval filter. Of these 6 patients, 1 developed a new postoperative DVT and another patient had thrombus on the caval filter with a negative lower-extremity DS. CONCLUSIONS: Occult DVT was not observed preoperatively in RYGB patients, suggesting that routine preoperative DS is not necessary in the absence of VTE history. Prophylaxis of heparin and sequential compression devices appears satisfactory in preventing DVT with only a 1% incidence in patients with no prior history of VTE. Two of the 6 patients with prior history of VTE showed evidence of thrombus postoperatively. Although a small number of patients, this finding suggests that a caval filter should be placed preoperatively in RYGB patients with a history of VTE. PMID- 16269307 TI - Anesthesia-specific protection from endotoxic shock is not mediated through the vagus nerve. AB - BACKGROUND: We have shown recently that volatile anesthetics significantly decrease inflammatory cytokine production and dramatically increase survival among rodents challenged with lipopolysaccharide (LPS). Because acetylcholine's interaction with nicotine receptors on tissue macrophages during vagus nerve stimulation has been implicated in the modulation of LPS-stimulated tumor necrosis factor alpha (TNF-alpha) production, we hypothesized that the mechanism of anesthetic immunoprotection is mediated through the vagus nerve. METHODS: Male Sprague-Dawley rats underwent bilateral cervical vagotomy (n = 20) or sham operation (n = 6). Twenty-four hours postoperatively, vagotomized rats were randomized into 3 groups: LPS injection (V+LPS, n = 6), LPS injection followed by 60 minutes of isoflurane anesthesia (V+LPS+ISO, n = 7), or saline injection (V+S, n = 7). Sham animals were also given LPS (Sham+LPS). A sublethal dose of LPS (8 mg/kg) was used. Blood samples were collected via cardiac puncture 90 minutes after LPS or saline injection, and plasma was isolated for the measurement of cytokines by enzyme-linked immunosorbent assay. Statistical differences between groups were detected by 1-way analysis of variance. RESULTS: Serum TNF-alpha was reduced significantly and interleukin (IL)-6 was abrogated completely among V+LPS+ISO rats, compared with both V+LPS and Sham+LPS animals (P < or = .05 for all). In contrast, levels of the anti-inflammatory cytokine IL-10 were similar among all LPS groups. CONCLUSIONS: Isoflurane anesthesia administered simultaneously with the injection of LPS decreases serum production of TNF-alpha and IL-6 despite bilateral transection of the vagus nerve. Isoflurane-mediated attenuation of proinflammatory cytokine production occurs via a mechanism other than modulation of vagal output. PMID- 16269308 TI - Preliminary results with early corticosteroid withdrawal in African American renal allograft recipients. AB - BACKGROUND: There is a paucity of data regarding the use of steroid-avoidance immunosuppression (SAI) in African American (AA) renal allograft recipients, traditionally considered a high-risk subgroup of patients with higher reported rates of acute rejection and graft loss. METHODS: We compared the outcomes of 27 AA renal allograft recipients receiving SAI (SA group; mean follow-up period, 12 +/- 3 mo) with those of 20 patients receiving a steroid taper (ST group; 24 +/- 11 mo). In both groups, thymoglobulin was used for induction, and mycophenolate mofetil and tacrolimus were used for maintenance. Four doses of methylprednisolone were given on days 0 to 3. In the SA group no further steroids were given, whereas in the ST group a prednisone taper was continued thereafter. RESULTS: ST patients were more likely to have current panel reactive antibody titers greater than 10%, undergo retransplantation, and receive more doses of thymoglobulin. There were no significant differences between the SA and ST groups with regard to patient survival (96% vs 95%), graft survival (96% vs 90%), acute rejection (11% vs 14%), cytomegalovirus infection (7% vs 10%), posttransplant diabetes mellitus (11% vs 24%), or mean serum creatinine concentration at 6 months (1.6 vs 1.5 mg/dL), respectively, with a trend toward less percent weight gain in SA patients at 6 months (5% vs 11%, P = .06). CONCLUSIONS: SAI can produce excellent short-term results in AA kidney transplant patients when compared with a conventional ST protocol. Our results will need to be verified in larger numbers of patients with longer follow-up evaluation. PMID- 16269309 TI - Vascular endothelial growth factor stimulates a novel calcium-signaling pathway in vascular smooth muscle cells. AB - BACKGROUND: Vascular endothelial growth factor (VEGF) is a potent vascular mitogen that selectively stimulates vascular smooth muscle cell (VSMC) migration through an unknown mechanism while having no effect on VSMC proliferation. It is known that VSMC migration and proliferation are dependent on the second messenger Ca2+ and, in particular, mitogen-stimulated Ca2+ influx. We hypothesized that the selective effect of VEGF on VSMC migration versus proliferation was a result of differential VEGF-stimulated Ca2+ signaling pathways. METHODS: Primary cultured human aortic smooth muscle cells (VSMCs) were grown to subconfluency and assigned to the following experimental groups: no stimulation, stimulation with platelet derived growth factor-BB (PDGF-BB) (20 ng/mL) as positive control, and stimulation with VEGF165 (40 ng/mL). Total increase in [Ca2+]cyt and intracellular calcium release was quantified with the use of a fura-2 fluorescence assay. Assays for the following receptors VEGFR-1 (Flt-1), VEGFR-2 (KDR/Flk-1) and PDGFR-beta were performed by immunoprecipitation, while PLCgamma1, Akt 1/2, and phospholamban B phosphorylation were assessed with Western immunoblotting. RESULTS: VSMCs stimulated with VEGF165 exhibited no intracellular Ca2+ release, compared with a 75 +/- 30 nmol/L intracellular calcium release after PDGF-BB stimulation, (P < .02) VEGF165-stimulated VSMCs in Ca2+-containing media exhibited 192 +/- 26 nmol/L increase in [Ca2+]cyt, compared with 354 +/- 54 nmol/L increase after PDGF-BB stimulation (P < .02). VEGF165 did not phosphorylate PLCgamma1 after 1, 5, or 10 minutes of treatment. VEGF165 treatment did not result in PI3-K/Akt activation at 1-, 5-, or 10-minute time points. Calmodulin-dependent kinase II (CaMKII) was activated by both VEGF165 and PDGF-BB after 1 and 5 minutes of stimulation. The presence of the receptors VEGFR 1, VEGFR-2, and PDGFR-beta was confirmed in all experimental groups. CONCLUSIONS: VEGF induces extracellular calcium influx but no intracellular calcium release in VSMCs. This lack of intracellular Ca2+ release stems from the inability of VEGF165 to activate the PLCgamma1 cascade and IP3 receptor-mediated Ca2+ release. The lack of PI3-K/Akt activation at these time points indicates a novel extracellular Ca2+ influx pathway sufficient to activate CaMKII. A paradigm relating extracellular Ca2+ influx to CaMKII activation and migration is suggested and may account for the selective effects of VEGF on VSMC migration. PMID- 16269310 TI - Expression of glial cell line-derived neurotrophic factor family members and their receptors in pancreatic cancers. AB - BACKGROUND: The glial cell line-derived neurotrophic factor (GDNF) is a member of neurotrophic polypeptide family, which promotes survival and rescue of various neural cells in the central and peripheral nerve systems. We previously reported that GDNF promotes tumor cell invasion in pancreatic cancer cell lines. The purpose of this study was to investigate GDNF family expression and the status of related receptors in actual cancer tissues, and assess correlations with clinicopathologic behavior. METHODS: Immunohistochemical assessment of GDNF, neurturin, persephin, artemin, GDNF family receptor alpha-1 and alpha-2, and RET was performed for 51 cases of surgically resected pancreatic cancer. RESULTS: In all intrapancreatic nerves, GDNF and artermin were expressed strongly. In pancreatic cancer tissues. The expression of RET was stronger than that seen in normal ductal cells and was significantly related to the survival rate after resection (P = .026) and lymphatic invasion (P = .014). Intrapancreatic neural invasion was significantly related to the expression of GDNF (P = .047). CONCLUSIONS: We conclude that the expression of RET in pancreatic cancer tissues may be a useful prognostic marker and GDNF may play an important role in neural invasion. PMID- 16269311 TI - Perfusion preservation maintains myocardial ATP levels and reduces apoptosis in an ex vivo rat heart transplantation model. AB - BACKGROUND: Machine perfusion preservation improves reperfusion function of many solid organs, compared with conventional storage, but has received limited clinical attention in preserving hearts for transplantation. We evaluated representative extracellular (Celsior) and intracellular (University of Wisconsion) storage solutions using static and perfusion protective strategies over a clinically relevant preservation period. METHODS: Rat hearts were preserved for 200 minutes by either static storage or perfusion preservation in Celsior or University of Wisconsin solutions. Three conditions were studied: conventional static storage; static storage using either solution with 5.5 mmol/L glucose added; and perfusion preservation using either solution with 5.5 mmol/L glucose added. Glucose was provided as U-13C-labeled glucose, and glycolysis and oxidative metabolism during preservation were quantified from incorporation of (13)C into glycolytic and tricarboxylic acid cycle intermediates. Adenosine triphosphate levels after preservation, and apoptosis and cardiac function after reperfusion were measured. RESULTS: Both perfusion preservation groups had higher myocardial oxygen consumption during storage and better early graft function, compared with static preservation groups (P < .05). Adenosine triphosphate levels were higher after storage in the perfusion groups (P < .01). Apoptosis was reduced in the perfusion groups (P < .01). Comparing perfusion groups, hearts preserved with Celsior had higher myocardial oxygen consumption and glucose utilization during perfusion storage and exhibited decreased reperfusion coronary vascular resistance and myocardial water content, compared with the UW perfusion group (P < .05). CONCLUSIONS: Perfusion preservation results in greater metabolism during storage and superior cardiac function with improved myocyte survival, compared with static storage. Extracellular preservation solutions appear more effective for perfusion preservation, possibly by augmenting cellular metabolism. PMID- 16269312 TI - Mitochondrial-dependent apoptosis in experimental rodent abdominal aortic aneurysms. AB - OBJECTIVES: While extrinsic mechanisms of apoptosis in abdominal aortic aneurysms (AAAs) are recognized, this project hypothesizes that an intrinsic, mitochondrial dependent, mechanism of apoptosis also contributes to experimental AAA formation. METHODS: Rat aortas were perfused with either saline or elastase (N = 5 per group) and harvested 7 days postperfusion. The aortas were placed in gluteraldehyde for subsequent transmission electron microscopy, Bouin's solution for TUNEL, or paraformaldehyde for immunohistochemical staining for caspase-9, caspase-3, and Bid. RESULTS: Abdominal aortic diameters increased 168 +/- 25% (mean +/- SEM) after elastase perfusion. compared with 30 +/- 5% after saline perfusion (P < .001). Apoptosis of aortic smooth muscle cells, macrophages, and neutrophils was evidenced by transmission electron microscopy and TUNEL in the elastase-perfused aneurysmal aortas. Quantitative analysis of the apoptotic cells revealed a significant (P < .01) increase in the number of total apoptotic cells in the elastase-perfused aortas (12 +/- 3 cells per high-power field), compared with that of saline-infused controls (1.3 +/- 0.2). Caspase-9, the key initiator in the mitochondrial-dependent apoptotic pathway, stained positively in only elastase-perfused aortas. Bid staining was not detected in either the elastase perfused aortas or the saline controls. CONCLUSIONS: Apoptosis is evident in multiple cell lines in elastase-perfused aneurysmal aortas, but rarely observed in control aortas. Caspase-9, the key initiator of intrinsic apoptosis, was documented only in elastase-perfused aortas. These results suggest that mitochondrial-dependent apoptosis is associated with abdominal aortic aneurysm formation. PMID- 16269314 TI - The importance of 'awareness' for understanding fetal pain. AB - Our understanding of when the fetus can experience pain has been largely shaped by neuroanatomy. However, completion of the cortical nociceptive connections just after mid-gestation is only one part of the story. In addition to critically reviewing evidence for whether the fetus is ever awake or aware, and thus able to truly experience pain, we examine the role of endogenous neuro-inhibitors, such as adenosine and pregnanolone, produced within the feto-placental unit that contribute to fetal sleep states, and thus mediate suppression of fetal awareness. The uncritical view that the nature of presumed fetal pain perception can be assessed by reference to the prematurely born infant is challenged. Rigorously controlled studies of invasive procedures and analgesia in the fetus are required to clarify the impact of fetal nociception on postnatal pain sensitivity and neural development, and the potential benefits or harm of using analgesia in this unique setting. PMID- 16269313 TI - Circadian regulation of sleep in mammals: role of the suprachiasmatic nucleus. AB - Despite significant progress in elucidating the molecular basis for circadian oscillations, the neural mechanisms by which the circadian clock organizes daily rhythms of behavioral state in mammals remain poorly understood. The objective of this review is to critically evaluate a conceptual model that views sleep expression as the outcome of opponent processes-a circadian clock-dependent alerting process that opposes sleep during the daily wake period, and a homeostatic process by which sleep drive builds during waking and is dissipated during sleep after circadian alerting declines. This model is based primarily on the evidence that in a diurnal primate, the squirrel monkey (Saimiri sciureus), ablation of the master circadian clock (the suprachiasmatic nucleus; SCN) induces a significant expansion of total daily sleep duration and a reduction in sleep latency in the dark. According to this model, the circadian clock actively promotes wake but only passively gates sleep; thus, loss of circadian clock alerting by SCN ablation impairs the ability to sustain wakefulness and causes sleep to expand. For comparison, two additional conceptual models are described, one in which the circadian clock actively promotes sleep but not wake, and a third in which the circadian clock actively promotes both sleep and wake, at different circadian phases. Sleep in intact and SCN-damaged rodents and humans is first reviewed, to determine how well the data fit these conceptual models. Neuroanatomical and neurophysiological studies are then reviewed, to examine the evidence for direct and indirect interactions between the SCN circadian clock and sleep-wake circuits. Finally, sleep in SCN-ablated squirrel monkeys is re examined, to consider its compatibility with alternative models of circadian regulation of sleep. In aggregate, the behavioral and neurobiological evidence suggests that in rodents and humans, the circadian clock actively promotes both wake and sleep, at different phases of the circadian cycle. The hypersomnia of SCN-ablated squirrel monkeys is unique in magnitude, but is not incompatible with a role for the SCN pacemaker in actively promoting sleep. PMID- 16269315 TI - Selective estrogen receptor modulators (SERMs) for the brain: current status and remaining challenges for developing NeuroSERMs. AB - Multiple issues regarding the efficacy of estrogen action in the brain remain unresolved. These include the timing, formulation and duration of the therapy intervention. Moreover, issues of thrombotic and neoplastic risks must be factored into the design of estrogen alternatives developed to prevent age associated neurodegenerative disorders, as well as other climacteric symptoms such as hot flush and sleep dysfunction. One strategy to address these issues is to develop molecules that selectively target and activate estrogen mechanisms of action in the brain while avoiding activation of estrogen receptors peripheral to the brain, particularly in reproductive organs. An overview of recent advances in our understanding of the molecular mechanisms of estrogen action is discussed in the context of designing an efficacious NeuroSERM that will activate cellular, biochemical and genomic events required for the promotion of memory function and neuronal survival. Pharmacological analyses of estrogen receptor subtypes and the case for a membrane-associated estrogen receptor splice variant in mediating these mechanisms are provided along with a summary of the activation profiles of existing clinically relevant estrogen alternatives or SERMs in neurons. Results of these endeavors have yielded insights into strategies for developing novel molecules with NeuroSERM potential in order to prevent brain related climacteric symptoms and neurodegenerative diseases. PMID- 16269316 TI - Interactions of alcohol and nitric-oxide synthase in the brain. AB - Nitric oxide (NO) is an important molecule associated with both physiological and pathological brain events. Three separate genes encode for nitric-oxide synthase (NOS), the rate-limiting enzyme in NO production, all of which are expressed within brain tissue. Effects of ethanol on NO production may be important to ethanol modification of brain function. Existing data indicate that alcohol exposure alters NOS expression and activity in the brain. Modulation of NOS is suggested to be involved in alcohol-induced behavioral modifications. Furthermore, alcohol-induced changes in NOS may alter immunocompetence, response to injury in the central nervous system, and may be involved in ethanol-mediated neurodegeneration and neurotoxicity. The extent and direction of change in NOS expression and activity depends on cell type and length of exposure. The mechanisms underlying these effects are only partially understood. Herein, the current understanding of the interactions of ethanol and NOS in the brain are discussed. PMID- 16269317 TI - The role of corticotropin-releasing factor-like peptides in cannabis, nicotine, and alcohol dependence. AB - The corticotropin-releasing factor (CRF)-like peptides, which include the mammalian peptides CRF, urocortin 1, urocortin 2, and urocortin 3, play an important role in orchestrating behavioral and physiological responses that may increase an organism's chance of survival when confronted with internal or external stressors. There is, however, evidence that a chronic overactivity of brain CRF systems under basal conditions may play a role in the etiology and maintenance of psychiatric disorders such as depression and anxiety disorders. In addition, there is evidence of a role for CRF-like peptides in acute and protracted drug abstinence syndromes and relapse to drug-taking behavior. This review focuses on the role of CRF-like peptides in the negative affective state associated with acute and protracted withdrawal from three widely abused drugs, cannabis, nicotine, and alcohol. In addition, we discuss the high comorbidity between stress-associated psychiatric disorders and drug dependence. A better understanding of the brain stress systems that may underlie psychiatric disorders, acute and protracted drug withdrawal, and relapse to drug-taking behavior may help in the development of new and improved pharmacotherapies for these widespread psychiatric disorders. PMID- 16269319 TI - The yin and yang of cardiac autonomic control: vago-sympathetic interactions revisited. AB - We review the pattern of activity in the parasympathetic and sympathetic nerves innervating the heart. Unlike the conventional textbook picture of reciprocal control of cardiac vagal and sympathetic nervous activity, as seen during a baroreceptor reflex, many other reflexes involve simultaneous co-activation of both autonomic limbs. Indeed, even at 'rest', the heart receives tonic drives from both sympathetic and parasympathetic cardiac nerves. Autonomic co-activation occurs during peripheral chemoreceptor, diving, oculocardiac, somatic nociceptor reflex responses as well as being evoked from structures within the brain. It is suggested that simultaneous co-activation may lead to a more efficient cardiac function giving greater cardiac output than activation of the sympathetic limb alone; this permits both a longer time for ventricular filling and a stronger contraction of the myocardium. This may be important when pumping blood into a constricted vascular tree such as is the case during the diving response. We discuss that in some instances, high drive to the heart from both autonomic limbs may also be arrhythmogenic. PMID- 16269318 TI - Lead neurotoxicity: from exposure to molecular effects. AB - The effects of lead (Pb(2+)) on human health have been recognized since antiquity. However, it was not until the 1970s that seminal epidemiological studies provided evidence on the effects of Pb(2+) intoxication on cognitive function in children. During the last two decades, advances in behavioral, cellular and molecular neuroscience have provided the necessary experimental tools to begin deciphering the many and complex effects of Pb(2+) on neuronal processes and cell types that are essential for synaptic plasticity and learning and memory in the mammalian brain. In this review, we concentrate our efforts on the effects of Pb(2+) on glutamatergic synapses and specifically on the accumulating evidence that the N-methyl-D-aspartate type of excitatory amino acid receptor (NMDAR) is a direct target for Pb(2+) effects in the brain. Our working hypothesis is that disruption of the ontogenetically defined pattern of NMDAR subunit expression and NMDAR-mediated calcium signaling in glutamatergic synapses is a principal mechanism for Pb(2+)-induced deficits in synaptic plasticity and in learning and memory documented in animal models of Pb(2+) neurotoxicity. We provide an introductory overview of the magnitude of the problem of Pb(2+) exposure to bring forth the reality that childhood Pb(2+) intoxication remains a major public health problem not only in the United States but worldwide. Finally, the latest research offers some hope that the devastating effects of childhood Pb(2+) intoxication in a child's ability to learn may be reversible if the appropriate stimulatory environment is provided. PMID- 16269320 TI - Protection from neuronal damage evoked by a motivational excitation is a driving force of intentional actions. AB - Motivation may be understood as an organism's subjective attitude to its current physiological state, which somehow modulates generation of actions until the organism attains an optimal state. How does this subjective attitude arise and how does it modulate generation of actions? Diverse lines of evidence suggest that elemental motivational states (hunger, thirst, fear, drug-dependence, etc.) arise as the result of metabolic disturbances and are related to transient injury, while rewards (food, water, avoidance, drugs, etc.) are associated with the recovery of specific neurons. Just as motivation and the very life of an organism depend on homeostasis, i.e., maintenance of optimum performance, so a neuron's behavior depends on neuronal (i.e., ion) homeostasis. During motivational excitation, the conventional properties of a neuron, such as maintenance of membrane potential and spike generation, are disturbed. Instrumental actions may originate as a consequence of the compensational recovery of neuronal excitability after the excitotoxic damage induced by a motivation. When the extent of neuronal actions is proportional to a metabolic disturbance, the neuron theoretically may choose a beneficial behavior even, if at each instant, it acts by chance. Homeostasis supposedly may be directed to anticipating compensation of the factors that lead to a disturbance of the homeostasis and, as a result, participates in the plasticity of motivational behavior. Following this line of thought, I suggest that voluntary actions arise from the interaction between endogenous compensational mechanisms and excitotoxic damage of specific neurons, and thus anticipate the exogenous compensation evoked by a reward. PMID- 16269321 TI - Glucose transport to the brain: a systems model. AB - Glucose transport to the brain involves sophisticated interactions of solutes, transporters, enzymes, and cell signaling processes, within an intricate spatial architecture. The dynamics of the transport are influenced by the adaptive nature of the blood-brain barrier (BBB), the semi-impermeable membranes of brain capillaries. As both the gate and the gatekeeper between blood-borne nutrients and brain tissue, the BBB helps govern brain homeostasis. Glucose in the blood must cross the BBB's luminal and abluminal membranes to reach neural tissue. A robust representation of the glucose transport mechanism can highlight a target for brain therapeutic intervention, help characterize mechanisms behind several disease phenotypes, or suggest a new delivery route for drugs. The challenge for researchers is understanding the relationships between influential physiological variables in vivo, and using that knowledge to predict how alterations or interventions affect glucose transport. This paper reviews factors influencing glucose transport and approaches to representing blood-to-brain glucose transport including in vitro, in vivo, and kinetic models. Applications for different models are highlighted, while their limitations in answering arising questions about the human in vivo BBB lead to a discussion of an alternate approach. A developing complex systems simulation is introduced, initiating a single platform to represent the dynamics of glucose transport across the adapting human blood brain barrier. PMID- 16269322 TI - Are mitochondria critical in the pathogenesis of Alzheimer's disease? AB - This review summarizes recent findings that suggest a causal connection between mitochondrial abnormalities and sporadic Alzheimer's disease (AD). Genetic causes of AD are known only for a small proportion of familial AD patients, but for a majority of sporadic AD patients, genetic causal factors are still unknown. Currently, there are no early detectable biomarkers for sporadic AD, and there is a lack of understanding of the pathophysiology of the disease. Findings from recent genetic studies of AD pathogenesis suggest that mitochondrial defects may play an important role in sporadic AD progression, and that mitochondrial abnormalities and oxidative damage may play a significant role in the progression of familial AD. Findings from biochemical studies, in vitro studies, gene expression studies, and animal model studies of AD are reviewed, and the possible contribution of mitochondrial mutations to late-onset sporadic AD is discussed. PMID- 16269323 TI - Ceruloplasmin in neurodegenerative diseases. AB - For decades, abnormalities in ceruloplasmin (Cp) synthesis have been associated with neurodegenerative disease. From the early observation that low circulating serum ceruloplasmin levels served as a marker for Wilson's disease to the recent characterization of a neurodegenerative disorder associated with a complete lack of serum ceruloplasmin, the link between Cp and neuropathology has strengthened. The mechanisms associated with these different central nervous system abnormalities are very distinct. In Wilson's disease, a defect in the P-type ATPase results in abnormal hepatic copper accumulation that eventually leaks into the circulation and is abnormally deposited in the brain. In this case, copper deposition results in the neurodegenerative phenotype observed. Patients with autosomal recessive condition, aceruloplasminemia, lack the ferroxidase activity inherent to the multi-copper oxidase ceruloplasmin and develop abnormal iron accumulation within the central nervous system. In the following review ceruloplasmin gene expression, structure and function will be presented and the role of ceruloplasmin in iron metabolism will be discussed. The molecular events underlying the different forms of neurodegeneration observed will be presented. Understanding the role of ceruloplasmin within the central nervous system is fundamental to further our understanding of the pathology observed. Is the ferroxidase function more essential than the antioxidant role? Does Cp help maintain nitrosothiol stores or does it oxidize critical brain substrates? The answers to these questions hold the promise for the treatment of devastating neurodegenerative conditions such as Alzheimer's and Parkinson's diseases. It is essential to further elucidate the mechanism of the neuronal injury associated with these disorders. PMID- 16269324 TI - Snaring the function of alpha-synuclein. AB - It is well established that the abundant neuronal protein alpha-synuclein has a causal role in Parkinson's disease, yet the normal functions of this protein remain unclear. In this issue of Cell, Chandra et al. (2005) reveal that alpha synuclein acts as a molecular chaperone, assisting in the folding and refolding of synaptic proteins called SNAREs. These proteins are crucial for release of neurotransmitters at the neuronal synapse, vesicle recycling, and synaptic integrity. PMID- 16269325 TI - The proteasome: not just degrading anymore. AB - The proteasome is a large multiprotein complex that has a critical role in the degradation of ubiquitylated proteins. A fascinating paper in this issue of Cell (Lee et al., 2005) now reveals that the proteasome recruits the SAGA histone acetyltransferase complex to a target promoter during gene activation. This finding adds to the growing body of evidence indicating that the proteasome has nonproteolytic functions. PMID- 16269326 TI - Hox genes: the instructors working at motor pools. AB - Motor neurons are assigned unique subidentities preceding their axon navigation. This ensures proper innervation of muscle targets and is accompanied by a stereotypical clustering of motor neuron cell bodies into "motor pools" within the spinal cord. However, the mechanisms that drive motor neuron diversification have been poorly understood. A new study by Dasen et al. (2005) in this issue of Cell shows that a network of Hox genes is responsible for instructing motor pool development. PMID- 16269327 TI - An E-MAP of the ER. AB - Genome-wide phenotype mapping has the potential to provide insights into gene function on an unprecedented scale. In this issue of Cell, Schuldiner et al.(2005) show that saturation epistasis analysis of genes implicated in the early secretory pathway in yeast provides a new strategy for uncovering functional relationships using high-throughput genomic data. PMID- 16269329 TI - Cohesin and the maternal age effect. AB - During meiosis in human oocytes, chromosome nondisjunction increases with maternal age, leading to disorders such as Down's syndrome. In a recent study in Nature Genetics, Hodges et al. (2005) show that mice with a mutation in the meiosis-specific cohesin protein SMC1beta exhibit age-dependent defects in meiosis. These defects are similar to those observed in oocytes of older human mothers. Their results implicate an age-dependent loss of function in SMC1beta (or related proteins) in the maternal age effect of humans. PMID- 16269328 TI - The genesis of a pandemic influenza virus. AB - Pandemic influenza viruses pose a significant threat to public health worldwide. In a recent Nature paper, Taubenberger et al. (2005) now report remarkable similarities between the polymerase genes of the influenza virus that caused the 1918 Spanish influenza pandemic and those of avian influenza viruses. Meanwhile, Tumpey et al. (2005) reporting in Science show that the reconstructed 1918 Spanish influenza virus kills mice faster than any other influenza virus so far tested. PMID- 16269330 TI - Membrane hemifusion: crossing a chasm in two leaps. AB - During membrane fusion, the outer leaflets of the two membranes merge first, whereas the distal membrane leaflets remain separate until the opening of a fusion pore. This intermediate stage, called hemifusion, is a critical event shared by exocytosis, protein trafficking, and viral entry. PMID- 16269331 TI - Alpha-synuclein cooperates with CSPalpha in preventing neurodegeneration. AB - Alpha-synuclein and cysteine-string protein-alpha (CSPalpha) are abundant synaptic vesicle proteins independently linked to neurodegeneration. Dominantly inherited mutations in alpha-synuclein cause Parkinson's disease, but the physiological role of alpha-synuclein remains unknown. Deletion of CSPalpha produces rapidly progressive neurodegeneration in mice, presumably because the cochaperone function of CSPalpha is essential for neuronal survival. Here, we report the surprising finding that transgenic expression of alpha-synuclein abolishes the lethality and neurodegeneration caused by deletion of CSPalpha. Conversely, ablation of endogenous synucleins exacerbates these phenotypes. Deletion of CSPalpha inhibits SNARE complex assembly; transgenic alpha-synuclein ameliorates this inhibition. In preventing neurodegeneration in CSPalpha deficient mice, alpha-synuclein does not simply substitute for CSPalpha but acts by a downstream mechanism that requires phospholipid binding by alpha-synuclein. These observations reveal a powerful in vivo activity of alpha-synuclein in protecting nerve terminals against injury and suggest that this activity operates in conjunction with CSPalpha and SNARE proteins on the presynaptic membrane interface. PMID- 16269332 TI - Chromosome morphogenesis: condensin-dependent cohesin removal during meiosis. AB - During meiosis, segregation of homologous chromosomes necessitates the coordination of sister chromatid cohesion, chromosome condensation, and recombination. Cohesion and condensation require the SMC complexes, cohesin and condensin, respectively. Here we use budding yeast Saccharomyces cerevisiae to show that condensin and Cdc5, a Polo-like kinase, facilitate the removal of cohesin from chromosomes prior to the onset of anaphase I when homologs segregate. This cohesin removal is critical for homolog segregation because it helps dissolve the recombination-dependent links between homologs that form during prophase I. Condensin enhances the association of Cdc5 with chromosomes and its phosphorylation of cohesin, which in turn likely stimulates cohesin removal. Condensin/Cdc5-dependent removal of cohesin underscores the potential importance of crosstalk between chromosome structural components in chromosome morphogenesis and provides a mechanism to couple chromosome morphogenesis with other meiotic events. PMID- 16269333 TI - The ubiquitin ligase HectH9 regulates transcriptional activation by Myc and is essential for tumor cell proliferation. AB - The Myc oncoprotein forms a binary activating complex with its partner protein, Max, and a ternary repressive complex that, in addition to Max, contains the zinc finger protein Miz1. Here we show that the E3 ubiquitin ligase HectH9 ubiquitinates Myc in vivo and in vitro, forming a lysine 63-linked polyubiquitin chain. Miz1 inhibits this ubiquitination. HectH9-mediated ubiquitination of Myc is required for transactivation of multiple target genes, recruitment of the coactivator p300, and induction of cell proliferation by Myc. HectH9 is overexpressed in multiple human tumors and is essential for proliferation of a subset of tumor cells. Our results suggest that site-specific ubiquitination regulates the switch between an activating and a repressive state of the Myc protein, and they suggest a strategy to interfere with Myc function in vivo. PMID- 16269334 TI - The proteasome regulatory particle alters the SAGA coactivator to enhance its interactions with transcriptional activators. AB - Promoter recruitment of the Saccharomyces cerevisiae SAGA histone acetyltransferase complex is required for RNA polymerase II-dependent transcription of several genes. SAGA is targeted to promoters through interactions with sequence-specific DNA binding transcriptional activators and facilitates preinitiation-complex assembly and transcription. Here, we show that the 19S proteasome regulatory particle (19S RP) alters SAGA to stimulate its interaction with transcriptional activators. The ATPase components of the 19S RP are required for stimulation of SAGA/activator interactions and enhance SAGA recruitment to promoters. Proteasomal ATPases genetically interact with SAGA, and their inhibition reduces global histone H3 acetylation levels and SAGA recruitment to target promoters in vivo. These results indicate that the 19S RP modulates SAGA complex using its ATPase components, thereby facilitating subsequent transcription events at promoters. PMID- 16269335 TI - Tumor suppressor HIC1 directly regulates SIRT1 to modulate p53-dependent DNA damage responses. AB - Hypermethylated in cancer 1 (HIC1) is an epigenetically regulated transcriptional repressor that functionally cooperates with p53 to suppress age-dependent development of cancer in mice. Here we show that the mechanism by which the loss of HIC1 function promotes tumorigenesis is via activating the stress-controlling protein SIRT1 and thereby attenuating p53 function. HIC1 forms a transcriptional repression complex with SIRT1 deacetylase, and this complex directly binds the SIRT1 promoter and represses its transcription. Inactivation of HIC1 results in upregulated SIRT1 expression in normal or cancer cells; this deacetylates and inactivates p53, allowing cells to bypass apoptosis and survive DNA damage. Inhibition of SIRT1 function in cells without HIC1 abolishes the resistance to apoptosis. Since aging increases promoter hypermethylation and epigenetic silencing of HIC1, we speculate that the resultant upregulation of SIRT1 may be a double-edged sword that both promotes survival of aging cells and increases cancer risk in mammals. PMID- 16269336 TI - Regulation of cell proliferation by a morphogen gradient. AB - One model to explain the relationship between patterning and growth during development posits that growth is regulated by the slope of morphogen gradients. The Decapentaplegic (DPP) morphogen controls growth in the Drosophila wing, but the slope of the DPP activity gradient has not been shown to influence growth. By employing a method for spatial, temporal, and quantitative control over gene expression, we show that the juxtaposition of cells perceiving different levels of DPP signaling is essential for medial-wing-cell proliferation and can be sufficient to promote the proliferation of cells throughout the wing. Either activation or inhibition of the DPP pathway in clones at levels distinct from those in surrounding cells stimulates nonautonomous cell proliferation. Conversely, uniform activation of the DPP pathway inhibits cell proliferation in medial wing cells. Our observations provide a direct demonstration that the slope of a morphogen gradient regulates growth during development. PMID- 16269337 TI - Calibrated measurement of gating-charge arginine displacement in the KvAP voltage dependent K+ channel. AB - Voltage-dependent ion channels open and conduct ions in response to changes in cell-membrane voltage. The voltage sensitivity of these channels arises from the motion of charged arginine residues located on the S4 helices of the channel's voltage sensors. In KvAP, a prokaryotic voltage-dependent K+ channel, the S4 helix forms part of a helical hairpin structure, the voltage-sensor paddle. We have measured the membrane depth of residues throughout the KvAP channel using avidin accessibility to different-length tethered biotin reagents. From these measurements, we have calibrated the tether lengths and derived the thickness of the membrane that forms a barrier to avidin penetration, allowing us to determine the magnitude of displacement of the voltage-sensor paddles during channel gating. Here we show that the voltage-sensor paddles are highly mobile compared to other regions of the channel and transfer the gating-charge arginines 15-20 A through the membrane to open the pore. PMID- 16269338 TI - A Hox regulatory network establishes motor neuron pool identity and target-muscle connectivity. AB - Spinal motor neurons acquire specialized "pool" identities that determine their ability to form selective connections with target muscles in the limb, but the molecular basis of this striking example of neuronal specificity has remained unclear. We show here that a Hox transcriptional regulatory network specifies motor neuron pool identity and connectivity. Two interdependent sets of Hox regulatory interactions operate within motor neurons, one assigning rostrocaudal motor pool position and a second directing motor pool diversity at a single segmental level. This Hox regulatory network directs the downstream transcriptional identity of motor neuron pools and defines the pattern of target muscle connectivity. PMID- 16269339 TI - Divergence of melanocortin pathways in the control of food intake and energy expenditure. AB - Activation of melanocortin-4-receptors (MC4Rs) reduces body fat stores by decreasing food intake and increasing energy expenditure. MC4Rs are expressed in multiple CNS sites, any number of which could mediate these effects. To identify the functionally relevant sites of MC4R expression, we generated a loxP-modified, null Mc4r allele (loxTB Mc4r) that can be reactivated by Cre-recombinase. Mice homozygous for the loxTB Mc4r allele do not express MC4Rs and are markedly obese. Restoration of MC4R expression in the paraventricular hypothalamus (PVH) and a subpopulation of amygdala neurons, using Sim1-Cre transgenic mice, prevented 60% of the obesity. Of note, increased food intake, typical of Mc4r null mice, was completely rescued while reduced energy expenditure was unaffected. These findings demonstrate that MC4Rs in the PVH and/or the amygdala control food intake but that MC4Rs elsewhere control energy expenditure. Disassociation of food intake and energy expenditure reveals unexpected divergence in melanocortin pathways controlling energy balance. PMID- 16269340 TI - Exploration of the function and organization of the yeast early secretory pathway through an epistatic miniarray profile. AB - We present a strategy for generating and analyzing comprehensive genetic interaction maps, termed E-MAPs (epistatic miniarray profiles), comprising quantitative measures of aggravating or alleviating interactions between gene pairs. Crucial to the interpretation of E-MAPs is their high-density nature made possible by focusing on logically connected gene subsets and including essential genes. Described here is the analysis of an E-MAP of genes acting in the yeast early secretory pathway. Hierarchical clustering, together with novel analytical strategies and experimental verification, revealed or clarified the role of many proteins involved in extensively studied processes such as sphingolipid metabolism and retention of HDEL proteins. At a broader level, analysis of the E MAP delineated pathway organization and components of physical complexes and illustrated the interconnection between the various secretory processes. Extension of this strategy to other logically connected gene subsets in yeast and higher eukaryotes should provide critical insights into the functional/organizational principles of biological systems. PMID- 16269341 TI - A slowed classical pathway rather than kiss-and-run mediates endocytosis at synapses lacking synaptojanin and endophilin. AB - The extent to which a "kiss-and-run" mode of endocytosis contributes to synaptic vesicle recycling remains controversial. The only genetic evidence for kiss-and run at the synapse comes from mutations in the genes encoding synaptojanin and endophilin, proteins that together function to uncoat vesicles in classical clathrin-mediated endocytosis. Here we have characterized the endocytosis that persists in null alleles of Drosophila synaptojanin and endophilin. In response to high-frequency stimulation, the synaptic-vesicle pool can be reversibly depleted in these mutants. Recovery from this depletion is slow and indicates the persistence of an impaired form of classical endocytosis. Steady-state exocytosis rates reveal that endocytosis saturates in mutant neuromuscular terminals at approximately 80 vesicles/s, 10%-20% of the wild-type rate. Analyses of quantal size, FM1-43 loading, and dynamin function further demonstrate that, even in the absence of synaptojanin or endophilin, vesicles undergo full fusion and re formation. Therefore, no genetic evidence remains to indicate that synaptic vesicles undergo kiss-and-run. PMID- 16269342 TI - Functional selection of a type IV pili-binding peptide that specifically inhibits Salmonella Typhi adhesion to/invasion of human monocytic cells. AB - Salmonella enterica serovar Typhi (S. Typhi) is an important pathogen which infects humans exclusively and causes typhoid or enteric fever. Recently it has been discovered that type IVB pili, encoded by the S. Typhi pil operon located in the major pathogenicity island, may be important in the pathogenesis of epidemic enteric fever. To further investigate the roles of type IVB pili of S. Typhi, a 12-mer peptide (RQERSSLSKPVV), binding to the structural protein PilS of the type IVB pili of S. Typhi, was isolated with a ribosome display system. This peptide was designated as peptide R. We found that peptide R inhibited adhesion to/invasion of human monocytic THP-1 cells by piliated S. Typhi bacteria, but had no effects on nonpiliated S. Typhi bacteria. A random 12-mer peptide, of size and solubility equal to peptide R, served as a control on the specificity of peptide R. The specific interaction and binding equilibrium between the 12-mer peptide R and PilS protein was determined by isothermal titration calorimetry (ITC) and a binding constant Ka determined to be between 0.4 x 10(5) and 2.2 x 10(5)L mol( 1). Our findings suggest that the type IV pili-binding peptide R holds potential as an antibacterial peptide effective against S. Typhi infections, both in terms of prevention and therapeutic treatment. The data further provide insights into the understanding of the pathogenic roles of the type IVB pili of S. Typhi. PMID- 16269343 TI - Diversity of wheat anti-microbial peptides. AB - From seeds of Triticum kiharae Dorof. et Migusch., 24 novel anti-microbial peptides were isolated and characterized by a combination of three-step HPLC (affinity, size-exclusion and reversed-phase) with matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry and Edman degradation. Based on sequence similarity and cysteine motifs, partially sequenced peptides were assigned to 7 families: defensins, thionins, lipid transfer proteins, hevein-like peptides, knottin-like peptides, glycine-rich peptides, and MBP-1 homologs. A novel subfamily of defensins consisting of 6 peptides and a new family of glycine-rich (8 peptides with different repeat motifs) were identified. Three 6-cysteine knottin-like peptides represented by N- and C-terminally truncated variants revealed no sequence homology to any known plant anti-microbial peptides. A new 8-cysteine hevein-like peptide and three 4 cysteine peptides homologous to MBP-1 from maize were isolated. This is the first communication on the occurrence of nearly all families of plant anti-microbial peptides in a single species. PMID- 16269344 TI - Lunatusin, a trypsin-stable antimicrobial peptide from lima beans (Phaseolus lunatus L.). AB - An anti-fungal peptide designated as lunatusin, with a molecular mass around 7kDa, was purified from the seeds of Chinese lima bean (Phaseolus lunatus L.). The peptide was isolated using a simple protocol consisting of affinity chromatography on Affi-gel blue gel and gel filtration on Superdex 75. Lunatusin exerted an anti-fungal activity toward fungal species such as Fusarium oxysporum, Mycosphaerella arachidicola and Botrytis cinerea, and an antibacterial action on, Bacillus megaterium, Bacillus subtilis, Proteus vulgaris and Mycobacterium phlei. It also inhibited proliferation in the breast cancer cell line MCF-7. Lunatusin reduced the activity of HIV-1 reverse transcriptase and it also inhibited translation in a cell-free rabbit reticulocyte lysate system. Its anti-fungal activity was retained after incubation with trypsin. Lunatusin elicited a mitogenic response from mouse splenocytes. PMID- 16269345 TI - In vitro activity of amphibian peptides alone and in combination with antimicrobial agents against multidrug-resistant pathogens isolated from surgical wound infection. AB - The in vitro activities of three amphibian peptides magainin II amide, citropin 1.1 and temporin A alone and in combination with eight clinically used antimicrobial agents (imipenem, ceftazidime, clarithromycin, vancomycin, amikacin, polymyxin E, ciprofloxacin and linezolid) were investigated against several multidrug-resistant Pseudomonas aeruginosa and Staphylococcus aureus strains isolated from surgical wound infections. Antimicrobial activities were measured by MIC, MBC and time-kill studies. P. aeruginosa strains were more susceptible to magainin II amide and less susceptible to temporin A. S. aureus isolates were highly susceptible to temporin A and citropin 1.1. The combination studies showed synergy between citropin 1.1 and clarithromycin. Magainin II amide and temporin A showed synergism with imipenem and ceftazidime. Finally, all peptides showed synergistic effects with polymyxin E. These results provide evidence for the potential use of these antimicrobial peptides in the topical or systemic treatment of surgical wound infections. PMID- 16269346 TI - Isolation and structural characterization of novel Rugosin A-like insulinotropic peptide from the skin secretions of Rana saharica frog. AB - Skin secretions of Rana saharica were evaluated for the isolation and characterization of novel insulinotropic peptides. Crude secretions obtained from young adult frogs by mild electrical stimulation of the dorsal skin surface were purified by reversed-phase HPLC yielding 80 fractions. In acute incubations with glucose-responsive BRIN-BD11cells, fractions 36-43, 46-54 and 57-63 showed the significant 2-8-fold increase in insulin-releasing activity (P<0.001) compared with 5.6mM of glucose alone. A pool of fractions 36-43 was subsequently rechromatographed to 28 homogenous peaks out of which 7 were capable of subsequent 1.5-3-fold increase in insulin release (P<0.001). Structural analysis of the non-toxic peptides with greatest insulin-releasing activity was performed by mass spectrometry and Edman degradation. Mass spectrometry analysis of two peaks indicated the molecular masses of 1892.6 and 2930.8Da. The sequence of the 1892.6-Da peptide was determined as KGAAKGLLEVASCKLSKSC, which has 68% homology with Rugosin A originally isolated from the skin secretion of Rana rugosa. A partial N-terminal sequence was determined for the 2930.8-Da peptide as AVITGACERDVQCGGGTCCAVSLI.... These data indicate that the skin secretions of Rana saharica frogs contain novel peptides with insulin-releasing activity. PMID- 16269347 TI - Identification of novel hexapeptide agonists at the Xenopus laevis melanophore melanocortin receptor. AB - We used a combinatorial chemical approach to identify novel agonists for the endogenous melanocortin receptor expressed in Xenopus laevis melanophores. A random one-bead one-compound hexapeptide library was screened to detect new molecules able to induce pigment dispersion in melanophores. Our approach led to the discovery of seven related novel peptides able to stimulate pigment dispersion with EC50 in the range of 0.1-10 microM. Their action was inhibited by the amphibian melanocortin receptor antagonist dWRL. These novel peptides share no significant sequence homology with known melanocortins. This study may aid in the understanding of the chemical interaction between the melanocortin receptors and their ligands. PMID- 16269348 TI - Characterization of a molt-inhibiting hormone (MIH) of the crayfish, Orconectes limosus, by cDNA cloning and mass spectrometric analysis. AB - The structure of the precursor of a molt-inhibiting hormone (MIH) of the American crayfish, Orconectes limosus was determined by cloning of a cDNA based on RNA from the neurosecretory perikarya of the X-organ in the eyestalk ganglia. The open reading frame includes the complete precursor sequence, consisting of a signal peptide of 29, and the MIH sequence of 77 amino acids. In addition, the mature peptide was isolated by HPLC from the neurohemal sinus gland and analyzed by ESI-MS and MALDI-TOF-MS peptide mapping. This showed that the mature peptide (Mass 8664.29 Da) consists of only 75 amino acids, having Ala75-NH2 as C terminus. Thus, C-terminal Arg77 of the precursor is removed during processing, and Gly76 serves as an amide donor. Sequence comparison confirms this peptide as a novel member of the large family, which includes crustacean hyperglycaemic hormone (CHH), MIH and gonad (vitellogenesis)-inhibiting hormone (GIH/VIH). The lack of a CPRP (CHH-precursor related peptide) in the hormone precursor, the size and specific sequence characteristics show that Orl MIH belongs to the MIH/GIH(VIH) subgroup of this larger family. Comparison with the MIH of Procambarus clarkii, the only other MIH that has thus far been identified in freshwater crayfish, shows extremely high sequence conservation. Both MIHs differ in only one amino acid residue ( approximately 99% identity), whereas the sequence identity to several other known MIHs is between 40 and 46%. PMID- 16269349 TI - Mass spectrometric characterization of crustacean hyperglycemic hormone precursor related peptides (CPRPs) from the sinus gland of the crab, Cancer productus. AB - Crustacean hyperglycemic hormone (CHH) precursor-related peptides (CPRPs) are produced during the proteolytic processing of CHH preprohormones. Currently, the physiological roles played by CPRPs are unknown. Due to their large size, direct mass spectrometric sequencing of intact CPRPs is difficult. Here, we describe a novel strategy for sequencing Cancer productus CPRPs directly from a tissue extract using nanoflow liquid chromatography coupled to quadrupole time-of-flight tandem mass spectrometry. Four novel CPRPs were characterized with the aid of MS/MS de novo sequencing of 27 truncated CPRP peptides. Extensive modifications (methionine oxidation and carboxy-terminal methylation) were identified in both the full-length and truncated peptides. To investigate the origin of the modifications and truncations, a full-length CPRP was synthesized and subjected to the same storage and extraction protocols used for the characterization of the native peptides. Here, some methionine oxidation was seen, however, no methylation or truncation was evident suggesting much of the chemical complexity seen in the native CPRPs is unlikely due to a sample preparation artifact. Collectively, our study represents the most complete characterization of CPRPs to date and provides a foundation for future investigation of CPRP function in C. productus. PMID- 16269350 TI - Rapid presymptomatic detection of PrPSc via conformationally responsive palindromic PrP peptides. AB - Structurally unique, synthetic prion peptides provide the basis of a simple assay to serve as both a detection and signal amplification system that distinguishes the normal prion protein, PrPC, from the misfolded prion protein, PrPSc, that is associated with the occurrence of transmissible spongiform encephalopathies (TSE). Proof-of-principle has been shown on brain samples from an experimental scrapie hamster model. The assay demonstrates very sensitive detection of PrPSc in animal brain tissue with potential application for early presymptomatic detection in animal screening. Furthermore, the sensitivity of the assay could enable blood tests for this TSE disease as well as other amyloid and/or misfolded protein diseases. PMID- 16269351 TI - Insulin-like growth factor (IGF) induced proliferation of human lung fibroblasts is enhanced by neurotensin. AB - Fibroblasts are key cells in tissue repair and important contributors to the inflammatory response. Insulin-like growth factors (IGFs) have been shown to participate in growth, in immune responses and in tissue repair where they stimulate cell growth. Neurotensin (NT) has been suggested to participate in inflammation and in tissue repair and is an autocrine or paracrine growth factor for several cancer cell types. Here we show that IGF-induced proliferation of fibroblasts is enhanced by NT in a concentration and type 1 NT-receptor dependent manner. This action of NT was blocked by inhibitors of phospholipase C and protein kinase C but not by inhibitors of phosphoinositide-3-kinase. An inhibitor of MEK 1/2 significantly reduced the proliferative effects of the IGFs but NT's ability to enhance IGF-induced proliferation was not effected. The ability of NT to enhance IGF-induced proliferation did not involve an autocrine factor. These results suggest that interactions between NT and the IGFs may contribute to the regulation of fibroblasts in for example, inflamed or injured tissues. PMID- 16269352 TI - Urotensin II: evidence for cardiac, hepatic and renal production. AB - Although urotensin II (UII) has been reported to circulate in human plasma and be raised in cardiovascular disorders, little, if any, information is available regarding the source of plasma UII. Accordingly, we have performed trans-organ arteriovenous sampling for measurement of UII concentration in anesthetized sheep. Plasma UII levels were measured in the low picomolar range in normal sheep and arterial plasma levels rose steadily with increasing time of anesthesia. Significant arteriovenous gradients were observed across the heart (36%), liver (40%) and kidney (44%). This is the first report to identify the heart, liver and kidney as sources of UII in the circulation. PMID- 16269353 TI - Doxapram increases corticotropin-releasing factor immunoreactivity and mRNA expression in the rat central nucleus of the amygdala. AB - Doxapram causes panic anxiety in humans. To determine whether doxapram alters corticotropin-releasing factor (CRF) expression in the central nucleus of the amygdala (CeA), paraventricular nucleus of hypothalamus (PVN), or bed nucleus of the stria terminalis (BNST), we used immunohistochemistry to measure CRF peptide in these brain areas after doxapram injection. Doxapram injection significantly increased CRF-like immunoreactivity (CRF-IR) within the CeA, but not in the BNST or PVN, and this increase was significant 2h after injection. In addition, doxapram significantly increased CRF mRNA expression within the CeA, and this was most prominent 30min after injection. These results suggest that doxapram selectively increases CRF expression within the CeA, and that this is mediated by increased CRF gene transcription. This increase in CRF-IR within the CeA might explain the doxapram-induced anxiety reaction. PMID- 16269354 TI - Evidence of involvement of leptin and IL-6 peptides in the action of interferon beta in secondary progressive multiple sclerosis. AB - Leptin is a peptide hormone which acts on cells of immune system by influencing the production of cytokines. Serum leptin levels and cytokine production by peripheral blood mononuclear cells (PBMC) were measured in 18 secondary progressive multiple sclerosis (SPMS) patients under IFN-beta-1b treatment. There were no overall effects on leptin, interleukin-6 (IL-6), IL-10 and IL-12 p40 after 2, 6 and 12 months of treatment. However, leptin and IL-6 decreased after 6 and 12 months of treatment in 12 patients who did not show progression of disability. Thus, our pilot data show that the beneficial effect of IFN-beta on some SPMS patients might be associated with the reduced levels of leptin and reduced IL-6 production by PBMC. PMID- 16269355 TI - Central administration of peptide and small molecule MC4 receptor antagonists induce hyperphagia in mice and attenuate cytokine-induced anorexia. AB - We investigated the effect of melanocortin 4 receptor (MC4) antagonists on food intake in mice. Food intake during the light phase was significantly increased by ICV administration of mixed MC3/MC4 antagonists (AgRP and SHU9119) or MC4 selective antagonist peptide [(Cyclo (1-5)[Suc-D-Nal-Arg-Trp-Lys]NH2] (MBP10) and the small molecule antagonists THP and NBI-30. Both mixed and selective antagonists significantly reversed anorexia induced by ICV administration of the MC4 agonist (c (1-6) HfRWK-NH2) and the cytokine IL-1beta. These findings provide pharmacological evidence that the MC4 receptor mediates the effects of melanocortin agonists and antagonists on food intake in mice, and support the idea that selective small molecule MC4 antagonists may be useful as therapeutics for cachexia. PMID- 16269356 TI - CCK and 5-HT act synergistically to suppress food intake through simultaneous activation of CCK-1 and 5-HT3 receptors. AB - Cholecystokinin (CCK) and serotonin (5-HT) systems have been shown to cooperate interdependently in control of food intake. To assess mechanisms by which CCK and 5-HT systems interact in control of food intake we examined: (1) participation of CCK-1 and 5-HT3 receptors in 5-HT-induced suppression of sucrose intake; (2) the interaction between CCK and 5-HT in suppression of food intake; (3) the role of CCK-1 and 5-HT3 receptors in mediating this interaction. Intraperitoneal administration of 5-HT (0.25, 0.5 and 1.0 mg/kg) significantly reduced intake compared to control in a dose responsive fashion (r2=0.989). Suppression of food intake by 5-HT was significantly attenuated by prior treatment with the 5-HT3 receptor antagonist ondansetron at each 5-HT dose tested (P<0.05), while blockade of CCK-1 receptors by lorglumide had no effect on 5-HT-induced suppression of intake. Administration of CCK-8 (0.5 microg/kg) or 5-HT (0.5 mg/kg) alone significantly reduced sucrose intake by 22.9 and 22.2% respectively, compared to control (P<0.0001). Co-administration of CCK and 5-HT resulted in a synergistic suppression of intake leading to an overall 48.4% reduction in sucrose intake compared to saline (P<0.0001). Concomitant CCK-1 and 5-HT3 receptor blockade by lorglumide and ondansetron respectively, resulted in a complete reversal of the combined CCK and 5-HT-induced suppression of intake. Independent administration of lorglumide or ondansetron did not alter intake compared to control. These studies provide evidence that 5-HT causes suppression in food intake by acting at 5-HT3, not CCK-1 receptors. Furthermore, CCK and 5-HT interact to produce an enhanced suppression of food intake, an effect mediated through concomitant activation of CCK-1 and 5-HT3 receptors. PMID- 16269358 TI - Beyond counting photons: trials and trends in vertebrate visual transduction. AB - For over 30 years, photoreceptors have been an outstanding model system for elucidating basic principles in sensory transduction and G protein signaling. Recently, photoreceptors have become an equally attractive model for studying many facets of neuronal cell biology. The primary goal of this review is to illustrate this rapidly growing trend. We will highlight the areas of active research in photoreceptor biology that reveal how different specialized compartments of the cell cooperate in fulfilling its overall function: converting photon absorption into changes in neurotransmitter release. The same trend brings us closer to understanding how defects in photoreceptor signaling can lead to cell death and retinal degeneration. PMID- 16269359 TI - Hair-cell mechanotransduction and cochlear amplification. AB - In the inner ear, sensory hair cells not only detect but also amplify the softest sounds, allowing us to hear over an extraordinarily wide intensity range. This amplification is frequency specific, giving rise to exquisite frequency discrimination. Hair cells detect sounds with their mechanotransduction apparatus, which is only now being dissected molecularly. Signal detection is not the only role of this molecular network; amplification of low-amplitude signals by hair bundles seems to be universal in hair cells. "Fast adaptation," the rapid closure of transduction channels following a mechanical stimulus, appears to be intimately involved in bundle-based amplification. PMID- 16269360 TI - Olfaction: diverse species, conserved principles. AB - Olfaction is a vitally important sense for all animals. There are striking similarities between species in the organization of the olfactory pathway, from the nature of the odorant receptor proteins, to perireceptor processes, to the organization of the olfactory CNS, through odor-guided behavior and memory. These common features span a phylogenetically broad array of animals, implying that there is an optimal solution to the problem of detecting and discriminating odors. PMID- 16269361 TI - Humans as an animal model for systems-level organization of olfaction. AB - The past 15 years have seen significant advances in the study of olfaction, with particular emphasis on elucidating the molecular building blocks of the sensory process. However, much of the systems-level organization of olfaction remains unexplored. Here, we provide an overview at this level, highlighting results obtained from studying humans, whom we think provide an underutilized, yet critical, animal model for olfaction. PMID- 16269362 TI - Taste recognition: food for thought. AB - The ability to identify food that is nutrient-rich and avoid toxic substances is essential for an animal's survival. Although olfaction and vision contribute to food detection, the gustatory system acts as a final checkpoint control for food acceptance or rejection behavior. Recent studies with model organisms such as mice and Drosophila have identified candidate taste receptors and examined the logic of taste coding in the periphery. Despite differences in terms of gustatory anatomy and taste-receptor families, these gustatory systems share a basic organization that is different from other sensory systems. This review will summarize our current understanding of taste recognition in mammals and Drosophila, highlighting similarities and raising several as yet unanswered questions. PMID- 16269363 TI - A comparison of experience-dependent plasticity in the visual and somatosensory systems. AB - In the visual and somatosensory systems, maturation of neuronal circuits continues for days to weeks after sensory stimulation occurs. Deprivation of sensory input at various stages of development can induce physiological, and often structural, changes that modify the circuitry of these sensory systems. Recent studies also reveal a surprising degree of plasticity in the mature visual and somatosensory pathways. Here, we compare and contrast the effects of sensory experience on the connectivity and function of these pathways and discuss what is known to date concerning the structural, physiological, and molecular mechanisms underlying their plasticity. PMID- 16269364 TI - Reliability and representational bandwidth in the auditory cortex. AB - It is unclear why there are so many more neurons in sensory cortex than in the sensory periphery. One possibility is that these "extra" neurons are used to overcome cortical noise and faithfully represent the acoustic stimulus. Another possibility is that even after overcoming cortical noise, there is "excess representational bandwidth" available and that this bandwidth is used to represent conjunctions of auditory and nonauditory information for computation. Here, we discuss recent data about neuronal reliability in auditory cortex showing that cortical noise may not be as high as was previously believed. Although at present, the data suggest that auditory cortex neurons can be more reliable than those in the visual cortex, we speculate that the principles governing cortical computation are universal and that visual and other cortical areas can also exploit strategies based on similarly high-fidelity activity. PMID- 16269365 TI - Why seeing is believing: merging auditory and visual worlds. AB - Vision may dominate our perception of space not because of any inherent physiological advantage of visual over other sensory connections in the brain, but because visual information tends to be more reliable than other sources of spatial information, and the central nervous system integrates information in a statistically optimal fashion. This review discusses recent experiments on audiovisual integration that support this hypothesis. We consider candidate neural codes that would enable optimal integration and the implications of optimal integration for perception and plasticity. PMID- 16269366 TI - Vagaries of visual perception in autism. AB - Three classes of perceptual phenomena have repeatedly been associated with autism spectrum disorder (ASD): superior processing of fine detail (local structure), either inferior processing of overall/global structure or an ability to ignore disruptive global/contextual information, and impaired motion perception. This review evaluates the quality of the evidence bearing on these three phenomena. We argue that while superior local processing has been robustly demonstrated, conclusions about global processing cannot be definitively drawn from the experiments to date, which have generally not precluded observers using more local cues. Perception of moving stimuli is impaired in ASD, but explanations in terms of magnocellular/dorsal deficits do not appear to be sufficient. We suggest that abnormalities in the superior temporal sulcus (STS) may provide a neural basis for the range of motion-processing deficits observed in ASD, including biological motion perception. Such an explanation may also provide a link between perceptual abnormalities and specific deficits in social cognition associated with autism. PMID- 16269367 TI - Neurocognitive mechanisms of synesthesia. AB - Synesthesia is a condition in which stimulation of one sensory modality causes unusual experiences in a second, unstimulated modality. Although long treated as a curiosity, recent research with a combination of phenomenological, behavioral, and neuroimaging methods has begun to identify the cognitive and neural basis of synesthesia. Here, we review this literature with an emphasis on grapheme-color synesthesia, in which viewing letters and numbers induces the perception of colors. We discuss both the substantial progress that has been made in the past fifteen years and some open questions. In particular, we focus on debates in the field relating to the neural basis of synesthesia, including the relationship between synesthesia and attention and the role of meaning in synesthetic colors. We propose that some, but probably not all, of these differences can be accounted for by differences in the synesthetes studied and discuss some methodological implications of these individual differences. PMID- 16269368 TI - Gadolinium-enhanced computed tomographic angiography: current status. AB - This article reviews the research to date, as well as our clinical experience from two institutions, on gadolinium-enhanced computed tomographic angiography (gCTA) for imaging the body. gCTA may be an appropriate examination for the small percentage of patients who would benefit from noninvasive vascular imaging, but who have contraindications to both iodinated contrast and magnetic resonance imaging. gCTA is more expensive than CTA with iodinated contrast, due to the dose of gadolinium administered, and gCTA has limitations compared with CTA with iodinated contrast, in that parenchymal organs are not optimally enhanced at doses of 0.5 mmol/kg or lower. However, in our experience, gCTA has been a very useful problem-solving examination in carefully selected patients. With the advent of 16-64 detector CT, in combination with bolus tracking, we believe that the overall dose of gadolinium needed for diagnostic CTA examinations, while relatively high, can be safely administered. PMID- 16269369 TI - Musculoskeletal imaging with computed tomography and magnetic resonance imaging: when is computed tomography the study of choice? AB - Indications for computed tomography (CT) of the musculoskeletal system, especially with the advent of 16-slice multidetector CT (MDCT), are numerous. In addition to the evaluation of the trauma patient where CT is essential when imaging complex skeletal injuries, MDCT is particularly useful in patients who have had prior surgery. In postoperative cases, metal artifact typically prohibits magnetic resonance imaging evaluation, but volume-rendering of a MDCT axial database virtually eliminates streak artifact associated with hardware. For the evaluation of masses, CT provides the ability to detect and characterize calcification, cortical disruption, and periosteal reaction. In this article, these and other indications for performance of CT of the musculoskeletal system will be discussed. PMID- 16269370 TI - The placenta revisited: radiologic-pathologic correlation. AB - The placenta is the central support organ for the developing fetus. Recognition of placental variants and insignificant findings is important so as not to suggest an abnormality when one is not present. However, the degree of abnormality, as well as the clinical implications of the findings, must be understood to help guide management of the pregnancy. This article reviews the normal sonographic appearance of the placenta and some anatomic variants, in addition to illustrating various common pathological conditions of the placenta and correlating with gross pathologic and histologic specimens. PMID- 16269371 TI - 2005 SSAT presidential address: Heroes, Mentors, and Teammates. PMID- 16269372 TI - Robotic-assisted Heller myotomy versus laparoscopic Heller myotomy for the treatment of esophageal achalasia: multicenter study. AB - Laparoscopic Heller myotomy (LHM) has become the standard treatment option for achalasia. The incidence of esophageal perforation reported is about 5%-10%. Robotically assisted Heller myotomy (RAHM) is emerging as a safe alternative to LHM. Data comparing the two approaches are scant. The aim of this study was to compare RAHM with LHM in terms of efficacy and safety for treatment of achalasia. A total of 121 patients underwent surgical treatment of achalasia at three institutions. A retrospective review of prospectively collected perioperative data was performed. Patients were divided into two groups: group A (RAHM), 59 patients, and group B (LHM), 62 patients. All the operations were completed using minimally invasive techniques. There were 63 women and 58 men, with a mean age of 45 +/- 19 years (14-82 years). Fifty-one percent of patients in group A and 95% of patients in group B reported weight loss. Duration of symptoms was equal for both groups. Dysphagia was the main complaint in both groups (P = NS). There was no difference in preoperative endoscopic treatment in both groups (44% versus 27%, P = NS). Operative time was significantly shorter for LHM in the first half of the experience (141 +/- 49 versus 122 +/- 44 minutes, P < .05). However, in the last 30 cases there was no difference in operative time between the groups (P = NS). Intraoperative complications (esophageal perforation) were more frequent in group B (16% versus 0%). The incidence of postoperative heartburn did not differ by group. There were no deaths. At 18 and 22 months, 92% and 90% of patients had relief of their dysphagia. This study suggests that RAHM is safer than LHM, because it decreases the incidence of esophageal perforation to 0%, even in patients who had previous treatment. At short-term follow-up, relief of dysphagia was equally achieved in both groups PMID- 16269373 TI - Esophagogastrectomy: the influence of stapled versus hand-sewn anastomosis on outcome. AB - Successful anastomosis is essential for favorable esophagogastrectomy outcomes. Before July 2002, almost all esophagogastric anastomoses at our institution were hand-sewn. We then began using linear stapled anastomotic techniques. This review compares patient outcomes with both techniques. From July 2001 to June 2004, 280 consecutive esophagogastrectomy patients (235 men and 45 women) were reviewed (median age, 65 years). The anastomosis was intrathoracic in 206 patients (74%) and cervical in 74 (26%). Anastomoses were hand-sewn in 205 patients (73%) and linear stapled in 75 (27%). Stapled anastomoses were intrathoracic in 33 patients (16%) and cervical in 42 (57%). Anastomotic leaks occurred in 30 patients (11%); 26 (12.7%) in the hand-sewn and 4 (5.3%) in the linear stapled group (P = .008). Leaks were asymptomatic in 17 patients (57%). Dilatation was required in 70 hand sewn anastomoses (34%) and in 11 stapled (14.6%) (P = .001). Hand-sewn anastomoses were more likely to leak and require dilatation; odds ratios and 95% confidence intervals were 5.35 (1.67-19.27) and 3.58 (1.66-8.34), respectively. A linear stapled anastomosis is safe and associated with both a significantly lower leak rate and the need for dilatation compared with hand-sewn anastomosis. This nonrandomized series suggests that linear stapled anastomosis is the preferred technique regardless of anastomotic location. PMID- 16269375 TI - Idiopathic pulmonary fibrosis: how often is it really idiopathic? AB - The cause of idiopathic pulmonary fibrosis (IPF) is unknown. The pathology suggests that IPF results from serial lung injury. It has been suggested that gastroesophageal reflux disease (GERD) may relate to the cause or the progression of the disease. The aims of this study were to determine the prevalence of GERD, the clinical presentation of GERD, and the manometric and reflux profiles in patients with end-stage IPF. Between July 2003 and October 2004, 18 patients with IPF on the lung transplant waiting list were referred for evaluation to the Swallowing Center of the University of California San Francisco. On the basis of the results of the pH monitoring test (5 and 20 cm above the lower esophageal sphincter), the patients were divided into two groups: group A, 12 patients (66%), GERD+; group B, 6 patients (34%), GERD-. The incidence of heartburn and regurgitation was similar between GERD+ and GERD- patients; reflux was clinically silent in one third of GERD+ patients. Reflux was associated with a hypotensive lower esophageal sphincter and abnormal esophageal peristalsis, and it was present in the upright and supine position. The reflux often extended into the proximal esophagus. These results show the following: (1) Two thirds of patients with IPF had GERD; (2) symptoms could not distinguish between those with and without GERD; (3) reflux occurred in the presence of a hypotensive lower esophageal sphincter and abnormal esophageal peristalsis; and (4) reflux occurred in the upright and supine positions, and often extended into the proximal esophagus. We conclude that patients with IPF should be screened for GERD, and if GERD is present, a fundoplication should be performed before or shortly after lung transplantation. PMID- 16269374 TI - Forty-eight-hour pH monitoring increases sensitivity in detecting abnormal esophageal acid exposure. AB - Ambulatory 24-hour esophageal pH measurement is the standard for detecting abnormal esophageal acid exposure (AEAE), but it has a false negative rate of 15% to 30%. Wireless 48-hour pH monitoring (Bravo; Medtronic, Shoreview, MN) may allow more accurate detection of AEAE versus 24-hour pH monitoring. Forty-eight hour wireless data were reviewed from 209 patients at three different tertiary care referral centers between 2003 and 2005. Manometric or endoscopic determination of the lower esophageal sphincter helped place the Bravo probe 5 to 6 cm above the lower esophageal sphincter. A total of 190 studies in 186 patients had sufficiently accurate data. There were 114 women and 72 men with an average age of 51 years. AEAE was defined by a Johnson-DeMeester score greater than 14.7 and was obtained in 115 of 190 studies (61%). Only 64 of 115 patients (56%) demonstrated AEAE for both days of the study, whereas 51 of 115 patients (44%) demonstrated AEAE in a single 24-hour period. There was no difference in the prevalence of AEAE on day 1 versus day 2 only (26% vs. 18%, P = .26). Compared with 24-hour alone data, 48-hour data showed 22% more patients with AEAE. Frequent day-to-day variability in patients with AEAE may be missed by a single 24-hour pH test. Forty-eight-hour pH testing may increase detection accuracy and sensitivity for AEAE by as much as 22%. PMID- 16269376 TI - Quality-of-life after total pancreatectomy: is it really that bad on long-term follow-up? AB - While selected pancreatic diseases may be best treated by total pancreatectomy (TP), the anticipated sequelae of pancreatic insufficiency make TP an undesirable alternative. Our aim was to determine if patients undergoing TP have a worse quality of life (QoL) than age- and gender-matched controls and poor long-term glycemic control. Ninety-nine patients undergoing TP from 1985 through 2002 were identified. The 34 survivors with no recurrent malignancy were surveyed with the Short Form-36 (SF-36), the Audit of Diabetes Dependent QoL (ADD QoL), the European Organization for Research and Treatment in Cancer Pancreas 26 (EORTC PAN 26), and our institutional questionnaire. Operative morbidity and mortality were 32% and 5%, respectively. Three late postoperative deaths (3%) were attributed to hypoglycemia. Of the 34 surviving patients, 27 (79%) agreed to participate at a mean of 7.5 years postoperatively. Seven patients had required 12 hospitalizations for poor glycemic control. Per the SF-36, two domains (role physical and general health) were decreased compared with an age- and gender matched national population (P < .05). The ADD QoL demonstrated an overall decrease in QoL related specifically to the diabetes mellitus (P < .01), but comparison with insulin-dependent diabetics from other causes showed no significant difference in QoL. The EORTC PAN 26 instrument also showed measurable effects on QoL. Total pancreatectomy can be performed safely. QoL after TP is decreased compared with age- and gender-matched controls but not with diabetes from other causes; however, the changes are not overwhelming. TP should remain a viable option but in selected patients. PMID- 16269378 TI - Current practice patterns in pancreatic surgery: results of a multi-institutional analysis of seven large surgical departments in Germany with 1454 pancreatic head resections, 1999 to 2004 (German Advanced Surgical Treatment study group). AB - Despite decreasing mortality rates, morbidity is still high after pancreatic head resection. Comparative data in the United States and Europe show a relationship between hospital volume and mortality. Treatment strategies vary frequently, partially because of the lack of evidence-based data. We performed a multi institutional analysis in Germany evaluating current numbers, indications, techniques, and complication rates of pancreatic head resection. Questionnaires were completed by seven high-volume surgical departments regarding quantitative and qualitative aspects of pancreatic head resections in the period from 1999 to 2004 (five prospective and two retrospective institutional databases). A total of 1454 pancreatic head resections (944 for malignancy) were reported. Mean annual hospital volume ranged from 14 to 52 (10 to 43 in malignancy). Mortality was between 1.1% and 4.8%, morbidity was between 24% and 46%, and pancreatic leakage was between 9% and 20%. In malignant disease, all centers perform standard lymphadenectomy and regard arterial infiltration as a contraindication for resection. However, the rate of portal vein resection varied from 0% to 28%. No consensus is seen on the type of surgery for malignancy and chronic pancreatitis. After resection for pancreatic cancer less than one fourth of the patients receive adjuvant therapy. The results of our analysis in Germany confirm that pancreatic head resection can be performed with low mortality in specialized units. Variations in indications, operative technique, and perioperative care may demonstrate the lack of evidence-based data and/or personal and institutional experience. The low number of patients receiving adjuvant therapy after resection of pancreatic cancer suggests that more efforts must be made to establish novel adjuvant therapies under randomized study conditions. PMID- 16269377 TI - 5-fluorouracil and gemcitabine potentiate the efficacy of oncolytic herpes viral gene therapy in the treatment of pancreatic cancer. AB - Oncolytic herpes viruses are attenuated, replication-competent viruses that selectively infect, replicate within, and lyse cancer cells and are highly efficacious in the treatment of a wide variety of experimental cancers. The current study seeks to define the pharmacologic interactions between chemotherapeutic drugs and the oncolytic herpes viral strain NV1066 in the treatment of pancreatic cancer cell lines. The human pancreatic cancer cell lines Hs 700T, PANC-1, and MIA PaCa-2 were treated in vitro with NV1066 at multiplicities of infection (MOI; ratio of the number of viral particles per tumor cell) ranging from 0.01 to 1.0 with or without 5-fluorouracil (5-FU) or gemcitabine. Synergistic efficacy was determined by the isobologram and combination-index methods of Chou and Talalay. Viral replication was measured using a standard plaque assay. Six days after combination therapy, 76% of Hs 700T cells were killed compared with 43% with NV1066 infection alone (MOI = 0.1) or 0% with 5-FU alone (2 micromol/L) (P < .01). Isobologram and combination-index analyses confirmed a strongly synergistic pharmacologic interaction between the agents at all viral and drug combinations tested (LD5 to LD95) in the three cell lines. Dose reductions up to 6- and 78-fold may be achieved with combination therapy for NV1066 and 5-FU, respectively, without compromising cell kill. 5-FU increased viral replication up to 19-fold compared with cells treated with virus alone. Similar results were observed by combining gemcitabine and NV1066. We have demonstrated that 5-FU and gemcitabine potentiate oncolytic herpes viral replication and cytotoxicity across a range of clinically achievable doses in the treatment of human pancreatic cancer cell lines. The potential clinical implications of this synergistic interaction include improvements in efficacy, treatment-associated toxicity, tolerability of therapeutic regimens, and quality of life. These data provide the cellular basis for the clinical investigation of combined oncolytic herpes virus therapy and chemotherapy in the treatment of pancreatic cancer. PMID- 16269379 TI - Pancreas-sparing duodenectomy is effective management for familial adenomatous polyposis. AB - Duodenal adenocarcinoma remains the leading cause of cancer death in familial adenomatous polyposis patients following colectomy. Stratification based on Spigelman's criteria provides a means for determining therapy. Spigelman stage IV patients have been selected for pancreas-sparing duodenectomy. Twenty-one patients underwent resection between 1992 and 2004, with a mean age of 58 +/- 11 years. The mean time from colectomy to duodenectomy was 27 +/- 13 years. Invasive cancer was found in the distal duodenum in one patient. Operative time averaged 327 +/- 61 minutes with a mean blood loss of 503 +/- 266 ml. There was no mortality, and eight patients (38%) had 14 complications: six (29%) with delayed gastric emptying, four (19%) with biliary/pancreatic anastomotic leak, one with pancreatitis, and one with wound infection. There were two reoperations: one for delayed gastric emptying and one for an early biliary leak. Mean length of stay was 15 +/- 10 days. Two late complications occurred: a stomal ulcer and an intestinal obstruction at 48 and 24 months, respectively. Mean follow-up was 79 months (range, 3-152 months). Two patients developed polyps in the advanced jejunal limb and were endoscopically treated. Pancreas-sparing duodenectomy represents a definitive treatment for advanced duodenal polyposis and can obviate the need for pancreaticoduodenectomy. PMID- 16269380 TI - Pancreaticoduodenectomy after placement of endobiliary metal stents. AB - Contemporary treatment programs for patients with potentially resectable pancreatic cancer often involve preoperative therapy. When the duration of preoperative therapy exceeds 2 months, the risk of plastic endobiliary stent occlusion increases. Metal stents have much better patency but may complicate subsequent pancreaticoduodenectomy (PD). We evaluated rates of perioperative morbidity, mortality, and stent complications in 272 consecutive patients who underwent PD at our institution from May 2001 to November 2004. Of these 272 patients, 29 (11%) underwent PD after placement of a metal stent, 141 underwent PD after placement of a plastic stent, 10 had PD after biliary bypass without stenting, and 92 had PD without any form of biliary decompression. No differences were found between the Metal Stent group and all other patients in median operative time, intraoperative blood loss, or length of hospital stay. No perioperative deaths occurred in the Metal Stent group versus 3 (1.2%) deaths in the other 243 patients. The incidence of major perioperative complications was similar between the two groups, including the rates of pancreatic fistula, intra abdominal abscess, and wound infection. Furthermore, there were no differences in the perioperative morbidity or mortality rates between patients who underwent preoperative biliary decompression with a stent of any kind (metal or plastic) and those patients who underwent no biliary decompression at all. Metal stent related complications occurred in 2 (7%) of 29 patients during a median preoperative interval of 4.1 months; in contrast, 75 (45%) of the 166 patients who had had plastic stents experienced complications, including 98 stent occlusions, during a median preoperative interval of 3.9 months (P < 0.001). We conclude that the use of expandable metal stents does not increase PD-associated perioperative morbidity or mortality, and as such an expandable metal stent is our preferred method of biliary decompression in patients with symptomatic malignant distal bile duct obstruction in whom surgery is not anticipated, or in whom there is a significant delay in the time to surgery. PMID- 16269381 TI - Effects of gastric bypass procedures on bone mineral density, calcium, parathyroid hormone, and vitamin D. AB - Weight loss after gastric bypass procedures has been well studied, but the long term metabolic sequelae are not known. Data on bone mineral density (BMD), calcium, parathyroid hormone, and vitamin D were collected preoperatively and at yearly intervals after gastric bypass procedures. A total of 230 patients underwent preoperative BMD scans. Fifteen patients were osteopenic preoperatively, and three patients subsequently developed osteopenia postoperatively within the first year. No patient had or developed osteoporosis. At 1 year, total forearm BMD decreased by 0.55% (n = 91; P = .03) and radius BMD had increased overall by 1.85% (n = 23; P = .008); both total hip and lumbar spine BMD decreased by 9.27% (n = 22; P < .001) and 4.53% (n = 31; P < .001), respectively. By the second postoperative year, BMD in the total forearm had decreased an additional 3.62% (n = 14; P < .001), whereas radius BMD remained unchanged. Although total hip and lumbar spine BMD significantly decreased at 1 year, by year 2 both total hip and lumbar spine BMD only slightly decreased and were not significantly different from before the operation. Serum calcium decreased from 9.8 mg/dL to 9.2 during the first year (not significant [NS]) and then to 8.8 (NS) by the second year. Parathyroid hormone increased from 59.7 pg/mL (nl 10-65 pg/mL) preoperatively to 63.1 during year 1 (NS) and continued to increase to 64.7 by year 2 (NS). No difference was noted among levels of 25 hydroxy vitamin D preoperatively (25.2 ng/mL; nl 10-65 ng/mL), at 1 year (34.4), and at 2 years (35.4). Our data indicate that bone loss is highest in the first year after gastric bypass with stabilization, and that, in some cases, there is an increase in bone density after the first year. PMID- 16269382 TI - Is Roux-en-Y gastric bypass surgery the most effective treatment for type 2 diabetes mellitus in morbidly obese patients? AB - Type 2 diabetes mellitus (T2DM) has a very strong association with obesity. The aim of our study was to analyze the effects of Roux-en-Y gastric bypass (RYGB) surgery on the glucose metabolism in morbidly obese patients with T2DM. Morbidly obese patients (n = 117) with T2DM underwent measurements of fasting serum glucose and glycosylated hemoglobin (HbA1C) at baseline, 6 months, and 12 months after laparoscopic RYGB surgery. Logistic regression was used in both univariate and multivariate modeling to identify independent variables associated with complete resolution of T2DM. Twelve months after surgery, fasting plasma glucose decreased from a preoperative mean of 164 +/- 55 mg/dL to 101 +/- 38 mg/dL (P = .001) and HbA1C decreased from a preoperative mean of 7.7% +/- 1.5% to 6.0% +/- 1.1% (P = .001). Resolution of T2DM was achieved in 72 patients (74%). All of the remaining 25 patients decreased the daily medication requirements. On univariate analysis, preoperative variables associated with resolution of T2DM were waist circumference, HbA1C, and absence of insulin treatment. Waist circumference (odds ratio 2.4; 95% confidence interval 1.4-4.1; P = .001) and treatment without insulin (odds ratio 42.2; 95% confidence interval 4.3-417.3; P = .002) remained significant predictors of T2DM resolution in the multivariate logistic regression model after adjusting for covariates. Laparoscopic RYGBP resulted in significant resolution of T2DM. Peripheral fat distribution (smaller waist circumference) and absence of insulin treatment were independent and significant predictors of complete resolution of T2DM. PMID- 16269383 TI - Changes in C-reactive protein predict insulin sensitivity in severely obese individuals after weight loss surgery. AB - The production of inflammatory mediators by abdominal adipose tissue may link obesity and insulin resistance. We determined the influence of systemic levels of interleukin-6 and C-reactive protein on insulin sensitivity after weight loss via Roux-en-Y gastric bypass surgery. Severely obese individuals (n = 15) were evaluated at baseline and at 6 months after surgery. Insulin sensitivity was determined by frequently sampled intravenous glucose tolerance testing at the same time points. Visceral and subcutaneous adipose tissue volumes were quantified by computed tomography. Interleukin-6 and C-reactive protein were measured by enzyme-linked immunoassay in plasma and in adipose tissue biopsies. Correlation analysis was used to determine associations between insulin sensitivity and other outcome variables. Significance was set at P < 0.05. Plasma interleukin-6 concentrations were significantly correlated to the IL-6 content of subcutaneous adipose tissue (r = 0.71). At 6 months postsurgery, subcutaneous and visceral adipose tissue volumes were significantly reduced (34.7% and 44.1%, respectively) and insulin sensitivity had improved by 160.9%. Significant longitudinal correlations were found between insulin sensitivity and plasma C reactive protein (r = -0.61), but not plasma interleukin-6 at 6 months. These findings offer insights that link obesity and insulin resistance via the activity of inflammatory mediators. PMID- 16269384 TI - Development of a new access device for transgastric surgery. AB - Flexible endoscope-based endoluminal and transgastric surgery for cholecystectomy, appendectomy, bariatric, and antireflux procedures show promise as a less invasive form of surgery. Current endoscopes and instruments are inadequate to perform such complex surgeries for a variety of reasons: they are too flexible and are insufficient to provide robust grasping and anatomic retraction. The lack of support for a retroflexed endoscope in the peritoneal cavity makes it hard to reach remote structures and makes vigorous retraction of tissues and organs difficult. There is also a need for multiple channels in scopes to allow use of several instruments and to provide traction/countertraction. Finally, secure means of tissue approximation are critical. The aim was to develop and test a new articulating flexible endoscopic system for endoluminal and transgastric endosurgery. A multidisciplinary group of gastrointestinal physicians and surgeons worked with medical device engineers to develop new devices and instruments. Needs assessments and design parameters were developed by consensus. Prototype devices were tested using inanimate models until usable devices were arrived at. The devices were tested in nonsurvival pigs and dogs. The devices were accessed through an incision in the wall of the stomach and manipulated in the peritoneal cavity to accomplish four different tasks: right upper quadrant wedge liver biopsy, right lower quadrant cecal retraction, left lower quadrant running small bowel, and left lower quadrant exposure of esophageal hiatus. In another three pigs, transgastric cholecystectomy was attempted. The positions of the device, camera, and endosurgical instruments, with and without ShapeLock technology, were recorded using laparoscopy and endoscopy and procedure times and success rates were measured. Instrument design parameters and their engineering solutions are described. Flexible multilumen guides which could be locked in position, including a prototype which allowed triangulation, were constructed. Features of the 18-mm devices include multidirectional mid body and/or tip angulation, two 5.5-mm accessory channels allowing the use of large (5-mm) flexible endosurgical instruments, as well as a 4-mm channel for an ultraslim prototype video endoscope (Pentax 4 mm). Using the resulting devices, the four designated transgastric procedures were performed in anesthetized animals. One hundred percent of the transgastric endosurgical procedures were accomplished with the exception of a 50% success for hiatal exposure, a 90% success rate for wedge liver biopsy, and a 33.3% success rate for cholecystectomy. A new endosurgical multilumen device and advanced instrumentation allowed effective transgastric exploration and procedures in the abdominal cavity including retraction of the liver and stomach to allow exposure of the gallbladder, retraction of the cecum, manipulation of the small bowel, and exposure of the esophageal hiatus. This technology may serve as the needed platform for transgastric cholecystectomy, gastric reduction, fundoplication, hiatus hernia repair, or other advanced endosurgical procedures. PMID- 16269385 TI - Virally directed fluorescent imaging improves diagnostic sensitivity in the detection of minimal residual disease after potentially curative cytoreductive surgery. AB - Completeness of cytoreduction is an independent prognostic factor after cure intended surgery for peritoneal carcinomatosis. NV1066, a genetically engineered herpes simplex virus carrying the transgene for green fluorescent protein, selectively infects cancer cells. We sought to determine the feasibility of virally directed fluorescent imaging in the intraoperative detection of minimal residual disease after cytoreductive surgery. NV1066 infected human gastric cancer cells, OCUM-2MD3, and mesothelioma JMN cells at all doses. The infected cells expressed green fluorescent protein and were killed. OCUM-2MD3, and mesothelioma JMN cells at all doses. Peritoneal carcinomatosis was established in mice by injection of OCUM cells into the peritoneal cavity. Forty-eight hours after intraperitoneal injection of NV1066, two experienced surgeons resected all visible disease and identified mice free of disease. Eight of 13 mice thought to be free of disease were found to have residual disease as identified by green fluorescence (mean number of observations: 5; range: 1-9). Residual disease was most frequently observed in the retroperitoneum, pelvis, peritoneal surface, and liver. Specificity of NV1066 infection to tumor nodules was confirmed by immunohistochemistry and by polymerase chain reaction for viral gene. Virally directed fluorescent imaging, a novel molecular imaging technology, can be used for real-time visualization of minimal residual disease after cytoreductive surgery and can improve the completeness of cure-intended resection. PMID- 16269386 TI - Contrast-enhanced intraoperative ultrasonography during hepatectomies for colorectal cancer liver metastases. AB - Preliminary reports showed that contrast-enhanced intraoperative ultrasonography (CEIOUS) provides information on primary or metastatic tumors of the liver that is not obtainable with conventional intraoperative ultrasonography (IOUS). This study validates the impact of CEIOUS, focusing on resective surgery for colorectal cancer (CRC) liver metastases. Twenty-four consecutive patients underwent liver resection using IOUS and CEIOUS for CRC liver metastases. CEIOUS was accomplished with intravenous injection of 4.8 mL of sulphur-hexafluoride microbubbles. CEIOUS found lesions missed at preoperative imaging and at IOUS in four patients and confirmed all of the new findings of IOUS in four patients. In addition, CEIOUS helped to define the tumor margins of the main lesion in 29% of patients with CRC liver metastases. No adverse effects were observed in relation with CEIOUS. In conclusion, CEIOUS improves IOUS accuracy with a significant impact on surgical strategy and radicality in patients who undergo surgery for CRC liver metastases. PMID- 16269387 TI - Molecular absorbent recirculating system for the treatment of acute liver failure in surgical patients. AB - The Molecular Adsorbent Recirculating System (MARS) represents an attractive artificial liver support system for the treatment of liver insufficiency. However, neither indications for MARS treatment (i.e., after extended liver resection) nor criteria for discontinuation of therapy have been evaluated. Therefore, we analyzed the clinical data of all our surgical patients who received MARS treatment for acute liver failure (n = 7). The aim of the study was to identify prognostic indicators for survival. Four of 174 patients resected for hepatic malignancy at our institution received a total of 13 MARS treatments. Two additional patients were successfully bridged to orthotopic liver transplantation with seven MARS treatments and one patient was MARS supported after liver transplantation of a steatotic graft with three MARS treatments. Five of the seven patients survived and were dismissed an average of 31 days, ranging from 17 to 47 days, after the final MARS treatment. No technical complications or adverse effects were observed during the MARS treatments. Important prognostic factors for hepatic recovery and survival were indocyanin green plasma disappearance rates greater than 5%/min and an increase in clotting factor V levels after each MARS treatment. We conclude that MARS therapy can be an effective treatment of postoperative liver insufficiency in the surgical hepatobiliary unit. PMID- 16269388 TI - Incidence and management of biliary leakage after hepaticojejunostomy. AB - This study analyzes the change in the management of biliary leakage after hepaticojejunostomy. Between 1993 and 2003 all patients (n = 1033) were studied with a hepaticojejunostomy as part of a pancreatoduodenectomy (n = 486), proximal bile duct resection (without liver resection) (n = 35), and biliodigestive bypass for malignant (n = 302) and benign (n = 210) disease. Biliary leakage was defined as the presence of bile-stained fluid (>50 mL) in the abdominal drain more than 24 hours after surgery, proven radiologically or at relaparotomy. The studied patients were divided into two equal periods to analyze the change in management. Overall, 24 of 1033 patients (2.3%) had biliary leakage. In multivariate analysis, a body mass index greater than 35 kg/m2 (P = .012), endoscopic biliary drainage (P = .044), and an anastomosis on the segmental bile ducts (P < .001) were independent predictors of leakage. Management in the first half of the study period (1993-1998) versus the second half (1999-2003) was maintenance of operatively placed drains (18% vs. 15%, respectively, P = 1.000), percutaneous transhepatic biliary drainage (18% vs. 69%, respectively, P = .012), surgical drainage (55% vs. 8%, respectively, P = .023), and re-hepaticojejunostomy (9% vs. 8%, respectively, P = 1.000). There was no mortality in the patients with biliary leakage. Leakage after a hepaticojejunostomy is a relatively rare complication without mortality and can safely be managed with percutaneous transhepatic biliary drainage. PMID- 16269389 TI - Laparoscopic versus standard appendectomy outcomes and cost comparisons in the private sector. AB - Minimally invasive surgery has been proposed as the preferred treatment strategy for various gastrointestinal disorders due to shorter hospital stay, less pain, quicker return to normal activities, and improved cosmesis. However, these advantages may not be straightforward for laparoscopic appendectomy, and optimal management of remains controversial. One hundred forty-eight patients with clinical and radiologic diagnoses of acute appendicitis treated in two different hospitals were retrospectively reviewed. Seventy-eight patients underwent laparoscopic appendectomy in hospital A and 70 patients underwent standard appendectomy in hospital B. Patients treated by either type of surgery were compared in terms of clinical and pathologic features, operation characteristics, complications, and costs. There were no significant differences between both groups in terms of clinical features, radiologic studies, complications, and final pathology findings (P > .05). Hospital stay was significantly shorter and bowel movements recovered quicker in the laparoscopy group. However, overall and operating room costs were significantly higher in patients treated by laparoscopy (P < .01). Our series show a subtle difference in terms of hospital stay and bowel movement recovery, favoring patients treated by laparoscopy. However, these results have to be carefully examined and weighed, because overall costs and operating room costs were significantly higher in the laparoscopy group. PMID- 16269390 TI - Adenocarcinomas of the jejunum and ileum: a 25-year experience. AB - Adenocarcinomas of the jejunum and ileum are rare gastrointestinal malignancies. Because few large published experiences exist, we reviewed patients with jejunal and ileal adenocarcinoma treated at our institution over the last 25 years. Between January 1976 and December 2001, 77 patients had an operation for a jejunal or ileal adenocarcinoma. Records were retrospectively reviewed for patient, tumor, and treatment variables. Factors affecting disease recurrence and patient survival were investigated. Fifty-two of the adenocarcinomas (67%) occurred in the jejunum and 25 occurred in the ileum (33%). Mean patient age was 63 +/- 14 years. Segmental bowel resection was performed in 50 patients (65%) with curative intent. Palliative operative procedures including resection or bypass were performed in 27 patients (35%). One (1%) patient had stage I, 18 (23%) stage II, 19 (25%) stage III, and 39 (51%) stage IV adenocarcinoma at diagnosis. Postoperatively, 12 patients had palliative and 18 adjuvant chemotherapy (n = 30), radiation therapy (n = 1), or combination treatment (n = 7). Median patient survival was 19 months. Sixty-six percent of patients who had a curative operation had a tumor relapse. Tumor stage had a highly significant effect (P < 0.0001) on median survival (72 months for stage I and II, 30 months for stage III, and 9 months for stage IV disease). In multivariate analysis of patients having curative treatment, tumor recurrence (P < 0.0001), stage (P < 0.0002), and weight loss (P < 0.001) were significant negative prognostic indicators. Most patients with adenocarcinoma of the jejunum or ileum present with advanced disease. Tumor stage, disease recurrence, and weight loss predicted patient outcome following a curative operation. Early recognition of these tumors requires a high index of suspicion. PMID- 16269391 TI - Hansenula polymorpha Vam7p is required for macropexophagy. AB - We have analyzed the functions of two vacuolar t-SNAREs, Vam3p and Vam7p, in peroxisome degradation in the methylotrophic yeast Hansenula polymorpha. A Hp vam7 mutant was strongly affected in peroxisome degradation by selective macropexophagy as well as non-selective microautophagy. Deletion of Hp-Vam3p function had only a minor effect on peroxisome degradation processes. Both proteins were located at the vacuolar membrane, with Hp-Vam7p also having a partially cytosolic location. Previously, in baker's yeast Vam3p and Vam7p have been demonstrated to be components of a t-SNARE complex essential for vacuole biogenesis. We speculate that the function of this complex in macropexophagy includes a role in membrane fusion processes between the outer membrane layer of sequestered peroxisomes and the vacuolar membrane. Our data suggest that Hp-Vam3p may be functionally redundant in peroxisome degradation. Remarkably, deletion of Hp-VAM7 also significantly affected peroxisome biogenesis and resulted in organelles with multiple, membrane-enclosed compartments. These morphological defects became first visible in cells that were in the mid-exponential growth phase of cultivation on methanol, and were correlated with accumulation of electron-dense extensions that were connected to mitochondria. PMID- 16269392 TI - Protein translocation machineries: how organelles bring in matrix proteins. AB - Eukaryotic cells contain several thousands of proteins that have to be accurately partitioned over the components of the cytoplasm (cytosol or any of the known organelles) to allow proper cell function. To this end, various specific topogenic signals have been designed as well as highly selective protein translocation machineries that ensure that each newly synthesized polypeptide reaches its correct subcellular destination or, in case of secretory proteins, is exported to the cell exterior. This contribution gives an overview regarding the principles of the main examples of polypeptide sorting and translocation, with emphasis on the function of cofactor binding in peroxisomal matrix protein import. PMID- 16269393 TI - Production, purification and characterisation of recombinant Fahsin, a novel antistasin-type proteinase inhibitor. AB - Serine proteinases from inflammatory cells, including polymorphonuclear neutrophils, are involved in various inflammatory disorders, like pulmonary emphysema and rheumatoid arthritis. Inhibitors of these serine proteinases are potential drug candidates for the treatment of these disorders, since they prevent the unrestricted proteolysis. This study describes a novel specific antistasin-type inhibitor of neutrophil serine proteinases, we called Fahsin. This inhibitor was purified from the Nile leech Limnatis nilotica, sequenced and heterologously expressed using a synthetic gene in the methylotrophic yeast Pichia pastoris, yielding 0.5 g(-l) of the protein in the culture medium. Recombinant Fahsin was purified to homogeneity and characterised by N-terminal amino acid sequencing and mass spectrometry. Inhibition-kinetic analysis showed that recombinant Fahsin is a fast, tight-binding inhibitor of human neutrophil elastase with inhibition constant in the nanomolar range. Furthermore, recombinant Fahsin was, in contrast to various other neutrophil elastase inhibitors, insensitive to chemical oxidation and biological oxidation via myeloperoxidase-generated free oxygen radicals. Thus, Fahsin constitutes a novel member of a still expanding family of naturally occurring inhibitors of serine proteinases with potential therapeutic use for treatment of human diseases. PMID- 16269394 TI - Why do we need magnetic resonance imaging in cardiology? PMID- 16269395 TI - Primary PCI for ST elevation AMI save lives and money--what more do we want? PMID- 16269396 TI - Semi-automatic quantification of myocardial infarction from delayed contrast enhanced magnetic resonance imaging. AB - OBJECTIVE: Accurate and reproducible assessment of myocardial infarction is important for treatment planning in patients with ischemic heart disease. This study describes a novel method to quantify myocardial infarction by semi automatic delineation of hyperenhanced myocardium in delayed contrast enhanced (DE) magnetic resonance (MR) images. DESIGN: The proposed method automatically detects the hyperenhanced tissue by first determining the signal intensity of non enhanced myocardium. A fast level set algorithm was used to limit the heterogeneity of the hyperenhanced regions, and to exclude small regions that constitute noise rather than infarction. The method was evaluated in 40 patients; 20 with acute infarction and 20 with chronic healed infarction using scanners from two different manufacturers. Infarct size measured by the proposed semi automatic method was compared with manual measurements from three experienced observers. The software used is freely available for research purposes at http://segment.heiberg.se. RESULTS: The difference in infarct size between semi automatic quantification and the mean of the three observers was 6.1+/-6.6 ml (mean+/-SD), and the interobserver variability (SD) was 4.2 ml. CONCLUSIONS: The method presented is a highly automated method for analyzing myocardial viability from DE-MR images. The bias of the method is acceptable and the variability is in the same order of magnitude as the interobserver variability for manual delineations. PMID- 16269397 TI - Cost-effectiveness of primary percutaneous coronary intervention versus thrombolytic therapy for acute myocardial infarction. AB - OBJECTIVES: We sought to determine the long-term cost-effectiveness of two reperfusion modalities in patients with acute ST-segment elevation myocardial infarction: primary percutaneous coronary intervention (PCI) versus thrombolytic therapy. DESIGN: A state-transition model that follows patients from when they develop STEMI until they die was developed. The model encompassed events and health states. Sensitivity analyses were undertaken. RESULTS: For a 65-year old man, life expectancy was 8.3 years with primary PCI and 7.6 years with thrombolytic therapy. The lifetime costs were 19,250 euros (NOK 154,000) and 29,250 euros (NOK 234,000), respectively, for patients living close to an invasive unit. Cost savings from PCI were mainly due to the reduction in future coronary interventions. For patients needing helicopter transport to arrive in time to an invasive unit for PCI, the costs were 24,000 euros (NOK 192,000) and 29,250 euros (NOK 234,000), respectively (all costs undiscounted). For women, the estimates were somewhat higher due to lower mortality. CONCLUSION: Compared with thrombolytic therapy, reperfusion by primary PCI results in greater health benefits at reduced lifetime costs. These findings may have important clinical implications in an increasing cost-conscious health care environment. PMID- 16269399 TI - Atrial fibrillation in elderly patients after coronary artery bypass grafting; gender differences in outcome. AB - OBJECTIVES: To evaluate gender-related differences in preoperative risk factors, hospital events, especially atrial fibrillation (AF), and length of stay in elderly patients undergoing isolated coronary artery bypass grafting (CABG). DESIGN: Prospectively collected data from consecutive patients undergoing isolated CABG in Tampere University Hospital between May 1999 and November 2000, in total 1131 patients. RESULT: We analysed 621 patients 65 years and older of whom 401 (65%) were male. When evaluating gender differences we found that the women were older (73 vs. 71 yrs, p<0.001) and significantly more often had hypertension (66% vs. 49%, p<0.001) and chronic heart insufficiency (11% vs. 4%, p=0.001). The NYHA classification of the women was worse (3.4 vs. 3.1, p<0.001) and in the angiographic data they had left main stenosis more often (31% vs. 21%, p=0.005) than the men. The women needed longer hospitalisation (19 vs. 15 days, p<0.001). There was no difference in the prevalence of postoperative AF between the genders, but in spite of that women had more postoperative strokes (6% vs. 3%, p=0.028) and also other major complications (29% vs. 19%, p=0.004) than the men. The 30-day mortality was higher in the female group (8% vs. 5%, p=0.06). We analysed preoperative risk factors and found that the females had a 1.6-fold risk for postoperative major complication after adjustment for age and other risk factors. In a logistic regression analysis age and the number of anastomoses emerged as independent predictors of AF in males, but in females we could not find any predictor for postoperative AF. CONCLUSIONS: The women are older and have more comorbidities and risk factors at the time of CABG. They also have more postoperative complications, but not a significantly higher mortality. Importantly, the excess of morbidity remains after adjustment for age and underlying risk factors. There is no gender difference in the incidence of postoperative AF in the elderly population. However, it seems to occur independent of age in the women only. PMID- 16269398 TI - The coronary flow velocity reserve measured by stress transoesophageal echocardiography evaluates the success of coronary interventions--results of a 5 year follow-up. AB - OBJECTIVES: The aim of the present study was to examine the long-term prognostic value of coronary flow velocity reserve (CFR) evaluated by means of stress transoesophageal echocardiography (STEE) in patients who have undergone percutaneous coronary intervention (PCI). DESIGN: The study comprised 31 patients with significant LAD stenosis who underwent LAD-PCI. In consequence of their clinical signs, 11 subjects required rePCI or coronary artery bypass graft (CABG) operation within six months. The clinical status of the remaining 20 cases improved during the follow-up. STEE examinations were performed before LAD-PCI and after it. RESULTS: The CFR of patients in a stable clinical condition improved during the follow-up, while the CFR of those who required rePCI or CABG remained unchanged. From this patient population, two subjects died during the 5 year follow-up. CONCLUSIONS: Most of the patients who displayed an improved CFR after PCI suffered no major clinical events during the 5-year follow-up; in contrast, in those who a priori had a low CFR and did not show any improvement after PCI, major events did occur during this period. PMID- 16269400 TI - Cerebrovascular accidents after cardiac surgery: an analysis of CT scans in relation to clinical symptoms. AB - There is a link between aortic manipulation, particle embolization, and cerebrovascular accidents (CVA) in cardiac surgery. The present aim was to study hemispheric side differences of CVA. Cardiac-surgery patients with CVA and with computer tomography (CT) performed (n = 77) were analyzed within a total group of 2641 consecutive cases. CT data were reviewed for hemispheric and vascular distribution, and compared with CVA-symptom data of immediate and delayed type. Of the included patients, 66% had positive CT. In the group of 'cardiac-type' operations (e.g., routine clamping and cannulation) and having immediate CVA, right-hemispheric lesions were more frequent than of the contra-lateral side (p = 0.005). Patients with aortic dissections had strong dominance of bilateral findings, which was different from the unilateral pattern of 'cardiac-type' operations (p = 0.001). The middle-cerebral artery territory dominated, and when involved showed a significant (p = 0.022) right-sided distribution. Both CT and clinical symptoms confirmed that CVA after cardiac surgery has a right hemispheric predominance. These observations may imply that aortic manipulation directs embolic material towards the brachiocephalic trunk. PMID- 16269401 TI - Postoperative treatment with low molecular weight heparin after right heart assist for coronary artery bypass grafting. AB - Right heart assist (RHA) was used for coronary artery bypass grafting (CABG). We explored the affection of the coagulation system during surgery and evaluated two different antithrombotic treatments postoperatively. The pilot study comprised 14 patients. During surgery activated clotting time (ACT) was kept > 200 sec. By random the patients were selected to different postoperative treatments. The control group received acetyl salicylic acid (ASA) 150 mg daily, the intervention group received ASA 150 mg daily and Low Molecular Weight Heparin (LMWH) 5000 IU x2 for three days. Serum levels of prothrombin fragment 1 and 2 (F 1 + 2), plasmin-antiplasmin product (PAP), anti-Xa activity and functional antithrombin (ATIII) were measured. During surgery there was no increase of F 1 + 2 or PAP. After protamin was administered there was a significant increase of F 1 + 2 but not in PAP during the next 6 hours. Postoperative antithrombotic treatment with LMWH seems to normalise F1 + 2 while ASA does not. ACT level > 200 sec. seems sufficient for RHA-CABG surgery. Fibrinolytic agents are not necessary. It seems that postoperative LMWH treatment prevents increased thrombin formation. General recommendations with respect to antithrombotic treatment beyond ASA can not be made based on study. PMID- 16269402 TI - Activation of beta2-adrenergic receptor plays a pivotal role in generating the protective effect of ischemic preconditioning in rat hearts. AB - BACKGROUND: Ischemic preconditioning (IPC) protects hearts against ischemia by reducing infarct size. However, IPC does not preserve cardiac function, such as left ventricular peak developed pressure (LVPDP). Moreover, IPC fails to protect the post-myocardial infarct (MI) heart. DESIGN: Rat hearts were transfected with beta2-adrenergic receptor (B2AR) cDNA by the hemagglutinating virus of Japan liposome method. After the gene transfer, the hearts were perfused in a Langendorff mode and preconditioned with two cycles of 5 min of ischemia and reperfusion. After 20 min of global ischemia, the hearts were reperfused under aerobic conditions for 90 min. LVPDP was measured as an indicator of the cardiac function. RESULTS: LVPDP of ischemic hearts was well preserved by the combination treatment of IPC and gene transfer of B2AR, but not IPC or gene transfer of B2AR alone. Moreover, the treatment was beneficial to even the post-MI heart. On the contrary, gene transfer of beta-adrenergic receptor kinase 1 (BARK1) reduced the protective effect of IPC. We also found that the mRNA ratio of B2AR and BARK1 was well correlated with the preservation of the LVPDP. CONCLUSIONS: The combination treatment of IPC and gene transfer of B2AR protects cardiac function against ischemia and it shows the beneficial effect also in post-MI hearts. PMID- 16269405 TI - Assembly of the Bi-component leukocidin pore examined by truncation mutagenesis. AB - Staphylococcal leukocidin (Luk) and alpha-hemolysin (alphaHL) are members of the same family of beta barrel pore-forming toxins (betaPFTs). Although the alphaHL pore is a homoheptamer, the Luk pore is formed by the co-assembly of four copies each of the two distantly related polypeptides, LukF and LukS, to form an octamer. Here, we examine N- and C-terminal truncation mutants of LukF and LukS. LukF subunits missing up to nineteen N-terminal amino acids are capable of producing stable, functional hetero-oligomers with WT LukS. LukS subunits missing up to fourteen N-terminal amino acids perform similarly in combination with WT LukF. Further, the simultaneous truncation of both LukF and LukS is tolerated. Both Luk subunits are vulnerable to short deletions at the C terminus. Interestingly, the N terminus of the LukS polypeptide becomes resistant to proteolytic digestion in the fully assembled Luk pore while the N terminus of LukF remains in an exposed conformation. The results from this work and related experiments on alphaHL suggest that, although the N termini of betaPFTs may undergo reorganization during assembly, they are dispensable for the formation of functional pores. PMID- 16269404 TI - Glycosylphosphatidylinositol-anchored proteases of Candida albicans target proteins necessary for both cellular processes and host-pathogen interactions. AB - Intracellular and secreted proteases fulfill multiple functions in microorganisms. In pathogenic microorganisms extracellular proteases may be adapted to interactions with host cells. Here we describe two cell surface associated aspartic proteases, Sap9 and Sap10, which have structural similarities to yapsins of Saccharomyces cerevisiae and are produced by the human pathogenic yeast Candida albicans. Sap9 and Sap10 are glycosylphosphatidylinositol-anchored and located in the cell membrane or the cell wall. Both proteases are glycosylated, cleave at dibasic or basic processing sites similar to yapsins and Kex2-like proteases, and have functions in cell surface integrity and cell separation during budding. Overexpression of SAP9 in mutants lacking KEX2 or SAP10, or of SAP10 in mutants lacking KEX2 or SAP9, only partially restored these phenotypes, suggesting distinct target proteins of fungal origin for each of the three proteases. In addition, deletion of SAP9 and SAP10 modified the adhesion properties of C. albicans to epithelial cells and caused attenuated epithelial cell damage during experimental oral infection suggesting a unique role for these proteases in both cellular processes and host-pathogen interactions. PMID- 16269403 TI - Identification and characterization of novel IGFBP5 interacting protein: evidence IGFBP5-IP is a potential regulator of osteoblast cell proliferation. AB - Insulin-like growth factor binding protein-5 (IGFBP5) is a multifunctional protein, which acts not only as a traditional binding protein, but also functions as a growth factor independent of IGFs to stimulate bone formation. It has been predicted that the intrinsic growth factor action of IGFBP5 involves binding of IGFBP5 to a putative receptor to induce downstream signaling pathways and/or nuclear translocation of IGFBP5 to influence transcription of genes involved in osteoblast cell proliferation/differentiation. Our study indentified proteins that bound to IGFBP5 using IGFBP5 as bait in a yeast two-hybrid screen of the U2 human osteosarcoma cell cDNA library. One of the clones that interacted strongly with the bait under high-stringency conditions corresponded to a novel IGFBP5 interacting protein (IGFBP5-IP) encoded by a gene that resides in mouse chromosome 10. The interaction between IGFBP5-IP and IGFBP5 is confirmed by in vitro coimmunoprecipitation studies that used pFlag and IGFBP5 polyclonal antibody, and cell lysates overexpressing both IGFBP5-IP and IGFBP5. Northern blot and RT-PCR analysis showed that the IGFBP-IP is expressed in both untransformed normal human osteoblasts and in osteosarcoma cell lines, which are known to produce IGFBP5. To determine the roles of IGFBP5-IP, we evaluated the effect of blocking the expression of IGFBP5-IP on osteoblast proliferation. We found that using a IGFBP5-IP-specific small interfering-hairpin plasmid resulted in a decrease in both basal and IGFBP5-induced osteoblast cell proliferation. On the basis of these findings, we predict that IGFBP5-IP may act as intracellular mediator of growth promoting actions of IGFBP5 and perhaps other osteoregulatory agents in bone cells. PMID- 16269406 TI - C to U editing stimulates A to I editing in the anticodon loop of a cytoplasmic threonyl tRNA in Trypanosoma brucei. AB - Editing of tRNAs is widespread in nature and either changes the decoding properties or restores the folding of a tRNA. Unlike the phylogenetically disperse adenosine (A) to inosine (I) editing, cytosine (C) to uridine (U) editing has only been previously described in organellar tRNAs. We have shown that cytoplasmic tRNA(Thr)(AGU) undergoes two distinct editing events in the anticodon loop: C to U and A to I. In vivo, every inosine-containing tRNA(Thr) is also C to U edited at position 32. In vitro, C to U editing stimulates conversion of A to I at the wobble base. Although the in vivo and in vitro requirements differ, in both cases, the C to U change plays a key role in A to I editing. Due to an unusual abundance of A34-containing tRNAs, our results also suggest that the unedited and edited tRNAs are functional, each dedicated to decoding a specific threonine codon. C to U editing of cytoplasmic tRNA expands the editing repertoire in eukaryotic cells, and when coupled to A to I changes, leads to an interrelation between editing sites. PMID- 16269407 TI - Role of the transmembrane domain of FXYD7 in structural and functional interactions with Na,K-ATPase. AB - Members of the FXYD family are tissue-specific regulators of the Na,K-ATPase. Here, we have investigated the contribution of amino acids in the transmembrane (TM) domain of FXYD7 to the interaction with Na,K-ATPase. Twenty amino acids of the TM domain were replaced individually by tryptophan, and combined mutations and alanine insertion mutants were constructed. Wild type and mutant FXYD7 were expressed in Xenopus oocytes with Na,K-ATPase. Mutational effects on the stable association with Na,K-ATPase and on the functional regulation of Na,K-ATPase were determined by co-immunoprecipitation and two-electrode voltage clamp techniques, respectively. Most residues important for the structural and functional interaction of FXYD7 are clustered in a face of the TM helix containing the two conserved glycine residues, but others are scattered over two-thirds of the FXYD TM helix. Ile-35, Ile-43, and Ile-44 are only involved in the stable association with Na,K-ATPase. Glu-26, Met-30, and Ile-44 are important for the functional effect and/or the efficient association of FXYD7 with Na,K-ATPase, consistent with the prediction that these amino acids contact TM domain 9 of the alpha subunit (Li, C., Grosdidier, A., Crambert, G., Horisberger, J.-D., Michielin, O., and Geering, K. (2004) J. Biol. Chem. 279, 38895-38902). Several amino acids that are not implicated in the efficient association of FXYD7 with the Na,K-ATPase are specifically involved in the functional effect of FXYD7. Leu-32 and Phe-37 influence the apparent affinity for external K+, whereas Val-28 and Ile-42 are implicated in the apparent affinity for both external K+ and external Na+. These amino acids act in a synergistic way. These results highlight the important structural and functional role of the TM domain of FXYD7 and delineate the determinants that mediate the complex interactions of FXYD7 with Na,K-ATPase. PMID- 16269408 TI - Deficiency of disulfide bonds facilitating fibrillogenesis of endostatin. AB - Endostatin is an endogenous inhibitor of tumor angiogenesis and tumor growth. It has two pairs of disulfide bonds in a unique nested pattern, which play a key role in its native conformation, stability, and activity. Here, we constructed a disulfide-deficient variant of endostatin, endo-all-Ala, to examine the effects of the two disulfide bonds on fibrillogenesis of endostatin under nondenaturing conditions. Based on thioflavin T fluorescence, atomic force microscopy, far-UV circular dichroism, and Fourier transform infrared spectroscopy, we found that endo-all-Ala, which has a higher alpha-helical content compared with wild type, is prone to forming fibrils in a pH-dependent manner. Subsequently, more hydrophobic patches with a lower stability of endo-all-Ala were observed when compared with wild type, which possibly contributes to the propensity of amyloid formation of endo-all-Ala. To our surprise, the significant increase of the alpha helical content in endostatin induced by trifluoroethanol can also facilitate fibril formation. In addition, the cytotoxicity of fibrillar aggregates of endo all-Ala, which were generated at different stages of the fibril formation process, was evaluated by cell viability assay. The results indicate that the cytotoxicity is not due to the fibrils but rather due to the granular aggregates of endo-all-Ala. Moreover, endostatin was interestingly found to be reduced by glutathione at physiological concentrations. Our present work not only elucidates the correlation between the existence of disulfide bonds and the fibril formation of endostatin but also may provide some insights into the structural and functional basis of endostatin in Alzheimer disease brains. PMID- 16269409 TI - Essential role of aralar in the transduction of small Ca2+ signals to neuronal mitochondria. AB - Aralar, the neuronal Ca(2+)-binding mitochondrial aspartate-glutamate carrier, has Ca(2+) binding domains facing the extramitochondrial space and functions in the malate-aspartate NADH shuttle (MAS). Here we showed that MAS activity in brain mitochondria is stimulated by extramitochondrial Ca(2+) with an S(0.5) of 324 nM. By employing primary neuronal cultures from control and aralar-deficient mice and NAD(P)H imaging with two-photon excitation microscopy, we showed that lactate utilization involves a substantial transfer of NAD(P)H to mitochondria in control but not aralar-deficient neurons, in agreement with the lack of MAS activity associated with aralar deficiency. The increase in mitochondrial NAD(P)H was greatly potentiated by large [Ca(2+)](i) signals both in control and aralar deficient neurons, showing that these large signals activate the Ca(2+) uniporter and mitochondrial dehydrogenases but not MAS activity. On the other hand, small [Ca(2+)](i) signals potentiate the increase in mitochondrial NAD(P)H only in control but not in aralar-deficient neurons. We concluded that neuronal MAS activity is selectively activated by small Ca(2+) signals that fall below the activation range of the Ca(2+) uniporter and plays an essential role in mitochondrial Ca(2+) signaling. PMID- 16269410 TI - The P2X7 nucleotide receptor mediates skeletal mechanotransduction. AB - The P2X7 nucleotide receptor (P2X7R) is an ATP-gated ion channel expressed in many cell types including osteoblasts and osteocytes. Mice with a null mutation of P2X7R have osteopenia in load bearing bones, suggesting that the P2X7R may be involved in the skeletal response to mechanical loading. We found the skeletal sensitivity to mechanical loading was reduced by up to 73% in P2X7R null (knock out (KO)) mice. Release of ATP in the primary calvarial osteoblasts occurred within 1 min of onset of fluid shear stress (FSS). After 30 min of FSS, P2X7R mediated pore formation was observed in wild type (WT) cells but not in KO cells. FSS increased prostaglandin (PG) E2 release in WT cells but did not alter PGE2 release in KO cells. Studies using MC3T3-E1 osteoblasts and MLO-Y4 osteocytes confirmed that PGE2 release was suppressed by P2X7R blockade, whereas the P2X7R agonist BzATP enhanced PGE2 release. We conclude that ATP signaling through P2X7R is necessary for mechanically induced release of prostaglandins by bone cells and subsequent osteogenesis. PMID- 16269411 TI - Staphylococcus aureus Sortase A transpeptidase. Calcium promotes sorting signal binding by altering the mobility and structure of an active site loop. AB - Many virulence factors in gram-positive bacteria are covalently anchored to the cell-wall peptidoglycan by sortase enzymes, a group of widely distributed cysteine transpeptidases. The Staphylococcus aureus Sortase A protein (SrtA) is the archetypal member of the Sortase family and is activated by Ca2+, an adaptation that may facilitate host colonization as elevated concentrations of this ion are encountered in human tissue. Here we show that a single Ca2+ ion bound to an ordered pocket on SrtA allosterically activates catalysis by modulating both the structure and dynamics of a large active site loop. Detailed nitrogen-15 relaxation measurements indicate that Ca2+ may facilitate the adaptive recognition of the substrate by inducing slow micro- to millisecond time scale dynamics in the active site. Interestingly, relaxation compensated Carr Purcell-Meiboom-Gill experiments suggest that the time scale of these motions is directly correlated with ion binding. The results of site-directed mutagenesis indicate that this motional coupling is mediated by the side chain of Glu-171, which is positioned within the beta6/beta7 loop and shown to contribute to Ca2+ binding. The available structural and dynamics data are compatible with a loop closure model of Ca2+ activation, in which the beta6/beta7 loop fluctuates between a binding competent closed form that is stabilized by Ca2+, and an open, highly flexible state that removes key substrate contacting residues from the active site. PMID- 16269412 TI - Targeting perlecan in human keratinocytes reveals novel roles for perlecan in epidermal formation. AB - Heparin-binding growth factors are crucial for the formation of human epidermis, but little is known about the role of heparan sulfate proteoglycans in this process. Here we investigated the role of the heparan sulfate proteoglycan, perlecan, in the formation of human epidermis, by utilizing in vitro engineered human skin. By disrupting perlecan expression either in the dermis or the epidermis, we found that epidermally derived perlecan is essential for epidermal formation. Perlecan-deficient keratinocytes formed a strikingly thin and poorly organized epidermis because of premature apoptosis and failure to complete their stratification program. Exogenous perlecan fully restored epidermal formation. Perlecan deposition in the basement membrane zone correlated with formation of multilayered epidermis. Perlecan deficiency, however, had no effect on the lining and deposition of major basement membrane components as was evident by a continuous linear staining of laminin and collagen IV. Similarly, perlecan deficiency did not affect the distribution of beta1 integrin. Addition of the perlecan ligand, fibroblast growth factor 7, protected perlecan-deficient keratinocytes from cell death and improved the thickness of the epidermis. Taken together, our results revealed novel roles for perlecan in epidermal formation. Perlecan regulates both the survival and terminal differentiation steps of keratinocytes. Our results suggested a model whereby perlecan regulates these processes via controlling the bioavailability of perlecan-binding soluble factors involved in epidermal morphogenesis. PMID- 16269415 TI - LogoBar: bar graph visualization of protein logos with gaps. AB - SUMMARY: LogoBar is a Java application to display protein sequence logos. In our software gaps are accounted for when calculating the information content present at each residue position in a multiple alignment. The resulting logo is displayed as a graph consisting of bars, although traditional letter representation is also possible. Amino acids are displayed from the bottom up with decreasing frequencies i.e. the most abundant residue is placed at the bottom of the logo. The bars can be color-coded according to user specifications. Gaps in the alignment are also displayed, either on top or at the bottom of the logo. Furthermore, residues can either be arranged according to their relative abundance or grouped according to user criteria to emphasize the conserved nature of particular positions. AVAILABILITY: LogoBar and further documentation is available at http://www.biosci.ki.se/groups/tbu/logobar/ PMID- 16269413 TI - GLUT4 distribution between the plasma membrane and the intracellular compartments is maintained by an insulin-modulated bipartite dynamic mechanism. AB - The GLUT4 glucose transporter is predominantly retained inside basal fat and muscle cells, and it is rapidly recruited to the plasma membrane with insulin stimulation. There is controversy regarding the mechanism of basal GLUT4 retention. One model is that GLUT4 retention is dynamic, based on slow exocytosis and rapid internalization of the entire pool of GLUT4 (Karylowski, O., Zeigerer, A., Cohen, A., and McGraw, T. E. (2004) Mol. Biol. Cell 15, 870-882). In this model, insulin increases GLUT4 in the plasma membrane by modulating GLUT4 exocytosis and endocytosis. The second model is that GLUT4 retention is static, with approximately 90% of GLUT4 stored in compartments that are not in equilibrium with the cell surface in basal conditions (Govers, R., Coster, A. C., and James, D. E. (2004) Mol. Cell Biol. 24, 6456-6466). In this model, insulin increases GLUT4 in the plasma membrane by releasing it from the static storage compartment. Here we show that under all experimental conditions examined, basal GLUT4 retention is by a bipartite dynamic mechanism involving slow efflux and rapid internalization. To establish that the dynamic model developed in studies of the extreme conditions of >100 nm insulin and no insulin also describes GLUT4 behavior at more physiological insulin concentrations, we characterized GLUT4 trafficking in 0.5 nm insulin. This submaximal insulin concentration promotes an intermediate effect on both GLUT4 exocytosis and endocytosis, resulting in an intermediate degree of redistribution to the plasma membrane. These data establish that changes in the steady-state surface/total distributions of GLUT4 are the result of gradated, insulin-induced changes in GLUT4 exocytosis and endocytosis rates. PMID- 16269416 TI - COMPAM :visualization of combining pairwise alignments for multiple genomes. AB - COMPAM is a tool for visualizing relationships among multiple whole genomes by combining all pairwise genome alignments. It displays shared conserved regions (blocks) and where these blocks occur (edges) as block relation graphs which can be explored interactively. An unannotated genome, e.g. can then be explored using information from well-annotated genomes, COG-based genome annotation and genes. COMPAM can run either as a stand-alone application or through an applet that is provided as service to PLATCOM, a toolset for whole genome comparative analysis, where a wide variety of genomes can be easily selected. Features provided by COMPAM include the ability to export genome relationship information into file formats that can be used by other existing tools. AVAILABILITY: http://bio.informatics.indiana.edu/projects/compam/ PMID- 16269414 TI - An efficient comprehensive search algorithm for tagSNP selection using linkage disequilibrium criteria. AB - MOTIVATION: Selecting SNP markers for genome-wide association studies is an important and challenging task. The goal is to minimize the number of markers selected for genotyping in a particular platform and therefore reduce genotyping cost while simultaneously maximizing the information content provided by selected markers. RESULTS: We devised an improved algorithm for tagSNP selection using the pairwise r(2) criterion. We first break down large marker sets into disjoint pieces, where more exhaustive searches can replace the greedy algorithm for tagSNP selection. These exhaustive searches lead to smaller tagSNP sets being generated. In addition, our method evaluates multiple solutions that are equivalent according to the linkage disequilibrium criteria to accommodate additional constraints. Its performance was assessed using HapMap data. AVAILABILITY: A computer program named FESTA has been developed based on this algorithm. The program is freely available and can be downloaded at http://www.sph.umich.edu/csg/qin/FESTA/ PMID- 16269417 TI - PRODISTIN Web Site: a tool for the functional classification of proteins from interaction networks. AB - The PRODISTIN Web Site is a web service allowing users to functionally classify genes/proteins from any type of interaction network. The resulting computation provides a classification tree in which (1) genes/proteins are clustered according to the identity of their interaction partners and (2) functional classes are delineated in the tree using the Biological Process Gene Ontology annotations. AVAILABILITY: The PRODISTIN Web Site is freely accessible at http://gin.univ-mrs.fr/webdistin PMID- 16269418 TI - ClaNC: point-and-click software for classifying microarrays to nearest centroids. AB - SUMMARY: ClaNC (classification to nearest centroids) is a simple and an accurate method for classifying microarrays. This document introduces a point-and-click interface to the ClaNC methodology. The software is available as an R package. AVAILABILITY: ClaNC is freely available from http://students.washington.edu/adabney/clanc PMID- 16269420 TI - Right bundle branch block during the acute phase of myocardial infarction: modern redefinitions of old concepts. PMID- 16269419 TI - Prognostic differences between different types of bundle branch block during the early phase of acute myocardial infarction: insights from the Hirulog and Early Reperfusion or Occlusion (HERO)-2 trial. AB - AIMS: Bundle branch block (BBB) early during acute myocardial infarction (AMI) is often considered high risk for mortality. Little is known about how different BBB types influence prognosis. METHODS AND RESULTS: The HERO-2 trial recruited 17 073 patients with ischaemic symptoms lasting >30 min and either ST elevation with or without right bundle branch block (RBBB) or presumed-new left bundle branch block (LBBB). Electrocardiograms were performed before and 60 min after the start of fibrinolytic therapy. Using patients with normal intraventricular conduction as a reference, odds ratios (ORs) for 30-day mortality were calculated for different BBB types (LBBB, RBBB with anterior AMI, and RBBB with inferior AMI) present at randomization and/or 60 min, with adjustment for recruitment region, pre infarction characteristics, time to randomization, hemodynamics, and Killip class. At randomization, the 873 patients (5.11%) with BBB had worse baseline characteristics than patients without BBB. In patients presenting with LBBB (n=300), the ORs for 30-day mortality were 1.90 (95% CI 1.39-2.59) before and 0.68 (0.48-0.99) after adjustment for other prognosticators. In patients presenting with RBBB (n=415) and anterior AMI, the ORs were 3.52 (2.82-4.38) before and 2.48 (1.93-3.19) after adjustment. In patients presenting with RBBB and inferior AMI (n=158), the ORs were 1.74 (1.06-2.86) before and 1.22 (0.71 2.08) after adjustment. Within 60 min, 143 patients (0.92%) developed new BBB. The adjusted ORs for 30-day mortality were 2.97 (1.16-7.57) in the 25 patients with new LBBB, 3.84 (2.38-6.22) in the 100 with new RBBB and anterior AMI, and 2.23 (0.54-9.21) in the 18 with new RBBB and inferior AMI. CONCLUSION: RBBB accompanying anterior AMI at presentation and new BBB (including LBBB) early after fibrinolytic therapy are independent predictors of high 30-day mortality. These electrocardiographic features should be considered in risk stratification to identify high-risk patients. PMID- 16269421 TI - Signal maps for mass spectrometry-based comparative proteomics. AB - Mass spectrometry-based proteomic experiments, in combination with liquid chromatography-based separation, can be used to compare complex biological samples across multiple conditions. These comparisons are usually performed on the level of protein lists generated from individual experiments. Unfortunately given the current technologies, these lists typically cover only a small fraction of the total protein content, making global comparisons extremely limited. Recently approaches have been suggested that are built on the comparison of computationally built feature lists instead of protein identifications. Although these approaches promise to capture a bigger spectrum of the proteins present in a complex mixture, their success is strongly dependent on the correctness of the identified features and the aligned retention times of these features across multiple experiments. In this experimental-computational study, we went one step further and performed the comparisons directly on the signal level. First signal maps were constructed that associate the experimental signals across multiple experiments. Then a feature detection algorithm used this integrated information to identify those features that are discriminating or common across multiple experiments. At the core of our approach is a score function that faithfully recognizes mass spectra from similar peptide mixtures and an algorithm that produces an optimal alignment (time warping) of the liquid chromatography experiments on the basis of raw MS signal, making minimal assumptions on the underlying data. We provide experimental evidence that suggests uniqueness and correctness of the resulting signal maps even on low accuracy mass spectrometers. These maps can be used for a variety of proteomic analyses. Here we illustrate the use of signal maps for the discovery of diagnostic biomarkers. An imple mentation of our algorithm is available on our Web server. PMID- 16269422 TI - Proteins exposed at the adult schistosome surface revealed by biotinylation. AB - The human blood-dwelling parasite Schistosoma mansoni can survive in the hostile host environment for decades and must therefore display effective strategies to evade the host immune responses. The surface of the adult worm is covered by a living syncytial layer, the tegument, bounded by a complex multilaminate surface. This comprises a normal plasma membrane overlain by a secreted bilayer, the membranocalyx. Recent proteomic studies have identified constituents of the tegument, but their relative locations remain to be established. We labeled the most exposed surface proteins using two impermeant biotinylation reagents that differed only in length. We anticipated that the two reagents would display distinct powers of penetration, thereby producing a differential labeling pattern. The labeled proteins were recovered by streptavidin affinity and identified by tandem mass spectrometry. A total of 28 proteins was identified, 13 labeled by a long form reagent and the same 13 plus a further 15 labeled by a short form reagent. The parasite proteins included membrane enzymes, transporters, and structural proteins. The short form reagent additionally labeled some cytosolic and cytoskeletal proteins, the latter being constituents of the intracellular spines. Only a single secreted protein was labeled, implying a location between the plasma membrane and the membranocalyx or as part of the latter. Four host proteins, three immunoglobulin heavy chains and C3c/C3dg, a fragment of complement C3, were labeled by both reagents indicating their exposed situation. The presence of the degraded complement C3 implicates inhibition of the classical pathway as a major element of the immune evasion strategy, whereas the recovery of only one truly secreted protein points to the membranocalyx acting primarily as an inert protective barrier between the immune system and the tegument plasma membrane. Collectively the labeled parasite proteins merit investigation as potential vaccine candidates. PMID- 16269423 TI - Anti-inflammatory effects of leflunomide in combination with methotrexate on co culture of T lymphocytes and synovial macrophages from rheumatoid arthritis patients. AB - OBJECTIVES: To investigate the anti-inflammatory effects of the active leflunomide metabolite A771726 (Lef-M) in combination with methotrexate (MTX) on synovial macrophages (SM) from rheumatoid arthritis (RA) patients co-cultured with an activated T cell line (Jurkat cell line). METHODS: Pro-inflammatory cytokines (TNFalpha, IL1beta, IL6), adhesion molecule ICAM-1, cyclooxygenase isoenzymes (COX1, COX2), and the nuclear factor kappaB (NF-kappaB) complex were analysed on SM co-cultured with a T cell line, as intracellular protein expression by immunocytochemistry (ICC) and western blot analysis, as extracellular protein expression by ELISA assay, and as mRNA expression by reverse transcriptase-multiplex PCR (RT-MPCR) after treatment with Lef-M (1, 10, 30 micromol/l) alone or in combination with MTX (50 ng/ml). RESULTS: The most significant intracellular decrease in cytokines was observed by ICC in SM treated with the combination of Lef-M (1, 10, 30 micromol/l) and MTX (50 ng/ml) versus untreated SM (TNFalpha 29%, 37%, 49%, IL1beta 56%, 43%, 50%, and IL6 59%, 62%, 71%, respectively). Furthermore, a significant decrease was confirmed concerning cytokine levels evaluated by ELISA in the medium of SM treated with the combination Lef-M+MTX (TNFalpha 40%, 41%, 44%; IL1beta 10%, 20%, 60%; IL6 37%, 41%, 49%, respectively). Western blot and RT-PCR analysis confirmed these results. Concordant decreased expression was observed for ICAM-1, COX1, COX2, and the NF-kappaB complex after Lef-M+MTX treatment. CONCLUSIONS: The combination of MTX and Lef-M shows additive inhibitory effects on the production of inflammatory mediators from SM co-cultured with a T cell line. These observations might support the positive results obtained in RA clinical studies by combination therapy. PMID- 16269424 TI - Repeated B cell depletion in treatment of refractory systemic lupus erythematosus. AB - OBJECTIVES: To report the clinical outcome and safety profile of repeated B cell depletion in seven patients with refractory systemic lupus erythematosus (SLE). METHODS: Since June 2000, seven patients with refractory SLE had repeated cycles of B cell depletion (18 cycles in total, up to three cycles per patient) because of disease relapse. The clinical response (assessed by the British Isles Lupus Activity Guide (BILAG) activity index), duration of B cell depletion, and adverse events in these patients was reviewed. RESULTS: Four patients (Nos 1, 2, 3, 6) had three cycles of treatment and three (Nos 4, 5, 7) had two cycles. Four of the seven patients (Nos 1, 3, 5, 6) improved. The mean global BILAG scores dropped from 15 to 6 at 5-7 months. The median duration of clinical response and B cell depletion was 13 months and 6 months, respectively. After the third cycle, 2/4 patients (Nos 1 and 2) improved. The median duration of clinical benefit was 12 months. Most patients tolerate re-treatment very well. CONCLUSION: Re-treatment with B cell depletion of patients with severe SLE is safe and may be effective for 6-12 months on average. PMID- 16269425 TI - Lack of efficacy of rituximab in Wegener's granulomatosis with refractory granulomatous manifestations. AB - OBJECTIVE: To investigate the safety and efficacy of rituximab (RTX) in patients with refractory Wegener's granulomatosis (WG). PATIENTS AND METHODS: Eight consecutive patients with active refractory WG were included. In all patients disease activity had persisted despite standard treatment with cyclophosphamide and prednisolone, as well as tumour necrosis factor alpha blockade 3 months before inclusion in the study. Patients had particular granulomatous manifestations like retro-orbital granulomata (n=5), nodules of the lungs (n=1), and subglottic stenosis (n=2). RTX was given intravenously every 4th week in combination with the standard treatment in five patients and with methotrexate in two others. Disease extent and activity were monitored clinically by interdisciplinary care, immunodiagnostics (ANCA serology, B cells by flow cytometry), and magnetic resonance imaging. RESULTS: Beneficial response and a reduction in disease activity were seen in three patients, two of whom went into complete remission. In three other patients, disease activity remained unchanged while the disease progressed in the remaining two patients. In all patients peripheral blood B cells fell to zero during treatment with RTX. cANCA titres remained unchanged in all except one patient. CONCLUSION: In this pilot study, B lymphocyte depletion was not associated with a change of the ANCA titres or obvious clinical improvement of refractory granulomatous disease in patients with WG. Further studies are needed to evaluate the role of RTX in WG. PMID- 16269426 TI - Differential influence of p38 mitogen activated protein kinase (MAPK) inhibition on acute phase protein synthesis in human hepatoma cell lines. AB - BACKGROUND: Inhibition of intracellular signal transduction is considered to be an interesting target for treatment in inflammation. p38 MAPK inhibitors, especially, have been developed and are now in phase II clinical trials for rheumatoid arthritis (RA). OBJECTIVE: To investigate the influence of p38 MAPK inhibition on acute phase protein (APP) production, which is dependent on both JAK/STAT and p38 MAPK pathways. METHODS: The effects of p38 MAPK inhibition on APP production and mRNA expression in four human hepatoma cell lines was investigated, after stimulation with interleukin (IL)6 and/or IL1beta or tumour necrosis factor alpha. RESULTS: Two out of four cell lines produced C reactive protein (CRP), especially after combined IL6 and IL1beta stimulation. CRP production was significantly inhibited by the p38 MAPK specific inhibitor RWJ 67657 at 1 micromol/l, which is pharmacologically relevant. Fibrinogen production was also inhibited at 1 micromol/l in all cell lines. Serum amyloid A (SAA) was produced in all four lines. In contrast with CRP, SAA production was not inhibited by RWJ 67657 at 1 micromol/l. CONCLUSION: Production and mRNA expression of CRP and fibrinogen, but not SAA production and mRNA expression, were significantly inhibited by p38 MAPK specific inhibitor in hepatoma cell lines. For p38 MAPK inhibitor treatment in RA SAA might be a better marker of disease activity than CRP and fibrinogen, because SAA is not directly affected by p38 MAPK inhibition. PMID- 16269427 TI - High prevalence of asymptomatic cervical spine subluxation in patients with rheumatoid arthritis waiting for orthopaedic surgery. AB - OBJECTIVE: To study the prevalence of cervical spine subluxation in patients with rheumatoid arthritis waiting for orthopaedic surgery, and symptoms that might be associated with the disorders. METHODS: 194 patients with rheumatoid arthritis were referred for orthopaedic surgery at Jyvaskyla Central Hospital, 154 (79%) of whom volunteered for the present study including clinical examination, laboratory tests, radiographs of the cervical spine, hands, and feet, and self report questionnaires. Definition of anterior atlantoaxial subluxation (aAAS) was >3 mm and of subaxial subluxation (SAS)>or=3 mm. Atlantoaxial impaction (AAI) was analysed following to the Sakaguchi-Kauppi method. RESULTS: 67 patients (44%) had cervical spine subluxation or previous surgical fusion. The prevalence of aAAS, AAI, SAS, or previous fusion was 27 (18%), 24 (16%), 29 (19%), and 8 (5%), respectively; 69% of patients with cervical spine subluxations (those with fusions excluded) reported neck pain, compared with 65% of patients without subluxations (p=0.71). The prevalence of occipital, temporal, retro-orbital, and radicular pain in upper extremities was similar in patients with or without cervical spine subluxations (54% v 43%; 17% v 31%; 25% v 24%; 47% v 48%, respectively). However, patients with subluxations were older, had longer disease duration, more active disease, poorer function according to the Health Assessment Questionnaire, and had more often erosive disease. CONCLUSIONS: Asymptomatic cervical spine subluxation is common in patients with rheumatoid arthritis waiting for orthopaedic surgery. Regardless of symptoms, the possibility of cervical spine subluxation in patients with severe rheumatoid arthritis should be considered in preoperative evaluation. PMID- 16269428 TI - Greater reduction of knee than hip pain in osteoarthritis treated with naproxen, as evaluated by WOMAC and SF-36. AB - OBJECTIVES: To compare the improvement of hip and knee osteoarthritis during treatment with naproxen. METHODS: Men and women aged 40 to 75 years with symptomatic osteoarthritis of the knee or hip of at least three months' duration participated in a six week placebo controlled, double blind study with naproxen 500 mg twice daily as one treatment arm. Naproxen was given to 403 patients (280 knee, 123 hip) and placebo to 108 patients (75 knee, 33 hip). WOMAC (Western Ontario and McMaster Universities osteoarthritis index) 3.1 visual analogue scale and SF-36 (36 item short form health survey) were used to assess response to treatment between baseline and week 6. RESULTS: There were no differences at baseline between knee and hip osteoarthritis for any of the WOMAC subscales or SF 36 domains. Improvement was between 4 and 7 mm greater for knee than for hip for all WOMAC subscales (pain, delta = 4.7 mm (p = 0.03); stiffness, delta = 6.6 mm (p = 0.004); function, delta = 4.8 mm (p = 0.06)). Effect size was about 0.8 for all WOMAC subscales for the knee and between 0.5 and 0.6 for the hip. Knee patients treated with naproxen improved 4.6 (p = 0.033) more than hip patients for SF-36 bodily pain and 10.3 (p = 0.014) more for SF-36 role-physical. CONCLUSIONS: Patients with knee osteoarthritis improved more with naproxen treatment than patients with hip osteoarthritis, as monitored by WOMAC and the SF 36 domains bodily pain and role-physical. These findings warrant further investigation and strongly suggest that efficacy of treatment of osteoarthritis of knee and hip should be evaluated separately. PMID- 16269430 TI - The incidences of and consultation rate for lower extremity complaints in general practice. AB - OBJECTIVE: To estimate the incidence and consultation rate of lower extremity complaints in general practice. METHODS: Data were obtained from the Second Dutch National Survey of General Practice, in which 195 general practitioners (GPs) in 104 practices recorded all contacts with patients during 12 consecutive months in computerised patient records. GPs classified the symptoms and diagnosis for each patient at each consultation according to the International Classification of Primary Care (ICPC). Incidence densities and consultation rates for different complaints were calculated. RESULTS: During the registration period 63.2 GP consultations per 1000 person-years were attributable to a new complaint of the lower extremities. Highest incidence densities were seen for knee complaints: 21.4 per 1000 person-years for women and 22.8 per 1000 person-years for men. The incidence of most lower extremity complaints was higher for women than for men and higher in older age. CONCLUSIONS: Both incidences of and consultation rates for lower extremity complaints are substantial in general practice. This implies a considerable impact on the workload of the GP. PMID- 16269429 TI - Systemic lupus erythematosus in a multiethnic US cohort (LUMINA): XXIV. Cytotoxic treatment is an additional risk factor for the development of symptomatic osteonecrosis in lupus patients: results of a nested matched case-control study. AB - BACKGROUND: Osteonecrosis is common in systemic lupus erythematosus (SLE) and often disabling. The role of glucocorticoids in its development is well known. OBJECTIVE: To explore other possible risk factors for osteonecrosis in SLE. METHODS: A nested matched case-control study undertaken in the context of a large, longitudinal, multiethnic lupus cohort (LUMINA), currently formed of 571 SLE patients meeting American College of Rheumatology criteria. All those developing symptomatic osteonecrosis after the diagnosis of SLE were considered cases. Two controls matched for age, disease duration, ethnicity, and centre were selected for each case. Cases and controls were compared by univariable analyses using selected variables. Variables with p<0.10 and those thought clinically relevant were entered into conditional logistic regression models including either the average dose or the highest dose of glucocorticoids, with osteonecrosis as the dependent variable. RESULTS: 32 cases were identified and 59 matched controls selected (in five cases only one control could be found). By univariable analyses, both groups were largely comparable for socioeconomic demographic, clinical, and laboratory variables. Cases were less exposed to hydroxychloroquine (as assessed by the percentage of exposure time) (p = 0.026), used higher doses of glucocorticoids (average and highest doses) (p = 0.011 and 0.001, respectively), and received cytotoxic drugs more often (p = 0.015). In the multivariable analyses only cytotoxic drug use (both models) and the highest dose of glucocorticoids remained associated with the occurrence of osteonecrosis. CONCLUSIONS: Cytotoxic drug use is a risk factor for the development of symptomatic osteonecrosis in SLE patients, along with glucocorticoids. No definite protective factors were identified. PMID- 16269432 TI - Modulation of gene expression and DNA adduct formation in HepG2 cells by polycyclic aromatic hydrocarbons with different carcinogenic potencies. AB - Polycyclic aromatic hydrocarbons (PAHs) can occur in relatively high concentrations in the air, and many PAHs are known or suspected carcinogens. In order to better understand differences in carcinogenic potency between PAHs, we investigated modulation of gene expression in human HepG2 cells after 6 h incubation with varying doses of benzo[a]pyrene (B[a]P), benzo[b]fluoranthene (B[b]F), fluoranthene (FA), dibenzo[a,h]anthracene (DB[a,h]A), 1 methylphenanthrene (1-MPA) or dibenzo[a,l]pyrene (DB[a,l]P), by using cDNA microarrays containing 600 toxicologically relevant genes. Furthermore, DNA adduct levels induced by the compounds were assessed with (32)P-post-labeling, and carcinogenic potency was determined by literature study. All tested PAHs, except 1-MPA, induced gene expression changes in HepG2 cells, although generally no dose-response relationship could be detected. Clustering and principal component analysis showed that gene expression changes were compound specific, since for each compound all concentrations grouped together. Furthermore, it showed that the six PAHs can be divided into three groups, first FA and 1-MPA, second B[a]P, B[b]F and DB[a,h]A, and third DB[a,l]P. This grouping corresponds with the carcinogenic potencies of the individual compounds. Many of the modulated genes are involved in biological pathways like apoptosis, cholesterol biosynthesis and fatty acid synthesis. The order of DNA adduct levels induced by the PAHs was: B[a]P >> DB[a,l]P > B[b]F > DB[a,h]A > 1-MPA >/= FA. When comparing the expression change of individual genes with DNA adduct levels, carcinogenic potency or Ah-receptor antagonicity (the last two were taken from literature), several highly correlated genes were found, of which CYP1A1, PRKCA, SLC22A3, NFKB1A, CYP1A2 and CYP2D6 correlated with all parameters. Our data indicate that discrimination of high and low carcinogenic PAHs by gene expression profiling is feasible. Also, the carcinogenic PAHs induce several pathways that were not affected by the least carcinogenic PAHs. PMID- 16269431 TI - Observational study on efficacy, safety, and drug survival of anakinra in rheumatoid arthritis patients in clinical practice. AB - BACKGROUND: The efficacy and safety of anakinra, a recombinant human interleukin 1 (IL1) receptor antagonist used in rheumatoid arthritis, has been documented in five randomised controlled studies. However, long term post-marketing efficacy data are lacking. OBJECTIVE: To evaluate the efficacy, safety, and drug survival of anakinra in clinical practice. METHODS: All patients with rheumatoid arthritis who started anakinra in six hospitals between May 2002 and February 2004 were included in a two year prospective, in part retrospective, cohort study. Efficacy was assessed using the 28 joint disease activity score (DAS28) and the EULAR response criteria. Safety was evaluated using the common toxicity criteria. Drug survival and prognostic factors were analysed using Kaplan-Meier and Cox proportional hazard analyses. RESULTS: After three months, 55% of the patients (n = 146) showed a response (43% moderate, 12% good). A subset of patients continuing anakinra after 18 months had a sustained clinical response compared with patients who switched to other disease modifying antirheumatic drug treatment (DAS28 improvement, 2.46 v 1.79). Drug survival was 78%, 54%, and 14% after three, six, and 24 months, respectively. The reason for discontinuation was lack of efficacy in 78% and adverse events in 22%. Except for higher drug survival in women (odds ratio = 0.51, 95% confidence interval, 0.27 to 0.97), no prognostic factors were found. Adverse events were reported 206 times in 111 patients, the most common being injection site reactions (36%). Serious adverse events occurred in 12% of the patients, with one classified as related. CONCLUSIONS: The short term efficacy and safety profile of anakinra are comparable to those found in randomised clinical studies. However, the drug survival of anakinra after two years is low, mostly because of lack of efficacy. PMID- 16269433 TI - The influence of mouse Ped gene expression on postnatal development. AB - The Ped (preimplantation embryo development) gene, whose product is Qa-2 protein, is correlated with a faster rate of preimplantation development (Ped fast phenotype) in mice that express Qa-2 protein compared with mice with an absence of Qa-2 protein (Ped slow phenotype). In the current study, we have used two congenic mouse strains differentially expressing the Ped gene, strain B6.K1 (Ped slow; Qa-2 negative) and strain B6.K2 (Ped fast; Qa-2 positive), to investigate the effects of Ped gene expression on postnatal growth profiles, systolic blood pressure and adult organ allometry. At birth, B6.K1 mice were moderately lighter than B6.K2 mice. B6.K1 mice became heavier during postnatal life (P < 0.05) and had elevated systolic blood pressure at 21 weeks of age when compared with B6.K2 mice (P = 0.006). B6.K1 mice also demonstrated elevated serum angiotensin converting enzyme (ACE) activity, a known regulator of blood pressure (P = 0.037). Altered organ:body weight ratios were also observed, with the B6.K1 females having a higher ratio for lungs than B6. K2 females (P = 0.014). These data provide evidence of an association between the rate of preimplantation embryo development, postnatal growth and later cardiovascular function. PMID- 16269435 TI - Excitatory effects of serotonin on rat striatal cholinergic interneurones. AB - We investigated the effects of 5-hydroxytryptamine (5-HT, serotonin) in striatal cholinergic interneurones with gramicidin-perforated whole-cell patch recordings. Bath-application of serotonin (30 microm) significantly and reversibly increased the spontaneous firing rate of 37/45 cholinergic interneurones tested. On average, in the presence of serotonin, firing rate was 273 +/- 193% of control. Selective agonists of 5-HT(1A), 5-HT3, 5-HT4 and 5-HT7 receptors did not affect cholinergic interneurone firing, while the 5-HT2 receptor agonist alpha-methyl-5 HT (30 microm) mimicked the excitatory effects of serotonin. Consistently, the 5 HT2 receptor antagonist ketanserin (10 microm) fully blocked the excitatory effects of serotonin. Two prominent after-hyperpolarizations (AHPs), one of medium duration that was apamin-sensitive and followed individual spikes, and one that was slower and followed trains of spikes, were both strongly and reversibly reduced by serotonin; these effects were fully blocked by ketanserin. Conversely, the depolarizing sags observed during negative current injections and mediated by hyperpolarization-activated cationic currents were not affected. In the presence of apamin and tetrodotoxin, the slow AHP was strongly reduced by 5-HT, and fully abolished by the calcium channel blocker nickel. These results show that 5-HT exerts a powerful excitatory control on cholinergic interneurones via 5-HT2 receptors, by suppressing the AHPs associated with two distinct calcium-activated potassium currents. PMID- 16269434 TI - Transvascular fluid flux from the pulmonary vasculature at rest and during exercise in horses. AB - Exercise causes changes in pulmonary haemodynamics through redistribution of blood flow, increase in the pulmonary surface area, and increase in pulmonary vascular pressures. These changes contribute to the increase in fluid exchange across the alveolar-capillary barrier. To determine the extent of the fluid exchange across the alveolar-capillary barrier at rest and during exercise, six horses were exercised on a high-speed treadmill until fatigue. Arterial and mixed venous blood were sampled at rest and during exercise and recovery. Blood volume changes across the lung (DeltaBV; measured in percentage) were calculated from changes in plasma protein and haemoglobin concentration, and haematocrit. Cardiac output (Q) was calculated using the Fick equation. Fluid flux (J(V-A); measured in l min(-1)) across the alveolar-capillary barrier was then quantified based on Q and DeltaBV. At rest, no fluid movement occurred across the pulmonary vasculature (0.6 +/- 0.6 l min(-1)). During exercise, the amount of fluid moved from the pulmonary circulation was 8.3 +/- 1.3 l min(-1) at 1 min, 6.4 +/- 2.9 l min(-)(1) at 2 min, 10.1 +/- 1.0 l min(-1) at 3 min, 12.9 +/- 2.5 l min(-1) at 4 and 9.6 +/- 1.5 l min(-1) at fatigue (all P < 0.0001). Erythrocyte volume decreased by 6% (P < 0.01) across the lungs, which decreased the colloid osmotic gradient in the pulmonary vasculature. Decrease colloid osmotic gradient along with increased hydrostatic forces in the pulmonary vasculature would enhance displacement of fluid into the pulmonary interstitium. In conclusion, exercise caused large increases in transpulmonary fluid fluxes in horses. Here, we present a simple method to calculate transpulmonary fluid fluxes in different species, which can be used to elucidate mechanisms of lung fluid balance in vivo. PMID- 16269436 TI - Cutaneous reflexes evoked during human walking are reduced when self-induced. AB - Reflex responses are often less pronounced when they are self-induced, but this question has barely been investigated quantitatively. The issue is particularly relevant for locomotion since it has been shown that reflexes elicited during normal gait are important for the regulation of locomotion. The cortex is thought to be involved in the control of reflexes during gait, but it is unclear whether it plays a role in the modulation of these reflexes during the step cycle. During gait, weak electrical stimulation of the sural nerve elicits reflexes in various leg muscles. Are these reflexes different when subjects themselves trigger the stimuli instead of being randomly released by the computer? Cutaneous reflexes were elicited by sural nerve stimulation in 16 phases of the gait cycle in healthy subjects. The stimuli were triggered either by computer or by the subjects themselves. In 6 out of 7 subjects it was observed that the facilitatory responses in leg muscles were smaller and the suppressive responses were more suppressive following self-generated stimuli. In some muscles such as tibialis anterior (TA) both effects were seen (reduced facilitation at end stance and exaggerated suppression at end swing). In all subjects the modulation of anticipatory influences was muscle specific. In the main group of six subjects, the mean reduction in reflex responses was strongest in the TA (max. 30.7%; mean over 16 phases was 12.5%) and weakest in peroneus longus (PL, max. 10.1%; mean over 16 phases was 2.6%). The observation that facilitation is reduced and suppression enhanced in several muscles is taken as evidence that anticipation of self-induced reflex responses reduces the excitatory drive to motoneurones, for example through presynaptic inhibition of facilitatory reflex pathways. PMID- 16269437 TI - Obesity and overweight in relation to disease-specific mortality in men with and without existing coronary heart disease in London: the original Whitehall study. AB - OBJECTIVE: To examine the relations between obesity or overweight and coronary heart disease (CHD) mortality in men with and without prevalent CHD in a prospective cohort study. METHODS: In the Whitehall study of London-based male government employees, 18 403 middle age men were followed up for a maximum of 35 years having participated in a medical examination in the late 1960s in which weight, height, CHD status, and a range of other social, physiological, and behavioural characteristics were measured. RESULTS: In age-adjusted analyses of men with baseline CHD there was a modest raised risk in the overweight relative to normal weight groups for all cause mortality (hazard ratio 1.10, 95% confidence interval (CI) 1.00 to 1.20) and CHD mortality (1.28, 95% CI 1.11 to 1.47) but not for stroke mortality (1.01, 95% CI 0.73 to 1.40). Mortality was similarly raised in the obese group. While these slopes were much steeper in men who were apparently CHD-free at study induction, the difference in the gradients according to baseline CHD status did not attain significance at conventional levels (p value for interaction >or= 0.24). The weight-mortality relations were somewhat attenuated when potential mediating and confounding factors were added to the multivariable models in both men with and men without a history of CHD. CONCLUSIONS: Avoidance of obesity and overweight in adult life in men with and without CHD may reduce their later risk of total and CHD mortality. PMID- 16269438 TI - Right to left shunt through interatrial septal defects in patients with congenital heart disease: results of interventional closure. AB - OBJECTIVE: To study the effects of closure of interatrial communications associated with a right to left shunt in patients with congenital heart disease (CHD) who had a biventricular repair. DESIGN: Retrospective study. SETTING: Tertiary referral centre. PATIENTS: 15 patients with CHD with right to left shunt through an interatrial communication: three had repaired tetralogy of Fallot, five had repaired pulmonary atresia with intact ventricular septum, four had Ebstein's disease, and three had other CHDs. Two patients had had a stroke before closure of the interatrial communication. INTERVENTIONS: Percutaneous atrial septal defect (n = 6) or persistent foramen ovale (n = 9) closure. All patients underwent an exercise test before and after interatrial communication closure. RESULTS: Five patients were cyanotic at rest. During exercise, mean (SD) oxygen saturation diminished from 93.9 (3.8)% to 84.3 (4.8)% (p < 0.05). Interatrial communication closure led to an immediate increase of oxygen saturation from 93.9 (3.8)% to 98.6 (1.6)% (p < 0.05). At a median follow up of three years (range 0.5 5) all but one patient with a residual atrial septal defect had normal oxygen saturation at rest and during exercise. Maximum workload increased from 7.2 (1.9) to 9.0 (2.2) metabolic equivalents (p < 0.001). CONCLUSIONS: Percutaneous closure of interatrial communications associated with a right to left shunt allows restoration of normal oxygen saturation at rest, avoidance of desaturation during exercise, and improvement of exercise performance in patients with CHD. PMID- 16269439 TI - Cerebral autoregulation in neurally mediated syncope: victim or executioner? AB - Involvement of cerebral vasoconstriction confirms the complexity of the pathophysiology of neurally mediated syncope, and the need to adopt a comprehensive approach to the study of this problem. PMID- 16269440 TI - The National Service Framework: six years on. AB - Has the National Service Framework for coronary heart disease resulted in the improvements in cardiac services it promised when first published six years ago? PMID- 16269441 TI - Retinal degeneration associated with RDH12 mutations results from decreased 11 cis retinal synthesis due to disruption of the visual cycle. AB - Retinoid dehydrogenases/reductases catalyze key oxidation-reduction reactions in the visual cycle that converts vitamin A to 11-cis retinal, the chromophore of the rod and cone photoreceptors. It has recently been shown that mutations in RDH12, encoding a retinol dehydrogenase, result in severe and early-onset autosomal recessive retinal dystrophy (arRD). In a cohort of 1011 individuals diagnosed with arRD, we have now identified 20 different disease-associated RDH12 mutations, of which 16 are novel, in a total of 22 individuals (2.2%). Haplotype analysis suggested a founder mutation for each of the three common mutations: p.L99I, p.T155I and c.806_810delCCCTG. Patients typically presented with early disease that affected the function of both rods and cones and progressed to legal blindness in early adulthood. Eleven of the missense variants identified in our study exhibited profound loss of catalytic activity when expressed in transiently transfected COS-7 cells and assayed for ability to convert all-trans retinal to all-trans retinol. Loss-of-function appeared to result from decreased protein stability, as expression levels were significantly reduced. For the p.T49M variant, differing activity profiles were associated with each of the alleles of the common p.R161Q RDH12 polymorphism, suggesting that genetic background may act as a modifier of mutation effect. A locus (LCA3) for Leber congenital amaurosis, a severe, early-onset form of arRD, maps close to RDH12 on chromosome 14q24. Haplotype analysis in the family in which LCA3 was mapped excluded RDH12 as the LCA3 gene and thus suggests the presence of a novel arRD gene in this region. PMID- 16269443 TI - Comprehensive evaluation of common genetic variation within LRRK2 reveals evidence for association with sporadic Parkinson's disease. AB - Parkinson's disease (PD) is a complex neurodegenerative disorder whose aetiologies are largely unknown. To date, mutations in six genes have been found causal for some rare familial forms of the disease and common variation within at least three of these is associated with the more common sporadic forms of PD. LRRK2 is the most recently identified familial PD gene, although its role in sporadic disease is unknown. In this study, we have performed the first comprehensive evaluation of common genetic variation within LRRK2 and investigated its contribution to risk of sporadic PD. We first characterized the linkage disequilibrium within LRRK2 using a panel of densely spaced SNPs across the gene. We then identified a subset of tagging-SNPs (tSNP) that capture the majority of common variation within LRRK2. Both single tSNP and tSNP haplotype analyses, using a large epidemiologically matched sporadic case-control series comprising 932 individuals, yielded significant evidence for disease association. We identified a haplotype that dramatically increases disease risk when present in two copies (OR=5.5, 95%CI=2.1-14.0, P=0.0001). Thus, we provide the first evidence that common genetic variation within LRRK2 contributes to the risk of sporadic PD in the Chinese population. PMID- 16269442 TI - Evaluation of the RET regulatory landscape reveals the biological relevance of a HSCR-implicated enhancer. AB - Evolutionary sequence conservation is now a relatively common approach for the prediction of functional DNA sequences. However, the fraction of conserved non coding sequences with regulatory potential is still unknown. In this study, we focus on elucidating the regulatory landscape of RET, a crucial developmental gene within which we have recently identified a regulatory Hirschsprung disease (HSCR) susceptibility variant. We report a systematic examination of conserved non-coding sequences (n=45) identified in a 220 kb interval encompassing RET. We demonstrate that most of these conserved elements are capable of enhancer or suppressor activity in vitro, and the majority of the elements exert cell type dependent control. We show that discrete sequences within regulatory elements can bind nuclear protein in a cell type-dependent manner that is consistent with their identified in vitro regulatory control. Finally, we focused our attention on the enhancer implicated in HSCR to demonstrate that this element drives reporter expression in cell populations of the excretory system and central nervous system (CNS) and peripheral nervous system (PNS), consistent with expression of the endogenous RET protein. Importantly, this sequence also modulates expression in the enteric nervous system consistent with its proposed role in HSCR. PMID- 16269444 TI - Taking the pulse of health care systems: experiences of patients with health problems in six countries. AB - This paper reports on a 2005 survey of sicker adults in Australia, Canada, Germany, New Zealand, the United Kingdom, and the United States. Sizable shares of patients in all six countries report safety risks, poor care coordination, and deficiencies in care for chronic conditions. Majorities in all countries report that mistakes occurred outside the hospital. The United States often stands out for inefficient care and errors and is an outlier on access/cost barriers. Yet no country consistently leads or lags across survey domains. Deficiencies in transition care during hospital discharge and coordination failures among patients seeing multiple physicians underscore shared challenges of improving performance across sites of care. PMID- 16269445 TI - Blood chimerism in monochorionic twins conceived by induced ovulation: case report. AB - A case of monochorionic twin boys delivered at 34 weeks of gestation following induced ovulation with clomiphene is described. One twin was typed as blood group AB and the other as B. Flow cytometry showed blood group chimerism. DNA polymorphism analysis of peripheral lymphocytes and hair root cells showed that the chimerism was confined to the blood cells and they were dizygotic. PMID- 16269446 TI - Peritoneal fluid leptin is associated with chronic pelvic pain but not infertility in endometriosis patients. AB - BACKGROUND: Leptin influences the proinflammatory immune responses and has angiogenic activity in vitro and in vivo. The objective of this study was to evaluate the peritoneal fluid levels of leptin in patients with endometriosis and idiopathic infertility and compare them with a control group of tubal ligation/reanastomosis patients. METHODS: In this observational, prospective controlled study, peritoneal fluid from 108 women was obtained while they underwent laparoscopy for pelvic pain, infertility, tubal ligation or sterilization reversal. We measured the concentration of leptin in the peritoneal fluid and compared the levels among women who were divided into groups according to their post-surgical diagnosis. Sixty patients were diagnosed with endometriosis, 10 with idiopathic infertility and 38 had undergone tubal ligation or reanastomosis (control group). RESULTS: Peritoneal fluid leptin was significantly higher in endometriosis 14.62+/-9.79 (mean+/-SD) ng/ml compared to idiopathic infertility [0.92+/-1.57 ng/ml (P=0.0007)] and to controls [0.78+/ 1.94 ng/ml (P<0.0001)]. Leptin levels were positively correlated with the stage of endometriosis (r=0.45; P=0.03), and with pelvic pain in endometriosis patients (r=0.49; P=0.001). Peritoneal fluid leptin levels in patients with idiopathic infertility were comparable to controls. CONCLUSIONS: Higher levels of leptin were observed in peritoneal fluid of patients with endometriosis compared to those without the disease. These data suggest that the proinflammatory and neoangiogenic action of leptin may contribute to the pathogenesis of endometriosis. Moreover, leptin may play a role in endometriosis-associated pain. PMID- 16269447 TI - Semen quality of 324 fertile Japanese men. AB - BACKGROUND: A number of studies have indicated regional differences in semen quality. To examine the current status in Japan, we undertook a cross-sectional study on the semen quality of fertile Japanese men for comparison with recent European results. METHODS: Semen parameters of 324 fertile men from the Kawasaki/Yokohama area were investigated. The semen parameters were compared with those published for fertile men from four European cities, Copenhagen, Paris, Edinburgh and Turku. RESULTS: When adjusting for confounders such as ejaculation abstinence period and age, the lowest sperm concentrations were detected in men from Kawasaki/Yokohama followed by men from Copenhagen, Paris, Edinburgh and Turku, but only the differences between men from Kawasaki/Yokohama and men from Edinburgh and Turku were significant (P=0.0008 and P<0.0001, respectively). Total sperm count, percentage of motile sperm and percentage of normal sperm observed in Kawasaki/Yokohama were significantly lower than those from all European centres except for motile sperm in men from Paris. CONCLUSIONS: Japanese fertile men had a semen quality at the level of Danish men, who have been reported to have the lowest among investigated men in Europe. The low level of semen quality of the fertile Japanese men may be due to lifestyle or other environmental factors; however, ethnic differences caused by different genetic variation or combinations cannot be ruled out by this study. PMID- 16269448 TI - Reasons for discontinuation of IVF treatment: a questionnaire study. AB - BACKGROUND: IVF and embryo transfer has become an established and increasingly successful form of treatment for infertility, yet significant numbers of couples discontinue treatment without achieving a live birth. This study aims to identify major factors that influence the decision to discontinue IVF treatment. METHODS: Questionnaires were sent to 1510 couples who had undergone IVF treatment at Ninewells Hospital and Medical School, Dundee, Scotland, between January 1995 and December 2001. The main outcome measure was the number of couples who discontinue treatment and the reasons for discontinuation including live birth, lack of success, lack of funding, psychological stress, medical advice, physical discomfort, personal and other reasons. RESULTS: The response rate was 55% (732/1327) with 183 questionnaires returned as address unknown. A total of 515 couples had discontinued treatment at time of response, with 266 (52%) having achieved a live birth. Achieving a live birth was the reason for discontinuation where a single reason was given. Those who did not conceive gave a combination of reasons. Lack of personal and/or National Health Service funding was cited by 23% of couples as a reason. Lack of success and psychological stress were reported as factors by 23 and 36% of couples respectively. These two factors are very strongly associated (P < 0.001), both being reported by 18% of couples with a reciprocal increase in those quoting lack of success and psychological stress as reasons for discontinuation with increasing number of attempts (P < 0.0005). Changes in personal circumstances were reported by 30% and <10% gave general discomfort or advice from medical staff as reasons. CONCLUSIONS: Though funding is an important issue, factors including lack of success and psychological stress play a greater role in influencing the decision to discontinue treatment. Better information and support are needed to improve the continuation rates. PMID- 16269449 TI - Effect of oral contraceptive pill pretreatment on ongoing pregnancy rates in patients stimulated with GnRH antagonists and recombinant FSH for IVF. A randomized controlled trial. AB - BACKGROUND: The objective of this randomized controlled trial was to assess the effect of oral contraceptive pill (OCP) pretreatment on the probability of ongoing pregnancy in patients treated with a GnRH antagonist for IVF. METHODS: A fixed dose of 200 IU recombinant FSH (rFSH) was started in 425 patients either on day 2 of the menstrual cycle (non-OCP group: n = 211) or 5 days after discontinuing the OCP (OCP group: n = 214). GnRH-antagonist was initiated on day 6 of stimulation, and triggering of final oocyte maturation was performed with 10,000 IU of HCG. RESULTS: Ongoing pregnancy rates per started cycle in the non OCP and OCP group were 27.5% and 22.9%, respectively [95% confidence interval (CI) of the difference: -3.7 to +12.8]. Pregnancy loss was significantly increased in the OCP (36.4%) compared with the non-OCP group (21.6%) (95% CI of the difference: -28.4 to -2.3). CONCLUSION: Pretreatment with OCP, as compared with initiation of stimulation on day 2 of the cycle in patients treated with GnRH antagonist and recombinant FSH, appears to be associated with a not significant difference in ongoing pregnancy rates per started cycle and results in a significantly higher early pregnancy loss. PMID- 16269451 TI - Peroxisome proliferator-activated receptor-gamma transcriptionally up-regulates hormone-sensitive lipase via the involvement of specificity protein-1. AB - Both peroxisome proliferator-activated receptor (PPAR)-gamma and hormone sensitive lipase (HSL) play important roles in lipid metabolism and insulin sensitivity. We demonstrate that expression of the HSL gene is up-regulated by PPARgamma and PPARgamma agonists (rosiglitazone and pioglitazone) in the cultured hepatic cells and differentiating preadipocytes. Rosiglitazone treatment also results in up-regulation of the HSL gene in liver and skeleton muscle from an experimental obese rat model, accompanied by the decreased triglyceride content in these tissues. The proximal promoter (-87 bp of the human HSL gene) was found to be essential for PPARgamma-mediated transactivating activity. This important promoter region contains two GC-boxes and binds the transcription factor specificity protein-1 (Sp1) but not PPARgamma. The Sp1-promoter binding activity can be endogenously enhanced by PPARgamma and rosiglitazone, as demonstrated by analysis of EMSA and chromatin immunoprecipitation assay. Mutations in the GC-box sequences reduce the promoter binding activity of Sp1 and the transactivating activity of PPARgamma. In addition, mithramycin A, the specific inhibitor for Sp1 DNA binding activity, abolishes the PPARgamma-mediated up-regulation of HSL. These results indicate that PPARgamma positively regulates the HSL gene expression, and up-regulation of HSL by PPARgamma requires the involvement of Sp1. Taken together, this study suggests that HSL may be a newly identified PPARgamma target gene, and up-regulation of HSL may be an important mechanism involved in action of PPARgamma agonists in type 2 diabetes. PMID- 16269450 TI - Biological characterization of a heterodimer-selective retinoid X receptor modulator: potential benefits for the treatment of type 2 diabetes. AB - Specific retinoid X receptor (RXR) agonists, such as LG100268 (LG268), and the thiazolidinedione (TZD) PPARgamma agonists, such as rosiglitazone, produce insulin sensitization in rodent models of insulin resistance and type 2 diabetes. In sharp contrast to the TZDs that produce significant increases in body weight gain, RXR agonists reduce body weight gain and food consumption. Unfortunately, RXR agonists also suppress the thyroid hormone axis and generally produce hypertriglyceridemia. Heterodimer-selective RXR modulators have been identified that, in rodents, retain the metabolic benefits of RXR agonists with reduced side effects. These modulators bind specifically to RXR with high affinity and are RXR homodimer partial agonists. Although RXR agonists activate many heterodimer partners, these modulators selectively activate RXR:PPARalpha and RXR:PPARgamma, but not RXR:RARalpha, RXR:LXRalpha, RXR:LXRbeta, or RXR:FXRalpha. We report the in vivo characterization of one RXR modulator, LG101506 (LG1506). In Zucker fatty (fa/fa) rats, LG1506 is a potent insulin sensitizer that also enhances the insulin-sensitizing activities of rosiglitazone. Administration of LG1506 reduces both body weight gain and food consumption and blocks the TZD-induced weight gain when coadministered with rosiglitazone. LG1506 does not significantly suppress the thyroid hormone axis in rats, nor does it elevate triglycerides in Sprague Dawley rats. However, LG1506 produces a unique pattern of triglycerides elevation in Zucker rats. LG1506 elevates high-density lipoprotein cholesterol in humanized apolipoprotein A-1-transgenic mice. Therefore, selective RXR modulators are a promising approach for developing improved therapies for type 2 diabetes, although additional studies are needed to understand the strain-specific effects on triglycerides. PMID- 16269452 TI - Null mutation in transforming growth factor beta1 disrupts ovarian function and causes oocyte incompetence and early embryo arrest. AB - TGFbeta1 is implicated in regulation of ovarian function and the events of early pregnancy. We have investigated the effect of null mutation in the Tgfbeta1 gene on reproductive function in female mice. The reproductive capacity of TGFbeta1 null mutant females was severely impaired, leading to almost complete infertility. Onset of sexual maturity was delayed, after which ovarian function was disrupted, with extended ovarian cycles, irregular ovulation, and a 40% reduction in oocytes ovulated. Serum FSH and estrogen content were normal, but TGFbeta1 null mutant mice failed to display the characteristic proestrus surge in circulating LH. Ovarian hyperstimulation with exogenous gonadotropins elicited normal ovulation rates in TGFbeta1 null mutant mice. After mating with wild-type stud males, serum progesterone content was reduced by 75% associated with altered ovarian expression of mRNAs encoding steroidogenic enzymes 3beta-hydroxysteroid dehydrogenase-1 and P450 17 alpha-hydroxylase/C17-20-lyase. Embryos recovered from TGFbeta1 null mutant females were developmentally arrested in the morula stage and rarely progressed to blastocysts. Attempts to rescue embryos by exogenous progesterone administration and in vitro culture were unsuccessful, and in vitro fertilization and culture experiments demonstrated that impaired development is unlikely to result from lack of maternal tract TGFbeta1. We conclude that embryo arrest is due to developmental incompetence in oocytes developed in a TGFbeta1-deficient follicular environment. This study demonstrates that TGFbeta1 is a critical determinant of normal ovarian function, operating through regulation of LH activity and generation of oocytes competent for embryonic development and successful initiation of pregnancy. PMID- 16269453 TI - Hairless suppresses vitamin D receptor transactivation in human keratinocytes. AB - The vitamin D receptor (VDR) and its ligand 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] are required for normal keratinocyte differentiation. Both the epidermis and the hair follicle are disrupted in VDR-null mice. Hairless (Hr), a presumptive transcription factor with no known ligand, when mutated, disrupts hair follicle cycling similar to the effects of VDR mutations. Hr, like VDR, is found in the nuclei of keratinocytes in both epidermis and hair follicle. To investigate the potential interaction between Hr and VDR on keratinocyte differentiation, we examined the effect of Hr expression on vitamin D-responsive genes in normal human keratinocytes. Inhibition of Hr expression in keratinocytes potentiated the induction of vitamin D-responsive genes, including involucrin, transglutaminase, phospholipase C-gamma1, and 25-hydroxyvitamin D-24-hydroxylase (24-hydroxylase) by 1,25(OH)2D3. Overexpression of Hr in human keratinocytes suppressed the induction of these vitamin D-responsive genes by 1,25(OH)2D3. Coimmunoprecipitation, DNA mobility shift assays, and chromatin immunoprecipitation revealed that Hr binds to VDR in human keratinocytes. Hr binding to the VDR was eliminated by 1,25(OH)2D3, which recruited the coactivator vitamin D receptor-interacting protein 205 (DRIP205) to the VDR/vitamin D response element complex. These data indicate that Hr functions as a corepressor of VDR to block 1,25(OH)2D3 action on keratinocytes. PMID- 16269454 TI - Multiple effects of melatonin on rhythmic clock gene expression in the mammalian pars tuberalis. AB - In mammals, changing day length modulates endocrine rhythms via nocturnal melatonin secretion. Studies of the pituitary pars tuberalis (PT) suggest that melatonin-regulated clock gene expression is critical to this process. Here, we considered whether clock gene rhythms continue in the PT in the absence of melatonin and whether the effects of melatonin on the expression of these genes are temporally gated. Soay sheep acclimated to long photoperiod (LP) were transferred to constant light for 24 h, suppressing endogenous melatonin secretion. Animals were infused with melatonin at 4-h intervals across the final 24 h, and killed 3 h after infusion. The expression of five clock genes (Per1, Per2, Cry1, Rev-erbalpha, and Bmal1) was measured by in situ hybridization. In sham-treated animals, PT expression of Per1, Per2, and Rev-erbalpha showed pronounced temporal variation despite the absence of melatonin, with peak times occurring earlier than predicted under LP. The time of peak Bmal1 expression remained LP-like, whereas Cry1 expression was continually low. Melatonin infusion induced Cry1 expression at all times and suppressed other genes, but only when they showed high expression in sham-treated animals. Hence, 3 h after melatonin treatment, clock gene profiles were driven to a similar state, irrespective of infusion time. In contrast to the PT, melatonin infusions had no clear effect on clock gene expression in the suprachiasmatic nuclei. Our results provide the first example of acute sensitivity of multiple clock genes to one endocrine stimulus and suggest that rising melatonin levels may reset circadian rhythms in the PT, independently of previous phase. PMID- 16269455 TI - Oleylethanolamide activates Ras-Erk pathway and improves myocardial function in doxorubicin-induced heart failure. AB - Oleylethanolamide (OEA) is a natural fatty acid ethanolamide produced in the heart, but its biological actions in myocardium have not yet been defined. This study was carried out to determine whether OEA could be used to prevent the development of heart failure or improve evolving heart failure. We studied in vivo and in vitro actions of OEA in cardiac muscle. In an animal model of doxorubicin cardiomyopathy, OEA showed robust effects and attenuated the progression of systolic/diastolic dysfunction and ventricular remodeling. During evolving doxorubicin cardiomyopathy, a therapeutic course of OEA treatment partially restored myocardial function. The preventive and therapeutic effects of OEA were associated with significant improvement of survival. To investigate the mechanism of OEA action in cardiac muscle, we have carried out in vitro experiments in cultured cardiomyocytes. The results showed that OEA, through activation of Ras-Raf-1-Mek-Erk signaling, inhibited doxorubicin-induced apoptosis. Additional experiments showed that OEA activation of the Erk pathway involved activation of Neu/ErbB2 receptor, which suggests OEA actions in cardiac muscle might require activation of Neu/ErbB2. In summary, OEA improved ventricular remodeling and augmented cardiac function in doxorubicin cardiomyopathy, possibly involving activation of Neu/ErbB2 and Ras-Erk signaling. These findings suggest OEA is a novel cardioprotective compound that may be used to develop new strategies for the management of cardiomyopathy. PMID- 16269457 TI - Intermedin/Adrenomedullin-2 inhibits growth hormone release from cultured, primary anterior pituitary cells. AB - Intermedin (IMD), a novel member of the adrenomedullin (AM), calcitonin gene related peptide (CGRP), amylin (AMY) peptide family, has been reported to act promiscuously at all the known receptors for these peptides. Like AM and CGRP, IMD acts in the circulation to decrease blood pressure and in the brain to inhibit food intake, effects that could be explained by activation of the known CGRP, AM, or AMY receptors. Because AM, CGRP, and AMY have been reported to affect hormone secretion from the anterior pituitary gland, we examined the effects of IMD on GH, ACTH, and prolactin secretion from dispersed anterior pituitary cells harvested from adult male rats. IMD, in log molar concentrations ranging from 1.0 pm to 100 nm, failed to significantly alter basal release of the three hormones. Similarly, IMD failed to significantly alter CRH-stimulated ACTH or TRH-stimulated prolactin secretion in vitro. However, IMD concentration dependently inhibited GHRH-stimulated GH release from these cell cultures. The effects of IMD, although requiring higher concentrations, were as efficacious as those of somatostatin and, like somatostatin, may be mediated, at least in part, by decreasing cAMP accumulation. These actions of IMD were not shared by other members of the AM-CGRP-AMY family of peptides, suggesting the presence of a novel, unique IMD receptor in the anterior pituitary gland and a potential neuroendocrine action of IMD to interact with the hypothalamic mechanisms controlling growth and metabolism. PMID- 16269456 TI - Interleukin-8 synthesis, regulation, and steroidogenic role in H295R human adrenocortical cells. AB - The adrenal gland secretes several cytokines, and cytokines modulate steroid secretion by this gland. In this study, a survey of cytokine production by H295R human adrenocortical cells demonstrated that these cells secreted IL-2, IL-4, IL 8, IL-10, IL-13, and TNFalpha but not IL-5, IL-12, or interferon-gamma. IL-8 was the IL secreted at higher concentration. IL-8 secretion, its regulation, and role in steroidogenesis were further studied. Secreted ILs and steroids were measured by ELISA in cell culture supernatant. IL-8 mRNA was quantified by real-time RT PCR. H295R cells and human adrenal gland expressed IL-8 mRNA. Angiotensin II, potassium, endothelin-1, IL-1alpha, IL-1beta, TNFalpha, and Escherichia coli lipopolysaccharide dose-dependently increase IL-8 secretion by H295R cells after 24 h incubation. IL-6 had no effect on IL-8 secretion. Angiotensin II time dependently increased IL-8 secretion by H295R cells up to 48 h. Angiotensin II caused a biphasic increase in IL-8 mRNA expression with a peak 6 h after stimulation. TNFalpha synergized angiotensin II, potassium, and IL-1alpha mediated IL-8 secretion. IL-8 did not modify aldosterone or cortisol secretion by H295R cells under basal or stimulated (angiotensin II or potassium) conditions. In conclusion, it is demonstrated for the first time that human adrenal cells expressed and secreted IL-8 under the regulation of angiotensin II, potassium, endothelin-1, and immune peptides. Adrenal-secreted IL-8 is one point of convergence between the adrenal gland and the immune system and may have relevance in physiological and pathophysiological conditions associated with increased levels of aldosterone secretagogues and the immune system. PMID- 16269458 TI - Stress-activated signaling pathways mediate the stimulation of pregnancy associated plasma protein-A expression in cultured human fibroblasts. AB - Pregnancy-associated plasma protein-A (PAPP-A) is an IGF binding protein protease that appears to function as a posttranslational modulator of IGF bioavailability in response to injury. A previous study indicated that the proinflammatory cytokines, TNFalpha and IL-1beta, were potent stimulators of PAPP-A expression in cultured human fibroblasts. In this study, we investigated the intracellular signaling pathways mediating cytokine-stimulated PAPP-A expression. Treatment of human fibroblasts with TNFalpha and IL-1beta (1 nm) had little or no effect on phosphatidylinositol 3-kinase and Erk1/2 activation, pathways commonly associated with proliferation. On the other hand, TNFalpha and IL-1beta induced p38, c-Jun N terminal kinase (JNK), and nuclear factor (NF)kappaB activation, pathways more closely related to stress response. An inhibitor of p38 activation (SB203580) had no effect on TNFalpha- or IL-1beta-stimulated PAPP-A expression. The JNK inhibitor, SP600125, had no effect on IL-1beta- or TNFalpha-stimulated PAPP-A mRNA expression. However, SP600125 effectively inhibited IL-1beta-induced PAPP-A protein expression. MG-132, a proteasome inhibitor that blocked degradation of the intrinsic NFkappaB inhibitor, IkappaB, and thereby prevented NFkappaB activation, was a potent inhibitor of both TNFalpha- and IL-1beta-stimulated PAPP A mRNA and protein expression and IGF binding protein-4 protease activity. MG-132 had no effect on JNK phosphorylation or p38 activation, and SB203580 and SP600125 had no effect on IkappaB degradation, documenting inhibitor specificity. BAY11 7082, another inhibitor of NFkappaB activation, also inhibited TNFalpha- and IL 1beta-stimulated PAPP-A expression and IGF binding protein-4 protease activity. These data indicate that NFkappaB activation is the primary mediator of cytokine stimulated PAPP-A expression in human fibroblasts. PMID- 16269459 TI - Altered subcellular distribution of estrogen receptor alpha is implicated in estradiol-induced dual regulation of insulin signaling in 3T3-L1 adipocytes. AB - We investigated the mechanisms by which estrogen alters insulin signaling in 3T3 L1 adipocytes. Treatment with 17beta-estradiol (E2) did not affect insulin induced tyrosine phosphorylation of insulin receptor. E2 enhanced insulin-induced tyrosine phosphorylation of insulin receptor substrate-1 (IRS-1), IRS-1/p85 association, phosphorylation of Akt, and 2-deoxyglucose uptake at 10(-8) m, but inhibited these effects at 10(-5) m. A concentration of 10(-5) m E2 enhanced insulin-induced phosphorylation of IRS-1 at Ser(307), which was abolished by treatment with a c-Jun NH(2)-terminal kinase inhibitor. In addition, the effect of E2 was abrogated by pretreatment with a specific estrogen receptor antagonist, ICI182,780. Membrane-impermeable E2, E2-BSA, did not affect the insulin-induced phosphorylation of Akt at 10(-8) m, but inhibited it at 10(-5) m. Furthermore, E2 decreased the amount of estrogen receptor alpha at the plasma membrane at 10(-8) m, but increased it at 10(-5) m. In contrast, the subcellular distribution of estrogen receptor beta was not altered by the treatment. These results indicate that E2 affects the metabolic action of insulin in a concentration-specific manner, that high concentrations of E2 inhibit insulin signaling by modulating phosphorylation of IRS-1 at Ser(307) via a c-Jun NH(2)-terminal kinase-dependent pathway, and that the subcellular redistribution of estrogen receptor alpha in response to E2 may explain the dual effect of E2. PMID- 16269460 TI - The neuronal growth-associated protein (GAP)-43 is expressed by corticotrophs in the rat anterior pituitary after adrenalectomy. AB - The neuronal growth-associated protein (GAP)-43 has been localized in both long fibers and punctate clusters by immunocytochemistry within the rat anterior pituitary (AP). After adrenalectomy (ADX), GAP-43 immunoreactivity (GAP-43-ir) is greatly increased and is associated with corticotrophs at the light microscopic level. We have undertaken an electron microscopic study to determine the cellular localization of GAP-43 in the post-ADX AP. Using preembedding immunocytochemistry, we found GAP-43-ir localized exclusively to the cytoplasmic surface of the plasmalemma within a subset of endocrine cells with ultrastructure typical of degranulated corticotrophs at 4 d after ADX. We combined preembedding immunoelectron microscopy for GAP-43 with immunogold labeling for ACTH and found that GAP-43-ir was invariably present only in cells containing ACTH-positive granules. The density of GAP-43-ir was highest within extensive processes emanating from the soma, suggesting that these processes are the basis for the punctate clusters of GAP-43 staining seen surrounding corticotrophs in the light microscope. We also observed rare synaptic-like contacts between GAP-43-ir processes and distant cell bodies. GAP-43 mRNA was detected in extracts of the AP 4 d after ADX using RT-PCR, and quantitative PCR confirmed that GAP-43 mRNA was significantly up-regulated in the AP in response to ADX. We postulate that increased expression of GAP-43 may stimulate process outgrowth and intercellular communication by activated corticotrophs. PMID- 16269461 TI - Hyperstimulation and a gonadotropin-releasing hormone agonist modulate ovarian vascular permeability by altering expression of the tight junction protein claudin-5. AB - We investigated the mechanism by which a GnRH agonist (GnRHa) affects ovarian vascularity, vascular permeability, and expression of the tight junction protein claudin-5 in a rat model of ovarian hyperstimulation syndrome (OHSS). Hyperstimulated rats received excessive doses of pregnant mare serum gonadotropin (PMSG; 50 IU/d) for 4 consecutive days, from d 25 to 28 of life, followed by 25 IU human chorionic gonadotropin (hCG) on d 29. Control rats received 10 IU PMSG on d 27 of life, followed by 10 IU hCG on d 29. GnRHa (leuprolide 100 microg/kg.d) was administered to some hyperstimulated rats either on d 29 and 30 (short-term GnRHa treatment) or from d 25 to 30 (long-term GnRHa treatment). Ovarian vascular density (vessels per 10 mm(2)) and vessel endothelial area (percent) were assessed by immunohistochemical analysis of the distribution of von Willebrand factor, whereas vascular permeability was evaluated based on leakage of Evans blue. High doses of PMSG and hCG significantly increased ovarian weight, vascular permeability, vascular density, and the vessel endothelial area and significantly reduced expression of claudin-5 protein and mRNA. All of these effects were significantly and dose-dependently inhibited by administration of GnRHa. This suggests that reduced expression of claudin-5 plays a crucial role in the increased ovarian vascular permeability seen in OHSS and that its expression can be modulated by GnRHa treatment. Indeed, preventing redistribution of tight junction proteins in endothelial cells and the resultant loss of endothelial barrier architecture might be the key to protecting patients against massive extravascular fluid accumulation in cases of OHSS. PMID- 16269462 TI - Thioredoxin-binding protein-2-like inducible membrane protein is a novel vitamin D3 and peroxisome proliferator-activated receptor (PPAR)gamma ligand target protein that regulates PPARgamma signaling. AB - Thioredoxin binding protein-2 (TBP-2), which is identical with vitamin D3 (VD3) up-regulated protein 1 (VDUP1), plays a crucial role in the integration of glucose and lipid metabolism. There are three highly homologous genes of TBP 2/vitamin D3 up-regulated protein 1 in humans, but their functions remain unclear. Here we characterized a TBP-2 homolog, TBP-2-like inducible membrane protein (TLIMP). In contrast to TBP-2, TLIMP displayed no significant binding affinity for thioredoxin. TLIMP exhibited an inner membrane-associated pattern of distribution and also colocalized with transferrin and low-density lipoprotein, indicating endosome- and lysosome-associated functions. VD3 and ligands of peroxisome proliferator-activated receptor (PPAR)-gamma, an important regulator of energy metabolism and cell growth inhibition, induced the expression of TLIMP as well as TBP-2. Overexpression of TLIMP suppressed both anchorage-dependent and -independent cell growth and PPARgamma ligand-inducible gene activation. These results suggest that TLIMP, a novel VD3- or PPARgamma ligand-inducible membrane associated protein, plays a regulatory role in cell proliferation and PPARgamma activation. PMID- 16269463 TI - Disruption of the transmembrane dense core vesicle proteins IA-2 and IA-2beta causes female infertility. AB - Female infertility is a worldwide problem affecting 10-15% of the population. The cause of the infertility in many cases is not known. In the present report, we demonstrate that alterations in two transmembrane structural proteins, IA-2 and IA-2beta, located in dense core secretory vesicles (DCV) of many endocrine and neuroendocrine cells, can result in female infertility. IA-2 and IA-2beta are best known as major autoantigens in type 1 diabetes, but their normal function has remained an enigma. Recently we showed in mice that deletion of IA-2 and/or IA-2beta results in impaired insulin secretion and glucose intolerance. We now report that double knockout (DKO), but not single knockout, female mice are essentially infertile. Vaginal smears showed a totally abnormal estrous cycle, and examination of the ovaries revealed normal-appearing oocytes but the absence of corpora lutea. The LH surge that is required for ovulation occurred in wild type mice but not in DKO mice. Additional studies showed that the LH level in the pituitary of DKO female mice was decreased compared with wild-type mice. Treatment of DKO females with gonadotropins restored corpora lutea formation. In contrast to DKO female mice, DKO male mice were fertile and LH levels in the serum and pituitary were within the normal range. From these studies we conclude that the DCV proteins, IA-2 and IA-2beta, play an important role in LH secretion and that alterations in structural proteins of DCV can result in female infertility. PMID- 16269464 TI - Organ messenger ribonucleic acid and plasma proteome changes in the adjuvant induced arthritis model: responses to disease induction and therapy with the estrogen receptor-beta selective agonist ERB-041. AB - Two receptors [estrogen receptor (ER)alpha and ERbeta] mediate the manifold effects of estrogens throughout the body. Although a clear role has been established for ERalpha in the classical effects of estrogen activity, the physiological role of ERbeta is less well understood. A small-molecule ERbeta selective agonist, ERB-041, has potent antiinflammatory activity in the Lewis rat model of adjuvant-induced arthritis. To characterize the response of target organs and pathways responsible for this antiinflammatory effect, mRNA expression profiling of the spleen, lymph node, and liver was performed, in conjunction with a global analysis of the plasma proteome. We find that the expression of a large number of genes and proteins are altered in the disease model and the majority of these are partially or fully reversed by ERB-041 treatment. Regulated pathways include the acute-phase response, eicosanoid synthesis, fatty acid metabolism, and iron metabolism. In addition, many of the regulated genes and proteins are known to be dysregulated in human rheumatoid arthritis, providing further evidence that the manifestations of the Lewis rat adjuvant-induced arthritis model bear similarity to the human disease. PMID- 16269465 TI - Saturated fatty acids promote endoplasmic reticulum stress and liver injury in rats with hepatic steatosis. AB - Nonalcoholic fatty liver disease is a relatively new hepatic sequela of obesity and type 2 diabetes. The pathogenesis of liver injury and disease progression in nonalcoholic fatty liver disease, however, is poorly understood. The present study examined the hypothesis that the composition of fatty acids in the steatotic liver promotes liver injury. Using dietary models of hepatic steatosis characterized by similar accumulation of total triglyceride but different composition of fatty acids, we show that hepatic steatosis characterized by increased saturated fatty acids is associated with increased liver injury and markers of endoplasmic reticulum stress (e.g. X-box binding protein-1 mRNA splicing and glucose-regulated protein 78 expression). These changes preceded and/or occurred independently of obesity and differences in leptin, TNFalpha, insulin action, and mitochondrial function. In addition, hepatic steatosis characterized by increased saturated fatty acids reduced proliferative capacity in response to partial hepatectomy and increased liver injury in response to lipopolysaccharide. These data suggest that the composition of fatty acids in the steatotic liver is an important determinant of susceptibility to liver injury. PMID- 16269467 TI - Kaiser Permanente's experience of implementing an electronic medical record: a qualitative study. AB - OBJECTIVE: To examine users' attitudes to implementation of an electronic medical record system in Kaiser Permanente Hawaii. DESIGN: Qualitative study based on semistructured interviews. SETTING: Four primary healthcare teams in four clinics, and four specialty departments in one hospital, on Oahu, Hawaii. Shortly before the interviews, Kaiser Permanente stopped implementation of the initial system in favour of a competing one. PARTICIPANTS: Twenty six senior clinicians, managers, and project team members. RESULTS: Seven key findings emerged: users perceived the decision to adopt the electronic medical record system as flawed; software design problems increased resistance; the system reduced doctors' productivity, especially during initial implementation, which fuelled resistance; the system required clarification of clinical roles and responsibilities, which was traumatic for some individuals; a cooperative culture created trade-offs at varying points in the implementation; no single leadership style was optimal--a participatory, consensus-building style may lead to more effective adoption decisions, whereas decisive leadership could help resolve barriers and resistance during implementation; the process fostered a counter climate of conflict, which was resolved by withdrawal of the initial system. CONCLUSIONS: Implementation involved several critical components, including perceptions of the system selection, early testing, adaptation of the system to the larger organisation, and adaptation of the organisation to the new electronic environment. Throughout, organisational factors such as leadership, culture, and professional ideals played complex roles, each facilitating and hindering implementation at various points. A transient climate of conflict was associated with adoption of the system. PMID- 16269466 TI - Insulin receptor kinase-associated phosphotyrosine phosphatases in hepatic endosomes: assessing the role of phosphotyrosine phosphatase-1B. AB - Previous work has shown that bisperoxo(1,10-phenanthroline)-oxovanadate(v) anion [bpV(phen)] induces potent insulin-mimicking effects in the rat, selectively activates the endosomal (EN) insulin receptor kinase (IRK) in liver, and markedly abolishes endosomal IRK-associated phosphotyrosine phosphatase (PTP) activity while reducing that of total ENs by approximately 30%. In this study we examined the relatively selective effect of bpv(phen) on endosomal PTP activities for the purpose of defining IRK-associated PTP(s). Using an in-gel PTP assay, we detected multiple (approximately 20) species of endosomal PTP (30 to >220 kDa), with five that were markedly inhibited after in vivo bpV(phen) administration. Using a combination of Mono Q anionic exchange chromatography and immunoblotting, we demonstrated that LAR (leukocyte common antigen-related), PTP-alpha, and PTP-1B were present in endosomal subfractions not significantly inhibited by bpv(phen). PTP-1B activity was assayed in immunoprecipitates from hepatic ENs of control and bpV(phen)-treated rats and was found to be inhibited by approximately 30% after bpv(phen) treatment. To clarify the role of PTP-1B in dephosphorylating IRK, we prepared hepatic ENs from wild-type and PTP-1B-null mice. We found that the phosphotyrosine content of IRK was similar in these two types of ENs, and that IRK dephosphorylation was not affected in ENs from PTP-1B-null mice compared with that in ENs from wild-type mice. These data suggest that LAR , PTP-alpha, and PTP 1B are not candidates for the IRK-associated PTP in hepatic ENs, and that IRK dephosphorylation in ENs may result from the concerted actions of several PTPs. PMID- 16269469 TI - Patients at risk of familial colorectal cancer. PMID- 16269470 TI - Information for patients on medicines. PMID- 16269471 TI - Adjuvant trastuzumab for breast cancer. PMID- 16269472 TI - Including older people in clinical research. PMID- 16269473 TI - Human resources for health in Africa. PMID- 16269474 TI - UK will review partial smoking ban in 2010, MPs' committee told. PMID- 16269475 TI - UK government is condemned for compromise on smoking ban. PMID- 16269477 TI - Lawyers gain access to Guantanamo medical records. PMID- 16269478 TI - Roche Canada stops distributing oseltamivir. PMID- 16269479 TI - Doctors warn of potentially catastrophic flu pandemic in UK. PMID- 16269480 TI - Experts question wisdom of stockpiling oseltamivir. PMID- 16269487 TI - Romania still faces high abortion rate 16 years after fall of Ceausescu. PMID- 16269489 TI - Distribution anomalies hinder access to flu vaccine in the US. PMID- 16269490 TI - India plans to audit clinical trials. PMID- 16269492 TI - UK must tackle road crashes, skin cancer, and asthma. PMID- 16269493 TI - Snoring. PMID- 16269494 TI - Influenza pandemics and avian flu. PMID- 16269495 TI - Postpartum eclampsia of late onset. PMID- 16269496 TI - Referral or follow-up? PMID- 16269497 TI - Medical paternalism and expensive unsubsidised drugs. PMID- 16269498 TI - Withholding policies from patients restricts their autonomy. PMID- 16269499 TI - Omitted evidence? PMID- 16269500 TI - Details of HPV immunisation need not delay decision. PMID- 16269501 TI - Endovascular repair is not worse than open repair of abdominal aortic aneurysms. PMID- 16269502 TI - Are consenters representative of a target population? PMID- 16269506 TI - Author rectifies omission: health behaviour change in the developing world. PMID- 16269507 TI - Mandatory reporting of all sexually active under-13s: reporting is a public health imperative. PMID- 16269508 TI - Mandatory reporting of all sexually active under-13s: confidential sexual health services to young people: part of the solution or part of the problem? PMID- 16269509 TI - Psychological impact of alopecia: speaking from personal experience. PMID- 16269510 TI - Psychological impact of alopecia: don't forget syphilis. PMID- 16269511 TI - Psychological impact of alopecia: alopecia may lead to social anxiety. PMID- 16269512 TI - Are medical schools fit for graduates? PMID- 16269513 TI - Assay for the measurement of copeptin, a stable peptide derived from the precursor of vasopressin. AB - BACKGROUND: Arginine vasopressin (AVP) is a key regulator of water balance, but its instability makes reliable measurement difficult and precludes routine use. We present a method for quantifying AVP release by use of copeptin, a glycopeptide comprising the C-terminal part of the AVP prohormone. METHODS: We measured copeptin in 50-microL serum and plasma samples from healthy individuals and from critically ill patients with sepsis. Our sandwich immunoluminometric assay used 2 polyclonal antibodies to amino acids 132-164 of pre-provasopressin. RESULTS: The assay yielded results within 3 h. The analytical detection limit was 1.7 pmol/L, and the interlaboratory CV was <20% for values >2.25 pmol/L. The assay was linear on dilution of the analyte. Ex vivo copeptin stability (<20% loss of analyte) for at least 7 days at room temperature and 14 days at 4 degrees C was shown for serum and EDTA-, heparin-, and citrate plasma. Copeptin (median, 4.2 pmol/L; range, 1-13.8 pmol/L) was detectable in 97.5% of 359 healthy individuals and was not associated with age. Median concentrations were considerably higher in men than women, increased significantly after exercise, and were influenced by fasting and water load. Copeptin was significantly (P <0.001) increased in 60 critically ill patients with sepsis (median, 79.5 pmol/L; range, 10.6-228.0 pmol/L). The correlation between copeptin and AVP for 110 samples was r = 0.78 (P <0.0001). CONCLUSIONS: Copeptin is stable for days after blood withdrawal and can be quickly and easily measured. The copeptin assay may be a useful alternative to direct measurement of AVP concentration. PMID- 16269514 TI - Unbound free fatty acids and heart-type fatty acid-binding protein: diagnostic assays and clinical applications. AB - BACKGROUND: A biomarker that reliably detects myocardial ischemia in the absence of necrosis would be useful for initial identification of unstable angina patients and for differentiating patients with chest pain of an etiology other than coronary ischemia, and could provide clinical utility complementary to that of cardiac troponins, the established markers of necrosis. Unbound free fatty acids (FFA(u)) and their intracellular binding protein, heart-type fatty acid binding protein (H-FABP), have been suggested to have clinical utility as indicators of cardiac ischemia and necrosis, respectively. METHODS: We examined results of clinical assessments of FFA(u) and H-FABP as biomarkers of cardiac ischemia and necrosis. Data published on FFA(u) and H-FABP over the past 30 years were used as the basis for this review. RESULTS: Although little clinical work has been done on FFA(u) since the initial reports, recent studies documented an association between increased serum FFAs and ventricular dysrhythmias and death in patients with acute myocardial infarction (AMI). Recent data suggest that serum FFA(u) concentrations increase well before markers of cardiac necrosis and are sensitive indicators of ischemia in AMI. H-FABP is abundant in cardiac muscle and is presumed to be involved in myocardial lipid homeostasis. Similar to myoglobin, plasma H-FABP increases within 3 h after AMI and returns to reference values within 12-24 h. CONCLUSIONS: FFA(u) may have a potential role in identifying patients with cardiac ischemia. H-FABP is useful for detecting cardiac injury in acute coronary syndromes and predicting recurrent cardiac events in acute coronary syndromes and in congestive heart failure patients. Assays are available for both markers that could facilitate further clinical investigations to assess their possible roles as markers of cardiac ischemia and/or necrosis. PMID- 16269517 TI - Mechanism of repression of the inhibin alpha-subunit gene by inducible 3',5' cyclic adenosine monophosphate early repressor. AB - The rodent ovary is regulated throughout the reproductive cycle to maintain normal cyclicity. Ovarian follicular development is controlled by changes in gene expression in response to the gonadotropins FSH and LH. The inhibin alpha-subunit gene belongs to a group of genes that is positively regulated by FSH and negatively regulated by LH. Previous studies established an important role for inducible cAMP early repressor (ICER) in repression of alpha-inhibin. These current studies investigate the mechanisms of repression by ICER. It is not clear whether all four ICER isoforms expressed in the ovary can act as repressors of the inhibin alpha-subunit gene. EMSAs demonstrate binding of all isoforms to the inhibin alpha-subunit CRE (cAMP response element), and transfection studies demonstrate that all isoforms can repress the inhibin alpha-subunit gene. Repression by ICER is dependent on its binding to DNA as demonstrated by mutations to ICER's DNA-binding domain. These mutational studies also demonstrate that repression by ICER is not dependent on heterodimerization with CREB (CRE binding protein). Competitive EMSAs show that ICER effectively competes with CREB for binding to the inhibin alpha CRE in vitro. Chromatin immunoprecipitation assays demonstrate a replacement of CREB dimers bound to the inhibin alpha CRE by ICER dimers in ovarian granulosa cells in response to LH signaling. Thus, there is a temporal association of transcription factors bound to the inhibin alpha-CRE controlling inhibin alpha-subunit gene expression. PMID- 16269515 TI - Two wrongs can make a right: dimers of prolactin and growth hormone receptor antagonists behave as agonists. AB - Prolactin (PRL) and GH have two distinct binding sites (site 1 with high affinity; site 2 with low affinity) that each interact with a PRL receptor (PRLR) to form a functional receptor dimer that activates signal transduction. The G129R mutation in PRL and the G120R mutation in GH disrupt the structural integrity of site 2 such that the ligands retain the ability to bind to the first receptor with high affinity, but act as receptor antagonists. In this study, we examined the ability of monomeric and dimeric forms of these ligands, human (h) PRL and hGH, and their antagonists (hPRL-G129R and hGH-G120R) to 1) bind to PRLRs; 2) induce conformational changes in PRLRs; 3) activate signaling pathways associated with the PRLR; and 4) mediate cell proliferation in vitro. In contrast to monomeric hPRL-G129R, homodimeric hPRL-G129R induced PRLR dimerization; activated Janus family of tyrosine kinases 2/signal transducer and activator of transcription 5, Ras/Raf/MAPK kinase/Erk, and phosphatidylinositol 3-kinase/Akt signaling; and stimulated Nb2 cell proliferation. Similarly, homodimeric hGH G120R was able to mediate signaling via the PRLR and to stimulate Nb2 cell proliferation. These experiments demonstrate that a ligand must have two functional binding sites, but that these may be site 1 plus site 2 or two site 1's, to elicit receptor-mediated signal transduction. The size of the ligand plays less of a role in receptor activation, suggesting that the extracellular portion of the PRLR (and possibly the GH receptor) is rather flexible and can accommodate larger ligands. These findings may have implications for designing multifunctional therapeutics that target this class of cytokine receptors. PMID- 16269516 TI - Follicle-stimulating hormone-induced aromatase in immature rat Sertoli cells requires an active phosphatidylinositol 3-kinase pathway and is inhibited via the mitogen-activated protein kinase signaling pathway. AB - Postnatal development and function of testicular Sertoli cells are regulated primarily by FSH. During this early period of development, estrogens play a role in proliferation of somatic cells, which contributes significantly to testicular development. Growth factors like epidermal growth factor (EGF) are produced in the testis and play a role in regulation of estradiol production and male fertility. Although these divergent factors modulate gonadal function, little is known about their mechanism of action in Sertoli cells. The present study investigates the intracellular events that take place down-stream of FSH and EGF receptors in Sertoli cells isolated from immature (10-d-old) rats, and examines which intracellular signals may be involved in their effects on aromatase activity and estradiol production in immature rat Sertoli cells. Primary cultures of rat Sertoli cells were treated with FSH in combination with EGF and signaling pathway-specific inhibitors. Levels of estradiol production, aromatase mRNA (Cyp19a1), and aromatase protein (CYP19A1) were determined. Western blot analysis was performed to determine the effects of FSH and EGF on levels of activated (phosphorylated) AKT1 and p42 ERK2 and p44 ERK1, also named MAPK1 and MAPK3, respectively. The stimulatory actions of FSH on aromatase mRNA, aromatase protein, and estradiol production were blocked by inhibition of the phosphatidylinositol 3-kinase/AKT1 signaling pathway. In contrast, inhibition of ERK signaling augmented the stimulatory effects of FSH on estradiol production, aromatase mRNA, and protein levels. Furthermore, EGF inhibited the expression of aromatase mRNA and protein in response to FSH, and these inhibitory effects of EGF were critically dependent on the activation of the ERK signaling pathway. We conclude that an active phosphatidylinositol 3-kinase /AKT signaling pathway is required for the stimulatory actions of FSH, whereas an active ERK/MAPK pathway inhibits estradiol production and aromatase expression in immature Sertoli cells. PMID- 16269518 TI - Chronic treatment with prednisolone represses the circadian oscillation of clock gene expression in mouse peripheral tissues. AB - Although altered homeostatic regulation, including disturbance of 24-h rhythms, is often observed in the patients undergoing glucocorticoid therapy, the mechanisms underlying the disturbance remains poorly understood. We report here that chronic treatment with a synthetic glucocorticoid, prednisolone (PSL), can cause alteration of circadian clock function at molecular level. Treatment of cultured hepatic cells (HepG2) with PSL induced expression of Period1 (Per1), and the PSL treatment also attenuated the serum-induced oscillations in the expression of Period2 (Per2), Rev-erbalpha, and Bmal1 mRNA in HepG2 cells. Because the attenuation of clock gene oscillations was blocked by pretreating the cells with a Per1 antisense phosphothioate oligodeoxynucleotide, the extensive expression of Per1 induced by PSL may have resulted in the reduced amplitude of other clock gene oscillations. Continuous administration of PSL into mice constitutively increased the Per1 mRNA levels in liver and skeletal muscle, which seems to attenuate the oscillation in the expressions of Per2, Rev-erbalpha, and Bmal1. However, a single daily administration of PSL at the time of day corresponding to acrophase of endogenous glucocorticoid levels had little effect on the rhythmic expression of clock genes. These results suggest a possible pharmacological action by PSL on the core circadian oscillation mechanism and indicate the possibility that the alteration of clock function induced by PSL can be avoided by optimizing the dosing schedule. PMID- 16269519 TI - The nuclear receptor for bile acids, FXR, transactivates human organic solute transporter-alpha and -beta genes. AB - Bile acids are synthesized from cholesterol in the liver and are excreted into bile via the hepatocyte canalicular bile salt export pump. After their passage into the intestine, bile acids are reabsorbed in the ileum by sodium-dependent uptake across the apical membrane of enterocytes. At the basolateral domain of ileal enterocytes, bile acids are extruded into portal blood by the heterodimeric organic solute transporter OSTalpha/OSTbeta. Although the transport function of OSTalpha/OSTbeta has been characterized, little is known about the regulation of its expression. We show here that human OSTalpha/OSTbeta expression is induced by bile acids through ligand-dependent transactivation of both OST genes by the nuclear bile acid receptor/farnesoid X receptor (FXR). FXR agonists induced endogenous mRNA levels of OSTalpha and OSTbeta in cultured cells, an effect that was not discernible upon inhibition of FXR expression by small interfering RNAs. Furthermore, OST mRNAs were induced in human ileal biopsies exposed to the bile acid chenodeoxycholic acid. Reporter constructs containing OSTalpha or OSTbeta promoters were transactivated by FXR in the presence of its ligand. Two functional FXR binding motifs were identified in the OSTalpha gene and one in the OSTbeta gene. Targeted mutation of these elements led to reduced inducibility of both OST promoters by FXR. In conclusion, the genes encoding the human OSTalpha/OSTbeta complex are induced by bile acids and FXR. By coordinated control of OSTalpha/OSTbeta expression, bile acids may adjust the rate of their own efflux from enterocytes in response to changes in intracellular bile acid levels. PMID- 16269520 TI - Reduction of experimental necrotizing enterocolitis with anti-TNF-alpha. AB - Necrotizing enterocolitis (NEC) is the most common gastrointestinal disease of premature infants. However, despite significant morbidity and mortality, the etiology and pathogenesis of NEC are poorly understood. Evidence suggests that ileal proinflammatory mediators such as IL-18 contribute to the pathology associated with this disease. In addition, we have previously shown that upregulation of TNF-alpha in the liver is correlated with ileal disease severity in a neonatal rat model of NEC. With the use of a neonatal rat model of NEC, we evaluated the incidence and severity of ileal damage along with the production of both hepatic and ileal proinflammatory cytokines in animals injected with (anti TNF-alpha; n = 23) or without (NEC; n = 25) a monoclonal anti-TNF-alpha antibody. In addition, we assessed changes in apoptosis and ileal permeability in the NEC and anti-TNF-alpha groups. Ileal damage was significantly decreased, and the incidence of NEC was reduced from 80% to 17% in animals receiving anti-TNF-alpha. Hepatic TNF-alpha and hepatic and ileal IL-18 were significantly decreased in pups given anti-TNF-alpha compared with those sham injected. In addition, ileal luminal levels of both TNF-alpha and IL-18 were significantly decreased in the anti-TNF-alpha-injected group. Ileal paracellular permeability and the proapoptotic markers Bax and cleaved caspase-3 were significantly decreased in the anti-TNF-alpha group. These data show that hepatic TNF-alpha is an important component for the development of NEC in the neonatal rat model and suggest that anti-TNF-alpha could be used as a potential therapy for human NEC. PMID- 16269521 TI - Defects in cGMP-PKG pathway contribute to impaired NO-dependent responses in hepatic stellate cells upon activation. AB - NO antagonizes hepatic stellate cell (HSC) contraction, although activated HSC in cirrhosis demonstrate impaired responses to NO. Decreased NO responses in activated HSC and mechanisms by which NO affects activated HSC remain incompletely understood. In normal rat HSC, the NO donor diethylamine NONOate (DEAN) significantly increased cGMP production and reduced serum-induced contraction by 25%. The guanylate cyclase (sGC) inhibitor 1H-[1,2,4]oxadiazolo [4,3-a]quinoxalin-1-one (ODQ) abolished 50% of DEAN effects, whereas the cGMP analog 8-bromoguanosine 3',5'-cyclic monophosphate (8-BrcGMP) reiterated half the observed DEAN response, suggesting both cGMP-dependent protein kinase G (PKG) dependent and -independent mechanisms of NO-mediated antagonism of normal HSC contraction. However, NO donors did not increase cGMP production from in vivo activated HSC from bile duct-ligated rats and showed alterations in intracellular Ca(2+) accumulation suggesting defective cGMP-dependent effector pathways. The LX 2 cell line also demonstrated lack of cGMP generation in response to NO and a lack of effect of ODQ and 8-BrcGMP in modulating the NO response. However, cGMP independent effects in response to NO were maintained in LX-2 and were associated with S-nitrosylation of proteins, an effect reiterated in primary HSC. Adenovirus based overexpression of PKG significantly attenuated contraction of LX-2 by 25% in response to 8-BrcGMP. In summary, these studies demonstrate that NO affects HSC through cGMP-dependent and -independent pathways. The HSC activation process is associated with maintenance of cGMP-independent actions of NO but defects in cGMP-PKG-dependent NO signaling that are improved by PKG gene delivery in LX-2 cells. Activating targets downstream from NO-cGMP in activated HSC may represent a novel therapeutic target for portal hypertension. PMID- 16269523 TI - Physical stresses at the air-wall interface of the human nasal cavity during breathing. AB - The nose is the front line defender of the respiratory system and is rich with mechanoreceptors, thermoreceptors, and nerve endings. A time-dependent computational model of transport through nasal models of a healthy human has been used to analyze the fields of physical stresses that may develop at the air-wall interface of the nasal mucosa. Simulations during quiet breathing revealed wall shear stresses as high as 0.3 Pa in the noselike model and 1.5 Pa in the anatomical model. These values are of the same order of those known to exist in uniform large arteries. The distribution of temperature near the nasal wall at peak inspiration is similar to that of wall shear stresses. The lowest temperatures occur in the vicinity of high stresses due to the narrow passageway in these locations. Time and spatial gradients of these stresses may have functional effects on nasal sensation of airflow and may play a role in the well being of nasal breathing. PMID- 16269524 TI - Recovery of phrenic activity and ventilation after cervical spinal hemisection in rats. AB - We tested two hypotheses: 1) that the spontaneous enhancement of phrenic motor output below a C2 spinal hemisection (C2HS) is associated with plasticity in ventrolateral spinal inputs to phrenic motoneurons; and 2) that phrenic motor recovery in anesthetized rats after C2HS correlates with increased capacity to generate inspiratory volume during hypercapnia in unanesthetized rats. At 2 and 4 wk post-C2HS, ipsilateral phrenic nerve activity was recorded in anesthetized, paralyzed, vagotomized, and ventilated rats. Electrical stimulation of the ventrolateral funiculus contralateral to C2HS was used to activate crossed spinal synaptic pathway phrenic motoneurons. Inspiratory phrenic burst amplitudes ipsilateral to C2HS were larger in the 4- vs. 2-wk groups (P<0.05); however, no differences in spinally evoked compound phrenic action potentials could be detected. In unanesthetized rats, inspiratory volume and frequency were quantified using barometric plethysmography at inspired CO2 fractions between 0.0 and 0.07 (inspired O2 fraction 0.21, balance N2) before and 2, 3, and 5 wk post C2HS. Inspiratory volume was diminished, and frequency enhanced, at 0.0 inspired CO2 fraction (P<0.05) 2-wk post-C2HS; further changes were not observed in the 3- and 5-wk groups. Inspiratory frequency during hypercapnia was unaffected by C2HS. Hypercapnic inspiratory volumes were similarly attenuated at all time points post C2HS (P<0.05), thereby decreasing hypercapnic minute ventilation (P<0.05). Thus increases in ipsilateral phrenic activity during 4 wk post-C2HS have little impact on the capacity to generate inspiratory volume in unanesthetized rats. Enhanced crossed phrenic activity post-C2HS may reflect plasticity associated with spinal axons not activated by our ventrolateral spinal stimulation. PMID- 16269522 TI - Modulation of cardiovascular excitatory responses in rats by transcutaneous magnetic stimulation: role of the spinal cord. AB - This study investigated the efficacy of magnetic stimulation on the reflex cardiovascular responses induced by gastric distension in anesthetized rats and compared these responses to those influenced by electroacupuncture (EA). Unilateral magnetic stimulation (30% intensity, 2 Hz) at the Jianshi-Neiguan acupoints (pericardial meridian, P 5-6) overlying the median nerve on the forelimb for 24 min significantly decreased the reflex pressor response by 32%. This effect was noticeable by 20 min of magnetic stimulation and continued for 24 min. Median nerve denervation abolished the inhibitory effect of magnetic stimulation, indicating the importance of somatic afferent input. Unilateral EA (0.3-0.5 mA, 2 Hz) at P 5-6 using similar durations of stimulation similarly inhibited the response (35%). The inhibitory effects of EA occurred earlier and were marginally longer (20 min) than magnetic stimulation. Magnetic stimulation at Guangming-Xuanzhong acupoints (gallbladder meridian, GB 37-39) overlying the superficial peroneal nerve on the hindlimb did not attenuate the reflex. Intravenous naloxone immediately after termination of magnetic stimulation reversed inhibition of the cardiovascular reflex, suggesting involvement of the opioid system. Also, intrathecal injection of delta- and kappa-opioid receptors antagonists, ICI174,864 (n=7) and nor-binaltorphimine (n=6) immediately after termination of magnetic stimulation reversed inhibition of the cardiovascular reflex. In contrast, the mu-opioid antagonist CTOP (n=7) failed to alter the cardiovascular reflex. The endogenous neurotransmitters for delta- and kappa opioid receptors, enkephalins and dynorphin but not beta-endorphin, therefore appear to play significant roles in the spinal cord in mediating magnetic stimulation-induced modulation of cardiovascular reflex responses. PMID- 16269526 TI - Effect of low-repetition jump training on bone mineral density in young women. AB - The hypothesis of the present study was that low-repetition and high-impact training of 10 maximum vertical jumps/day, 3 times/wk would be effective for improving bone mineral density (BMD) in ordinary young women. Thirty-six female college students, with mean age, height, and weight of 20.7+/-0.7 yr, 158.9+/-4.6 cm, and 50.4+/-5.5 kg, respectively, were randomly divided into two groups: jump training and a control group. After the 6 mo of maximum vertical jumping exercise intervention, BMD in the femoral neck region significantly increased in the jump group from the baseline (0.984+/-0.081 vs. 1.010+/-0.080 mg/cm2; P<0.01), although there was no significant change in the control group (0.985+/-0.0143 vs. 0.974+/-0.134 mg/cm2). And also lumbar spine (L2-4) BMD significantly increased in the jump training group from the baseline (0.991+/-0.115 vs. 1.015+/-0.113 mg/cm2; P<0.01), whereas no significant change was observed in the control group (1.007+/-0.113 vs. 1.013+/-0.110 mg/cm2). No significant interactions were observed at other measurement sites, Ward's triangle, greater trochanter, and total hip BMD. Calcium intakes and accelometry-determined physical daily activity showed no significant difference between the two groups. From the results of the present study, low-repetition and high-impact jumps enhanced BMD at the specific bone sites in young women who had almost reached the age of peak bone mass. PMID- 16269525 TI - Predisposition for venoconstriction in the equine laminar dermis: implications in equine laminitis. AB - Equine laminitis is a crippling condition associated with a variety of systemic diseases. Although it is apparent that the prodromal stages of laminitis involve microvascular dysfunction, little is known regarding the physiology of this vasculature. The aim of the present study was to determine the relative responses of equine laminar arteries and veins to the vasoconstrictor agonists phenylephrine (1 nM-10 microM), 5-HT (1 nM-10 microM), PGF2alpha (1 nM-100 microM), and endothelin-1 (1 pM-1 microM). We have determined that laminar veins were more sensitive, with respect to the concentration of agonist required to initiate a contractile response and to achieve EC(50), for all agonists tested. EC50 values, for veins and arteries, respectively, were 84+/-7 vs. 688+/-42 nM for phenylephrine, 35+/-6 vs. 224+/-13 nM for 5-HT, 496+/-43 nM vs. 3.0+/-0.6 microM for PGF2alpha, and 467+/-38 pM vs. 70.6+/-6.4 nM for endothelin-1. Moreover, when expressed as a percentage of the response to a depolarizing stimulus (80 mM potassium), the maximal contractile response of laminar veins exceeded that for the laminar arteries for each agonist. These results indicate that there may be a predisposition for venoconstriction within the vasculature of the equine digit. While this physiological predisposition for venoconstriction may be important in the regulation of blood flow during exercise, it also may help to explain why laminitis can result from a variety of pathological systemic conditions. PMID- 16269527 TI - Aberrant genomic imprinting in rhesus monkey embryonic stem cells. AB - Genomic imprinting involves modification of a gene or a chromosomal region that results in the differential expression of parental alleles. Disruption or inappropriate expression of imprinted genes is associated with several clinically significant syndromes and tumorigenesis in humans. Additionally, abnormal imprinting occurs in mouse embryonic stem cells (ESCs) and in clonally derived animals. Imprinted gene expression patterns in primate ESCs are largely unknown, despite the clinical potential of the latter in the cell-based treatment of human disease. Because of the possible implications of abnormal gene expression to cell or tissue replacement therapies involving ESCs, we examined allele specific expression of four imprinted genes in the rhesus macaque. Genomic and complementary DNA from embryos and ESC lines containing useful single nucleotide polymorphisms were subjected to polymerase chain reaction-based amplification and sequence analysis. In blastocysts, NDN expression was variable indicating abnormal or incomplete imprinting whereas IGF2 and SNRPN were expressed exclusively from the paternal allele and H19 from the maternal allele as expected. In ESCs, both NDN and SNRPN were expressed from the paternal allele while IGF2 and H19 showed loss of imprinting and biallelic expression. In differentiated ESC progeny, these expression patterns were maintained. The implications of aberrant imprinted gene expression to ESC differentiation in vitro and on ESC-derived cell function in vivo after transplantation are unknown. PMID- 16269528 TI - Reduction of Shp-2 expression by small interfering RNA reduces murine embryonic stem cell-derived in vitro hematopoietic differentiation. AB - Shp-2 is a member of a small family of cytoplasmic Src homology 2 (SH2) domain containing protein tyrosine phosphatases. Although Shp-2 has been shown to be necessary for hematopoiesis using a mouse model expressing a mutant residual protein (Shp-2(delta/delta)), we used small interfering RNA (siRNA) to reduce Shp 2 expression and examined the consequences on embryonic stem cell (ESC)-derived hemangioblast, primitive, and definitive hematopoietic development. We found that at a concentration of 50 nM, Shp-2 siRNA effectively diminished Shp-2 expression in differentiating embryoid bodies. Hemangioblast, primitive, and definitive hematopoietic progenitor formation was decreased significantly after transfection with Shp-2 siRNA but not with scrambled siRNA. Because Shp-2 is involved in signals emanating from the basic fibroblast growth factor (bFGF) receptor, we asked whether Shp-2 functions in bFGF-mediated hemangioblast development. Reduction of Shp-2 expression using siRNA, but not scrambled siRNA, blocked the bFGF-induced increase in hemangioblast development. Using siRNA as an independent method of reducing Shp-2 function, in contrast to the mutant mouse model (Shp 2(delta/delta)) previously used, we demonstrate that Shp-2 is required in hemangioblast, primitive, and definitive progenitor hematopoietic development and that Shp-2 is integrally necessary for bFGF-mediated hemangioblast production. PMID- 16269529 TI - Old bone marrow cells inhibit skin wound vascularization. AB - Local injection of hematopoietic stem cell-enriched cells, including mouse lin- cells, accelerates vascularization in animal injury models, apparently by release of angiogenic factors. Locally injected lin- cells from nondiabetic mice dramatically improve, but those from obese diabetic mice inhibit vascular growth in obese diabetic mouse skin wounds. Because of similarities between diabetes and aging and because autologous bone marrow-derived cells are currently being tested in clinical trials involving older patients, we investigated the effects of old lin- cells on skin wound vascularization in nondiabetic and obese diabetic mice. Treatment with old lin- bone marrow cells resulted in decreased vessel size and numerical density, leading to profoundly reduced vascular volume density in wounds of non-diabetic and diabetic mice. Our data suggest that bone marrow derived cells may be poor candidates for therapeutic use in older patients and could actually harm them. PMID- 16269530 TI - Identification of a new class of prostaglandin transporter inhibitors and characterization of their biological effects on prostaglandin E2 transport. AB - Prostaglandins (PGs) are involved in several major signaling pathways. Their effects are terminated when they are transported across cell membranes and oxidized intracellularly. The transport step of PG metabolism is carried out by the prostaglandin transporter (PGT). Inhibition of PGT would therefore be expected to change local or circulating concentrations of prostaglandins, and thus their biological effects. To develop PGT-specific inhibitors with high affinity, we designed a library of triazine compounds and screened 1842 small molecules by using Madin-Darby canine kidney cells stably expressing rat PGT. We found several effective PGT inhibitors. Among them, the most potent inhibitor had a Ki of 3.7 +/- 0.2 microM. These inhibitors allowed us to isolate the efflux process of PGE2 and to demonstrate that PGT does not transport PGE2 outwardly under physiological conditions. PMID- 16269531 TI - Pulmonary bronchiolar cytotoxicity and formation of dichloroacetyl lysine protein adducts in mice treated with trichloroethylene. AB - This study was undertaken to test the hypothesis that bronchiolar damage induced by trichloroethylene (TCE) is associated with bioactivation within the Clara cells with the involvement of CYP2E1 and CYP2F2. Histopathology confirmed dose dependent Clara cell injury and disintegration of the bronchiolar epithelium in CD-1 mice treated with TCE doses of 500 to 1000 mg/kg i.p. Immunohistochemical studies, using an antibody that recognizes dichloroacetyl lysine adducts, revealed dose-dependent formation of adducts in the bronchiolar epithelium. Localization of dichloroacetyl adducts in the Clara cells coincided with damage to this cell type in TCE-treated mice. Pretreatment of CD-1 mice with diallyl sulfone, an inhibitor of CYP2E1 and CYP2F2, abrogated the formation of the dichloroacetyl adducts and protected against TCE-induced bronchiolar cytotoxicity. Treatment of wild-type and CYP2E1-null mice with TCE (750 mg/kg i.p.) also elicited bronchiolar damage that correlated with the formation of adducts in the Clara cells. Immunoblotting, using lung microsomes from TCE treated CD-1 mice, showed dose-dependent production of dichloroacetyl adducts that comigrated with CYP2E1 and CYP2F2. However, TCE treatment resulted in a loss of immunoreactive CYP2E1 and CYP2F2 proteins and p-nitrophenol hydroxylation, a catalytic activity associated with both cytochrome P450 enzymes. The TCE metabolite, chloral hydrate, was formed in incubations of TCE with lung microsomes from CD-1, wild-type, and CYP2E1-null mice. The levels were higher in CD-1 than in either wild-type or CYP2E1-null mice, although levels were higher in CYP2E1-null than in wild-type mice. These findings supported the contention that TCE bioactivation within the Clara cells, predominantly involving CYP2F2, correlated with bronchiolar cytotoxicity in mice. PMID- 16269532 TI - The supply and demand for guns to juveniles: Oakland's gun tracing project. AB - In response to Oakland, California's high level of gun violence affecting young people, the East Oakland Partnership to Reduce Juvenile Gun Violence, a citywide collaboration, was formed in 1997. In 1999, the Partnership established the Oakland Gun Tracing Project to develop evidence-based policy recommendations aimed at reducing the supply of and demand for gun acquisition among urban youth. The advocacy project involved gathering, analyzing, and using police record and gun sale/registration data to inform policy and practice. Such data were collected for all gun crimes committed in Oakland, California between 1998 and 1999 in which a juvenile was either the suspect or the victim. The 213 cases involved 263 juveniles of which 170 were suspects/perpetrators and 93 were victims. Suspects as well as victims were predominantly male and African American. The 213 cases involved 132 recovered guns. Only 55% of the cases were traced to a federally licensed dealer. Three-quarters of the guns were purchased near Oakland, California. Successful traces, defined as the ability to identify federally licensed dealers and initial purchasers, were completed on only 52 of the 132 guns, demonstrating systemic tracing difficulties. Data gathered for the project was used to advocate for numerous policy changes. Recommended policy strategies include initiating a comprehensive gun tracing program so police can track all secondary sales, new laws requiring federal handgun registration which would track ownership changes, required reporting of stolen firearms, and providing effective intervention services to all juveniles the first time they enter the criminal justice system. PMID- 16269533 TI - Volume intake and craving in alcohol withdrawal. AB - AIMS: It has been shown that beer consumption is associated with alcohol craving, in contrast to wine or spirits consumption. The present study was undertaken to evaluate whether the daily volume intake of alcoholic beverages is associated with craving in patients undergoing alcohol withdrawal. METHODS: A total of 158 male patients were assessed using the obsessive compulsive drinking scale (OCDS) at admission. The daily volume intake of alcoholic beverages was calculated by adding the volume of all regularly consumed alcoholic beverages, disregarding their alcohol percentage. Lesch's typology was used to classify patients for subgroup analysis. RESULTS: The daily volume intake of alcoholic beverages correlated significantly with the extent of the OCDS (r = 0.33; P < 0.001). With general linear models, we found a significant association of the calculated daily volume intake of all alcoholic beverages with craving (F = 6.426; P = 0.012), but not for the daily ethanol intake. Differentiating the patients according to Lesch's typology a significant association was particularly found in Lesch Type 2 (model of anxiety) patients (F = 11.31; P = 0.001). CONCLUSION: Our results support the hypothesis that volume intake is associated with craving and suggest a role of pathophysiological changes in volume regulating mechanisms (such as vasopressin or ANP) in the neurobiology of alcohol craving, particularly in male patients of Lesch's Type 2 undergoing alcohol withdrawal. PMID- 16269534 TI - Constraining the expression of nicotinic acetylcholine receptors by using pentameric constructs. AB - Much of our understanding of ligand-gated ion channels comes from heterologous expression studies. However, this technique cannot produce receptors with a predetermined subunit composition for channels formed by several different subunits and cannot insert a single mutation copy if the subunit of interest is present in several copies in the channel. Here, we describe a novel approach that overcomes these problems by expressing pentameric constructs, in which the code of the five subunits is linked (i.e., beta4_beta4_alpha3_beta4_alpha3). This is the first time that a concatemer of the complete pentameric receptor has been expressed for channels in the cysteine-loop superfamily. The presence of the linker did not change the agonist or antagonist sensitivity of alpha3beta4 nicotinic receptors. We show evidence that the expressed receptors were made up of alpha3 and beta4 subunits in one pentameric fusion protein as designed in the construct. This approach can be applied to any nicotinic superfamily receptor to produce channels with a defined subunit arrangement and to introduce specific mutations at any desired location of the pentameric fusion protein. PMID- 16269536 TI - Indirect modulation by alpha7 nicotinic acetylcholine receptors of noradrenaline release in rat hippocampal slices: interaction with glutamate and GABA systems and effect of nicotine withdrawal. AB - Nicotinic acetylcholine receptors (nAChRs) can modulate transmitter release. Striatal [(3)H]dopamine ([(3)H]DA) release is regulated by presynaptic nAChR on dopaminergic terminals and alpha7 nAChR on neighboring glutamatergic afferents. Here, we explored the role of alpha7 nAChR in the modulation of [(3)H]noradrenaline ([(3)H]NA) release from rat hippocampal slices. The nicotinic agonist anatoxin-a (AnTx) evoked monophasic [(3)H]NA release (EC(50) = 1.2 microM) that was unaffected by alpha-conotoxin-MII or dihydro-beta-erythroidine, antagonists of alpha3/alpha6beta2* and beta2* nAChR, respectively. In contrast AnTx-evoked striatal [(3)H]DA release was biphasic (EC(50) = 138.9 nM; 7.1 microM) and blocked by these antagonists. At a high AnTx concentration (25 microM), alpha7 nAChR antagonists (methyllycaconitine, alpha-conotoxin-ImI) and glutamate receptor (GluR) antagonists [kynurenic acid, 6,7-dinitroquinoxaline-2,3 dione (DNQX)] partially inhibited [(3)H]NA release. The alpha7 nAChR-selective agonist choline evoked [(3)H]NA release (E(max) = 33% of that of AnTx) that was blocked by GluR antagonists, supporting a model in which alpha7 nAChRs trigger glutamate release that subsequently stimulates [(3)H]NA release. A GABAergic component was also revealed: choline-evoked [(3)H]NA release was partially blocked by the GABA(A) receptor antagonist bicuculline, and coapplication of bicuculline and DNQX fully abolished this response. These findings support alpha7 nAChR on GABAergic neurons that can promote GABA release which, in turn, leads to [(3)H]NA release, probably by disinhibition. To investigate the impact of long term nicotine exposure on this model, rats were exposed for 14 days to nicotine (4 mg/kg/day) with or without 3 or 7 days of withdrawal. alpha7 nAChR responses were selectively and transiently up-regulated after 3 days of withdrawal. This functional up-regulation could contribute to the withdrawal effects of nicotine. PMID- 16269535 TI - Gene transfer of pro-opiomelanocortin prohormone suppressed the growth and metastasis of melanoma: involvement of alpha-melanocyte-stimulating hormone mediated inhibition of the nuclear factor kappaB/cyclooxygenase-2 pathway. AB - Pro-opiomelanocortin (POMC) is a prohormone of various neuropeptides, including corticotropin, alpha-melanocyte-stimulating hormone (alpha-MSH), and beta endorphin (beta-EP). POMC neuropeptides are potent inflammation inhibitors and immunosuppressants and may exert opposite influences during tumorigenesis. However, the role of POMC expression in carcinogenesis remains elusive. We evaluated the antineoplastic potential of POMC gene delivery in a syngenic B16 F10 melanoma model. Adenovirus-mediated POMC gene delivery in B16-F10 cells increased the release of POMC neuropeptides in cultured media, which differentially regulated the secretion of pro- and anti-inflammatory cytokines in lymphocytes. POMC gene transfer significantly reduced the anchorage-independent growth of melanoma cells. Moreover, pre- or post-treatment with POMC gene delivery effectively retarded the melanoma growth in mice. Intravenous injection of POMC-transduced B16-F10 cells resulted in reduced foci formation in lung by 60 to 70% of control. The reduced metastasis of POMC-transduced B16-F10 cells could be attributed to their attenuated migratory and adhesive capabilities. POMC gene delivery reduced the cyclooxygenase-2 (COX-2) expression and prostaglandin (PG) E(2) synthesis in melanoma cells and tumor tissues. In addition, application of NS-398, a selective COX-2 inhibitor, mimicked the antineoplastic functions of POMC gene transfer in melanoma. The POMC-mediated COX-2 down-regulation was correlated with its inhibition of nuclear factor kappaB (NFkappaB) activities. Exogenous supply of alpha-MSH inhibited NFkappaB activities, whereas application of the alpha-MSH antagonist growth hormone-releasing peptide-6 (GHRP-6) abolished the POMC-induced inhibition of NFkappaB activities and melanoma growth in mice. In summary, POMC gene delivery suppresses melanoma via alpha-MSH-induced inhibition of NFkappaB/COX-2 pathway, thereby constituting a novel therapy for melanoma. PMID- 16269537 TI - Probing the reorganization of the nicotinic acetylcholine receptor during desensitization by time-resolved covalent labeling using [3H]AC5, a photoactivatable agonist. AB - The structural reorganizations occurring on the nicotinic acetylcholine receptor (nAChR) during activation and subsequent desensitization have been investigated through time-resolved photoaffinity labeling using a photoactivatable nicotinic agonist. [(3)H]AC5 is a photosensitive nicotinic probe with high affinity for the desensitized state of the Torpedo marmorata receptor (K(D) = 5 nM) that displays full agonist activity on the Torpedo californica receptor expressed in oocytes (EC(50) = 1.2 microM). Photoaffinity labeling of this receptor in the desensitized state showed a predominant specific labeling of gamma and delta subunits, whereas the alpha subunit was barely labeled. Using a stopped-flow device combined with a flash photolysis quenching system, we investigated the covalent mapping of the subunits as a function of incubation time of the receptor with [(3)H]AC5 (17 ms-1.25 h). During agonist-induced desensitization, specific labeling increased substantially, with similar time constants for gamma and delta subunits (0.016 s(-1)), whereas labeling of the alpha subunit remained relatively low. Therefore, the repartition of radioactivity shifted during desensitization from a weak but predominant labeling of the alpha and gamma subunits toward a substantial labeling of gamma and delta subunits. The observed time-dependent labeling pattern together with AC5 docking into a homology model of the T. californica nAChR suggest a subunit reorganization during agonist-induced desensitization, leading to a tightly packed arrangement that corresponds to a stable high affinity state for agonists. PMID- 16269538 TI - Deleterious mutations in small subunit ribosomal RNA identify functional sites and potential targets for antibiotics. AB - Many clinically useful antibiotics interfere with protein synthesis in bacterial pathogens by inhibiting ribosome function. The sites of action of known drugs are limited in number, are composed primarily of ribosomal RNA (rRNA), and coincide with functionally critical centers of the ribosome. Nucleotide alterations within such sites are often deleterious. To identify functional sites and potential sites of antibiotic action in the ribosome, we prepared a random mutant library of rRNA genes and selected dominant mutations in 16S rRNA that interfere with cell growth. Fifty-three 16S rRNA positions were identified whose mutation inhibits protein synthesis. Mutations were ranked according to the severity of the phenotype, and the detrimental effect of several mutations on translation was verified in a specialized ribosome system. Analysis of the polysome profiles of mutants suggests that the majority of the mutations directly interfered with ribosome function, whereas a smaller fraction of mutations affected assembly of the small ribosomal subunit. Twelve of the identified mutations mapped to sites targeted by known antibiotics, confirming that deleterious mutations can be used to identify antibiotic targets. About half of the mutations coincided with known functional sites in the ribosome, whereas the rest of the mutations affected ribosomal sites with less clear functional significance. Four clusters of deleterious mutations in otherwise unremarkable ribosomal sites were identified, suggesting their functional importance and potential as antibiotic targets. PMID- 16269539 TI - Cooperative water filling of a nonpolar protein cavity observed by high-pressure crystallography and simulation. AB - Formation of a water-expelling nonpolar core is the paradigm of protein folding and stability. Although experiment largely confirms this picture, water buried in "hydrophobic" cavities is required for the function of some proteins. Hydration of the protein core has also been suggested as the mechanism of pressure-induced unfolding. We therefore are led to ask whether even the most nonpolar protein core is truly hydrophobic (i.e., water-repelling). To answer this question we probed the hydration of an approximately 160-A(3), highly hydrophobic cavity created by mutation in T4 lysozyme by using high-pressure crystallography and molecular dynamics simulation. We show that application of modest pressure causes approximately four water molecules to enter the cavity while the protein itself remains essentially unchanged. The highly cooperative filling is primarily due to a small change in bulk water activity, which implies that changing solvent conditions or, equivalently, cavity polarity can dramatically affect interior hydration of proteins and thereby influence both protein activity and folding. PMID- 16269540 TI - An embedded genus-one helicoid. AB - There exists a properly embedded minimal surface of genus one with a single end asymptotic to the end of the helicoid. This genus-one helicoid is constructed as the limit of a continuous one-parameter family of screw-motion invariant minimal surfaces, also asymptotic to the helicoid, that have genus equal to one in the quotient. PMID- 16269541 TI - Parkinson's disease-associated mutations in leucine-rich repeat kinase 2 augment kinase activity. AB - Mutations in the leucine-rich repeat kinase 2 gene (LRRK2) cause late-onset Parkinson's disease (PD) with a clinical appearance indistinguishable from idiopathic PD. Initial studies suggest that LRRK2 mutations are the most common yet identified determinant of PD susceptibility, transmitted in an autosomal dominant mode of inheritance. Herein, we characterize the LRRK2 gene and transcript in human brain and subclone the predominant ORF. Exogenously expressed LRRK2 protein migrates at approximately 280 kDa and is present largely in the cytoplasm but also associates with the mitochondrial outer membrane. Familial linked mutations G2019S or R1441C do not have an obvious effect on protein steady state levels, turnover, or localization. However, in vitro kinase assays using full-length recombinant LRRK2 reveal an increase in activity caused by familial linked mutations in both autophosphorylation and the phosphorylation of a generic substrate. These results suggest a gain-of-function mechanism for LRRK2-linked disease with a central role for kinase activity in the development of PD. PMID- 16269542 TI - A microscopic view of miniprotein folding: enhanced folding efficiency through formation of an intermediate. AB - The role of polypeptide collapse and formation of intermediates in protein folding is still under debate. Miniproteins, small globular peptide structures, serve as ideal model systems to study the basic principles that govern folding. Experimental investigations of folding dynamics of such small systems, however, turn out to be challenging, because requirements for high temporal and spatial resolution have to be met simultaneously. Here, we demonstrate how selective quenching of an extrinsic fluorescent label by the amino acid tryptophan (Trp) can be used to probe folding dynamics of Trp-cage (TC), the smallest protein known to date. Using fluorescence correlation spectroscopy, we monitor folding transitions as well as conformational flexibility in the denatured state of the 20-residue protein under thermodynamic equilibrium conditions with nanosecond time resolution. Besides microsecond folding kinetics, we reveal hierarchical folding of TC, hidden to previous experimental studies. We show that specific collapse of the peptide to a molten globule-like intermediate enhances folding efficiency considerably. A single point mutation destabilizes the intermediate, switching the protein to two-state folding behavior and slowing down the folding process. Our results underscore the importance of preformed structure in the denatured state for folding of even the smallest globular structures. A unique method emerges for monitoring conformational dynamics and ultrafast folding events of polypeptides at the nanometer scale. PMID- 16269543 TI - Age-related changes of nuclear architecture in Caenorhabditis elegans. AB - Mutations in lamins cause premature aging syndromes in humans, including the Hutchinson-Gilford Progeria Syndrome (HGPS) and Atypical Werner Syndrome. It has been shown that HGPS cells in culture undergo age-dependent progressive changes in nuclear architecture. However, it is unknown whether similar changes in nuclear architecture occur during the normal aging process. We have observed that major changes of nuclear architecture accompany Caenorhabditis elegans aging. We found that the nuclear architecture in most nonneuronal cell types undergoes progressive and stochastic age-dependent alterations, such as changes of nuclear shape and loss of peripheral heterochromatin. Furthermore, we show that the rate of these alterations is influenced by the insulin/IGF-1 like signaling pathway and that reducing the level of lamin and lamin-associated LEM domain proteins leads to shortening of lifespan. Our work not only provides evidence for changes of nuclear architecture during the normal aging process of a multicellular organism, but also suggests that HGPS is likely a result of acceleration of the normal aging process. Because the nucleus is vital for many cellular functions, our studies raise the possibility that the nucleus is a prominent focal point for regulating aging. PMID- 16269544 TI - An essential RNase III insertion editing endonuclease in Trypanosoma brucei. AB - RNA editing adds and deletes uridine nucleotides in many preedited mRNAs to create translatable mRNAs in the mitochondria of the parasite Trypanosoma brucei. Kinetoplastid RNA editing protein B3 (KREPB3, formerly TbMP61) is part of the multiprotein complex that catalyzes editing in T. brucei and contains an RNase III motif that suggests nuclease function. Repression of KREPB3 expression, either by RNA interference in procyclic forms (PFs) or by conditional inactivation of an ectopic KREPB3 allele in bloodstream forms (BFs) that lack both endogenous alleles, strongly inhibited growth and in vivo editing in PFs and completely blocked them in BFs. KREPB3 repression inhibited cleavage of insertion editing substrates but not deletion editing substrates in vitro, whereas the terminal uridylyl transferase, U-specific exoribonuclease, and ligase activities of editing were unaffected, and approximately 20S editosomes were retained. Expression of KREPB3 alleles with single amino acid mutations in the RNase III motif had similar consequences. These data indicate that KREPB3 is an RNA editing endonuclease that is specific for insertion sites and is accordingly renamed KREN2 (kinetoplastid RNA editing endonuclease 2). PMID- 16269546 TI - Effect of modulating unfolded state structure on the folding kinetics of the villin headpiece subdomain. AB - Equilibrium Fourier transform infrared (FTIR) and temperature-jump (T-jump) IR spectroscopic techniques were used to study the thermodynamics and kinetics of the unfolding and folding of the villin headpiece helical subdomain (HP36), a small three-helix protein. A double phenylalanine mutant (HP36 F47L, F51L) that destabilizes the hydrophobic core of this protein also was studied. The double mutant is less stable than wild type (WT) and has been shown to contain less residual secondary structure and tertiary contacts in its unfolded state. The relaxation kinetics after a T-jump perturbation were studied for both HP36 and HP36 F47L, F51L. Both proteins exhibited biphasic relaxation kinetics in response to a T-jump. The folding times for the WT (3.23 micros at 60.2 degrees C) and double phenylalanine mutant (3.01 micros at 49.9 degrees C) at the approximate midpoints of their thermal unfolding transitions were found to be similar. The folding time for the WT was determined to be 3.34 mus at 49.9 degrees C, similar to the folding time of the double phenylalanine mutant at that temperature. The double phenylalanine mutant, however, unfolds faster with an unfolding time of 3.01 micros compared with 6.97 micros for the WT at 49.9 degrees C. PMID- 16269545 TI - The structural basis of myotonic dystrophy from the crystal structure of CUG repeats. AB - Myotonic dystrophy (DM) type 1 is associated with an expansion of (>50) CTG repeats within the 3' untranslated region (UTR) of the dystrophin myotonin protein kinase gene (dmpk). In the corresponding mRNA transcript, the CUG repeats form an extended stem-loop structure. The double-stranded RNA of the stem sequesters RNA binding proteins away from their normal cellular targets resulting in aberrant transcription, alternative splicing patterns, or both, thereby leading to DM. To better understand the structural basis of DM type 1, we determined to 1.58-A resolution the x-ray crystal structure of an 18-bp RNA containing six CUG repeats. The CUG repeats form antiparallel double-stranded helices that stack end-on-end in the crystal to form infinite, pseudocontinuous helices similar to the long CUG stem loops formed by the expanded CUG repeats in DM type 1. The CUG helix is very similar in structure to A-form RNA with the exception of the unique U-U mismatches. This structure provides a high-resolution view of a toxic, trinucleotide repeat RNA. PMID- 16269547 TI - Dynamic force spectroscopy of parallel individual Mucin1-antibody bonds. AB - We used atomic force microscopy to measure the binding forces between Mucin1 (MUC1) peptide and a single-chain variable fragment (scFv) antibody selected from a scFv library screened against MUC1. This binding interaction is central to the design of molecules used for targeted delivery of radioimmunotherapeutic agents for prostate and breast cancer treatment. Our experiments separated the specific binding interaction from nonspecific interactions by tethering the antibody and MUC1 molecules to the atomic force microscope tip and sample surface with flexible polymer spacers. Rupture force magnitude and elastic characteristics of the spacers allowed identification of the rupture events corresponding to different numbers of interacting proteins. We used dynamic force spectroscopy to estimate the intermolecular potential widths and equivalent thermodynamic off rates for monovalent, bivalent, and trivalent interactions. Measured interaction potential parameters agree with the results of molecular docking simulation. Our results demonstrate that an increase of the interaction valency leads to a precipitous decline in the dissociation rate. Binding forces measured for monovalent and multivalent interactions match the predictions of a Markovian model for the strength of multiple uncorrelated bonds in a parallel configuration. Our approach is promising for comparison of the specific effects of molecular modifications as well as for determination of the best configuration of antibody-based multivalent targeting agents. PMID- 16269548 TI - Childhood leukaemia in Belarus, Russia, and Ukraine following the Chernobyl power station accident: results from an international collaborative population-based case-control study. AB - BACKGROUND: There is little evidence regarding the risk of leukaemia in children following exposure to radionuclides from the Chernobyl Nuclear Power Plant explosion on April 26, 1986. METHODS: This population-based case-control study investigated whether acute leukaemia is increased among children who were in utero or <6 years of age at the time of the Chernobyl accident. Confirmed cases of leukaemia diagnosed from April 26, 1986 through December 31, 2000 in contaminated regions of Belarus, Russia, and Ukraine were included. Two controls were matched to each case on sex, birth year, and residence. Accumulated absorbed radiation dose to the bone marrow was estimated for each subject. RESULTS: Median estimated radiation doses of participants were <10 mGy. A significant increase in leukaemia risk with increasing radiation dose to the bone marrow was found. This association was most evident in Ukraine, apparent (but not statistically significant) in Belarus, and not found in Russia. CONCLUSION: Taken at face value, these findings suggest that prolonged exposure to very low radiation doses may increase leukaemia risk as much as or even more than acute exposure. However the large and statistically significant dose-response might be accounted for, at least in part, by an overestimate of risk in Ukraine. Therefore, we conclude this study provides no convincing evidence of an increased risk of childhood leukaemia as a result of exposure to Chernobyl radiation, since it is unclear whether the results are due to a true radiation-related excess, a sampling-derived bias in Ukraine, or some combination thereof. However, the lack of significant dose responses in Belarus and Russia also cannot convincingly rule out the possibility of an increase in leukaemia risk at low dose levels. PMID- 16269549 TI - Obesity has a greater impact on cardiovascular mortality in younger men than in older men among non-smoking Koreans. AB - INTRODUCTION: We investigated the effects of age on the relationship between body mass index (BMI) and cardiovascular risk factors and cardiovascular mortality in non-smoking Korean men. METHODS: We performed a prospective cohort study of 246,146 non-smoking Korean men aged 20-69 years at baseline (1992) who were initially without history of cancer or weight change. The associations between BMI and cardiovascular risk factors and mortality during an 9-year follow-up period (2000) were stratified by age group after adjustment for family history, alcohol consumption, exercise habits, and economic status. RESULTS: Calculations of odds ratios (ORs) revealed that younger men (<40 years) with greater BMI (>or=28 kg/m2) were at greater risk of high blood pressure, high blood glucose, and high total cholesterol than older men. The ORs for cardiovascular risk factors associated with greater BMI declined linearly with age. The relative risks for mortality from stroke and from all cardiovascular diseases associated with greater BMI were also higher among younger men and declined linearly with age. CONCLUSIONS: The cardiovascular risk factors and mortality associated with greater BMI were higher among younger than older non-smoking Korean men. These findings indicate that obesity has a greater impact among younger men with respect to premature cardiovascular related deaths. PMID- 16269550 TI - Diagnosis and treatment of prosthetic aortic graft infections: confusion and inconsistency in the absence of evidence or consensus. AB - Prosthetic aortic graft infections represent a major diagnostic and therapeutic challenge. Although a combination of clinical assessment, imaging and microbiological investigations is usually helpful, there are no agreed criteria to confirm a diagnosis. Potential pathogens isolated from superficial specimens may be misleading but influence the choice of antimicrobial agents. Removal of the infected material is strongly recommended. However, this is not always possible in the very debilitated or clinically unstable patient. The choice of which antimicrobial agents to administer as empirical or definitive therapy and the duration of treatment are unclear. A multi-disciplinary group is required to offer guidance, based on what evidence there is, and to provide expert consensus (as is the case for infective endocarditis) to optimize the management of these difficult infections. PMID- 16269552 TI - Impact of tenofovir-containing antiretroviral therapy on chronic hepatitis B in a cohort co-infected with human immunodeficiency virus. AB - OBJECTIVES: We studied the impact of tenofovir disoproxil fumarate, given as an antiretroviral medication, on patients with chronic hepatitis B virus (HBV) co infection. METHODS: The polymerase gene-sequence evolution and quantitative HBV loads (HBVL) were observed for 48 weeks in patients taking tenofovir-containing antiretroviral therapy. The patients were grouped according to baseline strata: high-replicative virus (>6 log copies/mL), low-replicative virus at detectable virus loads (<6 log) and HBs-antigen-positive, HBV-DNA-negative individuals. RESULTS: Thirty-one patients were evaluated. The median decline in 20 patients with high-replicative HBV infection was -5.37 log (range: 3.57-7); 11 out of 20 decreased to undetectable levels (lower limit of detection = < 200 copies/mL) and another three were below 400 copies/mL. Out of six patients with detectable HBV DNA at week 48 (HBVL result: range 3.36-4.32 log(10)), we were able to carry out a re-sequence in four patients. We did not observe relevant emerging resistance mutations, or a relevant virus load re-increase from nadir (>+0.5 log). The patients with low-replicative virus (n = 9) and the baseline DNA-negative patients (n = 2) had an undetectable HBV-DNA at week 48. Two patients became HBeAg-negative; one DNA-negative patient became HBsAg-negative. CONCLUSIONS: Tenofovir is effective in treating HBV infection in HIV patients. Patients with high-replicative virus may benefit from this treatment strategy by a reduction in replicative status, a precondition for improved hepatic function. A few patients showed low-level HBV replication. Indicators for clinical HBV-resistance to tenofovir were not observed. PMID- 16269551 TI - Effect of a beta-lapachone-derived naphthoimidazole on Trypanosoma cruzi: identification of target organelles. AB - OBJECTIVES: Investigation of the mode of action of the naphthoimidazole N1, obtained from the reaction of beta-lapachone with benzaldehyde, which among 45 semi-synthetic derivatives of naphthoquinones isolated from Tabebuia sp. was one of the most active compounds against Trypanosoma cruzi trypomastigotes. METHODS: Quantification of the effect of N1 against the proliferative forms of T. cruzi, and investigation of potential targets in the parasite using electron microscopy and flow cytometry techniques. RESULTS: N1 presented the following order of activity: amastigotes > trypomastigotes > epimastigotes. The effect on intracellular forms was approximately 25 times higher than on macrophages and heart muscle cells. N1-treated parasites presented an abnormal chromatin condensation and mitochondrial damage. In epimastigotes, alterations of reservosomes were observed, and in trypomastigotes, a decrease in the electron density of acidocalcisomes was observed. In epimastigotes, the naphthoimidazole inhibited the activity of succinate cytochrome c reductase. Labelling with rhodamine 123 or Acridine Orange was decreased in both forms treated with N1. CONCLUSIONS: The results suggest that epimastigotes, reservosomes, mitochondrion, and nucleus contain N1 targets. In trypomastigotes, in which reservosomes are absent, the organelles affected by the compound were also the mitochondrion and nucleus, as well as acidocalcisomes, in which the decrease in electron density could be due to the use of polyphosphate as an alternative energy supply. PMID- 16269553 TI - Parental consent for newborn screening in southern Taiwan. AB - OBJECTS: With the advent of genetic technologies, many genetic/metabolic disorders can be detected asymptomatically but might be untreatable, and the benefits and risks of screening for them are not fully known. The purpose of this study is to explore current practice with regard to the parental consent process in newborn screening (NBS). DESIGN: Staff in 23 obstetric clinics/hospitals that conduct NBS in one city of southern Taiwan were interviewed. Using content analysis, 15 interview transcripts, eight completed questionnaires, and other relevant documents from the 23 clinics/hospitals were analysed to reveal the framework of the parental consent process in NBS in southern Taiwan. MAIN MEASURES: Three categories-informed consent, informed dissent, and no informed/consent-were developed to analyse the parental consent process in NBS. RESULTS: The parental consent procedures in NBS and the quality of the information provided before obtaining consent vary widely. Because the traditional NBS was incorporated into routine paediatric practices in most clinics/hospitals, the most frequently encountered consent model is "informed dissent" (60.9%) and "no informed/consent" (30.4%); while an "informed consent" model (45.5%) is the frequent model for screening rare metabolic/genetic disorders. CONCLUSIONS: Specific guidelines to regulate the parental consent process for NBS are essential. Further studies should investigate parental responses to NBS, taking these as the basis on which to establish an informed consent model in Taiwan. PMID- 16269554 TI - Ethical considerations in presymptomatic testing for variant CJD. AB - Variant Creutzfeldt-Jakob disease (vCJD) is a fatal, transmissible, neurodegenerative disorder for which there is currently no effective treatment. vCJD arose from the zoonotic spread of bovine spongiform encephalopathy. There is now compelling evidence for human to human transmission through blood transfusions from presymptomatic carriers and experts are warning that the real epidemic may be yet to come. Imperatives exist for the development of reliable, non-invasive presymptomatic diagnostic tests. Research into such tests is well advanced. In this article the ethical implications of the availability of these tests are elaborated and comparisons drawn with predictive genetic testing for Huntington's disease and screening for HIV. Paramount to considerations is the issue of whom to test, weighing up respect for personal autonomy against obligations to benefit and protect society. A paradigm is proposed similar to that used for HIV screening but with unique features: compulsory testing of all blood/organ donors and individuals undergoing surgery or invasive procedures who have a significant risk of disease transmission. PMID- 16269556 TI - Ethical considerations in the application of preconditioning to solid organ transplantation. AB - The shortage of organs for transplantation has led researchers to look for new techniques to expand the donor pool. Preconditioning strategies have the potential to protect organs from transplant associated injury or may improve the function of substandard organs so that they become suitable for transplantation. Translating this type of technology to the clinical setting raises ethical issues, particularly relating to the deceased donor. It is important that society has the opportunity to discuss the issues raised by implementation of preconditioning strategies before they are implemented rather than as a reaction to them. PMID- 16269558 TI - Human embryonic stem cell research debates: a confucian argument. AB - Human embryonic stem cell research can bring about major biomedical breakthroughs and thus contribute enormously to human welfare, yet it raises serious moral problems because it involves using human embryos for experiment. The "moral status of the human embryo" remains the core of such debates. Three different positions regarding the moral status of the human embryo can be categorised: the "all" position, the "none" position, and the "gradualist" position. The author proposes that the "gradualist" position is more plausible than the other two positions. Confucius's moral principle of jen, which proposes a unique theory of "love of gradation", and the principle of yi, which advocates "due treatment for persons", are then explored. The author then argues that our moral obligations to do good to other living organisms, persons, and our families are different. Putting together the "gradualist" position on the human embryo, and Confucius's theories of "love of gradation" and "due treatment for persons", the author concludes that the early embryo has less ethical significance than the later fetus and adult human. The moral obligation we have toward persons is clearer and stronger than that which we have toward human embryos. Embryo research is justifiable if it brings enormous welfare to human persons that cannot be otherwise achieved. The "love of gradation" requires us, however, to extend love and respect towards other entities according to their different status. We should therefore be very cautious in using human embryos for research, acknowledging the gradualist nature of their moral status. PMID- 16269560 TI - Defining death: when physicians and families differ. PMID- 16269562 TI - The tension between self governance and absolute inner worth in Kant's moral philosophy. AB - The concepts of autonomy as the self governance of individuals and dignity as the inner worth of human beings play an important role in contemporary bioethics. Since both notions are crucial to Immanuel Kant's moral theory, it would be tempting to think that Kantian ethics could ease the friction between the two concepts. It is argued in this paper, however, that this line of thought cannot be supported by Kant's original ideas. While he did make a conscious effort to bring autonomy and dignity together, his emphasis on the absolute inner worth of our collective humanity made it impossible for him to embrace fully the personal self determination of individuals, as it is usually understood in today's liberal thinking. PMID- 16269563 TI - Epistemic paternalism in public health. AB - Receiving information about threats to one's health can contribute to anxiety and depression. In contemporary medical ethics there is considerable consensus that patient autonomy, or the patient's right to know, in most cases outweighs these negative effects of information. Worry about the detrimental effects of information has, however, been voiced in relation to public health more generally. In particular, information about uncertain threats to public health, from-for example, chemicals-are said to entail social costs that have not been given due consideration. This criticism implies a consequentialist argument for withholding such information from the public in their own best interest. In evaluating the argument for this kind of epistemic paternalism, the consequences of making information available must be compared to the consequences of withholding it. Consequences that should be considered include epistemic effects, psychological effects, effects on private decisions, and effects on political decisions. After giving due consideration to the possible uses of uncertain information and rebutting the claims that uncertainties imply small risks and that they are especially prone to entail misunderstandings and anxiety, it is concluded that there is a strong case against withholding of information about uncertain threats to public health. PMID- 16269564 TI - Reproductive cloning combined with genetic modification. AB - Although there is widespread opposition to reproductive cloning, some have argued that its use by infertile couples to have genetically related children would be ethically justifiable. Others have suggested that lesbian or gay couples might wish to use cloning to have genetically related children. Most of the main objections to human reproductive cloning are based on the child's lack of unique nuclear DNA. In the future, it may be possible safely to create children using cloning combined with genetic modifications, so that they have unique nuclear DNA. The genetic modifications could be aimed at giving such children genetic characteristics of both members of the couple concerned. Thus, cloning combined with genetic modification could be appealing to infertile, lesbian, or gay couples who seek genetically related children who have genetic characteristics of both members. In such scenarios, the various objections to human reproductive cloning that are based on the lack of genetic uniqueness would no longer be applicable. The author argues that it would be ethically justifiable for such couples to create children in this manner, assuming these techniques could be used safely. PMID- 16269565 TI - Life extension, human rights, and the rational refinement of repugnance. PMID- 16269566 TI - "Hello, hello--it's English I speak!": a qualitative exploration of patients' understanding of the science of clinical trials. AB - Informed consent may be seriously compromised if patients fail to understand the experimental nature of the trial in which they are participating. Using focus groups, the authors explored how prospective trial participants interpret and understand the science of clinical trials by using patient information sheets relative to their medical condition. An opportunity was provided to hear in the patients' own words how they interpret the information and why there is variable understanding. Respondents struggled to comprehend the meaning and purpose of concepts such as randomisation and double blinding, and found them threatening to their ideas of medical care. Suggestions are made about how to improve the national guidelines on written information for trial participants and pretesting of the information sheets is advocated. PMID- 16269567 TI - How to handle informed consent in longitudinal studies when participants have a limited understanding of the study. AB - Empirical findings from a Swedish longitudinal screening study show that many of the research subjects had a limited understanding of the study. Nevertheless they were satisfied with the understanding they had and found it sufficient for informed continued participation. Were they wrong? In this paper, it is argued that the kind of understanding that is morally required depends partly on the kind of understanding on which the research subjects want to base their decisions, and partly on what kind of knowledge they lack. Researchers must ensure that the information process is not flawed and that participants receive the information they want. To achieve this, new information efforts may be needed. Researchers must also ensure that research subjects have knowledge about aspects of importance to them. Lack of understanding may, however, be the result of conscious choices by research subjects to disregard some of the information because it is not important to them. Such choices should normally be respected. PMID- 16269569 TI - Making sense of dignity. AB - In this review of Leon Kass's Life, liberty and the defense of dignity and Deryck Beyleveld and Roger Brownsword's Human dignity in bioethics and biolaw. I consider the prospects for a theory of dignity as a basis for bioethics research. I argue that dignity theories are worth exploring in more detail, but that research needs to consider both "antitheory" accounts of the language of bioethics, and to give more weight to accounts of dignity as an outcome of holding positive liberties and as something that has a psychological dimension. PMID- 16269568 TI - Keep people informed or leave them alone? A suggested tool for identifying research participants who rightly want only limited information. AB - People taking part in research vary in the extent to which they understand information concerning their participation. Since they may choose to limit the time and effort spent on such information, lack of understanding is not necessarily an ethical problem. Researchers who notice a lack of understanding are in the quandary of not knowing whether this is due to flaws in the information process or to participants' deliberate choices. We argue that the two explanations call for different responses.A tool for identifying those research participants who want limited information is presented. This consists of a restricted number of questions about trust in and appraisal of research, priority of time and privacy, and perception of a duty to participate. It is argued that an important group of participants who purposely lack understanding of the study can be identified with this tool. Some limitations to this approach are also discussed. PMID- 16269570 TI - Differential effect of imipenem treatment on wild-type and NK cell-deficient CD8 knockout mice during acute intra-abdominal injury. AB - CD8 knockout mice depleted of natural killer (NK) cells by treatment with anti asialoGM1 (CD8KO/alphaAsGM1 mice) are resistant to injury caused by cecal ligation and puncture (CLP). However, CLP-induced injury is complex. Potential sources of injury include bacterial dissemination, cecal ischemia, and translocation of bacterial toxins. We treated wild-type and CD8KO/alphaAsGM1 mice with imipenem after CLP to decrease bacterial dissemination. Additional mice were subjected to cecal ligation without puncture of the cecal wall or cecal ligation and removal of cecal contents. Imipenem treatment decreased bacterial counts by at least two orders of magnitude. However, all wild-type mice, whether treated with saline or imipenem, died by 42 h after CLP and exhibited significant hypothermia, metabolic acidosis, and high plasma cytokine concentrations. Wild type mice subjected to cecal ligation without puncture also died, despite very low bacterial counts in blood, but wild-type mice subjected to cecal ligation and washout of cecal contents survived. In CD8KO/alphaAsGM1 mice subjected to CLP, imipenem treatment increased survival from 50% to 100%. After cecal ligation without puncture, long-term survival was 80-90% in CD8KO/alphaAsGM1 mice. Hypothermia, metabolic acidosis, and cytokine production were attenuated in CD8KO/alphaAsGM1 mice compared with wild-type controls. These results indicate that bacterial dissemination is not a major source of injury in wild-type mice after CLP, but the presence of gut flora in the cecal lumen is required for induction of systemic inflammation after cecal injury. CD8KO/alphaAsGM1 mice are resistant to the systemic manifestations of cecal injury. PMID- 16269571 TI - Presynaptic actions of endocannabinoids mediate alpha-MSH-induced inhibition of oxytocin cells. AB - We recently showed that central injections of alpha-melanocyte-stimulating hormone (alpha-MSH) inhibits oxytocin cells and reduces peripheral release of oxytocin, but induces oxytocin release from dendrites. Dendritic oxytocin release can be triggered by agents that mobilize intracellular calcium. Oxytocin, like alpha-MSH, mobilizes intracellular calcium stores in oxytocin cells and triggers presynaptic inhibition of afferent inputs that is mediated by cannabinoids. We hypothesized that this mechanism might underlie the inhibitory effects of alpha MSH. To test this, we recorded extracellularly from identified oxytocin and vasopressin cells in the anesthetized rat supraoptic nucleus (SON). Retrodialysis of a CB1 cannabinoid receptor antagonist to the SON blocked the inhibitory effects of intracerebroventricular injections of alpha-MSH on the spontaneous activity of oxytocin cells. We then monitored synaptically mediated responses of SON cells to stimulation of the organum vasculosum of the lamina terminalis (OVLT); this evoked a mixed response comprising an inhibitory component mediated by GABA and an excitatory component mediated by glutamate, as identified by the effects of bicuculline and 6-cyano-7-nitroquinoxaline-2,3-dione applied to the SON by retrodialysis. Application of CB1 receptor agonists to the SON attenuated the excitatory effects of OVLT stimulation in both oxytocin and vasopressin cells, whereas alpha-MSH attenuated the responses of oxytocin cells only. Thus alpha-MSH can act as a "switch"; it triggers oxytocin release centrally, but at the same time through initiating endocannabinoid production in oxytocin cells inhibits their electrical activity and hence, peripheral secretion. PMID- 16269572 TI - Hindbrain administration of NMDA receptor antagonist AP-5 increases food intake in the rat. AB - Hindbrain administration of MK-801, a noncompetitive N-methyl-D-aspartate (NMDA) channel blocker, increases meal size, suggesting NMDA receptors in this location participate in control of food intake. However, dizocilpine (MK-801) reportedly antagonizes some non-NMDA ion channels. Therefore, to further assess hindbrain NMDA receptor participation in food intake control, we measured deprivation induced intakes of 15% sucrose solution or rat chow after intraperitoneal injection of either saline vehicle or D(-)-2-amino-5-phosphonopentanoic acid (AP5), a competitive NMDA receptor antagonist, to the fourth ventricular, or nucleus of the solitary tract (NTS). Intraperitoneal injection of AP5 (0.05, 0.1, 1.0, 3.0, and 5.0 mg/kg) did not alter 30-min sucrose intake at any dose (10.7 +/ 0.4 ml, saline control) (11.0 +/- 0.8, 11.2 +/- 1.0, 11.2 +/- 1.0, 13.1 +/- 2.2, and 11.0 +/- 1.9 ml, AP5 doses, respectively). Fourth ventricular administration of both 0.2 mug (16.7 +/- 0.6 ml) and 0.4 mug (14.9 +/- 0.5 ml) but not 0.1 and 0.6 mug of AP5 significantly increased 60-min sucrose intake compared with saline (11.2 +/- 0.4 ml). Twenty-four hour chow intake also was increased compared with saline (AP5: 31.5 +/- 0.1 g vs. saline: 27.1 +/- 0.6 g). Furthermore, rats did not increase intake of 0.2% saccharin after fourth ventricular AP5 administration (AP5: 9.8 +/- 0.7 ml, vs. saline: 10.5 +/- 0.5 ml). Finally, NTS AP5 (20 ng/30 nl) significantly increased 30- (AP5: 17.2 +/- 0.7 ml vs. saline: 14.6 +/- 1.7 ml), and 60-min (AP5: 19.4 +/- 0.6 ml vs. saline: 15.5 +/- 1.4 ml) sucrose intake, as well as 24-h chow intake (AP5: 31.6 +/- 0.3 g vs. saline: 26.1 +/- 1.2 g). These results support the hypothesis that hindbrain NMDA receptors participate in control of food intake and suggest that this participation also may contribute to control of body weight over a 24-h period. PMID- 16269573 TI - Excretion of fetal biliverdin by the rat placenta-maternal liver tandem. AB - Fetal liver immaturity is accompanied by active heme catabolism. Thus fetal biliary pigments must be excreted toward the mother by the placenta. To investigate biliverdin handling by the placenta-maternal liver tandem, biliverdin IXalpha was administered to 21-day pregnant rats through the jugular vein or the umbilical artery of an in situ perfused placenta. Jugular administration resulted in the secretion into maternal bile of both bilirubin and biliverdin (3:1). However, when biliverdin was administered to the placenta, most of it was transformed into bilirubin before being transferred to the maternal blood. Injecting Xenopus laevis oocytes with mRNA from rat liver or placenta enhanced their ability to take up biliverdin, which was inhibited by estradiol 17beta-d glucuronide. The expression of three OATP isoforms in this system revealed that they have a varying degrees of ability to transport biliverdin (Oatp1/1a1 > Oatp2/1a4 > Oatp4/1b2). The abundance of their mRNA in rat trophoblast was Oatp1/1a1 >> Oatp4/1b2 > Oatp2/1a4. The expression of biliverdin-IXalpha reductase in rat placenta was detected by RT-PCR/sequencing and Western blot analysis. The relative abundance of biliverdin-IXalpha reductase mRNA (determined by real-time quantitative RT-PCR) was fetal liver > placenta > maternal liver. Common bile duct ligation in the last week of pregnancy induced an upregulation of biliverdin-IXalpha reductase in maternal liver but had no effect on fetal liver and placenta. In conclusion, several members of the OATP family may contribute to the uptake of fetal biliverdin by the rat placenta. Before being transferred to the mother, biliverdin is extensively converted into bilirubin by biliverdin-IXalpha reductase, whose expression is maintained even though bilirubin excretion into maternal bile is impaired. PMID- 16269574 TI - In ovo exposure to monochromatic green light promotes skeletal muscle cell proliferation and affects myofiber growth in posthatch chicks. AB - Our previous studies demonstrated that illumination of chicken embryos with monochromatic green light results in enhanced body and muscle weight at later posthatch stages. In the present study, we investigated the cellular and molecular basis of this phenomenon. First, we showed that on day 6 posthatch, myofibers were more uniform in the in ovo illuminated group than in the control group incubated in the dark, with respect to the number of myofibers displaying diameter values within the range of the mean value. Second, we tested the hypothesis that in ovo illumination causes an increase in the number of myoblasts; this in turn can promote posthatch muscle growth. Indeed, a significant increase in the number of skeletal muscle cells isolated from pectoralis muscle was observed in the in ovo illuminated group on days 1 and 3 posthatch relative to the control group. This increased cell number was accompanied by higher expression levels of Pax7 and myogenin proteins on posthatch days 1 and 3, respectively. A parallel analysis of proliferating cells in the intact muscle further demonstrated a significant increase in the number of cells positive for proliferating cell nuclear antigen in muscle from the in ovo illuminated group. Third, we demonstrated that the transition from fetal- to adult-type myoblasts, normally occurring in late stages of chicken embryogenesis, is initiated earlier in embryos subjected to in ovo green-light illumination. We suggest that the stimulatory effect of in ovo illumination on posthatch muscle growth is the result of enhanced proliferation and differentiation of adult myoblasts and myofiber synchronization. PMID- 16269577 TI - Association of body mass index with suicide mortality: a prospective cohort study of more than one million men. AB - The authors investigated the association of body mass index (BMI) with suicide in a record linkage study based on the Swedish Military Service Conscription Register, the Population and Housing Censuses, and the Cause of Death Register. The cohort studied consisted of 1,299,177 Swedish men who were conscripted in 1968-1999, had their BMI measured at age 18-19 years, and were followed up for as long as 31 years. A strong inverse association was found between BMI and suicide. For each 5-kg/m2 increase in BMI, the risk of suicide decreased by 15% (95% confidence interval: 9, 21). The association was similar when subjects with mental disorder at baseline were excluded from the analysis. BMI-suicide associations were similar in relation to suicide deaths occurring in the first 5 years of follow-up (hazard ratio for each 5-kg/m2 increase in BMI = 0.84, 95% confidence interval: 0.73, 0.96) compared with associations > or = 10 years after baseline (hazard ratio = 0.87, 95% confidence interval: 0.79, 0.96), indicating that weight loss as a consequence of mental illness does not explain the BMI suicide association and that factors influencing BMI may be causally implicated in the etiology of mental disorders leading to suicide. PMID- 16269575 TI - Glucocorticoids act in the dorsal hindbrain to modulate baroreflex control of heart rate. AB - Systemic corticosterone (Cort) modulates arterial baroreflex control of both heart rate and renal sympathetic nerve activity. Because baroreceptor afferents terminate in the dorsal hindbrain (DHB), an area with dense corticosteroid receptor expression, we tested the hypothesis that prolonged activation of DHB Cort receptors increases the midpoint and reduces the gain of arterial baroreflex control of heart rate in conscious rats. Small (3-4 mg) pellets of Cort (DHB Cort) or Silastic (DHB Sham) were placed on the surface of the DHB, or Cort was administered systemically by placing a Cort pellet on the surface of the dura (Dura Cort). Baroreflex control of heart rate was determined in conscious male Sprague Dawley rats on each of 4 days after initiation of treatment. Plots of arterial pressure vs. heart rate were analyzed using a four-parameter logistic function. After 3 days of treatment, the arterial pressure midpoint for baroreflex control of heart rate was increased in DHB Cort rats (123 +/- 2 mmHg) relative to both DHB Sham (108 +/- 3 mmHg) and Dura Cort rats (109 +/- 2 mmHg, P < 0.05). On day 4, baseline arterial pressure was greater in DHB Cort (112 +/- 2 mmHg) compared with DHB Sham (105 +/- 2 mmHg) and Dura Cort animals (106 +/- 2 mmHg, P < 0.05), and the arterial pressure midpoint was significantly greater than mean arterial pressure in the DHB Cort group only. Also on day 4, maximum baroreflex gain was reduced in DHB Cort (2.72 +/- 0.12 beats x min(-1) x mmHg( 1)) relative to DHB Sham and Dura Cort rats (3.51 +/- 0.28 and 3.37 +/- 0.27 beats x min(-1) x mmHg(-1), P < 0.05). We conclude that Cort acts in the DHB to increase the midpoint and reduce the gain of the heart rate baroreflex function. PMID- 16269579 TI - Prospective study of occupation and amyotrophic lateral sclerosis mortality. AB - Occupational exposures are suspected of contributing to the risk of amyotrophic lateral sclerosis (ALS), but results of epidemiologic studies have been inconsistent. The authors prospectively assessed the relation between occupation and ALS mortality among more than 1 million participants in the Cancer Prevention Study II of the American Cancer Society. Follow-up from 1989 through 2002 identified 507 ALS deaths among men and 430 among women. Adjusted rate ratios were calculated by using Mantel-Haenszel weights and Cox proportional hazards. Among men, elevated ALS mortality was found for programmers (rate ratio = 4.55, 95% confidence interval: 1.46, 14.2; p = 0.009) and laboratory technicians (rate ratio = 1.96, 95% confidence interval: 1.04, 3.66; p = 0.04). Occupations previously associated with increased risk of ALS for which no increased risk was found included farmers, electricians, and welders, although the numbers of electricians (eight ALS deaths) and welders (two ALS deaths) were small. Among women, only machine assemblers had significantly increased ALS mortality (rate ratio = 2.81, 95% confidence interval: 1.05, 7.53; p = 0.04). Results, which suggest that male programmers and laboratory technicians and female machine assemblers may be at increased risk of death from ALS, should be interpreted cautiously, however, because they are based on small numbers. PMID- 16269576 TI - NO-dependent blood pressure regulation in RGS2-deficient mice. AB - The regulator of G protein signaling (RGS) 2, a GTPase-activating protein, is activated via the nitric oxide (NO)-cGMP pathway and thereby may influence blood pressure regulation. To test that notion, we measured mean arterial blood pressure (MAP) and heart rate (HR) with telemetry in N(omega)-nitro-l-arginine methyl ester (l-NAME, 5 mg l-NAME/10 ml tap water)-treated RGS2-deficient (RGS2( /-)) and RGS2-sufficient (RGS2(+/+)) mice and assessed autonomic function. Without l-NAME, RGS2(-/-) mice showed during day and night a similar increase of MAP compared with controls. l-NAME treatment increased MAP in both strains. nNOS is involved in this l-NAME-dependent blood pressure increase, since 7 nitroindazole increased MAP by 8 and 9 mmHg (P < 0.05) in both strains. The l NAME-induced MAP increase of 14-15 mmHg during night was similar in both strains. However, the l-NAME-induced MAP increase during the day was smaller in RGS2(-/-) than in RGS2(+/+) (11 +/- 1 vs. 17 +/- 2 mmHg; P < 0.05). Urinary norepinephrine and epinephrine excretion was higher in RGS2(-/-) than in RGS2(+/+) mice. The MAP decrease after prazosin was more pronounced in l-NAME-RGS2(-/-). HR variability parameters [root mean square of successive differences (RMSSD), low-frequency (LF) power, and high-frequency (HF) power] and baroreflex sensitivity were increased in RGS2(-/-). Atropine and atropine plus metoprolol markedly reduced RMSSD, LF, and HF. Our data suggest an interaction between RGS2 and the NO-cGMP pathway. The blunted l-NAME response in RGS2(-/-) during the day suggests impaired NO signaling. The MAP increases during the active phase in RGS2(-/-) mice may be related to central sympathetic activation and increased vascular adrenergic responsiveness. PMID- 16269580 TI - Physical activity, obesity, energy intake, and the risk of non-Hodgkin's lymphoma: a population-based case-control study. AB - The authors conducted a population-based case-control study of 1,030 cases with histologically confirmed, incident non-Hodgkin's lymphoma (NHL) and 3,106 controls to assess the impact of recreational physical activity, obesity, and energy intake on NHL risk in Canada from 1994 to 1997. Compared with those for subjects in the lowest quartiles of total recreational physical activity, multivariable-adjusted odds ratios for subjects in the highest quartile were 0.79 (95% confidence interval (CI): 0.59, 1.05) for men and 0.59 (95% CI: 0.42, 0.81) for women. Obesity (body mass index > or = 30 kg/m2) was associated with odds ratios of 1.59 (95% CI: 1.18, 2.12) for men and 1.36 (95% CI: 1.00, 1.84) for women. For men and women with a lifetime maximum body mass index of > or = 30 kg/m2, respective odds ratios were 1.55 (95% CI: 1.16, 2.06) and 1.10 (95% CI: 0.83, 1.46). For men and women in the highest quartiles of calorie intake, respective odds ratios were 1.95 (95% CI: 1.45, 2.62) and 1.13 (95% CI: 0.84, 1.52). Some differences were found between histologic subtypes of NHL for these associations. This study suggests that recreational physical activity decreases NHL risk, while obesity and excess calorie intake increase it. More studies are needed to confirm these results, especially the differences between histologic subtypes. PMID- 16269581 TI - Two-stage case-control studies: precision of parameter estimates and considerations in selecting sample size. AB - A two-stage case-control design, in which exposure and outcome are determined for a large sample but covariates are measured on only a subsample, may be much less expensive than a one-stage design of comparable power. However, the methods available to plan the sizes of the stage 1 and stage 2 samples, or to project the precision/power provided by a given configuration, are limited to the case of a binary exposure and a single binary confounder. The authors propose a rearrangement of the components in the variance of the estimator of the log-odds ratio. This formulation makes it possible to plan sample sizes/precision by including variance inflation factors to deal with several confounding factors. A practical variance bound is derived for two-stage case-control studies, where confounding variables are binary, while an empirical investigation is used to anticipate the additional sample size requirements when these variables are quantitative. Two methods are suggested for sample size planning based on a quantitative, rather than binary, exposure. PMID- 16269582 TI - Oxidative stress and pulmonary function in the general population. AB - Studies have shown increased oxidative stress in patients with chronic airflow limitation; however, the population-based evidence for the association of oxidative stress with pulmonary function is limited. The authors analyzed the association of plasma thiobarbituric acid-reactive substances (TBARS), glutathione, glutathione peroxidase, and 6-hydroxy-2,5,7,8-tetramethylchroman-2 carboxylic acid (Trolox)-equivalent antioxidant capacity with forced expiratory volume in 1 second and forced vital capacity using data collected from 1996 to 2000 in a general population sample from western New York State (n = 2,346). After adjustment for covariates including smoking status, lifetime pack-years of smoking, education, weight, and eosinophils, multivariate analysis showed an inverse association of TBARS with forced expiratory volume in 1 second and forced vital capacity as the percentage of the predicted value (FEV1% and FVC%, respectively), positive associations of glutathione peroxidase with FEV1% and FVC%, and an inverse association of glutathione with FEV1% in men (p < 0.05). The associations of TBARS and glutathione peroxidase with FVC% in men remained statistically significant after adjustment for serum carotenoid levels. There were no statistically significant associations of oxidative stress with pulmonary function in women. These results suggest that oxidative stress may be associated with airflow limitation in men, and that gender differences may exist in the relation of oxidative stress to pulmonary function. PMID- 16269583 TI - Endothelial nitric oxide synthase (NOS3) genetic variants, maternal smoking, vitamin use, and risk of human orofacial clefts. AB - Orofacial clefts have been associated with maternal cigarette smoking and lack of folic acid supplementation (which results in higher plasma homocysteine concentrations). Because endothelial nitric oxide synthase (NOS3) activity influences homocysteine concentration and because smoking compromises NOS3 activity, genetic variation in NOS3 might interact with smoking and folic acid use in clefting risk. The authors genotyped 244 infants with isolated cleft lip with or without cleft palate (CL/P), 99 with isolated cleft palate, and 588 controls from a California population-based case-control study (1987-1989 birth cohort) for two NOS3 polymorphisms: A(-922)G and G894T. Analyses of gene-only effects for each polymorphism revealed a 60% increased risk of CL/P among NOS3 A( 922)G homozygotes (odds ratio (OR) = 1.6, 95% confidence interval (CI): 1.0, 2.6). There was some evidence for higher risk of CL/P with maternal periconceptional smoking in infants with an NOS3 -922G allele (for homozygotes, OR = 2.5, 95% CI: 1.2, 5.6) but not in those with an 894T allele. For CL/P risk, odds ratios were over 4 among mothers who smoked, who did not use vitamins, and whose infants had at least one variant allele for each NOS3 polymorphism (for A( 922)G, OR = 4.6, 95% CI: 2.1, 10.2; for 894T, OR = 4.4, 95% CI: 1.8, 10.7). No similar patterns were observed for risk of cleft palate. PMID- 16269584 TI - Inflammation and triglycerides partially mediate the effect of prepregnancy body mass index on the risk of preeclampsia. AB - The objective of this study was to quantify the mediating role of inflammation and triglycerides in the association between prepregnancy body mass index (weight (kg)/height (m)2) and preeclampsia. The authors conducted a nested case-control study of 55 preeclamptic women and 165 pregnant controls from the Pregnancy Exposures and Preeclampsia Prevention Study (Pittsburgh, Pennsylvania, 1997 2001). Serum samples collected at < or = 20 weeks' gestation were analyzed for levels of C-reactive protein and triglycerides. The adjusted odds ratio (AOR) from a multivariable conditional logistic regression model assessing the total effect of body mass index on preeclampsia risk was compared with the AOR from the same model after results were controlled for C-reactive protein, triglycerides, and confounding factors (direct-effects model). The percentage of the total effect that was mediated through inflammation and triglycerides was calculated as 100 - [ln(direct-effects AOR)/ln(total-effects AOR)]. In the total-effects model, 4- and 8-unit increases in body mass index were associated with 1.7-fold (95% confidence interval (CI): 1.3, 2.3) and 2.9-fold (95% CI: 1.6, 5.2) increases in preeclampsia risk, whereas in the direct-effects model, these AORs were 1.4 (95% CI: 1.0, 1.9) and 2.0 (95% CI: 1.0, 3.8), respectively. Inflammation was a more important mediator than triglycerides. These findings suggest that approximately one third of the total effect of body mass index on preeclampsia risk is mediated through inflammation and triglyceride levels. PMID- 16269585 TI - Arsenic exposure and cardiovascular disease: a systematic review of the epidemiologic evidence. AB - Arsenic exposure is a likely cause of blackfoot disease and a potential risk factor for atherosclerosis. The authors performed a systematic review of the epidemiologic evidence on the association between arsenic and cardiovascular outcomes. The search period was January 1966 through April 2005. Thirteen studies conducted in general populations (eight in high-arsenic areas in Taiwan, five in other countries) and 16 studies conducted in occupational populations were identified. Exposure was assessed ecologically in most studies. In Taiwan, relative risks comparing the highest arsenic exposure category with the lowest ranged from 1.59 to 4.90 for coronary disease, from 1.19 to 2.69 for stroke, and from 1.66 to 4.28 for peripheral arterial disease. In other general populations, relative risks ranged from 0.84 to 1.54 for coronary disease, from 0.69 to 1.53 for stroke, and from 0.61 to 1.58 for peripheral arterial disease. In occupational populations, relative risks ranged from 0.40 to 2.14 for coronary disease mortality and from 0.30 to 1.33 for stroke mortality. Methodologic limitations, however, limited interpretation of the moderate-to-strong associations between high arsenic exposure and cardiovascular outcomes in Taiwan. In other populations or in occupational settings, the evidence was inconclusive. Because of the high prevalence of arsenic exposure, carefully performed studies of arsenic and cardiovascular outcomes should be a research priority. PMID- 16269586 TI - Determining lifestyle correlates of body mass index using multilevel analyses: the Tromso Study, 1979-2001. AB - Increases in overweight and obesity have been observed globally in both developed and developing countries. The authors assessed the relation between lifestyle factors and body mass index (BMI) (weight (kg)/height (m)2) in a population-based longitudinal study, using BMI and its subsequent change as responses in a multilevel model. The authors included 11,115 men and women aged 20-61 years at baseline who were living in the municipality of Tromso, Norway, and who participated in three or four consecutive health surveys between 1979-1980 and 2001. Baseline age, physical activity at work, coffee consumption, and desired BMI (i.e., the BMI that the subjects reported they would like to have) were positively associated with baseline BMI, whereas height, alcohol consumption, leisure-time physical activity, and level of education were inversely associated. Most relations were found to be stronger in women than in men. Clinically relevant effect sizes were observed for most of the significant associations, especially in women. For instance, on an ordinal scale, a one-category increase in educational level would decrease the mean baseline BMI among women by 0.30 kg/m2. Significant associations between several lifestyle factors and subsequent BMI change revealed that observed baseline associations were strengthened over time, especially in women. PMID- 16269587 TI - NEC: some coincidences are more equivalent than others. PMID- 16269588 TI - Fourier temporal interpolation improves electrocardiograph-gated myocardial perfusion SPECT. AB - The optimal temporal sampling rate in electrocardiograph-gated myocardial SPECT is questionable: low rates, typically 8 frames per cardiac beat (8fr/cb), favor image quality, whereas high rates, typically 16 frames per cardiac beat (16fr/cb), favor the accuracy of left ventricular (LV) functional parameters. We examined whether Fourier temporal interpolation (FTI) from 8fr/cb to 16fr/cb can combine the advantages of low and high rates. METHODS: In 34 patients imaged after stress injection of (99m)Tc-sestamibi, 4 sets of reconstructed gated slices were compared: a raw 16fr/cb acquisition (R16), a raw 8fr/cb acquisition (R8), a 16fr/cb set obtained by FTI of 8fr/cb projections (IP), and a 16fr/cb set obtained by FTI of 8fr/cb reconstructed slices (IS). LV ejection fraction (LVEF), end-diastolic volume (EDV), and end-systolic volume (ESV) obtained from the final LV volume curve were compared for the 4 datasets. Deviation of the whole LV volume curve was quantified for IP and IS with respect to R16. Image quality was evaluated by consensus reading of end-diastolic slices of the 4 sets. For R16, IP, and IS, cine display fluidity was quantified by a roughness index calculated from the LV volume curve. RESULTS: No differences in EDVs or ESVs were found among R16, IP, and IS, whereas R8 gave smaller EDVs and larger ESVs. LVEF was lower with R8, IP, and IS than with R16: -3.9%, -1.2%, and -1.3%, respectively. The LV volume curve was closer to R16 with IP than with IS. Image quality was better with IP and IS than with R8 and better with R8 than with R16. Cine display fluidity was better with IP than with R16 and better with R16 than with IS. CONCLUSION: FTI improved image quality not only over that provided by R16 but even over that provided by R8. The sole worsened LV functional parameter was LVEF, which was slightly underestimated with respect to that estimated by R16. Of the 2 FTI variants, IP was superior to IS for cine display fluidity and accuracy of the LV volume curve with respect to the data obtained with R16. Therefore, FTI to 16fr/cb performed before reconstruction on a pixel-by-pixel basis on 8fr/cb projections improves image quality and cine display fluidity over those of both R8 and R16 acquisitions at the sole cost of a 1% underestimation of LVEF. PMID- 16269589 TI - Cardiac and extracardiac sympathetic denervation in Parkinson's disease with orthostatic hypotension and in pure autonomic failure. AB - The uptake of 6-(18)F-fluorodopamine by cardiac noradrenergic nerves enables visualization of the sympathetic innervation of the left ventricular myocardium by PET. Patients with Parkinson's disease (PD) and orthostatic hypotension (OH) (PD+OH) or with pure autonomic failure (PAF) have markedly decreased myocardial 6 (18)F-fluorodopamine-derived radioactivity, consistent with cardiac sympathetic denervation, a phenomenon that neurochemical, neuropharmacologic, and, most recently, postmortem neuropathologic studies have confirmed. In this study, we examined whether 6-(18)F-fluorodopamine can visualize sympathetic innervation in extracardiac organs and, if so, whether patients with PD+OH or PAF have neuroimaging evidence of extracardiac noradrenergic denervation. METHODS: To validate the method, healthy volunteers underwent 6-(18)F-fluorodopamine scanning of the head, thorax, and abdomen, with or without treatment with desipramine to block sympathoneural uptake of catecholamines. (13)N-Ammonia scanning was used to address possible group differences in 6-(18)F-fluorodopamine delivery by blood perfusion. RESULTS: Desipramine treatment was associated with decreased 6-(18)F fluorodopamine-derived radioactivity in the heart, renal cortex, and thyroid gland but not in the liver, spleen, renal pelvis, or salivary glands. Both the PD+OH group and the PAF group had decreased 6-(18)F-fluorodopamine-derived radioactivity in the heart (P < 0.0001) and renal cortex (P = 0.02 and P = 0.005, respectively). The PD+OH group also had decreased radioactivity in the thyroid gland (P = 0.01). Neither group had decreased radioactivity in the other organs, after correction for (13)N-ammonia-derived radioactivity. CONCLUSION: 6-(18)F Fluorodopamine scanning visualizes sympathetic innervation in the heart, renal cortex, and thyroid gland. Both PD+OH and PAF involve decreased noradrenergic innervation that is most prominent in the heart but is also detectable in extracardiac organs. PMID- 16269590 TI - Relationship between residual blood flow in the infarct-related artery and scintigraphic infarct size, myocardial salvage, and functional recovery in patients with acute myocardial infarction. AB - In patients with acute myocardial infarction (AMI) before primary coronary stenting with adjunct glycoprotein IIb/IIIa receptor blockade, whether residual blood flow in the infarct-related artery (IRA) affects infarct size or myocardial salvage is not known. METHODS: This study included 118 patients with ST-segment elevation AMI who received coronary stenting plus abciximab. SPECT studies were performed before and 7-14 d after stenting. RESULTS: Patients were divided into a group with initial Thrombolysis in Myocardial Infarction (TIMI) flow grade < or = 1 (77 patients) and a group with initial TIMI flow grade > 1 (41 patients). The initial median perfusion defect and (in brackets) the 25th and 75th percentiles were 29.1% [21.0%; 52.0%] of the left ventricle in patients with TIMI flow grade < or = 1, versus 16.5% [8.0%; 33.1%] of the left ventricle in patients with TIMI flow grade > 1 (P < 0.001). Baseline left ventricular ejection fraction (54.0% [45.0; 63.0] vs. 57.0% [40.0; 62.0], P = 0.623) or extension of hypokinetic region (28.0 [14.0; 41.0] hypokinetic chords vs. 24.0 [13.0; 39.0] hypokinetic chords, P = 0.643) did not differ significantly between the group with TIMI flow grade < or = 1 and the group with TIMI flow grade > 1. Final infarct size was 11.0% [6.1%; 23.5%] of the left ventricle in the group with TIMI flow grade < = 1, versus 6.0% [2.0%; 12.8%] of the left ventricle in the group with TIMI flow > 1 (P = 0.008). Salvage index was 0.58 [0.38; 0.76] in the group with TIMI flow grade < or = 1, versus 0.61 [0.36; 0.74] in the group with TIMI flow grade > 1 (P = 0.952). At the day 14 angiography, patients with TIMI flow grade > 1 had better left ventricular ejection fraction (61.0% [54.0%; 68.0%] vs. 56.5% [42.9%; 65.0%]; P = 0.03) and a smaller hypokinetic region (7 chords [0; 22.0] vs. 16 chords [2.5; 30.0]; P = 0.024) than did patients with TIMI flow grade < or = 1. CONCLUSION: Preserved blood flow in the IRA in patients with AMI is associated with a smaller area at risk, a smaller infarct, and better recovery of regional and global left ventricular function. The proportion of initial area at risk salvaged by coronary stenting does not seem to depend on residual blood flow in the IRA. PMID- 16269591 TI - OSEM reconstruction, associated with temporal fourier and depth-dependant resolution recovery filtering, enhances results from sestamibi and 201Tl 16 interval gated SPECT. AB - Gated SPECT recorded with 16 intervals determines left ventricular (LV) ejection fraction more accurately than does gated SPECT recorded with 8 intervals but produces higher image noise. This study aimed to assess the results from sestamibi and (201)Tl 16-interval gated SPECT when both signal-to-noise ratio and spatial resolution were enhanced with an original method of reconstruction. METHODS: Forty patients with coronary artery disease underwent (201)Tl and sestamibi 16-interval gated SPECT and, to be used as a reference, cardiac MRI. Assessments of global and regional LV function provided by ordered-subsets expectation maximization (OSEM) with depth-dependant resolution recovery and temporal Fourier filtering were compared with those from conventional filtered backprojection (FBP) previously optimized by screening various filter frequencies and various temporal smoothing levels. RESULTS: For both tracers, LV ejection fraction was determined best when the association of OSEM with depth-dependant resolution recovery was used alone, with temporal Fourier filtering, or with a slight 2-frame temporal smoothing: Mean absolute values of relative errors ranged from 3.2% to 3.6% (4.0%-7.9% for FBP), and coefficient correlation ranged from 0.91 to 0.93 (0.70-0.91 for FBP). Among these 3 reconstruction methods, the association of OSEM with depth-dependant resolution recovery with temporal Fourier filtering provided the highest signal-to-noise ratio, with mean increases of 54% for sestamibi and 80% for (201)Tl when compared with FBP, and the best analysis of segmental contractility, with exact agreement rates with MRI being 73% for (201)Tl and 79% for sestamibi. CONCLUSION: OSEM associated with temporal Fourier filtering and depth-dependant resolution recovery filtering enhances the LV function assessment provided by sestamibi and (201)Tl 16-interval gated SPECT and dramatically reduces image noise, a property that enhances and facilitates image interpretation. PMID- 16269592 TI - Effects of carvedilol on myocardial sympathetic innervation in patients with chronic heart failure. AB - Carvedilol is a beta-blocking agent with antioxidant properties that has been shown to improve survival in chronic heart failure (CHF). Previous open-label studies have suggested that its use may have positive effects on the abnormalities of cardiac sympathetic innervation integrity and functioning. The present study aimed to test the hypothesis that carvedilol exerts its beneficial effects on hemodynamics in parallel with an action on myocardial sympathetic activity and with its antioxidant property. METHODS: A randomized, multicenter, double-blind, placebo-controlled study of carvedilol was conducted on 64 CHF patients. Patients underwent-before and after 6 mo of therapy with either carvedilol or placebo-measurements of cardiac sympathetic activity, circulating catecholamine level, and hemodynamic indices. Myocardial meta-(123)I iodobenzylguanidine ((123)I-MIBG) uptake was used to assess the changes in myocardial sympathetic activity. The antioxidant properties of the plasma were assessed by measuring the percentage of nonhemolyzed erythrocytes and the volume of plasma capable of inhibiting 50% of hemolysis after an oxidative stress. Echographic left ventricular (LV) diameters, radionuclide LV ejection fraction (LVEF), and exercise cardiopulmonary capacity were measured to evaluate the hemodynamic response. RESULTS: End-diastolic and end-systolic LV diameters decreased (both P < 0.05) and LVEF increased (P = 0.03) in the carvedilol group, whereas these parameters remained unchanged in the placebo group. Carvedilol did not alter the submaximal exercise cardiopulmonary capacity or the circulating catecholamine level. The beneficial hemodynamic effects in the carvedilol group were associated with an increase in myocardial (123)I-MIBG uptake as assessed by both planar and tomographic imaging (P < 0.01). Carvedilol had no detectable effect on antioxidant properties of the plasma. CONCLUSION: The benefits of carvedilol on resting hemodynamics appear to be associated with a partial recovery of cardiac adrenergic innervation functioning without detectable antioxidant effect in the plasma. PMID- 16269593 TI - Is iterative reconstruction an alternative to filtered backprojection in routine processing of dopamine transporter SPECT studies? AB - In general, striatal dopamine transporter (DAT) binding is assessed by use of data reconstructed by filtered backprojection (FBP). The aim of this study was to investigate whether the use of an iterative reconstruction algorithm (ordered subset expectation maximization [OSEM]) may provide results comparable to or even better than those obtained by standard FBP. METHODS: In 50 patients with parkinsonian syndromes, SPECT scans were acquired 4 h after injection of 185 MBq of (123)I-fluoropropyl-2beta-carbomethoxy-3beta-(4-iodophenyl)tropane ((123)I-FP CIT) by use of a triple-head gamma-camera fitted with low-energy, high-resolution fanbeam collimators. After reconstruction by FBP and OSEM, data were filtered with a Butterworth filter and corrected for attenuation. Patient studies were automatically fitted to a mean template with a corresponding 3-dimensional (3D) volume-of-interest map covering the striatum, caudate, and putamen as well as an occipital reference region to calculate specific DAT binding. In addition, studies with an anthropomorphic 3D striatal phantom were performed to mimic different pathologies. RESULTS: Visual assessment of phantom and patient data suggested a better separation between the caudate and the putamen in studies reconstructed by OSEM than in those reconstructed by FBP. There was an excellent correlation between specific DAT binding assessed by OSEM and that assessed by FBP (R(2) values: striatum, 0.999; caudate, 0.998; putamen, 0.998). Mean specific striatal binding obtained by OSEM was approximately 6% lower than that obtained by FBP. In no case was diagnostic information from OSEM inferior to that from FBP. CONCLUSION: Iterative reconstruction of (123)I-FP-CIT SPECT studies for the assessment of DAT is feasible in routine clinical practice. A close correlation between FBP and OSEM data suggested that the latter also allow reliable quantification of DAT binding. Because of a better separation between the caudate and the putamen in the visual evaluation, as suggested by phantom and patient studies, OSEM may even be considered the preferable approach. PMID- 16269594 TI - The role of imaging with (111)In-ibritumomab tiuxetan in the ibritumomab tiuxetan (zevalin) regimen: results from a Zevalin Imaging Registry. AB - The ibritumomab tiuxetan therapeutic regimen consists of a dose of rituximab, 250 mg/m(2), followed by (111)In-ibritumomab tiuxetan, for imaging, on day 1 and a dose of rituximab followed by (90)Y-ibritumomab tiuxetan, for therapy, on day 7, 8, or 9. Treatment with the Food and Drug Administration-approved regimen also requires that scans be performed at 2-24 h and at 48-72 h after the (111)In ibritumomab tiuxetan, with an optional third scan at 90-120 h, to confirm appropriate biodistribution. In the clinical trials before the approval of the regimen, only 1 patient (of approximately 400) was not treated with (90)Y ibritumomab tiuxetan after imaging with (111)In-ibritumomab tiuxetan, because of altered biodistribution. The Zevalin Imaging Registry was established by Biogen Idec Inc. to identify cases of potential altered biodistribution and to collect clinical information in cases in which the regimen was not completed after imaging. METHODS: The registry surveyed treating physicians to verify completion of treatment with the ibritumomab tiuxetan therapeutic regimen in patients treated with (111)In-ibritumomab tiuxetan between March 27, 2002, and March 31, 2003. RESULTS: Survey data were collected on 953 of an estimated 1,144-1,192 patients in whom ibritumomab tiuxetan therapy was initiated (case capture rate of 80%-83%). Thirty-eight cases were reported in which a decision not to treat was made after imaging with (111)In-ibritumomab tiuxetan (4.0% of all cases captured); 16 of these were for imaging reasons, and 22 were for medical reasons. Twelve of the 16 imaging cases met the criteria for altered biodistribution (1.3%). Of these 12 cases, 6 (0.6%) were suspected to be true altered biodistribution and 6 appeared to be due to the use of a procedure for radiolabeling (111)In-ibritumomab tiuxetan that differed from that in the prescribing information. All cases of altered biodistribution were seen on the first image (2-24 h) after the administration of (111)In-ibritumomab tiuxetan. The 22 cases in which decisions not to treat were made for medical reasons accounted for 2.3% of the cases. The majority of these cases (19/22) were in patients who had an expected biodistribution but had a rapid change in their clinical condition that precluded treatment. CONCLUSION: The rate of true altered biodistribution was 0.6% in the Zevalin Imaging Registry, which collected treatment decisions based on data from approximately 80% of all patients treated commercially in the first year after drug approval. All cases of altered biodistribution were apparent on the first image, obtained at 2-24 h after the administration of (111)In-ibritumomab tiuxetan. PMID- 16269595 TI - Potential of dual-time-point imaging to improve breast cancer diagnosis with (18)F-FDG PET. AB - The purpose of this study was to assess the utility of dual-time-point imaging for identifying malignant lesions in the breast by (18)F-FDG PET. METHODS: Fifty four breast cancer patients with 57 breast lesions underwent 2 sequential PET scans (dual-time-point imaging). The average percent change in standardized uptake values (SUVs) between time point 1 and time point 2 was calculated. All PET study results were correlated with follow-up surgical pathology results. RESULTS: Of the 57 breast lesions, 39 were invasive carcinoma and 18 were postbiopsy inflammation. Among the invasive carcinoma lesions, 33 (85%) showed an increase and 6 (15%) showed either no change or a decrease in SUVs over time. The percent change in SUVs from time point 1 to time point 2 (mean +/- SD) was +12.6% +/- 11.4% (P = 0.003). Of the 18 inflammatory lesions, 3 (17%) showed an increase and 15 (83%) showed either no change or a decrease in SUVs. The percent change in SUVs from time point 1 to time point 2 (mean +/- SD) was -10.2% +/- 16.5% (P = 0.03). Of the 57 normal contralateral breasts, 2 (3.5%) showed an increase and 55 (96.5%) showed either no change or a decrease in SUVs. The percent change in SUVs from time point 1 to time point 2 (mean +/- SD) was -15.8% +/- 17% (P = 0.005). CONCLUSION: There is increasing uptake of (18)F-FDG over time in breast malignancies, whereas the uptake of (18)F-FDG in inflammatory lesions and normal breast tissues decreases over time. A percent change of +3.75 or more in SUVs over time is highly sensitive and specific in differentiating inflammatory lesions from malignant lesions. PMID- 16269596 TI - Optimizing injected dose in clinical PET by accurately modeling the counting-rate response functions specific to individual patient scans. AB - To optimize the injected dose of radiopharmaceutical in PET, one needs to know its relationship to some metric of data quality for individual patient scans, such as noise-equivalent counting rate (NECR). In this paper, we show how one may accurately model the clinical NECR response corresponding to specific patient scans much as if a counting-rate test had been performed on each patient. We apply this technique to patient data and show how it can lead to improved clinical scanning protocols. METHODS: True and random coincidence rates expressed as functions of an appropriate measurable system parameter such as the detector single-event rate have functional forms that are largely independent of the object being scanned. Thus, reference true and random response functions may be scaled directly to the specific counting rates measured on a clinical scan, thereby yielding a curve of NECR versus injected dose. We have applied this technique to 2 groups of 163 clinical (18)F-FDG scans each. One of the groups was obtained on a lutetium oxyorthosilicate PET/CT scanner with conventional front end electronics, and the other was obtained on a lutetium oxyorthosilicate PET/CT scanner with a new digital data processing system (Pico-3D). RESULTS: At 90%-95% of maximum signal-to-noise ratio (SNR), the mean optimal dose for a 60-min uptake period ranged from 366 to 717 MBq depending on the electronics and randoms processing method. There was only a slight (1 MBq/kg) dependence of optimal dose on patient weight but a larger dependence on position in the body. Pico-3D electronics improved optimal data SNR by 35% for a 70-kg person, but in both cases NECR fell rapidly with increasing weight (1.4%/kg). For an equivalent data SNR, a 120-kg person would have to be scanned 2.3 times longer than a 60-kg person. Over this range of weight, the mean scatter fraction increased by 12% whereas the ratio of mean randoms to trues increased by 48%. CONCLUSION: The methodology developed here allows one to directly estimate the optimal dose to inject for specific clinical scans and permits a detailed analysis of the sources of noise in PET data and of their variation with parameters such as patient weight. PMID- 16269597 TI - Combined PET and microdialysis for in vivo assessment of intracellular drug pharmacokinetics in humans. AB - Because many drugs possess an intracellular site of action, the knowledge of intracellular concentration-time profiles is desirable. In the present study, PET, which measures total (i.e., intracellular, extracellular, and intravascular) concentrations of radiolabeled drugs in tissue, and microdialysis, which determines unbound drug concentrations in the extracellular space fluid of tissue, were combined to describe the intracellular pharmacokinetics of a model compound--that is, the (18)F-labeled antibiotic (18)F-ciprofloxacin--in vivo in humans. METHODS: Ten healthy male volunteers received a mixture of 687 +/- 50 MBq of (18)F-ciprofloxacin and 200 mg of unlabeled ciprofloxacin as an intravenous bolus infusion over 10 min. The pharmacokinetics of ciprofloxacin in skeletal muscle tissue were assessed by means of combined PET and in vivo microdialysis for 5 h after drug administration. A 3-compartment pharmacokinetic model was fitted to the tissue concentration-time profiles of ciprofloxacin measured by PET to estimate the rate constants of ciprofloxacin uptake and transport. RESULTS: In muscle tissue, mean total and extracellular peak concentration (C(max)) values of ciprofloxacin of 1.8 +/- 0.4 microg/mL and 0.7 +/- 0.2 microg/mL were attained at 95 +/- 34 min and 48 +/- 20 min after drug administration, respectively. The extracellular-to-intracellular exchange appeared to be very fast, with an estimated rate constant k(3) of 1.69 +/- 0.25 min(-1). An intracellular-to extracellular concentration ratio (C(intra)/C(extra)) of 3.2 +/- 0.8 was reached at 110 min after injection and followed by sustained intracellular retention of the antibiotic for the remainder of the experiment. The predicted extracellular concentration-time profiles from the compartmental modeling were in good agreement with the measured microdialysis data. CONCLUSION: The results obtained in the present study were in accordance with previous in vitro data describing cellular ciprofloxacin uptake and retention. The presently used PET/microdialysis combination might be useful during research and development of new drugs, for which knowledge of intracellular concentrations is of interest. PMID- 16269598 TI - Oncologic imaging in gynecologic malignancies. AB - (18)F-FDG PET imaging is underutilized in patients with gynecologic malignancies, and its role in current clinical practice has yet to be established. In cervical cancer, it has high sensitivity in detection of nodal disease. Its use is probably most suitable in patients with negative or ambiguous findings on other types of radiographic imaging. Data support its usefulness in asymptomatic cervical cancer patients with high tumor markers and negative conventional imaging findings, although more data are needed to ascertain whether it has a positive impact on survival. Similarly, its role in monitoring response to therapies needs to be consolidated. In ovarian cancer, (18)F-FDG PET holds promise in evaluation of recurrent or residual disease when other radiographic data are uncertain. In endometrial cancer, there are encouraging, although limited, data supporting the use of (18)F-FDG PET in patients with recurrent disease. To reduce extensive lymph node dissection in patients and to decrease subsequent morbidity, investigators have advocated applying the sentinel node technique to patients with cervical, endometrial, or vulvar cancers. The overall results are encouraging for the use of LS in planning surgical procedures, although more data and larger planned studies are needed to establish clinical utility in the surgical management of patients with these malignancies. PMID- 16269599 TI - Reproducibility of 3'-deoxy-3'-(18)F-fluorothymidine microPET studies in tumor xenografts in mice. AB - 3'-Deoxy-3'-(18)F-fluorothymidine ((18)F-FLT) has been used to image tumor proliferation in preclinical and clinical studies. Serial microPET studies may be useful for monitoring therapy response or for drug screening; however, the reproducibility of serial scans has not been determined. The purpose of this study was to determine the reproducibility of (18)F-FLT microPET studies. METHODS: C6 rat glioma xenografts were implanted into nude mice (n = 9) and grown to mean diameters of 5-17 mm for approximately 2 wk. A 10-min acquisition was performed on a microPET scanner approximately 1 h after (18)F-FLT (1.9-7.4 MBq [50-200 muCi]) was injected via the tail vein. A second microPET scan was performed approximately 6 h later on the same day after reinjection of (18)F-FLT to assess for reproducibility. Most of the mice were studied twice within the same week (for a total of 17 studies). Images were analyzed by drawing an ellipsoidal region of interest (ROI) around the tumor xenograft activity. Percentage injected dose per gram (%ID/g) values were calculated from the mean activity in the ROIs. Coefficients of variation and differences in %ID/g values between studies from the same day were calculated to determine the reproducibility after subtraction of the estimated residual tumor activity from the first (18)F-FLT injection. RESULTS: The coefficient of variation (mean +/- SD) for %ID/g values between (18)F-FLT microPET scans performed 6 h apart on the same day was 14% +/- 10%. The difference in %ID/g values between scans was -0.06% +/- 1.3%. Serum thymidine levels were mildly correlated with %ID/g values (R(2) = 0.40). Tumor size, mouse body weight, injected dose, and fasting state did not contribute to the variability of the scans; however, consistent scanning parameters were necessary to ensure accurate studies, in particular, controlling body temperature, the time of imaging after injection, and the ROI size. CONCLUSION: (18)F-FLT microPET mouse tumor xenograft studies are reproducible with moderately low variability. Serial studies may be performed to assess for significant changes in therapy response or for preclinical drug development. PMID- 16269600 TI - Evaluation of pharmacokinetics of 4-borono-2-(18)F-fluoro-L-phenylalanine for boron neutron capture therapy in a glioma-bearing rat model with hyperosmolar blood-brain barrier disruption. AB - This study evaluated the pharmacokinetics and biodistribution of 4-borono-2-(18)F fluoro-l-phenylalanine ((18)F-FBPA) after intracarotid injection and with blood brain barrier disruption (BBB-D) in F98 glioma-bearing F344 rats. The pharmacokinetics of l-p-boronophenylalanine (BPA) and (18)F-FBPA following different administration routes were compared to demonstrate the optimal delivery route and the time period for thermal neutron irradiation. METHODS: F98 glioma bearing rats were injected intravenously or intracarotidly with (18)F-FBPA and BPA and with or without mannitol-induced hyperosmotic BBB-D. The boron concentration and (18)F radioactivity in tissues were determined by invasive (inductively coupled plasma mass spectroscopy, gamma-counting) and noninvasive PET methods. RESULTS: The biodistributions of (18)F-FBPA and BPA in F98 glioma bearing rats were similar after intracarotid administration with BBB-D. The accumulation of BPA and (18)F-FBPA in brain tumor and the tumor-to-ipsilateral brain ratios were the highest after intracarotid injection with BBB-D, whereas the retention of boron drugs in contralateral brains exhibited only nonsignificant differences compared with those after intracarotid injection without BBB-D and intravenous injection. The high boron concentration in brain tumor (76.6 mug/g) and the high tumor-to-ipsilateral brain ratio (6.3) may afford enough radiation doses to destroy the tumor cells while sparing the normal tissues in boron neutron capture therapy. The pharmacokinetic parameters of k(el), k(12), k(21), and V(1) for intracarotid injection of (18)F-FBPA with BBB-D derived from the open 2-compartment model are 0.0206 +/- 0.0018 min(-1), 0.0260 +/- 0.0016 min(-1), 0.0039 +/- 0.0003 min(-1), and 3.1 +/- 0.1 mL, respectively. The effect of BBB-D varied depending on the anesthetic agents used and the anesthetic conditions. A smaller degree of BBB-D and, thus, lower boron concentrations in tumor and ipsilateral brain were observed under isoflurane anesthesia than under ketamine anesthesia. The k(12)/k(21) ratio may serve as a good indication for evaluating the extent of BBB-D, tumor uptake, and tumor-to brain ratio after intracarotid injection of boron compounds. CONCLUSION: Our findings provide important information for establishing an optimal treatment protocol when intracarotid injection with BPA after BBB-D is applied in clinical boron neutron capture therapy. PMID- 16269601 TI - Estimation of paclitaxel biodistribution and uptake in human-derived xenografts in vivo with (18)F-fluoropaclitaxel. AB - Paclitaxel (PAC) is widely used as a chemotherapy drug in the treatment of various malignancies, including breast, ovarian, and lung cancers. We examined the biodistribution of (18)F-fluoropaclitaxel ((18)F-FPAC) in mice with and without human breast cancer tumor xenografts by use of small-animal-dedicated PET (microPET) and clinically practical semiquantitative methods. We compared the PET data to data derived from direct harvesting and analysis of blood, organs, and breast carcinoma xenografts. METHODS: PET data were acquired after tail vein injection of (18)F-FPAC in nude mice. Tracer biodistribution in reconstructed images was quantified by region-of-interest analysis. Biodistribution also was assessed by harvesting and analysis of dissected organs, tumors, and blood after coadministration of (18)F-FPAC and (3)H-PAC. (18)F content in each tissue was assessed with a gamma-well counter, and (3)H content was quantified by scintillation counting of solubilized tissue after (18)F radioactive decay. RESULTS: The distributions of (18)F-FPAC and (3)H-PAC were very similar, with the highest concentrations in the small intestine, the lowest concentrations in the brain, and intermediate concentrations in tumor. Uptake in these and other tissues was not inhibited by the presence of more pharmacologic doses of unlabeled PAC. Administration of the P-glycoprotein modulator cyclosporine doubled the uptake of both (18)F-FPAC and (3)H-PAC into tumor. CONCLUSION: PET studies with (18)F-FPAC can be used in conjunction with clinically practical quantification methods to yield estimates of PAC uptake in breast cancer tumors and normal organs noninvasively. PMID- 16269602 TI - Evaluation of novel whole-body high-resolution rodent SPECT (Linoview) based on direct acquisition of linogram projections. AB - Studies of the biodistribution of radiolabeled compounds in rodents frequently are performed during the process of development of new pharmaceutical drugs. This article presents the evaluation of a new whole-body animal SPECT system, called the Linoview SPECT system. METHODS: Linoview SPECT is based on the linear orbit acquisition technique associated with slit-aperture collimators mounted on 4 pixelated CsI(Na) detectors composed of an array of small, individual crystal elements. Sliding iridium rods allow variation of the collimator aperture. Hot rod and cold-rod phantoms filled with (99m)Tc were imaged. Mice were imaged, and kidney radioactivity was measured after injection of (99m)Tc-dimercaptosuccinic acid and (111)In-diethylenetriaminepentaacetic acid-d-Phe(1)-octreotide ((111)In pentetreotide; Octreo-Scan(111)). RESULTS: Phantom studies showed that hot rods separated by 0.35 mm can be distinguished and that 0.65-mm-diameter cold rods can be visualized, both at low-counting-rate acquisitions (111 and 59 MBq x h, respectively). In both mouse studies, the SPECT images allowed a clear delineation of the radioactivity concentrated over the cortex area of the kidneys, whereas the pelvis and the pelviureteral junction (1 mm) appeared as cold areas. The quantitative data derived from SPECT were in good agreement with the radioactivity counting obtained with a gamma-counter after isolation of the kidneys. In addition, in the mouse injected with (111)In-pentetreotide, the kidney radioactivity distribution seen with SPECT was in agreement with the ex vivo autoradiograms of the isolated kidneys. CONCLUSION: The phantom studies showed a clear improvement of the spatial resolution over the results reported in the literature with other dedicated small-animal SPECT systems, especially in cold-rod phantom studies. The increased performance can be ascribed to the high stability of the system with regard to the statistical noise present in the acquired data. The mouse studies showed that this system will be most useful for in vivo high-resolution SPECT and quantitative biodistribution studies in rodents, even with medium-energy radioisotopes that are difficult to image, such as (111)In. PMID- 16269603 TI - Preparation and evaluation of (68)Ga-DOTA-hEGF for visualization of EGFR expression in malignant tumors. AB - Detection of epidermal growth factor receptor (EGFR) overexpression in many carcinomas provides important diagnostic information, which can influence patient management. The use of PET may enable such detection in vivo by a noninvasive procedure with high sensitivity. The aim of this study was to develop a method for preparation of a positron-emitting tracer based on a natural ligand to EGFR, the recombinant human epidermal growth factor (hEGF), and to perform a preclinical evaluation of the tracer. METHODS: DOTA-hEGF (DOTA is 1,4,7,10 tetraazacyclododecane-N,N',N'',N'''-tetraacetic acid) was prepared by coupling of a N-sulfosuccinimide ester of DOTA to hEGF. The conjugate was labeled with a generator-produced positron-emitting nuclide, (68)Ga (half-life = 68 min), using microwave heating. Binding specificity, affinity, internalization, and retention of (68)Ga-DOTA-hEGF was studied in 2 EGFR-expressing cell lines, U343 glioma cells and A431 cervical carcinoma cells. Biodistribution and microPET visualization studies were performed in BALB/c nu/nu mice bearing A431 carcinoma xenografts. RESULTS: A 1-min-long microwave-assisted labeling provided radioactivity incorporation of 77% +/- 4%. Both cell lines demonstrated receptor specific uptake of the conjugate, rapid internalization of the tracer, and good retention of radioactivity. Binding to both cell lines occurred with high affinity, approximately 2 nmol/L. The biodistribution study demonstrated accumulation of radioactivity in xenografts and in EGFR-expressing organs. The microPET imaging study enabled visualization of tumors and demonstrated quick- within 5 min--localization of radioactivity in tumors. CONCLUSION: (68)Ga-DOTA hEGF has potential for imaging EGFR overexpression in tumors. PMID- 16269604 TI - MicroPET imaging of gene transfer with a somatostatin receptor-based reporter gene and (94m)Tc-Demotate 1. AB - Gene therapy trials would benefit greatly from the use of noninvasive imaging to determine the location, magnitude, and time course of gene transfer. Somatostatin receptor subtype 2 (SSTR2) has been used as a reporter probe for gamma-camera imaging of gene transfer in animal models. PET has greater sensitivity than gamma camera imaging and therefore would have an advantage for the imaging of SSTR2 gene transfer. METHODS: An adenovirus (AdHASSTR2) carrying sstr2, which encodes an N-terminal hemagglutinin epitope, was used for evaluating SSTR2 gene transfer. The somatostatin analog Demotate 1 (Tyr(3)-octreotate conjugated to the 1,4,8,11 tetraazaundecane chelator) was used for chelation of the positron emitter (94m)Tc (half-life, 52 min) and targeting to SSTR2. Gene transfer was evaluated in vitro with A-427 non-small cell lung cancer cells after infection with AdHASSTR2 by (94m)Tc-Demotate 1 binding and internalization assays. In vivo biodistribution and microPET studies were conducted with mice bearing A-427 tumor xenografts directly injected with AdHASSTR2 to determine the tumor localization of (94m)Tc Demotate 1. RESULTS: (94m)Tc-Demotate 1 bound with high affinity and was internalized rapidly into AdHASSTR2-infected A-427 cells. Biodistribution studies showed uptake of (94m)Tc-Demotate 1 in tumors infected with AdHASSTR2 (4.0 percentage injected dose per gram [%ID/g] at 2 h) and background uptake in tumors infected with a control adenovirus (0.8 %ID/g at 2 h). The uptake of (94m)Tc Demotate 1 in AdHASSTR2-infected tumors was greater than the uptake in all other tissues, except for the kidneys and the SSTR2-positive pancreas. MicroPET imaging showed similar results, with clear uptake of (94m)Tc-Demotate 1 in AdHASSTR2 infected tumors, background uptake in control tumors, and clearance through the kidneys. CONCLUSION: These studies show that the positron-emitting somatostatin analog (94m)Tc-Demotate 1 could be used to determine SSTR2 gene transfer by microPET imaging, a result that will improve the sensitivity of the SSTR2 reporter gene system. PMID- 16269605 TI - (89)Zr as a PET surrogate radioisotope for scouting biodistribution of the therapeutic radiometals (90)Y and (177)Lu in tumor-bearing nude mice after coupling to the internalizing antibody cetuximab. AB - Immuno-PET as a scouting procedure before radioimmunotherapy (RIT) aims at confirming tumor targeting and accurately estimating radiation dose delivery to both tumor and normal tissues and might therefore be of value for selection of patient candidates for RIT. A prerequisite for this approach is that PET radioimmunoconjugates and RIT radioimmunoconjugates must show a similar biodistribution. In the present study, we evaluated the potential of the long lived positron emitter (89)Zr to predict biodistribution of the residualizing therapeutic radiometals (88)Y (as a substitute for (90)Y) and (177)Lu when labeled to the monoclonal antibody (mAb) cetuximab via different types of chelates. Cetuximab was selected as a model mAb because it abundantly internalizes after binding to the epidermal growth factor receptor. METHODS: Cetuximab was labeled with (89)Zr using succinylated desferrioxamine B (N-sucDf). The chelates p-benzyl isothiocyanate-1,4,7,10-tetraazacyclododecane-1,4,7, 10 tetraacetic acid (p-SCN-Bz-DOTA) and p-isothiocyanatobenzyl diethylenetriaminepentaacetic acid (p-SCN-Bz-DTPA) were both used for radiolabeling with (88)Y and (177)Lu. For measurement of the in vitro stability of each of the 5 radioimmunoconjugates, samples were incubated in freshly prepared human serum at 37 degrees C up to 16 d. Biodistribution was assessed at 24, 48, 72, and 144 h after intraperitoneal coinjection of the PET and RIT conjugates in nude mice bearing the squamous cell carcinoma xenograft line A431. RESULTS: Cetuximab premodification with N-sucDf, p-SCN-Bz-DOTA, or p-SCN-Bz-DTPA resulted in chelate-to-mAb molar ratios of about 1. After radiolabeling and purification, the radiochemical purity and immunoreactive fraction of the conjugates always exceeded 97% and 93%, respectively. All conjugates were stable in serum, showing a radioactivity release of less than 5% until day 7. From day 7 until day 16, an enhanced release was observed for the (89)Zr-N-sucDf, (88)Y-p SCN-Bz-DTPA, and (177)Lu-p-SCN-Bz-DTPA conjugates. The coinjected PET and RIT conjugates showed similar biodistributions, except for the thighbone and sternum. For example, the (89)Zr-N-sucDf conjugate showed a 2.0-2.5 times higher radioactivity accretion in the thighbone than did the RIT conjugates at 72 h after injection. CONCLUSION: In view of the advantages of PET over SPECT, (89)Zr immuno-PET is a promising modality for in vivo scouting of (90)Y- and (177)Lu labeled mAbs, although care should be taken when estimating bone marrow doses. PMID- 16269607 TI - Biodistribution and radiation dosimetry estimates of 1-(2'-deoxy-2'-(18)F-Fluoro 1-beta-D-arabinofuranosyl)-5-bromouracil: PET imaging studies in dogs. AB - This study reports on the biodistribution and radiation estimates of 1-(2'-deoxy 2'-(18)F-fluoro-1-beta-d-arabinofuranosyl)-5-bromouracil ((18)F-FBAU), a potential tracer for imaging DNA synthesis. METHODS: Three normal dogs were intravenously administered (18)F-FBAU and a dynamic PET scan was performed for 60 min over the upper abdomen followed by a whole-body scan for a total of 150 min. Blood samples were collected at stipulated time intervals to evaluate tracer clearance and metabolism. Tissue samples of various organs were analyzed for tracer uptake and DNA incorporation. Dynamic accumulation of the tracer in different organs was derived from reconstructed PET images. The radiation dosimetry of (18)F-FBAU was evaluated using the MIRD method. RESULTS: At 60 min after injection, blood analysis found >90% of the activity in unmetabolized form. At 2 h after injection, (18)F-FBAU uptake was highest in proliferating tissues (mean SUVs: marrow, 2.6; small intestine, 4.0), whereas nonproliferative tissues showed little uptake (mean SUVs: muscle, 0.75; lung, 0.70; heart, 0.85; liver, 1.28). Dynamic image analysis over 60 min showed progressive uptake of the tracer in marrow. Extraction studies demonstrated that most of the activity in proliferative tissues was in the acid-insoluble fraction (marrow, 83%; small intestine, 73%), consistent with incorporation into DNA. In nonproliferative tissue, most of the activity was not found in the acid-insoluble fraction (>84% for heart, muscle, and liver). CONCLUSION: These results demonstrate that (18)F FBAU was resistant to metabolism, readily incorporated into DNA in proliferating tissues, and showed good contrast between organs of variable DNA synthesis. These findings indicate that (18)F-FBAU may find use in measuring DNA synthesis with PET. PMID- 16269606 TI - Therapeutic efficacy and tumor dose estimations in radioimmunotherapy of intraperitoneally growing OVCAR-3 cells in nude mice with (211)At-labeled monoclonal antibody MX35. AB - The purpose of this study was to investigate the therapeutic efficacy of-and to estimate the absorbed dose to-tumor cells from radioimmunotherapy (RIT) in an ovarian cancer model using the alpha-particle-emitting nuclide (211)At labeled to monoclonal antibody (mAb) MX35. Previous studies on mAb MOv18 did not allow for dosimetry because of antigen shedding in vitro. METHODS: Five-week-old female nude BALB/c nu/nu mice were inoculated intraperitoneally with 1 x 10(7) cells of the human tumor cell line OVCAR-3. Three weeks later, the animals were given approximately 400, 800, or 1,200 kBq of (211)At-labeled mAb MX35 intraperitoneally. As controls, one group of animals was injected with unlabeled mAb and another group was injected with phosphate-buffered saline (PBS). Another group was given approximately 400 kBq of (211)At labeled to the previously investigated mAb MOv18 for efficacy comparison. Two months after treatment, the animals were sacrificed and the presence of macroscopic and microscopic tumors, as well as ascites, was determined. The absorbed dose to tumor cells on the peritoneal surface was estimated in terms of the sum of a specific and a nonspecific contribution. The specific contribution, arising from mAbs binding to the antigenic sites on the cell membrane, was calculated using a dynamic compartment model developed in-house and Monte Carlo software. The model used as input values the number of mAbs injected into the abdominal cavity, N(mAb), the specific activity, A(sp), the association rate constant, k(on), and the maximal number of mAbs bound per cell, B(max)-all determined by in vitro experiments. This specific component of the absorbed dose was calculated for assumed cell cluster sizes with radii of 25, 50, and 100 microm. The nonspecific contribution to the absorbed dose was derived from unbound mAbs freely circulating in the abdominal cavity, also using the Monte Carlo software. RESULTS: In the control groups given unlabeled MX35 or PBS, all 18 animals had ascites, 6 of 9 animals in each group had macroscopic tumors, and all animals had microscopic growth. In the 3 groups given different amounts of (211)At-MX35, only 3 of 25 animals developed ascites. None of these animals had any sign of macroscopic tumors, but 8 had microscopic growth. In the group given (211)At-MOv18, no animals had ascites or macroscopic tumors, but 3 of 10 animals had microscopic tumors. After injecting 400 kBq of (211)At-MX35, the absorbed dose due to specific binding, for a cell cluster with a radius of 50 microm, ranged from 413 to 223 Gy between 0- and 45 microm distance from the cluster center, assuming a homogeneous distribution of (211)At-MX35 in the cluster. The contribution from unbound (211)At-MX35 and (211)At-MX35 only distributed on the cluster surface, for this cluster size, ranged from 7 to 14 Gy and from 29 to 94 Gy, between 0- and 45-microm distance from the cluster center, respectively. The calculated total absorbed doses are in a clinically relevant range and were effective as verified in the nude mice with subclinical intraperitoneal growth of OVCAR-3 cells. CONCLUSION: (211)At-MX35 injected intraperitoneally exhibits a high efficacy when treating micrometastatic growth of the ovarian cancer cell line OVCAR-3 on the peritoneum of nude mice. PMID- 16269608 TI - Evaluation of (76)Br-FBAU as a PET reporter probe for HSV1-tk gene expression imaging using mouse models of human glioma. AB - The utility of 5-(76)Br-bromo-2'-fluoro-2'-deoxyuridine ((76)Br-FBAU), a uracil analog, as a PET reporter probe for use with the herpes simplex virus type 1 thymidine kinase (HSV1-tk) reporter gene system for gene expression imaging was evaluated in vivo and in vitro using human and rat glioma cells. METHODS: Human glioma cell lines U87 and U251 were transduced with replication-defective adenovirus constitutively expressing HSV1-tk (Ad.TK) or a control expressing green fluorescent protein (Ad.GFP). These cells were incubated with (76)Br-FBAU for 20-120 min to determine the percentage of total dose uptake. In vitro uptake of equimolar concentrations (1.8 x 10(-8) mol/L) of (76)Br-FBAU and 2'-fluoro-2' deoxy-5-iodouracil-beta-d-arabinofuranoside ((14)C-FIAU) was also determined in RG2-TK rat glioma cells stably expressing HSV1-tk and in control RG2 cells at 30 120 min. In vivo uptake of (76)Br-FBAU was determined in subcutaneous U87 tumor intratumorally transduced with Ad.TK by ex vivo biodistribution. Uptake in intracranial U87 tumors transduced with Ad.TK expressing HSV1-tk was measured by brain autoradiography. In vivo PET was performed on subcutaneous and intracranial U87 tumors transduced with Ad.TK and on subcutaneous and intracranial stably expressing RG2-TK tumors. RESULTS: U87 and U251 cells transduced with Ad.TK had significantly increased uptake of (76)Br-FBAU compared with cells transduced with Ad.GFP over 20-120 min. In stably expressing cells at 120 min, (14)C-FIAU uptake in RG2-TK tumor cells was 11.3 %ID (percentage injected dose) and in RG2 control cells was 1.7 %ID, and (76)Br-FBAU uptake in RG2-TK tumor cells was 14.2 %ID and in RG2 control cells was 1.5 %ID. Ex vivo biodistribution of subcutaneous U87 tumors transduced with Ad.TK accumulated (76)Br-FBAU significantly more than in the control Ad.GFP transduced tumor and normal tissue, with the lowest uptake in brain. Autoradiography showed localized uptake in intracranial U87 and U251 cells transduced with Ad.TK. PET image analyses of mice with RG2-TK tumors resulted in an increased tumor-to-background ratio of 13 and 26 from 2 to 6 h after injection, respectively, in intracranial tumors. CONCLUSION: (76)Br-FBAU accumulates in glioma cells constitutively expressing HSV1-tk by either adenoviral transduction or in stably expressing cell lines both in vitro and in vivo. (76)Br-FBAU shows promise as a PET reporter probe for use with the HSV1-tk in vivo gene expression imaging system. PMID- 16269609 TI - PET study of the neuroprotective effect of TRA-418, an antiplatelet agent, in a monkey model of stroke. AB - The aim of this study was to evaluate the neuroprotective effect of TRA-418, an antiplatelet agent, using PET in a monkey model of stroke. TRA-418 is a nonprostanoid compound with dual action: antagonistic effects on thromboxane A(2) receptors and agonistic effects on prostaglandin I(2) receptors. METHODS: Via a transorbital approach, cynomolgus monkeys underwent a 3-h occlusion of the right middle cerebral artery (MCA), followed by reperfusion and observation for 4 d. Starting 2 h after the MCA occlusion, TRA-418 was administered at low and high doses (6 animals at each dose). Six control animals received a bolus and infusion of drug vehicle after MCA occlusion. Steady-state (15)O continuous inhalation was used for assessment of local cerebral blood flow, cerebral metabolic rate of oxygen, and oxygen extraction fraction using high-resolution PET. Five consecutive PET scans (before occlusion; 2 h after occlusion; and 2 h, 24 h, and 4 d after reperfusion) were obtained for each monkey. The extent of the cerebral damage due to ischemia was measured histologically at 4 d after reperfusion. RESULTS: Histologic observation 4 d after MCA occlusion showed that cerebral damage was less (P = 0.05) in animals treated with high-dose TRA-418 than in control animals. Although not affecting cerebral blood flow during the experiments, treatment with TRA-418 significantly (P < 0.05) suppressed reduction of the cerebral metabolic rate of oxygen after reperfusion. CONCLUSION: Our observations suggest that TRA-418 has neuroprotective action, as displayed in a primate model of stroke using PET monitoring. PMID- 16269610 TI - A phase 1 trial of donor lymphocyte infusions expanded and activated ex vivo via CD3/CD28 costimulation. AB - Donor lymphocyte infusions (DLIs) induce potent graft versus tumor (GVT) effects for relapsed chronic myelogenous leukemia (CML) after allogeneic stem cell transplantation (SCT) but are disappointing for other diseases. Disease resistance can occur if donor T cells are not appropriately activated in vivo. Ex vivo T-cell activation might overcome disease-induced anergy and augment GVT activity. We performed a phase 1 trial of ex vivo-activated DLI (aDLI) for 18 patients with relapse after SCT. Activated donor T cells are produced through costimulation with anti-CD3- and anti-CD28-coated beads. Patients with aggressive malignancies received induction chemotherapy, and all patients received conventional DLI (median, 1.5 x 10(8) mononuclear cells/kg) followed 12 days later by aDLI. Activated DLI was dose escalated from 1 x 10(6) to 1 x 10(8) CD3+ cells per kilogram in 5 levels. Seven patients developed acute graft versus host disease (GVHD) (5 grade I-II, 2 grade III), and 4 developed chronic GVHD. Eight patients achieved complete remission, including 4 of 7 with acute lymphocytic leukemia (ALL), 2 of 4 with acute myelogenous leukemia (AML), 1 with chronic lymphocytic leukemia (CLL), and 1 of 2 with non-Hodgkin lymphoma (NHL). Four complete responders relapsed while 4 remain alive in remission a median 23 months after aDLI. Overall, 10 of 18 remain alive 11 to 53 months after aDLI. Adoptive transfer of costimulated activated allogeneic T cells is feasible, does not result in excessive GVHD, and may contribute to durable remissions in diseases where conventional DLI has been disappointing. PMID- 16269611 TI - CXCR4: a key receptor in the crosstalk between tumor cells and their microenvironment. AB - Signals from the microenvironment have a profound influence on the maintenance and/or progression of hematopoietic and epithelial cancers. Mesenchymal or marrow derived stromal cells, which constitute a large proportion of the non-neoplastic cells within the tumor microenvironment, constitutively secrete the chemokine stromal cell-derived factor-1 (SDF-1/CXCL12). CXCL12 secretion by stromal cells attracts cancer cells, acting through its cognate receptor, CXCR4, which is expressed by both hematopoietic and nonhematopoietic tumor cells. CXCR4 promotes tumor progression by direct and indirect mechanisms. First, CXCR4 is essential for metastatic spread to organs where CXCL12 is expressed, and thereby allows tumor cells to access cellular niches, such as the marrow, that favor tumor-cell survival and growth. Second, stromal-derived CXCL12 itself can stimulate survival and growth of neoplastic cells in a paracrine fashion. Third, CXCL12 can promote tumor angiogenesis by attracting endothelial cells to the tumor microenvironment. CXCR4 expression is a prognostic marker in various types of cancer, such as acute myelogenous leukemia or breast carcinoma. Promising results in preclinical tumor models indicate that CXCR4 antagonists may have antitumor activity in patients with various malignancies. Collectively, these observations reveal that CXCR4 is an important molecule involved in the spread and progression of a variety of different tumors. As such, CXCR4 antagonists, although initially developed for treatment of AIDS, actually may become effective agents for the treatment of neoplastic disease. PMID- 16269612 TI - CD43 is a ligand for E-selectin on CLA+ human T cells. AB - The recruitment of memory T cells from blood into tissues is a central element of immune surveillance and adaptive immune responses and a key feature of chronic cutaneous inflammatory diseases such as psoriasis and atopic dermatitis. Human memory T cells that infiltrate skin express the carbohydrate epitope cutaneous lymphocyte-associated antigen (CLA). Expression of the CLA epitope on T cells has been described on P-selectin glycoprotein ligand-1 (PSGL-1) and associated with the acquisition of both E-selectin and P-selectin ligand functions. In this report, we show that CD43, a sialomucin expressed constitutively on T cells, can also be decorated with the CLA epitope and serve as an E-selectin ligand. CLA expressed on CD43 was found exclusively on the high-molecular-weight (125 kDa) glycoform bearing core-2-branched O-linked glycans. CLA+ CD43 purified from human T cells supported tethering and rolling in shear flow via E-selectin but did not support binding of P-selectin. The identification and characterization of CD43 as a T-cell E-selectin ligand distinct from PSGL-1 expands the role of CD43 in the regulation of T-cell trafficking and provides new targets for the modulation of immune functions in skin. PMID- 16269613 TI - Remodeling specific immunity by use of MHC tetramers: demonstration in a graft versus-host disease model. AB - Major histocompatibility complex (MHC) molecules carrying selected peptides will bind specifically to their cognate T-cell receptor on individual clones of reactive T cells. Fluorescently labeled, tetrameric MHC-peptide complexes have been widely used to detect and quantitate antigen-specific T-cell populations via flow cytometry. We hypothesized that such MHC-peptide tetramers could also be used to selectively deplete unique reactive T-cell populations, while leaving the remaining T-cell repertoire and immune response intact. In this report, we successfully demonstrate that a tetramer-based depletion of T cells can be achieved in a murine model of allogeneic bone marrow transplantation. Depletion of a specific alloreactive population of donor splenocytes (< 0.5% of CD8+ T cells) prior to transplantation significantly decreased morbidity and mortality from graft-versus-host disease. There was no early regrowth of the antigen specific T cells in the recipient and in vivo T-cell proliferation was greatly reduced as well. Survival was increased more than 3-fold over controls, yet the inherent antitumor activity of the transplant was retained. This method also provides the proof-of-concept for similar strategies to selectively remove other unwanted T-cell clones, which could result in novel therapies for certain autoimmune disorders, T-cell malignancies, and solid organ graft rejection. PMID- 16269615 TI - AID expression identifies interfollicular large B cells as putative precursors of mature B-cell malignancies. AB - Neoplastic transformation of mature B cells can be triggered by class-switch recombination of the immunoglobulin gene, which aberrantly targets a protooncogene and promotes translocation. Class-switch recombination is initiated by the B-cell-specific protein activation-induced cytidine deaminase (AID). Using immunohistochemistry with a newly generated monoclonal antibody and quantitative reverse-transcription-polymerase chain reaction (RT-PCR) on microdissected tissue from lymph node, tonsil, and thymus, we demonstrate that AID expression is found in secondary lymphoid organs outside germinal centers and in the thymic medulla at substantial levels. This is accompanied by the presence of circle transcripts, indicating class-switch recombination to be active at these sites. The dominant AID-expressing cell population outside germinal centers displays cytomorphologic properties corresponding to those that define the recently characterized interfollicular large B-cell subset. These findings indicate that interfollicular large B cells and AID-expressing B lymphocytes of the thymic medulla could give rise to mature B-cell malignancies. PMID- 16269614 TI - Failure of HIV-exposed CD4+ T cells to activate dendritic cells is reversed by restoration of CD40/CD154 interactions. AB - Because interactions between activated CD4+ T cells and antigen-presenting cells (APCs) are crucial for optimal APC function, defective CD4+ T-cell activation may contribute to APC dysregulation in HIV infection. Here, we show that CD4+ T cells exposed during stimulation to noninfectious HIV having functional envelope glycoproteins failed to provide activation signals to autologous dendritic cells (DCs). Consequently, important DC functions, including production of immunoregulatory cytokines (interleukin-12 p40 and interleukin-10) and up regulation of costimulatory molecules (CD86, CD40, CD83), as well as the capacity to stimulate naive allogeneic T cells, were all adversely affected. The blunted up-regulation of CD154 in CD4+ T cells that were activated in the presence of noninfectious viruses is likely to be the major underlying mechanism for these defects. Addition of recombinant trimeric CD154 could restore production of cytokines by DCs cocultured with HIV-exposed T cells. Moreover, the functional defects mediated by coculture with HIV-exposed T cells were similar to those following antibody blockade of CD40-CD154 interactions. HIV-mediated blunted CD154 expression may thus play an important role in the suppression of cell mediated immunity seen in HIV infection. PMID- 16269616 TI - CD4+/CD8+ macrophages infiltrating at inflammatory sites: a population of monocytes/macrophages with a cytotoxic phenotype. AB - We found a population of nonlymphoid cells expressing both CD4 and CD8 in peripheral blood mononuclear cells (PBMCs) of human T-cell leukemia virus type-I pX transgenic rats with autoimmune diseases. These cells, which showed a monocytic phenotype, were also found in wild-type rats, and their number increased by adjuvant-assisted immunization. GM-CSF increased the number of these double-positive (DP) monocytes in PBMCs. Consistent with the idea that DP monocytes differentiate into DP macrophages at sites of inflammation, we found infiltration of DP macrophages at the site of myosin-induced myocarditis in wild type rats; these cells exhibited a T-helper 1 (Th1)-type cytokine/chemokine profile and expressed high levels of Fas ligand, perforin, granzyme B, and NKR-P2 (rat orthologue of human NKG2D). Adoptive transfer of GFP-positive spleen cells confirmed hematogenous origin of DP macrophages. DP monocytes had a cytotoxic phenotype similar to DP macrophages, indicating that this phenotypic specialization occurred before entry into a tissue. In line with this, DP monocytes killed tumor cells in vitro. Combined evidence indicates that certain inflammatory stimuli that induce GM-CSF trigger the expansion of a population of DP monocytes with a cytotoxic phenotype and that these cells differentiate into macrophages at inflammatory sites. Interestingly, human PBMCs also contain DP monocytes. PMID- 16269617 TI - Survival of the fittest: in vivo selection and stem cell gene therapy. AB - Stem cell gene therapy has long been limited by low gene transfer efficiency to hematopoietic stem cells. Recent years have witnessed clinical success in select diseases such as X-linked severe combined immunodeficiency (SCID) and ADA deficiency. Arguably, the single most important factor responsible for the increased efficacy of these recent protocols is the fact that the genetic correction provided a selective in vivo survival advantage. Since, for most diseases, there will be no selective advantage of gene-corrected cells, there has been a significant effort to arm vectors with a survival advantage. Two-gene vectors can be used to introduce the therapeutic gene and a selectable marker gene. Efficient in vivo selection strategies have been demonstrated in clinically relevant large-animal models. Mutant forms of the DNA repair-enzyme methylguanine methyltransferase in particular have allowed for efficient in vivo selection and have achieved sustained marking with virtually 100% gene-modified cells in large animals, and with clinically acceptable toxicity. Translation of these strategies to the clinical setting is imminent. Here, we review how in vivo selection strategies can be used to make stem cell gene therapy applicable to the treatment of a wider scope of genetic diseases and patients. PMID- 16269618 TI - Dynamic shifts in LFA-1 affinity regulate neutrophil rolling, arrest, and transmigration on inflamed endothelium. AB - Polymorphonuclear leukocyte (PMN) recruitment to vascular endothelium during acute inflammation involves cooperation between selectins, G-proteins, and beta2 integrins. LFA-1 (CD11a/CD18) affinity correlates with specific adhesion functions because a shift from low to intermediate affinity supports rolling on ICAM-1, whereas high affinity is associated with shear-resistant leukocyte arrest. We imaged PMN adhesion on cytokine-inflamed endothelium in a parallel plate flow chamber to define the dynamics of beta2-integrin function during recruitment and transmigration. After arrest on inflamed endothelium, high affinity LFA-1 aligned along the uropod-pseudopod major axis, which was essential for efficient neutrophil polarization and subsequent transmigration. An allosteric small molecule inhibitor targeted to the I-domain stabilized LFA-1 in an intermediate-affinity conformation, which supported neutrophil rolling but inhibited cell polarization and abrogated transmigration. We conclude that a shift in LFA-1 from intermediate to high affinity during the transition from rolling to arrest provides the contact-mediated signaling and guidance necessary for PMN transmigration on inflamed endothelium. PMID- 16269620 TI - CD11c+ dendritic cells and plasmacytoid DCs are activated by human cytomegalovirus and retain efficient T cell-stimulatory capability upon infection. AB - It has been suggested that human cytomegalovirus (HCMV) evades the immune system by infecting and paralyzing antigen-presenting cells. This view is based mainly on studies of dendritic cells (DCs) obtained after culture of monocytes (moDCs). It is contradicted by the asymptomatic course of HCMV infection in healthy persons, indicating that other key antigen-presenting cells induce an efficient immune response. Here we show that HCMV activates CD11c+ DCs and plasmacytoid DCs (PDCs). In contrast to moDCs, CD11c+ DCs and PDCs produced interferon (IFN) type 1 when exposed to HCMV. Autocrine IFN type 1 partially protected CD11c+ DCs against infection, whereas PDCs were resistant to HCMV even when IFN type 1 activity was inhibited. HCMV exposure induced the maturation of CD11c+ DCs by IFN type 1-dependent and -independent mechanisms. Importantly, CD11c+ DCs infected by inhibiting IFN type 1 activity retained full capacity to stimulate T cells. Renal transplant recipients receiving immunosuppressive treatment had lower frequencies of CD11c+ DCs and PDCs in blood than did healthy controls. The results show that HCMV activates the immune system by interacting with CD11c+ DCs and PDCs and that recipients of renal transplants have low frequencies of these cell types in blood. PMID- 16269619 TI - Selection based on CD133 and high aldehyde dehydrogenase activity isolates long term reconstituting human hematopoietic stem cells. AB - The development of novel cell-based therapies requires understanding of distinct human hematopoietic stem and progenitor cell populations. We recently isolated reconstituting hematopoietic stem cells (HSCs) by lineage depletion and purification based on high aldehyde dehydrogenase activity (ALDH(hi)Lin- cells). Here, we further dissected the ALDH(hi)-Lin- population by selection for CD133, a surface molecule expressed on progenitors from hematopoietic, endothelial, and neural lineages. ALDH(hi)CD133+Lin- cells were primarily CD34+, but also included CD34-CD38-CD133+ cells, a phenotype previously associated with repopulating function. Both ALDH(hi)CD133-Lin- and ALDH(hi)CD133+Lin- cells demonstrated distinct clonogenic progenitor function in vitro, whereas only the ALDH(hi)CD133+Lin- population seeded the murine bone marrow 48 hours after transplantation. Significant human cell repopulation was observed only in NOD/SCID and NOD/SCID beta2M-null mice that received transplants of ALDH(hi)CD133+Lin- cells. Limiting dilution analysis demonstrated a 10-fold increase in the frequency of NOD/SCID repopulating cells compared with CD133+Lin- cells, suggesting that high ALDH activity further purified cells with repopulating function. Transplanted ALDH(hi)CD133+Lin- cells also maintained primitive hematopoietic phenotypes (CD34+CD38-) and demonstrated enhanced repopulating function in recipients of serial, secondary transplants. Cell selection based on ALDH activity and CD133 expression provides a novel purification of HSCs with long-term repopulating function and may be considered an alternative to CD34 cell selection for stem cell therapies. PMID- 16269621 TI - Pathogenic anti-beta2-glycoprotein I antibodies recognize domain I of beta2 glycoprotein I only after a conformational change. AB - Recently, we published the existence of 2 populations of anti-beta2-glycoprotein I (beta2-GPI) IgG antibodies. Type A antibodies recognize epitope G40-R43 in domain I of beta2-GPI and are strongly associated with thrombosis. Type B antibodies recognize other parts of beta2-GPI and are not associated with thrombosis. In this study we demonstrate that type A antibodies only recognize plasma-purified beta2-GPI when coated onto a negatively charged surface and not when coated onto a neutrally charged surface. The affinity of type B antibodies toward plasma-purified beta2-GPI was independent of the charge of the surface to which beta2-GPI was coated. Type A antibodies did not recognize plasma-purified beta2-GPI in solution, whereas they did recognize recombinant beta2-GPI both in solution and coated onto a neutrally charged plate. When the carbohydrate chains were removed from plasma-purified beta2-GPI, we found that type A antibodies did recognize the protein in solution. This supports the hypothesis that the difference in recognition of plasma-purified and recombinant beta2-GPI is caused by the difference in glycosylation and that epitope G40-R43 of plasma-purified beta2-GPI is covered by a carbohydrate chain. Type A anti-beta2-GPI antibodies can only recognize this epitope when this carbohydrate chain is displaced as a result of a conformational change. This finding has major implications both for the detection of pathogenic anti-beta2-GPI antibodies and the comprehension of the pathophysiology of the antiphospholipid syndrome. PMID- 16269622 TI - A distinct and unique transcriptional program expressed by tumor-associated macrophages (defective NF-kappaB and enhanced IRF-3/STAT1 activation). AB - To identify the molecular basis underlying the functions of tumor-associated macrophages (TAMs), we characterized the gene expression profile of TAMs isolated from a murine fibrosarcoma in comparison with peritoneal macrophages (PECs) and myeloid suppressor cells (MSCs), using a cDNA microarray technology. Among the differentially expressed genes, 15 genes relevant to inflammation and immunity were validated by real-time polymerase chain reaction (PCR) and protein production. Resting TAMs showed a characteristic gene expression pattern with higher expression of genes coding for the immunosuppressive cytokine IL-10, phagocytosis-related receptors/molecules (Msr2 and C1q), and inflammatory chemokines (CCL2 and CCL5) as expected, as well as, unexpectedly, IFN-inducible chemokines (CXCL9, CXCL10, CXCL16). Immunohistology confirmed and extended the in vitro analysis by showing that TAMs express M2-associated molecules (eg, IL-10 and MGL1), as well as CCL2, CCL5, CXCL9, CXCL10, and CXCL16, but no appreciable NOS2. Lipopolysaccharide (LPS)-mediated activation of TAMs resulted in defective expression of several proinflammatory cytokines (eg, IL-1beta, IL-6, TNF-alpha) and chemokines (eg, CCL3), as opposed to a strong up-regulation of immunosuppressive cytokines (IL-10, TGFbeta) and IFN-inducible chemokines (CCL5, CXCL9, CXCL10, CXCL16). Thus, profiling of TAMs from a murine sarcoma revealed unexpected expression of IFN-inducible chemokines, associated with an M2 phenotype (IL-10high, IL-12low), and divergent regulation of the NF-kappaB versus the IRF-3/STAT1 pathway. PMID- 16269623 TI - HLAMatchmaker-driven analysis of responses to HLA-typed platelet transfusions in alloimmunized thrombocytopenic patients. AB - This study describes a novel application of HLAMatchmaker to determine platelet compatibility in 16 alloimmunized patients with aplastic anemia refractory to random donor platelet transfusions. HLAMatchmaker is a software algorithm that predicts HLA compatibility by identifying immunogenic epitopes represented by amino acid triplets in antibody-accessible regions of human leukocyte antigen (HLA) molecules and determines the number of triplet mismatches (TMMs) and highly immunogenic triplet mismatches (HIMMs). Corrected count increments (CCIs) and molecular HLA typing were available for 523 transfusions. Conventional compatibility assessment based on cross-reactive group (CREG) determination was not predictive of transfusion outcome. Low HIMMs and TMMs numbers were associated with a higher likelihood of satisfactory (CCIs > or = 8) compared with unsatisfactory (CCIs < 8) outcomes (median HIMMs = 4 vs 6, p2 value < .001; median TMMs = 11 vs 13, p2 value < .001). Although receiver operator characteristic curves revealed that HIMMs or TMMs number are not powerful predictors of individual transfusion outcome, a threshold of at least 3 HIMMs or at least 9 TMMs appeared to be associated with successful transfusions. Triplet matched transfusions were successful, regardless of CREG matching. Our data validate HLAMatchmaker for platelet transfusions and demonstrate its potential to refine and expand donor selection for HLA-alloimmunized patients. PMID- 16269624 TI - The development of a high-density canine microarray. AB - DNA microarrays can give global transcriptional views of cellular responses to disease, development, nutrition, and other biological states. They can be used to elucidate biological networks, develop diagnostics, and identify genetic targets and molecular mechanisms. The technology is widely used and can be a valuable complement to more "disease-centric" focused arrays. For these reasons, Nestle designed a custom canine Affymetrix microarray representing transcripts from multiple tissues for use in areas where a more focused microarray had not already been developed. Sufficient numbers of sequences representing messenger RNAs (mRNAs) or expressed sequence tags (ESTs) is integral for the design of a global microarray chip. This chip was designed using public domain sequences (GenBank) and sequences from a proprietary canine EST database. In order to enrich the chip with annotated transcripts, both of these sequence sets were BLASTed against the nonredundant protein database. The sequences on the microarray were isolated from more than 48 different tissues. The final compliment of sequences had sequences unique to GenBank (3160), unique to the proprietary EST database (17,620), and present in both sources (1996). In comparison with human sequences (RefSeq), 74% of the canine sequences matched a human sequence. PMID- 16269625 TI - Advancing our understanding of the ethics of schizophrenia research: the contribution of conceptual analyses and empirical evidence. PMID- 16269626 TI - alpha2,6-Sialylation promotes binding of placental protein 14 via its Ca2+ dependent lectin activity: insights into differential effects on CD45RO and CD45RA T cells. AB - Placental protein 14 (PP14; glycodelin) is a pregnancy-associated immunoregulatory protein that is known to inhibit T cells via T-cell receptor desensitization. The recent demonstration of PP14 as lectin has provided insight into how it may mediate its CD45 glycoprotein-dependent T-cell inhibition. In this study, we have investigated PP14's lectin-binding properties in detail. Significantly, PP14 reacts with N-acetyllactosamine (LacNAc) as was also found for members of the galectin family, such as the potent immunoregulatory protein, galectin-1. However, in contrast to galectin-1, PP14's binding is significantly enhanced by alpha2,6-sialylation and also by the presence of cations. This was demonstrated by preferential binding to fetuin as compared with its desialylated variant asialofetuin (ASF) and by using free alpha2,6- versus alpha2,3-sialylated forms of LacNAc in competitive inhibition and direct solid-phase binding assays. Interestingly, from immunological point of view, PP14 also binds differentially to CD45 isoforms known to differ in their degree of sialylation. PP14 preferentially inhibits CD45RA+, as compared with CD45RO+ T cells, and preferentially co-capped this variant CD45 on the T-cell surface. Finally, we demonstrate that PP14 promotes CD45 dimerization and clustering, a phenomenon that may regulate CD45 activity. PMID- 16269628 TI - Use of phenytoin and other anticonvulsant prophylaxis in patients with aneurysmal subarachnoid hemorrhage. PMID- 16269629 TI - Hemodynamic evaluation of embolic trajectory in an arterial bifurcation: an in vitro experimental model. AB - BACKGROUND AND PURPOSE: Despite the importance of embolism as a major cause of brain infarction, little is known about the hemodynamic factors governing the path large emboli tend to follow. Our aim was to test in vitro, whether hemodynamic parameters other than flow ratios between bifurcation branches may affect the distribution of embolic particles in a Y-shaped bifurcation model, used as an analogue to an arterial bifurcation. METHODS: In vitro experiments were conducted using suspensions of sphere-shaped particles (0.6, 1.6, and 3.2 mm) in water-glycerin mixture, using steady and pulsatile laminar flow regimes in a Y-shaped bifurcation model (identical branching angles [theta1=theta2=45 degrees] with one daughter branch diameter wider than the other [D1=6 mm, D2=4 mm]; average Reynolds number 500). RESULTS: Experiments using naturally buoyant particles under steady flow conditions and four outlet-flow ratios revealed that small (0.6 mm) and mid-sized (1.6 mm) particles entered into either the narrower or wider bifurcation daughter branch nonpreferentially, proportionally to the flow ratios. Large particles (3.2 mm), however, preferentially entered the wider daughter branch. Moreover, as the flow ratio increases this phenomenon was augmented. Further experiments revealed that the preference of the wider daughter branch for high particle-to-branch diameter-ratios further increases under pulsatile flow and by the density ratio between particles and fluid. CONCLUSIONS: Particles' distribution in a bifurcation is affected, beyond its outlets-flow ratios, by the particle-to-branch diameter-ratio. The tendency of large particles to preferentially enter the wider bifurcation branch, beyond the flow ratio, is augmented under pulsatile flow conditions and is affected by particle-to-fluid density-ratio. These findings may have important implications for understanding the hemodynamic mechanisms underlying the trajectory of large emboli. PMID- 16269630 TI - Dysphagia after stroke: incidence, diagnosis, and pulmonary complications. AB - OBJECTIVE: To determine the incidence of dysphagia and associated pulmonary compromise in stroke patients through a systematic review of the published literature. METHODS: Databases were searched (1966 through May 2005) using terms "cerebrovascular disorders," "deglutition disorders," and limited to "humans" for original articles addressing the frequency of dysphagia or pneumonia. Data sources included Medline, Embase, Pascal, relevant Internet addresses, and extensive hand searching of bibliographies of identified articles. Selected articles were reviewed for quality, diagnostic methods, and patient characteristics. Comparisons were made of reported dysphagia and pneumonia frequencies. The relative risks (RRs) of developing pneumonia were calculated in patients with dysphagia and confirmed aspiration. RESULTS: Of the 277 sources identified, 104 were original, peer-reviewed articles that focused on adult stroke patients with dysphagia. Of these, 24 articles met inclusion criteria and were evaluated. The reported incidence of dysphagia was lowest using cursory screening techniques (37% to 45%), higher using clinical testing (51% to 55%), and highest using instrumental testing (64% to 78%). Dysphagia tends to be lower after hemispheric stroke and remains prominent in the rehabilitation brain stem stroke. There is increased risk for pneumonia in patients with dysphagia (RR, 3.17; 95% CI, 2.07, 4.87) and an even greater risk in patients with aspiration (RR, 11.56; 95% CI, 3.36, 39.77). CONCLUSIONS: The high incidence for dysphagia and pneumonia is a consistent finding with stroke patients. The pneumonia risk is greatest in stroke patients with aspiration. These findings will be valuable in the design of future dysphagia research. PMID- 16269631 TI - Sex hormones and carotid atherosclerosis in the metabolic syndrome. PMID- 16269632 TI - Role of endothelial NO synthase phosphorylation in cerebrovascular protective effect of recombinant erythropoietin during subarachnoid hemorrhage-induced cerebral vasospasm. AB - BACKGROUND AND PURPOSE: In the present study, the effect of subarachnoid hemorrhage (SAH) on the phosphorylation of endothelial NO synthase (eNOS) and the ability of recombinant erythropoietin (Epo) to augment this vasodilator mechanism in the spastic arteries were studied. METHODS: Recombinant adenoviral vectors (10(9) plaque-forming units per animal) encoding genes for human Epo (AdEpo), and beta-galactosidase were injected immediately after injection of autologous arterial blood into the cisterna magna (day 0) of rabbits. Cerebral angiography was performed on day 0 and day 2, and basilar arteries were harvested for Western blots, measurement of cGMP levels, and analysis of vasomotor functions. RESULTS: Injection of autologous arterial blood into cisterna magna resulted in significant vasospasm of the basilar arteries. Despite the narrowing of arterial diameter and reduced expression of eNOS, expressions of phosphorylated protein kinase B (Akt) and phosphorylated eNOS were significantly increased in spastic arteries. Gene transfer of AdEpo reversed the vasospasm. AdEpo-transduced basilar arteries demonstrated significant augmentation of the endothelium-dependent relaxations to acetylcholine, whereas the relaxations to an NO donor, 2-(N,N diethylamino)diazenolate-2-oxide sodium salt, were not affected. Transduction with AdEpo further increased the expression of phosphorylated Akt and eNOS and elevated basal levels of cGMP in the spastic arteries. CONCLUSIONS: Phosphorylation of eNOS appears to be an adaptive mechanism activated during development of vasospasm. The vascular protective effect of Epo against cerebral vasospasm induced by SAH may be mediated in part by phosphorylation of Akt/eNOS. PMID- 16269633 TI - A model of mini-embolic stroke offers measurements of the neurovascular unit response in the living mouse. AB - BACKGROUND AND PURPOSE: To measure cerebral vascular and neuronal responses after stroke in the living mouse, we generated a mouse model of embolic stroke localized to the parietal cortex. METHODS: Male C57/6J or male transgenic mice (2 to 3 months old) expressing yellow fluorescent protein (YFP) were used in the present study. A single fibrin-rich clot (8 mm in length) was injected into a branch of the right middle cerebral artery (MCA). MRI measurements were performed to measure ischemic lesion. Using confocal and 2-photon microscopy, changes in the embolus, dendrites, and dendritic spines were measured in the living mouse. RESULTS: Eight of 11 mice (73%) had the embolus localized to a branch of the right MCA in the parietal cortex. Expansion of the embolus within the artery was observed 24 hours after stroke. The presence of ischemic lesion in the parietal cortex was verified by MRI measurements, and histopathological analysis revealed that these mice (n=8) had a cortical infarct volume of 4.9+/-3.6% of the contralateral hemisphere. In the living mouse, substantial loss of YFP-labeled axonal and dendritic structures as well as the formation of abnormal dendritic bulbs were detected in the ischemic boundary regions 24 hours after stroke compared with that 1 hour after stroke. CONCLUSIONS: This model offers a novel approach to study the neurovascular unit in cerebral cortex after stroke in the living mouse. PMID- 16269634 TI - Polyamine oxidase and acrolein as novel biochemical markers for diagnosis of cerebral stroke. AB - BACKGROUND AND PURPOSE: We found previously that plasma levels of acrolein (CH2=CHCHO) and spermine oxidase (SMO) were well correlated with the degree of severity of chronic renal failure. The aim of this study was to test whether the levels of these 2 markers and of acetylpolyamine oxidase (AcPAO) were increased in the plasma of stroke patients. METHODS: The activity of AcPAO and SMO and the level of protein-conjugated acrolein in plasma of the stroke patients and normal subjects were measured by high-performance liquid chromatography and ELISA, respectively. Focal infarcts were estimated by MRI or computed tomography (CT). RESULTS: The levels of AcPAO, SMO, and acrolein were significantly increased in the plasma of stroke patients. The size of stroke was nearly parallel with the multiplied value of acrolein and total polyamine oxidase (AcPAO plus SMO). After the onset of stroke, an increase in AcPAO first occurred, followed by increased levels of SMO and finally acrolein. In 1 case, an increase in AcPAO and SMO preceded focal damage as detected by MRI or CT. Furthermore, stroke was confirmed by MRI in a number of mildly symptomatic patients (11 cases) who had increased levels of total polyamine oxidase and acrolein. Among apparently normal subjects (8 cases) who had high values of acroleinxtotal polyamine oxidase, stroke was found in 4 cases by MRI. CONCLUSIONS: The results indicate that increased levels of AcPAO, SMO, and acrolein are good markers of stroke. PMID- 16269635 TI - Constrictor and dilator effects of angiotensin II on cerebral arterioles. AB - BACKGROUND AND PURPOSE: In light of the equivocal data on the cerebral vasoconstrictor and vasodilator actions of angiotensin II (Ang II) and the potential clinical importance of this, we investigated the effects of Ang II on rat pial arterioles. METHODS: We determined the effect of Ang I (3.10(-6) mol/L) in the absence and presence of the converting enzyme inhibitor, captopril (10(-5) mol/L) in cerebral arterioles of male Wistar rats (open-skull preparation), and those of Ang II (3.10(-12) to 3.10(-6) mol/L) in the absence and presence of the Ang II receptor (AT1) antagonist, telmisartan (10(-5) mol/L) or the AT2 antagonist, PD123319 (10(-5) mol/L). We examined the effect of PD123319 (10(-5) mol/L) and the Ca2+-activated K+ (BKCa) channel blocker, tetraethylammonium (10( 4) mol/L) on the Ang II responses in the presence of telmisartan (10(-5) mol/L). RESULTS: Ang II-induced dose-dependent constriction with a maximum decrease of 20.1+/-1.0% at 10(-6) mol/L. Captopril significantly decreased Ang I-induced vasoconstriction (-4.0+/-0.9 versus -21.3+/-2.5%; n=4). Telmisartan reversed Ang II-induced vasoconstriction (9.5+/-2.5 versus -20.1+/-1% at 10(-6) mol/L; n=5). PD123319 significantly increased Ang II-induced vasoconstriction (-12.9+/-0.8 versus -10.2+/-0.4% at 10(-6) mol/L; n=5). PD123319 abolished (-2.6+/-0.7 versus 9.3+/-1.1% at 10(-6) mol/L; n=5) whereas tetraethylammonium reversed (-12.1+/-1.6 versus 9.9+/-1.0% at 10(-6) mol/L; n=4) Ang II-induced vasodilatation in the presence of telmisartan. CONCLUSIONS: Angiotensin is converted locally into Ang II; the overall effect of Ang II is vasoconstrictor following stimulation of the AT1 receptor, but a vasodilator response can be evoked following stimulation of the AT2 receptor and activation of BKCa. PMID- 16269636 TI - Metalloporphyrin-based superoxide dismutase mimic attenuates the nuclear translocation of apoptosis-inducing factor and the subsequent DNA fragmentation after permanent focal cerebral ischemia in mice. AB - BACKGROUND AND PURPOSE: Recently, apoptosis- inducing factor (AIF), a mitochondrial proapoptotic protein, and its nuclear translocation have been reported in caspase-independent neuronal apoptosis. In this study, we investigated the contribution of reactive oxygen species (ROS) to the nuclear translocation of AIF and the subsequent DNA fragmentation after permanent focal cerebral ischemia (pFCI) using manganese tetrakis (4-benzoic acid) porphyrin (MnTBAP), which mimics mitochondrial superoxide dismutase. METHOD: Adult male ICR mice were subjected to pFCI by intraluminal suture blockade of the middle cerebral artery. Immunohistochemistry and Western blot analysis were performed. Large-scale DNA fragmentation was evaluated by pulse field gel electrophoresis, and apoptotic cell death was quantified. MnTBAP was injected into the ventricle to determine whether the removal of ROS contributes to AIF translocation and the subsequent DNA fragmentation. RESULTS: Western blot analysis showed that the nuclear translocation of AIF occurred as early as 2 hours after pFCI. AIF translocation was not blocked by a pan-caspase inhibitor. MnTBAP-treated mice had attenuated AIF translocation and blocked large-scale DNA fragmentation. Caspase-3 activity was similarly inhibited between the pan-caspase inhibitor- and MnTBAP treated mice, but the amount of apoptosis-associated DNA fragmentation in the MnTBAP-treated mice was less than in the pan-caspase inhibitor-treated mice (P<0.001). CONCLUSIONS: These results suggest that the MnTBAP, a mitochondrial O2 scavenger, may attenuate the caspase-independent nuclear translocation of AIF after pFCI and subsequent apoptosis-associated DNA fragmentation. PMID- 16269637 TI - Lesion patterns and stroke mechanism in atherosclerotic middle cerebral artery disease: early diffusion-weighted imaging study. AB - BACKGROUND AND PURPOSE: Patterns and mechanisms of stroke in patients with atherosclerotic middle cerebral artery (MCA) disease remain unclear. We sought to identify lesion patterns and stroke mechanisms associated with MCA disease using early diffusion-weighted imaging (DWI). METHODS: We reviewed 185 acute ischemic stroke patients who had (1) symptomatic lesions located in the unilateral MCA territory on DWI performed within 48 hours of symptom onset, and (2) either corresponding MCA disease, internal carotid artery disease, or cardioembolism. Acute DWI lesion patterns were classified as (1) single (small perforator <2 cm; large perforator > or =2 cm; pial; large territorial; border-zone) and (2) multiple. RESULTS: MCA disease was diagnosed in 63 patients, 32 (50.8%) of whom showed multiple lesions. Concomitant perforator and pial infarcts (14/63, P<0.001), concomitant perforator, pial, and border-zone infarcts (9/63, P<0.001), and single small perforator infarcts (12/63, P=0.001) were identified more often in patients with MCA disease than in those with cardioembolism or internal carotid artery disease. Small perforator infarcts were more common in patients with milder stenosis than with severe stenosis or occlusion of MCA (P<0.001). Whether they occurred singly or in addition to other lesions, pial infarcts were identified more often in patients with severe stenosis or occlusion of MCA (P=0.001). CONCLUSIONS: Perforating artery infarcts, whether single or occurring in addition to pial or border-zone infarcts, are lesion patterns specific for MCA disease. This suggests that local branch occlusion and coexisting distal embolization may be a common stroke mechanism in patients with MCA disease. PMID- 16269638 TI - Facilitation of sensory and motor recovery by thermal intervention for the hemiplegic upper limb in acute stroke patients: a single-blind randomized clinical trial. AB - BACKGROUND AND PURPOSE: Thermal stimulation (TS) is commonly used in orthopedic rehabilitation, but the role of TS in the facilitation of sensorimotor recovery in hemiplegic patients remains unknown. This study addressed the issue of TS intervention in the facilitation of functional outcomes. METHODS: Forty-six stroke survivors were randomly assigned to standard rehabilitation treatment and standard treatment plus TS (30 minutes daily for 6 weeks). Twenty-nine patients completed the experiment. Six measures, including Brunnstrom stage, modified motor assessment scale, grasping strength, angles of wrist extension and flexion, sensation by monofilament, and muscle tone by modified Ashworth scale, were performed weekly to evaluate sensory and motor functional outcomes. RESULTS: The performance of Brunnstrom stage and wrist extension and sensation were improved significantly after TS intervention. Recovery rates of 6 measures after TS were significantly higher than those of the control, except for grasping. Similar muscle tones were found in both groups. CONCLUSIONS: TS on the paretic hand significantly enhances the recovery of several aspects of sensory and motor functions in hemiplegic stroke patients. PMID- 16269639 TI - Conversion of ischemic brain tissue into infarction increases with age. AB - BACKGROUND AND PURPOSE: Brain regions normal on diffusion-weighted imaging (DWI) but abnormal on mean transit time (MTT) maps represent tissue at risk of infarction, yet the fate of these regions is quite variable. The imperfect correlation between tissue outcome and initial imaging parameters suggests that each patient's brain may have different susceptibility to ischemic stress. We hypothesize that age is a marker for tissue susceptibility to ischemia and thus plays a role in determining tissue outcome in human stroke. METHODS: Sixty patients with acute ischemic stroke and a region of DWI/MTT mismatch that was >20% of the DWI volume were included. All patients were scanned twice, within 12 hours of symptom onset and on day 5 or later. The percentage mismatch lost (PML) was calculated as percentage of initial DWI/MTT mismatch volume that was infarcted on the follow-up MRI. The statistical analysis explored relationships among the covariates age, Trial of Org 10172 in Acute Stroke Treatment (TOAST) subtypes, time-to-MRI, and initial DWI, MTT volume, mean arterial blood pressure and blood glucose level at admission, and previous history of hypertension and diabetes mellitus. RESULTS: Univariate comparisons showed that age (P=0.003), hypertension (P=0.009), and diabetes mellitus (P=0.0002) were significantly associated with PML. Regression analyses showed age to be a significant covariate (P=0.02). The regression model predicted a change in PML of approximately 0.65% per year. The adjusted proportion of variance (R2) in PML that could be explained by age alone was 14%. CONCLUSIONS: Age-dependent increase in conversion of ischemic tissue into infarction suggests that age is a biological marker for the variability in tissue outcome in acute human stroke. PMID- 16269640 TI - Is white matter involved in patients entered into typical trials of neuroprotection? AB - BACKGROUND AND PURPOSE: One of the reasons for the failure of trials of neuroprotection in stroke may be the lack of white matter (WM) protection. However, whether patients entered into typical neuroprotection trials have WM involved in the ischemic process is unknown. We studied patients who were enrolled in neuroprotection trials at our center and used a neuroimaging coregistration approach to determine whether final infarcts involved WM and, if so, in what proportion. We also aimed to provide the first in vivo volume distribution of gray matter (GM) and WM in normal stroke-aged brains. METHODS: Patients enrolled in trials of neuroprotection had late computed tomography or magnetic resonance scans coregistered in standard stereotaxic coordinate space after segmentation of symptomatic cerebral infarcts. These were then superimposed on a probabilistic map of GM and WM, which was developed from age-matched normal controls in whom GM and WM volumes were assessed. RESULTS: Forty-two patients (mean age, 73.7+/-10.5 years) were studied from 6 trials of neuroprotection. WM formed 41.7% of the brain volume in 37 control subjects (mean age, 73.5+/-8.4 years). In the segmented infarcts, WM comprised a median of 49% (interquartile range, 36.5 to 77.9) of the infarct volume. Ninety-five percent of infarcts had some involvement of WM tracts. CONCLUSIONS: WM occupies approximately 42% by volume of the normal stroke-aged brain. Patients entered into typical trials of neuroprotection may have significant WM volumes involved in the ischemic process, thus providing a rationale for the development of neuroprotectants for this compartment. PMID- 16269641 TI - Fibrinogen is associated with an increased risk of Alzheimer disease and vascular dementia. AB - BACKGROUND AND PURPOSE: Vascular and inflammatory factors may play an important role in the pathogenesis of dementia. Studies reported an association between plasma levels of inflammation markers and the risk of dementia. Both fibrinogen and C-reactive protein are considered inflammatory markers. Fibrinogen also has important hemostatic properties. We investigated the association of fibrinogen and C-reactive protein with dementia. METHODS: The study was based on the prospective population-based Rotterdam Study. Fibrinogen was measured in a random sample of 2835 persons. High-sensitivity C-reactive protein was measured in the total cohort of 6713 persons. We identified 395 incident dementia cases during follow-up (mean, 5.7 years). We estimated the associations of fibrinogen and C reactive protein with dementia using Cox proportional hazard models. RESULTS: Persons with higher levels of fibrinogen had an increased risk of dementia. The hazard ratio for dementia per SD increase of fibrinogen was 1.26 (95% CI, 1.11 to 1.44), adjusted for age and gender, and 1.30 (95% CI, 1.13 to 1.50) after additional adjustment for cardiovascular factors and stroke. For Alzheimer disease, the adjusted hazard ratio was 1.25 (95% CI, 1.04 to 1.49), and for vascular dementia it was 1.76 (95% CI, 1.34 to 2.30). High levels of C-reactive protein were not associated with an increased risk of dementia. CONCLUSIONS: High fibrinogen levels were associated with an increased risk of both Alzheimer disease and vascular dementia, but levels of C-reactive protein were not. This suggests that the increased risk of dementia associated with fibrinogen is because of the hemostatic rather than the inflammatory properties of fibrinogen. PMID- 16269642 TI - From trials to "real life": necessity of efficacy. PMID- 16269643 TI - Diffusion histograms in CADASIL. PMID- 16269644 TI - Diffusion magnetic resonance histograms as a surrogate marker and predictor of disease progression in CADASIL: a two-year follow-up study. AB - BACKGROUND AND PURPOSE: Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is a cerebral small vessel disease caused by mutations in the NOTCH3 gene. MRI is sensitive in detecting preclinical involvement and changes over time. However, little is known about correlations between MRI metrics and clinical measures on a longitudinal scale. In this study, we assessed the role of quantitative MRI (T2-lesion volume and diffusion tensor imaging [DTI]-derived metrics) in monitoring and predicting disease progression. METHODS: Sixty-two CADASIL subjects were followed prospectively over a period of 26.3+/-1.2 months. Dual-echo scans, DTI scans, and clinical scales were obtained at baseline and at follow-up. T2-lesion volumes were determined quantitatively, and histograms of mean diffusivity (MD) were produced. RESULTS: At follow-up, T2 lesion volumes and MD histogram metrics had changed significantly (all P<0.01). Lesion volumes and average MD correlated with clinical scores at baseline. Changes of average MD correlated with changes of the Rankin score, the National Institutes of Health Stroke Scale score, and the structured interview for the diagnosis of Alzheimer dementia and multiinfarct dementia score (all P<0.01). On multivariate analysis, average MD and systolic blood pressure at baseline were predictors of changes of average MD during follow-up. Moreover, average MD was the main predictor of clinical progression. Sample size estimates showed that the number of individuals required to detect a treatment effect in an interventional trial may be reduced when using MD histograms as an end point. CONCLUSIONS: This study establishes correlations between changes of DTI histogram metrics and clinical measures over time. DTI histograms may be used as an adjunct outcome measure in future therapeutic trials. Moreover, DTI histogram metrics predict disease progression in CADASIL. PMID- 16269645 TI - Apparent diffusion coefficient thresholds do not predict the response to acute stroke thrombolysis. AB - BACKGROUND AND PURPOSE: Apparent diffusion coefficient (ADC) thresholds for tissue infarction have been identified in acute stroke. IV tissue plasminogen activator (tPA) is associated with tissue salvage. We hypothesized that tPA would lower the ADC threshold for infarction. METHODS: ADC and mean transit time (MTT) maps were generated for 26 patients imaged within 6 hours of stroke onset (12 tPA and 14 conservatively managed controls). MTT maps and day-90 T2-weighted images were coregistered to ADC maps. Relative ADC (rADC) values were calculated for initial diffusion-weighted imaging (DWI) lesions, infarct growth regions (final infarct volume-the acute DWI lesion volume), and hypoperfused salvaged regions (HS; MTT map abnormality-the final infarct volume). When relevant, the DWI lesion was subdivided into DWI reversal and DWI infarct regions. RESULTS: Mean DWI lesion rADC was 0.79 in tPA and 0.74 in untreated patients (P=0.097). Mean rADC in HS and infarct growth regions were similar in tPA patients (0.950 and 0.946) and untreated patients (0.957, P=0.76; 0.970, P=0.08, respectively). The rADC in HS tissue was directly correlated with the time to treatment with tPA (r=0.685; P=0.029). DWI reversal was seen in 67% of tPA-treated patients and in 36% of those conservatively managed (Fisher exact test; P=0.238). In the 13 patients with DWI reversal, the mean rADC in these regions (0.81+/-0.07) was significantly higher than in the acute DWI region that infarcted (0.74+/-0.07; P=0.02), although no absolute thresholds could be identified. CONCLUSIONS: The peri-DWI lesion region contains tissue with intermediate ADC values. The fate of this tissue is variable and cannot be predicted based on the ADC alone. DWI expansion occurs in bioenergetically normal tissue, and this is attenuated by tPA in a time dependent fashion. PMID- 16269646 TI - Potential applicability of recombinant factor VIIa for intracerebral hemorrhage. AB - BACKGROUND AND PURPOSE: To date, there are no proven, effective treatments for intracerebral hemorrhage (ICH) beyond supportive medical care. A recent randomized, blinded, placebo-controlled trial of recombinant factor VIIa (rFVIIa) administered intravenously within 4 hours of ICH onset reported a reduction in morbidity and mortality compared with placebo. We sought to determine the potential applicability of rFVIIa in a large, population-based cohort of ICH patients. METHODS: All of the patients age > or =18 years hospitalized with nontraumatic ICH in the Greater Cincinnati region were identified from May 1998 to July 2001 and August 2002 to April 2003. Patient demographics were compared with the inclusion and exclusion criteria from the rFVIIa trial to determine eligibility for treatment and reasons for exclusion. Mortality in the eligible patient group was compared with the placebo group in the rFVIIa trial. RESULTS: Over 4 calendar years, 1018 ICH patients were identified; of these, 133 (13.1%) had no exclusions and presented within the prescribed time window. An additional 45 patients (4.4%) may have been eligible but had uncertain onset or computed tomography scan times. The most common reasons for exclusion (not mutually exclusive) were late presentation (n=398), vaso-occlusive disease (n=369), deep coma (n=219), and prolonged international normalized ratio or partial thromboplastin time (n=200). Mortality at 90 days among potentially eligible patients was the same as for the placebo group in the rFVIIa trial (29% versus 29%; P=0.99). CONCLUSIONS: In this large, population-based ICH cohort, 13.1% to 17.5% of patients would have qualified for treatment with rFVIIa by trial criteria. PMID- 16269647 TI - Motor strokes: the lesion location determines motor excitability changes. AB - BACKGROUND AND PURPOSE: The purpose of this research was to investigate the impact of lesion location on motor excitability and motor performance. METHODS: We studied patients with pure motor strokes in 4 different brain areas: motor cortex lesions (n=7), striatocapsular lesions (n=13), lacunar lesions of the internal capsule (n=13), and paramedian pontine lesions (n=10). Motor performance tests included the 9-hole-peg test and grip strength recordings. Motor excitability was determined by transcranial magnetic stimulation. Motor thresholds, stimulus-response curves, silent periods, motor cortical inhibition, and facilitation were investigated. RESULTS: The 4 groups were clinically similar but showed major differences in motor excitability. Only motor cortex lesions had a loss of intracortical inhibition in the affected hemisphere. In the internal capsule lesion group and the pontine lesion group, stimulus-response curves were depressed on the affected side. All of the subcortical lesions showed a prolongation of the silent period in the paretic side. Motor thresholds were predominantly elevated in the lesioned hemisphere of patients with internal capsule or pontine lesions. Motor performance was correlated with silent period duration in internal capsule lesions and with motor thresholds in internal capsule and pontine lesions. CONCLUSIONS: Motor cortex lesions exhibited deficient inhibitory properties. In contrast, subcortical lesions displayed an enhancement of inhibition. Internal capsule and pontine lesions affecting the corticospinal tract on different levels particularly impaired neuronal recruitment. Our results suggest that the lesion location determines a specific pattern of motor excitability changes. PMID- 16269648 TI - Intracranial aneurysms treated with endovascular coils: detection of recurrences using unenhanced and contrast-enhanced transcranial color-coded duplex sonography. AB - BACKGROUND AND PURPOSE: Because neck recurrence after endovascular treatment of intracranial aneurysms (IAs) is not uncommon, surveillance to assess long-term stability of occlusion is clearly important. This study evaluated unenhanced and contrast-enhanced transcranial color-coded duplex sonography (TCCS) in detecting refilling of IAs treated with detachable coils. METHODS: Patients with coiled IAs were imaged before and after contrast enhancement. The results were compared with those of a surveillance digital subtraction angiogram (DSA). The operator was blinded to the results of the DSA. Aneurysms were classed as either occluded or with residual flow and quantified as minor, moderate, or extensive. There were 208 studies performed in 4 neurosurgical centers. Of those, 141 studies received ultrasonic contrast enhancement with Levovist, and 68 had an additional enhanced study with SonoVue. RESULTS: We excluded 44 studies. Of the 164 unenhanced studies, TCCS correctly identified 52 of 67 cases defined as completely occluded by DSA (sensitivity 78%; specificity 77%), 13 of 50 aneurysms with minor refilling (sensitivity 26%; specificity 88%), 15 of 27 aneurysms with moderate refilling (sensitivity 56%; specificity 95%), and 9 of 20 aneurysms with extensive refilling (sensitivity 45%; specificity 100%). TCCS correctly identified an additional 10 aneurysms with minor refilling after Levovist enhancement and 3 with SonoVue. Both SonoVue and Levovist enhancement identified an additional 1 aneurysm with moderate refilling and 3 with extensive refilling. CONCLUSIONS: TCCS could be used to selectively monitor IAs, which would reduce the requirement for long-term invasive monitoring. The detection of neck refilling is improved with contrast enhancement. PMID- 16269649 TI - Functional recovery after hemiplegia in patients with neglect: the rehabilitative role of anosognosia. AB - BACKGROUND AND PURPOSE: The aim of this study was to verify whether the presence of anosognosia (A) affects the rehabilitative prognosis of hemiplegic subjects with neglect (N). METHODS: This study was carried out on 30 patients with left hemiplegia: 15 patients had neglect (group N) and 15 had neglect and anosognosia (group N+A). Mean age was 68.2+/-6.3 in group N (9 men and 6 women) and 72.1+/ 6.4 in group N+A (7 men and 8 women). The average interval from onset of stroke to admission for rehabilitation was 23 and 23.6 days, respectively, in group N and in group N+A. Patients were assessed through the Mesulam test, Bisiach test, Wechsler Adult Intelligence Scale, Fugl-Meyer scale, Functional Independence Measure (FIM), and Rankin scale. RESULTS: Before rehabilitation, cognitive FIM scores of patients of group N were significantly higher than those of group N+A (P=0.001), whereas motor FIM scores and total FIM scores did not differ between the 2 groups. After rehabilitation, cognitive FIM scores (P=0.000) and even motor (P=0.009) and total FIM scores (P=0.000) were statistically higher in group N than in group N+A. Effectiveness (P=0.005) and efficiency (P=0.012) in the motor FIM scores of group N were significantly greater than those of group N+A. Disability was lower in group N (P=0.040). CONCLUSIONS: Our study shows that the presence of anosognosia worsens the rehabilitation prognosis in hemiplegic subjects who also have neglect. PMID- 16269650 TI - Detection of thrombus in acute ischemic stroke: value of thin-section noncontrast computed tomography. AB - BACKGROUND AND PURPOSE: Previous studies on the hyperdense middle cerebral artery (MCA) sign were conducted using > or =5-mm thickness noncontrast-computed tomography (NCT). The purpose of this study was to compare thin-section NCT with 5-mm NCT in the detection of thrombus in acute ischemic stroke. METHODS: Enrolled were consecutive 51 patients with acute infarction in the anterior or MCA territory. All patients underwent both 5-mm NCT and either 1.25- or 1-mm thin section helical NCT within 6 hours of symptom onset. Patients were assigned to either the single or multisegmental occlusion group, depending on the thrombus extent on thin-section NCT. Thin-section NCT and 5-mm NCT were compared in the detection of thrombi. RESULTS: Thrombi were identified in 45 patients (88%) on thin-section NCT and 16 on 5-mm NCT (31%; P<0.001). No occlusion was seen in 6 patients. Both sensitivity and specificity of thin-section NCT in detection of thrombus were 100%. CONCLUSIONS: Acute thrombus can be detected with higher sensitivity on thin-section NCT than on 5-mm NCT, and its extent is more accurately determined on thin-section NCT. PMID- 16269651 TI - Increased neointima formation in cysteine-rich protein 2-deficient mice in response to vascular injury. AB - In response to arterial injury, medial vascular smooth muscle cells (VSMCs) proliferate and migrate into the intima, contributing to the development of occlusive vascular disease. The LIM protein cysteine-rich protein (CRP) 2 associates with the actin cytoskeleton and may maintain the cytoarchitecture. CRP2 also interacts with transcription factors in the nucleus to mediate SMC gene expression. To test the hypothesis that CRP2 may be an important regulator of vascular development or function we generated Csrp2 (gene symbol of the mouse CRP2 gene)-deficient (Csrp2(-/-)) mice by targeted mutation. Csrp2(-/-) mice did not have any gross vascular defects or altered expression levels of SM alpha actin, SM22alpha, or calponin. Following femoral artery injury, CRP2 expression persisted in the vessel wall at 4 days and then decreased by 14 days. Intimal thickening was enhanced 3.4-fold in Csrp2(-/-) compared with wild-type (WT) mice 14 days following injury. Cellular proliferation was similar between WT and Csrp2(-/-) VSMC both in vivo and in vitro. Interestingly, Csrp2(-/-) VSMC migrated more rapidly in response to PDGF-BB and had increased Rac1 activation. Our data demonstrate that CRP2 is not required for vascular development. However, an absence of CRP2 enhanced VSMC migration and increased neointima formation following arterial injury. PMID- 16269652 TI - Restoration of cardiac progenitor cells after myocardial infarction by self proliferation and selective homing of bone marrow-derived stem cells. AB - Tissue-specific progenitor cells contribute to local cellular regeneration and maintain organ function. Recently, we have determined that cardiac side population (CSP) cells represent a distinct cardiac progenitor cell population, capable of in vitro differentiation into functional cardiomyocytes. The response of endogenous CSP to myocardial injury, however, and the cellular mechanisms that maintain this cardiac progenitor cell pool in vivo remain unknown. In this report we demonstrate that local progenitor cell proliferation maintains CSP under physiologic conditions, with little contribution from extracardiac stem cell sources. Following myocardial infarction in adult mice, however, CSP cells are acutely depleted, both within the infarct and noninfarct areas. CSP pools are subsequently reconstituted to baseline levels within 7 days after myocardial infarction, through both proliferation of resident CSP cells, as well as through homing of bone marrow-derived stem cells (BMC) to specific areas of myocardial injury and immunophenotypic conversion of BMC to adopt a CSP phenotype. We, therefore, conclude that following myocardial injury, cardiac progenitor cell populations are acutely depleted and are reconstituted to normal levels by both self-proliferation and selective homing of BMC. Understanding and enhancing such processes hold enormous potential for therapeutic myocardial regeneration. PMID- 16269653 TI - Ca2+/calmodulin-dependent protein kinase modulates cardiac ryanodine receptor phosphorylation and sarcoplasmic reticulum Ca2+ leak in heart failure. AB - Abnormal release of Ca from sarcoplasmic reticulum (SR) via the cardiac ryanodine receptor (RyR2) may contribute to contractile dysfunction and arrhythmogenesis in heart failure (HF). We previously demonstrated decreased Ca transient amplitude and SR Ca load associated with increased Na/Ca exchanger expression and enhanced diastolic SR Ca leak in an arrhythmogenic rabbit model of nonischemic HF. Here we assessed expression and phosphorylation status of key Ca handling proteins and measured SR Ca leak in control and HF rabbit myocytes. With HF, expression of RyR2 and FK-506 binding protein 12.6 (FKBP12.6) were reduced, whereas inositol trisphosphate receptor (type 2) and Ca/calmodulin-dependent protein kinase II (CaMKII) expression were increased 50% to 100%. The RyR2 complex included more CaMKII (which was more activated) but less calmodulin, FKBP12.6, and phosphatases 1 and 2A. The RyR2 was more highly phosphorylated by both protein kinase A (PKA) and CaMKII. Total phospholamban phosphorylation was unaltered, although it was reduced at the PKA site and increased at the CaMKII site. SR Ca leak in intact HF myocytes (which is higher than in control) was reduced by inhibition of CaMKII but was unaltered by PKA inhibition. CaMKII inhibition also increased SR Ca content in HF myocytes. Our results suggest that CaMKII-dependent phosphorylation of RyR2 is involved in enhanced SR diastolic Ca leak and reduced SR Ca load in HF, and may thus contribute to arrhythmias and contractile dysfunction in HF. PMID- 16269654 TI - Adiponectin inhibits endothelial synthesis of interleukin-8. AB - Adiponectin is an antiatherogenic adipokine that inhibits inflammation by mechanisms that are not completely understood. We explored the effect of adiponectin on endothelial synthesis of interleukin-8 (IL-8), a pro-inflammatory chemokine that plays a role in atherogenesis. Adiponectin decreased the secretion of IL-8 from human aortic endothelial cells (HAEC) stimulated with tumor necrosis factor-alpha (TNF-alpha). Adiponectin also inhibited IL-8 mRNA expression induced by TNF-alpha. Phosphorylation of IkappaB-alpha was decreased by adiponectin, but phosphorylation of ERK, SAPK/JNK, and p38MAPK were unaffected. Adiponectin increased intra-cellular cAMP levels in HAEC in a dose-dependent manner; PKA activity was also increased. The inhibitory effect of adiponectin on TNF-alpha induced IL-8 synthesis was inhibited by pretreatment with Rp-cAMP, a PKA inhibitor. These observations suggest that adiponectin inhibits IL-8 synthesis through inhibition of a PKA dependent NF-kappaB signaling pathway. We also showed that adiponectin enhances Akt phosphorylation. The inhibitory effect of adiponectin on TNF-alpha-induced IL-8 synthesis was abrogated in part by pretreatment with the PI3 kinase inhibitor LY294002 or by Akt siRNA transfection, suggesting that Akt activation might inhibit IL-8 synthesis induced by TNF-alpha. We conclude that inhibition of NF-kappaB and activation of Akt phosphorylation may mediate adiponectin inhibition of atherosclerosis. PMID- 16269655 TI - cAMP-binding protein Epac induces cardiomyocyte hypertrophy. AB - cAMP is one of the most important second messenger in the heart. The discovery of Epac as a guanine exchange factor (GEF), which is directly activated by cAMP, raises the question of the role of this protein in cardiac cells. Here we show that Epac activation leads to morphological changes and induces expression of cardiac hypertrophic markers. This process is associated with a Ca2+-dependent activation of the small GTPase, Rac. In addition, we found that Epac activates a prohypertrophic signaling pathway, which involves the Ca2+ sensitive phosphatase, calcineurin, and its primary downstream effector, NFAT. Rac is involved in Epac induced NFAT dependent cardiomyocyte hypertrophy. Blockade of either calcineurin or Rac activity blunts the hypertrophic response elicited by Epac indicating these signaling molecules coordinately regulate cardiac gene expression and cellular growth. Our results thus open new insights into the signaling pathways by which cAMP may mediate its biological effects and identify Epac as a new positive regulator of cardiac growth. PMID- 16269656 TI - Soluble vascular endothelial growth factor receptor-1 (sFLT-1) mediates downregulation of FLT-1 and prevents activated neutrophils from women with preeclampsia from additional migration by VEGF. AB - Neutrophil activation and increased migration is associated with preeclampsia and is resolved after delivery. Preeclampsia is an inflammatory disorder where altered levels of vascular endothelial growth factor (VEGF) and the circulating soluble fms-like tyrosine kinase 1 (sFlt-1) have a pathogenic role. VEGF, by binding to FLT-1, induces leukocytic chemotaxis. We studied expression and function of FLT-1 in maternal neutrophils during preeclampsia and normal pregnancies. Analysis of maternal neutrophils showed the relationship between FLT 1 expression and week of gestation. Preeclamptic women express lower FLT-1 and sFLT-1 in neutrophils. In contrast, serum levels of sFLT-1 in patients with preeclampsia are increased and, therefore, inhibit upregulation of FLT-1 in neutrophils by neutralizing VEGF. VEGF-dependent FLT-1 expression is regulated by changing FLT-1-promoter activity. Promoter activity is decreased by sFLT-1. In vitro experiments demonstrated that migration of neutrophils is regulated by VEGF via FLT-1 and excess of sFLT-1. Thus, VEGF-dependent migration of neutrophils is decreased during preeclampsia as a consequence of excess circulating sFlt1. But, they still increase migration by fMLP and, therefore, migration of neutrophils from preeclamptic women is highly activated when compared with the normotensive group. In conclusion, besides being involved in inducing an antiangiogenic state in the serum, excess of sFLT-1 seems to prevent activated neutrophils from women with preeclampsia from additional migration by VEGF. We provide evidence that neutrophils may be involved in the pathophysiology of pregnancy-related hypertensive disorders. PMID- 16269657 TI - FcgammaRIIB mediates C-reactive protein inhibition of endothelial NO synthase. AB - C-reactive protein (CRP) is an acute-phase reactant that is positively correlated with cardiovascular disease risk and endothelial dysfunction. Whether CRP has direct actions on endothelium and the mechanisms underlying such actions are unknown. Here we show in cultured endothelium that CRP prevents endothelial NO synthase (eNOS) activation by diverse agonists, resulting in the promotion of monocyte adhesion. CRP antagonism of eNOS occurs nongenomically and is attributable to blunted eNOS phosphorylation at Ser1179. Okadaic acid or knockdown of PP2A by short-interference RNA reverses CRP antagonism of eNOS, indicating a key role for the phosphatase. Aggregated IgG, the known ligand for Fcgamma receptors, causes parallel okadaic acid-sensitive loss of eNOS function, FcgammaRIIB expression is demonstrable in endothelium, and heterologous expression studies reveal that CRP antagonism of eNOS requires FcgammaRIIB. In FcgammaRIIB(+/+) mice, CRP blunts acetylcholine-induced increases in carotid artery vascular conductance; in contrast, CRP enhances acetylcholine responses in FcgammaRIIB(-/-) mice. Thus FcgammaRIIB mediates CRP inhibition of eNOS via PP2A, providing a mechanistic link between CRP and endothelial dysfunction. PMID- 16269658 TI - Contribution of Kv channels to phenotypic remodeling of human uterine artery smooth muscle cells. AB - Vascular smooth muscle cells (VSMCs) perform diverse functions that can be classified into contractile and synthetic (or proliferating). All of these functions can be fulfilled by the same cell because of its capacity of phenotypic modulation in response to environmental changes. The resting membrane potential is a key determinant for both contractile and proliferating functions. Here, we have explored the expression of voltage-dependent K+ (Kv) channels in contractile (freshly dissociated) and proliferating (cultured) VSMCs obtained from human uterine arteries to establish their contribution to the functional properties of the cells and their possible participation in the phenotypic switch. We have studied the expression pattern (both at the mRNA and at the protein level) of Kvalpha subunits in both preparations as well as their functional contribution to the K+ currents of VSMCs. Our results indicate that phenotypic remodeling associates with a change in the expression and distribution of Kv channels. Whereas Kv currents in contractile VSMCs are mainly performed by Kv1 channels, Kv3.4 is the principal contributor to K+ currents in cultured VSMCs. Furthermore, selective blockade of Kv3.4 channels resulted in a reduced proliferation rate, suggesting a link between Kv channels expression and phenotypic remodeling. PMID- 16269659 TI - TRPV4 forms a novel Ca2+ signaling complex with ryanodine receptors and BKCa channels. AB - Vasodilatory factors produced by the endothelium are critical for the maintenance of normal blood pressure and flow. We hypothesized that endothelial signals are transduced to underlying vascular smooth muscle by vanilloid transient receptor potential (TRPV) channels. TRPV4 message was detected in RNA from cerebral artery smooth muscle cells. In patch-clamp experiments using freshly isolated cerebral myocytes, outwardly rectifying whole-cell currents with properties consistent with those of expressed TRPV4 channels were evoked by the TRPV4 agonist 4alpha phorbol 12,13-didecanoate (4alpha-PDD) (5 micromol/L) and the endothelium-derived arachidonic acid metabolite 11,12 epoxyeicosatrienoic acid (11,12 EET) (300 nmol/L). Using high-speed laser-scanning confocal microscopy, we found that 11,12 EET increased the frequency of unitary Ca2+ release events (Ca2+ sparks) via ryanodine receptors located on the sarcoplasmic reticulum of cerebral artery smooth muscle cells. EET-induced Ca2+ sparks activated nearby sarcolemmal large conductance Ca2+-activated K+ (BKCa) channels, measured as an increase in the frequency of transient K+ currents (referred to as "spontaneous transient outward currents" [STOCs]). 11,12 EET-induced increases in Ca2+ spark and STOC frequency were inhibited by lowering external Ca2+ from 2 mmol/L to 10 micromol/L but not by voltage-dependent Ca2+ channel inhibitors, suggesting that these responses require extracellular Ca2+ influx via channels other than voltage-dependent Ca2+ channels. Antisense-mediated suppression of TRPV4 expression in intact cerebral arteries prevented 11,12 EET-induced smooth muscle hyperpolarization and vasodilation. Thus, we conclude that TRPV4 forms a novel Ca2+ signaling complex with ryanodine receptors and BKCa channels that elicits smooth muscle hyperpolarization and arterial dilation via Ca2+-induced Ca2+ release in response to an endothelial-derived factor. PMID- 16269660 TI - What is so special about apolipoprotein AI in reverse cholesterol transport? AB - An initial step in reverse cholesterol transport is the movement of unesterified cholesterol from peripheral cells to high-density lipoproteins (HDLs). This transfer usually occurs in extracellular spaces, such as the subendothelial space of a vessel wall, and is promoted by the interaction of lipid-free or lipid-poor apolipoprotein (apo)AI with ATP binding cassette A1 cellular transporters on macrophages (MPhi). Because HDL does not interact with MPhi ATP binding cassette A1 and apoAI is not synthesized by macrophages, this apoAI must be generated from spherical HDL. In this brief review, we propose that spherical apoAI is derived from HDL by remodeling events that are accomplished by proteins secreted by cholesteryl ester-loaded foam cells, including the lipid transfer proteins, phospholipid transfer protein, and cholesteryl ester transfer protein, and the triglyceride hydrolases hepatic lipase and lipoprotein lipase. PMID- 16269661 TI - Migration and growth are attenuated in vascular smooth muscle cells with type VIII collagen-null alleles. AB - OBJECTIVE: Type VIII collagen is upregulated after vascular injury and in atherosclerosis. However, the role of type VIII collagen endogenously expressed by smooth muscle cells (SMCs) and in the context of the vascular matrix microenvironment, which is rich in type I collagen, is not known. To address this, we have compared aortic SMCs from wild-type (WT) mice to SMCs from type VIII collagen-deficient (KO) mice when plated on type I collagen. METHODS AND RESULTS: Type VIII collagen was upregulated after wounding of WT SMCs. KO SMCs exhibited greater adhesion to type I collagen than WT SMCs (optical density [OD595]=0.458+/-0.044 versus 0.193+/-0.071). By contrast, the WT SMCs spread more (389+/-75% versus 108+/-14% increase in cell area), migrated further (total distance 80.6+/-6.2 microm versus 64.2+/-4.4 microm), and exhibited increased [3H]-thymidine uptake (160,000+/-22,300 versus 63,100+/-12,100 counts per minute) when compared with KO SMCs. Gelatin zymograms showed that WT SMCs expressed latent matrix metalloproteinase 2, whereas KO SMCs did not. Addition of exogenous type VIII collagen returned levels of KO SMC adhesion (OD595=0.316+/-0.038), migration (79.5+/-5.8 microm), and latent matrix metalloproteinase 2 expression to levels comparable to WT SMCs. CONCLUSIONS: This study suggests that SMCs can modify the matrix microenvironment by producing type VIII collagen, using it to overlay type I collagen, and generating a substrate favorable for migration. PMID- 16269662 TI - Interleukin 6 -174 g/c promoter polymorphism and risk of coronary heart disease: results from the rotterdam study and a meta-analysis. AB - OBJECTIVE: Inflammation plays a pivotal role in the pathogenesis of atherosclerosis. Interleukin (IL) 6 has many inflammatory functions, and the IL-6 -174 G/C promoter polymorphism appears to influence IL-6 levels. Findings of previous studies on the relation between this polymorphism and risk of cardiovascular diseases are inconsistent. We investigated this polymorphism in relation to risk of coronary heart disease (CHD) in a population-based study and meta-analysis. METHODS AND RESULTS: Participants (6434) of the Rotterdam Study were genotyped. Analyses on the relation between genotype and CHD were performed using Cox proportional hazards tests, and the association between genotype and plasma levels of IL-6 and C-reactive protein was investigated. All of the analyses were adjusted for age, sex, and common cardiovascular risk factors. A meta-analysis was performed, using a random effects model. No association between genotype and risk of CHD was observed. The polymorphism was not associated with IL-6 levels, but the C-allele was associated with higher C-reactive protein levels (P<0.01). Our meta-analysis did not show a significant association between the genotype and risk of CHD. CONCLUSIONS: We conclude that the polymorphism is not a suitable genetic marker for increased risk of CHD in subjects > or =55 years of age. PMID- 16269663 TI - Ferumoxtran-10-enhanced MRI of the hypercholesterolemic rabbit aorta: relationship between signal loss and macrophage infiltration. AB - OBJECTIVE: Ferumoxtran-10 is an MRI contrast agent, which accumulates in macrophages and induces magnetic susceptibility artifacts (MSAs). We evaluated the ability of ferumoxtran-10-enhanced MRI to quantify focal macrophage infiltration in the aortic wall of hypercholesterolemic rabbits. METHODS AND RESULTS: Six weeks after a double-balloon injury of the infrarenal aorta, 12 hypercholesterolemic rabbits underwent MRI of the aorta before (first MRI) and after (second MRI) intravenous injection of ferumoxtran-10 (n=10) or saline (n=2). A third MRI was performed 5 days later to detect ferumoxtran-10-induced MSA in the aortic wall. Aortas were subsequently processed for histology, immunohistochemistry, and gelatin zymography studies. Injured aortas displayed a macrophage-rich neointima with high-matrix metalloproteinase 2 and 9 activities. Iron stain of injured aortas showed massive accumulation of ferumoxtran-10 in neointimal macrophages. Five days after the injection of ferumoxtran-10, MSAs were detected only in the injured aortas by in vivo MRI and were quantified indirectly using the percentage reduction of luminal area attributable to the extension of these MSAs in the aortic lumen. This parameter correlated with macrophage infiltration on corresponding aortic cross-sections (r=0.82; P<0.05). CONCLUSIONS: Ferumoxtran-10-enhanced MRI allows quantitative assessment of macrophage infiltration induced by balloon angioplasty in the aorta of hypercholesterolemic rabbits. PMID- 16269664 TI - Cyclic strain-mediated regulation of vascular endothelial occludin and ZO-1: influence on intercellular tight junction assembly and function. AB - OBJECTIVE: The vascular endothelium constitutes a highly effective fluid/solute barrier through the regulated apposition of intercellular tight junction complexes. Because endothelium-mediated functions and pathology are driven by hemodynamic forces (cyclic strain and shear stress), we hypothesized a dynamic regulatory link between endothelial tight junction assembly/function and hemodynamic stimuli. We, therefore, examined the effects of cyclic strain on the expression, modification, and function of 2 pivotal endothelial tight junction components, occludin and ZO-1. METHODS AND RESULTS: For these studies, bovine aortic endothelial cells were subjected to physiological levels of equibiaxial cyclic strain (5% strain, 60 cycles/min, 24 hours). In response to strain, both occludin and ZO-1 protein expression increased by 2.3+/-0.1-fold and 2.0+/-0.3 fold, respectively, concomitant with a strain-dependent increase in occludin (but not ZO-1) mRNA levels. These changes were accompanied by reduced occludin tyrosine phosphorylation (75.7+/-8%) and increased ZO-1 serine/threonine phosphorylation (51.7+/-9% and 82.7+/-25%, respectively), modifications that could be completely blocked with tyrosine phosphatase and protein kinase C inhibitors (dephostatin and rottlerin, respectively). In addition, there was a significant strain-dependent increase in endothelial occludin/ZO-1 association (2.0+/-0.1-fold) in parallel with increased localization of both occludin and ZO 1 to the cell-cell border. These events could be completely blocked by dephostatin and rottlerin, and they correlated with a strain-dependent reduction in transendothelial permeability to FITC-dextran. CONCLUSIONS: Overall, these findings indicate that cyclic strain modulates both the expression and phosphorylation state of occludin and ZO-1 in vascular endothelial cells, with putative consequences for endothelial tight junction assembly and barrier integrity. PMID- 16269665 TI - Macrophage apolipoprotein E reduces atherosclerosis and prevents premature death in apolipoprotein E and scavenger receptor-class BI double-knockout mice. AB - OBJECTIVE: Mice null for both apolipoprotein (apo)E and scavenger receptor (SR) BI (DKO) develop severe hypercholesterolemia, occlusive coronary atherosclerosis, myocardial infarction, and premature death. The current study examines the ability of macrophage apoE to improve the dyslipidemia, reduce atherosclerosis, and rescue the lethal phenotype of DKO mice. METHODS AND RESULTS: Initially, bone marrow transplantation (BMT) was unsuccessful, because the DKO mice died from a rapidly fatal anemia 3 to 5 days after lethal irradiation. Therefore, probucol was used to rescue the DKO mice during BMT and was discontinued 2-weeks after BMT, allowing successful reconstitution with donor marrow. Twelve male apoE(-/ )SR-BI(-/-) mice fed 0.5% probucol in a chow diet were lethally irradiated and transplanted with either wild-type (WT) or DKO bone marrow. Two-weeks after BMT, apoE was detected in serum in WT-->DKO mice, and mean serum cholesterol levels were reduced by 70% versus DKO-->DKO mice. Lipoprotein profiles and HDL subpopulations in WT-->DKO mice were similar to apoE(+/+)SR-BI(-/-)-->DKO mice and resembled those of SR-BI(-/-) mice. In WT-->DKO mice, aortic atherosclerosis was reduced by 88% to 90% versus DKO-->DKO mice. Furthermore, the DKO-->DKO mice died &8 weeks after BMT, whereas WT-->DKO mice exhibited a life span >40 weeks after BMT. CONCLUSIONS: Macrophage apoE is able to rescue the lethal phenotype of apoE(-/-)SR-BI(-/-) mice by improving the dyslipidemia and dramatically reducing atherosclerotic lesion development. PMID- 16269666 TI - Coronary artery atherosclerosis is related to reduced regional left ventricular function in individuals without history of clinical cardiovascular disease: the Multiethnic Study of Atherosclerosis. AB - OBJECTIVE: We investigated whether regional coronary calcium score by computed tomography is related to regional left ventricular systolic function measured by MRI tagging in participants of the Multiethnic Study of Atherosclerosis. METHODS AND RESULTS: The Multiethnic Study of Atherosclerosis is a prospective observational study of men and women without a history of previous heart disease from 4 ethnic groups. Calcium scores were measured separately for the left anterior descendent (LAD), left circumflex (LCX), and right (RCA) coronary arteries. Left ventricular strain and strain rate were determined by tagged MRI in the corresponding vascular territories of the coronary vessels in 509 participants. Greater coronary calcification in the LAD, LCX, and right RCA coronary arteries were related to worse function in their respective perfusion. Anterior wall strain rate was -1.37+/-0.41 when LAD calcium was zero versus 1.17+/-0.24 1/s in the highest quartile of calcium score (P<0.001). Similar relationships were evident in the LCX and RCA regions. Participants with 1- and 2 vessel coronary artery calcium had better myocardial function in the remote area compared with the territory supplied by the diseased artery. CONCLUSIONS: High local calcium score is related to regional dysfunction in the corresponding coronary territory among individuals without a history of previous heart disease. These results indicate a link between atherosclerosis and subclinical regional left ventricular dysfunction. PMID- 16269667 TI - Adiponectin acts as an endogenous antithrombotic factor. AB - OBJECTIVE: Obesity is a common risk factor in insulin resistance and cardiovascular diseases. Although hypoadiponectinemia is associated with obesity related metabolic and vascular diseases, the role of adiponectin in thrombosis remains elusive. METHODS AND RESULTS: We investigated platelet thrombus formation in adiponectin knockout (APN-KO) male mice (8 to 12 weeks old) fed on a normal diet. There was no significant difference in platelet counts or coagulation parameters between wild-type (WT) and APN-KO mice. However, APN-KO mice showed an accelerated thrombus formation on carotid arterial injury with a He-Ne laser (total thrombus volume: 13.36+/-4.25 x 10(7) arbitrary units for APN-KO and 6.74+/-2.87x10(7) arbitrary units for WT; n=10; P<0.01). Adenovirus-mediated supplementation of adiponectin attenuated the enhanced thrombus formation. In vitro thrombus formation on a type I collagen at a shear rate of 250 s(-1), as well as platelet aggregation induced by low concentrations of agonists, was enhanced in APN-KO mice, and recombinant adiponectin inhibited the enhanced platelet aggregation. In WT mice, adenovirus-mediated overexpression of adiponectin additionally attenuated thrombus formation. CONCLUSIONS: Adiponectin deficiency leads to enhanced thrombus formation and platelet aggregation. The present study reveals a new role of adiponectin as an endogenous antithrombotic factor. PMID- 16269668 TI - Endothelial nitric oxide synthase activation by tumor necrosis factor alpha through neutral sphingomyelinase 2, sphingosine kinase 1, and sphingosine 1 phosphate receptors: a novel pathway relevant to the pathophysiology of endothelium. AB - OBJECTIVE: Tumor necrosis factor alpha (TNF-alpha), a key proinflammatory cytokine acting on the endothelium, activates endothelial nitric oxide synthase (eNOS). We have examined the signaling pathway leading to this activation and its biological role in endothelium, which are still unknown. METHODS AND RESULTS: In human endothelial cells, we found that eNOS activation by TNF-alpha is time dependent and requires activation of Akt, a known eNOS activator. eNOS activation was preceded by sequential activation of neutral-sphingomyelinase-2 (N-SMase2) and sphingosine-kinase-1 (SK1) and generation of sphingosine-1-phosphate (Sph1P). Inhibition of N-SMase2 inhibited Sph1P formation, whereas inhibition of SK1 did not affect N-SMase2 activation by TNF-alpha. Blockade of N-SMase2, SK1, or the Sph1P receptors S1P1 and S1P3, either by silencing or pharmacological inhibitors, prevented eNOS activation. Thus, eNOS is activated by TNF-alpha via S1P receptors, activated by Sph1P generated through N-SMase2 and SK1 activation. We found that nitric oxide generated through this pathway has a biological role, because it inhibits the expression of E-selectin and the adhesion of dendritic cells to the endothelium stimulated by TNF-alpha. CONCLUSIONS: This study establishes a previously undescribed link among TNF-alpha, Sph1P, and eNOS in a same signaling pathway of biological relevance in the process of endothelial cell activation by TNF-alpha. PMID- 16269669 TI - Ritonavir impairs lipoprotein lipase-mediated lipolysis and decreases uptake of fatty acids in adipose tissue. AB - OBJECTIVE: The use of the HIV protease inhibitor ritonavir (RTV) is frequently associated with hypertriglyceridemia and lipodystrophy. The aim of our study was to determine the mechanism underlying the observed hypertriglyceridemia. METHODS AND RESULTS: Feeding female APOE*3-Leiden transgenic mice a Western-type diet supplemented with RTV (35 mg/kg per day) for 2 weeks resulted in a 2-fold increase in fasting plasma triglyceride (TG) levels, which was specific for very low-density lipoprotein (VLDL). RTV did not change the hepatic VLDL-TG production. Instead, RTV did increase the postprandial TG response to an oral fat load (area under the curve, 25.5+/-12.1 versus 13.8+/-6.8 mmol/L per hour in controls; P<0.05). Likewise, RTV hampered the plasma clearance of intravenously injected glycerol tri[3H]oleate-labeled VLDL-like emulsion particles (half time, 19.3+/-10.5 versus 5.0+/-1.3 minutes in controls; P<0.05) associated with a decrease of 44% in plasma lipoprotein lipase activity. Accordingly, RTV decreased the uptake of TG-derived fatty acids (FAs) into adipose tissue, as well as the uptake of albumin-bound FA. CONCLUSIONS: We conclude that RTV causes hypertriglyceridemia via decreased lipoprotein lipase-mediated clearance of VLDL TG. In addition, RTV specifically impairs the uptake of FA in adipose tissue, which may contribute to the lipodystrophy that is frequently observed in HIV infected subjects on antiretroviral therapy. PMID- 16269671 TI - Diversity of "Candidatus Liberibacter asiaticus," based on the omp gene sequence. AB - Huanglongbing (yellow dragon disease) is a destructive disease of citrus. The etiological agent is a noncultured, phloem-restricted alpha-proteobacterium, "Candidatus Liberibacter africanus" in Africa and "Candidatus Liberibacter asiaticus" in Asia. In this study, we used an omp-based PCR-restriction fragment length polymorphism (RFLP) approach to analyze the genetic variability of "Ca. Liberibacter asiaticus" isolates. By using five different enzymes, each the 10 isolates tested could be associated with a specific combination of restriction profiles. The results indicate that the species "Ca. Liberibacter asiaticus," even within a given region, may comprise several different variants. Thus, omp based PCR-RFLP analysis is a simple method for detecting and differentiating "Ca. Liberibacter asiaticus" isolates. PMID- 16269670 TI - Improvement of galactose uptake in Saccharomyces cerevisiae through overexpression of phosphoglucomutase: example of transcript analysis as a tool in inverse metabolic engineering. AB - Through genome-wide transcript analysis of a reference strain and two recombinant Saccharomyces cerevisiae strains with different rates of galactose uptake, we obtained information about the global transcriptional response to metabolic engineering of the GAL gene regulatory network. One of the recombinant strains overexpressed the gene encoding the transcriptional activator Gal4, and in the other strain the genes encoding Gal80, Gal6, and Mig1, which are negative regulators of the GAL system, were deleted. Even though the galactose uptake rates were significantly different in the three strains, we surprisingly did not find any significant changes in the expression of the genes encoding the enzymes catalyzing the first steps of the pathway (i.e., the genes encoding Gal2, Gal1, Gal7, and Gal10). We did, however, find that PGM2, encoding the major isoenzyme of phosphoglucomutase, was slightly up-regulated in the two recombinant strains with higher galactose uptake rates. This indicated that PGM2 is a target for overexpression in terms of increasing the flux through the Leloir pathway, and through overexpression of PGM2 the galactose uptake rate could be increased by 70% compared to that of the reference strain. Based on our findings, we concluded that phosphoglucomutase plays a key role in controlling the flux through the Leloir pathway, probably due to increased conversion of glucose-1-phosphate to glucose-6-phosphate. This conclusion was supported by measurements of sugar phosphates, which showed that there were increased concentrations of glucose-6 phosphate, galactose-6-phosphate, and fructose-6-phosphate in the strain construct overexpressing PGM2. PMID- 16269672 TI - Culture-independent techniques for rapid detection of bacteria associated with loss of chloramine residual in a drinking water system. AB - Chloramination is often the disinfection regimen of choice for extended drinking water systems. However, this process is prone to instability due to the growth of nitrifying bacteria. This is the first study to use alternative approaches for rapid investigation of chloraminated drinking water system instability in which flow cytometric cell sorting of bacteria with intact membranes (membrane-intact fraction) (BacLight kit) or with active esterases (esterase-active fraction) (carboxyfluorescein diacetate) was combined with 16S rRNA gene-directed PCR and denaturing gradient gel electrophoresis (DGGE). No active bacteria were detected when water left the water treatment plant (WTP), but 12 km downstream the chloramine residual had diminished and the level of active bacteria in the bulk water had increased to more than 1 x 10(5) bacteria ml(-1). The bacterial diversity in the system was represented by six major DGGE bands for the membrane intact fraction and 10 major DGGE bands for the esterase-active fraction. PCR targeting of the 16S rRNA gene of chemolithotrophic ammonia-oxidizing bacteria (AOB) and subsequent DGGE and DNA sequence analysis revealed the presence of an active Nitrosospira-related species and Nitrosomonas cryotolerans in the system, but no AOB were detected in the associated WTP. The abundance of active AOB was then determined by quantitative real-time PCR (qPCR) targeting the amoA gene; 3.43 x 10(3) active AOB ml(-1) were detected in the membrane-intact fraction, and 1.40 x 10(4) active AOB ml(-1) were detected in the esterase-active fraction. These values were several orders of magnitude greater than the 2.5 AOB ml(-1) detected using a routine liquid most-probable-number assay. Culture-independent techniques described here, in combination with existing chemical indicators, should allow the water industry to obtain more comprehensive data with which to make informed decisions regarding remedial action that may be required either prior to or during an instability event. PMID- 16269673 TI - Surface microflora of four smear-ripened cheeses. AB - The microbial composition of smear-ripened cheeses is not very clear. A total of 194 bacterial isolates and 187 yeast isolates from the surfaces of four Irish farmhouse smear-ripened cheeses were identified at the midpoint of ripening using pulsed-field gel electrophoresis (PFGE), repetitive sequence-based PCR, and 16S rRNA gene sequencing for identifying and typing the bacteria and Fourier transform infrared spectroscopy and mitochondrial DNA restriction fragment length polymorphism (mtDNA RFLP) analysis for identifying and typing the yeast. The yeast microflora was very uniform, and Debaryomyces hansenii was the dominant species in the four cheeses. Yarrowia lipolytica was also isolated in low numbers from one cheese. The bacteria were highly diverse, and 14 different species, Corynebacterium casei, Corynebacterium variabile, Arthrobacter arilaitensis, Arthrobacter sp., Microbacterium gubbeenense, Agrococcus sp. nov., Brevibacterium linens, Staphylococcus epidermidis, Staphylococcus equorum, Staphylococcus saprophyticus, Micrococcus luteus, Halomonas venusta, Vibrio sp., and Bacillus sp., were identified on the four cheeses. Each cheese had a more or less unique microflora with four to nine species on its surface. However, two bacteria, C. casei and A. arilaitensis, were found on each cheese. Diversity at the strain level was also observed, based on the different PFGE patterns and mtDNA RFLP profiles of the dominant bacterial and yeast species. None of the ripening cultures deliberately inoculated onto the surface were reisolated from the cheeses. This study confirms the importance of the adventitious, resident microflora in the ripening of smear cheeses. PMID- 16269674 TI - Isolation and partial characterization of antagonistic peptides produced by Paenibacillus sp. strain B2 isolated from the sorghum mycorrhizosphere. AB - Paenibacillus sp. strain B2, isolated from the mycorrhizosphere of sorghum colonized by Glomus mosseae, produces an antagonistic factor. This factor has a broad spectrum of activity against gram-positive and gram-negative bacteria and also against fungi. The antagonistic factor was isolated from the bacterial culture medium and purified by cation-exchange, reverse-phase, and size exclusion chromatography. The purified factor could be separated into three active compounds following characterization by amino acid analysis and by combined reverse-phase chromatography and mass spectrometry (liquid chromatography-mass spectrometry and mass spectrometry-mass spectrometry). The first compound had the same retention time as polymyxin B1, whereas the two other compounds were more hydrophobic. The molecular masses of the latter compounds are 1,184.7 and 1,202.7 Da, respectively, and their structure is similar to that of polymyxin B1, with a cyclic heptapeptide moiety attached to a tripeptide side chain and a fatty acyl residue. They both contain threonine, phenylalanine, leucine, and 2,4 diaminobutyric acid residues. The peptide with a molecular mass of 1,184.7 contains a 2,3-didehydrobutyrine residue with a molecular mass of 101 Da replacing a threonine at the A2 position of the polymyxin side chain. This modification could explain the broader range of antagonistic activity of this peptide compared to that of polymyxin B. PMID- 16269675 TI - Characterization of enzymatic antioxidants in the lichen Ramalina lacera and their response to rehydration. AB - Lichens are slow-growing associations of fungi and green algae or cyanobacteria. This symbiotic association forms a common thallus that does not possess roots or a waxy cuticle and depends mainly on atmospheric input of mineral nutrients. The lifestyle of most lichens is composed of alternating periods of desiccation with low metabolic activity and hydration that induces increase in their metabolism. We have previously shown that rehydration of the naturally desiccated lichen Ramalina lacera resulted in a rapid increase in photosynthesis and was accompanied by a burst of intracellular production of reactive oxygen species and nitric oxide, as well as a transient decrease in water-soluble antioxidant capacity. We report here on enzymatic antioxidants of R. lacera and their response to rehydration. Native gel electrophoresis of crude extracts of R. lacera stained for superoxide dismutase (SOD) activity revealed four Fe-SOD and four Mn-SOD electromorphs that are synthesized by the alga, a Cu/Zn-SOD and a Mn SOD that are the product of the fungus, and two catalases synthesized one by the fungus and the other by the algae. In addition, we detected glutathione reductase and glucose-6-phosphate dehydrogenase activities in crude extracts of R. lacera. Rehydration of the thalli resulted in a decrease in SOD activity of all forms, and a transient decrease in total catalase activity, as well as a decrease in the antioxidant auxiliary enzymes glutathione reductase and glucose-6-phosphate dehydrogenase. PMID- 16269676 TI - CorA affects tolerance of Escherichia coli and Salmonella enterica serovar Typhimurium to the lactoperoxidase enzyme system but not to other forms of oxidative stress. AB - The enzyme lactoperoxidase is part of the innate immune system in vertebrates and owes its antimicrobial activity to the formation of oxidative reaction products from various substrates. In a previous study, we have reported that, with thiocyanate as a substrate, the lactoperoxidase system elicits a distinct stress response in Escherichia coli MG1655. This response is different from but partly overlapping with the stress responses to hydrogen peroxide and to superoxide. In the current work, we constructed knockouts in 10 lactoperoxidase system-inducible genes to investigate their role in the tolerance of E. coli MG1655 to this antimicrobial system. Five mutations resulted in a slightly increased sensitivity, but one mutation (corA) caused hypersensitivity to the lactoperoxidase system. This hypersensitive phenotype was specific to the lactoperoxidase system, since neither the sensitivity to hydrogen peroxide nor to the superoxide generator plumbagin was affected in the corA mutant. Salmonella enterica serovar Typhimurium corA had a similar phenotype. Although corA encodes an Mg2+ transporter and at least three other inducible open reading frames belonged to the Mg2+ regulon, repression of the Mg stimulon by Mg2+ did not change the lactoperoxidase sensitivity of either the wild-type or corA mutant. Prior exposure to 0.3 mM Ni2+, which is also transported by CorA, strongly sensitized MG1655 but not the corA mutant to the lactoperoxidase system. Furthermore, this Ni2+-dependent sensitization was suppressed by the CorA specific inhibitor Co(III) hexaammine. These results indicate that CorA affects the lactoperoxidase sensitivity of E. coli by modulating the cytoplasmic concentrations of transition metals that enhance the toxicity of the lactoperoxidase system. PMID- 16269678 TI - Lactobacillus paracasei subsp. paracasei 8700:2 degrades inulin-type fructans exhibiting different degrees of polymerization. AB - Ten strains of lactobacilli were assessed for their capacity to degrade inulin type fructans, which are well-known prebiotics. Both oligofructose and inulin were tested. The dairy isolate Lactobacillus acidophilus IBB 801 degraded only oligofructose. The human isolate Lactobacillus paracasei subsp. paracasei 8700:2 degraded oligofructose and long-chain inulin and grew rapidly on both energy sources. In both cases, fractions of different degrees of polymerization were fermented. Moreover, large and short fractions of oligofructose were degraded simultaneously. When L. paracasei subsp. paracasei 8700:2 grew on oligofructose enriched inulin, oligofructose was preferentially metabolized. In all cases, lactic acid was the main metabolic end product. Significant amounts of acetic acid, formic acid, and ethanol were produced when long-chain inulin or oligofructose-enriched inulin was used as the sole energy source. PMID- 16269677 TI - Differentiation of spores of Bacillus subtilis grown in different media by elemental characterization using time-of-flight secondary ion mass spectrometry. AB - We demonstrate the use of time-of-flight secondary ion mass spectrometry (TOF SIMS) in a forensics application to distinguish Bacillus subtilis spores grown in various media based on the elemental signatures of the spores. Triplicate cultures grown in each of four different media were analyzed to obtain TOF-SIMS signatures comprised of 16 elemental intensities. Analysis of variance was unable to distinguish growth medium types based on 40Ca-normalized signatures of any single normalized element. Principal component analysis proved successful in separating the spores into groups consistent with the media in which they were prepared. Confusion matrices constructed using nearest-neighbor classification of the PCA scores confirmed the predictive utility of TOF-SIMS elemental signatures in identifying sporulation medium. Theoretical calculations based on the number and density of spores in an analysis area indicate an analytical sample size of about 1 ng, making this technique an attractive method for bioforensics applications. PMID- 16269679 TI - Novel 4-chlorophenol degradation gene cluster and degradation route via hydroxyquinol in Arthrobacter chlorophenolicus A6. AB - Arthrobacter chlorophenolicus A6, a previously described 4-chlorophenol-degrading strain, was found to degrade 4-chlorophenol via hydroxyquinol, which is a novel route for aerobic microbial degradation of this compound. In addition, 10 open reading frames exhibiting sequence similarity to genes encoding enzymes involved in chlorophenol degradation were cloned and designated part of a chlorophenol degradation gene cluster (cph genes). Several of the open reading frames appeared to encode enzymes with similar functions; these open reading frames included two genes, cphA-I and cphA-II, which were shown to encode functional hydroxyquinol 1,2-dioxygenases. Disruption of the cphA-I gene yielded a mutant that exhibited negligible growth on 4-chlorophenol, thereby linking the cph gene cluster to functional catabolism of 4-chlorophenol in A. chlorophenolicus A6. The presence of a resolvase pseudogene in the cph gene cluster together with analyses of the G+C content and codon bias of flanking genes suggested that horizontal gene transfer was involved in assembly of the gene cluster during evolution of the ability of the strain to grow on 4-chlorophenol. PMID- 16269680 TI - Isolation and characterization of a novel antialgal allelochemical from Phragmites communis. AB - Antialgal allelochemicals were isolated from Phragmites communis Tris. The isolated allelopathic fraction showed strong inhibition activity on the growth of Chlorella pyrenoidosa and Microcystis aeruginosa but had no inhibition on Chlorella vulgaris. The 50% effective concentrations (EC50) of the allelopathic fractions on C. pyrenoidosa and M. aeruginosa were 0.49 and 0.79 mg/liter, respectively. The allelopathic activity of the fraction was species-specific. The isolated allelopathic fraction caused metal ion leakage from algal cells. The fraction decreased the activities of antioxidant enzymes, such as superoxide dismutase and peroxidase. The addition of the isolated fraction increased the concentration of unsaturated lipid fatty acids in cell membrane of C. pyrenoidosa and M. aeruginosa. This caused a change in plasma membrane integrity and the leakage of ions in the protoplast. The allelopathic compound was identified by nuclear magnetic resonance and gas chromatography-mass spectrometry as ethyl 2 methylacetoacetate. Synthesized ethyl 2-methylacetoacetate also showed allelopathic activity on C. pyrenoidosa and M. aeruginosa. The EC50 of synthesized ethyl 2-methylacetoacetate on C. pyrenoidosa and M. aeruginosa were 0.49 and 0.65 mg/liter, respectively. PMID- 16269681 TI - Bacteriophage therapy to reduce Campylobacter jejuni colonization of broiler chickens. AB - Colonization of broiler chickens by the enteric pathogen Campylobacter jejuni is widespread and difficult to prevent. Bacteriophage therapy is one possible means by which this colonization could be controlled, thus limiting the entry of campylobacters into the human food chain. Prior to evaluating the efficacy of phage therapy, experimental models of Campylobacter colonization of broiler chickens were established by using low-passage C. jejuni isolates HPC5 and GIIC8 from United Kingdom broiler flocks. The screening of 53 lytic bacteriophage isolates against a panel of 50 Campylobacter isolates from broiler chickens and 80 strains isolated after human infection identified two phage candidates with broad host lysis. These phages, CP8 and CP34, were orally administered in antacid suspension, at different dosages, to 25-day-old broiler chickens experimentally colonized with the C. jejuni broiler isolates. Phage treatment of C. jejuni colonized birds resulted in Campylobacter counts falling between 0.5 and 5 log10 CFU/g of cecal contents compared to untreated controls over a 5-day period postadministration. These reductions were dependent on the phage-Campylobacter combination, the dose of phage applied, and the time elapsed after administration. Campylobacters resistant to bacteriophage infection were recovered from phage-treated chickens at a frequency of <4%. These resistant types were compromised in their ability to colonize experimental chickens and rapidly reverted to a phage-sensitive phenotype in vivo. The selection of appropriate phage and their dose optimization are key elements for the success of phage therapy to reduce campylobacters in broiler chickens. PMID- 16269682 TI - A bile salt-resistant derivative of Bifidobacterium animalis has an altered fermentation pattern when grown on glucose and maltose. AB - The growth of Bifidobacterium animalis subsp. lactis IPLA 4549 and its derivative with acquired resistance to bile, B. animalis subsp. lactis 4549dOx, was evaluated in batch cultures with glucose or the glucose disaccharide maltose as the main carbon source. The acquisition of bile salt resistance caused a change in growth pattern for both sugars, which mainly resulted in a preferential use of maltose compared to glucose, whereas the mother strain used both carbohydrates in a similar way. High-performance liquid chromatography and gas chromatography-mass spectrometry analyses were performed to determine the amounts of glucose consumption and organic acid and ethanol formation from glucose by buffered resting cells taken at different points during growth. Resting cells of the bile adapted strain generally consumed less glucose than those of the nonadapted one but showed an enhanced production of ethanol and higher acetic acid-to-lactic acid as well as formic acid-to-lactic acid ratios. These findings suggest a shift in the catabolism of carbohydrates promoted by the acquisition of bile resistance that may cause changes in the redox potential and improvements in the cellular ATP yield. PMID- 16269683 TI - Importance of organosulfur utilization for survival of Pseudomonas putida in soil and rhizosphere. AB - The sulfur present in both agricultural and uncultivated soils is largely in the form of sulfonates and sulfate esters and not as free, bioavailable inorganic sulfate. Desulfurization of the former compounds in vitro has previously been studied in Pseudomonas putida, a common rhizosphere inhabitant. Survival of P. putida strains was now investigated in three sulfur-deficient Danish soils which were found to contain 60 to 70% of their sulfur in sulfonate or sulfate ester form, as determined by X-ray near-edge spectroscopy. The soil fitness of P. putida S-313 was compared with that of isogenic strains with mutations in the sftR and asfA genes (required for in vitro desulfurization of sulfate esters and arylsulfonates, respectively) and in the ssu locus (required in vitro for the desulfurization of both sulfonates and sulfate esters). asfA or sftR mutants showed significantly reduced survival compared to the parent strain in bulk soil that had been enriched with carbon and nitrogen to mimic rhizosphere conditions, but this reduced survival was not observed in the absence of these additives. In a tomato rhizosphere grown in compost, survival of sftR and ssu mutants was reduced relative to the parent strain. The results demonstrate that the ability to desulfurize sulfonates and sulfate esters is critical for survival of bacteria in the rhizosphere but less so in bulk soils outside the influence of plant roots, where carbon is the limiting nutrient for growth. PMID- 16269684 TI - Preparation, characterization, and optimization of an in vitro C30 carotenoid pathway. AB - The ispA gene encoding farnesyl pyrophosphate (FPP) synthase from Escherichia coli and the crtM gene encoding 4,4'-diapophytoene (DAP) synthase from Staphylococcus aureus were overexpressed and purified for use in vitro. Steady state kinetics for FPP synthase and DAP synthase, individually and in sequence, were determined under optimized reaction conditions. For the two-step reaction, the DAP product was unstable in aqueous buffer; however, in situ extraction using an aqueous-organic two-phase system resulted in a 100% conversion of isopentenyl pyrophosphate and dimethylallyl pyrophosphate into DAP. This aqueous-organic two phase system is the first demonstration of an in vitro carotenoid synthesis pathway performed with in situ extraction, which enables quantitative conversions. This approach, if extended to a wide range of isoprenoid-based pathways, could lead to the synthesis of novel carotenoids and their derivatives. PMID- 16269685 TI - Genetic and phylogeographic structures of the symbiotic fungus Tuber magnatum. AB - The quality and market price of truffles vary with the species and, traditionally, the place of origin. The premium species Tuber magnatum produces white truffles and has a patchy distribution restricted to Italy and some Balkan areas. We used polymorphic microsatellites to evaluate 316 specimens grouped into 26 populations sampled across the species' geographic range to determine if natural populations of T. magnatum are genetically differentiated. We found that the southernmost and the northwesternmost populations were significantly differentiated from the rest of the populations. The simple sequence repeat data also could be used to make inferences about the postglacial T. magnatum expansion pattern. This study is the first to identify a genetic and phylogeographic structure in T. magnatum. The presence of a genetic structure can be of practical interest in tracing truffle populations according to their geographic origin for marketing strategies. Evidence for extensive outcrossing in field populations of T. magnatum also is provided for the first time. PMID- 16269687 TI - The halophilic fungus Hortaea werneckii and the halotolerant fungus Aureobasidium pullulans maintain low intracellular cation concentrations in hypersaline environments. AB - Hortaea werneckii and Aureobasidium pullulans, black yeast-like fungi isolated from hypersaline waters of salterns as their natural ecological niche, have been previously defined as halophilic and halotolerant microorganisms, respectively. In the present study we assessed their growth and determined the intracellular cation concentrations of salt-adapted and non-salt-adapted cells of both species at a wide range of salinities (0 to 25% NaCl and 0 to 20% NaCl, respectively). Although 5% NaCl improved the growth of H. werneckii, even the minimal addition of NaCl to the growth medium slowed down the growth rate of A. pullulans, confirming their halophilic and halotolerant nature. Salt-adapted cells of H. werneckii and A. pullulans kept very low amounts of internal Na+ even when grown at high NaCl concentrations and can be thus considered Na+ excluders, suggesting the existence of efficient mechanisms for the regulation of ion fluxes. Based on our results, we can conclude that these organisms do not use K+ or Na+ for osmoregulation. Comparison of cation fluctuations after a hyperosmotic shock, to which nonadapted cells of both species were exposed, demonstrated better ionic homeostasis regulation of H. werneckii compared to A. pullulans. We observed small fluctuations of cation concentrations after a hyperosmotic shock in nonadapted A. pullulans similar to those in salt-adapted H. werneckii, which additionally confirmed better regulation of ionic homeostasis in the latter. These features can be expected from organisms adapted to survival within a wide range of salinities and to occasional exposure to extremely high NaCl concentrations, both characteristic for their natural environment. PMID- 16269688 TI - Prediction of the adaptability of Pseudomonas putida DOT-T1E to a second phase of a solvent for economically sound two-phase biotransformations. AB - The strain Pseudomonas putida DOT-T1E was tested for its ability to tolerate second phases of different alkanols for their use as solvents in two-liquid-phase biotransformations. Although 1-decanol showed an about 10-fold higher toxicity to the cells than 1-octanol, the cells were able to adapt completely to 1-decanol only and could not be adapted in order to grow stably in the presence of a second phase of 1-octanol. The main explanation for this observation can be seen in the higher water and membrane solubility of 1-octanol. The hydrophobicity (log P) of a substance correlates with a certain partitioning of that compound into the membrane. Combining the log P value with the water solubility, the maximum membrane concentration of a compound can be calculated. With this simple calculation, it is possible to predict the property of an organic chemical for its potential applicability as a solvent for two-liquid-phase biotransformations with solvent-tolerant P. putida strains. Only compounds that show a maximum membrane concentration of less than 400 mM, such as 1-decanol, seem to be tolerated by these bacterial strains when applied in supersaturating concentrations to the medium. Taking into consideration that a solvent for a two liquid-phase system should possess partitioning properties for potential substrates and products of a fine chemical synthesis, it can be seen that 1 decanol is a suitable solvent for such biotransformation processes. This was also demonstrated in shake cultures, where increasing amounts of a second phase of 1 decanol led to bacteria tolerating higher concentrations of the model substrate 3 nitrotoluene. Transferring this example to a 5-liter-scale bioreactor with 10% (vol/vol) 1-decanol, the amount of 3-nitrotoluene tolerated by the cells is up to 200-fold higher than in pure aqueous medium. The system demonstrates the usefulness of two-phase biotransformations utilizing solvent-tolerant bacteria. PMID- 16269686 TI - Intra- and interspecific comparisons of bacterial diversity and community structure support coevolution of gut microbiota and termite host. AB - We investigated the bacterial gut microbiota from 32 colonies of wood-feeding termites, comprising four Microcerotermes species (Termitidae) and four Reticulitermes species (Rhinotermitidae), using terminal restriction fragment length polymorphism analysis and clonal analysis of 16S rRNA. The obtained molecular community profiles were compared statistically between individuals, colonies, locations, and species of termites. Both analyses revealed that the bacterial community structure was remarkably similar within each termite genus, with small but significant differences between sampling sites and/or termite species. In contrast, considerable differences were found between the two termite genera. Only one bacterial phylotype (defined with 97% sequence identity) was shared between the two termite genera, while 18% and 50% of the phylotypes were shared between two congeneric species in the genera Microcerotermes and Reticulitermes, respectively. Nevertheless, a phylogenetic analysis of 228 phylotypes from Microcerotermes spp. and 367 phylotypes from Reticulitermes spp. with other termite gut clones available in public databases demonstrated the monophyly of many phylotypes from distantly related termites. The monophyletic "termite clusters" comprised of phylotypes from more than one termite species were distributed among 15 bacterial phyla, including the novel candidate phyla TG2 and TG3. These termite clusters accounted for 95% of the 960 clones analyzed in this study. Moreover, the clusters in 12 phyla comprised phylotypes from more than one termite (sub)family, accounting for 75% of the analyzed clones. Our results suggest that the majority of gut bacteria are not allochthonous but are specific symbionts that have coevolved with termites and that their community structure is basically consistent within a genus of termites. PMID- 16269690 TI - Suitability of recombinant Escherichia coli and Pseudomonas putida strains for selective biotransformation of m-nitrotoluene by xylene monooxygenase. AB - Escherichia coli JM101(pSPZ3), containing xylene monooxygenase (XMO) from Pseudomonas putida mt-2, catalyzes specific oxidations and reductions of m nitrotoluene and derivatives thereof. In addition to reactions catalyzed by XMO, we focused on biotransformations by native enzymes of the E. coli host and their effect on overall biocatalyst performance. While m-nitrotoluene was consecutively oxygenated to m-nitrobenzyl alcohol, m-nitrobenzaldehyde, and m-nitrobenzoic acid by XMO, the oxidation was counteracted by an alcohol dehydrogenase(s) from the E. coli host, which reduced m-nitrobenzaldehyde to m-nitrobenzyl alcohol. Furthermore, the enzymatic background of the host reduced the nitro groups of the reactants resulting in the formation of aromatic amines, which were shown to effectively inhibit XMO in a reversible fashion. Host-intrinsic oxidoreductases and their reaction products had a major effect on the activity of XMO during biocatalysis of m-nitrotoluene. P. putida DOT-T1E and P. putida PpS81 were compared to E. coli JM101 as alternative hosts for XMO. These promising strains contained an additional dehydrogenase that oxidized m-nitrobenzaldehyde to the corresponding acid but catalyzed the formation of XMO-inhibiting aromatic amines at a significantly lower level than E. coli JM101. PMID- 16269689 TI - Comparison of minisatellite polymorphisms in the Bacillus cereus complex: a simple assay for large-scale screening and identification of strains most closely related to Bacillus anthracis. AB - Polymorphism of five tandem repeats that are monomorphic in Bacillus anthracis was investigated in 230 isolates of the B. cereus group and in 5 sequenced B. cereus genomes in search for markers allowing identification of B. cereus and B. thuringiensis strains most closely related to B. anthracis. Using this multiple locus variable number of tandem repeat analysis (MLVA), a cluster of 30 strains was selected for further characterization. Eventually, six of these were characterized by multilocus sequence type analysis. One of the strains is only six point mutations (of almost 3,000 bp) away from B. anthracis and was also proposed to be closest to B. anthracis by MLVA analysis. However, this strain remains separated from B. anthracis by a number of significant genetic events observed in B. anthracis, including the loss of the hemolysin activity, the presence of four prophages, and the presence of the two virulence plasmids, pXO1 and pXO2. One particular minisatellite marker provides an efficient assay to identify the subset of B. cereus and B. thuringiensis strains closely related to B. anthracis. Based on these results, a very simple assay is proposed that allows the screening of hundreds of strains from the B. cereus complex, with modest equipment and at a low cost, to eventually fill the gap with B. anthracis and better understand the origin and making of this dangerous pathogen. PMID- 16269692 TI - Viral production, decay rates, and life strategies along a trophic gradient in the North Adriatic Sea. AB - Although the relationships between trophic conditions and viral dynamics have been widely explored in different pelagic environments, there have been few attempts at independent estimates of both viral production and decay. In this study, we investigated factors controlling the balance between viral production and decay along a trophic gradient in the north Adriatic basin, providing independent estimates of these variables and determining the relative importance of nanoflagellate grazing and viral life strategies. Increasing trophic conditions induced an increase of bacterioplankton growth rates and of the burst sizes. As a result, eutrophic waters displayed highest rates of viral production, which considerably exceeded observed rates of viral decay (up to 2.9 x 10(9) VLP liter(-1) h(-1)). Viral decay was also higher in eutrophic waters, where it accounted for ca. 40% of viral production, and dropped significantly to 1.3 to 10.7% in oligotrophic waters. These results suggest that viral production and decay rates may not necessarily be balanced in the short term, resulting in a net increase of viruses in the system. In eutrophic waters nanoflagellate grazing, dissolved-colloidal substances, and lysogenic infection were responsible together for the removal of ca. 66% of viral production versus 17% in oligotrophic waters. Our results suggest that different causative agents are primarily responsible for the removal of viruses from the water column in different trophic conditions. Factors other than those considered in the past might shed light on processes responsible for the removal and/or decay of viral particles from the water column. PMID- 16269691 TI - Detection, characterization, and in vitro and in vivo expression of genes encoding S-proteins in Lactobacillus gallinarum strains isolated from chicken crops. AB - Thirty-eight isolates of Lactobacillus gallinarum cultured from the crops of broiler chickens were screened for the presence of genes encoding S-layer proteins. All of the isolates had two S-protein genes, which were designated Lactobacillus gallinarum S-protein (lgs) genes. One gene in each isolate was either lgsA or lgsB. The Lactobacillus isolates were further characterized by pulsed-field gel electrophoresis of DNA digests, which grouped the isolates into 17 genotypes (strains). The second gene in each of eight representative strains was sequenced and shown to differ among strains (lgsC, lgsD, lgsE, lgsF, lgsG, lgsH, and lgsI). The genome of each strain thus encoded a common S-protein (encoded by either lgsA or lgsB) and a strain-specific S-protein. The extraction of cell surface proteins from cultures of the eight strains showed that each strain produced a single S-protein that was always encoded by the strain-specific lgs gene. Two of the strains were used to inoculate chickens maintained in a protected environment which were Lactobacillus-free prior to inoculation. DNAs and RNAs extracted from the digesta of the chickens were used for PCR and reverse transcription-PCR, respectively, to demonstrate the presence and transcription of lgs genes in vivo. In both cases, only the strain-specific gene was transcribed. Both of the strains adhered to the crop epithelium, consistent with published data predicting that S-proteins of lactobacilli are adhesins. The results of this study provide a basis for the investigation of gene duplication and sequence variation as mechanisms by which bacterial strains of the same species can share the same habitat. PMID- 16269693 TI - Insights into Actinobacillus succinogenes fermentative metabolism in a chemically defined growth medium. AB - Chemically defined media allow for a variety of metabolic studies that are not possible with undefined media. A defined medium, AM3, was created to expand the experimental opportunities for investigating the fermentative metabolism of succinate-producing Actinobacillus succinogenes. AM3 is a phosphate-buffered medium containing vitamins, minerals, NH4Cl as the main nitrogen source, and glutamate, cysteine, and methionine as required amino acids. A. succinogenes growth trends and end product distributions in AM3 and rich medium fermentations were compared. The effects of NaHCO3 concentration in AM3 on end product distribution, growth rate, and metabolic rates were also examined. The A. succinogenes growth rate was 1.3 to 1.4 times higher at an NaHCO3 concentration of 25 mM than at any other NaHCO3 concentration, likely because both energy producing metabolic branches (i.e., the succinate-producing branch and the formate-, acetate-, and ethanol-producing branch) were functioning at relatively high rates in the presence of 25 mM bicarbonate. To improve the accuracy of the A. succinogenes metabolic map, the reasons for A. succinogenes glutamate auxotrophy were examined by enzyme assays and by testing the ability of glutamate precursors to support growth. Enzyme activities were detected for glutamate synthesis that required glutamine or alpha-ketoglutarate. The inability to synthesize alpha-ketoglutarate from glucose indicates that at least two tricarboxylic acid cycle-associated enzyme activities are absent in A. succinogenes. PMID- 16269694 TI - Ability of Shiga toxin-producing Escherichia coli and Salmonella spp. to survive in a desiccation model system and in dry foods. AB - In order to determine desiccation tolerances of bacterial strains, the survival of 58 diarrheagenic strains (18 salmonellae, 35 Shiga toxin-producing Escherichia coli [STEC], and 5 shigellae) and of 15 nonpathogenic E. coli strains was determined after drying at 35 degrees C for 24 h in paper disks. At an inoculum level of 10(7) CFU/disk, most of the salmonellae (14/18) and the STEC strains (31/35) survived with a population of 10(3) to 10(4) CFU/disk, whereas all of the shigellae (5/5) and the majority of the nonpathogenic E. coli strains (9/15) did not survive (the population was decreased to less than the detection limit of 10(2) CFU/disk). After 22 to 24 months of subsequent storage at 4 degrees C, all of the selected salmonellae (4/4) and most of the selected STEC strains (12/15) survived, keeping the original populations (10(3) to 10(4) CFU/disk). In contrast to the case for storage at 4 degrees C, all of 15 selected strains (5 strains each of Salmonella spp., STEC O157, and STEC O26) died after 35 to 70 days of storage at 25 degrees C and 35 degrees C. The survival rates of all of these 15 strains in paper disks after the 24 h of drying were substantially increased (10 to 79 times) by the presence of sucrose (12% to 36%). All of these 15 desiccated strains in paper disks survived after exposure to 70 degrees C for 5 h. The populations of these 15 strains inoculated in dried foods containing sucrose and/or fat (e.g., chocolate) were 100 times higher than those in the dried paper disks after drying for 24 h at 25 degrees C. PMID- 16269695 TI - Impact of different in vitro electron donor/acceptor conditions on potential chemolithoautotrophic communities from marine pelagic redoxclines. AB - Anaerobic or microaerophilic chemolithoautotrophic bacteria have been considered to be responsible for CO2 dark fixation in different pelagic redoxclines worldwide, but their involvement in redox processes is still not fully resolved. We investigated the impact of 17 different electron donor/acceptor combinations in water of pelagic redoxclines from the central Baltic Sea on the stimulation of bacterial CO2 dark fixation as well as on the development of chemolithoautotrophic populations. In situ, the highest CO2 dark fixation rates, ranging from 0.7 to 1.4 micromol liter(-1) day(-1), were measured directly below the redoxcline. In enrichment experiments, chemolithoautotrophic CO2 dark fixation was maximally stimulated by the addition of thiosulfate, reaching values of up to 9.7 micromol liter(-1) CO2 day(-1). Chemolithoautotrophic nitrate reduction proved to be an important process, with rates of up to 33.5 micromol liter(-1) NO3(-) day(-1). Reduction of Fe(III) or Mn(IV) was not detected; nevertheless, the presence of these potential electron acceptors influenced the development of stimulated microbial assemblages. Potential chemolithoautotrophic bacteria in the enrichment experiments were displayed on 16S ribosomal complementary DNA single-strand-conformation polymorphism fingerprints and identified by sequencing of excised bands. Sequences were closely related to chemolithoautotrophic Thiomicrospira psychrophila and Maorithyas hadalis gill symbiont (both Gammaproteobacteria) and to an uncultured nitrate-reducing Helicobacteraceae bacterium (Epsilonproteobacteria). Our data indicate that this Helicobacteraceae bacterium could be of general importance or even a key organism for autotrophic nitrate reduction in pelagic redoxclines. PMID- 16269696 TI - Bacteria associated with spores of the arbuscular mycorrhizal fungi Glomus geosporum and Glomus constrictum. AB - Spores of the arbuscular mycorrhizal fungi (AMF) Glomus geosporum and Glomus constrictum were harvested from single-spore-derived pot cultures with either Plantago lanceolata or Hieracium pilosella as host plants. PCR-denaturing gradient gel electrophoresis analysis revealed that the bacterial communities associated with the spores depended more on AMF than host plant identity. The composition of the bacterial populations linked to the spores could be predominantly influenced by a specific spore wall composition or AMF exudate rather than by specific root exudates. The majority of the bacterial sequences that were common to both G. geosporum and G. constrictum spores were affiliated with taxonomic groups known to degrade biopolymers (Cellvibrio, Chondromyces, Flexibacter, Lysobacter, and Pseudomonas). Scanning electron microscopy of G. geosporum spores revealed that these bacteria are possibly feeding on the outer hyaline spore layer. The process of maturation and eventual germination of AMF spores might then benefit from the activity of the surface microorganisms degrading the outer hyaline wall layer. PMID- 16269697 TI - Molecular epidemiology of antimicrobial-resistant commensal Escherichia coli strains in a cohort of newborn calves. AB - Pulsed-field gel electrophoresis (PFGE) was used to investigate the dissemination and diversity of ampicillin-resistant (Amp(r)) and nalidixic acid-resistant (Nal(r)) commensal Escherichia coli strains in a cohort of 48 newborn calves. Calves were sampled weekly from birth for up to 21 weeks and a single resistant isolate selected from positive samples for genotyping and further phenotypic characterization. The Amp(r) population showed the greatest diversity, with a total of 56 different genotype patterns identified, of which 5 predominated, while the Nal(r) population appeared to be largely clonal, with over 97% of isolates belonging to just two different PFGE patterns. Distinct temporal trends were identified in the distribution of several Amp(r) genotypes across the cohort, with certain patterns predominating at different points in the study. Cumulative recognition of new Amp(r) genotypes within the cohort was biphasic, with a turning point coinciding with the housing of the cohort midway through the study, suggesting that colonizing strains were from an environmental source on the farm. Multiply resistant isolates dominated the collection, with >95% of isolates showing resistance to at least two additional antimicrobials. Carriage of resistance to streptomycin, sulfamethoxazole, and tetracycline was the most common combination, found across several different genotypes, suggesting the possible spread of a common resistance element across multiple strains. The proportion of Amp(r) isolates carrying sulfamethoxazole resistance increased significantly over the study period (P < 0.05), coinciding with a decline in the most common genotype pattern. These data indicate that calves were colonized by a succession of multiply resistant strains, with a probable environmental source, that disseminated through the cohort over time. PMID- 16269699 TI - Intervening sequence acquired by lateral gene transfer in Tropheryma whipplei results in 23S rRNA fragmentation. AB - Completion of Tropheryma whipplei genome sequencing may provide insights into the evolution of the molecular mechanisms underlying the pathogenicity of this microorganism. The first postgenomic application was the successful design of a comprehensive culture medium that allows axenic growth of this bacterium, which is particularly recalcitrant to cultivation. This achievement in turn permitted analysis of T. whipplei RNA without contaminating eukaryotic nucleic acids. To obtain high-quality RNA, several extraction methods were compared, but under all conditions tested an atypical profile was observed. By using a Northern blot assay we demonstrated that an insertion sequence previously described in T. whipplei 23S rRNA is in fact an intervening sequence excised during maturation. This cleavage could involve an RNase III identified in the genome of this microorganism. Among the bacteria with a 23S rRNA insertion sequence, T. whipplei is the only gram-positive microorganism. We present phylogenetic evidence that this mobile genetic element was acquired by lateral gene transfer from another enteric bacterium. PMID- 16269698 TI - Identification of Shewanella baltica as the most important H2S-producing species during iced storage of Danish marine fish. AB - Shewanella putrefaciens has been considered the main spoilage bacteria of low temperature stored marine seafood. However, psychrotropic Shewanella have been reclassified during recent years, and the purpose of the present study was to determine whether any of the new Shewanella species are important in fish spoilage. More than 500 H2S-producing strains were isolated from iced stored marine fish (cod, plaice, and flounder) caught in the Baltic Sea during winter or summer time. All strains were identified as Shewanella species by phenotypic tests. Different Shewanella species were present on newly caught fish. During the warm summer months the mesophilic human pathogenic S. algae dominated the H2S producing bacterial population. After iced storage, a shift in the Shewanella species was found, and most of the H2S-producing strains were identified as S. baltica. The 16S rRNA gene sequence analysis confirmed the identification of these two major groups. Several isolates could only be identified to the genus Shewanella level and were separated into two subgroups with low (44%) and high (47%) G+C mol%. The low G+C% group was isolated during winter months, whereas the high G+C% group was isolated on fish caught during summer and only during the first few days of iced storage. Phenotypically, these strains were different from the type strains of S. putrefaciens, S. oneidensis, S. colwelliana, and S. affinis, but the high G+C% group clustered close to S. colwelliana by 16S rRNA gene sequence comparison. The low G+C% group may constitute a new species. S. baltica, and the low G+C% group of Shewanella spp. strains grew well in cod juice at 0 degrees C, but three high G+C Shewanella spp. were unable to grow at 0 degrees C. In conclusion, the spoilage reactions of iced Danish marine fish remain unchanged (i.e., trimethylamine-N-oxide reduction and H2S production); however, the main H2S-producing organism was identified as S. baltica. PMID- 16269700 TI - On-site DNA extraction and real-time PCR for detection of Phytophthora ramorum in the field. AB - Phytophthora ramorum is a recently described pathogen causing oak mortality (sudden oak death) in forests in coastal areas of California and southern Oregon and dieback and leaf blight in a range of tree, shrub, and herbaceous species in the United States and Europe. Due to the threat posed by this organism, stringent quarantine regulations are in place, which restrict the movement of a number of hosts. Fast and accurate diagnostic tests are required in order to characterize the distribution of P. ramorum, prevent its introduction into pathogen-free areas, and minimize its spread within affected areas. However, sending samples to a laboratory for testing can cause a substantial delay between sampling and diagnosis. A rapid and simple DNA extraction method was developed for use at the point of sampling and used to extract DNAs from symptomatic foliage and stems in the field. A sensitive and specific single-round real-time PCR (TaqMan) assay for P. ramorum was performed using a portable real-time PCR platform (Cepheid SmartCycler II), and a cost-effective method for stabilizing PCR reagents was developed to allow their storage and transportation at room temperature. To our knowledge, this is the first description of a method for DNA extraction and molecular testing for a plant pathogen carried out entirely in the field, independent of any laboratory facilities. PMID- 16269701 TI - Degradation of azo dyes by Trametes villosa laccase over long periods of oxidative conditions. AB - Trametes villosa laccase was used for direct azo dye degradation, and the reaction products that accumulated after 72 h of incubation were analyzed. Liquid chromatography-mass spectrometry (LC-MS) analysis showed the formation of phenolic compounds during the dye oxidation process as well as a large amount of polymerized products that retain azo group integrity. The amino-phenol reactions were also investigated by 13C-nuclear magnetic resonance and LC-MS analysis, and the polymerization character of laccase was shown. This study highlights the fact that laccases polymerize the reaction products obtained during long-term batch decolorization processes with azo dyes. These polymerized products provide unacceptable color levels in effluents, limiting the application of laccases as bioremediation agents. PMID- 16269702 TI - Impact of Bt corn on rhizospheric and soil eubacterial communities and on beneficial mycorrhizal symbiosis in experimental microcosms. AB - A polyphasic approach has been developed to gain knowledge of suitable key indicators for the evaluation of environmental impact of genetically modified Bt 11 and Bt 176 corn lines on soil ecosystems. We assessed the effects of Bt corn (which constitutively expresses the insecticidal toxin from Bacillus thuringiensis, encoded by the truncated Cry1Ab gene) and non-Bt corn plants and their residues on rhizospheric and bulk soil eubacterial communities by means of denaturing gradient gel electrophoresis analyses of 16S rRNA genes, on the nontarget mycorrhizal symbiont Glomus mosseae, and on soil respiration. Microcosm experiments showed differences in rhizospheric eubacterial communities associated with the three corn lines and a significantly lower level of mycorrhizal colonization in Bt 176 corn roots. In greenhouse experiments, differences between Bt and non-Bt corn plants were detected in rhizospheric eubacterial communities (both total and active), in culturable rhizospheric heterotrophic bacteria, and in mycorrhizal colonization. Plant residues of transgenic plants, plowed under at harvest and kept mixed with soil for up to 4 months, affected soil respiration, bacterial communities, and mycorrhizal establishment by indigenous endophytes. The multimodal approach utilized in our work may be applied in long-term field studies aimed at monitoring the real hazard of genetically modified crops and their residues on nontarget soil microbial communities. PMID- 16269703 TI - Detection of Salmonella enterica in naturally contaminated liquid eggs by loop mediated isothermal amplification, and characterization of Salmonella isolates. AB - Loop-mediated isothermal amplification (LAMP) assay was effective in detecting Salmonella enterica in naturally contaminated liquid egg samples. Salmonella was detected in 110 samples taken from four egg-breaking plants. The egg samples were pre-enriched in buffered peptone water (BPW) at 37 degrees C for 20 h. The selective enrichment was done in Rappaport-Vassiliadis or tetrathionate broth and plated onto xylose lysine deoxycholate agar and brilliant green agar, modified. In addition, the PCR assay was used to detect Salmonella after pre-enrichment in BPW at 37 degrees C for 20 h. The culture method and PCR assay were compared to the LAMP assay, which was also performed after pre-enrichment in BPW. PCR failed to detect Salmonella in 10% of 110 samples, whereas the culture method and LAMP assay successfully identified Salmonella in all samples. However, the LAMP assay was found to be much more rapid than the culture method and as sensitive in detecting Salmonella from liquid eggs. In all of the egg-breaking plants studied, Salmonella was isolated on most tested days. The positive samples showed that more than 75% of the Salmonella strains had identical genetic patterns when analyzed by pulsed-field gel electrophoresis. This suggests that the same Salmonella strains having survived long periods of time in the plants were contaminating the production line. The LAMP assay is rapid, specific, and sensitive for Salmonella detection in liquid eggs and is able to monitor Salmonella contamination in egg-handling plants more reliably. PMID- 16269704 TI - Cloning, biochemical properties, and distribution of mycobacterial haloalkane dehalogenases. AB - Haloalkane dehalogenases are enzymes that catalyze the cleavage of the carbon halogen bond by a hydrolytic mechanism. Genomes of Mycobacterium tuberculosis and M. bovis contain at least two open reading frames coding for the polypeptides showing a high sequence similarity with biochemically characterized haloalkane dehalogenases. We describe here the cloning of the haloalkane dehalogenase genes dmbA and dmbB from M. bovis 5033/66 and demonstrate the dehalogenase activity of their translation products. Both of these genes are widely distributed among species of the M. tuberculosis complex, including M. bovis, M. bovis BCG, M. africanum, M. caprae, M. microti, and M. pinnipedii, as shown by the PCR screening of 48 isolates from various hosts. DmbA and DmbB proteins were heterologously expressed in Escherichia coli and purified to homogeneity. The DmbB protein had to be expressed in a fusion with thioredoxin to obtain a soluble protein sample. The temperature optimum of DmbA and DmbB proteins determined with 1,2-dibromoethane is 45 degrees C. The melting temperature assessed by circular dichroism spectroscopy of DmbA is 47 degrees C and DmbB is 57 degrees C. The pH optimum of DmbA depends on composition of a buffer with maximal activity at 9.0. DmbB had a single pH optimum at pH 6.5. Mycobacteria are currently the only genus known to carry more than one haloalkane dehalogenase gene, although putative haloalkane dehalogenases can be inferred in more then 20 different bacterial species by comparative genomics. The evolution and distribution of haloalkane dehalogenases among mycobacteria is discussed. PMID- 16269705 TI - Abundance, activity, and community structure of pelagic methane-oxidizing bacteria in temperate lakes. AB - The abundance and activity of methane-oxidizing bacteria (MOB) in the water column were investigated in three lakes with different contents of nutrients and humic substances. The abundance of MOB was determined by analysis of group specific phospholipid fatty acids from type I and type II MOB, and in situ activity was measured with a 14CH4 transformation method. The fatty acid analyses indicated that type I MOB most similar to species of Methylomonas, Methylomicrobium, and Methylosarcina made a substantial contribution (up to 41%) to the total bacterial biomass, whereas fatty acids from type II MOB generally had very low concentrations. The MOB biomass and oxidation activity were positively correlated and were highest in the hypo- and metalimnion during summer stratification, whereas under ice during winter, maxima occurred close to the sediments. The methanotroph biomass-specific oxidation rate (V) ranged from 0.001 to 2.77 mg CH4-C mg(-1) C day(-1) and was positively correlated with methane concentration, suggesting that methane supply largely determined the activity and biomass distribution of MOB. Our results demonstrate that type I MOB often are a large component of pelagic bacterial communities in temperate lakes. They represent a potentially important pathway for reentry of carbon and energy into pelagic food webs that would otherwise be lost as evasion of CH4. PMID- 16269707 TI - Enhanced hydrogen production from formic acid by formate hydrogen lyase overexpressing Escherichia coli strains. AB - Genetic recombination of Escherichia coli in conjunction with process manipulation was employed to elevate the efficiency of hydrogen production in the resultant strain SR13 2 orders of magnitude above that of conventional methods. The formate hydrogen lyase (FHL)-overexpressing strain SR13 was constructed by combining FHL repressor (hycA) inactivation with FHL activator (fhlA) overexpression. Transcription of large-subunit formate dehydrogenase, fdhF, and large-subunit hydrogenase, hycE, in strain SR13 increased 6.5- and 7.0-fold, respectively, compared to the wild-type strain. On its own, this genetic modification effectively resulted in a 2.8-fold increase in hydrogen productivity of SR13 compared to the wild-type strain. Further enhancement of productivity was attained by using a novel method involving the induction of the FHL complex with high-cell-density filling of a reactor under anaerobic conditions. Continuous hydrogen production was achieved by maintaining the reactor concentration of the substrate (free formic acid) under 25 mM. An initial productivity of 23.6 g hydrogen h(-1) liter(-1) (300 liters h(-1) liter(-1) at 37 degrees C) was achieved using strain SR13 at a cell density of 93 g (dry weight) cells/liter. The hydrogen productivity reported in this work has great potential for practical application. PMID- 16269706 TI - Antimicrobial resistance and virulence genes of Escherichia coli isolates from swine in Ontario. AB - A total of 318 Escherichia coli isolates obtained from diarrheic and healthy pigs in Ontario from 2001 to 2003 were examined for their susceptibility to 19 antimicrobial agents. They were tested by PCR for the presence of resistance genes for tetracycline, streptomycin, sulfonamides, and apramycin and of 12 common virulence genes of porcine E. coli. Antimicrobial resistance frequency among E. coli isolates from swine in Ontario was moderate in comparison with other countries and was higher in isolates from pigs with diarrhea than in isolates from healthy finisher pigs. Resistance profiles suggest that cephamycinases may be produced by > or = 8% of enterotoxigenic E. coli (ETEC). Resistance to quinolones was detected only in enterotoxigenic E. coli (< or = 3%). The presence of sul3 was demonstrated for the first time in Canada in porcine E. coli isolates. Associations were observed among tetA, sul1, aadA, and aac(3)IV and among tetB, sul2, and strA/strB, with a strong negative association between tetA and tetB. The paa and sepA genes were detected in 92% of porcine ETEC, and strong statistical associations due to colocation on a large plasmid were observed between tetA, estA, paa, and sepA. Due at least in part to gene linkages, the distribution of resistance genes was very different between ETEC isolates and other porcine E. coli isolates. This demonstrates that antimicrobial resistance epidemiology differs significantly between pathogenic and commensal E. coli isolates. These results may have important implications with regards to the spread and persistence of resistance and virulence genes in bacterial populations and to the prudent use of antimicrobial agents. PMID- 16269709 TI - Effect of different carbon sources on community composition of bacterial enrichments from soil. AB - Soil is a highly heterogeneous matrix, which can contain thousands of different bacterial species per gram. Only a small component of this diversity (maybe <1%) is commonly captured using standard isolation techniques, although indications are that a larger proportion of the soil community is in fact culturable. Better isolation techniques yielding greater bacterial diversity would be of benefit for understanding the metabolic activity and capability of many soil microorganisms. We studied the response of soil bacterial communities to carbon source enrichment in small matrices by means of terminal restriction fragment length polymorphism (TRFLP) analysis. The community composition of replicate enrichments from soil displayed high variability, likely attributable to soil heterogeneity. An analysis of TRFLP data indicated that enrichment on structurally similar carbon sources selected for similar bacterial communities. The same analysis indicated that communities first enriched on glucose or benzoate and subsequently transferred into medium containing an alternate carbon source retained a distinct community signature induced by the carbon source used in the primary enrichment. Enrichment on leucine presented a selective challenge that was able to override the imprint left by primary enrichment on acetate. In a time series experiment community change was most rapid 18 hours after inoculation, corresponding to exponential growth. Community composition did not stabilize even 4 days after secondary enrichment. Four different soil types were enriched on four different carbon sources. TRFLP analysis indicated that in three out of four cases communities enriched on the same carbon source were more similar regardless of which soil type was used. Conversely, the garden soil samples yielded similar enrichment communities regardless of the enrichment carbon source. Our results indicate that in order to maximize the diversity of bacteria recovered from the environment, multiple enrichments should be performed using a chemically diverse set of carbon sources. PMID- 16269708 TI - Expression of rumen microbial fibrolytic enzyme genes in probiotic Lactobacillus reuteri. AB - This study was aimed at evaluating the cloning and expression of three rumen microbial fibrolytic enzyme genes in a strain of Lactobacillus reuteri and investigating the probiotic characteristics of these genetically modified lactobacilli. The Neocallimastix patriciarum xylanase gene xynCDBFV, the Fibrobacter succinogenes beta-glucanase (1,3-1,4-beta-D-glucan 4-glucanohydrolase [EC 3.2.1.73]) gene, and the Piromyces rhizinflata cellulase gene eglA were cloned in a strain of L. reuteri isolated from the gastrointestinal tract of broilers. The enzymes were expressed and secreted under the control of the Lactococcus lactis lacA promoter and its secretion signal. The L. reuteri transformed strains not only acquired the capacity to break down soluble carboxymethyl cellulose, beta-glucan, or xylan but also showed high adhesion efficiency to mucin and mucus and resistance to bile salt and acid. PMID- 16269710 TI - Links between plant and rhizoplane bacterial communities in grassland soils, characterized using molecular techniques. AB - Molecular analysis of grassland rhizosphere soil has demonstrated complex and diverse bacterial communities, with resultant difficulties in detecting links between plant and bacterial communities. These studies have, however, analyzed "bulk" rhizosphere soil, rather than rhizoplane communities, which interact most closely with plants through utilization of root exudates. The aim of this study was to test the hypothesis that plant species was a major driver for bacterial rhizoplane community composition on individual plant roots. DNA extracted from individual roots was used to determine plant identity, by analysis of the plastid tRNA leucine (trnL) UAA gene intron, and plant-related bacterial communities. Bacterial communities were characterized by analysis of PCR-amplified 16S rRNA genes using two fingerprinting methods: terminal restriction fragment length polymorphisms (T-RFLP) and denaturing gradient gel electrophoresis (DGGE). Links between plant and bacterial rhizoplane communities could not be detected by visual examination of T-RFLP patterns or DGGE banding profiles. Statistical analysis of fingerprint patterns did not reveal a relationship between bacterial community composition and plant species but did demonstrate an influence of plant community composition. The data also indicated that topography and other, uncharacterized, environmental factors are important in driving bacterial community composition in grassland soils. T-RFLP had greater potential resolving power than DGGE, but findings from the two methods were not significantly different. PMID- 16269711 TI - Detection of infectious Cryptosporidium oocysts by cell culture immunofluorescence assay: applicability to environmental samples. AB - In the past few years many waterborne outbreaks related to Cryptosporidium have been described. Current methods for detection of Cryptosporidium in water for the most part rely on viability assays which are not informative concerning the infectivity of oocysts. However, for estimation of the risk of infection with Cryptosporidium this information is required. For environmental samples the oocyst counts are often low, and the oocysts have been exposed to unfavorable conditions. Therefore, determination of the infectivity of environmental oocysts requires an assay with a high level of sensitivity. We evaluated the applicability of in vitro cell culture immunofluorescence assays with HCT-8 and Caco-2 cells for determination of oocyst infectivity in naturally contaminated water samples. Cell culture assays were compared with other viability and infectivity assays. Experiments with Cryptosporidium oocysts from different sources revealed that there was considerable variability in infectivity, which was illustrated by variable 50% infective doses, which ranged from 40 to 614 oocysts, and the results indicated that not only relatively large numbers of fresh oocysts but also aged oocysts produced infection in cell cultures. Fifteen Dutch surface water samples were tested, and the cell culture immunofluorescence assays were not capable of determining the infectivity for the low numbers of naturally occurring Cryptosporidium oocysts present in the samples. A comparison with other viability assays, such as the vital dye exclusion assay, demonstrated that surrogate methods overestimate the number of infectious oocysts and therefore the risk of infection with Cryptosporidium. For accurate risk assessment, further improvement of the method for detection of Cryptosporidium in water is needed. PMID- 16269712 TI - Growth and grazing mortality rates of phylogenetic groups of bacterioplankton in coastal marine environments. AB - Dilution culture experiments were conducted in western North Pacific coastal regions to determine growth and grazing mortality rates of bacterial phylogenetic groups (alpha-, beta-, and gamma-proteobacteria and the Cytophaga-Flavobacter cluster) detected by fluorescent in situ hybridization. Growth rates varied greatly (1.2- to 4.0-fold) among different groups, and they were related to environmental variables (chlorophyll a concentrations and temperature) in a group specific fashion. Growth rates of alpha-proteobacteria, the most abundant group in all the samples examined, were generally lower than those of less abundant groups, including the Cytophaga-Flavobacter cluster and gamma-proteobacteria. Grazing mortality rates and mean cell volumes varied little among different groups. These results provide insights into factors that affect distributions of different groups, but growth and grazing mortality alone did not fully explain bacterial community compositions at a broad phylogenetic level. PMID- 16269713 TI - Polyphenol oxidase activity expression in Ralstonia solanacearum. AB - Sequencing of the genome of Ralstonia solanacearum revealed several genes that putatively code for polyphenol oxidases (PPOs). To study the actual expression of these genes, we looked for and detected all kinds of PPO activities, including laccase, cresolase, and catechol oxidase activities, in cellular extracts of this microorganism. The conditions for the PPO assays were optimized for the phenolic substrate, pH, and sodium dodecyl sulfate concentration used. It was demonstrated that three different PPOs are expressed. The genes coding for the enzymes were unambiguously correlated with the enzymatic activities detected by generation of null mutations in the genes by using insertional mutagenesis with a suicide plasmid and estimating the changes in the levels of enzymatic activities compared to the levels in the wild-type strain. The protein encoded by the RSp1530 locus is a multicopper protein with laccase activity. Two other genes, RSc0337 and RSc1501, code for nonblue copper proteins exhibiting homology to tyrosinases. The product of RSc0337 has strong tyrosine hydroxylase activity, and it has been shown that this enzyme is involved in melanin synthesis by R. solanacearum. The product of the RSc1501 gene is an enzyme that shows a clear preference for oxidation of o-diphenols. Preliminary characterization of the mutants obtained indicated that PPOs expressed by R. solanacearum may participate in resistance to phenolic compounds since the mutants exhibited higher sensitivity to L-tyrosine than the wild-type strain. These results suggest a possible role in the pathogenic process to avoid plant resistance mechanisms involving the participation of phenolic compounds. PMID- 16269714 TI - Escherichia coli O157:H7 in environments of culture-positive cattle. AB - Outbreaks of Escherichia coli O157:H7 disease associated with animal exhibits have been reported with increasing frequency. Transmission can occur through contact with contaminated haircoats, bedding, farm structures, or water. We investigated the distribution and survival of E. coli O157:H7 in the immediate environments of individually housed, experimentally inoculated cattle by systematically culturing feed, bedding, water, haircoat, and feed bunk walls for E. coli O157:H7 for 3 months. Cedar chip bedding was the most frequently culture positive environmental sample tested (27/96 or 28.15%). Among these, 12 (44.0%) of positive bedding samples were collected when the penned animal was fecal culture negative. Survival of E. coli O157:H7 in experimentally inoculated cedar chip bedding and in grass hay feed was determined at different temperatures. Survival was longest in feed at room temperature (60 days), but bacterial counts decreased over time. The possibility that urine plays a role in the environmental survival of E. coli O157:H7 was investigated. Cedar chip bedding moistened with sterile water or bovine urine was inoculated with E. coli O157:H7. Bedding moistened with urine supported growth of E. coli O157:H7, whereas inoculated bedding moistened with only water yielded decreasing numbers of bacteria over time. The findings that environmental samples were frequently positive for E. coli O157:H7 at times when animals were culture negative and that urine provided a substrate for E. coli O157:H7 growth have implications for understanding the on farm ecology of this pathogen and for the safety of ruminant animal exhibits, particularly petting zoos and farms where children may enter animal pens. PMID- 16269715 TI - Rapid identification and enumeration of Saccharomyces cerevisiae cells in wine by real-time PCR. AB - Despite the beneficial role of Saccharomyces cerevisiae in the food industry for food and beverage production, it is able to cause spoilage in wines. We have developed a real-time PCR method to directly detect and quantify this yeast species in wine samples to provide winemakers with a rapid and sensitive method to detect and prevent wine spoilage. Specific primers were designed for S. cerevisiae using the sequence information obtained from a cloned random amplified polymorphic DNA band that differentiated S. cerevisiae from its sibling species Saccharomyces bayanus, Saccharomyces pastorianus, and Saccharomyces paradoxus. The specificity of the primers was demonstrated for typical wine spoilage yeast species. The method was useful for estimating the level of S. cerevisiae directly in sweet wines and red wines without preenrichment when yeast is present in concentrations as low as 3.8 and 5 CFU per ml. This detection limit is in the same order as that obtained from glucose-peptone-yeast growth medium (GPY). Moreover, it was possible to quantify S. cerevisiae in artificially contaminated samples accurately. Limits for accurate quantification in wine were established, from 3.8 x 10(5) to 3.8 CFU/ml in sweet wine and from 5 x 10(6) to 50 CFU/ml in red wine. PMID- 16269716 TI - Monitoring stress-related genes during the process of biomass propagation of Saccharomyces cerevisiae strains used for wine making. AB - Physiological capabilities and fermentation performance of Saccharomyces cerevisiae strains to be employed during industrial wine fermentations are critical for the quality of the final product. During the process of biomass propagation, yeast cells are dynamically exposed to a mixed and interrelated group of known stresses such as osmotic, oxidative, thermic, and/or starvation. These stressing conditions can dramatically affect the parameters of the fermentation process and the technological abilities of the yeast, e.g., the biomass yield and its fermentative capacity. Although a good knowledge exists of the behavior of S. cerevisiae under laboratory conditions, insufficient knowledge is available about yeast stress responses under the specific media and growth conditions during industrial processes. We performed growth experiments using bench-top fermentors and employed a molecular marker approach (changes in expression levels of five stress-related genes) to investigate how the cells respond to environmental changes during the process of yeast biomass production. The data show that in addition to the general stress response pathway, using the HSP12 gene as a marker, other specific stress response pathways were induced, as indicated by the changes detected in the mRNA levels of two stress-related genes, GPD1 and TRX2. These results suggest that the cells were affected by osmotic and oxidative stresses, demonstrating that these are the major causes of the stress response throughout the process of wine yeast biomass production. PMID- 16269717 TI - Enteroviruses and bacteriophages in bathing waters. AB - A new procedure for detecting and counting enteroviruses based on the VIRADEN method applied to 10 liters of seawater was examined. It improved the efficiency of detection by taking into account both the number of positive isolations and numbers found with traditional methods. It was then used to quantify viruses in bathing waters. A number of bacterial indicators and bacteriophages were also tested. Cultivable enteroviruses were detected in 55% of the samples, most of which complied with bacteriological criteria. In contrast, viral genomes were only detected in 20% of the samples by reverse transcription-PCR. Somatic coliphages outnumbered all other indicators. F-specific RNA phages were detected in only 15% of the samples, whereas phages infecting Bacteroides thetaiotaomicron were detected in 70% of samples. A numerical relationship between the numbers of enteroviruses and the numbers of enterococci and somatic coliphages was observed. In situ inactivation experiments showed that viruses persisted significantly longer than the bacterial indicators. Only somatic coliphages and bacteriophages infecting Bacteroides persisted longer than the viruses. These results explain the numbers of enteroviruses and indicators in bathing waters attending the numbers usually found in sewage in the area. Somatic coliphages show a very good potential to predict the risk of viruses being present in bathing waters. PMID- 16269718 TI - Identification and characterization of a novel biotin biosynthesis gene in Saccharomyces cerevisiae. AB - Yeast Saccharomyces cerevisiae cells generally cannot synthesize biotin, a vitamin required for many carboxylation reactions. Although sake yeasts, which are used for Japanese sake brewing, are classified as S. cerevisiae, they do not require biotin for their growth. In this study, we identified a novel open reading frame (ORF) in the genome of one strain of sake yeast that we speculated to be involved in biotin synthesis. Homologs of this gene are widely distributed in the genomes of sake yeasts. However, they are not found in many laboratory strains and strains used for wine making and beer brewing. This ORF was named BIO6 because it has 52% identity with BIO3, a biotin biosynthesis gene of a laboratory strain. Further research showed that yeasts without the BIO6 gene are auxotrophic for biotin, whereas yeasts holding the BIO6 gene are prototrophic for biotin. The BIO6 gene was disrupted in strain A364A, which is a laboratory strain with one copy of the BIO6 gene. Although strain A364A is prototrophic for biotin, a BIO6 disrupted mutant was found to be auxotrophic for biotin. The BIO6 disruptant was able to grow in biotin-deficient medium supplemented with 7-keto-8 amino-pelargonic acid (KAPA), while the bio3 disruptant was not able to grow in this medium. These results suggest that Bio6p acts in an unknown step of biotin synthesis before KAPA synthesis. Furthermore, we demonstrated that expression of the BIO6 gene, like that of other biotin synthesis genes, was upregulated by depletion of biotin. We conclude that the BIO6 gene is a novel biotin biosynthesis gene of S. cerevisiae. PMID- 16269720 TI - Common, but complex, mode of resistance of Plutella xylostella to Bacillus thuringiensis toxins Cry1Ab and Cry1Ac. AB - A field collected population of Plutella xylostella (SERD4) was selected in the laboratory with Bacillus thuringiensis endotoxins Cry1Ac (Cry1Ac-SEL) and Cry1Ab (Cry1Ab-SEL). Both subpopulations showed similar phenotypes: high resistance to the Cry1A toxins and little cross-resistance to Cry1Ca or Cry1D. A previous analysis of the Cry1Ac-SEL showed incompletely dominant resistance to Cry1Ac with more than one factor, at least one of which was sex influenced. In the present study reciprocal mass crosses between Cry1Ab-SEL and a laboratory susceptible population (ROTH) provided evidence that Cry1Ab resistance was also inherited as incompletely dominant trait with more than one factor, and at least one of the factors was sex influenced. Analysis of single pair mating indicated that Cry1Ab SEL was still heterogeneous for Cry1Ab resistance genes, showing genes with different degrees of dominance. Binding studies showed a large reduction of specific binding of Cry1Ab and Cry1Ac to midgut membrane vesicles of the Cry1Ab SEL subpopulation. Cry1Ab-SEL was found to be more susceptible to trypsin activated Cry1Ab toxin than protoxin, although no defect in toxin activation was found. Present and previous results indicate a common basis of resistance to both Cry1Ab and Cry1Ac in selected subpopulations and suggest that a similar set of resistance genes are responsible for resistance to Cry1Ab and Cry1Ac and are selected whichever toxin was used. The possibility of an incompletely dominant trait of resistant to these toxins should be taken into account when considering refuge resistance management strategies. PMID- 16269719 TI - A propionate-inducible expression system for enteric bacteria. AB - A series of new expression vectors (pPro) have been constructed for the regulated expression of genes in Escherichia coli. The pPro vectors contain the prpBCDE promoter (P(prpB)) responsible for expression of the propionate catabolic genes (prpBCDE) and prpR encoding the positive regulator of this promoter. The efficiency and regulatory properties of the prpR-P(prpB) system were measured by placing the gene encoding the green fluorescent protein (gfp) under the control of the inducible P(prpB) of E. coli. This system provides homogenous expression in individual cells, highly regulatable expression over a wide range of propionate concentrations, and strong expression (maximal 1,500-fold induction) at high propionate concentrations. Since the prpBCDE promoter has CAP-dependent activation, the prpR-P(prpB) system exhibited negligible basal expression by addition of glucose to the medium. PMID- 16269721 TI - Potential for quantifying expression of the Geobacteraceae citrate synthase gene to assess the activity of Geobacteraceae in the subsurface and on current harvesting electrodes. AB - The Geobacteraceae citrate synthase is phylogenetically distinct from those of other prokaryotes and is a key enzyme in the central metabolism of Geobacteraceae. Therefore, the potential for using levels of citrate synthase mRNA to estimate rates of Geobacter metabolism was evaluated in pure culture studies and in four different Geobacteraceae-dominated environments. Quantitative reverse transcription-PCR studies with mRNA extracted from cultures of Geobacter sulfurreducens grown in chemostats with Fe(III) as the electron acceptor or in batch with electrodes as the electron acceptor indicated that transcript levels of the citrate synthase gene, gltA, increased with increased rates of growth/Fe(III) reduction or current production, whereas the expression of the constitutively expressed housekeeping genes recA, rpoD, and proC remained relatively constant. Analysis of mRNA extracted from groundwater collected from a U(VI)-contaminated site undergoing in situ uranium bioremediation revealed a remarkable correspondence between acetate levels in the groundwater and levels of transcripts of gltA. The expression of gltA was also significantly greater in RNA extracted from groundwater beneath a highway runoff recharge pool that was exposed to calcium magnesium acetate in June, when acetate concentrations were high, than in October, when the levels had significantly decreased. It was also possible to detect gltA transcripts on current-harvesting anodes deployed in freshwater sediments. These results suggest that it is possible to monitor the in situ metabolic rate of Geobacteraceae by tracking the expression of the citrate synthase gene. PMID- 16269722 TI - Development of a rapid method for simultaneous recovery of diverse microbes in drinking water by ultrafiltration with sodium polyphosphate and surfactants. AB - The ability to simultaneously concentrate diverse microbes is an important consideration for sample collection methods that are used for emergency response and environmental monitoring when drinking water may be contaminated with an array of unknown microbes. This study focused on developing a concentration method using ultrafilters and different combinations of a chemical dispersant (sodium polyphosphate [NaPP]) and surfactants. Tap water samples were seeded with bacteriophage MS2, Escherichia coli, Enterococcus faecalis, Cryptosporidium parvum, 4.5-microm microspheres, Salmonella enterica serovar Typhimurium, Bacillus globigii endospores, and echovirus 1. Ten-liter tap water samples were concentrated to approximately 250 ml in 12 to 42 min, depending on the experimental condition. Initial experiments indicated that pretreating filters with fetal bovine serum or NaPP resulted in an increase in microbe recovery. The addition of NaPP to the tap water samples resulted in significantly higher microbe and microsphere recovery efficiencies. Backflushing of the ultrafilter was found to significantly improve recovery efficiencies. The effectiveness of backflushing was improved further with the addition of Tween 80 to the backflush solution. The ultrafiltration method developed in this study, incorporating the use of NaPP pretreatment and surfactant solution backflushing, was found to recover MS2, C. parvum, microspheres, and several bacterial species with mean recovery efficiencies of 70 to 93%. The mean recovery efficiency for echovirus 1 (49%) was the lowest of the microbes studied for this method. This research demonstrates that ultrafiltration can be effective for recovering diverse microbes simultaneously in tap water and that chemical dispersants and surfactants can be beneficial for improving microbial recovery using this technique. PMID- 16269723 TI - Bacterial populations active in metabolism of C1 compounds in the sediment of Lake Washington, a freshwater lake. AB - Active members of the bacterial community in the sediment of Lake Washington, with special emphasis on C1 utilizers, were identified by employing two complementary culture-independent approaches: reverse transcription of environmental mRNA and 16S rRNA combined with PCR (RT-PCR) and stable-isotope probing (SIP) of DNA with the 13C-labeled C1 substrates methanol, methylamine, formaldehyde, and formate. Analysis of RT-PCR-amplified fragments of 16S rRNA encoding genes revealed that gammaproteobacterial methanotrophs belonging to Methylobacter and Methylomonas dominate the active methylotroph population, while only one other known methylotrophic lineage, Methylophilaceae, was detected via this approach. Analysis of RT-PCR-amplified functional genes, pmoA and fae, allowed detection of alphaproteobacterial (Methylosinus) and gammaproteobacterial (Methylobacter, Methylomonas, and Methylomicrobium) methanotrophs, methylotrophs of the genus Methylobacterium, and yet-unidentified proteobacteria. SIP experiments allowed detection of a broad variety of groups actively metabolizing C1 compounds. Comparisons between 16S rRNA gene pools amplified from [13C]DNA and from [12C]DNA revealed that the proportion of Methylophilus-related sequences increased in the presence of [13C]methanol, [13C]methylamine, and [13C]formaldehyde; Novosphingobium-related sequences were enriched in the presence of [13C]methanol; Gemmatimonadaceae-related sequences were enriched in the presence of [13C]formaldehyde and [13C]formate; and Xanthomonadaceae-related sequences were enriched in the presence of [13C]formate. Analysis of fae genes amplified from [13C]DNAs isolated from different microcosms revealed specific shifts in populations in response to a specific C1 compound: Methylosinus sequences dominated the [13C]methanol microcosm pool, and beta- and gammaproteobacterial sequences dominated the [13C]methylamine microcosm pool. The [13C]formaldehyde microcosm was dominated by betaproteobacterial sequences and by sequences of a nonaffiliated group, while the [13C]formate microcosm was dominated by alpha- and betaproteobacterial sequences. Overall, these data point toward the presence of a diverse population of active methylotrophs in Lake Washington sediments and toward the existence of yet-uncultivated organisms. PMID- 16269724 TI - Reciprocal regulation of pyoluteorin production with membrane transporter gene expression in Pseudomonas fluorescens Pf-5. AB - Pyoluteorin is a chlorinated polyketide antibiotic secreted by the rhizosphere bacterium Pseudomonas fluorescens Pf-5. Genes encoding enzymes and transcriptional regulators involved in pyoluteorin production are clustered in the genome of Pf-5. Sequence analysis of genes adjacent to the known pyoluteorin biosynthetic gene cluster revealed the presence of an ABC transporter system. We disrupted two putative ABC transporter genes by inserting transcriptional fusions to an ice nucleation reporter gene. Mutations in pltI and pltJ, which are predicted to encode a membrane fusion protein and an ATP-binding cassette of the ABC transporter, respectively, greatly reduced pyoluteorin production by Pf-5. During the transition from exponential growth to stationary phase, populations of a pltI mutant were lower than those of a pltI+ strain in a culture medium containing pyoluteorin, suggesting a role for the transport system in efflux and the resistance of Pf-5 to the antibiotic. Although pltI or pltJ mutant strains displayed low pyoluteorin production, they did not accumulate proportionately more of the antibiotic intracellularly, indicating that pltI and pltJ do not encode an exclusive exporter for pyoluteorin. Transcription of the putative pyoluteorin efflux genes pltI and pltJ was enhanced by exogenous pyoluteorin. These new observations parallel an earlier finding that pyoluteorin enhances the transcription of pyoluteorin biosynthesis genes and pyoluteorin production in Pf 5. This report provides evidence of a coordination of pyoluteorin production and the transcription of genes encoding a linked transport apparatus, wherein each requires the other for optimal expression. PMID- 16269725 TI - Efficient procedure for purification of obligate intracellular Wolbachia pipientis and representative amplification of its genome by multiple-displacement amplification. AB - Bacteria belonging to the genus Wolbachia are obligatory microendocytobionts that infect a variety of arthropods and a majority of filarial nematode species, where they induce reproductive alterations or establish a mutualistic symbiosis. Although two whole genome sequences of Wolbachia pipientis, for strain wMel from Drosophila melanogaster and strain wBm from Brugia malayi, have been fully completed and six other genome sequencing projects are ongoing (http://www.genomesonline.org/index.cgi?want=Prokaryotic+Ongoin), genetic analyses of these bacteria are still scarce, mainly due to the inability to cultivate them outside of eukaryotic cells. Usually, a large amount of host tissue (a thousand individuals, or about 10 g) is required in order to purify Wolbachia and extract its DNA, which is often recovered in small amounts and contaminated by host cell DNA, thus hindering genomic studies. In this report, we describe an efficient and reliable procedure to representatively amplify the Wolbachia genome by multiple-displacement amplification from limited infected host tissue (0.2 g or 2 x 10(7) cells). We obtained sufficient amounts (8 to 10 microg) of DNA of suitable quality for genomic studies, and we demonstrated that the amplified DNA contained all of the Wolbachia loci targeted. In addition, our data indicated that the genome of strain wRi, an obligatory endosymbiont of Drosophila simulans, shares a similar overall architecture with its relative strain wMel. PMID- 16269726 TI - Photodynamic inactivation of Bacillus spores, mediated by phenothiazinium dyes. AB - Spore formation is a sophisticated mechanism by which some bacteria survive conditions of stress and starvation by producing a multilayered protective capsule enclosing their condensed DNA. Spores are highly resistant to damage by heat, radiation, and commonly employed antibacterial agents. Previously, spores have also been shown to be resistant to photodynamic inactivation using dyes and light that easily destroy the corresponding vegetative bacteria. We have discovered that Bacillus spores are susceptible to photoinactivation by phenothiazinium dyes and low doses of red light. Dimethylmethylene blue, methylene blue, new methylene blue, and toluidine blue O are all effective, while alternative photosensitizers such as Rose Bengal, polylysine chlorin(e6) conjugate, a tricationic porphyrin, and a benzoporphyrin derivative, which easily kill vegetative cells, are ineffective. Spores of Bacillus cereus and B. thuringiensis are most susceptible, B. subtilis and B. atrophaeus are also killed, and B. megaterium is resistant. Photoinactivation is most effective when excess dye is washed from the spores, showing that the dye binds to the spores and that excess dye in solution can quench light delivery. The relatively mild conditions needed for spore killing could have applications for treating wounds contaminated by anthrax spores, for which conventional sporicides would have unacceptable tissue toxicity. PMID- 16269727 TI - Development and application of real-time PCR assays for quantification of genes encoding tetracycline resistance. AB - We report here the development, validation, and use of three real-time PCR assays to quantify the abundance of the following three groups of tetracycline resistance genes: tet(A) and tet(C); tet(G); and tet genes encoding ribosomal protection proteins, including tet(M), tet(O), tetB(P), tet(Q), tet(S), tet(T), and tet(W). The assays were validated using known numbers of sample-derived tet gene templates added to microbiome DNA. These assays are both precise and accurate over at least 6 log tet gene copies. New tet gene variants were also identified from cloned tet amplicons as part of this study. The utility of these real-time PCR assays was demonstrated by quantifying the three tet gene groups present in bovine and swine manures, composts of swine manure, lagoons of hog house effluent, and samples from an Ekokan upflow biofilter system treating hog house effluent. The bovine manures were found to contain fewer copies of all three groups of tet genes than the swine manures. The composts of swine manures had substantially reduced tet gene abundance (up to 6 log), while lagoon storage or the upflow biofilter had little effect on tet gene abundance. These results suggest that the method of manure storage and treatment may have a substantial impact on the persistence and dissemination of tet genes in agricultural environments. These real-time PCR assays provide rapid, quantitative, cultivation independent measurements of 10 major classes of tet genes, which should be useful for ecological studies of antibiotic resistance. PMID- 16269729 TI - Iron uptake mechanisms in the fish pathogen Tenacibaculum maritimum. AB - We present here the first evidence of the presence of iron uptake mechanisms in the bacterial fish pathogen Tenacibaculum maritimum. Representative strains of this species, with different serotypes and origins, were examined. All of them were able to grow in the presence of the chelating agent ethylenediamine-di-(o hydroxyphenyl acetic acid) (EDDHA) and also produced siderophores. Cross-feeding assays suggest that the siderophores produced are closely related. In addition, all T. maritimum strains utilized transferrin, hemin, hemoglobin, and ferric ammonic citrate as iron sources when added to iron-deficient media. Whole cells of all T. maritimum strains, grown under iron-supplemented or iron-restricted conditions, were able to bind hemin, indicating the existence of constitutive binding components located at the T. maritimum cell surface. This was confirmed by the observation that isolated total and outer membrane proteins from all of the strains, regardless of the iron levels of the media, were able to bind hemin, with the outer membranes showing the strongest binding. Proteinase K treatment of whole cells did not affect the hemin binding, indicating that, in addition to proteins, some protease-resistant components could also bind hemin. At least three outer membrane proteins were induced in iron-limiting conditions, and all strains, regardless of their serotype, showed a similar pattern of induced proteins. The results of the present study suggest that T. maritimum possesses at least two different systems of iron acquisition: one involving the synthesis of siderophores and another that allows the utilization of heme groups as iron sources by direct binding. PMID- 16269728 TI - Identifying components of the NF-kappaB pathway in the beneficial Euprymna scolopes-Vibrio fischeri light organ symbiosis. AB - The Toll/NF-kappaB pathway is a common, evolutionarily conserved innate immune pathway that modulates the responses of animal cells to microbe-associated molecular patterns (MAMPs). Because MAMPs have been implicated as critical elements in the signaling of symbiont-induced development, an expressed sequence tag library from the juvenile light organ of Euprymna scolopes was used to identify members of the Toll/NF-kappaB pathway. Full-length transcripts were identified by using 5' and 3' RACE PCR. Seven transcripts critical for MAMP induced triggering of the Toll/NF-kappaB phosphorylation cascade have been identified, including receptors, signal transducers, and a transcription factor. Further investigations should elucidate the role of the Toll/NF-kappaB pathway in the initiation of the beneficial symbiosis between E. scolopes and Vibrio fischeri. PMID- 16269730 TI - Putative transposases conserved in Exiguobacterium isolates from ancient Siberian permafrost and from contemporary surface habitats. AB - Gram-positive bacteria of the genus Exiguobacterium have been repeatedly isolated from Siberian permafrost ranging in age from 20,000 to 2 to 3 million years and have been sporadically recovered from markedly diverse habitats, including microbial mats in Lake Fryxell (Antarctic), surface water, and food-processing environments. However, there is currently no information on genomic diversity of this microorganism or on the physiological strategies that have allowed its survival under prolonged freezing in the permafrost. Analysis of the genome sequence of the most ancient available Exiguobacterium isolate (Exiguobacterium sp. strain 255-15, from 2 to 3 million-year-old Siberian permafrost) revealed numerous putative transposase sequences, primarily of the IS200/IS605, IS30, and IS3 families, with four transposase families identified. Several of the transposase genes appeared to be part of insertion sequences. Southern blots with different transposase probes yielded high-resolution genomic fingerprints which differentiated the different permafrost isolates from each other and from the Exiguobacterium spp. type strains which have been derived from diverse surface habitats. Each of the Exiguobacterium sp. strain 255-15 transposases that were used as probes had highly conserved homologs in the genome of other Exiguobacterium strains, both from permafrost and from modern sites. These findings suggest that, prior to their entrapment in permafrost, Exiguobacterium isolates had acquired transposases and that conserved transposases are present in Exiguobacterium spp., which now can be isolated from various modern surface habitats. PMID- 16269731 TI - Isolation of Mycobacterium avium subsp. paratuberculosis from free-ranging birds and mammals on livestock premises. AB - Surveys for Mycobacterium avium subsp. paratuberculosis infection in free-ranging mammals and birds were conducted on nine dairy and beef cattle farms in Wisconsin and Georgia. Specimens were collected from 774 animals representing 25 mammalian and 22 avian species. Specimens of ileum, liver, intestinal lymph nodes, and feces were harvested from the larger mammals; a liver specimen and the gastrointestinal tract were harvested from birds and small mammals. Cultures were performed by using radiometric culture and acid-fast isolates were identified by 16S/IS900/IS1311 PCR and mycobactin dependency characteristics. M. avium subsp. paratuberculosis was cultured from tissues and feces from 39 samples from 30 animals representing nine mammalian and three avian species. The prevalence of infected wild animals by premises ranged from 2.7 to 8.3% in Wisconsin and from 0 to 6.0% in Georgia. Shedding was documented in seven (0.9%) animals: three raccoons, two armadillos, one opossum, and one feral cat. The use of two highly polymorphic short sequence repeat loci for analysis of 29 of the 39 strains identified 10 alleles. One allelic pattern broadly shared in domestic ruminants ("7,5") appeared in approximately one-third of the wildlife M. avium subsp. paratuberculosis isolates studied. Given the few cases of shedding by free ranging animals compared to the volume of contaminated manure produced by infected domestic ruminant livestock, contamination of the farm environment by infected wildlife was negligible. Wildlife may, however, have epidemiological significance for farms where M. avium subsp. paratuberculosis recently has been eliminated or on farms free of M. avium subsp. paratuberculosis but located in the geographic vicinity of farms with infected livestock. PMID- 16269732 TI - Biocatalytic conversion of avermectin to 4"-oxo-avermectin: characterization of biocatalytically active bacterial strains and of cytochrome p450 monooxygenase enzymes and their genes. AB - 4"-Oxo-avermectin is a key intermediate in the manufacture of the agriculturally important insecticide emamectin benzoate from the natural product avermectin. Seventeen biocatalytically active Streptomyces strains with the ability to oxidize avermectin to 4"-oxo-avermectin in a regioselective manner have been discovered in a screen of 3,334 microorganisms. The enzymes responsible for this oxidation reaction in these biocatalytically active strains were found to be cytochrome P450 monooxygenases (CYPs) and were termed Ema1 to Ema17. The genes for Ema1 to Ema17 have been cloned, sequenced, and compared to reveal a new subfamily of CYPs. Ema1 to Ema16 have been overexpressed in Escherichia coli and purified as His-tagged recombinant proteins, and their basic enzyme kinetic parameters have been determined. PMID- 16269733 TI - Biocatalytic conversion of avermectin to 4"-oxo-avermectin: heterologous expression of the ema1 cytochrome P450 monooxygenase. AB - The cytochrome P450 monooxygenase Ema1 from Streptomyces tubercidicus R-922 and its homologs from closely related Streptomyces strains are able to catalyze the regioselective oxidation of avermectin into 4"-oxo-avermectin, a key intermediate in the manufacture of the agriculturally important insecticide emamectin benzoate (V. Jungmann, I. Molnar, P. E. Hammer, D. S. Hill, R. Zirkle, T. G. Buckel, D. Buckel, J. M. Ligon, and J. P. Pachlatko, Appl. Environ. Microbiol. 71:6968-6976, 2005). The gene for Ema1 has been expressed in Streptomyces lividans, Streptomyces avermitilis, and solvent-tolerant Pseudomonas putida strains using different promoters and vectors to provide biocatalytically competent cells. Replacing the extremely rare TTA codon with the more frequent CTG codon to encode Leu4 in Ema1 increased the biocatalytic activities of S. lividans strains producing this enzyme. Ferredoxins and ferredoxin reductases were also cloned from Streptomyces coelicolor and biocatalytic Streptomyces strains and tested in ema1 coexpression systems to optimize the electron transport towards Ema1. PMID- 16269734 TI - Comparison of microbial community compositions of two subglacial environments reveals a possible role for microbes in chemical weathering processes. AB - Viable microbes have been detected beneath several geographically distant glaciers underlain by different lithologies, but comparisons of their microbial communities have not previously been made. This study compared the microbial community compositions of samples from two glaciers overlying differing bedrock. Bulk meltwater chemistry indicates that sulfide oxidation and carbonate dissolution account for 90% of the solute flux from Bench Glacier, Alaska, whereas gypsum/anhydrite and carbonate dissolution accounts for the majority of the flux from John Evans Glacier, Ellesmere Island, Nunavut, Canada. The microbial communities were examined using two techniques: clone libraries and dot blot hybridization of 16S rRNA genes. Two hundred twenty-seven clones containing amplified 16S rRNA genes were prepared from subglacial samples, and the gene sequences were analyzed phylogenetically. Although some phylogenetic groups, including the Betaproteobacteria, were abundant in clone libraries from both glaciers, other well-represented groups were found at only one glacier. Group specific oligonucleotide probes were developed for two phylogenetic clusters that were of particular interest because of their abundance or inferred biochemical capabilities. These probes were used in quantitative dot blot hybridization assays with a range of samples from the two glaciers. In addition to shared phyla at both glaciers, each glacier also harbored a subglacial microbial population that correlated with the observed aqueous geochemistry. These results are consistent with the hypothesis that microbial activity is an important contributor to the solute flux from glaciers. PMID- 16269735 TI - Redox fluctuation structures microbial communities in a wet tropical soil. AB - Frequent high-amplitude redox fluctuation may be a strong selective force on the phylogenetic and physiological composition of soil bacterial communities and may promote metabolic plasticity or redox tolerance mechanisms. To determine effects of fluctuating oxygen regimens, we incubated tropical soils under four treatments: aerobic, anaerobic, 12-h oxic/anoxic fluctuation, and 4-day oxic/anoxic fluctuation. Changes in soil bacterial community structure and diversity were monitored with terminal restriction fragment length polymorphism (T-RFLP) fingerprints. These profiles were correlated with gross N cycling rates, and a Web-based phylogenetic assignment tool was used to infer putative community composition from multiple fragment patterns. T-RFLP ordinations indicated that bacterial communities from 4-day oxic/anoxic incubations were most similar to field communities, whereas those incubated under consistently aerobic or anaerobic regimens developed distinctly different molecular profiles. Terminal fragments found in field soils persisted either in 4-day fluctuation/aerobic conditions or in anaerobic/12-h treatments but rarely in both. Only 3 of 179 total fragments were ubiquitous in all soils. Soil bacterial communities inferred from in silico phylogenetic assignment appeared to be dominated by Actinobacteria (especially Micrococcus and Streptomycetes), "Bacilli," "Clostridia," and Burkholderia and lost significant diversity under consistently or frequently anoxic incubations. Community patterns correlated well with redox-sensitive processes such as nitrification, dissimilatory nitrate reduction to ammonium (DNRA), and denitrification but did not predict patterns of more general functions such as N mineralization and consumption. The results suggest that this soil's indigenous bacteria are highly adapted to fluctuating redox regimens and generally possess physiological tolerance mechanisms which allow them to withstand unfavorable redox periods. PMID- 16269736 TI - Bacterial community dynamics and polycyclic aromatic hydrocarbon degradation during bioremediation of heavily creosote-contaminated soil. AB - Bacterial community dynamics and biodegradation processes were examined in a highly creosote-contaminated soil undergoing a range of laboratory-based bioremediation treatments. The dynamics of the eubacterial community, the number of heterotrophs and polycyclic aromatic hydrocarbon (PAH) degraders, and the total petroleum hydrocarbon (TPH) and PAH concentrations were monitored during the bioremediation process. TPH and PAHs were significantly degraded in all treatments (72 to 79% and 83 to 87%, respectively), and the biodegradation values were higher when nutrients were not added, especially for benzo(a)anthracene and chrysene. The moisture content and aeration were determined to be the key factors associated with PAH bioremediation. Neither biosurfactant addition, bioaugmentation, nor ferric octate addition led to differences in PAH or TPH biodegradation compared to biodegradation with nutrient treatment. All treatments resulted in a high first-order degradation rate during the first 45 days, which was markedly reduced after 90 days. A sharp increase in the size of the heterotrophic and PAH-degrading microbial populations was observed, which coincided with the highest rates of TPH and PAH biodegradation. At the end of the incubation period, PAH degraders were more prevalent in samples to which nutrients had not been added. Denaturing gradient gel electrophoresis analysis and principal-component analysis confirmed that there was a remarkable shift in the composition of the bacterial community due to both the biodegradation process and the addition of nutrients. At early stages of biodegradation, the alpha Proteobacteria group (genera Sphingomonas and Azospirillum) was the dominant group in all treatments. At later stages, the gamma-Proteobacteria group (genus Xanthomonas), the alpha-Proteobacteria group (genus Sphingomonas), and the Cytophaga-Flexibacter-Bacteroides group (Bacteroidetes) were the dominant groups in the nonnutrient treatment, while the gamma-Proteobacteria group (genus Xathomonas), the beta-Proteobacteria group (genera Alcaligenes and Achromobacter), and the alpha-Proteobacteria group (genus Sphingomonas) were the dominant groups in the nutrient treatment. This study shows that specific bacterial phylotypes are associated both with different phases of PAH degradation and with nutrient addition in a preadapted PAH-contaminated soil. Our findings also suggest that there are complex interactions between bacterial species and medium conditions that influence the biodegradation capacity of the microbial communities involved in bioremediation processes. PMID- 16269737 TI - Culture-dependent and culture-independent diversity within the obligate marine actinomycete genus Salinispora. AB - Salinispora is the first obligate marine genus within the order Actinomycetales and a productive source of biologically active secondary metabolites. Despite a worldwide, tropical or subtropical distribution in marine sediments, only two Salinispora species have thus far been cultivated, suggesting limited species level diversity. To further explore Salinispora diversity and distributions, the phylogenetic diversity of more than 350 strains isolated from sediments collected around the Bahamas was examined, including strains cultured using new enrichment methods. A culture-independent method, using a Salinispora-specific seminested PCR technique, was used to detect Salinispora from environmental DNA and estimate diversity. Overall, the 16S rRNA gene sequence diversity of cultured strains agreed well with that detected in the environmental clone libraries. Despite extensive effort, no new species level diversity was detected, and 97% of the 105 strains examined by restriction fragment length polymorphism belonged to one phylotype (S. arenicola). New intraspecific diversity was detected in the libraries, including an abundant new phylotype that has yet to be cultured, and a new depth record of 1,100 m was established for the genus. PCR-introduced error, primarily from Taq polymerase, significantly increased clone library sequence diversity and, if not masked from the analyses, would have led to an overestimation of total diversity. An environmental DNA extraction method specific for vegetative cells provided evidence for active actinomycete growth in marine sediments while indicating that a majority of sediment samples contained predominantly Salinispora spores at concentrations that could not be detected in environmental clone libraries. Challenges involved with the direct sequence-based detection of spore-forming microorganisms in environmental samples are discussed. PMID- 16269738 TI - Inactivation of Bacillus endospores in envelopes by electron beam irradiation. AB - The anthrax incidents in the United States in the fall of 2001 led to the use of electron beam (EB) processing to sanitize the mail for the U.S. Postal Service. This method of sanitization has prompted the need to further investigate the effect of EB irradiation on the destruction of Bacillus endospores. In this study, endospores of an anthrax surrogate, B. atrophaeus, were destroyed to demonstrate the efficacy of EB treatment of such biohazard spores. EB exposures were performed to determine (i) the inactivation of varying B. atrophaeus spore concentrations, (ii) a D10 value (dose required to reduce a population by 1 log10) for the B. atrophaeus spores, (iii) the effects of spore survival at the bottom of a standardized paper envelope stack, and (iv) the maximum temperature received by spores. A maximum temperature of 49.2 degrees C was reached at a lethal dose of approximately 40 kGy, which is a significantly lower temperature than that needed to kill spores by thermal effects alone. A D10 value of 1.53 kGy was determined for the species. A surface EB dose between 25 and 32 kGy produced the appropriate killing dose of EB between 11 and 16 kGy required to inactivate 8 log10 spores, when spore samples were placed at the bottom of a 5.5-cm stack of envelopes. PMID- 16269739 TI - Evidence for a functional quorum-sensing type AI-1 system in the extremophilic bacterium Acidithiobacillus ferrooxidans. AB - Acidithiobacillus ferrooxidans is one of the main acidophilic chemolithotrophic bacteria involved in the bioleaching of metal sulfide ores. The bacterium-mineral interaction requires the development of biofilms, whose formation is regulated in many microorganisms by type AI-1 quorum sensing. Here, we report the existence and characterization of a functional type AI-1 quorum-sensing system in A. ferrooxidans. This microorganism produced mainly acyl-homoserine lactones (AHL) with medium and large acyl chains and different C-3 substitutions, including 3 hydroxy-C8-AHL, 3-hydroxy-C10-AHL, C12-AHL, 3-oxo-C12-AHL, 3-hydroxy-C12-AHL, C14 AHL, 3-oxo-C14-AHL, 3-hydroxy-C14-AHL, and 3-hydroxy-C16-AHL. A quorum-sensing genetic locus that includes two open reading frames, afeI and afeR, which have opposite orientations and code for proteins with high levels of similarity to members of the acyl synthase (I) and transcriptional regulator (R) protein families, respectively, was identified. Overexpression of AfeI in Escherichia coli and the associated synthesis of AHLs confirmed that AfeI is an AHL synthase. As determined by reverse transcription-PCR, the afeI and afeR genes were transcribed in A. ferrooxidans. The transcription levels of the afeI gene were higher in cells grown in sulfur and thiosulfate media than in iron-grown cells. Phosphate starvation induced an increase in the transcription levels of afeI which correlated with an increase in AHL levels. Two afe boxes which could correspond to the AfeR binding sites were identified upstream of the afeI gene. This is the first report of a functional type AI-1 quorum-sensing system in an acidophilic chemolithotrophic microorganism, and our results provide a very interesting opportunity to explore the control and regulation of biofilm formation during the bioleaching process. PMID- 16269740 TI - European origin of Bradyrhizobium populations infecting lupins and serradella in soils of Western Australia and South Africa. AB - We applied a multilocus phylogenetic approach to elucidate the origin of serradella and lupin Bradyrhizobium strains that persist in soils of Western Australia and South Africa. The selected strains belonged to different randomly amplified polymorphic DNA (RAPD)-PCR clusters that were distinct from RAPD clusters of applied inoculant strains. Phylogenetic analyses were performed with nodulation genes (nodA, nodZ, nolL, noeI), housekeeping genes (dnaK, recA, glnII, atpD), and 16S-23S rRNA intergenic transcribed spacer sequences. Housekeeping gene phylogenies revealed that all serradella and Lupinus cosentinii isolates from Western Australia and three of five South African narrow-leaf lupin strains were intermingled with the strains of Bradyrhizobium canariense, forming a well supported branch on each of the trees. All nodA gene sequences of the lupin and serradella bradyrhizobia formed a single branch, referred to as clade II, together with the sequences of other lupin and serradella strains. Similar patterns were detected in nodZ and nolL trees. In contrast, nodA sequences of the strains isolated from native Australian legumes formed either a new branch called clade IV or belonged to clade I or III, whereas their nonsymbiotic genes grouped outside the B. canariense branch. These data suggest that the lupin and serradella strains, including the strains from uncultivated L. cosentinii plants, are descendants of strains that most likely were brought from Europe accidentally with lupin and serradella seeds. The observed dominance of B. canariense strains may be related to this species' adaptation to acid soils common in Western Australia and South Africa and, presumably, to their intrinsic ability to compete for nodulation of lupins and serradella. PMID- 16269741 TI - Development of a cob-18S rRNA gene real-time PCR assay for quantifying Pfiesteria shumwayae in the natural environment. AB - Despite the fact that the heterotrophic dinoflagellate Pfiesteria shumwayae is an organism of high interest due to alleged toxicity, its abundance in natural environments is poorly understood. To address this inadequacy, a real-time quantitative PCR assay based on mitochondrial cytochrome b (cob) and 18S rRNA gene was developed and P. shumwayae abundance was investigated in several geographic locations. First, cob and its 5'-end region were isolated from a P. shumwayae culture, revealing three different copies, each consisting of an identical cob coding region and an unidentified region (X) of variable length and sequence. The unique sequences in cob and the X region were then used to develop a P. shumwayae-specific primer set. This primer set was used with reported P. shumwayae-specific 18S primers in parallel real-time PCRs to investigate P. shumwayae abundance from Maine to North Carolina along the U.S. east coast and along coasts in Chile, Hawaii, and China. Both genes generally gave similar results, indicating that this species was present, but at low abundance (mostly <10 cells x ml(-1)), in all the American coast locations investigated (with the exception of Long Island Sound, where which both genes gave negative results). Genetic variation was detected by use of both genes in most of the locations, and while cob consistently detected P. shumwayae or close genetic variants, some of the 18S PCR products were unrelated to P. shumwayae. We conclude that (i) the real-time PCR assay developed is useful for specific quantification of P. shumwayae, and (ii) P. shumwayae is distributed widely at the American coasts, but normally only as a minor component of plankton even in high-risk estuaries (Neuse River and the Chesapeake Bay). PMID- 16269742 TI - A method adapting microarray technology for signature-tagged mutagenesis of Desulfovibrio desulfuricans G20 and Shewanella oneidensis MR-1 in anaerobic sediment survival experiments. AB - Signature-tagged mutagenesis (STM) is a powerful technique that can be used to identify genes expressed by bacteria during exposure to conditions in their natural environments. To date, there have been no reports of studies in which this approach was used to study organisms of environmental, rather than pathogenic, significance. We used a mini-Tn10 transposon-bearing plasmid, pBSL180, that efficiently and randomly mutagenized Desulfovibrio desulfuricans G20 in addition to Shewanella oneidensis MR-1. Using these organisms as model sediment-dwelling anaerobic bacteria, we developed a new screening system, modified from former STM procedures, to identify genes that are critical for sediment survival. The screening system uses microarray technology to visualize tags from input and output pools, allowing us to identify those lost during sediment incubations. While the majority of data on survival genes identified will be presented in future papers, we report here on chemotaxis-related genes identified by our STM method in both bacteria in order to validate our method. This system may be applicable to the study of numerous environmental bacteria, allowing us to identify functions and roles of survival genes in various habitats. PMID- 16269743 TI - Survival of genetically modified and self-cloned strains of commercial baker's yeast in simulated natural environments: environmental risk assessment. AB - Although genetic engineering techniques for baker's yeast might improve the yeast's fermentation characteristics, the lack of scientific data on the survival of such strains in natural environments as well as the effects on human health prevent their commercial use. Disruption of acid trehalase gene (ATH1) improves freeze tolerance, which is a crucial characteristic in frozen-dough baking. In this study, ATH1 disruptants constructed by genetic modification (GM) and self cloning (SC) techniques were used as models to study such effects because these strains have higher freeze tolerance and are expected to be used commercially. Behavior of the strains in simulated natural environments, namely, in soil and water, was studied by measuring the change in the number of viable cells and in the concentration of DNA that contains ATH1 loci. Measurements were made using a real-time PCR method during 40 days of cultivation. Results showed that the number of viable cells of GM and SC strains decreased in a time-dependent manner and that the decrease rate was nearly equal to or higher than that for wild-type (WT) yeast. For all three strains (SC, GM, and WT) in the two simulated natural environments (water and soil), the DNA remained longer than did viable cells but the decrease patterns of either the DNA or the viable cells of SC and GM strains had tendencies similar to those of the WT strain. In conclusion, disruption of ATH1 by genetic engineering apparently does not promote the survival of viable cells and DNA in natural environments. PMID- 16269744 TI - New findings on evolution of metal homeostasis genes: evidence from comparative genome analysis of bacteria and archaea. AB - In order to examine the natural history of metal homeostasis genes in prokaryotes, open reading frames with homology to characterized P(IB)-type ATPases from the genomes of 188 bacteria and 22 archaea were investigated. Major findings were as follows. First, a high diversity in N-terminal metal binding motifs was observed. These motifs were distributed throughout bacterial and archaeal lineages, suggesting multiple loss and acquisition events. Second, the CopA locus separated into two distinct phylogenetic clusters, CopA1, which contained ATPases with documented Cu(I) influx activity, and CopA2, which contained both efflux and influx transporters and spanned the entire diversity of the bacterial domain, suggesting that CopA2 is the ancestral locus. Finally, phylogentic incongruences between 16S rRNA and P(IB)-type ATPase gene trees identified at least 14 instances of lateral gene transfer (LGT) that had occurred among diverse microbes. Results from bootstrapped supported nodes indicated that (i) a majority of the transfers occurred among proteobacteria, most likely due to the phylogenetic relatedness of these organisms, and (ii) gram-positive bacteria with low moles percent G+C were often involved in instances of LGT. These results, together with our earlier work on the occurrence of LGT in subsurface bacteria (J. M. Coombs and T. Barkay, Appl. Environ. Microbiol. 70:1698-1707, 2004), indicate that LGT has had a minor role in the evolution of P(IB)-type ATPases, unlike other genes that specify survival in metal-stressed environments. This study demonstrates how examination of a specific locus across microbial genomes can contribute to the understanding of phenotypes that are critical to the interactions of microbes with their environment. PMID- 16269745 TI - Kluyveromyces lactis LAC4 promoter variants that lack function in bacteria but retain full function in K. lactis. AB - The strong LAC4 promoter (P(LAC4)) from Kluyveromyces lactis has been extensively used to drive expression of heterologous proteins in this industrially important yeast. A drawback of this expression method is the serendipitous ability of P(LAC4) to promote gene expression in Escherichia coli. This can interfere with the process of assembling expression constructs in E. coli cells prior to their introduction into yeast cells, especially if the cloned gene encodes a protein that is detrimental to bacteria. In this study, we created a series of P(LAC4) variants by targeted mutagenesis of three DNA sequences (PBI, PBII, and PBIII) that resemble the E. coli Pribnow box element of bacterial promoters and that reside immediately upstream of two E. coli transcription initiation sites associated with P(LAC4). Mutation of PBI reduced the bacterial expression of a reporter protein (green fluorescent protein [GFP]) by approximately 87%, whereas mutation of PBII and PBIII had little effect on GFP expression. Deletion of all three sequences completely eliminated GFP expression. Additionally, each promoter variant expressed human serum albumin in K. lactis cells to levels comparable to wild-type P(LAC4). We created a novel integrative expression vector (pKLAC1) containing the P(LAC4) variant lacking PBI and used it to successfully clone and express the catalytic subunit of bovine enterokinase, a protease that has historically been problematic in E. coli cells. The pKLAC1 vector should aid in the cloning of other potentially toxic genes in E. coli prior to their expression in K. lactis. PMID- 16269746 TI - Stable coexistence of five bacterial strains as a cellulose-degrading community. AB - A cellulose-degrading defined mixed culture (designated SF356) consisting of five bacterial strains (Clostridium straminisolvens CSK1, Clostridium sp. strain FG4, Pseudoxanthomonas sp. strain M1-3, Brevibacillus sp. strain M1-5, and Bordetella sp. strain M1-6) exhibited both functional and structural stability; namely, no change in cellulose-degrading efficiency was observed, and all members stably coexisted through 20 subcultures. In order to investigate the mechanisms responsible for the observed stability, "knockout communities" in which one of the members was eliminated from SF356 were constructed. The dynamics of the community structure and the cellulose degradation profiles of these mixed cultures were determined in order to evaluate the roles played by each eliminated member in situ and its impact on the other members of the community. Integration of each result gave the following estimates of the bacterial relationships. Synergistic relationships between an anaerobic cellulolytic bacterium (C. straminisolvens CSK1) and two strains of aerobic bacteria (Pseudoxanthomonas sp. strain M1-3 and Brevibacillus sp. strain M1-5) were observed; the aerobes introduced anaerobic conditions, and C. straminisolvens CSK1 supplied metabolites (acetate and glucose). In addition, there were negative relationships, such as the inhibition of cellulose degradation by producing excess amounts of acetic acid by Clostridium sp. strain FG4, and growth suppression of Bordetella sp. strain M1-6 by Brevibacillus sp. strain M1-5. The balance of the various types of relationships (both positive and negative) is thus considered to be essential for the stable coexistence of the members of this mixed culture. PMID- 16269747 TI - Occurrence of Mycobacterium avium subsp. paratuberculosis in untreated water in Northern Ireland. AB - Mycobacterium avium subsp. paratuberculosis is the known cause of Johne's disease of both domestic and wild ruminants and has been implicated as a possible cause of Crohn's disease in humans. The organism is shed in the feces of infected animals and can survive for protracted periods in the environment and hence could be present in catchment areas receiving agricultural runoff. A limited survey was undertaken in Northern Ireland to test for M. avium subsp. paratuberculosis in untreated water entering nine water treatment works (WTWs) over a 1-year period. Three detection methods were employed, viz., immunomagnetic separation-PCR and culture on Herrold's egg yolk medium (HEYM) and BACTEC 12B medium, the latter both supplemented with mycobactins. Of the 192 untreated water samples tested, 15 (8%) tested M. avium subsp. paratuberculosis positive by one or more of the three detection methods. M. avium subsp. paratuberculosis was successfully isolated from eight untreated water samples, three by BACTEC culture and five by culture on HEYM. Although the highest incidence of M. avium subsp. paratuberculosis was found in spring, overall, there was no statistically significant difference between the seasons. No significant correlation was found between numbers of coliforms or fecal coliforms and the presence of M. avium subsp. paratuberculosis. In general, a higher incidence of M. avium subsp. paratuberculosis was found in untreated water entering those WTWs that had a high mean water pH value over the sampling period. This work indicates the need to determine the efficacy of water treatment processes to either kill or remove M. avium subsp. paratuberculosis from untreated water and the possible risks posed by contact with recreational water sources. PMID- 16269748 TI - Real-time nucleic acid sequence-based amplification assay for detection of hepatitis A virus. AB - A nucleic acid sequence-based amplification (NASBA) assay in combination with a molecular beacon was developed for the real-time detection and quantification of hepatitis A virus (HAV). A 202-bp, highly conserved 5' noncoding region of HAV was targeted. The sensitivity of the real-time NASBA assay was tested with 10 fold dilutions of viral RNA, and a detection limit of 1 PFU was obtained. The specificity of the assay was demonstrated by testing with other environmental pathogens and indicator microorganisms, with only HAV positively identified. When combined with immunomagnetic separation, the NASBA assay successfully detected as few as 10 PFU from seeded lake water samples. Due to its isothermal nature, its speed, and its similar sensitivity compared to the real-time RT-PCR assay, this newly reported real-time NASBA method will have broad applications for the rapid detection of HAV in contaminated food or water. PMID- 16269749 TI - Prevalence of pathogenic Yersinia enterocolitica strains in pigs in the United States. AB - Yersinia enterocolitica is considered an important food-borne pathogen impacting the pork production and processing industry in the United States. Since this bacterium is a commensal of swine, the primary goal of this study was to determine the prevalence of pathogenic Y. enterocolitica in pigs in the United States using feces as the sample source. A total of 2,793 fecal samples were tested for its presence in swine. Fecal samples were collected from late finisher pigs from 77 production sites in the 15 eastern and midwestern pork-producing states over a period of 27 weeks (6 September 2000 to 20 March 2001). The prevalence of ail-positive Y. enterocolitica was determined in samples using both a fluorogenic 5' nuclease PCR assay and a culture method. The mean prevalence was 13.10% (366 of 2,793 fecal samples tested) when both PCR- and culture-positive results were combined. Forty-one of 77 premises (53.25%) contained at least one fecal sample positive for the ail sequence. The PCR assay indicated a contamination rate of 12.35% (345/2,793) compared to 4.08% (114/2,793) by the culture method. Of the 345 PCR-positive samples, 252 were culture negative, while of the 114 culture-positive samples, 21 were PCR negative. Among 77 premises, the PCR assay revealed a significantly (P < 0.05) higher percentage (46.75%, n = 36 sites) of samples positive for the pathogen (ail sequence) than the culture method (22.08%, n = 17 sites). Thus, higher sensitivity, with respect to number of samples and sites identified as positive for the PCR method compared with the culture method for detecting pathogenic Y. enterocolitica, was demonstrated in this study. The results support the hypothesis that swine are a reservoir for Y. enterocolitica strains potentially pathogenic for humans. PMID- 16269750 TI - Engineering of Bacillus subtilis for enhanced total synthesis of folic acid. AB - We investigated whether the yield of the B vitamin folic acid could be elevated in Bacillus subtilis. Strategies for increasing the folic acid yield were investigated by employing computer-aided flux analysis and mutation. Controlling the activity of the enzyme pyruvate kinase by placing it under inducible control was one strategy devised to elevate yield while insuring that a rapid growth rate results. Other single mutation strategies included amplifying the expression of the genes in the folate operon and overexpressing the Escherichia coli aroH gene, which encodes 2-dehydro-3-deoxyphosphoheptonate aldolase. The latter could conceivably elevate the abundance of the folic acid precursor, para-aminobenzoic acid. Strains that combined two or more mutations were also constructed. Overall, a strain possessing inducible pyruvate kinase, overexpressed aroH, and increased transcription and translation of genes from the folic operon exhibited the best yield. The yield was eightfold higher than that displayed by the parent B. subtilis 168 strain. PMID- 16269751 TI - Novel DNA sequences from natural strains of the nitrogen-fixing symbiotic bacterium Sinorhizobium meliloti. AB - Variation in genome size and content is common among bacterial strains. Identifying these naturally occurring differences can accelerate our understanding of bacterial attributes, such as ecological specialization and genome evolution. In this study, we used representational difference analysis to identify potentially novel sequences not present in the sequenced laboratory strain Rm1021 of the nitrogen-fixing bacterium Sinorhizobium meliloti. Using strain Rm1021 as the driver and the type strain of S. meliloti ATCC 9930, which has a genome size approximately 370 kilobases bigger than that of strain Rm1021, as the tester, we identified several groups of sequences in the ATCC 9930 genome not present in strain Rm1021. Among the 85 novel DNA fragments examined, 55 showed no obvious homologs anywhere in the public databases. Of the remaining 30 sequences, 24 contained homologs to the Rm1021 genome as well as unique segments not found in Rm1021, 3 contained sequences homologous to those published for another S. meliloti strain but absent in Rm1021, 2 contained sequences homologous to other symbiotic nitrogen-fixing bacteria (Rhizobium etli and Bradyrhizobium japonicum), and 1 contained a sequence homologous to a gene in a non-nitrogen fixing species, Pseudomonas sp. NK87. Using PCR, we assayed the distribution of 12 of the above 85 novel sequences in a collection of 59 natural S. meliloti strains. The distribution varied widely among the 12 novel DNA fragments, from 1.7% to 72.9%. No apparent correlation was found between the distribution of these novel DNA sequences and their genotypes obtained using multilocus enzyme electrophoresis. Our results suggest potentially high rates of gene gain and loss in S. meliloti genomes. PMID- 16269752 TI - Metabolic engineering of Corynebacterium glutamicum for L-serine production. AB - Although L-serine proceeds in just three steps from the glycolytic intermediate 3 phosphoglycerate, and as much as 8% of the carbon assimilated from glucose is directed via L-serine formation, previous attempts to obtain a strain producing L serine from glucose have not been successful. We functionally identified the genes serC and serB from Corynebacterium glutamicum, coding for phosphoserine aminotransferase and phosphoserine phosphatase, respectively. The overexpression of these genes, together with the third biosynthetic serA gene, serA(delta197), encoding an L-serine-insensitive 3-phosphoglycerate dehydrogenase, yielded only traces of L-serine, as did the overexpression of these genes in a strain with the L-serine dehydratase gene sdaA deleted. However, reduced expression of the serine hydroxymethyltransferase gene glyA, in combination with the overexpression of serA(delta197), serC, and serB, resulted in a transient accumulation of up to 16 mM L-serine in the culture medium. When sdaA was also deleted, the resulting strain, C. glutamicum delta sdaA::pK18mobglyA'(pEC-T18mob2serA(delta197)CB), accumulated up to 86 mM L-serine with a maximal specific productivity of 1.2 mmol h(-1) g (dry weight)(-1). This illustrates a high rate of L-serine formation and also utilization in the C. glutamicum wild type. Therefore, metabolic engineering of L-serine production from glucose can be achieved only by addressing the apparent key position of this amino acid in the central metabolism. PMID- 16269753 TI - Transcriptional expression of the tceA gene in a Dehalococcoides-containing microbial enrichment. AB - Dynamic changes in the transcriptional expression of the tceA gene, which encodes a trichloroethene reductive dehalogenase, were characterized in a Dehalococcoides containing microbial enrichment culture. Expression was quantified by real-time PCR as the number of tceA transcripts per tceA gene. Expression of tceA increased 40-fold after chlorinated ethene-starved cells were exposed to trichloroethene (TCE), cis-dichloroethene (DCE), or 1,1-DCE but did not increase after exposure to tetrachloroethene or vinyl chloride. Surprisingly, tceA expression also increased 30-fold after cellular exposure to the nonmetabolic substrate trans DCE, indicating that expression of tceA is induced by both growth-supporting and non-growth-supporting chlorinated ethenes. Additional experiments revealed that the level of tceA expression was independent of the concentration of chlorinated ethenes (sum concentrations of TCE and DCEs of 2.2 to 333 microM), the concentration of the electron donor hydrogen (concentrations of 12 nM to 17 microM), and the presence of alternate bacterial electron acceptors (5 mM concentrations of fumarate, sulfate, sulfite, thiosulfate, nitrate, or nitrite) but was highly dependent on incubation temperature. PMID- 16269754 TI - Proteomic signature of Lactococcus lactis NCDO763 cultivated in milk. AB - We have compared the proteomic profiles of L. lactis subsp. cremoris NCDO763 growing in the synthetic medium M17Lac, skim milk microfiltrate (SMM), and skim milk. SMM was used as a simple model medium to reproduce the initial phase of growth of L. lactis in milk. To widen the analysis of the cytoplasmic proteome, we used two different gel systems (pH ranges of 4 to 7 and 4.5 to 5.5), and the proteins associated with the cell envelopes were also studied by two-dimensional electrophoresis. In the course of the study, we analyzed about 800 spots and identified 330 proteins by mass spectrometry. We observed that the levels of more than 50 and 30 proteins were significantly increased upon growth in SMM and milk, respectively. The large redeployment of protein synthesis was essentially associated with an activation of pathways involved in the metabolism of nitrogenous compounds: peptidolytic and peptide transport systems, amino acid biosynthesis and interconversion, and de novo biosynthesis of purines. We also showed that enzymes involved in reactions feeding the purine biosynthetic pathway in one-carbon units and amino acids have an increased level in SMM and milk. The analysis of the proteomic data suggested that the glutamine synthetase (GS) would play a pivotal role in the adaptation to SMM and milk. The analysis of glnA expression during growth in milk and the construction of a glnA-defective mutant confirmed that GS is an essential enzyme for the development of L. lactis in dairy media. This analysis thus provides a proteomic signature of L. lactis, a model lactic acid bacterium, growing in its technological environment. PMID- 16269755 TI - Algal species and light microenvironment in a low-pH, geothermal microbial mat community. AB - Unicellular algae are the predominant microbial mat-forming phototrophs in the extreme environments of acidic geothermal springs. The ecology of these algae is not well known because concepts of species composition are inferred from cultivated isolates and microscopic observations, methods known to provide incomplete and inaccurate assessments of species in situ. We used sequence analysis of 18S rRNA genes PCR amplified from mat samples from different seasons and different temperatures along a thermal gradient to identify algae in an often studied acidic (pH 2.7) geothermal creek in Yellowstone National Park. Fiber optic microprobes were used to show that light for algal photosynthesis is attenuated to < 1% over the 1-mm surface interval of the mat. Three algal sequences were detected, and each was present year-round. A Cyanidioschyzon merolae sequence was predominant at temperatures of > or = 49 degrees C. A Chlorella protothecoides var. acidicola sequence and a Paradoxia multisita-like sequence were predominant at temperatures of < or = 39 degrees C. PMID- 16269757 TI - Fate and role of ammonium ions during fermentation of citric acid by Aspergillus niger. AB - Stoichiometric modeling of the early stages of the citric acid fermentation process by Aspergillus niger revealed that ammonium ions combine with a carbon containing metabolite inside the cell, in a ratio 1:1, to form a nitrogen compound which is then excreted by the mycelium. High-performance liquid chromatography analysis identified glucosamine as the product of the relationship between glucose and ammonium during the early stages of the citric acid fermentation process. Slightly acidic internal pHs, extremely low ammonium ion concentrations inside the cell, and glucosamine synthesis come into direct contradiction with the earlier theory of the ammonium pool inside the cell, regarded as responsible for inhibition of the enzyme phosphofructokinase. At later fermentation stages, when the mycelium is involved in a process of fragmentation and regrowth, the addition of ammonium sulfate leads to a series of events: the formation and secretion of glucosamine in elevated amounts, the short inhibition of citrate synthesis, growth enhancement, the utilization of glucosamine, and finally, the enhancement of citric acid production rates. Obviously, the enzymatic processes underlining the phenomena need to be reexamined. As a by-product of the citric acid fermentation, glucosamine is reported for the first time here. Suitable process manipulations of the system described in this work could lead to successful glucosamine recovery at the point of its highest yield before degradation by the fungus occurs. PMID- 16269756 TI - Effect of oxidation rate and Fe(II) state on microbial nitrate-dependent Fe(III) mineral formation. AB - A nitrate-dependent Fe(II)-oxidizing bacterium was isolated and used to evaluate whether Fe(II) chemical form or oxidation rate had an effect on the mineralogy of biogenic Fe(III) (hydr)oxides resulting from nitrate-dependent Fe(II) oxidation. The isolate (designated FW33AN) had 99% 16S rRNA sequence similarity to Klebsiella oxytoca. FW33AN produced Fe(III) (hydr)oxides by oxidation of soluble Fe(II) [Fe(II)sol] or FeS under nitrate-reducing conditions. Based on X-ray diffraction (XRD) analysis, Fe(III) (hydr)oxide produced by oxidation of FeS was shown to be amorphous, while oxidation of Fe(II)sol yielded goethite. The rate of Fe(II) oxidation was then manipulated by incubating various cell concentrations of FW33AN with Fe(II)sol and nitrate. Characterization of products revealed that as Fe(II) oxidation rates slowed, a stronger goethite signal was observed by XRD and a larger proportion of Fe(III) was in the crystalline fraction. Since the mineralogy of Fe(III) (hydr)oxides may control the extent of subsequent Fe(III) reduction, the variables we identify here may have an effect on the biogeochemical cycling of Fe in anoxic ecosystems. PMID- 16269758 TI - Generation and molecular characterization of new temperature-sensitive plasmids intended for genetic engineering of Pasteurellaceae. AB - Temperature-sensitive (TS) plasmids were generated through chemical mutagenesis of a derivative of the streptomycin resistance parent plasmid pD70, isolated from Mannheimia hemolytica serotype 1. Three TS plasmids which failed to replicate at or above 42 degrees C in M. hemolytica but which were fully functional below 31 degrees C were selected for further analysis. Two of the TS plasmids were shown by sequencing to possess unique single-base-pair mutations. The third TS plasmid contained a unique base pair substitution and a second mutation that had been previously identified. These mutations were clustered within a 200-bp region of the presumed plasmid origin of replication. Site-directed single-nucleotide substitutions were introduced into the wild-type pD70 origin of replication to confirm that mutations identified by sequencing had conferred thermoregulated replication. Deletion analysis on the wild-type pD70 plasmid replicon revealed that approximately 720 bp are necessary for plasmid maintenance. Replication of the TS plasmids was thermoregulated in Pasteurella multocida and Haemophilus somnus as well. To consistently transform H. somnus with TS plasmid, in vitro DNA methylation with commercially available HhaI methyltransferase was necessary to protect against the organism's restriction enzyme HsoI (recognition sequence 5' GCGC-3') characterized herein. PMID- 16269759 TI - Construction of in-frame aroA deletion mutants of Mannheimia haemolytica, Pasteurella multocida, and Haemophilus somnus by using a new temperature sensitive plasmid. AB - A temperature-sensitive (TS) plasmid was generated from the endogenous streptomycin resistance plasmid of Mannheimia hemolytica and used to engineer in frame aroA deletion mutants of Mannheimia hemolytica, Pasteurella multocida, and Haemophilus somnus. TS replacement plasmids carrying in-frame aroA deletions were constructed for each target species and introduced into host cells by electroporation. After recovery in broth, cells were spread onto plates containing antibiotic and incubated at 30 degrees C, the permissive temperature for autonomous plasmid replication. Transfer of transformants to selective plates cultured at a nonpermissive temperature for plasmid replication selected for single-crossover mutants consisting of replacement plasmids that had integrated into host chromosomes by homologous recombination. Transfer of the single crossover mutants back to a permissive temperature without antibiotic selection drove plasmid resolution, and, depending on where plasmid excision occurred, either deletion mutants or wild-type cells were generated. The system used here represents a broadly applicable means for generating unmarked mutants of Pasteurellaceae species. PMID- 16269761 TI - 16S rRNA gene-based identification of midgut bacteria from field-caught Anopheles gambiae sensu lato and A. funestus mosquitoes reveals new species related to known insect symbionts. AB - Field-collected mosquitoes of the two main malaria vectors in Africa, Anopheles gambiae sensu lato and Anopheles funestus, were screened for their midgut bacterial contents. The midgut from each blood-fed mosquito was screened with two different detection pathways, one culture independent and one culture dependent. Bacterial species determination was achieved by sequence analysis of 16S rRNA genes. Altogether, 16 species from 14 genera were identified, 8 by each method. Interestingly, several of the bacteria identified are related to bacteria known to be symbionts in other insects. One isolate, Nocardia corynebacterioides, is a relative of the symbiont found in the vector for Chagas' disease that has been proven useful as a paratransgenic bacterium. Another isolate is a novel species within the gamma-proteobacteria that could not be phylogenetically placed within any of the known orders in the class but is close to a group of insect symbionts. Bacteria representing three intracellular genera were identified, among them the first identifications of Anaplasma species from mosquitoes and a new mosquito Spiroplasma association. The isolates will be further investigated for their suitability for a paratransgenic Anopheles mosquito. PMID- 16269760 TI - Prevalence of Borrelia burgdorferi sensu lato genospecies in Ixodes ricinus ticks in Europe: a metaanalysis. AB - In Europe, Borrelia burgdorferi genospecies causing Lyme borreliosis are mainly transmitted by the tick Ixodes ricinus. Since its discovery, B. burgdorferi has been the subject of many epidemiological studies to determine its prevalence and the distribution of the different genospecies in ticks. In the current study we systematically reviewed the literature on epidemiological studies of I. ricinus ticks infected with B. burgdorferi sensu lato. A total of 1,186 abstracts in English published from 1984 to 2003 were identified by a PubMed keyword search and from the compiled article references. A multistep filter process was used to select relevant articles; 110 articles from 24 countries contained data on the rates of infection of I. ricinus with Borrelia in Europe (112,579 ticks), and 44 articles from 21 countries included species-specific analyses (3,273 positive ticks). These data were used to evaluate the overall rate of infection of I. ricinus with Borrelia genospecies, regional distributions within Europe, and changes over time, as well as the influence of different detection methods on the infection rate. While the infection rate was significantly higher in adults (18.6%) than in nymphs (10.1%), no effect of detection method, tick gender, or collection period (1986 to 1993 versus 1994 to 2002) was found. The highest rates of infection of I. ricinus were found in countries in central Europe. B. afzelii and B. garinii are the most common Borrelia species, but the distribution of genospecies seems to vary in different regions in Europe. The most frequent coinfection by Borrelia species was found for B. garinii and B. valaisiana. PMID- 16269762 TI - Feedback inhibition of chorismate mutase/prephenate dehydrogenase (TyrA) of Escherichia coli: generation and characterization of tyrosine-insensitive mutants. AB - In order to get insights into the feedback regulation by tyrosine of the Escherichia coli chorismate mutase/prephenate dehydrogenase (CM/PDH), which is encoded by the tyrA gene, feedback-inhibition-resistant (fbr) mutants were generated by error-prone PCR. The tyrA(fbr) mutants were selected by virtue of their resistance toward m-fluoro-D,L-tyrosine, and seven representatives were characterized on the biochemical as well as on the molecular level. The PDH activities of the purified His6-tagged TyrA proteins exhibited up to 35% of the enzyme activity of TyrA(WT), but tyrosine did not inhibit the mutant PDH activities. On the other hand, CM activities of the TyrA(fbr) mutants were similar to those of the TyrA(WT) protein. Analyses of the DNA sequences of the tyrA genes revealed that tyrA(fbr) contained amino acid substitutions either at Tyr263 or at residues 354 to 357, indicating that these two sites are involved in the feedback inhibition by tyrosine. PMID- 16269763 TI - Alteration of leucine aminopeptidase from Streptomyces septatus TH-2 to phenylalanine aminopeptidase by site-directed mutagenesis. AB - To tailor leucine aminopeptidase from Streptomyces septatus TH-2 (SSAP) to become a convenient biocatalyst, we are interested in Phe221 of SSAP, which is thought to interact with the side chain of the N-terminal residue of the substrate. By using saturation mutagenesis, the feasibility of altering the performance of SSAP was evaluated. The hydrolytic activities of 19 mutants were investigated using aminoacyl p-nitroanilide (pNA) derivatives as substrates. Replacement of Phe221 resulted in changes in the activities of all the mutants. Three of these mutants, F221G, F221A, and F221S, specifically hydrolyzed L-Phe-pNA, and F221I SSAP exhibited hydrolytic activity with L-Leu-pNA exceeding that of the wild type. Although the hydrolytic activities with peptide substrates decreased, the hydrolytic activities with amide and methyl ester substrates were proportional to the changes in the hydrolytic activities with pNA derivatives. Furthermore, based on a comparative kinetic study, the mechanism underlying the alteration in the preference of SSAP from leucine to phenylalanine is discussed. PMID- 16269764 TI - Recognition of individual genes in diverse microorganisms by cycling primed in situ amplification. AB - Cycling primed in situ amplification-fluorescent in situ hybridization (CPRINS FISH) was developed to recognize individual genes in a single bacterial cell. In CPRINS, the amplicon was long single-stranded DNA and thus retained within the permeabilized microbial cells. FISH with a multiply labeled fluorescent probe set enabled significant reduction in nonspecific background while maintaining high fluorescence signals of target bacteria. The ampicillin resistance gene in Escherichia coli, chloramphenicol acetyltransferase gene in different gram negative strains, and RNA polymerase sigma factor (rpoD) gene in Aeromonas spp. could be detected under identical permeabilization conditions. After concentration of environmental freshwater samples onto polycarbonate filters and subsequent coating of filters in gelatin, no decrease in bacterial cell numbers was observed with extensive permeabilization. The detection rates of bacterioplankton in river and pond water samples by CPRINS-FISH with a universal 16S rRNA gene primer and probe set ranged from 65 to 76% of total cell counts (mean, 71%). The concentrations of cells detected by CPRINS-FISH targeting of the rpoD genes of Aeromonas sobria and A. hydrophila in the water samples varied between 2.1 x 10(3) and 9.0 x 10(3) cells ml(-1) and between undetectable and 5.1 x 10(2) cells ml(-1), respectively. These results demonstrate that CPRINS-FISH provides a high sensitivity for microscopic detection of bacteria carrying a specific gene in natural aquatic samples. PMID- 16269765 TI - Methylotrophic metabolism is advantageous for Methylobacterium extorquens during colonization of Medicago truncatula under competitive conditions. AB - Facultative methylotrophic bacteria of the genus Methylobacterium are commonly found in association with plants. Inoculation experiments were performed to study the importance of methylotrophic metabolism for colonization of the model legume Medicago truncatula. Competition experiments with Methylobacterium extorquens wild-type strain AM1 and methylotrophy mutants revealed that the ability to use methanol as a carbon and energy source provides a selective advantage during colonization of M. truncatula. Differences in the fitness of mutants defective in different stages of methylotrophic metabolism were found; whereas approximately 25% of the mutant incapable of oxidizing methanol to formaldehyde (deficient in methanol dehydrogenase) was recovered, 10% or less of the mutants incapable of oxidizing formaldehyde to CO2 (defective in biosynthesis of the cofactor tetrahydromethanopterin) was recovered. Interestingly, impaired fitness of the mutant strains compared with the wild type was found on leaves and roots. Single inoculation experiments showed, however, that mutants with defects in methylotrophy were capable of plant colonization at the wild-type level, indicating that methanol is not the only carbon source that is accessible to Methylobacterium while it is associated with plants. Fluorescence microscopy with a green fluorescent protein-labeled derivative of M. extorquens AM1 revealed that the majority of the bacterial cells on leaves were on the surface and that the cells were most abundant on the lower, abaxial side. However, bacterial cells were also found in the intercellular spaces inside the leaves, especially in the epidermal cell layer and immediately underneath this layer. PMID- 16269766 TI - In silico reconstruction of the metabolic pathways of Lactobacillus plantarum: comparing predictions of nutrient requirements with those from growth experiments. AB - On the basis of the annotated genome we reconstructed the metabolic pathways of the lactic acid bacterium Lactobacillus plantarum WCFS1. After automatic reconstruction by the Pathologic tool of Pathway Tools (http://bioinformatics.ai.sri.com/ptools/), the resulting pathway-genome database, LacplantCyc, was manually curated extensively. The current database contains refinements to existing routes and new gram-positive bacterium-specific reactions that were not present in the MetaCyc database. These reactions include, for example, reactions related to cell wall biosynthesis, molybdopterin biosynthesis, and transport. At present, LacplantCyc includes 129 pathways and 704 predicted reactions involving some 670 chemical species and 710 enzymes. We tested vitamin and amino acid requirements of L. plantarum experimentally and compared the results with the pathways present in LacplantCyc. In the majority of cases (32 of 37 cases) the experimental results agreed with the final reconstruction. LacplantCyc is the most extensively curated pathway-genome database for gram-positive bacteria and is open to the microbiology community via the World Wide Web (www.lacplantcyc.nl). It can be used as a reference pathway genome database for gram-positive microbes in general and lactic acid bacteria in particular. PMID- 16269768 TI - Ascending migration of endophytic rhizobia, from roots to leaves, inside rice plants and assessment of benefits to rice growth physiology. AB - Rhizobia, the root-nodule endosymbionts of leguminous plants, also form natural endophytic associations with roots of important cereal plants. Despite its widespread occurrence, much remains unknown about colonization of cereals by rhizobia. We examined the infection, dissemination, and colonization of healthy rice plant tissues by four species of gfp-tagged rhizobia and their influence on the growth physiology of rice. The results indicated a dynamic infection process beginning with surface colonization of the rhizoplane (especially at lateral root emergence), followed by endophytic colonization within roots, and then ascending endophytic migration into the stem base, leaf sheath, and leaves where they developed high populations. In situ CMEIAS image analysis indicated local endophytic population densities reaching as high as 9 x 10(10) rhizobia per cm3 of infected host tissues, whereas plating experiments indicated rapid, transient or persistent growth depending on the rhizobial strain and rice tissue examined. Rice plants inoculated with certain test strains of gfp-tagged rhizobia produced significantly higher root and shoot biomass; increased their photosynthetic rate, stomatal conductance, transpiration velocity, water utilization efficiency, and flag leaf area (considered to possess the highest photosynthetic activity); and accumulated higher levels of indoleacetic acid and gibberellin growth-regulating phytohormones. Considered collectively, the results indicate that this endophytic plant-bacterium association is far more inclusive, invasive, and dynamic than previously thought, including dissemination in both below-ground and above-ground tissues and enhancement of growth physiology by several rhizobial species, therefore heightening its interest and potential value as a biofertilizer strategy for sustainable agriculture to produce the world's most important cereal crops. PMID- 16269767 TI - Ecology, inhibitory activity, and morphogenesis of a marine antagonistic bacterium belonging to the Roseobacter clade. AB - Roseobacter strain 27-4 has been isolated from a turbot larval rearing unit and is capable of reducing mortality in turbot egg yolk sac larvae. Here, we demonstrate that the supernatant of Roseobacter 27-4 is lethal to the larval pathogens Vibrio anguillarum and Vibrio splendidus in a buffer system and inhibited their growth in marine broth. Liquid chromatography (LC) with both UV spectral detection and high-resolution mass spectrometry (HR-MS) identified the known antibacterial compound thiotropocin or its closely related precursor tropodithietic acid in the bioactive fractions. Antibacterial activity correlated with the appearance of a brownish pigment and was only formed in marine broth under static growth conditions. A thick biofilm of multicellular star-shaped aggregated cells formed at the air-liquid interface under static growth conditions. Here, the bioactive compound was the base peak in the LC-UV chromatograms of the extracts where it constituted 15% of the total peak area. Aerated conditions results in 10-fold-higher cell yield, however, cultures were nonpigmented, did not produce antibacterial activity, and grew as single cells. Production of antibacterial compounds may be quorum regulated, and we identified the acylated homoserine lactone (3-hydroxy-decanoyl homoserine lactone) from cultures of Roseobacter 27-4 using LC-HR-MS. The signal molecule was primarily detected in stagnant cultures. Roseobacter 27-4 grew between 10 and 30 degrees C but died rapidly at 37 degrees C. Also, the antibacterial compounds was sensitive to heat and was inactivated at 37 degrees C in less than 2 days and at 25 degrees C in 8 days. Using Roseobacter 27-4 as a probiotic culture will require that is be established in stagnant or adhered conditions and, due to the temperature sensitivity of the active compound, constant production must be ensured. PMID- 16269769 TI - Copper-adapted Suillus luteus, a symbiotic solution for pines colonizing Cu mine spoils. AB - Natural populations thriving in heavy-metal-contaminated ecosystems are often subjected to selective pressures for increased resistance to toxic metals. In the present study we describe a population of the ectomycorrhizal fungus Suillus luteus that colonized a toxic Cu mine spoil in Norway. We hypothesized that this population had developed adaptive Cu tolerance and was able to protect pine trees against Cu toxicity. We also tested for the existence of cotolerance to Cu and Zn in S. luteus. Isolates from Cu-polluted, Zn-polluted, and nonpolluted sites were grown in vitro on Cu- or Zn-supplemented medium. The Cu mine isolates exhibited high Cu tolerance, whereas the Zn-tolerant isolates were shown to be Cu sensitive, and vice versa. This indicates the evolution of metal-specific tolerance mechanisms is strongly triggered by the pollution in the local environment. Cotolerance does not occur in the S. luteus isolates studied. In a dose-response experiment, the Cu sensitivity of nonmycorrhizal Pinus sylvestris seedlings was compared to the sensitivity of mycorrhizal seedlings colonized either by a Cu-sensitive or Cu-tolerant S. luteus isolate. In nonmycorrhizal plants and plants colonized by the Cu-sensitive isolate, root growth and nutrient uptake were strongly inhibited under Cu stress conditions. In contrast, plants colonized by the Cu-tolerant isolate were hardly affected. The Cu-adapted S. luteus isolate provided excellent insurance against Cu toxicity in pine seedlings exposed to elevated Cu levels. Such a metal-adapted Suillus-Pinus combination might be suitable for large-scale land reclamation at phytotoxic metalliferous and industrial sites. PMID- 16269770 TI - Detection of carp interstitial nephritis and gill necrosis virus in fish droppings. AB - Carp interstitial nephritis and gill necrosis virus (CNGV) is an unclassified large DNA virus that morphologically resembles members of the Herpesviridae but contains a large (ca. approximately 280-kbp) linear double-stranded DNA. This virus has also been named koi herpesvirus, koi herpes-like virus, and cyprinid herpesvirus 3. CNGV is the cause of a lethal disease that afflicts common carp and koi. By using immunohistochemistry, molecular analysis, and electron microscopy we previously demonstrated that this virus is present mainly in the intestine and kidney of infected fish. Based on these observations, we postulated that viruses and/or viral components may appear in droppings of infected carp. Here we report that (i) by using PCR we demonstrated that fish droppings contain viral DNA, (ii) fish droppings contain viral antigens which are useful for CNGV diagnosis, and (iii) fish droppings contain active virus which can infect cultured common carp brain cells and induce the disease in naive fish following inoculation. Thus, our findings show that CNGV can be identified by using droppings without taking biopsies or killing fish and that infectious CNGV is present in the stools of sick fish. The possibility that fish droppings preserve viable CNGV during the nonpermissive seasons is discussed. PMID- 16269771 TI - Paenibacillus polymyxa invades plant roots and forms biofilms. AB - Paenibacillus polymyxa is a plant growth-promoting rhizobacterium with a broad host range, but so far the use of this organism as a biocontrol agent has not been very efficient. In previous work we showed that this bacterium protects Arabidopsis thaliana against pathogens and abiotic stress (S. Timmusk and E. G. H. Wagner, Mol. Plant-Microbe Interact. 12:951-959, 1999; S. Timmusk, P. van West, N. A. R. Gow, and E. G. H. Wagner, p. 1-28, in Mechanism of action of the plant growth promoting bacterium Paenibacillus polymyxa, 2003). Here, we studied colonization of plant roots by a natural isolate of P. polymyxa which had been tagged with a plasmid-borne gfp gene. Fluorescence microscopy and electron scanning microscopy indicated that the bacteria colonized predominantly the root tip, where they formed biofilms. Accumulation of bacteria was observed in the intercellular spaces outside the vascular cylinder. Systemic spreading did not occur, as indicated by the absence of bacteria in aerial tissues. Studies were performed in both a gnotobiotic system and a soil system. The fact that similar observations were made in both systems suggests that colonization by this bacterium can be studied in a more defined system. Problems associated with green fluorescent protein tagging of natural isolates and deleterious effects of the plant growth-promoting bacteria are discussed. PMID- 16269772 TI - Three stages of a biofilm community developing at the liquid-liquid interface between polychlorinated biphenyls and water. AB - Soil contaminated with polychlorinated biphenyls (PCB) was used as an inoculum to grow a complex biofilm community on PCB oil (Aroclor 1242) on a substratum (Permanox). The biofilm was monitored for 31 days by confocal laser scanning microscopy, community fingerprinting using single-strand conformational polymorphism (SSCP), amplicons of the 16S rRNA genes, and chemical analyses of the PCB congeners. SSCP analysis of the young biofilm revealed a rather diverse microbial community with species of the genera Herbaspirillum and Bradyrhizobium as dominant members. The biofilm developing on the PCB droplets displayed pronounced stages of PCB degradation and biofilm development not described before from pure-culture experiments. The first step was the colonization of the substratum while the PCB oil was hardly populated. When a certain density of bacteria was reached on the Permanox, the PCB was colonized, but soon the degradation of the congeners was markedly reduced and many cells were damaged, as seen by LIVE/DEAD staining. Finally, the biofilm formed aggregates and invaded the PCB oil, showing lower numbers of damaged cells than before and a dramatic increase in PCB degradation. This sequence of biofilm formation is understood as a maturation process prior to PCB oil colonization. This is followed by a thin biofilm on the PCB droplet, an aggregation process forming pockets in the PCB, and finally an invasion of the biofilm into the PCB oil. Only the mature biofilm showed degradation of pentachlorinated PCB congeners, which may be reductively dechlorinated and the resulting trichlorobiphenyls then aerobically metabolized. PMID- 16269773 TI - Enzymatic and genetic characterization of carbon and energy metabolisms by deep sea hydrothermal chemolithoautotrophic isolates of Epsilonproteobacteria. AB - The carbon and energy metabolisms of a variety of cultured chemolithoautotrophic Epsilonproteobacteria from deep-sea hydrothermal environments were characterized by both enzymatic and genetic analyses. All the Epsilonproteobacteria tested had all three key reductive tricarboxylic acid (rTCA) cycle enzymatic activities--ATP dependent citrate lyase, pyruvate:ferredoxin oxidoreductase, and 2 oxoglutarate:ferredoxin oxidoreductase--while they had no ribulose 1,5 bisphosphate carboxylase (RubisCO) activity, the key enzyme in the Calvin-Benson cycle. These results paralleled the successful amplification of the key rTCA cycle genes aclB, porAB, and oorAB and the lack of success at amplifying the form I and II RubisCO genes, cbbL and cbbM. The combination of enzymatic and genetic analyses demonstrates that the Epsilonproteobacteria tested use the rTCA cycle for carbon assimilation. The energy metabolisms of deep-sea Epsilonproteobacteria were also well specified by the enzymatic and genetic characterization: hydrogen oxidizing strains had evident soluble acceptor:methyl viologen hydrogenase activity and hydrogen uptake hydrogenase genes (hyn operon), while sulfur oxidizing strains lacked both the enzyme activity and the genes. Although the energy metabolism of reduced sulfur compounds was not genetically analyzed and was not fully clarified, sulfur-oxidizing Epsilonproteobacteria showed enzyme activity of a potential sulfite:acceptor oxidoreductase for a direct oxidation pathway to sulfate but no activity of AMP-dependent adenosine 5'-phosphate sulfate reductase for a indirect oxidation pathway. No activity of thiosulfate oxidizing enzymes was detected. The enzymatic and genetic characteristics described here were consistent with cellular carbon and energy metabolisms and suggest that molecular tools may have great potential for in situ elucidation of the ecophysiological roles of deep-sea Epsilonproteobacteria. PMID- 16269774 TI - Catalyzed reported deposition-fluorescence in situ hybridization protocol to evaluate phagotrophy in mixotrophic protists. AB - We describe a catalyzed reported deposition-fluorescence in situ hybridization (CARD-FISH) protocol particularly suited to assess the phagotrophy of mixotrophic protists on prokaryotes, since it maintains cell and plastid integrity, avoids cell loss and egestion of prey, and allows visualization of labeled prey against plastid autofluorescence. This protocol, which includes steps such as Lugol's formaldehyde-thiosulfate fixation, agarose cell attachment, cell wall permeabilization with lysozyme plus achromopeptidase, and signal amplification with Alexa-Fluor 488, allowed us to detect almost 100% of planktonic prokaryotes (Bacteria and Archaea) and, for the first time, to show archaeal cells ingested by mixotrophic protists. PMID- 16269775 TI - Crucial role of extracellular polysaccharides in desiccation and freezing tolerance in the terrestrial cyanobacterium Nostoc commune. AB - The cyanobacterium Nostoc commune is adapted to the terrestrial environment and has a cosmopolitan distribution. In this study, the role of extracellular polysaccharides (EPS) in the desiccation tolerance of photosynthesis in N. commune was examined. Although photosynthetic O2 evolution was not detected in desiccated colonies, the ability of the cells to evolve O2 rapidly recovered after rehydration. The air-dried colonies contained approximately 10% (wt/wt) water, and field-isolated, natural colonies with EPS were highly water absorbent and were rapidly hydrated by atmospheric moisture. The cells embedded in EPS in Nostoc colonies were highly desiccation tolerant, and O2 evolution was not damaged by air drying. Although N. commune was determined to be a mesophilic cyanobacterium, the cells with EPS were heat tolerant in a desiccated state. EPS could be removed from cells by homogenizing colonies with a blender and filtering with coarse filter paper. This treatment to remove EPS did not damage Nostoc cells or their ability to evolve O2, but O2 evolution was significantly damaged by desiccation treatment of the EPS-depleted cells. Similar to the EPS-depleted cells, the laboratory culture strain KU002 had only small amount of EPS and was highly sensitive to desiccation. In the EPS-depleted cells, O2 evolution was also sensitive to freeze-thaw treatment. These results strongly suggest that EPS of N. commune is crucial for the stress tolerance of photosynthesis during desiccation and during freezing and thawing. PMID- 16269776 TI - Antagonists of Hsp16.3, a low-molecular-weight mycobacterial chaperone and virulence factor, derived from phage-displayed peptide libraries. AB - The persistence of Mycobacterium tuberculosis is a major cause of concern in tuberculosis (TB) therapy. In the persistent mode the pathogen can resist drug therapy, allowing the possibility of reactivation of the disease. Several protein factors have been identified that contribute to persistence, one of them being the 16-kDa low-molecular-weight mycobacterial heat shock protein Hsp16.3, a homologue of the mammalian eye lens protein alpha-crystallin. It is believed that Hsp16.3 plays a key role in the persistence phase by protecting essential proteins from being irreversibly denatured. Because of the close association of Hsp16.3 with persistence, an attempt has been made to develop inhibitors against it. Random peptide libraries displayed on bacteriophage M13 were screened for Hsp16.3 binding. Two phage clones were identified that bind to the Hsp16.3 protein. The corresponding synthetic peptides, an 11-mer and a 16-mer, were able to bind Hsp16.3 and inhibit its chaperone activity in vitro in a dose-dependent manner. Little or no effect of these peptides was observed on alphaB-crystallin, a homologous protein that is a key component of human eye lens, indicating that there is an element of specificity in the observed inhibition. Two histidine residues appear to be common to the selected peptides. Nuclear magnetic resonance studies performed with the 11-mer peptide indicate that in this case these two histidines may be the crucial binding determinants. The peptide inhibitors of Hsp16.3 thus obtained could serve as the basis for developing potent drugs against persistent TB. PMID- 16269777 TI - Capsular polysaccharide surrounds smooth and rugose types of Salmonella enterica serovar Typhimurium DT104. AB - The biofilms and rugose colony morphology of Salmonella enterica serovar Typhimurium strains are usually associated with at least two different exopolymeric substances (EPS), curli and cellulose. In this study, another EPS, a capsular polysaccharide (CP) synthesized constitutively in S. enterica serovar Typhimurium strain DT104 at 25 and 37 degrees C, has been recognized as a biofilm matrix component as well. Fluorophore-assisted carbohydrate electrophoresis (FACE) analysis indicated that the CP is comprised principally of glucose and mannose, with galactose as a minor constituent. The composition differs from that of known colanic acid-containing CP that is isolated from cells of Escherichia coli and other enteric bacteria grown at 37 degrees C. The reactivity of carbohydrate-specific lectins conjugated to fluorescein isothiocyanate or gold particles with cellular carbohydrates demonstrated the cell surface localization of CP. Further, lectin binding also correlated with the FACE analysis of CP. Immunoelectron microscopy, using specific antibodies against CP, confirmed that CP surrounds the cells. Confocal microscopy of antibody-labeled cells showed greater biofilm formation at 25 degrees C than at 37 degrees C. Since the CP was shown to be produced at both 37 degrees C and 25 degrees C, it does not appear to be significantly involved in attachment during the early formation of the biofilm matrix. Although the attachment of S. enterica serovar Typhimurium DT104 does not appear to be mediated by its CP, the capsule does contribute to the biofilm matrix and may have a role in other features of this organism, such as virulence, as has been shown previously for the capsules of other gram-negative and gram positive bacteria. PMID- 16269778 TI - Community composition of a hypersaline endoevaporitic microbial mat. AB - A hypersaline, endoevaporitic microbial community in Eilat, Israel, was studied by microscopy and by PCR amplification of genes for 16S rRNA from different layers. In terms of biomass, the oxygenic layers of the community were dominated by Cyanobacteria of the Halothece, Spirulina, and Phormidium types, but cell counts (based on 4',6'-diamidino-2-phenylindole staining) and molecular surveys (clone libraries of PCR-amplified genes for 16S rRNA) showed that oxygenic phototrophs were outnumbered by the other constituents of the community, including chemotrophs and anoxygenic phototrophs. Bacterial clone libraries were dominated by phylotypes affiliated with the Bacteroidetes group and both photo- and chemotrophic groups of alpha-proteobacteria. Green filaments related to the Chloroflexi were less abundant than reported from hypersaline microbial mats growing at lower salinities and were only detected in the deepest part of the anoxygenic phototrophic zone. Also detected were nonphototrophic gamma- and delta proteobacteria, Planctomycetes, the TM6 group, Firmicutes, and Spirochetes. Several of the phylotypes showed a distinct vertical distribution in the crust, suggesting specific adaptations to the presence or absence of oxygen and light. Archaea were less abundant than Bacteria, their diversity was lower, and the community was less stratified. Detected archaeal groups included organisms affiliated with the Methanosarcinales, the Halobacteriales, and uncultured groups of Euryarchaeota. PMID- 16269780 TI - Detection of airborne Stachybotrys chartarum macrocyclic trichothecene mycotoxins in the indoor environment. AB - The existence of airborne mycotoxins in mold-contaminated buildings has long been hypothesized to be a potential occupant health risk. However, little work has been done to demonstrate the presence of these compounds in such environments. The presence of airborne macrocyclic trichothecene mycotoxins in indoor environments with known Stachybotrys chartarum contamination was therefore investigated. In seven buildings, air was collected using a high-volume liquid impaction bioaerosol sampler (SpinCon PAS 450-10) under static or disturbed conditions. An additional building was sampled using an Andersen GPS-1 PUF sampler modified to separate and collect particulates smaller than conidia. Four control buildings (i.e., no detectable S. chartarum growth or history of water damage) and outdoor air were also tested. Samples were analyzed using a macrocyclic trichothecene-specific enzyme-linked immunosorbent assay (ELISA). ELISA specificity was tested using phosphate-buffered saline extracts of the fungal genera Aspergillus, Chaetomium, Cladosporium, Fusarium, Memnoniella, Penicillium, Rhizopus, and Trichoderma, five Stachybotrys strains, and the indoor air allergens Can f 1, Der p 1, and Fel d 1. For test buildings, the results showed that detectable toxin concentrations increased with the sampling time and short periods of air disturbance. Trichothecene values ranged from <10 to >1,300 pg/m3 of sampled air. The control environments demonstrated statistically significantly (P < 0.001) lower levels of airborne trichothecenes. ELISA specificity experiments demonstrated a high specificity for the trichothecene producing strain of S. chartarum. Our data indicate that airborne macrocyclic trichothecenes can exist in Stachybotrys-contaminated buildings, and this should be taken into consideration in future indoor air quality investigations. PMID- 16269779 TI - Investigation of Oscillatoria spongeliae-dominated bacterial communities in four dictyoceratid sponges. AB - Certain species of marine sponges in the order Dictyoceratida harbor large populations of the cyanobacterial symbiont Oscillatoria spongeliae in the mesohyl (interior) of the sponge. We show that in four of these sponge species (Lamellodysidea herbacea, Lamellodysidea chlorea, Lendenfeldia chondrodes, and Phyllospongia papyracea) from Palau there is a consistent community of alpha proteobacteria in addition to O. spongeliae that fall within the Rhodobacter group based on 16S rRNA gene analysis. Some of the alpha-proteobacteria in Lendenfeldia chondrodes and P. papyracea but not in the Lamellodysidea spp. contained site-specific insertions in the 16S rRNA gene. Reverse transcription PCR experiments demonstrated that the largest insertion found in this study (63 bp) is present in the mature rRNA. Lendenfeldia chondrodes was the only sponge found to have another cyanobacterium in the tissue, a Synechocystis sp. We found that the Synechocystis sp. was present in both the pinacoderm (surface epithelial tissue) and mesohyl, in contrast to O. spongeliae, which was only found in the mesohyl through the use of specific fluorescence in situ hybridization experiments. Of the four sponge species, only P. papyracea was found to contain a significant number of gamma-proteobacteria. These results demonstrate that O. spongeliae-dominated bacterial communities in different sponge species can vary considerably and increase our understanding of the bacterial communities found in marine invertebrates. PMID- 16269781 TI - Anaerobic microbial communities in Lake Pavin, a unique meromictic lake in France. AB - The Bacteria and Archaea from the meromictic Lake Pavin were analyzed in samples collected along a vertical profile in the anoxic monimolimnion and were compared to those in samples from the oxic mixolimnion. Nine targeted 16S rRNA oligonucleotide probes were used to assess the distribution of Bacteria and Archaea and to investigate the in situ occurrence of sulfate-reducing bacteria and methane-producing Archaea involved in the terminal steps of the anaerobic degradation of organic material. The diversity of the complex microbial communities was assessed from the 16S rRNA polymorphisms present in terminal restriction fragment (TRF) depth patterns. The densities of the microbial community increased in the anoxic layer, and Archaea detected with probe ARCH915 represented the largest microbial group in the water column, with a mean Archaea/Eubacteria ratio of 1.5. Terminal restriction fragment length polymorphism (TRFLP) analysis revealed an elevated archaeal and bacterial phylotype richness in anoxic bottom-water samples. The structure of the Archaea community remained rather homogeneous, while TRFLP patterns for the eubacterial community revealed a heterogeneous distribution of eubacterial TRFs. PMID- 16269782 TI - Distribution and diversity of natural product genes in marine and freshwater cyanobacterial cultures and genomes. AB - Natural products are a functionally diverse class of biochemically synthesized compounds, which include antibiotics, toxins, and siderophores. In this paper, we describe both the detection of natural product activities and the sequence identification of gene fragments from two molecular systems that have previously been implicated in natural product production, i.e., nonribosomal peptide synthetases (NRPSs) and modular polyketide synthases (PKSs), in diverse marine and freshwater cyanobacterial cultures. Using degenerate PCR and the sequencing of cloned products, we show that NRPSs and PKSs are common among the cyanobacteria tested. Our molecular data, when combined with genomic searches of finished and progressing cyanobacterial genomes, demonstrate that not all cyanobacteria contain NRPS and PKS genes and that the filamentous and heterocystous cyanobacteria are the richest sources of these genes and the most likely sources of novel natural products within the phylum. In addition to validating the use of degenerate primers for the identification of PKS and NRPS genes in cyanobacteria, this study also defines numerous gene fragments that will be useful as probes for future studies of the synthesis of natural products in cyanobacteria. Phylogenetic analyses of the cyanobacterial NRPS and PKS fragments sequenced in this study, as well as those from the cyanobacterial genome projects, demonstrate that there is remarkable diversity and likely novelty of these genes within the cyanobacteria. These results underscore the potential variety of novel products being produced by these ubiquitous organisms. PMID- 16269783 TI - Identification and molecular characterization of the chromosomal exopolysaccharide biosynthesis gene cluster from Lactococcus lactis subsp. cremoris SMQ-461. AB - The exopolysaccharide (EPS) capsule-forming strain SMQ-461 of Lactococcus lactis subsp. cremoris, isolated from raw milk, produces EPS with an apparent molecular mass of >1.6 x 10(6) Da. The EPS biosynthetic genes are located on the chromosome in a 13.2-kb region consisting of 15 open reading frames. This region is flanked by three IS1077-related tnp genes (L. lactis) at the 5' end and orfY, along with an IS981-related tnp gene, at the 3' end. The eps genes are organized in specific regions involved in regulation, chain length determination, biosynthesis of the repeat unit, polymerization, and export. Three (epsGIK) of the six predicted glycosyltransferase gene products showed low amino acid similarity with known glycosyltransferases. The structure of the repeat unit could thus be different from those known to date for Lactococcus. Reverse transcription-PCR analysis revealed that the eps locus is transcribed as a single mRNA. The function of the eps gene cluster was confirmed by disrupting the priming glycosyltransferase gene (epsD) in Lactococcus cremoris SMQ-461, generating non-EPS-producing reversible mutants. This is the first report of a chromosomal location for EPS genetic elements in Lactococcus cremoris, with novel glycosyltransferases not encountered before in lactic acid bacteria. PMID- 16269784 TI - Real-time PCR quantification of human adenoviruses in urban rivers indicates genome prevalence but low infectivity. AB - Real-time PCR was applied to quantify the abundance of human adenoviruses in two southern California urban rivers, the San Gabriel and Los Angeles. A total of 114 river samples from five different locations were collected over a 1-year period and analyzed for human adenoviruses, along with fecal indicator bacteria and coliphages. Adenoviruses were detected by real-time PCR in approximately 16% of the samples, with concentrations ranging from 10(2) to 10(4) genomes per liter. However, a plaque assay using two human tissue culture cell lines, HEK-293A and A549, yielded negative results, suggesting that adenoviruses detected by real time PCR are likely noninfectious. Enterovirus genome was detected in approximately 7% of the samples by reverse transcription-PCR. Analysis by Spearman's rho rank order correlation showed significant correlations between fecal indicator bacteria and indicator virus (total coliform, fecal coliform, enterococcus, and coliphage values). However, no significant correlations were found between human adenoviruses quantified by real-time PCR and culturable coliphages or fecal indicator bacteria. Kruskal-Wallis chi-square analysis showed significant seasonal variability of all fecal indicator bacteria and coliphages, while no significant variability was observed for adenoviruses or enteroviruses. This study presents the first quantitative measurement of human adenovirus genomes in urban rivers and their statistical relationship to fecal indicator bacteria and coliphages. The uncoupling between high-number genome copies of adenoviruses detected by real-time PCR and the absence of infectivity detected by tissue culture suggests that genome-based detection methods are inadequate for direct assessment of human health risk. PMID- 16269785 TI - Molecular characterization of the nonphotosynthetic partner bacterium in the consortium "Chlorochromatium aggregatum". AB - Phototrophic consortia represent valuable model systems for the study of signal transduction and coevolution between different bacteria. The phototrophic consortium "Chlorochromatium aggregatum" consists of a colorless central rod shaped bacterium surrounded by about 20 green-pigmented epibionts. Although the epibiont was identified as a member of the green sulfur bacteria, and recently isolated and characterized in pure culture, the central colorless bacterium has been identified as a member of the beta-Proteobacteria but so far could not be characterized further. In the present study, "C. aggregatum" was enriched chemotactically, and the 16S rRNA gene sequence of the central bacterium was elucidated. Based on the sequence information, fluorescence in situ hybridization probes targeting four different regions of the 16S rRNA were designed and shown to hybridize exclusively to cells of the central bacterium. Phylogenetic analyses of the 1,437-bp-long sequence revealed that the central bacterium of "C. aggregatum" represents a so far isolated phylogenetic lineage related to Rhodoferax spp., Polaromonas vacuolata, and Variovorax paradoxus within the family Comamonadaceae. The majority of relatives of this lineage are not yet cultured and were found in low-temperature aquatic environments or aquatic environments containing xenobiotica or hydrocarbons. In CsCl-bisbenzimidazole equilibrium density gradients, genomic DNA of the central bacterium of "Chlorochromatium aggregatum" formed a distinct band which could be detected by quantitative PCR using specific primers. Using this method, the G+C content of the central bacterium was determined to be 55.6 mol%. PMID- 16269786 TI - Rhodopseudomonas palustris regulons detected by cross-species analysis of alphaproteobacterial genomes. AB - Rhodopseudomonas palustris, an alpha-proteobacterium, carries out three of the chemical reactions that support life on this planet: the conversion of sunlight to chemical-potential energy; the absorption of carbon dioxide, which it converts to cellular material; and the fixation of atmospheric nitrogen into ammonia. Insight into the transcription-regulatory network that coordinates these processes is fundamental to understanding the biology of this versatile bacterium. With this goal in mind, we predicted regulatory signals genomewide, using a two-step phylogenetic-footprinting and clustering process that we had developed previously. In the first step, 4,963 putative transcription factor binding sites, upstream of 2,044 genes and operons, were identified using cross species Gibbs sampling. Bayesian motif clustering was then employed to group the cross-species motifs into regulons. We have identified 101 putative regulons in R. palustris, including 8 that are of particular interest: a photosynthetic regulon, a flagellar regulon, an organic hydroperoxide resistance regulon, the LexA regulon, and four regulons related to nitrogen metabolism (FixK2, NnrR, NtrC, and sigma54). In some cases, clustering allowed us to assign functions to proteins that previously had been annotated with only putative functions; we have identified RPA0828 as the organic hydroperoxide resistance regulator and RPA1026 as a cell cycle methylase. In addition to predicting regulons, we identified a novel inverted repeat that likely forms a highly conserved stem-loop and that occurs downstream of over 100 genes. PMID- 16269787 TI - Global transcriptional profiling of Shewanella oneidensis MR-1 during Cr(VI) and U(VI) reduction. AB - Whole-genome DNA microarrays were used to examine the gene expression profile of Shewanella oneidensis MR-1 during U(VI) and Cr(VI) reduction. The same control, cells pregrown with nitrate and incubated with no electron acceptor, was used for the two time points considered and for both metals. U(VI)-reducing conditions resulted in the upregulation (> or = 3-fold) of 121 genes, while 83 genes were upregulated under Cr(VI)-reducing conditions. A large fraction of the genes upregulated [34% for U(VI) and 29% for Cr(VI)] encode hypothetical proteins of unknown function. Genes encoding proteins known to reduce alternative electron acceptors [fumarate, dimethyl sulfoxide, Mn(IV), or soluble Fe(III)] were upregulated under both U(VI)- and Cr(VI)-reducing conditions. The involvement of these upregulated genes in the reduction of U(VI) and Cr(VI) was tested using mutants lacking one or several of the gene products. Mutant testing confirmed the involvement of several genes in the reduction of both metals: mtrA, mtrB, mtrC, and menC, all of which are involved in Fe(III) citrate reduction by MR-1. Genes encoding efflux pumps were upregulated under Cr(VI)- but not under U(VI)-reducing conditions. Genes encoding proteins associated with general (e.g., groL and dnaJ) and membrane (e.g., pspBC) stress were also upregulated, particularly under U(VI) reducing conditions, pointing to membrane damage by the solid-phase reduced U(IV) and Cr(III) and/or the direct effect of the oxidized forms of the metals. This study sheds light on the multifaceted response of MR-1 to U(VI) and Cr(VI) under anaerobic conditions and suggests that the same electron transport pathway can be used for more than one electron acceptor. PMID- 16269788 TI - Proof that Burkholderia strains form effective symbioses with legumes: a study of novel Mimosa-nodulating strains from South America. AB - Twenty Mimosa-nodulating bacterial strains from Brazil and Venezuela, together with eight reference Mimosa-nodulating rhizobial strains and two other beta rhizobial strains, were examined by amplified rRNA gene restriction analysis. They fell into 16 patterns and formed a single cluster together with the known beta-rhizobia, Burkholderia caribensis, Burkholderia phymatum, and Burkholderia tuberum. The 16S rRNA gene sequences of 15 of the 20 strains were determined, and all were shown to belong to the genus Burkholderia; four distinct clusters could be discerned, with strains isolated from the same host species usually clustering very closely. Five of the strains (MAP3-5, Br3407, Br3454, Br3461, and Br3469) were selected for further studies of the symbiosis-related genes nodA, the NodD dependent regulatory consensus sequences (nod box), and nifH. The nodA and nifH sequences were very close to each other and to those of B. phymatum STM815, B. caribensis TJ182, and Cupriavidus taiwanensis LMG19424 but were relatively distant from those of B. tuberum STM678. In addition to nodulating their original hosts, all five strains could also nodulate other Mimosa spp., and all produced nodules on Mimosa pudica that had nitrogenase (acetylene reduction) activities and structures typical of effective N2-fixing symbioses. Finally, both wild-type and green fluorescent protein-expressing transconjugant strains of Br3461 and MAP3-5 produced N2-fixing nodules on their original hosts, Mimosa bimucronata (Br3461) and Mimosa pigra (MAP3-5), and hence this confirms strongly that Burkholderia strains can form effective symbioses with legumes. PMID- 16269789 TI - First genome data from uncultured upland soil cluster alpha methanotrophs provide further evidence for a close phylogenetic relationship to Methylocapsa acidiphila B2 and for high-affinity methanotrophy involving particulate methane monooxygenase. AB - Members of upland soil cluster alpha (USC alpha) are assumed to be methanotrophic bacteria (MB) adapted to the trace level of atmospheric methane. So far, these MB have eluded all cultivation attempts. While the 16S rRNA phylogeny of USC alpha members is still not known, phylogenies constructed for the active-site polypeptide (encoded by pmoA) of particulate methane monooxygenase (pMMO) placed USC alpha next to the alphaproteobacterial Methylocapsa acidiphila B2. To assess whether the pmoA tree reflects the evolutionary identity of USC alpha, a 42-kb genomic contig of a USC alpha representative was obtained from acidic forest soil by screening a metagenomic fosmid library of 250,000 clones using pmoA-targeted PCR. For comparison, a 101-kb genomic contig from M. acidiphila was analyzed, including the pmo operon. The following three lines of evidence confirmed a close phylogenetic relationship between USC alpha and M. acidiphila: (i) tetranucleotide frequency patterns of 5-kb genomic subfragments, (ii) annotation and comparative analysis of the genomic fragments against all completely sequenced genomes available in public domain databases, and (iii) three single gene phylogenies constructed using the deduced amino acid sequences of a putative prephenate dehydratase, a staphylococcal-like nuclease, and a putative zinc metalloprotease. A comparative analysis of the pmo operons of USC alpha and M. acidiphila corroborated previous reports that both the pmo operon structure and the predicted secondary structure of deduced pMMO are highly conserved among all MB. PMID- 16269790 TI - Colonization of mucin by human intestinal bacteria and establishment of biofilm communities in a two-stage continuous culture system. AB - The human large intestine is covered with a protective mucus coating, which is heavily colonized by complex bacterial populations that are distinct from those in the gut lumen. Little is known of the composition and metabolic activities of these biofilms, although they are likely to play an important role in mucus breakdown. The aims of this study were to determine how intestinal bacteria colonize mucus and to study physiologic and enzymatic factors involved in the destruction of this glycoprotein. Colonization of mucin gels by fecal bacteria was studied in vitro, using a two-stage continuous culture system, simulating conditions of nutrient availability and limitation characteristic of the proximal (vessel 1) and distal (vessel 2) colon. The establishment of bacterial communities in mucin gels was investigated by selective culture methods, scanning electron microscopy, and confocal laser scanning microscopy, in association with fluorescently labeled 16S rRNA oligonucleotide probes. Gel samples were also taken for analysis of mucin-degrading enzymes and measurements of residual mucin sugars. Mucin gels were rapidly colonized by heterogeneous bacterial populations, especially members of the Bacteroides fragilis group, enterobacteria, and clostridia. Intestinal bacterial populations growing on mucin surfaces were shown to be phylogenetically and metabolically distinct from their planktonic counterparts. PMID- 16269791 TI - Diversity, localization, and physiological properties of filamentous microbes belonging to Chloroflexi subphylum I in mesophilic and thermophilic methanogenic sludge granules. AB - We previously reported that the thermophilic filamentous anaerobe Anaerolinea thermophila, which is the first cultured representative of subphylum I of the bacterial phylum Chloroflexi, not only was one of the predominant constituents of thermophilic sludge granules but also was a causative agent of filamentous sludge bulking in a thermophilic (55 degrees C) upflow anaerobic sludge blanket (UASB) reactor in which high-strength organic wastewater was treated (Y. Sekiguchi, H. Takahashi, Y. Kamagata, A. Ohashi, and H. Harada, Appl. Environ. Microbiol. 67:5740-5749, 2001). To further elucidate the ecology and function of Anaerolinea type filamentous microbes in UASB sludge granules, we surveyed the diversity, distribution, and physiological properties of Chloroflexi subphylum I microbes residing in UASB granules. Five different types of mesophilic and thermophilic UASB sludge were used to analyze the Chloroflexi subphylum I populations. 16S rRNA gene cloning-based analyses using a 16S rRNA gene-targeted Chloroflexi specific PCR primer set revealed that all clonal sequences were affiliated with the Chloroflexi subphylum I group and that a number of different phylotypes were present in each clone library, suggesting the ubiquity and vast genetic diversity of these populations in UASB sludge granules. Subsequent fluorescence in situ hybridization (FISH) of the three different types of mesophilic sludge granules using a Chloroflexi-specific probe suggested that all probe-reactive cells had a filamentous morphology and were widely distributed within the sludge granules. The FISH observations also indicated that the Chloroflexi subphylum I bacteria were not always the predominant populations within mesophilic sludge granules, in contrast to thermophilic sludge granules. We isolated two mesophilic strains and one thermophilic strain belonging to the Chloroflexi subphylum I group. The physiological properties of these isolates suggested that these populations may contribute to the degradation of carbohydrates and other cellular components, such as amino acids, in the bioreactors. PMID- 16269792 TI - Design, validation, and application of a seven-strain Staphylococcus aureus PCR product microarray for comparative genomics. AB - Bacterial comparative genomics has been revolutionized by microarrays, but the power of any microarray is dependent on the number and diversity of gene reporters it contains. Staphylococcus aureus is an important human pathogen causing a wide range of invasive and toxin-mediated diseases, and more than 20% of the genome of any isolate consists of variable genes. Seven whole-genome sequences of S. aureus are available, and we exploited this rare opportunity to design, build, and validate a comprehensive, nonredundant PCR product microarray carrying reporters that represent every predicted open reading frame (3,623 probes). Such a comprehensive microarray necessitated a novel design strategy. Validation with the seven sequenced strains showed correct identification of 93.9% of genes present or absent/divergent but was dependent on the method of analysis chosen. Microarray data were highly reproducible, reducing the need for many replicate slides. Interpretation of microarray data was enhanced by focusing on the major areas of variation--the presence or absence of mobile genetic elements (MGEs). We compiled "composite genomes" of every individual MGE and visualized their distribution. This allowed the sensitive discrimination of related isolates, including the first clear description of how isolates of the same clone of epidemic methicillin-resistant S. aureus differ substantially in their carriage of MGEs. These MGEs carry virulence and resistance genes, suggesting differences in pathogenic potential. The novel methods of design and interpretation of data generated from this microarray will enable further studies of S. aureus evolution, epidemiology, and pathogenesis. PMID- 16269793 TI - Isolation, sequence analysis, and comparison of two plasmids (28 and 29 kilobases) from the biomining bacterium Leptospirillum ferrooxidans ATCC 49879. AB - Two plasmids, of 28,878 bp and 28,012 bp, were isolated from Leptospirillum ferrooxidans ATCC 49879. Altogether, a total of 67 open reading frames (ORFs) were identified on both plasmids, of which 32 had predicted products with high homology to proteins of known function, while 11 ORFs had predicted products with homology to previously identified proteins of unknown function. Twenty-four ORFs had products with no homologues in the GenBank/NCBI database. An analysis of the ORFs and other features of the two plasmids, the first to be isolated from a bacterium of the genus Leptospirillum, is presented. PMID- 16269795 TI - Application of measurements of transepithelial electrical resistance of intestinal epithelial cell monolayers to evaluate probiotic activity. AB - Among five potentially probiotic lactobacilli investigated, Lactobacillus plantarum MF1298 and Lactobacillus salivarius DC5 showed the highest increase in the transepithelial electrical resistance (TER) of polarized monolayers of Caco-2 cells, and this increase was shown to be dose dependent. Furthermore, preincubation with MF1298 attenuated a decrease in TER induced by Listeria monocytogenes. PMID- 16269794 TI - Distribution, localization, and phylogeny of abundant populations of Crenarchaeota in anaerobic granular sludge. AB - Eight anaerobic granular sludges were surveyed for Crenarchaeota using rRNA gene cloning. Microbial arrangement and substrate uptake patterns were elucidated by fluorescent in situ hybridization and beta imaging. Group 1.3 Crenarchaeota represented up to 50% of Archaea and 25% of the total microbiota in five sludges. Crenarchaeota were localized in close association with methanogenic Archaea. PMID- 16269796 TI - Sulfate-reducing bacteria in floating macrophyte rhizospheres from an Amazonian floodplain lake in Bolivia and their association with Hg methylation. AB - Five subgroups of sulfate-reducing bacteria (SRB) were detected by PCR in three macrophyte rhizospheres (Polygonum densiflorum, Hymenachne donacifolia, and Ludwigia helminthorriza) and three subgroups in Eichhornia crassipes from La Granja, a floodplain lake from the upper Madeira basin. The SRB community varied according to the macrophyte species but with different degrees of association with their roots. The rhizosphere of the C4 plant Polygonum densiflorum had higher frequencies of SRB subgroups as well as higher mercury methylation potentials (27.5 to 36.1%) and carbon (16.06 +/- 5.40%), nitrogen (2.03 +/- 0.64%), Hg (94.50 +/- 6.86 ng Hg g(-1)), and methylmercury (8.25 +/- 1.45 ng Hg g(-1)) contents than the rhizosphere of the C3 plant Eichhornia crassipes. Mercury methylation in Polygonum densiflorum and Eichhornia crassipes was reduced when SRB metabolism was inhibited by sodium molybdate. PMID- 16269797 TI - Symbiotic hydrogenase activity in Bradyrhizobium sp. (Vigna) increases nitrogen content in Vigna unguiculata plants. AB - Bradyrhizobium sp. (Lupinus) and Bradyrhizobium sp. (Vigna) mutants in which hydrogenase (hup) activity was affected were constructed and analyzed. Vigna unguiculata plants inoculated with the Bradyrhizobium sp. (Vigna) hup mutant showed reduced nitrogenase activity and also a significant decrease in nitrogen content, suggesting a relevant contribution of hydrogenase activity to plant yield. PMID- 16269798 TI - On-probe sample pretreatment for direct analysis of lipids in gram-positive bacterial cells by matrix-assisted laser desorption ionization mass spectrometry. AB - On-probe sample pretreatment using trifluoroacetic acid as an additional reagent enabled the direct detection of phospholipids in whole bacteria by means of matrix-assisted laser desorption ionization mass spectrometry for not only gram negative organisms but also gram-positive ones with a thicker peptidoglycan layer. PMID- 16269799 TI - Construction of an alpha toxin gene knockout mutant of Clostridium perfringens type A by use of a mobile group II intron. AB - In developing Clostridium perfringens as a safe vaccine vector, the alpha toxin gene (plc) in the bacterial chromosome must be permanently inactivated. Disrupting genes in C. perfringens by traditional mutagenesis methods is very difficult. Therefore, we developed a new strategy using group II intron-based Target-Tron technology to inactivate the plc gene in C. perfringens ATCC 3624. Western blot analysis showed no production of alpha toxin protein in the culture supernatant of the plc mutant. Advantages of this technology, such as site specificity, relatively high frequency of insertion, and introduction of no antibiotic resistance genes into the chromosome, could facilitate construction of other C. perfringens mutants. PMID- 16269800 TI - Cutinase-like enzyme from the yeast Cryptococcus sp. strain S-2 hydrolyzes polylactic acid and other biodegradable plastics. AB - A purified lipase from the yeast Cryptococcus sp. strain S-2 exhibited remote homology to proteins belonging to the cutinase family rather than to lipases. This enzyme could effectively degrade the high-molecular-weight compound polylactic acid, as well as other biodegradable plastics, including polybutylene succinate, poly (epsilon-caprolactone), and poly(3-hydroxybutyrate). PMID- 16269801 TI - Strain- and genotype-specific differences in virulence of Paenibacillus larvae subsp. larvae, a bacterial pathogen causing American foulbrood disease in honeybees. AB - Virulence variations of Paenibacillus larvae subsp. larvae, the causative agent of American foulbrood disease of honeybees, were investigated by analysis of 16 field isolates of this pathogen, belonging to three previously characterized genotypes, as well as the type strain (ATCC 9545) of P. larvae subsp. larvae, with exposure bioassays. We demonstrated that the strain-specific 50% lethal concentrations varied within an order of magnitude and that differences in amount of time for the pathogen to kill 100% of the infected hosts (LT100) correlated with genotype. One genotype killed rather quickly, with a mean LT100 of 7.8 +/- 1.7 days postinfection, while the other genotypes acted more slowly, with mean LT100s of 11.2 +/- 0.8 and 11.6 +/- 0.6 days postinfection. PMID- 16269803 TI - Putative exposed aromatic and hydroxyl residues on the surface of the N-terminal domains of Chi1 from Aeromonas caviae CB101 are essential for chitin binding and hydrolysis. AB - Chitinase Chi1 of Aeromonas caviae CB101 possesses chitin binding sites at both its N and C termini. Four putative exposed residues aligned in a line on the surface of the N-terminal domains of Chi1 were found to contribute to the enzyme chitin binding and hydrolysis via site-directed mutagenesis. Also, it was found that Chi1 requires the cooperation of the N- and C-terminal domains to bind fully with crystalline and colloidal chitin. PMID- 16269802 TI - Evidence for horizontal transfer of 1-aminocyclopropane-1-carboxylate deaminase genes. AB - PCR was used to rapidly identify and isolate 1-aminocyclopropane-1-carboxylate (ACC) deaminase genes from bacteria. The Shimodaira-Hasegawa test was used to assess whether phylogenetically anomalous gene placements suggestive of horizontal gene transfer (HGT) were significantly favored over vertical transmission. The best maximum likelihood (ML) ACC deaminase tree was significantly more likely than four alternative ML trees, suggesting HGT. PMID- 16269804 TI - Occurrence of Listeria spp. in effluents of French urban wastewater treatment plants. AB - Listeria spp. were found in most treated waters (84.4%) and raw sludge (89.2%) of six French urban wastewater treatment plants and one composting facility, examined monthly over a 1-year period. Most strains belonged to Listeria monocytogenes, serotypes 4b/4e being predominant. Sludge composting and liming reduced or prevented Listeria contamination. PMID- 16269805 TI - Modeling bacteriocin resistance and inactivation of Listeria innocua LMG 13568 by Lactobacillus sakei CTC 494 under sausage fermentation conditions. AB - In mixed cultures, bacteriocin production by the sausage isolate Lactobacillus sakei CTC 494 rapidly inactivated sensitive Listeria innocua LMG 13568 cells, even at low bacteriocin activity levels. A small fraction of the listerial population was bacteriocin resistant. However, sausage fermentation conditions inhibited regrowth of resistant cells. PMID- 16269806 TI - Factors influencing the ability of Listeria monocytogenes to pass through a membrane filter by active infiltration. AB - Listeria monocytogenes infiltrated the reticulate structure of a membrane filter and passed through a filter with pore sizes of 0.45 microm and 0.2 microm in 6 to 24 h and 5 to 6 days, respectively. Flagellar motility and expansive pressure generated by the growing bacterial population were indicated as the driving forces of infiltration. PMID- 16269807 TI - Impact of bacterial NO3(-) transport on sediment biogeochemistry. AB - Experiments demonstrated that Beggiatoa could induce a H2S-depleted suboxic zone of more than 10 mm in marine sediments and cause a divergence in sediment NO3(-) reduction from denitrification to dissimilatory NO3(-) reduction to ammonium. pH, O2, and H2S profiles indicated that the bacteria oxidized H2S with NO3(-) and transported S0 to the sediment surface for aerobic oxidation. PMID- 16269808 TI - Representational difference analysis and real-time PCR for strain-specific quantification of Lactobacillus sobrius sp. nov. AB - Lactobacillus sobrius sp. nov., which was recently isolated from the intestine of weaning piglets, has potential probiotic properties. To follow the fate of L. sobrius strain 001T in dietary interventions, a novel and strain-specific quantitative detection procedure was developed. This procedure was based on the isolation of specific genomic fragments from the type strain by representational difference analysis and their detection by real-time PCR. The described strain specific quantification approach may be used in studies aimed at tracking bacterial strains added to specific environments. PMID- 16269809 TI - Heterogeneity of putative surface layer proteins in Lactobacillus helveticus. AB - The S-layer-encoding genes of 21 Lactobacillus helveticus strains were characterized. Phylogenetic analysis based on the identified S-layer genes revealed two main clusters, one which includes a sequence similar to that of the slpH1 gene of L. helveticus CNRZ 892 and a second cluster which includes genes similar to that of prtY. These results were further confirmed by Southern blot hybridization. This study demonstrates S-layer gene variability in the species L. helveticus. PMID- 16269810 TI - Bactericidal actions of a silver ion solution on Escherichia coli, studied by energy-filtering transmission electron microscopy and proteomic analysis. AB - Bactericidal actions of the silver ion on Escherichia coli as a model microorganism were studied using energy-filtering transmission electron microscopy (EFTEM), two-dimensional electrophoresis (2-DE), and matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF MS). EFTEM observations demonstrated that the silver ion readily infiltrates the interior of E. coli, contrary to the early hypothesis that it resides initially in the cell membrane area. Furthermore, 2-DE and MALDI-TOF MS indicated that the expression of a ribosomal subunit protein as well as that of some other enzymes and proteins is affected by the silver ion. The present results demonstrate for the first time that one of the major bactericidal functions of the silver ion is its interaction with the ribosome and the ensuing inhibition in expression of the enzymes and proteins essential to ATP production. PMID- 16269811 TI - Discrimination between Francisella tularensis and Francisella-like endosymbionts when screening ticks by PCR. AB - The presence of Francisella-like endosymbionts in tick species known to transmit tularemia poses a potential diagnostic problem for laboratories that screen tick samples by PCR for Francisella tularensis. Tick samples initially considered positive for F. tularensis based on standard 16S rRNA gene PCR were found to be positive only for Francisella-like endosymbionts using a multitarget F. tularensis TaqMan assay (ISFtu2, tul4, and iglC) and 16S rRNA gene sequencing. Specificity of PCR-based diagnostics for F. tularensis should be carefully evaluated with appropriate specimen types prior to diagnostic use. PMID- 16269812 TI - Metagenomic analysis of the microbial community at Zodletone Spring (Oklahoma): insights into the genome of a member of the novel candidate division OD1. AB - A metagenomic library was constructed from the anaerobic sediments of a mesophilic sulfur spring. Thirty-five bacterial 16S rRNA gene-containing clones were identified in this library. Analysis of a genomic fragment belonging to candidate division OD1 provided useful insights into the physiology and biochemistry of this novel, yet-uncultured candidate division. PMID- 16269813 TI - Rhizobium leguminosarum biovar viciae symbiotic hydrogenase activity and processing are limited by the level of nickel in agricultural soils. AB - Analysis of levels of hydrogenase processing and activity in Rhizobium leguminosarum biovar viciae bacteroids from pea (Pisum sativum) plants showed that the oxidation of nitrogenase-evolved hydrogen is limited by the availability of nickel in agricultural soils. This limitation was overcome by using an inoculant strain engineered for higher hydrogenase expression. PMID- 16269814 TI - Creating new genes by plasmid recombination in Escherichia coli and Bacillus subtilis. AB - Gene shuffling is a way of creating proteins with interesting new characteristics, starting from diverged sequences. We tested an alternative to gene shuffling based on plasmid recombination and found that Bacillus subtilis efficiently recombines sequences with 4% divergence, and Escherichia coli mutS is more appropriate for sequences with 22% divergence. PMID- 16269815 TI - When coupled to natural transformation in Acinetobacter sp. strain ADP1, PCR mutagenesis is made less random by mismatch repair. AB - Random PCR mutagenesis is a powerful tool for structure-function analysis of targeted proteins, especially when coupled with DNA integration through natural transformation followed by selection for loss of function. The technique has been applied successfully to structure-function analysis of transcriptional regulators, enzymes, and transporters in Acinetobacter sp. strain ADP1. However, the mismatch repair system prevents the full spectrum of nucleotide substitutions that may be selected at the level of protein function from being recovered. This barrier may be overcome by introducing PCR-mutagenized genes into strains in which the corresponding genes have been deleted. PMID- 16269816 TI - Bacteriocin (mutacin) production by Streptococcus mutans genome sequence reference strain UA159: elucidation of the antimicrobial repertoire by genetic dissection. AB - Streptococcus mutans UA159, the genome sequence reference strain, exhibits nonlantibiotic mutacin activity. In this study, bioinformatic and mutational analyses were employed to demonstrate that the antimicrobial repertoire of strain UA159 includes mutacin IV (specified by the nlm locus) and a newly identified bacteriocin, mutacin V (encoded by SMU.1914c). PMID- 16269817 TI - Comparison of the levels of heat resistance of wild-type, cpe knockout, and cpe plasmid-cured Clostridium perfringens type A strains. AB - An enterotoxin (cpe) plasmid was cured from a Clostridium perfringens non-food borne gastrointestinal disease (NFBGID) isolate, and the heat resistance levels of wild-type, cpe knockout, and cpe plasmid-cured strains were compared. Our results demonstrated that (i) wild-type cpe has no influence in mediating high level heat resistance in C. perfringens and (ii) the cpe plasmid does not confer heat sensitivity on NFBGID isolates. PMID- 16269818 TI - Identification of pilus-like structures and genes in Microcystis aeruginosa PCC7806. AB - Four putative type IV pilus genes from the toxic, naturally transformable Microcystis aeruginosa PCC7806 were identified. Three of these genes were clustered in an arrangement which is identical to that from other cyanobacterial genomes. Type IV pilus-like appendages were also observed by electron microscopy. PMID- 16269819 TI - Trichosporon species isolated from guano samples obtained from bat-inhabited caves in Japan. AB - Yeasts from caves have rarely been examined. We examined yeasts collected from bat guano samples from 20 bat-inhabited limestone and volcanic caves located in 11 prefectures in Japan. Of approximately 700 yeast-like colonies, nine Trichosporon species were recovered from 15 caves. Two of these were known species, and the remaining seven are potentially novel species, based on molecular phylogenetic analyses. In addition to Trichosporon species, identifiable strains of eight ascomycetous yeasts and one basidiomycetous yeast were recovered at frequencies of 5 to 35%. Our findings suggest that Trichosporon spp. are the major yeast species in bat guano in Japan and that bat guano is a potentially rich source of previously undescribed yeast species. PMID- 16269820 TI - In vitro non-viral gene delivery with nanofibrous scaffolds. AB - Extracellular and intracellular barriers typically prevent non-viral gene vectors from having an effective transfection efficiency. Formulation of a gene delivery vehicle that can overcome the barriers is a key step for successful tissue regeneration. We have developed a novel core-shelled DNA nanoparticle by invoking solvent-induced condensation of plasmid DNA (beta-galactosidase or GFP) in a solvent mixture [94% N,N-dimethylformamide (DMF) + 6% 1x TE buffer] and subsequent encapsulation of the condensed DNA globule in a triblock copolymer, polylactide-poly(ethylene glycol)-polylactide (L8E78L8), in the same solvent environment. The polylactide shell protects the encapsulated DNA from degradation during electrospinning of a mixture of encapsulated DNA nanoparticles and biodegradable PLGA (a random copolymer of lactide and glycolide) to form a nanofibrous non-woven scaffold using the same solution mixture. The bioactive plasmid DNA can then be released in an intact form from the scaffold with a controlled release rate and transfect cells in vitro. PMID- 16269821 TI - Binding of the Bacillus subtilis LexA protein to the SOS operator. AB - The Bacillus subtilis LexA protein represses the SOS response to DNA damage by binding as a dimer to the consensus operator sequence 5'-CGAACN(4)GTTCG-3'. To characterize the requirements for LexA binding to SOS operators, we determined the operator bases needed for site-specific binding as well as the LexA amino acids required for operator recognition. Using mobility shift assays to determine equilibrium constants for B.subtilis LexA binding to recA operator mutants, we found that several single base substitutions within the 14 bp recA operator sequence destabilized binding enough to abolish site-specific binding. Our results show that the AT base pairs at the third and fourth positions from the 5' end of a 7 bp half-site are essential and that the preferred binding site for a LexA dimer is 5'-CGAACATATGTTCG-3'. Binding studies with LexA mutants, in which the solvent accessible amino acid residues in the putative DNA binding domain were mutated, indicate that Arg-49 and His-46 are essential for binding and that Lys-53 and Ala-48 are also involved in operator recognition. Guided by our mutational analyses as well as hydroxyl radical footprinting studies of the dinC and recA operators we docked a computer model of B.subtilis LexA on the preferred operator sequence in silico. Our model suggests that binding by a LexA dimer involves bending of the DNA helix within the internal 4 bp of the operator. PMID- 16269822 TI - Tracking of microinjected DNA in live cells reveals the intracellular behavior and elimination of extrachromosomal genetic material. AB - We here addressed the basic question, how does extrachromosomal DNA behave when it is placed in the nuclear or the cytoplasmic environment and how is it eliminated? To do this, we tracked microinjected DNA molecules in live cells. In the cytoplasm, the diffusion of microinjected DNA was inhibited in a size- and linearity-dependent manner, probably by the intermediate filament. This was followed by the rapid disappearance of the DNA fluorescent signal. In the nucleus, the diffusion was also dependent on the size of the molecule and was accompanied by the aggregation of the DNA. The aggregation may be due to a putative DNA-binding molecule, whose level is high during the G1 phase. Surprisingly, the injected DNA could move across the nuclear membrane and appeared in the cytoplasm, which suggests the presence of a transport system. The intracytoplasmic behavior and the elimination of such DNA was obviously different from the DNA that was directly injected at the cytoplasm. The DNA remaining in the nucleus appeared to be stable and persisted in the nucleus or, after cell division, in the cytoplasm, for more than one cell cycle. These findings provide a novel and basic understanding of the behavior and elimination of a wide variety of extrachromosomal genetic material. PMID- 16269823 TI - Duplication processes in Saccharomyces cerevisiae haploid strains. AB - Duplication is thought to be one of the main processes providing a substrate on which the effects of evolution are visible. The mechanisms underlying this chromosomal rearrangement were investigated here in the yeast Saccharomyces cerevisiae. Spontaneous revertants containing a duplication event were selected and analyzed. In addition to the single gene duplication described in a previous study, we demonstrated here that direct tandem duplicated regions ranging from 5 to 90 kb in size can also occur spontaneously. To further investigate the mechanisms in the duplication events, we examined whether homologous recombination contributes to these processes. The results obtained show that the mechanisms involved in segmental duplication are RAD52-independent, contrary to those involved in single gene duplication. Moreover, this study shows that the duplication of a given gene can occur in S.cerevisiae haploid strains via at least two ways: single gene or segmental duplication. PMID- 16269824 TI - FOB1 affects DNA topoisomerase I in vivo cleavages in the enhancer region of the Saccharomyces cerevisiae ribosomal DNA locus. AB - In Saccharomyces cerevisiae the FOB1 gene affects replication fork blocking activity at the replication fork block (RFB) sequences and promotes recombination events within the rDNA cluster. Using in vivo footprinting assays we mapped two in vivo Fob1p-binding sites, RFB1 and RFB3, located in the rDNA enhancer region and coincident with those previously reported to be in vitro binding sites. We previously provided evidences that DNA topoisomerase I is able to cleave two sites within this region. The results reported in this paper, indicate that the DNA topoisomerase I cleavage specific activity at the enhancer region is affected by the presence of Fob1p and independent of replication and transcription activities. We thus hypothesize that the binding to DNA of Fob1p itself may be the cause of the DNA topoisomerase I activity in the rDNA enhancer. PMID- 16269825 TI - A role for FXR and human FGF-19 in the repression of paraoxonase-1 gene expression by bile acids. AB - Paraoxonase-1 (PON1), an enzyme that metabolizes organophosphate insecticides, is secreted by the liver and transported in the blood complexed to HDL. In humans and mice, low plasma levels of PON1 have also been linked to the development of atherosclerosis. We previously reported that hepatic Pon1 expression was decreased when C57BL/6J mice were fed a high-fat, high-cholesterol diet supplemented with cholic acid (CA). In the current study, we used wild-type and farnesoid X receptor (FXR) null mice to demonstrate that this repression is dependent upon CA and FXR. PON1 mRNA levels were also repressed when HepG2 cells, derived from a human hepatoma, were incubated with natural or highly specific synthetic FXR agonists. In contrast, fibroblast growth factor-19 (FGF-19) mRNA levels were greatly induced by these same FXR agonists. Furthermore, treatment of HepG2 cells with recombinant human FGF-19 significantly decreased PON1 mRNA levels. Finally, deletion studies revealed that the proximal -230 to -96 bp region of the PON1 promoter contains regulatory element(s) necessary for promoter activity and bile acid repression. These data demonstrate that human PON1 expression is repressed by bile acids through the actions of FXR and FGF-19. PMID- 16269827 TI - The Bioinformatics Open Access option. PMID- 16269826 TI - Further characterization of mammalian ceramide kinase: substrate delivery and (stereo)specificity, tissue distribution, and subcellular localization studies. AB - Recombinant human ceramide kinase (HsCERK) was analyzed with regard to dependence on divalent cations and to substrate delivery, spectrum, specificity, and stereoselectivity. Depending on the chain length of the ceramide, either albumin for short-chain ceramide or a mixed micellar form (octylglucoside/cardiolipin) for long-chain ceramide was preferred for the substrate delivery, the former resulting in higher activities. Bacterially expressed HsCERK was highly dependent on Mg2+ ions, much less on Ca2+ ions. A clear preference for the d-erythro isomer was seen. Various N-acylated amino alcohols were no substrate, but N-hexanoyl-1-O hexadecyl-2-desoxy-2-amino-sn-glycerol and N-tetradecanoyl-2S-amino-1-butanol were phosphorylated, suggesting that the secondary hydroxy group is not required for recognition. The properties of HsCERK, expressed in CHO cells, were similar to those of the bacterially expressed protein, including the Mg2+ dependence. In mouse, the highest activities were found in testis and cerebellum, and upon subcellular fractionation the activity was recovered mainly in the microsomal fraction. This fits with the plasma membrane localization in CHO cells, which was mediated by the N-terminal putative pleckstrin domain. No evidence for phosphorylation of ceramide by the recently described multiple lipid kinase was found. The latter kinase is localized in the mitochondria, but no firm conclusions with regard to its substrate could be drawn. PMID- 16269828 TI - Neonatal survival and beyond. PMID- 16269829 TI - Cost of neonatal intensive care in a tertiary care center. AB - BACKGROUND: The number of neonatal intensive care units (NICUs) in India has increased substantially over the last decade; yet many more are required. There is limited information on the actual costs of setting up and running an NICU in India. OBJECTIVE: Systematic and comprehensive calculation and analysis of the costs of neonatal intensive care in a tertiary care teaching hospital. METHODS: The costs were compiled by studying the detailed records of various hospital departments and prospectively documenting the costs of drugs, consumables and investigations for a representative group of 30 babies. RESULTS: The total cost of establishing a 16 bed level III tertiary care NICU was Rs 3.78 crore (Rs. 37.8 million, USdollar 860,000) (2003). Equipment cost formed two-thirds of the establishment cost. The running cost of NICU care per patient per day was Rs 5450 (USdollar 125). NICU and ancillary personnel salary comprised the largest proportion of the running costs. The average total cost of care for a baby less than 1000 grams was Rs. 168000 (USdollar 3800), Rs. 88300 (USdollar 2000) for babies 1000 g to 1250 g. and Rs. 41700 (USdollar 950) for those between 1250 to 1500 g. The family had to bear only 25 percent; rest was subsidized. CONCLUSIONS: Equipment and personnel salary form the biggest proportion of establishment and running costs. The costs of treatment for a baby in NICU should be seen in context with costs of other types of health care and the number of useful life years gained. PMID- 16269830 TI - Transcutaneous absorption of topically massaged oil in neonates. AB - OBJECTIVE: To study the transcutaneous absorption of traditionally massaged oil in newborns and to specifically compare the effects of (i) essential fatty acid (EFA) rich - safflower oil and (ii) saturated fat rich coconut oil, on fatty acid profiles of massaged babies. DESIGN: A short term randomised controlled study. SETTING: Tertiary care NICU of a large teaching hospital and a research laboratory of a University complex. METHODS: 120 study babies were randomly assigned to three oil groups (i) safflower oil (n = 40) (ii) coconut oil (n = 40) and (iii) no oil controls (n = 40). In each group, babies were selected in three subsets as per their gestational ages viz (a) less than 34 weeks, (b) 34-37 weeks, (c) greater than 37 weeks. 5 mL of the designated oil was massaged four times a day for five days under controlled conditions of temperature and feeding. Pre and post oil massage samples of blood were analysed for triglycerides and fatty acid profiles using gas chromatography. RESULTS: Post oil triglyceride values were significantly raised in both the oil groups and also in controls. However, the quantum of rise was significantly higher in oil groups as compared to controls. Fatty acid profiles (gas chromatography) showed significant rise in EFAs (linolenic acid and arachidonic acid) in safflower oil group and saturated fats in coconut oil group. Changes were more evident in term babies. There were no side effects associated with the massage. CONCLUSIONS: This study shows that topically applied oil can be absorbed in neonates and is probably available for nutritional purposes. The fatty acid constituents of the oil can influence the changes in the fatty acid profiles of the massaged babies. PMID- 16269831 TI - Preservatives in DPT vaccine. PMID- 16269833 TI - DTP Hib combination. PMID- 16269835 TI - Study of drugs in Indian children. PMID- 16269837 TI - Eligibility of children for vitamin A supplementation program. PMID- 16269838 TI - Myocardial ischemia induced by nebulized fenoterol for severe childhood asthma. AB - We examined for myocardial ischemia induced by continuous inhalation of fenoterol in children with severe acute asthma. Thirty children with severe acute asthma were evaluated for signs of myocardial ischemia when treated with 0.5 mg kg dose (maximum 15 mg) of inhaled fenoterol for one hour. The heart rate was measured before and after inhalation. Cardiac enzymes (creatine kinase, creatine kinase MB fraction and troponin levels) were measured at admission and 12 hours later. An EKG was recorded before inhalation was started and immediately after its completion to detect the presence of any evidence of myocardial ischemia. All patients developed significant increase in heart rate. Six patients showed EKG changes compatible with myocardial ischemia, despite normal enzyme levels. Patients with severe acute asthma show tachycardia and may show EKG changes of myocardial ischemia. PMID- 16269839 TI - Pyogenic meningitis in Ilesa, Nigeria. AB - This study was conducted in 124 children with meningitis to review the etiology, antimicrobial susceptibility and outcome of disease in a Nigerian tertiary health facility. Of these, 97 (78.27%) were culture positive; in the rest 27(21.8%), diagnosis was based on Gram staining of the CSF. Streptococcus pneumoniae, Haemophilus influenzae, Neisseria meningitidis, Staphylococcus aureus and Escherichia coli were isolated in 33.9%, 33.9%, 5.6%, 2.4%, and 2.4% samples respectively. All the isolates had 100 percent; sensitivity to both ceftriaxone and ciprofloxacin while the sensitivities to penicillin and ampicillin were remarkably low. The mortality was 33/124 (26.6%) while 16/ 91 (17.6%) of the survivors had various neurologic sequelae. PMID- 16269840 TI - Tumor necrosis factor alpha and interleukin 6 in infants with sepsis. AB - Thirty-two infants above one month of age admitted to a tertiary care hospital with signs of infection and presumptive diagnosis of sepsis were included. Cytokine levels of tumor necrosis factor alpha (TNFmu) and interleukin-6 (IL 6) were estimated at admission and after 48 to 72 hr, and their relationship examined to the outcome. Significantly higher TNFalpha and IL-6 levels were seen in infants with sepsis compared to control. The TNFalpha levels significantly decreased in patients with sepsis, septic shock and the survivors, while the patients who did not survive, the levels showed no significant change after 48 hr. The initial levels of IL6 were comparatively higher in patients with septic shock and non-survivors, and increased at 48 hr of admission in patients with sepsis, septic shock and non-survivors. PMID- 16269842 TI - CSF Rhinorrhea and recurrent meningitis caused by transethmoidal meningoencephaloceles. AB - We report two children presenting with intermittent CSF rhinorrhea and recurrent meningitis. CT scan showed transethmoidal meningoencephalocele. PMID- 16269841 TI - Umbilical cord blood TSH levels in term neonates: a screening tool for congenital hypothyroidism. AB - This study was conducted to find normative values for thyroid stimulating hormone (TSH) in 1200 cord blood samples of term babies whose mothers were not on any thyroid medications. TSH was estimated within 24 hrs by enzyme immunoassay. A full thyroid profile, viz, T3, T4, TSH, fT3 and fT4 was done at 7-10 days of age in all babies with cord TSH >20 mIU/L. The mean, median and standard deviation for the TSH values for the cohort were 6.13 mIU/L, 5.8 mIU/L and 4.523 respectively. 22 babies with TSH values >20 mIU/L were given repeat tests. Hypothyroidism was confirmed in two of these babies. We conclude that a cut off value of TSH >20 mIU/L is adequate for neonatal thyroid screening in Indian settings. PMID- 16269843 TI - Congenital generalized lipodystrophy in a 4 year old Chinese girl. AB - Congenital generalized lipodystrophy (CGL) is a rare autosomal recessive disorder characterized by near complete absence of adipose tissue since birth and insulin resistance. The diagnosis is made on the basis of lack of body fat, muscular hypertrophy, acanthosis nigricans, hirsutism, hepatomegaly with fatty liver, hyperlipidemia and hyperglycemia with insulin resistance. We describe a 4-year old Chinese girl with the clinical features of CGL. PMID- 16269844 TI - Multiple sclerosis in childhood: presenting as bilateral optic neuritis. AB - Multiple sclerosis is a disease of adolescence and middle age. Though rare, it is being increasingly recognized in younger population. We describe two patients in whom multiple sclerosis was diagnosed before 10 years of age. Both patients had bilateral optic neuritis during the course of their illness with partial recovery; other lesions in the neuraxis recovered reasonably well. PMID- 16269845 TI - Kasabach-Merritt syndrome. PMID- 16269846 TI - Systematized nevus depigmentosus. PMID- 16269847 TI - Neonatal inguinal hernia. PMID- 16269848 TI - Acute flaccid paralysis: Guillain-Barre syndrome with enterovirus infection. PMID- 16269849 TI - Lactic acidosis in HIV infected children due to antiretroviral therapy. PMID- 16269850 TI - Onset of bilirubin rise in G6PD deficient neonates prenatal or postnatal. PMID- 16269851 TI - Chanarin Dorfman syndrome neonatal diagnosis and 3-year follow-up. PMID- 16269852 TI - Idiopathic pulmonary hemosiderosis. PMID- 16269854 TI - Hereditary methemoglobinemia in an infant. PMID- 16269853 TI - Idiopathic ileoileal intussusception in early infancy. PMID- 16269855 TI - Meconeum hydrocele presenting as a labial mass. PMID- 16269856 TI - What is appropriate post-exposure rabies prophylaxis? PMID- 16269858 TI - The current status of gene therapy in autologous transplantation. AB - Autologous hematopoietic cells have been used as targets of gene transfer, with applications in inherited disorders, cell therapy, and acquired immunodeficiency. The types of cells include hematopoietic progenitor cells, lymphocytes, and mesenchymal stem cells. The inherited disorders thus far approached in clinical trials include severe combined immunodeficiency, common variable gamma-chain immunodeficiency, chronic granulomatous disease, and Gaucher disease. Preclinical studies are vigorously under way in thalassemia, sickle cell anemia, Wiskott Aldrich syndrome and Fanconi anemia. Clinical trials of immunological therapy with gene-modified lymphocytes are under study in the treatment of malignancies. Clinical trials using anti-viral strategies for HIV infection in combination with autologous transplantation have begun, with additional approaches being developed. Gene therapy vectors are being developed to eliminate tumor cells contaminating autologous stem cell products. However, the risk of insertional mutagenesis and the potential for development of leukemia was highlighted by the first gene therapy trials in inherited immunodeficiency syndromes that achieved a therapeutic effect. Despite the slow progress of the field to date, there is extraordinary promise for gene therapy in the future. PMID- 16269860 TI - The significance of minimal residual disease in stem cell grafts and the role of purging: is it better to purge in vivo or in vitro? AB - Contamination of autologous graft by tumor, in addition to incomplete tumor eradication, can partly explain why relapse remains the commonest cause of treatment failure after autologous stem cell transplantation (ASCT) in patients with malignant hematologic disorders. Monitoring of minimal residual disease (MRD) is now recognized as an important diagnostic tool for assessment either of the response to treatments aimed at maximal cytoreduction and the individual risk of relapse. In order to improve cure rates, many strategies to achieve in vivo or in vitro reduction, if not eradication, of residual disease have been proposed. We discuss the significance of MRD and the role of purging in the ASCT setting, focusing on acute myeloid leukemia, chronic myeloid leukemia, multiple myeloma and follicular lymphoma. PMID- 16269859 TI - Role of CXCR4 chemokine receptor blockade using AMD3100 for mobilization of autologous hematopoietic progenitor cells. AB - G-CSF mobilization of hematopoietic progenitor cells (HPCs) is mediated through enzyme release from maturing myeloid cells, leading to digestion of adhesion molecules, trophic chemokines and their receptors, and the extracellular matrix. HPCs traffic to and are retained in the marrow through the trophic effects of the chemokine SDF-1alpha/CXCL12 binding to its receptor, CXCR4. AMD3100 reversibly inhibits SDF-1alpha/CXCR4 binding, and AMD3100 administration mobilizes CD34+ cells into the circulation. AMD3100 has been tested in several clinical trials which demonstrate that it improves the number of CD34+ cells mobilized including patients failing to mobilize with G-CSF alone. Engraftment of AMD3100-mobilized cells is prompt and durable. Toxicities are mild and infrequent. Lymphoma and myeloma cells do not appear to be mobilized. AMD3100 appears to be a promising agent for HPC mobilization. PMID- 16269861 TI - Allogeneic cell-mediated immunotherapy at the stage of minimal residual disease following high-dose chemotherapy supported by autologous stem cell transplantation. AB - Cumulative clinical experience suggests that immunotherapy may be an effective tool for eradicating tumor cells resistant to maximum tolerated doses of chemotherapy and radiation. Immunotherapy is much more effective when applied at the stage of minimal residual disease, especially against slowly growing tumors because development of graft-versus-leukemia, lymphoma, myeloma, or in a broader sense graft-versus-tumor effects renders immunotherapy more time consuming. Hence, eradication of rapidly growing bulky tumors may be difficult or impossible to achieve. Considering the fact that optimal immunotherapy may be accomplished in patients treated at the stage of minimal (MRD) disease, in patients with hematological malignancies and chemosensitive solid tumors a stage of MRD may be best achieved following administration of myeloablative high-dose chemotherapy or chemoradiotherapy supported by autologous stem cell transplantation (autoSCT). Taken together, immunotherapy following autoSCT may provide an ideal combination for improving the cure rate of otherwise incurable cancers, especially if tumor cells may respond to cytokine-mediated immunotherapy or cell-mediated cytokine activated immunotherapy. Following lymphocyte depletion in the course of autoSCT, adoptive transfer of alloreactive or tumor-reactive lymphocytes may be much more effective due to the preponderance of anticancer effector cells on the one hand, and elimination or depletion of the patient's regulatory cells that may downregulate anticancer effector mechanisms. PMID- 16269862 TI - Allogeneic hematopoietic cell transplantation as consolidation immunotherapy of cancer after autologous transplantation. AB - Autologous and allogeneic hematopoietic cell transplantations (HCT) after high dose conditioning are curative for many patients with hematologic malignancies. The major limitation of autologous HCT is disease recurrence, while the problem with allogeneic HCT is the restriction to relatively young and otherwise medically fit patients because of complications from high-dose therapy coupled with those from graft-versus-host disease (GVHD). The well-described benefits of graft-versus-tumor (GVT) effects have stimulated the development of conditioning regimens with attenuated doses of chemoradiotherapy that may provide both a degree of cytoreduction before allografting (reduced intensity) and facilitate allogeneic cell engraftment through immuno- suppression (nonmyeloablative). Both reduced-intensity and nonmyeloablative approaches seek to exploit GVT activity of donor cells to eradicate cancer. Because of intrinsic or acquired insensitivity of some malignancies to standard dose chemotherapy and the limitations of GVT effects in eradicating large tumor burdens, high-dose autologous HCT has been used before reduced-intensity and nonmyeloablative allogeneic HCT. This approach appears to improve response rates for malignancies such as multiple myeloma and lymphoma, while avoiding morbidity and mortality of high-dose allogeneic HCT. This review details the rationale, benefits and limitations of this approach. PMID- 16269863 TI - Improved conditioning regimens for autologous transplantation using targeted radiotherapy. AB - Most patients undergoing autologous hematopoietic stem cell transplantation for malignant diseases suffer recurrences of their neoplasms and die due to the inability of conventional high-dose conditioning regimens to eradicate their malignancies. As a result, intensive efforts to develop more effective conditioning regimens are currently under way at many institutions. Encouraging results have been obtained using targeted radiotherapy with radiolabeled antibodies or bone-seeking isotopes as components of novel conditioning regimens for autologous transplantation of patients with lymphomas, multiple myeloma and bone metastases. Results with radiolabeled antibodies targeting epithelial antigens on solid tumors, however, have been less encouraging. This report reviews the status of clinical studies using myeloablative doses of targeted radiotherapy in patients undergoing autologous stem cell transplantation for hematological malignancies or solid tumors. PMID- 16269864 TI - Autologous hematopoietic stem cell transplantation for autoimmune diseases. AB - Ten years have passed since the first published consensus statement on the use of hematopoietic stem cell transplantation (HSCT) in the treatment of severe autoimmune disease (AD) appeared. During that time, around 700 patients suffering from severe AD have undergone HSCT in the frame of phase I/II clinical trials from over 20 countries including the US. The majority have received an autologous HSCT using one of a limited number of regimens, consistent with the original consensus statement. Long-term drug-free remissions, remission then relapse, no response and treatment-related mortality (TRM) were seen in all the subgroups of AD. An overall TRM of 7% was observed, with marked variation between ADs, i.e. 11% in systemic lupus erythematosus (SLE) and only 1 patient in rheumatoid arthritis (RA). Phase III prospective, comparative randomized trials are running or being planned in multiple sclerosis (MS), systemic sclerosis (SSc), SLE and RA. Basic science programs are also being undertaken to study the immunological mechanisms underlying the clinical events observed. PMID- 16269865 TI - High-dose therapy for breast cancer - a case of suspended animation. AB - The role of high-dose chemotherapy in breast cancer is still controversial despite 20 years of clinical studies. Several nonrandomized studies had demonstrated improvement for patients with primary breast cancer. This led to the premature acceptance of high-dose therapy as a new standard of care for patients with high-risk breast cancer. There followed a phase of disillusionment after some of the randomized studies did not show any significant benefit and after a case of scientific misconduct. High-dose chemotherapy studies in breast cancer have been unpopular for the last 5 years. There is new evidence that warrants a new critical look. Fourteen randomized studies with a total of 5,592 patients have been carried out in patients with high-risk breast cancer on adjuvant therapy. Some of them showed significant improvement; others are coming to maturation now. In all randomized studies high-dose therapy in metastatic breast cancer leads to an equivalent or better disease-free survival, but because of their low power, none of these studies achieved an improvement in overall survival. It is thus necessary to perform a meta-analysis of all these studies to acquire insight into the choice of high-dose regimens. It is further necessary to look at the biology of breast cancer in the context of high-dose chemotherapy studies. PMID- 16269866 TI - Autologous hematopoietic stem cell transplantation in Non-Hodgkin's lymphomas. AB - Early studies on high-dose therapy followed by autologous bone marrow transplantation demonstrated the safety and efficacy of this approach in non Hodgkin's lymphoma (NHL). As the procedure became easier and safer with the introduction of peripheral blood progenitor cells instead of marrow, the popularity of this procedure declined. In the last decade, autologous hematopoietic stem cell transplantation has played an important role in front line and salvage treatments of indolent and aggressive NHL. PMID- 16269867 TI - Phenomenological evidence for two types of paranoia. AB - OBJECTIVE: Two types of paranoia have been identified, namely persecution (or 'Poor Me') paranoia, and punishment (or 'Bad Me') paranoia. This research tests predicted differences in phenomenology--specifically, in person evaluative beliefs, self-esteem, depression, anxiety, and anger. METHOD: Fifty-three people with current paranoid beliefs were classified as Poor Me, Bad Me, or neither (classification was reliable). Key dependent variables were measured. RESULTS: All predictions were supported, except the one relating to anger, where the two groups did not differ. The Bad Me group had lower self-esteem, more negative self evaluative thinking, lower negative evaluations about others, higher depression and anxiety. Importantly, the differences in self-esteem and self-evaluations were not fully accounted for by differences in depression. CONCLUSION: Data support the presence of two distinct topographies of paranoia. Future research is needed to explore the theory further and examine clinical implications. PMID- 16269868 TI - Charles Dickens' A Tale of Two Cities: a case report of posttraumatic stress disorder. AB - In posttraumatic stress disorder, a traumatic event is persistently re experienced in the form of intrusive recollections, dreams or dissociative flashback episodes; cues to the event lead to distress and are avoided, and there are persistent symptoms of increased arousal. While this diagnostic concept has been widely discussed and its existence questioned, a novel written by Charles Dickens long before it was included in any diagnostic system can be viewed as an early case report of posttraumatic stress disorder. PMID- 16269869 TI - Delusions in individuals with schizophrenia: factor structure, clinical correlates, and putative neurobiology. AB - BACKGROUND: Delusions are a central feature of schizophrenia, yet our understanding of their neurobiology is limited. Attempt to link dimensions of psychopathology to putative neurobiological mechanisms depends on careful delineation of symptoms. Previous factor analytic studies of delusions in schizophrenia were limited by several methodological problems, including the use of patients medicated with antipsychotics, inclusion of nondelusion symptoms in the analyses, and/or inclusion of patients with psychotic disorders other than schizophrenia. These problems may have possibly biased the resulting factor structure and contributed to the inconclusive findings regarding the neurobiology of positive symptoms. Our goal is to examine the factor structure of delusions in antipsychotic-free individuals with diagnoses of schizophrenia/schizoaffective disorder. SAMPLING AND METHODS: We assessed 83 antipsychotic-free individuals with DSM-IV diagnoses of schizophrenia/schizoaffective disorder. A principal component analysis was conducted on the delusions symptoms of the SAPS. RESULTS: The principal component analysis resulted in three distinct and interpretable factors explaining 58.3% of the variance. The Delusions of Influence factor was comprised by delusions of being controlled, thought withdrawal, thought broadcasting, thought insertion, and mind reading. The Self-Significance Delusions factor was comprised by delusions of grandeur, reference, religious, and delusions of guilt/sin. The Delusions of Persecution factor was comprised solely by persecutory delusions. The three factors displayed distinct associations with hallucinations, bizarre behavior, attention, positive formal thought disorder, and avolition/apathy. CONCLUSIONS: The results indicate that delusions are best described by three distinct subtypes. The authors propose a novel model linking the three delusion subtypes, attributions to self/other, and putative neurobiological mechanisms. Implications for future research are discussed, as well as links to cognitive-behavioral conceptualizations of delusions. PMID- 16269870 TI - Phytoestrogen resveratrol suppresses steroidogenesis by rat adrenocortical cells by inhibiting cytochrome P450 c21-hydroxylase. AB - BACKGROUND AND AIM: The phytoestrogen resveratrol is found in grapes, mulberries and peanuts, all of which are consumed regularly by humans. Resveratrol is also used in chemotherapy against cancer and aging and as a cardioprotectant. The aim of the present study was to characterize the effects of resveratrol on rat adrenal steroidogenesis and to study the underlying mechanism. METHODS: Adrenocortical cells were isolated from the adrenal glands of normal male rats (in vitro) and from male rats administered resveratrol in their diet for 12 weeks (ex vivo). Cells from resveratrol-treated and non-treated rats were tested ex vivo for responsiveness to ACTH and cells from normal rats were tested in vitro for responsiveness to ACTH in the presence and absence of resveratrol. Corticosterone and progesterone production were measured by RIA and expression of steroidogenic enzymes analyzed by PAGE/Western blotting. RESULTS: Corticosterone production was inhibited 47% by 50 microM resveratrol in vitro and 20% ex vivo, while progesterone production was elevated to 400% of the control value in in vitro experiments. Resveratrol treatment decreased adrenal cytochrome P450 c21 hydroxylase expression in vivo and cell culture conditions. No changes in cell viability or morphology were caused by exposure to resveratrol in both ex vivo and in vitro experiments. CONCLUSION: Resveratrol suppresses corticosterone production by primary rat adrenocortical cell cultures in vitro and ex vivo by inhibiting cytochrome P450 c21-hydroxylase. PMID- 16269871 TI - Rate of recalls in congenital hypothyroidism based upon a regional survey in Isfahan, Iran, using serum T4 and TSH analyses: comparison of two different recall methods. AB - AIMS: To evaluate and compare the recall rate in congenital hypothyroidism screening project in Isfahan, first using an approach involving measures of both TSH and T4 and then using TSH alone. METHODS: From June 2002 to January 2005, serum TSH and T4 level of referred neonates were measured at 3rd to 7th day of birth through venous sampling. If neonates' serum TSH was >20 mIU/l or T4 was <6.5 microg/dl by the first protocol, or TSH was >20 mIU/l by the second protocol, they were recalled. TSH and T4 were measured using an immunoradiometric assay and radioimmunoassay, respectively. Neonates with TSH > 10 and T4 < 6.5 on their second measurement were considered as congenitally hypothyroid. RESULTS: Serum T4 and TSH of 29,425 neonates by first and 57,235 neonates by second recall approach were measured. Recall rate was higher in the first protocol (2.2% vs. 0.6%, p < 0.05). Most of the recalled neonates in the first protocol were recalled for low T4 level (p < 0.05). The prevalence of CH was 1 in 350 livebirths. CONCLUSION: Although the recall rate was in the acceptable range by either approach, the TSH alone protocol seems to be a more sensitive and practical approach with the least recall burden and considering the high prevalence of CH in our region merit adaptation of widespread screening for CH using TSH measurements from heel stab blood spotted on filter paper. PMID- 16269872 TI - Effects on bone mineral density by treatment of benign nodular goiter with mildly suppressive doses of L-thyroxine in a cohort women study. AB - OBJECTIVES: Thyroid diseases and their treatment may influence the osseous system. The influence that prolonged suppressive L-thyroxine (LT4) therapy may have on inducing subclinical hyperthyroidism on bone metabolism is still a matter of debate. The aim of the present study was to assess the effects of chronic LT4 treatment at mildly inhibiting serum thyroid-stimulating hormone (TSH) doses on bone mineral density (BMD) and biochemical bone remodeling markers in a cohort of women with benign nodular goiter, and to verify the efficacy of the treatment on nodule size. SUBJECTS AND STUDY DESIGN: A total of 200 euthyroid Caucasian women with nodular goiter (age 52.1 +/- 9; 80 pre- and 120 postmenopausal) were enrolled: 96 had been treated with LT4 for at least 3 years and a matched group of 104 had untreated goiter. LT4 therapy was given at a dose sufficient to reduce TSH under the lower limit of the normal range (0.27-4.20 microIU/ml) without suppressing it below the limit of assay sensitivity (0.005 microIU/ml) and maintaining normal serum values of free triiodothyronine (FT3) and free thyroxine (FT4). The adequacy of the dose was evaluated on the basis of serum TSH levels. The osteopenic effect of LT4 treatment was evaluated directly by total body and lumbar spine dual-energy X-ray absorptiometry (DEXA) and indirectly by biochemical parameters (alkaline phosphatase, osteocalcin, calcium, parathyroid hormone) at the baseline and throughout the follow-up. The efficacy of LT4 schedule on thyroid nodule size was assessed on the basis of the ultrasonographic evaluation. RESULTS: Mineralometric data showed no significant difference between BMD values for treated and untreated patients in both pre- and postmenopausal status. In all patients, serum markers of bone turnover were in the normal range, with no differences in the treated and control groups. The TSH concentrations were significantly lower in treated than in untreated patients (p < 0.0001); FT3 and FT4 were in the normal range for all patients. Evaluation of nodule size during follow-up showed a reduction of > or = 30% in 32 of 96 treated patients (33.3%) versus none in those untreated, whilst nodule size remained unmodified in 60 treated patients (62.5%) versus 35 (33.6%) in those untreated, and an increase in nodule size and/or development of new nodules was found in 4 treated patients (4.2%) versus 69 of the 104 untreated patients (66.3%). CONCLUSIONS: This study suggests that at slightly suppressing TSH doses, LT4 therapy has no adverse effects on BMD in both pre- and postmenopausal women, while having an efficacy on nodule size comparable with that reported using an LT4 schedule able to maintain TSH near or below the assay sensitivity limit. PMID- 16269873 TI - Pegvisomant treatment in a 4-year-old girl with neurofibromatosis type 1. AB - BACKGROUND/AIMS: Growth hormone (GH) excess in childhood is a rare disorder. Current treatment options such as somatostatin analogues, pituitary surgery or irradiation can have serious side effects. Recently, a GH receptor antagonist, pegvisomant, was introduced for the treatment of adults with acromegaly. We wanted to investigate whether pegvisomant was effective in a child with octreotide-resistant GH excess. CASE: A 4-year-old girl with neurofibromatosis type 1 and GH excess associated with optic glioma received pegvisomant injections (10 mg subcutaneously) with increasing intervals from daily to every 4th day. RESULTS: IGF-I and IGFBP-3 decreased from +6.9 and 4.6 standard deviation scores (SDS), respectively, to within normal range. Height velocity dropped from 12.4 SDS to mean -0.7 SDS (range: -5.0 to 5.0) and height SDS decreased from +1.3 to +0.6 (target height: +0.2). Random non-fasting serum GH values were mean 5.0 mlU/l (range: 1.6-9.5). There was no change in fasting blood glucose (4.6-4.7 mmol/l) or glycosylated haemoglobin (5.5%) and no subjective or biochemical side effects. Repeated tests of thyroid, adrenal and gonadal function showed no alterations during the treatment period. Intracranial tumours remained unchanged in size and visual impairment did not deteriorate. CONCLUSION: Pegvisomant normalized IGF-I and IGFBP-3 levels. Growth velocity was normalized after initial catch-down growth, and it remains to be seen whether this result can be maintained during long-term treatment. PMID- 16269874 TI - [Treatment of intertrochanteric femur fractures in elderly patients: internal fixation or hemiarthroplasty]. AB - OBJECTIVES: There is no consensus as to whether internal fixation or hemiarthroplasty is more appropriate for the treatment of intertrochanteric femur fractures in elderly patients. While the latter offers early mobilization, internal fixation preserves the hip joint and avoids long-term complications associated with the prosthesis. This retrospective study aimed to compare the early results of these treatment modalities. METHODS: The study included 81 patients who were available for follow-up after surgery for intertrochanteric femur fractures. Of 38 patients (mean age 77.7 years; range 65 to 99 years) treated with internal fixation, 25 were alive; of 43 patients (mean age 80 years; range 67 to 97 years) treated with hemiarthroplasty, 22 were alive at the last follow-ups. The two groups were compared with regard to perioperative characteristics, mobilization time, complications, mortality, and daily activities according to the Barthel Activities of Daily Living Index. The mean follow-up was 22.7 months (range 6 to 39 months) in internal fixation, and 22.3 months (range 7 to 39 months) in hemiarthroplasty groups. RESULTS: Subsequent to the operation, mortality occurred in 34.2% after a mean of 13 months (range 1 to 36 months) and in 48.8% after a mean of six months (range 1 to 24 months) in patients treated with internal fixation and endoprosthesis, respectively. There were no significant differences with respect to mobilization in bed, standing, weight bearing without support, complications, and daily activity scores. The only significant difference in favor of hemiarthroplasty was that full weight bearing with two crutches took a shorter time (p<0.05). CONCLUSION: Short-term results suggest that hemiarthroplasty is not an advantageous alternative to internal fixation; moreover, its postoperative survival is shorter and mortality rate is higher. Osteosynthesis seems to be the first choice in the treatment of elderly patients with intertrochanteric femur fractures. PMID- 16269875 TI - [Risk factors for osteoporosis in women having hip fractures after 60 years of age]. AB - OBJECTIVES: Management of hip fractures in elderly women should not be confined to surgical treatment and rehabilitation, but also encompasses interventions and measures to protect them from hip fractures. We administered a questionnaire to female patients with hip fractures to determine their social profiles and risk factors. METHODS: A total of 107 female patients (mean age 74 years; range 63 to 100 years) who experienced hip fractures after the age of 60 years were administered a questionnaire to determine their physical and social characteristics, medical conditions , and nutritional status. In addition, the types of fractures and the Singh index were determined on hip radiographs. Femoral neck (n=51, 47.7%) and intertrochanteric (n=56, 52.3%) fractures were classified according to the Garden and modified Evans classifications, respectively. RESULTS: The mean body mass index was 23 kg/m2. The Singh index was grade 3 in 70.1%, grade 2 in 26.2%, grade 4 in 2.8%, and grade 1 in 0.9%. Forty four patients (41.1%) were illiterate, and 31 patients (29%) were only literate or could finish primary school. A history of smoking was found in 29.9% for a mean duration of 30.7 years. The majority of patients (67.3%) had limited activity and 63.6% had no outdoor work at all. None of them had a regular sport activity. Only two patients (1.9%) received inadequate and short-term postmenopausal therapy. The mean daily milk consumption was less than a glass (125-150 ml). Calcium supplements were used in only 5.6%. A history of previous fractures (hip, vertebra, distal radius) was detected in 9.3%. The most common coexistent diseases were hypertension (50%) and diabetes (29%). None of the patients had bone mineral density measurements. CONCLUSION: Our study showed that, despite the high incidence of risk factors for osteoporosis, few patients received medical care for the diagnosis and treatment of osteoporosis, and that preventive health care measures were not available for this patient group. PMID- 16269876 TI - [Nonvascularized fibular grafts in the reconstruction of bone defects in orthopedic oncology]. AB - OBJECTIVES: Bone defects after resection or curettage of musculoskeletal tumors are commonly reconstructed with the use of structural fibular autografts or allografts with considerable success. In this study, we compared the clinical and radiographic results of nonvascularized autografts and allografts. METHODS: Fifty seven patients (30 males, 27 females; mean age 22.8 years; range 5 to 57 years) underwent reconstruction with a structural fibular autograft (n=30) or allograft (n=27) following tumor resection. Of the tumor defects, 24 were segmental and 33 were cavitary. All the patients who had segmental reconstruction were treated with internal fixation, except for one patient who had internal fixation and monolateral external fixator for femoral reconstruction. The results were evaluated with respect to union, time to union, and complications. The mean follow-up period was 46 months; 48.4 and 42.3 months in the allograft and autograft groups, respectively. RESULTS: Radiographically, union was obtained in 46 patients (80.7%) within a mean of 5.9 months (6.8 months in 20 autografts, and 5.1 months in 26 allografts). Nonunion (19.3%) occurred in four allografts and seven autografts. No significant difference was found between the two fibular grafts in terms of union (p>0.05). Graft rupture was detected in six patients (5 autografts, 1 allograft) all of which were due to minor traumas. Complications were encountered in eight patients (14%) including transient peroneal nerve injury (n=3), wrist subluxation (n=2), infection (n=2), and Madelung's deformity. CONCLUSION: Reconstruction of cavitary and segmental bone defects with autogenous or allogenous nonvascularized fibular grafts is a reliable method, with high success rates. PMID- 16269877 TI - [Diagnosis of septic arthritis and initial antibiotic treatment]. AB - OBJECTIVES: This study was designed to determine the similarities and differences in clinical, laboratory and radiographic presentation of septic arthritis in childhood and at adult ages, to find out its etiological profile, and to establish an antibiotic treatment protocol for the initial period and for patients in whom the causative agent could not be identified. METHODS: Thirty four patients (age range 15 months to 85 years) who underwent surgery with a diagnosis of septic arthritis were retrospectively studied in two groups, namely, children-adolescents (age = or < 15 years ; 16 patients) and adults (age >15 years; 18 patients). Clinical and laboratory findings of septic arthritis were compared with operation findings. The etiological profile and sensitivity patterns were investigated. RESULTS: Unflatering features in both groups were clinical findings of decreased range of motion and tenderness, laboratory findings of elevated erythrocyte sedimentation rate and C-reactive protein, and domination of polymorphonuclear leukocytes in the joint fluid. Gram staining of the joint fluid was highly informative in terms of probable bacteria. During the first two years of life, the most common bacteria were H. influenzae and S. pneumoniae, and after two years, staphylococci and streptococci. Ciprofloxacin and sulbactam-ampicillin were found effective against most of the Gram-positive microorganisms isolated in both groups. CONCLUSION: The most useful test for septic arthritis is arthrosynthesis and macroscopic and microscopic analyses of the material. Gram staining is of great help in the planning of initial antibiotic treatment. For patients older than two years of age, treatment with sulbactam-ampicillin is effective against staphylococci and streptococci, and amikacin against Gram-negative bacteria. Detection of Gram-negative bacteria in patients younger than two years should bring H. influenzae in mind, for which ampicillin must be the first choice. PMID- 16269878 TI - [Talectomy for the treatment of neglected pes equinovarus deformity in patients with neuromuscular involvement]. AB - OBJECTIVES: We evaluated talectomy operations performed for the correction of neglected pes equinovarus deformity of the foot in patients with neuromuscular involvement. METHODS: Eleven patients (2 females, 9 males; mean age 9.5 years; range 3 to 20 years) with various neurological disorders underwent 17 talectomy operations for the correction of neglected pes equinovarus deformity. Diagnoses were spina bifida in seven patients, arthrogryposis multiplex congenita in two patients, and spinal muscular atrophy and poliomyelitis in two patients, respectively. None of the patients had a previous surgery for pes equinovarus deformity. Preoperatively, three patients and one patient could walk in home settings with and without support, respectively, whereas seven patients could not walk at all. A protective orthosis was used in all the patients postoperatively. Achillotomy and posterior capsulotomy were performed prior to talectomy when needed. The results were evaluated with the use of a functional rating system and radiographs. The mean follow-up was 50 months (range 24 to 108 months). RESULTS: The results were good in 12 feet and fair in five feet. Seven patients could walk without support, and three patients could walk with support. One patient required re-operation for residual deformity of the foot and skin wounds due to poor compliance with the protective orthosis. One patient could not ambulate despite a plantigrade foot. CONCLUSION: Talectomy is a limb-saving procedure for the treatment of neglected pes equinovarus deformity. Its success relies not only on obtaining a plantigrade foot but also on a proper postoperative orthotic control. PMID- 16269879 TI - [The effect of injury level, associated injuries, the type of nerve repair, and age on the prognosis of patients with median and ulnar nerve injuries]. AB - OBJECTIVES: In this study, we aimed to evaluate the functional results of nerve repair (median and/or ulnar) in patients with forearm clean-cut injuries and investigated the effect of injury level, associated injuries, the type of repair (primary or secondary), and age on the prognosis. METHODS: The study included 42 patients (34 males, 8 females; mean age 31 years; range 9 to 62 years) who were treated for forearm clean-cut injuries. Involvement was in the proximal forearm in four, mid-forearm in 11, and distal forearm in 27 cases. There were 51 nerve injuries affecting the median nerve (n=30) and the ulnar nerve (n=21). Nerve injuries were isolated in 12 patients, associated with tendon injuries in nine patients, and with tendon and artery injuries in 21 patients. The patients were evaluated in four age groups including 0-15, 16-30, 31-45 years, and 46 years or above. Functional evaluations were made using the Seddon classification. The effect of injury level, associated injuries, the type of repair, and age on the prognosis was assessed. The mean follow-up was 39 months (range 11 to 57 months). RESULTS: Although the clinical and functional results of primary and late-primary repairs were less favorable than those of secondary repairs, the difference did not reach a significant level (p>0.05). The injury level, associated injuries, and age did not influence the Seddon scores significantly (p>0.05). In the age group of 0-15 years, the results were very good in all the patients (100%), but good and very good results accounted for only 20% in the age group of 46 years or above. CONCLUSION: In appropriate cases with clean-cut nerve injuries, primary repair must be the first choice. Taking the low regeneration capacity into consideration, priority should be given to reconstructive procedures in patients at older ages. PMID- 16269880 TI - [External rotation osteotomy of the humerus for the treatment of shoulder problems secondary to obstetric brachial plexus palsy]. AB - OBJECTIVES: We evaluated the results of external rotation osteotomy of the humerus for the treatment of shoulder problems secondary to obstetric brachial plexus palsy. METHODS: Forty children (24 boys, 16 girls; mean age 7.5 years; range 23 months to 14.8 years) underwent external rotation osteotomy of the humerus. Involvement was at the C5-6 roots in 11 patients, C5-6-7 roots in 19 patients, and at all the roots in 10 patients. Twenty-six patients had humeral head deformity and eight patients had posterior subluxation. The shoulder was ankylosed in one patient. Zancolli and Putti signs were positive in six and eight patients, respectively. The mean active shoulder abduction was 80 degrees (range 0 to 170 degrees ) and the mean internal rotation contracture was 27 degrees (range 10 degrees to 50 degrees ). Fourteen patients, all of whom were beyond five years of age, had an abduction contracture. Preoperative and postoperative functional evaluations were made with the use of the Mallet scale. Preoperatively, 35 shoulders had a score of II, five had a score of III. Osteotomies were performed in the proximal humerus in patients older than five years and in the mid-humerus in those without a contracture or younger than five years. RESULTS: The mean postoperative shoulder abduction was 95.7 degrees (range 30 degrees to 170 degrees ). Internal rotation contractures improved in all the patients. Abduction contractures did not resolve in two patients in whom a mid humeral osteotomy was performed. Postoperative Mallet scores ranged from II to V in five, six, 15, and 14 shoulders, respectively. Rotation provided by the osteotomy was lost in one patient because of a humerus fracture that occurred in a traffic accident. Passive total shoulder rotation remained unchanged following surgery. Patients having better preoperative range of motion and who were at younger ages benefited the most from surgical treatment. CONCLUSION: External rotation osteotomy of the humerus must be performed at early ages before the shoulder gets stiffer. PMID- 16269881 TI - [Treatment of medial epicondyle fractures accompanying elbow dislocations in children]. AB - OBJECTIVES: We evaluated the clinical and radiographic results of treatment for medial epicondyle fractures accompanying elbow dislocations in children. METHODS: The study included 10 children (9 boys, 1 girl; mean age 12 years; range 1.5 to 15 years) with medial epicondyle fractures accompanying elbow dislocations. Three patients had posteromedial and seven patients had posterolateral dislocations. Three patients were treated conservatively, while four patients and three patients underwent early and late surgical treatment before or after the seventh day of injury, respectively. Surgery was indicated due to entrapment of the medial epicondyle fragments in three patients, and to instability in four patients with more than 5 mm of displacement. At surgery, a posteromedial incision was used and ulnar nerve exploration was performed. The results were evaluated using the Mayo elbow performance score. The mean follow-up period was 28 months (range 3 to 103 months). RESULTS: Union of the medial epicondyle fractures was achieved between four to six weeks in all the patients. The mean Mayo elbow performance score was 93.5. Late surgery was associated with a score of 80 in two patients and 75 in one patient, the remaining patients had an excellent result (100 points). Full range of elbow motion was achieved in all the patients treated conservatively and with early surgery; however, following late surgery, two patients had extension and flexion losses of 5 degrees and 10 degrees , respectively. None of the patients had instability postoperatively. CONCLUSION: Patients with entrapment of the medial epicondylar fragment in the joint and with a displacement of more than 5 mm should undergo surgery, while those with a displacement of 5 mm or less can be treated conservatively. PMID- 16269882 TI - [The effect of neostigmine and atropine combination on postoperative nausea and vomiting after arthroscopic surgery]. AB - OBJECTIVES: Postoperative nausea and vomiting prolong the time spent in postanesthesia recovery units. In this study, we investigated the effect of neostigmine and atropine combination, used to avoid residual curarization, on nausea and vomiting. METHODS: The study included 40 ASA I-II patients who were planned to have a short-term arthroscopic operation. The patients were administered a single dose of 0.4 mg/kg atracurium besylate for muscle relaxation, and then, were randomly divided into two groups. In group I, neuromuscular blockade was eliminated with 1.5 mg neostigmine and 0.5 mg atropine, whereas group II patients underwent spontaneous resolution. The patients were evaluated for nausea and vomiting and the need for antiemetic drugs in the recovery unit, patient room, and on the postoperative second day. RESULTS: There were no significant differences between the two groups with respect to hemodynamic parameters and peripheral oxygen saturation. The mean operation time did not differ significantly (p>0.05), but the mean extubation time was significantly shorter in group I (p<0.05). No significant differences were observed for the occurrence of nausea and vomiting and the need for antiemetic drugs in the recovery unit, patient room, and on the postoperative second day (p>0.05). CONCLUSION: In patients undergoing arthroscopic surgery, it is safe to use neostigmine and atropine combination before extubation to avoid residual neuromuscular blockade associated with the use of non-depolarizing myorelaxants. PMID- 16269883 TI - [A survey on knowledge, attitude, and practice of emergency department practitioners in musculoskeletal injuries]. AB - OBJECTIVES: A knowledge, attitude, and practice study was conducted in relation to musculoskeletal injuries among emergency department practitioners working in the town of Afyon, Turkey. METHODS: Of 40 practitioners working at emergency health services in Afyon, 32 (80%) responded to a 20-item questionnaire that inquired about their knowledge, attitude and practice. A 100-point scoring system was used for the first half of the questionnaire including 10 questions about their knowledge, and a total score of 60 or above was regarded as successful. Inquiry was focused on frequent emergency problems that require intervention by the emergency department practitioners, such as neck examination in trauma patients, treatment of open fractures, intervention to dislocations, treatment in gunshot injuries, and transportation of traumatic amputated organs. RESULTS: The mean duration of professional practice was 7.8+/-3.9 years. The mean achievement score was 66.6+/-15.8, without any significant difference among institutions. No significant relationship existed between the duration of professional practice and knowledge scores. Despite a success rate of 81.3% in knowledge scores, the following shortcomings were notable: neglect in the neurological examination of the lower extremity during neck examination, and inappropriate definitions and interventions in open fractures, tetanus prophylaxis, and antibiotic treatment. CONCLUSION: The survey showed that the practitioners in Afyon have sufficient knowledge levels; however, it seems apparent that there are shortcomings in their practice, which require continuing education for all the staff. PMID- 16269884 TI - [Biomechanical analysis of load transmission characteristics of limited carpal fusions used to treat Kienbock's disease]. AB - OBJECTIVES: Although limited carpal fusions used in the treatment of Kienbock's disease are thought to act by decreasing the loads on the lunate, biomechanical studies show that capitohamate fusion acts oppositely to what is expected. This experimental study was designed to resolve this paradox. METHODS: In a biomechanical cadaveric study, load transmissions at the radioulnacarpal joint were investigated under 140 and 210 newtons of load with three wrist postures, namely, neutral, ulnar and radial deviations, in five intact wrists and after scaphotrapeziotrapezoid, capitohamate, and scaphocapitate fusions. RESULTS: Under 140 newtons of load, the loads imposed to the lunate decreased following scaphotrapeziotrapezoid and scaphocapitate fusions, but increased after capitohamate fusion. However, when the load was increased to 210 newtons, there were no differences between intact wrists and limited carpal fusions in respect to the loads exerted on the lunate. In all the situations, the lunate was subjected to a significantly greater load in ulnar deviation. CONCLUSION: These results suggest that limited carpal fusions do not alter load transmission characteristics of the wrist joint under 210 newtons of load. The etiology of the Kienbock's disease seems to be related to an overload in ulnar deviation and the beneficial effect of limited carpal fusions seems to be associated with restricted ulnar deviation of the wrist rather than load transmission characteristics. PMID- 16269885 TI - [The effect of the tibial tunnel fixation level on the results of cyclic loading in anterior cruciate ligament reconstruction]. AB - OBJECTIVES: This study was designed to compare the biomechanical characteristics of non-anatomic (far from joint) and anatomic (close to joint) levels of tibial tunnel fixation with soft tissue graft using a soft tissue interference screw in anterior cruciate ligament (ACL) reconstruction. METHODS: Twelve bovine tibiae and digital extensor tendons were divided into two homogeneously equal groups after removing soft tissues. Tibial tunnels were prepared with a 7-mm drill with the use of an ACL guide adjusted to 45 degrees . Each tunnel was then dilated to 9 mm in 0.5 mm increments. Digital extensor tendons were fixed at non-anatomic (group I) or anatomic (group II) tibial tunnel levels with a soft tissue metal interference screw, 9 x 30 mm in size. All the specimens were cycled 500 times from 50 to 250 N with 1 Hz frequency in a servo-hydraulic testing machine followed by ultimate load at-failure testing at a rate of 20 mm/min. Statistical analyses were made using the Mann-Whitney U-test. RESULTS: The mean screw insertion torque values were 8.2+/-2.4 Nm and 7.8+/-2.3 Nm in groups I and II, respectively (p=0.88). The mean values of graft displacement (1.9+/-0.75 mm versus 2.2+/-1.2 mm, p=0.63) and stiffness (132.72+/-10.93 N/mm versus 125.14+/ 15.93 N/mm, p=0.63) did not differ significantly. CONCLUSION: The biomechanical properties of ACL reconstruction with soft tissue graft fixation using a soft tissue interference screw are not influenced by the level of tibial tunnel fixation. PMID- 16269886 TI - [Phalanx-like osseous structure posterior to the cervical vertebrae: a hitherto unreported location for a rare anomaly]. AB - Several types of abnormalities of the vertebral column have been reported. Pelvic rib or pelvic digit is a rare congenital anomaly of the spine. A 58-year-old male patient presented with a complaint of a firm mass in the neck. He had no history of trauma and had been aware of the mass for years. Physical examination showed a painless and immobile mass in the neck, measuring 4 x 1 cm. About 10 to 15 degrees of restriction was noted in neck movements, with slight pain. A plain radiograph of the cervical region showed two bony formations resembling a coccygeal segment or a phalanx, posterior to the spinous process of the C6 vertebra. They had well-defined cortices and medullae and pseudo-articulation between each other and with the spinous process. These findings were confirmed by three-dimensional computed tomography and magnetic resonance images. The patient did not accept surgery for the lesions were not associated with a significant discomfort. To our knowledge, such a digit has hitherto not been reported in the cervical region. PMID- 16269887 TI - [Familial glomus tumor encountered in the same finger and localization in four family members]. AB - A 35-year-old man was referred to our clinic by a neurosurgeon for hypersensitivity and severe pain in his right middle finger. History showed that the symptoms had been present for three years and he had made several attempts to seek medical attention. A sensitive, very painful mass was palpated under the eponychium of his right middle finger. The tumor was surgically removed and histological examination confirmed the clinical diagnosis of a glomus tumor. Inquiry into the family history revealed that the same problem existed in the same localization with similar complaints in the father and two daughters. Both the records of the father's operation 10 years before and clinical-radiological findings of the daughters were consistent with a familial entity. In the presence of hypersensitivity and pain in the finger, family members should be inquired and a diagnosis of familial glomus tumor should be considered. PMID- 16269888 TI - [Van Nes rotationplasty in two patients with congenital femoral deficiency]. AB - Congenital femoral deficiencies have a wide spectrum ranging from simple hypoplasia to complete femoral aplasia. They are often associated with congenital shortening of the tibia and fibular hemimelia. This anomaly has no known genetic transmission. There are two main treatment modalities for congenital femoral deficiency: prosthetic replacement surgery (Syme amputation or Van Nes rotationplasty followed by prosthetic fitting) and lengthening reconstruction surgery. In this report, we presented two patients (male, 26 years; girl, 7 years) with congenital femoral deficiency treated with Van Nes rotationplasty. In both cases, the treatment took 1.5 months, after which the patients were mobilized with prosthesis. No complications were encountered within a follow-up period of five years and 1.5 years, respectively. Rotationplasty enables an improved functional gait in patients with a very short femoral segment or Paley type 3 femoral deficiency. PMID- 16269890 TI - Acute stroke: pathophysiology, diagnosis, and treatment. AB - Stroke, a neurologic event due to altered cerebral circulation, is the third leading cause of death in the United States. Risk factors for stroke include hypertension, family history, and diabetes mellitus. The subtypes of stroke are ischemia, infarction, and hemorrhage. Ischemia and infarction are the result of atherosclerotic development of thrombi and emboli. Decreased and/or absent cerebral circulation causes neuronal cellular injury and death. Intracerebral hemorrhage occurs from rupture of cerebral vessels often as the result of hypertension. Patient assessment and diagnosis include the use of computed tomography scans, magnetic resonance imaging, and the National Institute of Health Stroke Scale, and treatment depends on the etiology of the stroke. Thrombolytic therapy is the mainstay of treatment for thrombotic and embolic events. Current recommendations for future stroke care include the development of designated stroke centers. Directions for research in stroke treatment includes examining neuroprotective therapies. PMID- 16269892 TI - Intracranial pressure monitoring: why monitor? AB - Evidence suggests that the mortality and morbidity of acquired brain injury could be reduced if clinicians used an aggressive intracranial pressure guided approach to care. Despite nearly 50 years of evidence that intracranial pressure monitoring benefits patient care, only about half of the patients who could benefit are monitored. Some clinicians express concerns regarding risks such as bleeding, infections, and inaccuracy of the technology. Others cite cost as the reason. This article discusses the risks and benefits of intracranial pressure monitoring and the current state of evidence of why patients should be monitored. PMID- 16269891 TI - Consciousness, coma, and caring for the brain-injured patient. AB - In this article, a preliminary conceptual framework is presented for exploring nursing interventions and research aimed at improving care of the unconscious brain-injured patient during the early subacute phase of brain injury. The cue response framework presented is derived from multidisciplinary sources and has specific clinical relevance to critical care nurses caring for unconscious brain injured patients. A key aspect of this framework is the attention focused on the timing of nursing interventions in response to how nurses interpret the physical, physiological, and secondary cues they observe when caring for comatose patients. A case exemplar is used to present one example of how this framework may be used in the clinical setting. PMID- 16269893 TI - Cerebral blood flow monitoring in clinical practice. AB - The brain depends on a continuous flow of blood to provide it with oxygen and glucose needed to maintain normal function and structural integrity, thus cerebral blood flow is normally tightly regulated. A decrease in cerebral blood flow to ischemic levels may be tolerated for only minutes to hours, depending on the severity of the ischemia. If cerebral blood flow ceases completely, brain cell death occurs within minutes. A variety of conditions are encountered clinically, such as stroke or traumatic brain injury, where an actual or potential alteration in cerebral blood flow puts the brain at risk for ischemia and infarction. In this article, the physiology of cerebral blood flow will be presented as a basis for understanding cerebral blood flow regulation and the rationale for clinical interventions to optimize cerebral blood flow. Techniques currently available to assess cerebral blood flow and clinical situations in which cerebral blood flow is measured will be discussed. Clinical interventions will be presented briefly. PMID- 16269894 TI - The effect of hypothermia and hyperthermia on acute brain injury. AB - The brain is extraordinarily susceptible to changes in temperature. Hyperthermia has been shown to exacerbate the biochemical cascade of secondary brain injury. Inversely, hypothermia limits the damaging effects of secondary brain injury. There has been a great deal of investigation regarding the detrimental effects of hyperthermia and the neuroprotection of hypothermia in animal studies. Within the last decade, clinical trials have begun to establish how the brain reacts to both temperature extremes. In the future, studies of hypothermia will continue in the quest of the optimal timing and degree of hypothermia. Hyperthermia will be examined in depth for its detrimental effects on an injured brain. Interventions for the prevention and treatment of hyperthermia will be explored. Nurses will implement cooling strategies to induce hypothermia, applying interventions to prevent complications, and they will also diagnose hyperthermia, deciding when and if to intervene pharmacologically and therapeutically. These advanced nursing actions will be guided by knowledge and understanding of available evidence. This article presents the pathophysiology of secondary brain injury and how it is affected by both hypothermia and hyperthermia. A review of the research leading up to clinical trials is explored, as well as a discussion of the future of temperature modulation for the brain injury patient. This information will help healthcare providers understand the effect that both hypothermia and hyperthermia have on the acutely injured brain. PMID- 16269895 TI - Prevention of secondary brain injury: targeting technology. AB - Use of technology in the management of the severely brain-injured patient has increased over the past decade and can be confusing and overwhelming to the critical care nurse clinicians who are new to the field of neurology. This article will describe normal physiology and cerebral dynamics and potential abnormal physiology encountered after brain injury. The technology reviewed will include intracranial pressure monitoring, cerebral blood flow monitoring and autoregulation, cerebral oxygen consumption and tissue oxygen monitoring, metabolism, sedation, and temperature monitoring. Integration of appropriate technology into patient management will be discussed using a case study to explore the utility of information at the bedside. Recognizing the difficult task of trying to control secondary injury in our patients is the first step to better outcomes. Implementing the use of technology to mitigate the situation must be done with careful consideration and a team approach to achieve the greatest benefit for the patient. PMID- 16269896 TI - Endovascular coiling for cerebral aneurysms. AB - Aneurysmal subarachnoid hemorrhage is an increasing problem in the United States, affecting approximately 30,000 people every year. Despite advances in the neurosurgical field, approximately 50% of patients die within the first month after hemorrhage. Traditionally, craniotomy with aneurysmal clipping has been employed to manage these patients, but endovascular embolization is moving to the forefront of treatment, particularly for high grade (IV to V) aneurysms. Patient selection is often based on age, aneurysm size, location, characteristics and presentation, and patient hemodynamics. Postprocedure management relies on skilled observers to determine those potential complications that may occur, including vasospasm, rupture, bleeding, or vessel occlusion. Advanced practice nurses have an obligation to be aware not only of the procedure and its management, but also of the potential complications and ongoing care of the patients and families as well. PMID- 16269897 TI - Refractory increased intracranial pressure in severe traumatic brain injury: barbiturate coma and bispectral index monitoring. AB - Patients with severe traumatic brain injury resulting in increased intracranial pressure refractory to first-tier interventions challenge the critical care team. After exhausting these initial interventions, critical care practitioners may utilize barbiturate-induced coma in an attempt to reduce the intracranial pressure. Titrating appropriate levels of barbiturate is imperative. Underdosing the drug may fail to control the intracranial pressure, whereas overdosing may lead to untoward effects such as hypotension and cardiac compromise. Monitoring for a therapeutic level of barbiturate coma includes targeting drug levels and using continuous electroencephalogram monitoring, considered the gold standard. New technology, the Bispectral Index monitor, utilizes electroencephalogram principles to monitor the level of sedation and hypnosis in the critical care environment. This technology is now being considered for targeting appropriate levels of barbiturate coma. PMID- 16269899 TI - Clinical management of the organ donor. AB - There is a critical mismatch between available organs for transplant and acutely or critically ill patients with end-stage organ disease. Patients who may benefit from organ transplantation far outnumber available organs. The causes for this imbalance are multiple. One cause is family refusal to donate. A second cause is nonrecognition or delay in determination of brain death. A third cause is donor loss due to profound cardiopulmonary and metabolic instability consequent to brain-stem herniation and brain death. Family refusal may be addressed by education, public awareness, as well as close attention to social, cultural and ethical issues, and optimal communication with donor families. Brain death may be consequent to traumatic brain injury, ischemic versus hemorrhagic stroke, as well as massive cerebral anoxia/ischemic following cardiac arrest. Nonrecognition or delay in brain death determination may be addressed by clinician education and frequent clinical assessment to detect early stages of brain-stem herniation refractory to aggressive measures for control of intracranial pressure. Donor loss due to profound cardiopulmonary and metabolic instability may be addressed by aggressive, mechanism-based treatment for clinical instability based on affected body system, as well as measures to support metabolic activity at the cellular and tissue level in the brain-dead organ donor. This article explores cerebral physiology related to impending brain death and catastrophic intracranial pressure elevations. In addition, physiologic consequences of brain death are correlated with affected body systems and mechanism-based therapies to support organ function pending transplantation. Ethical/legal issues are explored as related to patient autonomy and optimal family outcomes. Effective family communication, astute clinical assessment, and optimal clinical management of the organ donor are illustrated using a case study approach, highlighting the role of the advanced practice nurse in donor management. PMID- 16269898 TI - Integrating palliative and neurological critical care. AB - The goal of palliative care is to provide the alleviation or reduction of suffering and the support for the best possible quality of life for patients regardless of the stage of the disease. Palliative care can be provided in any patient care setting, including intensive care units. Death in intensive care units is a common occurrence, with literature suggesting that approximately 20% of deaths in the United States occur after a stay in the intensive care unit. Other studies suggest that approximately half of all chronically ill patients who die in a hospital receive care in the intensive care unit within 3 days of their deaths. Critical care nurses who work in neurological intensive care units are at the forefront of integrating palliative and critical care. PMID- 16269900 TI - Neuroscience critical care: the role of the advanced practice nurse in patient safety. AB - The Institute of Medicine report published in 1999 described a healthcare system in which 44,000 to 98,000 patients die each year from preventable medical errors. The healthcare industry has been charged with identifying and ameliorating risks to patients. The advanced practice nurse is in the optimal position to influence the patient care environment and contribute to a culture of patient safety. This article will review the role of the advanced practice nurse in the care of the neuroscience patient in identifying risks unique to this at-risk population. There will be a discussion of risk factors that contribute to errors, with advanced practice nurse-driven, evidence-based solutions. A case presentation of the role of the advanced practice nurse in reducing the incidence of deep vein thrombosis in the craniotomy patients with malignant tumors will be discussed. PMID- 16269905 TI - Norepinephrine transporter: a candidate gene for initial ethanol sensitivity in inbred long-sleep and short-sleep mice. AB - BACKGROUND: Altered noradrenergic neurotransmission is associated with depression and may contribute to drug abuse and alcoholism. Differential initial sensitivity to ethanol is an important predictor of risk for future alcoholism, making the inbred long-sleep (ILS) and inbred short-sleep (ISS) mice a useful model for identifying genes that may contribute to alcoholism. METHODS: In this study, molecular biological, neurochemical, and behavioral approaches were used to test the hypothesis that the norepinephrine transporter (NET) contributes to the differences in ethanol-induced loss of righting reflex (LORR) in ILS and ISS mice. RESULTS: We used these mice to investigate the NET as a candidate gene contributing to this phenotype. The ILS and ISS mice carry different DNA haplotypes for NET, showing eight silent differences between allelic coding regions. Only the ILS haplotype is found in other mouse strains thus far sequenced. Brain regional analyses revealed that ILS mice have 30 to 50% lower [3H]NE uptake, NET binding, and NET mRNA levels than ISS mice. Maximal [3H]NE uptake and NET number were reduced, with no change in affinity, in the ILS mice. These neurobiological changes were associated with significant influences on the behavioral phenotype of these mice, as demonstrated by (1) a differential response in the duration of ethanol-induced LORR in ILS and ISS mice pretreated with a NET inhibitor and (2) increased ethanol-induced LORR in LXS recombinant inbred (RI) strains, homozygous for ILS in the NET chromosomal region (44-47 cM), compared with ISS homozygous strains. CONCLUSIONS: This is the first report to suggest that the NET gene is one of many possible genetic factors influencing ethanol sensitivity in ILS, ISS, and LXS RI mouse strains. PMID- 16269906 TI - Bone mass and strength: phenotypic and genetic relationship to alcohol preference in P/NP and HAD/LAD rats. AB - BACKGROUND: The association between moderate alcohol intake and elevated bone mineral density observed in several epidemiologic studies might result from common genetic pathway regulating both phenotypes. In this study, we determined whether there is a relationship between alcohol preference and high bone mass or strength and whether bone mass-regulating genes segregate during selective breeding of alcohol preferring rats. METHODS: Six different lines of male rats with high or low preference for alcohol consumption were used in this study. The high alcohol preference lines are alcohol-preferring (P), high-alcohol-drinking 1 (HAD1), and high-alcohol-drinking 2 (HAD2), and their corresponding low alcohol preference lines are alcohol-nonpreferring (NP), low-alcohol-drinking 1 (LAD1), and low-alcohol-drinking 2 (LAD2). Bone mass phenotypes were determined using dual energy x-ray absorptiometry (DXA), peripheral quantitative computed tomography (pQCT), and biomechanics in long bones and lumbar vertebrae from rats at 3 and 6 months of age. RESULTS: P rats had significantly higher bone mass and strength compared with NP rats, mainly due to higher cortical bone in long bones and lumbar vertebrae. HAD2 rats also had significantly higher bone mass compared with LAD2 rats, but mostly due to increased trabecular bone leading to increased strength only in lumbar vertebra. Conversely, HAD1 rats had significantly lower bone mass and strength compared with LAD1 rats in long bones. The vertebral bone mass and strength did not differ between HAD1 and LAD1 rats. CONCLUSIONS: This study demonstrated that preference for alcohol consumption had no consistent relationship with high bone mass or strength, as each alcohol-preferring rat line had their unique bone mass phenotypes. However, genes regulating bone mass and strength appear to segregate with alcohol preference genes in P and HAD rat lines, suggesting that alcohol preferring rat lines may be useful for identifying genes that regulate bone mass and structure. PMID- 16269907 TI - The slowpoke gene is necessary for rapid ethanol tolerance in Drosophila. AB - BACKGROUND: Ethanol is one of the most commonly used drugs in the world. We are interested in the compensatory mechanisms used by the nervous system to counter the effects of ethanol intoxication. Recently, the slowpoke BK-type calcium activated potassium channel gene has been shown to be involved in ethanol sensitivity in Caenorhabditis elegans and in rapid tolerance to the anesthetic benzyl alcohol in Drosophila. METHODS: We used Drosophila mutants to investigate the role of slowpoke in rapid tolerance to sedation with ethanol vapor. Rapid tolerance was defined as a reduction in the sedative phase caused by a single previous sedation. The ethanol and water contents of flies were measured to determine if pharmacodynamic changes could account for tolerance. RESULTS: A saturated ethanol air stream caused sedation in <20 min and resulted in rapid tolerance that was apparent 4 hr after sedation. Two independently isolated null mutations in the slowpoke gene eliminated the capacity for tolerance. In addition, a third mutation that blocked expression specifically in the nervous system also blocked rapid tolerance. Water measurements showed that both ethanol and mock sedation caused equivalent dehydration. Furthermore, a single prior exposure to ethanol did not cause a change in the ethanol clearance rate. CONCLUSIONS: Rapid tolerance, measured as a reduction in the duration of sedation, is a pharmacokinetic response to ethanol that does not occur without slowpoke expression in the nervous system in Drosophila. The slowpoke channel must be involved in triggering or producing a homeostatic mechanism that opposes the sedative effects of ethanol. PMID- 16269908 TI - Effects of alcohol on intracellular pH regulators and electromechanical parameters in human myocardium. AB - BACKGROUND: Disturbances in intracellular pH (pHi) of the heart can trigger major changes in the strength and rhythm of the heartbeat. It is well known that two extruders, Na+/H+ exchange (NHE) and Na+/HCO3- symporter (NHS), and a monocarboxylic acid transporter (MCT) are involved in acid-equivalent extruding in the human heart. Drinking alcohol has been proven to affect blood pressure and heart contractility and, sometimes, causes cardiac arrhythmia. To assess the effects of alcohol on pHi regulators and electromechanical parameters, various concentrations of alcohol were superfused into human myocardium in the present study. METHODS: Human atrial myocardium was obtained from hearts of patients undergoing corrective cardiac surgery. Institutional rules for the protection of human subjects were observed. In the whole study, pHi was measured by an epifluorescent, ratiometric microspectrofluorimetry technique with the dye BCECF, while electrophysiological experiments were performed by traditional micropipette. NHE and NHS activities were measured after pHi recovery from intracellular acidosis induced by NH4Cl prepulse, while MCT activity was measured by a lactate adding/removing technique. RESULTS: In pHi experiments, we demonstrated that alcohol could induce a biphasic, concentration-dependent (30 1000 mM) pHi change (i.e., alkalosis after acidosis) in human atrium in HEPES buffered Tyrode solution. To a smaller extent, similar results were found when the superfusate was replaced by HCO3- -buffered Tyrode solution. NHE activity was increased by a moderate concentration of alcohol (30 mM), while it was inhibited in a concentration-dependent manner by higher concentrations of alcohol (>100 mM). On the contrary, 30-1000 mM alcohol increased the activity of NHS in a concentration-dependent manner. Surprisingly, MCT activity was not affected by alcohol. In electromechanical experiments, we found that alcohol (30-1000 mM) had a notable concentration-dependent inhibitory effect on the contractile force, while higher concentrations of alcohol (>100 mM) decreased the action potential amplitude, upstroke velocity, duration of repolarization, and force of contractions in a concentration-dependent way. All these alcohol-induced pHi changes and electromechanical inhibitions were reversible. CONCLUSIONS: To our knowledge, this study provides the first evidence that alcohol can affect pHi in human myocardial tissue by changing the activity of acid extruders (i.e., NHE and NHS). PMID- 16269909 TI - Factors influencing elevated ethanol consumption in adolescent relative to adult rats. AB - BACKGROUND: Many teenagers report frequent alcohol use during the adolescent period. Animal models may be an important tool for exploring factors that contribute to alcohol consumption during this critical period. METHODS: Using a 24-hour, free-access, two-bottle-choice procedure between water and a sweetened solution with or without ethanol in nondeprived rats, the present series of experiments examined the contribution of a variety of contextual and experimental variables (i.e., isolate-housing versus pair-housing, type of sipper tube, caloric value of solution, prior experimental perturbations) on alcohol consumption in both adolescent and adult Sprague-Dawley rats. RESULTS: Ethanol consumption was particularly magnified among adolescent rats using ball bearing containing ball-point (BP) sipper tubes, with this exacerbated intake not due to caloric content of the ethanol solution. Isolation housing for 12 days did not alter ethanol consumption of adolescents relative to their socially housed counterparts while suppressing consumption of isolated adults. An examination of differences in the relative magnitude of adolescent ethanol consumption across experiments in this series revealed that ethanol intake among adolescents was elevated not only by the inclusion of BP sipper tubes but also by staggering the timing of isolate housing relative to the presentation of the novel ethanol solution. CONCLUSIONS: The results of this experimental series demonstrate that adolescent animals consume significantly more ethanol than adult animals under a variety of home cage continuous-access circumstances, with the relatively greater intake of adolescents further magnified by a number of test conditions. Subtle experimental details often thought to be innocuous can have a substantial impact on overall amount of voluntary ethanol consumption observed in both adolescent and adult animals. PMID- 16269911 TI - A microdialysis profile of Met-enkephalin release in the rat nucleus accumbens following alcohol administration. AB - BACKGROUND: Pharmacological studies have implicated the endogenous opioid system in mediating alcohol intake. Other evidence has shown that alcohol administration can influence opioid activity. In this regard, the majority of studies have concentrated on endorphinergic systems, whereas other opioid systems have been granted comparably less attention. This is the case despite some compelling evidence that has implicated enkephalinergic peptide systems, particularly Met enkephalin, in mediating alcohol preference. The aim of the present study was to investigate the effect of alcohol administration on extracellular levels of Met enkephalin in the rat nucleus accumbens--a brain region that plays a significant role in the processes underlying reinforcement and stress. METHODS: Male Sprague Dawley rats were implanted with a microdialysis probe aimed at the shell region of the nucleus accumbens. Artificial cerebrospinal fluid was pumped at a rate of 1.75 mul/min in awake and freely moving rats and dialysates were collected at 30 minute intervals. After several baseline collections, rats were injected intraperitoneally with either physiological saline or one of four doses of alcohol: 0.8, 1.6, 2.4, or 3.2 g/kg ethanol body weight. The levels of Met enkephalin in the dialysates were analyzed with solid-phase radioimmunoassay. RESULTS: Within the first 30 minutes of administration, an alcohol dose of 1.6 g/kg caused a significant and prolonged elevation in the extracellular levels of Met-enkephalin. Alcohol did not have a major effect on the release of Met enkephalin at any other dose. CONCLUSIONS: In this experiment, only a moderate dose of alcohol was capable of stimulating Met-enkephalin release in the nucleus accumbens. Enkephalins may modulate local neurotransmitter release by binding to presynaptic Delta-opioid receptors, or, they may inhibit effector cells by binding to postsynaptic Delta- or mu-opioid receptors. This may be one of multiple neurological mechanisms that modulate alcohol-drinking behavior. PMID- 16269910 TI - Sensitivity and tolerance to autonomic effects of ethanol in adolescent and adult rats during repeated vapor inhalation sessions. AB - BACKGROUND: It is during adolescence that most drinkers initiate ethanol intake, with some of this use being excessive. One possible contributor to the increased ethanol consumption often seen during adolescence in humans and in various animal models is age differences in ethanol sensitivity and tolerance. The present study examined the impact of age on ethanol-related alterations in the autonomic nervous system. METHODS: Sensitivity to the initial ethanol challenge and chronic tolerance as well as acute and protracted withdrawal-like phenomena were assessed in male adolescent and adult Sprague-Dawley rats, using implanted telemetry probes with ethanol delivered via vapor inhalation. RESULTS: Both ages showed similar ethanol-induced tachycardia and activity suppression; however, adolescents were found to be more sensitive than adults to the hypothermic effect of ethanol, data opposite other results from our laboratory and elsewhere using intragastric intubations or intraperitoneal administrations of ethanol. Although little tolerance to ethanol's tachycardic or activity suppressant effects was seen after repeated ethanol inhalation sessions, chronic tolerance to ethanol's hypothermic effect developed faster in adults than in adolescents. A withdrawal like syndrome, characterized by bradycardia and hypoactivity, typically emerged during the dark phase of the diurnal cycle after ethanol vapor exposure sessions. These effects were observed in animals of both ages, with the bradycardic effect more pronounced in adolescents. CONCLUSIONS: In contrast to results indicating that adolescents may be less sensitive than adults to ethanol's hypothermic effect when ethanol is administered via bolus injection/intubation, adolescents appear more sensitive and develop tolerance to ethanol's hypothermic effects more slowly than adults when ethanol is administered at a more moderate rate via vapor inhalation. PMID- 16269912 TI - Enhanced prepulse inhibition following adolescent ethanol exposure in Sprague Dawley rats. AB - OBJECTIVES: Recent studies have demonstrated that ethanol exposure differentially affects adolescents and adults. The current studies were designed to compare the effects of 2-week exposure to ethanol during adolescence or adulthood on the acoustic startle response (ASR) and prepulse inhibition (PPI) METHODS: Male Sprague-Dawley rats were exposed to ethanol vapor 12 hr/d (on from 6 pm to 6 am) for 14 days during adolescence or adulthood. Six days after the cessation of ethanol vapor exposure, the ASR and PPI were assessed. RESULTS: During ethanol treatment, overall blood alcohol levels averaged 230 to 250 mg/dl in the adolescent and adult treatment groups. Assessment of the ASR revealed that latency to startle was more rapid in adolescents than in adults, but ASR latency was not altered by ethanol exposure. In addition, ASR magnitude was lower in adolescents and was decreased in ethanol-exposed rats on startle trials. Ethanol exposure significantly enhanced PPI, but only after adolescent exposure CONCLUSIONS: These data further demonstrate a differential sensitivity of adolescents and adults to the effects of ethanol exposure. Specifically, a 2-week period of ethanol exposure during adolescence selectively enhanced PPI, a neurobehavioral index of sensorimotor gating. However, ASR magnitude was decreased by ethanol exposure regardless of age. On the basis of previous studies, the effects of ethanol exposure on PPI data could indicate that adolescent rats exposed to ethanol are more likely to exhibit behavioral inflexibility and that ethanol exposure acts as a more potent physical stressor in adolescent rats. PMID- 16269913 TI - Role of parvalbumin in estrogen protection from ethanol withdrawal syndrome. AB - BACKGROUND: Parvalbumin (PA) is a calcium-binding protein that has been implicated in protecting neurons from hyperexcitability by sequestering intracellular calcium. This study examined whether ethanol exposure and/or ethanol withdrawal (EW) alter the levels of PA in a manner that is protected by 17beta-estradiol (E2). METHODS: Ovariectomized rats implanted with E2 (EW/E2) or oil pellets (EW/Oil) received chronic ethanol (7.5% w/v, 5 weeks) or control dextrin (Dex/Oil and Dex/E2) diets. At 0 hr, 24 hr, and 2 weeks of EW, three brain areas (the cerebellum, hippocampus, and cortex) were prepared for immunoblotting and immunohistological assessment of PA. RESULTS: At 24 hr of EW, the EW/Oil group showed reduced levels of PA protein and PA-positive neurons in the cerebellum and hippocampus compared with the dextrin control and the EW/E2 groups. At 2 weeks of EW, the reduced levels of PA persisted in the cerebellum but recovered toward the control levels in the hippocampus. The cortex showed no change in PA levels in any of the treatment groups. When tested at 24 hr of EW, the magnitude of EW signs inversely correlated with the levels of PA in the cerebellum and hippocampus. Ethanol exposure itself did not affect PA levels. CONCLUSION: These data suggest that EW, rather than ethanol exposure, reduces PA levels in a manner that is brain region specific and that is protected by estrogen. Disturbed PA homeostasis is hypothesized to play a role in the hyperexcitability of EW signs. PMID- 16269915 TI - Rat breathalyzer. AB - BACKGROUND: The purpose of this study was to characterize the estimation of blood alcohol (ethanol) concentration (BAC) in laboratory rats by measuring ethanol concentration in breath (BrAC) using a specialized apparatus in combination with a gas chromatography system. METHODS: The apparatus consisted of a body chamber, a plastic cylinder, from which the head of the rat protruded, a head chamber, and a water-jacketed cylinder, in which the rat's head was placed while the breath sample was collected. The breath sample was withdrawn from the head chamber through a sample loop by a Minipuls pump and then injected directly into the gas chromatography system that was equipped with a flame ionization detector for the quantification of ethanol. For these experiments, Lewis rats were catheterized 1 week before the commencement of the experiments so that blood samples were collected at exactly the same time as the breath samples. RESULTS: Our results show that Lewis rats can be trained to enter and be secured in the body chamber and that they appear to be comfortable for periods as long as 150 min. The profiles of the pharmacokinetic curves for BrAC and BAC were essentially identical. Cmax for BrAC and BAC at 8 min after the intraperitoneal injection of ethanol was directly proportional to the doses of ethanol. The ratio of BrAC expressed as peak area to BAC (expressed as mM) was calculated to be 3282. This conversion factor can be used to directly estimate the BAC from the BrAC. CONCLUSIONS: The principal conclusion of this study was that the rat breathalyzer is an accurate and convenient laboratory method to estimate BAC in a noninvasive manner. This procedure will be particularly useful for studies requiring repeated assessment of alcohol levels. PMID- 16269916 TI - Paths of entry into alcoholics anonymous: consequences for participation and remission. AB - BACKGROUND: This study compared individuals with alcohol use disorders who, in the first year after initiating help-seeking, entered Alcoholics Anonymous (AA) only, entered professional treatment and AA together, or entered professional treatment only METHODS: A sample of initially untreated individuals (N = 362) was surveyed at baseline and 1 year, 3 years, 8 years, and 16 years later. At each contact point, participants described their participation in AA and treatment and their current alcohol-related functioning. They also described their reasons for entering AA and/or treatment and the perceived benefits of these sources of help RESULTS: Compared with individuals who initially participated only in treatment but later entered AA, those who entered treatment and AA together participated in AA longer and more frequently and were more likely to achieve remission. Among individuals who initially participated only in AA, those who later entered treatment had poorer remission outcomes than those who did not enter treatment. Longer duration of participation in AA was associated with a higher likelihood of remission at all four follow-ups; individuals who dropped out of AA were more likely to relapse or remain nonremitted. CONCLUSIONS: Compared with individuals who participated only in professional treatment in the first year after they initiated help-seeking, individuals who participated in both treatment and AA were more likely to achieve remission. Individuals who entered treatment but delayed participation in AA did not appear to obtain any additional benefit from AA. PMID- 16269914 TI - Neonatal alcohol exposure permanently disrupts the circadian properties and photic entrainment of the activity rhythm in adult rats. AB - BACKGROUND: Alcohol exposure during the period of rapid brain development produces structural damage in different brain regions, including the suprachiasmatic nucleus (SCN), that may have permanent neurobehavioral consequences. Thus, this study examined the long-term effects of neonatal alcohol exposure on circadian behavioral activity in adult rats. METHODS: Artificially reared Sprague-Dawley rat pups were exposed to alcohol (EtOH; 4.5 g/kg/day) or isocaloric milk formula (gastrostomy control; GC) on postnatal days 4-9. At 2 months of age, rats from the EtOH, GC, and suckle control (SC) groups were housed individually, and properties of the circadian rhythm in wheel-running behavior were continuously analyzed during exposure to a 12-hr light:12-hr dark photoperiod (LD 12:12) or constant darkness (DD). RESULTS: Neonatal alcohol exposure had distinctive effects on the rhythmic properties and quantitative parameters of adult wheel-running behavior. EtOH-treated animals were distinguished by unstable and altered entrainment to LD 12:12 such that their daily onsets of activity were highly variable and occurred at earlier times relative to control animals. In DD, circadian regulation of wheel-running behavior was altered by neonatal alcohol exposure such that the free-running period of the activity rhythm was shorter in EtOH-exposed rats than in control animals. Total amount of daily wheel-running activity in EtOH-treated rats was greater than that observed in the SC group. In addition, the circadian activity patterns of EtOH-exposed rats were fragmented such that the duration of the active phase and the number of activity bouts per day were increased. CONCLUSIONS: These data indicate that neonatal alcohol exposure produces permanent changes in the circadian regulation of the rat activity rhythm and its entrainment to LD cycles. These long-term alterations in circadian behavior, along with the developmental alcohol-induced changes in SCN endogenous rhythmicity, may have important implications in clinical sleep-wake disturbances observed in neonates, children, and adults exposed to alcohol in utero. PMID- 16269918 TI - Mean corpuscular volume, alcohol flushing, and the predicted risk of squamous cell carcinoma of the esophagus in cancer-free Japanese men. AB - BACKGROUND: Because some of the causes of increased mean corpuscular volume (MCV) and esophageal squamous cell carcinoma (ESCC), including alcoholism, acetaldehyde exposure, smoking, and poor nutrition are common to both, macrocytosis has been used as a predictor of early ESCC in Japanese alcoholics. We examined whether this was also true in the Japanese general population. METHODS: This study compared the MCV of 522 cancer-free Japanese men with his risk of ESCC as defined using drinking, smoking, dietary habits and aldehyde dehydrogenase-2 (ALDH2) genotype in a previous case-control study of ESCC involving them as control subjects. RESULTS: MCV was significantly correlated with ESCC risk predicted by drinking combined with ALDH2 genotype, smoking, or fruit intake. Men at higher risk of ESCC were more frequent in the groups with higher MCV (p < 0.0001 for trend). The replies to a questionnaire about facial flushing in response to alcohol showed that the trend was more prominent in men with current/former flushing, a surrogate marker for inactive ALDH2, than in men with no flushing (p < 0.0001). In comparison with the mean risk of men with MCV < or = 93 fl (lowest quartile), that of current/former flushing men with MCV > or = 99 fl (highest quartile) was 6.35-fold higher, whereas that of never-flushing men with MCV > or = 99 fl was 2.50-fold higher. The sensitivity and specificity of the combination of moderate-to-heavy drinking and either MCV > or = 99 fl or current/former flushing, either 30+ pack-years or MCV > or = 99 fl or either 30+ pack-years or current/former flushing for detection of high-risk persons ranking in the top 10%, was 85% and 84%, 94% and 76%, or 98% and 77%, respectively. CONCLUSIONS: MCV and alcohol flushing might be used to better select candidates to screen this high-mortality-rate cancer not only in alcoholics but also in nonalcoholic Japanese men. PMID- 16269917 TI - Relationship of age of first drink to child behavioral problems and family psychopathology. AB - BACKGROUND: Studies have implicated a wide variety of variables as being associated with an early age of first drink (AFD). AFD in turn has been associated with a variety of negative outcomes in adolescence and early adulthood. This study is designed to quantify the contributions of these antecedent variables to prediction of AFD; in particular it will carefully examine the involvement of variables in four areas (child characteristics, family demographics, family psychopathology, and child behavior problems). METHODS: Using data from a multicenter study on alcoholism, we first investigated the differences between two groups of children (ages 7 to 17 years), one from families heavily loaded for alcohol dependence and the other from population controls. Second, a multidomain, multistep regression model using child characteristics, family demographics, family psychopathology, and child behavior problems was performed to determine significant contributors to predicted AFD. RESULTS: Five variables initially contributed to the prediction of AFD. These included gender, age at interview, the number of adult sibs with alcohol dependence, being held back a year in school, and conduct scale score. However, the number of conduct symptoms appeared to contain the contributions of gender and being held back a grade in school, and these two variables were subsequent removed from the model. The remaining three variables explained 45% of the model variance; age at interview accounted for 38.3%, conduct scale score accounted for 6.2%, and the number of alcohol-dependent adult sibs accounted for 0.5%. No family history measures of alcohol dependence or antisocial personality disorder were contributory to the prediction model for AFD. CONCLUSIONS: Both the "number of conduct symptoms" and the "number of adult sibs with alcohol dependence" are inversely associated with predicted AFD. The latter variable appears marginally predictive of AFD and suggests a condition in which the child's household, regardless of strength of family history of AD (or antisocial personality disorder), appears conducive to early drinking. Thus, child and environmental factors are stronger predictors of age of first drink than family history. PMID- 16269919 TI - The composition of surrogate alcohols consumed in Russia. AB - BACKGROUND: In the course of a case-control study examining determinants of premature death among working age men, it became clear that a significant percentage of the population (7.3%) were drinking a variety of surrogate alcohol products (products not legally sold for consumption). In this population, where there is a high death rate from alcohol-related causes, including acute alcohol poisoning, it was important to know what these products contained. METHODS: The identity of products being consumed was identified from the survey of controls. Representative samples were obtained and subjected to analysis using gas chromatography and mass spectrometry to determine their composition. RESULTS: Three broad groups of product were identified: samogon (home-produced spirits); medicinal compounds; and other spirits (mainly sold as aftershaves). Commercially produced vodkas were used for comparison. Samogon contained lower quantities of ethanol than vodka [mean, 39 vs. 44 volumetric percentage (v/v%), respectively] but in addition contained certain toxic long-chain alcohols. Medicinal compounds contained only ethanol, at a higher concentration that vodka (mean, 66 v/v%), while the other spirits, which were also essentially pure ethanol, contained a mean of 94 v/v%. CONCLUSIONS: A significant number of Russian men are drinking products that have either very high concentrations of ethanol or contaminants known to be toxic. These products are untaxed and thus much less expensive than vodka. There is an urgent need for policy responses that target their production and consumption. PMID- 16269920 TI - Alcohol-induced endothelial changes are associated with oxidative stress and are rapidly reversed after withdrawal. AB - BACKGROUND: Although heavy alcohol drinkers are at an increased risk of developing cardiovascular events, moderate alcohol intake is associated with reduced incidence of cardiovascular death. This paradox might reflect a dose related effect of different alcohol intakes on endothelial function and this, in turn, might depend on changes in oxidative stress. METHODS: We tested the effects of alcohol withdrawal in heavy alcohol consumers and compared the plasma levels of endothelin-1, nitric oxide, plasminogen activator inhibitor-1, von Willebrand factor, malondialdehyde, and intracellular glutathione with those of alcoholics that did not modify their alcohol intake and teetotalers. In human endothelial cells that had been cultured for 2 weeks in the presence of different concentrations of ethanol, we assessed the same parameters after withdrawal of ethanol exposure. RESULTS: Alcohol increased the levels of endothelin-1, nitric oxide, and plasminogen activator inhibitor-1 and decreased the levels of von Willebrand factor both in vivo and in vitro. These changes were dose dependent, rapidly reversed after withdrawal of exposure, and associated with the presence of increased oxidative stress as indicated by increased levels of both malondialdehyde and intracellular glutathione. Blockade of oxidative stress by incubation of endothelial cells in the presence of oxidants' scavengers prevented the alcohol-induced functional modifications of the endothelium. CONCLUSIONS: Alcohol affects endothelial function with an effect that is mediated by an activated oxidative stress and is rapidly reversed after withdrawal. Dose-related endothelial responses to different alcohol intakes might translate in either vascular protection or vascular damage. PMID- 16269922 TI - Alcohol consumption and platelet activation and aggregation among women and men: the Framingham Offspring Study. AB - BACKGROUND: Alcohol intake has been associated with lower platelet activity; however, few large-scale studies have included women, and to our knowledge, the relationship of alcohol intake with measures of platelet activation has not been studied. METHODS: We performed a cross-sectional analysis of adults free of cardiovascular disease enrolled in the Framingham Offspring Study. Study physicians assessed alcohol consumption with a standardized questionnaire. We measured platelet activation in response to 1 and 5 microm of adenosine diphosphate (ADP) with a P-selectin assay among 1037 participants and platelet aggregability in response to ADP, epinephrine, and collagen among 2013 participants. RESULTS: Alcohol consumption was inversely associated with P selectin expression in response to 1 microm ADP (p = 0.007) and 5 microm ADP (p = 0.02) among men but not women. Alcohol consumption was also inversely associated with platelet aggregation induced by ADP among both women (p = 0.04) and men (p trend = 0.008) and by epinephrine among men (p = 0.03) CONCLUSIONS: Alcohol consumption is inversely associated with both platelet activation and aggregation, particularly in men. Additional research is needed to determine whether these findings contribute to the contrasting associations of alcohol consumption with risk of thrombotic and hemorrhagic cardiovascular events. PMID- 16269923 TI - Is herbal "root" effective for reducing alcohol drinking? PMID- 16269921 TI - Opposing effects of chronic alcohol consumption on hepatic gluconeogenesis for female versus male rats. AB - BACKGROUND: The impact of chronic alcohol consumption on hepatic gluconeogenesis (HGN) between males and females is unknown. To determine the effects of chronic alcohol consumption (8 weeks) on HGN, the isolated liver perfusion technique was used on 24-hr-fasted male and female Wistar rats. METHODS: After surgical isolation, livers were perfused (single pass) for 30 min with Krebs-Henseleit bicarbonate buffer and fresh bovine erythrocytes with no added substrate (washout period). After the washout period, livers were perfused with lactate (10 mM) and [U-14C]lactate (15,000 dpm/ml) using the recirculation method. RESULTS: There was no significant difference in HGN between males and females fed the control diet. In contrast, the females chronically fed the ethanol diet (FE) had significantly lower HGN rates (2.73 +/- 0.37 micromol/min x g liver protein(-1)), whereas males fed the ethanol diet (ME) had significantly higher HGN rates (4.99 +/- 0.45 micromol/min x g liver protein(-1)) than controls (3.83 +/- 0.34 micromol/min x g liver protein(-1)). Concomitant decreases were also observed for both 14C-lactate incorporation into 14C-glucose and rates of lactate uptake for FE, while corresponding increases were observed for 14C-lactate incorporation into 14C glucose for ME. The livers from ME were able to convert a greater percentage of the lactate into glucose, resulting in the elevation in gluconeogenic capacity. CONCLUSION: Chronic alcohol consumption lowers the hepatic gluconeogenic capacity from lactate in females and elevates HGN in males. PMID- 16269925 TI - Back to the future: testing in disseminated intravascular coagulation. AB - Following on from the first clinical observations on disseminated intravascular coagulation (DIC) in the nineteenth century, the dawn of laboratory testing for DIC began with the availability of assays that characterized the extrinsic and intrinsic pathways of coagulation. Markedly increased clotting times were the hallmark of DIC. As the understanding of the biochemistry of haemostasis and thrombosis improved, new tests were developed based on the molecular players that participate in the process. However, many are non-specific for DIC and/or are unwieldy in performance to keep apace with the demands of the acute clinical setting. The renewed emphasis in DIC for the modern laboratory of the twenty first century has seen a return to the simple, rapid and practical global tests of coagulation within scoring systems that also capture the pathophysiological continuum by trend analysis. Additionally, new technologies based on these simple tests of coagulation hold promise in also indicating the in vivo interplay between coagulation and inflammation during DIC. PMID- 16269926 TI - Cancer procoagulant in patients with adenocarcinomas. AB - Cancer procoagulant (CP) is a cysteine proteinase that may be produced by malignant and foetal tissue. The possible role of CP in the pathogenesis of cancer-related thrombosis has been suggested recently. The purpose of the study was to evaluate coagulation prothrombotic markers and their relation to CP concentration in the blood of patients with gastrointestinal adenocarcinomas (GIAC). The study group consisted of 45 patients with confirmed diagnosis of adenocarcinoma (stomach, 18 patients; colon, 27 patients) and without evident metastatic disease. In 24 patients further observation showed metastases. The control group for CP was composed of 10 healthy subjects. Blood samples were drawn on the admission day, before any treatment. Among 45 patients with GIAC, deep venous thrombosis was observed in two (4.4%). In all patients the CP activity in the serum was found, and the mean CP activity shortened the coagulation time almost three times compared with the healthy control group. Also, the mean thrombin-antithrombin complex concentration was above the normal range. A significant elevation of the mean prothrombin fragment 1+2 plasma content in this group of patients was noticed. Despite these observations, CP remained within the normal range and did not correlate with thrombin-antithrombin complex or prothrombin fragment 1+2 plasma concentrations. A positive correlation was observed between serum CP and fibrinogen concentration, and a negative correlation between CP and free protein S plasma content (P = 0.04 and P = 0.025, respectively). A negative correlation between activated protein C resistance ratio and protein C activity in the plasma was confirmed. Protein C activity in the plasma showed a correlation with free protein S plasma content. Analysis of factors influencing the activated partial thromboplastin time revealed the presence of antiphospholipid antibodies in seven persons from the study group (in three cases of IgG and in four cases of IgM class). Our data suggest that CP is a minor risk factor for deep venous thrombosis in GIAC patients. To confirm this, however, the number of patients and controls should be larger. After 3 years of observation, the follow-up in 10 living GIAC patients showed nobody with thromboembolic disease. PMID- 16269927 TI - Control of bleeding in children with Dengue hemorrhagic fever using recombinant activated factor VII: a randomized, double-blind, placebo-controlled study. AB - OBJECTIVES: We evaluated the efficacy and safety of recombinant activated factor VII (rFVIIa) in children aged < 18 years old with grade II or grade III Dengue hemorrhagic fever (DHF) who required blood component therapy for controlling bleeding episodes. STUDY DESIGN: Patients were randomized to the rFVIIa group or placebo group in a ratio of 2:1. rFVIIa or placebo (100 microg/kg body weight) was given by intravenous bolus injection. When bleeding was not effectively controlled, a second dose of rFVIIa or placebo (100 microg/kg) was given 30 min after the first dose. RESULTS: Nine and 16 patients received placebo and rFVIIa, respectively. The demographics, bleeding manifestations and grade of DHF were similar for the rFVIIa and placebo groups. Apart from petechiae and ecchymosis, one to four additional bleeding sites were found in each patient, including hematemesis (n = 15), epistaxis (n = 14), gum bleeding (n = 12), melena (n = 7), hypermenorrhea (n = 4), hematochezia (n = 2) and hematuria (n = 2). The mean total dose of rFVIIa (138.4 +/- 50.9 microg/kg) and placebo (145.4 +/- 53.7 microg/kg) were comparable. The efficacy of bleeding control at 2 h after the first dose was completely ceased (rFVIIa 75.0% versus placebo 44.4%), decreased (rFVIIa 18.7% versus placebo 11.2%), and unchanged or worsened (rFVIIa 6.3% versus placebo 44.4%). Some patients with active bleeding received platelet concentrates 3-12 h after the first dose of rFVIIa or placebo. The subsequent efficacy of bleeding control at 6, 12 and 24 h was comparable between the two groups. The cumulative use of red blood cells (rFVIIa 31.3% versus placebo 33.3%) and plasma (rFVIIa 25% versus placebo 22%) during the 24-h period was not significantly different between the two groups. In contrast, platelet concentrate requirement in the rFVIIa group (6.3%) was lower than the placebo (33.3%). No clinical evidence of thromboembolic complications or mortality as a result of bleeding was observed. CONCLUSION: rFVIIa appears to be a useful adjunctive treatment to blood component transfusion for controlling active bleeding in children with DHF especially when platelet concentrate is not readily available. PMID- 16269928 TI - Platelet hyper-function in acute coronary syndromes. AB - Previous studies have demonstrated shortened bleeding times in patients with acute coronary syndromes, especially in myocardial infarction (MI). In this study we have investigated platelet hyper-function using the PFA-100 with collagen/adenosine diphosphate and collagen/epinephrine cartridges in 78 patients presenting with acute chest pain. Patients were classified into MI, unstable angina (UA) and non-specific chest pain. All patients received 300 mg aspirin (ASA) more than 2 h before blood samples were collected. Twenty healthy normal subjects were also tested before and 2 h after 300 mg ASA (n = 10). The collagen/adenosine diphosphate closure time was significantly shorter in MI patients (median, 71 s; P = 0.0237) but not in UA patients (median, 81 s; P > 0.05) compared with normal subjects (median, 92.5 s). The collagen/epinephrine closure times were significantly longer in UA patients (median, 233 s) than in untreated controls (median, 125 s; P < 0.0001), as expected, but there was no difference in MI patients (median, 149.24 s; P > 0.05), suggesting that the MI patients were not all responding to ASA. Analysis of a subset of the apparent ASA non-responders (n = 5) by platelet aggregation demonstrated that this was not related to failure of ASA to block cyclo-oxygenase activity. Von Willebrand factor levels were significantly elevated in both UA and MI patients compared with normal subjects (mean, 175.5 and 248.9 versus 89.1 s; P < 0.0001 and P < 0.0001, respectively) and were also significantly higher in the MI group compared with the UA group (P < 0.05). There is evidence for platelet hyper-function and elevated von Willebrand factor levels in the MI group that could explain their decreased responsiveness to ASA on the collagen/epinephrine cartridge. PMID- 16269929 TI - No impact of factor IX Ala-10 mutations in acenocoumarol-treated southern Europeans. AB - Increased risk of bleeding during oral anticoagulant (OA) treatment may be related to mutations in the precursor of coagulation factor IX. Missense mutations at Ala-10 (Ala-10Thre and Ala-10Val) in the factor IX propeptide lead to impaired carboxylation of factor IX. When patients carrying these mutations are treated with coumarins, functional factor IX levels decrease significantly, leading to an excessively prolonged activated partial thromboplastin time (aPTT) and an increased bleeding risk. Mutations at Ala-10 have been described in northern-European patients, but it is not known whether geographical differences play a role in the prevalence of these mutations. We aimed to analyze the prevalence of mutations at Ala-10 in factor IX in southern-European patients on OA treatment. Patients attending the Oral Anticoagulant Unit of the Hospital Clinic were prospectively included. The aPTT was determined at their normal International Normalized Ratio (INR) control. When the aPTT was excessively prolonged for the INR value, determination of factor IX and genotyping for Ala-10 mutations was performed. A total of 2360 patients were included (1289 men, 1071 women; mean age, 70.5 +/- 12.1 years). Twenty-four patients (16 men, eight women; mean age, 61.0 +/- 16.2 years) had an aPTT over that expected for the INR. The mean aPTT was 69.6 +/- 16.2 s. Only one patient presented with a factor IX level lower than 10%. None of the 24 patients carried mutations at Ala-10. Mutations at Ala-10 in factor IX were non-existent in the southern-European population analyzed, and thus, do not represent a relevant cause of bleeding during OA treatment. PMID- 16269930 TI - Phlegmasia cerulea dolens as a complication of deep vein thrombosis in a man with primary antiphospholipid syndrome. AB - We report here the occurrence of phlegmasia cerulea dolens of both lower extremities as a complication of massive deep vein thrombosis in a 37-year-old man with primary antiphospholipid syndrome. The patient had no history of thromboembolic events or any comorbidities. Clinical and laboratory evaluation revealed a strong positive lupus anticoagulant as the unique thrombotic risk factor. In the literature, there is a single case of phlegmasia cerulea dolens as a complication of deep vein thrombosis in an obese woman with a history of thrombosis. We show that phlegmasia may occur in a previously healthy man with positive lupus anticoagulant. PMID- 16269931 TI - Efficacy of Haemate-P as prophylaxis of recurrent bleeding in a patient with type 2B von Willebrand's disease. AB - We report the case of a 58-year-old woman with type 2B von Willebrand's disease who received regular infusions of the factor VIII/von Willebrand factor concentrate Haemate-P (2000 IU every other day) for 4 months to prevent recurrent bleeding from a malignant esophageal ulcer. Prophylaxis with Haemate-P was shown to be safe and effective in this patient, thus enabling completion of radiochemotherapy. PMID- 16269932 TI - Successful use of recombinant activated factor VII in the treatment of vitreous haemorrhage: a report of seven cases. AB - Vitreous haemorrhage poses a serious threat to vision if untreated. Therapeutic options remain scarce and surgical intervention to resolve persistent bleeding is associated with risks that may further compromise vision. We report the use of recombinant activated factor VII (rFVIIa) in seven patients (six men, one woman; age, 30-65 years) with vitreous haemorrhage and severe reduction in visual acuity caused by trauma (n = 4) or proliferative diabetic retinopathy (n = 3). Initial doses ranged from 60 to 140 microg/kg; most patients received maintenance therapy with 20-60 microg/kg for at least 3 days. One patient received rFVIIa treatment for only 24 h and suffered a re-bleed, controlled successfully with further rFVIIa therapy. Five patients responded well to rFVIIa treatment, with reduced symptoms and improvements in visual acuity. Late presentation several days after trauma or symptom onset may have contributed to poor outcomes in the two patients who failed to respond to rFVIIa therapy. No adverse events were observed. An initial dose of rFVIIa 60-140 microg/kg, followed by 20-60 microg/kg repeated at 8-h intervals for 3-5 days, appears to be an effective therapeutic option for vitreous haemorrhage. However, further studies of rFVIIa use in this indication are warranted. PMID- 16269933 TI - A new optimized method for the determination of fibrin clot permeability. AB - The measurement of fibrin clot permeability is important in the investigation of fibrin clot characteristics. A major disadvantage of the standard measurement techniques is that they are time consuming as they require constant adjustments and monitoring. Presented here is a new method that is easy to perform and does not need continuous adjustments. In addition, the supervision required during the measurement is greatly minimized. When performing the measurement in quadruple, the intra-run coefficient of variation is 11.7%, which is comparable with other reported methods. A (5.1-13.1) x 10(-9) cm reference interval of the permeability coefficient (Ks) among 22 healthy volunteers was established. PMID- 16269934 TI - Identifying hypocoagulable states with a modified global assay of overall haemostasis potential in plasma. AB - To test the sensitivity of the global assay of overall haemostasis potential (OHP) in detecting hypocoagulation, the OHP was assayed in plasma containing exogenous thrombin (0.04 IU/ml), tissue-plasminogen activator (330 ng/ml), Ca and a platelet reagent. Commercial plasmas with factor II, V, VIII, IX, X, XI, XII or VII deficiency were mixed with normal plasma in different proportions to imitate different severities. Samples from patients with haemophilia and factor XII deficiency were also examined. No clot was found in the absence of factor II/factor X, indicating that the tiny dose of thrombin worked solely as a trigger for the intrinsic pathway activation. Changed levels of the investigated coagulants, apart from factor XII, influenced the outcome. OHPs were decreased in patients with haemophilia but were unchanged or even increased in those with factor XII deficiency. This modified OHP method may therefore be useful for estimating the bleeding tendency in haemophilic patients and to find suitable doses and intervals for prophylactic treatment. It may also be of use in investigations of the effect of antifibrinolytic drugs as well as for identifying a thrombotic tendency in patients with factor XII deficiency. For detection of other coagulation factor deficiencies, our investigations with the commercial plasmas suggest that the OHP assay is also valuable, especially when the intrinsic pathway of the coagulation cascade is impaired. PMID- 16269935 TI - Cross-laboratory audit of normal reference ranges and assessment of ABO blood group, gender and age on detected levels of plasma coagulation factors. AB - We have assessed the influence of the ABO blood group, gender and age on plasma levels of the coagulation factors II, V, VII, VIII, IX, X, XI and XII as part of a quality audit of laboratory activities. There was no statistically significant difference in gender donor age (total normal donors: n = 406, mean/median age = 46.0/47.0 years, range = 16-77 years; females: n = 177, mean/median age = 44.7/46.0 years, range = 16-75 years; males: n = 229, mean/median age = 47.0/48.0 years, range = 17-77 years). With increasing age, we observed small but statistically significant rises (linear correlation; P < 0.01 for all parameters) in factors V, VII, VIII, IX and XI. With gender, we observed higher levels (P < 0.05) in females for factors II, VII, X, IX, XI and XII. With the ABO group, we observed lower levels in the O group (versus non-O group; P < 0.05) for factors VIII, IX and XII. We could therefore define differing normal reference ranges based on the differing study data. Study findings are compared with previously published literature, and this has identified a wide diversity in normal reference ranges both between different factors and between different studies. Finally, we also performed a cross-laboratory audit of peer laboratory practice and similarly show a wide diversity in normal reference ranges used between different laboratories. PMID- 16269936 TI - Cleavage of von Willebrand factor by ADAMTS-13 in vitro: effect of temperature and barium ions on the proteolysis kinetics. AB - The multimeric size of von Willebrand factor (VWF) is regulated by the specific cleaving metalloprotease, ADAMTS-13. Laboratory assays for ADAMTS-13 are useful for identifying severe protease-deficient activity. ADAMTS-13 activity is currently assayed by prolonged dialysis of plasma at 37 degrees C in low-ionic strength denaturing buffer. We investigated the effect of temperature and divalent cation supplementation on the kinetics of VWF proteolysis by ADAMTS-13 in vitro. Proteolysis was monitored for 24 h at 37, 22, and 4 degrees C, in the presence or absence of barium ions, by measuring the binding of VWF to collagen. Complete VWF proteolysis was observed at 37 degrees C in the presence of BaCl2, while about 25% VWF still bound to collagen when BaCl2 supplementation was omitted. Proteolysis kinetics at 22 and 4 degrees C was slower but complete, even in the absence of added barium. A subphysiological temperature might influence the proteolysis kinetics by determining minor variations of the ADAMTS-13 structure, or further modification of the VWF substrate. We describe a simple procedure to analyse the kinetics of VWF proteolysis that is suitable for routine diagnostic use. Furthermore, we offer new insight into the biochemistry of ADAMTS 13. PMID- 16269937 TI - An evaluation of the procoagulant action of recombinant activated factor VII in cord whole blood versus adult whole blood using thromboelastography. AB - Recombinant activated factor VII (rFVIIa) has been reported to be effective in adult patients in various clinical situations and might be beneficial in neonates with bleeding tendency. In the present study we compared the procoagulant action of increasing amounts of rFVIIa in both cord whole blood and adult whole blood with respect to changes in the values of the clotting time, clot formation time, and maximum clot firmness by means of thromboelastography. Thromboelastography allows evaluation of the effects of rFVIIa on haemostasis in whole blood. When increasing amounts of rFVIIa were added in vitro to whole blood samples, significant decreases in the values of the clotting time and clot formation time and a significant increase in the maximum clot firmness were observed. Cord whole blood was significantly more sensitive to rFVIIa addition than adult whole blood, an effect probably attributable to the low anticoagulant capacity of the neonatal plasma. Maximum clot firmness values were significantly lower in cord whole blood than in adult whole blood, an effect mainly attributable to the hypofunctional state of neonatal platelets. Since cord whole blood exerted a significantly higher sensitivity to addition of rFVIIa, we speculate that lower doses of rFVIIa might be required to treat neonates with bleeding tendency compared with the adult rFVIIa administration strategy. PMID- 16269938 TI - Venous and arterial thrombosis following administration of intravenous immunoglobulins: reply to Feuillet and colleagues. PMID- 16269939 TI - Food for thought on family dinners and holistic health. PMID- 16269941 TI - The healing effects of crying. AB - How does crying improve and enhance the emotional state? A personal, life changing event experienced by the author helped postulate an answer to this question. This article explores tear production, crying across cultures and eras, the gender factor in tears and crying, and the author's ethnographic study of Pakistani men and women's views of crying in terms of its spiritual, cultural, and psychosomatic ramifications. Greater understanding of natural human emotions and the language of tears benefits the nursing profession and gives a better understanding of the phenomenon of crying. PMID- 16269943 TI - The therapeutic use of music on agitated behavior in those with dementia. AB - Clinically, nursing has long recognized dementia behaviors--agitation, aggression, wandering, and general confusion--to be a significant patient problem as well as a major stress to nursing staff and caregivers. Although there is no cure, much can be done to improve these patients' daily care and, ultimately, their quality of life. PMID- 16269945 TI - The importance of words: suggesting comfort rather than pain. AB - Human beings shape their words; however, human beings are also shaped by their words. Although the nursing profession has paid significant attention to recognition and treatment of pain, this article illustrates how a nurse's preoccupation with pain or even the exaggerated use of the word itself can, paradoxically, evoke or aggravate the phenomenon of pain. Several vignettes, including the author's personal story, provide anecdotal evidence. PMID- 16269944 TI - The Sh-h-h-h Project: nonpharmacological interventions. AB - Promoting rest and sleep is integral to the profession of nursing. The Sh-h-h-h Project, a nonpharmacological program designed to enhance rest and sleep, was implemented on a hospital medical unit. Nursing assistants provided patients with various modalities to improve sleep, including back rubs, warm drinks, blankets warmed in a blanket warmer, aromatherapy, relaxation music, and earplugs. Additional interventions were taken to reduce noise. The outcomes of the Sh-h-h-h Project are reported here, with patients indicating improved sleep quality and quantity. PMID- 16269946 TI - Story theory: advancing nursing practice scholarship. AB - Stories are a fundamental dimension of human experience and nursing practice. Story theory describes a narrative happening that occurs through intentional nurse-person dialogue. Seven inquiry phases are associated with story theory, including gathering the story, reconstructing the story, connecting it to the literature, naming the complicating health challenge, describing the story plot, identifying movement toward resolving, and gathering additional stories. This article describes the use of story theory to advance nursing practice scholarship in both academic and everyday nursing practice. PMID- 16269947 TI - Enhancing women's mood and energy: a research-based program for subthreshold depression using light, exercise, and vitamins. AB - The prevalence and clinical significance of subthreshold forms of depression with sequelae comparable to major depression have been recently described in the literature; however, research on effective treatment is rare. A new intervention program that combines a specific regimen of light, exercise, and vitamins is effective in improving women's mood and overall sense of well-being. This program is well suited to many patients who present with somatic and psychological symptoms consistent with subthreshold depression. PMID- 16269948 TI - Spiritual distress in adult cancer patients: toward conceptual clarity. AB - Spiritual distress is conceptualized as impairments in 7 constructs of a person's sense of spirituality: (1) connectedness, (2) faith and religious belief system, (3) value system, (4) meaning and purpose in life, (5) self-transcendence, (6) inner peace and harmony, and (7) inner strength and energy. This article clarifies spiritual distress through concept analysis and provides nurses with cues for its recognition in adult patients with cancer. PMID- 16269949 TI - Get familiar with glucosamine. PMID- 16269950 TI - Angiotensin II receptor blockers and myocardial infarction: deeds and misdeeds. AB - BACKGROUND: A recent editorial published by Verma and Strauss, entitled 'Angiotensin receptor blockers and myocardial infarction', examined, through a partial analysis of individual trials, the use of angiotensin receptor blockers (ARBs) in a variety of clinical settings. This editorial was reported widely in the lay press and media, and generated disappointment and concern among physicians in many countries, probably because of its provocative subtitle in the British Medical Journal: 'These drugs may increase myocardial infarction and patients may need to be told'. OBJECTIVE AND METHODS: In order to explore the influence of ARBs on myocardial infarction, we performed a more comprehensive and updated meta-analysis, taking into account all major international, randomized trials using ARBs compared with another active drug or conventional therapy (placebo), and reporting information on rates of myocardial infarction. RESULTS: We found no significant differences in fatal and non-fatal myocardial infarction between treatment with ARBs, placebo or active treatment, and the same result was obtained when considering only trials in which ARBs were compared with angiotensin-converting enzyme inhibitors (ACEIs), or when pooling all trials together. The pooled analysis of these trials shows that the relative risk of myocardial infarction lies substantially on the indifference line. CONCLUSION: Our analysis demonstrates that, at this time, there is no evidence of increased risk of myocardial infarction in patients treated with ARBs. PMID- 16269951 TI - Do we need to target 'prediabetic' hypertensive patients? AB - Patients at risk for diabetes development have been recently characterized as those presenting higher baseline serum glucose concentration, increased body mass index, elevated systolic blood pressure, reduced serum high-density lipoprotein cholesterol and those with history of prior use of antihypertensive drugs. Little is known, however, about the long-term outcome of patients at high risk for diabetes development, so-called 'prediabetic' patients. Prediabetes state has been defined as the presence of either impaired glucose tolerance or impaired fasting glucose, and accumulating evidence suggests that individuals with a non diabetic range of hyperglycaemia (prediabetic) are already at risk for cardiovascular diseases. This short review analyses the need of targeting 'prediabetic' hypertensive patients in order to develop strategies for cardiovascular protection intended to diminish the consequences of precipitating the development of diabetes and its cardiovascular and renal deleterious effects. PMID- 16269952 TI - Genetic determinants of blood pressure regulation. AB - Hypertension is a multifactorial disorder that probably results from the inheritance of a number of susceptibility genes and involves multiple environmental determinants. Existing evidence suggests that the genetic contribution to blood pressure variation is about 30-50%. Although a number of candidate genes have been studied in different ethnic populations, results from genetic analysis are still inconsistent and specific causes of hypertension remain unclear. Furthermore, the abundance of data in the literature makes it difficult to piece together the puzzle of hypertension and to define candidate genes involved in the dynamic of blood pressure regulation. In this review, we attempt to highlight the genetic basis of hypertension pathogenesis, focusing on the most important existing genetic variations of candidate genes and their potential role in the development of this disease. Our objective is to review current knowledge and discuss limitations to clinical applications of genotypic information in the diagnosis, evaluation and treatment of hypertension. Finally, some principles of pharmacogenomics are presented here along with future perspectives of hypertension. PMID- 16269953 TI - More heat to support aggressive blood pressure lowering: the FEVER study. PMID- 16269954 TI - Estrogen-related genes and blood pressure: an example of the complexity of advanced association studies. PMID- 16269955 TI - Cardiotrophin: its importance as a pathogenetic factor and as a measure of left ventricular hypertrophy. PMID- 16269956 TI - Does the metabolic syndrome make blood pressure control more difficult in diabetes? PMID- 16269957 TI - The Felodipine Event Reduction (FEVER) Study: a randomized long-term placebo controlled trial in Chinese hypertensive patients. AB - OBJECTIVE: To compare the incidence of stroke and other cardiovascular events in hypertensive patients receiving a low-dose diuretic and low-dose calcium antagonist combination with those receiving low-dose diuretic monotherapy, and assess the effects of a small blood pressure difference at achieved levels lower than those achieved in previous placebo-controlled trials. METHODS: The Felodipine Event Reduction (FEVER) trial was an investigator-designed, prospective, multicentre, double-blind, randomized, placebo-controlled, parallel group trial. It enrolled 9800 Chinese patients, of either sex, aged 50-79 years, with one or two additional cardiovascular risk factors or disease, whose blood pressure, 6 weeks after switching from previous antihypertensive therapy to low dose (12.5 mg a day) hydrochlorothiazide, was in the range 140-180 mmHg (systolic) or 90-100 mmHg (diastolic). These patients were randomly assigned either to low-dose felodipine extended release or placebo, and followed at 3 month intervals for an average of 40 months. RESULTS: The intention-to-treat analysis included 9711 randomly selected patients with only 30 (0.3%) lost to follow-up. A total of 31 842 patient-years of follow-up were accumulated, with 85.9% of patients remaining on blinded randomized treatment. Add-on therapy was given to 33.9% of the hydrochlorothiazide-felodipine patients and to 42.3% of the hydrochlorothiazide-placebo patients. In the felodipine group, systolic blood pressure (SBP)/diastolic blood pressure (DBP) decreased (from randomization to study end) from 154.2/91.0 to 137.3/82.5 mmHg, and in the placebo group from 154.4/91.3 to 142.5/85.0 mmHg, with an average difference throughout the trial of 4.2/2.1 mmHg. In the felodipine group, the primary endpoint (fatal and non-fatal stroke) was reduced by 27% (P = 0.001). Among secondary endpoints, all cardiovascular events were reduced by 27% (P < 0.001), all cardiac events by 35% (P = 0.012), death by any cause by 31% (P = 0.006), coronary events by 32% (P = 0.024), heart failure by 30% (P = 0.239), cardiovascular death by 33% (P = 0.019), cancer by 36% (P = 0.017) in the felodipine group. No significant differences were found in new-onset diabetes. Both treatments were very well tolerated. CONCLUSIONS: In moderately complicated hypertensive patients from China even a difference in SBP/DBP as small as 4/2 mmHg, such as that induced by adding low-dose felodipine to low-dose hydrochlorothiazide, is associated with very substantial reductions in the incidence of most types of cardiovascular events. As the SBP achieved in the felodipine group was below the recommended goal of less than 140 mmHg, and SBP in the placebo group was slightly above that level, FEVER provides the required evidence in support of the guidelines recommended goal, even for a hypertensive population not entirely consisting of patients with diabetes or previous cardiovascular events. PMID- 16269958 TI - A longitudinal study on the relationship between shift work and the progression of hypertension in male Japanese workers. AB - OBJECTIVE: In this 1991-2001 cohort study of 6495 male workers in a Japanese steel company, we investigated whether shift work affects progression from mild hypertension to severe hypertension. DESIGN: A prospective cohort study. PARTICIPANTS: Participants had mild hypertension at entry into the study with systolic blood pressure of 140-159 mmHg or diastolic blood pressure of 90-99 mmHg or both before treatment. In summary, 2911-2941 workers were included for each endpoint. Pooled logistic regression analysis was performed to adjust for the effect of contributing factors and annual variations. MAIN OUTCOME: Either severe hypertension (systolic blood pressure >or= 160 mmHg and/or diastolic blood pressure >or= 100 mmHg), severe systolic hypertension (systolic blood pressure >or= 160 mmHg) or severe diastolic hypertension (diastolic blood pressure >or= 100 mmHg) was defined as a different endpoint of observation. RESULTS: Job schedule was significantly associated with progression from mild hypertension to severe hypertension and severe diastolic hypertension. The odds ratios of shift workers compared with regular day workers were 1.23 (95% confidence interval: 1.05-1.44) and 1.28 (95% confidence interval: 1.07-1.52) for severe hypertension and severe diastolic hypertension, respectively. Age, body mass index, creatinine, gamma-glutamyl transpeptidase, and glycosylated hemoglobin A1c were also significantly associated with severe hypertension, severe systolic hypertension and severe diastolic hypertension. CONCLUSION: Our study revealed that shift work is a significant and independent risk factor for the progression of hypertension. PMID- 16269959 TI - Genetic determinants of metabolic syndrome components in the stroke-prone spontaneously hypertensive rat. AB - OBJECTIVE: The metabolic syndrome is a complex multifactorial disease, which results from interactions between genes on multiple chromosomes and environmental factors. Animal models may facilitate genetic analysis of complex phenotypes by allowing complete control of environmental conditions and the ability to produce designer strains. METHODS: Stroke-prone spontaneously hypertensive (SHRSP) and Wistar-Kyoto (WKY) rat strains were used to construct congenic (SP.WKYGla2a), consomic (SP.WKYGlaYw, WKY.SPGlaYs) and double-introgressed (SP.WKYGla2aYw) strains, which were characterized for metabolic syndrome phenotypes (systolic blood pressure, glucose tolerance and lipid profile) after feeding a 60% fructose diet for 14 days. RESULTS: The Y consomic strain (SP.WKYGlaYw) demonstrated that the WKY Y chromosome significantly lowered triglyceride levels (3.77 +/- 0.60 versus 9.09 +/- 1.47 mmol/l; P < 0.001) and improved glucose tolerance [area under the curve (AUC): 26.93 +/- 0.81 versus 31.47 +/- 0.89; P < 0.05] compared with SHRSP. The chromosome 2 congenic strain (SP.WKYGla2a) exhibited significantly improved glucose tolerance (AUC: 28.19 +/- 1.17 versus 31.47 +/- 0.89; P < 0.05) and lower systolic blood pressure (161.2 +/- 6.2 versus 179.7 +/- 3.9 mmHg; P < 0.05) compared with SHRSP. 2 x 2 factorial ANOVA identified a significant interaction for glucose metabolism (P = 0.004) in the double introgressed strain (SP.WKYGla2aYw) between chromosome 2 and Y. CONCLUSIONS: These results identify novel interacting regions on chromosome 2 and the Y chromosome influencing a cluster of metabolic and cardiovascular phenotypes. Translation to clinical studies will facilitate genetic dissection of human metabolic syndrome. PMID- 16269960 TI - Impact of genes related to immune tolerance and inflammation (tumour necrosis factor-alpha, interleukin-6) on blood pressure, protein excretion and oedema in pregnancy. AB - OBJECTIVE: To test the hypothesis that genetically determined alterations of maternal immune tolerance to a foetal semi-allograft are important for the pathogenesis of hypertensive disorders in pregnancy. DESIGN: A genetic association study was performed to analyse the impact of genetic polymorphisms known to be involved in immune tolerance on markers of pre-eclampsia. SETTING: The study was conducted at the Obstetrics Department of the Charite University Hospital, Berlin, Germany. PARTICIPANTS: A total of 1480 Caucasian women were consecutively included after delivery and genotyped for two polymorphisms: tumour necrosis factor-alpha -308G>A and interleukin-6 -174G>C. MAIN OUTCOME MEASURES: Systolic and diastolic blood pressures, urinary protein excretion and oedema during pregnancy. RESULTS: Only women carrying at least one mutant allele of both polymorphisms (tumour necrosis factor-alpha A and interleukin-6 C) have a significantly elevated mean systolic blood pressure and diastolic blood pressure at the end of pregnancy. The tumour necrosis factor-alpha A allele on its own is significantly associated with urinary protein excretion in the last trimenon, and the interleukin-6 C allele is independently and significantly associated with new onset oedema. CONCLUSIONS: We demonstrate in a large population that common maternal polymorphisms of genes related to immune tolerance and inflammation are associated with blood pressure regulation, urinary protein excretion and oedema during pregnancy. The analysed polymorphisms seem to contribute to the multifactorial pathogenesis of gestational hypertension and pre-eclampsia. The findings support the hypothesis that genetically determined factors of maternal immune tolerance play a role in the pathogenesis of hypertensive disorders in pregnancy. PMID- 16269961 TI - Variation in estrogen-related genes and cross-sectional and longitudinal blood pressure in the Framingham Heart Study. AB - OBJECTIVE: To examine the association between variation in estrogen-related genes and cross-sectional and longitudinal blood pressure in men and women. DESIGN: In 1780 unrelated members of the community-based Framingham Heart Study offspring cohort, systolic blood pressure and diastolic blood pressure were measured over a total of six examination cycles encompassing 24 years of follow-up. Multivariate regression analyses were used to assess the relation between untreated cross sectional and longitudinal blood pressure and polymorphisms at the estrogen receptor-alpha (ESR1), estrogen receptor-beta (ESR2), aromatase (CYP19A1), and nuclear receptor coactivator 1 (NCOA1) genes after adjustment for common risk factors. RESULTS: In men, systolic blood pressure and pulse pressure (systolic blood pressure minus diastolic blood pressure) were associated with two polymorphisms in ESR1, while pulse pressure was also associated with variations in NCOA1 and CYP19A1. Polymorphisms in ESR1, CYP19A1, and NCOA1 were associated with diastolic blood pressure in women. CONCLUSIONS: Although the underlying relations between genes involved in estrogen action and hypertension remain to be completely understood, our findings provide suggestive evidence of gender specific contributions of estrogen-related genes to blood pressure variation. As no correction for multiple testing was performed in the analyses, we view these results as suggestive and not definitive. Further studies are warranted to confirm these results using a comprehensive set of polymorphisms in order to shed more light on the involvement of estrogen in blood pressure regulation. PMID- 16269962 TI - Cardiovascular and metabolic phenotypes in relation to the ADRA2B insertion/deletion polymorphism in a Chinese population. AB - OBJECTIVE: The functional ADRA2B I/D polymorphism is associated with various cardiovascular and metabolic phenotypes in Caucasians. The purpose of our study was to investigate whether the ADRA2B I/D polymorphism is associated with such phenotypes in a Chinese population. METHODS: We enrolled 247 women and 234 men in a family-based Chinese study. Our statistical methods included generalized estimating equations and quantitative transmission disequilibrium test. RESULTS: The I allele (62.3 versus 50.8%, P = 0.015) and the II genotype (40.9 versus 23.4%, P = 0.017) were more prevalent among hypertensive than normotensive men. While adjusting for covariates and family clusters, male II homozygotes compared with D-allele carriers had higher systolic pressure (130.0 versus 125.0 mmHg, P = 0.016) and a 2.61 times greater (P = 0.008) risk of hypertension. On the other hand, II homozygous men had lower body weight (65.4 versus 69.6 kg, P = 0.008), body mass index (23.4 versus 24.5 kg/m2, P = 0.037), waist-to-hip ratio (0.838 versus 0.857, P = 0.024), serum insulin concentration (9.5 versus 13.2 mU/l, P = 0.026) and insulin resistance (homeostasis model assessment index 2.4 versus 3.2, P = 0.051). None of these associations reached statistical significance in women. In 65 informative male offspring, transmission of the I allele was associated with higher systolic pressure (+ 6.0 mmHg, P = 0.10), diastolic pressure (+ 5.5 mmHg, P = 0.021), and faster pulse rate (+ 5.8 beats/min, P = 0.019). CONCLUSION: In Chinese men, the I allele of the ADRA2B gene is associated with higher blood pressure, but also with a more favourable metabolic phenotype. PMID- 16269963 TI - Reversal of fructose-induced hypertension and insulin resistance by chronic losartan treatment is independent of AT2 receptor activation in rats. AB - OBJECTIVES: To examine whether angiotensin II type 2 receptors (AT2R) are involved in the reversal of fructose-induced hypertension and insulin resistance after chronic angiotensin II type 1 receptor (AT1R) blockade. METHODS: Sprague Dawley rats on fructose-enriched or regular diets were pretreated with losartan, an AT1R antagonist, or vehicle for 2 weeks before two-step glucose and insulin clamp experiments with [3-3H]glucose infusion. The hepatic glucose production (HGP) and whole-body glucose uptake (WBGU) were calculated during basal, euglycemic and euglycemic hyperinsulinemic periods. Blood pressure was measured before and after acute losartan (10 mg/kg, i.v. bolus), alone or in combination of PD123319 (PD, 50 microg/kg per min), an AT2R antagonist, or CGP42112 (2 microg/kg per min), an AT2R agonist, during the clamp study. RESULTS: In rats on a regular diet, acute infusion of losartan alone or in combination with PD, an AT2R blocker, did not alter blood pressure and glucose metabolism during experiments. Fructose feeding for 6 weeks significantly increased blood pressure and attenuated insulin-mediated suppression of HGP and stimulation of WBGU. Both acute and chronic administration of losartan suppressed fructose-induced hypertension. Concomitant treatment with PD and losartan blunted the acute but not chronic losartan-mediated depressor effect. Acute losartan treatment further reduced insulin-induced suppression of HGP, but simultaneously increased insulin stimulated WBGU. These acute metabolic effects of losartan were eliminated when PD was co-administered with losartan. Conversely, chronic losartan pretreatment significantly enhanced suppression of HGP and increased stimulation of WBGU by insulin, which were not altered when PD or CGP 42112 was superimposed on losartan during the clamp experiments. CONCLUSIONS: These results suggest that reversal of high fructose-induced hypertension and insulin resistance by chronic losartan treatment is not dependent on AT2R activation and that functional activation of AT1R plays a major role in the pathogenesis of high fructose-induced hypertension and insulin resistance. PMID- 16269964 TI - Blockade of angiotensin II provides additional benefits in hypertension- and ageing-related cardiac and vascular dysfunctions beyond its blood pressure lowering effects. AB - OBJECTIVES: To assess the blockade of the renin-angiotensin system (RAS) or blood pressure-lowering on cardiovascular functions in hypertensive and ageing animals. METHODS: Male spontaneously hypertensive rats (SHR) and their normotensive counterparts, Wistar-Kyoto rats (WKY), at the ages of 3-4 (young), 34-35 (adult) and 74-75 (old) weeks were treated with an angiotensin II type 1 receptor antagonist, losartan (25 mg/kg) or a combination of a smooth muscle relaxant and a diuretic [H/H, hydralazine (50 mg/kg) plus hydrochlorothiazide (7.5 mg/kg), respectively] for 8 weeks. Each experimental group contained 10 SHR and 10 WKY, where equal numbers of untreated animals served as controls. RESULTS: Compared to age-matched WKY groups, SHR groups possessed, on average, 48 +/- 7 mmHg and 57 +/ 16 mmHg (P < 0.05) higher systolic blood pressure and left ventricular developed pressures, respectively. The values of these parameters were significantly lowered in both strains by both treatment regimens. SHR had higher heart rates, which were increased by H/H treatment selectively in adult and old animal groups of both strains. Both treatment regimens enhanced KCl-mediated, that is, receptor independent, aortic contractile responses and bradykinin-mediated coronary vasodilatation in adult and old WKY and SHR age-groups. Although both therapies augmented endothelium-dependent and endothelium-independent relaxant responses in young and adult, but not in old, SHR aortas to the levels observed in age-matched WKY, these beneficial effects were more prominent with losartan. Moreover, losartan reduced heart to body weight ratio in all SHR age groups, and selectively in the old WKY group. CONCLUSIONS: Blockade of RAS provides a better protective effect on cardiovascular function compared to sole reduction of blood pressure, and the efficacy of antihypertensive treatment is dictated by age and the hypertensive stage of the animals. PMID- 16269965 TI - Castration reduces blood pressure and autonomic venous tone in male spontaneously hypertensive rats. AB - OBJECTIVE: The development of arterial hypertension is sexually dimorphic. Venous tone is elevated in the spontaneously hypertensive rat model of hypertension. This study tested the hypothesis that endogenous androgens exacerbate venous tone in the developmental stages of spontaneous hypertension. METHODS: Male spontaneously hypertensive rats (SHRs) were subjected to sham operation, castration or castration + testosterone treatment. Ten-week-old SHR rats were instrumented for the measurement of arterial and venous pressure. A balloon catheter was advanced into the right atrium. Mean circulatory filling pressure (MCFP), an index of venous tone, was calculated. Mean arterial pressure (MAP) and MCFP were recorded from conscious rats. Postsynaptic adrenergic responsiveness was assessed by constructing cumulative dose-response curves to norepinephrine (NE). Baseline values and responsiveness to NE were obtained before and after autonomic blockade. RESULTS: MAP and MCFP were significantly reduced in castrated (MAP, 130 +/- 4 mmHg; MCFP, 5.5 +/- 0.2 mmHg) versus sham-operated SHRs (MAP, 149 +/- 5 mmHg; MCFP, 6.7 +/- 0.3 mmHg) or castrated + testosterone-treated SHRs (MAP, 145 +/- 6 mmHg; MCFP, 7.1 +/- 0.4 mmHg). Ganglion blockade abolished these differences in MAP and MCFP. Infusion of NE caused dose-dependent increases in MAP and MCFP. The MAP responses in castrated SHRs were displaced to the right of those for sham and castrated + testosterone-treated SHRs. This was not evident in the venous circulation, where there were no marked differences in the NE dose MCFP response curves. CONCLUSION: Accordingly we conclude that endogenous male sex steroids contribute to the elevated arterial and venous pressures observed in the SHR. PMID- 16269966 TI - Mechanisms for nongenomic and genomic effects of aldosterone on Na+/H+ exchange in vascular smooth muscle cells. AB - OBJECTIVES: We have reported that exposure of vascular smooth muscle cells (VSMCs) to aldosterone for 3 and 24 h activated Na+/H+ exchange (NHE) via nongenomic and genomic mechanisms, respectively. The present study determined whether aldosterone-induced nongenomic and genomic NHE activation depends on the number of transporters, the turnover rate of a single transporter, and/or the change in intracellular pH (pHi) sensitivity of the transporter, and whether aldosterone-induced NHE activation is inhibited by the selective mineralocorticoid receptor (MR) antagonist (eplerenone). METHODS: Using a fluorescent dye, we assessed NHE activity by Na-dependent acid extrusion rates (JH) after an acid load in the absence of CO2/HCO3- in VSMCs treated with aldosterone. RESULTS: Treatment with aldosterone for 3 and 24 h increased JH at the wide pHi range, and shifted the JH versus pHi in the alkaline direction. Without affecting the apparent Km for external Na+, the Vmax increased in VSMCs treated with aldosterone for 3 and 24 h. Both eplerenone and spironolactone inhibited only aldosterone-induced genomic NHE activation, but the IC50 of eplerenone was smaller than that of spironolactone. CONCLUSION: We demonstrated that: (1) both nongenomic and genomic stimulatory effects of aldosterone on NHE activity in VSMCs occur by an increase in the number of NHEs and the alkaline shift in pHi sensitivity of the NHE; (2) only the aldosterone-induced genomic NHE activation occurs via MR; and (3) both eplerenone and spironolactone inhibit the aldosterone-induced genomic NHE activation, but eplerenone is more effective than spironolactone, based on the IC50 value in VSMCs. PMID- 16269967 TI - Oxidative stress contributes to the enhanced expression of Gialpha proteins and adenylyl cyclase signaling in vascular smooth muscle cells from spontaneously hypertensive rats. AB - OBJECTIVE: We have previously shown an enhanced expression of Gialpha proteins in spontaneously hypertensive rats (SHR) that precedes the development of hypertension. Since oxidative stress has been shown to be increased in SHR, the present studies were undertaken to examine the role of oxidative stress in enhanced expression of Gialpha proteins in SHR. METHODS: Aortic vascular smooth muscle cells (VSMC) from 12-week-old SHR and Wistar-Kyoto (WKY) rats were used for the present studies. The levels of inhibitory guanine nucleotide regulatory proteins (Gialpha-2 and Gialpha-3) and stimulatory proteins (Gsalpha) were determined by western blotting techniques. Adenylyl cyclase activity was determined by measuring [32P]cAMP formation from [alpha-32P]ATP. RESULTS: VSMC from SHR exhibited enhanced expression of Gialpha-2 and Gialpha-3 proteins as compared with age-matched WKY rats; however, the levels of Gsalpha proteins were not different between the two groups. The levels of superoxide anion (O2-) were also increased in SHR as compared with WKY rats, and losartan, an AT1 receptor antagonist, restored the enhanced levels to control WKY rat levels. Treatment of VSMC with antioxidants such as N-acetyl-L-cysteine (NAC) or diphenyleneiodonium (DPI) for 24 h decreased the enhanced expression of Gialpha-2 and Gialpha-3 proteins in a concentration-dependent manner in VSMC from SHR. In addition, the inhibition of forskolin-stimulated enzyme activity by low concentrations of GTPgammaS (receptor-independent Gi functions) and C-ANP4-23-mediated inhibition of adenylyl cyclase (receptor-dependent Gi functions) that were significantly enhanced in SHR were restored to WKY rat levels by NAC and DPI treatments. Similarly, diminished stimulation of adenylyl cyclase by GTPgammaS, isoproterenol and sodium fluoride in SHR was also restored towards control WKY rat levels by NAC and DPI treatments. Furthermore, PD98059, a selective inhibitor of mitogen activated protein kinase, was able to restore the enhanced expression of Gialpha proteins in VSMC from SHR towards WKY rat levels. In addition, the enhanced activity of extracellular signal-regulated kinase 1/2 in SHR as compared with WKY rats, as demonstrated by enhanced phosphorylation of extracellular signal regulated kinase 1/2, was also restored to WKY rat levels by NAC or DPI. CONCLUSIONS: These results suggest that enhanced levels of Gialpha proteins and associated functions in SHR may be attributed to the enhanced oxidative stress present in SHR, which exerts its effects through the mitogen-activated protein kinase signaling pathway. PMID- 16269968 TI - Serum tissue inhibitors of metalloproteinases 1 (TIMP-1) and carotid atherosclerosis and aortic arterial stiffness. AB - BACKGROUND: The results of experimental studies have suggested that matrix metalloproteinases (MMPs) and their inhibitors, the tissue inhibitors of metalloproteinases (TIMPs), are involved in vascular remodeling. In a population based study, we report the relationships of serum TIMP-1 with carotid intima media thickness, carotid plaques and aortic arterial stiffness. METHODS: Free health examinations were performed on 238 men free of coronary heart diseases (aged 56.5 +/- 10.4 years, 57.1% were hypertensive). Carotid-femoral pulse-wave velocity (PWV) was used to assess aortic stiffness. Carotid ultrasound examination included measurements (at sites free of plaques) of intima-media thickness (IMT) at the common carotid arteries (CCA) and assessment of atherosclerotic plaques in the extracranial carotid arteries. RESULTS: The percentage of subjects with plaques was lower in subjects with low TIMP-1 values (P for trend = 0.0001). In multivariate analysis adjusted for age, body mass index, smoking habits, total cholesterol, triglycerides, C-reactive protein, heart rate, diabetes and systolic blood pressure, the odds ratio of carotid plaques in subjects with high values of TIMP-1 (tertile 3) compared to those with low values (tertile 1) was 2.89 (95% confidence interval 1.12-7.47, P < 0.01). TIMP-1 was positively associated with CCA-IMT and PWV in univariate analysis. However, the associations disappeared once age and systolic blood pressure were taken into account in the multivariate analysis. CONCLUSION: This study shows that there is a differential association of TIMP-1 with PWV, CCA-IMT and carotid plaques. Our results suggest that TIMP-1 might be involved in plaque formation. PMID- 16269969 TI - Menopause-related blood pressure increase and its relationship to age and body mass index: the SIMONA epidemiological study. AB - BACKGROUND: Menopause is commonly associated with some blood pressure (BP) rise, but cross-sectional or longitudinal studies completed so far were often too small and were unable to indicate whether this BP increase is really dependent on menopause, or was caused by age or changes in body mass index (BMI). METHODS AND RESULTS: The SIMONA study (Study on Hypertension Prevalence in Menopause in the Italian population) was a large cross-sectional study on 18 326 women of age range 46-59 years, consecutively seen by 302 practitioners all over Italy, and representing 60% of the women of that age in the National Health care list of those doctors. BP was measured three times in the seated position by the same automatic machine, and demographic and clinical data were taken. Systolic blood pressure (SBP) and diastolic blood pressure (DBP) were slightly but significantly higher in postmenopausal than premenopausal and perimenopausal women, but so were age and BMI. Within seven biannual strata, differences in age and BMI were minimized, but SBP/DBP remained significantly higher (by 3.4/3.1 mmHg) in postmenopausal than in premenopausal subjects in the youngest stratum (46-47 years), and was also significantly higher in the stratum 48-49 years. The differences remained significant after the exclusion of 1809 women with surgical menopause or 695 women with cardiovascular disease. Even when the confounding effects of age, BMI, smoking and contraceptive or replacement therapies were excluded by analysis of covariance, menopause was significantly and positively associated with SBP and DBP (approximately 2 mmHg difference in the age range 46 49 years). CONCLUSION: Menopause is associated with a slightly but significantly higher BP, even after adjustment for age and BMI, as well as other confounding factors, but the association is evident only in the younger end of the age range related to menopause. PMID- 16269970 TI - Low-dose lithium combined with captopril prevents stroke and improves survival in salt-loaded, stroke-prone spontaneously hypertensive rats. AB - OBJECTIVE: A number of potential interactions between angiotensin-converting enzyme inhibitors and lithium have been described in the literature. In the present study, we investigated the effects of a low-dose combination treatment with lithium and captopril on survival and stroke prevention in salt-loaded, stroke-prone spontaneously hypertensive rats (SHRSP). METHODS: Eight-week-old saline-drinking SHRSP (n = 21 per group) were treated with vehicle, LiCl (1 mmol/kg per day), captopril (25 mg/kg per day) and captopril plus LiCl for up to 37 weeks. Body weight, salt water intake blood pressure and mortality were recorded throughout the experimental period. Plasma renin activity, plasma lithium concentration and urinary excretion of albumin, sodium and potassium were measured at different time points. RESULTS: Captopril treatment doubled the life expectancy when compared with vehicle-treated rats. Lithium alone had minor effects on survival but led to a dramatic increase in survival when added to captopril (mean survival time > 237 versus 147 days, P < 0.001). Systolic blood pressure increased with age in all treatment groups but was comparable in the captopril-treated and the captopril-plus-lithium-treated groups. Plasma renin activity as well as urinary sodium and potassium excretion did not differ between both groups. In the captopril group a striking fivefold increase of albuminuria occurred between 14 and 26 weeks of age, while this progression was completely abolished by the addition of lithium. CONCLUSIONS: Our results demonstrate that the addition of lithium to captopril dramatically prolong the effects of the angiotensin-converting enzyme inhibitor on survival in salt-loaded SHRSP. This effect was independent of a reduction in blood pressure. PMID- 16269971 TI - Renoprotective effect of long-term combined treatment with adrenomedullin and omapatrilat in hypertensive rats. AB - BACKGROUND: Previous studies demonstrated that adrenomedullin (AM) is metabolized by neutral endopeptidases and that the renal effect of AM is augmented by the inhibition of neutral endopeptidases. We have recently shown that the long-term administration of AM has renoprotective effects. OBJECT: This study assessed the chronic renoprotective effects of AM combined with a vasopeptidase inhibitor in hypertensive rats and attempted to elucidate the mechanism involved. METHODS: We studied the following four groups: control Dahl salt-resistant (DR) rats, untreated Dahl salt-sensitive (DS) rats, omapatrilat (35 mg/kg per day)-treated DS rats; and human AM (500 ng/h) plus omapatrilat-treated DS rats. After 7 weeks' treatment, blood pressure, renal function, neurohumoral factors, gene expression levels, and histological findings were examined. RESULTS: DS rats were characterized by increased blood pressure, decreased renal function, abnormal histological findings, and increased gene expression of collagen I and III, transforming growth factor beta (TGF-beta), and NADPH oxidase subunits (p40phox, p47phox, and gp91phox) in the renal cortex compared with DR rats. Compared with DS rats, omapatrilat significantly decreased systolic blood pressure (-26 mmHg), improved renal function, histological findings, and messenger RNA expression levels of collagen I, collagen III, and TGF-beta. Combined treatment with omapatrilat and AM further improved renal function, histological findings, and mRNA expression levels of collagen I, collagen III, and TGF-beta, without a further reduction in blood pressure. Only combined treatment decreased mRNA levels of p40phox, p47phox, and gp91phox. There were no differences in plasma AM or atrial natriuretic peptide levels among three DS groups. CONCLUSION: Our results suggest that combined treatment with omapatrilat and AM provides additional renoprotective effects independent of blood pressure-lowering activity partly via inhibition of gene expressions of oxidative stress and extracellular matrix. PMID- 16269972 TI - Usefulness of plasma cardiotrophin-1 in assessment of left ventricular hypertrophy regression in hypertensive patients. AB - OBJECTIVE: We investigated whether regression of left ventricular hypertrophy (LVH) in response to antihypertensive treatment is associated with plasma cardiotrophin-1 (CT-1) in hypertensive patients. METHODS: The study was performed in 47 patients with mild to moderate essential hypertension, and LVH was assessed echocardiographically. The family doctor gave antihypertensive treatment and followed all patients. LVH regression was diagnosed if the baseline left ventricular mass index (LVMI) decreased to normal values after 1 year of treatment. Plasma CT-1 was determined by an enzyme-linked immunosorbent assay. RESULTS: The LVMI was normalized in 23 patients (49%) and persisted at an abnormally increased level in 24 patients (51%) after 1 year of treatment, whereas the reduction in clinic and home blood pressure was similar in the two groups: CT-1 decreased (-48%, P < 0.005) and increased (+35%, P < 0.05) in patients in whom LVH regressed and LVH persisted, respectively. Final values of CT-1 were inversely correlated (r = 0.534, P < 0.001) with the decrease in LVMI after treatment in all patients. A significant association (chi2 = 16.87, P < 0.001) was found between normalization of CT-1 and regression of LVH with treatment. A cut-off value of 41 fmol/ml for CT-1 provided a relative risk of 43.13 (95% confidence interval, 4.88-380.48) for detecting LVH regression. CONCLUSION: These results show an association between treatment-induced decrease of plasma CT-1 and LVH regression in essential hypertension. Although preliminary, these findings suggest that the determination of plasma CT-1 may be useful for the follow-up of hypertensive heart disease in routine clinical practice. PMID- 16269973 TI - Predictors of successful long-term blood pressure control in type 2 diabetic patients: data from the Swedish National Diabetes Register (NDR). AB - BACKGROUND: Hypertension in patients with diabetes is a well recognized cardiovascular risk factor for which the benefits of treatment are strongly evidence based. Less is known about predictors for successful long-term blood pressure control in these patients, including the potential role of body mass index (BMI), glycaemic control, microalbuminuria and smoking. MATERIAL AND METHODS: We used longitudinal data on risk factor levels from repeated clinical surveys of 1759 type 2 diabetic patients in the Swedish National Diabetes Register (NDR), a nationwide annual registration of quality indicators in diabetes care. Subjects with successful blood pressure (BP) control (systolic BP < 135 mmHg and diastolic BP < 85 mmHg) at baseline in 1997, in 2001, and at follow-up in 2003, were compared to subjects with BP control >or= 135/85 mmHg. RESULTS: Logistic regression analysis disclosed that successful BP control during the study period was predicted by lower BMI (P < 0.001), a lower frequency of microalbuminuria (P = 0.002), and lower age (P < 0.001) at baseline in 1997, and was still associated with lower BMI (P < 0.001), a lower frequency of microalbuminuria (P = 0.01) and lower age (P < 0.001) at follow-up. Successful BP control was also associated at follow-up with a lower frequency of the metabolic syndrome (30 versus 75%) and lower predicted 10-year risks [United Kingdom Prospective Diabetes Study (UKPDS) Risk Engine] of coronary heart disease (14 versus 29%) and stroke (10 versus 22%) (all P < 0.001). CONCLUSION: A lower BMI and absence of microalbuminuria were strong independent predictors of long-term successful BP control in type 2 diabetic patients, also characterized by a lower frequency of the metabolic syndrome and lower 10-year risk of cardiovascular disease. This implies the long-term benefits on BP control of lifestyle measures as well as control of microalbuminuria. PMID- 16269974 TI - Hemodynamic and metabolic responses to valsartan and atenolol in obese hypertensive patients. AB - OBJECTIVE: None of the current hypertension guidelines provides specific guidance regarding pharmacological management of obese hypertensive patients. Treatment recommendations for lean hypertensives may not be simply extrapolated to obese hypertensive persons. DESIGN: Randomized, double-blind, parallel-group study with a 13-week treatment period. SETTING: Multicenter study in Germany. PATIENTS: Obese patients with mild to moderate uncomplicated essential hypertension. INTERVENTION: Patients were treated with valsartan at a maximal dose of 160 mg/day or with atenolol at a maximal dose of 100 mg/day. Hydrochlorothiazide at doses of 12.5-25 mg was added in patients with blood pressure > 140/90 mmHg on monotherapy. MAIN OUTCOME MEASURES: Blood pressure, lipid and glucose metabolism, and highly sensitive C-reactive protein (hsCRP) were monitored. RESULTS: Sixty seven patients were randomized to valsartan and 65 patients to atenolol. With valsartan, systolic blood pressure (SBP) decreased from 160.8 +/- 8.9 to 140.5 +/ 13.3 mmHg and diastolic blood pressure (DBP) from 96.1 +/- 7.0 to 85.1 +/- 8.1 mmHg by the end of the study. With atenolol, SBP decreased from 159.3 +/- 6.8 to 139.8 +/- 14.5 mmHg and DBP from 95.0 +/- 6.8 to 83.5 +/- 7.5 mmHg (P = 0.91 for SBP and P = 0.34 for DBP between interventions). Body weight did not change with either treatment. We did not see a significant difference in the response of lipid levels or hsCRP between interventions. To assess the cumulative effect of each intervention on glucose metabolism over the trial duration, we calculated individual areas under the curve for homeostasis model assessment for insulin resistance (HOMA-IR) over time. The resulting area under the curve was significantly smaller with valsartan compared with atenolol (P = 0.02). CONCLUSIONS: Beta-adrenoreceptor blockers and AT1-receptor blockers, particularly in combination with low-dose diuretics, effectively lower blood pressure in obese hypertensives. However, metabolic responses differ between both treatment strategies, with beneficial effects of AT1-receptor blockers. AT1-receptor blockers are a good choice in obese hypertensives, given the profoundly increased diabetes risk in this population. PMID- 16269976 TI - Total cholesterol and life expectancy in the elderly. PMID- 16269975 TI - Intravenous use of the calcium-channel blocker nicardipine as second-line treatment in severe, early-onset pre-eclamptic patients. AB - OBJECTIVE: To evaluate the efficacy of intravenous administration of nicardipine as a second-line temporizing treatment in severe, early-onset, pre-eclamptic patients. DESIGN: An open, prospective, evaluation study. SETTING: A high-care obstetric ward in a tertiary care centre. PATIENTS: Twenty-seven early-onset, pre eclamptic patients with a median gestational age of 27 weeks 1 day (range, 21 weeks 2 days-32 weeks 4 days) with treatment failure on standard intravenous antihypertensive drugs (ketanserin, dihydralazin or labetalol). INTERVENTION: Nicardipine infusion was started for temporizing management of pre-eclampsia at a dosage of 3 mg/h and was subsequently titrated according to blood pressure. Nicardipine treatment was continued for as long as the maternal and foetal conditions allowed. MAIN OUTCOME MEASURES: The endpoints of the study were defined as the percentage of patients reaching the target diastolic intra arterial blood pressure (< 100 mmHg or < 90 mmHg in Haemolysis, Elevated Liver Enzymes, Low Platelet Count syndrome patients) within 1 h after the start of treatment, and the number of days of prolongation of pregnancy under nicardipine treatment. Maternal and foetal side effects, foetal death and neonatal outcome were assessed. RESULTS: In all patients the target diastolic intra-arterial blood pressure was obtained within a median of 23 min (range, 5-60 min). Delivery was postponed for a median of 4.7 days (range, 1-26 days) using nicardipine treatment, in a maximum dosage ranging from 3 to 9 mg/h. Detailed haemodynamic parameters with corresponding nicardipine dosages were obtained in nine patients. In one-fifth of the patients, unwanted hypotensive periods were registered during treatment, manageable with dosage adaptation. Foetal well-being did not seem adversely affected. CONCLUSION: This evaluation shows that nicardipine is a potent antihypertensive drug and can be used for temporizing management in severe, early-onset pre-eclampsia when other antihypertensive drugs have failed. PMID- 16269977 TI - [Can pediatric radiology be saved?]. PMID- 16269978 TI - [Role of imaging in the evaluation of axillary nodes and metastatic disease from breast carcinoma]. AB - Disease staging of patients with breast cancer is based on the probability of metastatic disease, the reliability of complementary examinations, and therapeutic possibilities, evaluated on a cost/benefit basis. For regional disease staging, nodal status can be assessed by ultrasound, and the value of this approach can be optimized by imaging-guided biopsies. Ultrasound examination of nodes upstream of the sentinel node allows determination of the utility of this node and the indications for axillary resection. Work-up of metastatic spread is performed only after evaluation of risk factors for metastasis. Prior to therapy, and in the absence of any clinical warning signs for resectable tumors, there are no indications for imaging, which is reserved solely for locally advanced tumors. PMID- 16269979 TI - [Normal and abnormal meningeal enhancement: MRI features]. AB - The authors describe normal imaging of the meninges and meningeal spaces and MR (magnetic resonance) imaging findings in tumoral and nontumoral diseases. Dural or/and pial enhancement may be related to tumoral, infectious or granulomatous diseases. PMID- 16269980 TI - [Value of contrast-enhanced MR enterography in pediatric Crohn's disease: preliminary study]. AB - OBJECTIVE: Feasibility study of contrast enhanced MR enterography without enteroclysis as a new diagnostic tool for children with known or suspected Crohn's disease. METHODS: We prospectively included 15 children, 8-18 years old, with clinical suspicion of Crohn's disease. MR enterography without enteroclysis was performed on a 1,5 T clinical MR system. A total of 1000 ml of mannitol 5% was orally administered 60 minutes prior to MRI. Coronal and axial breath-hold sequences were acquired. The following sequences were obtained: True-FISP, FLASH T1 2D/3D with Fat saturation before and after gadolinium injection. Two radiologists, blinded to patient information, independently reviewed all examinations to record image quality, the degree of distension of the distal ileum, the presence of abnormal bowel segments and the presence of extra intestinal complications. MRI findings were correlated to sonographic, endoscopic and biological results (sensitivity, specificity, Kappa test). RESULTS: The examinations were considered of satisfactory diagnostic quality in 93.3% of patients. Respiratory artifacts were present in one case. The entire GI tract could be identified on all sequences. Distention of the distal ileum was recorded as good to excellent in 89% of healthy subjects. Five MR examinations were considered abnormal with isolated ileal involvement in 2 cases, ileocolic involvement in 2 cases, and isolated colonic involvement in 1 case. The sensitivity and specificity of MR for the positive diagnosis of Crohn's disease were 100% and 83% respectively. Three extra-intestinal complications were detected: one case of ileo-ileal fistula, not identified on ultrasonography, an asymptomatic anal fistula and a symptomatic inflammatory stricture. CONCLUSION: MR enterography without enteroclysis is a well tolerated, effective non invasive method in the evaluation of known or suspected Crohn's disease. Because of the absence of ionizing radiation, MR enterography should become the gold standard in pediatric patients. PMID- 16269981 TI - [Febrile osseous pain in children with sickle cell disease: MRI findings]. AB - PURPOSE: The aim of this study was to report the MRI findings that can suggest a vaso-occlusive crisis in cases of febrile osseous pain in children suffering from sickle cell disease. MATERIALS AND METHODS: MRI (T1 and T2 weighted sequences and T1 weighted sequence with fat saturation before and after gadolinium injection) was performed in 10 children with sickle cell disease, presenting with febrile osseous pain. The diagnosis of vaso-occlusive crisis was made after fast improvement due to symptomatic treatment and negative bacteriological result. RESULTS: MRI was abnormal in all cases. A multifocal localisation was found in 2 cases. Bone marrow abnormalities were constant. In 10 cases, high T1 and T2 signal and metaphyso-diaphysial lesions were noted. Heterogeneous medullar enhancement with "ink stain" feature was constant. Early periosteal abnormalities were noted in 8 cases with inflammatory or stratified features. Cortical thinning was found in 1 case. Soft tissue abnormalities were observed in 5 cases with inflammatory features in 4. CONCLUSION: Multifocal synchronous localisation, medullar abnormalities resulting from hemoglobin degradation, heterogeneous enhancement, early periosteal abnormalities and associated soft tissues swelling are MRI findings suggesting acute vaso-occlusive disease. PMID- 16269982 TI - [Abdominal wall abscess as the presenting sign in carcinoma of the colon: report of 4 cases]. AB - PURPOSE: We report four cases of unusual presentation of colonic cancer revealed by an abscess of the abdominal wall. MATERIALS AND METHODS: Our study included 3 men and 1 women ranging in age from 33 to 66 years presenting with abdominal wall abscess (in the left anterior abdominal wall in 2 patients and in the quadratus lumborum muscle in the other 2 patients). Our cases were listed from the departments of radiology and abdominal surgery of Sahloul hospital from 1995 through 2000. Ultrasonography (US) and computed tomography (CT) were performed in all cases; barium enema in three. RESULTS: US and CT revealed segmental colonic wall thickening (left colon in three patients and right colon in one patient). The diagnosis of colonic cancer was reached by colonoscopy with biopsy in all 4 cases. All of our patients underwent surgical treatment. CONCLUSION: In this era of newer diagnostic imaging modalities, US and CT must frequently be used in the evaluation of patients with abdominal wall abscess, to depict intra-abdominal infection and especially malignant lesion causing abdominal wall abscess. PMID- 16269983 TI - [Aortic diameter measurements on MRI follow-up of coarctation]. AB - OBJECTIVE: During aortic coarctation follow-up, accurate diameter measurements using MRI are required. The purpose of this study was to compare the reproducibility of aortic measurements on axial and sagittal views using black blood (Fast-Spin-Echo) and three-dimensional Gadolinium-enhanced MR angiography. SUBJECTS AND METHODS: We studied 34 patients who underwent evaluation after surgical treatment in the first year of life. The aortic measurements were made at six thoracic levels. We calculated the mean difference between the two sequences, and used Bland and Altman method and the comparison to zero. RESULTS: With black blood, either on axial or sagittal views, aortic measurements were in agreement. Between two sequences, we considered as acceptable a mean difference of 0,5 mm with limits of agreement of +/- 3 mm. Difficulties were found for proximal ascending aorta measurements on all sequences. Using MR angiography, the highest agreement at the site of repair was on sagittal view, while the highest agreement at other sites was on axial views. CONCLUSION: Black blood sequence provides a better reproducibility for aortic measurements than native slices of MR angiography. PMID- 16269984 TI - [Hepatosplenic tuberculosis in children]. AB - Isolated tuberculous involvement of the liver and spleen is infrequent. We report such a case in a child. The differential diagnosis of isolated multi-nodular diseases of the liver and spleen is reviewed. PMID- 16269985 TI - [Elastofibroma: imaging features]. AB - Elastofibroma dorsi is a rare benign and slow growing fibro-proliferative lesion of unknown pathogenesis. It has a characteristic location (periscapular region) and a specific imaging appearance (songraphy, CT, MRI) allowing accurate prospective diagnosis. The recognition of this benign lesion avoids unnecessary biopsy and/or surgery. We report three cases of elastofibroma dorsi illustrating the characteristic features on sonography, CT and MRI. Involvement was bilateral in two cases. PMID- 16269986 TI - [Milk of calcium in a renal cyst: a case report]. AB - Milk of calcium in a renal cyst is a rare condition. The authors report a case evaluated by ultrasonography, computed tomography (CT) and magnetic resonance imaging (MRI). A 58 year - old women presented with diffuse abdominal pain. Ultrasonography showed a left upper pole cystic lesion with echogenic component in the dependent portion causing shadowing. CT showed a cyst with layering milk of calcium. On MRI, the milk of calcium sediment had low signal intensity on T1 and T2 weighted images. PMID- 16269987 TI - [Foraminal, epidural and intravertebral migration of a calcified degenerated intervertebral disk]. AB - The authors report a case of inflammatory back pain associated with radiculopathy secondary to degenerative disk calcifications migrated within the foramen, epidural space and vertebral body. The purpose of this clinical case is to illustrate this uncommon cause of radiculopathy and avoid unnecessary invasive diagnostic procedures. PMID- 16269988 TI - [What is your diagnosis? Asymptomatic polyostotic fibrous dysplasia]. PMID- 16269990 TI - [New treatment strategies for hypertension. Which guidelines and how to apply them]. AB - The diagnosis of hypertension by blood pressure measurements taken in the physician's office has been called into question by several studies. The onset of cardiovascular events appears to correlate better with ambulatory blood pressure measurements than with those taken during consultation (either "white coat" or masked hypertension). While the US, WHO, French and European guidelines diverge as to the specific antihypertensive drug among the seven classes available should be chosen for first-line treatment, there is a consensus for specific choices as a function of the type of hypertension. In any case, most treatment trials show that more than two antihypertensive drugs are often necessary. Treatment can thus begin with two drugs. The optimal target blood pressure is defined by the US JNC7 according to whether the patient also has diabetes or a nephropathy. When hypertension is uncomplicated, the target level is 140/90 mmHg. In the case of diabetes or nephropathy, it is 130/80 mmHg. In all cases, diet and exercise changes are also necessary and it is essential that patients understand them if they are to comply with them. Diastolic blood pressure remains the most important figure for those younger than 50 years, but afterwards, systolic pressure is more relevant. Aortic pressure may be more closely associated with cardiovascular risk than the blood pressure measured at the brachial artery. The concept of comprehensive management is radically modifying our behavior : the hypertensive patient is now above all a patient at high cardiovascular risk and the treatments to consider must not be limited to antihypertensive drugs but must also include treatment of other cardiovascular risk factors (aspirin, statins, smoking cessation, etc.). PMID- 16269991 TI - [New treatment strategies for type-2 diabetes?]. AB - Earlier guidelines for type 2 diabetes (Afssaps 1999 and Anaes 2000) were based on the UK Prospective Diabetes Study, published in 1998. These guidelines recommended treatment according to HbA1c value (< 6.5%, between 6.5 and 8% and > 8%): an oral antidiabetic agent for levels > 6.5% despite diet and exercise; combined metformin + sulfonylurea) if HbA1c >8%; and insulin if the latter failed. Blood pressure goals were < 130/80 mmHg, with the antihypertensives necessary to achieve it. The LDL-cholesterol target value was < 1 g/L (for primary prevention in the case of high cardiovascular risk or for secondary prevention) or between 1.3 and 1.6 g/l (primary prevention in the absence of elevated risk). Another reading of the UK study, associated with the arrival of glitazones led to a revision of these objectives with a more aggressive treatment approach ("earlier and stronger"): screen patients for type 2 diabetes earlier, set stricter goals (HbA1c < 6%), and promptly prescribe dual therapy (metformin + sulfonylurea). Should this fail, either glitazone should be added or insulin treatment begun. For most people with type 2 diabetes, the target blood pressure remains 130/80 mmHg, regardless of the type and number of antihypertensive agents necessary. The target drops to 125/75 mmHg for patients with > 300 mg/day microalbuminuria; in these cases, treatment with agents that block the renin angiotensin system (ACE inhibitors or sartans) is recommended. The LDL cholesterol target value is 1.0 or 1.6 g/L, depending on the cardiovascular risk level. But these guidelines are applied insufficiently, especially in terms of clinical and laboratory follow-up of patients and choice of treatment. All health professionals must participate in the more effective diffusion and application of these guidelines. Patient education is fundamental. The establishment of care networks for these patients seems to us to be the best tool for meeting the objectives of these guidelines. PMID- 16269993 TI - [Primary cardiorenal prevention in patients with type-2 diabetes. The Roadmap study]. AB - The incidence of type-2 diabetes is increasing throughout the world. By 2010, 350 million people will have this disease. Microalbuminuria is present in more than one third, for some at diabetes diagnosis. Rather than a complication, it is an indication of a vascular disorder that is part of the metabolic syndrome. 25% will develop end-stage kidney failure. Several studies have identified microalbuminuria or proteinuria as an independent cardiovascular risk factor. Others have shown that antihypertensive treatments acting on the renin angiotensin system (ACE inhibitors, ARBs agents) can reduce the progression of nephropathy in people with hypertension, type 2 diabetes and microalbuminuria. The "nephroprotective" effects of these drug classes, beyond their role in blood pressure reduction, are suggested by modifications in renal structure and protein expression. But no study has so far examined their value in primary prevention in persons with type 2 diabetes without--but at risk of developing- microalbuminuria. The Roadmap study (Randomized Olmesartan And Diabetes Microalbuminuria Prevention Study) of primary prevention has as its objective measurement of the impact of ARBs (olmesartan 40 mg/d) treatment on renal outcome in 4400 patients with type 2 diabetes without microalbuminuria. Follow-up of this placebo-controlled study will last for 5 years. Conducted in 200 European centers, its results are expected for 2012. PMID- 16269994 TI - Cardiovascular risk in patients with mild renal insufficiency: implications for the use of ACE inhibitors. AB - We review the evidence linking mild renal insufficiency (MRI) with increased cardiovascular risk. MRI is associated with a number of cardiovascular risk factors, including nighttime hypertension, and increased levels of lipoprotein (a), homocysteine, asymmetric dimethyl-arginine, and inflammation and insulin resistance markers and mediators. Epidemiologic evidence associates coronary artery disease and nephrosclerosis, a frequent cause of early renal insufficiency in the elderly. In a middle-aged general population MRI was found in 8% of women and 9% of men, but was not associated with cardiovascular disease. Nonetheless, in a representative sample of middle-aged British men the risk of stroke was 60% higher for the sub-group with MRI: in people at high cardiovascular risk (mostly coronary disease), the HOPE study found a 2-fold (unadjusted) or 1.4-fold (adjusted) higher incidence of cardiovascular outcomes with MRI. The combined incidence of cardiovascular death, myocardial infarction and stroke increased with the level of serum creatinine. Several studies have examined the cardiovascular risk associated with MRI in hypertension. In HDFP, as in HOPE, cardiovascular mortality increased with serum creatinine (five-fold difference in cardiovascular mortality between the lowest and the highest creatinine strata). The risk associated with renal insufficiency was independent of other classic cardiovascular risk factors. Two trials of hypertensives with low risk (HOT and a small Italian trial) found that cardiovascular outcomes approximately doubled in subjects with MRI. Another study (MRFIT) found that it was not baseline creatinine, but its increase on follow-up that predicted future cardiovascular disease. These observational data suggest that regardless of etiology MRI is a strong predictor of cardiovascular disease and is found in 10% of populations at low cardiovascular risk and in up to 30% of those at high risk. No prospective therapeutic trials aimed at reducing the cardiovascular burden in people with MRI are available. Subgroup analyses of the HOPE study indicate that angiotensin converting enzyme (ACE) inhibition with ramipril is beneficial and does not increase the risk of such side effects as acute renal failure or hyperkalemia. Thus the frequent practice of withholding ACE inhibitors from patients with mild renal insufficiency is unwarranted, especially since MRI identifies a group at high risk that appears to benefit most from treatment. Moreover, there is evidence that ACE inhibitors improve renal outcomes in renal insufficiency. Prospective studies should test the predictive power of early renal insufficiency for cardiovascular disease and prognosis with various therapeutic options. PMID- 16269992 TI - [Cardiovascular protection and prevention in patients with cardiac conditions and diabetes]. AB - Good glycemic control can have a positive effect on the course and prognosis of coronary disease during and after myocardial infarction. It also reduces the risk of its onset. Both postprandial and fasting blood glucose must be normalized. Optimal treatment of coronary patients should include reduction of risk factors and 4 drugs: a beta-blocker at an effectively beta-blocking dose, aspirin at a daily dose between 75 and 100 mg, a statin that leads to an LDL-cholesterol level less than 1 g/L, and an angiotensin-converting enzyme inhibitor, with a demonstrated dose-effect relation. Efficacy is best when compliance is considered an essential objective. PMID- 16269995 TI - [The new antithrombotic agents]. AB - Current antithrombotic agents include anticoagulants (unfractionated and low molecular-weight heparin, and antivitamin K) and platelet aggregation inhibitors (aspirin, ticlopidine, clopidogrel). Two areas are under particular investigation: specific inhibition, direct or indirect, of factor Xa and factor IIa. Pentasaccharide, an indirect anti-Xa, has proved effective in curing deep vein thrombosis and more effective than enoxaparin for prophylactic treatment after orthopedic surgery. Administered in a single subcutaneous injection daily, it has no risk of thrombocytopenia; laboratory surveillance is based on anti-Xa activity. Hirudin and melagatran act by direct thrombin inhibition. Unlike hirudin (which requires monitoring of active coagulation time or ecarin clotting time), melagatran requires no laboratory monitoring. It is not associated with an increased risk of hemorrhage. But there is no true antidote at this time. If its efficacy is confirmed, ximelagatran, the orally active prodrug of melagatran, may facilitate the long-term treatment now reserved for antivitamin K. Three antagonists of the tissue factor-factor VIIa complex are also under development: rNAPc2 (Recombinant Nematode Anticoagulant Protein C2), ASIS (Active Site Inhibitor Factor Seven) and recombinant TFPI (Tissue Factor Pathway Inhibitor). Antiplatelet drugs are the reference antithrombotic agents for the prevention and treatment of arterial thrombosis. Aspirin remains in first place (75 to 300 mg/d) but the modest superiority of the thienopyridines (clopidogrel and ticlopidine) is established. Hemogram monitoring is no longer necessary for clopidogrel. Use of aspirin + a thienopyridine after placement of a coronary stent has been validated. Laboratory monitoring of antiplatelet treatments has not been codified. PMID- 16269996 TI - [Preventing cerebrovascular accidents during atrial fibrillation]. AB - Atrial fibrillation, the most commonly encountered arrhythmia in clinical practice, is associated with substantial morbidity and mortality. Its incidence and prevalence are increasing, and it represents a growing clinical and economic burden. Recent research has highlighted new approaches to both pharmacological and non-pharmacological management. Pooled data from trials comparing antithrombotic treatment with placebo show that warfarin reduces the risk of stroke by 62% and that aspirin alone reduces the risk by 22%. Overall, in high risk patients, warfarin was better than aspirin in preventing strokes, with a relative risk reduction of 36%, but the risk of major hemorrhage with warfarin was twice that with aspirin. Anticoagulation treatment needs to be tailored individually for patients on the basis of age, comorbidities, and contraindications. However, warfarin remains under-prescribed in clinical practice, for reasons related to patients (comorbidities) and physicians. The limitations of warfarin treatment have prompted the development of new anticoagulants with predictable pharmacokinetics that do not require as frequent monitoring. Ximelagatran, an oral direct thrombin inhibitor, was compared with warfarin in the SPORTIF program, which found both agents to be broadly effective in the prevention of embolic events, but observed abnormal liver function tests in 6% of patients on ximelagatran. Liver function monitoring during treatment is thus needed. Idraparinux, a factor Xa inhibitor administered by once weekly subcutaneous injections, is being evaluated in patients with atrial fibrillation. The ACTIVE trial is currently assessing the role of aspirin plus clopidogrel, compared with adjusted dose warfarin, in the prevention of vascular events in high-risk patients with atrial fibrillation. Angiotensin-converting enzyme inhibitors and angiotensin II receptor-blocking drugs interfere with atrial remodeling and show promise in atrial fibrillation, as suggested in the LIFE trial. Preliminary studies suggest that statins may reduce the risk of recurrence after electrical cardioversion. Finally, percutaneous methods for occlusion of the left atrial appendage are currently under investigation in patients at high risk of thromboembolism but with contraindications for chronic warfarin. PMID- 16269997 TI - New anticoagulants in ischemic heart disease. AB - Fibrinolysis is the reference treatment for most myocardial infarctions with ST segment elevation; alternatives are angioplasty, with or without stent. The earlier fibrinolysis is performed (preferably before hospitalization), the more effective it is. It can be optimized by adjuvant antiplatelet therapy, such as aspirin, and probably by anticoagulant treatment as well. Because fibrinolytic therapy is accompanied by intensive thrombin generation and activation, immediate and continuous adjunctive simultaneous heparin therapy is recommended. The efficacy of subcutaneous low-molecular-weight heparin (LMWH) HBPM) is at least equivalent to that of intravenous unfractionated heparin (UFH), but its risk of severe (but not cerebral) hemorrhage is greater. Bolus LMWH on the other hand is associated with an increased risk of cerebral hemorrhage. Antithrombotic treatment thus appears optimal with bolus UFH at fibrinolysis and for at least 48 hours afterwards. An alternative after this bolus might be subcutaneous enoxaparin until discharge. Because the major drawback of both types of heparin is their rebound activation of thrombosis, oral anticoagulants are recommended thereafter. The combination of anticoagulant treatment + (low-dose) aspirin is not superior to aspirin alone when the target INR is below 2. Adequate anticoagulation with INRs greater than 2.0 consistently improves angiographic and clinical outcome. Bleeding (except intracerebral) is significantly increased whether the INR is greater than or less than 2.0. Other treatments are being investigated. Pentasaccharide (anti-Xa) combined with fibrinolysis seems to reduce reocclusion more effectively than UFH. Oral postinfarction treatment with ximelagatran (a thrombin inhibitor), combined with aspirin, is associated with fewer cardiovascular events than aspirin alone. More studies are needed. PMID- 16269999 TI - [Recurrent serpentine thrombus of the right cardiac chambers]. PMID- 16269998 TI - [Medications in smoking cessation]. AB - Physicians can aid their patients' smoking cessation by providing psychological support, advice, behavioral strategies, and drugs. Success depends on appropriate management, including selection of the right moment to begin treatment and an understanding of the development of the withdrawal syndrome, smoking urges, and the possibility of failure. The standard pharmacological treatment for nicotine dependence uses different forms of nicotine substitutes and bupropion, while we await data about other drugs currently under study. The score on the "simplified" Fagerstrom questionnaire usually determines the initial nicotine dose. Six forms of nicotine substitutes are available. They provide either prolonged nicotine release (transcutaneous patches) that prevents withdrawal symptoms, or rapid release through the buccal and nasal mucosa (chewing gum, suckers, inhalers and nasal sprays) to anticipate the positive effects represented by cigarettes and the urges occurring during withdrawal. The efficacy of these substitutes, widely studied, is approximately twice that of placebo. Their use is no longer contraindicated in patients with heart disease, when necessary. Bupropion should be used in treating nicotine dependence either as a first-line treatment, or if nicotine substitutes (150 mg/d the first week, 300 mg/d thereafter) fail. The combination of bupropion and nicotine substitutes can be considered, either from the outset for heavy or very heavy smokers, or afterwards, if withdrawal symptoms or urges to smoke persist in subjects treated by only one of these two drug classes. One of the new drugs under evaluation is rimonabant, the first representatives of a new class of drugs, selective CB1 endocannabinoid receptor antagonists. Promising results about its use in smoking cessation were released in 2004. PMID- 16270000 TI - [The gynecology-obstetrics pole option: a major risk or a plus for our specialty?]. PMID- 16270001 TI - [Adenomyosis: update on a frequent but difficult diagnosis]. AB - Adenomyosis is a frequent entity, with difficult diagnosis, often obtained by pathological analysis performed after hysterectomy. This condition can cause abnormal uterine bleeding and dysmenorrhea, frequent reasons for consultation and hysterectomy. The development of ultrasonographic and magnetic resonance imaging techniques allow preoperative diagnosis. They also permit the use of hysteroscopic techniques for conservative uterine surgery, and have brought diagnosis and management of this disease to the front of the scene. This article reviews the pathological description of the disease, its epidemiology, clinical presentations, useful and necessary explorations, etiopathogeny and available therapies. PMID- 16270002 TI - [Ovarian autoimmunity and ovarian pathologies: antigenic targets and diagnostic significance]. AB - The involvement of serum anti-ovarian autoantibodies (AOA) in ovarian pathology still remains controversial. In some cases of clinically patent ovarian failure, there seems to be a causal relationship between AOA and the ovarian disease. In patients with various organ-specific or systemic autoimmune diseases, or with unexplained, repeated reproductive failure, but otherwise normal ovarian function, it is even more difficult to determine the significance of AOA for several reasons: i) AOA recognize many different antigenic targets in the ovary ii) the antiovarian response may be transient or variable with time iii) the presence of AOA does not imply their aetiopathogenic role in the disease. The present paper reviews the clinical significance of AOA based on their ovarian targets as far as they have been identified until now. PMID- 16270003 TI - [Methodology of the inquiry MISSION (menopause, breast cancer risk, morbidity and prevalence)]. AB - OBJECTIVE: The ambition of this article is to detail and to explain the methodology of the study named MISSION (Menopause: breast cancer risk, morbidity and prevalence). The aims of MISSION are to determine the prevalence of breast cancer and global morbidity in France among menopausal women treated or not with hormone replacement therapy (HRT) and followed by a gynecologist. MATERIAL AND METHOD: 6600 menopausal women [3300 with HRT -- ie for estrogen: only estradiol via oral or transdermal administration; for progestogen: natural progesterone or assimiled or one pregnane derived (excluding medroxyprogesterone acetate) or non pregnane derived -- and 3300 without HRT] will be enrolled in France between January 5 2004 and February 28 2005 by 825 gynecologists, members or not of the National Federation of Medical Gynecologists (FNCGM). This study design is a historico-prospective with case randomization. MISSION is conducted by a Theramex Merck Laboratories initiative in collaboration with a WHO (World Health Organization) Collaborating Center for Public Health Aspects of Rheumatic Diseases and a multidisciplinary expert group. CONCLUSION: First results of this study will contribute to better knowledge of women health. PMID- 16270004 TI - [Placental functional assessment using MRI: mice today, humans tomorrow?]. AB - Placental insufficiency, a process due to either poor placental perfusion or permeability, may lead to progressive deterioration in placental function and materno-fetal morbidity. Advances in MR contrast media pharmacokinetic studies of transit through tissues and dynamic MRI allow to characterize organs microcirculation in vivo. Placental function assessment might be achieved using analysis of dynamic contrast enhanced MRI of tracers. A murine model of placental assessment has been constructed. Herein, principles, results and limitations of such techniques are discussed as well as their potential interest and weaknesses in humans. PMID- 16270005 TI - [Obstetrical ultrasound screening and motivation to stop smoking]. AB - OBJECTIVES: To evaluate whether ultrasound screening during pregnancy increases the ability to stop or reduce smoking. MATERIALS AND METHODS: This multicenter questionnaire study was performed to describe smoking characteristics among pregnant women and evaluate the factors that influence the ability to reduce smoking during pregnancy. Questionnaire-based, descriptive study of 979 pregnant women in four regions of France. The variables analyzed included the characteristics of the mother and neonate at delivery, the smoking habits of the mother before and during pregnancy, and the reasons for giving up smoking. RESULTS: We report only results about ultrasound scan. One third of women, who smoked at the start of pregnancy, reported that ultrasound was a positive tool to enhance motivation to reduce smoking. CONCLUSION: Ultrasound screening may be a positive factor to reduce smoking. Our study focused on motivation could give a tool in this way. However most women who stop smoking during pregnancy do so in the first trimester (84%) mainly before the first ultrasound exam. Thus, the routine ultrasound scan itself does not seem to influence attitudes to stop smoking any further. But it may be included in comprehensive and individualised anti-smoking support. PMID- 16270006 TI - [Nicotine replacement therapies during pregnancy: what do we know about the balance between benefits and risks?]. AB - Maternal smoking during pregnancy induces obstetrical and fetal complications but also has an impact on newborns, infants, children and adults. Nicotine replacement therapies are authorized during pregnancy in France, the purpose being to reduce fetal exposure to toxic compounds in tobacco smoke. However, it is not proven that nicotine replacement therapy is harmless to the fetus and to date, no study has demonstrated any beneficial effect in terms of abstinence. It is suggested that benefit and risks of nicotine replacement therapies during pregnancy should be evaluated. PMID- 16270007 TI - [Comparison of women's alcohol consumption before and during pregnancy. A prospective series of 150 women]. AB - OBJECTIVES: Evaluation of the prevalence of prenatal alcohol exposure in relation with declared alcohol consumption (DAC). MATERIALS AND METHODS: Observational prospective study based on the DAC of 150 women in post-partum or hospitalized for abnormal pregnancies. The main selection criterion was change in DAC before and during pregnancy, classified according to alcohol use and misuse limits. Excessive alcohol intake was defined as more than 4 glasses on occasion and/or more than 14 glasses a week. Age and professional position were also taken into account. Data were obtained using semi-directive interviews. RESULTS: 79% of these women drank before pregnancy. 33.2% of them had excessive intake. Consumption of more than 14 glasses a week concerned 10% of the population. Excessive consumptions on occasion significantly affected 33% of the population. During pregnancy, 43% of these women continued drinking. Overall excessive intake, which is noxious for children, concerned 9.9% of the women. 5% declared a regular misuse above 14 glasses a week and 7% of them consumed more than 4 glasses on occasion. The average age was 29.8 years. Most of the women (61%) had a stable job. CONCLUSION: Our study confirms alcohol consumption by women, even during pregnancy. Consumptions of pregnant women is decreasing progressively. DAC is very useful to evaluate alcohol intake. PMID- 16270008 TI - [Obstetrical risk factors of 365 primiparous adolescent pregnancies in Reunion Island]. AB - OBJECTIVES: Analysis of obstetrical risk factors in teenage primiparous pregnancies in Reunion Island (4% of total births). MATERIALS AND METHODS: Retrospective study, between 2001 and 2002, comparing primiparous adolescents (13 17 years, n = 365), with primiparous controls (18-29 years, n = 2050). The analysis included demographical factors, maternal medical histories, prenatal follow-up, obstetrical risk factors, delivery modes and neonatal characteristics. RESULTS: Adolescents attended on average 8 prenatal consultations, however 4% had poor prenatal care (less than 3 visits, OR 4.2, P < 0.001 vs controls). They presented less gestational diabetes, but there were no differences concerning pre existing hypertension, hypertensive disorders of pregnancy, medical reasons of hospitalisation between the two groups. Mode of delivery was more favorable in adolescents (half rate of caesarean sections, shorter duration of membrane ruptures). Nevertheless, adolescents presented a higher risk of severe prematurity (<32 Weeks gestation, incidence 3.6% vs 1.6%, OR 2.3, p = 0.008). CONCLUSION: With optimal prenatal care (more than 90% of our cohort), primiparous adolescents present globally a favorable course of their pregnancies and have better deliveries than their young (18-29 years) counterparts. However, there is a significant risk of severe prematurity requiring special care for these pregnancies. PMID- 16270010 TI - [Maternal diaphragmatic hernia during pregnancy: report of two cases]. AB - We report two cases of maternal diaphragmatic hernia during pregnancy. Diaphragmatic hernia is an unusual and severe disease. Maternal and fetal prognosis are threatened. Diagnosis is uncertain when confronted to respiratory and digestive symptoms without any specificity. The chest X ray is the first exam to perform. The objective of this work is to discuss the management of such a pathology in terms of ways of delivery and surgical cure of hernia. PMID- 16270009 TI - [Transurethral injections under local anaesthesia for ambulatory treatment of stress urinary incontinence in women: indications, feasibility and results]. AB - OBJECTIVE: The aim of the study was to evaluate feasibility and results of ambulatory transurethral injections of Macroplastique using the system MIS under local anaesthesia, for treatment of female stress urinary incontinence. MATERIAL AND METHODS: We have performed a prospective multicentre study which has enrolled 20 patients between January 2003 and May 2004. Mean age was 72.8 ans (range 40 to 91). Preoperative inclusion criterias were positive stress test, 24 hours PAD test more than 10 g and post-void residual less than 100 ml. Urethral hypermobility was present in 8 patients (42.1%), and 3 patients was diagnosed as intrinsic sphincter deficiency (16.7%) on urodynamics examination. Pre and post operative evaluation of urinary symptoms and quality of life were performed with the MHU score (Mesure du Handicap Urinaire), the Ditrovie score and the Contilife scale. The Macroplastique implant (Uroplasty) contains silicone and a bioabsorbable gel. Injections were perfomed under the mid-urethral mucosa (2.5 ml at 6 hours and 1.25 ml at 2 and 10 hours) using the system MIS (Uroplasty). No cystoscopy was required and the mean operative time was 15 minutes. RESULTS: The procedure was feasible under pure local anaesthesia in all cases. No intra operative complications occurred. Postoperative complications had included 2/19 local pain (10.5%), 3/19 minimal urethrorrhagia (15.8%) and 6/19 urinary retentions (31.6%), which were treated by heterologous intermittent catheterization during 3 to 20 days. Eight patients returned home the same day (42.1%). The mean hospital stay was 2.3 days. Mean postoperative follow-up was 8.3 months (range 2.7 to 19.1). No patient required a second injection. Results had shown a 36.9% success rate (7/19), 52.6% improvement (10/19) and 10.5% failure (2/19). At follow-up, stress test was negative in 66.7% of patients (12/18) and PAD test was<10 g in 66.7% (10/15). Modifications on maximal flow rate were non significant after injections. Postoperatively, all urinary symptoms were improved except nocturia and voiding difficulties. Quality of life was improved on all parameters. CONCLUSION: The surgical treatment of female stress urinary incontinence by transurethral injections of Macroplastique using the system MIS under local anaesthesia was feasible in all cases with a success or improvement rate of 89.5% at a mean follow-up of 8.3 months. Ambulatory treatment was compromised by the high rate of postoperative urinary retention (31.6%). PMID- 16270012 TI - [Primary non-Hodgkin's lymphomas of the breast. Report of two cases]. AB - Non-hodgkin's lymphoma (NHL) represents 0.04 to 0.53% of all breast cancers. The clinical aspects and therapeutic models of the disease are a subject of debate. We report two cases of primary non-hodgkin's lymphoma of the breast in two patients aged respectively 37 and 57 years. The disease was revealed by breast nodules. Mammography demonstrated a benign cyst. The diagnosis of non-hodgkin's lymphoma was confirmed on histological examination of tumor biopsies. With chemotherapy, the course was favorable with 29 and 52 months follow-up respectively. This is rare tumor. The diagnosis is mainly histological. Chemotherapy is the principal therapeutic method. PMID- 16270011 TI - [Cystic fibrosis and pregnancy: report of a twin pregnancy and review of the literature]. AB - We report here the first published case of twin pregnancy in a woman with cystic fibrosis. This situation will become more and more common because the increased life expectancy of patients with cystic fibrosis and the development of medically assisted procreation. Conception of this twin pregnancy was medically assisted in a cystic fibrosis woman with moderate pulmonary disease. This observation is in accordance with several recent reports dealing with single pregnancies: morbidity, mortality and degradation of lung function have not been found to increase. The only effect of cystic fibrosis on pregnancy is an increased risk of preterm delivery. There is thus no systematic contraindication to pregnancy among women with cystic fibrosis provided that they are given attentive care by a multidisciplinary team. Genetic counselling and prenatal diagnosis also have an important role to play. PMID- 16270013 TI - [Evaluation of an absorbed dose to the human body by measuring ionizations in a cavity chamber]. PMID- 16270014 TI - [How to apply X-ray CT reasonably: obligation of the radiologists]. PMID- 16270015 TI - [The point of mammography]. PMID- 16270016 TI - [Strategy of evidence based medicine for radiotherapy]. PMID- 16270017 TI - [This and that in construction of systems]. PMID- 16270018 TI - [The consideration about the storage selection at the time of system introduction, and a user to be concerned]. PMID- 16270020 TI - [Hokkaido Helical CT Research Group]. PMID- 16270019 TI - [The characteristic of a photo luminescence dosimeter: the scope of radiotherapy]. PMID- 16270021 TI - [Economical benefit of diagnostic imaging: part 1]. PMID- 16270022 TI - [Development of a breath pacemaker for 2D-PACE respiratory gated MRCP: efficacy of image quality improvement]. AB - Two-dimensional prospective acquisition correction (2D-PACE) is a human body motion-correction technique that uses the navigator echo sequence. Clinically, this respiratory-gated technique is used to improve the image quality of magnetic resonance cholangiopancreatography (MRCP). However respiratory motion artifact caused by a mismatch in respiratory rhythm has been experienced. The newly developed breath pacemaker was developed with the aim of providing the best match between the breath cycle and 2D-PACE trigger. The breath pacemaker is composed of two stimulators, an auto voice (AV) system and an auto light (AL) system. The two systems can be controlled separately or concurrently. Using the new breath pacemaker, improvement in MRCP image quality was evaluated in normal volunteers, and definite improvement was noted. Particularly good efficacy was obtained with concurrent control of the AV system and AL system. The results clearly demonstrated that the breath pacemaker can provide good synchronization of 2D PACE and can lead to enhanced quality of MRCP images. PMID- 16270023 TI - [Optimization of flip angle for 3D-VIBE technique in liver dynamic MRI]. AB - The aim of this study was to determine a moderate flip angle (FA) for dynamic liver MR imaging (MRI) with the three-dimensional volumetric interpolated breath hold examination (3D-VIBE) technique. Images of phantoms with various T(1) values (44-560 msec) were acquired with the 3D-VIBE sequence (TR/TE=5.2/2.6 msec) using different FA (5-50 degree). We estimated signal-to-noise ratio (SNR) and contrast to-noise ratio (CNR), considered to indicate tumor-to-liver contrast, as a function of FA. In phantoms, in which T(1) values (44-191 msec) were assumed to be shortened by the effect of Gd-DTPA in hepatocellular carcinoma (HCC), the highest SNR in each phantom was observed at FA ranging from 15 to 30 degrees. SNRs in other phantoms, in which T(1) values (298-560 msec) were assumed to be normal liver-tissue pre- or post-enhancement, were high with FA of 10-12 degrees, and were remarkably decreased with FA of more than 30 degrees. CNR increased as FA increased in every phantom, especially in the phantom with shortened T(1) values (44-191 msec), suggesting that enlarging FA improved the tumor-to-liver contrast. Taking both results of SNR and CNR into account, we concluded that a moderate FA was approximately 25 degrees. The advantage with an FA of 25 degrees was confirmed in a clinical study of a patient with hypervascular HCC, in which we could observe coronal enhancement surrounding the lesion in the late phase of the double arterial phase by dynamic MRI using the 3D-VIBE technique. PMID- 16270024 TI - [Examination of retention index in brain tumor 201Tl scintigraphy using iNRT]. AB - The retention index (R.I.) in Thalliun-201-chloride (201Tl-Cl) brain tumor single photon emission computed tomography (SPECT) imaging analysis is useful in assessing grades of malignancy. When the T/N ratio that influences the R.I. is calculated, it is ideal to draw an ROI on the same slices and coordinates of both early and delayed transaxial images. However, in many institutions, the image slice and ROI are determined manually by operators (i.e., by the conventional method). The Co-registration function within iNRT software enables an ROI to be automatically drawn. In this study, we obtained the R.I. from six brain tumor patients employing two different methods (i.e., Co-registration and the conventional method). Each R.I. was obtained from the same personal computer that was used to transfer the transaxial images. We then compared and evaluated the six R.I. values of seven operators. (2 radiologists and 5 radiological technologists). The results showed that the R.I. values did not differ significantly between the two methods, except in two cases. Furthermore, the coefficient of variation (CV) of the R.I. obtained from the Co-registration method was lower than the conventional method in all cases. The variations among operators were also smaller in the Co-registration method. These outcomes may have been due to the fact that in the conventional method it is difficult to identify the same slices of early and delayed images. Therefore, there were differences among operators in their ways of specifying positions of the ROI. This may have resulted in differences in the number of counts/pixel of the ROI. In conclusion, the R.I. obtained by using the Co-registration method would be more useful for improving diagnostic accuracy in (201)Tl-Cl brain tumor SPECT imaging than the conventional method. PMID- 16270025 TI - Differential conversion of plasminogen to angiostatin by human corneal cell populations. AB - PURPOSE: Maintenance of avascularity of the normal cornea and control of neovascularization during wound healing depend on a balance of angiogenic and antiangiogenic factors. The purpose of this paper is to determine the ability of corneal cells to convert plasminogen to angiostatins and to compare these products with those made by intact corneas. METHODS: RT-PCR was performed using plasminogen specific primers and the generated cDNA was sequenced. The proteins in corneal extracts, cornea conditioned medium, and medium from corneal epithelial cells, stromal fibroblasts, and myofibroblasts incubated with plasminogen were separated by SDS-PAGE and electroblotted. Western blots used monoclonal antibodies to kringles 1-3 to detect plasminogen and angiostatins. Angiostatins were isolated and tested for activity in a vascular endothelial cell proliferation inhibition assay. RESULTS: Plasminogen, its mRNA and angiostatins were found in human corneal tissue extracts from the epithelial, stromal, and endothelial layers and from cornea conditioned medium, but not in medium from cultured epithelial cells, stromal fibroblasts, or myofibroblasts. However, cultures of corneal epithelial cells and stromal fibroblasts were able to convert exogenously added plasminogen to angiostatins, whereas cultured myofibroblasts did not. Angiostatins of 38 and 34 kDa were found under all angiostatin generating conditions; however other angiostatins differed in size. Further, the angiostatins isolated from fibroblast culture supernatants inhibited vascular endothelial cell proliferation. CONCLUSIONS: Conversion of plasminogen to angiostatin is cell-type dependent. Because corneal cells generate angiostatins, use of human angiostatins may be a means of treating abnormal corneal neovascularization without the risk of side effects. PMID- 16270026 TI - Barriers to productive transfection of trabecular meshwork cells. AB - PURPOSE: A critical function of trabecular meshwork cells is to degrade cellular debris, including DNA. We hypothesize that low transfection efficiencies of primary human trabecular meshwork (HTM) cell cultures with plasmid DNA are a function of retained capacity to efficiently degrade exogenous DNA in vitro. METHODS: To determine mechanisms responsible for low transfection efficiencies of cultured HTM cells, steps of DNA entry into cytoplasm and nucleus were characterized. Following synchronization with sequential serum starvation and serum reintroduction, the HTM cell cycle was characterized using 5-bromo-2 deoxyuridine incorporation into replicating DNA. HTM cells were transfected during S-phase with plasmid DNA encoding green fluorescence protein (GFP) or plasmid DNA conjugated with Cy3. In some experiments, cells were treated with a DNase I inhibitor, 100 nM aurintricarboxylic acid. Uptake of plasmid DNA was measured by intracellular fluorescence of Cy3 and productive transfection efficiency was measured by intracellular fluorescence of GFP. RESULTS: HTM cells enter S-phase between 18 and 20 h after synchronization. Plasmid DNA reached the cytosolic compartment in 95% of transfected cells, regardless of synchronization. Synchronization dramatically increased productive transfection efficiency in HTM cells, from 3.0 to 9.0%. DNase I inhibition increased productive transfection efficiency of HTM cells two fold. CONCLUSIONS: Cultured HTM cells have a lower transfection efficiency than other primary ocular cell cultures, likely due partially to cytoplasmic digestion of DNA. We suggest that the difficulties in transfecting cultured HTM cells may be related to the filter function of the cells in vivo where the cells must degrade exogenous DNA. PMID- 16270027 TI - Intracorneal positioning of the lens in Pax6-GAL4/VP16 transgenic mice. AB - PURPOSE: The purpose of this study was to establish a GAL4/VP16-based binary transactivation system that was active in the lens and corneal epithelium of transgenic mice. METHODS: We generated transgenic mice with the transcriptional transactivator GAL4/VP16 driven by a modified Pax6 promoter that is active in lens and corneal epithelial cells. We also generated and tested UAS-lacZ reporter mice. Wild type and transgenic mice were analyzed by histological, in situ, and Southern hybridization techniques. RESULTS: Five families (OVE1931, OVE1934, OVE1935, OVE1936, and OVE1937) that carry the Pax6-GAL4/VP16 transgene were generated. Unexpectedly, mice from three of the transgenic lines showed ocular abnormalities. In the family OVE1936, cataracts were seen in the heterozygous mice at the time of eyelid opening and homozygotes showed microphthalmia. Transgenic mice in families OVE1931 and OVE1937 appeared normal. Histological analysis of ocular sections of OVE1934, OVE1935, and OVE1936 homozygous transgenic mice showed intracorneal positioning of the lens. The corneal stromal cells were disorganized and there was no distinctive corneal endothelial layer. In situ hybridizations showed robust expression of the GALVP16 transgene in the lens and corneal epithelial cells of the OVE1934, OVE1935, and OVE1936, but not in OVE1931 or OVE1937 families. Bigenic embryos generated by mating the Pax6 GAL4/VP16 mice to the UAS-lacZ mice showed that the GAL4/VP16 transgenic protein is functional and can induce eye-specific expression of a UAS-lacZ reporter gene. CONCLUSIONS: Our data suggest that (1) expression the GAL4/VP16 transgene induces changes in gene expression in lens cells, (2) that developmentally important genes are affected, and (3) that bigenic phenotypes will need to be interpreted with caution. PMID- 16270028 TI - Activation of microglia and chemokines in light-induced retinal degeneration. AB - PURPOSE: Microglial cells, which are activated and recruited by chemokines, have been shown to play crucial roles in neuronal degenerations of the central nervous system (CNS). This study investigated the activation and migration of retinal microglial cells and expression of chemokines in retinas in light-induced photoreceptor degeneration in mice. METHODS: Ninety-five Balb/cJ mice were kept in cyclic light for 1 week followed by dark adaptation for 48 h prior to light exposure of 3 h at 3.5 Klux. Animals were enthuanized at various times after light exposure. Terminal deoxynucleotidyl transferase-mediated dUTP nick end label (TUNEL) assay, rat-anti-mouse CD11b and 5D4 antibodies, isolectin-B4, and a chemokine-specific gene array were used to detect DNA fragmentation during retinal degeneration, to label retinal microglial cells, and to determine the expression of retinal chemokines and chemokine receptors, respectively. Reverse transcriptase coupled polymerase chain reactions (RT-PCRs) were conducted on selected chemokine mRNAs to confirm the gene array findings. RESULTS: After intense light exposure, TUNEL-positive cells were noted in the outer nuclear layer (ONL) of the retina at 3 h, and their presence were noticeably increased at 1 day but declined at 3 days and 7 days after light exposure. In contrast, CD11b- or isolectin-B4-positive cells were seen in the ONL as early as 6 h and their presence increased significantly at 1 day and 3 days after light exposure. These cells displayed a round or ovoid morphology at 6 h and 1 day but assumed a more ameboid configuration at 3 days. By 7 day, the number of the microglial cells declined in the ONL and they became ramified, and were present mostly in the subretinal space. 5D4-positive cells with large cell bodies were only noted at 3 day and 7 day but not earlier. With chemokine-specific gene array analysis, we identified four chemokines and two chemokine receptors showing significant increases in their gene expressions. Among them, monocyte chemoattractant protein 3 (MCP-3), showed a remarkable 4.4 fold increase in its gene expression. RT-PCR confirmed a marked increase of MCP-3 expression in retinas at 3 h to 1 day, and a return to normal at 3 days following light injury. CONCLUSIONS: Retinal chemokines such as MCP-3 and their receptors are involved in the activation and migration of retinal microglia in light-induced retinal degeneration, which in turn modulate the apoptotic loss of photoreceptor cells in the outer retina. PMID- 16270029 TI - Mobilization of vacuolar iron by AtNRAMP3 and AtNRAMP4 is essential for seed germination on low iron. AB - Iron (Fe) is necessary for all living cells, but its bioavailability is often limited. Fe deficiency limits agriculture in many areas and affects over a billion human beings worldwide. In mammals, NRAMP2/DMT1/DCT1 was identified as a major pathway for Fe acquisition and recycling. In plants, AtNRAMP3 and AtNRAMP4 are induced under Fe deficiency. The similitude of AtNRAMP3 and AtNRAMP4 expression patterns and their common targeting to the vacuole, together with the lack of obvious phenotype in nramp3-1 and nramp4-1 single knockout mutants, suggested a functional redundancy. Indeed, the germination of nramp3 nramp4 double mutants is arrested under low Fe nutrition and fully rescued by high Fe supply. Mutant seeds have wild type Fe content, but fail to retrieve Fe from the vacuolar globoids. Our work thus identifies for the first time the vacuole as an essential compartment for Fe storage in seeds. Our data indicate that mobilization of vacuolar Fe stores by AtNRAMP3 and AtNRAMP4 is crucial to support Arabidopsis early development until efficient systems for Fe acquisition from the soil take over. PMID- 16270030 TI - The F1-ATP synthase complex in bloodstream stage trypanosomes has an unusual and essential function. AB - Survival of bloodstream form Trypanosoma brucei, the agent of African sleeping sickness, normally requires mitochondrial gene expression, despite the absence of oxidative phosphorylation in this stage of the parasite's life cycle. Here we report that silencing expression of the alpha subunit of the mitochondrial F(1) ATP synthase complex is lethal for bloodstream stage T. brucei as well as for T. evansi, a closely related species that lacks mitochondrial protein coding genes (i.e. is dyskinetoplastic). Our results suggest that the lethal effect is due to collapse of the mitochondrial membrane potential, which is required for mitochondrial function and biogenesis. We also identified a mutation in the gamma subunit of F(1) that is likely to be involved in circumventing the requirement for mitochondrial gene expression in another dyskinetoplastic form. Our data reveal that the mitochondrial ATP synthase complex functions in the bloodstream stage opposite to that in the insect stage and in most other eukaryotes, namely using ATP hydrolysis to generate the mitochondrial membrane potential. PMID- 16270034 TI - PKCepsilon-mediated phosphorylation of vimentin controls integrin recycling and motility. AB - PKCepsilon controls the transport of endocytosed beta1-integrins to the plasma membrane regulating directional cell motility. Vimentin, an intermediate filament protein upregulated upon epithelial cell transformation, is shown here to be a proximal PKCepsilon target within the recycling integrin compartment. On inhibition of PKC and vimentin phosphorylation, integrins become trapped in vesicles and directional cell motility towards matrix is severely attenuated. In vitro reconstitution assays showed that PKCepsilon dissociates from integrin containing endocytic vesicles in a selectively phosphorylated vimentin containing complex. Mutagenesis of PKC (controlled) sites on vimentin and ectopic expression of the variant leads to the accumulation of intracellular PKCepsilon/integrin positive vesicles. Finally, introduction of ectopic wild-type vimentin is shown to promote cell motility in a PKCepsilon-dependent manner; alanine substitutions in PKC (controlled) sites on vimentin abolishes the ability of vimentin to induce cell migration, whereas the substitution of these sites with acidic residues enables vimentin to rescue motility of PKCepsilon null cells. Our results indicate that PKC-mediated phosphorylation of vimentin is a key process in integrin traffic through the cell. PMID- 16270031 TI - Concomitant loss of proapoptotic BH3-only Bcl-2 antagonists Bik and Bim arrests spermatogenesis. AB - The BH3-only proteins of the Bcl-2 family initiate apoptosis through the activation of Bax-like relatives. Loss of individual BH3-only proteins can lead either to no phenotype, as in mice lacking Bik, or to marked cell excess, as in the hematopoietic compartment of animals lacking Bim. To investigate whether functional redundancy with Bim might obscure a significant role for Bik, we generated mice lacking both genes. The hematopoietic compartments of bik-/-bim-/- and bim-/- mice were indistinguishable. However, although testes develop normally in mice lacking either Bik or Bim, adult bik-/-bim-/- males were infertile, with reduced testicular cellularity and no spermatozoa. The testis of young bik-/-bim /- males, like those lacking Bax, exhibited increased numbers of spermatogonia and spermatocytes, although loss of Bik plus Bim blocked spermatogenesis somewhat later than Bax deficiency. The initial excess of early germ cells suggests that spermatogenesis fails because supporting Sertoli cells are overwhelmed. Thus, Bik and Bim share, upstream of Bax, the role of eliminating supernumerary germ cells during the first wave of spermatogenesis, a process vital for normal testicular development. PMID- 16270035 TI - Major molecular response to imatinib in a patient with chronic myeloid leukemia expressing a novel form of e8a2 BCR-ABL transcript. PMID- 16270032 TI - INSIG: a broadly conserved transmembrane chaperone for sterol-sensing domain proteins. AB - INSIGs are proteins that underlie sterol regulation of the mammalian proteins SCAP (SREBP cleavage activating protein) and HMG-CoA reductase (HMGR). The INSIGs perform distinct tasks in the regulation of these effectors: they promote ER retention of SCAP, but ubiquitin-mediated degradation of HMGR. Two questions that arise from the discovery and study of INSIGs are: how do they perform these distinct tasks, and how general are the actions of INSIGs in biology? We now show that the yeast INSIG homologs NSG1 and NSG2 function to control the stability of yeast Hmg2p, the HMGR isozyme that undergoes regulated ubiquitination. Yeast Nsgs inhibit degradation of Hmg2p in a highly specific manner, by directly interacting with the sterol-sensing domain (SSD)-containing transmembrane region. Nsg1p functions naturally to limit degradation of Hmg2p when both proteins are at native levels, indicating a long-standing functional interplay between these two classes of proteins. One way to unify the known, disparate actions of INSIGs is to view them as known adaptations of a chaperone dedicated to SSD-containing client proteins. PMID- 16270036 TI - Cells enriched in markers of neural tissue-committed stem cells reside in the bone marrow and are mobilized into the peripheral blood following stroke. AB - The concept that bone marrow (BM)-derived cells participate in neural regeneration remains highly controversial and the identity of the specific cell type(s) involved remains unknown. We recently reported that the BM contains a highly mobile population of CXCR4+ cells that express mRNA for various markers of early tissue-committed stem cells (TCSCs), including neural TCSCs. Here, we report that these cells not only express neural lineage markers (beta-III tubulin, Nestin, NeuN, and GFAP), but more importantly form neurospheres in vitro. These neural TCSCs are present in significant amounts in BM harvested from young mice but their abundance and responsiveness to gradients of motomorphogens, such as SDF-1, HGF, and LIF, decreases with age. FACS analysis, combined with analysis of neural markers at the mRNA and protein levels, revealed that these cells reside in the nonhematopoietic CXCR4+/Sca-1+/lin-/CD45 BM mononuclear cell fraction. Neural TCSCs are mobilized into the peripheral-blood following stroke and chemoattracted to the damaged neural tissue in an SDF-1-CXCR4-, HGF-c-Met-, and LIF-LIF-R-dependent manner. Based on these data, we hypothesize that the postnatal BM harbors a nonhematopoietic population of cells that express markers of neural TCSCs that may account for the beneficial effects of BM-derived cells in neural regeneration. PMID- 16270037 TI - Myeloablative vs nonmyeloablative allogeneic transplantation for patients with myelodysplastic syndrome or acute myelogenous leukemia with multilineage dysplasia: a retrospective analysis. AB - Transplant outcome was analyzed in 150 patients with myelodysplastic syndrome (MDS) or acute myelogenous leukemia transformed from MDS (tAML) conditioned with nonmyeloablative or myeloablative regimens. A total of 38 patients received nonmyeloablative regimens of 2 Gy total body irradiation alone (n=2) or with fludarabine (n=36), 90mg/m2. A total of 112 patients received a myeloablative regimen of busulfan, 16mg/ kg (targeted to 800-900 ng/ml), and cyclophosphamide 120 mg/ kg. Nonmyeloablative patients were older (median age 62 vs 52 years, P<0.001), more frequently had progressed to tAML (53 vs 31%, P=0.06), had higher risk disease by the International Prognostic Scoring System (53 vs 30%, P=0.004), had higher transplant specific comorbidity indices (68 vs 42%, P=0.01) and more frequently had durable complete responses to induction chemotherapy (58 vs 14%). Three-year overall survival (27%/48% (P=0.56)), progression-free survival (28%/4 44%, (P=0.60)), and nonrelapse mortality (41%/34%, (P=0.94)) did not differ significantly between nonmyeloblative/myeloablative conditioning. Overall (HR=0.9, P=0.84) and progression-free survivals (HR=1, P=0.93) were similar for patients with chemotherapy-induced remissions irrespective of conditioning intensity. Graft vs leukemia effects may be more important than conditioning intensity in preventing progression in patients in chemotherapy-induced remissions at the time of transplantation. Randomized prospective studies are needed to further address the optimal choice of transplant conditioning intensity in myeloid neoplasms. PMID- 16270038 TI - Cyclin D2 dysregulation by chromosomal translocations to TCR loci in T-cell acute lymphoblastic leukemias. AB - Strong expression of at least one of the three D-type cyclins is common in human cancers. While the cyclin D1 and D3 genes (CCND1 and CCND3) are recurrently involved in genomic rearrangements, especially in B-cell lymphoid neoplasias, no clear involvement of the cyclin D2 gene (CCND2) has been reported to date. Here, we identified chromosomal translocations targeting the CCND2 locus at 12p13, and the T-cell receptor beta (TCRB) or the TCRA/D loci in T-cell acute lymphoblastic leukemias (T-ALLs). Expression analysis demonstrated dramatic cyclin D2 overexpression in the translocated cases (n=3) compared to other T-ALLs (total, n=89). In order to evaluate dysregulation in T-ALL with respect to normal T-cell differentiation, we analyzed CCND2 expression in normal purified human thymic subpopulations. CCND2 levels were downregulated through progression from the early stages of human T-cell differentiation, further suggesting that the massive and sustained expression in the CCND2-rearranged T-ALL cases was oncogenic. Association with other oncogene expression (TAL1, HOXAs, or TLX3/HOX11L2), NOTCH1 activating mutations, and/or CDKN2A/p16/ARF deletion, showed that cyclin D2 dysregulation could contribute to multi-event oncogenesis in various T-ALL groups. This report is the first clear evidence of a direct involvement of cyclin D2 in human cancer due to recurrent somatic genetic alterations. PMID- 16270040 TI - Acute leukaemia in adults: researching the patient's perspective. PMID- 16270039 TI - Validation of two clinically useful assays for evaluation of JAK2 V617F mutation in chronic myeloproliferative disorders. PMID- 16270041 TI - Reply to Rumpold et al. PMID- 16270042 TI - The role of P-glycoprotein in imatinib resistance. PMID- 16270043 TI - Gene expression profiling of acute promyelocytic leukaemia identifies two subtypes mainly associated with flt3 mutational status. AB - Acute promyelocytic leukaemia (APL) is a well-defined disease characterized by a typical morphology of leukaemic cells, the presence of t(15;17) translocation and the unique sensitivity to the differentiating effect of all-trans retinoic acid. Nevertheless, some aspects are variable among APL patients, with differences substantially related to morphological variants, peripheral leukocytes count, the presence of a disseminated intravascular coagulopathy, different PML/RARalpha isoforms (long, variable or short) and Fms-like tyrosine kinase 3 (Flt3) mutations. In order to better define this variability, we investigated the gene expression profiles of 18 APL cases revealing, besides a high uniformity in gene expression pattern, the presence of few robust differences among patients able to identify, by an unsupervised analysis, two major clusters of patients characterized by different phenotypes (hypogranular M3v vs classical M3) and by the presence or absence of Flt3 internal tandem duplications (ITDs). Further supervised analysis confirmed that Flt3 status was the APL parameter best associated with these two subgroups. We identified, between Flt3 wild-type and Flt3-ITDs subsets, 147 differentially expressed genes that were involved in the cytoskeleton organization, in the cell adhesion and migration, in the proliferation and the coagulation/inflammation pathways as well as in differentiation and myeloid granules constitution suggesting a role of Flt3 mutations in the pathogenesis and clinical manifestations of APL. PMID- 16270044 TI - The NF-kappaB pathway blockade by the IKK inhibitor PS1145 can overcome imatinib resistance. AB - Imatinib represents at present the most attractive therapy for BCR-ABL positive leukemias, even though a percentage of CML patients develop resistance to this compound. For these resistant patients a therapeutic approach based on a combination of drugs is more likely to be effective. In the last years, constitutive NF-kappaB/Rel activity has been demonstrated in several hematological malignancies. As a result, NFkB/Rel-blocking approaches have been proposed as antineoplastic strategies. Furthermore, the identification of specific kinases within the NF-kappaB activation pathway offers a selective target to address tailored therapies. In the current study, we show that the IKK inhibitor PS1145 is able to inhibit the proliferation of CML cell lines and primary BM cells. Moreover, the addition of Imatinib increases the effects of PS1145 in resistant cell lines and BM cells from resistant patients, with a further increase of apoptosis and inhibition of proliferation and colony growth. Our data provide the rational for a new therapeutic approach, which combines Imatinib and the IKK inhibitor PS1145 in CML resistant patients. PMID- 16270045 TI - Accurate detection of asparagine synthetase (ASNS) using quantitative real-time PCR (qRT-PCR), without requiring DNaseI treatment. PMID- 16270046 TI - Differential expression of NF-kappaB pathway genes among peripheral T-cell lymphomas. AB - Nuclear factor kappa B (NF-kappaB) is one important pathway in T-cell proliferation and survival. In a previously reported microarray study, we found NF-kappaB pathway genes differentially expressed between peripheral (PTCL) and lymphoblastic lymphomas. Here, we investigated the expression of NF-kappaB pathway genes using cDNA microarrays in a group of 62 PTCL and in reactive lymph nodes. We found two different subgroups of PTCL based on the expression of NF kappaB pathway genes. One-third of PTCL showed clearly reduced expression of NF kappaB genes, while the other group was characterized by high expression of these genes. This distinction was found among all T-cell lymphoma categories analyzed (PTCL unspecified, angioimmunoblastic, cutaneous and natural killer/T lymphomas) with the exception of anaplastic lymphomas (ALCL), which were characterized by reduced NF-kappaB expression in anaplastic cells. Quantitative RT-PCR and immunohistochemical analysis of NF-kappaB-p65 protein confirmed these differences among PTCL subgroups. Importantly, we found that differentiation between NF kappaB-positive and -negative PTCL could be of clinical interest. The expression profile associated to reduced expression of NF-kappaB genes was significantly associated with shorter survival of patients and seems to be an independent prognostic factor in a multivariate analysis. PMID- 16270047 TI - A familiar face at the FDA. PMID- 16270049 TI - Mental health research emerges from hurricane ruin. PMID- 16270050 TI - Lack of leadership once again plagues US drug agency. PMID- 16270051 TI - In Bangladesh, arsenic-free water set to flow from cheap new filter. PMID- 16270052 TI - Poor records pinpoint lack of attention to gender gaps in science. PMID- 16270053 TI - Electrical impulses recharge spinal cord injury research. PMID- 16270054 TI - Spanish flu papers put spotlight on 'dual use' decisions. PMID- 16270055 TI - Neuroscience meeting draws fire over Dalai Lama lecture. PMID- 16270057 TI - Double jeopardy. PMID- 16270058 TI - Profile: Marie Filbin. PMID- 16270059 TI - NMR indicates that the small molecule RITA does not block p53-MDM2 binding in vitro. PMID- 16270061 TI - PrPSc in mammary glands of sheep affected by scrapie and mastitis. PMID- 16270062 TI - Angiotensin-converting enzyme as a GPIase: a critical reevaluation. PMID- 16270063 TI - Male fertility is dependent on dipeptidase activity of testis ACE. PMID- 16270065 TI - Ethics guidelines for research with the recently dead. AB - The objective of the multidisciplinary expert Consensus Panel on Research with the Recently Dead (CPRRD) was to craft ethics guidelines for research with the recently dead. The CPRRD recommends that research with the recently dead: (i) receive scientific and ethical review and oversight; (ii) involve the community of potential research subjects; (iii) be coordinated with organ procurement organizations; (iv) not conflict with organ donation or required autopsy; (v) use procedures respectful of the dead; (vi) be restricted to one procedure per day; (vii) preferably be authorized by first-person consent, though both general advance research directives and surrogate consent are acceptable; (viii) protect confidentiality; (ix) not impose costs on subjects' estates or next of kin and not involve payment; (x) clearly explain ultimate disposition of the body. PMID- 16270066 TI - Diuretic soothes seizures in newborns. PMID- 16270067 TI - Extinct 1918 virus comes alive. PMID- 16270068 TI - Cancer: the matrix is now in control. PMID- 16270070 TI - Neurons inflict self-harm. PMID- 16270071 TI - TLRs play good cop, bad cop in the lung. PMID- 16270072 TI - Vaccinating transplant recipients. PMID- 16270076 TI - Shifting paradigms in lung cancer. PMID- 16270073 TI - Protecting the host naturally. PMID- 16270077 TI - What is the value of screening for prostate cancer in the US? PMID- 16270078 TI - Does current evidence justify prostate cancer screening in Europe? PMID- 16270092 TI - What influence does hormone replacement therapy have on endometrial cancer risk? PMID- 16270093 TI - Is fractionated stereotactic radiotherapy effective in pituitary adenomas? PMID- 16270094 TI - Is radiotherapy optimally combined with chemotherapy in elderly patients with limited-stage small-cell lung cancer? PMID- 16270095 TI - Can adjuvant chemotherapy improve survival in patients with early-stage, resected non-small-cell lung cancer? PMID- 16270096 TI - EGFR inhibitors: what have we learned from the treatment of lung cancer? AB - Tyrosine kinase inhibitors directed against the epidermal growth factor receptor (EGFR) are the first molecular-targeted agents to be approved in the US and other countries for the treatment of advanced non-small-cell lung cancer after failure of chemotherapy. Some patient characteristics, such as never-smoking, female gender, East Asian origin, adenocarcinoma histology, and bronchioloalveolar subtype, are associated with a greater benefit from treatment with EGFR inhibitors. Recently, studies have identified gene mutations targeting the kinase domain of the EGFR that are related to the response to inhibitors. Most EGFR mutations predict a higher benefit from treatment compared with wild-type receptors and are correlated with clinical features related to better outcome; some EGFR mutations, however, confer drug resistance. The analysis of material usually available from lung cancer patients, using techniques such as direct sequencing to determine EGFR mutational status, can be technically challenging. In this regard, high EGFR copy number and EGFR protein detected by immunohistochemistry can also be used to select those patients who would benefit from treatment. Prospective validation of biological and clinical markers of sensitivity needs to be performed. PMID- 16270097 TI - Angiogenic inhibitors: a new therapeutic strategy in oncology. AB - Angiogenesis is a multistep, complex and tightly regulated process that is necessary for tumor growth and metastasis. Based on data of preclinical models, several antiangiogenic compounds has been shown to modify activated tumor endothelium, which suggests that these compounds can improve cytotoxic drug delivery. Such agents have entered clinical trials as single agents or in combination with cytotoxic drugs, and have shown promising antitumor activity. The pharmacodynamic and pharmacokinetic characteristics of antiangiogenic drugs are reviewed here. Most of the early clinical testing of these agents was conducted in patients with advanced disease resistant to standard therapies. Phase III trials compared the efficacy of standard chemotherapy alone with standard chemotherapy in combination with an experimental angiogenesis inhibitor. Although some of these studies were negative or controversial, recent studies validated in large clinical trials with an anti-vascular endothelial growth factor antibody demonstrated significant clinical benefit and renewed enthusiasm for this therapeutic strategy. This review describes the clinical studies of antiangiogenic agents and highlights the challenges related to choosing appropriate strategies for the selection of patients, study design and choice of appropriate endpoints for the studies' development. PMID- 16270099 TI - Recurrent endometrial stromal tumors with smooth-muscle differentiation and a protracted clinical course. AB - BACKGROUND: A 24-year-old white female gravida 1, para 0010, presented with heavy vaginal bleeding and abdominal cramps of 2 weeks' duration. Medical history was remarkable for spontaneous abortion 5 years previously. She had no significant family history or other gynecological problems. Physical examination revealed tissue fragments and blood clots oozing from the cervical os, and her uterus was diffusely enlarged. INVESTIGATIONS: Physical examination, ultrasound, uterine dilation and curettage, immunohistochemistry and human androgen receptor gene clonality analysis, uterine sonohistogram, MRI and exploratory laparotomy. DIAGNOSIS: Intrauterine dissemination of endometrial stromal tumors with smooth muscle differentiation. MANAGEMENT: Partial wedge resection of the anterior uterine wall via abdominal myomectomy and total abdominal hysterectomy. PMID- 16270098 TI - Drug Insight: Metastatic colorectal cancer--oral fluoropyrimidines and new perspectives in the adjuvant setting. AB - Capecitabine and uracil/ftorafur (UFT) are prodrugs of 5-fluorouracil (5-FU) that can be administered orally. Both drugs have been shown to be as effective as bolus 5-FU and folinic acid (FA), both as adjuvant treatment, and for the treatment of metastatic colorectal cancer. However, as the addition of oxaliplatin to 5-FU has been shown to improve disease-free survival in adjuvant therapy, oxaliplatin/5-FU is currently regarded as the standard adjuvant treatment, rather than fluoropyrimidine monotherapy. For the treatment of metastatic colorectal cancer, improved response rates and prolonged survival have been reported when irinotecan or oxaliplatin was added to 5-FU/FA; a further increase in efficacy was shown when bevacizumab, an antibody to vascular endothelial growth factor, was added to chemotherapy. Studies investigating the substitution of infusional 5-FU with oral compounds in combination therapy (e.g. FOLFOX versus capecitabine/oxaliplatin) are ongoing, but preliminary results in metastatic patients advise caution because of increased toxicity when used with irinotecan, and statistically non-significant trends of a decreased efficacy when used with oxaliplatin. Therefore, the combined use of oral 5-FU prodrugs plus irinotecan or oxaliplatin is not recommended at present. Oral 5-FU prodrugs, however, offer a convenient and less toxic treatment option in adjuvant treatment and in the treatment of advanced colorectal carcinoma in patients who do not want an intensified regimen, those that have contraindications for implanted venous access devices, and those who are not candidates for a combination therapy with irinotecan and oxaliplatin. PMID- 16270100 TI - The structure of the flock house virus B2 protein, a viral suppressor of RNA interference, shows a novel mode of double-stranded RNA recognition. AB - We report the structure of the flock house virus B2 protein, a potent suppressor of RNA interference (RNAi) in animals and plants. The B2 protein is a homodimer in solution and contains three alpha-helices per monomer. Chemical shift perturbation shows that an antiparallel arrangement of helices (alpha2/alpha2') forms an elongated binding interface with double-stranded RNA (dsRNA). This implies a novel mode of dsRNA recognition and provides insights into the mechanism of RNAi suppression by B2. PMID- 16270103 TI - The changing treatment paradigm in patients with newly diagnosed multiple myeloma: implications for nursing. AB - PURPOSE/OBJECTIVES: To review the changing treatment paradigm for newly diagnosed multiple myeloma and its implications for nursing. DATA SOURCES: Journal articles, textbooks, published research data. DATA SYNTHESIS: The treatment approaches to newly diagnosed multiple myeloma are varied, and no consensus exists about the best choice of induction therapy prior to high-dose chemotherapy with autologous stem cell transplantation. Novel therapies that have shown strong clinical activity in patients with relapsed or refractory myeloma currently are being explored as first-line therapy with associated higher incidence of serious complications. CONCLUSIONS: Novel approaches in the treatment of newly diagnosed multiple myeloma may lead to better overall patient survival. Research is ongoing to find ways to improve progression-free and overall survival in patients with multiple myeloma. IMPLICATIONS FOR NURSING: Oncology nurses play vital roles in the assessment and monitoring of serious complications associated with various therapies for patients with newly diagnosed multiple myeloma. Key responsibilities include safe and effective administration of complex chemotherapeutic regimens, management of side effects, patient and family education, and coordination of a multidisciplinary approach. PMID- 16270101 TI - The Caenorhabditis elegans homologue of the proto-oncogene ect-2 positively regulates RAS signalling during vulval development. AB - Guanine nucleotide exchange factors (GEFs) regulate the activity of small GTP binding proteins in a variety of biological processes. We have identified a gain of-function mutation in the Caenorhabditis elegans GEF ect-2, the homologue of the mammalian ect2 proto-oncogene that has an essential role during cytokinesis. Here, we report that, in addition to its known function during mitosis, ECT-2 promotes the specification of the primary vulval cell fate by activating RAS/mitogen-activated protein kinase (MAPK) signalling before the end of the S phase. Epistasis analysis indicates that ECT-2 crosstalks to the canonical RAS/MAPK cascade upstream of the RAS GEF SOS-1 by means of a RHO-1 signalling pathway. Our results raise the possibility that the transforming activity of the mammalian ect-2 oncogene could be due to hyperactivation of the RAS/MAPK pathway. PMID- 16270104 TI - Sleep wake disturbances in people with cancer and their caregivers: state of the science. AB - PURPOSE/OBJECTIVES: To review the state of the science on sleep/wake disturbances in people with cancer and their caregivers. DATA SOURCES: Published articles, books and book chapters, conference proceedings, and MEDLINE, the Cumulative Index to Nursing and Allied Health Literature, PsycINFO, and the Cochrane Library computerized databases. DATA SYNTHESIS: Scientists have initiated studies on the prevalence of sleep/wake disturbances and the etiology of sleep disturbances specific to cancer. Measurement has been limited by lack of clear definitions of sleep/wake variables, use of a variety of instruments, and inconsistent reporting of sleep parameters. Findings related to use of nonpharmacologic interventions were limited to 20 studies, and the quality of the evidence remains poor. Few pharmacologic approaches have been studied, and evidence for use of herbal and complementary supplements is almost nonexistent. CONCLUSIONS: Current knowledge indicates that sleep/wake disturbances are prevalent in cancer populations. Few instruments have been validated in this population. Nonpharmacologic interventions show positive outcomes, but design issues and small samples limit generalizability. Little is known regarding use of pharmacologic and herbal and complementary supplements and potential adverse outcomes or interactions with cancer therapies. IMPLICATIONS FOR NURSING: All patients and caregivers need initial and ongoing screening for sleep/wake disturbances. When disturbed sleep/wakefulness is evident, further assessment and treatment are warranted. Nursing educational programs should include content regarding healthy and disrupted sleep/wake patterns. Research on sleep/wake disturbances in people with cancer should have high priority. PMID- 16270106 TI - Approaching death: a phenomenologic study of five older adults with advanced cancer. AB - PURPOSE/OBJECTIVES: To explore the lived experience and the associated meaning of approaching death among older adults with advanced cancer. RESEARCH APPROACH: A phenomenologic inquiry. SETTING: Urban cancer center. PARTICIPANTS: 5 individuals diagnosed with advanced cancer who were 65 years or older. METHODOLOGIC APPROACH: A series of semistructured interviews were tape-recorded, transcribed verbatim, analyzed, and developed into narrative summations. FINDINGS: The study elucidated the experience of approaching death from advanced cancer. The insightful and compelling narratives of five individuals' experiences suggest that genuine caring, compassionate honesty from trusted healthcare professionals, cautious hopefulness maintained by patients and their loved ones, unquestioned faith, an involvement in desired life activities, and positive interactions within the healthcare system and in personal relationships were meaningful to participants. CONCLUSIONS: Knowledge developed from dying patients allows for an awareness that moves beyond assumption toward an in-depth understanding that can enable healthcare professionals to design meaningful care for these individuals. INTERPRETATION: Knowledge gained directly from patients' experiences of what is meaningful and helpful as death approaches can guide effective interventions to improve palliative care practices and enhance overall quality of life while living with an incurable illness. Nurses have the ability to contribute meaningfully to patients facing death by relating to them with genuine care that acknowledges their humanity, conveying information with a compassionate honesty that recognizes the impact it will hold for patients and their loved ones, supporting patients' faith and cautious hopes, assisting patients in enjoying an active involvement in life within their limitations, and providing a safe and trusting environment in which to receive care. PMID- 16270107 TI - Predictors of use of complementary and alternative therapies among patients with cancer. AB - PURPOSE/OBJECTIVES: To determine predictors of use of complementary and alternative medicine (CAM) therapies among patients with cancer. DESIGN: Secondary analysis of two federally funded panel studies. SETTING: Urban and rural communities in the midwestern United States. SAMPLE: Patients with lung, breast, colon, or prostate cancer (N = 968) were interviewed at two points in time. 97% received conventional cancer treatment, and 30% used CAM. The sample was divided evenly between men and women, who ranged in age from 28-98; the majority was older than 60. METHODS: Data from a patient self-administered questionnaire were used to determine CAM users. Responses indicated use of herbs and vitamins, spiritual healing, relaxation, massage, acupuncture, energy healing, hypnosis, therapeutic spas, lifestyle diets, audio or videotapes, medication wraps, and osteopathic, homeopathic, and chiropractic treatment. MAIN RESEARCH VARIABLES: Dependent variable for analysis was use or nonuse of any of the identified CAM therapies at time of interviews. Independent variables fell into the following categories: (a) predisposing (e.g., gender, age, race, education, marital status), (b) enabling (e.g., income, health insurance status, caregiver presence, geographic location), and (c) need (e.g., cancer stage, site, symptoms, treatment, perceived health need). FINDINGS: Significant predictors of CAM use were gender, marital status, cancer stage, cancer treatment, and number of severe symptoms experienced. CONCLUSIONS: Patients with cancer are using CAM while undergoing conventional cancer treatment. IMPLICATIONS FOR NURSING: Nurses need to assess for CAM use, advocate for protocols and guidelines for routine assessment, increase knowledge of CAM, and examine coordination of services between conventional medicine and CAM to maximize positive patient outcomes. PMID- 16270108 TI - Burnout in clinical research coordinators in the United States. AB - PURPOSE/OBJECTIVES: To assess burnout among clinical research coordinators (CRCs) and to determine which personal and job-related factors are associated with burnout. DESIGN: Random, stratified, cross-sectional mail survey. SETTING: CRCs from membership lists of clinical research organizations. SAMPLE: 252 CRCs in the United States. To be included in the study, participants must have been in their current position longer than six months and involved in clinical trial coordination or data management. Of 2,770 records, 900 CRCs were mailed questionnaires; 35% (316) responded, and 252 of those were eligible for analysis. Eligible respondents were Caucasian (86%), female (94%), and employed full-time (92%) in an oncology setting (71%). METHODS: Respondents completed mailed self administered questionnaires measuring burnout, job satisfaction, personality characteristics, perceived work overload, and selected personal- and employment related data. Data analyses included descriptive, univariate, and multivariate statistics. MAIN RESEARCH VARIABLE: Burnout. FINDINGS: About 70% of respondents were satisfied with their job, and 74% would still choose the clinical research profession. Approximately 44% reported high emotional exhaustion, a component of burnout. Factors independently associated with high emotional exhaustion burnout were low satisfaction with job (p 0.0001), high perceived daily workload (p 0.0001), and low endurance personality (p = 0.002). CONCLUSIONS: Burnout is prevalent in CRCs. Job dissatisfaction, perceived daily work overload, low endurance, and nurturance personality traits were associated with high burnout. IMPLICATIONS FOR NURSING: Nurses are involved significantly in clinical trial coordination. High burnout rates have potentially negative implications for data quality and productivity in clinical trial data management-important values for nursing and the clinical research profession. PMID- 16270109 TI - Creating the basis for a breast health program for female survivors of Hodgkin disease using a participatory research approach. AB - PURPOSE/OBJECTIVES: To learn from female survivors of Hodgkin disease about their perceptions of their current health status and future health risks, self-care practices to prevent or diminish health risks, and what kind of breast health program could benefit them, including the most effective methods and optimal times for learning about breast health. DESIGN: Participatory research using focus groups. SETTING: Urban pediatric cancer center. SAMPLE: 1 African American and 19 Caucasian female survivors of Hodgkin disease aged 16-26 years, diagnosed at least two years before the start of the study, and treated with mantle radiation therapy. Participants were recruited during visits to an outpatient clinic. METHODS: Six open-ended questions were asked during three separate focus group sessions. Transcribed data were evaluated by content analysis techniques and analyzed to identify common themes. MAIN RESEARCH VARIABLES: Current health status and perceived health risks, current health practices, and effective methods and timing for breast health teaching. FINDINGS: Survivors reported feeling damaged by their cancer and its treatment and perceived that they were at risk for breast cancer. Self-care and risky behaviors also were reported. Internal influences (e.g., fear) and external influences (e.g., family) motivated survivors to participate in health promotion activities. Effective methods identified for learning about breast health included having access to other survivors, being respected as an adult, and having one-on-one staff teaching and peer support. The preferred timing of teaching varied, but survivors generally supported a gradual provision of information. CONCLUSIONS: A positive listening environment is important for developing a breast health program for survivors. An essential first step is to create an opportunity for survivors to tell about their experiences with cancer, including its impact on their lives. Information regarding breast health must be provided in multiple formats during and after treatment if good practices are to be undertaken. IMPLICATIONS FOR NURSING: The provision of adequate information during and after therapy as well as peer counseling in a positive listening environment are important in helping survivors participate in health promotion activities. PMID- 16270110 TI - Perspectives on the impact of ovarian cancer: women's views of quality of life. AB - PURPOSE/OBJECTIVES: To describe quality-of-life (QOL) concerns particular to women with ovarian cancer and to examine whether subgroups of patients with ovarian cancer have significantly different QOL concerns. DESIGN: Mailed survey. SAMPLE: Readership of an ovarian cancer newsletter. METHODS: A total of 1,383 surveys were received in response to the survey's inclusion in the November 2002 issue of Conversations! The International Newsletter for Those Fighting Ovarian Cancer, a monthly newsletter circulated to 3,300 women with ovarian cancer (response rate = 42%). Women were asked to complete the 45-item City of Hope QOL Ovarian Cancer Tool (QOL-OVCA) and a short demographic questionnaire. MAIN RESEARCH VARIABLES: Patients' QOL-OVCA scores were compared across six independent variables, including disease status, age at diagnosis, stage at diagnosis, marital status, household income, and use of alternative therapy, controlling for survival time. FINDINGS: Ovarian cancer survivors reported significant QOL concerns across dimensions of physical, psychological, social, and spiritual well-being. CONCLUSIONS: Patients with ovarian cancer experience particular QOL concerns requiring support. Future research is needed to describe the needs of survivors in more diverse populations and to develop and test interventions that can address these QOL concerns. IMPLICATIONS FOR NURSING: Women with ovarian cancer experience QOL concerns common to other patients with cancer and some that are particular to ovarian cancer. Nurses should assess for and aggressively address these QOL concerns. PMID- 16270111 TI - Potential mechanisms for chemotherapy-induced impairments in cognitive function. AB - PURPOSE/OBJECTIVES: To review the domains of cognitive function and their corresponding neuroanatomic structures as well as present current evidence for neurotoxicity associated with specific chemotherapeutic agents and potential mechanisms for chemotherapy-induced cognitive impairments. DATA SOURCES: Published research articles, review articles, and textbooks. DATA SYNTHESIS: Chemotherapy does not appear to cross the blood-brain barrier when given in standard doses; however, many chemotherapy drugs have the potential to cause cognitive impairments through more than one mechanism. In addition, patient factors may be protective or place individuals at higher risk for cognitive impairments. CONCLUSIONS: Although evidence of chemotherapy-induced impairments in cognitive function exists, no clinical studies have attempted to elucidate the mechanisms for chemotherapy-induced impairments in cognitive function. In addition, further studies are needed to determine predictive factors, potential biomarkers, and relevant assessment parameters. IMPLICATIONS FOR NURSING: The ability to identify high-risk patients has important implications for practice in regard to informed consent, patient education about the effects of treatment, and preventive strategies. PMID- 16270112 TI - Cancer care from the perspectives of older women. AB - PURPOSE/OBJECTIVES: To understand how older age affects cancer care, from the perspectives of older women. RESEARCH APPROACH: Qualitative, participatory. SETTING: Urban southern region of Ontario, Canada. PARTICIPANTS: Purposive sample (age groups and income) of 15 women diagnosed with cancer at age 70 or older; 10 women were diagnosed with breast cancer, 5 with gynecologic cancer. METHODOLOGIC APPROACH: Two face-to-face interviews, with data analysis in collaboration with the project team based on constructivist grounded theory, including negative case analysis. MAIN RESEARCH VARIABLES: Age, experience of cancer care. FINDINGS: Age related life and health circumstances intersect with professional practice and wider social contexts and are implicated in treatment decision making, including decisions against treatment, as well as in the day-to-day "getting around" that cancer care requires. CONCLUSIONS: The nursing history should be holistic in scope, attending to the supportive care domains to elicit older women's physical, social, practical, informational, psychological, and spiritual needs after a diagnosis of cancer. History taking should draw forward older women's life contexts and examine these contexts in relation to cancer care, including treatment decision making. INTERPRETATION: Individual-level care and systems advocacy are required to ensure that older women's worries about sustaining independence, including worries generated by inadequacies in home-based care, do not act as determinants of treatment choices. PMID- 16270113 TI - Patient communication following head and neck cancer surgery: a pilot study using electronic speech-generating devices. AB - PURPOSE/OBJECTIVES: To describe the communication of patients who received electronic speech-generating devices (SGDs) following surgical procedures for head or neck cancer. DESIGN: Exploratory, complementary mixed methods. SETTING: Otolaryngology surgical inpatient unit of an urban teaching hospital. SAMPLE: 10 purposively selected patients with a mean age of 57.1 years (SD = 12.8 years) and moderately severe illness (Acute Physiology and Chronic Health Evaluation III score mean = 27.1 + 13.2) who had SGDs in their hospital rooms for 9.1 + 6.2 days. METHODS: Observation, interviews, questionnaires, and clinical record review. MAIN RESEARCH VARIABLES: Communication methods, communication content, SGD use, communication quality (i.e., ease and user satisfaction), barriers to SGD use, and patient clinical characteristics. FINDINGS: SGDs were used in message construction in 8 (17%) out of 48 total observed communication events. Writing (31%) and nonverbal communication (46%) were the most frequently observed primary methods of communication used by patients with head and neck cancer postoperatively. Five patients demonstrated occasional SGD use with or without cuing, and one used the SGD as the dominant communication method. Ease of Communication Scale scores showed only slightly less difficulty with communication when compared to a historic control group. Patients initiated communications more often when SGDs were used in message construction. Poor device positioning, staff unfamiliarity with SGDs, and patient preference and ability for writing were barriers to SGD use. CONCLUSIONS: Although writing and making gestures were the most common communication methods, SGDs were used successfully by selected patients and may be particularly beneficial for constructing complex messages during conversation. IMPLICATIONS FOR NURSING: SGDs may be an appropriate assistive communication strategy for postoperative patients with head and neck cancer. Nurses can facilitate effective patient communication with SGDs by cuing patients on device options and positioning SGDs within easy reach. PMID- 16270115 TI - The standardization of radiation skin care in British Columbia: a collaborative approach. AB - PURPOSE/OBJECTIVES: To develop evidence-based practice guidelines for and standardize the care of radiation skin reactions. DATA SOURCES: Peer-reviewed scientific journals and texts and a survey of the guidelines in use at leading cancer treatment facilities in Canada, the United States, the United Kingdom, and Australia. DATA SYNTHESIS: A formal reference document with recommended guidelines was developed. Consensus was obtained from all relevant disciplines, and the guidelines were implemented successfully into practice. CONCLUSIONS: The document introduced a major change in practice from the maintenance of a dry radiation treatment area to the promotion of skin cleanliness and hydration, as well as the adoption of the principles of moist wound healing. Annual review indicated that dissemination of (94%) and compliance with (78%) the guidelines were good. IMPLICATIONS FOR NURSING: The process to develop, obtain consensus for, and implement evidence-based practice guidelines was an exemplary demonstration of teamwork and interdisciplinary collaboration. PMID- 16270114 TI - A cognitive behavioral intervention for symptom management in patients with advanced cancer. AB - PURPOSE/OBJECTIVES: To evaluate the effectiveness of a cognitive behavioral intervention in decreasing symptom severity in patients with advanced cancer undergoing chemotherapy. DESIGN: Prospective, randomized clinical trial based on cognitive behavioral theory. SETTING: Six urban cancer centers in the midwestern United States. SAMPLE: 124 patients 21 years of age or older were recruited and randomized to receive conventional care or conventional care and an intervention. Participants were newly diagnosed with stage III, stage IV, or recurrent cancer (solid tumor or non-Hodgkin lymphoma), undergoing chemotherapy, cognitively intact, and able to read and speak English. METHODS: Data were gathered via telephone interviews at baseline and 10 and 20 weeks after randomization. Nurses with experience in oncology delivered a five-contact, eight-week intervention aimed at teaching patients problem-solving techniques to affect symptom severity. MAIN RESEARCH VARIABLES: Gender, site of cancer, age, symptom severity and depressive symptoms at baseline, group (i.e., experimental versus control), and total symptom severity. FINDINGS: Patients in the experimental group and those with lower symptom severity at baseline had significantly lower symptom severity at 10 and 20 weeks; the experimental difference at 20 weeks occurred primarily in those 60 years of age and younger. Depressive symptoms at baseline predicted symptom severity at 20 weeks; however, age, gender, and site of cancer did not affect symptom severity at either time point. CONCLUSIONS: A cognitive behavioral intervention to teach problem-solving skills can be effective for patient symptom self-management during and following an intervention. IMPLICATIONS FOR NURSING: Problem-solving strategies should be included in educational programs for patients with advanced cancer, particularly those 60 years of age and younger. PMID- 16270116 TI - Collaborative breast health intervention for African American women of lower socioeconomic status. AB - PURPOSE/OBJECTIVES: To describe all phases of a collaborative breast health intervention delivered by paraprofessionals or specially trained community health advisors (CHAs) for African American women designed to increase mammography screening. DESIGN: Collaborative pretest, post-test breast health intervention. SETTING: Large city in Ohio. SAMPLE: 68 African American women with a median age of 57.8 (SD = 5.28) obtained mammography screening and participated in the breast health intervention. METHODS: Specially trained CHAs used aggressive recruitment strategies to increase mammography screening and knowledge of breast health and mammography screening in African American women aged 50 and older. MAIN RESEARCH VARIABLES: Knowledge scores of breast health and mammography screening. FINDINGS: Ninety women (81%) met the inclusion criteria and were recruited into the intervention, but only 68 (76%) obtained mammography screening. The women demonstrated increased knowledge by change in pre- to post-test scores. Several questions were statistically significant. CONCLUSIONS: Collaborative breast health interventions delivered by trained CHAs are effective in increasing screenings as well as knowledge of breast health and mammography screening in African American women. The unique role of the CHA is especially important in recruitment of hard-to-reach women and was vital to the success of the educational intervention. Most importantly, the women valued the individualized attention to their breast health and agreed to share the information with significant others. Further collaborative interventions designed to increase screenings and increase knowledge of breast health and mammography screening are needed to reduce the health disparities of later-stage detection and poorer survival of breast cancer in African American women. IMPLICATIONS FOR NURSING: Oncology nurses should build on the findings and deliver further outreach programs to increase mammography screening and knowledge of breast health in a larger number of women of lower socioeconomic status. Future research is needed to determine the influence of reminder phone calls for mammography screening. Oncology nurses should incorporate evaluation strategies at baseline and periodically throughout an intervention to provide more comprehensive data and enhance the credibility of findings. To maximize success, oncology nurses should work collaboratively with other healthcare professionals such as certified x-ray technicians and influential people in the community to increase knowledge of breast health and mammography screening. PMID- 16270117 TI - Hepatitis C and hemodialysis: a review. AB - Hepatitis C is a serious public health problem throughout the world; chronic renal patients are highly exposed to this infection. This could be due to a failure to identify carriers of this disease or because of a lack of truly effective biosafety measures implemented in the dialysis units. Molecular biology techniques have allowed for the understanding of this virus in detail, including its replication mechanisms. Epidemiological studies have been made throughout the world, with the goal of determining the dissemination dynamics of this agent, in addition to examining the predominance of the different genotypes, and the possible mutants that are involved. Many questions must still be answered concerning infection by Hepatitis C virus (HCV); this is especially important for immunosuppressed patients. PMID- 16270118 TI - Colonization by Streptococcus agalactiae during pregnancy: maternal and perinatal prognosis. AB - We reviewed colonization by group B Streptococcus beta-haemolyticus of Lancefield (SGB), or Streptococcus agalactiae, in pregnant women, and the consequences of infection for the mother and newborn infant, including factors that influence the risk for anogenital colonization by SGB. We also examined the methods for diagnosis and prophylaxis of SGB to prevent early-onset invasive neonatal bacterial disease. At present, it is justifiable to adopt anal and vaginal SGB culture as part of differentiated obstetrical care in order to reduce early neonatal infection. The rates, risk factors of maternal and neonatal SGB colonization, as well as the incidence of neonatal disease, may vary in different communities and need to be thoroughly evaluated in each country to allow the most appropriate preventive strategy to be selected. PMID- 16270119 TI - Antibiotic prophylaxis in orthopedic surgeries: the results of an implemented protocol. AB - Though the basic principles of antibiotic prophylaxis have been well established, there is still considerable incorrect usage, including how much is prescribed and especially in the duration of treatment, which is generally superior to what is indicated. The adequate use of these drugs contributes towards decreasing the time of internment of the patient, prevents surgical site infection (SSI), decreasing the development of resistant microorganisms, and towards reduced costs for the hospital pharmacy. A protocol for the use of antibiotic prophylaxis in the Orthopedics and Traumatology Service of the Hospital do Servidor Publico Estadual de Sao Paulo was developed. The objectives of the study were to promote rational antibiotic surgical prophylaxis, through the implantation of a protocol for the use of these drugs in a surgical unit, with the direct contribution of a druggist in collaboration with the Infection Control Committee, to evaluate the adhesion of the health team to the protocol during three distinct periods (daily pre-protocol, early post-protocol and late post-protocol) and to define the consumption of antimicrobials used, measured as daily defined dose. PMID- 16270120 TI - Use of antibiotics without medical prescription. AB - The inappropriate use of antibiotics for the treatment of infections is a worldwide problem that has implications for the cost of treatment and the development of resistant strains of bacteria. The use of antibiotics should follow specific criteria; they are on top of the list of self-medication drugs in countries that do not control their commercialization. OBJECTIVES: To determine the percentage of pharmacies that attend the public and sell antibiotics without medical prescription in a medium-sized city in Brazil, and analyze the variables involved in this procedure. MATERIALS AND METHODS: 107 of the 136 pharmacies registered in our city were evaluated. These pharmacies were visited by actresses who simulated having a sister with symptoms of a non-complicated rhino-sinusitis, so that they could obtain antibiotics without a medical prescription. Each pharmacy was visited only once; the only variable in the simulated clinical setting was the report of fever temperature, which was randomly assigned between 38.5 and 40 degrees Celsius. RESULTS: Antibiotics were offered in 58% of the pharmacies, and this offer was increased to 74% after the actresses insisted on having them. In 65.4% of the pharmacies, the actresses were attended by a pharmacist, and 84.2% of them said they would sell antibiotics. When the request for antibiotics was denied (26%), only 7.5% was due to absence of prescription. The most frequent reason for refusal to sell antibiotics, was because the attendant deemed it unnecessary (46.6%) CONCLUSION: Antibiotics can be easily bought in the great majority of the pharmacies in our town without a medical prescription and a clear indication. Fever temperature did not modify the attendant's indication of the drug. PMID- 16270121 TI - Intestinal microbiota of patients with bacterial infection of the respiratory tract treated with amoxicillin. AB - The intestinal tract harbors a huge diversity of metabolically-active aerobic and anaerobic bacteria that interact, forming a complex ecosystem. This microbiota has an important role in human metabolism, nutrition, immunity, and protection against colonization by pathogenic microorganisms. Several factors can influence the intestinal microbiota; these include age, diet, inflammatory and infectious processes, and the use of antimicrobials. We investigated the influence of bacterial infection of the respiratory tract and of amoxicillin therapy on the normal intestinal microbiota of patients. Bacterial infectious processes affecting the respiratory tract were found to influence the intestinal microbiota, significantly decreasing the number of colony-forming units (CFUs) of Bacteroides spp. and Lactobacillus spp. per gram of feces. The use of amoxicillin also influenced the intestinal microbiota, significantly decreasing the CFU of Bifidobacterium spp. and Lactobacillus spp./g of feces. Changes in the composition of the intestinal microbiota need to be observed, since a decrease in the normal microorganisms can pose a number of hazards for hosts, including decreased resistance to colonization. With proper follow-up, health-care teams can minimize such hazards by implementing suitable therapy- and diet-related measures, thus reducing the occurrence of detrimental effects on the gastrointestinal ecosystem. PMID- 16270122 TI - An outbreak of Acinetobacter baumannii septicemia in a neonatal intensive care unit of a university hospital in Brazil. AB - We studied an outbreak of two multi-drug resistant clones of Acinetobacter baumannii in the Neonatal Intensive Care Unit of the Uberlandia Federal University Hospital in Minas Gerais state, Brazil, and we analyzed the contribution of cross-transmission in the rise in infection rates. Eleven neonates who developed multi-drug resistant A. baumannii nosocomial infection were matched to 22 neonates who were admitted to the same unit and did not develop an infection during the outbreak period, in order to identify risk factors for infection. Three out of the 11 neonates died. Epidemiological investigation included molecular typing, using pulsed field gel electrophoresis. Prior to the outbreak, from December 2001 to March 2002, no case of infection by this microorganism was diagnosed. Environmental and healthcare worker hand cultures were negative. Nine isolates had similar pulsed field gel electrophoresis patterns and two had another clone. The first clone was brought into the unit by an infected patient who was transferred from another hospital without a history of antibiotic use. The second clone did have its origin clearly defined. Both infected groups led us to conclude that several factors contributed to infection with A. baumannii. These factors were: exposure to antibiotics and invasive devices, birth weight < 1500 g, age < or = 7 days and duration of hospitalization > or = 7 days. Based on logistic regression, infected neonates were more exposed to carbapenem and mechanical ventilation than the control group. Cross transmission between infants contributed to the rise in the rates of multi-drug resistant A. baumannii infection. PMID- 16270123 TI - Surgical site infection in a university hospital in northeast Brazil. AB - We examined prevention of surgical site infection (SSI) in a tertiary teaching hospital in northeast Brazil, from January 1994 to December 2003. The survey included 5,742 patients subjected to thoracic, urologic, vascular and general surgery. The criteria for diagnosing SSI were those of the Centers for Disease Control, USA, and the variables of the National Nosocomial Infection Surveillance risk index were used. Data analysis revealed that anesthetic risk scores, wound class and duration of surgery were significantly associated with SSI. A total of 296 SSIs were detected among the 5,742 patients (5.1%). The overall incidence of SSI was 8.8% in 1994; it decreased to 3.3% in 2003. In conclusion, the use of educational strategies, based on guidelines for SSI prevention reduced SSI incidence. Appropriate management of preoperative, intraoperative, and postoperative incision care, and a surveillance system based on international criteria, were useful in reducing SSI rates in our hospital. PMID- 16270125 TI - HIV-1 genotypes related to failure of nelfinavir as the first protease inhibitor treatment. AB - Combined antiretroviral therapy results in sustained viral suppression and a decrease in mortality and morbidity due to HIV infection. Intrinsic strength, durability and absence of cross-resistance are key factors in the selection of antiretrovirals. Failure with nelfinavir has been associated with two protease gene mutations, D30N and L90M. The D30N mutation does not result in cross resistance with other protease inhibitors, and it decreases viral fitness. In order to check for this mutation after failure with nelfinavir, the 246 HIV-1 genotyping test was performed on virus samples from 55 patients with failure of nelfinavir as the first protease inhibitor. Most (84%) of the viral strains were of subtype B. Nucleosides associated with mutations (NAM) were observed in 80% of the tests; no INS69, complex 151, K65R and L74V mutations, which give multi resistance to nucleoside analogue reverse transcriptase inhibitors to tenofovir and DDI, respectively, were observed. In the tests for protease gene mutations, the D30N mutation was found in 57%, L90M in 18% and the wild-type virus in 25%. These data are similar to published reports, showing that alternative therapies used after failure with nelfinavir may be more successful, as the D30N mutation does not cause cross-resistance to other protease inhibitors. PMID- 16270126 TI - A non-randomized comparative study using different doses of acyclovir to prevent herpes simplex reactivation in patients submitted to autologous stem cell transplantation. AB - The reactivation of Herpes Simplex virus (HSV) occurs in 70% to 80% of patients submitted to autologous stem cell transplantation (ASCT); it increases the severity of chemotherapy-induced mucositis. Therefore, the use of acyclovir in ASCT patients is considered standard practice. However, the minimum dose needed to prevent reactivation is a matter of debate. We compared two doses of acyclovir in a non-randomized fashion in 59 patients submitted to ASCT: 32 patients received a dose of 125 mg/m(2) IV every six hours and the subsequent 27 patients received a dose of 60 mg/m(2) IV every six hours. Viral excretion was evaluated through weekly viral culture of oral swabs. Grade 4 mucositis was more frequent in Group 1 (p= 0.03). The reactivation rates in Groups 1 and 2 were 9% and 4%, respectively (p= 0.62, 95% confidence interval -7 - 18). Prophylaxis with reduced doses of intravenous acyclovir seems to be as effective as a higher dose in inhibiting HSV reactivation, with a significant reduction in cost. Prospective randomized studies are needed to confirm our conclusions. PMID- 16270124 TI - The beta-chemokines MIP-1alpha and RANTES and lipoprotein metabolism in HIV infected Brazilian patients. AB - HIV patients are predisposed to the development of hypertriglyceridemia and hypercholesterolemia as a result of both viral infection and HIV infection therapy, especially the protease inhibitors. Chemokines and cytokines are present at sites of inflammation and can influence the nature of the inflammatory response in atherosclerosis. We investigated the correlation between biochemical variables and beta-chemokines (MIP-1alpha and RANTES) and the apolipoprotein E genotype in HIV-infected individuals. The apolipoproteins were measured by nephelometry. Triglycerides and total cholesterol were determined by standard enzymatic procedures. The beta-chemokines were detected by ELISA. The genetic category of CCR5 and apolipoprotein E were determined by PCR amplification and restriction enzymes. Immunological and virological profiles were assessed by TCD(4)+ and TCD(8)+ lymphocyte counts and viral load quantification. Positive correlations were found between apo E and CD(8)+ (p = 0.035), apo E and viral load (p = 0.018), MIP-1alpha and triglycerides (p = 0.039) and MIP-1a and VLDL (p = 0.040). Negative correlations were found between viral load and CD(4)+ (p = 0.05) and RANTES and CD(4)+ (p = 0.029). The beta-chemokine levels may influence lipid metabolism in HIV-infected individuals. PMID- 16270127 TI - Weil's disease: an unusually fulminant presentation characterized by pulmonary hemorrhage and shock. AB - A case of fulminant leptospirosis is presented, manifesting as rapid progression from acute undifferentiated febrile illness to refractory shock, jaundice, renal failure and massive pulmonary hemorrhage. The patient received aggressive intensive care unit support including prolonged intubation and ventilation. This case emphasizes that acute leptospirosis may well not be characterized by the classic scenario of a biphasic illness, but rather by a fulminant, monophasic illness. PMID- 16270128 TI - Pathologic study of a fatal case of dengue-3 virus infection in Rio de Janeiro, Brazil. AB - Dengue hemorrhagic fever (DHF) is a severe febrile disease, characterized by abnormalities in hemostasis and increased vascular permeability, which in some cases results in hypovolemic shock syndrome and in dengue shock syndrome. The clinical features of DHF include plasma leakage, bleeding tendency and liver involvement. We studied the histopathological features of a fatal case of dengue 3 virus infection. The patient, a 63-year old male, presented with an acute onset of severe headache, myalgia and maculopapular rash. Tissue fragments (liver, spleen, lung, heart, kidney and lymph nodes) were collected for light microscopy studies and stained by standard methods. Histopathology revealed severe tissue damage, caused by intense hemorrhage, interstitial edema and inflammation. Some tissue sections were also processed with the immunoperoxidase reaction, which revealed the dengue viral antigen. Dengue-3 virus was isolated and identified with electron microscopy in a C6/36 cell culture inoculated with the patient's serum. Viral particles were detected in the infected cell culture. PMID- 16270129 TI - [Good, pretty and cheap]. PMID- 16270130 TI - [What is neurologic monitoring at the bedside?]. PMID- 16270131 TI - [Should videolaparoscopy be routine in inguinal hernia?]. PMID- 16270132 TI - [Could the way of approaching the implantation of silicone breast prostheses alter the investigation of the sentinel lymph node in eventual cancer of the breast?]. PMID- 16270133 TI - [Non-invasive ventilation: when to use?]. PMID- 16270134 TI - [Marijuana Abuse]. PMID- 16270136 TI - [What is the importance of diagnostic video-assisted hysteroscopy in the climacteric?]. PMID- 16270135 TI - [Images in medicine. Multiple brain abscess ]. PMID- 16270141 TI - [Cardiac autonomic responses to head-up tilt in obese adolescents]. AB - INTRODUCTION: Obesity is associated with alterations of the autonomic nervous system. Previous studies in adults have shown that obese individuals present a decreased cardiac autonomic response to postural challenges. However, little is known about the impact of overweight on autonomic responses to passive postural stress in adolescents. OBJECTIVE: to compare cardiac autonomic responses to the head-up tilt maneuver between obese and non-obese adolescents by analyzing heart rate variability. METHODS: Fourteen obese adolescents (15.5+/-1.6 years) were compared with twenty non-obese subjects (15.4+/-0.8 years). Cardiac autonomic modulation was studied by heart rate variability (HRV) analysis of 5 minute RR interval recordings before and after a 70 masculine head-up tilt maneuver. HRV was analyzed according to the time domain (TD) and frequency-domain (FD) methods. The power spectral components were studied at low (LF) and high (HF) frequencies and as the LF/HF ratio. RESULTS: Obese adolescents demonstrated significantly lower HF normalized units (38.2+/-11.1 vs 53.9+/-15.5, p<0.05) and higher LF normalized units (60.7+/-11.3 vs 44.6+/-15.7, p<0.05) in the supine position. No difference was found in any HRV parameters after head-up tilt. When comparing differences between the orthostatic and supine positions, obese adolescents showed lesser changes of LF normalized units (22.4+/-12.6 vs 38+/-16.4, p<0.05) and HF normalized units (-21.9+/-12.4 vs -37.3+/-16.3, p<0.05). CONCLUSIONS: Obese adolescents have an abnormal cardiac autonomic response to the head-up tilt maneuver, characterized principally by a decreased parasympathetic response. PMID- 16270142 TI - [Predictive factors of rebleeding in cirrhotic patients submitted to Warren's surgery]. AB - BACKGROUND: Establish the predictive factors of rebleeding in cirrhotic patients submitted to the Warren's surgery. METHODS: 57 cirrhotic patients with good hepatic functional reserve and previous variceal bleeding that had not responded to clinical, endoscopic treatment were submitted to the Warren's surgery (distal splenorenal shunt). They were divided in two groups: I (31 had presented rebleeding at postoperative care) and II (26 had not presented new bleeding). Group I was again divided into 2 groups according to time of rebleeding: Group I.A (12 that presented rebleeding until hospital discharge) and Group I.B (19 that presented rebleeding after hospital discharge). We analyzed the clinical, endoscopic, laboratorial and Doppler-ultrasound information at the pre- and postoperative moments and also intrasurgery data that were compared between the established groups. RESULTS: Serum albumin and bilirubins were 3.33 mg% and 1.7 mg% in group I, 3.56 mg% and 1.16 mg% in group II. Portal flow and diameter were 0.24 cm/s and 1.34 cm in group I, and 0.18 cm/s and 1.21 cm in group II, respectively. In group I.A the volume of crystalloid given during surgery was 3.692 ml against 2.166 ml in group I.B. CONCLUSION: Albumin and total bilirubins dosages in the preoperative period, added to the value of the flow and portal diameter were predictive factors for rebleeding in cirrhotic patients submitted to the Warren's surgery. The volume of crystalloid given during the surgery was a predictive factor for early rebleeding. PMID- 16270143 TI - [Lipid profile and alteration of body composition in growth hormone deficient adults]. AB - BACKGROUND: The growth hormone deficiency (GHD) syndrome in adults and the increased associated cardiovascular risk have been extensively studied in recent years. Abnormal body composition with excess of visceral adiposity and adverse lipid profile are important features of this syndrome. Abnormal lipid profile has been described with increased levels of total cholesterol (C), LDL-cholesterol (LDL-C), triglycerides, decreased levels of HDL-cholesterol (HDL-C) and apolipoproteins abnormalities. METHODS: Lipid profile and the amount of visceral adipose tissue were studied in 31 GHD adults compared with a control group of healthy subjects matched for age, gender and body mass index (BMI). Visceral adipose tissue was evaluated by abdominal computed tomography and anthropometric measurements--BMI (kg/m2) and waist circumference (cm). The lipid profile was studied by measurement of C, LDL-C, HDL-C, triglycerides, apolipoproteins A and B, and Lipoprotein (a). RESULTS: The GHD adults showed increased visceral adipose tissue (156.66 +/- 72.72 vs. 113.51 +/- 32.97 cm2, p = 0.049), higher levels of triglycerides (158.58 +/- 80.29 vs. 97.17 +/- 12.37 mg/dl; p = 0.007) and lower HDL- cholesterol (45.41 +/- 13.30 vs. 55.34 +/- 14.31 mg/dl; p = 0.002). There were no differences in others aspects of lipid profile and anthropometric measurements. CONCLUSION: Growth Hormone Deficient adults showed increased visceral adipose tissue, higher levels of triglycerides and lower HDL- cholesterol levels. PMID- 16270144 TI - [The significance of the removal of the uterus for women: a qualitative study]. AB - OBJECTIVE: To identify the myths accepted by women about to undergo hysterectomy regarding the consequences that removal of their uterus might have on their lives. METHODS: A sample of 10 hospitalized women with surgical indication for hysterectomy was used to perform a qualitative study, with semi-structured interviews carried out during the preoperative period. A profile questionnaire was used with questions about personal features and semi-structured inquiries to facilitate deeper questioning. With authorization of the subjects, interviews were recorded and transcribed to perform a thematic analysis of the content. RESULTS: The myths most often mentioned during the interviews were: loss of femininity, frigidity, change in personality, possible change in body appearance (since they would be "hollow", "empty", "have a hole inside their belly"), interference in the affective and sexual life and the belief that their companion might change in relation to them (assuming that they would be hollow, cold, and with no sexual attraction). CONCLUSION: The interviewed women accepted a number of myths about the consequences that removal of their uterus would have on their lives. These myths are related to the social identity of the gender and to a lack of knowledge about their body its organs and functions. PMID- 16270145 TI - [Acute renal failure related to intravenous acyclovir]. AB - OBJECTIVE: The aim of this study was to evaluate the incidence and outcome of acute renal failure (ARF) in patients submitted to intravenous (IV) acyclovir treatment. METHODS: All patients over 13 years of age that used intravenous acyclovir for 5 or more days were retrospectively analyzed. When serum creatinine levels, previously in the normal range, increased above 2 mg/dl, the case was considered an ARF. Treatment and outcome of patients that developed ARF were analyzed. RESULTS: Eighty-five patients received IV acyclovir during the study period. Forty-one patients were included in the study. ARF developed in 8 out of 41 patients (19.5%). In the ARF cases, after beginning of treatment, the average time for increase of the serum creatinine levels was 4.2 days. Creatinine levels reached their peak in a mean time of 7.1 days (ranging from 3 to 14 days). Recovery of the renal function, evaluated by decrease of the creatinine level, varied from 1 to 7 days (mean of 3.6 days). ARF had a good outcome with hydration, lengthening of drug infusion time and adjustment of the drug dosage. CONCLUSION: Acyclovir induced ARF in 19.5% of the patients. All patients had a positive response with return to a normal renal function after hydration, lengthening of drug infusion time and adjustment of the drug dosage. None of the patients required treatment with hemodialysis. Acyclovir is a safe drug when administered with certain preventive measures. PMID- 16270146 TI - [Teleophthalmology as an auxiliary approach for the diagnosis of infectious and inflammatory ocular diseases: evaluation of an asynchronous method of consultation]. AB - PURPOSE: The purpose of this study was to evaluate the efficacy of teleophthalmology as an auxiliary approach for diagnosis of infectious and inflammatory ocular diseases and to study the main problems associated with the conclusion reached by consultation. METHODS: Two hundred and thirty three 35 mm slides (retinography or biomicroscopy photos) were digitalized and stored in a Compact Disc to be later analyzed in a computer monitor by two consultants, specialists in uveitis. The consultants filled out a form and a questionnaire, indicating the diagnosis and probable treatment for the analyzed cases also the main difficulties found to reach a conclusion of the consultation. The Kappa statistic, an assessment of the change-corrected agreement (reproducibility) among consultants, was calculated. Kappa values of > 0.7 indicated excellent agreement; values between 0.4 and 0.7 indicated fair to good agreement and values of < 0.4 indicated poor agreement. RESULTS: Agreement between the consultants and the final diagnosis was of 73.5%. Treatment suggestion or management was not possible in 8% of the cases for consultant A and in 10.4% for consultant B. The higher agreement rate among consultants, relating to the reasons that rendered diagnosis difficult, was found in relation to "lack of clinical data" and "low quality of images". CONCLUSION: Teleophthalmology, through the asynchronous method, was an efficient means for diagnosis of patients with infectious and inflammatory ocular diseases. The main problems affecting the teleophthalmology consultation were lack of detailed clinical data and low quality of the images. PMID- 16270147 TI - [Assessment of periodontal condition of kidney patients in hemodialysis]. AB - OBJECTIVES AND METHODS: Thirty patients undergoing hemodialysis were evaluated for their periodontal condition through plaque, calculus and gingival indexes; PSR and IgA present in the saliva were also appraised in order to determine the periodontal condition of patients presenting chronic kidney disease. RESULTS: Results obtained demonstrated that in the studied group the plaque, calculus and gingival indexes were high. With regard to these patients' requirement to undergo periodontal treatment, most patients submitted to hemodialysis needed better oral hygiene, in addition to supra and sub gingival scraping and topic application of fluor (code 2). As to the IgA amount present in the saliva, only three patients showed a low density. CONCLUSIONS: Patients presenting chronic kidney disease disclosed a tendency for greater bacterial plaque concentration, high formation of dental calculus suggesting the need for periodontal treatment comprising supra and sub gingival scraping. PMID- 16270148 TI - [Epidemiology of facial trauma]. AB - OBJECTIVES: This study aims to determine the incidence, etiology, severity of facial trauma and associated injuries enabling a greater understanding of its range and magnitude METHODS: A hundred and sixty four patients were selected with some degree of facial trauma regardless of gender, age and skin color. Data were analyzed by the Pearson chi2 statistical method. RESULTS: A male predominance was observed (78%) and its peak age was between 20 and 39 years. The major cause was interpersonal violence (48.1%), followed by fall (26.2%), run overs 6.4%), sports (5.4%), car accidents (4.2%), motorcycle accidents (3.1%), non-fall impacts (2.4%), occupational injuries (1.8%), gunshot wounds (1.2%), unspecific (1.2%). Contusion is the most common injury (23.8%), followed by fractures of the mandible (21.9%), Le Fort/pan facial/complex (17.8%), nasal bones (11.6%), zygoma (10.3%), tooth (9.1%), orbit (4.9%) and maxilla (0.6%). Associated injuries occurred mostly in run overs, but also because of car accident, fall and interpersonal violence. CONCLUSIONS: The causes of facial trauma are directly related to the age and type of lesion. No evidence was found that the causes were related to gender or severity of the lesion. PMID- 16270150 TI - Caspase family proteases and apoptosis. AB - Apoptosis, or programmed cell death, is an essential physiological process that plays a critical role in development and tissue homeostasis. The progress of apoptosis is regulated in an orderly way by a series of signal cascades under certain circumstances. The caspase-cascade system plays vital roles in the induction, transduction and amplification of intracellular apoptotic signals. Caspases, closely associated with apoptosis, are aspartate-specific cysteine proteases and members of the interleukin-1beta-converting enzyme family. The activation and function of caspases, involved in the delicate caspase-cascade system, are regulated by various kinds of molecules, such as the inhibitor of apoptosis protein, Bcl-2 family proteins, calpain, and Ca2+. Based on the latest research, the members of the caspase family, caspase-cascade system and caspase regulating molecules involved in apoptosis are reviewed. PMID- 16270151 TI - Cloning and alternative splicing analysis of Bombyx mori transformer-2 gene using silkworm EST database. AB - We have identified Bombyx mori transformer-2 gene (Bmtra-2) cDNA by blasting the EST database of B. mori. It was expressed in the whole life of the male and female silkworm and was observed as a band of 1.3 kb by Northern blot analysis. By comparing corresponding ESTs to the Bmtra-2 DNA sequence, it was revealed that there were eight exons and seven introns, and all splice sites of exons/introns conformed to the GT/AG rule. Bmtra-2 pre-mRNA can produce multiple mRNAs encoding six distinct isoforms of BmTRA-2 protein using an alternative splicing pathway during processing. Six types of Bmtra-2 cDNA clones were identified by reverse transcription-polymerase chain reaction. All isoforms of BmTRA-2 protein contain two arginine/serine-rich domains and one RNA recognition motif, showing striking organizational similarity to Drosophila TRA-2 proteins. PMID- 16270152 TI - Positive charges on lysine residues of the extrinsic 18 kDa protein are important to its electrostatic interaction with spinach photosystem II membranes. AB - To determine the contribution of charged amino acids to binding with the photosystem II complex (PSII), the amino or carboxyl groups of the extrinsic 18 kDa protein were modified with N-succinimidyl propionate (NSP) or glycine methyl ester (GME) in the presence of a water-soluble carbodiimide, respectively. Based on isoelectric point shift, 4-10 and 10-14 amino groups were modified in the presence of 2 and 4 mM NSP, respectively. Similarly, 3-4 carboxyl groups were modified by reaction with 100 mM GME. Neutralization of negatively charged carboxyl groups with GME did not alter the binding activity of the extrinsic 18 kDa protein. However, the NSP-modified 18 kDa protein, in which the positively charged amino groups had been modified to uncharged methyl esters, failed to bind with the PSII membrane in the presence of the extrinsic 23 kDa protein. This defect can not be attributed to structural or conformational alterations imposed by chemical modification, as the fluorescence and circular dichroism spectra among native, GME- and NSP-modified extrinsic 18 kDa proteins were similar. Thus, we have concluded that the positive charges of lysyl residues in the extrinsic 18 kDa protein are important for its interaction with PSII membranes in the presence of the extrinsic 23 kDa protein. Furthermore, it was found that the negative charges of carboxyl groups of this protein did not participate in binding with the extrinsic 23 kDa protein associated with PSII membranes. PMID- 16270149 TI - [Renal disease severity index, clinical indicators and mortality of patients in hemodialysis]. AB - OBJECTIVE: Comorbidity is a major factor influencing mortality in hemodialysis patients. Kt/V, hematocrit and albumin levels have also been associated with mortality in these patients. The purpose of this study was to evaluate the severity of comorbidity, Kt/V, hematocrit and albumin levels as predictors of mortality in patients on hemodialysis therapy. METHODS: Forty patients were followed up during 12 months and assessed in relation to social demographic characteristics, time on dialysis therapy, presence of diabetes, Kt/V, hematocrit and albumin levels, also comorbidities. The impact of comorbidity on mortality was assessed by the end-stage renal disease severity index (ESRD-SI). RESULTS: Mean ESRD-SI scores for survivals (85%) and deaths (15%) were 22 +/- 14.8 vs. 44 +/- 12.4 (p < 0.001), and for diabetic (29%) and non-diabetic patients (71%), 40 +/- 15.1 vs. 19 +/- 12.5 (p < 0.001). An inverse correlation was observed between ESRD-SI scores and albumin (r = -0.475; p < 0.005). Albumin levels = 3.6 g/dL were mostly observed (82%) in patients without diabetes (p = 0.021). A correlation was observed between hematocrit and albumin levels (r = 0.544; p < 0.001). For each 1-point increase in the ESRD-SI scores, there was a 10% increase in the risk of death (p = 0.0093). CONCLUSION: The ESRD-SI is useful to assess the severity of comorbidities and to predict mortality in hemodialysis patients. PMID- 16270153 TI - Construction of prophylactic human papillomavirus type 16 L1 capsid protein vaccine delivered by live attenuated Shigella flexneri strain sh42. AB - To express human papillomavirus (HPV) L1 capsid protein in the recombinant strain of Shigella and study the potential of a live attenuated Shigella-based HPV prophylactic vaccine in preventing HPV infection, the icsA/virG fragment of Shigella-based prokaryotic expression plasmid pHS3199 was constructed. HPV type 16 L1 (HPV16L1) gene was inserted into plasmid pHS3199 to form the pHS3199 HPV16L1 construct, and pHS3199-HPV16L1 was electroporated into a live attenuated Shigella strain sh42. Western blotting analysis showed that HPV16L1 could be expressed stably in the recombinant strain sh42-HPV16L1. Sereny test results were negative, which showed that the sh42-HPV16L1 lost virulence. However, the attenuated recombinant strain partially maintained the invasive property as indicated by the HeLa cell infection assay. Specific IgG, IgA antibody against HPV16L1 virus-like particles (VLPs) were detected in the sera, intestinal lavage and vaginal lavage from animals immunized by sh42-HPV16L1. The number of antibody secreting cells in the spleen and draining lymph nodes were increased significantly compared with the control group. Sera from immunized animals inhibited murine hemagglutination induced by HPV16L1 VLPs, which indicated that the candidate vaccine could stimulate an efficient immune response in guinea pig's mucosal sites. This may be an effective strategy for the development of an HPV prophylactic oral vaccine. PMID- 16270154 TI - Identification of immunodominant Th1-type T cell epitopes from Schistosoma japonicum 28 kDa glutathione-S-transferase, a vaccine candidate. AB - Th1-type cytokines produced by the stimulation of Th1-type epitopes derived from defined schistosome-associated antigens are correlated with the development of resistance to the parasite infection. Schistosoma mansoni 28 kDa glutathione-S transferase (Sm28GST), a major detoxification enzyme, has been recognized as a vaccine candidate and a phase II clinical trial has been carried out. Sheep immunized with recombinant Schistosoma japonicum 28GST (Sj28GST) have shown immune protection against the parasite infection. In the present study, six candidate peptides (P1, P2, P3, P4, P7 and P8) from Sj28GST were predicted, using software, to be T cell epitopes, and peptides P5 and P6 were designed by extending five amino acids at the N-terminal and C-terminal of P1, respectively. The peptide 190-211 aa in Sj28GST corresponding to the Th1-type epitope (190-211 aa) identified from Sm28GST was selected and named P9. The nine candidate peptides were synthesized or produced as the fusion protein with thioredoxin in the pET32c(+)/BL21(DE3) system. Their capacity to induce a Th1-type response in vitro was measured using lymphocyte proliferation, cytokine detection experiments and flow cytometry. The results showed that P6 (73-86 aa) generated the strongest stimulation effect on T cells among the nine candidate peptides, and drove the highest level of IFN-gamma and IL-2. Therefore, P6 is a functional Th1-type T cell epitope that is different from that in Sm28GST, and will be useful for the development of effective vaccines which can trigger acquired immunity against S. japonicum. Moreover, our strategy of identifying the Th1-type epitope by a combination of software prediction and experimental confirmation provides a convenient and cost-saving alternative approach to previous methods. PMID- 16270155 TI - Fast fourier transform-based support vector machine for prediction of G-protein coupled receptor subfamilies. AB - Although the sequence information on G-protein coupled receptors (GPCRs) continues to grow, many GPCRs remain orphaned (i.e. ligand specificity unknown) or poorly characterized with little structural information available, so an automated and reliable method is badly needed to facilitate the identification of novel receptors. In this study, a method of fast Fourier transform-based support vector machine has been developed for predicting GPCR subfamilies according to protein's hydrophobicity. In classifying Class B, C, D and F subfamilies, the method achieved an overall Matthe's correlation coefficient and accuracy of 0.95 and 93.3%, respectively, when evaluated using the jackknife test. The method achieved an accuracy of 100% on the Class B independent dataset. The results show that this method can classify GPCR subfamilies as well as their functional classification with high accuracy. A web server implementing the prediction is available at http://chem.scu.edu.cn/blast/Pred-GPCR. PMID- 16270156 TI - Function of c-Fos-like and c-Jun-like proteins on trichostatin A-induced G2/M arrest in Physarum polycephalum. AB - The homologs of transcription factors c-Fos and c-Jun have been detected in slime mold Physarum polycephalum during progression of the synchronous cell cycle. Here we demonstrated that c-Fos-like and c-Jun-like proteins participated in G2/M transition by the regulation of the level of Cyclin B1 protein in P. polycephalum. The study of antibody neutralization revealed that interruption of the functions of c-Fos-like and c-Jun-like proteins resulted in G2/M transition arrest, implicating their functional roles in cell cycle control. When G2/M transition was blocked by histone deacetylase inhibitor trichostatin A, changes in c-Fos- and c-Jun-like protein levels, and hyperacetylation of c-Jun-like protein, were observed. The data suggest that in P. polycephalum, c-Fos- and c Jun-like proteins may be the key factors in the regulation of histone acetylation related G2/M transition, involving the coordinated expression and hyperacetylation of these proteins. PMID- 16270157 TI - Identification of a positive Cis-element upstream of human NKX3.1 gene. AB - NKX3.1 is a prostate-specific homeobox gene related to prostate development and prostate cancer. In this work, we aimed to identify precisely the functional cis element in the 197 bp region (from -1032 to -836 bp) of the NKX3.1 promoter (from -1032 to +8 bp), which was previously identified to present positive regulatory activity on NKX3.1 expression, by deletion mutagenesis analysis and electrophoretic mobility shift assay (EMSA). A 16 bp positive cis-element located between -920 and -905 bp upstream of the NKX3.1 gene was identified by deletion mutation analysis and proved to be a functional positive cis-element by EMSA. It will be important to further study the functions and regulatory mechanisms of this positive cis-element in NKX3.1 gene expression. PMID- 16270158 TI - Knockdown of human p53 gene expression in 293-T cells by retroviral vector mediated short hairpin RNA. AB - RNA interference (RNAi) is an evolutionarily conserved process of gene silencing in multiple organisms, which has become a powerful tool for investigating gene function by reverse genetics. Recently, many groups have reported to use synthesized oligonucleotides or siRNA encoding plasmids to induce RNAi in mammalian cells by transfection, but this is still limited in its application, especially when it is necessary to generate long-term gene silencing in vivo. To circumvent this problem, retrovirus- or lentivirus-delivered RNAi has been developed. Here, we described two retroviral systems for delivering short hairpin RNA (shRNA) transcribed from the H1 promoter. The results showed that retroviral vector-mediated RNAi can substantially downregulate the expression of human p53 in 293-T cells. Furthermore, the retroviral vector-mediated RNAi in our transduction system can stably inactivate the p53 gene for a long time. Compared to shRNAs transcribed from the U6 promoter, H1-driven shRNA also dramatically reduced the expression of p53. The p53 downregulation efficiencies of H1- and U6 driven shRNAs were almost identical. The results indicate that retroviral vector delivered RNAi would be a useful tool in functional genomics and gene therapy. PMID- 16270159 TI - Interaction of mouse Pem protein and cell division cycle 37 homolog. AB - Mouse Pem, a homeobox gene, encodes a protein consisting of 210 amino acid residues. To study the function of mouse Pem protein, we used the yeast two hybrid system to screen the library of 7-day mouse embryo with full-length mouse Pem cDNA. Fifty-two colonies were obtained after 1.57 x 10(8) colonies were screened by nutrition limitation and beta-galactosidase assay. Seven individual insert fragments were obtained from the library, and three of them were identified, one of which was confirmed to be the cell division cycle 37 (Cdc37) homolog gene by sequencing. The interaction between mouse Pem and Cdc37 homolog was then confirmed by glutathione S-transferase pull-down assay, and the possible interaction model was suggested. PMID- 16270160 TI - Stretchin-klp, a novel Drosophila indirect flight muscle protein, has both myosin dependent and independent isoforms. AB - Stretchin-klp is a newly described protein in Drosophila indirect flight muscles (IFM) that migrates on SDS gels as two distinct components of approximately 225 and 231 kD. Although the larger isoform is IFM specific, the smaller stretchin klp isoform is expressed not only in IFM, but also in wild-type tissues of the adult head, abdomen and thorax from which the IFM has been removed. It is not detected, however, in jump or leg muscles. Probes derived from a cDNA encoding part of stretchin-klp hybridize with a 6.7 kb mRNA. Stretchin-klp is one of several putative products of the Stretchin-Myosin light chain kinase gene and is predicted to have multiple immunoglobulin domains arranged in tandem pairs separated by variable length spacers. Polyclonal antibodies directed against the expressed peptide of the stretchin-klp cDNA label the IFM myofibril A-band, though not its central and lateral regions. Analyses of IFM mutants indicate that the larger stretchin-klp isoform is myosin dependent. Although the normal adult myosin filament or the 'headless' myosin rod is sufficient for accumulation of both the large and small stretchin-klp isoforms, loss of myosin, or substitution of the adult rod with an embryonic one in IFM prevents the larger isoform from being formed or stabilized. During development stretchin-klp is first detected at pupal stage p8, when myofibrils are being constructed. These studies suggest that this newly identified protein is a major component of the Drosophila IFM thick filament. PMID- 16270161 TI - The surgical anatomy of the parotid fascia. AB - Currently, there are no reports in the literature of the parotid fascia suggesting that this structure is crucial to the identification of the facial nerve trunk (FNT). Traditional surgical and anatomical descriptions of this fascia report it as a collection of connective tissue large enough to be described by the unaided eye. This study was performed to investigate the composition and limit of the fascia surrounding the parotid gland. An appreciation of these on safe and effective parotid surgery was also considered. HISTO-ANATOMICAL STUDY: Microsurgical step-by-step dissection was performed on 18 adult cadavers (n=36) to define the composition, arrangement, and attachment of the parotid fascia. Samples were subjected to the Masson Trichrome Technique (1990). CLINICAL STUDY: A total of 18 patients presented for parotidectomy. Eight patients had a pleomorphic adenoma and ten had lympho-epithelial disease of the parotid gland. Boundaries of parotid fascia were posteriorly-mastoid process, anteriorly--ramus of mandible, superiorly--cartilage of external acoustic meatus, and inferiorly-imaginary line joining tip of mastoid process to ramus of mandible. These landmarks formed a quadrangular space. HISTO-ANATOMICAL STUDY (N=36): Parotid fascia formed a fibrous meshwork over the gland. In the upper two thirds, fascia was thick and strong; in the lower one-third, fascia was thin. Soft tissue arrangement (from superficial to deep): dermis, subcutaneous fat, superficial cervical fascia, deep cervical fascia. CLINICAL STUDY (N=18): The technique described was applied consistently in all patients. Mean time for localization of FNT was 11 min (range 7-18 min). In two patients (both with an underlying inflammatory disorder of the parotid gland), a transient facial nerve palsy developed postoperatively. In both patients, this settled within 7 weeks of operation. The true surgical potential of the parotid fascia during parotidectomy has been reported. PMID- 16270164 TI - Treatment of rectal cancer without radiotherapy? PMID- 16270162 TI - Thermoresponsive gelatin/monomethoxy poly(ethylene glycol)-poly(D,L-lactide) hydrogels: formulation, characterization, and antibacterial drug delivery. AB - PURPOSE: The primary objective of this study was to prepare novel thermoresponsive binary component hydrogels composed of gelatin and monomethoxy poly(ethylene glycol)-poly(D,L-lactide) (MPEG-PDLLA) diblock copolymer and to obtain optimal formulations capable of forming gels upon a narrow temperature range between body temperature and room temperature. METHODS: MPEG-PDLLA diblock copolymers with a lower critical solution temperature (LCST) feature were synthesized by using a ring-opening polymerization method. The starting weight ratio of MPEG/DLLA was varied to obtain a series of copolymers with a wide range of molecular weight and hydrophilicity. The copolymers were characterized by 1H nuclear magnetic resonance (1H NMR) and thermogravimetric analysis. MPEG (2K) PDLLA (1:4) was chosen to construct hydrogels with gelatin. To obtain optimal thermoresponsive formulation, various hydrogels were formulated and quantified in terms of sol-gel phase transition kinetics and rheological properties. Selected hydrogels were studied as drug carrier for gentamicin sulfate. RESULTS: Gelatin/MPEG-PDLLA hydrogels underwent gelation in less than 15 min when 30 wt.% MPEG (2K)-PDLLA (1:4) was mixed with 10, 50, or 100 mg/mL gelatin. Hydrogels showed rapid gelation when 100 mg/mL gelatin was mixed with 15, 20, or 25 wt.% MPEG-PDLLA as temperature fell from 37 degrees C to room temperature. The viscosity of hydrogels depended on the frequency applied in the rheological tests, the environment temperature, and the concentration of both polymer components. The time needed for 50% gentamicin sulfate release was 5 days or longer at room temperature, and the release lasted up to 40 days. 1H NMR confirmed that MPEG-PDLLA hydrolyzed under in vitro situations. CONCLUSIONS: The incorporation of a second polymer component MPEG-PDLLA into the gelatin hydrogel could modify the thermal characteristic of gelatin and the resulting binary component hydrogels obtained different thermal characteristics from the individual polymer components. Formulation of gelatin/MPEG-PDLLA hydrogels could be varied for obtaining such gels that can undergo gelation promptly upon a narrow temperature change. PMID- 16270165 TI - Ultrastructure of the sensorimotor cortex of pubertal offspring of alcoholic male rats. AB - The ultrastructure of neurons, astrocytes, and capillaries in the sensorimotor cortex of three-month-old offspring of alcoholic male rats (8 g/kg of 50% alcohol solution daily for four weeks) was studied. Apart from signs of delayed maturation of nerve and glial cell processes, some cortical areas showed destructive changes affecting a proportion of neurons along with elements of compensatory-adaptive processes in some nerve cells. Membrane and myelin-like intranuclear inclusions and changes in the Golgi complex were characteristic features of the damage to populations of cortical neurons. Swelling of astrocytes, deformation of capillaries and changes in their ultrastructure, with accumulation of atypical inclusions in pericapillary astrocyte processes, degraded reciprocal exchange processes between blood and neurons. The possible role of the ischemic-hypoxic factor in delayed changes in the ultrastructure of the sensorimotor cortex in the offspring of alcoholic males is discussed. PMID- 16270166 TI - Organization of efferent projections of the pedunculopontine nucleus of the tegmentum of the striatum in the dog brain. AB - A method based on retrograde axonal transport was used to undertake detailed studies of the efferent projections of individual substructures of the pedunculopontine nucleus of the midbrain tegmentum to functionally diverse segments of striatal structures in dogs. The compact and diffuse zones of this nucleus were found to project to segments of the putamen, caudate nucleus, and nucleus accumbens, which are components of the limbic and motor systems. The medial part of the diffuse zone, adjacent to the decussation of the superior cerebellar peduncles (presumptively the mesencephalic extrapyramidal field) projected only to the dorsal segments of the caudate nucleus and putamen, which are parts of the motor system. PMID- 16270167 TI - Gaba- and serotonin-immunoreactive structures and ca(2+)-binding protein in the neocortex of the reeler mouse mutant. AB - The transmitter organization of anomalously formed neocortex was studied in reeler mutant mice by immunohistochemical studies of GABA- and serotoninergic structures and Ca(2+)-binding protein. GABAergic structures were identified in terms of the localization of glutamate decarboxylase (GDC) within them, this being an enzyme involved in GABA synthesis. The neocortex of reeler mutant mice was found to contain an unusual distribution of cells morphologically and chemically identical to Cajal-Retzius cells - beneath layer I rather than in its upper third, as seen in normal animals. GDC-immunoreactive label accumulated in the neuropil of the intermediate and deep layers, layer I containing only occasional granules. Serotonin-immunoreactive fibers did not form superficial or deep plexuses, as seen in normal animals, though they did reach their innervation targets. Thus, the anomalously formed neocortex which lacks the typical cytoarchitectonic organization, showed abnormalities in the structure of both intrinsic and projectional transmitter systems. PMID- 16270169 TI - Morphocytochemical analysis of neuron populations in the ganglion layer of the cerebellar cortex of birds and mammals living in similar habitats. AB - The neuron populations in the ganglion layer of the cerebellar cortex of birds and mammals living in similar habitats, i.e., the common squirrel and the house sparrow (Sciurus vulgaris and Passer domesticus), living in an air/land habitat, and the swamp beaver (nutria) and domestic goose (Myocastor coypus and Anser domesticus), living in an earth-water habitat, were studied. Morphocytochemical measures were assessed, i.e., the density of Purkinje cells per unit gyrus length and their sizes, staining properties, and protein contents, for neuron populations in the ganglion layer. Assessment of the staining properties of neurons revealed general features in the organization of the ganglion layer of the cerebellum of vertebrates; assessment of protein contents demonstrated specific features typical for particular classes independently of the animals' habitats. PMID- 16270168 TI - Nestin-expressing cells in the human hippocampus. AB - Nestin, a protein of the intermediate filament family, is typical of undifferentiated neural stem and progenitor cells. The present report describes studies of nestin expression in the hippocampus of patients with epilepsy and identifies five types of nestin-immunopositive cells differing in terms of their morphological phenotype and immunological characteristics. These were cells with the phenotype of radial glial cells, bipolar cells, small dendritic cells, cells of the subependymal zone, and astrocyte-like cells. Two types of cell - radial gliocytes of the dentate fascia and NG2-immunopositive bipolar cells - can be regarded as neural precursors of different levels of commitment. PMID- 16270171 TI - Characteristics of mast cells in the choroid plexus of the ventricles of the human brain in aging. PMID- 16270170 TI - Morphological characteristics of the retinomotor response in salmon trout (oncorhynchus masou) fry in a magnetic field and red light. AB - The retinomotor response was studied in fry of the salmon trout Oncorhynchus masou during experimental exposures to a constant magnetic field and red light alone and together. The responses of photoreceptors and the pigmented epithelium to red light were mesopic in nature. The mesopic state of the retina after exposure of fish to a magnetic field in the dark differed from the pigment epithelium response after exposure to red light. On exposure to the magnetic field after red light, the effects of these two treatments were additive. Rods adapted to low-light conditions, while cones adapted to light. The simultaneous operation of these two mechanisms of perception is thus possible, although it never occurs in normal conditions. On exposure to red light after the magnetic field, the retinomotor response reflected a physiological dysfunction in which neither rods nor cones were operative. The pigmented epithelium is actively involved in responses to changes in the magnetic field. It is suggested that cells of the pigmented epithelium can function as light-sensitive magnetoreceptors. PMID- 16270172 TI - Studies of reconciliation in anubis baboons. AB - Studies of post-conflict reconciliation were performed on anubis baboons living in corrals. Reconciliation was found to occur after more than a third of the conflicts observed; the mean reconciliatory tendency was 27.3 +/- 2.4. Different categories of conflict pairs showed similar reconciliation frequencies. In 75% of cases, reconciliation occurred within the first 2 min of completion of the conflict. The set of behavioral fragments demonstrated by participants in reconciliation depended on gender and role in the conflict. Differences were seen in the magnitudes of the reconciliatory tendencies of monkeys of high and low rankings. PMID- 16270173 TI - A model of anxious depression: persistence of behavioral pathology. AB - Chronic psychoemotional stress induced by negative experience of social defeats in intermale confrontations over a period of 30 days was found to lead to the development of anxious-depressive symptomatology in male mice. Cessation of the psychopathogenic conditions and placing of depressed animals in comfortable conditions for 1-2 weeks with females did not lift the pathological state. Individuals continued to show marked anxiety, a behavioral deficit, decreased communicativeness, and a high level of depressivity, as revealed by a variety of behavioral tests. Persistence of the resulting psychoemotional disturbance in these animals is evidence for the development and persistence of the behavioral pathology requiring drug treatment. PMID- 16270176 TI - Changes in the excitability of pyramidal neurons in hippocampal slices from aged rats in conditions of periodic transient episodes of hypoxia. PMID- 16270175 TI - Effects of prenatal stress on the activity of an enzyme involved in neurosteroid synthesis during the "critical period" of sexual differentiation of the brain in male rats. AB - The effects of daily immobilization stress applied to female rats on days 15 to 18 of pregnancy on the activity of the enzyme 5 alpha-reductase (isoform I), involved in the synthesis of brain neurosteroids were studied in male offspring. The results demonstrated a decrease in enzyme activity in the cerebral cortex and hypothalamus of male fetuses one day after the last session of stress, while enzyme activity was elevated in the cortex of neonates. Increases in 5 alpha reductase activity in the cortex, hippocampus, and hypothalamus were also seen in prenatally stressed males on day 5 of life. There were reductions in plasma testosterone and progesterone levels in experimental animals on day 19 of embryonic life and in neonatal rats, the blood progesterone level in prenatally stressed rats remaining decreased at age five days. The possible involvement of neurosteroids in the actions of prenatal stress on sexual differentiation of the brain is discussed. PMID- 16270174 TI - Sequelae of prenatal serotonin depletion and stress on pain sensitivity in rats. AB - The effects of interrupted synthesis of serotonin (5-HT) and immobilization stress applied to pregnant Wistar rats on behavioral measures of pain sensitivity in the formalin test were studied in their offspring at age 90 days. Prenatal 5 HT depletion decreased pain sensitivity in one third of rats and produced no significant change in the remainder. However, the latter showed a clear tendency to an increase in the interphase interval in females and a decrease in males. Prenatal stress increased pain sensitivity in 50% of rats with prenatal 5-HT deficiency and decreased pain sensitivity in the remainder. Increases in pain sensitivity were also seen in control rats (with prenatal injections of physiological saline), though to a significantly lesser extent than in animals with prenatal 5-HT depletion. The latter showed gender differences in the effects of prenatal stress on pain sensitivity. The present data provide evidence that prenatal 5-HT depletion has long-term effects on the functional activity of the nociceptive system and the important role of 5-HT in mediating the effects of prenatal stress on pain sensitivity in the formalin test. PMID- 16270177 TI - Neuron activity in the monkey striatum of identifies integration sequential actions into functional blocks. AB - Spike activity in monkey striatum (putamen) neurons was recorded during the performance of a complex multistep operant task. Tonic responses propagating beyond a single action were recorded, along with phasic responses seen within a given action. The tonic type of response was recorded in 132 of 148 cells. Only 11 of these neurons showed exclusively this type of activity. The beginnings and ends of tonic responses were generally associated with key moments in the behavior, corresponding to the triggering and completion of immediate aims during the performance of the behavioral program as a whole. These results provide evidence that the role of the striatum is not limited to controlling single sequentially performed actions, but spreads to the whole structure of a behavioral act. PMID- 16270179 TI - Formation of behavioral pathology in female C57BL/6J mice exposed to prolonged negative psychoemotional conditions. AB - Prolonged exposure of females to negative psychoemotional conditions by permanent keeping with aggressive males on the other side of a perforated partition in a cage with daily witnessing of intermale confrontations had negative influences on the behavior of females, assessed using a variety of ethological tests. The elevated maze test and the open field test provided evidence for the development of marked anxiety in the females, with inhibition of motor and investigative activity. Decreases in the responses of females close to the partition to familiar and unfamiliar males one month after the onset of exposure to negative psychoemotional conditions and before the end of the experiment provided evidence for a decrease in communicativeness. After two months of exposure to negative psychoemotional conditions, intermale confrontations also ceased to interest females, as shown by the partition test. The hotplate test revealed worsening of pain sensitivity in experimental females. More than half of the individuals demonstrated impairment to the cyclicity of sexual cycles with increases in the duration of diestrus. After being kept in negative psychoemotional conditions, females ceased to differentiate littermates by social status and gender, as shown by social/sexual recognition tests. All these data provide grounds for suggesting the development of an anxious-depressive state in females after being kept in negative psychoemotional conditions for prolonged periods. PMID- 16270178 TI - Analysis of evoked EEG synchronization and desynchronization during perception of emotiogenic stimuli: association with autonomic activation processes. AB - The cortical apparatus involved in performing autonomic responses in conditions of emotional activation has received little study. The aim of the present work was to assess the dynamics of evoked EEG synchronization and desynchronization at different frequency ranges during the perception of emotiogenic visual stimuli depending on the extent of accompanying autonomic activation as measured by skin galvanic responses. Studies were performed on 33 students (all right-handed) aged 18-28 years. Difference between subjects with weak (SGR(-)) and strong (SGR(+)) skin galvanic responses were seen only in the theta1 range (4-6 Hz). At the stage at which emotiogenic information was perceived (the first second after the start of stimulus presentation), both groups showed similar dynamics of increases in evoked synchronization in the parietal-temporal-occipital areas of the cortex, with greater involvement of the right hemisphere. From the second second to the end of presentation (2-6 sec), emotiogenic signals gave significantly greater levels of evoked synchronization in these cortical areas as compared with neutral stimuli, and only in the SGR(+) group. These data provide evidence for the involvement of the posterior areas of the cortex of the right hemisphere in the mechanisms of motivational attention and sympathetic activation. PMID- 16270180 TI - Transcranial micropolarization in the combined therapy of speech and general psychomotor retardation in children of late preschool age. AB - The efficacy of a method of combined treatment, developed by ourselves, using transcranial micropolarization in patients with delayed development of speech and general mental development, was studied in 30 children aged 4-6 years. High therapeutic effects were seen with this method in 60-80% of children, with accelerated development of communicative behavior and formation of speech-motor functions, along with increases in the stability of attention and improvements in the processes of perception and the productivity of cognitive activity. PMID- 16270181 TI - The current state of S. S. Korsakov's concept of alcoholic polyneuritic psychosis. PMID- 16270183 TI - Anharmonic compression of the glitter lattice. AB - A previous report has described the crystal structure of glitter, which is a dense 3-,4-connected net composed of ethylenic columns that run parallel to the c axis of the unit cell. Such a structure invites speculation as to its relative stiffness along that axis. A semiempirical expression due to Cohen was used in a previous communication to estimate its zero-pressure bulk modulus. This estimate exceeds that of any known material at 440 GPa. Further, by treating the ethylenic units as harmonic springs, a correction was computed for the elastic deformation of the carbon-carbon double bonds along the c-axis. This correction is on the order of 300 GPa for deformations of the double bonds of approximately 0.1 angstroms. The present communication treats the ethylenic units along the c-axis of glitter as anharmonic springs obeying a Morse potential and a Morse's law force. Within the anharmonic approximation, at modest bond length deformations, x', the bulk modulus at pressure of the glitter lattice exceeds 1 TPa. PMID- 16270182 TI - Non-stochastic and stochastic linear indices of the molecular pseudograph's atom adjacency matrix: a novel approach for computational in silico screening and "rational" selection of new lead antibacterial agents. AB - A novel approach (TOMOCOMD-CARDD) to computer-aided "rational" drug design is illustrated. This approach is based on the calculation of the non-stochastic and stochastic linear indices of the molecular pseudograph's atom-adjacency matrix representing molecular structures. These TOMOCOMD-CARDD descriptors are introduced for the computational (virtual) screening and "rational" selection of new lead antibacterial agents using linear discrimination analysis. The two structure-based antibacterial-activity classification models, including non stochastic and stochastic indices, classify correctly 91.61% and 90.75%, respectively, of 1525 chemicals in training sets. These models show high Matthews correlation coefficients (MCC=0.84 and 0.82). An external validation process was carried out to assess the robustness and predictive power of the model obtained. These QSAR models permit the correct classification of 91.49% and 89.31% of 505 compounds in an external test set, yielding MCCs of 0.84 and 0.79, respectively. The TOMOCOMD-CARDD approach compares satisfactorily with respect to nine of the most useful models for antimicrobial selection reported to date. Finally, an in silico screening of 87 new chemicals reported in the anti-infective field with antibacterial activities is developed showing the ability of the TOMOCOMD-CARDD models to identify new lead antibacterial compounds. PMID- 16270184 TI - [Use of fragrances. What about the side effects?]. AB - Fragrances are increasingly used in private and public domains. Over recent years the olfactory sense has been paid more and more scientific and economic attention. While on the one hand bad smells are counteracted by fragrances, marketing experts are now trying to introduce this sense into multimedia-based experiences. Technical means are used to address positively and directly the sense of smell. The aim is to make the smell a unique feature for a certain brand or location. When it comes to "style of living" or "special shopping experience" nowadays the olfactory design plays an important role. Although fragrances are applied very frequently, there is still a lack of knowledge about the potential consequences for health and the environment. Certain substances (musk compounds) have been proven persistent and accumulative, and others belong to the most common causes of contact eczema. Some people also report special sensitivities towards certain smells for unknown reasons. Unlike audiovisual attractions it is very difficult for humans to avoid olfactory stimuli. The question arises whether fragrance materials constitute a group of substances that should receive more attention concerning their risk for health and the environment. PMID- 16270185 TI - [Sport and health among adults in Germany]. AB - Regularly participating in sports activities can have a positive effect on people's health and is therefore an important area of health promotion. Data of the Telephone Health Survey 2003 indicate that until now there is an unexploited potential for programmes promoting sports: in total 37.3% of men aged 18 years and above and 38.4% of equally aged women do not participate in sports at all; another 20.9% of men and 28.4% of women only engage in sports activities for up to 2 h/week. The highest prevalence of sports activity is seen among young adults, whereas it is lowest among older people. However, the latter slightly increased their sports activity during the last 10 years, while the trend observed among younger adults seems to point rather in the opposite direction. Also relevant for the planning and implementation of group-specific interventions is the observation that persons with low socioeconomic status tend to engage less often in sports than people with middle or high status do. The present study underlines the health relevance of sports participation by presenting inverse relationships between sports activity and behaviour-related health risks such as smoking and obesity as well as a positive effect of sports on self-assessed health. PMID- 16270186 TI - [Measuring health-related quality of life with the SF-8. Normal sample of the German population]. AB - The SF-8, a relatively new instrument for measuring health-related quality of life, was used in the German National Telephone Health Survey 2003. The SF-8 provides results which are comparable to those of the SF-36, the instrument most commonly used internationally. The German National Telephone Health Survey provides nationally representative data for the residential population in Germany aged 18 and older. In addition to the measurement of health-related quality of life, comprehensive information on chronic illnesses and complaints, health care needs, utilisation of health care, risk factors, risk behaviour and social status is also collected, making differentiated analyses possible. According to the data collected, men rate their quality of life in all dimensions higher than women. With increasing age, quality of life for both men and women decreases in the physical dimensions, while increasing in the mental health dimensions. Apart from chronic disease and pain having a negative impact on health-related quality of life, social differences are also observable, in that worse health-related quality of life is reported by respondents with lower socioeconomic status. PMID- 16270187 TI - [Public health care utilisation. Initial results of the Telephone Health Survey 2003]. AB - More than 90% of adults have a family doctor. The more chronic diseases and the more subjective health problems the interviewees had, the more often they had been to see their family doctor. Senior citizens and people with lower social status or a higher BMI go to see their family doctors more frequently. People who say they consume a lot of alcohol consult a doctor less often than others. Privately insured people also go to see their family doctors less frequently. Almost two-thirds of the adult population in Germany have seen a doctor in the last 3 months, and 44% of people over 60 had an influenza vaccination this year. People are much more willing to have an influenza injection in eastern Germany than in the west. More than half of the people questioned over the age of 35 has had a general health check-up at least once in their lives. Men have health check ups slightly more often than women do. Of the women, 80% have participated at least once in a cancer screening programme, and as many as 60% of women had a cancer scan during the last year. Men take up cancer screening offers less often than women do. Inpatients stay for an average of 2 weeks in hospital per year. Just under 30% of those questioned have had medical rehabilitation treatment. PMID- 16270188 TI - [Overweight and obesity in Germany 1984-2003]. AB - The magnitude and the development of the prevalence of overweight and obesity in Germany during the past 20 years are analysed in this contribution. Using body mass index (BMI) data of the National Examination Surveys (1984-1992), the German National Health Interview and Examination Survey 1998 as well as the Telephone Health Survey 2003, all representative of the adult non-institutionalised population, the occurrence of overweight and obesity is studied. To improve the comparability with measurement values, the Telephone Health Survey data which were based on self-reporting were adjusted. All data were weighted to improve the representativeness for the time of assessment. Although the proportion of persons with a BMI between 25 and 30 did not change essentially during the last 20 years, the proportion of obese individuals (BMI>or=30) increased considerably. At present, about 70% of men and 50% of women in Germany are overweight or obese. In conclusion, the prevalence of overweight and obesity in Germany continues to increase. PMID- 16270189 TI - [Postoperative CT-controlled results of renewed osteosynthesis using screw fixation for acute scaphoid fractures]. AB - BACKGROUND: Currently screw fixation of the scaphoid is a well-established method to treat unstable scaphoid fractures. PATIENTS AND METHODS: Between June 1995 and December 2000, 68 patients with an unstable acute scaphoid fracture were treated with screw fixation; 46 patients were reexamined on an average 35 months postoperatively. Range of motion and grip strength (Jamar dynamometer) were measured. The total data rating resulted from the Krimmer wrist score. RESULTS: The subjective results were evaluated with the DASH questionnaire. The average postoperative pain score was documented with a visual analogous pain scale from zero to 100 (VAS 0-100). X-rays as well as computed tomography were performed postoperatively. The average range of motion was 124 degrees for extension/flexion (92% of the opposite site), 57 degrees for radial/ulnarduction (=90%), and 177 degrees for pronation/supination (=98%). Postoperative strength was 47 kg (=90% of the opposite site). The postoperative pain score was 13 (0 100) after stress and 2 during resting conditions. Bony consolidation was reached in 44 cases. The Krimmer wrist score demonstrated a very good result in 39 cases, a good result in 5 cases, and a satisfactory result in 3 cases. The average DASH score was 8.3 points. CONCLUSIONS: The results demonstrate the reliability of internal screw fixation as treatment for acute scaphoid fractures. Functional results as well as subjective satisfaction of the patients are very good. Postoperative CT scans help to evaluate exact bony consolidation, position of the screw, and postoperative morphology of the scaphoid. PMID- 16270190 TI - [Tension pneumopericardium--a rare complication in multiply injured patients]. AB - Thoracic injury is a relevant and common complication in multiply injured patients. Typical patterns of injury comprise rib fractures, serious lung trauma as well as diaphragmatic and aortic rupture. In contrast, posttraumatic tension pneumopericardium following blunt thoracic trauma is a very rare complication. However, if unrecognized it might provoke cardiac tamponade and death. For the development of a pneumopericardium, free air follows the vessel bundles up to the pericardium. Hence, if the number of ruptured alveoli is high, or these alveoli are placed close to the heart, and if additional risk factors, such as high inspiratory ventilation pressure, are present, a tension pneumopericardium can induce cardiac tamponade. The aim of this report is to illuminate diagnostic and therapeutic strategies for posttraumatic pneumopericardium by presentation of a case from our trauma centre and a critical discussion of the present literature. PMID- 16270191 TI - Medium-term effects of chemotherapy in older cancer patients. AB - PURPOSE: To address the lack of research in older cancer patients, the present study prospectively evaluated their cognitive functions across the first six months following diagnosis and chemotherapy. PATIENTS AND METHODS: A total of 77 younger (n=43, age < 60) and older (n=34, age > or = 60) cancer patients with hematological disease or cancer of the intestinal tract took part in the study. Medium-term effects of chemotherapy were examined in these cancer patients by means of a battery of cognitive tests during baseline and six months after start of treatment. RESULTS: In contrast to baseline verbal learning, word fluency and memory capacity improved in all patients six months after start of treatment. A negative effect of age on cognitive functions could not be demonstrated. Additionally, depression was neither associated with medium-term effects of chemotherapy nor was it related to age. CONCLUSION: The results suggest that chemotherapy has no negative effects on cognitive functions after the first six months following the onset of treatment. It was demonstrated that complete recovery of cognitive functions is independent of the patient's age. PMID- 16270192 TI - EuroQol and survival prediction in terminal cancer patients: a multicenter prospective study in hospice-palliative care units. AB - GOALS OF WORK: Although the EuroQol (EQ-5D) is widely used for economic evaluation, it remains unclear whether it can be combined with medical data to predict survival in patients with terminal cancer. PATIENTS AND METHODS: We carried out this prospective study on 142 terminal cancer patients in four hospice-palliative care units. Association was sought between survival time and a range of variables such as cancer site, performance, previous treatment, age, sex, pain, and EuroQol. The EQ-5D was transformed into the corresponding EQ-5D utility. For univariate analysis, we estimated differences in survival with the Gehan generalized Wilcoxon test. For those variables that were significant, we performed multivariate analysis using the Cox proportional hazard model. MAIN RESULTS: Univariate analysis showed that sex, age, performance, previous use of chemotherapy, and the EQ-5D utility provided statistically significant prognostic survival information. The median survival time was 13.0 days for the group with an EQ-5D utility score lower than -0.5 and 21.0 days for the group with an EQ-5D utility score above -0.5. In multivariate analysis with the Cox proportional hazard model, an EQ-5D utility score < or = 0.5 (RR 1.57, 95% confidence interval 1.06-2.33) was an independent negative predictor of survival. CONCLUSIONS: The EQ 5D quality-of-life assessment tool might be useful for predicting survival time for terminal cancer patients. PMID- 16270193 TI - Infections in a pediatric patient cohort with acute lymphoblastic leukemia during the entire course of treatment. AB - GOALS: To assess the type, frequency, severity, and outcome of all infectious episodes in a pediatric patient cohort with acute lymphoblastic leukemia (ALL) from a single institution during the entire length of leukemia treatment. PATIENTS AND METHODS: Eighty-six patients were treated according to a modified ALL Berlin-Frankfurt-Munster protocol. Retrospective analysis of all types of infections according to the treatment phase and the degree of neutropenia is presented. RESULTS: A total of 610 infectious episodes were recorded. Most infections were documented during maintenance (57%), followed by the induction phase (20.3%). During maintenance, 347 episodes were encountered, with nonspecific viral upper respiratory tract infections (URIs) being the commonest diagnosis (40.0%). Additionally, 38 of 58 total specific viral infections occurred during maintenance: 16 herpes simplex, 7 varicella, 10 herpes zoster infections [varicella-zoster virus (VZV), 45%]. The majority of bacteremia and fever of unknown origin occurred during induction (20%). The number of Gram negative bacteremia was 50% of the total (26 of 52). The majority of the infections (59.5%) occurred without neutropenia [absolute neutrophil count (ANC) >1,000 microl(-1)]. Fewer infections (9.3%) were recorded with concurrent very severe neutropenia (ANC <100 microl(-1)), although 38.5% of positive blood cultures were documented with severe neutropenia. No infection-related fatality occurred. CONCLUSIONS: Most of the severe infections occurred during induction. Gram-positive bacteremia and Gram-negative bacteremia were almost equal. URIs were the commonest infections during the entire treatment and during maintenance. Specific viral infections represented a smaller percentage of the total (VZV was the commonest pathogen). Infectious complications represented a significant morbidity factor, but notably, mortality was negligible. PMID- 16270195 TI - Fetal and neonatal exposure to nicotine in Wistar rats results in increased beta cell apoptosis at birth and postnatal endocrine and metabolic changes associated with type 2 diabetes. AB - AIMS/HYPOTHESIS: Epidemiological studies report an increased risk of obesity and type 2 diabetes in children born to women who smoked during pregnancy. This study examines the effect of fetal and neonatal exposure to nicotine, the major addictive component of cigarettes, on postnatal growth, adiposity and glucose homeostasis. METHODS: Female Wistar rats were given either saline (vehicle) or nicotine (1 mg kg(-1) day(-1)) during pregnancy and lactation. Serum and pancreas tissue were collected from the infant rats at birth. Postnatal growth was assessed weekly until the rats reached 26 weeks of age and glucose homeostasis was examined by OGTTs performed at 7 and 26 weeks of age. RESULTS: Exposure to nicotine resulted in increased postnatal growth and adiposity. Nicotine exposure also resulted in dysglycaemia at 7 and 26 weeks of age. Serum insulin concentrations were decreased in the pups exposed to nicotine at birth. This was associated with increased beta cell apoptosis (pups of saline-treated mothers 8.8+/-1.21% apoptotic beta cells; pups of nicotine-treated mothers 27.8+/-3.1% apoptotic beta cells). CONCLUSIONS/INTERPRETATION: Fetal and neonatal exposure to nicotine results in metabolic changes in the offspring that are consistent with obesity and type 2 diabetes. We propose that these metabolic changes may be a consequence of the initial insult to the beta cell during fetal life and that this animal model has many characteristics of diabetes in humans. PMID- 16270197 TI - A fluorescence-based microplate assay to quantify DOM-induced catabolic activity. AB - This note describes a novel method to quickly quantify the dissolved organic matter (DOM)-induced catabolic activity from low-volume samples. The concept is based on the catabolic response profiles (CRP) assay and is described as an inverse CRP, where the reactivity of a complex and diverse mixture of organic compounds towards single strains of bacteria is quantified. A strain of Pseudomonas fluorescens was grown and then transferred to an organic carbon-free mineral salt medium. 90 microL of a fluorogenic redox indicator was added to 90 microL of the bacterial suspension in a well on a 96-well microplate. The DOM sample (90 microL) was added to the well and the fluorescence emitted by the reduced indicator was read over the period of incubation. Only 0.8 mL of the DOM sample, including controls and replicates, was required to quantify the activity of each sample. Results are presented for a surface soil DOM sample and they were compared to glucose samples of various concentrations. The detection limit was reached for samples containing as little as 55 microM of glucose (0.3 mg C L( 1)). The assay showed that only 9% of the total carbon of the soil surface DOM sample was readily biodegradable. PMID- 16270196 TI - Comment to: Grassi G, Dell'oro R, Quarti-Trevano F et al (2005) neuroadrenergic and reflex abnormalities in patients with metabolic syndrome. Diabetologia 48:1359-1365. PMID- 16270194 TI - Glomerular expression of thrombospondin-1, transforming growth factor beta and connective tissue growth factor at different stages of diabetic nephropathy and their interdependent roles in mesangial response to diabetic stimuli. AB - AIMS/HYPOTHESIS: We quantified the glomerular expression of thrombospondin-1 (THBS1, also known as TSP-1), transforming growth factor beta 1 (TGFB1, also known as TGF-beta1) and connective tissue growth factor (CTGF) at each stage of diabetic nephropathy. We also examined the roles of THBS1 and CTGF in mediating high-glucose- and glycated-albumin-induced synthesis of the matrix protein, fibronectin, by mesangial cells. METHODS: THBS1, latent and active TGFB1, and CTGF, were detected by immunohistochemistry and in situ hybridisation in biopsies from 19 insulin-dependent diabetic patients with incipient, manifest and advanced diabetic nephropathy, and in 11 control kidneys. Findings were quantified by image analysis. Human mesangial cells were cultured with normal or high glucose, albumin or glycated albumin (Amadori product), +/-THBS1 or CTGF antisense oligonucleotides, or with peptide W, an inhibitor of TGFB1 bioactivation by THBS1. Proteins were measured by western blot analysis or ELISA. RESULTS: In glomeruli of normal kidneys, mRNA and protein levels for THBS1, latent-TGFB1 and CTGF were low. They were increased in the incipient stage of diabetic nephropathy, predominantly in mesangial areas, with further increases at later stages of the disease. Little or no active TGFB1 immunostaining was detected prior to manifest diabetic nephropathy. In contrast to high-glucose conditions, increases in fibronectin synthesis that were stimulated by glycated albumin were not dependent on THBS1 activation of latent TGFB1. However, increased fibronectin synthesis in both conditions required CTGF. CONCLUSIONS/INTERPRETATION: Increased glomerular expression of all three factors occurs from the earliest stage of diabetic nephropathy. In contrast to THBS1, CTGF is required for mesangial synthesis of fibronectin stimulated by high glucose or glycated albumin, and is thus a potential therapeutic target. PMID- 16270198 TI - [The reorientational rearthrodesis of the upper ankle joint following failed arthrodesis]. AB - There are three important principles for the correction of nonunion and/or malunion of the ankle joint: (1) reorientation back to anatomic shape and to the normal biomechanical axis of the ankle and foot; (2) respect for the biology of bone by resecting all sclerotic bone and/or transplantation of autogenous bone graft; and (3) achievement of optimal biomechanical stability by using the four screw technique, a limited-contact dynamic-compression plate or a blade plate. CT scanning is the most reliable method for detecting nonunion of the ankle joint after arthrodesis. According to Saltzman, in order to understand the pathology of malunions and nonunions and to plan their correction, weight-bearing anteroposterior radiographs with a 20 degrees internal rotation of the feet, precise lateral views, and rear views of both sides are highly recommended. PMID- 16270200 TI - Computational analysis of biomechanical contributors to possible endovascular graft failure. AB - This paper evaluates numerically coupled blood flow and wall structure interactions in a representative stented abdominal aortic aneurysm (AAA) model, leading potentially to endovascular graft (EVG) failure. A total of 12 biomechanical contributors to possible EVG migration were considered. The results show that after EVG insertion for the given model, the peak AAA sac-pressure was reduced to 14.2 mmHg (11.8% of p(lumen)), and hence the maximum von Mises wall stress and wall deformation dropped by factors of 20 and 10, respectively. Thus, an EVG can significantly reduce sac pressure, mechanical stress, pulsatile wall motion, and the maximum diameter in AAAs and hence prevent AAA rupture effectively. In the absence of endoleaks, elevated sac-pressure can still be caused by fluid-structure interactions between the EVG, stagnant blood, and AAA wall. EVG migration forces vary from 1.4 to 7 N for different EVG geometries, material properties, and hemodynamic conditions. AAA-neck angle, iliac bifurcation angle, neck aorta-to-iliac diameter ratio, EVG size, aorto-uni-iliac EVG, and hypertension play important roles in generating forces potentially leading to EVG migration. PMID- 16270199 TI - Bird breeder's disease: a rare diagnosis in young children. AB - Bird breeder's lung disease is the most common form of hypersensitivity pneumonitis and is a rare entity in young children. We report three cases of children under 7 years of age in whom this diagnosis was confirmed early in the course of the disease. Three children aged 4.4 to 6.5 years presented with dry cough lasting for more than 1 month, dyspnoea, variable loss of appetite, weight loss, fatigue, fever and mild signs of respiratory distress. Chest X-ray films and CT scans showed a bilateral micronodular infiltrate. All three patients had strongly suggestive bronchoalveolar lavage fluid findings with lymphocytosis; two had elevated cell counts and decreased CD4/CD8 ratios. Lung biopsy confirmed the diagnosis in all children. Contact with allergens was identified in all children: two had spent holidays close to a farm in the previous month and one was living next to a pigeon house. In all children, avian precipitins were positive. The symptoms rapidly resolved after allergen avoidance and treatment with oral prednisone. Corticoid treatment was given between 11 and 15 weeks. One child relapsed and required long-term low-dose corticotherapy for 1 year. Lung function tests were normal in all three patients, 3.9 to 5.7 years after diagnosis. CONCLUSION: Bird breeder's lung disease is a rare entity but should be considered in young children presenting long lasting cough. While rapid allergen exclusion and start of treatment can avoid the evolution into irreversible lung fibrosis, clinical and biological evolution should be monitored carefully even after stopping corticoid treatment because of the possibility of relapse. PMID- 16270202 TI - Validation, reliability, and complications of a tethering scoliosis model in the rabbit. AB - This study was conducted to refine a small animal model of scoliosis, and to quantify the deformities throughout its growth period. Subcutaneous scapula-to contralateral pelvis tethering surgery was selected due to its minimally invasive nature and potential applicability for a large animal model. The procedure was performed in 7-week-old New Zealand white rabbits. Group A animals (n=9) underwent the tethering procedure with a suture that spontaneously released. Group B animals (n=17) had the identical procedure with a robust tether and pelvic fixation, which was maintained for 2 months during growth. All animals developed immediate post-operative scoliosis with a Cobb angle of 23 degrees (range, 6-39 degrees) in group A and 59 degrees (range, 24-90 degrees) in group B animals. During the 2 month post-tethering, group A animals lost their tether and scoliosis resolved, whereas all animals in group B maintained their tether until scheduled release at which time the mean scoliosis was 62 degrees. Immediately after tether release, group B scoliosis decreased to a mean 53 degrees. Over the following 4 months of adolescent growth, the scoliosis decreased to a mean of 43 degrees at skeletal maturity; the decrease usually occurred in animals with less than 45 degrees curves at tether release. Radiographs revealed apical vertebral wedging (mean 19 degrees ) in all group B animals. Sagittal spinal alignment was also assessed, and for group B animals, the scoliotic segment developed mild to moderate kyphosis (mean 28 degrees) and torsional deformity, but the kyphosis resolved by 4 months after tether-release. Complications specific to this technique included a high rate of transient scapulothoracic dissociation and cases of cor pulmonale. In conclusion, this tethering technique in immature rabbits consistently produced scoliosis with vertebral wedging when the tether was intact through the first 2 months of the protocol. The transient exaggeration of kyphosis suggests that the production of scoliosis is not necessarily dependent on lordosis in this model. Because this technique does not violate thoracic or spinal tissues, it may be useful in the investigation of secondary physiologic effects of mechanically-induced scoliosis, and may be scalable to larger animal species. PMID- 16270203 TI - [Calcinosis cutis in Albright hereditary osteodystrophy: pseudohypoparathyroidism type Ia]. AB - Albright hereditary osteodystrophy (AHO) is characterized by a symptom complex including short stature, brachymetacarpia, obesity, round facies, cutaneous osteomas, and mental retardation. AHO is caused by mutations in the GNAS-gene localized on chromosome 20 encoding for Gsalpha protein, a signal transducer of endocrine pathways. Therefore, AHO is often associated with endocrinopathy such as pseudohypoparathyroidism or hypothyroidism. A nine-month-old boy presented with typical features of this syndrome. The diagnosis was confirmed by biochemical and molecular analyses. An unusual feature was calcinosis cutis at such an early age, which led to extensive differential diagnostic procedures. PMID- 16270201 TI - Calcium transport in human salivary glands: a proposed model of calcium secretion into saliva. AB - Salivary calcium plays a vital role in bio-mineralization of dental enamel and exposed dentin. In order to elucidate the yet unknown cellular and molecular mechanisms of calcium secretion in human salivary glands the presence of various relevant plasma membrane transport systems for calcium were investigated. Using an RT-PCR approach, expression of the epithelial calcium channel (CaT-Like), the calcium binding protein (calbindin-2), the endoplasmic reticulum pumps (SERCA-2 and -3), and the plasma membrane calcium ATPases (PMCA-1, -2, and -4), were found in parotid and submandibular glands. Immunohistochemistry revealed that CaT-Like is located in the basolateral plasma membrane of acinar cells; while calbindin-2, SERCA-2 and SERCA-3 were found inside the acinar cells; and PMCA-2 was found in the apical membrane and in the secretory canaliculi between the cells. Based on these findings, we propose the following model of calcium secretion in human salivary glands: (1) calcium enters the acinar cell at the basolateral side via calcium channel CaT-Like (calcium influx); (2) intracellular calcium is taken up into the endoplasmic reticulum by SERCA-2 and possibly SERCA3 or bound to calbindin-2 (intracellular calcium pool); and (3) calcium is secreted by PMCAs at the apical plasma membrane (calcium efflux). PMID- 16270204 TI - Modification of plant architecture through the expression of GA 2-oxidase under the control of an estrogen inducible promoter in Arabidopsis thaliana L. AB - The gibberellin (GA) 2-oxidase (PcGA2ox1) from bean catalyses the 2beta hydroxylation of some precursor and bioactive GAs resulting in their inactivation. We have expressed PcGA2ox1 under the control of the estrogen receptor-based chemical-inducible system, XVE, to modify plant architecture and assess whether transgene expression is localised. Applications of estradiol to the shoot apical region of inducible PcGA2ox1 overexpressors exhibited delays in both bolting (maximum of 46 days) and times to anthesis (maximum of 62 days) compared to wildtype (36 and 41 days, respectively), without altering leaf area. Individual treated leaves showed signs of epinasty and became dark green; such estradiol-treated regions maintained these 'green-islands' well beyond the onset of leaf senescence. Northern blots revealed that the PcGA2ox1 transcript could be detected within 1 h of treatment. The level of PcGA2ox1 transcript appeared to peak 3-5 h after estradiol application in both high and semi expressors. Quantitative Reverse Transcription (QRT)-PCR data showed that GA down-regulated genes AtGA3ox1, AtGA20ox1 and SCARECROW-LIKE3 (SCL3) were up-regulated and the GA up-regulated genes AtGA2ox1 and AtExp1 were down-regulated in estradiol-treated leaves of inducible PcGA2ox1 overexpressors; neighbouring non-treated leaves showing no significant changes. Further molecular analyses revealed that expression of the transgene was confined to estradiol-treated leaves only. Expression profiles of GA down- and up-regulated genes in inducer-treated overexpressors appeared to be synchronised with changes in leaf phenotype. These observations suggest that PcGA2ox1 under the control of the XVE system can be used effectively to alter plant architecture in Arabidopsis by localised 2beta hydroxylation of GAs at estradiol-treated sites. PMID- 16270205 TI - Genetic characterization of asymmetric somatic hybrids between Bupleurum scorzonerifolium Willd and Triticum aestivum L.: potential application to the study of the wheat genome. AB - In this paper, we describe how Bupleurum scorzonerifolium/Triticum aestivum asymmetric somatic hybrids can be exploited to study the wheat genome. Protoplasts of B. scorzonerifolium Willd were irradiated with ultraviolet light (UV) and fused with protoplasts of common wheat (T. aestivum L.). All cell clones were similar in appearance to those of B. scorzonerifolium, while the regenerated plantlets were either intermediate or B. scorzonerifolium-like. Genotypic screening using isozymes showed that 39.3% of cell clones formed were hybrid. Some of the hybrid cell clones grew vigorously, and differentiated green leaves, shoots or plantlets. DNA marker analysis of the hybrids demonstrated that wheat DNA was integrated into the nuclear genomes of B. scorzonerifolium and in situ karyotyping cells revealed that a few wheat chromosome fragments had been introgressed into B. scorzonerifolium. The average wheat SSR retention frequency of the RH panel was 20.50%, but was only 6.67% in fusions with a non-irradiated donor. B. scorzonerifolium chromosomes and wheat SSR fragments in most asymmetric hybrid cell lines remained stable over a period of 2.5-3.5 years. We suggest the UV-induced asymmetric somatic hybrids between B. scorzonerifolium Willd and T. aestivum L. have the potential for use in the construction of an RH map of the wheat genome. PMID- 16270206 TI - Trilateral retinoblastoma. PMID- 16270207 TI - [Synovial lesions around the knee joint]. AB - This article reviews the anatomy of synovial-lined structures around the knee joint including the joint capsule, plicae, bursae, and tendon sheaths. Pathologic conditions are grouped into either inflammatory, induced by deposition of various substances, or neoplastic. The MRI appearances of the various diseases are discussed. PMID- 16270208 TI - [Diagnostics for diseases of the gallbladder and biliary tract from the viewpoint of the internist and surgeon. Demands made on radiological diagnostics]. AB - Jaundice and colic pain of the right upper quadrant are the main symptoms of biliary diseases. Gallstone-related diseases often lead to hospital admission. The evaluation of a patient with biliary symptoms requires a combination of history taking, physical examination, laboratory analysis, and imaging modalities. A high-quality magnetic resonance imaging (MRI) or computed tomography (CT) scan is usually sufficient to evaluate a patient with painless jaundice. Ultrasonography is helpful as an initial screening test to guide the diagnostic work-up. Invasive methods (e.g., ERCP) are mainly used for palliation of patients with incurable disease. PMID- 16270209 TI - The "health benefit basket" in the Netherlands. AB - This contribution describes the entitlements in Dutch health care and explores how these entitlements are determined and to whom they apply. The focus is on services of curative care. No comprehensive positive or negative list of individual services is included in formal laws. Instead, the legislation states only what general types of medical services are covered and generally the "usual care" criterion determines to which interventions patients are entitled. This criterion is not very restrictive and yields local variations in service provision, which are moderated by practice guidelines. It is conceivable, however, that the recent introduction of the DBC financing system will change the reimbursement and therefore benefit-setting policy. PMID- 16270210 TI - Determining the "Health Benefit Basket" of the Statutory Health Insurance scheme in Germany: methodologies and criteria. AB - The issue of defining health benefit catalogues has recently gained new importance in Germany as a result of the creation of the new Institute for Quality and Efficiency. The Institute was designed to support the Federal Joint Committee conducting effectiveness studies for benefit coverage decisions. The Committee and the contractual partners (sickness funds and providers) define the benefit catalogues for the Statutory Health Insurance in the framework of Social Code Book V, Germany's most relevant health care scheme. Unlike other countries, the German federal government limits its regulatory role to defining procedures that determine the scope of Statutory Health Insurance services. The explicitness of the benefit catalogues varies greatly between different sectors. While benefits in outpatient care are rather explicitly defined, benefit definitions for inpatient care are vague. It is argued that the establishment of the new Institute and the development of the DRG system are initial steps towards a more effective and explicit benefit catalogue. PMID- 16270211 TI - The "Health Benefit Basket" in Denmark: a description of entitlements, actors, and decision-making processes in the curative health sector. AB - Until 2007, when the new legislation on health care becomes effective, the right to receive free health care services in Denmark, or "health benefits," are described in a comprehensive set of legislation, including laws, executive orders and legal guidelines. This contribution provides an overview of the current main legislation regulating the Danish "health benefit basket" and describes the regulatory mechanisms for the provision of curative care at Danish hospitals and primary health care offices. Although the services are both financed and planned by the counties, they differ substantially in the way that benefits are regulated. PMID- 16270212 TI - Defining the "Health Benefit Basket" in nine European countries. Evidence from the European Union Health BASKET Project. AB - This article identifies and analyses a framework for "health baskets," the taxonomy of benefit catalogues for curative services, and the criteria for the in or exclusion of benefits in nine EU member states (Denmark, England, France, Germany, Hungary, Italy, The Netherlands, Poland and Spain). Focusing on services of curative care, it is found that the explicitness of benefit catalogues varies largely between the countries. In the absence of explicitly defined benefit catalogues, in- and outpatient remuneration schemes have the character of benefit catalogues. The criteria for the in- or exclusion into benefit catalogues are often not transparent and (cost-)effectiveness is applied only for certain sectors. An EU-wide harmonization of benefit baskets does not seem realistic in the short or medium term as the variation in criteria and the taxonomies of benefit catalogues are large but not insurmountable. There may be scope for a European core basket. PMID- 16270213 TI - Results of a phase 2 study of valproic acid alone or in combination with all trans retinoic acid in 75 patients with myelodysplastic syndrome and relapsed or refractory acute myeloid leukemia. AB - Valproic acid (VPA) inhibits histone deacetylase activity and induces differentiation of acute myeloid leukemia (AML) blasts in vitro. We observed clinical responses to VPA in patients with myelodysplastic syndrome (MDS) and AML. Here, we report follow-up data on 75 patients. Of these, 66 were started on VPA monotherapy, with later addition of all-trans retinoic acid (ATRA) in patients who did not respond or relapsed. Nine patients were treated with VPA + ATRA from the start. Median treatment duration was 4 months for VPA and 2 months for ATRA. Hematological improvement, according to international working group criteria for MDS, was observed in 18 patients (24%). Median response duration was 4 months. ATRA exerted no additional effect in patients receiving the combination from the start or benefited primary VPA nonresponders. However, of ten VPA responders who relapsed, four achieved a second response after addition of ATRA. Response rates were strongly dependent on disease type according to WHO classification. We found a response rate of 52% in MDS patients with a normal blast count (refractory sideroblastic anemia, refractory cytopenia with multilineage dysplasia, and refractory sideroblastic cytopenia with multilineage dysplasia). The response rate was 6% in refractory anemia with excess blasts (I + II), 16% in AML, and 0% in chronic myelomonocytic leukemia. Bone marrow blast count was the only variable that predicted responses. We conclude that VPA is clinically useful in low-risk MDS. For patients with high-risk MDS, VPA may be combined with chemotherapy or demethylating drugs. If patients relapse after an initial response to VPA, ATRA has the potential to induce a prolonged second response. PMID- 16270214 TI - Is standardised (18)F-FDG uptake value an outcome predictor in patients with stage III non-small cell lung cancer? AB - PURPOSE: Recent studies have demonstrated the relevance of (18)F-FDG uptake as an independent prognostic factor for recurrence of operable non-small cell lung cancer (NSCLC). This corresponds with the experimental finding that FDG uptake correlates with the proliferative activity of tumour cells (Higashi et al., J Nucl Med 2000;41:85-92). On the basis of these observations, we studied the influence of FDG uptake on prognosis and occurrence of distant metastases in patients with advanced NSCLC. METHODS: One hundred and fifty-nine patients with NSCLC of UICC stage IIIA or IIIB were included in the study. In all patients, neoadjuvant treatment was planned to achieve operability. FDG PET was performed as an additional staging procedure prior to the initiation of therapy. Clinical outcome data in terms of overall survival, disease-free survival and incidence of distant metastases could be obtained for 137 patients and were correlated with the average standardised uptake value of the tumour (SUV(avg)). Furthermore, other factors influencing SUV(avg) and patient outcome (histological tumour type, grading, UICC stage, tumour size) were analysed. RESULTS: SUV(avg) was significantly influenced by tumour histology, UICC stage and tumour size. No significant difference could be shown for grading. In 38 out of the 159 patients (24%), FDG PET revealed previously unsuspected distant metastases. The incidence of distant metastases significantly correlated with SUV(avg). Overall survival tended to decrease with increasing SUV(avg); however, significance was only reached when a cut-off of 12.0 was applied (p=0.05). CONCLUSION: FDG uptake is an independent prognostic factor in patients with UICC stage III NSCLC, although less distinctively so than has been reported for stage I/II tumours. PMID- 16270216 TI - [Anonymous critical incident reporting system in anaesthesiology. Results after 18 months]. AB - Two years ago we implemented a reporting system for critical incidents in the Department of Anaesthesiology and Intensive Care of the University Hospital Dresden. During the first 18 months 162 anonymous reports were registered. The most common errors involved airway and ventilation management, followed by errors in fluid and cardio-vascular management. The main causes were distraction, lack of experience, specific training and communication deficits. The confidence in the anonymity of the reporting system was very high. Following the analysis of the reports, several modifications were initiated, e.g. specific training programs or definition of standards. Over time, a change in the relative distribution of reported errors was observed. The article discusses the different kinds of errors and possible countermeasures. It also strengthens several aspects which are important to consider during the initial phase of a local critical incident reporting system. PMID- 16270215 TI - [Use of the size 3 ProSeal laryngeal mask airway in children. Results of a randomized crossover investigation with the Classic laryngeal mask airway]. AB - BACKGROUND: The low-pressure airway seal of the Classic laryngeal mask airway (CLMA) can be inadequate for positive pressure ventilation (PPV) in children. The ProSeal laryngeal mask airway (PLMA) forms a more effective seal of the airway than the CLMA and facilitates gastric tube placement in adults. The size 3 PLMA can be used in adults and children. METHODS: The CLMA and PLMA were studied in random order -- crossover -- in 30 anaesthetized, non-paralysed children (average age 10.6 years, average body weight 39 kg). Airway leak pressure, maximum tidal volume, ease of insertion, quality of initial airway and fiberoptic position were determined. Gastric tube placement was assessed for the PLMA. RESULTS: The mean airway leak pressure in neutral head position (27.0 vs. 16.8 cm H(2)O), maximum flexion (38.3 vs. 26.2 cm H(2)O) and maximum extension (21.1 vs. 14.2 cm H(2)O) as well as the mean maximum tidal volume (1432 vs. 1062 ml) were significantly higher (p<0.001) for the PLMA. Air insufflation into the stomach occurred with the CLMA but not with the PLMA. Gastric tube placement was possible in all patients. CONCLUSIONS: The high reliability of g-tube placement and the significantly increased airway leak pressure seem to make the size 3 PLMA a more suitable device for PPV in children than the same size CLMA. PMID- 16270217 TI - Complex microcolinearity among wheat, rice, and barley revealed by fine mapping of the genomic region harboring a major QTL for resistance to Fusarium head blight in wheat. AB - A major quantitative trait locus (QTL), Qfhs.ndsu-3BS, for resistance to Fusarium head blight (FHB) in wheat has been identified and verified by several research groups. The objectives of this study were to construct a fine genetic map of this QTL region and to examine microcolinearity in the QTL region among wheat, rice, and barley. Two simple sequence repeat (SSR) markers (Xgwm533 and Xgwm493) flanking this QTL were used to screen for recombinants in a population of 3,156 plants derived from a single F(7) plant heterozygous for the Qfhs.ndsu-3BS region. A total of 382 recombinants were identified, and they were genotyped with two more SSR markers and eight sequence-tagged site (STS) markers. A fine genetic map of the Qfhs.ndsu-3BS region was constructed and spanned 6.3 cM. Based on replicated evaluations of homozygous recombinant lines for Type II FHB resistance, Qfhs.ndsu-3BS, redesignated as Fhb1, was placed into a 1.2-cM marker interval flanked by STS3B-189 and STS3B-206. Primers of STS markers were designed from wheat expressed sequence tags homologous to each of six barley genes expected to be located near this QTL region. A comparison of the wheat fine genetic map and physical maps of rice and barley revealed inversions and insertions/deletions. This suggests a complex microcolinearity among wheat, rice, and barley in this QTL region. PMID- 16270219 TI - Isolation and characterization of 18 genes encoding alpha- and beta-expansins in wheat (Triticum aestivum L.). AB - Expansins are thought to be key regulators of cell wall extension during plant growth. In this study, we isolated 18 expansin genes from wheat, nine of which encode alpha-expansins while the other nine code for beta-expansins. The cysteine rich and tryptophan-rich regions of the deduced amino acid sequences of all 18 expansins were highly conserved. Genomic sequences were obtained for 17 of the genes, and their intron patterns were determined. Four (A, C, D, E) of the six intron positions known in expansin genes from other species were found to be occupied in these wheat expansin genes. Five wheat expansin genes were mapped to chromosomes 1L, 2L, 5L and 6L respectively, by in silico and comparative mapping. The 18 wheat expansin genes were expressed in leaf, root and the developing seed. Moreover, it was demonstrated that four beta-expansin genes were up-regulated in the internode tissue in F1 hybrids, suggesting that changes in the regulation of these genes in hybrid might contribute to the heterosis observed in internode length and plant height. We therefore conclude that expansins are encoded by a multigene family in wheat, and could play important roles in growth and development. PMID- 16270218 TI - Kinetochore fiber formation in animal somatic cells: dueling mechanisms come to a draw. AB - The attachment to and movement of a chromosome on the mitotic spindle are mediated by the formation of a bundle of microtubules (MTs) that tethers the kinetochore on the chromosome to a spindle pole. The origin of these "kinetochore fibers" (K fibers) has been investigated for over 125 years. As noted in 1944 by Schrader [Mitosis, Columbia University Press, New York, 110 pp.], there are three possible ways to form a K fiber: (a) it grows from the pole until it contacts the kinetochore, (b) it grows directly from the kinetochore, or (c) it forms as a result of an interaction between the pole and the chromosome. Since Schrader's time, it has been firmly established that K fibers in centrosome-containing animal somatic cells form as kinetochores capture MTs growing from the spindle pole (route a). It is now similarly clear that in cells lacking centrosomes, including higher plants and many animal oocytes, K fibers "self-assemble" from MTs generated by the chromosomes (route b). Can animal somatic cells form K fibers in the absence of centrosomes by the "self-assembly" pathway? In 2000, the answer to this question was shown to be a resounding "yes." With this result, the next question became whether the presence of a centrosome normally suppresses K fiber self-assembly or if this route works concurrently with centrosome-mediated K-fiber formation. This question, too, has recently been answered: observations on untreated live animal cells expressing green fluorescent protein-tagged tubulin clearly show that kinetochores can nucleate the formation of their associated MTs in a unique manner in the presence of functional centrosomes. The concurrent operation of these two "dueling" routes for forming K fibers in animal cells helps explain why the attachment of kinetochores and the maturation of K fibers occur as quickly as they do on all chromosomes within a cell. PMID- 16270220 TI - Factors affecting histological regression of crescentic Henoch-Schonlein nephritis in children. AB - To identify the factors affecting histological regression of crescentic Henoch Schonlein nephritis (HSN), we retrospectively analyzed serially biopsied 20 children with crescentic HSN treated with immunosuppressants. They were classified into two groups according to the histological changes between the first and second biopsy: group I (n=10) with histological regression and group II (n=10) with no change or histological progression. Of the 20 patients, 19 showed a favorable outcome at the end of follow-up. Initial laboratory and histological findings did not differ between the two groups. Histological regression was associated with a younger age at onset (P=0.003), early treatment with immunosuppressants (P=0.044) and absent or decreased fibrinogen deposits at the second biopsy (P<0.0001) in a univariate analysis. Mesangial IgA and fibrinogen depositions at the second biopsy were reduced significantly in group I (P<0.05). In the multivariate analysis, a younger age was an independent determinant of histological regression (OR 1.44; 95% CI 1.03-2.02). The intensity of fibrinogen deposits at the second biopsy correlated positively with the age at onset (r=0.503, P=0.024), and the chronicity index at the second biopsy correlated positively with the time that immunosuppressive therapy was started (r=0.619, P=0.004). PMID- 16270221 TI - Long-term outcome of chronic dialysis in children. AB - We describe the outcome since 1984 of all children receiving chronic dialysis in our centre for >3 months with a minimum follow-up of 5 (median 7.2) years. There were 98 children (61 boys), with a median age at the start of dialysis of 4.2 (range: birth to 16.2) years. Twenty-one children started dialysis at <1 year of age and 54 under <5 years. Thirty children had significant comorbidity. The median time on dialysis was 1.4 (0.3 to 14.4) years, giving a total dialysis experience of 296 patient-years. Fifty-three children received a renal transplant as their first change of treatment modality, but 31 switched between PD and HD, with a total of 54 changes of dialysis modality pre-transplantation. Twenty-one of the transplanted patients (39%) returned to dialysis. There were a total of 115 transplants in 88 patients. There was a positive increase for both the weight and height SDS for all the age groups while on dialysis, but this did not reach statistical significance. There were 17 deaths over the 20-year study period; of these, 10 died on dialysis. The overall patient survival was 83%. The mortality rate was 2.7 times greater in children who required renal replacement therapy under the age of 5 years. Of the deaths, 76% were in association with comorbid conditions. In conclusion, both a younger age at the start of renal replacement therapy and comorbidity are significant risk factors for death. The number of returnees to dialysis highlights the importance of conserving dialysis access. PMID- 16270222 TI - A randomized prospective controlled trial of antibiotic prophylaxis in intraoral bone grafting procedures: single-dose clindamycin versus 24-hour clindamycin prophylaxis. AB - PURPOSE: The objective of this study was to compare the efficacy of a single oral dose of clindamycin with a 24-h protocol of clindamycin administration in local buccal onlay grafting procedures. MATERIAL AND METHODS: A prospective randomized study in 124 patients was performed. Eligible patients were randomized to receive antibiotic prophylaxis either as a single dose (group I) or over a 24-h period (group II). In both groups prophylaxis started with an oral dose of 600 mg clindamycin 1 h before surgery, followed by either placebo or 300 mg clindamycin every 6 h. The primary endpoint of this study was wound infection at the receptor site within 8 weeks after surgery. Secondary outcome measurements included postoperative infections at the donor site and adverse events as a result of antibiotic administration. RESULTS: The mean age of the patients was 35.9+/-10.1 years (range 18-59 years). Infections at the receptor site were seen in two patients (3.2%, 95% CI 0-7.6%) of the single-dose group and in three patients (4.8%, 95% CI 0-10.1%) of the 24-h group. Infection at the donor site occurred in four patients (6.4%, 95% CI 0-12.5%) of the single-dose group and in two patients (3.2%, 95% CI 0-7.6%) of the 24-h group. Postoperative infections were predominantly caused by alpha-hemolytic streptococci sensitive to penicillin. CONCLUSIONS: No statistically significant difference was found between the prophylactic single dose of clindamycin and the 24-h regimen of clindamycin with regard to postoperative infection in patients undergoing local bone augmentation procedures. PMID- 16270223 TI - Left ventricular apical ballooning. AB - Transient left ventricular apical ballooning is an acute cardiac syndrome characterised by an abrupt onset of chest pain and is usually associated with anterior electrocardiographic changes. The observed apical ballooning of the left ventricle is usually reversible and develops in the absence of significant atherosclerotic coronary artery disease. Here we present (to our knowledge) the first case of this syndrome in a negroid patient. PMID- 16270224 TI - Controlled expression of recombinant proteins in Physcomitrella patens by a conditional heat-shock promoter: a tool for plant research and biotechnology. AB - The ability to express tightly controlled amounts of endogenous and recombinant proteins in plant cells is an essential tool for research and biotechnology. Here, the inducibility of the soybean heat-shock Gmhsp17.3B promoter was addressed in the moss Physcomitrella patens, using beta-glucuronidase (GUS) and an F-actin marker (GFP-talin) as reporter proteins. In stably transformed moss lines, Gmhsp17.3B-driven GUS expression was extremely low at 25 degrees C. In contrast, a short non-damaging heat-treatment at 38 degrees C rapidly induced reporter expression over three orders of magnitude, enabling GUS accumulation and the labelling of F-actin cytoskeleton in all cell types and tissues. Induction levels were tightly proportional to the temperature and duration of the heat treatment, allowing fine-tuning of protein expression. Repeated heating/cooling cycles led to the massive GUS accumulation, up to 2.3% of the total soluble proteins. The anti-inflammatory drug acetyl salicylic acid (ASA) and the membrane fluidiser benzyl alcohol (BA) also induced GUS expression at 25 degrees C, allowing the production of recombinant proteins without heat-treatment. The Gmhsp17.3B promoter thus provides a reliable versatile conditional promoter for the controlled expression of recombinant proteins in the moss P. patens. PMID- 16270225 TI - ZmMPK6, a novel maize MAP kinase that interacts with 14-3-3 proteins. AB - Although an increasing body of evidence indicates that plant MAP kinases are involved in a number of cellular processes, such as cell cycle regulation and cellular response to abiotic stresses, hormones and pathogen attack, very little is known about their biochemical properties and regulation mechanism. In this paper we report on the identification and characterization of a novel member of the MAP kinase family from maize, ZmMPK6. The amino acid sequence reveals a high degree of identity with group D plant MAP kinases. Recombinant ZmMPK6, expressed in Escherichia coli, is an active enzyme able to autophosphorylate. Remarkably, ZmMPK6 interacts in vitro with GF14-6, a maize 14-3-3 protein and the interaction is dependent on autophosphorylation. The interacting domain of ZmMPK6 is on the C terminus and is comprised between amino acid 337 and amino acid 467. Our results represent the first evidence of an interaction between a plant MAP kinase and a 14-3-3 protein. Possible functional roles of this association in vivo are discussed. PMID- 16270227 TI - DcMYB1 acts as a transcriptional activator of the carrot phenylalanine ammonia lyase gene (DcPAL1) in response to elicitor treatment, UV-B irradiation and the dilution effect. AB - Expression of a carrot phenylalanine ammonia-lyase (PAL) gene (DcPAL1) in suspension-cultured carrot cells is induced by treatment with a fungal elicitor, ultraviolet B (UV-B) irradiation, and by transferring and diluting cells with fresh medium (the dilution effect). Box-L-like sequences are known as important cis-elements of genes for enzymes involved in the phenylpropanoid biosynthetic pathway. Six sequences, box-L0 to box-L5, exist in the DcPAL1 gene promoter region. In this study, we isolated cDNA encoding the R2R3 type of MYB transcription factor, DcMYB1, using yeast one-hybrid screening with box-L1 or box L5 as target elements. DcMYB1 bound to boxes-L0, L1, L3/4, and L5 sequences (ACC(A/T)(A/T)CC) in vitro, and in yeast cells and carrot protoplasts. Transient expression of DcMYB1 could up-regulate DcPAL1 promoter activity in carrot protoplasts. Results of the transient expression experiment for the deletion mutated promoters of boxes-L0, L1, L3, and L5 suggest that these box-L-like sequences were required for the complete activation of the DcPAL1 promoter by DcMYB1. Expression of DcMYB1 transcripts was induced 0.5 h after elicitor treatment or UV-B irradiation, and 2 h after the dilution effect. Induction of DcPAL1 expression occurred 1 h after DcMYB1 expression in all stress treatments, and repression of DcMYB1 expression by RNA interference caused cessation of the up-regulation of DcPAL1 expression in the elicitor treatment or with UV-B irradiation. These results suggest that DcMYB1 is the main regulatory factor acting on box-L sequences in the DcPAL1 gene that respond to environmental cues. PMID- 16270226 TI - The evolution of the abscisic acid-response in land plants: comparative analysis of group 1 LEA gene expression in moss and cereals. AB - The moss Physcomitrella patens possesses a single copy of a Group 1 LEA gene, designated PpLEA-1. Sequence analysis of the PpLEA-1 gene reveals the gene to contain a single intron in a position conserved in all members of the Group 1 LEA gene family, but also to contain a premature termination codon within the first exon. Nevertheless, a PpLEA-1 transcript accumulates in moss tissues in response both to the imposition of osmotic stress, and to the plant growth regulator abscisic acid (ABA). This response appears to be mediated at the transcriptional level, and observation of the pattern of gene expression, reported by histochemical staining of plants expressing a PpLea-1::GUS transgene suggests that the promoter responds preferentially to ABA in protonemal filaments, whereas osmotic stress induces gene expression primarily in the gametophores. Quantitative analysis of promoter activity by transient expression in Physcomitrella protoplasts shows the PpLEA-1 promoter to be highly active in response to ABA and osmotic stress. ABA-mediated transgene expression from the PpLea-1 promoter occurs at a level similar to that driven by the highly active promoter of the wheat Group 1 LEA gene, E(m). Site-directed mutagenesis of the PpLEA-1 promoter indicates that ABA-inducibility is mediated via an ACGT-core motif similar to that seen in the ABA response elements of higher plant LEA genes. However, whereas the wheat E(m )promoter is active in moss tissues, the moss promoter is not reciprocally active in cereal cells: no activity, ABA inducible or otherwise was detected in barley aleurone protoplasts transfected with the PpLEA-1::GUS construct. We propose that ABA activation of gene expression in moss cells represents an ancestral state, with only minimal requirements for promoter recognition, whereas cereal cells require the interaction of additional factors with ABA-responsive promoters. PMID- 16270229 TI - Expression of the hypersensitive response-assisting protein in Arabidopsis results in harpin-dependent hypersensitive cell death in response to Erwinia carotovora. AB - Active defense mechanisms of plants against pathogens often include a rapid plant cell death known as the hypersensitive cell death (HCD). Hypersensitive response assisting protein (HRAP) isolated from sweet pepper intensifies the harpin(Pss) mediated HCD. Here we demonstrate that constitutive expression of the hrap gene in Arabidopsis results in an enhanced disease resistance towards soft rot pathogen, E. carotovora subsp. carotovora. This resistance was due to the induction of HCD since different HCD markers viz. Athsr3, Athsr4, ion leakage, H(2)O(2) and protein kinase were induced. One of the elicitor harpin proteins, HrpN, from Erwinia carotovora subsp. carotovora was able to induce a stronger HCD in hrap-Arabidopsis than non-transgenic controls. To elucidate the role of HrpN, we used E. carotovora subsp. carotovora defective in HrpN production. The hrpN(-) mutant did not induce disease resistance or HCD markers in hrap-Arabidopsis. These results imply that the disease resistance of hrap-Arabidopsis against a virulent pathogen is harpin dependent. PMID- 16270228 TI - Metabolite profiling reveals a role for atypical cinnamyl alcohol dehydrogenase CAD1 in the synthesis of coniferyl alcohol in tobacco xylem. AB - In angiosperms, lignin is built from two main monomers, coniferyl and sinapyl alcohol, which are incorporated respectively as G and S units in the polymer. The last step of their synthesis has so far been considered to be performed by a family of dimeric cinnamyl alcohol dehydrogenases (CAD2). However, previous studies on Eucalyptus gunnii xylem showed the presence of an additional, structurally unrelated, monomeric CAD form named CAD1. This form reduces coniferaldehyde to coniferyl alcohol, but is inactive on sinapaldehyde. In this paper, we report the functional characterization of CAD1 in tobacco (Nicotiana tabacum L.). Transgenic tobacco plants with reduced CAD1 expression were obtained through an RNAi strategy. These plants displayed normal growth and development, and detailed biochemical studies were needed to reveal a role for CAD1. Lignin analyses showed that CAD1 down-regulation does not affect Klason lignin content, and has a moderate impact on G unit content of the non-condensed lignin fraction. However, comparative metabolic profiling of the methanol-soluble phenolic fraction from basal xylem revealed significant differences between CAD1 down regulated and wild-type plants. Eight compounds were less abundant in CAD1 down regulated lines, five of which were identified as dimers or trimers of monolignols, each containing at least one moiety derived from coniferyl alcohol. In addition, 3-trans-caffeoyl quinic acid accumulated in the transgenic plants. Together, our results support a significant contribution of CAD1 to the synthesis of coniferyl alcohol in planta, along with the previously characterized CAD2 enzymes. PMID- 16270230 TI - Identification and characterisation of the alpha and beta subunits of succinyl CoA ligase of tomato. AB - Despite the central importance of the TCA cycle in plant metabolism not all of the genes encoding its constituent enzymes have been functionally identified. In yeast, the heterodimeric protein succinyl CoA ligase is encoded for by two single copy genes. Here we report the isolation of two tomato cDNAs coding for alpha- and one coding for the beta-subunit of succinyl CoA ligase. These three cDNAs were used to complement the respective Saccharomyces cerevisiae mutants deficient in the alpha- and beta-subunit, demonstrating that they encode functionally active polypeptides. The genes encoding for the subunits were expressed in all tissues, but most strongly in floral and leaf tissues, with equivalent expression of the two alpha-subunit genes being expressed to equivalent levels in all tissues. In all instances GFP fusion expression studies confirmed an expected mitochondrial location of the proteins encoded. Following the development of a novel assay to measure succinyl CoA ligase activity, in the direction of succinate formation, the evaluation of the maximal catalytic activities of the enzyme in a range of tissues revealed that these paralleled those of mRNA levels. We also utilized this assay to perform a preliminary characterisation of the regulatory properties of the enzyme suggesting allosteric control of this enzyme which may regulate flux through the TCA cycle in a manner consistent with its position therein. PMID- 16270231 TI - Hairy root-activation tagging: a high-throughput system for activation tagging in transformed hairy roots. AB - Activation tagging is a powerful technique for generating gain-of-function mutants in plants. We developed a new vector system for activation tagging of genes in "transformed hairy roots". The binary vector pHR-AT (Hairy Root Activation Tagging) and its derivative pHR-AT-GFP contain a cluster of rol (rooting locus) genes together with the right border facing four tandem repeats of the cauliflower mosaic virus (CaMV) 35S enhancer element on the same T-DNA. Transformation experiments using Arabidopsis, potato, and tobacco as model plants revealed that upon inoculating plants with Agrobacterium tumefaciens harboring these vectors, a large number of independently transformed roots could be induced from explants within a short period of time, and root culture lines were subsequently established. Molecular analyses of the pHR-AT-GFP-transformed Arabidopsis lines showed that expression of the genes adjacent to the T-DNA insertion site was significantly increased. This system may facilitate application of the activation-tagging approach to plant species that are recalcitrant to the regeneration of transgenic plants. High-throughput metabolic profiling of activation-tagged root culture lines will offer opportunities for identifying regulatory or biosynthetic genes for the production of valuable secondary metabolites of interest. PMID- 16270232 TI - A putative plasma membrane cation/proton antiporter from soybean confers salt tolerance in Arabidopsis. AB - Cation transport is thought to be an important process for ion homeostasis in plant cells. Here, we report that a soybean putative cation/proton antiporter GmCAX1 may be a mediator of this process. GmCAX1 is expressed in all tissues of the soybean plants but at a lower level in roots. Its expression was induced by PEG, ABA, Ca(2+), Na(+) and Li(+) treatments. The GmCAX1-GFP fusion protein was mainly localized in plasma membrane of the transgenic Arabidopsis plant cells and onion epidermal cells. Transgenic Arabidopsis plants overexpressing GmCAX1 accumulated less Na(+), K(+), and Li(+), and were more tolerant to elevated Li(+) and Na(+) levels during germination when compared with the controls. These results suggest that GmCAX1 may function as an antiporter for Na(+), K(+) and Li(+). Modulation of this antiporter may be beneficial for regulation of ion homeostasis and thus plant salt tolerance. PMID- 16270233 TI - Dual DNA binding property of ABA insensitive 3 like factors targeted to promoters responsive to ABA and auxin. AB - The ABA responsive ABI3 and the auxin responsive ARF family of transcription factors bind the CATGCATG (Sph) and TGTCTC core motifs in ABA and auxin response elements (ABRE and AuxRE), respectively. Several evidences indicate ABI3s to act downstream to auxin too. Because DNA binding domain of ABI3s shows significant overlap with ARFs we enquired whether auxin responsiveness through ABI3s could be mediated by their binding to canonical AuxREs. Investigations were undertaken through in vitro gel mobility shift assays (GMSA) using the DNA binding domain B3 of PvAlf (Phaseolus vulgaris ABI3 like factor) and upstream regions of auxin responsive gene GH3 (-267 to -141) and ABA responsive gene Em (-316 to -146) harboring AuxRE and ABRE, respectively. We demonstrate that B3 domain of PvAlf could bind AuxRE only when B3 was associated with its flanking domain B2 (B2B3). Such strict requirement of B2 domain was not observed with ABRE, where B3 could bind with or without being associated with B2. This dual specificity in DNA binding of ABI3s was also demonstrated with nuclear extracts of cultured cells of Arachis hypogea. Supershift analysis of ABRE and AuxRE bound nuclear proteins with antibodies raised against B2B3 domains of PvAlf revealed that ABI3 associated complexes were detectable in association with both cis elements. Competition GMSA confirmed the same complexes to bind ABRE and AuxRE. This dual specificity of ABI3 like factors in DNA binding targeted to natural promoters responsive to ABA and auxin suggests them to have a potential role in conferring crosstalk between these two phytohormones. PMID- 16270234 TI - Effects of stimulation mode, level and location on forward-masked excitation patterns in cochlear implant patients. AB - In multi-channel cochlear implants, electrical current is delivered to appropriate electrodes in the cochlea to approximate the spatial representation of speech. Theoretically, electrode configurations that restrict the current spread within the cochlea (e.g., bi- or tri-polar stimulation) may provide better spatial selectivity, and in turn, better speech recognition than configurations that produce a broader current spread (e.g., monopolar stimulation). However, the effects of electrode configuration on supra-threshold excitation patterns have not been systematically studied in cochlear implant patients. In the present study, forward-masked excitation patterns were measured in cochlear implant patients as functions of stimulation mode, level and location within the cochlea. All stimuli were 500 pulses-per-second biphasic pulse trains (200 micros/phase, 20 micros inter-phase gap). Masker stimuli were 200 ms in duration; the bi-polar configuration was varied from narrow (BP+1) to wide (BP+17), depending on the test condition. Probe stimuli were 20 ms in duration and the masker-probe delay was 5 ms; the probe configuration was fixed at BP+1. The results indicated that as the distance between the active and return electrodes in a bi-polar pair was increased, the excitation pattern broadened within the cochlea. When the distance between active and return electrodes was sufficiently wide, two peaks were often observed in the excitation pattern, comparable to non-overlapping electric fields produced by widely separated dipoles. Analyses of the normalized data showed little effect of stimulation level on the shape of the excitation pattern. PMID- 16270236 TI - Six characteristics of effective structured reporting and the inevitable integration with speech recognition. AB - The reporting of radiological images is undergoing dramatic changes due to the introduction of two new technologies: structured reporting and speech recognition. Each technology has its own unique advantages. The highly organized content of structured reporting facilitates data mining and billing, whereas speech recognition offers a natural succession from the traditional dictation transcription process. This article clarifies the distinction between the process and outcome of structured reporting, describes fundamental requirements for any effective structured reporting system, and describes the potential development of a novel, easy-to-use, customizable structured reporting system that incorporates speech recognition. This system should have all the advantages derived from structured reporting, accommodate a wide variety of user needs, and incorporate speech recognition as a natural component and extension of the overall reporting process. PMID- 16270237 TI - [Graz meeting on psychoses -- approaches to the etiology of schizophrenic disorders]. PMID- 16270235 TI - Comparison of positron emission tomography, computed tomography, and endoscopic ultrasound in the initial staging of patients with esophageal cancer. AB - INTRODUCTION: Improvement in esophageal cancer staging is needed. Positron emission tomography (PET), computed tomography (CT), and endoscopic ultrasound (EUS) in the staging of esophageal carcinoma were compared. METHODS: PET, CT, and EUS were performed and interpreted prospectively in 75 patients with newly diagnosed esophageal cancer. Either tissue confirmation or fine needle aspiration (FNA) was used as the gold standard of disease. Sensitivity and specificity for tumor, nodal, and metastatic (TNM) disease for each test were determined. TNM categorizations from each test were used to assign patients to subgroups corresponding to the three treatment plans that patients could theoretically receive, and these were then compared. RESULTS: Local tumor staging (T) was done correctly by CT and PET in 42% and by EUS in 71% of patients (P value > 0.14). The sensitivity and specificity for nodal involvement (N) by modality were 84% and 67% for CT, 86% and 67% for EUS, and 82% and 60% for PET (P value > 0.38). The sensitivity and specificity for distant metastasis were 81% and 82% for CT, 73% and 86% for EUS, and 81% and 91% for PET (P value > 0.25). Treatment assignment was done correctly by CT in 65%, by EUS in 75%, and by PET in 70% of patients (P value > 0.34). CONCLUSIONS: EUS had superior T staging ability over PET and CT in our study group. The tests showed similar performance in nodal staging and there was a trend toward improved distant disease staging with CT or PET over EUS. Assignment to treatment groups in relation to TNM staging tended to be better by EUS. Each test contributed unique patient staging information on an individual basis. PMID- 16270238 TI - [Etiopathogenetic theories in historical concepts of schizophrenia]. AB - In the years between 1800 and 1914, the real beginning of the twentieth century, different etiological theories and models were developed about illnesses which we today know as schizophrenic disorders. Particularly in the German speaking world, in the first third of the nineteenth century many of these followed a religious metaphysical paradigm. Those who paved the way for modern psychiatry represented an etiological concept with varying emphasis on brain pathological, hereditary degenerative or psycho-dynamic as well as sociogenic perspectives. It has become apparent that psychiatry was always in danger of giving way to reductionist approaches. Every reductionism emphasises merely partial truths. The systemic perspective teaches us that many points of view do not contradict each other but rather compliment each other. PMID- 16270239 TI - [Consequences of nosographic and trans-nosological concepts of schizophrenia]. AB - This essay starts with a brief remark about the -- necessary -- de-tours of the historical pathway to the concept of schizophrenia. The first part on nosographic approaches then describes the results of cross-sectional and longitudinal psychopathological symptom-assembling, subgrouping by factorial analyses, and the cross-cultural stability of the emerging disease concept: The human idiopathic syndrome of structural mental decline in its clinical variegation. The second part about de-nosological approaches provides an account of the clinical concepts of basic symptoms, spectrum disorders, neuropsychological deficits and their genetic family loading. Genetics of neuropsychological deficits demonstrate in detail that risk and protective factors to schizophrenia are genetically closely enmeshed. Higher familial loading of some neuropsychological deficits compared to the actual disease itself points at the dilemma that functional psychopathology deepens causal understanding and yet looses specificity for the disease syndrome. The concluding part of this essay emphasizes some sequelae of de-nosological approaches: Better tools for earlier recognition treatment and prevention; ego distal concepts of basic dysfunctions rather than the previous ego-proximal concepts, hence better self-esteem, insight, and de-stigmatization for patients. PMID- 16270240 TI - [The history of schizophrenias: philosophical roots]. AB - Although the term "schizophrenia" has been introduced in medical usage by E. Bleuler, all variations of schizophrenia developed afterwards trace back to E. Kraepelin. In his work on dementia praecox he intended to discover a yet unknown entity from nature definitely following the principles and maxima of positivism. In the following a great number of different concepts of schizophrenia were developed and the psychiatrist of the seventies and eighties of the last century was left confronted with an abundance of varying schizophrenic criteria. The upcoming globalization in diagnostic stopped this process of diversity and the ideas of Kraepelin reappeared in the ICD-10 criteria for schizophrenia with only a few modifications. The main problem of positivistic research approaches is that nature obviously is completely unimpressed by human made principles of rules and systems. Nature itself does not know these forms and categories invented by human beings. That is one of the reasons why positivistic schizophrenic research considering human made categories as natural has not been quite successful. A possible way out of this diagnostic dilemma -- insufficient categorical classification systems on the one hand and the necessity of apprehending psychopathological phenomena for an effective therapy on the other -- can be a change of paradigms from the usual categorical diagnostics based on the maxima of positivism to dimensional diagnostics developed in the frame of post-modern strategies of thinking. Such process-oriented diagnostic approaches considering the singular phenomenon as well as its significance for the individual and its pathogenesis as main foci of diagnostics provide the possibility for post-modern psychiatrists to start a new dialogue overcoming a positivism based monologue made by experts on the sufferers. PMID- 16270242 TI - [Pathophysiology of schizophrenia and its impact on pharmacotherapy]. AB - Schizophrenia is a severe psychiatric illness with a lifetime morbidity risk of around 1 %. Symptoms include hallucinations, delusions, poverty of thought and emotion and social withdrawal and cognitive deficits. Although newer antipsychotics affecting multiple neurotransmitter receptors facilitate therapy, many patients still do not achieve full response. Despite intensive study, the molecular etiology of schizophrenia remains enigmatic in many ways. The dopamine hypothesis of schizophrenia still plays an important role, although pharmacological studies, brain imaging analyses and genetic research indicate additional dysfunctions of glutamate, GABA and serotonin transmission. This article reviews the pathophysiological background of the disorder, its implications for pharmacological treatment and possible directions for future research. PMID- 16270241 TI - [Schizophrenia -- an existential disease]. AB - This paper deals with familiar findings in the field of schizophrenic psychosis re-interpreted from a medical anthropological (V. Weizsacker) as well as theoretical pathological (W. Dorr) view-point. Schizophrenic psychosis is thus discussed as an existential illness of the person as well as an aesthetic challenge for a certain cultural framework. This paper therefore investigates phenomenologically three main points that are not new but ought to be continuously re-considered: 1. The cultural-scientific thesis of a uniform onto- and pathogeny: Schizophrenic psychosis serves as topical example for the genetic and ontogenetic uniformity of so-called organic and psychic illnesses. 2. The anthropological thesis of the existential "Spiegelfunktion" of schizophrenic psychosis: From the phenomenological point of view, schizophrenic psychosis is defined as an existential illness hitting and reflecting the primary nature of human beings in a shocking way. 3. The therapeutic thesis of the effect and scientific basis of the paradoxical intervention: This thesis aims at describing the impossibility of understanding schizophrenic psychosis as a pathological but rather an anthropological phenomenon and problem of life, the acknowledgment of which shows therapeutic effect. The article's main concern is to discuss schizophrenic psychosis as phenomenon of existence and suffering, however not understandable, yet reflective of the existential predicament of humanity. This cultural-scientific concern is considered a permanent aim of subject-oriented medical analysis. PMID- 16270243 TI - [Biological hypotheses of schizophrenia: possible influences of immunology and endocrinology]. AB - A great number of studies show biological alterations in patients with schizophrenia, but many of these data are conflicting. Schizophrenia is a vastly heterogeneous disorder, most likely not caused by one etiological factor, but rather due to a complex network of different, interacting pathogenic influences. Variable clinical pictures may reflect different etiological factors. In a comprehensive theory of the origin of schizophrenic disorders, genetic and environmental influences cause changes in neuronal development which result in functional alterations of different neurotransmitter systems. Immunological research in schizophrenia was initially based on the "infection hypothesis" which was triggered by observing schizophrenia-like psychoses after influenza pandemic. Numerous immunological studies focusing on antibodies against specific viruses, unspecific antibodies and different other immune-phenomena were carried out in schizophrenia patients. Although the variability of the results from these studies is strikingly high, subgroups of patients with schizophrenia show an activated inflammatory response system with increased levels of proinflammatory cytokines and acute phase proteins. Furthermore, some investigations find changing activities in the T-cell system with a shift of TH-1 to an increased TH 2 activity. Endocrinological factors which may play a relevant role in the etiopathogenesis of schizophrenia include sex hormones and all changes caused by stress or other influences which are directly related to the HPA-axis. Alterations of the immune and the endocrinological systems might be caused by environmental factors like infections or exogenous stress. Due to the intensive interaction between the central nervous system, the immune system and different hormones the "development of a pathology" like schizophrenia can be seen in an integrative but multifactorial fashion. The clinical manifestation, the severity and the course of the disease might then be modulated by genetic vulnerability, the time of the "primary insult" -- which could be an infection, or psychological stress -- and its neuronal localisation and intensity. Different compensatory and decompensatory mechanisms in later life very likely play a crucial role for the further course of the disorder. PMID- 16270244 TI - [Genetic risk factors in schizophrenia]. AB - The high pathogenetic relevance of genetic factors in schizophrenia is beyond doubt based on the findings of epidemiological studies. By means of a complex mode of transmission, it is likely that several genes with weak to moderate effect jointly constitute a genetic basis for a vulnerability to schizophrenia that may well vary for different individuals. Other organic and psychosocial factors also play an individually different -- in some cases significant -- role in terms of pathogenesis, as a result of which an oligogenic/polygenic multifactor model is assumed from the standpoint of aetiopathogenetics. Molecular genetic methods consist in linkage analyses and association analyses. Positive linkage findings accumulate particularly for the chromosomes 1q, 6p, 8p, 13q and 22q. By themselves, individual mutations contribute little to the range of schizophrenic feature characteristics, it was not possible -- irrespective of some subtypes -- to replicate genes of major effect. From the large number of possible candidate genes, although studies on DRD3, DRD2 and HTR2A produced positive results, the magnitudes of effect were low. The findings for alleles of dysbindin, neuregulin 1, DAO, COMT, PRODH, ZDHHC and DISC are less clear. The search for schizophrenia-relevant mutations is hampered by the possibility of a heterogeneous phenotype of schizophrenia in case of a homogeneous genotype as much as by the possibility of inter-individually homogeneous phenotypical characteristics in case of schizophrenia-relevant heterotype in the genome. With the aid of the concept of endo-phenotypes, based on neurobiological phenomena, it might be possible to take a more direct approach that leads from relevant mutations to the risk of schizophrenias. However, replacing schizophrenic alienation with neurobiological aspects leads to difficulties in explaining these complex disorder profiles. Schizophrenic diseases require an explanatory approach that also incorporates personality and developmental psychological aspects from the outset, if the aim is not to restrict type of schizophrenic disease exclusively to loci of molecular genetic changes. PMID- 16270245 TI - [Neuroimaging in schizophrenia]. AB - This overview is focused on functional neuroimaging including functional magnetic resonance imaging (fMRI), positron emission tomography (PET) and single photon emission computed tomography (SPECT). Recent evidence for the "dopamine hypothesis of schizophrenia" is summarized including alterations of presynaptic dopamine metabolism and postsynaptic receptor binding potential. Emphasis is given to dopaminergic challenge studies using amphetamine and AMPT. Several PET and SPECT studies have shown a pronounced increase of amphetamine-induced dopamine release as well as decrease of AMPT-induced dopamine depletion in drug naive schizophrenic patients, indicating a dysregulation of dopaminergic neurotransmission. Results of studies combining amphetamine challenge and the NMDA receptor antagonist ketamine are related to glutaminergic dysfunction and neurotransmitter interactions. FMRI and PET results demonstrating alterations in task-specific functional connectivity between brain areas are discussed with a focus on the prefrontal cortex and temporal structures. Increase of serotonin-1A receptor binding potential in prefrontal and mesotemporal cortex is related to the serotonin-dopamine interaction. Genetic neuroimaging techniques, including voxel-based morphometry (VBM) and fMRI, revealing significant effects of the dopamine metabolizing enzyme COMT on functional activation in prefrontal areas are also discussed. Functional neuroimaging based on challenge-paradigms in PET as well as task-specific state- or trait-dependent alterations of activation patterns in fMRI, seems to be a promising candidate for the development of biological marker tests for schizophrenia. PMID- 16270247 TI - [Change in social networks due to psychoses]. AB - The paper presents the network concept and filters research results according to practical relevance. By way of introduction, the concept of the social network SNW is briefly outlined in terms of its origins and significance. The "socially intact" network of 25 - 40 individuals, of which 15 - 20 % are very close and familiar, is compared with that of people suffering from schizophrenia, averaging 13 individuals. The changes are compiled according to the individuals involved, the type of relationship, and the type of perception. Outcome-relevant results of the network research are summarised on a general basis and according to the parameters social support, social and family contacts, distribution by sex, symptoms, and relatives. The changes in the network in the course of the disease and their importance are described separately according to the phases premorbid phase, prodromal phase, early psychosis, untreated psychosis, first phase of treatment, and long-term phase, illustrating their importance in the context of the overall pathological process. Possibilities of intervention in the SNW are outlined for the phases premorbid, prodromal, onset of the first episode, chronicity development, and existing chronicity. The conclusion rounds off with a brief enumeration of the often lacking intervention options in the German speaking world. PMID- 16270246 TI - [Behavioral concepts for the etiology of schizophrenia]. AB - Schizophrenic disorders are a relatively common and severe psychiatric disorder. Extensive research efforts during the last two decades could show that the course and outcome of the disorder will be determined by a number of biological, psychological and social factors. The vulnerability-stress-coping-competence model as a tentative heuristic framework integrates the different factors which contribute to the understanding of the disorder. In this model psychological factors play an important role. Some of these factors are subject of this paper: expressed emotions and the cognitive understanding and treatment of delusions and hallucinations. PMID- 16270248 TI - [Etiological concepts of chronification in schizophrenia]. AB - Kraepelin and Eugen Bleuler understood the chronicity of schizophrenia as a typical expression of its natural course. Although they erred in this pessimistic assessment, some 30-35 % of all schizophrenia patients do suffer from chronic residual symptoms after years of having the disease, despite sociopsychiatric reform efforts that led to open hospitals and greater patient autonomy. According to the vulnerability conception, schizophrenia is regarded as a vulnerability that causes decompensation leading to psychotic episodes under the influence of internal and/or external stressors. The vulnerability threshold can be increased or lowered by influencing variables. In addition to factors immanent to the disease, some of these variables are related directly or indirectly to chronicity of the disease. PMID- 16270249 TI - [Psychoanalytical aspects regarding etiology and understanding of schizophrenic psychoses]. AB - The author -- specialized in Psychiatry and Neurology as well as a Psychoanalyst - reports her experience during some decades of treating patients with schizophrenia in greatly different hospitals and institutions. Many examples demonstrate that psychoanalytical knowledge and recognizing psychodynamic connections can essentially contribute to understanding and treating psychotic patients. From the multitude of psychoanalytic theories on the etiology of schizophrenia some aspects are demonstrated, mainly those regarding their use in practical work. Some of them are: ego-disturbance, the psychotic symptom as a defence mechanism and the problem of countertransference. Regarding the ego disturbance some examples show that usually not all ego-functions are impaired. Thus, cognitive functions may not only frequently be preserved, but may be working even better than usual. Out of numerous mechanisms of defense in schizophrenia projection, identification, projective identification, splitting, denial and regression are especially important. Moreover, psychotic patients frequently show certain symptoms that are related to very personal experiences. They can often be recognized quite easily as defence mechanisms. Countertransference is of great importance when treating such patients. Three of these numerous phenomenous are mentioned especially: anxiety, compassion, which may lead to identification, and the incapacity to accept the limit of therapeutic measures. The author is convinced that the so-called "psychodynamic" and "biological" psychiatry are not in opposition. Splitting them is not justified because according to the different stages of illness treatment with drugs as well as with different psychotherapeutic methods, even with psychoanalytical ones and social rehabilitation, are necessary. The etiology of schizophrenic psychosis cannot be explained with psychoanalysis either. Yet, the knowledge of psychodynamic processes can contribute essentially to understanding the illness and treating the patient. The compliance accomplished thereby can improve the quality of life of the patient and possibly for the therapist too. PMID- 16270250 TI - [Poetry and schizophrenia]. AB - The confrontation of a poet with himself and his world may be obvious in his creations. When a breakdown like a schizophrenic illness occurs in his life he will fight with creative efforts against his injury. As fragment of this struggle poetic expressions result which allow us to understand the inner life of a psychotic disturbed person better. The poems of two Austrian authors of our time can help us as introduction in this field. PMID- 16270251 TI - [A systemic view on psychosis]. AB - The experience of time changes during life. In the youth time changes slowly from the longitudinal point of view, looking cross on life very fast. In older ages regularly it is the opposite matter, although there are throughout all ages significant intra- and interindividually differences. Experience of time is highly sensitive towards changes of one's self-experience, which can be well observed in psychiatric disorders. In case of psychosis identity of time and experienced synchronism (as certainty to be in the time with others) will be destroyed. There is a kind of breaking up time, a compression of time, where there is no before nor after. In depressed people time experience is similar to elder people, accordingly manic patients feel something like children in this sense. Dissociation of time has got a key role for understanding psychosis from the systemic point of view. Synchrone dissociation is distinguished from diachrone dissociation. In the first case conflicts will be "solved" through excessive simultaneity, undercutting the minimal time interval. The possibilities of differences are made impossible. Secondly the maximum of the time interval is exceeded; therefore the integration of opposite aspects is impossible. Equally no conflict will be possible. Both kinds of dissociation lead to "pseudo-harmony" of interrelational systems, despite of potency for conflicts. In family systems with schizophrenic members synchrone dissociation is typical, whereas diachrone dissociation is common in those with affective disorders. Both of them will be found in communicative systems with patients suffering from schizoaffective disorders. In between occur fierce arguments, when "protection" of dissociation is not granted. PMID- 16270252 TI - [Theories on etiopathogenesis of psychoses in childhood and adolescence]. AB - OBJECTIVE: A focused review of a selected part of research on phenomenology of psychoses in children and adolescents is provided. Especially the differences to psychoses in adults are examined. Hallucinations and delusions are illustrated in context to the stage of development. RESULTS: The symptoms of the childhood psychotic disorder differ qualitatively and quantitatively from the adult form. Interpretation of symptoms has always to be set in relationship to developmental age. Delusions and hallucinations are referred to the stages of development according to Piaget. A possible explanation for the fact that symptoms vary strongly according to developmental age is provided. DISCUSSION: The insight for the development of psychotic symptoms from the background of child developmental stages will be increased. But many questions about the specific clinical picture of childhood psychoses still remain to be answered. PMID- 16270253 TI - [Late adolescence and young adulthood -- their etiopathogenetic significance for schizophrenia]. AB - The significant upsurge of schizophrenic illness manifestations during adolescence and young adulthood is a core finding of epidemiological research in schizophrenia. The reasons are probably manifold, so far, however, not well understood. From the perspective of neurobiological maturation, processes of neuronal reorganisation in cortical and subcortical structures are hot topics. Aberrant forms of pruning, sprouting and myelinization may play a major role in the pathogenesis of a schizophrenic breakdown. The consequences for the pathophysiology of schizophrenia have to be considered. Models of neuroplasticity during adolescence can be connected with models of a neurodevelopmental vulnerability and models of neurotoxicity within an integrated aproach in order to better understand premorbid adjustment, onset, and course of schizophrenic illness. From the perspective of psychosocial development, prominent developmental tasks have to be considered, which form a major challenge to all adolescents, bearing, however, special risks for some adolescents. Patterns of psychosocial adaptation found during this developmental period must be considered in their long-term consequences. Neurobiological reorganisation and psychosocial transformation are both sides of one developmental process during adolescence and young adulthood. PMID- 16270254 TI - [Diagnosis and therapy of paranoid syndromes in the elderly]. AB - Paranoid syndromes in the elderly are nosologically unspecific. Delusional disorder is characterized by the presence of delusions, typically persecutory, commonly in their homes. Frequent risk factors are discriminating, insulting or threatening life events, social isolation as well as sensory impairments. Atypical antipsychotics are considered as first line treatment. Delusion of parasitosis and erotomania are presented as subtypes of delusional disorders. PMID- 16270255 TI - [Risk factors for delinquent behavior in schizophrenic patients -- a survey from Krafft-Ebing till today]. AB - The dangerousness of psychotic, especially schizophrenic patients has always been a matter for controversial discussion. On the one side psychiatrists got under suspicion to show too much understanding for deviant and aggressive acts of their patients and to forget the public need for security. On the other side experts disagreed in their opinion on the risk of aggressive behaviour. The following paper reviews the development in evaluation of risk factors from Krafft-Ebing to the presence. On this way the studies of Boker are a milestone in forensic psychiatry. The doctrine "schizophrenic = irresponsible = dangerous" was not valid any more. He showed that the risk for violence depends on much more than the diagnosis of mental illness. Based on the outcome of his study and his differentiated descriptions of subtypes of schizophrenia and their individual risk of violence modern psychological prognostic instruments were developed. PMID- 16270256 TI - The Coombs' test in veterinary medicine: past, present, future. AB - The Coombs' test, also known as the antiglobulin test, is used most frequently in veterinary medicine as an aid in the diagnosis of immune-mediated hemolytic anemia. The test also is used widely in human medicine to screen for red blood cell alloantibodies. Polyspecific reagents for veterinary use typically contain anti-IgG, anti-IgM, and anti-C3. Monospecific reagents also are available. False positive and false-negative test results can be obtained. Inadequate sensitivity of the standard test in human and veterinary medicine has necessitated development of alternate, more sensitive technologies. PMID- 16270257 TI - An immunoturbidimetric assay for rapid quantitative measurement of feline alpha-1 acid glycoprotein in serum and peritoneal fluid. AB - BACKGROUND: Alpha-1-acid glycoprotein (AGP) is an acute phase protein that increases in concentration in infectious and inflammatory conditions. The serum and peritoneal fluid concentrations of AGP may be useful in the diagnosis of feline infectious peritonitis (FIP), a lethal disease of cats. Currently AGP can be measured by radioimmunodiffusion (RID) assays, which are time consuming and difficult. OBJECTIVES: The objectives of this study were to develop a rapid immunoturbidimetric assay for measurement of AGP in feline serum and peritoneal fluid and to compare the results with those obtained by RID. METHODS: AGP was purified by perchloric acid precipitation and ion-exchange chromatography from a pool of peritoneal fluid obtained from cats with FIP, as determined by a panel of laboratory tests, including serum AGP concentration, albumin: globulin ratio, and total protein concentration, anti-coronavirus antibody titers, and effusion analysis. The purified AGP in a complete Freund's adjuvant and Tween 20 mixture was injected into a sheep and blood was collected at monthly intervals. Anti-AGP antiserum, as confirmed by ELISA and Western blot techniques, and a pool of peritoneal fluid from cats with FIP were used to prepare standards. Clinical samples of feline peritoneal fluid (n=55) and serum (n=59) were assayed for AGP and results from the immunoturbidimetric and RID methods were compared. RESULTS: Significant correlation (P < .001) was obtained between methods for both peritoneal fluid (R2=.9259) and serum (R2=.9448) samples. Coefficients of variation for the immunoturbidimetric method were <5%. CONCLUSIONS: This rapid immunoturbidimetric assay for measurement of feline AGP in serum and peritoneal fluid may be of value in the diagnosis of FIP and possibly other inflammatory diseases in cats. PMID- 16270258 TI - Multiple myeloma in 16 cats: a retrospective study. AB - BACKGROUND: There is limited published information regarding feline multiple myeloma. Diagnostic criteria are derived from canine studies and to our knowledge, have not been critically reviewed for cats. OBJECTIVE: To evaluate the clinical and laboratory findings in cats with multiple myeloma and appraise diagnostic criteria. METHODS: Retrospective evaluation of medical records was performed. Inclusion required an antemortem diagnosis of multiple myeloma using 2 of 4 criteria: 1) >or=20% plasma cells in the bone marrow, or >or=10% if atypical plasma cells; 2) paraproteinemia; 3) radiographically-evident osteolysis; 4) light chain proteinuria. Alternatively, a postmortem diagnosis was based on the findings of multiple plasma cell neoplasms, with marrow involvement. RESULTS: Sixteen cats were diagnosed with multiple myeloma between 1996 and 2004, with a median age of 14.0 years; 9 of 16 (56%) were castrated males, and 7 of 16 (44%) were spayed females. Laboratory abnormalities included hyperglobulinemia (14/16, 87.5%), with 11/14 (78.5%) monoclonal and 3/14 (21.4%) biclonal gammopathies; hypoalbuminemia (4/16, 25%); light chain proteinuria, (4/9, 44.4%); hypocholesterolemia (11/16, 68.7%); hypercalcemia, (3/15, 20%); nonregenerative anemia, (11/16, 68.7%); regenerative anemia, (1/16, 6.2%); neutropenia (5/15, 33.3%); thrombocytopenia (8/16, 50%); and marrow plasmacytosis (14/15, 93.3%). Plasma cells were markedly immature, atypical, or both in 10 of 12 (83.3%) cats. Focal or multifocal osteolysis was noted in 6 of 12 (50%) cats for which radiographs were available for review; generalized osteopenia was found in 1 (8.3%) cat. Noncutaneous, extramedullary tumors were found in all cats assessed, 7/7 (100%), including spleen (6), liver (3), and lymph nodes (4). The disease in 1 of 2 cats with cutaneous tumors progressed to plasmacytic leukemia. CONCLUSIONS: Common findings in feline multiple myeloma include atypical plasma cell morphology, hypocholesterolemia, anemia, bone lesions, and multi-organ involvement. Based on the results of this study, we advocate modifying diagnostic criteria in cats to include consideration of plasma cell morphology and visceral organ infiltration. PMID- 16270260 TI - Type of smear may influence thrombopoietic cell counts in the bone marrow of clinically healthy dogs. AB - BACKGROUND: The expected number of thrombopoietic cells in normal canine bone marrow is poorly defined and there is no consensus on the most appropriate way to prepare cytologic smears to evaluate these cells nor on the optimum method for their quantification. OBJECTIVE: The purpose of this study was to determine total and differential counts of thrombopoietic cells in the bone marrow of clinically healthy Beagle dogs by comparing 4 different smear types and bone marrow core biopsies. METHODS: Twenty-two clinically healthy, male Beagle dogs, 10 to 12 months old, were used in the study. Following bone marrow aspiration and core biopsy from the iliac crest, Giemsa-stained smears were prepared by 4 techniques: drop-squash, particle-squash, buffy coat, and fat-layer smears. Thrombopoietic cells were counted in up to 100 low-power fields (LPF, X10 objective) in the aspiration smears and in all possible high-power fields (HPF, X40 objective) in H&E-stained biopsy sections. RESULTS: Mean total thrombopoietic cell counts were 2.76 cells/LPF (drop-squash), 1.55 cells/LPF (particle-squash), 8.05 cells/LPF (buffy coat), and 3.08 cells/LPF (fat-layer). Core biopsies yielded 5.31 cells/HPF but frequently failed to provide interpretable specimens. There was a significant difference in cell counts among the 4 smear types (P <.001). Based on evaluation of buffy coat smears, thrombopoietic cells included 1.23% megakaryoblasts, 8.77% promegakaryocytes, and 90% megakaryocytes, with a mean maturation index of 0.11. CONCLUSION: Thrombopoietic cell counts in canine bone marrow are influenced by the smear technique. Buffy coat and fat-layer smears may be useful to obtain cellular smears in hemodiluted or small aspirate samples. PMID- 16270259 TI - Oxidative stress in the erythrocytes of cattle intoxicated with Senecio sp. AB - BACKGROUND: Intoxication caused by Senecio sp is characterized by irreversible damage to liver cells and may be associated with oxidative stress. OBJECTIVES: The aim of this study was to evaluate the effects of intoxication by Senecio sp on lipoperoxidation, antioxidant defenses, and the osmotic resistance of erythrocytes in cattle. METHODS: Blood samples from 30 intoxicated animals (group 1) and 30 healthy animals (group 2) were analyzed. The diagnosis of poisoning by Senecio sp was based on histopathologic lesions verified through hepatic biopsy. The following biochemical parameters of oxidative stress in the erythrocytes were determined: thiobarbituric acid-reactive substances (TBARS), copper-zinc superoxide dismutase (CuZnSOD) activity, and nonprotein sulfhydryl (NPSH) groups. Erythrocyte osmotic fragility also was evaluated. RESULTS: TBARS concentration and CuZnSOD activity were significantly (P <.001) higher in group 1 when compared with group 2. The concentration of erythrocyte NPSH groups was significantly (P <.03) lower in group 1 when compared with group 2. Osmotic fragility was more pronounced in the erythrocytes of group 1 when compared with group 2 (P < .001). CONCLUSIONS: The results of this study indicate that poisoning by Senecio sp causes an increase in lipoperoxidation, oxidation of NPSH groups, and consequently, oxidative stress in bovine erythrocytes that may contribute to hemolysis. These findings may contribute to a better understanding of the mechanisms involved in cell damage in animals intoxicated by Senecio sp. PMID- 16270261 TI - Long-term fluctuations and effect of age on serum concentrations of certain fat soluble vitamins in dairy cows. AB - BACKGROUND: There is insufficient evidence in the literature concerning the fluctuations and correlations of serum fat-soluble vitamin concentrations during lactation in dairy cows and the potential effect of age of the animals on these values. OBJECTIVES: The objectives of this study were to determine fluctuations in mean serum concentrations of the fat-soluble vitamins A and E and b-carotene and the correlations among them in clinically healthy dairy cows, from the last month of the dry period until the end of lactation, and to investigate whether age was a factor that affects these serum concentrations. METHODS: Forty-five clinically healthy Holstein cows were assigned to 2 groups based on age. Group A consisted of 23 cows 4 years old (mean +/- SD, 3.24 +/- 0.44 years), and group B consisted of 22 cows >4 years old (6.68 +/- 1.66 years). Blood samples for determination of serum beta-carotene, vitamin A, and vitamin E concentrations were collected from each animal just before the start of study (1 month prior to expected parturition), at parturition, and thereafter, at monthly intervals until the end of lactation. RESULTS: There was a significant positive correlation between beta-carotene, vitamin A, and vitamin E values. Mean serum concentrations of beta-carotene, vitamin A, and vitamin E were significantly higher in younger animals. CONCLUSIONS: To our knowledge, this is the first evidence that age is a factor affecting serum concentrations of fat-soluble vitamins in dairy cows. PMID- 16270262 TI - Sensitivity of Tru-cut and fine needle aspiration biopsies of liver and kidney for diagnosis of feline infectious peritonitis. AB - BACKGROUND: The detection of typical lesions and feline coronavirus (FCoV) antigen in tissues is the only conclusive method for making a diagnosis of feline infectious peritonitis (FIP). A positive result using Tru-cut biopsy (TCB) and fine-needle aspiration biopsy (FNAB) has high diagnostic specificity, but information about the capacity of these techniques to correctly identify cats with FIP lesions is not available. OBJECTIVES: The diagnostic sensitivity of TCB and FNAB for detecting liver and kidney histologic lesions caused by FIP was evaluated. METHODS: TCB and FNAB specimens collected mainly at necropsy from 25 cats with FIP were analyzed. Diagnostic sensitivity was calculated on the basis of the number of false-negative and true-positive specimens, compared with the number of organs bearing histologic lesions of FIP. RESULTS: Diagnostic sensitivity was higher for hepatic TCB (64%) and FNAB (82%) than for renal (39% and 42%, respectively) procedures. A high percentage of renal cytologic and TCB specimens were inadequate. Combined analysis of TCB and FNAB specimens collected from the same organ increased the diagnostic sensitivity for liver (86%) and kidney (48%). The sensitivity of immunohistochemical/cytochemical analysis was low (11-38% depending on the technique), probably due to variable distribution of feline coronavirus in the lesions. CONCLUSION: Biopsy of liver and kidney can correctly identify FIP lesions. However, false-negative results or inadequate samples occur with moderate frequency, especially for immunochemical analysis. Diagnostic sensitivity may be increased when both TCB and FNAB specimens from the same organ are examined. PMID- 16270264 TI - Temporal effects of 3 commonly used anticoagulants on hematologic and biochemical variables in blood samples from macaws and Burmese pythons. AB - BACKGROUND: Few studies have been done to evaluate anticoagulants for use with blood samples from birds and reptiles. Heparin currently is the most commonly used anticoagulant in practice, but may adversely affect blood cell staining and quantitation. OBJECTIVE: The purpose of this study was to evaluate the effects of lithium heparin, K3-EDTA, and sodium citrate, with and without the addition of albumin, on hematologic variables in macaw (Ara sp) and python (Python molurus bivittatus) blood samples. METHODS: Blood samples from 10 macaws and 10 Burmese pythons were collected in heparin-coated syringes and placed into tubes containing either lithium heparin, K3-EDTA, or sodium citrate with and without the addition of 0.25 mL of a 22% bovine serum albumin solution. Cell lysis was determined by counting the number of lysed cells/200 WBCs in Wright's-Giemsa stained blood smears and by qualitative evaluation of pink plasma in microhematocrit tubes. A CBC was done after 3, 12, and 24 hours of storage at 4 degrees C in anticoagulant-containing tubes and results were compared with those obtained at 0 hour for the heparin-coated syringe sample. A biochemical panel also was done at each time point in similarly stored lithium-heparin samples. RESULTS: Hemolysis was significantly increased in citrated samples from both macaws and pythons beginning at 12 hours. At 24 hours, 19 of 30 (63%) macaw samples in all anticoagulants had >100 lysed cells/200 WBCs. There were no significant differences in hematologic values in samples from pythons collected in heparin or EDTA at any time point. No significant differences were found in the number of lysed cells or in other hematologic data in samples with albumin. Glucose concentration decreased and potassium concentration increased significantly over time in heparinized blood samples. CONCLUSIONS: Based on the results of this study, whole blood samples anticoagulated with lithium heparin or EDTA should be evaluated within 12 hours (macaws) or 24 hours (pythons) of collection and stored at 4 degrees C for best results. Citrate should be avoided as it may result in increased cell lysis. The addition of albumin does not prevent cell lysis. PMID- 16270263 TI - Evaluation of latex agglutination kits for detection of fibrin(ogen) degradation products and D-dimer in healthy horses and horses with severe colic. AB - BACKGROUND: Fibrin(ogen) degradation products (FDPs) and D-dimer are sensitive indicators of excessive fibrinolysis due to disseminated intravascular coagulation (DIC) in dogs. To the authors' knowledge, latex-agglutination-based plasma FDP and D-dimer assays have not been validated for use in horses. OBJECTIVES: To determine: 1) sensitivity and specificity of latex agglutination serum and plasma FDP and D-dimer assays for diagnosis of DIC; and 2) their prognostic value in horses with severe colic. METHODS: At hospital admission and 24 hours later, blood was collected from 30 healthy horses and 20 horses with severe colic. Horses fulfilling predefined laboratory criteria of DIC were enrolled, and their data were subcategorized by survival for analysis. Platelet counts were determined and coagulation panel testing was performed. Serum and plasma FDP concentrations were measured using separate latex agglutination kits. Plasma D-dimer concentration was measured using 3 latex agglutination kits and a card immunofiltration test. Test sensitivity and specificity results were determined for healthy horses and those with colic. Median test values were compared between colic survivors and nonsurvivors to evaluate the prognostic usefulness of all tests. RESULTS: Performance characteristics varied among assays and kit suppliers. The FDP assays had low sensitivity (<40%), whereas the most accurate D-dimer kit had 50% sensitivity and 97% specificity. High D-dimer concentration was the third most common hemostatic abnormality in horses with colic. Median antithrombin (AT) activity was significantly lower and activated partial thromboplastin time (aPTT) was significantly longer in nonsurvivors than survivors. CONCLUSIONS: Commercial latex-agglutination D-dimer assays might prove useful as adjunctive tests for the diagnosis of DIC in horses with severe colic; however FDP assays are invalid for this purpose. Low AT activity and prolonged aPTT at admission are associated with a poor prognosis in this patient population. PMID- 16270265 TI - Validation of human recombinant tissue factor-activated thromboelastography on citrated whole blood from clinically healthy dogs. AB - BACKGROUND: Thromboelastography (TEG) is an analytical method that enables global assessment of hemostatic function in whole blood (WB) with evaluation of both plasma and cellular components of hemostasis. TEG has a largely unused potential in the diagnostic workup and monitoring of dogs with hemostatic disorders and it may be a valuable supplement to traditional coagulation parameters. OBJECTIVES: The objective of this study was to establish a clinically applicable reference interval for a TEG assay using recombinant human tissue factor (TF) as the activator on citrated WB from clinically healthy dogs and to evaluate the stability of citrated WB stored for 30 minutes (T30) and 120 minutes (T120) at room temperature (RT). Additionally, we evaluated the analytical variation in reaction time (R), clotting time (K), angle (alpha), and maximum amplitude (MA). METHODS: Blood was collected from 18 clinically healthy dogs. Duplicate TEG analyses with TF as the activator at a concentration of 1:50,000 were performed on canine citrated WB at T30 and T120. R, K, a, and MAwere analyzed. RESULTS: Mean TEG values at T30/T120 were R = 5.61/4.91 minutes, K = 4.20/3.34 minutes, alpha = 45.33/50.90 degrees , and MA = 47.96/50.19 mm. Significant differences in these values were observed after storage for T30 and T120 at RT, with a tendency towards hypercoagulability at T120. The mean coefficients of variation were low. CONCLUSIONS: Canine citrated WB can be used for TEG analysis with human recombinant TF as the activator when stored at RT for T30 or T120. At both time points, the analytical variation was low, suggesting that TEG analysis may be of value in evaluating dogs with hemostatic disorders. A fixed time point should be chosen for serial measurements. PMID- 16270266 TI - Characterization and quantification of blood cells from the umbilical cord of dogs. AB - BACKGROUND: Models for the study of hematopoietic stem cells in dogs provide important information for bone marrow transplantation in humans. Recent studies have reported the importance of human umbilical cord blood (UCB) as an alternative to allogenic bone marrow for hematopoietic reconstitution. However, there are no studies on the UCB cells of dogs. OBJECTIVE: The aim of this experiment was to characterize and quantify the blood cells of the umbilical cord of dogs. METHODS: The blood of the umbilical cord of 20 neonatal dogs, delivered at term, with a median gestation time of 58 days, was collected with a 5-mL syringe containing EDTA. Total RBC, WBC, and platelet counts, HCT, hemoglobin (Hgb) concentration, and RBC indices were determined using an automatic cell counter. The differential leukocyte count was determined manually in blood smears stained with May-Grunwald-Giemsa. Reticulocyte percentages were determined on blood smears stained with brilliant cresyl blue and counterstained with May Grunwald Giemsa. RESULTS: The MCHC and numbers of RBCs, WBCs, neutrophils, and eosinophils in UCB were lower as compared with reference values for the peripheral blood of healthy neonatal and adult dogs; whereas, the MCV and reticulocyte percentages were higher. CONCLUSION: Erythrocyte macrocytosis and hypochromasia in UCB were consistent with marked reticulocytosis and indicative of high erythropoietic activity. The results of this study are an important first step in the characterization of UCB from neonatal dogs. PMID- 16270267 TI - Serum protein electrophoresis in retired racing Greyhounds. AB - BACKGROUND: Retired racing Greyhounds are becoming common as pets. Because of their unique physiology, results of routine laboratory tests are frequently outside the reference interval for dogs. Compared with other breeds, Greyhounds have low serum protein concentrations, but the concentrations of different serum protein fractions have not been reported. OBJECTIVES: Our objectives were to evaluate the results of serum protein electrophoresis (SPE) in healthy, retired racing Greyhounds and compare them with a control group of age- and gender matched non-Greyhound dogs. METHODS: Agarose gel electrophoresis was done using a standard method; the gels were stained with amido black and scanned with a Cliniscan 2 densitometer (Helena Laboratories, Beaumont, TX, USA). Protein fractions were identified by visual inspection of the electrophoretogram. A Student's t-test assuming equal variances was used to compare the concentration of the different fractions between groups. RESULTS: The concentrations of total protein, total globulins, and alpha-1-, alpha-2-, beta-1-, and beta-2-globulins were significantly lower and the albumin to globulin (A:G) ratio was significantly higher in Greyhounds than in non-Greyhound dogs (P < .05). There was no significant difference in albumin or gamma-globulin concentrations. CONCLUSIONS: Low serum protein concentrations in Greyhounds are the result of low concentrations of a- and b-globulins. These results should be kept in mind when evaluating both healthy and sick Greyhounds. Additional studies are needed to identify the individual proteins associated with low alpha- and beta-globulin concentrations in Greyhounds. PMID- 16270268 TI - Bronchogenic adenocarcinoma in a cat: an unusual case of metastasis to the skin. AB - A 6-year-old, spayed, female, domestic shorthair cat was presented for decreased activity. A nodular lesion was found in the skin extending into the subcutaneous tissue of the right abdominal flank. On lateral and ventrodorsal radiographs of the thorax, an opacity involving the entire right caudal lung lobe and pleural effusion were noted. Cytologic evaluation of cells in the thoracic fluid and in the mass revealed a population of atypical epipthelial cells with marked anisocytosis and high N:C ratios, organized in acinar-like clusters. Multinucleated cells and several mitotic figures were found. The cytologic interpretation was carcinoma. Because of the progressive severity of clinical signs, the cat was euthanized. Histologic evaluation of tissues obtained at necropsy indicated a bronchogenic adenocarcinoma in the lung, with metastasis to the skin of the right flank, but no involvement of the digits. Based on immunohistochemical stains, the neoplastic cells strongly co-expressed cytokeratin and vimentin, and were negative for S-100 and actin-specific antigen. Bronchogenic adenocarcinoma is an uncommon neoplasm in cats, and the digits are the most common sites of metastasis. This case was unusual in that the skin of the abdominal wall was the primary site of metastasis, with no involvement of the digits. PMID- 16270269 TI - Thyroid follicular adenocarcinoma in a ferret. AB - A 5-year-old male castrated ferret was presented to the Washington State University College of Veterinary Medicine for evaluation of progressive hair loss and a large, rapidly growing ventral neck mass. The patient had been diagnosed previously with an insulinoma, which was managed medically. Fine-needle aspirates of the neck mass were performed. The cytologic results were most consistent with epithelial neoplasia, likely a carcinoma; thyroid origin was considered likely based on tumor location and cell morphology. The tumor grew rapidly, and the owners elected euthanasia 1 week after examination. At necropsy, a circumscribed, ovoid mass disrupted the right cervical musculature next to the right lobe of the thyroid gland. Histopathologic evaluation revealed an infiltrative mass consisting of cuboidal cells arranged in solid sheets and irregular follicles enclosing colloid. The cells were large, with prominent nucleoli, and had a high mitotic rate. The histopathologic diagnosis was consistent with thyroid follicular adenocarcinoma. Immunochemical findings confirmed thyroglobulin production by neoplastic cells, but to a lesser extent than in normal ferret thyroid tissue. To our knowledge, this is the first case of thyroid follicular adenocarcinoma to be reported in a ferret, with only 1 other case of thyroid carcinoma, a C-cell carcinoma, described previously. PMID- 16270270 TI - Diagnosis of systemic cryptococcosis by fecal cytology in a dog. AB - A 3-year-old Boxer was presented with progressive diarrhea, vomiting, and lethargy of 5-months duration. The dog had watery black feces, a mature neutrophilia, and microcytic anemia. Cytologic evaluation of a direct fecal smear stained with Wright's-Giemsa revealed numerous encapsulated, narrow-based, budding organisms consistent with Cryptococcus sp. Pyogranulomatous inflammation and Cryptococcus organisms also were observed in ultrasound-guided fine-needle aspirates of the small intestine and mesenteric lymph nodes, and in histologic sections of colonic biopsies obtained by endoscopy. Multifocal chorioretinitis by fundic examination was consistent with systemic mycosis, and the reciprocal antigen titer (1600) on a cryptococcal antigen latex agglutination test for Cryptococcus neoformans was markedly increased. Using immunohistochemistry, the organism was identified further as C neoformans var. grubii (C neoformans var. neoformans serotype A). After 3 weeks of antifungal treatment, ultrasound examination revealed urinary bladder wall thickening, and Cryptococcus organisms were found in a urine sediment preparation. After 4 months of treatment, the dog was clinically normal and had no abnormal findings on CBC, serum biochemistry, urinalysis, or fecal cytology; however, the antigen titer remained unchanged, mesenteric lymphadenomegaly and jejunal wall thickening were still evident, and cytologic evaluation of fine-needles aspirates of the jejunal wall revealed budding Cryptococcus organisms. Intestinal involvement in dogs with cryptococcosis is rare, and diagnosis by fecal cytology has not been documented previously. PMID- 16270271 TI - Acremonium strictum pulmonary infection in a horse. AB - A 10-year-old Thoroughbred gelding was admitted to the Veterinary Medical Teaching Hospital of the University of California-Davis with a 2-week history of intermittent fever and acute onset of lethargy, anorexia, and ataxia. Although the clinical signs were nonspecific, the results of initial hematologic and biochemical analysis were consistent with a chronic inflammatory process. Thoracic radiographs revealed an increased fine reticulonodular interstitial opacity throughout the dorsal caudal lung fields. Cytologic examination of bronchoalveolar lavage (BAL) fluid showed mixed inflammation with many mononuclear phagocytes containing single, spherical, intracytoplasmic fungal organisms. Four mold species were cultured in low numbers from the BAL fluid. One of the fungal elements observed on the culture plates was identified as Acremonium strictum by real-time polymerase chain reaction (PCR). A diagnosis of fungal pneumonia due to A strictum was made based on the results of thoracic imaging, cytologic evaluation, culture, and PCR testing. The horse made an uneventful recovery with supportive treatment and was disease-free based on normal physical, radiographic, and cytologic findings at 21 days after presentation. To our knowledge, this is the first report of isolation of A strictum from the BAL fluid of a horse with interstitial pneumonia. PMID- 16270272 TI - Of all the nerve! A subcutaneous forelimb mass on a cat. AB - A 16-year-old, male, neutered cat had a 2.5 X 1.5 cm mass on the medial aspect of the right carpus. Cytologic examination of a fine-needle aspirate of the mass indicated a markedly pleomorphic population of plasmacytoid to histiocytic appearing cells. The cytologic diagnosis was malignant neoplasia of probable mesenchymal or round cell origin. The right forelimb was surgically removed and the scapular, axillary, and prescapular lymph nodes were excised. Malignant fibrous histiocytoma was tentatively diagnosed histologically; however, the tumor cells subsequently were found to be negative for histiocytic (MAC 387, antitrypsin), T-cell (CD3), and B-lymphocyte (immunoglobulin light chains, Ly 5/CD45R) markers, and positive for glial fibrillary acidic protein, vimentin, and S-100. Based on the immunohistochemical results, the diagnosis was modified to malignant peripheral nerve sheath tumor (PNST). Six months after surgery, the cat was reported to be well and had no evidence of metastasis. PNSTs are rare tumors in cats, and are considered as synonymous with schwannomas, neurofibrosarcomas, and hemangiopericytomas. In this cat, the plasmacytoid and pleomorphic appearance of the PNSTcells in cytologic and histologic specimens was unusual, and made it difficult to reach an accurate diagnosis without immunocytochemistry. PMID- 16270273 TI - Cutaneous mass aspirate from a Golden Retriever: "glandular guile". AB - A 3-year-old, neutered, male Golden Retriever was presented for evaluation of a 10 X 9 X 5 mm, firm, red, raised, cutaneous mass located over the left cranial thorax and noted incidentally by the owner. On cytologic evaluation of a fine needle aspirate of the mass, the interpretation was a malignant tumor with predominantly mesenchymal features. Differentials included liposarcoma, atypical amelanotic melanoma, anaplastic sarcoma, and anaplastic carcinoma. Following complete excision of the mass, a diagnosis of sebaceous adenocarcinoma was made based on histologic features, positive immunostaining for pancytokeratin, and negative staining for vimentin, Melan-A, and S-100. There was no evidence of metastasis on physical examination or thoracic radiographs, and the prognosis was good. The unique and previously unreported cytologic features of this small, sebaceous adenocarcinoma were the extreme pleomorphism, including marked anisocytosis, anisokaryosis, and multinuclearity, and the paucity of epithelial features. PMID- 16270274 TI - Urine sediment from a Chihuahua. AB - A 6-year-old, intact male Chihuahua was presented with stranguria and painful urination of 5 days duration. Cystine crystals were observed in low numbers in unstained urine sediment preparations, and a diagnosis of cystinuria was made. Uroliths were removed surgically from the urethra and the bladder, and mineral analysis indicated the stones were composed of 100% cystine. Cystinuria results from an inherited defect in renal tubular transport of cystine that affects many breeds and has been found as an autosomal recessive trait in Newfoundlands. Accurate identification of cystine crystals in urine is an important means of diagnosing cystinuria. PMID- 16270275 TI - Multilobulated "flower" cells in a subcutaneous mass aspirate from a cat. AB - An 8-year-old intact male cat was presented with a subcutaneous mass in the region of the right jugular vein. Cytologic and histopathologic examinations revealed cells with multilobulated nuclei (flower cells). Immunochemistry using a panel of markers showed vimentin-positivity on cytologic specimens, and postive staining for CD79a and BLA36 on histologic specimens. The final diagnosis was lymphoma of B-cell origin. We have observed similar multilobulated cells in ascites fluid, thoracic fluid, and peripheral blood from dogs and cats with a variety of lymphoid and myeloid neoplasms. Cells with multilobulated nuclei that resemble flower petals also have been described in humans. These cells are infrequently observed in canine and feline cytology specimens and require immunochemistry to determine their cell of origin. PMID- 16270276 TI - The role of lipoxygenase-isoforms in atherogenesis. AB - Lipoxygenases (LOXs) form a heterogeneous family of lipid-peroxidizing enzymes, and several LOX-isoforms (12/15-LOX, 5-LOX) have been implicated in atherogenesis. However, the precise role of these enzymes is still a matter of discussion. 12/15-LOXs are capable of oxidizing lipoproteins (low-density lipoprotein (LDL), high-density lipoprotein (HDL)) to atherogenic forms, and functional inactivation of this enzyme in murine atherosclerosis models slows down lesion formation. In contrast, rabbits that overexpress this enzyme were protected from lesion formation when fed a lipid-rich diet. To contribute to this discussion, we recently investigated the impact of 12/15-LOX overexpression on in vitro foam cell formation. When 12/15-LOX-transfected J774 cells were incubated in culture with modified LDL, we found that intracellular lipid deposition was reduced in the transfected cells when compared with the corresponding control transfectants. This paper briefly summarizes the current status of knowledge on the biological activity of different LOX-isoforms in atherogenesis and will also provide novel experimental data characterizing the role of 12/15-LOX in cellular LDL modification and for in vitro foam cell formation. PMID- 16270277 TI - 4-Hydroxynonenal and cholesterol oxidation products in atherosclerosis. AB - 4-Hydroxynonenal (HNE) is by far the most investigated aldehydic end-product of oxidative breakdown of membrane n-6 polyunsaturated fatty acids. Its potential involvement in the pathogenesis of atherosclerosis has been corroborated by its consistent detection in both oxidized LDL and fibrotic plaque in humans. HNE has been shown to activate both macrophage and smooth muscle cells, i.e. the two key cell types in chronic inflammatory processes characterized by excessive fibrogenesis. By signalling to the nucleus, the aldehyde may up-regulate in these cells both expression and synthesis of monocyte chemotactic protein 1 (MCP-1) and transforming growth factor beta1 (TGFbeta1). Oxysterols, namely 27 carbon atoms oxidation products of cholesterol, are found in relatively high amount in LDL from hypercholesterolemic individuals and are consistently detectable in foam cells and necrotic core of human atherosclerotic lesion. As for HNE, the challenge of cells of the macrophage lineage with a mixture of oxysterols like that detectable in hypercholesterolemic individuals led to a marked overexpression of TGFbeta1 and MCP-1. Both HNE and oxysterols then appear to be candidates for a primary role in the progression of the atherosclerotic process. PMID- 16270278 TI - Oxidized phospholipids, isolevuglandins, and atherosclerosis. AB - Autoxidation of polyunsaturated phosphatidylcholines (PCs) generates isolevuglandins (isoLGs) through rearrangements of isoprostanoid endoperoxides. Within seconds, isoLGs are sequestered by covalent adduction with proteins. Murine plasma isoLG-protein levels increased at least 2.5-fold in response to inflammation. IsoLG-protein adducts accumulate in vivo providing a convenient dosimeter of oxidative stress. Elevated blood isoLG-protein levels present in atherosclerosis (AS) patients point to an independent defect that is not associated with total cholesterol levels, which results in an abnormally high level of oxidative injury in AS. Protein adduction and cross-linking caused by isoLGs can obstruct protein function. For example, it interferes with proteosomal degradation of proteins and, consequently, may result in apoptotic death of smooth muscle cells and destabilization of atherosclerotic plaques. Phospholipid autoxidation also generates biologically active oxidatively truncated PCs through fragmentation of dihydroperoxydienes that can be promoted by alpha-tocopherol. The oxidatively truncated PCs in oxidized low-density lipoprotein (oxLDL) contribute to the etiology of AS by inhibiting enzymatic activities required for normal processing of oxLDL by macrophages. They promote interactions of monocytes with endothelial cells that may foster migration of monocytes into the subendothelial space. They are also ligands for unregulated receptor-mediated uptake of oxLDL by monocyte macrophages leading to foam cell formation. PMID- 16270279 TI - Oxidized phospholipids as triggers of inflammation in atherosclerosis. AB - Chronic inflammatory diseases including atherosclerosis are major causes of morbidity and mortality worldwide. However, the factors, which trigger processes that determine the outcome of an inflammatory response, are still poorly understood. Accumulating evidence suggests that certain lipid oxidation products, such as oxidized phospholipids (OxPL), may represent endogenously formed factors that are capable of triggering vascular inflammation. This review will address important questions regarding mechanisms involved in acute and chronic inflammation, and discuss the role of OxPL as key players in triggering the inflammatory response in atherosclerosis. Better understanding of how OxPL contribute to vascular inflammation should lead to new strategies in the treatment of chronic inflammatory disorders. PMID- 16270280 TI - The role of dietary oxidized cholesterol and oxidized fatty acids in the development of atherosclerosis. AB - The etiology of atherosclerosis is complex and multifactorial but there is extensive evidence indicating that oxidized lipoproteins may play a key role. At present, the site and mechanism by which lipoproteins are oxidized are not resolved, and it is not clear if oxidized lipoproteins form locally in the artery wall and/or are sequestered in atherosclerotic lesions following the uptake of circulating oxidized lipoproteins. We have been focusing our studies on demonstrating that such potentially atherogenic oxidized lipoproteins in the circulation are at least partially derived from oxidized lipids in the diet. Thus, the purpose of our work has been to determine in humans whether oxidized dietary oxidized fats such as oxidized fatty acids and oxidized cholesterol are absorbed and contribute to the pool of oxidized lipids in circulating lipoproteins. When a meal containing oxidized linoleic acid was fed to normal subjects, oxidized fatty acids were found only in the postprandial chylomicron/chylomicron remnants (CM/RM) which were cleared from circulation within 8 h. No oxidized fatty acids were detected in low density lipoprotein (LDL) or high density lipoprotein (HDL) fractions at any time. However, when alpha-epoxy cholesterol was fed to human subjects, alpha-epoxy cholesterol in serum was found in CM/RM and also in endogenous very low density lipoprotein, LDL, and HDL and remained in the circulation for 72 h. In vitro incubation of the CM/RM fraction containing alpha-epoxy cholesterol with human LDL and HDL that did not contain alpha-epoxy cholesterol resulted in a rapid transfer of oxidized cholesterol from CM/RM to both LDL and HDL. We have suggested that cholesteryl ester transfer protein is mediating the transfer. Thus, alpha-epoxy cholesterol in the diet is incorporated into CM/RM fraction and then transferred to LDL and HDL contributing to lipoprotein oxidation. We hypothesize that diet-derived oxidized fatty acids in chylomicron remnants and oxidized cholesterol in remnants and LDL accelerate atherosclerosis by increasing oxidized lipid levels in circulating LDL and chylomicron remnants. This hypothesis is supported by our feeding experiments in animals. When rabbits were fed oxidized fatty acids or oxidized cholesterol, the fatty streak lesions in the aorta were increased by 100%. Moreover, dietary oxidized cholesterol significantly increased aortic lesions in apo-E and LDL receptor-deficient mice. A typical Western diet is rich in oxidized fats and therefore could contribute to the increased arterial atherosclerosis in our population. PMID- 16270281 TI - Is there a future for antioxidants in atherogenesis? AB - Antioxidants, preferentially those of dietary origin, have for a long time been considered to help against diseases that are presumably aggravated by oxidative stress, such as cardiovascular diseases, cancer, and neurodegenerative disorders. The outcome of clinical trials undertaken to corroborate this hypothesis, however, remained largely inconclusive. Evidence is now emerging that some dietary "antioxidants" influence signaling pathways and the expression of genes relevant in atherosclerosis by mechanisms other than antioxidative ones. By concrete examples we show that (1) vitamin E has gene regulatory functions which might be more important than acting as an antioxidant in vivo, (2) selenium itself is not an antioxidant at all, and even not in general when incorporated into glutathione peroxidases, and (3) a moderate oxidative stress is beneficial rather than detrimental since it can induce defense mechanisms counteracting xenobiotic and oxidative stress. Thus, there is only a future for antioxidants in the prevention of any disease if their real mechanism of action is considered and suitable read-outs and biomarkers are established. PMID- 16270285 TI - Hypercholesterolemia and inflammation in atherogenesis: two sides of the same coin. AB - An abundance of experimental, clinical, and epidemiologic data capped by stunning interventional results with the statins has established hypercholesterolemia as a major causative factor in atherogenesis. In familial hypercholesterolemia and in animal models it is a sufficient cause. Some degree of hypercholesterolemia, perhaps 30-50 mg/dL, may even be a necessary cause. It is equally clear that from the very beginning atherogenesis has a strong inflammatory component, i. e., it is characterized by penetration of monocytes and of T-cells into the developing lesion. These cells, through the secretion of cytokines and growth factors, through immune responses, and through complex cross-talk with elements of the artery wall modulate the growth of the lesion and affect its stability. But inflammation has to occur in response to something. What is that something? What is the "injury" in "response-to-injury"? The case will be made that oxidized lipids in oxidized LDL or generated in response to prooxidative changes in the cells of the artery wall should be considered a plausible candidate. There is no need to consider hypercholesterolemia and inflammation as alternative hypotheses. Both are very much involved. Optimal intervention and prevention will probably require attention to both. PMID- 16270286 TI - The relation of lipid peroxidation processes with atherogenesis: a new theory on atherogenesis. AB - The extremely high sensitivity of polyunsaturated fatty acids (PUFAs) to oxygen is apparently used by nature to induce stepwise appropriate cell responses. It is hypothesized that any alteration in the cell membrane structure induces influx of Ca2+ ions. Ca2+ ions are required to activate degrading enzymes, such as phospholipases and lipoxygenases (LOX) that transform PUFAs bound to membrane phospholipids to lipidhydroperoxides (LOOHs). Enzymatic reduction products of LOOHs seem to serve as ligands of proteins, which induce gene activation to initiate a physiological response. Increasing external impact on cells is connected with deactivation of LOX, liberation of the iron ion in its active center followed by cleavage of LOOH molecules to LO * radicals. LO * radicals induce a second set of responses leading to generation of unsaturated aldehydic phospholipids and unsaturated epoxyhydroxy acids that contribute to induction of apoptosis. Finally peroxyl radicals are generated by attack of LO * radicals on phospholipids. The latter attack nearly all types of cell constituents: Amino- and hydroxyl groups are oxidized to carbonyl functions, sugars and proteins are cleaved, molecules containing double bonds such as unsaturated fatty acids or cholesterol suffer epoxidation. LOOH molecules and iron ions at the cell wall of an injured cell are in tight contact with phospholipids of neighboring cells and transfer to these reactive radicals. Thus, the damaging processes proceed and cause finally necrosis except the chain reaction is stopped by scavengers, such as glutathione. Consequently, PUFAs incorporated into phospholipids of the cell wall are apparently equally important for the fate of a single organism as the DNA in the nucleus for conservation of the species. This review intends to demonstrate the connection of cell alteration reactions with induction of lipid peroxidation (LPO) processes and their relation to inflammatory diseases, especially atherosclerosis and a possible involvement of food. Previously it was deduced that food rich in cholesterol and saturated fatty acids is atherogenic, while food rich in n-3 PUFAs was recognized to be protective against vascular diseases. These deductions are in contradiction to the fact that saturated fatty acids withstand oxidation while n-3 PUFAs are subjected to LPO like all other PUFAs. Considering the influence of minor food constituents a new theory about atherogenesis and the influence of n-3 PUFAs is represented that might resolve the contradictory results of feeding experiments and chemical experiences. Cholesterol-PUFA esters are minor constituents of mammalian derived food, but main components of low density lipoprotein (LDL). The PUFA part of these esters occasionally suffers oxidation by heating or storage of mammalian derived food. There are indications that these oxidized cholesterol esters are directly incorporated into lipoproteins and transferred via the LDL into endothelial cells where they induce damage and start the sequence of events outlined above. The deduction that consumption of n-3 PUFAs protects against vascular diseases is based on the observation that people living on a fish diet have a low incidence to be affected by vascular diseases. Fish are rich in n-3 PUFAs; thus, it was deduced that the protective properties of a fish diet are due to n-3 PUFAs. Fish, fish oils, and vegetables contain besides n-3 PUFAs as minor constituents furan fatty acids (F-acids). These are radical scavengers and are incorporated after consumption of these nutrients into human phospholipids, leading to the assumption that not n-3 PUFAs, but F-acids are responsible for the beneficial efficiency of a fish diet. PMID- 16270287 TI - MR-guided laser irradiation of recurrent glioblastomas. AB - We treated two patients with recurrent glioblastoma multiforme using Nd:YAG laser irradiation in the framework of a salvage therapy. The underlying concept is to achieve cytoreduction by partial coagulation of the tumor. Magnetic resonance imaging (MRI) follow-up examinations revealed a volume reduction of the laser irradiated areas, while the untreated parts of the tumor exhibited a progression. The survival time after the diagnosis of the recurrence was 16 and 20 months, respectively, which is substantially (about four times) longer than the natural history of the disease would suggest. In conclusion, cytoreduction by laser irradiation may be a promising option for patients suffering from recurrent glioblastoma multiforme. Future work should optimize the therapeutic regimen and evaluate this treatment approach in controlled clinical trials. PMID- 16270288 TI - Hemodynamic evaluation of the peripheral pulmonary circulation by cine phase contrast magnetic resonance imaging. AB - PURPOSE: To describe the normal flow patterns in peripheral pulmonary vessels with phase-contrast (PC) magnetic resonance imaging (MRI). MATERIALS AND METHODS: Twelve healthy adults (age = 33 +/- 7 years) underwent cine PC MRI of the segmental and central pulmonary arteries and veins by means of a breath-held segmented k-space technique. Flow patterns were analyzed on time-velocity curves and compared between the peripheral and central vessels. RESULTS: The pulsatile flow patterns in the segmental arteries and veins were similar among individuals. When compared with the central pulmonary arteries, the segmental arteries had a delay in the systolic and diastolic flow velocity waves, and an increased magnitude of the diastolic peaks, in relation to the systolic peaks. A prominent notch was present during the deceleration phase of the systolic flow velocity wave in 79% of the segmental arteries investigated. The segmental veins showed a typical pulmonary venous flow pattern, as seen in the central veins, with similar systolic-to-diastolic peak velocity ratios. CONCLUSION: Noninvasive evaluation of blood flow in intraparenchymal pulmonary vessels is feasible with PC MRI. This first description of normal flow patterns in segmental pulmonary arteries and veins can serve as basis for further investigation in the setting of altered pulmonary blood flows. PMID- 16270289 TI - Magnetic susceptibility effects on the accuracy of MR temperature monitoring by the proton resonance frequency method. AB - PURPOSE: To evaluate the error of MR temperature assessment based on the temperature-dependent Larmor frequency shift of water protons, which can result from susceptibility effects caused by the radiofrequency (RF) applicator. MATERIALS AND METHODS: Local frequency shifts due to RF applicator displacements were simulated numerically by means of a three-dimensional elementary dipole model. Experimental examinations using a water tank phantom equipped with a high precision screw thread were applied to examine temperature and movement effects for five commercially available, MR-compatible RF applicators. Measurements were performed at 1.5 Tesla. RESULTS: For single-needle electrodes perpendicular to the external field, a distortion of 0.1 ppm and 0.2 ppm was recorded at a distance of 17.5 mm and 12.5 mm, respectively, to the needle shaft. Cluster applicators and umbrella-shaped applicators caused distortions of 0.1 ppm up to distances of 36 mm. Sinusoidal dependence on applicator orientation was found with the highest values for perpendicular orientation and the lowest values for orientation parallel to the magnetic field. With a single electrode oriented perpendicular to the field at a distance of 1.5 cm and 2.0 cm, a needle displacement of 5 mm led to an error in temperature measurement of 16.3 degrees C and 7.5 degrees C, respectively. CONCLUSION: In MR temperature measurement, displacement of the RF applicator by patient movement or breathing leads to significant errors that have to be taken into account when PRF temperature maps are used to monitor tumor ablation in the presence of paramagnetic applicators. PMID- 16270290 TI - STIR vs. T1-weighted fat-suppressed gadolinium-enhanced MRI of bone marrow edema of the knee: computer-assisted quantitative comparison and influence of injected contrast media volume and acquisition parameters. AB - PURPOSE: To compare short tau inversion recovery (STIR) and T1-weighted (T1w) gadolinium (Gd)-enhanced fat-suppressed MRI of bone marrow edema (BME) of the knee, and investigate the influence of injected contrast media volume and variation of major acquisition parameters on apparent BME volume and signal contrast. MATERIALS AND METHODS: STIR and T1w Gd-enhanced fat-suppressed images were obtained from 30 patients with BME of the knee. Two groups of patients were examined with different MR scanners, acquisition parameters, and contrast media volumes. For both sequences, BME volume and signal contrast were assessed by computer-assisted quantification, and were compared through their arithmetic means and correlation coefficients (r(2)). The injected contrast media volume was also correlated with BME volume and signal contrast differences between sequences. RESULTS: A strong correlation between the STIR and Gd-enhanced T1w images was found for BME volume (r(2) = 0.96-0.99) and BME signal contrast (r(2) = 0.86-0.94). Despite the differences in MR acquisition parameters and injected contrast media volume, both sequences depicted an almost identical BME volume in both groups. Contrast media volume showed a moderate correlation (r(2) = 0.40) with BME volume differences. CONCLUSION: STIR is the optimum method for determining the size and signal contrast of BME. The injected contrast media volume appears to have only a limited influence on apparent BME volume. PMID- 16270291 TI - Optimization of indirect arthrography of the knee by application of external heat: initial experience. AB - PURPOSE: To examine the potential utility of applying heat to increase the uptake of intravenous gadolinium (Gd) contrast into the knee joint in order to optimize MR arthrography. MATERIALS AND METHODS: At 1.5T, 16 knees in eight patients without prior surgery, injury, or pain were examined before and 30 minutes after intravenous administration of Gd contrast (0.1 mM/kg). Between scans a heating pad was applied to the anterior aspect of eight randomly selected knees (the contralateral knee served as the control). Initial and postcontrast imaging consisted of identical axial T1-weighted sequences (TR/TE = 500/14 msec) without fat suppression. On the initial and postcontrast images, regions of interest (ROIs) were placed at identical locations in the suprapatellar pouch and the intercondylar notch by a reader blinded to the treated side. The values at these two locations were averaged and the change in joint signal intensity was calculated. The differences between the heated and unheated knees were also calculated. RESULTS: Seven of the eight knees treated with heat had increased joint enhancement compared to the contralateral control, with percentage changes in joint signal intensity (heated knee vs. control) of +38%, +80%, +121%, +145%, +150%, +164%, and +177%. Overall there was a doubling of signal intensity (125%) on the heated side compared to the contralateral control (with significance at P = 0.039). One patient was excluded because of a prior knee injury. CONCLUSION: The application of external heat increases uptake of intravenously administered Gd contrast into the knee joint, and may help to optimize indirect MR arthrography at a relatively low cost. PMID- 16270292 TI - Spontaneous craniospinal hypotension. AB - Spontaneous cerebrospinal fluid (CSF) leak has been called "spontaneous intracranial hypotension," emphasizing the intracranial symptoms and imaging findings. We present a patient with spontaneous CSF leak whose initial spinal magnetic resonance imaging (MRI) findings were thought to represent epidural tumor or infection. Subsequent MRI examinations showed an improvement of both intracranial and spinal CSF hypotension findings coinciding with clinical symptom resolution. We propose the term "spontaneous craniospinal hypotension" to better emphasize this syndrome's unifying intracranial and spinal pathophysiology and imaging findings. PMID- 16270293 TI - Influence of positional and angular variation of automatically planned short-axis stacks on quantification of left ventricular dimensions and function with cardiovascular magnetic resonance. AB - PURPOSE: To theoretically and experimentally investigate the influence of the automated cardiovascular magnetic resonance (CMR) scan planning pitfalls, namely inaccurate positioning and tilting of short-axis (SA) imaging planes, on quantification of the left ventricular (LV) dimensions and function. MATERIALS AND METHODS: Eleven healthy subjects and eight patients underwent CMR. Manually and automatically planned SA sets were acquired. To obtain the quantitative measurements of LV function, one observer performed image analysis twice. The agreement between planning methods, as well as the decomposition of the total variation into interstudy and intraobserver components was measured. RESULTS: The decomposition of the total variation showed that the interstudy factor accounts for 70-85% of the total variation, while the rest is due to the intraobserver factor. Moreover, the relative contribution of the interstudy factor remains independent from errors in slice positioning and small angular deviation of SA stacks from the optimal orientation. Good agreement between the theoretical and measured variability factors was observed. CONCLUSION: Global LV function derived from the automatically planned CMR acquisitions yield accurate quantification of the human cardiovascular system. Inaccurate positioning and tilting of SA images does not affect the quantitative measurements of LV function. The computer-aided system for automated CMR has proven clinical applicability. PMID- 16270294 TI - Value of contrast-enhanced, balanced cine-MR sequences in the assessment of apparent infarct size after acute myocardial infarction: a prospective comparison with delayed-enhancement sequences. AB - PURPOSE: To evaluate the diagnostic value of contrast-enhanced (CE) delayed cine steady-state free precession (SSFP) sequences in the assessment of apparent infarct size after acute myocardial infarction (MI). MATERIALS AND METHODS: Contrast-enhanced (CE) balanced cine-SSFP sequences were compared with delayed enhancement (DE) T1 sequences for their ability to detect segmental abnormal enhancement in 29 consecutive patients with recent successfully reperfused acute MI. RESULTS: The extent of myocardial involvement revealed by postcontrast cine SSFP sequences and DE images was closely correlated (Spearman r = 0.86, P < 0.001). There was no significant difference between the two sequences in assessing subendocardial or transmural involvement. CONCLUSION: In addition to DE sequences, CE cine-SSFP sequences should play a role in assessing necrotic and jeopardized myocardium after acute MI. PMID- 16270295 TI - Line-scan diffusion tensor MR imaging at 0.2 T: feasibility study. AB - PURPOSE: To investigate and measure apparent diffusion coefficient (ADC) and fractional anisotropy (FA) values using data obtained with line-scan diffusion weighted imaging (DWI) of human brains on a 0.2 Tesla MR imager. MATERIALS AND METHODS: Diffusion-tensor imaging (DTI) was performed on eight healthy volunteers. The signal-to-noise ratios (SNRs) of white matter and cerebrospinal fluid were measured. ADC and FA were also measured from the data obtained from all subjects. Three-dimensional corticospinal fiber tracts were reconstructed from the DT images and a qualitative evaluation was done. RESULTS: The total scan time was 52 minutes 30 seconds for 18 slices with full-tensor images covering the whole brain. The ADCs and FAs show the appropriate values, in comparison with values obtained at high field strength in previous studies. Corticospinal fibers were demonstrated more clearly on images obtained at 0.2 T than at 1.5T. CONCLUSION: DTI at low field strength may be feasible for clinical use to estimate the white matter of brain with limited coverage, which often may be sufficient. PMID- 16270296 TI - 3.0 T vs. 1.5 T MR angiography: in vitro comparison of intravascular stent artifacts. AB - PURPOSE: To evaluate the signal characteristics of different iliac artery stents in MR angiography (MRA) at 3 T in comparison with 1.5 T. MATERIALS AND METHODS: Sixteen iliac artery stents were implanted in plastic tubes filled with a solution of Gd-DTPA and imaged at 3 T and 1.5 T using a T1-weighted 3D spoiled gradient-echo sequence. Image analysis included a subjective assessment of artifact characteristics, signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) measurements in stented and unstented vessel parts, and quantitative measurements of total artifact size. RESULTS: The pattern of stent artifacts inside the stents evidently did not differ at 3 T and 1.5 T. The average total size of the artifact areas surrounding the stents was significantly larger at 3 T (P < 0.03). However, within the stented part of the vessel phantom, the signal of the lumen and its contrast to modeled surrounding tissue was significantly higher at the higher field. The mean SNR of the lumen increased from 95.5 at 1.5 T to 127.3 at 3 T, and the CNR of the vessel increased from 70.3 to 93. CONCLUSION: Assessment of the stent lumen in iliac artery stents in a phantom model is not compromised by imaging at 3 T compared to 1.5 T. The signal gain inside the stented part of the vessel lumen at higher field compensates for the higher degree of stent artifacts seen in stents made of steel or cobalt. PMID- 16270297 TI - The role of extracellular signal-regulated protein kinase in transcriptional regulation of the hypoxia marker carbonic anhydrase IX. AB - In the present study, we investigated the role of the extracellular signal regulated protein kinase (ERK) in regulation of the hypoxia marker, carbonic anhydrase IX (CAIX). U0126, a specific inhibitor of MEK1/2, downregulated CAIX expression induced by true hypoxia and cell density. CA9 promoter activity was similarly affected. Mapping of the U0126 effect revealed that both critical elements within the CA9 promoter, the hypoxia response element and the juxtaposed SP1-binding PR1, were inhibited. This confirmed that ERK signaling modulates CA9 promoter activity via its effects on hypoxia inducible factor-1 (HIF-1) and SP1. Further analysis of the U0126 effect on HIF-1-dependent transcription in MCF-7 cells identified p300, a transcriptional co-activator of HIF-1, as the target of ERK. Constitutively increased ERK activity in isogenic fibrosarcoma cell lines did not cause increased cell density-dependent CAIX expression/CA9 promoter activity. In HeLa cells, an inverse correlation between cell density-induced CAIX expression and ERK activation was observed: sparse cultures did not express CAIX and displayed high ERK activation, whereas CAIX expression in dense cultures was associated with low ERK activation. Collectively, our data do not support any quantitative relationship between ERK activation and CAIX expression. Thus, although ERK signaling is required for optimal CAIX expression, our data are consistent with a model in which constitutive basal ERK activity plays an auxiliary role in CA9 promoter transactivation by modulating activity of the transcription factor SP1 and the transcriptional co-activator p300. PMID- 16270298 TI - Oxygen gradients correlate with cell density and cell viability in engineered cardiac tissue. AB - For clinical utility, cardiac grafts should be thick and compact, and contain physiologic density of metabolically active, differentiated cells. This involves the need to control the levels of nutrients, and most critically oxygen, throughout the construct volume. Most culture systems involve diffusional transport within the constructs, a situation associated with gradients of oxygen concentration, cell density, cell viability, and function. The goal of our study was to measure diffusional gradients of oxygen in statically cultured cardiac constructs, and to correlate oxygen gradients to the spatial distributions of cell number and cell viability. Using microelectrodes, we measured oxygen distribution in a disc-shaped constructs (3.6 mm diameter, 1.8 mm thickness) based on neonatal rat cardiomyocytes cultured on collagen scaffolds for 16 days in static dishes. To rationalize experimental data, a mathematical model of oxygen distribution was derived as a function of cell density, viability, and spatial position within the construct. Oxygen concentration and cell viability decreased linearly and the live cell density decreased exponentially with the distance from the construct surface. Physiological density of live cells was present only within the first 128 microm of the construct thickness. Medium flow significantly increased oxygen concentration within the construct, correlating with the improved tissue properties observed for constructs cultured in convectively mixed bioreactors. PMID- 16270299 TI - Soluble adhesion molecules in young children with signs of beta-cell autoimmunity -prospective follow-up from birth. AB - BACKGROUND: This study aimed at evaluating the relationship between the circulating concentrations of soluble intercellular adhesion molecule-1 (sICAM-1) and sL-selectin and the appearance of beta-cell autoimmunity, and at assessing whether these molecules could assist in the identification of environmental factors implicated in the immune process damaging the pancreatic beta-cells. METHODS: Serum levels of soluble adhesion molecules were measured with enzyme linked immunosorbent assays over the first 2 years of life in 65 children seroconverting to positivity for autoantibodies and 65 control children, all with HLA-conferred susceptibility to type 1 diabetes (T1D). RESULTS: The total integrated concentrations of soluble adhesion molecules were comparable between the two groups. The autoantibody-positive children tended to have higher sL selectin concentrations during the 3-month seroconversion (SC) period than did the control children during the corresponding period (P = 0.07), the difference being significant (P = 0.03) after excluding subjects with signs of a concurrent enterovirus infection. Autoantibody-positive children had higher concentrations of sL-selectin in the 3-month period when an enterovirus infection was detectable than did the control children (P = 0.018). No significant difference could, however, be seen after excluding the children with concomitant seroconversion to autoantibody positivity. CONCLUSIONS: Elevated concentrations of sL-selectin are temporally associated with seroconversion to autoantibody positivity suggesting that leukocyte activation might coincide with the appearance of beta-cell autoimmunity. Early-onset progressive beta-cell autoimmunity, on the other hand, is not reflected in overall increased concentrations of soluble adhesion molecules in the peripheral circulation during the first 2 years of life in children carrying increased HLA-conferred disease susceptibility. Enterovirus infections (EVIs) are not independently associated with increased circulating sL selectin concentrations in young children with enhanced HLA-conferred susceptibility to T1D. PMID- 16270300 TI - Repeated antipsychotic drug exposure in developing rats: dopamine receptor effects. AB - Antipsychotic drugs are often prescribed to juvenile psychiatric patients, though their cerebral effects during development are incompletely described. Accordingly, we studied the effects of repeated treatment with dissimilar antipsychotic drugs on dopamine (DA) receptors in juvenile vs. adult rats. Tissue levels of DA receptor types (D1, D2, D3, and D4) in forebrain regions of juvenile rats were quantified after 3 weeks of daily treatment with representative first- (fluphenazine) and second-generation (clozapine and olanzapine) antipsychotics, and compared with similarly treated adult rats examined in previous studies. Fluphenazine, clozapine, and olanzapine all decreased D1 receptors in dorsolateral frontal and medial prefrontal cortex (MPC) of juvenile, but not adult rats. Conversely, all three test agents increased D2 labeling in MPC of adult, but not young animals. Fluphenazine and olanzapine, but not clozapine, also increased D2 receptor levels in hippocampus, and D4 levels in nucleus accumbens (NAc) and caudate-putamen (CPu) in both juvenile and adult brain. D3 receptors were not altered by any treatment in any brain region at either age. Only some DA receptor adaptations to antipsychotic treatment are shared by developing and mature animals. Developmental differences in DA receptor responses may account for differences in clinical effects of antipsychotic drugs between young and adult psychiatric patients. PMID- 16270301 TI - The influence of sex on extracellular dopamine and locomotor activity in C57BL/6J mice before and after acute cocaine challenge. AB - C57BL6/J (C57) mice serve as a useful animal model of cocaine abuse because they self-administer cocaine, exhibit place conditioning to cocaine, discriminate the interoceptive cues of cocaine, and are used for backcrossing strains of genetically modified mice. The present study was to examine the influence of sex on extracellular DA and locomotor activity in C57 mice in response to acute cocaine challenge. In the first experiment, male and female mice were implanted with guide cannulae aimed at the dorsal striatum. Microdialysates were collected in three consecutive phases: baseline, post-saline injection, and post-cocaine injection. Sex did not influence DA measurements during baseline or after intraperitoneal (i.p.) saline injection. Cocaine (20 mg/kg) injections increased peak extracellular DA of both sexes, and the increase was greater for males (278%+/-14.0%) than females (182.5%+/-10.8%) (P<0.05). In the second experiment, under conditions similar to the microdialysis experiment, locomotor activity of male and female mice was assessed during baseline, after saline injection, and after cocaine injection (5, 10, 20, or 40 mg/kg). Cocaine dose-dependently increased activity; however, sex did not influence locomotor activity during baseline, after saline, or after any cocaine dose. Results of the experiments established that cocaine (20 mg/kg) increased extracellular DA in the dorsal striatum to a greater extent in male than in female mice; however, when cocaine was administered under similar experimental conditions, sex did not influence cocaine stimulation of locomotor activity over a wide range of doses. PMID- 16270302 TI - Apolipoprotein E4 suppresses delayed-rectifier potassium channels in membrane patches excised from hippocampal neurons. AB - Recent studies show a clear association between Alzheimer's disease (AD) and the apolipoprotein E epsilon 4 allele (APOE4). The mechanisms underlying apoE4 mediated detrimental effects have not been well-clarified. The present study investigates possible effects of apoE4 on the delayed-rectifier potassium (IK) channels in inside-out membrane patches excised from rat hippocampal neurons. Acute application of apoE4 (0.5 microM) to the inside of the membrane patches markedly and reversibly suppressed the single IK channel activities. The average open probability and open frequency of IK channels decreased by (92.6+/-7.1)% and (88.6+/-3.2)%, respectively. The mean open time of IK channels decreased by (81.6+/-6.7)%, and the mean closed-time of them increased by 6.9+/-1.9 fold. Meanwhile, the mean current amplitude of IK channels was not significantly affected. In contrast, application of apolipoprotein A (apoA, 0.5 microM), another member of apolipoprotein family with similar molecular weight and amino acid sequence to apoE4, did not exhibit any effects on IK currents. These results indicate that apoE4 molecules can rapidly suppress the activities of IK channels in hippocampal neurons when they act on the inner side of the neuronal membrane. We propose that the overproduction of apoE4 in neurons may suppress normal IK channel activities and thus be responsible for the late-developed neuronal damages related to the pathogenesis of AD. PMID- 16270304 TI - Relationship between adiposity and body size reveals limitations of BMI. AB - The aims of this study were to assess 1) whether the stature-adjusted body mass index (BMI) is a valid proxy for adiposity across both athletic and nonathletic populations, and 2) whether skinfold measurements increase in proportion to body size, thus obeying the principle of geometric similarity. The research design was cross-sectional, allowing the relationship between skinfold calliper readings (at eight sites and between specific athletic and nonathletic groups, n = 478) and body size (either mass, stature, or both) to be explored both collectively, using proportional allometric MANCOVA, and individually (for each site) with follow-up ANCOVAs. Skinfolds increase at a much greater rate relative to body mass than that assumed by geometric similarity, but taller subjects had less rather than more adiposity, calling into question the use of the traditional skinfold-stature adjustment, 170.18/stature. The best body-size index reflective of skinfold measurements was a stature-adjusted body mass index similar to the BMI. However, sporting differences in skinfold thickness persisted, having controlled for differences in body size (approximate BMI) and age, with male strength- and speed trained athletes having significantly lower skinfolds (32% and 23%, respectively) compared with controls. Similarly, female strength athletes had 29% lower skinfold measurements compared to controls, having controlled for body size and age. These results cast serious doubts on the validity of BMI to represent adiposity accurately and its ability to differentiate between populations. These findings suggest a more valid (less biased) assessment of fatness will be obtained using surface anthropometry such as skinfolds taken by experienced practitioners following established procedures. PMID- 16270303 TI - Anatomic distribution of reinforcer selective cell firing in the core and shell of the nucleus accumbens. AB - We have previously reported that distinct populations of nucleus accumbens (NAc) neurons differentially encode information about goal-directed behaviors for "natural" (food/water) vs. cocaine reinforcement [J Neurosci 20:4255-4266, (2000)]. Here, the anatomic distribution of reinforcer-selective cell firing was examined within the core and shell of the NAc. Rats (n=8) were trained on a multiple schedule for water reinforcement and cocaine (0.33 mg/infusion) self administration. Next, microelectrode arrays (eight wires/array) were bilaterally positioned in the NAc core and shell, and cell firing was recorded during the multiple schedule. All electrode tip placements were then "marked," and histological reconstruction of each electrode position was completed. Of 93 NAc cells, 44 neurons (47%) exhibited 1 of 4 types of well-documented patterned discharges relative to the water- or cocaine-reinforced response. Of the 44 phasic cells, 39 neurons (89%) displayed differential, nonoverlapping neuronal firing patterns across the two reinforcer conditions (i.e., reinforcer-selective activity). Histological reconstruction of electrode placement revealed that NAc patterned discharges, specific to goal-directed behaviors for water or cocaine, were not limited to one NAc subregion but were evenly distributed and intermixed throughout the core and shell. These findings are discussed with respect to the functional organization of the NAc. PMID- 16270305 TI - Conduction velocity distribution in neurologically well-recovered but fatigued Guillain-Barre syndrome patients. AB - Many patients with Guillain-Barre syndrome (GBS) and chronic inflammatory demyelinating polyneuropathy (CIDP) suffer from excessive fatigue. To assess whether this fatigue might be related to changes in slow-conducting nerve fibers, we determined the conduction velocity distribution (CVD) in the median nerve. Thirteen fatigued but neurologically well-recovered GBS patients, 2 fatigued and stable CIDP patients, and 19 healthy controls participated in this study. Conventional maximal nerve conduction velocities (NCVs) did not show differences between GBS patients and healthy controls. However, in both GBS and CIDP patients the CVD was altered, showing significant narrowing of the velocity distribution with loss of the fastest- and slowest-conducting fibers. These changes were most pronounced in the subgroup of patients with the lowest fatigue scores. We therefore conclude that the observed CVD changes in patients are not likely to contribute to persisting complaints of fatigue after GBS. PMID- 16270306 TI - Effects of changing wrist positions on finger flexor hypertonia in stroke survivors. AB - We sought to establish whether spastic hypertonia results from changes in intrinsic muscle properties or from altered stretch reflex properties. We hypothesized that finger flexor spastic hypertonia is primarily of neural origin, and that the dynamics of spastic muscle responses to stretch should therefore reflect the dynamics of muscle spindle receptor responses. In 12 stroke survivors, we recorded torque and electromyographic (EMG) responses of extrinsic finger flexors to constant-velocity rotation of the metacarpophalangeal (MCP) joints of the affected hand, over a range of initial muscle lengths. Stretch velocity was set to 6 degrees, 50 degrees, 150 degrees, or 300 degrees per second. Muscle length changes were imposed by changing wrist angle between 0 degree, 25 degrees, and 50 degrees of flexion. We found that reflex torque and EMG responses exhibited both velocity and length dependence, and there were significant interactions between velocity and length, replicating known characteristics of muscle spindle receptors. Our results support the hypothesis that finger flexor hypertonia is primarily of neural origin, and that it accurately reflects spindle receptor firing properties. PMID- 16270307 TI - Evidence for MHC I-restricted CD8+ T-cell-mediated immunopathology in canine masticatory muscle myositis and polymyositis. AB - Masticatory muscle myositis (MMM) is the most common inflammatory myopathy (IM) in dogs, associated with antibodies against myosin. To further elucidate the immunopathogenesis, we investigated muscles of 53 dogs with MMM, 32 dogs with polymyositis (PM), and 4 dogs suffering from both, with regard to the presence and location of CD4(+) and CD8(+)T cells, B cells, macrophages, major histocompatibility complex (MHC) class I and class II antigens, and autoantibodies. CD8(+)T cells were found in MMM (91%) and PM (75%), mostly paralleled (68% and 61%) by enhanced expression of MHC class I antigen on muscle fibers. CD8(+)T cells invading intact and neighboring necrotic muscle fibers were present in MMM (39%) and PM (42%). Dogs with MMM lacking intramuscular (26%) and circulating (36%) autoantibodies also had CD8(+) T-cell infiltrations and muscle fiber lesions. Since MHC class I antigen and CD8(+) T cells were detected in the presence of CD4(+) T cells, regardless of antimuscular antibodies, we consider MMM and PM in the dog as a CD8(+) T-cell-mediated immunopathological disease that initiates muscle-fiber destruction and leads to production of myosin autoantibodies. PMID- 16270308 TI - Carbohydrate mimicry of Campylobacter jejuni lipooligosaccharide is critical for the induction of anti-GM1 antibody and neuropathy. AB - The expression of ganglioside-mimicking structures of Campylobacter jejuni lipooligosaccharides (LOS) is considered essential for the induction of antiganglioside antibodies that lead to Guillain-Barre syndrome (GBS). The galE gene in C. jejuni is involved in the biosynthesis of the LOS outer-core oligosaccharide structures. We have demonstrated that the C. jejuni HB9313 (HS:19) parental strain expresses a LOS structure containing GM1-like epitopes, and the C. jejuni knockout mutant of the galE gene expresses a truncated LOS structure without GM1-like epitopes. To clarify whether the ganglioside-like structures in Campylobacteri LOS are crucial for induction of antiganglioside antibody responses and neuropathy, we performed immunization experiments in guinea pig models using the parental strain HB9313 and its galE mutant derivative. The anti-GM1 IgG antibody responses in immunized animals were measured by enzyme-linked immunosorbent assay. Sciatic nerve specimens were evaluated pathologically. High levels of the anti-GM1 IgG antibody were induced in guinea pigs immunized with HB9313, but not in those immunized with the galE mutant. The mean percentage of abnormality of sciatic-nerve teased fibers from animals sensitized with C. jejuni HB9313 was significantly higher than from animals immunized with the galE mutant. Furthermore, significant changes were found in semithin sections of the sciatic nerve from animals inoculated with C. jejuni HB9313. The major pathological finding was axonal degeneration; no significant morphological findings, except for occasional demyelination, were observed in animals immunized with the galE mutant. These results indicate that ganglioside-mimicry structures in C. jejuni LOS are necessary for induction of antiganglioside antibody response and neuropathy. PMID- 16270309 TI - Morphological changes in the clasper gland of the Atlantic stingray, Dasyatis sabina, associated with the seasonal reproductive cycle. AB - The clasper gland of the Atlantic stingray, Dasyatis sabina, was examined over a 1-year period, covering an entire reproductive cycle. Changes in clasper gland tissue architecture, fluid production, and cell proliferation were assessed. No changes in tissue architecture were observed. Evidence of cell proliferation in the gland epithelium was not detected using immunocytochemistry for proliferating cell nuclear antigen, a cellular marker of mitosis. Epithelial cells were not observed to undergo mitosis, and cell membranes remained intact. The lack of structural changes and epithelial cell proliferation supports the proposed merocrinal mode of fluid secretion. Rays captured in nonbreeding months had clasper glands that exhibited tubules with reduced lumens. In contrast, rays caught during the breeding season had clasper gland tubules with enlarged lumens. Clasper gland fluid production was quantified through measurements of the fluid area and tubule area calculated from digital images. Clasper gland fluid production was significantly higher during the mating period than during months not associated with copulatory activity. These data support the notion that the clasper gland is involved in stingray copulatory activity. This study adds to the limited amount of literature focused on this poorly understood component of reproduction in skates and rays. PMID- 16270310 TI - Friendships and relationships: Sacramento AANCART's best practice. AB - The best practice of Sacramento's Asian American Network for Cancer Awareness Research and Training program is friendships and relationships. PMID- 16270311 TI - Cancer and social justice: a demographic, economic, historic, sociocultural, and ethical perspective. AB - In the current presentation, as a first-generation Asian-American immigrant, the author discussed the dire inequities of the current cancer prevention and control systems in the U.S. and attempted to analyze the root causes of the problem. The universal concern is that the occurrence of cancer, cancer's behavioral antecedents, (diet, physical activity, and tobacco use), the early detection of cancer, and cancer survivorship all relate inversely to education, income, social class, and white race. In other words, not only are cancer rates higher among lesser educated, poorer, and socially deprived individuals, but the availability and benefits of primary, secondary, and tertiary cancer prevention also are rationed, consciously or sub-consciously, by current society within and outside the borders of the U.S. Asian Americans are one of the unrecognized populations among these deprived groups. The objective of this article was to provide a thoughtful perspective on this very real problem and why it persists. Because of the audience at the Asian American Network for Cancer Awareness, Research, and Training meeting, where the current report was presented, the author tried to avoid a treatise on Asian philosophy and values but could not resist the comment that, in archaic Chinese terms, the public health and health care systems in the U.S. today lack balance and harmony. PMID- 16270312 TI - Quality of life in Chinese patients with breast cancer. AB - Chinese are the largest Asian group in the U.S., constituting 23.8% of the nation's total Asian-American population. Cancer is the leading cause of death for female Asian Americans, and breast cancer is the most frequently diagnosed cancer among females for all racial/ethic groups in San Francisco, which ranks 4th in the number of Asian Americans and where 152,620 Chinese account for 19.6% of the city's total population. Previous observations among Chinese immigrant women suggested that a diagnosis of breast cancer may be more detrimental to their well being compared with Chinese women who are born and raised in the U.S. This difference may be due to the lower socioeconomic status, limited English proficiency, poor understanding of the Western medical system among immigrants or other educational or financial considerations. In this qualitative pilot study, the authors sought to increase understanding of the relation between cultural beliefs and quality of life (QOL) among immigrant Chinese women with breast cancer in San Francisco. Specific objectives were 1) to identify these patients' beliefs regarding cancer, life expectancy, and discussion of advance directives; 2) to explore how these beliefs relate to patient QOL; and 3) to generate hypotheses for further study. The overall objective of the pilot study was to investigate questions central to the QOL issue, including what defines QOL for immigrant women and how QOL for them is similar to or different from that for American-born Chinese women. PMID- 16270313 TI - Cancer health disparities among Asian Americans: what we do and what we need to do. AB - Asian Americans are the nation's fastest growing racial group in terms of percentages, and they constitute a very heterogeneous population. The author reviewed the literature and proposed an agenda to reduce cancer health disparities based on this review and the accomplishments and aspirations of the National Cancer Institute-funded Asian American Network for Cancer Awareness, Research, and Training. The Asian American cancer burden is unique, unusual, and, to a certain extent, unnecessary. The Asian American cancer burden is unique, because Asians are the only racial/ethnic population to experience cancer as the leading cause of death. The unusual aspects of the cancer burden among Asian Americans include experiencing proportionally more cancers of infectious origin, such as human papillomavirus-induced cervical cancer, hepatitis B virus-induced liver cancer, and stomach cancer, than any other racial/ethnic population and, at the same time, experiencing an increasing numbers of cancers associated with "Westernization." To a certain extent, the cancer burden for Asian Americans is unnecessary if barriers to cancer screening, overcoming resistance to physician visits, and culturally competent interventions to reduce smoking, unhealthy diet, and increasing proper exercise can be instituted. Reducing cancer health disparities among Asian Americans will involve research into their unique, unusual, and unnecessary cancer burden. PMID- 16270314 TI - Development of an English as a second language curriculum for hepatitis B virus testing in Chinese Americans. AB - Chinese Americans are at disproportionately high risk of liver cancer. A major risk factor for liver cancer in Asia is infection with hepatitis B virus (HBV): Approximately 80% of liver cancers are linked to HBV, and chronic carriers of HBV are > 100 times more likely to develop liver cancer compared with noncarriers. However, many adults, particularly those who have immigrated to the U.S., remain untested and therefore unvaccinated or unmonitored for the disease. Chinese Americans are mostly foreign born, and more recent arrivals face multiple social and health challenges. Many require special attention from public health professionals because of low levels of acculturation and difficulties learning English. It has long been established that an English as a Second Language (ESL) curriculum can teach immigrant adults and their family's important life skills, such as job training and citizenship. The authors report on their plans to develop and pilot test a culturally appropriate curriculum that will motivate Chinese ESL students to obtain a blood test for the detection of the HBV. PMID- 16270315 TI - Brain and sensory organ morphology in Antarctic eelpouts (Perciformes: Zoarcidae: Lycodinae). AB - Eelpouts of the family Zoarcidae comprise a monophyletic group of marine fishes with a worldwide distribution. Centers of high zoarcid diversity occur in the North Atlantic and North Pacific, with important radiations into the Arctic, along southern South America, and into the Southern Ocean around Antarctica. Along with snailfishes (Liparidae), zoarcids form an important component of the non-notothenioid fauna in the subzero shelf waters of Antarctica. We document the anatomy and histology of the brains, cranial nerves, olfactory apparatus, cephalic lateral lines, taste buds, and retinas of three Antarctic zoarcid species, living at depths of 310-939 m, representing three of the nine genera from this region. The primary emphasis is on Ophthalmolycus amberensis, and we provide a detailed drawing of the brain and cranial nerves of this species. Although this brain reflects general perciform neural morphology, it exhibits a reduction of the (optic) tecta and the eminentia granulares and crista cerebellares of the lateral line system. Interspecific differences among the three species are slight. The olfactory rosette consists of three to four lamellae and the nasal sac, contrary to the claim of Fanta et al. ([2001] Antarct Rec, Natl Inst Polar Res, Tokyo 45:27-42), is not in communication with the cephalic lateral line system. Primary olfactory neurons are abundant and converge on branches of the olfactory nerve. Numerous taste buds are located in the lips. All three species lack an ocular choroid rete and have relatively thin retinas with a low cell density and a single bank of rods as the only type of photoreceptor. Neural diversification among Antarctic zoarcids has not involved the evolution of sensory specialists; brain and sensory organ morphologies do not approach the condition seen in primary deep-sea fishes, or even that of some sympatric non-perciform secondary deep-sea fishes, including liparids and muraenolepidids (eel cods). There may be phylogenetic constraints on brain morphology in perciforms such that we do not see extreme specialization in sensory and neural systems for deep habitats. We suggest that the brains and sensory organs of Antarctic zoarcids reflect habitation of 500-2,000-m depths and likely reflect morphologies seen in zoarcids living on continental slopes elsewhere in the world. This balance among the sensory modalities makes zoarcids relatively generalized among secondary deep-sea fishes and may be one of the reasons this opportunistic and adaptable group has been successful in colonizing a variety of emergent and ephemeral habitats. PMID- 16270316 TI - Salvage temozolomide for prior temozolomide responders. AB - BACKGROUND: Temozolomide (TMZ) often is used as adjuvant or first-line therapy for patients with glioma. Because of potential hematologic complications, it usually is discontinued after 12-18 cycles, even in responders. Subsequent salvage therapies are reported to have limited efficacy at the time of disease recurrence. In the current study, the authors assessed the outcome and complications of reusing TMZ at the time of disease recurrence in patients who previously responded to treatment. METHODS: A retrospective review of patients with recurrent/progressive glioma who had a history of response to TMZ and were treated with the same agent at the time of disease recurrence was conducted. RESULTS: Fourteen patients were identified (8 men and 6 women). The median age of the patients was 56 years (range, 25-67 yrs) at the time of diagnosis; 9 patients had glioblastoma, 3 had anaplastic astrocytoma, and 2 patients had low-grade oligodendroglioma. No patient developed disease progression while receiving the initial TMZ treatment. At the time of the initial disease recurrence, 13 patients were readministered TMZ. One patient received TMZ at the time of second disease recurrence. All patients were assessed for radiographic response. Objective response or stable disease was achieved in 6 patients (43%; 95% confidence interval [95% CI], 21-67%) and the 6-month progression-free survival was 36% (95% CI, 16-61%). CONCLUSIONS: TMZ was found to be well tolerated and effective in this setting, suggesting that repeat use of TMZ in previous responders warrants further investigation. PMID- 16270317 TI - A comparison and evaluation of the predictions of relational and conjunctive accounts of hippocampal function. AB - Relational and conjunctive memory theory each postulate that the hippocampus participates in the formation of long-term memory representations comprised of associations between multiple elements. The goals of the current work were to clarify and contrast these theories by outlining the nature of the representations that are spared vs. impaired following hippocampal damage according to each theoretical perspective. Relational theory predicts that hippocampal lesions will impair performance on tasks that require the formation of new long-term representations in which distinct elements must be regarded in relation to all other elements. Representations that remain intact despite hippocampal damage include separate representations of distinct individual elements or multiple stimuli fused into a static "blend" such as several elements viewed from one vantage point. Additionally, the relational account predicts that rapid incidental online processing of the relations can be achieved through structures other than the hippocampus, but this information will not be stored. In contrast, conjunctive theory predicts that hippocampal damage will impair the rapid formation of unitary representations that contain features of elements and their relative relationships bound in an inflexible manner. Deficits in the rapid formation of these conjunctive representations result in impaired performance on tasks that require rapid incidental stimulus binding. However, intact formation of conjunctive representations can occur over multiple trials in the service of problem solving. Using these theoretical frameworks, recent findings from the human and nonhuman animal literature are reexamined in order to determine whether one theory better accounts for current findings. We discuss empirical studies that serve as "critical experiments" in addressing the relational vs. conjunctive debate, and find that the predictions of relational theory are supported by existing findings over those from the conjunctive account. PMID- 16270318 TI - Adjuvant systemic therapy for male breast carcinoma. AB - BACKGROUND: In the current study, the authors describe the M. D. Anderson experience with adjuvant systemic therapy in male breast carcinoma patients. METHODS: A total of 156 men with a diagnosis of breast carcinoma registered and were treated at the M. D. Anderson Cancer Center between 1944 and 2001. One hundred thirty-five men had nonmetastatic breast carcinoma at diagnosis and were included in this analysis. Patients' charts were retrospectively reviewed to obtain details of patient characteristics, adjuvant therapy, and outcomes. Analysis was performed with descriptive statistics; the log rank test was used to compare outcomes. RESULTS: The median patient age was 59 years (range, 25-80 yrs). Median follow-up was 13.8 years (range, 0.6-32.5 yrs). Sixty percent of patients had tumors 2 cm or smaller. Pathologic lymph node involvement was seen in 55% of patients. Tumors were estrogen receptor-positive in 85% of cases and progesterone receptor-positive in 71%. Chemotherapy was administered to 32 men (84% with adjuvant chemotherapy, 6% with neoadjuvant chemotherapy, and 9% with both). Approximately 81% received anthracycline-based regimens; 9% received additional taxanes; and 16% were treated with cyclophosphamide, methotrexate, and 5-fluorouracil (CMF). The median number of cycles was 6 (range, 4-14 cycles). Thirty-eight men received adjuvant hormonal therapy (92% received tamoxifen and 8% were treated with other therapy). The 5-year and 10-year overall survival rates were 86% and 75%, respectively, for men with lymph node-negative disease and 70% and 43%, respectively, for men with lymph node-positive disease. For men with lymph node-positive disease, adjuvant chemotherapy was associated with a lower risk of death (hazards ratio [HR] of 0.78), although this difference was not statistically significant. Overall survival was significantly better for men who received adjuvant hormonal therapy (HR of 0.45; P = 0.01). CONCLUSIONS: This relatively large series of men with breast carcinoma suggests that men benefit from adjuvant systemic therapy for breast carcinoma, with the greatest benefit from adjuvant hormonal therapy. PMID- 16270319 TI - Preoperative lymph node staging of early-stage cervical carcinoma by [18F]-fluoro 2-deoxy-D-glucose-positron emission tomography. AB - BACKGROUND: Increasing evidence has documented the value of positron emission tomography (PET) in oncology, but only limited data are available comparing PET findings with the pathologic status of regional lymph nodes in patients with cervical carcinoma. The objective of this study was to determine the sensitivity and specificity of PET in detecting lymph node metastasis in women with early stage cervical carcinoma. METHODS: The authors performed a retrospective review of all patients with Stage IA-IIA cervical carcinoma who underwent PET before surgery from 1999 to 2004. The status of the regional lymph nodes was correlated with lymph node pathology. RESULTS: Fifty-nine patients were identified. Pelvic lymph node metastases were present in 32% of the patients and were detected by PET with a sensitivity of 53%, a specificity of 90%, a positive predictive value (PPV) of 71%, and a negative predictive value (NPV) of 80%. Paraaortic lymph node disease was present in 9% of patients and was detected by PET with a sensitivity of 25%, a specificity 98%, a PPV of 50%, and an NPV of 93%. The mean size of the tumor deposits was larger in the PET-positive pelvic nodes (15.2 mm; range, 2-35 mm) than in the PET-negative lymph nodes (7.3 mm; range, 0.3-20 mm; P = 0.002). Computed tomography (CT) scans were obtained before surgery in 42 patients. The combined sensitivity of PET and CT in these patients was 75%. PET alone detected 9 (36%) of the positive lymph node groups, whereas CT alone detected 3 (12%) of the positive lymph node groups. Neither PET nor CT detected the positive lymph node groups in 8 patients (32%). CONCLUSIONS: Pathologic validation of PET imaging demonstrated a low sensitivity and a high specificity for PET in patients with early-stage cervical carcinoma. PMID- 16270320 TI - Zoledronic acid suppresses lung metastases and prolongs overall survival of osteosarcoma-bearing mice. AB - BACKGROUND: Although there is no doubt that bisphosphonates (BPs), specific inhibitors of osteoclasts, are beneficial for the treatment of bone metastases, their effects on visceral metastases are unclear. The effect of zoledronic acid (ZOL) was examined in vivo on lung metastasis progression and animal survival, and in vitro on the cellular mechanisms involved. METHODS: An animal model of lung metastasis was developed in C3H/He mice inoculated intravenously with a spontaneous murine osteosarcoma POS-1 cell line. Lung metastasis was determined at the time of autopsy. ZOL was assessed in vitro on POS-1 cell proliferation, cell cycle progression, and caspase-1 and -3 activities. RESULTS: The overall survival in five independent experiments (two series treated with ZOL 0.1 mg/kg twice a week, and three series with 0.1 mg/kg five times a week) showed a significant increase of the actuarial survival: 0.422 +/- 0.07 in ZOL-treated animals versus 0.167 +/- 0.07 in controls (P = 0.036). Lung metastases were absent in all ZOL-treated mice that survived more than 21 days postinjection as revealed by macroscopic and histologic analysis. In vitro, a 48-hour incubation with 10 microM ZOL inhibited POS-1 cell line proliferation associated with cell cycle arrest in S-phase. In addition, ZOL induced a weak increase of caspase-3 activity, but not caspase-1. CONCLUSION: We demonstrate that ZOL exerts a direct antitumor effect on POS-1 cells in vitro, significantly diminishes osteosarcoma induced lung metastasis in vivo, thereby prolonging survival of POS-1-inoculated animals. PMID- 16270321 TI - Five genes from chromosomal band 8p22 are significantly down-regulated in ovarian carcinoma: N33 and EFA6R have a potential impact on overall survival. AB - BACKGROUND: Loss of heterozygosity on chromosomal band 8p22 is a common event in several epithelial tumors including ovarian carcinoma. So far, no clear evidence for a tumor suppressor gene (TSG) in this region has been found. METHODS: On the basis of publicly available expression data in ovarian tissues, the authors selected the eight most noteworthy genes from 8p22 (DLC1, N33, ZDHHC2, FLJ32642, PDGFRL, MTSG1, PCM1, and EFA6R) for a detailed expression analysis in 58 primary ovarian carcinoma tissues and in 38 ovarian cancer cell lines by using quantitative real-time reverse transcriptase-polymerase chain reaction (qRT-PCR). Expression data were correlated to various clinicopathologic characteristics and survival. RESULTS: Two genes showed a significantly (P< 0.05) lower expression in grade 3 tumors compared with tumors of lower grade (N33) or compared with normal controls and tumors with lower grade (EFA6R). Expression of N33 and EFA6R seems to have an impact on survival, in particular when the combined expression of both genes was used as predictive factor (P< 0.003). In addition, N33 and EFA6R showed a complete loss of expression in several ovarian cancer cell lines. Three genes (FLJ32642, MTSG1, and PCM1) had a significantly (P< 0.001, P< 0.004, and P< 0.001) lower expression in primary ovarian carcinoma compared with controls (ovarian tissues and cysts). CONCLUSIONS: Two to five new potential tumor suppressor or antagonizing gene candidates (N33 and EFA6R with impact on survival, and potentially FLJ32642, MTSG1, and PCM1) for ovarian carcinoma, were identified from the chromosomal band 8p22 and are promising candidates for further functional analysis in ovarian carcinoma. PMID- 16270322 TI - Medial septal modulation of the ascending brainstem hippocampal synchronizing pathways in the anesthetized rat. AB - Independent and combined electrical stimulation pairings of the medial septum (MS), posterior hypothalamus (PH), and reticular pontine oralis (RPO) of the brainstem were performed in the acute urethane anesthetized rat, while recording field activity from electrodes in either the stratum oriens or stratum moleculare of the hippocampal formation. Theta frequency and power were measured during independent stimulation of each nuclei and during combined stimulation using three pairings: (1) MS-PH (2) MS-RPO and (3) PH-RPO. Each pairing consisted of parameters known to elicit theta of a high frequency for one nucleus, and parameters known to elicit a low frequency for the second nucleus. This methodology allowed us to observe whether one nucleus preferentially modulated theta activity in the hippocampus in terms of frequency and power. The MS was observed to reset theta frequency in both the upward and downward direction when stimulated in combination with either the PH (Experiment 1) or the RPO (Experiment 2). In Experiment 3 (PH-RPO), the structure receiving the higher intensity stimulation had the predominate effect on theta frequency. With MS stimulation combinations, the power of the elicited theta activity was found to increase over the independent stimulation in some cases during Experiment 1. Likewise, in Experiment 2, the combined stimulation produced a power that in most cases was significantly greater than that measured during the independent stimulations. This effect was not observed with PH and RPO stimulation combinations. The combined stimulation of the PH and RPO yielded a power similar to the independent PH stimulations. The findings support the following conclusions: (1) the major theta generating activity of the ascending brainstem synchronizing pathways involves projections from the RPO to the PH, relayed through the MS, to the hippocampal formation; and (2) that the MS directly controls theta amplitude and secondarily translates the level of ascending brainstem activity into the appropriate frequency of hippocampal theta. PMID- 16270323 TI - Houston AANCART best practices: from vision to synergy to reality. AB - The theme for the 2004 AANCART Academy, "Community Partnerships for Cancer Control: From Vision to Synergy to Reality", characterizes Best Practices for the Houston AANCART site. Researchers and community members share a common vision for addressing the cancer and health disparities that exist in our Asian community. They banded together synergistically to bring to reality the programs and projects that are enabling more Asian Americans to understand their risks for cancer, receive screening and education, and access treatment and survivorship support. Along the way, Houston AANCART was also able to conduct meaningful and relevant community-based participatory research and to train young Asian and other investigators in how to reach out to this community in a culturally sensitive and appropriate manner. PMID- 16270324 TI - Highlights/best practices of San Francisco's Asian American Network for Cancer Awareness, Research, and Training (AANCART). AB - The Asian American Network for Cancer Awareness, Research, and Training in San Francisco (AANCART-SF) consists of two distinct entities, working in cooperation to advance cancer awareness, research, and training among Asian Americans: a university-based group with expertise in the Vietnamese community and a community based health plan with expertise in the Chinese community. In addition to the goals shared with other AANCART sites, AANCART-SF is a unique effort in capacity building in that it aims to expand and export community-academic research expertise from one Asian population, the Vietnamese, to other Asian populations. It also aims to build the research capability of those serving the Chinese community through a health plan. PMID- 16270325 TI - Medial septal modulation of the ascending brainstem hippocampal synchronizing pathways in the freely moving rat. AB - Rats implanted with hippocampal recording electrodes were tested in a wheel running apparatus under three conditions: (1) independent electrical stimulation of the medial septal nucleus (MS); (2) independent electrical stimulation of the posterior hypothalamic nucleus (PH); and (3) combined electrical stimulation of the MS and PH using pairings of two stimulation conditions, 7 or 10 Hz stimulation of the MS, and a low- or high-intensity PH stimulation. Quantitative measures of running speed were taken, and hippocampal recordings were subjected to fast-Fourier transform analysis. Electrical stimulation of the PH induced wheel-running behavior; running speed and the accompanying hippocampus (HPC) theta frequency increased with increase in stimulation intensity. Electrical stimulation of the MS failed to induce wheel-running behavior despite the fact that HPC theta was induced at the frequency of the applied stimulation (7 and 10 Hz). Electrical stimulation of the MS reset the frequency of HPC theta induced by PH stimulation in both the upward and downward directions and increased theta power, while wheel-running speed was modulated in a downward direction only. PMID- 16270326 TI - 1999-2001 Cancer mortality rates for Asian and Pacific Islander ethnic groups with comparisons to their 1988-1992 rates. AB - We report upper and lower boundary estimates of the 1999-2001 site-specific cancer mortality rates for Asian Indians, Chinese, Filipinos, Koreans, Vietnamese, Native Hawaiians, and Samoans. These rates are for the seven states (California, Hawaii, Illinois, New Jersey, New York, Texas, and Washington) that officially record mortality data for these ethnicities. The rates are based on the 2000 Census, which reports two population counts as follows: persons who identify themselves as belonging to a single ethnic group (which forms the basis for an upper boundary estimate of the rates) and persons who identify themselves as belonging to a single ethnic group or to multiple groups that include the single ethnic group (which forms the basis for a lower boundary estimate for the rates). The top five cancers for each Asian and Pacific Islander ethnic group by gender are reported. In addition, the 1988-1992 cancer mortality rates based on the 1990 Census for Chinese, Filipino, Japanese, and Native Hawaiians are determined. Their 1999-2001 and 1988-1992 rates are compared. PMID- 16270327 TI - Protein oxidation by chronic pulmonary diseases in children. AB - The oxidation of proteins may play an important role in the pathogenesis of chronic inflammatory lung diseases, and may contribute to lung damage. However, the extent of oxidation and the distribution among proteins are not known for most pediatric lung diseases. In this work, protein oxidation was assessed as protein carbonyls. Bronchoalveolar lavages (BAL) from children with chronic lung diseases were investigated by dot-blot assay for content and for pattern of distribution of oxidized proteins by two-dimensional (2D) electrophoresis and Western blotting. Significantly higher levels of protein oxidation than in healthy controls were determined in groups of patients with interstitial lung disease, gastro-esophageal reflux disease, and pulmonary alveolar proteinosis. The proteins most sensitive to oxidation were serum albumin, surfactant protein A, and alpha1-antitrypsin. Our data show increased oxidative stress in lungs of children with chronic pulmonary diseases, with significant interindividual variations. The extent of protein oxidation was proportional to the count of neutrophilic granulocytes in BAL fluid. These findings strongly support the concept that an abundance of reactive oxygen species produced during neutrophilic inflammation may be a deleterious factor, leading to pulmonary damage in these patients. PMID- 16270328 TI - In vivo carbon-13 magnetization transfer effect. Detection of aspartate aminotransferase reaction. AB - One of the most remarkable achievements of in vivo NMR spectroscopy has been the detection of rapid enzyme-catalyzed exchange reactions using phosphorus-31 magnetic resonance spectroscopy-based magnetization transfer experiments. In this paper, we report, for the first time, the in vivo carbon magnetization transfer (CMT) effect and in vivo detection of the CMT effects of the alpha-ketoglutarate <--> glutamate and the oxaloacetate <--> aspartate reactions, both of which are catalyzed by aspartate aminotransferase. By saturating the carbonyl carbon of alpha-ketoglutarate at 206 ppm in alpha-chloralose anesthetized adult rat brain, the unidirectional glutamate --> alpha-ketoglutarate flux was determined to be 78 +/- 9 mumol/g/min (mean +/- SD, n = 11) following i.v. infusion of [1,6 (13)C(2)]D-glucose. Contribution from aspartate aminotransferase-catalyzed partial reactions to the observed CMT effects was emphasized. Because of the large chemical shift separation between the alpha-carbons of the amino acids and the carbonyl carbons of the corresponding cognate keto acids, the spillover of the saturation radiofrequency pulses to the alpha-carbon resonances was negligible. The results indicate that the magnetization transfer effects of aspartate aminotransferase-catalyzed reactions can be used as new biomarkers accessible to non-invasive in vivo magnetic resonance spectroscopy techniques. PMID- 16270329 TI - Effect of strong homonuclear proton coupling on localized (13)C detection using PRESS. AB - The effect of strong homonuclear proton coupling on (13)C incorporation measurements by either indirect or direct means was investigated (and illustrated with glutamate) both numerically and experimentally at 3.0 T. In particular, two sequences were considered, each using a proton PRESS sequence for localization. The indirect (13)C detection method incorporated the POCE (proton observe carbon edited) technique onto PRESS, and for direct (13)C detection a DEPT (distortionless enhancement by polarization transfer) sequence was appended to the PRESS localization. Both analysis and experiment demonstrate that when strong homonuclear coupling of protons is additional to heteronuclear coupling with (13)C spins, the (13)C measures derived from either the indirect PRESS-POCE sequence or the direct-but-enhanced PRESS-DEPT sequence are significantly modified. Specifically, the MR lineshapes of both (13)C-bonded and nonbonded protons are changed during (13)C incorporation, giving rise, for example, to a potential cross-contamination of < or =30% between glutamate (13)C(3) and (13)C(4) measures from the PRESS-POCE indirect method. During direct-but-enhanced detection, the DEPT enhancement is reduced for glutamate (13)C(2), (13)C(3), and (13)C(4) but not equally, and the reduction is further exacerbated by proton PRESS localization, which gives rise to enhancements that are strong functions of PRESS TE(1) and TE(2). PMID- 16270330 TI - Three-dimensional reconstruction of limited-view projections for contrast enhanced magnetic resonance angiography at high temporal and spatial resolution. AB - The feasibility of reconstructing three-dimensional (3D) MRI data sets from limited-view projections is investigated in phantom and in vivo animal studies to improve the temporal resolution of magnetic resonance angiography without sacrificing spatial resolution. Thirty-two pairs of orthogonal biplane projections are acquired in an interleaved manner during the first pass of a contrast agent. The full data set is reconstructed as a priori 3D information. Each pair of projections is then reconstructed into an individual 3D data set based on a correlation analysis with the a priori data set. In this way, time resolved 3D data sets at 1- to 2-s time intervals are reconstructed with submillimeter spatial resolution. Artifacts are limited if the image is simply structured or sparse and if SNR is sufficient in the projection images. With this technique, both high temporal and spatial resolution can be achieved simultaneously. PMID- 16270331 TI - Combination of optimized transmit arrays and some receive array reconstruction methods can yield homogeneous images at very high frequencies. AB - Image inhomogeneity related to high radiofrequencies is one of the major challenges for high field imaging. This inhomogeneity can be thought of as having 2 radiofrequency-field related contributors: the transmit field distribution and the reception field distribution. Adjusting magnitude and phase of currents in elements of a transmit array can significantly improve flip angle homogeneity at high field. Effective application of some well-known parallel imaging and other receive array post-processing methods removes receptivity patterns from the intensity distribution in the final image, though noise then becomes a function of position in the final image. Here simulations are used to show that, assuming high signal-to-noise ratio, very homogeneous images in the human head can be acquired with the combination of transmit arrays and some receive array reconstruction methods at frequencies as high as 600 MHz. PMID- 16270332 TI - Fully automated shim mapping method for spectroscopic imaging of the mouse brain at 9.4 T. AB - For spectroscopic imaging studies of the mouse brain, it is critical to obtain optimal B(0) homogeneity over a large region of interest (ROI). In this paper, a fully automated shimming method for mouse brain at 9.4 T, based on B(0) mapping, is described. B(0) maps were obtained using a multislice gradient echo sequence with multiple phase evolution time delays with a novel unwrapping scheme. The unwrapping method allows phase maps with large bandwidths (+/-1 kHz) but with high resolution (0.3 Hz/ degrees ) to be acquired in a single acquisition, thereby minimizing the number of iterations required. The SD of the B(0) over the ROI (8 x 5 x 1 mm) was less than 10 Hz after shimming. Application of this method to the in vivo mouse brain allowed reproducible, high-quality spectroscopic data to be collected with 1-microl voxels. PMID- 16270333 TI - B(1) destructive interferences and spatial phase patterns at 7 T with a head transceiver array coil. AB - RF behavior in the human head becomes complex at ultrahigh magnetic fields. A bright center and a weak periphery are observed in images obtained with volume coils, while surface coils provide strong signal in the periphery. Intensity patterns reported with volume coils are often loosely referred to as "dielectric resonances," while modeling studies ascribe them to superposition of traveling waves greatly dampened in lossy brain tissues, raising questions regarding the usage of this term. Here we address this question experimentally, taking full advantage of a transceiver coil array that was used in volume transmit mode, multiple receiver mode, or single transmit surface coil mode. We demonstrate with an appropriately conductive sphere phantom that destructive interferences are responsible for a weak B(1) in the periphery, without a significant standing wave pattern. The relative spatial phase of receive and transmit B(1) proved remarkably similar for the different coil elements, although with opposite rotational direction. Additional simulation data closely matched our phantom results. In the human brain the phase patterns were more complex but still exhibited similarities between coil elements. Our results suggest that measuring spatial B(1) phase could help, within an MR session, to perform RF shimming in order to obtain more homogeneous B(1) in user-defined areas of the brain. PMID- 16270334 TI - Physiology and pathology of notch signalling system. AB - Notch proteins encode a family of transmembrane receptors that are part of a signalling transduction system known as Notch signalling, an extremely conserved and widely used mechanism regulating programs governing growth, apoptosis and differentiation in metazoans. Notch signalling begins when the Notch receptor binds ligands and ends when the Notch intracellular domain enters the nucleus and activates transcription of target genes. This core pathway is subjected to a wide array of regulatory influences and protein-protein interactions and is correlated with other signalling pathway. This review will summarize recent findings concerning the physiology and pathology of Notch signalling in vascular development and homeostasis. Moreover, the clinical phenotypes of Notch3 signalling system pathology will be described, with particular regard to CADASIL (Cerebral Autosomal Dominant Arteriopathy with Subcortical Infarcts and Leukoencephalopathy) for which the most recent pathogenetic hypotheses are reported. PMID- 16270335 TI - Bone response to 3D periodic hydroxyapatite scaffolds with and without tailored microporosity to deliver bone morphogenetic protein 2. AB - Three types of model hydroxyapatite (HA) scaffolds were implanted in the metacarpal and metatarsal bones of goats. Scaffolds, consisting of a latticed pattern of rods, were fabricated with a solid freeform fabrication (SFF) technique. All scaffolds contained macropores; some were also fabricated with micropores (5.2 +/- 2.0 microm). Recombinant human bone morphogenetic protein-2 (rhBMP-2) was added to some microporous scaffolds. rhBMP-2 caused increased percent filled with bone tissue compared to microporous scaffolds without rhBMP 2. Lamellar bone in the scaffolds was aligned perpendicular to the long axis of the bone near the junctions of the rods that make up the scaffold but was more random away from the junctions of rods. Microporous scaffolds stained beneath areas of contact with new bone. This staining might indicate either extracellular matrix (ECM) in the rods, byproducts of ECM production, or reaction of cellular products with the scaffold. PMID- 16270336 TI - Determining the degree of cortical bone asymmetry in bilateral, nonpathological, human femur pairs. AB - When testing the effects of a femoral component on cortical bone following total hip arthroplasty, the patient's implanted femur is often compared with his/her contralateral nonimplanted femur, with differences attributed to the femoral component. However, if normal anatomical differences exist between bilateral femurs, they need to be quantified in order to validate whether the differences between implanted and nonimplanted bilateral femurs are due to the implant or possibility due to intrinsic differences before implantation. This study quantified the geometric properties of cortical bone shape between seven pairs of bilateral, cadaveric, human femurs. The null hypothesis tested stated that the bilateral femurs would not be significantly different in cortical bone geometry. Digitized images of cortical bone cross-sections taken at percent biomechanical lengths (levels 1-8) were used to calculate bone geometry measurements. The paired t-test showed that the only significant difference was in the location of principal axes at the most proximal location, level 1 (p = 0.015). All other measurements and levels were not significant with percent differences less than 6.6%. In conclusion, the data supports attributing cortical bone shape differences between implanted and contralateral nonimplanted femurs in levels 2-8 to the presence of the implant when the significant differences are greater than 6.6%. PMID- 16270337 TI - Fabrication and evaluation of porous 2,3-dialdehydecellulose membrane as a potential biodegradable tissue-engineering scaffold. AB - A simple, novel method to produce porous 2,3-dialdehydecellulose (DAC) membranes as a potential tissue-engineering scaffold has been developed from methylolcellulose by the simultaneous water-induced phase separation and sodium chloride salt leaching techniques, followed by oxidation with sodium periodate in water. Membrane pores increased in size with increasing weight or particle size of the sodium chloride salt. The porosity of the membrane was not affected by the salt particle size, but it increased with an increase in the salt weight to 60%. At higher salt weight percentages, no significant change in the membrane porosity was observed. The oxidation step had no effect on the membrane porosity or pore size. All membranes with a porosity value ranging between 87 and 93% showed interconnected porous structures. The use of these membranes as a potential tissue-engineering scaffold was evaluated with the use of human neonatal skin fibroblast cells. Confocal microscopy showed cell attachment and spreading on these membranes. Immunohistochemical tests revealed the presence of collagen type III and fibronectin, indicating that the cells were viable and formed the extracellular matrix. In conclusion, the DAC membrane supports cell adhesion and proliferation and hence shows potential to be used as a tissue-engineering scaffold. PMID- 16270338 TI - Plasma-immersion ion-implanted nitinol surface with depressed nickel concentration for implants in blood. AB - Ion implantation into nitinol had been shown previously to decrease the surface nickel concentration of this alloy and produce a titanium oxide layer. Nothing is known yet about the blood compatibility of this surface and the suitability for implants in the blood vessels, like vascular stents. Nickel depletion of superelastic nitinol was obtained by oxygen or helium plasma-immersion ion implantation. The latter leads to the formation of a nickel-poor titanium-oxide surface with a nanoporous structure, which was used for comparison. Fibrinogen adsorption and conformation changes, blood platelet adhesion, and contact activation of the blood clotting cascade have been checked as in vitro parameters of blood compatibility; metabolic activity and release of cytokines IL-6 and IL-8 from cultured endothelial cells on these surfaces give information about the reaction of the blood vessel wall. The oxygen-ion-implanted nitinol surface adsorbed less fibrinogen on its surface and activated the contact system less than the untreated nitinol surface, but conformation changes of fibrinogen were higher on the oxygen-implanted nitinol. No difference between initial and oxygen implanted nitinol was found for the platelet adherence, endothelial cell activity, or cytokine release. The nanoporous, helium-implanted nitinol behaved worse than the initial one in most aspects. Oxygen-ion implantation is seen as a useful method to decrease the nickel concentration in the surface of nitinol for cardiovascular applications. PMID- 16270339 TI - Attachment, growth, and activity of rat osteoblasts on polylactide membranes treated with various low-temperature radiofrequency plasmas. AB - Nonporous and porous membranes from poly(L/DL-lactide) 80/20% were treated with low-temperature oxygen, ammonia, or sulphur dioxide-hydrogen plasmas and the late effects of plasma treatment on physicochemical characteristics of the membranes' surface were analyzed. The plasma treatment resulted in the permanent attachment of sulphur and nitrogen functionalities to the membrane's surface, and increased the surface concentration of oxygen, thereby increasing the surface wettability. To assess whether the plasma treatment affects the cellular response, primary rat osteoblasts were cultured on nontreated and plasma-treated nonporous and microporous membranes, and attachment, growth, and activity of cells were investigated. It was found that attachment and growth of osteoblasts on all the plasma-treated membranes were greater compared with nontreated controls. The treatment with ammonia plasma was most efficacious. The beneficial effects of plasma treatment on cells were most pronounced for microporous polylactide membranes irrespective of the plasma used. The results of the study suggest that the treatment of porous polylactide structures with plasma can be an effective means of enhancing their suitability for tissue engineering. Plasma exposure may also have an advantageous effect on bone healing when polylactide membranes are used to treat bone defects. PMID- 16270340 TI - Thin calcium phosphate coatings on titanium by electrochemical deposition in modified simulated body fluid. AB - Adherent and optically semitransparent thin calcium phosphate (CaP) films were electrochemically deposited on titanium substrates in a modified simulated body fluid at 37 degrees C. Coatings deposited by using periodic pulsed potentials showed better adhesion and better mechanical properties than coatings deposited with use of a constant potential. Scanning electron microscopy was used to study the morphology of the coatings. The coatings displayed a polydispersed porous structure with pores in the range of a few nanometers to 1 mum. Furthermore, X ray diffractometry and the O(1s) satellite peaks in X-ray photoelectron spectroscopy indicated that the coatings possessed a similar surface chemistry to that of natural bone minerals. These results were confirmed by inductively coupled plasma optical emission spectrometry, which yielded a Ca:P ratio of 1.65, close to that of hydroxyapatite. Contact mode atomic force microscopy (AFM) showed the average thickness of the coatings was in the order of 200 nm. Root mean-square (RMS) roughness values, also derived by AFM, were shown to be much higher on the titanium-CaP surfaces in comparison with untreated titanium substrates, with RMS values of about 300 and 110 nm, respectively. Cell culture experiments showed that the CaP surfaces are nontoxic to MG63 osteoblastic cells in vitro and were able to support cell growth for up to 4 days, outperforming the untreated titanium surface in a direct comparison. These easily prepared coatings show promise for hard-tissue biomaterials. PMID- 16270341 TI - Extensive H(+) release by bone substitutes affects biocompatibility in vitro testing. AB - Bone substitutes are widespread in orthopedic and trauma surgery to restore critical bony defects and/or promote local bone healing. Cell culture systems have been used for many years to screen biomaterials for their toxicity and biocompatibility. This study applies a human bone marrow cell culture system to evaluate the toxic in vitro effects of soluble components of different bone substitutes, which are already in clinical use. Different specimens of tricalcium phosphates (TCP) (Vitoss, Cerasorb), nondecalcified bovine bone (Lubboc), demineralized human bone matrices (DBM) (Grafton Flex/Putty), and collagen I/III matrix (ACI-Maix) were tested in Dulbecco's modified Eagle's medium (DMEM) and MesenCult culture solution and compared with a biomaterial-free cell culture. Biocompatibility parameters were cell viability evaluated by phase-contrast microscopy and laser flow cytometry, morphology, and the local H(+) release by bone substitutes. There were significant differences (p < 0.05) between the tested biomaterials and culture solutions. Collagen I/III, non-demineralized bovine bone, and TCP materials showed advantages for cell survival over other tested biomaterials (average values of vital cells/mL MesenCult/DMEM: Collagen I/III: 1090/1083; Vitoss: 893/483; Cerasorb: 471/523; Lubboc: 815/410; Grafton Putty: 61/44; Grafton Flex: 149/57). Especially the DBM materials lead to a significant decrease of pH, which is considered to be a major factor for cell death. DMEM culture solution supports cell survival for those bone substitutes that induce an alkaline reaction, whereas MesenCult media promotes cell vitality in biomaterials, which leads to an acidification of culture solution. PMID- 16270342 TI - Cancer and the family: an integrative model. AB - The stresses and uncertainties of cancer can be overwhelming for families. The Family System Illness model offers a psychosocial map to address these challenges and to make the inevitable strains more manageable. It provides a set of terms and a common language within a framework that attends to the longitudinal, psychosocial pattern of a health condition within a multigenerational, life cycle, and belief system context. Such a psychosocial framework can facilitate collaborative, effective coping and adaptation, thereby enhancing the quality of life for families affected by cancer. The model's utility is discussed for research, preventive screening, family assessment, treatment planning, and service delivery in a wide range of healthcare settings. PMID- 16270344 TI - Enhancing surface free energy and hydrophilicity through chemical modification of microstructured titanium implant surfaces. AB - Roughness-induced hydrophobicity, well-known from natural plant surfaces and intensively studied toward superhydrophobic surfaces, has currently been identified on microstructured titanium implant surfaces. Studies indicate that microstructuring by sandblasting and acid etching (SLA) enhances the osteogenic properties of titanium. The undesired initial hydrophobicity, however, presumably decelerates primary interactions with the aqueous biosystem. To improve the initial wettability and to retain SLA microstructure, a novel surface modification was tested. This modification differs from SLA by its preparation after acid etching, which was done under protective gas conditions following liquid instead of dry storage. We hypothesized that this modification should have increased wettability due to the prevention of contaminations that occurs during air contact. The main outcome of dynamic wettability measurements was that the novel modification shows increased surface free energy (SFE) and increased hydrophilicity with initial water contact angles of 0 degrees compared to 139.9 degrees for SLA. This hydrophilization was kept even after any drying. Reduced hydrocarbon contaminations were identified to play a possible role in altered surface thermodynamics. Such surfaces aim to retain the hydrophilicity and natural high surface energy of the Ti dioxide surface until surgical implants' insertion and are compared in this in vitro study with structural surface variants of titanium to compare roughness and chemically induced wettability. PMID- 16270343 TI - Effect of bioactive ceramic dissolution on the mechanism of bone mineralization and guided tissue growth in vitro. AB - A major objective of this research work was to evaluate the effect of bone cells on the dissolution-precipitation reaction in vitro. Rat bone marrow stem cells were seeded on silica-calcium phosphate nano composite (SCPC) with different chemical compositions and crystalline structures. Measurements of the Ca, P, Si, and Na concentrations in the tissue culture media using inductively coupled plasma indicated that bone marrow stem cells attached to the surface of SCPC did not affect the dissolution behavior of the material. However, bone marrow stem cells interfered with the back precipitation reaction and inhibited the formation of a calcium phosphate (Ca-P) layer on the material surface. Scanning electron microscope-energy-dispersive X-ray analyses showed that, in the absence of cells, a Ca-P layer formed on the material surface because of the dissolution precipitation reaction. Bone cells attached to SCPC that contains high silica content absorbed significantly higher concentrations of medium Ca than cells attached to SCPC that contains low silica content. In conjunction with the absorption of high Ca concentration, attached bone marrow stem cells produced calcified nodules and mineralized extracellular matrix, indicating osteoblastic differentiation. Results of the study strongly suggest that the mechanism of bone mineralization at the interface with bioactive ceramics is mainly cell mediated and is enhanced by the absorption of critical concentrations of dissolved Ca and P. The silicon-rich phase also provided a guided cell adhesion and tissue growth in vitro. The enhanced bioactivity reactions and strong stimulatory effect on bone cell function are attributed to the modified crystalline structure of the SCPC material. PMID- 16270345 TI - Three-dimensional culture of human osteoblastic cells in chitosan sponges: the effect of the degree of acetylation. AB - In this investigation, the effect of the degree of acetylation (DA) of chitosan on the behavior of human osteoblastic MG-63 cells cultured in three-dimensional chitosan matrices was assessed. Chitosan sponges with DAs in the range of 4 to 49% were prepared and characterized in terms of microstructure, porosity, and pore size. Collagen sponges were used as 3D control. Cell proliferation was determined using the MTT assay while the retention of the osteoblastic phenotype was monitored by assaying alkaline phosphatase activity. Cell morphology, cytoskeletal organization, and viability were assessed using different microscopy techniques. Chitosan sponges showed a similar microstructure regardless the DA, except for the highest DA used, where a more heterogeneous pore distribution was observed. In terms of cell proliferation, alkaline phosphatase activity and cell viability, cells cultured in chitosan scaffolds performed as well as in the 3D control regardless the DA, except for the highest DA used, where an inhibitory effect on cell proliferation was found. However, while in sponges with DAs < or = 13% cells attached and spread displaying long cell filopodia and numerous cell-to cell contacts, in sponges with higher DAs cells tended to remain spherical and grow into spheroid-like cellular aggregates. In the present study, the DA played a key role in determining the affinity of osteoblastic cells towards the substrates, possibly by influencing the nature of the initial adsorbed protein layer. PMID- 16270346 TI - In vitro enzymatic stability of dendritic peptides. AB - PEGylated polyamidoamine (PAMAM) dendrimers as drug carriers have been a topic of interest because of their biomedically favorable features, including minimal toxicity, reduced immunogenicity, and excellent solubility in aqueous and most organic solutions. A PEG shell on dendrimer surface may provide steric hindrance, known as stealth properties of PEG, to stabilize drug molecules to be delivered. In this article, the effects of PEG and coupling sequence of drug, PEG, and dendrimer in modulating the stability of delivered drug molecules were evaluated. N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide was chosen as a model peptide. Dendritic peptides, that is, peptide-dendrimer, peptide-PAMAM-PEG, and peptide PEG-dendrimer, were constructed based on Starbursttrade mark G3.0 PAMAM dendrimer and characterized by (1)H-NMR spectroscopy. Hydrolysis of dendritic peptides was catalyzed by alpha-chymotrypsin in pH 7.4 PBS buffer containing 5% DMF (v/v) at room temperature. The enzymatic stability of dendritic peptides was peptide-PAMAM PEG > peptide-PAMAM > free peptide > peptide-PEG-PAMAM. The ratio of PEG/peptide could be reduced for increasing peptide loading while maintaining the delivered peptides' relatively high enzymatic stability. The quantitative analysis of dendritic peptide/enzyme interactions provided the understandings of the molecular structure/stability relationships of dendrimer/drug for the design of an optimal PEGylated dendrimer-based drug-delivery system. PMID- 16270347 TI - A long-term in vitro biocompatibility study of a biodegradable polyurethane and its degradation products. AB - The biological safety of degradation products from degradable biomaterials is very important. In this study a new method is proposed to test the cytotoxicity of these degradation products with the aim to save time, laboratory animals, and research funds. A biodegradable polyurethane (PU) foam was subjected to this test method. The PU had soft segments of DL-lactide/epsilon-caprolactone and hard segments synthesized from butanediol and 1,4-butanediiosocyanate. Copolymer foams without urethane segments, consisting of DL-lactide/epsilon-caprolactone, were tested as well. Accumulated degradation products were collected by degrading the foams in distilled water at 60 degrees C up to 52 weeks. Cell-culture medium was prepared from powder medium with this water. In different tests the cytotoxicity of this medium was established. The first signs of cytotoxicity were observed after 3-5 weeks of degradation. This accounts for both materials and reestablishes the good short-term biocompatibility of these materials. The PU showed more toxicity toward the end stages of degradation in comparison with the copolymer. This is probably related to the accumulation of degradation products of the urethane segments. In the in vivo situation the degradation of the PU and the metabolism and excretion of degradation products may differ. Therefore, long term in vivo studies will have to establish whether these in vitro results are representative for the in vivo behavior of the degrading PU. PMID- 16270348 TI - Guided tissue regeneration for using a chitosan membrane: an experimental study in rats. AB - Barrier membranes are employed clinically to deflect the growth of gingival tissues away from root surface. They provide an isolated space over the regions with the defective tissues that allow the relatively slow growing periodontal ligament fibroblasts to be repopulated onto the root surface. Several makes of bioabsorbable membranes are now commercially available. In this study, we have employed chitosan as barrier membrane material and evaluated it for a guided tissue regeneration application. Three types of chitosan membranes: Chi-NaOH, Chi Na(5)P(3)O(10), and Chi-Na(2)SO(3)(each was gelated by NaOH, crosslinked by Na(5)P(3)O(10) and Na(2)SO(3), respectively), were prepared to be evaluated by the following categories: the mechanical strength to create an effective space, the rapid rate to reach hydrolytic equilibrium in phosphate-buffered solution, and the ease of clinical manipulative operations. Consequently, standardized, transosseous and critical sized skull defects were made in adult rats and the defective regions were covered with the specifically prepared chitosan membranes. After 4 weeks of recovering, varying degrees of bone healing were observed beneath the chitosan membranes in comparison to the control group. The chitosan covered regions showed a clear boundary space between connective tissues and bony tissues. Apparently, this process resulted in a good cell occlusion and beneficial osteogenesis effect to the bone. As for the control group, the bone defect was filled with connective tissue, and a destruction of the integrity of newly formed bone was observed. Among the chitosan membranes tested in this study, Chi-NaOH membrane provided a higher percentage of new bone formation than those from the Chi-Na(5)P(3)O(10) and Chi-Na(2)SO(3) families. PMID- 16270349 TI - New physically and chemically crosslinked hyaluronate (HA)-based hydrogels for cartilage repair. AB - When dissolved in aqueous solutions, sodium hyaluronate substituted with low amounts of alkyl chains [amphiphilic hyaluronate (HA)] can give rise to hydrogels thanks to intermolecular reversible hydrophobic interactions, leading to a three dimensional (3D) network. Such hydrogels possess shear-thinning properties and can thus be injected in cartilage defect to promote chondrocyte proliferation and cartilage repair. However, these hydrogels are only physically crosslinked and can progressively loose their 3D structure when they are in contact with aqueous fluids. To overcome this drawback, HA derivatives substituted with dodecyl chains were chemically crosslinked by a difunctional reagent, tetraethylene glycol ditosylate (TEG-diOTs). To preserve the shear-thinning properties of amphiphilic HA, small amounts of TEG-diOTs were used so as to obtain a low chemical crosslinking ratio. After optimization of the synthesis parameters, aqueous solutions of the HA derivatives, crosslinked both physically and chemically, were obtained, with rheological properties improved compared to the amphiphilic polymers. As the hydrogels are aimed to cartilage repair, they were sterilized by wet heating; the effect of this treatment on the polymer characteristics was analyzed by different techniques. A similar study was carried out on HA derivatives stored under conditions mimicking physiological ones. PMID- 16270350 TI - Detection of biofilm formation in Staphylococcus epidermidis from implant infections. Comparison of a PCR-method that recognizes the presence of ica genes with two classic phenotypic methods. AB - Biofilm-forming ability is increasingly being recognized as an important virulence factor in Staphylococcus epidermidis. This study compares three different techniques for the detection of biofilm-positive strains. The presence of icaA and icaD genes responsible for biofilm synthesis was investigated by a PCR method in a collection of 80 S. epidermidis strains isolated from orthopedic implant infections. The results from molecular analysis were compared with those obtained by two classic phenotypic methods, the Congo red agar (CRA) plate test and the microtiter plate test (MtP). Fifty-seven percent of all the examined strains were found icaA/icaD-positive, of which only three were not positive for CRA test. Differently, by the MtP method, 66% of the strains were found to be biofilm-producers but only a limited agreement with the PCR-method was noticeable because of the observation of (icaA/icaD+)/MtP- strains (8%) and of a surprising ambiguous result of (icaA/icaD-)/MtP+ strains (16%). The category of the weak biofilm-producers provided the highest contribution to these mismatching results (10%). The better agreement between the CRA plate test with the molecular detection of ica genes indicates the former as a reliable test for the phenotypic characterization of virulence of clinical isolates. However, MtP method remains a precious tool for the in vitro screening of different biomaterials for the adhesive properties using a reference strain. PMID- 16270351 TI - Scar myofibroblasts of the infarcted rat heart express natriuretic peptides. AB - The present study examined whether natriuretic peptide expression in the scar of post-myocardial infarcted (MI) rats was derived at least in part by residing myofibroblasts. ANP and BNP mRNA levels were significantly increased in the non infarcted left ventricle and scar of 1-week post-MI male rats, as compared to the left ventricle of normal rats. The infarct region contained myofibroblasts and contracted cardiac myocytes residing predominantly in the epicardial border zone. In primary passage scar-derived myofibroblasts, alpha-myosin heavy chain mRNA was undetectable, whereas ANP, BNP, as well as adrenomedullin and corin mRNA expression persisted. In 1-3 day cultured primary passage myofibroblasts, prepro ANP, mature ANP, and BNP staining was observed in the cytoplasm/perinuclear region co-incident with unorganized alpha-smooth muscle actin. Following 4-7 days in culture, myofibroblasts expressed organized alpha-smooth muscle actin filaments. However, natriuretic peptides were predominantly detected in the nucleus and cytoplasm, and thin filaments occupying the perinuclear region were positive for prepro-ANP and BNP. Isoproterenol treatment of first passage scar myofibroblasts increased protein synthesis and induced BNP mRNA expression, whereas ANP mRNA levels remained unchanged. By contrast, neither ANP nor BNP mRNAs were induced following exposure to AII despite increased protein synthesis. These data highlight the novel observation that scar myofibroblasts synthesized ANP, BNP, adrenomedullin, and expressed the pro-convertase corin. Constitutive and sympathetic-driven natriuretic peptide synthesis by myofibroblasts may in part influence reparative fibrosis. PMID- 16270352 TI - Platelet-derived growth factor-BB-induced hypertrophy of peritubular smooth muscle cells is mediated by activation of p38 MAP-kinase and of Rho-kinase. AB - Peritubular smooth muscle cells (PSMC) from rat testis in primary serum-free cultures unexpectedly undergo contraction and subsequent cell hypertrophy in response to the growth factor PDGF-BB, remaining stationary. The present study investigates the transduction pathways involved in the observed paradoxical upregulation of the differentiated phenotype and induction of hypertrophy in PSMC. PI3K, ERK, JNK, and p38 kinases, known to mediate PDGF-BB signaling in the canonic dedifferentiative and proliferative response of smooth muscle cells (SMC) were rapidly activated by PDGF-BB but only p38 remained activated after 2-day stimulation. Immunofluorescence and immunoblotting experiments showed that in 4 day treatment: (i) continuous inhibition of PI3K, of ERK, of JNK, failed to inhibit either cell enlargement and formation of prominent alpha-SM actin containing stress fibers or the typical increase in alpha-SM actin; (ii) when stimulated in the presence of the p38 inhibitor SB203580 both responses were significantly inhibited and cytofluorimetric analysis of cell size showed a remarkable reduction of the hypertrophic response. PDGF-BB was also found to activate the small GTPase RhoA and inhibition of Rho-dependent kinase ROCK by Y27632 counteracted the effects of PDGF-BB similarly to SB203580. Both the transcription factor ATF2 and the nucleosomal kinase MSK1, downstream targets of p38, were activated by PDGF-BB, but p38 inhibitor SB203580 inhibited only the phosphorylation of MSK1 which appeared unaffected by ROCK inhibitor Y27632. In concluding, p38 and the Rho-ROCK system were found to play prominent, probably independent roles in the upregulation of PSMC differentiated phenotype and induction of hypertrophy by PDGF-BB. PMID- 16270353 TI - Four novel RUNX2 mutations including a splice donor site result in the cleidocranial dysplasia phenotype. AB - Cleidocranial dysplasia (CCD) is an autosomal dominant disorder caused by haploinsufficiency of the RUNX2 gene. In this study, we analyzed by direct sequencing RUNX2 mutations from eleven CCD patients. Four of seven mutations were novel: two nonsense mutations resulted in a translational stop at codon 50 (Q50X) and 112 (E112X); a missense mutation converted arginine to glycine at codon 131 (R131G); and an exon 1 splice donor site mutation (donor splice site GT/AT, IVS1 + 1G > A) at exon 1-intron junction resulted in the deletion of QA stretch contained in exon 1 of RUNX2. We focused on the functional analysis of the IVS1 + 1G > A mutation. A full-length cDNA of this mutation was cloned (RUNX2Deltae1) and expressed in Chinese hamster ovary (CHO) and HeLa cells. Functional analysis of RUNX2Deltae1 was performed with respect to protein stability, nuclear localization, DNA binding, and transactivation activity of a downstream RUNX2 target gene. Protein stability of RUNX2Deltae1 is similar to wild-type RUNX2 as determined by Western blot analysis. Subcellular localization of RUNX2Deltae1, assessed by in situ immunofluorescent staining, was observed with partial retention in both the nucleus and cytoplasm. This finding is in contrast to RUNX2 wild-type, which is detected exclusively in the nucleus. DNA binding activity was also compromised by the RUNX2Deltae1 in gel shift assay. Finally, RUNX2Deltae1 blocked transactivation of the osteocalcin gene determined by transient transfection assay. Our findings demonstrate for the first time that the CCD phenotype can be caused by a splice site mutation, which results in the deletion of N-terminus amino acids containing the QA stretch in RUNX2 that contains a previously unidentified second nuclear localization signal (NLS). We postulate that the QA sequence unique to RUNX2 contributes to a competent structure of RUNX2 that is required for nuclear localization, DNA binding, and transactivation function. PMID- 16270354 TI - Role of the RANKL/RANK system in the induction of interleukin-8 (IL-8) in B chronic lymphocytic leukemia (B-CLL) cells. AB - B chronic lymphocytic leukemia (B-CLL) cells express several members of the tumor necrosis factor (TNF) family, such as CD40L, CD30L, and TRAIL. By using the cDNA microarray technology, B-CLL samples were found to overexpress receptor activator of nuclear factor kB (NF-kB) ligand (RANKL), as compared to normal CD19(+) B cells. These findings were validated at the protein level by Western blot and flow cytometry analyses. Moreover, unlike primary normal B cells, leukemic B-CLL cells showed surface expression of RANK, the cognate transmembrane receptor of RANKL. When added in vitro to B-CLL cultures, either alone or in association with chlorambucil or fludarabine, recombinant RANKL did not significantly modulate cell viability, and it minimally affected the IL-8 expression/release. On the other hand, treatment with RANK-Fc chimera potently upregulated the release of IL 8 in the B-CLL culture supernatants, suggesting involvement of reverse signaling through transmembrane RANKL in IL-8 induction. In turn, exposure of B-CLL cells to recombinant IL-8 significantly decreased spontaneous apoptosis as well as chlorambucil- and fludarabine-mediated cytoxicity in B-CLL cells. Since IL-8 has been implicated in progression of B-CLL disease, our findings suggest that, by upregulating IL-8, the RANKL/RANK system may contribute to the pathogenesis of B CLL. PMID- 16270355 TI - Steering estrogen signals from the plasma membrane to the nucleus: two sides of the coin. AB - Estrogen mediate its biological effects through its association with estrogen receptors (ERs). They also regulate the expression of a variety of genes involved in distinct physiological processes, including development, metabolism, and reproduction. In addition, emerging data suggest that the estrogen-estrogen receptor complex can also function as a cytoplasmic signaling molecule and may influence processes such as cardiovascular protection, bone preservation, neuroprotection, and proliferation of various cell types. Such extranuclear or nongenomic signaling pathways are rapid and supposedly independent of transcription. A recent exciting finding was that G-coupled membrane protein receptor, GPR30, an alternative to the classical ERs, is also involved in the rapid signaling of estrogen through its direct association with estrogen. These new findings combined with the recent advances in the cytoplasmic functions of proline, glutamic acid, luecine rich protein 1 (PELP1), and metastatic tumor antigen 1 short form (MTA1s) have opened a new spectrum and raised several new concerns in the field of estrogen biology and put the attention to unveil many unknown mechanistic actions of estrogen in cellular physiology. In this review, we briefly summarize what is currently known of the cellular mechanisms and physiology of estrogen's nongenomic actions in various cellular systems used by ERs. PMID- 16270358 TI - Emergent stent graft isolation of a knife-related subclavian arterial venous fistula: lessons learned during long-term follow-up. AB - We present the unusual case of a 42-year-old gentleman who entered the trauma center with a hemothorax and intermittent bright red pulsatile blood from three stab wounds in the ipsilateral supraclavicular fossa. The patient was found on emergent angiography to have a large right subclavian artery to vein fistula that was treated successfully with a Dacron-covered stent. Angiography, 2 years following stent-graft isolation, confirmed a durable result with evidence of hemodynamically insignificant neointimal growth through the pores of the graft material. This case provides insight into the long-term effects of a Dacron covered stent in patients with traumatic arterial injury. We discussed the advantages and disadvantages of different graft materials based on these angiographic findings. PMID- 16270359 TI - Successful high-risk coronary angioplasty in a patient with cardiogenic shock under circulatory assist with a 16F axial flow pump. AB - We report the case of a 75-year-old patient who suffered from subacute myocardial infarction and severely impaired left ventricular ejection fraction (EF: 17%). Using a novel 16F left ventricular assist device we performed an angioplasty of the right coronary artery, and of the left anterior descending artery. As a result of the circulatory support the patient recovered from cardiogenic shock within 8 hr. At a pump speed of 45,000 rpm the axial flow pump generated flow rates up to 3.3 l/min. The 16F pump cannula was removed using local compression. The EF was 51% at 30-day follow-up examination. PMID- 16270360 TI - Possible anaphylactic reaction to taxus stent: a case report. AB - We report a patient who experienced an anaphylactic reaction during coronary artery stenting. Immediately after a polymer-based paclitaxel-eluting stent (Taxus, Boston-Scientific) is deployed, the patient developed profound hypotension and an erythematous rash. The case complicated with coronary spasm and a new thrombus formation making a second stent implantation necessary. The reaction was successfully reversed with epinephrine, steroid, antihistaminic, a 9 hr infusion of dopamine, and intraaortic balloon counterpulsation. PMID- 16270361 TI - Drug-eluting stenting is superior to bare metal stenting in saphenous vein grafts. AB - This study compared the outcomes of percutaneous coronary intervention (PCI) of saphenous vein grafts (SVGs) with drug-eluting stents (DES) with bare metal stents (BMS). PCI of degenerated SVG is associated with worse outcomes and high incidence of in-stent restenosis compared with PCI of native coronary arteries. There is a paucity of data on the outcomes of PCI of SVG with DES. Data from 223 consecutive patients who underwent PCI of SVG were imputed into a dedicated clinical database. We assessed the clinical outcomes at a mean follow-up of 9.1+/ 2.1 months. A total of 139 patients underwent PCI of SVG with DES and 84 patients with BMS. The mean age of the SVG was 7.6+/-3.8 years in the DES group and 7.7+/ 2.8 years in the BMS group (P=0.38). Procedural success was achieved in all patients except for one patient in the BMS group who underwent emergent coronary artery bypass graft surgery for SVG dissection. There were no other in-hospital cardiac events in both groups. There was one cardiac death in the DES group and three deaths in the BMS group (P=0.03). When compared to the BMS, PCI of SVG with DES was associated with a lower incidence of myocardial infarction (4.3% vs. 20.2%; P=0.04) and target vessel revascularization (10.1% vs. 36.9%; P=0.035). When compared with BMS, PCI of SVG with DES was associated with a lower incidence of death, myocardial infarction, and target vessel revascularization. PMID- 16270362 TI - Drug eluting stents for everyone and everything? PMID- 16270363 TI - Transcatheter techniques in the management of perioperative vascular obstruction. AB - The role of transcatheter intervention for the treatment of vascular obstruction is well documented in the preoperative or remote postoperative settings. More recently, the roles of angioplasty and stent implantation have been advocated as intraoperative and immediate postoperative strategies. As one considers the inherent advantages in this cooperative approach to congenital heart disease, the development of a truly hybrid interventional suite seems imperative. PMID- 16270364 TI - Solution NMR studies of an intrinsically unstructured protein within a dilute, 75 kDa eukaryotic protein assembly; probing the practical limits for efficiently assigning polypeptide backbone resonances. AB - This paper describes an efficient NMR strategy for assigning the backbone resonances of an intrinsically unstructured protein (IUP), p21-KID, bound to its biological target, Cdk2/cyclin A. In order to overcome the challenges associated with the high molecular weight (75 kDa) and low solubility of the ternary complex (0.2 mM), we used perdeuteration, TROSY, and high-sensitivity cryogenic NMR probes at high magnetic-field strengths (i.e. 16.4, 18.8 and 21.1 Tesla). p21-KID was also prepared by using specific amino acid isotope labels. Most importantly, we studied binary, subcomplexes that allowed resonance assignments to be made in stages. We show that subdomains of p21-KID folded within binary complexes into the same conformations as observed in the ternary, Cdk2/cyclin A complex. This is a general feature of IUPs, which often adopt highly extended conformations when bound to other proteins. This strategy is suitable for studies of IUPs within considerably larger biomolecular assemblies as long as the IUP can be uniformly and selectively isotope labeled. PMID- 16270365 TI - Tetrabutylammonium fluoride-mediated rapid alkylation reaction in microtiter plates for the discovery of enzyme inhibitors in situ. PMID- 16270366 TI - A DNA mimic made of non-nucleosidic phenanthrene building blocks. PMID- 16270367 TI - Intramolecular hydroamination of functionalized alkenes and alkynes with a homogenous zinc catalyst. PMID- 16270368 TI - From colloidal Co/CoO core/shell nanoparticles to arrays of metallic nanomagnets: surface modification and magnetic properties. AB - The magnetic properties of nanoparticles can be subject to strong variations as the chemical composition of the particle surface is modified. To study this interrelation of surface chemistry and magnetism, self-assembled layers of colloidal 9.5 nm Co/CoO core/shell nanoparticles were exposed to mild reactive hydrogen and oxygen plasmas. The consecutive oxygen/hydrogen plasma treatment transforms the particle layer into an array of metallic nanomagnets with complete reduction of the oxide and removal of the organic surfactants. The original arrangement of the particle array and the number of Co atoms per particle remains unchanged within the experimental error, and thus this is a possible route for the fabrication of ultrahigh-density magnetic bit structures from colloidal dispersions. The magnetic properties can be tuned by controlling the thickness of the surface oxide layer, which magnetically hardens the particles, as evidenced by element-specific magnetic hysteresis loops. PMID- 16270369 TI - Absolute configuration of C76 from optical rotatory dispersion. AB - The absolute configuration of C76 has been determined as (+)589-(fC)-C76 , for the first time, by comparing the experimental and predicted optical rotatory dispersion (ORD) patterns. The experimental ORD pattern was derived from the experimental electronic circular dichroism (ECD) spectrum using the Kramers Kronig (KK) transform. The theoretical ORD spectra were calculated in the resonant region using linear response theory, and also using the KK transform of the theoretical ECD spectrum, at different theoretical levels, namely BHLYP/6 31G*, B3LYP/6-31G*, BLYP/6-31G*, and HF/6-31G*. Good agreement noted between experimental and predicted spectra allows for an unambiguous determination of the absolute configuration. PMID- 16270371 TI - Synchrotron radiation circular dichroism spectroscopy of ribose and deoxyribose sugars, adenosine, AMP and dAMP nucleotides. AB - Synchrotron radiation circular dichroism (SRCD) spectra of ribose and deoxyribose sugars, adenosine, AMP and dAMP nucleotides and cyclic derivatives were measured in the vacuum ultraviolet region (down to 168 nm for sugars and 175 nm for adenine derivatives) and at different pH values (3, 6-7, 9-10) and temperatures (between 5 and 45 degrees C). The information content in the VUV region is important since the CD bands strongly depend on the chemical structure of the sugar, the presence and orientation of a phosphate group and the protonation state of adenine. On the other hand, single or double deprotonation of the phosphoric acid group has no influence on the spectra. We assign the vacuum ultraviolet (VUV) CD bands of the nucleoside and nucleotides to be due mainly to n-->pi* transitions in the adenine nucleobase based on a comparison with the absorption spectra. The CD bands of the sugars are due to n(O -->sigma*) transitions and are much smaller than the CD signal from the nucleotides in the VUV region. Bands are assigned to both pyranose and open-chain forms. PMID- 16270370 TI - Patterns with high rhythmicity levels in multicomponent Liesegang systems. AB - Liesegang banding is the display of rhythmic strata of precipitate as co precipitate ions interdiffuse in a gel medium. Complex periodic patterns as well as aperiodic structures could emerge, notably in systems where more than one salt is precipitated. The use of three cations (Co2+) Ni2+, and Mg2+) in the banded precipitation of their hydroxides resulted in an unusual pattern with a consistently increasing rhythmicity. A periodic structure marked by the succession of band multiplets with increasing number of bands (from singlets to doublets to triplets to quadruplets, consistently) was observed. Such rhythmic patterns are obtained as the initial Mg2+ concentration ([Mg2+]0), chosen as a control parameter, increases through a critical value. At high [Mg2+]0, the trend breaks after a long time elapses. Two types of bifurcation are therefore experienced by such a system: concentration bifurcation and diffusive (time/space) bifurcation. The dynamics is elucidated on the basis of an analysis of the bands in certain groups, and gel regions between these groups, as well as between group blocs (here, a bloc denotes a succession of multiplet groups, with repetitively the same number of bands). Finally, similarities between our system and naturally occurring rhythmic patterns are emphasized and discussed. PMID- 16270372 TI - Reversible sequential-binding probe receptor-ligand interactions in single cells. AB - With the reversible sequential (ReSeq) binding assay,we present a novel approach for the ultrasensitive profiling of receptor function in single living cells. This assay is based on the repetitive application of fluorescent ligands that have fast association-dissociation kinetics. We chose the nicotinic-acetylcholine receptor (nAChR) as a prototypical example and performed ReSeq equilibrium, kinetic, and competition-binding assays using fluorescent derivatives of the antagonist alpha-conotoxin GI (alpha-CnTx). Thereby, we determined the binding constants of unlabeled alpha-CnTx and d-tubocurarine. The high selectivity of alpha-CnTx for muscle-type nAChR made it possible to observe specific binding even in the presence of other nAChR subtypes. Imaging of individual nAChRs and ligand-binding cycles to single cells in microfluidic devices demonstrated the ultimate miniaturization and accuracy of ReSeq-binding assays even at low receptor-expression levels. We expect our approach to be of generic importance for functional screening of compounds or membrane receptors, and for the detailed characterization of rare primary cells. PMID- 16270374 TI - Can acetylenedithiolate act as a tetradentate bridging ligand? PMID- 16270373 TI - Coumarinylmethyl esters for ultrafast release of high concentrations of cyclic nucleotides upon one- and two-photon photolysis. PMID- 16270375 TI - (Pr4N)4[Ag3Fe2(ECN)12]--anionic network structures with mutual interpenetration. PMID- 16270376 TI - Pulse pressure and mortality in hypertensive type 2 diabetic patients. A cohort study. AB - HYPOTHESIS: Hypertension is a well-known cardiovascular risk factor in type 2 diabetic patients. It has been suggested that pulse pressure (PP) could be an independent cardiovascular risk factor in the general population, particularly in the elderly. An association between office PP and cardiovascular mortality has been previously reported in diabetic patients, while the relationship between ambulatory measurements of PP and all-cause mortality has not been assessed so far. AIM: To assess the relationship between ambulatory PP and all-cause mortality in diabetic patients with hypertension. METHODS: A cohort study was performed on a consecutive series of 435 diabetic outpatients. All patients underwent office blood pressure measurement (OBP) and 24-h ambulatory blood pressure monitoring (ABPM). Mortality was assessed through queries at the Registry Offices of the city of residence for each patient. Mean follow-up was 3.8 +/- 1.2 years. RESULTS: Fifty-eight patients (13.3%) died during the follow up. Mortality was significantly (p < 0.05) higher in patients in the highest quartile and lower in patients in the lowest quartile, when compared to the intermediate quartiles, both for office and ABPM-PP. In a multivariate analysis, after adjustment for numerous variables (including current hypoglycaemic, antihypertensive statin and aspirin treatment), mortality was increased by 3.1 and 5.3% for each incremental mmHg of office PP (p < 0.05) and ABPM-PP (p < 0.001) respectively. CONCLUSIONS: High PP, assessed through office measurement or ABPM, was associated with increased mortality in hypertensive type 2 diabetic patients. In our sample, PP assessed with ABPM is a better predictor of mortality than office PP. PMID- 16270377 TI - Perspective: chromosomal aneuploidy in leukemia--lessons from down syndrome. AB - Abnormal number of chromosomes, aneuploidy, is the most common abnormality in leukemia and cancer. However, the casual relationship between aneuploidy and cancer is unclear. Additional copies of chromosome 21 are frequently found in leukemic cells. Constitutional trisomy 21 that characterizes Down Syndrome is associated with markedly increased risk for childhood leukemia. In this perspective I review recent studies that suggest that constitutional trisomy 21 promotes leukemic transformation during fetal hematopoiesis. As most of childhood leukemias arise in-utero, these studies are of general relevance to sporadic childhood leukemias. PMID- 16270378 TI - Evaluation of endometrial receptivity during in-vitro fertilization using three dimensional power Doppler ultrasound. AB - OBJECTIVE: To compare sonographic endometrial characteristics in in-vitro fertilization (IVF) cycles between women who conceive and those who do not. METHODS: Thirty-five women undergoing IVF treatment participated in the study. Using three-dimensional (3D) power Doppler ultrasound, we assessed endometrial patterns, volume and vascularization, after follicle stimulating hormone (FSH) stimulation but before human chorionic gonadotropin (hCG) administration (referred to hereafter as 'after FSH stimulation') and again on the day of oocyte retrieval. RESULTS: The pregnancy rate was 37% (13/35). After FSH stimulation, 29 of the 35 women had a triple-line endometrial pattern, compared with five out of 35 on the day of oocyte retrieval. In those who had a triple-line pattern after FSH stimulation the pregnancy rate was 44.8% (13/29) and it was 0% (0/6) in those with a homogeneous pattern (chi-square test, P = 0.039). If a triple-line pattern was present on the day of oocyte retrieval the pregnancy rate was 80.0% (4/5), whereas if the pattern was homogeneous the pregnancy rate was 30.0% (9/30) (P = 0.032). There were no differences between those who conceived and those who did not in endometrial thickness, volume or vascularization on either day examined. Endometrial volume decreased significantly after hCG injection in women who conceived, but not in those who did not conceive. In both groups endometrial and subendometrial vascularization decreased after hCG injection, while the endometrial thickness remained unchanged. CONCLUSIONS: The existence of a homogeneous endometrial pattern after FSH stimulation seems to be a prognostic sign of an adverse outcome in IVF, while a triple-line pattern after FSH stimulation and a decrease in endometrial volume appear to be associated with conception. PMID- 16270379 TI - Surface enhanced laser desorptions ionization-time of flight-mass spectrometry analysis in complex food and biological systems. PMID- 16270380 TI - Significance and prognostic value of lysosomal enzyme activities measured in surgically operated adenocarcinomas of the gastroesophageal junction and squamous cell carcinomas of the lower third of esophagus. AB - AIM: To establish whether there are fundamental differences in the biochemistries of adenocarcinomas of the gastroesophageal junction (GEJ) and the squamous cell carcinomas of the lower third of the esophagus (LTE). METHODS: Between February 1, 1997 and February 1, 2000, we obtained tissue samples at the moment of resection from 54 patients for biochemical analysis. The full set of data could be comprehensively analyzed in 47 of 54 patients samples (81%). Of these, 29 were adenocarcinomas of the GEJ Siewert type I (n = 8), type II (n = 12), type III (n = 9), and 18 presented as squamous cell carcinomas of the LTE. We evaluated the mean values of 11-lysosomal enzyme and 1-cytosol protease activities of the tumorous and surrounding mucosae as well as their relative activities, measured as the ratio of activity in tumor and normal tissues from the same patient. These data were further analyzed to establish the correlation with tumor localization, TNM stage (lymph-node involvement), histological type (papillary, signet-ring cell, tubular), state of differentiation (good, moderate, poor), and survival (or=24 mo). RESULTS: In adenocarcinomas, the activity of alpha mannosidase (AMAN), cathepsin B (CB) and dipeptidyl-peptidase I (DPP I) increased significantly as compared to the normal gastric mucosa. In squamous cell carcinomas of the esophagus, we also found a significant difference in the activity of cathepsin L and tripeptidyl-peptidase I in addition to these three. There was a statistical correlation of AMAN, CB, and DPP I activity between the level of differentiation of adenocarcinomas of the GEJ and lymph node involvement, because tumors with no lymph node metastases histologically confirmed as well-differentiated, showed a significantly lower activity. The differences in CB and DPP I activity correlated well with the differences in survival rates, since the CB and DPP I values of those who died within 24 mo following surgical intervention were significantly higher than of those who survived for 2 years or more. CONCLUSION: Adenocarcinomas of the GEJ form a homogenous group from a tumor-biochemical aspect, and differ from the biochemical characteristics of squamous cell carcinomas of the LTE on many points. When adenocarcinomas of the GEJs are examined at the preoperative phase, the ratio of the performed AMAN, CB, and DPP I enzymatic activity of the tissue sample from the tumor and adjacent intact mucosa within 2 cm of the tumor may have a prognostic value even in the preoperative examination period, and may indicate that ranking of these patients into the neo-adjuvant treatment group should be considered. PMID- 16270381 TI - Polymorphism of CYPIA1 and GSTM1 genes associated with susceptibility of gastric cancer in Shandong Province of China. AB - AIM: To explore whether polymorphisms of the CYPIA1 and GSTM1 genes are associated with susceptibility of stomach cancer. METHODS: A total of 102 stomach cancer cases and 62 healthy persons were diagnosed by pathology in 1998-2000 in the Qilu Hospital of Shandong University. Gene polymorphisms were detected by the PCR using sequence-specific primers. Data analysis of the case-control study was carried out using the unconditional logistic method. RESULTS: After adjustment for age, sex, educational levels, and occupation, the risk factors for stomach cancer were shown to be smoking, Helicobacter pylori (H pylori), and presence of the CYPIM G/G and GSTM1 O/O genotypes. Interaction was observed between the combined genotypes of either CYPIA1 G/G and GSTM1 O/O or H pylori infection, or GSTM1 O/O and H pylori infection or smoking. CONCLUSION: Polymorphisms of the CYPIA1 and GSTM1 genes, H pylori infection and smoking are related to susceptibility to stomach cancer. PMID- 16270382 TI - Inhibitory effects of N-(4-hydrophenyl) retinamide on liver cancer and malignant melanoma cells. AB - AIM: To investigate the effect of N-(4-hydrophenyl) retinamide (4-HPR), the derivative of retinoic acid, on inhibition of migration, invasion, cell growth, and induction of apoptosis in hepatocellular carcinoma cells (HCCs) and malignant melanoma cells. METHODS: 4-HPR was chemically synthesized. Cellular migration and invasion were assayed by Borden chamber experiment. Cell growth was assayed by MTT chromometry. Apoptosis effect was measured using Hoechst 32258 staining and flow cytometry. Gene transfection was performed with lipofectamine. RESULTS: We observed that the migration of HCC and melanoma cells was significantly suppressed by 4-HPR and the migration cells were reduced to 58+/-5.03 (control 201+/-27.2, P<0.05, n = 4) in SMMC 7721-k3 HCC, and to 254+/-25.04 (control 302+/ 30.1, P<0.05, n = 4) in melanoma cells after 6-h incubation with 4-HPR. The invasion through reconstituted basement membrane was also significantly reduced by 4-HPR treatment to 11.2+/-3.3 in SMMC 7721-k3 HCC (control 27+/-13.1), and to 24.3+/-3.2 in melanoma cells (control 67.5+/-10.1, P<0.05, n = 3). Cell growth, especially in melanoma cells, was also significantly inhibited. Furthermore, 3 micromol/L of 4-HPR induced apoptosis in B16 melanoma cells (37.11+/-0.94%) more significantly than all-trans retinoic acid (P<0.05), but it failed to induce apoptosis in SMMC 7721-k3 HCC. The mechanism for 4-HPR-induced apoptosis was not clear, but we observed that 4-HPR could regulate p27(kip1), and overexpression of cerebroside sulfotransferase (CST) diminished the apoptosis induced by 4-HPR in melanoma cells. CONCLUSION: 4-HPR is a potent inhibitor of HCC migration and inducer of melanoma cell apoptosis. CST and p27(kip1) expression might be associated with 4-HPR-induced apoptosis. PMID- 16270383 TI - Low prevalence of germline hMSH6 mutations in colorectal cancer families from Spain. AB - AIM: To investigate the prevalence and penetrance of hMSH6 mutations in Spanish HNPCC families that was negative for mutation in hMLH1 or hMSH2. METHODS: We used PCR-based DGGE assay and direct sequencing to screen for hMSH6 gene in 91 HNPCC families. RESULTS: we have identified 10 families with germ-line mutations in the DNA sequence. These mutations included two intronic variation, three missense mutation, one nonsense mutation, and four silent mutations. Among the 10 germ line mutations identified in the Spanish cohort, 8 were novel, perhaps, suggesting different mutational spectra in the Spanish population. Detailed pedigrees were constructed for the three families with a possible pathogenic hMSH6 mutation. The two silent mutations H388H and L758L, detected in a person affected of colorectal cancer at age 29, produce loss of the wild-type allele in the tumor sample. Immunohistochemical analysis showed that expression of MSH6 protein was lost only in the tumors from the carriers of V878A and Q263X mutations. CONCLUSION: Altogether, our results indicate that disease-causing germ line mutations of hMSH6 are very less frequent in Spanish HNPCC families. PMID- 16270384 TI - Consumption of purple sweet potato leaves modulates human immune response: T lymphocyte functions, lytic activity of natural killer cell and antibody production. AB - AIM: To study the immunological effects of physiological doses of purple sweet potato leaves (PSPL). METHODS: The randomized crossover study (two periods, each lasting for 2 wk) involved 16 healthy non-smoking adults of normal weight. The 6 wk study consisted of a run-in (wk 1) PSPL diet (daily consumption of 200 g PSPL) or a control diet (low polyphenols, with the amount of carotenoids adjusted to the same level as that of PSPL) (wk 2-3), washout diet (wk 4), and switched diet (wk 5-6). Fasting blood was collected weekly in the morning. T-lymphocyte function was assessed via the proliferation and secretion of immunoreactive cytokines. Salivary IgA secretion and the specific cytotoxic activities of cytotoxic T lymphocytes and natural killer (NK) cells were determined. RESULTS: The plasma beta-carotene level increased with time in both groups, while the plasma polyphenol level decreased in the control group, and no significant difference was detected between the two groups. Although plasma polyphenol levels did not significantly increase in the PSPL group at the end of the study, they were significantly elevated in urine. PSPL consumption produced a significant increase in proliferation responsiveness of peripheral blood mononuclear cells (PBMC) and their secretion of immunoreactive IL-2 and IL-4. As well, lytic activity in NK cells was elevated in a time-dependent fashion. Salivary IgA secretion significantly decreased in control group after 2 wk, and returned to baseline following dietary switch to PSPL. CONCLUSION: Consumption of PSPL modulates various immune functions including increased proliferation responsiveness of PBMC, secretion of cytokines IL-2 and IL-4, and the lytic activity of NK cells. The responsible determinants of PSPL remain to be elucidated, as does the biological significance of the present observations. PMID- 16270385 TI - Effect of Astragalus complanatus flavonoid on anti-liver fibrosis in rats. AB - AIM: To observe the anti-liver fibrosis effect of Astragalus complanatus flavonoids (ACF) in rats. METHODS: The liver fibrosis model in rats was established by injecting interperitoneally 0.2 mL/100 g 0.5% dimethylnitrosamine, thrice a week. Meanwhile, the rats were administered ACF (30, 60, 120 mg/kg) or colchicine (0.1 mg/kg) once a day for 1 mo. Serum N-propeptide of type I procollagen (PINP) and type III procollagen (PIIINP) were measured using ELISA. Malondialdehyde (MDA) and superoxide dismutase (SOD) in hepatic tissue were evaluated. Matrix metal protease-1 (MMP-1) mRNA expression was assayed by RT-PCR and the protein expression of tissue inhibitor of metal protease-1 (TIMP-1) was analyzed by immunohistochemistry. RESULTS: In the ACF groups, SOD activity increased and MDA content decreased in comparison to the liver fibrosis model group. The serum PINP and PIIINP contents in ACF-2 and -3 group decreased compared to those in model group. In ACF-2 and -3 group, the expression of MMP-1 mRNA increased significantly and the protein expression of TIMP-1 decreased compared to that in model group. CONCLUSION: The antifibrotic mechanisms of ACF are associated with its influence on lipid peroxidation and collagen synthesis and degradation. PMID- 16270386 TI - Amelioration of experimental colitis by Astragalus membranaceus through anti oxidation and inhibition of adhesion molecule synthesis. AB - AIM: To investigate the protective effects of Astragalus membranaceus (Am) against hapten-induced colitis in male Sprague-Dawley rats as well as its underlying mechanism. METHODS: Experimental colitis was induced in rats by enema administration of 2,4-dinitrobenzene sulfonic acid (DNBS). Rats were either pretreated with Am extract (2 or 4 g/kg, p.o. once daily) starting from 10 d before DNBS enema, or received Am post-treatment (2 or 4 g/kg, p.o. twice daily) on the three consecutive days following DNBS administration. Colonic lesion area and histological damage were determined, while the activities of myeloperoxidase (MPO) and xanthine oxidase, as well as reduced glutathione (GSH) content were measured in the excised colonic tissues. Besides, protein expression of inducible nitrite oxide synthase (iNOS), intercellular adhesion molecule-1 (ICAM-1) and P selectin was also detected by Western blot analysis. RESULTS: Our findings had shown that both macroscopic lesion area and histological colonic damage induced by DNBS were significantly reduced by both Am pre- and post-treatments. These were accompanied by attenuation of the elevated colonic MPO activity and downregulation of the iNOS, P-selectin, and ICAM-1 protein expression. Besides, deprivation of colonic GSH level under colitis condition was also preserved. CONCLUSION: These results demonstrate that Am possesses both preventive and therapeutic potential in experimental colitis. The anti-inflammatory actions involve anti-oxidation along with inhibition of adhesion molecule synthesis in the colonic tissues. PMID- 16270388 TI - Separation of growth-stimulating peptides for Bifidobacterium from soybean conglycinin. AB - AIM: To isolate and identify the soybean conglycinin peptides that selectively stimulates the growth of bifidobacteria in vitro, and to investigate the effect of soybean conglycinin peptides on intestinal ecosystem in vivo. METHODS: Soybean conglycinin was purified from soybean seeds by gel filtration (Sepharose-CL-6B). These proteins were submitted to hydrolysis by pepsin. Several growth-stimulating peptides for bifidobacteria were isolated chromatographically from pepsin hydrolysis of soybean conglycinin and identified by means of matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS). Parallel to in vitro study, in vivo experiments with soybean conglycinin peptides were performed in mice. Ninety male KM mice were randomly assigned into five groups of 16 mice each, and each group was administered for 21d intragastrically with physiological saline (control), conglycinin, pepsin-treated conglycinin (PTC), the most active fraction which isolated from pepsin-treated conglycinin (P2-PTC) and HCl-full hydrolysis of conglycinin (HCl-FHC), respectively. Intestinal microflora were evaluated by standard microbiologic methods and biochemical assays of cecal content samples after treatment. RESULTS: The results showed that the peptides which were isolated from soybean conglycinin could stimulate the growth of bifidobacteria in vitro, and the molecular mass of purified peptides with MALDI-TOF-MS ranged from 693.32 to 1829.55. Compared with control group, in vivo experiments showed that P2-PTC group decreased cecal pH (7.08+/-0.08 vs 7.21+/-0.09, P<0.05) and enterococci counts (5.38+/-0.26 log10CFU/g vs 5.78+/-0.19 log10CFU/g, P<0.05), significantly increased sIgA level (172.08+/-35.40 ng/g vs 118.27+/-33.93 ng/g, P<0.01) and beta-galactosidase activity (1.28+/-0.23 U/g vs 1.82+/-0.58 U/g, P<0.05). CONCLUSION: The results have shown that conglycinin is good source for enzyme-mediated production of peptides which stimulate the growth of bifidobacteria. These peptides are inactive within the sequence of the parent protein but can be released during enzymatic hydrolysis, and in vivo experiments demonstrate that conglycinin peptides may be beneficial for improving gastrointestinal health. PMID- 16270387 TI - Protective effect of selenium-enriched Lactobacillus on CCl4-induced liver injury in mice and its possible mechanisms. AB - AIM: To study the protective effects and mechanisms of Se-enriched lactobacillus on liver injury caused by carbon tetrachloride (CCl4) in mice. METHODS: Seventy two ICR mice were randomly divided into four groups: normal group, CCl4-induced model group, low Se-enriched lactobacillus treatment group (L-Se group), and high Se-enriched lactobacillus treatment group (H-Se group). During a 3-wk experimental period, the common complete diet was orally provided daily for normal group and model group, and the mice in L-Se and H-Se groups were given a diet with 2 and 4 mg of organoselenium from Se-enriched lactobacillus per kg feed, respectively. From the 2nd wk of experiment, the model group, L-Se group, and H-Se group received abdominal cavity injection of olive oil solution containing 500 mL/L CCl4 (0.07 mL/100 g body mass) to induce liver injury, and the normal group was given olive oil on every other day for over 2 wk. In the first 2 wk post injection with CCl4, mice in each group were killed. The specimens of blood, liver tissue, and macrophages in abdominal cavity fluid were taken. Then the activities of the following liver tissue injury-associated enzymes including glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) as well as malondialdehyde (MDA) content were assayed. Changes of phagocytic rate and phagocytic index in macrophages were observed with Wright-Giemsa stain. Plasma TNF-alpha level was measured by radioimmunoassay. The level of intracellular free Ca2+ ([Ca2+]i) in hepatocytes was detected under a laser scanning confocal microscope. RESULTS: During the entire experimental period, the AST and ALT activities in liver were greatly enhanced by CCl4 and completely blunted by both low and high doses of Se-enriched lactobacillus. The Se-enriched lactobacillus protected liver homogenate GSH-Px and SOD activities were higher or significantly higher than those in model group and were close to those in normal group. CCl4 significantly increased MDA content in liver homogenates, while administration of Se-enriched lactobacillus prevented MDA elevation. Phagocytic rate and phagocytic index of macro-phages decreased after CCl4 treatment compared to those in normal control, but they were dramatically rescued by Se-enriched lactobacillus, showing a greatly higher phagocytic function compared to model group. CCl4 could significantly elevate plasma TNF-alpha and hepatocyte [Ca2+]i level, which were also obviously prevented by Se-enriched lactobacillus. CONCLUSION: Se-enriched lactobacillus can intervene in CCl4-induced liver injury in mice by enhancing macrophage function activity to keep normal and beneficial effects, elevating antioxidant-enzyme activities and reducing lipid peroxidation reaction, inhibiting excessive release of TNF-alpha, preventing the dramatic elevation of [Ca2+]i in hepatocytes. PMID- 16270389 TI - Mesenchymal hamartoma of liver: magnetic resonance imaging and histopathologic correlation. AB - AIM: To describe the imaging features of hepatic mesenchymal hamartoma with emphasis on magnetic resonance imaging (MRI) compared to histopathologic results. METHODS: Spin-echo sequence(SE),fast spin-echo sequence(FSE) were performed in 12 children (7 males,5 females) with mesenchymal hamartoma of the liver (MHL), aged 1.2 months to 12 years;( mean age, 6.3 years) axial, saggital, coronary plain imaging with an Elscint 2.0T MR equipment. Their main symptoms were abdominal mass (5 cases), enlarged liver (8 cases), abdominal pain (1 case) and anemia (2 cases), and negative alphafetoprotein. Dynamic enhancement examination was added in 2 cases. RESULTS: Six cases had single mass type of MHL, in which 3 cases had solid masses showing slight low-signal-intensity in T1WI, and irregular high signal-intensity in T2WI, 1 case had a cystic-solid mixed mass showing several border-clear cysts in a solid mass, 2 cases had cystic masses with multi-septa. Five cases had diffuse and multifocal lesions type of MHL with its signal intensity being similar to that of the solid mass. One case had a combined diffuse and single cystic mass. In the early dynamic enhancement examination, the lesions were slightly circum-enhanced , and the center was enhanced in the later scan images. Inner hepatic vessels were compressed in 5 cases, vena cava and abdominal aortae were compressed in 3 cases. Pathological findings included fiber hyperplasia, hyaline degeneration, biliary duct hyperplasia, lobule-like array. CONCLUSION: MR imaging is a better way to differentiate and diagnose MHL. MHL may be recognized by its characteristic occurrence in infancy and MR imaging features. PMID- 16270390 TI - Microarray analysis of Escherichia coli O157:H7. AB - AIM: To establish the rapid, specific, and sensitive method for detecting O157:H7 with DNA microchips. METHODS: Specific oligonucleotide probes (26-28 nt) of bacterial antigenic and virulent genes of E. coli O157:H7 and other related pathogen genes were pre-synthesized and immobilized on a solid support to make microchips. The four genes encoding O157 somatic antigen (rfbE), H7 flagellar antigen (fliC) and toxins (SLT1, SLT2) were monitored by multiplex PCR with four pairs of specific primers. Fluorescence-Cy3 labeled samples for hybridization were generated by PCR with Cy3-labeled single prime. Hybridization was performed for 60 min at 45 degrees. Microchip images were taken using a confocal fluorescent scanner. RESULTS: Twelve different bacterial strains were detected with various combinations of four virulent genes. All the O157:H7 strains yielded positive results by multiplex PCR. The size of the PCR products generated with these primers varied from 210 to 678 bp. All the rfbE/fliC/SLT1/SLT2 probes specifically recognized Cy3-labeled fluorescent samples from O157:H7 strains, or strains containing O157 and H7 genes. No cross hybridization of O157:H7 fluorescent samples occurred in other probes. Non-O157:H7 pathogens failed to yield any signal under comparable conditions. If the Cy3-labeled fluorescent product of O157 single PCR was diluted 50-fold, no signal was found in agarose gel electrophoresis, but a positive signal was found in microarray hybridization. CONCLUSION: Microarray analysis of O157:H7 is a rapid, specific, and efficient method for identification and detection of bacterial pathogens. PMID- 16270391 TI - Enteroaggregative Escherichia coli isolated from Chinese diarrhea patients with high-pathogenicity island of Yersinia is involved in synthesis of siderophore yersiniabactin. AB - AIM: To investigate the distribution of 12 high-pathogenicity island (HPI) genes and the relation between HPI genes and expression of yersiniabactin (Ybt) in enteroaggregative E.coli (EAggEC) isolated from Chinese diarrhea patients. METHODS: The distribution of 12 HPI genes was investigated by PCR and DNA hybridization in two prototype strains of EAggEC, EAggEC 17-2, EAggEC O42, and 6 clinical EAggEC isolates from China. The production of siderophore Ybt in HPI positive strains was detected by reporter gene bioassay to determine the relation between HPI genes and expression of Ybt. Flow cytometry was used to detect fluorescent signal of the reporter strain that could designate production of Ybt. RESULTS: Seven strains were HPI-positive and one strain was HPI-negative. Six of seven HPI-positive strains were inserted into asnT-tRNA site. Moreover, seven EAggEC HPI-positive strains revealed enhanced fluorescence signal but the EAggEC HPI-negative strain did not. However, there was a difference in Ybt expression condition and level among these seven EAggEC HPI-positive strains. Although UFT073 strain, the prototype strain of uropathogenic E.coli (UPEC), carried the complete HPI core part, we did not detect the expression of Ybt in it. CONCLUSION: EAggEC HPI-positive strains can express the Ybt system, but the presence of HPI core part does not mean the functional expression of Ybt. PMID- 16270392 TI - Evaluation in vinyl chloride monomer-exposed workers and the relationship between liver lesions and gene polymorphisms of metabolic enzymes. AB - AIM: To analyze occupational health hazards exposure to doses lower than the Chinese occupational health standard in a selected VC polymerization plant in China, and also to elucidate the relationship between genetic polymorphisms and genetic susceptibility on liver lesions of workers exposed to vinyl chloride monomer(VCM). METHODS: In order to explore the mechanism of VCM-related health effects, we used a case-control design to investigate the association between the genetic polymorphisms of metabolic enzymes and liver lesions in workers occupationally exposed to VCM. Genotypes of CYP2E1, GSTT1, GSTM1, ALDH2 and ADH2 were identified using PCR and PCR-RFLP. RESULTS: Even when the concentration of VCM was lower than the current Chinese occupational health standard, neurasthenia, pharyngeal irritation, liver ultrasonography abnormalities and hemoglobin disorders were significantly higher in exposure subjects compared to non-exposure subjects, and the relative risks (RR and 95% CI) were 1.74 (1.06 2.85), 1.97 (1.56-2.48), 10.69 (4.38-26.12), and 2.07 (1.20-3.57). CYP2E1 c1c2/c2c2 genotype was significantly associated with liver damages (OR 3.29, 95% CI 1.51-7.20, P<0.01). CONCLUSION: The incidences of neurasthenia and liver ultrasonography abnormalities significantly increase when the cumulative exposure dose increases. The genotypes of metabolic enzymes (CYP2E1 c1c2/c2c2, null GSTT1 and ADH2 1-1) play important roles in VCM metabolism. Polymorphisms of CYP 2E1, GSTT1 and ADH2 may be a major reason of genetic susceptibility in VCM-induced hepatic damage. PMID- 16270393 TI - Strictures, diaphragms, erosions or ulcerations of ischemic type in the colon should always prompt consideration of nonsteroidal anti-inflammatory drug-induced lesions. AB - AIM: To investigate whether NSAIDs/ASA lesions in the colon can histologically be diagnosed on the basis of ischemic necrosis similar to biopsy-based diagnosis of NSAIDs/ASA-induced erosions and ulcers of the stomach. METHODS: In the period between 1997 and 2002, we investigated biopsy materials obtained from 611 patients (415 women, 196 men, average age 60.5 years) with endoscopic focal erosions, ulcerations, strictures or diaphragms in the colon. In the biopsies obtained from these lesions, we always established the suspected diagnosis of NSAID-induced lesions whenever necroses of the ischemic type were found. Together with the histological report, we enclosed a questionnaire to investigate the use of medication. The data provided by the questionnaire were then correlated with the endoscopic findings, the location, number and nature of the lesions, and the histological findings. RESULTS: At the time of their colonoscopy, 86.1% of the patients had indeed been taking NSAID/ASA medication for years (43.9%) or months (29.5%). The most common indication for the use of these drugs was pain (64.3%), and the most common indication for colonoscopy was bleeding (55.5%). Endoscopic inspection revealed multiple erosions and/or ulcers in 60.6%, strictures in 15.8%, and diaphragms in 3.0% of the patients. The lesions were located mainly in the right colon including the transverse colon (79.9%). A separate analysis of age and sex distribution, endoscopic and histological findings for NSAIDs alone, ASA alone, combined NSAID/ASA, and for patients denying the use of such drugs, revealed no significant differences among the groups. CONCLUSION: This uncontrolled retrospective study based on the histological finding of an ischemic necrosis shows that the histologically suspected diagnosis of NSAID-induced lesions in the colon is often correct. The true diagnostic validity of this finding and the differentiation from ischemic colitis should, however, be investigated in a prospective controlled study. PMID- 16270395 TI - Surgical risk for patients with Chagasic achalasia and its correlation with the degree of esophageal dilation. AB - AIM: To analyze the risk of cardiovascular complications in patients with indication for surgical treatment of Chagasic esophageal achalasia and to correlate the surgical risks with the degree of esophageal dilation, thereby proposing a risk scale index. METHODS: One hundred and twenty-four patients with Chagasic esophageal achalasia, who received surgical treatment at the Hospital das Clinicas of the Federal University of Goias, were included in this study. The patients were mostly related to the postoperative complications due to the cardiovascular system. All the patients were submitted to: (1) clinical history to define the cardiac functional class (New York Heart Association); (2) conventional 12-lead electrocardiogram at rest; and (3) contrast imaging of the esophagus to determine esophageal dilatation according to Rezende's classification of Chagasic megaesophagus. RESULTS: An assessment of the functional classification (FC) of heart failure during the preoperative period determined that 67 patients (54.03%) were assigned functional class I (FC I), 46 patients (37.09%) were assigned functional class II (FC II), and 11 patients (8.87%) were assigned functional class III (FC III). None of the patients were assigned to functional class IV (FC IV). There was a positive correlation between the functional class and the postoperative complications (FC IXFC II: P<0.001; FC IXFC III: P<0.001). The ECG was normal in 44 patients (35.48%) and presented abnormalities in 80 patients (64.52%). There was a significant statistical correlation between abnormal ECG (arrhythmias and primary change in ventricular repolarization) and postoperative complications (P<0.001). With regard to the classification of the Chagasic esophageal achalasia, the following distribution was observed: group II, 53 patients (42.74%); group III, 37 patients (29.83%); and group IV, 34 patients (27.41%). There was a positive correlation between the degree of esophageal dilation and the increase in postoperative complications (grade IIXgrade III achalasia: P<0.001; grade IIXgrade IV achalasia: P<0.001; and grade IIIXgrade IV achalasia: P = 0.017). Analyzing these results and using a multivariate regression analysis associated with the probability decision analysis, a risk scale was proposed as follows: up to 21 points (mild risk); from 22 to 34 points (moderate risk); and more than 34 points (high risk). The scale had 82.4% accuracy for mild risk patients and up to 94.6% for the high risk cases. CONCLUSION: The preoperative evaluation of the cardiovascular system, through a careful anamnesis, an ECG and contrast imaging of the esophagus, makes possible to estimate the surgical risks for Chagas' disease patients who have to undergo surgical treatment for esophageal achalasia. PMID- 16270394 TI - Association between thrombotic risk factors and extent of fibrosis in patients with non-alcoholic fatty liver diseases. AB - AIM: To evaluate the prevalence of genetic and acquired prothrombotic risk factors and their association with the extent of fibrosis and fatty infiltration in patients with non-alcoholic fatty liver disease (NAFLD). METHODS: Forty-four patients with chronic hepatitis (28 men and 16 women, with mean age of 45+/-11 and 49+/-12 years, respectively) constituted the patient population of this study. The groups were divided as follows: 15 patients with fatty liver (FL); 15 with non-alcoholic steatohepatitis (NASH); 14 with chronic viral hepatitis (CH) diagnosed by histology and liver technetium scan or ultrasound; and 10 healthy individuals. Thrombophilic, coagulation factors and genetic mutations were diagnosed by standard hemostatic and molecular coagulation assays. RESULTS: Activated protein C (APC) resistance and protein S were the most prevalent thrombotic risk factors (6% and 10% in NAFLD vs 21% and 14% in CH; P<0.01 and P<0.05, respectively). One thrombotic risk factor was identified in 41% of patients (23% mild fibrosis, 18% severe fibrosis) and two thrombotic risk factors in 6% of patients with NAFLD and severe fibrosis. While no differences in APC ratio, lupus anticoagulant, fibrinogen, factor V Leiden, prothrombin, and MTHFR mutation were found. Protein S levels were significantly lower in NASH patients than in patients with FL alone (92+/-19 vs 106+/-2, P<0.01). Protein C levels were markedly higher in patients with NAFLD and mild or severe fibrosis as compared to the patients with CH, respectively (128+/-40 vs 96+/-14, P<0.001 or 129+/-36 vs 88+/-13, P<0.01). CONCLUSION: Up to 46% of patients with NAFLD may have thrombotic risk factors, and the presence of thrombotic risk factors is correlated with the extent of hepatic fibrosis, suggesting a crucial role of the coagulation system in the pathogenesis of hepatic fibrosis. PMID- 16270396 TI - Focal adhesion kinase and Src phosphorylations in HGF-induced proliferation and invasion of human cholangiocarcinoma cell line, HuCCA-1. AB - AIM: To study the role of focal adhesion kinase (FAK) and its association with Src in hepatocyte growth factor (HGF)-induced cell signaling in cholangiocarcinoma progression. METHODS: Previously isolated HuCCA-1 cells were re-characterized by immunofluorescent staining and reverse transcriptase polymerase chain reaction assay for the expression of cytokeratin 19, HGF and c Met mRNA. Cultured HuCCA-1 cells were treated with HGF and determined for cell proliferation and invasion effects by MTT and invasion assays. Western blotting, immunoprecipitation, and co-immunoprecipitation were also performed to study the phosphorylation and interaction of FAK and Src. A novel Src inhibitor (AZM555130) was applied in cultures to investigate the effects on FAK phosphorylation inhibition and on cell proliferation and invasion. RESULTS: HGF enhanced HuCCA-1 cell proliferation and invasion by mediating FAK and Src phosphorylations. FAK Src interaction occurred in a time-dependent manner that Src was proved to be an upstream signaling molecule to FAK. The inhibitor to Src decreased FAK phosphorylation level in correlation with the reduction of cell proliferation and invasion. CONCLUSION: FAK plays a significant role in signaling pathway of HGF responsive cell line derived from cholangiocarcinoma. Autophosphorylated Src, induced by HGF, mediates Src kinase activation, which subsequently phosphorylates its substrate, FAK, and signals to cell proliferation and invasion. PMID- 16270397 TI - Effect of lysozyme chloride on betel quid chewing aggravated gastric oxidative stress and hemorrhagic ulcer in diabetic rats. AB - AIM: To evaluate the protective effect of lysozyme chloride on betel quid chewing (BQC) aggravated gastric oxidative stress and hemorrhagic ulcer in rats with diabetes mellitus (DM). METHODS: Male Wistar rats were challenged intravenously with streptozotocin (65 mg/kg) to induce DM. Rats were fed with regular pellet food or BQC-containing diets. After 90 d, rats were deprived of food for 24 h. Rat stomachs were irrigated for 3 h with normal saline or simulated gastric juice. Rats were killed and gastric specimens were harvested. RESULTS: An enhancement of various gastric ulcerogenic parameters, including acid back diffusion, mucosal lipid peroxide generation, as well as decreased glutathione levels and mucus content, were observed in DM rats. After feeding DM rats with BQC, an exacerbation of these ulcero-genic parameters was achieved. Gastric juice caused a further aggravation of these ulcerogenic parameters. Daily intragastric lysozyme chloride dose-dependently inhibited exacerbation of various ulcerogenic parameters in those BQC-fed DM rats. CONCLUSION: (1) Gastric juice could aggravate both DM and BQC-fed DM rat hemorrhagic ulcer; (2) BQC exacerbated gastric hemorrhagic ulcer in DM rats via enhancing oxidative stress and reducing defensive factors; (3) lysozyme chloride effectively protected BQC aggravated gastric damage in DM rats. PMID- 16270398 TI - Ulcerative colitis in a multiracial Asian country: racial differences and clinical presentation among Malaysian patients. AB - AIM: Inflammatory bowel disease appears to be uncommon among Asians. This study was conducted to determine the prevalence of ulcerative colitis (UC) in Malaysian patients and to establish the spectrum of the disease seen in Malaysian patients. Three major Asian races: Malay, Chinese, and Indian co-exist in Malaysia and we sought to determine if there were any racial differences in the prevalence and presentation of disease. Racial differences for several other gastrointestinal diseases have previously been observed and found to be extremely interesting. METHODS: Data were obtained retrospectively from a review of the medical records of in- and out-patients with a diagnosis of UC at the University Hospital, Kuala Lumpur between 1985 and 1998. RESULTS: There were 45 confirmed cases of UC of which 3 were foreigners, who were excluded from analysis. Thirty new cases of UC were diagnosed during the study period. Their mean age at presentation was 33.0+/ 10.0 years. The highest prevalence of UC was 17.9/100 000 hospital admissions in the Indians, followed by 11.2/100 000 hospital admissions in the Chinese. The lowest prevalence was 3.7/100 000 hospital admissions in the Malays. The prevalence of UC was significantly higher in the Indians and the Chinese when compared with the Malays with an OR of 4.89 (CI = 2.02-12.24; chi2 = 15.45, P<0.001) and 3.06 (CI = 1.24-7.78; chi2 = 6.30; P = 0.012) respectively. The extent of colonic disease was similar in the Malay and Indian patients. In contrast, distal or left-sided colitis predominated in the Chinese with an OR of 8.17 (95%CI = 1.31-64.87; chi2 = 5.53, P = 0.02). Extraintestinal manifestations were uncommon (11.9%). CONCLUSION: UC is an uncommon disease in Malaysia, but racial differences exist. The Indians had the highest prevalence of UC with the Chinese demonstrating the least extensive disease. PMID- 16270399 TI - Capsule endoscopy: improving transit time and image view. AB - AIM: To evaluate the effect of various methods of small bowel preparation on the transit time and the quality of visualization of the entire small bowel mucosa. METHODS: Ninety-five patients underwent capsule endoscopy (CE) by easily swallowing the capsule. They were divided into three study groups according to the preparation used: group A (n = 26) by polyethylene glycol (PEG) liter or with sodium phosphate (SP) 12 h prior to the CE study; group B (n = 29) by erythromycin 1 h prior to the CE study; and group C (n = 40) without any preparation. Visualization ranged from good to satisfactory to poor. RESULTS: The gastric emptying time in the group prepared with erythromycin was shorter but without statistical significance and the small bowel transit time was unaffected. In elderly subjects prepared by PEG or SP, the gastric emptying time was significantly longer (163.7 min, P = 0.05). The transit times of the three sub groups were not affected by gender or pathology. The grade of cleaning of the entire study group was 3.27+/-1.1. The erythromycin group presented significantly the worst quality of images (P = 0.05) compared to the other sub-groups. Age, gender, and pathology had no effect on the quality of the cleaning of the small bowel in the sub-groups. One (1.05%) case had no natural excretion. CONCLUSION: Erythromycin markedly reduces gastric emptying time, but has a negative effect on the quality of the image in the small bowel. The preparation of elderly subjects with PEG or SP has a negative effect on the small bowel transit time. PMID- 16270400 TI - Possible contribution of artificial neural networks and linear discriminant analysis in recognition of patients with suspected atrophic body gastritis. AB - AIM: To investigate whether ANNs and LDA could recognize patients with ABG in a database, containing only clinical and biochemical variables, of a pool of patients with and without ABG, by selecting the most predictive variables and by reducing input data to the minimum. METHODS: Data was collected from 350 consecutive outpatients (263 with ABG, 87 with non-atrophic gastritis and/or celiac disease [controls]). Structured questionnaires with 22 items (anagraphic, anamnestic, clinical, and biochemical data) were filled out for each patient. All patients underwent gastroscopy with biopsies. ANNs and LDA were applied to recognize patients with ABG. Experiment 1: random selection on 37 variables, experiment 2: optimization process on 30 variables, experiment 3: input data reduction on 8 variables, experiment 4: use of only clinical input data on 5 variables, and experiment 5: use of only serological variables. RESULTS: In experiment 1, overall accuracies of ANNs and LDA were 96.6% and 94.6%, respectively, for predicting patients with ABG. In experiment 2, ANNs and LDA reached an overall accuracy of 98.8% and 96.8%, respectively. In experiment 3, overall accuracy of ANNs was 98.4%. In experiment 4, overall accuracies of ANNs and LDA were, respectively, 91.3% and 88.6%. In experiment 5, overall accuracies of ANNs and LDA were, respectively, 97.7% and 94.5%. CONCLUSION: This preliminary study suggests that advanced statistical methods, not only ANNs, but also LDA, may contribute to better address bioptic sampling during gastroscopy in a subset of patients in whom ABG may be suspected on the basis of aspecific gastrointestinal symptoms or non-digestive disorders. PMID- 16270401 TI - Is adiponectin level a predictor of nonalcoholic fatty liver disease in nondiabetic male patients? AB - AIM: Adiponectin is a hepatic insulin sensitizer and also an inhibitor of tumor necrosis factor. We studied its levels in nondiabetic patients with nonalcoholic fatty liver disease (NAFLD) and compared with control group. METHODS: Thirty-five patients who had elevated serum aminotransferase levels with bright liver and 34 healthy volunteers without liver disease were evaluated. Age, gender and body mass index (BMI) were recorded. Fasting plasma glucose, insulin, adiponectin, proinsulin and lipid profile were measured. A standard oral glucose tolerance test (OGTT) with insulin response was performed and the index of insulin resistance was calculated according to the homeostasis model assessment (HOMA) method. RESULTS: According to the OGTT results, none of the participants had diabetes. Serum adiponectin levels were statistically significantly lower in patients with NAFLD than in control group (8.14+/-3.4 microg/mL vs 12.4+/-9.4 microg/mL, respectively, P<0.01). A statistically significant correlation was found between adiponectin and BMI (r : -0.33, P<0.01), HOMA (r : -0.26, P<0.05), proinsulin (r : -0.32, P<0.01), AST (r : -0.25, P<0.05), ALT (r : -0.26, P<0.05) or GGT (r : -0.22, P<0.05). In multiple regression analysis models, adiponectin levels were the only predictor of NAFLD in males, whereas in female group it was the BMI. CONCLUSION: Low adiponectin level might be a predictor of NAFLD especially in male nondiabetics. PMID- 16270402 TI - Comparison of a monoclonal antigen stool test (Hp StAR) with the 13C-urea breath test in monitoring Helicobacter pylori eradication therapy. AB - AIM: To evaluate the agreement between a mAb-based stool test (HP StAR) and the urea breath test (UBT) in monitoring (H pylori) infection after eradication therapy. METHODS: Patients with discordant results on UBT and Hp StAR underwent endoscopy with biopsies for rapid urease test, culture, and histology to confirm H pylori status. RESULTS: Among 250 patients (50+/-14 years), 240 (96.0%) had concordant UBT and Hp StAR tests with a significant correlation between DOB and A values (R = 0.87; P<0.0001). The remaining 10 (4.0%) patients had discordant tests (positive Hp StAR and negative UBT) with the Hp StAR inaccurate in five cases (false positive) and UBT inaccurate in the other five cases (false negative). The "maximal expected" sensitivity, specificity, +PV, -PV, +LR, and LR were 91%, 100%, 100%, 97.4%, infinity, and 8.2 respectively, for the UBT, and 100%, 97.4%, 91%, 100%, 38.8, and 0, respectively, for the Hp StAR. Overall accuracy for both tests was 98%. CONCLUSION: Both the UBT and the Hp StAR are equally accurate in monitoring H pylori infection. Nowadays, the choice of the "best" non-invasive H pylori test in the post-treatment setting should be done not only in terms of diagnostic accuracy but also in view of cost and local facilities. PMID- 16270403 TI - Interferon-alpha plus lamivudine vs lamivudine reduces breakthroughs, but does not affect sustained response in HBeAg negative chronic hepatitis B. AB - AIM: The HBeAg negative form of chronic hepatitis B (CHB) predominates in the Mediterranean area and has a rising frequency in Europe and North America. At present there are three approved therapies for patients with CHB: interferon alpha (IFN-alpha), lamivudine and adefovir dipivoxil. Unfortunately, none of these drugs are effective in achieving a sustained response in patients with HBeAg negative CHB. Therefore, new therapeutic approaches have been examined. Our aim was to investigate the efficacy of combination treatment of IFN-alpha and lamivudine compared to lamivudine monotherapy, after 24 mo of administration in HBeAg-negative hepatitis B patients. METHODS: Fifty consecutive patients were randomly assigned to receive IFN-alpha-2b (5 MU thrice per week, n = 24) plus lamivudine (100 mg daily) or lamivudine only (n = 26) for 24 mo. Patients were followed up for further 6 mo. The primary outcome was the proportion with sustained virological response (undetectable serum HBV DNA concentrations) and or sustained biochemical response (transaminase levels within normal range) at 30 mo (6 mo after the end of therapy). Secondary end-points were timed from initial virological (biochemical) response to VBR (BBR, respectively) and the emergence of YMDD mutants across the two arms. RESULTS: Five of twenty-four (21%) patients in the combination arm vs 3/26 (12%) in the lamivudine arm had sustained response (i.e., normal serum transaminase levels and undetectable HBV DNA by PCR assay) 6 mo after treatment discontinuation. A reduction in the emergence of YMDD mutants and in the development of virological breakthroughs was observed in patients receiving combination treatment (10% vs 46%, P = 0.01 and 14% vs 46%, P = 0.03, respectively). Time from initial virologic response to virologic breakthrough (VBR) was greater among initial responders receiving combination treatment compared to those receiving lamivudine (22.9 mo vs 15.9 mo, respectively; P = 0.005). CONCLUSION: Our results demonstrate that IFN-alpha plus lamivudine combination therapy did not increase the sustained response, compared to lamivudine. However, combination therapy reduces the likelihood of VBR due to YMDD mutants and prolongs the time period until the breakthrough development. PMID- 16270405 TI - Multidimensional assessment of neuro-psychiatric symptoms in patients with low grade hepatic encephalopathy: a clinical rating scale. AB - AIM: To evaluate the feasibility of a new clinical rating scale for a standardized assessment of cirrhosis-associated neuro-psychiatric symptoms. METHODS: Forty patients with liver cirrhosis (LC, with or without low-grade hepatic encephalopathy) were invest-igated using a clinical neuro-psychiatric rating scale based on a comprehensive list of neurological, psychomotor, cognitive, affective, behavioral symptoms, and symptoms of disturbed bioregulation. RESULTS: The analysis revealed that the majority of cirrhotic patients showed, besides characteristic neurological symptoms of hepatic encephalopathy, various psychomotor, affective and bioregulatory symptoms (disturbed sleep and sexual dysfunction). Patients were impaired in the following subscales: sleep and biorhythm disorder (75.0% of patients), Parkinsonoid symptoms (25.0%), affective symptoms (17.5%), and psychomotor retardation (12.5%). The increase of total neuro-psychiatric clinical score was significantly associated with the degree of hepatic enceph-alopathy. CONCLUSION: This study suggests that a substantial number of patients with LC and low-grade hepatic encephalopathy manifest various clinical neuro-psychiatric symptoms. The use of a rating scale, which explores clinical dimensions of hepatic encephalopathy, would improve the management of patients with LC. PMID- 16270404 TI - Low preoperative platelet counts predict a high mortality after partial hepatectomy in patients with hepatocellular carcinoma. AB - AIM: To assess the validity of our selection criteria for hepatectomy procedures based on indocyanine green disappearance rate (K(ICG)), and to unveil the factors affecting posthepatectomy mortality in patients with hepatocellular carcinoma (HCC). METHODS: A retrospective analysis of 198 consecutive patients with HCC who underwent partial hepatectomies in the past 14 years was conducted. The selection criteria for hepatectomy procedures during the study period were K(ICG)>or=0.12 for hemihepatectomy, K(ICG)>or=0.10 for bisegmentectomy, K(ICG)>or=0.08 for monosegmentectomy, and K(ICG)>or=0.06 for nonanatomic hepatectomy. The hepatectomies were categorized into three types: major hepatectomy (hemihepatectomy or a more extensive procedure), bisegmentectomy, and limited hepatectomy. Univariate (Fisher's exact test) and multivariate (the logistic regression model) analyses were used. RESULTS: Postoperative mortality was 5% after major hepatectomy, 3% after bisegmentectomy, and 3% after limited hepatectomy. The three percentages were comparable (P = 0.876). The platelet count of 7.3X10(4)/microL died of postoperative morbidity, whereas 25% (6/24 patients) of patients with a platelet count of cervix carcinoma HeLa (IC50 = 30.56 +/- 5.16 micromol), > melanoma Fem-x (IC50 = 32.17 +/- 5.75 micromol), > breast tumor MDA-MB-361 (IC50 = 35.04 +/- 2.95 micromol). In contrast, captopril, used in doses from 0.1-50 micromol, was ineffective (IC50 > 50 micromol) to the same cell lines. It is important to note that captopril in concentrations > 1 micromol led to a statistically significant increase in the percent of survived melanoma Fem-x cells (p < 0.05). Understanding the action of these established and clinically accepted agents could provide a basis for design of improved therapeutic regimens in the treatment of cancer diseases. PMID- 16270526 TI - High hRFI expression correlates with resistance to fluoropyrimidines in human colon cancer cell lines and in xenografts. AB - We previously reported that the over-expression of hRFI, a protein preferentially expressed in the digestive tract regions of several cancers, exhibited a tendency to inhibit TNF-alpha induced apoptosis. In this study, we sought to determine the potential effect of hRFI expression on the sensitivity to 5-fluorouracil (5-FU) and/or other fluoropyrimidines. For the whole lysates of 8 colon cancer cell lines, we performed Western blotting with anti-hRFI antibody and analyzed the correlations between the expression level of hRFI and the cell lines' sensitivity to 5-FU induced apoptosis. Furthermore, for a tissue microarray consisting of 32 xenograft derived human cancer cell lines, we examined the expression levels of hRFI and survivin by immunohistochemical staining, and analyzed the correlations between the expression of each protein and the sensitivity to several chemotherapeutic agents in the xenografts examined. Both in colon cancer cell lines and in xenografts, the expression level of hRFI was correlated with resistance to 5-FU and its derivatives. This evidence suggests that hRFI may be a marker predicting the response to fluorouracil derived chemotherapeutic agents and that the reduction of the expression level of hRFI might improve the outcome of chemotherapy. PMID- 16270527 TI - A study on the prognostic value of cyclins D1 and E expression levels in resectable gastric cancer and on some correlations between cyclins expression, histoclinical parameters and selected protein products of cell-cycle regulatory genes. AB - Cyclins condition the course of a cell cycle through the activation of appropriate serine-threonine kinases. Any variation in the cyclins' expression result in pathologies of the cell division, including neoplastic proliferation. Activity of the complexes of cyclins D1 and E with appropriate cyclin-dependent kinases may be inhibited by protein P21 (WAF1/CIP1) which functions as a cell growth cycle inhibitor. As yet, there have been rather few reports on the prognostic value of this cyclin expression assessment in gastric cancer, the kind of neoplasm still characterized by very poor prognosis. The study aimed at the assessment of expression levels of cyclins D1 and E in surgically removed gastric cancers, including the analysis of this prognostic value parameter, and attempted to determine some correlations between the expression of the examined cyclins and selected histoclinical and molecular parameters such as: patients' age and gender, histological type according to the Lauren classification, cancer stage (TNM), degree of histological malignancy (G) and level of expression of the cell cycle regulatory genes protein products--P53, P21, P27. Immunohistochemical analysis was performed on specimens obtained from radical stomach resections of 80 patients treated in the period 1992-1997 for gastric cancer stage I-IIIB (TNM UICC) at the Department of Surgical Oncology, Medical University of Lodz. For immunohistochemical examinations, the LSAB system was used, designed for assessment of antigen expression. In statistical analysis, Fisher's exact test was applied to evaluate correlations between the analyzed variables and Mantel Haenschel's test to evaluate their collinearity. For the evaluation of the effect of the analyzed variables on postoperation survival and recurrence-free survival the Cox regression model was used. When analyzing the prognostic value and survival period in association to the cyclins D1 and E expression levels, a statistically significant correlation was found only in relation to cyclin E expression: a survival period of minimum 5 year duration was significantly higher in the group displaying a negative, or only faintly positive, reaction to the presence of cyclin E, than in the group with a strongly positive response. Moreover, the analysis showed statistically significant non-linear dependence between the histological type of cancer in the Lauren classification as well as a degree of histological malignancy and the level of cyclin E expression, and a negative correlation between the level of cyclin E expression and the stage of cancer; In addition, a positive correlation between the level of P53 and cyclin E expression as well as statistically significant non-linear correlation between the level of cyclin E expression and the level of protein P21 expression was observed. However, no statistically significant correlations were found between the level of expression of the two cyclins and the level of protein P27 expression or between the levels of cyclin D1 and E expression in gastric cancer. Out of the two types of evaluated cyclins only cyclin E can be considered a significant regulatory factor and a useful prognostic parameter in gastric cancer. PMID- 16270528 TI - The gene expression level of transforming growth factor-beta (TGF-beta) as a biological prognostic marker of hepatocellular carcinoma. AB - The question that transforming growth factor-beta (TGF-beta) provides a tumor suppressive or a tumor promoting role is still unknown in hepatocellular carcinoma (HCC). In the present study, we quantitatively investigated the gene expression levels of TGF-beta in liver tissues from patients with HCC. We also evaluated the prognostic importance of TGF-beta gene in HCC patients. A total of 59 patients with primary HCC who underwent hepatectomy between 1993 and 2001 were enrolled. TGF-beta gene expression levels of tumors and of noncancerous livers were analyzed by real-time reverse transcriptase polymerase chain reaction (RT PCR). The percentage of apoptotic cells in tumor cells (apoptotic index: AI) was evaluated with immunohistochemistry. Also the expression of survivin protein (apoptosis inhibitor) in tumors was detected by immunohistochemistry. TGF-beta gene expression levels of tumors were compared with clinicopathological findings of patients. The relative expression level of TGF-beta mRNA of 59 tumor tissues did not differ from those of 8 normal liver tissues or 59 noncancerous liver tissues. The mean AI of 29 tumors with normal expression levels of TGF-beta gene (4%) was significantly higher than that of 30 tumors with low expression levels of TGF-beta gene (2.5%, p = 0.03). Thirteen out of 30 tumors (43%) with low expression level of TGF-P gene showed survivin positive, while only 4 out of 29 tumors (14%) with preserved expression of TGF-beta gene showed survivin positive. This difference was significant (p = 0.012). The overall 5-year survival rate of 29 patients with tumors with preserved TGF-beta gene prolonged to 72% compared with that of 30 patients who had tumors with suppressed TGF-beta gene (58%, p = 0.156). In HCC, TGF-beta gene may play a defensive role against tumor progression by regulating survivin protein expression and by controlling occurrence of spontaneous apoptosis in tumors. PMID- 16270529 TI - Clinical usefulness of CEA-mRNA determination in minor effusion. AB - Malignant pleural effusion of lung cancer is an important prognostic factor, even in minor effusions. Previous studies reported that cytological examination could not detect malignant cells in pleural dissemination cases. Therefore, we used real-time PCR as a more sensitive test to detect malignant cells. The subjects were selected from 132 primary lung cancer patients and 8 benign tumor patients as negative control. These subjects had no apparent pleural effusion or distant metastasis. All subjects were negative on cytological examination and without exfoliation evidence. The follow-up duration was 18.1 +/- 7.1 months (mean +/- SD). In the real-time PCR, the CEA-mRNA and GAPDH-mRNA parameters were measured simultaneously, and the CEA-mRNA ratio was obtained as normalized values of CEA mRNA divided by GAPDH-mRNA. The CEA-mRNA ratio in our study was correlated with lymph node metastasis (N-factor: p = 0.0948) and lymphatic invasion (Ly-factor: p = 0.0520). Using a proportional hazard model, with recurrence or death as terminal point, the CEA-mRNA ratio affected the recurrence risk by 1.920 (95% CI: 1.104-3.340) in Stage 1a. Using log rank testing, we found significant differences in the recurrence rate between the CEA-mRNA-positive and -negative cases (p = 0.0039) at cut-off point 0.1. PMID- 16270530 TI - Cell cycle proteins in laryngeal cancer: role in proliferation and prognosis. AB - A study of laryngeal carcinomas was performed in order to analyze (a) the expression of p53/p21, cyclin D1/cyclin E, p21/p27 (b) the relation of normal and abnormal protein expression, with the proliferation status, as determined by the expression of Ki67 and PCNA and (c) the correlation of our findings with prognosis. We performed a retrospective analysis of 57 cases of squamous cell carcinomas of the larynx. We applied monoclonal antibodies against p53, p21, p27, cyclin D1, cyclin E, Ki67 and PCNA, using streptavidin-biotin method. Analysis of the p53/p21 proteins, revealed abnormalities in 25/37 cases (67.57%), while 12/37 (32.43%) cases displayed normal phenotype (p53-/p21-). Analysis of cyclins revealed overexpression in 17/48 cases (35.42), while the majority 31/48(64.58%) displayed normal phenotype (cyclin D1-/cyclin E-). Concerning CDKIs expression, the majority 30/50(60%) presented high levels of both inhibitors (p21+/p27+). Cases with simultaneous overexpression of CDKIs demonstrated significantly higher levels of Ki67 protein (p = 0.05). Analysis of p53/p21, cyclin D/cyclin E, p21/p27 patterns showed no association between the presence of one or two alterations and prognosis. In conclusion, we demonstrated that p53 tumor suppressor pathway is frequently disrupted in laryngeal cancer. Furthermore, levels of CDKIs, although they act as cell cycle activity blockers, are not reliable markers for the estimation of laryngeal neoplastic cells growth fraction. PMID- 16270531 TI - The interplay between gastric cancer cell lines and PBMCs mediated by the CC chemokine RANTES plays an important role in tumor progression. AB - In the present study, we investigated the effect of the RANTES-mediated interaction between gastric carcinoma cell lines and peripheral blood mononuclear cells (PBMCs) in tumor progression. RANTES production in PBMCs stimulated by highly metastatic cancer cell line-conditioned supernatants was higher than in those stimulated by a less metastatic gastric cancer cell line-conditioned supernatant. RANTES receptors were expressed in PBMCs, but not in those cancer cell lines; therefore it was suggested that RANTES might affect PBMCs but not cancer cells. Matrix metalloproteinase (MMP)-9 expression in PBMCs was examined. Similar to RANTES production, MMP-9 expression in PBMCs stimulated by highly metastatic cell line-conditioned supernatants was higher than in that stimulated by a less metastatic cell line-conditioned supernatant. Invasion assays of gastric cancer cell lines were performed. Cancer cells cultured with PBMCs invaded into Matrigel more frequently than those without PBMCs. This invasive activity was highly inhibited by an anti-RANTES antibody. These results suggest that tumor cells can acquire the potential for invasion by cooperating with PBMCs and RANTES plays an important role in the interplay between tumor cells and PBMCs. It is thus thought that RANTES might be a candidate molecular target in the therapeutic strategy for gastric cancers. PMID- 16270532 TI - IFN-alpha enhances TNF-alpha-induced apoptosis through down-regulation of c-Myc protein expression in HL-60 cells. AB - We examined the effect of interferon-alpha (IFN-alpha) in combination with tumor necrosis factor-a (TNF-alpha) on growth, differentiation and apoptosis in HL-60 human myeloid leukemia cells. IFN-alpha inhibited cell growth and induced apoptosis, but not differentiation, in HL-60 cells. IFN-alpha enhanced TNF-alpha induced apoptosis. We also investigated the expression of c-Myc and Bcl-2 oncoproteins, which are implicated in the survival or death of a cell. The levels of c-Myc protein expression were not changed by IFN-alpha alone at 24hrs of treatment, but were down-regulated at 72 hrs, accompanied by the appearance of apoptotic cells. While, IFN-alpha did not affect the level of Bcl-2 protein expression during this cultivation time. Interestingly, a combination treatment of IFN-alpha with TNF-alpha showed a greater decrease of c-Myc expression than TNF-a alone at 24hrs. Whereas, IFN-alpha did not significantly modulate Bcl-2 expression levels which were down-regulated by TNF-alpha. Therefore, the enhancement of TNF-alpha-induced apoptosis by IFN-a might be closely associated with the greater down-regulation of c-Myc protein, rather than Bcl-2. In contrast, with rapid down-regulation of c-Myc expression caused by TNF-alpha, IFN alpha down-regulated c-Myc rather late (at 72 hrs), suggesting that both cytokines have a distinct pathway regulating c-Myc protein expression. However, the enhancement of apoptosis in the combination treatment would suggest the presence of a common signaling pathway for induction of apoptosis at down-stream of c-Myc. PMID- 16270533 TI - Level of 5-fluorodeoxyuridine 5'-monophosphate in cancerous tissue in patients with gastric cancer under preoperative administration of TS-1. A preliminary study. AB - Metabolizing enzymes such as thymidylate synthase (TS) and dihydropyrimidine dehydrogenase (DPD) have long been known to be useful for predicting response and outcome in patients receiving 5-fluorouracil (5-FU). However, few studies have examined the cancerous tissue levels of 5-fluorodeoxyuridine 5'-monophosphate (FdUMP), a metabolite of 5-FU that has an important role in inhibiting DNA synthesis. In this study, for the first time to our knowledge, we measured concentrations of FdUMP in tumor specimens and surrounding non-cancerous tissue obtained at operation in 10 patients with gastric cancer who received TS-1 before surgery (80 mg/m2, 3 days). The FdUMP level in the cancerous tissue was significantly higher than that in the non-cancerous tissue (153.0 +/- 85.7 pmol/g tissue vs. 53.0 +/- 47.0 pmol/g tissue)(p = 0.0046). Furthermore, the TS level in tumor was significantly higher than that in non-cancerous tissue (6.362 +/- 5.106 pmol/g tissue vs. 2.092 +/- 2.050 pmol/g tissue) (p = 0.0310). The mean ratios of TS-bound FdUMP to TS and FdUMP concentrations in the cancerous tissues were 45.9% and 2.00%, respectively. Our results demonstrate that in cancerous tissue, TS-1 may produce high FdUMP concentration and suppress about half FdUMP concentration by forming ternary complexes. PMID- 16270534 TI - Effect of sodium butyrate on pro-matrix metalloproteinase-9 and -2 differential secretion in pediatric tumors and cell lines. AB - Matrix metalloproteinases (MMPs) are enzymes responsible for extracellular matrix degradation and contribute to local and distant cell invasion during cancer progression or metastasis. The effects of chromatin structure on gene expression and the use of histone deacetylase inhibitors such as sodium butyrate (NaBu) may directly influence pro-MMPs secretion. In the present study, we evaluated the effect of NaBu on pro-MMP-9 and pro-MMP-2 secretion in human Jurkat and HT1080 cells, and in 36 pediatric solid tumors. Cell lines and samples were exposed to 8 mM of NaBu and proteinase activity was evaluated in the supernatant by gelatin zymograms. Our results showed, for Jurkat cells treated with NaBu, increases of 2 fold and 1.5-fold in pro-MMP-9 and pro-MMP-2 secretion, respectively. A 50% decrease in pro-MMP-9 secretion due to NaBu was observed in HT1080 cells. NaBu induced a 0.62 reduction in levels of pro-MMP-9 secretion in untreated tumors. For cell lines and some NaBu-treated tumors we found histone H4 hyperacetylation. We conclude that pro-MMPs gene expression and their secretion can be epigenetically mis-regulated in tumoral processes. PMID- 16270535 TI - Experimental study on pathogenesis and histomorphology of early carcinoma of the extrahepatic bile duct in the Syrian hamster. AB - To elucidate the pathogenesis of carcinomas in the extrahepatic bile duct, we investigated the histomorphological characteristics of adenomas and early carcinomas induced in the extrahepatic bile duct of hamsters. Syrian hamsters underwent a cholecystoduodenostomy along with a dissection of the common duct, while also being administered N-nitrosobis(2-oxopropyl)amine (BOP). The tumors that arose from the extrahepatic bile duct included 10 adenomas and 55 early carcinomas in 56 of the 156 hamsters sacrificed. All the adenomas were found to be polypoid in shape. The early carcinomas, which were restricted within the mucosal layer of the bile duct, showed the following three different growth patterns: (1) protruding type in 41 (75%), consisting of 27 polypoid and 14 papillary tumors; (2) superficial spreading type in 9 (16%); and (3) periductal glandular type in 5 (9%). There were no depressed tumors observed. Carcinomas existing either alone or associated with adenomas were evident in 12 (22%) tumors, and 11 of these were polypoid. Atypical papillary hyperplasia within the tumor mass was noted in 22 early carcinomas (40%) and was particularly prominent in papillary type tumors. These results support the concept of an adenoma carcinoma sequence in the majority of polypoid tumors of the extrahepatic bile duct. Atypical papillary hyperplasia might also be premalignant, and these precursor lesions should reflect the growth patterns of tumors, at least in the early stage of tumorigenesis. PMID- 16270536 TI - Cerebellar atrophy and monoclonal gammopathy of undetermined significance: a possible correlation? AB - Central Nervous System involvement in Monoclonal gammopathies of undetermined significance has seldomly been reported and in all the cases a demyelinating disease was found. We report the case of a young man who had been suffering for five years of progressive cerebellar syndrome. MRI showed marked cerebellar vermis atrophy. An IgG lambda monoclonal gammopathy was revealed in the serum. Cerebrospinal fluid examination showed oligoclonal bands and elevated Link-Index. Serologic research for HBV, HCV, HIV, Lues, Rubella, Measles was negative, as also genetic analysis for SCA1, SCA2, SCA3, SCA7 and Friederich's ataxia. Nerve conduction studies were normal. Plasmatic vit.E was low, but treatment with high doses of tocopherol was ineffective. i.v. immunoglobulins and steroids obtained only transient clinical benefits. In conclusion, we hypothesize a pathogenetic role of the IgG in this cerebellar atrophy. PMID- 16270537 TI - Concomitant leiomyoma and leiomyosarcoma of the oesophagus. AB - Leiomyosarcoma of the oesophagus is a malignant tumor that originates from smooth muscle cells. The filiation between oesophageal leiomyoma and leiomyosarcoma is controversial, with few cases reported in literature. The authors describe un uncommon situation with the simultaneous presence of a leiomyoma and a leiomyosarcoma of the oesophagus in a 75 year-old man, which have been successfully treated with surgical resection. It could probably be a case of a malignant transformation of a previous pre-existing multiple non-confluent leiomyoma. PMID- 16270539 TI - Cutaneous lymphoadenoma: a rare clinicopathological entity. AB - Cutaneous Lymphadenoma (Benign Lymphoepithelial tumour of the skin) is a rare tumour, with distinctive clinical and histological features. To date, very few cases of this entity have been reported. We present a case of cutaneous lymphoadenoma in a 52-year-old man and a short review of the literature, summarizing the principal clinical and morphological characteristics of this rare tumour. PMID- 16270538 TI - Choroid plexus carcinoma in the adult: is there a role for chemotherapy? AB - Choroid plexus carcinoma is a rare primary brain neoplasm arising from epithelial differentiated tissue, originating from the choroids plexus of the ventricles and, in 90% of the cases, in the lateral and fourth ventricles. This neoplasm is seen mainly in children and reported infrequently in adults. The treatment of choroid plexus carcinoma is based on scarce evidence in literature. We report a rare case of an adult woman affected by a choroid plexus tumour and a discussion on the therapeutic management of this uncommon adult malignancy. PMID- 16270540 TI - Laser diagnostics for gas dynamics: introduction to the feature issue. PMID- 16270541 TI - Time-resolved CO2 thermometry for pressures as great as 5 MPa by use of pure rotational coherent anti-Stokes Raman scattering. AB - Pure rotational coherent anti-Stokes Raman scattering measurements of pure CO2 have been performed in a temperature range from 300 to 773 K and for pressure from 0.1 to 5 MPa for the purpose of time-resolved CO2 thermometry. Particular emphasis was put on the comparison of several line-width approximations to model the experimental spectra. Generally good agreement of the temperature mean values with the thermocouple reference has been found for all models over almost the whole pressure and temperature range investigated. The standard deviations, which increased with temperature, were comparable with or better than the results gained for single-shot measurements of pure N2 or O2-N2 mixtures. Yet for high particle densities close to the critical point of CO2 the limitation of the models became obvious, owing to the strongly increased influence of motional narrowing effects. The characteristics of these effects have been demonstrated by measurements even closer to the critical conditions. PMID- 16270543 TI - Simple dense-pattern optical multipass cells. AB - Multiple-pass optical cells with dense spot patterns are useful for many applications, especially when the cell volume must be minimized relative to the optical path length. Present methods to achieve these dense patterns require expensive, highly precise astigmatic mirrors and complex alignment procedures. This work describes a new, simpler, and less demanding mirror system, comprising either a pair of cylindrical mirrors or one cylindrical and one spherical mirror. PMID- 16270544 TI - Use of Rayleigh imaging and ray tracing to correct for beam-steering effects in turbulent flames. AB - Laser Rayleigh imaging has been applied in a number of flow and flame studies to measure concentration or temperature distributions. Rayleigh cross sections are dependent on the index of refraction of the scattering medium. The same index of refraction changes that provide contrast in Rayleigh images can also deflect the illuminating laser sheet. By applying a ray-tracing algorithm to the detected image, it is possible to correct for some of these beam-steering effects and thereby improve the accuracy of the measured field. Additionally, the quantification of the degree of beam steering through the flow provides information on the degradation of spatial resolution in the measurement. Application of the technique in a well-studied laboratory flame is presented, along with analysis of the effects of image noise and spatial resolution on the effectiveness of the algorithm. PMID- 16270542 TI - Soot particle disintegration and detection by two-laser excimer laser fragmentation fluorescence spectroscopy. AB - A two-laser technique is used to study laser-particle interactions and the disintegration of soot by high-power UV light. Two separate 20 ns laser pulses irradiate combustion-generated soot nanoparticles with 193 nm photons. The first laser pulse, from 0 to 14.7 J/cm2, photofragments the soot particles and electronically excites the liberated carbon atoms. The second laser pulse, held constant at 13 J/cm2, irradiates the remaining particle fragments and other products of the first laser pulse. The atomic carbon fluorescence at 248 nm produced by the first laser pulse increases linearly with laser fluence from 1 to 6 J/cm2. At higher fluences the signal from atomic carbon saturates. The carbon fluorescence from the second laser pulse decreases as the fluence from the first laser increases, suggesting that the particles fully disintegrate at high laser fluences. We use an energy balance parameter, called the photon/atom ratio, to aid in understanding laser-particle interactions. These results help define the regimes where photofragmentation fluorescence methods quantitatively measure total soot concentrations. PMID- 16270545 TI - Phase-locked two-line OH planar laser-induced fluorescence thermometry in a pulsating gas turbine model combustor at atmospheric pressure. AB - Two-line OH planar laser-induced fluorescence (PLIF) thermometry was applied to a swirling CH4/air flame in a gas turbine (GT) model combustor at atmospheric pressure, which exhibited self-excited combustion instability. The potential and limitations of the method are discussed with respect to applications in GT-like flames. A major drawback of using OH as a temperature indicator is that no temperature information can be obtained from regions where OH radicals are missing or present in insufficient concentration. The resulting bias in the average temperature is addressed and quantified for one operating condition by a comparison with results from laser Raman measurements applied in the same flame. Care was taken to minimize saturation effects by decreasing the spectral laser power density to a minimum while keeping an acceptable spatial resolution and signal-to-noise ratio. In order to correct for the influence of laser light attenuation, absorption measurements were performed on a single-shot basis and a correction procedure was applied. The accuracy was determined to 4%-7% depending on the location within the flame and on the temperature level. A GT model combustor with an optical combustion chamber is described, and phase-locked 2D temperature distributions from a pulsating flame are presented. The temperature variations during an oscillation cycle are specified, and the general flame behavior is described. Our main goals are the evaluation of the OH PLIF thermometry and the characterization of a pulsating GT-like flame. PMID- 16270546 TI - Toward in-cylinder absorption tomography in a production engine. AB - Design requirements for an 8000 frame/s dual-wavelength ratiometric chemical species tomography system, intended for hydrocarbon vapor imaging in one cylinder of a standard automobile engine, are examined. The design process is guided by spectroscopic measurements on iso-octane and by comprehensive results from laboratory phantoms and research engines, including results on temporal resolution performance. Novel image reconstruction techniques, necessary for this application, are presented. Recent progress toward implementation, including details of the optical access arrangement employed and signal-to-noise issues, is described. We present first cross-cylinder IR absorption measurements from a reduced channel-count (nontomographic) system and discuss the prospects for imaging. PMID- 16270547 TI - Detection of high-temperature water vapor at 940 nm with vertical-cavity surface emitting lasers. AB - A vertical-cavity surface-emitting laser was used to study the absorption of water vapor in the 940 nm region. Measurements for several absorption lines within the 2 v1 + V3 vibrational band were performed. Line strengths at room temperature and in a heated absorption cell over the temperature range of 420-970 K were obtained. The line strength values were in good agreement with simulations based on the values of the HITRAN 2004 database. The measurements also showed that water vapor transitions near 940 nm are suitable for sensitive temperature determination. PMID- 16270548 TI - Temperature measurement using ultraviolet laser absorption of carbon dioxide behind shock waves. AB - A diagnostic for microsecond time-resolved temperature measurements behind shock waves, using ultraviolet laser absorption of vibrationally hot carbon dioxide, is demonstrated. Continuous-wave laser radiation at 244 and 266 nm was employed to probe the spectrally smooth CO2 ultraviolet absorption, and an absorbance ratio technique was used to determine temperature. Measurements behind shock waves in both nonreacting and reacting (ignition) systems were made, and comparisons with isentropic and constant-volume calculations are reported. PMID- 16270550 TI - Hydroxyl tagging velocimetry method optimization: signal intensity and spectroscopy. AB - The previously demonstrated nonintrusive time-of-flight molecular velocity tagging method, hydroxyl tagging velocimetry (HTV), has shown the capability of operating both at room temperature and in flames. Well-characterized jets of either air (nonreacting cases) or hydrogen-air diffusion flames (reacting cases) are employed. A 7 x 7 OH line grid is generated first through the single-photon photodissociation of H2O by a approximately 193 nm pulsed narrowband ArF excimer laser and is subsequently revealed by a read laser sheet through fluorescence caused by A2sigma+(v' = 3) <-- X2pi(i)(v'' = 0), A2sigma+(v' = 1) <-- X2pi(i)(v'' = 0), or A2sigma+(v' = 0) < or = X2pi(i)(v'' = 0) pumping at approximately 248, approximately 282, or approximately 308 nm, respectively. A detailed discussion of the spectroscopy and relative signal intensity of these various read techniques is presented, and the implications for optimal HTV performance are discussed. PMID- 16270549 TI - Quantitative analysis of the near-wall mixture formation process in a passenger car direct-injection diesel engine by using linear raman spectroscopy. AB - Optimum fuel preparation and mixture formation are core issues in the development of modern direct-injection (DI) Diesel engines, as these are crucial for defining the border conditions for the subsequent combustion and pollutant formation process. The local fuel/air ratio can be seen as one of the key parameters for this optimization process, as it allows the characterization and comparison of the mixture formation quality. For what is the first time to the best of our knowledge, linear Raman spectroscopy is used to detect the fuel/air ratio and its change along a line of a few millimeters directly and nonintrusively inside the combustion bowl of a DI Diesel engine. By a careful optimization of the measurement setup, the weak Raman signals could be separated successfully from disturbing interferences. A simultaneous measurement of the densities of air and fuel was possible along a line of about 10 mm length, allowing a time- and space resolved measurement of the local fuel/air ratio. This could be performed in a nonreacting atmosphere as well as during fired operating conditions. The positioning of the measurement volume next to the interaction point of one of the spray jets with the wall of the combustion bowl allowed a near-wall analysis of the mixture formation process for a six-hole nozzle under varying injection and engine conditions. The results clearly show the influence of the nozzle geometry and preinjection on the mixing process. In contrast, modulation of the intake air temperature merely led to minor changes of the fuel concentration in the measurement volume. PMID- 16270551 TI - Ballistic imaging of the liquid core for a steady jet in crossflow. AB - A time-gated ballistic imaging instrument is used to obtain high-spatial resolution, single-shot images of the liquid core in a water spray issuing into a gaseous crossflow. We describe further development of the diagnostic technique to improve spatial resolution and present images and statistics for various jets under crossflow experimental conditions (different Weber numbers). Series of these images reveal a near-nozzle flow field undergoing breakup and subsequent droplet formation by stripping. One can also detect signatures of spatially periodic behavior in the liquid core and formation of small voids during breakup. PMID- 16270552 TI - Quantum cascade laser sensor for SO2 and SO3 for application to combustor exhaust streams. AB - We have demonstrated a high-sensitivity, room-temperature quantum-cascade (QC) laser sensor for detection of SO2 and SO3 under conditions relevant to aircraft test combustor exhaust. Two QC lasers probe infrared absorption features at 7.50 and 7.16 microm for SO2 and SO3, respectively, with a common dual-beam detection system. We inferred a noise-equivalent absorbance of approximately 1 x 10(-4) Hz( 1/2). We have demonstrated detection limits for both SO2 and SO3 of 1-2 ppmv m/Hz(1/2) (where ppmv is parts in 10(6) by volume) for 300 torr, elevated temperature, and path lengths near 1 m. This level of sensitivity permits measurement of < 1 ppmv of SO2 and SO3 at these conditions with modest signal averaging. PMID- 16270553 TI - Two-photon laser-induced fluorescence of atomic hydrogen in a diamond-depositing dc arcjet. AB - Atomic hydrogen in the plume of a dc-arcjet plasma is monitored by use of two photon excited laser-induced fluorescence (LIF) during the deposition of diamond film. The effluent of a dc-arc discharge in hydrogen and argon forms a luminous plume as it flows through a converging-diverging nozzle into a reactor. When a trace of methane (< 2%) is added to the flow in the diverging part of the nozzle, diamond thin film grows on a water-cooled molybdenum substrate from the reactive mixture. LIF of atomic hydrogen in the arcjet plume is excited to the 3S and 3D levels with two photons near 205 nm, and the subsequent fluorescence is observed at Balmer-alpha near 656 nm. Spatially resolved LIF measurements of atomic hydrogen are made as a function of the ratio of hydrogen to argon feedstock gas, methane addition, and reactor pressure. At lower reactor pressures, time-resolved LIF measurements are used to verify our collisional quenching correction algorithm. The quenching rate coefficients for collisions with the major species in the arcjet (Ar, H, and H2) do not change with gas temperature variations in the plume (T < 2300 K). Corrections of the LIF intensity measurements for the spatial variation of collisional quenching are important to determine relative distributions of the atomic hydrogen concentration. The relative atomic hydrogen concentrations measured here are calibrated with an earlier calorimetric determination of the feedstock hydrogen dissociation to provide quantitative hydrogen-atom concentration distributions. PMID- 16270554 TI - Diode laser absorption spectroscopy of water vapor in a scramjet combustor. AB - A sensor based on tunable diode laser absorption spectroscopy was constructed for time-resolved temperature and water vapor concentration measurements in a scramjet combustor. The sensor probed two absorption lines near 1390 nm with two time-multiplexed lasers used to measure temperature and water vapor concentration at up to 20 kHz. A demonstration experiment was performed in the supersonic, expanding exhaust region of the combustor, showing the measurement to be repeatable, able to resolve temporal trends during tunnel operation, and sensitive to changes in combustor operating conditions. PMID- 16270555 TI - Quantitative determination of combustion intermediates with cavity ring-down spectroscopy: systematic study in propene flames near the soot-formation limit. AB - Cavity ring-down spectroscopy (CRDS) was applied in several fuel-rich, one dimensional, premixed C3H6/O2/Ar flames at 50 mbars (37.5 torr) to measure absolute OH, HCO, and 1CH2 concentration as well as temperature as a function of stoichiometry. Although these flames near the sooting limit present a complex chemical environment, significant spectral interferences were found to be absent. Specific aspects of the CRDS technique for measurement of temperature and radical concentration profiles are discussed; and the results are analyzed in comparison with flame model simulations. PMID- 16270556 TI - Soot formation and oxidation in oscillating methane-air diffusion flames at elevated pressure. AB - Comparisons with respect to the sooting tendency are made between stationary diffusion flames and diffusion flames with pulsations induced by oscillating fuel flow. Time-resolved measurements of the soot particle properties in the flames are obtained by combining Rayleigh-scattering, laser-induced incandescence, and extinction measurements into the RAYLIX method. Furthermore, flame luminosity at 590 nm and OH*-chemoluminescence signals at 310 nm are monitored to obtain data regarding the flame structure. Mean soot volume fractions of oscillating flames are significantly different from those of stationary flames with the same mean fuel flow rate; oscillations of the total amount of soot are phase shifted and asymmetric compared with fuel flow oscillations. PMID- 16270557 TI - High-speed fuel tracer fluorescence and OH radical chemiluminescence imaging in a spark-ignition direct-injection engine. AB - An innovative technique has been demonstrated to achieve crank-angle-resolved planar laser-induced fluorescence (PLIF) of fuel followed by OH* chemiluminescence imaging in a firing direct-injected spark-ignition engine. This study used two standard KrF excimer lasers to excite toluene for tracking fuel distribution. The intensified camera system was operated at single crank-angle resolution at 2000 revolutions per minute (RPM) for 500 consecutive cycles. Through this work, it has been demonstrated that toluene and OH* can be imaged through the same optical setup while similar signal levels are obtained from both species, even at these high rates. The technique is useful for studying correlations between fuel distribution and subsequent ignition and flame propagation without the limitations of phase-averaging imaging approaches. This technique is illustrated for the effect of exhaust gas recirculation on combustion and will be useful for studies of misfire causes. Finally, a few general observations are presented as to the effect of preignition fuel distribution on subsequent combustion. PMID- 16270558 TI - Hydroxyl tagging velocimetry in a supersonic flow over a cavity. AB - Hydroxyl tagging velocimetry (HTV) measurements of velocity were made in a Mach 2 (M 2) flow with a wall cavity. In the HTV method, ArF excimer laser (193 nm) beams pass through a humid gas and dissociate H2O into H + OH to form a tagging grid of OH molecules. In this study, a 7 x 7 grid of hydroxyl (OH) molecules is tracked by planar laser-induced fluorescence. The grid motion over a fixed time delay yields about 50 velocity vectors of the two-dimensional flow in the plane of the laser sheets. Velocity precision is limited by the error in finding the crossing location of the OH lines written by the excimer tag laser. With a signal to-noise ratio of about 10 for the OH lines, the determination of the crossing location is expected to be accurate within +/- 0.1 pixels. Velocity precision within the freestream, where the turbulence is low, is consistent with this error. Instantaneous, single-shot measurements of two-dimensional flow patterns were made in the nonreacting M 2 flow with a wall cavity under low- and high pressure conditions. The single-shot profiles were analyzed to yield mean and rms velocity profiles in the M 2 nonreacting flow. PMID- 16270559 TI - Near-infrared diode laser absorption diagnostic for temperature and water vapor in a scramjet combustor. AB - Tunable diode laser absorption measurements of gas temperature and water concentration were made at the exit of a model scramjet combustor fueled on JP-7. Multiplexed, fiber-coupled, near-infrared distributed feedback lasers were used to probe three water vapor absorption features in the 1.34-1.47 microm spectral region (2v1 and vl + v3 overtone bands). Ratio thermometry was performed using direct-absorption wavelength scans of isolated features at a 4-kHz repetition rate, as well as 2f wavelength modulation scans at a 2-kHz scan rate. Large signal-to-noise ratios demonstrate the ability of the optimally engineered optical hardware to reject beam steering and vibration noise. Successful measurements were made at full combustion conditions for a variety of fuel/air equivalence ratios and at eight vertical positions in the duct to investigate spatial uniformity. The use of three water vapor absorption features allowed for preliminary estimates of temperature distributions along the line of sight. The improved signal quality afforded by 2f measurements, in the case of weak absorption, demonstrates the utility of a scanned wavelength modulation strategy in such situations. PMID- 16270560 TI - Cavity-enhanced quantum-cascade laser-based instrument for carbon monoxide measurements. AB - An autonomous instrument based on off-axis integrated cavity output spectroscopy has been developed and successfully deployed for measurements of carbon monoxide in the troposphere and tropopause onboard a NASA DC-8 aircraft. The instrument (Carbon Monoxide Gas Analyzer) consists of a measurement cell comprised of two high-reflectivity mirrors, a continuous-wave quantum-cascade laser, gas sampling system, control and data-acquisition electronics, and data-analysis software. CO measurements were determined from high-resolution CO absorption line shapes obtained by tuning the laser wavelength over the R(7) transition of the fundamental vibration band near 2172.8 cm(-1). The instrument reports CO mixing ratio (mole fraction) at a 1-Hz rate based on measured absorption, gas temperature, and pressure using Beer's Law. During several flights in May-June 2004 and January 2005 that reached altitudes of 41,000 ft (12.5 km), the instrument recorded CO values with a precision of 0.2 ppbv (1-s averaging time) and an accuracy limited by the reference CO gas cylinder (uncertainty < 1.0%). Despite moderate turbulence and measurements of particulate-laden airflows, the instrument operated consistently and did not require any maintenance, mirror cleaning, or optical realignment during the flights. PMID- 16270561 TI - Quantitative temperature measurements in high-pressure flames with multiline NO LIF thermometry. AB - An accurate temperature measurement technique for steady, high-pressure flames is investigated using excitation wavelength-scanned laser-induced fluorescence (LIF) within the nitric oxide (NO) A-X(0, 0) band, and demonstration experiments are performed in premixed methane/air flames at pressures between 1 and 60 bars with a fuel/air ratio of 0.9. Excitation spectra are simulated with a computational spectral simulation program (LIFSim) and fit to the experimental data to extract gas temperature. The LIF scan range was chosen to provide sensitivity over a wide temperature range and to minimize LIF interference from oxygen. The fitting method is robust against elastic scattering and broadband LIF interference from other species, and yields absolute, calibration-free temperature measurements. Because of loss of structure in the excitation spectra at high pressures, background signal intensities were determined using a NO addition method that simultaneously yields nascent NO concentrations in the postflame gases. In addition, fluorescence emission spectra were also analyzed to quantify the contribution of background signal and to investigate interference in the detection band-width. The NO-LIF temperatures are in good agreement with intrusive single-color pyrometry. The proposed thermometry method could provide a useful tool for studing high-pressure flame chemistry as well as provide a standard to evaluate and validate fast-imaging thermometry techniques for practical diagnostics of high-pressure combustion systems. PMID- 16270562 TI - Measurements of OH mole fraction and temperature up to 20 kHz by using a diode laser-based UV absorption sensor. AB - Diode-laser-based sum-frequency generation of ultraviolet (UV) radiation at 313.5 nm was utilized for high-speed absorption measurements of OH mole fraction and temperature at rates up to 20 kHz. Sensor performance was characterized over a wide range of operating conditions in a 25.4 mm path-length, steady, C2H4-air diffusion flame through comparisons with coherent anti-Stokes Raman spectroscopy (CARS), planar laser-induced fluorescence (PLIF), and a two-dimensional numerical simulation with detailed chemical kinetics. Experimental uncertainties of 5% and 11% were achieved for measured temperatures and OH mole fractions, respectively, with standard deviations of < 3% at 20 kHz and an OH detection limit of < 1 part per million in a 1 m path length. After validation in a steady flame, high-speed diode-laser-based measurements of OH mole fraction and temperature were demonstrated for the first time in the unsteady exhaust of a liquid-fueled, swirl stabilized combustor. Typical agreement of approximately 5% was achieved with CARS temperature measurements at various fuel/air ratios, and sensor precision was sufficient to capture oscillations of temperature and OH mole fraction for potential use with multiparameter control strategies in combustors of practical interest. PMID- 16270563 TI - Two-point, high-repetition-rate Rayleigh thermometry in flames: techniques to correct for apparent dissipation induced by noise. AB - High-repetition-rate, two-point Rayleigh thermometry is used to measure the thermal dissipation in turbulent nonpremixed jet flames. Scalar-dissipation measurements are very important in turbulent combustion but are often strongly influenced by noise effects. Dissipation is proportional to the squared gradient of the scalar, and noise produces an "apparent dissipation" that can dominate the measured dissipation, particularly at high resolution. Two independent techniques are presented that enable correction for the apparent thermal dissipation, provided that the smallest spatial scales are resolved. A model for shot-noise limited data is developed that predicts the magnitude of the apparent dissipation at any measurement location and gives the minimum value of the apparent dissipation for measurements that are not shot-noise limited. These techniques are applied to the Rayleigh thermometry data, and they are shown to be largely self-consistent and consistent with theoretical expectations. The apparent dissipation is significantly larger than the true dissipation, demonstrating the importance of data correction in this noise-limited, fully spatially resolved regime. PMID- 16270564 TI - Continuous-wave cavity ringdown absorption spectroscopy with a swept-frequency laser: rapid spectral sensing of gas-phase molecules. AB - A cavity ringdown spectrometer, based on a continuous-wave swept-frequency laser, enables efficient, rapid recording of wide-ranging absorption spectra as characteristic spectral signatures of airborne molecules. The rapidly swept laser frequency resonates with the longitudinal modes of the ringdown cavity, effectively sampling the absorption spectrum of an intracavity gas at intervals defined by the cavity's free spectral range and generating a full absorption spectrum within a single rapid sweep of the widely tunable laser frequency. We report a new analog detection scheme that registers a single data point for each buildup and ringdown decay event without logging details of the full signal waveform; this minimizes demand on digitizer speed and memory depth, reducing the time scale of data processing. This results in a compact, robust, easy-to-use instrument that offers fresh prospects for spectroscopic sensing of trace species in the atmosphere. PMID- 16270565 TI - Optical design in beam steering environments with emphasis on laser transmission measurements. AB - Optical sensors applied to practical devices often encounter beam steering: the wander and/or diffusion of laser light. Here we provide a framework for minimizing the sensitivity of transmission-based sensors to beam steering without quantitative prediction of the severity of the beam-steering field. Typical goals are increased transmission and/or minimized fluctuations in transmission; such features can improve optical sensor performance (e.g., improved signal-to-noise ratio, response time, or spectral resolution). In our framework, we introduce a parameter for characterizing beam-steering severity. We then compare two approaches for absorption spectroscopy and show that the preferred approach depends on the total spectral range monitored, the spectral resolution desired, and the severity of the beam steering. PMID- 16270566 TI - A calibration-independent laser-induced incandescence technique for soot measurement by detecting absolute light intensity. AB - Laser-induced incandescence (LII) has proved to be a useful diagnostic tool for spatially and temporally resolved measurement of particulate (soot) volume fraction and primary particle size in a wide range of applications, such as steady flames, flickering flames, and Diesel engine exhausts. We present a novel LII technique for the determination of soot volume fraction by measuring the absolute incandescence intensity, avoiding the need for ex situ calibration that typically uses a source of particles with known soot volume fraction. The technique developed in this study further extends the capabilities of existing LII for making practical quantitative measurements of soot. The spectral sensitivity of the detection system is determined by calibrating with an extended source of known radiance, and this sensitivity is then used to interpret the measured LII signals. Although it requires knowledge of the soot temperature, either from a numerical model of soot particle heating or experimentally determined by detecting LII signals at two different wavelengths, this technique offers a calibration-independent procedure for measuring soot volume fraction. Application of this technique to soot concentration measurements is demonstrated in a laminar diffusion flame. PMID- 16270567 TI - Frequency-resolved absorption tomography with tunable diode lasers. AB - A tunable diode laser was used for absorption tomography in an axisymmetric atmospheric pressure flat-flame burner. A rapid tomographic inversion algorithm was used to facilitate the many reconstructions at a relatively sparse set of projections typical of laser absorption tomography. Profiles of temperature and CO2 mole fraction were measured simultaneously in methane-air flames. Absorption measurements were made near the R-branch bandhead at 4.17 microm to minimize interferences with other species, while providing good temperature and concentration sensitivity at flame conditions. The procedure showed the advantage of reconstructing detailed spectra at each radial node. PMID- 16270569 TI - Efficient oxidizing agents for determination of 2,10-disubstituted phenothiazines. AB - 2,10-Disubstituted phenothiazines are the best drugs in psychiatry. Several methods for their analysis have been reported in the literature. The official methods are based on non-aqueous titration or spectrophotometry. Various oxidizing agents have been used for the spectrophotometric determination of 2,10 disubstituted phenothiazines, e.g. Ce(SO4)2, NH4VO3, K2S208, KIO4, KIO3, KBrO3, FeCl3, NaNO2, H2O2, chloramine T, p-benzoquinone, N-bromosuccinimide. Oxidation reactions of phenothiazines were also used for their determination by flow injection methods. PMID- 16270568 TI - Rapid determination of decabromodiphenyl ether in polystyrene by thermal desorption-GC/MS. AB - A rapid determination of decabromodiphenyl ether (DeBDE) in polystyrene (PS) by thermal desorption (TD)-GC/MS was studied. The TD-GC/MS method using a pyrolysis GC/MS system allowed the quick quantification of DeBDE in a waste TV back plate composing of a PS flame-retarded with polybrominated diphenyl ethers on the basis of the resulting chromatogram with a ca. 4% relative standard deviation without using any tedious sample pretreatment, such as solvent extraction. PMID- 16270570 TI - Hydrodynamic electrochemistry in 20 microL drops in the rotating sample system. AB - The Rotating Sample System (RSS) has been conceived in the authors' laboratory as a convection platform for microliter-sized solution volumes. Convection is achieved by rotating a small drop of sample on a stationary substrate by humidified gas jets directed tangentially at the drop base with the working electrode and a liquid junction embedded in it. Simplicity and portability of the device, and substrates complete with microfabricated electrode and junction made potentially disposable, are further competitive advantages with respect to competing, conventional analytical systems. In this work the RSS' performance with variation of system parameters such as the position and size of gas jets used for sample rotation, and position of the working electrode in the substrate are studied. Trace levels of Pb could be detected with this system and is reported here. PMID- 16270571 TI - Determination of three bioactive constituents in moutan cortex by capillary electrophoresis with electrochemical detection. AB - Capillary electrophoresis with electrochemical detection has been employed for the separation and determination of the three active constituents (paeonol, benzoyloxypaeoniflorin, and oxypaeoniflorin) in traditional Chinese medicine, Moutan Cortex (root cortex of Paeonia suffruticosa Andrews). The effects of several important factors, such as the concentration of running buffer, the separation voltage, the injection time, and the detection potential, were investigated to determine the optimum conditions. The detection electrode was a 300 microm diameter carbon-disc electrode at a working potential of +0.90 V (versus SCE). The three analytes could be well separated within 7 min in a 40 cm length fused-silica capillary at a separation voltage of 12 kV in a 50 mM borate buffer (pH 9.2). The relation between the peak current and the analyte concentration was linear over 3 orders of magnitude with detection limits (S/N = 3) ranging from 0.4 to 0.7 microM for all analytes. The proposed method has been successfully applied to the determination of paeonol, benzoyloxypaeoniflorin, and oxypaeoniflorin in real plant samples with satisfactory assay results. PMID- 16270572 TI - Effect of the silanization conditions on chromatographic behavior of an open tubular capillary column coated with a modified silica-gel thin layer. AB - The performance of an open-tubular capillary column coated with a modified silica gel thin layer was investigated, particularly concerning the effect of the silanization process on it. Although the increase in the octadecyltriethoxysilane (ODTES) concentration in the silanization process could enhance the retention factor of naphthalene, its theoretical plate number was significantly reduced (ODTES, 5 to 50%; k, 0.2 to 4.3; N, 79600 to 2600 m(-1)). Namely, the increase in the retention factor was accompanied by a decrease in the theoretical plate number. A similar phenomenon was also observed when octadecyldimethylchlorosilane (ODCS) was used as the silanization regent. However, increases in both the retention factor and the theoretical plate number could be achieved (sample, naphthalene; k, 0.05 to 0.09; N, 149000 to 220000 m(-1)) by a NaOH treatment to the fabricated thin porous silica-gel layer before silanization with ODCS. The electrochromatographic separation of proteins and peptides by using the NaOH treated column could obtain more peaks than electrophoretic separation. PMID- 16270573 TI - Extraction capacity of diglycolamide derivatives for Ca(II), Nd(III) and Zr(IV) from nitric acid to N-dodecane containing a solvent modifier. AB - In order to evaluate the extraction property of new extractants, diglycolamide (DGA) compounds, we investigated the maximum extraction of di-, tri-, and tetravalent metal ions using nitric acid and n-dodecane. The limits of metal concentration (LOC) for Ca(II), Nd(III) and Zr(IV) in the organic phase are strongly influenced by HNO3 and the extractant concentration. For the purpose of enhancing the LOC value, we employed a modifier of the solvent, N,N-dihexyl octanamide (DHOA) and DGA with a long alkyl chain, and examined the results. It was evident that LOC increased with the DHOA concentration and the length of the alkyl chain attached to the N atom of DGA. The stoichiometric values of LOC(Zr) estimated from the extraction reaction were confirmed by using the extraction condition: tetraoctyl-DGA/1 M DHOA + n-dodecane and 3 M HNO3. PMID- 16270574 TI - Analysis of the monosaccharide components in Angelica polysaccharides by high performance liquid chromatography. AB - An analytical method of on-line high performance liquid chromatography (HLPC) was developed to simultaneously separate and identify the monosaccharide composition of three Angelica polysaccharide fractions (APF), named APF1, APF2 and APF3. In this method, APF were hydrolyzed into component monosaccharides and subsequently labeled with 1-phenyl-3-methyl-5-pyrazolone (PMP), and then the labeled monosaccharide derivatives were separated by a reverse-phase C18 column and monitored by UV absorbance at 250 nm. The results showed that nine monosaccharide derivatives have been well separated by HPLC under optimized conditions and the composition analysis of monosaccharides from APF samples could be achieved using acid hydrolysis and a set of monosaccharide standards. With this method, the within-day and day to day precisions of the composition determinations were 3.41 4.87% and 3.12-4.93% (RSD), respectively. The method was successfully applied to the determination of the component monosaccharides of Angelica polysaccharides. PMID- 16270575 TI - Comparison of the acid combinations in microwave-assisted digestion of marine sediments for heavy metal analyses. AB - Sediments as a tool for monitoring contamination by heavy metals in the environment has long been considered. It is therefore a necessity to produce reliable data for such purposes. Microwave-assisted acid dissolution has proved to be a suitable method for digesting complex matrices, such as sediments. However, due to the infancy of the technique, the procedures are numerous and varied in both the reagents used and microwave conditions. In this study, the efficiency of two recommended acid mixtures, a HNO3-HF mixture and an aqua regia HF mixture, under the same microwave digestion conditions were compared using certified reference materials. It was observed that the HNO3-HF mixture showed better efficiency than the aqua regia-HF mixture in most of the heavy metals analyzed in all certified reference materials used. PMID- 16270576 TI - Preconcentration of rare earth elements on silica gel loaded with 1-phenyl-3 methyl-4-benzoylpyrazol-5-one prior to their determination by ICP-AES. AB - A new chelating resin, silica gel loaded with 1-phenyl-3-methyl-4-benzoylpyrazol 5-one (PMBP), was prepared and used for the preconcentration of trace amounts of rare earth elements (REEs) in water samples prior to their determination by inductively coupled plasma atomic emission spectrometry (ICP-AES). REEs (La, Eu, Yb and Y) were quantitatively retained on the column packed with modified silica gel in the pH range 5 - 8 and separated from the matrix, and then recovered by eluting with 2.0 mol L(-1) HNO3. The adsorption capacity of modified silica gel for La, Eu, Yb and Y was 0.208, 0.249, 0.239 and 0.224 mmol g(-1), respectively. The method has been successfully applied for the determination of La, Eu, Yb and Y in geological and environmental samples with satisfactory results. PMID- 16270577 TI - Automation of liquid-liquid extraction-spectrophotometry using prolonged pseudo liquid drops and handheld CCD for speciation of Cr(VI) and Cr(III) in water samples. AB - A simple spectrophotometric system, based on a prolonged pseudo-liquid drop device as an optical cell and a handheld charge coupled device (CCD) as a detector, was constructed for automatic liquid-liquid extraction and spectrophotometric speciation of trace Cr(VI) and Cr(III) in water samples. A tungsten halogen lamp was used as the light source, and a laboratory-constructed T-tube with two open ends was used to form the prolonged pseudo-liquid drop inside the tube. In the medium of perchloric acid solution, Cr(VI) reacted with 1,5-diphenylcarbazide (DPC); the formed complex was automatically extracted into n-pentanol, with a preconcentration ratio of about 5. The organic phase with extracted chromium complex was then pumped through the optical cell for absorbance measurement at 548 nm. Under optimal conditions, the calibration curve was linear in the range of 7.5 - 350 microg L(-1), with a correlation coefficient of 0.9993. The limit of detection (3sigma) was 7.5 microg L(-1). That Cr(III) species cannot react with DPC, but can be oxidized to Cr(VI) prior to determination, is the basis of the speciation analysis. The proposed speciation analysis was sensitive, yet simple, labor-effective, and cost-effective. It has been preliminarily applied for the speciation of Cr(VI) and Cr(III) in spiked river and tap water samples. It can also be used for other automatic liquid liquid extraction-spectrophotometric determinations. PMID- 16270578 TI - An infrared evanescent wave sensing system coupled with a hollow fiber membrane for detection of volatile organic compounds in aqueous solutions. AB - We have developed an on-line sensing method for the detection of volatile organic compounds (VOCs) in contaminated aqueous solutions by combining a microporous hollow fiber membrane with an infrared (IR) sensing system. Polypropylene microporous hollow fibers were used to separate the VOCs from the aqueous solution into the hollow fibers, which were purged countercurrently for detection by the IR sensing systems. An evanescent-wave-type IR sensing system was used to detect the VOCs that were purged from the hollow fibers. The sensing element was coated with polyisobutylene (PIB) to concentrate the VOCs for their detection. To study the performance of this system, we examined a number of factors, such as the purging flow rate, the sample flow rate, and the volatilities of the VOCs. The results indicate that an increase in the purging flow rate reduces the analytical signal significantly, especially for purging flow rates >2 mL/min. The pumping flow rate for the aqueous sample also influenced the analytical signals, but far less sensitively. The volatilities of the examined compounds also affected the analytical signals: the higher the volatility of the compound, the lower the intensity of the analytical signals and the shorter the time required to reach the equilibrium signal. From an examination of the dynamic range of this proposed method, a regression coefficient >0.994 was obtained for concentrations below 250 mg/L, even under non-equilibrium conditions. The response time of the system was studied in an effort to examine the suitability of using this sensing method for automatic detection. The results indicate that new equilibrium conditions were established within 3 min for highly volatile compounds, which suggests that on-line monitoring of the levels of VOCs can be performed in the field. PMID- 16270579 TI - Resolution of 13 polycyclic aromatic hydrocarbons by constant-wavelength synchronous spectrofluorometry. AB - A method capable of determining 13 PAHs (acenaphthene, anthracene, benzo[a]anthracene, benzo[a]pyrene, benzo[b]fluoranthene, benzo[k]fluoranthene, chrysene, dibenzo[ah]anthracene, fluoranthene, fluorene, indene[1,2,3-cd]pyrene, phenanthrene and pyrene) in a mixture of 16 EPA PAHs by second derivative synchronous spectrofluorometry in the constant wavelength mode was developed. It has not been possible to determine the following PAHs in the mixture: acenaphthylene, benzo[ghi]perylene and naphthalene. The approach studied allows the sensitive, rapid and inexpensive identification and quantitation of 13 PAHs in a solution of hexane. The detection limits are <1 microg L(-1) (except for chrysene and phenanthrene). PMID- 16270580 TI - Determination of heparin using ciprofloxacin-Tb3+ as a fluorescence probe. AB - A new spectrofluorometric method was developed for the determination of trace amounts of heparin (Hep). Using ciprofloxacin (CIP)-terbium (Tb3+) as a fluorescent probe, in a buffer solution of pH 7.20, Hep can remarkably enhance the fluorescence intensity of the CIP-Tb3+ complex at lambda = 545 nm; also, the enhanced fluorescence intensity the Tb3+ ion is proportional to the concentration of Hep. The optimum conditions for the determination of Hep were also investigated. The dynamic range for the determination of Hep is 0.1 - 1.2 microg ml(-1) with a detection limit of 6.89 ng ml(-1). This method is simple, practical and relatively free of interference from coexisting substances, and can be successfully applied to assess Hep in biological samples. By the Rosenthanl graphic method, the association constant and binding numbers of heparin with the probe are 2.44 x 10(5) l mol(-1) and 19.7. Moreover, the enhancement mechanisms of the fluorescence intensity in the CIP-Tb3+ system and the CIP-Tb3+-Hep system have also been considered. PMID- 16270581 TI - Kinetic determination of thiocyanate on the basis of its catalytic effect on the oxidation of methylene blue with potassium bromate. AB - This study reports a sensitive kinetic spectrophotometric method for the determination of trace amounts of thiocyanate. In acidic solution, Methylene Blue (MB) is oxidized by bromate to form a colorless compound. The reaction is accelerated by trace amounts of thiocyanate and can be followed by measuring the absorbance at 664 nm. The absorbance of the reaction decreased with an increase in the reaction time. Under the optimum experimental conditions (0.56 M of sulfuric acid, 3.9 x 10(-5) M of MB, 3.0 x 10(-3) M of bromate, 180 s, 25 degrees C), thiocyanate can be determined in the range 5.0 - 180 ng/ml. The relative standard deviations (n = 8) are 2.81 and 1.43% for 10.0 and 150 ng/ml thiocyanate, respectively. The detection limit of this method is (3sigma) 3.8 ng/ml. This method was successfully applied to the determination of thiocyanate in real samples. PMID- 16270582 TI - Fabrication of herbicide biosensors based on the inhibition of enzyme activity that catalyzes the scavenging of hydrogen peroxide in a thylakoid membrane. AB - A novel herbicide biosensor with a thylakoid modified membrane electrode is presented. Thylakoid, isolated from spinach leaves, was entrapped in a membrane of poly (vinylalcohol) with the styrylpyridinium group (PVA-SbQ). The thylakoid membrane was fixed on the surface of a platinum electrode. It was found that the enzymes in thylakoid kept their activity for several months in the membrane. The oxidative current of hydrogen peroxide in a Tris-HCl buffer solution (pH 7.4) was detected at the modified electrode by a differential pulse voltammetric method. In the presence of herbicides, the oxidation current from the hydrogen peroxide decreased due to an inhibitor effect on the enzymes in thylakoid compared with that in the absence of the herbicides. The changes in the oxidation current at the electrode were proportional to the herbicide concentrations. The sensor could be used to detect herbicides in concentration ranges of 3 x 10(-9) - 1.5 x 10(-7) M for paraquat, 1 x 10(-8) - 3 x 10(-7) M for diuron, 4 x 10(-8) - 3 x 10(-6) M for prometryn, 5 x 10(-8) - 5 x 10(-6) M for atrazine and 1 x 10(-7) - 5 x 10(-6) M for ametryn, respectively. The enzyme activity on scavenging hydrogen peroxide in the modified PVA-SbQ membrane was examined. PMID- 16270583 TI - Electrochemical behavior and detection of plant hormone 6-benzyl adenine in acetate buffer at mercury electrode. AB - The electrochemical behavior of 6-benzyl adenine (6-BA) has been studied in 0.1 mol L(-1) HAc-NaAc solution (pH 3.8). Cyclic voltammetry, single-sweep polarography and direct current polarography were employed to clarify the mechanism of the electrode process; the kinetic parameters of the rate determining step were determined. Reduction of 6-BA involves two pH-dependent processes, corresponding to the overall irreversible reduction of the 1,6 and 3,2 N=C bonds. Each reduction stage consists a preprotonation of the nitrogen atom at the electroactive site and a rapid two-electron transfer. In the presence of 6 BA, the reduction potentials of some ions were shifted. Under the given conditions, 6-BA displays one irreversible reduction peak controlled by adsorption. Two linear response were observed in the range 2.0 x 10(-8) - 8.0 x 10(-7) mol L(-1) and the range 1.0 x 10(-6) - 8.0 x 10(-6) mol L(-1), with correlation coefficients of 0.9995 and 0.9998, respectively. The detection limit is 7.1 x 10(-9) mol L(-1). The method had been applied to the determination of 6 BA in bean sprout samples with satisfactory results. PMID- 16270585 TI - Ultraviolet laser photo-modulation voltammetry of tetraphenylborate at a liquid/liquid interface. AB - Photo-modulation voltammetry was applied to detecting the photolysis of tetraphenylborate (TPhB) at a water/1,2-dichloroethane (DCE) interface by using a He-Cd laser emitting a beam with a major 325-nm line and minor lines of shorter wavelengths. When the interface was irradiated from the water-phase side, a new wave appeared in the photomodulation voltammogram, suggesting that TPhB was photolyzed and the anionic product was transferred across the interface. The concentration dependence of the photocurrents was successfully explained by a theory based on the photolytic process at the interface. PMID- 16270584 TI - Electrochemistry of a carbon microfiber adsorbed by cobalt nanoparticles. AB - The adsorption of cobalt nanoparticles on a carbon microfiber surface has been electrochemicaly detected. The redox processes observed in an electrochemical cell filled with redistilled water and equipped with the carbon fiber microelectrode modified by cobalt nanoparticles have been compared to those observed in an aqueous solution of Co2+ cations. The movement of the adsorbed nanoparticles has been demonstrated by the feedback capacitance-potential method. PMID- 16270586 TI - Spectrophotometric determination of urinary protein with o-sulfophenylfluorone metal complex. AB - A simple and sensitive spectrophotometric method for the determination of human serum albumin (HSA) was established based on the ternary complex-formation reaction of HSA with o-sulfophenylfluorone (SPF) as a xanthene dye and metal ion (niobium(V) and bismuth(III)) in the presence of a dispersion agent. This new method enabled the determination of HSA in the range of 1 - 15 microg/ml HSA by measuring the difference of the absorbance at 530 nm between HSA-SPF-metal ion and SPF-metal ion solutions. In the determination of HSA, this method is about 2 times more sensitive than the Pyrogallol Red-molybdenum(VI) method (PR method), which accounts for more than 80% of the quantification methods for urinary protein assays in Japan. There was no significant difference between the results obtained by the present method and the PR method for human urine samples. The binding process between the SPF-metal complex and HSA was studied by determining the binding parameters and the thermodynamic parameters. PMID- 16270587 TI - Determination method for steroid 5alpha-reductase activity using liquid chromatography/atmospheric pressure chemical ionization-mass spectrometry. AB - A determination method for steroid 5alpha-reductase activity using liquid chromatography/atmospheric pressure chemical ionization-mass spectrometry (LC/APCI-MS) in the positive-ion mode has been developed. The rat prostatic enzyme source was used and the enzymatically formed 5alpha-dihydrotestosterone and 5a-androstane-3alpha,17beta-diol were determined by LC/APCI-MS using absolute calibration curve method. The sum of the formed products was used as a measurement of the enzyme activity. This method was applied to kinetic study of this enzyme and inhibitory experiments using Finasteride as a model inhibitor. PMID- 16270588 TI - Observation of 1,6-anhydro-beta-maltose and 1,6-anhydro-beta-D-glucopyranose complexed with rubidium by NMR spectroscopy and electrospray ionization mass spectrometry. AB - A complex of 1,6-anhydro-beta-maltose with rubidium and that of 1,6-anhydro-beta D-glucopyranose with rubidium were characterized using 87Rb NMR spectroscopy, diffusion-ordered NMR spectroscopy (DOSY) and electrospray ionization mass spectrometry (ESI-MS). Although subtle differences were observed in the 1H chemical shifts of 1,6-anhydro-beta-maltose in between the presence and absence of rubidium in deuterium oxide, measurements of the spin-lattice relaxation time (T1) of the 87Rb nucleus, the diffusion coefficients of 1,6-anhydro-beta-maltose using 1H DOSY and ESI-MS indicated the complex formation of 1,6-anhydro-beta maltose with rubidium. The complex formation with rubidium was also identified for 1,6-anhydro-beta-D-glucopyranose using NMR and ESI-MS techniques. PMID- 16270589 TI - Electrochemical adsorptive behavior of some fluoroquinolones at carbon paste electrode. AB - Cyclic voltammetry and differential pulse voltammetry were used to explore the adsorption behavior of three antibacterial agents at a carbon paste electrode. The drugs were accumulated on a carbon paste electrode, and a well-defined oxidation peak was obtained in acetate buffer (pH 5.0). The adsorptive stripping response was evaluated as a function of some variables such as the scan rate, pH and accumulation time. A simple, precise, inexpensive and sensitive voltammetric method has been developed for the determination of the cited drugs (Lomefloxacin (LFX), Sparfloxacin hydrochloride (SFX), and Gatifloxacin (GFX)). A linear calibration was obtained from 2 x 10(-7) M to 4 x 10(-5) M for LFX, 2 x 10(-7) M to 6 x 10(-5) M for SFX, and GFX. The limits of detection (LOD) were 4.2 x 10( 7), 7 x 10(-7) and 6.6 x 10(-7) M, while the limits of quantification (LOQ) were 1.4 x 10(-6), 2.3 x 10(-6) and 2.2 x 10(-6) M for LFX, SFX, and GFX, respectively. The R. S. D. of five measurements at the 1 x 10(-6) M level were 0.4, 0.5 and 0.3 for LFX, SFX and GFX, respectively. The method was applied to the determination of LFX, SFX and GFX in dilute urine samples and dosage forms, and compared with the HPLC method. PMID- 16270590 TI - The power of partnership--charting new courses for RNs. PMID- 16270591 TI - 2006 Registration Package will introduce members to HPA and the Continuing Competence Program. PMID- 16270592 TI - Healthy solution. Judgement in clinical practice--"it seemed like a good idea at the time". PMID- 16270593 TI - Surgical revision of the failed obliterated frontal sinus. AB - BACKGROUND: Surgical revision of failed frontal sinus obliteration, traditionally, has been limited to repeat obliteration. However, endoscopic techniques may be successful in selected cases. We review our experience in surgical revision of failed frontal obliteration and propose a management algorithm. METHODS: Retrospective chart review was performed over a 5-year period for patients who presented for surgical revision of a previously obliterated frontal sinus. Indications for surgery, radiological findings, and surgical approach were reviewed. RESULTS: Nineteen patients were identified, presenting an average of 9.7 years from the initial obliteration. Eighty-four percent (n = 16) were approached endoscopically and 16% (n = 3) were approached by revision obliteration. The mean follow-up was 25 months. In the endoscopic group, patients had either mucoceles in the inferomedial aspect of the frontal sinus or incomplete obliteration with persistent disease in the pneumatized frontal remnant. Eighty-one percent (13/16) were managed successfully with a single endoscopic procedure. Nineteen percent (3/16) had persistent disease requiring either a subsequent obliteration or Riedel ablation because of infected fat graft or frontal osteomyelitis. All patients who were managed successfully endoscopically remained free of disease with patent frontal sinusotomies throughout the follow-up period. The endoscopic failures required one to two additional external procedures to achieve disease resolution. In the revision obliteration group, all patients had mucoceles in either the lateral or the superior frontal sinus. All three patients had resolution of disease after a single procedure and remained free of disease throughout the follow-up period. CONCLUSION: Selected patients undergoing revision of frontal obliteration may benefit from endoscopic approaches. If disease is localized in the frontal recess or inferomedial frontal sinus, endoscopic management may be successful in the majority of patients. Superior or lateral frontal disease appears to be best approached externally. Patients undergoing endoscopic salvage should be counseled about the possible need for revision obliteration if disease persists. PMID- 16270594 TI - Image-guided osteoplastic frontal sinusotomy. AB - BACKGROUND: A 6-ft Caldwell radiograph template and transillumination, typically, are used to elucidate the frontal sinus outline for osteoplastic frontal sinusotomy (OFS). These techniques can be fraught with imprecision. The consequences of imprecise OFS may result in significant complications. Computer aided surgery may offer a safe and accurate alternative to these techniques in selected cases. Several disadvantages were noted with early computer-aided assistance. Current infrared and electromagnetic systems have eliminated many of these disadvantages. We describe our technique and experience with an infrared image-guidance system (The LandmarX Evolution; Medtronic Xomed, Jacksonville, FL) to create a precise OFS that maximizes exposure while minimizing morbidity. METHODS: We describe the use of an infrared image guidance system, the LandmarX Evolution for OFS in three cases. RESULTS: The LandmarX Evolution allowed for accurate placement of the OFS in each of the three cases and successful treatment of two symptomatic frontal sinus osteomas and a recurrent inverted papilloma. No complications were encountered. CONCLUSION: Image-guided OFS results in a confident and accurate entry into the frontal sinus. Image-guided OFS creation should be considered for select cases (i.e., complete opacification and altered anatomy) where performing an OFS by standard techniques may increase the complication rate. More experience with the technique and increased accessibility to image-guided equipment must be made possible before establishing this technique as a standard. PMID- 16270595 TI - Combined endoscopic trephination and endoscopic frontal sinusotomy for management of complex frontal sinus pathology. AB - BACKGROUND: The advances in endoscopic sinus surgery have revolutionized the management of frontal sinus disease. Despite the successes, the purely endoscopic approach has its limitations, especially in patients with alterations in anatomy caused by previous surgical intervention or complex frontal sinus pneumatization patterns. The purpose of this study was to evaluate the efficacy of combined endoscopic trephination and endoscopic frontal sinusotomy (the above and below approach) in the management of these difficult cases. METHODS: Chart review was performed on patients undergoing the combined approach from October 1999 to June 2004. Demographic data, symptomatology, comorbidity, previous surgery, and primary pathology were determined. Outcome was assessed based on subjective symptom relief and objective endoscopic patency. RESULTS: Twenty-two patients with a mean age of 49.2 years underwent the combined approach. The primary pathology included mucoceles (15 patients), frontal sinusitis (2 patients), inverted papilloma (2 patients), osteoma (1 patient), fibrous dysplasia (1 patient), and pneumocephalus (1 patient). A total of 25 above and below procedures (22 primary and 3 revision procedures) were performed to manage the pathology. Postoperatively, headaches resolved in 47%, improved in 35%, and remained unchanged in 18% of the patients. Orbital symptoms resolved in 63%, improved in 25%, and remained unchanged in 12% of the patients. Endoscopic patency of the frontal sinusotomy was confirmed in 19 of 22 cases (86%) at a mean follow-up of 16.2 months. CONCLUSION: Management of complex frontal sinus pathology may require adjunct approaches in conjunction to the standard endoscopic techniques. In this series, the above and below approach was used successfully in 22 patients. The combined approach may serve as an important adjunct for management of complex frontal sinus disease. PMID- 16270596 TI - Staged endoscopic and combined open/endoscopic approach in the management of inverted papilloma of the frontal sinus. AB - BACKGROUND: The endoscopic management of inverted papilloma has gained increasing popularity over the last 10 years. Although early concerns over an increased risk of recurrence seem to have been allayed, the appropriate management of lesions involving the frontal sinus and frontal recess still has to be determined. METHODS: We performed a retrospective review of the results of all patients with inverted papilloma from 2000 to 2004. RESULTS: A total of 18 patients were treated between October 2000 and January 2004. Six patients had frontal sinus involvement at the time of initial evaluation. One of these patients had isolated frontal sinus involvement. These patients were managed with either initial endoscopic resection with determination for the need for an additional procedure at the time of endoscopic resection (n = 5) or open/endoscopic approach for isolated frontal sinus involvement (n = 1). Of the five patients who had their disease managed endoscopically, three patients were determined at the initial procedure to need an osteoplastic flap and, subsequently, were managed successfully with a combined approach. One other patient was initially successfully managed endoscopically but ultimately required an osteoplastic flap for definitive management. The fifth patient was managed entirely endoscopically with multiple procedures. All patients treated with this protocol remain disease free with an average follow-up of 13.3 months. CONCLUSION: The limitations of endoscopic resection of inverted papilloma of the frontal recess can be managed with staged procedures. Initial endoscopic resection of ethmoid/maxillary disease with subsequent open treatment of the frontal sinus has been successful in our experience. PMID- 16270597 TI - Endoscopic management of extensive inverted papilloma. AB - BACKGROUND: Given the malignant potential and propensity for recurrence of inverted papilloma (IP) of the sinonasal cavity, complete excision is warranted. For disease extending to multiple sites, open surgical oncological procedures are associated with high morbidity and do not assure complete control of the tumor. The endoscopic approach provides excellent visualization, permits removal of diseased mucosa while preserving vital anatomic structures, and allows for excellent postoperative surveillance. Recurrences are identified early and endoscopic resection is repeated as necessary until there is no evidence of disease. METHODS: Data were prospectively collected and subsequently reviewed on 18 consecutive patients who underwent endoscopic management of extensive IP (present at more than one anatomic site) between 1999 and 2003. RESULTS: Fourteen men and four women with a mean age of 54 years (range, 36-74 years) were followed for an average of 29 months (range, 6-46 months) after initial endoscopic resection. Seventy-eight percent (14 patients) complained of nasal airway obstruction for more than 6 months and 22% (4 patients) were incidentally noted to have a nasal mass on endoscopy or computed tomography. Eleven patients had undergone therapeutic procedures on initial evaluation. The most common sites affected were maxillary sinus, lamina papyracea, and ethmoid sinus. Patients required an average of 1.6 endoscopic surgeries (range, 1-3 surgeries) to achieve local control; 10 patients (56%) required only one. All patients were symptomatically improved and complications were limited to one cerebrospinal fluid leak, which was repaired intraoperatively. CONCLUSION: Extensive IP can be controlled using minimally invasive endoscopic procedures as long as close follow up is maintained. Operative risk and postoperative morbidity are significantly less than observed with open procedures. PMID- 16270598 TI - Demonstration of biofilm in human bacterial chronic rhinosinusitis. AB - BACKGROUND: Bacterial biofilms may explain why some patients with bacterial chronic rhinosinusitis (CRS) improve while on antibiotics but relapse after completion of the antibiotic. In the human host, biofilms exist as a community of bacteria surrounded by a glycocalyx that is adherent to a foreign body or a mucosal surface with impaired host defense. Biofilms generate planktonic, nonadherent bacterial forms that may metastasize infection and generate systemic illness. These planktonic bacteria are susceptible to antibiotics, unlike the adherent biofilm. METHODS: We reviewed four cases of CRS using transmission electron microscopy (TEM) to assay for typical colony architecture of biofilms. Bacterial communities surrounded by a glycocalyx of inert cellular membrane materials consistent with a biofilm were shown in two patients. RESULTS: In the two patients without biofilm, a nonbacterial etiology was discovered (allergic fungal sinusitis) in one and in the other there was scant anaerobic growth on culture and the Gram stain was negative. Culture of the material from the biofilm grew Pseudomonas aeruginosa in both patients. Pseudomonas from the biofilm showed a glycocalyx, not present in Pseudomonas cultured for 72 hours on culture media. Both patients' symptoms with bacterial biofilms were refractory to culture directed antibiotics, topical steroids, and nasal lavages. Surgery resulted in cure or significant improvement. CONCLUSION: Biofilms are refractory to antibiotics and often only cured by mechanical debridement. We believe this is the first TEM documentation of bacterial biofilms in CRS in humans. PMID- 16270599 TI - Bacterial location in chronic sinusitis. AB - BACKGROUND: In chronic nonallergic sinusitis, it is often assumed that bacteria invade the sinus mucosa where the inflammatory condition begins and is maintained. However, the bacterial presence in a normal or moderately damaged epithelial layer has never been proved in biopsy studies. METHODS: In this study, mucosal samples from six consecutive patients with chronic sinusitis were examined. Transmission electron microscopy was used and the presence of bacterial invasion and formation of phagosomes containing bacteria as a marker of host response were studied. RESULTS: Phagocytosis of bacteria was observed in the sinus mucosa in samples from only one patient. In the other five patients, no signs of phagocytosis were seen. CONCLUSION: Based on these results, we concluded that in chronic sinusitis, bacterial invasion in sinus mucosa is not an obligatory phenomenon. PMID- 16270600 TI - Concordance of middle meatal swab and maxillary sinus aspirate in acute and chronic sinusitis: a meta-analysis. AB - BACKGROUND: The medical management of acute and chronic sinusitis is a therapeutic challenge. The use of endoscopic middle meatal cultures as a noninvasive method to determine the bacteriology of the maxillary sinus has not accurately been established. The aim of this study was to review the literature that compares cultures obtained by endoscopic middle meatal swabs with those obtained from maxillary sinus aspirates (MSAs). METHODS: We reviewed studies published between January 1966 and October 2003 that were identified from searches of multiple databases, bibliographies, and original articles. Studies were included for analysis if they compared the results of endoscopic middle meatal cultures to aspirate cultures. All clinical variables and test performances were independently extracted by two reviewers. RESULTS: Middle meatal culture had a per isolate accuracy of 82% (95% confidence interval, 0.64, 0.92) compared with MSA (excluding coagulase negative staphylococcus and fungal cultures). CONCLUSION: Endoscopic middle meatal cultures have a high concordance with MSAs. PMID- 16270601 TI - Evaluation of postoperative pain after sinonasal surgery. AB - BACKGROUND: Pain after sinonasal surgery concerns both patients and surgeons. Factors affecting sinonasal postoperative pain have not been examined extensively. METHODS: Using a prospective survey design, sinonasal surgery patients evaluated postoperative pain (on a scale of 0-10), pain location, and medication use. RESULTS: One hundred twenty-seven patients consented to participate. One hundred fifteen patients returned surveys, with 100 patients appropriate for analysis over the 6-day postoperative period. Pain score and medication use were evaluated with respect to sex, primary/revision case, nasal packing, and other factors. Repeated measures analysis of variance and chi-square analysis were conducted (p < 0.05). Pain ratings and analgesic use declined significantly over the postoperative period (p < 0.05). Mean pain score on postoperative day (POD) 1 was 3.61 and on POD 6 was 1.72. Mean medication use was 1.37 tablets on POD 1 and 0.55 tablets on POD 6. Additionally, a significant interaction existed, such that narcotic medication use declined from 1.91 tablets on POD 1 to 0.52 tablets on POD 6, whereas nonnarcotic medication use remained steady (p < 0.05). Periorbital pain was most frequent (46.3%), with unilateral facial pain reported least (4.1%). A significant difference existed for distribution of pain location (p < 0.05). Finally, the difference in pain rating between primary and revision procedure for women (0.65) was less than for men (1.12); this interaction was significant (p < 0.05). CONCLUSION: Multiple patient and operative factors affect pain rating and medication use after sinonasal surgery. Generally, pain level should be low with little analgesic use postoperatively. PMID- 16270602 TI - Surgical revision of the post-Caldwell-Luc maxillary sinus. AB - BACKGROUND: Endoscopic sinus surgeons are commonly faced with the management of patients with persistent maxillary sinusitis despite previous Caldwell-Luc surgery. Given the potential for altered mucociliary clearance in the post Caldwell-Luc maxillary sinus, the optimal approach for surgical revision has not been well characterized. The objective of this study was to review our experience in endoscopic versus repeat Caldwell-Luc approaches in patients who have failed Caldwell-Luc surgery for chronic maxillary sinusitis. METHODS: Retrospective chart review was performed on patients who had a history of a Caldwell-Luc procedure and who then underwent a surgical revision for persistent maxillary sinusitis at the Oregon Health and Science University and Medical College of Wisconsin between 1983 and 2002. RESULTS: Sixty-eight patients were identified, with a total of 156 revision procedures performed on 103 maxillary sinuses. Fifty three percent of the sinuses underwent endoscopic maxillary antrostomy as the first surgical revision, while 47% underwent a revision Caldwell-Luc procedure. Sixty-seven percent of the sinuses in the revision endoscopic group had clinical resolution with a single surgical revision, and 60% of the sinuses in the revision Caldwell-Luc group had clinical improvement with one surgical revision (p = 0.46). The endoscopic group averaged 1.3+/-0.5 revision procedures per sinus to achieve clinical resolution, and the revision Caldwell-Luc group averaged 1.7+/-1.0 revision procedures per sinus (p = 0.3). Mean follow-up was 25 months. CONCLUSION: Endoscopic revision of the maxillary sinus yields comparable outcomes to repeat Caldwell-Luc procedure in patients with a history of previous failed Caldwell-Luc surgery. Endoscopic revision surgery is a viable alternative for surgical rehabilitation of the post-Caldwell-Luc maxillary sinus. PMID- 16270603 TI - Annexin A1 expression in nasopharyngeal carcinoma correlates with squamous differentiation. AB - BACKGROUND: Alterations of annexin A1 (ANXA1) expression have been reported in various cancers. However, no data are available about the expression of this protein in nasopharyngeal carcinomas (NPCs). The objective of this study was to investigate the expression of ANXA1 in these tumors. METHODS: We examined noncancerous nasopharyngeal mucosa (4 cases) and NPC (20 cases) for ANXA1 expression using immunohistochemistry. RESULTS: All tumor tissues showed markedly reduced ANXA1 expression compared with a strong positive signal observed in the corresponding normal epithelia. We found that ANXA1 expression is associated with the histological type in NPC. Only squamous cell carcinomas presented a positive ANXA1 signal in differentiated areas whereas all poorly differentiated tumors exhibited negative staining. CONCLUSION: Our data show for the first time that ANXA1 expression is down-regulated in NPC and that its expression seems to be related with the squamous differentiation status of these tumors. PMID- 16270604 TI - Bipolar radiofrequency plasma-mediated ablation of porcine nasal septal cartilage: a pilot investigation. AB - BACKGROUND: The objectives of this pilot study were to (1) determine whether bipolar radiofrequency plasma-mediated ablation (PMA) can efficiently remove nasal septal cartilage and (2) calculate the ablation rate as a function of device power, probe force, and translation velocity using ex vivo porcine tissue. METHODS: Specimens were secured to a linear translation stage and were subjected to varying translation velocities (4, 7, and 12 cm/s), probe forces (140, 200, and 225 g), and bipolar radiofrequency generator (Coblator ENTec power settings, 38-58, 77-115, and 129-193 Watts root mean squared. Specimen mass loss and depth of ablation were measured using an analytic balance and dissection microscope, respectively. RESULTS: Visual and microscopic inspection revealed little char. Mass loss increased with decreasing translation velocity and increasing generator setting. Increasing probe force also influenced mass loss and increased crater depth. Depth of ablation did not correlate with translation velocity or generator setting. CONCLUSION: PMA effectively ablates nasal septal cartilage and may be able to reduce or contour cartilaginous deformities and framework structures in the head and neck. PMID- 16270605 TI - Pharmacological characterization of postjunctional alpha-adrenoceptors in human nasal mucosa. AB - BACKGROUND: Functional alpha1- and alpha2-adrenoreceptor subtype pharmacology was characterized in an in vitro human nasal mucosa contractile bioassay. METHODS: Nasal mucosa was obtained from 49 donor patients and mucosal strips were placed in chambers filled with Krebs-Ringer solution and attached to isometric force transducers. RESULTS: Nonselective a-adrenoreceptor agonists epinephrine, norepinephrine, and oxymetazoline produced concentration-dependent contractions of isolated human nasal mucosa (pD2 = 5.2, 4.9, and 6.5, respectively). The alpha2-adrenoreceptor agonist BHT-920 (10 microM)-induced contractions were blocked by yohimbine (0.01-1 microM) and prazosin (0.01-1 microM) inhibited the contractile response to the alpha1-adrenoreceptor agonist phenylephrine (10 microM). Histological analysis showed that phenylephrine and BHT-920 differentially contracted the arteries and veins of human nasal mucosa, respectively. CONCLUSION: Our results indicate that functional alpha1- and alpha2 adrenoceptors are present and functional in human nasal mucosa. The alpha2 adrenoceptors display a predominant role in contracting the veins and the alpha1 adrenoceptors appear to preferentially constrict the human nasal arteries. PMID- 16270606 TI - Analysis of possible cross-contamination with the Venturi system atomizer. AB - BACKGROUND: The physics behind the Venturi atomizers suggest a possibility of bacterial colonization and the potential for patient cross-contamination. A protocol for use of the atomizer and clinically appropriate demonstration of cross contamination has not been established. METHODS: Three sterilized atomizers filled with a solution of 2% Pontocaine and 1% ephedrine (two test and one control) were used during a 5-day study period. Clinic staff was instructed to (1) use a nozzle tip, (2) use a nasal speculum, (3) avoid contact between the atomizer and the patient, and (4) apply a continuous, <1-second spray to the nasal cavity. Samples were obtained from each of the atomizers three times per day and plated on chocolate agar plates. The number and type of bacterial colony were registered. RESULTS: No respiratory pathogens grew from any of the 45 plated samples. Nine of 30 experimental atomizer samples produced 12 bacterial colonies. Of the 12 bacterial colonies obtained, 6 colonies of coagulase-negative Staphylococcus, 5 colonies of Corynebacterium sp., and 1 colony of Bacillus sp. were identified. One of 15 plated control atomizer samples produced two colonies of coagulase-negative Staphylococcus. An average of 1.3 colonies per positive test plate and an average of 2.0 colonies per positive control plate were identified. There was no evidence of an increasing number of colonies per plate or persistence of any particular bacteria identified over time to suggest contamination or colonization of the system reservoir. CONCLUSION: There is no risk of cross-contamination of patients with the use of the Venturi system atomizer as outlined in this study. Culture results from this study were consistent with random culture contamination during the plating and/or culturing period. There was no evidence to support the idea of bacterial colonization of the atomizers. Continued use of the Venturi system atomizer is an acceptable practice. PMID- 16270607 TI - Clinical strategy in hereditary hemorrhagic telangiectasia. AB - BACKGROUND: Hereditary hemorrhagic telangiectasia (HHT) is a recurrent bleeding tendency caused by vascular malformations and preferentially involving the mucous membrane of the nose. The rhinological management of epistaxis is a challenge in which the frequency of bleeding has to be reduced without damage to the nasal mucosa, despite the fact that therapy necessarily has to be repeated. METHODS: The clinical course in 30 patients with HHT was monitored prospectively. Nasal mucosal efflorescences underwent Nd:YAG laser therapy at individually defined intervals, and the effect on the frequency and duration of bleeding was documented, as were adverse effects. RESULTS: No serious adverse effects (e.g., septal defects or synechiae) were observed as a consequence of therapy. During the course of laser therapy and ongoing compliance with nasal mucosal care instructions, the frequency of bleeding fell from "several times daily" to "every 2 weeks." CONCLUSION: In conjunction with Nd:YAG laser therapy, ongoing and consistent care of the nasal mucosa is a proven and effective treatment regimen in HHT. As an integral element in an interdisciplinary strategy for diagnosis and therapy, this regimen yields satisfactory quality of life while avoiding local complications. PMID- 16270608 TI - The effect of the total intravenous anesthesia compared with inhalational anesthesia on the surgical field during endoscopic sinus surgery. AB - BACKGROUND: Bleeding during endoscopic sinus surgery (ESS) may increase complications and negatively effect the surgery and its outcome. The aim of this study was to compare the surgical field in patients in whom total intravenous anesthesia (TIVA) is used as opposed to inhalation anesthesia. A prospective randomized controlled trial was performed. METHODS: Fifty-six patients undergoing ESS were randomly assigned to receive either inhaled sevoflurane with incremental doses of fentanyl (n = 28) or TIVA via a propofol and remifentanil infusion (n = 28) for their general anesthesia. The surgical field was graded every 15 minutes using a validated scoring system. RESULTS: The two groups were matched for surgical procedure and computed tomography scores. Patients in the TIVA group were found to have a significantly lower surgical grade score than in the sevoflurane group (p < 0.001). Surgical grade score increased with time in both groups. Mean arterial pressure and pulse were found to influence the surgical field independently (p = 0.003 and p = 0.036 respectively). Mean surgical field grade scores were higher in the patients with allergic fungal sinusitis and nasal polyposis as opposed to chronic rhinosinusitis without polyps or fungus. Lund Mackay computed tomography scores were found to correlate positively with surgical grade (Spearman rank correlation, p = 0.001). CONCLUSION: In patients undergoing ESS, TIVA results in a better surgical field than inhalational anesthesia. PMID- 16270609 TI - Resection of anterior skull base tumors: comparison of combined traditional and endoscopic techniques. AB - BACKGROUND: Traditional craniofacial resection (tCFR) has been used successfully for resection of anterior skull base (ASB) tumors. Minimally invasive endoscopic resection (MIER) also has been used recently; this strategy facilitates superior visualization, avoids facial incisions, and preserves local structures. The goal of this study was to compare the outcome for these two approaches. METHODS: Retrospective chart analysis was conducted to identify patients undergoing resection of ASB tumors between January 1995 and January 2003. Demographic data, tumor characteristics, and the surgical approach used were determined. The mean operative time, estimated blood loss, hospital stay, and complications were analyzed. Recurrence and mortality rates were calculated. RESULTS: Nine patients were managed with the MIER approach, and 16 patients were treated with the traditional open approach. No significant difference was observed between groups on operative time, estimated blood loss, or hospital stay. Major complications were encountered in 2/9 (22%) and 7/16 (44%) patients in the MIER and tCFR groups, respectively. Recurrence was observed in 3/9 (33%) and 5/14 (36%) of the patients in the MIER and tCFR groups, respectively. Mortality rates in the MIER and tCFR groups were 0/9 (0%) and 4/15 (27%), respectively. CONCLUSION: In this preliminary study, MIER of ASB neoplasia did not differ significantly from tCFR in operative time, estimated blood loss, hospital stay, or complication rate. Survival and recurrence rates were similar also. This early experience suggests that MIER is a viable alternative for the surgical management of ASB lesions in appropriately selected patients. PMID- 16270611 TI - Literature and today's physician. PMID- 16270610 TI - Objective versus subjective outcomes assessment in rhinology. AB - BACKGROUND: Prior studies on the relationship between computed tomography scan findings and patient-based quality of life in chronic rhinosinusitis (CRS) have found very low correlations. Whereas surprising, similarfindings have been noted in other diseases. METHODS: We performed a cross-sectional analysis of the association between objective and subjective findings in nasal septal deformity and a systematic review and synthesis of the literature on CRS and other diseases. RESULTS: We found no association between objective anatomic findings and subjective symptoms in nasal obstruction (R = 0.03; Kruskal-Wallis test, p = 0.97). Multiple studies in CRS and other diseases-sleep apnea, hearing loss, asthma, etc.- have found similarly low correlations between objective and subjective testing. CONCLUSION: For nasal septal deviation and CRS, the patient's subjective perception of disease severity has, at best, a very weak association with objective assessment of severity. Patient-based outcomes assessment remains important; these instruments apparently quantify an aspect of disease not detected by objective testing. PMID- 16270612 TI - Drug samples--there is no free lunch! Minimize your risks and meet your regulatory responsibilities. PMID- 16270613 TI - Medicaid reform to impact our State's most vulnerable populations. PMID- 16270614 TI - The implications of rising medical student debt. PMID- 16270616 TI - Overweight children: what doctors can do to help. PMID- 16270617 TI - Vascular smooth muscle cell microparticles and coronary no-reflow: another piece of the puzzle? PMID- 16270618 TI - Regenerative cardiology: there are various ways to prosper. PMID- 16270619 TI - Stem cells for clinical use in cardiovascular medicine: current limitations and future perspectives. AB - Cell transplantation is currently gaining a growing interest as a potential new means of improving the prognosis of patients with cardiac failure. The basic assumption is that left ventricular dysfunction is largely due to the loss of a critical number of cardiomyocytes and that it can be partly reversed by implantation of new contractile cells into the postinfarction scars. Primarily for practical reasons, autologous skeletal myoblasts have been the first to undergo clinical trials and now that the feasibility of the procedure is well established, efficacy data are expected from the ongoing randomized studies. Bone marrow stem cells are also generating a great deal of interest, particularly in patients with acute myocardial infarction, and are currently undergoing extensive clinical testing although recent data have raised a cautionary note about the transdifferentiation potential of these cells. While experimental studies and early-phase clinical trials tend to support the concept that cell therapy may enhance cardiac repair, several key issues still need to be addressed including (1) the optimal type of donor cells in relation to the clinical profile of the patients, (2) the mechanism by which cell engraftment improves cardiac function, (3) the optimization of cell survival, (4) the development of less invasive cell delivery techniques and (5) the potential benefits of cell transplantation in nonischemic heart failure. Current evidence suggests, however, that adult stem cells (myogenic or marrow-derived) fail to electromechanically integrate within the recipient heart, thereby mandating the search for second generation cell types able to achieve this goal which is the prerequisite for an effective enhancement of contractile function. Preliminary data suggest that cells that feature a true cardiomyogenic phenotype such as cardiac stem cells and cardiac precommitted embryonic stem cells may fall in this category and carry the potential for ensuring a true regeneration of dead myocardium. PMID- 16270620 TI - Circulating endothelial cells in atrial fibrillation with and without acute cardiovascular disease. AB - Normal adults have very few circulating endothelial cells (CECs) in their blood, but increased levels have been shown in association with conditions associated with endothelial damage such as myocardial infarction and stroke. As atrial fibrillation (AF) is associated with a hypercoagulable state and abnormalities of plasma indices of endothelial damage/dysfunction, we hypothesised that CECs would also be raised in this condition, and would correlate with these plasma markers. We measured CECs (by immunofluoresence) as an indicator of frank endothelial damage, alongside 3 plasma indices of endothelial perturbation: von Willebrand factor (vWf), soluble E-selectin and soluble thrombomodulin (sTM) (all ELISA) in 28 patients with chronic 'stable' AF, 63 patients with AF plus an acute cardiovascular or cerebrovascular event as positive controls, and 20 healthy subjects in sinus rhythm as negative controls. Chronic 'stable' AF patients had significantly higher levels of plasma vWf (p<0.001 ), but comparable numbers of CECs (p=0.1638) in comparison to healthy controls. In patients with AF associated with an acute cardiovascular or cerebrovascular event, levels of CECs (p<0.0001) and sTM (p=0.004), but not vWf or sEsel, were significantly increased in comparison to chronic 'stable' AF patients. Patients with uncomplicated AF have abnormal systemic endothelial damage/dysfunction, as evident by increased plasma vWf levels, but normal numbers of CECs, compared to subjects in sinus rhythm. However, following clinical complications, such as stroke or significant haemodynamic compromise, further endothelial disturbance (as indicated by high levels of sTM and CECs) suggests additional endothelial damage. PMID- 16270621 TI - Circulating endothelial cells in acute ischaemic stroke. AB - Increased numbers of CD146-bearing circulating endothelial cells (CECs) in the peripheral blood probably represent the most direct evidence of endothelial cell damage. As acute ischaemic strokes are associated with endothelial abnormalities, we hypothesised that these CECs are raised in acute stroke, and that they would correlate with the other indices of endothelial perturbation, i.e. plasma von Willebrand factor (vWf) and soluble E-selectin. We studied 29 hypertensive patients (19 male; mean age 63 years) who presented with an acute stroke and compared them with 30 high risk hypertensive patients (21 male; mean age 62 years) and 30 normotensive controls (16 male; mean age 58 years). CECs were estimated by CD146 immunobead capture, vWf and soluble E-selectin by ELISA. Patients with an acute ischaemic stroke had significantly higher numbers of CECs/ml of blood (p<0.001) plasma vWf (p=0.008) soluble E-selectin (p=0.002) and higher systolic blood pressure (SBP) as compared to the other groups. The number of CECs significantly correlated with soluble E-selectin (r=0.432, p<0.001) and vWf (r=0.349, p=0.001) but not with SBP (r=0.198, p=0.069). However, in multivariate analysis, only disease group (i.e. health, hypertension or stroke) was associated with increased CECs. Acute ischaemic stroke is associated with increased numbers of CECs. The latter correlate well with established plasma markers of endothelial dysfunction or damage, thus unequivocally confirming severe vasculopathy in this condition. However, the greatest influence on CECs numbers was clinical group. PMID- 16270622 TI - Bone morphogenetic proteins and vascular differentiation: BMPing up vasculogenesis. AB - Vasculogenesis is an important mechanism of blood vessel formation not only in embryos but also in adults. It may contribute to reparative effects of progenitor cell therapy in ischemic diseases such as myocardial infarction. Signaling pathways involved in embryonic development, including the BMP pathway, are reactivated in adult vasculogenesis. As a consequence knowledge about embryonic signaling events will help to understand blood vessel formation in the adult. The role of BMPs in embryonic development has been studied extensively in the past decades but only recently their role in vasculogenesis has been recognized. Gain and loss of function models indicate that BMPs stimulate vasculogenesis in the embryo as well as in the adult. Additionally, BMPs interact with other pathways involved in blood vessel formation, such asVEGF signaling. Studying novel molecules such as BMPER that modulate BMP activity and that are expressed in vascular cells will help to understand vasculogenetic signaling and may open up new therapeutic avenues in vascular disease. PMID- 16270623 TI - An optimized embryonic stem cell model for consistent gene expression and developmental studies: a fundamental study. AB - In vitro differentiation of embryonic stem (ES) cells results in generation of tissue-specific somatic cells and may represent a powerful tool for general understanding of cellular differentiation and development in vivo. Culturing of most ES cell lines requires murine embryonic fibroblasts (MEF), which may influence adventitiously the genetic differentiation program of ES cells. We compared the expression profile of key developmental genes in the MEF-independent CGR8 ES cell line and in the MEF-dependent D3 ES cell line. Using neomycin resistant MEFs we demonstrated that MEFs are able to contaminate the D3 ES cells even after removing the MEFs. Subsequently, optimal differentiation conditions were established for the differentiation of CGR8 ES cells into various germ layer cells. Detailed gene expression studies in differentiating CGR8 cells were done by RT-PCR analysis and by microarray analysis demonstrating a general trend of the assessed genes to be expressed either in 3 days- or 10-days old embryoid bodies (EBs) when compared to undifferentiated ES cells. Subsets within the various functional gene classes were defined that are specifically up- or down regulated in concert. Interestingly, the present results demonstrate that developmental processes toward germ layer formation are irreversible and mostly independent of the culture conditions. Notably, apoptotic and mitochondrial ribosomal genes were down- and up-regulated in 10-days old EBs, respectively, whereas compared to the 3-days old EBs whereas the activity of the extracellular signal-regulated kinase (ERK) 1/2 decreased with progressive development. This article defines a platform for ES cell differentiation and gene expression studies. PMID- 16270624 TI - Molecular and phenotypic analyses of human embryonic stem cell-derived cardiomyocytes: opportunities and challenges for clinical translation. AB - Differentiation of human embryonic stem cells (hESCs) into cardiomyocytes in culture may offer unique opportunities for modeling genetic disorders, screening potentially cardiotoxic pharmaceutical agents or replacing cells of the diseased heart. However, before clinical utility can be realized, numerous hurdles must be overcome. Comprehensive molecular and phenotypic characterization is required but has so far been restricted to cardiomyocytes derived from a limited subset of hESC lines. Thus, we have initiated analysis of cardiomyocyte differentiation and function from a further two independently derived lines, BG01 and HUES-7. The challenge of improving cardiac cell induction, enrichment and maturation must also be addressed to meet the demands of high throughput pharmaceutical screening or to provide sufficient cells to repair an infarcted heart. Transplanted cells must functionally integrate without inducing arrhythmias, while survival and evasion of immune surveillance must be accomplished without tumorigenicity. This review evaluates the opportunities presented by hESC-derived cardiomyocytes and the progress towards surmounting the challenges of clinical translation. PMID- 16270625 TI - In search of the best candidate for regeneration of ischemic tissues: are embryonic/fetal stem cells more advantageous than adult counterparts? AB - Human stem cells and progenitor cells from the bone marrow have been proposed for the regeneration of ischemic cardiac tissues. Early clinical trials indicate that infusion of autologous bone-marrow cells into the infarcted heart enhances ventricular function, albeit the long-term benefit remains to be ascertained. Alternatively, angiogenic growth factors could be used to stimulate the recruitment of vascular progenitor cells into tissues in need of regeneration. Unfortunately, in atherosclerotic patients, the curative potential of autologous stem cells might be impoverished by underlying disease and associated risk factors. Thus, research is focusing on the use of embryonic stem cells which are capable of unlimited self-renewal and have the potential to give rise to all tissue types in the body. Ethical problems and technical hurdles may limit the immediate application of embryonic stem cells. In the meanwhile, fetal hematopoietic stem cells,which have been routinely used to reconstitute the hematopoietic system in man, could represent an alternative, owing to their juvenile phenotype and ability to differentiate into vascular endothelial, muscular, and neuronal cell lineages. With progresses in stem cell expansion, the blood of a single cord could be sufficient to transplant an adult. These observations raise the exciting possibility of using fetal cells as a new way to speed up the healing of damaged tissues. PMID- 16270626 TI - Assessment of venous thromboembolism risk and the benefits of thromboprophylaxis in medical patients. AB - Hospitalized patients with acute medical conditions are at significant risk of venous thromboembolism (VTE): approximately 10-30% of general medical patients may develop deep-vein thrombosis or pulmonary embolism, and the latter is a leading contributor to deaths in hospital. Despite consensus-group recommendations that at-risk medical patients should receive thromboprophylaxis, there is currently no consensus as to which patients are at risk, and many patients may not receive appropriate thromboprophylaxis. This paper reviews evidence for the risk of VTE associated with different medical conditions and risk factors, and presents a risk-assessment model for risk stratification in medical patients. Medical conditions associated with a moderate to high risk of VTE include cardiac disease, cancer, respiratory disease, inflammatory bowel disease, and infectious diseases. Importantly, analyses of data from the MEDENOX study show that thromboprophylaxis significantly reduces the risk of VTE in these patient subgroups. Risk factors in medical patients include a history of VTE, history of malignancy, increasing age, thrombophilia, prolonged immobility, and obesity. These medical conditions and risk factors are included in a risk assessment model which is hoped will provide a simple means of assisting clinicians in deciding whether thromboprophylaxis should be used in an individual patient. PMID- 16270627 TI - Anti-factor VIII antibodies: a 2005 update. AB - The development of anti-factor VIII (FVIII) antibodies is currently one of the most serious complications in the treatment of haemophilia A patients. Numerous studies in literature report on their epitope specificity, their mechanism of FVIII inactivation, and their relationship with FVIII genetic alterations. During the last two years, however, a particular effort has been made to better understand their generation, with particular emphasis on the interplay of T cells and B cells specific for FVIII and the generation of anti-FVIII antibodies. Moreover, novel strategies to improve the management or treatment of patients with anti-FVIII antibodies have been recently proposed: the use of less immunogenic engineered recombinant FVIII molecules, neutralization of inhibitors by blocking their deleterious activity either by low molecular weight peptide decoys or by anti-idiotypic antibodies, and attempts to suppress the T-cell response involved in the antibody formation. All of these represent promising therapeutic approaches. This review attempts to sum up current knowledge of the nature and properties of anti-FVIII antibodies, their mechanism of action, their neutralization by anti-idiotypic antibodies, and the role of T cells in FVIII inhibitor formation. In the final part, some of the new strategies susceptible to improve the management or the eradication of anti-FVIII antibodies are presented. PMID- 16270628 TI - Nitric oxide differentially regulates proliferation and mobilization of endothelial progenitor cells but not of hematopoietic stem cells. AB - To investigate the role of nitric oxide in controlling endothelial progenitor (EPC) and hematopoietic stem cell (HSC) mobilization, wild-type mice, L-NAME treated WT and eNOS-/- mice received either PBS or G-CSF for 5 days. Under unstimulated conditions bone marrow of either L-NAME treated WT and eNOS-/- mice, representing acute and chronic NO-deficiency, showed higher CD34(+)Flk-I+ EPC numbers compared to their WT littermates. Furthermore, CD34(+)Flk-I+ progenitors under NO-deficient conditions showed a higher cell turn over since the proliferation and apoptosis activity under in vivo as well as in vitro conditions were enhanced. In line with this finding bone marrow derived EPC differentiation towards endothelial cells was modulated in an NO-dependent manner. Administration of G-CSF resulted in an increase of EPC within the bone marrow of WT animals with a consecutive release of these cells into the peripheral circulation. Under NO deficient conditions G-CSF failed to increase EPC numbers. In contrast, the HSC population c-kit(+)Lin- was not influenced by nitric oxide. Thus, NO differentially supports the mobilization of the endothelial committed progenitor subpopulation in bone marrow but does not have an effect on HSC in vivo. PMID- 16270629 TI - VKORC1 haplotypes and their impact on the inter-individual and inter-ethnical variability of oral anticoagulation. AB - In order to elucidate the role of VCORC1 sequence variants in warfarin sensitivity, we established a complete SNP map of the VKORC1 gene locus in 200 blood donors from Western Germany. Nearly all of the genetic variability of the VKORC1 gene in Europeans is reflected by three main haplotypes. Recently described polymorphisms associated with low warfarin dose requirement (dbSNP:rs9934438; dbSNP:rs17878363) were found in complete linkage disequilibrium with the VKORC1*2 haplotype. In two patient cohorts of European origin with either increased coumarin sensitivity (n= 14) or partial coumarin resistance (n=36) the VKORC1*2 frequency varied highly significant between the two groups and also when compared to 200 blood donor controls (coumarin sensitive 96%, coumarin resistant 7%, controls 42%) thus demonstrating a strong association between these two phenotypes and the VKORC1 haplotype (p = 1.6 x 10(-8) for coumarin sensitive and p = 1.9 x 10(-8) for coumarin resistant). Analysis of database derived VKORC1 genotypes of African Americans and Chinese revealed that haplotype frequencies in these populations differ significantly from the European sample (for VKORC1*2: Europeans 42%, Chinese 95%, African Americans 14%). These observations suggest VKORC1 as principal genetic modulator of the ethnic differences in warfarin response. Since hereditary pharmacodynamic (VKORC1) and pharmacokinetic (CYP2C9) factors account for up to 50% of the inter-individual variability of the warfarin response, these genetic markers may serve as clinically relevant predictors of warfarin dosing in future studies. PMID- 16270631 TI - Tissue factor pathway inhibitor and the risk of recurrent venous thromboembolism. AB - Tissue factor pathway inhibitor (TFPI) regulates factor X activation. Low TFPI is a risk factor for a first venous thrombosis. We evaluated whether low TFPI confers an increased risk of recurrent venous thromboembolism (VTE). TFPI-free antigen was measured in 611 patients with a first spontaneous VTE, and who were prospectively followed after withdrawal of anticoagulation. The endpoint was symptomatic recurrent VTE. The relative risk (RR) of recurrence increased from 1.0 (95% CI 0.4-2.6) in patients with TFPI levels < or = 5th percentile to 2.7 (95% CI 1.0-7.4) in patients with levels < or = 2nd percentile as compared with higher levels. At five years, the probability of recurrence was 48.6% (95th CI 19.0-78.1) among patients with TFPI < or = 2nd percentile and 16.8% (95th CI 13.8 19.8) among those with higher levels (p=0.04). Compared to patients with wild type factor V and high TFPI, the RR of recurrence was 1.1 (95% CI 0.7-1.7) in patients with factor V Leiden and high TFPI, 2.3 (95% CI 0.6-9.5) in patients with wild type factor V and low TFPI and 3.5 (95% CI 0.9-14.3) in patients with factor V Leiden and low TFPI. In a multivariate analysis,the high risk of recurrence in carriers of factor V Leiden and low TFPI slightly decreased [RR 2.8 (95% CI 0.6-9.5)]. We conclude that thrombosis patients with low levels of free TFPI are at an increased risk of recurrent VTE. PMID- 16270630 TI - Site-directed mutagenesis of coumarin-type anticoagulant-sensitive VKORC1: evidence that highly conserved amino acids define structural requirements for enzymatic activity and inhibition by warfarin. AB - Coumarin and homologous compounds are the most widely used anticoagulant drugs worldwide. They function as antagonists of vitamin K, an essential cofactor for the posttranslational gamma-glutamyl carboxylation of the so-called vitamin K dependent proteins. As vitamin K hydroquinone is converted to vitamin K epoxide (VKO) in every carboxylation step, the epoxide has to be recycled to the reduced form by the vitamin K epoxide reductase complex (VKOR). Recently, a single coumarin-sensitive protein of the putative VKOR enzyme complex was identified in humans (vitamin K epoxide reductase complex subunit 1, VKORC1). Mutations in VKORC1 result in two different phenotypes: warfarin resistance (WR) and multiple coagulation factor deficiency type 2 (VKCFD2). Here,we report on the expression of site-directed VKORC1 mutants, addressing possible structural and functional roles of all seven cysteine residues (Cys16, Cys43, Cys51, Cys85, Cys96, Cys132, Cys135), the highly conserved residue Ser/Thr57, and Arg98, known to cause VKCFD2 in humans. Our results support the hypothesis that the C132-X-X-C135 motif in VKORC1 comprises part of the redox active site that catalyzes VKO reduction and also suggest a crucial role for the hydrophobic Thr-Tyr-Ala motif in coumarin binding. Furthermore, our results support the concept that different structural components of VKORC1 define the binding sites for vitamin K epoxide and coumarin. PMID- 16270632 TI - Tissue plasminogen activator extravasated through the cerebral vessels: evaluation using a rat thromboembolic stroke model. AB - Neurotoxic effects of endogenous tissue plasminogen activator (tPA) have recently been reported. Employing a rat model of thromboembolic stroke, we evaluated the extent and degree of extravasation of exogenous tPA administered for the purpose of fibrinolysis. In a thromboembolic model using Sprague-Dawley rats, focal cerebral ischemia was induced at the territory of the middle cerebral artery (MCA). Early reperfusion was induced by administering tPA (10 mg/kg) intravenously at 30 minutes after the onset of ischemia. Extravasated tPA was evaluated by immunohistochemistry, and the concentration of tPA in the brain tissue was quantified by enzyme-linked immunosorbent assay methods. The integrity of the blood-brain barrier (BBB) was examined electronmicroscopically. In a thread model of transient ischemia, reperfusion was induced without tPA administration at 30 minutes or 2 hours after the onset of ischemia, and the tPA content of the brain was quantified. In the rats with thromboembolic stroke, extravasation of tPA was observed at the territory of the MCA. Both the endogenous and exogenous tPA contents were 3.5 +/- 1.6 ng/ml of homogenized brain in saline. Electronmicroscopically, mild ischemic changes were observed, although the integrity of the BBB was preserved. In the thread model rats, the endogenous tPA contents of the ischemic hemisphere were 0.9 +/- 0.1 and 1.0 +/- 0.2 ng/ml in the 30-minute and 2-hour ischemia groups, respectively, and were significantly lower than the tPA contents in the thromboembolic stroke rats (p<0.01). The present findings indicate that significant extravasation of exogenous tPA occurs through the cerebral vessels even though early reperfusion is induced. PMID- 16270633 TI - High dose dexamethasone increases circulating P-selectin and von Willebrand factor levels in healthy men. AB - Although glucocorticoids are widely used in a number of inflammatory disorders associated with endothelial and platelet activation, their effect on the endothelium and platelets in humans remain poorly defined. Hence,we measured changes of a specific endothelial cell marker (von Willebrand factor [vWF]) and of a platelet marker (soluble P-selectin) by infusing therapeutic doses of dexamethasone (0.04 mg/kg and 1.0 mg/kg b.i.d on two days) or placebo into nine healthy men. Venous citrated plasma was obtained before infusion, and at 24 and 48 h. Compared to baseline levels, we found increased levels of vWF at both time points at the higher dose (p=0.011). Plasma levels of sP-selectin rose at 48 h after the high dose (p=0.017). Human umbilical endothelial cells were cultured in the presence or absence of dexamethasone (0, 0.01, 1 microM), to determine the possible mechanism for the increase in vWF. The vWF-mRNA levels as quantified by RT-PCR increased 2-fold (p<0.05), and the vWF-concentrations in cell lysates increased by 38% (p<0.05), whereas the vWF-concentrations in the supernatants were unaffected. In summary, high dose DEXA increases sP-selectin and vWF. The probable underlying mechanism for the latter was a DEXA induced up-regulation of vWF-mRNA transcription. Together, this indicates that high dose glucocorticoids may enhance haemostasis, which could be beneficial under certain conditions, but which may also contribute to adverse vascular events by increasing platelet activation and vWF dependent thrombosis. PMID- 16270634 TI - Safety and pharmacokinetics of recombinant factor XIII in healthy volunteers: a randomized, placebo-controlled, double-blind, multi-dose study. AB - Factor XIII (FXIII) is a plasma transglutaminase that covalently cross-links fibrin proteins to one another and to other proteins, increasing the mechanical strength of blood clots. Endogenous FXIII is the final enzyme in the clotting cascade and circulates as a heterotetramer comprising 2 FXIII-A subunits and 2 FXIII-B subunits. Recombinant human FXIII A2 (rFXIII) homodimer is produced in Saccharomyces cerevisiae. Upon injection, rFXIII combines with the free FXIIIB subunit that circulates in excess to form the rA2B2 tetramer. In this placebo controlled, double-blind, multi-dose study, the safety, pharmacokinetics, and pharmacodynamics of rFXIII were studied in 24 healthy volunteers, who were randomized in 2 cohorts of 12 subjects each. In each cohort, 8 subjects received 5 daily intravenous doses of rFXIII (10 or 25 U/kg), and 4 subjects received placebo. Recombinant FXIII was well tolerated. No deaths or serious adverse events occurred. The type and frequency of adverse events showed no pattern or dose response. No clinically significant changes in haematology, serum chemistry, or urinalysis laboratory values were observed. No clinical coagulopathy or thrombosis was observed. Recombinant FXIII did not produce any anti-yeast or anti FXIII antibodies. After 5 daily doses of rFXIII, accumulation indices indicated a 3 to 4fold accumulation of rFXIII in plasma. The elimination half-life, estimated for rFXIII as the heterotetramer, ranged from 228-346 hours for the 10U/kg dose group and 167-197 hours for the 25U/kg dose group. The safety, pharmacokinetic, and immunogenic profile of rFXIII suggests it may have potential use in patients with congenital or acquired FXIII deficiency. PMID- 16270635 TI - Effects of antithrombin and heparin cofactor II levels on anticoagulation with Intimatan. AB - Heparin is the current mainstay drug for anticoagulation during cardiac surgery, but it requires normal levels of antithrombin (AT) for optimal anticoagulation. Heparin anticoagulation may be less effective in cardiac surgical patients because of decreased AT levels due to the prolonged heparin therapy. Therefore, other anticoagulants that would work well in AT deficient patients may be more desirable. One such agent currently being evaluated is Intimatan, which catalyzes heparin cofactor II (HCII) dependent inhibition of thrombin. In the current in vitro study we examined the effects of Intimatan (20 microg/ml), heparin (0.25 U/ml), or both drugs in combination on thrombin generation in plasma with decreasing levels of AT, HCII or both cofactors, using a novel method based on the continuous measurement of thrombin generation. For the study, we collected blood samples from healthy volunteers, isolated platelet poor plasma by centrifugation and mixed it with AT, HCII, or AT-HCII deficient plasma samples to achieve different levels of AT, HCII and AT-HCII. Thrombin generation was inhibited equally well with heparin or Intimatan when the level of their respective cofactors was within the normal range. With decreasing levels of AT or HCII, heparin and Intimatan became less effective in thrombin inhibition, respectively. With the absence of both cofactors, neither agent alone or in combination had any effect on thrombin generation. We conclude that Intimatan may be an effective adjunct to heparin therapy under low AT conditions. PMID- 16270636 TI - Prevention of fatal pulmonary embolism and mortality in surgical patients: a randomized double-blind comparison of LMWH with unfractionated heparin. AB - The incidences of fatal pulmonary embolism and death in surgical patients receiving low-molecular-weight heparin thromboprophylaxis have not been previously determined in large, adequately designed clinical trials and information on the relative efficacy and safety of unfractionated and low molecular-weight heparin in preventing these clinical endpoints is not available. In a double-blind study, 23078 surgical patients randomly received the low molecular-weight heparin, certoparin (3000 anti Xa IU) subcutaneously once-daily, or unfractionated heparin (5000 IU) subcutaneously three-times daily, for a minimum of 5 days. The primary outcome measure, autopsy-proven fatal pulmonary embolism recorded up to 14 days after the end of prophylaxis, occurred in 0.152% (95% confidence interval (CI) 0.10, 0.20%; 35 of 23078 patients) of cases, with no significant difference between the certoparin-treated patients (0.147% (95% CI 0.077, 0.217%; 17 of 11542 patients) and patients treated with unfractionated heparin (0.156% (95% CI 0.084, 0.228%; 18 of 11,536 patients, P=0.868). The autopsy rate was 70.2%. Comparing mortality, there was no significant difference between the groups (1 .44% [166 of 11542 certoparin patients] versus 1.27% [146 of 11536 unfractionated heparin patients]; P=0.279). The safety profiles of both treatment groups were similar. Once-daily certoparin and three-times daily unfractionated heparin are equally effective and safe in reducing fatal pulmonary embolism and death to low levels in surgical patients and mirror the findings of comparative efficacy studies using surrogate endpoints. PMID- 16270637 TI - Ximelagatran for the secondary prevention of venous thromboembolism: a complementary follow-up analysis of the THRIVE III study. AB - In the randomized, double-blind THRIVE III trial, the oral direct thrombin inhibitor ximelagatran (24 mg twice daily) significantly reduced the incidence of recurrent venous thromboembolism (VTE) versus placebo over 18 months or until premature study drug discontinuation. A complementary follow-up analysis (intention-to-treat) was conducted post-study to evaluate the cumulative risks of locally-confirmed recurrent VTE and death (Kaplan-Meier procedure) over the full 18-month study period, regardless of whether patients discontinued study drug prematurely. Hazard ratios (HRs) between treatments were estimated using Cox proportional hazard modeling. Of 612 and 611 patients receiving ximelagatran and placebo, respectively, 149 and 181 discontinued treatment prematurely. Of these discontinuations, further information could not be collected for 14 and 13 patients in the ximelagatran and placebo groups, respectively; among the remaining patients, four VTE events and four deaths occurred in the ximelagatran group, and one VTE event and five deaths occurred in the placebo group. The resulting cumulative risks of VTE (3.2% vs. 12.7%; HR 0.21; 95% confidence interval [CI], 0.12, 0.36; P < 0.0001) and pulmonary embolism (0.8% vs. 5.2%; HR 0.13; 95% CI 0.04, 0.36; P < 0.0001) were significantly lower in the ximelagatran than in the placebo group over 18 months. Death from any cause over 18 months occurred in 10 and 12 patients, respectively (HR 0.83;95% CI 0.36, 1.93; P = 0.7). This complementary follow-up analysis confirms the benefit of oral ximelagatran 24 mg twice daily, administered without coagulation monitoring or dose adjustment, for the long-term secondary prevention of VTE. PMID- 16270638 TI - The post-thrombotic syndrome: incidence and prognostic value of non-invasive venous examinations in a six-year follow-up study. AB - The ability to predict severity of the post-thrombotic syndrome (PTS) early after acute deep-vein thrombosis (DVT) is limited. The aim of our study was to examine the incidence of PTS prospectively and to evaluate the predictive value of non invasive venous examinations shortly after DVT for the development of PTS. In 93 patients with DVT thrombosis score (TS), reflux, venous outflow resistance (VOR) and calf muscle pump dysfunction (CMP) were examined prospectively. After one, two and six years patients were evaluated for PTS using the clinical scale of the CEAP-classification (PTS present > or = 3 on a scale from 0 to 6). Area under the curves (AUC) were used to evaluate the predictive value of the non-invasive examinations at one and three months after diagnosis of DVT for future PTS. The cumulative incidence of PTS increased from 49% (32/65) after one year to 55% (36/65) and 56% (27/48) after two and six years, whereas the incidence of patients with PTS class 4 progressed from 20% after two years to 33% after six years. The prognostic value to predict PTS was highest for the combination of TS, VOR and reflux measured three months after diagnosis and showed an AUC of 0.77 (0.65-0.90) for PTS after one year. In conclusion, the incidence of PTS after DVT did not increase significantly after one year, whereas during longer follow-up the severity of PTS rose in patients with PTS. Moreover, measurement of TS, VOR and reflux three months after DVT could predict, with reasonable accuracy, the risk of PTS after one year of follow-up. PMID- 16270639 TI - Agonists of toll-like receptor (TLR)2 and TLR4 are unable to modulate platelet activation by adenosine diphosphate and platelet activating factor. AB - Inappropriate platelet activation is a feature of acute and chronic diseases such as disseminated intravascular coagulation (DIC) and atherosclerosis. Since proinflammatory microbial-derived agonists can be involved in the pathogenesis of these diseases, we examined the potential role of TLR4 (mediating responses to LPS) and TLR2 (which responds to bacterial lipopeptides) in platelet activation. Our data suggested low-level expression of TLR2 and TLR4 on platelets, determined by flow cytometry, and we also observed expression of TLR4 on a megakaryocytic cell line by both flow cytometry and immunohistochemistry. Stimulation of the platelets with the TLR4 agonist LPS, and the synthetic TLR2 agonist Pam3CSK4, resulted in no platelet aggregation, no increase in CD62P surface expression and no increase in the cytosolic concentration of Ca2+. The TLR agonists were also unable to directly activate platelets primed with epinephrine, or pretreated with a low concentration of ADP or PAF. Pretreatment of platelets with LPS or Pam3CSK4 also failed to modulate the platelet response to submaximal concentrations of the classical platelet agonists ADP and PAF. We conclude that the TLR agonists LPS and Pam3CSK4 have no direct effect on platelet activation and that platelet TLRs may be a remnant from megakaryocytes. TLR2 and TLR4 agonists are thought to have a significant role in diseases such as atherosclerosis and DIC, but our research suggests that this is through a mechanism other than direct platelet activation or by modification of platelet responses to other agonists. PMID- 16270640 TI - Transient receptor potential protein subunit assembly and membrane distribution in human platelets. AB - We have previously suggested that the human homologue of the Drosophila transient receptor potential protein, TRPC1, is involved in conducting store-operated Ca2+ entry (SOCE) in human platelets since an antibody raised against the pore-forming region of TRPC1 inhibited SOCE. Here we have investigated plasma membrane expression of TRPC1 in human platelets and have probed for the presence of other TRPC proteins in these cells. Biotinylation revealed the presence of TRPC1 in the plasma membrane of resting platelets. Surface expression was not detectibly changed following Ca2+ store depletion or stimulation with thrombin. Western blotting demonstrated the presence of TRPC1, TRPC3, TRPC4, TRPC5 and TRPC6 in platelet lysates. TRPC1, TRPC4 and TRPC5 coimmunoprecipitated, as did TRPC3 and TRPC6. TRPC1, TRPC4 and TRPC5 were associated with detergent-resistant platelet membranes, from which they were partially released when the cells were cholesterol-depleted using methyl-beta-cyclodextrin. The distributions of TRPC3 and TRPC6 between soluble and membrane fractions were not affected by methyl-beta cyclodextrin treatment. These results suggest that TRPC1, TRPC4 and TRPC5 form a heteromultimer associated with platelet lipid raft domains, whereas TRPC3 and TRPC6 associate independently of lipid rafts. PMID- 16270641 TI - Exercise paradoxically modulates oxidized low density lipoprotein-induced adhesion molecules expression and trans-endothelial migration of monocyte in men. AB - Physical exercise can affect the risk of cardiovascular disease. Oxidized-low density lipoprotein (ox-LDL) promotes transendothelial migration (TEM) of monocyte, thereby accelerating the pathogenesis of atherosclerosis. This study investigated how exercise intensity affects monocyte/EC interactions under ox-LDL mediated condition. Light- (LIE), moderate- (MIE) and high- (HIE) intensity exercise (i.e., 40%, 60%, and 80% VO2max, respectively) on a bicycle ergometer in 18 sedentary healthy men were performed on three separate occasions. Before and immediately after exercise, ox-LDL-promoted expressions of monocyte adhesion molecules and TEM of monocyte, as well as oxidation of LDL and amounts of soluble adhesion molecules in plasma were measured. Analytical results showed that (1) ox LDL furthered monocyte L-selectin shedding and Mac-1 expression, and an attendant increase in TEM of monocyte, while treating the monocyte with Mac-1 antibody inhibited the ox-LDL-promoted TEM of monocyte; (2) under ox-LDL-treated condition, MIE increased monocyte Mac-1 and LFA-1 expressions, enhancing the TEM of monocyte, whereas HIE downregulated monocyte Mac-1 expression, suppressing the TEM of monocyte; (3) LIE decreased basal LFA-1 expression as well as basal and ox LDL-promoted TEM of monocyte; and (4) MIE and HIE, but not LIE, elevated plasma ox-LDL level, while there were no significant changes in sL-selectin, sE selectin, sICAM-1, and sVCAM-1 following these exercises. Therefore, we conclude that monocyte activation and subsequent TEM promoted by ox-LDL are changed by short-term exercise in an intensity-dependent manner. These findings provide a new insight into the may aid the development of suitable exercise intensity enable people to prevent early atherogenesis. PMID- 16270642 TI - Microparticles from apoptotic vascular smooth muscle cells induce endothelial dysfunction, a phenomenon prevented by beta3-integrin antagonists. AB - Fragile atherosclerotic plaques are rich in apoptotic smooth muscle cells (SMCs) and macrophages, generating microparticules (MPs) which accumulate locally and may be released in blood in case of mechanical or spontaneous plaque disruption. Besides being highly procoagulant, this material may interact with downstream endothelium. Using a model of mouse aorta vaso-reactivity, we have investigated the effects of apoptotic MPs prepared in vitro from Fas-ligand sensitive SMCs. Short-term preincubation of aorta rings with the MPs dose-dependently reduced the vasodilatory response to acetylcholine dependent on the endothelium. This effect was prevented by the addition of abxicimab or eptifibatide, indicating a role for a beta3 integrin in this process. We further investigated its mechanism using cultured endothelial cells. The MPs were found to bind to the cells and to inhibit the production and the release of nitric oxide (NO) in response to bradykinin. This phenomenom was redox sensitive, independent of the generation of activated coagulation proteases, and was abrogated when the MPs were pretreated by trypsin. The metabolic effects of MPs were prevented by the addition of eptifibatide. Taken together, these results suggest a potential, platelet independent, mechanism for the improvement of microvascular perfusion observed with beta3-integrin antagonists. PMID- 16270643 TI - Indication of a role of plasminogen activator inhibitor type I in protecting murine fibrosarcoma cells against apoptosis. AB - In a number of cancer types high tumor tissue levels of plasminogen activator inhibitor type 1 (PAI-1) protein are strongly associated with shorter cancer patient survival. This association has been intriguing since PAI-1 is known to inhibit urokinase plasminogen activator (uPA) that converts plasminogen to plasmin, which is actively involved in tumor progression and invasion. In order to further explore the biological role of PAI-1 in cancer, we have prepared fibroblasts from PAI-1 gene deficient mice and from their wild type littermates. From these fibroblasts fibrosarcoma cell lines were established and characterized. Both types of fibroblasts underwent spontaneous transformation as indicated by aneuploidy, immortalization, clonogenicity in soft agar and tumor formation in vivo. While both PAI-1 deficient and PAI-1 expressing cell lines showed similar proliferation rates in vitro, cells devoid of PAI-1 were significantly more sensitive to apoptotic stimuli. When inoculated subcutaneously into nude mice PAI-1 expressing cells rapidly established tumors, while PAI-1 deficient cells had a significantly longer lag-phase before they started to grow (p<0.0001). The present study suggests that PAI-1, besides its uPA inhibiting function, has a role in cancer progression by protecting tumor cells from undergoing apoptosis. PMID- 16270645 TI - Detection of known haemophilia B mutations and carrier testing by microarray. AB - The molecular basis of haemophilia B is heterogeneous and many mutations of the Factor IX (FIX) gene have been characterised. Using the allele-specific arrayed primer extension (AS-APEX) technology, we have designed a FIX array to simultaneously analyse 69 mutations found in British, Thai and Chinese patients. This technology overcomes the problem of multiple reverse dot-blot analysis and has a 100% accuracy in the detection of both affected subjects and carriers in families with known mutations. In seven unknown mutations from Thailand, the array could detect the specific mutation in five and in the remainders the normal primer at specific spots failed to extend due to a mutation a few nucleotides upstream, thus allowing their identification. Hence this FIX array can detect 53% of the 2891 mutation entries in the FIX database. Each of the microarray slide can be used for three different test samples and would be useful for carrier testing for common mutations and prenatal diagnosis. It is simpler and more cost effective than genome sequencing and would be particularly useful in laboratories with limited technical capabilities. PMID- 16270644 TI - Evaluation of perioperative fatal pulmonary embolism and death in cancer surgical patients: the MC-4 cancer substudy. AB - Cancer patients undergoing surgery are at a high risk of venous thromboembolism, but few studies have described the rate of autopsy-confirmed fatal pulmonary embolism after heparin thromboprophylaxis. In a post hoc analysis of a randomized study (MC-4), which compared the efficacy and safety of certoparin (3000 anti-Xa IU, subcutaneously, once-daily) with unfractionated heparin (5000 IU, subcutaneously, three-times daily) in 23078 patients undergoing surgery lasting more than 30 min, the incidence of autopsy-confirmed fatal pulmonary embolism, death and bleeding in the cancer patients (n=6124) was compared with non-cancer patients (n=16954). Fatal pulmonary embolism was significantly more frequent in cancer patients (0.33% [20/6124]) than in non-cancer patients (0.09% [15/16954], relative risk (RR), 3.7 [95% confidence intervals (CI), 1.80, 7.77], p=0.0001) at 14 days post-prophylaxis. Perioperative mortality was also significantly higher in cancer patients than in noncancer patients (3.14% [192/6124] vs. 0.71% [120/16954], RR, 4.54 [95% CI, 3.59, 5.76], p=0.0001), as were blood loss (p<0.0001), and transfusion requirements (p<0.0001). Prevention of venous thromboembolism in cancer surgical patients remains a clinical challenge. PMID- 16270646 TI - Acquired Glanzmann's thrombasthenia variant and immune thrombocytopenia in a renal transplant recipient receiving tacrolimus. PMID- 16270647 TI - Superior sagittal sinus thrombosis after lumbar puncture in a patient with T-cell lymphoblastic lymphoma: role of the prothrombin G20210A mutation and 4G/4G genotype. PMID- 16270648 TI - Molecular characterization of factor IX gene mutations in 53 patients with haemophilia B in India. PMID- 16270649 TI - High stability of blood samples for flow cytometric analysis of VASP phosphorylation to measure the clopidogrel responsiveness in patients with coronary artery disease. PMID- 16270650 TI - The polyphenols quercetin and catechin synergize in inhibiting platelet CD40L expression. PMID- 16270651 TI - Effect of compliance and dosage adaptation of long term aspirin on platelet function with PFA-100 in patients after myocardial infarction. PMID- 16270652 TI - Pre-procedural levels of erythrocyte sedimentation rate (ESR) and risk of clinical restenosis in patients with percutaneous coronary intervention and coronary stent placement. PMID- 16270653 TI - Hypersensitivity to the pentasaccharide fondaparinux in patients with delayed type heparin allergy. PMID- 16270654 TI - Metabolic engineering. AB - Metabolic engineering is a powerful methodology aimed at intelligently designing new biological pathways, systems, and ultimately phenotypes through the use of recombinant DNA technology. Built largely on the theoretical and computational analysis of chemical systems, the field has evolved to incorporate a growing number of genome scale experimental tools. This combination of rigorous analysis and quantitative molecular biology methods has endowed metabolic engineering with an effective synergism that crosses traditional disciplinary bounds. As such, there are a growing number of applications for the effective employment of metabolic engineering, ranging from the initial industrial fermentation applications to more recent medical diagnosis applications. In this review we highlight many of the contributions metabolic engineering has provided through its history, as well as give an overview of new tools and applications that promise to have a large impact on the field's future. PMID- 16270656 TI - Plant cells: secondary metabolite heterogeneity and its manipulation. AB - This chapter proposes the concept of rational manipulation of secondary metabolite heterogeneity in plant cell cultures. The heterogeneity of plant secondary metabolites is a very interesting and important issue because these structure-similar natural products have different biological activities. Both taxoids and ginsenosides are two kinds of preeminent examples in the enormous reservoir of pharmacologically valuable heterogeneous molecules in the plant kingdom. They are derived from the five-carbon precursor isopentenyl diphosphate, produced via the mevalonate or the non-mevalonate pathway. The diterpenoid backbone of taxoids is synthesized by taxadiene synthase and the triterpenoid backbone of ginsenosides is synthesized by dammarenediol synthase or beta-amyrin synthase. After various chemical decorations (oxidation, substitution, acylation, glycosylation, benzoylation, and so on) mediated by P450-dependent monooxygenases, glycosyltransferases, acyltransferases, benzoyltransferases, and other enzymes, the terpenoid backbones are converted into heterogeneous taxoids and ginsenosides with different bioactivities. Although detailed information about accumulation and regulation of individual taxoids or ginsenosides in plant cells is still lacking, remarkable progress has recently been made in the structure and bioactivity identification, biosynthetic pathway, manipulation of their heterogeneity by various methodologies including environmental factors, biotransformation, and metabolic engineering in cell/tissue cultures or in plants. Perspectives on a more rational and efficient process to manipulate production of desired plant secondary metabolites by means of metabolic engineering and "omics"-based approaches (e.g., functional genomics) are also discussed. PMID- 16270655 TI - Microbial isoprenoid production: an example of green chemistry through metabolic engineering. AB - Saving energy, cost efficiency, producing less waste, improving the biodegradability of products, potential for producing novel and complex molecules with improved properties, and reducing the dependency on fossil fuels as raw materials are the main advantages of using biotechnological processes to produce chemicals. Such processes are often referred to as green chemistry or white biotechnology. Metabolic engineering, which permits the rational design of cell factories using directed genetic modifications, is an indispensable strategy for expanding green chemistry. In this chapter, the benefits of using metabolic engineering approaches for the development of green chemistry are illustrated by the recent advances in microbial production of isoprenoids, a diverse and important group of natural compounds with numerous existing and potential commercial applications. Accumulated knowledge on the metabolic pathways leading to the synthesis of the principal precursors of isoprenoids is reviewed, and recent investigations into isoprenoid production using engineered cell factories are described. PMID- 16270657 TI - Model-based inference of gene expression dynamics from sequence information. AB - A dynamic model of prokaryotic gene expression is developed that makes considerable use of gene sequence information. The main contribution arises from the fact that the combined gene expression model allows us to access the impact of altering a nucleotide sequence on the dynamics of gene expression rates mechanistically. The high level of detail of the mathematical model is considered as an important step towards bringing together the tremendous amount of biological in-depth knowledge that has been accumulated at the molecular level, using a systems level analysis (in the sense of a bottom-up, inductive approach). This enables to the model to provide highly detailed insights into the various steps of the protein expression process and it allows us to access possible targets for model-based design. Taken as a whole, the mathematical gene expression model presented in this study provides a comprehensive framework for a thorough analysis of sequence-related effects on the stages of mRNA synthesis, mRNA degradation and ribosomal translation, as well as their nonlinear interconnectedness. Therefore, it may be useful in the rational design of recombinant bacterial protein synthesis systems, the modulation of enzyme activities in pathway design, in vitro protein biosynthesis, and RNA-based vaccination. PMID- 16270658 TI - Trends and challenges in enzyme technology. AB - Several major developments took place in the field of biocatalysis over the past few years. These include the invention of directed evolution as an extremely useful method for biocatalyst improvement on the molecular level in combination with high-throughput screening systems, methods for accessing "nonculturable" biodiversity using metagenome approaches and progress in sequence-based biocatalyst discovery. In addition, new carriers and tools for immobilization of enzymes have been developed. For the synthesis of optically active compounds impressive examples using new enzymes and major progress in dynamic kinetic resolutions of racemates took place. These achievements are summarized in this review. PMID- 16270659 TI - Regioselective glycosylation: synthesis of alpha-indoline nucleosides. AB - Novel indoline ribonucleosides with the alpha-N-glycoside configuration are synthesized with very high regioselectivity in 90-96%yield, using TMS protected indolines and 2,3-O-(1-methylethylidene)-5-O-(triphenylmethyl)-alpha/beta-D ribofuranose. The structures of these ribonucleosides were elucidated with X-ray crystallography as well as 2D (NOESY, COSY, and HMQC) NMR spectroscopy. PMID- 16270660 TI - Zwitterionic oligonucleotides: a study on binding properties of 2'-O-aminohexyl modifications. AB - 2'-O-Aminohexyl side chains provide excellent conditions for zwitterionic interstrand and intrastrand interactions of oligonucleotides. 2'-O-Aminoalkylated phosphoramidites of adenosine and uridine were synthesized and incorporated in increasing number into homo adenosine and homo uridine/thymidine dodecamers, respectively. CD spectra of these dodecamers with complementary sense DNA exhibited a B-DNA type structure. While duplex stability values of all tested oligonucleotides were lower than those of the native oligonucleotides, they were significantly higher than those of 2'-O-heptyl modified oligonucleotides. The destabilization amounted to 0.9, 1.5, and 2.7 degrees C per modification for 2'-O aminohexyl adenosine, 2'-O-aminohexyl uridine, and 2'-O-heptyl adenosine substitutions. These findings are pointing to a duplex stabilizing effect of the interaction of side chain amino groups with backbone phosphoric acid. PMID- 16270661 TI - Synthesis of phosphonate analogues of the antiviral cyclopropane nucleoside A 5021. AB - A series of phosphonate analogues of the antiviral cyclopropane nucleoside A-5021 were synthesized from (1S*, 7R*)-3,5-dioxa-4,4-diphenylbicyclo[5. 1.0]octane-l methanol by a 10-step process. In contrast to the potent antiherpetic activity of A-5021, they were all devoid of antiviral activity. PMID- 16270662 TI - Crystal structure and conformation of 8-(2-hydroxyethylamino) and 8-(pyrrolidin-1 yl) adenosines. AB - In the course of investigation of 8-alkylamino substituted adenosines, the title compounds were synthesized as potential partial agonists for adenosine receptors. The structure determination of these compounds was carried out with the X-ray crystallography study. Crystals of 8-(2-hydroxyethylamino)adenosine are monoclinic, space group P 2(1); a = 7.0422(2), b = 11.2635(3), c = 8.9215(2) A, beta = 92.261(1) degrees, V = 707.10(3) A3, Z = 2; R-factor is 0.0339. The nucleoside is characterized by the anti conformation; the ribose ring has the C(2')-endo conformation and gauche-gauche form across C(4')-C(5') bond. The molecular structure is stabilized by intramolecular hydrogen bond of N-HO type. Crystals of 8-(pyrrolidin-1-yl)adenosine are monoclinic, space group C 2; a = 19.271(1), b = 7.3572(4), c = 11.0465(7) A, beta = 103.254(2), V = 1524.4(2) degrees A3, Z = 4; R-factor is 0.0498. In this compound, there is syn conformation of the nucleoside; the ribose has the C(2')-endo conformation and gauche -gauche form across C(4')- C(5') bond. The molecular structure is stabilized by intramolecular hydrogen bond of O-HN type. For both compounds, the branching net of intermolecular hydrogen bonds occur in the crystal structures. PMID- 16270664 TI - Formation of Au(III)-DNA coordinate complex by laser ablation of Au nanoparticles in solution. AB - We discovered that an Au(III)-DNA coordinate complex, Au(III)(DNA-base)2(amine)L, are formed by laser ablation of Au nanoparticles in an aqueous solution containing DNA molecules in the presence of amines and multi-valent cations, where L represents an unknown ligand (either amine or water). Optical absorption spectrum of the solution after laser ablation exhibited a 360 nm absorption peak assined to ligand-->Au(III) charge transfer (LMCT) band of the coordinate complex. The complex is considered to be formed as follows: (1) the DNA molecules are neutralized by binding the multi-valent cations to their negatively charged phosphate groups, and adsorbed on the surface of the Au nanoparticles by a hydrophobic interaction, (2) Au(III) ions are liberated from the Au nanoparticles by laser ablation, and (3) an Au(III) ion reacts with amine and two DNA bases of a DNA molecule into an Au(III)(DNA-base)2(amine)L. PMID- 16270663 TI - N4-hydroxycytosine dioxolane nucleosides and their activity against hepatitis B virus. AB - Novel racemic, D- and L-beta-dioxolane N4-hydroxycytosine nucleosides have been synthesized and evaluated for their activity against hepatitis B virus. None of the synthesized nucleosides demonstrated selective anti-HBV activity. PMID- 16270665 TI - New trends in synthesis of pyrazole nucleosides as new antimetabolites. AB - Pyrazole nucleosides and condensed pyrazole nucleosides exhibit various biological activities. This article describes recent synthetic approaches to their preparation, chemical properties, biological activities, and structure activity relationships, with emphasis to selected drugs or drug candidates. Two pyrazole C-nucleoside compounds pyrazofurin (pyrazomycin) and its alpha-epimer pyrazofurin B are active components of potent antivirals approved for therapeutic use in human medicine aimed against various diseases caused by DNA viruses. PMID- 16270666 TI - New acyclonucleosides: synthesis and anti-HIV activity. AB - The synthesis of new acyclic nucleosides is described. These syntheses were accomplished by various methods: glycosylation, selective or total deprotection, oxidation/reduction, chlorination or azidation of hydroxyl groups. The compounds were characterized with NMR, mass and IR spectroscopy. Antiviral properties of these compounds were evaluated on HIV-1 infected cell lines. PMID- 16270667 TI - Synthesis of novel acyclonucleosides analogs of pyridothienopyrimidine as antiviral agents. AB - Nucleoside analogs of pyridothienopyramidines were prepared by condensing the sodium salt 2a,b with an acyclic side chain in the form of acetylated haloalkoxyalcohol, and subsequent removal of the protecting acetyl group in ammonia/methanol afforded 4a,b. The O-tosyl derivative of 4a could then be modified to azido- and amino derivatives. Reaction of the sodium salt of 2b with halo-ether, benzyl halo-ether and/or halo-thioether gave N- and S-alkylated products, 8 and 9, respectively. Coupling of 10 with the sodium salt of 2a,b gave the corresponding dioxolane derivatives 11, 13, and 14, which were treated with 80% acetic acid at room temperature to give diols 12, 15, and 16. Treatment of 16 with tosyl chloride afforded the ditosylate 17 and this could then be modified to diazido and diamino derivatives. Some of the products were screened for their biological activity. PMID- 16270668 TI - A Guide to Legal Issues in Life-Limiting Conditions. PMID- 16270669 TI - Reforming residency: modernizing resident education and training to promote quality and safety in healthcare. AB - The goal of medical residency is to provide the best clinical education for future practice, while increasing quality and safety in current and future healthcare. This goal is not being met. Traditional residency programs often continue to utilize individually oriented, shame-and-blame approaches that do not recognize the systems nature of outcomes, care, and patient safety. Appropriate substantive methods, content, and training tools are also lacking, while residents continue to labor in a poor working environment. All these factors create a system that serves no one-not the resident, the patient, or the system in which both interact. Residency reforms are proposed to address these imperative concerns. PMID- 16270670 TI - Law and public health: beyond emergency preparedness. AB - This Article examines three questions: What is public health? What is public health law? What roles can lawyers play in public health? It first describes the breadth of public health, highlighting six trends shaping its future: social determinants of health; synergy between medicine and public health; shifts in focus from external (e.g., environmental and social) to internal (behavioral) risks to health; federalization of public health law; globalization of health risks and responses; and bioterrorism. Because the domains of law that apply to public health are equally broad, the Article next offers a conceptual framework for identifying the types of laws most suitable to different public health problems. Finally, the role of lawyers in the applied field of public health law is examined, first to encourage attention to law's effect on health, even laws having little apparent relationship to health; and second, to recognize that laws intended to achieve specific health outcomes may affect broader legal principles. Lawyers have a unique role to play in ensuring that the legal principles used to promote health also preserve justice. PMID- 16270671 TI - The National Response Plan: a new framework for homeland security, public health, and bioterrorism response. AB - This Article provides a detailed overview of the new National Response Plan (NRP) with a focus on its applicability to bioterrorism and other public health emergencies. The Article highlights critical policy and legal issues left unresolved by the NRP, and offers recommendations for the resolution of those issues. The author concludes that, although the NRP is not perfect, it represents a major advance in domestic incident management and provides regular opportunities for review and revision as we learn how to best coordinate the national response to major incidents. A close working relationship between the Departments of Health and Human Services and Homeland Security should enable a unified response to bioterrorism and other public health emergencies in support of state and local efforts. PMID- 16270672 TI - The statue of security: human rights and post-9/11 epidemics. AB - Following the attacks of September 11, 2001(9/11), there have been increasingly divisive proposals for Americans to sacrifice liberty for safety. "Health and human rights" provides a much more constructive and effective framework than fear on which to base both routine public health actions and responses to public health emergencies. This can be illustrated by the failure of bioterrorism-based government initiatives at both the state and federal levels since 9/11, as well as the worldwide response to the post-9/11 SARS epidemic. Effective public health work can only occur with a population that trusts its public health officials. 9/11 can serve as a catalyst to reform public health by adopting the health and human rights framework that can move it from a fragmented state and local activity to one that is federal and coordinated, and ultimately one that is global and based on transparency, trust, science, and respect for human rights. PMID- 16270673 TI - Model conflict-of-interest policies. PMID- 16270674 TI - Sealants: basic information, advanced technology. PMID- 16270675 TI - Army dental assistants at it again. Assisting and performing community relations at 2005 National Boy Scout Jamboree. PMID- 16270676 TI - Ergonomics for the dental assistant. PMID- 16270677 TI - Evolving realities of dental practice: Care for patients with special needs. PMID- 16270678 TI - Damaged buildings and widespread environmental hazards remain in Katrina's aftermath. PMID- 16270679 TI - Craniopharyngioma. PMID- 16270680 TI - Medulloblastoma. PMID- 16270681 TI - Quality of life and behavioral follow-up study of pediatric survivors of craniopharyngioma. AB - OBJECT: The authors set out to evaluate the quality of life (QOL), social emotional functioning, and behavioral functioning of children treated surgically for craniopharyngiomas. METHODS: Twelve girls and 17 boys with a mean age at diagnosis of 8 +/- 3.8 years were surgically treated between 1985 and 1998 at the New York University Medical Center. After a mean follow-up period of 6.8 +/- 3.5 years, these 29 patients were administered either the 36-item Short Form Health Survey version 2 or the Child Health Questionnaire-Parent Form to assess QOL, as well as the Achenbach Child Behavior Checklist or Young Adult Checklist to measure social-emotional and behavioral functioning. Patients older than 19 years of age and parents of patients younger than 19 years of age reported low average overall physical QOL, with overall psychosocial QOL in the average range. Behavioral difficulties were noted, including internalizing, attention, somatic, and social difficulties. Further analyses indicated that retrochiasmatic tumor location, recurrence, and additional surgery were associated with poorer outcomes. In contrast, hydrocephalus, tumor size, and sex were not prognostic variables, and patients significantly improved as post-operative time increased. CONCLUSIONS: Attention toward late effects arising after the treatment of pediatric craniopharyngioma, including decreased postoperative physical health and behavioral functioning, is warranted. Future approaches to treatment should consider the documented effects of either gross-total resection or limited surgery followed by cranial irradiation on QOL, with specific evaluation for those with retrochiasmatic tumors, a recurrent tumor, or the need for additional surgery. Psychosocial QOL and social-emotional functioning should be maintained through ongoing counseling and education. PMID- 16270682 TI - Preoperative chemotherapy in children with high-risk medulloblastomas: a feasibility study. AB - OBJECT: The authors set out to evaluate the feasibility and effectiveness of preoperative chemotherapy in treating high-risk medulloblastomas. METHODS: Between 1997 and 2000, 21 children with high-risk medulloblastomas (M > or = 2 and/or T3b/T4 according to the Chang classification) were treated consecutively in a pilot study. The protocol began with treatment of the hydrocephalus and confirmation of the diagnosis. Tumor surgery was performed either after conventional chemotherapy (eight patients) or after subsequent high-dose chemotherapy (HDCT; 11 patients). Two children with early leptomeningeal progression died before surgery. Craniospinal irradiation was applied to children older than 5 years of age, whereas younger children received local irradiation only. Hydrocephalus was present in 17 children and was treated with ventriculocisternostomy in 13 and shunt insertion in four. A biopsy procedure was performed with a stereotactic frame in 10 children, an open surgery was performed in four, an endoscope was used during the ventriculocisternostomy in three, and the diagnosis was made based on cerebrospinal fluid cytological analysis in two. The response rate to the first two courses of chemotherapy was 71% for the tumor and 59% for the metastases. The pathological analysis of the residue after chemotherapy showed true medulloblastomas in seven cases, complete neuroglial maturation in three cases, and a mixture of both in nine cases. Three-year progression-free survival was 37% and was significantly better in children older than 5 years of age. There was one death related to the HDCT. CONCLUSIONS: Preoperative chemotherapy is feasible and safe in children with high-risk medulloblastomas provided that the hydrocephalus can be treated at diagnosis. A larger study is warranted to ensure that the high response rate to adjuvant chemotherapy can lead to better surgical results and survival advantage. PMID- 16270683 TI - Meningioangiomatosis without neurofibromatosis: a clinical analysis. AB - OBJECT: Meningioangiomatosis is an uncommon clinical entity. This lesion has been reported at time of autopsy in patients with neurofibromatosis (NF) and in case reports of patients without NF Type 2 (NF2). The authors report a series of six patients with meningioangiomatosis who do not have NF2 and describe the clinical presentation, diagnosis of disease, and treatment. They also review the literature concerning this entity. METHODS: Six patients with meningioangiomatosis were treated at the authors' institutions from 1994 to 2001. The mean age of the patients was 10.7 years (range 5-14 years). All of the children presented with a seizure disorder. Surgery was performed in all children, and a gross-total resection was accomplished. All patients exhibited clinical improvement. At last follow up (mean 6.3 years) all patients are free of seizures and are not taking anticonvulsant medications. No signs of recurrence have been noted on imaging studies. CONCLUSIONS: The authors advocate a gross total resection of meningioangiomatosis for the treatment of seizure disorder in this population. PMID- 16270684 TI - Surgical strategies for approaching hypothalamic hamartomas causing gelastic seizures in the pediatric population: transventricular compared with skull base approaches. AB - OBJECT: The authors provide evidence that direct resection of hypothalamic hamartomas (HHs) can improve associated gelastic and nongelastic seizures. METHODS: Ten children younger than 17 years of age underwent resection of HHs (nine sessile and one pedunculated) that were causing refractory epilepsy. Lesions were approached from above transventricularly through a transcallosal anterior interforniceal approach in six cases, endoscopically through the foramen of Monro in one, and from below with a frontotemporal craniotomy including an orbitozygomatic osteotomy in three. Medical charts were reviewed retrospectively, and follow-up data were obtained through office records and phone calls. Follow up periods ranged between 12 and 84 months (mean 16.8 months). All patients in whom the approach was from above had sessile HHs. Five were free from seizures at follow up and two had a reduction in seizures of at least 95%. The transventricular route allowed excellent exposure and visualization of the local structures during resection. Among the three patients in whom the approach was from below, one became free of seizure after two procedures and one had a 75% reduction in epilepsy; the latter two had sessile HHs. The exposure was inadequate, and critical tissue borders were not readily apparent. Although the HH was adequately exposed and resected, the epilepsy persisted in the third patient, who had a pedunculated lesion. The overall rate of major permanent hypothalamic complications appeared to be slightly lower for the orbitozygomatic osteotomy group. CONCLUSIONS: Sessile lesions are best approached from above. Approaches from below adequately expose pedunculated hamartomas. The likelihood of curing seizures seems to be higher when lesions are approached from above rather than from below. PMID- 16270685 TI - Stereoelectroencephalography in the presurgical evaluation of children with drug resistant focal epilepsy. AB - OBJECT: The authors report on indications for, techniques of, and results of stereoelectroencephalography (stereo-EEG) monitoring in the presurgical evaluation of childhood drug-resistant focal epilepsy. METHODS: Thirty-five children (24 boys and 11 girls; mean age 8.8 +/- 3.9 years; mean age at seizure onset 2.1 +/- 2.2 years) underwent 37 tailored stereotactic placements of intracerebral, multilead electrodes for intensive monitoring of seizures (stereo EEG). Two children underwent exploration twice. Investigations prior to stereo EEG monitoring, including magnetic resonance imaging and video-EEG of the scalp, revealed variable patterns of localizing incoherence among clinical, electrophysiological, and anatomical findings in all cases. A total of 426 electrodes were implanted (mean 11.5 +/- 2.6 per procedure) according to individualized arrangements. The stereo-EEG monitoring enabled recording of 826 seizures (mean 22 +/- 45 per procedure) and mapping of critically functional areas. One untoward event consisted of intracranial breakage of an electrode. Resections of the epileptogenic zone were performed according to results of stereo-EEG recordings. They consisted of cortical removal in seven cases, removal of the lesion and epileptogenic cortex in 26 cases, and lesion removal in two cases. Transient postoperative neurological deficits were observed in 13 patients, and permanent anticipated visual field defects occurred in seven patients. According to the Engel postoperative classification of seizure outcome, 60% of cases were in Class I, 74% had significant improvement (Classes I-III), and 26% were unchanged (mean follow up 31.9 +/- 20.6 months). CONCLUSIONS: Stereoelectroencephalography monitoring is a helpful and well-tolerated technique for the definition of the epileptogenic zone in complex cases of childhood drug resistant epilepsy. It provides essential information for safe resections that should result in excellent outcomes in a considerable number of patients. PMID- 16270686 TI - Terminal myelocystoceles: a series of 17 cases. AB - OBJECT: A terminal myelocystocele is a rare form of spinal dysraphism in which the hydromyelic caudal spinal cord and the subarachnoid space are herniated through a posterior spina bifida. In their study of 17 cases, the authors attempt to develop treatment guidelines for patients suffering from this condition. METHODS: Seventeen patients (age range 2 months-5 years) underwent surgery during a 7-year period. Surprisingly, nine of 17 patients had no neurological deficits. Among these nine asymptomatic cases, three patients had a dermal sinus and two had a skin dimple, whereas one had congenital talipes equinovarus deformity (CTEV). Among the eight patients with neurological deficits, seven had lower-limb weakness (two had complete paraplegia), and one patient had urinary incontinence and constipation. Four patients each had a dermal sinus and CTEV; interestingly, one infant had an accessory phallus. Radiologically, eight patients had pure myelocystoceles, and a diagnosis of lipomyelocystocele was made in nine instances on the basis of magnetic resonance images. All patients underwent surgery. The meningocele sacs were excised, and the bands tethering the myelocystocele sac to the meningocele sacs were incised, with as little damage to the roots as possible. Two patients experienced postoperative CSF leakage (one required reexploration), and two others had subcutaneous CSF collection. No patient had deterioration of the neurological condition after surgery. CONCLUSIONS: The authors present the largest study of patients with terminal myelocystocele to date and emphasize that the possibility of this condition must be kept in mind when addressing all congenital lumbosacral masses. The authors assert that myelocystoceles and lipomyelocystoceles should be kept in the same category and recommend early surgery in all cases. PMID- 16270687 TI - Genetic considerations in nonsyndromic midline craniosynostoses: a study of twins and their families. AB - OBJECT: The origins of both sagittal synostosis (scaphocephaly) and metopic synostosis (trigonocephaly) remain unclear. Genetic and environmental factors probably play a role. Twin and family data of patients with these conditions are presented. METHODS: Records of 860 patients with scaphocephaly and 394 with trigonocephaly were examined for prevalence of twin birth and family history. Concordance rates of disease in monozygotic (MZ) and dizygotic (DZ) twins were compared. A family history of disease was present in 5.7% of children with scaphocephaly and in 6.8% with trigonocephaly. The frequency of twins was 4.2% in sagittal synostosis and 6.8% in metopic synostosis, which is higher than in the normal population. Twenty-eight of 38 twin pairs with a scaphocephalic proband were identified to be DZ and 10 were confirmed to be MZ (MZ/DZ ratio 0.36). Twenty of 27 twin pairs with trigonocephaly were DZ and seven were MZ (MZ/DZ ratio 0.35). Concordance rates were higher in the MZ group, 30% for scaphocephaly and 43% for trigonocephaly (chi2 = 14.4091, p = 0.0001). There was a strong prevalence of boys, with a frequency of 79.1% with scaphocephaly and 75.8% with trigonocephaly. Twinning was more frequent among affected boys in both groups. CONCLUSIONS: A midline craniosynostosis is more likely to develop in twins compared with singletons. A genetic component is supported by the higher risk in MZ twins. The presence of an environmental component is reinforced by the high rate of twinning, the normal MZ/DZ ratio, and a less than 100% concordance rate in MZ twins. PMID- 16270688 TI - Epidermal growth factor receptor gene amplification and expression in disseminated pediatric low-grade gliomas. AB - OBJECT: Pediatric low-grade gliomas (LGGs) are the largest group of central nervous system neoplasms in children. Although these tumors are generally benign, 5 to 10% of patients with pediatric LGGs present with leptomeningeal dissemination. The genetic and biological nature of these tumors is poorly understood. The authors looked for certain molecular abnormalities that may differentiate disseminated gliomas from the other pediatric LGGs. METHODS: Comparative genomic hybridization (CGH) was applied to 18 pediatric LGGs. Six cases featuring disseminated pediatric LGGs were compared with 12 control cases involving nondisseminated pediatric LGGs. Fluorescence in situ hybridization (FISH) analysis and immunohistochemical analysis were used to highlight further specific genetic targets. The CGH revealed multiple chromosomal abnormalities in five of six cases with disseminated gliomas and in six of 12 control cases. No correlation was found between the number of chromosomal abnormalities and dissemination status. Amplification of chromosome 7 was noted in four of six cases with disseminated gliomas as opposed to one of 12 control cases (p = 0.02). The FISH analysis revealed epidermal growth factor receptor (EGFR) amplification in one case negative to chromosome 7 amplification by CGH, raising the amplification cases to five of six (p = 0.0038). Immunohistochemical analysis for EGFR was positive in six of six cases and in two of 12 control cases (p = 0.0015). At the end of a mean follow-up period of 7.2 years, all patients with disseminated gliomas are alive with variable but slow disease progression. CONCLUSIONS: The high rate of EGFR gene amplification and protein expression in disseminated pediatric LGGs is intriguing and may have implications for our understanding of the role of EGFR in glioma genesis. Targeted therapies may be available for these children. Larger-scale studies are needed to establish further these findings. PMID- 16270689 TI - Desmoplastic fibroma of the pediatric skull. Report of three cases. AB - Desmoplastic fibromas are rare bone tumors that have been reported in the adult skull but rarely in that of children. Although desmoplastic fibromas of the pediatric skull are uncommon, their similarity to benign skull lesions and their locally aggressive nature make them an important part of the differential diagnosis. Local recurrence is common after curettage alone but complete resection appears to be curative. Close follow up of incompletely resected lesions is essential. The authors detail three cases of pediatric desmoplastic fibromas of the skull and discuss diagnosis and treatment. PMID- 16270690 TI - Measurement of flow of cerebrospinal fluid in shunts by transcutaneous thermal convection. Technical note. AB - With the goal of developing a practical method of performing noninvasive measurements of flow in cerebrospinal fluid (CSF) shunts, transcutaneous thermal convection CSF shunt flow measurement was investigated using dimensional analysis, numerical modeling, and bench testing. Using appropriate manufacturing practices and controls, a microcontroller-based device was designed, constructed, and clinically tested. Flow was detected in functioning shunts nine times in 10 attempts. One test failed due to postoperative edema, and subsequent testing was limited to patients who had not undergone shunt surgery within the previous 2 weeks. On the basis of these data and previous reports, 510(k) clearance was granted by the Food and Drug Administration for detection of flow in CSF shunts. Flow in CSF shunts can be detected noninvasively and cost effectively by using a simple thermal convection system. The positive and negative predictive values of the test are equal to or greater than those of brain imaging and radionuclide shunt studies. PMID- 16270691 TI - Improved survival in central nervous system aspergillosis: a series of immunocompromised children with leukemia undergoing stereotactic resection of aspergillomas. Report of four cases. AB - Central nervous system (CNS) aspergillosis remains a daunting diagnosis. This opportunistic mycosis historically carries a mortality rate approaching 100% in immunocompromised patients, with death ensuing within days after the onset of neurological symptoms. From their literature review, the authors concluded that children contracting CNS aspergillosis while undergoing systemic chemotherapy for leukemias represent a particularly unfortunate prognostic group. Antifungal medications prove ineffective for treating CNS aspergillosis in patients immunocompromised because of their chemotherapy regimens. In contrast, withholding chemotherapy to reverse immunosuppression, thereby improving the efficacy of antifungal medications, allows for progression of the primary leukemic disease. The authors present a series of four immunosuppressed patients whose course of treatment for leukemia was complicated by CNS Aspergillus sp. abscesses. Multiple cerebral fungal abscesses developed in two patients and a single cerebral abscess developed in two. All four patients underwent frameless stereotactic resection of the aspergilloma. All children later experienced resolution of their CNS infections and full neurological recovery. At 2- to 4 year follow ups, one patient has died of leukemia and the other three continue to thrive without evidence of recurrent aspergillosis. Given the grave natural history cited in the literature for this disease when medical treatment is instituted alone, the authors stress the crucial role of stereotactic neurosurgery for the intelligent treatment of immunocompromised children suspected of harboring a CNS aspergilloma abscesses. The authors propose that the goal for successful treatment in these patients should be gross-total resection of the abscess, its wall, and its capsule. PMID- 16270692 TI - Occipitocondylar hyperplasia: an unusual craniovertebral junction anomaly causing myelopathy. Case report. AB - The authors of this brief case report describe a previously unreported developmental variant of the craniovertebral junction. A 10-year-old girl who presented with cervical myelopathy was found to have cervicomedullary compression by hypertrophic occipital condyles. Decompression was achieved via a midline dorsal approach with no complications. The embryology of this area is briefly reviewed to attempt to explain the origin of this anomaly. PMID- 16270693 TI - Neurocutaneous melanosis. PMID- 16270694 TI - Evaluation of the In Touch training program for the management of alcohol and other drug use issues in schools. AB - In Touch is a professional training program designed to develop staff skills and support structures so as to enable schools to manage alcohol and other drug (AOD) matters in a coordinated manner that maximizes beneficial outcomes for at-risk students, while at the same time maintaining school discipline and community relationships. This study is an evaluation of the impact of the program on alcohol and other drug (AOD) related knowledge, attitudes and activity of participating school staff, and on AOD management practice in their schools. Data from 53 intervention participants and 21 controls were compared at pre- and post intervention. These data indicated a 46% increase in AOD knowledge among those who participated in In Touch training. Attitudes favorable to integrated, supportive management of AOD issues also increased significantly in this group, as did desirable practice. However, change in school practice was limited. Significantly more schools whose staff participated in In Touch training had a written drug policy at post-intervention, but schools' usual responses to AOD related incidents were substantially the same. These findings indicate that professional training on the management of AOD matters can change the understanding and practice of individual staff, but if school structures and practice are to be substantially influenced, a broader program is required. PMID- 16270696 TI - Current drug education policies in NCAA institutions: perceptions of head athletic trainers. AB - This study compared the perceptions of head athletic trainers (HATS) from NCAA member Divisions I, II, and III regarding current athletic department drug education policies in their institutions. A Web-based questionnaire collected responses from 353 HATS. Drug education programs focused more on providing information about the negative consequences of drugs than changing the attitudes and behaviors of students in relation to drug use. More Division I and II schools offered comprehensive ATOD education programs to their athletes compared to Division III schools; however, the majority of these programs were not scheduled on a regular basis. HATS in all divisions reported that the abuse of alcohol and other drugs by student athletes during and after athletic and social events was a significant concern and indicated the importance of scheduling drug education programs closer to such events. Majority of HATS did not view participating in the drug education programs as part of their professional duties; however, they expressed an interest to increase their involvement in these activities. PMID- 16270695 TI - An exploratory study examining the spatial dynamics of illicit drug availability and rates of drug use. AB - This study examines the spatial relationship between drug availability and rates of drug use in neighborhood areas. Responses from 16,083 individuals were analyzed at the zip code level (n = 158) and analyses were conducted separately for youth and adults using spatial regression techniques. The dependent variable is the percentage of respondents using drugs in the past year. Neighborhood drug availability (the major independent variable) was measured by the percentage of non-drug users who had been approached to purchase drugs. Data were obtained as part of the Fighting Back community evaluation. For youth (aged 12 to 18), drug sales in adjacent and surrounding areas were positively associated with self reported drug use in areas where youth were residents. For adults, drug sales within the neighborhood were negatively associated with drug use, while drug sales in immediately adjacent neighborhoods were positively related to self reports of drug use. Findings suggest that the areas where rates of drug users are greatest are not necessarily the same area where drugs are sold. Designing strategies to reduce the supply of drugs should receive input from city and regional planners and developers, as well as law enforcement and public health professionals. PMID- 16270697 TI - The relationship between perceived youth exposure to anti-smoking advertisements: how perceptions differ by race. AB - Data on self-reported perceived exposure to anti-smoking messages were collected from 1,608 high school students surveyed through the ASPIRE (A Smoking Prevention Interactive Experience) Program in Houston, Texas. Data collection took place between October 2002 and March 2003. Logistic regression identified that African Americans perceived significantly less exposure to anti-smoking advertisements via television (OR = .50, p < 0.05) and posters (OR = .61, p < 0.05) than whites. However, they had nearly twice as much perceived exposure to anti-smoking advertisements at movies (OR = 1.79, p < or = 0.05) and sporting events (OR = 2.1, p < or = 0.05) than their white counterparts. Hispanic youth perceived significantly less exposure to anti-smoking posters (OR = .51, p < or = 0.05) and significantly higher exposure to anti-smoking messages at sporting events (OR = 1.92, p < or = 0.05) and school programs (OR = 3.44, p < or = 0.05) compared to white youth. While the relationships tested in this study are exploratory, they provide initial evidence for an important nexus between race and communication channels that may impact perceived exposure to tobacco advertising. PMID- 16270698 TI - Trafficking in meth: an analysis of the differences between male and female dealers. AB - Male and female meth dealers exhibit numerous common characteristics and patterns. For example, both can be relatively heavy users and both have similar (long) criminal records. However, both groups show important distinctions in their drug dealing patterns. This exploratory study compares 34 male and 26 female meth dealers (N = 60) who were involved in the same criminal justice system of a single western state during the same time period. Among other things, the findings indicate that male dealers treat their trafficking activities more like a business, have more customers, make more money, and are more likely prepared for violence while dealing than females. In contrast, female dealers have more education, including graduate school, have been arrested less often and are more willing to try drug treatment than male dealers. PMID- 16270699 TI - Gender differences in predicting high-risk drinking among undergraduate students. AB - The purpose of this study was to examine gender differences in college students' high-risk drinking as measured by an estimated blood alcohol concentration (eBAC) based on gender, height, weight, self-reported number of drinks, and hours spent drinking. Using a developmental/contextual framework, high-risk drinking is conceptualized as a function of relevant individual characteristics, interpersonal factors, and contextual factors regularly mentioned in the college drinking literature. Individual characteristics include race, gender, and age; interpersonal characteristics include number of sexual partners and having experienced forced sexual contact. Finally, contextual factors include Greek membership, living off-campus, and perception of peer drinking behavior. This study is a secondary data analysis of 1,422 students at a large university in the Southeast. Data were gathered from a probability sample of students through a mail survey. A three-step hierarchical logistic regression analysis showed gender differences in the pathway for high-risk drinking. For men, high-risk drinking was predicted by a combination of individual characteristics and contextual factors. For women, interpersonal factors, along with individual characteristics and contextual factors, predicted high-risk drinking, highlighting the importance of understanding female sexual relationships and raising questions about women's risk-taking behavior. Implications for prevention and assessment are discussed. PMID- 16270700 TI - On caries prevalence and school-based fluoride programmes in Swedish adolescents. AB - Dental caries on approximal tooth surfaces in adolescents is still a problem in Sweden, as well as in many other industrialised countries. The aims of the present thesis were therefore: 1) to study whether caries prevalence is underestimated, 2) to evaluate the effect of cessation of fluoride mouth rinse (FMR) programme in schoolchildren with low caries prevalence, and 3) to reconsider the school as an arena for population-based fluoride (F) varnish and FMR programmes in order to minimise caries development in 13-16-year-olds. Paper I and II showed that the adolescents' own dentists consistently registered less caries compared to the recordings made by calibrated dentists from outside. Based on 420 16-year-olds at 12 different dental clinics in two neighbouring counties in Sweden, the differences were statistically significant at 10 of the 12 clinics when caries prevalence both was high in 1984 (Paper I) and somewhat lower in 1987 and 1990 (Paper II). About 80% of all approximal caries lesions were enamel lesions and are therefore not reported to the Swedish authorities, as only dentin lesions and fillings are currently included in the official caries data. The basis for Paper III was that many counties in Sweden abandoned school-based FMR programmes in the mid-late 1980s, as the official caries data revealed low caries prevalence among children. The cessation of FMR for 3 years for a group of 13-16 year-old adolescents (n=60) with low caries prevalence did not reveal any statistically significant differences in new caries lesions and fillings or in the progression of existing enamel lesions compared to a group of 13-16-year-olds (n=60) who continued to rinse for 3 years. Nor did a supplementary cross sectional study reveal any differences in caries prevalence among adolescents who had or had not taken part in FMR programmes for the last 3 years. In Papers IV and V, new models for school-based F treatment were evaluated in two 3-year randomised controlled trials (RCT studies). Fluoride varnish (Duraphat) treatment, carried out at school by specially trained dental nurses using a simple mobile unit, among 854 13-16-year-olds from low, medium and high caries risk areas showed that the control groups developed more caries than the varnish groups (Paper IV). The largest difference was found in the high-risk area. The prevented fraction (PF) with approximal enamel lesions as the diagnostic threshold was 69% in high, 66% in medium and 20% in low risk areas for F varnish applied twice a year at six-month intervals. Supervised school-based FMR among 788 13-16-year-olds with low to moderate caries risk (Paper V) showed that FMR on the first three and the last three school days during the school term, i.e. 12 rinses/year, had a PF of 59%. Enamel lesions constituted > 90% of the new approximal caries lesions in both Papers IV and V. The main conclusions from this thesis are: (1) that enamel caries lesions on approximal surfaces should be included in epidemiological caries data in order to show true caries prevalence, (2) that the cessation of weekly FMR for 3 years did not lead to a statistically significant increase in caries incidence in a relatively small group of adolescents with low caries prevalence, and (3) that the school should be reconsidered as an arena for new models for population-based F school programmes. Fluoride varnish treatment twice a year at six-month intervals in medium and high caries risk areas, as well as supervised FMR on the first and the last three school days during the school term in low and medium caries risk areas, are excellent caries prevention strategies for approximal surfaces in adolescents at the caries risk ages of 13-16 years, as a supplement to dental home care and preventive efforts at dental clinics. PMID- 16270701 TI - Jewish day-schooled adolescents' perceptions of parental and environmental support of physical activity. AB - Few studies of religiosity and physical activity (PA) correlates have been conducted; thus, a cross-sectional survey of 367 more and less religious Jewish adolescents' (M age = 12.8 years, SD = 0.9) perceptions of parental and environmental support of PA was conducted at religious day schools in two large American cities. Maternal Support, Environmental Support, and Paternal Non Support factors were verified through confirmatory factor analysis. Factor scores were analyzed through a 2 x 2 (gender x religiosity level) multivariate analysis of variance. A significant main effect for religiosity was obtained (F = 6.93, p < .001); however, a main effect for gender and an interaction effect were not. Post hoc discriminant analysis revealed Paternal Non-Support and Environmental Support factors as differentiating more and less religious adolescents. PMID- 16270702 TI - Comparison of accelerometer and pedometer measures of physical activity in boys and girls, ages 8-10 years. AB - Accelerometers record total physical activity and time spent at different intensities. Pedometers indicate total activity only. The aim of this study was to assess the relationship between pedometer counts and attainment of > or = 60 min of moderate activity. Thirty-four children, ages 8-10 years, wore a Tritrac accelerometer and Yamax pedometer. Published recommendations of steps per day were compared to attainment of > or = 60 min moderate activity. Boys who accumulated 13,000 steps.d(-1) and girls who accumulated 12,000 steps.d(-1) engaged in > or = 60 min moderate activity. However, 23% of boys and 15% of girls did not reach the pedometer thresholds but did engage in > or = 60 min moderate activity. In conclusion, these pedometer thresholds provide a reasonable estimation when assessment of physical activity intensity is not possible. PMID- 16270703 TI - Programming and reprogramming sequence timing following high and low contextual interference practice. AB - Individuals practiced two unique discrete sequence production tasks that differed in their relative time profile in either a blocked or random practice schedule. Each participant was subsequently administered a "precuing" protocol to examine the cost of initially compiling or modifying the plan for an upcoming movement's relative timing. The findings indicated that, in general, random practice facilitated the programming of the required movement timing, and this was accomplished while exhibiting greater accuracy in movement production. Participants exposed to random practice exhibited the greatest motor programming benefit, when a modification to an already prepared movement timing profile was required. When movement timing was only partially constructed prior to the imperative signal, the individuals who were trained in blocked and random practice formats accrued a similar cost to complete the programming process. These data provide additional support for the recent claim of Immink & Wright (2001) that at least some of the benefit from experience in a random as opposed to blocked training context can be localized to superior development and implementation of the motor programming process before executing the movement. PMID- 16270704 TI - Heat production and storage are positively correlated with measures of body size/composition and heart rate drift during vigorous running. AB - The purposes of this study were to determine the relationships between: (a) measures of body size/composition and heat production/storage, and (b) heat production/storage and heart rate (HR) drift during running at 95% of the velocity that elicited lactate threshold, which was determined for 20 healthy recreational male runners. Subsequently, changes in skin and tympanic temperatures associated with a vigorous 20-min run, HR, and VO2 data were recorded. It was found that heat production was significantly correlated with body mass (r = .687), lean mass (r = .749), and body surface area (BSA, r = .699). Heat storage was significantly correlated with body mass (r = .519), fat mass (r = .464), and BSA (r = .498). The percentage of produced heat stored was significantly correlated with body mass (r = .427), fat mass (r = .455), and BSA (r = .414). Regression analysis showed that the sum of body mass, percentage of body fat, BSA, lean mass, and fat mass accounted for 30% of the variability in heat storage. It was also found that HR drift was significantly correlated with heat storage (r = .383), percentage of produced heat stored (r = .433), and core temperature change (r = .450). It was concluded that heavier runners experienced greater heat production, heat storage, and core temperature increases than lighter runners during vigorous running. PMID- 16270705 TI - The career planning, athletic identity, and student role identity of intercollegiate student athletes. AB - The purpose of this study was to examine the career planning of university student athletes and relationships between their career planning and athletic and student role identities. Two retrospective in-depth interviews were held with four male and four female university student athletes. Participants entered university with vague or nonexistent career objectives and invested heavily in their athletic roles. In the latter years of their college career, the participants discarded their sport career ambitions and allowed the student role to become more prominent in their identity hierarchies. The current findings support Brown and Hartley's (1998) suggestion that student athletes may invest in both the athlete and student role identities simultaneously and that investing in the latter may permit the exploration of nonsport career options. PMID- 16270706 TI - The parent-coach/child-athlete relationship in youth sport: cordial, contentious, or conundrum? AB - The roles of coach and parent are often synonymous in youth sport, but little data-based research has been conducted on the parent-coach/child-athlete relationship. Six boys in U-12 competitive soccer were interviewed regarding positive and negative aspects about playing for their father-coach. Similar questions were posed to father-coaches and two teammates. Inductive content analysis indicated that, among the benefits, sons identified perks, praise, technical instruction, understanding of ability level, insider information, involvement in decision making, special attention, quality time, and motivation. Costs of being coached by one's father included negative emotional responses, pressure/expectations, conflict, lack of understanding/empathy, criticism for mistakes, and unfair behavior. For father-coaches, positive themes included taking pride in son's achievements, reason for coaching, positive social interactions, opportunity to teach skills and values, enjoying coaching son, and quality time. Negatives included inability to separate parent-child from coach player role, placing greater expectations and pressure on son, and showing differential attention toward son. While teammates perceived some favoritism by the parent-coach, they cited mostly positive instructional experiences. Results are discussed within motivational theories that highlight the influence of significant adults on children's psychosocial development in the physical domain. PMID- 16270707 TI - Deliberate imagery practice: the reliability of using a retrospective recall methodology. AB - This study examined the reliability of a retrospective recall methodology for providing evidence of deliberate imagery practice. A secondary purpose was to determine which imagery activities constituted the sport-specific definition of deliberate practice (Starkes, Deakin, Allard, Hodges, & Hayes, 1996). Ninety three Canadian athletes from one of three different competitive levels (regional, provincial, and national) completed the Deliberate Imagery Practice Recall Questionnaire, which was specifically designed for the present study. The athletes also completed a 1-week imagery diary to assess their use of 14 different imagery activities. The results of the study indicated that the athletes were able to reliably estimate their use of imagery over a short recall interval. Four imagery activities were also determined to fulfill the sport specific definition of deliberate practice. PMID- 16270708 TI - Comparability and reliability of paper- and computer-based measures of psychosocial constructs for adolescent physical activity and sedentary behaviors. AB - This study assessed the comparability and reliability of paper-based and computer based administration of psychosocial construct measures related to adolescents engaging in physical activity and sedentary behaviors. Adolescents (n = 76; 55% girls, 49% Hispanic, 18% non-Hispanic-white; M age = 13 years) completed measures of behavior change strategies, self-efficacy, decisional balance, family and peer influences, enjoyment, activity choices, and environmental influences. Overall, the adolescents provided equivalent responses on paper and computer formats. Reliability estimates were generally good for the multiple-item constructs, but single-item measures tended to demonstrate low reliability. Average scale internal consistency was significantly higher for measures administered with the computer format compared to the paper format, but test-rest reliability estimates were not significantly different between formats. PMID- 16270709 TI - Volunteer youth sport coaches' perspectives of coaching education/certification and parental codes of conduct. AB - The vast majority of youth sport programs in the United States relies primarily on parent volunteers to serve as coaches. Unfortunately, most of these volunteer coaches have not received formal training to prepare them adequately for the role of youth sport coach. To exacerbate the issue, according to the popular media, parents and other adults can commit belligerent and even violent acts around, and often resulting from, poorly managed youth sport events. Although some efforts have been made to standardize curricula, provide training for coaches, and contain or prevent inappropriate parent behaviors, few efforts have been directed at investigating the self-described needs and concerns of the coaches from their perspectives. The purpose of the current study was to investigate the concerns and issues of youth sport coaches related to coaching and parental education. Five focus group interviews with 25 volunteer youth sport coaches were conducted to investigate these issues. Results were organized around four higher order themes that emerged from inductive content analyses: (a) coaching education content areas of need, (b) barriers and problems of offering coaching education, (c) coaching education format recommendations, and (d) efficacy of parental codes of conduct. Results were discussed in terms of the potential impact administrators, coaches, and parents could have in implementing formal coaching education programs and developing their coaching education practices. PMID- 16270710 TI - Test-retest stability of the Task and Ego Orientation Questionnaire. AB - Establishing stability, defined as observing minimal measurement error in a test retest assessment, is vital to validating psychometric tools. Correlational methods, such as Pearson product-moment, intraclass, and kappa are tests of association or consistency, whereas stability or reproducibility (regarded here as synonymous) assesses the agreement between test-retest scores. Indexes of reproducibility using the Task and Ego Orientation in Sport Questionnaire (TEOSQ; Duda & Nicholls, 1992) were investigated using correlational (Pearson product moment, intraclass, and kappa) methods, repeated measures multivariate analysis of variance, and calculating the proportion of agreement within a referent value of +/-1 as suggested by Nevill, Lane, Kilgour, Bowes, and Whyte (2001). Two hundred thirteen soccer players completed the TEOSQ on two occasions, 1 week apart. Correlation analyses indicated a stronger test-retest correlation for the Ego subscale than the Task subscale. Multivariate analysis of variance indicated stability for ego items but with significant increases in four task items. The proportion of test-retest agreement scores indicated that all ego items reported relatively poor stability statistics with test-retest scores within a range of +/ 1, ranging from 82.7-86.9%. By contrast, all task items showed test-retest difference scores ranging from 92.5-99%, although further analysis indicated that four task subscale items increased significantly. Findings illustrated that correlational methods (Pearson product-moment, intraclass, and kappa) are influenced by the range in scores, and calculating the proportion of agreement of test-retest differences with a referent value of +/-1 could provide additional insight into the stability of the questionnaire. It is suggested that the item-by item proportion of agreement method proposed by Nevill et al. (2001) should be used to supplement existing methods and could be especially helpful in identifying rogue items in the initial stages of psychometric questionnaire validation. PMID- 16270711 TI - The effect of venue and wind on the distance of a hammer throw. PMID- 16270712 TI - Active transportation to school: findings from a national survey. PMID- 16270713 TI - Comparing self-reported versus objectively measured physical activity behavior: a preliminary investigation of older Filipino American women. PMID- 16270714 TI - Validation evidence for the Netherlands physical activity questionnaire for young children: the Iowa bone development study. PMID- 16270715 TI - Motivation and burnout in professional rugby players. PMID- 16270716 TI - Biomarkers and childhood asthma: improving control today and tomorrow. AB - Although we aim to normalize the lives of children with asthma by controlling their day and night symptoms and preventing exacerbations and morbidity, optimal childhood asthma management may result when the assessment and monitoring of asthma includes measured biomarkers--meaning objective, biological measures of lung dysfunction and inflammation. Precedence for such an approach to optimizing disease control and outcomes can be appreciated in comparing asthma with insulin dependent diabetes mellitus (IDDM) management in children. Optimal management of these chronic conditions shares the fundamental goals to eliminate day and night symptoms and prevent exacerbations and morbidity. However, IDDM management focuses primarily on peripheral blood biomarkers of tight control (i.e., daily serum glucose levels) and predictors of long-term morbidity (i.e., hemoglobin A1C, or hemoglobin "remodeling" due to chronically poor control of glucose) for optimal assessment and monitoring and to best achieve these clinical objectives (Alemzadeh R, et al. Diabetes mellitus in children. In Nelson Textbook of Pediatrics, 17th ed. Behrman RE, Kliegman RM, and Jenson HB (Eds). Philadelphia: W.B. Saunders Co., 1947-1972, 2004). The improved outcomes in IDDM have resulted primarily from the progress to a biomarker-based assessment to achieve tight, optimal control and not, presently, as a dramatic change in therapy. The progress in IDDM management provides a compelling paradigm to consider for improving childhood asthma management. Indeed, the time is good to not only consider some newly available biomarkers, but also to reconsider some biomarkers of lung dysfunction, inflammation, and atopy that could be broadly used today. This article reconsiders the use of current and emerging measures of lung dysfunction, inflammation, and atopy in assessing tight control and long-term risk. Concluding emphasis will be placed on what can be implemented today. PMID- 16270717 TI - The allergist and the workplace: occupational asthma and rhinitis. AB - Occupational respiratory disease is a significant problem and appears to be on the rise. The allergist should make every effort to become familiar with the workplace environment and appreciate the many causes of occupational rhinitis and asthma. In addition to prescribing appropriate medications, the clinician should be the patients total health advocate in terms of encouraging changes in the working environment to benefit the patient. Inattention to this may result, particularly in the case of occupational asthma, in progressive respiratory deterioration. PMID- 16270718 TI - Critical analysis of studies concerning reports of respiratory sensitization to certain wood dusts. AB - Studies have been published reporting that exposures to certain wood dusts are sensitizing, resulting in respiratory symptoms in susceptible individuals. Many of the publications in this field are case reports that collectively have a number of important shortcomings. Illuminating these should further our understanding of whether respiratory sensitization results from occupational exposure to particular wood dusts. The aim of this study was to critically review and understand the evidence to date regarding reported respiratory sensitization in connection with wood dusts from oak, beech, pine, ash, and western red cedar. Publications dealing with these commercially important woods in North America have been selected from the Pubmed/Medline database (1966 to the present) using the key word, wood dust. These articles, along with supporting references on procedures and techniques, are reviewed according to the strengths and weaknesses of evidence and conclusions presented. Evidence from skin testing, specific immunoglobulin E measurements, and basophil histamine release tests suggests that reported symptoms are not likely to be immunologically derived. Because of methodological problems, challenge tests with specific wood dusts do not support the conclusion that reactions to certain wood dusts are specific. Experiments with nonspecific bronchoconstrictive agents indicate that a number of study subjects possess hyperresponsive airways. Thus, select individuals can demonstrate various respiratory symptoms in the woodworking industry, but any specificity or direct cause is currently unproved. Current studies do not support that exposure to wood dusts from a number of common North American wood species causes immunologic sensitization in woodworkers. Rather, symptoms reported in some studies of exposed workers seem to follow the paradigm for nonspecific respiratory responses in individuals with hyperresponsive airways. PMID- 16270719 TI - The effect of intranasal steroid budesonide on the congestion-related sleep disturbance and daytime somnolence in patients with perennial allergic rhinitis. AB - Patients with perennial allergic rhinitis (PAR) often present with nasal congestion, poor sleep, daytime fatigue, and daytime somnolence. Pharmacologic therapy that reduces nasal congestion should improve the PAR patients' sleep quality and reduce daytime somnolence and fatigue. Our hypothesis is that intranasal steroid budesonide (BUD), an effective topical anti-inflammatory agent, will reduce nasal congestion and improve the patients' quality of life. The objective of this study was to determine whether topical steroid BUD improves sleep, daytime somnolence, and fatigue in patients with PAR. Twenty-six subjects were enrolled in a double-blind, placebo-controlled, crossover study using Balaam's design. Patients were treated with intranasal steroid spray BUD or placebo. The Epworth Sleepiness Scale, daily diary, and questionnaires were used as tools for subjective data analysis, which focused on nasal symptoms, sleep quality, daytime somnolence, and fatigue. The results were summarized and compared by PROC MIXED in SAS. The daily diary data showed significant improvement in self-reported nasal congestion (p = 0.04) and daytime sleepiness (p = 0.01) and a trend in reduction of daytime fatigue (p = 0.08) in the BUD group compared with the placebo group. The sleep measures showed statistically significant improvement in total sleep measures score (p = 0.04), "sleep compared with absolute" (p = 0.01), and "refreshing and restorative" sleep (p = 0.04) in the active group. Nasal corticosteroid BUD is effective in reducing nasal congestion, daytime somnolence, and daytime fatigue, and improving sleep quality in PAR. PMID- 16270720 TI - Comparative efficacy of cetirizine and fexofenadine for seasonal allergic rhinitis, 5-12 hours postdose, in the environmental exposure unit. AB - In a previous study, cetirizine and fexofenadine similarly relieved seasonal allergic rhinitis symptoms in the first 5 hours, but cetirizine was more effective at 21-24 hours postdose. This randomized, double-blind, placebo controlled study compared the response to treatment between 5 and 12 hours. Eligible ragweed allergic subjects were exposed to pollen in the Environmental Exposure Unit and randomized (n = 599) to a single dose of cetirizine, 10 mg; fexofenadine, 180 mg; or placebo (2.5:2.5:1). The primary efficacy end point was the change from baseline in total symptom severity complex (TSSC) score at 12 hours postdose. TSSC score was the sum of self-rated scores (0 = absent to 3 = severe) for runny nose, sneezing, itchy nose/palate/throat, and itchy/watery eyes, recorded half-hourly. Mean baseline TSSC scores were similar: 9.2, cetirizine and fexofenadine; 8.9, placebo. Reductions in TSSC scores from baseline were 4.3 at 12 hours and 5.0 overall (i.e., average over 5-12 hours postdose) for cetirizine and 3.4 and 4.4, respectively, for fexofenadine. Cetirizine produced a 26% greater reduction in TSSC at 12 hours (p = 0.001) and 14% greater reduction in TSSC overall (p = 0.006) compared with fexofenadine. Cetirizine and fexofenadine reduced TSSC scores (p < 0.001) and individual symptoms (p < 0.05) more than placebo. However, cetirizine was more effective than fexofenadine (p < 0.05) for runny nose and sneezing (12 hours and overall), itchy/watery eyes (12 hours), and itchy nose/throat/palate (overall). Incidence of treatment-emergent adverse events and somnolence were similar among groups: cetirizine, 25.3 and 0.8%, respectively; fexofenadine, 29.6 and 0%, respectively; placebo, 35.0 and 0%, respectively. In conclusion, cetirizine produced greater relief of seasonal allergic rhinitis symptoms than fexofenadine at 12 hours postdose and over the 5- to 12-hour postdose period. PMID- 16270721 TI - Montelukast versus formoterol as second-line therapy in asthmatic children exposed to relevant allergens. AB - In sensitive asthmatic children, the exposure to relevant allergens causes a deterioration of lung function and symptoms associated with an increase of inflammatory indices. The aim of this single-blind randomized add-on study was to compare the effects of montelukast or formoterol added to low-dose budesonide in asthmatic allergic children exposed to relevant allergens. Twenty children (5 female subjects and 15 male subjects, aged 6-12 years) were enrolled. Lung function and airway inflammatory indices (exhaled nitric oxide [eNO] and sputum eosinophils) were evaluated at T0 when children were not exposed to relevant allergens and at T1 after 15 days of natural effective allergen exposure. At T1, pulmonary function tests and sputum eosinophils remained stable in both of the groups, without significant differences in comparison with T0 at baseline. Furthermore, formoterol plus budesonide was effective in preventing the expected increase in eNO from 26.46 +/- 2.62 ppb at T0 to 29.33 +/- 9.28 ppb at T1 (not significant). However, in the group receiving montelukast plus budesonide, there was a significant decrease of eNO from baseline (30.78 +/- 6.87 ppb) to T1 (18.17 +/- 6.60 ppb) (p < .05). In allergic asthmatic children, the use of montelukast or formoterol combined with budesonide could offer a durable protective effect on symptoms, lung function, and inflammatory indices. PMID- 16270723 TI - The effect of fexofenadine on expression of intercellular adhesion molecule 1 and induction of apoptosis on peripheral eosinophils. AB - The role of eosinophils in the pathogenesis of allergic disorders has been established by several studies. Recently, it has been suggested that second generation antihistamines, widely used to relieve allergic symptoms, may have anti-inflammatory effects. To assess the possible anti-inflammatory activity of fexofenadine, a selective H1-receptor antagonist, we evaluated its capacity to modulate the expression of adhesion molecules leukocyte function-associated antigen (LFA) 1 and intracellular adhesion molecule (ICAM) 1 on eosinophil surface and to induce apoptosis of eosinophils. To analyze the expression of adhesion molecules, eosinophils from healthy donors were cultured in the presence of interferon gamma and tumor necrosis factor alpha with various concentrations of fexofenadine, incubated with monoclonal antibodies anti-ICAM-1 and LFA-1 and then analyzed by flow cytometry. To evaluate apoptosis of eosinophils, cells stimulated with interleukin-5, in the presence of different concentrations of fexofenadine, have been incubated with a phosphatidylserine-binding protein (annexin V) fluorescein isothiocyanate conjugated and then analyzed by flow cytometry. Apoptosis was evaluated as a percentage of annexin V+ cells. In this study, fexofenadine did not cause any significant changes in the expression of LFA-1 but was shown to be able to inhibit the expression of ICAM-1 at concentrations between 10(-3) and 10(-4) M. Moreover, concentrations of fexofenadine from 10(-3) to 6 x 10(-4) M induced a significant increment in the percentage of apoptotic cells. Our findings indicate the possibility of obtaining relevant anti-inflammatory pharmacologic effects, other than antihistamine activity, by fexofenadine, such as inhibition of ICAM-1 expression and induction of eosinophil apoptosis. PMID- 16270722 TI - Evaluation of theophylline or pranlukast, a cysteinyl leukotriene receptor 1 antagonist, as add-on therapy in uncontrolled asthmatic patients with a medium dose of inhaled corticosteroids. AB - A few studies compared the additional effects of oral controller medicines on pulmonary function in asthmatic patients on a moderate dose of inhaled steroids. The aim of this study was to compare the additional effects of two oral asthma controllers, a leukotriene receptor antagonist and a sustained released theophylline (Theo), with a moderate dose of inhaled steroid on peak expiratory flow (PEF) and asthma-related symptoms. A total of 67 adult asthmatic patients with PEF < 80% predicted during a 2-week run-in period with 800 microg/day of beclomethasone dipropionate were randomized to receive either pranlukast, 450 mg/day (n = 33), or sustained released Theo, 200 mg/day (n = 34), for 4 weeks. Pranlukast and Theo did not significantly alter the symptom scores, use of rescue beta2-agonist, and daily PEF variability. However, both agents significantly increased both morning and evening PEF compared with the run-in periods. The effects of both medications were comparable. For asthmatic patients even on a moderate dose of inhaled steroids, the addition of either leukotriene receptor antagonist or sustained released Theo does not improve asthma-related symptoms but significantly and equally increases PEF. PMID- 16270724 TI - Modulation of toll-like receptor 7 and LL-37 expression in colon and breast epithelial cells by human beta-defensin-2. AB - Breast-feeding decreases maternal breast cancer risk. Breast-fed infants have fewer infections and inflammatory-allergic diseases. We recently found inducible antimicrobial and immunomodulatory protein human beta3-defensin 2 (HBD-2) in significant amounts in human milk. We investigated if HBD-2 could contribute to benefits of breast-feeding for the mother and the child by immunomodulating effects on breast and gut epithelial cells. Human CaCo-2 colon and MCF-7 breast cell lines were cultured for 16-48 hours in RPMI 1640 5% fetal calf serum with and without HBD-2 at 0.1, 0.5, and 1.0 microg/mL. RNA was extracted and reverse transcription polymerase chain reaction (RT-PCR) and gel electrophoresis for toll like receptor pathway members, antimicrobial peptides, and cytokines/receptors was performed. Primers were designed with www.ncbi.nlm.nih.gov and www.broad. mit.edu/cgibin/primer/primer3 www.cgi. Based on RT-PCR results, cells were stained by immunohistochemistry using anti-toll-like receptor (TLR)-7 and anti LL37 antibodies and DAKO EnVision Plus kits. Supernatants were analyzed for interleukin (IL)-8 and liver and activation-regulated chemokine (LARC) using enzyme-linked immunosorbent assay. In CaCo-2, messenger RNA (mRNA) for TLR-7, IL 1R-associated kinase, alpha-defensins (human neutrophil peptides 1-3), and IL-8 were down-regulated; cathelicidin/LL37 and NFkappaBp65 were up-regulated. LARC mRNA and protein were detected after 48 hours. TLR-7 protein, LARC, and IL-8 decreased with HBD-2; LL-37 protein greatly increased. In MCF-7, mRNA for LL37, inhibitor of kappaBalpha, NFkappaBp65, Tollip, MyD88, IL-1R-associated kinase, and TLR-7 were up-regulated. LARC mRNA was turned off. TLR-7 protein was induced. LARC was not detected. IL-8 was barely detectable with or without HBD-2. beta Defensins 1 and 2; alpha-defensins 5 and 6; TLRs 1, 2, 3, 4, 5, 6, 8, 9, and 10; nucleotide binding oligomerization domain protein-2, and CCR6 mRNA were unaffected. HBD-2 profoundly alters the innate immune response of breast and intestinal epithelial cells. PMID- 16270725 TI - Demographics and long-term follow-up in a Veterans Affairs Allergy/Immunology Center: a 10-year analysis. AB - Between 1985 and 1992, patients were evaluated as new outpatient consultations in the Allergy/Immunology Center at the Veterans Affairs Greater Los Angeles Health Care System (VAGLAHS). Data collected included age, gender, ethnicity, and diagnosis. After 10 years, patient follow-up status was determined and classified into five categories: gender, ethnic distribution, age distribution, disorders seen, and follow-up pattern. A total of 1116 patients were evaluated. The gender of our population was 7.8% women and 92.2% men. The ethnic distribution was 59.5% white, 32.2% black, 6.5% Hispanic, and 1.9% other. Neither age nor ethnic distribution was significantly different from the general veterans affairs population. Age of patients ranged from 20 to 90 years old. The largest peak for age at initial presentation was 60 years. The three most common disorders seen in the clinic were rhinitis (36.6%), asthma (24.5%), and sinusitis (12.3%). The 10 year follow-up pattern of patients revealed that 6% were seen in the past year, 6.2% of patients were seen longer than 1 year ago but within the 5 past years, 29% of patients who were still seen at VAGLAHS but were no longer patients of the allergy clinic, 32.7% of patients who were no longer seen at VAGLAHS, and 26.2% died. Women and patients who were 50-60 years old were more likely to follow-up. There was no difference in follow-up visits among ethnic groups. PMID- 16270726 TI - Prompt remediation of water intrusion corrects the resultant mold contamination in a home. AB - More patients are turning to their allergists with symptoms compatible with allergic rhinitis, allergic sinusitis, and/or bronchial asthma after exposure to mold-contaminated indoor environments. These patients often seek guidance from their allergists in the remediation of the contaminated home or office. The aim of this study was to determine baseline mold spore counts for noncontaminated homes and report a successful mold remediation in one mold-contaminated home. Indoor air quality was tested using volumetric spore counts in 50 homes where homeowners reported no mold-related health problems and in one mold-contaminated home that was remediated. The health of the occupant of the mold-contaminated home also was assessed. Indoor volumetric mold spore counts ranged from 300 to 1200 spores/m3 in the baseline homes. For the successful remediation, the mold counts started at 300 spores/m3, increased to 2800 spores/m3 at the height of the mold contamination, and then fell to 800 spores/m3 after remediation. The occupant's allergic symptoms ceased on complete remediation of the home. Indoor volumetric mold counts taken with the Allergenco MK-3 can reveal a potential indoor mold contamination, with counts above 1000 spores/m3 suggesting indoor mold contamination. Once the presence of indoor mold growth is found, a prompt and thorough remediation can bring mold levels back to near-baseline level and minimize negative health effects for occupants. PMID- 16270727 TI - Dyspnea, wheezing, and airways obstruction: is it asthma? AB - Dyspnea, wheezing, and decreased FEV1 with bronchodilator response are characteristic of asthma. However, when standard asthma therapy fails, a broad differential must be considered to avoid a catastrophic outcome. This article presents a case report of a 48-year-old Filipino woman, who was referred for evaluation of cough, dyspnea and wheezy respiration, changes in voice quality, nasal and palatal pruritus, and postnasal drainage. She was found to have mold spore hypersensitivity and abnormal spirometry with an obstructive pattern and a 15% reversibility postnebulized albuterol. An initial diagnosis of allergic rhinitis and adult-onset asthma was made, and therapy was initiated which included: salmeterol, budesonide, montelukast, and pirbuterol. Her symptoms persisted and rabeprazole was added to treat possible laryngopharyngeal reflux. Repeat spirometry demonstrated worsening obstruction. There was no improvement with systemic corticosteroids. High-resolution computed tomography of the chest demonstrated a left paratracheal mass, obstructing 60% of the airway. Bronchoscopy revealed a tumor 4-5 cm below the vocal cords with the appearance of adenoid cystic carcinoma, which was confirmed by pathology. All symptoms resolved and spirometry normalized with resection of mass and radiation therapy. Adenoid cystic carcinoma (ACC) is an uncommon form of malignant neoplasm that arises from salivary glands. Tracheobronchial ACC typically presents with symptoms of cough, dyspnea, and hoarseness. ACC has a relatively indolent course. Standard therapy is surgical resection often followed by radiotherapy. In patients who fail conventional therapies for asthma, it is important to consider other diagnoses to avoid fatal outcomes. PMID- 16270728 TI - Asthma in pregnancy. AB - Asthma, one of the most common serious medical problems to complicate pregnancy, affects 3-8% of pregnancies in the United States. The goals of therapy in the pregnant asthmatic patient do not differ from those in non-pregnant patients. Inhaled corticosteroids (ICS) are preferred in the management of all levels of persistent asthma in pregnant patients, because these agents have been shown to reduce asthma exacerbations during pregnancy. Asthma in pregnancy is often undertreated due to physician and patient concerns over the effects of asthma medications on the fetus. However, undertreatment leads to loss of asthma control and increases in maternal morbidity, perinatal mortality, preeclampsia, preterm birth, and low birth weight infants. Recent prospective clinical cohort studies with active asthma management by NAEPP guidelines show no evidence of increased maternal or fetal morbidity or mortality. Therefore, it is critical for the mother to understand that failure to control asthma during pregnancy may lead to poor outcomes. A case study follows to highlight clinical pearls and pitfalls in the management of asthma in the pregnant patient. PMID- 16270729 TI - Joint Council of Allergy, Asthma, and Immunology News: focus on Practice Management, Part 2. PMID- 16270730 TI - Renal community works together to save lives. PMID- 16270731 TI - People working together to provide help and healing. PMID- 16270732 TI - Computer models enlisted in diabetes prevention. PMID- 16270733 TI - How method I and method II payments differ for home dialysis supplies and services. PMID- 16270734 TI - American Kidney Fund reaches out to minority communities at risk for CKD. PMID- 16270735 TI - How reimbursement affects medical practice. PMID- 16270736 TI - An analysis of socialized renal medicine. PMID- 16270737 TI - An analysis of socialized renal medicine. PMID- 16270738 TI - Paying fairly for good outcomes: which formula works? AB - I see the single-based payment systems moving more toward the middle, toward an American-style system using competitive bidding with outcomes-based focus. The American system is moving in the opposite direction despite knowledge of the flaws of a single-payment system. Modernizing our payment system by desegregating Medicare Part A and Part B, applying information technology, and value-based reimbursement along with a bold move toward an HSA methodology could empower patients and physicians, reinvigorate our system with cost savings, and move us in the direction that all industries in a free market system should have the freedom to move. PMID- 16270739 TI - How to improve communication in dialysis units: a survey of family focus readers. AB - Future study of communication between dialysis patients and health care providers could not only aid in understanding the survey findings but also advance emphasis on those areas of care needing to be enhanced. By raising awareness about communication in the dialysis unit, it is hoped that health care professionals will embrace the need to encourage patients to be more involved in the educational and treatment process, enabling them to become active partners in their health care. PMID- 16270740 TI - How renal professionals can deal with compassion fatigue. PMID- 16270741 TI - Understanding central venous catheter use among ESRD patients: a network 1 study. PMID- 16270742 TI - Has the MMA helped or hurt peritoneal dialysis programs? PMID- 16270743 TI - The avian flu: how scared should we be? PMID- 16270744 TI - Screening the priests. PMID- 16270745 TI - Aging naturally. PMID- 16270746 TI - Dr. Andrew Weil's wellness diet. PMID- 16270747 TI - Lessons from the 1918 flu. PMID- 16270748 TI - Topogenesis of peroxisomal proteins does not require a functional cytoplasm-to vacuole transport. AB - Folded and even oligomeric proteins can be imported from the cytosol into vacuoles and into peroxisomes. Pro-aminopeptidase I (prAPI) oligomerizes into a dodecamer and is imported into the vacuole via the cytoplasm-to-vacuole transport (cvt) pathway. How peroxisomes accommodate folded proteins is completely unknown. Peroxisome biogenesis and cvt do not only share the import of folded protein complexes but also show mechanistic parallels such as the employment of ubiquitin conjugation systems. In search for a genetic overlap, selected cvt and autophagocytosis (atg) mutants were tested for defects in peroxisome biogenesis. Most of the mutants did not exhibit a mislocalization of peroxisomal matrix proteins to the cytosol which would be typical of a defect in the peroxisome biogenesis. However, two mutants, deltaatg14 and deltacvt4/vam6, displayed a general growth defect and deltacvt8/vps41 showed cytosolic mislocalization not only of peroxisomal but also of mitochondrial proteins, indicating a more general defect in organelle biogenesis. Our data do not provide evidence for a genetic overlap of the import pathway for peroxisomal proteins and the cvt pathway. PMID- 16270749 TI - Phospho-eNOS Ser-114 in human mesenchymal stem cells: constitutive phosphorylation, nuclear localization and upregulation during mitosis. AB - Activity of endothelial nitric oxide synthase (eNOS) is modulated by protein protein interaction and phosphorylation at specific serine or threonine residues. Using immunofluorescence analysis we show here that proliferating mesenchymal stem cells (MSCs) derived from human bone marrow exhibit cytosolic and pronounced nuclear localization of eNOS. Examination of phosphorylated eNOS subspecies revealed that eNOS phosphorylated at Ser-114 is heavily enriched in the nucleus, whereas eNOS phosphorylated at Ser-1177 is localized at filamentous structures in the cytosol that are abundant in the perinuclear region. Phosphorylation of eNOS at Ser-114 but not at Ser-1177 was strongly increased in cells shortly before mitosis and decreased to normal level after completed cell division. Double immunofluorescence analysis revealed that subcellular localization of 8 hydroxyguanosine immunoreactivity was overlapping with eNOS phosphorylated at Ser 114 in human MSCs providing evidence that phosphorylation at this residue is linked to the generation of superoxide anions. As expected there was only a weak colocalization between eNOS phosphorylated at Ser-1177 and caveolin-1. Different from many other cell systems, human MSCs accumulate eNOS in the nucleus without an acute stimulus. eNOS constitutively phosphorylated at distinct amino acid residues is targeted to different subcellular compartments pointing to an important role of specific phosphorylation events in the life cycle of proliferating human MSCs. PMID- 16270750 TI - Increased association with detergent-resistant membranes/lipid rafts of apically targeted mutants of the interleukin-6 receptor gp80. AB - Interleukin (IL)-6 is an important cytokine in inflammatory processes, differentiation and growth. The IL-6 receptor complex comprises the specific IL-6 receptor (gp80) and two molecules of the signal tranducing component gp130 which transduces the signal into the nucleus via the Jak-STAT pathway. Both, gp80 and gp130 are sorted preferentially to the basolateral membrane of polarised Madin Darby canine kidney (MDCK) cells. Previously, we have shown that gp130 partially localises to detergent-resistant membranes (DRMs)/lipid rafts and that lipid raft integrity is crucial for signalling to occur. Here we now demonstrate that wild type gp80 is associated with DRMs only to a minor extent. However, gp80 mutants which lack parts of the cytoplasmic domain and therefore are more apically expressed than the wild type show an increased affinity for the liquid-ordered membrane domain. Studies with non-polarised MDCK cells suggest that the lipid raft association of the different mutants of gp80 precedes the establishment of cell polarity. Our findings suggest that lipid rafts play a role in the sorting of apically targeted gp80. PMID- 16270751 TI - Modulation of chondrocyte adhesion to collagen by echistatin. AB - Primary chondrocytes from quail embryo epiphysis (quail epiphyseal chondrocytes, QEC) can grow either in suspension or in monolayer. In this study, the adhesion of QEC to collagen II was used as a model to study the regulation of the ligand binding activity of integrin receptors that allows these cells to undergo a rapid transition from suspension to an adherent state. Preincubation of suspension QEC (QECSP) with the disintegrin echistatin increased by 40% their adhesion to collagen II. An inverse relationship between immobilized collagen density and echistatin-induced increase of chondrocyte adhesion was observed, thus suggesting that the disintegrin acts by increasing the ligand-binding affinity of collagen receptor(s). Further, echistatin activity does not appear to depend upon a direct binding of the disintegrin to collagen receptor(s). In fact, immobilized anti beta1 antibodies, but not immobilized echistatin, served as effective binding sites for QECSP. Echistatin failed to stimulate chondrocyte adhesion to collagen in the presence of metabolic inhibitors, while an activating anti-beta1 antibody was still effective. Thus, echistatin may promote cell adhesion by interfering with energy-dependent signals that keep the collagen receptor(s) in a low affinity state. Adhesion experiments performed in the presence of pharmacological inhibitors indicate that phosphatidyl inositol 3-kinase (PI3-K)/protein kinase C (PKC) and protein kinase A (PKA) pathways may transmit opposing signals on chondrocyte adhesion, and that collagen receptors are kept in a low-affinity state by PI3-kinase/PKC signalling. Since echistatin is a high-affinity ligand for alphavbeta3 integrin, the effect of the function-blocking anti-alphavbeta3 antibody LM609 was investigated. Like echistatin, LM609 stimulated chondrocyte adhesion to collagen and failed to support their attachment. Therefore, our data suggest that alphavbeta3-antagonists might regulate the binding activity of the beta1 collagen receptor, which in turn leads to the rapid transition of chondrocytes from suspension to an adherent state. PMID- 16270753 TI - Report of an independent peer review of an acrylonitrile risk assessment. AB - A peer review panel made up of experts in toxicology, epidemiology, cancer mode of action (MOA), cancer mechanisms, carcinogenicity, genotoxicity, dose-response, US Environmental Protection Agency (EPA) cancer and noncancer methods, pharmacokinetic modeling and acrylonitrile, met on 22-23 September 2003 in Cincinnati, OH. The purpose of the meeting was to provide an independent review of a risk assessment of acrylonitrile that had been prepared by the Acrylonitrile Group (AN Group). Toxicology Excellence for Risk Assessment (TERA) organized the peer review and selected the panel. The panel discussed the toxicity and epidemiology literature of acrylonitrile and MOA information, and reached conclusions regarding its MOA, weight of evidence (WOE) for carcinogenicity, preferred approach for dose-response assessment and risk values. This paper summarizes the discussion and conclusions of the panel regarding the acrylonitrile assessment. Subsequent to the peer review, the authors of the acrylonitrile assessment revised their report and the panel reviewed the revised report. A manuscript of the revised assessment is being published in Regulatory Toxicology and Pharmacology. PMID- 16270752 TI - Inhibitory effect of 14-3-3 proteins on serum-induced proliferation of cardiac fibroblasts. AB - Proliferation in cardiac fibroblasts (CFs) can be induced by a wide variety of growth factors that recruit multiple signal transduction pathways, including mitogen-activated protein kinase, phosphatidylinositol 3-kinase and protein kinase C. As a family of dimeric phophoserine-binding proteins, 14-3-3s are associated with a multitude of proteins that regulate signal transduction, apoptosis and checkpoint control pathways. However, it remains unknown whether the 14-3-3 proteins play an active role in cardiac proliferation and alter their expression patterns in response to growth factors in CFs. R18 peptide, an isoform independent 14-3-3 inhibitor, was used to disrupt 14-3-3 function by adenovirus mediated transfer of R18-EYFP (AdR18). Our results demonstrate that the 14-3-3 isoforms gamma, zeta and epsilon were highly expressed in CFs and the expression of 14-3-3 epsilon was elevated following serum stimulation. Inhibition of 14-3-3 proteins by AdR18 potentiated mitogen-induced DNA synthesis in CFs. This potentiation was presumably due to the increased inactivated glycogen synthase kinase-3 beta by Ser9 phosphorylation and nuclear factor of activated T-cell nuclear accumulation. However, AdR18 had no effect on extracellular signal regulated kinase phosphorylation and reduced p70 S6 kinase (p70S6K) phosphorylation upon mitogenic stimulation. Furthermore, though R18 can block 14 3-3 binding abilities, it did not affect the serum-induced upregulation of 14-3-3 epsilon protein. Collectively, these findings reveal that the expression of 14-3 3 epsilon can be upregulated by serum in CFs and 14-3-3s may exert an inhibitory effect on serum-induced proliferation. PMID- 16270754 TI - Prevention of acute cadmium toxicity by Picroliv. AB - The potential of Picroliv, a herbal extract against acute cadmium (Cd) intoxication, was evaluated in male rats. Biochemical and histopathological profile in rats pretreated with Picroliv (12 mg/kg, oral) followed by a single dose of Cd as cadmium chloride (CdCl2) (3 mg/kg, ip) revealed marked suppression of oxidative stress in liver and testes. The Cd-induced enhanced levels of lipid peroxidation, membrane fluidity and reduced levels of nonprotein sulphydryls and Na(+)K(+)ATPase were significantly restored to near normal by Picroliv pretreatment. In addition, the Cd-induced serum levels of glutamate oxaloacetate transaminase, glutamate pyruvate transaminase, gamma glutamyl transpeptidase and lactate dehydrogenase were restored to near basal levels. Hepatic and testicular histopathological damage was also minimized. The results strongly suggest definite hepato- and testicular protection by Picroliv. The antioxidant potential of the herbal extract in the major part, and not its chelating property, seems to be responsible for its ameliorative action. PMID- 16270755 TI - Classical radiation biology, the bystander effect and paradigms: a reply. AB - Although, in retrospect, it can be seen that the bystander effect and the related effect of genomic instability were observed well before they were recognized as such, they have not been able to be accommodated within the existing understanding of how radiation causes late effects, which provides the basis for radiological protection standards. It is argued here that before these effects can be fully researched and there can be full confidence in radiological protection, a paradigm shift that provides a framework in which these effects can be considered alongside the well established effects of radiation is needed. In particular this framework will encompass the epigenetic as well as genetic aspects of radiation biology. Examples of how this might be achieved are given. PMID- 16270756 TI - Colocalization of acetylcholinesterase, butyrylcholinesterase and choline acetyltransferase in rat spinal cord. AB - In our previous in situ hybridization study in the adult rat spinal cord we demonstrated that only neurons express mRNAs for acetylcholinesterase (AChE; EC 3.1.1.7.) and butyrylcholinesterase (BuChE; EC 3.1.1.8.) in this CNS region; no staining was detected for these two messages in the glia cells. Here we used immunocytochemical staining for choline acetyltransferase (ChAT; EC 2.3.1.6.) to answer the, as yet only briefly approached, question whether the cells expressing the messages for both cholinesterases are cholinergic. All neurons expressing mRNAs for AChE and BuChE were found to be ChAT positive. This finding supports the previous proposal that BuChE modulates cholinergic transmission in AChE knockout mice. PMID- 16270757 TI - The need for evidence in hypertension management: historical perspective. AB - The Task Force of the National Heart, Lung and Blood institute issued the first standardized, algorithmic approach to treating hypertension in 1973. The concept of a stepped-care approach was born at that time. Their initial recommendation for antihypertensive drug therapy was diuretics. Subsequent Joint National Committee (JNC) Reports on Detection, Evaluation, and Treatment of High Blood Pressure recommended that initial drug therapy be either a diuretic or beta adrenergic blocker, and then either of these two drugs, and then a calcium channel blocker (CCB) or an angiotensin-converting enzyme inhibitor (ACE inhibitors). The JNC-V then recommended any of the four classes or an alpha-beta blocker as initial therapy, but diuretics and b-blockers were preferable. That diuretics or beta-blockers should be the initial drug for noncomplicated hypertensive patient was also the recommendation of the Sixth Joint National committee report. Safety issues that arose after introduction ACE inhibitors and CCBs have since been mostly resolved. Drug treatment thresholds varied among the US, Canadian, British and WHO/ISH recommendations despite the fact that all were based on the same set of data. The concept of "the lower the blood pressure the better without causing symptoms" was the rule until the J-curve hypothesis emerged and generated a long debate. Now the current evidence supports the old concept, at least for some conditions such as hypertension in diabetic patients or in those with nephrotic-range proteinuria. Despite the repeated recommendations that thiazide-diuretics are preferred as the initial agent in hypertension treatment, many clinicians ignore these guidelines. This practice has added a signficant cost to hypertension treatment worldwide. PMID- 16270758 TI - Cyclosporine monotherapy for severe aplastic anemia: a developing country experience. AB - BACKGROUND: Immunosuppression is the most effective treatment for aplastic anemia after hematopoietic stem cell transplantation. Although the combination of cyclosporine and antithymocyte globulin (ATG) is superior to either agent alone, cyclosporine monotherapy is an easily available, safe and cheap immunosuppressive therapy (IST) option. These advantages are particularly valuable in developing countries where ATG is frequently not available. PATIENTS AND METHODS: In the referral hematology center in Yemen, 20 patients (16 males and 4 females) with severe aplastic anemia (SAA) were prospectively identified and managed with cyclosporine monotherapy during the period between April 2001 and November 2004. RESULTS: Data from 14 patients who received cyclosporine for at least 3 months were analyzed. At 6 months, 2 (14.3%) patients achieved complete remission (CR) and 5 (35.7%) patients achieved partial remission (PR) and at 1 year, 4 (28.6%) patients achieved CR and 3 (21.4%) patients remained in PR. The overall response rate was 50% and the cumulative survival rate at 1 year was 78.6%. The median time to remission was 120 days (range, 46 to 131 days). Side effects were modest and easily monitored. CONCLUSION: Our results support findings that cyclosporine monotherapy is an effective and safe immunosuppressive therapy for SAA, and that it could be a reasonable IST option for patients in developing countries. PMID- 16270759 TI - Mycophenolate mofetil in children with steroid-dependent and/or frequently relapsing nephrotic syndrome. AB - BACKGROUND: Mycophenolate mofetil (MMF) has emerged as a new agent for treatment of a variety of glomerular diseases. This study examines the safety and efficacy of MMF in treating pediatric patients with steroid-dependent (SD) and/or frequently relapsing (FR) nephrotic syndrome (NS). METHODS: We retrospectively reviewed the medical records of 18 patients with SDNS and/or FRNS treated with MMF for at least 3 months. MMF was used in 11 patients with SDNS (n=10) and FRNS (n=1), including 7 males and 4 females. RESULTS: Mean age at time of diagnosis of NS was 3.3 years (range, 1.1-8.5 years), and at the start of MMF 5.9 years (range, 2.9-10 years). Seven patients had a renal biopsy prior to starting MMF; all had mesangial proliferative glomerulonephritis. Mean follow-up after starting MMF was 12.2 months (range, 4-24 months). Mean MMF dose was 948 mg/m2/day (range, 500-1087 mg/m2/day). MMF resulted in improvement in 9 of 11 patients, with 8 patients weaned off steroids completely, with a reduction in the mean relapse rate from 4.7 relapses/patient/year (range, 2.4-6) before MMF to 1.05 relapses/patient/year (range, 0-4.5) after MMF therapy (P=0.0001). The relative risk for relapse before MMF was 4.7 (P=0.0002). None of the patients had significant adverse events or intolerance to MMFtherapy. CONCLUSION: We conclude that MMF is a safe and effective option for treatment of children with SDNS and/or FRNS. PMID- 16270760 TI - Cleft lip and palate in southwestern Iran: an epidemiologic study of live births. AB - BACKGROUND: Cleft lip with or without cleft palate (CL/P), is the most common congenital anomaly in the head and neck worldwide. We studied the prevalence of cleft lip (CL) and palate (CP) in Shiraz, which is located in southwestern Iran. METHODS: Data was collected from the teaching hospitals of the Shiraz University of Medical Sciences. Records of 147,608 consecutive live births delivered in three hospitals during the 10 years from November 1993 to November 2003 were reviewed. RESULTS: One hundred nineteen cases of CL/P or CP alone were registered for a prevalence of 0.80 per thousand live births. The overall male/female ratio was 1.25. The male/female ratio was 1.22 in the CL patients and 1.58 in the CL/P patients. A male predominance was obvious in both groups. CONCLUSION: This study shows that the prevalence rate of CL and CP in southwestern Iran is closer to the low prevalence reported in African countries and is lower than the prevalence reported in previous reports in Caucasians. PMID- 16270761 TI - Childhood obesity in Iraq: prevalence and possible risk factors. AB - BACKGROUND: Childhood obesity is increasing in prevalence in developing countries. We conducted this study to assess the prevalence of overweight and obesity in primary school children age 7 to 13 years in central Iraq and to identify possible risk factors. METHODS: The study was conducted in Babil governorate from October through March 2002. Eight thousand three hundred pupils (4100 girls and 4200 boys) randomly chosen primary schools were included in the study. BMI was measured and compared to standard tables. Information on possible risk factors was collected by questionnaire. RESULTS: The prevalence of overweight and obesity was 6% and 1.3%, respectively. Significant associations were found between overweight and age, residency (urban-rural), birth rank, breast-feeding and certain dietary and lifestyle practices. CONCLUSION: The prevalence of obesity and overweight is relatively high in children in central Iraq. Modern dietary habits are an important influence in the development of childhood obesity that should be watched carefully and controlled. PMID- 16270762 TI - Prevalence of differentiated thyroid cancer in 810 cases of surgically treated goiter in Yemen. AB - BACKGROUND: In 1990, the World Health Organization (WHO) suggested that severe iodine deficiency exists in Yemen. Therefore, we looked at the prevalence of differentiated thyroid goiter in 810 cases consecutively treated by surgery for goiter. METHODS: This was a retrospective study of 810 surgically operated cases of goiter over a 5-year period (1999-2003). All cases were evaluated on H&E stained sections from embedded, 10% buffered formalin fixed tissue blocks. Special stains and immunohistochemical analysis were done in Yemen and abroad. Most patients were older than 20 years of age and were from the high altitude areas (2000 to 2600 meters above sea level), where iodine deficiency disorders (IDD) are well documented. RESULTS: In the 810 cases, 729 (90%) were females and the remaining 81 (10%) were males, with female-to-male ratio of 9:1. Differentiated thyroid cancer (DTC) was found in 170 (21%) cases, including 148 (86.4%) females and 22 (13.6%) males. Nearly 60% of the cases were in the age group of 21-40 years. Papillary carcinoma was the most common type of DTC (164 cases, 96.5%). CONCLUSIONS: In a Yemeni population, which has a high prevalence of iodine deficiency, 21% of patients operated on for nodular goiter without pre operative fine needle aspiration biopsy had thyroid cancer, mostly of the papillary type. In this study, males and elderly patients with goiter had a higher chance of having malignancy. PMID- 16270763 TI - Malignant lymphoma in Jordan: a retrospective analysis of 347 cases according to the World Health Organization classification. AB - BACKGROUND: Most studies describing the subtypes of lymphoma in Jordan were carried out in the 1980s at a time when immunohistochemical facilities were unavailable. Using a database established after immunohistochemical studies were introduced, we determined the frequency of the various types of nodal and extranodal lymphomas in the adult and paediatric Jordanian population. We also assessed the incidence of bone marrow involvement at the initial presentation for each lymphoma type. METHODS: A retrospective analysis of the histopathological subtypes of various lymphomas was conducted on all primary lymphoma cases diagnosed during a 3-year period between January 2001 and December 2003. RESULTS: Of 347 patients included in the study, 78.4% had non-Hodgkin's lymphoma (NHL) and 21.6% had Hodgkin's lymphoma (HL). In the NHL group, diffuse large B-cell lymphoma was the most common (28.2%) followed by follicular lymphoma (15.6%). In the HL group, the nodular sclerosis variant was the most frequent (7.8%) followed by the mixed cellularity type (5.5%). Of all the lymphoma cases, the highest incidence of marrow involvement was seen in patients with lymphoplasmacytic lymphoma. Forty-nine patients were children (age <15 years) in whom Burkitt's lymphoma (15 cases) and HL (14 cases) were the commonest subtypes. One-hundred six patients with primary extranodal lymphomas (ENL) accounted for 30.5% of all lymphomas. CONCLUSIONS: There is a higher incidence of NHL in Jordan compared with other series in the Middle East. Among the various lymphomas, diffuse large B-cell lymphoma is the most commonly encountered lymphoma in adults. Burkitt's lymphoma and Hodgkin's disease are the most frequent childhood lymphomas, followed closely by lymphoblastic lymphoma. PMID- 16270764 TI - A descriptive study of medical malpractice cases in Turkey. AB - BACKGROUND: Medical malpractice claims in Turkey have increased. We evaluated the problem by describing medical malpractice cases assessed by the Higher Health Council between 1993 and 1998. Our recommendations should help to improve care and decrease medical malpractice claims. METHODS: We reviewed 997 medical malpractice cases reported to the Higher Health Council between 1993 and 1998 and examined the decisions made by the Higher Health Council. We collected data on demographic characteristics, such as the type of the institution where the defendants worked, type of medical malpractice, and medical outcome. RESULTS: There were 997 medical malpractice cases reported to the Higher Health Council in the six years between 1993 and 1998. The Higher Health Council decided that 47.7% of the physicians were liable. Malpractice cases were mostly seen in state hospitals (42.4%). Fifty-nine percent of the cases resulted in death. Among actions that led to malpractice lawsuits against all health care workers, including physicians, the most common were negligence, inappropriate treatment, and diagnostic failure. CONCLUSION: We think it is necessary to revise the health system and working conditions in hospitals and to develop clinical practice guidelines. We are of the opinion that an emphasis on the use of diagnosis and therapy protocols, standards, post-graduation education, clear and informed patient consent, and improved communication with patients will drastically decrease medical malpractice claims. PMID- 16270765 TI - In vitro activity of ciprofloxacin, ofloxacin and levofloxacin against Mycobacterium tuberculosis. AB - BACKGROUND: The increasing incidence of drug-resistant Mycobacterium tuberculosis necessitates therapeutic alternatives. The fluoroquinolones fulfill most of the criteria for an ideal class of antimycobacterial drugs. The aim of the present study was to determine to in vitro activities of ciprofloxacin, ofloxacin, and levofloxacin against M. tuberculosis strains. METHODS: Susceptibility to four antituberculous drugs used in first-line treatment of tuberculosis was tested in 100 strains isolated from clinical samples. Nineteen strains (19%) were resistant to at least one of the four antituberculous drugs and 13 were multidrug resistant. The in vitro antimycobacterial activity of ciprofloxacin, ofloxacin, and levofloxacin was then determined against 100 M. tuberculosis strains using standard agar proportion dilution method. RESULTS: Ciprofloxacin, ofloxacin, and levofloxacin were active against all tested strains of M. tuberculosis in vitro. CONCLUSIONS: Ciprofloxacin, ofloxacin, and levofloxacin have relatively potent in vitro activity against M. tuberculosis. Further in vivo studies are needed to determine the role of these compounds in the treatment of tuberculosis, but use should be limited to special circumstances rather than first-line treatment. PMID- 16270766 TI - Ruptured extrauterine gestation in heterotopic pregnancy. PMID- 16270767 TI - Calciphylaxis in a 33-year-old man with end-stage renal disease. PMID- 16270768 TI - Shigella flexneri perinephric abscess and bacteremia. PMID- 16270769 TI - Plantar keratoderma: a manifestation of tyrosinemia type II (Richner-Hanhart syndrome). PMID- 16270770 TI - Helicobacter pylori eradication for the treatment of dyspeptic symptoms in chronic renal failure. PMID- 16270771 TI - Unusual presentations of differentiated thyroid cancer: analysis of 55 cases from North India. PMID- 16270772 TI - Periodic fever, rash and arthropathy in a child. PMID- 16270773 TI - Surgical pearl: A disposable syringe barrel is a better alternative to a Foley's catheter for intraoperative nasal ala stabilization. PMID- 16270774 TI - Value of ophthalmic features as a means of diagnosis of HIV/AIDS infection. PMID- 16270775 TI - Large cell calcifying Sertoli cell tumor of the testis. PMID- 16270776 TI - Left mesocolic hernia presenting as an abdominal lump in an adult. PMID- 16270777 TI - Re: Nutritional koilonychia in 32 Iraqi subjects. PMID- 16270778 TI - The International Survey on health professionals and tobacco shows 32% of health professionals smoke, 10% quit. PMID- 16270779 TI - Carotid angioplasty for the management of extracranial carotid occlusive disease. AB - INTRODUCTION: Carotid endarterectomy (CEA) has become established as the preferred approach to the management of critical carotid stenosis. Carotid angioplasty with stenting (CAS) has recently arisen as an alternative in the treatment of carotid occlusive disease. This report describes our experience with carotid angioplasty applied to an unselected patient population suffering from high-grade carotid occlusive disease. METHODS: All patients suffering from carotid stenosis (> 50% symptomatic or > 80% asymptomatic) were offered CAS or CEA. The first 39 patients who underwent attempted CAS over this last year are reported here. CAS was performed with the SMART PRECISE or ACCULINK stents. All procedures were performed with cerebral protection. RESULTS: The planned procedure success rate was 97% and the major adverse event (MAE) rate was 2.6% in 38 patients who underwent successful CAS. This included a minor stroke and a subendocardial myocardial infarction in the same individual. Both events were attributed to sustained postprocedural hypotension probably induced by increased carotid sinus activity. CONCLUSION: CAS can be accomplished with a MAE rate comparable to CEA and will likely become the dominant alternative to CEA for the management of carotid stenosis. In the setting of equivalent morbidity, it appears likely that a nonsurgical option will be preferred by patients. PMID- 16270780 TI - Intramural hematoma of the descending thoracic aorta. AB - Aortic intramural hematoma (AIH) is part of the acute aortic syndrome together with aortic dissection and penetrating aortic ulcer. AIH represents up to 12% of acute aortic syndrome cases and has a clinical presentation indistinguishable from that of classic aortic dissection. A patient with AIH is presented and the diagnostic, prognostic and therapeutic implications of the disease are discussed. PMID- 16270781 TI - Synchronous lung cancers: case report and review of the literature. AB - Lung cancers may present separately as synchronous lesions and can pose a diagnostic and therapeutic dilemma. A 75-year-old man is reported who presented with two synchronous lung cancers, in his left upper and lower lobes, respectively. Extensive radiologic evaluation and mediastinoscopy failed to detect any evidence of metastatic disease. He underwent lung preserving surgery consisting of wedge resection and segmentectomy. Unfortunately, this patient relapsed six months after surgery. Long-term survival after surgical resection has been reported in such instances indicating the need for an individualized approach to treatment. PMID- 16270782 TI - Endovascular repair of transplant nephrectomy external iliac artery pseudoaneurysm. AB - The formation of a pseudoaneurysm at the site of transplant nephrectomy is a rare complication, having been reported only six previous times in the literature. Possible causes of this occurrence include infection or rejection of the anastomotic remnant. Diagnosis of the pseudoaneurysm with radiological imaging is necessary. The first successful repair of a noninfected pseudoaneurysm of a transplant nephrectomy patient is described using an endovascular graft. PMID- 16270784 TI - How dangerous are thyroid nodules? 1955. PMID- 16270783 TI - The Medical Examining Board speaks on pain management: everyone should listen. AB - PURPOSE: To alert physicians to the Statement of the Connecticut Medical Examining Board on the Use of Controlled Substances for the Treatment of Pain and provide a historical account of events leading to the promulgation of the Statement. FINDINGS: Studies conducted over the past five years have identified issues on pain control requiring attention. Established medical organizations and regulatory boards in Connecticut are attempting to address patient needs for treatment of pain while clarifying expectations of physicians, thereby reducing the likelihood of inappropriate pain treatment and licensing sanctions. CONCLUSION: All physicians should read and become familiar with the Statement of the Connecticut Medical Examining Board on the Use of Controlled Substances for the Treatment of Pain. PMID- 16270785 TI - Quality, safety, and the language of adverse events. PMID- 16270786 TI - Say goodbye to the low profile. PMID- 16270787 TI - Golden excellence. PMID- 16270789 TI - Targeting symptom domains: a strategy for pharmacotherapy in childhood pervasive developmental disorders. AB - This article summarizes clinical guidelines and reviews psychopharmacologic studies involving children and adolescents with Pervasive Developmental Disorders (PDDs). Strategies are drawn from basic principles of pediatric psychopharmacology, nonpharmacological approaches to the treatment of PDDs, and practical clinical experience in an attempt to provide the practitioner with an evidence-based approach to utilizing pharmacotherapy in children and adolescents with PDDs. Although early identification followed by intensive educational and behavioral interventions remains the essential treatment of PDDs, there is evidence to support the identification of target behaviors or symptom domains, for medication. Evidence-based medicine (EBM) is then used to guide pharmacotherapy with the overall goal of optimizing the benefits of multidisciplinary treatment interventions. PMID- 16270788 TI - Motivational interviewing techniques and the harm-reduction model in a short-term substance-abuse group for adolescents with psychiatric problems. AB - The authors describe techniques and practical examples for working with substance abusing teens within a short-term psychiatric setting that derive from the Harm Reduction model and the Motivational Interviewing approach. PMID- 16270790 TI - Schools and mental health: an evolving partnership. AB - The US Surgeon General's Report (1999) studied the prevalence of mental illness in school-aged children and adolescents. It concluded that the national prevalence rate of students with treatable mental health problems was 20%-25%. The report noted that an astounding 90% of children go undetected or undertreated in terms of their mental health needs. Schools, by their very nature, are ideal settings for primary prevention. They are structured to allow for professional observation of students' behaviors, over time, within a caring learning environment. School staff can be trained to observe signs and symptoms and refer them for professional diagnostic and treatment services when indicated. Three specialized programs demonstrate how timely observations and referrals for treatment by school professionals improve students' access to psychiatric care. PMID- 16270791 TI - Risk of suicide and SSRIs. AB - The controversy surrounding the use of Selective Serotonin Reuptake Inhibitors (SSRIs), in the treatment of depression and other psychiatric disorders in children and adolescents, has come to the forefront since the question of increased suicidal risk has been raised. There are limited research data available to thoroughly analyze the safety and efficacy of this class of medications in youths. In this article, we review the evidence available, the events that led to the black box warning, and the practical implications of the warning for providers, youth and families. PMID- 16270792 TI - Child and adolescent psychiatry training in the 21st century. AB - The mental health needs of the nation's children, adolescents and families are not being met for a variety of social, economic, cultural and political reasons. A shortage of children's mental health providers, including child and adolescent psychiatrists, is one aspect of the problem. Barriers to recruitment into the field of Child and Adolescent Psychiatry (CAP) and proposed initiatives at the local and national levels to help alleviate the work force shortage are discussed. A description of the child and adolescent psychiatrist and the training requirements are provided. A challenge to training programs is to prepare graduates who are competent to provide critical mental health care to our youth and their families in the 21st century, while preserving the unique qualities of our profession. PMID- 16270794 TI - Functional neuroimaging in child and adolescent psychiatry. PMID- 16270793 TI - An overview of evidence-based psychotherapy for children and adolescents. AB - Research on psychotherapy outcomes, particularly with children, has a short but evolving history. Studies in the 1950s and 1960s suggested therapy was no more useful than no treatment and the passage of time. There were many flaws in the research upon which this conclusion of no improvement from psychotherapy was based and it has taken the next half century to generate more studies and to reconsider the evidence for efficacy of psychotherapy. While problems continue to exist with the quantity, strength, and generalizability of research on child psychotherapies, it is increasingly accepted that efficacious treatments do exist for child and adolescent disorders. This article provides an overview of evidence based psychotherapies for the two chief domains of psychiatric problems in children and adolescents: externalizing and internalizing disorders. The concurrent challenge of integrating research findings into typical treatment settings and promoting their adoption with clinical practitioners will be highlighted. PMID- 16270795 TI - A golden anniversary: fifty years of child and adolescent psychiatry at the Institute of Living. PMID- 16270796 TI - Nursing care of the psychiatric patient in the general hospital. 1955. PMID- 16270797 TI - A word on death certificates. PMID- 16270798 TI - A boy named Luke. PMID- 16270799 TI - Influence of latent 'asymptomatic' toxoplasmosis on body weight of pregnant women. AB - The latent toxoplasmosis is usually considered to be asymptomatic, however, this paradigm has never been rigorously tested. Here we searched for symptoms of deterioration of physical health (decrease of weight) in infected people by analysis of clinical records of 758 women tested for toxoplasmosis in the 16th week of gravidity. Toxoplasma-positive women have a lower body weight in the 16th week of gravidity (p = 0.02) than Toxoplasma-negative women. Moreover, a negative correlation between weight and the duration of toxoplasmosis was found in a subset of 174 Toxoplasma-positive women (p = 0.04), suggesting that slow and cumulative effects of latent toxoplasmosis, rather than a transient effect of acute toxoplasmosis, are responsible for the decreased weight of infected subjects. Longer duration of gravidity estimated from the date of last menstruation in the set of Toxoplasma-positive women in the 16th week of gravidity estimated with ultrasonography (p = 0.04) suggests a possibility of retarded foetal growth in Toxoplasma-positive women. The prevalence of latent toxoplasmosis is extremely high. Therefore, even its mild symptoms such as the decreased body weight in Toxoplasma-positive pregnant women might in fact indicate an unrecognized serious public health problem. PMID- 16270800 TI - Mice serve as paratenic hosts for the transmission of Caryospora duszynskii (Apicomplexa: Eimeriidae) between snakes of the genus Elaphe. AB - Caryospora duszynskii Upton, Current et Barnard, 1984 was successfully transmitted to snakes of the genus Elaphe by feeding them previously infected mice. Fifty thousand oocysts were orally administered to two mouse strains, BALB/c and Crl:CD-1(ICR)BR, which were subsequently fed to captive-born coccidia free Elaphe guttata (L.) in two respective independent experiments. Both E. guttata expelled C. duszynskii oocysts in their faeces, beginning on day 18 and 26 post infection (p.i.) and shed oocysts continuously through the end of the experiment, day 230 and 135 p.i., respectively. There were no parasitic stages or lesions in mice, as revealed by histological examination. Experiments proved that rodents serve as paratenic hosts for C. duszynskii. In summary we discuss the life-cycle strategies of Caryospora spp. in reptiles and present three general modes of their development. PMID- 16270801 TI - Morphology and aspects of growth of a trypanosome transmitted by the marine leech Johanssonia arctica (Piscicolidae) from Northern Norway. AB - The fish leech Johanssonia arctica (Johansson, 1898) was collected from king crabs Paralithodes camtschaticus (Tilesius, 1815) in Finnmark, N Norway, and allowed to feed on experimental fish hosts in the laboratory. This leech ingested blood from laboratory-reared cod (Gadus morhua) and halibut (Hippoglossus hippoglossus). Some experimental halibut acquired trypanosome infection, with parasitaemia between ca. 500 and 60,000 trypanosomes ml(-1). The trypanosomes were of variable size and measured 39-90 microm (mean 57 microm) ca. 81 days post infection. Characteristic features are cell striation, refractile cytoplasmic granules, anterior nucleus and a relatively long (ca. 6 microm, max 9 microm) distance from the posterior end to the kinetoplast. Following growth, the trypanosomes became increasingly slender, with fewer striae and a nucleus position less pronounced anterior. The trypanosome is considered distinct from a type transmitted by the leech Calliobdella nodulifera (Malm, 1863) in the NE Atlantic, but is regarded conspecific with a trypanosome transmitted by J. arctica in the NW Atlantic. This trypanosome has in the past been identified as Trypanosoma murmanensis Nikitin, 1927, a poorly described species. T. murmanensis cannot be recognized with certainty among the trypanosomes transmitted by C. nodulifera and J. arctica respectively. We propose that the J. arctica transmitted species is considered T. murmanensis Nikitin, 1927 sensu stricto. PMID- 16270802 TI - Supplemental diagnosis and molecular taxonomy of Myxobolus diaphanus (Fantham, Porter et Richardson, 1940) (Myxozoa) parasitizing Fundulus diaphanus (Cyprinodontiformes) in Nova Scotia, Canada. AB - Myxobolus diaphanus (Fantham, Porter et Richardson, 1940) was found in banded killifish Fundulus diaphanus (Lesueur) at several freshwater localities in Nova Scotia, including the type locality at the mouth of the Salmon River, Guysborough County. The new material, the first to be reported in 64 years, was used to supplement information on spore morphology, to document the site of development in the tissue, and to compare sequence data of the 18S rDNA to other studied myxobolids. Plasmodia with developed spores occurred in loose connective tissue of the head, the dermis (particularly in the roof of the mouth and at the base of fins), surface of the brain and ovary, muscle epimysium, and the submucosa of the intestine. Developed plasmodia containing spores were also found free in the lumen of the vena cava and within fluid-filled spaces of the skull, mandible and lower jaw. A phylogenetic analysis using 18S rDNA (878 bp) placed M. diaphanus in a terminal clade containing certain freshwater species of Henneguya, all of which occur in North America and have elongate spore bodies. PMID- 16270803 TI - Branchotenthes octohamatus sp. n. (Monogenea: Hexabothriidae) from the gills of the southern fiddler ray, Trygonorrhina fasciata (Rhinobatidae) in South Australia: description of adult and larva. AB - Branchotenthes octohamatus sp. n. (Monogenea: Hexabothriidae) is described from the gills of the southern fiddler ray, Trygonorrhina fasciata Muller et Henle (Elasmobranchii: Rhinobatidae), off Adelaide, South Australia. It is distinguished from the type species, Branchotenthes robinoverstreeti Bullard et Dippenaar, 2003, by producing eggs that are joined end to end forming a chain, in the morphology of the male copulatory organ that has a pronounced constriction in duct diameter between proximal and distal regions, the possession of a thin muscular layer surrounding the proximal part of the male copulatory organ and distal region of the vaginae, and by the absence of a raised process on the shaft of the hamulus. An amended generic diagnosis is provided and the reliability of sperm duct number as a generic character is discussed. The oncomiracidium of B. octohamatus is also described and is the first monogenean to be described with only eight hooklets in the larval haptor. This discovery of eight hooklets may be important for higher-level monogenean evolutionary hypotheses. PMID- 16270804 TI - Description of Pseudorhabdosynochus seabassi sp. n. (Monogenea: Diplectanidae) from Lates calcarifer and revision of the phylogenetic position of Diplectanum grouperi (Monogenea: Diplectanidae) based on rDNA sequence data. AB - Pseudorhabdosynochus seabassi sp. n. (Monogenea: Diplectanidae) from the gill filaments of Lates calcarifer Bloch, a marine teleost fish held in floating sea cages in Guangdong Province, China, is described based on morphological observations and molecular data. The shapes of the male copulatory organs (MCO) of Pseudorhabdosynochus spp. were the focus of this study. The typical proximal part of the MCO in most species of Pseudorhabdosynochus is reniform, heavily sclerotized, and divided into four chambers. However, the new species from L. calcarifer has a bulbous proximal region with four concentric layers of apparent muscular origin, instead of a reniform structure with four compartments. This organ is also different in Diplectanum grouperi Bu, Leong, Wong, Woo et Foo, 1999, being sclerotized, cup-shaped, wide proximally with four concentric muscular layers and tubular distally. The 3' terminal portion of the small subunit ribosomal RNA gene (ssrDNA) and the 5' terminal region (domains C1-D2) of the large subunit ribosomal RNA gene (lsrDNA) were used to reconstruct the phylogenetic relationships of P. seabassi and D. grouperi with related taxa utilizing maximum-parsimony and neighbour-joining methods. Phylogenetic analyses unequivocally placed D. grouperi amongst Pseudorhabdosynochus using either ssrDNA or lsrDNA data. All species of Pseudorhabdosynochus (including D. grouperi) used in this study clustered together, inferring monophyly. Based on molecular phylogenetic evidence, we propose that D. grouperi from Epinephelus coioides Hamilton be transferred to Pseudorhabdosynochus as P. grouperi comb. n. PMID- 16270805 TI - Fine structure of the male reproductive ducts, vagina and seminal receptacle of Cyathocephalus truncatus (Cestoda: Spathebothriidea). AB - Fine structure of the vas efferens, vas deferens, ejaculatory duct with accessory glands and vagina with seminal receptacle is described in the spathebothriidean tapeworm, Cyathocephalus truncatus (Pallas, 1781) Kessler, 1868. The numerous well-developed prostate glands are characterised by having secretory granules with an electron-dense core surrounded by a matrix of lower electron density. Coalescence of the outer part of the granules with each other takes place in the terminal end of the secretory ducts. The position of prostate glands around the proximal part of the cirrus pouch and terminating in the ejaculatory duct is a characteristic feature of the Spathebothriidea. Up to 20 closely arranged muscle layers make up the muscular cirrus pouch wall with 4 well-developed muscular layers in the ejaculatory duct and cirrus. Both the cirrus and the vagina are covered with the same uniform cone-shaped microtriches. The vagina has an extensive seminal receptacle. All of these structures are well-adapted to insure successful sperm transfer involving ejaculation and storage, probably for both self- and cross-insemination. Cyathocephalus truncatus has a cirrus similar to that of the monozoic, progenetic caryophyllidean, Archigetes sieboldi and well developed prostate glands like those of the polyzoic pseudophyllidean, Diphyllobothrium latum. The ultrastructural aspects of the male and female reproductive system of C. truncatus are compared with those of other tapeworms. PMID- 16270807 TI - Data on the morphology of Viguiera dicrurusi Gupta, 1960 (Nematoda: Habronematidae) with notes on related forms. AB - Morphology of the nematode Viguiera dicrurusi Gupta, 1960 harboured by Dicrurus macrocercus albirictus (Hodgson) (Passseriformes: Dicruridae) from Baruipara in 24-Pargonas (South) district, West Bengal, India was studied by light and scanning electron microscopy (SEM). This represents the first study of V. dicrurusi using SEM. Scanning electron micrographs provided detailed information about the nature of pseudolabial plates, number and shape of teeth, dentate nature of striae, and the relative position of vulva, anus and phasmid opening in female. A detailed morphometrical comparison of this species with Viguiera viduae Chabaud, 1960 described from Dicrurus forficatus from Madagascar indicates that V. viduae is a junior synonym of V. dicrurusi. Two other species, Viguiera bhujangai Jehan, 1972 and Viguiera adsimilisai Sood et Kalia, 1978 are considered species inquirendae. PMID- 16270806 TI - Redescription of Eubothrium fragile (Rudolphi, 1802) and E. rugosum (Batsch, 1786) (Cestoda: Pseudophyllidea), parasites of fish in the Holarctic Region. AB - Two fish cestodes, the little-known Eubothrium fragile (Rudolphi, 1802) and E. rugosum (Batsch, 1786), the type species of the genus Eubothrium Nybelin, 1922, are redescribed on the basis of new material from twaite shad, Alosa fallax (Lacepede, 1803), from England and burbot, Lota lota (Linnaeus, 1758), from Russia, respectively. The tapeworms are compared with two other species of the genus, E. crassum (Bloch, 1779) and E. salvelini (Schrank, 1790), common parasites of salmonid fish in the Holarctic. The most notable differential characters are the size and the shape of the scolex (smaller and oval in E. fragile), the shape of the apical disc (four or more indentations in E. crassum), the number and size of the testes (the largest and least numerous in E. rugosum), and the position and size of the vitelline follicles (almost entirely cortical in distribution in E. fragile and E. crassum versus largely medullary in E. rugosum and E. salvelini). A comparison of species has also shown the morphological similarity of the freshwater species (E. rugosum and E. salvelini) on one hand and those of marine origin, E. fragile and E. crassum, on the other, with the latter species occurring also in fresh waters. A key to the identification of the species studied is also provided. PMID- 16270808 TI - Two new species of Philometra (Nematoda: Philometridae) from needlefishes (Belonidae) in Iraq, with a key to Philometra spp. parasitic in the host's subcutaneous tissue, fins and musculature. AB - Two new nematode species, Philometra strongylurae sp. n. and Philometra tylosuri sp. n., are described from female specimens collected from needlefishes (Belonidae, Beloniformes) off the Fao coast, Basrah, Persian Gulf, in Iraq. Philometra strongylurae (gravid females with larvae) was recorded from the subcutaneous tissue, muscles of beak and gills of Strongylura leiura (type host) and S. strongylura, whereas P. tylosuri (nongravid females) from the musculature and the subcutaneous tissue of Tylosurus crocodilus. The former is characterized mainly by the presence of eight markedly large cephalic papillae, conspicuously large anterior oesophageal bulb and by the length of body (15-20 mm); the latter by eight minute cephalic papillae, poorly developed oesophageal bulb and by the body length (46-72 mm). A key to Philometra spp. with gravid females parasitic in the subcutaneous tissue, fins or musculature of fishes is provided. PMID- 16270809 TI - Babesia microti (Piroplasmida: Babesiidae) in nymphal Ixodes ricinus (Acari: Ixodidae) in the Czech Republic. AB - A total of 350 nymphs of the common tick Ixodes ricinus (Linnaeus, 1758) were collected in an endemic focus of Lyme borreliosis (South Moravia, Czech Republic) and examined for the presence of the protozoan Babesia microti (Franca, 1909) by polymerase chain reaction (PCR), using primers specific for the B. microti gene encoding small subunit rRNA. The assay revealed five positive pools (out of 70 pools examined); the corresponding prevalence rate was about 1.5%. Sequence analysis of the PCR products confirmed their 100% homology with that of B. microti. The study represents the first evidence of B. microti in ixodid ticks in the Czech Republic. PMID- 16270810 TI - Intraerythrocytic merogony in Haemogregarina koppiensis (Apicomplexa: Adeleorina: Haemogregarinidae). AB - During October 2003, a specimen of Amblyrhynchotes honckenii (Bloch, 1795) was captured at low tide, with a hand net, in a rock pool at Koppie Alleen, De Hoop Nature Reserve, South Africa. This fish was heavily parasitized by unidentified gnathiid praniza larvae, caligid copepods identified as Caligus tetrodontis Barnard, 1948, cymothoid isopods identified as Cinusa tetrodontis (Schioedte et Meinert, 1884), and the blood protozoan Haemogregarina koppiensis Smit et Davies, 2001. Giemsa-stained blood smears from this fish revealed new and unusual stages of merogony for H. koppiensis that included small, rounded, likely intraerythrocytic merozoites arranged in circles of eight around the host nucleus. Host cells appeared ghost-like and enlarged compared with normal erythrocytes. Identical merozoites, usually in clusters of up to 16, were also observed free of host cells. The pattern of merogony seen in H. koppiensis is unusual for a fish haemogregarine. PMID- 16270811 TI - A 100 year legacy of care. PMID- 16270813 TI - A tale of two practicums. A great beginning for a satisfying career. PMID- 16270812 TI - A collaborative approach to diabetes nephropathy prevention. AB - Diabetic nephropathy is one of the major complications of diabetes and is the leading cause of end-stage renal disease (ESRD) (40 percent). Diabetes and its complications take a huge toll on the quality of life of those affected. There are substantial costs for both individuals and the health-care system and projections of the increasing incidence of diabetes over the next 12 years serve to increase the sense of urgency of efforts to prevent diabetes and its complications. For people who develop diabetic nephropathy, the kidneys gradually lose the ability to function properly. Treatment of ESRD by renal replacement therapy (RRT), e.g. dialysis, results in a decreased quality of life and is costly and time consuming. However, clinical practice guidelines (CPGs) identify and provide evidence for early management and treatment to prevent diabetic nephropathy as well as heart attacks, strokes and retinopathy. PMID- 16270814 TI - A tale of two practicums. An early lesson on a presence experience. PMID- 16270815 TI - [The non-neuronal cholinergic system in immunocompetent cells]. AB - Most of the cholinergic components, which are expressed in neurons, exist in the immunocompetent cells, especially the lymphocytes, and thus constitute an independent cholinergic system. Comparing with neuronal cholinergic system, the non-neuronal cholinergic system has its unique characteristics, and may be involved in regulation of immune function and inflammatory responses in human and other mammals. An insight into the non-neuronal cholinergic system in immunocompetent cells may facilitate further understanding of pathophysiology and novel therapeutics of immune abnormalities, infectious diseases and neurodegenerative diseases. PMID- 16270816 TI - [DREAM/Calsenilin/KChIP3: a new multifunctional protein in nervous system]. AB - A newly found transcription factor-DREAM can bind specifically with downstream regulatory element of genes, such as PPD, Hrk and c-Fos, and repress gene expression. It represents the first known Ca2+ binding protein to function as a DNA-binding transcription regulator. This provides a new mechanism of Ca2+ dependent regulation of gene expression in addition to protein kinase/phosphatase signaling pathway. In fact, DREAM, calsenilin and KChIP3, which are identified by different laboratories, are the same protein. So DREAM is a multifunctional protein, including PS interacting protein, Kv4 regulatory protein and transcription factor. The distribution, function and regulation of DREAM, as well as relationship between DREAM and pain, are reviewed in the present article. PMID- 16270817 TI - [Cytological basis and significance of mesenchymal stem cells differentiated into endothelial cells]. AB - Mesenchymal stem cells (MSCs) are a multipotent population which mainly localized in the bone marrow. MSCs were also isolated from the umbilical blood, peripheral blood, fatty tissue, skin and so on. MSCs have special immunity and persist in the xenogeneic explantation which expand the clinical application. At present, the system of culturation, induction and identification of MSCs are gradually becoming mature. Meanwhile, MSCs differentiate into cells derived from mesoderm in the theory. Endothelial cells derived from the mesoderm, MSCs have potency of differentiated into endothelial cells (ECs). This review focuses on the significance and cytological basis of human MSCs differentiate into endothelial cells, as well as the recent advancement and application on the tissue engineering. PMID- 16270818 TI - [Regulation of prostaglandin E molecular network in mammalian reproduction]. AB - Prostaglandin E molecular network includes cyclooxygenase, phospholipase A2, prostaglandin E, prostaglandin E receptors, and prostaglandin E synthase. Recently, it was found that PGE2 receptors localized not only on the cell membrane but also on the envelope of nucleus. They are different in the signaling pathways and in regulation mechanism between nuclear receptors and membrane receptors. They compose a delicate network, which plays important roles in mammalian reproduction, especially in most processes of female reproduction. PMID- 16270819 TI - [The structure, classification and function of RGS proteins]. AB - Regulators of G protein signaling (RGS) are a family of highly diverse, multifunctional signaling proteins that bind directly to activated Galpha subunits and markedly stimulates the GTPase activity of Galpha subunits leading to their deactivation and termination of downstream signals. RGS family members share a conserved 130-amino acid core domain which is responsible for binding Galpha subunits and negatively regulate G protein signal. Many RGS proteins possess additional non RGS domains and motifs. The non RGS domains are involved to integrate different G protein signaling pathways and as a saffolding protein that links G protein to related signaling proteins. PMID- 16270820 TI - [Cytoglobin: the fourth kind of oxygen-binding globin]. PMID- 16270822 TI - [Remote preconditioning in the heart]. PMID- 16270821 TI - [Apelin and its biological effects]. PMID- 16270823 TI - [Diazepam-binding inhibitor]. PMID- 16270825 TI - [The two-edged sword to cancer--protein Rad51D]. PMID- 16270824 TI - [Iron metabolism and iron-regulatory hormone, hepcidin]. PMID- 16270826 TI - [The structure and biological functions of A-kinase anchor proteins]. PMID- 16270827 TI - [Progress in the gene polymorphisms of drug-related transporters]. PMID- 16270828 TI - [Wnt signaling pathway and neural stem cells]. PMID- 16270830 TI - [Leptin and wound healing]. PMID- 16270829 TI - [Advances in vulnerable mechanisms of selectively vulnerable neurons]. PMID- 16270831 TI - [The Eph receptors and ephrins in synaptic plasticity]. PMID- 16270832 TI - [Progress of BLT2, a new subtype of leukotriene B4 receptors]. PMID- 16270833 TI - [Factors that regulate the expression of resistin gene]. PMID- 16270834 TI - [Study on the receptor proteins and transcription factors related to inflammatory bowel diseases]. PMID- 16270835 TI - [The function of tumor specific antigen (MAGE) in tumor immunotherapy]. PMID- 16270836 TI - [IRAK-4: the key molecule of TLR/IL-1R common signal transduction system]. PMID- 16270837 TI - [A novel acylated peptide--ghrelin]. PMID- 16270838 TI - [Near-field optical microscopy and its application in single biomolecules detection and function studies]. PMID- 16270839 TI - [Proteomics techniques in the investigation of nervous system diseases]. PMID- 16270840 TI - Evaluation of two methods of prompting drivers to use specific exits on conflicts between vehicles at the critical exit. AB - The Florida Department of Transportation used a series of changeable-message signs that functioned as freeway guide signs to divert traffic to Universal Theme Park via one of two eastbound exits based on traffic congestion at the first of the two exits. An examination of crashes along the entire route indicated a statistically significant increase in crashes at the first eastbound exit following the actuation of the system. Furthermore, all of the crashes occurred in close proximity to the exit gore (the crosshatched area at exits that drivers are not supposed to enter or traverse) at the first exit. In Experiment 1, behavioral data were collected using an alternating treatments design. These data revealed that reassigning the exit signs was effective in producing a change in the percentage of drivers using each of the two exits. These data also showed that the reassignment of the theme park exit was associated with an increase in the percentage of motor vehicle conflicts that consisted of vehicles cutting across the exit gore. An analysis revealed that the method used for switching the designated or active theme park exit on the series of changeable-message signs led to the presentation of conflicting messages to some motorists, thus resulting in erratic driving behavior (cutting across the exit gore). In Experiment 2, the treatment evaluated the use of a phased method of switching the designated theme park exit to eliminate the delivery of conflicting messages. The new method for switching the designated theme park exit was not associated with an increase in motorists cutting across the exit gore. PMID- 16270841 TI - A methodology for assessing the functions of emerging speech in children with developmental disabilities. AB - An approach based on Skinner's (1957) theory of verbal behavior has been developed to understand and teach elementary communication skills to children with autism and developmental disabilities (Sundberg & Partington, 1998). However, few studies have directly examined the characteristics of emerging language in children with developmental disabilities. The purpose of this study was to develop and evaluate an assessment for identifying the elementary functions of vocal speech in children. Participants were 4 children with developmental disabilities, aged 6 years to 12 years, who exhibited at least one distinguishable vocal response (word or phrase) frequently in the natural environment. The assessment focused on three verbal operants delineated by Skinner (mand, tact, and intraverbal). One or more functions were identified for at least one vocal response of each child. Results suggested that this assessment would be useful for (a) evaluating Skinner's theory, (b) guiding decisions about language training for individual children, and (c) studying the nature of expressive language development in children with developmental disabilities. PMID- 16270842 TI - Motivational influences on performance maintained by food reinforcement. AB - In Study 1, we examined the independent effects of reinforcer consumption during sessions and meal consumption prior to sessions on performance maintained by food reinforcement. Nine individuals with developmental disabilities participated. On alternate days, a preferred edible item was delivered during (a) seven sessions conducted before lunch (repeated-reinforcement condition) versus (b) one session each conducted before and after lunch (pre- and postmeal conditions). Results for 7 of 9 participants showed decreased response rates across sessions in the repeated-reinforcement condition; results for 3 of 9 participants showed decreased rates during postmeal relative to premeal conditions. Two participants who did not show a decrement in responding during either comparison participated in Study 2, in which reinforcer consumption during sessions, combined with meal consumption prior to sessions, also had no effect on their performance. In Study 3, we determined whether (a) choice of reinforcers, (b) increased break time between sessions, (c) varied reinforcers, or (d) intermittent reinforcement schedules mitigated the satiation effects observed for the 7 participants in Study 1. Presession choice of reinforcers resulted in maintained performance for 2 of 6 participants exposed to this condition. Varied reinforcement resulted in maintained performance for only 1 of 5 participants exposed to this condition. Neither the increased break between sessions nor the intermittent reinforcement schedule was effective in maintaining performance for the participants who were exposed to these conditions. PMID- 16270843 TI - A preliminary analysis of adaptive responding under open and closed economies. AB - In the current investigation, we evaluated the effects of open and closed economies on the adaptive behavior of 2 individuals with developmental disabilities. Across both types of economy, progressive-ratio (PR) schedules were used in which the number of responses required to obtain reinforcement increased as the session progressed. In closed-economy sessions, participants were able to obtain reinforcement only through interaction with the PR schedule requirements (i.e., more work resulted in more reinforcer access). In open-economy sessions, participants obtained reinforcers by responding on the PR schedule and were given supplemental (free) access to the reinforcers after completion of the session. In general, more responding was associated with the closed economy. PMID- 16270845 TI - The impact of enhanced incentives on vocational rehabilitation outcomes for dually diagnosed veterans. AB - This study evaluated the efficacy of adding contingency management techniques to vocational rehabilitation (VR) to improve treatment outcome as measured by entry into competitive employment. Nineteen dually diagnosed veterans who entered VR in the Veterans' Administration's compensated work therapy (CWT) program were randomly assigned to CWT (n = 8) or to CWT with enhanced incentives (n = 11). Over the first 16 weeks of rehabilitation, those in the incentives condition could earn up to dollar 1,006 in cash for meeting two sets of clinical goals: (a) remaining abstinent from drugs and alcohol and (b) taking steps to obtain and maintain a competitive job. Results indicate that relative to participants in the CWT-only group, those in the incentives condition engaged in more job-search activities, were more likely to remain abstinent from drugs and alcohol, were more likely to obtain competitive employment, and earned an average of 68% more in wages. These results suggest that rehabilitation outcomes may be enhanced by restructuring traditional work-for-pay contingencies to include direct financial rewards for meeting clinical goals. PMID- 16270844 TI - Effects of an Internet-based voucher reinforcement program for smoking abstinence: a feasibility study. AB - The present study tested the feasibility of an Internet-based method to obtain objective evidence of smoking abstinence and to deliver vouchers for evidence of abstinence. Four heavy smokers participated in this 4-week study. Twice daily, participants made video recordings of themselves providing a breath carbon monoxide (CO) sample with a Web camera. The video was sent electronically to the smoking clinic. Participants could earn vouchers for gradual reductions in breath CO during an initial shaping condition, and then for achieving abstinence (CO < or = 4 ppm). Vouchers could be exchanged for merchandise at select Internet vendors. Relative to baseline conditions, participants substantially reduced their smoke intake, and 3 achieved sustained periods of abstinence. The study suggests that an Internet-based voucher reinforcement program is a feasible method to promote abstinence from cigarette smoking. PMID- 16270846 TI - Descriptive analyses of caregiver reprimands. AB - We conducted descriptive observations of 5 individuals with developmental disabilities and severe problem behavior while they interacted with their caregivers in either simulated environments (an inpatient hospital facility) or in their homes. The focus of the study was on caregiver reprimands and child problem behavior. Thus, we compared the frequency of problem behavior that immediately preceded a caregiver reprimand to that immediately following a caregiver reprimand, and the results showed that the frequency of problem behavior decreased following a reprimand. It is possible that caregiver reprimands are negatively reinforced by the momentary attenuation of problem behavior, and the implications for long- and short-term effects on caregiver behavior are discussed. PMID- 16270848 TI - Teaching safety skills to children to prevent gun play: an evaluation of in situ training. AB - This study evaluated behavioral skills training with added in situ training for teaching safety skills to prevent gun play. Following baseline, each child received two sessions of behavioral skills training and one in situ training session. Additional in situ training sessions were conducted until the child exhibited the safety skills (don't touch the gun, get away, and tell an adult). All children acquired and maintained the safety skills at a 3-month follow-up. In addition, of the 7 children assessed in a dyad situation, all exhibited the correct skills in the presence of another child. PMID- 16270847 TI - Combined antecedent variables as motivating operations within functional analyses. AB - Functional analysis test conditions typically manipulate a single antecedent variable and an associated consequence to better isolate response-reinforcer relations. In some instances no problem behavior is observed, perhaps representing a false-negative finding. The present study evaluated one approach to assess potentially false-negative findings within functional analyses. Participants were exposed to single-antecedent functional analysis test conditions and combined-antecedent test conditions within a multielement design. Both participants engaged in problem behavior primarily during the combined antecedent test conditions, and treatments matched to the results were effective in reducing problem behavior. Findings are discussed in terms of clinical implications of combining antecedent variables to further examine potentially false-negative functional analysis results. PMID- 16270849 TI - An evaluation of a brief multiple-stimulus preference assessment with adolescents with emotional-behavioral disorders in an educational setting. AB - Brief multiple-stimulus-without-replacement (MSWO) preference assessments were conducted with 3 adolescent boys with emotional-behavioral disorders in the context of their public school educational program. The reinforcing effects of stimuli identified as high, medium, and low preference were then evaluated using an alternating treatments design in which, following an initial baseline, stimuli were delivered contingent on on-task behavior. High-preference stimuli produced the highest percentages of on-task behavior for all 3 participants. PMID- 16270850 TI - An evaluation of simultaneous presentation and differential reinforcement with response cost to reduce packing. AB - We evaluated the effects of multiple treatment procedures, including simultaneous presentation of preferred foods, on the packing behavior of a 9-year-old girl with autism. A reversal design was used to assess the effects of differential reinforcement with response cost alone and with simultaneous presentation. In addition, simultaneous presentation was assessed independent of differential reinforcement with response cost. Results indicated that simultaneous presentation reduced packing and that differential reinforcement with response cost was not necessary to maintain these reductions. Results are discussed in terms of the use of simultaneous presentation for packing as an alternative to consequence manipulations. PMID- 16270851 TI - Preintervention analysis and improvement of customer greeting in a restaurant. AB - We examined customer greeting by employees at one location of a sandwich restaurant chain. First, a preintervention analysis was conducted to determine the conditions under which greeting a customer within 3 s of his or her entry into the restaurant did and did not occur. Results suggested that an appropriate customer greeting was most likely to occur when a door chime was used to indicate that a customer had entered the store and when the store manager was present behind the service counter. Next, a performance improvement intervention, which consisted of the combination of the use of a door chime and manager presence, was evaluated. Results showed that during baseline, a mean of 6% of customers were greeted; during intervention a mean of 63% of customers were greeted. The addition of manager-delivered verbal and graphic group feedback resulted in 100% of customers being greeted across two consecutive sessions. PMID- 16270853 TI - A 19th century predecessor of the token economy. PMID- 16270852 TI - Leisure items as controls in the attention condition of functional analyses. AB - Leisure items (e.g., games, toys) are commonly made available as controls during attention conditions of functional analyses (Ringdahl, Winborn, Andelman, & Kitsukawa, 2002). However, Ringdahl et al. raised questions about this practice. This paper reviews research that supports and conflicts with the inclusion of leisure items as controls, including a quantitative analysis of relevant articles published in the Journal of Applied Behavior Analysis over a 10-year period. Data reviewed suggest that practitioners may consider omitting leisure items as controls or including such items strategically based on the accumulation of assessment information. PMID- 16270854 TI - Special issue on haptics, virtual, and augmented reality. PMID- 16270855 TI - TORNADO: omnistereo video imaging with rotating optics. AB - One of the key techniques for vision-based communication is omnidirectional stereo (omnistereo) imaging, in which stereoscopic images for an arbitrary horizontal direction are captured and presented according to the viewing direction of the observer. Although omnistereo models have been surveyed in several studies, few omnistereo sensors have actually been implemented. In this paper, a practical method for capturing omnistereo video sequences using rotating optics is proposed and evaluated. The rotating optics system consists of prism sheets, circular or linear polarizing films, and a hyperboloidal mirror. This system has two different modes of operation with regard to the separation of images for the left and right eyes. In the high-speed shutter mode, images are separated using postimage processing, while, in the low-speed shutter mode, the image separation is completed by optics. By capturing actual images, we confirmed the effectiveness of the methods. PMID- 16270856 TI - Development of anthropomorphic multi-D.O.F. master-slave arm for mutual telexistence. AB - We developed a robotic arm for a master-slave system to support "mutual telexistence," which realizes remote dexterous manipulation tasks and close physical communication with other people using gestures. In this paper, we describe the specifications of the experimental setup of the master-slave arm to demonstrate the feasibility of the mutual telexistence concept. We developed the master arm of a telexistence robot for interpersonal communication. The last degree of the 7-degree-of-freedom slave arm is resolved by placing a small orientation sensor on the operators arm. This master arm is made light and impedance control is applied in order to grant the operator as much freedom of movement as possible. For this development stage, we compared three control methods and confirmed that the impedance control method is the most appropriate to this system. PMID- 16270857 TI - Data streaming in telepresence environments. AB - In this paper, we discuss data transmission in telepresence environments for collaborative virtual reality applications. We analyze data streams in the context of networked virtual environments and classify them according to their traffic characteristics. Special emphasis is put on geometry-enhanced (3D) video. We review architectures for real-time 3D video pipelines and derive theoretical bounds on the minimal system latency as a function of the transmission and processing delays. Furthermore, we discuss bandwidth issues of differential update coding for 3D video. In our telepresence system-the blue-c-we use a point based 3D video technology which allows for differentially encoded 3D representations of human users. While we discuss the considerations which lead to the design of our three-stage 3D video pipeline, we also elucidate some critical implementation details regarding decoupling of acquisition, processing and rendering frame rates, and audio/video synchronization. Finally, we demonstrate the communication and networking features of the blue-c system in its full deployment. We show how the system can possibly be controlled to face processing or networking bottlenecks by adapting the multiple system components like audio, application data, and 3D video. PMID- 16270858 TI - Dynamic interactions in physically realistic collaborative virtual environments. AB - This work describes our efforts in creating a general object interaction framework for dynamic collaborative virtual environments. Furthermore, we increase the realism of the interactive world by using a rigid body simulator to calculate all actor and object movements. The main idea behind our interactive platform is to construct a virtual world using only objects that contain their own interaction information. As a result, the object interactions are application independent and only a single scheme is required to handle all interactions in the virtual world. In order to have more dynamic interactions, we also created a new and efficient way for human users to dynamically interact within virtual worlds through their avatar. In particular, we show how inverse kinematics can be used to increase the interaction possibilities and realism in collaborative virtual environments. This results in a higher feeling of presence for connected users and allows for easy, on-the-fly creation of new interactions. For the distribution of both the interactive objects and the dynamic avatar interactions, we keep the network load as low as possible. To demonstrate the effectiveness of our techniques, we incorporate them into an existing CVE framework. PMID- 16270859 TI - Six degree-of-freedom haptic rendering using spatialized normal cone search. AB - This paper describes a haptic rendering algorithm for arbitrary polygonal models using a six degree-of-freedom haptic interface. The algorithm supports activities such as virtual prototyping of complex polygonal models and adding haptic interaction to virtual environments. The underlying collision system computes local extrema in distance between the model controlled by the haptic device and the rest of the scene. The haptic rendering computes forces and torques on the moving model based on these local extrema. The system is demonstrated on models with tens of thousands of triangles and developed in an accessibility application for finding collision-free paths. PMID- 16270860 TI - Cutting on triangle mesh: local model-based haptic display for dental preparation surgery simulation. AB - A new method to realize stable and realistic cutting simulation using an impedance display haptic device and microcomputer is presented in this paper. Material removal or cutting simulation is a critical task in dental preparation surgery simulation. In this paper, a piecewise contact force model is proposed to approximately describe the cutting process. Challenging issues of minimizing the difference between the cutting simulation and haptic contact simulation are analyzed. The proposed contact-based simulation method is developed for a one dimensional cutting task and can be expanded to three-dimensional cases. Local model-based multirate simulation cutting architecture is proposed and force control of the haptic device is decoupled from the cutting simulation loop, which can both ensure high fidelity of dynamical simulation as well as maintain stability of the haptic device. The cutting operation is realized using spherical and cylindrical shaped tools. An experiment based on the Phantom desktop proves that fidelity in one-dimensional cutting can be realized and stability in three dimensional cutting can be ensured using the force-filtering method. PMID- 16270861 TI - Psychophysical evaluation of in-situ ultrasound visualization. AB - We present a novel psychophysical method for evaluating ultrasonography based on Real-Time Tomographic Reflection (RTTR), in comparison to Conventional Ultrasound (CUS). The method measures the user's perception of the location of an ultrasound imaged target independently from assessing the action employed to reach it. Three experiments were conducted with the Sonic Flashlight (SF), an RTTR device, and CUS. The first two experiments determined subjects' perception of target location with a triangulation-by-pointing task. Depth perception with the SF was comparable to direct vision, while CUS caused considerable underestimation of target depth. Binocular depth information in the SF was shown to significantly contribute to its superiority. The third experiment tested subjects in an ultrasound-guided needle insertion task. Because the SF provides visualization of the target at its actual location, subjects performed insertions faster and more accurately by using the SF rather than CUS. Furthermore, the trajectory analysis showed that insertions with the SF generally went directly to the target along the desired path, while CUS often led to a large deviation from the correct path consistent with the observed underestimation of target depth. These findings lend great promise to the use of RTTR-based imaging in clinical practice and provide precise means of assessing efficacy. PMID- 16270863 TI - Real-time 3D human capture system for mixed-reality art and entertainment. AB - A real-time system for capturing humans in 3D and placing them into a mixed reality environment is presented in this paper. The subject is captured by nine cameras surrounding her. Looking through a head-mounted-display with a camera in front pointing at a marker, the user can see the 3D image of this subject overlaid onto a mixed reality scene. The 3D images of the subject viewed from this viewpoint are constructed using a robust and fast shape-from-silhouette algorithm. The paper also presents several techniques to produce good quality and speed up the whole system. The frame rate of our system is around 25 fps using only standard Intel processor-based personal computers. Besides a remote live 3D conferencing and collaborating system, we also describe an application of the system in art and entertainment, named Magic Land, which is a mixed reality environment where captured avatars of human and 3D computer generated virtual animations can form an interactive story and play with each other. This system demonstrates many technologies in human computer interaction: mixed reality, tangible interaction, and 3D communication. The result of the user study not only emphasizes the benefits, but also addresses some issues of these technologies. PMID- 16270862 TI - Comparison of path visualizations and cognitive measures relative to travel technique in a virtual environment. AB - We describe a between-subjects experiment that compared four different methods of travel and their effect on cognition and paths taken in an immersive virtual environment (IVE). Participants answered a set of questions based on Crook's condensation of Bloom's taxonomy that assessed their cognition of the IVE with respect to knowledge, understanding and application, and higher mental processes. Participants also drew a sketch map of the IVE and the objects within it. The users' sense of presence was measured using the Steed-Usoh-Slater Presence Questionnaire. The participants' position and head orientation were automatically logged during their exposure to the virtual environment. These logs were later used to create visualizations of the paths taken. Path analysis, such as exploring the overlaid path visualizations and dwell data information, revealed further differences among the travel techniques. Our results suggest that, for applications where problem solving and evaluation of information is important or where opportunity to train is minimal, then having a large tracked space so that the participant can walk around the virtual environment provides benefits over common virtual travel techniques. PMID- 16270864 TI - An infrastructure for realizing custom-tailored augmented reality user interfaces. AB - Augmented Reality (AR) technologies are rapidly expanding into new application areas. However, the development of AR user interfaces and appropriate interaction techniques remains a complex and time-consuming task. Starting from scratch is more common than building upon existing solutions. Furthermore, adaptation is difficult, often resulting in poor quality and limited flexibility with regard to user requirements. In order to overcome these problems, we introduce an infrastructure for supporting the development of specific AR interaction techniques and their adaptation to individual user needs. Our approach is threefold: a flexible AR framework providing independence from particular input devices and rendering platforms, an interaction prototyping mechanism allowing for fast prototyping of new interaction techniques, and a high-level user interface description, extending user interface descriptions into the domain of AR. The general usability and applicability of the approach is demonstrated by means of three example AR projects. PMID- 16270865 TI - Achieving dialogue with children with severe autism in an adaptive multisensory interaction: the "MEDIAte" project. AB - This paper presents an adaptive physical environment that allows children with severe autism to successfully interact with multimodal stimuli, giving them a sense of control of the interaction and, hence, providing them with a sense of agency. This has been an extremely important effort for two main reasons: 1) This user group cannot be typified, hence making the design of an interactive system to fit all the spectrum of individuals a very complex task; 2) each individual PAS (Person on the Autistic Spectrum) user must be able to develop himself within the environment according to his own capacities and potentiality. Qualitative evaluation by psychologists shows very good results and sketches an encouraging future for research on these environments. PMID- 16270866 TI - A particle system for interactive visualization of 3D flows. AB - We present a particle system for interactive visualization of steady 3D flow fields on uniform grids. For the amount of particles we target, particle integration needs to be accelerated and the transfer of these sets for rendering must be avoided. To fulfill these requirements, we exploit features of recent graphics accelerators to advect particles in the graphics processing unit (GPU), saving particle positions in graphics memory, and then sending these positions through the GPU again to obtain images in the frame buffer. This approach allows for interactive streaming and rendering of millions of particles and it enables virtual exploration of high resolution fields in a way similar to real-world experiments. The ability to display the dynamics of large particle sets using visualization options like shaded points or oriented texture splats provides an effective means for visual flow analysis that is far beyond existing solutions. For each particle, flow quantities like vorticity magnitude and wavelength2 are computed and displayed. Built upon a previously published GPU implementation of a sorting network, visibility sorting of transparent particles is implemented. To provide additional visual cues, the GPU constructs and displays visualization geometry like particle lines and stream ribbons. PMID- 16270867 TI - Non-Euclidean spring embedders. AB - We present a conceptually simple approach to generalizing force-directed methods for graph layout from Euclidean geometry to Riemannian geometries. Unlike previous work on non-Euclidean force-directed methods, ours is not limited to special classes of graphs, but can be applied to arbitrary graphs. The method relies on extending the Euclidean notions of distance, angle, and force interactions to smooth non-Euclidean geometries via projections to and from appropriately chosen tangent spaces. In particular, we formally describe the calculations needed to extend such algorithms to hyperbolic and spherical geometries. We also study the theoretical and practical considerations that arise when working with non-Euclidean geometries. PMID- 16270868 TI - [Apoptosis and apoptosis-related genes in experimental autoimmune inner ear disease]. AB - OBJECTIVE: To investigate the protein and mRNA expression patterns of apoptosis related genes, together with evidence of apoptosis, in relation to experimental autoimmune inner ear disease (AIED). METHODS: Male C57BL/6 mice at 4 weeks age (n = 80) were randomly assigned to one of the five group (n = 16). The inbred mice were given a single subcutaneous injection of diluted solution of pertussis and an emulsion containing equal parts of complete Freund adjuvant (CFA) and inner ear antigens (IEAg) extracted form guinea pig. The animals were sacrificed for inner ear examination at a defined time after the immunization (7, 14, 21 or 28 days). An autoimmune inner ear diseases model was established. Apoptosis was detected by terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate (d-UTP) nick end-laying (TUNEL) method. Using immunohistochemical techniques and reverse transcriptase polymerase chain reaction to clarify the profile of Fas, FasL, and bcl-2. RESULTS: Under normal conditions, no TUNEL positive cell was observed in the cochlea except for a few positive cells in the supporting cells of Corti's organ and macula sacculi. Inner ear antigens administration induced TUNEL-positive reactions in a wide variety of cells such as inner hair cells, supporting cells, stria vascularis and spiral ligament fibrocytes. No positive staining was evident in outer hair cells, spiral ganglion cells and Scarpa's ganglion cells during the whole period. Fas proteins were expressed in a wide range of cells in inner ear. The levels of Fas mRNA were no significant differences between normal and AIED mice. FasL and bcl-2 proteins could be detected in spiral ganglion cells and Scarpa's ganglion cells both in normal and AIED mice. FasL positive cells increased in number in inner ear of AIED mice. bcl-2 positive cells were not detectable in inner hair cells, stria vascularis and spiral ligament both in normal and AIED mice. The mRNA of three kinds of apoptosis-related genes was detectable in the normal and AIED mice. FasL mRNA was expressed at low levels in normal, being maximal at 14 d post inoculation and decreased gradually to steady levels by 2 weeks. The levels of bcl-2 mRNA increased significantly during the period of AIED. CONCLUSION: Apoptosis mediated by Fas/FasL signal system may play a role in the initiation and maintenance of AIED. bcl-2 has a crucial role in the regulation of the process of apoptosis in the inner ear of AIED mice. PMID- 16270869 TI - [Construction of the nucleic vaccine pVVP3L-18HN and its antitumor effect on human laryngeal carcinoma]. AB - OBJECTIVE: Nucleic vaccine of pVVP3IL-18HN expressing apoptin gene, Newcastle disease virus HN gene and IL-18 gene were constructed to observe the combinative antitumor effect of the above three genes. METHODS: Eukaryotic expression plasmid pVVP3IL-18HN was constructed by inserting apoptin gene and fragment comprising fused IL-18HN gene and IRES promoter into the downstream of CMV promoter of vector pVAX1. The expression of inserted gene was identified by RT-PCR, indirect immunofluorescence and Western-blot. The recombinant plasmid was introduced into Hep-2 cells by liposome, then suppression rate of Hep-2 of different time and different quantity was calculated according to MTT results. RESULT: The recombinant plasmid of pVVP3IL-18HN suppressed Hep-2 successfully and its suppression rate was up to 61.9% with 20 microg/ml, incubation of 72 hours. CONCLUSION: The nucleic vaccine constructed pVVP3IL-18HN had antitumor effect on Hep-2. It may can be used to the therapy and research of laryngeal carcinoma. PMID- 16270870 TI - [Repair of pharyngeal fistula with marlex mesh]. PMID- 16270871 TI - [Survivin antisense oligonucleotide induces human Hep-2 cell apoptosis]. AB - OBJECTIVE: Survivin highly overexpresses in the most of human tumors, and it may play an important role in the development of tumor. The aim of this study was to explore the effects of survivin antisense oligonucleotide (ASODN) on the proliferation and the apoptosis of human Hep-2 cell. METHODS: Hep-2 cells were transfected with survivin ASODN mediated by lipofectamine, MTT [3-(4,5 dimethylthiazol-2-yl)-2, 5 diphenyl tetrazolium bromide] method was used to observe the cell growth inhibitory rate, the expressions of survivin mRNA and protein were detected by reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot respectively. Flow cytometry was used to examine cell apoptosis rate. Kinase activity test was used to detect the changes of caspase-3 activity. RESULTS: Survivin ASODN obviously inhibited the cell growth of Hep-2 cells after transfection. After transfected with survivin ASODN the expressions of survivin mRNA and protein of Hep-2 cells were down-regulated, and apoptosis rate was significantly increased. The activity of caspase-3 increased highly in Hep-2 cells transfected with survivin ASODN, which showed time-dependent. CONCLUSIONS: Survivin ASODN could inhibit the proliferation of Hep-2 cell and induced apoptosis through down-regulating the the expressions of survivin mRNA and protein. PMID- 16270872 TI - [Cross-check of caloric test and head shaking nystagmus]. AB - OBJECTIVE: To investigate the manifestation of head shaking nystagmus (HSN) and the relationship between HSN and vestibular bithermal test while cross-checking the HSN and vestibular bithermal test. METHODS: One hundred and twenty-four patients were examined by HSN and vestibular bithermal test. The latency and duration of HSN were observed and the dynamic changes of HSN were also investigated. The results of bithermal test were clarified by the values of canal paresis (CP). RESULTS: There were 3 types of HSN, including 41 monophasic HSN, 11 biphasic HSN and 7 perverted HSN. The latency of monophasic HSN was (2.75 +/- 1.41) ms and the duration of it was (32.16 +/- 20.30) ms; as for the biphasic HSN, the first phase had no latency and the duration was (12.33 +/- 4.42) ms which was shorter than that of the second phase (57.00 +/- 17.19) ms (P < 0.01) and the latency of second phase was (57.00 +/- 17.19) ms. The value of canal paresis in the patients without HSN was lower than that in the patients with HSN. CONCLUSIONS: The existence of HSN was dependent on the loss of the function horizontal canal and also on the stage of vestibular compensation. The variation of duration of HSN among individuals was great. In the acute stage of unilateral vestibular function loss, there appeared to be prominent HSN, and the direction of HSN is the same as that of spontaneous nystagmus. With the development of vestibular compensation, biphasic nystagmus would appear instead of monophasic HSN and the direction of first phase represent the direction of un-lesioned side. After compensation, HSN would not be elicited. However,when HSN existed, it suggested that the value of CP should exceed 25%. Perverted nystagmus pointed to central abnormality. PMID- 16270873 TI - [Endoscopic surgery for nasal septal perforation]. AB - OBJECTIVE: To describe the relevant factors of endoscopic surgery in patients with nasal septal perforation. METHODS: Twenty-three patients with nasal septal perforation were treated under nasal endoscope. Four kinds of reconstruction materials were used to accomplish the closure of perforation: residual osseous septum or temporalis fascia, inverting septal mucoperichondrial flap, autologous connective tissue insert overlaid with mucous flaps and turbinate flap. The reconstructed septum was packed by moist dressing with silicone or plastic splints. RESULTS: Seven patients underwent direct closure. Inverting flap repair for five cases, shifting flap closure for ten cases, and repair with turbinate flap in one case. During the follow-up ranging from four weeks to seven months, the successful reconstruction was achieved in 19 cases (82.6%). The problems in the remaining four cases were: mucosal flap displacement, fascia flap shrank and so caused reperforation, two perforations present with only the larger one repaired, the mucosal flap was smaller in size than the perforation. CONCLUSIONS: Intranasal endoscopic reconstruction surgery is a reasonable management for nasal septal perforation. PMID- 16270874 TI - [Expression of adrenomedullin in the tissue with laryngeal carcinoma]. AB - OBJECTIVE: To observe the expression of adrenomedullin (AM) in the patients with laryngeal carcinoma. METHODS: Two-step immunohistochemistry method was used to examine the expression of AM in the patients with laryngeal carcinoma. Radioimmunoassay was applied to determine the concentration of AM in the laryngeal carcinoma tissues, adjacent laryngeal mucosa of carcinoma tissues and in the plasma of patients and controls. RESULTS: Positive stainings for AM were found in all 21 specimen examined,distributed mainly in the cytoplasm of the laryngeal carcinoma cells. Positive stainings were more stronger in the circumference than in the center of tumor tissue for the highly and moderately differentiated tumors. While the stainings were distributed homogeneously for poorly and moderately differentiated tumors. The concentration of AM in the laryngeal carcinoma tissues (n = 44) and the adjacent mucosa (n = 44) were (49.67 +/- 28.33) pg/ml and (14.71 +/- 7.17) pg/ml (x +/- s) respectively and laryngeal tumor showed much higher concentration of AM than the adjacent mucosa (u = 135.00, P < 0.01). The concentration of AM in patients with laryngeal carcinoma of T2, T3 and T4 stage were (31.52 +/- 15.22), (56.63 +/- 18.51) and (96.12 +/- 18.22) pg/ml (x + s) respectively,and there were statistically significant difference among them. In the N stage, patients with higher stages were found to express significantly higher AM concentration, but there was not statistically significant difference between NO stage and N1 stage. In the M stage,patients with M1 stage were found to express significantly higher AM concentration (u = 31.00, P < 0.01). But there was not statistically significant difference between AM plasma concentration of laryngeal carcinoma patients and that of healthy controls. CONCLUSIONS: The results suggested that high expression of AM in tissues of laryngeal carcinoma was related with the TNM stage of laryngeal carcinoma, AM may play an important role in the development of the laryngeal neoplasma. PMID- 16270875 TI - [Treatment and survival analysis of pyriform sinus cancer]. AB - OBJECTIVE: To evaluate the efficacy of surgery for the patients with pyriform sinus carcinoma and analyze the prognostic factors related to the cancer. METHODS: Between December 1995 and December 2002, 62 patients with pyriform sinus cancer were treated in Zhejiang Tumor Hospital. There were 13 patients staged T1, 17 T2, 12 T3, 20 T4. Four patients received preoperative radiation and 40 patients had post-operative radiation. Among 62 patients, 33 patients were treated by partial laryngectomy, 29 patients were treated by total laryngectomy. RESULTS: The survival rate was calculated with Kaplan-Meier method. The overall 3 and 5-year survival rates were 42.3% and 27.8%, respectively. The 3-year survival rate between partial and total laryngectomy was 51.9% and 29.9%. The 5 year survival rate between partial and total laryngectomy was 39.5% and 11.2% (chi2 = 4.14, P<0.05). Early stage and combined modality therapy were the independent favorable prognostic factors. CONCLUSIONS: Early diagnosis with treatment and combined treatment are the most important factors influencing the survival of patients with pyriform sinus carcinoma. Partial laryngopharyngectomy is a suitable treatment for early and selected advanced pyriform sinus carcinoma with a good function and oncologic outcome. PMID- 16270876 TI - [Surgery on laryngeal carcinoma--retrospective analysis of 205 cases]. AB - OBJECTIVE: To evaluate the results of surgery for laryngeal carcinoma. METHODS: A retrospective analysis of long-term therapeutic effect was made on surgery of 105 patients with laryngeal carcinoma in the past 15 years. There were 197 males, 8 cases of female. Tumor stage were included 3 cases of Tis NOMO, 45 cases of T1NOM0, 1 case of T1N1M0, 88 cases of T2NOM0, 3 cases of T2N1M0, 1 case of T2N2M0, 33 cases of T3NOM0, 7 cases of T3N1M0, 3 cases of T3N2M0, 13 cases of T4NOM0, 6 cases of T4N1M0, 1 case of T4N2M0 and 1 case of T4N3M0. Different kinds of operations were as follows: cordectomy (1 case) or stripping (2 cases) by suspended laryngoscope, laryngofissure (9 cases), vertical partial laryngectomy without tracheotomy (16 cases), vertical partial laryngectomy (25 cases), horizontal laryngectomy (7 cases), Arslan's procedure (15 cases), cricohyoidoepiglottopexy (57 cases), subtotal laryngectomy (16 cases) and total laryngectomy (57 cases). RESULTS: There were only four missing cases among all 205 cases. The 3- and 5-year survival rate for glatt and carcinoma is 91.3% and 84. 7% respectively. The 3- and 5-year survival rate for supraglottic carcinoma is 67.5% and 65.0% respectively. Statistical differences were noted between survival rate of glottic and supraglottic carcinoma and between early and advantage disease. The overall 3- and 5-year survival rate is 84.8% and 79.4% respectively. The rate of surgical preservation of laryngeal function is 72.7%. CONCLUSIONS: The prognosis factors of laryngeal cancer were tumor stage, subsite, neck metastasis. All patients treated with different surgical techniques had fairly good quality of functional results. It is possible to preserve the laryngeal function by proper indication, safe margin, combined radiotherapy without compromising survival. PMID- 16270877 TI - [Differentiated thyroid carcinoma in young people]. AB - OBJECTIVE: To investigate the factors that influence survival of the patients with differentiated thyroid carcinoma in young people and evaluate the efficiency of unilateral lobectomy plus isthmectomy with therapeutic cervical lymph node dissection and postoperative TSH (thyroid stimulating hormone) suppressive therapy. METHODS: One hundred and thirty-one patients under 30 years old with differentiated thyroid carcinoma treated in this hospital (14 cases no more than and 117 cases more than 16 years) from Jan. 1st, 1985 to Dec. 31st, 1997 were retrospectively reviewed. One hundred and twenty-eight patients were received only surgery and TSH suppressive therapy, and 3 patients received chemotherapy or radiotherapy because of the progressive metastasis in necks or mediastina. A multivariate analysis was performed in these patients by the Cox proportional hazard model. RESULTS: The mean follow-time (x +/- s) of all patients were (140.86 +/- 43.76) months, with range from 20 to 229 months; Ninety-eight patients followed more than 10 years. Ten patients died of thyroid cancer. The overall 10-year survival rate was 97.18%. The 10-year survival rate for patients < or = 16 years of age and > 16 years were 75.97% and 96.57% respectively (P = 0. 0006). The 10-year survival rate for women and men were 94.91% and 93.69% respectively (P = 0.5261). The 10-year survival rates of patients with papillary thyroid carcinoma and follicular thyroid carcinoma were 93.77% and 96. 55% respectively (P = 0.8137). For patients with tumor size of < or = 1 cm, 1-4 cm and >4 cm the survival rate was 100.0%, 96.40%, and 80.67% respectively (P = 0. 0589). The 10-year survival rates of patients with or without lymph node metastasis were 88.37% and 100. 0% respectively (P = 0.0313). For patients of with or without distant metastasis, The survival rate was 96.64% or 60.00% (P = 0.0000). The 10-year survival rates with or without recurrence were 86. 67% and 95.48% respectively (P = 0. 5681). Using multivariate analysis, risk factors that independently influence survival were distant metastasis, tumor size and age. CONCLUSIONS: The distant metastasis, tumor size and age at diagnosis were the independent factors influencing survival significantly. The status of lymph node metastasis may have certain effect on the prognosis. Unilateral lobectomy plus isthmectomy with a therapeutic cervical lymph node dissection followed by postoperative TSH suppressive therapy is a favourable model to children and young adults with DTC without distant metastasis, but to the patients with distant metastasis, their prognosis of this therapy model is disappointing. PMID- 16270878 TI - [A case of neurilemoma of larynx]. PMID- 16270879 TI - [Diagnosis and reoperation for thyroid carcinoma]. AB - OBJECTIVE: To analysis the reasons of the reoperation of thyroid neoplasm and the efficiency of the surgery and to explore the diagnosis of thyroid neoplasm in order to choose the rational surgical method. METHODS: Reoperation of thyroid cancers were performed in 128 patients from Oct. 1992 to Oct. 2000. The causes of reoperation were thyroid cancer remnants and persistence of the micro carcinoma The type of reoperation includes: 1. completion of lobectomy and isthmectomy or subtotal thyroidectomy. Radical neck dissection or modified neck dissection were indicated for the neck of lymph node metastasis. 2. completion of lobectomy and isthmectomy and modified neck dissection or selective neck dissection for patient with thyroid micro carcinoma. Preoperative fine needle aspiration (FNA) (10 cases), intra-operative frozen section (FS) (55 cases), FNA and FS (13 cases) were done in 78 patients who first visited our hospital, with pure (solitary) thyroid nodule suspected thyroid carcinoma clinically. The results of the above examinations were compared with postoperative pathological results. RESULTS: For the patients with reoperation, the rate of remained cancer was 68.8% (88/128) which was confirmed by pathological results. The occurring of complication was 23.4% (30/128). Laryngeal recurrent nerve paralysis was accounted for 2. 3% and transient postoperative hypocalcemia occurred in 19.5%. Five patients developed local recurrence and 3 had neck metastasis the following up. The 5-, 10-year survival rates of these patients were 92.0% (101/110) and 86.9% (17/20) respectively. The diagnostic accuracy of preoperative FNA, intra-operative FS, FNA and FS were 90.0%, 87.3% and 92.3% respectively. CONCLUSIONS: The rate of postoperative residual carcinoma in thyroid was relatively higher because of misdiagnosis and inadequate operation. It was necessary to take active and reasonable reoperation. Reoperation added to surgical complications. Increasing preoperative diagnostic accuracy & carrying out standard, adequate surgical treatment are the essence to decrease surgical complication. PMID- 16270880 TI - [A chondroma of nasal cavity and paranasal sinus]. PMID- 16270881 TI - [Cancer and surgical treatment impact the quality of life in patients with head and neck cancer]. AB - OBJECTIVE: To evaluate head and neck cancer and surgical treatment impact the quality of life (QOL). METHODS: In this study, 49 cases of head and neck cancer patients were recruited. Among them, 27 cases were laryngeal cancer, 14 cases were tongue cancer and 8 patients were recurrence of nasal pharyngeal cancer after radical radiotherapy. To demonstrate the cancer in different sites of the head and neck impact QOL of the patients in a different way and cancer impact QOL on the physical well-being, social family well-being, emotional well-being, functional well-being of the patients and quality of life (QOL) changed in different time-point before and after operation, QOL was assessed before surgical treatment and at 1,6 months after operation by means of a performance status scale for head and neck cancer patients (PSS-HN) and the functional assessment of cancer therapy head and neck (FACT-H&N) questionnaire. RESULTS: QOL deteriorated significantly in head and neck cancer patients. Cancer in different sites impact on QOL differently especially in patients with tongue cancer (PSS-HN P = 0.0361, FACT-H&N P = 0.0487). Head and neck cancer impact QOL on the physical well-being, social family well-being, emotional well-being, functional well-being of the patients in FACT-H&N questionnaire especially for emotional well-being domains (F = 2.78, P = 0.0311). The QOL in patients deteriorated by surgical treatment and it could be improved following the time. At the 6 months after operation it nearly reached the same scores that assessed before the operation (PSS-HN t = 2.03, P = 0.1120 FACT-H&N t = 1.03, P = 0.1180). Different surgical approaches and different reconstruction methods have different impact on QOL for patients. Laryngeal cancer patients with partial laryngectomy were 107.20 in FACT-H&N while total laryngectomees were 97. 71 at the 6 months after operation, with statistically difference (t = 3.02, P = 0.0430). Tongue cancer patients without reconstruction were 119. 24 in FACT-H&N while the others with reconstruction were 111.39 at the 6 months after operation (t = 3.00, P = 0.0472). CONCLUSIONS: The QOL in head and neck cancer patients can be assessed by the questionnaire and it can be improved by selecting treatment regimen, surgical approaches and reconstructive methods. PMID- 16270882 TI - [Epidemic survey of the middle and aged women related obstructive sleep apnea hypopnea syndrome in Beijing]. AB - OBJECTIVE: To find out the prevalence and risk factors related obstructive sleep apnea hypopnea syndrome (OSAHS) in Chinese women aged 40 years and older. METHODS Participants living in communities must be of age > or = 40 years and were given questionnaires, which developed a scale of 11 questions. Subjects were divided into three groups by scale scores. Those with higher scores were oversampled (2.4%, 14.8%, 42.1% respectively) and subjects of this community-based sample were recorded in the sleep laboratory to ascertain patients (apnea hypopnea index, AHI > or = 5/h and daytime sleepiness). Contrasting patients with the others as a control group, explored differences in symptoms. Stepwise multiple linear regression was used to determine the principal covariates affecting AHI. RESULTS: 34.3% of 1336 eligible women admitted various degree of snoring and 91.0% of those completed questionnaires. Crombach's alpha coefficient of scale reached 0. 7025. Factor analysis reduced 11 questions of scale to four common factors as we have designed: snoring, apneas, other symptoms, risk factors. Fifty nine subjects experienced polysomnography evaluation. The ratios of patients to controls were 2: 25, 4: 18, 10: 16 from lower scores group to higher. We estimated the prevalence of AHI 5,10,15/h and OSAHS in this population were 41.1%, 24.0%, 17.0% and 11.1%, respectively. As far as the frequency of sleep choking, xerostomia or pharyngoxerosis in the morning, these were significant differences between the OSAHS and non-OSAHS groups. Stepwise multiple linear regression analysis identified serum level follicle-stimulating hormone and BMI as predictors of AHI. CONCLUSIONS: This scale had good validity and reliability. Snoring and OSAHS are very common in Chinese women 40 years of age and older. Women with OSAHS did report sleep choking, xerostomia or pharyngoxerosis in the morning. BMI and sex steroids may all play a role in OSAHS. PMID- 16270883 TI - [Research on the origin and ultrastructure microglia-like cell in SD rat Corti's organ after neomycin ototoxicity]. PMID- 16270884 TI - [Carcinogenesis on the local recurrent laryngeal carcinoma]. PMID- 16270885 TI - [Surgical treatment of chronic otitis media with effusion]. PMID- 16270886 TI - [Hearing reconstruction by primary tympanoplasty based on mastoidectomy]. PMID- 16270887 TI - [Risk factors and re-myringoplasty for perforation after tympanic membrane repairing]. PMID- 16270888 TI - [Endoscope with subsidiary incision access for orbital blowout fracture]. PMID- 16270889 TI - [Identification and management of the maxillary sinus ostium in endoscopic sinus surgery]. PMID- 16270890 TI - [Traumatic bleeding vertebra contrary: report of 2 cases]. PMID- 16270891 TI - [Head and neck reconstruction with the anterolateral thigh flap: report of 2 cases]. PMID- 16270892 TI - [One case of malignant fibrous histiocytoma in the sinus]. PMID- 16270893 TI - [Research progress in nonsyndromic sensorineural hearing loss associated genes]. PMID- 16270894 TI - [Pay attention to the diagnosis and therapy of the syndrome of pharynx abnormal sense]. PMID- 16270895 TI - [Twice rupture of cervical artery: a case report]. PMID- 16270896 TI - Overview movement as an index of anesthetic depth in humans and experimental animals. AB - Anesthesia is used widely in animal research, but there are diverse opinions regarding acceptable anesthetic depth. Excessive anesthesia is associated with increased morbidity and mortality. Traditionally, researchers have been taught that animal movement during surgical and experimental procedures indicates that the animal is 'underanesthetized.' Complex movement, however, can be initiated and propagated within the spinal cord, with little input from supraspinal structures. For example, frogs with high spinal-cord transections still maintain the wiping reflex, whereby the hindlimb can move to the forelimb to wipe away a noxious stimulus. Rats that have been decerebrated can perform complex tasks, such as grooming. Brain-dead humans can have spontaneous movement of the arms, legs, and head. Consistent with these phenomena, emerging evidence suggests that, in anesthetized animals, movement in response to noxious stimulation is abolished primarily via anesthetic action in the spinal cord. When isoflurane, halothane, or thiopental is delivered selectively to the brain circulation in goats, substantially greater anesthetic concentrations in brain are needed to ablate movement, as compared with those required upon delivery of anesthetic to the entire body. Rats that have had a precollicular decerebration require the same isoflurane concentrations to prevent movement as compared to intact rats. Furthermore, data from both humans and animals indicate that memory and awareness are ablated at anesthetic concentrations that are < 50% of those needed to abolish movement. Collectively, these data indicate that animals can be anesthetized at depths that, although they do not abolish movement, still produce unconsciousness and amnesia. PMID- 16270897 TI - Detection of Mycoplasma pulmonis by fluorogenic nuclease polymerase chain reaction analysis. AB - Mycoplasma pulmonis induces persistent infections in laboratory mice and rats and can contaminate biological materials. We developed a fluorogenic nuclease polymerase chain reaction (fnPCR) assay to detect M. pulmonis specifically. Primer and probe sequences for the assay were targeted to 16S rRNA sequences specific to M. pulmonis. The assay consistently detected the equivalent of fewer than 10 copies of template DNA. When evaluated against a panel of 24 species of bacteria, the M. pulmonis assay detected only M. pulmonis isolates. Evaluation of 10-fold serial dilutions of cultured M. pulmonis showed that the M. pulmonis fnPCR assay and culture on Dutch agar had comparable sensitivity in detecting viable M. pulmonis organisms, whereas the mouse antibody production test displayed positive serologic results at dilutions higher than those in which viable organisms could be detected. Finally, the M. pulmonis fnPCR assay was able to detect M. pulmonis DNA in nasopharyngeal wash fluid and trachea, lung, and uterus tissue collected from mice naturally infected with M. pulmonis but did not detect the organism in similar samples collected from uninfected, negative control mice. The M. pulmonis fnPCR assay provides a high-throughput, PCR-based method to detect M. pulmonis in infected rodents and contaminated biological materials. PMID- 16270898 TI - Experimental infection of baboons (Papio cynocephalus anubis) with apathogenic and neurovirulent subtypes of herpesvirus papio 2. AB - Cercopithecine herpesvirus 16 (Herpesvirus papio 2; HVP2) is an alpha-herpesvirus of baboons (Papio spp.) that generally causes minimal to inapparent disease in the natural host species. HVP2 is very closely related genetically and antigenically to Cercopithecine herpesvirus 1 (monkey B virus; BV) of macaques, which is well known for its extreme lethality in nonmacaque species including humans. Preliminary evidence suggests that a mouse model of HVP2 would be an excellent tool for studying zoonotic BV infections. Although the pathogenicity of different BV isolates in mice spans the full range of severity from apathogenic to extremely neurovirulent, testing of multiple HVP2 isolates revealed only two distinct phenotypes in mice regardless of route of inoculation: apathogenic (HVP2ap) and highly neurovirulent (HVP2nv). For the HVP2nv mouse model to truly reflect BV infection in both its natural host and the differential pathogenicity of BV in aberrant host species, HVP2nv should not produce severe disease in its natural host. To test this, juvenile baboons were inoculated with doses of 10(6) or 10(4) plaque-forming units of HVP2ap or HVP2nv by using an oral subdermal inoculation route. Parameters followed included the appearance of lesions, shedding of infectious virus, general health, and the immune response to the infection. Regardless of the inoculum dose used, no differences were noted between the two HVP2 subtypes in baboons in any of the parameters measured. These findings further support the use of the HVP2nv mouse system as a model to elucidate and study the viral determinants associated with cross-species BV neurovirulence. PMID- 16270899 TI - Characterization of immune responses during regression of rabbit oral papillomavirus infections. AB - Rabbit oral papillomavirus (ROPV) is a mucosatropic papillomavirus that causes small benign discrete papillomas within the oral cavity of domestic rabbits. The goal of this study was to characterize the immune cell infiltrate over the course of regression of oral papillomas. ROPV-infected oral tissues were harvested at various time points after infection and analyzed by immunohistochemistry for papilloma morphology, viral capsid proteins, and associated immune infiltrates. The results of this study indicated that the L1 and L2 viral capsid proteins were lost rapidly at a time that coincided with an inflammatory response from the rabbit. This inflammatory response began with a rapid rise in numbers of CD11c+ cells at early regression. CD11c+ cells continued to increase in frequency through mid-regression and remained the most-represented cell through late regression. The initial rise in CD11c+ cells was followed by an infiltrate containing increased numbers of activated T cells, including CD4+ and CD25+ cells, during mid-regression. Mid-regression coincided spatially with a loss of viral capsid stain, suggesting that immune cells or cytokines or both were playing a key role in clearance of the papillomas. CD8+ cells increased at the lowest rate and were at low levels in the papilloma epidermis even at mid regression. All cell types decreased by late regression. CD11c+ and major histocompatibility class II+ cells were the last populations of cells to decrease in number. PMID- 16270900 TI - Duration of clinical effects and absence of referred hyperalgesia after femoral vein cannulation in rats. AB - The goal of this study was to assess the duration of pain-related clinical effects and referred hyperalgesia after surgery in rats. Isoflurane anesthesia with or without femoral vein cannulation was performed (n = 6 per group). Body weight and food and water consumption were monitored daily for 48 h, and tail flick latency was measured twice daily for 24 h after surgery. Water consumption at 24 h after surgery was significantly decreased in the surgical group compared with baseline values and those of the anesthesia group. Body weight change and food consumption showed nonsignificant decreases compared with baseline in both groups 24 h after the procedure. There was a trend toward decreased food consumption after surgery compared with that for the anesthesia-alone group. Tail flick latency was nonsignificantly decreased the afternoon after surgery compared with baseline values or that after anesthesia alone. Tail-flick latency was similar to baseline and between groups 24 h after surgery. All parameters were similar between groups and compared with baseline by 48 h after surgery. Our results show some changes in postsurgical pain-related parameters only during the initial 24-h period after femoral cannulation surgery, but only the change in water consumption was significant. Although this study involved only a small number of animals, our findings suggest that femoral vein cannulation produces a less painful stimulus than that seen in studies assessing these parameters after abdominal surgery. Hyperalgesia from a distant painful stimulus could not be measured in this model by using the tail-flick assay. PMID- 16270901 TI - Noninvasive measures of body fat percentage in male Yucatan swine. AB - The purpose of this study was to assess the use of body circumference, ultrasonography, and serum leptin levels as noninvasive measures to estimate body fat percentage in adult, male, Yucatan swine, which are widely used in biomedical research models. Swine (ages 8 to 15 months) were maintained for 20 weeks: control (n = 7); high-fat, high-cholesterol diet (hyperlipidemic; n = 8); alloxan induced diabetes with high-fat, high-cholesterol diet (diabetic dyslipidemic; n = 7); and diabetic dyslipidemic plus exercise-trained (n = 6). Anesthetized swine were positioned on their dorsum for the following measurements: 1) neck, mid abdomen, and widest abdominal girth circumferences; and 2) neck and mid-abdomen ultrasound measurements. Blood samples were obtained for quantification of serum leptin levels. After euthanasia, the carcass and viscera were separated for chemical composition analysis, which demonstrated a significant increase in carcass and visceral fat in the diabetic dyslipidemic swine compared to controls. Serum leptin levels were also increased in the hyperlipidemic and diabetic dyslipidemic swine. Regression analyses demonstrated a significant correlation between carcass fat, visceral fat, and all of the circumference, ultrasound, and serum leptin measures. In conclusion, the widest abdominal girth circumference was the noninvasive measure with the highest predictive value for estimating carcass and visceral fat in adult, male Yucatan miniature swine. PMID- 16270902 TI - Vascular alterations in response to air desiccation injury in the carotid artery of the C3H/HeJ mouse. AB - The purpose of this study was to develop a model of vascular injury in 8-week-old C3H/HeJ mice (weight, 25 to 30 g) by using air desiccation. The carotid arteries were excised 1 to 8 weeks postinjury and evaluated by Verhoeff's stain and immunocytochemistry. In the first group of mice studied (n = 107), neointimal formation occurred and peaked at Day 14. In addition, medial cell division (measured by bromodeoxyuridine labeling) peaked at Day 3, whereas intimal cell proliferation increased gradually throughout the experimental period of 21 days. In addition, extensive thrombus formation occurred within 3 days after injury. The next experiment involved 124 mice and evaluated the effect of anticoagulants on the neointimal and thrombotic response. Mice received aspirin, heparin, or vehicle-only time-release pellets. Both anticoagulants significantly decreased the neointimal and thrombotic responses. The results of this study validate our animal model as being consistent with the Response to Injury Hypothesis of atherogenesis. PMID- 16270903 TI - Theiler's murine encephalomyelitis virus in nonbarrier rat colonies. AB - Theiler's murine encephalomyelitis virus (TMEV), a member of the genus Cardiovirus, is an enteric pathogen of mice that causes acute encephalomyelitis followed by persistent central nervous system infection with chronic inflammation and demyelination after intracerebral inoculation. Although TMEV is a mouse pathogen, antibodies against TMEV strain GDVII have been detected in conventional rat colonies. Natural infection of rats by Cardiovirus has not yet been described. The purpose of this study was to demonstrate TMEV infection of rat colonies by using serologic assays, reverse transcription-polymerase chain reaction (RT-PCR) analysis, and clinical characterization. Indirect immunofluorescence assay of rat serum samples demonstrated antibodies against TMEV-GDVII in 86.3% of samples analyzed, and 77.2% of the antibody-positive samples had neutralizing antibodies. To determine whether rats can be infected experimentally with TMEV-GDVII, specific pathogen-free newborn mice and rats were inoculated intracerebrally with intestinal suspensions from seropositive rats. Both species showed the typical clinical signs of TMEV infection in mice, which is characterized by flaccid hindlimb paralysis and tremor. RT-PCR in brain tissue of experimentally infected animals detected RNA sequences corresponding to the 5' noncoding region of Cardiovirus known as the 'internal ribosome entry site.' These results suggest that rats can be naturally infected with TMEV and related Cardiovirus. Therefore, continued health monitoring for TMEV infection should be included in rat colonies mainly because these animals are used for various experimental purposes. PMID- 16270904 TI - Successful rederivation of contaminated immunocompetent mice using neonatal transfer with iodine immersion. AB - There is an ongoing need to eradicate intercurrent disease from research mouse colonies. Commonly used surgical methods, however, are expensive and time consuming. The purpose of this study was to determine the percentage of litters that could be rederived from infected mouse colonies by neonatal transfer. We immersed neonatal mice in a dilute iodine solution and transferred them to disease-free foster mothers within 48 h of birth. Donor and foster mothers were evaluated for pathogens by serology and fecal polymerase chain reaction (PCR) assay. Of 55 donor mothers, 100% were positive serologically and 59% were positive by fecal PCR for one or more tested organisms, including mouse hepatitis virus, Theiler's murine encephalomyelitis virus, mouse rotavirus, and Helicobacter hepaticus. At 4 to 6 weeks after neonatal transfer, 95% of foster mothers (which served as sentinels for the transferred pups) tested free of pathogens, the exceptions being one case of mouse parvovirus 1 and two of Helicobacter spp. We suggest that cross-fostering is a viable low-cost method for rederivation of mouse colonies contaminated with pathogens such as mouse hepatitis virus, Theiler's murine encephalomyelitis virus, mouse rotavirus, and H. hepaticus. PMID- 16270905 TI - Clinical linoleic acid deficiency in Dahl salt-sensitive (SS/Jr) rats. AB - Male SS/Jr rats were placed on a specially formulated, high-cholesterol, low sodium diet at 3 weeks of age. Of the 50 animals on the diet, 40 developed skin lesions ranging from focal areas of alopecia to diffuse areas of moist dermatitis on the head, face, ear pinnae, and neck. Similar lesions were noted later in 17 of 36 SS/Jr rats in a second study group. Histopathologic findings from two affected animals revealed diffuse, hyperplastic, ulcerative dermatitis, with bacterial colonies of cocci in superficial crusts, as well as chronic hepatic inflammation with hepatocellular glycogen and sinusoidal macrophage aggregates suggestive of lipidosis. Results of a fatty-acid profile of the affected rats showed serum linoleic acid levels of 931 to 1566 micromol/liter, whereas those for control (SS/Jr) samples ranged from 2711 to 3145 micromol/liter. Dietary analysis of the specially formulated diet showed that it contained only 0.225% linoleic acid, which is below the recommended level of 0.3 to 0.6%. In light of the clinical and dietary findings, a diagnosis of linoleic acid deficiency was made. The food manufacturer revised its dietary formulation to increase the linoleic acid content to 1.05%, and no further cases of dermatitis developed in any subsequent groups of rats maintained under the same study protocol. PMID- 16270906 TI - A streamlined method for systematic, high resolution in situ analysis of mRNA distribution in plants. AB - BACKGROUND: In situ hybridisation can provide cellular, and in some cases sub cellular, resolution of mRNA levels within multicellular organisms and is widely used to provide spatial and temporal information on gene expression. However, standard protocols are complex and laborious to implement, restricting analysis to one or a few genes at any one time. Whole-mount and reverse transcriptase-PCR (RT-PCR) based protocols increase throughput, but can compromise both specificity and resolution. With the advent of genome-wide analysis of gene expression, there is an urgent need to develop high-throughput in situ methods that also provide high resolution. RESULTS: Here we describe the development of a method for performing high-throughput in situ hybridisations that retains both the high resolution and the specificity of the best manual versions. This refined semi automated protocol has the potential for determining the spatial and temporal expression patterns of hundreds of genes in parallel on a variety of tissues. We show how tissue sections can be organized on microscope slides in a manner that allows the screening of multiple probes on each slide. Slide handling, hybridisation and processing steps have been streamlined providing a capacity of at least 200 probes per week (depending on the tissue type). The technique can be applied easily to different species and tissue types, and we illustrate this with wheat seed and Arabidopsis floral meristems, siliques and seedlings. CONCLUSION: The approach has the high specificity and high resolution of previous in situ methods while allowing for the analysis of several genes expression patterns in parallel. This method has the potential to provide an analysis of gene expression patterns at the genome level. PMID- 16270907 TI - No effect of omeprazole on pH of exhaled breath condensate in cough associated with gastro-oesophageal reflux. AB - BACKGROUND: Endogenous airway acidification evaluated as pH in exhaled breath condensate (EBC) has been described in patients with chronic cough. Proton pump inhibitors improve gastro-oesophageal reflux (GOR)-associated cough. METHODS: We examined pH levels in EBC and capsaicin cough response in 13 patients with chronic cough (mean age 41 years, SD 9) associated with GOR before and after omeprazole treatment (40 mg/day for 14 days) and its relationship with clinical response. RESULTS: Omeprazole abolished symptoms associated with GOR. Patients with chronic cough had an EBC pH of 8.28 (SD 0.13) prior to treatment but this did not change with omeprazole treatment. There was a significant improvement in the Leicester Cough Questionnaire symptom scores from 80.8 points (SD 13.2) to 95.1 (SD 17) (p = 0.02) and in a 6-point scale of cough scores, but there was no change in capsaicin cough response. CONCLUSION: An improvement in GOR-associated cough was not associated with changes in EBC pH or capsaicin cough response. These parameters are not useful markers of therapeutic response. PMID- 16270908 TI - Palatal development of preterm and low birthweight infants compared to term infants - What do we know? Part 1: The palate of the term newborn. AB - BACKGROUND: The evidence on prematurity as 'a priori' a risk for palatal disturbances that increase the need for orthodontic or orthognathic treatment is still weak. Further well-designed clinical studies are needed. The objective of this review is to provide a fundamental analysis of methodologies, confounding factors, and outcomes of studies on palatal development. One focus of this review is the analysis of studies on the palate of the term newborn, since knowing what is 'normal' is a precondition of being able to assess abnormalities. METHODS: A search profile based on Cochrane search strategies applied to 10 medical databases was used to identify existing studies. Articles, mainly those published before 1960, were identified from hand searches in textbooks, encyclopedias, reference lists and bibliographies. Sources in English, German, and French of more than a century were included. Data for term infants were recalculated if particular information about weight, length, or maturity was given. The extracted values, especially those from non-English paper sources, were provided unfiltered for comparison. RESULTS: The search strategy yielded 182 articles, of which 155 articles remained for final analysis. Morphology of the term newborn's palate was of great interest in the first half of the last century. Two general methodologies were used to assess palatal morphology: visual and metrical descriptions. Most of the studies on term infants suffer from lack of reliability tests. The groove system was recognized as the distinctive feature of the infant palate. The shape of the palate of the term infant may vary considerably, both visually and metrically. Gender, race, mode of delivery, and nasal deformities were identified as causes contributing to altered palatal morphology. Until today, anatomical features of the newborn's palate are subject to a non-uniform nomenclature. CONCLUSION: Today's knowledge of a newborn's 'normal' palatal morphology is based on non-standardized and limited methodologies for measuring a three-dimensional shape. This shortcoming increases bias and is the reason for contradictory research results, especially if pathologic conditions like syndromes or prematurity are involved. Adequate measurement techniques are needed and the 'normal palatal morphology' should be defined prior to new clinical studies on palatal development. PMID- 16270909 TI - Palatal development of preterm and low birthweight infants compared to term infants - What do we know? Part 2: The palate of the preterm/low birthweight infant. AB - BACKGROUND: Well-designed clinical studies on the palatal development in preterm and low birthweight infants are desirable because the literature is characterized by contradictory results. It could be shown that knowledge about 'normal' palatal development is still weak as well (Part 1). The objective of this review is therefore to contribute a fundamental analysis of methodologies, confounding factors, and outcomes of studies on palatal development in preterm and low birthweight infants. METHODS: An electronic literature search as well as hand searches were performed based on Cochrane search strategies including sources of more than a century in English, German, and French. Original data were recalculated from studies which primarily dealt with both preterm and term infants. The extracted data, especially those from non-English paper sources, were provided unfiltered for comparison. RESULTS: Seventy-eight out of 155 included articles were analyzed for palatal morphology of preterm infants. Intubation, feeding tubes, feeding mode, tube characteristics, restriction of oral functions, kind of diet, cranial form and birthweight were seen as causes contributing to altered palatal morphology. Changes associated with intubation concern length, depth, width, asymmetry, crossbite, and contour of the palate. The phenomenon 'grooving' has also been described as a complication associated with oral intubation. However, this phenomenon suffers from lack of a clear-cut definition. Head flattening, pressure from the oral tube, pathologic or impaired tongue function, and broadening of the alveolar ridges adjacent to the tube have been raised as causes of 'grooving'. Metrically, the palates of intubated preterm infants remain narrower, which has been examined up to the age of the late mixed dentition. CONCLUSION: There is no evidence that would justify the exclusion of any of the raised causes contributing to palatal alteration. Thus, early orthodontic and logopedic control of formerly orally intubated preterm infants is recommended, as opposed to non-intubated infants. From the orthodontic point of view, nasal intubation should be favored. The role that palatal protection plates and pressure-dispersing pads for the head have in palatal development remains unclear. PMID- 16270911 TI - Knowledge and use of medicinal plants by local specialists in an region of Atlantic Forest in the state of Pernambuco (Northeastern Brazil). AB - The study of local knowledge about natural resources is becoming increasingly important in defining strategies and actions for conservation or recuperation of residual forests. This study therefore sought to: collect information from local populations concerning the use of Atlantic Forest medicinal plants; verify the sources of medicinal plants used; determine the relative importance of the species surveyed, and; calculate the informant consensus factor in relation to medicinal plant use. Data was obtained using semi-structured forms to record the interviewee's personal information and topics related to the medicinal use of specific plants. The material collected represent 125 plants, distributed among 61 botanical families, with little participation of native plants. This study demonstrated that local people tend to agree with each other in terms of the plants used to treat blood-related problems, but cite a much more diverse group of plants to treat problems related to the respiratory and digestive systems - two important categories in studies undertaken in different parts of the world. The local medicinal flora is largely based on plants that are either cultivated or obtained from anthropogenic zones, possibly due to the use and access restrictions of the legally protected neighboring forest. Despite these restrictions, the species with the highest use-value by this community was Pithecellobium cochliocarpum (Gomez) Macb., a native plant of the Atlantic Forest. PMID- 16270910 TI - Combining metal oxide affinity chromatography (MOAC) and selective mass spectrometry for robust identification of in vivo protein phosphorylation sites. AB - BACKGROUND: Protein phosphorylation is accepted as a major regulatory pathway in plants. More than 1000 protein kinases are predicted in the Arabidopsis proteome, however, only a few studies look systematically for in vivo protein phosphorylation sites. Owing to the low stoichiometry and low abundance of phosphorylated proteins, phosphorylation site identification using mass spectrometry imposes difficulties. Moreover, the often observed poor quality of mass spectra derived from phosphopeptides results frequently in uncertain database hits. Thus, several lines of evidence have to be combined for a precise phosphorylation site identification strategy. RESULTS: Here, a strategy is presented that combines enrichment of phosphoproteins using a technique termed metaloxide affinity chromatography (MOAC) and selective ion trap mass spectrometry. The complete approach involves (i) enrichment of proteins with low phosphorylation stoichiometry out of complex mixtures using MOAC, (ii) gel separation and detection of phosphorylation using specific fluorescence staining (confirmation of enrichment), (iii) identification of phosphoprotein candidates out of the SDS-PAGE using liquid chromatography coupled to mass spectrometry, and (iv) identification of phosphorylation sites of these enriched proteins using automatic detection of H3PO4 neutral loss peaks and data-dependent MS3 fragmentation of the corresponding MS2-fragment. The utility of this approach is demonstrated by the identification of phosphorylation sites in Arabidopsis thaliana seed proteins. Regulatory importance of the identified sites is indicated by conservation of the detected sites in gene families such as ribosomal proteins and sterol dehydrogenases. To demonstrate further the wide applicability of MOAC, phosphoproteins were enriched from Chlamydomonas reinhardtii cell cultures. CONCLUSION: A novel phosphoprotein enrichment procedure MOAC was applied to seed proteins of A. thaliana and to proteins extracted from C. reinhardtii. Thus, the method can easily be adapted to suit the sample of interest since it is inexpensive and the components needed are widely available. Reproducibility of the approach was tested by monitoring phosphorylation sites on specific proteins from seeds and C. reinhardtii in duplicate experiments. The whole process is proposed as a strategy adaptable to other plant tissues providing high confidence in the identification of phosphoproteins and their corresponding phosphorylation sites. PMID- 16270913 TI - Environmental perception of gatherers of the crab 'caranguejo-uca' (Ucides cordatus, Decapoda, Brachyura) affecting their collection attitudes. PMID- 16270912 TI - Palatal development of preterm and low birthweight infants compared to term infants -- What do we know? Part 3: discussion and conclusion. AB - BACKGROUND: It has been hypothesized that prematurity and adjunctive neonatal care is 'a priori' a risk for disturbances of palatal and orofacial development which increases the need for later orthodontic or orthognathic treatment. As results on late consequences of prematurity are consistently contradictory, the necessity exists for a fundamental analysis of existing methodologies, confounding factors, and outcomes of studies on palatal development in preterm and low birthweight infants. METHOD: A search of the literature was conducted based on Cochrane search strategies including sources in English, German, and French. Original data were recalculated from studies which primarily dealt with both preterm and term infants. The extracted data, especially those from non English paper sources, were provided unfiltered in tables for comparison (Parts 1 and 2). RESULTS: Morphology assessment of the infant palate is subject to non standardized visual and metrical measurements. Most methodologies are inadequate for measuring a three-dimensional shape. Several confounding factors were identified as causes contributing to disturbances of palatal and orofacial development. CONCLUSION: Taking into account the abovementioned shortcomings, the following conclusions may be drawn for practitioners and prospective investigators of clinical studies. 1) The lack of uniformity in the anatomical nomenclature of the infant's palate underlines the need for a uniform definition. 2) Metrically, non-intubated preterm infants do not exhibit different palatal width or height compared to matched term infants up to the corrected age of three months. Beyond that age, no data on the subject are currently available. 3) Oral intubation does not invariably alter palatal morphology of preterm and low birthweight infants. 4) The findings on palatal grooving, height, and asymmetry as a consequence of orotracheal intubation up to the age of 11 years are inconsistent. 5) Metrically, the palates of orally intubated infants remain narrower posteriorly, beginning at the second deciduous molar, until the age of 11 years. Beyond that age, no data on the subject are currently available. 6) There is a definite need for further, especially metrical, longitudinal and controlled trials on palatal morphology of preterm and low birthweight infants with reliable measuring techniques. 7) None of the raised confounding factors for developmental disturbances may be excluded until evident results are presented. Thus, early orthodontic and logopedic control of formerly premature infants is recommended up to the late mixed dentition stage. PMID- 16270914 TI - Role of traditional healers in psychosocial support in caring for the orphans: a case of Dar-es Salaam City, Tanzania. AB - Orphans are an increasing problem in developing countries particularly in Africa; due to the HIV/AIDS pandemic; and needs collective effort in intervention processes by including all stakeholders right from the grass roots level. This paper attempts to present the role of traditional healers in psychosocial support for orphan children in Dar-es-Salaam City with special focus on those whose parents have died because of HIV/AIDS. Six traditional healers who were involved in taking care of orphans were visited at their "vilinge" (traditional clinics). In total they had 72 orphans, 31 being boys and 41 being girls with age range from 3 years to 19. It was learned that traditional healers, besides providing remedies for illnesses/diseases of orphans, they also provided other basic needs. Further, they even provided psychosocial support allowing children to cope with orphan hood life with ease. Traditional healers are living within communities at the grass roots level; and appear unnoticed hidden forces, which are involved in taking care of orphans. This role of traditional healers in taking care of orphans needs to be recognised and even scaling it up by empowering them both in financial terms and training in basic skills of psychosocial techniques in how to handle orphans, in order to reduce discrimination and stigmatisation in the communities where they live. PMID- 16270915 TI - Cough: meeting the needs of a growing field. AB - There has been a rapidly increasing volume of research undertaken in the field of acute and chronic cough at both basic scientific and clinical levels. However, until now there has been no journal dedicated to publishing work in this field. In this editorial, we introduce the new online, open-access journal entitled Cough which has been founded specifically for this purpose. We also review the clinical problems posed by acute and chronic cough and highlight some of the current issues that are being tackled by cough researchers. PMID- 16270916 TI - Adenomatoid odontogenic tumor of the mandible: review of the literature and report of a rare case. AB - Adenomatoid odontogenic tumor (AOT) is a rare odontogenic tumor which is often misdiagnosed as odontogenic cyst. To acquire additional information about AOT, all reports regarding AOT and cited in "pubmed" since 1990 onward were reviewed. AOT accounts for about 1% until 9% of all odontogenic tumors. It is predominantly found in young and female patients, located more often in the maxilla in most cases associated with an uneruppted permanent tooth. For radiological diagnose the intraoral periapical radiograph seems to be more useful than panoramic. However, AOT frequently resemble other odontogenic lesions such as dentigerous cysts or ameloblastoma. Immunohistochemically AOT is characterized by positive reactions with certain cytokeratins. Treatment is conservative and the prognosis is excellent. For illustration a rare case of an AOT in the mandible is presented. PMID- 16270917 TI - Metabolic profiling of laser microdissected vascular bundles of Arabidopsis thaliana. AB - BACKGROUND: Laser microdissection is a useful tool for collecting tissue-specific samples or even single cells from animal and plant tissue sections. This technique has been successfully employed to study cell type-specific expression at the RNA, and more recently also at the protein level. However, metabolites were not amenable to analysis after laser microdissection, due to the procedures routinely applied for sample preparation. Using standard tissue fixation and embedding protocols to prepare histological sections, metabolites are either efficiently extracted by dehydrating solvents, or washed out by embedding agents. RESULTS: In this study, we used cryosectioning as an alternative method that preserves sufficient cellular structure while minimizing metabolite loss by excluding any solute exchange steps. Using this pre-treatment procedure, Arabidopsis thaliana stem sections were prepared for laser microdissection of vascular bundles. Collected samples were subsequently analyzed by gas chromatography-time of flight mass spectrometry (GC-TOF MS) to obtain metabolite profiles. From 100 collected vascular bundles (approximately 5,000 cells), 68 metabolites could be identified. More than half of the identified metabolites could be shown to be enriched or depleted in vascular bundles as compared to the surrounding tissues. CONCLUSION: This study uses the example of vascular bundles to demonstrate for the first time that it is possible to analyze a comprehensive set of metabolites from laser microdissected samples at a tissue-specific level, given that a suitable sample preparation procedure is used. PMID- 16270919 TI - Agricultural, domestic and handicraft folk uses of plants in the Tyrrhenian sector of Basilicata (Italy). AB - BACKGROUND: Research was carried out into agricultural and domestic-handicraft uses in folk traditions in the Tyrrhenian sector of the Basilicata region (southern Italy), as it is typically representative of ethnobotanical applications in the Mediterranean area. From the point of view of furnishing a botanical support for the study of local "material culture" data was collected through field interviews of 49 informants, most of whom were farmers. RESULTS: The taxa cited are 60, belonging to 32 botanical families, of which 18 are employed for agricultural uses and 51 for domestic-handicraft folk uses. Data show a diffuse use of plants for many purposes, both in agricultural (present uses 14%; past uses 1%) and for domestic-handicraft use (present uses 40%; past uses 45%); most of the latter are now in decline. CONCLUSION: 60 data look uncommon or typical of the places studied. Some domestic-handicraft folk uses are typical of southern Italy (e.g. the use of Ampelodesmos mauritanicus for making ties, ropes, torches, baskets or that of Acer neapolitanum for several uses). Other uses (e.g. that of Inula viscosa and Calamintha nepeta for peculiar brooms, and of Origanum heracleoticum for dyeing wool red) are previously unpublished. PMID- 16270921 TI - Head & Face Medicine - a new journal for 'intra-interdisciplinary' science. Why? When? Where? AB - The human head and face is the target structure of a large number of medical disciplines which are subject to a continuing trend in medical science - 'ongoing fragmentation' or, to use a better established term, 'opening up new fields'. An adverse side effect of this trend is the separation of scientists, which contributes to a breakdown in communication. Specialization is necessary, but who is able to recombine the pieces of knowledge gained in different branches of science? Who is able to trace back an effect to its cause through the whole system? What is the instrument that enables scientists to think 'laterally', or across disciplines?To be one of these instruments is the vision of Head & Face Medicine. To induce 'intra-interdisciplinary' thinking of scientists by bringing together the findings achieved by different researchers from various specialties, all exploring the same target structure - the human head and face. Head & Face Medicine's objective is to support scientists in gaining new insights from different views, to recognize patterns, to extract new thoughts, to recombine them and bring new visions to life. Evolving tools like the internet, e publishing, Open Access and open peer review make Head & Face Medicine a cross between a traditional journal and a data stream which can be queried, analyzed and processed with the aim of increasing medical knowledge in the area of head and face medicine. These tools represent several advantages: fast publication, increase of a paper's scientific impact and ethical superiority. Head & Face Medicine looks forward to receiving your contributions. PMID- 16270922 TI - Plant Methods: putting the spotlight on technological innovation in the plant sciences. AB - Plant Methods is a new journal for plant biologists, specialising in the rapid publication of peer-reviewed articles with a focus on technological innovation in the plant sciences. The aim of Plant Methods is to stimulate the development and adoption of new and improved techniques and research tools in plant biology. We hope to promote more consistent standards in the plant sciences, and make readily accessible laboratory and computer-based research tools available to the whole community. This will be achieved by publishing Research articles, Methodology papers and Reviews using the BioMed Central Open Access publishing model. The journal is supported by a prestigious editorial board, whose members all recognise the importance of technological innovation as a driver for basic science. PMID- 16270920 TI - An overview of the sensory receptors regulating cough. AB - The cough reflex represents a primary defensive mechanism for airway protection in a variety of mammalian species. However, excessive and inappropriate coughing can emerge as a primary presenting symptom of many airway diseases. Cough disorders are characterized by a reduction in the threshold for reflex initiation and, as a consequence, the occurrence of cough in response to stimuli that are normally innocuous in nature. The current therapeutic strategies for the treatment of cough disorders are only moderately effective. This undoubtedly relates in part to limitations in our understanding of the neural components comprising the cough reflex pathway. The aim of this review is to provide an overview of current concepts relating to the sensory innervation to the mammalian airways, focusing particularly on the sensory receptors that regulate cough. In addition, the review will highlight particular areas and issues relating to cough neurobiology that are creating controversy in the field. PMID- 16270923 TI - Evaluation of an ambulatory system for the quantification of cough frequency in patients with chronic obstructive pulmonary disease. AB - BACKGROUND: To date, methods used to assess cough have been primarily subjective, and only broadly reflect the impact of chronic cough and/or chronic cough therapies on quality of life. Objective assessment of cough has been attempted, but early techniques were neither ambulatory nor feasible for long-term data collection. We evaluated a novel ambulatory cardio-respiratory monitoring system with an integrated unidirectional, contact microphone, and report here the results from a study of patients with COPD who were videotaped in a quasi controlled environment for 24 continuous hours while wearing the ambulatory system. METHODS: Eight patients with a documented history of COPD with ten or more years of smoking (6 women; age 57.4 +/- 11.8 yrs.; percent predicted FEV1 49.6 +/- 13.7%) who complained of cough were evaluated in a clinical research unit equipped with video and sound recording capabilities. All patients wore the LifeShirt system (LS) while undergoing simultaneous video (with sound) surveillance. Video data were visually inspected and annotated to indicate all cough events. Raw physiologic data records were visually inspected by technicians who remained blinded to the video data. Cough events from LS were analyzed quantitatively with a specialized software algorithm to identify cough. The output of the software algorithm was compared to video records on a per event basis in order to determine the validity of the LS system to detect cough in COPD patients. RESULTS: Video surveillance identified a total of 3,645 coughs, while LS identified 3,363 coughs during the same period. The median cough rate per patient was 21.3 coughs.hr-1 (Range: 10.1 cghs.hr(-1) - 59.9 cghs.hr(-1)). The overall accuracy of the LS system was 99.0%. Overall sensitivity and specificity of LS, when compared to video surveillance, were 0.781 and 0.996, respectively, while positive- and negative-predictive values were 0.846 and 0.994. There was very good agreement between the LS system and video (kappa = 0.807). CONCLUSION: The LS system demonstrated a high level of accuracy and agreement when compared to video surveillance for the measurement of cough in patients with COPD. PMID- 16270924 TI - High throughput, high resolution selection of polymorphic microsatellite loci for multiplex analysis. AB - BACKGROUND: Large-scale genetic profiling, mapping and genetic association studies require access to a series of well-characterised and polymorphic microsatellite markers with distinct and broad allele ranges. Selection of complementary microsatellite markers with non-overlapping allele ranges has historically proved to be a bottleneck in the development of multiplex microsatellite assays. The characterisation process for each microsatellite locus can be laborious and costly given the need for numerous, locus-specific fluorescent primers. RESULTS: Here, we describe a simple and inexpensive approach to select useful microsatellite markers. The system is based on the pooling of multiple unlabelled PCR amplicons and their subsequent ligation into a standard cloning vector. A second round of amplification utilising generic labelled primers targeting the vector and unlabelled locus-specific primers targeting the microsatellite flanking region yield allelic profiles that are representative of all individuals contained within the pool. Suitability of various DNA pool sizes was then tested for this purpose. DNA template pools containing between 8 and 96 individuals were assessed for the determination of allele ranges of individual microsatellite markers across a broad population. This helped resolve the balance between using pools that are large enough to allow the detection of many alleles against the risk of including too many individuals in a pool such that rare alleles are over-diluted and so do not appear in the pooled microsatellite profile. Pools of DNA from 12 individuals allowed the reliable detection of all alleles present in the pool. CONCLUSION: The use of generic vector-specific fluorescent primers and unlabelled locus-specific primers provides a high resolution, rapid and inexpensive approach for the selection of highly polymorphic microsatellite loci that possess non-overlapping allele ranges for use in large-scale multiplex assays. PMID- 16270925 TI - Tissue engineering: a challenge of today's medicine. AB - During the last years, tissue engineering-based therapies have been introduced in clinical practice in the head and face area. The regeneration of complex tissue structures for all sites of the body is envisioned for the future. In the present situation, specialists of the different fields publish excellent research papers in specialised journals. As a result, the scientific community, separated towards distinct sub-specialities, has difficulties in communication. To overcome this problem, the demanding, complex and interdisciplinary aspects of tissue engineering has to be approached from new ways. We have conceptualised Head & Face Medicine therefore as a thematically broad ranged journal, including all disciplines involved in the head and neck area. We hope this journal will attract basic researchers and clinicians who are involved in investigating and applying complex themes (examplified by tissue engineering) in the head and face region and will contribute to a gain in scientific information, communication, and collaboration in order to improve the outcome of patient treatments. PMID- 16270926 TI - Cephalometric norms for the Saudi children living in the western region of Saudi Arabia: a research report. AB - BACKGROUND: Previous studies have established specific cephalometric norms for children with different ethnic backgrounds, showing different facial features for each group. Up till now, there is a paucity of information about the cephalometric features of Saudi children living in the western region of Saudi Arabia, who have distinct social and climatic characteristics. The aim of the present study was to establish cephalometric norms for children living in the western region of Saudi Arabia. METHODS: A total of 62 lateral cephalometric radiographs of Saudis (33 females and 29 males; aged 9-12 years) having good facial proportions and Class I dental occlusion, were traced and analyzed. Using the t-test, the mean value, standard deviation and the range of 20 angular and linear variables were calculated and compared to norms of adult Saudis living in the Western region of Saudi Arabia using the t-test. Male and female groups were also compared using the t- test. RESULTS: Saudi children tend to have a significantly shorter and lower face height, a larger angle of convexity, and more proclined and protruded incisors when compared with adult Saudis (P < 0.05). There were no statistically significant differences between male and female groups. CONCLUSION: Saudi children have distinct cephalometric features, which should be used as a reference in the orthodontic treatment of young Saudi patients. PMID- 16270927 TI - Strain driven fast osseointegration of implants. AB - BACKGROUND: Although the bone's capability of dental implant osseointegration has clinically been utilised as early as in the Gallo-Roman population, the specific mechanisms for the emergence and maintenance of peri-implant bone under functional load have not been identified. Here we show that under immediate loading of specially designed dental implants with masticatory loads, osseointegration is rapidly achieved. METHODS: We examined the bone reaction around non- and immediately loaded dental implants inserted in the mandible of mature minipigs during the presently assumed time for osseointegration. We used threaded conical titanium implants containing a titanium2+ oxide surface, allowing direct bone contact after insertion. The external geometry was designed according to finite element analysis: the calculation showed that physiological amplitudes of strain (500-3,000 ustrain) generated through mastication were homogenously distributed in peri-implant bone. The strain-energy density (SED) rate under assessment of a 1 Hz loading cycle was 150 Jm-3 s-1, peak dislocations were lower then nm. RESULTS: Bone was in direct contact to the implant surface (bone/implant contact rate 90%) from day one of implant insertion, as quantified by undecalcified histological sections. This effect was substantiated by ultrastructural analysis of intimate osteoblast attachment and mature collagen mineralisation at the titanium surface. We detected no loss in the intimate bone/implant bond during the experimental period of either control or experimental animals, indicating that immediate load had no adverse effect on bone structure in peri-implant bone. CONCLUSION: In terms of clinical relevance, the load related bone reaction at the implant interface may in combination with substrate effects be responsible for an immediate osseointegration state. PMID- 16270928 TI - Effect of stroke location on the laryngeal cough reflex and pneumonia risk. AB - BACKGROUND: The purpose of this study was to evaluate the risk of developing pneumonia in acute stroke patients comparing the early anatomical stroke location and laryngeal cough reflex (LCR) testing. METHODS: A prospective study of 818 consecutive acute stroke patients utilizing a reflex cough test (RCT), which assesses the neurological status of the LCR compared to magnetic resonance imaging or computerized tomography for stroke location and subsequent pneumonia outcome. Stroke diagnosis and stroke location were made by a neurologist and clinical radiologist, respectively; both were blinded to the RCT results. RESULTS: Brainstem (p-value < .007) and cerebral strokes (p-value < .005) correlated with the RCT results and pneumonia outcome. Of the 818 patients, 35 (4.3%) developed pneumonia. Of the 736 (90%) patients who had a normal RCT, 26 (3.5%) developed pneumonia, and of the 82 (10%) patients with an abnormal RCT, 9 (11%) developed pneumonia despite preventive interventions (p-value < .005). The RCT had no serious adverse events. CONCLUSION: The RCT acted as a reflex hammer or percussor of the LCR and neurological airway protection and indicated pneumonia risk. Despite stroke location, patients may exhibit "brainstem shock," a global neurological condition involving a transient or permanent impairment of respiratory drive, reticular activating system or LCR. Recovery of these functions may indicate emergence from brainstem shock, and help predict morbidity and mortality outcome. PMID- 16270929 TI - Vascular basis of mucosal color. AB - BACKGROUND: Besides the color of the teeth the color of the alveolar gingiva plays a crucial role in esthetic rehabilitation in dento-alveolar treatment. Whereas nowadays the color of the teeth can be determined exactly and individually, the specific influence of the red color of the gingiva on treatment has not been assessed yet. The aim of this study was to evaluate the vascularization as the basis for gingival esthetics. METHODS: Standardized photographs of defined areas of the alveolar gingiva in operated and non-operated patients were taken and assigned to groups with same characteristics after color comparisons. In addition, histologic and immunohistologic analyses of gingival specimens were performed for qualitative and quantitative assessment of vessels and vascularization. Finally, colors and number of vessels were correlated. RESULTS: Our results demonstrated three different constellations of colors of the alveolar gingiva in healthy patients. The operated patients could not be grouped because of disparate depiction. There was a clear correlation between color and vessel number in the alveolar gingiva. CONCLUSION: Our investigations revealed the connections between vascularization and gingival color. Recommendations for specific change or even selection of colors based on the results cannot be given, but the importance of vascularly based incision lines was demonstrated. PMID- 16270930 TI - Urinary diseases and ethnobotany among pastoral nomads in the Middle East. AB - This article is derived from a broad, twenty-year study of ethnobotany and folk medicine among pastoral nomads in the Middle East which took place from 1984 to 2004. The article presents examples of different treatments of diseases and disorders of the urinary tract carried out by healer herbalists. The preparation of remedies includes boiling infusions, extraction of dry or fresh leaves, flowers, seeds or whole plants. Some of these plants were used both as food and as medicine, by ingesting different parts of the plants, such as leaves, flowers, fruits, and so on, either while soft, cooked or dried. Data were collected by using unstructured interviews and by observation. These plants were identified by healers, patients, and university botanists. This paper identified eighty-five plant species, which belong to thirty-six families. The most representative families are: Asteraceae (8), Brassicaceae (6), Poaceae (6), Umbelliferae (6). PMID- 16270931 TI - Why study the use of animal products in traditional medicines? AB - The World Health Organization (WHO) estimates that as many as 80% of the world's more than six billion people rely primarily on animal and plant-based medicines. The healing of human ailments by using therapeutics based on medicines obtained from animals or ultimately derived from them is known as zootherapy. The phenomenon of zootherapy is marked both by a broad geographical distribution and very deep historical origins. Despite their importance, studies on the therapeutic use of animals and animal parts have been neglected, when compared to plants. This paper discusses some related aspects of the use of animals or parts thereof as medicines, and their implications for ecology, culture (the traditional knowledge), economy, and public health. PMID- 16270932 TI - Change in bronchial responsiveness and cough reflex sensitivity in patients with cough variant asthma: effect of inhaled corticosteroids. AB - BACKGROUND: Cough variant asthma (CVA) is a cause of chronic cough and a precursor of typical asthma. We retrospectively examined the longitudinal change in bronchial responsiveness and cough reflex sensitivity in CVA patients with respect to the effect of long-term inhaled corticosteroids (ICS). METHODS: Provocative concentration of methacholine causing a 20% fall in forced expiratory volume in one second (PC20-FEV1) and provocative concentration of capsaicin eliciting 5 or more coughs (C5) were measured before treatment and during a follow up period following relief of cough (median; 2.0 (range; 0.5 to 8.0) years after the initial visit) in a total of 20 patients with CVA (7 males and 13 females, mean +/- SD age of 49.9 +/- 12.9 years). RESULTS: Three of 8 patients not taking long-term ICS developed typical asthma compared to none of 12 patients taking ICS (p = 0.0171). PC20-FEV1 significantly (p < 0.0001) increased from 1.80 (GSEM, 1.35) to 10.7 (GSEM, 1.63) mg/ml in patients taking ICS but did not change in patients not taking ICS [2.10 (GSEM, 1.47) compared to 2.13 (GSEM, 1.52) mg/ml]. Cough threshold did not change in patients whether taking or not taking ICS. CONCLUSION: Long-term ICS reduces bronchial hyperresponsiveness in CVA as recognized in typical asthma. Cough reflex sensitivity is not involved in the mechanism of cough in CVA. PMID- 16270934 TI - Review of methodologies and a protocol for the Agrobacterium-mediated transformation of wheat. AB - Since the first report of wheat transformation by Agrobacterium tumefaciens in 1997, various factors that influence T-DNA delivery and regeneration in tissue culture have been further investigated and modified. This paper reviews the current methodology literature describing Agrobacterium transformation of wheat and provides a complete protocol that we have developed and used to produce over one hundred transgenic lines in both spring and winter wheat varieties. PMID- 16270933 TI - Comparison of cough reflex sensitivity after an inhaled antigen challenge between actively and passively sensitized guinea pigs. AB - BACKGROUND: Late asthmatic response is observed following antigen challenge in actively, but not passively, sensitized guinea pigs. Although cough reflex sensitivity is increased after antigen challenge in actively sensitized guinea pigs, it is unknown whether the antigen-induced increase in cough reflex sensitivity develops in passively sensitized animals. The aim of this study was to compare the cough reflex sensitivity to inhaled capsaicin after an inhaled antigen challenge between actively and passively sensitized guinea pigs. METHODS: Measurement of number of coughs elicited by increasing concentrations of capsaicin (10(-6) and 10(-4) M) and bronchial responsiveness to ascending concentrations of methacholine, and analysis of bronchoalveolar lavage fluid (BALF) were separately performed 24 h after an antigen challenge in actively and passively sensitized guinea pigs. RESULTS: Percentage of eosinophils in BALF and bronchial responsiveness to methacholine were increased 24 h after the antigen challenge in both actively and passively sensitized animals compared with saline challenged actively and passively sensitized animals, respectively. Absolute number of eosinophils in BALF from actively sensitized and antigen-challenged guinea pigs was significantly greater than that from passively sensitized and antigen-challenged animals. Cough response to capsaicin and concentration of substance P in BALF were increased 24 h after the antigen challenge in actively sensitized guinea pigs, but not in passively sensitized guinea pigs. Bronchial responsiveness, cough reflex sensitivity and substance P concentration and total cells in BALF were increased in actively sensitized and saline challenged guinea pigs compared with passively sensitized and saline challenged animals. CONCLUSION: The results suggest that active sensitization per se increases cough reflex sensitivity accompanied by increased inflammatory cells and substance P level in BALF, and antigen challenge further increases them, while simple IgE- and/or IgG-mediated allergic reaction per se or the low intensity of eosinophil infiltration in the airway itself may not affect cough reflex sensitivity in guinea pigs. PMID- 16270935 TI - A rare cause of specific cough in a child: the importance of following-up children with chronic cough. AB - For many years, the term 'specific cough' has been used as a clinical cough descriptor in children to signify the likelihood of an underlying disease causing the cough. In this case study, we describe a child with specific cough caused by a rare carcinoma, a mucoepidermoid carcinoma of the bronchus. The cough only totally resolved after the primary cause was successfully treated. This report highlights the importance of following up children with cough, especially those with specific cough. PMID- 16270936 TI - Focusing on the ethnobotanical uses of plants in Mersin and Adana provinces (Turkey). AB - This paper presents the result of a study on the herbal drugs in the herbal markets in Mersin and Adana. The data were collected through direct interviews with herbalists and customers between 2002-2005 and the popular medicinal plants were investigated. A total of 107 species belonging to 56 families were investigated and the samples were listed with their local and Latin names. The investigation includes cross-checking the disorders and their herbal cures and their recommended use stated by the local herbalists, by the parts used, and by the preparations. The cultivated species and their ethno botanical uses, are documented and extensive inventory is presented. As a result, we observed that these plants are used especially for intestinal digestive disorders of the gastrointestinal tract, (21.68%), respiratory tract system disorders (10.43%), heart-blood circulatory system disorders (8.48%), urinary tract system disorders (7.70%), skin disorders (6.48%) and others. PMID- 16270938 TI - A rapid and versatile combined DNA/RNA extraction protocol and its application to the analysis of a novel DNA marker set polymorphic between Arabidopsis thaliana ecotypes Col-0 and Landsberg erecta. AB - BACKGROUND: Many established PCR-based approaches in plant molecular biology rely on lengthy and expensive methods for isolation of nucleic acids. Although several rapid DNA isolation protocols are available, they have not been tested for simultaneous RNA isolation for RT-PCR applications. In addition, traditional map based cloning technologies often use ill-proportioned marker regions even when working with the model plant Arabidopsis thaliana, where the availability of the full genome sequence can now be exploited for the creation of a high-density marker systems. RESULTS: We designed a high-density polymorphic marker set between two frequently used ecotypes. This new polymorphic marker set allows size separation of PCR products on agarose gels and provides an initial resolution of 10 cM in linkage mapping experiments, facilitated by a rapid plant nucleic acid extraction protocol using minimal amounts of A. thaliana tissue. Using this extraction protocol, we have also characterized segregating T-DNA insertion mutations. In addition, we have shown that our rapid nucleic acid extraction protocol can also be used for monitoring transcript levels by RT-PCR amplification. Finally we have demonstrated that our nucleic acid isolation method is also suitable for other plant species, such as tobacco and barley. CONCLUSION: To facilitate high-throughput linkage mapping and other genomic applications, our nucleic acid isolation protocol yields sufficient quality of DNA and RNA templates for PCR and RT-PCR reactions, respectively. This new technique requires considerably less time compared to other purification methods, and in combination with a new polymorphic PCR marker set dramatically reduces the workload required for linkage mapping of mutations in A. thaliana utilizing crosses between Col-0 and Landsberg erecta (Ler) ecotypes. PMID- 16270939 TI - Idiopathic chronic cough: a real disease or a failure of diagnosis? AB - Despite extensive diagnostic evaluation and numerous treatment trials, a number of patients remain troubled by a chronic and uncontrollable cough. Eosinophilic bronchitis, atopic cough and non-acid reflux have been recently added to the diagnostic spectrum for chronic cough. In some cases, failure to consider these conditions may explain treatment failure. However, a subset of patients with persisting symptoms may be regarded as having an idiopathic cough. These individuals are most commonly female, of postmenopausal age and frequently report viral upper respiratory tract infections as an initiating event. This paper seeks to explore the validity of idiopathic cough as a distinct clinical entity. PMID- 16270940 TI - Ethnopharmacological survey of different uses of seven medicinal plants from Mali, (West Africa) in the regions Doila, Kolokani and Siby. AB - An ethnopharmacological survey was carried out to collect information on the use of seven medicinal plants in rural areas in the nearby regions of Bamako, Mali. The plants were Opilia celtidifolia, Anthocleista djalonensis, Erythrina senegalensis, Heliotropium indicum, Trichilia emetica, Piliostigma thonningii and Cochlospermum tinctorium. About 50 medical indications were reported for the use of these plants in traditional medicine. The most frequent ailments reported were malaria, abdominal pain and dermatitis. The highest number of usages was reported for the treatment of malaria (22%). The majority of the remedies were prepared from freshly collected plant material from the wild and from a single species only. They were mainly taken orally, but some applications were prepared with a mixture of plants or ingredients such as honey, sugar, salt, ginger and pepper. Decoction of the leaves was the main form of preparation (65%) and leaf powder was mostly used for the preparation of infusions (13%). The part of the plants most frequently used was the leaves. There was a high degree of informant consensus for the species and their medicinal indications between the healers interviewed. The results of this study showed that people are still dependent on medicinal plants in these rural areas of Mali. PMID- 16270941 TI - The ethnobotany of Christ's Thorn Jujube (Ziziphus spina-christi) in Israel. AB - This article surveys the ethnobotany of Ziziphus spina-christi (L.) Desf. in the Middle East from various aspects: historical, religious, philological, literary, linguistic, as well as pharmacological, among Muslims, Jews, and Christians. It is suggested that this is the only tree species considered "holy" by Muslims (all the individuals of the species are sanctified by religion) in addition to its status as "sacred tree " (particular trees which are venerated due to historical or magical events related to them, regardless of their botanical identity) in the Middle East. It has also a special status as "blessed tree" among the Druze. PMID- 16270937 TI - Cough: are children really different to adults? AB - Worldwide paediatricians advocate that children should be managed differently from adults. In this article, similarities and differences between children and adults related to cough are presented. Physiologically, the cough pathway is closely linked to the control of breathing (the central respiratory pattern generator). As respiratory control and associated reflexes undergo a maturation process, it is expected that the cough would likewise undergo developmental stages as well. Clinically, the 'big three' causes of chronic cough in adults (asthma, post-nasal drip and gastroesophageal reflux) are far less common causes of chronic cough in children. This has been repeatedly shown by different groups in both clinical and epidemiological studies. Therapeutically, some medications used empirically for cough in adults have little role in paediatrics. For example, anti-histamines (in particular H1 antagonists) recommended as a front line empirical treatment of chronic cough in adults have no effect in paediatric cough. Instead it is associated with adverse reactions and toxicity. Similarly, codeine and its derivatives used widely for cough in adults are not efficacious in children and are contraindicated in young children. Corticosteroids, the other front-line empirical therapy recommended for adults, are also minimally (if at all) efficacious for treating non-specific cough in children. In summary, current data support that management guidelines for paediatric cough should be different to those in adults as the aetiological factors and treatment in children significantly differ to those in adults. PMID- 16270942 TI - Trends and characteristics of oral and maxillofacial injuries in Nigeria: a review of the literature. AB - BACKGROUND: The etiology of maxillofacial injuries varies from one country to another and even within the same country depending on the prevailing socioeconomic, cultural and environmental factors. Periodic verification of the etiology of maxillofacial injuries helps to recommend ways in which maxillofacial injuries can be averted. The aim of the present study is therefore to analyse the characteristics and trends of maxillofacial injuries in Nigeria based on a systematic review of the literature. METHODS: A literature search using MEDLINE was conducted for publications on maxillofacial injuries in Nigeria. The relevant references in these publications were manually searched for additional non Medline articles or abstracts. Forty-two studies met the inclusion criteria and the full-texts of these articles were thoroughly examined. Due to lack of uniformity and consistency in assessment and measurement variables, and treatment modalities in most of the studies, it was impossible to apply the traditional methods of a systematic review. Therefore, a narrative approach was conducted to report the findings of the included studies. RESULTS: Although, other causes like assaults, sport injuries, and industrial accidents increased in numbers, throughout the period between 1965 and 2003, road traffic crashes remained the major etiological factor of maxillofacial injuries in all regions, except northeastern region where assault was the major cause. A significant increase in motorcycles related maxillofacial injuries was observed in most urban and suburban centres of the country. Animal attacks were not an unusual cause of maxillofacial injuries in most parts of northern Nigeria. Patients in the age group of 21-30 years were mostly involved. A strong tendency toward an equal male to-female ratio was observed between earlier and later periods. CONCLUSION: Road traffic crashes remain the major cause of maxillofacial injuries in Nigeria, unlike in most developed countries where assaults/interpersonal violence has replaced road traffic crashes as the major cause of the injuries. There is a need to reinforce legislation aimed to prevent road traffic crashes and the total enforcement of existing laws to reduce maxillofacial injuries among children and adults. Special attention should also be paid by the authority to improve the socioeconomic conditions of Nigerian populace. PMID- 16270945 TI - Rare-earth iodides in ionic liquids: the crystal structure of [SEt3]3[LnI6] (Ln = Nd, Sm). AB - Crystals of [SEt3]3[LnI6] (Ln = Nd, Sm) were obtained by the reaction of LnI2 with the ionic liquid [SEt3][Tf2N] [Tf2N = bis(trifluoromethanesulfonyl)imide]. The compounds are characterized by octahedral [LnI6]3- units that are surrounded by a distorted cube of triethylsulfonium cations. PMID- 16270944 TI - Protocol: precision engineering of plant gene loci by homologous recombination cloning in Escherichia coli. AB - Plant genome sequence data now provide opportunities to conduct molecular genetic studies at the level of the whole gene locus and above. Such studies will be greatly facilitated by adopting and developing further the new generation of genetic engineering tools, based on homologous recombination cloning in Escherichia coli, which are free from the constraints imposed by the availability of suitably positioned restriction sites. Here we describe the basis for homologous recombination cloning in E. coli, the available tools and resources, together with a protocol for long range cloning and manipulation of an Arabidopsis thaliana gene locus, to create constructs co-ordinately driven by locus-specific regulatory elements. PMID- 16270943 TI - Cell type-specific characterization of nuclear DNA contents within complex tissues and organs. AB - BACKGROUND: Eukaryotic organisms are defined by the presence of a nucleus, which encloses the chromosomal DNA, and is characterized by its DNA content (C-value). Complex eukaryotic organisms contain organs and tissues that comprise interspersions of different cell types, within which polysomaty, endoreduplication, and cell cycle arrest is frequently observed. Little is known about the distribution of C-values across different cell types within these organs and tissues. RESULTS: We have developed, and describe here, a method to precisely define the C-value status within any specific cell type within complex organs and tissues of plants. We illustrate the application of this method to Arabidopsis thaliana, specifically focusing on the different cell types found within the root. CONCLUSION: The method accurately and conveniently charts C value within specific cell types, and provides novel insight into developmental processes. The method is, in principle, applicable to any transformable organism, including mammals, within which cell type specificity of regulation of endoreduplication, of polysomaty, and of cell cycle arrest is suspected. PMID- 16270946 TI - Crystal structure of stoichiometric YBa2Fe3O8. AB - The oxygen-nonstoichiometry locus in YBa2Fe3O(8+w) changes with the sign of w. The singularity at w = 0 is studied by Rietveld refinement of combined high resolution, high-intensity neutron and synchrotron X-ray diffraction data collected at room temperature on red-colored stoichiometric YBa2Fe3O8. A residual disorder of oxygen atoms is identified in a concentration of 0.03 vacancy/interstitials per formula, with corresponding anisotropy in thermal displacements of oxygen neighbors. The disorder improves accommodation of trivalent iron over the two unequal coordination polyhedra and is not identical with the charged Frenkel disorder induced thermally. The symmetry of the cooperative magnetic order is Ib'am, with opposite moments of equal magnitude (4.07 micro(B) at 295 K) at the two iron sites. PMID- 16270947 TI - Structure and reactivity of a pyridine-1-imido-2-thiolato complex of iridium(III), CpIr(1-N-2-Spy), generated by photolysis of the (azido)(pyridine-2 thiolato) complex, CpIr(2-Spy)(N3). AB - Photolysis of the (azido)(pyridine-2-thiolato)iridium(III) complex CpIr(2 Spy)(N3) (1) gave a pyridine-1-imido-2-thiolato complex, CpIr(1-N-2-Spy) (2), in which one of the nitrogen atoms of the azide ligand has been inserted into the Ir N(py) bond (Cp = eta5-C5Me5). Complex 2 reacted quantitatively with methyl iodide to give the N-methylated product, [CpIr(1-NMe-2-Spy)]I (3). X-ray crystallography revealed that both 2 and 3 have similar two-legged piano stool structures with planar 1-N-2-Spy2- or 1-NMe-2-Spy- ligands, which form iridacyclopentadienyl-like rings by moderate S(ppi)/N(ppi) to Ir(dpi) pi donation. PMID- 16270948 TI - Self-assembly of a nonanuclear nickel(II) complex and its magnetic properties. AB - A nonanuclear nickel(II) complex [Ni9(PyA)(10)(mu3-OH)2(mu2-OH)2(mu2 OH2)2(H2O)6](ClO4)4.12H2O, containing pyridine-2-aldoximato monoanion, PyA, has been structurally and magnetochemically characterized. The nickel(II) ions, linked together via several bi- or trifurcated oximato, hydroxo, and water bridges, are antiferromagnetically coupled with the coupling constants J1 = -26.5 cm(-1) and J2 = -7.0 cm(-1), resulting in a ground state of S(t) = 1. PMID- 16270949 TI - A novel open-framework copper borophosphate: Cu(H2O)2[B2P2O8(OH)2]. AB - A new open-framework copper borophosphate was hydrothermally synthesized and structurally characterized. Its structure may be regarded as a layer forming via vertex-sharing BO3(OH) and PO4 tetrahedra bonded together with CuO2(OH)2(H2O)2 octahedral units. The thermal stability and magnetic properties are also discussed. PMID- 16270950 TI - Cationic five-coordinate Pt(II) complexes as donors in the formation of Pt-->Ag dative bonds. AB - A unique trinuclear cluster containing two short Pt-->Ag dative bonds was prepared and characterized crystallographically. Two monocationic orthometalated Pt(II) complexes containing 2-phenylpyridine and the tridentate ligand 9S3 (1,4,7 trithiacyclononane) exhibit axial sulfur interactions with Pt as well as short dative bonds. The Pt-->Ag dative bonding results in a dramatic shortening of the axial Pt-S distance in each Pt complex cation, relative to the mononuclear Pt complex. Evidence for Pt-Ag interactions in solution is also presented. PMID- 16270951 TI - Oxygenation of a ruthenium(II) thiolate to a ruthenium(II) sulfinate proceeds via ruthenium(III). AB - Exposure of acetonitrile/methanol solutions of [PPN][Ru(DPPBT)3] [PPN = bis(triphenylphosphoranylidene); DPPBT = 2-diphenylphosphinobenzene thiolate] to oxygen initiates metal-centered oxidation, yielding the ruthenium(III) thiolate Ru(DPPBT)3. Ru(DPPBT)3 further reacts with oxygen, at sulfur, to give the ruthenium(III) sulfinate complex [Ru(DPPBT-O2)2(DPPBT)], which is reduced under ambient conditions to [PPN][Ru(DPPBT-O2)2(DPPBT)]. Ruthenium(II) sulfinate is the only product isolated from acetonitrile/methanol. Yellow crystals of [PPN][Ru(DPPBT-O2)2(DPPBT)] were obtained. Ruthenium(III) sulfinate was isolated as green prism-shaped crystals upon oxygenation of [PPN][Ru(DPPBT)3] in chlorobenzene/hexane. Electrochemical oxidation of ruthenium(II) sulfinate yields the ruthenium(III) derivative, which is rapidly reduced back to ruthenium(II) upon the addition of hydroxide. PMID- 16270952 TI - Group IV complexes of a tetradentate amine mono(phenolate) ligand: a second side arm donor stabilizes cationic species. AB - An amine mono(phenolate) ligand bearing two side-arm donors led to octahedral trialkoxo and trialkyl group IV metal complexes, in which one of the donors was unbound, and to exceptionally stable cationic complexes in which the two side-arm donors were tightly bound. PMID- 16270953 TI - Deuterium-induced copper pairing in Zr2CuD(approximately 5). AB - Deuteration of Zr2Cu leads to a reconstruction of its MoSi2-type metal atom arrangement. While the tetragonal alloy contains isolated copper atoms (Cu-Cu = 3.22 angstroms) in a cubic zirconium atom environment, the monoclinic deuteride Zr2CuD4.71 contains Cu2 dumbbells (Cu-Cu = 2.39 angstroms), of which each copper atom has a trigonal-prismatic zirconium atom environment. Deuterium occupies five sites, of which four have tetrahedral metal configurations (two Zr4-type and two Zr3Cu-type) and are fully occupied while one has a trigonal-bipyramidal metal configuration (Zr3Cu2-type) and is partially occupied (71%). Copper is bonded to four deuterium atoms in a saddle-like configuration (Cu-D = 1.73-1.87 angstroms). Two of the ligands connect copper in a nearly linear Cu-D-Cu arrangement to partially interrupted dimeric [Cu2D6](n)() ribbons running perpendicular to the Cu2 dumbbell direction. At the upper phase limit, the ribbons are presumably no longer interrupted and the deuteride can be described by the limiting ionic formula 2Zr2+[Cu(+)D3](2-)2D-. PMID- 16270954 TI - A tetranuclear Cr(III)Ni(II)3 cyano-bridged complex based on M(tacn) derivative building blocks. AB - The reaction of Cr(Bztacn)(CN)3 (Bztacn is 1,4,7-trisbenzyl-1,4,7 triazacyclononane) with Ni(iPrtacn)Cl2 (iPrtacn is 1,4,7-trisisopropyl-1,4,7 triazacyclononane) affords a CrNi3 tetranuclear complex. Variable temperature and magnetization versus field measurements show a S = 9/2 ground state and an appreciable magnetic anisotropy with a negative D(9/2) value equal to -0.54 cm( 1). Magnetization studies on one single crystal using a micro-SQUID show a fast tunneling process at zero field at very low temperature. PMID- 16270955 TI - Hexaazaphenalenyl anion revisited: a highly symmetric planar pi system with multiple-networking ability for self-assembled metal complexation. AB - The hexaazaphenalenyl anion (HAP, 3-), a highly symmetric heterocyclic pi system, has been synthesized and characterized. The crystal structures of its potassium salt K+.3- and copper complex Cu2+.(NH3)4.(3-)2 show pi-pi stacking and radially extended hydrogen bonds as well as coordination bonds constructed by the HAP anion. These in-plane and out-of-plane strong interactions demonstrate the multiple-networking ability of the HAP anion. PMID- 16270957 TI - Chiral supramolecules: organometallic molecular loops made from enantiopure R [cis-Rh2(C6H4PPh2)2(CH3CN)6](BF4)2. AB - Two enantiopure molecular loops, RR-[cis-Rh2(C6H4PPh2)2(py)2O2C(CF2)(n)CO2]2 (1, n = 2 and 2, n = 3) have been made from the reaction in CH2Cl2 and CH3OH of the inherently chiral dirhodium compound, R-[cis-Rh2(C6H4PPh2)2(CH3CN)6](BF4)2, and HO2C(CF2)(n)CO2H in the presence of an excess of pyridine. Single-crystal structure analyses reveal that each of these compounds is composed of two R-cis Rh2(C6H4PPh2)2(ax-py)2(2+) units, and two equatorial perfluorodicarboxylate linkers, which form a loop oligomer. The 1H, 19F, and 31P[1H] NMR spectra in CDCl3 and C5D5N indicate that only one type of highly symmetric species exists in each solution, which is consistent with the solid-state structures. PMID- 16270956 TI - Metal effect on the supramolecular structure, photophysics, and acid-base character of trinuclear pyrazolato coinage metal complexes. AB - Varying the coinage metal in cyclic trinuclear pyrazolate complexes is found to significantly affect the solid-state packing, photophysics, and acid-base properties. The three isoleptic compounds used in this study are [[3,5 (CF3)2Pz]M]3 with M = Cu, Ag, and Au (i.e., Cu3, Ag3, and Au3, respectively). They form isomorphous crystals and exist as trimers featuring nine-membered M3N6 rings with linear two-coordinate metal sites. On the basis of the M-N distances, the covalent radii of two-coordinate Cu(I), Ag(I), and Au(I) were estimated as 1.11, 1.34, and 1.25 angstroms, respectively. The cyclic [[3,5-(CF3)2Pz]M]3 complexes pack as infinite chains of trimers with a greater number of pairwise intertrimer M...M interactions upon proceeding to heavier coinage metals. However, the intertrimer distances are conspicuously short in Ag3 (3.204 angstroms) versus Au3 (3.885 angstroms) or Cu3 (3.813 angstroms) despite the significantly larger covalent radius of Ag(I). Remarkable luminescence properties are found for the three M3 complexes, as manifested by the appearance of multiple unstructured phosphorescence bands whose colors and lifetimes change qualitatively upon varying the coinage metal and temperature. The multiple emissions are assigned to different phosphorescent excimeric states that exhibit enhanced M...M bonding relative to the ground state. The startling luminescence thermochromic changes in crystals of each compound are related to relaxation between the different phosphorescent excimers. The trend in the lowest energy phosphorescence band follows the relative triplet energy of the three M(I) atomic ions. DFT calculations indicate that [[3,5-(R)2Pz]M]3 trimers with R = H or Me are bases with the relative basicity order Ag << Cu < Au while fluorination (R = CF3) renders even the Au trimer acidic. These predictions were substantiated experimentally by the isolation of the first acid-base adduct, [[Au3]2:toluene]infinity, in which a trinuclear Au(I) complex acts as an acid. PMID- 16270959 TI - Chiral organometallic triangles with Rh-Rh bonds. 2. Compounds prepared from enantiopure cis-Rh2(C6H4PPh2)2(OAc)2(HOAc)2 and their catalytic potentials. AB - Enantiomers of the orthometalated dirhodium compound cis Rh2(C6H4PPh2)2(OAc)2(HOAc)2 (R-1 and S-1) were prepared from carboxylate exchange reactions of the resolved diasteroisomers of cis-Rh2(C6H4PPh2)2(protos)2(H2O)2 (protos = N-4-methylphenylsulfonyl-l-proline anion) and acetic acid. These compounds react with excess Me3OBF4 in CH3CN, producing the enantiomers of [cis Rh2(C6H4PPh2)2(CH3CN)6](BF4)2 (R-2 and S-2) which have six labile and replaceable CH3CN ligands in equatorial and axial positions. Reactions of R-2 and S-2 with tetraethylammonium salts of the linear dicarboxylic acids, terephthalic acid (HO2CC6H4CO2H), oxalic acid (HO2CCO2H), and 4,4'-diphenyl-dicarboxylic acid (HO2CC6H4C6H4CO2H) afford the enantiopure triangular supramolecules [cis Rh2(C6H4PPh2)2(O2CC6H4CO2)(py)2]3, RRR-3 and SSS-3, Rh6(cis C6H4PPh2)6(O2CCO2)3(py)5(CH2Cl2), RRR-4 and SSS-4, and Rh6(cis C6H4PPh2)6(O2CC6H4C6H4CO2)3(py)4(CH2Cl2)2, RRR-5 and SSS-5, respectively. The absolute structures of each of the enantiomers of 1, 3, 4, and 5 were determined by X-ray diffraction analyses. The enantiomers of 3, 4, and 5 were found to be enantiomorphically isostructural, whereas R-1 and S-1 crystallized in different space groups. In 4 and 5, CH2Cl2 molecules coordinate to rhodium atoms in the axial positions. The 1H and 31P[1H] NMR spectra of all compounds are reported. The triangular compounds are redox-active, and their electrochemistry is also discussed. An assay of the catalytic activity/selectivity performance of the triangles for typical metal carbene transformation, using the model intermolecular cyclopropanation of styrene with ethyl diazoacetate in both homogeneous and heterogeneous phases, show that these chiral triangles are very active and have remarkable selectivity when compared with simple Rh2 paddle-wheel catalysts with chiral amidate ligands. PMID- 16270960 TI - Structural and spectroscopic features of mono- and binuclear nickel(II) complexes with tetradentate N(amine)2S(thiolate)2 ligation. AB - Three complexes containing Ni(II)N(amine)2S(thiolate)2 units have been prepared and characterized. Both (R,R)-N,N'-bis(1-carboxy-2-mercaptoethyl)-1,2 diaminoethane [(R,R)-1] and N,N'-bis(2-methyl-2-mercaptoprop-1-yl)-1,3-diamino 2,2-dimethylpropane (4) act as tetradentate S-N-N-S ligands to form complexes Ni((R,R)-1) and Ni4 with nearly planar cis-NiN2S2 units. The N-Ni-N and S-Ni-S angles differ significantly in the two complexes yet are very nearly supplementary. The 1,3-disubstituted cyclohexane species rac-N,N'-bis(2-mercapto 2-methylprop-1-yl)-1,3-cyclohexanediamine (6) behaves as a bis(bidentate-N,S) ligand to form an unexpectedly intense-blue dinickel complex (1S,3R,1'S,3'R)-7, which contains two trans-NiN2S2 units bridged by 1,3-disubstituted cyclohexane groups. The coordination geometry in (1S,3R,1'S,3'R)-7 is distorted 15 degrees toward tetrahedral, most likely as a result of steric crowding, suggested by several short contacts between the NiS2 units and both the cyclohexyl and gem dimethyl groups of the N,S-chelate rings. The complexes exhibit rich UV-vis spectra, whose deconvoluted bands are now fully assigned, from low to high energy, as ligand field (LF), pi(S) --> Ni(II) ligand-to-metal charge transfer (LMCT), sigma(S) --> Ni(II) LMCT, sigma(N) --> Ni(II) LMCT, localized S, and S,N Rydberg transitions. The unusually intense LF absorptions shown by (1S,3R,1'S,3'R)-7 are thought to result from relaxation of the Laporte restriction arising from the 15 degrees tetrahedral twist. PMID- 16270958 TI - Oxomolybdenum tetrathiolates with sterically encumbering ligands: modeling the effect of a protein matrix on electronic structure and reduction potentials. AB - The effect of sterically encumbering ligands on the electronic structure of oxomolybdenum tetrathiolate complexes was determined using a combination of electronic absorption and magnetic circular dichroism spectroscopies, complimented by DFT bonding calculations, to understand geometric and electronic structure contributions to reduction potentials. These complexes are rudimentary models for a redox-active metalloenzyme active site in a protein matrix and allow for detailed spectroscopic probing of specific oxomolybdenum-thiolate interactions that are directly relevant to Mo-S(cysteine) bonding in pyranopterin molybdenum enzymes. Data are presented for three para-substituted oxomolybdenum tetrathiolate complexes ([PPh4][MoO(p-SPhCONHCH3)4], [PPh4][MoO(p SPhCONHC(CH2O(CH2)2CN)3)4], and [PPh4][MoO(p-SPhCONHC(CH2O(CH2)2COOCH2CH3)3)4]). The Mo(V/IV) reduction potentials of the complexes in DMF are -1213, -1251, and 1247 mV, respectively. The remarkably similar electronic absorption and magnetic circular dichroism spectra of these complexes establish that the observed reduction potential differences are not a result of significant changes in the electronic structure of the [MoOS4]- cores as a function of the larger ligand size. We provide evidence that these reduction potential differences result from the driving force for a substantial reorganization of the O-Mo-S-C dihedral angle upon reduction, which decreases electron donation from the thiolate sulfurs to the reduced molybdenum center. The energy barrier to favorable O-Mo-S-C geometries results in a reorganizational energy increase, relative to [MoO(SPh)4](-/2-), that correlates with ligand size. The inherent flexible nature of oxomolybdenum-thiolate bonds indicate that thiolate ligand geometry, which controls Mo-S covalency, could affect the redox processes of monooxomolybdenum centers in pyranopterin molybdenum enzymes. PMID- 16270961 TI - Bismuth borates: two new polymorphs of BiB3O6. AB - Two new polymorphs of BiB3O6 were identified at low temperatures using boric acid as a flux. Unlike alpha-BiB3O6, which crystallizes in a noncentrosymmetric space group and, thus, shows exceptional nonlinear optical (NLO) properties, beta BiB3O6 (I) and gamma-BiB3O6 (II) crystallize in the centrosymmetric space group P2(1)/n with the following lattice parameters: a = 14.1664(1), b = 6.7514(1), c = 4.4290(1) angstroms, beta = 102.125(1) degrees for I; and a = 8.4992(1), b = 11.7093(1), c = 4.2596(1) angstroms, beta = 121.141(1) degrees for II. However, from the structural point of view, the three polymorphs of BiB3O6 are closely related. The structure of beta-BiB3O6 (I) contains a 2-dimensional borate layer, which could be considered to be an intralayered additive product of alpha-BiB3O6. On the other hand, the 3-dimensional borate framework in gamma-BiB3O6 (II) could be considered to be an interlayered additive product of beta-BiB3O6 (I). According to the synthesis experiments and calculated density, it is proposed that compounds I and II should be the low-temperature (high-pressure) polymorphs of BiB3O6. PMID- 16270962 TI - Cyclic trimer versus head-tail dimer in metal-nucleobase complexes: importance of relative orientation (syn, anti) of the metal entities and relevance as a metallaazacrown compound. AB - The formation and crystal structure analysis of a cyclic trinuclear Pd complex with bridging 1-methylcytosinato model nucleobases is reported: [[(tmeda)Pd(1 MeC(-)-N3,N4)]3] (ClO4)3.5.5H2O (tmeda = N,N,N',N'-tetramethylethylenediamine; 1 MeC- = 1-methylcytosine deprotonated at exocyclic amino group) is obtained, among others, from the hydroxo-bridged dinuclear species [[(tmeda)Pd(OH)]2](ClO4)2, which likewise has been characterized by X-ray crystallography, and 1-MeC (1-MeC = neutral 1-methylcytosine) in aqueous solution. The usual head-tail dimer (HT1) appears not to be formed presumably because of the steric bulk of the tmeda ligand, which prevents a close approach of two tmeda ligands. There is also no evidence for formation of an alternative head-tail dimer structure (HT2) which, in principle, would not lead to any steric clash of ligands, but would require an orientation of the metal at N4 that is almost perpendicular to the nucleobase plane. In the Pd3 compound, the bridging metals are approximately in an anti arrangement, thereby leading to Pd...Pd separations within the Pd3 triangle close to 5.2 angstroms. This arrangement is reflected in the 1H NMR spectrum by a strongly deshielded H5 resonance of the nucleobase, occurring at 6.56 ppm (D2O). The overall structure of the Pd3 is that of a double cone, with ClO4- counterions approaching the cavities from either side. The trinuclear structure is also maintained in Me2SO-d6. In this solvent, Pd3 acts as a fluoride anion receptor, with F- binding to the N4H protons, as evident from large downfield shifts of these protons. The compound is compared with cyclic adeninato complexes of hexacoordinated metal ions, and a conceptional analogy with [12]metallacrown-3 species is outlined. PMID- 16270963 TI - Structural, electronic, and vibrational characterization of Fe-HNO porphyrinates by density functional theory. AB - A recent report of the structural and vibrational properties of heme-bound HNO in myoglobin, MbHNO, revealed a long Fe-N(HNO) bond with the hydrogen atom bonded to the coordinated N atom. The Fe-N(H)-O moiety was reported to exhibit an unusually high Fe-N(HNO) stretching frequency relative to those of the corresponding [FeNO]6 and [FeNO]7 porphyrinates, despite the Fe-N(HNO) bond being longer than either of its Fe-N(NO) counterparts. Herein, we present results from density functional theory calculations of an active site model of MbHNO that support the previous assignment and clarify this seemingly contradictory result. The results are consistent with the experimental evidence for a ground-state Fe-N(H)-O structure having a long Fe-N(HNO) bond and a uniquely high nu(Fe)(-)(N(HNO)) frequency. This high frequency is the result of the correspondingly low reduced mass of the normal mode, which is largely attributable to significant motion of the N-bound hydrogen atom. Additionally, the calculations show the Fe-N(H)O bonding in this complex to be remarkably insensitive to whether the HNO and ImH ligand planes are parallel or perpendicular. This is attributed to insensitivities of the Fe-L(axial) characters of molecular orbitals to the relative HNO and ImH orientation in both the parallel and perpendicular conformers. PMID- 16270964 TI - Influence of the reaction temperature and ph on the coordination modes of the 1,4 benzenedicarboxylate (BDC) ligand: a case study of the Ni(II)(BDC)/2,2' bipyridine system. AB - Three new Ni(BDC)/2,2'-bipy compounds, Ni2(BDC)(HBDC)2(2,2'-bipy)2 (2), Ni3(BDC)3(2,2'-bipy)2 (3), and Ni(BDC)(2,2-bipy)2.2H2O (5), in addition to the previously reported Ni(BDC)(2,2'-bipy).0.75H2BDC (1) and Ni(BDC)(2,2'-bipy)(H2O) (4) [BDC = 1,4-benzenedicarboxylate; 2,2'-bipy = 2,2'-bipyridine], have been synthesized by hydrothermal reactions. A systematic investigation of the effect of the reaction temperature and pH resulted in a series of compounds with different compositions and dimensionality. The diverse product slate illustrates the marked sensitivity of the structural chemistry of polycarboxylate aromatic ligands to synthesis conditions. Compound 1, which has a channel structure containing guest H2BDC molecules, is formed at the lowest pH. The guest H2BDC molecules are connected by hydrogen bonds and form extended chains. At a slightly higher pH, a dimeric molecular compound 2 is formed with a lower number of protonated carboxylate groups per nickel atom and per BDC ligand. Reactions at higher temperature and the same pH lead to the transformation of 1 and 2 into the two-dimensional, layered trinuclear compound 3. As the pH is increased, a one dimensional polymer 4 is formed with a water molecule coordinated to Ni2+. Bis monodentate and bischelating BDC ligands alternate along the chain to give a crankshaft rather than a regular zigzag arrangement. A further increase of the pH leads to the one-dimensional chain compound 5, which has two chelating 2,2'-bipy ligands. Crystal data: 2, triclinic, space group P, a = 7.4896(9) angstroms, b = 9.912(1) angstroms, c = 13.508(2) angstroms, alpha = 86.390(2) degrees , beta = 75.825(2) degrees, gamma = 79.612(2) degrees, Z = 2; 3, orthorhombic, space group Pbca, a = 9.626(2) angstroms, b = 17.980(3) angstroms, c = 25.131(5) angstroms, Z = 4; 5, orthorhombic, space group Pbcn, a = 14.266(2) angstroms, b = 10.692(2) angstroms, c = 17.171(2) angstroms, Z = 8. PMID- 16270965 TI - Novel metal-linked face-to-face porphyrazine dimer. AB - We report the synthesis and physical studies of a novel porphyrazine (pz) dimer [1[Ni,Cu]]2, which has Ni(II) ions incorporated into the pz cores and is linked by two Cu(II) ions coordinated to bis(picolinamide) chelates attached to the pz periphery. [1[Ni,Cu]]2 was prepared from precursor pz 2 with a selenodiazole ring fused to the pz core. This ring was deprotected to form the diamino-pz 3, which reacted with 2 mol of picolinoyl chloride hydrochloride to form pz 1[2H,2H], with peripheral bis(picolinamide) chelates; this was metalated to form [1[Ni,Cu]]2. The crystal structures of 1[2H,2H] and [1[Ni,Cu]]2 are presented. The latter is a dimer in which parallel, face-to-face pz's with an average separation of 3.30 angstroms are linked through the peripheral picolinamide ligands by a pair of peripheral Cu(II) ions. Each Cu(II) is coordinated with distorted square-planar geometry by a picolinamide from each pz. In this report, we focus on the interaction of these two peripheral Cu(II) ions. We discuss the preparation and magnetic properties of the pz dimer complex [1[Ni,Cu]]2 with two Cu(II) ions in the peripheral chelate but a diamagnetic metal ion Ni(II) in the pz core. Although [1[Ni,Cu]]2 contains two Cu(II) ions (S = 1/2), we could detect no electron paramagnetic resonance signal, which suggests very strong antiferromagnetic exchange between those two Cu(II) ions. Temperature-dependent magnetic susceptibility measurement gives an exchange splitting between the S = 0 ground state and the excited triplet state of delta = 660 cm(-1). PMID- 16270966 TI - Theoretical investigation of paramagnetic diazabutadiene gallium(III)-pnictogen complexes: insights into the interpretation and simulation of electron paramagnetic resonance spectra. AB - The electronic structures and the spin density distributions of the paramagnetic gallium 1,4-diaza(1,3)butadiene (DAB) model systems [((t)Bu DAB)Ga(I)[Pn(SiH3)2]]* and the related dipnictogen species [((t)Bu DAB)Ga[Pn(SiH3)2]2]* (Pn = N, P, As) were studied using density functional theory. The calculations demonstrate that all systems share a qualitatively similar electronic structure and are primarily ligand-centered pi-radicals. The calculated electron paramagnetic resonance (EPR) hyperfine coupling constants (HFCCs) for these model systems were optimized using iterative methods and were used to create accurate spectral simulations of the parent radicals [((t)Bu DAB)Ga(I)[Pn(SiMe3)2]]* (Pn = N, P, or As) and [((t)Bu-DAB)Ga[Pn(SiMe3)2]2]* (Pn = P or As), the EPR spectra of which had not been simulated previously due to their complexity. Excellent agreement was observed between the calculated HFCCs and the optimum values, which can be considered the actual HFCCs for these systems. The computational results also revealed inconsistencies in the published EPR data of some related paramagnetic group 13-DAB complexes. PMID- 16270967 TI - Synthesis, structures, and magnetic properties of heteronuclear Cu(II)-Ln(III) (Ln = La, Gd, or Tb) complexes. AB - Facile one-pot reactions led to the formations of dinuclear [CuLn(hmp)2(NO3)3(H2O)2] (Ln = Tb (1.Tb), Gd (1.Gd), or La (1.La)), and trinuclear [Cu2Ln(mmi)4(NO3)(H2O)2](ClO4)(NO3).2H2O (Ln = Tb (2.Tb) or Gd (2.Gd)) and [Cu2La(mmi)4(NO3)2(H2O)](ClO4).2H2O (2.La) with polydentate ligands 2 (hydroxymethyl)-pyridine and 2-hydroxymethyl-1-methyl-imidazole. In these complexes, each pair of Cu(II) and Ln(III) ions is linked by a double mu-alkoxo bridge. The temperature dependences of the magnetic susceptibilities of 1 and 2 were investigated in the range of 2-300 K. The dinuclear and trinuclear Cu-Gd complexes exhibit ferromagnetic interaction. The coupling constant J values in the heterodinuclear Cu-Gd complexes are correlated to values of the dihedral angles alpha between the two O-Cu-O and O-Gd-O fragments of the bridging CuO2Gd networks, with the largest J value associated with the smallest alpha value. The occurrence of a ferromagnetic interaction between Cu(II) and Gd(III) ions of the trinuclear entity is supported by the field dependence of the magnetization. The field dependence of the magnetization at 2 K of 1.Gd and 2.Gd confirms the nature of the ground state and of the Cu(II)-Gd(III) interaction, while alternating current susceptibility measurements demonstrates out-of-phase ac susceptibility signals of 1.Tb, which is the molecule-based magnetic material of the smallest nuclearity which exhibits frequency-dependent behavior within the 3d-4f mixed metal systems. PMID- 16270968 TI - Bimodal system (luminophore and paramagnetic contrastophore) derived from Ln(III) complexes based on a bipyridine-containing macrocyclic ligand. AB - The synthesis of a new 15-membered polyaza-macrocyclic ligand L3H3, which is based on a 2,2'-bipyridine moiety and a diethylenetriaminetriacetic acid core, is reported. The lanthanide chelates of this octadentate ligand were programmed for bimodal probes, luminescent agents (Sm, Eu, Tb, Dy), and magnetic resonance imaging agents (Gd3+). The neutral 1:1 complexes with these Ln3+ ions were prepared and studied in aqueous solution by luminescence and NMR techniques. The main photophysical characteristics of these complexes (i.e., the absorption and luminescence spectra, the metal-centered lifetimes, and the overall luminescence yields, Phi) were measured. In addition, the role played by nonradiative pathways (vibrational energy transfer involving coordinated water molecules, involvement of ligand-to-metal charge-transfer excited states, or metal --> ligand back transfer) is discussed. The L3.Eu and L3.Tb complexes show very bright luminescence when photoexcited from the lowest-energy absorption band of the bipyridine chromophore. The luminescence quantum yields in an air-equilibrated water solution at room temperature are 0.10 and 0.21, respectively, despite the presence of one water molecule in the first coordination sphere of the metal ion. NMR data show that L3.Gd contains also one H2O molecule in the inner sphere. The proton longitudinal relaxivity, r1, of this complex is 3.4 s(-1) mM(-1) (0.47 T, 310 K) and the rotational correlation time, tau(R), is 57 ps (310 K). These values are comparable to those of the clinically used Gd-DTPA. Interestingly, the water exchange rate between the coordination site and the bulk solvent is slow (tau(M) = 3.5 micros at 310 K). The presence of water molecules in the second sphere and in rapid exchange with the solvent is discussed. Finally, it was found by luminescence and NMR experiments that these lanthanide complexes are stable versus transmetalation by several cations (especially Ca2+ and Zn2+) at physiological pH and have no interaction with blood proteins. PMID- 16270969 TI - High-spin diimine complexes of iron(II) reject binding of carbon monoxide: theoretical analysis of thermodynamic factors inhibiting or favoring spin crossover. AB - A new series of Fe(II) complexes, FeCl2[N(R)=C(Me)C(Me)=N(R)], containing diimine ligands with hemilabile sidearms R (R = CH2(CH2)2NMe2, 1, CH2(CH2)2OMe, 2, CH2(CH2)2SMe), 3) were synthesized. The crystal structure of 1 showed 6 coordination where both amine arms were attached, whereas 2 was a 5-coordinate 16e species with one methoxy arm dangling free. Extensive attempts were made to bind CO to these species to synthesize precursors for dihydrogen complexes but were unsuccessful. Reaction of 1 with 1 or 2 equiv of AgOTf under CO atmosphere resulted in isolation of only a 6-coordinate bis(triflate)-containing product [Fe[N(R)=C(Me)C(Me)=N(R)](OTf)2] (R = CH2(CH2)2NMe2), 5. Reaction of 5-coordinate 2 with AgSbF6 under CO did not give a CO adduct but afforded instead a dicationic dinuclear complex [Fe[N(R)=C(Me)C(Me)=N(R)](mu-Cl)]2[SbF6]2 (R = CH2(CH2)2OMe), 4, containing a weakly bound SbF6. Thus coordination of hard-donor anions to iron was favored over CO binding. The unexpected rejection of binding of CO is rationalized by the iron being in a high-spin state in this system and energetically incapable of spin crossover to a low-spin state. Theoretical calculations on CO interaction with Fe(II) centers in spin states S = 0, 1, and 2 for both the 16e complexes and their CO adducts aid further understanding of this problem. They show that interaction of CO with a high-spin 5-coordinate Fe model diimine complex is essentially thermoneutral but is exergonic by about 48 kcal/mol to a comparable but low-spin diphosphine fragment. Spin crossover is thus disfavored thermodynamically rather than kinetically (e.g. a "spin block" effect); i.e., the ligand field strengths of the primarily N-donor groups are apparently insufficient to give a low-spin CO adduct. PMID- 16270970 TI - Comparative study of ruthenium(II) tris(bipyridine) derivatives for electrochemiluminescence application. AB - Several [Ru(bpy)3]2+ (bpy = 2,2'-bipyridine and its derivatives) complexes were synthesized and compared electrochemically and spectroscopically in the search for better luminophores for electrochemiluminescence (ECL)-based analytical applications. ECL measurement in [Ru(bpy)3]2+/tripropylamine (TPA) aqueous buffer solutions has led to a conclusion that due to the complexity of the ECL generation process, the photoluminescence efficiency cannot be used to predict ECL intensity and there is no obvious relationship between the photoluminescence quantum yield and the ECL intensity. Under the present experimental condition, when compared with the pristine [Ru(bpy)3]2+, the ethoxycarbonyl-substituted derivative, [Ru(bpy-COOEt)3]2+, one of the most efficient luminophores under photoexcitation, did not generate reasonably intense ECL, whereas luminophores with lower photoluminescence quantum yields demonstrated higher ECL. These findings are useful for further efforts in the search for more efficient ECL luminophores. PMID- 16270971 TI - Double-stranded metal-organic networks for one-dimensional mixed valence coordination polymers. AB - The design of new types of metal-organic networks and the search for unusual crystal architecture represents an important task for modern inorganic and materials chemistry research. A group of new monosubstituted phenylcyanoximes, containing F, Cl, and Br atoms at the 2, 3, or 4 positions, were synthesized using the high yield nitrosation reaction with CH3-ONO and were spectroscopically (1H NMR, 13C NMR, UV-visible, IR, mass spectrometry) and structurally characterized. Results of X-ray analysis revealed nonplanar trans-anti geometry for 2-chlorophenyl(oximino)acetonitrile, H(2Cl-PhCO); a nonplanar anti configuration for 4-chlorophenyl(oximino)acetonitrile, H(4Cl-PhCO); and planar cis-syn geometry for 3-fluorophenyl(oximino)acetonitrile, H(3F-PhCO). All arylcyanoximes undergo deprotonation in solutions with the formation of colored anions exhibiting pronounced negative solvatochromism in a series of polar protic and aprotic solvents. Nine thallium(I) cyanoximates were obtained using the reaction between hot (approximately 95 degrees C) aqueous solutions of Tl2CO3 and solid powdery monohalogenated arylcyanoximes HL. Crystal structures of two Tl(I) cyanoximates [Tl(2Cl-PhCO) and Tl(4Br-PhCO)] contained centrosymmetric dimeric units (TlL)2 that are connected to a coordination polymer by means of an oxygen atom of the oxime group of the neighboring molecule. Cyanoxime anions act as bridging ligands in both structures where the polymeric motif consists of double stranded Tl-O chains interconnected with the formation of zigzagging Tl2O2 planar rhombes. Thallium atoms form infinite linear arrays with close intermetallic separations. The nearest Tl(I)...Tl(I) distances are 3.838 and 4.058 angstroms in the Tl(2Cl-PhCO) and Tl(4Br-PhCO) structures, respectively, close to that in metallic thallium (3.456 angstroms). Monosubstituted phenyl groups are well aligned in pi-stacking columns that are perpendicular to the array of Tl(I) atoms and stabilize formed structures. Coordination polyhedrons of thallium(I) in these complexes represent distorted trigonal pyramids with stereoactive lone pair. PMID- 16270972 TI - Scanning probe microscopy characterization of single chains based on a one dimensional oxalato-bridged manganese(II) complex with 4-aminotriazole. AB - The compound [Mn(mu-ox)(4atr)2]n (1) (ox = oxalato and 4atr = 4-amine-1,2,4 triazole) has been synthesized and characterized by FT-IR spectroscopy, thermal analysis, variable-temperature magnetic measurements, and X-ray single-crystal diffraction methods. The crystal structure of compound 1 consists of one dimensional linear chains in which trans-[Mn(4atr)2]2+ units are sequentially bridged by centrosymmetric bis-bidentate oxalato ligands. Cryomagnetic measurements show an overall antiferromagnetic behavior of the compound. Isolated chains of this polymer have been obtained by sonication of 1 in ethanol or treatment of the polymer with NaOH and morphologically characterized on highly oriented pyrolitic graphite and mica surfaces by atomic force microscopy and scanning tunneling microscopy. The procedures employed to obtain single chains of this coordination polymer open a route for future nanotechnological applications of these types of materials. PMID- 16270973 TI - Sulfur K-edge XAS and DFT calculations on [Fe4S4]2+ clusters: effects of H bonding and structural distortion on covalency and spin topology. AB - Sulfur K-edge X-ray absorption spectroscopy of a hydrogen-bonded elongated [Fe4S4]2+ cube is reported. The data show that this synthetic cube is less covalent than a normal compressed cube with no hydrogen bonding. DFT calculations reveal that the observed difference in electronic structure has significant contributions from both the cluster distortion and from hydrogen bonding. The elongated and compressed Fe4S4 structures are found to have different spin topologies (i.e., orientation of the delocalized Fe2S2 subclusters which are antiferromagnetically coupled to each other). It is suggested that the H-bonding interaction with the counterion does not contribute to the cluster elongation. A magneto-structural correlation is developed for the Fe4S4 cube that is used to identify the redox-active Fe2S2 subclusters in active sites of HiPIP and ferredoxin proteins involving these clusters. PMID- 16270974 TI - Europium(III) and its halides in anhydrous room-temperature imidazolium-based ionic liquids: a combined TRES, EXAFS, and molecular dynamics study. AB - Combining spectroscopic techniques (TRES and EXAFS) and molecular dynamics simulations, we have investigated the state of trivalent europium dissolved in room-temperature ionic liquids (RTILs), as a function of the RTIL anion and in the presence of added chloride anions. The studied RTILs are based on the 1-butyl 3-methyl-imidazolium (Bumim+) cation and differ by their anionic counterparts: BF4-, PF6-, Tf- (triflate, CF3SO3-), and Tf2N- [(CF3SO2)2N-]. The results show the strong influence of the RTIL nature on the first solvation shell of europium and on its complexation with chloride. Depending on the RTIL, europium(III), which was introduced in solution as a triflate salt, is found to be solvated either by RTIL anions only or as neutral undissociated EuTf3 moieties completed by solvent anions. Kinetic effects, related to the viscosity of the RTIL and the nature of the europium salt, also markedly influence the coordination of added Cl or F- anions to the metal. PMID- 16270975 TI - Primary photoinduced processes in bimetallic dyads with extended aromatic bridges. tetraazatetrapyridopentacene complexes of ruthenium(II) and osmium(II). AB - The photophysics of the binuclear complexes [(phen)2M(tatpp)M(phen)2]4+, where M = Ru or Os, phen = 1,10-phenanthroline, and tatpp = 9,11,20,22 tetraazatetrapyrido[3,2-a:2'3'-c:3'',2''-l:2''',3''']pentacene, has been studied in acetonitrile and dichloromethane by femtosecond and nanosecond time-resolved techniques. The results demonstrate that complexes of different metals have different types of lowest excited state: a tatpp ligand-centered (LC) triplet in the case of Ru(II); a metal-to-ligand charge-transfer (MLCT) triplet state in the case of Os(II). The excited-state kinetics is strongly solvent-dependent. In the Ru(II) system, the formation and decay of the LC state take place, respectively, in 25 ps and ca. 5 ns in CH3CN and in 0.5 ps and 1.3 micros in CH2Cl2. These solvent effects can be rationalized on the basis of a thermally activated decay of the LC state through the upper MLCT state. In the Os(II) system, the formation and decay of the MLCT state take place, respectively, in 3.8 and 60 ps in CH3CN and in 0.5 and 4 ps in CH2Cl2. These effects are consistent with the solvent sensitivity of the MLCT energy, in terms of driving force and energy-gap law arguments. The relevance of these results for the use of ladder-type aromatic bridges as potential molecular wires is discussed. PMID- 16270976 TI - Synthesis of the five-coordinate ruthenium(II) complexes [(PCP)Ru(CO)(L)][BAr'4] [PCP = 2,6-(CH2P(t)Bu2)2C6H3, BAr'4 = 3,5-(CF3)2C6H3, L = eta1-ClCH2Cl, eta1-N2, or mu-Cl-Ru(PCP)(CO)]: reactions with phenyldiazomethane and phenylacetylene. AB - Reaction of (PCP)Ru(CO)(Cl) (1) with NaBAr'4 yields the bimetallic product [[(PCP)Ru(CO)](2)(mu-Cl)][BAr'4] (2). The monomeric five-coordinate complexes [(PCP)Ru(CO)(eta1-ClCH2Cl)][BAr'4] (3) and [(PCP)Ru(CO)(eta1-N2)][BAr'4] (4) are synthesized upon reaction of (PCP)Ru(CO)(OTf) (6) with NaBAr'4 in CH2Cl2 or C6H5F, respectively. The solid-state structures of 2, 3, and 4 have been determined by X-ray diffraction studies of single crystals. The reaction of 3 with PhCHN2 or PhCCH affords carbon-carbon coupling products involving the aryl group of the PCP ligand in transformations that likely proceed via the formation of Ru carbene or vinylidene intermediates. Density functional theory and hybrid quantum mechanics/molecular mechanics calculations were performed to investigate the bonding of weak bases to the 14-electron fragment [(PCP)Ru(CO)]+ and the energetics of different isomers of the product carbene and vinylidene complexes. PMID- 16270977 TI - Structural and chiroptical properties of the two coordination isomers of YbDOTA type complexes. AB - Studies of the structural, physical, and chemical properties of the lanthanide(III) complexes of DOTA (1,4,7,10-tetraazacyclododecane-1,4,7,10 tetraacetic acid) and related ligands are often complicated by the presence of two coordination isomers in solution. Since these coordination isomers are in exchange and cannot be separated, many techniques offer information only on the weighted average of the two isomers. Lanthanide ion complexes formed with the ligands S(RRRR)NO2BnDOTMA and S(SSSS)NO2BnDOTMA preferentially adopt only one of the two common coordination isomers in solution, so the ytterbium complexes of these ligands offer a unique opportunity to study the near-infrared circular dichroism (NIR-CD) characteristics of each coordination geometry in isolation. The spectra acquired support many of the conclusions and assumptions of previous studies and demonstrate that this spectroscopy is particularly sensitive to the distortion of the coordination polyhedron. This will have particularly relevant consequences when studying achiral YbDOTA-like systems forming labile adducts with (chiral) hosts and receptors. PMID- 16270978 TI - Hexacyanocobaltate(III) anions as precursors of Co(II)-Ni(II) cyano-bridged multidimensional assemblies: hydrothermal syntheses, crystal and powder X-ray structures, and magnetic properties. AB - Three novel cyanide-bridged heterobimetallic coordination polymers have been synthesized by hydrothermal routes, in superheated water solutions, by using K3[Co(CN)6], NiCl2.6H2O, and alpha-diimine ligands: [Ni(CN)4Co(phen)] (1; phen = 1,10-phenanthroline), [Ni(CN)4Co(2,2'-bipy)] (2; 2,2'-bipy = 2,2'-bipyiridine), and [Ni(CN)4Co(2,2'-bipy)2] (3). The isostructural compounds 1 and 2 contain a two-dimensional network with Co(II) centers octahedrally coordinated by one chelating 2,2'-bipy ligand and four cyanide groups of four distinct [Ni(CN)4]2-, through crystallographically equivalent, bridging units. Compound 3 contains one dimensional zigzag chains in which the Co(II) ion is coordinated by two chelating 2,2'-bipy ligands and two cyanides from two different [Ni(CN)4]2- units cis to each other. These compounds have been fully characterized by single-crystal or unconventional powder X-ray diffraction analyses and variable-temperature magnetic measurements. PMID- 16270979 TI - Revisiting the electronic structure of phosphazenes. AB - Natural bond orbital (NBO) and topological electron density analyses have been used to investigate the electronic structure of phosphazenes [N3P3R6] (R = H, F, Cl, Br, CH3, CF3, N(C2H4); 2R = O2C6H4), [N4P4Cl8], and H[NPCl2]4H. Using the former, the two most likely phosphazene bonding alternatives, negative hyperconjugation and ionic bonding have been critically evaluated. Ionic bonding, as suggested by topological analysis, was found to be the dominant bonding feature, although contributions from negative hyperconjugation are necessary for a more complete bonding description. Substituent effects on the P-N bond have been assessed and cases of bond length alternation have been rationalized using this combined bonding model, which supersedes previous models involving d-orbital participation, leading to an explanation for the observed bond length alternation found in some linear polyphosphazenes. In addition, common aromaticity indicators, nucleus independent chemical shifts (NICS) and para-delocalization indices (PDI), have been determined for the cyclophosphazenes. PMID- 16270980 TI - Characterization and comparison of Cm(III) and Eu(III) complexed with 2,6-di(5,6 dipropyl-1,2,4-triazin-3-yl)pyridine using EXAFS, TRFLS, and quantum-chemical methods. AB - The complexation of Cm(III) and Eu(III) with 2,6-di(5,6-dipropyl-1,2,4-triazin-3 yl)pyridine (n-C3H7-BTP) in nonaqueous organic solution is studied with extended X-ray absorption spectroscopy. Bond lengths are the same in both complexes. Quantum-chemical calculations performed at different levels support this finding. On the other hand, the Cm.(n-C3H7-BTP)3 complex is formed at much lower ligand-to metal concentration ratio than the Eu.(n-C3H7-BTP)3 complex, as shown by time resolved laser-induced fluorescence spectroscopy. This is in good agreement with n-C3H7-BTP's high selectivity for trivalent actinides over lanthanides in liquid liquid extraction. PMID- 16270981 TI - Computational study of sulfur atom-transfer reactions from thiiranes to ER3 (E = As, P; R = CH3, Ph). AB - Computational estimates have been made for the P=S and As=S bond strengths in triphenylphosphine sulfide and triphenylarsine sulfide, on the basis of G3 calculations for the methyl analogues and isodesmic-exchange reactions. Also, with the performance of the G3 method level for related compounds taken into consideration, the best estimates are 82 and 68 kcal/mol, respectively. While the value for triphenylarsine sulfide is within 2 kcal/mol of the single experimental estimate, that for triphenylphosphine sulfide is lower by 6 kcal/mol. (Capps, K. B.; Wixmerten, B.; Bauer, A.; Hoff, C. D. Inorg. Chem. 1998, 37, 2861-2864.) Despite virtually identical electronegativities of P and As, it is found that there is greater charge separation in the P=S bond. It is found that S atom transfer from thiiranes to arsines is exothermic. PMID- 16270982 TI - Synthesis, structure, and magnetic ordering of layered (2-D) V-based tris(oxalato)metalates. AB - The reaction of K3[M(III)(ox)3].3H2O [M = V (1), Cr; ox = oxalate], Mn(II)/V(II), and [N(n-Bu)4]Br in water leads to the isolation of 2-D V-based coordination polymers, [[N(n-Bu)4][Mn(II)V(III)(ox)3]]n (2), [[N(n-Bu)4][V(II)Cr(III)(ox)3]]n (3), [[N(n-Bu)4][V(II)V(III)(ox)3]]n (4), and an intermediate in the formation of 4, [[N(n-Bu)4][V(II)V(III)(ox)3(H2O)2]]n.2.5H2O (4a), while 1-D [V(II)(ox)(H2O)2]n (5) is obtained by using Na2ox and [V(OH2)6]SO4 in water. The structures of 1-5 have been investigated by single crystal and/or powder X-ray crystallography. In 1, V(III) is coordinated with three oxalate dianions as an approximately D3 symmetric, trigonally distorted octahedron. 1 is paramagnetic [mu(eff) = 2.68 mu(B) at 300 K, D = 3.84 cm(-1) (D/k(B) = 5.53 K), theta = -1.11 K, and g = 1.895], indicating an S = 1 ground state. 2 exhibits intralayer ferromagnetic coupling below 20 K, but does not magnetically order above 2 K, and 3 shows a strong antiferromagnetic interaction between V(II), S = 3/2 and Cr(III), S = 3/2 ions (theta = -116 K) within the 2-D layers. 4 and 4a magnetically order as ferrimagnets at T(c)'s, taken as the onset of magnetization, of 11 and 30 K, respectively. The 2 K remanent magnetizations are 2440 and 2230 emu.Oe mol(-1) and the coercive fields are 1460 and 4060 Oe for 4 and 4a, respectively. Both 4 and 4a clearly show frequency dependence, indicative of spin-glass-like behavior. The glass transition temperatures were at 6.3 and 27 K, respectively, for 4 and 4a. 1-D 5 exhibits antiferromagnetic coupling of -4.94 cm(-1) (H = -2Jsigma(i=1)n.S(i-1) - gmu(B)sigma(i=0)(n)H.S(i)) between the V(II) ions. PMID- 16270983 TI - Synthesis, structures, and DFT bonding analysis of new titanium hydrazido(2-) complexes. AB - The reaction of 1,1-diphenylhydrazine with Ti(NMe2)2Cl2 produced the monomeric terminal titanium hydrazido(2-) species Ti(NNPh2)Cl2(HNMe2)2 (1) in near quantitative yield. The reaction of Ti(NMe2)2Cl2 with the less sterically demanding ligand precursors 1,1-dimethylhydrazine or N-aminopiperidine gave the dimeric mu-eta2,eta1-bridged compounds Ti2(mu-eta2,eta1-NNMe2)2Cl4(HNMe2)2 (2) and Ti2[mu-eta2,eta1-NN(CH2)5]2Cl4(HNMe2)3 (3). The X-ray structures of 2 and 3 showed the formation of N-H...Cl hydrogen bonded dimers or chains, respectively. The reaction of 1 with an excess of pyridine formed [Ti(NNPh2)Cl2(py)2]n (4, n = 1 or 2). The reaction of the tert-butyl imido complex Ti(N(t)Bu)Cl2(py)3 with either 1,1-dimethylhydrazine or N-aminopiperidine again resulted in the formation of hydrazido-bridged dimeric complexes, namely Ti2(mu-eta2,eta1-NNMe2)2Cl4(py)2 (5, structurally characterized) and Ti2[mu-eta2,eta1-NN(CH2)5]2Cl4(py)2 (6). Compounds 1 and 4 are potential new entry points into terminal hydrazido(2-) chemistry of titanium. Compound 1 reacted with neutral fac-N3 donor ligands to form Ti(NNPh2)Cl2(Me3[9]aneN3) (7), Ti(NNPh2)Cl2(Me3[6]aneN3) (8), Ti(NNPh2)Cl2[HC(Me2pz)3] (9, structurally characterized), and Ti(NNPh2)Cl2[HC(n)Bupz)3] (10) in good yields (Me3[9]aneN3 = trimethyl-1,4,7 triazacyclononane, Me3[6]aneN3 = trimethyl-1,3,5-triazacyclohexane, HC(Me2pz)3 = tris(3,5-dimethylpyrazolyl)methane, and HC((n)Bupz)3 = tris(4 (n)butylpyrazolyl)methane). DFT calculations were performed on both the model terminal hydrazido compound Ti(NNPh2)Cl2[HC(pz)3] (I) and the corresponding imido compounds Ti(NMe)Cl2[HC(pz)3] (II) and Ti(NPh)Cl2[HC(pz)3] (III). The NNPh2 ligand binds to the metal center in an analogous manner to that of terminal imido ligands (metalligand triple bond), but with one of the Ti=N(alpha) pi components significantly destabilized by a pi interaction with the lone pair of the N(beta) atom. The NR ligand sigma donor ability was found to be NMe > NPh > NNPh2, whereas the overall (sigma + pi) donor ability is NMe > NNPh2 > NPh, as judged by fragment orbital populations, Ti-N atom-atom overlap populations, and fragment charge analysis. DFT calculations on the hydrazido ligand in a mu-eta2,eta1 bridging mode showed involvement of the N=N pi electrons in donation to one of the Ti centers. This TiN2 interaction is best represented as a metallocycle. PMID- 16270984 TI - Homoleptic trimethylsilylacetylide complexes of chromium(III), iron(II), and cobalt(III): syntheses, structures, and ligand field parameters. AB - A straightforward method for synthesizing soluble homoleptic trimethylsilylacetylide complexes of first-row transition metal ions is presented. Reaction of anhydrous CrCl2 with an excess of LiCCSiMe3 in THF at -25 degrees C affords orange Li3[Cr(CCSiMe3)6].6THF (1), while analogous reactions employing M(CF3SO3)2 (M = Fe or Co) generate pale yellow Li4[Fe(CCSiMe3)6].4LiCCSiMe3.4Et2O (2) and colorless Li3[Co(CCSiMe3)6].6THF (3). Slightly modified reaction conditions lead to Li8[Cr2O4(CCSiMe3)6].6LiCCSiMe3.4glyme (4), featuring a bis-mu-oxo-bridged binuclear complex, and Li3[Co(CCSiMe3)5(CCH)].LiCF3SO3.8THF (5). The crystal structures of 1-3 show the trimethylsilylacetylide complexes to display an octahedral coordination geometry, with M-C distances of 2.077(3), 1.917(7) 1.935(7), and 1.908(3) angstroms for M = Cr(III), Fe(II), and Co(III), respectively, and nearly linear M-C[triple bond]C angles. The UV-visible absorption spectrum of [Cr(CCSiMe3)6]3- in hexanes exhibits one spin-allowed d-d transition (4T2g <-- 4A1g) and three lower-energy spin-forbidden d-d transitions. The spectra of [Fe(CCSiMe3)6]4- and [Co(CCSiMe3)6]3- in acetonitrile display high intensity charge-transfer bands, which obscure all d-d transitions except for the lowest-energy spin-allowed band (1T1g <-- 1A1g) of the latter complex. Time dependent density functional theory (TD-DFT) calculations were employed as an aide in assigning the observed transitions. Taken together, the results are most consistent with the ligand field parameters delta(o) = 20,200 cm(-1) and B = 530 cm(-1) for [Cr(CCSiMe3)6]3-, delta(o) = 32 450 cm(-1) and B = 460 cm(-1) for [Fe(CCSiMe3)6]4- and delta(o) = 32 500 cm(-1) and B = 516 cm(-1) for [Co(CCSiMe3)6]3-. Ground-state DFT calculations support the conclusion that trimethylsilylacetylide acts as a pi-donor ligand. PMID- 16270986 TI - Multiconfigurational theoretical study of the octamethyldimetalates of Cr(II), Mo(II), W(II), and Re(III): revisiting the correlation between the M-M bond length and the delta --> delta* transition energy. AB - Four compounds containing metal-metal quadruple bonds, the [M2(CH3)8]n- ions (M = Cr, Mo, W, Re and n = 4, 4, 4, 2, respectively), have been studied theoretically using multiconfigurational quantum-chemical methods. The molecular structure of the ground state of these compounds has been determined and the energy of the delta --> delta* transition has been calculated and compared with previous experimental measurements. The high negative charges on the Cr, Mo, and W complexes lead to difficulties in the successful modeling of the ground-state structures, a problem that has been addressed by the explicit inclusion of four Li+ ions in these calculations. The ground-state geometries of the complexes and the delta --> delta* transition have been modeled with either excellent agreement with experiment (Re) or satisfactory agreement (Cr, Mo, and W). PMID- 16270987 TI - Structurally characterized ternary U-O-N compound, UN4O12: UO2(NO3)2.N2O4 or NO(+)UO2(NO3)3-? AB - The synthesis and characterization of the ternary U-O-N compound NO(+)UO2(NO3)3- (1) using IR and low-temperature and room-temperature Raman spectroscopy as well as 14N and 15N NMR spectroscopy are reported. In addition, solution Raman spectra of compound 1 recorded in various solvents are reported. The structure of compound 1 was determined using single-crystal X-ray diffraction techniques: monoclinic, C2/c, a = 13.3992(4) angstroms, b = 9.9781(4) angstroms, c = 7.6455(2) angstroms, beta = 115.452(2) degrees, V = 922.98(5) angstroms3, Z = 4. Compound 1 is highly moisture-sensitive and must be handled under an inert atmosphere. It reacts with water with the liberation of NO2. For the first time, this important precursor for the synthesis of anhydrous uranyl nitrate could be unambiguously identified and has been shown to be an ionic nitrosonium salt and not an adduct between uranyl nitrate and dinitrogen tetroxide, UO2(NO3)2.N2O4, as is incorrectly and predominantly cited in the literature. PMID- 16270985 TI - Reductive nitrosylation and proton-assisted bridge splitting of a (mu oxo)dimanganese(III) complex derived from a polypyridine ligand with one carboxamide group. AB - Aerobic oxidation of the Mn(II) complex [Mn(Papy3)(H2O)](ClO4) (1, PaPy3- is the anion of the designed ligand N,N-bis(2-pyridylmethyl)amine-N-ethyl-2-pyridine-2 carboxamide) in acetonitrile affords the (mu-oxo)dimanganese(III) complex [(Mn(PaPy3))2(mu-O)](ClO4)2 (3) in high yield. The unsupported single oxo bridge between the two high-spin Mn(III) centers in 3 is readily cleaved upon addition of proton sources such as phenol, acetic acid, and benzoic acid, and complexes of the type [Mn(PaPy3)(L)](ClO4) (5, L = PhO-; 6, L = AcO-; 7, L = BzO-) are formed. The basicity of the bridge is evident by the fact that simple addition of methanol to a solution of 3 in acetonitrile affords the methoxide complex [Mn(PaPy3)(OMe)](ClO4) (4). The structures of 3-5 and 7 have been determined. Passage of NO through a solution of 3 in acetonitrile produces the [Mn-NO]6 nitrosyl [Mn(PaPy3)(NO)](ClO4) (2) via reductive nitrosylation. Complexes 4-7 also afford the [Mn-NO]6 nitrosyl 2 upon reaction with NO. In the latter case, the anionic O-based ligands (such as MeO- and PhO-) act as built-in bases and promote reductive nitrosylation of the Mn(III) complexes. PMID- 16270988 TI - Local order in depolymerized silicate lattices. AB - In glycerol, near 200 degrees C, the silicate networks of mesoporous silicates and zeolites undergo a deep depolymerization process. In a few hours, depending on the initial concentration of the solid in glycerol and on the temperature, amorphous gels are obtained. In these gels, a fraction of the Si-O-Si bonds are transformed into Si-O-C. The constitutional aluminum remains bound to the silica network in the gel. The short range ordering is maintained to some extent: the size of the smallest structural unit in gels obtained from zeolites is in the range of the cubic nanometer, nm3. PMID- 16270990 TI - One-dimensional BiPO4 nanorods and two-dimensional BiOCl lamellae: fast low temperature sonochemical synthesis,characterization, and growth mechanism. AB - Regular BiPO4 nanorods, for the first time, and BiOCl lamellae have been successfully synthesized via a facile sonochemical method in a surfactant/ligand free system under ambient air. The as-prepared products are characterized by XRD, TEM, SAED, FE-SEM, HRTEM, and Raman spectroscopy. The effects of pH and ultrasound irradiation on the phase and morphology of the products are studied and the sonochemical formation mechanisms of 1D and 2D structures are discussed. TEM data from samples made after different reaction times suggest an ultrasound induced nucleation and an oriented-attachment growth mechanism. PMID- 16270989 TI - Factors controlling metal-ion selectivity in the binding sites of calcium-binding proteins. The metal-binding properties of amide donors. A crystallographic and thermodynamic study. AB - The metal-ion complexing properties of the ligand EDTAM (ethylenediamine N,N,N',N'-tetraacetamide) are investigated as a model for the role of amide oxygen donors in the binding sites of Ca-binding proteins. The structures of the complexes [Ca(EDTAM)NO3]NO3 (1), [La(EDTAM)(H2O)4](NO3)3.H2O (2), and [Cd(EDTAM)(NO3)]NO3 (3) are reported: 1 monoclinic, P2(1)/c, a = 10.853(2) angstroms, b = 12.893(3) angstroms, c = 13.407(3) angstroms, beta = 103.28(3) degrees, Z = 4, R = 0.0281; 2 triclinic, P, a = 8.695(2) angstroms, b = 9.960(2) angstroms, c = 16.136(3) angstroms, alpha = 95.57(3) degrees, beta = 94.84(3) degrees, gamma = 98.72(3) degrees, Z = 2, R = 0.0394; 3 monoclinic, P2(1)/c, a = 10.767(2) angstroms, b = 12.952(2) angstroms, c = 13.273(2) angstroms, beta = 103.572(3) degrees, Z = 4, R = 0.0167. Compounds 1 and 3 are isostructural, and the EDTAM binds to the metal ion through its two N-donors and four O-donors from the amide groups. Ca(II) in 1 is 8-coordinate with a chelating NO3- group, while Cd(II) in 3 may possibly be 7-coordinate, with an asymmetrically coordinated NO3- that is best regarded as unidentate. The La(III) in 2 is coordinated to the EDTAM in a manner similar to that of 1 and 3, but it is 10-coordinate with four water molecules coordinated to the La(III). The formation constants (log K1) for complexes of a variety of metal ions with EDTAM are reported in 0.1 M NaNO3 at 25.0 +/- 0.1 degrees C. These are compared to the log K1 values for en (ethylenediamine) and THPED (N,N,N',N'-tetrakis(2-hydroxypropyl) ethylenediamine). For large metal ions, such as Ca2+ or La3+, log K1 increases strongly when the four acetamide groups are added to en to give EDTAM, whereas for a small metal ion, such as Mg2+, this increase is small. The log K1 values for EDTAM compared to THPED suggest that the amide oxygen is a much stronger base than the alcoholic oxygen. Structures of binding sites in 40 Ca-binding proteins are examined. It is shown that the Ca-O=C bond angles involving coordinated amides in these sites are large, commonly being in the 150-180 degrees range. This is discussed in terms of the idea that for purely ionic bonding the M-O=C bond angle will approach 180 degrees, while for covalent bonding the angle should be closer to 120 degrees. How this fact might be used by the proteins to control selectivity for different metal ions is discussed. PMID- 16270991 TI - Disilver(I) rectangular-shaped metallacycles: X-ray crystal structure and dynamic behavior in solution. AB - Reaction of the ditopic semirigid ligand 1,2-bis(imidazolylmethyl)benzene (1,2 bImb) or the flexible ligand 1,4-bis(2-benzimidazolyl)butane (C4BIm) with AgX (X = ClO4-, BF4-, CF3CO2-) afforded five new complexes, namely, [Ag2(1,2 bImb)2](ClO4)2 (1), [Ag2(1,2-bImb)2](BF4)2 (2), [Ag2(1,2-bImb)2](CF3CO2)2.2CH3OH (3.2CH3OH), [Ag2(C4BIm)2](ClO4)2.2DMF (4.2DMF), and [Ag2(C4BIm)2](CF3CO2)2.2H2O (5.2H2O), all of which contain a centrosymmetric, rectangular-shaped cationic disilver(I) metallacycle [Ag2(L)2]2+. In 1-3, a pair of 1,2-bImb ligands takes on the syn conformation to connect two Ag(I) ions to give a compressed rectangle with a transannular Ag...Ag separation of 3.27-3.36 angstroms, whereas in 4 and 5, the pair of planar C4BIm ligands acts in the cis conformation to connect two Ag(I) ions to yield a normal rectangle with a transannular Ag...Ag separation of 7.67-7.91 angstroms. The anions form Ag...O or Ag...F weak interactions in 1-3 and O-H...O or N-H...O hydrogen bonds in 4 and 5 in crystal packing but exhibit no significant influence on the formation of the disilver(I) macrocycles. The solution structure and dynamic behavior of the complexes studied by electrospray ionization mass spectrometry, 1H NMR, and variable-temperature NMR indicated that the dynamic equilibrium between the [Ag2(L)2]2+ cation and the open-ring oligomers or other potential species occurs via solvent-assisted dissociative exchange. The metal-ligand exchange barrier was estimated to be 54.5 kJ mol(-1). PMID- 16270992 TI - Autoxidation-product-initiated dioxygenases: vanadium-based, record catalytic lifetime catechol dioxygenase catalysis. AB - In recent work, it was shown that V-containing polyoxometalates such as (n Bu4N)7SiW9V3O40 or (n-Bu4N)9P2W15V3O62, as well as eight other V-containing precatalysts tested, evolve to a high activity, long catalytic lifetime (> or = 30,000-100,000 total turnovers) 3,5-di-tert-butylcatechol dioxygenase, in which Pierpont's complex [VO(DBSQ)(DTBC)]2 (where DBSQ is 3,5-di-tert-butylsemiquinone and DTBC is the 3,5-di-tert-butylcatecholate dianion) was identified as a common catalyst or catalyst resting state (Yin, C.-X.; Finke, R. G. Vanadium-Based, Extended Catalytic Lifetime Catechol Dioxygenases: Evidence For a Common Catalyst. J. Am. Chem. Soc. 2005, 127 (25), 9003-9013). Herein, those findings are followed up by studies aimed at answering the following questions about this record catalytic lifetime 3,5-di-tert-butylcatechol dioxygenase catalyst: (i) What is the key to how V leaches from, for example, seemingly robust V-containing polyoxometalate precatalysts? (ii) What is the key to the sigmoidal, apparently autocatalytic kinetics observed? (iii) What can be learned about the underlying reactions that form [VO(DBSQ)(DTBC)]2? (iv) Finally, do the answers to (i-iii) lead to any broader insights or concepts? Key findings from the present work include the fact that the reaction involves a novel, autoxidation-product-induced dioxygenase, that is, one in which the undesired autoxidation of the 3,5-di-tert butylcatechol substrate to the corresponding benzoquinone and H2O2 turns on the desired dioxygenase catalysis via a V-leaching process which eventually yields Pierpont's complex, [VO(DBSQ)(DTBC)]2. Plausible reactions en route to [VO(DBSQ)(DTBC)]2 consistent with the kinetic data, the role of H2O2, and the relevant literature are provided. The results provide a prototype example of the little observed but likely more general concept of an autoxidation-product initiated reaction. The results also provide considerable simplification of, and insight into, the previously disparate literature of V-based 3,5-di-tert butylcatechol dioxygenase catalysis. PMID- 16270993 TI - Tris(thioimidazolyl)borate-zinc-thiolate complexes for the modeling of biological thiolate alkylations. AB - The S3Zn-SR coordination of thiolate-alkylating enzymes such as the Ada DNA repair protein was reproduced in tris(thioimidazolyl)borate-zinc-thiolate complexes Tti(R)Zn-SR'. Four different Tti(R) ligands and nine different thiolates were employed, yielding a total of 12 new complexes. In addition, one Tti(R)Zn-SH complex and two thiolate-bridged [Tti(R)-SEt-Tti(R)]+ complexes were obtained. A selection of six thiolate complexes was converted with methyl iodide to the corresponding methyl thioethers and Tti(R)Zn-I. According to a kinetic analysis these reactions are second-order processes, which implies that the alkylations are likely to occur at the zinc-bound thiolates. They are much faster than the alkylations of zinc thiolates with N3 or N2S tripod ligands. The most reactive thiolate, Tti(Xyl)Zn-SEt, reacts slowly with trimethyl phosphate in a nonpolar medium at room temperature, yielding methyl-ethyl-thioether and Tti(Xyl)Zn-OPO(OMe)2 which can be converted back to the thiolate complex with NaSEt. This is the closest reproduction of the Ada repair process so far. PMID- 16270994 TI - Tetrakis(thiadiazole)porphyrazines. 4. Direct template synthesis, structure, general physicochemical behavior, and redox properties of Al(III), Ga(III), and In(III) complexes. AB - Monometallic derivatives of tetrakis(1,2,5-thiadiazole)porphyrazine, [TTDPzH2], with main group tervalent metal ions having the formulae [TTDPzMX] (TTDPz = tetrakis(1,2,5-thiadiazole)porphyrazinato dianion; M = Al(III), X = Cl-, Br-, OH ; M = Ga(III), X = Cl-, OH-; M = In(III), X = AcO-) were prepared and investigated by single-crystal X-ray analysis and IR and UV-vis spectroscopy as well as cyclic voltammetry and spectroelectrochemistry. The complexes [TTDPzMX] (M = Al(III), X = Cl-, Br-; M = Ga(III), X = Cl-) were obtained by direct autocyclotetramerization of the precursor 3,4-dicyano-1,2,5-thiadiazole in hot quinoline in the presence of MX3 salts (M = Al(III), Ga(III); X = Cl-, Br-) and were hydrolized to form the corresponding hydroxide derivatives, [TTDPzMOH]. The In(III) complex, [TTDPzIn(OAc)], was obtained from the free-base macrocycle [TTDPzH2] with In(OH)(OAc)2 in CH3COOH. A single-crystal X-ray study was made at 173 K on the two isostructural species [TTDPzMCl] (M = Al(III), Ga(III)), which have space group P, with a = 12.470(14), b = 12.464(13), and c = 13.947(12) angstroms, alpha = 70.72(3), beta = 79.76(3), and gamma = 90.06(3) degrees, V = 2009.3(3) angstroms3, and Z = 4 for [TTDPzAlCl] and a = 12.429(3), b = 12.430(3), and c = 13.851(3) angstroms, alpha = 70.663(6), beta = 79.788(8), and gamma = 89.991(9) degrees, V = 1983.3(7) angstroms3, and Z = 4 for [TTDPzGaCl]. Square pyramidal coordination exists about the M(III) centers, with Cl- occupying the apical position (Al-Cl = 2.171(5) and Ga-Cl = 2.193(1) angstroms). Al(III) and Ga(III) are located at distances of 0.416(6) and 0.444(2) angstroms from the center of the N4 system. The molecular packing consists of stacked double layers with internal and external average interlayer distances of 3.2 and 3.3 angstroms, respectively. IR spectra show nu(Al-Cl) at 345 cm(-1) for [TTDPzAlCl], nu(Al-Br) at 330 cm(-1) for [TTDPzAlBr], and nu(Ga-Cl) at 382 cm(-1) for [TTDPzGaCl]. The UV-vis spectra in weakly basic (pyridine, DMF, DMSO) and acidic solvents (CF3COOH, H2SO4) show the typical intense pi --> pi transition bands in the Soret (300-400 nm) and Q-band regions (640-660 nm), the bands evidencing some dependence on the nature of the solvent, particularly in acidic solutions. Cyclic voltammetry, differential pulse voltammetry, and thin-layer spectroelectrochemical measurements in pyridine and dimethylformamide of the species [TTDPzMX] indicate reversible first and second one-electron reductions, whereas additional ill-defined reductions are observed at more negative potentials. The examined species are much easier to reduce than their phthalocyanine or porphyrin analogues as a result of the remarkable electron attracting properties of the TTDPz macrocycle which contains annulated strongly electron-deficient thiadiazole rings. PMID- 16270995 TI - Synthesis of the pivalamidate-bridged pentanuclear platinum(II,III) linear complexes with Pt...Pt interactions. AB - Pentanuclear linear chain Pt(II,III) complexes [[Pt2(NH3)2X2((CH3)3CCONH)2(CH2COCH3)]2[PtX'4]].nCH3COCH3 (X = X' = Cl, n = 2 (1a), X = Cl, X' = Br, n = 1 (1b), X = Br, X' = Cl, n = 2 (1c), X = X' = Br, n = 1 (1d)) composed of a monomeric Pt(II) complex sandwiched by two amidate-bridged Pt dimers were synthesized from the reaction of the acetonyl dinuclear Pt(III) complexes having equatorial halide ligands [Pt2(NH3)2X2((CH3)3CCONH)2(CH2COCH3)]X' ' (X = Cl (2a), Br (2b), X' ' = NO3-, CH3C6H4SO3-, BF4-, PF6-, ClO4-), with K2[PtX'4] (X' = Cl, Br). The X-ray structures of 1a-1d show that the complexes have metal-metal bonded linear Pt5 structures, and the oxidation state of the metals is approximately Pt(III) Pt(III)...Pt(II)...Pt(III)-Pt(III). The Pt...Pt interactions between the dimer units and the monomer are due to the induced Pt(II)-Pt(IV) polarization of the Pt(III) dimeric unit caused by the electron withdrawal of the equatorial halide ligands. The density functional theory calculation clearly shows that the Pt...Pt interactions between the dimers and the monomer are made by the electron transfer from the monomer to the dimers. The pentanuclear complexes have flexible Pt backbones with the Pt chain adopting either arch or sigmoid structures depending on the crystal packing. PMID- 16270996 TI - Synthesis and structure of a distorted octahedral palladium(II) complex coordinated with a tetrathioether ligand tethered with bulky substituents. AB - A new type of an o-phenylene-bridged tetrathioether ligand tethered with extremely bulky substituents, 2,4,6-tris[bis(trimethylsilyl)methyl]phenyl (Tbt) groups, at its terminal sulfur atoms, TbtS[(o-phenylene)S]3Tbt (1), was synthesized by taking advantage of the coupling reaction of thiols with iodobenzenes using Cu2O in 2,4,6-trimethylpyridine. Complexation of 1 with Na2PdCl4 gave the corresponding dichloropalladium(II) complex, [PdCl2(1)] (7). The X-ray structural analysis of 7 indicated that the central palladium metal is in a distorted octahedral environment, where the two inner sulfur atoms of 1 and the two chlorine atoms form a square planar arrangement around the palladium metal and the two terminal sulfur atoms of 1 weakly coordinate to the palladium center at the axial positions. In addition, a phenyl analogue of 1, PhS[(o phenylene)S]3Ph (2), was synthesized by a method similar to that for 1. Reaction of 2 with Na2PdCl4 gave the corresponding dichloropalladium(II) complex, [PdCl2(2)] (8). X-ray crystallography of 8 showed a type of the structure different from the distorted octahedral structure in 7, i.e., a square planar arrangement around the central palladium atom with the one terminal sulfur atom of 2, its neighboring sulfur atom, and the two chlorine atoms. The results of the NMR studies on 8 in a CDCl3 solution were not consistent with the results of the X-ray crystallography and suggested the coordination of the two inner sulfur atoms of 2 to the palladium metal, although a possibility of the existence of the rapid interconversion among isomers could not be excluded. PMID- 16270997 TI - 1,2-Dicarba-closo-dodecaboran-1-yl naphthalene derivatives. AB - 1,2-Dicarba-closo-dodecaboranes (o-carboranes) and naphthalenes have potential value as components or building blocks for supramolecular systems. We have efficiently synthesized 1-(1,2-dicarba-closo-dodecaboran-1-yl)naphthalene and 2 (1,2-dicarba-closo-dodecaboran-1-yl)naphthalene derivatives by employing three preparative methods: cyclization of the corresponding acetylenes with decaborane(14), an Ullmann-type coupling reaction of carboranes with aryl halide, and the aromatic nucleophilic substitution (S(N)Ar) reaction of aryl-o-carboranes with nitrophenyl halide. The optimum conditions of each method for synthesis of the title compounds were also investigated. PMID- 16270999 TI - Efficient photoreduction process of [Ru3(mu3-O)(mu-CH3CO2)6L3]+ by electron mediation via the viologen dication by excitation of a zinc porphyrin. AB - Photoinduced electron-transfer and electron-mediation processes from the excited triplet state of zinc tetraphenylporphyrin (3ZnTPP) to the hexyl viologen dication (HV2+) in the presence of oxo-acetato-bridged triruthenium clusters, [Ru3(mu3-O)(mu-CH3CO2)6L3]+, have been revealed by the transient absorption spectra in the visible and near-IR regions. By the nanosecond laser-flash photolysis of ZnTPP in the presence of HV2+ and [Ru3(mu3-O)(mu-CH3CO2)6L3]+, the transient absorption bands of the radical cation of ZnTPP (ZnTPP*+) and the reduced viologen (HV*+) were initially observed with the concomitant decay of 3ZnTPP, after which an extra electron of HV*+ mediates to [Ru3(mu3-O)(mu CH3CO2)6L3]+, efficiently generating [Ru3(mu3-O)(mu-CH3CO2)6L3]0 with high potential. Although back-electron transfer took place between ZnTPP*+ and [Ru3(mu3-O)(mu-CH3CO2)6L3]0 in the diffusion-controlled limit, [Ru3(mu3-O)(mu CH3CO2)6L3]0 accumulates at a steady concentration upon further addition of 1 benzyl-1,4-dihydronicotinamide (BNAH) as a sacrificial donor to re-produce ZnTPP from ZnTPP*+. Therefore, we established a novel system to accumulate [Ru3(mu3 O)(mu-CH3CO2)6L3]0 as an electron pool by the excitation of ZnTPP as photosensitizing electron donor in the presence of HV2+ and BNAH as an electron mediating reagent and sacrificial donor, respectively. With the increase in the electron-withdrawing abilities of the ligands, the final yields of [Ru3(mu3-O)(mu CH3CO2)6L3]0 increased. PMID- 16270998 TI - Spectroscopic and thermodynamic characterization of the E151D and E151A altered leucine aminopeptidases from Aeromonas proteolytica. AB - Previous kinetic characterization of the glutamate 151 (E151)-substituted forms of the leucine aminopeptidase from Aeromonas proteolytica (Vibrio proteolyticus; AAP) has provided critical evidence that this residue functions as the general acid/base. The close proximity of similar glutamate residues to the bridging water/hydroxide of the dinuclear active sites of metalloenzymes (2.80 and 3.94 angstroms in carboxypeptidase G2 and 3.30 and 3.63 angstroms in AAP), suggests it may also be involved in stabilizing the active-site metal ions. Therefore, the structural perturbations of the dinuclear active site of AAP were examined for two E151-substituted forms, namely E151D-AAP and E151A-AAP, by UV-vis and electron paramagnetic resonance (EPR) spectroscopy. UV-vis spectroscopy of Co(II) substituted E151A-AAP did not reveal any significant changes in the electronic absorption spectra. However UV-vis spectra of mono- and dicobalt(II) E151D-AAP exhibited a lower molecular absorptivity compared to AAP (23 and 43 M(-1) cm(-1) vs. 56 and 109 M(-1) cm(-1) for E151D-AAP and AAP, respectively) suggesting both Co(II) ions reside in distorted octahedral coordination geometry in E151D-AAP. EPR spectra of [Co_(E151D-AAP)], [ZnCo(E151D-AAP)], and [(CoCo(E151D-AAP)] were identical, with g(perpendicular) = 2.35, g(parallel) = 2.19, and E/D = 0.19, similar to [CoCo(AAP)]. On the other hand, the EPR spectrum of [Co_(E151A-AAP)] was best simulated assuming the presence of two species with (i) g(x,y) = 2.509, g(z) = 2.19, E/D = 0.19, A = 0.0069 cm(-1) and (ii) g(x,y) = 2.565, g(z) = 2.19, E/D = 0.20, A = 0.0082 cm(-1) indicative of a five- or six-coordinate species. Isothermal titration calorimetry experiments revealed a large decrease in Zn(II) affinities, with K(d) values elevated by factors of approximately 850 and approximately 24,000 for the first metal binding events of E151D- and E151A-AAP, respectively. The combination of these data indicates that E151 serves to stabilize the dinuclear active site of AAP. PMID- 16271000 TI - Heme axial methionine fluxion in Pseudomonas aeruginosa Asn64Gln cytochrome c551. AB - Heme axial methionine ligands in ferricytochromes c552 from Hydrogenobacter thermophilus (HT) and Nitrosomonas europaea, both members of the cyt c8 family, display fluxional behavior. The ligand motion, proposed to be inversion at sulfur, results in an unusually small range of hyperfine shifts for heme substituents in these proteins. Herein, heme axial Met fluxion is induced in a structurally homologous cytochrome c551 from Pseudomonas aeruginosa (PA) by substituting heme pocket residue Asn64 with Gln. The mutant, PA-N64Q, displays a highly compressed range of heme substituent hyperfine shifts, temperature dependent heme methyl resonance line broadening, low rhombic magnetic anisotropy, and a magnetic axes orientation consistent with Met orientational averaging. Analysis of NMR properties of PA-N64Q demonstrates that the heme pocket of the mutant resembles that of HT. This result confirms the importance of peripheral interactions and, in particular, residue 64 in determining axial Met orientation and heme electronic structure in proteins in the cyt c8 family. PMID- 16271001 TI - Oxidative addition of methyl iodide to a new type of binuclear platinum(II) complex: a kinetic study. AB - A new organodiplatinum(II) complex cis,cis-[Me2Pt(mu-NN)(mu-dppm)PtMe2] (1), in which NN = phthalazine and dppm = bis(diphenylphosphino)methane, is synthesized by the reaction of cis,cis-[Me2Pt(mu-SMe2)(mu-dppm)PtMe2] with 1 equiv of NN. Complex 1 has a 5d(pi)(Pt) --> pi(imine) metal-to-ligand charge-transfer band in the visible region, which was used to easily follow the kinetics of its reaction with MeI. Meanwhile, the complex contains a robust bridging dppm ligand that holds the binuclear integrity during the reaction. A double MeI oxidative addition was observed, as shown by spectrophotometry and confirmed by a low temperature 31P NMR study. The classical S(N)2 mechanism was suggested for both steps, and the involved intermediates were suggested. Consistent with the proposed mechanism, the rates of the reactions at different temperatures were slower in benzene than in acetone and large negative deltaS values were found in each step. However, some abnormalities were observed in the related rate constants and deltaS values, which were demonstrated to be due to the associative involvement of the polar acetone molecules in the reactions. The rates are almost 6 times slower in the second step as compared to the first step because of the electronic effects transmitted through the ligands and the steric effects. PMID- 16271002 TI - Wurster's thiacrown ethers: synthesis, properties, and Pt(II)-coordination chemistry. AB - Two isomeric redox-responsive azathiacrown ethers, based on p-phenylenediamine, have been synthesized in traditional crown (L1) and crownophane (L2) architectures. Each of these "Wurster's crowns" was designed to target the encapsulation of transition or heavy metal ions. The solid-state structures of these ligands show binding cavities defined by three exocyclic sulfur atoms and either a N donor atom (L1) or the electron-rich pi face of the phenylenediamine subunit (L2). The ability of these ligands to form complexes with platinum(II) was investigated by various techniques including 1H NMR spectroscopy, electrospray mass spectrometry, cyclic voltammetry, and single-crystal X-ray analysis. The traditional crown geometry proved to be better at forming stable endocyclic complexes with Pt(II) than the crownophane geometry. The square-planar Pt(II) crown complex includes direct bonding to the redox center (Pt1-N1 = 2.125 angstroms and Pt1-S(av) = 2.278 angstroms) with concomitant polarization of the phenylenediamine moiety. This results in the crown complex oxidizing 916 mV more anodically than the free ligand. In contrast, modest shifts in the oxidation potential of the crownophane isomer indicate poor interaction between the redox center and complexed Pt(II) ion. PMID- 16271003 TI - Formation and characterization of thorium methylidene CH2=ThHX complexes. AB - Laser-ablated thorium atoms react with methyl fluoride to give the CH2=ThHF molecule as the major product observed and trapped in solid argon. Infrared spectroscopy, isotopic substitution, and density functional theoretical frequency calculations confirm the identification of this methylidene complex. The four strongest computed absorptions (Th-H stretch, Th=C stretch, CH2 wag, and Th-F stretch) are the four vibrational modes observed. The CH2=ThHCl and CH2=ThHBr species formed from methyl chloride and methyl bromide exhibit the first three of these modes in the infrared spectra. The computed structures (B3LYP and CCSD) show considerable agostic interaction, similar to that observed for the Group 4 CH2=MHX (M = Ti, Zr, Hf) methylidene complexes, and the agostic angle and C=Th bond length decrease slightly in the CH2=ThHX series (X = F, Cl, Br). PMID- 16271004 TI - Synthesis, structural characterization, and computational studies of novel diiodine adducts with the heterocyclic thioamides N-methylbenzothiazole-2-thione and benzimidazole-2-thione: implications with the mechanism of action of antithyroid drugs. AB - Reaction of N-methylbenzothiazole-2-thione (C8H7NS2 or NMBZT) with diiodine produced the charge-transfer (ct) complex [(NMBZT).I2] (1). NMBZT reacts with diiodine in the presence of FeCl3 in a molar ratio of 3:6:1 and forms the ionic complex [[(NMBZT)2I+].[FeCl4]-] (2) together with [[(NMBZT)2I+].[I7]-] (2a) iodonium salt. The reaction of benzimidazole-2-thione (C7H6N2S or MBZIM) with diiodine on the other hand results in the formation of the ct [[(MBZIM)2I]+[I3] ].[(MBZIM).I2] (3) compound. The compounds have been characterized by elemental analyses, DTA-TG, FT-Raman, FT-IR, UV-vis, and 1H NMR spectroscopies, and X-ray crystal structure determinations. Compound 1, C8H7I2NS2, is orthorhombic with a space group Pna2(1) and a = 12.5147(13) angstroms, b = 22.536(3) angstroms, c = 4.2994(5) angstroms, and Z = 4. Compound 2, C16H14Cl4FeIN2S4, is monoclinic, space group C2/c, a = 35.781(2) angstroms, b = 7.4761(5) angstroms, c = 18.4677(12) angstroms, beta = 107.219(1) degrees, and Z = 8. Compound 3, C21H18I6N6S3, monoclinic, space group P2(1)/n, a = 14.0652(11) angstroms, b = 22.536(3) angstroms, c = 4.2994(5) angstroms, beta = 99.635(7) degrees, and Z = 4, consists of two component moieties cocrystallized, one neutral which contains the benzimidazole-2-thione (MBIZM) ligand bonded with an iodine atom through sulfur, forming a compound with a "spoke" structure [(MBZIM)I2] 3a, while the other is the ionic complex [[(MBZII)2I+].[I3]-] (3b). The X-ray crystal structure of 1 shows a bond between the thione-sulfur atom and one of the iodine atoms in an essentially planar arrangement. In the cation of 2, an iodine is coordinated by two thione-sulfur atoms in a linear arrangement but the molecule is not planar. For the first time in the solid state a spoke-ionic mixed complex has been characterized in 3. One component of the structure is a molecular diiodine adduct, i.e., [(MBZIM)I2] (3a), with a linear coordination geometry in a decidedly planar arrangement, and the other component is an ionic adduct [[(MBZIM)2I]+.[I3]-] (3b) with the cation having an arrangement similar to that found for 1. Theoretical calculations using density functional (DFT) and ab initio Hartree-Fock theory have been carried out for 1 and 3a,b. The results are consistent with the experimental data. Conclusions on the behavior of a thioamide, when used as an antithyroid drug, have also been made. PMID- 16271005 TI - Biferrocene-M(mnt)2 charge-transfer complexes (M = Ni, Co; mnt = maleonitriledithiolate). structure, valence states, and magnetic properties. AB - Charge-transfer salts of branched-alkyl biferrocenes, (1',1' ''-R2-1,1' ' biferrocene)[Ni(mnt)2] (1a, R = isopropyl; 2a, R = dineopentyl) and (1',1' ''-R2 1,1' '-biferrocene)2[Co(mnt)2]2 (1b, R = isopropyl; 2b, R = dineopentyl), were prepared. Their valence states were investigated using X-ray crystallography and Mossbauer spectroscopy. Complexes 1a and 1b show segregated-stack crystal structures that contain columns of acceptors, whereas structures of 2a and 2b, which contain bulky donors, are rather discrete. All of the complexes contain mixed-valent biferrocenium monocations. A two-step valence transition was found in complex 1a. The crystal contains two crystallographically independent cations: one undergoes valence localization below room temperature; the other undergoes valence localization below ca. 130 K. The former transition is derived from asymmetry of the crystal environment around the cation, whereas the latter one is caused by symmetry lowering coupled with a spin-Peierls transition (T(C) = 133.2 K) associated with the dimerization of the acceptors. This compound was found to exhibit a dielectric response based on valence tautomerization. Other complexes (1b, 2a, and 2b) show a valence-trapped state. In all complexes, charge localization was found to occur through local electrostatic interactions between the donor's cationic moiety and the acceptor's electronegative moieties. PMID- 16271007 TI - Cognitive determinants of energy balance-related behaviours: measurement issues. AB - The burden of disease as a result of overweight and obesity calls for in-depth examination of the main causes of behavioural actions responsible for weight gain. Since weight gain is the result of a positive energy balance, these behavioural actions are referred to as 'energy balance-related behaviours' (EBRBs). In the broadest sense, there are only two EBRBs: food intake and physical activity. However, both diet and physical activity are complex behavioural categories that involve a variety of actions. This article discusses the potential problems and opportunities related to the assessment of cognitive determinants of energy intake and energy expenditure behaviours. We argue for the necessity of studying determinants of EBRBs within an energy balance approach, i.e. focusing on energy input as well as output, instead of only studying dietary change or physical activity behaviour. As a result, however, theoretically sound questionnaires assessing determinants of EBRBs are likely to annoy respondents. It is especially the measurement of the behaviours and the use of belief-based constructs that cause questionnaires to be long, which may lead to low response rates and invalid data. In this article, we propose a careful and systematic consideration of the inclusion or exclusion of measures of cognitive determinants. First, if studies show that an EBRB is strongly influenced by environmental factors and is not or only to a minor extent under intentional control, measurement of cognitions is of little use. Second, only when we have proof that attitudes, norms and perceived behavioural control predict intentions, should we aim to assess the underlying beliefs. Third, since assessment of beliefs results in similar or better prediction than using belief-valuation combinations, we should not 'annoy' respondents with valuation items. Finally, we argue that the traditional paper-and-pencil survey is still the most reliable and practical data collection method. However, pilot studies applying computerised adaptive methods to determinants of EBRBs are encouraged. PMID- 16271008 TI - A nonlinear dynamic approach for evaluating postural control: new directions for the management of sport-related cerebral concussion. AB - Recent research suggests that traditional biomechanical models of postural stability do not fully characterise the nonlinear properties of postural control. In sports medicine, this limitation is manifest in the postural steadiness assessment approach, which may not be sufficient for detecting the presence of subtle physiological change after injury. The limitation is especially relevant given that return-to-play decisions are being made based on assessment results. This update first reviews the theoretical foundation and limitations of the traditional postural stability paradigm. It then offers, using the clinical example of athletes recovering from cerebral concussion, an alternative theoretical proposition for measuring changes in postural control by applying a nonlinear dynamic measure known as 'approximate entropy'. Approximate entropy shows promise as a valuable means of detecting previously unrecognised, subtle physiological changes after concussion. It is recommended as an important supplemental assessment tool for determining an athlete's readiness to resume competitive activity. PMID- 16271010 TI - Possible stimuli for strength and power adaptation: acute mechanical responses. AB - A great deal of literature has investigated the effects of various resistance training programmes on strength and power changes. Surprisingly, however, our understanding of the stimuli that affect adaptation still remains relatively unexplained. It is thought that strength and power adaptation is mediated by mechanical stimuli, that is the kinematics and kinetics associated with resistance exercise (e.g. forces, contraction duration, power and work), and their interaction with other hormonal and metabolic factors. However, the effect of different combinations of kinematic and kinetic variables and their contribution to adaptation is unclear. The mechanical response to single repetitions has been investigated by a number of researchers; however, it seems problematic to extrapolate the findings of this type of research to the responses associated with a typical resistance training session. That is, resistance training is typified by multiple repetitions, sets and exercises, rest periods of varying durations and different movement techniques (e.g. controlled and explosive). Understanding the mechanical stimuli afforded by such loading schemes would intuitively lead to a better appreciation of how various mechanical stimuli affect adaptation. It will be evident throughout this article that very little research has adopted such an approach; hence our understanding in this area remains rudimentary at best. One should therefore remain cognizant of the limitations that exist in the interpretation of research in this field. We contend that strength and power research needs to adopt a set kinematic and kinetic analysis to improve our understanding of how to optimise strength and power. PMID- 16271009 TI - Electrical stimulation superimposed onto voluntary muscular contraction. AB - Electrical stimulation (ES) reverses the order of recruitment of motor units (MU) observed with voluntary muscular contraction (VOL) since under ES, large MU are recruited before small MU. The superimposition of ES onto VOL (superimposed technique: application of an electrical stimulus during a voluntary muscle action) can theoretically activate more motor units than VOL performed alone, which can engender an increase of the contraction force. Two superimposed techniques can be used: (i) the twitch interpolation technique (ITT), which consists of interjecting an electrical stimulus onto the muscle nerve; and (ii) the percutaneous superimposed electrical stimulation technique (PST), where the stimulation is applied to the muscle belly. These two superimposed techniques can be used to evaluate the ability to fully activate a muscle. They can thus be employed to distinguish the central or peripheral nature of fatigue after exhausting exercise. In general, whatever the technique employed, the superimposition of ES onto volitional exercise does not recruit more MU than VOL, except with eccentric actions. Nevertheless, the neuromuscular response associated with the use of the superimposed technique (ITT and PST) depends on the parameter of the superimposed current. The sex and the training level of the subjects can also modify the physiological impact of the superimposed technique. Although the motor control differs drastically between training with ES and VOL, the integration of the superimposed technique in training programmes with healthy subjects does not reveal significant benefits compared with programmes performed only with voluntary exercises. Nevertheless, in a therapeutic context, training programmes using ES superimposition compensate volume and muscle strength deficit with more efficiency than programmes using VOL or ES separately. PMID- 16271012 TI - The role of genotyping in diagnosing cardiac channelopathies : progress to date. AB - The role of genotyping for diagnosis of the cardiac ion channelopathies is a work in progress. No formal guidelines or other publications discussing current recommendations for genotyping exist, particularly for clinical/commercial genotyping. Further, the field is changing rapidly, opinions vary and, additionally, circumstances inside the US are different from outside. The following considerations are a current summary based on a review of the literature, discussions with experts in the field, and our own opinions and also include a brief discussion about genotyping for therapeutic decision making. Research-based genotyping is very important for continued understanding of the details of pathophysiology and the complex regulatory processes in these diseases. Clinical/commercial genotyping for diagnosis is important for identifying patients with reduced penetrance of the phenotype since effective therapies to prevent sudden death exist. Clinical genotyping for therapeutic advantage has limited application at present but will become much more important if and when genotype-/mutation-type specific therapies are shown to be effective. The recommendations will progressively change as new research findings and new genotyping technologies appear. PMID- 16271013 TI - Applications of AmpliChip CYP450. AB - Pharmacogenetics has assumed increasing importance with the developing concepts of personalized medicine. There is a need to determine the metabolic status of an individual when using drugs, the actions of which are influenced by drug metabolizing enzymes. Cytochrome P450 (CYP) and its variants, particularly CYP2D6 and CYP2C19, play a role in the metabolism of approximately 25% of all prescription drugs. This review covers the role of the CYP system not only in the metabolism of drugs but also in the pathophysiology of disease. Various technologies for the assessment of CYP status are described, with the focus on AmpliChip CYP450 (Roche Molecular Diagnostics, Alameda, CA, USA), the first approved microarray molecular diagnostic test for the analysis of 29 polymorphisms and mutations of the CYP2D6 gene, and two polymorphisms of the CYP2C19 gene. It combines Roche's PCR technology with the GeneChip microarray system (Affymetrix, Santa Clara, CA, USA). Examples of numerous drugs that are metabolized by the CYP system are listed, and categories of antidepressants, antipsychotics, immunosuppressive and anticancer drugs are described to illustrate the role of testing for CYP polymorphisms in the therapeutic use of these drugs. CYP testing has applications in toxicology and absorption, distribution, metabolism and excretion (ADME) profiling as a guide to drug development. AmpliChip CYP450 may be used in conjunction with pharmacotherapy to guide decision making about selection of drugs and dosage. The test is not a solitary tool to determine optimum drug dosage, but is meant for use along with clinical evaluation and other methods for the selection of the treatment that is best suited for an individual patient. AmpliChip CYP450 is the first DNA microarray test to be cleared by the US FDA, and its clearance paves the way for similar microarray-based diagnostic tests to be developed in the future. This will facilitate the development of personalized medicine. PMID- 16271014 TI - Molecular basis for advances in cervical screening. AB - Human papillomaviruses (HPVs) cause cervical lesions, which can, in some instances, progress to high-grade neoplasia and cancer. Around half a million cases of cervical cancer occur each year, with most occurring in developing countries where cervical cancer is a major cause of cancer-related death. The reduction in cervical cancer incidence in developed countries is largely attributed to the introduction of cervical screening. Cervical screening currently depends on the identification by cytology of abnormalities in cells taken from the surface of the cervix. The standard Pap test was developed >50 years ago, and despite modifications, still forms the basis of the test currently in use in most routine screening laboratories. Advances in our understanding of the molecular mechanisms that lead to the development of cervical cancer have been slow to impact on screening, despite the relatively high false-negative rates that can be associated with the conventional Pap smear. Improvements in screening strategies fall into a number of categories. Methods that improve cell presentation and attempt to eliminate artefacts/obscuring debris can be combined with image analysis systems in order to enhance diagnostic accuracy. Such approaches still rely on cytological evaluation and do not incorporate advances in our knowledge of how HPV causes cancer. By contrast, markers of virus infection or cell cycle entry, particularly those that offer some degree of prognostic significance, may be able to highlight abnormal cells more reliably than cytology, and could be combined with cytology to improve the detection rate. Our understanding of the molecular biology of HPV infection and the organization of the HPV life-cycle during cancer progression provides a rational basis for marker selection. The general assumption that persistent active infection by high risk HPV types is the true precursor of cervical cancer provides the rationale for HPV DNA testing in conjunction with enhanced cytology, while the development of RNA-based approaches should allow active infections to be distinguished from those that are latent. The detection in superficial cells of marker combinations at the level of RNA or protein has the potential to predict disease status more precisely than the detection of markers in isolation. There is also a need for better prognostic markers if the predictive value of screening is to be improved. The potential to control infection by vaccination should reduce the incidence of HPV-associated neoplasia in the population, and this may cause a change in the way that screening is carried out. Nevertheless, the lack of a therapeutic vaccine, and the difficulties associated with eliminating infection by multiple high-risk HPV types, means that some form of screening will still be required as a preventive measure for the control of cervical cancer for the foreseeable future. PMID- 16271015 TI - Nonsense-associated alternative splicing of the human thyroglobulin gene. AB - INTRODUCTION: We have described in previous articles a nonsense mutation (4588C>T, R1511X) in exon 22 of the thyroglobulin (TG) gene in a member of a family with a complex history of congenital goiter. In the mutated thyroid gland, full-length thyroglobulin mRNA is almost undetectable. However, a smaller transcript is detected in which the mutated exon 22 is skipped and the reading frame restored. It is conceivable that alternative splicing might be a mechanism involved in the rescue of nonsense mutations. METHODS: To investigate whether the detection of the alternative mRNA is due to an increase in its concentration or its preferential amplification during reverse transcriptase-PCR in the absence of the normal full-length mRNA competitor, we set up an assay in which the competitor mRNA was provided. We also studied the effect of the 4588C>T mutation on exon definition and processing using wild-type and mutated minigenes. RESULTS: The detection of the alternative mRNA lacking exon 22 is not caused by the absence of the full-length competitor. In contrast, our results demonstrate that the alternative transcript preferentially accumulates in the mutated thyroid at a level similar to the full-length transcript in control tissue. Transient expression experiments with wild-type and mutated minigenes indicate that the mutated exon is as efficiently spliced as the wild-type, suggesting that the 4588C>T mutation does not interfere with exon 22 definition and processing. CONCLUSIONS: The alternative splicing of the TG gene described in this article constitutes a new case of nonsense-associated alternative splicing. We have shown that the mutation itself does not interfere with exon definition and processing in vitro. Our results support the hypothesis that the alternative splicing of the mutated exon is driven by the interruption of the reading frame. PMID- 16271016 TI - Multiplex-PCR assay for the deletions causing hereditary persistence of fetal hemoglobin. AB - INTRODUCTION: Hereditary persistence of fetal hemoglobin (HPFH) is a benign condition caused by the failure of normal switching from the fetal to the adult beta-globin gene, resulting in continuous production of fetal hemoglobin beyond the perinatal period. To date, eight deletions of variable size and position have been reported for HPFH. Southern hybridization and PCR are the most common methods used to detect each deletion. AIM: Our aim was to develop a multiplex-PCR assay to detect these deletions in a single tube in order to facilitate rapid and accurate molecular diagnosis. METHODS AND RESULTS: This report is the first application of multiplex-gap-PCR to detect all HPFH deletions simultaneously to expedite diagnosis. The deletion breakpoints were precisely identified for each deletion and primers were designed in the unique regions across the breakpoints of HPFH-1 (Black), HPFH-2 (Ghanaian), HPFH-3 (Asian Indian), HPFH-4 (Italian), HPFH-5 (Italian), HPFH-6 (Vietnamese), HPFH-7 (Kenyan), and SEA-HPFH (Southeast Asian). As many as 16 primers were used in a single amplification reaction by adjusting the relative primer concentrations. The multiplex-PCR approach was standardized on known positive control samples. We identified unique deletion specific products for each deletion. The results were confirmed by sequence analysis. CONCLUSIONS: We conclude that our multiplex-gap PCR strategy provides the most rapid and accurate diagnosis for the deletions in the beta-globin gene cluster causing HPFH. PMID- 16271011 TI - An overview of hip injuries in running. AB - Running has steadily gained in worldwide popularity and is the primary exercise modality for many individuals of all ages. Its low cost, versatility, convenience and related health benefits appeal to men and women of broad cultural, ethnic and economic backgrounds. With more children and adults participating in recreational and competitive running, the incidence of injuries has steadily increased. Most running-related injuries affecting the lower extremities are due to preventable training errors, and some may necessitate medical evaluation or a significant reduction in training. Hip injuries in runners are due to interactions of intrinsic and extrinsic factors that adversely affect the complex regional anatomy. Acute or chronic hip pain presents a diagnostic and therapeutic challenge because the vague, nonspecific symptoms and signs may originate from local, regional or distant foci. Muscle strains and tendonitis are the most common aetiologies of hip pain and typically result from sudden acceleration/deceleration manoeuvres, direction changes or eccentric contractions. Apophysitis and avulsion fractures may affect younger runners and produce localised pain at muscle attachment sites. Iliotibial band syndrome is a common cause of lateral hip and knee symptoms characterised by sharp or burning pain that is exacerbated by activity. Bursitis, due to repetitive activity or acute trauma, may affect the trochanteric, ischial or iliopectineal bursae. Hip osteoarthritis may also produce persistent pain that worsens with running. Stress fractures are potentially serious conditions that affect women more frequently than men. Snapping hip syndrome is a benign condition that results from tight connective tissues' passing repeatedly over the greater trochanter, anterior hip capsule, lesser trochanter, femoral head or iliopectineal eminence. Acetabular labral tears, sports hernias and nerve entrapment syndromes are also potential causes of persistent hip pain in runners. Treatment of hip pain in running should focus not only on addressing the symptoms but also identifying the underlying conditions that precipitated the injury. Injury prevention and comprehensive rehabilitation are essential, since prior hip injuries increase the risk of subsequent ones. Coaches, trainers and medical personal who care for runners should advocate running regimens, surfaces, shoes, technique and individualised conditioning programmes that minimise the risk of initial or recurrent hip injuries. PMID- 16271017 TI - Skewed X inactivation of the normal allele in fully mutated female carriers determines the levels of FMRP in blood and the fragile X phenotype. AB - BACKGROUND: The variable phenotype in female carriers of a full mutation is explained in part by non-random X-chromosome inactivation. The molecular diagnosis of fragile X syndrome is based on the resolution of the number of CGG triplet repeats and the methylation status of a critical CpG in the fragile X mental retardation gene (FMR1) promoter. Neighboring CpGs in the FMR1 promoter are supposed to be equally methylated or unmethylated. METHOD: Southern blot analysis was performed with double digestion, either with EcoRI/EagI or with HindIII/SacII. The EagI restriction site was studied by sequencing. The fragile X encoded protein (FMRP) was detected in white blood cells by Western blot. The fragile X phenotype was evaluated by specific clinical examinations. RESULTS: Within one family we found three female carriers of a full mutation and a different degree of methylation of the normal allele that correlated with the levels of FMRP in blood and the fragile X phenotype. Complete methylation at the EagI CpG target (but partially methylated SacII CpG site) was associated with extremely skewed X inactivation (confirmed by analysis of the methylation status at the PGK locus), undetectable FMRP in blood, and a male-like phenotype. CONCLUSIONS: In fully mutated female carriers the methylation status at the EagI restriction site correlates with the levels of FMRP in blood and the fragile X phenotype. Neighboring CpG sequences in the FMR1 promoter can be differentially methylated, which should be taken into consideration for molecular diagnosis. PMID- 16271022 TI - Arabinogalactan proteins: involvement in plant growth and morphogenesis. AB - Arabinogalactan proteins (AGPs) are highly glycosylated hydroxyproline-containing variously located proteoglycans dynamically regulated in the course of plant ontogenesis. Special functions of AGPs are still unclear, but their involvement in vegetative growth and reproduction of plants is well established. This review considers data on the structure, biosynthesis, and metabolism of AGPs. Special attention is given to involvement of AGPs in growth and morphogenesis, and possible mechanisms of their regulatory action are considered. AGPs are also compared with animal proteoglycans. PMID- 16271023 TI - Computer-aided analysis of spatial structure of some hydrolytic enzymes. AB - Using the MolScript version 2.1 computer program for protein molecule modeling and X-ray structure analysis data the spatial structures of several hydrolytic enzymes have been compared. These include glucoamylase from Aspergillus awamori and Saccharomycopsis fibuligera and lipases from Rhizopus japonicus. Results on homology of amino acid sequences and topology of secondary structure elements were obtained. 3D models of these enzymes with positioning of functionally important groups in the active site cavity were built. PMID- 16271024 TI - Effect of "external" superoxide anion on apoptosis in coleoptiles of wheat seedlings. AB - A derivative of phthalic acid, dibutylphthalate (DBP), which has gametocidal effect at the concentration of approximately 10(-4) M, increased apoptosis in coleoptiles of wheat seedlings. This was associated with activation of chromatin margination and generation of mitochondria-containing vesicles. At the same concentration, DBP activated the release by the coleoptiles of superoxide anion into the environment. Lower (10(-5) M) and higher (10(-3) M) concentrations of DBP virtually had no effect on either process. A probable mechanism of effect of the "external" superoxide anion on apoptosis within the plant cell is discussed. PMID- 16271025 TI - Molecular characteristics of OmpF-like porins from pathogenic Yersinia. AB - Nonspecific pore-forming proteins (porins) are the major proteins of the outer membrane of Gram-negative bacteria responsible for diffusion of low-molecular weight compounds. Nucleotide sequences of the OmpF-like porins from the pathogenic bacteria Yersinia pseudotuberculosis (YPS) and Yersinia enterocolitica (YE) were cloned and determined. Values of molecular weights (MW) and isoelectric points (IEP) calculated for these proteins (for OmpF-YPS: MW 37.7 kD, IEP 4.45; for OmpF-YE: MW 39.5 kD, IEP 4.34) are in good agreement with experimental data. The OmpF-like Yersinia porins are highly homologous to each other (83-92%) and also to the OmpF protein from Serratia marcescens (70%); the homology to the OmpF porin from E. coli is significantly lower (52-58%). Multiple alignment of the amino acid sequences of mature OmpF proteins provided the distribution of conservative amino acid residues typical for porins. Moreover, the OmpF-like porins from Yersinia are characterized by the presence of extended regions with high and low homologies, which coincide with the transmembrane domains and "external" loops, respectively, of the topological model of the OmpF porin from E. coli. By predictive methods, the secondary structure of the OmpF-like porins from Yersinia was obtained. This structure is represented by 16 beta-strands connected by short "periplasmic" and longer "external" loops with unordered structure. PMID- 16271026 TI - Functional role of the N-terminal domain of bacteriophage T4 gene product 11. AB - Bacteriophage T4 late gene product 11 (gp11), the three-dimensional structure of which has been solved by us to 2.0 A resolution, is a part of the virus' baseplate. The gp11 polypeptide chain consists of 219 amino acid residues and the functionally active protein is a three-domain homotrimer. In this work, we have studied the role of gp11 N-terminal domain in the formation of a functionally active trimer. Deletion variants of gp11 and monoclonal antibodies recognizing the native conformation of gp11 trimer have been selected. Long deletions up to a complete removal of the N-terminal domain, containing 64 residues, do not affect the gp11 trimerization, but considerably change the protein structure and lead to the loss of its ability to incorporate into the baseplate. However, the deletion of the first 17 N-terminal residues results in functionally active protein that can complete the 11(-)-defective phage particles in in vitro complementation assay. This region of the polypeptide chain is probably essential for gp11-gp10 stable complex formation at the early stages of phage baseplate assembly in vivo. A study of the gp10 deletion variants suggests that the central domain of gp10 trimer is responsible for the interaction with gp11. PMID- 16271027 TI - Formation of bacteriochlorophyll form B820 in light harvesting 2 complexes from purple sulfur bacteria treated with dioxane. AB - Treatment of some sulfur bacteria (Allochromatium minutissimum, Thiorhodospira sibirica, and Ectothiorhodospira halovacuolata WN22) with dioxane results in formation of the bacteriochlorophyll form B820 in the light harvesting complex LH2. This form characterized by absorption maximum at 820 nm has the same absorption spectrum as B820 subcomplex from LH1 complex. Appearance of the B820 form was accompanied by a sharp decrease in absorption in the carotenoid region. This phenomenon observed in all LH2 complexes investigated may be attributed to formation of colorless carotenoid aggregates. This is very similar to the previously reported dissociation of the LH1 complex with carotenoids into B820 subcomplexes. Although the B820 form corresponded the bacteriochlorophyll dimer, its circular dichroism spectrum showed that pigment molecules in this dimer exhibit different interaction than those in the B820 subcomplex. The dioxane treatment of LH2 complexes isolated from Rhodopseudomonas palustris bacteria grown under normal or low intensity illumination did not result in formation of such dimers. It is suggested that bacteriochlorophyll B820 formation is related to unique structure of LH2 complexes from the sulfur bacteria. PMID- 16271028 TI - Location of the bases modified by M.BcoKIA and M.BcoKIB methylases in the sequence 5 -CTCTTC-3 /5 -GAAGAG-3. AB - The strain Bacillus coagulans K contains two DNA-methyltransferases, M.BcoKIA and M.BcoKIB, which recognize the sequence 5 -CTCTTC-3 /5 -GAAGAG-3 and possess N4 methylcytosine and N6-methyladenine specificities, respectively. A special construct containing the recognition site of BcoKI and sites of four IIS restriction endonucleases (IIS restriction endonuclease cassette) was designed to locate the nucleotides modified by the methylases. The modified bases were determined as: 5 -m(4)CTCTTC-3 /5 -GAAGAm(6)G-3 . PMID- 16271029 TI - Effect of calcium ions on enteropeptidase catalysis. AB - The effects of calcium ions on hydrolysis of low molecular weight substrates catalyzed by different forms of enteropeptidase were studied. A method for determining activity of truncated enteropeptidase preparations lacking a secondary trypsinogen binding site and displaying low activity towards trypsinogen was developed using N-alpha-benzyloxycarbonyl-L-lysine thiobenzyl ester (Z-Lys-S-Bzl). The kinetic constants for hydrolysis of this substrate at pH 8.0 and 25 degrees C were determined for natural enteropeptidase (K(m) 59.6 microM, k(cat) 6660 min(-1), k(cat)/K(m) 111 microM(-1) x min(-1)), as well as for enteropeptidase preparation with deleted 118-783 fragment of the heavy chain (K(m) 176.9 microM, k(cat) 6694 min(-1), k(cat)/K(m) 37.84 microM(-1) x min(-1)) and trypsin (K(m) 56.0 microM, k(cat) 8280 min(-1), k(cat)/K(m) 147.86 microM(-1) x min(-1)). It was shown that the enzymes with trypsin-like primary active site display similar hydrolysis efficiency towards Z-Lys-S-Bzl. Calcium ions cause 3 fold activation of hydrolysis of the substrates of general type GD(4)K-X by the natural full-length enteropeptidase. In contrast, the hydrolysis of substrates with one or two Asp/Glu residues at P2-P3 positions is slightly inhibited by Ca2+. In the case of enteropeptidase light chain as well as the enzyme containing the truncated heavy chain (466-800 fragment), the activating effect of calcium ions was not detected for all the studied substrates. The results of hydrolysis experiments with synthetic enteropeptidase substrates GD(4)K-F(NO(2))G, G(5)DK F(NO(2))G (where F(NO(2)) is p-nitrophenyl-L-phenylalanine residue), and GD(4)K Nfa (where Nfa is beta-naphthylamide) demonstrate the possibility of regulation of undesired side hydrolysis using natural full-length enteropeptidase for processing chimeric proteins by means of calcium ions. PMID- 16271030 TI - Orientation and mobility of actin in different intermediate states of the ATP hydrolysis cycle. AB - Using polarization fluorimetry, we have investigated conformational changes of FITC-phalloidin-labeled F-actin in ghost muscle fibers. These changes were induced by myosin subfragment-1 (S1) in the absence and presence of MgADP, MgAMP PNP, MgATPgammaS, or MgATP. Modeling of various intermediate states was accompanied by discrete changes in actomyosin orientation and mobility of fluorescent dye dipoles. This suggests multistep changes of orientation and mobility of actin monomers during the ATPase cycle. The most pronounced differences in orientation (~4 degrees ) and in mobility (~43%) of actin were found between the actomyosin states induced by MgADP and MgATP. PMID- 16271031 TI - Effect of nucleotides on the orientation and mobility of myosin subfragment-1 in ghost muscle fiber. AB - Using polarization fluorimetry, the orientation and mobility of 1,5-IAEDANS specifically bound to Cys707 of myosin subfragment-1 (S1) were studied in ghost muscle tropomyosin-containing fibers in the absence and in the presence of MgADP, MgAMP-PNP, MgATPgammaS, or MgATP. Modeling of various intermediate states was accompanied by discrete changes in actomyosin orientation and mobility of fluorescent dye dipoles. This suggests multistep changes in the structural state of the myosin head during the ATPase cycle. Maximal differences in the probe orientation by 4 degrees and its mobility by 30% were found between actomyosin states in the presence of MgADP and MgATP. It is suggested that interaction of S1 with F-actin induces nucleotide-dependent rotation of the whole motor domain of the myosin head or only the dye-binding site and also change in the head mobility. PMID- 16271032 TI - Thermodynamic characteristics of plasminogen activation by indirect activators. AB - Several indirect plasminogen (Pg) activators are known including streptokinase and the monoclonal antibody IV-Ic, whose mechanism of activation is well studied. To characterize thermodynamically the activation of Pg by streptokinase (SK) and the monoclonal antibody (mAB) IV-Ic, the activation energies were calculated for various reaction stages. Activation energy of 7.4 kcal/mol was determined for the interaction of the chromogenic substrate S-2251 with plasmin (Pm) and activated equimolar complexes Pm-SK and Pg*SK at the steady-state reaction stage, and 18.7 kcal/mol with the complexes Pg*IV-Ic. A 2.5-fold increase in the energy of activation for the Pg*IV-Ic complex suggests a more intricate mechanism of its interaction with the substrate. At the stage of increasing active center concentrations and the formation of activated complexes Pg*SK and Pg*mAB IV-Ic, the activation energy was found to be 10.5 and 38 kcal/mol, respectively. At this reaction stage the conformational rearrangement of Pg molecule with the formation of active center is the limiting stage determining the reaction rate. Unexpectedly high energy of activation at the second stage of interaction between mAB IV-Ic and Pg suggests several simultaneous reactions and complexity of conformation rearrangement in the Pg molecule in activated complexes, thus requiring large energy expense. Formation of the active center is probably accompanied by its transition within a narrow temperature range into another conformation state with the change in activation parameters of the reaction. Quantitative evaluation of the studied reactions from the perspective of thermodynamics of the enzymatic reactions gives more comprehensive characteristics of the activation mechanism. PMID- 16271033 TI - Binding sites for transcription factor SF-1 in promoter regions of genes encoding mouse steroidogenesis enzymes 3betaHSDI and P450c17. AB - Using gel retardation of DNA samples and specific antibodies, binding sites for the transcription factor SF-1 were found in positions -53/-44 and -285/-270 in the promoter region of the mouse Cyp17 gene and in position -117/-108 of the promoter region of the mouse 3betaHSDI gene. PMID- 16271034 TI - Enhanced activity of DNA polymerase iota in mouse brain cells is associated with aggressiveness. AB - Recent studies performed with crude extracts of mouse tissues showed that the activity of DNA-polymerase iota (Pol iota) can be detected only in brain and testis extracts. To assess whether the activity of Pol iota is associated with animal behavior, we determined Pol iota activity in brain extracts of mice of two lines sharply differing in aggressiveness (RSB and RLB). We found that Pol iota activity in the mice with aggressive behavior was three times higher than in the less aggressive mice. The possible relationship between the activity of Pol iota and animal behavior is discussed. PMID- 16271036 TI - Role of two chloride-binding sites in functioning of testicular angiotensin converting enzyme. AB - Modeling the structure of the C-domain of bovine angiotensin-converting enzyme revealed two putative chloride-binding sites. The kinetic parameters, K(m) and k(cat), of hydrolysis of the substrate Cbz-Phe-His-Leu catalyzed by the testicular (C-domain) enzyme were determined over a wide range of chloride concentrations. Chloride anions were found to be enzyme activators at relatively low concentrations, but they inhibit enzymatic activity at high concentrations. A general scheme for the effect of chloride anions on activity of the C-domain of bovine angiotensin-converting enzyme accounting for binding the "activating" and "inhibiting" anions is suggested. PMID- 16271035 TI - Further characterization of a sarcoplasmic serine proteinase from the skeletal muscle of white croaker (Argyrosomus argentatus). AB - A trypsin-type serine proteinase (WSP) was purified previously from the sarcoplasmic fraction of skeletal muscle of white croaker (Argyrosomus argentatus) by Yanagihara et al. ((1991) Nippon Suisan Gakaishi, 57, 133-142). However, further research on WSP was not carried out. In the present study, we determined the N-terminal amino acid sequence of this enzyme (27 amino acid residues), which revealed relatively high identity in the conserved region to other trypsin-type serine proteinases. Degradation action of WSP on neuropeptides is also reported in this manuscript. The results show that WSP only cleaves at the carboxyl side of Arg or Lys residue of the peptides, especially between dibasic amino acid residues such as Arg-Arg and Arg-Lys. PMID- 16271037 TI - Reactions of peroxynitrite and nitrite with organic molecules and hemoglobin. AB - In this work, the reactions of nitrite (NO2-) and peroxynitrite (ONOO-) with organic molecules as well as with hemoglobin (Hb) were examined and the potential interference with the detection of hydrogen peroxide and Hb was investigated. ONOO- at low concentrations (35-140 microM) induced a concentration-dependent oxidation of o-phenylenediamine and guaiacol, and this process can be improved by the addition of Hb in a concentration-dependent manner. This enhancing effect of Hb was possibly due to the formation of such highly reactive species as ferrylHb during the reaction of ONOO- and Hb. NO2- also oxidized the aromatic amine o phenylenediamine, but its efficiency was much lower than that of ONOO-. A 300 fold excess of NO2- over hydrogen peroxide inhibited the oxidation of Pyrogallol Red mediated by hydrogen peroxide and Hb, which was due in part to the reaction of NO2- with Hb ferryl species compound I and compound II and the phenoxyl radical. These data suggest that ONOO- and NO2- can interfere with the detection of hydrogen peroxide. The overestimation or underestimation of the hydrogen peroxide detected is dependent upon the organic molecule utilized for detection and the relative rate of NO2-, superoxide, and ONOO- generation. PMID- 16271038 TI - Codon 12 region of mouse K-ras gene is the site for in vitro binding of transcription factors GATA-6 and NF-Y. AB - Codon 12 of the K-ras gene is a generally recognized example of a mutational hot spot. By the approach of gel retardation and specific antibodies, a double stranded oligonucleotide corresponding to the codon 12 region of the mouse K-ras gene (from 20 to 50 bp with respect to the exon 1 start) was found to be a site for cooperative binding of the transcription factors GATA-6 and NF-Y. GATA-6 and NF-Y were selectively activated with lung carcinogens 3-methylcholanthrene and nitrosoethylurea in mice of strains susceptible to lung tumorigenesis but not in animals of resistant strains. The interaction of GATA-6 and NF-Y with the codon 12 region of the K-ras gene is suggested to be involved in the mechanism of lung carcinogenesis. PMID- 16271040 TI - Homology-model-guided site-specific mutagenesis reveals the mechanisms of substrate binding and product-regulation of adenosine kinase from Leishmania donovani. AB - Despite designating catalytic roles of Asp299 and Arg131 during the transfer of gamma-phosphate from ATP to Ado (adenosine) [R. Datta, Das, Sen, Chakraborty, Adak, Mandal and A. K. Datta (2005) Biochem. J. 387, 591-600], the mechanisms that determine binding of substrate and cause product inhibition of adenosine kinase from Leishmania donovani remained unclear. In the present study, employing homology-model-guided site-specific protein mutagenesis, we show that Asp16 is indispensable, since its replacement with either valine or arginine resulted in a >200-fold increase in K(m) (Ado) with a 1000-fold decrease in k(cat)/K(m), implying its critical importance in Ado binding. Even glutamate replacement was not tolerated, indicating the essentiality of Asp16 in the maintenance of steric complementarity of the binding pocket. Use of 2'or 3'-deoxygenated Ado as substrates indicated that, although both the hydroxy groups play important roles in the formation of the enzyme-Ado complex, the binding energy (DeltaDeltaG(B)) contribution of the former was greater than the latter, suggesting possible formation of a bidentate hydrogen bond between Asp16 and the adenosyl ribose. Interestingly, AMP-inhibition and AMP-binding studies revealed that, unlike the R131A mutant, which showed abrogated AMP-binding and insensitivity towards AMP inhibition despite its unaltered K(m) (Ado), all the Asp16 mutants bound AMP efficiently and displayed AMP-sensitive catalytic activity, suggesting disparate mechanisms of binding of Ado and AMP. Molecular docking revealed that, although both Ado and AMP apparently occupied the same binding pocket, Ado binds in a manner that is subtly different from AMP binding, which relies heavily on hydrogen-bonding with Arg131 and thus creates an appropriate environment for competition with Ado. Hence, besides its role in catalysis, an additional novel function of the Arg131 residue as an effector of product-mediated enzyme regulation is proposed. PMID- 16271039 TI - Unique translational modification of an invertebrate neuropeptide: a phosphorylated member of the adipokinetic hormone peptide family. AB - Separation of an extract of corpora cardiaca from the protea beetle, Trichostetha fascicularis, by single-step RP (reverse-phase)-HPLC and monitoring of tryptophan fluorescence resulted in two distinctive peaks, the material of which mobilized proline and carbohydrates in a bioassay performed using the beetle. Material from one of these peaks was; however, inactive in the classical bioassays of locusts and cockroaches that are used for detecting peptides belonging to the AKH (adipokinetic hormone) family. After enzymatically deblocking the N-terminal pyroglutamic acid (pGlu) residue in the peptide material and sequencing by Edman degradation, a partial sequence was obtained: (pGlu)-Ile-Asn-Met-Thr-Xaa-Gly-Trp. The complete sequence was deduced from ESI-MS(n) (electrospray ionization multi stage-MS); position six was identified as a phosphothreonine residue and the C terminus is amidated. The peptide, code-named Trifa-CC, was chemically synthesized and used in confirmatory experiments to show that the primary structure had been correctly assigned. To our knowledge, this is the first report of a phosphorylated invertebrate neuropeptide. Synthetic Trifa-CC co-elutes with the natural peptide, found in the gland of the protea beetle, after RP-HPLC. Moreover, the natural peptide can be dephosphorylated by alkaline phosphatase and the product of that reaction has the same retention time as a synthetic nonphosphorylated octapeptide which has the same sequence as Trifa-CC. Finally, synthetic Trifa-CC has hypertrehalosaemic and hyperprolinaemic biological activity in the protea beetle, but even high concentrations of synthetic Trifa-CC are inactive in locusts and cockroaches. Hence, the correct peptide structure has been assigned. Trifa-CC of the protea beetle is an unusual member of the AKH family that is unique in its post-translational modification. Since it increases the concentration of carbohydrates and proline in the haemolymph when injected into the protea beetle, and since these substrates are also used during flight, we hypothesize that Trifa-CC controls the mobilization of these metabolites in the protea beetle. PMID- 16271041 TI - Cysteine-accessibility analysis of transmembrane domains 11-13 of human concentrative nucleoside transporter 3. AB - hCNT3 (human concentrative nucleoside transporter 3) is a nucleoside-sodium symporter that transports a broad range of naturally occurring purine and pyrimidine nucleosides as well as anticancer nucleoside drugs. To understand its uridine binding and translocation mechanisms, a cysteine-less version of hCNT3 was constructed and used for cysteine-accessibility and permeant-protection assays. Cysteine-less hCNT3, with 14 endogenous cysteine residues changed to serine, displayed wild-type properties in a yeast expression system, indicating that endogenous cysteine residues are not essential for hCNT3-mediated nucleoside transport. A series of cysteine-substitution mutants spanning predicted TMs (transmembrane domains) 11-13 was constructed and tested for accessibility to thiol-specific reagents. Mutants M496C, G498C, F563C, A594C, G598C and A606C had no detectable transport activity, indicating that a cysteine substitution at each of these positions was not tolerated. Two functional mutants in putative TM 11 (L480C and S487C) and four in putative TM 12 (N565C, T557C, G567C and I571C) were partially inhibited by MTS (methanethiosulphonate) reagent and high concentrations of uridine protected against inhibition, indicating that TMs 11 and 12 may form part of the nucleoside translocation pathway. The lack of accessibility of MTS reagents to TM 13 mutants suggests that TM 13 is not exposed to the nucleoside translocation pathway. Furthermore, G567C, N565C and I571C mutants were only sensitive to MTSEA (MTS-ethylammonium), a membranepermeant thiol reagent, indicating that these residues may be accessible from the cytoplasmic side of the membrane, providing evidence in support of the predicted orientation of TM 12 in the current putative topology model of hCNT3. PMID- 16271042 TI - Oxidative folding of hirudin in human serum. AB - Human serum contains factors that promote oxidative folding of disulphide proteins. We demonstrate this here using hirudin as a model. Hirudin is a leech derived thrombin-specific inhibitor containing 65 amino acids and three disulphide bonds. Oxidative folding of hirudin in human serum is shown to involve an initial phase of rapid disulphide formation (oxidation) to form the scrambled isomers as intermediates. This is followed by the stage of slow disulphide shuffling of scrambled isomers to attain the native hirudin. The kinetics of regenerating the native hirudin depend on the concentrations of both hirudin and human serum. Quantitative regeneration of native hirudin in undiluted human serum can be completed within 48 h, without any redox supplement. These results cannot be adequately explained by the existing oxidized thiol agents in human serum or the macromolecular crowding effect, and therefore indicate that human serum may contain yet to be identified potent oxidase(s) for assisting protein folding. PMID- 16271043 TI - Endothelin-1-induced oxidative stress in DOCA-salt hypertension involves NADPH oxidase-independent mechanisms. AB - We have demonstrated recently [Callera, Touyz, Teixeira, Muscara, Carvalho, Fortes, Schiffrin and Tostes (2003) Hypertension 42, 811-817] that increased vascular oxidative stress in DOCA (deoxycorticosterone acetate)-salt rats is associated with activation of the ET (endothelin) system via ETA receptors. The exact source of ET-1-mediated oxidative stress remains unclear. The aim of the present study was to investigate whether ET-1 increases generation of ROS (reactive oxygen species) in DOCA-salt hypertension through NADPH-oxidase dependent mechanisms. Xanthine oxidase, eNOS (endothelial nitric oxide synthase) and COX-2 (cyclo-oxygenase-2) were also examined as potential ET-1 sources of ROS as well as mitochondrial respiration. DOCA-salt and control UniNX (uninephrectomized) rats were treated with the ETA antagonist BMS182874 (40 mg.day(-1).kg(-1) of body weight) or vehicle. Plasma TBARS (thiobarbituric acid reacting substances) were increased in DOCA-salt compared with UniNX rats. Activity of NADPH and xanthine oxidases in aorta, mesenteric arteries and heart was increased in DOCA-salt rats. BMS182874 decreased plasma TBARS levels without influencing NADPH and xanthine oxidase activities in DOCA-salt rats. Increased p22(phox) protein expression and increased p47(phox) membrane translocation in arteries from DOCA-salt by rats were not affected by BMS182874 treatment. Increased eNOS and COX-2 expression, also observed in aortas from DOCA-salt rats, was unaltered by BMS182874. Increased mitochondrial generation of ROS in DOCA salt rats was normalized by BMS182874. ETA antagonism also increased the expression of mitochondrial MnSOD (manganese superoxide dismutase) in DOCA-salt rats. In conclusion, activation of NADPH oxidase does not seem to be the major source of oxidative stress induced by ET-1/ETA in DOCA-salt hypertension, which also appears to be independent of increased activation of xanthine oxidase or eNOS/COX-2 overexpression. Mitochondria may play a role in ET-1-driven oxidative stress, as evidenced by increased mitochondrial-derived ROS in this model of hypertension. PMID- 16271044 TI - Emerging evidence for a similar role of glutamate receptors in the nervous and immune systems. AB - The role of glutamate receptors in synaptic transmission and excitotoxicity in the nervous system is well established. Recent evidence has emerged that glutamatergic mechanisms also exist in a wide variety of non-neuronal cells. In the case of thymocytes and lymphocytes, several types of glutamate receptor are expressed which can induce functional changes. This review focuses on the cellular function of NMDA-activated ionotropic and groups I and III metabotropic glutamate receptors in lymphocytes. Levels of exogenous and endogenous circulatory agonists and antagonists for lymphocyte glutamate receptors, notably homocysteine metabolites, are markedly increased in certain disease states and may be involved in disorders of the immune system. In addition to glutamate and aspartate, these compounds are active at glutamate receptors and increase the excitotoxic effects of glutamate in both neurons and lymphocytes. Increased levels of compounds acting at glutamate receptors may be risk factors for organ damage, for example in both heart and kidney disease. We conclude that glutamate is involved in signaling in immunocompetent cells and that the expression of both ionotropic and metabotropic glutamate receptors may have regulatory functions in immunocompetent cells, as well as in the nervous system. In addition, glutamate may serve as a signaling agent between the immune and nervous systems. PMID- 16271045 TI - The scaffolding protein PSD-95 interacts with the glycine transporter GLYT1 and impairs its internalization. AB - Recent evidence indicates that the glycine transporter-1 (GLYT1) plays a role in regulation of NMDA receptor function through tight control of glycine concentration in its surrounding medium. Immunohistochemical studies have demonstrated that, as well as being found in glial cells, GLYT1 is also associated with the pre- and postsynaptic aspects of glutamatergic synapses. In this article, we describe the interaction between GLYT1 and PSD-95 in the rat brain, PSD-95 being a scaffolding protein that participates in the organization of glutamatergic synapses. Mutational analysis reveals that the C-terminal sequence of GLYT1 (-SRI) is necessary for the transporter to interact with the PDZ domains I and II of PSD-95. This C-terminal tripeptide motif also seems to be involved in the trafficking of GLYT1 to the membrane, although this process does not involve PDZ proteins. GLYT1 is able to recruit PSD-95 to the plasma membrane, but it does not affect its clustering. However, the interaction stabilizes this transporter at the plasma membrane, blocking its internalization and producing a significant increase in the V(max) of glycine uptake. We hypothesize that PSD-95 might act as a scaffold for GLYT1 and NMDA receptors, allowing GLYT1 to regulate the concentrations of glycine in the micro-environment of NMDA receptors. PMID- 16271046 TI - Intracellular events mediating insulin-like growth factor I-induced oligodendrocyte development: modulation by cyclic AMP. AB - Insulin-like growth factor I (IGF-I) is a potent inducer of oligodendrocyte development and myelination. Although IGF-I intracellular signaling has been well described in several cell types, intracellular mechanisms for IGF-I-induced oligodendrocyte development have not been defined. By using specific inhibitors of intracellular signaling pathways, we report here that the MAPK and phosphatidylinositol 3-kinase signaling pathways are required for the full effect of IGF-I on oligodendrocyte development in primary mixed rat cerebrocortical cell cultures. The MAPK activation, but not the phosphatidylinositol 3-kinase activation, leads to phosphorylation of the cAMP response element-binding protein, which is necessary for IGF-I to induce oligodendrocyte development. cAMP, although it does not show any effect on oligodendrocyte development, has an inhibitory effect on IGF-I-induced oligodendrocyte development that is mediated by the cAMP-dependent protein kinase. Furthermore, cAMP also has an inhibitory effect on IGF-I-dependent MAPK activation. This is a cAMP-dependent protein kinase-independent effect and probably contributes to the cAMP action on IGF-I induced oligodendrocyte development. PMID- 16271048 TI - Neuronal K+/Cl- co-transporter (KCC2) transgenes lacking neurone restrictive silencer element recapitulate CNS neurone-specific expression and developmental up-regulation of endogenous KCC2 gene. AB - The K+/Cl- co-transporter KCC2 maintains the low intracellular chloride concentration required for fast synaptic inhibition and is exclusively expressed in neurones of the CNS. Here, we show that the KCC2 gene (alias SLC12a5) has multiple transcription start sites and characterize the activity of 6.8 kb of mouse KCC2 gene regulatory sequence (spanning 1.4 kb upstream from exon 1 to exon 2) using luciferase reporters. Overexpression of neurone-restrictive silencer factor repressed the reporter activity in vitro, apparently via a neurone restrictive silencer element (NRSE(KCC2)) within intron 1 of the mouse KCC2 gene. In transgenic mice, however, KCC2 reporters with or without deletion of the NRSE(KCC2) were expressed exclusively in neurones and predominantly in the CNS with a similar pattern and developmental up-regulation as endogenous KCC2. Moreover, a third transgene with just a 1.4-kb KCC2 promoter region lacking the NRSE(KCC2)-bearing intron 1 was still expressed predominantly in neural tissues. Thus, developmental up-regulation of the KCC2 gene does not require NRSE(KCC2) and the 1.4-kb KCC2 promoter is largely sufficient for neurone-specific expression of KCC2. PMID- 16271047 TI - Tetanus toxin fragment C fusion facilitates protein delivery to CNS neurons from cerebrospinal fluid in mice. AB - To improve protein delivery to the CNS following intracerebroventricular administration, we compared the distribution of a human Cu/Zn superoxide dismutase:tetanus toxin fragment C fusion protein (SOD1:TTC) in mouse brain and spinal cord with that of tetanus toxin fragment C (TTC) or human SOD1 (hSOD1) alone, following continuous infusion into the lateral ventricle. Mice infused with TTC or SOD1:TTC showed intense anti-TTC or anti-hSOD1 labeling, respectively, throughout the CNS. In contrast, animals treated with hSOD1 revealed moderate staining in periventricular tissues. In spinal cord sections from animals infused with SOD1:TTC, the fusion protein was found in neuron nuclear antigen-positive (NeuN+) neurons and not glial fibrillary acidic protein positive (GFAP+) astrocytes. The percentage of NeuN+ ventral horn cells that were co-labeled with hSOD1 antibody was greater in mice treated with SOD1:TTC (cervical cord = 73 +/- 8.5%; lumbar cord = 62 +/- 7.7%) than in mice treated with hSOD1 alone (cervical cord = 15 +/- 3.9%; lumbar cord = 27 +/-4.7%). Enzyme linked immunosorbent assay for hSOD1 further demonstrated that SOD1:TTC-infused mice had higher levels of immunoreactive hSOD1 in CNS tissue extracts than hSOD1 infused mice. Following 24 h of drug washout, tissue extracts from SOD1:TTC treated mice still contained substantial amounts of hSOD1, while extracts from hSOD1-treated mice lacked detectable hSOD1. Immunoprecipitation of SOD1:TTC from these extracts using anti-TTC antibody revealed that the recovered fusion protein was structurally intact and enzymatically active. These results indicate that TTC may serve as a useful prototype for development as a non-viral vehicle for improving delivery of therapeutic proteins to the CNS. PMID- 16271050 TI - Cross-seeding of wild-type and hereditary variant-type amyloid beta-proteins in the presence of gangliosides. AB - We investigated the molecular mechanism underlying the ganglioside-induced initiation of the assembly of wild and hereditary variant-type amyloid beta proteins, including Arctic-, Dutch-, and Flemish-type amyloid beta-proteins. We monitored the assembly of amyloid beta-protein by thioflavin-T assay, western blotting and electron microscopy. We also examined how externally added amyloid beta-protein assembles in a cell culture. The assembly of wild-, Arctic-, Dutch-, and Flemish-type amyloid beta-proteins were accelerated in the presence of GM1, GM1, GM3 and GD3 gangliosides. Notably, all of these amyloid beta-proteins accelerated the assembly of different type of amyloid beta-protein, following prior binding to a specific ganglioside. A specific-ganglioside-bound form of variant-type amyloid beta-protein was recognized by the antibody (4396C) specific to the GM1-ganglioside-induced altered conformation of wild-type amyloid beta protein. Moreover, the assembly of these amyloid beta-proteins in the presence of a specific ganglioside was markedly suppressed by coincubation with 4396C. This study suggests that cross-seeding can occur between wild and hereditary variant type amyloid beta-proteins despite differences in their amino acid sequences. PMID- 16271049 TI - Altered expression of neprilysin family members in the pituitary gland of sleep disturbed rats, an animal model of severe fatigue. AB - Alterations of the expression of some peptidases in the pituitary gland of a fatigued rat model were identified. Rats were kept in a cage filled with water to a height of 1.5 cm to disturb deep sleep. After 24-h sleep disturbance, expression of neutral endopeptidase 24.11 (neprilysin) mRNA was increased in the intermediate lobe of the pituitary gland, whereas the mRNA expression of another family member, damage-induced neuronal endopeptidase, which is normally expressed in a subgroup of anterior pituitary cells, was significantly suppressed. These alterations were demonstrated by RT-PCR, northern blotting and in situ hybridization. Other family members, such as neprilysin 2 and endothelin converting enzyme-1, did not show any change in mRNA expression. An increase of neprilysin mRNA expression was not seen in any other tissues of the sleep disturbed rats. The enzymatic activity of neprilysin was also increased in the pituitary. The augmentation of neprilysin expression and activity was prolonged as long as the sleep disturbance continued (up to 5 days), and returned to the basal level when rats were allowed to sleep freely. These results suggest that peptide processing and degradation in the pituitary may be an influential factor in fatigued states such as sleep disturbance. PMID- 16271051 TI - Unraveling thrombin's true microglia-activating potential: markedly disparate profiles of pharmaceutical-grade and commercial-grade thrombin preparations. AB - Microglia are the resident immune cells of the CNS. Brain injury triggers microglial activation, leading to proliferation, changes in antigenic profile, NO production and cytokine release. It is widely believed that serum factors inundating the injured tissue can prompt this activation, leading to long-term phenotypic changes. We and others have recently reported that commercial-grade preparations of thrombin, a serine protease known for its central function in blood coagulation, activate microglial cells. Recent findings, however, have called into question the involvement of thrombin itself in the induction of microglial cytokine release and led us to systematically re-investigate the ability of the protease to induce a broad spectrum of microglial activation parameters. We used a pharmaceutical-grade recombinant human thrombin (rh-thr) and compared it with a commercial-grade plasma-derived bovine thrombin (pb-thr) preparation that has been used extensively in the literature, including in our own earlier report. We investigated the effect of these two thrombin preparations on proliferation, NO production, interleukin-6 and tumour necrosis factor-alpha release, intracellular calcium signaling and cell surface expression of CD95 (Fas) and CD40. Pb-thr induced robust responses in all variables tested. In contrast, rh-thr triggered calcium signals and induced small but significant changes in the expression of cell surface antigens, but had no effect on proliferation, NO production or cytokine release. Control studies assured equivalent thrombin potencies and excluded both species-specific effects and endotoxin (lipopolysaccharide) contamination as possible causes of the disparity. Our results indicate a substantially more restricted role for thrombin itself in microglial activation than previously appreciated, but point to several potentially important co-stimulatory effects. In addition, these results suggest that previous studies examining thrombin's activation of microglia should be cautiously re-interpreted. PMID- 16271052 TI - Adenosine A2A receptors and metabotropic glutamate 5 receptors are co-localized and functionally interact in the hippocampus: a possible key mechanism in the modulation of N-methyl-D-aspartate effects. AB - Hippocampal metabotropic glutamate 5 receptors (mGlu5Rs) regulate both physiological and pathological responses to glutamate. Because mGlu5R activation enhances NMDA-mediated effects, and given the role played by NMDA receptors in synaptic plasticity and excitotoxicity, modulating mGlu5R may influence both the physiological and the pathological effects elicited by NMDA receptor stimulation. We evaluated whether adenosine A2A receptors (A(2A)Rs) modulated mGlu5R-dependent effects in the hippocampus, as they do in the striatum. Co-application of the A(2A)R agonist CGS 21680 with the mGlu5R agonist (RS)-2-chloro-s hydroxyphenylglycine(CHPG) synergistically reduced field excitatory postsynaptic potentials in the CA1 area of rat hippocampal slices. Endogenous tone at A(2A)Rs seemed to be required to enable mGlu5R-mediated effects, as the ability of CHPG to potentiate NMDA effects was antagonized by the selective A(2A)R antagonist ZM 241385 in rat hippocampal slices and cultured hippocampal neurons, and abolished in the hippocampus of A(2A)R knockout mice. Evidence for the interaction between A(2A)Rs and mGlu5Rs was further strengthened by demonstrating their co localization in hippocampal synapses. This is the first evidence showing that hippocampal A(2A)Rs and mGlu5Rs are co-located and act synergistically, and that A(2A)Rs play a permissive role in mGlu5R receptor-mediated potentiation of NMDA effects in the hippocampus. PMID- 16271053 TI - The blood-brain barrier transmigrating single domain antibody: mechanisms of transport and antigenic epitopes in human brain endothelial cells. AB - Antibodies against receptors that undergo transcytosis across the blood-brain barrier (BBB) have been used as vectors to target drugs or therapeutic peptides into the brain. We have recently discovered a novel single domain antibody, FC5, which transmigrates across human cerebral endothelial cells in vitro and the BBB in vivo. The purpose of this study was to characterize mechanisms of FC5 endocytosis and transcytosis across the BBB and its putative receptor on human brain endothelial cells. The transport of FC5 across human brain endothelial cells was polarized, charge independent and temperature dependent, suggesting a receptor-mediated process. FC5 taken up by human brain endothelial cells co localized with clathrin but not with caveolin-1 by immunochemistry and was detected in clathrin-enriched subcellular fractions by western blot. The transendothelial migration of FC5 was reduced by inhibitors of clathrin-mediated endocytosis, K+ depletion and chlorpromazine, but was insensitive to caveolae inhibitors, filipin, nystatin or methyl-beta-cyclodextrin. Following internalization, FC5 was targeted to early endosomes, bypassed late endosomes/lysosomes and remained intact after transcytosis. The transcytosis process was inhibited by agents that affect actin cytoskeleton or intracellular signaling through PI3-kinase. Pretreatment of human brain endothelial cells with wheatgerm agglutinin, sialic acid, alpha(2,3)-neuraminidase or Maackia amurensis agglutinin that recognizes alpha(2,3)-, but not with Sambucus nigra agglutinin that recognizes alpha(2,6) sialylgalactosyl residues, significantly reduced FC5 transcytosis. FC5 failed to recognize brain endothelial cells-derived lipids, suggesting that it binds luminal alpha(2,3)-sialoglycoprotein receptor which triggers clathrin-mediated endocytosis. This putative receptor may be a new target for developing brain-targeting drug delivery vectors. PMID- 16271055 TI - Recognition and management of Staphylococcus aureus toxin-mediated disease. AB - The ubiquitous human pathogen Staphylococcus aureus is capable of producing a formidable range of extracellular toxins that can have significant deleterious effects on the host. Toxic shock syndrome (TSS) results from infection or colonization with a strain of S. aureus that produces staphylococcal enterotoxin(s). The key features of TSS are widespread erythroderma occurring in association with profound hypotension and multiple organ dysfunction. As morbidity and mortality from TSS are appreciable, early recognition of TSS combined with intensive supportive management is critical. Staphylococcal foodborne disease (SFD) is caused by contamination of food during preparation or serving by preformed S. aureus enterotoxin(s). Symptom-onset is abrupt and the disease may be severe enough to warrant hospitalization, but it is usually self limiting and does not require specific antistaphylococcal therapy. Staphylococcal scalded skin syndrome (SSSS) results from colonization or infection with a strain of S. aureus that produces epidermolytic toxin(s). SSSS ranges in severity from trivial focal skin blistering to extensive, life-threatening exfoliation. This review discusses the epidemiology, pathogenesis, diagnosis, and management of TSS, SFD and SSSS. PMID- 16271056 TI - Epidemiology, clinical features and management of infections due to community methicillin-resistant Staphylococcus aureus (cMRSA). AB - Methicillin-resistant Staphylococcus aureus (MRSA) was initially confined to hospitals, but in the late 1970s appeared in the community in the USA, primarily among intravenous drug users. In the 1990s, community MRSA (cMRSA) strains appeared in multiple areas of the world, and spread extensively. Initially, there were problems with the definition of 'community-acquired', which was exacerbated by the fact that if a time-based definition was used without stratification for risk factors, patients with healthcare-associated MRSA would be counted. Some cMRSA strains have entered the hospital environment to cause outbreaks of infection, which has added to the difficulty in separating the two types. cMRSA strains usually possess genes for Panton-Valentine leukocidin (PVL), which is associated with furunculosis and necrotizing pneumonia, and sometimes possess other virulence genes such as those for toxic shock syndrome or exfoliative toxins. Antimicrobial resistance to commonly used topical and oral agents is now appearing in certain cMRSA strains, which is complicating therapy. While cMRSA strains are usually susceptible to most non-beta-lactam antimicrobials, there is a lack of clinical trial data indicating which drugs have superior clinical efficacy. DNA fingerprinting methods have become more sophisticated over the last decade, and have determined that cMRSA strains have probably arisen from virulent methicillin-susceptible strains, most likely by horizontal transfer of methicillin-resistance genes from coagulase negative staphylococci to S. aureus on a limited number of occasions, and these clones have spread extensively throughout the world by person-to-person transmission. In Australia, the dominant cMRSA clones are the Western Australia, Oceania and Queensland strains. PMID- 16271057 TI - Recognition and management of infections caused by vancomycin-intermediate Staphylococcus aureus (VISA) and heterogenous VISA (hVISA). AB - Vancomycin resistance in Staphylococcus aureus has recently emerged as an important clinical problem with implications for laboratory detection and clinical management of patients infected with resistant strains. To date, low level vancomycin resistance in the form of vancomycin-intermediate S. aureus (VISA) and heterogenous vancomycin-intermediate S. aureus (hVISA) have been more common, with only four cases of true vancomycin resistant S. aureus (VRSA) reported. This article reviews current knowledge about the epidemiology, clinical manifestations and optimal management of hVISA and VISA infections. VISA and hVISA have now been reported from many countries, and these strains tend to occur in patients with significant comorbidities and previous antibiotic exposure. Despite the difficulties in laboratory detection, there are increasing data linking VISA and hVISA to failure of glycopeptide antimicrobial therapy. Aggressive surgical intervention and non-glycopeptide-based antimicrobial therapy appears to improve outcomes for patients infected with these low-level vancomycin resistant strains. Clinicians and diagnostic laboratories need to be aware of VISA and hVISA as a clinical problem, and consider aggressive surgical debridement and non-glycopeptide-based therapy where infections with such strains are suspected or proven. PMID- 16271058 TI - Diagnosis and management of Staphylococcus aureus bacteraemia. AB - Staphylococcus aureus bacteraemia (SAB) is common. Around 8000 cases occur per year in Australia, of which 60% are hospital- or healthcare-associated. Risk factors for SAB include injectable drug use, haemodialysis, indwelling vascular catheters and immunosuppression. Metastatic infection develops in up to one-third of patients with SAB, with joints and heart valves being the most commonly affected sites. Community-acquisition,persistent fever, positive blood cultures after 48 h of treatment and the presence of embolic lesions correlate with the presence of complicated SAB (i.e. high risk of endocarditis and/or other metastatic complications). All patients require careful clinical evaluation to exclude endocarditis and other metastatic foci. Echocardiography,preferably transoesophageal echocardiography, should be performed to exclude endocarditis. Most patients with SAB, and all with features of complicated SAB, require prolonged intravenous antibiotic therapy (at least 4 weeks), but a subgroup with good prognostic features may be suitable for shorter intravenous therapy (2 weeks). Penicillinase-resistant penicillins (e.g.flucloxacillin) are the agents of choice for SAB with methicillin-sensitive strains. Vancomycin or first generation cephalosporins are alternatives but have lower antimicrobial activity than flucloxacillin. However, vancomycin remains the therapy of choice for SAB due to methicillin-resistant strains. Combination therapy with gentamicin may be useful for the first few days of treatment in selected patients, but otherwise there are few data to support the use of combination regimens in SAB. Newer agents such as linezolid and quinupristin/dalfopristin may have a role in selected patients, especially in SAB due to S. aureus strains with reduced susceptibility to vancomycin. PMID- 16271059 TI - Staphylococcus aureus infective endocarditis: diagnosis and management guidelines. AB - S. aureus infective endocarditis (SAIE) is a serious infection associated with considerable morbidity and mortality. There is evidence that the incidence of SAIE is increasing. As its clinical features are non-specific, SAIE must be suspected in every case of S. aureus bacteraemia, whether it is associated with an obvious source or not. The optimal antimicrobial agent(s) and duration of treatment for SAIE are currently not known, but on the basis of present evidence, a minimum of 2 weeks of antimicrobial therapy is recommended for 'right-sided' SAIE, a minimum of 4 weeks for uncomplicated 'left-sided' SAIE, and a minimum of 6 weeks for complicated 'leftsided' or prosthetic valve SAIE. Although there is no evidence to suggest that combination therapy with a cell-wall active agent (e.g. flucloxacillin) and an aminoglycoside decreases mortality in SAIE, combination therapy should be considered during the initial 3-5 days of therapy as it can shorten the duration of bacteraemia. In complicated or prosthetic valve SAIE, early and close liaison with cardiology and cardiothoracic surgery services is essential. Rapid identification and susceptibility testing of the infecting organism are important in determining the choice of definitive antimicrobial therapy. PMID- 16271060 TI - Antibiotics currently used in the treatment of infections caused by Staphylococcus aureus. AB - Staphylococcal infections are a common and significant clinical problem in medical practice. Most strains of Staphylococcus aureus are now resistant to penicillin, and methicillin-resistant strains of S. aureus (MRSA) are common in hospitals and are emerging in the community. Penicillinase-resistant penicillins (flucloxacillin, dicloxacillin) remain the antibiotics of choice for the management of serious methicillin-susceptible S. aureus (MSSA) infections, but first generation cephalosporins (cefazolin, cephalothin and cephalexin), clindamycin, lincomycin and erythromycin have important therapeutic roles in less serious MSSA infections such as skin and soft tissue infections or in patients with penicillin hypersensitivity, although cephalosporins are contra-indicated in patients with immediate penicillin hypersensitivity (urticaria, angioedema, bronchospasm or anaphylaxis). All serious MRSA infections should be treated with parenteral vancomycin or, if the patient is vancomycin allergic, teicoplanin. Nosocomial strains of MRSA are typically multi-resistant (mrMRSA), and mrMRSA strains must always be treated with a combination of two oral antimicrobials, typically rifampicin and fusidic acid, because resistance develops rapidly if they are used as single agents. Most community-acquired strains of MRSA in Australia and New Zealand are non multiresistant (nmMRSA), and lincosamides (clindamycin, lincomycin) or cotrimoxazole are the antibiotics of choice for less serious nmMRSA infections such as skin and soft tissue infections. New antibiotics such as linezolid and quinupristin/dalfopristin have good antistaphylococcal activity but are very expensive and should be reserved for patients who fail on or are intolerant of conventional therapy or who have highly resistant strains such as hVISA (heterogenous vancomycin-intermediate S aureus). PMID- 16271061 TI - Diagnosis and management of catheter-related bloodstream infections due to Staphylococcus aureus. AB - Intravenous catheters are essential to modern medical care but frequently cause complications, the most important of which is infection, commonly due to Staphylococcus aureus. It is estimated at least 3000 episodes of catheter-related bloodstream infection occur annually in Australia, and 9% to 25% of patients with such infections die. Infection rates vary depending on the type of device, with the lowest rates associated with peripherally inserted central catheters and highest rates with haemodialysis catheters. In febrile patients, the presence of an intravenous catheter should always prompt consideration of whether the line is the source, even if there is no exit site inflammation. If catheter-related infection appears likely, the line should be removed if possible. Either peripheral and line tip cultures, or timed cultures of blood drawn peripherally and through the line, should be taken. Empirical antibiotics should be aimed at S. aureus and aerobic Gram-negative organisms, and blood cultures should be repeated at 72 h. If S. aureus is grown, cure requires removal of the catheter, at least 14 days of parenteral therapy, and consideration of echocardiography (preferably transoesophageal). If the patient remains febrile for >72 h, blood cultures at 72 h grow S. aureus, or there is a prosthetic heart valve, the risk of endocarditis is high and 6 weeks of parenteral therapy should be given. Prevention requires an organized system of surveillance, with a strict policy on insertion of central lines in controlled conditions and regimented catheter care. The role of impregnated catheters in prevention remains controversial. PMID- 16271063 TI - Diagnosis and management of staphylococcal infections of vascular grafts and stents. AB - Staphylococcus aureus and coagulase-negative staphylococci are the commonest bacterial causes of both vascular graft and stent infections. Infection may occur either from direct implantation or haematogenous spread, and occurs in less than 1% of aortic grafts, 2% to 5% of inguinal grafts, and rarely in stents. Death or amputation is common following these infections despite aggressive treatment. Infection with S. aureus is usually more acute than with coagulase-negative staphylococci but both cause systemic symptoms. Inguinal infections usually cause localized swelling often with a sinus tract, bleeding or distal embolism. Aortic infections commonly present with abdominal discomfort retroperitoneal infection or a mass from a false aneurysm. Stent infections usually cause pain, swelling, erythema and circulation disturbances of the ipsilateral limb. The most useful investigations are blood cultures and computerized tomography or magnetic resonance imaging. These imaging techniques have a high sensitivity and specificity in advanced graft infections but these are considerably lower in low grade infections. Persistence of perigraft fluid beyond 3 months after surgery is suspicious of infection. Aggressive antimicrobial therapy is an important part of management but surgery is usually required to cure both graft and stent infections. Where the organisms are susceptible, high-dose beta-lactam therapy (e.g. flucloxacillin, dicloxacillin or a first-generation cephalosporin) plus low dose gentamicin are recommended initially. Some authorities add rifampicin after 3-5 days treatment, but this is controversial. Antimicrobial therapy can be stopped 4-6 weeks after surgery if arterial stump cultures are negative but should be continued long-term, and perhaps indefinitely, if they are positive. PMID- 16271062 TI - Diagnosis and management of staphylococcal infections of pacemakers and cardiac defibrillators. AB - Staphylococcal species, usually Staphylococcus aureus or Staphylococcus epidermidis, account for 70% to 95% of pacemaker and cardiac defibrillator infections. Infection limited to the generator pocket may cause pain, redness and swelling that is often accompanied by drainage or fistula formation. In this instance, the generator should be removed and reimplanted at another site as cure is rare with antimicrobial therapy alone. Infection of the leads usually tracks along the wire to include the endocardial surface and may involve the tricuspid valve and pocket. Clinical manifestations vary from mild chronic non-specific symptoms to septic shock with marked localizing signs. Septic embolization to the lungs is common and may cause cough, chest pain and shortness of breath that may be misdiagnosed. Blood culture and trans-oesophageal echocardiography (TOE) are the most important investigations.TOE has a sensitivity of >90%. Lead infection without vegetations may occur and these infections should be treated as for endocarditis. Antimicrobial therapy is an important part of treatment but lead infections are unlikely to cured unless the device is removed. Vancomycin is suitable as initial antimicrobial therapy as this covers both S. aureus and coagulase-negative staphylococci. Flucloxacillin, dicloxacillin or a first generation cephalosporin are preferred if the organism is sensitive. The addition of low-dose gentamicin may improve bacterial killing. The duration of antimicrobial therapy and timing of replacement of the device have not been determined but 2 weeks treatment before removal and 2-4 weeks treatment after replacement is commonly administered. PMID- 16271064 TI - Management of bone and joint infections due to Staphylococcus aureus. AB - The clinical presentation, investigation, treatment and prevention of osteomyelitis, septic arthritis, and prosthetic joint infections due to Staphylococcus aureus are discussed in this review. It is difficult to make evidence-based recommendations on the treatment of these infections, as very little high quality clinical evidence exists. Experimental evidence, case series and published expert opinion are reviewed and used to suggest the preferred treatment options in each type of infection. A combination of prolonged intravenous antibiotic therapy and surgery is generally required for cure of S. aureus bone and joint infections. The current standard antibiotic therapy for these infections is an intravenous beta-lactam agent. Newer antibiotic approaches, such as use of fluoroquinolones and rifampicin are reviewed, as are newer surgical approaches, such as attempted salvage of infected prosthetic joints. PMID- 16271065 TI - Diagnosis and management of Staphylococcus aureus infections of the skin and soft tissue. AB - Infections involving the skin and soft tissue are common and range from superficial, localized and sometimes self-limiting infections to deep, rapidly spreading and potentially life-threatening infections. Skin infections caused by Staphylococcus aureus include primary pyodermas, while those involving the soft tissues include cellulitis and pyomyositis. Surgical site infections and infections in intravenous drug users are also commonly caused by S. aureus. The severity of the infection determines the choice of treatment. There are few studies that have critically appraised the use of antibiotics in skin and soft tissue infections, and most guidelines are based on expert opinion. The beta lactam group of antibiotics are the mainstay of treatment for methicillin susceptible S. aureus infections. For methicillin-resistant S. aureus (MRSA) infections, both with community-acquired and hospital-acquired strains--which are becoming an increasing problem--the antibiotic choice is determined by local susceptibility patterns. Macrolides, clindamycin and cotrimoxazole are options for community-acquired MRSA, while vancomycin is reserved for treatment of infections caused by multiresistant MRSA strains and for patients with suspected endocarditis or severe sepsis. Although a number of the newer antibiotics such as linezolid and quinopristin/dalfopristin have been shown to have good activity against MRSA, these agents should only be used with specialist advice. PMID- 16271067 TI - Suppression of cancer phenotypes through a multifunctional actin-binding protein, calponin, that attacks cancer cells and simultaneously protects the host from invasion. AB - Quantitative and/or qualitative alteration of actin cytoskeletal molecules, involved in the regulation of cellular dynamic functions, should be intimately related with cancer phenotypes. Based on several lines of experimental evidence from our group, and others, this report proposes a strategy to simultaneously attack cancer cells and protect the host from cancer invasion, with one molecule. Calponin h1, an actin-stabilizing protein that is also intimately related to signal transduction, is very often suppressed in vascular smooth muscle cells of malignant human tumors and in mesothelial cells by coexisting cancer cells. We generated mice deficient for calponin h1, exhibiting fragility in blood vessels and peritoneal membranes. Hematogenous cancer metastasis occurred more easily in the calponin h1-deficient mice than in wild-type mice, and the peritoneal dissemination was extremely enhanced. The fragility was rescued by the exogenous introduction of the calponin h1 gene into mesothelial cells of the peritoneum. Furthermore, calponin h1 gene transfer into several transformed cell lines resulted in a suppression of malignancy. The peritoneal dissemination of intraperitoneally-injected B16-F10 cells was suppressed by the calponin h1 gene, given to target both cancer cells and the mesothelial cells of the host. The multifunctional nature of the molecule, as a machinery player of cytoskeleton and mediator of signal transduction, probably resulted in a favorable recipient discriminating effect on cancerous and normal cells. Thus, we believe that if we use adequate multifunctional molecules for therapy, it is possible to simultaneously suppress cancer phenotypes and protect normal cells from the attack of cancer cells. PMID- 16271066 TI - p73, a sophisticated p53 family member in the cancer world. AB - p73 belongs to a family of p53-related nuclear transcription factors that includes p53, p73 and p63. The overall structure and sequence homology indicates that a p63/p73-like protogene is the ancestral gene, whereas p53 evolved later in higher organisms. In accordance with their structural similarity, p73 functions in a manner analogous to p53 by inducing tumor cell apoptosis and participating in the cell cycle checkpoint control through transactivating an overlapping set of p53/p73-target genes. In sharp contrast to p53, however, p73 is expressed as two NH(2)-terminally distinct isoforms including transcriptionally active (TA) and transcriptionally inactive (DeltaN) forms. DeltaNp73, which has oncogenic potential, acts in a dominant negative manner against TAp73 as well as p53. p73 is induced to be stabilized in response to a subset of DNA-damaging agents in a way that is distinct from that of p53, and exerts its pro-apoptotic activity. Several lines of evidence suggest that p73 can induce tumor cell apoptosis in a p53-dependent and p53-independent manner. Some tumors exhibit resistance to the p53-dependent apoptotic program, therefore p73, which can induce apoptotic cell death by p53-independent mechanisms, is particularly useful. In this review, we discuss the regulatory mechanisms of p73 activity, and also the functional significance of p73 in the regulation of cellular processes including tumorigenesis, apoptosis and neurogenesis. PMID- 16271069 TI - Risk of p53 gene mutation in esophageal squamous cell carcinoma and habit of betel quid chewing in Taiwanese. AB - A recent report suggested that BQ (BQ) chewing significantly correlated with the occurrence of esophageal squamous cell carcinoma (ESCC) in Taiwanese. BQ chewing was shown to be associated with p53 mutation in oral cancers. However, the relationship between BQ chewing and p53 mutation in ESCC is unclear. Seventy-five primary ESCC patients were enrolled for mutational analysis of the p53 gene using polymerase chain amplification and direct sequencing of amplified product. Thirty seven mutations of the p53 gene were detected in 45.5% (34/75) of tumor specimens. These mutations significantly clustered in exon 5 (21/37) of the p53 gene. The incidence of p53 mutations did not associate with clinicopathological characteristics or the habits of cigarette smoking or alcohol consumption. However, BQ chewers exhibited significantly higher incidence of p53 gene mutations than non-chewers (67.6% vs 32.4%, P = 0.007). After controlling the confounding factors of cigarette smoking and alcohol intake, BQ chewing still showed significant association with the incidence of p53 mutation in ESCCs (RR = 4.23; 95% CI, 1.317-13.60). The A:T to G:C transition (8/37, 21.6%) and G:C to T:A transversion (5/23, 13.5%) were the prevalent spectrum of p53 gene mutations. All A:T to G:C transitional mutations occurred in patients with the habits of BQ chewing and cigarette smoking. Noticeably, alcohol consumption could enhance this peculiar spectrum of p53 mutation in ESCC. Accordingly, p53 might be an important molecular target of BQ carcinogens in the development of ESCC in Taiwanese. PMID- 16271068 TI - Dose-dependence of promotion of 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline induced rat hepatocarcinogenesis by ethanol: evidence for a threshold. AB - Although ethanol is thought to be a tumor-promoter, there are conflicting results concerning its effects on experimental hepatocarcinogenesis. Furthermore, the relationship between the amount of ethanol consumed and tumor promoting effects has hitherto not been investigated in detail. In the present study, 21-day-old F344/DuCrj rats were fed 200 p.p.m. 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) in their diet for 8 weeks and thereafter received ethanol at doses of 0, 0.1, 0.3, 1, 3, 10 and 20% in drinking water ad libitum for 16 weeks. The incidences of hepatocellular adenoma and total tumors increased dose-dependently with statistical significance at doses of 10% and 20%, compared to the initiated control value. Similarly, dose dependence was observed for the incidence of hepatocellular carcinoma, which was elevated significantly at the dose of 20%. No alteration in development of preneoplastic glutathione-S-transferase placental form positive foci or tumors was observed with 0.1-1%. Cell proliferation also increased dose-dependently and CYP2E1 protein induction was recognized in centrilobular regions without alteration in mRNA levels, but no effects were evident on formation of 8-hydroxy-2'-deoxyguanosine, an oxidative DNA damage marker, or lipid peroxidation in any of the initiated groups. The mRNA expression of cyclin D1 increased dose dependently. The results demonstrated that ethanol dose-dependently promotes hepatocarcinogenesis induced by MeIQx, but with no adverse influence at doses of 1% or less, comparable to sensible drinking levels in humans. PMID- 16271070 TI - Invasive human pancreatic carcinoma cells adhere to endothelial tri-cellular corners and increase endothelial permeability. AB - Although adhesive interactions between metastasizing cancer cells and vascular endothelial cells are critical in hematogenous metastasis, the early molecular events of the cancer-endothelial interaction remain largely obscure. Here we investigated the functional impact of cancer cells on endothelial permeability. We examined the binding of human pancreatic carcinoma cells MIA PaCa-2, PANC-1 and PSN-1 to a human umbilical vein endothelial cell (HUVEC) monolayer and the subsequent changes in the transendothelial electronic resistance (TEER) of the HUVEC. We found that MIA PaCa-2 and PANC-1 cells preferentially bound to the tri cellular corners of HUVEC and induced a rapid and irreversible reduction of TEER. The reduction of HUVEC TEER was associated with the focal disengagement of endothelial junctional adhesion molecules VE-cadherin and CD31. Blocking antibodies to integrin beta1, CD44, or CD9 affected neither the MIA PaCa-2 binding to HUVEC nor the reduction of TEER. Specific inhibitors for metalloproteinases, tyrosine-kinases and lipoxigenases, and a neutralizing anti vascular endothelial growth factor antibody failed to affect the MIA PaCa-2 induced reduction of HUVEC TEER, whereas treatment of the cells with paraformaldehyde or cytochalasin B abrogated the TEER reduction. These findings indicate that the MIA PaCa-2 cells bind selectively to endothelial tri-cellular corners, triggering a reduction of HUVEC TEER, which requires the active metabolism and intact actin cytoskeleton of the carcinoma cells, and is apparently unrelated to previously described cell adhesion and soluble factor pathways. Our data indicate a novel cell-contact-dependent mechanism for the cancer cell-mediated breakdown of endothelial barrier functions, which may be important in hematogenous cancer metastasis. PMID- 16271071 TI - Characterization of human NSCLC cell line with innate etoposide-resistance mediated by cytoplasmic localization of topoisomerase II alpha. AB - Topoisomerase (topo) II alpha is a target for many chemotherapeutic agents in clinical use. In tumor cells resistant to topo II poisons, there have been descriptions of quantitative and qualitative alterations involved in this enzyme. More recently, the cytoplasmic localization of topo II alpha has been described as a mechanism to confer drug resistance. Here, we report the characterization of a human non-small-cell lung cancer cell line, INER-37, which shows an innate resistance to etoposide. In this cell line, etoposide resistance was directly associated with the expression of topo II alpha resident mainly in the cytoplasmic region. At the molecular level, INER-37 cells carry on a heterozygous gene deletion, transcribing two different topo II alpha mRNAs: 4.8 kb and 2.0 kb. The bigger 4.8 kb mRNA (missing 1.3 kb of 3' mRNA and including the untranslated region) is translated into a truncated cytoplasmic protein of approximately 160 kDa. The protein truncation affects at least 96 amino acids in the COOH-terminal region where the more proximal bipartite nuclear localization signal is located. The INER-37 cell line is the first cancer cell line reported with an innate mutation affecting the 3'-end region of the topo II alpha gene that confers a cytoplasmic localization of the enzyme and therefore an increased resistance to etoposide. PMID- 16271072 TI - Targeting Id1 and Id3 inhibits peritoneal metastasis of gastric cancer. AB - Inhibitor of DNA binding (Id) proteins are essential for cell differentiation, proliferation, migration, invasion and angiogenesis. Recently, they have been shown to correlate with less differentiated phenotypes, high malignant potential and poor clinical outcome in various kinds of tumors. In an attempt to develop new strategies for the treatment of peritoneal metastasis of gastric cancer, we prepared an Id1, 3 double-knockdown gastric cancer cell line, MKN45, by RNA interference and investigated its effects on the development of metastatic nodules in the peritoneal cavity. Both cell proliferation and migration capabilities were decreased in Id1, 3 double-knockdown cells, as was their ability to bind to laminin, which could be explained by the decreased expression of integrin alpha6. These are important steps in the metastatic process. In a mouse model, the number of peritoneal metastatic nodules formed by Id1, 3 double knockdown cells was reduced compared to mock-transfected control cells, as was the size of individual tumors. In this study, we clearly demonstrated that Id1, 3 double-knockdown significantly impaired the ability of gastric cancer cells to form peritoneal metastasis. Id should be considered an ideal target for the treatment and prevention of gastric cancer, and RNA interference is an attractive and promising strategy to achieve it. PMID- 16271073 TI - The ALK-5 inhibitor A-83-01 inhibits Smad signaling and epithelial-to-mesenchymal transition by transforming growth factor-beta. AB - Transforming growth factor (TGF)-beta signaling facilitates tumor growth and metastasis in advanced cancer. Use of inhibitors of TGF-beta signaling may thus be a novel strategy for the treatment of patients with such cancer. In this study, we synthesized and characterized a small molecule inhibitor, A-83-01, which is structurally similar to previously reported ALK-5 inhibitors developed by Sawyer et al. (2003) and blocks signaling of type I serine/threonine kinase receptors for cytokines of the TGF-beta superfamily (known as activin receptor like kinases; ALKs). Using a TGF-beta-responsive reporter construct in mammalian cells, we found that A-83-01 inhibited the transcriptional activity induced by TGF-beta type I receptor ALK-5 and that by activin type IB receptor ALK-4 and nodal type I receptor ALK-7, the kinase domains of which are structurally highly related to those of ALK-5. A-83-01 was found to be more potent in the inhibition of ALK5 than a previously described ALK-5 inhibitor, SB-431542, and also to prevent phosphorylation of Smad2/3 and the growth inhibition induced by TGF-beta. In contrast, A-83-01 had little or no effect on bone morphogenetic protein type I receptors, p38 mitogen-activated protein kinase, or extracellular regulated kinase. Consistent with these findings, A-83-01 inhibited the epithelial-to mesenchymal transition induced by TGF-beta, suggesting that A-83-01 and related molecules may be useful for preventing the progression of advanced cancers. PMID- 16271074 TI - Identification of glioma-specific RFX4-E and -F isoforms and humoral immune response in patients. AB - For regulatory factor X4 (RFX4), two alternatively spliced variants, RFX4-A and B, were reported in the testis. In this study, we identified transcript variants RFX4-C, -D, -E, and -F, and demonstrated by reverse transcription-polymerase chain reaction (RT-PCR) that RFX4-A, -B and -C mRNAs were expressed only in the testis, and RFX4-D mRNA was expressed only in normal brain tissues. In tumors, RFX4-E and -F in addition to RFX4-D mRNA were expressed in gliomas by rapid amplification of cDNA ends and RT-PCR analyses. Expression of RFX4 mRNA was not observed in other tumors, such as lung, esophageal, stomach, colon or liver cancers. Quantitative real-time RT-PCR using common primer pairs detecting all of the variant transcripts showed high expression in normal testis, low expression in the brain (1% compared to the expression in testis), and overexpression in 17 of 61 gliomas (28%). Western blot analysis using DC28 monoclonal antibody (mAb) produced against recombinant RFX4-D C-terminus protein showed expression of RFX4 A and -C proteins, but not RFX4-B protein, in the testis, and expression of RFX4 D protein in the brain. Moreover, expression of RFX4-E and -F proteins, but not RFX4-D protein, was observed in gliomas. Immunohistochemistry analysis using DC28 mAb showed positive staining in the nuclei of spermatocytes in the testis and glioma cells. Antibody against RFX4 was detected in the sera of 3 of 58 (5%) glioma patients by enzyme-linked immunosorbent assay, suggesting the immunogenicity of RFX4-E and -F proteins in glioma patients. PMID- 16271075 TI - Serum level of soluble CD30 correlates with the aggressiveness of adult T-cell leukemia/lymphoma. AB - Adult T-cell leukemia/lymphoma (ATL) is a highly aggressive disease with poor prognosis. CD30(+) cells are frequently observed in lymph node cells and peripheral blood mononuclear cells of ATL patients. In order to elicit the role of CD30(+) cells in ATL development, we investigated expression of the membrane type of CD30 (mCD30) and the soluble form of CD30 (sCD30) on ATL cells. Both mCD30 and sCD30 are expressed on various numbers of ATL cells in vivo as well as cell lines such as MT-2, L540 and Karpas 299. The level of serum sCD30 in each clinical stage showed an elevated level in patients with acute type (mean +/- standard error; 545.2 +/- 18.6 U/mL) rather than with lymphoma type ATL (327.62 +/- 94.85 U/mL). In four patients whose sera were stored and examined longitudinally, the levels decreased following the response to chemotherapy but not in patients with chemotherapy resistance. Thus, our results imply that sCD30 levels may be another useful marker for the activity and aggressiveness of ATL. PMID- 16271077 TI - Involvement of mitochondrial aggregation in arsenic trioxide (As2O3)-induced apoptosis in human glioblastoma cells. AB - Arsenic trioxide (As(2)O(3)) is effective against acute promyelocytic leukemia and has potential as a novel treatment against malignant solid tumors. As(2)O(3) induces differentiation and inhibits growth. It also causes mitochondrial damage mediated by the production of reactive oxygen species (ROS) and the dissipation of mitochondrial transmembrane potential (DeltaPsi(m)), leading to apoptosis. Mitochondria might be the key target of antitumor activity by As(2)O(3); however, its mechanisms have not been completely elucidated. Using two human glioblastoma cell lines, A172 and T98G, we found that As(2)O(3) induced apoptosis in A172 cells but not in T98G cells. As(2)O(3)-induced ROS production was observed in both cell lines; however, the dissipation of DeltaPsi(m), Bax oligomerization and caspase activation occurred only in As(2)O(3)-sensitive A172 cells. To determine the mechanisms of As(2)O(3)-induced apoptosis after ROS generation, we examined the change of mitochondrial morphology. As we reported previously, mitochondrial aggregation occurs before cytochrome c release during apoptosis, thus playing a role in cell death progression. We observed mitochondrial aggregation in As(2)O(3)-sensitive A172 cells but not in T98G cells treated with As(2)O(3). Using laser scanning cytometry, we quantitatively confirmed the results, which indicate that mitochondrial aggregation plays an important role in regulating sensitivity to As(2)O(3)-induced apoptosis. We propose a sequential process involving ROS generation, mitochondrial aggregation, Bax oligomerization and DeltaPsi(m) dissipation, and caspase activation during As(2)O(3)-induced apoptosis. PMID- 16271076 TI - Leptosins isolated from marine fungus Leptoshaeria species inhibit DNA topoisomerases I and/or II and induce apoptosis by inactivation of Akt/protein kinase B. AB - DNA topoisomerases (topo) I and II are molecular targets of several potent anticancer agents. Thus, inhibitors of these enzymes are potential candidates or model compounds for anticancer drugs. Leptosins (Leps) F and C, indole derivatives, were isolated from a marine fungus, Leptoshaeria sp. as cytotoxic substances. In vitro cytotoxic effects of Lep were measured using 3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide-based viability assay. Lep F inhibited the activity of topos I and II, whereas Lep C inhibited topo I in vitro. Interestingly both of the compounds were found to be catalytic inhibitors of topo I, as evidenced by the lack of stabilization of reaction intermediate cleavable complex (CC), as camptothecin (CPT) does stabilize. Furthermore, Lep C inhibited the CC stabilization induced by CPT in vitro. In vivo band depletion analysis demonstrated that Lep C likewise appeared not to stabilize CC, and inhibited CC formation by CPT, indicating that Lep C is also a catalytic inhibitor of topo I in vivo. Cell cycle analysis of Lep C-treated cells showed that Lep C appeared to inhibit the progress of cells from G(1) to S phase. Lep C induced apoptosis in RPMI8402 cells, as revealed by the accumulation of cells in sub-G(1) phase, activation of caspase-3 and the nucleosomal degradation of chromosomal DNA. Furthermore, Leps F and C inhibited the Akt pathway, as demonstrated by dose-dependent and time-dependent dephosphorylation of Akt (Ser473). Our study shows that Leps are a group of anticancer chemotherapeutic agents with single or dual catalytic inhibitory activities against topos I and II. PMID- 16271078 TI - Abstracts from the 30th Annual Scientific Meeting of the Australasian Division of the International Academy of Pathology. June 3-5, 2005. Sydney, Australia. PMID- 16271079 TI - Side-to-side linking of myocardial cells in hypertrophic cardiomyopathy: whole heart microscopic observation with tangential sections. AB - By cross-section or longitudinal section, it is difficult to investigate longitudinal features of myocardial cells in the whole heart. Here, introducing the use of tangential sections to obtain longitudinal aspect of myocardial cells in any part of myocardium, the authors evaluated myocardium in the left ventricle in 10 normal hearts and four hearts with hypertrophic cardiomyopathy (HCM). Tangential sections were obtained by peeling the superficial layer of myocardium. After peeling the whole surface, secondary deep layer was peeled. These procedures were repeated more than five times through the wall. Intercalated discs (ICD) were observed immunohistochemically with anti-N-cadherin and antidesmoplakin. In normal hearts, myocardial cells were cut longitudinally and ran parallel in tangential sections. They linked end-to-end with simple and regular ICD with average lengths of 120-130 microm and average sarcomere numbers of 56-65. In HCM hearts, many myocardial cells were cut almost longitudinally running approximately parallel in tangential sections. Myocardial cells frequently showed side-to-side linking characterized by skewed ICD, indistinct ICD counterparts, and longitudinally arranged ICD. Two young HCM hearts had circle-shaped ICD and vacuole-like structures highlighted by immunostaining for N cadherin, which were actually extracellular structures comparable with irregular side-to-side linking. It is considered that side-to-side linking of myocardial cells is a characteristic microscopic feature in HCM rather than myocardial disarray. PMID- 16271080 TI - Expression of double-stranded RNA-activated protein kinase in small-size peripheral adenocarcinoma of the lung. AB - The authors investigated the protein expression of double-stranded RNA-activated protein kinase (PKR), which was identified by using a previous cDNA microarray study, to discover PKR's correlations with several pathological parameters and to elucidate its role in neoplastic transformation and progression of lung adenocarcinomas. Immunohistochemistry for PKR was performed and a semiquantitative scoring method was calculated based on staining intensity and percentage of immunoreactive tumor cells (high vs low) for one bronchioloalveolar carcinoma (BAC), 16 adenocarcinomas consisting of BAC and invasive carcinoma (mixed) and 21 invasive adenocarcinomas without BAC (invasive). The BAC had high grade expression and the mixed type tended to more frequently show high-grade expression than the invasive type (P = 0.028). There were no significant associations with age, tumor size, lymph node metastasis, lymphovascular invasion or the pathological stage. The Kaplan-Meier survival curves demonstrated that the patients with high-grade PKR expression had significantly shorter survival periods than those patients with low-grade PKR expression (P = 0.018). These results do not support the concept of PKR as a tumor suppressor in small-size peripheral adenocarcinomas of the lung. PMID- 16271082 TI - Computerized nuclear morphometry in thyroid follicular neoplasms. AB - Differential diagnosis of follicular adenoma (FA) and follicular carcinoma (FC) of the thyroid can be challenging in the routine practice of surgical pathology because the diagnosis of FC is strictly defined and identification depends on the presence of invasion of the capsule or blood vessels. These features may be equivocally presented in the histological sections and interpreted subjectively by different pathologists, so an objective approach to solve this problem is essential. Computerized morphometry is a scientific tool to evaluate cellular changes and it can enhance the interpretation of morphological features by the transformation of pathological changes in cells to a qualitative form. The present study investigated the diagnostic role of objective computerized nuclear morphometry in follicular neoplasms. Thirty-six cases of thyroid FC and 36 cases of FA from patients who were matched by age and sex were studied. Four nuclear parameters were selected and analyzed: mean nuclear area, mean nuclear perimeter, largest to smallest diameter ratio of the nuclei, and coefficient of variation of the nuclear area. The results indicate that all the chosen nuclear variables were significantly correlated with the FA and FC studied. In conclusion, computerized nuclear morphometry can be considered a helpful ancillary tool for differential diagnosis of FA and FC. PMID- 16271081 TI - Transcription factor-mediated proliferation and apoptosis in benign and malignant thyroid lesions. AB - Transcription factors play an essential role in regulating both cell proliferation and programmed cell death. Proliferation and apoptosis-related transcription factor immunoexpression patterns were concomitantly investigated in tissue sections of normal thyroid, goiters, follicular adenomas and well differentiated papillary and follicular carcinomas using antibodies against prothymosin alpha, E2F-1, p53, Bcl2, and Bax proteins. Proliferation and apoptotic indices were determined by Ki-67 immunoreactivity and the terminal deoxynucleotidyl transferase-mediated deoxy uridine triphosphate nick-end labeling technique, respectively. Prothymosin alpha and E2F-1 immunoexpression levels were found to be significantly elevated in well-differentiated carcinomas compared to adenomas, goiters and normal tissues (P < 0.05). Both proteins were directly correlated with the proliferation index (P < 0.05). E2F-1 was additionally correlated with the apoptotic index (P < 0.05). The majority of cases were negative for p53 staining. Positive Bcl2 immunostaining was detected in all thyroid histotypes. None of the normal tissues showed Bax immunoreactivity, while positive accumulation differed significantly between hyperplastic and neoplastic histotypes. Direct correlations were observed between prothymosin alpha and Bcl2 as well as between E2F-1 and Bax immunoexpression (P < 0.05). These data demonstrate that prothymosin alpha and E2F-1 are strongly involved in the proliferation processes of thyroid neoplasias. Furthermore, prothymosin alpha may promote cell survival through the Bcl2 anti-apoptotic pathway, while E2F-1-induced apoptosis via p53-independent pathways may be associated with transcriptional activation of bax pro-apoptotic gene. PMID- 16271083 TI - Increasing 14-3-3 sigma expression with declining estrogen receptor alpha and estrogen-responsive finger protein expression defines malignant progression of endometrial carcinoma. AB - 14-3-3 sigma (sigma) is a negative regulator of the cell cycle and contributes to G2 arrest. Lack of its expression due to hypermethylation of CpG islands has been reported in some carcinomas. A recent study showed that 14-3-3 sigma was down regulated through proteolysis by estrogen-responsive finger protein (Efp). Here, we investigated the expression of 14-3-3 sigma, hormone receptors, Efp and p53 in 86 cases of endometrial adenocarcinoma and 46 cases of normal or non-neoplastic endometria by means of immunohistochemistry and methylation-specific polymerase chain reaction. In normal endometrium, 14-3-3 sigma was overexpressed in the mid- to late-secretory phase due to hypomethylation. In endometrial adenocarcinoma, 14 3-3 sigma expression was low in low grade endometrioid adenocarcinoma due to hypermethylation, and increased significantly with increasing histological grade due to hypomethylation. 14-3-3 sigma expression inversely correlated with estrogen receptor alpha, progesterone receptor and Efp, and positively correlated with myometrial invasion and lymph node metastasis. These results suggest that 14 3-3 sigma was one of the menstrual cycle-related proteins regulated by epigenetic methylation, and its expression was influenced by epigenetic methylation or hormone receptors in progression of endometrial adenocarcinoma, and therefore was more than just a cell-cycle regulator. PMID- 16271085 TI - Cellular and stromal characteristics in the scirrhous hepatocellular carcinoma: comparison with hepatocellular carcinomas and intrahepatic cholangiocarcinomas. AB - Scirrhous hepatocellular carcinoma (SHCC) is a rare variation of HCC, for which characteristics of tumor cells and the fibrotic stroma have not been clarified in detail. The present study was therefore carried out to elucidate cytological features of tumor and stromal cells and components of the stromal extracellular matrix in 15 SHCC patients undergoing hepatectomy without preoperative transarterial embolization. Diagnosis was on the basis of a scirrhous histological pattern exceeding 50% of the tumor area. Expression of cytoplasmic and extracellular matrix proteins was compared among SHCC, HCC and intrahepatic cholangiocarcinoma (ICC) cases with immunohistochemical staining. The lesions could be histologically divided into radiating and sinusoidal types. Common stromal components of SHCC and ICC were collagen types I and III. There was no expression of laminin-5 in the stroma of SHCC, but it was present in almost all ICC cases. Tenascin-C expression was significantly lower in the SHCC cases and its distribution differed between SHCC and ICC. Matrix metalloproteinase-7 (MMP 7) expression was significantly higher in SHCC compared with HCC. Almost all stromal cells were alpha-smooth muscle actin-positive both in SHCC and ICC, whereas glial fibrillary acid protein (GFAP)-positive stromal cells were significantly more increased in ICC than in SHCC. SHCC clearly differed from HCC with respect to collagen types I, III and MMP-7 expression, and from ICC with regard to stromal components including laminin-5, tenascin-C and GFAP(+) stromal cells. PMID- 16271084 TI - CD117 expression in diffuse large B-cell lymphomas: fact or fiction? AB - CD117 (KIT) is expressed in a variety of hematopoietic neoplasms but there are a paucity of data regarding its expression in diffuse large B-cell lymphomas (DLBCL). The purpose of the present paper was to describe the authors' experience of two CD117+ DLBCL (one of follicle center-cell origin and one nasal Epstein Barr virus (EBV)- plasmablastic lymphoma associated with lytic bone lesions), as determined by tissue immunohistochemistry and flow cytometry. The CD117 expression in DLBCL was further evaluated using tissue microarrays and seven additional plasmablastic lymphomas, using two commercially available anti-CD117 antibodies (Ab-1, Oncogene and A4502, DakoCytomation). Membranous +/- cytoplasmic staining was seen with Ab-1 in 24/65 (37%) DLBCL, including 21/56 microarray DLBCL, two index cases, and 1/7 additional plasmablastic lymphomas, with persistent staining in 13% of microarray DLBCL despite preincubation with KIT peptide. However, A4502 had only membranous staining of the index cases and one additional EBV- plasmablastic lymphoma with medullary disease. The present study suggests that (i) CD117 expression can be detected sporadically in DLBCL of follicle center-cell origin and a subset of plasmablastic lymphomas; (ii) staining for CD117 might help in identifying EBV- plasmablastic lymphomas associated with bone marrow involvement; and (iii) CD117 antibodies should be carefully validated prior to use, because non-specific staining, as observed with Ab-1, could lead to false-positive results. PMID- 16271086 TI - New genetic model rat for congenital cataracts due to a connexin 46 (Gja3 ) mutation. AB - A rat strain with congenital nuclear cataracts has been established. Segregation analyses indicated that this phenotype had an autosomal recessive mode of inheritance, implying that a loss of function mutation of a single autosomal gene was responsible. The gene was mapped to the D15Rat6 locus on chromosome 15 through a linkage analysis using 93 backcrossed rats. The connexin 46 gene (Gja3) was found to be located close to the locus, and was regarded as a strong candidate because of its pivotal role in the lens fiber cells. Expression of the gene in the lens was comparable between the cataract and control rats when evaluated with immunohistochemistry and reverse transcription-polymerase chain reaction. However, a non-conservative missense mutation, Glu42Lys, was found in the gene of the cataract rats, which was likely to be responsible for the pathogenesis. This strain will be useful in pathophysiological studies on nuclear cataracts. PMID- 16271087 TI - Hepatocellular carcinoma with mesothelioma-like dissemination. AB - Reported herein is a case of hepatocellular carcinoma (HCC) with unusual peritoneal dissemination masquerading as peritoneal mesothelioma. A 61-year-old man was clinically found to have multiple tumors in his abdominal cavity; peritonitis carcinomatosa was suspected. An autopsy revealed numerous tumors of various sizes in the abdominal serosa, omentum, and diaphragm. No signs of tumor, fibrosis, or cirrhosis were found in the liver, except for a small nodule in the hepatic triangular ligament. Histologically, the tumor cells proliferated in thick trabeculae or in sheets and formed a few canaliculi and tubules with homogenously brown contents in their lumina, which stained positively with Hall stain. Immunohistochemically, these tumors were positive for hepatocyte, alpha fetoprotein (AFP) and low-molecular-weight cytokeratin; were focally positive for pan-cytokeratin and epithelial membrane antigen (EMA); and were negative for high molecular-weight cytokeratin, vimentin, and calretinin. Carcinoembryonic antigen (CEA) produced a bile canalicular immunohistochemical staining pattern. Thus, the tumor was diagnosed as an HCC (Edmondson II type) of the triangular ligament with massive peritoneal dissemination. The origin of this tumor and its differential diagnosis (malignant mesothelioma, hepatoid adenocarcinoma, and hepatoid yolk sac tumor) are discussed. PMID- 16271088 TI - Pituicytoma incidentally found at autopsy. AB - Pituicytoma is a rare benign neoplasm, occurring in the sellar and suprasellar regions. Reported herein is a case of asymptomatic pituicytoma, discovered at autopsy, in a 54-year-old Japanese woman. This is the first case report of pituicytoma, found incidentally at autopsy (incidentaloma), in which whole mounted sections are available for histological and immunohistochemical studies. Grossly, the bisected pituitary gland revealed a round, white to light tan, 7 mm diameter nodule. Microscopically, whole-mounted sections revealed a well circumscribed nodule with no fibrous capsule, located mainly in the neurohypophysis and partially compressing the adenohypophysis. The tumor was composed primarily of bipolar, occasionally unipolar, cells with syncytial fibrillary cytoplasm, arranged in short curvilinear fascicles and/or storiform patterns. Unusual histological features were seen, which included a few groups of large pleomorphic tumor cells with abundant, glassy, eosinophilic cytoplasm, occasionally associated with multinucleated giant tumor cells, and scattered Herring bodies within the tumor. Immunohistochemically, the tumor showed diffuse strong expression of glial fibrillary acidic protein, S-100 protein, and vimentin. Epithelial membrane antigen immunoreactivity was focally observed, mainly in the large tumor cells. Distinction from other intrasellar tumors (granular cell tumor and pilocytic astrocytoma) is important. Because the immunohistochemical profiles of these tumors are similar, histological findings are crucial for distinction. PMID- 16271089 TI - Sebaceous metaplasia of the nasal glands. PMID- 16271090 TI - A patient with mixed connective tissue disease and mixed-type autoimmune hemolytic anemia. PMID- 16271091 TI - Glucocorticoid stimulation of neutrophil donors: a medical, scientific, and ethical dilemma. PMID- 16271092 TI - TRANSFUSION's public access policy. PMID- 16271093 TI - The risk of posterior subcapsular cataracts in granulocyte donors. AB - BACKGROUND: Therapeutic use of adrenal corticosteroids is a risk factor for the development of posterior subcapsular cataract (PSC). Because corticosteroids are given to donors of apheresis granulocytes (PMNs) to improve yield, this study was performed to determine the prevalence of PSCs in PMN donors relative to a matched control group of apheresis platelet (PLT) donors. STUDY DESIGN AND METHODS: This study was a cross-sectional study stratified by age, sex, and lifetime apheresis experience at three sites. Individuals who had made at least five PMN donations preceded by corticosteroids were eligible. The presence of PSC was ascertained by grading digital retroillumination images of both lenses. A random subset of participants underwent clinical eye examinations by ophthalmologists masked as to study group. A logistic regression model was used to compute odds ratios (ORs). RESULTS: Granulocyte donors had given a mean of 13 donations (range, 5-39 donations) over a mean period of 8.5 years (range, 0.3-25.2 years). The mean corticosteroid exposure, in cortisol equivalents, was 2840 mg (range, 1067-9040 mg). Six of 89 PMN donors had photographic evidence of PSCs versus 4 of 89 controls. This difference was not significant (OR, 1.54; 95% confidence interval [CI], 0.46-5.08). Five of 33 PMN donors and 3 of 30 PLT donors had evidence of PSC by clinical examination. This difference was also not significant (OR, 1.61; 95% CI, 0.35-7.39). CONCLUSION: This study does not support the hypothesis that corticosteroid stimulation of PMN donors is associated with an increased risk of developing a PSC. PMID- 16271094 TI - Prevalence, incidence, and residual risk of human immunodeficiency virus among community and replacement first-time blood donors in Sao Paulo, Brazil. AB - BACKGROUND: Concerted efforts have been directed toward recruitment of community rather than replacement donors in Brazil. Time trends and demographic correlates of human immunodeficiency (HIV) prevalence and incidence among first-time (FT) donors in Brazil were examined by donation type. HIV residual risk from FT-donor transfusions, and projected yield of p24 antigen and nucleic acid test (NAT) screening were estimated. STUDY DESIGN AND METHODS: HIV prevalence data and seroreactive specimens were obtained at Fundacao Pro-Sangue/Hemocentro-de-Sao Paulo from 1995 to 2001. To estimate incidence, confirmed-positive samples from July 1998 through December 2001 were tested with a less-sensitive (detuned) enzyme immunoassay to detect recent seroconversions. Incidence data were used to estimate residual risk and p24 and NAT yield based on published window periods (WPs). RESULTS: HIV prevalence was 22 percent higher among the FT community donors than replacement donors (19.6 vs. 16.1 per 10,000; p < 0.01) and 48 percent higher among men than women (19.1 vs. 12.9; p < 0.01). In the multivariable logistic regression, both variables remained significant predictors of HIV prevalence. HIV prevalence decreased from 20.4 (1995) to 13.1 per 10,000 FT donations (2001). HIV incidence was 2.7 per 10,000 person-years. The estimated rate of infected antibody-negative donations was 14.9 per 1,000,000 units (95% confidence interval, 9.8-20.0). It was estimated that addition of p24 antigen, minipool NAT, and individual-donation NAT assays would detect 3.9 (2.0-5.8), 8.3 (5.3-11.3), and 10.8 (7.1-14.5) WP units per 1,000,000 FT donations, respectively. CONCLUSION: HIV incidence and residual transfusion risk estimates are approximately 10 times higher in Brazil FT donors compared to US and European FT donors. Community FT donors had higher HIV prevalence than replacement FT donors. The yield of p24 antigen or RNA screening will be low in Brazilian donors, but substantially higher than in US donors. PMID- 16271095 TI - Estimating the probability of a blood donation adverse event based on 1000 interviewed whole-blood donors. AB - BACKGROUND: Estimating the probability that a donor will have or not have an adverse event is useful for staff knowledge to give blood donors reassurance upon request. STUDY DESIGN AND METHODS: One-thousand donors from the general donor pool were interviewed for seven potential adverse events 3 weeks after a 525-mL whole-blood phlebotomy. The four most common adverse events were bruise (22.7%), sore arm (10.0%), fatigue (7.8%), and donor reaction (7.0%). A stepwise logistic regression analysis was performed based on five donor characteristics that were studied: age, weight, sex, race, and first-time donor status. The contribution of each significant or marginally significant factor to each adverse event was quantified. RESULTS: For donor reaction, weight (p < 0.0001) and age (p = 0.015) were significant contributors, and first-time donor status (p = 0.054) was a marginally significant contributor. An equation was derived, and the donor reaction rate can be estimated for a group based on the donor's weight, age, and first-time donor status. Similar analyses were performed for fatigue, sore arm, and bruise. CONCLUSION: Based on the derived formulas and with the use of a spreadsheet, data can be entered and the probability that a donor will have (or not have) a donor reaction, fatigue, sore arm, or bruise can be estimated. PMID- 16271096 TI - Comparison of cytomegalovirus polymerase chain reaction and serology for screening umbilical cord blood components. AB - BACKGROUND: Recipients of umbilical cord blood (UCB) transplants are susceptible to opportunistic infections, including cytomegalovirus (CMV). To prevent CMV transmission from UCB donors, most laboratories perform serology on corresponding maternal samples and quarantine units when the mother has immunoglobulin M (IgM) anti-CMV. STUDY DESIGN AND METHODS: UCB units and associated samples (UCB plasma and red cell pellet; maternal whole blood and serum) from two cord blood banks were tested with two validated CMV polymerase chain reaction assays (UL54 and UL93 targets). Results were compared with maternal CMV serology (IgG and IgM). RESULTS: Only 4 of 48 (8.3%) quarantined CMV IgM-positive units were also CMV nucleic acid testing (NAT)-positive (651-68,600 copies/mL). In contrast, 1 of 200 "CMV-safe" UCB units (CMV IgM-equivocal or -negative) had CMV DNA (0.5%). The corresponding maternal samples were CMV NAT-negative. Positive maternal IgM serology demonstrates only modest sensitivity (80%) and specificity (82%) and poor positive predictive value (8%), when correlated with the presence of CMV DNA in UCB units. CONCLUSION: CMV NAT may be a useful adjunct to serologic screening, potentially reducing wastage of IgM-positive and NAT-negative units while also detecting potentially infectious units that would pass serologic screening. A prospective clinical trial to further evaluate the role of CMV NAT in UCB transplantation appears warranted. PMID- 16271097 TI - Peripheral blood progenitor cell mobilization and collection in 42 patients with primary systemic amyloidosis. AB - BACKGROUND: High-dose chemotherapy followed by an inoculum of autologous peripheral blood progenitor cells (PBPCs) can improve survival in patients affected with primary systemic amyloidosis (AL). It has been documented, however, that the morbidity and mortality of PBPC mobilization and collection in this setting are higher than in patients with other diseases. To minimize the mobilization and collection-related risks, we developed a multidisciplinary approach involving different specialists to manage AL patients with predominant heart and renal involvement. STUDY DESIGN AND METHODS: We report our experience in 42 patients (23 men, 19 women; median age, 51.2 years; range, 28-68 years) with AL who underwent PBPC mobilization and collection. Twenty of the 42 patients (47.6%) had cardiac involvement and 35 of 42 (83.3%) renal involvement. Thirty three patients (78.5%) were mobilized with granulocyte-colony-stimulating factor (G-CSF) alone (10 microg/kg) and 9 (21.4%) with cyclophosphamide (CTX) (3 g/m(2)) plus G-CSF (10 microg/kg). RESULTS: The median number of collections per patient after either G-CSF or CTX plus G-CSF was 1.8 (range, 1-3). The median number of CD34+ cells collected in patients mobilized with G-CSF alone was 8.2 x 10(6) per kg (range, 1.35 x 10(6)-21.3 x 10(6)/kg) and in patients mobilized with CTX plus G-CSF it was 8.9 x 10(6) per kg (range, 5.5 x 10(6)-14.9 x 10(6)/kg). Forty of the 42 (95.2%) patients produced the minimum required CD34+ cell target dose (4 x 10(6)/kg). The overall rate of morbidity during the collections was 50 percent (21/42 patients): 18 patients (42.8%) had asymptomatic hypotension, 1 (2.4%) had symptomatic hypotension with nausea and vomiting, and 2 (4.7%) experienced a life threatening hypotensive episode. There were no procedure-related deaths. CONCLUSION: Our multidisciplinary approach was effective in limiting the serious side effects related to PBPC mobilization and collection in AL patients. PMID- 16271098 TI - Long-term transfusion of polymerized bovine hemoglobin in a Jehovah's Witness following chemotherapy for myeloid leukemia: a case report. AB - A 52-year-old female Jehovah's Witness presented with relapsed secondary acute myeloid leukemia. Because of chemotherapy-induced anemia, she was infused with the bovine hemoglobin (Hb)-based oxygen carrier HBOC-201 (Biopure) as the sole means of transfusion support. HBOC-201 has only been used for management of acute hemorrhage, and its utility in providing longer term transfusion support is unknown. Over a period of 18 days, a total dose of 1230 g of HBOC-201 was delivered. Although the patient succumbed to the disease after 18 days of treatment, this case documents our experience with the highest dose and duration of HBOC-201 ever used. Although possible renal toxicity could not be definitively excluded, the homogeneous extraction of oxygen by the brain in the presence of and perhaps from HBOC-201 was demonstrated. PMID- 16271100 TI - Platelet glycolytic flux increases stimulated by ultraviolet-induced stress is not the direct cause of platelet morphology and activation changes: possible implications for the role of glucose in platelet storage. AB - BACKGROUND: Stress-enhanced platelet (PLT) storage lesions include increased glycolysis, discoid-to-sphere morphology change, and spontaneous PLT activation. It is not clear if reduction in glycolysis can alleviate storage lesion development. STUDY DESIGN AND METHODS: Apheresis PLT concentrates were exposed to 17.2 J/mL UV light and 50 microM riboflavin, followed by storage with various concentrations of 2-deoxyglucose (2-DOG) for 5 days. The control had no UV or 2 DOG exposure. RESULTS: Lactate production and glucose consumption were increased significantly to 0.1371 +/- 0.0281 and 0.0724 +/- 0.0151 mmol per 10(12) cells per hour for UV-treated PLTs, respectively, when compared to control samples. UV treatment induced a decline in pH to 6.55 +/- 0.26 for treated PLTs on Day 5, hypotonic shock response (HSR) 33 +/- 25 percent, extent of shape change (ESC) to 3.8 +/- 3.6 percent, swirl 1.0 +/- 1.0 and increased P-selectin expression 85.2 +/- 9.4 percent. Addition of 2-DOG up to 20 mmol per L significantly reduced lactate production to 0.0515 +/- 0.0045 mmol per 10(12) cells per hour (p < 0.05) and glucose consumption to 0.0293 +/- 0.0060 mmol per 10(12) cells per hour and increased pH to 7.35 +/- 0.09 in a dose-dependent manner. 2-DOG, however, had no effects on HSR, ESC, swirl, and P-selectin expression. Furthermore, an exaggeration of UV-stressed PLT aggregation by addition of 2-DOG was also observed. CONCLUSIONS: Increased glycolytic flux is not a direct cause for PLT morphology change and spontaneous activation during storage lesion development. Reduction of glucose utilization may increase PLT loss during storage. PMID- 16271099 TI - Therapeutic efficacy and safety of red blood cells treated with a chemical process (S-303) for pathogen inactivation: a Phase III clinical trial in cardiac surgery patients. AB - BACKGROUND: A randomized, double-blind trial is reported of the clinical efficacy of red blood cells (RBCs) treated for pathogen inactivation with S-303, a synthetic labile alkylating agent. STUDY DESIGN AND METHODS: Patients undergoing complex cardiac surgeries were randomly assigned to receive either S-303-treated (test) or conventional (control) RBC transfusion during surgery and for 6 days thereafter. Efficacy was evaluated by comparing the occurrence of a composite primary endpoint of treatment-related morbidity (myocardial infarction and renal failure) and mortality. RESULTS: Two-hundred twenty-three patients were randomly assigned and 148 patients who received transfusions (74 with S-303-treated RBCs and 74 with control RBCs) were evaluable. The incidence of the primary endpoint was equivalent between the two groups (22 and 21% in the S-303-treated and control RBC groups, respectively). Secondary endpoints, including hemoglobin increment (mean, 1.4 vs. 1.5 g/dL), number of RBC transfusions (mean, 4.4 vs. 3.8 units), and other blood product support, were also comparable. The adverse event profile was similar between groups; however, patients who received S-303 RBCs were significantly more likely to develop constipation and less likely to suffer supraventricular extrasystoles. Four patients (2 test and 2 control) demonstrated positive indirect antiglobulin tests with reactivity for S-303 RBCs at one or more time points before or after transfusion, without evidence of hemolysis. CONCLUSION: S-303-treated and conventional RBCs were equivalent with respect to clinical efficacy and safety in supporting the transfusion needs of cardiac surgery patients. Investigations are under way to ascertain the significance of S 303 RBC antibodies and to prevent their occurrence. PMID- 16271101 TI - Tissue regeneration and in loco administration of platelet derivatives: clinical outcome, heterogeneous products, and heterogeneity of the effector mechanisms. AB - BACKGROUND: In loco administration of platelet (PLT) derivatives is a relatively new auxiliary treatment for tissue regeneration to be hastened. Enthusiastic reports are faced by more critical ones. The more obvious rationale for the in vivo administration of PLT derivatives resides in their growth factor content. STUDY DESIGN AND METHODS: The relevant literature was systematically reviewed. Close scrutiny of the technical details was carried out to find out the procedural differences accounting for conflicting results. RESULTS: An impressively vast heterogeneity of conduct was found in both in vitro and in vivo studies. Major outcome-affecting variables were recognized such as those associated with PLT preparation; growth factor measurement; proliferation test; dose, timing, and administration of the PLT derivatives; study design; and primary endpoints. CONCLUSIONS: So many variables were found making standardization or confrontation of the in vitro and the in vivo studies barely conceivable or manageable. The mechanisms of action are very complex. The attribution of tissue regeneration capacity of PLT derivatives solely to the PLT derived growth factors is simplistic. The results obtained through in vitro experiments are indicative for general mechanisms. Their simplistic hold to the complex in vivo environment may be misleading. PMID- 16271102 TI - Bicarbonate inhibits the growth of Staphylococcus epidermidis in platelet concentrates by lowering the level of non-transferrin-bound iron. AB - BACKGROUND: Platelet concentrates (PCs) contain non-transferrin-bound iron (NTBI) owing to the displacement of iron from plasma-derived transferrin by citrate. NTBI in the PC medium supports the growth of Staphylococcus epidermidis. The possibilities of lowering the level of NTBI have been studied with the aim to inhibit the growth of S. epidermidis in the PC medium. STUDY DESIGN AND METHODS: NTBI in PC supernatants was determined by a chelation method and by the bleomycin detectable iron assay. Iron binding by transferrin was determined by spectrophotometry. The growth of inoculated S. epidermidis in PC supernatants was monitored by optical density and determination of viable counts. RESULTS: Bicarbonate enhanced in a dose-dependent manner transferrin iron binding in citrate-containing solutions, including citrated plasma and PAS-II. The use of a modified anticoagulant supplemented with bicarbonate effectively lowered the level of NTBI and inhibited bacterial growth in citrated plasma. Supplementation of bicarbonate to the additive solution to increase the ratio of bicarbonate to citrate in a reconstituted PC medium further inhibited bacterial growth. Maintenance of stable pH and bicarbonate level in the reconstituted medium necessitated storage under 5 percent CO(2). CONCLUSIONS: The relatively low bicarbonate level in PC medium promotes iron displacement by citrate from plasma derived transferrin. The appearance of NTBI can be decreased and iron-dependent bacterial growth can be inhibited by increasing bicarbonate level in citrated plasma and PC medium. To achieve the same beneficial effect in blood banking, other more practical ways to bind NTBI in a harmless form should be developed. PMID- 16271104 TI - Beneficial effect of transfusion with low-affinity red blood cells in endotoxemia. AB - BACKGROUND: Sepsis caused by endotoxins such as lipopolysaccharide (LPS) impairs the microcirculation, diminishing tissue blood supply and aggravates systemic hypoxia. A novel lower-affinity hemoglobin (Hb) variant, Hb Presbyterian, enhances oxygen release to peripheral tissues and may improve tissue oxygen supply during sepsis. STUDY DESIGN AND METHODS: This study investigated the effectiveness of Presbyterian Hb in transfusion therapy with LPS-challenged sepsis mouse model. Septic wild-type mice were transfused with RBCs from Presbyterian Hb-carrying mutant mice and wild-type mice. Their survival rates were assessed, and apoptosis of hepatocytes was evaluated. Survival rates of septic Presbyterian mutant mice and the wild-type littermates were also studied. RESULTS: The Presbyterian mutant RBC-transfused septic group survived longer than the wild-type RBC-transfused group. Apoptosis was reduced in the hepatocytes of the former group. Presbyterian mutant mice themselves, however, did not have stronger resistance to LPS-induced sepsis. CONCLUSION: Transfusion of low affinity Hb-containing RBCs has beneficial effects in septic mice. PMID- 16271103 TI - Treatment with C1 inhibitor concentrate in abdominal pain attacks of patients with hereditary angioedema. AB - BACKGROUND: Abdominal edema attacks in patients with hereditary angioedema are often extremely painful, associated with vomiting and diarrhea, and have a high potential for causing recurrent disability of the patient. STUDY DESIGN AND METHODS: Intraindividual comparison of retrospective data in 75 hereditary angioedema patients comprising 4,834 abdominal attacks treated with C1 inhibitor concentrate versus 17,444 untreated abdominal attacks. RESULTS: The mean duration of abdominal attacks was 92.0 hours (SD, 40.8 hr) when untreated compared to 39.9 hours (SD, 30.0 hr) when treated. Patients reported a mean maximal pain score of 8.6 (SD, 1.7; range, 1-10) for untreated attacks compared to 4.5 (SD, 2.9) when treated. Vomiting occurred in 83.3 percent of untreated attacks and in 6.0 percent of treated attacks, respectively. Diarrhea was reported in 41.8 percent of untreated attacks and in 11.0 percent of treated attacks, whereas cardiovascular collapse due to hypovolemia was observed in 3.5 percent of untreated attacks versus 0.1 percent in treated attacks. Mean time to relief of symptoms was 53.5 minutes when treated early compared to 114 minutes when treatment was delayed. No drug-related adverse or serious adverse events were observed as far as the injections were performed correctly. CONCLUSION: C1 inhibitor concentrate is highly effective and safe in treating abdominal attacks in patients suffering from hereditary angioedema. PMID- 16271105 TI - Serial blood donations for intrauterine transfusions of severe hemolytic disease of the newborn with the use of recombinant erythropoietin in a pregnant woman alloimmunized with anti-Ku. AB - BACKGROUND: The management of a pregnant woman with the rare Ko phenotype and anti-Ku is a special challenge, because matched blood is extremely rare and the possibility of severe hemolytic disease of the newborn is high. CASE REPORT: A 30 year-old woman with rare Ko (Knull) phenotype presented at 18 weeks of gestation with positive indirect agglutination test results. She had anti-Ku due to previous blood transfusion, one pregnancy, and two abortions. STUDY DESIGN AND METHODS: During this pregnancy, anti-Ku titers ranged from 1024 to 4096. At the 26th week of gestation ultrasound showed a hydropic fetus and urgent intrauterine exchange transfusion was performed with the maternal red blood cells (RBCs). Recombinant human erythropoietin (rHu-EPO) and intravenous (IV) iron were administered to the mother to ensure an adequate supply of matched RBCs for intrauterine transfusions and possible perinatal hemorrhage. RESULTS: Intrauterine transfusions were repeated every 1 to 3 weeks. By 35 weeks 2 days of gestation, the mother had donated 4 units of blood, and four intrauterine transfusions had been performed. Cesarean section was then decided and a healthy male newborn was born. He was treated with phototherapy but without exchange transfusions. By the 15th day of life rHu-EPO was administrated to the newborn because of anemia. The maternal RBCs completely disappeared from the child's blood by Day 100. CONCLUSIONS: As shown in this case, treatment with rHu-EPO and IV Fe has effectively increased the mother's capacity to donate RBCs for autologous use and intrauterine transfusions, with no adverse effects to the mother or the child. PMID- 16271106 TI - Rhnull syndrome: identification of a novel mutation in RHce. AB - BACKGROUND: The deficiency of Rh proteins on red blood cells (RBCs) from individuals of the Rh(null) amorph type are the result of homozygosity for a silent RHCE in cis with a deleted RHD. A novel mutation in RHce was identified in two Caucasian Brazilian girls with the amorph type of Rh(null) who were born to parents who were first cousins. STUDY DESIGN AND METHODS: RBCs from the Rh(null) sisters and from family members were analyzed by serology and flow cytometry with specific antibodies. Genomic DNA and transcripts were tested by polymerase chain reaction and sequence analysis. RESULTS: Rh(null) RBCs were nonreactive with anti Rh and anti-LW. Molecular analyses showed a deletion of RHD and of one nucleotide (960/963; GGGG-->GGG) in exon 7 of the RHce. This deletion introduced a frameshift after Gly321, a new C-terminal sequence, and a premature stop codon, resulting in a shorter predicted protein with 357 amino acids. CONCLUSION: The detection of a unique RHce transcript indicated that the two sisters were homozygous, whereas the other family members were heterozygous for the mutation. A novel mutation resulting in the amorph Rh(null) with loss of Rh antigen expression is described. PMID- 16271107 TI - Anti-HPA-9bw (Maxa) fetomaternal alloimmunization, a clinically severe neonatal thrombocytopenia: difficulties in diagnosis and therapy and report on eight families. AB - BACKGROUND: Fetal or neonatal alloimmune thrombocytopenia (FMAIT) results from a maternal alloimmunization against fetal platelet (PLT) antigens. In Caucasian persons, HPA-1a is the most frequently implicated antigen. During the past few years, FMAIT has been reported associated with rare or private antigens. STUDY DESIGN AND METHODS: Since the first documented case of FMAIT due to anti-HPA-9bw (Max(a)), no additional cases have been reported. Here a retrospective analysis is presented of the cases referred to our laboratories in recent years. The diagnosis was performed by genotyping and identification of the maternal alloantibody by the monoclonal antibody-specific immobilization of PLT antigens (MAIPA) technique. RESULTS: Parental genotyping showed HPA-9bw (Max(a)) mismatch as the sole antigenic incompatibility in seven of eight families. Because the father was found to be HPA-9bw (Max(a)) heterozygous in all the cases, the infant or fetus was genotyped to ascertain the diagnosis. The maternal alloantibody was identified in the MAIPA technique. These data strongly suggest, however, that recognition of the HPA-9bw (Max(a)) epitope is not uniform. The neonatal thrombocytopenia was severe in most cases with bleeding. The outcome was good in all the cases but one. CONCLUSION: This analysis confirms that anti-HPA-9bw (Max(a)) FMAIT is not uncommon and was found to be approximately 2 percent of our confirmed FMAIT cases. It is a clinically severe syndrome that requires prompt diagnosis, albeit difficult, and maternal PLT transfusion therapy. Laboratory investigation of a suspected FMAIT case should be carried out in a specialist laboratory well-experienced in optimal testing. Appropriate management and antenatal therapy should be considered for successive pregnancies to prevent fetal bleeding. PMID- 16271108 TI - Demonstrable parasitemia among Connecticut blood donors with antibodies to Babesia microti. AB - BACKGROUND: Reports of transfusion-transmitted Babesia microti have risen steadily during the past several years, reflecting a concurrent increase in US cases of human babesiosis. Although several studies have measured B. microti antibodies in blood donors, little is known about associated parasitemia and the inherent risk of transmitting the parasite by transfusion. STUDY DESIGN AND METHODS: Donations from blood donors located in Babesia-endemic and nonendemic areas of Connecticut were tested for B. microti antibodies from July through September. Subsequently, an additional blood sample was collected from selected seropositive donors and tested by nested polymerase chain reaction (PCR) for B. microti nucleic acids. RESULTS: A total of 3490 donations, 1745 each from endemic and nonendemic areas, were tested for B. microti antibodies; 30 (0.9%) were confirmed as positive and seroprevalence rates peaked in July. Significantly more seropositive donations were from endemic areas (24, 1.4%) than nonendemic areas (6, 0.3%). Ten (53%) of 19 seropositive donors subsequently tested by PCR were positive. CONCLUSION: B. microti seroprevalence was highest in those areas of Connecticut where the parasite is endemic. More than half of seropositive donors tested had demonstrable parasitemia, indicating that many are at risk for transmitting B. microti by blood transfusion. Three donors were identified as parasitemic in October, suggesting that donors may be at risk for transmitting the parasite outside of the peak period of community-acquired infection. PMID- 16271109 TI - Exposure of hematologic patients to parvovirus B19 as a contaminant of blood cell preparations and blood products. AB - BACKGROUND: Patients with hematologic malignancies often require blood products, and parvovirus B19 is known to be transmitted by this route. Primary infection with parvovirus B19 shows a wide variety of disease manifestation. In immunocompromised patients, symptoms are severe and viral clearance is delayed or missing. STUDY DESIGN AND METHODS: A total of 2123 blood products given to all patients of a hematologic ward over a period of 6 months were retrospectively examined for the presence of parvovirus B19 DNA by an in-house real-time polymerase chain reaction (PCR; TaqMan). Patients who had received B19 DNA positive blood products were further investigated serologically and by PCR for the presence of parvovirus B19 antibodies and DNA. RESULTS: Twenty-one (1%) of 2123 blood products tested positive for the presence of B19 DNA (2% of pooled products, 0.7% of single-donor products, and 17.6% of allogeneic peripheral blood progenitor cells), the median viral load was 700 genome equivalents per mL. During the study period, 114 patients were treated on the ward, and 14 (12%) of them received B19 DNA-positive blood components. None of them developed symptoms of an acute B19 infection, although one had a short low-level viremia. CONCLUSIONS: Although B19 DNA was detected in 1 percent of blood products given to hematologic patients, the exposure of 12 percent of patients did not result in symptomatic infections. PMID- 16271110 TI - Significance of the signal-to-cutoff ratios of anti-hepatitis C virus enzyme immunoassays in screening of Chinese blood donors. AB - BACKGROUND: The correlation between signal-to-cutoff (S/CO) ratios of a second generation hepatitis C virus (HCV) enzyme immunoassay (EIA; Abbott) and a third generation HCV enzyme-linked immunosorbent assay (ELISA; Ortho) and confirmed HCV infection has been reported. The utility of the values for the Chinese anti-HCV EIA kits, however, has not been studied in evaluating test results in Chinese blood donors. STUDY DESIGN AND METHODS: A total of 156 donor samples repeat reactive for anti-HCV at routine screening from five representative regions of China were retested for anti-HCV by the Ortho third-generation HCV ELISA and six Chinese EIA kits and for HCV RNA by a human immunodeficiency virus-1 and HCV assay (Procleix, Chiron Corp.). The HCV RNA-nonreactive samples were further tested for anti-HCV by a third-generation recombinant immunoblot assay RIBA (Chiron Corp.). The positive result by either nucleic acid amplification test or RIBA was interpreted as confirmed HCV infection. RESULTS: The confirmed HCV prevalence rate in donors in five representative regions obtained in this study was 0.20 percent (77/37,900) in 2004. All seven anti-HCV EIA kits had a significant correlation between S/CO ratios and confirmed HCV infection. The threshold S/CO ratios, which predicted more than 95 percent of confirmed HCV infections for the Ortho, SABC, BGI-GBI, InTec, GWK, KHB, and WANTAI kits, were 3.8, 6.0, 7.0, 8.6, 10.0, 10.0, and 14.0, respectively. CONCLUSIONS: Anti-HCV EIA kits commonly used in Chinese donors screening demonstrate good correlation between S/CO ratios and the confirmed infection. For the Ortho third-generation HCV ELISA, the S/CO ratio of 3.8 determined by the US Centers for Disease Control and Prevention is applicable to Chinese blood donors. The Chinese domestic EIA kits evaluated show a diverse range of threshold S/CO ratios. PMID- 16271111 TI - J.J. (Joghem) van Loghem. PMID- 16271112 TI - Tibor Jack Greenwalt. PMID- 16271114 TI - Newborns' sex and hematopoietic progenitor cell content of cord blood. PMID- 16271115 TI - The effect of storage of fresh-frozen plasma at -80 degrees C for as long as 14 years on plasma clotting proteins. PMID- 16271117 TI - A call for a return to patient-centered care. PMID- 16271118 TI - Sphere of Nursing Advocacy Model. AB - The Sphere of Nursing Advocacy (SNA) model explains and depicts nursing advocacy on behalf of a client. The SNA model views the client as continually protected from the external environment by a semipermeable sphere of nursing advocacy that allows clients to self advocate if the client is emotionally and physically able or to be advocated for by the nurse if the patient is unable to advocate for him- or herself. The SNA model can be used to guide research or it can provide the basis for instruction on the subject of nursing advocacy. PMID- 16271120 TI - The nursing shortage: can we look to teachers as a source of support? PMID- 16271119 TI - Sedentarism: a concept analysis. AB - TOPIC: Concept analysis of sedentarism. PURPOSE: To analyze the concept of sedentarism and provide a definition and model of sedentarism to guide practice and research. SOURCES: Published literature. CONCLUSIONS: Prevention and treatment of sedentarism is a priority for healthcare providers serving all developmental groups. Research is urgently needed to describe the prevalence, risk factors, and consequences of sedentarism, and to identify the most effective intervention strategies and public policy changes to promote a physically active lifestyle. PMID- 16271121 TI - Health/Illness transition and telehealth: a concept analysis using the evolutionary method. AB - TOPIC: Telehealth as an alternative to hospitalization. PURPOSE: Exploring the health/illness transition that occurs when an acutely ill client may be discharged into the home with telehealth in place of hospitalization or in an effort to promote an earlier discharge from the hospital. SOURCE: Published literature. CONCLUSION: The concept of the health/illness transition using telehealth has not been researched. Qualitative methods of research using interviews could enhance the knowledge in selecting appropriate individuals and nursing interventions to enhance the clients' transition from the hospital to the home with telehealth. PMID- 16271124 TI - One practice week at a glance. Today's nurse: the patient's advocate. PMID- 16271122 TI - Travel counseling for the elderly traveler. AB - As the baby boomer's generation retirees, many will have the time and money to travel abroad to see the world's exotic wonders or visit family and friends. When the travelers are elderly, they are particularly vulnerable to the effects of travel. Healthcare professionals are responsible for counseling elders on travel health based on their medical history, destination, method of transportation, and exposure risks. Important areas of travel counseling include preparing for travel, air travel, safety, sun and heat, insect precautions, food and water precautions, and vaccinations. PMID- 16271123 TI - Said another way. My definition of nursing. AB - The definition of nursing has evolved since the introduction of the definition by the American Nurses Association in 1980. The society we nurses serve has changed significantly and members of the society demand quality health care and respect for their individual decisions regarding their health care. As a profession, we must keep our fingers on the pulse of society and be ready to respond to the changes of society's needs. Being diligent in defining our position in society is imperative for the preservation of the nursing profession. This paper supports the current definition of the ANA (2003) within the domains of the art, science, and practice of nursing. PMID- 16271126 TI - Examining the future of professional journals. PMID- 16271127 TI - Educating nurses for the 21st century. AB - The current and projected demand for nurses and nursing services worldwide, coupled with dramatic changes in the delivery of health care, require nurses with more knowledge, more education, and more skills. Issues facing nursing require a reconceptualization of the approach and expectations for nursing practice and for the educational models and processes that lead to reformed nursing practice. New initiatives such as the Clinical Nurse Leader and the Doctor of Nursing Practice may lead the transformational process that is needed. PMID- 16271128 TI - Effects of acculturation and social exchange on the expectations of filial piety among Hispanic/Latino parents of adult children. AB - This study explores the effects of acculturation and social exchange variables on the expectations of filial piety (values and beliefs about parents' care) among Hispanic/Latino parents of adult children. A convenience sample of 318 Hispanics/Latinos with adult children was assembled in Texas and North Carolina, USA, for face-to-face interviews in order to collect information about their families and to assess their expectations of filial piety by their adult children. The number of years in the USA, age, gender, the eldest child's age, and living with a person > or = 65 years of age had significant relationships to scores from a new Spanish version of the Expectations of Filial Piety Scale (EFPS). The significant variables explained 46.3% of the variance in the EFPS scores. A conceptual model that includes both acculturation and social exchange variables is a useful approach for understanding the expectations of filial piety by Hispanic/Latino parents of adult children. PMID- 16271129 TI - Nurses' attitudes and practices towards abused women. AB - The present article is a descriptive survey assessing nurses' knowledge, training, and practices regarding the care of abused women. A self-administered questionnaire was sent to all nurses working within the government health organization on the Aland Islands, Finland. The response rate was 57%. Most nurses had no formal training in domestic violence and were less likely than the in-service-trained nurses to intervene with abused women. Nurses' knowledge, beliefs, and practices were found to be unsystematic and had drawbacks. Nurses were not familiar with the formal structure provided by the health administration. The results suggest that in order to meet the urgent need for training among nurses, an in-service training program needs to be set up. This program would help the nurses in identifying survivors of abuse and make them prepared to intervene more effectively to promote the health of these women. PMID- 16271130 TI - Attitudes toward advance directives and the impact of prognostic information on the preference for cardiopulmonary resuscitation in medical inpatients in Chiang Mai University Hospital, Thailand. AB - Our previous study revealed that cardiopulmonary resuscitation (CPR) was performed in 65.7% of 411 terminally ill patients who died in a tertiary-care university hospital in northern Thailand. Advance directives (ADs) are needed to ensure that life-sustaining therapies are used more appropriately. To investigate inpatients' attitudes regarding ADs for CPR and the impact of providing prognostic information on treatment preferences for CPR, we interviewed a randomly selected group of 200 ambulatory medical inpatients in multiple sessions. The results showed that most subjects had a positive attitude towards ADs for CPR. The majority preferred to have CPR when no information was provided on the chance of survival. However, this proportion decreased depending on the prognostic scenarios. Our investigation suggested that the preference of patients for CPR should be assessed individually and gradually, with adequate information given on the chance of survival. PMID- 16271131 TI - What characterizes the 'usual' preoperative education in clinical contexts? AB - The literature on preoperative education is dominated by studies that employ experimental designs to measure the effects of structured programs on patient outcomes. These studies predominantly compare structured preoperative educational interventions with the usual care that patients receive. However, the notion of what the "usual" care comprises is largely elusive and unexplored. This study aimed to understand how the usual preoperative education is practiced in a number of surgical clinical units at one particular hospital in Ireland. Twelve experienced surgical nurses were interviewed in depth. A qualitative strategy resembling grounded theory was employed to analyze the data. The findings indicated that the content and quality of the preoperative education that patients received depended largely upon the individual nurse caring for the patient. Although there was a generic content of preoperative education that all participants identified as important, this related to physical and technical issues, and to the transmission of administrative or procedural information. Perspectives on the process of delivering preoperative education were diverse. The vast majority did not relate to the formal language of the discourses of teaching and learning; nonetheless, their descriptions of engaging in their work indicated the application of some elements of educational theory. The data suggested that the use of teaching tools was inconsistent, depending on their availability and the practices of individual nurses. On the whole, the usual care as described by participants in our study may be characterized as uneven, variable, and mutable. We conclude by raising some methodological issues relating to the use of the usual care in control groups in experimental studies on preoperative education. PMID- 16271132 TI - Menstrual status and bone mineral density among female athletes. AB - The present study investigated the relationship between menstrual status and bone mineral density (BMD). Sixty-three elite female athletes competing at the regional level participated. Self-reported menstrual status, stress during the past 6 months, dietary intake of calcium, blood samples for hormonal study, mid thigh skinfold thickness, triceps, iliac crest, spine and femoral neck BMD were determined. It was found that more than half of the athletes were eumenorrheic while almost half were menstrually dysfunctional. The bone mineral density at the lumbar spine and the femoral neck were within normal ranges. Menstrual dysfunction in female athletes was related to a low BMD at the lumbar spine but not at the femoral neck. Delayed menarche and menstrual dysfunction during the first 2 years after menarche were related to current menstrual dysfunction, but low percent body fat was not related to menstrual dysfunction. This study suggests that exercise in elite female athletes might be an underlying cause of menstrual dysfunction and that there is a relationship between lumbar spine BMD and menstrual dysfunction. The assessment of menstrual history and percent body fat could be used as a screening tool for menstrual dysfunction. PMID- 16271134 TI - Development of indicators to assess the quality of nursing care in Thailand. AB - In Thailand, quality of nursing care has been defined. The next step is to determine how to measure quality. Given the limited resources to provide health care and the demands to justify the use of registered nurses, there is an urgent need to demonstrate how professional nursing care makes a difference to outcomes of care. The purpose of this study was to further refine nursing indicators of quality by developing operational definitions, validating them and then determining their applicability in a variety of clinical settings. The process included three phases. The first phase used focus groups to identify and define indicators of quality nursing care and the second phase included testing these indicators in four settings to determine if data could be collected. Manuals were developed that defined the quality indicators and the scoring system to be used in the assessment. In the third phase, the findings were presented to a group of experts and minor changes were made to the indicators. Then, the indicators were categorized into three groups: structure, process, and outcome. The validated outcomes and manual for their assessment and monitoring were sent to all hospitals in Thailand. The study relied upon the findings from the four different hospitals. If the indicators were implemented in a wider variety of settings, other differences might have emerged. There is also a possibility that 'best practice' might not be reflective of the scientific basis of nursing practice. The findings generally support the initial work carried out in the USA. There is a need to continue to study how these indicators can improve patient outcomes. PMID- 16271133 TI - Implementing clinical guidelines for nutrition in a neurosurgical intensive care unit. AB - Patients in neurosurgical intensive care have individual needs concerning nutrition because of their conditions. An important therapeutic goal is to prevent the development of malnutrition as it contributes to an increase in mortality and enhances the risk of complications in these patients. The health care team has a mutual responsibility for this, although it is a complex task and a correct assessment of patients' nutritional needs is vital. Multidisciplinary clinical guidelines focused on nutrition might help the health-care staff in decision-making and allowing individualized treatment for patients. The aim of this study was to evaluate the implementation and use of such a guideline. The results show that the guideline is used in varying degrees by the health-care team. Areas that focused on more practical aspects of nutritional support seemed to have a unified approach, whereas areas on nutritional assessment and routine nutrition orders demonstrated both a lack of knowledge and unclear role responsibilities. The results reveal how different professional groups in health care perceive the implementation of a clinical guideline. Some areas need further clarification, there needs to be continuing development of nutritional guidelines, and education of staff is needed in order to enhance the nutritional care of patients. PMID- 16271135 TI - Critically appraising quantitative research. AB - Critical appraisal of quantitative research is a skill that is necessary for adequate evidence-based practice. This paper describes critical appraisal and offers a process for its implementation. Four categories of quantitative studies are reviewed and associated links to relevant critical appraisal frameworks are provided. The implications for students, nursing faculty members, and practicing nurses are discussed. PMID- 16271136 TI - The epidemiology of suicide and attempted suicide in Dutch General Practice 1983 2003. AB - BACKGROUND: Many patients attempting or committing suicide consult their general practitioner (GP) in the preceding period, indicating that GPs might play an important role in prevention. The aim of the present study was to analyse the epidemiology of suicidal behaviour in Dutch General Practice in order to find possible clues for prevention. METHOD: Description of trends in suicide and suicide attempts occurring from 1983-2003 in the Dutch General Practice Sentinel Network, representing 1% of the Dutch population. The data were analysed with regard to: 1) suicidal behaviour trends and their association with household situation; 2) presence of depression, treatment of depression and referral rate by GPs; 3) contact with GP before suicide or suicide attempt and discussion of suicidal ideation. RESULTS: Between 1983 and 2003 the annual number of suicide and suicide attempts decreased by 50%. Sixty percent of the patients who committed or attempted suicide were diagnosed as depressed, of whom 91% were treated by their GP with an antidepressant. Living alone was a risk factor for suicide (odds ratio 1.99; 95% CI 1.50 to 2.64), whereas living in a household of 3 or more persons was a relative risk for a suicide attempt (odds ratio 1.81; 95% CI 1.34 to 2.46). Referral to a psychiatrist or other mental health professionals occurred in 65% of the cases. GPs recalled having discussed suicidal ideation in only 7% of the cases, and in retrospect estimated that they had foreseen suicide or suicide attempts in 31% and 22% of the cases, respectively, if there had been contact in the preceding month. CONCLUSION: With regard to the prescription of antidepressants and referral of suicidal patients to a psychiatrist, Dutch GPs fulfil their role as gatekeeper satisfactorily. However, since few patients discuss their suicidal ideation with their GP, there is room for improvement. GPs should take the lead to make this subject debatable. It may improve early recognition of depressed patients at risk and accelerate their referral to mental health professionals. PMID- 16271138 TI - Why do snails have hairs? A Bayesian inference of character evolution. AB - BACKGROUND: Costly structures need to represent an adaptive advantage in order to be maintained over evolutionary times. Contrary to many other conspicuous shell ornamentations of gastropods, the haired shells of several Stylommatophoran land snails still lack a convincing adaptive explanation. In the present study, we analysed the correlation between the presence/absence of hairs and habitat conditions in the genus Trochulus in a Bayesian framework of character evolution. RESULTS: Haired shells appeared to be the ancestral character state, a feature most probably lost three times independently. These losses were correlated with a shift from humid to dry habitats, indicating an adaptive function of hairs in moist environments. It had been previously hypothesised that these costly protein structures of the outer shell layer facilitate the locomotion in moist habitats. Our experiments, on the contrary, showed an increased adherence of haired shells to wet surfaces. CONCLUSION: We propose the hypothesis that the possession of hairs facilitates the adherence of the snails to their herbaceous food plants during foraging when humidity levels are high. The absence of hairs in some Trochulus species could thus be explained as a loss of the potential adaptive function linked to habitat shifts. PMID- 16271137 TI - Cross-platform analysis of cancer microarray data improves gene expression based classification of phenotypes. AB - BACKGROUND: The extensive use of DNA microarray technology in the characterization of the cell transcriptome is leading to an ever increasing amount of microarray data from cancer studies. Although similar questions for the same type of cancer are addressed in these different studies, a comparative analysis of their results is hampered by the use of heterogeneous microarray platforms and analysis methods. RESULTS: In contrast to a meta-analysis approach where results of different studies are combined on an interpretative level, we investigate here how to directly integrate raw microarray data from different studies for the purpose of supervised classification analysis. We use median rank scores and quantile discretization to derive numerically comparable measures of gene expression from different platforms. These transformed data are then used for training of classifiers based on support vector machines. We apply this approach to six publicly available cancer microarray gene expression data sets, which consist of three pairs of studies, each examining the same type of cancer, i.e. breast cancer, prostate cancer or acute myeloid leukemia. For each pair, one study was performed by means of cDNA microarrays and the other by means of oligonucleotide microarrays. In each pair, high classification accuracies (> 85%) were achieved with training and testing on data instances randomly chosen from both data sets in a cross-validation analysis. To exemplify the potential of this cross-platform classification analysis, we use two leukemia microarray data sets to show that important genes with regard to the biology of leukemia are selected in an integrated analysis, which are missed in either single-set analysis. CONCLUSION: Cross-platform classification of multiple cancer microarray data sets yields discriminative gene expression signatures that are found and validated on a large number of microarray samples, generated by different laboratories and microarray technologies. Predictive models generated by this approach are better validated than those generated on a single data set, while showing high predictive power and improved generalization performance. PMID- 16271139 TI - Transformation of human bronchial epithelial cells alters responsiveness to inflammatory cytokines. AB - BACKGROUND: Inflammation is commonly associated with lung tumors. Since inflammatory mediators, including members of the interleukin-6 (IL-6) cytokine family, suppress proliferation of normal epithelial cells, we hypothesized that epithelial cells must develop mechanisms to evade this inhibition during the tumorigenesis. This study compared the cytokine responses of normal epithelial cells to that of premalignant cells. METHODS: Short-term primary cultures of epithelial cells were established from bronchial brushings. Paired sets of brushings were obtained from areas of normal bronchial epithelium and from areas of metaplastic or dysplastic epithelium, or areas of frank endobronchial carcinoma. In 43 paired cultures, the signalling through the signal transducer and activator of transcription (STAT) and extracellular regulated kinase (ERK) pathways and growth regulation by IL-6, leukemia inhibitory factor (LIF), oncostatin M (OSM), interferon-gamma (IFNgamma) or epidermal growth factor (EGF) were determined. Inducible expression and function of the leukemia inhibitory factor receptor was assessed by treatment with the histone deacetylase inhibitor depsipeptide. RESULTS: Normal epithelial cells respond strongly to OSM, IFNgamma and EGF, and respond moderately to IL-6, and do not exhibit a detectable response to LIF. In preneoplastic cells, the aberrant signaling that was detected most frequently was an elevated activation of ERK, a reduced or increased IL-6 and EGF response, and an increased LIF response. Some of these changes in preneoplastic cell signaling approach those observed in established lung cancer cell lines. Epigenetic control of LIF receptor expression by histone acetylation can account for the gain of LIF responsiveness. OSM and macrophage-derived cytokines suppressed proliferation of normal epithelial cells, but reduced inhibition or even stimulated proliferation was noted for preneoplastic cells. These alterations likely contribute to the supporting effects that inflammation has on lung tumor progression. CONCLUSION: This study indicates that during the earliest stage of premalignant transformation, a modified response to cytokines and EGF is evident. Some of the altered cytokine responses in primary premalignant cells are comparable to those seen in established lung cancer cell lines. PMID- 16271140 TI - A genome-wide survey of Major Histocompatibility Complex (MHC) genes and their paralogues in zebrafish. AB - BACKGROUND: The genomic organisation of the Major Histocompatibility Complex (MHC) varies greatly between different vertebrates. In mammals, the classical MHC consists of a large number of linked genes (e.g. greater than 200 in humans) with predominantly immune function. In some birds, it consists of only a small number of linked MHC core genes (e.g. smaller than 20 in chickens) forming a minimal essential MHC and, in fish, the MHC consists of a so far unknown number of genes including non-linked MHC core genes. Here we report a survey of MHC genes and their paralogues in the zebrafish genome. RESULTS: Using sequence similarity searches against the zebrafish draft genome assembly (Zv4, September 2004), 149 putative MHC gene loci and their paralogues have been identified. Of these, 41 map to chromosome 19 while the remaining loci are spread across essentially all chromosomes. Despite the fragmentation, a set of MHC core genes involved in peptide transport, loading and presentation are still found in a single linkage group. CONCLUSION: The results extend the linkage information of MHC core genes on zebrafish chromosome 19 and show the distribution of the remaining MHC genes and their paralogues to be genome-wide. Although based on a draft genome assembly, this survey demonstrates an essentially fragmented MHC in zebrafish. PMID- 16271141 TI - Fluid-structure interaction in abdominal aortic aneurysms: effects of asymmetry and wall thickness. AB - BACKGROUND: Abdominal aortic aneurysm (AAA) is a prevalent disease which is of significant concern because of the morbidity associated with the continuing expansion of the abdominal aorta and its ultimate rupture. The transient interaction between blood flow and the wall contributes to wall stress which, if it exceeds the failure strength of the dilated arterial wall, will lead to aneurysm rupture. Utilizing a computational approach, the biomechanical environment of virtual AAAs can be evaluated to study the affects of asymmetry and wall thickness on this stress, two parameters that contribute to increased risk of aneurysm rupture. METHODS: Ten virtual aneurysm models were created with five different asymmetry parameters ranging from beta = 0.2 to 1.0 and either a uniform or variable wall thickness to study the flow and wall dynamics by means of fully coupled fluid-structure interaction (FSI) analyses. The AAA wall was designed to have a (i) uniform 1.5 mm thickness or (ii) variable thickness ranging from 0.5-1.5 mm extruded normally from the boundary surface of the lumen. These models were meshed with linear hexahedral elements, imported into a commercial finite element code and analyzed under transient flow conditions. The method proposed was then compared with traditional computational solid stress techniques on the basis of peak wall stress predictions and cost of computational effort. RESULTS: The results provide quantitative predictions of flow patterns and wall mechanics as well as the effects of aneurysm asymmetry and wall thickness heterogeneity on the estimation of peak wall stress. These parameters affect the magnitude and distribution of Von Mises stresses; varying wall thickness increases the maximum Von Mises stress by 4 times its uniform thickness counterpart. A pre-peak systole retrograde flow was observed in the AAA sac for all models, which is due to the elastic energy stored in the compliant arterial wall and the expansion force of the artery during systole. CONCLUSION: Both wall thickness and geometry asymmetry affect the stress exhibited by a virtual AAA. Our results suggest that an asymmetric AAA with regional variations in wall thickness would be exposed to higher mechanical stresses and an increased risk of rupture than a more fusiform AAA with uniform wall thickness. Therefore, it is important to accurately reproduce vessel geometry and wall thickness in computational predictions of AAA biomechanics. PMID- 16271142 TI - Avoid, attack or do both? Behavioral and physiological adaptations in natural enemies faced with novel hosts. AB - BACKGROUND: Confronted with well-defended, novel hosts, should an enemy invest in avoidance of these hosts (behavioral adaptation), neutralization of the defensive innovation (physiological adaptation) or both? Although simultaneous investment in both adaptations may first appear to be redundant, several empirical studies have suggested a reinforcement of physiological resistance to host defenses with additional avoidance behaviors. To explain this paradox, we develop a mathematical model describing the joint evolution of behavioral and physiological adaptations on the part of natural enemies to their host defenses. Our specific goals are (i) to derive the conditions that may favor the simultaneous investment in avoidance and physiological resistance and (ii) to study the factors that govern the relative investment in each adaptation mode. RESULTS: Our results show that (i) a simultaneous investment may be optimal if the fitness costs of the adaptive traits are accelerating and the probability of encountering defended hosts is low. When (i) holds, we find that (ii) the more that defended hosts are rare and/or spatially aggregated, the more behavioral adaptation is favored. CONCLUSION: Despite their interference, physiological resistance to host defensive innovations and avoidance of these same defenses are two strategies in which it may be optimal for an enemy to invest in simultaneously. The relative allocation to each strategy greatly depends on host spatial structure. We discuss the implications of our findings for the management of invasive plant species and the management of pest resistance to new crop protectants or varieties. PMID- 16271143 TI - Osteoporosis, inflammation and ageing. AB - Osteoporosis is a condition characterized by low bone mass and increased bone fragility, putting patients at risk of fractures, which are major causes of morbidity substantially in older people. Osteoporosis is currently attributed to various endocrine, metabolic and mechanical factors. However, emerging clinical and molecular evidence suggests that inflammation also exerts significant influence on bone turnover, inducing osteoporosis. Numerous proinflammatory cytokines have been implicated in the regulation of osteoblasts and osteoclasts, and a shift towards an activated immune profile has been hypothesized as important risk factor. Chronic inflammation and the immune system remodelling characteristic of ageing, as well as of other pathological conditions commonly associated with osteoporosis, may be determinant pathogenetic factors. The present article will review the current perspectives on the interaction between bone and immune system in the elderly, providing an interpretation of osteoporosis in the light of inflamm-ageing. PMID- 16271144 TI - Reference values for methacholine reactivity (SAPALDIA study). AB - BACKGROUND: The distribution of airway responsiveness in a general population of non-smokers without respiratory symptoms has not been established, limiting its use in clinical and epidemiological practice. We derived reference equations depending on individual characteristics (i.e., sex, age, baseline lung function) for relevant percentiles of the methacholine two-point dose-response slope. METHODS: In a reference sample of 1567 adults of the SAPALDIA cross-sectional survey (1991), defined by excluding subjects with respiratory conditions, responsiveness during methacholine challenge was quantified by calculating the two-point dose-response slope (O'Connor). Weighted L1-regression was used to estimate reference equations for the 95th , 90th , 75th and 50th percentiles of the two-point slope. RESULTS: Reference equations for the 95th , 90th , 75th and 50th percentiles of the two-point slope were estimated using a model of the form a + b* Age + c* FEV1 + d* (FEV1)2 , where FEV1 corresponds to the pre-test (or baseline) level of FEV1. For the central half of the FEV1 distribution, we used a quadratic model to describe the dependence of methacholine slope on baseline FEV1. For the first and last quartiles of FEV1, a linear relation with FEV1 was assumed (i.e., d was set to 0). Sex was not a predictor term in this model. A negative linear association with slope was found for age. We provide an Excel file allowing calculation of the percentile of methacholine slope of a subject after introducing age--pre-test FEV1--and results of methacholine challenge of the subject. CONCLUSION: The present study provides equations for four relevant percentiles of methacholine two-point slope depending on age and baseline FEV1 as basic predictors in an adult reference population of non-obstructive and non atopic persons. These equations may help clinicians and epidemiologists to better characterize individual or population airway responsiveness. PMID- 16271145 TI - A strategy using NMR peptide structures of thromboxane A2 receptor as templates to construct ligand-recognition pocket of prostacyclin receptor. AB - BACKGROUND: Prostacyclin receptor (IP) and thromboxane A2 receptor (TP) belong to rhodopsin-type G protein-coupling receptors and respectively bind to prostacyclin and thromboxane A2 derived from arachidonic acid. Recently, we have determined the extracellular loop (eLP) structures of the human TP receptor by 2-D 1H NMR spectroscopy using constrained peptides mimicking the individual eLP segments. The studies have identified the segment along with several residues in the eLP domains important to ligand recognition, as well as proposed a ligand recognition pocket for the TP receptor. RESULTS: The IP receptor shares a similar primary structure in the eLPs with those of the TP receptor. Forty percent residues in the second eLPs of the receptors are identical, which is the major region involved in forming the ligand recognition pocket in the TP receptor. Based on the high homology score, the eLP domains of the IP receptor were constructed by the homology modeling approach using the NMR structures of the TP eLPs as templates, and then configured to the seven transmembrane (TM) domains model constructed using the crystal structure of the bovine rhodopsin as a template. A NMR structure of iloprost was docked into the modeled IP ligand recognition pocket. After dynamic studies, the segments and residues involved in the IP ligand recognition were proposed. A key residue, Arg173 involved in the ligand recognition for the IP receptor, as predicted from the modeling, was confirmed by site-directed mutagenesis. CONCLUSION: A 3-D model of the human IP receptor was constructed by homology modeling using the crystal structure of bovine rhodopsin TM domains and the NMR structures of the synthetic constrained peptides of the eLP domains of the TP receptor as templates. This strategy can be applied to molecular modeling and the prediction of ligand recognition pockets for other prostanoid receptors. PMID- 16271147 TI - Constitutive degradation of IkappaBalpha in human T lymphocytes is mediated by calpain. AB - BACKGROUND: Activation-induced induction of transcription factor NFkappaB in T lymphocytes is regulated by its inhibitor IkappaBalpha. NFkappaB activation has been demonstrated to occur either by phosphorylation on serine residues 32 and 36 of the inhibitor, IkappaBalpha, followed by ubiquitination and degradation of the inhibitor by the 26S proteasome, or by a proteasome-independent mechanism involving tyrosine phosphorylation, but not degradation. However, the mechanism underlying constitutive regulation of the levels of the inhibitor, IkappaB, in primary human T lymphocytes, remains to be fully delineated. RESULTS: We demonstrate here, the involvement of a proteasome-independent pathway for constitutive regulation of IkappaBalpha levels in primary human T lymphocytes. Pretreatment with a cell permeable calpain inhibitor, E64D, but not with a proteasome specific inhibitor, lactacystin, blocks stimulus-independent IkappaBalpha degradation in primary human T cells. However, E64D pre-treatment fails to impact on IkappaBalpha levels following stimulation with either TNFalpha or pervanadate. Other isoforms of the inhibitor, IkappaBbeta, and IkappaBgamma, appear not to be subject to a similar ligand-independent regulation. Unlike the previously reported decline in ligand-induced degradation of IkappaBalpha in T cells from the elderly, constitutive degradation does not exhibit an age associated decline, demonstrating proteasome-independent regulation of the activity. CONCLUSION: Our studies support a role for an E64D sensitive protease in regulating constitutive levels of IkappaBalpha in T cells, independent of the involvement of the 26S proteasome, and suggests a biological role for constitutive degradation of IkappaBalpha in T cells. PMID- 16271146 TI - Effects of screening and brief intervention training on resident and faculty alcohol intervention behaviours: a pre- post-intervention assessment. AB - BACKGROUND: Many hazardous and harmful drinkers do not receive clinician advice to reduce their drinking. Previous studies suggest under-detection and clinician reluctance to intervene despite awareness of problem drinking (PD). The Healthy Habits Project previously reported chart review data documenting increased screening and intervention with hazardous and harmful drinkers after training clinicians and implementing routine screening. This report describes the impact of the Healthy Habits training program on clinicians' rates of identification of PD, level of certainty in identifying PD and the proportion of patients given advice to reduce alcohol use, based on self-report data using clinician exit questionnaires. METHODS: 28 residents and 10 faculty in a family medicine residency clinic completed four cycles of clinician exit interview questionnaires before and after screening and intervention training. Rates of identifying PD, level of diagnostic certainty, and frequency of advice to reduce drinking were compared across intervention status (pre vs. post). Findings were compared with rates of PD and advice to reduce drinking documented on chart review. RESULTS: 1,052 clinician exit questionnaires were collected. There were no significant differences in rates of PD identified before and after intervention (9.8% vs. 7.4%, p = .308). Faculty demonstrated greater certainty in PD diagnoses than residents (p = .028) and gave more advice to reduce drinking (p = .042) throughout the program. Faculty and residents reported higher levels of diagnostic certainty after training (p = .039 and .030, respectively). After training, residents showed greater increases than faculty in the percentage of patients given advice to reduce drinking (p = .038), and patients felt to be problem drinkers were significantly more likely to receive advice to reduce drinking by all clinicians (50% vs. 75%, p = .047). The number of patients receiving advice to reduce drinking after program implementation exceeded the number of patients felt to be problem drinkers. Recognition rates of PD were four to eight times higher than rates documented on chart review (p = .028). CONCLUSION: This program resulted in greater clinician certainty in diagnosing PD and increases in the number of patients with PD who received advice to reduce drinking. Future programs should include booster training sessions and emphasize documentation of PD and brief intervention. PMID- 16271148 TI - A rigorous method for multigenic families' functional annotation: the peptidyl arginine deiminase (PADs) proteins family example. AB - BACKGROUND: large scale and reliable proteins' functional annotation is a major challenge in modern biology. Phylogenetic analyses have been shown to be important for such tasks. However, up to now, phylogenetic annotation did not take into account expression data (i.e. ESTs, Microarrays, SAGE, ...). Therefore, integrating such data, like ESTs in phylogenetic annotation could be a major advance in post genomic analyses. We developed an approach enabling the combination of expression data and phylogenetic analysis. To illustrate our method, we used an example protein family, the peptidyl arginine deiminases (PADs), probably implied in Rheumatoid Arthritis. RESULTS: the analysis was performed as follows: we built a phylogeny of PAD proteins from the NCBI's NR protein database. We completed the phylogenetic reconstruction of PADs using an enlarged sequence database containing translations of ESTs contigs. We then extracted all corresponding expression data contained in EST database This analysis allowed us 1/To extend the spectrum of homologs-containing species and to improve the reconstruction of genes' evolutionary history. 2/To deduce an accurate gene expression pattern for each member of this protein family. 3/To show a correlation between paralogous sequences' evolution rate and pattern of tissular expression. CONCLUSION: coupling phylogenetic reconstruction and expression data is a promising way of analysis that could be applied to all multigenic families to investigate the relationship between molecular and transcriptional evolution and to improve functional annotation. PMID- 16271149 TI - The effect of motor control exercise versus placebo in patients with chronic low back pain [ACTRN012605000262606]. AB - BACKGROUND: While one in ten Australians suffer from chronic low back pain this condition remains extremely difficult to treat. Many contemporary treatments are of unknown value. One potentially useful therapy is the use of motor control exercise. This therapy has a biologically plausible effect, is readily available in primary care and it is of modest cost. However, to date, the efficacy of motor control exercise has not been established. METHODS: This paper describes the protocol for a clinical trial comparing the effects of motor control exercise versus placebo in the treatment of chronic non-specific low back pain. One hundred and fifty-four participants will be randomly allocated to receive an 8 week program of motor control exercise or placebo (detuned short wave and detuned ultrasound). Measures of outcomes will be obtained at follow-up appointments at 2, 6 and 12 months after randomisation. The primary outcomes are: pain, global perceived effect and patient-generated measure of disability at 2 months and recurrence at 12 months. DISCUSSION: This trial will be the first placebo controlled trial of motor control exercise. The results will inform best practice for treating chronic low back pain and prevent its occurrence. PMID- 16271150 TI - Characterization of SR3 reveals abundance of non-LTR retrotransposons of the RTE clade in the genome of the human blood fluke, Schistosoma mansoni. AB - BACKGROUND: It is becoming apparent that perhaps as much as half of the genome of the human blood fluke Schistosoma mansoni is constituted of mobile genetic element-related sequences. Non-long terminal repeat (LTR) retrotransposons, related to the LINE elements of mammals, comprise much of this repetitive component of the schistosome genome. Of more than 12 recognized clades of non-LTR retrotransposons, only members of the CR1, RTE, and R2 clades have been reported from the schistosome genome. RESULTS: Inspection of the nucleotide sequence of bacterial artificial chromosome number 49_J_14 from chromosome 1 of the genome of Schistosoma mansoni (GenBank AC093105) revealed the likely presence of several RTE-like retrotransposons. Among these, a new non-LTR retrotransposon designated SR3 was identified and is characterized here. Analysis of gene structure and phylogenetic analysis of both the reverse transcriptase and endonuclease domains of the mobile element indicated that SR3 represented a new family of RTE-like non LTR retrotransposons. Remarkably, two full-length copies of SR3-like elements were present in BAC 49-J-14, and one of 3,211 bp in length appeared to be intact, indicating SR3 to be an active non-LTR retrotransposon. Both were flanked by target site duplications of 10-12 bp. Southern hybridization and bioinformatics analyses indicated the presence of numerous copies (probably >1,000) of SR3 interspersed throughout the genome of S. mansoni. Bioinformatics analyses also revealed SR3 to be transcribed in both larval and adult developmental stages of S. mansoni and to be also present in the genomes of the other major schistosome parasites of humans, Schistosoma haematobium and S. japonicum. CONCLUSION: Numerous copies of SR3, a novel non-LTR retrotransposon of the RTE clade are present in the genome of S. mansoni. Non-LTR retrotransposons of the RTE clade including SR3 appear to have been remarkably successful in colonizing, and proliferation within the schistosome genome. PMID- 16271151 TI - Cost and efficiency of public sector sexually transmitted infection clinics in Andhra Pradesh, India. AB - BACKGROUND: Control of sexually transmitted infections (STIs) is an important part of the effort to reduce the risk of HIV/AIDS. STI clinics in the government hospitals in India provide services predominantly to the poor. Data on the cost and efficiency of providing STI services in India are not available to help guide efficient use of public resources for these services. METHODS: Standardised methods were used to obtain detailed cost and output data for the 2003-2004 fiscal year from written records and interviews in 14 government STI clinics in the Indian state of Andhra Pradesh. The economic cost per patient receiving STI treatment was calculated, and the variations of total and unit costs across the STI clinics analysed. Multivariate regression technique was used to estimate incremental unit costs. The optimal number of STIs that could be handled by the clinics was estimated. RESULTS: 18807 STIs were diagnosed and treated at the 14 STI clinics in fiscal year 2003-2004 (range 323-2784, median 1199). The economic cost of treating each STI varied 5-fold from Indian Rupees (INR) 225.5 ( 4.91 US dollars) to INR 1201.5 (26.15 US dollars) between 13 clinics, with one other clinic having a very high cost of INR 2478.5 (53.94 US dollars). The average cost per STI treated for all 14 clinics combined was INR 729.5 (15.88 US dollars). Personnel salaries made up 76.2% of the total cost. The number of STIs treated per doctor full-time equivalent and cost-efficiency for each STI treated had a significant direct non-linear relation (p < 0.001, R2 = 0.81; power function). With a multiple regression model, apart from the fixed costs, the incremental cost for each STI detected and cost of treatment was INR 55.57 (1.21 US dollars) and for each follow-up visit was INR 3.75 (0.08 US dollars). Based on estimates of optimal STI cases that could be handled without compromising quality by each doctor full-time equivalent available, it was projected that at 8 of the 14 clinics substantially more STI cases could be handled, which could increase the total STI cases treated at the 14 clinics combined by 38% at an additional cost of only 3.5% for service provision. CONCLUSION: There is un-utilised capacity in the public sector STI clinics in this Indian state. Efforts to facilitate utilisation of this capacity would be useful, as this would enable more poor patients with STIs to be served at minimal additional cost, and would also reduce the cost per STI treated leading to more efficient use of public resources. PMID- 16271152 TI - Hypercholesterolemia and apolipoprotein B expression: regulation by selenium status. AB - BACKGROUND: Apolipoprotein B (apoB) contains ligand-binding domain for the binding of LDL to LDL-R site, which enables the removal of LDL from circulation. Our recent data showed that selenium (Se) is involved in the lipid metabolism. The present study was aimed to understand the effect of Se deficiency (0.02 ppm) and selenium supplementation (1 ppm) on apoB expression in liver during hypercholesterolemia in male Sprague Dawley rats. Animals were fed with control and high cholesterol diet (2%) for 1 and 2 months. ApoB levels by ELISA and protein expression by western blot was done. Hepatic LDL receptor (LDL-R) activity (in vivo) and mRNA expression by RT-PCR was monitored. RESULTS: In selenium deficiency and on high cholesterol diet (HCD) feeding apoB levels increased and LDL-R expression decreased significantly after 2 months. On 1 ppm selenium supplementation apoB expression significantly decreased and LDL-R expression increased after 2 months. But after one month of treatment there was no significant change observed in apoB and LDL-R expression. CONCLUSION: So the present study demonstrates that Se deficiency leads to up regulation of apoB expression during experimental hypercholesterolemia. Selenium supplementation upto 1 ppm leads to downregulation of apoB expression. Further, this study will highlight the nutritional value of Se supplementation in lipid metabolism. PMID- 16271154 TI - Short-term administration of growth hormone (GH) lowers blood pressure by activating eNOS/nitric oxide (NO)-pathway in male hypophysectomized (Hx) rats. AB - BACKGROUND: The aim of the study was to evaluate the acute and continuous (up to 14 days of treatment) effect of growth hormone (GH) on blood pressure (BP) regulation and to investigate the interplay between GH, nitric oxide (NO) and BP. In un-supplemented and GH supplemented hypophysectomized (Hx) male rats as well as intact rats, continuous resting mean arterial blood pressure (MAP) was measured using telemetry. Baroreceptor activity and the influences of NO on BP control were assessed during telemetric measurement. Furthermore, basal plasma and urine nitrate levels and aortic endothelial nitric oxide synthase (eNOS) expression were analysed. Endothelial function as well as vascular structure in the hindquarter vascular bed was estimated using an in vivo constant-flow preparation. RESULTS: Hypophysectomy was associated with decreased MAP (Hx: 83 +/ 3 vs Intact: 98 +/- 6 mmHg, p < 0.05) and heart rate (HR) (Hx: 291 +/- 4 vs Intact: 351 +/- 7 beat/min, p < 0.05). Endothelial dysfunction and reduced vasculature mass in the hindquarter vascular bed was found in Hx rats. GH substitution caused a further transient decrease in MAP and a transient increase in HR (14% and 16% respectively, p < 0.05). The reduction in MAP appeared to be NO dependent. Aortic eNOS expression was unchanged. GH substitution resulted in an impaired baroreceptor function. Two weeks of GH treatment did not normalise the BP, vascular structure and the endothelial function in the resistance vessels. CONCLUSION: GH substitution seems to have a short lasting effect on lowering blood pressure via activation of the NO-system. An interaction between GH, NO-system and BP regulation can be demonstrated. PMID- 16271153 TI - Malaria chemoprophylaxis and the serologic response to measles and diphtheria tetanus-whole-cell pertussis vaccines. AB - BACKGROUND: Acute malaria has been associated with a decreased antibody response to tetanus and diphtheria toxoids, meningococcal, salmonella, and Hib vaccines. Interest in giving malaria drug therapy and prevention at the time of childhood immunizations has increased greatly following recent trials of intermittent preventive therapy during infancy (IPTi), stimulating this re-analysis of unpublished data. The effect of malaria chemoprophylaxis on vaccine response was studied following administration of measles vaccines and diphtheria-tetanus-whole cell pertussis (DTP) vaccines. METHODS: In 1975, six villages divided into two groups of children < or = 74 months of age from Burkina Faso, were assigned to receive amodiaquine hydrochloride chemoprophylaxis (CH+) every two weeks for seven months or no chemoprophylaxis (CH-). After five months, children in each group received either one dose of measles or two doses of DTP vaccines. RESULTS: For recipients of the measles vaccine, the seroconversion rates in CH+ and CH- children, respectively, were 93% and 96% (P > 0.05). The seroresponse rates in CH+ and CH- children respectively, were 73% and 86% for diphtheria (P > 0.05) and 77% and 91% for tetanus toxoid (P > 0.05). In a subset analysis, in which only children who strictly adhered to chemoprophylaxis criteria were included, there were, likewise, no significant differences in seroconversion or seroresponse for measles, diphtheria, or tetanus vaccines (P > 0.05). While analysis for pertussis showed a 43% (CH+) and 67% (CH-) response (P < 0.05), analyses using logistic regression to control for sex, age, chemoprophylaxis, weight-for-height Z-score, and pre-vaccination geometric mean titer (GMT), demonstrated that chemoprophylaxis was not associated with a significantly different conversion rate following DTP and measles vaccines. Seven months of chemoprophylaxis decreased significantly the malaria IFA and ELISA GMTs in the CH+ group. CONCLUSION: Malaria chemoprophylaxis prior to vaccination in malaria endemic settings did not improve or impair immunogenicity of DTP and measles vaccines. This is the first human study to look at the association between malaria chemoprophylaxis and the serologic response to whole-cell pertussis vaccine. PMID- 16271155 TI - Downregulation of peroxisome proliferator-activated receptors (PPARs) in nasal polyposis. AB - BACKGROUND: Peroxisome proliferator-activated receptor (PPAR) alpha, betadelta and gamma are nuclear receptors activated by fatty acid metabolites. An anti inflammatory role for these receptors in airway inflammation has been suggested. METHODS: Nasal biopsies were obtained from 10 healthy volunteers and 10 patients with symptomatic allergic rhinitis. Nasal polyps were obtained from 22 patients, before and after 4 weeks of local steroid treatment (fluticasone). Real-time RT PCR was used for mRNA quantification and immunohistochemistry for protein localization and quantification. RESULTS: mRNA expression of PPARalpha, PPARbetadelta, PPARgamma was found in all specimens. No differences in the expression of PPARs were obtained in nasal biopsies from patients with allergic rhinitis and healthy volunteers. Nasal polyps exhibited lower levels of PPARalpha and PPARgamma than normal nasal mucosa and these levels were, for PPARgamma, further reduced following steroid treatment. PPARgamma immunoreactivity was detected in the epithelium, but also found in smooth muscle of blood vessels, glandular acini and inflammatory cells. Quantitative evaluation of the epithelial immunostaining revealed no differences between nasal biopsies from patients with allergic rhinitis and healthy volunteers. In polyps, the PPARgamma immunoreactivity was lower than in nasal mucosa and further decreased after steroid treatment. CONCLUSION: The down-regulation of PPARgamma, in nasal polyposis but not in turbinates during symptomatic seasonal rhinitis, suggests that PPARgamma might be of importance in long standing inflammations. PMID- 16271157 TI - Adjunctive corticosteroids for Pneumocystis jiroveci pneumonia in patients with HIV infection: a meta-analysis of randomised controlled trials. AB - BACKGROUND: The objective of this study was to review the effects of adjunctive corticosteroids on overall mortality and the need for mechanical ventilation in HIV-infected patients with Pneumocystis jiroveci pneumonia (PCP) and substantial hypoxemia (arterial oxygen partial pressure <70 mmHg or alveolar-arterial gradient >35 mmHg on room air). METHODS: We conducted a systematic search of the literature for randomised trials published up to December 2004. Selected trials compared adjunctive corticosteroids with placebo or usual care in HIV-infected patients with PCP and reported mortality data. Two teams of reviewers independently evaluated the methodology and extracted data from each primary study. RESULTS: Six studies were included in the meta-analysis. Risk ratios for overall mortality for adjunctive corticosteroids were 0.54 (95% confidence interval [CI], 0.38-0.79) at 1 month and 0.67 (95% CI, 0.49-0.93) at 3-4 months of follow-up. Numbers needed to treat, to prevent 1 death, are 9 patients in a setting without highly active antiretroviral therapy (HAART) available and 22 patients with HAART available. Only the 3 largest trials provided data on the need for mechanical ventilation with a risk ratio of 0.37 (95% CI, 0.20-0.70) in favour of adjunctive corticosteroids. CONCLUSION: The number and size of trials investigating adjunctive corticosteroids for HIV-infected patients with PCP is small, but our results suggest a beneficial effect for patients with substantial hypoxemia. PMID- 16271158 TI - Reviving a neglected celestial underwater polarization compass for aquatic animals. AB - Substantial in situ measurements on clear days in a variety of marine environments at depths in the water down to 200 m have demonstrated the ubiquitous daytime presence of sun-related e-vector (=plane of polarization) patterns. In most lines of sight the e-vectors tilt from horizontal towards the sun at angles equal to the apparent underwater refracted zenith angle of the sun. A maximum tilt-angle of approximately 48.5 degrees , is reached in horizontal lines of sight at 90 degrees to the sun's bearing (the plane of incidence). This tilt limit is set by Snell's window, when the sun is on the horizon. The biological literature since the 1980s has been pervaded with assumptions that daytime aquatic e-vectors are mainly horizontal. This review attempts to set the record straight concerning the potential use of underwater e-vectors as a visual compass and to reopen the field to productive research on aquatic animals' orientation and navigation. PMID- 16271156 TI - Use of endogenous signal sequences for transient production and efficient secretion by moss (Physcomitrella patens) cells. AB - BACKGROUND: Efficient targeting to appropriate cell organelles is one of the bottlenecks for the production of recombinant proteins in plant systems. A common practice is to use the native secretory signal peptide of the heterologous protein to be produced. Though general features of secretion signals are conserved between plants and animals, the broad sequence variability among signal peptides suggests differing efficiency of signal peptide recognition. RESULTS: Aiming to improve secretion in moss bioreactors, we quantitatively compared the efficiency of two human signal peptides and six signals from recently isolated moss (Physcomitrella patens) proteins. We therefore used fusions of the different signals to heterologous reporter sequences for transient transfection of moss cells and measured the extra- and intracellular accumulation of the recombinant proteins rhVEGF and GST, respectively. Our data demonstrates an up to fivefold higher secretion efficiency with endogenous moss signals compared to the two utilised human signal peptides. CONCLUSION: From the distribution of extra- and intracellular recombinant proteins, we suggest translational inhibition during the signal recognition particle-cycle (SRP-cycle) as the most probable of several possible explanations for the decreased extracellular accumulation with the human signals. In this work, we report on the supremacy of moss secretion signals over the utilised heterologous ones within the moss-bioreactor system. Though the molecular details of this effect remain to be elucidated, our results will contribute to the improvement of molecular farming systems. PMID- 16271159 TI - Progress of leukoaraiosis is inhibited by correction of platelet hyper aggregability. AB - BACKGROUND: Platelet hyper-aggregability is an important risk factor for leukoaraiosis. In this study we investigated whether aggravation of leukoaraiosis can be controlled by means of long-term correction of platelet hyper aggregability. METHODS: Twenty-one patients with leukoaraiosis and uncorrected platelet hyper-aggregability were compared with 21 controls matched for age, grade of leukoaraiosis and observation period whose platelet hyper-aggregability was corrected. Platelet aggregability was estimated by an optical analytical method with a nine-stage display using two different concentrations each of adenosine diphosphate (ADP) and collagen (the double ADP method). RESULTS: The mean observation period between two magnetic resonance imaging (MRI) scans for both groups was 4.1 years. In the non-corrected group, moderate to severe aggravation of leukoaraiosis was observed in a large number of patients. In the corrected group, only a small number of patients showed generally mild aggravation of leukoaraiosis. The number of patients showing aggravation of periventricular hyperintensity (PVH) was 7 in 21 in the non-corrected group versus 1 in 21 (p = 0.022) in the corrected group, and for aggravation of deep white-matter hyperintensity, these values were 9 in 21 versus 4 in 21, respectively. Thus, the difference was more significant if the degree of aggravation was taken into account. CONCLUSION: The progress of leukoaraiosis is greatly inhibited by long-term correction of platelet hyper-aggregability. PMID- 16271160 TI - Hormone and lipolytic responses to whole body vibration in young men. AB - This study examined the effects of whole-body vibration (WBV) on the hormone and lipolytic responses. Eight male subjects performed WBV and control (CON) trials on separate days. The WBV session consisted of 10 sets of vibration for a duration of 60 s with rest periods of 60 s between each set (frequency 26 Hz). The subjects maintained a static squat position with knees bent on the platform. In the CON trial, the WBV stimulation was not imposed. Blood samples were collected before both trials and during the recovery period. In the WBV trial, the concentrations of plasma epinephrine (Epi) and norepinephrine (NE) increased immediately after the session (P < 0.05). Serum free fatty acids (FFA) concentration increased significantly at the 150, 180, and 210 min points of the recovery period in the WBV trial (P < 0.01) with the interaction between trial and time (P < 0.01). Serum glycerol showed no significant change in either trial. These results suggest that the WBV session causes secretions of Epi and NE, and it subsequently increases FFA concentration during the recovery period. However, because the FFA response was inconsistent with that of glycerol, we were unable to clarify the effect of WBV exposure on lipolysis. PMID- 16271161 TI - Preventive and interceptive orthodontic treatment needs of an inner-city group of 6- and 9-year-old Canadian children. AB - OBJECTIVE: Early recognition of developing malocclusions and the potential for uncomplicated orthodontic treatment procedures can minimize or eliminate future costly treatment. This study was designed to assess the potential for this approach in children living in a limited-income environment. A modified index for preventive and interceptive orthodontic needs (IPION) was used to determine the need for such treatment in schoolchildren aged 6 and 9 years. METHODS: Two calibrated examiners examined each child independently and assessed several components of his or her occlusion, including molar relationship, crossbite, open bite, overbite and overjet. Dental variables such as presence of caries and early loss of teeth were also noted. Informed consent was obtained and all children present at school on the day of the field study were included. A total of 395 children were divided into 2 groups, aged 6 and 9 years. RESULTS: A high prevalence of caries in the deciduous dentition (30.4% for 6 year olds; 20.6% for 9 year olds) and early loss of primary teeth (11.9% for 6 year olds; 29.4% for 9 year olds) was observed. A large percentage of children had crossbite in the anterior or posterior segments, or both. Open bites were also a common finding. Future orthodontic problems were identified in 28% of this population by using the modified IPION. No statistically significant differences (p > 0.05) were found between sexes or age groups using the chi2 test. CONCLUSIONS: Most of the developing malocclusions identified in this study would be amenable to interceptive orthodontics, consisting of space maintenance, crossbite correction and arch expansion. PMID- 16271162 TI - Introducing digital radiography in the dental office: an overview. AB - Digital radiography is gaining in popularity and many dentists are considering changing from a film-based system to this new technology. Dentists must clearly define their objectives for adopting digital radiography and be aware of the problems that may be encountered with this equipment, so that they can make an informed purchasing decision. This article provides an overview of digital radiographic equipment, experiences users have had with this technology and factors to consider when deciding to purchase a DR system. PMID- 16271163 TI - The use of lasers for periodontal debridement: marketing tool or proven therapy? AB - The use of lasers in dentistry has recently received much attention, in both clinical practice and research; their unique properties produce favourable clinical results in some cases and encourage patient acceptance. Various types of lasers have been investigated as an adjunct to periodontal therapy; these include carbon dioxide (CO2), diode, neodymium:yttrium-aluminium-garnet (Nd:YAG) and erbium:yttrium-aluminium-garnet (Er:YAG) lasers.However, adverse results have been associated with each type, including thermal damage to root surfaces, increases in pulpal temperature and the production of toxic by products. The Er:YAG laser has produced the most promising results, as it can ablate effectively with minimal adverse effects. More research is needed to determine the ideal settings and methods for using the laser safely and effectively in clinical practice. PMID- 16271164 TI - Treatment of self-inflicted oral trauma in a comatose patient: a case report. AB - Trauma to oral soft tissues in comatose patients may be more widespread than reported, as no extensive study of this problem has been conducted. Various appliances for the prevention of self-inflicted injuries to oral tissues particularly in children and the physically and mentally challenged have been documented, but there is little information on their use in adult comatose patients. Because comatose patients lack cerebral control of the masticatory cycle, they can easily injure themselves. Although it is not uncommon for patients with a decreased level of consciousness and in need of intensive care to be restrained to prevent injury due to involuntary movement of the limbs, head and neck restraint is often difficult and may be dangerous to the patient. This case report presents a simple solution to the problem of self-inflicted trauma to oral tissues. PMID- 16271165 TI - Pemphigus vulgaris: a case-based update. AB - Pemphigus vulgaris (PV) is an autoimmune disease accounting for 80% of all cases of pemphigus. Before the advent of corticosteroid therapy, pemphigus was fatal, with a mortality rate of up to 75% in the first year. It is still a serious disorder, but the 5% to 10% mortality rate is now primarily due to the side effects of therapy. In 75% to 80% of cases, PV lesions first appear in the oral cavity. Dentists are therefore in a unique position to recognize the oral manifestations of the disease, allowing early diagnosis and initiation of treatment. The diagnosis is based on pathological examination and immunofluorescence testing. Systemic corticosteriods and steroid-sparing agents are the mainstays of treatment; topical corticosteroids may also be used to accelerate healing of persistent oral lesions. This article describes a 71-year old woman with multiple chronic ulcers in the oral cavity, in whom PV was diagnosed 4 months after the symptoms first appeared. The article also reviews the current literature on diagnosis and treatment of the condition. PMID- 16271166 TI - How predictable are periodontal regenerative procedures? AB - Periodontal regeneration has become one of the primary objectives of periodontal therapy. The resulting scientific endeavours have elucidated modes of periodontal wound healing, the growth of periodontal cells and their association with the surrounding matrix, and growth-promoting factors. The periodontal regeneration industry is producing better and more expensive devices, but the criteria for evaluating their success have not progressed to the same extent. Although clinical measurements of attachment level and probing depths, along with radiography, are good methods of evaluating tooth survival and prognosis, they do not indicate true biological regeneration. In addition, the regeneration industry may encourage the overuse of allografts and alloplasts which may serve as an impediment to simple wound healing. This review is a critical assessment of the clinical use of various regenerative tools, specifically bone replacements and membranes. The future of the regeneration industry may depend on the merging of various technologies and biological concepts, including the possible use of biological barriers, various bone and periodontal growth inducers, and artificial matrices that will attract or carry the cells necessary for regeneration. PMID- 16271167 TI - [Achieving pathogenesis understanding of ocular diseases by deciphering the underlying molecular pathways]. AB - The field of ophthalmology research has experienced a revolution since the 1970's, when molecular biology techniques were gradually and widely adopted. Many of the developments generated impact that went far beyond the field of ophthalmology. A classic case was the identification and characterization of the Retinoblastoma susceptibility gene (Rb), whose impact went far beyond the rare and obscure disease, as it provides key evidence for the concept of tumor suppressor gene and the "two hit" theory of tumor formation. The identification of scores of genes involved in retinitis pigmentosum (pigmentosa), on the other hand, show cases the complexity of multi-factorial diseases. Ophthalmology researchers in China have been quick in integrating these novel tools into their research. However, the field still lags behind in the effective use of these technologies to carry out in-depth inquiries into key disease mechanisms. The advent of "omics" technologies heralded a new era in biomedical research that allows for the global and rapid survey of genetic and biochemical profiles. Effective integration of these novel technologies into ophthalmology research will have far-reaching impact for the whole field. PMID- 16271168 TI - [Evaluation of classification and treatment for orbital malignant lymphoma]. AB - Orbital malignant lymphoma is a common orbital tumor. Because various types of lymphomas have different degrees of malignancy and clinical manifestations, and each type has a distinct treatment and prognosis, the classification of orbital malignant lymphoma is very important. Up to now the opinions on the classification of orbital lymphoma have been conflicting. We collected related materials from abroad and domestic literature, and summarized our experience of clinical and pathologic work on orbital malignant lymphoma. The work was directed by the histopathologists. Here we propose our classification and treatment protocol of orbital malignant lymphoma. This classification is a tentative one, which needs the comments of our ophthalmic colleagues. PMID- 16271169 TI - [Clinical and pathological features of 112 cases with ocular adnexal lymphoproliferative lesions]. AB - OBJECTIVE: To investigate the histopathological classification and clinical features of ocular adnexal lymphoproliferative lesions. METHODS: The clinical, histomorphological and immuno-histochemical features of 112 cases of ocular adnexal lymphoproliferative lesions (116 paraffin specimens) were studied retrospectively. The lesions were classified according to the World Health Organization classification of tumors of haematopoietic and lymphoid tissues (2001). RESULTS: This group of patients' ages averaged at 49. The mean duration between the onset of the symptoms and the time of presentation was 22 months. Sixteen patients (14.3%) had bilateral lesions. Proptosis or local orbital mass was presented in 69 cases (61.6%). Reactive lymphoid hyperplasia was diagnosed in 11 cases (9.8%) and atypical lymphoid hyperplasia in 10 cases (8.9%). The rest 91 cases (81.3%) were diagnosed as lymphoma, among which 74 cases (81.3%) were extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue (MALT-EMZL). CONCLUSIONS: MALT-EMZL is the most frequent type of ocular lymphoproliferative lesions in ocular adnexa. The ocular adnexal lymphoproliferative lesions show an indolent course clinically, tending to affect the old. Bilateral case is not uncommon. PMID- 16271170 TI - [Clinical and pathological analysis of 151 cases of tumors in lacrimal fossa]. AB - OBJECTIVE: To investigate the clinical features and pathological classification of tumors in lacrimal fossa. METHODS: A retrospective study of 151 cases of tumors in lacrimal fossa in recent 8 years from EENT hospital was performed to investigated the clinical features, histopathological classification and prognoses. RESULTS: In 69 cases of benign lacrimal epithelial tumors, pleomorphic adenoma was the most common tumor (63 cases, 41.7%), followed by myoepithelioma (3 cases, 1.2%), lacrimal duct cyst (2 cases, 1.3%). In 34 cases of malignant lacrimal epithelial tumors, adenoid cystic carcinoma was the most common tumor (15 cases, 9.9%), followed by pleomorphic adenocarcinoma (14 cases, 9.3%), adenocarcinoma (4 cases, 2.6%). There were 48 cases of lacrimal non-epithelial lesions. Mucosa-associated lymphoid tissue (MALT) lymphoma was the most common tumor (17 cases, 11.3%), followed by pseudotumor (12 cases, 7.9%), benign lymphoepithelial lesion (7 cases, 4.0%). CONCLUSIONS: There are a variety of tumors in lacrimal fossa. The treatment of pleomorphic adenoma and the diagnosis of MALT lymphoma are the most difficult tasks in present clinical work. PMID- 16271171 TI - [Transpupillary thermotherapy for circumscribed choroidal hemangioma]. AB - OBJECTIVE: To evaluate the efficacy of transpupillary thermotherapy (TTT) on circumscribed choroidal hemangioma. METHODS: Ten eyes of 10 cases with circumscribed choroidal hemangioma (age 16-48, average 34.4) were included in the present study. Six cases had previous green laser (4 argon and 2 krypton) photocoagulation, TTT as a primary treatment was used for other 4 cases without previous laser application. Fluorescein angiography, visual acuity, anterior segment and fundus examination, perimetry test, FFA/ICGA and ultrasonography were performed in all patients before the treatment. One to five infra red diode laser spots (810 nm with power of 800-1200 mW, diameter of 3 mm) were delivered on the surface of tumor with 60 seconds exposure, additional 10-20 seconds exposure was used in some cases. Second treatment was given 1-3 month later if there was active leakage demonstrated by fluorescein angiography. The followed-up period was 6-36 months (averaged 14.5 month). RESULTS: After transpupillary thermotherapy, 9 eyes (90%) showed partial regression of the tumor. Average visual acuity was 0.25 and 0.6 before and after TTT, respectively. Six patients showed improvement of vision acuity, but vision acuity in 4 patients with cystic macular degeneration remained unchanged. No recurrences were observed within follow-up averaged 14.5 months. Transpupillary thermotherapy did not associate with any significant complications. CONCLUSION: Transpupillary thermotherapy is effective to treat circumscribed hemangioma of choroid either as a primary treatment or supplementary therapy to previous laser photocoagulation. PMID- 16271172 TI - [In vitro study on inhibition of human choroidal melanoma OCM-1 cell line by arsenic trioxide]. AB - OBJECTIVE: To investigate the effects and mechanisms of arsenic trioxide (ATO) on human choroidal melanoma cell line OCM-1. METHODS: OCM-1 cells were cultured with 0.75 to 24.00 micromol/L arsenic trioxide for various durations, then cell viability was measured by MTT assay. The cell necrosis and apoptosis rates were observed by flow cytometry. The morphological changes of the cells were examined by electron microscopy. Glutathione peroxidase (GSH-Px) activities were tested. Mitochondrial membrane potential (MMP) was detected by confocal microscopy. RESULTS: Growth of OCM-1 cells was inhibited by ATO at concentrations of (1.5 to 24.0) micromol/L. However, there was no effect of 0.75 micromol/L ATO on the growth of OCM-1 cells. The inhibition showed both dose and time dependent effects (P < 0.05). The IC(50) was 16.8 micromol/L at 24 h. Flow cytometry analysis showed a positive correlation between the rate of cell necrosis and apoptosis and the concentration of ATO. The cell necrosis rates were higher than the cell apoptosis rates at various concentrations of ATO. OCM-1 cells cultured with ATO showed the classic morphologic characteristics of necrosis and apoptosis. GSH-Px activities and MMP decreased in a dose dependent manner. CONCLUSION: ATO inhibits the growth of OCM-1 cells. The mechanism of this effect is that ATO inhibits the GSH-Px activities, decreases the MMP and impairs mitochondrial energy synthesis, which induces necrosis and apoptosis of human choroidal melanoma COM-1 cells eventually. PMID- 16271173 TI - [The effect of Chitosan on intraocular structure when filling in the vitreous cavity]. AB - OBJECTIVE: To study the effect of Chitosan on intraocular structure and pressure when filling in the vitreous body. METHODS: Chitosan were injected into vitreous cavity in twelve healthy vitrectomized pigmented rabbits in the right eyes (experiment group) and Hyalumnan in the left eyes (control group). Slitlamp biomicroscopy and indirect ophthalmoscopy were performed and intraocular pressure were measured pre- and postoperatively in all eyes. Histopathological examination was done after the rabbits were sacrificed at the end of the study. RESULTS: All the results indicated that there were no significant differences between experiment group and control group. CONCLUSION: There is no obvious effect on intraocular structure and intraocular pressure when the vitreous cavity is filled with Chitosan. PMID- 16271174 TI - [Comparison on morphology and ultrastructure of two kinds of myopic models]. AB - OBJECTIVE: To study the possibility of inducing refractive errors by opening the mouse (C57BL/6J) eye artificially after birth and explore the difference in pathogenic mechanism in form-deprivation myopia by analyzing the histological changes of the animal eyes. METHODS: 4-day-old mice (n = 13) was undergone 10-day monocular artificially opened period till the opposite eye was natural open. Another group 12-day-old mice (n = 13) were raised with monocular suture of eyelid for 7 days. Age-matched untreated mice were used as control. Retinoscopic refraction was performed subsequently. Animals were sacrificed and the eyes enucleated at the end of the experiment. The axial length and weight of the eyes were measured and the sections of the eyes were also observed by light and electron microscope. RESULTS: All eyes treated with two methods showed significant development of myopia compared with the contralateral eye. The axial length was elongated in form deprived eyes, but it was shortened in artificially opened eyes; no change in the weight of eyeball was found in form deprived eyes, but it was reduced in artificially opened eyes when compared with corresponding fellow eyes. Under light microscope, there is no significant histological difference observed in the eyes treated with two methods. Under electron microscope, rod outer segment in form deprived eyes was inordinate and the phagosomes containing fractions of the membrane disc of outer segment were remarkably decreased. No significant changes of photoreceptor were demonstrated in artificially opened eyes. CONCLUSION: The means of artificially opening eyelid can induce myopia in the mouse and the method may be useful to establish myopia animal model. PMID- 16271175 TI - [Tumor targeting gene transfer mediated by electroporation for growth suppression of retinoblastoma in vivo]. AB - OBJECTIVE: To explore the effects and mechanisms of electroporation on the therapy of transplanted retinoblastoma tumor (RB) by transferring the expressive plasmids of sFlk-1 and ExTek. METHODS: The effects and mechanisms of sFlk-1 and ExTek on RB is validated both in vitro and in vivo. In vitro, pCI-sFlk-1 was transfected into HEK293 (human embryonic kidney 293) cells with DMRIE-C Reagent, and its supernatant was collected 24 h later to mix with RPMI-1640 medium as 1:1, 2:1 and 3:1. RB cells were cultured in the mixed medium and the cell growth curve were analyzed 6 days later. The suspension of 1 x 10(7) RB cells were inoculated in 40 nude mice subcutaneously. The nude mice with RB were divided into 2 groups. The experimental group was treated with transfer of pCI-sFlk-1 (15 microg) and pCI-ExTek (15 microg) by electroporation once a week, while the control group was treated with physiological saline solution. After 3 weeks' therapy, the animals were observed for one more week. During this period the tumors were measured to draw the growth curve of the tumors and to calculate the anti-tumor rate. The vessel endothelium were tagged with smooth muscle actin (SMA) by immunohistochemical methods to inspect the micro vessel density (MVD) of the tumors. Moreover, the expression of vascular endothelial growth factor (VEGF) and its receptor Flk-1 of the tumors was confirmed by immunohistochemical staining. RESULTS: sFlk-1 exhibited no inhibiting effects on the growth of RB cells in vitro, while sFlk-1 combined with ExTek restrained the growth of the transplanted tumors obviously in vivo, and the anti-growth rate is 80.04%. The MVD of the experimental group is 26.69 +/- 2.95, which is evidently lower than that of the control group (44.51 +/- 6.11). The difference between these 2 groups was statistically significant (P < 0.05). As to the expression of VEGF, there was no significant difference between these 2 groups, while the expression of Flk-1 of the experimental group was much stronger than that of the control group. CONCLUSION: Transfer the expressive plasmids of sFlk-1 and ExTek by electroporation in vivo restrained the growth of the transplanted RB tumor, and the effect is due to the inhibition of angiogenesis process. PMID- 16271176 TI - [Variety of differential display on gene and corresponding gene's clone following corneal alkali burns in rats]. AB - OBJECTIVE: To study the condition of differential gene caused after corneal alkali burns in rats and clarify the molecular biological foundation of corneal denatured protein. METHODS: The animals were sacrificed on the 3rd day and the 2nd week after alkali burns. Total RNA was isolated from the excised corneas and then reverse-transcribed into cDNA. The differential gene was detected by mRNA differential display reverse transcription polymerase chain reaction (DDRT-PCR) with two kinds of anchoring primer and 12 kinds of random primers after corneal alkali burns in the rats. The differential gene fragments were cloned, and their homogeneity was compared with each other in the Gene Bank. RESULTS: Compared with the normal cornea, the cornea of alkali burns on the 2nd week produced one differential gene fragment of 630 bp in the same reaction condition, and this differential gene was homologous to the rattus norvegicus mitochondrial cytochrome oxidase subunits I, II, III gene. CONCLUSIONS: It is found that there is differential gene in the cornea of alkali burns on the 2nd week in rats, and this differential gene is homologous to the rattus norvegicus mitochondrial cytochrome oxidase subunits I, II, III gene. It can be concluded that the occurrence of this differential gene is possible to be related with the action of the superoxide free radicals caused by alkali burns. PMID- 16271177 TI - [Measurement of macular edema in retinal vein occlusion using optical coherence tomography and its visual prognosis]. AB - OBJECTIVE: To examine the type of macular edema in patients with retinal vein occlusion by using optical coherence tomography (OCT). To compare the sensitivity and specificity between OCT and fundus fluorescein angiography (FFA). To investigate the visual prognosis and risk factors in patients with cystoid macular edema (CME). METHODS: Ninety-one eyes of 90 patients with various types of retinal vein occlusion were examined by OCT and FFA. There were 54 cases male and 36 cases female. The age of these patients ranged from 20 to 74 years old and averaged (57.8 +/- 13.8) years old. Right eye was affected in 45 cases, and left eye in 46 cases (both eye in 1 case). Central retinal vein, hemicentral retinal vein and branch retinal vein were affected in 54, 9 and 28 eyes, respectively. The average follow-up period was 6.10 months. The sensitivity and specificity of OCT and FFA were measured. The visual prognosis and risk factors were analyzed. Macular central retinal thickness of 54 opposite normal eye was measured for comparison. RESULTS: (1) The classification of macular edema by OCT was cystoid macular edema in 71 eyes (78.0%); subretinal fluid in 14 eyes and laminar macular hole in 1 eye. The minimal and maximal height of intraretinal cystoid space was 94 microm and 1317 microm, respectively and averaged (668.18 +/- 245.58) microm. The minimal and maximal height of macular central retinal thickness of 54 opposite normal eye was 110 microm and 236 microm, respectively and averaged (154.09 +/- 21.85) microm. The maximal height of subretinal fluid space was 1377 microm or even beyond the detective limit of OCT. (2) The sensitivity of OCT for detection of CME was 98.6% and the specificity was 100%. The sensitivity of FFA was 86.1% and the specificity was 100.0%. (3) The visual prognosis of 61 eyes with CME follow-up over 3 months: The difference between the initial and final VA in branch retinal vein occlusion was statistically significant (P < 0.01), while no difference between initial and final VA in central and hemicentral retinal vein occlusion (P > 0.05). The factors affected visual prognosis of CME included the duration, the presence of hemorrhage in cystoid space and the severity of occlusion, etc. In CME under 3 months, the visual prognosis after therapy is better than that of before the therapy (P < 0.01); while in eyes with duration more than 3 months, no difference of vision could be detected before and after the therapy (P > 0.05). CONCLUSIONS: OCT is a safe, high-resolution, non invasive, reliable and reproducible new examine method for detecting CME. The visual prognosis of CME is poor. It is possible that the visual prognosis can be improved by early detecting CME using OCT. PMID- 16271178 TI - [Ocular features of polyneuropathy-organomegaly-endocrinopathy-monoclonal gammopathy-skin changes syndrome]. AB - OBJECTIVE: To analyze the ocular features of POEMS syndrome, and explore the pathogenesis. METHOD: Fifteen cases with POEMS syndrome from 1984-2004 were reviewed and summarized. The ocular features and the intracranial pressures were analyzed. RESULTS: Three of 15 patients showed blurred vision. Seven of 15 patients had mild papilloedema, five had fully papilloedema, and the optic discs in other three cases were normal. Intracranial pressures were measured in 11 of 15 patients. Seven of 8 patients with papilloedema had elevated intracranial pressures. CONCLUSIONS: Papilloedema is found in about 80.0% patients with POEMS syndrome. Most of the patients with papilloedema have elevated intracranial pressures. PMID- 16271179 TI - [In vivo confocal microscopy observation and evaluation of sensation in diabetic corneas]. AB - OBJECTIVE: To test the changes of corneal sensation in diabetic patients and to observe the diabetic corneas using confocal microscopy. METHODS: One hundred and thirty-two diabetic patients were enrolled in this study. The diabetic history and medication were recorded. The FBG and HbA(1c) were checked for every patient. The patient's eye was systemically checked, including the IOP, visual acuity, cornea sensation, lenses, and the fundus. Two hundred patients of other diseases were enrolled as controls. The cornea of 85 patients and 56 controls were checked by the confocal microscopy. The right eyes of both patients and controls were used for analysis. RESULTS: The cornea sensation of diabetic patients was significantly depressed as compared to that of the controls (P < 0.01). The depression of cornea sensation was related to the course of the disease, the presence of diabetic retinopathy and the degree of the diabetic retinopathy (P < 0.05). The confocal microscopy revealed that there was a higher corneal epithelium edema rate (11.76%) in diabetic patients than that of the controls (1.79%) (P < 0.05). The nerve fiber density of the diabetic patients was decreased and there were some morphological changes in the nerve fiber of diabetic patients, such as sparse, slim, less branches, less long fibers, edema and zigzag of the fibers. CONCLUSION: The depression of the corneal sensation in diabetic patients is probably caused by the reduction and the degeneration of the corneal nerve fibers. PMID- 16271180 TI - [Studies on vision mediated behavioral changes of rat with photic injury]. AB - OBJECTIVE: To evaluate the relationship of visual function and photoreceptor cell loss, and to establish a visual function assessment system based on the observation of optokinetic behavior in rat with photic injury. METHODS: SD rats were exposed to (950 +/- 50) lux green light for 3 hours, 12 hours and 24 hours respectively after 24-hour dark adaptation. Then 24-hour dark recovery was followed by 48-hour observation before the eyes were enucleated. Frozen sections were obtained. Slides were stained by HE and rhodopsin immunofluorescent stainings. Vision mediated disturbance of consciousness and pre-pulse response were used to assess the visual function. Optokinetic behavior test was conducted 2 days before and after photic injury. Average velocity of motion were analyzed. RESULTS: Vision mediated disturbance of consciousness response are gradually weakened; while pre-pulse response is reinforced as the photic injury time increases. The relationship of optokinetic behavior and rat visual function is statistically significant. Optokinetic behavior test shows the average velocity of motion gradually decreases as the injury is aggravated. CONCLUSIONS: Longer light exposure time leads to increased photoreceptor injury, which in turn, causes decrease of visual function in SD rat. Optokinetic behavior test, represented by average velocity of motion, is correlated with rat visual function. Thus, optokinetic behavior test can be used to assess the visual function of SD rat quantitatively. PMID- 16271181 TI - [Experimental study on the effects of rapamycin in prevention of rat corneal allograft rejection]. AB - OBJECTIVE: To investigate the effects of rapamycin on prevention of corneal allograft rejection in murine corneal transplantation. METHODS: The outbred strain SD and Wistar rats were used as donors and recipients, respectively. Sixty eight Wistar rats were divided into four groups: Group A, autograft control; Group B, allograft control (the control groups were given placebo only); Group C and D, allograft groups, were treated with orally RAPA (3 mg.kg(-1).d(-1)) and CsA (10 mg.kg(-1).d(-1)), respectively. The drugs were delivered for 12 days beginning at the day of transplantation. Each animal was examined by operating microscopy. The grafts were evaluated clinically by means of Holland's scoring system and graft survival was assessed by Kaplan-Meier analysis. The neovascular indexes of rejection were compared among different groups. Histological examination on ocular tissues was performed on day 14 to confirm the clinical diagnosis of rejection. RESULTS: The average transplant survival time in the allogenic control (Group B) was (11.0 +/- 1.5) d. Treatment with RAPA (Group C) led to a statistically significant prolongation of transplant survival to (36.1 +/- 14.9) d (P < 0.05). Treatment with RAPA 3 mg.kg(-1).d(-1) prolonged transplant survival as compared with treatment with CsA (Group D), but the difference was not statistically significant (P > 0.05). Corneal neovascularization was induced after the surgery. In RAPA group, corneal neovascularization was markedly reduced as compared with allograft control (Group B) (P < 0.05) and CsA group (Group D) (P < 0.05). Fewer inflammatory cells were found in the corneal stroma of the RAPA group. CONCLUSION: These results show that oral immunosuppression with RAPA can prevent corneal graft rejection and corneal neovascularization. PMID- 16271189 TI - [Biopsy and submission of ophthalmic pathologic specimens]. PMID- 16271190 TI - [Clinical use of quantitative DNA pathologic examination in eye neoplasms]. AB - Quantitative DNA pathologic examination is an important content of quantitative pathology, which is to use cytochemistry quantitative analytical technique for quantitative analysis of patho-histological structure and intra-cellular DNA level. Quantitative determination of DNA in tumor cells as an objective quantitative index reflecting biological behavior of tumor cells and has an important significance for distinguishing precancerous lesions, borderline tumors, benign and malignant tumors; and for estimating the malignant degree and prognosis. This review introduces the study about DNA quantitative analysis used in clinical pathology of eye neoplasms. PMID- 16271192 TI - [The application of modified classification of the aortic dissection]. AB - OBJECTIVE: To determine the indication, optimal operative procedures, plan and the estimation of the prognosis depending on the subtype of aortic dissection defined by the extension and extent of dissection. METHODS: The outcome of 708 patients with aortic dissection between January 1994 and December 2004 was analyzed. Among them 477 patients suffered from Stanford type A aortic dissection were treated. Type A dissection can be classified into 3 subtypes based on the pathological change of the aortic root. Type A1 (No pathological change type): 212 patients underwent ascending aorta replacements; Type A2 (mild pathological change type): 63 patients underwent ascending aortic replacement with concomitant aortic valve and valsalva sinus plasty and David procedure was performed in 9 patients; Type A3 (severe pathological change type): 193 patients underwent Bentall procedure. The method of aortic arch repair was determined by the pathological type of distal aorta. Total aortic arch replacement was performed in 78 patients with complex type (type C). There hundred and ninety-nine patients with simple type (type S) underwent partial aortic arch replacement. 231 patients suffered from Stanford type B aortic dissection. Type B dissection can be classified into 3 subtypes based on dilated extension of proximal descending aorta. Type B1 (no dilation was confined in the proximal of thoracic descending aorta): endoluminal stent graft repair was performed in 103 patients. Replacement of the partial proximal thoracic descending aorta and replacement combined with stented elephant trunk procedure were performed in 32 and 12 patients respectively; Type B2 (aneurysm in thoracic descending aorta): 32 patients underwent the part proximal thoracic descending aorta replacement combined with aorta plasty. 21 patients underwent the replacement of entire thoracic descending aorta; Type B3 (aneurysm in thoracic descending and abdominal aorta): thoracoabdominal aortic replacement was operated in 31 patients with deep hypothermia circulatory arrest; Type BC (complex type): 44 patients were performed the operation with the use of deep hypothermia circulatory arrest because their left subclavian arteries or distal aortic arch were affected by the dissection; Type BS (simple type): 103 patients were underwent endoluminal stent graft repair. In the 60 patients, the operations were performed by using the technique which preserved blood was transfused back by pump via the femoral artery. Femoro-femoral bypass was performed in 24 patients. RESULTS: Type A: the operative mortality was 4.6% (27/477), and the hospital morbidity was 14.5% (69/477). Type B: the hospital mortality of endoluminal stent graft repair was 1.9% (2/103). 9.7% (10/103) had mild leakage from proximal communications. The morbidity was 2.9% (3/103) in stent group. The mortality was 3.1% (4/128), and the hospital morbidity was 18.8% (24/128) in the operative group. CONCLUSION: The subtype of aortic dissection is much useful in determining the optimal procedure, operative indication and plan, estimating the prognosis. PMID- 16271193 TI - [Surgical management for Stanford A aortic dissection]. AB - OBJECTIVE: To summarize the surgical experience for Stanford A aortic dissection. METHODS: Sixty-eight patients with Stanford A aortic dissection underwent surgery from March 1998 to October 2004, acute aortic dissection in 45 cases, chronic aortic dissection in 23 cases. The operation were performed by using moderate hypothermic cardiopulmonary bypass in 53 cases, deep hypothermic circulatory arrest (DHCA) and retrograde cerebral perfusion (RCP) in 11 cases; DHCA with antegrade selective cerebral perfusion (SCP) in 4 cases. Surgical procedures included ascending aortic grafting in 7 cases, ascending and hemiarch grafting in 6, ascending and total arch grafting in 3, ascending and total arch grafting with Frozen elephant trunk procedure in 4. Concomitant procedures included Bentall procedure in 34 cases, Wheat procedure in 12 cases, aortic valvuloplasty in 2 cases, mitral valvuloplasty in 1 cases. Urgent surgery was in 39 cases (emergency surgery in 19). RESULTS: Operative mortality was 7% (urgent surgery mortality was 8%, elective surgery mortality was 7%). Fifty-eight cases were followed up for (37 +/- 22) months. Actuarial survival of 58 cases at 1, 3 and 5 years was 100%, 95% and 86% respectively. CONCLUSION: The choice of surgical procedures depend on the location of intimal tear for Stanford A aortic dissection. Proper surgical indication, technique and brain protections are the key factors of Stanford A aortic dissection surgery. PMID- 16271194 TI - [Dexterously use four branch vessel prosthesis on aortic surgery]. AB - OBJECTIVE: To review the experience of various positions aortic replacement by four branch prosthesis vessel. METHODS: From August 2003 to May 2005, we finished aortic procedures with four branch prosthesis vessel for 142 patients, aged (44 +/- 12) (22-78) years, weighted (72 +/- 20) kg (49-130 kg). We performed ascending aorta and total aortic arch replacement for 85 cases during right axillary artery cannulation for cardiopulmonary bypass and selected antegrade cerebral perfusion. 38 patients underwent one-stage total thoracoabdominal aortic replacement during deep hypothermic bypass and subsection circulatory arrest. 8 patients underwent one-stage total or subtotal aortic replacement during deep hypothermic bypass and selected antegrade cerebral perfusion and subsection circulatory arrest. We performed totally aortic arch replacement without utilizing cardiopulmonary bypass and hypothermic for 11 cases. RESULTS: The mortality was 4.2%. Cerebral complications occurred in 16 (11.3%). 2 patients suffered from permanence spinal cord dysfunction. 4 patients suffered from temporary spinal cord dysfunction. CONCLUSION: The four branch vessel prosthesis can be used on aortic surgery dexterously. The approach may shorten she time of aortic arrest and arterial construction. PMID- 16271195 TI - [Application of endovascular thoracic branched aortic stent-grafts in the treatment of aortic arch dissection]. AB - OBJECTIVE: To report the initial clinical experience of endovascular thoracic branched stent grafts in the treatment of aortic arch dissections involving the left subclavian artery. METHODS: From February 2004 to June 2004, 14 patients were cured with the endovascular thoracic branched aortic stent-grafts made by Beijing YuHengJia SciTech Co. All patients had Stanford type B aortic dissection with the entry tears just beyond the origin of the left subclavian artery by an average distance of 8.7 mm. The branched stents were consisted of the aortic section and the branched section. The diameter of the stents was 15% to 20% larger than the diameter of the landing zones of native arteries. The repair procedure was performed in angiography laboratory. The branched stent grafts were delivered under fluoroscopic guidance and implanted into the aortic arch including the left subclavian artery. RESULTS: Fourteen branched stent-grafts and 2 additional flexible stent-grafts were delivered successfully in all 14 cases. The entry tears were excluded completely, and the truth lumen of the dissection was revealed to the normal diameter in all patients. Neither peripheral complication nor death occurred. All 14 patients had recover the normal life. CONCLUSION: It demonstrates that it is possible to apply the technical feasibility of endovascular thoracic branched aortic stent graft to repair the intimal tear of dissection just beyond the left subclavian artery. PMID- 16271196 TI - [The endovascular repair of ruptured aortic aneurysm]. AB - OBJECTIVE: To discuss the safety, feasibility, efficacy and problem of endovascular repair (EVR) for ruptured aortic aneurysm (RAA). METHODS: Fourteen patients with RAA were obtained diagnosis and anatomic data of proximal and distal neck through magnetic resonance imaging and CT. The patients with hypotension were accepted anti-shock treatment. Stent-grafts were deployed in proper position of RAA under X-ray fluoroscopic. RESULTS: The time from rupture to operation was 50 min to 21 d. Three cases had plenty of blood in left pleural cavity and 2 cases were done under bleeding shock condition. Five cases with RAA were fixed by bifurcated stent-grafts and others by straight stent-grafts. Follow up term was 1 month to 38 months. One with ruptured thoracic aortic aneurysm died 4 h later after operation, another one died of left thoracic cavity infection 9 months later, and others lived well without complication. CONCLUSION: The EVR is a safe, efficient and feasible method. It will be a potential alternative to treat RAA in future, but more suitable for RAA with better proximal and distal landing zones. PMID- 16271197 TI - [Endovascular stent-graft exclusion of giant and long-segment thoracic aortic aneurysms]. AB - OBJECTIVE: To discuss the availability of endovascular stent-graft exclusion in giant and long-segment thoracic aortic aneurysms. METHODS: Endovascular stent graft exclusion was performed on 3 male patients with giant and long-segment thoracic aortic aneurysm. Multiple stent-grafts were connected as a long stent graft to repair these thoracic aortic aneurysms, of which one with 28 cm in length and 7.3 cm in maximum diameter was excluded by 4 stent-grafts with different diameters and same length of 130 mm. The preliminary bypass between right common carotid artery and left common carotid artery or left common carotid artery and left subclavian artery was performed on 2 patients. RESULTS: Three patients underwent the operation successfully, and achieved the good results. There was no endoleak at the conjunction of stent-grafts in 2 cases, and a little endoleak in 1 case, which disappeared 1 year after operation. One case presented temporal cerebral ischemia, which disappeared after 1 month. CONCLUSIONS: The endovascular deployment with multiple stent-grafts connection is a safe, mini invasive and effective therapeutic method for giant and long-segment thoracic aortic aneurysm, but long-term outcomes will be gotten after the follow-up study. PMID- 16271200 TI - [Experimental study of differentiation of canine bone marrow mesenchymal stem cell into fibroblasts in vitro]. AB - OBJECTIVE: To explore the way of stably inducing canine bone marrow mesenchymal stem cells (BMSCs) to differentiate into fibroblasts and myofibroblasts in vitro, and provide seed cells for fabricating tissue engineering heart valves (TEHV). METHODS: Adult canine BMSCs were separated by a gradient centrifugation on Percoll (density 1.073 g/ml), then the cells were incubated in low-glucose Dulbecco Eagle's minimum essential medium (LG-DMEM) with 10% bovine calf serum. Cell phenotype were identified by immunohistochemistry staining. The second and third generation of BMSCs were committedly induced by conditioning culture medium, which were detected by immunohistochemistry staining. The induced-BMSCs were freezed, preserved and resuscitated after 7 d to observe the cell growth, proliferation and function. RESULTS: BMSCs deriving from the bone marrow mononuclear cells separated by a Percoll gradient were positive expression of alpha-smooth muscle antibody, vimentin and negative expression of CD34, laminin. About (50 +/- 3)% induced-BMSCs were positive expression of laminin. Approximately (85 +/- 3)% freezed induced-BMSCs could be resuscitated. And the growth, proliferation and function were well. CONCLUSION: BMSCs could be committedly induced to differentiate into fibroblasts and myofibroblasts in vitro. It is suitable to be the seed cells. PMID- 16271206 TI - [High-power holmium laser with percutaneous nephrolithotripsy for kidney calculi]. AB - OBJECTIVE: To assess the efficacy and safety of the high-power holmium laser with percutaneous nephrolithotripsy for kidney calculi. METHODS: The high-power (60 W: 3.0 J x 20 Hz) holmium laser with percutaneous nephrolithotripsy was performed on 52 patients with single kidney pelvic or calyceal stones (average stone diameter 3.1 cm), 36 patients with multi-kidney pelvic and calyceal stones (average diameter 2.8 cm), 24 patients with staghorn stones (average diameter 6.8 cm). The duration of stone surgery, stone-free rate and complication were assessed. RESULTS: The mean duration of stone surgery was 44 min, the stone-free rate was 66% (74/112) after the first session, 89% (100/112) at the end of session. 3 patients had high fever after percutaneous nephrolithotripsy, no other adverse events were noted. CONCLUSIONS: The high-power holmium laser with percutaneous nephrolithotripsy can fragments calculi quickly and reduces the length of time of operation. It is an effective and safe technique for kidney calculi. PMID- 16271207 TI - [Transperitoneal laparoscopic enucleation of renal angiomyolipoma: a report of 10 cases]. AB - OBJECTIVE: To evaluate the feasibility and clinical effect of transperitoneal laparoscopic enucleation of renal angiomyolipoma (RAML) without obstruction of renal pedicle. METHODS: Ten patients with renal angioleiomyoma (tumor diameter < 4 cm) were operated by transperitoneal laparoscopy without obstruction of renal pedicle. The operating time, blood loss, hospital stay after operation, intraoperative and postoperative complications and the operative effect were observed. RESULTS: All the 10 patients underwent the operation successfully. The average operating time was 90 min, average blood loss was 80 ml, the average hospital stay after operation was 7 d. No intraoperative or postoperative complications occurred. Follow-up period was 3-19 months and no tumor metastasized or occurred again. CONCLUSION: This mininvasive procedure is a more precise and complete method than before, which can minimize the blood loss and make patients recover quickly, so it is well worth clinical applying. PMID- 16271208 TI - [Heme oxygenase-1 gene transfer protects rat kidney transplant from ischemia/reperfusion injury]. AB - OBJECTIVE: To investigate the protective effect of Heme oxygenase-1 (HO-1) gene transfer on rat renal autograft against ischemia/reperfusion injury. METHODS: HO 1 recombinant adenovirus vectors were constructed and transduced into rat renal autograft by renal arterial perfusion. The renal autografts were transplanted orthotopically after store at 4 degrees C for 24 h, followed by contralateral native nephrectomy 5 d after transplantation. There were 25 rats in the control group. 5 h and 3 d after transplantation, reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemistry were used to detect the expression of HO-1 gene; enzyme-labeled immunosorbent (ELISA) was used to measure HO-1 protein content in the homogenate of renal autograft. RESULTS: The intensity of HO-1mRNA expression at 3 h and 3 d after transplantation were 0.65 +/- 0.11, 0.86 +/- 0.17 in the experimental group and 0.09 +/- 0.01, 0.15 +/- 0.02 in the control group respectively. The differences between the two groups were significant (t = 14.38, 11.73, P < 0.05). HO-1 protein content at 3 h and 3 d after transplantation were significantly increased in the experimental group, as compared with the control group [(297 +/- 61) ng/g and (468 +/- 51) ng/g versus (98 +/- 30) ng/g and (155 +/- 31) ng/g; t = 8.27, 14.83, P < 0.05]. HO-1 transduced autografts had less renal ischemic injury and lower serum creatinine level compared with control animals (P < 0.05). CONCLUSION: Adenoviral vector can successfully transduce rat kidneys with the HO-1cDNA, which can protect rat renal autografts from ischemia/reperfusion injury. PMID- 16271209 TI - [The dynamic effects of allogenic transplantations with the bladder acellular matrix grafts of rabbits]. AB - OBJECTIVE: To study the dynamic effects of allogenic transplantations with the bladder acellular matrix grafts (BAMG) of rabbits. METHODS: Hemi-cystectomies were performed in 25 rabbits, and the defects were repaired with BAMG about half bladder size. The rabbits underwent postoperative assessment of bladder function at 8 weeks, including cystometry, vesical volume, vesical compliance and cystography. The allografts were observed by light microscope and electron microscope at 1, 2, 4, 8, 12, 16 weeks after surgery. RESULTS: Macroscopic observation revealed that BAMG regenerated gradually. All urodynamic results of 8 weeks after surgery were not different statistically as compared with these of preoperation (P > 0.05). Cystography revealed that the morphous of bladder was recovered. Epithelialization and neovascularity occurred accompanied by infiltration of inflammatory cell at 1 week. Smooth muscle cell and stratified epithelium regenerated 2 weeks after grafting. Neural elements formed around smooth muscle bundles as early as 4 weeks. Each component regenerated on the frame of BAMG sequentially. After 16 weeks, it was difficult to delineate the junction between the host bladder and BAMG by histology. CONCLUSION: After allogenic transplantation with rabbits' BAMG, the constitution and function of the allografts regenerate completely and gradually on the frame of BAMG. PMID- 16271220 TI - [The significance of liver hanging maneuver in liver-splitting anterior approach for hepatectomy]. AB - OBJECTIVE: To analyze the feasibility of developing a tunnel between inferior vena cava (IVC) and caudate lobe before passing a tape through it, and to explore the significance of liver hanging maneuver in liver-splitting anterior approach for hepatectomy. METHODS: Blunt dissection was used to develop the tunnel before a tape was passed through. A hemostatic plate was placed on the surface of liver parenchyma if needed. In the procedure of hepatectomy, the tape was pulled up to create an interspace between liver parenchyma and IVC so that the IVC can be protected during transection. RESULTS: Liver hanging maneuver was performed successfully in 47 cases. There were no severe complications related to the procedure in these cases. The procedure was terminated in 1 case because of severe bleeding. CONCLUSIONS: 1. Liver hanging maneuver is feasible in terms of anatomy and technique. 2. With liver hanging maneuver, IVC can be protected safely and the intrahepatic vessels and ductal system at the transaction line can be exposed clearly. It also makes anterior approach for hepatectomy safer and easier. PMID- 16271221 TI - [Cyclosporine microemulsion C2 monitoring in Chinese adult liver transplant recipients: a preliminary randomized control trial]. AB - OBJECTIVE: To evaluate the safety and reliability of cyclosporine microemulsion (CsA-ME) C(2) monitoring and to determine the target level of C(2) in Chinese adult liver transplant recipients. METHODS: 53 Chinese adult liver transplant recipients were randomly divided into three groups (group C(0), n = 17; group high level C(2), n = 18; group low level C(2), n = 18). Blood chemistry reflecting heart, liver and renal function and CsA level were examined at certain interval during follow-up. The change of immune status and episodes of rejection were also observed closely. RESULTS: The group low level C(2) had the lowest CsA oral dose (2.51 +/- 0.37 mg/kg/d), and had significant difference compared with the other groups (P < 0.01). The best liver, heart and renal function was observed in group low level C(2). The CD(4)(+)/CD(8)(+) ratio of group low level C(2) was 1.04 +/- 0.68, which had no significant difference with C(0) group. The rejection incidence of the three groups had no significant difference. group low level C(2) had highest clinical benefit ratio (72.22%), while the clinical benefit of group high level C(2) is the lowest (11.11%). CONCLUSIONS: With rational target level, C(2) monitoring can show us the proper oral dose of CsA which can decrease the side effects remarkably without rejection episodes increasing. The target level of C(2) in Chinese adult liver transplant recipients might be: 600-800 ng/ml 1 to 6 months posttransplant, 400-600 ng/ml 7-12 months posttransplant. PMID- 16271222 TI - [Synergistic effect of bromocriptine combining tumor necrosis factor-alpha on reversing multidrug resistance in a nude mouse model of liver neoplasm]. AB - OBJECTIVE: To investigate synergistic effect of bromocriptine (BCT) combining tumor necrosis factor-alpha (TNF-alpha) on reversing multidrug resistance in a nude mouse model of liver neoplasm. METHODS: Human hepatocarcinoma cell line HepG(2) (HepG(2) group), drug resistant hepatocarcinoma cell line HepG(2)/adriamycin (HepG(2)/ADM group) and hepatocarcinoma cell line transfected with TNF-alpha gene HepG(2)/ADM/TNF (TNF group and BCT group) were injected into the liver of nude mice via orthotopic implantation to establish multidrug resistance model of liver neoplasm in vivo. All the mice were injected with 5 fluouracil + adriamycin + mitomycin in abdominal cavity for 7 d. The mice in BCT group was simultaneously given bromocriptine through gastric canal. Size and weight of the tumor were measured. Furthermore tumor histological character and growth of the nude mice was observed and its chemosensitivity was tested. MDR associated genes and proteins (MRP, LRP) of implanted tumors were detected by immunohistochemical staining and reverse transcriptive polymerase chain reaction (RT-PCR), and apoptosis rate of hepatocarcinoma cells was detected by TUNEL assay. RESULTS: The nude mouse model of each cell line was all inoculated successfully. The tumor growth rate and weight were significantly different among groups (P < 0.05). After chemotherapy tumor growth inhibition rate was higher in BCT group (67%) compared to ADM and TNF groups (P < 0.01), and similar to HepG(2) group (54%). MDR1 and LRP mRNA could be detected in all groups, but TNF-alpha was detected only in TNF-alpha and BCT groups. Furthermore, MDR1 and LRP protein expression of tumors in TNF-alpha and BCT groups was low similar to HepG(2) group. The apoptosis rate of hepatocarcinoma cells was much higher in BCT group than in other groups (P < 0.05) with TUNEL assay. CONCLUSIONS: TNF-alpha gene can down-regulate the MDR associated genes and proteins expression for example MDR1, LRP, and lower its tumorgenesis. Moreover, bromocriptine can enhance the susceptibility of HepG(2)/ADM cells to cytotoxic drugs. PMID- 16271223 TI - [The study on immunomodulation of donor mesenchymal stem cells on discordant liver xenotransplantation]. AB - OBJECTIVE: To observe the influence of guinea pig mesenchymal stem cells (MSCs) culture in vitro on the immunological status in heterogeneous recipient, and to evaluate the immunomodulation of MSCs to the immune rejection of liver xenotransplantation. METHODS: The mononuclear cell fraction obtained by centrifugation over Percoll was cultured in vitro. Cell surface epitopes were analyzed by flow cytometry technique, and MSCs were incubated at the addition of adipocyte induction media, and stained in Oil Red O. The immunoglobulin and complement in rat recipient serum were assayed after infusion of donor MSCs. Thirty guinea pigs and thirty Wistar rats were randomly divided into three groups. All rat recipients undertook cyclophosphamide (CTX) infusion. Group A was MSCs infusion group, group B was saline infusion group and group C was dexamethasone (DXM) infusion group. Orthotopic guinea pig to rat liver transplantation model was established. The survival time and the immunopathology of graft were observed. RESULTS: According to flow cytometry assay, MSCs were positive for CD44 and CD29, while negative for CD34 and CD45. Lipid droplets were found in the MSCs cytoplasm after being incubated in adipogenic inducation media. The concentrations of IgG, IgM, C3 at all time point after MSCs infusion were significantly decreased than before (P < 0.05), while the concentrations of IgA and C4 did not changed. The survival time of group A (431 +/- 27 min) was obviously longer than group B (148 +/- 16 min) and group C (141 +/- 22 min) (P < 0.01). The intense hyperacute rejection rapidly occurred of group B and C after blood re-perfusion. The severe immunopathological injury could be observed at the grafts of group B and C. However, the hyperacute rejection of group A occurred slightly. CONCLUSIONS: MSCs can be identified by virtue of cell morphology, membrane phenotype and differential potential. MSC may play a role in the immunosuppression in hyperacute rejection in the liver xenotransplantation. PMID- 16271224 TI - [Analysis of 618 cases of radical resection of rectal carcinoma]. AB - OBJECTIVE: To investigate and analyse curative effects of Miles operation and anal sphincter preserving operation for rectal carcinoma in 20 years. METHODS: From 1984 to 2004, 618 cases of rectal carcinoma that underwent radical resection including Miles operation and anal sphincter preserving procedures were analysed retrospectively each 10 years, earlier 10 years from 1984 to 1994, and later 10 years from 1994 to 2004. RESULTS: Among the 618 cases, 492 (79.6%) were followed up. The median of the follow-up time was 5.4 years. In the earlier 10 years, local recurrence rate of post operation was 6.9% (14/201), for Miles operation and anal sphincter preserving procedures the local recurrence rate was 6.7% and 7.1% respectively. In the later 10 years, the local recurrence rate was 5.1% (15/291), 4.8% for Miles operation, 5.2% for anal sphincter preserving procedures. With the procedure of canular anastomosis of colon and rectal mucosa, the local recurrence rate was 4.9%. Overall five-year survival rate was 64.7% (130/201) in the earlier 10 years, 66.3% (59/89) for Miles operation, 63.4% (71/112) for anal sphincter preserving procedures. In the later 10 years, the five-year survival rate was 68.0% (198/291) in all, for Miles operation 66.3% (55/83), for anal sphincter preserving procedures 68.7% (143/208). With the procedure of canular anastomosis of colon and rectal mucosa, the five-year survival rate was 71.3% (62/87). CONCLUSIONS: The operation for rectal cancer should be chosen individually according to locus, biological character, and clinical stages. Anal sphincter preserving procedures are performed increasingly, and they provide the same five-year survival rate as Miles operation does, and the patient's quality of life can be improved obviously. PMID- 16271225 TI - [Comparison of two surgical approach for cardiac cancer]. AB - OBJECTIVE: To explore the preferable surgical approach for cardiac cancer. METHODS: One hundred and sixty patients with cardiac cancer underwent operation in two surgical approaches (epigastrium and left chest). Analysis was conducted on lymph nodes resected, stump positive rate, radical resectability rate, perioperative mortality, surgical complication rate, postoperative hospital days, survival rate between the two groups. RESULTS: In the epigastrium group and the left chest group the average number of resected lymph nodes was 15.7 and 10.6, respectively, the upper stump positive rate was 5.0% and 1.2%, and there was significant difference between the two groups (P < 0.05). The lower stump positive rate, perioperative mortality, radical resectability rate, surgical complication rate, postoperative hospital days, survival rate of the two groups were 1.2%, 0.0%, 81.2%, 10.0%, 10 d, 53.7% and 1.2%, 1.2%, 83.7%, 11.2%, 12 d, 56.2%, respectively, and there was no significant difference between the two groups (P > 0.05). CONCLUSION: There was no difference on the radical respectability rate and 5-year survival rate between the two groups. Thus the surgical approaches for cardiac cancer should based on the location and extent of lesions and the state of patients. PMID- 16271226 TI - [The role of tissue factor in the invasion and metastasis of colorectal carcinoma cells]. AB - OBJECTIVE: To investigate the role of tissue factor (TF) in the invasion and hematogenous metastasis of human colorectal carcinoma cells. METHODS: The eukaryotic expression vectors pcDNA3.1/Zeo bearing either sense or antisense TFcDNA were transfected into HT-29 and LoVo cells by the way of lipofactamine 2000. TF proteins in transfected cells were detected by Western Blot. Then the transfected and un-transfected tumor cells were implanted into nude mice (Balb/c Nu/Nu) to produce primary tumor, lung metastasis and liver metastasis respectively. RESULTS: HT-29 and LoVo cells with sense-TFcDNA transfection showed increased TF expression compared with the cells without transfection, but the cells with antisense-TFcDNA transfection got the contrary change. The primary tumor growth and invasive range, lung metastasis and live metastasis all increased in sense transfectants but reduced in antisense transfectants. CONCLUSIONS: TF can increase the invasion and hematogenous metastatic ability of human colorectal carcinoma cells. PMID- 16271227 TI - [Follow-up study in endovascular therapy for the renal artery stenosis]. AB - OBJECTIVE: To evaluate the safety and efficacy of endovascular therapy for renal artery stenosis. METHODS: Percutaneous transluminal renal angioplasty with stent (PTRAS) was performed on 33 consecutive patients with severe renal artery stenosis who suffered from poorly controlled hypertension or renal dysfunction. They were subsequently underwent 7 to 49 months clinical follow up for the effect of the procedure on renal function, blood pressure control, mortality. RESULTS: Angiographic success was obtained in 32 (97.0%) of the 33 patients. The mortality was 18.2%. After PTRAS, two (6.1%) died of myocardial infarction within 4 months. Four (12.1%) patients with preoperative serum creatinine (Scr) > or = 177 micromol/L died of uraemia within 17 approximately 28 months. Twelve and twenty four months after the procedure, systolic and diastolic blood pressure of 26 (78.8%) cases with preoperative Scr < 177 micromol/L significantly decreased (P < 0.05), with less antihypertensive medications taken and satisfactory renal function. CONCLUSION: For patients without serious cardiorenal disease, PTRAS has a beneficial effect on blood pressure and renal function. For patients with serious cardiorenal disease or preoperative Scr > or = 177 micromol/L, the mortality is higher. PTRAS should be performed prudently. The preservation of renal function may be enhance by using renal protection device. PMID- 16271228 TI - [Primary experimental study on the construction of tissue engineering blood vessel]. AB - OBJECTIVE: To explore the primary experimental methods to construct tissue engineering blood vessel. METHODS: Using the collagen-chitosan to prefabricate the scaffolds with 3-dimensional structure, the proliferated human endothelial cells (ECs), smooth muscle cells (SMCs) and fibroblasts act as the seed cells. The cells were seeded to scaffolds in two-step method, and engineering tissue were matured by static culture or bioreactor culture. Extracellular matrix contents and the platelet aggregation were examined in engineering tissue, tissue engineering blood vessels were taken as patches to repair the man-made defaults on the rats aorta. RESULTS: The proliferated human ECs, SMCs and fibroblast can hold activity and act as seed cells. The prefabricated scaffolds, with excellently cell and tissue biocompatibility, can facilitated cells adherence and upgrowth, the cells quantities and extracellular matrix contents in engineered tissue are time dependent increase (P < 0.05). Platelet aggregation tests confirm the tissue engineering blood vessel have some anti-coagulability. Using the engineering tissue patch to repair the default, 6 aortas in 8 animal were patency till 10 days post-operation. CONCLUSIONS: The seeding cells can be seeded on the 3-dimensional collagen-chitosan scaffolds and matured, the tissue engineering blood vessel can be constructed primarily. PMID- 16271229 TI - [Therapeutic neovascularization with autologous bone marrow CD34+ cells transplantation in hindlimb ischemia]. AB - OBJECTIVE: To explore whether transplantation of autologous bone marrow stem cells might augment angiogenesis and collateral vessel formation in a canine model of hindlimb ischemia. METHODS: CD34(+) stem cells were centrifugation through Ficoll and an immune magnetic cell sorting system from bone marrow (20 ml) of canine (n = 5) and induced into endothelial cells with VEGF in vitro, and expression of von Willebrand factor. Bilateral hindlimb ischemia was surgically induced in canines and Dil fluorescence labeled autologous stem cells were transplanted into the ischemic tissues. RESULTS: Four weeks after transplantation, fluorescence microscopy revealed that transplanted cells were incorporated into the capillary network among preserved skeletal myocytes. The stem cells transplanted group had more angiographically detectable collateral vessels, a higher capillary density (12.0 +/- 2.8 vs. 5.0 +/- 1.6 per field; t = 4.17 P < 0.05) and a higher ABI (0.58 +/- 0.14 vs. 0.32 +/- 0.11; t = 2.95, P < 0.05). CONCLUSIONS: Direct local transplantation of autologous bone marrow CD34(+) stem cells seems to be a useful strategy for therapeutic neovascularization in ischemic tissues in adults, consistent with "therapeutic vasculogenesis." PMID- 16271231 TI - [Influence of aerosols on the expression of cyclin B1, cyclin C and proliferating cell nuclear antigen in wound tissue healing of burned rat models]. AB - OBJECTIVE: To investigate the influence of aerosols on the expression of cyclin B(1), cyclin C and proliferating cell nuclear antigen (PCNA) in wound tissue healing of burned rat models. METHODS: Sprague Dawley (SD) rats were inflicted as the deep partial thickness burn models. Rats were randomly divided into experimental group and control group. The experimental group were treated with aerosols. Samples were collected in 1 approximately 10 postburn days. Immunohistochemistry and image analysis methods were conducted to examine the expression of cyclin B(1), cyclin C and PCNA in both experimental and control groups. RESULTS: The expression of cyclin C in experimental group was detected in nucleus of skin basal cell on the second postburn day, increased evidently at the fifth days and sustained at high expression level up to the tenth days after treatment. The expression of cyclin C in experimental group was significantly higher than control group (P < 0.05). The expression of PCNA was first observed in skin basal cell nucleus and hair follicle cell nucleus in both experimental and control group on the third postburn day. The expression of PCNA increased evidently at the fifth days in experimental after treatment and that increased evidently at the seventh days in control group, which showed there were lots of active proliferation cell. And the difference of the expression of PCNA between experimental and control group was significant (P < 0.01). The expression of cyclin B(1) was detected in nucleus and cytoplasm of skin basal cell in both groups on the third postburn day, and no difference between the experimental and control group (P > 0.05). CONCLUSIONS: Aerosols can up-regulate the expression of cyclin C and PCNA in skin basal cell nucleus. Therefore the aerosols can accelerate wound tissue healing. PMID- 16271232 TI - [Role of p38 mitogen-activated protein kinase in the pathogenesis of stress ulcer]. AB - OBJECTIVE: To determine whether the activation of p38 mitogen-activated protein kinase (MAPK) is involved in the pathogenesis of stress ulcer. METHODS: Model of stress ulcer was established with the treatment of rats with water-immersion restraint (WIR) stress. Ulcer index (UI) was macroscopically evaluated as a parameter of gastric mucosal lesions. Expression of phospho- and pan-p38 in gastric mucosa was detected using Western blot analysis. Tumor necrosis factor alpha (TNF-alpha) and Interleukin 1beta (IL-1beta) gene expressions were analyzed by Northern blot analysis. As indicated in some experiments, rats were pretreated with intravenous injection of the specific p38 MAPK inhibitor CNI-1493 prior to WIR stress and then the changes of UI and TNF-alpha and IL-1beta mRNA expression were examined. RESULTS: The p38 MAPK was persistently activated in the gastric mucosa of rats with WIR stress, with maximal activation after 1 h of stress [(6.8 +/- 3.2) fold of baseline levels, P < 0.01]. Inhibition of p38 MAPK activation with CNI-1493 led to a marked decrease in UI in WIR stress rats. Similarly, the increased gene expression of proinflammatory cytokines TNF-alpha and IL-1beta in gastric mucosa induced by WIR stress were significantly diminished by p38 MAPK inhibition. CONCLUSION: p38 MAPK might have an important role in the pathogenesis of stress ulcer. PMID- 16271240 TI - [Total knee replacement in valgus knee]. AB - OBJECTIVE: To investigate the methods and clinical results of total knee replacement (TKA) in patients with valgus knee deformity. METHODS: Between January 1996 and August 2004, 87 TKAs were performed by means of medial parapatellar approach, standard osteotomy and only lateral soft tissue release with posterior stabilized implants on 74 patients (11 men and 63 women) with valgus deformity. The average age at the time of operation was 62.93 years (range, 26-80 years). Clinical and radiographic evaluations including range of motion (ROM), Knee Society Score System (KSS) and the tibial and femur angle (T-F angle) were performed at follow-up. RESULTS: After a mean follow-up of 33.8 months (range, 5 months-9 years), the average ROM improved from 91 degrees (range, 70 degrees-120 degrees) preoperatively to 112.4 degrees (range, 80 degrees-130 degrees) postoperatively, the average KSS improved from 22.7 points (0-48 points) preoperatively to 81.7 points (range, 71-93 points) postoperatively. The average function score improved from 26.5 points preoperatively to 86.3 points postoperatively, the average T-F angle was corrected from 21.59 degrees (range, 12 degrees-40 degrees) of valgus preoperatively to 8.7 degrees (0 degrees-11 degrees) of valgus postoperatively. One knee had lightly instability at follow-up, one knee with patellar dislocation preoperatively had subdislocation postoperatively, no other complication occurred. CONCLUSIONS: The techniques of medial parapatellar approach, standard osteotomy and only lateral soft tissue release with posterior stabilized implants can correct a fixed valgus deformity very successfully in patients undergoing primary total knee replacement, and provides excellent results. PMID- 16271242 TI - [Clinical experience of treating infection after total hip arthroplasty]. AB - OBJECTIVE: To present the experience in the management of the infected total hip arthroplasty (THA). METHODS: Forty-three cases (44 hips) of infection after THA were reviewed. Eighteen women and 25 men with a mean age of 54 years (range, 24 81 years) were studied. Follow-up ranged from 5 months to 16 years (mean, 3.2 years). Treatment strategies included: one-stage revision (n = 7), two-stage revision (n = 15), resection arthroplasty (n = 13), debridement and retain prosthesis (n = 5), and others (n = 3). RESULTS: Forty cases had positive culture result with 59 organisms including 19 staphylococcus epidermidis and 10 staphylococcus aureus. Thirty-six cases had been followed up, and no case had infection recurrence. Twenty-nine cases had postoperative Harris hip score averaged 78.5 (45-98). CONCLUSIONS: The diagnosis and treatment of infection after hip replacement is very difficult. The violence and antibiotic resistant rate of the organisms is high. Two-stage reconstruction of the infected hip is useful and effective because of higher eradication rate of the infection and good postoperative functional result. PMID- 16271241 TI - [Management of the bone defects in revision total hip arthroplasty]. AB - OBJECTIVE: To determine the efficiency and clinical outcome of impaction bone graft in revision total hip arthroplasty (THA). METHODS: Seventy-two revision THAs by using impaction bone graft in 48 patients with severe bone loss were gotten average 25 months follow-up. Harris hip scores and X-ray were used to evaluate the results. RESULTS: The mean Harris hip scores improved from 44.6 before operation to 87.4 after operation. The excellent and good results were 90.3%. No stem subsidence and cup migration were found. Femoral fracture rate was 4.2%, and dislocation rate was 1.4%. CONCLUSION: Impaction bone graft is an efficacious method to restore bone stock in revision THA, and using anatomic stem and freeze-drying cancellous bone allograft could also get good clinical result. PMID- 16271243 TI - [Clinical risk factors for deep vein thrombosis after total hip and knee arthroplasty]. AB - OBJECTIVE: To analyze the clinical risk factors for deep vein thrombosis (DVT) after total hip and knee arthroplasty in Chinese patients who received prophylactic treatment for DVT. METHODS: We evaluated 128 total hip arthroplasty (THA) and total knee arthroplasty (TKA) in 95 patients performed at our center from April 2004 to August 2004, which included 48 THAs in 43 patients and 80 TKAs in 52 patients. There were 27 men and 68 women with a mean age of 59.77 years (range, 23-78 years). All patients had been given low-molecular-weight heparin before operation and for 7-10 days post-operation to prevent DVT. Color Doppler ultrasonography was used to detect DVT of bilateral lower extremities in all patients before operation and at 7-10 days after operation. Nineteen clinical factors were examined preoperation and 7-10 days post-operation in order to analyze their influences on DVT formation after surgery. RESULTS: There were 45 patients who developed DVT after operation. The incidence of DVT in all patients was 47.4% (45/95) and the incidence of proximal DVT was 3.2%. There were more asymptomatic DVT (57.8%, 26/45) than symptomatic ones, and some patients without DVT (14%, 7/50) presented some of the DVT symptoms. Logistic regression analysis demonstrated a definite association of female, obesity (representative by BMI), cement usage and diagnosed RA with DVT with odds ratio of 10.008, 3.094, 8.887, and 0.194 respectively. Other clinical factors had no statistically significant association with DVT. CONCLUSIONS: Female, obesity, and cement usage were the risk factors for DVT after THA and TKA, and diagnosed RA was the protecting factors for DVT after THA and TKA. Other clinical factors such as age, OA, type of implant, monolateral or bilateral operation, duration of anesthesia, surgery and bandage usage for blood control, time for immobilization et al were not the risk factors for DVT. PMID- 16271244 TI - [The clinical comparison of microendoscopic lumbar diskectomy with and without preservation of the ligamentum flavum]. AB - OBJECTIVE: To discuss the surgical technique, indication and the clinical effects of the Microscope Endoscopic Tubular Retractor System (METRx) for microendoscopic lumbar diskectomy with the preservation of the ligamentum flavum. METHODS: Two hundred and eleven patients underwent single-segment METRx, with ligamentum flavum preservation in 65 patients (Group A), and without preservation in 146 patients (Group B). The two groups were compared clinically. All the patients suffered from low back pain and radicular syndrome to some extent, and the diagnosis was affirmed by CT and/or MRI. After exposed the interlaminar space regularly, the superior, inferior and lateral edge of the ligamentum flavum was released, the 3-sided dissociative ligament pacth was retracted medially during the spinal manipulation and restored anatomically after disc removal and the decompression of the nerve root. RESULTS: According to the results of two groups, ligamentum flavum preservation technique was feasible under endoscope and helpful in reducing the scar formation in the spinal canal. CONCLUSIONS: The ligamentum flavum preservation technique enable the surgeons to preserve the natural barrier. It is feasible to perform METRx, and it is helpful in reducing the epidural fibrosis. PMID- 16271245 TI - [The treatment of deep wound infection after posterior thoracic and lumbar instrumentation]. AB - OBJECTIVE: To study the characteristics and treatment of the deep wound infection after thoracic and lumbar instrumentation. METHODS: Thirty-six cases of deep wound infection after thoracic and lumbar instrumentation were retrospectively reviewed. There were acute deep wound infection in 14 cases and delayed infection in 22 cases. The patients with acute infection were treated with debridement and continuous irrigation and suction. Internal fixators were removed in 3 cases for repeated infection. The patients with delayed infection were treated with internal fixator removal, debridement and continuous irrigation and suction. RESULTS: At follow-up evaluation, only 1 patient had recurrence of infection because of his complicating vertebral osteomyelitis. The most common organisms cultured in acute deep wound infection were staphylococcus aureus and colibacillus while staphylococcus epidermis, micrococcus and diphtheria bacillus in delayed infection. The white cell count and ESR were elevated in the acute deep wound infection while only the ESR elevated and the white cell count remained normal in the delayed deep wound infection. CONCLUSIONS: There may be different between the acute and delayed deep wound infection's pathology. The internal fixator could be remained in the acute deep wound infection which need be removed in the delayed deep wound infection. PMID- 16271247 TI - [The etiological study, diagnose and therapy of hand special chronic infections in littoral area]. AB - OBJECTIVE: To explore the etiological factors of hand special chronic infections and their relationship with tuberculosis, and to give evidence for clinical diagnosis as well as treatments. METHODS: From 2002 to 2004 pathologic inspection, acid-fast stain, bacterial cultication, mycobacterial cultivation were performed in all 29 cases of hand special chronic infections. RESULTS: All cases showed granulomatous lesions in pathological appearance, 2 positive in acid fast stain, 12 positive in bacteria cultivation, and 1 nocardiosis, 1 staphylococcus epidermidis, 7 M.marinum, 1 M.tuberculosis, 1 M.fortuitum, 1 M.kansasii. CONCLUSIONS: Non-tuberculo-mycobacterium (NTM) especially M.marinum are far more important as the major factor than tuberculosis and other bacterial in hand special chronic infections. Bacteria cultivation should be routine examined for all cases. PMID- 16271246 TI - [Accuracy of CT-based navigation of pedicle screws implantation in the cervical spine compared with X-ray fluoroscopy technique]. AB - OBJECTIVE: To evaluate the feasibility and accuracy of cervical spine pedicle screw fixation assisted by X-ray fluoroscopy and CT-based navigation system. METHODS: 145 cervical pedicle screws were placed assisted by X-ray fluoroscopy and 159 cervical pedicle screws were placed assisted by CT-based navigation system. Screw positions were evaluated by postoperative CT scans or C-arm X-ray 3 dimensional reconstruction. RESULTS: In the computer- assisted group, 155 (97.5%) screws were found completely within their pedicles compared with 133 (91.7%) correctly placed screws in the X-ray assisted group (P < 0.05). The process of navigation investigated in twenty patients showed that the mean time for registration and surface matching was 3.5 (range from 2 to 8 minutes) minutes and the mean time for screw-marker insertion was 2 (range from 1 to 3.5 minutes) minutes with the mean position deviation 0.31 mm (0.12-0.56 mm) per screw. Only twice C-arm fluoroscopy were performed to verify the accuracy of one screw position intraoperatively. No severe complications like neurovascular lesion occurred in both groups. CONCLUSION: CT-based navigation system can increase the accuracy of cervical pedicle screw implantation significantly. PMID- 16271248 TI - [Experimental study on the osseointegration of nanophase hydroxyapatite biograde coated implants]. AB - OBJECTIVE: To study the osseointegration of the nanophase hydroxyapatite biograde coated implants and host bone. METHODS: Nanophase hydroxyapatite biograde-coated implants, hydroxyapatite biograde-coated implants and noncoated Ti-6Al-4V implants were inserted into both femoral of Beagle canines the tissue response, dynamic osteogensis and SEM were studied at 4, 8 and 12 weeks. RESULTS: The degradation of nanophase hydroxyapatite was slower than hydroxyapatite, dynamic osteogensis and the form of osteoblast were better than the control groups. CONCLUSION: The biological reaction of Nanophase hydroxyapatite biograde-coated implants is better than hydroxyapatite coated implant. PMID- 16271250 TI - [Research on chondrogenic differentiation and immunologic response of allogeneic mesenchymal stem cells implanted into joint cavity]. AB - OBJECTIVE: To evaluate the cartilage formation ability of allogeneic mesenchymal stem cells implanted into sheep joint cavity without the use of immunosuppressive therapy. METHODS: Allogeneic mesenchymal stem cells (MSCs) loaded onto porous beta-tricalcium phosphate ceramic (beta-TCP) were implanted into normal sheep joint cavity. A complete mismatch between donor stem cells and recipient sheep was confirmed by mixed lymphocyte reaction assays prior to implantation. Eight weeks after implantation, the implants were taken out for histological and immunohistochemical analysis. The histological results were compared with data derived from joint cavity implantation of autologous MSCs-ceramic composites and cell-free ceramics. The systemic immune response was evaluated by the analysis of recipient serum for production of antibodies against allogeneic cells. RESULTS: For implantation with allogeneic MSCs, no sign of adverse immune response was detected. Histologically, few inflammation cells infiltration occurred and no antibodies against allogeneic cells were detected. Neocartilage formation in implants loaded with either allogeneic or autologous mesenchymal stem cells was revealed by histochemical and immunohistochemical analysis. In implants without stem cells, no cartilage formation was detected. CONCLUSIONS: Allogeneic mesenchymal stem cells are capable of forming cartilage under the effect of joint cavity environment. Without the use of immunosuppressive therapy, allogeneic MSCs do not provoke an adverse immune response in vivo. PMID- 16271251 TI - [Chitin biological tube bridging the peripheral nerve with a small gap]. AB - OBJECTIVE: To investigate the possibility of bridging small peripheral nerve gap using a de-acetyl chitosan conduit. METHODS: The sciatic nerves of right sides were cut at SD rats. They were divided into 5 Groups randomly; Group A: epineurium suture in situ (n = 24); Group B: biological conduit with a small gap for bridging the peripheral nerve (n = 24, with 5 mm gap); Group C: epineurium suture with distal stump rotated 180 degrees (n = 24); Group D: bridging the nerve by biological conduits with a small gap, but the distal stump rotated 180 degrees (n = 24, with 5 mm gap); Group E: biological conduit with a small gap for bridging the peripheral nerve with NGF (n = 24). Electrophysiological examination, histological examination and myelinated axon counting were applied after 2, 4, 6, 8 weeks after operation respectively. RESULTS: Regenerated nerve fibers were seen in the distal nerve segments of all 5 groups; The nerve conduction velocity of small gap group (group B, D) was faster than that of corresponding simple epineurium suture group (group A, C) at all 2, 4, 6, 8 week time point (P < 0.05). The myelinated axon counting of small gap group (group B, D) was faster than that of corresponding simple epineurium suture group (group A, C) at all 4, 6, 8 week time point (P < 0.01), and there was no statistically significant difference at 2 week time point. CONCLUSION: The repair effects of chitin conduit bridging peripheral nerve with small gap (5 mm) are better than that of epineurium suture directly, and possess the potential to substitute the epineurium suture. PMID- 16271258 TI - The role of basal insulin in type 2 diabetes management. PMID- 16271252 TI - [The expression characteristics and biological significance of nuclear factor- kappa B in mice bone tissue of experimental osteoporosis models]. AB - OBJECTIVE: To study the expression characteristics and biological significance of nuclear factor NF-kappaB p50 and NF-kappaB p65 in osteoporosis development. Adult mice were ovariectomized as models of experimental osteoporosis. In this way, we can explore the molecular mechanism of osteoporosis and understand the significance during the morbility of osteoporosis. METHODS: Four-month female BALB/c mice were randomly divided into ovariectomized group and sham-operated group. 12 weeks after surgery, the mice were sacrificed after the measurement of bone mineral density (BMD). The NF-kappaB level in bone tissue were determined by immunohistochemistry and in situ hybridization techniques. RESULTS: Compared with the sham-operated group, bone density, rarefaction of trabecula and the number of osteoblast decreased in the ovariectomized group, while the number of osteoclast increased in the ovariectomized group. The immunohistochemistry showed that NF kappaB expressed in both groups. It mainly localized in the cytoplasm of osteoblast, osteocytes and marrow stroma cell. The expression level in the ovariectomized group was higher than that in the sham-operated group and was negatively correlated with the BMD (P < 0.01 or P < 0.05). In situ hybridization demonstrated that the expression levels of NF-kappaB 50mRNA and NF-kappaB 65mRNA in ovariectomized group were significantly higher than that of the sham-operated group (P < 0.05). CONCLUSION: The expression level of NF-kappaB significantly increased in the bone tissue of ovariectomized mice, and its abnormal expression was significantly correlated with BMD. PMID- 16271259 TI - Minimally invasive procedures: what family physicians need to know. PMID- 16271261 TI - Advances in fluorescence derivatization methods for high-performance liquid chromatographic analysis of glycoprotein carbohydrates. PMID- 16271262 TI - alpha-Lactalbumin binding and membrane integrity--effect of charge and degree of unsaturation of glycerophospholipids. AB - Several studies have shown that the physical state of the phospholipid membrane has an important role in protein-membrane interactions, involving both electrostatic and hydrophobic forces. We have investigated the influence of the interaction of the calcium-depleted, (apo)-conformation of bovine alpha lactalbumin (BLA) on the integrity of anionic glycerophospholipid vesicles by leakage experiments using fluorescence spectroscopy. The stability of the membranes was also studied by measuring surface tension/molecular area relationships with phospholipid monolayers. We show that the degree of unsaturation of the acyl chains and the proportion of charged phospholipid species in the membranes made of neutral and acidic glycerophospholipids are determinants for the association of BLA with liposomes and for the impermeability of the bilayer. Particularly, tighter packing counteracted interaction with BLA, while unsaturation-leading to looser packing-promoted interaction and leakage of contents. Equimolar mixtures of neutral and acidic glycerophospholipids were more permeable upon protein binding than pure acidic lipids. The effect of lipid structure on BLA-membrane interaction and bilayer integrity may throw new light on the membrane disrupting mechanism of a conformer of human alpha-lactalbumin (HAMLET) that induces death of tumour cells but not of normal cells. PMID- 16271263 TI - Inhibitory effect of DA-125, a new anthracyclin analog antitumor agent, on the invasion of human fibrosarcoma cells by down-regulating the matrix metalloproteinases. AB - Matrix metalloproteinases (MMPs), zinc-dependent proteolytic enzymes, play a pivotal role in tumor metastasis by cleavage of extracellular matrix as well as non-matrix substrates. In this study, we examined the influence of DA-125, a new anthracyclin analog, on the gene expression of MMPs (MMP-2, MMP-9 and MT1-MMP), tissue inhibitor of metalloproteinases (TIMP-1 and TIMP-2) and in vitro invasiveness of human fibrosarcoma cells. Dose-dependent decreases of MMPs and TIMPs mRNA levels were observed in DA-125-treated HT1080 human fibrosarcoma cells detected by reverse transcriptase-polymerase chain reaction. Gelatin zymography analysis also showed a significant down-regulation of MMP-2 and MMP-9 expression in HT1080 cells treated with DA-125 compared to controls. In addition, DA-125 inhibited the invasion, motility and cell migration, and colony formation of tumor cells. These data, therefore, provide direct evidence for the role of DA 125 as a potential cancer chemotherapeutic agent, which can markedly inhibit the invasive capacity of malignant cells. Further, to clarify the transcriptional regulatory pathway, we primarily investigated the role of nuclear factor-kappaB (NF-kappaB) in the expression of MMPs by DA-125 in HT1080 cells. Electrophoretic mobility shift assay demonstrated that DA-125 modulates the binding activity of NF-kappaB. Using the luciferase reporter gene assay, a dose-dependent down regulation of NF-kappaB-mediated luciferase expression was also observed. These results suggest that DA-125 down-regulates MMPs expression in HT1080 cells through the NF-kappaB-mediated pathway. PMID- 16271264 TI - Placental passage of tricyclic antidepressants. AB - BACKGROUND: The use of antidepressants during pregnancy continues to garner considerable attention, though there are limited investigations that have sought to quantify fetal exposure. METHODS: Maternal and umbilical cord sera were collected at delivery from ten women taking nortriptyline and seven taking clomipramine. Placental passage was calculated as the ratio of umbilical cord to maternal serum concentration. Obstetrical outcome data were gathered from subjects at delivery. RESULTS: The placental passage ratio of nortriptyline and its active metabolite, cis-10-hydroxynortriptyline, were .68 +/- .40, 1.40 +/- 2.40, respectively. Clomipramine and desmethylclomipramine ratios were .60 +/- .50, .80 +/- .60. Obstetrical complications, such as pre-term delivery and pregnancy induced hypertension, were increased compared to the national average. CONCLUSIONS: The in vivo ratios of umbilical cord to maternal serum drug concentrations demonstrate considerable fetal exposure and differ greatly from previous results utilizing ex vivo perfusion. PMID- 16271265 TI - Ectodermal Wnt-6 promotes Myf5-dependent avian limb myogenesis. AB - Limb muscles of vertebrates are derived from precursor cells that migrate from the lateral edge of the dermomyotome into the limb bud. Although several signaling molecules have been reported to be involved in the process of limb myogenesis, none of their activities has led to a consolidate idea about the limb myogenic pathway. Particularly, the role of ectodermal signals in limb myogenesis is still obscure. Here, we investigated the role of the ectoderm and ectodermal Wnt-6 during limb muscle development. We found that ectopic expression of Wnt-6 in the limb bud specifically extends the expression domains of Pax3, Paraxis, Myf5, Myogenin, Desmin and Myosin heavy chain (MyHC) but inhibits MyoD expression. Ectoderm removal results in a loss of expression of all of these myogenic markers. We show that Wnt-6 can compensate the absence of the ectoderm by rescuing the expression of Pax3, Paraxis, Myf5, Myogenin, Desmin and MyHC but not MyoD. These results show that, in chick, at least two signals from the limb ectoderm are necessary for muscle development. One of the signals is Wnt-6, which plays a unique role in promoting limb myogenesis via Pax3/Paraxis-Myf5, whereas the other putative signaling pathway involving MyoD expression is negatively regulated by Wnt-6 signaling. PMID- 16271266 TI - Pudendal-to-bladder reflex in chronic spinal-cord-injured cats. AB - The effects of pudendal nerve stimulation on reflex bladder activity were investigated in cats with chronic spinal cord injury (6-12 months) under alpha chloralose anesthesia. Electrical stimulation of the pudendal nerve on one side at different frequencies and intensities induced either inhibitory or excitatory effects on bladder activity. The inhibitory effect peaked at a stimulation frequency of 3 Hz and gradually decreased at lower or higher frequencies. The inhibitory effect could occur at stimulation intensities between 0.3 and 1 V (pulse width 0.1 ms) and increased at intensities up to 10 V. Stimulation of the central end of transected pudendal nerve also inhibited bladder activity, indicating that afferent axons in pudendal nerve are involved. Nerve transections also showed that both hypogastric and pelvic nerves might be involved in the inhibitory pudendal-to-bladder spinal reflex. Pudendal nerve stimulation at 20 Hz and at the same intensities (1-10 V) elicited a bladder excitatory response. Although this excitatory effect could not sustain a long lasting bladder contraction at small bladder volumes, it did induce continuous rhythmic bladder contractions at large bladder volumes. This study indicated the possibility of developing a neuroprosthetic device based on pudendal nerve electrical stimulation to restore micturition function after spinal cord injury. PMID- 16271267 TI - Depletion solid-phase microextraction for the evaluation of fiber-sample partition coefficients of pesticides. AB - A depletion solid-phase microextraction (SPME) method for the characterisation of SPME sorption for 13 pesticides selected as probe compounds is described. The sample is extracted and analysed multiple times by SPME-GC/MS. The observed depletion in peak areas is used for the calculation of extraction ratios that varied between 3 and 28% for a PDMS fiber with confidence intervals between 0.7 and 5.4%. Apparent fiber-sample partition coefficients can be calculated and extrapolated to equilibrium conditions if specific sorption kinetics are known. Under the chosen conditions, problems were encountered for more polar compounds (logK(ow)<3) due to inefficient extraction. The extracted amount was found to be the decisive parameter for depletion SPME and the extraction conditions therefore need to be adapted to the polarity of the analyte. The importance of the initial analyte concentration especially for mixed-mode fibers is demonstrated. Compared with conventional external calibration using liquid injection, depletion SPME eliminates uncertainties due to solvent effects during injection. Furthermore, it does neither require authentic reference compounds nor knowledge of the initial analyte concentration, and thus can even be used for unknowns. PMID- 16271268 TI - Identification of isomeric 4-nonylphenol structures by gas chromatography-tandem mass spectrometry combined with cluster analysis. AB - The endocrine-disrupting effect of 4-nonylphenols (NP) formed from industrial detergents such as nonylphenol polyethoxylates is well known today. The technical mixture contains a great variety of 4-iso-nonylphenol isomers having different endocrine-disrupting activities. Currently used gas chromatography-mass spectrometry (GC-MS) protocols allow the detection of about 20 peaks, mostly co eluting isomers. In the present study, Product Ion mass spectrometry obtained by ion trap technology enhanced the selectivity in NP detection resulting in improved gas chromatographic resolution as well as structure assignment of the isomers. The structure proposals of 4-nonylphenol isomers given were derived from GC-MS-MS data processed by multivariate statistics. The cluster analysis allowed the classification of NP due to common structural features that were reflected in the mass spectra. The fragmentation pathways of three reference NP isomers, 4-(1 ethyl-1,4-dimethylpentyl)phenol (NP1), 4-(1,1,5-trimethylhexyl)phenol (NP2) and 4 (1-ethyl-1,3-dimethylpentyl)phenol (NP3), were investigated in more detail. They served as model compounds to aid the interpretation of spectra from unknown NP isomers. Structures of two groups of isomers, characterized by alpha-ethyl as well as alpha-propyl substituents, could be proposed. PMID- 16271269 TI - Critical contribution of nonlinear chromatography to the understanding of retention mechanism in reversed-phase liquid chromatography. AB - The retention of most compounds in RPLC proceeds through a combination of several independent mechanisms. We review a series of recent studies made on the behavior of several commercial C18-bonded stationary phases and of the complex, mixed retention mechanisms that were observed in RPLC. These studies are essentially based on the acquisition of adsorption isotherm data, on the modeling, and on the interpretation of these data. Because linear chromatography deals only with the initial slope of the global, overall, or apparent isotherm, it is unable fully to describe the complete adsorption mechanism. It cannot even afford clues as to the existence of several overlaid retention mechanisms. More specifically, it cannot account for the consequences of the surface heterogeneity of the packing material. The acquisition of equilibrium data in a wide concentration range is required for this purpose. Frontal analysis (FA) of selected probes gives data that can be modeled into equilibrium isotherms of these probes and that can also be used to calculate their adsorption or affinity energy distribution (AED). The combination of these data, the detailed study of the best constants of the isotherm model, the determination of the influence of experimental parameters (e.g., buffer pH and pI, temperature) on the isotherm constants provide important clues regarding the heterogeneity of the adsorbent surface and the main properties of the adsorption mechanisms. The comparison of similar data obtained for the adsorption of neutral and ionizable compounds, treated with the same approach, and the investigation of the influence on the thermodynamics of phase equilibrium of the experimental conditions (temperature, average pressure, mobile phase composition, nature of the organic modifier, and, for ionizable compounds, of the ionic strength, the nature, the concentration of the buffer, and its pH) brings further information. This review provides original conclusions regarding retention mechanisms in RPLC. PMID- 16271270 TI - Keeping time: effects of focal frontal lesions. AB - This study examined the performance of 32 normal subjects and 39 patients with focal lesions of the frontal lobes on two simple timing tasks-responding in time with a tone that regularly repeated at a rate of once every 1.5s, and then maintaining the same regular response rhythm without any external stimulus. The hypothesis was that lesions to the right prefrontal cortex would disrupt timing performance. The two main findings were (1) an abnormally high variability in the timing performance (both self-timed and tone-timed) of patients with lesions to the right lateral frontal lobe, particularly involving Brodmann area 45 and subjacent regions of the basal ganglia; (2) an increase in the variability of timing performance as the task continued in patients with lesions to the superior medial regions of the frontal lobe. These findings indicate that the right lateral frontal lobe is crucially involved in the ongoing control of timed behavior, either because of its role in generating time intervals or in monitoring the passage of these intervals. In contrast, the superior medial regions of the frontal lobe are necessary to maintain consistent timing performance over prolonged periods of time. PMID- 16271271 TI - Modelling the impact of opportunistic screening on the sequelae and public healthcare costs of infection with Chlamydia trachomatis in Australian women. AB - OBJECTIVES: To describe the health outcomes and public healthcare costs of a single screening for Chlamydia trachomatis in Australian women aged 15-34 years. METHODS: A decision analytic model was used to determine the epidemiological estimates of prevalence and costs of C. trachomatis infection and its diagnosis, treatment and sequelae. RESULTS: We estimate that in any female population in Australia, with a Chlamydia prevalence rate of 5.7% or higher, a single screening examination for Chlamydia is cost saving for the public healthcare system. CONCLUSIONS: We found that opportunistic screening of high-risk populations is likely to be cost saving to the public healthcare system, although there is not sufficient evidence to support periodic population screening. As our model uses conservative epidemiological and public healthcare cost estimates, the health and financial impacts of C. trachomatis used in the model may be an underestimate of the true costs of infection. PMID- 16271272 TI - Assessments of pharmacokinetic drug interactions and tolerability of albendazole, praziquantel and ivermectin combinations. AB - The pharmacokinetic interactions and tolerability of albendazole, praziquantel and ivermectin combinations were assessed in 23 healthy Thai volunteers (12 males and 11 females). The study was an open, randomised, three-way crossover design in which each subject attended the study on three separate occasions (Phases I, II and III), of 4 d or 8 d each, with at least 1 or 2 weeks (but not longer than 2 months) between each phase. All subjects received the three study drug regimens as follows: regimen I, oral praziquantel (40 mg/kg body weight); regimen II, oral ivermectin (200 microg/kg body weight) given concurrently with an oral dose of albendazole (400 mg); and regimen III, oral ivermectin given concurrently with albendazole and praziquantel. All treatment regimens showed acceptable tolerability profiles. The incidence of overall drug-related adverse events was significantly higher following regimens I (12/23) and III (7/23) compared with that following regimen II (0/23). Six statistically significant changes in the pharmacokinetic parameters of albendazole sulphoxide (Cmax, AUC0-infinity, Vz/F, CL/F), praziquantel (Vz/F) and ivermectin (AUC0-infinity) were observed when the three drugs were given concurrently. However, based on US Food and Drug Administration criteria, these changes were not considered of clinical relevance. PMID- 16271273 TI - Dracaenogenins A and B, new spirostanols from the red resin of Dracaena cochinchinensis. AB - A 12(13-->14)abeospirostanol dracaenogenin A (1) and a spirostanol dracaenogenin B (2) were isolated from Chinese dragon's blood, the red resin of Dracaena cochinchinensis (Agavaceae). Their structures were established as (14S,25R)12(13- >14)abeospirosta-5,13(18)-diene-1beta,3beta,15alpha-triol (1) and (25R) spirost-5 ene-1beta,3beta,14alpha,15alpha-tetrol (2) by means of spectroscopic analysis, especially by 2D NMR spectra, and X-ray crystallographic analysis. Dracaenogenin A (1) is the first example of a 12(13-->14)abeospirostane spirostanoid found in nature. Its biogenesis from ruscogenin (3) through namogenin (4) and 2 was tentatively proposed. PMID- 16271274 TI - Diagnostic and therapeutic laser conization for cervical intraepithelial neoplasia. AB - OBJECTIVE: The clinical efficacy of conservative treatment using Nd-YAG laser technique for cervical intraepithelial neoplasia (CIN) was evaluated in a large series of CIN patients based on the long-term follow-up results. METHODS: We have treated 2107 women preoperatively diagnosed as having CIN by Nd-YAG contact laser conization with vaporization of the base. Their postoperative histologic findings and clinical outcomes were evaluated. RESULTS: The cone specimen was reported as showing that 1956 (92.8%) of 2107 cases had no CIN or CIN 1-3 and 151 (7.2%) cases had invasive diseases. 1956 cases without invasive diseases were followed up for 16 to 252 months. Incomplete excision occurred in 230 (12.3%) of 1874 patients with CIN lesion in the cone specimen, but failure rate (persistence or recurrence) was only 1.2%. 192 (83.5%) of 230 postoperative CIN patients with positive surgical margins showed no abnormal cytology or colposcopy during follow up period. Preoperative underdiagnosis of biopsy results compared with cytologic or colposcopic findings elevated the risk for incomplete excision and failure rate. CONCLUSION: The combination of laser excision and vaporization of the base was useful to detect unexpected invasive disease and revealed excellent therapeutic effects for CIN. Preoperative cytologic or colposcopic findings should be taken into account for the conservative treatment of CIN. PMID- 16271275 TI - Hypoxia blocks 11beta-hydroxysteroid dehydrogenase type 2 induction in human trophoblast cells during differentiation by a time-dependent mechanism that involves both translation and transcription. AB - The present study was undertaken to determine (1) if hypoxia-induced down regulation of placental 11beta-hydroxysteroid dehydrogenase type 2 (11beta-HSD2; encoded by HSD11B2 gene) activity and protein in human trophoblast cells during in vitro differentiation was mediated at the level of HSD11B2 gene transcription; and (2) whether the reduced placental 11beta-HSD2 in pregnancies complicated with fetal growth restriction (FGR) was a consequence of intrinsic abnormalities in trophoblast cells. Trophoblast cells were isolated from uncomplicated pregnancies and those complicated with FGR at term, and cultured for up to 72 h under normoxic (20% oxygen) or hypoxic (1% oxygen) conditions. Under normoxia, 11beta HSD2 activity and protein increased progressively over the 72 h culture period, which was accompanied by a corresponding rise in 11beta-HSD2 mRNA. As demonstrated previously, hypoxia blocked the increase in levels of both 11beta HSD2 activity and protein within the first 24h. In contrast, although hypoxia also prevented the rise in 11beta-HSD2 mRNA, it did not do so until 48 h. This time-dependent effect of hypoxia on placental 11beta-HSD2 activity/protein and mRNA suggests a dual mechanism of action whereby hypoxia may induce a rapid down regulation of 11beta-HSD2 protein synthesis, which occurs initially at the level of translation, and later extends to the level of transcription. Indeed, transient transfection studies demonstrated that hypoxia diminished HSD11B2 promoter activity. When trophoblast cells isolated from FGR placentas were cultured and allowed to differentiate under the same conditions, they not only exhibited a similar pattern of 11beta-HSD2 activity and mRNA expression but also responded to hypoxia similarly to those from normal placentas. This suggests that the reduced placental 11beta-HSD2 in FGR is not due to intrinsic abnormalities in trophoblast cells, but likely a result of extrinsic factors associated with FGR. PMID- 16271276 TI - Immunolocalization of myosin-V in the peribronchial, intrapulmonary peritracheal plexuses of the Wistar rat. AB - The location of myosin-V in whole mount preparations of the peritracheal and intrapulmonary peribronchial plexuses of Wistar rats has been shown by using an affinity-purified antibody specific to the medial tail domain of myosin-V. Myosin V immunostaining was intense in the peritracheal and intrapulmonary peribronchial plexuses, allowing the visualization of neuronal cell bodies and fibers. Knowledge of the cellular localization and function of this class of myosin is an important achievement, as it allows the study of these plexuses so as to clarify the importance of the complex mechanism responsible for the functioning of the airways. PMID- 16271277 TI - TATA-binding protein and TBP-associated factors during herpes simplex virus type 1 infection: localization at viral DNA replication sites. AB - Host RNA polymerase II (RNAP II) is responsible for viral transcription of the herpes simplex virus type 1 (HSV-1) genome and is relocalized to viral DNA replication compartments. Thus, we investigated whether TATA-binding protein (TBP) and TBP-associated factors (TAFs) are recruited to sites of viral transcription and replication and whether TBP/TAF expressions are influenced upon infection. The protein levels of TBP, hsTAF1/TAF(II)250, hsTAF4/TAF(II)135, and hsTAF5/TAF(II)100 were constant during the early phase of infection and started to decrease late during infection. Only for hsTAF7/TAF(II)55 we sometimes observed a decrease already at 4-8h postinfection (p.i.). Concomitantly with the relocalization of RNAP II, TBP and hsTAFs were redistributed to sites of viral DNA replication and transcription. In the absence of viral DNA replication TBP/hsTAFs were present in distinct nuclear dots, however, enlargement of the nuclear structures did not take place. Our results show that HSV-1 infection has no influence on the protein levels of TFIID components and leads to a redistribution of TBP and hsTAFs to prereplicative sites that enlarge to viral DNA replication compartments. PMID- 16271278 TI - Physical exercise prevents age-related decline in precursor cell activity in the mouse dentate gyrus. AB - Physical activity induces adult hippocampal neurogenesis. We here show that the acute up-regulating effect of voluntary wheel running on precursor cell proliferation decreases with continued exercise, but that continued exercise reduces the age-dependent decline in adult neurogenesis. Cell proliferation peaked at 3 days of running. After 32 days of exercise this response returned to baseline. Running-induced proliferation of transiently amplifying progenitor cells led to a consecutive increase in the number of more mature cells. Increasing age reduced adult neurogenesis at 9 months to 50% of the value at 6 weeks and to 17% at the age of 2 years. At both 1 and 2 years, precursor cell divisions remained inducible by physical activity. Exercise from 3 to 9 months of age significantly reduced the age-dependent decline in cell proliferation but (presumably in the absence of additional stimuli) did not maintain net neurogenesis at levels corresponding to a younger age. We propose that physical activity might contribute to successful aging by increasing the potential for neurogenesis represented by the pool of proliferating precursor cells. PMID- 16271279 TI - Effects of phosphorus fertilization on the availability and uptake of uranium and nutrients by plants grown on soil derived from uranium mining debris. AB - Subterranean clover and barley were grown on a soil derived from uranium mining debris and fertilized with phosphate as a U immobilizing additive for in situ remediation. We investigated the beneficial effect of P fertilization in the range 0-500 mg P kg(-1) soil in terms of U extractability, plant biomass production and U uptake. Increasing P in the mining debris caused a significant decrease of the water-soluble U and NH(4)-Ac extractable U at pH 7 and 5. For both plant species, P fertilization considerably increased root and shoot dry matter up to a maximum observed for soil receiving 100 mg P kg(-1) while the soil to-plant transfer of U was regularly decreased by increasing P content in soil. These observations show that P fertilization represents an in situ practical option to facilitate the revegetation of U-mining heaps and to reduce the risks of biota exposure to U contamination. PMID- 16271280 TI - Allopurinol suppresses 2-bromoethylamine and 1-methyl-4-phenylpyridinium ion (MPP(+))-induced hydroxyl radical generation in rat striatum. AB - The present study was examined whether or not 2-bromoethyamine, a semicarbazide sensitive amine oxidase (SSAO, EC; 1.4.3.6) inhibitor, would increase an active dopaminergic neurotoxin, 1-methyl-4-phenylpyridinium ion (MPP(+))-induced hydroxyl radical ((*)OH) generation in the rat striatum. Rats were anesthetized, and sodium salicylate (0.5 mM or 0.5 nmol/microl/min) was infused through a microdialysis probe to detect the generation of (*)OH as reflected by the non enzymatic formation of 2,3-dihydroxybenzoic acid (DHBA) in the striatum. Infusion of 2-bromoethylamine (100 microM or 100 pmol/microl/min) into the striatum drastically increased the formation of (*)OH products, trapped as DHBA by the action of MPP(+). Further, I studied the effect of allopurinol, a xanthine oxidase inhibitor, an 2-bromoethylamine and MPP(+)-induced (*)OH generation. Allopurinol (10 microM or 10 pmol/microl/min) significantly suppressed 2 bromoethyamine and MPP(+)-induced (*)OH. These results suggest that a definite mechanism is not clear at the moment, after inhibition of tissue-bound and/or blood plasma SSAO activity, with consequent increases in bioactive amine levels, enhances the formation of (*)OH products of efflux/oxidation due to MPP(+). PMID- 16271281 TI - Endocrine disrupters: a human risk? AB - Endocrine disrupters (EDs) alter normal hormonal regulation and may be naturally occurring or environmental contaminants. Classically, EDs act genomically, with agonistic or antagonistic effects on steroid receptors and may alter reproductive function and/or cause feminisation by binding to oestrogen or androgen receptors; their binding to the thyroid receptor may dysregulate the neuroendocrine system. Recently, it has been shown that EDs can also act by non-genomic mechanisms, altering steroid synthesis (inhibition of cytochrome P450 isoforms) or steroid metabolism. The alkylphenol and phthalate plasticisers inhibit the inactivation of oestrogens by sulphation (via SULT 1A1 and 1E1 isoforms) and so cause a rise in levels of the free active endogenous oestrogens. A range of ED effects have been shown in mammals, fish, birds, reptiles, amphibia and aquatic invertebrates but it is not yet clear whether these processes also occur in human beings. It is evident that EDs, as well as altering reproduction, can cause changes in neurosteroid levels and so have the potential to affect immune function, behaviour and memory. This may be of long-term concern since traces of EDs such as plasticisers, brominated fire retardants, sunscreen agents and cosmetic ingredients are widely distributed in the environment and in human biofluids. PMID- 16271282 TI - Enhancement of pro-oxidant effect of 7,12-dimethylbenz (a) anthracene (DMBA) in rats by pre-exposure to restraint stress. AB - The current study was designed to assess the effect of immobilization stress on liver toxicity induced by topical as well as oral administration of 7,12-dimethyl benz(a)anthracene (DMBA) in Swiss Albino rats. The experimental animals were divided into six groups. Group 1 animals were exposed to chronic restraint stress alone for 10 days (3h/day), shaved back of animals in group II were painted with 0.5% solution of DMBA twice a week for 4 weeks. Group III animals were first exposed to restraint stress similar to group I followed by DMBA application as in group II, group IV animals were orally administered four doses of 0.5% DMBA solution. (1ml/rat) at weekly intervals, while group V animals were first exposed to restraint stress as in group I followed by oral dose of DMBA similar to group IV. The untreated Group VI animals served as controls. Rats were sacrificed after a period of 4 weeks following DMBA administration. Biochemical measurements were carried out on liver tissues and serum/plasma of control and treated animals. Restraint stress was found to have marked effect on DMBA induced alteration of liver function as revealed by the increase in tissue marker enzymes viz glutamate oxaloacetate transaminase (GOT), glutamate pyruvate transaminase (GPT), alkaline phosphatase (ALP), acid phosphatase (ACP), lactate dehydrogenase (LDH) with a significant further decrease in antioxidant enzymes catalase (CAT), superoxide dismutase (SOD), glutathione-S-transferase (GST), glutathione reductase (GR) as compared to controls and DMBA alone(topical/oral) or stress alone treated rats. Increased lipid peroxidation was accompanied by a significant decrease in the level of total reduced glutathione (GSH). The changes in the levels of marker enzymes and in vivo antioxidants in serum/plasma were comparable to that of liver. The results of the present study indicate that immobilization stress markedly enhances DMBA induced alteration of liver and circulatory antioxidant status of the rats irrespective of the mode of DMBA administration though with a predominant effect on orally infused DMBA. PMID- 16271283 TI - Solidification/stabilization of arsenic containing solid wastes using portland cement, fly ash and polymeric materials. AB - Stabilization/solidification (S/S) is used as a pre-landfill waste treatment technology that aims to make hazardous industrial wastes safe for disposal. Cement-based solidification/stabilization technology is widely used because it offer assurance of chemical stabilization of many contaminants and produce a stable form of waste. The leaching behavior of arsenic from a solidified/stabilized waste was studied to obtain information about their potential environmental risk. Activated alumina (AA) contaminated with arsenic was used as a waste, which was stabilized/solidified (S/S) using ordinary portland cement (C), fly ash (FA), calcium hydroxide (CH) and various polymeric materials such as polystyrene and polymethyl methacrylate (PMMA). Toxicity characteristics leaching procedure (TCLP) and semi-dynamic leach tests were conducted to evaluate the leaching behavior of arsenic. Formations of calcite along with precipitate formation of calcium arsenite were found to be responsible for low leaching of arsenic from the stabilized/solidified samples. Effective diffusivity of arsenic ion from the matrix and leachablity index was also estimated. Minimum leaching of the contaminant was observed in matrix having AA+C+FA+CH due to the formation of calcite. PMID- 16271284 TI - Antimicrobial action of novel chitin derivative. AB - Aminoethyl-chitin (AEC) was synthesized in an attempt to both increase solubility of chitin in water and biological activity. AEC was obtained by grafting 2 chloroethylamino hydrochloride onto chitin at C-6 position. The structure of AEC was elucidated FT-IR and (1)H NMR spectroscopy, and its antimicrobial activity was investigated using three Gram-positive and Gram-negative bacteria. The integrity of the cell membranes of the representatives E. coli and S. aureus was investigated by determining the release of intracellular components of cells. When treated with AEC, release of 260 nm absorbing materials quickly increased both E. coli and S. aureus, but absorbance value was different due to the difference in cell structures. For detailed study, outer membrane (OM) and inner membrane (IM) permeabilization assay were performed using the fluorescent probe 1 N-phenylnaphthylamine (NPN) and the release of cytoplasmic beta-galactosidase activity. The results showed that AEC rapidly increased NPN uptake and the release of cytoplasmic beta-galactosidase via increasing the permeability of OM and IM. In addition, cytotoxic effect of AEC was assessed using human lung fibroblast (MRC-5) cell line, and AEC showed less toxic against MRC-5. PMID- 16271285 TI - Genetic study of gestation length in Andalusian and Arabian mares. AB - The length of gestation in Andalusian, or Spanish Purebred (SPB) and Arabian (AB) mares reared in Spain was analysed, based on 766 spontaneous full-term deliveries appertaining to 141 mares of SPB breed and 72 mares of AB breed in 31 breeding seasons. The data were obtained from the Yeguada Militar de Jerez de la Frontera stud farm in Cadiz, Spain. The mean length of gestation was of 336.8+/-0.48 days in the SPB mares and 340.3+/-0.63 days in AB mares. To assess the accurate prediction of time of birth the potential effect of a number of factors was investigated. The influences of the breed, mare, month and year of mating, age of the mother, number of births and sex of the foal were statistically significant. The factor have the greatest influence over the gestation length was the mare itself, with a correlation among consecutive births of around 0.4. The effect of inbreeding, both of the mare and foal, was negligible. Gestation length shortened as the breeding season progressed: in both breeds, a delay of 1 month in mating corresponded to a decrease of 3 days in the gestation length. According to our results, gestation length decrease as the mare gets older, with the shortest gestation periods when the mare is 10-12 years old, and from this point on, it slowly increases. The gestation period shortens as the 4th or 5th birth approaches, and then gets progressively longer. The range of variation in gestation length due to the number of births to the mare is of 2.9 days for the AB mares, and 2.2 days for SPB mares. The heritability for the gestation length for AB and the SPB breeds was 0.2, with a repeatability of 0.36 and 0.37, for SPB and AB breeds, respectively. With the data from both breeds, and using a classical approach, the response to selection was estimated if mares with extreme gestation lengths were culled, i.e. lengths which are under 310 days, or over 360 days. According to our results, in the case of SPB, a decrease of 14-45% would occur in the number of extreme gestation lengths, while in the AB breed, this value would decrease from 2 to 39%. PMID- 16271286 TI - Integrated approaches towards drug development from Ayurveda and other Indian system of medicines. AB - Biodiversity of natural resources has served not only for the primary human needs but also for health care, since time immemorial. The Indian subcontinent, with the history of one of the oldest civilization, harbors many traditional health care systems. Their development was supported by the diverse biodiversity in flora and fauna due to variations in geographical landscaping. Ayurveda, whose history goes back to 5000 b.c., is one of the ancient health care systems. The Ayurveda was developed through daily life experiences with the mutual relationship between mankind and nature. The ancient text of Ayurveda reports more than 2000 plant species for their therapeutic potentials. Besides Ayurveda, other traditional and folklore systems of health care were developed in the different time periods in Indian subcontinent, where more than 7500 plant species were used. According to a WHO estimate, about 80% of the world population relies on traditional systems of medicines for primary health care, where plants form the dominant component over other natural resources. Renewed interest of developing as well as developed countries in the natural resources has opened new horizons for the exploration of natural sources with the perspectives of safety and efficacy. The development of these traditional systems of medicines with the perspectives of safety, efficacy and quality will help not only to preserve this traditional heritage but also to rationalize the use of natural products in the health care. Until recent past, the nature was considered as a compendium for templates of new chemical entities (NCEs). The plant species mentioned in the ancient texts of these Ayurveda and other Indian systems of medicines may be explored with the modern scientific approaches for better leads in the health care. PMID- 16271287 TI - Antibacterial and antifungal activity of traditional medicinal plants used against venereal diseases in South Africa. AB - Aqueous, ethanolic and ethyl acetate extracts of 13 plants used in South Africa for the treatment of venereal diseases were screened for antibacterial and antifungal activity. Among the plants tested, Gunnera perpensa, Harpephyllum caffrum, Hypoxis latifolia and Ledebouria ovatifolia showed the best antibacterial activity. The aqueous extracts of Gunnera perpensa and Harpephyllum caffrum were most active against all the tested bacteria. In antifungal screening, good activity was shown by the ethanolic extracts of Bersama lucens and Harpephyllum caffrum. Only in the case of Harpephyllum caffrum did aqueous extracts have activity against Candida albicans. PMID- 16271288 TI - Elemental analysis of soil samples for forensic purposes by inductively coupled plasma spectrometry--precision considerations. AB - Major and trace elemental composition provides a powerful basis for forensic comparison of soils, sediments and rocks. However, it is important that the potential 'errors' associated with the procedures are fully understood and quantified, and that standard protocols are applied for sample preparation and analysis. This paper describes such a standard procedure and reports results both for instrumental measurement precision (repeatability) and overall 'method' precision (reproducibility). Results obtained both for certified reference materials and example soils show that the instrumental measurement precision (defined by the coefficient of variation, CV) for most elements is better than 2 3%. When different solutions were prepared from the same sample powder, and from different sub-sample powders prepared from the same parent sample, the CV increased to c. 5-6% for many elements. The largest variation was found in results for certified reference materials generated from 23 instrument runs over an 18 month period (mean CV=c. 11%). Some elements were more variable than others. W was found to be the most variable and the elements V, Cr, Co, Cu, Ni and Pb also showed higher than average variability. SiO2, CaO, Al2O3 and Fe2O3, Rb, Sr, La, Ce, Nd and Sm generally showed lower than average variability, and therefore provided the most reliable basis for inter-sample comparison. It is recommended that, whenever possible, samples relating to the same investigation should be analysed in the same sample run, or at least sequential runs. PMID- 16271289 TI - Self-inflicted injuries with negative political overtones. AB - The report deals with two adolescents (a 17-year-old Turkish boy and a 15-year old German girl) who simulated right-wing assaults by cutting National Socialist symbols into their own skin. The fact that Nazi symbols were chosen was obviously due to the idea that the alleged commission of the offense by right-wingers would not be doubted due to the negative image of this group. The alleged victims did not inform the police until the family or close friends urged them to report the incident. The rapid elucidation of the facts was possible because medicolegal experts were called in at an early stage of the investigations. The injuries showed typical morphological features of self-infliction (localization in easily accessible parts of the body, multitude of singular lesions with a linear or slightly curved course, equally shallow and non-penetrating cuts, arrangement in groups or over a defined area, lack of defense injuries, no corresponding damage to the clothing). PMID- 16271290 TI - Array CGH using whole genome amplification of fresh-frozen and formalin-fixed, paraffin-embedded tumor DNA. AB - The ability to utilize formalin-fixed, paraffin-embedded (FFPE) archival specimens reliably for high-resolution molecular genetic analysis would be of immense practical application in the study of human disease. We have evaluated the ability of the GenomePlex whole genome amplification (WGA) kit to amplify frozen and FFPE tissue for use in array CGH (aCGH). GenomePlex gave highly representative data compared with unamplified controls both from frozen material (Pearson's R(2) = 0.898) and from FFPE (R(2) = 0.883). Artifactual amplification observed using DOP-PCR at chromosomes 1p, 3, 13q, and 16p was not seen with GenomePlex. Highly reproducible aCGH profiles were obtained using as little as 5 ng starting material from FFPE (R(2) = 0.918). This WGA method should readily lend itself to the determination of DNA copy number alterations from small fresh frozen and FFPE clinical tumor specimens, although some care must be taken to optimize the DNA extraction procedure. PMID- 16271291 TI - Genome-wide single nucleotide polymorphism analyses of rheumatoid arthritis. AB - Because of the limitations of candidate gene studies and linkage analyses for common diseases, genome-wide association studies are now recognized as a powerful approach to mapping responsible genes with modest effects on various diseases. We performed whole genome case-control linkage disequilibrium (LD) mapping for rheumatoid arthritis (RA)-associated genes in Japanese subjects using single nucleotide polymorphisms (SNPs) mainly discovered in gene-containing regions. We identified RA-associated polymorphisms in two genes/loci, PADI4 and SLC22A4/A5 cluster. PADI4 catalyzes the conversion of arginine residues to citrulline in proteins. Recent reports on the high specificity of autoantibodies against citrullinated proteins to RA and the results of our study suggest that citrullination by PADI4 is a fundamental phenomenon of RA. On the other hand, the functions of SLC22A4/A5 have not been studied in detail, but SLC22A4/A5 have been reported to have multiple polymorphisms associated with several autoimmune diseases. Thus, large-scale LD mapping appears to be effective for identifying RA associated polymorphisms. PMID- 16271292 TI - Increased levels of bioactive IL-16 correlate with disease activity during relapsing experimental autoimmune encephalomyelitis (EAE). AB - Experimental autoimmune encephalomyelitis (EAE) is a CD4+ T-cell mediated disease, which resembles immunopathology of multiple sclerosis (MS). Interleukin (IL)-16 is a CD4+ cell-specific chemoattractant cytokine. In CD4+ T cells, production of bioactive IL-16 from constitutive pro-IL-16 requires cleavage by active caspase-3. We reported reversal of established relapsing disease by IL-16 neutralization. To better understand role(s) of IL-16 in regulation of relapsing EAE, we comparatively analyzed levels of IL-16, active caspase-3 and CD4 in mice with severe relapsing-remitting [(B6xSJL) F1], and low-relapsing (B6), disease. Elevated levels of IL-16 along with an increase in active-caspase-3 and CD4 levels correlated with stages of clinically active disease in both strains. CNS levels of bioactive IL-16 were notably higher in F1 compared to B6 mice at all stages, being most prominent during relapse. Similar patterns of regulation for IL-16 and active caspase-3 were observed in peripheral lymphoid organs, and in T cells isolated from lymph nodes following T-cell activation in vitro. IL-16 was co-immunoprecipitated with CD4 from CNS of relapsing mice. Our data suggest that caspase-3 mediated production of IL-16 by infiltrating CD4+ T cells, contributes to ongoing neuroinflammation by chemoattraction of additional waves of CD4+ T cells. PMID- 16271293 TI - Synthesis and SAR of highly potent and selective dopamine D(3)-receptor antagonists: Quinolin(di)one and benzazepin(di)one derivatives. herve.geneste@abbott.com. AB - The synthesis and SAR of novel and selective dopamine D(3)-receptor antagonists based on a 3,4-dihydro-1H-quinolin-2-one, a 1,3,4,5-tetrahydro-benzo[b]azepin-2 one, 1H-quinoline-2,4-dione or a 3,4-dihydro-1H-benzo[b]azepine-2,5-dione scaffold are discussed. A706149 (2.15mg/kg, po) antagonizes PD 128907-induced huddling deficits in rat, a social interaction paradigm. PMID- 16271294 TI - Nurse-led discharge to the ward from high dependency: a service improvement project. AB - Nurse-led discharge from the High Dependency Unit (HDU) was introduced in 2002 with the aim of improving access to level 2 beds (beds offering higher level of support and observation; ). The project has yet to be formally evaluated but there have been perceived benefits for both patients and staff. PMID- 16271295 TI - Promoting care for acutely ill children-development and evaluation of a paediatric early warning tool. AB - The primary purpose of this paper was to develop and evaluate a physiologically based system for the identification of acutely ill children in hospital environments. The dependency of children in hospital is increasing and ensuring the appropriate and timely intervention by a team of health personnel experienced in the care of these children is paramount to ensure their optimal outcome. A paediatric early warning (PEW) tool was designed and demographic and physiological data collected on all children (n = 360) who triggered the tool over a 6-month period, between September 2003 and February 2004. Analysis of the data was undertaken on each criterion within the tool and by reviewing it against patient outcome, the decision for its retention or removal was made. The modified tool showed a 99% sensitivity and a 66% specificity. The resultant Paediatric Early Warning Tool has been validated for use in a tertiary children's hospital in the United Kingdom (UK). The use of such a tool by all staff caring for acutely ill children in hospital environments can help to ensure their early recognition and timely treatment. The tool together with an action plan must, however, be appropriate for use in individual ward or hospital areas. PMID- 16271297 TI - Social integration and concentrations of C-reactive protein among US adults. AB - PURPOSE: This study tests whether social integration is associated with C reactive protein (CRP) level, a biologic risk factor for cardiovascular disease. METHODS: Using data from 14,818 participants aged>or=20 years from the Third National Health and Nutrition Examination Survey (1988 to 1994), we created a social network index using marital status; number of contacts with family, friends, and neighbors; frequency of religious service attendance; and participation in voluntary organizations. Serum CRP concentration was measured by means of latex-enhanced nephelometry, a low-sensitivity assay, and dichotomized into 3 mg/L or less and greater than 3 mg/L. RESULTS: After adjustment for multiple potential confounders, men aged>or=60 years with the fewest ties were more likely to have an elevated CRP concentration than men with the most ties (odds ratio=1.80; 95% confidence interval, 1.11-2.92). This occurred in a dose response manner, with each decrease in number of ties associated with an increase in the proportion of men with elevated CRP levels. The association between social networks and CRP level after multivariate adjustment was not significant in women or younger men. CONCLUSION: In this nationally representative cohort, CRP level was associated with social integration in older men, but not women or younger men. There may be sex- and age-related differences in biologic processes influenced by social integration. PMID- 16271298 TI - Neural correlates of control processes engaged before and during recovery of information from episodic memory. AB - Preparing to retrieve episodic information engages content-specific pre-retrieval processes--retrieval orientations--that vary according to the episodic information that is to be retrieved [Rugg, M.D., Wilding, E.L., 2000. Retrieval processing and episodic memory. Trends Cogn. Sci. 4, 108-115]. Retrieval orientations are retrieval sets which can be maintained tonically and which influence the ways in which subsequent stimuli are processed, presumably in order to facilitate recovery of the required episodic information. Indices of processes related to the adoption of a retrieval orientation can therefore be obtained by acquiring measures of neural activity during preparation to retrieve, while processes contingent upon the successful adoption of an orientation can be obtained by measuring the neural activity that is elicited by stimuli to which memory judgments are required. Across three experiments, electrophysiological indices of the former class of process were obtained only when frequent switches between different retrieval tasks were required, confirming that this measure of brain activity reflects processes important for the adoption of a relevant retrieval orientation. Stimulus-specific indices of retrieval orientations were obtained primarily when few switches between retrieval tasks were required, consistent with the view that the engagement of stimulus-specific retrieval processing operations is achieved only when an appropriate retrieval orientation has been adopted fully. PMID- 16271296 TI - Characterization of a new qQq-FTICR mass spectrometer for post-translational modification analysis and top-down tandem mass spectrometry of whole proteins. AB - The use of a new electrospray qQq Fourier transform ion cyclotron mass spectrometer (qQq-FTICR MS) instrument for biologic applications is described. This qQq-FTICR mass spectrometer was designed for the study of post translationally modified proteins and for top-down analysis of biologically relevant protein samples. The utility of the instrument for the analysis of phosphorylation, a common and important post-translational modification, was investigated. Phosphorylation was chosen as an example because it is ubiquitous and challenging to analyze. In addition, the use of the instrument for top-down sequencing of proteins was explored since this instrument offers particular advantages to this approach. Top-down sequencing was performed on different proteins, including commercially available proteins and biologically derived samples such as the human E2 ubiquitin conjugating enzyme, UbCH10. A good sequence tag was obtained for the human UbCH10, allowing the unambiguous identification of the protein. The instrument was built with a commercially produced front end: a focusing rf-only quadrupole (Q0), followed by a resolving quadrupole (Q1), and a LINAC quadrupole collision cell (Q2), in combination with an FTICR mass analyzer. It has utility in the analysis of samples found in substoichiometric concentrations, as ions can be isolated in the mass resolving Q1 and accumulated in Q2 before analysis in the ICR cell. The speed and efficacy of the Q2 cooling and fragmentation was demonstrated on an LCMS-compatible time scale, and detection limits for phosphopeptides in the 10 amol/muL range (pM) were demonstrated. The instrument was designed to make several fragmentation methods available, including nozzle-skimmer fragmentation, Q2 collisionally activated dissociation (Q2 CAD), multipole storage assisted dissociation (MSAD), electron capture dissociation (ECD), infrared multiphoton induced dissociation (IRMPD), and sustained off resonance irradiation (SORI) CAD, thus allowing a variety of MS(n) experiments. A particularly useful aspect of the system was the use of Q1 to isolate ions from complex mixtures with narrow windows of isolation less than 1 m/z. These features enable top-down protein analysis experiments as well structural characterization of minor components of complex mixtures. PMID- 16271299 TI - Electrophysiological and molecular genetic evidence for sympatrically occuring cryptic species in African weakly electric fishes (Teleostei: Mormyridae: Campylomormyrus). AB - For two sympatric species of African weakly electric fish, Campylomormyrus tamandua and Campylomormyrus numenius, we monitored ontogenetic differentiation in electric organ discharge (EOD) and established a molecular phylogeny, based on 2222bp from cytochrome b, the S7 ribosomal protein gene, and four flanking regions of unlinked microsatellite loci. In C. tamandua, there is one common EOD type, regardless of age and sex, whereas in C. numenius we were able to identify three different male adult EOD waveform types, which emerged from a single common EOD observed in juveniles. Two of these EOD types formed well supported clades in our phylogenetic analysis. In an independent line of evidence, we were able to affirm the classification into three groups by microsatellite data. The correct assignment and the high pairwise F(ST) values support our hypothesis that these groups are reproductively isolated. We propose that in C. numenius there are cryptic species, hidden behind similar and, at least as juveniles, identical morphs. PMID- 16271300 TI - Quantitative imaging of proteoglycan in cartilage using a gadolinium probe and microCT. AB - OBJECTIVE: Micro-computed tomography (microCT) imaging has the potential to allow the three-dimensional (3D) visualization of cartilage morphology. However, cartilage intensity on a microCT image is weak because cartilage does not strongly attenuate X-rays. This work was designed to demonstrate that exposure of cartilage to charged gadolinium compounds modifies the intensity to allow an improved visualization of cartilage morphology and the determination of proteoglycan content. DESIGN: Trypsin was used to deplete proteoglycan in bovine nasal cartilage disks. Disks were then exposed to Gd(3+), gadopentetate (Gd DTPA(2-)), or gadoteridol (Gd-HP-DO3A), and imaged with microCT. The intensities of the disks were measured from the images and compared to the actual proteoglycan content determined with a dimethylmethylene blue assay. RESULTS: Treatment of naive disks with 200 mM Gd(3+) for 24h at room temperature produced a 2.8-fold increase in intensity on microCT images. Similar treatment with 200 mM Gd-DTPA(2-) produced a 1.4-fold increase. After 2h of trypsin treatment at room temperature, the intensities of cartilage disks exposed to 20 0mM Gd(3+) decreased by 12%. Conversely, the intensities of trypsin-treated disks exposed to 200 mM Gd-DPTA(2-) increased by 15%. Trypsin treatment caused a 4% increase in the intensities of disks exposed to neutral Gd-HP-DO3A. The correlation between proteoglycan content and the microCT intensity of cartilage treated with Gd(3+) was very good (r(2)=0.81). CONCLUSIONS: Gadolinium and microCT allow an improved 3D visualization of cartilage and quantification of its proteoglycan content. PMID- 16271301 TI - Activity of Pichia pastoris alternative cis-prenyltransferase is correlated with proliferation of peroxisomes. AB - We have investigated dolichol synthesis in yeast Pichia pastoris. Growth of these cells on methanol causes peroxisome proliferation and induction of peroxisomal enzymes. Twenty-four hours methanol treatment was sufficient for the appearance of longer-chain dolichols. Less specific oleic acid induction needed 48 h for the synthesis of longer dolichol family with typical one still present. Cells cultured in non-inducing conditions for 48 h did not reveal the presence of additional dolichol family. Peroxisomes purified from oleic acid treated cells synthesize in vitro polyprenols longer by two isoprene residues than those synthesized by microsomal fraction from glucose culture. These observations lead us to suggest that chain length of dolichols synthesized in yeast cell may depend on the carbon and energy source supply which mobilizes metabolic pathways localized to different cellular compartments. PMID- 16271302 TI - Efficient gene transfer into silkworm larval tissues by a combination of sonoporation and lipofection. AB - Sonoporation (ultrasound treatment) provides a new and attractive nonviral way of in vivo gene transfer. To access the applicability of this method to the silkworm, Bombyx mori, we have compared the efficiencies of gene transfer by means of lipofection (using an appropriate agent, PDD111), sonoporation (ditto, FluoroGene), and lipofection followed by sonoporation. By these methods, a luciferase expression plasmid was found to be markedly transferred into the haemocoel of newly ecdysed fifth instar silkworm larvae, and also into other tissues although with lower rates compared with the haemocoel. In terms of luciferase activity, the efficiencies of transgene by lipofection plus sonoporation were approximately 6 (hemocytes), 20 (silk glands), 8 (mid-gut), 38 (fat body), 10 (Malpighian tubules), 33 (ovaries), and 16 (testes) times as high as those by lipofection or sonoporation alone. These results demonstrated that the present method is useful to introduce the exogenous DNA into insect organs in vivo. PMID- 16271303 TI - Effects of fluid shear stress on apoptosis of cultured human umbilical vein endothelial cells induced by LPS. AB - The objective of our research was to reveal the effects of shear stress on the apoptosis of cultured human umbilical vein endothelial cells (HUVECs) induced by lipopolysaccharide (LPS). A parallel-plate flow chamber was used to control the strength and duration of shear stress (SS), and apoptosis was measured by immunocytochemistry and radio-immunoassay. Some important conclusions were drawn. In the stationary state, apoptosis of HUVECs could be induced by LPS (50 microg/ml). An SS of 15 dyn/cm(2) could inhibit the apoptosis induced by LPS. However, an SS of 4 dyn/cm(2) had less effect on the same process. At the same time, the experiment demonstrated that the increase in IL-6 secretion by LPS can be inhibited by two different levels of shear stress. Moreover, the inhibition effect was more obvious under high level stress than under low level. We also found that the effect of shear stress on IL-8 was less effective than on IL-6. This research provides data for understanding the mechanism of the contribution of hemodynamic forces to atherosclerosis. PMID- 16271304 TI - Development of monoclonal antibodies specific to urochordate intracellular epitopes. AB - Sixteen monoclonal antibodies (MAbs) specific to 2 urochordate genera (Botryllus schlosseri and Botrylloides) intracellular epitopes were generated in mice immunized with a mixture of fresh and paraformaldehyde-fixed cells obtained from animal's blood and cells from dissociated organs. Hybridoma clones were selected by ELISA tests and immunohistochemistry assays on paraffin-embedded animal tissues. Five MAbs were tested for reactions with different zooidal organs and cell compartments; 7 MAbs were tested, separately, on 5 different botryllid colonies (3 Botryllus and 2 Botrylloides). The results revealed high polymorphism. Whereas some of the MAbs recognized, specifically, only part of the botryllid genotypes tested, others recognized only part of the cellular compartments. These MAbs will be used as an important tool in the study of botryllid ascidian immunology and developmental biology, revealing the first wide panel of MAbs specific to urochordate intracellular antigens. PMID- 16271305 TI - Stimulative effects of insulin on Toxoplasma gondii replication in 3T3-L1 cells. AB - The influence of insulin and 2-deoxy-glucose (D-glucose) on the intracellular protozoan Toxoplasma gondii replication in 3T3-L1 cells was investigated. Insulin and D-glucose had a dose-responsive mitogenic effect on intracellular T. gondii replication and development in 3T3-L1 cells. Insulin concentrations between 10( 2) and 10(-1) microg/ml combination of 4.5 g/l D-glucose in DMEM medium gave maximum stimulus to T. gondii replication. The number of tachyzoites increased rapidly, with the growth peaking typically on day 3 or 4 of culture, and then declining quickly. However, insulin, in the absence of d-glucose, had comparably less effect on T. gondii growth than two of their combination. d-glucose concentrations significantly affected the tachyzoite replication and appear to be indispensable for maintaining the host 3T3-L1 cells. PMID- 16271306 TI - Dexamethasone has pro-apoptotic effects on non-activated fresh peripheral blood mononuclear cells. AB - Apoptosis is a physiological method of cell death commonly referred to as programmed cell death. However, non-apoptotic programmed cell death, such as autophagy and programmed necrosis, has been characterized by morphological criteria. In view of the human therapeutic use of DEX, and considering that no difference in the number and/or affinity of glucocorticoid receptors in activated and non-activated lymphocytes has been reported, we decided to evaluate the effect of DEX on fresh peripheral blood mononuclear cells (PBMC). Transmission electron microscopy showed that DEX can significantly induce apoptosis in non activated PBMC. It was also observed by transmission electron microscopy that, independently of DEX treatment, PBMC also died by a process marked by extreme vacuolization and increase in cellular volume; these cells were erroneously classified as viable by flow cytometry using the 7-AAD assay. It is concluded that the DEX pro-apoptotic effect is not restricted to activated PBMC and, therefore, DEX-induced apoptosis could play either homeostatic (activated PBMC) or immunosuppressive (non-activated PBMC) roles. PMID- 16271307 TI - The nucleocytoplasmic microfilament network in protoplasts from cultured soybean cells is a plastic entity that pervades the cytoplasm except the central vacuole. AB - The microfilament network of cultured Glycine max cells (SB-1 line), and protoplasts was visualized with rhodamine-phalloidin under conditions that lysed the protoplast and changed the cell shape. The whole cell had the typical microfilament distribution of a "cage" around the nucleus, from which the large subcortical cables and transvacuolar strands radiated towards the cortex until it reached the cortical microfilament network. Upon cell wall removal, the network conserved its compartmentalization. Thus, the redistribution of the shape where the vacuole becomes a central entity, made the cytoplasm displace peripherally, but the network distribution was conserved. When protoplasts were lysed in a low osmotic medium, the vacuoles were gradually released intact. Under these conditions, the F-actin staining remained within the ghost of the cell, but none was detected in either emerging or almost completely released vacuoles. Most of the released F-actin was found in debris from the cell lysate in the form of microfilaments. When the ghosts were constrained in a coverslip with an air bubble, the shape of the ghost changed accordingly, but the microfilament network distribution remained constant. These results provide further evidence that the vacuole of plant cells does not have detectable associated F-actin. In addition, we demonstrate that the actin microfilament network is a moldable entity that can change its shape but keeps its distribution under constant conditions, in these cultured cells. PMID- 16271308 TI - eNOS genotype-dependent correlation between whole blood lead and plasma nitric oxide products concentrations. AB - Experimental data indicate that lead exposure decreases nitric oxide (NO) availability. However, no previous study has examined whether lead exposure affects plasma nitrite/nitrate (NO(x)) concentrations in humans. In addition, the T(-786)C polymorphism affects endothelial NO synthase (eNOS) expression and endogenous NO release. Here, we investigated whether there is an association between the circulating concentrations of NO(x) and the concentrations of lead in whole blood (B-Pb) and in plasma (P-Pb) from lead-exposed subjects. In addition, we also evaluated whether eNOS genotype for the T(-786)C polymorphism affects NO(x) concentrations in lead-exposed subjects. We studied 104 subjects exposed to lead who were non-smokers, 18-60 years of age, and not alcohol consumers. Genomic DNA was isolated from blood samples and genotypes for the T(-786)C polymorphism were determined by PCR and restriction fragment length digestion. Circulating NO(x) was determined by chemiluminescence. B-Pb and P-Pb were determined by inductively coupled plasma mass spectrometry and by graphite furnace atomic absorption spectrometry, respectively. No significant correlations were found between NO(x) and B-Pb and P-Pb measured in the 104 subjects (all P > 0.05). However, while no significant correlation was found in subjects with TT genotype, a negative correlation was found between plasma NO(x) and B-Pb (r = 0.230, P = 0.048) and P-Pb (r = 0.194, P = 0.110) in subjects from TC + CC genotypes group. Our study shows a negative correlation between plasma NO(x) concentrations and B Pb in carriers of the "C" allele, thus suggesting a possible mechanism possibly involved in lead exposure-induced increase in the susceptibility to cardiovascular diseases. PMID- 16271309 TI - Soluble interleukin-2 receptor as a marker for progression of coronary artery calcification in type 1 diabetes. AB - INTRODUCTION: Soluble interleukin-2 receptor (sIL2r), a marker of T cell activation, is elevated in inflammatory processes, such as rheumatoid arthritis, hepatitis and neoplasm. We explored a potential association between plasma sIL2r levels and progression of coronary artery calcification (CAC), a marker for subclinical atherosclerosis, in a prospectively followed cohort of type 1 diabetic and non-diabetic subjects, aged 20-59 years, with no history of coronary artery disease. MATERIALS AND METHODS: CAC progression was assessed by electron beam tomography over 2.6 years (range 1.6-3.2). Plasma sIL2r levels were measured in a nested case-control substudy of 98 subjects (67 diabetic, 31 non-diabetic) with and 173 subjects (84 diabetic, 89 non-diabetic) without significant CAC progression. Log-transformed sIL2r levels were analyzed by conditional logistic regression to compare subjects with and without significant CAC progression. RESULTS: SIL2r was a significant predictor for CAC progression after adjusting for presence of baseline CAC, age, gender, diabetes status, baseline calcium volume score and adiponectin (OR 1.99, 95% CI 1.09-3.61, p = 0.02 for a doubling of sIL2r level). Addition of BMI, LDL, HDL, hypertension, smoking status, HbA1c, CRP, fibrinogen, homocysteine and PAI-1 to regression models weakened but did not remove sIL2r as a predictor of CAC progression. There was no indication that this effect was different by diabetes status (p = 0.6 for diabetes-sIL2r interaction). DISCUSSION: Elevated plasma sIL2r is associated with CAC progression independent of traditional coronary artery disease risk factors in type 1 diabetic and non diabetic young adults. SIL2r should be considered as a novel marker of inflammation leading to coronary artery disease. PMID- 16271311 TI - Dale Mosbaugh. PMID- 16271310 TI - Amodiaquine resistant Plasmodium falciparum malaria in vivo is associated with selection of pfcrt 76T and pfmdr1 86Y. AB - The choice of partner drug is critical for artemisinine-based combination therapy (ACT) to remain effective and amodiaquine (AQ) is one important candidate to evaluate. We treated 81 children <5 years with uncomplicated Plasmodium falciparum malaria with AQ alone and related the treatment outcome to the possible selection of pfcrt 76T, 152T, 163S, 326S, pfmdr1 86Y and pfmrp 191H, 437S in recurrent infections (recrudescenses and re-infections) and to the blood concentration of desethylamodiaquine (DEAQ). During 21 days follow-up 28 children had a recurrent infection (9 recrudescenses, 13 re-infections and 6 mixed). Neither genotyping of the polymorphisms before treatment nor DEAQ blood concentrations could predict treatment outcome. pfcrt 76T was however significantly selected for in recurrent infections (p=0.020). pfmdr1 86Y was also selected for, but only in recrudescent infections (p=0.048). The study showed high prevalence of AQ resistant parasites in vivo, which appeared to be associated to pfcrt 76T and pfmdr1 86Y. PMID- 16271312 TI - Structural basis for mRNA Cap-Binding regulation of eukaryotic initiation factor 4E by 4E-binding protein, studied by spectroscopic, X-ray crystal structural, and molecular dynamics simulation methods. AB - Taking advantage of the Trp73 residue located close to the 4E-BP binding site of eIF4E, the interaction between the 4E-BP isoform and eIF4E was investigated by the Trp fluorescence titration method. Although no significant difference was observed among the association constants of three 4E-BP isoforms, the binding preference of 4E-BP2 over 4E-BP1 and -BP3 was shown, probably due to the effect of a 4E-BP2-specific LDRR (60-63) sequence for the binding with eIF4E. By contrast, surface plasmon resonance (SPR) analyses showed the binding preference of 4E-BP1, although the difference among the isoforms was also not significant. This inconsistency with fluorescence analysis likely resulted from the different observation points of the interaction, i.e., local and overall interactions observed by the fluorescence and SPR methods, respectively. To clarify the structural basis for these spectroscopic results, the crystal structure of the ternary complex of m7GpppA-eIF4E-4E-BP1 fragment (Thr36-Thr70) was analyzed by the X-ray diffraction method. Crystal structure analysis at 2.1 A resolution revealed that the 4E-BP1 fragment, assigned to the Pro47-Pro66 peptide moiety, adopted a reverse L-shaped conformation involving the beta sheet and alpha helical structures and was located at the root of the handle of the temple-bell shaped eIF4E through hydrophilic and hydrophobic interactions. Based on the observed binding mode, possible interactions with the three 4E-BP isoforms have been discussed. On the other hand, since the crystal structural comparison with the previously determined m7GpppA-eIF4E-4E binary complex showed that the docking of the 4E-BP1 fragment does not significantly affect the overall tertiary structure and cap-binding scaffold of eIF4E, the dynamic regulation of the cap binding of eIF4E by 4E-BP1 was investigated by molecular dynamics (MD) simulations. Consequently, the simulation suggested that (i) the helical region of the 4E-BP1 peptide is important for the binding with eIF4E, (ii) the existence of a cap structure stabilizes the binding of eIF4E with 4E-BP, (iii) the binding of 4E-BP stabilizes the cap-binding pocket of eIF4E, and (iv) the phosphorylation of Ser67 alone does not induce the separation of 4E-BP from eIF4E, but increases the structural rigidity of 4E-BP. These results provide the structural basis for the mRNA cap-binding regulation of eIF4E by 4E-BP. PMID- 16271313 TI - Psychological and cognitive effects of long-term peyote use among Native Americans. AB - BACKGROUND: Hallucinogens are widely used, both by drug abusers and by peoples of traditional cultures who ingest these substances for religious or healing purposes. However, the long-term residual psychological and cognitive effects of hallucinogens remain poorly understood. METHODS: We recruited three groups of Navajo Native Americans, age 18-45: 1) 61 Native American Church members who regularly ingested peyote, a hallucinogen-containing cactus; 2) 36 individuals with past alcohol dependence, but currently sober at least 2 months; and 3) 79 individuals reporting minimal use of peyote, alcohol, or other substances. We administered a screening interview, the Rand Mental Health Inventory (RMHI), and ten standard neuropsychological tests of memory and attentional/executive functions. RESULTS: Compared to Navajos with minimal substance use, the peyote group showed no significant deficits on the RMHI or any neuropsychological measures, whereas the former alcoholic group showed significant deficits (p < .05) on every scale of the RMHI and on two neuropsychological measures. Within the peyote group, total lifetime peyote use was not significantly associated with neuropsychological performance. CONCLUSIONS: We found no evidence of psychological or cognitive deficits among Native Americans using peyote regularly in a religious setting. It should be recognized, however, that these findings may not generalize to illicit hallucinogen users. PMID- 16271314 TI - Seasonal affective disorder and its prevention by anticipatory treatment with bupropion XL. AB - BACKGROUND: Seasonal affective disorder (SAD) can cause significant distress and impairment. No antidepressant studies have previously attempted to prevent the onset of autumn-winter depression. METHODS: Three prospective, randomized, placebo-controlled prevention trials were conducted on 1042 SAD patients, enrolled in autumn and treated while still well, across the northern US and Canada. Patients received either bupropion XL 150-300 mg or placebo daily by mouth from enrollment until spring and were then followed off medications for 8 additional weeks. Primary efficacy variables were end-of-treatment depression free rates and survival distributions of depressive recurrence. RESULTS: Despite a reported average of 13 previous seasonal depressive episodes, almost 60% of patients had never previously been treated for depression. Major depression recurrence rates during the three studies for bupropion XL and placebo groups were 19% versus 30% (p = 0.026), 13% versus 21% (p = 0.049), and 16% versus 31%; yielding a relative risk reduction across the three studies of 44% for patients taking bupropion XL. Survival analyses for depression onset also favored bupropion XL over placebo (p = .081, .057, and <.001). CONCLUSIONS: It is possible to prevent recurrence of seasonal major depressive episodes by beginning bupropion treatment early in the season while patients are still well. PMID- 16271315 TI - Medical student education in ophthalmology: crisis and opportunity. PMID- 16271316 TI - Prospective, randomized trial of trimethoprim/sulfamethoxazole vs. pyrimethamine and sulfadiazine in the treatment of ocular toxoplasmosis: discussion. PMID- 16271317 TI - Pediatric rhegmatogenous retinal detachment in East Asians. AB - PURPOSE: To describe clinical features, predisposing factors, and surgical outcomes of retinal detachment in different age groups in a pediatric population. DESIGN: Retrospective, noncomparative, interventional case series. PARTICIPANTS: A total of 296 eyes from 278 children younger than 18 years of age underwent the first surgical procedure for rhegmatogenous retinal detachment between 1983 and 2003. They were divided into 3 age groups: 38 eyes in group 1 (0-10 years), 107 eyes in group 2 (11-15 years), and 151 eyes in group 3 (16-18 years). METHODS: Predisposing factors for retinal detachment were classified into 4 categories: (1) high myopia, (2) trauma, (3) congenital or developmental structural ocular abnormalities, and (4) previous intraocular surgery. MAIN OUTCOME MEASUREMENTS: Visual acuity and retinal reattachment. RESULTS: The mean age of the patients was 14.6 years; 73.7% were boys. The mean follow-up period was 51 months. At least 1 predisposing factor could be identified in 282 (95%) of study eyes. High myopia was most common in 111 eyes (38%). Thirty-one percent of eyes experienced trauma, 17% had structural abnormalities, and 5% underwent previous intraocular surgery. Macular involvement was found in 237 eyes (80%). The initial surgery was scleral buckling alone in 224 eyes (76%). Retinal reattachment was achieved in 214 eyes (72%) after the first operation and in 250 eyes (85%) at the end of intervention. Features seen in patients with a poor surgical outcome included congenital anomaly (P<0.001), previous intraocular surgery (P = 0.007), proliferative vitreoretinopathy grade C or worse (P<0.001), macula off (P = 0.001), total retinal detachment (P<0.001), and use of silicone oil (P<0.001). CONCLUSIONS: Myopia is one of the important predisposing factors of pediatric retinal detachment in East Asians. The predisposing factors, clinical features, timing of diagnosis, and frequency of proliferative vitreoretinopathy of retinal detachment are somewhat different in the 3 age groups considered. Because of higher proportions of congenital anomaly, total retinal detachment, proliferative vitreoretinopathy, and a low success rate with poor postoperative compliance, patients in group 1 experienced the lowest final reattachment rate and the worst visual recovery. PMID- 16271318 TI - Ischemic retinopathy and uveitis in a patient with tetralogy of Fallot. AB - PURPOSE: To describe ischemic retinopathy in a patient with tetralogy of Fallot. DESIGN: Interventional case report. TESTING: Clinical and imaging evaluation. MAIN OUTCOME MEASURES: Clinical, imaging, and laboratory findings in a patient with tetralogy of Fallot. RESULTS: A 20-year-old female patient with tetralogy of Fallot had progressive visual loss of 3 weeks duration. Bilateral examination revealed dilated, tortuous, conjunctival vessels; prominent anterior chamber reaction; iris neovascularization; posterior synechia; retinal vascular tortuosity in both eyes; and inferior exudative retinal detachment. Fluorescein angiography revealed delayed retinal and choroidal filling. The working diagnosis was ischemic retinopathy with uveitis. The patient was treated for 6 months with a high-dose oral corticosteroid combined with a topical corticosteroid, a topical mydriatic, and panretinal photocoagulation. Conjunctival, vascular congestion subsided with a decrease in anterior chamber reaction. The inferior, exudative retinal detachment resolved, and vision was restored. CONCLUSIONS: Retinal ischemic syndrome combined with uveitis can develop in patients with tetralogy of Fallot. Treatment can restore vision in such patients. PMID- 16271319 TI - Primary ductal adenocarcinoma of the lacrimal gland. AB - OBJECTIVE: To report the case of a 59-year-old man with a right lacrimal gland mass, subsequently diagnosed as primary ductal adenocarcinoma of the lacrimal gland. DESIGN: Interventional case report. METHODS: We report the clinical presentation, histopathologic and immunohistochemical features, and treatment of a patient with primary ductal adenocarcinoma of the lacrimal gland. The current literature regarding this uncommon neoplasm also is reviewed. RESULTS: Excisional biopsy revealed primary ductal adenocarcinoma of the lacrimal gland. The patient subsequently underwent extensive local resection of the tumor and is awaiting radiotherapy to the orbit and neck. A literature search revealed only 3 prior reports of primary ductal adenocarcinoma of the lacrimal gland. CONCLUSIONS: Although uncommon, primary ductal adenocarcinoma of the lacrimal gland displays characteristic histologic and immunohistochemical findings that allow its distinction from the other primary lacrimal gland adenocarcinomas. Awareness and recognition of this rare malignancy may aid in further delineation of its biologic behavior, management, and prognosis. PMID- 16271320 TI - Local anesthetic in vitreoretinal surgery. PMID- 16271321 TI - Eyelid disease. PMID- 16271324 TI - Cataract surgery and Fuchs' corneal dystrophy. PMID- 16271325 TI - Submacular Surgery Trials. PMID- 16271326 TI - OCT and Band Atrophy Correlation. PMID- 16271329 TI - How can this happen? PMID- 16271330 TI - Pneumatosis cystoides intestinalis in primary intestinal pseudo-obstruction: a nonsurgical cause of pneumoperitoneum. PMID- 16271331 TI - Hepatic infarction and hepatic artery pseudoaneurysm with peritoneal bleeding after radiofrequency ablation for hepatoma. PMID- 16271332 TI - Splenic rupture after colonoscopy. PMID- 16271333 TI - Propofol versus traditional sedative agents for gastrointestinal endoscopy: a meta-analysis. AB - BACKGROUND & AIMS: Even though propofol has better recovery profile than traditional agents, its use is limited because of the perception of increased complication rates. Because an adequately powered trial comparing risk of propofol with traditional agents is lacking, we performed a meta-analysis of the current literature. METHODS: We searched Medline (1966-October 2004), EMBASE (1980-October 2004), and Cochrane controlled trials registry. The following 4 cardiopulmonary complications were assessed: hypoxia, hypotension, arrhythmias, and apnea. Procedures were divided into 3 groups: esophagogastroduodenoscopy group, colonoscopy group, and endoscopic retrograde cholangiopancreatography/endoscopic ultrasonography group. Pooled odds ratios for complications were calculated for all the procedures combined and then separately for the 3 groups. Random effects models were used for 2-proportion comparisons. RESULTS: Of the 90 citations identified, 12 original studies qualified for this meta-analysis and included 1161 patients. Of these, 634 received propofol, and 527 received midazolam, meperidine, and/or fentanyl. Most of the included studies were randomized trials of moderate quality and nonsignificant heterogeneity (Cochran Q = 4.81, P = .90). Compared with traditional sedative agents, the pooled odds ratio with the use of propofol for developing hypoxia or hypotension for all the procedures combined was 0.74 (95% confidence interval [CI], 0.44 1.24); for EGD, 0.85 (95% CI, 0.33-2.17); for colonoscopy, 0.4 (95% CI, 0.2 0.79); and for ERCP/EUS, 1.07 (95% CI, 0.38-3.01). CONCLUSIONS: Propofol sedation during colonoscopy appears to have lower odds of cardiopulmonary complications compared with traditional agents, but for other procedures, the risk of complications is similar. PMID- 16271334 TI - The genetics of irritable bowel syndrome. AB - Because of the heterogeneity in symptoms and diagnostic findings, patients with irritable bowel syndrome (IBS) remain a challenge to treat and to study. This difficulty stems from lack of understanding of the pathophysiology of this disorder. Environmental factors likely play an important role in the pathogenesis and clinical manifestations of IBS. Several recent studies suggest a genetic basis for IBS, either in etiology or predicting response to therapy. Because of interest in studying the genetic contributors to this and other functional gastrointestinal disorders, we review the literature on genetic risk factors that might explain the familial clustering of IBS. Familial aggregation studies and twin studies suggest a modest contribution of genetics to the development of IBS. Pharmacogenomic and association studies provide stronger, although far from conclusive, evidence for genetic variants that affect expression of IBS. Together, these studies suggest that a multidisciplinary approach with clinical and psychological tools, epidemiologic methods, and genetic techniques might help elucidate the molecular components leading to the common symptoms of IBS and result in better treatments for those with IBS. PMID- 16271335 TI - Short-bowel syndrome. PMID- 16271336 TI - Occult GI bleeding in NSAID users--the base of the iceberg! PMID- 16271337 TI - Time course and pattern of blood loss with ibuprofen treatment in healthy subjects. AB - BACKGROUND & AIMS: Nonselective nonsteroidal anti-inflammatory drug (NSAID) users are at increased risk of gastrointestinal bleeding. We aimed to assess the pattern and extent of fecal blood loss (FBL) with ibuprofen, which is considered to have a favorable gastrointestinal safety profile. METHODS: We conducted a post hoc analysis of 2 separate randomized, parallel-group, double-blind studies, in which ibuprofen was used as a positive control. FBL was measured by radioactive analysis of chromium-51 labeled red cells in stools during baseline and then followed by 4 weeks of treatment with ibuprofen (800 mg 3 times daily) or placebo in 68 healthy volunteers. FBL was considered significant when blood loss was >2 mL daily. RESULTS: The baseline period was identical for all subjects, with an average FBL of 0.36 mL (standard deviation, +/-0.075) per day. During the study period, all subjects receiving ibuprofen had a daily mean FBL >2 mL, with a group daily mean loss 3.64-fold greater than in the placebo group (2.55 mL [+/-3.2] vs 0.7 mL [+/-0.37], P < .001). In the ibuprofen group (n = 31), 26 subjects had between 1 and 7 random episodes of microbleeding with FBL >3 mL. Nine had a maximum FBL >10 mL (29.35 +/- 23.32 mL), and in 2 subjects blood loss reached 73 mL and 66 mL, respectively. CONCLUSIONS: Treatment with a therapeutic dose of ibuprofen, a commonly used nonselective NSAID, in healthy subjects is associated with significant FBL, which occurs randomly with spikes of bleeding, sometimes exceeding 66 mL in a single day. Chronic anemia or gastrointestinal bleeding in patients taking nonselective NSAIDs should be thoroughly investigated. PMID- 16271338 TI - Four-day Bravo pH capsule monitoring with and without proton pump inhibitor therapy. AB - BACKGROUND & AIMS: Ambulatory pH testing often is used to guide the management of reflux symptoms that do not respond to proton pump inhibitor (PPI) therapy and to evaluate the contribution of acid reflux to atypical symptoms. Controversy exists as to whether such clinical studies are performed optimally off or on PPI therapy. The aim of the present study was to determine the feasibility of 4-day pH recordings using a pH system that would encompass time periods both before and during PPI therapy. METHODS: Eighteen patients underwent 4-day ambulatory pH testing using 2 separate receivers calibrated to a single Bravo pH capsule. Rabeprazole was administered on days 2-4 of the study (20 mg orally twice a day). RESULTS: Indications for pH testing were refractory heartburn, chest pain, or chronic cough. pH recordings showed that 9 patients (53%) had esophageal acid exposure values that exceeded 4% on day 1 and 7 patients (41%) had values that exceeded 5.3%. Patients showed significant and progressive reductions in acid exposure on days 2-4 of the recording period. Of the 7 patients with quantitatively abnormal levels of acid exposure on day 1, 86% had normalization by day 3. CONCLUSIONS: Prolonged, esophageal pH recordings using the Bravo pH system are feasible and allow for combined testing both off and on a therapeutic trial of PPI. Such studies may allow for the acquisition of complementary information in a single test that may be useful in the management of patients with suspected gastroesophageal reflux disease symptoms. PMID- 16271339 TI - Barrett's esophagus in women: demographic features and progression to high-grade dysplasia and cancer. AB - BACKGROUND & AIMS: Barrett's esophagus is traditionally considered a disease of older white men. The aims of this study were to compare the demographic features of Barrett's esophagus in men and women and to determine the prevalence and incidence of high-grade dysplasia and cancer in these patients. METHODS: All patients enrolled in the Cleveland Clinic Barrett's Esophagus Registry from 1979 2002 were studied. Age, ethnicity, number of endoscopies, hiatal hernia size, length of Barrett's segment, and prevalence and incidence of high-grade dysplasia and cancer were compared between men and women. RESULTS: There were 839 patients in the registry (628 men and 211 women). Barrett's segment length was greater in men than in women (mean, 5.06 +/- 4.2 vs 4.05 +/- 3.27 cm, respectively; P = .003). There were no significant differences for other parameters. There were 114 prevalence cases of high-grade dysplasia or cancer (96 men, 18 women). Women were less likely to have prevalent high-grade dysplasia or cancer than men (odds ratio, 0.52; 95% confidence interval, 0.31-0.88; P = .015). There were 13 incidence cases of high-grade dysplasia or cancer (11 men, 2 women) during a mean follow-up of 4.72 years, which was similar in both genders with an incidence rate of 1 in 179 patient-years of follow-up for women and 1 in 91 patient-years of follow-up in men. CONCLUSIONS: Twenty-five percent of patients in our registry are women. The length of Barrett's esophagus is greater in men than in women, but other features are similar. The prevalence of high-grade dysplasia/cancer in women is approximately half that of men. Incidence rates for high-grade dysplasia/cancer are similar in men and women, although the number of cases is small. PMID- 16271340 TI - Contribution of ascites to impaired gastric function and nutritional intake in patients with cirrhosis and ascites. AB - BACKGROUND & AIMS: Protein calorie malnutrition and weight loss are common among patients with cirrhosis and ascites. The cause of these symptoms is unclear, with several putative mechanisms proposed. The primary aims of this study were to compare gastric volumes and accommodation between patients with cirrhosis complicated by ascites and healthy controls, and to evaluate the effect of large volume paracentesis in the patient group. METHODS: Patients with cirrhosis and ascites underwent assessment of gastric volumes as measured by single-photon emission computed tomography, gastric sensation assessed by a validated nutrient drink test, and a 3-day assessment of caloric intake before and after large volume paracentesis. Age- and sex-adjusted linear regression models were used to compare gastric volumes and accommodation ratios between patients and healthy volunteers. Paired Wilcoxon rank-sum tests were used to compare gastric measures before and after paracentesis among the patient group. RESULTS: Fifteen patients (median age, 54 y) were compared with 112 healthy (age- and sex-matched) controls. Median postprandial gastric volumes (627 mL patients vs 721 healthy controls) and gastric accommodation were reduced significantly in patients compared with healthy controls (P = .02 and .006, respectively). After paracentesis: (1) fasting gastric volumes were increased (median 312 mL post- vs 241 mL pre-, P = .04), (2) patients tolerated ingestion of larger maximum volumes (median 964 mL post- vs 738 mL pre-, P = .04), and (3) caloric intake was increased (median 34% kcal post- vs 3110 kcal pre-, P = .005). CONCLUSIONS: Postprandial gastric volumes and accommodation ratios are reduced in patients with cirrhosis and ascites compared with healthy controls. In addition, large volume paracentesis increases fasting gastric volumes, volumes ingested until maximal satiation, and caloric intake. PMID- 16271341 TI - Bowel frequency and defecatory patterns in children: a prospective nationwide survey. AB - BACKGROUND & AIMS: Very little is known about several aspects of bowel habits in the general pediatric population and the aim of this nationwide survey was to assess bowel frequency and modalities of defecation in children. METHODS: The survey was conducted by 58 pediatricians who were selected randomly and distributed evenly throughout Italy. The following items were reported by each pediatrician in a standardized questionnaire: sociodemographic data, frequency and modalities of bowel evacuation, and anorectal disorders. RESULTS: The response rate of available completed questionnaires was distributed evenly for the 1-year age group and was 94% (number of patients, 2680; 1-2 years, 442; females, 49.8%). Mean bowel frequency did not vary in the first 2 years of life, it decreased (P = .00001) after the second year, and remained stable until the 12th year; it did not differ between sexes. Mean bowel frequency was reduced significantly in children, both in those younger or older than 2 years, with a positive history of constipation in the parents (P = .00002). Bowel frequency was inversely correlated with the number of persons living and the number of rooms in the child's house (P < .05, P = .008, respectively). Stool consistency, duration of evacuation, and frequency of episodes of painful defecation showed an inverse relationship (P < .001) with bowel frequency. Bowel frequency was significantly lower (P < .001) in children with anorectal disorders. CONCLUSIONS: In Italian children, bowel frequency does not differ between sexes but it differed between the first 2 years of life and age older than 2. Anorectal disorders increase as bowel frequency decreases. PMID- 16271342 TI - A randomized trial of nicotine enemas for active ulcerative colitis. AB - BACKGROUND & AIMS: Ulcerative colitis (UC) is largely a disease of nonsmokers in which transdermal nicotine improves the symptoms but often causes adverse events (AEs). Nicotine enemas cause fewer AEs and were used as supplemental treatment for active UC. METHODS: We treated 104 patients with active UC with either 6-mg nicotine enemas or placebo enemas for 6 weeks in a randomized double-blind study. Patients continued their oral therapy, if any, for UC: 68 patients were taking mesalamine, 15 patients were taking prednisolone, and 12 patients were taking thiopurines during the study. Clinical, sigmoidoscopic, and histologic assessments were made at baseline and at the end of the study and symptoms were recorded daily on a diary card. The primary end point was induction of clinical remission and clinical improvement also was measured by the UC disease activity index. After the study, patients then used nicotine enemas daily for 4 weeks and sigmoidoscopy with a biopsy examination was repeated. AEs and salivary cotinine levels were monitored throughout the study. RESULTS: Clinical remission was achieved in 14 of 52 (27%) patients on active treatment and 14 of 43 (33%) patients on placebo (P = .55). The UC disease activity index improved by 1.45 points in the active group and by 1.65 points for those on placebo (P = .88). Only 1 patient discontinued treatment because of an AE (abdominal pain). In the 47 patients taking mesalamine only, active treatment conferred benefit that was not statistically significant; disease remission occurred in 9 of 25 patients on active therapy and 4 of 21 patients on placebo (P = .20). CONCLUSIONS: Six milligram nicotine enemas were well tolerated but were not found to be efficacious for active UC. PMID- 16271343 TI - Refining molecular analysis in the pathways of colorectal carcinogenesis. AB - BACKGROUND & AIMS: In the stepwise model, specific genetic and epigenetic changes accumulate as colorectal adenomas progress to carcinomas (CRCs). CRCs also acquire global phenotypes, particularly microsatellite instability (MSI) and aneuploidy/polyploidy (chromosomal instability, CIN). Few changes specific to MSI low or CIN+ cancers have been established. METHODS: We investigated 100 CRCs for: mutations and loss of heterozygosity (LOH) where appropriate, of APC, K-ras, BRAF, SMAD4, and p53; deletion on 5q around APC and 18q around SMAD4; total chromosomal-scale losses and gains; MSI; and CIN. RESULTS: As expected, CIN- cancers had fewer chromosomal changes overall than CIN+ lesions, but after correcting for this, 5q deletions alone predicted CIN+ status. 5q deletions were not, however, significantly associated with APC mutations, which were equally frequent in CIN+ and CIN- tumors. We therefore found no evidence to show that mutant APC promotes CIN. p53 mutations/LOH were more common in CIN+ than CIN- lesions, and all chromosomal amplifications were in CIN+ tumors. CIN- cancers could be subdivided according to the total number of chromosomal-scale changes into CIN-low and CIN-stable groups; 18q deletion was the best predictor, being present in nearly all CIN-low lesions and almost no CIN-stable tumors. MSI-low was not associated with CIN, any specific mutation, a mutational signature, or clinicopathologic characteristic. CONCLUSIONS: Overall, the components of the stepwise model (APC, K-ras, and p53 mutations, plus 18q LOH) tended to co-occur randomly. We propose an updated version of this model comprising 4 pathways of CRC pathogenesis, on the basis of 5q/18q deletions, MSI (high/low), and CIN (high/low/stable). PMID- 16271344 TI - The effect of prior colonic imaging on endoscopic productivity: potential impact of computed tomographic colonography. AB - BACKGROUND & AIMS: If computed tomographic colonography (CTC) is used for primary colorectal cancer (CRC) screening with a small polyp size threshold to define a CTC study as positive, a substantial portion of all colonoscopies performed annually will be to follow up positive CTC examinations. Moreover, the majority of positive CTC examinations would be false positives (FP). This case-control study was undertaken to test the hypothesis that colonoscopy examinations resulting from FP CTC studies would take longer to complete than negative screening colonoscopies. METHODS: Endoscopic records of a large, urban hospital were reviewed to identify all patients who had either a positive barium enema (BE) study or flexible sigmoidoscopy (FS) and a negative follow-up colonoscopy examination (these patients were used as surrogates for CTC FP cases). For each of the 28 FP patients or cases identified, 2 screening colonoscopies performed by the same endoscopist within the same time period were identified and used as matched controls. A two-way analysis of variance test was performed to assess for differences in time to complete colonoscopies between these 2 groups, controlling for the individual endoscopist. RESULTS: FP colonoscopies took an average of 24.0 minutes to complete, whereas negative screening colonoscopies took 14.9 minutes; FP colonoscopies required 61% more active time to complete. This highly statistically significant difference (P < .0001) persisted with subset analyses that only included BE or FS cases and when fellow or surgeon cases were excluded. CONCLUSIONS: FP colonoscopies take longer to perform than negative screening colonoscopies. If CTC is implemented as the primary modality for CRC screening, these FP examinations could comprise a substantial percentage of the colonoscopies performed, potentially leading to a significant decrease in endoscopic productivity. PMID- 16271345 TI - Long-term results of extracorporeal shockwave lithotripsy and endoscopic therapy for pancreatic stones. AB - BACKGROUND & AIMS: The aim of this study was to evaluate the short- and long-term results of extracorporeal shockwave lithotripsy (ESWL) and endoscopic therapy for pancreatic stones. METHODS: A total of 117 patients with pancreatic stones underwent ESWL and endoscopic treatment in our institute. Seventy patients who were followed-up for over 3 years after treatment were evaluated retrospectively. RESULTS: Immediate pain relief was achieved in 97% and complete removal of stones was achieved in 56%. During the long-term follow-up evaluation, 49 of 70 patients continued to be asymptomatic. Pain recurred more frequently in patients with incomplete removal than in those with complete removal (P < .05). Twenty-one patients who became symptomatic during the follow-up period underwent additional therapy, and pain relief was attained without surgery in all of them. Both endocrine and exocrine function deteriorated after the long-term follow-up period (P < .05). CONCLUSIONS: ESWL and endoscopic treatment of pancreatic stones proved to be effective for long-term pain relief, especially in patients in whom stones were removed completely at initial therapy. PMID- 16271346 TI - Long-term follow-up of intraductal papillary mucinous neoplasm of the pancreas with ultrasonography. AB - BACKGROUND & AIMS: Most intraductal papillary mucinous neoplasms of the pancreas (IPMNs) have a favorable prognosis. This study was undertaken to clarify the clinical course of IPMNs and to set the criteria for surgical treatment on the basis of long-term follow-up by periodic transabdominal ultrasonography (US). METHODS: Eighty-one patients with IPMN were periodically subjected to US (>3 years); 27 were reviewed retrospectively (12 with benign neoplasms [adenoma, borderline] and 15 with malignant tumors [carcinoma in situ, invasive cancer]) and 54 prospectively. US examination was focused on the main pancreatic duct (MPD) diameter, cyst diameter, and presence or absence and height of the protruding lesion. Differences between the benign and malignant groups were examined to set the criteria for surgical treatment. Follow-up results of prospective patients were investigated. RESULTS: The increase of the MPD or the cyst diameter was significantly greater in the malignant group (P < .01). Maximum increases of the MPD diameter by 2.2 mm/year, the cyst diameter by 11.3 mm/year, and emergence or increase of the height of the protruding lesion by 3.3 mm/year were predominantly observed in the malignant group (accuracy, 90.9%, 81.3%, and 81.5%, respectively). The majority of the prospective patients showed no significant changes of these parameters; however, 2 patients (3.7%) were operated on, with the post-surgery histopathologic diagnosis of cancer. CONCLUSIONS: Periodic imaging follow-up is useful to detect a malignant IPMN. Changes in MPD diameter, cyst diameter, and/or size of the protruding lesion are the practical criteria for selecting surgery. PMID- 16271347 TI - Endoscopic therapy of posttransplant biliary stenoses after right-sided adult living donor liver transplantation. AB - BACKGROUND & AIMS: Endoscopic treatment of biliary strictures after liver transplantation is a therapeutic challenge. In particular, outcomes of endoscopic therapy of biliary complications in the case of duct-to-duct anastomosis after living related liver transplantation are limited. The aim of this study was to evaluate the feasibility and success of an endoscopic treatment approach to posttransplant biliary strictures (PTBS) after right-sided living donor liver transplantation (RLDLT) with duct-to-duct anastomosis. METHODS: Ninety patients who received adult-to-adult RLDLT in our center were screened retrospectively with respect to endoscopic treatment of PTBS. Therapy was judged as successful when cholestasis parameters returned to normal and bile duct narrowing was reduced significantly after the completion of therapy. RESULTS: Forty of 90 RLDLT patients received duct-to-duct anastomosis, 12 (30%) showed PTBS. Seven of 12 patients were treated successfully by endoscopy; the remaining 5 patients were treated primarily by surgery. Most patients were treated by balloon dilatation followed by insertion of endoprostheses. A median of 2.5 dilatation sessions were necessary and the median treatment duration was 8 months. One patient developed endoscopy-treatable recurrent stenosis, no surgical intervention was necessary. Mild pancreatitis occurred in 7.9% and cholangitis in 5.3% of the procedures. One minor bleeding episode occurred during sphincterotomy. Bleeding was managed endoscopically. CONCLUSIONS: Endoscopic therapy of adult-to-adult right living related liver transplantation with duct-to-duct anastomosis is feasible and frequently is successful. The duct-to-duct anastomosis offers the possibility of endoscopic treatment. Endoscopic treatment of posttransplant biliary strictures is safe, with a low specific complication rate. PMID- 16271348 TI - Influence of alcohol on the progression of hepatitis C virus infection: a meta analysis. AB - BACKGROUND & AIMS: A convincing, yet inconsistent, pattern has emerged that demonstrates increased progression of HCV-related liver disease with heavy alcohol use. The aim was to perform a meta-analysis to quantify the effect of alcohol on cirrhosis risk among persons infected with HCV. METHODS: A meta analysis of 20 articles, involving more than 15,000 HCV chronically infected persons, published between 1995 and 2004 was undertaken to explore the relationship between advanced liver disease and the consumption of alcohol. RESULTS: The pooled relative risk of cirrhosis associated with heavy alcohol intake (defined in the range of at least 210-560 g per week) was 2.33 (95% confidence interval, 1.67-3.26) by the random effects model. The risk of HCV related liver disease associated with heavy alcohol intake increased with severity of the outcome; the lowest (1.63; 95% confidence interval, 1.22-2.17) and highest (3.54; 2.14-5.85) pooled relative risk estimates were obtained for advanced fibrosis and decompensated cirrhosis, respectively. The regression effect of alcohol might, however, be underestimated in studies investigating the risk of HCV-related cirrhosis because they necessarily include patients undergoing liver biopsy and could therefore under-represent heavy alcohol users. CONCLUSIONS: The evidence overwhelmingly shows a worsened outcome for those with chronic HCV and concurrent alcohol use. Studies varied widely in their definition of significant alcohol intake, and so the true threshold above which alcohol accelerates HCV disease progression remains uncertain. Alcohol consumption should be minimized as much as possible in those who have chronic HCV until a safe threshold is more definitively determined. PMID- 16271349 TI - Involvement of gap junctions in placental functions and development. AB - Connexin (Cx) expression and gap junctional intercellular communication (GJIC) are involved in development and differentiation processes. Mediating exchanges between mother and fetus, the placenta is formed when fetal membranes are apposed or even fusing or destroying the uterine mucosa. Therefore, an extraordinary variability of placental structures is observed throughout the mammalian species. This variability affect mainly, the maternofetal blood flow interrelationships, the kind and number of tissue layers separating maternal and fetal bloods, the trophoblast invasiveness and the formation of a syncytium (syncytiotrophoblast). Here, the expression, the localisation and the possible role of Cx and GJIC in placental functions and development are discussed. In rodents, gene knock out in mice have vastly improved our understanding of the role of Cx genes in mouse placental development: Cx26 in transplacental uptake of glucose, Cx31 in the proliferative process of trophoblastic cells and Cx45 in placental vascularisation. In human, it appears that Cx43 allows a GJIC required for the fusion process of cytotrophoblastic cells leading to the formation of the syncytiotrophoblast, the site of the numerous placental functions. On other hands, Cx40 plays a critical role in the switch from a proliferative to an invasive phenotype of the trophoblastic cells invading the endometrium. Owing to the striking diversity of Cx expression in placental structures, we must be careful when extrapolating findings from one species to another. PMID- 16271350 TI - Combined cold, acid, ethanol shocks in Oenococcus oeni: effects on membrane fluidity and cell viability. AB - The effects of combined cold, acid and ethanol on the membrane physical state and on the survival of Oenococcus oeni were investigated. Membrane fluidity was monitored on intact whole O. oeni cells subjected to single and combined cold, acid and ethanol shocks by using fluorescence anisotropy with 1,6-diphenyl-1,3,5 hexatriene (DPH) as a probe. Results showed that cold shocks (14 and 8 degrees C) strongly rigidified plasma membrane but did not affect cell survival. In contrast, ethanol shocks (10-14% v/v) induced instantaneous membrane fluidisation followed by rigidification and resulted in low viability. Acid shocks (pH 4.0 and pH 3.0) exerted a rigidifying effect on membrane without affecting cell viability. Whatever the shock orders, combined cold (14 degrees C) and ethanol (14% v/v) shocks resulted in strong membrane rigidification. Interestingly, O. oeni survived combined cold and ethanol shocks more efficiently than single ethanol shock. Membrane rigidification was induced by ethanol-and-acid (10% v/v - pH 3.5) shock and correlated with total cell death. In contrast, O. oeni recovered its viability when subjected to cold (8 degrees C)-then-ethanol-and acid shock which strongly rigidified the membrane. Our results suggested a positive short-term effect of combined cold, acid and ethanol shocks on membrane fluidity and viability of O. oeni. PMID- 16271351 TI - Update on the domain architectures of NLRs and R proteins. PMID- 16271352 TI - Isoflavone genistein and daidzein up-regulate LPS-induced inducible nitric oxide synthase activity through estrogen receptor pathway in RAW264.7 cells. AB - Isoflavones, such as genistein and daidzein, are found in abundance in soybeans. These plant-derived substances have estrogenic activities and can bind to the estrogen receptors (ERs). In this study, we investigated that the effects of 17beta-estradiol (E2), genistein and daidzein on nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) activity in RAW264.7 cells. We found that these isoflavones significantly increased lipopolysaccharide-induced NO production and iNOS expression as much as E2 at physiological concentrations. Moreover, E2 and isoflavone enhanced the production of tumor necrosis factor alpha that is one of the important cytokines regarding NO production. The enhancing effects of E2 and isoflavones on NO production were markedly inhibited by not only N(G)-nitro-L-arginine methyl ester (an inhibitor of NOS), but also ICI 182780 (ERs antagonist). Two types of ERs were identified as ERalpha and ERbeta. An ERalpha agonist could increase iNOS expression in RAW264.7 cells, while an ERbeta agonist could not. In conclusion, our results suggest E2, genistein and daidzein activate iNOS, and then up-regulate NO production. This enhancing effect is aroused through ERalpha pathway in RAW264.7 cells. PMID- 16271353 TI - Transport deficient (TR-) hyperbilirubinemic rats are resistant to acetaminophen hepatotoxicity. AB - The biliary excretion of acetaminophen (APAP) is reduced in transport deficient (TR-) hyperbilirubinemic rats lacking the multidrug resistance-associated protein 2 (Mrp2). This mutant strain of Wistar rats has impaired biliary excretion of organic anions and increased hepatic glutathione. The rational for this study was to determine if there is an altered risk for liver damage by APAP in the absence of Mrp2. Therefore, the susceptibility of TR- rats to APAP hepatotoxicity was investigated. Male Wistar and TR- rats were fasted overnight before APAP treatment (1 g/kg). Hepatotoxicity was assessed 24 h later by plasma sorbitol dehydrogenase activity and histopathology. In other studies, TR- rats received buthionine sulfoximine before APAP to reduce hepatic glutathione to values similar to those in Wistar rats. mRNA expression of APAP metabolizing enzymes was also measured in naive animals. Wistar rats treated with APAP showed significant elevations in plasma sorbitol dehydrogenase activity, while no increases in enzyme activity were observed in TR- rats. Histopathology was in agreement. Hepatic non-protein sulfhydryls were significantly lower in Wistar rats receiving APAP than in TR- rats. TR- rats treated with buthionine sulfoximine and APAP showed dramatic increases in hepatotoxicity. TR- rats had increased mRNA expression of several APAP metabolizing enzymes. Mrp2 expression not only is important in biliary excretion, but also influences the toxic potential of reactive intermediates by controlling intrahepatic GSH and possibly drug metabolism. PMID- 16271354 TI - Effects of early weaning and social isolation on the expression of glucocorticoid and mineralocorticoid receptor and 11beta-hydroxysteroid dehydrogenase 1 and 2 mRNAs in the frontal cortex and hippocampus of piglets. AB - Pigs weaned at young ages show more abnormal and aggressive behaviors and cognitive deficits compared to later weaned pigs. We investigated the effects of age, weaning and/or social isolation on the expression of genes regulating glucocorticoid response [glucocorticoid receptor (GR), mineralocorticoid receptor (MR), 11beta-hydroxysteroid dehydrogenases 1 and 2 (11beta-HSD1 and 11beta-HSD2)] in the frontal cortex and hippocampus. Early- (EW; n = 6) and conventionally weaned (CW; n = 6) piglets were weaned at 10 and 21 days after birth, respectively. Non-weaned (NW) piglets of both ages (NW; n = 6/group) remained with their dams. Immediately before euthanasia, half of CW, EW and NW animals were socially isolated for 15 min at 12 (EW, NW) and 23 (CW, NW) days of age. Differences in amounts of 11beta-HSD1, 11beta-HSD2, GR and MR mRNA were determined by quantitative real-time RT-PCR and data subjected to multivariate linear mixed model analysis. When compared with NW piglets at 12 days of age, the hippocampi of EW piglets showed decreased gene expression (P < 0.01). Social isolation decreased gene expression (P < 0.05) in the frontal cortex of all piglets. Twelve-day-old piglets showed higher MR mRNA in the frontal cortex (P < 0.01) and lower 11beta-HSD2 and GR mRNA (P < 0.05) in the hippocampus compared to 23-day-old animals. Results indicate that EW affected the hippocampus of piglets at 12 days of age, while social isolation affected frontal cortex regardless of age. These results may be correlated with behavioral and cognitive changes reported in EW piglets. PMID- 16271355 TI - Characterization of hippocampal theta rhythm in wild-type mice and glutamate receptor subunit delta2 mutant mice during eyeblink conditioning with a short trace interval. AB - We have shown that glutamate receptor subunit delta2 (GluRdelta2) null mutant mice, which have serious morphological and functional deficiencies in the cerebellar cortex, are severely impaired in delay eyeblink conditioning but not in trace eyeblink conditioning, even with a 0-trace interval. Such 0-trace conditioning does not depend critically on the hippocampus in wild-type mice, but it does in GluRdelta2 mutant mice. Here we examined the hippocampal electroencephalogram (EEG) during 0-trace conditioning in GluRdelta2 mutant and wild-type mice. During the apparatus habituation sessions, the total hippocampal theta activity (4-12 Hz) of GluRdelta2 mutant mice was less than that of wild type mice. Activity in the higher frequency band (8-12 Hz, type 1) in GluRdelta2 mutant mice was significantly less than it was in wild-type mice, but activity in the lower frequency band (4-8 Hz, type 2) was not. As learning proceeded during the acquisition sessions, the total theta activity decreased in many of the wild type mice, while this phenomenon was less prominent in GluRdelta2 mutant mice. Further analysis showed that the type 1 activity in wild-type mice increased in the early sessions and then decreased, while that in GluRdelta2 mutant mice did not change. Type 2 activity tended to decrease in both types of mice as the conditioning proceeded. These results indicate that the distribution of hippocampal EEG frequency and its properties during conditioning are different between wild-type and GluRdelta2 mutant mice, suggesting that the cerebellar cortical dysfunction may cause an alteration in the electrophysiological characteristics of the hippocampus. PMID- 16271356 TI - Phorbol ester induced short- and long-term permeabilization of the blood-CSF barrier in vitro. AB - Interconnected by tight junctions, the epithelial cells of the choroid plexus form a barrier separating the cerebrospinal fluid (CSF) from blood. Using an in vitro model based on porcine choroid plexus epithelial cells (PCPEC), we investigated the influence of PKC activating phorbol 12-myristate 13-acetate (PMA) on barrier properties and analyzed mechanisms involved in the regulation of barrier tightness. Applied in concentrations of 5-25 nM, PMA induced a fast and lasting decrease of the transepithelial electrical resistance (TER), which could be blocked by rottlerin, indicating the involvement of PKCdelta in signal transduction. The immediate impairment of barrier integrity was accompanied by dephosphorylation of occludin and formation of actin bundles. Moreover, in the presence of at least 25 nM PMA, changes of cell shape as well as discontinuities of tight junction strands were observed, suggesting the disruption of cell-cell contacts. Exposure to PMA for 1-2 days additionally induced down-regulation of claudin-2 and up-regulation of barrier modulating matrix metalloproteinase (MMP) 9, respectively. The results show that different interconnected mechanisms directly and indirectly targeting at the tight junctions are released by PMA contributing to the short-term and long-term decrease of TER and opening of the blood-CSF barrier in vitro. PMID- 16271357 TI - Erybraedin C and bitucarpin A, two structurally related pterocarpans purified from Bituminaria bituminosa, induced apoptosis in human colon adenocarcinoma cell lines MMR- and p53-proficient and -deficient in a dose-, time-, and structure dependent fashion. AB - Pterocarpans, the second group of natural isoflavonoids, have received considerable interest on account of their medicinal properties. These drugs are employed as antitoxins, but display antifungal, antiviral and antibacterial properties as well. Erybraedin C and bitucarpin A are two new structurally related pterocarpans recently purified and characterized. Bitucarpin A differs from erybraedin C for the absence of a prenyl group in 5' position and the presence of a methoxylate hydroxyl group in 7, 4' positions. These compounds proved not to be clastogens in human lymphocytes per se but displayed anticlastogenic activity against mytomicin C and bleomycin C. Here we extended the study of their antiproliferative and apoptosis-inducing mechanism on human cell lines. Two human adenocarcinoma cell lines, LoVo and HT29, as examples of slow-growing solid tumors, proficient and deficient in mismatch repair system (MMR), p53 and Bcl-2, were used to evaluate the cytotoxicity of the drugs and their effects on the cell cycle, measured by flow cytometry. Erybraedin C similarly affects the survival of HT29 (MMR +/+, p53 -/- and Bcl-2 +/+) and LoVo (MMR -/-, p53 +/+ and Bcl-2 -/-) cells (LD(50): 1.94 and 1.73 microg/ml, respectively). By contrast, bitucarpin A exhibits a differential cytotoxicity in the cell lines (LD(50): 6.00 microg/ml, HT29, and 1.84 microg/ml, LoVo). The cell cycle distributions of the LoVo and HT29 cells treated with erybraedin C lacked a specific checkpoint arrest, whereas they underwent a characteristic sub-G(1) peak, time- and drug-concentration dependent. So that apoptotic process induced by erybraedin C in both adenocarcinoma cell lines is independent of cell cycle arrest and of phenotypic status of the cells as well. By contrast, bitucarpin A affects cell cycle progression on both cell lines, inducing a transient block in G(0)/G(1) along 24-96 h, and induces apoptosis with a cell density and treatment time dependency. Similar results were obtained with the positive control drug etoposide. The programmed cellular death on human adenocarcinoma cell lines may be efficiently activated, via a topoisomerase II poison pattern, by erybraedin C, the drug containing regio-specific hydroxyl and prenyl groups. The apoptotic effect induced by the methoxylated bitucarpin A proved to be conditioned by cell density and required higher dose (5-fold-LD(50)) and longer treatment time. The present study provides evidences that erybraedin C may act as a potent growth inhibitory compound, at low and high cell density, comparable to other clinically important antineoplastic natural drugs including etoposide, on human colon adenocarcinoma cells. Bitucarpin A proved less active because it was conditioned by cell density effect, but this finding may represent a clinical advantage against early micrometastatic diseases. PMID- 16271358 TI - Survival curves for microbial species stored by freeze-drying. AB - The survival of a variety of species of microorganism following storage for up to 20 years has been analyzed. The organisms were freeze-dried, sealed in ampoules under vacuum (<1 Pa) and stored in the dark at 5 degrees C. The yeast that was tested, Saccharomyces cerevisiae, showed only 8% survival when recovered shortly after freeze-drying, but subsequent loss during storage was the least among all the tested microorganisms. The decrease in the logarithm of survival per year (log survival) was -0.010, which corresponds to a survival rate of 97.7% per year. The Gram-negative bacteria tested, Escherichia coli, Pseudomonas putida, and Enterobacter cloacae, showed 42.6, 33.5, and 50.8% survival shortly after freeze-drying, which was higher than the corresponding survival of S. cerevisiae, but the subsequent loss during storage was greater than S. cerevisiae, the log survival figures being -0.041, -0.058, and -0.073 per year. These values correspond to survival rates of 91.0, 87.5, and 84.5% each year. The Gram positive bacteria tested, Lactobacillus acidophilus and Enteroccoccus faecium, showed 62.5 and 85.2% survival shortly after freeze-drying, which was even higher than that of the Gram-negative species, and these organisms also showed better survival during storage than Gram-negative bacteria; their log survival rates were -0.018 and -0.016 per year, which corresponded to survival rates of almost 96% per year. Comparison of these results with other published data for different drying conditions suggests that survival during storage is strongly influenced by the degree of vacuum under which the ampoules were sealed. The excellent survival after freeze-drying of each species might be attributable to the high level of desiccation and to sealing under vacuum. PMID- 16271359 TI - Modulation of ATP-mediated contractions of the rat vas deferens through presynaptic cannabinoid receptors. AB - The effect of R-(+)-[2,3-dihydro-5-methyl-3-[(morpholiny)methyl]pyrolol[1,2,3-de] 1,4-benzoxazin-yl]-(1-naphthalenyl)methanone mesylate (WIN 55,212-2; a cannabinoid receptor agonist) was investigated on contractions of the bisected (epididymal and prostatic portions) rat vas deferens to assess the role of cannabinoid receptors in sympathetic ATP neurotransmission. WIN 55,212-2 inhibited the electrically induced contractions in both portions of the rat vas deferens. In the presence of the alpha1-adrenoreceptor antagonist prazosin, electrical stimulation produces a contraction mediated exclusively by ATP. In this condition, WIN 55,212-2 in the prostatic portion elicited a concentration dependent inhibition that was antagonized by N-piperidinyl-[8-chloro-1-(2,4 dichlorophenyl)-1,4,5,6-tetrahydrobenzo[6,7]cyclohepta[1,2-c]pyrazole-3 carboxamide] (NESS 0327), a selective cannabinoid CB1 receptor antagonist. NESS 0327 caused a parallel dextral displacement of the WIN 55,212-2 concentration response curve. It is suggested that activation of pre-junctional cannabinoid receptors on sympathetic nerves of the vas deferens modulates ATP neurotransmission. PMID- 16271360 TI - Drug-eluting stents: sirolimus and paclitaxel differentially affect cultured cells and injured arteries. AB - Sirolimus and paclitaxel eluted from stents inhibit cell proliferation and other cellular processes by dramatically different mechanisms. In this study, the effects of sirolimus and paclitaxel on cultured human coronary artery smooth muscle and endothelial cell function or cell cycle changes in balloon-injured arteries were directly compared. Both sirolimus and paclitaxel inhibited smooth muscle and endothelial cell proliferation. However, only paclitaxel inhibited smooth muscle and endothelial cell migration at low (nM) concentrations. Sirolimus arrested smooth muscle and endothelial cells in the G0/G1 phase of the cell cycle without inducing apoptosis while paclitaxel produced apoptosis in both cell types at low nanomolar concentrations. Although both agents blocked neointimal formation, sirolimus applied locally to injured rat carotid arteries increased the percentage of cycling vascular cells in G0/G1 without inducing apoptosis while paclitaxel increased the percentage of cycling cells in S and G2/M phases while inducing apoptosis. These results suggest that sirolimus reduces neointimal hyperplasia through a cytostatic mechanism while paclitaxel produces apoptotic cell death. PMID- 16271361 TI - Flow cytometry: a new approach for the molecular profiling of breast cancer. AB - The established method in prognosis of breast cancer includes detection of molecular markers, such as the estrogen receptor (ER), progesterone receptor (PR), and HER-2/neu. These markers are routinely checked via immunohistochemistry (IHC). HER-2/neu is also detected by fluorescent in situ hybridization (FISH). Flow cytometric analysis has not yet been used for detection of such markers. Flow cytometry was performed on four established breast cancer cell lines: MCF7, T47D, BT474, MDA-MB-231, and on one normal breast epithelial cell line: MCF10A. Flow cytometric analysis was used for the detection of ER, PR, HER-2/neu, epidermal growth factor receptor (EGFR), and E-cadherin. Currently, EGFR and E cadherin are not standard predictive factors in determining survival of breast cancer patients, but both may be beneficial to prognosis. Cells undergoing flow cytometric analysis lost marker expression with increasing passage number. The highest expression was found at cells passaged 0-1 times. MCF7, T47D, and BT474 all had similar marker expression patterns. E-cadherin demonstrated a strongly positive pattern with marker expression of 85-92% among the three cell lines. ER, PR, and HER-2/neu demonstrated a weakly positive expression pattern when compared with E-cadherin. Marker expression ranged from 15 to 61%. These three cell lines were almost negative for expression of EGFR where expression ranged from 0 to 6%. MDA-MB-231 had almost no expression of all 5 markers, with positive values ranging from 0 to 5%. MCF10A had weak positive to almost negative expression values of ER, PR, HER-2/neu, and E-cadherin, which ranged from 3 to 13%. EGFR, both surface and cytoplasmic markers, again were not expressed in MCF10A cells with an expression value of <1%. We found that ER, PR, and HER-2/neu marker expressions in 5 out of 5 cell lines were consistent with established expression patterns. EGFR and E-cadherin expression in 4 out of 5 cell lines were also consistent with established expression patterns. We have shown that flow cytometry provides quantitative data on expression patterns of important prognostic markers in breast cancer. PMID- 16271363 TI - Insulin signaling is involved in hematopoietic regulation in an insect hematopoietic organ. AB - Only a few extracellular hematopoietic factors have been identified in insects. We previously developed an in vitro culture system for the larval hematopoietic organ (HPO) of the silkworm Bombyx mori, and found that cell proliferation is linked to hemocyte discharge from the HPO. In this study, we tested hematopoietic activity of bombyxin, a peptide in the insulin family. When silkworm HPO was cultured with synthetic bombyxin-II, the number of discharged hemocytes increased in a dose-dependent manner, indicating that bombyxin promoted cell proliferation in the HPO. However, a neutralization experiment using anti-bombyxin-II antibody revealed that bombyxin is not the primary effector in larval plasma. Similarly, bovine insulin showed hematopoietic activity. Addition of molting hormone, 20 hydroxyecdysone, circumstantially enhanced the hematopoietic activity of bombyxin and insulin. Bombyxin and insulin induced phosphorylation of different sets of proteins in the HPO, suggesting that their signaling pathways are different. PMID- 16271362 TI - A high-efficiency system of natural killer cell cloning. AB - The culture of human natural killer (NK) cell clones has traditionally been a long, laborious process with an efficiency of only 1-2%. Recently, a stem cell growth medium (SCGM) has been described to expand preferentially polyclonal NK cells from peripheral blood. We have tested SCGM in a single cell sorting system and shown a 4-5 fold increase in the number of proliferating NK clones compared to standard RPMI media. The cloning efficiency was further enhanced by the provision of irradiated feeder cells derived from multiple donors combined with the addition of the anti-CD3 antibody, OKT3. The combination of SCGM, single cell sorting and these multiple optimisations enhanced NK cloning efficiency by more than tenfold to greater than 20% for short-term cultures when deriving 10(5) cells and as high as 10% for longer term cultures when deriving more than 2 x 10(6) cells. This novel system thus facilitates the generation of NK clones and allows larger scale studies of NK function that were beyond the scope of previous methodology. PMID- 16271364 TI - Lifetime associations between cannabis, use, abuse, and dependence and panic attacks in a representative sample. AB - BACKGROUND: The present investigation evaluated lifetime associations between cannabis use, abuse, and dependence and panic attacks after controlling for alcohol abuse, polysubstance use, and demographic variables. METHODS: Data for this study were obtained as part of a large statewide survey, the Colorado Social Health Survey (CSHS). Participants were contacted using randomly sampled household addresses (72% response rate) and interviews took place in participants' homes. Participants consisted of a representative sample from the Colorado general adult population (n=4745; 52% female). The Diagnostic Interview Schedule was administered to obtain diagnoses. RESULTS: After controlling for polysubstance use, alcohol abuse, and demographic variables, lifetime history of cannabis dependence, but not use or abuse, was significantly related to an increased risk of panic attacks. Additionally, among participants reporting a lifetime history of both panic attacks and cannabis use, the age of onset of panic attacks (M=19.0 years of age) was significantly earlier than for individuals with a lifetime panic attack history but no cannabis use (M=27.6 years of age). CONCLUSIONS: Structured interview data suggest lifetime cannabis dependence is significantly associated with an increased risk of panic attacks. PMID- 16271365 TI - Hepatocyte growth factor stimulates the induction of cytokine-induced neutrophil chemoattractant through the activation of NF-kappaB in rat hepatocytes. AB - BACKGROUND: Recent evidence indicates that CXC chemokines such as human interleukine-8 (IL-8) and rat cytokine-induced neutrophil chemoattractant (CINC) have mitogenic and anti-apoptotic functions in addition to their chemotactic activity toward neutrophils. In the liver, hepatocyte growth factor (HGF) is implicated as a mitogen in the regeneration of liver. However, little is known the interaction between HGF and CXC chemokines during liver injury, repair and regeneration. We hypothesized that HGF may stimulate the expression of such chemokines, which contributes to mitogenic action in liver regeneration. MATERIALS AND METHODS: Primary cultured rat hepatocytes were treated with recombinant human HGF (rhHGF), in the absence and presence of calpain inhibitor-1 (CI-1), genistein or anti-CINC-1 antibody. Levels of CINC-1 and its mRNA, tyrosine phosphorylation of HGF receptor (c-Met), the activation of transcription factor, nuclear factor-kappaB (NF-kappaB), and activities of DNA synthesis were measured. RESULTS: rhHGF enhanced the production of CINC-1 time- and dose dependently, which followed the increased levels of CINC-1 mRNA. Under the same conditions, rhHGF increased the tyrosine phosphorylation of c-Met. The electrophoretic mobility shift assay revealed that rhHGF activated the nuclear translocation of NF-kappaB and its DNA binding. Proteasome inhibitor (CI-1) blocked the NF-kappaB activation and the CINC-1 production. The tyrosine kinase inhibitor (genistein) inhibited the activities of CINC-1 production and the DNA synthesis stimulated by rhHGF. However, the treatment of anti-CINC-1 antibody had no effect on the DNA synthesis. CONCLUSIONS: These results demonstrate that rhHGF can stimulate the induction of CINC-1 gene expression through the activation of NF-kappaB via its receptor c-Met in hepatocytes. Although CINC-1 seems to be not associated with the enhancement of DNA synthesis by rhHGF, it cannot negate the possibility that CINC-1 may contribute to liver repair and regeneration. PMID- 16271367 TI - Oxygen consumption, pCO2 gradients and regional blood flow distribution in an alternative model of intestinal autotransplantation. AB - BACKGROUND: Postoperative complications after intestinal transplantation can be attributed to hypothermic storage and reperfusion injury. Our objective was to evaluate, in an alternative model of intestinal autotransplantation, the initial effects of isolated intestinal hypothermic perfusion (at 4 degrees C, IHP) on mucosal and serosal blood flow distribution and correlate these findings with other systemic and regional markers of mesenteric ischemia. In addition, we sought to obtain evidence that intestinal pCO2 measurement can be a useful method for monitoring graft perfusion and early histological changes after small bowel transplantation. MATERIALS AND METHODS: Eight dogs (23.3+/-1.1 kg) were submitted to a in situ IHP for 30 min, followed by a 180-min reperfusion period. Cardiac output, mesenteric vein, and intestinal serosal blood flows (SMVBF and SBF, ultrasonic flowprobe); intestinal mucosal-arterial pCO2 gradient (Dt-apCO2, tonometry); and O2-derived variables were evaluated. RESULTS: IHP induced a reduction in SMVBF (579+/-53 to 321+/-10 mL/min) and SBF, (44.7+/-3.2 to 29.1+/ 5.3 mL/min), and an increase in Dt-apCO2 (2+/-2.8 to 20.5+/-4.5 mm Hg). No alterations on systemic metabolic or O2-derived variables were observed. The increase of the Dt-apCO2 correlated with the grade of mucosal injury. CONCLUSION: IHP induces a proportional reduction on blood flow in all layers of the intestine, and none of the systemic markers of splanchnic ischemia predict the intestinal blood flow disturbances during the early phase of intestinal transplantation. In addition, intestinal pCO2 measurement seems to be a useful way for monitoring graft perfusion and histological changes after hypothermic ischemia and reperfusion. PMID- 16271366 TI - Effects of homocysteine and ginsenoside Rb1 on endothelial proliferation and superoxide anion production. AB - BACKGROUND: Homocysteine (Hcy) is an independent risk factor for cardiovascular disease by its multiple effects on vascular cells and throbmosis factors, which may be involved in oxidative stress mechansims. Ginsenoside Rb1, a constituent of ginseng, bears various beneficial effects on the cardiovascular system. In the present study, we investigated the effect of Hcy on endothelial proliferation and a protective effect of ginsenoside Rb1 on the action of Hcy. METHODS: We initially incubated a mouse lymph node endothelial cell line (SVEC4-10) with increasing concentrations of Hcy or for different time periods and then assessed cell proliferation by using [(3)H]-thymidine incorporation. We then incubated SVEC4-10 cells with Hcy (50 microM) for 24 h with or without Rb1 (10 microM) to examine its inhibitory effect on the proliferation. These experiments were repeated in human umbilical vein endothelial cells (HUVECs). To explore the underlying molecular mechanisms, we measured superoxide anion, a reactive oxygen species (ROS), by using dihydroethidium (DHE) staining. RESULTS: SVEC4-10 cells treated with Hcy (50, 100, and 200 microM) for 24 h significantly reduced cell proliferation by 43%, 42%, and 40%, respectively, as compared with control cells (P < 0.01). SVEC4-10 cells treated with Hcy (50 microM) for 12 and 24 h showed a significant reduction of cell proliferation (P < 0.05). In HUVECs, Hcy (50 microM) significantly reduced cell proliferation by 55% as compared with control cells (P < 0.05). In the presence of Rb1, Hcy-induced inhibition of cell proliferation was effectively blocked in both SVEC4-10 and HUVECs. Furthermore, Hcy (50 microM) significantly increased superoxide anion production by 23% in SVEC4-10 as compared with control cells (P < 0.05). However, in the presence of Rb1, Hcy increased superoxide anion production by only 8%, showing that RB1 almost completely blocked the effect of Hcy. CONCLUSION: Hcy significantly inhibits endothelial proliferation with increased production of superoxide anion, which is effectively blocked by ginsenoside Rb1. This study provides some new aspects of Hcy-induced endothelial dysfunction, and suggests a potential role of Rb1 to block Hcy action, which may have clinical applications. PMID- 16271368 TI - Effects of emodin on Ca2+ signal transduction of smooth muscle cells in multiple organ dysfunction syndrome. AB - We have made several reports on the signal transduction mechanism that emodin enhance the calcium concentrations of smooth muscle cells (SMCs) in the physiological condition by inositol [1, 4, 5]-friphosphate (IP3). The observation that IP3 concentrations in SMCs were decreased in multiple organ dysfunction syndrome (MODS) prompted us to ask whether emodin can activate SMCs to contract by way of elevating [Ca2+] and thus modulating the critical Ca2+ signal transduction pathways involved in the contraction of the SMCs in the pathological setting of MODS. To test this hypothesis, we used the rat model of MODS to explore the potential roles of emodin in Ca2+ signal transduction in the SMCs of colon in rats. ML-7 [an inhibitor of myosin light-chain kinase (MLCK)] and Calphostin C [an inhibitor of protein kinase C (PKC)] were used to observe the influence of emodin on the muscle strips and SMCs in rats after MODS. Nifedipine (an antagonist of voltage-gated Ca2+ channel), EGTA (removal of extracellular Ca2+), heparine (a specific IP3 receptor antagonist), and ryanodine were used to probe the potential mechanisms involved in emodin-mediated elevation of the global cytoplasmic Ca2+ in SMCs of colon in the rats after MODS. Our results show that emodin is capable of contract the smooth muscles of colon in rats after MODS by MLCK increasing [Ca2+] of SMCs, and by PKC enhancing the calcium sensitivity of SMCs. The mechanism by which emodin triggers elevated [Ca2+] of smooth muscles of colon in rats after MODS is likely to operate through IP3 and RyR receptors in the sarcoplasm. It is hoped that deeper insights into how emodin modulates the critical calcium signaling in SMCs might lead to the potential development of emodin in the treatment of MODS. PMID- 16271369 TI - Spatial mosaic formation through frequency-dependent selection in Mullerian mimicry complexes. AB - Although contemporary models of Mullerian mimicry have considered the movement of interfacial boundaries between two distinct mimetic forms, and even the possibility of polymorphisms in two patch systems, no model has considered how multiple forms of Mullerian mimics might evolve and be maintained over large geographical areas. A spatially explicit individual-based model for the evolution of Mullerian mimicry is presented, in which two unpalatable species are distributed over discrete cells within a regular lattice. Populations in each cell are capable of genetic drift and experience localized dispersal as well as frequency-dependent selection by predators. When each unpalatable prey species was introduced into a random cell and allowed to spread, then mimicry evolved throughout the system in the form of a spatial mosaic of phenotypes, separated by narrow "hybrid zones". The primary mechanism generating phenotypic diversity was the occasional establishment of new mutant forms in unoccupied cells and their subsequent maintenance (and spread) through frequency-dependent selection. The mean number of discrete clusters of the same morph that formed in the lattice was higher the higher the intensity of predation, and higher the lower the dispersal rate of unpalatable prey. Under certain conditions the hybrid zones moved, in a direction dependent on the curvature of their interfacial boundaries. However, the mimetic mosaics were highly stable when the intensity of predation was high and the rate of prey dispersal was low. Overall, this model highlights how a stable mosaic of different mimetic forms can evolve from a range of starting conditions through a combination of chance effects and localized frequency dependent selection. PMID- 16271370 TI - Protein kinase C inhibitor chelerythrine attenuates the morphine-induced excitatory amino acid release and reduction of the antinociceptive effect of morphine in rats injected intrathecally with pertussis toxin. AB - Neuropathic pain syndromes respond poorly to opioid treatment. In our previous studies, we found that intrathecal (i.t.) injection of pertussis toxin (PTX) produces thermal hyperalgesia, which is poorly responsive to morphine and is accompanied by an increase in cerebrospinal fluid (CSF) levels of excitatory amino acids (EAAs) and protein kinase C (PKC) activation. In the present study, rats were implanted with an i.t. catheter for drug injection and a microdialysis probe for CSF dialysate collection. On the fourth day after injection of PTX (2 microg, i.t.), there was a significant reduction in the antinociceptive effect of morphine (10 microg, i.t.) which was accompanied by an increase in levels of EAAs. Pretreatment with the PKC inhibitor, chelerythrine (25 microg, i.t.) one hour before morphine injection markedly inhibited both effects. These results suggest that, in PTX-treated rats, PKC plays an important role in inhibiting the morphine-induced spinal EAA release, which might be related to the reduced antinociceptive effect of morphine. PMID- 16271372 TI - Distribution of silver in mussels and oysters along the French coasts: data from the national monitoring program. PMID- 16271371 TI - Implication of glucocorticoid in anti-inflammatory effects of Ro5-4864 in mouse pleurisy induced by carrageenan. AB - Mouse pleurisy induced by carrageenan is used to determine the mechanism of anti inflammatory action of 7-chloro-5-(4-chlorophenyl)-1,3-dihydro-1-methyl-2-H-1,4 benzodiazepin-2 (Ro5-4864). Pre-treatment with Ro5-4864 inhibits different inflammatory parameters, such as neutrophil influx, MPO activity and NO levels in the early phase (4 h), as well as mononuclear cells and ADA activity in the late phase (48 h) of pleurisy. dl-Aminoglutethimide, inhibitor of steroidal synthesis, reverted the effect of Ro5-4864 on these different inflammatory parameters. Our results suggest that anti-inflammatory action of Ro5-4864 may be partly due to its capacity to inhibit leukocyte migration, as well as leukocyte activation and formation of NO by a mechanism dependent on glucocorticoids. PMID- 16271373 TI - Cell and tissue biomarkers in mussel, and histopathology in hake and anchovy from Bay of Biscay after the Prestige oil spill (Monitoring Campaign 2003). AB - In order to assess the biological effects of the Prestige oil spill (POS), mussels (Mytilus galloprovincialis), European hake (Merluccius merluccius) and European anchovy (Engraulis encrasicolus) were sampled between April and September 2003 in various geographical areas of Bay of Biscay: Galicia, Central Cantabrian and East Cantabrian. In mussels, several cell and tissue biomarkers were measured: peroxisome proliferation as induction of acyl-CoA oxidase (AOX) activity, lysosomal responses as changes in the structure (lysosomal volume density, V(V(L)), surface-to-volume ratio, S/V(L), and numerical density, N(V(L))) and in membrane stability (labilization period, LP), cell type replacement as relative proportion of basophilic cells (volume density of basophilic cells, V(V(BAS))) in digestive gland epithelium, and changes in the morphology of digestive alveoli as mean luminal radius to mean epithelial thickness (MLR/MET). Additionally, flesh condition index (FCI) and gonad index (GI) were measured as supporting parameters. In hake and anchovy, liver histopathology was examined to determine the prevalence of parasites, melanomacrophage centers, non-specific lesions (inflammatory changes, atrophy, necrosis, apoptosis), early non-neoplastic toxicopathic lesions (i.e., hepatocellular nuclear polymorphism), foci of cellular alteration, benign and malignant neoplasms. In mussels, AOX induction was noticeable in April except in Galicia. LP values were low in all the geographical areas studied, indicating disturbed health, especially in Galicia. Alike, lysosomal enlargement was observed in most stations as shown by the extremely low S/V(L) values. V(V(BAS)) and MLR/MET values were markedly high. Overall, employed biomarkers detected exposure to toxic chemicals and disturbed health in mussels from Bay of Biscay, with impact decreasing from April to September. Although hepatocellular nuclear polymorphism and nematode parasitization in fish liver were remarkably prominent in some areas, they cannot be hitherto related to POS, since we lack historical data to determine whether the prevalences found were normal or significantly raised after POS. PMID- 16271374 TI - Effect of copper on fertilization success in the reef coral Acropora surculosa. PMID- 16271375 TI - Acylated farnesyl diglycosides from Guioa crenulata. AB - Chemical investigation of the methanol extract of the leaves of Caledonian Guioa crenulata led to the isolation and characterisation of four farnesyl diglycosides, crenulatosides A, B, C and D, along with three known flavonol glycosides and one known trimeric proanthocyanidin possessing a doubly linked structure. The structures of these compounds were determined on the basis of spectroscopic studies and chemical evidence. The ethanol and ethyl acetate extracts of the leaves exhibited no cytotoxic activity and no inhibition of acetylcholinesterase. PMID- 16271376 TI - Tricalysiosides H-O: Ent-kaurane glucosides from the leaves of Tricalysia dubia. AB - Eight ent-kaurane glucosides, named tricalysiosides H-O, were isolated from Tricalysia dubia. Tricalysioside H possessed a hydroxyl group at the 1-position, to which the glucose moiety was attached. The structure was first elucidated by means of spectroscopic data analysis and finally confirmed by X-ray crystallography. Since acid hydrolysis of 1 gave D-glucose, the aglycone was proved to have an enantio-kaurane type skeleton. The structures of tricalysiosides I-O were mainly elucidated from analysis of spectroscopic evidence. PMID- 16271377 TI - Lack of WHV integration nearby N-myc2 and in the downstream b3n and win loci in a considerable fraction of liver tumors with activated N-myc2 from naturally infected wild woodchucks. AB - In liver tumors induced by chronic WHV infection in the WHV/woodchuck model of HBV infection, activation of genes of the myc family by WHV insertion has been well documented. Several studies have shown that N-myc2 is by far the most frequently involved, and in most cases, its transcriptional activation is due to WHV insertion nearby the gene. N-myc2 has been shown to be also activated by WHV insertion in two downstream loci, b3n and win. Although the extent of insertion in these latter loci in woodchuck tumors has not been investigated, their discovery has led to the notion that therein WHV insertion accounts for N-myc2 activation in the remaining tumors expressing the proto-oncogene in absence of any detectable alteration nearby the gene, a notion remained unproved and not further investigated yet. In the majority of cases, the above observations were derived from tumors developed in colony born laboratory bred woodchucks experimentally infected with standardized viral inocula, mostly of the same lineage. In the present work, we investigated a survey of liver tumors naturally developed in wild woodchucks with naturally acquired chronic WHV infection. Tumors had histological features of well to moderately differentiated HCCs. In most animals, multiple tumor nodules were observed; in the great majority of cases, they were shown to be independent tumors because their WHV integration patterns were not clonally related. 53 independent tumors were investigated for N myc activation and WHV integration nearby N-myc genes and in the b3n and win loci. Comparison of our results with data from previous studies revealed that, in tumors from naturally infected wild woodchucks, the frequency of WHV integration nearby N-myc2 has a tendency to be lower and, in addition, N-myc2 activation is due to WHV integration nearby the gene significantly less frequently than in tumors from experimentally infected colony born animals (12/28, 43% vs. 15/20, 75%, P = 0.0397). These findings are likely related to the less uniform conditions as to infecting virus and host genetic background in naturally infected wild woodchucks with respect to experimentally infected colony born woodchucks and suggest that viral and/or host factors may influence the site of viral insertion finally detected in overt tumors. In addition, more than one third (11/28, 39%) tumors with activated N-myc2 transcription did not show rearrangement either nearby the gene, or in b3n or in win. These findings challenge the notion that integration in the downstream b3n and win loci is responsible for N-myc2 activation in tumors lacking insertion nearby N-myc2 and suggest that in a considerable fraction of liver tumors, at least from wild woodchucks, N-myc2 activation might be due either to WHV integration in further regions of the N-myc2 chromosomal domain or to other mechanisms related or unrelated to viral insertion. PMID- 16271378 TI - Recombinant influenza A viruses harboring optimized dicistronic NA segment with an extended native 5' terminal sequence: induction of heterospecific B and T cell responses in mice. AB - We generated novel recombinant influenza A viruses (vNA38) harboring dicistronic NA segments with an extended native 5' terminal sequence of 70 nucleotides comprised of the last 42 nucleotides of the NA ORF and the 5' noncoding region (5' NCR). vNA38 viruses replicated stably and more efficiently than vNA35 viruses with a dicistronic NA segment comprised of the native 5' NCR only, that we described previously (Vieira Machado, A., Naffakh, N., van der Werf, S., Escriou, N., 2003. Expression of a foreign gene by stable recombinant influenza viruses harboring a dicistronic genomic segment with an internal promoter. Virology 313, 235-249). In addition, vNA38 viruses drove the expression of higher levels of encoded heterologous proteins than corresponding vNA35 viruses, both in cell culture and in the pulmonary tissue of infected mice. These data demonstrate that a sequence overlapping 5' coding and noncoding regions of the NA segment determines efficient replication and/or propagation of the vRNA. Intranasal immunization of mice with live vNA38 viruses induced B and T cell responses specific for the heterologous protein expressed, establishing the usefulness of such recombinant influenza viruses with a dicistronic segment for the development of live bivalent vaccines. PMID- 16271379 TI - 1,2,4-trichlorobenzene marine risk assessment with special emphasis on the Osparcom region North Sea. AB - A risk assessment on 1,2,4-trichlorobenzene was carried out specifically for the marine environment according to the methodology laid down in the EU Risk Assessment Regulation 1488/94 and the Guidance Documents of the EU Existing Substances Regulation 793/93. The study consists of the collection and evaluation of data on effects and environmental concentrations from analytical monitoring programs in large rivers and estuaries in the North Sea area. The risk is indicated by comparing the predicted environmental concentration (PEC) with the predicted no-effect concentrations (PNEC) for the marine aquatic environment. A PNECwater) value of 0.3 microg/l and a PNECsed value of 38 microg/kgdw were derived from the results of toxicological studies in organisms representing three trophic levels, i.e. aquatic plants, invertebrates and fish. Based on monitoring data two situations are distinguished: a typical case and a worst case with a PECwater of <0.047 and 0.1 microg/l, respectively, and a PECsed of 40 and 90 microg/kgdw, respectively. The calculated PEC/PNEC ratios were 0.16 and 0.3 for water and 1 and 2.4 for sediment, respectively. It was concluded that no risks are expected for aquatic organisms. Based on the combination of worst-case assumptions risks to benthic organisms could not be fully excluded, but since all open uses of 1,2,4-trichlorobenzene will be ended following the EU risk assessment outcome of 2001 any potential risk is expected to be reduced accordingly. 1,2,4-trichlorobenzene is not considered toxic according to the EU criteria and the available data on persistence of 1,2,4-trichlorobenzene indicate a half-life in water of a few days and a significant biodegradation potential. The bioaccumulation potential is low to moderate with most BCF ratios for fish ranging from 600 to 1400 and one highest of 2020. Based on an extensive evaluation of persistence, biodegradation and bioaccumulation data it is concluded that 1,2,4-trichlorobenzene is not a PBT, since it does not fulfill any of the EU criteria. Biomagnification in the food chain is not expected due to the relatively high elimination rate constants. PMID- 16271381 TI - Mercury concentrations of fish, river water, and sediment in the Rio Ramis-Lake Titicaca watershed, Peru. AB - This study reports the first set of data on the concentration of mercury in muscle tissue of several varieties of fish from Lake Titicaca, including the pejerrey (Basilichthyes bonariensis), the carachi (Orestias), and 2 types of indigenous catfish (Trichomycterus). Approximately 27% of the pejerrey and 75% of the carachi exceeded the US EPA fish tissue-based water quality criterion level of 0.30 microg g(-1). Mercury levels of pejerrey increased with fish size, although this relationship was less apparent for the smaller carachi. The pejerrey and carachi are important food fish for local residents. A synoptic sampling of the Rio Ramis--the largest tributary to Lake Titicaca--was conducted in an attempt to determine if mercury releases from artisanal gold mining could be an important source of Hg contamination to Lake Titicaca. Although highly elevated concentrations of Hg and other heavy metals were documented in headwater streams near the mining centers of La Rinconada and Cecilia, the quantity of Hg entering Lake Titicaca that could be attributed to mining in the Ramis watershed was below the quantifiable limit in our July 2002 study. This does not diminish the localized threat to mercury exposure for the artisanal gold miners themselves, as well as their families. Further studies of mercury dynamics in Lake Titicaca are recommended, as well as in the rivers draining into the lake. It is probable that most of the downgradient transport of Hg and other trace metals from the headwater mining centers occurs as suspended sediment during seasonal periods of high-flow. PMID- 16271380 TI - Potential impacts of water injection dredging on water quality and ecotoxicity in Limehouse Basin, River Thames, SE England, UK. AB - The use of water injection dredging (WID) is increasing in the UK's inland waterways and marinas. Jets of water are injected under low pressure directly into bottom sediment creating a turbulent water-sediment mixture that flows under the influence of gravity. Many of these sediments are highly contaminated and little is known of the effects of contaminant release on water quality or the risk to biota living in both the sediment and the water column. Sediment cores were collected from Limehouse Basin, a proposed WID site in SE England and current sediment toxicity was assessed using a number of techniques. Comparison of metal data to US sediment quality guidelines indicated intermediate levels of toxicity while, calculation of acid volatile sulphide to simultaneously extracted metal ratios underestimated the potential toxicity to sediment dwelling organisms. In contrast, porewater ammonia concentrations were in excess of all published ecotoxicological guidelines and indicate serious risk to biota. Re suspension experiments were used to mimic the effects of WID on overlying water quality and ecotoxicity tests were carried out on elutriates using Daphnia magna to examine the impacts on biota. Concentrations of a range of metals in the elutriates predict that adverse biological effects would be observed during WID, however only 10% of the elutriate samples caused an adverse effect on Daphnia. Limehouse Basin is a complex aquatic environment receiving predominantly fresh waters while the sediments have high porewater chloride concentrations reminiscent of previous tidal inputs to the basin, making the choice of test organism problematic. PMID- 16271382 TI - Populations of exopolysaccharide-producing cyanobacteria and diatoms in the mucilaginous benthic aggregates of the Tyrrhenian Sea (Tuscan Archipelago). AB - The microalgal community of benthic mucilages sampled in various sites of the Tuscan Archipelago during the period May 1999-July 2002, was always dominated by diatoms. Synedra, Licmophora and Navicula were the most frequently found genera, their presence having been observed within all the period under study; other ten genera of diatoms were found in the benthic mucilages, but their presence was dependent on the month and the year of observation. As concerns the cyanobacterial community, the filamentous genera Leptolyngbya, Lyngbya and Rivularia were the most frequently found, independently on the period and the year of samplings, while other genera, namely Oscillatoria, Symploca, Aphanocapsa and Gloeocapsa, were less frequently observed. During the period under study, the highest number of different genera of diatoms and cyanobacteria in the mucilaginous aggregates was observed during summer periods, particularly in June and July. The microscopic observation of the samples demonstrated that only diatoms belonging to the genera Navicula, Amphora, Cylindrotheca and Pseudo nitzschia possessed outermost polysaccharidic investments, the cells of the other genera being devoid of sheaths or capsules. On the other side, all the observed cyanobacteria, with the exception of Oscillatoria strains, were characterized by the presence of outermost polysaccharidic investments, most frequently in the form of sheaths. However, when the microalgal and cyanobacterial isolates were cultured under laboratory conditions, the release of polysaccharidic material into the culture medium was observed only in a few cases and always in very small amounts. The observations done on the natural samples and the results obtained with the cultures of the microalgae and cyanobacteria isolated from the benthic aggregates suggest that only diatoms have given some contribution to the formation of the polysaccharidic material constituting the matrix of the mucilages, whereas the role of cyanobacteria in the benthic aggregates seems to be related to their capability to create biofilms on solid surfaces, constituting the first step in the colonization of the benthic surfaces by the microbial community. PMID- 16271383 TI - Exposure of garden birds to aflatoxins in Britain. AB - Aflatoxins are potent biological toxins that have been shown to exert a range of acute and chronic pathological effects. Multiple mortality events of waterfowl caused by acute aflatoxicosis have been documented in the USA. However, international concern has recently been expressed regarding the potential effects of chronic exposure of wildlife species to low levels of dietary aflatoxin. This study documents for the first time the presence of hepatic aflatoxin residues in British wild birds: two passerine species, the house sparrow (Passer domesticus) and greenfinch (Carduelis chloris). Further research is required to investigate the source of the dietary aflatoxins and their pathological significance, if any, for wild birds in Britain. PMID- 16271384 TI - Phase II trial of imatinib mesylate in patients with recurrent platinum- and taxane-resistant epithelial ovarian and primary peritoneal cancers. AB - OBJECTIVES: To evaluate the efficacy and tolerability of imatinib mesylate (Gleevec; Novartis Pharmaceuticals, Basel, Switzerland) in patients with recurrent ovarian and primary peritoneal cancer. METHODS: This was an open-label, single-institution phase II trial. Patients were eligible if they had measurable platinum/taxane-resistant disease, received 2-4 prior treatment regimens, and over-expressed at least one imatinib target (c-Kit, PDGFR-beta, or c-Abl) by immunohistochemistry. Imatinib was administered orally at 600 mg daily for 6 weeks (one course) and was repeated in the absence of measurable progression. RESULTS: Sixteen enrolled patients were evaluable for toxicity and 12 for response. The median number of prior treatments was 4. A total of 29 courses were initiated. No complete or partial responses were documented during a median follow-up of 6.6 months. However, 4 (33%) of the 12 evaluable patients had stable disease lasting 3.8, 6.4, 7.5, and 8+ months. Expression of PDGFR-beta and c-Abl was seen in 15 (94%) and c-Kit in 8 (50%) patients' tumors. There was no relationship between best response (stable disease) and target expression. Adverse events were uncommon, with fatigue and nausea/vomiting being reported in 34% and 31% of cycles, respectively. Two patients underwent dose reduction for rash and edema (n = 1) and grade 3 neutropenia (n = 1). No grade 4 toxicity was observed. CONCLUSION: Imatinib mesylate was well tolerated but did not produce clinical responses in patients with previously treated metastatic ovarian and primary peritoneal carcinoma. PMID- 16271385 TI - Argonaute2 cleaves the anti-guide strand of siRNA during RISC activation. AB - The mRNA-cleavage step of RNA interference is mediated by an endonuclease, Argonaute2 (Ago2), within the RNA-induced silencing complex (RISC). Ago2 uses one strand of the small interfering (si) RNA duplex as a guide to find messenger RNAs containing complementary sequences and cleaves the phosphodiester backbone at a specific site measured from the guide strand's 5' end. Here, we show that both strands of siRNA get loaded onto Ago2 protein in Drosophila S2 cell extracts. The anti-guide strand behaves as a RISC substrate and is cleaved by Ago2. This cleavage event is important for the removal of the anti-guide strand from Ago2 protein and activation of RISC. PMID- 16271386 TI - Passenger-strand cleavage facilitates assembly of siRNA into Ago2-containing RNAi enzyme complexes. AB - In the Drosophila and mammalian RNA interference pathways, siRNAs direct the protein Argonaute2 (Ago2) to cleave corresponding mRNA targets, silencing their expression. Ago2 is the catalytic component of the RNAi enzyme complex, RISC. For each siRNA duplex, only one strand, the guide, is assembled into the active RISC; the other strand, the passenger, is destroyed. An ATP-dependent helicase has been proposed first to separate the two siRNA strands, then the resulting single stranded guide is thought to bind Ago2. Here, we show that Ago2 instead directly receives the double-stranded siRNA from the RISC assembly machinery. Ago2 then cleaves the siRNA passenger strand, thereby liberating the single-stranded guide. For siRNAs, virtually all RISC is assembled through this cleavage-assisted mechanism. In contrast, passenger-strand cleavage is not important for the incorporation of miRNAs that derive from mismatched duplexes. PMID- 16271387 TI - Human RISC couples microRNA biogenesis and posttranscriptional gene silencing. AB - RNA interference is implemented through the action of the RNA-induced silencing complex (RISC). Although Argonaute2 has been identified as the catalytic center of RISC, the RISC polypeptide composition and assembly using short interfering RNA (siRNA) duplexes has remained elusive. Here we show that RISC is composed of Dicer, the double-stranded RNA binding protein TRBP, and Argonaute2. We demonstrate that this complex can cleave target RNA using precursor microRNA (pre miRNA) hairpin as the source of siRNA. Although RISC can also utilize duplex siRNA, it displays a nearly 10-fold greater activity using the pre-miRNA Dicer substrate. RISC distinguishes the guide strand of the siRNA from the passenger strand and specifically incorporates the guide strand. Importantly, ATP is not required for miRNA processing, RISC assembly, or multiple rounds of target-RNA cleavage. These results define the composition of RISC and demonstrate that miRNA processing and target-RNA cleavage are coupled. PMID- 16271388 TI - Evaluating the effects of protection on two benthic habitats at Tavolara-Punta Coda Cavallo MPA (North-East Sardinia, Italy). AB - In this paper, two benthic habitats have been investigated in a fully protected site and two control sites at the Tavolara-Punta Coda Cavallo marine protected area (MPA) (NE Sardinia). Overall, a protection effect on the benthic assemblages was highlighted in the shallow subtidal (5 m deep) rather than at intertidal algal turf habitat. Structure of assemblages at the shallow subtidal habitat is different in the fully protected site vs. controls, while this was not true for the intertidal habitat. At the subtidal, this finding is probably linked to indirect effects due to an increase of consumers in the protected site (e.g., sea urchins), while the lack of direct impacts in the intertidal at control sites is indicative of very similar assemblages. Cover of encrusting algae was significantly higher at the subtidal protected site suggesting a possible higher grazing pressure. Possible causes underlying the inconsistency of results obtained between habitats include the possibility that trophic cascade effects have a different influence at different heights on the shore. The need to estimate the interconnection among benthic habitats through trophic links is also highlighted to provide an estimate of the vulnerability to protection of various habitats. PMID- 16271389 TI - The narrative structure of psychiatric reports. AB - The present contribution illustrates the findings of a research about the narrative structure of psychiatric expertise. We have analysed a sample of nine expertises, using the methodology proposed by Roland Barthes in his book S/Z, in which he applies it to Honore de Balzac's "Sarrasine". Barthes suggests that every narrative is characterized by the presence of five codes (hermeneutical, proairetic, semantic, symbolic and referential): we have searched for them in the expertises, but we have also observed the presence of many lapsus calami. As for our results, we have interpreted the massive presence of symbolic code in an expertise as a proof of the elaboration of the horror elicited by crime in the expert's narrative and, on the other side, the presence of referential code as the proof of the incapacity to treat such feeling and the necessity to defend himself from it. In this vision, lapsuses arise when the expert is astonished by the horror of the crime, and does not succeed in elaborating it either by using symbolic code or through his cultural resources (referential code). Finally, some reflections are made upon the difficulty for the voice of the defendant to be heard in the texts written about him. PMID- 16271390 TI - Long-term study of environmental levels of dioxins and furans in the vicinity of a municipal solid waste incinerator. AB - From 1975 to 2004, a municipal solid waste incinerator (MSWI) was operating in Montcada (Barcelona, Catalonia, Spain). Because of the potential health risks derived from emission of pollutants by the facility, especially polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzofurans (PCDFs), a long-term monitoring program focused on measuring the environmental levels of PCDD/Fs near the facility, and to assess the health risks for the population living in the neighborhood, was established between 1996 and 2002. A total number of 111 soil and 121 herbage samples were analyzed for PCDD/Fs during this period. Human health risks for the individuals living near the MSWI (500 and 1,000 m) were also assessed before (1998) and after modernization (2000) of the facility. It included PCDD/F inhalation, dermal contact, soil and dust ingestion, and food intake. All these data are here summarized. The environmental levels of PCDD/Fs showed that the MSWI was not the main responsible of the atmospheric pollution by these compounds. In turn, human health risks for the population living in the vicinity of the facility after introduction of a modern technology were negligible in comparison with the dietary PCDD/F exposure. PMID- 16271391 TI - NK-lysin of channel catfish: gene triplication, sequence variation, and expression analysis. AB - Antimicrobial peptides (AMPs) are important components of the host innate immune response against microbial invasion. In addition to the previously known four classes of antimicrobial peptides, a fifth class of antimicrobial peptides has been recently identified to include NK-lysins that have a globular three dimensional structure and are larger with 74-78 amino acid residues. NK-lysin has been shown to harbor antimicrobial activities against a wide spectrum of microorganisms including bacteria, fungi, protozoa, and parasites. To date, NK lysin genes have been reported from only a limited number of organisms. We previously identified a NK-lysin cDNA in channel catfish. Here we report the identification of two novel types of NK-lysin transcripts in channel catfish. Altogether, three distinct NK-lysin transcripts exist in channel catfish. In this work, their encoding genes were identified, sequenced, and characterized. We provide strong evidence that the catfish NK-lysin gene is tripled in the same genomic neighborhood. All three catfish NK-lysin genes are present in the same genomic region and are tightly linked on the same chromosome, as the same BAC clones harbor all three copies of the NK-lysin genes. All three NK-lysin genes are expressed, but exhibit distinct expression profiles in various tissues. In spite of the existence of a single copy of NK-lysin gene in the human genome, and only a single hit from the pufferfish genome, there are two tripled clusters of NK-lysin genes on chromosome 17 of zebrafish in addition to one more copy on its chromosome 5. The similarity in the genomic arrangement of the tripled NK-lysin genes in channel catfish and zebrafish suggest similar evolution of NK-lysin genes. PMID- 16271392 TI - A complement receptor for opsonized immune complexes on erythrocytes from Oncorhynchus mykiss but not Ictalarus punctatus. AB - Immune complexes activate the classical pathway of complement resulting in the covalent deposition of fragments of the third (C3b) and fourth (C4b) components of complement, thus opsonizing the complexes for uptake by CD35 found on human erythrocytes. The complexes are then transported to and cleared from the circulation by the reticuloendothelial system. It has been shown that rainbow trout can remove immune complexes from the circulation in a complement-dependent manner similar to that found in the human. However, the cell or cell types involved have not been identified. The purpose of this study was to investigate whether a complement-dependent immune adherence receptor is expressed on erythrocytes from the rainbow trout (Oncorhynchus mykiss) and the channel catfish (Ictalarus punctatus). Coating fluorescent microparticles with BSA, and then binding them to anti-BSA created an artificial immune complex that was incubated with normal fish serum, normal human serum or EDTA-treated serum. The complement coated immune complexes were then incubated with either fish or human erythrocytes and analyzed for binding by flow cytometry and further visualized by fluorescence microscopy. Our results indicate that erythrocytes from rainbow trout are capable of binding immune complexes when pretreated with serum from either the trout or human, but not when pretreated with serum containing EDTA. By contrast, erythrocytes from the channel catfish did not bind immune complexes pretreated with autologous or human serum. These data suggest that differences exist in receptor distribution between two closely related species of fish, and a potentially homologous relationship in receptor expression, and possibility function, exist between two highly divergent species. PMID- 16271393 TI - Identification and molecular characterization of a peritrophin-like protein from fleshy prawn (Fenneropenaeus chinensis). AB - Peritrophin, one of the components of the peritrophic matrix, was first isolated from the intestine of insects. It is thought to protect insects from invasion of microorganisms and to stimulate digestion of food. Peritrophin-like proteins have also been found in crustaceans, as a component of the egg layer. In this study, one fragment of the peritrophin-like gene was obtained from fleshy prawn (Chinese shrimp) (Fenneropenaeus chinensis) by panning the T7 phage display library constructed with the shrimp hemocyte cDNA. The total sequence of the peritrophin cDNA was cloned by modified SMART cDNA and LD-PCR methods. The full cDNA is 1048bp and the deduced protein is composed of 274 amino acids, including 21 amino acid signal peptide, and four peritrophin A domains and the latter three forming three chitin-binding domains. Similarity analysis results showed that the peritrophin-like protein from F. chinensis has significant similarities with peritrophin-like and cortical rod proteins from other shrimp. It was inducing expression in hemocytes, heart, stomach, gut, and gills of the infected shrimp, and constitutive expression in the ovaries. No expression signal was detected in the hepatopancreas of either infected or noninfected shrimp. The recombinant peritrophin-like protein has the activity of binding Gram-negative bacteria and strong binding activity to chitin. Therefore, the bacteria and chitin binding activities of the peritrophin-like protein suggest that it may plays a role in immune defense and other physiological resposes. PMID- 16271395 TI - Thermodynamic and spectroscopic studies of copper (II) complexes with three bis(amide) ligands derived from L-tartaric acid. AB - The interactions of three bis(amide) ligands derived from tartaric acid with copper (II) were investigated in aqueous solution by a combination of potentiometry, UV-vis spectrophotometry, electron paramagnetic resonance (EPR), and mass spectrometry. The formation constants of the complexes were measured and their relative structures were reported. The sites of complexation of these ligands are investigated based mostly on their electronic and EPR spectra and on the comparison with the behaviour of some analog compounds. PMID- 16271394 TI - Model studies of cholesterol and ascorbate oxidation by copper complexes: relevance to Alzheimer's disease beta-amyloid metallochemistry. AB - The neurotoxicity of the amyloid-beta peptide (Abeta) is causally linked to Alzheimer's disease (AD) and may be related to the redox chemistry associated with its interactions with copper ions and cholesterol in brain tissue. We have used density functional theory (DFT) calculations to study the mechanism controlling the Abeta/Cu catalyzed oxidation reactions of cholesterol and ascorbate using a model system. The computed results based on a binuclear Cu complex predict that oxidation of cholesterol (yielding 4-cholesten-3-one as a specific product) proceeds at a slow rate when catalyzed by a Abeta/Cu(II)|His |Cu(II)/Abeta) aggregate. The computed results also suggest that monomeric Abeta/Cu(II) is not able to oxidize cholesterol. DFT also predicted that Abeta will cross-link via covalent dityrosine formation during the oxidation of ascorbate but not during the oxidation of cholesterol. Experimental data were consistent with these predictions. PMID- 16271397 TI - For debate: is disinfection of specimens, which may contain unknown or bio terrorist organisms, essential before electron microscopic examination? AB - Severe and unexpected infections may be due to bioterrorism (BT), or the natural emergence of novel micro-organisms. Whatever the cause, there will be an urgent need to identify it for several reasons: defusing public anxiety, providing logical management and countermeasures, etc. For viruses and some bacteria, electron microscopy provides the fastest answers and identifying the cause quickly may eliminate BT. In the interests of safety, many would recommend that the specimen be disinfected at some stage before examination, but with unknown agents, however, reliable disinfection cannot be guaranteed and attempting to do so may also impair structure sufficiently to make recognition difficult or even impossible. As a basis for debate, this paper discusses the pros and cons of disinfecting such specimens. PMID- 16271396 TI - Sero-epidemiology of hepatitis B infection in pregnant women in Oman, Qatar and the United Arab Emirates. AB - OBJECTIVE: The aim of this cross-sectional study was to evaluate the prevalence of hepatitis B 'e' antigen (HBeAg) and hepatitis B surface antigen (HBsAg) in pregnant women in the Gulf States. METHODS: A total of 1710 pregnant women aged 15-45 years were enrolled in June-July 2000 in nine centres across Oman, Qatar and Unite Arab Emirates. The percentage of HBsAg-positive and HBeAg-positive individuals was determined. RESULTS: Serology results were available for 1694 women. A total of 7.1% of the women in Oman, 1% in Qatar and 1.5% in UAE were HBsAg-positive. Three (0.5%) women in Oman were HBeAg-positive. Risk factors identified for being HBsAg-positive were younger age, being a national (i.e. not an expatriate) and residing outside the city. CONCLUSIONS: Results from this study have shown that hepatitis B virus (HBV) prevalence in pregnant women is of intermediate endemicity in Oman and of low endemicity in Qatar and UAE. Thus, universal vaccination of all female adolescents before potential pregnancy is recommended in Oman in order to prevent perinatal infection, as well as to minimize horizontal transmission of the HBV in the community. PMID- 16271398 TI - Contribution from neurophysiological and psychological methods to the study of motor imagery. AB - This paper reviews studies on neurophysiological and behavioral methods used to evaluate motor imagery accuracy. These methods can be used when performed in the field and are based on recordings of peripheral indices such as autonomic nervous system or electromyographic activities, mental chronometry and psychological tests. Providing physiological signs that correlate to these types of mental processes may be considered an objective approach for motor imagery analysis. However, although autonomic nervous system activity recording has been shown to match motor imagery in real time, to evaluate its accuracy qualitatively and the individual ability to form mental images, the relationship between physiological responses and mental processes remains an inference. Moreover, electromyographic recordings may be associated with postural control data, but due to inconsistent results, they remain insufficient to solely evaluate motor imagery accuracy. Other techniques traditionally used in psychology and cognitive psychology are questionnaires, "debriefing" with subjects and mental chronometry. Although such methods lead to interesting results, there remains an important part of subjectivity as subjects perform an auto-evaluation of motor imagery accuracy. Similarly, mental chronometry gives information on the ability to preserve temporal organization of movement but does not allow the evaluation of the vividness of mental images. Thus, several methods should be combined to analyze motor imagery accuracy in greater detail. Neurophysiological recordings cannot therefore be considered an alternative but rather a complementary technique to behavioral and psychological methods. The advantages and inconvenient of each technique and the hypotheses that could be tested are discussed. PMID- 16271399 TI - Cytoskeletal, synaptic, and nuclear protein changes associated with rat interface organotypic hippocampal slice culture development. AB - Although organotypic hippocampal slice cultures (OHSCs) are used to study function within the hippocampus, the effect of maintenance in vitro upon protein expression is not fully understood. Therefore, we examined developmental changes in cultures prepared from P8 rats and maintained on porous membranes between medium and atmosphere. Between 7 and 28 days following explantation, altered hippocampal morphology could not be detected despite a significant decrease in both MAP-2c and a mid-range tau isoform by 21 DIV. During the same period, lower GFAP expression was observed, and GFAP labeling suggested a migration of astrocytes to the slice-atmosphere interface. In contrast, levels of the synaptic proteins synaptophysin and PSD-95 were significantly increased, but GAP-43 was not. The preservation of myelinated axons and synapses, along with glial and endothelial cells, was confirmed by ultrastructural analysis. Furthermore, intranuclear inclusion bodies, which are associated with normal aging in vivo, were detected in the CA1 pyramidal layer in cultures older than 14 DIV. When OHSCs were maintained for approximately 3, 4, and 10 weeks, a rise and then fall in the expression of synaptophysin and, especially, PSD-95 were found, and the biphasic trend paralleled by significant changes in Schaffer collateral-evoked excitatory post-synaptic potentials from CA1 neurons. Our data not only describe changes in cytoskeletal, synaptic, and nuclear proteins related to the maintenance of interface OHSCs, but also emphasize the potential of the model for the study of age-related phenomena within the hippocampus. PMID- 16271400 TI - Vaccination with a MHC class II peptide in Alum and inactive pertussis strongly ameliorates clinical MG in C57BL/6 mice. AB - We have investigated the efficacy of immunization against peptides from predisposing MHC class II molecules in human-compatible adjuvants for ameliorating experimental autoimmune myasthenia gravis (EAMG). C57BL/6 mice were immunized three times with the peptide I-Abetab62-76 in Alum+killed pertussis organisms (PT) prior to two injections with tAChR. The treatment greatly reduced the occurrence and severity of clinical MG relative to controls that received saline/Alum+PT or none. It also reduced antibody and T-cell responses against tAChR. The results have important implications for the possible immunotherapy of MG by targeting disease-associated MHC. PMID- 16271401 TI - Measurement of antibodies to avian influenza virus A(H7N7) in humans by hemagglutination inhibition test. AB - During the epizootic of highly pathogenic avian influenza A(H7N7) in 2003 in The Netherlands, RT-PCR and culture confirmed infection was detected in 89 persons who were ill. A modified hemagglutination inhibition (HI) test using horse erythrocytes and 2 hemagglutinating units of virus was applied to assess retrospectively the extent of human (subclinical) infection. Validation of the HI test with sera from 34 RT-PCR and culture confirmed A(H7) infected persons and sera from 100 persons from a human influenza vaccine trial in autumn 2002 showed that this HI-test had a sensitivity of 85% and a specificity of 100% when using a cut-off titer of > or =10. Using this cut-off value, A(H7) specific antibodies were detected in 49% of 508 persons exposed to poultry and in 64% of 63 persons exposed to A(H7) infected persons. Correlation of seropositivity with the occurrence of eye symptoms in exposed persons who had not received antiviral prophylaxis and of reduced seropositivity with taking antiviral prophylaxis provided further evidence that the A(H7) HI antibody titers were real. In conclusion, by applying an HI-test using horse erythrocytes human antibodies against the avian A(H7N7) virus were detected with high sensitivity and specificity in an unexpectedly high proportion of exposed persons. PMID- 16271402 TI - Is the vomeronasal system really specialized for detecting pheromones? AB - Many academics, clinicians and lay readers of science incorrectly assume that vomeronasal processing is equivalent to pheromone processing. We review the abundant data concerning the roles of both the olfactory and the vomeronasal systems in the processing of both pheromones and other odorants, demonstrating that this "equivalency hypothesis" is untenable. This conclusion has important implications for the design and interpretation of experiments examining vomeronasal and olfactory system function. We describe some of the problems that arise from assuming that this equivalency holds. Two alternative hypotheses have been offered, but the available data do not enable us to accept or reject either one. Perhaps no single functional description can adequately characterize the role of the vomeronasal system. PMID- 16271403 TI - A role for circuit homeostasis in adult neurogenesis. AB - Insertion of new neurons into adult neural circuits could either promote or impair circuit function, depending on whether homeostatic mechanisms are in place to regulate the resulting changes in neural activity. In the hippocampus (a mammalian forebrain structure important in aspects of memory and mood) several lines of behavioral evidence suggest important adaptive roles for adult-generated neurons, indicating that there could be mechanisms to control the potentially adverse increase in excitation associated with new cells. Here, we delineate behavioral and computational models for the role of circuit homeostasis in enabling neuron insertion to modulate hippocampal function adaptively, and we describe molecular and cellular mechanisms for implementing this circuit-level adaptive regulation of hippocampal activity. PMID- 16271405 TI - Partitioning of membrane molecules between raft and non-raft domains: insights from model-membrane studies. AB - The special physical and functional properties ascribed to lipid rafts in biological membranes reflect their distinctive organization and composition, properties that are hypothesized to rest in part on the differential partitioning of various membrane components between liquid-ordered and liquid-disordered lipid environments. This review describes the principles and findings of recently developed methods to monitor the partitioning of membrane proteins and lipids between liquid-ordered and liquid-disordered domains in model membranes, and how these approaches can aid in elucidating the properties of rafts in biological membranes. PMID- 16271404 TI - Effect of intraperitoneal CCK-8 on food intake and brain orexin-A after 48 h of fasting in the rat. AB - We investigated the interactions of the peripheral satiety peptide cholecystokinin and the brain orexin-A system in the control of food intake. The effect of an intraperitoneal (i.p.) injection of sulfated cholecystokinin octapeptide (in this article called CCK) (5 microg/kg, 4.4 nmol/kg) or of phosphate-buffered saline (PBS, vehicle control) on 48 h fasting-induced feeding and on orexin-A peptide content was analyzed in diverse brain regions innervated by orexin neurons and involved in the control of food intake. Administration of CCK after a 48 h fast reduced fasting-induced hyperphagia (P<0.05). I.p. CCK increased the orexin-A content in the posterior brainstem of 48 h fasted rats by 35% (P<0.05). Fed animals receiving CCK had 48% higher orexin-A levels in the posterior brainstem than fasted rats (P<0.05). In the lateral hypothalamus, fasting decreased orexin-A levels by 50% as compared to fed rats (P<0.05). In the septal nuclei, the combination of fasting and CCK administration reduced orexin-A contents compared to fed PBS and CCK animals by 13% and 17%, respectively (P<0.05). These results suggest a convergence of pathways activated by peripheral CCK and by fasting on the level of orexin-A released in the posterior brainstem and provide evidence for a novel interaction between peripheral satiety signaling and a brain orexigen in the control of food intake. PMID- 16271406 TI - Neutropenia associated with clopidogrel use in a patient with chronic renal failure who underwent percutaneous coronary and peripheral intervention. PMID- 16271407 TI - Facilitated recognition of an undeployed stent with StentBoost. AB - Underdeployment of stents is a major cause of restenosis. Standard cinegraphic visualisation has its shortcomings. We present a case in which we used a new technique, called StentBoost. This new technique enabled us to diagnose an undeployed stent as cause of a proximal restenosis problem, with direct major therapeutic consequences. PMID- 16271408 TI - The rapid evaluation of bacterial growth in blood cultures by selected ion flow tube-mass spectrometry (SIFT-MS) and comparison with the BacT/ALERT automated blood culture system. AB - We compared the performance of the BacT/ALERT automated blood culture system with real-time, quantitative volatile organic compound (VOC) detection by selected ion flow tube-mass spectrometry (SIFT-MS). Blood samples from healthy donors were artificially infected with 5 or 100 CFU of organisms commonly causing bacteremia. Positive results by SIFT-MS analysis of headspace gases were recorded for 53/60 (88.3%) bottles at 8h, and 58/60 (96.6%) bottles at 24 h. We conclude that SIFT MS is a sensitive method for the detection of microbial VOCs. Furthermore, profiles of the VOCs detected may allow simultaneous identification of infecting organisms. PMID- 16271409 TI - CT localization of axillary lymph nodes in relation to the humeral head: significance of arm position for radiation therapy planning. AB - Irradiation of the axillary nodes is often indicated in the clinical radiotherapy of patients with lymphoma and breast cancer. The relative location of the axillary nodes and the humeral head has historically been estimated based on lymphangiographic data. Since CT offers more precise definition, the axillary nodal region in relation to the humeral head was systematically studied in 61 patients undergoing CT simulation for breast conservation therapy. Differences in arm position (degree of abduction) significantly affected the location of the axillary nodal region in relation to the humeral head. With the arm abducted beyond 55 degrees, humeral head blocking would result in blocking of some of the axillary nodes. PMID- 16271410 TI - 3-D conformal radiotherapy of localized prostate cancer: a subgroup analysis of rectoscopic findings prior to radiotherapy and acute/late rectal side effects. AB - BACKGROUND AND PURPOSE: To identify endoscopic pathological findings prior to radiotherapy and a possible correlation with acute or chronic rectal side effects after three-dimensional conformal radiotherapy (3D-CRT) for prostate cancer. PATIENTS AND METHODS: Between 03/99 and 07/02, a total of 298 patients, who consented in a voluntary rectoscopy prior to radiotherapy were included into the analysis. Patients were treated with a total dose of either 70 or 74 Gy. Pathological rectoscopic findings like hemorrhoids, polyps or diverticula were documented. Acute and late rectal side effects were scored using the EORTC/RTOG score. RESULTS: The most frequent pathological endosopic findings were hemorrhoids (35%), polyps (24%) and diverticula (13%). Rectal toxicity was mostly low to moderate. Grade 0/1 cumulative acute and late rectal side effects were 82 and 84%, grade 2 were 18 and 17%, respectively. We could not identify any correlation between preexisting pathological findings and rectal side effects by statistical analysis. CONCLUSIONS: There is no evidence that prostate cancer patients presenting with endoscopic verified pathological findings in the rectal mucosa at diagnosis are at an increased risk to develop rectal side effects when treated with 3D-CRT of the prostatic region. PMID- 16271411 TI - Effect of gamma irradiation on microbial growth and sensory quality of fresh-cut lettuce. AB - Fresh-cut lettuce was irradiated and microbiological and sensory quality was examined during storage at 4 degrees C. Experimental results showed that the number of aerobic mesophilic bacteria on fresh-cut lettuce irradiated with 1.0 kGy was reduced by 2.35 logs and sensory quality was maintained best during storage for 8 days at 4 degrees C. It was indicated that Gompertz model could predict the bacterial number on fresh-cut lettuce if the initial level was known. PMID- 16271412 TI - Preservative effect of combined treatment with electrolyzed NaCl solutions and essential oil compounds on carp fillets during convectional air-drying. AB - The antimicrobial and antioxidant effects on carp fillet samples of treatments with alkaline electrolyzed NaCl solution EW (-) prior to treatment with acidic electrolyzed NaCl solution EW (+) and 1% solutions of the essential oils consisting of 0.5% carvacrol and 0.5% thymol (1% Cv+Ty) were tested. First carp fillet samples were treated with EW (-), then EW (+), followed by 1% (C+T), represented as [EW (-)/EW (+)/1% (Cv+Ty)] for 15 min, during drying at 45 degrees C. Samples were subsequently evaluated by microbiological, chemical and sensory analyses. Microbiological analyses indicated that the initial total microbial counts of samples treated with EW (-)/EW (+), 1% (Cv+Ty) or EW (-)/EW (+)/1% (Cv+Ty) were significantly (p< or =0.05) reduced, compared with the control sample. Treatment with EW (-)/EW (+)/1% (Cv+Ty) gave the strongest overall inhibition of microbial growth when compared to all of the other treatments. The volatile basic nitrogen (VBN) value of samples treated with EW (-)/EW (+)/1% (Cv+Ty) was kept at low level (18.46+/-0.45) until the end of drying period (5 days), compared with control samples (40.33+/-0.58). Treatment with EW (-)/EW (+)/1% (Cv+Ty) during drying significantly reduced the peroxide values (PV) and thiobarbituric acid values (TBA). Sensory evaluation indicated that there were significant differences (p< or =0.05) in the color, odor, taste, flavor and texture, on the end of the 5-day drying period between samples treated with EW ( )/EW (+)/1% (Cv+Ty), as compared to all of the other treatments. We conclude that treatment with EW (-)/EW (+)/1% (Cv+Ty) had stronger antimicrobial and antioxidant effects than all of the other treatments on carp fillets during drying, and could be a good alternative to artificial preservatives in food industry. PMID- 16271413 TI - Microbiological evaluation of bottled non-carbonated ("still") water from domestic brands in Greece. AB - The microbiological quality of 1,527 samples of bottled non-carbonated ('still') mineral water, purchased from retail outlets and derived from 10 manufacturing companies in Greece, was investigated during the period 1995-2003. Applying the membrane filter technique, the aliquots of water samples (250 ml) were analyzed for the presence and enumeration of total coliforms, Escherichia coli, Enterococcus spp. and Pseudomonas aeruginosa. Also, aerobic bacteria were counted as Heterotrophic Plate Count (HPC) ml(-1) at 22 and 37 degrees C. Positive samples for the parameters tested varied significantly among brands with an overall percentage of 13.95% bottled water samples noncompliant with the Greek water regulation. Microorganisms isolated from the samples tested were identified as species of Pseudomonas, Aeromonas, Pasteurella, Citrobacter, Flavobacterium, Providencia and Enterococcus. The most frequent isolated microorganism during the period of the study was P. aeruginosa. Generally, bacterial load of the samples tested ranged in low levels. The purpose of the current study was to evaluate the microbiological quality of the bottled water provided by domestic brands in the Greek market during the period 1995-2003. PMID- 16271414 TI - Oral administration of two probiotic strains, Lactobacillus gasseri CECT5714 and Lactobacillus coryniformis CECT5711, enhances the intestinal function of healthy adults. AB - Modifications in gastrointestinal parameters, intestinal colonization and tolerance are some of the main goals claimed for probiotics. However, although healthy people are the common target for these new functional food products, the number of clinical trials analysing the effects of probiotics in gastrointestinal parameters of healthy subjects is very scarce. A randomized, double blind, placebo-controlled human clinical trial involving 30 healthy adults was performed to investigate the effect of a fermented product containing two probiotic strains, Lactobacillus gasseri CECT5714 and Lactobacillus coryniformis CECT5711, on several blood and fecal parameters, most of them related to the host intestinal function. The volunteers were randomly distributed into two groups, one receiving a standard yogurt and the other a similar dairy fermented product in which the Lactobacillus delbreuckii subsp. bulgaricus yogurt strain had been replaced by a combination of the probiotic strains L. gasseri CECT5714 and L. coryniformis CECT5711. The volunteers that received the probiotic strains reported no adverse effects and the strains could be isolated from their feces at a relatively high level. In fact, the concentration of fecal lactic acid bacteria significantly increased in the probiotic group. Additionally, the oral administration of the probiotic strains led to an improvement of parameters such as the production of short chain fatty acids, the fecal moisture and the frequency and volume of the stools. As a result, the volunteers assigned to the probiotic group perceived a clear improvement in their intestinal habits. The study revealed that probiotics may exert a positive effect on healthy adults. PMID- 16271415 TI - The full-length clone of cucumber green mottle mosaic virus and its application as an expression system for Hepatitis B surface antigen. AB - A cucumber green mosaic mottle virus (CGMMV) full-length clone was developed for the expression of Hepatitis B surface antigen (HBsAg). The expression of the surface displayed HBsAg by the chimeric virus was confirmed through a double antibody sandwich ELISA. Assessment of the coat protein composition of the chimeric virus particles by SDS-PAGE analysis showed that 50% of the coat proteins were fused to the HBsAg. Biological activity of the expressed HBsAg was assessed through the stimulation of in vitro antibody production by cultured peripheral blood mononuclear cells (PBMC). PBMC that were cultured in the presence of the chimeric virus showed up to an approximately three-fold increase in the level of anti HBsAg immunoglobulin thus suggesting the possible use of this new chimeric virus as an effective Hepatitis B vaccine. PMID- 16271417 TI - Embryonic stem cells: understanding their history, cell biology and signalling. AB - Embryonic stem cells offer enormous potential as a source of a variety of differentiated cells for cell therapy, drug discovery and toxicology screening. With the creation of human embryonic stem cell lines we now have a resource with the potential to differentiate into every tissue of the body. To fully harness this resource it is necessary to understand their biology. Here we give a background to their history, describe interesting elements of their cell biology and introduce the underlying signalling mechanisms that control their ability to self-renew and differentiate. PMID- 16271416 TI - Deoxycholic acid-conjugated chitosan oligosaccharide nanoparticles for efficient gene carrier. AB - To develop chitosan based efficient gene carriers, highly purified chitosan oligosaccharides (COSs) were chemically modified with deoxycholic acid (DOCA). Owing to the amphiphilic characters, the DOCA-conjugated COSs (COSDs) formed self aggregated nanoparticles in aqueous milieu. The physicochemical characterization revealed that the particle size of the nanoparticles was in the range of 200 approximately 240 nm and the critical aggregation concentration (cacs) was 0.012 approximately 0.046 g/L, depending on the degree of substitution (DS). As efficient gene carriers, the COSD nanoparticles showed superior gene condensation and protection of condensed gene from endonuclease attack than unmodified COSs. Furthermore, COSDs showed great potential for gene carrier with the high level of gene transfection efficiencies, even in the presence of serum. Considered with the negligible cytotoxic effects, DOCA-modified chitosan oligosaccharides can be considered as potential candidates for efficient non-viral gene carriers. PMID- 16271418 TI - Molecularly imprinted polymers for the enantioseparation of chiral drugs. AB - Molecularly imprinted polymers have been applied for many years in chiral separations and increasingly have been applied to drugs. Separation speed and efficiency in methods such as HPLC and CEC are increasingly competitive with alternative established CSPs. The literature on separation of drug enantiomers using MIPs via HPLC, TLC, SFC and CEC are reviewed. Issues for method development, particularly the monomer-template interactions during the imprinting process, and the mechanism of solute retention on MIP CSPs in chromatography using normal phase or reverse-phase mobile phases are considered. PMID- 16271419 TI - The amyloid pathology progresses in a neurotransmitter-specific manner. AB - Past studies using transgenic models of early-staged amyloid pathology, have suggested that the amyloid pathology progresses in a neurotransmitter-specific manner where cholinergic terminals appear most vulnerable, followed by glutamatergic terminals and finally by somewhat more resistant GABAergic terminals. To determine whether this neurotransmitter-specific progression persists at later pathological stages, presynaptic bouton densities, and the areas of occupation and localization of plaque adjacent dystrophic neurites were quantified in 18-month-old APP(K670N, M671L)+PS1(M146L) doubly transgenic mice. Quantification revealed that transgenic animals had significantly lower cholinergic, glutamatergic and GABAergic presynaptic bouton densities. Cholinergic and glutamatergic dystrophic neurites appear to be heavily influenced by fibrillar Abeta as both types displayed a decreasing area of occupation with respect to increasing plaque size. Furthermore, cholinergic dystrophic neurites reside in closer proximity to plaques than glutamatergic dystrophic neurites, while GABAergic dystrophic neurites were minimal regardless of plaque size. To investigate whether similarities exist in the human AD pathology, a monoclonal antibody (McKA1) against the human vesicular glutamate transporter 1 (VGluT1) was developed. Subsequent staining in AD brain tissue revealed the novel presence of glutamatergic dystrophic neurites, to our knowledge the first evidence of a structural glutamatergic deficit in the AD pathology. PMID- 16271420 TI - Early neuropathology of somatostatin/NPY GABAergic cells in the hippocampus of a PS1xAPP transgenic model of Alzheimer's disease. AB - At advanced stages, Alzheimer's disease (AD) is characterized by an extensive neuronal loss. However, the early neurodegenerative deficiencies have not been yet identified. Here we report an extensive, selective and early neurodegeneration of the dendritic inhibitory interneurons (oriens-lacunosum moleculare, O-LM, and hilar perforant path-associated, HIPP, cells) in the hippocampus of a transgenic PS1xAPP AD model. At 6 months of age, from 22 different pre- and postsynaptic mRNA markers tested (including GABAergic, glutamatergic and cholinergic markers), only the expression of somatostatin (SOM) and NPY neuropeptides (O-LM and HIPP markers) displayed a significant decrease. Stereological cell counting demonstrated a profound diminution (50-60%) of SOM immunopositive neurons, preceding the pyramidal cell loss in this AD model. SOM population co-expressing NPY was the most damaged cell subset. Furthermore, a linear correlation between SOM and/or NPY deficiency and Abeta content was also observed. Though the molecular mechanism of SOM neuronal loss remains to be determined, these findings might represent an early hippocampal neuropathology. Therefore, SOM and NPY neuropeptides could constitute important biomarkers to assess the efficacy of potential early AD treatments. PMID- 16271421 TI - Synthesis, characterization and biological studies of Co(II), Ni(II), Cu(II) and Zn(II) complexes with bidentate Schiff bases derived by heterocyclic ketone. AB - A series of metal complexes of Co(II), Ni(II), Cu(II) and Zn(II) have been synthesized with newly prepared biologically active ligands. These ligands were prepared by the condensation of 4-amino-5-mercapto-3-methyl-s-triazole (AMMT), 4 Amino-3-ethyl-5-mercapto-s-triazole (AEMT) with 2-acetylpyridine. The structure of the complexes have been proposed by elemental analyses, spectroscopic data i.e. IR, 1H NMR, electronic and magnetic measurements. Thermal studies of the complexes are also reported. Antibacterial activities of 10 complexes have been studied in vitro. Heterocyclic bidentate Schiff bases were associated with substantially higher antibacterial activities than some commercial antibiotics. PMID- 16271422 TI - Growth characteristics of canine pathogenic viruses in MDCK cells cultured in RPMI 1640 medium without animal protein. AB - Madin Darby canine kidney (MDCK) cells were adapted to serum-free RPMI 1640 medium and used for cultivation of canine viruses. RPMI 1640 medium was supplemented with a soybean peptone, L-glutamine and antibiotics, so that the protein concentration was less than 5 microg/ml (RPMI/SP medium). The resulting adapted MDCK-SP cells showed steady growth after the twenty-eighth passage in RPMI/SP medium (MDCK-SP cell culture). Canine distemper virus, canine parvovirus, canine adenoviruses and canine parainfluenza virus, which are the principal components of canine combined virus vaccines, grew in the MDCK-SP cell culture as efficiently as the parental MDCK cells cultured in the conventional Eagle's MEM containing fetal bovine serum. Consequently, the use of MDCK-SP cell culture can make current canine vaccine products much safer, of higher quality and at lower cost. PMID- 16271423 TI - Anticipating crisis: towards a pandemic flu vaccination strategy through alignment of public health and industrial policy. AB - Flu pandemics (worldwide epidemics) have occurred at irregular and unpredictable intervals, and have been associated with substantial morbidity, mortality and economic cost. In response to the emerging potential for a new pandemic to occur, national and international preparedness plans are being drawn up specifying the use of antivirals and vaccines. A number of challenges to pandemic vaccine development, large-scale production and the timing of distribution have also been highlighted. This article reviews the rationale and consequential policy for aligned public- and private sector planning in the present inter-pandemic period despite the prevalent risks and uncertainties. We propose a model for product development of pandemic flu vaccine based on public-private partnership, including push and pull incentive mechanisms for stimulating work in this therapeutic area. In addition, we argue that innovative vaccination strategies, together with special vaccine formulations which may offer cross-protection against multiple flu pandemic strains might avert the worse effects of an influenza infection. PMID- 16271424 TI - Long-term clinical trial safety experience with the inactivated split influenza vaccine, Vaxigrip. AB - Safety data on the inactivated split influenza vaccine, Vaxigrip, were compiled and analysed from 28 clinical trials (total: 4599 subjects aged 6 months to 99 years) to provide a robust estimate of the reactogenicity profile. The most frequent solicited reactions were non-severe injection site pain and erythema in children, adults, and elderly. Mild or moderate fever was the most frequent reaction in 6-36 months olds; few systemic reactions were reported in older groups. Reactogenicity was comparable in healthy and high-risk children. The long term experience with the world's most widely used influenza vaccine, Vaxigrip, confirms its excellent tolerability, and supports its continued use in clinical practice worldwide. PMID- 16271425 TI - Functional recovery of biofilm bacterial communities after copper exposure. AB - Potential of bacterial communities in biofilms to recover after copper exposure was investigated. Biofilms grown outdoor in shallow water on glass dishes were exposed in the laboratory to 0.6, 2.1, 6.8 micromol/l copper amended surface water and a reference and subsequently to un-amended surface water. Transitions of bacterial communities were characterised with denaturing gradient gel electrophoresis (DGGE) and community-level physiological profiles (CLPP). Exposure to 6.8 micromol/l copper provoked distinct changes in DGGE profiles of bacterial consortia, which did not reverse upon copper depuration. Exposure to 2.1 and 6.8 micromol/l copper was found to induce marked changes in CLPP of bacterial communities that proved to be reversible during copper depuration. Furthermore, copper exposure induced the development of copper-tolerance, which was partially lost during depuration. It is concluded that bacterial communities exposed to copper contaminated water for a period of 26 days are capable to restore their metabolic attributes after introduction of unpolluted water in aquaria for 28 days. PMID- 16271426 TI - Woody biomass phytoremediation of contaminated brownfield land. AB - Economic and environmental regeneration of post-industrial landscapes frequently involves some element of re-afforestation or tree planting. We report field trials that evaluate whether woody biomass production is compatible with managing residual trace element contamination in brownfield soils. Large-scale mapping of contamination showed a heterogenous dispersion of metals and arsenic, and highly localised within-site hotspots. Yields of Salix, Populus and Alnus were economically viable, showing that short-rotation coppice has a potentially valuable role in community forestry. Mass balance modelling demonstrated that phytoextraction potentially could reduce contamination hotspots of more mobile elements (Cd and Zn) within a 25-30-year life cycle of the crops. Cd and Zn in stems and foliage of Salix were 4-13 times higher than EDTA-extractable soil concentrations. Lability of other trace elements (As, Pb, Cu, Ni) was not increased 3 years after planting the coppice; woody biomass may provide an effective reduction of exposure (phyto-stabilization) to these less mobile contaminants. PMID- 16271427 TI - Ecological impact assessment of metallurgic effluents using in situ biomarker assays. AB - An ecological impact study was performed based on in situ biomarker assays with the waterflea Daphnia magna. The effects of metallurgic effluents on the energy metabolism, anti-oxidative metabolism and DNA damage were assessed in caged daphnids during a 4-week study. In situ survival and reproduction studies demonstrated a clear impact on these parameters in organisms exposed in the most polluted areas. At the downstream--sublethal--zone the organisms were disturbed within their tolerance limits, resulting in alterations of their energy metabolism. These data suggest an acclimation hypothesis, which was tested through the analysis of the energy metabolism of resident species: isopods and amphipods. These organisms had shifted to a decrease in their overall energy metabolism compared to the upstream region. This change in some biochemical processes suggests a selective advantage to cope with the prevailing environmental conditions. In addition, we found clear genotoxic effects caused by the industrial discharges that might correlate with a reduction in (long-term) survival. PMID- 16271428 TI - Characterizing the risk assessment of heavy metals and sampling uncertainty analysis in paddy field by geostatistics and GIS. AB - For many practical problems in environmental management, information about soil heavy metals, relative to threshold values that may be of practical importance is needed at unsampled sites. The Hangzhou-Jiaxing-Huzhou (HJH) Plain has always been one of the most important rice production areas in Zhejiang province, China, and the soil heavy metal concentration is directly related to the crop quality and ultimately the health of people. Four hundred and fifty soil samples were selected in topsoil in HJH Plain to characterize the spatial variability of Cu, Zn, Pb, Cr and Cd. Ordinary kriging and lognormal kriging were carried out to map the spatial patterns of heavy metals and disjunctive kriging was used to quantify the probability of heavy metal concentrations higher than their guide value. Cokriging method was used to minimize the sampling density for Cu, Zn and Cr. The results of this study could give insight into risk assessment of environmental pollution and decision-making for agriculture. PMID- 16271429 TI - Responses of antioxidant systems and LPO level to benzo(a)pyrene and benzo(k)fluoranthene in the haemolymph of the scallop Chlamys ferrari. AB - The effects of benzo(a)pyrene (BaP), benzo(k)fluoranthene (BkF) and their mixture on antioxidant enzyme activities and lipid peroxidation (LPO) levels of haemolymph of scallop (Chlamys ferrari) were studied. The superoxide dismutase (SOD) activities of 0.5 microg/L and 1.0 microg/L were significantly higher than controls (P<0.05), while it increased at beginning and then dropped (lower than controls) in the end at 10.0 microg/L and 50.0 microg/L PAHs groups. The catalase (CAT) activities were very little during the whole experimental time. The glutathione peroxidase (GPx) activities in each PAHs group all increased significantly (P<0.05). LPO levels all increased significantly (P<0.05) with time at each PAHs group except for the 0.5 microg/L group of less than hour 12. The toxicity of PAHs in a descending order was BaP>BkF>mixture of BaP and BkF. The changes in antioxidant enzyme activities and LPO level in haemolymph could reflect the detoxification functions and damage levels of whole organism. PMID- 16271430 TI - MEAD: an interdisciplinary study of the marine effects of atmospheric deposition in the Kattegat. AB - This paper summarises the results of the EU funded MEAD project, an interdisciplinary study of the effects of atmospheric nitrogen deposition on the Kattegat Sea between Denmark and Sweden. The study considers emissions of reactive nitrogen gases, their transport, transformations, deposition and effects on algal growth together with management options to reduce these effects. We conclude that atmospheric deposition is an important source of fixed nitrogen to the region particularly in summer, when nitrogen is the limiting nutrient for phytoplankton growth, and contributes to the overall eutrophication pressures in this region. However, we also conclude that it is unlikely that atmospheric deposition can, on its own, induce algal blooms in this region. A reduction of atmospheric nitrogen loads to this region will require strategies to reduce emissions of ammonia from local agriculture and Europe wide reductions in nitrous oxide emissions. PMID- 16271431 TI - Effects of oxygen on the release and distribution of phosphorus in the sediments under the light condition. AB - Effects of oxygen on the release and distribution of phosphorus (P) in the sediments in the presence of light were investigated, using sediment cores and overlying water from Lake Taihu, in China. The results show that P can be released from sediments to the overlying water in both anoxic and aerobic conditions. But more P was released in the anoxic condition. The transformation of P between various fractions in the sediments was observed during the release experiments. Concentrations of Ca-bound P and organic P in the sediments decreased in both conditions, but Fe/Al-bound P increased in the aerobic condition. The decrease of total P and P fractions in the sediments is consistent with the accumulative increase in quantity (AIQ) of total P (TP) in the overlying water, but is contrary with the AIQ of dissolved inorganic P. This is due to the uptake of algae by the dissolved inorganic P. Total nitrogen in the sediments in the anoxic condition was lower than that in the aerobic condition, and pH in the overlying water increased in the anoxic condition. PMID- 16271432 TI - TDI can induce respiratory allergy with Th2-dominated response in mice. AB - Toluene diisocyanate (TDI), a highly reactive industrial chemical is one of the leading causes of occupation-related asthma in industrialized countries. The pathophysiology of TDI-induced asthma, however, remains poorly understood, in part due to a lack of appropriate animal models. In this study, four models of TDI-sensitised mice were investigated. In model number 1, the mice were sensitised for 4 h/day on four consecutive days to 3 ppm inhaled TDI and challenged twice for 4 h each time with 0.3 ppm inhaled TDI. In model number 2, the sensitising condition was similar to that of model 1, but the challenge conditions involved an initial inhalation of 2 ppmTDI for 4h and then tracheal instillation with 50 microg/mouse albumin-TDI. In model number 3, the mice were sensitised first to 25% TDI (sc) and then three times for 4 h each time to 1 ppm inhaled TDI and challenged twice for 4h each time with 0.1 ppm inhalated TDI. In model number 4, the mice were first sensitised to 1% TDI by skin application and then with 0.2% TDI by tracheal instillation and challenged tree times by tracheal instillation of 0.1% TDI. In model number 4, skin application followed by tracheal instillations of TDI led to local and systemic Th2-dominated immune responses that were characterized: (1) in the lung-associated lymph nodes by a decrease in Th1 cytokine (IFN-gamma) production associated with an increase in Th2 cytokine (IL-4, IL-5, IL-3) production; (2) in the lungs by an allergic inflammation throughout the conducting airways: goblet cell proliferation and eosinophil influx and; (3) in the serums by increased total and specific IgE levels, 17.5- and 3.5-fold higher than that of the controls, respectively. The conditions used for sensitisation in the other models, i.e. inhalation or subcutaneous administration plus inhalation, failed to induce a strong Th2 response like that observed in model number 4. The findings indicate that TDI can induce a Th2-dominated response in mice when administered by topical application plus tracheal instillation for sensitisation and by intra-tracheal instillation for challenge (model number 4). This mouse Th2 model of TDI-induced airway allergy can, in several aspects, mimic occupational TDI asthma in humans and may prove to be useful in determining the mechanistic basis behind this disease. PMID- 16271433 TI - Regeneration of descending axon tracts after spinal cord injury. AB - Axons within the adult mammalian central nervous system do not regenerate spontaneously after injury. Upon injury, the balance between growth promoting and growth inhibitory factors in the central nervous system dramatically changes resulting in the absence of regeneration. Axonal responses to injury vary considerably. In central nervous system regeneration studies, the spinal cord has received a lot of attention because of its relatively easy accessibility and its clinical relevance. The present review discusses the axon-tract-specific requirements for regeneration in the rat. This knowledge is very important for the development and optimalization of therapies to repair the injured spinal cord. PMID- 16271434 TI - Relationship between homocysteine and neopterin concentrations in patients with gynecological cancer. AB - Elevated concentrations of vascular risk factor homocysteine have been described in patients with malignant diseases, and homocysteine was supposed to be useful as tumor marker. Likewise, elevated concentrations of Th1-type immune activation marker neopterin are frequently observed in patients suffering from cancer and serve as prognostic marker for the survival of patients. In this study, the relationship between homocysteine and neopterin concentrations was examined in 18 patients with gynecological cancer. Concentrations of homocysteine and cysteine were measured by HPLC in sera of patients, folic acid and vitamin B(12) levels were determined by radioimmunoassay, and neopterin concentrations were measured by ELISA. Median homocysteine concentration was 11.2 microM (interquartile range: 9.9-13.2 microM), 3 patients had levels higher than 15 microM, the upper limit of the normal range. Neopterin concentrations were increased in 13 patients (median: 11.6; 7.7-24.9 nM), cysteine (median: 234; 216-255 microM), folate (median: 7.8; 6.2-11.7 ng/ml) and vitamin B(12) (median: 352; 258-570 pg/ml) concentrations were all within reference ranges although rather at the lower side. Higher homocysteine concentrations correlated inversely with low folate concentrations (r(s)=-0.605; P<0.01) and tended to be higher in patients with higher cysteine levels (r(s)=0.457; P<0.06; ). No correlation was found between homocysteine and immune activation marker neopterin, although the three patients with elevated homocysteine concentrations tended to have higher neopterin levels as well (P<0.07). In conclusion, only a few patients with gynecological cancer present with elevated homocysteine concentrations and hyperhomocysteinemia seems only weakly related to immune activation phenomena, tumor cell proliferation probably is more important for the increase of homocysteine. PMID- 16271435 TI - A new targeting approach for breast cancer gene therapy using the heparanase promoter. AB - Gene therapy with adenoviral (Ad) vectors is a promising new approach in the treatment of cancer. Strategies to restrict adenoviral-mediated transgene expression are important to avoid gene transfer into normal cells. Heparanase (HPR) is overexpressed in breast cancer but downregulated in differentiated normal tissue. Expression of the HPR gene was evaluated in breast cancer cells. Biodistribution and liver tropism was evaluated in a mouse model. HPR is highly expressed in breast cancer tissue. The HPR promoter retained its fidelity in an adenovirus context and was activated in breast cancer cells but showed low activity in normal breast cells and the murine liver. We conclude that the HPR pathway is a promising target for the development of breast cancer directed gene therapy strategies. PMID- 16271436 TI - Differentiation and growth inhibition mediated via the RXR:PPARgamma heterodimer in colon cancer. AB - This study evaluated the anti-tumor efficacy of combining the RXR agonist, bexarotene, with the PPARgamma agonist, rosiglitazone, in colon cancer. Moser, a human colon cancer cell line, was treated with bexarotene and rosiglitazone alone or in combination and the effect on growth and differentiation were examined. The data demonstrated that the bexarotene/rosiglitazone combination produced greater efficacy in growth inhibition than either single agent. Furthermore, combination treatment acted cooperatively to decrease COX-2 expression and PGE2 synthesis while increasing expression of the differentiation marker, CEA. These findings were confirmed in vivo in a Moser xenograft tumor model. Collectively, our data suggest a potential role for utilizing a combination regimen of a RXR and PPARgamma agonist in the treatment of colon cancer. PMID- 16271437 TI - Protection against UV-light-induced skin carcinogenesis in SKH-1 high-risk mice by sulforaphane-containing broccoli sprout extracts. AB - Aerobic life, UV solar radiation, genetic susceptibility, and immune status contribute collectively to the development of human skin cancers. In addition to direct DNA damage, UV radiation promotes the generation of reactive oxygen intermediates that can cause oxidative damage and inflammation, and ultimately lead to tumor formation. Treatment of murine and human keratinocytes with the isothiocyanate sulforaphane elevated phase 2 enzymes and glutathione and protected against oxidant toxicity. Topical application of sulforaphane containing broccoli sprouts extracts induced the phase 2 response in mouse skin in vivo. Sulforaphane inhibited cytokine-dependent (gamma-interferon or lipopolysaccharide) induction of iNOS in RAW 264.7 macrophages. The UV-radiation induced skin carcinogenesis in "initiated high-risk mice" was substantially inhibited by broccoli sprout extracts containing sulforaphane. After completion of the UV irradiation schedule (30 mJ/cm(2)/session twice a week for 20 weeks), groups of approximately 30 mice were treated topically on their backs (5 days a week for 11 weeks) with broccoli sprout extract containing either the equivalent to 0.3 micromol (low dose) or 1.0 micromol (high dose) sulforaphane, respectively. At this time point, the tumor incidence had reached 100% in the control mice. Tumor burden, incidence, and multiplicity were reduced by 50% in the animals that received the high dose of protector. Tumor incidence and multiplicity did not differ between the low dose-treated and the control groups, but the low dose treatment resulted in a substantial reduction of the overall tumor burden. Thus, topical application of sulforaphane-containing broccoli sprout extracts is a promising strategy for protecting against skin tumor formation after exposure to UV radiation. PMID- 16271439 TI - Rothmund-Thomson syndrome and RECQL4 defect: splitting and lumping. AB - Rothmund-Thomson Syndrome (RTS) is a rare autosomal recessive genodermatosis with a heterogeneous clinical profile. Mutations in RECQL4, encoding a RecQ DNA helicase, are present in a large fraction, but not all clinically diagnosed patients, allowing to classify RTS among the RecQ helicase chromosomal instability defects including Bloom's and Werner's syndromes. Results of RECQL4 test coupled to the variable clinical presentation favored the splitting of RTS clinical phenotype into nosological entities under distinct genetic control. In parallel, lumping of the RECQL4 gene to two other diseases, RAPADILINO and Baller Gerold has paved the way to unravel through allelic heterogeneity complex genotype-phenotype correlations. Recql4 knockout mice provided crucial insights into the comprehension of the functional role of RECQL4 helicase, which have been corroborated by the initial biochemical characterization of RECQL4 protein and its acting pathway and by studies on RECQL4 homologs in yeast and Xenopus. A role for RECQL4 in initiation of DNA replication and in sister chromatid cohesion has been proposed, which currently fits the pieces of evidence achieved by different approaches. Further work is needed to define the specific and shared functions of RECQL4 in relation to other RecQ helicases and to connect RECQL4 diseases to other genomic instability syndromes with birth defects and cancer predisposition. PMID- 16271440 TI - Cell hydration as the primary factor in carcinogenesis: A unifying concept. AB - The paper discusses the unifying concept that cell hydration is the primary factor in the mechanism of carcinogenesis. The concept includes the following hypotheses: (1) Increased cell hydration causes cancer not only by promoting cell division and oncogene expression, but also by inactivating genes inducing cell differentiation, and by preventing apoptosis. Conversely, factors that reduce cell hydration prevent cancer by inhibiting cell division and oncogene expression, while activating genes inducing cell differentiation, and by promoting apoptosis. The unique ability of cell hydration to have these opposite effects on cell behavior and gene expression can account for its postulated role as the primary factor in both the promotion and prevention of cancer. (2) A progressive increase in cell hydration, induced by successive mutations and/or epigenetic changes, is the basic mechanism of multi-step carcinogenesis, the degree of malignancy increasing with the degree of cell hydration. (3) The increased hydration of cancer cells accelerates their respiration rate, thereby enhancing their ability to compete for nutrients with their normal counterparts. This effect may play a major role in promoting tumor growth and in the postulated mechanism of multi-step carcinogenesis. (4) Increased cell hydration is also proposed as an alternative or additional explanation of the carcinogenetic effect of inflammatory agents and of hormones. A survey of the literature provides evidence consistent with these hypotheses, but suggestions are included for further investigations to test their validity and their implications. From a clinical perspective, the abnormally high water content of cancer cells permits the use of microwave technology for tumor detection and treatment. Also of considerable therapeutic significance is the increased sensitivity if cancer cells to desiccation, postulated to result from genetic changes induced by increased hydration. This may well be the achilles heel of cancer, and recent investigations indicate that it may be exploited very effectively in the treatment of the disease. In conclusion, I suggest that the need for studies on the molecular biology of cancer to be supplemented by more information on environmental effects on gene expression and on the biochemical and physiological factors that mediate genetic effects at the cellular level. This approach might also be used to assess the validity of the postulated role of cell hydration as a factor of particular significance. PMID- 16271438 TI - Cytotoxicity of liposomal alpha-tocopheryl succinate towards hamster cheek pouch carcinoma (HCPC-1) cells in culture. AB - There is compelling evidence for the cancer chemopreventive effects of vitamin E and related compounds. Of all the vitamin E derivatives that have been investigated to date, vitamin E acid succinate is the most effective anti-cancer agent. This report describes the preparation and testing of liposomal formulation of mono alpha-tocopheryl ester of succinic acid (alpha-TOS) for cytotoxicity against hamster cheek pouch carcinoma cell line (HCPC-1). Small unilamellar vesicles (SUV) of phosphatidylcholine incorporating 70 microM alpha-TOS were superior to alpha-TOS alone or SUV without incorporated alpha-TOS, as inducers of apoptosis in HCPC-1 cells. Liposomal alpha-TOS perturbed the lipid structure in cells, promoted apoptosis, and decreased cell viability. The mechanism of action of alpha-TOS appears to involve membrane damage and induction of ceramide mediated apoptosis. PMID- 16271441 TI - Clinical outcome of hypofractionated conventional conformation radiotherapy for patients with single and no more than three metastatic brain tumors, with noninvasive fixation of the skull without whole brain irradiation. AB - PURPOSE: To evaluate the efficacy and toxicity of hypofractionated conventional conformation radiotherapy (HCCRT) with noninvasive fixation of the skull on patients with single or several brain metastases. METHODS AND MATERIALS: The subjects were 44 patients who had three or fewer brain metastases (26 solitary, 18 multiple). Treatment was conducted on 65 metastases by rotational conformal beam or multiple fixed coplanar beams with a standard linear accelerator. The planning target volume consisted of the tumor and a 1-cm safety margin. The median isocentric dose was 24 Gy (range, 18-30 Gy) in 3-5 fractions. Whole-brain irradiation was not applied as an initial treatment. RESULTS: Actuarial local tumor control rates at 6 months and 1 year were 78.4% and 71.9%, respectively. In field recurrence was noted in 10 of 65 tumors, and repeat HCCRT was applied in 5 tumors. Actuarial overall survival rates at 1 year, 2 years, and the median survival time were 50.8%, 24.1%, and 5.8 months, respectively. The patients with an active primary cancer and poor performance status had a poorer prognosis than those without those factors. Actuarial freedom from second brain metastases rates at 6 months, 1 year, and 2 years were 86.6%, 69.0%, and 40.9%, respectively. Second brain metastases were observed in 9 of 44 patients. Lung adenocarcinomas had a higher risk of second brain metastasis than others. Treatment-related severe early or late complications were not observed in this series. CONCLUSIONS: Hypofractionated conventional conformation radiotherapy achieved sufficient tumor control and survival. The results suggest that HCCRT would be one of the alternatives for patients with either solitary or several brain metastases. PMID- 16271442 TI - Correlation between MR imaging-derived nasopharyngeal carcinoma tumor volume and TNM system. AB - PURPOSE: To measure nasopharyngeal carcinoma tumor volume based on magnetic resonance images using a validated semiautomated measurement methodology and correlate tumor volume with TNM T classification. METHODS AND MATERIALS: The study population consisted of 206 consecutive nasopharyngeal carcinoma patients who had magnetic resonance imaging staging scans. Tumor volume was measured using a semisupervised knowledge-based fuzzy clustering algorithm. Patients were divided into 4 groups according to TNM T classification. The difference in tumor volumes among the various TNM T-classification groups was examined. RESULTS: The mean tumor volume in each T-classification group is as follows: T1, 8.6 mL +/- 5.0 (standard deviation [SD]); T2, 18.1 mL +/- 8.1 (SD); T3, 25.8 mL +/- 14.1 (SD); and T4, 36.2 mL +/- 18.9 (SD). The mean tumor volume increased significantly with advancing T classification (p < 0.0001). Tumor volume in a more advanced T group was significantly larger than that in an adjacent early T group (p < 0.01). CONCLUSION: Validated magnetic resonance imaging-based tumor volume shows positive correlation between tumor volume and advancing T classification groups. It may be possible to incorporate tumor volume as an additional prognostic factor into the existing TNM system. PMID- 16271443 TI - Antihyperglycemic and antilipidperoxidative effects of Pongamia pinnata (Linn.) Pierre flowers in alloxan induced diabetic rats. AB - Our aim was to evaluate the antihyperglycemic and antilipid peroxidative effect of ethanolic extract of Pongamia pinnata (Linn.) Pierre (Leguminosae) flowers (PpEt) in normal rats and alloxan induced diabetic rats. Hyperglycemia, elevated lipid peroxidation [thiobarbituric acid reactive substances (TBARS)] and disturbed nonenzymatic [Vitamin E, Vitamin C and glutathione] and enzymatic antioxidants status were noticed in alloxan induced diabetic rats. The oral administration of ethanolic extract of Pongamia pinnata flowers (300 mg/kg bw) showed significant antihyperglycemic, and antilipidperoxidative effects and enhancement in antioxidants defense system in alloxan induced diabetic rats. However, no significant characteristic changes were noticed in blood glucose level as well as in lipid peroxidation and antioxidant status in normal rats treated with "PpEt" alone. We have also observed that the "PpEt" considerably reduced the blood glucose concentration in a similar extent to that of the reference drug glibenclamide (600 microg/kg bw) in alloxan induced diabetic rats. Our results thus suggested that the "PpEt" could be used as a safe alternative antihyperglycemic drug for diabetic patients. PMID- 16271444 TI - Age-related changes in circadian rhythm of serotonin synthesis in ring doves: effects of increased tryptophan ingestion. AB - Alterations in the function of the hypothalamic suprachiasmatic nucleus (SCN) with age have been reported. As serotonin is an important regulator of the circadian clock located in SCN, this work studied the changes produced in the synthesis of serotonin with age using the accumulation of 5-HTP after decarboxylase inhibition as a measure of serotonin synthesis in the brain in vivo, in young and old ring doves at the onset of lights-on and lights-off. A diurnal cycle in tryptophan hydroxylation was observed in young animals, with an increased daylight synthesis and metabolism of 5-HT in hippocampus, neostriatum and hypothalamus. A single dose of melatonin (1 mg/kg, i.p., 1 h) at lighttime produced an inhibitory effect on the synthesis of 5-HT. In contrast, differences in 5-HT synthesis and metabolism between day and night disappeared in old animals indicating an absence of a circadian rhythm in 5-HT synthesis and metabolism. The administration of L-tryptophan (240 mg/kg, i.p.) strongly increased the 5-HT synthesis in young animals only during lights-off time while it increased in old ones irrespective of the administration time. These results suggest that the supplemental administration of tryptophan might aid to improve the descent in 5 HT that normally occurs, as animals get old. PMID- 16271445 TI - [Cerebellum abscess: first demonstration of undiagnosed infective endocarditis in an adult with corrected transposition arteries]. AB - Neurological complications occur in approximately 30% of all patients with infective endocarditis and represent a major factor associated with an increased mortality rate. Third of these complications is represented by cerebral embolism, followed by mycotic aneurysm, meningitis or meningoencephalitis. Brain abscesses are rare; their localization to the cerebellum is exceptional. A case of cerebellum abscess occurring in a 34-year-old patient with non-operated corrected transposition of the great arteries is reported. Occurrence of this abscess was the first demonstration of undiagnosed infective endocarditis. PMID- 16271446 TI - Pharmacogenetics of irinotecan metabolism and transport: an update. AB - The anticancer agent irinotecan (CPT-11) is converted to SN-38, which is approximately 100 to 1,000-fold more cytotoxic than the parent drug. The pharmacokinetics of irinotecan are extremely complex and have been the subject of intensive investigation in recent years. Irinotecan is subject to extensive metabolism by various polymorphic enzymes, including CES2 to form SN-38, members of the UGT1A subfamily, and CYP3A4 and CYP3A5, which form several pharmacologically inactive oxidation products. Elimination of irinotecan is also dependent on drug-transporting proteins, notably ABCB1 (P-glycoprotein), ABCC2 (cMOAT) and ABCG2 (BCRP), present on the bile canalicular membrane. The various processes mediating drug elimination, either through metabolic breakdown or excretion, likely impact substantially on interindividual variability in drug handling. This report provides an update on current strategies to individualize irinotecan chemotherapy based on each patient's genetic constitution, which may ultimately lead to more selective use of this agent. PMID- 16271447 TI - Developmental toxicity of meglumine antimoniate and transplacental transfer of antimony in the rat. AB - Meglumine antimoniate (MA), a pentavalent antimonial (SbV) drug, has been used for the treatment of leishmaniases for over half a century but there is almost no data on its safety and kinetics during pregnancy. This study was undertaken to investigate the developmental toxicity of MA as well as the transplacental transfer of antimony (Sb) in rats. Wistar rats (approximately 20 per group) were treated subcutaneously (s.c.) with MA (0, 75, 150, 300 mg SbV/(kg BW day)) during pregnancy (days 1-20). An untreated control group was evaluated as well. Caesarean sections were performed on day 21 and implantations, living and dead fetuses, and resorptions were recorded. Fetuses were weighed and fixed in Bouin's solution and one-third of each litter was examined for soft-tissue anomalies. The remaining fetuses were cleared and stained with Alizarin red S for skeleton evaluation. No adverse effect of MA on the mothers was noted at any dose level. No embryotoxicity was observed at the lowest dose but, at the highest dose, MA increased embryolethality, reduced fetal weight and augmented the occurrence of some soft-tissue and skeleton variations. Therefore, the no-observed-adverse effect level for MA-induced embryotoxicity was 75 mg SbV/(kg BW day). In a separate group of rats treated with 300 mg SbV/(kg BW day) s.c. during whole pregnancy, blood (0.2 mL) was taken from the tail vein 1, 2, 4, 6, 12 and 24 h after treatment on day 1 and thereafter every other day immediately before drug injection. Blood was also taken from fetuses removed on day 21, 24h after the last injection of MA. Blood levels of Sb were determined by ICP-MS and results showed that most of administered Sb was eliminated rapidly (in less than 6 h), but nadir blood concentrations increased gradually during treatment from 1 to 2 microg/g, 24h after the first dose, up to approximately 38 microg/g after the 20th dose. Levels of Sb in fetal blood were as high as 10-15 microg/g, i.e. approximately 30% their mothers' nadir levels near term. These findings indicated that repeated administration of MA during gestation led to an accumulation of Sb in mothers and fetuses. PMID- 16271448 TI - Detection of Shiga toxin genes stx1, stx2, and the +93 uidA mutation of E. coli O157:H7/H-using SYBR Green I in a real-time multiplex PCR. AB - Enterohemorrhagic Escherichia coli (EHEC) is a major foodborne pathogen capable of causing diarrhea and vomiting, but more serious complications such as hemorrhagic colitis and hemolytic-uremic syndrome (HUS) can result. A real-time PCR method to detect the presence of Shiga toxin producing E. coli (STEC) and E. coli O157:H7 was investigated using SYBR Green I (SG). Primers were designed to target the Shiga toxin genes (stx1 and stx2) and a highly conserved base substitution at +93 of the beta-glucuronidase gene (uidA) unique to E. coli O157:H7. An initial test panel of five E. coli and non-E. coli isolates was tested with individual primer sets (simplex assay) and all primer sets including stx1, stx2, and uidA (multiplex assay). All strains were correctly identified in both assays. Average melt temperatures (Tm's, degrees C) for PCR products were 85.42--stx1, 81.93--stx2, and 88.25--uidA in simplex assays and 85.20--stx1, 81.20--stx2, and 88.16--uidA when multiplexed. Each of the three gene targets in one multiplex reaction could be distinguished by melt curve data with significantly different Tm's. The assay was expanded to a panel of 138 isolates consisting of STEC, E. coli O157:H7, non-toxigenic E. coli, and non-E. coli isolates with melt peaks consistent with those stated above. PMID- 16271450 TI - [Validation of BREV: comparison with reference battery in 173 children with learning disorders]. AB - INTRODUCTION: The BREV battery (Battery for rapid evaluation of cognitive functions) is a tool which can be used for the rapid neuropsychological evaluation of children aged between 4 and 9 years. OBJECTIVES: After standardization (700 unaffected children) and validation by comparison with a reference battery (202 children with epilepsy), the aim of this study was further validation in 173 children with learning disorders. POPULATION AND METHODS: The study protocol included administration of the BREV, precise neuropsychological examination and evaluation of oral and written language. Statistical analysis was used to compare the findings of the BREV with those of the reference method, and the recommendations indicated by the BREV with the final diagnoses, and to define the sensitivity and the specificity of the BREV battery. RESULTS: All the correlations between BREV tests and reference tests were significant. Recommendations after the BREV were in agreement with the conclusions of the reference evaluation in 168/172 children for language, 145/173 for the psychometric evaluation. For only 4 chidren, the results of the BREV were false negative. Diagnoses corresponded in 168/173 children for oral language, in 102/110 for written language, 166/173 for praxis disorders and 157/173 for intellectual deficit. The most predictive subtests of the BREV and sensitivity and specificity of verbal and non-verbal scores were calculated. CONCLUSION: The BREV is a reliable examination, in learning disorders, to determine the most complementary investigations both in terms of language disorders and for non verbal or global learning disabilities. PMID- 16271451 TI - [Infantile choriocarcinoma: an uncommon and curable tumor]. AB - Choriocarcinoma is a malignant growth of trophoblastic cells characterized by secretion of human chorionic gonadotropin (hCG). Infantile choriocarcinoma is a very rare tumor, which is a complication of gestational choriocarcinoma and usually had very poor prognosis before chemotherapy was used. We report 1 new case as well as a review of the literature since 1945. Our case report describes the successful treatment by chemotherapy of a newborn with cerebral metastasis. Several features are important: Infantile choriocarcinoma occurs in infants aged 0 to 6 months. Anemia, hepatomegaly and hemorrhagic syndromes are the main symptoms with sometimes cerebral, cutaneous or ear-nose and throat localisations. But diagnosis can be difficult when clinical symptoms are poor. The main diagnostic criterion is a very high plasmatic or urinary level of hCG or beta-hCG in the newborn. Histological pattern is not mandatory for diagnosis. Thoracoabdominal CT scan and cerebral MRI are necessary to find metastases. Recommended treatment is chemotherapy and surgery is discussed when a tumoral residue remains. Post-treatment surveillance is based on clinical and radiological examination as well as negativation of beta-hCG. Choriocarcinoma occurring simultaneously in mother and child have been reported. Therefore it is necessary to assay maternal serum beta-hCG when infantile choriocarcinoma is disclosed and to assay serum beta-hCG in the newborn when the mother has gestational choriocarcinoma. Infantile choriocarcinoma is a very chemosensitive tumor and is thereby potentially curable. Early diagnosis is the most important prognostic factor. PMID- 16271452 TI - [Acute ethmoiditis in children, a series of 125 cases]. AB - Acute ethmoiditis are bacterial infections of ethmoid sinuses, which may spread to the orbital or the endocranial spaces. It is essential to fit the antibiotherapy to the bacteria responsible for these infections. POPULATION AND METHODS: The charts of children hospitalized from 1995 to 2003 for an acute ethmoiditis were reviewed, particularly the results of bacterial exams and the antibiotics delivered. RESULTS: Over this 9-year period, 125 children (mean age 4.5 years) were hospitalized for acute ethmoiditis. Eighty were checked for blood cultures, which were sterile in 73 cases, and in the other cases, grew Staphylococcus, S. Pneumoniae or Streptococcus pyogenes. Seric soluble antigens were absent in the 5 cases where they were looked for. Ten children had a puncture of a subperiostal abscess: it was sterile in 5 cases, Staphylococcus was found in 4 cases, S. pneumoniae in the last case. Most children received an association of cefotaxim and fosfomycine for a mean duration of 5.6 days. Thirteen per cent of the children received 3 or 4 antibiotics. DISCUSSION: It is always difficult to found the bacteria responsible for an acute ethmoiditis. In our serie as in others, the most frequent bacteria were Staphylococcus, S. pneumoniae and S. pyogenes. CONCLUSION: In view of the bacteria responsible for these infections and their antibiotic resistance, we suggest the association of cefotaxim and fosfomycin for the first line of treatment of acute ethmoiditis. PMID- 16271453 TI - [Involvement of thyroid gland at non-Hodgkin lymphoma initial diagnosis: 2 pediatric cases]. AB - Extranodal thyroid lymphomatous involvement is rare in childhood. We report here 2 children, 1 with vertical transmission-acquired human immunodeficiency virus (HIV), presenting with lymphomatous infiltration of the thyroid gland at diagnosis. One child had infra-clinical endocrine impairment and both responded well to chemotherapy. Although the cases are too scarce to be affirmative, thyroid gland involvement doesn't seem to alter the good prognosis of childhood Burkitt's lymphoma. The third child's cancer in frequency is Non-Hodgkin Lymphomas. Presenting as the initial AIDS event in 1 patient, this case report also highlights the need to systematically propose antiretroviral therapy in vertically HIV infected children. PMID- 16271454 TI - [Pediatric features of an influenza A seasonal outbreak and its burden in pediatric emergency rooms and pediatric departments]. AB - AIM OF THE STUDY: To describe epidemiological features of an outbreak of flu A in hospitalized children and to evaluate the flu's burden in pediatric and pediatric emergencies departments. POPULATION AND METHODS: Multicenter prospective study in 2 pediatric university hospitals, in Marseille, France. Nasal swabs for flu A were performed in all the febrile children admitted during winter 2003-2004. Pre and postoutbreak admissions in pediatric and pediatric emergencies departments were compared too. RESULTS: During the outbreak, 941 children were admitted and 605 were tested for flu A. Nasal swabs were positive in 111. Attack's rate was 11.8%. In these children, infants under 2 years were 66%. Respiratory forms were uncommon, although febrile seizures and digestive troubles were much frequent. The mean hospitalization duration was almost 4 days. Flu A outbreak finally increased the pediatric and pediatric emergencies departments rates of admission, of 48% and 37% respectively. CONCLUSION: Flu is frequently underestimated in children, especially in infants. Clinical forms are various. Child's vaccination is questionable in France. PMID- 16271456 TI - Neural circuits and behaviour: developmental and evolutionary perspectives. PMID- 16271455 TI - Collagen fibril morphology and organization: implications for force transmission in ligament and tendon. AB - Connective tissue mechanical behavior is primarily determined by the composition and organization of collagen. In ligaments and tendons, type I collagen is the principal structural element of the extracellular matrix, which acts to transmit force between bones or bone and muscle, respectively. Therefore, characterization of collagen fibril morphology and organization in fetal and skeletally mature animals is essential to understanding how tissues develop and obtain their mechanical attributes. In this study, tendons and ligaments from fetal rat, bovine, and feline, and mature rat were examined with scanning electron microscopy. At early fetal developmental stages, collagen fibrils show fibril overlap and interweaving, apparent fibril ends, and numerous bifurcating/fusing fibrils. Late in fetal development, collagen fibril ends are still present and fibril bundles (fibers) are clearly visible. Examination of collagen fibrils from skeletally mature tissues, reveals highly organized regions but still include fibril interweaving, and regions that are more randomly organized. Fibril bifurcations/fusions are still present in mature tissues but are less numerous than in fetal tissue. To address the continuity of fibrils in mature tissues, fibrils were examined in individual micrographs and consecutive overlaid micrographs. Extensive microscopic analysis of mature tendons and ligaments detected no fibril ends. These data strongly suggest that fibrils in mature ligament and tendon are either continuous or functionally continuous. Based upon this information and published data, we conclude that force within these tissues is directly transferred through collagen fibrils and not through an interfibrillar coupling, such as a proteoglycan bridge. PMID- 16271458 TI - Motor functions of the parietal lobe. AB - There is now general agreement that the posterior parietal cortex is part of the motor system. New data have confirmed its fundamental role in visuomotor transformations. Most interestingly, recent data showed that the inferior parietal lobule codes motor acts (such as grasping) in a specific way according to the action in which they are embedded. This particular motor organization appears to provide a neural mechanism for higher order cognitive motor functions, including understanding of intention. These functions, and peripersonal space representation, are represented in areas of the inferior parietal lobule, where visual information from both the dorsal and the ventral stream is integrated with motor information. PMID- 16271459 TI - Object memory and perception in the medial temporal lobe: an alternative approach. AB - The medial temporal lobe (MTL) includes several structures--the hippocampus, and the adjacent perirhinal, entorhinal and parahippocampal cortices--that have been associated with memory for at least the past 50 years. These components of the putative 'MTL memory system' are thought to operate together in the service of declarative memory--memory for facts and events--having little or no role in other functions such as perception. Object perception, however, is thought to be independent of the MTL, and instead is usually considered to be the domain of the ventral visual stream (VVS) or 'what' pathway. This 'textbook' view fits squarely into the prevailing paradigm of anatomical modularisation of psychological function in the brain. Recent studies, however, question this view, indicating that first, the MTL is functionally heterogeneous, and second, structures in the MTL might have a role in perception. Furthermore, the specific contributions of the individual structures within the MTL are being elucidated. These new findings indicate that it might no longer be useful to assume a strict functional dissociation between the MTL and the VVS, and that psychological functions might not be modularised in the way usually assumed. We propose an alternative approach to understanding the functions of these brain regions in terms of what computations they perform, and what representations they contain. PMID- 16271457 TI - Beetles, boxes and brain cells: neural mechanisms underlying valuation and learning. AB - Sensory cues in the environment can predict the availability of reward. Through experience, humans and animals learn these predictions and use them to guide their actions. For example, we can learn to discriminate chanterelles from ordinary champignons through experience. Assuming the development of a taste for the complex and lingering flavors of chanterelles, we therefore learn to value the same action--picking mushrooms--differentially depending upon the appearance of a mushroom. One major goal of cognitive neuroscience is to understand the neural mechanisms that underlie this sort of learning. Because the acquisition of rewards motivates much behavior, recent efforts have focused on describing the neural signals related to learning the value of stimuli and actions. Neurons in the basal ganglia, in midbrain dopamine areas, in frontal and parietal cortices and in other brain areas, all modulate their activity in relation to aspects of learning. By training monkeys on various behavioral tasks, recent studies have begun to characterize how neural signals represent distinct processes, such as the timing of events, motivation, absolute (objective) and relative (subjective) valuation, and the formation of associative links between stimuli and potential actions. In addition, a number of studies have either further characterized dopamine signals or sought to determine how such signaling might interact with target structures, such as the striatum and rhinal cortex, to underlie learning. PMID- 16271460 TI - The neural basis of smooth-pursuit eye movements. AB - Smooth-pursuit eye movements are used to stabilize the image of a moving object of interest on the fovea, thus guaranteeing its high-acuity scrutiny. Such movements are based on a phylogenetically recent cerebro-ponto-cerebellar pathway that has evolved in parallel with foveal vision. Recent work has shown that a network of several cerebrocortical areas directs attention to objects of interest moving in three dimensions and reconstructs the trajectory of the target in extrapersonal space, thereby integrating various sources of multimodal sensory and efference copy information, as well as cognitive influences such as prediction. This cortical network is the starting point of a set of parallel cerebrofugal projections that use different parts of the dorsal pontine nuclei and the neighboring rostral nucleus reticularis tegmenti pontis as intermediate stations to feed two areas of the cerebellum, the flocculus-paraflocculus and the posterior vermis, which make mainly complementary contributions to the control of smooth pursuit. PMID- 16271461 TI - Neural mechanisms of imitation. AB - Recent advances in our knowledge of the neural mechanisms of imitation suggest that there is a core circuitry of imitation comprising the superior temporal sulcus and the 'mirror neuron system', which consists of the posterior inferior frontal gyrus and adjacent ventral premotor cortex, as well as the rostral inferior parietal lobule. This core circuitry communicates with other neural systems according to the type of imitation performed. Imitative learning is supported by interaction of the core circuitry of imitation with the dorsolateral prefrontal cortex and perhaps motor preparation areas--namely, the mesial frontal, dorsal premotor and superior parietal areas. By contrast, imitation as a form of social mirroring is supported by interaction of the core circuitry of imitation with the limbic system. PMID- 16271462 TI - Time and tide in cerebellar memory formation. AB - The notion that the olivocerebellar system is crucial for motor learning is well established. In recent years, it has become evident that there can be many forms of both synaptic and non-synaptic plasticity within this system and that each might have a different role in developing and maintaining motor learning across a wide range of tasks. There are several possible molecular and cellular mechanisms that could underlie adaptation of the vestibulo-ocular reflex and eyeblink conditioning. Although causal relationships between particular cellular processes and individual components of a learned behaviour have not been demonstrated unequivocally, an overall picture is emerging that the different types and sites of cellular plasticity relate importantly to the stage of learning and/or its temporal specifics. PMID- 16271463 TI - Motor systems. PMID- 16271464 TI - Motor control and learning in altered dynamic environments. AB - Dynamic perturbations of reaching movements are an important technique for studying motor learning and adaptation. Adaptation to non-contacting, velocity dependent inertial Coriolis forces generated by arm movements during passive body rotation is very rapid, and when complete the Coriolis forces are no longer sensed. Adaptation to velocity-dependent forces delivered by a robotic manipulandum takes longer and the perturbations continue to be perceived even when adaptation is complete. These differences reflect adaptive self-calibration of motor control versus learning the behavior of an external object or 'tool'. Velocity-dependent inertial Coriolis forces also arise in everyday behavior during voluntary turn and reach movements but because of anticipatory feedforward motor compensations do not affect movement accuracy despite being larger than the velocity-dependent forces typically used in experimental studies. Progress has been made in understanding: the common features that determine adaptive responses to velocity-dependent perturbations of jaw and limb movements; the transfer of adaptation to mechanical perturbations across different contact sites on a limb; and the parcellation and separate representation of the static and dynamic components of multiforce perturbations. PMID- 16271465 TI - The basal ganglia: learning new tricks and loving it. AB - The field of basal ganglia research is exploding on every level - from discoveries at the molecular level to those based on human brain imaging. A remarkable series of new findings support the view that the basal ganglia are essential for some forms of learning-related plasticity. Other new findings are challenging some of the basic tenets of the field as it now stands. Combined with the new evidence on learning-related functions of the basal ganglia, these studies suggest that the basal ganglia are parts of a brain-wide set of adaptive neural systems promoting optimal motor and cognitive control. PMID- 16271469 TI - What does structure tell us about virus evolution? AB - Viruses are the most abundant life form and infect practically all organisms. Consequently, these obligate parasites are a major cause of human suffering and economic loss. The organization and origins of this enormous virosphere are profound open questions in biology. It has generally been considered that viruses infecting evolutionally widely separated organisms (e.g. bacteria and humans) are also distinct. However, recent research contradicts this picture. Structural analyses of virion architecture and coat protein topology have revealed unexpected similarities, not visible in sequence comparisons, suggesting a common origin for viruses that infect hosts residing in different domains of life (bacteria, archaea and eukarya). PMID- 16271466 TI - Computational motor control in humans and robots. AB - Computational models can provide useful guidance in the design of behavioral and neurophysiological experiments and in the interpretation of complex, high dimensional biological data. Because many problems faced by the primate brain in the control of movement have parallels in robotic motor control, models and algorithms from robotics research provide useful inspiration, baseline performance, and sometimes direct analogs for neuroscience. PMID- 16271470 TI - Leptostatin: a synthetic hybrid of the cytotoxic polyketides callystatin A and leptomycin B. AB - Four stereoisomeric hybrids of the polyketide natural products callystatin A and leptomycin B have been prepared by parallel synthetic routes involving chiral allenylstannane methodology. Like their natural counterparts, these hybrids exhibit nanomolar levels of cytotoxicity toward HCT-116 human colon cancer cells. PMID- 16271471 TI - Synthesis and biological evaluation of a technetium-99m(I)-tricarbonyl-labelled phenyltropane derivative. AB - A new tropane derivative was synthesized by combining a tridentate ligand, N-(2 picolylamine)-N-acetic acid (2-PAA), and a phenyltropane derivative. It was labelled with a [(99m)Tc(CO)(3)](+) moiety, resulting in the formation of two stable and neutral lipophilic isomers. Their identity was confirmed using radio LC-MS. In normal mice, no brain uptake was observed for any of the isomers and in vitro autoradiography using mouse brain sections showed no specific uptake in the striatal area. PMID- 16271472 TI - Design, synthesis and structure-activity relationships of new phosphinate inhibitors of MurD. AB - A series of new phosphinate compounds were designed and synthesized as inhibitors of the d-glutamic acid-adding enzyme (MurD) involved in peptidoglycan biosynthesis. They were tested against the MurD enzyme from Escherichia coli, allowing initial structure-activity relationships to be deduced. Two compounds had IC(50) values near 100 microM and constitute a promising starting point for further development. PMID- 16271473 TI - Stereoselective synthesis and glycosidase inhibitory activity of 3,4-dihydroxy pyrrolidin-2-one, 3,4-dihydroxy-piperidin-2-one and 1,2-dihydroxy-pyrrolizidin-3 one. AB - 3,4-Dihydroxy-pyrrolidin-2-one, 3,4-dihydroxy-piperidin-2-one and 1,2-dihydroxy pyrrolizidin-3-one have been synthesized, using a simple strategy based on the asymmetric dihydroxylation of vinylogous aminoesters and subsequent mild intramolecular cyclization. All these compounds show a partial inhibition of alpha-glucosidase, but were inactive towards other glycosidases. PMID- 16271474 TI - Synthesis and interactions of 7-deoxy-, 10-deacetoxy, and 10-deacetoxy-7 deoxypaclitaxel with NCI/ADR-RES cancer cells and bovine brain microvessel endothelial cells. AB - 7-Deoxypaclitaxel, 10-deacetoxypaclitaxel and 10-deacetoxy-7-deoxypaclitaxel were prepared and evaluated for their ability to promote assembly of tubulin into microtubules, their cytotoxicity against NCI/ADR-RES cells and for their interactions with P-glycoprotein in bovine brain microvessel endothelial cells. The three compounds were essentially equivalent to paclitaxel in cytotoxicity against NCI/ADR-RES cells. They also appeared to interact with P-glycoprotein in the endothelial cells with the two 10-deacetoxy compounds having less interaction than paclitaxel and 7-deoxypaclitaxel. PMID- 16271475 TI - Identification of the minimal protein domain required for priming activity of Munc13-1. AB - Most nerve cells communicate with each other through synaptic transmission at chemical synapses. The regulated exocytosis of neurotransmitters, hormones, and peptides occurs at specialized membrane areas through Ca2+-triggered fusion of secretory vesicles with the plasma membrane . Prior to fusion, vesicles are docked at the plasma membrane and must then be rendered fusion-competent through a process called priming. The molecular mechanism underlying this priming process is most likely the formation of the SNARE complex consisting of Syntaxin 1, SNAP 25, and Synaptobrevin 2. Members of the Munc13 protein family consisting of Munc13-1, -2, -3, and -4 were found to be absolutely required for this priming process . In the present study, we identified the minimal Munc13-1 domain that is responsible for its priming activity. Using Munc13-1 deletion constructs in an electrophysiological gain-of-function assay of chromaffin-granule secretion, we show that priming activity is mediated by the C-terminal residues 1100-1735 of Munc13-1, which contains both Munc13-homology domains and the C-terminal C2 domain. Priming by Munc13-1 appears to require its interaction with Syntaxin 1 because point mutants that do not bind Syntaxin 1 do not prime chromaffin granules. PMID- 16271476 TI - UNC-13 interaction with syntaxin is required for synaptic transmission. AB - Neurotransmitter secretion at synapses is controlled by several processes morphological docking of vesicles at release sites, priming of docked vesicles to make them fusion competent, and calcium-dependent fusion of vesicles with the plasma membrane . In worms, flies, and mice, mutants lacking UNC-13 have defects in vesicle priming . Current models propose that UNC-13 primes vesicles by stabilizing Syntaxin's "open" conformation by directly interacting with its amino terminal regulatory domain . However, the functional significance of the UNC 13/Syntaxin interaction has not been tested directly. A truncated protein containing the Munc homology domains (MHD1 and MHD2) and the carboxy-terminal C2 domain partially rescued both the behavioral and secretion defects of unc-13 mutants in C. elegans. A double mutation in MHD2 (F1000A/K1002A) disrupts the UNC 13/Syntaxin interaction. The rate of endogenous synaptic events and the amplitude of nerve-evoked excitatory post-synaptic currents (EPSCs) were both significantly reduced in UNC-13S(F1000A/K1002A). However, the pool of primed (i.e., fusion competent) vesicles was normal. These results suggest that the UNC-13/Syntaxin interaction is conserved in C. elegans and that, contrary to current models, the UNC-13/Syntaxin interaction is required for nerve-evoked vesicle fusion rather than synaptic-vesicle priming. Thus, UNC-13 may regulate multiple steps of the synaptic-vesicle cycle. PMID- 16271477 TI - On the desirability of models for inferring genome phylogenies. PMID- 16271478 TI - Low-intensity anticoagulation for stroke prevention in elderly patients with atrial fibrillation: efficacy and safety in actual clinical practice. AB - Low-intensity warfarin (INR 1.5 to 2.5) was started in 63 patients with atrial fibrillation (AF) and they were prospectively followed for 2.3 +/- 1.4 years to determine the efficacy and safety of anticoagulation for stroke prevention in actual clinical practice. Although the patients in this practice were older (76 +/- 7 years), consisted of more women (52%), and had more risk factors for stroke compared with those in clinical trials, the annual event rates of stroke and systemic embolism in this practice were comparable to those of patients receiving warfarin in clinical trials (2.0% vs. 1.4% and 0.7% vs. 0.3%). The rate of major bleeding did not significantly differ between this practice and clinical trials (0.7% vs. 1.3%). The rate of minor bleeding was significantly lower in this practice than in clinical trials (3.4% vs. 7.9%). The data suggest that low intensity anticoagulation is effective and safe for stroke prevention in elderly patients with AF at stroke risk in actual clinical practice. PMID- 16271479 TI - Ventilation according to the open lung concept attenuates pulmonary inflammatory response in cardiac surgery. AB - OBJECTIVE: Cardiac surgery with cardiopulmonary bypass (CPB) is associated with a systemic inflammatory response, which is correlated with outcome. We hypothesized that ventilation according to the open lung concept (OLC) attenuates cytokine release. METHODS: A prospective, single center randomized controlled clinical study containing 62 patients scheduled for elective coronary artery bypass graft and/or valve surgery with cardiopulmonary bypass. Before surgery, patients were randomly assigned to three groups: (1) conventional mechanical ventilation (CV), (2) OLC started after arrival on the ICU (late open lung, LOL), and (3) OLC started directly after intubation (early open lung, EOL). In both OLC groups, recruitment maneuvers were applied until PaO(2)/FiO(2)>50. The CV group received no recruitment maneuvers. Interleukin (IL)-6, IL-8, IL-10, tumor necrosis factor (TNF)-alpha, and interferon (IFN)-gamma were measured preoperatively, immediately after cessation of CPB, and 3h, 5h, 24h, 2, and 3 days after cessation of CPB. RESULTS: CPB caused a significant increase of IL-6, IL-8, and IL-10 in all groups. Thereafter, IL-8 decreased significantly more rapidly in both OLC groups compared to CV. IL-10 decreased significantly more rapidly after CPB only in the EOL group, compared with CV. Three hours after cessation of the CPB, IL-10 was already comparable with preoperative levels in the EOL group, but not in the LOL or CV group. IL-6, TNF-alpha, and IFN-gamma did not differ significantly between groups. CONCLUSIONS: OLC ventilation leads to an attenuated inflammatory response, presumably by reducing additional lung injury after cardiac surgery. Studies on cytokines after cardiac surgery should take these findings into account. PMID- 16271480 TI - Exposure of phage T7 to simulated space environment: the effect of vacuum and UV C radiation. AB - The experiment "Phage and Uracil Response" (PUR) will be accommodated in the EXPOSE facility of the International Space Station (ISS). Its objective is to examine and quantify the effect of specific space conditions on bacteriophage T7 and isolated T7 DNA thin films. In order to define the environmental and technical requirements of the EXPOSE, the samples were subjected to the Experiment Verification Test (EVT). During EVT the samples were exposed to selected space conditions: high vacuum (10(-4) to 10(-6) Pa) and UV-C radiation (254 nm) alone and in combination. Characteristic changes in the absorption spectrum, in the electrophoretic pattern of DNA/phage and the decrease of the amount of PCR products have been detected indicating the damage of isolated and intraphage T7 DNA. Intraphage DNA is more sensitive to simulated space parameters than isolated T7 DNA in thin layers as well. We obtained substantial evidence that DNA lesions accumulate throughout exposure, and the amount of damage depends on the thickness of the layers. According to our preliminary results, the damages by exposure to conditions of dehydration and UV irradiation are larger than the sum of vacuum alone, or radiation alone case, suggesting a synergistic action of space vacuum and UV radiation with DNA being the critical target. PMID- 16271481 TI - Inherited ACTH insensitivity illuminates the mechanisms of ACTH action. AB - Adrenocorticotrophin (ACTH) insensitivity is a potentially lethal inherited disorder of ACTH signalling in the adrenal. Inactivating mutations of the ACTH receptor account for approximately 25% of these cases. A second genetic cause for this syndrome has recently been identified in the MRAP gene. The MRAP protein appears to function in the trafficking and cell surface expression of the ACTH receptor, and might indicate the existence of more widespread G-protein-coupled receptor trafficking mechanisms. Molecular defects underlying other causes of ACTH insensitivity syndromes will probably contribute further to our understanding of these pathways. PMID- 16271482 TI - A consistent relationship between local white matter architecture and functional specialisation in medial frontal cortex. AB - Functionally significant landmarks in the brain do not necessarily align with local sulcal and gyral architecture in a manner that is consistent across individuals. However, the functional specialisation underlying these landmarks is strongly constrained by the connectional architecture of the region. Here, we explore this relationship in the supplementary motor area (SMA) and pre-SMA in the medial frontal cortex of the human brain. Using diffusion tensor, conventional and functional MR imaging, we find that the location of the functional boundary between SMA and preSMA is more consistent with respect to specific features of the local white matter as it approaches neocortex than with respect to the local gyral and sulcal anatomy in the region. PMID- 16271483 TI - Evaluation of EEG localization methods using realistic simulations of interictal spikes. AB - Performing an accurate localization of sources of interictal spikes from EEG scalp measurements is of particular interest during the presurgical investigation of epilepsy. The purpose of this paper is to study the ability of six distributed source localization methods to recover extended sources of activated cortex. Due to the frequent lack of a gold standard to evaluate source localization methods, our evaluation was performed in a controlled environment using realistic simulations of EEG interictal spikes, involving several anatomical locations with several spatial extents. Simulated data were corrupted by physiological EEG noise. Simulations involving pairs of sources with the same amplitude were also studied. In addition to standard validation criteria (e.g., geodesic distance or mean square error), we proposed an original criterion dedicated to assess detection accuracy, based on receiver operating characteristic (ROC) analysis. Six source localization methods were evaluated: the minimum norm, the minimum norm weighted by multivariate source prelocalization (MSP), cortical LORETA with or without additional minimum norm regularization, and two derivations of the maximum entropy on the mean (MEM) approach. Results showed that LORETA-based and MEM-based methods were able to accurately recover sources of different spatial extents, with the exception of sources in temporo-mesial and fronto-mesial regions. Several spurious sources were generated by those methods, however, whereas methods using the MSP always located very accurately the maximum of activity but not its spatial extent. These findings suggest that one should always take into account the results from different localization methods when analyzing real interictal spikes. PMID- 16271484 TI - Changes in effective connectivity during incidental and intentional perception of fearful faces. AB - The present fMRI study examined effective connectivity within an emotional network composed of three brain areas: Amygdala (AMY), Anterior Cingulate Cortex (AAC) and Orbito-Frontal (OFC) in processing fearful faces. Two tasks: an incidental perception (gender identification) and an intentional detection (effortful discrimination) task were performed by 14 and 10 young healthy volunteers, respectively. Participants were scanned while viewing fearful, neutral and ambiguous facial expressions. Effective connectivity was assessed using Structural Equation Modeling (SEM). Results show that the hypothetical network fits the experimental data for both tasks and in both hemispheres. The comparison between Tasks 1 and 2 reveals significant differences in strength and direction of the connectivity patterns for the left and to a less stringent threshold for the right hemisphere. The path coefficients analysis suggests that the fearful information generated in AMY, reaches the OFC through the ACC in incidental perception, while in intentional perception, the route followed is in a reverse direction from OFC to ACC. Our findings confirm a differential brain connectivity between incidental and intentional processing of fearful faces. PMID- 16271485 TI - A 2-year follow-up of a study to compare the efficacy of lateral wedged insoles with subtalar strapping and in-shoe lateral wedged insoles in patients with varus deformity osteoarthritis of the knee. AB - OBJECTIVE: This study was conducted in order to assess the effect of wearing a lateral wedged insole with a subtalar strap for 2 years in patients with osteoarthritis varus deformity of the knee (knee OA). DESIGN: The setting was an outpatient clinic. The efficacies of the strapped insole and a traditional shoe insert wedged insole (the inserted insole), as a positive control, were compared at the baseline and after 2 years of treatment. Randomization was performed according to birth date. The 61 female outpatients with knee OA who completed a prior 6-month study were asked to wear their respective insoles continuously as treatment during the course of the 2-year study. The femorotibial angle (FTA) was assessed by standing radiographs obtained while the subjects were barefoot and the Lequesne index of the knee OA at 2 years was compared with those at baseline in each insole group. RESULTS: There were 61 patients in the original study, but 13 patients (21.3%) did not want to wear the insole continuously and five (8.2%) withdrew for other reasons. The 42 patients who completed the 2-year study were evaluated. At the 2-year assessment, participants wearing the subtalar strapped insole (n=21) demonstrated significantly decreased FTA (P=0.015), and significantly improved Lequesne index (P=0.031) in comparison with their baseline assessments. These significant differences were not found in the group with the traditional shoe inserted wedged insole (n=21). CONCLUSION: Only those participants using the subtalar strapped insole demonstrated significant change in the FTA in comparison with the baseline assessments. If the insole with a subtalar strap maintains FTA for more than 2 years, it may restrict the progression of degenerative articular cartilage lesions of knee OA. PMID- 16271486 TI - Interaction of doxorubicin and idarubicin with red blood cells from acute myeloid leukaemia patients. AB - Doxorubicin (DOX) and idarubicin (IDA) are anthracycline antibiotics, widely used in human cancer treatment. The present study addressed the effects of these two drugs on lipid bilayer fluidity, protein conformation and microviscosity in erythrocytes from acute myeloid leukaemia patients, using electron spin resonance (ESR) spectroscopy and fluorescence measurements. Only DOX caused statistically significant changes in the parameters examined. Within 30 min of drug injection, changes were observed in the fluidity of the hydrophobic parts of the lipid bilayer and erythrocyte membrane protein conformation. These changes persisted for up to 24h. Analysis of the EPR Tempamine spectrum also showed that the microviscosity of the erythrocyte interior increased during the early stages of the drug effect. Idarubicin, in contrast, caused no identifiable change in any of the parameters studied and therefore seems to be safe for erythrocytes. We conclude that IDA is markedly less toxic than DOX to erythrocytes from acute myeloid leukaemia patients. PMID- 16271487 TI - Expression of ornithine-urea cycle enzymes in early life stages of air-breathing walking catfish Clarias batrachus and induction of ureogenesis under hyper ammonia stress. AB - The air-breathing walking catfish Clarias batrachus is a potential ureogenic teleost with having a full complement of ornithine-urea cycle (OUC) enzymes expressed in various tissues. The present study was aimed at determining the pattern of nitrogenous waste excretion in the form of ammonia-N and urea-N along with the changes of tissue ammonia and urea levels, and the expression of OUC enzymes and glutamine synthetase (GSase) in early life stages of this teleost, and further, to study the possible induction of ureogenesis in 15-day old fry under hyper-ammonia stress. The ammonia and urea excretion was visible within 12 h post-fertilization (hpf), which increased several-fold until the yolk was completely absorbed by the embryo. Although all the early developing stages were primarily ammoniotelic, they also excreted significant amount of nitrogen (N) in the form of urea-N (about 35-40% of total N). Tissue levels of ammonia and urea also increased along with subsequent developmental stages at least until the yolk absorption stage. All the OUC enzymes and GSase were expressed within 4-12 hpf showing an increasing trend of activity for all the enzymes until 350 hpf. There was a significant increase of activity of GSase, carbamyl phosphate synthetase III (CPSase III) and argininosuccinate lyase enzymes (ASL), accompanied with significant increase of enzyme protein concentration of at least two enzymes (GSase and CPSase III) in the 15-day old fry following exposure to 10 mM NH4Cl as compared to respective controls kept in water over a period of 72 h. Thus, it appears that the OUC enzymes are expressed in early life stages of walking catfish like other teleosts, but at relatively high levels and remain expressed all through the life stages with a potential of stimulation of ureogenesis throughout the life cycle as a sort of physiological adaptation to survive and breed successfully under hyper-ammonia and various other environmental-related stresses. PMID- 16271488 TI - Growth hormone- and pressure overload-induced cardiac hypertrophy evoke different responses to ischemia-reperfusion and mechanical stretch. AB - OBJECTIVE: To compare the molecular, histological, and functional characteristics of growth hormone (GH)- and pressure overload-induced cardiac hypertrophy, and their responses to ischemia-reperfusion and mechanical stretch. DESIGN: Four groups of male Wistar rats were studied: aortic banding (n=24, AB) or sham (n=24, controls) for 10 weeks, and GH treatment (n=24; 3.5mg/kg/day, GH) or placebo (n=24, controls) for 4 weeks. At 13 weeks, the rats were randomly subjected to: (i) assessment of basal left ventricular mRNA expression of sarcoplasmic reticulum calcium-ATPase (SERCA-2), phospholamban (PLB), and Na(+)-Ca(2+) exchanger (NCX) and collagen volume fraction (CVF) (Protocol A, 8 rats in each group); (ii) left ventricular no-flow ischemia with simultaneous evaluation of intracellular Ca(2+) handling and ATP, phosphocreatine (PCr) and inorganic phosphate (Pi) content (Protocol B, 12 rats in each group); or (iii) left ventricular mechanical stretch for 40 min with assessment of tumor necrosis-alpha (TNF-alpha) mRNA (Protocol C, 4 rats in each group). Protocol B and C were carried out in a Langendorff apparatus. RESULTS: In Protocol A, no difference was found as to myocardial mRNA content of Ca(2+) regulating proteins and CVF in GH animals vs controls. In contrast, in the AB group, myocardial mRNA expression of SERCA-2 and PLB was downregulated while that of NCX and CVF were increased vs. controls (p<0.05). In Protocol B, recovery of left ventricular function was significantly decreased in AB vs GH groups and controls and this was associated with 1.6-fold increase in intracellular Ca(2+) overload during reperfusion (p<0.05). Baseline ATP content was similar in the four study groups, whereas PCr and Pi was lower in AB vs GH rats and controls. However, the time courses of high energy phosphate metabolic changes did not differ during ischemia and reperfusion in the four study groups. In Protocol C, no detectable TNF-alpha mRNA level was found in the left ventricular myocardium of GH treated rats and controls at baseline, while a modest expression was noted in AB animals. Mechanical stretch resulted in de novo myocardial TNF-alpha mRNA expression in GH group and controls, which was dramatically increased in AB animals ( approximately 5-fold above baseline, p<0.001). CONCLUSIONS: The data show that cardiac hypertrophy activated by short-term GH treatment confers cardioprotection compared with pressure overload with regard to molecular and histological characteristics, and responses to ischemia-reperfusion and mechanical stretch. PMID- 16271489 TI - Expression of heme oxygenase-1 due to intracellular reactive oxygen species induced by ultrasound. AB - The present study was undertaken to elucidate the mechanism by which ultrasound induces the expression of heme oxygenase-1 (HO-1). When human lymphoma U937 cells were exposed to a 1 MHz continuous wave for 1 min, HO-1 expression examined by real-time quantitative polymerase chain reaction and immunoblotting was observed at intensities above the cavitational threshold. No induction of HO-1 expression was observed in the cells exposed for 1 min to 42 degrees C, a temperature higher than that during sonication. When a potent antioxidant, N-acetyl-l-cysteine, was added to the culture medium before or after sonication, the induction was attenuated, indicating that reactive oxygen species (ROS) are involved. However, the addition of catalase did not affect the induction, and no HO-1 was observed on the addition of pre-sonicated medium, suggesting that hydrogen peroxide due to the recombination of hydroxyl radicals generated extracellularly was not involved. The addition of free radical scavengers, glutathion-monoethyl ester, dimethyl sulfoxide and D(-)-mannitol, suppressed the induction. A decrease in mitochondrial membrane potential and the generation of superoxide were also observed in the sonicated cells, suggesting that mitochondria were the source of intracellularly generated ROS. These results indicate that superoxide secondarily generated from damaged mitochondria, not hydroxyl radicals generated in medium directly by sonication, give rise to intracellular oxidative stress inducing HO-1 expression. PMID- 16271490 TI - Application of soft tissue modelling to image-guided surgery. AB - The deformation of soft tissue compromises the accuracy of image-guided surgery based on preoperative images, and restricts its applicability to surgery on or near bony structures. One way to overcome these limitations is to combine biomechanical models with sparse intraoperative data, in order to realistically warp the preoperative image to match the surgical situation. We detail the process of biomechanical modelling in the context of image-guided surgery. We focus in particular on the finite element method, which is shown to be a promising approach, and review the constitutive relationships which have been suggested for representing tissue during surgery. Appropriate intraoperative measurements are required to constrain the deformation, and we discuss the potential of the modalities which have been applied to this task. This technology is on the verge of transition into clinical practice, where it promises to increase the guidance accuracy and facilitate less invasive interventions. We describe here how soft tissue modelling techniques have been applied to image guided surgery applications. PMID- 16271491 TI - Kiel psychotherapy project for violent offenders Towards empirically based forensic psychotherapy -- disturbance profiles and risk of recidivism among incarcerated offenders in a German prison. AB - This study reports on a psychotherapy project for violent offenders. The project was established at a German prison by the Psychiatric and Psychotherapeutic Department at the University of Kiel. The overall aim of this project is to develop a scientifically based psychotherapy programme for violent offenders. As a first step the project's systematic initial diagnostic procedures are presented. These procedures are carried out when new prisoners join the project to provide an empirical basis for the planning of therapy. Data are presented for the 60 prisoners who have so far taken part. These show high prevalences for some psychiatric disorders, namely substance related disorders (33% dependency, 55% abuse), personality disorders (58% had at least one personality disorder) and psychopathy according to Hare's criteria (13%). The data also indicate that only a basal level of self-motivation existed to take part in psychotherapy (65% precontemplative). It is apparent that violent offenders constitute a heterogeneous group as far as mental disturbances are concerned. The data suggest that a psychotherapy programme for violent offenders needs to include both targeted interventions to raise motivation and, for those with substance-related and/or personality disorders, disturbance-specific interventions. PMID- 16271492 TI - Suicide in Blantyre, Malawi (2000-2003). AB - In an attempt to identify at risk individuals, we analysed available information for individuals who committed suicide in Blantyre, Malawi. A retrospective audit of suicides autopsied at the Queen Elizabeth Central Hospital and the University of Malawi College of Medicine mortuaries between January 2000 and December 2003 was analysed by age, sex, residential location, and mode of suicide. Eighty-four suicide cases (65 males, 19 females) represented 17% of all autopsies. The major mode of suicide in Blantyre was chemical poisoning using an agricultural pesticide, accounting for 66 cases (79%)-49 males (76%), 17 females (89%). There were no cases of poisoning by therapeutic medicines, self-immolation or incised wounds. The majority of cases were from one major urban area, Limbe, and one peri urban area, Chileka. The demographics of suicide in Malawi differ from those reported for other African countries (e.g., lower proportion of females, no use of therapeutic medicine in poisoning, few gunshots). This audit highlights a need for investigations into the sale and use of agricultural pesticides. A prospective study of social and demographic factors around suicide should be undertaken to target groups at highest risk. PMID- 16271493 TI - Asymmetry of tremor intensity and frequency in Parkinson's disease and essential tremor. AB - We investigated the asymmetry of tremor intensity, frequency and frequency dispersion of Parkinsonian (PT) and essential (ET) tremor using accelerometry. Data of the more and less trembling hands were statistically elaborated. We found that tremor intensity was significantly asymmetric not only in PT but also in ET, while frequency and frequency dispersion were symmetric in ET but asymmetric in PT. We conclude that bilateral assessment of frequency related tremor parameters may be used for differentiation between ET and PT, and provides further details on the central organization of tremor generators. PMID- 16271494 TI - The effects of visual rhythms and optic flow on stride patterns of patients with Parkinson's disease. AB - This study was aimed at determining the effects of rhythmic visual cueing under changing visual conditions on stride frequency in patients with Parkinson's disease (PD; n = 21) and healthy age matched controls (n = 7) while walking at different speeds on a treadmill. Stride frequency and stride length in patients with PD as well as controls were not rigidly coupled to walking speed and could be manipulated with walking speed as well as by using spatial and temporal rhythmic visual cues. PMID- 16271495 TI - A comparison of jaw-closing and jaw-opening idiopathic oromandibular dystonia. AB - Oromandibular dystonia (OMD) is a form of focal dystonia that affects masticatory, lower facial, and lingual muscles. We compared the clinical variables and response to treatment between patients with idiopathic jaw-closure C-OMD (n = 11) and jaw-opening dystonia O-OMD (n = 12) seen in our Movement Disorders clinic over the last 10 years. The co-existence of dystonia in other regions and sensory tricks were significantly more prevalent in O-OMD (P = 0.049 and 0.03, respectively). Male gender, orobuccolingual dyskinesias (facial grimacing, lip biting, tongue dyskinesias, platysma contractions and bruxism) and better response to botulinum toxin injections were more frequent in C-OMD but remained a trend. PMID- 16271496 TI - Caregiver-burden in parkinson's disease is closely associated with psychiatric symptoms, falls, and disability. AB - The majority of care of patients with Parkinson's disease (PD) is provided by informal caregivers; their caregiving not only offers physical and emotional support for patients but also plays a large economic role and prevents early nursing home placement. In order to support caregivers in this role, it is necessary to understand the extent of caregiver-burden and factors associated with increased caregiver-burden and distress. We therefore conducted a postal survey in 123 caregivers of patients with PD to assess caregiver-burden and factors associated with it. The majority of caregivers were female (66%). Over 40% of caregivers indicated that their health had suffered as a result of caregiving, almost half had increased depression scores, and two-thirds reported that their social life had suffered. After adjustment of disease duration, there was no difference in caregiver-burden between younger and older caregivers, or between male and female caregivers. Caregiver-burden increased with increasing disability and symptoms of PD, particularly with mental health problems such as depression, hallucinations, or confusion, and with falls. Caregiver-burden scores also correlated significantly with the patients' depression and quality of life scores, and with caregivers' own satisfaction with their marital and sexual relationship. We conclude that more attention should be paid to caregivers' emotional and physical health, particularly in advancing PD with psychiatric complications and falls. These findings also demonstrate that caregiver and patient quality of life are closely linked and emphasize the importance of including caregiver-burden among the problems associated with PD in order to improve patient and caregiver lives. PMID- 16271498 TI - Bourdieu does environmental justice? Probing the linkages between population health and air pollution epidemiology. AB - The environmental justice literature faces a number of conceptual and methodological shortcomings. The purpose of this paper is to probe ways in which these shortcomings can be remedied via recent developments in related literatures: population health and air pollution epidemiology. More sophisticated treatment of social structure, particularly if based on Pierre Bourdieu's relational approach to forms of capital, can be combined with the methodological rigour and established biological pathways of air pollution epidemiology. The aim is to reformulate environmental justice research in order to make further meaningful contributions to the wider movement concerned with issues of social justice and equity in health research. PMID- 16271499 TI - Epstein-Barr virus: inhibition of apoptosis as a mechanism of cell transformation. AB - Epstein-Barr virus is a potent mitogen for human B lymphocytes and is associated with a large number of human malignancies. This large virus expresses several genes that may contribute to the transformed phenotype of infected cells and it possesses multiple strategies for the inhibition of apoptosis. The interferon inducible protein kinase PKR is an important target for the anti-apoptotic actions of the virus and its activity is regulated by the small untranslated RNA, EBER-1. This review summarizes the mechanisms of action of EBER-1 and other Epstein-Barr virus gene products in the regulation of apoptosis, and presents a model of how EBER-1 exerts its effects on PKR activity. PMID- 16271500 TI - Transforming growth factor-beta signalling in brain disorders. AB - Transforming growth factor-beta (TGF-beta) has been characterized as an injury related factor, based on the observation that it is strongly up-regulated in many acute or chronic central nervous system disorders. TGF-beta is generally thought to be neuroprotective and several mechanisms have been proposed to explain this beneficial action. For instance, TGF-beta protects neurons against the potentiating effect of tissue-type plasminogen activator on NMDA receptor mediated excitotoxicity, by up-regulating type-1 plasminogen activator inhibitor expression in astrocytes. TGF-beta has also anti-apoptotic properties, through a recruitment of a mitogen-activated protein kinase pathway and a concomitant activation of anti-apoptotic members of the Bcl-2 family. These multiple mechanisms might reflect the pleiotropic nature of TGF-beta, reinforcing the potential therapeutic value of this cytokine in several central nervous system disorders. PMID- 16271501 TI - Biofortifying crops with essential mineral elements. AB - Humans require more than 22 mineral elements, which can all be supplied by an appropriate diet. However, the diets of populations subsisting on cereals, or inhabiting regions where soil mineral imbalances occur, often lack Fe, Zn, Ca, Mg, Cu, I or Se. Traditional strategies to deliver these minerals to susceptible populations have relied on supplementation or food fortification programs. Unfortunately, these interventions have not always been successful. An alternative solution is to increase mineral concentrations in edible crops. This is termed 'biofortification'. It can be achieved by mineral fertilization or plant breeding. There is considerable genetic variation in crop species that can be harnessed for sustainable biofortification strategies. Varieties with increased mineral concentrations in their edible portions are already available, and new genotypes with higher mineral densities are being developed. PMID- 16271503 TI - Towards a neural basis of music perception. AB - Music perception involves complex brain functions underlying acoustic analysis, auditory memory, auditory scene analysis, and processing of musical syntax and semantics. Moreover, music perception potentially affects emotion, influences the autonomic nervous system, the hormonal and immune systems, and activates (pre)motor representations. During the past few years, research activities on different aspects of music processing and their neural correlates have rapidly progressed. This article provides an overview of recent developments and a framework for the perceptual side of music processing. This framework lays out a model of the cognitive modules involved in music perception, and incorporates information about the time course of activity of some of these modules, as well as research findings about where in the brain these modules might be located. PMID- 16271504 TI - Reading the hippocampal code by theta phase-locking. AB - Both the prefrontal cortex and the hippocampus are crucial for memory encoding and recall. However, it remains unclear how these brain regions communicate to exchange information. Recent findings using simultaneous recordings from the hippocampus and prefrontal cortex of the behaving rat have demonstrated that prefrontal cells' firing is phase-locked to the hippocampal theta rhythm. This suggests that phase synchronization clocked by the theta rhythm could be crucial for the communication between hippocampal and prefrontal regions. PMID- 16271505 TI - Domain-specificity and theory of mind: evaluating neuropsychological evidence. AB - Humans' unique aptitude for reasoning about mental states, known as Theory of Mind (ToM), can help explain the unique character of human communication and social interaction. ToM has been studied extensively in children, but there is no clear account of the cognitive basis of ToM in adults. Evidence from functional imaging and neuropsychology is beginning to address this surprising gap in our understanding, and this evidence is often thought to favour a domain-specific or modular architecture for ToM. We present a systematic approach to this issue for the paradigmatic case of belief reasoning, and argue that neuropsychological data provide no clear evidence for domain-specificity or modularity. Progress in understanding ToM requires new tasks that isolate potentially distinct components of this complex ability. PMID- 16271506 TI - Illusory motion reversal in tune with motion detectors. PMID- 16271507 TI - The neural correlate of (un)awareness: lessons from the vegetative state. AB - Consciousness has two main components: wakefulness and awareness. The vegetative state is characterized by wakefulness without awareness. Recent functional neuroimaging results have shown that some parts of the cortex are still functioning in 'vegetative' patients. External stimulation, such as a painful stimulus, still activates 'primary' sensory cortices in these patients but these areas are functionally disconnected from 'higher order' associative areas needed for awareness. Such studies are disentangling the neural correlates of the vegetative state from the minimally conscious state, and have major clinical consequences in addition to empirical importance for the understanding of consciousness. PMID- 16271508 TI - Feelings you can't imagine: towards a cognitive neuroscience of alexithymia. AB - Alexithymia, or 'no words for feelings', refers to an impairment of the ability to identify and communicate one's emotional state, in addition to diminished affect-related fantasy and imagery. A recent study by Mantani et al. reported reduced activation of the posterior cingulate cortex in people with alexithymia when they imagined a future happy event. This finding augments the emerging understanding of the neural basis of alexithymia, and potentially provides valuable insights into brain systems underlying normal emotion processing. PMID- 16271509 TI - Predominant expression of B7-H1 and its immunoregulatory roles in oral squamous cell carcinoma. AB - We examined cell surface expression of five B7 costimulatory molecules (B7-H1, B7 DC, B7h, CD80 and CD86) in human oral squamous cell carcinoma (SCC) lines. Most human SCC cell lines expressed various levels of B7-H1 and B7-DC. Their expression was further upregulated by interferon (IFN)-gamma stimulation. Immunohistochemical staining revealed substantial and predominant expression of B7-H1 on human primary oral SCC. A murine SCC line, NR-S1, neither expressed B7 H1 nor B7-DC, but induced B7-H1 by IFN-gamma stimulation in culture and the inoculation in vivo. Although NR-S1 tumors grew progressively in immunocompetent syngeneic mice, the administration of blocking anti-B7-Hl or anti-PD-1 mAb significantly inhibited the tumor growth, suggesting the negative regulation of host immune responses by the PD-1:B7-H1 pathway. Our results demonstrate that B7 H1 is predominantly induced on oral SCC within the B7 family molecules. A successful inhibition of tumor growth by blockade of the PD-1:B7-H1 pathway may implicate a new approach for immunotherapy of oral SCC. PMID- 16271510 TI - HBV genotype G-an odd genotype of unknown origin. PMID- 16271511 TI - CD82 metastasis suppressor gene: a potential target for new therapeutics? AB - The transmembrane glycoprotein CD82 is a member of the tetraspanin protein family and is a metastasis suppressor implicated in biological processes ranging from fusion, adhesion and migration to apoptosis and cell-morphology alterations. Downregulation of CD82 expression is associated with the advanced stages of many human cancers and correlates with the acquisition of metastatic potential. Recent studies suggest that complex mechanisms underlie CD82 loss of function, including altered transcriptional regulation, splice variant production and post translational protein modifications, and indicate a central role for CD82 in controlling metastasis as a 'molecular facilitator'. The diverse array of functions of CD82, the complexity of the regulation of CD82 and the prospects for targeting CD82 as a therapeutic approach for the treatment of a variety of metastatic cancers are discussed. PMID- 16271512 TI - A JAK2 mutation in myeloproliferative disorders: pathogenesis and therapeutic and scientific prospects. AB - Myeloproliferative disorders include several pathologies sharing the common feature of being clonal hematopoietic stem cell diseases. The molecular basis of chronic myeloid leukemia was characterized many years ago with the discovery of the t(9;22) translocation and its product the BCR-ABL oncoprotein. The recent finding of a recurrent mutation in the Janus 2 tyrosine kinase gene is a major advance in our understanding of the pathogenesis of several other myeloproliferative disorders, including polycythemia vera, essential thrombocythemia and idiopathic myelofibrosis. Although this work clearly identifies a frequent ( approximately 50%) subgroup of myeloproliferative disorders and explains most biological abnormalities described so far, it also raises the major question of how a single mutation can explain disease heterogeneity. Such a recurrent and unique mutation leading to a tyrosine kinase deregulation would make a suitable target for the development of specific therapies. PMID- 16271514 TI - FDA guidance document on monitoring delayed adverse events a good first start. PMID- 16271513 TI - IL-12 reverses anergy to T cell receptor triggering in human lung tumor associated memory T cells. AB - Memory T cells in human non-small cell lung cancer are unresponsive to progressing tumors. T cells were evaluated at the single cell level by imaging the nuclear translocation of NF-kappaB and NFAT via immunofluorescence confocal microscopy as an early measure of responsiveness to T cell receptor triggering. Little translocation of NF-kappaB or NFAT occurred in tumor-associated T cells in response to CD3+CD28 cross-linking under conditions which led to maximal translocation in normal donor peripheral blood T cells. TNF-alpha induced maximal NF-kappaB translocation in these T cells, indicating that they remain receptive to alternative signaling pathways, and pulsing with IL-12 prior to TCR triggering reversed their apparent anergy. T cells from additional chronic inflammatory microenvironments demonstrated a similar refractoriness to TCR activation, suggesting either that a common regulatory mechanism present within the microenvironment controls these cells or that with continuous antigen exposure, they remain refractory to activation via the TCR. PMID- 16271515 TI - Overcoming diabetes-induced hyperglycemia through inhibition of hepatic phosphoenolpyruvate carboxykinase (GTP) with RNAi. AB - Phosphoenolpyruvate carboxykinase (PEPCK; EC 4.1.1.32) is the rate-controlling enzyme in gluconeogenesis. In diabetic individuals, altered rates of gluconeogenesis are responsible for increased hepatic glucose output and sustained hyperglycemia. Liver-specific inhibition of PEPCK has not been assessed to date as a treatment for diabetes. We have designed a therapeutic, vector-based RNAi approach to induce posttranscriptional gene silencing of hepatic PEPCK using nonviral gene delivery. A transient reduction of PEPCK enzymatic activity (7.6 +/ 0.6 vs 9.7 +/- 1.1 mU/mg, P < 0.05) that correlated with decreased protein content of up to 50% was achieved using this strategy in diabetic mice. PEPCK partial silencing was sufficient to demonstrate lowered blood glucose (218 +/- 26 vs 364 +/- 33 mg/dl, P < 0.001) and improved glucose tolerance together with decreased circulating FFA (0.89 +/- 0.10 vs 1.44 +/- 0.11 mEq/dl, P < 0.001) and TAG (65 +/- 11 vs 102 +/- 16 mg/dl, P < 0.01), in the absence of liver steatosis or lactic acidosis. SREBP1c was down-regulated in PEPCK-silenced animals, suggesting a role for this pathway in the alterations of lipid metabolism. These data reinforce the significance of PEPCK in sustaining diabetes-induced hyperglycemia and validate liver-specific intervention at the level of PEPCK for diabetes gene therapy. PMID- 16271516 TI - Lead-induced hsp70 and hsp60 pattern transformation and leg malformation during postembryonic development in the oribatid mite, Archegozetes longisetosus Aoki. AB - The study aimed at analysing the impact of high lead concentrations on the morphological integrity and the stress protein hsp70 and hsp60 levels during postembryonic development of the oribatid mite, Archegozetes longisetosus. Independent of the treatment, the recorded hsp70 levels were far higher than the hsp60 levels in all investigated stages. There was a tendency towards lower hsp70 and hsp60 levels with proceeding development (deutonymph>tritonymph>adult) in untreated animals. Both the hsp70 and hsp60 levels in all investigated quiescent stages prior to moult were higher than in the corresponding active stages independent from lead exposure. Continuous lead treatment from the larval stage onwards caused malformation of the 4th pair of legs and, in parallel, a shift to elevated hsp70 (but not hsp60) levels in all subsequent stages, compared to controls. Neither effects occurred when continuous lead treatment started later in development. In this case, elevated hsp60 levels could particularly be found in those stages respectively following the initially exposed stage. The hsp70 response became obvious even quicker in tritonymphs and adults, where hsp70 level peaks could be observed right in those stages the lead exposure had started in. PMID- 16271517 TI - JANUS under stress--role of JAK/STAT signaling pathway in vascular diseases. AB - They were more than just another kinases (JAK), when they were first described in the late 80s and named JAK kinases. The mandatory role of this novel family of dual active janus kinases (JAK) and their substrates the signal transducers and activators of transcription (STAT) was demonstrated in mice who died during embryogenesis when lacking a functional allele, e.g. that of JAK2. Initially, the JAK/STAT signaling pathway was discovered as the primary mediator of intracellular signaling induced by interferon in hematopoietic and immune cells. Nowadays, it is well accepted that JAK kinases and STAT proteins are constitutively expressed in the vessel wall in a cell type specific manner and transfer intracellular signaling events of various receptor families, e.g. that of cytokines, growth factors and vasoactive peptides such as angiotensin II (Ang II) or endothelin. The potential impact of the JAK/STAT signaling pathway on cardiovascular pathophysiology and disease development arise from reports describing that JAKs may bind directly to the angiotensin II type I (AT(1)) receptor, thereby enhancing their phosphorylation in various cell types of the vessel wall. More interestingly, these signaling events are modulated by NAD(P)H oxidase-derived superoxide anions which directly phosphorylate JAK2 and thereby control JAK2 activity. A potential impact was also described for atherosclerotic plaque development in which the activation of JAKs and STATs seems to be critical. Based on these observations, we here review the role of the JAK/STAT signaling pathways as critical regulator for cardiovascular disease development, i.e. atherosclerotic plaque progression or the manifestation of arterial hypertension. PMID- 16271518 TI - The BMP type II receptor is located in lipid rafts, including caveolae, of pulmonary endothelium in vivo and in vitro. AB - Polymorphic mutations in the Bone Morphogenetic Protein type II receptor (BMPrII) gene have been implicated in the development of familial primary pulmonary hypertension (PPH) however, the role BMPrII mutations play in the development of PH has not yet been elucidated. Endothelial caveolae are an important domain of hemodynamics containing eNOS, the serotonin transporter, and endothelin receptors. In this study we show by standard immunohistochemistry (IHC) that BMPrII is widely distributed in the vasculature of the rat lung, and more specifically distributed to both apical and basal membranes of the arteriolar endothelium by fluorescent IHC. We also examined compartmentalization of BMPrII in lipid fractions of plasma membranes isolated by silica based extraction from human pulmonary artery endothelial cells and rat lung endothelium. Density gradient centrifugation demonstrated BMPrII in separate caveolin-1 (cav-1) and non-cav-1 lipid rich fractions. Electron microscopy co-localized cav-1 and BMPrII in flask shaped membrane fragments. Three-dimensional fluorescence microscopy demonstrated BMPrII in discrete membrane foci, a portion of which were co localized with cav-1, as well as in Golgi. Our findings indicate that BMPrII is located within lipid-dense fractions of pulmonary endothelial cell membranes with a portion present in caveolae suggesting potential dynamic regulatory structural relationships. PMID- 16271519 TI - Editorial letter. PMID- 16271521 TI - Controlled clinical trials in cystic fibrosis--are we doing better? AB - We have previously reported the time trends, design and interventions in randomised controlled trials (RCTs) in cystic fibrosis (CF) from 1961 through 1997 [Cheng K, Smyth RL, Motley J, O'Hea U, Ashby D, Randomised controlled trials in cystic fibrosis (1966-1997) categorized by time, design, and intervention. Pediatr Pulmonol 2000, 29:1-7.]. We maintain an ongoing register of all RCTs and controlled clinical trials (CCTs) in CF and have noted that in the five years since 1997 there has been a 48% increase in published trials. We aimed to assess whether this increase has been associated with an improvement in design quality. All RCTs and CCTs from 1961-2002 were assessed. Two epochs were then compared, 1961-1997 and 1998-2002. For each trial we recorded the design, participant numbers and the intervention studied. 261 trials in 1998-2002 were compared with 544 trials in 1961-1997. Comparing the two epochs a similar proportion of trials were parallel, double-blind and placebo controlled; also the median number of participants was similar. In the later epoch 25% of trials were multicentre, compared with 11% previously. Whilst this recent increase in clinical trials in CF is welcome, this has not been associated with improvements in quality. The trend for an increasing proportion of trials to be multicentre is encouraging. There are however, still deficiencies in the design of clinical trials in CF. PMID- 16271520 TI - Longevity-associated mitochondrial DNA 5178 C/A polymorphism is associated with fasting plasma glucose levels and glucose tolerance in Japanese men. AB - Mitochondrial DNA 5178 cytosine/adenine (Mt5178 C/A) polymorphism is reportedly associated with longevity in the Japanese population, and the Mt5178A genotype may resist the onset of type 2 diabetes. To investigate whether Mt5178 C/A polymorphism is associated with glucose tolerance, we conducted a cross-sectional study using the 75-g oral glucose tolerance test (OGTT) in which non-diabetic Japanese male subjects were classified into three subgroups by body mass index (BMI): BMI<22 (n=91); 22< or =BMI<25 (n=138); and BMI> or =25 (n=67). The frequency of Mt5178A was significantly lower among 'BMI<22' subjects exhibiting impaired fasting glucose and impaired glucose tolerance than among those with normal glucose tolerance. In the 'BMI<22' group, fasting plasma glucose (FPG) levels and plasma glucose levels at 60 and 120 min after glucose load (OGTT-1h and OGTT-2h, respectively) were significantly lower in the Mt5178A genotype than in the Mt5178C genotype. After adjusting for age, BMI, habitual smoking, habitual drinking and family history of diabetes, FPG levels and OGTT-2h levels were still significantly lower in the Mt5178A genotype than in the Mt5178C genotype. However, after adjusting for covariates, in both the '22< or =BMI<25' and 'BMI> or =25' groups, FPG levels were significantly higher in the Mt5178A genotype than in the Mt5178C genotype. Differences in the effect of alcohol consumption on FPG levels and glucose tolerance between the Mt5178 C/A genotypes were observed. The present results suggest that Mt5178 C/A polymorphism may be associated with FPG levels and glucose tolerance in middle-aged Japanese men. PMID- 16271522 TI - Protein kinases as targets for antimalarial intervention: Kinomics, structure based design, transmission-blockade, and targeting host cell enzymes. AB - The surge of interest in protein kinases as targets for chemotherapeutic intervention in a number of diseases such as cancer and neurodegenerative disorders has stimulated research aimed at determining whether enzymes of this class might also be considered as targets in the context of diseases caused by parasitic protists. Here, we present an overview of recent developments in this field, concentrating (i) on the benefits gained from the availability of genomic databases for a number of parasitic protozoa, (ii) on the emerging field of structure-aided design of inhibitors targeting protein kinases of parasitic protists, (iii) on the concept known as transmission-blockade, whereby kinases implicated in the development of the parasite in their arthropod vector might be targeted to interfere with disease transmission, and (iv) on the possibility of controlling parasitic diseases through the inhibition of host cell protein kinases that are required for the establishment of infection by the parasites. PMID- 16271523 TI - Extracellular post-translational modifications of collagen are major determinants of biomechanical properties of fetal bovine cortical bone. AB - Mechanical behavior of bone depends on its mass and architecture, and on the material properties of the matrix, which is composed of a mineral phase and an organic component mainly constituted of type I collagen. Mineral accounts largely for the stiffness of bone, whereas type I collagen provides bone its ductility and toughness, i.e., its ability to undergo deformation and absorb energy after it begins to yield. The molecular mechanisms underlying the effect of alterations in type I collagen on bone mechanical properties are unclear. We used an in vitro model of fetal bovine cortical bone specimens (n = 44), where the extent of type I collagen cross-linking was modified by incubation at 37 degrees C for 0, 60, 90 and 120 days, keeping constant the architecture and the mineral content. At each incubation time, the following parameters were determined: (1) the bone concentration of enzymatic (pyridinoline; PYD and deoxypyridinoline, DPD) and non enzymatic (pentosidine) crosslinks by HPLC, (2) the extent of aspartic acid isomerization of the type I collagen C-telopeptide (CTX) by ELISA of native (alpha CTX) and isomerized (beta CTX) forms, (3) the mineral density by DXA, (4) the porosity by micro-computed tomography and (5) the bending and compressive mechanical properties. Incubation of bone specimens at 37 degrees C for 60 days increased the level (per molecule of collagen) of PYD (+98%, P = 0.005), DPD (+42%, P = 0.013), pentosidine (+55-fold, P = 0.005), and the degree of type I collagen C-telopeptide isomerization (+4.9-fold, P = 0.005). These biochemical changes of collagen were associated with a 30% decrease in bending and compressive yield stress and a 2.5-fold increase in compressive post-yield energy absorption (P < 0.02 for all), with no significant change of bone stiffness. In multivariate analyses, the level of collagen cross-linking was associated with yield stress and post-yield energy absorption independently of bone mineral density, explaining up to 25% of their variance. We conclude that the extent and nature of collagen cross-linking contribute to the mechanical properties of fetal bovine cortical bone independently of bone mineral density. PMID- 16271524 TI - SUMOylation and PPARgamma: wrestling with inflammatory signaling. AB - The molecular mechanisms whereby PPARgamma inhibits inflammatory gene expression in macrophages are poorly understood. In a recent Nature paper, provide a new model for trans-repression in which ligand-dependent SUMOylation of PPARgamma results in its recruitment to the promoters of inflammatory genes where it inhibits transcription by preventing clearance of corepressor complexes. PMID- 16271525 TI - Pulling in more fat. AB - In obesity, skeletal muscle accumulates triglyceride. Recent work from Hulver and colleagues (2005) in the October issue of Cell Metabolism implicates stearoyl-CoA desaturase as part of the underlying molecular mechanism. PMID- 16271526 TI - Flies on steroids: the interplay between ecdysone and insulin signaling. AB - In the fruit fly Drosophila melanogaster, the insulin and ecdysone signaling pathways have long been known to regulate growth and developmental timing, respectively. Recent findings reveal that crosstalk between these pathways allows coordination of growth and developmental timing and thus determines final body size. PMID- 16271527 TI - Enlightening the adrenal gland. AB - The secretion of glucocorticoid hormones is tightly regulated by the circadian clock and by negative humoral feedback loops, both acting on the hypothalamic pituitary gland-adrenal axis. However, a new study Ishida et al., 2005 [this issue of Cell Metabolism) shows that light can influence the adrenal's glucocorticoid output by a more direct pathway. PMID- 16271528 TI - SIR2 calls upon the ER. AB - Resveratrol induces longevity in C. elegans through the action of SIR2. Recently published work shows that resveratrol induces genes of the unfolded protein stress response of the endoplasmic reticulum. Paradoxically, these stress genes are repressed by SIR2, suggesting that resveratrol increases life span by inhibiting this SIR2 action. PMID- 16271529 TI - Rb and p107 regulate preadipocyte differentiation into white versus brown fat through repression of PGC-1alpha. AB - The Rb family, Rb, p107, and p130, play important roles in cell cycle control and cellular differentiation, and Rb has been suggested to regulate adipocyte differentiation. We report here that mice lacking p107 displayed a uniform replacement of white adipose tissue (WAT) with brown adipose tissue (BAT). Mutant WAT depots contained mutilocular adipocytes that expressed elevated levels of PGC 1alpha and UCP-1 typical of BAT. WAT from p107-/- mice contained markedly elevated numbers of adipogenic precursors that displayed downregulated expression of pRb. Consistent with the hypothesis that pRb is required for adult adipocyte differentiation, Cre-mediated deletion of Rb in adult primary preadipocytes blocked their differentiation into white adipocytes. Importantly, pRb was observed to bind the PGC-1alpha promoter and repress transcription. Therefore, p107 and pRb regulate PGC-1alpha expression to control the switch between white and brown adipocyte differentiation from a common pool of presumptive adult progenitors in fat tissue. PMID- 16271530 TI - Light activates the adrenal gland: timing of gene expression and glucocorticoid release. AB - Light is a powerful synchronizer of the circadian rhythms, and bright light therapy is known to improve metabolic and hormonal status of circadian rhythm sleep disorders, although its mechanism is poorly understood. In the present study, we revealed that light induces gene expression in the adrenal gland via the suprachiasmatic nucleus (SCN)-sympathetic nervous system. Moreover, this gene expression accompanies the surge of plasma and brain corticosterone levels without accompanying activation of the hypothalamo-adenohypophysial axis. The abolishment after SCN lesioning, and the day-night difference of light-induced adrenal gene expression and corticosterone release, clearly indicate that this phenomenon is closely linked to the circadian clock. The magnitude of corticostereone response is dose dependently correlated with the light intensity. The light-induced clock-dependent secretion of glucocorticoids adjusts cellular metabolisms to the new light-on environment. PMID- 16271531 TI - Unexpected role of ceruloplasmin in intestinal iron absorption. AB - Ferroxidases are essential for normal iron homeostasis in most organisms. The paralogous vertebrate ferroxidases ceruloplasmin (Cp) and hephaestin (Heph) are considered to have nonidentical functions in iron transport: plasma Cp drives iron transport from tissue stores while intestinal Heph facilitates iron absorption from the intestinal lumen. To clarify the function of Cp, we acutely bled Cp-/- mice to stress iron homeostasis pathways. Red cell hemoglobin recovery was defective in stressed Cp-/- mice, consistent with low iron availability. Contrary to expectations, iron was freely released from spleen and liver stores in Cp-/- mice, but intestinal iron absorption was markedly impaired. Phlebotomy of wild-type mice caused a striking shift of Cp from the duodenal epithelium to the underlying lamina propria, suggesting a critical function of Cp in basolateral iron transport. Regulated relocalization of intestinal Cp may represent a fail-safe mechanism in which Cp shares with Heph responsibility for iron absorption under stress. PMID- 16271532 TI - Portal sensing of intestinal gluconeogenesis is a mechanistic link in the diminution of food intake induced by diet protein. AB - Protein feeding is known to decrease hunger and subsequent food intake in animals and humans. It has also been suggested that glucose appearance into portal vein, as occurring during meal assimilation, may induce comparable effects. Here, we connect these previous observations by reporting that intestinal gluconeogenesis (i.e., de novo synthesis of glucose) is induced during the postabsorptive time (following food digestion) in rats specifically fed on protein-enriched diet. This results in glucose release into portal blood, counterbalancing the lowering of glycemia resulting from intestinal glucose utilization. Comparable infusions into the portal vein of control postabsorptive rats (fed on starch-enriched diet) decrease food consumption and activate the hypothalamic nuclei regulating food intake. Similar hypothalamic activation occurs on protein feeding. All these effects are absent after denervation of the portal vein. Thus, portal sensing of intestinal gluconeogenesis may be a novel mechanism connecting the macronutrient composition of diet to food intake. PMID- 16271534 TI - Introduction. PMID- 16271533 TI - Dual role of the coactivator TORC2 in modulating hepatic glucose output and insulin signaling. AB - Under fasting conditions, the cAMP-responsive CREB coactivator TORC2 promotes glucose homeostasis by stimulating the gluconeogenic program in liver. Following its nuclear translocation in response to elevations in circulating glucagon, TORC2 regulates hepatic gene expression via an association with CREB on relevant promoters. Here, we show that, in parallel with their effects on glucose output, CREB and TORC2 also enhance insulin signaling in liver by stimulating expression of the insulin receptor substrate 2 (IRS2) gene. The induction of hepatic IRS2 during fasting appears critical for glucose homeostasis; knockdown of hepatic IRS2 expression leads to glucose intolerance, whereas hepatic IRS2 overexpression attenuates the gluconeogenic program and reduces fasting glucose levels. By stimulating the expression of IRS2 in conjunction with gluconeogenic genes, the CREB:TORC2 pathway thus triggers a feedback response that limits glucose output from the liver during fasting. PMID- 16271535 TI - Experience with hepatitis A and B vaccines. AB - The lengthy history of efforts to understand the pathogenesis and means of preventing and controlling both hepatitis A and B is noteworthy for many exceptional scientific achievements. Among these are the development of vaccines to prevent the spread of infection through induction of active immunity to hepatitis A virus (HAV) and hepatitis B virus (HBV). The first plasma-derived hepatitis B vaccine was licensed in the United States in 1981 and was replaced by recombinant hepatitis B vaccines in 1986 and 1989. Vaccines to prevent HAV infection were licensed in the United States in 1995 and 1996. Subsequently, combination vaccines that included both hepatitis A and B vaccine components, or the hepatitis B component in combination with other commonly administered vaccines, were licensed in the United States. Despite significant reductions in hepatitis-related morbidity and mortality that have resulted from widespread use of these vaccines, vaccine-preventable morbidity and mortality still occur. The purposes of this article are to review clinical trial and other experience with hepatitis A and B vaccines in healthy individuals as well as in those with chronic liver disease, infected with the human immunodeficiency virus, or requiring hemodialysis; describe the impact that these vaccines and national recommendations for vaccination have had on reducing the incidence of HAV and HBV infection; and recommend expansion of these recommendations to include universal vaccination of adults as a means of further reducing the burden of viral hepatitis. PMID- 16271536 TI - Evaluating existing recommendations for hepatitis A and B vaccination. AB - Hepatitis A and hepatitis B vaccines are now widely available throughout the United States, Europe, and most of the rest of the world. A recombinant hepatitis B vaccine was licensed in the United States in 1986, followed by hepatitis A vaccine in 1995. Official recommendations from the Advisory Committee on Immunization Practices were published for hepatitis B vaccine in 1990, with revisions in 1991, and for hepatitis A vaccine in 1999. Unfortunately, the recommendations are not closely followed, and an insufficient number of persons who are at risk receive these vaccines. In addition, the available recommendations may not cover all who are at risk for infection. This article reviews existing recommendations for the use of hepatitis A and hepatitis B vaccines and offers suggestions for situations in which additional recommendations may be useful. The rationale for revising and expanding the existing recommendations is based on questions raised about the value of risk based recommendations, data demonstrating increasing rates of hepatitis A and B infections in specific age groups, and the failure to protect 50% of individuals who become infected with vaccine-preventable hepatitis. PMID- 16271537 TI - Acute hepatitis A and B in patients with chronic liver disease: prevention through vaccination. AB - Retrospective and prospective studies have demonstrated that the occurrence of acute hepatitis A in patients with chronic liver disease is associated with higher rates of morbidity and mortality than in previously healthy individuals with acute hepatitis A. The mortality associated with acute hepatitis A may be particularly high in patients with preexisting chronic hepatitis C. Although acute hepatitis B in patients with preexisting chronic liver disease is less well studied, worse outcomes than in previously healthy individuals are apparent. However, numerous studies convincingly demonstrate that chronic hepatitis B virus coinfection with hepatitis C virus (or hepatitis D virus) is associated with an accelerated natural history of liver disease and worse outcomes. These observations led to studies that demonstrated the safety and efficacy of hepatitis A and hepatitis B vaccination in patients with mild-to-moderate chronic liver disease. Hepatitis A and B vaccination is less effective in patients with advanced liver disease, especially after decompensation, such as in patients awaiting liver transplantation, and in liver transplant recipients. The emerging lower rates of inherent immunity in younger individuals, higher morbidity and mortality of acute hepatitis A or B superimposed on chronic liver disease, and greater vaccine efficacy in milder forms of chronic liver disease suggest that it is a reasonable policy to recommend hepatitis A and B vaccination in patients early in the natural history of chronic liver disease. PMID- 16271538 TI - Screening for hepatitis A and B antibodies in patients with chronic liver disease. AB - Chronic liver disease (CLD) is highly prevalent, and hepatitis C is one of the leading causes. Acute hepatitis A or B in patients with chronic hepatitis C can lead to more severe hepatic injury and a higher fatality rate than in patients without hepatitis C. Thus, the Advisory Committee on Immunization Practices (ACIP) of the Centers for Disease Control and Prevention and the World Health Organization recommend that persons with CLD be vaccinated against hepatitis A virus (HAV), and the ACIP and the National Institutes of Health recommend vaccination against both HAV and hepatitis B virus (HBV) in patients with chronic hepatitis C. Because coinfection with HAV or HBV in patients with chronic hepatitis C or CLD is common, antibody screening prior to hepatitis A or B vaccination can identify patients who are already immune to these viruses and thus do not need to be vaccinated. Selective hepatitis A vaccination (i.e., vaccination of patients who test negative for either HAV antibody immunoglobulin G or total antibodies to HAV) is most cost-effective in areas where the local prevalence of hepatitis A is higher than the national prevalence and in populations with higher background rates of HAV exposure compared with the general population, such as older adults, foreign-born patients, African Americans, and persons with CLD or hepatitis C. Although not usually recommended for healthy adults or those with compensated CLD because of virtually 100% postvaccination seroconversion, serologic testing after hepatitis A vaccination is recommended in patients with decompensated or advanced end-stage liver disease because of the much lower seroconversion rates in these patients. Selective vaccination against HBV in patients with CLD or hepatitis C is also recommended. Testing for hepatitis B surface antigen (HBsAg) and antibodies to HBsAg (anti HBs) is considered the most efficient and reasonably cost-effective method to screen for hepatitis B serologic markers because HBsAg identifies individuals with both acute and chronic HBV infection, and anti-HBs identify those who are immune secondary to vaccination or past infection. Testing for antibodies to hepatitis B core antigen is needed to further distinguish between immunity due to vaccination and immunity due to past infection, but it is not recommended as the only screening test for HBV immunity. Postvaccination testing for hepatitis B seroconversion is recommended in all patients with CLD, especially in those with more advanced disease, because the rate of seroconversion is generally lower than in healthy adults. If patients with CLD are not adequately protected after a standard course of hepatitis B vaccination, a repeat course of vaccination using the standard schedule or an accelerated schedule (days 0, 7, and 21) should be considered. PMID- 16271539 TI - Prevention of hepatitis B in nonresponders to initial hepatitis B virus vaccination. AB - Although vaccination against hepatitis B virus (HBV) is highly successful, 5% to 10% of individuals do not experience a response with an adequate antibody level to hepatitis B surface antigen (anti-HBs). Contributing causes for nonresponse to the vaccine are genetic predisposition, immunosuppression, and certain chronic illnesses. The distinction between true nonresponse (after adequate immunization) and waning anti-HBs levels is important. The latter is not uncommon in populations in areas of the world with low endemicity for HBV infection. Data from subjects with waning anti-HBs levels show that immunologic memory may still protect these individuals against acute HBV infection or may prevent chronic infection with HBV for < or =10 years after immunization. Recent reports from Asia and Alaska describe cases of chronic HBV infection 15 years after immunization in subjects who have very low levels of anti-HBs. Thus, nonresponders or those with waning immunity who may be at risk of HBV infection in subsequent years may require a booster dose. Clinical algorithms to reimmunize nonresponders have been described and are discussed in this article. Experimental hepatitis B vaccines have shown some promise in nonresponders but are not commercially available in the United States. PMID- 16271540 TI - Overcoming obstacles to immunization in patients with chronic liver disease. AB - The importance of vaccination to protect against hepatitis A virus (HAV) and hepatitis B virus (HBV) infections in patients with chronic liver disease has been established. However, in this population, a number of obstacles can interfere with appropriate and timely hepatitis immunization. The costs of hepatitis A and B vaccine series are out of reach for many uninsured patients. Many private and government-sponsored insurance programs do not routinely cover these vaccinations for patients with chronic liver disease. Varying recommendations by government and national organizations, such as the Centers for Disease Control and Prevention (CDC) and the American Association for the Study of Liver Diseases (AASLD), may lead to uncertainty and inconsistent vaccination practices. Because of the need for multiple office visits for prescreening assessment and vaccine administration, patient adherence can be an issue as well. Improved coverage of vaccines by government and third-party health plans is needed, as are uniform guidelines regarding the vaccination of patients with chronic liver disease. Providers should counsel such patients about the serious health risks incurred by infection with HAV or HBV and encourage vaccination in these patients. A combination of interventions can be used to facilitate timely and appropriate vaccination against hepatitis and to improve the affordability of vaccination for patients with chronic liver disease. PMID- 16271541 TI - Changing travel-related global epidemiology of hepatitis A. AB - Hepatitis A is highly endemic in many emerging cultures. Despite the availability of safe and effective vaccines and some improvements in sanitation in developing countries, hepatitis A remains a significant cause of morbidity for nonimmune travelers visiting such destinations. All are at risk, including short-term vacationers or business travelers who stay in deluxe accommodations. This may have considerable implications on public health. Hepatitis A vaccination programs for travelers have not proven to be effective, since many visitors to destinations at risk (e.g., Mexico) fail to consult health professionals prior to departure. Because 50% of the US population has an anticipated lifetime risk for exposure, universal immunization against hepatitis A should be considered. PMID- 16271542 TI - United States epidemiology of hepatitis A: influenced by immigrants visiting friends and relatives in Mexico? AB - Among the industrialized nations, the United States annually receives the greatest number of immigrants as permanent residents. Immigrants from Mexico have represented the largest segment of the foreign-born population in recent decades, and continued growth of Mexican immigration is predicted for the decades ahead. The changing demographics of this population, including the emergence of new immigrant growth centers, will influence the future epidemiology of hepatitis A virus transmission in the United States. Travel home to the place of origin to visit friends and relatives (VFR) by both newly arrived and established Mexican immigrants constitutes a new group of travelers that now include intergenerational family units. Asymptomatic pediatric travelers-who acquire hepatitis A abroad and are infectious on return to American communities contribute to the silent transmission of hepatitis A to playmates, caretakers, and contacts in households, daycare facilities, and elementary schools. Considering the expanded geographic distribution of Mexican immigrant settlement, the predicted increased diversity of pediatric populations in the United States over time, and the continued growth of VFR travel, a universal pediatric vaccine recommendation for hepatitis A immunization can help to prevent hepatitis A transmission in this country in the future. PMID- 16271543 TI - Hepatitis A vaccine in the last-minute traveler. AB - Current recommendations state that travelers should receive hepatitis A vaccine 2 to 4 weeks before departure. Such recommendations, however, may dissuade last minute travelers from receiving the vaccine. A preponderance of evidence exists to support hepatitis A vaccination of the imminent-departure traveler and therefore suggests that these guidelines merit reconsideration. In examining this issue, one of the most important elements to determine is the amount of time required for seroconversion following vaccination. Clinical trials of hepatitis A vaccines measured antibody response at 2 and 4 weeks after vaccination. However, studies investigating early seroconversion found that the vast majority of vaccinees develop antibodies within 2 weeks of vaccination, some as early as 12 days after vaccination. This is relevant information, given that the hepatitis A virus has an average incubation period of 28 days. Seroconversion is predicated on achieving a "protective" antibody level. However, levels of antibody considered protective remain debatable. Evidence suggests that clinical disease does not occur at antibody levels lower than those currently accepted as protective. Furthermore, hepatitis A vaccine has been proved effective in controlling outbreaks worldwide. Research data show that a single dose of vaccine can halt outbreaks if an adequate number of susceptible individuals are vaccinated. Information from rapid-outbreak control studies and those assessing postexposure administration of hepatitis A vaccine suggest that late vaccination provides a significant degree of protection. For these reasons, hepatitis A vaccine may be administered at any time before departure because it will still provide travelers with protection. PMID- 16271544 TI - Travel-related hepatitis B: risk factors and prevention using an accelerated vaccination schedule. AB - Rates of global travel and tourism are increasing dramatically, especially to regions with medium or high endemicity for hepatitis A and B, such as Asia, Africa, Latin America, and the Middle East. International travelers to these areas should be protected against both hepatitis A and B, regardless of their anticipated length of stay. However, many travelers depart within weeks of planning their trip (too late to complete the accelerated 0-, 1-, 2-month regimen for hepatitis B), and a majority of those traveling depart without being vaccinated. Although extended-stay travelers are at high risk for hepatitis B, short-stay travelers also are at risk. The most commonly encountered risk factors for travel-related hepatitis B are casual sexual activity with a new partner, medical and dental care abroad, and in the expatriate community, adoption of children who are hepatitis B carriers. Although efficacy studies of accelerated schedules for hepatitis B immunization have not been conducted, the results of immunogenicity studies in healthy volunteers who received an accelerated, 3-dose regimen on a 0-,7-, and 21-day schedule suggest that excellent, rapid, and long term protection will be conferred. More data are needed to assess the efficacy of accelerated schedules in persons aged >40 years and to determine whether a fourth dose of hepatitis B vaccine is needed in all age groups. PMID- 16271545 TI - Hepatitis A and B immunization in persons being evaluated for sexually transmitted diseases. AB - Sexual transmission accounts for the majority of hepatitis B virus (HBV) infections in industrialized countries. Hepatitis A virus (HAV) can be transmitted by sexual practices that involve fecal-oral exposure. Both infections are disproportionately frequent in men who have sex with men (MSM). Routine immunization against HBV is recommended for MSM and for persons being evaluated or treated for sexually transmitted diseases (STDs), and HAV immunization is advised for MSM and for other persons at risk who are commonly seen in STD care settings, such as users of illegal drugs. However, numerous attitudinal and structural barriers interfere with routine immunization in persons at risk for sexual acquisition of HAV and HBV. Substantial success has been documented in vaccinating persons at risk in public STD clinics and other settings; however, at a national level, efforts to achieve desired immunization rates have largely failed. Until universal childhood immunization produces a largely immune adult population, the universal vaccination of adults-as a supplement to the current risk-based approaches-may be worthwhile to achieve immunization of persons at risk for sexual transmission of HBV. PMID- 16271546 TI - Hepatitis A and B immunizations of individuals infected with human immunodeficiency virus. AB - All persons at risk for infection with human immunodeficiency virus (HIV) types 1 and 2, including men who have sex with men, those with multiple heterosexual contacts, abusers of illegal injection drugs, and persons frequently exposed to blood and blood products, are also at high risk for hepatitis A virus (HAV) and acute and chronic hepatitis B virus (HBV) infections. HIV can prolong the duration and increase the level of HAV viremia and augment HAV-related liver abnormalities. HIV also magnifies HBV viremia and the risk of HBV reactivation, chronic active HBV infection, cirrhosis, and death. Because of these concerns, hepatitis A vaccination is recommended for all HIV-positive/HAV seronegative persons, with 2 standard doses given 6 to 12 months apart. Immune response to hepatitis A vaccines is excellent, even in moderately immune-suppressed individuals. Hepatitis B vaccination is also recommended for all HIV-positive persons lacking prior immunity. However, immune reactivity to hepatitis B vaccines is frequently suboptimal in terms of patients' rate of response, antibody titer, and durability. Relatively high CD4+ T-cell counts (> or =500/mm3) and low levels of HIV viremia (<1,000 RNA genome copies/mL plasma) are necessary to ensure adequate hepatitis B vaccine response. Higher hepatitis B vaccine doses, prolongation of the vaccination schedule, or both, as prescribed for many patients with non-HIV-related immune deficiencies, may be considered initially. Revaccination should be instituted if postvaccination titers of antibodies to hepatitis B surface antigen are <10 mIU/mL (<10 IU/L). Nonresponders may also react to a subsequent vaccine course if CD4+ T-cell counts rise to 500/mm3 following institution of highly active antiretroviral therapy; vaccine adjuvant trials are under way. Universal, age-based immunization of all young and middle-aged adults appears to be the most comprehensive way of protecting all populations who are at high risk. PMID- 16271547 TI - A call to immunize the correctional population for hepatitis A and B. AB - An increasing number of the 12 million people who pass through jails and prisons in the United States each year are infected with hepatitis B or C. Many have routinely engaged in sexual and drug-use behaviors that put them and their contacts at risk for infection, and many are likely to return to these behaviors on their release, further spreading the infection. Thus, the prevalence of hepatitis infections among the incarcerated population is an enormous health problem not only for the correctional system but also for the community at large. There is no federal mandate for screening members of the incarcerated population who may be at risk or those who are already infected. Public health and correctional authorities must join forces to offer educational programs, prevention services, and intervention strategies to offenders, many of whom have their first contact with the healthcare system when they enter a correctional facility. Safe and effective vaccines for hepatitis A and B have been available for >11 years and, despite the challenges of conducting an immunization program for this population, the correctional environment provides a unique opportunity to reach these individuals and reduce the spread of viral hepatitis. PMID- 16271548 TI - Office-based approach to the implementation of a hepatitis immunization program. AB - To achieve high immunization levels among populations indicated to receive hepatitis A and/or B vaccine, strategies must be implemented to integrate vaccination into routine medical care. Such strategies must address the educational needs and communication skills of providers, patients' awareness of the diseases and the vaccines, the complexity of existing office systems, documentation issues including the use of immunization registries, reimbursement and third-party coverage issues, and ongoing office-based quality assurance. This article describes office-based approaches to implementing hepatitis vaccination programs and some of the tools available to facilitate this process. PMID- 16271549 TI - Expanding the use of hepatitis vaccines in obstetrics and gynecology. AB - Current pediatric vaccination schedules in the United States are based on age. In contrast, adults are vaccinated against most infectious diseases, including hepatitis A and B, only when they are identified as at high risk or after a known exposure. There are multiple risk factors for hepatitis A and B of which clinicians often are not aware. Consequently, many persons at high risk are never vaccinated. Universal vaccination against hepatitis A and B is recommended for adults to halt transmission of the virus and prevent long-term sequelae. Women and their offspring are an especially important population to consider in efforts to reduce the incidence of hepatitis. The following recommendations for expanding the use of hepatitis vaccines in obstetrics and gynecology are made in this article: all older adolescents and adults should be vaccinated regardless of risk factors, and greater efforts should be made to educate physicians about the need to vaccinate their patients against hepatitis. PMID- 16271550 TI - Public health perspective on vaccine-preventable hepatitis: integrating hepatitis A and B vaccines into public health settings. AB - Hepatitis A and B vaccinations can and should be integrated into public health settings that serve adults at high risk for infection (e.g., sexually transmitted disease and/or human immunodeficiency virus clinics, criminal justice settings), and a policy of universal immunization may be the best way to accomplish this goal in these settings. Although hepatitis vaccines should be given to all susceptible persons at risk, many opportunities to vaccinate adults at high risk are missed, and there are several barriers and challenges to vaccination of adults. These challenges and barriers can be overcome. Successful integration of hepatitis vaccination for adults into existing public health services and clinics has been accomplished across the United States at both state and local levels. Additional funds must be provided for the infrastructure and purchase of vaccines for adults in these settings. PMID- 16271551 TI - Conservative correction of leg-length discrepancies of 10mm or less for the relief of chronic low back pain. AB - OBJECTIVE: To study whether conservative correction in a leg-length discrepancy (LLD) of 10mm or less in patients with chronic low back pain (CLBP) can relieve pain. DESIGN: Randomized, controlled intervention study, with a mean follow-up duration of 10 weeks. SETTING: Physical therapy clinic of the national health services. PARTICIPANTS: Thirty-three patients with CLBP were screened for an LLD of 10mm or less, which was measured with ultrasound. Patients were randomly divided into intervention and control groups. INTERVENTION: In 22 patients, LLD was corrected by applying individually fitted shoe inserts. In 11 patients, LLD was not corrected. MAIN OUTCOME MEASURES: Chronic pain intensity (visual analog scale) and disability score (Roland-Morris Disability Questionnaire). RESULTS: Shoe inserts significantly reduced both pain intensity (P<.001) and disability (P<.05). A moderate positive correlation was found between LLD and the degree of pain relief after wearing shoe inserts (r=.47). CONCLUSIONS: Shoe inserts appear to reduce CLBP and functional disability in patients with LLDs of 10mm or less. Shoe inserts are simple, noninvasive, and inexpensive therapeutic means that can be added to the treatment of CLBP. PMID- 16271552 TI - Racial disparities in outcomes of inpatient stroke rehabilitation. AB - OBJECTIVE: To determine whether race is associated with outcomes of inpatient stroke rehabilitation. DESIGN: Retrospective cohort study. SETTING: A community based inpatient rehabilitation facility. PARTICIPANTS: Poststroke patients (N=1002) admitted to a community-based inpatient rehabilitation facility between 1995 and 2001. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Functional improvement at discharge from the rehabilitation facility, discharge disposition, and functional improvement at 3 months after discharge. Inpatient and follow-up data were collected from the facility's electronic patient database. We used the FIM instrument to assess functional status at admission, discharge, and follow up. RESULTS: In multivariable models, blacks achieved less functional improvement at discharge (-1.9 FIM points, P=.02) compared with whites and, despite worse FIM scores, were more likely to be discharged to home (adjusted odds ratio=1.7; 95% confidence interval, 1.1-2.5). Although Asian-American patients did not differ from whites in terms of functional improvement at discharge or disposition, they had less improvement at 3 months following discharge (-6.3 FIM points, P=.005). CONCLUSIONS: We identified racial disparities in poststroke outcomes in a community-based inpatient rehabilitation facility. Future research in stroke rehabilitation should explore the consistency of these findings across settings and if they are confirmed, identify explanatory mediators to better inform efforts to eliminate racial disparities. PMID- 16271553 TI - Beyond function: predicting participation in a rehabilitation cohort. AB - OBJECTIVES: To monitor participation in a rehabilitation cohort and to identify determinants of change during a 12-month period posthospitalization following the onset of one of several major disabling conditions. DESIGN: Cohort study. SETTING: Postacute care rehabilitation settings. PARTICIPANTS: Adults (N=435) aged 18 years and older with complex medical, lower-extremity orthopedic, and major neurologic impairments. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: At 1-, 6-, and 12-month follow-ups, community participation and social and home participation were assessed by personal interviews using the Participation Measure for Post-Acute Care. Information on potential determinants was abstracted from the medical chart and by personal interview using standardized instruments. RESULTS: On average, rehabilitation patients achieved modest improvements in their levels of community participation during the first 6 months after acute hospitalization. In contrast, these same patients displayed a modest loss in social and home participation levels during the follow-up period. Activity limitations were the dominant factors that explained much of the variance in the extent of community participation achieved by patients. Personal and social environmental factors played a major role in predicting levels of social and home participation. CONCLUSIONS: The focus of rehabilitation interventions aimed at achieving posthospital participation requires careful consideration of the specific domain of participation that is being targeted. PMID- 16271555 TI - Performance in different proprioceptive tests does not correlate in ankles with recurrent sprain. AB - OBJECTIVE: To determine the relation between 2 proprioceptive tests, movement detection and movement discrimination, at the ankle. DESIGN: A cross-sectional descriptive study. SETTING: Research laboratory. PARTICIPANTS: Eighteen subjects with recurrent ankle inversion sprain. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Threshold to detection of movement was tested for inversion and eversion movements at 3 velocities (0.1 degrees , 0.5 degrees , 2.5 degrees /s). Movement discrimination was tested for plantarflexion and inversion movements. The tests were performed in random order, and the velocity and movements were randomized within each test paradigm. Correlations (Pearson r) were calculated between movement detection and movement discrimination. RESULTS: Correlation within each proprioceptive paradigm was poor to moderate: for movement detection, correlations among movement directions at each velocity ranged from r equal to .53 to r equal to .54; for movement discrimination correlation was r equal to .49. There was poor correlation between the scores for the 2 tasks in 10 of the 12 comparisons (r range, -.02 to -.36). CONCLUSIONS: These findings show that performance in different proprioceptive tests is not well correlated and, therefore, that general proprioceptive status cannot be inferred from assessment of a single proprioceptive test. PMID- 16271554 TI - The influence of perceived pain on satisfaction with community participation after hospital discharge. AB - OBJECTIVE: To examine the association between pain and satisfaction with community participation for patients with stroke approximately 4 months after discharge from inpatient medical rehabilitation. DESIGN: A cross-sectional study from 2001 to 2002 using information from the IT HealthTrack database. SETTING: Community based. PARTICIPANTS: The study included 1673 patients with stroke aged 40 years or older discharged from inpatient medical rehabilitation. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Primary measures included pain ratings and satisfaction with community participation. RESULTS: The sample was 49.4% female and 81.1% non-Hispanic white. One third of the sample reported mild to severe pain. Pain ratings showed a significant and inverse association with satisfaction with community participation. The association remained significant with adjustment for possible confounding factors and with stratification by right and left body involvement. Each 1-level increase in pain rating score (range, 0-10) was associated with a 12% decreased odds ratio (OR=.88; 95% confidence interval, .84-.93) of being satisfied with community participation after adjustment for age, sex, marital status, ethnicity, and motor and cognitive functional status. CONCLUSIONS: In persons with stroke, high pain rating scores were significantly associated with lower satisfaction with community participation. Effective pain management may increase level of satisfaction with community participation in persons with stroke. PMID- 16271556 TI - A comparison of hydrostatic weighing and air displacement plethysmography in adults with spinal cord injury. AB - OBJECTIVES: To compare (1) total body volume (V(b)) and density (D(b)) measurements obtained by hydrostatic weighing (HW) and air displacement plethysmography (ADP) in adults with spinal cord injury (SCI); (2) measured and predicted thoracic gas volume (V(TG)); and (3) differences in percentage of fat measurements using ADP-obtained D(b) and HW-obtained D(b) measures that were interchanged in a 4-compartment body composition model (4-comp %fat). DESIGN: Twenty adults with SCI underwent ADP and V(TG), and HW testing. In a subgroup (n=13) of subjects, 4-comp %fat procedures were computed. SETTING: Research laboratories in a university setting. PARTICIPANTS: Twenty adults with SCI below the T3 vertebrae and motor complete paraplegia. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Statistical analyses, including determination of group mean differences, shared variance, total error, and 95% confidence intervals. RESULTS: The 2 methods yielded small yet significantly different V(b) and D(b). The groups' mean V(TG) did not differ significantly, but the large relative differences indicated an unacceptable amount of individual error. When the 4-comp %fat measurements were compared, there was a trend toward significant differences (P=.08). CONCLUSIONS: ADP is a valid alternative method of determining the V(b) and D(b) in adults with SCI; however, the predicted V(TG) should be used with caution. PMID- 16271557 TI - Botulinum toxin type A injections for treating neurogenic detrusor overactivity combined with low-compliance bladder in patients with spinal cord lesions. AB - OBJECTIVE: To evaluate botulinum toxin type A (BTX-A) for treating neurogenic detrusor overactivity in patients with spinal cord lesions, including those with abnormally low bladder compliance. DESIGN: Nonrandomized, before-after trial (9 mo follow-up). SETTING: Hospitalized care. PARTICIPANTS: Ten patients with high detrusor contraction pressure and/or poor response to oxyphencyclimine with incontinence selected as a consecutive sample. INTERVENTION: BTX-A (Botox; 300U) was injected into the detrusor muscle. MAIN OUTCOME MEASURES: Urinary continence, functional bladder capacity, bladder compliance, detrusor contraction pressure, and volume at first reflex voiding. Measurements were taken before and 6, 16, and 36 weeks posttreatment. RESULTS: Six weeks after treatment, complete continence was restored in 7 patients without oxyphencyclimine. Mean functional bladder capacity (P=.008), compliance (P=.012), and reflex volume (P=.045) significantly increased, whereas maximal detrusor contraction pressure significantly decreased (P<.001). Urodynamic variables remained significantly improved at 16 weeks, but values were returning toward baseline levels by 36 weeks. The procedure was generally uneventful, without any serious side effects. CONCLUSIONS: BTX-A injections are an effective, well-tolerated treatment for neurogenic detrusor overactivity in patients with spinal cord lesions, even in patients with abnormally low bladder compliance. Patients may require repeat injections after 16 weeks to remain continent. PMID- 16271558 TI - Subject- and injury-related factors influencing the course of manual wheelchair skill performance during initial inpatient rehabilitation of persons with spinal cord injury. AB - OBJECTIVES: To study changes in wheelchair skills in subjects with spinal cord injury (SCI) during rehabilitation; to determine whether changes in wheelchair skill performance are related to the subject, lesion characteristics, secondary complications, and upper extremity pain; and to investigate if wheelchair skill performance at discharge can be predicted from these features. DESIGN: Longitudinal. Subjects performed the Wheelchair Circuit 3 times during rehabilitation: at admission (t1), 3 months later (t2), and at discharge (t3). SETTING: Eight rehabilitation centers in the Netherlands. PARTICIPANTS: One hundred twenty-one subjects with SCI. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: The Wheelchair Circuit consists of 8 wheelchair skills and results in 3 test scores: ability, performance time, and physical strain. RESULTS: All the scores of the Wheelchair Circuit improved significantly between t1 and t2, and between t2 and t3. The scores were related to age and lesion level, whereas changes in scores were related to age, sex, lesion level, and secondary complications. The variables age, body mass index, sex, lesion level, motor completeness, and secondary complications contributed significantly to the prediction of the scores at t3. CONCLUSIONS: Wheelchair skill performance improved during rehabilitation. Personal and lesion characteristics are most important for improving wheelchair skill performance and predicting wheelchair skill performance. PMID- 16271559 TI - Significant spinal injury resulting from low-level accelerations: a case series of roller coaster injuries. AB - OBJECTIVES: To describe a cohort of significantly injured roller coaster riders and the likely levels of acceleration at which the injuries occurred, and to compare these data with contemporary efforts to define a lower limit of acceleration below which no significant spinal injury is likely to occur. DESIGN: A retrospective case series of roller coaster ride-induced significant spinal injuries. SETTING: Injury incident records and emergency medical service records for the Rattler roller coaster in San Antonio, TX, were evaluated for a 19-month period in 1992 and 1993. Medical records for the more significant injuries were also reviewed and the specific injuries were tabulated, along with the demographics of the cohort. PARTICIPANTS: There were 932,000 riders of the Rattler roller coaster, estimated to represent between 300,000 and 600,000 individual riders. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Injury incident reports and medical record review. RESULTS: It is estimated that there were a total of 656 neck and back injuries during the study period, and 39 were considered significant by the study inclusion criteria. Seventy-two percent (28/39) of the injured subjects sustained a cervical disk injury; 71% of these injuries were at C5-6 (15 disk herniations, 5 symptomatic disk bulges) and 54% were at C6-7 (11 disk herniations, 4 symptomatic disk bulges). In the lumbar spine, the most frequent injury was a symptomatic disk bulge (20% of the cohort), followed by vertebral body compression fracture (18%), and L4-5 or L5-S1 disk herniation (13%). Accelerometry testing of passengers and train cars indicated a peak of 4.5 to 5g of vertical or axial acceleration and 1.5g of lateral acceleration over approximately 100ms (0.1s) on both. CONCLUSIONS: The results of this study suggest that there is no established minimum threshold of significant spine injury. The greatest explanation for injury from traumatic loading of the spine is individual susceptibility to injury, an unpredictable variable. PMID- 16271560 TI - Exploiting interlimb coupling to improve paretic arm reaching performance in people with chronic stroke. AB - OBJECTIVE: To determine whether paretic arm reaching performance is improved in bilateral compared with unilateral conditions. DESIGN: Cohort study. SETTING: University human performance laboratory. PARTICIPANTS: Thirty-two subjects with chronic stroke (57+/-14y; on Fugl-Meyer Assessment arm score, 37+/-14). INTERVENTION: Unilateral and bilateral reaching. Bilateral tasks included varying levels of weight on the nonparetic hand. MAIN OUTCOME MEASURES: An electromagnetic tracking system recorded hand peak acceleration, velocity, and movement time. A 2-way repeated-measures analysis of variance and Tukey-adjusted pairwise comparisons were used to analyze the results (alpha=.05). RESULTS: Paretic differed significantly from nonparetic peak acceleration and velocity in unilateral reaching but not bilateral reaching. Within limbs, the paretic arm attained a higher peak acceleration (P<.001) and velocity (P=.03) in the bilateral compared with the unilateral task, but movement time was unchanged between tasks. Nonparetic peak acceleration was higher (P=.015), velocity was unchanged, and movement time increased (P=.005) in the bilateral compared with the unilateral task. The addition of a weight to the nonparetic arm during bilateral reaching did not result in further improvement in paretic arm performance. CONCLUSIONS: Interlimb coupling effects during bilateral reaching are retained even after chronic stroke and can be used to produce an immediate improvement in paretic arm reaching performance. PMID- 16271561 TI - Clinical characteristics and rehabilitation outcomes of patients with posterior cerebral artery stroke. AB - OBJECTIVES: To describe the demographics, clinical profile, and functional outcomes in posterior cerebral artery (PCA) stroke and to identify factors associated with functional change during rehabilitation and discharge disposition. DESIGN: Retrospective study of patients with PCA stroke admitted to a rehabilitation hospital over an 8-year period. SETTING: Free-standing urban rehabilitation hospital in the United States. PARTICIPANTS: Eighty-nine consecutive patients with PCA stroke (48 men, 41 women; mean age, 71.5y) met inclusion criteria. INTERVENTION: Inpatient multidisciplinary comprehensive rehabilitation program. MAIN OUTCOME MEASURES: Demographic, clinical, and discharge disposition information were collected. Functional status was measured using the FIM instrument, recorded at admission and discharge. The main outcome measures were the discharge total FIM score, the change in total FIM score (DeltaFIM), and the discharge disposition. Multiple and logistic regression analyses were performed to identify factors associated with the main outcome measures. RESULTS: The most common impairments were motor paresis (65%), followed by visual field defects (54%) and confusion or agitation (43%). The mean discharge total FIM score +/- standard deviation was 88.3+/-28.2. The mean DeltaFIM was 23.3+/-16.4. Fifty-five (62%) patients were discharged home. On multiple regression analysis, higher admission total FIM score, longer length of stay (LOS), and a rehabilitation stay free of interruptions were associated with higher discharge total FIM score and greater DeltaFIM. Absence of diabetes mellitus and younger age were also associated with higher discharge total FIM scores, and male sex had greater DeltaFIM. On logistic regression analysis, younger patients, higher discharge FIM scores, presence of a caregiver, and the nonnecessity for 24-hour support were associated with a discharge to home. CONCLUSIONS: Motor, visual, and cognitive impairments are common in PCA stroke, and good functional gains are achievable after comprehensive rehabilitation. Higher admission FIM scores, longer LOS, and younger and male patients were associated with better functional outcomes. Most patients were discharged home, particularly those with caregivers and those for whom 24-hour support was not required. Further research should aim at the development of functional outcome measures of greater breadth and sensitivity to visual and cognitive deficits and should compare PCA stroke outcomes with outcomes of strokes in other vascular territories. PMID- 16271562 TI - Submental surface electromyographic measurement and pharyngeal pressures during normal and effortful swallowing. AB - OBJECTIVE: To evaluate the influence of 2 swallowing maneuvers on anterior suprahyoid surface electromyographic measurement and pharyngeal manometric pressure. DESIGN: Correlational analysis of biomechanic measures of swallowing. SETTING: Research laboratory in a community hospital. PARTICIPANTS: A consecutive volunteer sample of 22 healthy subjects (mean, 29.7y). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Peak amplitude of submental surface electromyographic and pharyngeal manometric pressure at 3 locations. RESULTS: Effortful swallow generated greater pharyngeal pressure than normal swallow at the 2 proximal pharyngeal sensors (sensor 1: P=.017; sensor 2: P=.009) and lower pressure at the distal sensor (upper esophageal sphincter) (P<.001). Pressure in the upper pharynx was lower than that in the lower pharynx (P=.027). Effortful swallow generated greater surface electromyographic amplitudes than normal swallowing (P<.001). A statistically significant but weak negative correlation was identified between surface electromyographic and mid-pharyngeal pressure for normal swallowing condition (r=-.21, P<.01). For the effortful swallowing condition, statistically significant but weak negative correlations were identified between surface electromyographic and pressure measurements at all sensors (sensor 1: r=-.16, P=.02; sensor 2: r=-.30, P<.01; sensor 3: r=-.18, P<.01). CONCLUSIONS: There is a significant change in both suprahyoid surface electromyographic and pharyngeal pressures during effortful swallow compared with normal swallow. PMID- 16271563 TI - Impact of a home-based activity and dietary intervention in people with slowly progressive neuromuscular diseases. AB - OBJECTIVE: To determine whether a home-based activity and dietary intervention can increase activity level, reduce caloric intake, and impact positively components of metabolic syndrome in a disabled population. DESIGN: Testing occurred at 3 points during the 6-month intervention period (baseline, 3mo, 6mo) and at 6 months postintervention. Each test point included laboratory testing of anthropometric and metabolic variables and 3 days of home-based activity and dietary monitoring. A personally tailored activity and dietary prescription based on baseline testing was implemented during the 6-month intervention period. SETTING: Human performance laboratory of a university and each subject's home. PARTICIPANTS: Twenty adult volunteer ambulatory subjects with several types of slowly progressive neuromuscular disease (NMD). INTERVENTION: Using a pedometer, subjects were instructed to increase number of steps by 25% over their baseline determined from home monitoring. An individualized dietary prescription was provided focusing on problematic issues identified from the baseline dietary profile. MAIN OUTCOME MEASURES: Body composition, physical activity, dietary intake, energy expenditure, gait efficiency, metabolic variables, and quality of life. RESULTS: At the end of the protocol, mean step count increased approximately 27% above baseline (P=.001) and caloric intake decreased over 300kcal/d (P=.002). Body fat percentage significantly decreased (from 33.3%+/ 1.5% to 32.6%+/-1.6%, P=.032). Gait efficiency did not change, and metabolic variables did not show statistically significant improvement, although 2 of the 5 subjects originally meeting the criteria for metabolic syndrome at baseline no longer met the criteria at the end of the intervention period. Six months after completing the protocol, caloric intake remained significantly reduced (P=.02), but although mean step count remained elevated, it was not statistically significant. CONCLUSIONS: Using a home-based protocol, people with NMD can increase activity and reduce caloric intake. Although this 6-month program showed positive changes, it was insufficient to affect risk factors associated with metabolic syndrome. It remains to be seen if a program longer than 6 months or a more rigorous program could lead to a reduction in the risk factors associated with metabolic syndrome. PMID- 16271564 TI - Strategies that improve paralyzed human quadriceps femoris muscle performance during repetitive, nonisometric contractions. AB - OBJECTIVE: To determine the effect of combining different stimulation frequencies on the ability of paralyzed human quadriceps muscle to produce a 50 degrees knee excursion repetitively when starting at 90 degrees of flexion. DESIGN: Repeated measures design. SETTING: Clinical research laboratory. PARTICIPANTS: Complete data were collected from 9 subjects aged 11 to 25 years (mean +/- standard deviation, 17.1+/-4.5y) with spinal cord injury (SCI). INTERVENTION: Three protocols were each tested during separate sessions: 20-Hz trains of pulses followed by 66-Hz trains (C20+66), 33-Hz trains followed by 66-Hz trains (C33+66), and 66-Hz trains alone (C66). For each frequency, stimulation was repeated until the knee failed to produce a 50 degrees excursion. This approach allowed us to evaluate the response to stimulation with 20-, 33-, and 66-Hz and combinations of 20- and 66-Hz and 33- and 66-Hz trains. MAIN OUTCOME MEASURE: Number of successful contractions. RESULTS: The C20 and C33 did not differ (mean, 41.0+/-12.6 excursions and 42.0+/-12.3 excursions, respectively), and each produced more excursions than the C66 protocol. The C20+66 and C33+66 protocols produced 51.4+/-15.0 and 44.9+/-13.6 excursions, respectively, and the C20+66 was the best protocol overall (all Por=2.5) (P=.004). CONCLUSIONS: The differences in COP-COM distances between these H&Y groups suggest that patients with PD who have impaired postural control produce shorter COM-COP distances than do persons without clinically detectable balance impairment. This method of evaluation could prove a useful quantitative index to examine the impact of interventions designed to improve ambulation and balance in PD. PMID- 16271568 TI - Functional measures across neurologic disease states: analysis of factors in common. AB - OBJECTIVE: To describe the underlying dimensions for a range of functional measures across 3 neurologic diseases at different time points. DESIGN: Multiple cohort study. SETTING: Combination of public hospital wards and community. PARTICIPANTS: Patients (N=308) from 3 cohorts: paralytic poliomyelitis (n=38), mean of 25 years previously, assessed once; acute stroke admitted to hospital and followed up for 12 months postdischarge (n=181); and traumatic brain injury (TBI), admitted to hospital and followed up for 12 months postdischarge (n=89). INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: The Barthel Index, FIM instrument, Medical Outcomes Study 36-Item Short-Form Health Survey (SF-36) physical component score (PCS) and mental component score (MCS), Community Integration Questionnaire (CIQ), and the London Handicap Score (LHS). Measures were compared at 2 time points: pre-event status for stroke, TBI and the polio cohort, and 12-month postdischarge status for stroke and TBI. RESULTS: The different measures generally correlated highly within disease states at each time point. Principal components analysis revealed 2 underlying dimensions, a physical dimension onto which loaded the Barthel Index, FIM, PCS, and LHS and a cognitive/emotional dimension onto which loaded the MCS and, for subjects with polio, the CIQ. These 2 dimensions accounted for 69% of the variance in measures at the pre-event time point and 85% of the variance at the 12-month time point. CONCLUSIONS: These data suggest 2 basic underlying dimensions across a wide range of measures in 3 different neurologic conditions even at different time points. Most of the variation in the measures can be captured using the 2 component summary scores (PCS, MCS) of the SF-36. PMID- 16271567 TI - The effect of walking speed on lower-extremity joint powers among elderly adults who exhibit low physical performance. AB - OBJECTIVES: To compare peak joint powers and joint angles between comfortable and fast walking speeds among a group of elderly adults who exhibit low physical performance, and to test the primary hypothesis that peak ankle powers would not change when walking speed was increased, but that peak hip power output would increase significantly with speed. DESIGN: Three-dimensional analysis of joint kinematics and kinetics during comfortable and fast walking by both healthy and low-performing elderly adults (age, >70y). SETTING: Gait laboratory. PARTICIPANTS: Twenty-four healthy elderly adults and 27 elders who exhibited low performance on a standard battery of walking, standing balance, and chair-rise tasks that places them at risk of mobility-related disability. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Peak lower-extremity joint powers and joint angles. RESULTS: Low-performing elders increased both ankle and hip power outputs to increase walking speed. However, peak ankle power remained significantly below that of the healthy elderly adults even when the low-performing elders walked at a faster gait speed. Joint-power changes in the low-performing elderly were accompanied by a reduction in hip extension and ankle dorsiflexion, and an increase in transverse pelvic rotation. CONCLUSIONS: Compared with healthy elderly, the low-performing elderly adults showed speed-independent differences in ankle and hip mechanics that may reflect underlying neuromuscular impairments. In particular, an understanding of the interdependent contributions of hip flexibility and ankle power limitations seem important to inform interventions to maintain gait into advanced age. PMID- 16271569 TI - Center of mass movement and energy transfer during walking in children with cerebral palsy. AB - OBJECTIVE: To gain insight into the mechanical inefficiencies of gait patterns used by children with spastic diplegia by analysis of center of mass (COM) movement and energy recovery. DESIGN: Prospective study using between-group measures to analyze differences between children with cerebral palsy (CP) and age matched controls without CP. SETTING: Assessments were performed in a gait laboratory. PARTICIPANTS: Fifteen children with spastic diplegia and 6 age matched controls without CP with a mean age of 9.7 years. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: Gait data assessed included temporal-distance factors, COM vertical excursion, work done on the COM, and the percentage of energy transferred and relative phase between the potential and kinetic energy. RESULTS: Children with CP had a 33% smaller energy recovery factor than the controls (P<.001). They also had 60% greater COM vertical excursion (P<.02) and a poorer phasic relation between potential and kinetic energies (P<.02), both of which contributed to greater mechanical work performed (P<.003). CONCLUSIONS: Compared with the age-matched controls without CP, the children with CP were mechanically less efficient in their gait. Interventions that promote heel contact and roll over and greater knee stability to better utilize the kinetic energy of push-off could improve walking efficiency. PMID- 16271570 TI - Efficacy of forced-use therapy in hemiplegic cerebral palsy. AB - OBJECTIVE: To determine the efficacy of forced-use therapy (FUT) on the improvement of upper-extremity function in children with hemiplegic cerebral palsy (CP). DESIGN: Prospective case series. SETTING: Outpatient ambulatory clinic in South Korea. PARTICIPANTS: Thirty-one patients with hemiplegic CP were assigned to the FUT group (n=18) or to the control group (n=13). The mean age of the patients in the FUT group was 33.2 months and in the control group it was 43.2 months. INTERVENTIONS: The FUT group wore a short-arm Scotchcast on the unaffected arm for 6 weeks and also participated in a conventional rehabilitation program that included stretching exercises and functional occupational therapy for the upper extremity. The control group underwent the conventional rehabilitation program only. MAIN OUTCOME MEASURE: Hand function tests, including the box and block test (BBT), Erhardt Developmental Prehension Assessment (EDPA), and WeeFIM instrument taken before and after 6 weeks of treatment. RESULTS: Before treatment, there was no significant difference between groups in the BBT, EDPA, and WeeFIM scores. After 6 weeks of treatment, however, the FUT group showed significant improvement in the affected arm in the BBT and EDPA scores, compared with the control group (P<.05). The self-care score on the WeeFIM was also significantly improved in the FUT group (P<.05). CONCLUSIONS: FUT combined with a conventional rehabilitation program appears to be more effective than a rehabilitation program alone in improving affected hand function in children with hemiplegic CP. PMID- 16271571 TI - Predictive model for congenital muscular torticollis: analysis of 1021 infants with sonography. AB - OBJECTIVE: To construct a predictive model to foretell congenital muscular torticollis (CMT) on the basis of clinical correlates. DESIGN: Correlation study. SETTING: Regional hospital. PARTICIPANTS: A consecutive series of 1021 newborn infants. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURE: Participants underwent portable ultrasonography to diagnose CMT. Significant clinical correlates were identified to construct a predictive model using the logistic regression model. RESULTS: Forty of 1021 infants were diagnosed with CMT using ultrasonography, yielding an overall incidence of 3.92%. Birth body length (odds ratio [OR]=1.38; 95% confidence interval [CI], 1.49-2.38), facial asymmetry (OR=21.75; 95% CI, 6.6-71.7), plagiocephaly (OR=22.3; 95% CI, 7.01-70.95), perineal trauma during delivery (OR=4.26; 95% CI, 1.25-14.52), and primiparity (OR=6.32; 95% CI, 2.34-17.04) were significant correlates. A predictive logistic regression model with the incorporation of these 4 correlates was developed. We used cross-validation with a receiver operating characteristic curve to validate the predictive model. CONCLUSIONS: Our study successfully developed a quantitative predictive model for estimating the risk of CMT on the basis of clinical correlates only. This model has good discriminative ability for classifying CMT and non-CMT by yielding acceptable values of false-negative and false-positive cases. PMID- 16271572 TI - Improvement of a questionnaire measuring activity limitations in rising and sitting down in patients with lower-extremity disorders living at home. AB - OBJECTIVE: To improve a self-administered questionnaire that includes 42 dichotomous items and measures activity limitations in rising and sitting down (R&S) in patients with lower-extremity disorders who live at home. DESIGN: Cross sectional study. SETTING: Outpatient clinics of secondary and tertiary care centers. PARTICIPANTS: Patients (N=759; 47% men; mean age +/- standard deviation, 60.7+/-15.2y) living at home, with lower-extremity disorders resulting from stroke, poliomyelitis, osteoarthritis, amputation, and complex regional pain syndrome type I. INTERVENTIONS: Not applicable. MAIN OUTCOME MEASURES: (1) Unidimensionality, indicating that items assess only a single construct; (2) fit with the one-parameter logistic model (OPLM), yielding information about patient and item location parameters; (3) intratest reliability, indicating consistency of patients' item scores; and (4) content validity, indicating completeness with which the items cover the important aspects of the construct that they are attempting to represent. RESULTS: Thirty-nine of 42 items: (1) loaded on 1 component (variance explained, 59%; item component loadings, >or=.51), (2) showed good fit with the OPLM (P=.15), (3) had a good intratest reliability (Cronbach alpha=.96), and (4) had a good content validity (all important aspects represented). CONCLUSIONS: A unidimensional scale that fits with the OPLM has been developed for measuring activity limitations in R&S in patients with lower extremity disorders who live at home. PMID- 16271573 TI - Fitting a bilateral transhumeral amputee with utensil prostheses and their functional assessment 10 years later: a case report. AB - Traumatic transhumeral amputations resulting from electric burn injury are uncommon and present a significant rehabilitation challenge. Compensating for loss of fine, coordinated function of the upper extremities with prostheses is difficult medically, technologically, psychologically, and socially. We followed up a patient with traumatic bilateral transhumeral amputation who was fitted with specially designed bilateral low-temperature utensil prostheses for 10 years. A bilateral utensil prosthesis consists of 2 thermoplastic sockets, an elastic harness, 2 utensil holders, and several different utensils. The characteristics of utensil prostheses are low cost, quick fabrication, and responsiveness to a patient's needs. Ten years after the patient's first prosthesis fitting, he still used these specially designed prostheses. By using these devices and his feet, the patient has regained independence in most activities of daily living and gained a new working skill. PMID- 16271574 TI - Cervical radiculopathy caused by neural foraminal migration of a herniated calcified intervertebral disk in childhood: a case report. AB - Childhood intervertebral disk calcification is a rare clinical entity. Although its clinical course is usually benign, nerve root irritation or spinal cord compression can occasionally occur. We present the clinical and radiologic findings of a 9-year-old boy with cervical radiculopathy due to a herniated calcified intervertebral disk, which developed suddenly after swimming for 1 hour. Radiologic findings indicated that a calcified nucleus pulposus at the C6-7 level herniated into the spinal canal and migrated far into the right C6-7 neural foramen. Surgical management was performed 8 weeks after the onset of symptoms, because the initial presenting symptoms persisted despite conservative treatment. In children, calcified intervertebral disks can cause cervical radiculopathy that requires surgical management when they herniate and migrate far into the neural foramen. PMID- 16271575 TI - Cortical reorganization and associated functional motor recovery after virtual reality in patients with chronic stroke: an experimenter-blind preliminary study. AB - OBJECTIVE: To investigate the effects of virtual reality (VR) on cortical reorganization and motor recovery. DESIGN: Nonparametric pre- and posttest design with experimenter blinded. SETTING: University medical center. PARTICIPANTS: Five patients with hemiparesis (age, 59.8+/-3.4y) were recruited. INTERVENTION: Five patients received VR for 60 minutes a day, 5 times a week for 4 weeks. VR was designed to provide a virtual rehabilitation scene where the intensity of practice and sensory feedback could be systematically manipulated to provide the most appropriate, individualized motor retraining program. MAIN OUTCOME MEASURES: Cortical activation and associated motor recovery were measured before and after VR using functional magnetic resonance imaging and standardized motor tests, respectively. Nonparametric tests were used at P less than .05. RESULTS: Prior to VR, the bilateral primary sensorimotor cortices (SM1s), contralesional premotor cortex, and contralesional or ipsilesional supplementary motor area were activated. After VR, the altered activations disappeared and predominantly the ipsilesional SM1 was activated (P<.05). Motor function was improved (P<.05). CONCLUSIONS: This is a novel demonstration of VR-induced neuroplastic changes and associated motor recovery in chronic stroke. PMID- 16271576 TI - Electromyographic evaluation of cervical radiculopathy. PMID- 16271578 TI - Use of the Berg Balance Scale in rehabilitation evaluation of patients with Parkinson's disease. PMID- 16271580 TI - Classifying functional ambulation. PMID- 16271581 TI - The "perfect storm" for minorities with arthritis. PMID- 16271584 TI - Analysis of the transient expansion behavior and design optimization of coronary stents by finite element method. AB - The percutaneous transluminal coronary angioplasty (PTCA) assisted with stenting technique has become a primary therapy to coronary heart disease. In practice, the structure conditions of both ends of stent/balloon system influence a stent's instantaneous expansion behavior. The transitory nonuniform expansion, the so called dogboning, of stent/balloon system is one of the main reasons to induce the acute vascular injury at the two edges of a stent. This kind of vascular injury has a close relationship with the in-stent restenosis. In the present paper, the finite element method (FEM) was applied to simulate the transient expansion process of stent/balloon system with different stent structure and balloon length under the internal pressure. And two types of stent and six collocations of stent and balloon were modeled. Modeling results showed that the dogboning phenomenon can be eliminated by improving geometry of a stent or/and varying the length of balloon over stent. The above modeled results were further confirmed by following in situ observation. PMID- 16271583 TI - Biomechanics of knee ligaments: injury, healing, and repair. AB - Knee ligament injuries are common, particularly in sports and sports related activities. Rupture of these ligaments upsets the balance between knee mobility and stability, resulting in abnormal knee kinematics and damage to other tissues in and around the joint that lead to morbidity and pain. During the past three decades, significant advances have been made in characterizing the biomechanical and biochemical properties of knee ligaments as an individual component as well as their contribution to joint function. Further, significant knowledge on the healing process and replacement of ligaments after rupture have helped to evaluate the effectiveness of various treatment procedures. This review paper provides an overview of the current biological and biomechanical knowledge on normal knee ligaments, as well as ligament healing and reconstruction following injury. Further, it deals with new and exciting functional tissue engineering approaches (ex. growth factors, gene transfer and gene therapy, cell therapy, mechanical factors, and the use of scaffolding materials) aimed at improving the healing of ligaments as well as the interface between a replacement graft and bone. In addition, it explores the anatomical, biological and functional perspectives of current reconstruction procedures. Through the utilization of robotics technology and computational modeling, there is a better understanding of the kinematics of the knee and the in situ forces in knee ligaments and replacement grafts. The research summarized here is multidisciplinary and cutting edge that will ultimately help improve the treatment of ligament injuries. The material presented should serve as an inspiration to future investigators. PMID- 16271585 TI - Bone temperature estimation during orthopaedic round bur milling operations. AB - Heat generation during orthopaedic bone cutting operations may cause thermal bone damage. During the bone cutting, the maximum temperature occurs at the point of contact between the bone and the cutting tool. However, this temperature is difficult to measure. Many researchers have attempted to measure this temperature using a thermocouple; however, limitations of the thermocouple makes it difficult to determine the maximum temperature at the point of contact. In order to solve this problem, in this study, two infrared thermometers are used to measure the fresh-milled surface temperature, and the maximum temperature was extrapolated by a moving plane heat source solution. The estimated maximum temperature increment varied from 49 to 115 degrees C under various cutting conditions. These results showed that the thermal damage may reach up to 1.9 mm in depth during round bur milling. A larger feed rate and a smaller cutting depth decreased the maximum temperature. PMID- 16271586 TI - Viscoelastic finite element analysis of an all-ceramic fixed partial denture. AB - In recent years metal-free ceramic systems have become increasingly popular in dental practice because of their superior aesthetics, chemical durability and biocompatibility. Recently, manufacturers have proposed new dental ceramic systems that are advertised as being suitable for posterior fixed partial dentures (FPDs). Reports indicate that some of these systems have exhibited poor clinical performance. The objective of this study was to use the viscoelastic option of the ANSYS finite element program to calculate residual stresses in an all-ceramic FPD for four ceramic-ceramic combinations. A three-dimensional finite element model of the FPD was constructed from digitized scanning data and calculations were performed for four systems: (1) IPS Empress 2, a glass veneering material, and Empress 2 core ceramic; (2) IPS Eris a low fusing fluorapatite-containing glass-veneering ceramic, and Empress 2 core ceramic; (3) IPS Empress 2 veneer and an experimental lithium-disilicate-based core ceramic; and (4) IPS Eris and an experimental lithium-disilicate-based core ceramic. The maximum residual tensile stresses in the veneer layer for these combinations are as follows: (1) 77 MPa, (2) 108 MPa, (3) 79 MPa, and (4) 100 MPa. These stresses are relatively high compared to the flexural strengths of these materials. In all cases, the maximum residual tensile stresses in the core frameworks were well below the flexural strengths of these materials. We conclude that the high residual tensile stresses in all-ceramic FPDs with a layering ceramic may place these systems in jeopardy of failure under occlusal loading in the oral cavity. PMID- 16271587 TI - Viscoelastic behaviour and failure of bovine cancellous bone under constant strain rate. AB - A programme has been established to characterize the long-term behaviour of cancellous bone. Fresh bovine cancellous specimens of dimensions 10 x 10 x 10 mm3 and 10 x 40 x 3.6 mm3 were manufactured and used within the testing programme. Results published in the literature indicate that the long-term behaviour of cancellous bone is well described by a power law, which is a very similar response of typical polymers. So far, dynamic mechanical tests (DMA) in three point bending, under frequencies between 0.01 and 100 Hz at room temperature, confirmed the published results in a qualitative way. Nevertheless, the measured dimensionless damping, tan delta, was slightly higher than the values reported in the literature for the compact bone. The relaxation curves were obtained from dynamic tests and confirmed that bone relaxation modulus can be described by a power law function of time. Tests under constant compression strain rate were performed at four different strain rates: 0.15/s, 0.015/s, 0.0015/s and 0.00015/s and strain rate dependent behaviour was observed. An average elastic bending modulus of 300 MPa was obtained. PMID- 16271588 TI - Tendon healing in interleukin-4 and interleukin-6 knockout mice. AB - Cytokines have been shown to play an important role in tendon and ligament healing by regulating cellular differentiation and activity. The majority of studies that have investigated the role of cytokines in tendon and ligament healing have added them to injured tissue and assessed their effect. Because the efficacy of exogenously applying cytokines is dependent upon many factors such as the correct dosage, timing, and frequency, conflicting results are often reported. To avoid these factors, this study used transgenic mice with knockouts of interleukin-4 (IL4 -/-) and interleukin-6 (IL6 -/-) to investigate their role in tendon healing. Because of the reported roles of both of these cytokines in inflammation and fibroplasia, it was hypothesized that the order of organizational, geometric, and mechanical properties would be (greatest to least) injured IL6 -/-, injured control, and injured IL4 -/- mice. In addition, it was hypothesized that specific cytokines would be upregulated in each knockout group, but not compensate for the lack of IL-4 or IL-6. Mechanical and organizational properties of injured tendons from IL6 -/- mice were inferior to that of control and IL4 -/- mice despite the upregulation of the pro-inflammatory cytokine TNF alpha. Temporal levels of IL-10 and IL-13 in the IL4 -/- mice resulted in comparable and even superior properties when compared to CTL mice. This study shows that IL-6 could not be compensated for and plays an important role in tendon healing. This study also supports the use of this animal model to further investigate tendon healing. PMID- 16271589 TI - Influence of a downstream narrowing on the flow profile in a tube. AB - The distance over which the upstream flow conditions in a tube are disturbed by a stenosis downstream, i.e. the outlet length, was investigated for Reynolds numbers in the range 210-2900. Two methods were used, the Navier-Stokes equations were solved with a computer and a physical model was constructed and maximal velocities were measured with an ultrasound Doppler system. The computer model showed that Re number does not influence the outlet length, varying the stenosis area from 25% to 90% has an effect. However, the outlet length remained small, below 70% of the diameter of the tube. The physical model confirmed for a 75% stenosis that the outlet length is small, this method set the limit at not more than 1.2 times the tube diameter. PMID- 16271590 TI - Determination of the Poisson's ratio of the cell: recovery properties of chondrocytes after release from complete micropipette aspiration. AB - Chondrocytes in articular cartilage are regularly subjected to compression and recovery due to dynamic loading of the joint. Previous studies have investigated the elastic and viscoelastic properties of chondrocytes using micropipette aspiration techniques, but in order to calculate cell properties, these studies have generally assumed that cells are incompressible with a Poisson's ratio of 0.5. The goal of this study was to measure the Poisson's ratio and recovery properties of the chondrocyte by combining theoretical modeling with experimental measures of complete cellular aspiration and release from a micropipette. Chondrocytes isolated from non-osteoarthritic and osteoarthritic cartilage were fully aspirated into a micropipette and allowed to reach mechanical equilibrium. Cells were then extruded from the micropipette and cell volume and morphology were measured throughout the experiment. This experimental procedure was simulated with finite element analysis, modeling the chondrocyte as either a compressible two-mode viscoelastic solid, or as a biphasic viscoelastic material. By fitting the experimental data to the theoretically predicted cell response, the Poisson's ratio and the viscoelastic recovery properties of the cell were determined. The Poisson's ratio of chondrocytes was found to be 0.38 for non osteoarthritic cartilage and 0.36 for osteoarthritic chondrocytes (no significant difference). Osteoarthritic chondrocytes showed an increased recovery time following full aspiration. In contrast to previous assumptions, these findings suggest that chondrocytes are compressible, consistent with previous studies showing cell volume changes with compression of the extracellular matrix. PMID- 16271592 TI - Transverse elasticity and blood perfusion of sciatic nerves under in situ circular compression. AB - Biomechanical properties and microcirculation of peripheral nerves under circular compression are vital factors for nerve repair and for developing neural prostheses. Quasi-static circular compression experiments on six rabbit sciatic nerves were performed. The mean estimated Young's modulus of the sciatic nerves in the transverse direction was 66.9+/-8.0 kPa. The blood perfusion of the nerve started to decrease at a mean pressure of 30.5 mmHg and reached a stable lower level of 30% of pre-compression value at 102.8 mmHg. The findings may make a contribution to safer design of cuff electrodes to be used in neural prostheses. PMID- 16271591 TI - The effects of cellular contraction on aortic valve leaflet flexural stiffness. AB - The aortic valve (AV) leaflet contains a heterogeneous interstitial cell population composed predominantly of myofibroblasts, which contain both fibroblast and smooth muscle cell characteristics. The focus of the present study was to examine aortic valve interstitial cell (AVIC) contractile behavior within the intact leaflet tissue. Circumferential strips of porcine AV leaflets were mechanically tested under flexure, with the AVIC maintained in the normal, contracted, and contraction-inhibited states. Leaflets were flexed both with (WC) and against (AC) the natural leaflet curvature, both before and after the addition of 90 mM KCl to elicit cellular contraction. In addition, a natural basal tonus was also demonstrated by treating the leaflets with 10 microM thapsigargin to completely inhibit AVIC contraction. Results revealed a 48% increase in leaflet stiffness with AVIC contraction (from 703 to 1040 kPa, respectively) when bent in the AC direction (p=0.004), while the WC direction resulted only in 5% increase (from 491 to 516.5 kPa, respectively--not significant) in leaflet stiffness in the active state. Also, the loss of basal tonus of the AVIC population with thapsigargin treatment resulted in 76% (AC, p=0.001) and 54% (WC, p=0.036) decreases in leaflet stiffness at 5 mM KCl levels, while preventing contraction with the addition of 90 mM KCl as expected. We speculate that the observed layer dependent effects of AVIC contraction are primarily due to varying ECM mechanical properties in the ventricularis and fibrosa layers. Moreover, while we have demonstrated that AVIC contractile ability is a significant contributor to AV leaflet bending stiffness, it most likely serves a role in maintaining AV leaflet tissue homeostasis that has yet to be elucidated. PMID- 16271593 TI - Frictional properties of regenerated cartilage in vitro. AB - Although tribological function is the most important mechanical property of articular cartilage, few studies have examined this function in tissue-engineered cartilage. We investigated changes in the frictional properties of cartilage regenerated from the inoculation of rabbit chondrocytes into fibroin sponge. A reciprocating friction-testing apparatus was used to measure the friction coefficient of the regenerated cartilage under a small load. The specimen was slid against a stainless steel plate in a water vessel filled with physiological saline. The applied load was 0.03 N, the stroke length was 20 mm, and the mean sliding velocity was 0.8 mm/s. The friction coefficient of the regenerated cartilage decreased with increasing cultivation time, because a hydrophilic layer of synthesized extracellular matrix was formed on the fibroin sponge surface. The friction coefficient of the regenerated cartilage was as low as that of natural cartilage in the early stages of the sliding tests, but it increased with increasing duration of sliding owing to exudation of interstitial water from the surface layer. PMID- 16271594 TI - Validity of the Pedar Mobile system for vertical force measurement during a seven hour period. AB - Objective measurement of weight bearing during a long-term period can give insight into the postoperative loading of the lower extremity of orthopedic patients to avoid complications. This study investigated the validity of vertical ground reaction force measurements during a long-term period using the Pedar Mobile insole pressure system, by comparing it with a Kistler force platform. In addition, the validity of a new sensor drift correction algorithm to correct for offset drift in the Pedar signal was evaluated. Ground reaction force data were collected during dynamic and static conditions from five healthy subjects every hour for 7 h. A mean offset drift of 14.6% was found after 7 h. After applying the drift correction algorithm the Pedar system showed a high accuracy for the second peak in the ground reaction force-time curve (1.1 to 3.4% difference, p>0.05) and step duration (-2.0 to 4.4% difference, p>0.05). Less accuracy was found for the first peak in the ground reaction force-time curve (5.2 to 12.0% difference; p<0.05 for the first 3 h, p>0.05 for the last 4 h) and, consequently, in the vertical force impulse (5.5 to 11.0% difference, p>0.05). The Pedar Mobile system appeared to be a valid instrument to measure the vertical force during a long-term period when using the drift correction program described in this study. PMID- 16271595 TI - The influence of soft tissue movement on ground reaction forces, joint torques and joint reaction forces in drop landings. AB - The aim of this study was to determine the effects that soft tissue motion has on ground reaction forces, joint torques and joint reaction forces in drop landings. To this end a four body-segment wobbling mass model was developed to reproduce the vertical ground reaction force curve for the first 100 ms of landing. Particular attention was paid to the passive impact phase, while selecting most model parameters a priori, thus permitting examination of the rigid body assumption on system kinetics. A two-dimensional wobbling mass model was developed in DADS (version 9.00, CADSI) to simulate landing from a drop of 43 cm. Subject-specific inertia parameters were calculated for both the rigid links and the wobbling masses. The magnitude and frequency response of the soft tissue of the subject to impulsive loading was measured and used as a criterion for assessing the wobbling mass motion. The model successfully reproduced the vertical ground reaction force for the first 100 ms of the landing with a peak vertical ground reaction force error of 1.2% and root mean square errors of 5% for the first 15 ms and 12% for the first 40 ms. The resultant joint forces and torques were lower for the wobbling mass model compared with a rigid body model, up to nearly 50% lower, indicating the important contribution of the wobbling masses on reducing system loading. PMID- 16271596 TI - Evaluation of a new algorithm to determine the hip joint center. AB - Accurately locating the hip joint center is a challenging and important step in many biomechanical investigations. The purpose of this study was to test the accuracy and robustness of a "pivoting" algorithm used to locate the hip center. We tested the performance of this algorithm with data acquired by manipulating a ball and socket model of the hip through several motion patterns. The smallest mean errors of 2.2+/-0.2 mm occurred with a circumduction motion pattern, while the largest errors of 4.2+/-1.3 mm occurred with single-plane motion (e.g., flexion/extension). Introducing random noise with an amplitude of 30 mm increased the errors by only 1.3+/-0.5 mm with a circumduction motion pattern. The pivoting algorithm performs well in the laboratory, and further work is warranted to evaluate its performance in a clinical setting. PMID- 16271597 TI - Anisotropic hydraulic permeability in compressed articular cartilage. AB - The extent to which articular cartilage hydraulic permeability is anisotropic is largely unknown, despite its importance for understanding mechanisms of joint lubrication, load bearing, transport phenomena, and mechanotransduction. We developed and applied new techniques for the direct measurement of hydraulic permeability within statically compressed adult bovine cartilage explant disks, dissected such that disk axes were perpendicular to the articular surface. Applied pressure gradients were kept small to minimize flow-induced matrix compaction, and fluid outflows were measured by observation of a meniscus in a glass capillary under a microscope. Explant disk geometry under radially unconfined axial compression was measured by direct microscopic observation. Pressure, flow, and geometry data were input to a finite element model where hydraulic permeabilities in the disk axial and radial directions were determined. At less than 10% static compression, near free-swelling conditions, hydraulic permeability was nearly isotropic, with values corresponding to those of previous studies. With increasing static compression, hydraulic permeability decreased, but the radially directed permeability decreased more dramatically than the axially directed permeability such that strong anisotropy (a 10-fold difference between axial and radial directions) in the hydraulic permeability tensor was evident for static compression of 20-40%. Results correspond well with predictions of a previous microstructurally-based model for effects of tissue mechanical deformations on glycosaminoglycan architecture and cartilage hydraulic permeability. Findings inform understanding of structure-function relationships in cartilage matrix, and suggest several biomechanical roles for compression induced anisotropic hydraulic permeability in articular cartilage. PMID- 16271598 TI - Biomechanical assessment of tissue retrieved after in vivo cartilage defect repair: tensile modulus of repair tissue and integration with host cartilage. AB - Failure to restore the mechanical properties of tissue at the repair site and its interface with host cartilage is a common problem in tissue engineering procedures to repair cartilage defects. Quantitative in vitro studies have helped elucidate mechanisms underlying processes leading to functional biomechanical changes. However, biomechanical assessment of tissue retrieved from in vivo studies of cartilage defect repair has been limited to compressive tests. Analysis of integration following in vivo repair has relied on qualitative histological methods. The objectives of this study were to develop a quantitative biomechanical method to assess (1) the tensile modulus of repair tissue and (2) its integration in vivo, as well as determine whether supplementation of transplanted chondrocytes with IGF-I affected these mechanical properties. Osteochondral blocks were obtained from a previous 8 month study on the effects of IGF-I on chondrocyte transplantation in the equine model. Tapered test specimens were prepared from osteochondral blocks containing the repair/native tissue interface and adjacently located blocks of intact native tissue. Specimens were then tested in uniaxial tension. The tensile modulus of repair tissue averaged 0.65 MPa, compared to the average of 5.2 MPa measured in intact control samples. Integration strength averaged 1.2 MPa, nearly half the failure strength of intact cartilage samples, 2.7 MPa. IGF-I treatment had no detectable effects on these mechanical properties. This represents the first quantitative biomechanical investigation of the tensile properties of repair tissue and its integration strength in an in vivo joint defect environment. PMID- 16271599 TI - Age-sensitivity of time-related in vivo deformability of human lumbar motion segments and discs in pure centric tension. AB - The goal of this study was to document the effect of aging, sex and disc level on time-dependent in vivo tensile deformability of human lumbar-lumbosacral motion segments and discs in pure centric tension, when the contracting effect of muscles can be neglected. Elongations of segments L3-L4, L4-L5 and L5-S1 were measured during the usual suspension hydrotraction therapy of patients, by using a subaqual ultrasound measuring method reported in (Kurutz et al., 2002a, 2003). Patients were suspended cervically in lukewarm water for 20 min, loaded by 20-20 N lead weights on ankles. The mean initial elastic elongations (strains) of segments or discs were about 0.8 mm (10%) for patients under 40 years; 0.5 mm (6%) between 40-60 years; and 0.2 mm (3%) over 60 years. The mean final viscoelastic elongations were 1.5 mm (18%) under 40 years; 1.2 mm (15%) between 40-60 years; and 0.6 mm (7%) over 60 years. In the beginning/end of the treatment, patients of extended segments were on average 6/8 years younger than those with unextended ones. Based on the in vivo measured elongations, initial tensile stiffness was obtained in terms of aging, sex and disc level. For the above age-classes, the approximate mean tensile stiffness of less/more degenerated lumbar FSUs or discs were about 600/800, 800/1000 and 1800/2800 N/mm, respectively. A new terminology, the so-called age-sensitivity has been introduced as a value of 0.01-0.04 mm/year elongation capacity decrease per a year of aging, after the age of 35. No significant difference was found between sexes regarding age-dependence in tension. PMID- 16271600 TI - An approach to the simulation of fluid-structure interaction in the aortic valve. AB - A pair of finite element models has been employed to study the interaction of blood flow with the operation of the aortic valve. A three-dimensional model of the left ventricle with applied wall displacements has been used to generate data for the spatially and time-varying blood velocity profile across the aortic aperture. These data have been used as the inlet loading conditions in a three dimensional model of the aortic valve and its surrounding structures. Both models involve fluid-structure interaction and simulate the cardiac cycle as a dynamic event. Confidence in the models was obtained by comparison with data obtained in a pulse duplicator. The results show a circulatory flow being generated in the ventricle which produces a substantially axial flow through the aortic aperture. The aortic valve behaves in an essentially symmetric way under the action of this flow, so that the pressure difference across the leaflets is approximately uniform. This work supports the use of spatially uniform but temporally variable pressure distributions across the leaflets in dry or structural models of aortic valves. The study is a major advance through its use of truly three-dimensional geometry, spatially non-uniform loading conditions for the valve leaflets and the successful modelling of progressive contact of the leaflets in a fluid environment. PMID- 16271601 TI - Optimization model predictions for postural coordination modes. AB - This paper examines the ability of the dynamic optimization model to predict changes between in-phase and anti-phase postural modes of coordination and to evaluate influence of two particular environmental and intentional constraints on postural strategy. The task studied was based on an experimental paradigm that consisted in tracking a target motion with the head. An original optimal procedure was developed for cyclic problems to calculate hip and ankle angular trajectories during postural sway with a minimum torque change criterion. Optimization results give a good description of the sudden bifurcation phase between in-phase and anti-phase postural coordination modes in visual target tracking. Transition frequency and predicted effects of environmental and intentional constraints are also in line with experimental observations described in existing literature. In particular, these investigations pointed out that postural planning process can be related to the minimization of a dynamic cost criterion with an equilibrium constraint. In conclusion, the optimization technique is well suited for the prediction of postural modes of coordination and seems to offer many opportunities for better comprehension of neuromuscular movement control. PMID- 16271602 TI - A numerical method for the continuous spectrum biphasic poroviscoelastic model of articular cartilage. AB - A method for numerical solution of the continuous spectrum linear biphasic poroviscoelastic (BPVE) model of articular cartilage is presented. The method is based on an alternate formulation of the continuous spectrum stress-strain law that is implemented using Gaussian quadrature integration combined with quadratic interpolation of the strain history. For N time steps, the cost of the method is O(N). The method is applied to a finite difference solution of the one dimensional confined compression BPVE stress-relaxation problem. For a range of relaxation times that are representative of articular cartilage, accuracy of the method is demonstrated by direct comparison to a theoretical Laplace transform solution. PMID- 16271603 TI - Dynamic shoulder dynamometry: a way to develop delay onset muscle soreness in shoulder muscles. AB - A dynamic shoulder dynamometer has been designed to develop a vertical downward force over a measured range of motion, i.e. scapular elevation. The dynamometer is force-controlled to apply a required force within the range of motion. Calibration procedures were performed to assess the reliability of the force and displacement measurements. The system is designed to operate in isometric or eccentric conditions. For eccentric exercise the dynamometer has, for the first time, provided possibilities to elicit delay onset muscle soreness in shoulder muscles. In future, the apparatus will enable investigations of sensory-motor interactions in the shoulder region. PMID- 16271604 TI - Finite element analysis of moment-rotation relationships for human cervical spine. AB - A comprehensive, geometrically accurate, nonlinear C0-C7 FE model of head and cervical spine based on the actual geometry of a human cadaver specimen was developed. The motions of each cervical vertebral level under pure moment loading of 1.0 Nm applied incrementally on the skull to simulate the movements of the head and cervical spine under flexion, tension, axial rotation and lateral bending with the inferior surface of the C7 vertebral body fully constrained were analysed. The predicted range of motion (ROM) for each motion segment were computed and compared with published experimental data. The model predicted the nonlinear moment-rotation relationship of human cervical spine. Under the same loading magnitude, the model predicted the largest rotation in extension, followed by flexion and axial rotation, and least ROM in lateral bending. The upper cervical spines are more flexible than the lower cervical levels. The motions of the two uppermost motion segments account for half (or even higher) of the whole cervical spine motion under rotational loadings. The differences in the ROMs among the lower cervical spines (C3-C7) were relatively small. The FE predicted segmental motions effectively reflect the behavior of human cervical spine and were in agreement with the experimental data. The C0-C7 FE model offers potentials for biomedical and injury studies. PMID- 16271620 TI - The natural toxin juglone causes degradation of p53 and induces rapid H2AX phosphorylation and cell death in human fibroblasts. AB - Juglone (5-hydroxy-1,4-naphtoquinone) is a natural toxin produced by walnut trees. In this study we show that juglone differentially reduces viability of human cells in culture. Normal fibroblast were found to be especially sensitive to juglone and lost viability primarily through a rapid apoptotic and necrotic response. This response may have been triggered by DNA damage since juglone induced a rapid and strong phosphorylation of H2AX in all phases of the cell cycle. Furthermore, juglone inhibits mRNA synthesis in human fibroblasts in a dose-dependent manner. Surprisingly, juglone caused a drastic reduction of the basal level of p53 in human fibroblasts and this loss could not be fully rescued by proteasome and calpain I inhibitors. However, when cells were pretreated with UV light or ionizing radiation, juglone was not able to reduce the cellular levels of activated p53. Our results show that juglone has multiple effects on cells such as the induction of DNA damage, inhibition of transcription, reduction of p53 protein levels and the induction of cell death. PMID- 16271621 TI - Arsenite enhances tumor necrosis factor-alpha-induced expression of vascular cell adhesion molecule-1. AB - Epidemiological studies demonstrated a high association of vascular diseases with arsenite exposure. We hypothesize that arsenite potentiates the effect of proinflammatory cytokines on vascular endothelial cells, and hence contributes to atherosclerosis. In this study, we investigated the effect of arsenite and its induction of glutathione (GSH) on vascular cell adhesion molecule-1 (VCAM-1) protein expression in human umbilical vein endothelial cells (HUVECs) in response to tumor necrosis factor-alpha (TNF-alpha), a typical proinflammatory cytokine. Our study demonstrated that arsenite pretreatment potentiated the TNF-alpha induced VCAM-1 expression with up-regulations of both activator protein-1 (AP-1) and nuclear factor-kappaB (NF-kappaB). To elucidate the role of GSH in regulation of AP-1, NF-kappaB, and VCAM-1 expression, we employed l-buthionine (S,R) sulfoximine (BSO), a specific gamma-glutamylcysteine synthetase (gamma-GCS) inhibitor, to block intracellular GSH synthesis. Our investigation revealed that, by depleting GSH, arsenite attenuated the TNF-alpha-induced VCAM-1 expression as well as a potentiation of AP-1 and an attenuation of NF-kappaB activations by TNF alpha. Moreover, we found that depletion of GSH would also attenuate the TNF alpha-induced VCAM-1 expression with a down-regulation of the TNF-alpha-induced NF-kappaB activation and without significant effect on AP-1. On the other hand, the TNF-alpha-induced VCAM-1 expression could be completely abolished by inhibition of AP-1 or NF-kappaB activity, suggesting that activation of both AP-1 and NF-kappaB was necessary for VCAM-1 expression. In summary, we demonstrate that arsenite enhances the TNF-alpha-induced VCAM-1 expression in HUVECs via regulation of AP-1 and NF-kappaB activities in a GSH-sensitive manner. Our present study suggested a potential mechanism for arsenite in the induction of vascular inflammation and vascular diseases via modulating the actions of proinflammatory cytokines. PMID- 16271622 TI - Macrophages are involved in hexachlorobenzene-induced adverse immune effects. AB - Hexachlorobenzene (HCB) is a persistent environmental pollutant that causes adverse immune effects in man and rat. The Brown Norway (BN) rat is very susceptible to HCB-induced immunopathology and oral exposure causes inflammatory skin and lung lesions, splenomegaly, lymph node (LN) enlargement, and increased serum levels of IgE and anti-ssDNA IgM. T cells play an important role but do not account for all adverse effects induced by HCB. Macrophages are probably also important and the relationship between macrophages and T cells was further investigated. To eliminate macrophages clodronate-liposomes were used. Furthermore, a kinetic study was performed to obtain insight in the early phase of the HCB-induced immune response. Also, experiments were performed to detect specific memory T cells. Therefore, an adoptive transfer study was performed. Our results indicate that macrophages are indeed involved in HCB-induced skin lesions, lung eosinophilia, and elevation of IgM against ssDNA. Kinetics showed that both skin and lung lesions appeared early after exposure. Moreover, immune effects could not be adaptively transferred. Thus, both macrophages and T cells are involved in HCB-induced immune effects but HCB exposure does not lead to specific T cell sensitization. Presumably, HCB exposure induces macrophage activation, thereby generating adjuvant signals that polyclonally stimulate T cells. Together, these events may lead to the observed immunopathology in BN rats. PMID- 16271623 TI - Anticonvulsant treatment of sarin-induced seizures with nasal midazolam: an electrographic, behavioral, and histological study in freely moving rats. AB - Centrally mediated seizures and convulsions are common consequences of exposure to organophosphates (OPs). These seizures rapidly progress to status epilepticus (SE) and contribute to profound brain injury. Effective management of these seizures is critical for minimization of brain damage. Nasal application of midazolam (1.5 mg/kg) after 5 min of sarin-induced electrographic seizure activity (EGSA) ameliorated EGSA and convulsive behavior (238 +/- 90 s). Identical treatment after 30 min was not sufficient to ameliorate ECoG paradoxical activity and convulsive behavior. Nasal midazolam (1.5 mg/kg), together with scopolamine (1 mg/kg, im) after 5 min of EGSA, exerted a powerful and rapid anticonvulsant effect (53 +/- 10 s). Delaying the same treatment to 30 min of EGSA leads to attenuation of paroxysmal ECoG activity in all cases but total cessation of paroxysmal activity was not observed in most animals tested. Cognitive tests utilizing the Morris Water Maze demonstrated that nasal midazolam alone or together with scopolamine (im), administered after 5 min of convulsions, abolished the effect of sarin on learning. Both these treatments, when given after 30 min of convulsions, only decreased the sarin-induced learning impairments. Whereas rats which were not subject to the anticonvulsant agents did not show any memory for the platform location, both treatments (at 5 min as well as at 30 min) completely abolished the memory deficits. Both treatments equally blocked the impairment of reversal learning when given at 5 min. However, when administered after 30 min, midazolam alone reversed the impairments in reversal learning, while midazolam with scopolamine did not. Rats exposed to sarin and treated with the therapeutic regimen with the exclusion of midazolam exhibited severe brain lesions that encountered the hippocampus, pyriform cortex, and thalamus. Nasal midazolam at 5 min prevented brain damage, while delaying the midazolam treatment to 30 min of EGSA resulted in brain damage. The addition of scopolamine to midazolam did not alter the above observation. In summary, nasal midazolam treatment briefly after initiation of OP-induced seizure leads to cessation of EGSA and prevented brain lesions and behavioral deficiencies in the rat model. PMID- 16271625 TI - Crunch time for summit on avian and human influenza. PMID- 16271624 TI - Time-dependent effect of p-aminophenol (PAP) toxicity in renal slices and development of oxidative stress. AB - p-Aminophenol (PAP), a metabolite of acetaminophen, is nephrotoxic. This study investigated PAP-mediated changes as a function of time that occur prior to loss of membrane integrity. Experiments further evaluated the development of oxidative stress by PAP. Renal slices from male Fischer 344 (F344) rats (N = 4-6) were exposed to 0.1, 0.25, and 0.5 mM PAP for 15-120 min under oxygen and constant shaking at 37 degrees C. Pyruvate-stimulated gluconeogenesis, adenine nucleotide levels, and total glutathione (GSH) levels were diminished in a concentration- and time-dependent manner prior to detection of a rise in lactate dehydrogenase (LDH) leakage. Glutathione disulfide (GSSG) levels were increased by PAP suggesting the induction of oxidative stress. Western blot analysis confirmed a rise in 4-hydroxynonenal (4-HNE)-adducted proteins in tissues exposed to 0.1 and 0.25 mM PAP for 90 min. The appearance of 4-HNE-adducted proteins at the 0.1 mM concentration of PAP occurred prior to development of increased LDH leakage. Pretreatment with 1 mM glutathione (GSH) for 30 min only partially reduced PAP toxicity as LDH values were less severely depleted relative to tissues not pretreated with GSH. In contrast, pretreatment for 15 min with 2 mM ascorbic acid completely protected against PAP toxicity. Further studies showed that ascorbic acid pretreatment prevented PAP-mediated depletion of GSH. In summary, PAP rapidly depletes GSH and adenine nucleotides and inhibits gluconeogenesis prior to a rise in LDH leakage. PAP induces oxidative stress as indicated by an increase in GSSG and 4-HNE-adducted proteins. Ascorbic acid pretreatment prevents PAP toxicity by maintaining GSH status. PMID- 16271626 TI - Gates leads on malaria funding. PMID- 16271627 TI - The long and winding road towards a tobacco-free world. PMID- 16271628 TI - Something old, something new: beta blockers and clopidogrel in acute myocardial infarction. PMID- 16271629 TI - A farewell to body-mass index? PMID- 16271630 TI - Congenital varicella syndrome in the third trimester. PMID- 16271631 TI - Is public health coercive health? PMID- 16271633 TI - Hitoshi Oshitani: watching out for an influenza pandemic. PMID- 16271634 TI - Typhoid and paratyphoid fever. PMID- 16271635 TI - Typhoid and paratyphoid fever. PMID- 16271636 TI - Typhoid and paratyphoid fever. PMID- 16271637 TI - Judging psychiatric disorders in refugees. PMID- 16271638 TI - Breast cancer. PMID- 16271640 TI - Improving health workers' performance in low-resource settings. PMID- 16271642 TI - Addition of clopidogrel to aspirin in 45,852 patients with acute myocardial infarction: randomised placebo-controlled trial. AB - BACKGROUND: Despite improvements in the emergency treatment of myocardial infarction (MI), early mortality and morbidity remain high. The antiplatelet agent clopidogrel adds to the benefit of aspirin in acute coronary syndromes without ST-segment elevation, but its effects in patients with ST-elevation MI were unclear. METHODS: 45,852 patients admitted to 1250 hospitals within 24 h of suspected acute MI onset were randomly allocated clopidogrel 75 mg daily (n=22,961) or matching placebo (n=22,891) in addition to aspirin 162 mg daily. 93% had ST-segment elevation or bundle branch block, and 7% had ST-segment depression. Treatment was to continue until discharge or up to 4 weeks in hospital (mean 15 days in survivors) and 93% of patients completed it. The two prespecified co-primary outcomes were: (1) the composite of death, reinfarction, or stroke; and (2) death from any cause during the scheduled treatment period. Comparisons were by intention to treat, and used the log-rank method. This trial is registered with ClinicalTrials.gov, number NCT00222573. FINDINGS: Allocation to clopidogrel produced a highly significant 9% (95% CI 3-14) proportional reduction in death, reinfarction, or stroke (2121 [9.2%] clopidogrel vs 2310 [10.1%] placebo; p=0.002), corresponding to nine (SE 3) fewer events per 1000 patients treated for about 2 weeks. There was also a significant 7% (1-13) proportional reduction in any death (1726 [7.5%] vs 1845 [8.1%]; p=0.03). These effects on death, reinfarction, and stroke seemed consistent across a wide range of patients and independent of other treatments being used. Considering all fatal, transfused, or cerebral bleeds together, no significant excess risk was noted with clopidogrel, either overall (134 [0.58%] vs 125 [0.55%]; p=0.59), or in patients aged older than 70 years or in those given fibrinolytic therapy. INTERPRETATION: In a wide range of patients with acute MI, adding clopidogrel 75 mg daily to aspirin and other standard treatments (such as fibrinolytic therapy) safely reduces mortality and major vascular events in hospital, and should be considered routinely. PMID- 16271643 TI - Early intravenous then oral metoprolol in 45,852 patients with acute myocardial infarction: randomised placebo-controlled trial. AB - BACKGROUND: Despite previous randomised trials of early beta-blocker therapy in the emergency treatment of myocardial infarction (MI), uncertainty has persisted about the value of adding it to current standard interventions (eg, aspirin and fibrinolytic therapy), and the balance of potential benefits and hazards is still unclear in high-risk patients. METHODS: 45,852 patients admitted to 1250 hospitals within 24 h of suspected acute MI onset were randomly allocated metoprolol (up to 15 mg intravenous then 200 mg oral daily; n=22,929) or matching placebo (n=22,923). 93% had ST-segment elevation or bundle branch block, and 7% had ST-segment depression. Treatment was to continue until discharge or up to 4 weeks in hospital (mean 15 days in survivors) and 89% completed it. The two prespecified co-primary outcomes were: (1) composite of death, reinfarction, or cardiac arrest; and (2) death from any cause during the scheduled treatment period. Comparisons were by intention to treat, and used the log-rank method. This study is registered with ClinicalTrials.gov, number NCT 00222573. FINDINGS: Neither of the co-primary outcomes was significantly reduced by allocation to metoprolol. For death, reinfarction, or cardiac arrest, 2166 (9.4%) patients allocated metoprolol had at least one such event compared with 2261 (9.9%) allocated placebo (odds ratio [OR] 0.96, 95% CI 0.90-1.01; p=0.1). For death alone, there were 1774 (7.7%) deaths in the metoprolol group versus 1797 (7.8%) in the placebo group (OR 0.99, 0.92-1.05; p=0.69). Allocation to metoprolol was associated with five fewer people having reinfarction (464 [2.0%] metoprolol vs 568 [2.5%] placebo; OR 0.82, 0.72-0.92; p=0.001) and five fewer having ventricular fibrillation (581 [2.5%] vs 698 [3.0%]; OR 0.83, 0.75-0.93; p=0.001) per 1000 treated. Overall, these reductions were counterbalanced by 11 more per 1000 developing cardiogenic shock (1141 [5.0%] vs 885 [3.9%]; OR 1.30, 1.19-1.41; p<0.00001). This excess of cardiogenic shock was mainly during days 0-1 after admission, whereas the reductions in reinfarction and ventricular fibrillation emerged more gradually. Consequently, the overall effect on death, reinfarction, arrest, or shock was significantly adverse during days 0-1 and significantly beneficial thereafter. There was substantial net hazard in haemodynamically unstable patients, and moderate net benefit in those who were relatively stable (particularly after days 0-1). INTERPRETATION: The use of early beta-blocker therapy in acute MI reduces the risks of reinfarction and ventricular fibrillation, but increases the risk of cardiogenic shock, especially during the first day or so after admission. Consequently, it might generally be prudent to consider starting beta-blocker therapy in hospital only when the haemodynamic condition after MI has stabilised. PMID- 16271644 TI - Diabetes and associated disorders in Cambodia: two epidemiological surveys. AB - BACKGROUND: The Asia-Pacific region is thought to be severely affected by diabetes. However, reliable, standardised data on prevalence and characteristics of glucose intolerance in Asian populations remain sparse. We describe the results of two field surveys undertaken in Cambodia in 2004. METHODS: 2246 randomly selected adults aged 25 years and older were examined in two communities, one rural (Siemreap) and one semi-urban (Kampong Cham). The diagnosis of diabetes and impaired glucose tolerance was based on 2-h blood glucose estimation using criteria recommended by the latest report of a WHO Expert Group. Blood pressure, anthropometry, habitual diet, and other relevant characteristics were also recorded. FINDINGS: Prevalence of diabetes was 5% in Siemreap and 11% in Kampong Cham. Prevalence of impaired glucose tolerance was 10% in Siemreap and 15% in Kampong Cham. About two-thirds of all cases of diabetes were undiagnosed before the survey. Prevalence of hypertension was 12% at Siemreap and 25% at Kampong Cham. People in Kampong Cham had higher estimates of central obesity than those in Siemreap. INTERPRETATION: Diabetes and hypertension are not uncommon in Cambodia. A quarter of all adults in the chosen suburban community had some degree of glucose intolerance. Since Cambodian society is relatively poor, and lifestyle is fairly traditional by international standards, these findings are unexpected. PMID- 16271645 TI - Obesity and the risk of myocardial infarction in 27,000 participants from 52 countries: a case-control study. AB - BACKGROUND: Obesity is a major risk factor for cardiovascular disease, but the most predictive measure for different ethnic populations is not clear. We aimed to assess whether markers of obesity, especially waist-to-hip ratio, would be stronger indicators of myocardial infarction than body-mass index (BMI), the conventional measure. METHODS: We did a standardised case-control study of acute myocardial infarction with 27 098 participants in 52 countries (12,461 cases and 14,637 controls) representing several major ethnic groups. We assessed the relation between BMI, waist and hip circumferences, and waist-to-hip ratio to myocardial infarction overall and for each group. FINDINGS: BMI showed a modest and graded association with myocardial infarction (OR 1.44, 95% CI 1.32-1.57 top quintile vs bottom quintile before adjustment), which was substantially reduced after adjustment for waist-to-hip ratio (1.12, 1.03-1.22), and non-significant after adjustment for other risk factors (0.98, 0.88-1.09). For waist-to-hip ratio, the odds ratios for every successive quintile were significantly greater than that of the previous one (2nd quintile: 1.15, 1.05-1.26; 3rd quintile: 1.39; 1.28-1.52; 4th quintile: 1.90, 1.74-2.07; and 5th quintiles: 2.52, 2.31-2.74 [adjusted for age, sex, region, and smoking]). Waist (adjusted OR 1.77; 1.59 1.97) and hip (0.73; 0.66-0.80) circumferences were both highly significant after adjustment for BMI (p<0.0001 top vs bottom quintiles). Waist-to-hip ratio and waist and hip circumferences were closely (p<0.0001) associated with risk of myocardial infarction even after adjustment for other risk factors (ORs for top quintile vs lowest quintiles were 1.75, 1.33, and 0.76, respectively). The population-attributable risks of myocardial infarction for increased waist-to-hip ratio in the top two quintiles was 24.3% (95% CI 22.5-26.2) compared with only 7.7% (6.0-10.0) for the top two quintiles of BMI. INTERPRETATION: Waist-to-hip ratio shows a graded and highly significant association with myocardial infarction risk worldwide. Redefinition of obesity based on waist-to-hip ratio instead of BMI increases the estimate of myocardial infarction attributable to obesity in most ethnic groups. PMID- 16271646 TI - Role of the mitochondrial DNA 16184-16193 poly-C tract in type 2 diabetes. AB - Recent evidence suggests that polymorphic genetic variation in the non-coding region of mitochondrial DNA (the 16184-16193 polycytosine [poly-C] tract) contributes to the cause of type 2 diabetes, but previous studies only just reached significance. We aimed to investigate this association. We compared patients with type 2 diabetes (n=992) with two independent control groups (n=536, n=1029) from the UK, and saw no difference in the frequency of the 16184-16193 poly-C tract. This finding was confirmed by a meta-analysis of European studies of 1455 patients and 3132 controls (odds ratio 1.16, 95% CI 0.94-1.44). Genetic variation of the 16184-16193 poly-C tract is unlikely to have a major role in the cause of type 2 diabetes. PMID- 16271647 TI - Broken needles. PMID- 16271648 TI - Guillain-Barre syndrome. AB - Guillain-Barre syndrome consists of at least four subtypes of acute peripheral neuropathy. Major advances have been made in understanding the mechanisms of some of the subtypes. The histological appearance of the acute inflammatory demyelinating polyradiculoneuropathy (AIDP) subtype resembles experimental autoimmune neuritis, which is predominantly caused by T cells directed against peptides from the myelin proteins P0, P2, and PMP22. The role of T-cell-mediated immunity in AIDP remains unclear and there is evidence for the involvement of antibodies and complement. Strong evidence now exists that axonal subtypes of Guillain-Barre syndrome, acute motor axonal neuropathy (AMAN), and acute motor and sensory axonal neuropathy (AMSAN), are caused by antibodies to gangliosides on the axolemma that target macrophages to invade the axon at the node of Ranvier. About a quarter of patients with Guillain-Barre syndrome have had a recent Campylobacter jejuni infection, and axonal forms of the disease are especially common in these people. The lipo-oligosaccharide from the C jejuni bacterial wall contains ganglioside-like structures and its injection into rabbits induces a neuropathy that resembles acute motor axonal neuropathy. Antibodies to GM1, GM1b, GD1a, and GalNac-GD1a are in particular implicated in acute motor axonal neuropathy and, with the exception of GalNacGD1a, in acute motor and sensory axonal neuropathy. The Fisher's syndrome subtype is especially associated with antibodies to GQ1b, and similar cross-reactivity with ganglioside structures in the wall of C jejuni has been discovered. Anti-GQ1b antibodies have been shown to damage the motor nerve terminal in vitro by a complement-mediated mechanism. Results of international randomised trials have shown equivalent efficacy of both plasma exchange and intravenous immunoglobulin, but not corticosteroids, in hastening recovery from Guillain-Barre syndrome. Further research is needed to discover treatments to prevent 20% of patients from being left with persistent and significant disability. PMID- 16271649 TI - Preventing chronic diseases: taking stepwise action. AB - The scientific knowledge to achieve a new global goal for the prevention of chronic diseases--a 2% yearly reduction in rates of death from chronic disease over and above projected declines during the next 10 years--already exists. However, many low-income and middle-income countries must deal with the practical realities of limited resources and a double burden of infectious and chronic diseases. This paper presents a novel planning framework that can be used in these contexts: the stepwise framework for preventing chronic diseases. The framework offers a flexible and practical public health approach to assist ministries of health in balancing diverse needs and priorities while implementing evidence-based interventions such as those recommended by the WHO Framework Convention on Tobacco Control and the WHO Global Strategy on Diet, Physical Activity and Health. Countries such as Indonesia, the Philippines, Tonga, and Vietnam have applied the stepwise planning framework: their experiences illustrate how the stepwise approach has general applicability to solving chronic disease problems without sacrificing specificity for any particular country. PMID- 16271650 TI - Pseudo-pseudo Meigs' syndrome. PMID- 16271651 TI - Depression following myocardial infarction. PMID- 16271652 TI - Physicians' satisfaction with a collaborative disease management program for late life depression in primary care. AB - OBJECTIVE: This study describes physicians' satisfaction with care for patients with depression before and after the implementation of a primary care-based collaborative care program. METHOD: Project Improving Mood, Promoting Access to Collaborative Treatment for late-life depression (IMPACT) is a multisite, randomized controlled trial comparing a primary care-based collaborative disease management program for late-life depression with care as usual. A total of 450 primary care physicians at 18 participating clinics participated in a satisfaction survey before and 12 months after IMPACT initiation. The preintervention survey focused on physicians' satisfaction with current mental health resources and ability to provide depression care. The postintervention survey repeated these and added questions about physician's experience with the IMPACT collaborative care model. RESULTS: Before intervention, about half (54%) of the participating physicians were satisfied with resources to treat patients with depression. After intervention, more than 90% reported the intervention as helpful in treating patients with depression and 82% felt that the intervention improved patients' clinical outcomes. Participating physicians identified proactive patient follow-up and patient education as the most helpful components of the IMPACT model. CONCLUSIONS: Physicians perceived a substantial need for improving depression treatment in primary care. They were very satisfied with the IMPACT collaborative care model for treating depressed older adults and felt that similar care management models would also be helpful for treating other chronic medical illnesses. PMID- 16271653 TI - Posttraumatic stress disorder in primary care: prevalence and relationships with physical symptoms and medical utilization. AB - OBJECTIVE: This study estimates the prevalence of posttraumatic stress disorder (PTSD) and describes the relationships among PTSD status and health indices in a civilian primary care patient sample. METHODS: Participants (N = 232) completed a paper-and-pencil survey of life events, PTSD symptoms, physical symptoms and health functioning. Utilization was assessed from medical records. RESULTS: Nine percent of the participants met the criteria for full PTSD (based on Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition criteria) and another 25% were defined as partial PTSD. The full-PTSD group evidenced higher rates of medical utilization, more intense physical symptoms and poorer health functioning than the no-PTSD group. The partial-PTSD group more closely resembled the full PTSD group. CONCLUSIONS: This study, although limited by sample size and diagnosis by questionnaire vs. diagnostic interview, suggests research directions for enhancing our understanding of PTSD among civilian primary care patients and for developing appropriate interventions that can be conducted in the primary care setting. PMID- 16271654 TI - Feasibility and acceptability of a telephone psychotherapy program for depressed adults treated in primary care. AB - OBJECTIVE: Telephone psychotherapy is an emerging model of care that appears feasible for extending the reach of evidence-based psychotherapy treatment without accruing the full costs of traditional office-based, mental health care. This manuscript describes the development, implementation and acceptance of a 12 month telephone psychotherapy program (TPP) for depressed adults not fully responding to standard antidepressant treatment in primary care. METHOD: The TPP combined a population-based medication monitoring and information system with a structured cognitive-behavioral treatment (CBT) program. The TPP included 8-12 telephone sessions (eight core CBT sessions and three to four clinical booster sessions) delivered by a master-level therapist working in tandem with each patient's primary care physician (PCP). RESULTS: The TPP was well accepted (i.e., 80% completed the core program) by a population-based sample of adult primary care patients initiating antidepressant treatment. The mean duration of core telephone psychotherapy sessions was approximately 31 min during acute-phase treatment (0-6 months). Eighty-two percent of TPP patients maintained contact with their therapist during maintenance-phase treatment (6-12 months). CONCLUSIONS: The practical and efficient nature of this TPP appears to sidestep many of the treatment barriers encountered in traditional office-based care. Implementation of this TPP program in other primary care settings may be valuable for enhancing standard pharmacotherapy treatment of adult depression, especially among populations facing greater barriers of care. PMID- 16271655 TI - Depression following myocardial infarction: first-ever versus ongoing and recurrent episodes. AB - BACKGROUND: Depression following myocardial infarction (MI) can be a first-ever episode for some, whereas for others, it may represent a recurrent episode or one that was present at the onset of the infarction. We investigated if there are differences in pre- and post-MI characteristics between these subtypes. METHODS: Four hundred sixty-eight patients admitted for an MI were assessed for the presence of an ICD-10 depressive disorder following MI. A comparison was made between first-ever and ongoing or recurrent depression on demographic and cardiac data, personality, and depression characteristics. RESULTS: Depressive disorder during the first post-MI year was present in 25.4% of the MI patients (n = 119), and almost half were ongoing or recurrent (n = 53, 44.5%). Recurrent and ongoing depression was related to high neuroticism (Z = 2.77, P < .01), whereas first ever depression was associated with MI severity (poor left ventricular ejection fraction: Z = 1.64, P = .05; PTCA or CABG during hospitalization: Z = 1.88, P = .03; arrhythmic events: Z = 1.49, P = .06). CONCLUSIONS: Our results suggest that in the first-ever post-MI depression cases, depression may be triggered by the severity of the MI, whereas ongoing and recurrent depression is more related to personality. Future research should address the question whether these subtypes of depression differ in cardiovascular prognosis and response to psychiatric treatment. PMID- 16271656 TI - Unmet need for medical care among homeless adults with serious mental illness. AB - OBJECTIVE: The objective of this study was to determine the rates and predictors of unmet medical needs among homeless adults with serious mental illness entering a community-based case management program and those of receipt of medical care in the subsequent 3-month period. METHODS: We analyzed baseline and follow-up data for 7213 homeless clients in the multisite Access to Community Care and Effective Services and Supports program. RESULTS: Overall, 43.6% of the sample reported having an unmet need for medical care at baseline; of these subjects, only 36.1% received medical services during the 3 months following program entry. Using multivariate logistic regression modeling, we found that, at baseline, independent correlates of an unmet medical need included lower educational level, increased depressive and psychotic symptoms and greater number of potentially competing needs. None of these variables, however, adversely affected the likelihood of receiving medical care during follow-up. Factors associated with receiving medical services in the 3 months following program entry included receiving outpatient psychiatric services and reporting stronger therapeutic alliance with one's case manager. CONCLUSIONS: Collaborative case management may play an important role in meeting mentally ill homeless persons' medical needs. Greater efforts are needed to identify and link at-risk clients with appropriate medical services. PMID- 16271657 TI - Obsessive-compulsive disorder in a dermatology outpatient clinic. AB - OBJECTIVE: The aims of present study were to (a) to determine the prevalence of obsessive-compulsive disorder (OCD) in dermatological patients, (b) to determine the possible relationship between dermatological lesions and OCD and (c) to determine the clinical and phenomenological features of the OCD subgroup. METHOD: The sample consisted of 166 out of 250 consecutively presenting dermatological patients who agreed to participate in the study. The subjects were assessed with the Structured Clinical Interview for DSM-IV Turkish Version (SCID-I) and also completed the Yale-Brown Obsessive Compulsive Scale (Y-BOCS). RESULTS: Of the whole sample, 41 (24.7%) met DSM-IV criteria for OCD. Only 14.6% of them had previously been diagnosed as OCD. The mean score of Y-BOCS in the OCD group was 17.05 +/- 9.75. The most common obsessions were contamination (61%) and pathologic doubt (53.7%), while washing (61%) and checking (51.2%) were the most frequent compulsions. Those suffering from diseases of sebaceous glands were the only group that showed a significant difference between the OCD and non-OCD group. CONCLUSION: There is a high prevalence of OCD in dermatological patients, although the nature of the relationship between OCD and dermatology has not previously been ascertained. Genetic-based studies and future researches focused on individual anxiety, and sensitivity may provide information that better explains this relationship. PMID- 16271658 TI - Depression and anxiety in patients with hepatitis C: prevalence, detection rates and risk factors. AB - OBJECTIVE: We examined a group of patients awaiting interferon treatment for hepatitis C to estimate the prevalence and detection rates of and risk factors for mood disorders. METHODS: The Structured Clinical Interview for DSM-IV Axis I Disorders: Clinician Version was used to detect psychiatric disorder. Self completion instruments were used to rate symptom severity, subjective cognitive function, work and social adjustment, stigma, acceptance of illness and treatment satisfaction. RESULTS: The 90 participants included 23 women (26%); 33 (37%) had contracted hepatitis C iatrogenically, 42 (47%) through injecting drug use and the remainder (17%) were of unknown origin. There was a 28% 1-month prevalence of depressive disorders, 72% of whom were previously undiagnosed, and a 24% prevalence of anxiety disorders, 86% previously undiagnosed. Current methadone maintenance was strongly associated with risk of depression (odds ratio, 5.0; 95% CI, 1.08-23.0). After adjustment for age and sex, depression was associated with poorer work and social adjustment, lower acceptance of illness, higher illness stigma, poorer reported thinking and concentration, and higher levels of subjective physical symptoms (all P < .05). Anxiety disorders were uncorrelated with any risk factor. CONCLUSIONS: Depression and anxiety have high prevalences in hepatitis C, and are largely undetected and treated. Depression, but not anxiety, is associated with adverse experiences of illness. PMID- 16271659 TI - Clinical correlates of CES-D depressive symptom ratings in an MS population. AB - OBJECTIVE: In multiple sclerosis (MS), depression rating scales may be used as case-finding instruments for depressive disorders, but depressive symptom ratings may have clinical implications beyond their case-finding role. The objective of this analysis was to explore this possibility by carrying out descriptive analyses of symptom ratings obtained using the Center for Epidemiological Studies Depression Rating Scale (CES-D) in an MS clinic population. METHOD: The analysis used cross-sectional baseline data collected from 589 subjects enrolled in a prospective cohort study. Data collection included demographic and clinical information including Extended Disability Status Scale ratings, a 54-item MS Quality of Life Scale and the Fatigue Impact Scale. RESULTS: Across a spectrum of CES-D scores, correlations with other health indicators were observed. Depressive symptoms were higher in more disabled subjects. CES-D scores were correlated with the emotional well-being dimension of quality of life and with a social fatigue impact dimension. Alternative scoring of the CES-D had a negligible impact on the pattern of correlation. CONCLUSIONS: CES-D ratings appear to be interpretable beyond the traditional yes/no categorization used in case finding. CES-D ratings are predictive of other clinical parameters in domains relevant to mental health. PMID- 16271660 TI - Higher stakes ahead for cultural competence. AB - Cultural competence, as a tool to overcome disparities in mental healthcare to ethnic minorities, is facing unprecedented challenges. Providers are turning to cultural competence as one of the few resources available for badly needed information and strategies for improving clinical care. This essay provides background information about this issue and addresses two broad questions. First, are cultural competence programs ready to meet the test? And second, what are the likely consequences if these programs don't adequately meet expectations? This article also reviews what is at stake for clients, health care providers, and advocates of cultural competency in using this paradigm to address the disparities crisis. Recommendations for future progress in this field are discussed. PMID- 16271661 TI - Mood swing from severe depression to mania following acute alteration of thyroid status. PMID- 16271662 TI - Case report: repetitive autocastration secondary to severe personality disorder. PMID- 16271664 TI - Do emergency department blood cultures change practice in patients with pneumonia? AB - STUDY OBJECTIVE: Although it is considered standard of care to obtain blood cultures on patients hospitalized for pneumonia, several studies have questioned the utility and cost-effectiveness of this practice. The objective of this study is to determine the impact of emergency department (ED) blood cultures on antimicrobial therapy for patients with pneumonia. METHODS: We performed a prospective, observational, cohort study of consecutive adult (age > or =18 years) patients treated at an urban university ED between February 1, 2000 and February 1, 2001. Inclusion criteria were radiographic evidence of pneumonia, clinical evidence of pneumonia, and blood culture obtained. Blood cultures were classified as positive, negative, or contaminant based on previously established criteria. Additionally, data were collected on antimicrobial sensitivities, empiric antibiotic therapy, antibiotic changes, and reasons for changes. RESULTS: There were 3,926 ED visits with blood cultures obtained for any reason, of which 3,762 (96%) were available for review. Of these, 414 of 3,762 (11%) patients met pneumonia study inclusion criteria, and blood cultures identified 29 of 414 (7.0%) patients with true bacteremia. In the 414 patients, blood culture results altered therapy for 15 patients (3.6%) with suspected pneumonia, of which 11 (2.7%) patients had their coverage narrowed; only 4 (1.0%) patients had their coverage broadened because of resistance to empiric therapy. For the 11 patients with bacteremia whose therapy was not altered, culture results actually supported narrowing therapy in 8 (1.9%) cases, but this was not done. CONCLUSION: Blood cultures rarely altered therapy for patients presenting to the ED with pneumonia. More discriminatory blood culture use may potentially reduce resource utilization. PMID- 16271665 TI - Fatal community-associated methicillin-resistant Staphylococcus aureus pneumonia in an immunocompetent young adult. AB - Severe pneumonia caused by community-associated methicillin-resistant Staphylococcus aureus (MRSA) was reported in children soon after this pathogen emerged in the United States in the 1990s. Genes for Panton Valentine leukocidin, which are present in the majority of community-associated MRSA, are thought to enhance the ability of S aureus to cause necrotizing pneumonia. Despite the rapid spread throughout the United States of community-associated MRSA and related skin and soft-tissue infections, reports of severe pneumonia in adults have been rare. We describe a case of a healthy young adult who initially was treated as an outpatient with levofloxacin for what appeared to be typical community-acquired pneumonia. He soon returned to the emergency department (ED) with rapidly fatal necrotizing pneumonia, associated with hemoptysis, leukopenia, and sepsis syndrome, that was caused by community-associated MRSA carrying genes for Panton Valentine leukocidin. This case highlights the typical features of this form of pneumonia and the need to consider MRSA when evaluating and treating severe pneumonia in the ED. It also raises the question of whether the incidence of this form of pneumonia might be increasing in communities with a high prevalence of community-associated MRSA and whether current pneumonia treatment guidelines should be modified. PMID- 16271663 TI - Abnormal physiological conditions due to hypersympathetic activity in psychiatric emergency patients. PMID- 16271666 TI - Images in emergency medicine. Laryngeal papilloma. PMID- 16271667 TI - Images in emergency medicine. Parinaud's oculoglandular syndrome. PMID- 16271668 TI - Blood cultures for community-acquired pneumonia: can we hit the target without a shotgun? PMID- 16271669 TI - The Joint Commission on Accreditation of Healthcare Organizations and Center for Medicare and Medicaid Services community-acquired pneumonia initiative: what went wrong? PMID- 16271670 TI - Performance characteristics of clinical diagnosis, a clinical decision rule, and a rapid influenza test in the detection of influenza infection in a community sample of adults. AB - STUDY OBJECTIVE: The accurate diagnosis of influenza remains a diagnostic dilemma. We examine the performance of various strategies for diagnosing influenza infection in an unselected sample of adults during influenza season. METHODS: Consecutive adults presenting to a university emergency department or urgent care clinic between January and March 2002 with acute respiratory complaints were eligible for this prospective observational study. The performance of clinician judgment, a rapid influenza test, and a clinical prediction rule in predicting influenza infection was evaluated using referent standard of reverse transcriptase polymerase chain reaction. Statistical significance was assessed using McNemar's test of proportions. RESULTS: Fifty three of 258 (21%) patients had a positive influenza reverse transcriptase polymerase chain reaction test. Overall, clinician judgment showed sensitivity of 29% (95% confidence interval [CI] 18% to 43%) and specificity of 92% (95% CI 87% to 95%). The rapid influenza test showed a sensitivity of 33% (95% CI 22% to 47%) and specificity of 98% (95% CI 96% to 99%). The clinical prediction rule showed a sensitivity of 40% (95% CI 27% to 54%) and specificity of 92% (95% CI 87% to 95%). Clinician judgment when patients presented within 48 hours showed a sensitivity of 67% (95% CI 39% to 86%) and specificity of 96% (95% CI 81% to 99%). Neither the rapid influenza test (P=.10) nor the clinical prediction rule (P=.42) was superior to clinician judgment alone in the diagnosis of influenza. CONCLUSION: The suggestion that a clinical decision rule or a rapid influenza test is better than clinical judgment alone for the diagnosis of influenza in an unselected patient population is not supported by this study. PMID- 16271672 TI - Update on emerging infections: news from the Centers for Disease Control and Prevention. Travel-associated dengue infections--United States, 2001-2004. PMID- 16271674 TI - Analysis of current management of atrial fibrillation in the acute setting: GEFAUR-1 study. AB - STUDY OBJECTIVE: Limited information relative to the management of atrial fibrillation in the emergency department (ED) daily practice is available. This study evaluates current management of atrial fibrillation in this setting to identify areas for practice improvement. METHODS: This was a prospective multicenter observational study carried out in 12 EDs. Adults in whom atrial fibrillation was demonstrated in an ECG obtained in the ED were included. Clinical variables and atrial fibrillation management in the ED were prospectively collected by the treating physicians using a standardized questionnaire. Patients with rapid ventricular response (>100 beats/min) were considered eligible for rate control, and patients with recent-onset episodes (<48 hours) were eligible for rhythm control. RESULTS: Of 1,178 patients, 41% presented with a rapid ventricular response and 21% had recent-onset episodes. Rhythm control was attempted in 42% of eligible patients, with antiarrhythmic drugs in 88% of cases (I-C drugs in 44% of patients; amiodarone in 43% of patients). Overall effectiveness of pharmacologic cardioversion was 63% (amiodarone 54.5%, flecainide 93%), whereas electrocardioversion was effective in 87.5% of cases. Rate control was performed in 68.3% of eligible patients (overall effectiveness 47.8%); digoxin was used in 67% of cases (effectiveness 45%). Both strategies were selected in 4.5% of cases, whereas no treatment for atrial fibrillation was performed in 60% of patients. CONCLUSION: In our ED population, rate-control effectiveness is poor and rhythm control is not attempted in most recent-onset episodes. Methods to improve rate-control effectiveness, the selection of patients for rhythm control, and the use of electrocardioversion appear warranted. PMID- 16271675 TI - Artificial neural network models for prediction of acute coronary syndromes using clinical data from the time of presentation. AB - STUDY OBJECTIVE: Clinical and ECG data from presentation are highly discriminatory for diagnosis of acute coronary syndromes, whereas definitive diagnosis from serial ECG and cardiac marker protein measurements is usually not available for several hours. Artificial neural networks are computer programs adept at pattern recognition tasks and have been used to analyze data from chest pain patients with a view to developing diagnostic algorithms that might improve triage practices in the emergency department. The aim of this study is to develop and optimize artificial neural network models for diagnosis of acute coronary syndrome, to test these models on data collected prospectively from different centers, and to establish whether the performance of these models was superior to that of models derived using a standard statistical technique, logistic regression. METHODS: The study used data from 3,147 patients presenting to 3 hospitals with acute chest pain. Data from hospital 1 were used to train the models, which were then tested on independent data from the other 2 hospitals. From 40 potential factors, variables were selected according to the logarithm of their likelihood ratios to produce models using 8, 13, 20, and 40 factors. Identical data were used for logistic regression and artificial neural network models. Calibration and performance were assessed, the latter using receiver operating characteristic (ROC) curve analysis. RESULTS: Although the performance of artificial neural network models generally increased with increasing numbers of factors, this was insignificant. The 13-factor model was therefore used for the rest of the study owing to its marginally improved calibration over the smallest model. Area under the ROC curve (with standard error) was 0.97 (0.006). The overall sensitivity and specificity of this model for acute coronary syndrome diagnosis using the training data was 0.93. ROC curves for logistic regression and artificial neural network models applied to data from the 3 hospitals were identical. For the 13-factor artificial neural network model tested on data from hospitals 2 and 3, area under the ROC curves (standard error) were 0.93 (0.006) and 0.95 (0.009), respectively. Investigation of the performance of the artificial neural network models throughout the range of predicted probabilities showed that they were well calibrated. CONCLUSION: This study confirms that artificial neural networks can offer a useful approach for developing diagnostic algorithms for chest pain patients; however, the exceptional performance and simplicity of the logistic model militates in favor of logistic regression for the present task. Our artificial neural network models were well calibrated and performed well on unseen data from different centers. These issues have not been addressed in previous studies. However, and unlike in previous studies, we did not find the performance of artificial neural network models to be significantly different from that of suitably optimized logistic regression models. PMID- 16271676 TI - Classification of emergency department chief complaints into 7 syndromes: a retrospective analysis of 527,228 patients. AB - STUDY OBJECTIVE: Electronic surveillance systems often monitor triage chief complaints in hopes of detecting an outbreak earlier than can be accomplished with traditional reporting methods. We measured the accuracy of a Bayesian chief complaint classifier called CoCo that assigns patients 1 of 7 syndromic categories (respiratory, botulinic, gastrointestinal, neurologic, rash, constitutional, or hemorrhagic) based on free-text triage chief complaints. METHODS: We compared CoCo's classifications with criterion syndromic classification based on International Classification of Diseases, Ninth Revision (ICD-9) discharge diagnoses. We assigned the criterion classification to a patient based on whether the patient's primary diagnosis was a member of a set of ICD-9 codes associated with CoCo's 7 syndromes. We tested CoCo's performance on a set of 527,228 chief complaints from patients registered at the University of Pittsburgh Medical Center emergency department (ED) between 1990 and 2003. We performed a sensitivity analysis by varying the ICD-9 codes in the criterion standard. We also tested CoCo on chief complaints from EDs in a second location (Utah). RESULTS: Approximately 16% (85,569/527,228) of the patients were classified according to the criterion standard into 1 of the 7 syndromes. CoCo's classification performance (number of cases by criterion standard, sensitivity [95% confidence interval (CI)], and specificity [95% CI]) was respiratory (34,916, 63.1 [62.6 to 63.6], 94.3 [94.3 to 94.4]); botulinic (1,961, 30.1 [28.2 to 32.2], 99.3 [99.3 to 99.3]); gastrointestinal (20,431, 69.0 [68.4 to 69.6], 95.6 [95.6 to 95.7]); neurologic (7,393, 67.6 [66.6 to 68.7], 92.7 [92.6 to 92.8]); rash (2,232, 46.8 [44.8 to 48.9], 99.3 [99.3 to 99.3]); constitutional (10,603, 45.8 [44.9 to 46.8], 96.6 [96.6 to 96.7]); and hemorrhagic (8,033, 75.2 [74.3 to 76.2], 98.5 [98.4 to 98.5]). The sensitivity analysis showed that the results were not affected by the choice of ICD-9 codes in the criterion standard. Classification accuracy did not differ on chief complaints from the second location. CONCLUSION: Our results suggest that, for most syndromes, our chief complaint classification system can identify about half of the patients with relevant syndromic presentations, with specificities higher than 90% and positive predictive values ranging from 12% to 44%. PMID- 16271677 TI - Ultrasonography-guided peripheral intravenous access versus traditional approaches in patients with difficult intravenous access. AB - STUDY OBJECTIVE: We assess the success rate of emergency physicians in placing peripheral intravenous catheters in difficult-access patients who were unsuccessfully cannulated by emergency nurses. A technique using real-time ultrasonographic guidance by 2 physicians was compared with traditional approaches using palpation and landmark guidance. METHODS: This was a prospective, systematically allocated study of all patients requiring intravenous access who presented to 2 university hospitals between October 2003 and March 2004. Inclusion criterion was the inability of any available nurse to obtain intravenous access after at least 3 attempts on a subgroup of patients who had a history of difficult intravenous access because of obesity, history of intravenous drug abuse, or chronic medical problems. Exclusion criterion was the need for central venous access. Patients presenting on odd days were allocated to the ultrasonographic-guided group, and those presenting on even days were allocated to the traditional-approach group. Endpoints were successful cannulation, number of sticks, time, and patient satisfaction. RESULTS: Sixty patients were enrolled, 39 on odd days and 21 on even days. Success rate was greater for the ultrasonographic group (97%) versus control (33%), difference in proportions of 64% (95% confidence interval [CI] 39% to 71%). The ultrasonographic group required less overall time (13 minutes versus 30 minutes, for a difference of 17 [95% CI 0.8 to 25.6]), less time to successful cannulation from first percutaneous puncture (4 minutes versus 15 minutes, for a difference of 11 [95% CI 8.2 to 19.4]), and fewer percutaneous punctures (1.7 versus 3.7, for a difference of 2.0 [95% CI 1.27 to 2.82]) and had greater patient satisfaction (8.7 versus 5.7, for a difference of 3.0 [95% CI 1.82 to 4.29]) than the traditional landmark approach. CONCLUSION: Ultrasonographic-guided peripheral intravenous access is more successful than traditional "blind" techniques, requires less time, decreases the number of percutaneous punctures, and improves patient satisfaction in the subgroup of patients who have difficult intravenous access. PMID- 16271679 TI - Images in emergency medicine. Synypnea. PMID- 16271680 TI - Images in emergency medicine. Human ehrlichiosis. PMID- 16271681 TI - The costs of visits to emergency departments--revisited. PMID- 16271683 TI - Rectal insertion of fentanyl patches: a new route of toxicity. PMID- 16271686 TI - Sensitivity of the Patient-rated Forearm Evaluation Questionnaire in lateral epicondylitis. AB - The purpose of this study was to examine the sensitivity, reliability, and concurrent validity of the Patient-rated Forearm Evaluation Questionnaire (PRFEQ). Reliability on three consecutive days was evaluated with 22 of 94 subjects who had chronic lateral epicondylitis (LE) and who concomitantly participated in an outcome study. The PRFEQ results were compared with results of the Visual Analogue Scale; the Disabilities of the Arm, Shoulder, and Hand questionnaire; the Medical Outcomes Study 36-Item Short Form Health Survey; and the pain-free grip strength measurement. Questionnaires were completed at baseline, six weeks, and 12 weeks. Reliability was excellent using variance components and interclass correlation coefficients (PRFEQ function subscale, 0.92; PRFEQ pain subscale, 0.96; PRFEQ total scale, 0.96). Generally, correlations were moderate between the PRFEQ subscales and total scale and the other outcome scales. Effect size and standardized response mean were good in many outcome scales, being slightly higher in the PRFEQ than in the other outcome measures. The PRFEQ is reliable, reproducible, and sensitive for assessment of LE. It is at least as sensitive to change as the other outcome tools tested. The PRFEQ should be a standard primary outcome measure in LE research. PMID- 16271687 TI - Update: The Patient-rated Forearm Evaluation Questionnaire is now the Patient rated Tennis Elbow Evaluation. PMID- 16271688 TI - Chronic lateral epicondylitis: comparative effectiveness of a home exercise program including stretching alone versus stretching supplemented with eccentric or concentric strengthening. AB - The objective of this study was to evaluate the effectiveness of eccentric strengthening. Ninety-four subjects (50 men) with chronic lateral epicondylitis were allocated randomly into three groups: stretching, concentric strengthening with stretching, and eccentric strengthening with stretching. Subjects performed an exercise program for six weeks. All three groups received instruction on icing, stretching, and avoidance of aggravating activities. The strengthening groups received instruction on isolated concentric and eccentric wrist extensor strengthening, respectively. At six weeks, significant gains were made in all three groups as assessed with pain-free grip strength, Patient-rated Forearm Evaluation Questionnaire, Disabilities of the Arm, Shoulder, and Hand questionnaire, Short Form 36, and visual analog pain scale. No significant differences in outcome measures were noted among the three groups. Although there were no significant differences in outcome among the groups, eccentric strengthening did not cause subjects to worsen. Further studies are needed to assess the unique effects of a more intense or longer eccentric strengthening program for patients with lateral epicondylitis. PMID- 16271689 TI - The influence of age on pressure perception of static and moving two-point discrimination in normal subjects. AB - The purpose of the present study was to determine the effect of age on digital pressure perception as measured by two-point discrimination (2PD) testing. The subjects were 177 normal volunteers ranging in age from 20 to 79 years. Perceptible pressure of static and moving 2PD was measured on the index finger and little finger, using the Pressure-specifying Sensory Device. The threshold of pressure perception increased significantly with advancing age in both static and moving 2PD tests. There was a marked increase in subjects older than 60 years. Pressure perception was significantly higher for static 2PD than for moving 2PD in subjects 70-79 years of age. The threshold of pressure perception for static and moving 2PD gradually increased with advancing age, and was markedly elevated in subjects older than 60 years. PMID- 16271690 TI - Test-retest reliability of grip-strength measures obtained over a 12-week interval from community-dwelling elders. AB - This study examined the test-retest reliability of hand-grip strength measured over a 12-week period. Twenty-one apparently healthy elders (age=75.0+/-5.9 yr) participated. Single strength measurements obtained bilaterally using a Jamar dynamometer were compared across time. Test and retest measurements did not differ significantly over time on either side. Intraclass correlation coefficients were 0.954 and 0.912 for the left and right hands, respectively. Technical errors of measurement were 15.8 and 21.3N for the left and right hands, respectively. Measurements of hand-grip strength obtained from elders over a 12 week period are reliable. PMID- 16271691 TI - Contrasting perspectives on pain following hand injury. AB - The purposes of this article are 1) to compare contrasting ways of documenting pain, 2) to identify the usefulness of information each method provides, and 3) to examine implications for hand therapy. Participants are tracked for 12 months in an ongoing study of Adaptation to Hand Injury that incorporates quantitative measures of both intensity and functional impact of pain, as well as semistructured qualitative Adaptation Interviews that elicit and document patients' responses to pain and injury experience. Each method of documenting pain tells us something important. Numeric measures of pain intensity (Visual Analog Scale) allow us to document change over time and identify relationships between intensity and self-reported changes in daily life as documented in a structured questionnaire (Disabilities of Arm, Shoulder, and Hand). Qualitative interviews provide an individual perspective on responses to pain and disability and the personal beliefs and experiences that influenced the responses of each unique individual. Findings support the importance of studying more about the clinical behaviors of our patients for better treatment outcomes and recommend the use of qualitative research in helping us to uncover this knowledge. PMID- 16271692 TI - Patients' adaptive experiences of returning to work following musculoskeletal disorders: a mixed design study. AB - Compelling evidence from multiple studies has identified possible causes of upper extremity musculoskeletal disorders (MSDs). MSDs are multifactorial in origin with a number of risk factors contributing to their development. Despite studies attempting to provide change and prevention, the rise in MSD incidence has not slowed. To fully understand the etiology, it is important to consider the entire spectrum of possible causes. The purpose of this paper is to explore the experiences of persons with upper extremity musculoskeletal disease and their adaptation back to work. A mixed design explorative study involving quantitative questionnaires and qualitative interviews with MSD subjects was conducted. The methodology included completion of the Disability of Arm, Shoulder, and Hand questionnaire (DASH), Work Environment Scale (WES), and interviews. DASH results demonstrated meaningful changes in the subscales "overall function" and "function at work." The WES showed positive associations in all subscales. Comparison across the cases during interviews revealed four major themes characterizing the adaptive process. PMID- 16271693 TI - Static progressive splint for composite flexion. AB - Frequently when one sustains an injury to a digit, multiple soft tissue structures can be involved. As a result, the MCP, PIP, and DIP joints can be affected, thus limiting the ability to fully flex the finger. As hand therapists, one of our primary goals is to improve functional grasp/grip. These authors have designed a single-digit, composite flexion splint to help achieve this goal. PMID- 16271694 TI - Lipoprotein profiles and components in Tsaiya ducks under ad libitum feeding and fasting. AB - This study investigated lipoprotein compositions and distribution in Tsaiya ducks under ad libitum feeding and fasting. The study was conducted on each of thirty Tsaiya ducks over two 4-week periods. Each of the 30 Tsaiya ducks was randomly allocated into an ad libitum feeding group or a 3-day fasting group during the growing period (8-12 weeks old) and during the initial laying period (26-30 weeks old; 10-14 weeks after the onset of laying), respectively. Experimental results indicated that HDL (high-density lipoprotein) is the primary lipoprotein species in growing ducks but that in laying ducks the major population of lipoprotein is VLDL (very low density lipoprotein). Protein and cholesteryl ester were identified as the major components of HDL during both periods; triacylglycerol (TG) was the largest proportion of VLDL in both periods. In laying ducks VLDL contains more TG than in growing ducks; VLDL particle sizes in laying ducks were larger than in growing ducks. Fasting significantly decreased VLDL levels during both periods. The TG content in most lipoproteins was also decreased by fasting during both test periods. In contrast, the cholesteryl ester and free cholesterol in most lipoproteins were increased by fasting. The particle size of HDL and VLDL also decreased during fasting. This study indicated that lipoprotein profiles and components in Tsaiya ducks during the laying period were different from during the growing period and were affected by fasting. PMID- 16271695 TI - The validation of a new actigraphy system for the measurement of periodic leg movements in sleep. AB - BACKGROUND AND PURPOSE: To determine the accuracy of a new actigraphy system for the measurement of periodic leg movements (PLMs). Continuous measurements of leg movements, made overnight, are essential to diagnose and monitor treatment for PLMs. We have developed the Actiwatch with Cambridge Neuro-Technology Ltd (Cambridge, UK) to detect leg movements consistent with PLMs, to record these movements from both feet over three consecutive nights and to report standard indices of PLMs. We describe three studies designed to validate this device. PATIENTS AND METHODS: The Actiwatch was assessed on a bench model across a range of movement amplitudes. The level of agreement on individual movements between the Actiwatch and bilateral anterior tibialis electromyography (BATEMG) measures was assessed in 199 epochs from five patients with known PLMs. The ability of the Actiwatch to correctly identify patients with PLMs was assessed in a comparison with polysomnography (PSG) in 50 consecutive patients investigated in the sleep laboratory. RESULTS: The Actiwatch detected all mechanically generated movements (100% sensitivity and 100% specificity) on the bench test. On individual movements BATEMG measurement and the Actiwatch agreed on 94% of epochs. The sensitivity and specificity of the Actiwatch to detect leg movements in severe PLMs as defined by BATEMG (PLMI>or=25) are high (100% sensitive and 97% specific). Our bilateral system gave better agreement with BATEMG than previous systems but the two measures are still not interchangeable. CONCLUSIONS: EMG is only a surrogate measure of limb movement and indices measured in this way correlate poorly with symptoms. We have demonstrated that the Actiwatch faithfully records movement, which offers the potential to reassess the relationships between limb movement indices and symptoms. This which offers a convenient and economical alternative to PSG in the study of large populations to increase our understanding of the epidemiology and clinical significance of PLMs. PMID- 16271696 TI - Prevalence and determinants of excessive daytime sleepiness in an Asian multi ethnic population. AB - BACKGROUND AND PURPOSE: To investigate the prevalence and determinants of daytime sleepiness in an Asian multi-ethnic population. Cross-sectional survey based on a questionnaire in multi-ethnic Asian adult population of Singapore (2.4 million). PATIENTS AND METHODS: Measured frequency of excessive daytime sleepiness (EDS), socio-demographic status, sleep duration, perceived sleep insufficiency, frequent awakenings, difficulty falling asleep, snoring, breathing pauses, GHQ-12, shift work, solvent exposure at work, substance use, medications, and chronic diseases in random sample of 2,298 adults, aged 20-74 years, representing Chinese, Malays and Indians. RESULTS: Among the respondents, 10.8% reported usually feeling sleepy in the daytime, and 9.0% were classified as having EDS. Marked ethnic difference was observed, with higher rates of EDS in Malays and Chinese, compared to Indians. Increased risk of EDS was associated with perceived sleep insufficiency; snoring and breathing pauses; shift work and solvent exposure at work; psychiatric morbidity and chronic diseases, particularly chronic rhinitis; and medications with sedating effects. CONCLUSIONS: Daytime sleepiness is associated with a multiplicity of highly prevalent risk factors in the community, including sleep behavior, breathing-, work- and medically related factors. Significant ethnic differences, which were not explained by these factors, were observed between Chinese, Malays and Indians. PMID- 16271698 TI - Probing the sterol binding site of soybean sterol methyltransferase by site directed mutagenesis: functional analysis of conserved aromatic amino acids in Region 1. AB - Soybean sterol methyltransferase (SMT) in the presence of AdoMet catalyzes the transmethylation of the delta24-bond of the sterol side chain to produce phytosterols with a methyl(lene) or ethyl(idene) group at C-24. The function of six aromatic amino acids associated with the putative active center of the SMT, i.e., Region 1 that extends from Phe82 to Phe93 in soybean SMT, was studied by site-directed mutagenesis and heterologous expression in BL21(DE3) bacterial cells. The enzyme-generated products were characterized kinetically and by GC-MS analysis. Substitution of the aromatic amino acids at positions 82, 83, 85, 87, 91, and 93 with a leucine residue produced mutant SMTs with varying activities. The mutants converted cycloartenol to 24(28)-methylene cycloartanol [C1-activity] from a few percent to as much as 95% of the control activity. In contrast, none of the leucine mutants were found to catalyze 24(28)-methylene lophenol [C2 activity], suggesting a loss of function associated with the second C1-transfer activity. In contrast to the loss of the second C1-transfer activity of the Phe82Leu, replacement of the Phe82 residue to isoleucine had minimal effect on the first or second C1-transfer activities, suggesting that the increased bulk (branching) in the leucine side chain contributes to significant perturbations in the active site that generate inaccurate positioning of the substrate side chain disfavoring the second C1-transfer activity. Replacement of Tyr83 to phenylalanine resulted in an increase of the specificity constant (kcat/Km) for the substrate of the second C1-transfer activity by a factor of 5 compared to control and an increase of delta24(28)Z-ethylidene sterol formation in the 24 ethyl sterol product set, suggesting that loss of steric bulk from the phenolic hydroxyl group on tyrosine generates a less precise fit of the delta24(28) sterol side chain into the active site favoring the second C1-transfer activity and prompting reaction channeling during catalysis. Circular dichroism spectra, equilibrium dialysis studies of AdoMet, and chromatographic information of the wild-type and Tyr83 mutants confirmed retention of the overall conformation of the enzyme during the experiments. Together, these findings suggest that the amino acids of Region 1 provide a tight substrate orientation imposed by hydrophobic interactions between the sterol side chain and the SMT active site contacts and control the production and processing of the transmethylation pathways governed by the first and second C1-transfer activities. PMID- 16271699 TI - Immunohistochemical expression of keratins 13 and 14 in the lingual epithelium of rats during the morphogenesis of filiform papillae. AB - We examined the immunofluorescence of keratins 13 (K13) and 14 (K14) and differential interference contrast (DIC) images during the morphogenesis of filiform papillae and the keratinization of the lingual epithelium of rats on semi-ultrathin sections of epoxy resin-embedded samples by laser-scanning microscopy. We also examined semi-ultrathin sections of epoxy resin embedded, toluidine blue stained samples by light microscopy to obtain details of cell histology and morphology. No immunoreactivity specific for K13 and K14 was detected on the lingual epithelium of foetuses on days 13, 15 and 17 after conception (E13, E15 and E17), during which time the number of layers of cuboidal cells in the lingual epithelium increased from one to several. Immunoreactivity specific for K13 and K14 was first detected on the lingual epithelium of foetuses on E19. The immunoreactivity specific for K13 appeared in the suprabasal cells of the papillary and interpapillary cell columns and immunoreactivity specific for K14 was detected in the basal and suprabasal cells of the papillary and interpapillary cell columns. The lingual epithelium was composed of stratified squamous cells. The rudiments of filiform papillae were compactly arranged and interpapillary cell columns were very narrow. Filiform papillae developed gradually from postnatal day 0 (PO) to 21 (P21). The width of interpapillary spaces also increased during this period. Immunoreactivity specific for K13 and K14 was distinct at all postnatal stages examined. Thus, the patterns of immunoreactivity of K13 and K14 differed as the filiform papillae developed. PMID- 16271700 TI - Multiple Rieske proteins in prokaryotes: where and why? AB - Many microbial genomes have been sequenced in the recent years. Multiple genes encoding Rieske iron-sulfur proteins, which are subunits of cytochrome bc-type complexes or oxygenases, have been detected in many pro- and eukaryotic genomes. The diversity of substrates, co-substrates and reactions offers obvious explanations for the diversity of the low potential Rieske proteins associated with oxygenases, but the physiological significance of the multiple genes encoding high potential Rieske proteins associated with the cytochrome bc-type complexes remains elusive. For some organisms, investigations into the function of the later group of genes have been initiated. Here, we summarize recent finding on the characteristics and physiological functions of multiple high potential Rieske proteins in prokaryotes. We suggest that the existence of multiple high potential Rieske proteins in prokaryotes could be one way of allowing an organism to adapt their electron transfer chains to changing environmental conditions. PMID- 16271701 TI - In vivo plasmid DNA electroporation resulted in transfection of satellite cells and lasting transgene expression in regenerated muscle fibers. AB - In vivo plasmid DNA electroporation resulted in elevated and lasting transgene expression in skeletal muscles. But the nature of the cells that contributed to sustained gene expression remains unknown. We followed the fate of plasmid DNA delivered with electroporation and systematically investigated the time course and location of transgene expression in muscle tissues both with GFP and luciferase. Furthermore, satellite cell activation after electroporation was confirmed by RT-PCR and immunohistochemistry analysis. The activated satellite cells were shown to be able to uptake the injected plasmid DNA and express transgene products as regenerated myocytes. We found that cells with longer gene expression durations were mostly regenerated muscle fibers. In contrast, expression in pre-existing muscle fibers was rather transient. We also presented in this study that immune response to transgene products might hamper the lasting gene expression. Based on these observations, we proposed that the underlying mechanism for prolonged transgene expression in the muscles after electroporation is related to the activation and transfection of myogenic satellite cells which subsequently developed into regenerated muscle fibers. PMID- 16271702 TI - BIP co-chaperone MTJ1/ERDJ1 interacts with inter-alpha-trypsin inhibitor heavy chain 4. AB - MTJ1/ERdj1 and its human homologue HTJ1 are membrane proteins that interact with the molecular chaperone BiP through their J-domain. HTJ1 also contains a C terminal cytosolic region of unknown function that consists of two SANT domains separated by a spacer region. We recently showed that the second SANT domain of HTJ1 (SANT2) binds to alpha1-antichymotrypsin and alters its serpin activity [B. Kroczynska, C.M. Evangelista, S.S. Samant, E.C. Elguindi, S.Y. Blond, The SANT2 domain of the murine tumor cell DnaJ-like protein 1 human homologue interacts with alpha1-antichymotrypsin and kinetically interferes with its serpin inhibitory activity, J. Biol. Chem. 279 (2004) 11432-11443]. Here, we identified a new variant of human inter-alpha-trypsin inhibitor heavy chain 4 (ITIH4) that also interacts with the SANT2 domain of HTJ1. Biochemical, mutagenesis, and fluorescence studies demonstrate that SANT2 binds to a carboxyl-terminal fragment that corresponds to the last third of the new ITIH4 isoform sequence (residues 588-930). ITIH4 and MTJ1 co-immunoprecipitate from total liver protein extracts and SANT2 protects the ITIH4(588-930) recombinant fragment from being processed by kallikrein in vitro. This work reveals that the SANT2 domain of HTJ1 is a genuine protein-protein interaction module. PMID- 16271703 TI - A role of opening of mitochondrial ATP-sensitive potassium channels in the infarct size-limiting effect of ischemic preconditioning via activation of protein kinase C in the canine heart. AB - The opening of mitochondrial ATP-sensitive K+ (mitoK(ATP)) channels triggers or mediates the infarct size (IS)-limiting effect of ischemic preconditioning (IP). Because ecto-5'-nucleotidase related to IP is activated by PKC, we tested whether the opening of mitoK(ATP) channels activates PKC and contributes to either activation of ecto-5'-nucleotidase or IS-limiting effect. In dogs, IP procedure decreased IS and activated ecto-5'-nucleotidase, both of which were mimicked by transient exposure to either cromakalim or diazoxide, and these effects were blunted by either GF109203X (a PKC inhibitor) or 5-hydroxydecanoate (a mitoK(ATP) channel blocker), but not by HMR-1098 (a surface sarcolenmal K(ATP) channel blocker). Either cromakalim or diazoxide activated both PKC and ecto-5' nucleotidase, which was blunted by either GF109203X or 5-hydroxydecanoate, but not by HMR-1098. We concluded that the opening of mitoK(ATP) channels contributes to either activation of ecto-5'-nucleotidase or the infarct size-limiting effect via activation of PKC in canine hearts. PMID- 16271705 TI - Denaturing HPLC-based approach for detection of COL7A1 gene mutations causing dystrophic epidermolysis bullosa. AB - Dystrophic epidermolysis bullosa (DEB) is a rare clinically heterogeneous genodermatosis due to genetic defects in type VII collagen gene (COL7A1). Identification of COL7A1 mutations is a challenge since this gene comprises 118 exons and more than 300 mutations scattered over the gene have been reported. Here, we describe for the first time the use of denaturing high performance liquid chromatography (DHPLC) for COL7A1 mutation detection. To validate the method, exon-specific DHPLC conditions were applied to screen DNA samples from patients carrying known COL7A1 mutations. Abnormal DHPLC profiles were obtained for all known mutations. Subsequent DHPLC analysis of 17 DEB families of unknown genotype allowed the identification of 21 distinct mutations, 9 of which were novel. The DHPLC mutation detection rate was significantly higher compared with our mutation scanning rate with conventional techniques (97% vs 86%), indicating DHPLC as the method of choice for COL7A1 molecular characterization in DEB patients. PMID- 16271704 TI - Simvastatin induces impairment in skeletal muscle while heart is protected. AB - 3-Hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors (statins) are widely used to reduce plasma cholesterol concentration. However, statins are also known to induce various forms of muscular toxicity. We have previously shown that acute application of simvastatin on human skeletal muscle samples induced a cascade of cellular events originating from mitochondria and resulting in a global alteration of Ca2+ homeostasis. The present study was designed to further define the origin of the mitochondria impairment and to understand the apparent lack of deleterious effect on the heart. Using fluorescence imaging analysis and oxygraphy on human and rat skinned skeletal muscle samples, we show that the simvastatin-induced mitochondria impairment results from inhibition of the complex I of respiratory chain. Similar simvastatin-induced mitochondria impairment and alteration of Ca2+ homeostasis occur in permeabilized but not in intact ventricular rat cardiomyocytes. In intact rat skeletal muscle fibers from the flexor digitorum brevis muscle, the simvastatin-induced alteration of Ca2+ homeostasis is abolished when monocarboxylate transporter (MCT4) is inhibited. The impairment of complex I by simvastatin might be the primary step of its cellular deleterious effects leading to muscle fiber death. This mechanism is seen specifically in skeletal muscles. This specificity should be in part attributed to a preferential uptake of statins by MCT4 that is not expressed in cardiomyocytes. PMID- 16271706 TI - Hypophosphoric acid is a unique substrate of pyrophosphorolysis catalyzed by HIV 1 reverse transcriptase. AB - Pyrophosphate analogues, namely, pyrophosphorous, hypophosphoric, and hypophosphorous acids, were evaluated as inhibitors in elongation reactions and substrates in pyrophosphorolysis reaction catalyzed by HIV-1 reverse transcriptase and DNA polymerase I (the Klenow fragment). The substrate efficacy of hypophosphoric acid in pyrophosphorolysis reaction exceeded that of pyrophosphate for both enzymes by more than ten times. The product of the reaction was a dNTP analogue bearing a hypophosphate in the beta,gamma-position. Pyrophosphorous and hypophosphorous acids were neither inhibitors nor substrates for the enzymes. Kinetic parameters of the pyrophosphorolysis reaction catalyzed by HIV reverse transcriptase in the presence of hypophosphoric acid were evaluated. The dTMP analogue bearing a hypophosphate in the beta,gamma-position was synthesized and its substrate properties in elongation reaction catalyzed by HIV-1 reverse transcriptase were similar to those of natural dTTP. Hypophosphoric acid was capable of removing ddTMP, ddTMP(3'N3), and ddTMP(3'NH2) from the 3'-end of primers with an equal efficacy. PMID- 16271707 TI - Differential interactions of G-proteins and adenylyl cyclase with nucleoside 5' triphosphates, nucleoside 5'-[gamma-thio]triphosphates and nucleoside 5' [beta,gamma-imido]triphosphates. AB - The regulatory G-proteins of adenylyl cyclase (AC), G(i) and G(s), are not only activated by GTP and the stable GTP analogs, guanosine 5'-[gamma thio]triphosphate (GTPgammaS) and guanosine 5'-[beta,gamma-imido]triphosphate (GppNHp), but also by hypoxanthine, xanthine, uracil and cytidine nucleotides. The latter nucleotides were previously used to analyze distinct active G-protein states. Surprisingly, recent studies have shown that inosine 5'-[gamma thio]triphosphate and uridine 5'-[gamma-thio]triphosphate can also inhibit AC directly. Therefore, we systematically compared the interactions of nucleoside 5' triphosphates (NTPs), nucleoside 5'-[gamma-thio]triphosphates (NTPgammaSs) and nucleoside 5'-[beta,gamma-imido]triphosphates (NppNHps) with G(i), G(s) and AC. NTPgammaSs exhibited up to 26,000-fold higher affinity for G-proteins than NTPs and NppNHps. NTPgammaSs were up to 150-fold more potent direct AC inhibitors than NTPs and NppNHps. G-proteins exhibited striking preference for guanine nucleotides compared to other purine and pyrimidine nucleotides, whereas base selectivity of various ACs, particularly the purified catalytic subunits C1.C2, was rather poor. GTP, GTPgammaS and GppNHp exhibited much higher selectivity for G-proteins relative to AC than all other purine and pyrimidine nucleotides. We have energetically characterized the interactions of purine and pyrimidine nucleotides with AC in silico, constructing pharmacophore models that correlate well with experimental affinities and have elucidated specific amino acid residues with greatest influence on nucleotide binding. Collectively, both G proteins and ACs bind purine and pyrimidine nucleotides, with G-proteins showing much higher base-selectivity than AC. Thus, direct inhibitory effects of nucleotides on AC should be understood and considered when probing distinct active G-protein states with non-guanine nucleotides. PMID- 16271708 TI - Molecular cloning of Clock cDNA from the prawn, Macrobrachium rosenbergii. AB - CLOCK, which belongs to the basic helix-loop-helix (bHLH)/PER-ARNT-SIM (PAS) superfamily of transcription factors, is one of the most essential proteins involved in circadian systems of animals. Clock genes have been cloned from several species, including mammals, insects, birds, fish, and amphibians. In the present study, we successfully isolated a Clock homolog (termed Mar-Clock) from the giant prawn, Macrobrachium rosenbergii. The 2949-bp cDNA contained a 2115 bp open reading frame that encoded a putative CLOCK protein of 704 amino acids (termed Mar-CLOCK) exhibiting high identities with CLOCK homologs in other species (30-35%). This is the first report of a circadian clock gene from crustaceans. Mar-CLOCK possessed an exceptionally long glutamine-rich domain (140 amino acids) in its C-terminus, which usually ranges from 14 to 57 amino acids in other known CLOCKs and is supposed to function in transcriptional activation. Using RT-PCR, we observed that Mar-Clock was expressed in all tested tissues. Semiquantitative RT-PCR was performed to investigate the gene expression profile during the light-dark cycle. The results indicated that the expression of the Mar Clock gene had no significant rhythmicity in central nervous tissues (thoracic ganglia and eyestalk) or peripheral tissues (gill, ovary, hepatopancreas, and muscle). Furthermore, gene expression tended to increase in the central nervous system (brain, thoracic, and abdominal ganglia) of eyestalk-ablated or constant dark (DD) prawns, and in the eyestalk-ablated gill. No expression change was found under constant light (LL) or in heart and muscle. PMID- 16271709 TI - Quetiapine reverses the suppression of hippocampal neurogenesis caused by repeated restraint stress. AB - Quetiapine is an atypical antipsychotic effective in treating the positive, negative, and cognitive symptoms of patients with schizophrenia. Our previous study has shown that chronic administration of quetiapine attenuates the decrease in levels of brain-derived neurotrophic factor (BDNF) in the hippocampi of rats subjected to chronic-restraint stress. In the present study, we investigated the effects of quetiapine on hippocampal neurogenesis that had been compromised in stressed rats. Newborn cells in the hippocampus were labeled by bromodeoxyuridine (BrdU), and immature neurons were detected immunohistochemically using an antibody against phosphorylated cAMP response element-binding protein (pCREB). The restrained rats (4 h/day for 7 days) showed lower levels of hippocampal neurogenesis indicated by decreased numbers of BrdU-labeled and pCREB-positive cells. Post-stress administration of quetiapine (10 mg/kg) for 7 or 21 days reversed the stress-induced suppression of hippocampal neurogenesis, evidenced in the numbers of BrdU-labeled and pCREB-positive cells that are comparable to those in non-stressed rats but higher than those in the vehicle-treated rats. The results may help us understand the therapeutic effects of quetiapine on cognitive deficits in patients with schizophrenia and depression, in which the structure and functions of the hippocampus are implicated. PMID- 16271710 TI - Resistin is secreted from macrophages in atheromas and promotes atherosclerosis. AB - OBJECTIVE: Resistin belongs to a family of cysteine-rich secreted polypeptides that are mainly produced by monocytes/macrophages in humans. Recently, high concentrations of resistin were shown to induce vascular endothelial dysfunction and vascular smooth muscle cell proliferation. We examined if resistin was secreted from macrophages locally in atheromas and if it affected vascular cell function in human. METHODS AND RESULTS: Immunohistochemical staining of human vessels showed that aortic aneurysms exhibited resistin-positive staining areas along macrophage infiltration, while normal arteries and veins did not. Co localization of resistin and CD68 (a marker for macrophages) was observed in immunofluorescent double staining of aneurysms. Resistin mRNA was expressed much higher in cultured monocytes/macrophages than in human vascular smooth muscle cells (VSMCs) and human umbilical venous endothelial cells (HUVECs). This suggested that the resistin in aneurysms originates from macrophages within the vessels. To determine the effects of resistin on atherosclerosis, HUVECs and human VSMCs were incubated with resistin (10-100 ng/mL for 4 approximately 24 h). In HUVECs, plasminogen activator inhibitor (PAI)-1 release was assayed by ELISA, while the PAI-1 and endothelin (ET)-1 mRNA levels were analyzed by Northern blotting. Both were increased significantly with resistin treatment by factors of 1.3-2.5 (p<0.05). Migratory activity of VSMCs measured by scratched wound assay also increased significantly (1.6 times, p<0.05). In summary, macrophages infiltrating atherosclerotic aneurysms secrete resistin, and resistin affects endothelial function and VSMC migration. CONCLUSIONS: Resistin secreted from macrophages may contribute to atherogenesis by virtue of its effects on vascular endothelial cells and smooth muscle cells in humans. PMID- 16271711 TI - Use of genetic immunization to raise antibodies recognizing toxin-related cell surface ADP-ribosyltransferases in native conformation. AB - ADP-ribosyltransferases (ARTs) transfer ADP-ribose from NAD to arginine, asparagine, or cysteine residues in target proteins. This post-translational protein modification is the mechanism by which cholera-toxin and other bacterial toxins cause pathology in human host cells. Molecular cloning has identified five toxin-related GPI-anchored cell surface ARTs in the mouse (ART1, ART2.1, ART2.2, ART3, and ART4) and three in the human (ART1, ART3, and ART4). ART2-which has sparked interest because of its ability to activate the cytolytic P2X7 purinergic receptor by ADP-ribosylation-is encoded by two functional gene copies in the mouse genome while the human genome carries two inactivated ART2 pseudogenes. We generated stable transfectants for FLAG-tagged versions of each of the functional human and mouse ARTs. Using genetic immunization we raised monoclonal antibodies that recognize the native human ARTs on the surface of living cells. Some of these mAbs recognize an epitope shared with the mouse ART orthologue but not with more distant ART paralogues. Screening of primary cells and established cell lines by FACS revealed expression of ART1 by monocytes, neutrophils and myeloid leukemia cell lines but not by cell lines derived from solid tumors. ART1 and ART4 have been assigned the designations: CD296, and CD297, respectively. PMID- 16271712 TI - Relative contribution of cell contact pattern, specific PKC isoforms and gap junctional communication in tight junction assembly in the mouse early embryo. AB - In mouse early development, cell contact patterns regulate the spatial organization and segregation of inner cell mass (ICM) and trophectoderm epithelium (TE) during blastocyst morphogenesis. Progressive membrane assembly of tight junctional (TJ) proteins in the differentiating TE during cleavage is upregulated by cell contact asymmetry (outside position) and suppressed within the ICM by cell contact symmetry (inside position). This is reversible, and immunosurgical isolation of the ICM induces upregulation of TJ assembly in a sequence that broadly mimics that occurring during blastocyst formation. The mechanism relating cell contact pattern and TJ assembly was investigated in the ICM model with respect to PKC-mediated signaling and gap junctional communication. Our results indicate that complete cell contact asymmetry is required for TJ biogenesis and acts upstream of PKC-mediated signaling. Specific inhibition of two PKC isoforms, PKCdelta and zeta, revealed that both PKC activities are required for membrane assembly of ZO-2 TJ protein, while only PKCzeta activity is involved in regulating ZO-1alpha+ membrane assembly, suggesting different mechanisms for individual TJ proteins. Gap junctional communication had no apparent influence on either TJ formation or PKC signaling but was itself affected by changes of cell contact patterns. Our data suggest that the dynamics of cell contact patterns coordinate the spatial organization of TJ formation via specific PKC signaling pathways during blastocyst biogenesis. PMID- 16271714 TI - Genetic mechanisms of fetal male undermasculinization: a background to the role of endocrine disruptors. AB - Fetal male sex development proceeds along an orderly sequence of events coordinated by an interplay of genetic and hormonal events. These operate in a time- and concentration-dependent manner. Once a testis is formed (the female sex being constitutive in nature), differentiation of the internal and external male genitalia is androgen dependent. A number of genetic syndromes of sex reversal are well characterized at the biochemical and molecular levels. They fall into three principal categories: defects in formation of the testis, defects in production of androgens, and defects in the action of androgens. In many instances, the precise cause is not established, although the investigative evidence points in the direction of one of the three stated classifications. Polymorphic variants in several of the genes involved in male development are associated with certain degrees of male undermasculinization. While the genetic background is essentially static, it is plausible that the effect of endocrine disruptors during fetal life acting through epigenetic mechanisms may partly explain the observed changing trends in male reproductive tract disorders. PMID- 16271713 TI - Tissues and signals involved in the induction of placodal Six1 expression in Xenopus laevis. AB - Ectodermal placodes, from which many cranial sense organs and ganglia develop, arise from a common placodal primordium defined by Six1 expression. Here, we analyse placodal Six1 induction in Xenopus using microinjections and tissue grafts. We show that placodal Six1 induction occurs during neural plate and neural fold stages. Grafts of anterior neural plate but not grafts of cranial dorsolateral endomesoderm induce Six1 ectopically in belly ectoderm, suggesting that only the neural plate is sufficient for inducing Six1 in ectoderm. However, extirpation of either anterior neural plate or of cranial dorsolateral endomesoderm abolishes placodal Six1 expression indicating that both tissues are required for its induction. Elevating BMP-levels blocks placodal Six1 induction, whereas ectopic sources of BMP inhibitors expand placodal Six1 expression without inducing Six1 ectopically. This suggests that BMP inhibition is necessary but needs to cooperate with additional factors for Six1 induction. We show that FGF8, which is expressed in the anterior neural plate, can strongly induce ectopic Six1 in ventral ectoderm when combined with BMP inhibitors. In contrast, FGF8 knockdown abolishes placodal Six1 expression. This suggests that FGF8 is necessary and together with BMP inhibitors sufficient to induce placodal Six1 expression in cranial ectoderm, implicating FGF8 as a central component in generic placode induction. PMID- 16271715 TI - The K-Ras 4A isoform promotes apoptosis but does not affect either lifespan or spontaneous tumor incidence in aging mice. AB - Ras proteins function as molecular switches in signal transduction pathways, and, here, we examined the effects of the K-ras4A and 4B splice variants on cell function by comparing wild-type embryonic stem (ES) cells with K ras(tmDelta4A/tmDelta4A) (exon 4A knock-out) ES cells which express K-ras4B only and K-ras(-/-) (exons 1-3 knock-out) ES cells which express neither splice variant, and intestinal epithelium from wild-type and K-ras(tmDelta4A/tmDelta4A) mice. RT-qPCR analysis found that K-ras4B expression was reduced in K ras(tmDelta4A/tmDelta4A) ES cells but unaffected in small intestine. K-Ras deficiency did not affect ES cell growth, and K-Ras4A deficiency did not affect intestinal epithelial proliferation. K-ras(tmDelta4A/tmDelta4A) and K-ras(-/-) ES cells showed a reduced capacity for differentiation following LIF withdrawal, and K-ras(-/-) cells were least differentiated. K-Ras4A deficiency inhibited etoposide-induced apoptosis in ES cells and intestinal epithelial cells. However, K-ras(tmDelta4A/tmDelta4A) ES cells were more resistant to etoposide-induced apoptosis than K-ras(-/-) cells. The results indicate that (1) K-Ras4A promotes apoptosis while K-Ras4B inhibits it, and (2) K-Ras4B, and possibly K-Ras4A, promotes differentiation. The findings raise the possibility that alteration of the K-Ras4A/4B isoform ratio modulates tumorigenesis by differentially affecting stem cell survival and/or differentiation. However, K-Ras4A deficiency did not affect life expectancy or spontaneous overall tumor incidence in aging mice. PMID- 16271716 TI - Persistent motor deficit following infusion of autologous blood into the periventricular region of neonatal rats. AB - Periventricular hemorrhage (PVH) in the brain of premature infants is often associated with developmental delay and persistent motor deficits. Our goal is to develop a rodent model that mimics the behavioral phenotype. We hypothesized that autologous blood infusion into the periventricular germinal matrix region of neonatal rats would lead to immediate and long-term behavioral changes. Tail blood or saline was infused into the periventricular region of 1-day-old rats. Magnetic resonance (MR) imaging was used to demonstrate the hematoma. Rats with blood infusion, as well as saline and intact controls, underwent behavior tests until 10 weeks age. Blood-infused rats displayed significant delay in motor development (ambulation, righting response, and negative geotaxis) to 22 days of age. As young adults, they exhibited impaired ability to stay on a rotating rod and to reach for food pellets. MR imaging at 10 weeks demonstrated subsets of rats with normal appearing brains, focal cortical infarcts, or mild hydrocephalus. There was a good correlation between MR imaging and histological findings. Some rats exhibited periventricular heterotopia and/or subtle striatal abnormalities not apparent on MR images. We conclude that autologous blood infusion into the brain of neonatal rats successfully models some aspects of periventricular hemorrhage that occurs after premature birth in humans. PMID- 16271717 TI - Trypanosoma rangeli: effects of physalin B on the immune reactions of the infected larvae of Rhodnius prolixus. AB - Physalins are seco-steroids obtained from plants of the family Solanaceae. Herein, we tested Physalis angulata L purified physalin B as an immunomodulatory compound in 5th-instar larvae of Rhodnius prolixus, which were systemically infected with the H14 Trypanosoma rangeli strain protozoan. In uninfected insects, the effective concentration of physalin B, which inhibited 50% of the blood ingested (ED(50)) volume, was 15.2+/-1.6 microg/ml of the meal. Ecdysis processes and mortality in uninfected larvae, treated orally with physalin B in concentrations ranging from 1 to 10 microg/ml, was similar to that observed in insects not treated with physalin B. However, R. prolixus larvae previously fed on blood containing 1.0, 0.1, and 0.01 microg of physalin B/ml exhibited mortality rates of 78.1, 54.3, and 12.7%, respectively, 6 days after inoculation of T. rangeli (1 x 10(3) parasites/insect), whereas only 7.2% mortality was observed in the control group, injected with sterile culture medium. The insects treated with physalin B (0.1 microg/ml) and inoculated with T. rangeli did not modify the phenoloxidase (PO) activity and total hemocyte count in the hemolymph. However, physalin B treatment caused a reduction in hemocyte micro-aggregation and nitric oxide production and enhanced the parasitemia in the hemolymph. These results demonstrate that physalin B from P. angulata is a potent immunomodulatory substance for the bloodsucking insect, R. prolixus. PMID- 16271718 TI - A tRNA(Glu) that uncouples protein and tetrapyrrole biosynthesis. AB - Glu-tRNA is either bound to elongation factor Tu to enter protein synthesis or is reduced by glutamyl-tRNA reductase (GluTR) in the first step of tetrapyrrole biosynthesis in most bacteria, archaea and in chloroplasts. Acidithiobacillus ferrooxidans, a bacterium that synthesizes a vast amount of heme, contains three genes encoding tRNA(Glu). All tRNA(Glu) species are substrates in vitro of GluRS1 from A. ferrooxidans.Glu-tRNA(3)(Glu), that fulfills the requirements for protein synthesis, is not substrate of GluTR. Therefore, aminoacylation of tRNA(3)(Glu) might contribute to ensure protein synthesis upon high heme demand by an uncoupling of protein and heme biosynthesis. PMID- 16271719 TI - Antibiotic susceptibility of mammalian mitochondrial translation. AB - All medically useful antibiotics should have the potential to distinguish between target microbes (bacteria) and host cells. Although many antibiotics that target bacterial protein synthesis show little effect on the translation machinery of the eukaryotic cytoplasm, it is unclear whether these antibiotics target or not the mitochondrial translation machinery. We employed an in vitro translation system from bovine mitochondria, which consists of mitochondrial ribosomes and mitochondrial elongation factors, to estimate the effect of antibiotics on mitichondrial protein synthesis. Tetracycline and thiostrepton showed similar inhibitory effects on both Escherichia coli and mitochondrial protein synthesis. The mitochondrial system was more resistant to tiamulin, macrolides, virginiamycin, fusidic acid and kirromycin than the E. coli system. The present results, taken together with atomic structure of the ribosome, may provide useful information for the rational design of new antibiotics having less adverse effects in humans and animals. PMID- 16271720 TI - Antepartum pelvic support defects in nulliparous and multiparous women. PMID- 16271722 TI - Critical nucleus size effects on nanoparticle formation in microemulsions: a comparison study between experimental and simulation results. AB - Monte Carlo simulations were performed to study the influence of critical nucleus size on nanoparticle formation in microemulsions. It was found that critical nucleus size strongly affected nucleation and growth rates, as well as final nanoparticle sizes. An increase of critical nucleus leads to a slower nucleation process. In contrast, it gives rise to acceleration of the growth process. Final nanoparticle sizes increase as the critical nucleus value increases. It is predicted that this dependence will be less pronounced when a high reactant concentration is used. We have compared the simulation results with experimental data taken from different authors. Good agreement between the two kinds of results supports the conclusions of this paper. PMID- 16271721 TI - Oral misoprostol in the third stage of labor. AB - OBJECTIVE: To compare the efficacy of intravenous ergometrine, intramuscular oxytocin, and oral misoprostol in the control of postpartum hemorrhage. METHODS: Mean blood loss, rates of blood loss between 500 and 1000 ml, hematocrit fall greater than 10%, and need for additional oxytocic agents and nature and rates of adverse effects were assessed in this prospective, randomized, controlled study. RESULTS: All outcomes were similar in the 3 groups. The main adverse effects in the misoprostol group were temperatures higher than 99 degrees F, which normalized within 2 h and shivering, which was mild and self-limiting. CONCLUSIONS: Oral misoprostol is as effective as conventional oxytocic agents in preventing postpartum hemorrhage and can be recommended for use in low-resource settings. PMID- 16271723 TI - A strategy of retrograde injection of bone marrow mononuclear cells into the myocardium for the treatment of ischemic heart disease. AB - OBJECTIVE: Bone marrow cells implantation (BMI) has been reported to efficiently improve ischemic heart disease. However, BMI strategies are generally invasive. To establish a BMI strategy for ischemic heart disease, we performed implantation of autologous cryopreserved mononuclear cells (MNCs) from bone marrow (BM) retrogradely into the myocardium via the coronary vein in pigs with acute myocardial infarction (AMI) and old myocardial infarction (OMI). METHODS: BM cells were harvested from the pigs' fumurs. MNCs were collected by centrifugation and were cryopreserved. Anterior myocardial infarction was induced by occlusion of the midportion of the left anterior descending coronary artery without surgical intervention. Frozen BM cells were quickly thawed and injected retrogradely via the coronary vein into the myocardium through a single balloon infusion catheter 6 h and 2 weeks after the induction of infarction. Four weeks after implantation, coronary arteriograms were obtained, cardiac function was analyzed with the use of a conductance catheter, and histopathologic analysis was performed with a confocal laser microscope. Plasma levels of natriuretic peptides and angiogenic growth factors were measured after BMI. RESULTS: Flow cytometric analysis revealed that 90% of cryopreserved BM cells were viable in vitro. Labeled BM cells were entirely distributed around in the infarcted area of maycardium in pigs. BMI increased collateral neovascuralization in infarcted hearts. BMI significantly improved cardiac function in AMI with BMI and OMI with BMI groups. BMI also increased the formation of microcapillary arteries in infarcted hearts. Levels of natriuretic peptides were significantly decreased, and levels of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (FGF2) were significantly increased after BMI. Confocal laser microscopy revealed the presence of proliferative and activated myocardial cells in infarcted hearts after BMI. CONCLUSION: The retrograde infusion of cryopreserved BM cells into myocardium efficiently induced angiogenesis and improved cardiac function in pigs with AMI or OMI. These results suggest that the present strategy of BMI will be safe and feasible as an angiogenic cell therapy for ischemic heart disease. PMID- 16271724 TI - An intronic peroxisome proliferator-activated receptor-binding sequence mediates fatty acid induction of the human carnitine palmitoyltransferase 1A. AB - The liver plays a central role in the response to fasting. The hormonal profile in this condition, low insulin, and high concentrations of glucagon in plasma, induce the release of large amounts of fatty acids from adipose tissue. Prolonged starvation can therefore induce a dramatic change in the fatty acid oxidative capacity of liver metabolism. Modulation of gene expression by PPARalpha plays a crucial role in this response. While a major role for PPARalpha in the liver is to produce ketone bodies as fuel through beta-oxidation for peripheral tissues during fast, its participation in the control of CPT1A, the rate-limiting step of the pathway, remains controversial. Using Web-based software (VISTA) combining transcription factor binding site database searches with comparative sequence analyses, we have localized a conserved functional PPAR responsive element downstream of the transcriptional start site of the human CPT1A gene. We have shown that this sequence is fundamental for fatty acids or PGC1-induced transcriptional activation of the CPT1A gene. These results corroborate the hypothesis that PPARalpha regulates the limiting step in the oxidation of fatty acids in liver mitochondria. PMID- 16271725 TI - Structure and orientation of peptide inhibitors bound to beta-amyloid fibrils. AB - Polymerization of the soluble beta-amyloid peptide into highly ordered fibrils is hypothesized to be a causative event in the development of Alzheimer's disease. Understanding the interactions of Abeta with inhibitors on an atomic level is fundamental for the development of diagnostics and therapeutic approaches, and can provide, in addition, important indirect information of the amyloid fibril structure. We have shown recently that trRDCs can be measured in solution state NMR for peptide ligands binding weakly to amyloid fibrils. We present here the structures for two inhibitor peptides, LPFFD and DPFFL, and their structural models bound to fibrillar Abeta(14-23) and Abeta(1-40) based on transferred nuclear Overhauser effect (trNOE) and transferred residual dipolar coupling (trRDC) data. In a first step, the inhibitor peptide structure is calculated on the basis of trNOE data; the trRDC data are then validated on the basis of the trNOE-derived structure using the program PALES. The orientation of the peptide inhibitors with respect to Abeta fibrils is obtained from trRDC data, assuming that Abeta fibrils orient such that the fibril axis is aligned in parallel with the magnetic field. The trRDC-derived alignment tensor of the peptide ligand is then used as a restraint for molecular dynamics docking studies. We find that the structure with the lowest rmsd value is in agreement with a model in which the inhibitor peptide binds to the long side of an amyloid fibril. Especially, we detect interactions involving the hydrophobic core, residues K16 and E22/D23 of the Abeta sequence. Structural differences are observed for binding of the inhibitor peptide to Abeta14-23 and Abeta1-40 fibrils, respectively, indicating different fibril structure. We expect this approach to be useful in the rational design of amyloid ligands with improved binding characteristics. PMID- 16271727 TI - Impacts of experimental dredged material disposal on a shallow, sublittoral macrofauna community in Mecklenburg Bay (western Baltic Sea). AB - An experimental disposal of dredged material was performed in June 2001 at two adjacent sites in Mecklenburg Bay (western Baltic Sea) to investigate the impacts on a sublittoral fauna community (macrozoobenthos). Temporal changes in community parameters were analysed: Total abundance and multidimensional scaling (MDS) ordination results showed that dredged material disposal causes significant changes in benthic assemblages in comparison to the original situation before the disposal activities. The bivalves Arctica islandica and Macoma balthica were almost not affected by the disposal. Abundances of the polychaete N. hombergii as well as the epibenthic cumacean D. rathkei decreased by a factor of 1.4-3.9 whereas most other invertebrates suffered even more. Two years after the experimental disposal a high level of benthic recovery was found, but also a change in dominance structure. These changes are rather due to inter-annual variability in regional benthic recruitment processes in the Mecklenburg Bay than due to the disposal. PMID- 16271726 TI - Duloxetine for the treatment of major depressive disorder: a closer look at efficacy and safety data across the approved dose range. AB - OBJECTIVE: This analysis focuses on efficacy and safety data obtained from studies of duloxetine for the treatment of major depressive disorder (MDD) within the approved dose range of 40-60 mg/day. METHOD: Efficacy and safety data were obtained from the acute phase portions of four randomized, double-blind, placebo controlled clinical trials in patients meeting DSM-IV criteria for MDD. In Studies 1 and 2, patients were randomized to duloxetine 60 mg once daily (QD) (n=123 [Study 1]; n=128 [Study 2]) or placebo (n=122 [Study 1]; n=139 [Study 2]) for 9 weeks. In Studies 3 and 4, patients were randomized to duloxetine 20 mg twice daily (BID) (n=91 [Study 3]; n=86 [Study 4]) or placebo (n=90 [Study 3]; n=89 [Study 4]) for 8 weeks. Efficacy measures included the 17-item Hamilton Rating Scale for Depression (HAMD17) total score (primary outcome), HAMD17 subscales, the Clinical Global Impression of Severity (CGI-S) and Patient Global Impression of Improvement (PGI-I) scales, and Visual Analog Scales (VAS) for pain. Safety assessments included rates of discontinuation due to adverse events, spontaneously reported treatment-emergent adverse events, and changes in vital signs. RESULTS: In both studies of duloxetine 60 mg QD, mean change in HAMD17 total score was significantly greater in duloxetine-treated patients compared with placebo (Study 1, p<.001; Study 2, p=.024). At a dose of 20 mg BID, duloxetine demonstrated significant superiority over placebo on the HAMD17 total score in one of the two studies (Study 4, p=.034). Probabilities of remission among patients receiving duloxetine 60 mg QD were 44.2% in Study 1 (p<.001 vs. placebo) and 43.0% in Study 2 (NS), while for patients receiving duloxetine 20 mg BID the probabilities of remission were 27.2% in Study 3 (NS) and 36.1% in Study 4 (NS). Across the six assessed VAS measures of pain severity and interference, the main effect of treatment for duloxetine 60 mg QD was significantly superior to placebo on 7 of the 12 outcomes in Studies 1 and 2, while duloxetine 20 mg BID was not superior to placebo on any of the 12 outcomes in Studies 3 and 4. The rate of discontinuation due to adverse events was 13.1% among patients receiving duloxetine 60 mg QD, and 11.9% at a dose of 20 mg BID. The most frequently reported treatment-emergent adverse events at both doses included nausea, headache, dry mouth, dizziness, and insomnia. The incidence of treatment-emergent nausea among patients receiving duloxetine 60 mg QD was 37.8%, compared with 16.4% among patients receiving 20 mg BID. CONCLUSION: Duloxetine provides safe and effective acute phase treatment of MDD at doses of 40-60 mg/day. Compared with placebo, the 60 mg QD dose was more consistently effective than the 20 mg BID dose. However, the incidence of certain treatment-emergent adverse events is likely to be lower at the 40 mg dose. PMID- 16271729 TI - A benthic survey of Aliwal Shoal and assessment of the effects of a wood pulp effluent on the reef. AB - Aliwal Shoal lies south of Durban in South Africa and has been the subject of recent bathymetric, seafloor and benthic surveys. ANOVA of the biological data revealed that the biota were uniformly distributed on the reef with the exception of encrusting sponges and algae on rock. The variations in distribution of these biota were significant and, in the case of the encrusting sponges, appeared to be related to the discharge of a wood pulp effluent. Further evidence of this was suggested by stable isotope analyses of representative organisms. The encrusting sponges were recommended as good candidates for further monitoring of the effects of the wood pulp effluent on Aliwal Shoal as the effluent pipeline has been extended. PMID- 16271728 TI - Characterization of the ecological quality of the coastal Gulf of Lions (NW Mediterranean). A comparative approach based on three biotic indices. AB - The ecological quality of the Gulf of Lions coast was assessed using three biotic indices (H', AMBI and BQI). The three indices correlated positively. The positive correlation between AMBI and BQI was surprising and was mostly due to the fact that the dominant species Ditrupa arietina featured a low ES50(0.05) but was classified in GI by AMBI. Both H' and BQI were efficient in distinguishing impacted from un-impacted sites but AMBI was not. Differences between H' and BQI were mainly due to the scale used to translate indices in terms of EcoQ. The three indices were able to detect the major changes in macrofauna composition, which occurred in the Bay of Banyuls-sur-Mer during the last 40years. However, the interpretations of such changes in terms of EcoQ differed between indices. These results are discussed relative to the characteristics of the tested indices. PMID- 16271730 TI - Sources and transport of hydrocarbons in sediments from the Changjiang River Estuary, China. PMID- 16271731 TI - The ECOMAN project: A novel approach to defining sustainable ecosystem function. AB - The ECOMAN was initiated in 2001 by the University of Plymouth, UK, Plymouth Marine Laboratory and the Department of the Environment, Fisheries and Rural Affairs (DEFRA) to address the need for more pragmatic assessment techniques linking environmental degradation with its causes. The primary aim of the project was to develop an evidence-based approach in which suites of easy-to-use, cost effective and environmentally valid biological responses (biomarkers) could be used together to assess the health of coastal systems through the general condition of individuals. A range of sub-lethal endpoints, chosen to reflect successive levels of biological organisation (molecular, cellular, physiological), was evaluated in common coastal organisms showing different feeding types (filter feeding, grazing, predation) and habitat requirements (estuary, rocky shore). Initially, the suite of biomarkers was used in laboratory studies to determine the relative sensitivities of key species within different functional groups to common contaminants. These results were then validated in field studies performed in a range of ecosystems exhibiting different degrees/signatures of contamination. Here, an example is provided of a field study in the Humber Estuary, UK, which illustrates how multivariate statistical analysis can be used to identify patterns of response to discriminate between contaminated and clean sites. The use of a holistic, integrated approach of this kind is advocated as a practical means of assessing the impact of chemical contamination on organismal health and of ranking the status of marine ecosystems. PMID- 16271732 TI - Heavy metal contamination in mangrove habitats of Singapore. PMID- 16271733 TI - Algicidal and antifungal compounds from the roots of Ruta graveolens and synthesis of their analogs. AB - Bioassay-guided fractionation of the ethyl acetate extract of Ruta graveolens roots yielded rutacridone epoxide with potent selective algicidal activity towards the 2-methyl-isoborneol (MIB)-producing blue-green alga Oscillatoria perornata, with relatively little effect on the green alga Selenastrum capricornutum. The diol-analog of rutacridone epoxide, gravacridondiol, which was also present in the same extract, had significantly less activity towards O. perornata. Rutacridone epoxide also showed significantly higher activity than commercial fungicides captan and benomyl in our micro-bioassay against the agriculturally important pathogenic fungi Colletotrichum fragariae, C. gloeosporioides, C. acutatum, and Botrytis cineara and Fusarium oxysporium. Rutacridone epoxide is reported as a direct-acting mutagen, precluding its use as an agrochemical. In order to understand the structure-activity relationships and to develop new potential biocides without toxicity and mutagenicity, some analogs containing the (2-methyloxiranyl)-dihydrobenzofuran moiety with an epoxide were synthesized and tested. None of the synthetic analogs showed comparable activities to rutacridone epoxide. The absolute stereochemistry of rutacridone was determined to be 2'(R) and that of rutacridone epoxide to be 2'(R), 3'(R) by CD and NMR analysis. PMID- 16271734 TI - Phenolic glycosides and ionone glycoside from the stem of Sargentodoxa cuneata. AB - Four phenolic glycosides, cuneatasides A-D (1-4), and one ionone glycoside cuneataside E (5), together with seven known phenolic compounds (6-12) were isolated from the water-soluble constituents of the stem of Sargentodoxa cuneata (Sargentodoxaceae). Their structures were elucidated by spectroscopic analysis. In vitro tests for antimicrobial activity showed compounds 1 and 2 to possess significant activity against two Gram-positive organisms, Staphylococcus aureus and Micrococcus epidermidis. PMID- 16271735 TI - Ghrelin-stimulation test in the diagnosis of canine pituitary dwarfism. AB - This study investigated whether ghrelin, a potent releaser of growth hormone (GH) secretion, is a valuable tool in the diagnosis of canine pituitary dwarfism. The effect of intravenous administration of ghrelin on the release of GH and other adenohypophyseal hormones was investigated in German shepherd dogs with congenital combined pituitary hormone deficiency and in healthy Beagles. Analysis of the maximal increment (i.e. difference between pre- and maximal post-ghrelin plasma hormone concentration) indicated that the GH response was significantly lower in the dwarf dogs compared with the healthy dogs. In none of the pituitary dwarfs, the ghrelin-induced plasma GH concentration exceeded 5 microg/l at any time. However, this was also true for 3 healthy dogs. In all dogs, ghrelin administration did not affect the plasma concentrations of ACTH, cortisol, TSH, LH and PRL . Thus, while a ghrelin-induced plasma GH concentration above 5 microg/l excludes GH deficiency, false-negative results may occur. PMID- 16271736 TI - The ideal free distribution and bacterial growth on two substrates. AB - A population dynamical model describing growth of bacteria on two substrates is analyzed. The model assumes that bacteria choose substrates in order to maximize their per capita population growth rate. For batch bacterial growth, the model predicts that as the concentration of the preferred substrate decreases there will be a time at which both substrates provide bacteria with the same fitness and both substrates will be used simultaneously thereafter. Preferences for either substrate are computed as a function of substrate concentrations. The predicted time of switching is calculated for some experimental data given in the literature and it is shown that the fit between predicted and observed values is good. For bacterial growth in the chemostat, the model predicts that at low dilution rates bacteria should feed on both substrates while at higher dilution rates bacteria should feed on the preferred substrate only. Adaptive use of substrates permits bacteria to survive in the chemostat at higher dilution rates when compared with non-adaptive bacteria. PMID- 16271737 TI - Evolutionarily stable strategies and short-term selection in Mendelian populations re-visited. AB - This note concerns a one locus, two allele, random mating diploid population, subject to frequency-dependent viability selection. It is already known that in such a population, any evolutionarily stable strategies (ESS), if only accessible by the genotype-to-phenotype mapping, is the phenotypic image of a stable genetic equilibrium (Eshel, I. 1982. Evolutionarily stable strategies and viability selection in Mendelian populations. Theor. Popul. Biol. 22(2), 204-217; Cressman et al. 1996. Evolutionary stability in strategic models of single-locus frequency dependent viability selection. J. Math. Biol. 34, 707-733). The opposite is not true. We find necessary and sufficient parametric conditions for global convergence to the ESS, but we also demonstrate conditions under which, although a unique, genetically accessible ESS exists, there is another, "non-phenotypic" genetically stable equilibrium. PMID- 16271738 TI - Inhibition of tumor necrosis factor-alpha-induced expression of adhesion molecules in human endothelial cells by the saponins derived from roots of Platycodon grandiflorum. AB - Adhesion molecules play an important role in the development of atherogenesis and are produced by endothelial cells after being stimulated with various inflammatory cytokines. This study examined the effect of saponins that were isolated from the roots of Platycodon grandiflorum A. DC (Campanulaceae), Changkil saponins (CKS), on the cytokine-induced monocyte/human endothelial cell interaction, which is a crucial early event in atherogenesis. CKS significantly inhibited the TNFalpha-induced increase in monocyte adhesion to endothelial cells as well as decreased the protein and mRNA expression levels of vascular adhesion molecule-1 and intercellular cell adhesion molecule-1 on endothelial cells. Furthermore, CKS significantly inhibited the TNFalpha-induced production of intracellular reactive oxygen species (ROS) and activation of NF-kappaB by preventing IkappaB degradation and inhibiting IkappaB kinase activity. Overall, CKS has anti-atherosclerotic and anti-inflammatory activity, which is least in part the result of it reducing the cytokine-induced endothelial adhesion to monocytes by inhibiting intracellular ROS production, NF-kappaB activation, and cell adhesion molecule expression in endothelial cells. PMID- 16271739 TI - Redefining the common insertion site. AB - Retroviral mutagenesis has been used as a powerful tool to discover genes involved in oncogenesis through a technique called Common Insertion Site (CIS) analysis where tumors are induced by proviral integrations and the genomic loci of the proviruses are identified. A fundamental assumption made in this analysis is that multiple proviral insertions in close proximity occurring more frequently than would be predicted randomly provides evidence that the genes near the integrations are involved in the formation of the tumors. We demonstrate here using data derived from MLV integrations not put under selection for tumor induction that CIS analysis as currently defined is often not a sufficient argument for a gene's significance in tumorigenesis. PMID- 16271740 TI - Ceramide regulates SR protein phosphorylation during adenoviral infection. AB - In this study, we show that adenoviral infection induced accumulation of the sphingolipid ceramide in a dose- and time-dependent manner. This accumulation preceded cell lysis, occurred in the absence of biochemical evidence of apoptosis, and was derived from de novo synthesis of ceramide. An adenovirus mutant that lacks the adenovirus death protein (ADP) produced ceramide accumulation in the absence of cell lysis. This suggested that ceramide accumulation was either driven by adenovirus or was a cellular stress response but was unlikely a result of cell death. The use of inhibitors of ceramide synthesis resulted in a significant delay in cell lysis, suggesting that ceramide was necessary for the lytic phase of the infection. Serine/arginine-rich (SR) proteins were dephosphorylated during the late phase of the viral cycle, and inhibitors of ceramide synthesis reversed this. These findings suggest that adenovirus utilizes the ceramide pathway to regulate SR proteins during infection. PMID- 16271741 TI - Selective elimination of HIV-1-infected cells by Env-directed, HIV-1-based virus like particles. AB - We recently showed that both replicating and resting cells cultivated with ganciclovir (GCV) were killed when challenged with vesicular stomatitis virus G glycoprotein pseudotyped HIV-1-based virus-like particles (VLPs) carrying the Nef7 (i.e., an HIV-1 Nef mutant incorporating in virions at high levels)/herpes simplex virus-1 thymidine kinase (HSV-TK) fusion product. On this basis, a novel anti-HIV therapeutic approach based on Nef7/TK VLPs expressing X4 or R5 HIV cell receptor complexes has been attempted. We here report that (CD4-CXCR4) and (CD4 CCR5) Nef7-based VLPs efficiently enter cells infected by X4- or R5-tropic HIV-1 strains, respectively. Importantly, the delivery of the VLP-associated Nef7/TK led to cell death upon GCV treatment. Of interest, VLPs were effective also against non-replicating, HIV-1-infected primary human monocyte-derived macrophages. HIV-targeted VLPs represent a promising candidate for the treatment of persistently HIV-1-infected cells that are part of virus reservoirs resistant to HAART therapies. PMID- 16271742 TI - Determining whether a ball will land behind or in front of you: not just a combination of expansion and angular velocity. AB - We propose and evaluate a source of information that ball catchers may use to determine whether a ball will land behind or in front of them. It combines estimates for the ball's horizontal and vertical speed. These estimates are based, respectively, on the rate of angular expansion and vertical velocity. Our variable could account for ball catchers' data of Oudejans et al. [The effects of baseball experience on movement initiation in catching fly balls. Journal of Sports Sciences, 15, 587-595], but those data could also be explained by the use of angular expansion alone. We therefore conducted additional experiments in which we asked subjects where simulated balls would land under conditions in which both angular expansion and vertical velocity must be combined for obtaining a correct response. Subjects made systematic errors. We found evidence for the use of angular velocity but hardly any indication for the use of angular expansion. Thus, if catchers use a strategy that involves combining vertical and horizontal estimates of the ball's speed, they do not obtain their estimates of the horizontal component from the rate of expansion alone. PMID- 16271743 TI - Pictorial cues constrain depth in da Vinci stereopsis. AB - "da Vinci stereopsis" is defined as depth seen in a monocular object occluded by a binocular one, and the visual system must solve its depth ambiguity [Nakayama, K., & Shimojo, S. (1990). da Vinci stereopsis: Depth and subjective occluding contours from unpaired image points. Vision Research, 30, 1811-1825]. Although fused images include various pictorial features, effects of pictorial depth cues have never been systematically investigated in da Vinci stereopsis. To examine this, we created stereograms consisting of a monocular bar flanked by binocular bars with a fixed large horizontal separation, in which the monocular bar induced a subjective occluding edge. Manipulating vertical size or contrast of the bars could affect the depth of the monocular bar. Conflicting these cues revealed that the effect of vertical size was stronger than that of contrast in all our subjects. Measurements of the depth indicated that the relative vertical size of the bars quantitatively determined the perceived depth, of which levels had large inter-subject differences. All these experiments indicate that the visual system can use the pictorial depth cues as a constraint to determine the depth of monocular elements. PMID- 16271744 TI - Eye movements during multiple readings of the same text. AB - People often read the same text more than once. Studying eye movements during multiple readings of the same texts provides a unique opportunity to observe the consistency of saccadic landing positions. Eye movements were recorded while 5 people read the same 4 texts more than 40 times, no more than 4 times/day, and never on consecutive trials. Other texts, read only once, were interspersed. Comprehension questions and a change-detection task helped maintain attention in the face of the repetition. There were two main findings: (1) repeated reading produced significant, but modest, changes in global saccadic patterns. The only change found in all readers was a reduction in the proportion of regressions. (2) Saccadic landing positions fell into clusters located at a variety of places with respect to word boundaries, and often across word boundaries. A mixed-strategy model of saccadic guidance (look to the center of words, while trying to maintain fairly uniform saccade lengths), could account for the overall strength of clustering, but not for the variability among cluster locations, suggesting that saccadic landing sites are selected in part on the basis of local text characteristics. The reliable clustering of saccadic landing positions found during multiple readings of the same text opens the way for cluster patterns to be used to study eye movement strategies during reading and overcome at least some of the variability associated with traditional global single-text measures. PMID- 16271745 TI - Anatomical descriptions, comparative studies and evolutionary significance of the hominin skulls from Dmanisi, Republic of Georgia. AB - Evidence for ancient hominin occupation in Eurasia comes from Dmanisi in the Georgian Caucasus. Stratigraphic and sedimentological arguments, geochemical observations, paleomagnetic sampling and radiometric dates all point to the conclusion that bones and artifacts were deposited at this site during a brief interval following the close of the Olduvai Subchron (1.77 million years ago). In this report we present further descriptive and comparative studies of the D2280 braincase, the D2282 partial cranium, now linked with the D211 mandible, and the skull D2700/D2735. The crania have capacities ranging from 600 cm3 to 775 cm3. Supraorbital tori and other vault superstructures are only moderately developed. The braincase is expanded laterally in the mastoid region, but the occiput is rounded. The pattern of sagittal keeling is distinctive. D2700 displays a prominent midfacial profile and has a very short nasoalveolar clivus. Also, the M3 crowns are reduced in size. Although there is variation probably related to growth status and sex dimorphism, it is appropriate to group the Dmanisi hominins together. With the possible exception of the large D2600 mandible, the individuals are sampled from one paleodeme. This population resembles Homo habilis in brain volume and some aspects of craniofacial morphology, but many of these features can be interpreted as symplesiomorphies. Other discrete characters and measurements suggest that the Dmanisi skulls are best placed with H. erectus. There are numerous similarities to individuals from the Turkana Basin in Kenya, but a few features link Dmanisi to Sangiran in Java. Some traits expressed in the Dmanisi assemblage appear to be unique. Reconstructing the evolutionary relationships of these ancient populations of Africa and Eurasia is difficult, as the record is quite patchy, and determination of character polarities is not straightforward. Nevertheless, the evidence from anatomical analysis and measurements supports the hypothesis that Dmanisi is close to the stem from which H. erectus evolved. PMID- 16271746 TI - Trends and interactions of physical and bio-geo-chemical features in the Adriatic Sea as derived from satellite observations. AB - Time series of satellite data, generated by the AVHRR (1981-1999), CZCS (1979 1985) and SeaWiFS (1998-2002), have been used to assess trends and interactions of physical and bio-geo-chemical features in the Adriatic Sea. The images were processed to estimate Sea Surface Temperature (SST) and Chlorophyll-like Pigment Concentration (CPC). Long-term composites and climatologies were derived, using fixed geographical grids and projections. The AVHRR data show an apparent warming trend, when plotting the sequence of seasonal cycles (monthly mean SST, averaged over the whole basin) against time, due to a steady rise of summer values. Considering 3 regions (north, central and south), split into east and west sections, the northern Adriatic shows high SST fluctuations (possibly associated with the cycle of winter cooling and summer warming, typical of the relatively shallow sub-basin), while the southern Adriatic exhibits a lower variability (possibly influenced by the periodic water incoming from, and outflowing to the Ionian Sea). During summer, an east-west gradient prevails, while during winter only a general north-south gradient can be found. The SeaWiFS-derived CPC values, distributions and trends appear to be consistent with the historical CZCS record. Persistent differences in the quantitative assessment of CPC for coastal waters is due to the use of improved algorithms, less influenced by the presence of dissolved organics and suspended sediments in the water column, for the processing of SeaWiFS data. Apparent incongruities of the space and time patterns in the SeaWiFS record with respect to the reference climatology, obtained by CZCS more than a decade before, occur chiefly when considering the spring bloom in the southern Adriatic and the summer development of the north Adriatic front. The comparison of the long-term times series of satellite data shows that there is a high correlation between patterns in the thermal field and in the colour field. This suggests that different surface waters, identified by the SST index, are also traced by different ecological features, identified by the CPC index. Both indices also show a high correlation with the classical cyclonic circulation scheme of the Adriatic Sea, proposing once again an intimate relationship between the water dynamics and its bio-geo-chemistry. PMID- 16271747 TI - Mercury distribution in the skin of beluga (Delphinapterus leucas) and narwhal (Monodon monoceros) from the Canadian Arctic and mercury burdens and excretion by moulting. AB - Beluga and narwhal skin as a whole (in Inuktitut known as "muktuk") is considered to be a delicacy by native Canadian and Greenland people. Individual strata of the skin, and muscle from 27 beluga from the western, and 20 narwhal from the eastern Canadian Arctic, were analyzed for mercury and the thickness and density of each skin layer was measured. Mercury was not uniformly distributed in the skin, but increased outwardly with each layer. The concentration was only 0.29 and 0.16 microg/g (wet wt) in the innermost layer (dermis) of belugas and narwhal respectively, and 1.5 and 1.4 microg/g (wet wt) in the outermost layer (degenerative epidermis) of beluga and narwhal, respectively. There was a significant (alpha=0.05) association between age and mercury concentration in each skin layer, the regression coefficients progressively increasing from the inner layer (dermis) to the outer layer: 0.011-0.063 microg/g year-1; 0.034 microg/g year-1 for skin as a whole; 0.054 microg/g year-1 for muscle. The concentration of total mercury was 0.84 and 0.59 microg/g (wet wt) in skin as a whole (muktuk) of beluga and narwhal respectively, and 0.12 and 0.03 microg/g in blubber, respectively. The average, total mercury concentration in muscle tissue was 1.4 and 0.81 microg/g wet wt, in beluga and narwhal respectively, exceeding (except for blubber) the Canadian Government's Guideline (0.5 microg/g wet wt) for fish export and consumption. The skin surface area of an average-size beluga and narwhal was estimated (6.10 and 6.50 m2, respectively), as were excretions of mercury through moulting (13,861 and 6721 microg year-1; 14 and 7 mg year-1) for belugas and narwhal, respectively. The whole-body mercury burden (699,300 microg; 700 mg) for a 1000 kg beluga and its various tissues were estimated, as was the fraction of mercury excreted by moulting (2-0.42% of the whole-body burden). Annual mercury burden increments in beluga skin, muscle and the whole body were estimated (2750; 17,280; 40,00 microg year-1, respectively), using regression coefficients of age on mercury concentration. The annual gross mercury intake via food was estimated (131,400 microg), of which 70% was excreted. PMID- 16271748 TI - Clinical features and outcomes of uterine and ovarian carcinosarcoma. AB - BACKGROUND: The objective of this study was to compare the clinical presentation and outcomes of women with ovarian and uterine carcinosarcoma (CS). METHODS: We performed a retrospective review of patients treated for uterine or ovarian CS from 1952 to 2003. Fisher's Exact Test was used to compare patient characteristics. Survival curves were estimated using the Kaplan-Meier method and compared using the log rank test. RESULTS: We identified 87 patients with uterine CS and 18 with ovarian CS. There was no difference in age, body mass index, parity, menopausal status, family history of cancer, history of pelvic radiation, diabetes or hypertension between the two groups. 43% of women with uterine CS presented at stage I/II, compared to 28% of women with ovarian tumors (P = 0.0003). 82% of patients with ovarian tumors received adjuvant chemotherapy with or without radiation; 51% of the patients in the uterine CS group received adjuvant radiation therapy. The median length of follow-up was 13 months. There was no difference in the Kaplan-Meier estimates of overall survival between the two disease sites. The median survival for uterine CS patients was 16 months, compared to 11 months in the ovarian CS group; HR = 0.991 (95% CI = 0.534, 1.839). CONCLUSIONS: We found no differences in patient demographics between the two groups. Despite differences in stage and initial treatment, there was no difference in survival between women with uterine and ovarian CS. PMID- 16271749 TI - TP53 overexpression in recurrent endometrial carcinoma. AB - OBJECTIVE: To study alterations within the p53 pathway in relation to the development of recurrent stage I endometrioid endometrial carcinoma. METHODS: Paraffin-embedded tumor tissue of both primary and recurrent tumors from 44 patients with and 44 without recurrence was used for immunohistochemical analysis of TP53, hMdm2, P21(Waf1/Cip1) and M30. DNA was extracted, and mutation analysis of p53 (exon 5-8, 11) was performed by direct sequencing. RESULTS: TP53 overexpression was significantly associated with recurrent disease: Odds Ratio 3.8 (95% CI: 1.5-9.8). Overexpression of TP53 was associated with lower staining indices (SI:0-9) of both hMdm2 and P21 in tumors of patients with recurrence, compared to controls: 2.0 +/- 0.4 vs. 4.0 +/- 0.8 and 1.9 +/- 0.8 vs. 3.6 +/- 0.8, respectively. Eight p53 missense mutations were identified in six patients with recurrence and two controls. One nonsense mutation was found in a patient with recurrence and one deletion in a control patient. Only a minority of TP53 overexpression cases could be explained by the presence of these p53 mutations. CONCLUSION: TP53 overexpression was significantly predictive for recurrent endometrial carcinoma, and mostly not correlated with p53 mutations. Concomitant low hMdm2 and P21(Waf1/Cip1) expression in tumors with overexpressed TP53 suggests a dysfunctional TP53-P21(Waf1/Cip1) pathway. PMID- 16271750 TI - Cisplatin plus gemcitabine in previously treated squamous cell carcinoma of the cervix: a phase II study of the Gynecologic Oncology Group. AB - OBJECTIVES: This trial was conducted to evaluate the safety and efficacy of cisplatin plus gemcitabine in previously treated squamous cell carcinoma of the cervix. SUBJECTS AND METHODS: All women had measurable histologically confirmed squamous cell cervical cancer and a GOG performance status less than or equal to 2. The women were to receive cisplatin at 30 mg/m(2) plus gemcitabine at 800 mg/m(2) day 1 and day 8 every 28 days. RESULTS: Between February 2001 and May 2002, 32 eligible patients were entered. All women had received prior chemotherapy and 29 had received radiation. Twenty patients received platinum previously twice. The median time from primary treatment to recurrence was 21 months, but the median time from last prior chemotherapy was less than 2 months. A second phase of accrual was not indicated per the established stopping rules. There were 7 (21.9%) partial responses and median response duration was 2.1 months. Twelve additional women (37.5%) had stable disease. Nine women (28.1%) had increasing disease. Median time to progression was 3.5 months. There were no treatment-related deaths. Six women had grade 4 neutropenia, three had grade 4 anemia, and two had grade 4 thrombocytopenia. Grade 4 gastrointestinal toxicity occurred in two women and grade 4 anorexia occurred in one. CONCLUSIONS: This study suggests modest activity for the gemcitabine plus cisplatin doublet in previously treated squamous cell carcinoma of the cervix. The objective response rate of 22% is comparable to that of other active agents and combinations tested in this setting. Toxicities were primarily hematologic and generally manageable with dose reductions. PMID- 16271751 TI - Anti-tumor activity of TRA-8 anti-death receptor 5 (DR5) monoclonal antibody in combination with chemotherapy and radiation therapy in a cervical cancer model. AB - OBJECTIVES: There is substantial evidence that tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) causes apoptosis via activation of death receptors 4 and 5 (DR4 and DR5). We sought to determine the therapeutic potential of TRA-8 (anti-DR5 monoclonal antibody) in combination with chemotherapy and radiation therapy in a cervical cancer model. METHODS: DR5 expression in 7 human cervical cancer cell lines was analyzed by indirect immunofluorescence using murine TRA-8 in combination with flow cytometry. Cell lines were treated with TRA 8 alone or in combination with cisplatin, topotecan, or radiation, and cytotoxicity assays were performed. Mice were inoculated with ME-180 cancer cells and treated with different combinations of therapy. Animals receiving antibody were injected intraperitoneally with 200 microg of TRA-8. Animals received 9 Gy 60Co radiation divided into 3 fractions and 3 intraperitoneal doses of cisplatin (6 mg/kg) 1 h before radiation. A similar experiment was performed using topotecan (2 mg/kg) as the chemotherapeutic agent. RESULTS: DR5 was expressed to a varying degree on the cervical cancer cell lines. Combination treatment with TRA-8 and chemotherapy or radiation resulted in synergistic cytotoxicity in vitro. In vivo, combination therapy with TRA-8, cisplatin, and radiation produced tumor growth inhibition that was significantly greater than the other groups. Similar results were seen in combination studies with topotecan. CONCLUSIONS: These data suggest that DR5 is a good target for activation of the apoptotic pathway. Monoclonal antibodies such as TRA-8 may play an important role in the development of an effective treatment strategy for patients with advanced cervical cancer. PMID- 16271752 TI - Expression of maspin in gestational trophoblastic disease. AB - BACKGROUND: Maspin is a tumor suppressor gene whose expression is altered in neoplasia and malignancies of many tissues. In the human placenta, the maspin gene is expressed in trophoblastic cells and might act as an inhibitory regulator of trophoblastic invasion. Hence, in gestational trophoblastic disease (GTD), where there is increased propensity for invasion in the trophoblastic tissue, we hypothesized that maspin expression would be decreased. The present study aimed at investigating the expression of maspin in GTD and its prognostic significance. METHODS: Using immunohistochemical staining, we firstly studied the expression of maspin in hydatidiform moles, with gestational age-matched normal first trimester placenta used as control. A total of 38 cases of hydatidiform moles were studied, including 20 complete moles (CM) and 18 partial moles (PM). Among them, 10 cases of the CM group and 8 cases of the PM group subsequently developed gestational trophoblastic neoplasia (GTN). Immunostaining was also performed on tissue from 4 cases of choriocarcinoma and 5 cases of placental site trophoblastic tumor. Reverse transcriptase-polymerase chain reaction (RT-PCR) was further performed on RNA extracted from 10 hydatidiform moles (5 with GTN and 5 without) and 6 normal first-trimester placentae. RESULTS: In all tissue sections, nuclear expression of immunostaining signal was demonstrated, mainly in the cytotrophoblasts. The percentage of trophoblastic nuclei stained in both complete and partial moles was significantly lower than that in normal first-trimester placenta (P < 0.001). However, there was no significant difference in immunostaining between complete and partial moles (P > 0.05). There was also significantly lower expression of maspin in those cases subsequently developing GTN than those which did not (P = 0.01). Immunostaining on choriocarcinoma and placental site trophoblastic tumor showed reduced expression of maspin in all the tumor cells. Reverse transcriptase polymerase chain reaction revealed that the expression of maspin was consistently down-regulated in all the hydatidiform mole samples. CONCLUSIONS: Our results suggest that there is down-regulated expression of maspin in gestational trophoblastic diseases, and the down-regulation is more prominent in cases developing gestational trophoblastic neoplasia. This may play a role with prognostic significance in the pathogenesis and malignant transformation of hydatidiform moles. PMID- 16271753 TI - Trends in smoking by birth cohorts born between 1900 and 1977 in Japan. AB - BACKGROUND: The present study aimed to elucidate the changing patterns of smoking among successive birth cohorts in Japan. METHODS: Birth-cohort-specific smoking prevalence was estimated for birth cohorts born from 1900 to 1952, using data pooled from four prospective studies (242,330 men and 274,075 women), and for birth cohorts born from 1925 to 1977, using National Nutrition Survey data. RESULTS: For men, two peaks were observed in smoking prevalence, in the 1925 and late-1950s birth cohorts, while a trough was observed for the 1938 birth cohort. For women, ever smoking prevalence was lowest among the 1930s birth cohorts. After the female 1940s birth cohorts, no peak was observed until the end of our observations, the 1970s birth cohorts. Although Japanese women have historically tended to start smoking at later ages, recently, smoking habits have widely expanded among females in young birth cohorts. CONCLUSIONS: Smoking trends in Japanese men and women vary by birth cohorts. Smoking cessation should continue to be strongly promoted among men, although the younger generation has widely adopted a nonsmoking lifestyle. For women, efforts for preventing the onset of smoking, while necessary among the younger generation, should even be enhanced among middle-aged women. PMID- 16271754 TI - Performance of ten inbred mouse strains following assisted reproductive technologies (ARTs). AB - Superovulation, in vitro fertilization, embryo cryopreservation, and embryo transfer are assisted reproductive technologies (ARTs) widely used in laboratory mice. Inbred strains of mice have inherent genetic differences that cause them to respond differently to these technologies. Knowing how common inbred strains will perform when used for ARTs will ensure the most efficient use of mice, time, and resources. In this study, we characterized the ability of 10 inbred strains: 129S1/SvImJ, A/J, BALB/cJ, BALB/cByJ, C3H/HeJ, C57BL/6J, DBA/2J, FVB/NJ, NOD/LtJ, and SJL/J to superovulate, fertilize in vitro, and produce live pups subsequent to embryo transfer. Three-week-old female mice were superovulated using eCG (5.0 IU) and hCG (5.0 IU). The resulting oocytes were fertilized in vitro in human tubal fluid medium with spermatozoa of the same strain. The following day, two cell embryos were either transferred into pseudopregnant recipient females or cryopreserved. The cryopreserved embryos were later thawed and transferred into pseudopregnant recipient females. Differences in response to superovulation, fertilization, and number of live born produced after embryo transfer were observed between strains, substantiating the influence of genetic variability on ARTs. The response to the superovulation treatment varied among strains and ranged from 5+/-1(A/J) to 40+/-3 (129S1/SvImJ) normal oocytes per female. The average proportion of oocytes that fertilized ranged among strains from 24% (129S1/SvImJ) to 93% (DBA/2J and A/J). The average proportion of two-cell embryos that were transferred into recipient females and subsequently developed into live pups varied from 5% (A/J) to 53% (C57BL/6J) for fresh embryos and from 18% (BALB/cByJ) to 45% (129S1/SvImJ) for thawed embryos. PMID- 16271755 TI - Dysprosodic speech following basal ganglia insult: toward a conceptual framework for the study of the cerebral representation of prosody. AB - Progress in understanding brain/behavior relationships in adult-acquired dysprosody has led to models of cortical hemispheric representation of prosodic processing based on functional (linguistic vs affective) or physical (timing vs pitch) parameters. These explanatory perspectives have not been reconciled, and also a number of neurobehavior syndromes that include dysprosody among their neurological signs have not yet been integrated. In addition to expanding the functional perspective on prosody, some of these syndromes have implicated a significant role of subcortical nuclei in prosodic competence. In this article, two patients with acquired dysprosodic speech following damage to basal ganglia nuclei were evaluated using behavioral, acoustic, cognitive, and radiographic approaches. Selective quantitative measures were performed on each individual's performance to provide detailed verification and clarification of clinical observations, and to test hypotheses regarding prosodic function. These studies, combined with a review of related clinical research findings, exemplify the value of a broader perspective on the neurobehavioral dysfunction underlying acquired adult dysprosodic speech, and lead to a new, proposed conceptual framework for the cerebral representation of prosody. PMID- 16271756 TI - The effect of real-time aging on the oxidation and wear of highly cross-linked UHMWPE acetabular liners. AB - Irradiation decreases the wear of ultra-high molecular weight polyethylene (UHMWPE) but generates residual free radicals, precursors to long-term oxidation. Melting or annealing is used in quenching free radicals. We hypothesized that irradiated and once-annealed UHMWPE would oxidize while irradiated and melted UHMWPE would not, and that the oxidation in the former would increase wear. Acetabular liners were real-time aged by immersion in an aqueous environment that closely mimicked the temperature and oxygen concentration of synovial fluid. After 95 weeks of real-time aging, once-annealed components were oxidized; the melted components were not. The wear rate of the real-time aged irradiated and once-annealed components was higher than the literature reported values of other contemporary highly cross-linked UHMWPEs. Single annealing after irradiation used with terminal gamma sterilization may adversely affect the long-term oxidative stability of UHMWPE components. PMID- 16271757 TI - Fracture length scales in human cortical bone: the necessity of nonlinear fracture models. AB - Recently published data for fracture in human humeral cortical bone are analyzed using cohesive-zone models to deal with the nonlinear processes of material failure. Such models represent the nonlinear deformation processes involved in fracture by cohesive tractions exerted by the failing material along a fracture process zone, rather than attributing all damage to a process occurring at a single point, as in conventional linear-elastic fracture mechanics (LEFM). The relationship between the tractions and the net displacement discontinuity across the process zone is hypothesized to be a material property for bone. To test this hypothesis, the cohesive law was evaluated by analyzing published load vs. load point displacement data from one laboratory; the calibrated law was then used to predict similar data taken for a different source of bone using a different specimen geometry in a different laboratory. Further model calculations are presented to illustrate more general characteristics of the nonlinear fracture of bone and to demonstrate in particular that LEFM is not internally consistent for all cases of interest. For example, the fracture toughness of bone deduced via LEFM from test data is not necessarily a material constant, but will take different values for different crack lengths and test configurations. LEFM is valid when the crack is much longer than a certain length scale, representative of the length of the process zone in the cohesive model, which for human cortical bone ranges from 3 to 10mm. Since naturally occurring bones and the specimens used to test them are not much larger than this dimension for most relevant orientations, it is apparent that only nonlinear fracture models can give an internally consistent account of their fracture. The cohesive law is thus a more complete representation of the mechanics of material failure than the single parameter fracture toughness and may therefore provide a superior measure of bone quality. The analysis of fracture data also requires proper representation of the approximately orthotropic elasticity of the bone specimen; if the specimen is incorrectly assumed to be isotropic, the initial measured compliance cannot be reproduced to within a factor of four and the fracture toughness deduced from the measured work of fracture will be overestimated by approximately 30%. PMID- 16271758 TI - Detailed biochemical characterization of human placental cystatin (HPC). AB - A low molecular weight thiol protease inhibitor (12,500) purified from human placenta has been characterized in detail. Human placental cystatin (HPC) was found to be stable in the pH range 3.0-9.0 and temperature stability was between 40 and 100 degrees C. It does not have any disulphide groups and carbohydrate content. There was no cross-reaction between anti-HPC serum and other purified cystatins like HMW kininogens isolated from sheep plasma and phytocystatins isolated from Phaseolus mungo. The kinetics of inhibition of HPC was studied with ficin and bromelain and the comparison was made with our already reported results with papain. The respective K(i) values obtained for ficin and bromelain are 8.4 x 10(-8) M and 9.5 x 10(-8) M, respectively, whereas the value for papain was 5.5 x 10(-8) M. The values of association constants (K(ass)) for ficin and bromelain were 2.9 x 10(3) and 8.6 x 10(2) M(-1) s(-1), respectively, however, the value for papain was 3.4 x 10(4) M(-1) s(-1), the respective dissociation constant values for ficin and bromelain were 2.6 x 10(-5) and 2.1 x 10(-5) s(-1), respectively, and the value obtained for papain was 2.3 x 10(-5) s(-1). These kinetic parameters taken together along with t(1/2) values and IC(50) values imply that HPC binds more effectively to papain, then ficin and least with bromelain. Far-UV-CD analysis shows that HPC has 21.08% alpha-helical structure and significant amount of beta structure. Near-UV-CD spectra of HPC show positive peak at 280 nm indicating significant amount of tertiary interactions. The partial amino acid sequence analysis shows that HPC has highest sequence homology with chicken cystatin and Gly residue is present at position 11 rather than at conserved position 9, which has also been reported for human stefin A structure. The hydropathy plot of 1-30 amino acid residues shows that most amino acids of this stretch are present in the hydrophobic core of the protein. Owing to low molecular weight, absence of disulphide bonds and carbohydrate content HPC can be placed in type I cystatin family with some resemblance to chicken cystatin as shown by CD studies and amino acid sequence analysis. PMID- 16271760 TI - Inverse correlation between Flt3 and PU.1 expression in acute myeloblastic leukemias. AB - Over-expression of the Flt3 is prevalent in acute myeloblastic leukemia (AML), playing a role in leukemogenesis while decreased expression of PU.1 induces AML in mice model. Therefore, we speculated that there is an inverse relationship between these two factors. To clarify this, we measured the expression level of Flt3 and PU.1 in 24 primary AML specimens. As a result, there is a significant negative correlation between Flt3 and PU.1 (r=-0.43, p<0.05). Furthermore, we revealed that flt3 gene promoter is suppressed by the over-expression of PU.1, suggesting that PU.1 is a potential suppressor of flt3 gene promoter. PMID- 16271759 TI - Differential effects of N-peptidyl-O-acyl hydroxylamines on dynorphin-induced antinociception in the mouse capsaicin test. AB - In the capsaicin test, intrathecal (i.t.) dynorphins are antinociceptive. Cysteine protease inhibitors such as p-hydroxymercuribenzoate (PHMB) given i.t. augment and prolong their activity. The effect of two novel cysteine protease inhibitors, N-peptidyl-O-acyl hydroxylamines, on the antinociception induced by i.t. administered dynorphin A or dynorphin B has been investigated. When administered i.t. 5 min before the injection of capsaicin (800 ng) into the plantar surface of the hindpaw, dynorphin A (62.5-1000 pmol) or dynorphin B (0.5 4 nmol) produced a dose-dependent and significant antinociceptive effect. The effect of dynorphin A (1 nmol) and dynorphin B (4 nmol) disappeared completely within 180 and 60 min, respectively. PHMB (2 nmol) and Boc-Tyr-Gly-NHO-Bz (BYG Bz) (2 nmol) co-administered with dynorphin A or dynorphin B significantly prolonged antinociception induced by both. On the other hand, Z-Phe-Phe-NHO-Bz (ZFF-Bz) (1 and 2 nmol) only prolonged antinociception induced by dynorphin A. The results suggest that Z-Phe-Phe-NHO-Bz is an inhibitor of cysteine proteases preferring cleavage of dynorphin A, with less specificity towards dynorphin B in the mouse spinal cord. PMID- 16271761 TI - Food chain transfer of selenium in lentic and lotic habitats of a western Canadian watershed. AB - Selenium (Se) is an essential micronutrient, exhibiting a narrow margin between nutritionally optimal and potentially toxic concentrations. Egg-laying vertebrates at the top of aquatic food chains are most at risk in environments with elevated aqueous Se concentrations. The Elk River watershed in British Columbia, Canada receives effluents containing Se from five coal mine operations. This study tested three hypotheses that might account for higher Se concentrations in fish from lentic compared to lotic habitats in the watershed: (1) enhanced uptake by aquatic primary producers, (2) longer food chain length, or (3) greater food web accumulation through sediment-detrital pathways. Stable isotope and Se concentration data demonstrated that Se concentrations in aquatic primary producers and food chain lengths were comparable in lentic and lotic habitats. Enhanced formation of organoselenium and subsequent uptake and cycling via sediment detrital pathways likely account for higher fish tissue Se concentrations in lentic than in lotic areas. PMID- 16271762 TI - "Protoplasm...is soft wax in our hands": Paul Kammerer and the art of biological transformation. AB - Paul Kammerer's career ended in scandal in 1926 over tampering with his evidence for "Lamarckian" evolution--the infamous midwife toad. But although Kammerer's conclusions proved false, his evidence was probably genuine. In any case his arguments were not simply for Lamarckism and against Darwinism, as the theories are understood today. If we look beyond the scandal, the Kammerer story shows us a great deal about early 20th-century biology: the range of new ideas about heredity and variation, competing theories of biological and cultural evolution and their applications in eugenics, new kinds of laboratories and professional roles for biologists, and changing standards for documenting experimental results. PMID- 16271763 TI - Combat science: the emergence of Operational Research in World War II. AB - World War II became known as the "wizard war" because the cycles of developing countermeasures and counter-countermeasures to the weapons deployed by all sides drove rapid technological change. However, technological innovation was not the only contribution scientists made to the war effort. Through Operational Research (OR)--the scientific scrutiny of new weapons, their deployment and relative efficiency--scientists also influenced how warfare itself was conducted. This new scientific field emerged in the UK, where it helped to tighten the defense against the Luftwaffe. It quickly spread to other aspects of the military machine, improving both antisubmarine campaigns and bombing strategy. But although this analytical approach to warfare offered military commanders a factual basis on which to base difficult decisions and deal with tactical and strategic uncertainty, it was not without controversy. Indeed, several recommendations that came out of OR sparked disputes over the allocation of resources and strategic priorities. PMID- 16271764 TI - The Lysenko effect: undermining the autonomy of science. AB - The "Lysenko affair", which lasted roughly from the mid-1930s to the mid-1960s, was the big scandal of 20th-century science: a classic example of how politics can corrupt and undermine its rational basis. Under Stalin's leadership the Soviet Government suppressed genuine genetics and other sound biology, with devastating consequences for agriculture and health. The worst example of this occurred in August 1948 when the Politburo outlawed the teaching of and research into classical Mendelian genetics. There is broad agreement that this case offers a stark warning against politicians interfering with science. But what, precisely, is this interference that we are being warned about? Whereas the fate of genetics in Soviet Russia was a clear-cut example of direct suppression, there were also other less obvious ways in which politics subverted the scientific process. This indirect interference with science is a persistent feature of modern politics that we need to be on the lookout for. PMID- 16271765 TI - Claiming Copernicus. AB - The reputations of scientific heroes shift constantly, modified by politicians as well as by historians. Now that the Scientific Revolution has been reappraised, Nicolas Copernicus is portrayed as a friend of the Catholic Church rather than a scientific martyr. As a German-speaking Pole he has been claimed as a figure of national historical importance by both Germany and Poland, and since the early 20th century has been an important symbol of Polish independence. PMID- 16271766 TI - Electrical futures past. AB - Futurist writing about technology emerged in the late 19th century at the same time as new kinds of electrical technology were making utopian futures seem practically attainable. Electrical writers and novelists alike thus borrowed from the popular "science" fiction of Jules Verne, Edward Bellamy and others to try to create self-fulfilling prophecies of a future in which electrical gadgets and machines met all major practical needs of civilization. To the extent that many parts of our world are populated by the hardware that they forecast, they succeeded in their goal. PMID- 16271767 TI - HLA genes in Madeira Island (Portugal) inferred from sequence-based typing: footprints from different origins. AB - Human leukocyte antigen (HLA)-A, HLA-B, and HLA-DRB1 polymorphisms were examined in Madeira Island populations. The data was obtained at high-resolution level, using sequence-based typing (SBT). The most frequent alleles at each loci were: A*020101 (24.6%), B*5101 (9.7%), B*440201 (9.2%), and DRB1*070101 (15.7%). The predominant three-loci haplotypes in Madeira were A*020101-B*510101-DRB1*130101 (2.7%) and A*010101-B*0801-DRB1*030101 (2.4%), previously found in north and central Portugal. The present study corroborates historical sources and other genetic studies that say Madeira were populated not only by Europeans, mostly Portuguese, but also sub-Saharan Africans due to slave trade. Comparison with other populations shows that Madeira experienced a stronger African influence due to slave trade than Portugal mainland and even the Azores archipelago. Despite this African genetic input, haplotype and allele frequencies were predominantly from European origin, mostly common to mainland Portugal. PMID- 16271768 TI - cDNA cloning and phylogenetic analysis of the sixth complement component in rainbow trout. AB - The sixth complement protein (C6) is an essential component of the membrane attack complex (MAC); the end product of the lytic pathway of complement activation. The MAC complex constitutes a supramolecular assembly containing the five precursor proteins C5b, C6, C7, C8, and C9. Once assembled on the target surface it forms transmembrane channels that cause membrane damage and cytolysis of complement-opsonized pathogens. Besides mediating direct pathogen elimination, exposure of cells to sublytic doses of MAC can trigger diverse cellular responses such as, cell activation, induction of apoptosis, cell cycle re-entry and proliferation in various biological settings. The terminal complement components (C6-C9) are structurally related proteins, differing in size and complexity. In order to study their evolution, we report here the cloning and molecular characterization of C6 component in rainbow trout. The deduced amino acid sequence of trout C6 exhibits 55 and 44% identity with zebra fish and human orthologs, respectively. The 'domain' architecture of trout C6 resembles that of mammalian counterparts, and the cysteine backbone is also conserved. Finally, trout C6 gene appears to exist as a single copy in the trout genome, and is expressed in a wide range of trout tissues. PMID- 16271769 TI - Follow-up of patients affected by manganese-induced Parkinsonism after treatment with CaNa2EDTA. AB - In the period of 1998-2004, seven workers affected by manganese-induced Parkinsonism were diagnosed, studied and treated with CaNa2EDTA at our Occupational Health Ward. Biological markers, as well as magnetic resonance imaging and clinical examinations, were used to assess the disease trend. Those workers still employed were immediately removed from exposure. Our results seem to confirm that very good clinical, biological and neuroradiological results can be obtained by timely removal from exposure and chelating treatment, and that amelioration can persist in time. Manganism is, however, a severe condition that can also progress independent of further exposure. Therefore, chelating treatment can be a great aid in overt manganism, but particular attention must be paid to primary prevention, as this disease should now be totally preventable and definitely merits eradication. PMID- 16271770 TI - Rapid detection and quantitation of viral hemorrhagic septicemia virus in experimentally challenged rainbow trout by real-time RT-PCR. AB - A quantitative real-time RT-PCR (Q-RT-PCR) was developed to detect and determine the amount of viral hemorrhagic septicemia virus (VHSV) in organs of experimentally infected rainbow trout. Primers and TaqMan probes targeting the glycoprotein (G) and the nucleoprotein (N) genes of the virus were designed. The efficiency, linear range and detection limit of the Q-RT-PCR were assessed on cell cultured virus samples. VHSV N gene amplification was more efficient and more sensitive than the VHSV G amplicon. On cell culture grown virus, samples could be accurately assayed over a range of seven logs of infectious particles per reaction. To demonstrate the utility of Q-RT-PCR in vivo, bath infection trials were carried out and samples from fish spleen, kidney, liver and blood were harvested and tested for VHSV. Q-RT-PCR was a more reliable method than either conventional RT-PCR or the cell culture assay for virus diagnosis. Results of VHSV RNA detection in fish shortly after infection as well as on asymptomatic fish several weeks after experimental challenge are presented here. This is the first report showing the utility of Q-RT-PCR for VHSV detection and quantitation both in vitro and in vivo. The suitability of this method to test the efficacy of antiviral treatments is also discussed. PMID- 16271771 TI - A genotypic assay for the amplification and sequencing of gag and protease from diverse human immunodeficiency virus type 1 group M subtypes. AB - In human immunodeficiency virus type 1 (HIV-1), an interaction exists between the in vivo evolution of Gag protein and protease to escape from antiretroviral drug selective pressure. Therefore, it was decided to develop a genotypic assay for the amplification and sequencing of HIV-1 gag and protease. As the HIV-1 pandemic is characterised by a large genetic diversity, the assay developed was evaluated on a panel of 28 genetically divergent samples belonging to the following subtypes A1, B, C, D, F1, F2, G, H, J, CRF01-AE, CRF02-AG and CRF13-cpx. The assay displayed a detection limit ranging between 500 RNA copies/ml and 5000 RNA copies/ml plasma. Full-length sequences could be obtained for 25 samples. The population sequences of the three other samples lacked a part of the sequence because of heterogeneous signal, probably due to the presence of quasi-species with insertions/deletions of a different length. PMID- 16271772 TI - Protective effect of the acyclic nucleoside phosphonate tenofovir toward human T cell leukemia/lymphotropic virus type 1 infection of human peripheral blood mononuclear cells in vitro. AB - 9-(R)-[(2-Phosphonomethoxy)propyls]adenine (tenofovir), is an acyclic nucleoside phosphonate known to inhibit HIV replication in vitro and to reduce viremia in HIV-infected patients. Here we have investigated whether tenofovir is able to protect peripheral blood mononuclear cells (PBMCs) from healthy donors against human T-cell leukemia/lymphotropic virus type 1 (HTLV-1) infection in vitro. PBMCs were pre-treated with tenofovir and infected by exposure to an irradiated cell line chronically harbouring HTLV-1. Measurements of viral DNA, as well as viral gene and protein expression, at 4 weeks after infection, revealed that tenofovir at concentrations of 1 microM and higher completely protected PBMCs against HTLV-1; lower concentrations did not fully prevent HTLV-1 infection of the cultures. Nevertheless, in the long term, cell growth of infected PBMCs was inhibited in vitro even by 0.1 microM tenofovir. In addition, tenofovir directly inhibited HTLV-1 reverse transcriptase activity, in a cell-free assay that utilizes a crude preparation from HTLV-1 viral particles as a source of the enzyme. The selectivity index of tenofovir for HTLV-1, was about four times higher than that of azidothymidine. Taken together our results strongly encourage further studies to investigate the real impact of tenofovir towards HTLV-1 infection. PMID- 16271773 TI - Anxiety-like behaviors in pre-pubertal rats of the Flinders Sensitive Line (FSL) and Wistar-Kyoto (WKY) animal models of depression. AB - Animal models have been used in understanding the neuro-biological basis of depression and predicting successful treatment strategies. The current study focused on two genetic models of depression, the Flinder's Sensitive Line (FSL) and Wister-Kyoto (WKY). Our laboratory showed depressive symptomatology in pre pubertal WKY and FSL rats, and the current study focused on the strains' anxiety like traits. Since human depression-anxiety comorbidity is very common at young ages, it is essential to establish whether FSL and WKY pre-pubertal rats also exhibit such comorbidity. In addition, the effect of different rearing environments was studied using a mild chronic-stress condition (limiting available bedding between post-natal days 2-9). Two well-validated tests of anxiety, the open-field and elevated plus-maze, were used on 40-day-old pups. FSL pups exhibited lower anxiety-like behavior when compared to controls, in traditional open-field and plus-maze measures. A different pattern was observed in the WKY strain, which exhibited heightened anxiety-like behaviours in the FSL strain and affecting WKY's body-weight. Overall, the findings indicate differential expression of anxiety in pre-pubertal rats belonging to the 'depressed' strains, suggesting that these strains may be suitable for modelling different sub-groups of depression at young ages. PMID- 16271774 TI - Intestinal ischemia-reperfusion injury made leptin decreased. AB - To explore the role and the rule of leptin levels in severe traumatism, an ischemia-reperfusion injury model was established to observe change of leptin levels, and platelet activating factor, noradrenaline, lipopolysaccharide, and endothelin-1 were utilized to induce vascular endothelial cells. Leptin concentrations in serum and supernatant were detected by murine and human leptin radioimmunoassay. The results showed that the first serum leptin level significantly decreased after an injury of 60 min ischemia and 30 min reperfusion versus pre-experimental serum values, and leptin level in serum showed a variational trend to increase as reperfusion time extended; the second, supernatant leptin level significantly decreased after PAF and ET-1 treatments of 6 and 24 h versus the control group. It can be concluded that leptin maybe an inflammatory cytokine to play a protection role in acute inflammation and traumatism. PMID- 16271775 TI - Oral hygiene evaluation for effective oral care in preventing pneumonia in dentate elderly. AB - The purpose of this study was to establish criteria for the visual evaluation of oral hygiene by analyzing the relationship between status of oral hygiene and number of oral bacteria in saliva for use in predicting the development of pneumonia. A total of 145 Japanese people of advanced age living in nursing homes were enrolled in the study. We evaluated the Dental Plaque Index (DPI) and Tongue Plaque Index (TPI) as simple measures of status of oral hygiene. We also determined the number of viable microorganisms in the saliva of each subject. The relationship between the status of oral hygiene and episodes of pneumonia was investigated over a period of one year. Dentate patients with poor oral hygiene as indicated by their DPI and TPI scores demonstrated significantly higher salivary bacterial counts than those with a good score for oral hygiene (p<0.01 and p<0.05, respectively). Both the number of febrile days was significantly higher (p=0.0012), and number of patients developing pneumonia larger (p<0.01) in dentate patients with DPI-based poor scores than those with DPI-based good scores. These results demonstrate a significant positive correlation between salivary bacteria and visual evaluation of oral hygiene in dentate patients according to number of febrile days and development of pneumonia. PMID- 16271776 TI - Visualization of coronary artery anomalies by contrast-enhanced multi-detector row spiral computed tomography. AB - BACKGROUND: We investigated the potential of 16-slice multi-detector row spiral CT (MDCT) with retrospective ECG-gating for evaluation of coronary artery anomalies. METHODS: A total of 35 patients (23 men, 12 women, 19-81 years) in whom anomalous coronary arteries had been detected by invasive coronary angiography underwent MDCT (Sensation 16, Siemens, Germany). During one breathhold, a contrast-enhanced (90 ml, 5 ml/s) volume data set of the heart was acquired (16x0.75mm collimation, gantry rotation 375 ms). Images were reconstructed with a slice thickness of 1.0 mm in 0.5-mm intervals using retrospective ECG-gated reconstruction. The 35 data sets and 80 data sets of patients with angiographically normal coronary anatomy were analysed in random order concerning the origin and course of the coronary arteries. The results were compared to invasive coronary angiography. RESULTS: All patients with coronary artery anomalies and all controls with normal coronary anatomy were identified by MDCT. In addition, the origins and course concerning their anatomical relationship to adjacent cardiac structures were visualized in all patients, including right-sided origin of the left main (n=10), left anterior descending coronary artery (n=4) or left circumflex coronary artery (n=10); left-sided origin of the right coronary artery (n=6); four coronary fistula to the pulmonary artery (two from the left anterior descending, one from the right coronary artery, one from the left main), and one fistula from the circumflex coronary artery to the left atrium CONCLUSIONS: The study demonstrates that MDCT is a reliable noninvasive technique to identify and define anomalous coronary arteries and their course. PMID- 16271777 TI - Erectile dysfunction in healthy subjects predicts reduced coronary flow velocity reserve. AB - BACKGROUND: Erectile dysfunction is associated with, and may be the first sign of coronary artery disease. We aimed to assess whether men with erectile dysfunction but without cardiovascular disease have reduced coronary flow reserve, as a sign of early coronary atherosclerosis. METHODS: We investigated 12 men aged 68-73 years with erectile dysfunction, and 12 age-matched controls. Erectile function was evaluated using the validated IIEF-5 questionnaire. A score < or = 18 (of 25) was defined as erectile dysfunction and > or = 21 was considered normal. Patients with neurological or psychological reasons for erectile dysfunction were excluded, as were patients with symptoms of or prescribed medication for cardiovascular disease, hypertension or diabetes. Coronary flow velocity reserve was measured non-invasively by Doppler in the left anterior descending artery, before and during adenosine infusion. RESULTS: Coronary flow velocity reserve was significantly reduced in subjects with erectile dysfunction: 2.36 versus 3.19; P=0.024. In logistic regression analysis, compared to control subjects, men with erectile dysfunction had significantly increased risk of reduced coronary flow velocity reserve (< or = 3.0): odds ratio 15.4, P=0.02. In multivariate analysis, adjusting for age, tobacco use, systolic blood pressure, heart rate and body mass index, erectile dysfunction was the only significant predictor of reduced coronary flow velocity reserve, P=0.016. CONCLUSIONS: Men with erectile dysfunction but without diabetes or clinical cardiovascular disease have early signs of coronary artery disease. Our findings suggest that a cardiac risk evaluation may be indicated in men with suspected vasculogenic erectile dysfunction, and these individuals should be considered for primary prevention measures regarding cardiovascular disease risk factors. PMID- 16271778 TI - Prediction of necessity for coronary artery revascularization by adenosine contrast-enhanced magnetic resonance imaging. AB - BACKGROUND: Assessing myocardial first-pass wash-in during pharmacological induced stress allows detection of perfusion deficits and indicates stenotic coronary arteries (CA). The aim of our study was to demonstrate clinical relevance of contrast-enhanced stress magnetic resonance imaging (CMR) by predicting necessity of CA intervention. METHODS: 738 patients with scheduled coronary angiography (CXA) were scanned in a 1.5 Tesla CMR scanner. After 3 min of adenosine infusion (140 microg/kg/min), first-pass kinetic of contrast agent was evaluated. Myocardial necrosis was visualized with "myocardial late enhancement (MLE)". Perfusion deficits were described as either "ischemia in viable myocardium", or "no relevant ischemia in viable myocardium" or as "ischemia in chronic myocardial infarction (CMI)" based on spatial and temporal extent of ischemia and of MLE. CXA was performed in all patients within 48 h after CMR and revascularization, if applicable, was performed. Angiograms were read by two independent and blinded investigators and matched with CMR findings. RESULTS: 539 patients (73%) showed "ischemia in viable myocardium" and revascularization was performed in 513 patients (95%). In 111 patients with "no relevant ischemia in viable myocardium", revascularization was performed in only 5 patients (5%). In 88 patients classified as "ischemia in CMI", revascularization was performed in 14 patients (16%). Positive predictive value of CMR for CA intervention was 0.95, negative predictive value was 0.89, sensitivity was 0.96, and specificity was 0.87. CONCLUSION: CMR allows clinical useful prediction of relevant CA disease with need for revascularization prior to CXA and may be used as non-invasive test for myocardial ischemia and viability to guide further therapy. PMID- 16271780 TI - The prevalence of concomitant hypertension and hypercholesterolaemia in the general population. AB - OBJECTIVE: To study the joint prevalence of elevated blood pressure and raised total cholesterol in adult individuals and to estimate the multifactorial cardiovascular risk associated with this clustering. MATERIAL AND METHODS: Data were obtained through pooling of databases from large cross-sectional representative population-based studies carried out in Belgium. Prevalences of concurrent hypertension (systolic/diastolic blood pressure > or =140/90 mm Hg and/or under antihypertensive treatment) and hypercholesterolaemia (total cholesterol > or =190 mg/dl and/or using lipid-lowering drugs) were determined as well as the proportion of high risk individuals (according to SCORE) in this particular subgroup. RESULTS: In total, 16,300 men and 5075 women aged 35-74 years were included. The prevalences of hypertension and hypercholesterolaemia were, respectively, 41% and 84% in men and 31% and 82% in women. Both risk factors occurred simultaneously in 38% of men and 32% of women. In multivariate analysis, risk factor clustering was increasing with age and body mass index but appeared less prominent among smokers. Only 20% of all subjects having both risk factors were reaching a high multifactorial risk level of 5%. CONCLUSIONS: The high prevalence of co-existing hypertension and hypercholesterolaemia further accentuates the need for adequate risk factor management in this substantial subgroup of our population especially in younger subjects. PMID- 16271779 TI - Increased productivity of tumor necrosis factor-alpha in helper T cells in patients with systolic heart failure. AB - BACKGROUND AND AIMS: Whereas increased circulating proinflammatory cytokines, such as tumor necrosis factor-alpha (TNF-alpha), play an important role in heart failure, where and how TNF-alpha production is upregulated remains largely unknown. We studied the productivity of TNF-alpha in peripheral lymphocytes and underlying mechanisms in patients with heart failure. METHODS: Symptomatic NYHA II-IV patients with chronic heart failure with systolic dysfunction (n=39, aged 74+/-11, ejection fraction [EF]<==50%) were compared with asymptomatic NYHA I patients (n=18, aged 72+/-10, EF>50%) and normal subjects (n=15, aged 67+/-11). Lymphocyte subsets (CD3, CD4, and CD8) and intracellular production of TNF-alpha in peripheral leukocytes were quantified by immunofluorescent flow cytometry, and relationships between these parameters and circulating proinflammatory cytokines were analyzed. RESULTS: Subpopulation of TNF-alpha-producing CD4 was larger in NYHA II-IV patients (23.7% [18.0-28.6]) than in normal subjects (17.1% [6.5 19.5], p<0.05) and was correlated with plasma TNF-alpha levels (r=0.26, p<0.05), EF (r=-0.26, p<0.05), CD4/CD8 ratios (r=0.42, p<0.001), and subpopulation of TNF alpha-producing monocytes (r=0.47, p<0.0001). Plasma levels of soluble CD14 and interleukin-12 (IL-12) were significantly higher in NYHA II-IV patients than in normal subjects (1971 ng/mL [1740-2375] vs. 1607 ng/mL [1530-1930], p<0.01; and 121 pg/mL [62-230] vs. 62 pg/mL [54-99], p<0.05, respectively), and plasma IL-12 levels were correlated with plasma TNF-alpha levels (r=0.41, p<0.001). CONCLUSIONS: Increased productivity of TNF-alpha in helper T cells, associated with their dominance over cytotoxic T cells, may partially contribute to an increase in circulating TNF-alpha levels in heart failure. PMID- 16271781 TI - Woven coronary artery: a case report and review of literature. AB - Woven coronary artery is an extremely rare and clearly undefined coronary malformation. Up to now, very few cases have been reported. In this anomaly, epicardial coronary artery are branched into thin channels at any segment of the coronary artery and then these longitudinal twisted thin channels merge again as the main coronary lumen. This anomaly is regarded as a benign condition since there is completely normal blood flow after the distal segment of the abnormal coronary artery. In this case report, we present a 48-year-old male patient with a woven coronary artery anomaly in the circumflex artery and who had been followed up for 5 years. PMID- 16271782 TI - Stress stent fracture: is stent angioplasty really a safe therapeutic option in native aortic coarctation? AB - An 11-year-old boy was successfully treated by stent implantation for native aortic coarctation. At the 1-year control a severe re-coarctation was found at Doppler analysis and subsequent angiography revealed a transverse stent fracture. A stent-in-stent implantation was performed. Several hypotheses could explain this complication. Interventional cardiologists may pay more attention in following up these patients! PMID- 16271783 TI - Comparison of valuation methods used to generate the EQ-5D and the SF-6D value sets. AB - An interview study with 101 members of public compared the protocols used in valuation studies for EQ-5D (using ranking, visual analogue scale, and time trade off), and SF-6D (using ranking and standard gamble). Respondents were given one of the two protocols and asked to value four states each from EQ-5D and SF-6D. VAS scores suggest the narrower range of SF-6D values is partly attributable to the descriptive system; TTO values for milder states were higher than SG values; and the mean value for EQ-5D pits using TTO and SF-6D pits using SG were closer than across the two original valuation studies. PMID- 16271784 TI - Re: Hermann et al., low testosterone levels and quality of life. PMID- 16271785 TI - Dose-volume impact in high-dose-rate Iridium-192 brachytherapy as a boost to external beam radiotherapy for localized prostate cancer--a phase II study. AB - BACKGROUND AND PURPOSE: Evaluation of dose-volume-time-related factors in 64 patients treated with high-dose-rate brachytherapy (HDR-BT) as a boost to external beam radiotherapy (EBRT) for localized prostate cancer. PATIENTS AND METHODS: Clinical parameters were correlated with morbidity scores of the EPIC (Expanded Prostate Cancer Index) questionnaire. Median time after radiotherapy (HDR-BT up to 18 Gy in two fractions and EBRT up to a median dose of 50.4 Gy) was 1.5 and 3 years (first and second questionnaire). RESULTS: A significant impact of a urethra D1 exceeding 15 Gy in at least one HDR fraction concerning urinary morbidity and a rectum D1 exceeding 6 Gy to the rectal mucosa in the first and second HDR fraction concerning the rectal bleeding rate was found. A higher number of needles was associated with lower urinary and bowel scores after 1.5 years. A prostate length >4.8 cm and a longer duration of EBRT (independently of the dose) predisposed for lower urinary and bowel scores. In contrast to a urethra D1 > 15 Gy as an independent factor, a rectum D1 > 6 Gy per HDR fraction correlated with a higher number of needles and an increased prostate length. CONCLUSIONS: To minimize morbidity in HDR-BT for prostate cancer, a maximum dose to the urethra of 15 Gy and a maximum dose to the rectal mucosa of 6 Gy is advisable. Treatment- and patient-related factors have a major impact on toxicity. PMID- 16271787 TI - Potential of functional strains, isolated from traditional Maasai milk, as starters for the production of fermented milks. AB - The purpose of this research was the evaluation of technological features and of the ability of functional LAB strains with desirable sensory characteristics, to produce fermented milk. Eight strains of Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus paracasei and Lactococcus lactis, isolated from Maasai traditional fermented milk in Kenya and previously tested for their probiotic properties, were selected for this investigation. Technological features such as growth kinetics in fresh heat-treated whole milk medium and survival in the final product during storage at 4 degrees C, were studied. The strains Lb. acidophilus BFE 6,059, Lb. paracasei BFE 5,264 and Lc. lactis BFE 6,049 showed the best potential and were thus selected for use as starter cultures in further trials with the objective to improve their technological performance and to optimise the sensory features of fermented milk obtained. The effects of fat (F), non-fat milk solids (S) and fermentation temperature (T), modulated according to a Central Composite Design, on fermentation rates and viability losses during refrigerated storage of the chosen starters, and on product texture parameters, were studied. From the data analysis, it was possible to select optimum conditions for enhancing positive sensory traits of final products and for improving the survival of these potentially probiotic cultures. PMID- 16271786 TI - Delayed rectal and urinary symptomatology in patients treated for prostate cancer by radiotherapy with or without short term neo-adjuvant androgen deprivation. AB - BACKGROUND AND PURPOSE: To identify contributing factors to delayed rectal and urinary symptoms in a randomised trial comparing different durations of maximal androgen deprivation (MAD), given prior to radiotherapy, for locally advanced prostate cancer. PATIENTS AND METHODS: Between 1996 and 2000, 818 patients with stages T2b,c, 3 and 4 prostate cancer were entered into a trial comparing 0, 3 and 6 months of MAD prior to and during radiotherapy. Their delayed normal tissue effects were recorded by their treating doctors using standardised scales and by the patients using a self-assessment questionnaire regularly. Time to occurrence and prevalence data were analysed. RESULTS: Rectal and urinary symptom levels were observed to vary markedly over time in at least 80% of patients, with some indicating lasting resolution of symptoms. Prevalence rates were found to be substantially lower than actuarial probability rates. Baseline symptom levels and greatest acute symptom levels were both very powerful predictors. Obstructive lower urinary tract symptoms were noted to improve during the first 4 years after radiotherapy in approximately 60% of cases in each treatment arm. However, the treatment arm itself was not shown to influence these improvements in other univariate or multivariate analyses. MAD was shown to reduce both time to occurrence and prevalence of delayed proctopathic symptoms, but this effect was confirmed statistically in the 3 month treatment arm only. Multivariate models indicated that higher levels of haemoglobin prior to any treatment may in some way protect against delayed proctopathic symptoms. CONCLUSIONS: Prevalence data provide more clinically meaningful estimates of risk of delayed effects in normal tissues where assessment relies substantially on reported symptom levels. In these tissues consideration of the impact of baseline symptom levels and pathologies, and greatest acute symptom levels in analyses of delayed effects appears mandatory. Obstructive lower urinary symptoms improve over several years in the majority of patients treated for locally advanced prostate cancer by radiotherapy. Future research could address whether rectal toxicity is affected by initial haemoglobin levels and declines in it due to MAD. PMID- 16271789 TI - Lipolytic activity of Williopsis californica and Saccharomyces cerevisiae in extra virgin olive oil. AB - Inoculation trials performed with three strains of yeasts, isolated from extra virgin olive oil, Williopsis californica 1,639, Saccharomyces cerevisiae 1,525 and Candida boidinii 1,638, demonstrated that some yeast can lower the quality of the oil during storage. Laboratory tests highlighted a substantial increase in the total diglycerides and free fatty acids in the samples of oil inoculated with the lipase-producing strains of yeasts, W. californica 1,639 and S. cerevisiae 1,525, while in the samples of oil inoculated with the lipase-negative strain C. boidinii 1,638 no differences were found in respect to the uninoculated control. The acidity of the extra virgin olive oil, inoculated with the lipase-producing strains W. californica 1,639 and S. cerevisiae 1,525, during 2 weeks of incubation at 30 degrees C increased respectively from 0.62% to 1.50 and 1.62%, exceeding the limit of 0.8% established by current regulations for this commercial category of olive oil, while in the oil inoculated with the lipase negative strain and in the uninoculated control, the acidity remained constant throughout. Furthermore, the two strains of lipase-producing yeasts also increased the concentration of the 1.3-diglyceride isomer in the oil lowering the values of the total 1.2-diglycerides/total 1.3-diglycerides ratio considered to be an important index of quality for an extra virgin olive oil. The lipolytic activity of lipase-producing strains W. californica 1,639 and S. cerevisiae 1,525 showed an optimum pH of 6 and 7.5 and an optimum temperature of 20 degrees C and 30 degrees C respectively. Nevertheless, the lipolytic activity was negatively influenced by glucose and polyphenols when the concentration was higher than 0.25% and 0.4% (wt/vol) respectively. PMID- 16271788 TI - Streptococcus parauberis associated with modified atmosphere packaged broiler meat products and air samples from a poultry meat processing plant. AB - Lactic acid bacteria (LAB) isolated from marinated or non-marinated, modified atmosphere packaged (MAP) broiler leg products and air samples of a large-scale broiler meat processing plant were identified and analyzed for their phenotypic properties. Previously, these strains had been found to be coccal LAB. However, the use of a 16 and 23S rRNA gene RFLP database had not resulted in species identification because none of the typically meat-associated LAB type strains had clustered together with these strains in the numerical analysis of the RFLP patterns. To establish the taxonomic position of these isolates, 16S rRNA gene sequence analysis, numerical analysis of ribopatterns, and DNA-DNA hybridization experiments were done. The 16S rRNA gene sequences of three isolates possessed the highest similarities (over 99%) with the sequence of S. parauberis type strain. However, in the numerical analysis of HindIII ribopatterns, the type strain did not cluster together with these isolates. Reassociation values between S. parauberis type or reference strain and the strains studied varied from 82 to 97%, confirming that these strains belong to S. parauberis. Unexpectedly, most of the broiler meat-originating strains studied for their phenotypical properties did not utilize lactose at all and the same strains fermented also galactose very weakly, properties considered atypical for S. parauberis. This is, to our knowledge, the first report of lactose negative S. parauberis strains and also the first report associating S. parauberis with broiler slaughter and meat products. PMID- 16271790 TI - Trehalose-6-phosphate synthase as an intrinsic selection marker for plant transformation. AB - Insertion of foreign DNA into plant genomes occurs randomly and with low frequency. Hence, a selectable marker is generally required to identify transgenic plants. Until now, all selection systems have been based on the use of non-plant genes, derived from microorganisms and usually conferring antibiotic or herbicide resistance. The use of microorganism-derived genes however has raised biosafety concerns. We have developed a novel selection system based on enhancing the expression of a plant-intrinsic gene and the use of a harmless selection agent. Selection takes advantage of the reduced glucose sensitivity of seedlings with enhanced expression of AtTPS1, a gene encoding trehalose-6-P synthase. As a result, transformants can be identified as developing green seedlings amongst the background of small, pale non-transformed plantlets on high glucose medium. In addition, vegetative regeneration of tobacco leaf explants is very sensitive to high external glucose. Overexpression of AtTPS1 in tobacco allows selecting glucose insensitive transgenic shoots. PMID- 16271791 TI - A fluorescent antibiotic resistance marker for rapid production of transgenic rice plants. AB - Blasticidin S (BS) is an aminoacylnucleoside antibiotic used for the control of rice blast disease. To establish a new cereal transformation system, we constructed a visual marker gene designated gfbsd, encoding an enhanced green fluorescent protein (EGFP) fused to the N-terminus of BS deaminase (BSD). It was cloned into a monocot expression vector and introduced into rice (Oryza sativa L. cv. Nipponbare) calluses by microprojectile bombardment. Three to five weeks after the bombardment, multicellular clusters emitting bright-green EGFP fluorescence were obtained with 10 microg/ml BS, which is not sufficient to completely inhibit the growth of non-transformed tissues. Fluorescent sectors (approximately 2mm in diameter) excised from the calluses regenerated into transgenic plantlets (approximately 10 cm in height) as early as 51 (average 77+/ 11) days after the bombardment. The visual antibiotic selection was more efficient and required less time than the bialaphos selection with bar. In addition, the small size (1.1 kb) of gfbsd is preferable for construction of transformation vectors. This new marker gene will make a significant contribution in molecular genetic studies of rice plants. PMID- 16271792 TI - Present and future of cell-penetrating peptide mediated delivery systems: "is the Trojan horse too wild to go only to Troy?". AB - During the last decade, small peptides (10 to 15 amino acids) derived from the HIV-1 Tat protein and from the drosophila Antennapedia homeodomain have been used to internalize various types of molecules into the cells. The way these peptides enter cells is still under investigation and the object of strong controversy. The main discussions rely on whether these peptides are internalized or not in an energy-independent fashion, and, depending on the situation, whether they follow one pathway instead of another. At present, we find in the literature a very large number of data with, at times, some contradictory results. Indeed the diversity of employed peptide sequences, the cell type used, the attachment or not of a cargo molecule, the chemical nature of this cargo itself, and the followed protocol during the experimental process do not simplify the comparison and hence final conclusions about the mechanism of cell entry. However, one common feature emerges with these cell-penetrating peptides: most of them do not show any cell specificity. Despite their demonstrated efficiency in delivering biologically active molecules in in vitro experiments, their use for a therapeutic application in vivo has been the object of a relatively little number of studies, probably because of the quite important amounts of CPP-cargo that needs to be prepared for an accurate and complete in vivo study, but more likely, because of the massive spreading of the cargo all around the body. However, it appears from recent studies that an increased targeting ability of these CPPs is possible, making the use of CPP mediated delivery compatible with an in vivo therapeutic approach. PMID- 16271793 TI - Antitumor activity of poly(ethylene glycol)-camptothecin conjugate: the inhibition of tumor growth in vivo. AB - Antitumor effect of poly(ethylene glycol)-camptothecin conjugate (PEG-CPT) was studied in the nude mouse model of human colon cancer xenografts. The animals were treated intravenously with 15 mg/kg of camptothecin (CPT) or PEG-CPT conjugate at equivalent CPT dose. Antitumor activity, apoptosis induction and caspase-dependent signaling pathways were studied 12, 24, 48 and 96 h after single injection. In addition, pharmacokinetics, tumor distribution and accumulation of PEG polymer labeled with green fluorescence protein (GFP) were studied. The data obtained showed that the conjugation of low molecular weight anticancer drug CPT with low solubility to high molecular weight water-soluble PEG polymer provides several advantages over the native drug. First, the conjugation improves drug pharmacokinetics in the blood and tumor. Second, such conjugation provides passive tumor targeting by the Enhanced Permeability and Retention (EPR) effect, increasing drug concentration in the tumor. Third, the coupling increases the bioavailability of CPT, induces apoptosis in tumor and, therefore, enhances anticancer activity of PEG-CPT. Thus, the use of macromolecular conjugate provided passive tumor targeting of the drug, improved pharmacokinetics and increased the stability of the drug during circulation. It offered better uptake by the targeted tumor cells and substantially enhanced apoptosis and antitumor activity of the conjugated drug in the tumor and decreased apoptosis in liver and kidney as compared with the native drug. All these characteristics make PEG-CPT conjugate an attractive anticancer drug for the effective chemotherapy of solid tumors. PMID- 16271794 TI - Hepatitis C minimal residual viremia (MRV) detected by TMA at the end of Peg-IFN plus ribavirin therapy predicts post-treatment relapse. AB - BACKGROUND/AIMS: Around 15-25% of chronic hepatitis C patients treated with Peg IFN plus ribavirin become HCV-RNA negative by PCR during therapy but relapse after its withdrawal. We investigated whether minimal residual viremia (MRV) might be detected in these cases by Transcription-Mediated Amplification (TMA). METHODS: Two hundred and ninety-two consecutive patients (143 HCV-1, 82 HCV-2, 56 HCV-3 and 11 HCV-4) were prospectively treated with a standard schedule of Peg IFNalpha 2b plus ribavirin combination and end-of-therapy response was assessed by conventional PCR using 2 protocol serum samples obtained 6-8h before the last two scheduled weekly injections of Peg-IFN. PCR negative samples were re-tested by TMA and the results were then correlated with the virological outcome after therapy withdrawal. RESULTS: Among 208 patients who were repeatedly HCV-RNA negative by PCR at the end-of-therapy, 26 (12.5%) were found HCV-RNA positive by TMA. Twenty-two of them, (96%) were PCR-relapsers after therapy withdrawal, compared to only 14% of the 182 TMA negative patients (P<0.0001). This virological profile was more frequent in HCV-1 and HCV-3 infected patients and correlated with a slower virological response during therapy. CONCLUSIONS: At the end of Peg-IFN plus ribavirin therapy, TMA is superior to PCR in identifying patients with sustained HCV-RNA clearance. PMID- 16271795 TI - A pilot approach for quantitative assessment of liver fibrosis using ultrasound: preliminary results in 79 cases. AB - BACKGROUND/AIMS: Ultrasound is noninvasive and useful to evaluate liver disease despite its operator dependency. This pilot study was conducted to quantitatively assess liver fibrosis using ultrasound. METHODS: Fibrosis extraction ratios (FER) (fiber volume/total volume) of ultrasound and histological images of 8 autopsy specimens were compared. We also compared FER of ultrasound images from clinical patients (n=79) with histological fibrosis stages. RESULTS: In the autopsy study, FER correlation coefficient between histological images and ultrasound images was 0.992. Regarding clinical patients, there was sufficient evidence to indicate differences in the distributions of FER for each fibrosis stage (Kruskal-Wallis test P<0.0001). With FER cut-off to distinguish > or =F2 from F0 and F1 defined as mean plus standard deviation of F0 and F1, sensitivity, specificity, positive predictive value, negative predictive value, and likelihood ratio were 62, 75, 78, 57%, and 2.47, respectively. Regarding HCV cohort (n=44), they were 55, 87, 89, 50%, and 4.14, respectively. Areas under receiver operating characteristic curves were 0.78, 0.79, 0.83 and 0.83 for > or =F1, > or =F2, > or =F3 and =F4, respectively. Regarding HCV cohort, they were 0.74, 0.71, 0.79 for > or =F2, > or =3 and =4, respectively. CONCLUSIONS: The FER method has great potential for diagnosing liver fibrosis using ultrasound. PMID- 16271796 TI - Limited iron export by hepatocytes contributes to hepatic iron-loading in the Hfe knockout mouse. AB - BACKGROUND/AIMS: In hereditary hemochromatosis, iron-loading of hepatocytes is associated with increased iron uptake while little is known about iron release. This study aims to characterise iron release and ferroportin expression by Hfe knockout hepatocytes to determine if they contribute to iron overload in haemochromatosis. METHODS: Iron release by hepatocytes from Hfe knockout, non iron-loaded and iron-loaded wild-type mice was measured after incubation with nontransferrin-bound iron as iron-citrate. RESULTS: Iron release and ferroportin expression by hepatocytes from Hfe knockout, non-iron-loaded and in vivo iron loaded wild-type mice were similar although, nontransferrin-bound iron uptake was significantly increased in Hfe knockout hepatocytes and decreased in iron-loaded wild-type hepatocytes compared with non-iron-loaded wild-type cells. When expressed as a percentage of total iron uptake, iron release was decreased in Hfe knockout hepatocytes (4.6+/-0.7 versus 13.7+/-1.2%, P<0.0001) and increased in iron-loaded wild-type hepatocytes (29.5+/-0.5 versus 13.5+/-0.7%; P<0.0001) compared with wild-type hepatocytes. In contrast, in vitro iron-loading increased iron release and ferroportin expression by both Hfe knockout and wild-type hepatocytes. CONCLUSIONS: Hfe knockout hepatocytes accumulate iron as a result of limited iron export and enhanced iron uptake. The correlation between iron release and ferroportin expression suggests that iron export in hepatocytes is mediated by ferroportin. PMID- 16271800 TI - Microdialysis assessment of drug delivery systems for vitreoretinal targets. AB - Posterior segment drug delivery challenges inherent in the treatment of many sight-threatening diseases have become increasingly apparent. Therapeutic interventions for ocular diseases such as neovascular retinopathies, inflammatory and/or infectious diseases may involve drug delivery to vitreoretinal targets. An important part of successful therapeutic strategies for such diseases involves verification that efficacious concentrations of the pharmacological agent are achieved within relevant intraocular regions. Microdialysis has been effectively employed for characterizing intraocular disposition in both anterior and posterior segments, providing important documentation of successful drug delivery to desired targets. Recent papers that showcase the maturation in the model development of microdialysis approaches for estimating posterior segment pharmacokinetics and further validation of the methodology are described in this review. Special problems examined include anterior and posterior ocular clearance mechanisms, intraocular metabolism and active transport of drugs. PMID- 16271798 TI - Cocaine induction of ERK proteins in dorsal striatum of Fischer rats. AB - Cocaine is an addictive psychostimulant that induces fos and opioid gene expression by activating the dopamine receptors and the PKA pathways in dopamine D1 and a glutamate NMDA-dependent mechanisms in the striatum. In this study, we show that a single cocaine administration induces ERK phosphorylation in the caudate/putamen of Fischer rats. This increase in Phospho-ERK is diminished by pre-administration of SCH23390, or MK801 but not with pre-administration of eticlopride. Furthermore, this single cocaine administration does not alter the levels of phospho-CREB protein or CREB-DNA bindings in the caudate/putamen protein extracts but does increase phospho-Elk-1 protein levels in the same extracts. PMID- 16271801 TI - NMDA modulation of dopamine dynamics is diminished in the aged striatum: an in vivo voltametric study. AB - The technique of in vivo voltametry and a paired recording paradigm were employed to study the age-related changes in N-methyl-d-aspartate (NMDA) function in regulating the striatal dopaminergic transmission in male Sprague-Dawley rats. Microinjection of NMDA (100pmol) consistently elicited larger striatal dopamine (DA) overflows from young rats (3-4 months old) than from aged rats (27-28 months old). Furthermore, the rate of clearance (T(c)) of the NMDA-evoked dopamine release was lower in the aged rats. Local application of dopamine evoked reversible electrochemical signals with similar amplitudes in both young and aged rats. However, T(c) was reduced and time course parameters were prolonged in the aged rats. While microejection of NMDA (1pmol) did not induce any dopamine overflow, simultaneous administration of NMDA and K(+) evoked larger dopamine releases than K(+) alone in the young striatum. Concomitant application of NMDA did not potentiate the K(+)-evoked dopamine release in the aged striatum. Taken together, with the reduced dopamine release in response to depolarizing stimuli, our in vivo electrochemical data suggest that age-related changes in NMDA function contribute to the impaired dopaminergic dynamics, including an attenuation of NMDA-evoked dopamine release and a diminished augmentation by K(+) of NMDA-induced dopamine release during the normal aging process. PMID- 16271802 TI - Expression of carbonic anhydrases II, IV, VII, VIII and XII in rat brain after kainic acid induced status epilepticus. AB - Carbonic anhydrases (CAs) are important enzymes in the central nervous system (CNS), where they participate in regulating cerebrospinal fluid (CSF) secretion, blood-brain barrier and glial cell function. Using RT-PCR we found CA XII mRNA in rat and mouse brain. Cloning of rat CA XII revealed 94% homology with the mouse CA XII. To map the putative functional roles of different CAs, we studied the expression and localization of CA II, CA IV, CA VII, CA-related protein (CA-RP) VIII and CA XII mRNAs in rat brain after kainic acid induced epileptic seizures using Northern blot analysis and in situ hybridization. The expression of CA IV, CA VII and CA-RP VIII was somewhat similar: they were expressed in the cortex, hippocampus and midbrain structures and their expression did not change after the kainic acid treatment. The expression of CA II was concentrated in the white matter structures, which is in line with the preferential expression of CA II in the oligodendrocytes. High levels of CA II mRNA were also detected in the choroid plexus. Surprisingly, CA II was induced 3-12 h after seizures in the vulnerable CA1 region. CA XII was expressed in dentate granule cells, cortex and choroid plexus. Kainic acid stimulated CA XII expression throughout the cortical layer I. The observed hippocampal induction of CA II may indicate a pro-apoptotic and/or epileptogenic role of CA II after prolonged seizures. The physiological significance of the observed cortical induction of CA XII remains obscure. Cytosolic CA II is known to participate in CSF secretion, and the high expression of CA XII in the choroid plexus suggests an analogous role for this membrane bound isozyme. PMID- 16271803 TI - BioVision: an application for the automated image analysis of histological sections. AB - We describe a computer application, "BioVision", that can be trained to quickly and effectively classify and quantify user definable histological objects (e.g., senile plaques, neurofibrillary tangles) within single or double-labeled immunocytochemically stained sections. For a given image population, BioVision is interactively trained (in Independent User Mode) by an investigator to perform the desired classifications. This training yields a statistical model of the different types of objects occurring in the target image population. The resulting model can then be used (in Automated User Mode) to classify all objects in any image or images from the target population. BioVision simplifies the quantification of complex visual objects and improves inter-rater reliability. The program accomplishes classification in two major stages: pixel classification and blob classification. In pixel classification, each pixel is assigned to one of some number of substance classes, based on its chromatic properties and local context, reflecting basic histological distinctions of interest. In the blob classification phase, the image's pixels are first partitioned into "blobs": maximal connected sets of pixels assigned to the same substance class. Then, based on its size, shape, textural and contextual properties, each blob is assigned to a histological object class. A Bayesian classifier is used in each of the pixel and blob classification stages. We report several tests of BioVision. First, we applied BioVision to classify senile plaques and neurofibrillary tangles in several test cases of Alzheimer's brain immunostained for beta-amyloid and PHF-tau and compared the results to those produced by experienced investigators. BioVision was trained to classify Plaque-type blobs as either plaques or plaque-type nonentities, and tangle-type blobs as either tangles or tangle-type nonentities. BioVision classified the objects with an accuracy comparable to the trained investigator. Next, we applied BioVision to the task of counting all the tangles in hippocampal images from 22 Alzheimer's disease (AD) cases selected to span a broad range of dementia levels from the tissue repository of UC Irvine's Center for the study of Brain Aging and Dementia. The tangle counts produced by BioVision proved to be significantly better predictors of the cases' adjusted MMSE scores than any of tangle load, age at death, post mortem interval or the interval between the last MMSE score and death. PMID- 16271804 TI - Redox proteomics identification of oxidized proteins in Alzheimer's disease hippocampus and cerebellum: an approach to understand pathological and biochemical alterations in AD. AB - Alzheimer's disease (AD) is characterized by the presence of neurofibrillary tangles, senile plaques and loss of synapses. There is accumulating evidence that oxidative stress plays an important role in AD pathophysiology. Previous redox proteomics studies from our laboratory on AD inferior parietal lobule led to the identification of oxidatively modified proteins that were consistent with biochemical or pathological alterations in AD. The present study was focused on the identification of specific targets of protein oxidation in AD and control hippocampus and cerebellum using a redox proteomics approach. In AD hippocampus, peptidyl prolyl cis-trans isomerase, phosphoglycerate mutase 1, ubiquitin carboxyl terminal hydrolase 1, dihydropyrimidinase related protein-2 (DRP-2), carbonic anhydrase II, triose phosphate isomerase, alpha-enolase, and gamma-SNAP were identified as significantly oxidized protein with reduced enzyme activities relative to control hippocampus. In addition, no significant excessively oxidized protein spots were identified in cerebellum compared to control, consistent with the lack of pathology in this brain region in AD. The identification of oxidatively modified proteins in AD hippocampus was verified by immunochemical means. The identification of common oxidized proteins in different brain regions of AD brain suggests a potential role for these oxidized proteins and thereby oxidative stress in the pathogenesis of Alzheimer's disease. PMID- 16271805 TI - Amyloid beta peptide as a physiological modulator of neuronal 'A'-type K+ current. AB - Control of neuronal spiking patterns resides, in part, in the type and degree of expression of voltage-gated K(+) channel subunits. Previous studies have revealed that soluble forms of the Alzheimer's disease associated amyloid beta protein (Abeta) can increase the 'A'-type current in neurones. In this study, we define the molecular basis for this increase and show that endogenous production of Abeta is important in the modulation of Kv4.2 and Kv4.3 subunit expression in central neurones. A-type K(+) currents, and Kv4.2 and Kv4.3 subunit expression, were transiently increased in cerebellar granule neurones by the 1-40 and 1-42 forms of Abeta (100nM, 2-24h). Currents through recombinant Kv4.2 channels expressed in HEK293 cells were increased in a similar fashion to those through the native channels. Increases in 'A'-type current could be prevented by the use of cycloheximide and brefeldin A, indicating that protein expression and trafficking processes were altered by Abeta, rather than protein degredation. Endogenous Abeta production in cerebellar granule neurones was blocked using inhibitors of either gamma- or beta-secretase and resulted in decreased K(+) current. Crucially this could be prevented by co-application of exogenous Abeta (1nM), however, no change in Kv4.2 or Kv4.3 subunit expression occurred. These data show that Abeta is a modulator of Kv4 subunit expression in neurones at both the functional and the molecular level. Thus Abeta is not only involved in Alzheimer pathology, but is also an important physiological regulator of ion channel expression and hence neuronal excitability. PMID- 16271806 TI - Regionally specific atrophy of the corpus callosum in AD, MCI and cognitive complaints. AB - The goal of the present study was to determine if there are global or regionally specific decreases in callosal area in early Alzheimer's disease (AD) and mild cognitive impairment (MCI). In addition, this study examined the corpus callosum of healthy older adults who have subjective cognitive complaints (CC) but perform within normal limits on neuropsychological tests. We used a semi-automated procedure to examine the total and regional areas of the corpus callosum in 22 patients with early AD, 28 patients with amnestic MCI, 28 healthy older adults with cognitive complaints, and 50 demographically matched healthy controls (HC). The AD, MCI, and CC groups all showed a significant reduction of the posterior region (isthmus and splenium) relative to healthy controls. The AD group also had a significantly smaller overall callosum than the controls. The demonstration of callosal atrophy in older adults with cognitive complaints suggests that callosal changes occur very early in the dementing process, and that these earliest changes may be too subtle for detection by neuropsychological assessments, including memory tests. PMID- 16271807 TI - [Pathophysiology and therapeutic implications of left heart failure]. AB - PURPOSE: Heart failure is the ultimate step of most cardiovascular diseases. Its frequency increases regularly because of the progressive increase of life expectancy and better management of cardiovascular diseases. The prognosis is very poor (5-year mortality: 50%) as is quality of life; heart failure is a very costly disease. CURRENT KNOWLEDGE AND KEYS POINTS: Aims of treatment are improvement of symptoms and thus improvement of quality of life and increase of survival. The treatment systematically combines: general advice, dietary measures; medical treatment (with betablockers, ACE inhibitors and/or angiotensin II receptor antagonists, diuretics, in some cases aspirin or oral anticoagulants, digitalis and amiodarone) according to severity of heart failure, presence of congestion, aetiology, age; etiologic treatment if possible; treatment and prevention of precipitating and exacerbating factors. According to clinical and paraclinical features, one may propose cardiac multisite stimulation, cardiac surgery, physical stress training and cardiac transplantation. In order to decrease frequency of heart failure, prevention of cardiovascular diseases which lead to heart failure must be done as often is possible (hypertension, valvular heart disease, ischemic heart disease). FUTURE PROSPECTS AND PROJECTS: The future of the treatment of heart failure is the multidisciplinary management of heart failure (networks) led by hospital units specialized in heart failure. PMID- 16271808 TI - A recombinant 63-kDa form of Bacillus anthracis protective antigen produced in the yeast Saccharomyces cerevisiae provides protection in rabbit and primate inhalational challenge models of anthrax infection. AB - Infection by Bacillus anthracis is preventable by prophylactic vaccination with several naturally derived and recombinant vaccine preparations. Existing data suggests that protection is mediated by antibodies directed against the protective antigen (PA) component of the anthrax toxin complex. PA is an 83-kDa protein cleaved in vivo to yield a biologically active 63-kDa protein. In an effort to evaluate the potential of yeast as an expression system for the production of recombinant PA, and to determine if the yeast-purified rPA63 can protect from a lethal inhalational challenge, the sequence of the 63-kDa form of PA was codon-optimized and expressed in the yeast Saccharomyces cerevisiae. Highly purified rPA63 isolated from Saccharomyces under denaturing conditions demonstrated reduced biological activity in a macrophage-killing assay compared to non-denatured rPA83 purified from Escherichia coli. Rabbits and non-human primates (NHP) immunized with rPA63 and later challenged with a lethal dose of B. anthracis spores were generally protected from infection. These results indicate that epitopes present in the 63-kDa from of PA can protect rabbits and non-human primates from a lethal spore challenge, and further suggest that a fully functional rPA63 is not required in order to provide these epitopes. PMID- 16271810 TI - Duration of anticoagulation: decision making based on absolute risk. AB - We are often faced with the question as to the optimum duration of secondary prophylaxis with oral anticoagulants after an episode of venous thromboembolism. Theoretically if we know the recurrence rate, the case-fatality, the effectiveness of oral anticoagulant therapy, and the rate of fatal haemorrhage on treatment, we can calculate whether being on or off treatment is safest. Using these data and considering only the risk of death we would treat idiopathic deep vein thrombosis for six months. For those with DVT associated with a transient risk factor it would be reasonable to stop treatment after 3 months in those over 50 years old and we should certainly stop after 3 months in those over 70 years old. There are data to suggest that pulmonary embolism may have a higher case fatality than deep vein thrombosis if there is a recurrence. If these data were accepted most patients with idiopathic pulmonary embolism would get long-term treatment. We can use these models to modify our assessment if other factors such as antiphospholipid antibodies or cancer are present. PMID- 16271809 TI - Pathological Teres Major activation in patients with massive rotator cuff tears alters with pain relief and/or salvage surgery transfer. AB - BACKGROUND: Massive rotator cuff tears impose restraints on overhand arm functionality and are often accompanied by pain. After musculotendinous Teres Major transfer, overhand arm function is generally restored and pain is reduced. The assumed mechanical abduction insufficiency and Teres Major muscle function adaptation will be experimentally verified. METHODS: Principal Teres Major muscle activation (surface IEMG averaged over 3s) is recorded for 12-24 isometric and isotonic force directions perpendicular to the 60 degrees forward flexed humerus in three conditions: prior to surgery (n = 6 patients), prior to surgery and after subacromial anaesthetic (n = 6) and post-surgery (n = 3). Principal direction and on-, offset directions were estimated. FINDINGS: Teres Major activation adapts both to pathological and post surgery conditions: the normal activation during adduction changes into activation during forward flexion or abduction. Glenohumeral stabilisation, not abduction torque, seems to be the explanation for post surgery Teres Major transfer success. INTERPRETATIONS: The pathological absence of Supraspinatus and Infraspinatus forces during forward flexion result in increased upward glenohumeral instability. The superior translations are compensated for by Teres Major activity during forward flexion. This translation-'force' function conflicts with the adduction-generating rotation-'torque' function. This may explain the pain-induced reduction of arm elevation in these patients. Musculotendinous transfer solves the force-torque conflict by changing the moment arm of the Teres Major from adduction to abduction. Teres Major can now both compensate for the loss of Supraspinatus and Infraspinatus forces needed for glenohumeral stabilisation and contribute to forward flexion of the arm. PMID- 16271811 TI - Preliminary assessment of risk of ozone impacts to maize (Zea mays) in southern Africa. AB - Surface ozone concentrations in southern Africa exceed air quality guidelines set to protect agricultural crops. This paper addresses a knowledge gap by performing a preliminary assessment of potential ozone impacts on vegetation in southern African. Maize (Zea mays L.) is the receptor of interest in the main maize producing countries, i.e. South Africa, Zambia and Zimbabwe. Surface ozone concentrations are estimated for the growing season (October to April) using photochemical modelling. Hourly mean modelled ozone concentrations ranged between 19.7 and 31.2 ppb, while maximums range between 28.9 and 61.9 ppb, and are near 30 ppb over South Africa and Zambia, while in Zimbabwe, they exceed 40 ppb and translate into monthly AOT40 values of over 3,000 ppb h in five of the seven months of the growing season. This study suggests that surface ozone may pose a threat to agricultural production in southern African, particularly in Zimbabwe. PMID- 16271812 TI - Atmospheric concentrations and air-sea exchanges of nonylphenol, tertiary octylphenol and nonylphenol monoethoxylate in the North Sea. AB - Concentrations of nonylphenol isomers (NP), tertiary octylphenol (t-OP) and nonylphenol monoethoxylate isomers (NP1EO) have been simultaneously determined in the sea water and atmosphere of the North Sea. A decreasing concentration profile appeared following the distance increasing from the coast to the central part of the North Sea. Air-sea exchanges of t-OP and NP were estimated using the two-film resistance model based upon relative air-water concentrations and experimentally derived Henry's law constant. The average of air-sea exchange fluxes was -12+/-6 ng m(-2)day(-1) for t-OP and -39+/-19 ng m(-2)day(-1) for NP, which indicates a net deposition is occurring. These results suggest that the air-sea vapour exchange is an important process that intervenes in the mass balance of alkylphenols in the North Sea. PMID- 16271813 TI - Response of the Mediterranean sponge Chondrosia reniformis Nardo to copper pollution. AB - We examined the effects of exposure to copper pollution on the Atlanto Mediterranean sponge Chondrosia reniformis. We transplanted sponges from an unpolluted control area to a harbour with a moderately high concentration of copper and measured several biological sponge variables. No effect of this habitat was detected on sponge growth, shape, heat-shock protein expression or metal accumulation. However, a decrease in the clearance rate, an increase in the collagen/cell rate (due to a decrease in the cellular components) and a lower survival rate after 4 months of the sponges transplanted to the harbour was observed. We suggest that copper may alter the sponge physiology, by reducing pumping capacity, which may ultimately lead to sponge death. Consequently, copper pollution exerts strong negative effects on this organism. PMID- 16271814 TI - Regulating manure application discharges from concentrated animal feeding operations in the United States. AB - In the United States, reducing pollution from agriculture has received attention due to data suggesting that this is the leading source of impairment of many waterbodies. The federal government revised its regulations governing concentrated animal feeding operations (CAFOs) to enhance governmental oversight over sources of pollution. For the application of manure resulting in pollutant discharges, CAFOs need to implement nutrient management plans. A federal court affirmed the ability of the US federal government to oversee the application of manure from CAFOs that have discharges. Simultaneously, owners and operators of CAFOs who have implemented an appropriate nutrient management plan may forgo securing a permit if their discharges qualify under the agricultural stormwater discharge exemption. PMID- 16271815 TI - Meta-analysis of therapeutic relationship variables in youth and family therapy: the evidence for different relationship variables in the child and adolescent treatment outcome literature. AB - This meta-analysis examines associations between therapeutic relationship variables, and the extent to which they account for variability in treatment outcomes, in 49 youth treatment studies. Correlations between therapeutic relationship variables ranged from modest to strong. Among the best predictors of youth outcomes were counselor interpersonal skills, therapist direct influence skills, youth willingness to participate in treatment, parent willingness to participate in treatment, youth participation in treatment, and parent participation in treatment. Adequacy of current approaches to conceptualizing and measuring therapeutic relationship variables, such as the therapeutic alliance, in youth and family therapy is discussed. This paper represents the most comprehensive analysis of therapeutic relationship constructs in the youth treatment literature. PMID- 16271816 TI - Prehension movements in a patient (AC) with posterior parietal cortex damage and posterior callosal section. AB - Prehension movements of the right hand were recorded in a right-handed man (AC), with an injury to the left posterior parietal cortex (PPC) and with a section of the left half of the splenium. The kinematic analysis of AC's grasping movements in direct and perturbed conditions was compared to that of five control subjects. A novel effect in prehension was revealed--a hemispace effect--in healthy controls only. Movements to the left hemispace were faster, longer, and with a smaller grasp aperture; perturbation of both object position and distance resulted in the attenuation of the direction effect on movement time and the time to velocity peak, with a reverse pattern in the time to maximum grip aperture. Nevertheless, the correlation between transport velocity amplitude and grasp aperture remained stable in both perturbed and non-perturbed movements, reflecting the coordination between reaching and grasping in control subjects. In contrast, transport and grasp, as well as their coordination in both direct and perturbed conditions, were negatively affected by the PPC and splenium lesion in AC, suggesting that transport and grasp rely on two functionally identifiable subsystems. PMID- 16271817 TI - Behavioral and physiological findings of gender differences in global-local visual processing. AB - Hemispheric asymmetries in global-local visual processing are well-established, as are gender differences in cognition. Although hemispheric asymmetry presumably underlies gender differences in cognition, the literature on gender differences in global-local processing is sparse. We employed event related brain potential (ERP) recordings during performance of a global-local reaction time task to compare hemispheric asymmetries and processing biases in adult men (n=15) and women (n=15). Women responded more quickly to local targets while men did not differentially respond to hierarchical stimuli. ERP data indicated that women had P100 responses that were selectively lateralized to the left hemisphere in response to local targets and N150 responses that were smaller for global targets. They also had P300 responses that were greater following local stimuli. The physiological data demonstrate that male-female performance differences arise from biologically based differences in hemispheric asymmetry. Findings are discussed in the context of existing literature regarding gender differences, hemispheric specialization, and the role of stimulus characteristics. PMID- 16271818 TI - Some decks are better than others: the effect of reinforcer type and task instructions on learning in the Iowa Gambling Task. AB - The Iowa Gambling Task (Bechara, Damasio, Damasio, & Anderson, 1994) has become widely used as a laboratory test of "real-life" decision-making. However, aspects of its administration that have been varied by researchers may differentially affect performance and the conclusions researchers can draw. Some researchers have used facsimile money reinforcers while others have used real money reinforcers. More importantly, the instructions participants receive have also been varied. While no differences have been reported in performance dependent on reinforcer type, no previous comparison of participants' instructions has been conducted. This is despite one set of instructions giving participants a clear hint about the nature of the task. Additionally, in previous research one set of instructions have not been used exclusively with one reinforcer type making any differential or cumulative effects of these factors difficult to interpret. The present study compared the effects of instruction and reinforcer type on IGT performance. When participants received instructions without a hint performance was affected by reinforcer type. This was not the case when the instructions included a hint. In a second IGT session performance was improved in participants who had received the hint instructions compared with those who had not. PMID- 16271819 TI - Reading disability and hemispheric interaction on a lexical decision task. AB - The assumptions tested were that the relative contribution of each hemisphere to reading alters with experience and that experience increases suppression of the simultaneous use of identical strategies by the non-dominant hemisphere. Males that were reading disabled and phonologically impaired, reading disabled and phonologically normal, or with no reading disability were presented familiar words, orthographically correct pseudowords, and orthographically incorrect non words for lexical decision. Accuracy and response times in all groups showed a shift from no asymmetry in processing non-words to a stable left hemisphere advantage and clear suppression of the right hemisphere in processing words. In the pseudoword condition, accuracy scores were higher when both hemispheres were free to engage, especially in those with a reading disability and responses slowed in the phonologically impaired group but not the phonologically normal groups when the right hemisphere was disengaged. As familiar words typically invoke lexical processing by both hemispheres while pseudowords invoke lexical processing by the right and non-lexical processing by the left hemisphere, and as non-lexical processing is weak in the phonologically impaired, the results support the assumptions that were tested. PMID- 16271820 TI - Color makes a difference: two-dimensional object naming in literate and illiterate subjects. AB - Previous work has shown that illiterate subjects are better at naming two dimensional representations of real objects when presented as colored photos as compared to black and white drawings. This raises the question if color or textural details selectively improve object recognition and naming in illiterate compared to literate subjects. In this study, we investigated whether the surface texture and/or color of objects is used to access stored object knowledge in illiterate subjects. A group of illiterate subjects and a matched literate control group were compared on an immediate object naming task with four conditions: color and black and white (i.e., grey-scaled) photos, as well as color and black and white (i.e., grey-scaled) drawings of common everyday objects. The results show that illiterate subjects perform significantly better when the stimuli are colored and this effect is independent of the photographic detail. In addition, there were significant differences between the literacy groups in the black and white condition for both drawings and photos. These results suggest that color object information contributes to object recognition. This effect was particularly prominent in the illiterate group. PMID- 16271821 TI - Limbic system mechanisms of stress regulation: hypothalamo-pituitary adrenocortical axis. AB - Limbic dysfunction and hypothalamo-pituitary-adrenocortical (HPA) axis dysregulation are key features of affective disorders. The following review summarizes our current understanding of the relationship between limbic structures and control of ACTH and glucocorticoid release, focusing on the hippocampus, medial prefrontal cortex and amygdala. In general, the hippocampus and anterior cingulate/prelimbic cortex inhibit stress-induced HPA activation, whereas the amygdala and perhaps the infralimbic cortex may enhance glucocorticoid secretion. Several characteristics of limbic-HPA interaction are notable: first, in all cases, the role of given limbic structures is both region- and stimulus-specific. Second, limbic sites have minimal direct projections to HPA effector neurons of the paraventricular nucleus (PVN); hippocampal, cortical and amygdalar efferents apparently relay with neurons in the bed nucleus of the stria terminalis, hypothalamus and brainstem to access corticotropin releasing hormone neurons. Third, hippocampal, cortical and amygdalar projection pathways show extensive overlap in regions such as the bed nucleus of the stria terminalis, hypothalamus and perhaps brainstem, implying that limbic information may be integrated at subcortical relay sites prior to accessing the PVN. Fourth, these limbic sites also show divergent projections, with the various structures having distinct subcortical targets. Finally, all regions express both glucocorticoid and mineralocorticoid receptors, allowing for glucocorticoid modulation of limbic signaling patterns. Overall, the influence of the limbic system on the HPA axis is likely the end result of the overall patterning of responses to given stimuli and glucocorticoids, with the magnitude of the secretory response determined with respect to the relative contributions of the various structures. PMID- 16271822 TI - Inhibition of human cytochrome CYP 1 enzymes by flavonoids of St. John's wort. AB - CYP 1B1 is involved in metabolizing both polycyclic aromatic hydrocarbons and estradiol to potentially carcinogenic intermediates, and it is also over expressed in human cancer cells. In order to investigate whether flavonoids could specifically inhibit CYP 1B1, seven flavonoids in St. John's wort and apigenin were screened for their inhibition of recombinant human CYP 1B1 and CYP 1A1. While seven flavonoids (myricetin, apigenin, kaempferol, quercetin, amentoflavone, quercitrin and rutin) were slightly more selective for CYP 1B1 EROD inhibition (K(i)s 0.06-5.96 microM) compared to CYP 1A1 (K(i)s 0.20-1.6 microM) the difference in K(i)s for the P450s were not significantly different. Rutin did not inhibit CYP 1A1 at concentrations up to 10 microM. Kinetic analyses determined that apigenin and amentoflavone were competitive inhibitors of CYP 1B1, while quercetin showed mixed type inhibition. To characterize the inhibition potential of these flavonoids, five were studied further for their ability to inhibit TCDD-induced EROD activity in 22Rv1 human prostate cancer cells. 22Rv1 cells express constitutive and TCDD-inducible CYP 1A1 and CYP 1B1 mRNA. In the cells, the IC(50)s were similar to those measured for the recombinant CYP 1A1 except for amentoflavone. Quercetin (IC(50): 4.1 microM), kaempferol (3.8 microM), myricetin (3.0 microM) and apigenin (3.1 microM) caused significant inhibition of EROD activity whereas amentoflavone did not cause inhibition. Depending on their bioavailability, flavonoids that can selectively inhibit CYP1 enzymes may be useful as chemoprotective agents in prostate cancer prevention. PMID- 16271823 TI - Searching for hypothetical proteins: theory and practice based upon original data and literature. AB - A large part of mammalian proteomes is represented by hypothetical proteins (HP), i.e. proteins predicted from nucleic acid sequences only and protein sequences with unknown function. Databases are far from being complete and errors are expected. The legion of HP is awaiting experiments to show their existence at the protein level and subsequent bioinformatic handling in order to assign proteins a tentative function is mandatory. Two-dimensional gel-electrophoresis with subsequent mass spectrometrical identification of protein spots is an appropriate tool to search for HP in the high-throughput mode. Spots are identified by MS or by MS/MS measurements (MALDI-TOF, MALDI-TOF-TOF) and subsequent software as e.g. Mascot or ProFound. In many cases proteins can thus be unambiguously identified and characterised; if this is not the case, de novo sequencing or Q-TOF analysis is warranted. If the protein is not identified, the sequence is being sent to databases for BLAST searches to determine identities/similarities or homologies to known proteins. If no significant identity to known structures is observed, the protein sequence is examined for the presence of functional domains (databases PROSITE, PRINTS, InterPro, ProDom, Pfam and SMART), subjected to searches for motifs (ELM) and finally protein-protein interaction databases (InterWeaver, STRING) are consulted or predictions from conformations are performed. We here provide information about hypothetical proteins in terms of protein chemical analysis, independent of antibody availability and specificity and bioinformatic handling to contribute to the extension/completion of protein databases and include original work on HP in the brain to illustrate the processes of HP identification and functional assignment. PMID- 16271824 TI - A mathematical model to determine molecular kinetic rate constants under non steady state conditions using fluorescence recovery after photobleaching (FRAP). AB - Fluorescence recovery after photobleaching (FRAP) analyses of binding and unbinding of molecules that interact with insoluble scaffolds, such as the cytoskeleton and nuclear matrix, in living cells commonly assume that this process is at equilibrium over the time scale of fluorescence recovery. This assumption breaks down for relatively fast intracellular processes like focal adhesion assembly at the leading edge of a migrating cell, or changes of transcriptional activation in the nucleus, that can occur in a matter of a few minutes. In this paper, we formulate a mathematical model that permits FRAP to be used to determine kinetic rate constants of molecules that interact with insoluble cellular structures under non-steady state conditions. We show that unlike steady state FRAP, fluorescence recovery time scales under these unsteady conditions are determined not only by unbinding rates, but also by the overall assembly and disassembly dynamics of the structural scaffold which supports these binding interactions. Experimental data from FRAP analysis and quantification of scaffold assembly dynamics may be combined and used with our mathematical model to estimate kinetic rate constants, as well as the apparent rate constant of scaffold assembly and disassembly. PMID- 16271825 TI - Late-onset hereditary sensory neuropathy type I due to SPTLC1 mutation: autopsy findings. AB - There is little published information on the autopsy findings in hereditary sensory neuropathy type I (HSN I), and none in genetically confirmed cases. We report the neuropathological findings in a 93-year-old woman with a disease of unusually late onset, who was part of a large HSN I kindred and in whom genetic analysis confirmed an SPTLC1 T399G mutation. PMID- 16271826 TI - Capture of mercury ions by natural and industrial materials. AB - In this paper the technical feasibility of various adsorbents for mercury removal from contaminated waters has been studied. Adsorption isotherms of mercury ions in aqueous solution have been experimentally measured on a granular activated carbon (Aquacarb 207EA), a char, a pozzolana and a yellow tuff. The experimental evidences show that the mercury capture capacity of yellow tuff and char is of few tenths of milligrams per gram of sorbent while for the pozzolana and the activated carbon this value is of the order of 1mg/g of sorbent. Moreover, for a mercury concentration as high as 3000 microg/l the pozzolana shows the highest adsorption capacity. This result seems to be quite interesting, especially in consideration of the extremely low cost of this natural sorbent. PMID- 16271828 TI - Re: "Exposure of a liquefied gas container to an external fire" by Phani K. Raj, J. Hazard Mater. A122 (2005) 37-49. PMID- 16271827 TI - Mechanism of uranium removal from the aqueous solution by elemental iron. AB - The effectiveness of elemental iron (Fe(0)) to remove uranium (U) from the aqueous phase has been demonstrated. While the mitigation effect is sure, discrepancies in the removal mechanism have been reported. The objective of this study was to investigate the mechanism of U(VI) removal from aqueous phases by Fe(0). For this purpose, a systematic sequence of bulk experiments was conducted to characterize the effects of the availability and the abundance of corrosion products on U(VI) removal. Results indicated that U(VI) removal reactions did not primary occur at the surface of the metallic iron. It is determined that U(VI) co precipitation with aging corrosion products is a plausible explanation for the irreversible fixation under experimental conditions. Results of XRD analyses did no show any U phases, whereas SEM-EDX analyses showed that U tended to associate with rusted areas on the surface of Fe(0). Recovering U with different leaching solutions varied upon the dissolution capacity of the individual solutions for corrosion products, showing that the irreversibility of the removal depends on the stability of the corrosion products. U(VI) co-precipitation as removal mechanism enables a better discussion of reported discrepancies. PMID- 16271829 TI - A review of large-scale LNG spills: experiments and modeling. AB - The prediction of the possible hazards associated with the storage and transportation of liquefied natural gas (LNG) by ship has motivated a substantial number of experimental and analytical studies. This paper reviews the experimental and analytical work performed to date on large-scale spills of LNG. Specifically, experiments on the dispersion of LNG, as well as experiments of LNG fires from spills on water and land are reviewed. Explosion, pool boiling, and rapid phase transition (RPT) explosion studies are described and discussed, as well as models used to predict dispersion and thermal hazard distances. Although there have been significant advances in understanding the behavior of LNG spills, technical knowledge gaps to improve hazard prediction are identified. Some of these gaps can be addressed with current modeling and testing capabilities. A discussion of the state of knowledge and recommendations to further improve the understanding of the behavior of LNG spills on water is provided. PMID- 16271830 TI - Zeolite synthesis from paper sludge ash at low temperature (90 degrees C) with addition of diatomite. AB - Paper sludge ash was partially converted into zeolites by reaction with 3M NaOH solution at 90 degrees C for 24 h. The paper sludge ash had a low abundance of Si and significant Ca content, due to the presence of calcite that was used as a paper filler. Diatomite was added to the NaOH solution to increase its Si content in order to synthesize zeolites with high cation exchange capacity. Diatomite residue was filtered from solution before addition of ash. The original ash without addition of diatomite yielded hydroxysodalite with a cation exchange capacity ca. 50 cmol/kg. Addition of Si to the solution yielded Na-P1 (zeolite-P) with a higher cation exchange capacity (ca. 130 cmol/kg). The observed concentrations of Si and Al in the solution during the reaction explain the crystallization of these two phases. The reaction products were tested for their capacity for PO(4)(3-) removal from solution as a function of Ca(2+) content, suggesting the formation of an insoluble Ca-phosphate salt. The product with Na P1 exhibits the ability to remove NH(4)(+) as well as PO(4)(3-) from solution in concentrations sufficient for application in water purification. Both NH(4)(+) and PO(4)(3-) removal showed little variation with pH between 5 and 9. Alternative processing methods of zeolite synthesis, including the addition of ash to an unfiltered Si-NaOH solution and addition of a dry ash/diatomite mixture to NaOH solution, were tested. The third process yielded materials with lower cation exchange capacity due to formation of hydroxysodalite. The second process results in a product with relatively high cation exchange capacity, and reduces the number of processing steps necessary for zeolite synthesis. PMID- 16271831 TI - Evidence that brain nitric oxide inhibition increases metabolic cost of exercise, reducing running performance in rats. AB - To assess the role of nitric oxide (NO) in the metabolic rate and running performance of rats submitted to exercise on a treadmill, 1.43 micromol (2 microL) of Nomega-nitro-L-arginine methyl ester (L-NAME, n=6), a NO synthase inhibitor, or 2 microL of 0.15M NaCl (SAL, n=6) was injected into the lateral cerebral ventricle of male Wistar rats immediately before the animals started running (18m min(-1), 5% inclination). Oxygen consumption (VO2) was measured at rest, during the exercise until fatigue and thereafter during the 30 min of recovery using the indirect calorimetry system. Mechanical efficiency (ME) was also calculated during the running period. During the first 11 min of exercise, there was a similar increase in VO2 while ME remained the same in both groups. Thereafter, VO2 remained stable in the SAL group but continued to increase and remained higher in the L-NAME group until fatigue. The L-NAME-treated rats also showed a sharper decrease in ME than controls. In addition, there was a significant reduction in workload performance by L-NAME-treated animals compared to SAL-treated animals. This suggests that central blockage of nitric oxide increases metabolic cost during exercise, reduces mechanical efficiency and decreases running performance in rats. PMID- 16271832 TI - Relationship between toxicity of selected insecticides and expression of stress proteins (HSP, GRP) in cultured human cells: effects of commercial formulations versus pure active molecules. AB - Three carbamate (formetanate, methomyl, pyrimicarb) and one pyrethroid (bifenthrin) insecticides were investigated both as pure chemicals and as commercial formulations in order to unveil possible toxic effects of additives and solvents present in the commercial formulations and to evaluate the cellular stress response as a defense mechanism. Toxic effects were evaluated on A549 cells, derived from a human lung carcinoma, by measuring (1) threshold concentrations leading to a decrease of the growth rate (LOEC), (2) sublethal concentrations (SC) which arrested growth without killing the cells, and (3) expression levels of several stress proteins, i.e., HSP27, HSP72/73, HSP90, GRP78, and GRP94. As compared to the pure active molecule, LOEC appeared at lower concentrations when using the commercial formulations, i.e., Dicarzol (formetanate), Lannate20 (methomyl) and Talstar or Kiros EV (bifenthrin). Propylene glycol and propylene glycol monomethyl ether, respectively, present in Talstar and kiros, do not account for the high toxicity of these commercial formulations and do not potentiate the toxicity of bifenthrin. Additive but not synergistic adverse effects were observed when cells are exposed to a mixture of 4 different commercial formulations. Our results show that the concentrations of active molecules recommended in flori-cultural general use or for spray preparations are much higher than SC concentrations, as determined on A549 pulmonary cells. GRP78 was up-regulated by all the insecticides, commercial preparations being more efficient to trigger the stress reaction. This suggests that insecticides and additives present in commercial formulations disrupt ER functions. Conversely, HSP72/73 was found to be down-regulated by all the insecticides. This seems to be related with a decrease of protein synthesis in the cytosol, as a result of the ER unfolded protein response. Indeed, tunicamycin, known to inhibit N-linked glycosylation in the ER, was found to induce a similar inverse correlation between GRP78 overexpression and HSP72/73 under-expression. Expression of GRP94 was found to be increased and HSP27 lowered by the highest concentrations of bifenthrin commercial formulations. Methomyl and Lannate20 only induced an under-expression of HSP90. PMID- 16271833 TI - Increase in phosphotidylinositide-3 kinase activity by nitrotyrosylation of lysates of platelets from patients with systemic sclerosis. AB - We have observed that the platelet non-integrin collagen receptor (65 kDa) and another protein (M(r) 185 kDa) are altered in the posttranslational modification by nitrotyrosylation in platelets from patients with systemic sclerosis (SSc). We reported the identification of nitrotyrosylated 65-kDa proteins in a previous study. In the present investigation, using Western blots, one- and two dimensional gel electrophoreses and matrix assisted ionization/desorption-time of flight (MALDI-TOF) we have identified the 185-kDa protein as phosphoinositide kinase C2beta (PI 3-K). There is a positive correlation between the nitrotyrosylation of PI 3-K and activity of the enzyme, i.e., the nitrotyrosylation of PI 3-K increases its enzymatic activity. In addition, the activity of PI 3-K increases in nitrotyrosylated platelet lystaes from patients with SSc compared to normal volunteer controls, suggesting that this is an alteration in the posttranslational modification of PI 3-K in platelets from patients with SSc. The increased nitrotyrosylation of PI 3-K may contribute to the impairment of platelet function in patients with SSc by increasing platelet reactivity to matrix components within the vascular walls of patients with this disease. PMID- 16271834 TI - Serological characterization of Haemophilus parasuis isolates from China. AB - From September 2002 to December 2004, a total of 281 strains of Haemophilus parasuis were isolated from 17 provinces of China. All these isolates were serotyped by both the gel diffusion (GD) and the indirect haemagglutination (IHA) tests. By combining the GD and IHA results, serovar 4 (24.2%) and serovar 5 (19.2%) were the most prevalent serovars, followed by serovars 13 (12.5%), 14 (7.1%) and 12 (6.8%), while 12.1% of the isolates could not be assigned to a serovar (nontypable). A comparison of the number of isolates obtained from the respiratory tract of swine without polyserositis with those obtained from swine with polyserositis revealed an increased frequency of serovar 4 and a significantly decreased frequency of serovar 13 among isolates from the respiratory tract of swine without polyserositis, whereas the frequency of isolation of serovars 5, 12, 14 and nontypable from swine with or without polyserositis were similar. Co-infection of H. parasuis and other bacterial agents was studied in 183 cases examined from June 2003 to December 2004. Streptococcus suis (30.6%; 56), Escherichia coli (21.9%; 40), Bordetella bronchiseptica (21.3%; 39) and Pasteurella multocida (14.2%; 26) were the bacterial agents frequently co-isolated with H. parasuis in China. PMID- 16271835 TI - Growth hormone therapy in short children born small for gestational age. AB - Being born small for gestational age (SGA) is one of the most common causes of childhood short stature, and recombinant GH therapy has been recently licensed to promote growth in short SGA children from the age of 4 years old. Studies are now reporting very encouraging effects on adult height gains, especially in those children who started GH therapy early, at least 2 years prior to the onset of puberty. Compared to the age at starting treatment, the GH dose has a less significant impact on final height, and more attention needs to be paid now to identify earlier those SGA children who fail to catch-up spontaneously. The benefits are not just in terms of height, but also in body composition and possibly blood pressure and lipid levels. However the risk of side effects and long-term complications, particularly related to the expected metabolic effects of GH in inducing insulin resistance and hyperinsulinaemia, need to be carefully monitored especially in SGA children with a family history of type 2 diabetes. Recently, GH therapy was found to amplify the adrenarche of short SGA children and to induce a pro-inflammatory shift, as judged by a rise of neutrophil count and circulating interleukin-6 (IL-6), and a fall in adiponectin levels. Further progress is anticipated to assess the addition of insulin-sensitizing therapy to attenuate the GH-induced hyperinsulinemia, in order to alter the pro-inflammatory course, to avoid excessive release of adrenal androgens, and to slow down the potential rapid tempo of pubertal progression in SGA children. In the meantime, post-SGA short stature is rapidly becoming one of the prime indications for GH therapy in childhood. PMID- 16271836 TI - Anti-diabetic activity of methanol/methylene chloride stem bark extracts of Terminalia superba and Canarium schweinfurthii on streptozotocin-induced diabetic rats. AB - Stem bark extracts of Terminalia superba Engl. and Diels and Canarium schweinfurthii Engl. are used in Africa for the treatment of various ailments, including diabetes mellitus. The anti-diabetic effects of the methanol/methylene chloride extracts of the stem barks on streptozotocin (STZ)-induced diabetes were evaluated on male rats. Through the subcutaneous route, diabetes was induced using 60 mg/mL of streptozotocin. After 2 days, the rats received, by gavage, 150 mg/kg and 300 mg/kg of extract daily for 14 days. At 300 mg/kg, the two extracts (Terminalia superba and Canarium schweinfurthii), significantly showed at least 67.1% and 69.9% reduction in blood glucose level, respectively, while insulin (three units) given subcutaneously and once daily, had 76.8% reduction compared to diabetic untreated control rats. Similarly, the weight gains were 6.6% and 4.9%, respectively, and were comparable to the normal rats, whereas, diabetic untreated rats lost 14.1% body weight. Still with the same dose, there was 68.5% and 58.5% (p < 0.001) significant decrease in food consumption and 79.7% and 64.0% (p < 0.001) in fluid intake by diabetic rats treated with the respective plant extracts. The insulin-treated rats showed 56.4% and 75.8% decrease in food and fluid intake compared to an augmentation for diabetic control rats, 43.0% and 383.8%, respectively, at the end of the second week of experimentation. These results showed that the plant extracts can reverse hyperglycemia, polyphagia and polydipsia provoked by streptozotocin, and thus, they have anti-diabetic properties. PMID- 16271838 TI - Bayesian networks and probabilistic reasoning about scientific evidence when there is a lack of data. AB - Bayesian networks (BNs) are a kind of graphical model that formally combines elements of graph and probability theory. BNs are a mathematically and statistically rigorous technique allowing their user to define a pictorial representation of assumed dependencies and influences among a set of variables deemed to be relevant for a particular inferential problem. The formalism allows one to process newly acquired evidence according to the rules of probability calculus. Applications of BNs have been reported in various forensic disciplines. However, there seems to be some reluctance to consider BNs as a more general framework for representing and evaluating sources of uncertainties associated with scientific evidence. Notably, BNs are widely thought of as an essentially numerical method, requiring "exact" numbers with a high "accuracy". The present paper aims to draw the reader's attention to the point that the availability of hard numerical data is not a necessary requirement for using BNs in forensic science. An abstraction of quantitative BNs, known as qualitative probabilistic networks (QPNs), and sensitivity analyses are presented and their potential applications discussed. As a main difference to their quantitative counterpart, QPNs contain qualitative probabilistic relationships instead of numerical relations. Sensitivity analyses consist of varying the probabilities assigned to one or more variables and evaluating the effect on one or more other variables of interest. Both QPNs and sensitivity analyses appear to be useful concepts that permit one to work in contexts with acute lack of numerical data and where reasoning consistent with the laws of probability should nevertheless be performed. PMID- 16271837 TI - Free radical scavenging potential of Chlorophytum tuberosum Baker. AB - Chlorophytum tuberosum Baker commonly referred as 'Musli' has been widely used as a potent 'Rasayana' drug in 'Ayurveda' as a rejuvenator and tonic. Antioxidant potential of Chlorophytum tuberosum has been investigated for their ability to scavenge 1,1,diphenyl picryl hydrazyl (DPPH), nitric oxide radical along with their capacity to reduce lipid peroxidation in rat liver homogenate, chelation of ferrous ion, radical scavenging potential using chemiluminescence and their total antioxidant capacity. Sugar, starch, protein, and Vitamin C content were estimated spectrophotometrically along with the percentages of the individual amino acids by HPLC and individual sugars by using HPTLC as standardization tool. The extract has been found to possess antioxidant activity in all the models tested as evident by IC50 values being 225.31, 888.44, 809.22 and 422.97 microg/ml for scavenging of DPPH, nitric oxide, lipid peroxidation and ferry bi pyridyl complex, respectively, along with a integral anitoxidant activity of 2.986 nmol ascorbic acid/g equivalents in photochemiluminescence assay. PMID- 16271839 TI - [Erysipelas after osteoarticular prosthesis]. AB - OBJECTIVE: The occurrence of erysipelas after implantation of osteoarticular prosthesis is rarely reported in literature except when it may indicate infection of the implant. PATIENTS AND METHODS: We studied retrospectively 77 files of patients that had been hospitalized for erysipelas from January 1999 to December 2003. RESULTS: We included 3 patients (3.8%) 2 women and a man (average age: 61 years) with a history of osteoarticular prosthesis implantation on the same side as erysipelas. The period between implantation of the prosthesis and erysipelas varied from 6 months to 30 years. The 3rd patient also presented with chronic venous insufficiency and was treated for legs ulcers of venous origin. There were neither clinical nor radiological signs of prosthesis infection. The clinical schedule was typical. The initial outcome was favorable under intravenous penicillin G and local care. Antibiotic prophylaxis was recommended for all the patients, however, 2 patients relapsed. PMID- 16271840 TI - [Efficacy and safety of telithromycin in the treatment of acute exacerbation of chronic obstructive pulmonary disease]. AB - The aim of this study was to evaluate the clinical efficacy of telithromycin administered for 5 days at a dosage of 800 mg/day, in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD) presenting with at least 2 of Anthonisen's criteria including the increase of purulence. METHODOLOGY: During this multicenter (211 private lung specialists), prospective, non-comparative, open-labeled French study, 365 patients were included between April 2002 and March 2003. Clinical efficacy was assessed on D12-D19 by the rate of clinical success as defined by recovery or clinical improvement (main endpoint) according to the number of exacerbation episodes during the previous year. RESULTS: On D12-D19 clinical success rate in the per protocol global population was 88.0% and respectively 87.9% in patients with or=4 episodes in the previous year. These success rates were similar to those in the intent-to-treat population. Safety, assessed on 359 patients, was satisfactory, with mainly digestive disorders related to the treatment in 3.9% of the patients. No treatment-related serious adverse events were observed. CONCLUSION: This study, conducted among private practitioners in France according to COPD classification as defined by official recommendations, validates the results obtained in previous studies. Our results confirm the place attributed to telithromycin in the treatment of patients presenting with AECOPD without chronic respiratory failure, according to ongoing official recommendations. PMID- 16271841 TI - [Treatment interruption in HIV infected patients: clinical and biological evolution]. AB - OBJECTIVE: The authors had for aim to evaluate the clinical and biological evolution in HIV-infected patients with viraemia lower than 30,000 copies/mL having decided to interrupt their treatment. PATIENTS AND METHODS: Patients with highly active antiretroviral therapy (HAART) for more than 3 months followed by treatment interruption longer than 1 month were included in a retrospective analysis. RESULTS: Forty-six patients having stopped treatment between November 1999 and July 2003 were included. The median duration of treatment interruption was 9.5 months. During the study, no clinical event occurred for 21 patients, and at least 1 clinical event occurred for the 25 others. The median CD4(+) cell counts (CD4) before and at the end of treatment interruption were 597/mm(3) and 437/mm(3), respectively (P<0.001). The median values of viral load before and at the end of treatment interruption were <50 and 23749 copies/mL, respectively (P<0.001). Among the 26 patients having started a new HAART, pre-treatment interruption and post-new HAART median CD4 (with a median delay after HAART of 9.7 months) were 548 and 432.5/mm(3) (P=0.02). Pre-treatment interruption and post-new HAART median viral load were 131.5 and 94.5 copies/mL (NS). CONCLUSIONS: Treatment interruption must be used with caution in spite of the absence of virological impact, because CD4 cell count after new HAART is lower than CD4 preceding treatment interruption. Treatment interruption is contraindicated for patients with AIDS. Physicians must carefully follow other patients who decide on a treatment interruption. PMID- 16271842 TI - [Imported malaria at the Marseilles Hopital-Nord, France: a prospective study on 352 cases between 2001 and 2003]. AB - OBJECTIVE: The authors had for aim to study epidemiological, clinical, and parasitological characteristics, as well as regimen received, of imported malaria cases hospitalised at the North University Hospital, in Marseilles, France. DESIGN: The patients presenting with imported malaria included in this study were hospitalised in the infectious and tropical diseases unit and in the pediatrics unit at the North University Hospital, from January 1, 2001 to December 31, 2003. Variables were prospectively collected and recorded. RESULTS: 352 patients including 240 adults and 112 children were included. Most of them (67% of the adults and 92% of the children) were contaminated during a trip to the Comoros Islands. Plasmodium falciparum was the most common species identified. 97.5% of adult and 98% of child patients back from Comoros did not take any chemoprophylaxis against malaria or took inadequate regimens. Halofantrin was the most commonly used drug for children to treat uncomplicated P. falciparum malaria. In adults, atovaquone-proguanil was used as a first line drug in the absence of vomiting, and a 3-day intravenous regimen of quinine-clindamycin in case of vomiting. CONCLUSION: The specificity of imported malaria in Marseilles is the high proportion of Comorian patients who go back home periodically to visit friends and relatives. A better education of the Comorian population in Marseilles, regarding malaria risks and prophylaxis, needs to be implemented. PMID- 16271843 TI - Protective effects of hyperoside (quercetin-3-o-galactoside) to PC12 cells against cytotoxicity induced by hydrogen peroxide and tert-butyl hydroperoxide. AB - Reactive oxygen species are believed to play a very significant role in the pathogenesis of several diseases including Alzheimer's disease and Parkinson's disease. It is reported that the crucial balance between reactive oxygen species generation and antioxidant defense is regarded as a force in a wide variety of chronic diseases. In this paper, PC12 cells were used to study the antioxidative effect of hyperoside. The results indicated that hyperoside could effectively protect PC12 cells against cytotoxicity induced by hydrogen peroxide and tert butyl hydroperoxide at 160 microg/ml and 100 microg/ml, respectively. The study also showed that hyperoside was no harmful within the tested concentration range and could easily enter into the PC12 cells. With the increasing concentration of hyperoside, cytotoxicity induced by hydrogen peroxide and tert-butyl hydroperoxide was significantly attenuated and the corresponding extracellular lactate dehydrogenase levels decreased concurrently by pretreatment with hyperoside. The results were proved by flow cytometric detection of apoptotic cells. All the above results showed hyperoside could efficiently prevent the PC12 cells from shrinking and turning against apoptosis induced by hydrogen peroxide and tert-butyl hydroperoxide. PMID- 16271844 TI - Reduced quantity and hydrogen-peroxide production of vaginal lactobacilli in HIV positive women. AB - It has been suggested that vaginal lactobacilli may influence heterosexual transmission of HIV infection. The aim of this study was to compare the vaginal flora on Gram's stained and isolation rate, quantity and H2O2 production of lactobacilli between HIV positive and HIV negative women. Although, the prevalence of abnormal vaginal flora was increased in HIV infected women, there was no significant difference in isolation rate of vaginal lactobacilli between the two groups (71.87 vs. 83.33%; P>0.05). However, the results of this study showed significantly reduced quantity of lactobacilli in HIV infected women (P<0.01). In particular, the prevalence of H2O2-producing lactobacilli was lower in HIV positive as compared to HIV negative women (80 vs. 56.52%), with borderline significance (P=0.057). Taken together, our findings showed altered vaginal microflora with reduced quantity and hydrogen-peroxide production of vaginal lactobacilli in HIV positive women, but further studies are needed to assess its actual significance and potential benefit from the use of probiotic therapy. PMID- 16271845 TI - A liposomal formulation of doxorubicin, composed of hexadecylphosphocholine (HePC): physicochemical characterization and cytotoxic activity against human cancer cell lines. AB - The overall goal of this study was to prepare a novel liposomal formulation of doxorubicin, composed of hexadecylphosphocholine (HePC), as a combined formulation and to study its activity against cancer cells and peripheral blood mononuclear cells (PBMCs), in terms of efficacy and toxicity. Liposomes composed of HePC/egg phosphatidylcholine/stearylamine (HePC/EPC/SA) 10:10:0.1 (molar ratio) (1) and EPC/SA 10:0.1 (molar ratio) (2) were prepared and doxorubicin was encapsulated using the pH gradient method. Determination of lipids and doxorubicin has been achieved by high-performance thin-layer chromatography coupled with a flame-ionization detector. Prepared liposomes were characterized for their size distribution and their zeta-potential at each step of the preparation procedure. In vitro release studies have been evaluated in buffer and culture medium at 25 and 37 degrees C for 24 hours period. Liposomal formulations, free doxorubicin and HePC were tested against cancer cell lines and PBMCs, using sulforhodamine B (SRB) assay. Doxorubicin was encapsulated into the liposomes 1 and 2 at a drug to lipid molar ratio of 1.08 and 0.77, respectively, with an entrapping efficiency almost 100% in both cases. Doxorubicin was retained into liposome 1 up to 70% at 25 degrees C in TES, while up to 80% was released from 1 when liposomes were incubated at 37 degrees C either in culture medium or in the TES buffer at 24 hours. The activity of doxorubicin was retained or slightly improved when entrapped into liposomes 1 and 2, while liposomal formulation 1 encapsulating doxorubicin was found to be less toxic against normal cells (PBMCs). The combination of HePC and doxorubicin in one combined formulations justified as an improvement of the therapeutic index (TI) of doxorubicin in terms of efficacy and toxicity. PMID- 16271847 TI - Tobacco smoke induces a persistent, but recoverable state in Chlamydia pneumoniae infection of human endothelial cells. AB - We investigated the extent to which tobacco smoke could induce persistence of Chlamydia pneumoniae in human endothelial cells. Aortic and coronary artery endothelia were infected in the absence or presence of non-cytotoxic concentrations of tobacco smoke medium. Following exposure to smoke medium, chlamydial inclusions were smaller and demonstrated fewer genome copies as determined by real-time PCR. Enumeration of inclusion-forming units (IFU) established a significant smoke-mediated, dose-dependent inhibition of elementary bodies (EB). Host cell apoptosis did not contribute to the observed restriction of productive infection. Ultrastructure analysis demonstrated an arrest in chlamydial development following smoke-exposure, with a predominance of reticulate bodies (RB) observed inside inclusions. Recovery of viable IFU was achieved with removal of smoke-medium and addition of L-tryptophan. In the presence of smoke, C. pneumoniae infection demonstrated all the characteristics of persistence in human endothelia cells. This is the first time that primary human arterial endothelial cells have been shown to support chlamydial persistence. Tobacco smoke is a well-characterized risk factor for progression of atherosclerosis, but a novel means of inducing chlamydial persistence in vascular cells. Thus, smoking may additionally contribute to atherosclerotic disease by inducing a persistent chlamydial infection in arterial endothelium. PMID- 16271846 TI - Effects of CD59 on antitumoral activities of phycocyanin from Spirulina platensis. AB - The regulatory effect of phycocyanin (PC) from Spirulina platensis on cluster of differentiation 59 (CD59) gene expression of Hela cells and antitumoral mechanism of PC was investigated in this study. PC was purified by hydroxylapatite (HA) and sephacrylHR-200 gel-filtration columns chromatography. The molecular weight of PC was determined by SDS-PAGE electrophoresis. The CD59 cDNA was inserted into the eukaryotic expression plasmid pALTER-MAX, and the recombinant vector pALTER-MAX CD59 was successfully constructed. By using cationic liposome (Lipfectamine-2000) mediated transfection method, the recombinant plasmid pALTER-MAX-CD59 and the selective marker PcDNA were cotransfected into Hela cells and normal Chinese hamster ovary (CHO) cells. Stable positive cell clones were sorted out and disposed with different concentrates of PC. The expression of CD59 protein was determined by in situ hybridization, immunofluorescence and enzyme linked immunosorbent assay (ELISA). In addition, the effect of PC on the proliferation of Hela cells was determined by MTT method and the expression of Fas protein was by immunohistochemistry. Results showed that PC can promote the expression of CD59 protein in Hela cells, hold back it is reproductions of Hela cells, and moreover, a dosage effect was found between them. Namely, with the ascendance of PC concentration, the expression quantities of CD59 protein and apoptosis inducing Fas protein increased and the multiplication activity of Hela cells declined, whereas PC was of no use to CD59 and Fas protein expression, and reproduction of normal CHO cells as well. Besides an imaginable antitumoral molecular immune mechanism of PC was brought forward and discussed. PMID- 16271849 TI - Mutagenic and cytotoxic effect of planifolin: a naphthopyranone dimer isolated from Paepalanthus planifolius. AB - A naphthopyranone dimer, named planifolin, was isolated from a methylene chloride extract of the capitula of Paepalanthus planifolius Koern. The molecule (C(31)H(26)O(10)) appeared to be made up of two monomeric portions, semi vioxanthin and paepalantine (an isocoumarin), linked by an ether bond, and it may possess several kinds of biological activity that can be related to its polyphenolic structure. Short-term tests that detect genetic damage can afford the information needed to evaluate carcinogenic risks of chemicals to humans. The Ames test, recommended for testing the mutagenicity of chemical compounds with potential pharmacological application, was used in the present study. The mutagenic activity was evaluated in Salmonella typhimurium strains TA100, TA98, TA102 and TA97a and the cytotoxic effect in McCoy cells. The in vitro cytotoxicity of planifolin to McCoy cells, tested in microculture with neutral red, showed a significant cytotoxic index (CI(50)) of 12.83 microg/mL. Planifolin showed mutagenic activity for TA100, TA98 and TA97a. The results indicate that this new naphthopyranone dimer causes mutations by substitution and by addition and deletion of bases in the sequence of DNA. Moreover, its mutagenic potential was increased by metabolic activation. PMID- 16271848 TI - Effect of capsazepine on cytosolic Ca(2+) levels and proliferation of human prostate cancer cells. AB - Capsazepine has been widely used as a selective antagonist of vanilloid type 1 receptors; however, its other in vitro effect on most cell types is unknown. In human PC3 prostate cancer cells, the effect of capsazepine on intracellular Ca(2+) concentrations ([Ca(2+)](i)) and cytotoxicity was investigated by using fura-2 and tetrazolium, respectively. Capsazepine caused a rapid rise in [Ca(2+)](i) in a concentration-dependent manner with an EC(50) value of 75 microM. Capsazepine-induced [Ca(2+)](i) rise was reduced by 60% by removal of extracellular Ca(2+), suggesting that the capsazepine-induced [Ca(2+)](i) rise was contributed by extracellular Ca(2+) influx and intracellular Ca(2+). Consistently, the capsazepine (200 microM)-induced [Ca(2+)](i) rise was decreased by La(3+) by half. In Ca(2+)-free medium, thapsigargin, an inhibitor of the endoplasmic reticulum Ca(2+)-ATPase, caused a monophasic [Ca(2+)](i) rise, after which the effect of capsazepine on [Ca(2+)](i) was inhibited by 80%. Conversely, pretreatment with capsazepine partly reduced thapsigargin-induced [Ca(2+)](i) rise. U73122, an inhibitor of phospholipase C, abolished histamine (an inositol 1,4,5-trisphosphate-dependent Ca(2+) mobilizer)-induced, but not capsazepine induced, [Ca(2+)](i) rise. These findings suggest that in human PC3 prostate cancer cells, capsazepine increases [Ca(2+)](i) by evoking Ca(2+) influx and releasing Ca(2+) from the endoplasmic reticulum via a phospholiase C-independent manner. Overnight incubation with capsazepine (200 microM) killed 37% of cells, which could not be prevented by chelating intracellular Ca(2+) with BAPTA. PMID- 16271850 TI - Sensitive apoptosis induced by microcystins in the crucian carp (Carassius auratus) lymphocytes in vitro. AB - Microcystins including leucine-arginine l-amino acid (MCLR) and arginine-arginine l-amino acid (MCRR) can inhibit several serine/threonine protein phosphatases. In this study, we focused on the efficient biomarker for analyzing toxic cyanobacteria blooms using in vitro apoptosis bioassay. We explored the existence of sensitive apoptosis induced by MCLR and MCRR on isolated lymphocytes of the crucian carp (Carassius auratus) at a low exposure level. Apoptosis was detected in vitro and was clearly distinguished by condensation of nuclear chromatin and formation of apoptotic bodies, after 2 h exposure at 1, 5, 10 nM MCLR and MCRR, respectively. Agarose gel electrophoresis further revealed DNA fragmentation (DNA ladder) caused by apoptosis. We found that MCLR and MCRR can induce lymphocyte apoptosis in a dose- and time-dependent manner with flow cytometry analysis. Our study provides the first evidence that microcystins can induce fish lymphocytes apoptosis and may impair fish immune function. PMID- 16271851 TI - Single nucleotide polymorphisms in the apoptosis receptor gene TNFRSF6. AB - The homotrimeric Fas receptor, an inducer of lymphocyte apoptosis, plays a critical role in cellular pathways of immune homeostasis and immunologic tolerance. Inherited and acquired defects in the Fas gene, TNFRSF6 (tumor necrosis factor receptor superfamily member 6) have been associated with human autoimmune lymphoproliferative syndrome (ALPS) and a spectrum of other complex autoimmune diseases and malignancies. In addition to over 60 deleterious mutations associated with dominant inhibitory defects or null mutations of TNFRSF6, several sequence variants have been noted. To facilitate interpretation of genotypes of this important locus, we sequenced DNA from unrelated, healthy Caucasians and African Americans. Two new and 12 previously recorded SNPs were confirmed, and their allele frequencies were determined. We also investigated haplotype frequencies and linkage disequilibrium (LD) coefficients for these SNPs in Caucasians. Four TNFRSF6 SNP pairs were found to be in strong LD. The TNRFSF6 SNPs are useful for linkage and loss of heterozygosity studies probing the role of Fas-mediated apoptosis in autoimmune diseases and malignancies. PMID- 16271853 TI - Star-nosed moles. PMID- 16271852 TI - Increased responsiveness to MDMA in adult rats treated neonatally with MDMA. AB - MDMA [(+/-)3,4-methylenedioxymethamphetamine, also known as ecstasy] is a popular recreational drug among women of reproductive age. The objective of this study was to investigate the long-term neurobehavioral consequences of early developmental MDMA exposure. On postnatal days (PD) 1-4, Sprague-Dawley rats received two 10 mg/kg injections of MDMA with an inter-dose interval of 4 h. Male subjects were tested in adulthood for their performance in an object-recognition memory task and for their thermal and behavioral responses to an acute MDMA challenge (10 mg/kg i.p.). Neonatal MDMA administration did not alter working memory in the object-recognition test in young adulthood (PD 68-73) and there were no differences in radiolabeled citalopram binding to the serotonin transporter at this age. However, the pretreated animals showed increased thermal dysregulation and serotonin syndrome responses (particularly headweaving stereotypy) following the MDMA challenge. These results add to the growing literature demonstrating that developmental MDMA administration can lead to long lasting functional abnormalities, and they further suggest that the offspring of ecstasy-using women may be at risk for enhanced sensitivity to this drug due to their earlier exposure. PMID- 16271854 TI - A way of selectively degrading colour constancy demonstrates the experience dependence of colour vision. PMID- 16271855 TI - Evolution of mate discrimination in a fish. PMID- 16271856 TI - Social learning: public information in insects. AB - Although it has received less coverage than in vertebrates, the study of insect social learning has a rich history with spectacular examples of how individuals extract knowledge from other animals. Several new studies on crickets and social bees have now shown how insects can adjust their behaviour adaptively by making use of cues generated inadvertently by other individuals. PMID- 16271857 TI - Cytokinesis: Rho marks the spot. AB - During cytokinesis of a eukaryotic cell, following the chromosome movements of anaphase, a contractile ring forms in the cortex midway between the segregating chromosomes and divides the cell into two daughters. Recent studies have provided new insights into the mechanism by which the site of contractile ring assembly is specified. PMID- 16271858 TI - Decision making: the virtue of patience in primates. AB - Marmoset monkeys devalue rewards requiring travel to acquire, but tamarin monkeys do not, despite the greater patience of marmosets when rewards are delayed in time. Such preference reversals, not predicted by standard economic theory, may reflect behavioral mechanisms adaptively specialized for different spatial and temporal patterns of foraging. PMID- 16271860 TI - Movement control: dedicated or distributed? AB - Neural networks in which 'commands' spread 'horizontally' across multiple neurons have been believed to mediate motor pattern variation. A recent study shows that dedicated 'vertical' neural pathways can also underlie these variations. PMID- 16271859 TI - Synaptic physiology: illuminating the road ahead. AB - A number of genetically encoded reporters of neuronal activity are being developed to assay synaptic activity with single synapse resolution. A recently engineered probe allows imaging of glutamatergic transmission with high sensitivity, and similar probes may help pave the way for optical imaging of excitatory synaptic function in vivo. PMID- 16271861 TI - Centrosome maturation: Aurora lights the way to the poles. AB - The centrosome is the main microtubule organising centre in the cell. During mitosis, centrosomes dramatically increase microtubule nucleating activity, enabling them to form a mitotic spindle. Recent studies show that Aurora A kinase promotes microtubule assembly from centrosomes through the phosphorylation of the conserved centrosomal protein TACC. PMID- 16271862 TI - Infant learning: music and the baby brain. AB - When it comes to listening to music, infants literally have a more open mind than their parents. Studies which investigate listening behaviour of babies and adults have shown that, as we learn to discriminate the musical sounds in our own environment, we become less sensitive to those of other cultures. PMID- 16271863 TI - Language evolution: what do chimpanzees have to say? AB - Although unique in important ways, language shares some properties with other animal communication systems. Comparative analyses of nonhuman primate vocalizations can shed light on the evolution of language's special features. PMID- 16271864 TI - Dorsoventral axis formation in the Drosophila embryo--shaping and transducing a morphogen gradient. AB - The graded nuclear location of the transcription factor Dorsal along the dorsoventral axis of the early Drosophila embryo provides positional information for the determination of different cell fates. Nuclear uptake of Dorsal depends on a complex signalling pathway comprising two parts: an extracellular proteolytic cascade transmits the dorsoventral polarity of the egg chamber to the early embryo and generates a gradient of active Spatzle protein, the ligand of the receptor Toll; an intracellular cascade downstream of Toll relays this graded signal to embryonic nuclei. The slope of the Dorsal gradient is not determined by diffusion of extracellular or intracellular components from a local source, but results from self-organised patterning, in which positive and negative feedback is essential to create and maintain the ratio of key factors at different levels, thereby establishing and stabilising the graded spatial information for Dorsal nuclear uptake. PMID- 16271865 TI - Bicoid determines sharp and precise target gene expression in the Drosophila embryo. AB - BACKGROUND: The activity of the Bicoid (Bcd) transcription factor is a useful example of how quantitative information contained in a smooth morphogen gradient is transformed into discrete and precise patterns of target gene expression. There are two distinct and important aspects to this process: the "sharpening" of the posterior borders of the expression domains and the "precision" of where the target genes are expressed along the length of the embryo as the syncytial embryo begins to cellularize. Although the sharpening phenomenon was observed over a decade ago, it is still poorly understood. RESULTS: Here, we show that a Bcd reporter gene containing binding sites only for Bcd is expressed, like natural targets of Bcd, in a precise domain with a sharp boundary. Analysis of embryos expressing deleted forms of Bcd indicates that the sharpness of the Bcd target gene hunchback's expression involves the glutamine-rich and C-terminal activation domains of Bcd. Furthermore, several artificial Gal4-derived transcription factors expressed as gradients in the embryo share Bcd's ability to drive precise target gene expression with sharp boundaries. CONCLUSION: Thus, contrary to recent reports proposing that the Bcd gradient is not sufficient to establish precise positional information, we show that Bcd drives precise and sharp expression of its target genes through a process that depends exclusively on its ability to activate transcription. PMID- 16271866 TI - Patterns of auxin transport and gene expression during primordium development revealed by live imaging of the Arabidopsis inflorescence meristem. AB - BACKGROUND: Plants produce leaf and flower primordia from a specialized tissue called the shoot apical meristem (SAM). Genetic studies have identified a large number of genes that affect various aspects of primordium development including positioning, growth, and differentiation. So far, however, a detailed understanding of the spatio-temporal sequence of events leading to primordium development has not been established. RESULTS: We use confocal imaging of green fluorescent protein (GFP) reporter genes in living plants to monitor the expression patterns of multiple proteins and genes involved in flower primordial developmental processes. By monitoring the expression and polarity of PINFORMED1 (PIN1), the auxin efflux facilitator, and the expression of the auxin-responsive reporter DR5, we reveal stereotypical PIN1 polarity changes which, together with auxin induction experiments, suggest that cycles of auxin build-up and depletion accompany, and may direct, different stages of primordium development. Imaging of multiple GFP-protein fusions shows that these dynamics also correlate with the specification of primordial boundary domains, organ polarity axes, and the sites of floral meristem initiation. CONCLUSIONS: These results provide new insight into auxin transport dynamics during primordial positioning and suggest a role for auxin transport in influencing primordial cell type. PMID- 16271867 TI - Mutations in a conserved replication protein suppress transcriptional gene silencing in a DNA-methylation-independent manner in Arabidopsis. AB - Mutations in the DNA glycosylase/lyase ROS1 cause transcriptional silencing of the linked RD29A-LUC and 35S-NPTII transgenes in Arabidopsis. We report here that mutations in the Arabidopsis RPA2 locus release the silencing of 35S-NPTII but not RD29A-LUC in the ros1 mutant background. The rpa2 mutation also leads to enhanced expression of some transposons. Neither DNA methylation nor siRNAs at any of the reactivated loci are blocked by rpa2. Histone H3 methylation at lysine 4 was increased and histone H3 methylation at lysine 9 was decreased at the 35S promoter in the ros1rpa2 mutant compared to the ros1 background. RPA2 encodes a nuclear protein similar to the second subunit of the replication protein A conserved from yeast to mammals. Ectopic expression of the Arabidopsis RPA2 could complement the yeast rfa2 (rpa2) mutant. These results suggest an essential role of RPA2 in the maintenance of transcriptional gene silencing at specific loci in a DNA-methylation-independent manner. In addition, we found that rpa2 mutants are hypersensitive to the genotoxic agent methyl methanesulphonate, and the RPA2 protein interacts with ROS1 in vitro and in vivo, suggesting that RPA2 also functions together with ROS1 in DNA repair. PMID- 16271868 TI - Arabidopsis RPA2: a genetic link among transcriptional gene silencing, DNA repair, and DNA replication. AB - Transcriptional gene silencing (TGS) controls the expression of transposable elements and of endogenous genes containing promoter repeats, and it is associated with increased DNA methylation. TGS-deficient mutants impaired in siRNA accumulation and/or chromatin modification (ago4, bru1, cmt3, dcl3, ddm1, drd1, drm2, fas1, fas2, hda6, hog1, met1, mom1, nrpd1a, nrpd1b, nrpd2a, rdr2, suvh2, and suvh4) have been identified, but not all mutations affect the same subset of targets. Here, we identify Arabidopsis RPA2, a conserved protein with DNA replication and DNA repair motifs, as a novel TGS component that is dispensable for endogenous small RNA accumulation. bru1, cmt3, ddm1, fas1, fas2, hda6, hog1, met1, mom1, and rpa2 mutants are impaired in TGS of dispersed Athila/TSI retrotransposons and of the transgene repeat locus L5, but unlike bru1, cmt3, ddm1, fas1, fas2, hda6, hog1, and met1, the rpa2 and mom1 mutants do not affect the accumulation of 5S-derived siRNAs. Like BRU1, FAS1, FAS2, and MOM1, RPA2 is dispensable for DNA methylation, and rpa2, bru1, fas1, and fas2, but not mom1, mutants are hypersensitive to the DNA damage agent MMS. These results suggest a coordination of the TGS machinery with DNA replication, repair, or recombination machinery at some loci, and they emphasize the diversification of the TGS pathway. PMID- 16271869 TI - Acehnese reefs in the wake of the Asian tsunami. AB - The Sumatra-Andanaman tsunami was one of the greatest natural disasters in recorded human history. Here, we show that on the northwest coast of Aceh, Indonesia, where the tsunami was most ferocious, the damage to corals, although occasionally spectacular, was surprisingly limited. We detected no change in shallow coral assemblages between March 2003 and March 2005, with the exception of one site smothered by sediment. Direct tsunami damage was dependent on habitat and largely restricted to corals growing in unconsolidated substrata, a feature unique to tsunami disturbance. Reef condition, however, varied widely within the region and was clearly correlated with human impacts prior to the tsunami. Where fishing has been controlled, coral cover was high. In contrast, reefs exposed to destructive fishing had low coral cover and high algal cover, a phase shift the tsunami may exacerbate with an influx of sediments and nutrients. Healthy reefs did not mitigate the damage on land. Inundation distance was largely determined by wave height and coastal topography. We conclude that although chronic human misuse has been much more destructive to reefs in Aceh than this rare natural disturbance, human modification of the reef did not contribute to the magnitude of damage on land. PMID- 16271870 TI - Social learning in noncolonial insects? AB - Social-information use has generated great interest lately and has been shown to have important implications for the ecology and evolution of species. Learning about predators or predation risk from others may provide low-cost life-saving information and would be expected to have adaptive payoffs in any species where conspecifics are observable and behave differently under predation risk. Yet, social learning and social-information use in general have been largely restricted to vertebrates. Here, we show that crickets adapt their predator avoidance behavior after having observed the behavior of knowledgeable others and maintain these behavioral changes lastingly after demonstrators are gone. These results point toward social learning, a contingency never shown before in noncolonial insects. We show that these long-lasting changes cannot instead be attributed to long re-emergence times, long-lasting effects of alarm pheromones, or residual odor cues. Our findings imply that social learning is likely much more phylogenetically widespread than currently acknowledged and that reliance on social information is determined by ecological rather than taxonomic constrains, and they question the generally held assumption that social learning is restricted to large-brained animals assumed to possess superior cognitive abilities. PMID- 16271871 TI - MOS2, a protein containing G-patch and KOW motifs, is essential for innate immunity in Arabidopsis thaliana. AB - Innate immunity is critical for sensing and defending against microbial infections in multicellular organisms. In plants, disease resistance genes (R genes) play central roles in recognizing pathogens and initiating downstream defense cascades. Arabidopsis SNC1 encodes a TIR-NBS-LRR-type R protein with a similar structure to nucleotide binding oligomerization domain (Nod) proteins in animals. A point mutation in the region between the NBS and LRR of SNC1 results in constitutive activation of defense responses in the snc1 mutant. Here, we report the identification and characterization of mos2-1, a mutant suppressing the constitutive defense responses in snc1. Analysis of mos2 single mutants indicated that it is not only required for resistance specified by multiple R genes, but also for basal resistance. Map-based cloning of MOS2 revealed that it encodes a novel nuclear protein that contains one G-patch and two KOW domains and has homologs across the animal kingdom. The presence of both G-patch and KOW domains in the MOS2 protein suggests that it probably functions as an RNA binding protein critical for plant innate immunity. Our discovery on the biological functions of MOS2 will shed light on functions of the MOS2 homologs in animals, where they may also play important roles in innate immunity. PMID- 16271872 TI - Mechanisms for allocating auditory attention: an auditory saliency map. AB - Our nervous system is confronted with a barrage of sensory stimuli, but neural resources are limited and not all stimuli can be processed to the same extent. Mechanisms exist to bias attention toward the particularly salient events, thereby providing a weighted representation of our environment. Our understanding of these mechanisms is still limited, but theoretical models can replicate such a weighting of sensory inputs and provide a basis for understanding the underlying principles. Here, we describe such a model for the auditory system-an auditory saliency map. We experimentally validate the model on natural acoustical scenarios, demonstrating that it reproduces human judgments of auditory saliency and predicts the detectability of salient sounds embedded in noisy backgrounds. In addition, it also predicts the natural orienting behavior of naive macaque monkeys to the same salient stimuli. The structure of the suggested model is identical to that of successfully used visual saliency maps. Hence, we conclude that saliency is determined either by implementing similar mechanisms in different unisensory pathways or by the same mechanism in multisensory areas. In any case, our results demonstrate that different primate sensory systems rely on common principles for extracting relevant sensory events. PMID- 16271873 TI - Distinct contributions of T1R2 and T1R3 taste receptor subunits to the detection of sweet stimuli. AB - Animals utilize hundreds of distinct G protein-coupled receptor (GPCR)-type chemosensory receptors to detect a diverse array of chemical signals in their environment, including odors, pheromones, and tastants. However, the molecular mechanisms by which these receptors selectively interact with their cognate ligands remain poorly understood. There is growing evidence that many chemosensory receptors exist in multimeric complexes, though little is known about the relative contributions of individual subunits to receptor functions. Here, we report that each of the two subunits in the heteromeric T1R2:T1R3 sweet taste receptor binds sweet stimuli though with distinct affinities and conformational changes. Furthermore, ligand affinities for T1R3 are drastically reduced by the introduction of a single amino acid change associated with decreased sweet taste sensitivity in behaving mice. Thus, individual T1R subunits increase the receptive range of the sweet taste receptor, offering a functional mechanism for phenotypic variations in sweet taste. PMID- 16271875 TI - The HMG-CoA reductase inhibitor lovastatin reverses the learning and attention deficits in a mouse model of neurofibromatosis type 1. AB - Neurofibromatosis Type 1 (NF1) is a common neurological disorder caused by mutations in the gene encoding Neurofibromin, a p21Ras GTPase Activating Protein (GAP). Importantly, NF1 causes learning disabilities and attention deficits. A previous study showed that the learning and memory deficits of a mouse model of NF1 (nf1+/-) appear to be caused by excessive p21Ras activity leading to impairments in long-term potentiation (LTP), a cellular mechanism of learning and memory. Here, we identify lovastatin as a potent inhibitor of p21Ras/Mitogen Activated Protein Kinase (MAPK) activity in the brain. Lovastatin is a specific inhibitor of three-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, used commonly for the treatment of hypercholesterolemia. We report that lovastatin decreased the enhanced brain p21Ras-MAPK activity of the nf1+/- mice, rescued their LTP deficits, and reversed their spatial learning and attention impairments. Therefore, these results demonstrate that lovastatin may prove useful in the treatment of Neurofibromatosis Type 1. PMID- 16271874 TI - Punishment prediction by dopaminergic neurons in Drosophila. AB - The temporal pairing of a neutral stimulus with a reinforcer (reward or punishment) can lead to classical conditioning, a simple form of learning in which the animal assigns a value (positive or negative) to the formerly neutral stimulus. Olfactory classical conditioning in Drosophila is a prime model for the analysis of the molecular and neuronal substrate of this type of learning and memory. Neuronal correlates of associative plasticity have been identified in several regions of the insect brain. In particular, the mushroom bodies have been shown to be necessary for aversive olfactory memory formation. However, little is known about which neurons mediate the reinforcing stimulus. Using functional optical imaging, we now show that dopaminergic projections to the mushroom-body lobes are weakly activated by odor stimuli but respond strongly to electric shocks. However, after one of two odors is paired several times with an electric shock, odor-evoked activity is significantly prolonged only for the "punished" odor. Whereas dopaminergic neurons mediate rewarding reinforcement in mammals, our data suggest a role for aversive reinforcement in Drosophila. However, the dopaminergic neurons' capability of mediating and predicting a reinforcing stimulus appears to be conserved between Drosophila and mammals. PMID- 16271876 TI - Hypermutability impedes cooperation in pathogenic bacteria. AB - When the supply of beneficial mutations limits adaptation, bacterial mutator alleles can reach high frequencies by hitchhiking with advantageous mutations. However, when populations are well adapted to their environments, the increased rate of deleterious mutations makes hypermutability selectively disadvantageous. Here, we consider a further cost of hypermutability: its potential to break down cooperation (group-beneficial behavior that is costly to the individual). This probably occurs for three reasons. First, an increased rate at which 'cheating' genotypes are generated; second, an increased probability of producing efficient cheats; and third, a decrease in relatedness (not addressed in the present study). We used Pseudomonas aeruginosa's production of extracellular iron scavenging molecules, siderophores, to determine if cheating evolved more readily in mutator populations. Siderophore production is costly to individual bacteria but benefits all nearby cells. Siderophore-deficient cheats therefore have a selective advantage within populations. We observed the de novo evolution and subsequent increase in frequency of siderophore cheats within both wild-type and mutator populations for 200 generations. Cheats appeared and increased in frequency more rapidly in mutator populations. The presence of cheats was costly to the group, as shown by a negative correlation between cheat frequency and population density. PMID- 16271877 TI - EGO-1, a putative RNA-dependent RNA polymerase, is required for heterochromatin assembly on unpaired dna during C. elegans meiosis. AB - During meiosis in C. elegans, unpaired chromosomes and chromosomal regions accumulate high levels of histone H3 lysine 9 dimethylation (H3K9me2), a modification associated with facultative heterochromatin assembly and the resulting transcriptional silencing. Meiotic silencing of unpaired DNA may be a widely conserved genome defense mechanism. The mechanisms of meiotic silencing remain unclear, although both transcriptional and posttranscriptional processes are implicated. Cellular RNA-dependent RNA polymerases (RdRPs) function in development and RNA-mediated silencing in many species and in heterochromatin assembly in S. pombe. There are four C. elegans RdRPs, including two with known germline functions. EGO-1 is required for fertility and robust germline RNAi. RRF 3 acts genetically to repress RNAi and is required for normal meiosis and spermatogenesis at elevated temperatures (S. L'Hernault, personal communication). Among C. elegans RdRPs, we find that only EGO-1 is required for H3K9me2 enrichment on unpaired chromosomal regions during meiosis. This H3K9me2 enrichment does not require Dicer or Drosha nuclease or any of several other proteins required for RNAi. ego-1 interacts genetically with him-17, another regulator of chromatin and meiosis, to promote germline development. We conclude that EGO-1 is an essential component of meiotic silencing in C. elegans. PMID- 16271878 TI - A new niche for Notch on Deltex? PMID- 16271879 TI - Syn-full behavior by T7 DNA polymerase. PMID- 16271880 TI - SARS proteomics reveals viral secrets. PMID- 16271881 TI - The ins and outs of protein synthesis. PMID- 16271882 TI - Folding of small helical proteins assisted by small-angle X-ray scattering profiles. AB - This paper reports a computational method for folding small helical proteins. The goal was to determine the overall topology of proteins given secondary structure assignment on sequence. In doing so, a Monte Carlo protocol, which combines coarse-grained normal modes and a Hamiltonian at a different scale, was developed to enhance sampling. In addition to the knowledge-based potential functions, a small-angle X-ray scattering (SAXS) profile was also used as a weak constraint for guiding the folding. The algorithm can deliver structural models with overall correct topology, which makes them similar to those of 5 approximately 6 A cryo EM density maps. The success could contribute to make the SAXS technique a fast and inexpensive solution-phase experimental method for determining the overall topology of small, soluble, but noncrystallizable, helical proteins. PMID- 16271883 TI - Structure and Notch receptor binding of the tandem WWE domain of Deltex. AB - Deltex is a cytosolic effector of Notch signaling thought to bind through its N terminal domain to the Notch receptor. Here we report the structure of the Drosophila Deltex N-terminal domain, which contains two tandem WWE sequence repeats. The WWE repeats, which adopt a novel fold, are related by an approximate two-fold axis of rotation. Although the WWE repeats are structurally distinct, they interact extensively and form a deep cleft at their junction that appears well suited for ligand binding. The two repeats are thermodynamically coupled; this coupling is mediated in part by a conserved segment that is immediately C terminal to the second WWE domain. We demonstrate that although the Deltex WWE tandem is monomeric in solution, it forms a heterodimer with the ankyrin domain of the Notch receptor. These results provide structural and functional insight into how Deltex modulates Notch signaling, and how WWE modules recognize targets for ubiquitination. PMID- 16271884 TI - The molecular mechanism for receptor-stimulated iron release from the plasma iron transport protein transferrin. AB - Human transferrin receptor 1 (TfR) binds iron-loaded transferrin (Fe-Tf) and transports it to acidic endosomes where iron is released in a TfR-facilitated process. Consistent with our hypothesis that TfR binding stimulates iron release from Fe-Tf at acidic pH by stabilizing the apo-Tf conformation, a TfR mutant (W641A/F760A-TfR) that binds Fe-Tf, but not apo-Tf, cannot stimulate iron release from Fe-Tf, and less iron is released from Fe-Tf inside cells expressing W641A/F760A-TfR than cells expressing wild-type TfR (wtTfR). Electron paramagnetic resonance spectroscopy shows that binding at acidic pH to wtTfR, but not W641A/F760A-TfR, changes the Tf iron binding site > or =30 A from the TfR W641/F760 patch. Mutation of Tf histidine residues predicted to interact with the W641/F760 patch eliminates TfR-dependent acceleration of iron release. Identification of TfR and Tf residues critical for TfR-facilitated iron release, yet distant from a Tf iron binding site, demonstrates that TfR transmits long range conformational changes and stabilizes the conformation of apo-Tf to accelerate iron release from Fe-Tf. PMID- 16271885 TI - Mycobacterium tuberculosis protein tyrosine phosphatase PtpB structure reveals a diverged fold and a buried active site. AB - Intracellular pathogenic bacteria manipulate host signal transduction pathways to facilitate infection. Mycobacterium tuberculosis protein tyrosine phosphatases (PTPs) PtpA and PtpB are thought to be secreted into host cells and interfere with unidentified signals. To illuminate the mechanisms of regulation and substrate recognition, we determined the 1.7 A resolution crystal structure of PtpB in complex with the product phosphate. The protein adopts a simplified PTP fold, which combines features of the conventional PTPs and dual-specificity phosphatases. PtpB shows two unusual elaborations--a disordered, acidic loop and a flexible, two-helix lid that covers the active site--that are specific to mycobacterial orthologs. Biochemical studies suggest that substrate mimicry in the lid may protect the phosphatase from oxidative inactivation. The insertion and deletion of large structural elements in PtpB suggest that, outside the active site module, the PTP family is under unusual selective pressure that promotes changes in overall structure. PMID- 16271886 TI - Activation process of [NiFe] hydrogenase elucidated by high-resolution X-ray analyses: conversion of the ready to the unready state. AB - Hydrogenases catalyze oxidoreduction of molecular hydrogen and have potential applications for utilizing dihydrogen as an energy source. [NiFe] hydrogenase has two different oxidized states, Ni-A (unready, exhibits a lag phase in reductive activation) and Ni-B (ready). We have succeeded in converting Ni-B to Ni-A with the use of Na2S and O2 and determining the high-resolution crystal structures of both states. Ni-B possesses a monatomic nonprotein bridging ligand at the Ni-Fe active site, whereas Ni-A has a diatomic species. The terminal atom of the bridging species of Ni-A occupies a similar position as C of the exogenous CO in the CO complex (inhibited state). The common features of the enzyme structures at the unready (Ni-A) and inhibited (CO complex) states are proposed. These findings provide useful information on the design of new systems of biomimetic dihydrogen production and fuel cell devices. PMID- 16271887 TI - Crystal structure of a complex between protein tyrosine phosphatase 1B and the insulin receptor tyrosine kinase. AB - Protein tyrosine phosphatase 1B (PTP1B) is a highly specific negative regulator of insulin receptor signaling in vivo. The determinants of PTP1B specificity for the insulin receptor versus other receptor tyrosine kinases are largely unknown. Here, we report a crystal structure at 2.3 A resolution of the catalytic domain of PTP1B (trapping mutant) in complex with the phosphorylated tyrosine kinase domain of the insulin receptor (IRK). The crystallographic asymmetric unit contains two PTP1B-IRK complexes that interact through an IRK dimer interface. Rather than binding to a phosphotyrosine in the IRK activation loop, PTP1B binds instead to the opposite side of the kinase domain, with the phosphorylated activation loops sequestered within the IRK dimer. The crystal structure provides evidence for a noncatalytic mode of interaction between PTP1B and IRK, which could be important for the selective recruitment of PTP1B to the insulin receptor. PMID- 16271888 TI - A lysine residue in the fingers subdomain of T7 DNA polymerase modulates the miscoding potential of 8-oxo-7,8-dihydroguanosine. AB - 8-oxo-7,8-dihydroguanosine (8oG) is a highly mutagenic DNA lesion that stably pairs with adenosine, forming 8oG(syn).dA(anti) Hoogsteen base pairs. DNA polymerases show different propensities to insert dCMP or dAMP opposite 8oG, but the molecular mechanisms that determine faithful or mutagenic bypass are poorly understood. Here, we report kinetic and structural data providing evidence that, in T7 DNA polymerase, residue Lys536 is responsible for attenuating the miscoding potential of 8oG. The Lys536Ala polymerase shows a significant increase in mutagenic 8oG bypass versus wild-type polymerase, and a crystal structure of the Lys536Ala mutant reveals a closed complex with an 8oG(syn).dATP mismatch in the polymerase active site, in contrast to the unproductive, open complex previously obtained by using wild-type polymerase. We propose that Lys536 acts as a steric and/or electrostatic filter that attenuates the miscoding potential of 8oG by normally interfering with the binding of 8oG in a syn conformation that pairs with dATP. PMID- 16271889 TI - Bovine mitochondrial peroxiredoxin III forms a two-ring catenane. AB - A crystal structure is reported for the C168S mutant of a typical 2-Cys peroxiredoxin III (Prx III) from bovine mitochondria at a resolution of 3.3 A. Prx III is present as a two-ring catenane comprising two interlocking dodecameric toroids that are assembled from basic dimeric units. Each ring has an external diameter of 150 A and encompasses a central cavity that is 70 A in width. The concatenated dodecamers are inclined at an angle of 55 degrees, which provides a large contact surface between the rings. Dimer-dimer contacts involved in toroid formation are hydrophobic in nature, whereas the 12 areas of contact between interlocked rings arise from polar interactions. These two major modes of subunit interaction provide important insights into possible mechanisms of catenane formation. PMID- 16271890 TI - Structural basis of severe acute respiratory syndrome coronavirus ADP-ribose-1'' phosphate dephosphorylation by a conserved domain of nsP3. AB - The crystal structure of a conserved domain of nonstructural protein 3 (nsP3) from severe acute respiratory syndrome coronavirus (SARS-CoV) has been solved by single-wavelength anomalous dispersion to 1.4 A resolution. The structure of this "X" domain, seen in many single-stranded RNA viruses, reveals a three-layered alpha/beta/alpha core with a macro-H2A-like fold. The putative active site is a solvent-exposed cleft that is conserved in its three structural homologs, yeast Ymx7, Archeoglobus fulgidus AF1521, and Er58 from E. coli. Its sequence is similar to yeast YBR022W (also known as Poa1P), a known phosphatase that acts on ADP-ribose-1''-phosphate (Appr-1''-p). The SARS nsP3 domain readily removes the 1'' phosphate group from Appr-1''-p in in vitro assays, confirming its phosphatase activity. Sequence and structure comparison of all known macro-H2A domains combined with available functional data suggests that proteins of this superfamily form an emerging group of nucleotide phosphatases that dephosphorylate Appr-1''-p. PMID- 16271891 TI - An experimental investigation of conformational fluctuations in proteins G and L. AB - The B1 domains of streptococcal proteins G and L are structurally similar, but they have different sequences and they fold differently. We have measured their NMR spectra at variable temperature using a range of concentrations of denaturant. Many residues have curved amide proton temperature dependence, indicating that they significantly populate alternative, locally unfolded conformations. The results, therefore, provide a view of the locations of low lying, locally unfolded conformations. They indicate approximately 4-6 local minima for each protein, all within ca. 2.5 kcal/mol of the native state, implying a locally rough energy landscape. Comparison with folding data for these proteins shows that folding involves most molecules traversing a similar path, once a transition state containing a beta hairpin has been formed, thereby defining a well-populated pathway down the folding funnel. The hairpin that directs the folding pathway differs for the two proteins and remains the most stable part of the folded protein. PMID- 16271892 TI - The binding mode of the trigger factor on the ribosome: implications for protein folding and SRP interaction. AB - This study presents the X-ray structure of the N-terminal binding domain of the D. radiodurans trigger factor (TF) in complex with the D. radiodurans large ribosomal subunit. At 3.35 A, a complete description of the interactions with ribosomal proteins L23, L29, and 23S rRNA are disclosed, many of which differ from those found previously for a heterologous bacterial-archaeal TF-ribosome complex. The beta hairpin loop of eubacterial L24, which is shorter in archaeal ribosomes, contacts the TF and severely diminishes the molecular cradle proposed to exist between the TF and ribosome. Bound to the ribosome, TF exposes a hydrophobic crevice large enough to accommodate the nascent polypeptide chain. Superimposition of the full-length TF and the signal-recognition particle (SRP) onto the complex shows that simultaneous cohabitation is possible, in agreement with biochemical data, and suggests a model for the interplay of TF, SRP, and the nascent chain during translation. PMID- 16271893 TI - Structure of the hepatitis C virus IRES bound to the human 80S ribosome: remodeling of the HCV IRES. AB - Initiation of translation of the hepatitis C virus (HCV) polyprotein is driven by an internal ribosome entry site (IRES) RNA that bypasses much of the eukaryotic translation initiation machinery. Here, single-particle electron cryomicroscopy has been used to study the mechanism of HCV IRES-mediated initiation. A HeLa in vitro translation system was used to assemble human IRES-80S ribosome complexes under near physiological conditions; these were stalled before elongation. Domain 2 of the HCV IRES is bound to the tRNA exit site, touching the L1 stalk of the 60S subunit, suggesting a mechanism for the removal of the HCV IRES in the progression to elongation. Domain 3 of the HCV IRES positions the initiation codon in the ribosomal mRNA binding cleft by binding helix 28 at the head of the 40S subunit. The comparison with the previously published binary 40S-HCV IRES complex reveals structural rearrangements in the two pseudoknot structures of the HCV IRES in translation initiation. PMID- 16271894 TI - Substrate-induced conformational changes in Bacillus subtilis glutamate racemase and their implications for drug discovery. AB - D-glutamate is an essential building block of the peptidoglycan layer in bacterial cell walls and can be synthesized from L-glutamate by glutamate racemase (RacE). The structure of a complex of B. subtilis RacE with D-glutamate reveals that the glutamate is buried in a deep pocket, whose formation at the interface of the enzyme's two domains involves a large-scale conformational rearrangement. These domains are related by pseudo-2-fold symmetry, which superimposes the two catalytic cysteine residues, which are located at equivalent positions on either side of the alpha carbon of the substrate. The structural similarity of these two domains suggests that the racemase activity of RacE arose as a result of gene duplication. The structure of the complex is dramatically different from that proposed previously and provides new insights into the RacE mechanism and an explanation for the potency of a family of RacE inhibitors, which have been developed as novel antibiotics. PMID- 16271895 TI - The N-terminal end of the catalytic domain of SRC kinase Hck is a conformational switch implicated in long-range allosteric regulation. AB - Signal transduction in cell growth and proliferation involves regulation of kinases through long-range allostery between remote protein regions. Molecular dynamics free energy calculations are used to clarify the coupling between the catalytic domain of Src kinase Hck and its N-terminal end connecting to the regulatory SH2 and SH3 modules. The N-terminal end is stable in the orientation required for the regulatory modules to remain properly bound only in the inactive catalytic domain. In the active catalytic domain, the N-terminal end prefers a different conformation consistent with dissociation of the regulatory modules. The free energy surface shows that the N-terminal end acts as a reversible two state conformational switch coupling the catalytic domain to the regulatory modules. Structural analogy with insulin receptor kinase and c-Src suggests that such reversible conformational switching in a critical hinge region could be a common mechanism in long-range allosteric regulation of protein kinase activity. PMID- 16271896 TI - A structure of the human apoptosome at 12.8 A resolution provides insights into this cell death platform. AB - Apaf-1 and cytochrome c coassemble in the presence of dATP to form the apoptosome. We have determined a structure of the apoptosome at 12.8 A resolution by using electron cryomicroscopy and single-particle methods. We then docked appropriate crystal structures into the map to create an accurate domain model. Thus, we found that seven caspase recruitment domains (CARDs) form a central ring within the apoptosome. At a larger radius, seven copies of the nucleotide binding and oligomerization domain (NOD) associate laterally to form the hub, which encircles the CARD ring. Finally, an arm-like helical domain (HD2) links each NOD to a pair of beta propellers, which bind a single cytochrome c. This model provides insights into the roles of dATP and cytochrome c in assembly. Our structure also reveals how a CARD ring and the central hub combine to create a platform for procaspase-9 activation. PMID- 16271897 TI - Women's health: the struggle to restore hormonal balance. PMID- 16271898 TI - A rational approach to evidence gaps in the management of osteoporosis. AB - Major advancements in the treatment of osteoporosis have occurred over the last decade. Therapies including the anti-resorptive drugs such as alendronate and risedronate have been shown in randomized placebo-controlled trials to increase bone mineral density and reduce fracture risk. Anabolic therapy in the form of parathyroid hormone has been introduced as the first treatment to build bone mass. However, gaps in our knowledge about specific management issues that arise frequently among primary care providers persist. In this paper, three common clinical scenarios are discussed: a postmenopausal woman with only slightly reduced bone mineral density; an osteoporotic woman on anti-resorptive therapy for more than 5 years; and a woman who continues to fracture despite treatment. Evidence gaps in each treatment scenario are presented, and rational approaches to management are suggested. PMID- 16271899 TI - Prevention of osteoporotic fractures in the elderly. AB - Osteoporosis is a common and preventable disorder of the older adult skeleton that predisposes an individual to an increased risk of fracture, a major cause of disability in older adults. Most patients with osteoporosis have an identifiable cause of bone loss. Factors contributing to osteoporotic fractures are more often associated with disordered neuromuscular function affecting postural stability than disordered skeletal integrity. Effective pharmacologic agents are available for the prevention and treatment of osteoporosis. Prevention of osteoporotic fractures in the elderly, particularly nonvertebral fractures, presents unique challenges. Fracture prevention requires identification and management of disorders that contribute to falls, the prevention of falls, and reduction of the impact force of falls. Thus, both pharmacological and nonpharmacological strategies need to be employed. The presence of multiple co-morbidities further complicates management of osteoporosis in the elderly population. PMID- 16271900 TI - Three-day vs longer duration of antibiotic treatment for cystitis in women: systematic review and meta-analysis. AB - PURPOSE: We performed a meta-analysis to ascertain the efficacy and safety of the currently practiced 3-day antibiotic therapy for cystitis versus prolonged therapy (5 days or longer) to relieve symptoms and to achieve bacteriological cure. METHODS: The Cochrane Library, the Cochrane Renal Group's Register of trials, EMBASE and MEDLINE were searched to identify all randomized controlled trials comparing 3-day oral antibiotic therapy with prolonged therapy (5 days and longer) for uncomplicated cystitis in adult non-pregnant women. Two reviewers independently applied selection criteria, performed quality assessment, and extracted data. Relative risks (RR) with their 95% confidence intervals (CI) were estimated; a fixed effect model was used. An intention-to-treat analysis was performed whenever possible. RESULTS: Thirty-two trials and 9605 patients met inclusion criteria. For symptomatic failure rates no difference between 3-day and prolonged antibiotic regimens was found at short term (RR 1.16, 95% CI: 0.96 1.41) and long-term follow-up (RR 1.17, 95% CI: 0.99-1.38). Three-day treatment was less effective than prolonged therapy in preventing bacteriological failure, relative risk 1.37 (95% CI: 1.07-1.74) for short-term follow-up, and 1.47 (95% CI: 1.22-1.77) for long-term follow-up. Adverse effects were more common in the prolonged therapy group (RR 0.83, 95% CI: 0.79-0.91). The results were consistent for subgroup and sensitivity analyses. CONCLUSION: Antibiotic therapy for 3 days is similar to prolonged therapy in achieving symptomatic cure for cystitis, while the prolonged treatment is more effective in obtaining bacteriological cure. PMID- 16271901 TI - Injectable corticosteroids in treatment of arthritis of the knee. AB - Injection of corticosteroid preparations for the local treatment of musculoskeletal disorders affecting the knee, such as osteoarthritis and rheumatoid arthritis, has been used widely since the 1950s. This local therapy minimizes systemic toxicity and can result in rapid improvement in symptoms, most often as an adjunctive therapy for acute or severe symptom flares. The evidence assessing safety and efficacy is reviewed and analyzed. Intra-articular knee injection technique is described and is a procedure that is easy to learn and can be performed quickly in an office or hospital setting. PMID- 16271902 TI - Unsteady, unfocused, and unable to hear. PMID- 16271903 TI - What are these brown spots, doc? PMID- 16271904 TI - A pointed clue. PMID- 16271905 TI - Acute pain in the tip of the index finger. PMID- 16271906 TI - Frailty, hospitalization, and progression of disability in a cohort of disabled older women. AB - PURPOSE: To determine the association between a previously validated frailty phenotype and the development of new-onset dependence in activities of daily living, independent of hospitalizations and other established predictors of disability. SUBJECTS: Seven hundred and forty-nine women enrolled in the Women's Health and Aging Study-I who were independent in all activities in daily living when enrolled in the cohort. METHODS: Assessments and interviews were conducted through home visits at 6-month intervals for 3 years. Frailty was classified using a validated phenotype (> or =3 of the following: weight loss, exhaustion, slow walking, sedentariness, and weak grip), and hospitalizations were identified by self-report. Grouped-time proportional hazard models assessed associations among frailty, hospitalization, and the development of dependence in activities in daily living, adjusting for other factors. RESULTS: Twenty-five percent of the cohort (186/749) were frail at baseline; 56% (104/186) of frail versus 20% (23/117) of nonfrail women developed dependence in activities in daily living (P <.001). In multivariate analysis, frailty was independently associated with the development of dependence in activities in daily living (hazard ratio [HR] = 2.2; 95% confidence interval [CI]: 1.4 to 3.6), adjusting for hospitalization status, age, race, education, baseline functional status, cognition, depressive symptoms, number of chronic diseases, and self-reported health status. Additionally, a dose response relationship existed between the number of frailty criteria that a woman had and the hazard of subsequent dependence in activities in daily living. CONCLUSION: Frailty, conceptualized as an underlying vulnerability, and hospitalization, which marks an acute deterioration in health, were strongly and independently associated with new-onset dependence in activities in daily living. Additional research is needed to determine if dependence can be minimized by targeting resources and programs to frail older persons. PMID- 16271907 TI - The effect of hormone replacement on physical performance in community-dwelling elderly women. AB - PURPOSE: Our aim was to determine the effect of hormone replacement on physical performance measures, functional ability, physical activity, falls, and cognitive function in elderly women. SUBJECTS AND METHODS: Following a 3-month, open-label, run-in phase, we randomized 373 community-dwelling women aged 65 years and older to receive conjugated equine estrogen 0.625 mg/day plus or minus medroxyprogesterone 2.5 mg/day vs placebo for 3 years in a double-blind fashion. We assessed time to rise from a chair, timed walking, balance, Instrumental Activities of Daily Living, Physical Activity Scale of the Elderly, Folstein Mini Mental State Examination, and falls. RESULTS: Over 3 years, except for balance scores, performance measures declined significantly (all P<.05). There were no significant mean differences between women on hormone replacement versus placebo for rising time (0.1, -0.5 to 0.7 seconds [mean, 95% confidence interval]), walking normal (0.0, -0.4 to 0.4 seconds), and walking fast (0.2, -0.1 to 0.6 seconds). There were no significant mean differences between the two groups for the Instrumental Activities of Daily Living (0.1, -0.1 to 0.3 points), Physical Activity Scale of the Elderly (-3, -15 to 8 points), Folstein Mini-Mental State Examination (-0.1, -0.3 to 0.3 points), or the proportion of participants reporting falls (-1, -11 to 9%). CONCLUSION: In elderly women, hormone replacement had no statistically significant effect on cognition or balance, nor did it prevent the age-related decline in physical measures of mobility, ability to rise from a chair, self-reported activities of daily living, physical activity scores, or falls. PMID- 16271908 TI - Prescription of teratogenic medications in United States ambulatory practices. AB - PURPOSE: The purpose of this study was to identify the potentially teratogenic medications most frequently prescribed to women of childbearing age and the specialty of physicians who provide ambulatory care to women who use such medications. In addition, we evaluated rates of contraceptive counseling to explore awareness of the risks associated with teratogenic medication use. SUBJECTS AND METHODS: The prescription of teratogenic medications and provision of contraceptive counseling on 12,681 visits made by nonpregnant women, 14 to 44 years of age, to 1880 physicians in US ambulatory practice (National Ambulatory Medical Care Survey) between 1998 and 2000 was analyzed. RESULTS: Use of a potentially teratogenic, class D or X, medication by a woman of childbearing age is documented on 1 of every 13 visits made to US ambulatory practices. These include anxiolytics (4.1 million annual prescriptions), anticonvulsant medications (1.4 million annual prescriptions), antibiotics like doxycycline (1.4 million annual prescriptions), and statins (0.8 million annual prescriptions). Isotretinoin accounts for less than 5% of potentially teratogenic prescriptions (0.5 million annual prescriptions). Internists and family/general practitioners provide ambulatory care to 45% of women prescribed potentially teratogenic medications, psychiatrists provide ambulatory care to 20% of women prescribed potentially teratogenic medications, and dermatologists provide ambulatory care to 20% of women prescribed potentially teratogenic medications. Contraceptive counseling was provided on less than 20% of visits that documented use of a potential teratogen by a woman of childbearing age. Women using low-risk (class A or B) drugs received contraceptive counseling as frequently as women using potential teratogens (P = .24). CONCLUSION: Potentially teratogenic medications are prescribed to millions of women of childbearing age each year. Physician awareness of the teratogenic risk associated with class D or X medications seems low. PMID- 16271909 TI - Homocysteine as a predictive factor for hip fracture in elderly women with Parkinson's disease. AB - PURPOSE: Incidence of hip fractures among elderly patients with Parkinson's disease is high. Recent studies have found that levodopa induces hyperhomocysteinemia in Parkinson's disease. Hyperhomocysteinemia is considered to be a risk factor for osteoporotic fractures in elderly men and women. Very high plasma homocysteine levels are a feature of homocystinuria, characterized by the early onset of osteoporosis. To determine the association between plasma homocysteine concentration and the risk of hip fracture in Parkinson's disease patients receiving levodopa, we prospectively studied a cohort of elderly women with Parkinson's disease. METHODS: We studied 199 elderly women with Parkinson's disease receiving levodopa therapy, from whom blood samples had been obtained to measure plasma homocysteine. Age-adjusted incidence rates of hip fractures were calculated for quartiles of plasma homocysteine concentrations. Cox proportional hazard regression was used to calculate hazard ratios for quartiles of homocysteine values. RESULTS: The mean duration of follow-up was 4.9 years. Hip fractures occurred in 66 patients. The age-adjusted incidence rates per 1000 person-years for hip fractures, from the lowest to the highest quartile of plasma homocysteine levels, were 1.59 (95% confidence interval [CI], 1.01-2.24), 1.57 (95% CI, 0.98-2.19), 1.21 (95% CI, 0.61-1.72), and 26.98 (95% CI, 16.48-37.24). The risk of hip fractures was greater in the highest quartile than that in the lowest, and the risk was almost 2.4 times higher. CONCLUSION: These findings suggest that the homocysteine concentration is an important risk factor for hip fractures in Parkinson's disease patients receiving levodopa. PMID- 16271910 TI - Gender and perceived severity of cardiac disease: evidence that women are "tougher". AB - PURPOSE: Patients' beliefs about their disease may affect their willingness to engage in preventive health behaviors. We sought to determine whether men and women with acute coronary syndrome differ in their perceptions of the severity of cardiac-related illness while controlling for the clinical severity of their condition. METHODS: All patients with acute coronary syndrome discharged from a university hospital during a 3-year period were mailed a questionnaire, and medical records were abstracted. The questionnaire assessed perceived severity of cardiac-related illness (5-point scale from "very mild" to "very severe"), symptom frequency, type of acute coronary syndrome event, number of medications, Duke Activity Status Index (DASI), time since most recent cardiac event, Charlson Comorbidity Index, and demographic information. A logistic regression model was constructed with perceived severity of heart disease as the dependent variable. Gender was the key independent variable while controlling for the other patient and disease variables. RESULTS: The 490 respondents (1217 surveys sent, 40.3% response rate) included 348 men and 142 women who were similar with regard to race and type of acute coronary syndrome event experienced. Women were older, less educated, had a lower DASI score, had more symptoms, and were taking more medications. However, they perceived their cardiac disease as being no more severe than the men. The significant predictors in the regression model of perceived severity included gender, DASI, number of symptoms, type of acute coronary syndrome event, and comorbidity. Female gender was associated with lower perceived severity (odds ratio 0.30-0.80). CONCLUSIONS: Women rate their cardiac disease as less severe than do men when controlling for other measures of cardiac disease severity. PMID- 16271911 TI - Impact of a general practitioner educational intervention on osteoarthritis treatment in an elderly population. AB - PURPOSE: We examined whether a continuing medical education intervention increased general practitioners' ability to select the proper pharmacological treatment for patients with osteoarthritis. SUBJECTS AND METHODS: Eight towns in Quebec, Canada were randomly allocated to one of four intervention options, workshop and decision tree, workshop, decision tree, or no intervention. All general practitioners practicing in each town were eligible to participate. We evaluated all dispensed prescriptions for either a cyclooxygenase (COX)-2 inhibitor, nonselective nonsteroidal anti-inflammatory drug or acetaminophen written by eligible general practitioners between May 2000 and June 2001 to elderly patients suffering from osteoarthritis. We used a multi-level Bayesian hierarchical model to assess the impact of the interventions on prescription adequacy. RESULTS: We analyzed 5318 dispensed prescriptions written by 249 general practitioners in the five-month preintervention period and 4610 dispensed prescriptions written by the same physicians in the five-month postintervention period. A score of zero or one was given to every prescription, with one indicating prescription adequacy according to guidelines provided during the interventions. Bayesian hierarchical models showed some improvement in scores in the post- versus preintervention periods in all four groups. The probability of an improvement in the towns allocated the workshop and decision tree over the control was 94%, compared with 74% in the workshop group and 55% in the decision tree group. CONCLUSION: An interactive approach offered by peers and complemented by easy to use guidelines may enhance the general practitioner's ability to manage osteoarthritis patients. PMID- 16271912 TI - Celecoxib compared with lansoprazole and naproxen to prevent gastrointestinal ulcer complications. AB - PURPOSE: Selective cyclooxygenase-2 (COX-2) inhibitors cause significantly fewer peptic ulcers than conventional nonselective nonsteroidal anti-inflammatory drugs (NSAIDs) in patients at low risk or high risk for peptic ulcers. On the other hand, proton pump inhibitor co-therapy has also been shown to be effective in preventing relapse of peptic ulcers in high-risk patients using nonselective NSAIDs. We compared the efficacy of a selective COX-2 inhibitor with that of proton pump inhibitor co-therapy in the reduction in the incidence of ulcer relapse in patients with a history of NSAID-related peptic ulcers. MATERIALS AND METHODS: For this study, we recruited 224 patients who developed ulcer complications after NSAID use. We excluded patients who required concomitant aspirin treatment and who had renal impairment. After healing of ulcers and eradication of Helicobacter pylori, patients were randomly assigned to treatment with celecoxib 200 mg daily (n = 120) or naproxen 750 mg daily and lansoprazole 30 mg daily (n = 122) for 24 weeks. The primary endpoint was recurrent ulcer complications. RESULTS: During a median follow-up of 24 weeks, 4 (3.7%, 95% confidence interval [CI] 0.0%-7.3%) patients in the celecoxib group, compared with 7 patients (6.3%, 95% CI 1.6%-11.1%) in the lansoprazole group, developed recurrent ulcer complications (absolute difference -2.6%; 95% CI for the difference -9.1%-3.7%). Celecoxib was statistically non-inferior to lansoprazole co-therapy in the prevention of recurrent ulcer complications. Concomitant illness (hazard ratio 4.72, 95% CI 1.24-18.18) and age 65 years or more (hazard ratio 18.52, 95% CI 2.26-142.86) were independent risk factors for ulcer recurrences. Significantly more patients receiving celecoxib (15.0%, 95% CI 9.7 22.5) developed dyspepsia than patients receiving lansoprazole (5.7%, 95% CI 2.8 11.4. P = .02). CONCLUSIONS: Celecoxib was as effective as lansoprazole co therapy in the prevention of recurrences of ulcer complications in subjects with a history of NSAID-related complicated peptic ulcers. However, celecoxib, similar to lansoprazole co-therapy, was still associated with a significant proportion of ulcer complication recurrences. In addition, more patients receiving celecoxib developed dyspepsia than patients receiving lansoprazole and naproxen. PMID- 16271913 TI - What to wear today? Effect of doctor's attire on the trust and confidence of patients. AB - PURPOSE: There are very few studies about the impact of physicians' attire on patients' confidence and trust. The objective of this study was to determine whether the way a doctor dresses is an important factor in the degree of trust and confidence among respondents. METHODS: A cross-sectional descriptive study using survey methodology was conducted of patients and visitors in the waiting room of an internal medicine outpatient clinic. Respondents completed a written survey after reviewing pictures of physicians in four different dress styles. Respondents were asked questions related to their preference for physician dress as well as their trust and willingness to discuss sensitive issues. RESULTS: Four hundred respondents with a mean age of 52.4 years were enrolled; 54% were men, 58% were white, 38% were African-American, and 43% had greater than a high school diploma. On all questions regarding physician dress style preferences, respondents significantly favored the professional attire with white coat (76.3%, P <.0001), followed by surgical scrubs (10.2%), business dress (8.8%), and casual dress (4.7%). Their trust and confidence was significantly associated with their preference for professional dress (P <.0001). Respondents also reported that they were significantly more willing to share their social, sexual, and psychological problems with the physician who is professionally dressed (P <.0001). The importance of physician's appearance was ranked similarly between male and female respondents (P=.54); however, female physicians' dress appeared to be significantly more important to respondents than male physicians' dress (P <.001). CONCLUSION: Respondents overwhelmingly favor physicians in professional attire with a white coat. Wearing professional dress (ie, a white coat with more formal attire) while providing patient care by physicians may favorably influence trust and confidence-building in the medical encounter. PMID- 16271914 TI - Obstructive intramural coronary amyloidosis and myocardial ischemia are common in primary amyloidosis. AB - PURPOSE: The purpose of this study was to determine the prevalence of clinical syndromes and pathologic changes of myocardial ischemia due to obstructive intramural coronary amyloidosis among patients with primary amyloidosis and cardiac involvement. SUBJECTS AND METHODS: Medical records and pathologic specimens were reviewed from 96 patients with primary amyloidosis and cardiac involvement at autopsy or after cardiac transplantation during a 20-year period. Medical records were reviewed for patient demographic and clinical characteristics, including evidence for syndromes of myocardial ischemia. Pathologic specimens were examined for obstructive intramural coronary amyloidosis and microscopic changes of myocardial ischemia. RESULTS: Obstructive intramural coronary amyloidosis was present in 63 of 96 patients (66%). Microscopic changes of myocardial ischemia were more common in patients with obstructive intramural coronary amyloidosis (86%) than in those without (52%) (P <.001). In the 76 patients without coexistent severe epicardial coronary atherosclerosis, changes of myocardial ischemia were more common in those with obstructive intramural coronary amyloidosis (83%) than in those without (45%) (P <.001). In patients who had tissue available for review, none had obstruction of epicardial coronary arteries from amyloid. Syndromes of myocardial ischemia affected 16 patients (25%) with obstructive intramural coronary amyloidosis but only 2 patients (6%) without (P=.027). For 11% of the patients with obstructive intramural coronary amyloidosis, a syndrome of myocardial ischemia consisting of acute myocardial infarction or angina pectoris was the first manifestation of primary amyloidosis. CONCLUSION: Most patients with primary systemic amyloidosis and cardiac involvement have obstructive intramural coronary amyloidosis and associated microscopic changes of myocardial ischemia. Syndromes of myocardial ischemia may occur in these patients. PMID- 16271915 TI - Spontaneous pyogenic vertebral osteomyelitis and endocarditis: incidence, risk factors, and outcome. AB - PURPOSE: The relationship between pyogenic vertebral osteomyelitis and infectious endocarditis is uncertain. This study investigates the incidence and risk factors of infectious endocarditis in patients with pyogenic vertebral osteomyelitis, and the outcome of pyogenic vertebral osteomyelitis with and without associated infectious endocarditis. METHODS: A retrospective record review was conducted of all cases of vertebral osteomyelitis from January 1986 to June 2002, occurring in a tertiary referral hospital. Patients were followed for at least 6 months with careful attention to detection of infectious endocarditis and relapses. RESULTS: Among 606 patients with infectious endocarditis, 28 (4.6%) had pyogenic vertebral osteomyelitis. Among 91 cases of pyogenic vertebral osteomyelitis, 28 (30.8%) had infectious endocarditis. In 6 patients with no clinical signs of infectious endocarditis, the disease was established by routine echocardiography. Infectious endocarditis was more common in patients with predisposing heart conditions and streptococcal pyogenic vertebral osteomyelitis infection. Overall, pyogenic vertebral osteomyelitis in-hospital mortality was 11% (7.1% with infectious endocarditis). Twelve of 25 patients with infectious endocarditis with uncomplicated pyogenic vertebral osteomyelitis were treated for 4 to 6 weeks (endocarditis protocol), with no pyogenic vertebral osteomyelitis relapses. CONCLUSIONS: When specifically sought, the incidence of infectious endocarditis is high in patients with pyogenic vertebral osteomyelitis. Oral therapy may be an option for uncomplicated pyogenic vertebral osteomyelitis; nevertheless, in gram positive infections, this approach should only be considered after excluding infectious endocarditis. Favorable outcome with shorter treatment in uncomplicated pyogenic vertebral osteomyelitis associated with infectious endocarditis suggests that prolonged therapy may not be needed in this subgroup except for those infected by difficult to treat microorganisms, such as methicillin-resistant Staphylococcus aureus or Candida spp. PMID- 16271916 TI - Use of N-terminal pro-B-type natriuretic peptide to detect myocardial ischemia. AB - PURPOSE: To evaluate the utility of plasma N-terminal pro-B-type natriuretic peptide (NT-proBNP) levels to detect myocardial ischemia. METHODS: We conducted a prospective observational study in 260 consecutive patients with suspected myocardial ischemia referred for rest/ergometry myocardial perfusion single photon emission computed tomography. Levels of NT-proBNP were determined before and immediately after symptom-limited bicycle ergometry. RESULTS: Inducible myocardial ischemia on perfusion images was detected in 129 patients (49.6%). Baseline NT-proBNP and exercise induced increase in NT-proBNP (DeltaNT-proBNP) were significantly higher in patients with myocardial ischemia (median baseline NT-proBNP 155 pg/mL vs 91 pg/mL, P <.001; DeltaNT-proBNP 15 pg/mL vs 7 pg/mL, P = .002). Compared with patients in the lowest DeltaNT-proBNP quartile, those in the highest quartile of DeltaNT-proBNP had three times the risk of inducible ischemia (relative risk, 2.9; 95% confidence interval, 1.4 to 6.0; P = .003). Overall, the accuracy of baseline NT-proBNP and DeltaNT-proBNP in the detection of myocardial ischemia were similar to that of the exercise electrocardiogram (ECG). Combining exercise ECG and baseline NT-proBNP or DeltaNT-proBNP slightly increased the accuracy of exercise ECG only. CONCLUSION: The NT-proBNP level at rest as well as DeltaNT-proBNP during exercise stress testing is associated with inducible myocardial ischemia. NT-proBNP levels may have incremental value in the diagnosis of myocardial ischemia. PMID- 16271917 TI - Aspirin use for the primary prevention of coronary heart disease in older adults. AB - PURPOSE: Aspirin for the primary prevention of coronary heart disease (has a more favorable risk/benefit profile among adults with high coronary heart disease risk than among low-risk adults, but there is little information on the current patterns of aspirin use for primary prevention. We determined the prevalence of aspirin use in relation to coronary heart disease risk and changes over time. SUBJECTS AND METHODS: We measured regular aspirin use in 2163 black and white older adults without cardiovascular disease in a population-based cohort from 1997 to 1998 and 2002 to 2003. We determined the 10-year coronary heart disease risk by using the Framingham risk score. RESULTS: In 1997-1998, 17% of the cohort were regular aspirin users. Aspirin use increased with coronary heart disease risk from 13% in persons with a 10-year risk less than 6% (low risk) to 23% in those with a 10-year risk greater than 20% (highest risk) (P for trend < .001). Blacks were less likely to use aspirin (13%) than whites (20%). In multivariate analysis, black race was still associated with lower aspirin use (odds ratio 0.66, 95% confidence interval 0.49-0.89). In 1997-1998 and 2002 to 2003, aspirin use increased from 17% to 32% among those still free of coronary heart disease (P < .001), and the association with coronary heart disease risk continued (P for trend < .001). Despite their high coronary heart disease risk, diabetic persons were not more likely to use aspirin than nondiabetic persons, even in 2002 and 2003 (odds ratio 0.89, 95% confidence interval 0.56-1.40). CONCLUSION: Regular use of aspirin by older adults with no history of cardiovascular disease has increased in recent years. Individuals at higher coronary heart disease risk are more likely to take aspirin, but there is room for considerable improvement in targeting those at high risk, particularly diabetic persons and blacks. PMID- 16271918 TI - Proinflammatory state and circulating erythropoietin in persons with and without anemia. AB - PURPOSE: High circulating levels of proinflammatory cytokines cause anemia, perhaps by interacting with erythropoietin production or biological activity. We characterize the relationships of systemic inflammation, erythropoietin, and hemoglobin. METHODS: Data are from the InCHIANTI (Invecchiare in Chianti, aging in the Chianti area) study population. A sample of 1270 persons aged 65 years or older and 30 men and 30 women from each age-decade 20 to 70 years were randomly selected from the residents in the Chianti, Italy, geographic area. Of the 1714 eligible persons, 1235 had complete data on inflammatory markers, erythropoietin, hemoglobin, potential causes of anemia, and other relevant covariates. Anemia was defined as hemoglobin less than 12 g/dL in women and less than 13 g/dL in men. RESULTS: Independent of age, sex, and hemoglobin, the number of elevated inflammatory markers (C-reactive protein, interleukin-6, interleukin-1beta, and tumor necrosis factor-alpha) was associated with progressively higher erythropoietin in non-anemic participants but lower erythropoietin in anemic participants. Findings were consistent across different causes of anemia. The threshold at which the effect of inflammation on erythropoietin reversed was close to 13.0 g/dL of hemoglobin. CONCLUSIONS: Our findings suggest that anemia of inflammation evolves from a "pre-anemic" stage characterized by a compensatory increment of erythropoietin that maintains normal hemoglobin levels to a stage of clinically evident anemia in which erythropoietin levels are not high enough to maintain normal hemoglobin, possibly because of the inhibitory effect of inflammation on erythropoietin production. This hypothesis requires testing in a longitudinal study. PMID- 16271919 TI - Nephrotic range proteinuria and CD4 count as noninvasive indicators of HIV associated nephropathy. AB - PURPOSE: Human immunodeficiency virus (HIV)-associated nephropathy is a common and serious cause of progressive renal insufficiency in patients with HIV, frequently presenting with nephrotic range proteinuria. The purpose of this study is to document the histopathologic diagnoses seen in HIV-positive patients with and without nephrotic range proteinuria and to evaluate the predictive value of both nephrotic range proteinuria and CD4 count in diagnosing HIV-associated nephropathy. METHODS: We performed a cross-sectional, single-center study of all 107 HIV-positive patients who had both a renal biopsy and urine protein measurement between 1995 and 2002. Nephrotic range proteinuria was defined as a urine protein-to-creatinine ratio > 3 or a 24-hour urine protein > 3 g. Clinical and laboratory characteristics of those patients with and without HIV-associated nephropathy were compared. Sensitivity, specificity, and positive and negative predictive values of nephrotic range proteinuria in the diagnosis of HIV associated nephropathy were determined. RESULTS: Fifty-five biopsied patients had nephrotic range proteinuria, among whom 29 (53%) were diagnosed with HIV associated nephropathy. Among the remaining patients, 12 had non-HIV-associated nephropathy focal segmental glomeruloscerlosis, 3 had membranoproliferative glomerulonephritis, 2 had AA Amyloid, 2 had diabetic nephropathy, and 7 had other diagnoses. Sensitivity, specificity, and positive and negative predictive values of nephrotic proteinuria in the diagnosis of HIV-associated nephropathy were 73%, 61%, 53%, and 79%, respectively. The patients with HIV-associated nephropathy had a significantly higher creatinine (8.2 mg/dL vs 2.5 mg/dL, P < .001) and a lower CD4 count (158 count/mm3 vs 349 count/mm3, P < .01) at the time of biopsy. Although significantly more patients with HIV-associated nephropathy had a CD4 count below 200 (P = .03), among those with a CD4 count below 200, 10 of 30 patients (33%) had diagnoses other than HIV-associated nephropathy. Injection drug use, presence of hepatitis C, and hypertension were not associated with HIV associated nephropathy. CONCLUSION: Our results suggest that HIV patients with nephrotic range proteinuria warrant a kidney biopsy because the presence of nephrotic range proteinuria, even in the presence a low CD4 count, does not establish the diagnosis of HIV-associated nephropathy. PMID- 16271920 TI - A randomized trial comparing electronic and conventional stethoscopes. PMID- 16271921 TI - Acarbose attenuates the hypotensive response to sucrose and slows gastric emptying in the elderly. PMID- 16271922 TI - Avoiding the winner's curse in faculty recruitment. PMID- 16271923 TI - Female genital tuberculosis: uncommon presentation of tuberculosis in the United States. PMID- 16271924 TI - Disseminated nocardiosis with initial manifestation mimicking disease flare-up of systemic lupus erythematosus in an SLE patient. PMID- 16271925 TI - Hemophagocytosis in the peripheral blood due to tuberculosis mycobacteremia. PMID- 16271926 TI - Restrictive ventilatory dysfunction and dyspnea in elderly subjects. PMID- 16271928 TI - Dyspnea in the elderly. PMID- 16271929 TI - Teaching clinical competence. PMID- 16271932 TI - Bedside teaching of medical students. PMID- 16271933 TI - Non-invasive carbon dioxide pressure measurement. PMID- 16271937 TI - What can poetry teach physicians? PMID- 16271935 TI - White cell differential count and influenza A. PMID- 16271938 TI - Diminazene aceturate resistance on the virulence of Trypanosoma brucei for rats. AB - Four groups (A, B, C and D) of 10 naive rats were used to compare the virulence of isolates of a strain of Trypanosoma brucei before and after the development of diminazene aceturate resistance. Group A rats were uninfected (controls). Group B rats were infected with a trypanosome isolate unexposed to the drug, while groups C and D rats were infected with two different drug-resistant isolates of the same strain. Rats in the three infected groups each received 10(6) trypanosomes intraperitoneally. Prepatent periods were longer (P<0.05) in groups C and D than in group B. The parasitaemic periods in groups B, C and D were similar, but group C and D rats differed from group B rats in surviving longer, and in showing lower degrees of parasitaemia, anaemia and hepatomegaly. No differences were noted between group C and D rats. Thus, diminazene aceturate resistance appeared to reduce the virulence of T. brucei. PMID- 16271939 TI - Telmisartan inhibits expression of a receptor for advanced glycation end products (RAGE) in angiotensin-II-exposed endothelial cells and decreases serum levels of soluble RAGE in patients with essential hypertension. AB - There is a growing body of evidence that the advanced glycation end product (AGE) their receptor (RAGE) system plays a central role in the pathogenesis of diabetic vascular complication. The renin-angiotensin system (RAS) contributes to the development and progression of diabetic angiopathy as well. However, the cross talk between the AGE-RAGE system and the RAS is not fully understood. In this study, we examined the role of angiotensin II (Ang II) type 1 receptor system for RAGE expression in cultured endothelial cells (ECs) and in patients with essential hypertension. Ang II up-regulated RAGE mRNA levels of microvascular ECs and subsequently increased the soluble form of RAGE (sRAGE) expression in the medium of ECs, both of which were completely blocked by telmisartan, a commercially available Ang II type 1 receptor antagonist. Furthermore, telmisartan was found to decrease serum levels of sRAGE in patients with essential hypertension. These results demonstrate that sRAGE is released from the cell surface of Ang-II-exposed ECs. Our present study indicates that a cross-talk exists between the AGE-RAGE system and the RAS and suggests that serum levels of sRAGE may reflect endothelial RAGE expression. PMID- 16271940 TI - Transport properties of the mesothelium and interstitium measured in rabbit pericardium. AB - The contribution of the pleural mesothelium to pleural liquid and protein transport is still vigorously debated. Recent in vitro studies of stripped pleural membrane and free-standing pericardium have demonstrated active ion solute coupled transport of liquid and transcytosis of protein. However, the relative contribution of the passive transport properties of the pleural mesothelium compared to the pleural interstitium has not been extensively studied. In in vitro studies, we measured the albumin diffusion coefficient, reflection coefficient, hydraulic conductivity and electrical resistance of rabbit pericardium. We used two techniques, treatment with 40 muM nocodazole and a 1-min hypotonic cell lysis with distilled water, to eliminate the effect of the two mesothelial layers on diffusional and hydraulic resistances. Each technique increased the albumin diffusion coefficient and hydraulic conductivity 3- to 4 fold. In hydraulic conductivity experiments using tracer 125I-albumin, nocodazole reduced the reflection coefficient to zero, rendering the pericardium completely permeable to albumin. We applied the cell-lysis technique to the pleural and pericardial mesothelium in sequence to evaluate the separate contribution of each mesothelium. Both diffusional and hydraulic resistances, but not electrical resistance, of the mesothelium were overestimated by the cell-lysis technique. The pleural mesothelium contributed at most 30% of diffusional resistance, 10% of hydraulic resistance and 14% of electrical resistance of the total pericardial resistances. We conclude that the pleural mesothelium is not the primary barrier to protein diffusion or bulk flow of liquid from the pericardial microcirculation to the pleural liquid. PMID- 16271941 TI - Vascular endothelial growth factor receptor-2 and low affinity VEGF binding sites on human glomerular endothelial cells: Biological effects and advanced glycosilation end products modulation. AB - Vascular Endothelial Growth Factor (VEGF), binding to its receptor in endothelial cells, seems to modulate the increased blood flow in the early phase of diabetic renal disease. The aim of the study was to evaluate, in a diabetic milieu, the expression, biological function and modulation of VEGF binding sites in human glomerular endothelial cells (GENC). We demonstrated the presence of VEGF binding sites with high (VEGFR-2) and low (heparan sulfate proteoglycans, HSPG) affinity. VEGF165 and VEGF121 working through VEGFR-2 stimulated nitric oxide (NO) production at low doses (0.1-1 nM), whereas only VEGF165 at high doses (10-100 nM) increased thymidine incorporation. 1 nM VEGF165 and VEGF121 induced in GENC a significant peak of inducible NO synthase (iNOS) production and, at a lower level, of endothelial NOS (eNOS). The copresence of VEGF165 with aminoguanidine (iNOS inhibitor) determined an increase of eNOS and a significant increase in thymidine incorporation. Advanced glycation end products (AGEs) working through specific receptors (RAGE) up-regulated the expression of VEGFR-2, decreased the expression of HSPG sites and reduced GENC growth. These results identify in GENC VEGFR-2 as a mediator of iNOS and eNOS release under control of VEGF, whereas HSPG binding sites seem to mediate the weak growth effect. The presence of AGEs, up-regulating the VEGFR-2 and decreasing HSPG sites might participate to the block of glomerular angiogenesis addressing the VEGF effects on glomerular permeability. PMID- 16271942 TI - Electrical and structural remodeling: role in the genesis and maintenance of atrial fibrillation. AB - Atrial fibrillation (AF) and congestive heart failure (CHF) are 2 frequently encountered conditions in clinical practice. Both lead to changes in atrial function and structure, an array of processes known as atrial remodeling. This review provides an overview of ionic, electrical, contractile, neurohumoral, and structural atrial changes responsible for initiation and maintenance of AF. In the last decade, many studies have evaluated atrial remodeling due to AF or CHF. Both conditions often coexist, which makes it difficult to distinguish the contribution of each. Because of atrial stretch in the setting of hypertension or CHF, atrial remodeling frequently occurs long before AF arises. Alternatively, AF may lead to electrical remodeling, that is, shortening of refractoriness due to the high atrial rate itself. In many experimental AF or rapid atrial pacing studies, the ventricular rate was uncontrolled. In those studies, atrial stretch due to CHF may have interfered with the high atrial rate to produce a mixed type of electrical and structural remodeling. Other studies have dissected the individual role of AF or atrial tachycardia from the role CHF plays in atrial remodeling. Atrial fibrillation itself does not lead to structural remodeling, whereas this is frequently produced by hypertension or CHF, even in the absence of AF. Primary and secondary prevention programs should tailor treatment to the various types of remodeling. PMID- 16271943 TI - Surgical ablation of atrial fibrillation. AB - Surgical ablation of atrial fibrillation (AF) is the most effective means of curing this arrhythmia, with the classic Maze procedure eliminating AF in more than 90% of patients. A complex but safe operation, the Maze procedure has been applied by relatively few surgeons. However, recent advances in the understanding of the pathogenesis of AF and development of new ablation technologies enable surgeons to perform pulmonary vein ablation, create linear left atrial lesions, and remove the left atrial appendage rapidly and safely. Lesions are created under direct vision, minimizing the risk of damage to the pulmonary veins and adjacent mediastinal structures. Most surgical ablation procedures have been performed in conjunction with mitral valve surgery, the combination of mitral valve repair and cure of AF enabling patients to avoid lifelong anticoagulation. Recently developed surgical instrumentation now enables thoracoscopic and keyhole approaches, facilitating extension of epicardial AF ablation and excision of the left atrial appendage to patients with isolated AF and no other indication for cardiac surgery. PMID- 16271944 TI - Catheter ablation therapy for atrial fibrillation. AB - Since cases were first reported in 1994, catheter ablation of atrial fibrillation has undergone rapid development and expansion. The procedure began as an attempt to recreate the Maze III operation with a catheter technique. Understanding the contribution of the pulmonary veins to the initiation and maintenance of atrial fibrillation led to dramatic changes in procedural technique. The segmental ostial and the circumferential approaches have emerged as the 2 dominant methods. Efforts continue in academic centers to better understand the pathophysiology of the arrhythmia and to further refine the ablation procedure to improve patient outcomes. PMID- 16271945 TI - New antiarrhythmic drugs for prevention of atrial fibrillation. AB - Enhanced understanding of the mechanisms underlying atrial fibrillation (AF) and advent of catheter-based therapy for AF has altered the approach to patients with this most common arrhythmia. However, despite the success of aggressive procedural techniques, pharmacologic therapy remains the first-line and mainstay approach in the treatment of AF. This review of new antiarrhythmic drug (AAD) therapy for AF provides an in-depth overview of recently available classic and new investigational drugs being considered for AF treatment. Currently available AADs are associated with less than satisfactory efficacy in preventing AF and a significant side effect profile, including ventricular proarrhythmia. Recent investigations have focused on development of new AADs that, hopefully, will be more effective and safer even in patients with structural heart disease. These new AADs include selective multi-ion channel and atrial specific blockers and agents that target the underlying etiologies and substrate alterations that lead to AF. Included among the latter new category are agents that suppress activation of the renin-angiotensin-aldosterone system or inflammation, which represent novel targets for drug therapy for AF. Finally, new selective A1 adenosine receptor agonists may offer the possibility of more specific and successful ventricular rate control during AF. There is considerable hope and interest that improved understanding of AF mechanisms ultimately will result in more effective and less dangerous pharmacologic therapy becoming available in the future. PMID- 16271946 TI - Real-time 3-dimensional echocardiography: a review of the development of the technology and its clinical application. AB - Real-time 3-dimensional echocardiography (RT3DE) is a new imaging technique that can provide accurate, important, and additional information concerning cardiovascular morphology, pathology, and function. This article will review the development of the technology of RT3DE and its clinical application. As the technique continues to evolve, RT3DE is bound to play an increasingly important role in the diagnosis, prognosis, and treatment of patients with various forms of cardiovascular disease. PMID- 16271948 TI - Large cell transformation of mycosis fungoides: tetraploidization within skin tumor large cells. AB - Little is known about the molecular or cytogenetic alterations of mycosis fungoides (MF) large cell transformation. We report our findings on chromosomal rearrangement, based on peripheral blood and skin examination before and after cutaneous MF large cell transformation, using both conventional and molecular cytogenetic techniques. Blood cells exhibited a similar hypodiploid karyotype before and after MF transformation. A near-tetraploid karyotype with complex structural rearrangements was established from a skin tumor after MF large cell transformation. Both recurrent chromosome abnormalities and an identical T-cell receptor gamma-chain rearrangement were shared by blood and skin cells, suggesting that MF large cell transformation derived from a common monoclonal ancestor detected at MF stage. A complex hypotetraploid karyotype was established only from the skin tumor, however. MF large cell transformation may be associated with chromosome duplication followed by chromosome losses and interchromosomal rearrangements. Accordingly, additional parallel blood and skin tumor cytogenetic studies are required to further identify the recurrent cytogenetic changes associated with the aggressiveness of the disease after large cell transformation. PMID- 16271949 TI - Smoking may cause genetic alterations at 5q22.2 approximately q23.1 in clear-cell renal cell carcinoma. AB - The aim of this study was to examine the relationship between allelic imbalance (AI) on chromosome 5q and clinico-pathologic parameters, including cigarette smoking. We examined the AI on chromosome 5q in 119 clear-cell renal cell carcinomas (CRCC) by a fluorescent polymerase chain reaction technique using nine microsatellite markers. AI of one or more loci was found in 43 cases (36.1%). The most frequent AI was observed at chromosome 5q23.1 (D5S467), where the LOX gene is located. The minimally involved region ranged from 5q22.1 to 5q23.2, with a possible breakpoint between 5q22 and 5q22.1. Regarding the relationships between the frequency of AI and the clinico-pathologic and environmental backgrounds, smokers had a significantly higher frequency of AI of 5q22.2 approximately q22.3 (D5S471) than nonsmokers (P = 0.013). No other significant associations were found between AI of specific loci and other parameters. Our results suggest that AI at 5q plays an important role in the genesis of CRCC. In addition, smoking may cause genetic alterations at 5q22.2 approximately q23.1, where the LOX gene is located. PMID- 16271950 TI - Random aneuploidy and telomere capture in chronic lymphocytic leukemia and chronic myeloid leukemia patients. AB - Telomeric regions of the human genome are of particular interest, because rearrangements of these regions are difficult to identify by conventional chromosome banding technology. With the advent of molecular cytogenetic techniques such as fluorescence in situ hybridization (FISH), it has been possible to investigate the terminus in cytogenetically visible terminal deletions and telomere rearrangements. We investigated telomere capture and aneuploidy rates in chronic lymphocytic leukemia (CLL) and chronic myeloid leukemia (CML) patients, as well as in healthy control subsets. Using a FISH technique, we estimated the random aneuploidy and telomere capture of the 21q22, SNRPN, and 15qter loci. Higher aneuploidy rates were found in the leukocytes of CLL and CML patients, compared with the control group, for the 21q22 and SNRPN loci. There was no difference in the aneuploidy rate between the CML and CLL groups. Telomere capture was found in the two groups (CLL and CML), but not in the control group. We propose that the telomere capture phenomenon is much more common than has been reported in the literature; however, its prognostic significance is yet to be established. PMID- 16271951 TI - Characterization of the TSU-PR1 cell line by chromosome painting and flow cytometry. AB - TSU-PR1 was originally reported as a prostatic carcinoma cell line derived from a lymph node metastasis. Recently, however, this cell line was reported to be derived from T24 bladder carcinoma cells, and thus further definition of its origin is needed. Conventional cytogenetic study of TSU-PR1 showed aneuploidy, ranging from 65 to 86 chromosome with a modal number of 80, and with 10 marker chromosomes, thus conventional cytogenetics cannot be used to determine which chromosomes or regions of chromosomes are critical in cancer development and progression of this cell line. The present study was conducted to characterize genetic changes of the cell line using comparative genomic hybridization (CGH), fluorescence in situ hybridization (FISH), and flow cytometry. CGH results showed that green-to-red fluorescence ratios were within the range of 0.85-1.15, except for a few chromosomes, which reflected near tetraploidy in TSU-PR1. Flow cytometric analysis of TSU-PR1 revealed a DNA index of 3.46n, which is close to the 3.48n calculated from a modal number of 80. The copy numbers of chromosomes 4, 6, 7, 17, and 20 determined by the DNA index and the CGH analyses were 2.85 +/ 0.09, 3.22 +/- 0.77, 3.01 +/- 0.26, 4.05 +/- 0.44, and 4.99 +/- 0.48, respectively. These numbers are also in accordance with the chromosome copy numbers determined with FISH: 2.98 +/- 0.23, 2.91 +/- 0.44, 2.74 +/- 0.44, 3.93 +/- 0.38, and 5.05 +/- 0.78 for chromosomes 4, 6, 7, 17, and 20, respectively (P > 0.05). PMID- 16271952 TI - Molecular characterization of the t(3;9) associated with immortalization in the MCF10A cell line. AB - The t(3;9)(p14;p21) in the MCF10A human mammary gland epithelial cell line was the single cytogenetic event that accompanied the transition from primary culture to immortalized cell line, suggesting that it is related to the development of the immortalization phenotype. To study the molecular consequences of the breakpoints in this rearrangement, we used a combination of fluorescence in situ hybridization (FISH) and array comparative genomic hybridization (CGH). The 3p14 translocation breakpoint occurs within BAC RP11-795e22, which accommodates only the TAFA1 gene, a novel cysteine-rich secreted protein thought to be involved in cytokine signaling. TAFA1 is expressed in normal breast tissue, not in MCF10A, and shows differential expression in a range of breast cancer cell lines. The 9p translocation breakpoint results in a deletion of approximately 4 megabases on the derivative chromosome 9, which includes the CDKN2A (p16) gene. Array CGH and FISH analysis demonstrated that BAC 149i22, which contains the CDKN2A/B genes, is also deleted specifically on the apparently normal copy of chromosome 9, making MCF10A null for the p16/p15 genes. The exact extent of gains and losses of chromosome regions resulting from rearrangements involving chromosomes 1q, 5q, and 8q have also been characterized using the BAC arrays. PMID- 16271953 TI - Immortalization of human extravillous cytotrophoblasts by human papilloma virus gene E6E7: sequential cytogenetic and molecular genetic characterization. AB - Extravillous cytotrophoblast (EVCT) cultures from the normal placentas of three pregnant women were transfected by HPVE6E7. Sequential cytogenetic and molecular analyses were performed to delineate genetic events that may be critical for cell immortalization. One line, PE1-E6E7, was immortalized successfully, whereas 2 other lines, PE3-E6E7 and PE4-E6E7, could not be maintained beyond crisis. Before crisis, the majority of cells in all lines were karyotypically normal. During the early stages of crisis, there was progressive telomere shortening. Most cells were karyotypically abnormal, with extreme cytogenetic divergence and a predominance of telomeric association and dicentric chromosomes affecting many chromosomes. At the later stages of crisis, the karyotype became more convergent with a drastic decrease in nonclonal aberrations. In PE1-E6E7, after crisis the karyotype was complex, with frequent centromeric rearrangements in the form of isochromosomes and whole-arm translocations. There were unbalanced structural aberrations and numerical changes, including loss of chromosome 13, that could be traced throughout the evolution of the line. These findings support the concept that immortalization is a relatively rare and nonrandom event that occurs only in cells that have acquired the necessary or critical genetic alterations. Telomeric dysfunction may be an important mechanism leading to the acquisition of complex karyotypical aberrations. PMID- 16271954 TI - The association of the DNA repair gene XRCC3 Thr241Met polymorphism with susceptibility to colorectal cancer in a Chinese population. AB - Growing evidence suggests that the Thr241Met (T241M) polymorphism in the homologous recombination repair gene XRCC3 may alter DNA repair capacity and subsequent susceptibility to carcinogens. In a few studies of colorectal cancer (CRC), however, the results have been discrepant. A population-based nested case control study including 140 cases and 280 cancer-free controls was conducted to evaluate the effect of XRCC3 polymorphism, environmental exposure, and family history (FH) on the risk of CRC. The variant allele frequency was low among the ethnic Han Chinese, but we observed a significant difference between cases (6.07%) and controls (2.32%). The analytic results of the unconditional logistic regression model adjusted by age, sex, alcohol intake, cigarette smoking, and FH of cancer in first-degree relatives showed a significantly increased risk of CRC (adjusted odds ratio [OR] = 3.13, 95% confidence interval [CI]: 1.41-6.95, P = 0.005) as the T/M and M/M genotypes compared with the T/T genotype, which changed weakly in consideration of the subsite (adjusted OR = 4.80, 95%CI: 1.77-12.98, P = 0.002 in colon cancer, adjusted OR = 2.41, 95%CI: 0.93-6.25, P = 0.071 in rectal cancer, respectively). Combined with environmental factors such as alcohol intake and cigarette smoking, no significant interaction could be found. However, the results revealed a significant association between FH of cancer in first degree relatives and the risk of CRC (adjusted OR = 2.24, 95%CI: 1.18-4.25, P = 0.014). These results also suggest that XRCC3 T241M polymorphism and FH of cancer may be risk factors for CRC, and the XRCC3 241Met allele may be an effective biomarker for genetic susceptibility to CRC. Larger studies are needed to confirm our findings and identify the underlying mechanisms. PMID- 16271955 TI - Chromosomal alterations cause the high rates and wide ranges of drug resistance in cancer cells. AB - Conventional mutation-selection theories have failed to explain (i) how cancer cells become spontaneously resistant against cytotoxic drugs at rates of up to 10(-3) per cell generation, orders higher than gene mutation, even in cancer cells; (ii) why resistance far exceeds a challenging drug-a state termed multidrug resistance; (iii) why resistance is associated with chromosomal alterations and proportional to their numbers; and (iv) why resistance is totally dependent on aneuploidy. We propose here that cancer-specific aneuploidy generates drug resistance via chromosomal alterations. According to this mechanism, aneuploidy varies the numbers and structures of chromosomes automatically, because it corrupts the many teams of proteins that segregate, synthesize, and repair chromosomes. Aneuploidy is thus a steady source of chromosomal variation from which, in classical Darwinian terms, resistance specific aneusomies are selected in the presence of chemotherapeutic drugs. Some of the thousands of unselected genes that hitchhike with resistance-specific aneusomies can thus generate multidrug resistance. To test this hypothesis, we determined the rates of chromosomal alterations in clonal cultures of human breast and colon cancer lines by dividing the fraction of nonclonal karyotypes by the number of generations of the clone. These rates were about 10(-2) per cell generation, orders higher than mutation. Chromosome numbers and structures were determined in metaphases hybridized with color-coded chromosome-specific DNA probes. Further, we tested puromycin-resistant subclones of these lines for resistance-specific aneusomies. Resistant subclones differed from parental lines in four to seven specific aneusomies, of which different subclones shared some. The degree of resistance was roughly proportional to the number of these aneusomies. Thus, aneuploidy is the primary cause of the high rates and wide ranges of drug resistance in cancer cells. PMID- 16271956 TI - Search for large genomic alterations of the BRCA1 gene in a Finnish population. AB - Mutations in the BRCA1 and BRCA2 genes are known to predispose to breast cancer. In Finland, however, only 21% of all breast cancer families have mutations in these genes. Recent studies have shown that large genomic alterations of BRCA1 are common in many countries. Because such alterations will be missed in conventional mutation screening strategies, we decided to screen Finnish breast and ovarian cancer families for genomic alterations by using a multiplex polymerase chain reaction method. The most characteristic features of BRCA1 related breast cancer were used to select patients, namely (1) both breast and ovarian cancer in the family (48 patients), (2) four or more breast cancers in family (22 patients), or (3) young age (< or =40 years) of onset (58 patients). A total of 128 patients were included in the study. All exons of BRCA1 were analyzed but no alterations were found. This study excludes the frequent occurrence of large genomic alterations in the BRCA1 gene in Finland. Here, again, Finland differs from other countries with a mixed population structure. Our results are in agreement with the common hypothesis that there are still unknown breast cancer susceptibility gene(s) that are responsible for breast cancer predisposition. PMID- 16271957 TI - CD79a expression in acute myeloid leukemia t(8;21) and the importance of cytogenetics in the diagnosis of leukemias with immunophenotypic ambiguity. AB - Acute leukemias that express antigens associated with more than one lineage have been classified as acute lymphocytic leukemia with myeloid markers, acute myeloid leukemia with lymphoid markers, or biphenotypic acute leukemia (BAL). Antibody to cytoplasmic CD79a has been recently introduced to flow cytometry. CD79a functions in and has a high degree of specificity for B-cell differentiation. It has only recently begun to be reported in biphenotypic acute leukemias. Cases of acute leukemia submitted to the flow cytometry laboratory were retrospectively reviewed beginning from the time analysis for cytoplasmic CD79a was added to leukemia and lymphoma panels. Among 89 cases of AML, 2 showed strong coexpression of CD79a. Both cases were differentiated FAB AML-M2 and demonstrated the t(8;21) with cytogenetics and the AML1/ETO rearrangement with fluorescence in situ hybridization (FISH). These are recurring abnormalities in FAB AML-M2. The immunophenotyping met proposed scoring criteria for a diagnosis of BAL. Nevertheless, the cytogenetic and FISH findings indicate that CD79a, despite its specificity for B-cell differentiation, represented the aberrant presence of a B cell antigen in leukemias of distinct myeloid linage. It is doubtful that, in this setting, CD79a expression should be considered a manifestation of lineage ambiguity. PMID- 16271958 TI - Expression patterns of the LPP-HMGA2 fusion transcript in pulmonary chondroid hamartomas with t(3;12)(q27 approximately 28;q14 approximately 15). AB - The high frequency of the t(3;12)(q27 approximately 28; q14 approximately 15) in lipomas and pulmonary chondroid hamartomas (PCHs) makes the HMGA2-LPP fusion gene the most frequent fusion gene in human tumors. We analyzed 11 PCHs with a t(3;12)(q27 approximately 28;q14 approximately 15) for the expression of the LPP HMGA2 fusion transcript. In a previous study, all of these tumors were shown to express the HMGA2-LPP fusion transcript, composed of exons 1-3 of HMGA2 and exons 9-11 of LPP. In the present study, reverse transcriptase-polymerase chain reaction revealed the expression of the reciprocal fusion transcript in 8 of 11 cases. In all positive tumors, the reciprocal fusion transcripts had the same structure, namely, exons 1-8 of LPP and exons 4-5 of HMGA2 encoding a protein composed of the proline-rich region and the first LIM-domain of LPP and the acidic tail of HMGA2. To our knowledge, this is the first report of the expression of the LPP-HMGA2 fusion transcript in a series of PCHs. Its frequent occurrence in PCHs indicates the absence of a larger deletion of the LPP locus accompanying the translocation, such as has been described in a lipoma. Thus, based on this one finding, a role of LPP-HMGA2 in PCH should be considered. PMID- 16271959 TI - Translocation (6;17)(q23;q11.2): a novel cytogenetic abnormality in congenital acute myeloid leukemia? AB - Congenital leukemia occurring within 4 weeks of birth is extremely rare and, excluding transient neonatal myeloproliferation associated with Down syndrome, makes up approximately 1% of childhood leukemias. It is usually seen as acute myeloid leukemia (AML), most frequently French-American-British (FAB) types M4 and M5. Recurrent cytogenetic abnormalities have been reported in this group, and in approximately one third of cases the MLL gene at 11q23 is involved. These patients generally have a poor prognosis. We present a case of congenital leukemia (AML FAB type M1) with an acquired translocation between chromosomes 6 and 17. PMID- 16271960 TI - Poor clinical course in a child with myelodysplastic syndrome and del(13)(q14q22). AB - Myelodysplastic syndromes (MDS) are rare in children, representing 3% or less of all hematopoietic malignancies. Cytogenetic abnormalities, such as -7/7q-, +8, and +21 have been reported in 55-80% of children with MDS. Cytogenetic studies have an important impact on diagnosis, treatment selection, and monitoring therapeutic protocols when combined with morphologic data. We report on a pediatric case of MDS with the presence of the rare clonal abnormality del(13)(q14q22) which underwent a malignant transformation to leukemia and ran a very poor clinical course. PMID- 16271961 TI - Rarity of IgH translocations in Waldenstrom macroglobulinemia. AB - Comparatively little is known of the cytogenetics of Waldenstrom macroglobulinemia (WM). This is primarily due to the low proliferation of the clonal B cells, which precludes conventional karyotyping in many cases. Translocations involving the immunoglobulin heavy chain (IGH) gene at 14q32 are characteristic of many B-cell lymphomas and myelomas. Initial reports suggested that the t(9;14) was characteristic of lymphoplasmacytic lymphoma (the underlying pathological diagnosis in WM), but subsequent studies have failed to confirm the uniqueness of the translocation. To clarify this, we examined 69 cases of WM with interphase fluorescence in situ hybridization and failed to demonstrate an IgH translocation in 67 (97%). We conclude that IGH translocations are not a feature of WM, and the implications of this finding are discussed. PMID- 16271963 TI - A cryptic three-way translocation involving chromosomes 8, 14, and 21 in a case of acute myeloid leukemia subtype M1. PMID- 16271962 TI - Renal oncocytoma with loss of chromosomes Y and 1 evolving to papillary carcinoma in connection with gain of chromosome 7. Coincidence or progression? AB - Hybrid tumors of the kidney are not rare. Previous studies of hybrid renal tumors have been valuable for the understanding of the pathogenesis and progression pathways of renal cell neoplasm. In this paper we describe the morphologic, immunohistochemical, and genetic features of 2 oncocytomas with evolving papillary renal cell carcinoma (PRCC) in a nephrectomy specimen of a 60-year old male. The patient was referred for urologic oncology consultation after the incidental discovery of a renal tumor. Nephrectomy was performed and two separate masses were present grossly. The tumors were stained with hematoxylin and eosin, cytokeratin 7 and vimentin. Genetic studies included conventional metaphase cytogenetics and fluorescence in situ hybridization (FISH). Morphologically, both tumors were oncocytomas with numerous microscopic papillary nests and psammoma bodies. Papillary carcinoma nests were highlighted with cytokeratin 7 and vimentin positivity and were more prominent in the larger tumor. Conventional cytogenetics and FISH demonstrated loss of chromosomes Y and 1 and gains of chromosome 7. We postulate that the PRCC represents a neoplastic progression by the gain of chromosome 7 oncocytoma with -Y and -1. PMID- 16271964 TI - Absence of mutations in DICE1/DDX26 gene in human cancer cell lines with frequent 13q14 deletions. PMID- 16271965 TI - Identification of a complex (11;17;15) translocation in acute promyelocytic leukemia. PMID- 16271966 TI - Reconsidering cell line cross-contamination in NCOL-1. PMID- 16271969 TI - Montane speciation patterns in Ithomiola butterflies (Lepidoptera: Riodinidae): are they consistently moving up in the world? AB - Tropical lowland areas have often been seen as the centres of terrestrial species proliferation, but recent evidence suggests that young species may be more frequent in montane areas. Several montane speciation modes have been proposed, but their relative frequencies and predominant evolutionary sequence remain unclear because so few biogeographic and phylogenetic studies have tested such questions. I use morphological data to generate a phylogenetic hypothesis for all 11 species of the riodinid butterfly genus Ithomiola (Riodininae: Mesosemiini: Napaeina). These species are shown here to be all strictly geographically and elevationally allo- or parapatrically distributed with respect to their closest relatives in lowland and montane regions throughout the Neotropics. The overwhelming pattern in Ithomiola is of repeated upward parapatric speciation across an elevational gradient, and the genus appears to provide the clearest example to date of vertical montane speciation. All of the young derived species are montane and all of the old basal species are confined to the lowlands, supporting the hypothesis of montane regions largely as 'species pumps' and lowland regions as 'museums'. Possible reasons for the post-speciation maintenance of parapatric ranges in Ithomiola are discussed. PMID- 16271968 TI - A candidate locus for variation in dispersal rate in a butterfly metapopulation. AB - Frequent extinctions of local populations in metapopulations create opportunities for migrant females to establish new populations. In a metapopulation of the Glanville fritillary butterfly (Melitaea cinxia), more mobile individuals are more likely to establish new populations, especially in habitat patches that are poorly connected to existing populations. Here we show that flight metabolic rate and the frequency of a specific allele of the metabolic enzyme phosphoglucose isomerase (pgi) were both highest in newly established, isolated populations. Furthermore, genotypes with this pgi allele had elevated flight metabolic rates. These results suggest that genetic variation in pgi or a closely linked locus has a direct effect on flight metabolism, dispersal rate, and thereby on metapopulation dynamics in this species. These results also contribute to an emerging understanding of the mechanisms by which population turnover in heterogeneous landscapes may maintain genetic and phenotypic variation across populations. PMID- 16271970 TI - Sex and segregation in temperate bats. AB - Many temperate insectivorous bats show marked sexual segregation during the summer, but in spectacular, pre-hibernation swarming, gather at caves to mate. In many species, sexual segregation is probably due to a gradient in aerial insect availability that confines females to lower elevations, where high reproductive costs are met by an abundant and reliable food supply. In the hawking and trawling Myotis daubentonii, we show that alongside inter-sexual segregation, there is intra-male segregation and suggest that this results from the exclusion of most males from high-quality habitat. These apparently excluded males suffer reduced foraging efficiency and mating success relative to males that roost with the females in summer. Changes in resources and behaviour at the end of the summer lead to a change in strategy that gives all males a chance to mate during swarming, but this does not overcome the paternity advantage to males that spend the summer with the females. PMID- 16271972 TI - Helpers benefit offspring in both the short and long-term in the cooperatively breeding banded mongoose. AB - Helpers in cooperative and communal breeding species are thought to accrue fitness benefits through improving the condition and survival of the offspring that they care for, yet few studies have shown conclusively that helpers benefit the offspring they rear. Using a novel approach to control for potentially confounding group-specific variables, I compare banded mongoose (Mungos mungo) offspring within the same litter that differ in the amount of time they spend with a helper, and hence the amount of care they receive. I show that pups that spend more time in close proximity to a helper are fed more, grow faster and have a higher probability of survival to independence than their littermates. Moreover, high growth rates during development reduce the age at which females breed for the first time, suggesting that helpers can improve the future fecundity of the offspring for which they care. These results provide strong evidence that it is the amount of investment per se that benefits offspring, rather than some correlate such as territory quality, and validate the assumption that helpers improve the reproductive success of breeders, and hence may gain fitness benefits from their actions. Furthermore, the finding that helpers may benefit offspring in the long-term suggests that current studies underestimate the fitness benefits that helpers gain from rearing the offspring of others. PMID- 16271971 TI - Females use self-referent cues to avoid mating with previous mates. AB - Females of many species mate repeatedly throughout their lives, often with many different males (polyandry). Females can secure genetic benefits by maximizing their diversity of mating partners, and might be expected, therefore, to forego matings with previous partners in favour of novel males. Indeed, a female preference for novel mating partners has been shown in several taxa, but the mechanism by which females distinguish between novel males and previous mates remains unknown. We show that female crickets (Gryllodes sigillatus) mark males with their own unique chemical signatures during mating, enabling females to recognize prior mates in subsequent encounters and to avoid remating with them. Because self-referent chemosensory cues provide females with a simple, but reliable mechanism of identifying individuals with whom they have mated without requiring any special cognitive ability, they may be a widespread means by which females across a broad range of animal mating systems maximize the genetic benefits of polyandry. PMID- 16271973 TI - Adaptive male effects on female ageing in seed beetles. AB - Selection can favour the evolution of a high reproductive rate early in life even when this results in a subsequent increase in the rate of mortality, because selection is relatively weak late in life. However, the optimal reproductive schedule of a female may be suboptimal to any one of her mates, and males may thus be selected to modulate female reproductive rate. Owing to such sexual conflict, coevolution between males and females may contribute to the evolution of senescence. By using replicated beetle populations selected for reproduction at an early or late age, we show that males evolve to affect senescence in females in a manner consistent with the genetic interests of males. 'Late' males evolved to decelerate senescence and increase the lifespan of control females, relative to 'early' males. Our findings demonstrate that adaptive evolution in one sex may involve its effects on senescence in the other, showing that the evolution of optimal life histories in one sex may be either facilitated or constrained by genes expressed in the other. PMID- 16271974 TI - Patterns of male sterility in a grasshopper hybrid zone imply accumulation of hybrid incompatibilities without selection. AB - It is now widely accepted that post-zygotic reproductive isolation is the result of negative epistatic interactions between derived alleles fixed independently at different loci in diverging populations (the Dobzhansky-Muller model). What is less clear is the nature of the loci involved and whether the derived alleles increase in frequency through genetic drift, or as a result of natural or sexual selection. If incompatible alleles are fixed by selection, transient polymorphisms will be rare and clines for these alleles will be steep where divergent populations meet. If they evolve by drift, populations are expected to harbour substantial genetic variation in compatibility and alleles will introgress across hybrid zones once they recombine onto a genetic background with which they are compatible. Here we show that variation in male sterility in a naturally occurring Chorthippus parallelus grasshopper hybrid zone conforms to the neutral expectations. Asymmetrical clines for male sterility have long tails of introgression and populations distant from the zone centre show significant genetic variation for compatibility. Our data contrast with recent observations on 'speciation genes' that have diverged as a result of strong natural selection. PMID- 16271975 TI - The pathogen causing Dutch elm disease makes host trees attract insect vectors. AB - Dutch elm disease is caused by the fungal pathogen Ophiostoma novo-ulmi which is transmitted by the native elm bark beetle, Hylurgopinus rufipes. We have found that four semiochemicals (the monoterpene (-)-beta-pinene and the sesquiterpenes (-)-alpha-cubebene, (+)-spiroaxa-5,7-diene and (+)-delta-cadinene) from diseased American elms, Ulmus americana, synergistically attract H. rufipes, and that sesquiterpene emission is upregulated in elm trees inoculated with O. novo-ulmi. The fungus thus manipulates host trees to enhance their apparency to foraging beetles, a strategy that increases the probability of transportation of the pathogen to new hosts. PMID- 16271977 TI - Evolutionary implications of the adaptation to different immune systems in a parasite with a complex life cycle. AB - Many diseases are caused by parasites with complex life cycles that involve several hosts. If parasites cope better with only one of the different types of immune systems of their host species, we might expect a trade-off in parasite performance in the different hosts, that likely influences the evolution of virulence. We tested this hypothesis in a naturally co-evolving host-parasite system consisting of the tapeworm Schistocephalus solidus and its intermediate hosts, a copepod, Macrocyclops albidus, and the three-spined stickleback Gasterosteus aculeatus. We did not find a trade-off between infection success in the two hosts. Rather, tapeworms seem to trade-off adaptation towards different parts of their hosts' immune systems. Worm sibships that performed better in the invertebrate host also seem to be able to evade detection by the fish innate defence systems, i.e. induce lower levels of activation of innate immune components. These worm variants were less harmful for the fish host likely due to reduced costs of an activated innate immune system. These findings substantiate the impact of both hosts' immune systems on parasite performance and virulence. PMID- 16271978 TI - A within-species warning function for an aposematic signal. AB - Aposematic, or warning, signals are generally interspecific in form: one species advertises noxiousness to a predator or parasite species. In a study of the pipevine swallowtail butterfly (Battus philenor), we show that a pattern of colouration in the caterpillars that is considered to be aposematic in the context of attack by natural enemies also deters oviposition by conspecific females. In field and laboratory assays, females avoided oviposition on plants bearing live conspecific larvae. Females avoided oviposition on plants bearing artificially constructed models identical to larvae in shape, size and colour pattern. Finally, oviposition on plants harbouring a model bearing the larval colour pattern was reduced relative to plants bearing a leaf-green model, suggesting that the larval colour pattern was essential for avoidance. We discuss how intraspecific and interspecific processes might interact in the evolution of an aposematic signal. PMID- 16271976 TI - Conflict between parasites with different transmission strategies infecting an amphipod host. AB - Competition between parasites within a host can influence the evolution of parasite virulence and host resistance, but few studies examine the effects of unrelated parasites with conflicting transmission strategies infecting the same host. Vertically transmitted (VT) parasites, transmitted from mother to offspring, are in conflict with virulent, horizontally transmitted (HT) parasites, because healthy hosts are necessary to maximize VT parasite fitness. Resolution of the conflict between these parasites should lead to the evolution of one of two strategies: avoidance, or sabotage of HT parasite virulence by the VT parasite. We investigated two co-infecting parasites in the amphipod host, Gammarus roeseli: VT microsporidia have little effect on host fitness, but acanthocephala modify host behaviour, increasing the probability that the amphipod is predated by the acanthocephalan's definitive host. We found evidence for sabotage: the behavioural manipulation induced by the Acanthocephala Polymorphus minutus was weaker in hosts also infected by the microsporidia Dictyocoela sp. (roeselum) compared to hosts infected by P. minutus alone. Such conflicts may explain a significant portion of the variation generally observed in behavioural measures, and since VT parasites are ubiquitous in invertebrates, often passing undetected, conflict via transmission may be of great importance in the study of host-parasite relationships. PMID- 16271980 TI - Not so ancient: the extant crown group of Nothofagus represents a post-Gondwanan radiation. AB - This study uses a molecular-dating approach to test hypotheses about the biogeography of Nothofagus. The molecular modelling suggests that the present-day subgenera and species date from a radiation that most likely commenced between 55 and 40 Myr ago. This rules out the possibility of a reconciled all-vicariance hypothesis for the biogeography of extant Nothofagus. However, the molecular dates for divergences between Australasian and South American taxa are consistent with the rifting of Australia and South America from Antarctica. The molecular dates further suggest a dispersal of subgenera Lophozonia and Fuscospora between Australia and New Zealand after the onset of the Antarctic Circumpolar Current and west wind drift. It appears likely that the New Caledonian lineage of subgenus Brassospora diverged from the New Guinean lineage elsewhere, prior to colonizing New Caledonia. The molecular approach strongly supports fossil-based estimates that Nothofagus diverged from the rest of Fagales more than 84 Myr ago. However, the mid-Cenozoic estimate for the diversification of the four extant subgenera conflicts with the palynological interpretation because pollen fossils, attributed to all four extant subgenera, were widespread across the Weddellian province of Gondwana about 71 Myr ago. The discrepancy between the pollen and molecular dates exists even when confidence intervals from several sources of error are taken into account. In contrast, the molecular age estimates are consistent with macrofossil dates. The incongruence between pollen fossils and molecular dates could be resolved if the early pollen types represent extinct lineages, with similar types later evolving independently in the extant lineages. PMID- 16271979 TI - Strikingly variable divergence times inferred across an Amazonian butterfly 'suture zone'. AB - 'Suture zones' are areas where hybrid and contact zones of multiple taxa are clustered. Such zones have been regarded as strong evidence for allopatric divergence by proponents of the Pleistocene forest refugia theory, a vicariance hypothesis frequently used to explain diversification in the Amazon basin. A central prediction of the refugia and other vicariance theories is that the taxa should have a common history so that divergence times should be coincident among taxa. A suture zone for Ithomiinae butterflies near Tarapoto, NE Peru, was therefore studied to examine divergence times of taxa in contact across the zone. We sequenced 1619bp of the mitochondrial COI/COII region in 172 individuals of 31 species from across the suture zone. Inferred divergence times differed remarkably, with divergence between some pairs of widespread species (each of which may have two or more subspecies interacting in the zone, as in the genus Melinaea) being considerably less than that between hybridizing subspecies in other genera (for instance in Oleria). Our data therefore strongly refute a simple hypothesis of simultaneous vicariance and suggest that ongoing parapatric or other modes of differentiation in continuous forest may be important in driving diversification in Amazonia. PMID- 16271981 TI - From Europe to America: pliocene to recent trans-atlantic expansion of cold-water north atlantic molluscs. AB - Data on the geographical distribution, phylogeny and fossil record of cool temperate North Atlantic shell-bearing molluscs that live in waters shallower than 100 m depth belong to two biogeographic provinces, one in eastern North America north of Cape Cod, the other in northern Europe. Amphi-Atlantic species, which are found in both provinces, comprise 30.8% of the 402 species in the northeastern Atlantic and 47.3% of the 262 species in the northwestern Atlantic. Some 54.8% of these amphi-Atlantic species have phylogenetic origins in the North Pacific. Comparisons among fossil Atlantic faunas show that amphi-Atlantic distributions became established in the Middle Pliocene (about 3.5 million years ago), and that all represent westward expansions of European taxa to North America. No American taxa spread eastward to Europe without human assistance. These results are in accord with previous phylogeographic studies among populations within several amphi-Atlantic species. Explanations for the unidirectional expansion of species across the Atlantic remain uncertain, but may include smaller size and greater prior extinction of the North American as compared to the European fauna and biased transport mechanisms. Destruction of the European source fauna may jeopardize faunas on both sides of the Atlantic. PMID- 16271983 TI - Laboratory diagnosis of iron deficiency anaemia. PMID- 16271982 TI - Non-random nature of spontaneous mIPSCs in mouse auditory brainstem neurons revealed by recurrence quantification analysis. AB - A change in the spontaneous release of neurotransmitter is a useful indicator of processes occurring within presynaptic terminals. Linear techniques (e.g. Fourier transform) have been used to analyse spontaneous synaptic events in previous studies, but such methods are inappropriate if the timing pattern is complex. We have investigated spontaneous glycinergic miniature synaptic currents (mIPSCs) in principal cells of the medial nucleus of the trapezoid body. The random versus deterministic (or periodic) nature of mIPSCs was assessed using recurrence quantification analysis. Nonlinear methods were then used to quantify any detected determinism in spontaneous release, and to test for chaotic or fractal patterns. Modelling demonstrated that this procedure is much more sensitive in detecting periodicities than conventional techniques. mIPSCs were found to exhibit periodicities that were abolished by blockade of internal calcium stores with ryanodine, suggesting calcium oscillations in the presynaptic inhibitory terminals. Analysis indicated that mIPSC occurrences were chaotic in nature. Furthermore, periodicities were less evident in congenitally deaf mice than in normal mice, indicating that appropriate neural activity during development is necessary for the expression of deterministic chaos in mIPSC patterns. We suggest that chaotic oscillations of mIPSC occurrences play a physiological role in signal processing in the auditory brainstem. PMID- 16271984 TI - Molecular diagnosis of adult-type hypolactasia (lactase non-persistence). PMID- 16271985 TI - Serum lipid and apolipoprotein profiles in newborns and six-year-old children: the Tallinn Young Family Study. AB - Seventy children aged 6 years (34 boys, 36 girls) were studied for cardiovascular risk factors. Among the children 40 had also been investigated at birth. The aim of the study was to determine changes in serum lipoprotein parameters from birth up to preschool age and to assess the role of some relevant factors that might affect the process. An obvious association was found between serum apolipoprotein (apo) B levels, the apoB/apoA-I ratio and lipoprotein(a) (Lp(a)) levels at birth and at 6 years of age (r = 0.43; p<0.05, r = 0.73; p<0.0001 and r = 0.81; p<0.0001, respectively). Thirty percent of children who were in the top quartile by apoB or total cholesterol levels and 66.7% of those in this quartile by apoB/apoA-I ratio at birth remained in the top quartiles also in the follow-up study. The significantly higher apoB/apoA-I ratio in newborns and the apoB/apoA-I and apoB values in the 6-year-old children were observed in the carrier apoE4 isoform as compared to E3 homozygotes. A significant influence of apoE polymorphism on serum apoB/apoA-I ratio and apoB level in preschool children was confirmed by ANOVA one-way analysis of variance. In a multiple regression analysis from all the studied factors, the independent determinants of apoB level in preschool age were apoE phenotype, gestational age and Apgar score in the first minute of life. Thus, tracking of serum Lp(a), apoB, apoB/apoA-I ratio and total cholesterol levels from birth up to 6 years of age was demonstrated. The association between apoE polymorphism and serum lipoprotein parameters became more obvious after the first 6 years of life. PMID- 16271986 TI - A method to estimate the uncertainty of measurements in a conglomerate of instruments/laboratories. AB - A comparison of the performance, at two concentrations, of the measurement procedures for 23 common components is presented. For each component the within-, pure between- and total variations have been calculated and an imprecision profile indicated. Thus a pragmatic, readily and generally applicable method is advised to describe the performance of laboratories under repeatability and reproducibility conditions. The calculated total variation is compared with the results estimated from the biological variation of the properties. In most cases the within-laboratory variation is better than that given by the reference database, whereas in about half of the procedures the total variation exceeds that of the reference database. The outcome illustrates the transferability problems that multicentre studies and the diagnosis of patients face by measuring samples using a conglomerate of instruments/laboratories that are not aligned. The calculation procedures are detailed in an appendix and the calculation and presentation of the results are made with a custom-made worksheet program. PMID- 16271988 TI - Usefulness of soluble transferrin receptor and ferritin in iron deficiency and chronic disease. AB - Soluble transferrin receptor (sTfR) is a biochemical parameter used for the detection of iron deficiency in situations where ferritin has limited diagnostic value owing to the present chronic disease. The sTfR concentration was determined in 118 patients divided according to their inflammatory status and underlying disease into groups of patients with iron-deficiency anemia (IDA), anemia of chronic disease (ACD) and patients with a coexisting state of iron deficiency and anemia of chronic disease (ID+ACD). All patients with iron deficiency had elevated sTfR levels, but ferritin concentrations were normal or increased in patients with inflammatory characteristics. Diagnostic efficiencies of sTfR, sTfR/log ferritin index (sTfR/F) and ferritin were evaluated by receiver operating characteristic curve (ROC) analysis. According to the results obtained, the best diagnostic efficiency for differential diagnosis of anemic patients with iron deficiency compared to the control group had a sTfR concentration (0.884) that was significantly higher than ferritin (0.638), but not higher than the calculated ratio sTfR/F (0.820). The cut-off value of the sTfR/F index differentiating the best control group from the IDA and ID+ACD groups was 1.30, and for differentiation of ACD from IDA and ID+ACD, the value was 0.90. Soluble transferrin receptor is an additional parameter to ferritin for the diagnosis of IDA and differential diagnosis of ID+ACD, but calculation of the sTfR/F index did not improve the diagnostic value of determining sTfR alone. PMID- 16271987 TI - Adhesion of T and B lymphocytes to mouse atherosclerotic aortas: association with lesion topology and VCAM-1 expression. AB - OBJECTIVE: Although T and B lymphocytes accumulate in atherosclerotic lesions and play a key role in their growth, the mechanisms involved in the adhesion and recruitment of T and B lymphocytes by the lesions have not been resolved. The aim of this study was to compare T and B lymphocyte adhesion to atherosclerotic arteries and to test the role of VCAM-1 and ICAM-1. MATERIAL AND METHODS: T and B lymphocytes were labelled with red and green fluorescent dyes and incubated with freshly isolated aortas from apolipoprotein-E-deficient mice. In some experiments the aortas were pre-incubated with specific monoclonal antibodies. After washing, the adhering cells were detected by confocal laser scanning microscopy. RESULTS: The number of T and B lymphocytes that adhered to the aortic intimal surface was similar in both lesioned and non-lesioned areas and in the shoulder region of the lesions. However, the adhesion of T and B lymphocytes was significantly higher in the shoulder regions compared with the lesioned (p<0.0001) and non-lesioned areas of the aorta (p<0.0001). After pre-incubation of the aortas with antibodies against VCAM-1 or ICAM-1, the lymphocyte adhesions in lesioned areas were 42 % (p = 0.04) and 55 % (p = 0.17), respectively, of those in lesioned areas that had been pre-incubated with a control antibody. However, although VCAM-1 protein expression was most pronounced in the shoulder region, the lymphocyte adhesions in the shoulder region and in non-lesioned areas were unaffected by pre incubation with VCAM-1 antibodies. CONCLUSIONS: The results suggest that adhesion of T and B lymphocytes to mouse aortic endothelium is similar, is affected by lesion topology and is dependent on VCAM-1 expression over the core of atherosclerotic lesions. PMID- 16271989 TI - Trans-sodium crocetinate does not affect oxygen uptake in rats during treadmill running. AB - Trans-sodium crocetinate (TSC), the isomer of the carotenoid compound crocetin, is found markedly to increase survival in hemorrhagic shock subsequent to 50-60% blood loss, mainly via restored resting oxygen consumption (VO(2)), blood pressure and heart rate. The proposed mechanism is that TSC increases oxygen diffusivity, and thus availability, in plasma. If this were found to be a prominent feature in the oxygen transfer from blood to skeletal muscle fiber mitochondria, increased VO(2) during exercise would be expected because of reduced partial pressure of venous oxygen (increased utilization), which we aimed to elucidate in this study. Male Sprague-Dawley rats were intravenously injected with 0.3 mL kg(-1) TSC (40 microg mL(-1)) or placebo and immediately thereafter tested on a ramped treadmill test protocol. Rats were introduced to the experimental protocols beforehand. Administration of TSC had a neutral effect on submaximal and maximal VO(2) (VO(2max)) as well as running performance measured as maximal running time and maximal aerobic running velocity. Thus, in this study we cannot report any effects of TSC on steady-state submaximal VO(2) or VO(2max) at exhaustive exercise. PMID- 16271990 TI - Hyperproinsulinemia segregates young adult patients with newly diagnosed autoimmune (type 1) and non-autoimmune (type 2) diabetes. AB - OBJECTIVE: To investigate whether measurements of proinsulin and/or intermediate proinsulin degradation products could be used to differentiate between autoimmune (type 1) and non-autoimmune (type 2) diabetes in young adults. MATERIAL AND METHODS: Total proinsulin, intact proinsulin and 32,33 split proinsulin concentrations were measured in 25 patients aged 15-34 years with type 1 diabetes, as defined by the presence of at least two positive islet autoantibodies, and in 23 antibody-negative patients of similar age with type 2 diabetes, at the time of clinical onset of diabetes and at 3-4 months thereafter. Comparisons were made with data from 25 healthy subjects matched for gender and age. RESULTS: Plasma levels of total proinsulin, intact proinsulin and 32,33 split proinsulin were significantly increased 2-3-fold in the patients with newly diagnosed type 2 diabetes as compared with the controls, both in absolute terms (p<0.0001) and when related to circulating insulin (p<0.01-0.0002). In contrast, absolute proinsulin and 32,33 split proinsulin concentrations were significantly lower in patients with onset of type 1 diabetes than in controls. When proinsulin and split proinsulin release were related to plasma insulin, however, similar ratios were found in the type 1 diabetes patients and in controls. Using the 90th percentile for total proinsulin in the control group as the cut-off, the sensitivity and specificity for differentiation between autoimmune and non autoimmune diabetes were 87% and 92%, respectively. At 3-4 months after clinical onset of diabetes, proinsulin secretion was still 2-3 times higher in type 2 than in type 1 diabetes patients (p<0.001). CONCLUSIONS: Young adult patients with newly diagnosed type 2 diabetes display disproportionate hyperproinsulinemia, whereas proinsulin secretion appears to be normal in patients with clinical onset of type 1 diabetes. Evaluation of proinsulin and 32,33 split proinsulin concentrations may be useful as a diagnostic tool in differentiating between autoimmune and non-autoimmune diabetes in young adults, particularly in those lacking islet autoantibodies at diagnosis. PMID- 16271991 TI - Polymorphism in the coding part of the sterol 12alpha-hydroxylase gene does not explain the marked differences in the ratio of cholic acid and chenodeoxycholic acid in human bile. AB - OBJECTIVE: In humans, two primary bile acids are synthesized: cholic acid (CA) and chenodeoxycholic acid (CDCA), the first and rate-limiting enzyme being cholesterol 7alpha-hydroxylase (CYP7A1). CA has one more hydroxyl group at position 12alpha. This hydroxylation is carried out by the sterol 12alpha hydroxylase (CYP8B1). Earlier, we and others have noticed a marked variation in the ratio between CA and CDCA in human bile. The aim of this study was to investigate whether this marked difference could be due to a genetic polymorphism in the gene of the CYP8B1. MATERIAL AND METHODS: Screening for genetic polymorphisms was carried out in a 2.4-kb-long area including the exon and part of the promoter region in subjects who had undergone cholecystectomy earlier, and where bile acid analysis had been performed. Among these subjects those with very high or low CA/CDCA ratios (ranging from 0.9 to 6.8) were investigated. The subjects were all female, normolipidaemic, having normal weight and a normal thyroid function. RESULTS: No polymorphisms were found in the investigated sequence. However, a statistically significant correlation was found between the activity of the CYP7A1 and the ratio between CA and CDCA. The difference in ratio could, at least in part, be explained by the difference in rate of bile acid synthesis. CONCLUSION: The difference in ratio between CA and CDCA cannot be explained by a polymorphism in the coding area of the CYP8B1. PMID- 16271992 TI - Plasma antibody titres to heat shock proteins-60, -65 and-70: their relationship to coronary risk factors in dyslipidaemic patients and healthy individuals. AB - OBJECTIVE: To investigate the factors that may affect antibody titres to heat shock proteins (Hsp)-60, -65 and -70, and serum C-reactive protein (CRP) concentrations in patients with dyslipidaemia and other features of the metabolic syndrome as defined by ATPIII criteria. MATERIAL AND METHODS: The study comprised 237 dyslipidaemia patients and 135 healthy individuals recruited from amongst university and hospital employees. RESULTS: Compared to the healthy individuals, the dyslipidaemic patients had higher antibody titres to Hsp-60 (p<0.01), Hsp-65 (p<0.001) and Hsp-70 (p<0.05), and higher serum CRP concentrations (p<0.001). The best-fitting multifactorial models revealed that known coronary risk factors explained little of the variation in Hsp antibody titres: 3 % for Hsp-60, 1 % for Hsp-65 and 4 % for Hsp-70 amongst the dyslipidaemic subjects. The corresponding values for the subgroup with the metabolic syndrome were 8 %, 3 % and 1 %, respectively. In contrast, the best-fitting model explained 13.5 % of the variation in serum CRP concentrations among the dyslipidaemic patients, obesity being a major determinant; and 14 % in the subgroup with metabolic syndrome. CONCLUSIONS: The higher antibody titres to Hsp-60, -65, and -70 in the dyslipidaemic patients may be related to a heightened state of immunoactivation associated with atherosclerosis in this group. Our data indicate that antibody titres to these Hsps are not associated with the classical coronary risk factors, although serum high sensitivity (hs)CRP concentrations were significantly related to obesity. PMID- 16271994 TI - Mode of classification of source material as citable items skews journal impact factor calculations. PMID- 16271993 TI - Validation of a self-administered modified CAGE test (CAGE-C) in a somatic hospital ward: comparison with biochemical markers. AB - OBJECTIVE: The time frame for the original CAGE questionnaire is lifetime and it does not quantify drinking frequency and may be less suitable in a population with very few teetotalers. The purpose of this study was to validate a variant of the CAGE questionnaire and compare it with the outcome of a thorough interview according to DSM-III and ICD-10 criteria and to the outcome of biochemical markers in inpatients in a somatic hospital setting. MATERIAL AND METHODS: The questionnaire and biochemical markers were tested on a random sample of 130 patients admitted to a department of orthopedic surgery. The result of a diagnostic interview with a trained staff member from the local alcohol treatment unit was used as the gold standard. Data were analyzed by means of receiver operating characteristic (ROC) curves. RESULTS: In this population 25 % had an alcohol problem and the questionnaire proved to be valid, with a sensitivity and specificity of 0.94 and 0.88, respectively, while the positive predictive value (PVpos) was 0.73 and the negative predictive value (PVneg) was 0.98. Carbohydrate deficient transferrin (CDT) had a sensitivity and a specificity of 0.47 and 0.96, and PVpos and PVneg of 0.80 and 0.85, respectively. CONCLUSIONS: This new diagnostic questionnaire is simple, easy to administer and suitable for screening purposes in populations with a high prevalence of at-risk drinkers. PMID- 16271995 TI - Iron status among 3005 women aged 20-55 years in central Norway: the Nord Trondelag Health Study (the HUNT Study). PMID- 16272000 TI - The need for self-report data in the assessment of stuttering therapy efficacy: repetitions and prolongations of speech. The stuttering syndrome. AB - BACKGROUND: Bloodstein reviewed hundreds of studies that investigated the efficacy of therapeutic protocols for ameliorating the stuttering syndrome. Surprisingly, almost all were effective in significantly reducing overtly perceptible behaviours such as repetitions and prolongations of speech sounds. These results seem highly improbable considering that many of the treatment methods were diametrically opposed in their principles and implementation procedures (e.g. psychoanalysis, drug therapy, behaviourism, cognitive behavioural therapy and auditory feedback devices with rate control, etc.). In addition, time and more ecologically valid methods such as self-report measures demonstrate that overt measures of success are tenuous, their ameliorative effects tend to diminish drastically over time and show poor generalizability. Further, the real conundrum in stuttering therapy is the failure to acknowledge stuttering as a complete syndrome of continuous compensatory behaviours. AIMS: To highlight how self-report measures serve as a primary tool to understand the syndrome-like nature of stuttering and to test the efficacy of the therapy outside the confines of the clinic and the needs of the people who stutter. METHODS & PROCEDURES/OUTCOMES & RESULTS: In the past, therapeutic efficacy has typically been measured by the reduction in overtly observable and countable events of stuttering such as repetitions and prolongations. However, recent neuroimaging data and our research suggest that the stuttering syndrome is more than the mere presence of peripheral speech disruptions. Stuttering is a central, experiential sense of 'loss of control' that manifests itself across a continuum of compensatory behaviours from the central nervous system outwards to the speech periphery. In other words, aberrant neural activity, as well as covert stuttering behaviours, subperceptual stuttering forms and overt speech disruptions are all effects or compensations for the central involuntary 'neural block'. Hence, by counting only perceptible portions of the disorder, efficacy measures 'fail to capture' the experiential sense of 'loss of control' and the covert compensatory behaviours of the disorder (i.e. avoidances of words or situations, substitutions, circumlocutions, subperceptual stuttering forms, etc.). Furthermore, unnatural sounding speech, decreased ease of speech production, elevated levels of clinic room fluency and poor reliability in counting stuttering behaviours confound the overt measures in the clinic milieu. Therefore, while overt measures remain important, used in isolation, they cannot provide a 'true metric' of efficacy. CONCLUSIONS: Any efficient and effective means of evaluating intervention methods over the long-term should include a form of self-report as a primary tool as it best accesses the experiential sense of 'loss of control' and other covert behaviours. Overt measures should be used to supplement or complement the self-report data. PMID- 16272001 TI - Resistance of grammatical impairment to computerized comprehension training in children with specific and non-specific language impairments. AB - BACKGROUND: Receptive language impairments in school-age children have a poor prognosis, yet there is a dearth of research on effective interventions. AIMS: Children's responses to a computerized grammatical training program were evaluated to consider whether repeated responding to spoken sentences with variable semantic content and the same syntactic structure would lead to consistent and fluent comprehension. METHODS & PROCEDURES: Children with receptive language impairments aged from 8 to 13 years were randomly assigned to three groups: Group S (n = 12) responded to reversible sentences in a computerized game, using speech stimuli with pauses before critical phrases. Group M (n = 12) had the same stimuli acoustically modified to lengthen and amplify dynamic portions of the signal. Group U (n = 9) was an untrained control group. On average, children in groups S and M completed over 1000 training trials, focusing on training comprehension of reversible sentences. OUTCOMES & RESULTS: Although responses speeded up over the course of training, and most children performed well above chance, accuracy typically remained below 95% correct for constructions such as above/below and reversible active/passive. Trained groups did not differ from untrained children on language or auditory outcomes. There was no evidence that acoustically modified speech input enhanced comprehension. CONCLUSIONS: Rote training of comprehension of reversible sentences does not seem to be an effective approach to remediating such problems. For most children, the pattern of performance suggested that the problem was not a lack of syntactic knowledge, bur rather limited processing capacity that led to failures of on-line computation of meaning. PMID- 16272002 TI - Exploring the effects of communication intervention for developmental pragmatic language impairments: a signal-generation study. AB - BACKGROUND: The remediation of pragmatic problems forms a significant part of the caseload for professionals working with children with communication problems. There is little systematic evidence that demonstrates the benefits of speech and language therapy for children whose difficulties lie primarily within the pragmatic domain or which indicates whether changes in pragmatic behaviours, which are a result of a specific intervention, can be measured over time. AIMS: To generate a signal of change in pragmatic and other language behaviours for children with pragmatic language impairments; to gauge the magnitude and nature of the signal and to make recommendations for future studies. METHODS AND PROCEDURES: A case series of six children with pragmatic language impairments without diagnosis of autism received 8 weeks of individual intensive speech and language therapy supported in a mainstream educational setting in the UK. Measures of pragmatic behaviours in conversation were made at seven data points before and after therapy using Bishop's ALICC procedure. Conversation coders were blind to the point of assessment. Inferential comprehension, narrative, sentence formulation and sentence recall skills were also tested before and after therapy. The opinions of teachers and parents were sought regarding any change in communication and social abilities of the children over time. OUTCOMES AND RESULTS: All children showed change in communication behaviour on some conversational measures, even if the child functioned at the ceiling on standardized language testing. Some conversation measures had more utility as outcome measures than others. Most children showed substantial change on standardized language measures, but there are limitations on the use of these due to heterogeneity within the group. Overall, the intervention produced a signal for change in pragmatics and/or language behaviour in all children. Parent/teacher opinion reported demonstrable change in communication behaviour and engagement in the curriculum. CONCLUSIONS: There is a strong signal that change in pragmatic language behaviour can be measured in well-controlled intervention studies but this signal is complex. Outcome measures should take into account changes in language processing skills that are significantly impaired in many children with PLI. For those children within the PLI group who function at ceiling on language tests, conversational measures may have the potential to signal change, but this finding has not been subjected to group study or to testing in generalized settings. Qualitative data regarding behaviour, classroom engagement and generalization of language gains will be an essential supplement to measuring progress in a diverse population. PMID- 16272003 TI - Early Years Centres for pre-school children with primary language difficulties: what do they cost, and are they cost-effective? AB - BACKGROUND: High levels of early language difficulties raise practical issues about the efficient and effective means of meeting children's needs. Persistent language difficulties place significant financial pressures on health and education services. This has led to large investment in intervention in the early years; yet, little is known about the actual and relative costs of early years provision. AIMS: To profile the different costs incurred by two Early Years Centres (EYCs) partially funded by the charity I CAN and children receiving what might be termed 'routine' NHS speech therapy to provide an analysis of cost efficiency and equity. METHODS & PROCEDURES: Costings for service provision for 91 children (mean age 2;9) were collected. The activity of staff at each site and the cost of staff allocated to services were computed. Data on other resources were also collected. OUTCOMES & RESULTS: The cost per child per session was on average 12 pounds. Despite the longer course of intervention in the first centre (10 compared with 6 weeks), the cost of the course per child was of the same order (245 pounds compared with 253 pounds). The annual cost of the early years provision per child was higher relative to the costs of the NHS provision, 645 pounds compared with 181 pounds in one EYC (A) and 462 pounds compared with 173 pounds in the other (B). When the cost of standard nursery provision was factored in, the difference in annual costs was rather less, with 5298 pounds for the early years provision (EYC A) relative to 4276 pounds in the comparison group. By contrast, the annual cost of early provision rises to 5926 pounds relative to 8861 pounds in the comparison group (EYC B). CONCLUSIONS: The cost of the EYCs is relatively low and given the positive outcomes reported in the study of which this economic evaluation is a part, there is a good case for saying that they represent an efficient use of resources. The strengths and limitations of the economic evaluation are considered and the need for long-term evaluations is highlighted. PMID- 16272004 TI - Perceptions of speech and language therapy amongst UK school and college students: implications for recruitment. AB - BACKGROUND: Communication disorders affect both sexes and people from all ethnic groups, but members of minority ethnic groups and males in the UK are underrepresented in the speech and language therapy profession. Research in the area of recruitment is limited, but a possible explanation is poor awareness and understanding of speech and language therapy as a profession. AIMS: To investigate factors influencing attitudes to a career in speech and language therapy amongst UK school and college students focusing on the similarities and differences between males and females, and between minority ethnic and white students. METHODS & PROCEDURES: A total of 651 male and female school and college students from a range of ethnic groups and all close to selecting degree courses completed a questionnaire designed to examine the attitudes and awareness of speech and language therapy. Eleven semi-structured follow-up interviews were conducted to help understand the questionnaire findings. Quantitative and qualitative analyses examined differences in attitudes and the awareness of speech and language therapy amongst these groups. OUTCOMES & RESULTS: Overall, one-third of participants said they knew nothing about speech and language therapy, and males were significantly less familiar with it. Less than half the participants were aware that speech and language therapy is a degree course, and minority ethnic participants were significantly less likely to know this. Compared with males, females were almost five times as likely to say they would consider a career in speech and language therapy. Participants with relatives in health-related jobs were significantly more likely to consider speech and language therapy than those without such relatives. Compared with white participants, minority ethnic participants said they placed greater importance on studying for a degree, a profession and a scientific career, and were more influenced by a career's prestige and a high salary. CONCLUSIONS: In order to increase the ethnic and gender diversity of speech and language therapists, the profile of the profession needs to be raised with increased awareness of the degree level courses, the scientific, evidence-based nature of the work, and current salary scales. PMID- 16272005 TI - Systematic reviews and their application to research in speech and language therapy: a response to T. R. Pring's 'Ask a silly question: two decades of troublesome trials' (2004). AB - BACKGROUND: The advent of evidence-based healthcare has seen a rise in the use of systematic reviews to bring together the findings from research studies. The use of systematic reviews in speech and language therapy (SLT) was criticized in this journal in 2004 by T. R. Pring. It was claimed that their findings are misleading due to the potential inclusion of biased data, and uninformative due to a lack of detail in the reporting of interventions. It is argued that outcome research should be carried out in a series of phases in which small-scale research precedes large-scale research. This, it is argued, is most likely to demonstrate statistically significant effects and also help to ensure that therapies become sufficiently defined so that clinicians can apply them in practice. AIMS: This paper argues that the above criticism of systematic reviews is based on a narrow conception of their capabilities: on the popular misapprehension that all systematic reviews answer effectiveness questions using only experimental studies and contain meta-analyses. Different methods for systematic reviews are described and their application within clinical outcome research is discussed with reference to a phased structure for empirical enquiry. MAIN CONTRIBUTION: Systematic reviews seek to identify and synthesize information within a given topic area. They are used to answer a wide range of research questions and the studies they include are not limited exclusively to experimental designs. Methods of synthesis can include both statistical approaches, such as meta-analysis, and 'qualitative' approaches, such as meta-ethnography and thematic analysis. Knowledge of the current state of research is essential for a sequentially phased approach within outcome research to operate. Since systematic reviews are summaries of research activity, they can provide this knowledge and should therefore be considered a valuable tool within outcome research. CONCLUSIONS: A systematic review using 'qualitative' and/or statistical methods for combining studies can be carried out within or across any of the phases within outcome research. Far from being uninformative, this can help bring together what is, and what is not, known and indicate the kinds of therapies that may be beneficial in the clinical setting and therapies which would benefit from further research and development. PMID- 16272006 TI - Systematic reviews require a systematic approach to therapy research: a reply to Garrett and Thomas (2005). PMID- 16272036 TI - Obstetric risk factors and outcome of pregnancies complicated with early postpartum hemorrhage: a population-based study. AB - OBJECTIVE: The study was aimed to identify obstetric risk factors for early postpartum hemorrhage (PPH) in singleton gestations and to evaluate pregnancy outcome. STUDY DESIGN: A comparison between consecutive singleton deliveries with and without early PPH was performed. Deliveries occurred during the years 1988 2002 in a tertiary medical center. A multivariate logistic regression model was constructed in order to define independent risk factors for PPH. RESULTS: Postpartum hemorrhage complicated 0.4% (n = 666) of all deliveries enrolled in the study (n = 154 311). Significant risk factors for PPH, identified using a multivariable analysis, were: retained placenta (OR 3.5, 95%CI 2.1-5.8), failure to progress during the second stage of labor (OR 3.4, 95%CI 2.4-4.7), placenta accreta (OR 3.3, 95%CI 1.7-6.4), lacerations (OR 2.4, 95%CI 2.0-2.8), instrumental delivery (OR 2.3, 95%CI 1.6-3.4), large for gestational age (LGA) newborn (OR 1.9, 95%CI 1.6-2.4), hypertensive disorders (OR 1.7, 95%CI 1.2-2.1), induction of labor (OR 1.4, 95%CI 1.1-1.7) and augmentation of labor with oxytocin (OR 1.4, 95%CI 1.2-1.7). Women were assigned into three different groups according to the assessed severity of PPH, assuming that the severe cases were handled by revision of the birth canal under anesthesia, and the most severe cases required in addition treatment with blood products. A significant linear association was found between the severity of bleeding and the following factors: vacuum extraction, oxytocin augmentation, hypertensive disorders as well as perinatal mortality, uterine rupture, peripartum hysterectomy and uterine or internal iliac artery ligation (p < 0.001 for all variables). CONCLUSION: Hypertensive disorder, failure to progress during the second stage of labor, oxytocin augmentation, vacuum extraction and LGA were found to be major risk factors for severe PPH. Special attention should be given after birth to hypertensive patients, and to patients who underwent induction of labor or instrumental delivery, as well as to those delivering LGA newborns. PMID- 16272037 TI - The effects of standing, lifting and noise exposure on preterm birth, growth restriction, and perinatal death in healthy low-risk working military women. AB - OBJECTIVE: The effects of standing, lifting and noise in low-risk, healthy pregnant women are uncertain. In the past, the heterogeneity of the populations studied, the limitations of the designs of the retrospective and case control studies, and a failure of some of the larger investigations to evaluate all the potential confounding variables has hampered many studies. The purpose of this investigation was to evaluate, throughout pregnancy, the effects of standing, repetitive lifting, and noise in the workplace compared with no standing, lifting or noise exposure, on maternal and perinatal outcomes in a large prospective study of a low-risk healthy population of working women cared for by a single group of health providers. METHODS: This prospective observational study used an extensive questionnaire to collect antepartum, intrapartum, and postpartum information. Information was collected on the initial visit, each subsequent visit, and immediately after delivery. The participating women were divided into groups based on the amount of time spent standing, the amount and extent of repetitive lifting, and noise exposure in the workplace. RESULTS: Eight hundred and fourteen low-risk active duty women participated in this investigation over a 4-year period. Multivariate analysis with non-exposure compared with exposure reinforced the effect of standing on preterm labor (OR 1.80, 95% CI 1.05, 3.16) and preterm birth (OR 1.69, 95% CI 1.03, 2.80) and showed a trend toward an effect of noise exposure on preterm labor (OR 1.76, 95% CI 0.78, 3.39) after controlling for other exposures. CONCLUSIONS: This investigation suggests an association of occupational standing with preterm labor and preterm birth. PMID- 16272038 TI - Maternal serum ferritin concentration in patients with preterm labor and intact membranes. AB - OBJECTIVE: To determine the relationship between maternal serum ferritin concentrations in the second trimester and the risk of preterm delivery (PTD). METHODS: A prospective observational study was conducted. Fifty consecutive women with singleton pregnancies, who were admitted to the Maternal Fetal Medicine Unit due to preterm labor in the second trimester, were included. Maternal serum samples for determination of ferritin concentrations were obtained. Multiple logistic regression analysis was performed to control for confounders. RESULTS: Out of fifty patients enrolled in the study, 38% (19/50) delivered prematurely. Eight women (16%) had maternal serum ferritin concentrations above 30 ng/ml in the second trimester. Among them, 75% (n = 6) subsequently presented with preterm delivery (odds ratio (OR) = 6.7 with 95% confidence interval (CI) 1.1-56.2, p = 0.04). Only two patients with increased maternal ferritin concentrations delivered at term. However, 13 patients with second trimester ferritin concentrations below 30 ng/ml had preterm delivery. No significant differences in mean maternal ferritin concentrations were found between patients who delivered preterm and those that delivered at term, 31.9 +/- 50.6 vs. 13.6 +/- 15.2, respectively (p = 0.064). Using a multivariable analysis, controlling for anemia, leucocytosis and maternal age, increased serum ferritin concentrations were found to be an independent risk factor for PTD (OR = 8.6; 95% CI 1.4-52.5; p < 0.019). No significant correlation was found between serum ferritin concentrations and gestational age at birth (Pearson correlation coefficient r = -0.093; p = 0.522). CONCLUSIONS: Maternal ferritin concentrations above 30 ng/ml in the second trimester can serve as a marker for preterm delivery. However, since no correlation was found between serum ferritin concentrations and gestational age at birth, the routine use of serum ferritin as a marker for preterm delivery warrants further investigation. PMID- 16272039 TI - Risk of preeclampsia in relation to maternal history of migraine headaches. AB - OBJECTIVE: We examined the relationship between migraines and preeclampsia risk. STUDY DESIGN: Cases were 244 women with preeclampsia and controls were 470 normotensive women. Women were asked if a physician had ever told them that they had migraines. Odds ratios (OR) and 95% confidence intervals (CI) were calculated. RESULTS: A history of migraines was associated with a 1.8-fold increased risk of preeclampsia (95% CI 1.1-2.7). Women who were 30+ years old when diagnosed with migraines had the highest risk (OR 2.8, 95% CI 0.8-9.0). The migraine-preeclampsia association appeared to be modified by pre-pregnancy overweight status (p = 0.06). Overweight migrainous women, compared with lean nonmigrainous women, had a 12-fold increased preeclampsia risk (95% CI 5.9-25.7). CONCLUSION: Our findings are consistent with reports from six of eight previous studies on the topic. Nevertheless, prospective cohort studies are needed to further evaluate the extent to which migraines and/or its treatments are associated with preeclampsia risk. PMID- 16272041 TI - External validation of the biodegradability prediction model CATABOL using data sets of existing and new chemicals under the Japanese Chemical Substances Control Law. AB - External validation of the biodegradability prediction model CATABOL was conducted using test data of 338 existing chemicals and 1123 new chemicals under the Japanese Chemical Substances Control Law. CATABOL predicts that 1089 chemicals will have a BOD < 60% while 925 (85%) actually have an observed BOD<60%. The percentage of chemicals with an observed BOD value <60% tends to increase as the predicted BOD values decrease. In contrast, 340 chemicals were predicted to have a BOD > or = 60% and 234 (69%) actually had an observed BOD > or = 60%. The prediction of poor biodegradability was more accurate than the predictions of high biodegradability. The features of chemical structures affecting CATABOL predictability were also investigated. PMID- 16272042 TI - A new strategy for using supervised artificial neural networks in QSAR. AB - A new type of environmental QSAR model is presented for the common situation in which the biological activity of molecules mainly depends on their 1 octanol/water partition coefficient (log P). In a first step, a classical regression equation with log P is derived. The residuals obtained with this simple linear equation are then modeled from a supervised artificial neural network including different molecular descriptors as input neurons. Finally, results produced by the linear and nonlinear models are both considered for calculating the activity values, which are compared with the initial actual activity values. A heterogeneous database of 569 organic compounds with 96-h LC50s measured to the fathead minnow (Pimephales promelas), randomly divided into a training set of 484 chemicals and a testing set of 85 chemicals, was used as illustrative example to show the potentialities of this new modeling strategy Finally, practical suggestions are given for designing this type of hybrid QSAR model. PMID- 16272043 TI - Physico-chemical properties of PCDD/PCDFs and phthalate esters. AB - QSPR models for water solubility (S), n-octanol/water partition coefficient (K(OW)), and Henry's law constant (H) for polychlorinated dibenzo-p-dioxins (PCDDs) and dibenzo-p-furans (PCDFs) and phthalates have been established based on two different sets of parameters. Those parameters were topology based characteristic root index (CRI) and three semi-empirical molecular descriptors, namely--energies of the highest occupied and the lowest unoccupied molecular orbital (E(HOMO) and E(LUMO)), and dipole moment (mu). The best fit equation found by "forward multiple linear regression" showed that the topology based CRI was the most important parameter for the modelling of solubility and n octanol/water partition coefficient. For n-octanol/water partition coefficient a two-parameter equation including the CRI and E(HOMO) with a correlation coefficient of r = 0.992 was obtained whereas a three-parameter equation for solubility and Henry's law constant including the CRI, E(LUMO) and mu with a correlation coefficient of r = 0.986 and r = 0.933 was obtained, respectively. E(HOMO) and mu didn't appear in the same model because of the collinearity. The results of modified jackknife tests indicated that the three models were statistically robust. Mean deviation of calculated values from experimental data amounted to 0.27, 0.17, and 0.28 log units for the three properties mentioned. The developed models have been used to predict the S, K(OW) and H of compounds not included in the training sets. PMID- 16272044 TI - Henry's law constant of hydrocarbons in air-water system: the cavity ovality effect on the non-electrostatic contribution term of solvation free energy. AB - In this study, a quantitative structure-property relationship (QSPR) model for the prediction of Henry's law constants of aliphatic hydrocarbons in air-water system has been developed, based on a data-set of 189 compounds. The well-known linear thermodynamic relation between the logarithm of Henry's law constant and solvation free energy has been used for developing the model. It is emphasised that the solvent-accessible surface area (SASA) descriptor is not adequate for predicting the solvation free energy of a wide range of aliphatic hydrocarbons; there are many compounds that have the same solvent-accessible surface area with different solvation free energy. Therefore, we have introduced cavity ovality as a good descriptor of molecular cavity shape factor. The root mean square error (RMSE) of the QSPR regression model based on SASA improves from 0.40 to 0.22 by introducing the cavity ovality descriptor. The QSPR linear ovality model has good statistical parameters (r(2) = 0.90). To emphasise the significant effect of the new descriptor, a non-linear neural network model with only two nodes in the hidden layer was developed, and also yielded a RMSE of 0.22. PMID- 16272045 TI - Comparison of three fish bioaccumulation models for ecological and human risk assessment and validation with field data. AB - This article compares two bioconcentration Quantitative Structure Activity Relationships (QSARs) for fish applied in human risk assessments with the mechanistic bioaccumulation model OMEGA and field data. It was found that all models are virtually similar up to a Kow of 10(6). For substances with a Kow higher than 10(6), the fish bioconcentration curve in the risk assessment model EUSES decreases parabolically. In contrast, OMEGA bioaccumulation outcomes approximately show a linear increase, based on mechanistic bioconcentration and biomagnification properties of chemicals. The OMEGA-outcomes are close to the fish bioconcentration outcomes of the risk assessment model CalTOX. For very hydrophobic substances, field accumulation data in freshwater and marine fish species are closer to OMEGA- and CalTOX-outcomes compared to EUSES. The results also show that it is important to include biomagnification in fish and lipid content of fish in human exposure models. PMID- 16272046 TI - Prediction of CL-20 chemical degradation pathways, theoretical and experimental evidence for dependence on competing modes of reaction. AB - Highest occupied and lowest unoccupied molecular orbital energies, formation energies, bond lengths and FTIR spectra all suggest competing CL-20 degradation mechanisms. This second of two studies investigates recalcitrant, toxic, aromatic CL-20 intermediates that absorb from 370 to 430 nm. Our earlier study (Struct. Chem., 15, 2004) revealed that these intermediates were formed at high OH(-) concentrations via the chemically preferred pathway of breaking the C-C bond between the two cyclopentanes, thereby eliminating nitro groups, forming conjugated pi bonds, and resulting in a pyrazine three-ring aromatic intermediate. In attempting to find and make dominant a more benign CL-20 transformation pathway, this current research validates hydroxylation results from both studies and examines CL-20 transformations via photo-induced free radical reactions. This article discusses CL-20 competing modes of degradation revealed through: computational calculation; UV/VIS and SF spectroscopy following alkaline hydrolysis; and photochemical irradiation to degrade CL-20 and its byproducts at their respective wavelengths of maximum absorption. PMID- 16272047 TI - New challenges for VISION 2020. PMID- 16272048 TI - Comparison of endothelial cell loss and surgically induced astigmatism following conventional extracapsular cataract surgery, manual small-incision surgery and phacoemulsification. AB - PURPOSE: To compare the surgically induced astigmatism (SIA) and endothelial cell loss following conventional extracapsular cataract surgery (ECCE), manual small incision cataract surgery (Blumenthal technique)(SICS) and phacoemulsification (PE) with non-foldable intraocular lens implantation. METHODS: 186 cataractous eyes with nuclear sclerosis grade 3 or less were randomized to undergo ECCE, SICS or PE with intraocular lens (non-foldable) implantation after a detailed pre operative assessment. Keratometry and specular microscopy were performed pre operatively and 6 weeks postoperatively. Surgically induced astigmatism was calculated using the rectangular coordinate method (Holladay et al.). RESULTS: Mean endothelial cell loss was similar for all three groups (p = 0.855); ECCE induced a loss of 4.72% (SD: 13.07); SICS 4.21% (SD: 10.29) and PE 5.41% (SD: 10.99). Mean SIA was 1.77D (1.61D) for the ECCE group, 1.17D (0.95D) for the SICS group and 0.77D (0.65D) for the PE group (p = 0.001). The magnitude of the difference between the SICS and the PE group was 0.4D. CONCLUSION: PE induced less astigmatism than SICS and ECCE in this study but the magnitude of the difference between SICS and PE was small. There was no significant difference in endothelial cell loss between the three groups. PMID- 16272050 TI - Noncompliance with medical treatment among glaucoma patients in Oman--a cross sectional descriptive study. AB - PURPOSE: Glaucoma is a priority eye disease in Oman. In order to revise the existing policies for eye care, a study was conducted to review the noncompliance rates among glaucoma patients. METHODS: Using the random method, we selected 105 glaucoma patients from among those visiting glaucoma clinics during 2003. They were interviewed by trained health professionals. A closed-ended questionnaire with 42 questions was used to note their responses. The questions dealt with their personal characteristics along with the patients' glaucoma treatment and their knowledge of and attitudes towards different components of noncompliance with medical treatment. The field staffs also observed the 'practice' of instilling placebo eyedrops. The responses were analyzed to grade the compliance, knowledge, attitude and practice of each patient. RESULTS: Excellent compliance was found in 26 patients. Discontinuation of medication in the past was reported by 87 patients. Missing instillation of eyedrops and irregular visits to the ophthalmologist were noted in 10 and 14 patients, respectively. Adequate knowledge about glaucoma and its complications was negatively associated with noncompliance [RR 2.05 (95% CI 1.07-3.93)]. CONCLUSIONS: The high rates of noncompliance among glaucoma patients in Oman demand a systematic programmed approach. Longitudinal studies are needed to confirm the causal association between noncompliance and Knowledge, Attitude and Practice. Health education, communication while dispensing medication, reducing the frequency of medication and using devices for the easy administration of eyedrops could further improve compliance. PMID- 16272049 TI - Primary open-angle glaucoma presentation at a tertiary unit in Africa: intraocular pressure levels and visual status. AB - AIM: To investigate the presentation of POAG at a tertiary referral hospital in East Africa, including intraocular pressures, visual status and management decisions. METHODS: Retrospective review of first-time presenters with POAG over a 6-month period. RESULTS: Of the 298 patients identified, mean age 57 years (n = 296, range = 14-88, SD = 14), 213 (72%) male, 122 (41%) had normal vision (using WHO better eye acuity criteria: visual impairment < 6/18, blind < 3/60) at presentation, 87 (30%) had visual impairment and 86 (29%) were blind. The mean presenting IOP was 32 mmHg (SD = 11) and 70% of the patients had a cup:disc ratio of 0.8 or worse in the better eye. Longer disease duration (OR = 1.20, 95% CI 1.04-1.39) and higher mean IOP (OR = 1.06, 95% CI 1.02-1.10) were associated with visual impairment or blindness. Intraocular pressure showed a negative linear correlation with presenting logMAR acuity (R(2) = 0.115, SE = 1.30, p < 0.0005). The mean IOP in eyes that had undergone trabeculectomy (19 mmHg, SD = 8, n = 17) was significantly (p < 0.0005) lower than that in eyes that had not (34 mmHg, SD = 12, n = 274). The referral rate for trabeculectomy was 158/275 (57%). Male sex (OR = 2.17, 95% CI 1.0-4.72), higher mean IOP (OR = 1.09, 95% CI 1.05-1.14) and not being blind (OR = 26.47, 95% CI 9.90-70.78) were associated with surgical referral. Of the 158 patients, 76 (48%) accepted surgery. CONCLUSIONS: A high proportion of patients presenting to our unit with POAG are visually impaired or blind and the higher their presenting IOP the poorer their visual acuity. Previous trabeculectomy was associated with lower IOPs and protection from visual impairment and blindness. Further training of clinic staff towards early surgical referral and investigation of gender barriers is required. PMID- 16272051 TI - The problem of trachoma within an onchocerciasis-endemic zone in Kaduna: need for a multilateral approach? AB - INTRODUCTION: Within the mesoendemic onchocerciasis belt of Kaduna State, Northern Nigeria, there were other causes of ocular morbidity as well. We investigated the contribution of trachoma. METHODS: A total of 6831 individuals were examined for eye disease, including signs of trachoma. RESULTS: The overall prevalence of trachoma was 4.5% (577 eyes of 310 individuals), making it the second most common cause (1 in 5) of ocular morbidity after cataracts. Of the 577 eyes, 405 (70.2%) had cicatrizing disease while the rest had active disease. Overall, the prevalence of TF was 1.1% (1.3% in those < 10 years of age), TI 0.2%, TS 3%, CO 0.5% and TT 1.1% (4.3% and 3.5% in subjects aged > 55 and > 40 years, respectively). The prevalence of trachoma increased with age. The overall prevalence of blindness in the whole population was 2.7%. Trachoma accounted for 11% of this. On the other hand, onchocerciasis accounted for 3.1% of the overall ocular morbidity (less than trachoma) and 39% of the blindness. CONCLUSIONS: There appears to be a clear need to retrain middle cadre ivermectin workers in trachoma rapid assessment and SAFE strategy intervention methods in order to further impact blindness prevention in these 'onchocerciasis-endemic' zones. PMID- 16272053 TI - Risk factors for postoperative hemorrhage after vitrectomy for diabetic retinopathy. AB - OBJECTIVE: To identify the potential risk factors for postvitrectomy diabetic vitreous hemorrhage (PDVH). METHODS: A matched case-control nested into a retrospective follow-up study was done to review the surgical results in 68 consecutive eyes undergoing primary pars plana vitrectomy for vitreous hemorrhage. The eyes were divided into two groups based on the presence of PDVH (19 cases and 49 controls) and were matched on surgeon and the date of surgery. Twenty-three factors related to the preoperative examination and eight factors related to the operative procedure were analyzed. Statistical analysis was based on conditional logistic regression models with PDVH as the dependent variable. The mean follow-up interval was six months. RESULTS: The factors associated with the incidence of PDVH were iris neovascularization (OR = 9.8, P = 0.03), lower extremity amputations (OR = 8.3, P = 0.02) and the use of antihypertensive agents within three months before vitrectomy (OR = 0.2, P = 0.04). Phakic and aphakic eyes of diabetic patients with lower extremity amputations would have a 70% probability of developing PDVH. This probability would have dropped to 30-40% had they been taking antihypertensive treatment. CONCLUSIONS: Iris neovascularization and lower extremity amputations increase the risk of PDVH. Antihypertensive treatment before vitrectomy decreases this risk. PMID- 16272052 TI - The Pakistan national blindness and visual impairment survey--research design, eye examination methodology and results of the pilot study. AB - PURPOSE: To establish age- and sex-specific prevalence rates and causes of blindness and low vision in children aged 10 to 15 years and adults aged 30 years and older in Pakistan. METHODS: Multi-stage, stratified (rural/urban), cluster random sampling, with probability proportional-to-size procedures, was utilised to select a cross-sectional, nationally representative sample of adults (16,600 subjects) and children (6,000 subjects). Each subject underwent: interview, visual acuity (logMAR), autorefraction and optic disc examination. Those that saw < 6/12 in either eye underwent corrected visual acuity and dilated posterior segment examination. RESULTS: The results of a pilot survey are reported in this paper. In the two rural pilot sites, 159 subjects (including 47 children) were examined; 50% were male. Thirty seven adults (23.3%) but no children saw worse than 6/12 in either eye. Two subjects were blind (corrected visual acuity) in the better eye, and 11 were visually impaired. Refractive error was the main cause (in 22 eyes (39% of the total of 56 eyes)) of < 6/12 visual acuity, followed by cataract (12 eyes), uncorrected aphakia (6 eyes) and age-related macular disease (3 eyes). CONCLUSIONS: The pilot survey demonstrated that the proposed examination process for the main survey is feasible. Particular strengths of this survey include the use of logMAR visual acuity testing and autorefraction of all subjects, a dilated posterior segment examination, and the use of a 'less than 6/12' threshold for further examination. This lower threshold addresses the burden of refractive error, which, with cataract, are two of the diseases specifically targeted by Vision 2020. PMID- 16272055 TI - Juvenile X-linked retinoschisis with normal scotopic b-wave in the electroretinogram at an early stage of the disease. AB - PURPOSE: To report four cases of genetically verified juvenile X-linked retinoschisis (XLRS) with normal scotopic b-waves in full-field ERG, including one patient with a novel mutation (W50X) in the RS1 gene. METHODS: Four XLRS patients from different families were examined with regard to visual acuity, kinetic perimetry, fundus photography, full-field ERG, and OCT. Two of these patients were also examined with multifocal-ERG (mfERG). Mutations in the RS1 gene were identified by sequence analysis. RESULTS: The full-field ERG presented normal b-wave amplitudes on scotopic white-light stimulation. OCT and mfERG presented macular schisis and macular dysfunction. Genetic analysis revealed a deletion of exon 1 and the promotor region in one patient and mutations giving rise to the amino acid substitutions R209C and W96R in two others. The fourth patient carried a novel mutation in exon 3 of the RS1 gene (nt 149 G-->A), causing the introduction of a stop codon after amino acid 49 in the RS protein. CONCLUSION: Four young males with XLRS did not present with reduction in the scotopic b-wave amplitude on full-field ERG, which is otherwise often considered to be characteristic of the disease. Full-field ERG and molecular genetic analysis of the RS1 gene still remain the most important diagnostic tools for this retinal disorder, although the OCT can be a valuable complement in order to make the diagnosis at an early stage. PMID- 16272054 TI - Cost-effectiveness analysis of PMMA, silicone, or acrylic intra-ocular lenses in cataract surgery in four European countries. AB - PURPOSE: To compare the cost-effectiveness of different intra-ocular lens (IOL) materials (Hydrophobic acrylic, Polymethylmethacrylate (PMMA), Hydrophilic acrylic and Silicone) implanted after cataract surgery with reference to Nd:YAG laser capsulotomy and Nd:YAG-related complications in four European countries (France, Italy, Germany and Spain). SETTING: A retrospective review of 1,525 patients (eyes), aged 50 to 80 years, operated with phacoemulsification for cataract in 1996 or 1997 in 16 surgical centres (4 per country). METHODS: The study was conducted using a cost-effectiveness approach. Medical charts were reviewed to collect retrospective information during the 3-year period following cataract surgery in order to identify patients who underwent Nd:YAG laser capsulotomy post-operatively. Clinical data were combined with unit costs assessed by experts for Nd:YAG laser capsulotomy and their complications. A cost effectiveness ratio (cost per patient without Nd:YAG laser capsulotomy intervention) was estimated in relation to each IOL material used in each of the four European countries. RESULTS: Hydrophobic acrylic, specifically Acrysof, was the most cost-effective IOL material in all the countries except Germany where it was second. PMMA had the best ratio in Germany, was second in Spain and only third in Italy and France. Silicone was second in France and ranked third in the other countries, while hydrophilic acrylic had the worst ratio overall in all countries. CONCLUSIONS: Cost-effectiveness ratios of hydrophobic acrylic (Acrysof) were better than those of other types of IOL materials used in most of the countries. Sensitivity analyses were performed to vary the base case analysis to demonstrate the economic importance of the assumptions. In all cases, hydrophobic acrylic IOL material was shown to be a highly cost-effective option. PMID- 16272056 TI - Clinical phenotype in a Swedish family with a mutation in the IMPDH1 gene. AB - PURPOSE: Mutations in the inosine monophosphate dehydrogenase 1 gene (IMPDH1) have recently been discovered to cause a form of autosomal dominant retinitis pigmentosa (adRP). Such mutations are estimated to account for approximately 2-5% of the adRP cases among Americans of European origin and Europeans. Aiming towards an understanding of the molecular background of retinitis pigmentosa, this paper describes the phenotype of a Swedish family with a mutation in IMPDH1. METHODS: Venous blood samples were obtained from 12 family members and screened for mutations in IMPDH1. Six individuals with the mutation were examined clinically and with full-field electroretinography (ERG), dark adaptometry, multifocal electroretinography (mfERG), and optical coherence tomography (OCT). Also reviewed were the clinical findings and ERGs obtained 14 years earlier. RESULTS: The proband and eight other relatives from three generations were found to harbor the Asp226Asn mutation in IMPDH1. These individuals, from three generations, showed clinical and electrophysiological signs of retinitis pigmentosa. The cone responses to the full-field, 30-Hz flicker ERG demonstrated an unusual pattern, with implicit times within normal limits or only slightly prolonged. Rod ERG responses, however, were undetectable. OCT showed intraretinal fluid and swelling, changes that were more pronounced in younger individuals. mfERG showed residual preserved central function. The older the individual, the smaller the area of preserved central function. CONCLUSION: In this family with a mutation in IMPDH1, we found a specific phenotype with rod function affected more than cone function, foveal edema, and central retinal function preserved for a long period of time. Foveal edema could be a pathogenic feature in this form of retinal degeneration. PMID- 16272057 TI - Posterior polar cataract: genetic analysis of a large family. AB - Congenital cataracts are clinically and genetically heterogeneous. Loci for autosomal dominant posterior polar cataracts have been mapped to chromosomes 1p36, 11q22-q22.3, 16q22, and 20p12-q12. We investigated a large four-generation family with 20 individuals affected with congenital posterior polar cataracts. After exclusion of known loci for posterior polar cataracts, a genome-wide screen was conducted. In this family, we mapped dominant congenital posterior polar cataracts to chromosome 10q24. On haplotype analysis, we identified an 11-cM interval between loci D10S1680 and D10S467, which included the PITX3 gene. On sequencing the coding region of PITX3, we found a 17-base-pair duplication in exon 4. Although the same genotype was described in a family with ASMD and cataracts, the common phenotype of this mutation is probably posterior polar cataract; a modifier gene is presumed to cause anterior segment abnormalities in the previously described patients. The same mutation was recently identified in four families with congenital cataracts. This study provides further evidence of genetic heterogeneity of autosomal dominant posterior polar cataract. PMID- 16272058 TI - A new case of oculoectodermal syndrome. AB - An 11-month-old infant girl presented with right-sided features of aplasia cutis congenita of the scalp, unilateral epibulbar dermoids, eccentric pupil, coloboma of the right upper eyelid, and depigmentation of the fundus surrounding the right optic nerve. These findings were similar to the oculoectodermal syndrome reported by other clinicians and researchers. PMID- 16272059 TI - Rieger's anomaly and other ocular abnormalities in association with osteogenesis imperfecta and a COL1A1 mutation. AB - A patient with osteogenesis imperfecta (OI) and some features of Ehlers-Danlos syndrome had Rieger's anomaly and other associated ocular abnormalities. He carried a COL1A1 mutation (c.3313delA) that has only rarely been seen in OI. The association of ocular anterior chamber abnormalities with OI has not been reported previously, while OI with Ehlers-Danlos syndrome features has only been described in some kindreds. The patient had serious complications as a result of his ocular anomalies. We speculate that the course of his disease and, perhaps, its co-existence with OI could be exacerbated by his collagen type-I defect, although no causality can be established by this report of a single case. PMID- 16272061 TI - Ophthalmic aspects of GAPO syndrome: case report and review. AB - This paper reports on a 36-year-old woman with GAPO syndrome, a rare autosomal recessive condition characterized by growth retardation (G), alopecia (A), pseudoanodontia (P), and optic atrophy (O). Her parents are consanguineous and one of her sisters is also affected. Since the first description by Anderson and Pindborg in 1947, 27 individuals have been reported with this diagnosis. They were from at least 19 different families (four of them from Brazil, including the present one), suggesting a founder effect. The phenotype of this condition, initially considered as the result of an ectodermal dysplasia, could be attributed to the accumulation of extracellular connective tissue matrix and its progressive character must be pointed out. The clinical findings, especially ophthalmological features that include bilateral glaucoma, are reviewed and discussed. PMID- 16272060 TI - Mild optic nerve hypoplasia with retinal venous tortuosity in aarskog (facial digital-genital) syndrome. AB - Aarskog syndrome (faciogenital dysplasia) is an X-linked recessive genetic growth disorder characterized by short stature, dysmorphic facies, shawl scrotum, and digital anomalies. The condition was first described in 1970 and the gene responsible is FGD1 (MIM#305400). There are several reported ophthalmic findings associated with Aarskog syndrome which are discussed. We describe a case of Aarskog syndrome with venous tortuosity, optic nerve hypoplasia, and a type-2 antithrombin deficiency. PMID- 16272064 TI - The Impact Factor--a dubious measure of scientific quality. PMID- 16272063 TI - Association Down syndrome-retinoblastoma: a new observation. PMID- 16272065 TI - News from the Nordic Colleges of General Practitioners. PMID- 16272066 TI - Job satisfaction among Norwegian general practitioners. AB - OBJECTIVE: To explore the level of job satisfaction among general practitioners (GPs) and to compare it with that of hospital doctors. DESIGN: Postal questionnaire among Norwegian doctors in 2002 and similar data from 1994 and 2000 for most of the respondents. MATERIAL: A total of 295 GPs out of 1174 doctors completed the questionnaire (73% response rate). Main outcome measures. Self reported levels of job satisfaction according to the Job Satisfaction Scale (JSS). RESULTS: Norwegian GPs reported a high level of job satisfaction with a mean score on the JSS of 52.6 (10 is minimum and 70 maximum). The reported level of satisfaction was highest for their opportunities to use their abilities, cooperation with colleagues and fellow workers, variation in work, and freedom to choose own method of working. The GPs' level of job satisfaction remained stable in 1994, 2000, and 2004 in spite of major health reforms. GPs report a higher level of job satisfaction than hospital doctors. CONCLUSIONS: In spite of international discussions on unhappy doctors and doctors' discontent, Norwegian GPs do report a high and stable level of job satisfaction. PMID- 16272067 TI - Characteristics of middle-aged and elderly women with urinary incontinence. AB - OBJECTIVE: To describe the prevalence of urinary incontinence and to find characteristics useful in general practice for identifying middle-aged and elderly women with the problem. DESIGN: Cross-sectional interview study. SETTING: Population-based samples of Danes. SUBJECTS: A total of 5795 women older than 45 years (46+ years). MAIN OUTCOME MEASURES: Prevalence of incontinence and clinical characteristics assessed by standardized interview questions. RESULTS: The overall prevalence of urinary incontinence was 20% among women less than 60 years of age and 44% among those older than 80 years. Increasing age was highly associated with both forms of incontinence (stress and urge). High body mass index (BMI), chronic lung disease, and stroke were also associated with both forms, while number of children was associated with stress incontinence only. Predictive models show that 56% of women characterized by high age (older than 80 years) and overweight (BMI higher than 30) will suffer from urinary incontinence. The low-risk group defined by these two parameters (aged 46-60 years and not overweight) still had a 19% prevalence in the last month. CONCLUSION: The prevalence of urinary incontinence increased with age. Even in the low-risk groups the problem was very common in old age. Questions about incontinence should, therefore, be asked in relevant consultations with all elderly female patients. PMID- 16272068 TI - Health problems associated with lower urinary tract symptoms in older women. A population-based survey. AB - OBJECTIVE: To identify health problems associated with lower urinary tract symptoms (LUTS) in an older female population. DESIGN: Population-based cross sectional survey. SETTING: Third wave for the Tampere Longitudinal Study on Ageing (TamELSA) in 1999-2000. SUBJECTS: A total of 203 women aged 70 years and over. MAIN OUTCOME MEASURES: LUTS categorized as urge symptoms only, voiding symptoms only, and urge and voiding symptoms combined. The health indicators were polypharmacy (> 3 medications), use of sleeping medication, constipation, treated urinary tract infections (UTIs), and neurological, cardiovascular, and musculoskeletal diseases. RESULTS: Of the respondents 16% reported urge symptoms, 22% voiding symptoms, and 26% combined symptoms. In the multinomial logistic regression models, constipation and musculoskeletal diseases were significantly associated with voiding symptoms (OR 4.33; 95% CI 1.62-11.57 and OR 5.25; 95% CI 1.94-14.18, respectively) and with combined symptoms (OR 4.33; 95% CI 1.67-11.21 and OR 2.84; 95% CI 1.17-6.89, respectively). UTIs (OR 2.78; 95% CI 1.09-7.08) were associated with combined symptoms. CONCLUSIONS: Voiding symptoms comprise a significant part of lower urinary tract symptomalogy in older women. Although constipation, musculoskeletal diseases, and UTIs are the main health problems associated with LUTS, other concomitant diseases and medications also need to be assessed in older female patients with LUTS. PMID- 16272069 TI - Balance performance and self-perceived handicap among dizzy patients in primary health care. AB - OBJECTIVE: To study the diagnostic panorama at a primary health care centre where the physiotherapist is specialized in dizziness. To study balance measures of dizzy patients as well as measures of self-perceived handicap and to analyse whether these measures correlate. DESIGN: Retrospective study of computerized medical records. SETTING: A primary health care centre in Malmo, Sweden. SUBJECTS: A total of 119 patients with dizziness, 73 women and 46 men, aged from 22 to 90 years. MAIN OUTCOME MEASURES: Diagnoses according to specified criteria. Four balance measures: tandem standing, standing on one leg, walking in a figure of eight, and walking heel to toe on a line. The Dizziness Handicap Inventory (DHI). RESULTS: Six different groups of diagnoses were found: multisensory dizziness, peripheral vestibular disorder, dizziness as a symptom caused by whiplash-associated disorder, unspecific dizziness, phobic postural vertigo, and dizziness of cervical origin. The group with multisensory dizziness performed poorer on the balance measures than the other groups. The group with phobic postural vertigo had the highest total scores on DHI, while the vestibular group had the lowest total score. Subjects over 65 years old had more disturbances in balance, but a lower level of self-perceived handicap, than subjects aged 65 or younger. DHI did not correlate with any of the balance measures. CONCLUSIONS: Self-perceived handicap, measured with DHI, and disturbed balance measured with clinical methods, do not necessarily correlate. Elderly patients with dizziness seem to have more disturbances in balance than younger patients but a lower level of self-perceived handicap. PMID- 16272070 TI - Identification and diagnostic evaluation of possible dementia in general practice. A prospective study. AB - OBJECTIVE: To investigate the rate of diagnostic evaluation of dementia for patients in whom a suspicion of dementia was raised, and to investigate reasons why a diagnostic evaluation was not always being performed. DESIGN: A prospective study among elderly patients aged 65+, and a follow-up study. SETTING: In all, 17 general practices in Copenhagen with 40 865 patients on their lists of whom 2934 were aged 65+. SUBJECTS: A total of 793 patients consulting their GP regardless of reason of encounter, in October and November 2002. MAIN OUTCOME MEASURES: MMSE score < or = 23, GP clinical impression of dementia, laboratory-screening tests prescribed by the GPs and referral status after 6 months, and follow-up questionnaire. RESULTS: Of 793 patients a total of 138 patients were identified with possible dementia. Among the identified patients 26 (20%) were referred for further evaluation within 6 months, and 4 (3%) were treated for depression or referred for another condition. A total of 6 patients were lost to follow-up. In the remaining 102 undiagnosed patients the main reasons for not performing a diagnostic evaluation of dementia were patient/relative hesitation (34%), the GP thought that it would not have any consequences for the patient, or the GP estimated that the patient was too fragile (21%). CONCLUSION: In 17% of elderly patients in general practice a suspicion of dementia could be raised based on the clinical impression of the GP or MMSE score. However, only 23% of this group were evaluated by their GP or referred to a memory clinic within a subsequent period of 6 months. PMID- 16272071 TI - Cost of heart failure in Swedish primary healthcare. AB - OBJECTIVES: To calculate the cost for patients with heart failure (HF) in a primary healthcare setting. DESIGN: Retrospective study of all available patient data during a period of one year. SETTING: Two healthcare centers in Linkoping in the southeastern region of Sweden, covering a population of 19 400 inhabitants. SUBJECTS: A total of 115 patients with a diagnosis of HF. MAIN OUTCOME MEASURES: The healthcare costs for patients with HF and the healthcare utilization concerning hospital days and visits to doctors and nurses in hospital care and primary healthcare. RESULTS: The mean annual cost for a patient with HF was SEK 37 100. There were no significant differences in cost between gender, age, New York Heart Association functional class, and cardiac function. The distribution of cost was 47% for hospital care, 22% for primary healthcare, 18% for medication, 5% for nursing home, and 6% for examinations. CONCLUSION: Hospital care accounts for the largest cost but the cost in primary healthcare is larger than previously shown. The total annual cost for patients with HF in Sweden is in the range of SEK 5.0-6.7 billion according to this calculation, which is higher than previously known. PMID- 16272072 TI - Long-term impact of elevated cardiovascular risk detected by screening. A qualitative interview study. AB - OBJECTIVE: To explore how persons with an elevated cardiovascular risk score (CRS) balanced health-related advice against the life they wanted to live or were able to live. SETTING: 2000 Danes aged 30-50 were invited to participate in a health-screening project in general practice. Screenings were conducted at baseline and after one and five years, and included among other screening procedures a calculation of CRS (see Figure 1). DESIGN: Participants with an elevated CRS were asked to participate in a qualitative semi-structured interview. They were selected by stratified purposeful sampling reflecting variations in age, sex. and perceived health. SUBJECTS: Nine men and five women aged 33-50 years. THEORETICAL FRAMES OF REFERENCE: Bandura's theory of self efficacy and the Health Belief Model's consideration of individuals' cues to act against a health threat supported analysis. RESULTS: Being informed about an elevated CRS had a considerable impact on the informants. They initiated significant lifestyle changes, though only to a limited degree when such changes would affect their quality of life adversely. In cases where other results of the multiphasic screening were normal, interpreted as such, or if there were stressful circumstances in the informant's life, the elevated CRS receded into the background. INTERPRETATION: Doctors, who inform individuals about the impact of risk factors, need to know that the consequences and health advice are not always interpreted by laypeople as supposed by the medical culture. PMID- 16272073 TI - Treatment practice in chronic epicondylitis: a survey among general practitioners and physiotherapists in Uppsala County, Sweden. AB - OBJECTIVE: To investigate treatment practice among general practitioners (GPs) and physiotherapists (PTs) in chronic epicondylitis of 3 months' duration or more. DESIGN: Postal survey. SETTING AND SUBJECTS: All 129 GPs and all 77 PTs at 35 primary health care centres in Uppsala County, Sweden, received the questionnaire. MAIN OUTCOME MEASURES: Proportion of responders using various treatments (five specified alternatives + open question). RESULTS: The questionnaire was answered by 70% of the GPs and 61% of the PTs. Ergonomic counselling, stretching, and orthotic devices were common, and used to a similar extent by GPs and PTs. Acupuncture was also common, but less so among GPs than PTs. Transcutaneous electric nerve stimulation was used by relatively few GPs and PTs. The open question revealed that dynamic exercise, particularly eccentric, was used by most PTs but only one GP. A majority of GPs prescribed sick leave and anti-inflammatory treatment with an NSAID or cortisone injections. CONCLUSION: A large number of treatment methods in chronic epicondylitis were reported, none of which is properly evidence-based and some of which are even known to be ineffective. There is a need for randomized controlled studies of potentially effective treatments in this condition. PMID- 16272074 TI - Why did I get chronic fatigue syndrome? A qualitative interview study of causal attributions in women patients. AB - OBJECTIVES: To explore causal attributions among women with chronic fatigue syndrome (CFS). DESIGN: Qualitative study where data from individual semi structured interviews were analysed according to Malterud's systematic text condensation. SETTING: Bergen, Norway. SUBJECTS: A purposeful sample of eight women aged 25-55, recruited among members of a self-help organization. MAIN OUTCOME MEASURES: Accounts of causal attribution for CFS among the informants, focusing on gender. RESULTS: The participants agreed that their way of living could have increased the vulnerability of their resistance resources. Pressure they put upon themselves, workload burdens without subsequent relaxation, emotional conflicts, or preparing for assumed problem-solving were mentioned as gendered dimensions. They presented different explanations regarding potential triggers encountering their fragile immune systems, most often a virus infection. The participants thought women might have a weaker immune system than men, or that CFS was caused by a virus that women are more likely to catch. In their experience, their symptoms were activated when people put pressure on them, such that they might be nervous as to whether they could live up to the demands of their surroundings, and in the case of emotional strain related to family and work. CONCLUSION: More studies are needed exploring hypotheses concerning the complex interplay between molecular predispositions and more or less gendered lifestyle issues in CFS. Doctors need to challenge their strong beliefs regarding medically unexplained conditions, where facts still remain unresolved. Recognizing this, the doctor may provide realistic support and advice, and contribute to the establishment of common ground for understanding and managing the condition. PMID- 16272075 TI - What is the future of primary care research? Probably fairly bright, if we may believe the historical development. AB - OBJECTIVE: To study one aspect of the development of primary care research from 1975 to 2003. DESIGN: Quantitative bibliometric study. SETTING: Pub Med database. SUBJECTS: Four Nordic countries (Denmark, Finland, Norway, and Sweden), seven countries from the rest of Europe (Belgium, France, Germany, Italy, the Netherlands, Spain, and the UK), and seven countries from the rest of the world (Australia, Canada, India, Japan, New Zealand, South Africa, and the USA). MAIN OUTCOME MEASURES: Number of primary healthcare publications per million inhabitants. Percentage of publications in primary healthcare of all publications in human medicine. RESULTS: In 2003, New Zealand, the UK, and Australia were in the lead, with barely 20 primary care publications per million inhabitants, followed by Norway, Sweden, the Netherlands, and Denmark, where the corresponding figure was around 10. A vigorous increase in publications from 1975 to 2003 was clearly seen in most of the countries. However, during the same period the proportion of publications from primary care in relation to all publications in human medicine was only moderately increased, or virtually unchanged. CONCLUSION: It is believed that primary care research has a future, and it is hoped it may even be bright. However, searching Pub Med gave but one aspect of the historical development, and in particular the comparisons between countries may be questionable. PMID- 16272076 TI - Treatment challenges in the acute hospital setting. PMID- 16272078 TI - Bipolar disorder, antidepressants and induction of hypomania or mania. A systematic review. AB - OBJECTIVE: The literature cautions against the induction of (hypo)mania owing to the use of antidepressants in bipolar disorder. Objectives of this review are to examine: (1) the evidence for this assumption; (2) underlying risk factors; and (3) the extent to which a mood stabilizer may be protective. METHOD: A systematic literature review was conducted. RESULTS: Thirteen relevant studies were included. All of them had methodological shortcomings. Overall, there is no strong evidence that use of antidepressants in bipolar disorder increases the risk of (hypo)mania. Possible, although unreplicated, risk factors are: a short allele of the promoter region of the serotonin transporter gene SCL6A4, substance abuse or dependence, multiple antidepressant trials, lower number of previous manias, less delusions during illness, depressive polarity at illness onset, and rapid cycling that has, however, been contradicted by another study. Subtype of bipolar disorder (I or II) has been considered in four studies, with conflicting results. Mood stabilizers are possibly protective. CONCLUSION: There is an urgent need for adequate studies of sufficient size. For the time being, treatment of bipolar depression may best be based on the results of the life chart of the individual patient keeping in mind the risk factors found until now. PMID- 16272077 TI - Whether ionizing radiation is a risk factor for schizophrenia spectrum disorders? AB - The neural diathesis-stressor hypothesis of schizophrenia, where neurobiological genetic predisposition to schizophrenia can be provoked by environmental stressors is considered as a model of the effects of exposure to ionizing radiation. Analysis of information from electronic databases (MEDLINE, PsycINFO, EMBASE, Current Contents, Elsevier BIOBASE) and hand-made search was carried out. There are comparable reports on increases in schizophrenia spectrum disorders following exposure to ionizing radiation as a result of atomic bombing, nuclear weapons testing, the Chernobyl accident, environmental contamination by radioactive waste, radiotherapy, and also in areas with high natural radioactive background. The results of experimental radioneurobiological studies support the hypothesis of schizophrenia as a neurodegenerative disease. Exposure to ionizing radiation causes brain damage with limbic (cortical-limbic) system dysfunction and impairment of informative processes at the molecular level that can trigger schizophrenia in predisposed individuals or cause schizophrenia-like disorders. It is supposed that ionizing radiation can be proposed as a risk factor for schizophrenia spectrum disorders. The hypothesis that ionizing radiation is a risk factor for schizophrenia spectrum disorders can be tested using data from the Chernobyl accident aftermath. Implementation of a study on schizophrenia spectrum disorders in Chernobyl accident victims is of significance for both clinical medicine and neuroscience. PMID- 16272079 TI - Do panic-agoraphobics overestimate distances? AB - Twenty-five subjects suffering from Panic Disorder with agoraphobia (PDA) and 25 normal controls (NC) were asked to estimate distances of familiar places of their town and non-distance stimuli. PDA subjects also completed the Mobility Inventory for Agoraphobia (MIA). Subjects with PDA significantly overestimated distances as compared to NC, whereas no significant difference was found on estimates of non distance stimuli. Within the PDA group, significant correlations were found between distance estimates and subsets of MIA concerning the ways of moving through the environment. Although with caution, given the small sample size and the preliminary character of this study, the results could be interpreted as consistent with a cognitive dysfunction associated with PDA, probably at the level of representational mechanisms of the extrapersonal space. PMID- 16272080 TI - Occurrence and treatment of depressive comorbidity/cosyndromality in schizophrenic psychoses: conceptual and treatment issues. AB - Depressive symptoms are a common feature of schizophrenic disorders, a fact that has become increasingly apparent over the last two decades. Apparently the introduction of standardized rating scales in cross-sectional and longitudinal investigations played an important role in the recognition of the relevance of depressive symptoms. They can be interpreted as being cosyndromal or comorbid, depending on the conceptual perspective applied. This is not simply a difference in terminology but is of great aetiopathogenetic relevance. Of particular clinical relevance is the observation that schizophrenic patients with concomitant depressive symptoms have a greater risk of suicidality or an unfavourable disease course. For this reason it is important that sufficient attention is paid to the diagnosis and treatment of depressive symptoms occurring during schizophrenic psychoses. Besides treatment with antidepressants, modern neuroleptics are of great importance in this context as they are more efficacious than classical neuroleptics in treating depressive symptoms. PMID- 16272081 TI - Long-term behavioural changes after unilateral stereotactic cingulotomy in a case of therapy-resistant alcohol dependence. AB - We report the case of a female patient who had undergone a unilateral stereotactical anterior cingulotomy (AC) 20 years previously because of refractory alcohol dependence. After AC, the drinking behaviour switched from a chronic to an episodic one with fast losses of control accompanied by abnormal oral impulse-control behaviour. Relapses were stress- but not cue-induced and followed by long-term intervals of abstinence. In addition to an intended lesion of the left dorsal anterior cingulate cortex (ACC), MR images showed additional lesions within the left caudate body and the dorsal medial thalamic nucleus. Applying a neuropsychological test battery, we found disturbed divided attention and impaired executive function. From this, we conclude that the complex lesion pattern may contribute to some of the behavioural changes seen in our patient after AC. This hypothesis is supported by the neuropsychological deficits and the fact that the neuronal circuits, impaired by the lesions, are involved in addiction-specific behaviour. This case report further emphasizes the key role of the ACC and its connections in the maintenance of dependent behaviour. PMID- 16272082 TI - Schizophrenia with catatonic features deserves further study. PMID- 16272083 TI - Pedagogy meets neuroscience. PMID- 16272084 TI - Evolution. Groups wield copyright power to delay Kansas standards. PMID- 16272085 TI - Chemistry. 'Grandfather of Nanotech' dies at 62. PMID- 16272086 TI - Biodefense. Critics question proposed countermeasures agency. PMID- 16272087 TI - Nuclear physics. Dueling experiments close in on source of proton's spin. PMID- 16272088 TI - Scientific misconduct. MIT terminates researcher over data fabrication. PMID- 16272089 TI - Science and the law. Thomas Butler loses appeal, vows to fight on. PMID- 16272091 TI - Patents. Antibody drug dispute ends in $255 million cash payout. PMID- 16272090 TI - Neuroscience. Genes that guide brain development linked to dyslexia. PMID- 16272092 TI - Italian universities. Government wins fight to modernize academic appointments. PMID- 16272093 TI - Geology. Tracking myth to geological reality. PMID- 16272094 TI - Molecular biology. P-bodies mark the spot for controlling protein production. PMID- 16272095 TI - Cancer biology. A new cancer player takes the stage. PMID- 16272097 TI - Education. Forging a cosmic connection between students and science. PMID- 16272096 TI - Synthetic biology. Synthetic biology remakes small genomes. PMID- 16272098 TI - University investment in drug discovery. PMID- 16272099 TI - A place at the pharma table for women? PMID- 16272100 TI - Costs and benefits of regulating mercury. PMID- 16272102 TI - Comment on "A brief history of seed size". PMID- 16272101 TI - Landscape corridors: possible dangers? PMID- 16272103 TI - Global voices of science. Teaching evolution in Mexico: preaching to the choir. PMID- 16272104 TI - Neuroscience. The neuron doctrine, redux. PMID- 16272106 TI - Atmospheric science. Water vapor feedback in climate models. PMID- 16272105 TI - Structural biology. A ribosomal coup: E. coli at last! PMID- 16272107 TI - Chemistry. Building a quintuple bond. PMID- 16272108 TI - Eppendorf 2005 winner. A Drosophila OBP required for pheromone signaling. PMID- 16272111 TI - Friendly faces and unusual minds. PMID- 16272112 TI - Patterning and plasticity of the cerebral cortex. AB - The cerebral cortex of the human brain is a sheet of about 10 billion neurons divided into discrete subdivisions or areas that process particular aspects of sensation, movement, and cognition. Recent evidence has begun to transform our understanding of how cortical areas form, make specific connections with other brain regions, develop unique processing networks, and adapt to changes in inputs. PMID- 16272113 TI - Map plasticity in somatosensory cortex. AB - Sensory maps in neocortex are adaptively altered to reflect recent experience and learning. In somatosensory cortex, distinct patterns of sensory use or disuse elicit multiple, functionally distinct forms of map plasticity. Diverse approaches-genetics, synaptic and in vivo physiology, optical imaging, and ultrastructural analysis-suggest a distributed model in which plasticity occurs at multiple sites in the cortical circuit with multiple cellular/synaptic mechanisms and multiple likely learning rules for plasticity. This view contrasts with the classical model in which the map plasticity reflects a single Hebbian process acting at a small set of cortical synapses. PMID- 16272114 TI - Language acquisition and brain development. AB - Language acquisition is one of the most fundamental human traits, and it is obviously the brain that undergoes the developmental changes. During the years of language acquisition, the brain not only stores linguistic information but also adapts to the grammatical regularities of language. Recent advances in functional neuroimaging have substantially contributed to systems-level analyses of brain development. In this Viewpoint, I review the current understanding of how the "final state" of language acquisition is represented in the mature brain and summarize new findings on cortical plasticity for second language acquisition, focusing particularly on the function of the grammar center. PMID- 16272115 TI - Sex differences in the brain: implications for explaining autism. AB - Empathizing is the capacity to predict and to respond to the behavior of agents (usually people) by inferring their mental states and responding to these with an appropriate emotion. Systemizing is the capacity to predict and to respond to the behavior of nonagentive deterministic systems by analyzing input-operation-output relations and inferring the rules that govern such systems. At a population level, females are stronger empathizers and males are stronger systemizers. The "extreme male brain" theory posits that autism represents an extreme of the male pattern (impaired empathizing and enhanced systemizing). Here we suggest that specific aspects of autistic neuroanatomy may also be extremes of typical male neuroanatomy. PMID- 16272116 TI - Synthesis and structure of sila-adamantane. AB - An organosilicon cage compound containing the sila-adamantane structural unit, which is the building block of crystalline silicon, was synthesized by the Lewis acid-catalyzed rearrangement reaction of a structural isomer. PMID- 16272117 TI - Structures of the bacterial ribosome at 3.5 A resolution. AB - We describe two structures of the intact bacterial ribosome from Escherichia coli determined to a resolution of 3.5 angstroms by x-ray crystallography. These structures provide a detailed view of the interface between the small and large ribosomal subunits and the conformation of the peptidyl transferase center in the context of the intact ribosome. Differences between the two ribosomes reveal a high degree of flexibility between the head and the rest of the small subunit. Swiveling of the head of the small subunit observed in the present structures, coupled to the ratchet-like motion of the two subunits observed previously, suggests a mechanism for the final movements of messenger RNA (mRNA) and transfer RNAs (tRNAs) during translocation. PMID- 16272119 TI - Meteorite phosphates show constant 176Lu decay rate since 4557 million years ago. AB - The use of radioactive decay of 176Lu to 176Hf to study the evolution of the Earth requires a precise and accurate value for the 176Lu decay constant. Recent determinations of this decay constant by age comparison to the more precisely calibrated U-Pb isotopic system produced internally consistent but discrepant values between terrestrial minerals and meteorites. New highly radiogenic Lu-Hf data for phosphate minerals from Richardton (ordinary chondrite) and Acapulco (primitive achondrite) yield decay constant values of 1.864 x 10(-11) +/- 0.016 x 10(-11) and 1.832 x 10(-11) +/- 0.029 x 10(-11) year(-1), respectively, identical to the value determined from terrestrial minerals. PMID- 16272118 TI - A sporadic third layer in the ionosphere of Mars. AB - The daytime martian ionosphere has been observed as a two-layer structure with electron densities that peak at altitudes between about 110 and 130 kilometers. The Mars Express Orbiter Radio Science Experiment on the European Mars Express spacecraft observed, in 10 out of 120 electron density profiles, a third ionospheric layer at altitude ranges of 65 to 110 kilometers, where electron densities, on average, peaked at 0.8 x 10(10) per cubic meter. Such a layer has been predicted to be permanent and continuous. Its origin has been attributed to ablation of meteors and charge exchange of magnesium and iron. Our observations imply that this layer is present sporadically and locally. PMID- 16272120 TI - A direct role for dual oxidase in Drosophila gut immunity. AB - Because the mucosal epithelia are in constant contact with large numbers of microorganisms, these surfaces must be armed with efficient microbial control systems. Here, we show that the Drosophila nicotinamide adenine dinucleotide phosphate (NADPH) oxidase enzyme, dual oxidase (dDuox), is indispensable for gut antimicrobial activities. Adult flies in which dDuox expression is silenced showed a marked increase in mortality rate even after a minor infection through ingestion of microbe-contaminated food. This could be restored by the specific reintroduction of dDuox, demonstrating that this oxidase generates a unique epithelial oxidative burst that limits microbial proliferation in the gut. Thus, oxidant-mediated antimicrobial responses are not restricted to the phagocytes, but rather are used more broadly, including in mucosal barrier epithelia. PMID- 16272121 TI - Treatment of autoimmune neuroinflammation with a synthetic tryptophan metabolite. AB - Local catabolism of the amino acid tryptophan (Trp) by indoleamine 2,3 dioxygenase (IDO) is considered an important mechanism of regulating T cell immunity. We show that IDO transcription was increased when myelin-specific T cells were stimulated with tolerogenic altered self-peptides. Catabolites of Trp suppressed proliferation of myelin-specific T cells and inhibited production of proinflammatory T helper-1 (T(H)1) cytokines. N-(3,4,-Dimethoxycinnamoyl) anthranilic acid (3,4-DAA), an orally active synthetic derivative of the Trp metabolite anthranilic acid, reversed paralysis in mice with experimental autoimmune encephalomyelitis, a model of multiple sclerosis (MS). Trp catabolites and their derivatives offer a new strategy for treating T(H)1-mediated autoimmune diseases such as MS. PMID- 16272122 TI - Regulated pole-to-pole oscillations of a bacterial gliding motility protein. AB - Little is known about directed motility of bacteria that move by type IV pilus mediated (twitching) motility. Here, we found that during periodic cell reversals of Myxoccocus xanthus, type IV pili were disassembled at one pole and reassembled at the other pole. Accompanying these reversals, FrzS, a protein required for directed motility, moved in an oscillatory pattern between the cell poles. The frequency of the oscillations was controlled by the Frz chemosensory system, which is essential for directed motility. Pole-to-pole migration of FrzS appeared to involve movement along a filament running the length of the cell. FrzS dynamics may thus regulate cell polarity during directed motility. PMID- 16272123 TI - Retrograde signaling by Syt 4 induces presynaptic release and synapse-specific growth. AB - The molecular pathways involved in retrograde signal transduction at synapses and the function of retrograde communication are poorly understood. Here, we demonstrate that postsynaptic calcium 2+ ion (Ca2+) influx through glutamate receptors and subsequent postsynaptic vesicle fusion trigger a robust induction of presynaptic miniature release after high-frequency stimulation at Drosophila neuromuscular junctions. An isoform of the synaptotagmin family, synaptotagmin 4 (Syt 4), serves as a postsynaptic Ca2+ sensor to release retrograde signals that stimulate enhanced presynaptic function through activation of the cyclic adenosine monophosphate (cAMP)-cAMP-dependent protein kinase pathway. Postsynaptic Ca2+ influx also stimulates local synaptic differentiation and growth through Syt 4-mediated retrograde signals in a synapse-specific manner. PMID- 16272124 TI - Fast readout of object identity from macaque inferior temporal cortex. AB - Understanding the brain computations leading to object recognition requires quantitative characterization of the information represented in inferior temporal (IT) cortex. We used a biologically plausible, classifier-based readout technique to investigate the neural coding of selectivity and invariance at the IT population level. The activity of small neuronal populations (approximately 100 randomly selected cells) over very short time intervals (as small as 12.5 milliseconds) contained unexpectedly accurate and robust information about both object "identity" and "category." This information generalized over a range of object positions and scales, even for novel objects. Coarse information about position and scale could also be read out from the same population. PMID- 16272125 TI - Neuronal activity regulates diffusion across the neck of dendritic spines. AB - In mammalian excitatory neurons, dendritic spines are separated from dendrites by thin necks. Diffusion across the neck limits the chemical and electrical isolation of each spine. We found that spine/dendrite diffusional coupling is heterogeneous and uncovered a class of diffusionally isolated spines. The barrier to diffusion posed by the neck and the number of diffusionally isolated spines is bidirectionally regulated by neuronal activity. Furthermore, coincident synaptic activation and postsynaptic action potentials rapidly restrict diffusion across the neck. The regulation of diffusional coupling provides a possible mechanism for determining the amplitude of postsynaptic potentials and the accumulation of plasticity-inducing molecules within the spine head. PMID- 16272126 TI - Tissue-specific TAFs counteract Polycomb to turn on terminal differentiation. AB - Polycomb transcriptional silencing machinery is implicated in the maintenance of precursor fates, but how this repression is reversed to allow cell differentiation is unknown. Here we show that testis-specific TAF (TBP-associated factor) homologs required for terminal differentiation of male germ cells may activate target gene expression in part by counteracting repression by Polycomb. Chromatin immunoprecipitation revealed that testis TAFs bind to target promoters, reduce Polycomb binding, and promote local accumulation of H3K4me3, a mark of Trithorax action. Testis TAFs also promoted relocalization of Polycomb Repression Complex 1 components to the nucleolus in spermatocytes, implicating subnuclear architecture in the regulation of terminal differentiation. PMID- 16272127 TI - Influence of fluoro, chloro and alkyl alcohols on the folding pathway of human serum albumin. AB - Urea-induced equilibrium unfolding of human serum albumin (HSA) when studied by mean residue ellipticity at 222 nm (MRE(222)) or intrinsic fluorescence measurements showed a two-step, three-state transition with a stable intermediate around 4.6-5.2 M urea. The presence of 2,2,2-trifluoroethanol (TFE) resulted in a single-step, two-state transition with a significant shift towards higher urea concentration, suggesting the stabilizing effect of TFE. The free energy of stabilization (DeltaDeltaG(D)(H(2)O)) in the presence of 3.0 M TFE was determined to be 2.68 and 2.72 kcal/mol by MRE(222) and fluorescence measurements, respectively. The stabilizing potential of other alcohols on the refolding behavior of HSA at 5.0 M urea (where the intermediate exists) as studied by MRE(222) and intrinsic fluorescence measurements showed the following order: 1,1,1,3,3,3-hexafluoroisopropanol (HFIP) > TFE > 2-chloroethanol > tert-butanol > iso-propanol > ethanol > methanol. Further, the extent of refolding at the highest concentration of alcohol was similar in all cases. The stabilizing effect of TFE on guanidine hydrochloride (GdnHCl)-induced unfolding of HSA was nearly equal to that found for urea denaturation, as reflected in the DeltaDeltaG(D)(H(2)O) value (2.38 kcal/mol). Taken together, these results suggest that the stabilizing effect of TFE and other alcohols on urea/GdnHCl induced unfolding of HSA is higher for alcohols that contain bulky groups or fluorine atoms. PMID- 16272128 TI - Stabilization mechanism of the tryptophan synthase alpha-subunit from Thermus thermophilus HB8: X-ray crystallographic analysis and calorimetry. AB - In order to elucidate the thermo-stabilization mechanism of the tryptophan synthase alpha-subunit from the extreme thermophile Thermus thermophilus HB8 (Tt alpha-subunit), its crystal structure was determined and its stability was examined using DSC. The results were compared to those of other orthologs from mesophilic and hyperthermophilic organisms. The denaturation temperature of the Tt-alpha-subunit was higher than that of the alpha-subunit from S. typhimurium (St-alpha-subunit) but lower than that of the alpha-subunit from P. furiosus (Pf alpha-subunit). Specific denaturation enthalpy and specific denaturation heat capacity values of the Tt-alpha-subunit were the lowest among the three proteins, suggesting that entropy effects are responsible for the stabilization of the Tt alpha-subunit. Based on a structural comparison with the St-alpha-subunit, two deletions in loop regions, an increase in the number of ion pairs and a decrease in cavity volume seem to be responsible for the stabilization of the Tt-alpha subunit. The results of structural comparison suggest that the native structure of the Tt-alpha-subunit is better adapted to an ideally stable structure than that of the St-alpha-subunit, but worse than that of the Pf-alpha-subunit. The results of calorimetry suggest that the residual structure of the Tt-alpha subunit in the denatured state contributes to the stabilization. PMID- 16272129 TI - Catalytic properties of an organic solvent-resistant tyrosinase from Streptomyces sp. REN-21 and its high-level production in E. coli. AB - An organic solvent-resistant tyrosinase (OSRT) from Streptomyces sp. REN-21 is a unique enzyme showing high activity in the presence of organic solvents. The OSRT catalyzed oxidation of monophenols such as tyrosine-containing peptides and proteins was examined. The catalytic properties of OSRT were compared with those of mushroom tyrosinase. OSRT was shown to oxidize Gly-l-Tyr most effectively among four peptide substrates tested. On the other hand, mushroom tyrosinase showed the highest activity toward l-Tyr-Gly under the condition of 1 mM substrate. OSRT oxidized several proteins, including casein and hemoglobin, with relatively higher activity compared with mushroom tyrosinase under the condition of 1% (w/v) substrate. Thus, it was clarified that the catalytic properties of OSRT toward tyrosine-containing peptides and proteins are different from those of mushroom tyrosinase under these conditions. The OSRT-encoding gene operon was cloned, and found to consist of two genes, designated ORF-OSRT and ORF-393. The former encodes apo-OSRT, and the latter encodes the putative activator protein of apo-OSRT. A binuclear copper-binding site (type-3 copper site) characteristic of tyrosinases is contained in the deduced amino acid sequence for apo-OSRT. A high level production system for the OSRT was constructed using pET20b(+) and Escherichia coli BL21(DE3)pLysS. Approximately 54 mg of active OSRT was synthesized in a 1-liter broth culture by this system. The properties of the recombinant OSRT were similar to those of the wild-type enzyme. In conclusion, we succeeded in constructing a high-level production system for OSRT. PMID- 16272131 TI - Genes specifically expressed in sexually differentiated female spheroids of Volvox carteri. AB - Volvox carteri is a multicellular green alga with only two cell types, somatic cells and reproductive cells. Phylogenetic analysis suggests that this organism has evolved from a Chlamydomonas-like unicellular ancestor along with multicellularity, cellular differentiation, and a change in the mode of sexual reproduction from isogamy to oogamy. To examine the mechanism of sexual differentiation and the evolution of oogamy, we isolated 6 different cDNA sequences specifically expressed in sexually differentiated female spheroids. The genes for the cDNAs were designated SEF1 to SEF6. The time course of accumulation of each mRNA was shown to be distinct. The expression of some of these genes was not significantly affected when the sexual inducer was removed after the induction of sexual development. Sequence analysis indicates that SEF5 and SEF6 encode pherophorin-related proteins. Of these, SEF5 has the unique structural feature of a polyproline stretch in the C-terminal domain in addition to the one found in the central region. PMID- 16272130 TI - Single-strand conformation polymorphism (SSCP) of oligodeoxyribonucleotides: an insight into solution structural dynamics of DNAs provided by gel electrophoresis and molecular dynamics simulations. AB - Studies on the solution structure dynamics of RNA/DNA are becoming crucially important. The phenomena of SSCP (single-strand conformation polymorphism), small RNA dynamics in a cell, and others can be related to the conformational changes of single-stranded (ss) RNAs/DNAs in solution. However, little is known about those dynamics. Only the intra-structural transition of ssDNAs in solution has been reported based on Watson-Crick (W-C) base-pairing. Here, we found a general feature of the SSCP phenomenon by studying the simpler molecules of ss oligodeoxyribonucleotides. A single base substitution or a positional exchange of nucleotide in a highly homologous series of ss-dodecanucleotides led to a change in the mobility-in-gel. This was unexpected, since most of these nucleotides [such as d(A(11)G) or d(A(11)C)] have no possibility of forming W-C base-pairing. MD (molecular dynamics) experiments revealed differences in shape and size between the dynamic structures of these molecules which could affect their mobility-in-gel. In addition, a high correlation was observed between the electrophoretic mobility and the size-related parameters such as end-to-end distance obtained from MD simulations. Because the simulation was considerably shorter (nanosecond) than the experimental time-scale (second), the result must be considered conservatively; but it is nevertheless encouraging for utilizing MD simulation for structural analysis of oligonucleotides. PMID- 16272132 TI - Complete substitutional analysis of a sunflower trypsin inhibitor with different serine proteases. AB - Here we present a method to simultaneously characterize and/or optimize both the binding loop towards the protease and a cysteine-stabilized scaffold. The small peptidic sunflower trypsin inhibitor (SFTI-1) was chosen as a model system for these experiments. The inhibitor was investigated for positional specificity against trypsin, elastase and proteinase K using complete substitutional analyses based on cellulose-bound peptide spot synthesis. Inhibitor variants optimized for elastase or proteinase K inhibition by several rounds of substitutional analyses exhibit K(i) values in the micromolar range and high specificity for the corresponding protease. The results of this easy-to-perform assay can be used to design an improved peptide library using classical methods. PMID- 16272133 TI - N-acetyltransferase Mpr1 confers freeze tolerance on Saccharomyces cerevisiae by reducing reactive oxygen species. AB - N-Acetyltransferase Mpr1 of Saccharomyces cerevisiae can reduce intracellular oxidation levels and protect yeast cells under oxidative stress. We found that yeast cells exhibited increased levels of reactive oxygen species during freezing and thawing. Gene disruption and expression experiments indicated that Mpr1 protects yeast cells from freezing stress by reducing the intracellular levels of reactive oxygen species. The combination of Mpr1 and l-proline could further enhance the resistance to freezing stress. Hence, Mpr1 as well as l-proline has promising potential for the breeding of novel freeze-tolerant yeast strains. PMID- 16272134 TI - Expression of adipose differentiation-related protein (ADRP) is conjointly regulated by PU.1 and AP-1 in macrophages. AB - ADRP is associated with intracellular lipid droplets. We demonstrate the regulatory mechanism for ADRP expression in RAW264.7 macrophages. The ADRP mRNA expression was stimulated by PMA, and synergistically enhanced in association with its protein level in the presence of lipids. A proteasome inhibitor protected the protein from degradation under the lipid-free conditions. One of the possible sites of the PMA action was proved to be an Ets/AP-1 element in the promoter, since mutations of this site reduced the PMA-induced promoter activity, and ligation of this element led to a significant increase in the PMA responsiveness of homologous or heterologous promoters. Mutations of this site diminished the synergistic effect on the promoter activity induced by PMA and oleic acid, suggesting a possible interaction between this site and the downstream PPARdelta site. EMSA revealed that PU.1 and AP-1 conjointly bound to this site. The juxtaposition of the two sequences was requisite for full activity, since spacer sequences between them decreased the PMA-induced activity. PI3 kinase inhibitor was found to reduce the PMA-induced mRNA expression and promoter activity in parallel with PU.1/AP-1 complex formation on EMSA. From these results, we concluded that the Ets/AP-1 site is an important cis-acting element that regulates the ADRP gene expression in macrophages. PMID- 16272135 TI - Possible role of each repeat structure of the microtubule-binding domain of the tau protein in in vitro aggregation. AB - Although one of the priorities in Alzheimer's research is to clarify the filament formation mechanism of the tau protein, it is currently unclear how it is transformed from a normal structure in a neuron. To examine which part and what structural change in the tau protein are involved in its transformation into a pathological entity, the initial in vitro self-aggregation features of each repeat peptide (R1-R4) constituting a three- or four-repeat microtubule-binding domain (3RMBD or 4RMBD) in the tau protein was investigated by measuring both the fluorescence and light scattering (LS) spectra on the same instrument, because these MBD domains constitute the core moiety of the tau paired helical filament (PHF) structure. The conformational features of the R1 and R4 peptides in trifluoroethanol were also investigated by (1)H-NMR and molecular modeling analyses and compared with those of the R2 and R3 peptides. The analyses of the LS spectra clarified (i) the self-aggregation rates of R1-R4, 3RMBD and 4RMBD at a fixed concentration (15 mM), (ii) their minimum concentrations for starting filament extension, and (iii) the concentration dependence of their self aggregations. The fluorescence analyses showed that the R2 and R3 peptides have high self-aggregation abilities at the extension and nucleation steps, respectively, in their filament formation processes. It was shown that the R2 repeat exhibits a positive synergistic effect on the aggregation of 4RMBD. The R1 and R4 repeats, despite their weak self-aggregation abilities, are necessary for the intact PHF formation of tau MBD, whereas they exerted a negative effect on the R3-driven aggregation of 3RMBD. The conformational analyses showed the importance of the amphipathic conformational features of the R1 to R4 peptides, and the intermolecular disulfide bonding abilities of the R2 and R3 peptides for the PHF formation. On the basis of the present spectral and conformational results, the possible role of each repeat structure in the dimeric formation of MBD at the initial in vitro aggregation stage is discussed. PMID- 16272136 TI - Topogenic properties of transmembrane segments of Arabidopsis thaliana NHX1 reveal a common topology model of the Na+/H+ exchanger family. AB - The membrane topology of the Arabidopsis thaliana Na(+)/H(+) exchanger isoform 1 (AtNHX1) was investigated by examining the topogenic function of transmembrane (TM) segments using a cell-free system. Even though the signal peptide found in the human Na(+)/H(+) exchanger (NHE) family is missing, the N-terminal hydrophobic segment was efficiently inserted into the membrane and had an N terminus lumen topology depending on the next TM segment. The two N-terminal TM segments had the same topology as those of TM2 and TM3 of human NHE1. In contrast, TM2 and TM3 of human NHE1 did not acquire the correct topology when the signal peptide (denoted as TM1) was deleted. Furthermore, there were three hydrophobic segments with the same topogenic properties as the TM9-H10-TM10 segments of human NHE1, which has one lumenal loop (H10) and two flanking TM segments (TM9 and TM10). These data indicate that the plant NHX isoforms can form the common membrane topology proposed for the human NHE family, even though it does not have a signal peptide. PMID- 16272138 TI - Structural insights into the asymmetric effects of zinc-ligand cysteine mutations in the novel zinc ribbon domain of human TFIIEalpha for transcription. AB - The large subunit of TFIIE (TFIIEalpha) has a highly conserved zinc ribbon domain, which is essential for transcription. Recently, we determined the solution structure of this domain to be that of a novel zinc finger motif [Okuda et al. (2004) J. Biol. Chem. 279, 51395-51403]. On examination of the functions of four cysteine mutants of TFIIEalpha, in which each of four zinc-liganded cysteines was replaced by alanine, we found an interesting functional asymmetry; on a supercoiled template, the two C-terminal mutants did not show any transcriptional activity, however, the two N-terminal mutants retained about 20% activity. Furthermore, these two pairs of mutants showed distinct binding abilities as to several general transcription factors. To obtain structural insights into the asymmetry, here we have analyzed the structures of the four cysteine mutants of the zinc ribbon domain by CD and NMR. All four mutants possessed a characteristic partially folded structure coordinating with a zinc atom, despite the imperfect set of cysteine-ligands. However, they equilibrated with several structures including the random coil structure. Unexpectedly, the two N-terminal mutants mainly equilibrated with the random coil structure, while the two C-terminal ones mainly equilibrated with folded structures. The characteristic structure formation of each mutant was reversible, which totally depended on the zinc binding. PMID- 16272137 TI - Human anti-human IL-18 antibody recognizing the IL-18-binding site 3 with IL-18 signaling blocking activity. AB - IL-18 is an important regulator in both innate and acquired immune responses. The aberrant expression of IL-18 is associated with severe inflammatory conditions, such as autoimmune diseases and allergies. Thus, human antibodies with inhibitory activity on IL-18 signaling may be useful for therapeutic applications. We report here the first establishment of an antagonistic anti-IL-18 complete human antibody, h18-108, employing a human single chain antibody (scFv)-displaying phage library. The h18-108 scFv inhibited the IFN-gamma production of a human myelomonocytic cell line, KG-1. Flow cytometry analysis showed that h18-108 blocked the binding of IL-18 to KG-1 cells. Epitope mapping analysis using two kinds of random peptide-displaying phage libraries and an IL-18 alanine mutant (D98A) demonstrated that the h18-108 scFv binds to the site 3 of IL-18, which is suggested to be an association site with the IL-18 receptor beta. The complete human Fab and IgG forms of h18-108 have been successfully constructed to attain increases in both binding affinity and inhibitory activity. PMID- 16272139 TI - Phospholipase Cdelta4 associates with glutamate receptor interacting protein 1 in testis. AB - We reported previously that phospholipase C (PLC) delta4 is required for calcium mobilization in the zona pellucida-induced acrosome reaction in sperm. Here we focused on the function of the C2 domain of PLCdelta4 and report that glutamate receptor-interacting protein1 (GRIP1) was identified as a binding protein of the PLCdelta4-C2 domain on yeast two-hybrid screening. Physiological interaction of GRIP1 with PLCdelta4 in mouse testis was confirmed by immunoprecipitation with anti-PLCdelta4 antibodies and the association seemed to correlate with the maturation stage of sperm. We also determined that a PDZ-binding motif at the C terminus of the PLCdelta4-C2 domain is responsible for GRIP1 binding, whereas the sixth or seventh PDZ domain of GRIP1 is essential and sufficient for association with the PLCdelta4-C2 domain. These results indicate that PLCdelta4 binds via its C2 domain to the PDZ6 or PDZ7 domain of GRIP1, and that this association may play a role in spermatogenesis. PMID- 16272140 TI - Repression of GR-mediated expression of the tryptophan oxygenase gene by the SWI/SNF complex during liver development. AB - The chromatin remodeling complex, SWI/SNF, is known to regulate the transcription of several genes by altering the chromatin structure in an ATP-dependent manner. SWI/SNF exclusively contains BRG1 or BRM as an ATPase subunit. In the present study, we studied the role of SWI/SNF containing BRM or BRG1 in the expression of the liver-specific tryptophan oxygenase (TO) and tyrosine aminotransferase genes. Chromatin remodeling factors significantly repressed the expression of these genes induced by glucocorticoid receptor and dexamethasone. Since the repression was not reversed by trichostatin A treatment, it seemed to be independent of the well-known histone deacetylase pathway. Knock-down of BRG1 by small interfering RNA reversed the repression in primary fetal hepatocytes. These results support a model in which SWI/SNF containing BRG1 represses late stage-specific TO gene expression at an early stage of liver development. PMID- 16272141 TI - Remodeling of actin cytoskeleton in lupeol-induced B16 2F2 cell differentiation. AB - Lupeol induces the formation of dendrites in B16 2F2 melanoma cells. The remodeling of cytoskeletal components contributes to the dendricity of melanoma cells. We studied the effects of lupeol on the remodeling of cytoplasmic filaments in B16 2F2 cells. Western blotting revealed no change in the levels of actin and tubulin. Lupeol attenuated stress fiber assembly, but did not promote the remodeling of microtubular networks. We examined the activation of cofilin, an actin-depolymerizing factor, in lupeol-treated B16 2F2 cells by western blotting. The level of phospho-cofilin was found to decrease in a time-dependent manner. Inhibition of p38 MAPK by SB203580 blocked tyrosinase induction by lupeol, but did not influence the disruption of stress fiber assembly or the dephosphorylation of cofilin. Furthermore, we studied the effects of lupeol on cell migration. At 10 microM, lupeol markedly inhibited the haptotaxis of B16 2F2 cells to fibronectin. Additionally, lupeol strongly inhibited the migration of human melanoma and neuroblastoma cells, and weakly suppressed the migration of lung adenocarcinoma cells. However, lupeol did not affect the motility of other cancer cells. The results suggest that lupeol suppresses the migration of malignant melanoma cells by disassembling the actin cytoskeleton. PMID- 16272143 TI - Interaction of N1,N12-diacetylspermine with polyamine transport systems of polarized porcine renal cell line LLC-PK1. AB - LLC-PK(1) cells grown on porous membrane filters were employed as a model system to explore the renal transport of polyamines. The polarity of LLC-PK(1) monolayers was confirmed by the exclusive appearance of a Na(+)-dependent alpha methylglucoside transport system on the apical surface. The uptake of free polyamines from the basolateral side of monolayers was consistent with the existence of a single class of transport system, while the existence of two kinetically distinct polyamine transport systems with higher and lower affinities on apical membranes was suggested. The results of competition studies indicated that each of these transporters was able to interact with putrescine, spermidine and spermine. LLC-PK(1) cells incorporated monoacetylspermine from the apical surface of monolayers at about half the rate of spermine uptake. Monoacetylspermine inhibited spermidine uptake, indicating that free polyamine transport systems also recognized the monoacetylated derivative. In contrast, N(1),N(12)-diacetylspermine did not inhibit spermidine uptake, nor was it incorporated into the cells, indicating the absence of transport systems that recognize N(1),N(12)-diacetylspermine on the apical membranes of LLC-PK(1) cells. These results may be relevant as to our previous observation that the content of diacetylpolyamines in urine is relatively constant, and may explain the excellence of N(1),N(12)-diacetylspermine as a tumor marker. PMID- 16272142 TI - Exo-Taq-based detection of DNA-binding protein for homogeneous and microarray format. AB - The study of DNA-protein interactions is of great importance to understand basic cellular processes such as transcription, replication and recombination. In this research, we developed a novel detection system for DNA-binding proteins (DBPs) involving the exonuclease (Exo) III and Taq DNA polymerase reactions. The system consists of three steps, as follows: the target DBP in the sample solution is incubated with probe DNA, and the probe is digested with Exo III and then extended with Taq using fluorescent dye-labeled dUTP as a substrate. The DBP protects the probe from digestion by Exo III. Therefore, only the DBP-bound probe allows the following extension. We examined this system using the lambda phage Cro repressor in a homogeneous format. The fluorescence image after gel electrophoresis showed a specific band. We also found that this system could be applied to the rapid and efficient detection of DBPs in stem and loop ds-DNA array formats. These results suggest that our method is useful as a new tool for analyzing DNA-protein interactions. PMID- 16272144 TI - Regulation of intracellular localization and transcriptional activity of FOXO4 by protein kinase B through phosphorylation at the motif sites conserved among the FOXO family. AB - FOXO4 transcription factor, also referred to AFX, contains three putative phosphorylation motif sites for protein kinase B (PKB), Thr32, Ser197, and Ser262, and it is proposed that phosphorylated FOXO4 stays in the cytosol and is imported to the nucleus through dephosphorylation to induce target gene expression. These three sites were revealed to be phosphorylated by PKB in vitro on phosphopeptide analysis, and in cultured cells on immunoblotting with phosphorylation-site specific antibodies. The mutants with either Thr32 or Ser197 replaced by Ala were found mostly in the nuclear but not the cytosol fraction, and treatment with platelet-derived growth factor did not change their distributions in the cells. FOXO4 proteins mutated at these two sites showed 3- to 5-fold higher transcriptional activity than that of the wild type. In contrast, the replacement of Ser262 did not alter the localization or transcriptional activity. These results indicate that phosphorylation at Thr32 and Ser197 is indispensable, whereas that at Ser262 is not critical, for regulation of the nuclear localization and transcriptional activity of FOXO4. These properties are similar to those of FOXO1 and FOXO3, and thus FOXO transcription factors seem to be regulated through a common mechanism by PKB in the growth factor signaling pathway. PMID- 16272145 TI - Formation of retinoylated proteins from retinoyl-CoA in rat tissues. AB - Retinoylation (acylation of proteins by retinoic acid) is considered as one mechanism of retinoic acid (RA) action occurring in cells in vitro and in vivo. Previously, our studies showed that in rat tissues the formation of retinoyl-CoA from RA, the first step of retinoylation, required ATP, CoA and MgCl(2). In the current study, we examined whether the transfer of retinoyl-CoA into proteins, the second step of retinoylation, occurs in rat tissues. [(3)H]-Labeled-retinoyl CoA bound covalently to proteins in rat liver, kidney, testis, and brain. The levels of incorporation of retinoyl-CoA into proteins were higher in vitamin A deficient rats than in normal ones. The formation of retinoylated proteins depended on the incubation time, and the concentrations of retinoyl-CoA and homogenate. The reaction was suppressed by fatty acyl-CoAs and palmitic acid, but not by arachidonic acid. The Vmax and Km values for retinoyl-CoA in the formation of retinoylated proteins using a crude liver extract were estimated to be 2,597.3 pmol/min/mg protein and 9.5 x 10(-5) M, respectively. Retinoylated proteins formed from retinoyl-CoA, including a 17 kDa protein exhibiting high radioactivity, disappeared in the presence of 2-mercaptoethanol, indicating that RA was linked to the proteins through a thioester bond. These results demonstrate that retinoylation in rat tissues occurs via retinoyl-CoA formed from RA. This process may play a significant physiological role in cells. PMID- 16272146 TI - The ER stress pathway involving CHOP is activated in the lungs of LPS-treated mice. AB - CHOP is a C/EBP family transcription factor involved in endoplasmic reticulum (ER) stress-mediated apoptosis. To determine if the ER stress pathway is involved in the pathogenesis of LPS-treated mouse lung injury, mice were given lipopolysaccharide (LPS) intraperitoneally. The mRNAs for activating transcription factor (ATF) 4 and X-box binding protein (XBP) 1, transcriptional activators of the CHOP gene, and that for CHOP were induced by or after the LPS treatment. Apoptosis induced by LPS treatment was suppressed in the lungs of Chop knockout mice. Overexpression of CHOP induced apoptosis in a lung cancer-derived cell line. These results suggest that the ER stress pathway, involving CHOP, is activated and plays a role in the pathogenesis of septic shock lung. PMID- 16272148 TI - Two mouse cofilin isoforms, muscle-type (MCF) and non-muscle type (NMCF), interact with F-actin with different efficiencies. AB - Two cofilin isoforms, a muscle-type (MCF) and a non-muscle-type (NMCF), are co expressed in developing mammalian skeletal and cardiac muscles. To clarify how they are involved in the actin filament dynamics during myofibrillogenesis, we examined their localization in muscle tissues and cultured muscle cells using immunocytochemical methods, and their interaction with F-actin in vitro. NMCF was mostly detected in a diffuse pattern in the cytoplasm but MCF was partly localized to the striated structures in myofibrils. The location of chicken cofilin, a homologue of MCF, in the I-bands of myofibrils was determined by an immunocytochemical method. It is suggested that MCF could be associated with actin filaments in muscle cells more efficiently than NMCF. Using purified recombinant MCF and NMCF, their interaction with F-actin was examined in vitro by a cosedimentation assay method. We observed that MCF was precipitated with F actin more effectively than NMCF. When MCF and NMCF were simultaneously incubated with F-actin, MCF was preferentially associated with F-actin. MCF and NMCF inhibited the interaction of F-actin with tropomyosin, but the former suppressed the actin-tropomyosin interaction more strongly than the latter. These results suggest that MCF interacts with F-actin with higher affinity than NMCF, and although both of them are involved in the regulation of actin assembly in developing myotubes, the two proteins may play somewhat different roles. PMID- 16272147 TI - Red blood cells highly express type I platelet-activating factor-acetylhydrolase (PAF-AH) which consists of the alpha1/alpha2 complex. AB - Although red blood cells account for about 30% of total PAF-AH activity found in the blood, the physiological function of this enzyme is unknown. To understand the role and regulatory mechanism of this enzyme, we purified it from easily obtainable pig red blood cells. PAF-AH activity was mainly found in the soluble fraction of the red blood cells. Two peaks of enzyme activity appeared with increasing concentration of imidazole on column chromatography on nickel nitroacetic acid (Ni-NTA) resin. We called these peaks of small and large enzyme activities fractions X and Y, respectively, and then further purified the enzymes by sequential chromatofocusing on Mono P and gel filtration on TSK G-3000. In the final preparation from fraction Y, two proteins bands corresponding to 26 kDa and 28 kDa were related to enzyme activity. Determination of the partial amino acid sequences of the proteins of 26 kDa and 28 kDa revealed that these proteins were identical to alpha(1) and alpha(2), respectively, both of which are catalytic subunits of Type I intracellular PAF-AH. On Western analysis, the 26 kDa and 28 kDa protein bands cross-reacted with specific monoclonal antibodies to alpha(1) and alpha(2), respectively. Since the apparent molecular weight of the natural enzyme was estimated to be about 60 kDa, the enzyme activity in fraction Y was thought to be that of a heterodimer consisting of alpha(1) and alpha(2). On the other hand, the enzyme activity in fraction X was thought to be that of a homodimer consisting of alpha(2). Other blood cells such as polymorphonuclear leukocytes and platelets only contained the alpha(2)/alpha(2) homodimer. It has been reported that the alpha(1)/alpha(2) heterodimer is poorly expressed in adult animals except for in the spermatogonium. Taken altogether, these results suggest that high expression of the alpha(1)/alpha(2) heterodimer is important for the physiological function of mature red blood cells. PMID- 16272149 TI - Starvation-induced cleavage of the tRNA anticodon loop in Tetrahymena thermophila. AB - Amino acid deprivation triggers dramatic physiological responses in all organisms, altering both the synthesis and destruction of RNA and protein. Here we describe, using the ciliate Tetrahymena thermophila, a previously unidentified response to amino acid deprivation in which mature transfer RNA (tRNA) is cleaved in the anticodon loop. We observed that anticodon loop cleavage affects a small fraction of most or all tRNA sequences. Accumulation of cleaved tRNA is temporally coordinated with the morphological and metabolic changes of adaptation to starvation. The starvation-induced endonucleolytic cleavage activity targets tRNAs that have undergone maturation by 5' and 3' end processing and base modification. Curiously, the majority of cleaved tRNAs lack the 3' terminal CCA nucleotides required for aminoacylation. Starvation-induced tRNA cleavage is inhibited in the presence of essential amino acids, independent of the persistence of other starvation-induced responses. Our findings suggest that anticodon loop cleavage may reduce the accumulation of uncharged tRNAs as part of a specific response induced by amino acid starvation. PMID- 16272150 TI - Biochemical properties of human pantothenate kinase 2 isoforms and mutations linked to pantothenate kinase-associated neurodegeneration. AB - The PANK2 gene encodes the human pantothenate kinase 2 protein isoforms, and PANK2 mutations are linked to pantothenate kinase-associated neurodegeneration. Two PanK2 protein forms are proteolytically processed to form a mitochondrially localized, mature PanK2. Another isoform arose from a proposed initiation at a leucine codon and was not processed further. The fifth isoform was postulated to arise from an alternative splicing event and was found to encode an inactive protein. Fourteen mutant PanK2 proteins with single amino acid substitutions, associated with either early or late onset disease, were evaluated for activity. The PanK2(G521R), the most frequent mutation in pantothenate kinase-associated neurodegeneration, was devoid of activity and did not fold properly. However, nine of the mutant proteins associated with disease possessed catalytic activities that were indistinguishable from wild type, including the frequently encountered PanK2(T528M) missense mutation. PanK2 was extremely sensitive to feedback inhibition by CoA thioesters (IC50 values between 250 and 500 nM), and the regulation of the active PanK2 mutants was comparable with that of the wild type protein. Coexpression of the PanK2(G521R) and wild-type PanK2 did not interfere with wild-type enzyme activity, arguing against a dominant negative effect of the PanK2(G521R) mutation in heterozygous patients. These data described the unique biochemical features of the PanK2 isoforms and suggested that catalytic defects may not be the sole cause for the neurodegenerative phenotype. PMID- 16272151 TI - p38 Mitogen-activated protein kinase plays a stimulatory role in hepatic gluconeogenesis. AB - Hepatic gluconeogenesis is essential for maintaining blood glucose levels during fasting and is the major contributor to postprandial and fasting hyperglycemia in diabetes. Gluconeogenesis is a classic cAMP/protein kinase A-dependent process initiated by glucagon, which is elevated in the blood during fasting and in diabetes. In this study, we have shown that p38 mitogen-activated protein kinase (p38) was activated in liver by fasting and in primary hepatocytes by glucagon or forskolin. Fasting plasma glucose levels were reduced upon blockade of p38 with either a chemical inhibitor or small interference RNA in mice. In examining the mechanism, inhibition of p38 suppressed gluconeogenesis in liver, along with expression of key gluconeogenic genes, including phosphoenolpyruvate carboxykinase and glucose-6-phosphatase. Peroxisome proliferator-activated receptor gamma coactivator 1alpha and cAMP-response element-binding protein have been shown to be important mediators of hepatic gluconeogenesis. We have shown that inhibition of p38 prevented transcription of the PPARgamma coactivator 1alpha gene as well as phosphorylation of cAMP-response element-binding protein. Together, our results from in vitro and in vivo studies define a model in which cAMP-dependent activation of genes involved in gluconeogenesis is dependent upon the p38 pathway, thus adding a new player to our evolving understanding of this physiology. PMID- 16272152 TI - Tyr-95 and Ile-172 in transmembrane segments 1 and 3 of human serotonin transporters interact to establish high affinity recognition of antidepressants. AB - In previous studies examining the structural determinants of antidepressant and substrate recognition by serotonin transporters (SERTs), we identified Tyr-95 in transmembrane segment 1 (TM1) of human SERT as a major determinant of binding for several antagonists, including racemic citalopram ((RS)-CIT). Here we described a separate site in hSERT TM3 (Ile-172) that impacts (RS)-CIT recognition when switched to the corresponding Drosophila SERT residue (I172M). The hSERT I172M mutant displays a marked loss of inhibitor potency for multiple inhibitors such as (RS)-CIT, clomipramine, RTI-55, fluoxetine, cocaine, nisoxetine, mazindol, and nomifensine, whereas recognition of substrates, including serotonin and 3,4 methylenedioxymethamphetamine, is unaffected. Selectivity for antagonist interactions is evident with this substitution because the potencies of the antidepressants tianeptine and paroxetine are unchanged. Reduced cocaine analog recognition was verified in photoaffinity labeling studies using [(125)I]MFZ 2 24. In contrast to the I172M substitution, other substitutions at this position significantly affected substrate recognition and/or transport activity. Additionally, the mouse mutation (mSERT I172M) exhibits similar selective changes in inhibitor potency. Unlike hSERT or mSERT, analogous substitutions in mouse dopamine transporter (V152M) or human norepinephrine transporter (V148M) result in transporters that bind substrate but are deficient in the subsequent translocation of the substrate. A double mutant hSERT Y95F/I172M had a synergistic impact on (RS)-CIT recognition ( approximately 10,000-fold decrease in (RS)-CIT potency) in the context of normal serotonin recognition. The less active enantiomer (R)-CIT responded to the I172M substitution like (S)-CIT but was relatively insensitive to the Y95F substitution and did not display a synergistic loss at Y95F/I172M. An hSERT mutant with single cysteine substitutions in TM1 and TM3 resulted in formation of a high affinity cadmium metal coordination site, suggesting proximity of these domains in the tertiary structure of SERT. These studies provided evidence for distinct binding sites coordinating SERT antagonists and revealed a close interaction between TM1 and TM3 differentially targeted by stereoisomers of CIT. PMID- 16272153 TI - Extrasynaptic membrane trafficking regulated by GluR1 serine 845 phosphorylation primes AMPA receptors for long-term potentiation. AB - Enhancement of synaptic transmission, as occurs in long-term potentiation (LTP), can result from several mechanisms that are regulated by phosphorylation of the AMPA-type glutamate receptor (AMPAR). Using a quantitative assay of net serine 845 (Ser-845) phosphorylation in the GluR1 subunit of AMPARs, we investigated the relationship between phospho-Ser-845, GluR1 surface expression, and synaptic strength in hippocampal neurons. About 15% of surface AMPARs in cultured neurons were phosphorylated at Ser-845 basally, whereas chemical potentiation (forskolin/rolipram treatment) persistently increased this to 60% and chemical depression (N-methyl-D-aspartate treatment) decreased it to 10%. These changes in Ser-845 phosphorylation were paralleled by corresponding changes in the surface expression of AMPARs in both cultured neurons and hippocampal slices. For every 1% increase in net phospho-Ser-845, there was 0.75% increase in the surface fraction of GluR1. Phosphorylation of Ser-845 correlated with a selective delivery of AMPARs to extrasynaptic sites, and their synaptic localization required coincident synaptic activity. Furthermore, increasing the extrasynaptic pool of AMPA receptors resulted in stronger theta burst LTP. Our results support a two-step model for delivery of GluR1-containing AMPARs to synapses during activity-dependent LTP, where Ser-845 phosphorylation can traffic AMPARs to extrasynaptic sites for subsequent delivery to synapses during LTP. PMID- 16272154 TI - Crystal structure of an archaeal pentameric riboflavin synthase in complex with a substrate analog inhibitor: stereochemical implications. AB - Whereas eubacterial and eukaryotic riboflavin synthases form homotrimers, archaeal riboflavin synthases from Methanocaldococcus jannaschii and Methanothermobacter thermoautrophicus are homopentamers with sequence similarity to the 6,7-dimethyl-8-ribityllumazine synthase catalyzing the penultimate step in riboflavin biosynthesis. Recently it could be shown that the complex dismutation reaction catalyzed by the pentameric M. jannaschii riboflavin synthase generates riboflavin with the same regiochemistry as observed for trimeric riboflavin synthases. Here we present crystal structures of the pentameric riboflavin synthase from M. jannaschii and its complex with the substrate analog inhibitor, 6,7-dioxo-8-ribityllumazine. The complex structure shows five active sites located between adjacent monomers of the pentamer. Each active site can accommodate two substrate analog molecules in anti-parallel orientation. The topology of the two bound ligands at the active site is well in line with the known stereochemistry of a pentacyclic adduct of 6,7-dimethyl-8-ribityllumazine that has been shown to serve as a kinetically competent intermediate. The pentacyclic intermediates of trimeric and pentameric riboflavin synthases are diastereomers. PMID- 16272155 TI - Mdv1 interacts with assembled dnm1 to promote mitochondrial division. AB - The dynamin-related GTPase, Dnm1, self-assembles into punctate structures that are targeted to the outer mitochondrial membrane where they mediate mitochondrial division. Post-targeting, Dnm1-dependent division is controlled by the actions of the WD repeat protein, Mdv1, and the mitochondrial tetratricopeptide repeat-like outer membrane protein, Fis1. Our previous studies suggest a model where at this step Mdv1 functions as an adaptor linking Fis1 with Dnm1. To gain insight into the exact role of the Fis1.Mdv1.Dnm1 complex in mitochondrial division, we performed a structure-function analysis of the Mdv1 adaptor. Our analysis suggests that dynamic interactions between Mdv1 and Dnm1 play a key role in division by regulating Dnm1 self-assembly. PMID- 16272156 TI - The guanine nucleotide exchange factor CNrasGEF regulates melanogenesis and cell survival in melanoma cells. AB - cAMP-dependent Ras activation has been demonstrated in numerous cell types, particularly of neuronal (including melanoma cells) and endocrine origin, but the Ras activator involved has not been identified. In B16 melanoma cells, cAMP activates the Ras/Erk pathway, leading initially to stimulation but subsequently to long term (>24-h) inhibition of melanogenesis (dendrite extension and melanin production). Here we identify CNrasGEF as the Ras guanine nucleotide exchange factor (GEF) involved. We demonstrate that CNrasGEF is expressed endogenously in B16 melanoma cells and that cAMP-mediated activation of Ras and Erk1/2 in these cells can be augmented by CNrasGEF overexpression and reduced by its knockdown by RNA interference. Moreover, we show that CNrasGEF participates in the regulation of melanogenesis. Knockdown of CNrasGEF leads to increased dendrite extension and melanin production observed approximately 50 h after forskolin/isobutylmethylxanthine treatment, suggesting that CNrasGEF inhibits melanogenesis in the long term. Independently, we find that overexpression of CNrasGEF leads to apoptosis, whereas its knockdown by RNAi enhances cell proliferation, independent of cAMP. Collectively, these results suggest that CNrasGEF regulates melanogenesis but that it also has a distinct role in regulating cell proliferation/apoptosis. PMID- 16272157 TI - Phosphatidylinositol 3-kinase activity is critical for glucose metabolism and embryo survival in murine blastocysts. AB - The phosphatidylinositol 3-kinase (PI3K) signal transduction pathway is a well known mediator of cell growth, proliferation, and survival signals. Whereas the expression and function of this pathway has been documented during mammalian development, evidence demonstrating the physiologic importance of this pathway in murine preimplantation embryos is beginning to emerge. This study demonstrates that inhibition of the PI3K pathway leads to the induction of apoptosis in both murine blastocysts and trophoblast stem cells. The apoptosis induced in both model systems correlates with a decrease in the expression of the glucose transporter GLUT1 at the plasma membrane. In addition, blastocysts cultured in the presence of the PI3K inhibitor LY-294002 display a decrease in both 2 deoxyglucose uptake and hexokinase activity as compared with control blastocysts. To determine the impact of PI3K inhibition on pregnancy outcome, embryo transfer experiments were performed. Blastocysts cultured in the presence of LY-294002 demonstrate a dramatic increase in fetal resorptions as compared with control embryos. Finally, we demonstrate that impairment of glucose metabolism via iodoacetate, a glyceraldehyde-3-phosphate dehydrogenase inhibitor, is sufficient to induce apoptosis in both blastocysts and trophoblast stem cells. Moreover, blastocysts treated with iodoacetate result in poor pregnancy outcome as determined by embryo transfer experiments. Taken together these data demonstrate the critical importance of the PI3K pathway in preimplantation embryo survival and pregnancy outcome and further emphasize the importance of glucose utilization and metabolism in cell survival pathways. PMID- 16272158 TI - Binding of PAI-1 to endothelial cells stimulated by thymosin beta4 and modulation of their fibrinolytic potential. AB - Our previous studies showed that thymosin beta4 (Tbeta4) induced the synthesis of plasminogen activator inhibitor-1 (PAI-1) in cultured human umbilical vein endothelial cells (HUVECs) via the AP-1 dependent mechanism and its enhanced secretion. In this work we provide evidence that the released PAI-1 is accumulated on the surface of HUVECs, exclusively in its active form, in a complex with alpha1-acid glycoprotein (AGP) that is also up-regulated and released from the cells. This mechanism is supported by several lines of experiments, in which expression of both proteins was analyzed by flow cytometry and their colocalization supported by confocal microscopy. PAI-1 did not bind to quiescent cells but only to the Tbeta4-activated endothelial cells. In contrast, significant amounts of AGP were found to be associated with the cells overexpressing enhanced green fluorescent protein (EGFP)-alpha1-acid glycoprotein (AGP) without Tbeta4 treatment. The AGP.PAI-1 complex was accumulated essentially at the basal surface of endothelial cells, and such cells showed (a) morphology characteristic for strongly adhered and spread cells and (b) significantly reduced plasmin formation. Taken together, these results provide the evidence supporting a novel mechanism by which active PAI-1 can be bound to the Tbeta4 activated endothelial cells, thus influencing their adhesive properties as well as their ability to generate plasmin. PMID- 16272159 TI - Insulin enhances growth hormone induction of the MEK/ERK signaling pathway. AB - Growth hormone (GH) plays an important role in growth and metabolism by signaling via at least three major pathways, including STATs, ERK1/2, and phosphatidylinositol 3-kinase/Akt. Physiological concentrations of insulin promote growth probably by modulating liver GH receptor (GHR) levels in vivo, but the possible effects of insulin on GH-induced post-GHR signaling have yet to be studied. We hypothesized that short-term insulin, similar to the fluctuations that occur following feeding, affects GH-induced post-GHR signaling. Our present studies suggest that, in rat H4IIE hepatoma cells, insulin (4 h or less) selectively enhanced GH-induced phosphorylation of MEK1/2 and ERK1/2, but not GH induced activation of STAT5 and Akt. Although insulin pretreatment altered GH induced formation of Shc.Grb2.SOS complex, it did not significantly affect GH induced activation of other signaling intermediates upstream of MEK/ERK, including JAK2, Ras, and Raf-1. Immunofluorescent staining indicated that insulin pretreatment facilitated GH-induced cell membrane translocation of MEK1/2. Insulin pretreatment also increased the amount of MEK association with its scaffolding protein, KSR. In summary, short-term insulin treatment of cultured, liver-derived cells selectively sensitized GH-induced MEK/ERK phosphorylation independent of JAK2, Ras, and Raf-1, but likely resulted from increased cell membrane translocation of MEK1/2. These findings suggest that insulin may be necessary for sensitization of cells to GH-induced ERK1/2 activation and provides a potential cellular mechanism by which insulin promotes growth. PMID- 16272161 TI - Questioning sustainability in health promotion projects and programs. PMID- 16272160 TI - Mechanism of uptake of C105Y, a novel cell-penetrating peptide. AB - C105Y, a synthetic peptide (CSIPPEVKFNKPFVYLI) based on the amino acid sequence corresponding to residues 359-374 of alpha1-antitrypsin, enhances gene expression from DNA nanoparticles. To investigate how this enhancement occurs, C105Y was fluorescently labeled to study its uptake and intracellular trafficking. When human hepatoma cells (HuH7) were incubated with fluorescently labeled C105Y for as little as 3 min, C105Y displayed nuclear and cytoplasmic staining with enrichment of fluorescent signal in the nucleus and nucleolus. Uptake and nucleolar localization were observed with the short sequence PFVYLI, but not with SIPPEVKFNK, and the D-isomer was readily taken up into cells but not into the nucleus. We found that the C105Y peptide is routed to the nucleolus very rapidly in an energy-dependent fashion, whereas membrane translocation and nuclear localization are energy-independent. When we tested the involvement of known endocytosis pathways in uptake and trafficking of this peptide, we demonstrated that C105Y peptide is internalized by a clathrin- and caveolin-independent pathway, although lipid raft-mediated endocytosis may play a role in peptide intracellular trafficking. Efficient energy-independent cell entry with rapid nuclear localization probably accounts for enhancement of gene expression from inclusion of C105Y into DNA nanoparticles. PMID- 16272162 TI - Long-term evaluation of cardiac function in children who received anthracyclines during pregnancy. AB - BACKGROUND: The use of anthracyclines in patients with cancer has been associated with the presence, even when standard doses were employed, of cardiac toxicity, most frequently after 5 years of therapy. Treatment of cancer during pregnancy remains a dilemma because cytotoxic therapy has been associated with the presence of severe side-effects. The outcome of children that received antracyclines during pregnancy, including during the first trimester, remain unknown because long-term follow-up is not available. PATIENTS AND METHODS: Eighty-one children whose mothers (29 acute leukemia, 33 malignant lymphoma and 19 Hodgkin's disease) were treated with cytotoxic drugs, including anthracyclines, during pregnancy were evaluated to detect cardiac toxicity, including clinical evaluation and echocardiogram [all parameters were evaluated, but fraction shortening (FS) was taking as the best parameter to evaluate cardiac toxicity in children] every 5 years after birth until 29 years of age. RESULTS: Children with actual age of 9.3 29.5 years (mean 17.1) did not show any clinical date of cardiac disfunction, in all cases echocardiogram was normal and FS did not showed any abnormality during the follow-up. CONCLUSIONS: The use of anthracyclines did not show any clinical or echocardiogram evidence of late cardiac toxicity. We hope that the present report increases the number of reports of the long-term follow-up of children who received cytotoxic drugs, in order to define the best treatment in this special patient setting. PMID- 16272163 TI - Voxel-wise analysis of diffusion weighted imaging reveals disruption of the olfactory tract in Parkinson's disease. AB - Diffusion weighted imaging (DWI) and the trace of diffusion tensor [Trace (D)], a marker of water molecule diffusivity, provide information on structural integrity of nervous tissues. To investigate structural lesions within the brain's neuronal architecture in early stages of Parkinson's disease, 12 patients with disease duration of 3.5 +/- 1.5 years were studied with DWI. Data were compared with 12 age-matched healthy control subjects. To objectively localize focal changes of structural neuronal integrity without having to make an a priori hypothesis as to its location statistical parametric mapping (SPM) was applied to our DWI study. SPM localized significant increases of diffusivity in the region of both olfactory tracts in patients (P < 0.001). Trace (D) cut-off values for the voxel cluster of the olfactory tracts have been calculated from the subjects entered into SPM and applied to a total of 17 different individuals (9 patients with Parkinson's disease, disease duration 3.1 +/- 1.3 years and 8 age-matched healthy subjects). Out of 17 subjects, 16 subjects, i.e. 94%, were correctly discriminated with a sensitivity of 100% and a specificity of 88%. All patients with Parkinson's disease were correctly classified and only one normal subject was classified as having the disease, underlining the high potential of this method to separate patients with the illness from healthy subjects. Increased diffusivity in the olfactory tract is in line with the well-established clinical finding of hyposmia in Parkinson's disease. Whether DWI can be used as a marker to identify individuals at risk to develop this disease remains to be shown. PMID- 16272164 TI - Mutations in the gene LRRK2 encoding dardarin (PARK8) cause familial Parkinson's disease: clinical, pathological, olfactory and functional imaging and genetic data. AB - We have established that the frequency of LRRK2 mutations in a series of 118 cases of familial Parkinson's disease is 5.1%. In the largest family with autosomal dominant, late-onset Parkinson's disease where affected subjects share a Y1699C missense mutation we provide a detailed clinical, pathological and imaging report. The phenotype in this large British kindred included asymmetrical, levodopa-responsive parkinsonism where unilateral leg tremor at onset and foot dystonia were prominent features. There was no significant abnormality of cognition but there was prominent behavioural disorder. We observed a lower age of onset in successive generations. Histopathology in one patient showed substantia nigra cell loss and Lewy body formation, with small numbers of cortical Lewy bodies. 18F-dopa positron emission tomography (PET) in another patient showed a pattern of nigrostriatal dysfunction typical of idiopathic Parkinson's disease. 18F-dopa-PET scans in unaffected family members prior to identifying the disease locus did not detect subclinical nigrostriatal dysfunction. Olfaction was assessed in affected subjects and Lewy bodies were identified in the olfactory bulb as well as cortex and brainstem of one deceased patient. In order to assess the role of mutations in this gene in other familial cases we undertook a mutation screen of all 51 exons of LRRK2 in 117 other smaller British kindreds with familial Parkinson's disease. The commonest mutation was G2019S and we also identified two novel mutations, R1941H and T2356I, in the coding sequence. These data suggest that parkinsonism caused by mutations in LRRK2 is likely to represent the commonest locus for autosomal dominant Parkinson's disease with a phenotype, pathology and in vivo imaging similar to idiopathic, late-onset Parkinson's disease. PMID- 16272165 TI - Leucodysplasia, microcephaly, cerebral malformation (LMC): a novel recessive disorder linked to 2p16. AB - We report three related and one unrelated child with an apparently novel neurodevelopmental disorder. The clinical course was very similar in all the four patients: congenital microcephaly with severe failure of post-natal brain growth, neonatal onset of intractable seizures associated with lack of developmental progression and death within the first 3 years of life. The appearance on cerebral neuroimaging was almost identical, with simplified gyration associated with a non-thickened cortex, severe hypoplasia of the corpus callosum, a small flattened brain stem, and specific cystic lesions in the white matter around the temporal and occipital horns. To our knowledge these patients represent a previously unreported, autosomal recessive syndrome. Homozygosity mapping in the consanguineous family has identified a candidate region on the chromosome 2p16. PMID- 16272166 TI - Negative myoclonus induced by cortical electrical stimulation in epileptic patients. AB - Negative myoclonus (NM) is a motor disorder characterized by a sudden and abrupt interruption of muscular activity. The EMG correlate of NM is a brief (<500 ms) silent period (SP) not preceded by any enhancement of EMG activity (i.e. myoclonus). This study investigated the role of premotor cortex (PMC), primary motor cortex (MI), primary somatosensory area (SI) and supplementary motor area (SMA) in the pathophysiology of cortical NM by means of intracerebral low frequency (1 Hz) electrical stimulation. In three drug-resistant epileptic patients undergoing presurgical evaluation, we delivered single electric pulses (stimulus duration: 3 ms; stimulus intensity ranging from 0.4 to 3 mA) to PMC (2 patients), MI (1 patient), SI and SMA through stereo-EEG electrodes; surface EMG was collected from both deltoids. The results showed that (i) the stimulation of PMC or MI could evoke a motor evoked potential (MEP) either at rest or during contraction, in this latter case followed by an SP; however, in two patients, at the lowest stimulus intensities (0.4 mA), 50% of stimuli could induce a pure SP, i.e. not preceded by an MEP; raising the intensity of stimulation (0.6 mA), the SPs showed an antecedent MEP in >80% of stimuli; (ii) the stimulation of SI at low stimulus intensities (from 0.4 to 0.8 mA) induced in two patients only SPs, never associated with an antecedent MEP, whereas in the third subject the SPs could be inconstantly preceded by an MEP; by incrementing the stimulus intensity (up to 3 mA), in all three patients the SPs tended to be preceded, although not constantly, by an MEP; stimulus intensity affected SP duration (i.e. the higher the intensity, the longer the SP), without influencing the latency of onset of the SPs; (iii) the stimulation of SMA induced only pure SPs, at all stimulus intensities up to 3 mA; as for SI, increment of stimulus intensity was paralleled by an increase in SP duration, without influencing the onset latency of SPs. We conclude that single electric pulse stimulation of PMC, MI, SI and SMA through stereo-EEG electrodes can induce pure SPs, not preceded by an MEP, which clinically appear as NM, suggesting therefore that these cortical areas may be involved in the genesis of this motor phenomenon. However, it must be pointed out that SMA stimulation induced only pure SPs, regardless of the stimulus intensity, whereas occurrence of pure SPs following stimulation of PMC, MI, and SI depended mainly on the intensity of stimulation. PMID- 16272167 TI - Apoptosis dominant in the periinfarct area of human ischaemic stroke--a possible target of antiapoptotic treatments. AB - Animal experiments have suggested that apoptotic programmed cell death is responsible for an important portion of the delayed ischaemic brain damage. Antiapoptotic signalling through erythropoietin (EPO) binding to its receptor (EPOR) is triggered by systemic or local hypoxia and may exist in the post ischaemic brain, and a neuroprotective effect by EPO was described recently and proposed for clinical stroke treatment. The objective of the study was to determine whether apoptosis occurs in human ischaemic stroke and to describe its topographical distribution. An autopsy cohort consisting of 13 cases of fatal ischaemic stroke (symptom duration from 15 h to 18 days) treated at the Department of Neurology, Helsinki University Central Hospital and 3 controls were studied. DNA damage was investigated by immunofluorescent TUNEL-labelling in combination with apoptotic cell morphology and by visualization of a major signalling system of apoptosis, Fas-FasL (Fas-ligand), by the immunoperoxidase technique. The relationship of EPO and EPOR in the face of TUNEL-labelled and necrotic cell death was co-registered in human cerebral neurons undergoing different stages of ischaemic change. TUNEL-labelled cells with apoptotic morphology were disproportionately more frequent, 148% (30) [mean (SE)] in the periinfarct versus 97% (22) in the core, as percentage of the cells in the contralateral hemisphere (P = 0.027). The apoptotic cell percentage reached up to 26% (2) of all cells in periinfarct area. A linear correlation was found for Fas and its counterpart FasL expression (r(S) = 0.774, P < 0.001). Ischaemia induced widespread neuronal expression of EPOR, which was inversely related to the severity of ischaemic neuronal necrosis (P < 0.05). To conclude, these data verify the predominance of apoptosis in the periphery of human ischaemic infarctions. Fas and FasL were linearly overexpressed supporting that this 'death receptor' complex may promote the completion of cell death. Increased EPO signalling may be a cellular response for survival in less severely damaged areas. These results support antiapoptotic therapies against delayed neuronal cell death in human ischaemic stroke. PMID- 16272168 TI - Novel retinoblastoma binding protein RBBP9 modulates sex-specific radiation responses in vivo. AB - Retinoblastoma (RB) tumor suppressor is a key regulator of apoptosis, a central mediator of the proliferative block induced by ionizing radiation (IR) and a binding target for a variety of proteins that regulate its activity. One of the recently discovered and the least investigated of these is the novel Rb-binding protein RBBP9/BOG. We studied the effects of acute and chronic low dose radiation (LDR) exposure on the induction of RBBP9 and RB signaling pathway in vivo in mouse spleen and found that RBBP9 played a pivotal role in IR responses in vivo. We observed that chronic LDR exposure led to a significant increase of RBBP9 expression in males and a significant decrease in females. Elevated RBBP9 expression in males was paralleled by a pronounced dephosphorylation of RB and a significant drop of PCNA and cyclin A expression. On the contrary, chronic exposure in females led to decreased levels of RBBP9 and increased levels of hyperphosphorylated RB (ppRB) in spleen. Decreased levels of ppRB in spleen of chronically exposed males were correlated with strongly elevated apoptotic rates. In females, the radiation-induced increase of apoptotic index was much less pronounced. Quite surprisingly, the observed sex-specific signaling changes did not result in the sex-specificity of cellular proliferation. The molecular mechanisms and possible repercussions of the radiation-induced sex differences in cellular proliferation and apoptosis are discussed. PMID- 16272169 TI - Liquid chromatography-electrospray ionization-mass spectrometry: the future of DNA adduct detection. AB - Over the past 40 years considerable emphasis has been placed on the development of accurate and sensitive methods for the detection and quantitation of DNA adducts. The formation of DNA adducts resulting from the covalent interaction of genotoxic carcinogens with DNA, derived from exogenous and endogenous sources, either directly or following metabolic activation, can if not repaired lead to mutations in critical genes such as those involved in the regulation of cellular growth and subsequent development of cancer. The major analytical challenge has been to detect levels of DNA adducts at the level of 0.1-1 adducts per 10(8) unmodified DNA bases using only low microgram amounts of DNA, and with high specificity and accuracy, in humans exposed to genotoxic carcinogens derived from occupational, environmental, dietary and life-style sources. In this review we will highlight the merits as well as discuss the progress made by liquid chromatography coupled to electrospray ionization mass spectrometry as a method for DNA adduct detection. PMID- 16272170 TI - Reduced expression of EphB2 that parallels invasion and metastasis in colorectal tumours. AB - EphB2, a receptor tyrosine kinase regulated by the beta-catenin/Tcf4 complex, is expressed in the proliferative compartment of mouse intestine and regulates bidirectional migration of intestinal precursor cells in the crypt-villus axis through repulsive interaction with Ephrin-B ligands. Recently, it has been shown that reduction of EphB activity accelerates colon tumour progression in the Apc(Min/+) mice. In this study, we examined the expression of EphB2 in normal colon, adenomas, primary colorectal cancers (CRCs), lymph node metastases and liver metastases using immunohistochemistry on tissue microarrays. In addition, EphB2 was overexpressed in SW480 colon cancer cells to study its effect in vitro. We found that EphB2 was expressed in 100% of normal colon crypt base cells, 78% of adenomas, 55.4% of primary CRCs, 37.8% of lymph node metastases and 32.9% of liver metastases (all differences were statistically significant at P < 0.001 compared with primary CRCs). Patients with CRCs that lose EphB2 expression had more advanced tumour stage (P = 0.005), poor differentiation (P < 0.001), poor overall survival (P = 0.005) and disease-free survival (P = 0.001), with the latter being independent of tumour stage. In vitro studies showed that overexpression of EphB2 inhibited colon cancer cell growth in colony formation assay and activation of EphB2 receptor inhibited colon cancer cell adhesion and migration. Our data demonstrated a progressive loss of EphB2 expression in each critical step of colon carcinogenesis, including the onset of invasion, dedifferentiation and metastasis which are paralleled by adverse patient outcome. EphB2 may achieve its tumour suppressor function through regulation of cell survival, adhesion and migration. PMID- 16272171 TI - Gene-environment interaction: the role of SULT1A1 and CYP3A5 polymorphisms as risk modifiers for squamous cell carcinoma of the oesophagus. AB - An imbalance in the activities of enzymes involved in the metabolism, conjugation and transport of xenobiotics may account for the variability in susceptibility to the development of complex diseases such as cancer between different population groups. In this study we investigated a functional polymorphism in the SULT1A1 gene in 245 patients and 288 controls. Previous studies have shown that the 638G- >A polymorphism that results in the substitution of arginine by histidine at codon 213 (SULT1A1*2) results in decreased SULT1A1 activity. The same group of samples used in this study had been previously genotyped for CYP3A5 genetic polymorphisms. Among Black subjects the burning of wood or charcoal for cooking and keeping warm was significantly associated with increased risk for oesophageal cancer (OC) (AOR, 15.2; P=0.001) as was the consumption of home-brewed beer (AOR, 6.97; P=0.0001). Among the Mixed Ancestry group, tobacco smoking combined with alcohol consumption were significantly associated with higher risk for OC (AOR, 5.18; P=0.0005). In both Blacks and Mixed Ancestry subjects, starting to smoke below the age of 20 years was associated with significantly increased risk for OC (AOR, 3.5 among the Blacks and AOR, 12 among the Mixed Ancestry). The homozygous SULT1A1*2/*2 genotype was associated with increased risk for OC among smokers. The SULT1A1*2/*2 genotype in combination with the CYP3A5 heterozygous genotypes was associated with significantly increased risk for OC (AOR, 3.60; P=0.001) with the risk being even higher among smokers compared with non-smokers. The above findings confirm the association between alcohol consumption and tobacco smoking with increased risk for OC. The genotype results show that SULT1A1*2/*2 genotype is associated with increased risk for OC among subjects exposed to tobacco-smoke related carcinogens. PMID- 16272172 TI - ERK and JNK signaling pathways are involved in the regulation of activator protein 1 and cell death elicited by three isothiocyanates in human prostate cancer PC-3 cells. AB - Many isothiocyanates (ITCs) such as sulforaphane (SFN), phenethyl isothiocyanate (PEITC) and allyl isothiocyanate (AITC) are highly effectively in chemoprevention or reduction of the risk of cancer and possess antitumor activities in vitro and in vivo. The activator protein 1 (AP-1) and MAPK signaling pathways are believed to play an important role in cancer chemoprevention and chemotherapy due to their involvement in tumor cell growth, proliferation, apoptosis and survival. In the present study, we determined the effects of SFN, PEITC and AITC on AP-1 activation, and investigated the roles of extracellular signal-regulated protein kinase (ERK) and c-Jun N-terminal kinase (JNK) signaling pathways in the regulation of AP-1 activation and cell death elicited by these ITCs in human prostate cancer PC-3 cells. SFN, PEITC and AITC each induced AP-1 activity potently and caused a significant elevation in the phosphorylation of ERK1/2, JNK1/2, Elk-1 and c-Jun. Transfection with ERK2 and upstream kinase DNEE-MEK1 activated AP-1 activity, and transfection with dominant-negative mutant ERK2 (dnERK2) potently decreased AP-1 activation induced by SFN, PEITC and AITC. Transfection with JNK1 and upstream kinase MKK7 activated AP-1 activity, and transfection with dominant-negative mutant JNK1-APF significantly attenuated AP-1 activation induced by SFN, PEITC and AITC. Pretreatment with MEK1-ERK inhibitor U0126 and JNK inhibitor SP600125 substantially attenuated the decrease in cell viability induced by SFN, PEITC and AITC. Transfection with dnERK2 and JNK1-APF significantly reversed the decrease of Bcl-2 expression elicited by these ITCs. Furthermore, transfection with dnERK2 and JNK1-APF blocked the apoptosis induced by these ITCs in PC-3 cells. Taken together, our results indicate that the activation of the ERK and JNK signaling pathways is important for transcriptional activity of AP-1 and is involved in the regulation of cell death elicited by ITCs in PC-3 cells. PMID- 16272173 TI - Genome-wide differences between microsatellite stable and unstable colorectal tumors. AB - Genomic copy number changes are frequently found in cancers and they have been demonstrated to contribute to carcinogenesis; and it is widely accepted that tumors with microsatellite instability (MSI) are genetically stable and mostly diploid. In the present study we compared the copy number alterations and the gene-expression profiles of microsatellite stable (MSS) and MSI colorectal tumors. A total number of 31 fresh-frozen primary tumors (16 MSS and 15 MSI) were used. Twenty-eight samples (15 MSS and 13 MSI) were analyzed with metaphase comparative genomic hybridization (CGH), nine of which plus one additional sample (4 MSS and 6 MSI) were further analyzed by cDNA-based array-CGH. Gene expression analysis was performed with six samples [3 MSS and 3 MSI, four of these used in metaphase CGH (mCGH) analysis] to identify differentially expressed genes possibly located in the lost or amplified regions found by CGH, stressing the biological significance of copy number changes. Metaphase and array-CGH analysis of two colon cancer cell lines (HTC116 and SW480, reported as MSI and MSS archetypes) gave comparable results. Alterations found by mCGH in MSS tumors were +20, +8q, -8p and -18q. Interestingly, 1p22, 4q26 and 15q21 were found deleted preferentially in MSS tumors, while 22q13 was found gained in MSI tumors. The regions of alterations identified by array-CGH were gains at 8q24, 16q24.3 and 20q13, and the loss of 5q21, appearing in the both types of tumors. Gene expression analysis revealed genes with specific associations with the copy number changes of the corresponding genomic regions. As a conclusion, colorectal cancer is a heterogeneous disease, demonstrated by the genomic profiles of individual samples. However, our data shows that copy number changes do not occur exclusively in the MSS phenotypes. PMID- 16272174 TI - Involvement of phospholipase A2 in Pseudomonas aeruginosa-mediated PMN transepithelial migration. AB - Inflammation resulting from bacterial infection of the respiratory mucosal surface during pneumonia and cystic fibrosis contributes to pathology. A major consequence of the inflammatory response is recruitment of polymorphonuclear cells (PMNs) to the infected site. To reach the airway, PMNs must travel through several cellular and extracellular barriers, via the actions of multiple cytokines, chemokines, and adhesion molecules. Using a model of polarized lung epithelial cells (A549 or Calu-3) grown on Transwell filters and human PMNs, we have shown that Pseudomonas aeruginosa induces PMN migration across lung epithelial barriers. The process is mediated by epithelial production of the eicosanoid hepoxilin A(3) (HXA(3)) in response to P. aeruginosa infection. HXA(3) is a PMN chemoattractant metabolized from arachidonic acid (AA). Given that release of AA is believed to be the rate-limiting step in generating eicosanoids, we investigated whether P. aeruginosa infection of lung epithelial cells resulted in an increase in free AA. P. aeruginosa infection of A549 or Calu-3 monolayers resulted in a significant increase in [(3)H]AA released from prelabeled lung epithelial cells. This was partially inhibited by PLA(2) inhibitors ONO-RS-082 and ACA as well as an inhibitor of diacylglycerol lipase. Both PLA(2) inhibitors dramatically reduced P. aeruginosa-induced PMN transmigration, whereas the diacylglycerol lipase inhibitor had no effect. In addition, we observed that P. aeruginosa infection caused an increase in the phosphorylation of cytosolic PLA(2) (cPLA(2)), suggesting a mechanism whereby P. aeruginosa activates cPLA(2) generating free AA that may be converted to HXA(3), which is required for mediating PMN transmigration. PMID- 16272175 TI - Thiol oxidation inhibits nitric oxide-mediated pulmonary artery relaxation and guanylate cyclase stimulation. AB - The mechanisms through which thiol oxidation and cellular redox influence the regulation of soluble guanylate cyclase (sGC) are poorly understood. This study investigated whether promoting thiol oxidation via inhibition of NADPH generation by the pentose phosphate pathway (PPP) with 1 mM 6-aminonicotinamide (6-AN) or the thiol oxidant diamide (1 mM) alters sGC activity and cGMP-associated relaxation to nitric oxide (NO) donors [S-nitroso-N-acetylpenicillamine (SNAP) and spermine-NONOate]. Diamide and 6-AN inhibited NO-elicited relaxation of endothelium-denuded bovine pulmonary arteries (BPA) and stimulation of sGC activity in BPA homogenates. Treatment of BPA with the thiol reductant DTT (1 mM) reversed inhibition of NO-mediated relaxation and sGC stimulation by 6-AN. The increase in cGMP protein kinase-associated phosphorylation of vasodilator stimulated phosphoprotein on Ser239 elicited by 10 microM SNAP was also inhibited by diamide. Activation of sGC by SNAP was attenuated by low micromolar concentrations of GSSG in concentrated, but not dilute, homogenates of BPA, suggesting that an enzymatic process contributes to the actions of GSSG. Relaxation to agents that function through cAMP (forskolin and isoproterenol) was not altered by inhibition of the pentose phosphate pathway or diamide. Thus a thiol oxidation mechanism controlled by the regulation of thiol redox by NADPH generated via the pentose phosphate pathway appears to inhibit sGC activation and cGMP-mediated relaxation by NO in a manner consistent with its function as an important physiological redox-mediated regulator of vascular function. PMID- 16272176 TI - Hemorrhagic shock-activated neutrophils augment TLR4 signaling-induced TLR2 upregulation in alveolar macrophages: role in hemorrhage-primed lung inflammation. AB - Hemorrhagic shock renders patients susceptible to the development of acute lung injury in response to a second inflammatory stimulus by as yet unclear mechanisms. We investigated the role of neutrophils (PMN) in alveolar macrophage (AMphi) priming, specifically, the role in mediating Toll-like receptor (TLR)4 and TLR2 cross talk in AMphi. Using a mouse model of hemorrhagic shock followed by intratracheal administration of LPS, we explored a novel function of shock activated PMN in the mechanism of TLR2 upregulation induced by LPS-TLR4 signaling in AMphi. We showed that antecedent hemorrhagic shock enhanced LPS-induced TLR2 upregulation in AMphi. In neutropenic mice subjected to shock, the LPS-induced TLR2 expression was significantly reduced, and the response was restored upon repletion with PMN obtained from shock-resuscitated mice but not by PMN from sham operated mice. These findings were recapitulated in mouse AMphi cocultured with PMN. The enhanced TLR2 upregulation in AMphi augmented the expression of macrophage inflammatory protein-2, TNF-alpha, and macrophage migration inhibitory factor in the AMphi in response to sequential challenges of LPS and peptidoglycan, a prototypical TLR2 ligand, which physiologically associated with amplified AMphi-induced PMN migration into air pouch and lung alveoli. Thus TLR2 expression in AMphi, signaled by TLR4 and regulated by shock-activated PMN, is an important positive-feedback mechanism responsible for shock-primed PMN infiltration into the lung after primary PMN sequestration. PMID- 16272178 TI - Experimental medicine in psychiatry. PMID- 16272177 TI - Adherence of airway neutrophils and inflammatory response are increased in CF airway epithelial cell-neutrophil interactions. AB - Persistent presence of PMN in airways is the hallmark of CF. Our aim was to assess PMN adherence, percentage of apoptotic airway PMN (aPMN), and IL-6 and IL 8 production when aPMN are in contact with airway epithelial cells. Before coculture, freshly isolated CF aPMN have greater spontaneous and TNF-alpha induced apoptosis compared with blood PMN from the same CF patients and from aPMN of non-CF patients. We then examined cocultures of PMN isolated from CF and non CF airways with bronchial epithelial cells bearing mutated cftr compared with cftr-corrected bronchial epithelial cells. After 18-h coculture, the number of CF aPMN adhered on cftr-deficient bronchial epithelial cells was 2.3-fold higher compared with the coculture of non-CF aPMN adhered on cftr-corrected bronchial epithelial cells. The percentage of CF apoptotic aPMN (9.5 +/- 0.2%) adhered on cftr-deficient bronchial epithelial cells was similar to the percentage of non-CF apoptotic aPMN adhered on cftr-corrected bronchial epithelial cells (10.3 +/- 0.7%). IL-6 and IL-8 levels were enhanced 6.5- and 2.9-fold, respectively, in coculture of CF aPMN adhered on cftr-deficient bronchial epithelial cells compared with coculture of non-CF aPMN adhered on cftr-corrected bronchial epithelial cells. Moreover, blocking surface adhesion molecules ICAM-1, VCAM-1, and E-selectin on cftr-deficient bronchial epithelial cells with specific MAbs inhibited the adherence of CF aPMN by 64, 51, and 50%, respectively. Our data suggest that in CF patients a high number of nonapoptotic PMN adhered on airway epithelium associated with elevated IL-6 and IL-8 levels may contribute to sustained and exaggerated inflammatory response in CF airways. PMID- 16272179 TI - Evidence-based guidelines for the pharmacological treatment of anxiety disorders: recommendations from the British Association for Psychopharmacology. AB - These British Association for Psychopharmacology guidelines cover the range and aims of treatment for anxiety disorders. They are based explicitly on the available evidence and are presented as recommendations to aid clinical decision making in primary and secondary medical care. They may also serve as a source of information for patients and their carers. The recommendations are presented together with a more detailed review of the available evidence. A consensus meeting involving experts in anxiety disorders reviewed the main subject areas and considered the strength of evidence and its clinical implications. The guidelines were constructed after extensive feedback from participants and interested parties. The strength of supporting evidence for recommendations was rated. The guidelines cover the diagnosis of anxiety disorders and key steps in clinical management, including acute treatment, relapse prevention and approaches for patients who do not respond to first-line treatments. PMID- 16272180 TI - Actions of antipsychotic drugs on pancreatic beta-cell function: contrasting effects of clozapine and haloperidol. AB - The use of antipsychotic drugs is known to be associated with a number of adverse metabolic side effects, including diabetes mellitus. These side effects could be, at least in part, the result of impaired islet cell function, although the underlying mechanisms are unknown. We have studied the effects of the atypical antipsychotic clozapine and of the conventional drug haloperidol on electrical and secretory activity in rat pancreatic beta-cells. At a low glucose concentration (4mM), clozapine (5microM) had little or no effect on membrane potential. However, at a stimulatory glucose concentration (16mM), clozapine was found to hyperpolarize the cell membrane potential, resulting in a complete inhibition of electrical activity. In contrast, haloperidol (5microM) was found to cause a marked depolarization of the membrane potential in the presence of both low and high concentrations of glucose. Clozapine and haloperidol were found, respectively, to increase and decrease-cell input conductance, an index of K(+) permeability. Single channel recordings indicated that changes in K(ATP) channel activity contributed towards these effects. Neither clozapine nor haloperidol affected basal insulin release, although clozapine inhibited glucose induced insulin release. It is concluded that clozapine and haloperidol exert contrasting actions on electrical activity in rat pancreatic -cells as a result of opposing effects on K(+) permeability. These findings may relate to the increased incidence of diabetes associated with clozapine treatment. PMID- 16272181 TI - Does the human leukaemia differentiation factor fragment HLDF6 improve memory via brain DNA and protein synthesis? AB - The novel human differentiating factor peptide fragment HLDF6 (Thr-Gly-Glu-Asn His-Arg) was synthesized and purified. HLDF6 (0.1mg/kg i.p. but not 1mg/kg i.p.) improved not only long-term (24h) memory in adult rats in the water maze behavioural paradigm but also performance in the delayed matching-to-position (DMTP) task (0.3 and 1.0 but not 0.1mg/kg i.p). Hence, HLDF6 not only enhanced allocentric spatial learning and reference memory (water maze) but also improved temporal, spatial and working memory processes in the DMTP behavioural paradigm. Immunoreactivity blotting analysis of HLDF (the protein precursor of HLDF6) was performed and the following rank order of visual intensities from brain structures was noted: hippocampus cerebral cortex cerebellum hypothalamus striatum. Subsequently, we found that the highest absolute levels of HLDF were expressed in the hippocampus and cerebral cortex as detected by ELISA. We also demonstrated that HLDF6 enhanced [(3)H]-thymidine and [(14)C]-leucine incorporation into whole brain and hippocampal homogenates (maxima occurring within the range 10 (-12)-10 (-6) M) suggesting that this hexapeptide promoted de novo DNA and protein biosynthesis. We discuss this data in terms of their implications for links with other integrative metabolic pathways involving immediate early gene activation which may underpin a potential application for HLDF6 in limiting memory impairments associated with neurodegenerative diseases. PMID- 16272182 TI - The use of sleep measures to compare a new 5HT1A agonist with buspirone in humans. AB - The partial agonist buspirone has a REM (rapid eye movement) suppressing effect on human sleep probably via a 5HT(1A) receptor in the pontine area. Eptapirone is a new 5HT(1A) agonist with a greater intrinsic effect than buspirone. The objective of this study was to examine the effects of eptapirone on sleep architecture, particularly REM sleep, in normal volunteers and compare it with buspirone and placebo. This was a randomized, double-blind placebo-controlled four-way crossover study in 12 healthy volunteers. Volunteers were screened to ensure that they had normal overnight sleep EEG (electroencephalogram) and were extensive CYP 2D6 metabolizers. Sleep was recorded on pairs of nights on four occasions, with medication being taken before the second night. Treatments were eptapirone 1.5mg at 10 AM, eptapirone 1.5mg at 11 PM, buspirone 20mg at 11 PM and placebo. Standard measures of sleep were derived and compared among the four treatments using ANOVA. REM sleep was significantly suppressed supporting the proposition that activation of post-synaptic 5HT(1A) receptors reduces REM sleep. Sleep fragmentation increased by both drugs. REM sleep suppression was significantly greater with morning eptapirone than with buspirone. Wakefulness in sleep was significantly greatest after morning eptapirone. REM sleep effects were greatest after evening eptapirone, suggesting a greater effect on central serotonin receptors than that of buspirone. PMID- 16272184 TI - Effects of repeated doses of caffeine on mood and performance of alert and fatigued volunteers. AB - Evidence for behavioural effects of caffeine is well documented in the literature. It is associated with increased subjective alertness, improved reaction time and enhanced encoding of new information. These effects are most prominent in low arousal situations. However, there is an ongoing debate as to whether such changes are in fact improvements or merely a reversal of the negative effects of a period of caffeine withdrawal (e.g. overnight abstinence). To avoid such a confound this study included multiple doses of caffeine which were administered under double-blind conditions to participants who had ingested their normal daily quota of caffeine. In the present study participants were fatigued by carrying out a prolonged testing schedule in the evening. Sixty volunteers, all regular caffeine consumers, took part in the study. They attended for three sessions on separate days. They were instructed to consume normal amounts of caffeinated beverages. Consumption was measured by a diary and saliva samples were taken and caffeine assays conducted. A baseline test session was carried out at 18.00h and following this a double blind placebo controlled caffeine challenge (1.5mg/kg) conducted. The test battery was repeated twice approximately 30 minutes after the caffeine challenge. Following this another drink was administered and the test battery repeated twice more. On one test session volunteers had placebo in both drinks, in another they had caffeine in both drinks and another caffeine in the first and placebo in the second. Order of conditions was balanced across subjects. The results showed that caffeine led to a more positive mood and improved performance on a number of tasks. Different effects of caffeine were seen depending on the person's level of arousal. Linear effects of caffeine dose were also observed. This is evidence against the argument that behavioural changes due to caffeine are merely the reversal of negative effects of a long period of caffeine abstinence. The findings are discussed in relation to both noradrenergic and cholinergic neurotransmitter systems. PMID- 16272183 TI - Gender differences in response to lorazepam in a human drug discrimination study. AB - Gender differences in the discriminative stimulus properties of drugs of abuse have sometimes been reported, although we have previously found no differences in subjective or discriminative responses in human subjects acquiring an alcohol discrimination. The aim of the present work was to determine if there were gender differences in the effects of lorazepam, a benzodiazepine-receptor agonist which substituted for the alcohol stimulus in trained social drinkers. Volunteers who had already acquired an alcohol (0.2g/kg) placebo discrimination were administered (double-blind) either placebo (nine females, nine males) or lorazepam 2mg (six females, six males). They then sampled a series of five drinks and rated each one for likeness to the training stimulus (the generalization response). In addition they completed rating scales for subjective effects and the Digit Symbol Substitution Test (DSST). Lorazepam substituted for the alcohol stimulus equally in both sexes and increased associated scores for lightheadedness. Females however, showed a much greater DSST performance impairment following lorazepam, compared with males. This effect was independent of body weight differences and sedation. These results are discussed in the light of current knowledge of gender differences in response to drugs of abuse and suggest that the stimulus and cognitive effects of benzodiazepine-receptor agonists are modulated by different brain mechanisms. PMID- 16272185 TI - Effects of ethanol on kinaesthetic perception. AB - In normal subjects, alcohol increases handwriting size, but the mechanism is not understood. Here we show that the alcohol effect on handwriting can be explained by a selective impairment of kinaesthetic perception. Thirty volunteers (15 male, aged 18-29 years) took part in an open study. They were tested before and after a drink containing vodka intended to produce a blood alcohol concentration of about 80mg/100ml. Tests included kinaesthetic distance estimation, in which volunteers worked with preferred hand and arm behind a screen which hid their movements; visual distance estimation; and measures of handwriting and drawing. Blood alcohol concentration at 55min, based on breathalyser measurements, was 76.7mg/100ml (SD 9.8). When asked to move the hand and mark a distance of 10cm from a starting point, distances estimates increased by 7-10% (p 0.01). Similar increases were seen for writing words and drawing characters. Signatures were increased in height but not in length. Distances estimated visually were increased much less, by 3-4% (p 0.05). Tests of psychomotor performance indicated the expected effects of ethanol. These results suggest that ethanol affects writing size by reducing kinaesthetically perceived distances. PMID- 16272186 TI - Transient memory impairment after acute dose of 75mg 3.4-Methylene dioxymethamphetamine. AB - A range of studies has indicated that users of 3.4-Methylene-dioxymethamphetamine (MDMA, 'Ecstasy') display cognitive deficits, particularly memory impairment, as compared to non-drug using controls. Yet it is difficult to determine whether these deficits are caused by MDMA or some other confounding factor, such as polydrug use. The present study was designed to establish the direct relation between MDMA and memory impairment under placebo-controlled conditions. Eighteen recreational MDMA users participated in a double blind, placebo controlled, 3-way crossover design. They were treated with placebo, MDMA 75mg and methylphenidate 20mg. Memory tests were conducted between 1.5-2h (intoxication phase) and between 25.5-26h (withdrawal phase) post dosing. Results showed that a single dose of MDMA caused impairment of immediate and delayed recall on a verbal learning task during the intoxication phase. However, there was no residual memory impairment during the withdrawal phase. Subjects reported more fatigue and less vigour, but no symptoms of depression during the withdrawal phase of MDMA treatment. Methylphenidate did not affect memory or mood at any time of testing. A single dose of MDMA produces transient memory impairment. PMID- 16272187 TI - Individual differences in aggressive responding to intravenous flumazenil administration in adult male parolees. AB - Nonhuman and human studies have shown that benzodiazepine (BZD) receptor agonists can modify aggressive behaviour. However, it is unknown whether flumazenil, a BZD receptor antagonist, enhances or inhibits aggressive behaviour. The present study was designed to investigate the effects of acute administrations of flumazenil on aggressive responding in adult humans. Six adult males with histories of childhood conduct disorder (DSM IV R) participated in experimental sessions. Aggression was measured using the Point Subtraction Aggression Paradigm (PSAP; Cherek 1992), which provided subjects with aggressive and monetary-reinforced response options. Acute doses of flumazenil (2 and 3mg) did not produce statistically significant changes in either monetary-reinforced responding or aggressive responding. The analysis of individual subjects data revealed that aggressive responses varied across subjects. The results are discussed in terms of individual differences based on the previous history of BZD abuse. Additional laboratory research is needed to better clarify the behavioural mechanisms by which BZD receptor antagonists modify human aggressive responding. PMID- 16272188 TI - Chronic treatment with modafinil may not be beneficial in patients with chronic fatigue syndrome. AB - Fourteen patients (7 male, 7 female, 22-63 years), classified as having chronic fatigue syndrome (CFS), but without concurrent major depression, significant sleepiness or use of psychoactive medication, completed a double-blind, placebo controlled, crossover study of the effects of the selective wakefulness-promoting agent, modafinil (200 and 400mg/day). The treatment periods were each 20 days, with washout periods of 2 weeks. The primary aim was to determine effects on cognition and the secondary aim was to determine effects on self-ratings of fatigue, quality of life and mood. Modafinil had mixed effects in two cognitive tasks. In a test of sustained attention, treatment with 200mg reduced the latency to correctly detect sequences, but 400mg increased the number of missed targets. In a test of spatial planning, the 200mg dose resulted in a slower initial thinking time for the easiest part of the task, whereas 400mg reduced the initial thinking time for the hardest part of the test. Lastly, in a test of mental flexibility and one of motor speed, patients performed worse whilst on modafinil (400mg), compared with the placebo period. No effects were observed on the performance of other psychometric tests or on self-ratings of fatigue, quality of life or mood, but this may have been due to insufficient statistical power. It is discussed whether the limited and mixed cognitive effects that we observed could have occurred by chance, or whether a subgroup of CFS patients with daytime sleepiness would have shown greater benefits. PMID- 16272189 TI - The efficacy and safety of quetiapine for treatment of geriatric psychosis. AB - Quetiapine, an atypical antipsychotic, is effective for psychosis in younger patients, with limited adverse effects reported. This open-label naturalistic study was conducted to assess the 4-week efficacy and safety of quetiapine for treatment of geriatric psychosis. Clinical efficacy was evaluated using the Brief Psychiatric Rating Scale (BPRS) and Clinical Global Impression Improvement (CGI I) instruments before and after 4 weeks of quetiapine treatment. The sample population consisted of 100 geropsychiatric inpatients with psychosis, with the therapeutic evaluation completed by 91. Eighty-one of these 91 patients (89.0%) experienced mild-to-substantial improvement, as determined from the CGI-I. Further, a mean reduction in BPRS score of 39.5% (from baseline) was also determined. The mean daily dose of quetiapine for the fourth week was 276.1 177.2mg/day (range 50-800). Higher quetiapine dosages were administered for patients with functional psychoses compared to an analogous group with organic mental disorders. The most common adverse effects were somnolence (30.0%), lower limb weakness (28.0%) and dizziness (27.0%). Body weight and fasting triglyceride were significantly elevated after quetiapine treatment (2.2% and 8.9% from baseline, respectively). Based on the results of this study, it appears that quetiapine is an efficacious and safe treatment for geriatric inpatients with psychosis, however, there is a wide dosing range and optimal dosage is diagnosis dependent. PMID- 16272190 TI - Fatal poisoning with antipsychotic drugs, England and Wales 1993-2002. AB - Prescription of atypical antipsychotics has increased in recent years. There have also been changes in the guidance on using older drugs, particularly the restriction in the use of thioridazine. We analysed deaths due to poisoning involving antipsychotics in England and Wales, 1993-2002, by age, sex, intent, and agents involved. We also studied antipsychotic prescribing in the community and poisoning deaths in England. Deaths attributed to adverse reactions in the course of normal treatment were not studied because these deaths are not classified as 'poisonings'. The number of deaths involving antipsychotics increased from around 55 per year 1993-1998 to 74 in 2000, and then fell to 53 in 2002. Around 25% of deaths had a verdict of accidental death and in about 60% of deaths a verdict of suicide or an open verdict was recorded. There were no deaths involving thioridazine in 2002, following its removal from use in 2001. However, the number of deaths associated with atypicals, most notably olanzapine and clozapine, has increased. Age-specific death rates were highest in those aged 30 39 and 40-49 years, and were very low in those aged under 20 and 70 or over. Death rates in males were greater than in females. For many drugs the proportion mentions either alone or with ethanol, was 25-45%, but for clozapine and olanzapine such mentions totalled 65-69%. Deaths per million prescriptions (clozapine excluded) were highest for quetiapine (31.3 per million), chlorpromazine (29.4 per million) and thioridazine (15.5 per million). PMID- 16272192 TI - Paroxetine (Paxil) and congenital malformations. PMID- 16272191 TI - New trends in the cyber and street market of recreational drugs? The case of 2C-T 7 ('Blue Mystic'). AB - 2C-T-7 ('Blue Mystic'), an illicit compound which shows similarities with MDMA and other designer drugs, has been only occasionally identified in the EU, but discussion on the Internet between experimenters has recently grown significantly. We aimed at collecting together in a review the available information on 2C-T-7, both at the cyber and at the street market level. 2C-T-7 was first synthesized in 1986; its desired effects include both a sense of empathy and of well-being. Hallucinations, nausea, anxiety, panic attacks and paranoid ideation are anecdotally reported. According to the different European sources here approached, the availability of 2C-T-7 at street level seems to be currently very low, although one death related to a mono-intoxication with 2C-T-7 has been documented in the USA. With respect to information on 2C-T-7 available online, due to both redundancy and relevance issues the initial identified sample of 360 was reduced to 118 websites. In 14 (11.9%) websites, the detailed description of the 2C-T-7 synthesis was given. Harm Reduction websites appeared significantly earlier in the search engines results' list than Anti drugs (p 0.006) websites. Five (4.2%) websites apparently offered 2C-T-7 for sale. The large body of knowledge available online seems to contrast with small numbers of seizures at street level; an exhaustive web mapping of drug-related issues may be of interest for the clinician. Projects aimed at designing more 'attractive' prevention websites should be planned and future studies should better assess the characteristics of those consumers who take advantage of the online information of hallucinogenic compounds. PMID- 16272193 TI - Cost and outcome analysis of two alcohol detoxification services. AB - AIM: To examine the relationship between service use and outcomes (individual and wider consequences) using an economic analysis of a direct-access alcohol detoxification service in Manchester (the Smithfield Centre) and an NHS partial hospitalization programme in Newcastle upon Tyne (Newcastle and North Tyneside Drug and Alcohol Service, Plummer Court). METHODS: A total of 145 direct-access admissions to the Smithfield Centre and 77 admissions to Plummer Court completed a battery of questionnaires shortly after intake and were followed up 6 months after discharge. Full economic data at follow-up were available for 54 Smithfield admissions and 49 Plummer Court admissions. RESULTS: Mean total cost of treatment per patient was pound1113 at the Smithfield Centre and pound1054 at Plummer Court in 2003-04 prices. Comparing the 6 months before treatment with the 6 months before follow-up, social costs fell by pound331 on average for each patient at Plummer Court but rose by pound1047 for each patient at the Smithfield Centre. When treatment costs and wider social costs were combined, the total cost to society at Smithfield was on average pound2159 per patient whilst at Plummer Court it was pound723 per patient. Combining the cost of treatment with drinking outcomes yielded a net cost per unit reduction in alcohol consumption of pound1.79 at Smithfield and pound1.68 at Plummer Court. CONCLUSIONS: Both services delivered a flexible needs-based service to very disadvantaged population at a reasonable cost and were associated with statistically significant reductions in drinking. For some patients, there was evidence of public sector resource savings but for others these detoxification services allowed those not previously in contact with services to meet health and social care needs. These patterns of cost through time are more complex than in previous evaluations of less severely dependent patients and difficult to predict from drinking patterns or patient characteristics. More research is required to judge the suitability of generic health state measures commonly in use for health economic evaluations for assessing the short-term outcomes of alcohol treatment. PMID- 16272194 TI - Mechanisms of low-density lipoprotein-induced expression of connective tissue growth factor in human aortic endothelial cells. AB - Hyperlipidemia is a recognized risk factor for atherosclerotic vascular disease. The underlying mechanisms that link lipoproteins and vascular disease are undefined. Connective tissue growth factor (CTGF) is emerging as a key determinant of progressive fibrotic diseases, and its expression is upregulated by diabetes. To define the mechanisms through which low-density lipoproteins (LDL) promote vascular injury, we evaluated whether LDL can modulate the expression of CTGF and collagen IV in human aortic endothelial cells (HAECs). Treatment of HAECs with LDL (50 microg/ml) for 24 h produced a significant increase in the mRNA and the protein levels of CTGF and collagen IV compared with unstimulated controls. To explore the mechanisms by which LDL regulates CTGF and collagen IV expression in HAECs, we determined first if CTGF and collagen IV are downstream targets for regulation by transforming growth factor-beta (TGF-beta). The results demonstrated that TGF-beta produced a concentration-dependent increase in the protein levels of CTGF. To assess whether the induction of CTGF in response to LDL is mediated via autocrine activation of TGF-beta, HAECs were treated with LDL for 24 h in the presence and absence of anti-TGF-beta neutralizing antibodies (anti-TGF-beta NA). The results demonstrated that the increase in CTGF induced by LDL was significantly inhibited by the anti-TGF-beta NA. To investigate the upstream mediators of TGF-beta on activity of CTGF in response to LDL, HAECs were treated with LDL for 24 h in the presence and absence of cell-permeable MAPK inhibitors. Inhibition of p38(mapk) activities did not affect LDL-induced TGF-beta1, CTGF, and collagen IV expression. On the other hand, SP-600125, a specific inhibitor of c-Jun NH(2)-terminal kinase, suppressed LDL-induced TGF-beta, CTGF, and collagen IV expression, and PD-98059, a selective inhibitor of p44/42(mapk), suppressed LDL-induced TGF-beta and CTGF expression. These findings are the first to implicate the MAPK pathway and TGF-beta as key players in LDL signaling, leading to CTGF and collagen IV expression in HAECs. The data also point to a potential mechanistic pathway through which lipoproteins may promote vascular injury. PMID- 16272195 TI - Castration inhibits exercise-induced accumulation of Hsp70 in male rodent hearts. AB - Intense exercise leads to accumulation of the inducible member of the 70-kDa family of heat shock proteins, Hsp70, in male, but not female, hearts. Estrogen is at least partially responsible for this difference. Because androgen receptors are expressed in the heart and castration leads to decreases in calcium regulatory proteins and altered cardiac function, testosterone (T) or its metabolites could also be involved. We hypothesized that removal of endogenous T production through castration would reduce cardiac Hsp70 accumulation after an acute exercise bout, whereas castrated animals supplemented with 5alpha dihydrotestosterone (DHT) would show the intact male response. Fifty-four 8-wk old male Sprague-Dawley rats were divided into intact, castrated, or castrated + DHT groups (n = 18/group). At 11 wk of age, 12 animals in each group undertook a 60-min bout of treadmill running at 30 m/min (2% incline) while the remaining 6 in each group remained sedentary. At 30 min or 24 h after exercise (n = 6/time point), blood and hearts were harvested for analysis. Serum T was undetectable in castrated and DHT-treated castrated rats, whereas serum DHT was significantly reduced in castrated animals only (approximately 60% reduction) (P < 0.05). Although there were no differences in constitutive levels of Hsp70 protein, exercise significantly increased cardiac hsp70 mRNA and protein in intact and DHT supplemented rats, but not in castrated animals (P < 0.05). To examine whether castration eliminated the ability to respond to stress, another six intact and six castrated animals were subjected to a 15-min period of hyperthermia (core temperature raised to 42 degrees C) and killed 24 h later. As opposed to exercise, castrated animals subjected to heat shock exhibited increases in Hsp70 above nonshocked (i.e., sedentary) animals, similarly to intact males (P < 0.05). These data suggest that androgens, in addition to estrogen, play a role in the sexual dimorphism observed in the stress response to exercise but not heat shock. PMID- 16272196 TI - Chlamydia pneumoniae in foci of "early" calcification of the tunica media in arteriosclerotic arteries: an incidental presence? AB - Only a few previous works investigated the involvement of Chlamydia pneumoniae (Chlamydophila pneumoniae) in arterial calcification. The present study investigated a possible association between C. pneumoniae and medial calcification. Carotid artery segments obtained by endarterectomy from 60 patients were examined by PCR and immunohistochemistry to identify the presence of C. pneumoniae. Arterial specimens showing double-positive (n = 17), double negative (n = 22), and single-positive results (n = 21) were further analyzed by a combination of histology, immunohistochemistry, and electron microscopy. Medial calcification occurred in 10 of 17 (58.8%) C. pneumoniae double-positive arterial specimens, but no medial calcification was observed in any of 22 C. pneumoniae double-negative arterial specimens. Electron microscopy indicated C. pneumoniae in smooth muscle cells (SMCs) in foci of medial calcification. Medial SMCs showing damage to the cytoplasm and basement membrane contained the structures with the appearance of elementary, reticulate, and aberrant bodies of C. pneumoniae. The presence of C. pneumoniae in SMCs was confirmed by electron microscopic immunocytochemistry. In the extracellular matrix, calcification was observed in C. pneumoniae aberrant bodies that exited the SMCs. The findings offer a new hypothesis of arterial calcification: they suggest that C. pneumoniae infection of medial SMCs may be associated with the pathophysiological events of arteriosclerotic calcification of the tunica media. PMID- 16272197 TI - Dissociation of endothelial nitric oxide synthase phosphorylation and activity in uterine artery endothelial cells. AB - Pregnancy enhanced nitric oxide production by uterine artery endothelial cells (UAEC) is the result of reprogramming of both Ca(2+) and kinase signaling pathways. Using UAEC derived from pregnant ewes (P-UAEC), as well as COS-7 cells transiently expressing ovine endothelial nitric oxide synthase (eNOS), we investigated the role of phosphorylation of five known amino acids following treatment with physiological calcium-mobilizing agent ATP and compared with the effects of PMA (also known as TPA) alone or in combination with ATP. In P-UAEC, ATP stimulated eNOS activity and phosphorylation of eNOS S617, S635, and S1179. PMA promoted eNOS phosphorylation but without activation. PMA and ATP cotreatment attenuated ATP-stimulated activity despite no increase in phospho (p)-T497 and potentiation of p-S1179. In COS-7 cells, PMA inhibition of ATP-stimulated eNOS activity was associated with p-T497 phosphorylation. Although T497D eNOS activity was reduced to 19% of wild-type eNOS with ATP and 44% with A23187, we nonetheless observed more p-S1179 with ATP than with A23187 (3.4-fold and 1.8-fold of control, respectively). Furthermore, the S1179A eNOS mutation partly attenuated ATP- but not A23187-stimulated activity, but when combined with T497D, no further reduction of eNOS activity was observed. In conclusion, although phosphorylation of eNOS is associated with activation in P-UAEC, no single or combination of phosphorylation events predict activity changes. In COS-7 cells, phosphorylation of T497 can attenuate activity but also influences S1179 phosphorylation. We conclude that in both cell types, observed changes in phosphorylation of key residues may influence eNOS activation but are not sufficient alone to describe eNOS activation. PMID- 16272198 TI - Kallikrein activates bradykinin B2 receptors in absence of kininogen. AB - Kallikreins cleave plasma kininogens to release the bioactive peptides bradykinin (BK) or kallidin (Lys-BK). These peptides then activate widely disseminated B2 receptors with consequences that may be either noxious or beneficial. We used cultured cells to show that kallikrein can bypass kinin release to activate BK B2 receptors directly. To exclude intermediate kinin release or kininogen uptake from the cultured medium, we cultured and maintained cells in medium entirely free of animal proteins. We compared the responses of stably transfected Chinese hamster ovary (CHO) cells that express human B2 receptors (CHO B2) and cells that coexpress angiotensin I-converting enzyme (ACE) as well (CHO AB). We found that BK (1 nM or more) and tissue kallikrein (1-10 nM) both significantly increased release of arachidonic acid beyond unstimulated baseline level. An enzyme-linked immunoassay for kinin established that kallikrein did not release a kinin from CHO cells. We confirmed the absence of kininogen mRNA with RT-PCR to rule out kininogen synthesis by CHO cells. We next tested an ACE inhibitor for enhanced BK receptor activation in the absence of kinin release and synthesized an ACE resistant BK analog as a control for these experiments. Enalaprilat (1 microM) potentiated kallikrein (100 nM) in CHO AB cells but was ineffective in CHO B2 cells that do not bear ACE. We concluded that kallikrein activated B2 receptors without releasing a kinin. Furthermore, inhibition of ACE enhanced the receptor activation by kallikrein, an action that may contribute to the manifold therapeutic effects of ACE inhibitors. PMID- 16272199 TI - Evidence of preserved endothelial function and vascular plasticity with age. AB - We sought to identify the relationship between shear stimuli and flow-mediated vasodilation and to determine whether small muscle mass exercise training could provoke limb-specific improvements in endothelial function in older subjects. In five young (22 +/- 1 yr old) and six old (71 +/- 2 yr old) subjects, ultrasound Doppler measurements were taken in the arm (brachial artery) and leg (deep and superficial femoral arteries) after suprasystolic cuff occlusion with and without ischemic exercise to evaluate flow-mediated dilation (FMD) in both limbs. Older subjects were reevaluated after 6 wk of single-leg knee extensor exercise training. Before the training, a significant FMD was observed in the arm of young (3 +/- 1%) but not old (1 +/- 1%) subjects, whereas a significant leg FMD was observed in both groups (5 +/- 1% old vs. 3 +/- 1% young). However, arm vasodilation was similar between young and old when normalized for shear rate, and cuff occlusion with superimposed handgrip exercise provoked additional shear, which proportionately improved the FMD response in both groups. Exercise training significantly improved arm FMD (5 +/- 1%), whereas leg FMD was unchanged. However, ischemic handgrip exercise did not provoke additional arm vasodilation after training, which may indicate an age-related limit to shear-induced vasodilation. Together, these data demonstrate that vascular reactivity is dependent on limb and degree of shear stimuli, challenging the convention of diminished endothelial function typically associated with age. Likewise, exercise training improved arm vasodilation, indicating some preservation of vascular plasticity with age. PMID- 16272200 TI - Reduced Ca2+-dependent activation of large-conductance Ca2+-activated K+ channels from arteries of Type 2 diabetic Zucker diabetic fatty rats. AB - Although it is well established that diabetes impairs endothelium-dependent vasodilation, including those pathways involving vascular myocyte large conductance Ca(2+)-activated K(+) channels (BK(Ca)), little is known about the effects of diabetes on BK(Ca) activation as an intrinsic response to contractile stimulation. We have investigated this mechanism in a model of Type 2 diabetes, the male Zucker diabetic fatty (ZDF) rat. BK(Ca) function in prediabetic (5-7 wk) and diabetic (17-20 wk) ZDF and lean control animals was assessed in whole arteries using myograph and electrophysiology techniques and in freshly dissociated myocytes by patch clamping. Log EC(25) values for phenylephrine concentration-tension curves were shifted significantly to the left by blockade of BK(Ca) with iberiotoxin (IBTX) in arteries from non- and prediabetic animals but not from diabetic animals. Smooth muscle hyperpolarizations of arteries evoked by the BK(Ca) opener NS-1619 were significantly reduced in the diabetic group. Voltage-clamp recordings indicated that IBTX-sensitive currents were not enhanced to the extent observed in nondiabetic controls by increasing the Ca(2+) concentration in the pipette solution or the application of NS-1619 in myocytes from diabetic animals. An alteration in the expression of BK(Ca) beta(1) subunits was not evident at either the mRNA or protein level in arteries from diabetic animals. Collectively, these results suggest that myocyte BK(Ca) of diabetic animals does not significantly oppose vasoconstriction, unlike that of prediabetic and control animals. This altered function was related to a reduced Ca(2+)-dependent activation of the channel not involving beta(1) subunits. PMID- 16272201 TI - Increased natriuretic peptide receptor A and C gene expression in rats with pressure-overload cardiac hypertrophy. AB - Both atrial (ANP) and brain (BNP) natriuretic peptide affect development of cardiac hypertrophy and fibrosis via binding to natriuretic peptide receptor (NPR)-A in the heart. A putative clearance receptor, NPR-C, is believed to regulate cardiac levels of ANP and BNP. The renin-angiotensin system also affects cardiac hypertrophy and fibrosis. In this study we examined the expression of genes for the NPRs in rats with pressure-overload cardiac hypertrophy. The ANG II type 1 receptor was blocked with losartan (10 mg.kg(-1).day(-1)) to investigate a possible role of the renin-angiotensin system in regulation of natriuretic peptide and NPR gene expression. The ascending aorta was banded in 84 rats during Hypnorm/Dormicum-isoflurane anesthesia; after 4 wk the rats were randomized to treatment with losartan or placebo. The left ventricle of the heart was removed 1, 2, or 4 wk later. Aortic banding increased left ventricular expression of NPR A and NPR-C mRNA by 110% (P < 0.001) and 520% (P < 0.01), respectively, after 8 wk; as expected, it also increased the expression of ANP and BNP mRNAs. Losartan induced a slight reduction of left ventricular weight but did not affect the expression of mRNAs for the natriuretic peptides or their receptors. Although increased gene expression does not necessarily convey a higher concentration of the protein, the data suggest that pressure overload is accompanied by upregulation of not only ANP and BNP but also their receptors NPR-A and NPR-C in the left ventricle. PMID- 16272202 TI - Nitroglycerin reduces myocardial oxygen consumption during exercise despite vascular tolerance. AB - The long-term benefits of nitroglycerin (NTG) therapy are limited by the development of vascular tolerance and endothelial dysfunction in conductance coronary arteries. We have determined whether nitrate tolerance extends to NTG effects on myocardial O2 consumption (MV(O2)) and the ability of endogenous nitric oxide (NO) to modulate MV(O2) during exercise. In chronically instrumented dogs (n = 8), hemodynamic and MV(O2) responses to treadmill exercise were measured before, during tolerance (3 and 7 days of NTG delivery), and 7 days after NTG withdrawal. Acute NTG delivery caused a parallel downward shift of the MV(O2)-triple product (TP) relations and reversed the disproportionate increases in MV(O2) caused by the blockade of NO formation. After 7 days of continuous transdermal NTG delivery, vascular tolerance was displayed as a >75% reduction of coronary blood flow (CBF) responses to NTG boluses. Despite vascular nitrate tolerance, MV(O2)-TP relations were shifted downward compared with pre-NTG exercise. Seven days after NTG withdrawal, vascular responses to boluses of NTG had recovered from tolerance, and MV(O2)-TP relations during exercise were back to pre-NTG level. At that time, blockade of NO formation failed to alter MV(O2) TP relations. Thus NTG caused a sustained reduction of cardiac MV(O2), independent of metabolic demand during exercise, despite tolerance of the coronary microcirculation. NTG-induced vascular tolerance and MV(O2) reductions were reversible by NTG withdrawal, but endogenous NO-dependent modulation of O2 consumption was severely impaired. PMID- 16272203 TI - Load-sensitive measures may overestimate global systolic function in the presence of left ventricular hypertrophy: a comparison with load-insensitive measures. AB - Transgenic animal models have provided a vital insight into the pathogenesis of cardiovascular disease, but functional cardiac assessment is often limited by high heart rates and small heart size. We hypothesized that in the presence of concentric left ventricular (LV) hypertrophy (LVH), load-sensitive measures of contractility may be misinterpreted as overestimating global cardiac function, because the normal function of excess sarcomeres may displace a greater volume of blood during contraction. Conductance catheter technology was used to evaluate pressure-volume (P-V) relationships as a load-insensitive method of assessing cardiac function in vivo in 18-wk-old heterozygous (mRen-2)27 transgenic rats (a model of LVH), compared with age-matched Sprague-Dawley (SD) controls. Anesthetized animals underwent echocardiography followed by P-V loop analysis. Blood pressure, body weight, and heart rate were higher in the Ren-2 rats (P < 0.05). Load-sensitive measures of systolic function, including fractional area change, fractional shortening, ejection fraction, and positive peak rate of LV pressure development, were greater in the Ren-2 than control animals (P < 0.05). Load-insensitive measures of systolic function, including the preload recruitable stroke work relationship and the end-systolic P-V relationship, were not different between Ren-2 and SD rats. Regional wall motion assessed by circumferential shortening velocity suggested enhanced circumferential fiber contractility in the Ren-2 rats (P = 0.02), but tissue Doppler imaging, used to assess longitudinal function, was not different between groups. Although conventional measures suggested enhanced systolic function in the Ren-2 rat, load insensitive measures of contractility were not different between Ren-2 and SD animals. These findings suggest that the normal range of values for load sensitive indexes of contractility needs to be altered according to the degree of LVH. To accurately identify changes in systolic function, we suggest that a combination of echocardiography with assessment of load-insensitive measures be used routinely. PMID- 16272204 TI - Synergistic effect of angiotensin II and nitric oxide synthase inhibitor in increasing aortic stiffness in mice. AB - Although they are implicated on their own as risk factors for cardiovascular disease, the potential link between nitric oxide (NO) deficiency, ANG II, and vascular stiffening has not been tested before. We evaluated the role of chronic ANG II treatment and NO deficiency, alone and in combination, on aortic stiffness in mice and tested parameters contributing to increases in active or passive components of vascular stiffness, including blood pressure, vascular smooth muscle contractility, and extracellular matrix components. Untreated (control) mice and mice treated with a NO synthase (NOS) inhibitor [N(omega)-nitro-L arginine methyl ester (L-NAME), 0.5 g/l] were implanted with osmotic minipumps delivering ANG II (500 ng.kg(-1).min(-1)) for 28 days. Aortic stiffness was then measured in vivo by pulse wave velocity (PWV) and ex vivo by load-strain analysis to obtain values of maximal passive stiffness (MPS). Blood pressure and aortic contractility ex vivo were measured. ANG II treatment or NOS inhibition with L NAME did not independently increase vascular stiffness; however, the combined treatments worked synergistically to increase PWV and MPS. The combined treatments of ANG II + L-NAME also significantly increased aortic wall collagen content while decreasing elastin. These novel results suggest that NO deficiency and ANG II act synergistically to increase aortic stiffness in mice predominantly via changes in aortic wall collagen/elastin ratio. PMID- 16272205 TI - Absence of histamine-induced nitric oxide release in the human radial artery: implications for vasospasm of coronary artery bypass vessels. AB - Radial artery (RA) bypass grafts can develop severe vasospasm. As histamine is known to induce vasospasm, its effect on RA was assessed compared with the classic bypass vessels internal mammary artery (MA) and saphenous vein (SV). The vessels were examined in organ chambers for isometric tension recording. Histamine induced contractions on baseline; the sensitivity was higher in RA and SV than MA. After precontraction with norepinephrine, histamine did not evoke relaxations of RA but induced relaxations of MA and less of SV at lower concentrations; it induced contractions at higher concentrations, reaching similar levels in all three vessels. Indomethacin did not affect the response of MA and RA but potentiated relaxations and reduced contractions of SV. Endothelium removal, N(omega)-nitro-L-arginine methyl ester (L-NAME), or the H2-receptor blocker cimetidine did not affect the response of RA, but inhibited relaxations and enhanced contractions in MA and inhibited relaxations in SV; in the latter, only L-NAME enhanced contractions. Real-time PCR detected much lower expression of endothelial H2-receptor in RA than MA or SV. Western blots revealed similar endothelial nitric oxide (NO) synthase expression in all three vessels. Relaxations to acetylcholine were identical in RA and MA. Thus histamine releases NO by activating the endothelial H2-receptor, the expression of which is much lower in RA than MA or SV. H2-receptor activation also releases prostaglandins in SV, partially antagonizing NO. The lack of histamine-induced NO production represents a possible mechanism of RA vasospasm. PMID- 16272206 TI - In vivo evidence for endothelin-1-mediated attenuation of alpha1-adrenergic stimulation. AB - Experiments were designed to determine the influence of endothelin A (ET(A)) receptors on the pressor response to acute environmental stress in Dahl salt resistant (DR) and Dahl-sensitive (DS) rats. Mean arterial pressure (MAP) was chronically monitored by telemetry before and after treatment with the selective ET(A) receptor antagonist ABT-627. Rats were restrained and subjected to pulsatile air jet stress (3 min). In untreated animals, the total pressor response (area under the curve) to acute stress was not different between DR vs. DS rats (8.1 +/- 1.7 vs. 15.6 +/- 2.6 mmHg x 3 min, P = 0.10). Conversely, treatment with ABT-627 potentiated the total pressor response only in DR rats (36.3 +/- 6.2 vs. 22.6 +/- 5.9 mmHg x 3 min, DR vs. DS, P < 0.05). Treatment with ABT-627 allowed greater responses in anesthetized DR rats to exogenous phenylephrine (1-4 microg/kg) during ganglionic blockade (P < 0.05) and produced a significant increase in plasma norepinephrine at baseline and during stress in conscious DR rats compared with untreated animals (P < 0.05). ET(A) receptor blockade had no effect on these responses in DS rats. Our results suggest that endothelin-1 can inhibit alpha-adrenergic-mediated effects in DR, but not DS rats, consistent with the hypothesis that ET(A) receptor activation functions to reduce sympathetic nerve activity and responses in vascular smooth muscle to sympathetic stimulation. PMID- 16272207 TI - Multiple transcripts of Ca2+ channel alpha1-subunits and a novel spliced variant of the alpha1C-subunit in rat ductus arteriosus. AB - Voltage-dependent Ca(2+) channels (VDCCs), which consist of multiple subtypes, regulate vascular tone in developing arterial smooth muscle, including the ductus arteriosus (DA). First, we examined the expression of VDCC subunits in the Wistar rat DA during development. Among alpha(1)-subunits, alpha(1C) and alpha(1G) were the most predominant isoforms. Maternal administration of vitamin A significantly increased alpha(1C)- and alpha(1G)-transcripts. Second, we examined the effect of VDCC subunits on proliferation of DA smooth muscle cells. We found that 1 microM nitrendipine (an L-type Ca(2+) channel blocker) and kurtoxin (a T-type Ca(2+) channel blocker) significantly decreased [(3)H]thymidine incorporation and that 3 microM efonidipine (an L- and T-type Ca(2+) channel blocker) further decreased [(3)H]thymidine incorporation, suggesting that L- and T-type Ca(2+) channels are involved in smooth muscle cell proliferation in the DA. Third, we found that a novel alternatively spliced variant of the alpha(1C)-isoform was highly expressed in the neointimal cushion of the DA, where proliferating and migrating smooth muscle cells are abundant. The basic channel properties of the spliced variant did not differ from those of the conventional alpha(1C)-subunit. We conclude that multiple VDCC subunits were identified in the DA, and, in particular, alpha(1C)- and alpha(1G)-subunits were predominant in the DA. A novel spliced variant of the alpha(1C)-subunit gene may play a distinct role in neointimal cushion formation in the DA. PMID- 16272208 TI - Left ventricular hypertrophy in normoalbuminuric type 2 diabetic patients not taking antihypertensive treatment. AB - BACKGROUND: Left ventricular hypertrophy (LVH) is an independent risk factor for myocardial ischaemia, cardiac arrhythmia, sudden death, and heart failure, all common findings in patients with type 2 diabetes. AIM: To determine the prevalence of, and risk factors for, LVH in normoalbuminuric type 2 diabetic patients not taking antihypertensive treatment. DESIGN: Cross-sectional study. METHODS: From 1994 to 1998, M-mode echocardiography was performed by one experienced examiner in 262 consecutive, normoalbuminuric Caucasian type 2 diabetic patients, all with blood pressure <160/95 mmHg and not taking antihypertensive medication. Mean +/- SD age was 54 +/- 10 years, 109 were women, and median known duration of diabetes was 4 (range 1-28) years. Body mass index (BMI) was 28 +/- 5 kg/m(2), and blood pressure 134 +/- 13/79 +/- 8 mmHg, all means +/- SD. Median urinary albumin excretion rate was 9 (range 2-30) mg/24 h. RESULTS: The prevalence of LVH indexed to height(2.7) was 43% (95%CI 38-50%), and was similar in men and women. BMI, HbA(1c) and log urinary albumin excretion were significantly associated with left ventricular hypertrophy in a logistic regression model, whereas sex, age, known duration of diabetes and blood pressure were not. Similar results were obtained for left ventricular mass index. DISCUSSION: LVH was frequent in our normoalbuminuric type 2 diabetic patients not taking antihypertensive treatment. Several potentially modifiable risk factors, such as raised BMI, poor glycaemic control and elevated urinary albumin excretion rate, were associated with LVH. PMID- 16272209 TI - Masticatory performance in children and adolescents with Class I and II malocclusions. AB - It is not fully understood whether masticatory performance is compromised in individuals with the more common forms of malocclusion (i.e. Class I and Class II). The aim of this prospective investigation was to establish the relationships between masticatory performance, malocclusion (type and severity), age, body size and gender, in children and adolescents. A total of 335 individuals were examined at the average ages of 6, 9, 12 and 15 years. Each subject's occlusal status was described by Angle classification and by the Peer Assessment Ratio (PAR) index. Masticatory performance was quantified by the median particle size (MPS) and the broadness of particle distribution using artificial food. Masticatory performance improved significantly with age. The 6-year-old children were less able to break down the food particles (MPS 4.20 mm2) than the 15 year olds (MPS 3.24 mm2). Analysis of covariance showed that age differences in performance are related to an increase in body size. There were statistically significant differences in masticatory performance between children with normal occlusion and those with a Class I malocclusion; no differences were found between normal occlusion and Class II malocclusion. Gender differences did not explain the variation in masticatory performance. It is concluded that occlusal indices are not reliable predictors of masticatory performance. Traditional descriptors of malocclusion type and severity apparently cannot explain most of the variation in masticatory performance in children and adolescents. PMID- 16272210 TI - A new management of syncope: prospective systematic guideline-based evaluation of patients referred urgently to general hospitals. AB - AIMS: The guidelines of the European Society of Cardiology (ESC) define the current standard for the management of syncope, but are still incompletely applied in the clinical setting. METHODS AND RESULTS: Prospective systematic evaluation, on strict adherence to the guidelines, of consecutive patients referred for syncope to the emergency departments of 11 general hospitals. In order to maximize the application, a decision-making guideline-based software was used and trained core medical personnel were designated-both locally in each hospital and centrally-to verify adherence to the diagnostic pathway and give advice on its correction. A diagnostic work-up consistent with the guidelines was completed in 465/541 patients (86%). A definite diagnosis was established in 98% (unexplained in 2%): neurally mediated syncope accounted for 66% of diagnosis, orthostatic hypotension 10%, primary arrhythmias 11%, structural cardiac or cardiopulmonary disease 5%, and non-syncopal attacks 6%, respectively. The initial evaluation (consisting of history, physical examination, and standard electrocardiogram) established a diagnosis in 50% of cases. Hospitalization for the management of syncope was appropriate in 25% and was required for other reasons in a further 13% of cases. The median in-hospital stay was 5.5 days (interquartile range, 3-9). Apart from the initial evaluation, a mean of 1.9+/ 1.1 appropriate tests per patient was performed in 193 patients and led to a final diagnosis in 182 of these (94%). CONCLUSION: The results of this study assess the current standard for the management of syncope on the basis of a rigorous adherence to guidelines of the ESC and provide a frame of reference for daily activity when dealing with syncope. PMID- 16272211 TI - Predictors of revascularization method and long-term outcome of percutaneous coronary intervention or repeat coronary bypass surgery in patients with multivessel coronary disease and previous coronary bypass surgery. AB - AIMS: The optimal revascularization strategy in patients with symptomatic multivessel coronary artery disease (CAD) and previous coronary artery bypass grafting (CABG) remains unknown. METHODS AND RESULTS: We evaluated all patients with previous CABG undergoing isolated, non-emergency multivessel revascularization between 1 January 1995 and 31 December 2000. The analysis concentrated on the independent predictors of the revascularization method, as well as on long-term mortality and its predictors, after calculating a propensity score for the method of revascularization. There were 2191 patients (1487 with reoperation and 704 with percutaneous coronary intervention, PCI) in the study. The most important factors in choosing reoperation were presence of more diseased or occluded grafts, previous infarction, lower ejection fraction (EF), longer interval from first CABG, and more total occlusions of native arteries, as well as absence of a patent mammary graft. The distribution of the propensity score was skewed towards the two extremes. At 5 years, the unadjusted cumulative survival was 79.5% for CABG and 75.3% for PCI, P=0.008. After adjustment for the propensity score for PCI vs. CABG, PCI was associated with a hazard ratio of 1.47 (0.94-2.28), P=0.09. The most powerful predictors of mortality were higher age and lower EF. CONCLUSION: The choice of the revascularization method in patients with previous CABG is dictated mostly by anatomical considerations and less by clinical characteristics. In contrast, clinical characteristics predominantly affect long-term outcome, whereas the method of revascularization has a limited effect. A randomized clinical trial addressing this important segment of the population with ischaemic heart disease is warranted. PMID- 16272212 TI - Is the level of evidence for the use of beta-blockers in acute myocardial infarction satisfactory enough? PMID- 16272213 TI - Matrix metalloproteinases and cardiovascular disease. PMID- 16272214 TI - Eating, vascular biology, and atherosclerosis: a lot to chew on. PMID- 16272215 TI - Syncope Management Guidelines at work: first steps towards assessing clinical utility. PMID- 16272216 TI - Demonstration by heterologous expression that the Leishmania SCA1 gene encodes an arabinopyranosyltransferase. AB - In part of the life cycle within their sand fly vector, Leishmania major parasites first attach to the fly's midgut through their main surface adhesin lipophosphoglycan (LPG) and later resynthesize a structurally distinct LPG that results in detachment and eventual transmission. One of these structural modifications requires the addition of alpha1,2-D-arabinopyranose caps to beta1,3 galactose side chains in the phosphoglycan repeat unit domain of LPG. We had previously identified two side chain arabinose genes (SCA1/2) that were involved in the alpha1,2-D-Arap capping. SCA1/2 exhibit canonical glycosyltransferase motifs, and overexpression of either gene leads to elevated microsomal alpha1,2-D ArapT activity, resulting in arabinopyranosylation of beta1,3-Gal side chains in LPG (hereafter called side chain D-arabinopyranosyltransferase [sc-D-ArapT]). Heterologous expression in a null arabinose background was used to determine whether the SCA1 gene encodes the actual sc-D-ArapT. SCA1 expression constructs introduced into both mammalian COS-7 cells and the baculovirus-sf9 cell system exhibited considerable expression of the protein. However, functional sc-D-ArapT activity was observed only in the latter. In in vitro assays incubated with guanidine 59-diphosphate (GDP)-D-[3H]Arap as the sugar donor and utilizing exogenous LPG as an acceptor, significant sc-D-ArapT activity was observed when microsomes from the baculovirus-sf9 cells were incubated in presence of the LPG acceptor. No activity was observed in the absence of LPG. These results demonstrate that SCA1 encodes a sc-D-ArapT and provide the first example of heterologous expression of a D-ArapT gene. PMID- 16272217 TI - The delta subunit of gamma-aminobutyric acid type A receptors does not confer sensitivity to low concentrations of ethanol. AB - GABA(A) receptors (GABA(A)Rs) are usually formed by alpha, beta, and gamma or delta subunits. Recently, delta-containing GABA(A)Rs expressed in Xenopus oocytes were found to be sensitive to low concentrations of ethanol (1-3 mM). Our objective was to replicate and extend the study of the effect of ethanol on the function of alpha4beta3delta GABA(A)Rs. We independently conducted three studies in two systems: rat and human GABA(A)Rs expressed in Xenopus oocytes, studied through two-electrode voltage clamp; and human GABA(A)Rs stably expressed in the fibroblast L(tk-) cell line, studied through patch-clamp electrophysiology. In all cases, alpha4beta3delta GABA(A)Rs were only sensitive to high concentrations of ethanol (100 mM in oocytes, 300 mM in the cell line). Expression of the delta subunit in oocytes was assessed through the magnitude of the maximal GABA currents and sensitivity to zinc. Of the three rat combinations studied, alpha4beta3 was the most sensitive to ethanol, isoflurane, and 5alpha-pregnan 3alpha,21-diol-20-one (THDOC); alpha4beta3delta and alpha4beta3gamma(2S) were very similar in most aspects, but alpha4beta3delta was more sensitive to GABA, THDOC, and lanthanum than alpha4beta3gamma(2S) GABA(A)Rs. Ethanol at 30 mM did not affect tonic GABA-mediated currents in dentate gyrus reported to be mediated by GABA(A)Rs incorporating alpha4 and delta subunits. We have not been able to replicate the sensitivity of alpha4beta3delta GABA(A)Rs to low concentrations of ethanol in four different laboratories in independent studies. This suggests that as yet unidentified factors may play a critical role in the ethanol effects on delta-containing GABA(A)Rs. PMID- 16272218 TI - Pharmacological characterization of agonists at delta-containing GABAA receptors: Functional selectivity for extrasynaptic receptors is dependent on the absence of gamma2. AB - Several groups have characterized the pharmacology of alpha4- or alpha6beta3delta containing GABA(A) receptors expressed in different cell systems. We have previously demonstrated that the pharmacological profiles of a series of GABA(A) receptor agonists are highly dependent on the alpha subunit and little on the beta and gamma subunits, so to further understand the contribution of the different subunits in the GABA(A) receptor complex, we characterized a series of full agonists, partial agonists, and antagonists at alpha4beta3, alpha4beta3delta, and alpha6beta3delta receptors expressed in Xenopus oocytes. Little or no difference was seen when the compounds were compared at alphabeta- and alphabetadelta-containing receptors, whereas a significant reduction in both potency and relative efficacy was observed compared with alphabetagamma containing receptors described in the literature. These data clearly confirm that the presence of the delta subunit in heterotrimeric receptors is a strong determinant of the increased pharmacological activity of compounds with agonist activity. The very similar agonist pharmacology of alphabeta- and alphabetadelta containing receptors, which is significantly different from that of alphabetagamma-containing receptors, shows that whereas the presence of a gamma subunit impairs the response to an agonist stimulation of the alphabeta receptor complex, the delta subunit does not affect this in any way. Taken together, these data are well in line with the idea that alpha4beta3delta may contribute to the pharmacological action of exogenously applied agonists and may explain why systemically active compounds such as gaboxadol and muscimol in vivo appear to act as selective extrasynaptic GABA(A) agonists. PMID- 16272219 TI - Revealing the large-scale network organization of growth hormone-secreting cells. AB - Pituitary growth hormone (GH)-secreting cells regulate growth and metabolism in animals and humans. To secrete highly ordered GH pulses (up to 1,000-fold rise in hormone levels in vivo), the pituitary GH cell population needs to mount coordinated responses to GH secretagogues, yet GH cells display an apparently heterogeneous scattered distribution in 2D histological studies. To address this paradox, we analyzed in 3D both positioning and signaling of GH cells using reconstructive, two-photon excitation microscopy to image the entire pituitary in GH-EGFP transgenic mice. Our results unveiled a homologous continuum of GH cells connected by adherens junctions that wired the whole gland and exhibited the three primary features of biological networks: robustness of architecture across lifespan, modularity correlated with pituitary GH contents and body growth, and connectivity with spatially stereotyped motifs of cell synchronization coordinating cell activity. These findings change our view of GH cells, from a collection of dispersed cells to a geometrically connected homotypic network of cells whose local morphology and connectivity can vary, to alter the timing of cellular responses to promote more coordinated pulsatile secretion. This large scale 3D view of cell functioning provides a powerful approach to identify and understand other networks of endocrine cells that are thought to be scattered in situ. Many dispersed endocrine systems exhibit pulsatile outputs. We suggest that cell positioning and associated cell-cell connection mechanisms will be critical parameters that determine how well such systems can deliver a coordinated secretory pulse of hormone to their target tissues. PMID- 16272220 TI - A yeast two-hybrid knockout strain to explore thioredoxin-interacting proteins in vivo. AB - All organisms contain thioredoxin (TRX), a regulatory thiol:disulfide protein that reduces disulfide bonds in target proteins. Unlike animals and yeast, plants contain numerous TRXs for which no function has been assigned in vivo. Recent in vitro proteomic approaches have opened the way to the identification of >100 TRX putative targets, but of which none of the numerous plant TRXs can be specifically associated. In contrast, in vivo methodologies, including classical yeast two-hybrid (Y2H) systems, failed to reveal the expected high number of TRX targets. Here, we developed a yeast strain named CY306 designed to identify TRX targets in vivo by a Y2H approach. CY306 contains a GAL4 reporter system but also carries deletions of endogenous genes encoding cytosolic TRXs (TRX1 and TRX2) that presumably compete with TRXs introduced as bait. We demonstrate here that, in the CY306 strain, yeast TRX1 and TRX2, as well as Arabidopsis TRX introduced as bait, interact with known TRX targets or putative partners such as yeast peroxiredoxins AHP1 and TSA1, whereas the same interactions cannot be detected in classical Y2H strains. Thanks to CY306, we also show that TRXs interact with the phosphoadenosine-5-phosphosulfate (PAPS) reductase MET16 through a conserved cysteine. Moreover, interactions visualized in CY306 are highly specific depending on the TRX and targets tested. CY306 constitutes a relevant genetic system to explore the TRX interactome in vivo and with high specificity, and opens new perspectives in the search for new TRX-interacting proteins by Y2H library screening in organisms with multiple TRXs. PMID- 16272222 TI - Bees encode behaviorally significant spectral relationships in complex scenes to resolve stimulus ambiguity. AB - Bees, like humans, can continue to see a surface from its color even when the scene's global illuminant changes (which is a phenomenon called color constancy). It is not known, however, whether they can also generate color-constant behavior in more natural complex scenes that are lit by multiple lights simultaneously, conditions in which most computational models of color constancy fail. To test whether they can indeed solve this more complex problem, bumblebees were raised in a highly controlled, yet ecological relevant environment consisting of a matrix of 64 artificial flowers under four spatially distinct lights. As in nature, the bees had no direct access to spectral information about the illuminants or flowers. Furthermore, the background of all of the flowers in the matrix was black, independent of illumination. The stimulus information presented to the bee was, therefore, far more constrained than that normally experienced in nature. And yet, bees learned to identify the rewarded flowers in each differently illuminated region of the matrix, even when the illumination of one of the regions was switched with one the bees had not previously experienced. These results suggest that bees can generate color-constant behavior by encoding empirically significant contrast relationships between statistically dependent, but visually distinct, stimulus elements of scenes. PMID- 16272221 TI - Inhibition of FLT3 signaling targets DCs to ameliorate autoimmune disease. AB - Autoimmune diseases often result from inappropriate or unregulated activation of autoreactive T cells. Traditional approaches to treatment of autoimmune diseases through immunosuppression have focused on direct inhibition of T cells. In the present study, we examined the targeted inhibition of antigen-presenting cells as a means to downregulate immune responses and treat autoimmune disease. Dendritic cells (DCs) are the central antigen-presenting cells for the initiation of T cell responses, including autoreactive ones. A large portion of DCs are derived from hematopoietic progenitors that express FLT3 receptor (CD135), and stimulation of the receptor via FLT3 ligand either in vivo or in vitro is known to drive expansion and differentiation of these progenitors toward a DC phenotype. We hypothesized that inhibition of FLT3 signaling would thus produce an inhibition of DC-induced stimulation of T cells, thereby inhibiting autoimmune responses. To this end, we used small-molecule tyrosine kinase inhibitors targeted against FLT3 and examined the effects on DCs and their role in the promulgation of autoimmune disease. Results of our studies show that inhibition of FLT3 signaling induces apoptosis in both mouse and human DCs, and thus is a potential target for immune suppression. Furthermore, targeted inhibition of FLT3 significantly improved the course of established disease in a model for multiple sclerosis, experimental autoimmune encephalomyelitis, suggesting a potential avenue for treating autoimmune disease. PMID- 16272223 TI - Differential use of two cyclic electron flows around photosystem I for driving CO2-concentration mechanism in C4 photosynthesis. AB - Whereas linear electron flow (LEF) in photosynthesis produces both ATP and NADPH, the cyclic electron flow (CEF) around photosystem I has been shown to produce only ATP. Two alternative routes have been shown for CEF; NAD(P)H dehydrogenase (NDH)- and ferredoxin:plastoquinone oxidoreductase (FQR)-dependent flows, but their physiological relevance has not been elucidated in detail. Meanwhile, because C(4) photosynthesis requires more ATP than does C(3) photosynthesis to concentrate CO(2), it has not been clear how the extra ATP is produced. In this study, to elucidate whether CEF contributes to the additional ATP needed in C(4) photosynthesis, we estimated the amounts of PGR5, which participates in FQR dependent flow, and NDH-H, a subunit of NDH, in four C(4) species. Although the expression profiles of PGR5 did not correlate well with the additional ATP requirement, NDH was greatly expressed in mesophyll cells in the NAD-malic enzyme (ME) species, and in bundle-sheath cells in NADP-ME species, where there is a strong need for ATP in the respective cells. Our results indicate that CEF via NDH plays a central role in driving the CO(2)-concentrating mechanism in C(4) photosynthesis. PMID- 16272224 TI - Pharmacological properties of GABAA receptors containing gamma1 subunits. AB - GABA(A) receptors composed of alpha(1), beta(2), gamma(1) subunits are expressed in only a few areas of the brain and thus represent interesting drug targets. The pharmacological properties of this receptor subtype, however, are largely unknown. In the present study, we expressed alpha(1)beta(2)gamma(1)-GABA(A) receptors in Xenopus laevis oocytes and analyzed their modulation by 21 ligands from 12 structural classes making use of the two-microelectrode voltage-clamp method and a fast perfusion system. Modulation of GABA-induced chloride currents (I(GABA)) was studied at GABA concentrations eliciting 5 to 10% of the maximal response. Triazolam, clotiazepam, midazolam, 2-(4-methoxyphenyl)-2,3,5,6,7,8,9,10 octahydro-cyclohepta-(b)pyrazolo[4,3-d]pyridin-3-one (CGS 20625), 2-(4 chlorophenyl)-pyrazolo[4,3-c]quinolin-3-one (CGS 9896), diazepam, zolpidem, and bretazenil at 1 microM concentrations were able to significantly (>20%) enhance I(GABA) in alpha(1)beta(2)gamma(1) receptors. Methyl-6,7-dimethoxy-4-ethyl-beta carboline-3-carboxylate, 3-methyl-6-[3-trifluoromethyl-phenyl]-1,2,4-triazolo[4,3 b]pyridazine (Cl 218,872), clobazam, flumazenil, 5-(6-ethyl-7-methoxy-5 methylimidazo[1,2-a]pyrimidin-2-yl)-3-methyl-[1,2,4]-oxadiazole (Ru 33203), 2 phenyl-4-(3-ethyl-piperidinyl)-quinoline (PK 9084), flurazepam, ethyl-7-methoxy 11,12,13,13a-tetrahydro-9-oxo-9H-imidazo[1,5-a]pyrrolo[2,1-c] [1,4]benzodiazepine 1-carboxylate (l-655,708), 2-(6-ethyl-7-methoxy-5-methylimidazo[1,2-a]pyrimidin-2 yl)-4-methyl-thiazole (Ru 33356), and 6-ethyl-7-methoxy-5-methylimidazo[1,2 a]pyrimidin-2-yl)phenylmethanone (Ru 32698) (1 microM each) had no significant effect, and flunitrazepam and 2-phenyl-4-(4-ethyl-piperidinyl)-quinoline (PK 8165) inhibited I(GABA). The most potent compounds triazolam, clotiazepam, midazolam, and CGS 20625 were investigated in more detail on alpha(1)beta(2)gamma(1) and alpha(1)beta(2)gamma(2S) receptors. The potency and efficiency of these compounds for modulating I(GABA) was smaller for alpha(1)beta(2)gamma(1) than for alpha(1)beta(2)gamma(2S) receptors, and their effects on alpha(1)beta(2)gamma(1) could not be blocked by flumazenil. CGS 20625 displayed the highest efficiency by enhancing at 100 microM I(GABA) (alpha(1)beta(2)gamma(2)) by 775 +/- 17% versus 526 +/- 14% I(GABA) (alpha(1)beta(2)gamma(1)) and 157 +/- 17% I(GABA) (alpha(1)beta(2)) (p < 0.05). These data provide new insight into the pharmacological properties of GABA(A) receptors containing gamma(1) subunits and may aid in the design of specific ligands for this receptor subtype. PMID- 16272225 TI - Sperm transport in the female reproductive tract. AB - At coitus, human sperm are deposited into the anterior vagina, where, to avoid vaginal acid and immune responses, they quickly contact cervical mucus and enter the cervix. Cervical mucus filters out sperm with poor morphology and motility and as such only a minority of ejaculated sperm actually enter the cervix. In the uterus, muscular contractions may enhance passage of sperm through the uterine cavity. A few thousand sperm swim through the uterotubal junctions to reach the Fallopian tubes (uterine tubes, oviducts) where sperm are stored in a reservoir, or at least maintained in a fertile state, by interacting with endosalpingeal (oviductal) epithelium. As the time of ovulation approaches, sperm become capacitated and hyperactivated, which enables them to proceed towards the tubal ampulla. Sperm may be guided to the oocyte by a combination of thermotaxis and chemotaxis. Motility hyperactivation assists sperm in penetrating mucus in the tubes and the cumulus oophorus and zona pellucida of the oocyte, so that they may finally fuse with the oocyte plasma membrane. Knowledge of the biology of sperm transport can inspire improvements in artificial insemination, IVF, the diagnosis of infertility and the development of contraceptives. PMID- 16272226 TI - Is the paraneoplastic syndrome of inappropriate antidiuretic hormone secretion in lung cancer always attributable to the small cell variety? AB - The syndrome of inappropriate antidiuretic hormone (SIADH) secretion is a well recognised paraneoplastic phenomenon and the vast majority are associated with small cell lung carcinoma. Rarely however, the non-small cell variety can sometimes be responsible and this report describes such an occurrence. Uniquely in this case, after chemotherapy the paraneoplastic SIADH improved in parallel with a tumour response and this has not been reported previously. PMID- 16272227 TI - A rare cause of mediastinal expansion with a massive pleural effusion. AB - The authors present a case of a 53 year old woman, who was admitted to hospital because of an unusual cause of massive pleural effusion. During diagnostic examination the mediastinal propagation of the pancreatic pseudocyst was discovered as a complication of the chronic calcifying pancreatitis. The patient was operated on and the pseudocyst was resolved by Roux-en-Y cystjejunostomy. The diagnostics and treatment of this unusual pancreatic pseudocyst spreading is discussed. PMID- 16272229 TI - Access and equity to cancer care in the USA: a review and assessment. AB - Cancer represents a serious threat to the health of women and men living in the USA. As the second leading cause of death, it claims about 500,000 lives annually. Health disparities occur when there are differences in the incidence, prevalence, mortality, and burden of disease among specific sub-populations within a specified region. For decades, disparities have been reported among Americans from racial/ethnic minority groups and those from low income groups. African Americans, the largest racial minority group in the USA, have the highest cancer incidence and mortality rates in the USA; it is about 10% higher in African Americans than in white people. Inequities in insurance status among Americans adversely affect their ability to obtain the entire range of cancer care. Those who are members of ethnic minorities and the working poor are especially apt to have poorer access to care and reduced quality of cancer care services as a result. PMID- 16272230 TI - Is antibiotic resistance a problem? A practical guide for hospital clinicians. AB - Antibiotic resistance is an important concern for patients, physicians, healthcare managers, and policymakers. Inappropriate antimicrobial prescribing fuels the evolution of resistance, while poor basic hygiene facilitates the spread of resistant microbes between patients and healthcare staff. The development of infection with a resistant pathogen may lead to poorer health and economic outcomes. The problem for the frontline clinician, however, is how to balance the responsibility of prudent prescribing with the risk of sub-optimally treating a patient who may be infected with a resistant pathogen. This article discusses how hospital physicians can use severity and risk factor assessment, and knowledge of local microbial epidemiology, to guide empiric antibiotic prescribing. Most patients hospitalised with a community acquired bacterial infection in the UK can still be managed with a traditional first line antibiotic(s). In contrast, regimens that account for resistance are often required in patients with hospital acquired infections, particularly if the patient is critically ill. PMID- 16272231 TI - Food allergy in children. AB - Food allergy is being increasingly recognised with the highest prevalence being in preschool children. Pathogenesis varies so diagnosis rests on careful history and clinical examination, appropriate use of skin prick and serum-specific IgE testing, food challenge, and supervised elimination diets. A double blind placebo controlled food challenge is the gold standard diagnostic test. Avoidance of the allergenic food is the key towards successful management. IgE mediated food allergy may present as a potentially fatal anaphylactic reaction, and management consists of the appropriate use of adrenaline (epinephrine) and supportive measures. Sensitisation remains a key target for intervention. Disease modifying agents are currently under trial for managing difficult allergies. Management requires a multidisciplinary approach and follow up. PMID- 16272234 TI - Branch retinal artery occlusion during coronary angiography. PMID- 16272236 TI - Epidemiology, clinical characteristics, and management of adults referred to a teaching hospital first seizure clinic. AB - BACKGROUND: There are scarce data describing the epidemiology, clinical characteristics, and management of adults who suffer a suspected first seizure. AIM: To describe the epidemiology, clinical characteristics, and management of adults with a suspected first seizure who are referred to a teaching hospital first seizure clinic over a one year period. DESIGN: Prospective descriptive study. METHODS: Data were collected on consecutive adults referred to the Royal Infirmary of Edinburgh between 4 February 2003 and 10 February 2004. RESULTS: 232 patients were referred to the first seizure clinic. Median age was 32 years; 53% of patients were male. Lower socioeconomic groups were more likely to present with a suspected first seizure. Nineteen per cent of patients were admitted to hospital after their suspected seizure episode. Appropriate driving advice was reported in 64% of cases. Seventy two per cent of patients were offered a first seizure clinic appointment within six weeks of referral. Nine per cent of patients had a subsequent seizure while awaiting review. Fifty two per cent of patients were confirmed as having a first seizure at the clinic, of which 56% were provoked by alcohol, recreational drugs, or sleep deprivation. Electroencephalography and computed tomography of the brain were the most common investigations ordered at the first seizure clinic (22% and 22% of patients respectively). CONCLUSION: Adults who suffer a suspected first seizure, and who make a full neurological recovery, can be safely managed as an outpatient. Around half of these patients will have a specialist diagnosis of first seizure and alcohol will be a common precipitating factor. PMID- 16272235 TI - Systolic and diastolic function in middle aged patients with sickle beta thalassaemia. An echocardiographic study. AB - OBJECTIVE: To evaluate the right and left ventricular systolic and diastolic function in middle aged patients with sickle beta thalassaemia. METHODS: Forty three patients with sickle beta thalassaemia were recruited for echocardiographic study while 55 controls, matched for age and sex, served as the control group. Parameters measured included: dimensions and wall thickness of left (LV) and right (RV) ventricle and left atrium, LV mass, and cardiac index. LV and RV contractility variables--ejection fraction, circumferential fibre shortening velocity, end systolic stress, end systolic stress/volume index ratio, mitral and tricuspid annulus systolic excursion, and Tei index--were also calculated. The study also evaluated parameters of RV and LV diastolic function including early and late atrioventricular flow velocities (E and A wave respectively), E/A ratio, deceleration time (DT), isovolumic relaxation time (IVRT) as well as pulmonary and hepatic veins systolic to diastolic (S/D) ratio. RESULTS: Chamber enlargement, greater LV mass index, cardiac index, and RV wall thickness were found in the anaemic group compared with controls. The LV and RV contractility variables of the patients were similar to controls. Conversely the LV and RV Tei index was significantly greater in the patient group. Diastolic dysfunction was present in the anaemic patients resulting from the increased LV and RV A-wave, the longer LVIVRT, RVIVRT, and RVDT, as well as the higher hepatic and pulmonary veins S/D ratio. CONCLUSIONS: The results show that in middle aged patients with sickle beta thalassaemia the diastolic function is abnormal in both ventricles but still more in RV, whereas the systolic function remains unchanged. PMID- 16272233 TI - Pleural effusion. AB - Pleural disease remains a commonly encountered clinical problem for both general physicians and chest specialists. This review focuses on the investigation of undiagnosed pleural effusions and the management of malignant and parapneumonic effusions. New developments in this area are also discussed at the end of the review. It aims to be evidence based together with some practical suggestions for practising clinicians. PMID- 16272237 TI - Pragmatic, observational study of bupropion treatment for smoking cessation in general practice. AB - BACKGROUND: Cigarette smoking remains the single largest cause of premature death in the United Kingdom. As part of the government's national service framework for coronary heart disease, smoking cessation forms a key part of the strategy. OBJECTIVES: To determine the effectiveness of bupropion treatment for smoking cessation in a general practice setting, measuring continuous abstinence from smoking, from 8 weeks to 52 weeks. DESIGN: Prospective observational study. SETTING: One general practice (six whole time equivalent doctors, 11,070 patients) in rural Northumberland. SUBJECTS: Of the 243 patients who presented to the practice over a one year period for smoking cessation, a total of 227 motivated people, who were appropriate for bupropion treatment as a pharmacological aid for smoking cessation, entered the study. Continuous smoking cessation at one year was validated by an exhaled carbon monoxide level of 10 ppm or less. RESULTS: Fifty patients successfully gave up smoking, giving a one year smoking cessation prevalence with bupropion of 22% (95% confidence intervals (CI) 17% to 28%). There was no difference in success rate for sex, number of cigarettes smoked, the number of years smoking, or whether there were other smokers in the household or not. CONCLUSION: Bupropion treatment in this general practice helped 22% of motivated people to quit and remain stopped smoking at one year. Mainly nurses, whose prescribing rights are restricted and currently exclude bupropion, deliver smoking cessation services in primary care. PMID- 16272238 TI - Clinical [corrected] and pathological characteristics of Chinese patients with antineutrophil cytoplasmic autoantibody associated systemic vasculitides: a study of 426 patients from a single centre. AB - BACKGROUND: Antineutrophil cytoplasmic autoantibodies (ANCA) are serological markers of ANCA associated systemic vasculitides (AASV), which is one of the most common multisystem autoimmune diseases. Features of Chinese patients with AASV have not been fully investigated. OBJECTIVE: To analyse the clinical and pathological characteristics of Chinese patients with AASV. METHODS: 426 Chinese patients with AASV diagnosed in the past eight years were retrospectively studied and their clinical and pathological data were analysed. RESULTS: Of the 426 patients, 87 (20.4%) were Wegener's granulomatosis, 337 (79.1%) were microscopic polyangiitis and two (0.5%) were Churg-Strauss syndrome. Only 201 of 426 (47.2%) patients were diagnosed within three months. Clinically, the patients had multisystem involvement. Altogether 371 of 426 (87.1%) had kidney involvement and 260 of 426 (61.0%) had lung involvement. The prevalences of renal involvement and fatigue were significantly higher in patients with MPO-ANCA than that in patients with PR3-ANCA; the prevalences of ophthalmic, nasal involvement, rash, and arthragia were significantly higher in patients with PR3-ANCA than those in patients with MPO-ANCA. The one and five year death rates were 13.1% and 22.4%, respectively. The percentage of patients progressing to end stage renal disease at one and five years was 15.9% and 27.1%, respectively. CONCLUSIONS: AASV is not a rare autoimmune disease in Chinese people. Kidney and lung were the most vulnerable organs. For patients with multiorgan damage, an ANCA test should be performed to make an early diagnosis and to start treatment in time. PMID- 16272239 TI - Endoscopy waiting times and impact of the two week wait scheme on diagnosis and outcome of upper gastrointestinal cancer. AB - The NHS has introduced the two week wait scheme to detect upper gastrointestinal cancers at an early stage and improve survival rates The aim of this study was to assess the impact of this scheme and changes in endoscopy waiting times on tumour stage and resection rates over a four year period. Data were analysed prospectively for all patients diagnosed with oesophagogastric cancer between September 1998 and September 2002 and from those referred under the two week wait scheme since its introduction in 2000. Of those tumours diagnosed by this scheme (15%) only 5% were early disease (stage 1 or 2). Patients with early cancer, mainly diagnosed by routine gastroscopy, do not present with symptoms meeting the two week wait criteria. An increase in the resection rates for early disease will most probably be seen with a reduction in routine endoscopy waiting times. PMID- 16272240 TI - The Dynamics of Quadrupedal Walking. 1938 - A step forward for locomotor mechanics. PMID- 16272241 TI - Muscle biochemistry and the ontogeny of flight capacity during behavioral development in the honey bee, Apis mellifera. AB - A fundamental issue in physiology and behavior is underlie major behavioral shifts in organisms as they transitions are common in nature and include the age related switch from nest/hive work to foraging in social insects such as honey bees (understanding the functional and genetic mechanisms that adopt new environments or life history tactics. Such). Because of their experimental Apis mellifera tractability, recently sequenced genome and well understood biology, honey bees are an ideal model system for integrating molecular, genetic, physiological and sociobiological perspectives to advance understanding of behavioral and life history transitions. When honey bees (Apis mellifera) transition from hive work to foraging, their flight muscles undergo changes Apis mellifera that allow these insects to attain the highest rates of flight muscle metabolism and power output ever recorded in the animal kingdom. Here, we review research to date showing that honey bee flight muscles undergo significant changes in biochemistry and gene expression and that these changes accompany a significant increase in the capacity to generate metabolic and aerodynamic power during flight. It is likely that changes in muscle gene expression, biochemistry, metabolism and functional capacity may be driven primarily by behavior as opposed to age, as is the case for changes in honey bee brains. PMID- 16272242 TI - Switch to diester preen waxes may reduce avian nest predation by mammalian predators using olfactory cues. AB - It has long been recognised that nest depredation by olfactory-searching mammals greatly influences the reproductive success of ground-nesting birds. Yet adaptations of birds to diminish smell during nesting have rarely been investigated. Recently, a remarkable shift in the composition of uropygial gland secretions (preen waxes) was discovered in many ground-nesting shorebirds and ducks that begin incubation, during which the usual mixtures of monoester preen waxes are replaced by mixtures of less volatile diester waxes. In this study we show experimentally that an olfactory-searching dog had greater difficulty detecting mixtures of the less volatile diesters than mixtures of monoesters. This is consistent with the hypothesis that diester preen waxes reduce birds' smell and thereby reduce predation risk. PMID- 16272244 TI - Response properties of electrosensory afferent fibers and secondary brain stem neurons in the paddlefish. AB - The passive electrosense is used by many aquatic animals to detect weak electric fields from other animals or from geoelectric sources. In contrast to the active electrosense, ;passive' means that there are no electric organs, and only external fields are measured. Electroreceptors are distributed in the skin, but are different from other skin senses because they can detect and localize sources a considerable distance away. Distant sources, however, stimulate a large number of receptors at the same time and central circuits have to compute the exact location of the source from this distributed information. In order to gain insights into the algorithms involved, we compared the response properties of units in the dorsal octavolateral nucleus (DON) with primary afferent fibers in the paddlefish. The following parameters were tested: spontaneous activity, sensitivity, frequency tuning, receptive field size, movement sensitivity, and topography within the DON. Although there are some differences in spontaneous activity and receptive field size, there are no major differences between primary afferents and DON units that could reveal any substantial amount of spatial information processing. In particular the lack of any topographic order within the DON renders a lateral interaction between neighboring receptive fields unlikely. PMID- 16272243 TI - Expression and functional analysis of mussel taurine transporter, as a key molecule in cellular osmoconforming. AB - Most aquatic invertebrates adapt to environmental osmotic changes primarily by the cellular osmoconforming process, in which osmolytes accumulated in their cells play an essential role. Taurine is one of the most widely utilized osmolytes and the most abundant in many molluscs. Here, we report the structure, function and expression of the taurine transporter in the Mediterranean blue mussel (muTAUT), as a key molecule in the cellular osmoconforming process. Deduced amino acid sequence identity among muTAUT and vertebrate taurine transporters is lower (47-51%) than that among vertebrate taurine transporters (>78%). muTAUT has a lower affinity and specificity for taurine and a requirement for higher NaCl concentration than vertebrate taurine transporters. This seems to reflect the internal environment of the mussel; higher NaCl and taurine concentrations. In addition to the hyperosmotic induction that has been reported for cloned taurine transporters, the increase in muTAUT mRNA was unexpectedly observed under hypoosmolality, which was depressed by the addition of taurine to ambient seawater. In view of the decrease in taurine content in mussel tissue under conditions of hypoosmolality reported previously, our results lead to the conclusion that muTAUT does not respond directly to hypoosmolality, but to the consequent decrease in taurine content. By immunohistochemistry, intensive expression of muTAUT was observed in the gill and epithelium of the mantle, which were directly exposed to intensive osmotic changes of ambient seawater. PMID- 16272245 TI - Desert ants compensate for navigation uncertainty. AB - During foraging trips, desert ants Cataglyphis fortis do not rely only on their well-studied path integration system, they also use olfactory cues when approaching a familiar food source. When a wind is blowing from a constant direction, as is characteristic of their desert habitat, the ants do not approach the feeder directly. They rather steer some distance downwind of the food source to pick up odour filaments emanating from the food. They follow this odour trail upwind, and find the source quickly and reliably. This approach behaviour was examined in more detail in order to identify the underlying orientation strategy. First, the ants may employ a 'goal expansion strategy', using odour spread as a spatially limited indicator for the presence of food. In that case, the distance steered downwind of the feeder should be determined by the range of the odour plume (and, for instance, wind speed). It should be independent of the distance between nest and feeder. Second, the ants may apply an 'error compensation strategy', using odour filaments as a guideline towards the food source. Steering downwind by a margin just exceeding their maximum navigation error will lead the ants safely across the odour guide. In that case, the distance steered downwind of the feeder should increase more or less linearly with the nest-feeder distance. Our results unambiguously support the second strategy. When feeders were established at distances of 5-75 m from the nest, the distances steered downwind of the food increased from 0.7 m to 3.4 m in a linear fashion. This result was independent of wind speed or wind direction. It translates into an ant's estimate of its navigation error within a range of 3 degrees to 8 degrees. PMID- 16272246 TI - Year-round recordings of behavioural and physiological parameters reveal the survival strategy of a poorly insulated diving endotherm during the Arctic winter. AB - Warm-blooded diving animals wintering in polar regions are expected to show a high degree of morphological adaptation allowing efficient thermal insulation. In stark contrast to other marine mammals and seabirds living at high latitudes, Arctic great cormorants Phalacrocorax carbo have very limited thermal insulation because of their partly permeable plumage. They nonetheless winter in Greenland, where they are exposed to very low air and water temperatures. To understand how poorly insulated diving endotherms survive the Arctic winter, we performed year round recordings of heart rate, dive depth and abdominal temperature in male great cormorants using miniature data loggers. We also examined the body composition of individuals in the spring. Abdominal temperatures and heart rates of birds resting on land and diving showed substantial variability. However, neither hypothermia nor significantly lower heart rate levels were recorded during the winter months. Thus our data show no indication of general metabolic depression in great cormorants wintering in Greenland. Furthermore, great cormorants did not reduce their daily swimming time during the coldest months of the year to save energy; they continued to forage in sub-zero waters for over an hour every day. As birds spent extended periods in cold water and showed no signs of metabolic depression during the Arctic winter, their theoretical energy requirements were substantial. Using our field data and a published algorithm we estimated the daily food requirement of great cormorants wintering in Greenland to be 1170+/-110 g day(-1). This is twice the estimated food requirement of great cormorants wintering in Europe. Great cormorants survive the Arctic winter but we also show that they come close to starvation during the spring, with body reserves sufficient to fast for less than 3 days. Lack of body fuels was associated with drastically reduced body temperatures and heart rates in April and May. Concurrent, intense feeding activity probably allowed birds to restore body reserves. Our study is the first to record ecophysiological parameters in a polar animal on a year-round basis. It challenges the paradigm that efficient thermal insulation is a prerequisite to the colonization of polar habitats by endotherms. PMID- 16272247 TI - Expression of eight distinct MHC isoforms in bovine striated muscles: evidence for MHC-2B presence only in extraocular muscles. AB - This study aimed to analyse the expression of myosin heavy chain (MHC) isoforms in bovine muscles, with particular attention to the MHC-2B gene. Diaphragm, longissimus dorsi, masseter, several laryngeal muscles and two extraocular muscles (rectus lateralis and retractor bulbi) were sampled in adult male Bos taurus (age 18-24 months, mass 400-500 kg) and analysed by RT-PCR, gel electrophoresis and immunohistochemistry. Transcripts and proteins corresponding to eight MHC isoforms were identified: MHC-alpha and MHC-beta/slow (or MHC-1), two developmental isoforms (MHC-embryonic and MHC-neonatal), three adult fast isoforms (MHC-2A, MHC-2X and MHC-2B) and the extraocular isoform MHC-Eo. All eight MHC isoforms were found to be co-expressed in extrinsic eye muscles, retractor bulbi and rectus lateralis, four (beta/slow, 2A, 2X, neonatal) in laryngeal muscles, three (beta/slow, 2A and 2X) in trunk and limb muscles and two (beta/slow and alpha) in masseter. The expression of MHC-2B and MHC-Eo was restricted to extraocular muscles. Developmental MHC isoforms (neonatal and embryonic) were only found in specialized muscles in the larynx and in the eye. MHC-alpha was only found in extraocular and masseter muscle. Single fibres dissected from masseter, diaphragm and longissimus were classified into five groups (expressing, respectively, beta/slow, alpha, slow and 2A, 2A and 2X) on the basis of MHC isoform electrophoretical separation, and their contractile properties [maximum shortening velocity (v(0)) and isometric tension (P(0))] were determined. v(0) increased progressively from slow to fast 2A and fast 2X, whereas hybrid 1-2A fibres and fibres containing MHC-alpha were intermediate between slow and fast 2A. PMID- 16272248 TI - The red muscle morphology of the thresher sharks (family Alopiidae). AB - A more medial and anterior position of the red aerobic myotomal muscle (RM) and the presence of a vascular counter-current heat exchange system provide the functional elements that facilitate regional RM endothermy in tunas, lamnid sharks and the common thresher shark (Alopias vulpinus). The convergent RM morphology among all species capable of RM endothermy suggests that RM position is a strong predictor of fish endothermic capacity. The present study investigated the comparative RM morphology of the other two thresher shark species (bigeye thresher, Alopias superciliosus, and the pelagic thresher, Alopias pelagicus), for which there is no information regarding their capacity for RM endothermy, and compared these data with published works on A. vulpinus. The digitization of transverse sections along the body of A. superciliosus and A. pelagicus enabled quantification of the relative amount of RM and the position and placement of the RM along the body. The RM in both A. superciliosus and A. pelagicus is positioned subcutaneously, along the lateral edges of the myotomes, and is distributed relatively evenly over the trunk of the body. The position of maximum RM area is at 50% fork length (FL) for A. superciliosus and at 75% FL for A. pelagicus. The amount of RM (mean +/- S.E.M.) is 2.31+/-0.11% and 3.01+/-0.10% in A. superciliosus and A. pelagicus, respectively. When compared with A. vulpinus, all three alopiid sharks have a similar amount of RM. However, A. superciliosus and A. pelagicus differ from A. vulpinus in that they do not possess the medial and anterior RM arrangement that would likely facilitate metabolic heat conservation (RM endothermy). PMID- 16272249 TI - The effects of L-arginine and L-NAME supplementation on redox-regulation and thermogenesis in interscapular brown adipose tissue. AB - Changes in inducible nitric oxide synthase (iNOS) protein levels and its relationship with the hyperplasia and uncoupling protein 1 (UCP1) levels were examined in interscapular brown adipose tissue (IBAT) of adult rat males receiving L-arginine (L-Arg; 2.25%) or N-nitro-L-arginine methyl ester (L-NAME; 0.01%) as a drinking liquid and maintained at low (4+/-1 degrees C) or room (22+/ 1 degrees C) temperature for 45 days. Cold generally diminished both iNOS immunopositivity and protein level in IBAT, as well as the rate of apoptosis. Among groups acclimated to cold, higher iNOS immunopositivity and protein levels were detected only in the L-Arg-treated group. Furthermore, chronic L-Arg treatment increased IBAT mass and UCP1 protein content, while L-NAME had an opposite effect, decreasing both IBAT mass and UCP1 protein level, as compared to the control maintained at 4+/-1 degrees C. These data suggest that nitric oxide (NO) produced by iNOS could also contribute to overall NO-associated regulation of thermogenesis in IBAT. Namely, that iNOS, i.e. NO, in correlation with enhanced thermogenesis, additionally induced IBAT hyperplasia and UCP1 level compared to that induced by low temperature. Cooperative action of decreased apoptosis accompanied by increased tissue hyperplasia and UCP1 level, observed in IBAT of cold-acclimated rats, would be a way of meeting the metabolic requirements for increased thermogenesis. PMID- 16272250 TI - Molecular characterisation of SALMFamide neuropeptides in sea urchins. AB - The SALMFamides are a family of neuropeptides found in species belonging to the phylum Echinodermata. Members of this family have been identified in starfish (class Asteroidea) and in sea cucumbers (class Holothuroidea) but not in other echinoderms. Our aim here was to characterise SALMFamide neuropeptides in sea urchins (class Echinoidea). Radioimmunoassays for the starfish SALMFamides S1 and S2 were used to test for related peptides in whole-body acetone extracts of the sea urchin Echinus esculentus. Fractionation of extracts using high performance liquid chromatography (HPLC) revealed several peaks of SALMFamide-like immunoreactivity, with two S2-like immunoreactive peaks (3 and 4) being the most prominent. However, peak 4 could not be purified to homogeneity and although peak 3 was purified, only a partial sequence (MRYH) could be obtained. An alternative strategy for identification of echinoid SALMFamides was provided by sequencing the genome of the sea urchin Strongylocentrotus purpuratus. Analysis of whole genome shotgun sequence data using the Basic Local Alignment Search Tool (BLAST) identified a contig (347664) that contains a coding region for seven putative SALMFamide neuropeptides (PPVTTRSKFTFamide, DAYSAFSFamide, GMSAFSFamide, AQPSFAFamide, GLMPSFAFamide, PHGGSAFVFamide and GDLAFAFamide), which we have named SpurS1-SpurS7, respectively. Three of these peptides (SpurS1-3) have the C terminal sequences TFamide or SFamide, which are identical or similar to the C terminal region of the starfish SALMFamide S2. This may explain the occurrence of several S2-like immunoreactive peptides in extracts of Echinus esculentus. Detailed analysis of the sequence of contig 347664 indicated that the SALMFamide gene in Strongylocentrotus purpuratus comprises two exons, with the first exon encoding a signal peptide sequence and the second exon encoding SpurS1-SpurS7. Characterisation of this gene is important because it is the first echinoderm neuropeptide precursor sequence to be identified and, more specifically, it provides our first insight into the structure and organisation of a SALMFamide gene in an echinoderm. In particular, it has revealed a hitherto unknown complexity in the diversity of SALMFamide neuropeptides that may occur in an echinoderm species because all previous studies, which relied on peptide purification and sequencing, revealed only two SALMFamide neuropeptides in each species examined. It now remains to be established whether or not the occurrence of more than two SALMFamides in Strongylocentrotus purpuratus is a feature that is peculiar to this species and to echinoids in general or is more widespread across the phylum Echinodermata. Identification of SpurS1-SpurS7 provides the basis for comparative analysis of the physiological actions of these peptides in sea urchins and for exploitation of the sea urchin genome sequence to identify the receptor(s) that mediate effects of SALMFamides in echinoderms. PMID- 16272251 TI - Thermally induced changes in lipid composition of raft and non-raft regions of hepatocyte plasma membranes of rainbow trout. AB - In poikilotherms, increases in plasma membrane (PM) cholesterol and an increase in the degree of lipid acyl chain saturation commonly accompany an increase in growth temperature. This has typically been interpreted in terms of membrane fluidity/order homeostasis, but these changes would also be expected to stabilize the structure of PM rafts against thermal perturbation. Rafts are microdomains that organize the molecules of many signaling cascades and are formed as a result of interactions between lipids with saturated acyl chains and cholesterol. No study to date has examined the thermally induced compositional changes of raft and non-raft regions of the PM separately. In this study we have measured the phospholipid class composition and fatty acid composition of raft-enriched (raft) and raft-depleted PM (RDPM) of hepatocytes from trout Oncorhynchus mykiss acclimated to 5 degrees C and 20 degrees C. In the raft, warm acclimation was associated with a reduction in the proportion of phosphatidylcholine from 56% to 30% while phosphatidylserine and phosphatidylinositol each increased from 8% to approximately 20% of the total phospholipid. Additionally, there were significantly fewer unsaturated fatty acids in the raft lipids from warm acclimated (61%) than from the cold-acclimated trout (68%). In contrast, there were no significant changes in phospholipid class or acyl chain unsaturation in the RDPM. These data suggest that changes in raft lipid composition, rather than the PM as a whole, are particularly important during thermal acclimation. PMID- 16272252 TI - Time course of osmoregulatory and metabolic changes during osmotic acclimation in Sparus auratus. AB - Changes in different osmoregulatory and metabolic parameters over time were assessed in gills, kidney, liver and brain of gilthead sea bream Sparus auratus transferred either from seawater (SW, 38 p.p.t.) to hypersaline water (HSW, 55 p.p.t.) or from SW to low salinity water (LSW, 6 p.p.t.) for 14 days. Changes displayed by osmoregulatory parameters revealed two stages during hyperosmotic and hypo-osmotic acclimation: (i) an adaptive period during the first days of acclimation (1-3 days), with important changes in these parameters, and (ii) a chronic regulatory period (after 3 days of transfer) where osmotic parameters reached homeostasis. From a metabolic point of view, two clear phases can also be distinguished during acclimation to hyperosmotic or hypo-osmotic conditions. The first one coincides with the adaptive period and is characterized by enhanced levels of plasma metabolites (glucose, lactate, triglycerides and protein), and use of these metabolites by different tissues in processes directly or indirectly involved in osmoregulatory work. The second stage coincides with the chronic regulatory period observed for the osmoregulatory parameters and is metabolically characterized in HSW-transferred fish by lower energy expenditure and a readjustment of metabolic parameters to levels returning to normality, indicative of reduced osmoregulatory work in this stage. In LSW-transferred fish, major changes in the second stage include: (i) decreased glycolytic potential, capacity for exporting glucose and potential for amino acid catabolism in liver; (ii) enhanced use of exogenous glucose through glycolysis, pentose phosphate and glycogenesis in gills; (iii) increased glycolytic potential in kidney; and (iv) increased glycogenolytic potential and capacity for use of exogenous glucose in brain. PMID- 16272254 TI - Neural plasticity of mushroom body-extrinsic neurons in the honeybee brain. AB - Central interneurons exiting the alpha lobe of the mushroom bodies were studied with respect to their plasticity by electrically stimulating their presynaptic inputs, the Kenyon cells. Special attention was given to the analysis of a single, identified neuron, the PE1. Three stimulation protocols were tested: double pulses, tetanus (100 Hz for 1 s), and tetanus paired with intracellular de or hyper-polarization of the recorded cell. Double-pulse stimulations revealed short-term facilitation and depression, tuning the responses of these interneurons to frequencies in the range of 20-40 Hz. The tetanus may lead to augmentation of responses to test stimuli lasting for several minutes, or to depression followed by augmentation. Associative long-term potentiation (LTP) was induced in the PE1 neuron by pairing a presynaptic tetanus with depolarization. This is the first time that associative LTP has been found in an interneuron of the insect nervous system. These data are discussed in the context of spike tuning in the output of the mushroom body, and the potential role of associative LTP in olfactory learning. It is concluded that the honeybee mushroom body output neurons are likely to contribute to the formation of olfactory memory. PMID- 16272253 TI - Transepithelial urate transport by avian renal proximal tubule epithelium in primary culture. AB - Birds are uricotelic, and because they excrete urate by renal tubular secretion, they provide a convenient model for examination of this process. Primary monolayer cultures of the isolated renal proximal tubule epithelium from the domestic chicken, Gallus gallus L., were mounted in Ussing chambers where several substrates/inhibitors of renal organic anion transporters were tested for the sidedness and specificity of their effects on transepithelial urate transport. Transepithelial electrical resistance, electrical potential and sodium-dependent glucose current were monitored to detect nonspecific effects. Under control short circuited conditions the ratio of unidirectional fluxes of [(14)C]urate was found to be 3:1. Active net secretion was specifically inhibited by 1 mmol l(-1) probenecid and 10 mmol l(-1) para-aminohippuric acid (PAH). Bromocresol Green, cimetidine, nocodozole, cytochalasin D and ouabain also inhibited secretion but were toxic. Interstitial-side lithium (5 mmol l(-1)) and glutarate (1 mmol l(-1)) specifically blocked transport, but 10-100 micromol l(-1) glutarate had no effect. Interstitial estrone sulfate (ES) stimulated urate secretion at 10 micromol l(-1) but was inhibitory at 500 micromol l(-1). Active PAH secretion (5:1 flux ratio) was inhibited 34% by 330 micromol l(-1) urate. ES (500 micromol l(-1)) blocked the remainder. From the lumen side, glucose-free, Cl(-)-free and high K(+) (30 mmol l(-1)) solutions, or an alkaline pH of 7.7 had no effect on urate transport and neither did several compounds known to be uricosuric. Lumen side methotrexate (500 micromol l(-1)) and MK571 (20 micromol l(-1)) strongly inhibited urate secretion. MK571 had no effect from the interstitial side. RT-PCR revealed mRNA for OAT1-, OAT3-, MRP2- and MRP4-like organic anion transporters in chicken proximal epithelium. PMID- 16272255 TI - Respiratory and acid-base responses during migration and to exercise by the terrestrial crab Discoplax (Cardisoma) hirtipes, with regard to season, humidity and behaviour. AB - The terrestrial crab Discoplax hirtipes (formerly Cardisoma hirtipes) exhibits a seasonally dichotomous activity pattern governed by the seasonal rainfall on Christmas Island, with a breeding migration in the wet season. Greater activity in the wet season reflects a release of constraints on ion and water balance independent of changes in exercise physiology. The respiratory responses to walking exercise by D. hirtipes were assessed with regard to humidity and season. In the laboratory, crabs walked 38% faster when humidity was high, while the O(2) uptake across the lung became diffusion limited despite a Ca(2+)-induced increase in O(2) affinity of the haemocyanin. Crabs walking in 90% relative humidity exhibited a larger metabolic acidosis while re-oxidation of l-lactate was only 0.49 mmol l(-1) h(-1). The wet-season crabs were more active and exhibited a respiratory acidosis compared with the quiescent dry-season crabs. The migration was close to the limit of the aerobic scope of the crabs, and the pulmonary oxygen partial pressure (P(O(2))) was depressed, the O(2) uptake diffusion limited and the arterial-venous P(O(2)) difference diminished. Additional, enforced exercise induced a metabolic acidosis. The optimum strategy for migration would be walking continuously but at a speed within the aerobic scope. D. hirtipes is influenced by seasonal rain and responds to lower ambient humidity by limiting exercise. The behavioural response is paramount since the changes in respiratory status were determined primarily by the differences in commitment to, and investment in, walking and not by direct effects of humidity on respiratory physiology. PMID- 16272256 TI - Actin cytoskeleton of rabbit intestinal cells is a target for potent marine phycotoxins. AB - Biotoxins produced by harmful marine microalgae (phycotoxins) can be accumulated into seafood, representing a great risk for public health. Some of these phycotoxins are responsible for a variety of gastrointestinal disturbances; however, the relationship between their mechanism of action and toxicity in intestinal cells is still unknown. The actin cytoskeleton is an important and highly complicated structure in intestinal cells, and on that basis our aim has been to investigate the effect of representative phycotoxins on the enterocyte cytoskeleton. We have quantified for the first time the loss of enterocyte microfilament network induced by each toxin and recorded fluorescence images using a laser-scanning cytometer and confocal microscopy. Our data show that pectenotoxin-6, maitotoxin, palytoxin and ostreocin-D cause a significant reduction in the actin cytoskeleton. In addition, we found that the potency of maitotoxin, palytoxin and ostreocin-D to damage filamentous actin is related to Ca(2+) influx in enterocytes. Those results identify the cytoskeleton as an early target for the toxic effect of those toxins. PMID- 16272258 TI - Non-hotspot-related breakpoints of common deletions in Sotos syndrome are located within destabilised DNA regions. AB - BACKGROUND: Sotos syndrome (SoS) is a disorder characterised by excessive growth, typical craniofacial features, and developmental retardation. It is caused by haploinsuffiency of NSD1 at 5q35. There is a 3.0 kb recombination hotspot in which the breakpoints of around 80% of SoS patients with a common deletion can be mapped. OBJECTIVE: To identify deletion breakpoints located outside the SoS recombination hotspot. METHODS: A screening system for the directly orientated segments of the SoS LCRs was developed for 10 SoS patients with a common deletion who were negative for the SoS hotspot. Deletion-junction fragments were analysed for DNA duplex stability and their relation to scaffold/matrix attachment regions (S/MARs). These features were compared with the SoS hotspot and recombination hotspots of other genomic disorders. RESULTS: The breakpoint was mapped in four SoS patients, two with a deletion in the maternally derived chromosome. These breakpoint regions were located approximately 2.5 kb, approximately 9.6 kb, approximately 27.2, and approximately 27.7 kb telomeric to the SoS hotspot and were confined to 164 bp, 46 bp, 256 bp, and 124 bp, respectively. Two of the regions were mapped within Alu elements. All crossover events were found to have occurred within or adjacent to a highly destabilised DNA duplex with a high S/MAR probability. In contrast, the SoS hotspot and other genomic disorders' recombination hotspots were mapped to stabilised DNA helix regions, flanked by destabilised regions with high probability of containing S/MAR elements. CONCLUSIONS: The data suggest that a specific chromatin structure may increase susceptibility for recurrent crossover events and thus predispose to recombination hotspots in genomic disorders. PMID- 16272257 TI - The G6055A (G2019S) mutation in LRRK2 is frequent in both early and late onset Parkinson's disease and originates from a common ancestor. AB - BACKGROUND: Mutations in the gene Leucine-Rich Repeat Kinase 2 (LRRK2) were recently identified as the cause of PARK8 linked autosomal dominant Parkinson's disease. OBJECTIVE: To study recurrent LRRK2 mutations in a large sample of patients from Italy, including early (<50 years) and late onset familial and sporadic Parkinson's disease. RESULTS: Among 629 probands, 13 (2.1%) were heterozygous carriers of the G2019S mutation. The mutation frequency was higher among familial (5.1%, 9/177) than among sporadic probands (0.9%, 4/452) (p<0.002), and highest among probands with one affected parent (8.7%, 6/69) (p<0.001). There was no difference in the frequency of the G2019S mutation in probands with early v late onset disease. Among 600 probands, one heterozygous R1441C but no R1441G or Y1699C mutations were detected. None of the four mutations was found in Italian controls. Haplotype analysis in families from five countries suggested that the G2019S mutation originated from a single ancient founder. The G2019S mutation was associated with the classical Parkinson's disease phenotype and a broad range of onset age (34 to 73 years). CONCLUSIONS: G2019S is the most common genetic determinant of Parkinson's disease identified so far. It is especially frequent among cases with familial Parkinson's disease of both early and late onset, but less common among sporadic cases. These findings have important implications for diagnosis and genetic counselling in Parkinson's disease. PMID- 16272259 TI - Identification of mutations in the AIPL1, CRB1, GUCY2D, RPE65, and RPGRIP1 genes in patients with juvenile retinitis pigmentosa. AB - OBJECTIVE: To identify mutations in the AIPL1, CRB1, GUCY2D, RPE65, and RPGRIP1 genes in patients with juvenile retinitis pigmentosa. METHODS: Mutation analysis was carried out in a group of 35 unrelated patients with juvenile autosomal recessive retinitis pigmentosa (ARRP), Leber's congenital amaurosis (LCA), or juvenile isolated retinitis pigmentosa (IRP), by denaturing high performance liquid chromatography followed by direct sequencing. RESULTS: All three groups of patients showed typical combinations of eye signs associated with retinitis pigmentosa: pale optic discs, narrow arterioles, pigmentary changes, and nystagmus. Mutations were found in 34% of PATIENTS: in CRB1 (11%), GUCY2D (11%), RPE65 (6%), and RPGRIP1 (6%). Nine mutations are reported, including a new combination of two mutations in CRB1, and new mutations in GUCY2D and RPGRIP1. The new GUCY2D mutation (c.3283delC, p.Pro1069ArgfsX37) is the first pathological sequence change reported in the intracellular C-terminal domain of GUCY2D, and did not lead to the commonly associated LCA, but to a juvenile retinitis pigmentosa phenotype. The polymorphic nature of three previously described (pathological) sequence changes in AIPL1, CRB1, and RPGRIP1 was established. Seven new polymorphic changes, useful for further association studies, were found. CONCLUSIONS: New and previously described sequence changes were detected in retinitis pigmentosa in CRB1, GUCY2D, and RPGRIP1; and in LCA patients in CRB1, GUCY2D, and RPE65. These data, combined with previous reports, suggest that LCA and juvenile ARRP are closely related and belong to a continuous spectrum of juvenile retinitis pigmentosa. PMID- 16272260 TI - Genetics of skin appendage neoplasms and related syndromes. AB - In the past decade the molecular basis of many inherited syndromes has been unravelled. This article reviews the clinical and genetic aspects of inherited syndromes that are characterised by skin appendage neoplasms, including Cowden syndrome, Birt-Hogg-Dube syndrome, naevoid basal cell carcinoma syndrome, generalised basaloid follicular hamartoma syndrome, Bazex syndrome, Brooke Spiegler syndrome, familial cylindromatosis, multiple familial trichoepitheliomas, and Muir-Torre syndrome. PMID- 16272261 TI - Further evidence for LBP-1c/CP2/LSF association in Alzheimer's disease families. AB - OBJECTIVES: Several studies suggested chromosome 12 harbours an Alzheimer's disease (AD) risk factor gene. Significant association of a single nucleotide polymorphism (SNP) in the 3' UTR of transcription factor CP2 (LBP-1c/CP2/LSF or TFCP2) at 12q13 was reported in three independent case-control studies, but no family based analyses have been performed to date. METHODS: Genotypes for three SNPs were generated in two independent AD family samples. A meta-analysis on all published case-control studies was also performed. RESULTS: The A allele of the 3' UTR SNP was associated with increased risk for AD in one sample (odds ratio (OR) 2.1, 95% confidence interval (95% CI) 1.1 to 4.3), but not in the other, possibly due to low power. Haplotype analyses showed that this allele is part of a putative risk-haplotype overtransmitted to affected individuals in one sample and in both samples combined. Meta-analysis of the previously associated 3' UTR SNP showed a trend towards a protective effect of the A allele in AD (OR 0.73, 95% CI 0.5 to 1.1). CONCLUSIONS: This is the first study to examine LBP 1c/CP2/LSF in AD families, and the fifth to independently show significant association. While our results support a role of this gene in AD pathogenesis, the direction of the effect remains uncertain, possibly indicating linkage disequilibrium with another variant nearby. PMID- 16272262 TI - Catecholaminergic polymorphic ventricular tachycardia: RYR2 mutations, bradycardia, and follow up of the patients. AB - BACKGROUND: The aim of the study was to assess underlying genetic cause(s), clinical features, and response to therapy in catecholaminergic polymorphic ventricular tachycardia (CPVT) probands. METHODS AND RESULTS: We identified 13 missense mutations in the cardiac ryanodine receptor (RYR2) in 12 probands with CPVT. Twelve were new, of which two are de novo mutations. A further 11 patients were silent gene carriers, suggesting that some mutations are associated with low penetrance. A marked resting sinus bradycardia off drugs was observed in all carriers. On beta blocker treatment, 98% of the RYR2 mutation carriers remained symptom free with a median follow up of 2 (range: 2-37) years. CONCLUSION: CPVT patients with RYR2 mutation have bradycardia regardless of the site of the mutation, which could direct molecular diagnosis in (young) patients without structural heart disease presenting with syncopal events and a slow heart rate but with normal QTc at resting ECG. Treatment with beta blockers has been very effective in our CPVT patients during initial or short term follow up. Given the risk of sudden death and the efficacy of beta blocker therapy, the identification of large numbers of RYR2 mutations thus calls for genetic screening, early diagnosis, and subsequent preventive strategies. PMID- 16272263 TI - The otic ganglion in rats and its parotid connection: cholinergic pathways, reflex secretion and a secretory role for the facial nerve. AB - Otic ganglionectomy in rats was found to have affected the parotid gland more profoundly than section of the auriculotemporal nerve as assessed by reduction in gland weight (by 33 versus 20%) and total acetylcholine synthesizing capacity (by 88 versus 76%) 1 week postoperatively and, when assessed on the day of surgery under adrenoceptor blockade, by reflex secretion (by 99 versus 88%). The facial nerve contributed to the acetylcholine synthesizing capacity of the gland. Section of the nerve only, at the level of the stylomastoid foramen, reduced the acetylcholine synthesis by 15% and, combined with otic ganglionectomy, by 98% or, combined with section of the auriculotemporal nerve, by 82%. The facial nerve was secretory to the gland, and the response was of a cholinergic nature. The nerve conveyed reflex secretion of saliva and caused secretion of saliva upon stimulation. Most of the facial secretory nerve fibres originated from the otic ganglion, since after otic ganglionectomy (and allowing for nerve degeneration) the secretory response to facial nerve stimulation was markedly reduced (from 23 to 4 microl (5 min)(-1)). The persisting secretory response after otic ganglionectomy, exaggerated due to sensitization, and the residual acetylcholine synthesizing capacity (mainly depending on the facial nerve) showed that a minor proportion of pre- and postganglionic nerve fibres relay outside the otic ganglion. The great auricular nerve, which like the facial nerve penetrates the gland, caused no secretion of saliva upon stimulation. Avulsion of the auriculotemporal nerve was more effective than otic ganglionectomy in reducing the acetylcholine synthesizing capacity (by 94 versus 88%) and as effective as otic ganglionectomy in abolishing reflex secretion (by 99%). When aiming at parasympathetic denervation, avulsion may be the preferable choice, since it is technically easier to perform than otic ganglionectomy. PMID- 16272264 TI - Breath-holding and its breakpoint. AB - This article reviews the basic properties of breath-holding in humans and the possible causes of the breath at breakpoint. The simplest objective measure of breath-holding is its duration, but even this is highly variable. Breath-holding is a voluntary act, but normal subjects appear unable to breath-hold to unconsciousness. A powerful involuntary mechanism normally overrides voluntary breath-holding and causes the breath that defines the breakpoint. The occurrence of the breakpoint breath does not appear to be caused solely by a mechanism involving lung or chest shrinkage, partial pressures of blood gases or the carotid arterial chemoreceptors. This is despite the well-known properties of breath-hold duration being prolonged by large lung inflations, hyperoxia and hypocapnia and being shortened by the converse manoeuvres and by increased metabolic rate. Breath-holding has, however, two much less well-known but important properties. First, the central respiratory rhythm appears to continue throughout breath-holding. Humans cannot therefore stop their central respiratory rhythm voluntarily. Instead, they merely suppress expression of their central respiratory rhythm and voluntarily 'hold' the chest at a chosen volume, possibly assisted by some tonic diaphragm activity. Second, breath-hold duration is prolonged by bilateral paralysis of the phrenic or vagus nerves. Possibly the contribution to the breakpoint from stimulation of diaphragm muscle chemoreceptors is greater than has previously been considered. At present there is no simple explanation for the breakpoint that encompasses all these properties. PMID- 16272265 TI - Muscle glycogen reduction in man: relationship between surface EMG activity and oxygen uptake kinetics during heavy exercise. AB - The purpose of this study was to determine whether muscle glycogen reduction prior to exercise would alter muscle fibre recruitment pattern and change either on-transient O2 uptake (VO2) kinetics or the VO2 slow component. Eight recreational cyclists (VO2peak, 55.6 +/- 1.3 ml kg (-1) min(-1)) were studied during 8 min of heavy constant-load cycling performed under control conditions (CON) and under conditions of reduced type I muscle glycogen content (GR). VO2 was measured breath-by-breath for the determination of VO2 kinetics using a double-exponential model with independent time delays. VO2 was higher in the GR trial compared to the CON trial as a result of augmented phase I and II amplitudes, with no difference between trials in the phase II time constant or the magnitude of the slow component. The mean power frequency (MPF) of electromyography activity for the vastus medialis increased over time during both trials, with a greater rate of increase observed in the GR trial compared to the CON trial. The results suggest that the recruitment of additional type II motor units contributed to the slow component in both trials. An increase in fat metabolism and augmented type II motor unit recruitment contributed to the higher VO2 in the GR trial. However, the greater rate of increase in the recruitment of type II motor units in the GR trial may not have been of sufficient magnitude to further elevate the slow component when VO2 was already high and approaching VO2peak . PMID- 16272268 TI - Acute compartment syndrome in lower extremity musculoskeletal trauma. AB - Acute compartment syndrome is a potentially devastating condition in which the pressure within an osseofascial compartment rises to a level that decreases the perfusion gradient across tissue capillary beds, leading to cellular anoxia, muscle ischemia, and death. A variety of injuries and medical conditions may initiate acute compartment syndrome, including fractures, contusions, bleeding disorders, burns, trauma, postischemic swelling, and gunshot wounds. Diagnosis is primarily clinical, supplemented by compartment pressure measurements. Certain anesthetic techniques, such as nerve blocks and other forms of regional and epidural anesthesia, reportedly contribute to a delay in diagnosis. Basic science data suggest that the ischemic threshold of normal muscle is reached when pressure within the compartment is elevated to 20 mm Hg below the diastolic pressure or 30 mm Hg below the mean arterial blood pressure. On diagnosis of impending or true compartment syndrome, immediate measures must be taken. Complete fasciotomy of all compartments involved is required to reliably normalize compartment pressures and restore perfusion to the affected tissues. Recognizing compartment syndromes requires having and maintaining a high index of suspicion, performing serial examinations in patients at risk, and carefully documenting changes over time. PMID- 16272269 TI - Direct catastrophic injury in sports. AB - Catastrophic sports injuries are rare but tragic events. Direct (traumatic) catastrophic injury results from participating in the skills of a sport, such as a collision in football. Football is associated with the greatest number of direct catastrophic injuries for all major team sports in the United States. Pole vaulting, gymnastics, ice hockey, and football have the highest incidence of direct catastrophic injuries for sports in which males participate. In most sports, the rate of catastrophic injury is higher at the collegiate than at the high school level. Cheerleading is associated with the highest number of direct catastrophic injuries for all sports in which females participate. Indirect (nontraumatic) injury is caused by systemic failure as a result of exertion while participating in a sport. Cardiovascular conditions, heat illness, exertional hyponatremia, and dehydration can cause indirect catastrophic injury. Understanding the common mechanisms of injury and prevention strategies for direct catastrophic injuries is critical in caring for athletes. PMID- 16272270 TI - Preoperative planning for primary total hip arthroplasty. AB - Preoperative planning is of paramount importance in obtaining reproducible results in modern hip arthroplasty. Planning helps the surgeon visualize the operation after careful review of the clinical and radiographic findings. A standardized radiograph with a known magnification should be used for templating. The cup template should be placed relative to the ilioischial line, the teardrop, and the superolateral acetabular margin, so that the removal of the supportive subchondral bone is minimal and the center of rotation of the hip is restored. When acetabular abnormalities are encountered, additional measures are necessary to optimize cup coverage and minimize the risk of malposition. Templating the femoral side for cemented and cementless implants should aim to optimize limb length and femoral offset, thereby improving the biomechanics of the hip joint. Meticulous preoperative planning allows the surgeon to perform the procedure expediently and precisely, anticipate potential intraoperative complications, and achieve reproducible results. PMID- 16272271 TI - Rheumatoid arthritis in the cervical spine. AB - The cervical spine often becomes involved early in the course of rheumatoid arthritis, leading to three different patterns of instability: atlantoaxial subluxation, atlantoaxial impaction, and subaxial subluxation. Although radiographic changes are common, the prevalence of neurologic injury is relatively low. The primary goal of treatment is to prevent permanent neurologic injury while avoiding potentially dangerous and unnecessary surgery. Strategies include patient education, lifestyle modification, regular radiographic follow up, and early surgical intervention, when indicated. Magnetic resonance imaging is indicated when neurologic deficit (myelopathy) occurs or when plain radiographs show atlantoaxial subluxation with a posterior atlantodental interval < or =14 mm, any degree of atlantoaxial impaction, or subaxial stenosis with a canal diameter < or =14 mm. Surgery should be considered promptly for any of the following: progressive neurologic deficit, chronic neck pain in the setting of radiographic instability that does not respond to nonnarcotic pain medication, any degree of atlantoaxial impaction or cord stenosis, a posterior atlantodental interval < or =14 mm, atlantoaxial impaction represented by odontoid migration > or =5 mm rostral to McGregor's line, sagittal canal diameter <14 mm, or a cervicomedullary angle <135 degrees. PMID- 16272272 TI - Secondary osteoporosis. AB - Secondary osteoporosis occurs as a consequence of various lifestyle factors (eg, eating disorders, smoking, alcoholism), disease processes (eg, endocrinopathies, gastrointestinal tract disease, hepatobiliary disease), and treatment regimens that comprise corticosteroids or chemotherapeutic agents. Some of the disease entities underlying secondary osteoporosis may be clinically silent and identified only during evaluation for documented osteoporosis. The pathogenesis of osteoporosis in these settings is typically multifactorial. The loss of bone may be direct or indirect but ultimately is related to altered osteoblast or osteoclast function. Causes of secondary osteoporosis should especially be investigated in men at all ages and in premenopausal women with atraumatic fractures. In addition, patients with known risk factors should be evaluated. Early recognition and intervention are essential to prevent further loss of bone mass and to prevent fragility fractures. PMID- 16272273 TI - Arthroscopic Bankart repair. PMID- 16272274 TI - The role of heparan sulfate and TLR2 in cytokine induction by hepatitis B virus capsids. PMID- 16272275 TI - Hypoxia regulates macrophage functions in inflammation. AB - The presence of areas of hypoxia is a prominent feature of various inflamed, diseased tissues, including malignant tumors, atherosclerotic plaques, myocardial infarcts, the synovia of joints with rheumatoid arthritis, healing wounds, and sites of bacterial infection. These areas form when the blood supply is occluded and/or unable to keep pace with the growth and/or infiltration of inflammatory cells in a given area. Macrophages are present in all tissues of the body where they normally assist in guarding against invading pathogens and regulate normal cell turnover and tissue remodeling. However, they are also known to accumulate in large numbers in such ischemic/hypoxic sites. Recent studies show that macrophages then respond rapidly to the hypoxia present by altering their expression of a wide array of genes. In the present study, we outline and compare the phenotypic responses of macrophages to hypoxia in different diseased states and the implications of these for their progression and treatment. PMID- 16272276 TI - Murine vascular endothelium activates and induces the generation of allogeneic CD4+25+Foxp3+ regulatory T cells. AB - Unlike graft-resident donor-derived hemopoietic APCs, which decrease in number over time after transplantation, vascular endothelial cells are lifelong residents of a vascularized allograft. Endothelial cells are potent APCs for allogeneic CD8+ T lymphocytes but are unable to induce proliferation of allogeneic CD4+ T lymphocytes. Although the reason for this differential response has been poorly understood, here we report that alloantigen presentation by vascular endothelium to CD4+ T lymphocytes activates and induces CD4+25+Foxp3+ regulatory T cells, which can inhibit proliferation of alloreactive T cells both in vitro and in vivo. This process occurs independently of B7.1 costimulation but is dependent on programmed death ligand 1 (B7-H1). This finding may have important implications for tolerance induction in transplantation. PMID- 16272277 TI - Yin-yang: balancing act of prostaglandins with opposing functions to regulate inflammation. AB - For many years, cyclooxygenase-2 (COX-2), a critical enzyme for PG production, has been the favorite target for anti-inflammatory drug development. However, recent revelations regarding the adverse effects of selective COX-2 inhibitors have stimulated intense debate. Interestingly, in the early phase of inflammation, COX-2 facilitates inflammatory PG production while in the late phase it has anti-inflammatory effects. Moreover, although some PGs are proinflammatory, others have anti-inflammatory effects. Thus, it is likely that PGs with opposing effects maintain homeostasis, although the molecular mechanism(s) remains unclear. We report here that an inflammatory PG, PGD2, via its receptor, mediates the activation of NF-kappaB stimulating COX-2 gene expression. Most interestingly, an anti-inflammatory PG (PGA1) suppresses NF kappaB activation and inhibits COX-2 gene expression. We propose that while pro- and anti-inflammatory PGs counteract each other to maintain homeostasis, selective COX-2 inhibitors may disrupt this balance, thereby resulting in reported adverse effects. PMID- 16272278 TI - Reciprocal immunomodulation in a schistosome and hepatotropic virus coinfection model. AB - Human coinfection with the helminth parasite Schistosoma mansoni and hepatitis B and hepatitis C viruses is associated with increased hepatic viral burdens and severe liver pathology. In this study we developed a murine S. mansoni/lymphocytic choriomeningitis virus (LCMV) coinfection model that reproduces the enhanced viral replication and liver pathology observed in human coinfections, and used this model to explore the mechanisms involved. Viral coinfection during the Th2-dominated granulomatous phase of the schistosome infection resulted in induction of a strong LCMV-specific T cell response, with infiltration of high numbers of LCMV-specific IFN-gamma-producing CD8+ cells into the liver. This was associated with suppression of production of the Th2 cytokines dominant during S. mansoni infection and a rapid increase in morbidity, linked to hepatotoxicity. Interestingly, the liver of coinfected mice was extremely susceptible to viral replication. This correlated with a reduced intrahepatic type I IFN response following virus infection. Schistosome egg Ags were found to suppress the type I IFN response induced in murine bone marrow derived dendritic cells by polyinosinic-polycytidylic acid. These results suggest that suppression of the antiviral type I IFN response by schistosome egg Ags in vivo predisposes the liver to enhanced viral replication with ensuing immunopathological consequences, findings that may be paralleled in human schistosome/hepatotropic virus coinfections. PMID- 16272279 TI - A selective role for the TNF p55 receptor in autocrine signaling following IFN gamma stimulation in experimental autoimmune uveoretinitis. AB - IFN-gamma stimulates macrophage activation and NO production, which leads to destruction of the retina in experimental autoimmune uveoretinitis. In this study, we investigate the mechanism of disease resistance in TNF p55 receptor deficient animals. We show that although T cell priming is relatively unaffected, macrophages lacking the TNF p55 receptor fail to produce NO following IFN-gamma stimulation because of a requirement for autocrine TNF-alpha signaling through the TNF p55 receptor. In contrast to the impaired activation of NO synthesis, MHC class II up-regulation was indistinguishable in wild-type and TNFRp55-/- mice stimulated with IFN-gamma. These defects could be overcome by stimulating macrophages with LPS. Together, these results show that selected aspects of IFN gamma activation are controlled by autocrine secretion of TNF-alpha, but that this control is lost in the presence of signals generated by pathogen-associated molecular patterns recognizing receptors. PMID- 16272280 TI - Further characterization of reproductive abnormalities in mCd59b knockout mice: a potential new function of mCd59 in male reproduction. AB - CD59 is a GPI-linked membrane protein that inhibits formation of the membrane attack complex of complement. We reported recently that mice have two CD59 genes (termed mCd59a and mCd59b), and that the targeted deletion of mCd59b (mCd59b-/-) results in spontaneous hemolytic anemia and progressive loss of male fertility. Further studies of the reproductive abnormalities in mCd59b-/- mice reported in this study revealed the presence of abnormal multinucleated cells and increased apoptotic cells within the walls of the seminiferous tubules, and a decrease in the number, motility, and viability of sperm associated with a significant increase in abnormal sperm morphologies. Both the capacitation-associated tyrosine phosphorylation and the ionophore-induced acrosome reaction as well as luteinizing hormone, follicle-stimulating hormone, and testosterone serum levels were similar in mCd59b-/- and mCd59b+/+. Surprisingly, the functional deficiency of the complement protein C3 did not rescue the abnormal reproductive phenotype of mCd59b-/-, although it was efficient in rescuing their hemolytic anemia. These results indicate that the male reproductive abnormalities in mCd59b-/- are complement-independent, and that mCd59 may have a novel function in spermatogenesis that is most likely unrelated to its function as an inhibitor of membrane attack complex formation. PMID- 16272281 TI - Essential role of the MyD88 pathway, but nonessential roles of TLRs 2, 4, and 9, in the adjuvant effect promoting Th1-mediated autoimmunity. AB - Induction of tissue-specific experimental autoimmune diseases involves an obligatory adjuvant effect to trigger an innate response of a type that will drive a Th1-biased adaptive response. This is achieved by use of CFA containing mycobacteria (Mycobacterium tuberculosis), whose recognition by cells of the innate immune system depends on TLRs that signal through the adaptor molecule MyD88. We examined the role of selected components of the MyD88 pathway in promoting experimental autoimmune uveitis (EAU). Mice deficient in MyD88, TLR2, TLR4, or TLR9 were immunized with the retinal Ag interphotoreceptor retinoid binding protein in CFA, and their EAU scores and associated immunological responses were examined. MyD88-/- mice were completely resistant to EAU and had a profound defect in Th1, but not Th2, responses to autoantigen challenge. Surprisingly, TLR2-/-, TLR4-/-, and TLR9-/- mice were fully susceptible to EAU and had unaltered adaptive responses to interphotoreceptor retinoid-binding protein. Examination of IL-1R family members, which share the common adaptor MyD88 with the TLR family, revealed that IL-1R-deficient mice, but not IL-18 deficient mice, are resistant to EAU and have profoundly reduced Th1 and Th2 responses. These data are compatible with the interpretation that TLR9, TLR4, and TLR2 signaling is either not needed, or, more likely, redundant in the adjuvant effect needed to induce EAU. In contrast, signaling through the IL-1R plays a necessary and nonredundant role in EAU and can by itself account for the lack of EAU development in MyD88 mice. PMID- 16272282 TI - Permanent survival of fully MHC-mismatched islet allografts by targeting a single chemokine receptor pathway. AB - Chemokine receptor blockade can diminish the recruitment of host effector cells and prolong allograft survival, but little is known of the role of chemokine receptors in promoting host sensitization. We engrafted fully allogeneic islets into streptozotocin-treated normal mice or mice with the autosomal recessive paucity of lymph node T cell (plt) mutation; the latter lack secondary lymphoid expression of the CCR7 ligands, secondary lymphoid organ chemokine (CCL21) and EBV-induced molecule-1 ligand chemokine (CCL19). plt mice showed permanent survival of islets engrafted under the kidney capsule, whereas controls rejected islet allografts in 12 days (p < 0.001), and consistent with this, plt mice had normal allogeneic T cell responses, but deficient migration of donor dendritic cell to draining lymph nodes. Peritransplant i.v. injection of donor splenocytes caused plt recipients to reject their allografts by 12 days, and sensitization at 60 days posttransplant of plt mice with well-functioning allografts restored acute rejection. Finally, islet allografts transplanted intrahepatically in plt mice were rejected approximately 12 days posttransplant, like controls, as were primarily revascularized cardiac allografts. These data show that the chemokine directed homing of donor dendritic cell to secondary lymphoid tissues is essential for host sensitization and allograft rejection. Interruption of such homing can prevent T cell priming and islet allograft rejection despite normal T and B cell functions of the recipient, with potential clinical implications. PMID- 16272283 TI - Immune complex-dependent remodeling of the airway vasculature in response to a chronic bacterial infection. AB - Chronic inflammation in the airways is associated with dramatic architectural changes in the walls of the airways and in the vasculature they contain. In this study, we show that the adaptive immune system is essential for airway remodeling that occurs in mice that are chronically infected with the respiratory pathogen Mycoplasma pulmonis. Angiogenesis, lymphangiogenesis, and epithelial remodeling were greatly reduced in mice that lacked B cells. Substantiating a role for Ab and airway immune complexes, we found that the transfer of immune serum to B cell deficient mice could reconstitute pathogen-induced angiogenesis. Inflammatory cells recruited to the infected airways were activated by the humoral response, and this activation correlated with the induction of genes for remodeling factors such as vascular endothelial growth factor-D. The results reveal a novel pathway whereby T cell-dependent humoral immunity to a persistent airway infection can induce inflammation-dependent angiogenesis, lymphangiogenesis, and chronic airway pathology. PMID- 16272284 TI - Critical requirement of CD11b (Mac-1) on T cells and accessory cells for development of experimental autoimmune encephalomyelitis. AB - Mac-1 (CD18/CD11b) is a member of the beta2-integrin family of adhesion molecules and is implicated in the development of many inflammatory diseases. The role of Mac-1 in the development of CNS demyelinating diseases, including multiple sclerosis, is not understood, and Ab inhibition studies in experimental allergic encephalomyelitis (EAE), the animal model for multiple sclerosis, have produced conflicting findings. To clarify these results and to determine Mac-1-mediated mechanisms in EAE, we performed EAE using Mac-1-deficient mice. Mac-1 homozygous deficient, but not Mac-1 heterozygous-deficient mice, had significantly delayed onset and attenuated EAE. Leukocyte infiltration was similar in both groups of mice in early disease but significantly reduced in spinal cords of receptor deficient mice in late disease. Adoptive transfer of Ag-restimulated T cells from wild-type to Mac-1-deficient mice produced significantly attenuated EAE, whereas transfer of Mac-1-deficient Ag-restimulated T cells to control mice failed to induce EAE. T cells from myelin oligodendrocyte glycoprotein (MOG)35-55 peptide primed Mac-1-deficient mice displayed an altered cytokine phenotype with elevated levels of TGF-beta and IL-10, but reduced levels of IL-2, IFN-gamma, TNF-alpha, IL-12, and IL-4 compared with control mice. Mac-1-deficient T cells from primed mice proliferated comparably to that of control T cells on MOG35-55 restimulation in vitro. However, the draining lymph nodes of MAC-1-deficient mice on day 10 after MOG35-55 immunization contained lower frequency of blast T cells than in control mice, suggesting poor priming. Our results indicate that Mac-1 expression is critical on both phagocytic cells and T cells for the development of demyelinating disease. PMID- 16272285 TI - Ultrasensitive detection and phenotyping of CD4+ T cells with optimized HLA class II tetramer staining. AB - HLA class I tetramers have revolutionized the study of Ag-specific CD8+ T cell responses. Technical problems and the rarity of Ag-specific CD4+ Th cells have not allowed the potential of HLA class II tetramers to be fully realized. Here, we optimize HLA class II tetramer staining methods through the use of a comprehensive panel of HIV-, influenza-, CMV-, and tetanus toxoid-specific tetramers. We find rapid and efficient staining of DR1- and DR4-restricted CD4+ cell lines and clones and show that TCR internalization is not a requirement for immunological staining. We combine tetramer staining with magnetic bead enrichment to detect rare Ag-specific CD4+ T cells with frequencies as low as 1 in 250,000 (0.0004% of CD4+ cells) in human PBLs analyzed directly ex vivo. This ultrasensitive detection allowed phenotypic analysis of rare CD4+ T lymphocytes that had experienced diverse exposure to Ag during the course of viral infections. These cells would not be detectable with normal flow-cytometric techniques. PMID- 16272286 TI - CD1a-, b-, and c-restricted TCRs recognize both self and foreign antigens. AB - Individual CD1-restricted T cells can recognize either endogenous or foreign lipid Ags, but the extent to which the same CD1-restricted TCR can react to both self and microbial lipids is unknown. In this study, we have identified CD1a-, CD1b-, and CD1c-restricted T cells from normal human donors that induce cytolysis and secrete copious IFN-gamma in response to self-CD1 expressed on monocyte derived dendritic cells. Remarkably, microbial Ags presented by CD1 are even more potent agonists for these same T cells. The alphabeta T cell receptors from such clones are diverse and confer specificity for both self-CD1 and foreign lipid Ags. The dual reactivity of these CD1-restricted cells suggests that the capacity for rapid responses to inflammatory stimuli without memory coexists with the capacity for strong Ag-specific responses and the generation of memory in vivo. PMID- 16272287 TI - Interaction between human NK cells and bone marrow stromal cells induces NK cell triggering: role of NKp30 and NKG2D receptors. AB - In this study we have analyzed the interaction between in vitro cultured bone marrow stromal cells (BMSC) and NK cells. Ex vivo-isolated NK cells neoexpressed the activation Ag CD69 and released IFN-gamma and TNF-alpha upon binding with BMSC. Production of these proinflammatory cytokines was dependent on ligation of ICAM1 expressed on BMSC and its receptor LFA1 on NK cells. Furthermore, the NKp30, among natural cytotoxicity receptors, appeared to be primarily involved in triggering NK cells upon interaction with BMSC. Unexpectedly, autologous IL-2 activated NK cells killed BMSC. Again, LFA1/ICAM1 interaction plays a key role in NK/BMSC interaction; this interaction is followed by a strong intracellular calcium increase in NK cells. More importantly, NKG2D/MHC-I-related stress inducible molecule A and/or NKG2D/UL-16 binding protein 3 engagement is responsible for the delivery of a lethal hit. It appears that HLA-I molecules do not protect BMSC from NK cell-mediated injury. Thus, NK cells, activated upon binding with BMSC, may regulate BMSC survival. PMID- 16272289 TI - Evaluation of OX40 ligand as a costimulator of human antiviral memory CD8 T cell responses: comparison with B7.1 and 4-1BBL. AB - CTL are important effectors of antiviral immunity. Designing adjuvants that can induce strong cytotoxic T cell responses in humans would greatly improve the effectiveness of an antiviral vaccination or therapeutic strategy. Recent evidence suggests that, in addition to its well-established role in costimulation of CD4 T cell responses, OX40L (CD134) can directly costimulate mouse CD8 T cells. In this study, we evaluated the role of OX40L in costimulation of human antiviral CD8 T cell responses and compared it with two other important costimulators, B7.1 (CD80) and 4-1BBL (CD137L). Delivery of OX40L to human monocytes using a recombinant replication-defective adenovirus led to greater expansion, up-regulation of perforin, enhanced cytolytic activity, and increased numbers of IFN-gamma- and TNF-alpha-producing antiviral memory CD8 T cells in cultures of total T cells. Synergistic or additive effects were observed when OX40L costimulation was combined with 4-1BBL (CD137L) or B7.1 (CD80) costimulation. In total T cell cultures, at low Ag dose, 4-1BBL provided the most potent costimulus for influenza-specific CD8 T cell expansion, followed by B7.1 (CD80) and then OX40L. For isolated CD8 T cells, 4-1BBL was also the most consistent costimulator, followed by B7.1. In contrast, OX40L showed efficacy in direct activation of memory CD8 T cells in only one of seven donors. Thus, OX40L costimulates human antiviral memory CD8 T cell responses largely through indirect effects and can enhance anti-influenza, anti-EBV, and anti-HIV responses, particularly in combination with 4-1BBL or B7. PMID- 16272288 TI - Morphine induces CD4+ T cell IL-4 expression through an adenylyl cyclase mechanism independent of the protein kinase A pathway. AB - Impaired host defense mechanisms after major operative procedures and trauma are recognized as important factors in the development of infectious complication. Trauma is associated with impaired cellular immunity and CD4+ T cell Th2 differentiation. We have previously implicated morphine treatment as a possible mechanism for Th2 differentiation after injury. In this investigation we first establish that morphine treatment in vivo results in Th2 differentiation and that this effect is mediated through a naltrexone-sensitive opioid receptor. We investigated the intracellular mechanism by which morphine controls CD4+ T cell differentiation and demonstrate that morphine treatment in vitro 1) increases anti CD3/CD28 Ab-induced CD4+ T cell IL-4 protein synthesis, IL-4 mRNA, and GATA 3 mRNA accumulation through a pertussis toxin-sensitive receptor; 2) results in a dose-dependent increase in anti-CD3/CD28 Ab-induced CD4+ T cell cytoplasmic cAMP concentration; and 3) increases the forskolin-stimulated cytoplasmic cAMP level through a pertussis toxin-sensitive receptor. We also demonstrate that chronic morphine treatment increases anti-CD3/CD28 Ab-induced IL-4 promoter activity and IL-4 immunoprotein expression through a p38 MAPK-dependent, but protein kinase A- and Erk1/Erk2-independent, mechanism. PMID- 16272290 TI - Enhancement of HIV-specific CD8 T cell responses by dual costimulation with CD80 and CD137L. AB - HIV-specific CD8 T cell responses are defective in chronic HIV infection. In this study, we report that costimulation with either CD137L (4-1BBL) or CD80 (B7.1) enhanced the Ag-specific expansion and acquisition of effector function by HIV specific memory CD8 T cells. Ag-specific T cells from recently infected donors showed maximal expansion with single costimulatory molecules. Dual costimulation of T cells from recently infected donors or from healthy donors responding to influenza epitopes led to enhanced responses when the accumulation of cytokines was measured. However, accumulation of regulatory cytokines, particularly IFN gamma, led to inhibition of further Ag-specific CD8 T cell expansion in the cultures. This inhibition was relieved by neutralization of IFN-gamma or of IFN gamma, TNF, and IL-10. Thus, strong costimulation of T cells in vitro can lead to induction of regulatory cytokines at levels that limit further T cell expansion. In marked contrast, T cells from long-term (>4 years) infected HIV+ donors exhibited reduced Ag-specific CD8 T cell expansion, reduced CD4 T cell responses, and minimal cytokine accumulation. Dual costimulation with both 4-1BBL and B7.1 enhanced responses of T cells from long-term infected subjects to a level similar to that obtained with T cells from early in HIV infection. Experiments with purified CD8 T cells showed that B7.1 and 4-1BBL could act directly and synergistically on CD8 T cells. Taken together, these data suggest that 4-1BBL and B7.1 have additive or synergistic effects on HIV-specific CD8 T cell responses and represent a promising combination for therapeutic vaccination for HIV. PMID- 16272291 TI - TLR-dependent IL-4 production by invariant Valpha14+Jalpha18+ NKT cells to initiate contact sensitivity in vivo. AB - LPS stimulated B-1 cell polyclonal in vivo IgM responses depend on IL-4 release by invariant Valpha14+Jalpha18+ NKT (iNKT) cells. The IgM Abs can recruit effector T cells to mediate contact sensitivity. LPS activates the B-1 cell response just 1 day later, and depends on CD1d, iNKT cells, IL-4, TLR4, and MyD88. LPS in vivo and in vitro stimulates rapid preferential production of IL-4 in hepatic iNKT cells within 2 h. TLR4 were demonstrated in iNKT cells by flow cytometry and functional studies. Thus, innate microbial stimulation via TLR can activate iNKT cell and B-1 cell collaboration. The result is polyclonal IgM Ab responses capable of recruiting Ag-specific T cells into tissues. This may be involved in the promotion of autoimmunity by infectious agents. PMID- 16272293 TI - Control of genotypic allelic inclusion through TCR surface expression. AB - To gain insight into the molecular causes and functional consequences of allelic inclusion of TCR alpha-chains, we develop a computational model for thymocyte selection in which the signal that determines cell fate depends on surface expression. Analysis of receptor pairs on selected dual TCR cells reveals that allelic inclusion permits both autoreactive TCR and receptors not in the single TCR cell repertoire to be selected. However, in comparison with earlier theoretical studies, relatively few dual TCR cells display receptors with high avidity for thymic ligands because their alpha-chains compete aggressively for the beta-chain, which hinders rescue from clonal deletion. This feature of the model makes clear that allelic inclusion does not in itself compromise central tolerance. A specific experiment based on modulation of TCR surface expression levels is proposed to test the model. PMID- 16272292 TI - Cyclin T1 expression is regulated by multiple signaling pathways and mechanisms during activation of human peripheral blood lymphocytes. AB - Stimulation of primary human T lymphocytes results in up-regulation of cyclin T1 expression, which correlates with phosphorylation of the C-terminal domain of RNA polymerase II (RNAP II). Up-regulation of cyclin T1 and concomitant stabilization of cyclin-dependent kinase 9 (CDK9) may facilitate productive replication of HIV in activated T cells. We report that treatment of PBLs with two mitogens, PHA and PMA, results in accumulation of cyclin T1 via distinct mechanisms. PHA induces accumulation of cyclin T1 mRNA and protein, which results from cyclin T1 mRNA stabilization, without significant change in cyclin T1 promoter activity. Cyclin T1 mRNA stabilization requires the activation of both calcineurin and JNK because inhibition of either precludes cyclin T1 accumulation. In contrast, PMA induces cyclin T1 protein up-regulation by stabilizing cyclin T1 protein, apparently independently of the proteasome and without accumulation of cyclin T1 mRNA. This process is dependent on Ca2+-independent protein kinase C activity but does not require ERK1/2 activation. We also found that PHA and anti-CD3 Abs induce the expression of both the cyclin/CDK complexes involved in RNAP II C-terminal domain phosphorylation and the G1-S cyclins controlling cell cycle progression. In contrast, PMA alone is a poor inducer of the expression of G1-S cyclins but often as potent as PHA in inducing RNAP II cyclin/CDK complexes. These findings suggest coordination in the expression and activation of RNAP II kinases by pathways that independently stimulate gene expression but are insufficient to induce S phase entry in primary T cells. PMID- 16272294 TI - Functional analysis of B and T lymphocyte attenuator engagement on CD4+ and CD8+ T cells. AB - T cell activation can be profoundly altered by coinhibitory and costimulatory molecules. B and T lymphocyte attenuator (BTLA) is a recently identified inhibitory Ig superfamily cell surface protein found on lymphocytes and APC. In this study we analyze the effects of an agonistic anti-BTLA mAb, PK18, on TCR mediated T cell activation. Unlike many other allele-specific anti-BTLA mAb we have generated, PK18 inhibits anti-CD3-mediated CD4+ T cell proliferation. This inhibition is not dependent on regulatory T cells, nor does the Ab induce apoptosis. Inhibition of T cell proliferation correlates with a profound reduction in IL-2 secretion, although this is not the sole cause of the block of cell proliferation. In contrast, PK18 has no effect on induction of the early activation marker CD69. PK18 also significantly inhibits, but does not ablate, IL 2 secretion in the presence of costimulation as well as reduces T cell proliferation under limiting conditions of activation in the presence of costimulation. Similarly, PK18 inhibits Ag-specific T cell responses in culture. Interestingly, PK18 is capable of delivering an inhibitory signal as late as 16 h after the initiation of T cell activation. CD8+ T cells are significantly less sensitive to the inhibitory effects of PK18. Overall, BTLA adds to the growing list of cell surface proteins that are potential targets to down-modulate T cell function. PMID- 16272295 TI - CD11chigh dendritic cell ablation impairs lymphopenia-driven proliferation of naive and memory CD8+ T cells. AB - The peripheral lymphocyte pool size is governed by homeostatic mechanisms. Thus, grafted T cells expand and replenish T cell compartments in lymphopenic hosts. Lymphopenia-driven proliferation of naive CD8+ T cells depends on self peptide/MHC class I complexes and the cytokine IL-7. Lymphopenia-driven proliferation and maintenance of memory CD8+ T cells are MHC independent, but are believed to require IL-7 and contact with a bone marrow-derived cell that presents the cytokine IL-15 by virtue of its high affinity receptor (IL 15Ralpha). In this study we show that optimal spontaneous proliferation of grafted naive and memory CD8+ T cells in mice rendered lymphopenic through gene ablation or irradiation requires the presence of CD11chigh dendritic cells. Our results suggest a dual role of CD11chigh dendritic cells as unique APC and cytokine-presenting cells. PMID- 16272296 TI - Phagocytosis of hemozoin enhances matrix metalloproteinase-9 activity and TNF alpha production in human monocytes: role of matrix metalloproteinases in the pathogenesis of falciparum malaria. AB - Matrix metalloproteinase-9 (MMP-9), secreted by activated monocytes, degrades matrix proteins, disrupts basal lamina, and activates TNF-alpha from its precursors. In turn, TNF-alpha enhances synthesis of MMP-9 in monocytes. We show here that trophozoite-parasitized RBCs/hemozoin-fed adherent human monocytes displayed increased MMP-9 activity and protein/mRNA expression, produced TNF alpha time-dependently, and showed higher matrix invasion ability. MMP-9 activation was specific for trophozoite/hemozoin-fed monocytes, was dependent on TNF-alpha production, and abrogated by anti-TNF-alpha Ab and by a specific inhibitor of MMP-9/MMP-13 activity. Hemozoin-induced enhancement of MMP-9 and TNF alpha production would have a 2-fold effect: to start and feed a cyclic reinforcement loop in which hemozoin enhances production of TNF-alpha, which in turn induces both activation of MMP-9 and shedding of TNF-alpha into the extracellular compartment; and, second, to disrupt the basal lamina of endothelia. Excess production of TNF-alpha and disruption of the basal lamina with extravasation of blood cells into perivascular tissues are hallmarks of severe malaria. Pharmacological inhibition of MMP-9 may offer a new chance to control pathogenic mechanisms in malaria. PMID- 16272297 TI - The murine B cell repertoire is severely selected against endogenous cellular prion protein. AB - Abs to the prion protein (PrP) can protect against experimental prion infections, but efficient Ab responses are difficult to generate because PrP is expressed on many tissues and induces a strong tolerance. We previously showed that immunization of wild-type mice with PrP peptides and CpG oligodeoxynucleic acid overcomes tolerance and induces cellular and humoral responses to PrP. In this study, we compared Ab and T cell repertoires directed to PrP in wild-type and PrP knockout (Prnp o/o) C57BL/6 mice. Animals were immunized with mouse PrP-plasmid DNA or with 30-mer overlapping peptides either emulsified in CFA or CpG/IFA. In Prnp o/o mice, Abs raised by PrP-plasmid DNA immunization recognized only N terminal PrP peptides; analyses of Ab responses after PrP peptide/CFA immunization allowed us to identify six distinct epitopes, five of which were also recognized by Abs raised by PrP peptides/CpG. By contrast, in wild-type mice, no Ab response was detected after PrP-plasmid DNA or peptide/CFA immunization. However, when using CpG, four C-terminal peptides induced Abs specific for distinct epitopes. Importantly, immune sera from Prnp o/o but not from wild-type mice bound cell surface PrP. Abs of IgG1 and IgG2b subclasses predominated in Prnp o/o mice while the strongest signals were for IgG2b in wild type mice. Most anti-PrP Th cells were directed to a single epitope in both Prnp o/o and wild-type mice. We conclude that endogenous PrPC expression profoundly affects the Ab repertoire as B cells reactive for epitopes exposed on native PrPC are strongly tolerized. Implications for immunotherapy against prion diseases are discussed. PMID- 16272298 TI - Thymus-dependent memory phenotype CD8 T cells in naive B6.H-2Kb-/-Db-/- animals mediate an antigen-specific response against Listeria monocytogenes. AB - B6.H-2Kb-/-Db-/- (DKO) mice have greatly reduced numbers of mature CD8alphabeta T cells in their periphery. However, these non-class Ia-selected CD8alphabeta T cells are able to mediate immune responses to a number of pathogens. Approximately 60% of the CD8alphabeta T cells in the spleen and peripheral lymph nodes of naive DKO mice display a memory (CD44high) phenotype. To investigate the origins of these non-class Ia-selected CD8alphabetaCD44high cells, we traced the phenotype of recent thymic emigrants and found that most were CD44low. We also determined whether their appearance was thymus dependent and found that only a small percentage of non-class Ia-selected CD8alphabetaCD44high cells develop in a thymus-independent pathway. Functionally, CD8alphabetaCD44high cells from DKO mice are able to secrete IFN-gamma in response to IL-12 and IL-18 in the absence of cognate Ag. When challenged with anti-CD3 in vivo, nearly half of these cells produce IFN-gamma within 3 h. When purified CD8alphabetaCD44high cells from Thy1.2.DKO mice were transferred into Thy1.1 DKO recipients and then challenged with Listeria monocytogenes, an Ag-specific anti-L. monocytogenes response was observed 6 days later. Our data suggest that non-class Ia-selected CD8alphabetaCD44high cells in naive animals can respond rapidly to Ag and play a role in the innate as well as the early phase of the acquired immune response. PMID- 16272299 TI - CD73 and Ly-6A/E distinguish in vivo primed but uncommitted mouse CD4 T cells from type 1 or type 2 effector cells. AB - Primed CD4 T cells may develop into effector T cells such as Th1 and Th2, or remain uncommitted as Th primed precursor (Thpp) cells that can subsequently differentiate into Th1 and Th2 cells. Although mouse Thpp-like cells have also been identified among spleen and particularly lymph node cells, further characterization of these cells has been difficult without a defining cell surface marker. Using Affymetrix GeneChips followed by FACS analysis, we found that in vitro-derived Thpp cells expressed CD73 but not Ly-6A/E, whereas Th1 and Th2 cells showed the reciprocal pattern. CD73+ Ly6A/E- memory CD4 T cells were identified in normal C57BL/6 mice, and the proportion of these cells was highest in lymph nodes, lower in spleens, and lowest in the lungs. These cells produced IL-2 and MIP-1alpha, but much less IL-4 and IFN-gamma than CD73- Ly6A/E+ cells. Similar results were obtained with additional Ly-6.2 mouse strains, but not Ly 6.1 strains. Restimulation of Thpp-like CD73+ Ly-6A/E- cells in Th1- or Th2 polarizing conditions induced differentiation into populations producing mainly IFN-gamma or mainly IL-4, respectively. In contrast, the effector-like CD73- Ly 6A/E+ population was more committed, and continued to produce both IL-4 and IFN gamma in both conditions. CD73 and Ly-6A/E expression therefore identify a population of Thpp-like cells in C57BL/6 mice and at least some other Ly-6.2 mice. PMID- 16272301 TI - DM peptide-editing function leads to immunodominance in CD4 T cell responses in vivo. AB - DM functions as a peptide editor for MHC class II-bound peptides. We examined the hypothesis that DM peptide editing plays a key role in focusing the in vivo CD4 T cell responses against complex pathogens and protein Ags to only one, or at most a few, immunodominant peptides. Most CD4 T cells elicited in the wild-type BALB/c (H-2d) mice infected with Leishmania major predominantly recognize a single epitope 158-173 within Leishmania homologue of activated receptor for c-kinase (LACK), as is the case when these mice are immunized with rLACK. Using DM deficient (DM-/-) H-2d mice, we now show that in the absence of DM, the in vivo CD4 T cell responses to rLACK are skewed away from the immunodominant epitopes and are diversified to include two novel epitopes (LACK 33-48 and 261-276). DM-/- B10.BR (H-2k) mice showed similar results. These results constitute the first demonstration of the role of DM peptide editing in sculpting the specificity and immunodominance in in vivo CD4 T cell responses. PMID- 16272300 TI - Pharmacological inhibition of endotoxin responses is achieved by targeting the TLR4 coreceptor, MD-2. AB - The detection of Gram-negative LPS depends upon the proper function of the TLR4 MD-2 receptor complex in immune cells. TLR4 is the signal transduction component of the LPS receptor, whereas MD-2 is the endotoxin-binding unit. MD-2 appears to activate TLR4 when bound to TLR4 and ligated by LPS. Only the monomeric form of MD-2 was found to bind LPS and only monomeric MD-2 interacts with TLR4. Monomeric MD-2 binds TLR4 with an apparent Kd of 12 nM; this binding avidity was unaltered in the presence of endotoxin. E5564, an LPS antagonist, appears to inhibit cellular activation by competitively preventing the binding of LPS to MD-2. Depletion of endogenous soluble MD-2 from human serum, with an immobilized TLR4 fusion protein, abrogated TLR4-mediated LPS responses. By determining the concentration of added-back MD-2 that restored normal LPS responsiveness, the concentration of MD-2 was estimated to be approximately 50 nM. Similarly, purified TLR4-Fc fusion protein, when added to the supernatants of TLR4 expressing cells in culture, inhibited the interaction of MD-2 with TLR4, thus preventing LPS stimulation. The ability to inhibit the effects of LPS as a result of the binding of TLR4-Fc or E5564 to MD-2 highlights MD-2 as the logical target for drug therapies designed to pharmacologically intervene against endotoxin induced disease. PMID- 16272302 TI - Disruption of MHC class II-restricted antigen presentation by vaccinia virus. AB - Vaccinia virus (VV), currently used in humans as a live vaccine for smallpox, can interfere with host immunity via several discrete mechanisms. In this study, the effect of VV on MHC class II-mediated Ag presentation was investigated. Following VV infection, the ability of professional and nonprofessional APC to present Ag and peptides to CD4+ T cells was impaired. Viral inhibition of class II Ag presentation could be detected within 1 h, with diminished T cell responses dependent upon the duration of APC infection and virus titer. Exposure of APC to replication-deficient virus also diminished class II Ag presentation. Virus infection of APC perturbed Ag presentation by newly synthesized and recycling class II molecules, with disruptions in both exogenous and cytoplasmic Ag presentation. Virus-driven expression of an endogenous Ag, failed to restore T cell responsiveness specific for this Ag in the context of MHC class II molecules. Yet, both class II protein steady-state and cell surface expression were not altered by VV. Biochemical and functional analysis revealed that VV infection directly interfered with ligand binding to class II molecules. Together, these observations suggest that disruption of MHC class II-mediated Ag presentation may be one of multiple strategies VV has evolved to escape host immune surveillance. PMID- 16272303 TI - Stepwise differentiation of CD4 memory T cells defined by expression of CCR7 and CD27. AB - To study the steps in the differentiation of human memory CD4 T cells, we characterized the functional and lineage relationships of three distinct memory CD4 subpopulations distinguished by their expression of the cysteine chemokine receptor CCR7 and the TNFR family member CD27. Using the combination of these phenotypic markers, three populations were defined: the CCR7+CD27+, the CCR7 CD27+, and the CCR7-CD27- population. In vitro stimulation led to a stepwise differentiation from naive to CCR7+CD27+ to CCR7-CD27+ to CCR7-CD27-. Telomere length in these subsets differed significantly (CCR7+CD27+ > CCR7-CD27+ > CCR7 CD27-), suggesting that these subsets constituted a differentiative pathway with progressive telomere shortening reflecting antecedent in vivo proliferation. The in vitro proliferative response of these populations declined, and their susceptibility to apoptosis increased progressively along this differentiation pathway. Cytokine secretion showed a differential functional capacity of these subsets. High production of IL-10 was only observed in CCR7+CD27+, whereas IFN gamma was produced by CCR7-CD27+ and to a slightly lesser extent by CCR7-CD27- T cells. IL-4 secretion was predominantly conducted by CCR7-CD27- memory CD4 T cells. Thus, by using both CCR7 and CD27, distinct maturational stages of CD4 memory T cells with different functional activities were defined. PMID- 16272304 TI - The src homology 2 domain-containing tyrosine phosphatase 2 regulates primary T dependent immune responses and Th cell differentiation. AB - The src homology 2 domain-containing tyrosine phosphatase 2 (SHP2) plays an important role in development and in growth factor receptor signaling pathways, yet little is known of its role in the immune system. We generated mice expressing a dominant-negative version of the protein, SHP2(CS), specifically in T cells. In SHP2(CS) mice, T cell development appears normal with regard to both negative and positive selection. However, SHP2(CS) T cells express higher levels of activation markers, and aged mice have elevated serum Abs. This is associated with a marked increase in IL-4, IL-5, and IL-10 secretion by SHP2(CS) T cells in vitro. In addition, primary thymus-dependent B cell responses are deficient in SHP2(CS) mice. We show that whereas TCR-induced linker for activation of T cells phosphorylation is defective, CTLA-4 and programmed death-1 signaling are not affected by SHP2(CS) expression. Our results suggest that a key action of wild type SHP2 is to suppress differentiation of T cells to the Th2 phenotype. PMID- 16272305 TI - IL-1beta-mediated proinflammatory responses are inhibited by estradiol via down regulation of IL-1 receptor type I in uterine epithelial cells. AB - The objective of this study was to examine the effects of sex hormones on IL 1beta-mediated responses by uterine epithelial cells. The mRNA expression and secretion of human beta-defensin-2 and CXCL8 by uterine epithelial cells was examined following stimulation with IL-1beta in the presence of estradiol or progesterone. Estradiol inhibited the IL-1beta-mediated mRNA expression and secretion of human beta-defensin-2 and CXCL8 by uterine epithelial cells while progesterone had no effect. Inhibition of the IL-1beta-mediated response by estradiol was dose dependent, with maximal inhibition observed using 10(-7) to 10(-10) M, and was shown to be mediated through the estrogen receptor because addition of a pure estrogen receptor antagonist abrogated this effect. The mechanism by which estradiol inhibits IL-1beta-mediated responses by uterine epithelial cells appears to be the down-modulation of the IL-1R type I, thereby reducing the uterine epithelial cell's ability to respond to IL-1beta. These results suggest that the inhibitory effect of estradiol on IL-1beta-mediated inflammatory responses by uterine epithelial cells indicates a link between the endocrine and immune systems and may be crucial for dampening proinflammatory responses during the time of ovulation or pregnancy. PMID- 16272306 TI - Rab5 and Rab7, but not ARF6, govern the early events of HIV-1 infection in polarized human placental cells. AB - Trophoblasts, the structural cells of the placenta, are thought to play a determinant role in in utero HIV type 1 (HIV-1) transmission. We have accumulated evidence suggesting that HIV-1 infection of these cells is associated with uptake by an unusual clathrin/caveolae-independent endocytic pathway and that endocytosis is followed by trafficking through multiple organelles. Furthermore, part of this trafficking involves the transit of HIV-1 from transferrin-negative to EEA1 and transferrin-positive endosomes, suggesting a merger from nonclassical to classical endocytic pathways in these cells. In the present article, the relationship between the presence of HIV-1 within specific endosomes and infection was studied. We demonstrate that viral infection is virtually lost when endosome inhibitors are added shortly after exposure to HIV-1. Thus, contrary to what is seen in CD4+ T lymphocytes, the initial presence of HIV-1 within the endosomes is mandatory for infection to take place. Importantly, this process is independent of the viral envelope proteins gp120 and gp41. The Rab family of small GTPases coordinates the vesicular transport between the different endocytic organelles. Experiments performed with various expression vectors indicated that HIV-1 infection in polarized trophoblasts relies on Rab5 and Rab7 without the contribution of Arf6 or Rab11. Furthermore, we conclude that Rab5 drives movements from raft-rich region to early endosomes, and this transit is required for subsequently reaching late endosomes via Rab7. This complex trafficking is mandatory for HIV-1 infection to proceed in human polarized trophoblasts. PMID- 16272307 TI - Prostaglandin D2 causes preferential induction of proinflammatory Th2 cytokine production through an action on chemoattractant receptor-like molecule expressed on Th2 cells. AB - PGD2, produced by mast cells, has been detected in high concentrations at sites of allergic inflammation. It can stimulate vascular and other inflammatory responses by interaction with D prostanoid receptor (DP) and chemoattractant receptor-like molecule expressed on Th2 cells (CRTH2) receptors. A significant role for PGD2 in mediating allergic responses has been suggested based on the observation that enhanced eosinophilic lung inflammation and cytokine production is apparent in the allergen-challenged airways of transgenic mice overexpressing human PGD2 synthase, and PGD2 can enhance Th2 cytokine production in vitro from CD3/CD28-costimulated Th2 cells. In the present study, we investigated whether PGD2 has the ability to stimulate Th2 cytokine production in the absence of costimulation. At concentrations found at sites of allergic inflammation, PGD2 preferentially elicited the production of IL-4, IL-5, and IL-13 by human Th2 cells in a dose-dependent manner without affecting the level of the anti inflammatory cytokine IL-10. Gene transcription peaked within 2 h, and protein release peaked approximately 8 h after stimulation. The effect of PGD2 was mimicked by the selective CRTH2 agonist 13,14-dihydro-15-keto-PGD2 but not by the selective DP agonist BW245C, suggesting that the stimulation is mediated by CRTH2 and not DP. Ramatroban, a dual CRTH2/thromboxane-like prostanoid receptor antagonist, markedly inhibited Th2 cytokine production induced by PGD2, while the selective thromboxane-like prostanoid receptor antagonist SQ29548 was without effect. These data suggest that PGD2 preferentially up-regulates proinflammatory cytokine production in human Th2 cells through a CRTH2-dependent mechanism in the absence of any other costimulation and highlight the potential utility of CRTH2 antagonists in the treatment of allergic diseases. PMID- 16272308 TI - CD1d-independent developmental acquisition of prompt IL-4 gene inducibility in thymus CD161(NK1)-CD44lowCD4+CD8- T cells is associated with complementarity determining region 3-diverse and biased Vbeta2/Vbeta7/Vbeta8/Valpha3.2 T cell receptor usage. AB - Among Ag-inexperienced naive T cells, the CD1d-restricted NKT cell that uses invariant TCR-alpha-chain is the most widely studied cell capable of prompt IL-4 inducibility. We show in this study that thymus CD161-CD44lowCD4+CD8- T cells promptly produce IL-4 upon TCR stimulation, a response that displays biased Vbeta(2/7/8) and Valpha3.2 TCR usage. The association of Vbeta family bias and IL 4 inducibility in thymus CD161-CD44lowCD4+CD8- T cells is found for B6, B10, BALB/c, CBA, B10.A(4R), and ICR mouse strains. Despite reduced IL-4 inducibility, there is a similarly biased Vbeta(2/7/8) TCR usage by IL-4 inducibility+ spleen CD161-CD44lowCD4+CD8- T cells. Removal of alpha-galacotosylceramide/CD1d-binding cells from CD161-CD44lowCD4+CD8- thymocytes does not significantly affect their IL-4 inducibility. The development of thymus CD161-CD44lowCD4+CD8- T cells endowed with IL-4 inducibility and their associated use of Vbeta(2/7/8) are beta2 microglobulin-, CD1d-, and p59fyn-independent. Thymus CD161-CD44lowCD4+CD8- T cells produce low and no IFN-gamma inducibility in response to TCR stimulation and to IL-12 + IL-18, respectively, and they express diverse complementarity determining region 3 sequences for both TCR-alpha- and -beta-chains. Taken together, these results demonstrate the existence of a NKT cell distinct, TCR repertoire diverse naive CD4+ T cell subset capable of prompt IL-4 inducibility. This subset has the potential to participate in immune response to a relatively large number of Ags. The more prevalent nature of this unique T cell subset in the thymus than the periphery implies roles it might play in intrathymic T cell development and may provide a framework upon which mechanisms of developmentally regulated IL-4 gene inducibility can be studied. PMID- 16272309 TI - The cooperation between two CD4 T cells induces tumor protective immunity in MUC.1 transgenic mice. AB - Immunity and tumor protection in mice transgenic for human MUC.1, a glycoprotein expressed in the majority of cancers of epithelial origin in humans, were induced by vaccination with B lymphocytes genetically programmed to activate MUC.1 specific CD4 T cells. Their activation required a functional cooperation between two Th cells, one specific for a self (MUC.1) and the other for a nonself T cell determinant. The immunological switch provided by Th-Th cooperation was sufficient to induce MUC.1-specific CD4 and CD8 T cell responses in MUC.1 transgenic mice, and protect them permanently from tumor growth. CD4 T cells specific for MUC.1 lacked cytolytic function, but produced IFN-gamma upon restimulation with Ag. We conclude that immunity against tumor self-Ags and tumor protection can be regulated exploiting an inherent property of the immune system. PMID- 16272310 TI - Functional analysis of HIV type 1 Nef reveals a role for PAK2 as a regulator of cell phenotype and function in the murine dendritic cell line, DC2.4. AB - The HIV-1 Nef protein plays a critical role in viral pathogenesis. Nef has been shown to modulate dendritic cell (DC) function, in particular perturbing their ability to present Ag. To further characterize the effects of Nef on DCs, we established a panel of transfectants of the murine DC line, DC2.4, stably expressing differing levels of either wild-type Nef, or a number of Nef mutants lacking key functional motifs. Transfectants expressing increasing levels of wild type Nef demonstrated a dose-dependent shrinkage and loss of dendrites. Nef expression levels also correlated with increased proliferative ability but did not confer resistance to proapoptotic stimuli. Importantly, Nef expression resulted in an impairment of Ag presentation to T cells correlating with a reduction in the cell surface expression of molecules involved in Ag presentation such as MHC class I, CD80/86, and ICAM-1. Nef expression also rendered DC2.4 cells resistant to the maturation stimulus provided by an anti-CD40 Ab. Mutations in either the myristoylation site or Src homology 3-domain binding polyproline motif of Nef abolished these effects. Previous studies had shown that these mutations also abolished the ability of Nef to activate the p21-activated kinase, PAK2. Consistent with this, stable expression of constitutively active PAK2 in DC2.4 mimicked the effects of Nef. We conclude that Nef, acting via activation of PAK2, inhibits both DC maturation and Ag presentation. These data have clear implications for the role of Nef in early stages of HIV-1 infection and validate Nef as a valid target for development of antiviral chemotherapeutics. PMID- 16272311 TI - Differential expression of IFN regulatory factor 4 gene in human monocyte-derived dendritic cells and macrophages. AB - In vitro human monocyte differentiation to macrophages or dendritic cells (DCs) is driven by GM-CSF or GM-CSF and IL-4, respectively. IFN regulatory factors (IRFs), especially IRF1 and IRF8, are known to play essential roles in the development and functions of macrophages and DCs. In the present study, we performed cDNA microarray and Northern blot analyses to characterize changes in gene expression of selected genes during cytokine-stimulated differentiation of human monocytes to macrophages or DCs. The results show that the expression of IRF4 mRNA, but not of other IRFs, was specifically up-regulated during DC differentiation. No differences in IRF4 promoter histone acetylation could be found between macrophages and DCs, suggesting that the gene locus was accessible for transcription in both cell types. Computer analysis of the human IRF4 promoter revealed several putative STAT and NF-kappaB binding sites, as well as an IRF/Ets binding site. These sites were found to be functional in transcription factor-binding and chromatin immunoprecipitation experiments. Interestingly, Stat4 and NF-kappaB p50 and p65 mRNAs were expressed at higher levels in DCs as compared with macrophages, and enhanced binding of these factors to their respective IRF4 promoter elements was found in DCs. IRF4, together with PU.1, was also found to bind to the IRF/Ets response element in the IRF4 promoter, suggesting that IRF4 protein provides a positive feedback signal for its own gene expression in DCs. Our results suggest that IRF4 is likely to play an important role in myeloid DC differentiation and gene regulatory functions. PMID- 16272312 TI - Sphingosine kinase 1 is a negative regulator of CD4+ Th1 cells. AB - CD4+ Th1 cells produce IFN-gamma, TNF-alpha, and IL-2. These Th1 cytokines play critical roles in both protective immunity and inflammatory responses. In this study we report that sphingosine kinase 1 (SPHK1), but not SPHK2, is highly expressed in DO11.10 Th1 cells. The expression of SPHK1 in Th1 cells requires TCR signaling and new protein synthesis. SPHK1 phosphorylates sphingosine to form sphingosine-1-phosphate. Sphingosine-1-phosphate plays important roles in inhibition of apoptosis, promotion of cell proliferation, cell migration, calcium mobilization, and activation of ERK1/2. When SPHK1 expression was knocked down by SPHK1 short interfering RNA, the production of IL-2, TNF-alpha, and IFN-gamma by Th1 cells in response to TCR stimulation was enhanced. Consistently, overexpression of dominant-negative SPHK1 increased the production of IL-2, TNF alpha, and IFN-gamma in Th1 cells. Furthermore, overexpression of SPHK1 in Th1 and Th0 cells decreased the expression of IL-2, TNF-alpha, and IFN-gamma. Several chemokines, including Th2 chemokines CCL17 and CCL22, were up-regulated by SPHK1 short interfering RNA and down-regulated by overexpression of SPHK1. We also showed that Th2 cells themselves express CCL17 and CCL22. Finally, we conclude that SPHK1 negatively regulates the inflammatory responses of Th1 cells by inhibiting the production of proinflammatory cytokines and chemokines. PMID- 16272313 TI - A high-affinity natural autoantibody from human cord blood defines a physiologically relevant epitope on the FcepsilonRIalpha. AB - Natural Abs represent the indigenous immune repertoire and are thus present at birth and persist throughout life. Previously, human autoantibodies to the alpha domain of the high-affinity IgE receptor (FcepsilonRIalpha) have been isolated from Ab libraries derived from normal donors and patients with chronic urticaria. To investigate whether these anti-FcepsilonRIalpha Abs are present in the germline repertoire, we constructed a phage Fab display library from human cord blood, which represents the naive immune repertoire before exposure to exogenous Ags. All isolated clones specific to the FcepsilonRIalpha had the same sequence. This single IgM Ab, named CBMalpha8, was strictly in germline configuration and had high affinity and functional in vitro anaphylactogenic activity. Inhibition experiments indicated an overlapping epitope on the FcepsilonRIalpha recognized by both CBMalpha8 and the previously isolated anti-FcepsilonRIalpha Abs from autoimmune and healthy donors. This common epitope on FcepsilonRIalpha coincides with the binding site for IgE. Affinity measurements demonstrated the presence of Abs showing CBMalpha8-like specificity, but with a significantly lower affinity in i.v. Ig, a therapeutic multidonor IgG preparation. We propose a hypothesis of escape mutants, whereby the resulting lower affinity IgG anti-FcepsilonRIalpha Abs are rendered less likely to compete with IgE for binding to FcepsilonRIalpha. PMID- 16272314 TI - Activation and inactivation of antiviral CD8 T cell responses during murine pneumovirus infection. AB - Pneumonia virus of mice (PVM) is a natural pathogen of mice and has been proposed as a tractable model for the replication of a pneumovirus in its natural host, which mimics human infection with human respiratory syncytial virus (RSV). PVM infection in mice is highly productive in terms of virus production compared with the situation seen with RSV in mice. Because RSV suppresses CD8 T cell effector function in the lungs of infected mice, we have investigated the nature of PVM induced CD8 T cell responses to study pneumovirus-induced T cell responses in a natural virus-host setting. PVM infection was associated with a massive influx of activated CD8 T cells into the lungs. After identification of three PVM-specific CD8 T cell epitopes, pulmonary CD8 T cell responses were enumerated. The combined frequency of cytokine-secreting CD8 T cells specific for the three epitopes was much smaller than the total number of activated CD8 T cells. Furthermore, quantitation of the CD8 T cell response against one of these epitopes (residues 261-270 from the phosphoprotein) by MHC class I pentamer staining and by in vitro stimulation followed by intracellular IFN-gamma and TNF-alpha staining indicated that the majority of pulmonary CD8 specific for the P261 epitope were deficient in cytokine production. This deficient phenotype was retained up to 96 days postinfection, similar to the situation in the lungs of human RSV-infected mice. The data suggest that PVM suppresses T cell effector functions in the lungs. PMID- 16272315 TI - Leucine aminopeptidase is not essential for trimming peptides in the cytosol or generating epitopes for MHC class I antigen presentation. AB - To detect viral infections and tumors, CD8+ T lymphocytes monitor cells for the presence of antigenic peptides bound to MHC class I molecules. The majority of MHC class I-presented peptides are generated from the cleavage of cellular and viral proteins by the ubiquitin-proteasome pathway. Many of the oligopeptides produced by this process are too long to stably bind to MHC class I molecules and require further trimming for presentation. Leucine aminopeptidase (LAP) is an IFN inducible cytosolic aminopeptidase that can trim precursor peptides to mature epitopes and has been thought to play an important role in Ag presentation. To examine the role of LAP in generating MHC class I peptides in vivo, we generated LAP-deficient mice and LAP-deficient cell lines. These mutant mice and cells are viable and grow normally. The trimming of peptides in LAP-deficient cells is not reduced under basal conditions or after stimulation with IFN. Similarly, there is no reduction in presentation of peptides from precursor or full-length Ag constructs or in the overall supply of peptides from cellular proteins to MHC class I molecules even after stimulation with IFN. After viral infection, LAP deficient mice generate normal CTL responses to seven epitopes from three different viruses. These data demonstrate that LAP is not an essential enzyme for generating most MHC class I-presented peptides and reveal redundancy in the function of cellular aminopeptidases. PMID- 16272316 TI - Tissue-resident memory CD8+ T cells can be deleted by soluble, but not cross presented antigen. AB - Under noninflammatory conditions, both naive and central memory CD8 T cells can be eliminated in the periphery with either soluble peptide or cross-presented Ag. Here, we assess the tolerance susceptibility of tissue-resident memory CD8 T cells in mice to these two forms of tolerogen. Soluble peptide specifically eliminated the majority of memory CD8 cells present in both lymphoid and extralymphoid tissues including lung and liver, but was unable to reduce the number present in the CNS. In contrast, systemic cross-presentation of Ag by dendritic cells resulted in successful elimination of memory cells only from the spleen, with no significant reduction in the numbers of tissue-resident memory cells in the lung. The fact that tissue-resident memory cells were unable to access cross-presented Ag suggests that either the memory cells in the lung do not freely circulate out of the tissue, or that they circulate through a region in the spleen devoid of cross-presented Ag. Thus, although tissue-resident memory cells are highly susceptible to tolerance induction, both the form of tolerogen and location of the T cells can determine their accessibility to tolerogen and the degree to which they are successfully deleted from specific tissues. PMID- 16272317 TI - A single DH gene segment creates its own unique CDR-H3 repertoire and is sufficient for B cell development and immune function. AB - To test the contribution of individual D gene segments to B cell development and function, we used gene targeting to create mice that contain only DFL16.1 in the DH locus. We term this D-limited IgH allele DeltaD-DFL. Although the absolute number of IgM+IgD- B cells in the bone marrow was decreased, homozygous DeltaD DFL BALB/c mice contained normal numbers of IgM+IgD+ B cells in bone marrow and spleen and normal numbers of B1a, B1b, and B2 cells in the peritoneal cavity. Bone marrow IgM+IgD+ B cells express a CDR-H3 repertoire similar in length and amino acid composition to the DFL16.1 subset of the wild-type BALB/c repertoire but divergent from sequences that do not contain DFL16.1. This similarity in content is the product of both germline bias and somatic selection, especially in the transition to the mature IgM+IgD+ stage of development. Serum Ig concentrations and the humoral immune response to a T-dependent Ag ([4-hydroxy-3 nitrophenyl]acetyl hapten) were nearly identical to wild-type littermate controls. A greater variance in the immune response to the T-independent Ag (alpha(1-->3)-dextran) was observed in DeltaD-DFL homozygotes, with half of the mice exhibiting levels below the range exhibited by controls. Although limited to a repertoire specific to DFL16.1, the presence of a single DH gene segment of normal sequence was sufficient for development of normal numbers of mature B cells and for robust humoral immune function. PMID- 16272318 TI - RNA stability of the E2A-encoded transcription factor E47 is lower in splenic activated B cells from aged mice. AB - We have demonstrated previously that DNA binding and protein expression of the E2A-encoded transcription factor E47 are lower in nuclear extracts of activated splenic B cells from old mice. In the present study, we address how E47 protein expression is regulated in aging. Results herein show that E2A mRNA levels were decreased in stimulated splenic B cells from old as compared with young mice. RNA stability assays showed that the rate of E2A mRNA decay was accelerated in stimulated splenic B cells from old mice, but E47 protein degradation rates were comparable in young vs aged B cells, indicating that the regulation of E47 expression in activated splenic B cells occurs primarily by mRNA stability. The rates of decay of other mRNAs showed that the increased mRNA degradation in aged splenic activated B cells is not a general phenomenon but restricted to a subset of mRNAs. We next investigated the signal transduction pathways controlling E2A mRNA expression and stability and found that p38 MAPK regulates E2A mRNA expression through increased mRNA stability and is down-regulated in aged activated B cells. Results show that inhibition of p38 MAPK significantly reduces E2A mRNA stability in both young and old B cells, further stressing the role of p38 MAPK in E2A RNA stabilization. These studies demonstrate that the transcription factor E2A, critical for many aspects of B cell function, is regulated by a novel mechanism in aging. PMID- 16272319 TI - Dimerization of the major birch pollen allergen Bet v 1 is important for its in vivo IgE-cross-linking potential in mice. AB - In type I allergy, the cross-linking of membrane IgE on B lymphocytes and of cytophilic IgE on effector cells by their respective allergens are key events. For cross-linking two IgE molecules, allergens need at least two epitopes. On large molecules, these could be different epitopes in a multivalent, or identical epitopes in a symmetrical, fashion. However, the availability of epitopes may be limited on small allergens such as Bet v 1, the major birch pollen allergen. The present work analyzes whether dimerization is required for the cross-linking capacity of this allergen. In immunoblots, murine monoclonal and polyclonal human Bet v 1-specific Abs detected, besides a Bet v 1 monomer of 17 kDa, a dimer of 34 kDa. In dynamic light scattering, Bet v 1 appeared as dimers and even multimers, but a single condition could be defined where it behaved exclusively monomerically. Small-angle x-ray scattering of the monomeric and dimeric samples resulted in diagrams agreeing with the calculated models. Circular dichroism measurements indicated that the structure of Bet v 1 was preserved under monomeric conditions. Skin tests in Bet v 1-allergic mice were positive with Bet v 1 dimer, but remained negative using the monomer. Furthermore, in contrast to dimeric Bet v 1, the monomer was less capable of activating murine memory B cells for IgE production in vivo. Our data indicate that the presentation of two identical epitopes by dimerized allergens is a precondition for cross-linking of IgE on mast cells and B lymphocytes. PMID- 16272320 TI - Impaired assembly results in the accumulation of multiple HLA-C heavy chain folding intermediates. AB - Class I MHC H chains assemble with beta2-microglobulin (beta2m) and are loaded with peptide Ags through multiple folding steps. When free of beta2m, human H chains react with Abs to linear epitopes, such as L31. Immunodepletion and coimmunoprecipitation experiments, performed in this study, detected a preferential association of L31-reactive, beta2m-free H chains with calnexin in beta2m-defective cells, and with calreticulin and TAP in beta2m-expressing cells. In beta2m-defective cells, the accumulation of calnexin-bound H chains stoichiometrically exceeded their overall accumulation, a finding that supports both chaperoning preferences and distinct sorting abilities for different class I folds. No peptide species, in a mass range compatible with that of the classical class I ligands, could be detected by mass spectrometry of acidic eluates from L31-reactive HLA-Cw1 H chains. In vitro assembly experiments in TAP-defective T2 cells, and in cells expressing an intact Ag-processing machinery, demonstrated that L31 H chains are not only free of, but also unreceptive to, peptides. L31 and HC10, which bind nearly adjacent linear epitopes of the alpha1 domain alpha helix, reciprocally immunodepleted free HLA-C H chains, indicating the existence of a local un-/mis-folding involving the N-terminal end of the alpha1 domain alpha helix and peptide-anchoring residues of the class I H chain. Thus, unlike certain murine free H chains, L31-reactive H chains are not the immediate precursors of conformed class I molecules. A model inferring their precursor product relationships with other known class I intermediates is presented. PMID- 16272321 TI - The TCRalpha locus control region specifies thymic, but not peripheral, patterns of TCRalpha gene expression. AB - The molecular mechanisms ensuring the ordered expression of TCR genes are critical for proper T cell development. The mouse TCR alpha-chain gene locus contains a cis-acting locus control region (LCR) that has been shown to direct integration site-independent, lymphoid organ-specific expression of transgenes in vivo. However, the fine cell type specificity and developmental timing of TCRalpha LCR activity are both still unknown. To address these questions, we established a transgenic reporter model of TCRalpha LCR function that allows for analysis of LCR activity in individual cells by the use of flow cytometry. In this study we report the activation of TCRalpha LCR activity at the CD4-CD8-CD25 CD44- stage of thymocyte development that coincides with the onset of endogenous TCRalpha gene rearrangement and expression. Surprisingly, TCRalpha LCR activity appears to decrease in peripheral T cells where TCRalpha mRNA is normally up regulated. Furthermore, LCR-linked transgene activity is evident in gammadelta T cells and B cells. These data show that the LCR has all the elements required to reliably reproduce a developmentally correct TCRalpha-like expression pattern during thymic development and unexpectedly indicate that separate gene regulatory mechanisms are acting on the TCRalpha gene in peripheral T cells to ensure its high level and fine cell type-specific expression. PMID- 16272322 TI - The extent of histone acetylation correlates with the differential rearrangement frequency of individual VH genes in pro-B cells. AB - During B lymphocyte development, Ig heavy and L chain genes are assembled by V(D)J recombination. Individual V, D, and J genes rearrange at very different frequencies in vivo, and the natural variation in recombination signal sequence does not account for all of these differences. Because a permissive chromatin structure is necessary for the accessibility of VH genes for VH to DJH recombination, we hypothesized that gene rearrangement frequency might be influenced by the extent of histone modifications. Indeed, we found in freshly isolated pro-B cells from muMT mice a positive correlation between the level of enrichment of VHS107 genes in the acetylated histone fractions as assayed by chromatin immunoprecipitation, and their relative rearrangement frequency in vivo. In the VH7183 family, the very frequently rearranging VH81X gene showed the highest association with acetylated histones, especially in the newborn. Together, our data show that the extent of histone modifications in pro-B cells should be considered as a mechanism by which accessibility and the rearrangement level of individual VH genes is regulated. PMID- 16272323 TI - Up-regulation of CC chemokine ligand 20 expression in human airway epithelium by IL-17 through a JAK-independent but MEK/NF-kappaB-dependent signaling pathway. AB - CCL20, like human beta-defensin (hBD)-2, is a potent chemoattractant for CCR6 positive immature dendritic cells and T cells in addition to recently found antimicrobial activities. We previously demonstrated that IL-17 is the most potent cytokine to induce an apical secretion and expression of hBD-2 by human airway epithelial cells, and the induction is JAK/NF-kappaB-dependent. Similar to hBD-2, IL-17 also induced CCL20 expression, but the nature of the induction has not been elucidated. Compared with a panel of cytokines (IL-1alpha, 1beta, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 15, 16, 18, IFN-gamma, GM-CSF, and TNF-alpha), IL-17 was as potent as IL-1alpha, 1beta, and TNF-alpha, with a time- and dose dependent phenomenon in stimulating CCL20 expression in both well-differentiated primary human and mouse airway epithelial cell culture systems. The stimulation was largely dependent on the treatment of polarized epithelial cultures from the basolateral side with IL-17, achieving an estimated 4- to 10-fold stimulation at both message and protein levels. More than 90% of induced CCL20 secretion was toward the basolateral compartment (23.02 +/- 1.11 ng/chamber/day/basolateral vs 1.82 +/- 0.82 ng/chamber/day/apical). Actinomycin D experiments revealed that enhanced expression did not occur at mRNA stability. Inhibitor studies showed that enhanced expression was insensitive to inhibitors of JAK/STAT, p38, JNK, and PI3K signaling pathways, but sensitive to inhibitors of MEK1/2 and NF-kappaB activation, suggesting a MEK/NF-kappaB-based mechanism. These results suggest that IL-17 can coordinately up-regulate both hBD-2 and CCL20 expressions in airways through differentially JAK-dependent and -independent activations of NF kappaB-based transcriptional mechanisms, respectively. PMID- 16272324 TI - Identification of soluble N-ethylmaleimide-sensitive factor attachment protein receptor exocytotic machinery in human plasma cells: SNAP-23 is essential for antibody secretion. AB - Plasma cells (PC) are B-lymphocytes terminally differentiated in a postmitotic state, with the unique purpose of manufacturing and exporting Igs. Despite the importance of this process in the survival of vertebrates, no studies have been made to understand the molecular events that regulate Ig exocytosis by PC. The present study explores the possible presence of the soluble N-ethylmaleimide sensitive factor attachment protein receptor (SNARE) system in human PC, and examines its functional role in Ig secretion. Syntaxin-2, Syntaxin-3, Syntaxin-4, vesicle-associated membrane protein (VAMP)-2, VAMP-3, and synaptosome-associated protein (SNAP)-23 could be readily detected in normal human PC obtained from intestinal lamina propria and blood, as well as in human PC lines. Because SNAP 23 plays a central role in SNAREs complex formation, it was chosen to examine possible functional implications of the SNARE system in PC Ig secretion. When recombinant SNAP-23 fusion protein was introduced into the cells, a complete abolishment of Ig production was observed in the culture supernatants of PC lines, as well as in those of normal PC. These results provide insights, for the first time, into the molecular machinery of constitutive vesicular trafficking in human PC Ig secretion and present evidence indicating that at least SNAP-23 is essential for Ab production. PMID- 16272325 TI - Structural requirements for the interaction of human IgA with the human polymeric Ig receptor. AB - Transport of polymeric IgA onto mucosal surfaces to become secretory IgA is mediated by the polymeric Ig receptor (pIgR). To study the interaction of human dimeric IgA (dIgA) (the predominant form of IgA polymer) with the human pIgR (hpIgR), we generated recombinant wild-type dIgA1 and dIgA2m(1) and various mutant dIgA1 and analyzed their interaction with a recombinant human secretory component and membrane-expressed hpIgR. We found that wild-type dIgA1 and dIgA2m(1) bound to recombinant human secretory component with similar affinity and were transcytosed by the hpIgR to the same extent. Mutation of the IgA Calpha2 domain residue Cys311 to Ser reduced binding to hpIgR, possibly through disruption of noncovalent interactions between the Calpha2 domain and domain 5 of the receptor. Within the Calpha3 domain of IgA1, we found that combined mutation of residues Phe411, Val413, and Thr414, which lie close to residues previously implicated in hpIgR binding, abolished interaction with the receptor. Mutation of residue Lys377, located very close to this same region, perturbed receptor interaction. In addition, 4 aa (Pro440-Phe443), which lie on a loop at the domain interface and form part of the binding site for human FcalphaRI, appear to contribute to hpIgR binding. Lastly, use of a monomeric IgA1 mutant lacking the tailpiece revealed that the tailpiece does not occlude hpIgR-binding residues in IgA1 monomers. This directed mutagenesis approach has thus identified motifs lying principally across the upper surface of the Calpha3 domain (i.e., that closest to Calpha2) critical for human pIgR binding and transcytosis. PMID- 16272326 TI - The MHC of the duck (Anas platyrhynchos) contains five differentially expressed class I genes. AB - MHC class I proteins mediate a variety of functions in antiviral defense. In humans and mice, three MHC class I loci each contribute one or two alleles and each can present a wide variety of peptide Ags. In contrast, many lower vertebrates appear to use a single MHC class I locus. Previously we showed that a single locus was predominantly expressed in the mallard duck (Anas platyrhynchos) and that locus was adjacent to the polymorphic transporter for the Ag-processing (TAP2) gene. Characterization of a genomic clone from the same duck now allows us to compare genes to account for their differential expression. The clone carried five MHC class I genes and the TAP genes in the following gene order: TAP1, TAP2, UAA, UBA, UCA, UDA, and UEA. We designated the predominantly expressed gene UAA. Transcripts corresponding to the UDA locus were expressed at a low level. No transcripts were found for three loci, UBA, UCA, and UEA. UBA had a deletion within the promoter sequences. UCA carried a stop codon in-frame. UEA did not have a polyadenylation signal sequence. All sequences differed primarily in peptide-binding pockets and otherwise had the hallmarks of classical MHC class I alleles. Despite the presence of additional genes in the genome, the duck expresses predominantly one MHC class I gene. The limitation to one expressed MHC class I gene may have functional consequences for the ability of ducks to eliminate viral pathogens, such as influenza. PMID- 16272327 TI - The characterization of intraepithelial lymphocytes, lamina propria leukocytes, and isolated lymphoid follicles in the large intestine of mice infected with the intestinal nematode parasite Trichuris muris. AB - Despite a growing understanding of the role of cytokines in immunity to the parasitic helminth Trichuris muris, the local effector mechanism culminating in the expulsion of worms from the large intestine is not known. We used flow cytometry and immunohistochemistry to characterize the phenotype of large intestinal intraepithelial lymphocytes (IEL) and lamina propria leukocytes (LPL) from resistant and susceptible strains of mouse infected with T. muris. Leukocytes accumulated in the epithelium and lamina propria after infection, revealing marked differences between the different strains of mouse. In resistant mice, which mount a Th2 response, the number of infiltrating CD4+, CD8+, B220+, and F4/80+ IEL and LPL was generally highest around the time of worm expulsion from the gut, at which point the inflammation was dominated by CD4+ IEL and F4/80+ LPL. In contrast, in susceptible mice, which mount a Th1 response, the number of IEL and LPL increased more gradually and was highest after a chronic infection had developed. At this point, CD8+ IEL and F4/80+ LPL were predominant. Therefore, this study reveals the local immune responses underlying the expulsion of worms or the persistence of a chronic infection in resistant and susceptible strains of mouse, respectively. In addition, for the first time, we illustrate isolated lymphoid follicles in the large intestine, consisting of B cells interspersed with CD4+ T cells and having a central zone of rapidly proliferating cells. Furthermore, we demonstrate the organogenesis of these structures in response to T. muris infection. PMID- 16272329 TI - A live diarrheal vaccine imprints a Th2 cell bias and acts as an anti inflammatory vaccine. AB - An experimental vaccine for enterotoxigenic Escherichia coli (ETEC) composed of a live, attenuated Salmonella vector-expressing enterotoxigenic E. coli fimbriae, colonization factor Ag I (CFA/I), stimulated a biphasic Th cell response when given orally and suppressed the normally produced proinflammatory response. Such suppression was also evident upon the Salmonella-CFA/I infection of macrophages resulting in diminished TNF-alpha, IL-1, and IL-6 production and suggesting that the CFA/I fimbrial expression by Salmonella may protect against a proinflammatory disease. To test this hypothesis, SJL/J mice were vaccinated with Salmonella CFA/I construct 1 or 4 wk before induction of experimental autoimmune encephalomyelitis using an encephalitogenic proteolipid protein peptide, PLP(139 151). Mice receiving Salmonella-CFA/I vaccine recovered completely from mild acute clinical disease and showed only mild inflammatory infiltrates in the spinal cord white and gray matter. This protective effect was accompanied by a loss of encephalitogenic IFN-gamma-secreting Th cells and was replaced with an increase in IL-4, IL-10, and IL-13 secretion. Collectively, these data suggested that Salmonella-CFA/I is an anti-inflammatory vaccine that down-regulates proinflammatory cells and confers protection against a proinflammatory disease, experimental autoimmune encephalomyelitis, via immune deviation. PMID- 16272330 TI - Combined effects of ATP on the therapeutic efficacy of antimicrobial drug regimens against Mycobacterium avium complex infection in mice and roles of cytosolic phospholipase A2-dependent mechanisms in the ATP-mediated potentiation of antimycobacterial host resistance. AB - ATP, which serves as a mediator of intramacrophage signaling pathways through purinoceptors, is known to potentiate macrophage antimycobacterial activity. In this study we examined the effects of ATP in potentiating host resistance to Mycobacterium avium complex (MAC) infection in mice undergoing treatment with a drug regimen using clarithromycin and rifamycin and obtained the following findings. First, the administration of ATP in combination with the clarithromycin and rifamycin regimen accelerated bacterial elimination in MAC-infected mice without causing changes in the histopathological features or the mRNA expression of pro- or anti-inflammatory cytokines from those in the mice not given ATP. Second, ATP potentiated the anti-MAC bactericidal activity of macrophages cultivated in the presence of clarithromycin and rifamycin. This effect of ATP was closely related to intracellular Ca2+ mobilization and was specifically blocked by a cytosolic phospholipase A2 (cPLA2) inhibitor, arachidonyl trifluoromethylketone. Third, intramacrophage translocation of membranous arachidonic acid molecules to MAC-containing phagosomes was also specifically blocked by arachidonyl trifluoromethylketone. In the confocal microscopic observation of MAC-infected macrophages, ATP enhanced the intracellular translocation of cPLA2 into MAC-containing phagosomes. These findings suggest that ATP increases the host anti-MAC resistance by potentiating the antimycobacterial activity of host macrophages and that the cPLA2-dependent generation of arachidonic acid from the phagosomal membrane is essential for such a phenomenon. PMID- 16272328 TI - MyD88-dependent and -independent murine cytomegalovirus sensing for IFN-alpha release and initiation of immune responses in vivo. AB - Antiviral immunity requires early and late mechanisms in which IFN-alpha and IL 12 play major roles. However, the initial events leading to their production remain largely unclear. Given the crucial role of TLR in innate recognition, we investigated their role in antiviral immunity in vivo. Upon murine CMV (MCMV) infection, both MyD88-/- and TLR9-/- mice were more susceptible and presented increased viral loads compared with C57BL/6, TLR2-/-, TLR3-/-, or TLR4-/- mice. However, in terms of resistance to infection, IFN-alpha production and in many other parameters of early inflammatory responses, the MyD88-/- mice showed a more defective response than TLR9-/- mice. In the absence of the TLR9/MyD88 signaling pathway, cytokine production was dramatically impaired with a complete abolition of bioactive IL-12p70 serum release contrasting with a high flexibility for IFN alpha release, which is initially (36 h) plasmacytoid dendritic cell- and MyD88 dependent, and subsequently (44 h) PDC-, MyD88-independent and, most likely, TLR independent. NK cells from MCMV-infected MyD88-/- and TLR9-/- mice displayed a severely impaired IFN-gamma production, yet retained enhanced cytotoxic activity. In addition, dendritic cell activation and critical inflammatory cell trafficking toward the liver were still effective. In the long term, except for isotype switching to MCMV-specific IgG1, the establishment of Ab responses was not significantly altered. Thus, our results demonstrate a critical requirement of TLR9 in the process of MCMV sensing to assure rapid antiviral responses, coordinated with other TLR-dependent and -independent events that are sufficient to establish adaptive immunity. PMID- 16272331 TI - Early interaction of Yersinia pestis with APCs in the lung. AB - Despite the importance of pneumonic plague, little is known of the early pulmonary immune responses that occur following inhalation of Yersinia pestis. Therefore, we conducted studies to identify the early target cells for uptake of Y. pestis in the lungs following intratracheal or i.v. inoculation. Following intratracheal inoculation, Y. pestis was rapidly internalized primarily by a distinctive population of CD11c+DEC-205+CD11b- cells in the airways, whereas i.v. inoculation resulted in uptake primarily by CD11b+CD11c- macrophages and granulocytes in lung tissues. The airway cells internalized and were infected by Y. pestis, but did not support active replication of the organism. Intratracheal inoculation of Y. pestis resulted in rapid activation of airway CD11c+ cells, followed within 24 h by the selective disappearance of these cells from the airways and lungs and the accumulation of apoptotic CD11c+ cells in draining lymph nodes. When CD11c+ cells in the airways were depleted using liposomal clodronate before infection, this resulted in a significantly increased replication of Y. pestis in the lungs and dissemination to the spleen and draining lymph nodes. These findings suggest that CD11c+ cells in the airways play an important role in suppressing the initial replication and dissemination of inhaled Y. pestis, although these results will also require confirmation using fully virulent strains of Y. pestis. Depletion of these airway cells by Y. pestis may therefore be one strategy the organism uses to overcome pulmonary defenses following inhalation of the organism. PMID- 16272332 TI - A dual role for TGF-beta1 in the control and persistence of fungal pneumonia. AB - TGF-beta1 (TGF) has been implicated in the pathogenesis of several chronic infections and is thought to promote microbial persistence by interfering with macrophage function. In rats with experimental pulmonary cryptococcosis, increased lung levels of TGF were present at 12 mo of infection. Within the lung, expression of TGF localized to epithelioid cells and foamy macrophages in areas of inflammation. Increased TGF expression was also observed in the lungs of experimentally infected mice and a patient with pulmonary cryptococcosis. TGF reduced Ab and serum-mediated phagocytosis of Cryptococcus neoformans by rat alveolar macrophages (AM) and peripheral blood monocytes, and this was associated with decreased chemokine production and oxidative burst. Interestingly, TGF treated rat AM limited both intracellular and extracellular growth of C. neoformans. Control of C. neoformans growth by TGF-treated rat AM was due to increased secretion of lysozyme, a protein with potent antifungal activity. The effects of TGF on the course of infection were dependent on the timing of TGF administration relative to the time of infection. TGF treatment of chronically infected rats resulted in reduced lung fungal burden, while treatment early in the course of infection resulted in increased fungal burden. In summary, our studies suggest a dual role for TGF in persistent fungal pneumonia whereby it contributes to the local control of infection by enhancing macrophage antifungal efficacy through increased lysozyme secretion, while limiting inflammation by inhibiting macrophage/monocyte phagocytosis and reducing associated chemokine production and oxidative burst. PMID- 16272333 TI - Absence of thrombin-activatable fibrinolysis inhibitor protects against sepsis induced liver injury in mice. AB - Thrombin-activatable fibrinolysis inhibitor (TAFI), also known as carboxypeptidase R, has been implicated as an important negative regulator of the fibrinolytic system. In addition, TAFI is able to inactivate inflammatory peptides such as complement factors C3a and C5a. To determine the role of TAFI in the hemostatic and innate immune response to abdominal sepsis, TAFI gene deficient (TAFI-/-) and normal wild-type mice received an i.p. injection with Escherichia coli. Liver TAFI mRNA and TAFI protein concentrations increased during sepsis. In contrast to the presumptive role of TAFI as a natural inhibitor of fibrinolysis, TAFI-/- mice did not show any difference in E. coli-induced activation of coagulation or fibrinolysis, as measured by plasma levels of thrombin-anti-thrombin complexes and D-dimer and the extent of fibrin depositions in lung and liver tissues. However, TAFI-/- mice were protected from liver necrosis as indicated by histopathology and clinical chemistry. Furthermore, TAFI /- mice displayed an altered immune response to sepsis, as indicated by an increased neutrophil recruitment to the peritoneal cavity and a transiently increased bacterial outgrowth together with higher plasma TNF-alpha and IL-6 levels. These data argue against an important part for TAFI in the regulation of the procoagulant-fibrinolytic balance in sepsis and reveals a thus far unknown role of TAFI in the occurrence of hepatic necrosis. PMID- 16272334 TI - Antibody repertoire development in fetal and neonatal piglets. IX. Three pathogen associated molecular patterns act synergistically to allow germfree piglets to respond to type 2 thymus-independent and thymus-dependent antigens. AB - Newborn piglets maintained germfree (GF) cannot respond to either thymus dependent (TD) or type 2 thymus-independent Ags (TI-2) unless colonized with bacteria. We show here that pathogen-associated molecular patterns (PAMPs), including muramyl dipeptide (MDP), LPS, and a B-class CpG oligonucleotide (CpG B), can substitute for gut flora in the induction of neonatal immunoresponsiveness. These PAMPs alone or in combination had little effect on serum IgG and IgA levels, but CpG-B and CpG-B + MDP elevated total IgM levels 3- to 7-fold above that seen in colonized controls after booster immunization. Although only CpG-B could alone stimulate immunoresponsiveness, co-administration of LPS or MDP resulted in a 5-fold increase in the IgG response to both immunogens. Co-administered MDP did not promote secondary IgG responses to either Ag but instead pronounced secondary IgM responses to the epitopes of both immunogens. LPS co-administered with CpG-B may promote class switch recombination or cause differentiation of previously switched cells that become responsive after exposure to CpG-B. Primary and secondary IgG responses equally recognized the epitopes of the TI-2 and TD immunogens, whereas IgM responses favored the TI 2 epitope. Because PAMPs alone can result in Abs to 2,4,6-triitrophenyl and FLU without immunization, it suggests they alone cause differentiation of B cells of the preimmune repertoire. The finding that both bacterial PAMPs and colonization are capable of stimulating Ab responses in both immunized and nonimmunized piglets suggests that PAMPs derived from host flora may play a major role in awakening adaptive immunity in neonates. PMID- 16272335 TI - In vivo protective role of human group IIa phospholipase A2 against experimental anthrax. AB - Anthrax is an acute disease caused by Bacillus anthracis. Some animal species are relatively resistant to anthrax infection. This trait has been correlated to the extent of the local inflammatory reaction, suggesting innate immunity to be the first line of defense against B. anthracis infection in nonimmunized hosts. Group IIA secreted phospholipase A2 (sPLA2-IIA) is produced in particular by macrophages and possesses potent antibacterial activity especially against Gram positive bacteria. We have previously shown in vitro that sPLA2-IIA kills both germinated B. anthracis spores and encapsulated bacilli. Here we show that sPLA2 IIA plays in vivo a protective role against experimental anthrax. Transgenic mice expressing human sPLA2-IIA are resistant to B. anthracis infection. In addition, in vivo administration of recombinant human sPLA2-IIA protects mice against B. anthracis infection. The protective effect was observed both with a highly virulent encapsulated nontoxinogenic strain and a wild-type encapsulated toxinogenic strain, showing that toxemia did not hinder the sPLA2-IIA-afforded protection. sPLA2-IIA, a natural component of the immune system, may thus be considered a novel therapeutic agent to be used in adjunct with current therapy for treating anthrax. Its anthracidal activity would be effective even against strains resistant to multiple antibiotics. PMID- 16272336 TI - Francisella tularensis induces aberrant activation of pulmonary dendritic cells. AB - Francisella tularensis is an obligate intracellular bacterium that induces severe, acute, often fatal disease when acquired by the respiratory route. Despite the seriousness of this pathogen, very little is understood about its interaction with key target cells in the airways and lungs (alveolar macrophages and airway dendritic cells (DC)) after inhalation. In this study we demonstrate replication of F. tularensis in primary DC. Early after infection, F. tularensis induced increased expression of MHC class II and CD86 on DC, but not macrophages. This was followed by depletion of DC from the airways and lungs. Despite logarithmic replication and phenotypic maturation of DC, F. tularensis failed to induce production of several key proinflammatory cytokines, including TNF-alpha and IL-6, from DC. However, F. tularensis infection did elicit production of the potent immunosuppressive cytokine, TGF-beta. Furthermore, F. tularensis actively suppressed the ability of DC to secrete cytokines in response to specific TLR agonists. Finally, we also found that infection of DC and macrophages in the lungs appears to actually increase the severity of pulmonary infection with F. tularensis. For example, depletion of airway DC and alveolar macrophages before infection resulted in significantly prolonged survival times. Together, these data suggest F. tularensis is able to selectively uncouple Ag-presenting functions from proinflammatory cytokine secretion by critical APCs in the lungs, which may serve to create a relatively immunosuppressive environment favorable to replication and dissemination of the organism. PMID- 16272338 TI - Optimization of CD4+ T lymphocyte response to human cytomegalovirus nuclear IE1 protein through modifications of both size and cellular localization. AB - We have previously reported that the CD4+ T lymphocyte response against nuclear human CMV IE1 protein depends in part on endogenous MHC class II presentation. To optimize presentation by HLA-DR of the nuclear IE1 protein and increase the response by CD4+ T cells, we have constructed two different adenovirus vectors containing mutant versions of IE1, containing a HLA-DR3 epitope, fused to GFP. The first construct consisted of a sequence of 46 aa encoded by exon 4, called GFP-IE1 (86-131). The second construct consisted of the whole IE1 mutated on exon 4 nuclear localization signals, identified in this study, and deleted of already known exon 2 nuclear localization signals (GFP-IE1M). Both of these IE1 vectors expressed proteins with cytoplasmic localization, as evidenced by GFP expression, as opposed to control GFP-IE1, which was nuclear. GFP-IE1 (86-131) induced IE1 specific CD4+ T cell clone response that was >30-fold more potent than that against GFP-IE1 and GFP-IE1M. The CD4+ T cell response was due to endogenous presentation followed by exogenous presentation at later time points. Presentation was dependent on both proteasome and acidic compartments. GFP-IE1 (86-131) was rapidly degraded by the APC, which may account for better presentation. Our data show potentiation of the CD4+ T cell response to a specific epitope through shortening and relocation of an otherwise nuclear protein and suggest applications in vaccination. PMID- 16272337 TI - Neurokinin 1 receptor signaling affects the local innate immune defense against genital herpes virus infection. AB - We show that genital infection with neurotropic HSV type 2 (HSV-2) induced a significant increase of the neuropeptide substance P (SP) within the genital tract of mice. SP was shown to weakly interfere with the HSV-2 replication. Furthermore, lack of SP signaling through the use of mice deficient in the SP receptor, neurokinin 1 receptor (NK1R), revealed an important role for SP in the innate defense against HSV-2. NK1R-deficient mice had significantly enhanced levels of HSV-2 in the genital tract and in the CNS following infection and a significantly accelerated disease progression, which was associated with an impaired NK cell activity locally in the vagina. Lack of NK1R signaling did, however, not impair the animals' ability to mount a protective immune response to HSV-2 following vaccination with an attenuated virus. Both NK1R+/+ and NK1R-/- mice developed strong HSV-2-specific Th1 T cell responses following vaccination. No genital viral replication was observed in either vaccinated NK1R-deficient or NK1R+/+ control animals following a genital HSV-2 challenge, and all of these animals survived without any symptoms of disease. In conclusion, the present results indicate that SP and NK1R signaling contributes to the innate resistance against HSV-2 infection in mice. PMID- 16272339 TI - Requisite H2k role in NK cell-mediated resistance in acute murine cytomegalovirus infected MA/My mice. AB - Human CMV infections are a major health risk in patients with dysfunctional or compromised immunity, especially in patients with NK cell deficiencies, as these are frequently associated with high morbidity and mortality. In experimental murine CMV (MCMV) infections, Ly49H activation receptors on C57BL/6 (B6) NK cells engage m157 viral ligands on MCMV-infected cells and initiate dominant virus control. In this study, we report that MCMV resistance in MA/My relies on Ly49H independent NK cell-mediated control of MCMV infection as NK cells in these mice do not bind anti-Ly49H mAb or soluble m157 viral ligands. We genetically compared MA/My resistance with MCMV susceptibility in genealogically and NK gene complex Ly49 haplotype-related C57L mice. We found that MCMV resistance strongly associated with polymorphic H2k-linked genes, including MHC and non-MHC locations by analysis of backcross and intercross progeny. The H2b haplotype most frequently, but not absolutely, correlated with MCMV susceptibility, thus confirming a role for non-MHC genes in MCMV control. We also demonstrate a definite role for NK cells in H2k-type MCMV resistance because their removal from C57L.M-H2k mice before MCMV infection diminished immunity. NK gene complex-linked polymorphisms, however, did not significantly influence MCMV control. Taken together, effective NK cell-mediated MCMV control in this genetic system required polymorphic H2k genes without need of Ly49H-m157 interactions. PMID- 16272340 TI - Antibodies and CD8+ T cells are complementary and essential for natural resistance to a highly lethal cytopathic virus. AB - It is believed that CD8+ T lymphocytes or Abs can independently clear many primary viral infections, including those caused by Orthopoxviruses (OPV), a genus that includes the human pathogens variola and monkeypox and the vaccine species vaccinia virus. However, most experiments addressing the role of Abs and CD8+ T cells in protection have used viruses that are not specific for the host. In the present study, we used the mouse-specific OPV ectromelia virus and mice deficient in CD40, B cells, or CD8+ T cells and adoptive transfers of CD8+ T or B lymphocytes to show that the protection afforded by CD8+ T cells is incomplete. Despite sustained CD8+ T cell responses, in the absence of Ab responses ectromelia virus persists. This results in delayed disease and inexorably leads to death. Therefore, CD8+ T lymphocytes and Abs are not redundant but complementary and essential to survive infections with a highly pathogenic viruses in the natural host. PMID- 16272341 TI - Chronic ethanol ingestion in rats decreases granulocyte-macrophage colony stimulating factor receptor expression and downstream signaling in the alveolar macrophage. AB - Although it is well recognized that alcohol abuse impairs alveolar macrophage immune function and renders patients susceptible to pneumonia, the mechanisms are incompletely understood. Alveolar macrophage maturation and function requires priming by GM-CSF, which is produced and secreted into the alveolar space by the alveolar epithelium. In this study, we determined that although chronic ethanol ingestion (6 wk) in rats had no effect on GM-CSF expression within the alveolar space, it significantly decreased membrane expression of the GM-CSF receptor in alveolar macrophages. In parallel, ethanol ingestion decreased cellular expression and nuclear binding of PU.1, the master transcription factor that activates GM-CSF-dependent macrophage functions. Furthermore, treatment of ethanol-fed rats in vivo with rGM-CSF via the upper airway restored GM-CSF receptor membrane expression as well as PU.1 protein expression and nuclear binding in alveolar macrophages. Importantly, GM-CSF treatment also restored alveolar macrophage function in ethanol-fed rats, as reflected by endotoxin stimulated release of TNF-alpha and bacterial phagocytosis. We conclude that ethanol ingestion dampens alveolar macrophage immune function by decreasing GM CSF receptor expression and downstream PU.1 nuclear binding and that these chronic defects can be reversed relatively quickly with rGM-CSF treatment in vivo. PMID- 16272342 TI - Decoupling of carbohydrate binding and MASP-2 autoactivation in variant mannose binding lectins associated with immunodeficiency. AB - Mannan-binding lectin (MBL) initiates complement activation by binding to arrays of carbohydrates on the surfaces of pathogenic microorganisms and activating MBL associated serine proteases (MASPs). Separate point mutations to the collagenous domain of human MBL are associated with immunodeficiency, caused by reduced complement activation by the variant MBLs as well as by lower serum MBL concentrations. In the work reported here, we have used the well characterized rat lectin pathway to analyze the molecular and functional defects associated with two of the variant proteins. Mutations Gly25 --> Asp and Gly28 --> Glu create comparable structural changes in rat MBL but the G28E variant activates complement >10-fold less efficiently than the G25D variant, which in turn has approximately 7-fold lower activity than wild-type MBL. Analysis of mutant MBL . MASP-2 complexes assembled from recombinant components shows that reduced complement activation by both mutant MBLs is caused by failure to activate MASP-2 efficiently on binding to a mannan-coated surface. Disruption of MBL-MASP-2 interactions as well as to changes in oligomeric structure and reduced binding to carbohydrate ligands compared with wild-type MBL probably account for the intermediate phenotype of the G25D variant. However, carbohydrate binding and MASP-2 activation are ostensibly completely decoupled in complexes assembled from the G28E mutant, such that the rate of MASP-2 activation is no greater than the basal rate of zymogen MASP-2 autoactivation. Analogous molecular defects in human MBL probably combine to create the mutant phenotypes of immunodeficient individuals. PMID- 16272343 TI - Increased susceptibility to apoptosis of CD56dimCD16+ NK cells induces the enrichment of IFN-gamma-producing CD56bright cells in tuberculous pleurisy. AB - Tuberculous pleuritis is a good model for the study of specific cells at the site of active Mycobacterium tuberculosis (Mtb) infection. We investigated the frequency and phenotype of NK cells in paired samples of peripheral blood and pleural fluid (PF) from patients with tuberculosis (TB) or parapneumonic infection. We demonstrated for the first time a reduction of NK cells in PF from TB with an enrichment in the CD56brightCD16- subset. In agreement, in PF NK cells we observed an increased expression of CD94, NKG2A, CD62L, and CCR7 molecules and lower expression of Bcl-2 and perforin. The activation markers CD69 and HLA-DR were also increased. The enrichment in the CD56bright subset was due to an increased susceptibility to apoptosis of CD56+CD16+ NK cells mediated by heat labile and stable soluble factors present in tuberculous effusions and not in PF from other etiologies. Furthermore, in TB patients, Mtb-induced IFN-gamma production by PF NK cells was not dependent on the presence of CD3+, CD19+, and CD14+ cells, suggesting a direct interaction of CD56bright cells with Mtb and/or the involvement of other accessory cells present at the site of Mtb infection. PMID- 16272344 TI - Both hemopoietic and resident cells are required for MyD88-dependent pulmonary inflammatory response to inhaled endotoxin. AB - Inhaled endotoxin induces an inflammatory response that contributes to the development and severity of asthma and other forms of airway disease. Here, we show that inhaled endotoxin-induced acute bronchoconstriction, TNF, IL-12p40, and KC production, protein leak, and neutrophil recruitment in the lung are abrogated in mice deficient for the adaptor molecule MyD88. Bronchoconstriction, inflammation, and protein leak are normal in Toll/IL-1R domain-containing adaptor inducing IFN-beta-deficient mice. MyD88 is involved in TLR, but also in IL-1R associated kinase 1-mediated IL-1R and -18R signaling. We exclude a role for IL-1 and IL-18 pathways in this response, as IL-1R1 and caspase-1 (ICE)-deficient mice develop lung inflammation while TLR4-deficient mice are unresponsive to inhaled LPS. Significantly, using bone marrow chimera, we demonstrate that both hemopoietic and resident cells are necessary for a full MyD88-dependent response to inhaled endotoxin; bronchoconstriction depends on resident cells while cytokine secretion is mediated by hemopoietic cells. PMID- 16272345 TI - The mechanism of superantigen-mediated toxic shock: not a simple Th1 cytokine storm. AB - The profound clinical consequences of Gram-positive toxic shock are hypothesized to stem from excessive Th1 responses to superantigens. We used a new superantigen sensitive transgenic model to explore the role of TCRalphabeta T cells in responses to staphylococcal enterotoxin B (SEB) in vitro and in two different in vivo models. The proliferative and cytokine responses of HLA-DR1 spleen cells were 100-fold more sensitive than controls and were entirely dependent on TCRalphabeta T cells. HLA-DR1 mice showed greater sensitivity in vivo to two doses of SEB with higher mortality and serum cytokines than controls. When d galactosamine was used as a sensitizing agent with a single dose of SEB, HLA-DR1 mice died of toxic shock whereas controls did not. In this sensitized model of toxic shock there was a biphasic release of cytokines, including TNF-alpha, at 2 h and before death at 7 h. In both models, mortality and cytokine release at both time points were dependent on TCRalphabeta T cells. Anti-TNF-alpha pretreatment was protective against shock whereas anti-IFN gamma pretreatment and delayed anti TNF-alpha treatment were not. Importantly, anti-TNF-alpha pretreatment inhibited the early TNF-alpha response but did not inhibit the later TNF-alpha burst, to which mortality has previously been attributed. Splenic T cells were shown definitively to be the major source of TNF-alpha during the acute cytokine response. Our results demonstrate unequivocally that TCRalphabeta T cells are critical for lethality in toxic shock but it is the early TNF-alpha response and not the later cytokine surge that mediates lethal shock. PMID- 16272346 TI - Contrasting effects of cyclooxygenase-1 (COX-1) and COX-2 deficiency on the host response to influenza A viral infection. AB - Influenza is a significant cause of morbidity and mortality worldwide despite extensive research and vaccine availability. The cyclooxygenase (COX) pathway is important in modulating immune responses and is also a major target of nonsteroidal anti-inflammatory drugs (NSAIDs) and the newer COX-2 inhibitors. The purpose of the present study was to examine the effect of deficiency of COX-1 or COX-2 on the host response to influenza. We used an influenza A viral infection model in wild type (WT), COX-1-/-, and COX-2-/- mice. Infection induced less severe illness in COX-2-/- mice in comparison to WT and COX-1-/- mice as evidenced by body weight and body temperature changes. Mortality was significantly reduced in COX-2-/- mice. COX-1-/- mice had enhanced inflammation and earlier appearance of proinflammatory cytokines in the BAL fluid, whereas the inflammatory and cytokine responses were blunted in COX-2-/- mice. However, lung viral titers were markedly elevated in COX-2-/- mice relative to WT and COX-1-/- mice on day 4 of infection. Levels of PGE2 were reduced in COX-1-/- airways whereas cysteinyl leukotrienes were elevated in COX-2-/- airways following infection. Thus, deficiency of COX-1 and COX-2 leads to contrasting effects in the host response to influenza infection, and these differences are associated with altered production of prostaglandins and leukotrienes following infection. COX-1 deficiency is detrimental whereas COX-2 deficiency is beneficial to the host during influenza viral infection. PMID- 16272347 TI - Positive and negative regulation of mast cell activation by Lyn via the FcepsilonRI. AB - Aggregation of the high affinity receptor for IgE (FcepsilonRI) induces activation of mast cells. In this study we show that upon low intensity stimulation of FcepsilonRI with monomeric IgE, IgE plus anti-IgE, or IgE plus low Ag, Lyn (a Src family kinase) positively regulates degranulation, cytokine production, and survival, whereas Lyn works as a negative regulator of high intensity stimulation with IgE plus high Ag. Low intensity stimulation suppressed Lyn kinase activity and its association with FcepsilonRI beta subunit, whereas high intensity stimulation enhanced Lyn activity and its association with FcepsilonRI beta. The latter induced much higher levels of FcepsilonRI beta phosphorylation and Syk activity than the former. Downstream positive signaling molecules, such as Akt and p38, were positively and negatively regulated by Lyn upon low and high intensity stimulations, respectively. In contrast, the negative regulators, SHIP and Src homology 2 domain-containing protein tyrosine phosphatase-1, interacted with FcepsilonRI beta, and their phosphorylation was controlled by Lyn. Therefore, we conclude that Lyn-mediated positive vs negative regulation depends on the intensity of the stimuli. Studies of mutant FcepsilonRI beta showed that FcepsilonRI beta subunit-ITAM (ITAM motif) regulates degranulation and cytokine production positively and negatively depending on the intensity of FcepsilonRI stimulation. Furthermore, Lyn-mediated negative regulation was shown to be exerted via the FcepsilonRI beta-ITAM. PMID- 16272349 TI - Abnormally differentiated subsets of intestinal macrophage play a key role in Th1 dominant chronic colitis through excess production of IL-12 and IL-23 in response to bacteria. AB - Disorders in enteric bacteria recognition by intestinal macrophages (Mphi) are strongly correlated with the pathogenesis of chronic colitis; however the precise mechanisms remain unclear. The aim of the current study was to elucidate the roles of Mphi in intestinal inflammation by using an IL-10-deficient (IL-10-/-) mouse colitis model. GM-CSF-induced bone marrow-derived Mphi (GM-Mphi) and M-CSF induced bone marrow-derived Mphi (M-Mphi) were generated from bone marrow CD11b+ cells. M-Mphi from IL-10-/- mice produced abnormally large amounts of IL-12 and IL-23 upon stimulation with heat-killed whole bacteria Ags, whereas M-Mphi from wild-type (WT) mice produced large amounts of IL-10 but not IL-12 or IL-23. In contrast, IL-12 production by GM-Mphi was not significantly different between WT and IL-10-/- mice. In ex vivo experiments, cytokine production ability of colonic lamina propria Mphi (CLPMphi) but not splenic Mphi from WT mice was similar to that of M-Mphi, and CLPMphi but not splenic Mphi from IL-10-/- mice also showed abnormal IL-12p70 hyperproduction upon stimulation with bacteria. Surprisingly, the abnormal IL-12p70 hyperproduction from M-Mphi from IL-10-/- mice was improved by IL-10 supplementation during the differentiation process. These results suggest that CLPMphi and M-Mphi act as anti-inflammatory Mphi and suppress excess inflammation induced by bacteria in WT mice. In IL-10-/- mice, however, such Mphi subsets differentiated into an abnormal phenotype under an IL-10-deficient environment, and bacteria recognition by abnormally differentiated subsets of intestinal Mphi may lead to Th1-dominant colitis via IL-12 and IL-23 hyperproduction. Our data provide new insights into the intestinal Mphi to gut flora relationship in the development of colitis in IL-10-/- mice. PMID- 16272348 TI - Involvement of TLR4/type I IL-1 receptor signaling in the induction of inflammatory mediators and cell death induced by ethanol in cultured astrocytes. AB - Activated astroglial cells are implicated in neuropathogenesis of many infectious and inflammatory diseases of the brain. A number of inflammatory mediators and cytokines have been proposed to play a key role in glial cell-related brain damage. Cytokine production seems to be initiated by signaling through TLR4/type I IL-1R (IL-1RI) in response to their ligands, LPS and IL-1beta, playing vital roles in innate host defense against infections, inflammation, injury, and stress. We have shown that glial cells are stimulated by ethanol, up-regulating cytokines and inflammatory mediators associated with TLR4 and IL-1RI signaling pathways in brain, suggesting that ethanol may contribute to brain damage via inflammation. We explore the possibility that ethanol, in the absence of LPS or IL-1beta, triggers signaling pathways and inflammatory mediators through TLR4 and/or IL-1RI activation in astrocytes. We show in this study that ethanol, at physiologically relevant concentrations, is capable of inducing rapid phosphorylation within 10 min of IL-1R-associated kinase, ERK1/2, stress activated protein kinase/JNK, and p38 MAPK in astrocytes. Then an activation of NF-kappaB and AP-1 occurs after 30 min of ethanol treatment along with an up regulation of inducible NO synthase and cyclooxygenase-2 expression. Finally, we note an increase in cell death after 3 h of treatment. Furthermore, by using either anti-TLR4- or anti-IL-1RI-neutralizing Abs, before and during ethanol treatment, we inhibit ethanol-induced signaling events, including NF-kappaB and AP-1 activation, inducible NO synthase, and cyclooxygenase-2 up-regulation and astrocyte death. In summary, these findings indicate that both TLR4 and IL-1RI activation occur upon ethanol treatment, and suggest that signaling through these receptors mediates ethanol-induced inflammatory events in astrocytes and brain. PMID- 16272350 TI - The role of complement in cryoglobulin-induced immune complex glomerulonephritis. AB - Many forms of glomerulonephritis are triggered by Ab localization in the glomerulus, but the mechanisms by which this induces glomerular inflammation are not fully understood. In this study we investigated the role of complement in a mouse model of cryoglobulin-induced immune complex glomerulonephritis. Several complement-deficient mice on a C57BL/6 and BALB/c genetic background were used and compared with strain-matched, wild-type controls. Cryoglobulinemia was induced by i.p. injection of 6-19 hybridoma cells producing an IgG3 cryoglobulin with rheumatoid factor activity against IgG2a of allotype a present in BALB/c, but not C57BL/6, mice. Thus, the cryoprecipitate in C57BL/6 mice consisted of the IgG3 cryoglobulin only (type I cryoglobulinemia) compared with IgG3-IgG2a complexes in BALB/c (type II cryoglobulinemia). The survival of mice was not affected by complement deficiency. Glomerular influx of neutrophils was significantly less in C3-, factor B-, and C5-deficient mice compared with wild type and C1q-deficient mice. It did not correlate with C3 deposition, but did correlate with the amount of C6 deposited. Deficiency of CD59a, the membrane inhibitor of the membrane attack complex, did not induce an increase in neutrophil infiltration, suggesting that the generation of C5a accounts for the effects observed. There was no apparent difference between cryoglobulinemia types I and II regarding the role of complement. Our results suggest that in this model of cryoglobulin-induced glomerulonephritis the neutrophil influx was mediated by C5 activation with the alternative pathway playing a prominent role in its cleavage. Thus, blocking C5 is a potential therapeutic strategy for preventing renal injury in cryoglobulinemia. PMID- 16272352 TI - Resolution of inflammation: prostaglandin E2 dissociates nuclear trafficking of individual NF-kappaB subunits (p65, p50) in stimulated rheumatoid synovial fibroblasts. AB - NF-kappaB transcription factors regulate inflammatory responses to cytokines such as IL-1beta and TNF-alpha. We tested whether PGE2 regulated nuclear localization of individual NF-kappaB subunits, p65 and p50, in synovial fibroblasts harvested from patients with rheumatoid arthritis (RA). IL-1beta/TNF-alpha stimulated the translocation of p65 and p50 from the cytosol to the nucleus of human RA synovial fibroblasts, as well as NF-kappaB activation measured by luciferase reporter assay. PGE2 (10 nM, 6 h) enhanced p50, but inhibited p65 translocation and NF kappaB activation. In contrast, depletion of endogenous PGE2 by ibuprofen (100 microM) and celecoxib (5 microM) enhanced p65, but inhibited p50 nuclear translocation as well as binding to NF-kappaB DNA binding sites. PGE2 also blocked IL-1beta/TNF-alpha-stimulated ERK activation, and the ERK inhibitor, PD98059, mimicked PGE2 in blocking p65, but enhancing p50 nuclear translocation, suggesting that the effects of PGE2 on p65 and p50 are mediated via effects on ERK. PGE2 also enhanced the expression of IkappaBalpha in an ERK-independent manner, suggesting that PGE2 inhibits NF-kappaB activation by both ERK-dependent and -independent mechanisms. Our data indicate that PGE2 may act to attenuate cytokine-induced inflammatory responses in RA synovial fibroblasts via regulation of the localization of specific NF-kappaB family dimers. PMID- 16272351 TI - Differentiation to the CCR2+ inflammatory phenotype in vivo is a constitutive, time-limited property of blood monocytes and is independent of local inflammatory mediators. AB - It is proposed that CCR2+ monocytes are specifically recruited to inflammatory sites, whereas CCR2- monocytes are recruited to normal tissue to become resident macrophages. Whether these subsets represent separate lineages, how differential trafficking is regulated and whether monocytes undergo further differentiation is uncertain. Using a mouse model of autoimmune uveoretinitis we examined monocyte trafficking to the inflamed retina in vivo. We show that bone marrow-derived CD11b+ F4/80- monocytes require 24 to 48 h within the circulation and lymphoid system before acquiring the CCR2+ phenotype and trafficking to the inflamed retina is enabled. This phenotype, and the capacity to traffic were lost by 72 h. Monocyte CCR2 expression followed a similar time course in normal mice indicating that differentiation to an inflammatory phenotype is a constitutive, time-limited property, independent of local inflammatory mediators. Phenotypic analysis of adoptively transferred cells indicated that circulating inflammatory monocytes also differentiate into CD11c+ and B220+ dendritic cells and F4/80+ tissue macrophages in vivo. Our data supports the hypothesis of continuous extravasation and progressive differentiation over time of inflammatory monocytes in the circulation rather than replication within the actively inflamed tissue, and supports the concept of myeloid dendritic cell differentiation from trafficking monocytes under physiological conditions in vivo. PMID- 16272353 TI - CXCR2/CXCR2 ligand biology during lung transplant ischemia-reperfusion injury. AB - Lung transplantation is a therapeutic option for a number of end-stage pulmonary disorders. Early lung allograft dysfunction (ischemia-reperfusion injury) continues to be the most common cause of early mortality after lung transplantation and a significant risk factor for the development of bronchiolitis obliterans syndrome. Ischemia-reperfusion injury is characterized histopathologically by lung edema and a neutrophil predominate leukocyte extravasation. The specific mechanism(s) that recruit leukocytes to the lung during post-lung transplantation ischemia-reperfusion injury have not been fully elucidated. Because the ELR+ CXC chemokines are potent neutrophil chemoattractants, we investigated their role during post-lung transplantation ischemic-reperfusion injury. We found elevated levels of multiple ELR+ CXC chemokines in human bronchoalveolar lavage fluid from patients with ischemia reperfusion injury. Proof of concept studies using a rat orthotopic lung transplantation model of "cold" ischemic-reperfusion injury demonstrated an increase in lung graft neutrophil sequestration and injury. In addition, lung expression of CXCL1, CXCL2/3, and their shared receptor CXCR2 paralleled lung neutrophil infiltration and injury. Importantly, inhibition of CXCR2/CXCR2 ligand interactions in vivo led to a marked reduction in lung neutrophil sequestration and graft injury. Taken together these experiments support the notion that increased expression of ELR+ CXC chemokines and their interaction with CXCR2 plays an important role in the pathogenesis of post-lung transplantation cold ischemia-reperfusion injury. PMID- 16272354 TI - Proliferation response of leukemic cells to CD70 ligation oscillates with recurrent remission and relapse in a low-grade lymphoma. AB - Interactions of the TNF-related cell surface ligand CD70 with its receptor CD27 provide a costimulatory signal in B and T cell activation. Functional CD70-CD27 interactions could contribute to lymphoma and leukemia progression. This possibility was studied using DNA microarrays on a unique case of low-grade lymphoma/leukemia characterized by recurrent cycles of acute leukemic phase alternating with spontaneous remission. Upon induction of the acute phase expression of CD70 and CD27 in the leukemic cells increased 38- and 25-fold, respectively. Coexpression of membrane CD70 and CD27 on the leukemic (CD5+CD19+) cells was maximal 2-3 days following initiation of the attack. Soluble CD27 in the patient's serum was elevated during remission and further increased in the attack. Functional tests showed that neither anti-CD70 nor anti-CD27 Abs affect the rate of apoptosis. However, the anti-CD70 Ab specifically enhanced proliferation of the remission phase leukemic cells, whereas proliferation of the acute-phase counterparts that express higher level of membrane CD70 was unaffected. Hence, in this lymphoma/leukemia, membrane CD70 is presented on the leukemic cells in a responsive state during the remission and a nonresponsive state during the attack. Presumably, CD70 in its responsive state provides a costimulatory receptor for initiating the next acute phase while its nonresponsive state enables the remission. PMID- 16272355 TI - Expansion and contraction of HIV-specific CD4 T cells with short bursts of viremia, but physical loss of the majority of these cells with sustained viral replication. AB - Chronic infection with the HIV results in poor HIV-specific CD4 T cell proliferation, but more recent analyses using intracellular cytokine staining demonstrated that IFN-gamma-producing, HIV-specific CD4 T cells can be detected for years in HIV-infected subjects. Because it is not known whether the majority of HIV-specific T cells are lost or become dysfunctional, we examined the kinetics of the T cell response over an extended period of time using a panel of 10 HLA-DR tetramers loaded with HIV p24 peptides. Tetramer+ CD4 T cells were present at a relatively high frequency during acute infection, but the size of these populations substantially contracted following suppression of viral replication. Short-term cessation of antiretroviral therapy resulted in a burst of viral replication and concomitant expansion of tetramer+ CD4 T cells, and these populations again contracted following reinitiation of therapy. The kinetics with which these cell populations contracted were characteristic of effector T cells, a conclusion that was supported by their phenotypic (CCR7 CD45RA-) and functional properties (IFN-gamma+). Continued high-level viremia resulted in the physical loss of the majority of tetramer+ CD4 T cells, and the decline of HIV p24-specific CD4 T cells occurred more rapidly and was more substantial than the reduction of total CD4 T cell numbers. We conclude that the population of HIV p24-specific CD4 T cells is initially responsive to changes in the levels of viral Ags, but that the majority of these cells are lost in a setting of chronic viremia. PMID- 16272356 TI - A mimic of p21WAF1/CIP1 ameliorates murine lupus. AB - Systemic lupus erythematosus (SLE) is a progressive autoimmune disease characterized by the production of high levels of affinity-matured IgG autoantibodies to dsDNA and, possibly, visceral involvement. Pathogenic autoantibodies result from the activation and proliferation of autoreactive T and B lymphocytes stimulated by epitopes borne by nucleosomal histones. To inhibit the proliferation of autoreactive cells and abrogate the development of SLE, a novel tool, cell cycle inhibiting peptide therapy, was used. Thus, a peptidyl mimic of p21WAF1/CIP1 that inhibits the interaction between cyclin-dependent kinase 4 and type D cyclins abrogated the in vitro proliferative response of T cells to histones and T-independent and T-dependent proliferative responses of B cells. The WAF1/CIP1 mimic also abrogated the in vitro production of total and anti-dsDNA IgG Abs by B cells. Similarly, the p21WAF1/CIP1 construct inhibited the ex vivo T and B cell proliferative responses to histones and decreased the numbers of activated/memory B and T spleen cells. The alterations in the balance of spleen cell subsets resulted from proapoptotic effects of the p21WAF1/CIP1)construct on activated splenocytes. Finally, in vivo, four i.v. injections of the p21WAF1/CIP1 mimic were sufficient to inhibit the progression of the lupus-like syndrome in (NZB x NZW)F1 mice. The levels of anti-dsDNA IgG autoantibodies and the incidence and severity of renal involvement were lower in treated mice than in nontreated mice. Those observations open new avenues for the treatment of SLE and prompt us to evaluate the potential interest of peptidic therapy in human SLE. PMID- 16272357 TI - Transcription factor Nrf2 plays a pivotal role in protection against elastase induced pulmonary inflammation and emphysema. AB - Emphysema is one of the major pathological abnormalities associated with chronic obstructive pulmonary disease. The protease/antiprotease imbalance and inflammation resulting from oxidative stress have been attributed to the pathogenesis of emphysema. Nrf2 is believed to protect against oxidative tissue damage through the transcriptional activation of a battery of antioxidant enzymes. In this study, we investigated the protective role of Nrf2 in the development of emphysema using elastase-induced emphysema as our model system. We found that elastase-provoked emphysema was markedly exacerbated in Nrf2-knockout (KO) mice compared with wild-type mice. The severity of emphysema in Nrf2-KO mice correlated intimately with the degree of lung inflammation in the initial stage of elastase treatment. The highly inducible expression of antioxidant and antiprotease genes observed in wild-type alveolar macrophages was significantly attenuated in the lungs of Nrf2-KO mice. Interestingly, transplantation of wild type bone marrow cells into Nrf2-KO mice retarded the development of initial lung inflammation and subsequent emphysema, and this improvement correlated well with the appearance of macrophages expressing Nrf2-regulated antiprotease and antioxidant genes. Thus, Nrf2 appears to exert its protective effects through the transcriptional activation of antiprotease and antioxidant genes in alveolar macrophages. PMID- 16272358 TI - HIV-1-specific CD8+ T cell responses and viral evolution in women and infants. AB - CD8+ T lymphocyte responses play an important role in controlling HIV-1 replication but escape from CD8+ T cell surveillance may limit the effectiveness of these responses. Mother-to-child transmission of CD8+ T cell escape variants may particularly affect CD8+ T cell recognition of infant HIV-1 epitopes. In this study, amino acid sequence variation in HIV-1 gag and nef was examined in five untreated mother-infant pairs to evaluate the potential role of CD8+ T cell responses in the evolution of the viral quasispecies. Several CD8+ T cell escape variants were detected in maternal plasma. Evaluation of infant plasma viruses at 1-3 mo documented heterogeneity of gag and nef gene sequences and mother-to-child transmission of CD8+ T cell escape variants. Infant HLA haplotype and viral fitness appeared to determine the stability of the escape mutants in the infant over time. Changes in CD8+ T cell epitope sequences were detected in infants' sequential plasma specimens, suggesting that infants are capable of generating virus-specific CD8+ T cell responses that exert selective pressures in vivo. Altogether, these studies document that HIV-1-specific CD8+ T cell responses contribute to the evolution of the viral quasispecies in HIV-1-infected women and their infants and may have important implications for vaccine design. PMID- 16272360 TI - Stimulating lymphotoxin beta receptor on the dendritic cells is critical for their homeostasis and expansion. AB - The increased number of dendritic cells (DCs) inside lymphoid tissue may contribute to the enhanced priming of lymphocytes. The homeostasis of splenic DCs has mostly been attributed to their migration to the spleen via the chemokine microenvironment induced by lymphotoxin beta receptor (LTbetaR) signaling on splenic stromal cells. In this study we show that the lack of direct LTbetaR signaling on DCs is associated with the reduction of the number of DCs in the spleen independently of chemokine gradients. LTbetaR-/- mice have reduced DCs and reduced BrdU incorporation on DCs, and fewer DCs from LTbetaR-/- mice are detected in the spleen. Furthermore, increased expression of LIGHT (homologous to lymphotoxin, exhibits inducible expression, competes with herpesvirus glycoprotein D for herpes virus entry mediator on T cells) on T cells, a member of the TNF family (TNFSF14) and a ligand for LTbetaR, could dramatically increase the number of T cells and DCs, which leads to severe autoimmune diseases in a LTbetaR-dependent fashion. In vitro, LIGHT could directly promote accumulation of bone marrow-derived DCs. Furthermore, intratumor expression of LIGHT can dramatically expand DCs in situ, and inoculation of DCs into tumor tissues enhanced tumor immunity. Therefore, LTbetaR signaling on DCs is required for their homeostasis during physiology and pathological conditions, and increased LIGHT-LTbetaR interaction could stimulate DC expansion for T cell-mediated immunity. PMID- 16272359 TI - Inhibition of CX3CL1 (fractalkine) improves experimental autoimmune myositis in SJL/J mice. AB - Idiopathic inflammatory myopathy is a chronic inflammatory muscle disease characterized by mononuclear cell infiltration in the skeletal muscle. The infiltrated inflammatory cells express various cytokines and cytotoxic molecules. Chemokines are thought to contribute to the inflammatory cell migration into the muscle. We induced experimental autoimmune myositis (EAM) in SJL/J mice by immunization with rabbit myosin and CFA. In the affected muscles of EAM mice, CX3CL1 (fractalkine) was expressed on the infiltrated mononuclear cells and endothelial cells, and its corresponding receptor, CX3CR1, was expressed on the infiltrated CD4 and CD8 T cells and macrophages. Treatment of EAM mice with anti CX3CL1 mAb significantly reduced the histopathological myositis score, the number of necrotic muscle fibers, and infiltration of CD4 and CD8 T cells and macrophages. Furthermore, treatment with anti-CX3CL1 mAb down-regulated the mRNA expression of TNF-alpha, IFN-gamma, and perforin in the muscles. Our results suggest that CX3CL1-CX3CR1 interaction plays an important role in inflammatory cell migration into the muscle tissue of EAM mice. The results also point to the potential therapeutic usefulness of CX3CL1 inhibition and/or blockade of CX3CL1 CX3CR1 interaction in idiopathic inflammatory myopathy. PMID- 16272362 TI - Proteomic scan for tyrosinase peptide antigenic pattern in vitiligo and melanoma: role of sequence similarity and HLA-DR1 affinity. AB - Immune responses contribute to the pathogenesis of vitiligo and target melanoma sometimes associated with vitiligo-like depigmentation in some melanoma patients. We analyzed the sera from patients with vitiligo and cutaneous melanoma for reactivity toward tyrosinase peptide sequences 1) endowed with low level of similarity to human proteome, and 2) potentially able to bind HLA-DR1 Ags. We report that the tyrosinase autoantigen was immunorecognized with the same molecular pattern by sera from vitiligo and melanoma patients. Five autoantigen peptides composed the immunodominant anti-tyrosinase response: aa95 104FMGFNCGNCK; aa175-182 LFVWMHYY; aa176-190FVWMHYYVSMDALLG; aa222 236IQKLTGDENFTIPYW, and aa233-247 IPYWDWRDAEKCDIC. All of the five antigenic peptides were characterized by being (or containing) a sequence with low similarity level to the self proteome. Sera from healthy subjects were responsive to aa95-104FMGFNCGNCK, aa222-236IQKLTGDENFTIPYW, and aa233-247 IPYWDWRDAEKCDIC, but did not react with the aa175-182LFVWMHYY and aa176-190FVWMHYYVSMDALLG peptide sequences containing the copper-binding His180 and the oculocutaneous albinism I A variant position F176. Our results indicate a clear-cut link between peptide immunogenicity and low similarity level of the corresponding amino acid sequence, and are an example of a comparative analysis that might allow to comprehensively distinguish the epitopic peptide sequences within a disease from those associated to natural autoantibodies. In particular, these data, for the first time, delineate the linear B epitope pattern on tyrosinase autoantigen and provide definitive evidence of humoral immune responses against tyrosinase. PMID- 16272361 TI - Sodium stibogluconate interacts with IL-2 in anti-Renca tumor action via a T cell dependent mechanism in connection with induction of tumor-infiltrating macrophages. AB - IL-2 therapy results in 10-20% response rates in advanced renal cell carcinoma (RCC) via activating immune cells, in which the protein tyrosine phosphatase Src homology 2 domain-containing phosphatase 1 (SHP-1) is a key negative regulator. Based on finding that sodium stibogluconate (SSG) inhibited SHP-1, the anti-RCC potential and action mechanism of SSG and SSG/IL-2 in combination were investigated in a murine renal cancer model (Renca). Despite its failure to inhibit Renca cell proliferation in cultures, SSG induced 61% growth inhibition of Renca tumors in BALB/c mice coincident with an increase (2-fold) in tumor infiltrating macrophages (Mphi). A combination of SSG and IL-2 was more effective in inhibiting tumor growth (91%) and inducing tumor-infiltrating Mphi (4-fold), whereas IL-2 alone had little effect. Mphi increases were also detected in the spleens of mice treated with SSG (3-fold) or SSG/IL-2 in combination (6-fold), suggesting a systemic Mphi expansion similar to those in SHP-deficient mice. T cell involvement in the anti-Renca tumor action of the combination was suggested by the observations that the treatment induced spleen IFN-gamma T cells in BALB/c mice, but failed to inhibit Renca tumor growth in athymic nude mice and that SSG treatment of T cells in vitro increased production of IFN-gamma capable of activating tumoricidal Mphi. The SSG and SSG/IL-2 combination treatments were tolerated in the mice. These results together demonstrate an anti-Renca tumor activity of SSG that was enhanced in combination with IL-2 and functions via a T cell-dependent mechanism with increased IFN-gamma production and expansion/activation of Mphi. Our findings suggest that SSG might improve anti RCC efficacy of IL-2 therapy by enhancing antitumor immunity. PMID- 16272363 TI - Constitutive activation of p38 and ERK1/2 MAPKs in epithelial cells of myasthenic thymus leads to IL-6 and RANTES overexpression: effects on survival and migration of peripheral T and B cells. AB - Myasthenia gravis (MG) is an autoimmune disease of neuromuscular junctions where thymus plays a pathogenetic role. Thymectomy benefits patients, and thymic hyperplasia, a lymphoid infiltration of perivascular spaces becoming site of autoantibody production, is recurrently observed. Cytokines and chemokines, produced by thymic epithelium and supporting survival and migration of T and B cells, are likely to be of great relevance in pathogenesis of thymic hyperplasia. In thymic epithelial cell (TEC) cultures derived "in vitro" from normal or hyperplastic age-matched MG thymuses, we demonstrate by gene profiling analysis that MG-TEC basally overexpress genes coding for p38 and ERK1/2 MAPKs and for components of their signaling pathways. Immunoblotting experiments confirmed that p38 and ERK1/2 proteins were overexpressed in MG-TEC and, in addition, constitutively activated. Pharmacological blockage with specific inhibitors confirmed their role in the control of IL-6 and RANTES gene expression. According to our results, IL-6 and RANTES levels were abnormally augmented in MG-TEC, either basally or upon induction by adhesion-related stimuli. The finding that IL 6 and RANTES modulate, respectively, survival and migration of peripheral lymphocytes of myasthenic patients point to MAPK transcriptional and posttranscriptional abnormalities of MG-TEC as a key step in the pathological remodelling of myasthenic thymus. PMID- 16272364 TI - Beryllium presentation to CD4+ T cells is dependent on a single amino acid residue of the MHC class II beta-chain. AB - Chronic beryllium disease (CBD) is characterized by a CD4+ T cell alveolitis and granulomatous inflammation in the lung. Genetic susceptibility to this disease has been linked with HLA-DP alleles, particularly those possessing a glutamic acid at position 69 (Glu69) of the beta-chain. However, 15% of CBD patients do not possess a Glu69-containing HLA-DP allele, suggesting that other MHC class II alleles may be involved in disease susceptibility. In CBD patients without a Glu69-containing HLA-DP allele, an increased frequency of HLA-DR13 alleles has been described, and these alleles possess a glutamic acid at position 71 of the beta-chain (which corresponds to position 69 of HLA-DP). Thus, we hypothesized that beryllium presentation to CD4+ T cells was dependent on a glutamic acid residue at the identical position of both HLA-DP and -DR. The results show that HLA-DP Glu69- and HLA-DR Glu71-expressing molecules are capable of inducing beryllium-specific proliferation and IFN-gamma expression by lung CD4+ T cells. Using fibroblasts expressing mutated HLA-DP2 and -DR13 molecules, beryllium recognition was dependent on the glutamic acid at position 69 of HLA-DP and 71 of HLA-DR, suggesting a critical role for this amino acid in beryllium presentation to Ag-specific CD4+ T cells. Thus, these results demonstrate that a single amino acid residue of the MHC class II beta-chain dictates beryllium presentation and potentially, disease susceptibility. PMID- 16272365 TI - Gliadin fragments induce phenotypic and functional maturation of human dendritic cells. AB - Celiac disease is a chronic inflammatory disease developing in genetically predisposed individuals. Ingested gliadin, the triggering agent of the disease, can cross the epithelial barrier and elicit a harmful T cell-mediated immune response. Dendritic cells (DC) are supposed to play a pivotal role in shaping the immune response. The direction of the immune response toward immunity or tolerance depends on the stage of maturation and the functional properties of the DC. DC become fully functional APC upon maturation by various stimuli. We investigated the effect of a peptic digest of gliadin on the maturation of human monocyte-derived DC. Stimulation of cells with gliadin, in contrast with other tested food proteins, led to enhanced expression of maturation markers (CD80, CD83, CD86, and HLA-DR molecules) and increased secretion of chemokines and cytokines (mainly of IL-6, IL-8, IL-10, TNF-alpha, growth-related oncogene, MCP 1, MCP-2, macrophage-derived chemokine, and RANTES). Maturation was accompanied by a greater capacity to stimulate proliferation of allogeneic T cells and significantly reduced endocytic activity. Furthermore, gliadin-induced phosphorylation of members of three MAPK families (ERK1/2, JNK, and p38 MAPK) was demonstrated. The largest contribution of p38 MAPK was confirmed using its inhibitor SB203580, which markedly down-regulated the gliadin-triggered up regulation of maturation markers and cytokine production. Gliadin treatment also resulted in increased NF-kappaB/DNA binding activity of p50 and p65 subunits. Taken together, gliadin peptides can contribute to overcoming the stage of unresponsiveness of immature DC by inducing phenotypic and functional DC maturation, resulting in more efficient processing and presentation of gliadin peptides to specific T lymphocytes. PMID- 16272367 TI - Genome update: distribution of two-component transduction systems in 250 bacterial genomes. PMID- 16272368 TI - Clinical significance of seeding dispersal in biofilms. PMID- 16272366 TI - Telomere length of transferred lymphocytes correlates with in vivo persistence and tumor regression in melanoma patients receiving cell transfer therapy. AB - Recent studies have indicated that adoptive immunotherapy with autologous antitumor tumor-infiltrating lymphocytes (TILs) following nonmyeloablative chemotherapy mediates tumor regression in approximately 50% of treated patients with metastatic melanoma, and that tumor regression is correlated with the degree of persistence of adoptively transferred T cells in peripheral blood. These findings, which suggested that the proliferative potential of transferred T cells may play a role in clinical responses, led to the current studies in which telomere length as well as phenotypic markers expressed on the administered TILs were examined. TILs that were associated with objective clinical responses following adoptive transfer possessed a mean telomere length of 6.3 kb, whereas TILs that were not associated with significant clinical responses were significantly shorter, averaging 4.9 kb (p < 0.01). Furthermore, individual TIL derived T cell clonotypes that persisted in vivo following adoptive cell transfer possessed telomeres that were longer than telomeres of T cell clonotypes that failed to persist (6.2 vs 4.5 kb, respectively; p < 0.001). Expression of the costimulatory molecule CD28 also appeared to be associated with long telomeres and T cell persistence. These results, indicating that the telomere length of transferred lymphocytes correlated with in vivo T cell persistence following adoptive transfer, and coupled with the previous observation that T cell persistence was associated with clinical responses in this adoptive immunotherapy trial, suggest that telomere length and the proliferative potential of the transferred T cells may play a significant role in mediating response to adoptive immunotherapy. PMID- 16272370 TI - Peptidoglycan degradation by specialized lytic transglycosylases associated with type III and type IV secretion systems. AB - Specialized lytic transglycosylases are muramidases capable of locally degrading the peptidoglycan meshwork of Gram-negative bacteria. Specialized lytic transglycosylase genes are present in clusters encoding diverse macromolecular transport systems. This paper reports the analysis of selected members of the specialized lytic transglycosylase family from type III and type IV secretion systems. These proteins were analysed in vivo by assaying their ability to complement the DNA transfer defect of the conjugative F-like plasmid R1-16 lacking a functional P19 protein, the specialized lytic transglycosylase of this type IV secretion system. Heterologous complementation was accomplished using IpgF from the plasmid-encoded type III secretion system of Shigella sonnei and TrbN from the type IV secretion system of the conjugative plasmid RP4. In contrast, neither VirB1 proteins (Agrobacterium tumefaciens, Brucella suis) nor IagB (Salmonella enterica) could functionally replace P19. In vitro, IpgF, IagB, both VirB1 proteins, HP0523 (Helicobacter pylori) and P19 displayed peptidoglycanase activity in zymogram analyses. Using an established test system and a newly developed assay it was shown that IpgF degraded peptidoglycan in solution. IpgF was active only after removal of the chaperonin GroEL, which co purified with IpgF and inhibited its enzymic activity. A mutant IpgF protein in which the predicted catalytic amino acid, Glu42, was replaced by Gln, was completely inactive. IpgF-catalysed peptidoglycan degradation was optimal at pH 6 and was inhibited by the lytic transglycosylase inhibitors hexa-N acetylchitohexaose and bulgecin A. PMID- 16272371 TI - The putative lytic transglycosylase VirB1 from Brucella suis interacts with the type IV secretion system core components VirB8, VirB9 and VirB11. AB - VirB1-like proteins are believed to act as lytic transglycosylases, which facilitate the assembly of type IV secretion systems via localized lysis of the peptidoglycan. This paper presents the biochemical analysis of interactions of purified Brucella suis VirB1 with core components of the type IV secretion system. Genes encoding VirB1, VirB8, VirB9, VirB10 and VirB11 were cloned into expression vectors; the affinity-tagged proteins were purified from Escherichia coli, and analyses by gel filtration chromatography showed that they form monomers or homo-multimers. Analysis of protein-protein interactions by affinity precipitation revealed that VirB1 bound to VirB9 and VirB11. The results of bicistron expression experiments followed by gel filtration further supported the VirB1-VirB9 interaction. Peptide array mapping identified regions of VirB1 that interact with VirB8, VirB9 and VirB11 and underscored the importance of the C terminus, especially for the VirB1-VirB9 interaction. The binding sites were localized on a structure model of VirB1, suggesting that different portions of VirB1 may interact with other VirB proteins during assembly of the type IV secretion machinery. PMID- 16272372 TI - Molecular characterization of the Agrobacterium tumefaciens DNA transfer protein VirB6. AB - The VirB proteins of Agrobacterium tumefaciens assemble a T-pilus and a type IV secretion (T4S) apparatus for the transfer of DNA and proteins to plant cells. VirB6 is essential for DNA transfer and is a polytopic integral membrane protein with at least four membrane-spanning domains. VirB6 is postulated to function in T-pilus biogenesis and to be a component of the T4S apparatus. To identify amino acids required for VirB6 function, random mutations were introduced into virB6, and mutants that failed to complement a deletion in virB6 in tumour formation assays were isolated. Twenty-one non-functional mutants were identified, eleven of which had a point mutation that led to a substitution in a single amino acid. Characterization of the mutants indicated that the N-terminal large periplasmic domain and the transmembrane domain TM3 are required for VirB6 function. TM3 has an unusual sequence feature in that it is rich in bulky hydrophobic amino acids. This feature is found conserved in the VirB6 family of proteins. Studies on the effect of VirB6 on other VirB proteins showed that the octopine Ti-plasmid VirB6, unlike its nopaline Ti-plasmid counterpart, does not affect accumulation of VirB3 and VirB5, but has a strong negative effect on the accumulation of the VirB7 VirB7 dimer. Using indirect immunofluorescence microscopy the authors recently demonstrated that VirB6 localizes to a cell pole in a VirB-dependent manner. Mutations identified in the present study did not affect polar localization of the protein or the formation of the VirB7-VirB7 dimer. A VirB6-GFP fusion that contained the entire VirB6 ORF did not localize to a cell pole in either the presence or the absence of the other VirB proteins. IMF studies using dual labelling demonstrated that VirB6 colocalizes with VirB3 and VirB9, and not with VirB4, VirB5 and VirB11. These results support the conclusion that VirB6 is a structural component of the T4S apparatus. PMID- 16272373 TI - Conjugative plasmid DNA transfer in Helicobacter pylori mediated by chromosomally encoded relaxase and TraG-like proteins. AB - One of the striking characteristics of Helicobacter pylori is the extensive genetic diversity among clinical isolates. This diversity has been attributed to an elevated mutation rate, impaired DNA repair, DNA transfer and frequent recombination events. Plasmids have also been identified in H. pylori but it remained unknown whether conjugation can contribute to DNA transfer between clinical isolates. To examine whether H. pylori possesses intrinsic capability for conjugative plasmid transfer, shuttle vectors were introduced into H. pylori containing an oriT sequence of the conjugative IncPalpha plasmid RP4 but no mobilization (mob) genes. It was shown that these vectors could stably replicate and be mobilized among clinical H. pylori strains. It was also demonstrated that traG and relaxase (rlx) homologues carried on the H. pylori chromosome were important for plasmid transfer. Primer extension studies and mutagenesis further confirmed that the relaxase homologue rlx1 in H. pylori encodes a functional enzyme capable of acting on the RP4 oriT. Furthermore, the findings of this study indicate that traG and rlx1 act independently of the previously described type IV secretion systems, including that encoded by the cag pathogenicity island and the comB transformation apparatus, in mediating conjugative plasmid DNA transfer between H. pylori strains. PMID- 16272374 TI - Functional interactions between type IV secretion systems involved in DNA transfer and virulence. AB - This paper reports an analysis of the functional interactions between type IV secretion systems (T4SS) that are part of the conjugative machinery for horizontal DNA transfer (cT4SS), and T4SS involved in bacterial pathogenicity (pT4SS). The authors' previous work showed that a conjugative coupling protein (T4CP) interacts with the VirB10-type component of the T4SS in order to recruit the protein-DNA complex to the transporter for conjugative DNA transfer. This study now shows by two-hybrid analysis that conjugative T4CPs also interact with the VirB10 element of the pT4SS of Agrobacterium tumefaciens (At), Bartonella tribocorum (Bt) and Brucella suis (Bs). Moreover, the VirB10 component of a cT4SS (protein TrwE of plasmid R388) could be partially substituted by that of a pT4SS (protein TrwE of Bt) for conjugation. This result opens the way for the construction of hybrid T4SS that deliver DNA into animal cells. Interestingly, in the presence of part of the Bs T4SS the R388 T4SS protein levels were decreased and R388 conjugation was strongly inhibited. Complementation assays between the Trw systems of R388 and Bt showed that only individual components from the so called 'core complex' could be exchanged, supporting the concept that this core is the common scaffold for the transport apparatus while the other 'peripheral components' are largely system-specific. PMID- 16272375 TI - Unsaturated fatty acids are inhibitors of bacterial conjugation. AB - This report describes a high-throughput assay to identify substances that reduce the frequency of conjugation in Gram-negative bacteria. Bacterial conjugation is largely responsible for the spread of multiple antibiotic resistances in human pathogens. Conjugation inhibitors may provide a means to control the spread of antibiotic resistance. An automated conjugation assay was developed that used plasmid R388 and a laboratory strain of Escherichia coli as a model system, and bioluminescence as a reporter for conjugation activity. Frequencies of conjugation could be measured continuously in real time by the amount of light produced, and thus the effects of inhibitory compounds could be determined quantitatively. A control assay, run in parallel, allowed elimination of compounds affecting cell growth, plasmid stability or gene expression. The automated conjugation assay was used to screen a database of more than 12,000 microbial extracts known to contain a wide variety of bioactive compounds (the NatChem library). The initial hit rate was 1.4 %. From these, 48 extracts containing active compounds and representing a variety of organisms and extraction conditions were subjected to fractionation (24 fractions per extract). The 52 most active fractions were subjected to a secondary analysis to determine the range of plasmid inhibition. Plasmids R388, R1 and RP4 were used as representatives of a variety of plasmid transfer systems. Only one fraction (of complex composition) affected transfer of all three plasmids, while four other fractions were active against two of them. Two separate compounds were identified from these fractions: linoleic acid and dehydrocrepenynic acid. Downstream analysis showed that the chemical class of unsaturated fatty acids act as true inhibitors of conjugation. PMID- 16272376 TI - The mating pair stabilization protein, TraN, of the F plasmid is an outer membrane protein with two regions that are important for its function in conjugation. AB - F plasmid TraN (602 aa, processed to 584 aa with 22 conserved cysteines), which is essential for F plasmid conjugation, is an outer-membrane protein involved in mating pair stabilization (MPS). Unlike R100 TraN, F TraN requires OmpA in the recipient cell for efficient MPS. The authors have identified three external loops (aa 172-187, 212-220 and 281-284) in the highly divergent region from aa 164 to aa 333 as candidates for interaction with OmpA. These loops were identified using both site-directed and random TnphoA/in mutagenesis to insert epitopes (31-aa or c-myc) into TraN and monitor their effect on sensitivity to external proteases and on mating ability. TraN is a hallmark protein of F-type IV secretion systems as demonstrated by blast searches of the databases. The C terminal region is highly conserved and contains five of the six completely conserved cysteines. Mutation of these residues to serine demonstrated their importance in TraN function. TraN appears to require both intra- and intermolecular disulfide bond formation for its stability and structure as demonstrated by its instability in a dsbA mutant and its aberrant migration on SDS-polyacrylamide gels under non-reducing conditions or by cross-linking with bis(sulfosuccinimidyl)suberate (BS3). Thus, F TraN appears to have two domains: the N-terminal region is involved in OmpA interaction with OmpA during MPS; and the C-terminal region, which is rich in conserved cysteine residues, is essential for conjugation. PMID- 16272377 TI - Fluorescence assays for F-pili and their application. AB - Conjugative pili are extracellular filaments elaborated by Gram-negative bacteria expressing certain type IV secretion systems. They are required at the earliest stages of conjugal DNA transfer to establish specific and secure cell-cell contacts. Conjugative pili also serve as adsorption organelles for both RNA and DNA bacteriophages. Beyond these facts, the structure, formation and function of these filaments are poorly understood. This paper describes a rapid, quantitative assay for F-pili encoded by the F plasmid type IV secretion system. The assay is based on the specific lateral adsorption of icosahedral RNA bacteriophage R17 by F-pili. Bacteriophage particles conjugated with a fluorescent dye, Alexa 488, and bound to F-pili defined filaments visible by immunofluorescence microscopy. F pili attached to F+ cells and free F-pili were both visible by this method. For quantification, cell-bound bacteriophage were separated from free bacteriophage particles by sedimentation and released by suspending cell pellets in 0.1 % SDS. Fluorescence in cell-free supernatant fractions was measured by fluorometry. The authors present a characterization of this assay and its application to F-pilus formation by cells carrying mutations in the gene for the F-pilus subunit F pilin. Each mutation introduced a cysteine, which F-pilin normally lacks, at a different position in its primary structure. Cysteine residues in the N-terminal domain I abolished filament formation as measured by fluorescent R17 binding. This was confirmed by measurements of DNA donor activity and filamentous DNA bacteriophage infection. With one exception (G53C), cysteines elsewhere in the F pilin primary structure did not abolish filament formation, although some mutations differentially affected F-pilus functions. PMID- 16272378 TI - Subcellular localization and functional domains of the coupling protein, TraG, from IncHI1 plasmid R27. AB - Bacterial conjugation is a horizontal gene transfer event mediated by the type IV secretion system (T4SS) encoded by bacterial plasmids. Within the T4SS, the coupling protein plays an essential role in linking the membrane-associated pore forming proteins to the cytoplasmic, DNA-processing proteins. TraG is the coupling protein encoded by the incompatibility group HI plasmids. A hallmark feature of the IncHI plasmids is optimal conjugative transfer at 30 degrees C and an inability to transfer at 37 degrees C. Transcriptional analysis of the transfer region 1 (Tra1) of R27 has revealed that traG is transcribed in a temperature-dependent manner, with significantly reduced levels of expression at 37 degrees C as compared to expression at 30 degrees C. The R27 coupling protein contains nucleoside triphosphate (NTP)-binding domains, the Walker A and Walker B boxes, which are well conserved among this family of proteins. Site-specific mutagenesis within these motifs abrogated the conjugative transfer of R27 into recipient cells. Mutational analysis of the TraG periplasmic-spanning residues, in conjunction with bacterial two-hybrid and immunoprecipitation analysis, determined that this region is essential for a successful interaction with the T4SS protein TrhB. Further characterization of TraG by immunofluorescence studies revealed that the R27 coupling protein forms membrane-associated fluorescent foci independent of R27 conjugative proteins. These foci were found at discrete positions within the cell periphery. These results allow the definition of domains within TraG that are involved in conjugative transfer, and determination of the cellular location of the R27 coupling protein. PMID- 16272379 TI - Transcriptional organization of the temperature-sensitive transfer system from the IncHI1 plasmid R27. AB - One of the characteristic features of IncHI1 plasmids is a thermosensitive process of conjugation, which is optimal between 22 degrees C and 30 degrees C but inhibited at 37 degrees C. R27, the prototypical IncHI1 plasmid, contains transfer genes clustered in two regions of the plasmid, Tra1 and Tra2. In the present study, transcriptional analyses of the tra genes were undertaken at both 30 degrees C and 37 degrees C. Screening of 38 tra genes showed that tra genes are transcriptionally linked in six operons, three in each Tra region. RT-PCR analysis showed that gene expression was reduced at 37 degrees C relative to that observed at 30 degrees C. The transcription start sites of the six transcripts were identified, promoters and upstream regions were cloned, and transcription was tested at both temperatures. In cells grown at 37 degrees C, in the presence of R27, the promoters were inhibited, except for promoters of the H operon and AN operon. Conditions that influenced DNA topology, such as osmolarity, anaerobiosis, quorum sensing and acidity, showed no significant influence on transfer frequency. These results should facilitate future understanding of the basis of temperature-sensitive transfer in this large conjugative plasmid. PMID- 16272380 TI - Factors triggering type III secretion in Pseudomonas aeruginosa. AB - The type III secretion system of Pseudomonas aeruginosa is tightly regulated by various environmental signals, such as low calcium and contact with the host cell. However, the exact signals triggering type III secretion are unknown. The present study describes the finding that secretion of P. aeruginosa type III effector molecules requires protein factors from serum and L broth, designated type III secretion factors (TSFs), in addition to the low-calcium environment. In the absence of TSF or calcium chelator EGTA, basal levels of type III effector molecules are accumulated intracellularly. Addition of TSF and EGTA together effectively triggers the secretion of pre-existing effector molecules in a short time, even before the active expression of type III genes; thus, active type III gene expression does not seem to be a prerequisite for type III secretion. A search for TSF molecules in serum and L broth resulted in the identification of albumin and casein as the functional TSF molecules. Although there is no clear sequence similarity between albumin and casein, both proteins are known to have a low-affinity, high-capacity calcium-binding property. Tests of well-studied calcium-binding proteins seemed to indicate that low-affinity calcium-binding proteins have TSF activity, although the requirement of low-affinity calcium binding ability for the TSF activity is not clear. P. aeruginosa seems to have evolved a sensing mechanism to detect target cells for type III injection through host-derived proteins in combination with a low-calcium signal. Disruption of the bacterial ability to sense low calcium or TSF might be a valid avenue to the effective control of this bacterial pathogen. PMID- 16272381 TI - The LuxR receptor: the sites of interaction with quorum-sensing signals and inhibitors. AB - The function of LuxR homologues as quorum sensors is mediated by the binding of N acyl-L-homoserine lactone (AHL) signal molecules to the N-terminal receptor site of the proteins. In this study, site-directed mutagenesis was carried out of the amino acid residues comprising the receptor site of LuxR from Vibrio fischeri, and the ability of the L42A, L42S, Y62F, W66F, D79N, W94D, V109D, V109T and M135A LuxR mutant proteins to activate green fluorescent protein expression from a P(luxI) promoter was measured. X-ray crystallographic studies of the LuxR homologue TraR indicated that residues Y53 and W57 form hydrogen bonds to the 1 carbonyl group and the ring carbonyl group, respectively, of the cognate AHL signal. Based on the activity and signal specificity of the LuxR mutant proteins, and on molecular modelling, a model is suggested in which Y62 (corresponding to Y53 in TraR) forms a hydrogen bond with the ring carbonyl group rather than the 1 carbonyl group, while W66 (corresponding to W57 in TraR) forms a hydrogen bond to the 1-carbonyl group. This flips the position of the acyl side chain in the LuxR/signal molecule complex compared to the TraR/signal molecule complex. Halogenated furanones from the marine alga Delisea pulchra and the synthetic signal analogue N-(sulfanylacetyl)-L-homoserine lactone can block quorum sensing. The LuxR mutant proteins were insensitive to inhibition by N (propylsulfanylacetyl)-L-homoserine lactone. In contrast, the mutations had only a minor effect on the sensitivity of the proteins to halogenated furanones, and the data strongly suggest that these compounds do not compete in a 'classic' way with N-3-oxohexanoyl-L-homoserine lactone for the binding site. Based on modelling and experimental data it is suggested that these compounds bind in a non-agonist fashion. PMID- 16272382 TI - Identification of a novel two-component system SenS/SenR modulating the production of the catalase-peroxidase CpeB and the haem-binding protein HbpS in Streptomyces reticuli. AB - The Gram-positive soil bacterium and cellulose degrader Streptomyces reticuli synthesizes the mycelium-associated enzyme CpeB, which displays haem-dependent catalase and peroxidase activity, as well as haem-independent manganese peroxidase activity. The expression of the furS-cpeB operon depends on the redox regulator FurS and the presence of the haem-binding protein HbpS. Upstream of hbpS, the neighbouring senS and senR genes were identified. SenS is a sensor histidine kinase with five predicted N-terminally located transmembrane domains. SenR is the corresponding response regulator with a C-terminal DNA-binding motif. Comparative transcriptional and biochemical studies with a designed S. reticuli senS/senR chromosomal disruption mutant and a set of constructed Streptomyces lividans transformants showed that the presence of the novel two-component system SenS/SenR negatively modulates the expression of the furS-cpeB operon and the hbpS gene. The presence of SenS/SenR enhances considerably the resistance of S. reticuli to haemin and the redox-cycling compound plumbagin, suggesting that this system could participate directly or indirectly in the sensing of redox changes. Epitope-tagged HbpS (obtained from an Escherichia coli transformant) as well as the native S. reticuli HbpS interact in vitro specifically with the purified SenS fusion protein. On the basis of these findings, together with data deduced from the S. reticuli hbpS mutant strain, HbpS is suggested to act as an accessory protein that communicates with the sensor protein to modulate the corresponding regulatory cascade. Interestingly, close and distant homologues, respectively, of the SenS/SenR system are encoded within the Streptomyces coelicolor A3(2) and Streptomyces avermitilis genomes, but not within other known bacterial genomes. Hence the SenS/SenR system appears to be confined to streptomycetes. PMID- 16272383 TI - The role of two CbbRs in the transcriptional regulation of three ribulose-1,5 bisphosphate carboxylase/oxygenase genes in Hydrogenovibrio marinus strain MH 110. AB - Hydrogenovibrio marinus MH-110 possesses three different sets of genes for ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO): two form I (cbbLS-1 and cbbLS-2) and one form II (cbbM). We have previously shown that the expression of these RubisCO genes is dependent on the ambient CO2 concentration. LysR-type transcriptional regulators, designated CbbR1 and CbbRm, are encoded upstream of the cbbLS-1 and cbbM genes, respectively. In this study, we revealed by gel shift assay that CbbR1 and CbbRm bind with higher affinity to the promoter regions of cbbLS-1 and cbbM, respectively, and with lower affinity to the other RubisCO gene promoters. The expression patterns of the three RubisCOs in the cbbR1 and the cbbRm gene mutants showed that CbbR1 and CbbRm were required to activate the expression of cbbLS-1 and cbbM, respectively, and that neither CbbR1 nor CbbRm was required for the expression of cbbLS-2. The expression of cbbLS-1 was significantly enhanced under high-CO2 conditions in the cbbRm mutant, in which the expression of cbbM was decreased. Although cbbLS-2 was not expressed under high-CO2 conditions in the wild-type strain or the single cbbR mutants, the expression of cbbLS-2 was observed in the cbbR1 cbbRm double mutant, in which the expression of both cbbLS-1 and cbbM was decreased. These results indicate that there is an interactive regulation among the three RubisCO genes. PMID- 16272384 TI - ESCRT-I components of the endocytic machinery are required for Rim101-dependent ambient pH regulation in the yeast Yarrowia lipolytica. AB - Ambient pH signalling involves a cascade of conserved Rim or Pal products in ascomycetous yeasts or filamentous fungi, respectively. Insertional mutagenesis in the yeast Yarrowia lipolytica identified two components of the endosome associated ESCRT-I complex involved in multivesicular body (MVB) vesicle formation, YlVps28p and YlVps23p. They were shown to be required at alkaline pH, like Rim factors, for transcriptional activation of alkaline-induced genes and repression of acid-induced genes. The constitutively active YlRIM101-1119 allele, which suppresses the pH-signalling defects of Ylrim mutations, also suppresses Ylvps defects in pH response, but not in endocytosis. The contribution of the ESCRT-III component Snf7p could not be assessed due to the essential nature of this component in Y. lipolytica. Unlike Rim factors, YlVps4p, a component of the MVB pathway acting downstream from ESCRT complexes, seems not to be required for the alkaline response. In Y. lipolytica, all vps mutations including those affecting YlVPS4, affected growth at acidic pH, a feature not exhibited by Ylrim mutations. These results suggest that Rim and Vps pathways cooperate in ambient pH signalling and that this relation is conserved across the full range of hemiascomycetous yeasts. PMID- 16272385 TI - Nitrilase from Pseudomonas fluorescens EBC191: cloning and heterologous expression of the gene and biochemical characterization of the recombinant enzyme. AB - The gene encoding an enantioselective arylacetonitrilase was identified on a 3.8 kb DNA fragment from the genomic DNA of Pseudomonas fluorescens EBC191. The gene was isolated, sequenced and cloned into the L-rhamnose-inducible expression vector pJOE2775. The nitrilase was produced in large quantities and purified as a histidine-tagged enzyme from crude extracts of L-rhamnose-induced cells of Escherichia coli JM109. The purified nitrilase was significantly stabilized during storage by the addition of 1 M ammonium sulfate. The temperature optimum (50 degrees C), pH optimum (pH 6.5), and specific activity of the recombinant nitrilase were similar to those of the native enzyme from P. fluorescens EBC191. The enzyme hydrolysed various phenylacetonitriles with different substituents in the 2-position and also heterocyclic and bicyclic arylacetonitriles to the corresponding carboxylic acids. The conversion of most arylacetonitriles was accompanied by the formation of different amounts of amides as by-products. The relative amounts of amides formed from different nitriles increased with an increasing negative inductive effect of the substituent in the 2-position. The acids and amides that were formed from chiral nitriles demonstrated in most cases opposite enantiomeric excesses. Thus mandelonitrile was converted by the nitrilase preferentially to R-mandelic acid and S-mandelic acid amide. The nitrilase gene is physically linked in the genome of P. fluorescens with genes encoding the degradative pathway for mandelic acid. This might suggest a natural function of the nitrilase in the degradation of mandelonitrile or similar naturally occurring hydroxynitriles. PMID- 16272386 TI - Methylcrotonyl-CoA and geranyl-CoA carboxylases are involved in leucine/isovalerate utilization (Liu) and acyclic terpene utilization (Atu), and are encoded by liuB/liuD and atuC/atuF, in Pseudomonas aeruginosa. AB - Pseudomonas aeruginosa is able to grow on acyclic monoterpenes (citronellol, citronellate, geraniol and geranylate), and on other methyl-branched compounds such as leucine or isovalerate. The catabolic pathway of citronellol (Atu, acyclic terpene utilization) enters that of leucine/isovalerate (Liu, leucine and isovalerate utilization) at the level of methylcrotonyl-CoA. Key enzymes of the combined pathways are geranyl-CoA carboxylase (GCase) and methylcrotonyl-CoA carboxylase (MCase). In this study, isovalerate-grown cells specifically expressed MCase (apparent molecular mass of the biotin-containing subunit, 74 kDa) only, and the GCase biotin-containing subunit (71 kDa) was not detected. Citronellol- or citronellate-grown cells produced both carboxylases. Biotin dependent proteins were purified from crude extracts by avidin-affinity chromatography, and assigned to the corresponding coding genes by trypsin fingerprint analysis. The two subunits of MCase corresponded to liuB/liuD (PA2014/PA2012) of the P. aeruginosa genome database, and atuC/atuF (PA2888/PA2891) encoded GCase subunits. This finding is contrary to that reported by others. The identified genes are part of two separate gene clusters [liuRABCDE (PA2011-PA2016) and atuABCDEFGH (PA2886-PA2893)] that are thought to encode most of the genes of the Atu and Liu pathways. PMID- 16272387 TI - Protein O-mannosyltransferase A of Aspergillus awamori is involved in O mannosylation of glucoamylase I. AB - Industrially important extracellular enzymes from filamentous fungi are often O mannosylated. The structure and function of the pmtA (AapmtA) gene encoding the protein O-D-mannosyltransferase of Aspergillus awamori were characterized. The AapmtA disruptant, designated AaPMTA, was constructed by homologous recombination. The strain AaPMTA exhibited fragile cell morphology with respect to hyphal extension, as well as swollen hyphae formation and conidia formation in potato dextrose medium. Moreover, the AapmtA disruptant showed increased sensitivity to high temperature and Congo red. Thus, the AaPmtA protein is involved in the formation of the normal cell wall. The strain AaPMTA could grow well in liquid synthetic medium and secrete glucoamylase I (GAI-AaPMTA) to a similar extent to the wild-type strain (GAI-WT). Matrix-assisted laser desorption ionization time-of-flight mass spectrometry analysis of the GAIs revealed that approximately 33 mannose moieties of GAI were absent in strain AaPMTA. This result indicates that the AaPmtA protein is responsible for the transfer of mannose to GAI. Structural analysis of the O-linked oligosaccharides of GAI also demonstrated that the AapmtA disruption resulted in a reduction of the amounts of O-linked oligosaccharides, such as D-mannose and alpha-1,2-mannotriose, in GAI AaPMTA. However, the amount of alpha-1,2-mannobiose was comparable between GAI-WT and GAI-AaPMTA. The result suggests the presence of a compensatory mechanism in the synthetic pathway of O-mannosylation in A. awamori. PMID- 16272388 TI - Knockdown of LIM15/DMC1 in the mushroom Coprinus cinereus by double-stranded RNA mediated gene silencing. AB - The basidiomycete Coprinus cinereus has many advantages as a model organism for studying sexual development and meiosis, but it has been difficult to investigate using reverse-genetics methods, such as gene disruption by homologous recombination. Here, gene repression by dsRNA-mediated gene silencing was tried as an alternative method for reverse-genetics studies. It was shown that transformation of the LIM15/DMC1 dsRNA expression construct (LIM15dsRNA) resulted in genomic insertion of LIM15dsRNA and paucity of the LIM15/DMC1 transcript. First, LIM15dsRNA was transformed into the homothallic strain AmutBmut to generate a homozygote in which both nuclei had a copy of LIM15dsRNA. The LIM15/DMC1-repressed strain showed abnormal homologous chromosome synapsis during meiosis. Basidiospore production was reduced to 16 % by the induction of dsRNA. However, approximately 60 % of basidiospores were viable. Next, a heterozygote was generated in which one nucleus had a copy of LIM15dsRNA. The phenotype was similar to that of the homozygote. These results are not only the first demonstration of dsRNA-mediated gene silencing in a member of the homobasidiomycete fungi, to which 90 % of mushroom species belong, but also the first successful use of a reverse-genetics approach in C. cinereus research. PMID- 16272389 TI - Surface ultrastructure and elasticity in growing tips and mature regions of Aspergillus hyphae describe wall maturation. AB - This study reports the first direct, high-resolution physical and structural evidence of wall changes during hyphal tip growth, visualized by atomic force microscopy (AFM) in Aspergillus nidulans. Images from AFM and cryo-scanning electron microscopy provided comparable information, but AFM was also able to image and physically probe living cells. AFM images showed changes in the surface ultrastructure of A. nidulans hyphae, from newly deposited walls at hyphal tips to fully mature walls, as well as additional changes at young branches arising from mature walls. Surface architecture during wall maturation correlated with changes in the relative viscoelasticity (compliance per unit applied force) of walls measured by force spectroscopy (FS) in growing A. nidulans hyphae. Growing tips showed greater viscoelasticity than mature walls, despite equal support from turgor. Branch tips had comparable viscoelasticity to hyphal tips, unlike the mature wall from which they grew. FS also revealed differences in surface hydrophilicity between newly deposited and mature walls, with the tips being more hydrophilic. The hydrophilicity of young branch tips was similar to that of hyphal tips, and different from that of mature walls. Taken together, AFM images and FS data suggest that the A. nidulans wall matures following deposition at the hyphal tip. PMID- 16272390 TI - A death round affecting a young compartmentalized mycelium precedes aerial mycelium dismantling in confluent surface cultures of Streptomyces antibioticus. AB - Development-associated cell-death processes were investigated in detail during the growth and differentiation of Streptomyces antibioticus ATCC 11891 on confluent surface cultures, by using fluorescent viability probes, membrane and activity fluorescence indicators, and electron microscopy analysis. A previously unsuspected complexity was revealed, namely the presence of a very young compartmentalized mycelium that dies following an orderly pattern, leaving alternating live and dead segments in the same hypha. This death round is followed by the growth of a second mycelium which develops rapidly from the live segments of the first mycelium and dies massively in a second death round, which extends over the phases of aerial mycelium formation and sporulation. PMID- 16272392 TI - The fluorene catabolic linear plasmid in Terrabacter sp. strain DBF63 carries the beta-ketoadipate pathway genes, pcaRHGBDCFIJ, also found in proteobacteria. AB - Terrabacter sp. strain DBF63 is capable of degrading fluorene (FN) to tricarboxylic acid cycle intermediates via phthalate and protocatechuate. Genes were identified for the protocatechuate branch of the beta-ketoadipate pathway (pcaR, pcaHGBDCFIJ) by sequence analysis of a 70 kb DNA region of the FN catabolic linear plasmid pDBF1. RT-PCR analysis of RNA from DBF63 cells grown with FN, dibenzofuran, and protocatechuate indicated that the pcaHGBDCFIJ operon was expressed during both FN and protocatechuate degradation in strain DBF63. The gene encoding beta-ketoadipate enol-lactone hydrolase (pcaD) was not fused to the next gene, which encodes gamma-carboxymuconolactone decarboxylase (pcaC), in strain DBF63, even though the presence of the pcaL gene (the fusion of pcaD and pcaC) within a pca gene cluster has been thought to be a Gram-positive trait. Quantitative RT-PCR analysis revealed that pcaD mRNA levels increased sharply in response to protocatechuate, and a biotransformation experiment with cis,cis muconate using Escherichia coli carrying both catBC and pcaD indicated that PcaD exhibited beta-ketoadipate enol-lactone hydrolase activity. The location of the pca gene cluster on the linear plasmid, and the insertion sequences around the pca gene cluster suggest that the ecologically important beta-ketoadipate pathway genes, usually located chromosomally, may be spread widely among bacterial species via horizontal transfer or transposition events. PMID- 16272391 TI - The trans-acting flagellar regulatory proteins, FliX and FlbD, play a central role in linking flagellar biogenesis and cytokinesis in Caulobacter crescentus. AB - The FliX/FlbD-dependent temporal transcription of late flagellar genes in Caulobacter crescentus requires the assembly of an early, class II-encoded flagellar structure. Class II flagellar-mutant strains exhibit a delay in the completion of cell division, with the accumulation of filamentous cells in culture. It is shown here that this cell-division defect is attributable to an arrest in the final stages of cell separation. Normal cell morphology could be restored in class II mutants by gain-of-function alleles of FliX or FlbD, suggesting that the timely completion of cell division requires these trans acting factors. In synchronized cultures, inhibition of cell division by depleting FtsZ resulted in normal initial expression of the late, FlbD-dependent fliK gene; however, the cell cycle-regulated cessation of transcription was delayed, indicating that cell division may be required to negatively regulate FlbD activity. Interestingly, prolonged depletion of FtsZ resulted in an eventual loss of FlbD activity that could be bypassed by a constitutive mutant of FlbD, but not of FliX, suggesting the possible existence of a second cell cycle dependent pathway for FlbD activation. PMID- 16272393 TI - Identification of an upstream regulatory sequence that mediates the transcription of mox genes in Methylobacterium extorquens AM1. AB - A multiple A-tract sequence has been identified in the promoter regions for the mxaF, pqqA, mxaW, mxbD and mxcQ genes involved in methanol oxidation in Methylobacterium extorquens AM1, a facultative methylotroph. Site-directed mutagenesis was exploited to delete or change this conserved sequence. Promoter xylE transcriptional fusions were used to assess promoter activity in these mutants. A fiftyfold drop in the XylE activity was observed for the mxaF and pqqA promoters without this sequence, and a five- to sixfold drop in the XylE activity was observed for the mxbD and mxcQ promoters without this sequence. Mutants were generated in the chromosomal copies in which this sequence was either deleted or altered, and these mutants were unable to grow on methanol. When one of these sequences was added to Plac of Escherichia coli, which is a weak constitutive promoter in M. extorquens AM1, the activity increased two- to threefold. These results suggest that this sequence is essential for normal expression of these genes in M. extorquens AM1, and may serve as a general enhancer element for genetic constructs in this bacterium. PMID- 16272394 TI - Genetic interaction of the SMC complex with topoisomerase IV in Bacillus subtilis. AB - The role of topoisomerase IV (Topo IV) and of the structural maintenance of chromosomes (SMC) complex in chromosome compaction and in global protein synthesis was investigated. Lowering of the levels of Topo IV led to chromosome decondensation, while overproduction induced chromosome hyper-compaction, showing that Topo IV has an influence on the compaction of the whole chromosome, in a manner similar to that of the SMC protein, though different in mechanism. Increased synthesis of Topo IV in smc-deleted cells partially rescued the growth and condensation defect of the deletion, but not the segregation defect, revealing that the two systems interact at a genetic level. Two-dimensional gel investigations showed that global protein synthesis is highly aberrant in smc deleted cells, and, to a different extent, also in cells lacking ScpA or ScpB, which form the SMC complex together with SMC protein. Overproduction of Topo IV partially rescued the defect in protein synthesis in smc mutant cells, indicating that Topo IV can restore the loss of negative supercoiling caused by the absence of SMC protein, but does not fully rescue the segregation defect. The data also show that the SMC protein has a dual function, in chromosome supercoiling and in active segregation. PMID- 16272395 TI - Production of anti-neurotoxin antibody is enhanced by two subcomponents, HA1 and HA3b, of Clostridium botulinum type B 16S toxin-haemagglutinin. AB - Clostridium botulinum type B strain produces two forms of progenitor toxin, 16S and 12S. The 12S toxin is formed by association of a neurotoxin (NTX) and a non toxic non-haemagglutinin (NTNH), and the 16S toxin is formed by conjugation of the 12S toxin with a haemagglutinin (HA). HA consists of four subcomponents designated HA1, HA2, HA3a and HA3b. When mice were immunized with formalin detoxified NTX, 12S or 16S, a significantly greater amount of anti-NTX antibody (Ab) was produced in the mice injected with 16S than in NTX- or 12S-injected mice. Immunization with NTX mixed with HA1 and/or HA3b also increased the anti NTX Ab production, whereas NTX mixed with HA2 did not, indicating that HA1 and HA3b have adjuvant activity. This was further confirmed by immunizing mice with human albumin (Alb) alone or Alb mixed with either HA1 or HA3b. When mouse-spleen cells were stimulated with NTX, 16S or different HA subcomponents, 16S, HA1, HA3b and the mixture of HA1 and HA3 significantly increased interleukin 6 (IL6) production compared with NTX alone. Transcription of IL6 mRNA was low after stimulation with NTX alone, but increased to 16S-stimulation levels when NTX was mixed with HA1 or HA3b. In flow cytometry using labelled Abs against CD3 and CD19, the percentage of CD19 cells was higher following stimulation with 16S or NTX mixed with HA1 or HA3b compared with stimulation with NTX. The percentage of CD3 cells remained unchanged. These results suggest strongly that HA1 and HA3b demonstrate adjuvant activity via increasing IL6 production. PMID- 16272396 TI - The streptococcal iron uptake (Siu) transporter is required for iron uptake and virulence in a zebrafish infection model. AB - A limited understanding of iron uptake mechanisms is available for Streptococcus pyogenes, a haemolytic human pathogen capable of using a variety of haemoproteins in addition to ferric and ferrous iron. This study characterizes a transporter named siu (for streptococcal iron uptake), which consists of an ATP-binding protein (SiuA), a substrate-binding protein (SiuD), and two membrane permease subunits (SiuBG). An siuG mutant was constructed and characterized. The mutant demonstrated growth reduction in comparison to the parent strain when grown in complex medium containing iron in the form of blood, haemoglobin or serum. Only a small reduction in the growth of the siuG mutant was observed in medium containing ferric iron. However, in iron uptake assays the siuG mutant showed a decrease of approximately 30 % in Fe3+ incorporation. Addition of 6 microM haem to the medium inhibited Fe3+ uptake by the wild-type by 76 %, while addition of protoporphyrin IX did not, suggesting that utilization of haem as an iron source is responsible for the inhibition of Fe3+ uptake. Inactivation of siuG moderately reduced the ability of haem to inhibit Fe3+ incorporation by the cells. Inactivation of siaB (encoding a membrane permease of a second iron transporter) had a similar outcome, and inactivation of both transporters had a cumulative effect. These observations implicate both the siu and sia transporters in haem utilization by Strep. pyogenes. Studies in a zebrafish infection model revealed that the siuG mutant was attenuated in both intramuscular and intraperitoneal routes of infection. Together these observations show that the siu system is an iron acquisition pathway in Strep. pyogenes that is important both in vitro and in vivo. PMID- 16272397 TI - Evaluation of O-antigen inactivation on Pla activity and virulence of Yersinia pseudotuberculosis harbouring the pPla plasmid. AB - Yersinia pestis is a species that emerged recently from Yersinia pseudotuberculosis and gained an exceptional pathogenicity potential. Among the major genetic differences between the plague bacillus and its ancestor is the acquisition of the pPla plasmid, which has been associated with the increased virulence of Y. pestis. In a previous study, introduction of pPla into Y. pseudotuberculosis did not lead to any modification of the virulence of the host bacterium. However, it was subsequently demonstrated that the presence of smooth lipopolysaccharide (LPS) inhibits the activity of Pla. In this study, pPla was introduced into a Y. pseudotuberculosis strain expressing smooth LPS, and into a variant in which a mutation that abrogates the formation of O-antigen (O-Ag) repeats (as in natural isolates of Y. pestis) was generated. It was found that in both strains, Pla was synthesized, exported to the bacterial membrane and processed as in Y. pestis. However, the ability of Pla to activate plasminogen was weak and observed only at 37 degrees C in the smooth strain, while this activity was similar to that of Y. pestis and expressed at both 28 and 37 degrees C in the O-Ag mutant strain. Similarly, Pla-mediated inactivation of the antiprotease alpha2-antiplasmin was not detected in the smooth Y. pseudotuberculosis strain grown at 28 degrees C, but was expressed at both temperatures in the O-Ag mutant strain. Despite the more efficient activity of Pla, the Y. pseudotuberculosis O-Ag mutant strain exhibited a lower pathogenicity upon subcutaneous infection of mice. The results thus indicate that, although abrogation of O side chain synthesis in a Y. pseudotuberculosis strain harbouring pPla potentiates the two proteolytic activities of Pla, this is not sufficient to confer to Y. pseudotuberculosis a higher pathogenicity potential. These results also suggest that acquisition of pPla may not have been sufficient to confer an immediate higher pathogenic potential to the ancestor Y. pestis strain. PMID- 16272398 TI - Regulation of expression of type I signal peptidases in Listeria monocytogenes. AB - The role of type I signal peptidases (SPases I) is to remove the signal peptides of preproteins exported by the general secretory pathway. The genome of Listeria monocytogenes contains a locus encoding three contiguous SPases I (denoted SipX, SipY and SipZ). The authors recently showed that SipX and SipZ perform distinct functions in protein secretion and bacterial pathogenicity. Here, the regulation of sip gene expression in broth and in infected eukaryotic cells was studied. The results show that expression of the three sip genes is (i) controlled by two distinct promoter regions that respond differently to growth phase and temperature variations, and (ii) influenced by PrfA (the transcriptional activator regulating most of the virulence genes of L. monocytogenes) and the stress proteins ClpC and ClpP. It was found that sip gene expression was strongly upregulated upon infection of eukaryotic cells when bacteria were still entrapped in the phagosomal compartment. This upregulation is compatible with the need of L. monocytogenes to optimize its production of virulence factors in the early stage of the intracellular cycle. PMID- 16272399 TI - YvcK of Bacillus subtilis is required for a normal cell shape and for growth on Krebs cycle intermediates and substrates of the pentose phosphate pathway. AB - The HPr-like protein Crh has so far been detected only in the bacillus group of bacteria. In Bacillus subtilis, its gene is part of an operon composed of six ORFs, three of which exhibit strong similarity to genes of unknown function present in many bacteria. The promoter of the operon was determined and found to be constitutively active. A deletion analysis revealed that gene yvcK, encoded by this operon, is essential for growth on Krebs cycle intermediates and on carbon sources metabolized via the pentose phosphate pathway. In addition, cells lacking YvcK acquired media-dependent filamentous or L-shape-like aberrant morphologies. The presence of high magnesium concentrations restored normal growth and cell morphology. Furthermore, suppressor mutants cured from these growth defects appeared spontaneously with a high frequency. Such suppressing mutations were identified in a transposon mutagenesis screen and found to reside in seven different loci. Two of them mapped in genes of central carbon metabolism, including zwf, which encodes glucose-6-phosphate dehydrogenase and cggR, the product of which regulates the synthesis of glyceraldehyde-3-phosphate dehydrogenase. All these results suggest that YvcK has an important role in carbon metabolism, probably in gluconeogenesis required for the synthesis of cell wall precursor molecules. Interestingly, the Escherichia coli homologous protein, YbhK, can substitute for YvcK in B. subtilis, suggesting that the two proteins have been functionally conserved in these different bacteria. PMID- 16272400 TI - Acetate excretion during growth of Salmonella enterica on ethanolamine requires phosphotransacetylase (EutD) activity, and acetate recapture requires acetyl-CoA synthetase (Acs) and phosphotransacetylase (Pta) activities. AB - This report shows that Salmonella enterica catabolizes ethanolamine to acetyl-CoA (Ac-CoA), which enters the glyoxylate bypass and tricarboxylic acid cycle for the generation of energy and central metabolites. During growth on ethanolamine, S. enterica excreted acetate, whose recapture depended on Ac-CoA synthetase (Acs) and the housekeeping phosphotransacetylase (Pta) enzyme activities. The Pta enzyme did not play a role in acetate excretion during growth of S. enterica on ethanolamine. It is proposed that during growth on ethanolamine, acetate excretion is necessary to maintain a pool of free CoA. Acetate excretion requires the eut operon-encoded phosphotransacetylase (EutD) and acetate kinase (Ack) enzymes. EutD function was not required for growth on ethanolamine, and an eutD strain showed only a slight reduction in growth rate. The existence of an as-yet unidentified system that releases acetate was revealed during growth of a strain lacking Acs, the housekeeping phosphotransacetylase (Pta), and EutD. The functions of pyruvate oxidase (PoxB), Ack and STM3118 protein [a homologue of the Saccharomyces cerevisiae Ac-CoA hydrolase (Ach1p) enzyme] were not involved in the release of acetate by the acs pta eutD strain. PMID- 16272401 TI - Regulation of nitrate reductase activity in Mycobacterium tuberculosis by oxygen and nitric oxide. AB - Nitrate reduction by Mycobacterium tuberculosis is regulated by control of the transport of nitrate into the cell by NarK2. When oxygen was introduced into hypoxic cultures, nitrite production was quickly inhibited. The nitrate-reducing enzyme itself is relatively insensitive to oxygen, suggesting that the inhibition of nitrite production by oxygen was a result of interference with nitrate transport. This was not due to degradation of NarK2, as the inhibition was reversed by the removal of oxygen although chloramphenicol prevented new synthesis of NarK2. The oxidant potassium ferricyanide was added to anaerobic cultures to produce a positive redox potential in the absence of oxygen. Nitrite production decreased, signifying that oxidizing conditions, rather than oxygen itself, were responsible for the inhibition of nitrate transport. Nitric oxide added to cultures allowed NarK2 to be active even in the presence of oxygen. A similar result was obtained with hydroxylamine and ethanol, both of which interfere with oxygen utilization and the electron transport chain. It is proposed that NarK2 senses the redox state of the cell, possibly by monitoring the flow of electrons to cytochrome oxidase, and adjusts its activity so that nitrate is transported under reducing, but not under oxidizing, conditions. PMID- 16272402 TI - Evaluation of inhibitory potencies for compounds inhibiting P-glycoprotein but without maximum effects: f2 values. AB - In cell culture systems with aqueous buffers, concentration-response curves to lipophilic inhibitors are difficult to establish because plateau effects (Imax) are often not reached because of limited drug solubility. Consequently, the inhibitory potency of a compound will not be definable using IC50 values (concentration exerting 50% of Imax). Since alternative potency measures f2 values, the concentrations required to double baseline signals have been proposed. Using both methods, we reevaluated the concentration-response curves of calcein assays with 78 compounds in three different cell culture systems and found a close correlation between both methods (r(s) = 0.93-0.99, p < or = 0.0028). These findings suggest that f2 values are a valuable alternative to define rank orders of highly lipophilic inhibitors as a basis for the prediction of pharmacological interaction properties in clinical settings. Although it was only tested for inhibition of P-glycoprotein, it seems likely that this method may be transferred to other assays with other proteins. PMID- 16272403 TI - Interspecies differences in pharmacokinetics and metabolism of S-3-(4-acetylamino phenoxy)-2-hydroxy-2-methyl-N-(4-nitro-3-trifluoromethylphenyl)-propionamide: the role of N-acetyltransferase. AB - N-Acetyltransferase (NAT) is one of the major phase II enzymes involved in drug metabolism. Both species differences and polymorphism are observed in NAT expression. During the preclinical development of a novel selective androgen receptor modulator, S-3-(4-acetylamino-phenoxy)-2-hydroxy-2-methyl-N-(4-nitro-3 trifluoromethyl-phenyl)-propionamide (S4), we also observed species differences in S4 metabolism due to the interaction between the deacetylation metabolite M1 and NAT, which converted M1 back to S4 both in vitro and in vivo. During incubation with human liver cytosol or rat liver S9 fraction in the presence of acetyl-CoA, more than 50% of M1 (2 microM) was converted back to S4, but this conversion was not observed in the incubation with dog liver S9 fraction or human liver microsome. In vivo pharmacokinetic experiments showed that M1 could be rapidly converted back to S4 in rats, but a similar conversion was not observed in dogs. When S4 was administered, the formation of M1 was only observed in dogs due to the absence of NAT expression. Simultaneous fitting of the concentration time profiles of both S4 and M1 showed that more than 50% of S4 was deacetylated to M1 in dogs after i.v. administration of S4, whereas more than 80% of M1 was converted to S4 in rats after i.v. administration of M1. Considering the polymorphism in NAT expression, the interaction between M1 and NAT may raise concerns for drug-drug interactions during clinical applications of S4. The observed species differences suggested that interspecies scaling might not be applicable for predicting the metabolism and disposition of S4 in humans. PMID- 16272404 TI - Characterization of the in vitro metabolism of selective androgen receptor modulator using human, rat, and dog liver enzyme preparations. AB - Compound S4 [S-3-(4-acetylamino-phenoxy)-2-hydroxy-2-methyl-N-(4-nitro-3 trifluoromethyl-phenyl)-propionamide] is a novel nonsteroidal selective androgen receptor modulator that demonstrates tissue-selective androgenic and anabolic effects. The purpose of this in vitro study was to identify the phase I metabolites, potential species differences in metabolism, and the cytochromes P450 (P450s) involved in the phase I metabolism of S4 using 14C-S4, recombinant P450s, and other liver enzyme preparations from human, rat, and dog. The major phase I metabolism pathways of S4 in humans were identified as deacetylation of the B-ring acetamide group, hydrolysis of the amide bond, reduction of the A-ring nitro group, and oxidation of the aromatic rings, with deacetylation being the predominant pathway observed with most of the enzyme preparations tested. Among the major human P450 enzymes tested, CYP3A4 appeared to be one of the major phase I enzymes that could be responsible for the phase I metabolism of S4 [Km = 16.1 microM, Vmax = 1.6 pmol/(pmol x min)] in humans and mainly catalyzed the deacetylation, hydrolysis, and oxidation of S4. In humans, the cytosolic enzymes mainly catalyzed the hydrolysis reaction, whereas the microsomal enzymes primarily catalyzed the deacetylation reactions. Similar phase I metabolic profiles were observed in rats and dogs as well, except that the amide bond hydrolysis seemed to occur more rapidly in rats. In summary, these results showed that the major phase I reaction of S4 in human, rat, and dog is acetamide group deacetylation. PMID- 16272405 TI - Characterization of human cytochrome p450 enzymes involved in the metabolism of the piperidine-type phenothiazine neuroleptic thioridazine. AB - The aim of the present study was to identify human cytochrome P450 enzymes (P450s) involved in mono-2-, di-2-, and 5-sulfoxidation, and N-demethylation of the piperidine-type phenothiazine neuroleptic thioridazine in the human liver. The experiments were performed in vitro using cDNA-expressed human P450s (Supersomes 1A2, 2A6, 2B6, 2C9, 2C19, 2D6, 2E1, and 3A4), liver microsomes from different donors, and P450-selective inhibitors. The results indicate that CYP1A2 and CYP3A4 are the main enzymes responsible for 5-sulfoxidation and N demethylation (34-52%), whereas CYP2D6 is the basic enzyme that catalyzes mono-2- and di-2-sulfoxidation of thioridazine in human liver (49 and 64%, respectively). Besides CYP2D6, CYP3A4 contributes to a noticeable degree to thioridazine mono-2 sulfoxidation (22%). Therefore, the sulforidazine/mesoridazine ratio may be an additional and more specific marker than the mesoridazine/thioridazine ratio for assessing the activity of CYP2D6. In contrast to promazine and perazine, CYP2C19 insignificantly contributes to the N-demethylation of thioridazine. Considering serious side-effects of thioridazine and its 5-sulfoxide (cardiotoxicity), as well as strong dopaminergic D2 and noradrenergic alpha1 receptor-blocking properties of mono-2- and di-2-sulfoxides, the obtained results are of pharmacological and clinical importance, in particular, in a combined therapy. Knowledge of the catalysis of thioridazine metabolism helps to choose optimum conditions (a proper coadministered drug and dosage) to avoid undesirable drug interactions. PMID- 16272406 TI - Role of Mrp2 in the hepatic disposition of mycophenolic acid and its glucuronide metabolites: effect of cyclosporine. AB - Mycophenolic acid (MPA) is part of the immunosuppressant therapy for transplant recipients. This study examines the role of the canalicular transporter, Mrp2, and the effect of cyclosporin A (CsA), on the biliary secretion of the ether (MPAGe) and acyl (MPAGa) glucuronides of MPA. Isolated livers from Wistar rats (n = 6), or Wistar TR- rats (n = 6) were perfused with MPA (5 mg/l). A third group of Wistar rats (n = 6) was perfused with MPA and CsA (250 microg/l). There was no difference in the half-life, hepatic extraction ratio (E(H)), clearance or partial clearance of MPA to MPAGe, but there was a difference in partial clearance to MPAGa between control and CsA groups (0.9 +/- 0.4 versus 0.5 +/- 0.1 ml/min). TR- rats had a lower E(H) (0.59 +/- 0.30 versus 0.95 +/- 0.30), a lower clearance (18 +/- 8 versus 29 +/- 7 ml/min), and a longer half-life (19.5 +/- 10.3 versus 10.1 +/- 2.4 min) than controls. Compared to controls, MPAGe and MPAGa biliary excretion was reduced by 99% and 71.8%, respectively, in TR- rats, and 17.5% and 53.8%, respectively, in the MPA-CsA group. The biliary excretion of MPAGe is mediated by Mrp2, whereas that of MPAGa seems to depend on both Mrp2 and another unidentified canalicular transporter. Although CsA can inhibit Mrp2, our data suggest that it may also inhibit the hepatic glucuronidation of MPA in Wistar rats. PMID- 16272407 TI - Neuropathology in Drosophila membrane excitability mutants. AB - Mutations affecting ion channels and neuronal membrane excitability have been identified in Drosophila as well as in other organisms and characterized for their acute effects on behavior and neuronal function. However, the long-term effect of these perturbations on the maintenance of neuronal viability has not been studied in detail. Here we perform an initial survey of mutations affecting Na+ channels and K+ channels in Drosophila to investigate their effects on life span and neuronal viability as a function of age. We find that mutations that decrease membrane excitability as well as those that increase excitability can trigger neurodegeneration to varying degrees. Results of double-mutant interactions with dominant Na+/K+ ATPase mutations, which themselves cause severe neurodegeneration, suggest that excitotoxicity owing to hyperexcitability is insufficient to explain the resultant phenotype. Although the exact mechanisms remain unclear, our results suggest that there is an important link between maintenance of proper neuronal signaling and maintenance of long-term neuronal viability. Disruption of these signaling mechanisms in any of a variety of ways increases the incidence of neurodegeneration. PMID- 16272409 TI - Analyses of SUM1-1-mediated long-range repression. AB - In Saccharomyces cerevisiae, local repression is promoter specific and localized to a small region on the DNA, while silencing is promoter nonspecific, encompasses large domains of chromatin, and is stably inherited for multiple generations. Sum1p is a local repressor protein that mediates repression of meiosis-specific genes in mitotic cells while the Sir proteins are long-range repressors that stably silence genes at HML, HMR, and telomeres. The SUM1-1 mutation is a dominant neomorphic mutation that enables the mutant protein to be recruited to the HMR locus and repress genes, even in the absence of the Sir proteins. In this study we show that the mutation in Sum1-1p enabled it to spread, and the native HMR barrier blocked it from spreading. Thus, like the Sir proteins, Sum1-1p was a long-range repressor, but unlike the Sir proteins, Sum1 1p-mediated repression was more promoter specific, repressing certain genes better than others. Furthermore, repression mediated by Sum1-1p was not stably maintained or inherited and we therefore propose that Sum1-1p-mediated long-range repression is related but distinct from silencing. PMID- 16272408 TI - Diverse mitotic and interphase functions of condensins in Drosophila. AB - The condensin complex has been implicated in the higher-order organization of mitotic chromosomes in a host of model eukaryotes from yeasts to flies and vertebrates. Although chromosomes paradoxically appear to condense in condensin mutants, chromatids are not properly resolved, resulting in chromosome segregation defects during anaphase. We have examined the role of different condensin complex components in interphase chromatin function by examining the effects of various condensin mutations on position-effect variegation in Drosophila melanogaster. Surprisingly, most mutations affecting condensin proteins were often found to result in strong enhancement of variegation in contrast to what might be expected for proteins believed to compact the genome. This suggests either that the role of condensin proteins in interphase differs from their expected role in mitosis or that the way we envision condensin's activity needs to be modified to accommodate alternative possibilities. PMID- 16272410 TI - Genetic interactions between Nhp6 and Gcn5 with Mot1 and the Ccr4-Not complex that regulate binding of TATA-binding protein in Saccharomyces cerevisiae. AB - Our previous work suggests that the Nhp6 HMGB protein stimulates RNA polymerase II transcription via the TATA-binding protein TBP and that Nhp6 functions in the same functional pathway as the Gcn5 histone acetyltransferase. In this report we examine the genetic relationship between Nhp6 and Gcn5 with the Mot1 and Ccr4-Not complexes, both of which have been implicated in regulating DNA binding by TBP. We find that combining either a nhp6ab or a gcn5 mutation with mot1, ccr4, not4, or not5 mutations results in lethality. Combining spt15 point mutations (in TBP) with either mot1 or ccr4 also results in either a growth defect or lethality. Several of these synthetic lethalities can be suppressed by overexpression of TFIIA, TBP, or Nhp6, suggesting that these genes facilitate formation of the TBP TFIIA-DNA complex. The growth defect of a not5 mutant can be suppressed by a mot1 mutant. HO gene expression is reduced by nhp6ab, gcn5, or mot1 mutations, and the additive decreases in HO mRNA levels in nhp6ab mot1 and gcn5 mot1 strains suggest different modes of action. Chromatin immunoprecipitation experiments show decreased binding of TBP to promoters in mot1 mutants and a further decrease when combined with either nhp6ab or gcn5 mutations. PMID- 16272411 TI - Presynaptic UNC-31 (CAPS) is required to activate the G alpha(s) pathway of the Caenorhabditis elegans synaptic signaling network. AB - C. elegans mutants lacking the dense-core vesicle priming protein UNC-31 (CAPS) share highly similar phenotypes with mutants lacking a neuronal G alpha(s) pathway, including strong paralysis despite exhibiting near normal levels of steady-state acetylcholine release as indicated by drug sensitivity assays. Our genetic analysis shows that UNC-31 and neuronal G alpha(s) are different parts of the same pathway and that the UNC-31/G alpha(s) pathway is functionally distinct from the presynaptic G alpha(q) pathway with which it interacts. UNC-31 acts upstream of G alpha(s) because mutations that activate the G alpha(s) pathway confer similar levels of strongly hyperactive, coordinated locomotion in both unc 31 null and (+) backgrounds. Using cell-specific promoters, we show that both UNC 31 and the G alpha(s) pathway function in cholinergic motor neurons to regulate locomotion rate. Using immunostaining we show that UNC-31 is often concentrated at or near active zones of cholinergic motor neuron synapses. Our data suggest that presynaptic UNC-31 activity, likely acting via dense-core vesicle exocytosis, is required to locally activate the neuronal G alpha(s) pathway near synaptic active zones. PMID- 16272412 TI - Dormancy genes from weedy rice respond divergently to seed development environments. AB - Genes interacting with seed developmental environments control primary dormancy. To understand how a multigenic system evolved to adapt to the changing environments in weedy rice, we evaluated genetic components of three dormancy QTL in a synchronized nondormant genetic background. Two genetically identical populations segregating for qSD1, qSD7-1, and qSD12 were grown under greenhouse and natural conditions differing in temperature, relative humidity, and light intensity during seed development. Low temperatures tended to enhance dormancy in both conditions. However, genotypes responded to the environments divergently so that two populations displayed similar distributions for germination. Additive and/or dominance effects of the three loci explained approximately 90% of genetic variances and their epistases accounted for the remainder in each environment. The qSD1 and qSD7-1 main effects were increased, while the qSD12 additive effect was decreased by relatively low temperatures. Both gene main and epistatic effects were involved in G x E interactions, which in magnitude were greater than environmental main effect. The divergent responses of dormancy genes observed in this simple multigenic system presumably have selective advantages in natural populations adapted to changing environments and hence represent a genetic mechanism stabilizing the dormancy level of weedy rice ripened in different seasons or temperature regimes. PMID- 16272413 TI - The role of the N-terminal oligopeptide repeats of the yeast Sup35 prion protein in propagation and transmission of prion variants. AB - The cytoplasmic [PSI+] determinant of Saccharomyces cerevisiae is the prion form of the Sup35 protein. Oligopeptide repeats within the Sup35 N-terminal domain (PrD) presumably are required for the stable [PSI+] inheritance that in turn involves fragmentation of Sup35 polymers by the chaperone Hsp104. The nonsense suppressor [PSI+] phenotype can vary in efficiency probably due to different inheritable Sup35 polymer structures. Here we study the ability of Sup35 mutants with various deletions of the oligopeptide repeats to support [PSI+] propagation. We define the minimal region of the Sup35-PrD necessary to support [PSI+] as amino acids 1-64, which include the first two repeats, although a longer fragment, 1-83, is required to maintain weak [PSI+] variants. Replacement of wild type Sup35 with deletion mutants decreases the strength of the [PSI+] phenotype. However, with one exception, reintroducing the wild-type Sup35 restores the original phenotype. Thus, the specific prion fold defining the [PSI+] variant can be preserved by the mutant Sup35 protein despite the change of phenotype. Coexpression of wild-type and mutant Sup35 containing three, two, one, or no oligopeptide repeats causes variant-specific [PSI+] elimination. These data suggest that [PSI+] variability is primarily defined by differential folding of the Sup35-PrD oligopeptide-repeat region. PMID- 16272416 TI - A logistic regression mixture model for interval mapping of genetic trait loci affecting binary phenotypes. AB - Often in genetic research, presence or absence of a disease is affected by not only the trait locus genotypes but also some covariates. The finite logistic regression mixture models and the methods under the models are developed for detection of a binary trait locus (BTL) through an interval-mapping procedure. The maximum-likelihood estimates (MLEs) of the logistic regression parameters are asymptotically unbiased. The null asymptotic distributions of the likelihood ratio test (LRT) statistics for detection of a BTL are found to be given by the supremum of a chi2-process. The limiting null distributions are free of the null model parameters and are determined explicitly through only four (backcross case) or nine (intercross case) independent standard normal random variables. Therefore a threshold for detecting a BTL in a flanking marker interval can be approximated easily by using a Monte Carlo method. It is pointed out that use of a threshold incorrectly determined by reading off a chi2-probability table can result in an excessive false BTL detection rate much more severely than many researchers might anticipate. Simulation results show that the BTL detection procedures based on the thresholds determined by the limiting distributions perform quite well when the sample sizes are moderately large. PMID- 16272414 TI - Quantitative trait loci with age-specific effects on fecundity in Drosophila melanogaster. AB - Life-history theory and evolutionary theories of aging assume the existence of alleles with age-specific effects on fitness. While various studies have documented age-related changes in the genetic contribution to variation in fitness components, we know very little about the underlying genetic architecture of such changes. We used a set of recombinant inbred lines to map and characterize the effects of quantitative trait loci (QTL) affecting fecundity of Drosophila melanogaster females at 1 and 4 weeks of age. We identified one QTL on the second chromosome and one or two QTL affecting fecundity on the third chromosome, but these QTL affected fecundity only at 1 week of age. There was more genetic variation for fecundity at 4 weeks of age than at 1 week of age and there was no genetic correlation between early and late-age fecundity. These results suggest that different loci contribute to the variation in fecundity as the organism ages. Our data provide support for the mutation accumulation theory of aging as applied to reproductive senescence. Comparing the results from this study with our previous work on life-span QTL, we also find evidence that antagonistic pleiotropy may contribute to the genetic basis of senescence in these lines as well. PMID- 16272417 TI - Fixation of the human-specific CMP-N-acetylneuraminic acid hydroxylase pseudogene and implications of haplotype diversity for human evolution. AB - The human CMP-N-acetylneuraminic acid hydroxylase gene (CMAH) suffered deletion of an exon that encodes an active center for the enzyme approximately 3.2 million years ago (MYA). We analyzed a 7.3-kb intronic region of 132 CMAH genes to explore the fixation process of this pseudogene and the demographic implication of its haplotype diversity. Fifty-six variable sites were sorted into 18 different haplotypes with significant linkage disequilibrium. Despite the rather low nucleotide diversity, the most recent common ancestor at CMAH dates to 2.9 MYA. This deep genealogy follows shortly after the original exon deletion, indicating that the deletion has fixed in the population, although whether this fixation was facilitated by natural selection remains to be resolved. Remarkable features are exceptionally long persistence of two lineages and the confinement of one lineage in Africa, implying that some African local populations were in relative isolation while others were directly involved in multiple African exoduses of the genus Homo. Importantly, haplotypes found in Eurasia suggest interbreeding between then-contemporaneous human species. Although population structure within Africa complicates the interpretation of phylogeographic information of haplotypes, the data support a single origin of modern humans, but not with complete replacement of archaic inhabitants by modern humans. PMID- 16272419 TI - Genetic basis of drought resistance at reproductive stage in rice: separation of drought tolerance from drought avoidance. AB - Drought tolerance (DT) and drought avoidance (DA) are two major mechanisms in drought resistance of higher plants. In this study, the genetic bases of DT and DA at reproductive stage in rice were analyzed using a recombinant inbred line population from a cross between an indica lowland and a tropical japonica upland cultivar. The plants were grown individually in PVC pipes and two cycles of drought stress were applied to individual plants with unstressed plants as the control. A total of 21 traits measuring fitness, yield, and the root system were investigated. Little correlation of relative yield traits with potential yield, plant size, and root traits was detected, suggesting that DT and DA were well separated in the experiment. A genetic linkage map consisting of 245 SSR markers was constructed for mapping QTL for these traits. A total of 27 QTL were resolved for 7 traits of relative performance of fitness and yield, 36 QTL for 5 root traits under control, and 38 for 7 root traits under drought stress conditions, suggesting the complexity of the genetic bases of both DT and DA. Only a small portion of QTL for fitness- and yield-related traits overlapped with QTL for root traits, indicating that DT and DA had distinct genetic bases. PMID- 16272420 TI - Comparison of mail and telephone in assessing patient experiences in receiving care from medical group practices. AB - The medical group survey from the CAHPS (formerly Consumer Assessment of Health Plans Study) project, G-CAHPS, focuses on patient experiences in receiving care from their medical group practice. We compared mail and telephone responses to the G-CAHPS survey in a sample of 880 patients from four physician groups. Patients were randomly assigned to mode. Analyses included comparison of response rates, missing data, internal consistency reliability of six multi-item scales, and mean scores. A total of 537 phone completes and 343 mail completes were obtained (54% response rate). There were no significant differences in internal consistency by mode. In addition, there was only one significant mode difference in item and composite means by mode of administration after adjusting for case mix differences. This study indicates that mail and telephone modes of data collection for the G-CAHPS survey produce similar results. PMID- 16272415 TI - Nucleotide polymorphism and linkage disequilibrium in wild populations of the partial selfer Caenorhabditis elegans. AB - An understanding of the relative contributions of different evolutionary forces on an organism's genome requires an accurate description of the patterns of genetic variation within and between natural populations. To this end, I report a survey of nucleotide polymorphism in six loci from 118 strains of the nematode Caenorhabditis elegans. These strains derive from wild populations of several regions within France, Germany, and new localities in Scotland, in addition to stock center isolates. Overall levels of silent-site diversity are low within and between populations of this self-fertile species, averaging 0.2% in European samples and 0.3% worldwide. Population structure is present despite a lack of association of sequences with geography, and migration appears to occur at all geographic scales. Linkage disequilibrium is extensive in the C. elegans genome, extending even between chromosomes. Nevertheless, recombination is clearly present in the pattern of polymorphisms, indicating that outcrossing is an infrequent, but important, feature in this species ancestry. The range of outcrossing rates consistent with the data is inferred from linkage disequilibrium, using "scattered" samples representing the collecting phase of the coalescent process in a subdivided population. I propose that genetic variation in this species is shaped largely by population subdivision due to self fertilization coupled with long- and short-range migration between subpopulations. PMID- 16272421 TI - Are continuity clinic patients less satisfied when residents have a heavy inpatient workload? AB - The purpose of this study was to assess the influence of resident non-clinic workload on the satisfaction of continuity clinic patients. Over a 2-month period in 2002, residents and patients were surveyed at the University of Kentucky internal medicine continuity clinic. Residents provided a self-report of their non-clinic workload as light or medium versus heavy or extremely heavy. Patient satisfaction was assessed with a 7-item, 10-point scale with items derived from commonly used patient satisfaction instruments. In 168 patient encounters, patients were significantly less satisfied with their clinic visit if they were seen by a resident who had a heavier workload. In addition, these patients gave significantly lower ratings with regard to the amount of time spent with the patient during the visit, and how well the resident listened and paid attention. Although alternative explanations exist, we propose that heavy hospital workload is associated with decreased patient satisfaction in resident continuity clinic. PMID- 16272422 TI - Job satisfaction of physicians with congruent versus incongruent specialty choice. AB - Choosing a specialty within the occupation of medicine is problematic given the number of available specialty choices. A need exists for the development and modification of inventories to be used to help students with selecting a medical specialty. Furthermore, ratings of job satisfaction, regardless of specialty, are mixed with some physicians regretting choosing medicine as a career. Despite its use in medical specialty counseling, research is lacking regarding the Medical Specialty Preference Inventory (MSPI). We examined the predictive validity of the MSPI and compared job satisfaction of physicians (N=51) whose specialty was correctly (i.e., congruent) versus incorrectly (i.e., incongruent) predicted by the MSPI. The MSPI correctly predicted the specialty for 33%. Congruent physicians had higher job satisfaction than incongruent physicians. Results of this study may help individuals who provide medical specialty counseling or who are involved in designing tools and implementing programs to assist students with medical specialty decision making. PMID- 16272423 TI - Help on the line: telephone-triage use, outcomes, and satisfaction within an uninsured population. AB - Telephone triage programs have been shown to be cost-effective and favorably utilized by insured populations. However, there are 45 million Americans who are uninsured and who do not have access to telephone nursing. A telephone triage service was piloted for local uninsured residents. Within the 17-month trial period, 320 calls were received, representing 207 clients. This study reports on the results of the telephone survey with a cross-sectional sample of uninsured triage patrons (N = 80). One half reported they would have sought other medical care if the telephone triage service had not been available. Most callers (98%) believed that their health care concern was understood. Moreover, 98% agreed with the advice given, and 90% reported following up on the advice given. Overall satisfaction by the uninsured population with the telephone-based nurse triage service was positive and appears to be an effective and acceptable tool by those uninsured individuals who utilized its services. PMID- 16272424 TI - Achieving consensus on leadership competencies and outcome measures: The Pediatric Pulmonary Centers' experience. AB - Pediatric Pulmonary Centers (PPCs) are federally funded interdisciplinary leadership training programs aiming to improve the health of families and children. This article describes the process PPCs used to efficiently and effectively achieve consensus on leadership training competencies and outcome measures among a large and diverse group of health professionals. Phase 1 used a modified Delphi technique to develop an initial set of competencies and outcome measures. Phase 2 used the nominal group technique and modified focus group strategies to refine and prioritize the competencies and outcomes measures. Participants reported being highly satisfied with the process and outcomes. In Phase 3, a formal program evaluation instrument was implemented, designed to measure the competency and describe the career paths and leadership accomplishments of previous trainees. The consensus process adopted can serve as a model for academic and public health entities seeking to achieve consensus on program goals, strategies, methods, priorities, and outcomes. PMID- 16272418 TI - A cis-regulatory sequence within the yellow locus of Drosophila melanogaster required for normal male mating success. AB - Drosophila melanogaster males perform a courtship ritual consisting of a series of dependent fixed-action patterns. The yellow (y) gene is required for normal male courtship behavior and subsequent mating success. To better characterize the requirement for y in the manifestation of innate male sexual behavior, we measured the male mating success (MMS) of 12 hypomorphic y mutants and matched outbred-background controls using a y+ rescue element on a freely segregating minichromosome. We found that 4 hypomorphs significantly reduced MMS to varying degrees. Reduced MMS was largely independent of adult pigmentation patterns. These mutations defined a 300-bp regulatory region upstream of the transcription start, the mating-success regulatory sequence (MRS), whose function is required for normal MMS. Visualization of gene action via GFP and a Yellow antibody suggests that the MRS directs y transcription in a small number of cells in the third instar CNS, the developmental stage previously implicated in the role of y with regard to male courtship behavior. The presence of Yellow protein in these cells positively correlates with MMS in a subset of mutants. The MRS contains a regulatory sequence controlling larval pigmentation and a 35-bp sequence that is highly conserved within the genus Drosophila and is predicted to bind known transcription factors. PMID- 16272426 TI - Evaluation of a care coordination/home-telehealth program for veterans with diabetes: health services utilization and health-related quality of life. AB - We evaluated a Veterans Health Administration (VHA) care coordination/ home telehealth (CC/HT) program on the utilization of health care services and health related quality of life (HRQL) in veterans with diabetes. Administrative records of 445 veterans with diabetes were reviewed to compare health care service utilization in the 1-year period before and 1-year period post-enrollment and also examined self-reported HRQL at enrollment and 1 year later. Multivariate analyses indicated a statistically significant reduction in the proportion of patients who were hospitalized (50% reduction), emergency room use (11% reduction), reduction in the average number of bed days of care (decreased an average of 3.0 days), and improvement in the HRQL role-physical functioning, bodily pain, and social functioning. The results need to be interpreted with caution because we used a single-group study design that may be influenced by regression to the mean. Ideally, future research should use a randomized controlled trial design. PMID- 16272425 TI - Reliability and validity of the DCP among hispanic veterans. AB - The Diabetes Care Profile (DCP) was designed to measure psychosocial factors related to diabetes and its treatment. This study sought to determine the reliability and validity of the DCP in Hispanic veterans with Type 2 diabetes. Hispanic (n=81) and non-Hispanic White (n=238) patients were recruited at three southwestern VA hospitals. Scale reliabilities calculated by Cronbach's coefficient alpha revealed reliabilities ranging from .54 to .97 in Hispanics and .63 to .95 in non-Hispanic Whites. Only one scale, Monitoring Barriers, differed significantly between the two patient groups. Mean values on the DCP scales were consistent within and across ethnicities lending support for construct validity of the DCP in Hispanics. Convergent validity was also supported for DCP scales within the Hispanic patients as evidenced by correlations in expected directions with external measures. PMID- 16272427 TI - Readability level of health insurance portability and accountability act notices of privacy practices utilized by academic medical centers. AB - Recently enacted Health Insurance Portability and Accountability Act legislation requires health care professionals to provide patients with a "Notice of Privacy Practices" (NPP) document as part of their informed consent process to participate in assessment and treatment. We were interested in the readability of these forms. We attempted to find a NPP from an academic medical center in each state plus the District of Columbia where documents were placed on the institution's Web site. Documents were obtained from 40 jurisdictions and were then analyzed utilizing two measures of readability. The majority (65%) of these documents were written beyond the 12th grade reading level, and almost the entire sample (90%) fell in the difficult range of reading ease. Academic medical centers have an ethical obligation to improve the readability of these documents and should do so to improve clinical practice and reduce liability. PMID- 16272428 TI - Transcription and histone modifications in the recombination-free region spanning a rice centromere. AB - Centromeres are sites of spindle attachment for chromosome segregation. During meiosis, recombination is absent at centromeres and surrounding regions. To understand the molecular basis for recombination suppression, we have comprehensively annotated the 3.5-Mb region that spans a fully sequenced rice centromere. Although transcriptional analysis showed that the 750-kb CENH3 containing core is relatively deficient in genes, the recombination-free region differs little in gene density from flanking regions that recombine. Likewise, the density of transposable elements is similar between the recombination-free region and flanking regions. We also measured levels of histone H4 acetylation and histone H3 methylation at 176 genes within the 3.5-Mb span. Active genes showed enrichment of H4 acetylation and H3K4 dimethylation as expected, including genes within the core. Our inability to detect sequence or histone modification features that distinguish recombination-free regions from flanking regions that recombine suggest that recombination suppression is an epigenetic feature of centromeres maintained by the assembly of CENH3-containing nucleosomes within the core. CENH3-containing centrochromatin does not appear to be distinguished by a unique combination of H3 and H4 modifications. Rather, the varied distribution of histone modifications might reflect the composition and abundance of sequence elements that inhabit centromeric DNA. PMID- 16272430 TI - Suppression of the floral activator Hd3a is the principal cause of the night break effect in rice. AB - A short exposure to light in the middle of the night causes inhibition of flowering in short-day plants. This phenomenon is called night break (NB) and has been used extensively as a tool to study the photoperiodic control of flowering for many years. However, at the molecular level, very little is known about this phenomenon. In rice (Oryza sativa), 10 min of light exposure in the middle of a 14-h night caused a clear delay in flowering. A single NB strongly suppressed the mRNA of Hd3a, a homolog of Arabidopsis thaliana FLOWERING LOCUS T (FT), whereas the mRNAs of OsGI and Hd1 were not affected. The NB effect on Hd3a mRNA was maximal in the middle of the 14-h night. The phyB mutation abolished the NB effect on flowering and Hd3a mRNA, indicating that the NB effect was mediated by phytochrome B. Because expression of the other FT-like genes was very low and not appreciably affected by NB, our results strongly suggest that the suppression of Hd3a mRNA is the principal cause of the NB effect on flowering in rice. PMID- 16272429 TI - Arabidopsis nitric oxide synthase1 is targeted to mitochondria and protects against oxidative damage and dark-induced senescence. AB - The Arabidopsis thaliana protein nitric oxide synthase1 (NOS1) is needed for nitric oxide (NO) synthesis and signaling during defense responses, hormonal signaling, and flowering. The cellular localization of NOS1 was examined because it is predicted to be a mitochondrial protein. NOS1-green fluorescent protein fusions were localized by confocal microscopy to mitochondria in roots. Isolated mitochondria from leaves of wild-type plants supported Arg-stimulated NO synthesis that could be inhibited by NOS inhibitors and quenched by a NO scavenger; this NOS activity is absent in mitochondria isolated from nos1 mutant plants. Because mitochondria are a source of reactive oxygen species (ROS), which participate in senescence and programmed cell death, these parameters were examined in the nos1 mutant. Dark-induced senescence of detached leaves and intact plants progressed more rapidly in the mutant compared with the wild type. Hydrogen peroxide, superoxide anion, oxidized lipid, and oxidized protein levels were all higher in the mutant. These results demonstrate that NOS1 is a mitochondrial NOS that reduces ROS levels, mitigates oxidative damage, and acts as an antisenescence agent. PMID- 16272431 TI - Endoplasmic reticulum glucosidase II is required for pathogenicity of Ustilago maydis. AB - We identified a nonpathogenic strain of Ustilago maydis by tagging mutagenesis. The affected gene, glucosidase1 (gas1), displays similarity to catalytic alpha subunits of endoplasmic reticulum (ER) glucosidase II. We have shown that Gas1 localizes to the ER and complements the temperature-sensitive phenotype of a Saccharomyces cerevisiae mutant lacking ER glucosidase II. gas1 deletion mutants were normal in growth and mating but were more sensitive to calcofluor and tunicamycin. Mutant infection hyphae displayed significant alterations in the distribution of cell wall material and were able to form appressoria and penetrate the plant surface but arrested growth in the epidermal cell layer. Electron microscopy analysis revealed that the plant-fungal interface between mutant hyphae and the plant plasma membrane was altered compared with the interface of penetrating wild-type hyphae. This may indicate that gas1 mutants provoke a plant response. PMID- 16272432 TI - Identification and functional characterization of Arabidopsis PEROXIN4 and the interacting protein PEROXIN22. AB - Peroxins are genetically defined as proteins necessary for peroxisome biogenesis. By screening for reduced response to indole-3-butyric acid, which is metabolized to active auxin in peroxisomes, we isolated an Arabidopsis thaliana peroxin4 (pex4) mutant. This mutant displays sucrose-dependent seedling development and reduced lateral root production, characteristics of plant peroxisome malfunction. We used yeast two-hybrid analysis to determine that PEX4, an apparent ubiquitin conjugating enzyme, interacts with a previously unidentified Arabidopsis protein, PEX22. A pex4 pex22 double mutant enhanced pex4 defects, confirming that PEX22 is a peroxin. Expression of both Arabidopsis genes together complemented yeast pex4 or pex22 mutant defects, whereas expression of either gene individually failed to rescue the corresponding yeast mutant. Therefore, it is likely that the Arabidopsis proteins can function similarly to the yeast PEX4-PEX22 complex, with PEX4 ubiquitinating substrates and PEX22 tethering PEX4 to the peroxisome. However, the severe sucrose dependence of the pex4 pex22 mutant is not accompanied by correspondingly strong defects in peroxisomal matrix protein import, suggesting that this peroxin pair may have novel plant targets in addition to those important in fungi. Isocitrate lyase is stabilized in pex4 pex22, indicating that PEX4 and PEX22 may be important during the remodeling of peroxisome matrix contents as glyoxysomes transition to leaf peroxisomes. PMID- 16272434 TI - Formation kinetics of insulin-based amyloid gels and the effect of added metalloporphyrins. AB - The kinetics of insulin-based amyloid gel formation has been studied using extinction and fluorescence detection. The process is treated as autocatalytic, and the kinetic profiles are fit using a nonconventional analysis involving a time-dependent rate constant (factor): k(t) = k(o) + k(c)(k(c)t)(n). The dependence of the kinetic parameters on initial solution conditions of concentration, pH, and ionic strength has been investigated. A mechanism is proposed in which the rate-determining step involves the activation of insulin solute species into partially unfolded, structurally modified monomers, which then aggregate. The influence of added metalloporphyrins on the rate and extent of gel formation is described. Metal derivatives of tetrakis(4 sulfonatophenyl)porphine prove effective at inhibiting the aggregation of insulin via pathways that depend on concentration and identity of the incorporated metal. PMID- 16272435 TI - Effects of the number of actin-bound S1 and axial force on X-ray patterns of intact skeletal muscle. AB - Effects of the number of actin-bound S1 and of axial tension on x-ray patterns from tetanized, intact skeletal muscle fibers were investigated. The muscle relaxant, BDM, reduced tetanic M3 meridional x-ray reflection intensity (I(M3)), M3 spacing (d(M3)), and the equatorial I(11)/I(10) ratio in a manner consistent with a reduction in the fraction of S1 bound to actin rather than by generation of low-force S1-actin isomers. At complete force suppression, I(M3) was 78% of its relaxed value. BDM distorted dynamic I(M3) responses to sinusoidal length oscillations in a manner consistent with an increased cross-bridge contribution to total sarcomere compliance, rather than a changed S1 lever orientation in BDM. When the number of actin-bound S1 was varied by altering myofilament overlap, tetanic I(M3) at low overlap was similar to that in high [BDM] (79% of relaxed I(M3)). Tetanic d(M3) dependence on active tension in overlap experiments differed from that observed with BDM. At high BDM, tetanic d(M3) approached its relaxed value (14.34 nm), whereas tetanic d(M3) at low overlap was 14.50 nm, close to its value at full overlap (14.56 nm). This difference in tetanic d(M3) behavior was explicable by a nonlinear thick filament compliance which is extended by both active and passive tension. PMID- 16272433 TI - Arabidopsis fragile fiber8, which encodes a putative glucuronyltransferase, is essential for normal secondary wall synthesis. AB - Secondary walls in vessels and fibers of dicotyledonous plants are mainly composed of cellulose, xylan, and lignin. Although genes involved in biosynthesis of cellulose and lignin have been intensively studied, little is known about genes participating in xylan synthesis. We found that Arabidopsis thaliana fragile fiber8 (fra8) is defective in xylan synthesis. The fra8 mutation caused a dramatic reduction in fiber wall thickness and a decrease in stem strength. FRA8 was found to encode a member of glycosyltransferase family 47 and exhibits high sequence similarity to tobacco (Nicotiana plumbaginifolia) pectin glucuronyltransferase. FRA8 is expressed specifically in developing vessels and fiber cells, and FRA8 is targeted to Golgi. Comparative analyses of cell wall polysaccharide fractions from fra8 and wild-type stems showed that the xylan and cellulose contents are drastically reduced in fra8, whereas xyloglucan and pectin are elevated. Further structural analysis of cell walls revealed that although wild-type xylans contain both glucuronic acid and 4-O-methylglucuronic acid residues, xylans from fra8 retain only 4-O-methylglucuronic acid, indicating that the fra8 mutation results in a specific defect in the addition of glucuronic acid residues onto xylans. These findings suggest that FRA8 is a glucuronyltransferase involved in the biosynthesis of glucuronoxylan during secondary wall formation. PMID- 16272436 TI - Optical microscopy of growing insulin amyloid spherulites on surfaces in vitro. AB - Amyloid fibrils are often found arranged into large ordered spheroid structures, known as spherulites, occurring in vivo and in vitro. The spherulites are predominantly composed of radially ordered amyloid fibrils, which self-assemble from protein in solution. We have observed and measured amyloid spherulites forming from heat-treated solutions of bovine insulin at low pH. The spherulites form in large numbers as semispherical dome-shaped objects on the cell surfaces, showing that surface defects or impurities, or the substrates themselves, can provide good nucleation sites for their formation. Using optical microscopy, we have measured the growth of individual spherulites as a function of time and in various conditions. There is a lag time before nucleation of the spherulites. Once they have nucleated, they grow, each with a radius increasing linearly, or faster than linearly, with time. Remarkably, this growth period has a sudden end, at which all spherulites in the system suddenly stop growing. A model of spherulite formation based on the polymerization of oriented fibrils around a nucleus, from a precursor in solution, quantitatively accounts for the observed growth kinetics. Seeding of native insulin solutions with preformed spherulites led to the preformed spherulites growing without a lag time. This seeding behavior is evidence that the fibrils in the spherulites assemble from small protein species rather than fibrils. The density of the spherulites was also measured and found to be constant with respect to radius, indicating that the space fills as the spherulite grows. PMID- 16272437 TI - Field theoretic study of bilayer membrane fusion: II. Mechanism of a stalk-hole complex. AB - We use self-consistent field theory to determine structural and energetic properties of intermediates and transition states involved in bilayer membrane fusion. In particular, we extend our original calculations from those of the standard hemifusion mechanism, which was studied in detail in the first article of this series, to consider a possible alternative to it. This mechanism involves non-axial stalk expansion, in contrast to the axially symmetric evolution postulated in the classical mechanism. Elongation of the initial stalk facilitates the nucleation of holes and leads to destabilization of the fusing membranes via the formation of a stalk-hole complex. We study properties of this complex in detail, and show how transient leakage during fusion, previously predicted and recently observed in experiment, should vary with lipid architecture and tension. We also show that the barrier to fusion in the alternative mechanism is lower than that of the standard mechanism by a few k(B)T over most of the relevant region of system parameters, so that this alternative mechanism is a viable alternative to the standard pathway. We emphasize that any mechanism, such as this alternative one, which affects, even modestly, the line tension of a hole in a membrane, affects greatly the ability of that membrane to undergo fusion. PMID- 16272438 TI - Catch-bond model derived from allostery explains force-activated bacterial adhesion. AB - High shear enhances the adhesion of Escherichia coli bacteria binding to mannose coated surfaces via the adhesin FimH, raising the question as to whether FimH forms catch bonds that are stronger under tensile mechanical force. Here, we study the length of time that E. coli pause on mannosylated surfaces and report a double exponential decay in the duration of the pauses. This double exponential decay is unlike previous single molecule or whole cell data for other catch bonds, and indicates the existence of two distinct conformational states. We present a mathematical model, derived from the common notion of chemical allostery, which describes the lifetime of a catch bond in which mechanical force regulates the transitions between two conformational states that have different unbinding rates. The model explains these characteristics of the data: a double exponential decay, an increase in both the likelihood and lifetime of the high binding state with shear stress, and a biphasic effect of force on detachment rates. The model parameters estimated from the data are consistent with the force induced structural changes shown earlier in FimH. This strongly suggests that FimH forms allosteric catch bonds. The model advances our understanding of both catch bonds and the role of allostery in regulating protein activity. PMID- 16272439 TI - Exploring the complex folding kinetics of RNA hairpins: II. Effect of sequence, length, and misfolded states. AB - The complexity of RNA hairpin folding arises from the interplay between the loop formation, the disruption of the slow-breaking misfolded states, and the formation of the slow-forming native base stacks. We investigate the general physical mechanism for the dependence of the RNA hairpin folding kinetics on the sequence and the length of the hairpin loop and the helix stem. For example, 1), the folding would slow down when a stable GC basepair moves to the middle of the stem; 2), hairpin with GC basepair near the loop would fold/unfold faster than the one with GC near the tail of the stem; 3), within a certain range of the stem length, a longer stem can cause faster folding; and 4), certain misfolded states can assist folding through the formation of scaffold structures to lower the entropic barrier for the folding. All our findings are directly applicable and quantitatively testable in experiments. In addition, our results can be useful for molecular design to achieve desirable fast/slow-folding hairpins, hairpins with/without specific misfolded intermediates, and hairpins that fold along designed pathways. PMID- 16272440 TI - Exploring the complex folding kinetics of RNA hairpins: I. General folding kinetics analysis. AB - Depending on the nucleotide sequence, the temperature, and other conditions, RNA hairpin-folding kinetics can be very complex. The complexity with a wide range of cooperative and noncooperative kinetic behaviors arises from the interplay between the formation of the loops, the disruption of the misfolded states, and the formation of the rate-limiting base stacks. With a rate constant model and a kinetic-cluster theory, we explore the broad landscape for RNA hairpin-folding kinetics. The model is validated through direct tests against several experimental measurements. The general kinetic folding mechanisms and the predicted great variety of folding kinetics are directly applicable and quantitatively testable in experiments. The results from this study suggest that 1), previous experimental findings based on the individual hairpins revealed only a small fraction of much broader and more complex RNA hairpin-folding landscapes; 2), even for structures as simple as hairpins, universal folding timescales and pathways do not exist; and 3), to treat the loop size as the sole factor to determine the hairpin-folding rate is an oversimplification. PMID- 16272441 TI - Thymosin beta4 induces a conformational change in actin monomers. AB - Using fluorescence resonance energy transfer spectroscopy we demonstrate that thymosin beta(4) (tbeta(4)) binding induces spatial rearrangements within the small domain (subdomains 1 and 2) of actin monomers in solution. Tbeta(4) binding increases the distance between probes attached to Gln-41 and Cys-374 of actin by 2 A and decreases the distance between the purine base of bound ATP (epsilonATP) and Lys-61 by 1.9 A, whereas the distance between Cys-374 and Lys-61 is minimally affected. Distance determinations are consistent with tbeta(4) binding being coupled to a rotation of subdomain 2. By differential scanning calorimetry, tbeta(4) binding increases the cooperativity of ATP-actin monomer denaturation, consistent with conformational rearrangements in the tbeta(4)-actin complex. Changes in fluorescence resonance energy transfer are accompanied by marked reduction in solvent accessibility of the probe at Gln-41, suggesting it forms part of the binding interface. Tbeta(4) and cofilin compete for actin binding. Tbeta(4) concentrations that dissociate cofilin from actin do not dissociate the cofilin-DNase I-actin ternary complex, consistent with the DNase binding loop contributing to high-affinity tbeta(4)-binding. Our results favor a model where thymosin binding changes the average orientation of actin subdomain 2. The tbeta(4)-induced conformational change presumably accounts for the reduced rate of amide hydrogen exchange from actin monomers and may contribute to nucleotide dependent, high affinity binding. PMID- 16272442 TI - Antagonist-induced deadhesion of specifically adhered vesicles. AB - By use of a model system consisting of giant vesicles adhering to flat substrates, we identified, both experimentally and theoretically, two new control mechanisms for antagonist-induced deadhesion. Adhesion is established by specific binding of surface-grafted E-selectin and vesicle-carrying oligosaccharide Lewis(X). Deadhesion is achieved by controlled titration of monoclonal antibodies against E-selectin. The first mechanism is characterized by a considerable retraction of the contact zone resulting in a loss of contact area between the vesicle and the substrate. Within the developed theoretical framework, the observed equilibrium state is understood as a balance between the spreading pressure of the vesicle and the antagonist-induced lateral pressure at the edge of the contact zone. In the second mechanism, the antibodies induce unbinding by penetrating the contact zone without significantly affecting its size. This process reveals the decomposition of the adhesion zone into microdomains of tight binding separated by strongly fluctuating sections of the membrane. Both experiment and theory show a sigmoidal decrease of the number of bound ligands as a function of the logarithm of antagonist concentration. The work presented herein also provides a new method for the determination of the receptor binding affinity of either the surface-embedded ligands or the competing antagonist molecules. PMID- 16272443 TI - A protein molecule in a mixed solvent: the preferential binding parameter via the Kirkwood-Buff theory. PMID- 16272444 TI - Probing the effect of point mutations at protein-protein interfaces with free energy calculations. AB - We have studied the effect of point mutations of the primary binding residue (P1) at the protein-protein interface in complexes of chymotrypsin and elastase with the third domain of the turkey ovomucoid inhibitor and in trypsin with the bovine pancreatic trypsin inhibitor, using molecular dynamics simulations combined with the linear interaction energy (LIE) approach. A total of 56 mutants have been constructed and docked into their host proteins. The free energy of binding could be reliably calculated for 52 of these mutants that could unambiguously be fitted into the binding sites. We find that the predicted binding free energies are in very good agreement with experimental data with mean unsigned errors between 0.50 and 1.03 kcal/mol. It is also evident that the standard LIE model used to study small drug-like ligand binding to proteins is not suitable for protein-protein interactions. Three different LIE models were therefore tested for each of the series of protein-protein complexes included, and the best models for each system turn out to be very similar. The difference in parameterization between small drug-like compounds and protein point mutations is attributed to the preorganization of the binding surface. Our results clearly demonstrate the potential of free energy calculations for probing the effect of point mutations at protein-protein interfaces and for exploring the principles of specificity of hot spots at the interface. PMID- 16272445 TI - Conversion of a porin-like peptide channel into a gramicidin-like channel by glycine to D-alanine substitutions. AB - The beta-barrel and beta-helix formation, as in porins and gramicidin, respectively, represent two distinct mechanisms for ion channel formation by beta sheet proteins in membranes. The design of beta-barrel proteins is difficult due to incomplete understanding of the basic principles of folding. The design of gramicidin-like beta-helix relies on an alternating pattern of L- and D-amino acid sequences. Recently we noticed that a short beta-sheet peptide (xSxG)(6), can form porin-like channels via self-association in membranes. Here, we proposed that glycine to D-alanine substitutions of the N-formyl-(xSxG)(6) would transform the porin-like channel into a gramicidin-like beta(12)-helical channel. The requirement of an N-formyl group for channel activity, impermeability to cations with a diameter >4 A, high monovalent cation selectivity, and the absence of either voltage gating or subconductance states upon D-alanine substitution support the idea of a gramicidin-like channel. Moreover, the circular dichroism spectrum in membranes is different, indicating a change in regular beta-sheet backbone structure. The conversion of a complex porin-like channel into a gramicidin-like channel provides a link between two different mechanisms of beta sheet channel formation in membranes and emphasizes the importance of glycine and D-amino acid residues in protein folding and function and in the engineering of ion channels. PMID- 16272446 TI - Anionic phospholipid interactions with the potassium channel KcsA: simulation studies. AB - Molecular dynamics (MD) simulations have been used to unmask details of specific interactions of anionic phospholipids with intersubunit binding sites on the surface of the bacterial potassium channel KcsA. Crystallographic data on a diacyl glycerol fragment at this site were used to model phosphatidylethanolamine (PE), or phosphatidylglycerol (PG), or phosphatidic acid (PA) at the intersubunit binding sites. Each of these models of a KcsA-lipid complex was embedded in phosphatidyl choline bilayer and explored in a 20 ns MD simulation. H-bond analysis revealed that in terms of lipid-protein interactions PA > PG >> PE and revealed how anionic lipids (PG and PA) bind to a site provided by two key arginine residues (R(64) and R(89)) at the interface between adjacent subunits. A 27 ns simulation was performed in which KcsA (without any lipids initially modeled at the R(64)/R(89) sites) was embedded in a PE/PG bilayer. There was a progressive specific increase over the course of the simulation in the number of H-bonds of PG with KcsA. Furthermore, two specific PG binding events at R(64)/R(89) sites were observed. The phosphate oxygen atoms of bound PG formed H bonds to the guanidinium group of R(89), whereas the terminal glycerol H-bonded to R(64). Overall, this study suggests that simulations can help identify and characterize sites for specific lipid interactions on a membrane protein surface. PMID- 16272447 TI - Cholesterol depletion induces solid-like regions in the plasma membrane. AB - Glycosylphosphatidylinositol-linked and transmembrane major histocompatibility complex (MHC) class II I-E(k) proteins, as well as N-(6 tetramethylrhodaminethiocarbamoyl)-1,2-dihexadecanoyl-sn-glycero-3 phosphoethanolamine (Tritc-DHPE), are used as probes to determine the effect of cholesterol concentration on the organization of the plasma membrane at temperatures in the range 22 degrees C-42 degrees C. Cholesterol depletion caused a decrease in the diffusion coefficients for the MHC II proteins and also for a slow fraction of the Tritc-DHPE population. At 37 degrees C, reduction of the total cell cholesterol concentration results in a smaller suppression of the translational diffusion for I-E(k) proteins (twofold) than was observed in earlier work at 22 degrees C (five sevenfold) Vrljic, M., S. Y. Nishimura, W. E. Moerner, and H. M. McConnell. 2005. Biophys. J. 88:334-347. At 37 degrees C, the diffusion of both I-E(k) proteins is Brownian (0.9 < alpha-parameter < 1.1). More than 99% of the protein population diffuses homogeneously when imaged at 65 frames per s. As the temperature is raised from 22 degrees C to 42 degrees C, a change in activation energy is seen at approximately 35 degrees C in the Arrhenius plots. Cytoskeletal effects appear to be minimal. These results are consistent with a previously described model of solid-like domain formation in the plasma membrane. PMID- 16272448 TI - Response to acute lung infection with mucoid Pseudomonas aeruginosa in cystic fibrosis mice. AB - RATIONALE: Cystic fibrosis is caused by defects in the cystic fibrosis transmembrane conductance regulator gene, which codes for a chloride channel, but the role of this chloride channel in inflammation induced by lung infection with Pseudomonas aeruginosa remains to be defined. OBJECTIVES: We tested the hypothesis that loss of this chloride channel alone is sufficient to cause excessive inflammation in response to inflammatory stimuli. METHODS: We investigated the response of cystic fibrosis and wild-type mice to mucoid P. aeruginosa administered by insufflation. MEASUREMENTS: The host responses measured included survival, weight change, lung morphometry, bacterial clearance, and inflammatory mediators, and cell counts were assessed in bronchoalveolar lavage fluid. MAIN RESULTS: Depending on the dose administered and frequency of dosing, cystic fibrosis mice experienced significantly higher mortality rates, greater weight loss, higher lung pathology scores, and higher inflammatory mediator and neutrophil levels compared with wild-type mice, even after the bacteria had been cleared. Surprisingly, bacteria were cleared just as rapidly in cystic fibrosis mice as in wild-type mice, and sepsis was not observed. Chronic lung infections could not be established with mucoid P. aeruginosa in either cystic fibrosis or wild-type mice. CONCLUSIONS: Absence of this chloride channel alone appears sufficient for exaggerated inflammation and excess mortality compared with wild-type controls in the face of mucoid P. aeruginosa lung infection. To establish chronic infection, additional factors such as bacterial trapping or poor clearance may be required. PMID- 16272449 TI - Association between adrenal insufficiency and ventilator weaning. AB - RATIONALE: Adrenal insufficiency is a common disorder in critically ill patients with mechanical ventilation and is usually associated with higher mortality and poor clinical outcome. OBJECTIVES: To determine whether stress dose corticosteroid supplementation can improve ventilator weaning and clinical outcome in patients with adrenal insufficiency. METHODS: A prospective, randomized, placebo controlled, double-blinded study was conducted in the intensive care unit of a tertiary teaching hospital. A total of 93 mechanically ventilated patients were enrolled in the ventilator weaning trial. Adrenal function was assessed in all patients. Patients with adrenal insufficiency were randomized to the treatment group (50 mg intravenous hydrocortisone every 6 h) and the placebo group. MEASUREMENTS AND MAIN RESULTS: The successful ventilator weaning percentage was significantly higher in the adequate adrenal reserve group (88.4%) and in the stress dose hydrocortisone treatment group (91.4%) than in the placebo group (68.6%). The weaning period was shorter in the hydrocortisone treatment group than in the placebo group. No significant adverse effects were observed in the corticosteroid treatment group. CONCLUSIONS: For patients with respiratory failure, early identification of adrenal insufficiency and appropriate supplementation with stress dose hydrocortisone increase the success of ventilator weaning and shortens the weaning period. PMID- 16272450 TI - Impact of Burkholderia dolosa on lung function and survival in cystic fibrosis. AB - RATIONALE: Chronic infection with Burkholderia cepacia complex bacteria in cystic fibrosis is associated with accelerated decline in pulmonary function and increased mortality. Clinical implications of the recently characterized genomovar VI, B. dolosa, are unknown. OBJECTIVES: Characterization of impact of B. dolosa on pulmonary function and mortality in cystic fibrosis. METHODS: We compared patients chronically infected with B. dolosa (n = 31) with unmatched patients with B. multivorans (n = 24) and with age- and sex-matched control subjects without Burkholderia species (n = 58). We analyzed rates of pulmonary function decline (% predicted FEV(1)) using a random effects model assuming segmented linear trends. All available FEV(1) measurements from 5 yr (median, 4.8) before until 2.5 yr (median, 1.5) after the first positive culture for Burkholderia (reference date) were analyzed. Survival was compared using the Kaplan-Meier method and proportional hazards model. MEASUREMENTS AND MAIN RESULTS: Baseline FEV(1) and rate of decline were similar in the cohorts. Decline in FEV(1) after the reference date accelerated in patients with B. dolosa (-2.3 percentage points/yr pre vs. -7.1 post, p = 0.002), but was unchanged in the B. multivorans and control patients (-2.3 vs. -0.8 post, p = 0.38, and -2.1 pre vs. 0.5 post, p = 0.20, respectively). The probability of dying within 18 mo of the reference date was 13, 7, and 3% for B. dolosa, B. multivorans, and control patients, respectively (B. dolosa vs. control hazard ratio, 10.8; 95% confidence interval, 1.3-92.8; p = 0.03). CONCLUSIONS: B. dolosa chronic infection in cystic fibrosis is associated with accelerated loss of lung function and decreased survival. PMID- 16272452 TI - Provocative optimism in the treatment of early stage disease. PMID- 16272451 TI - Improvement of lung compliance during postnatal adaptation correlates with airway sodium transport. AB - RATIONALE: Fetal lung liquid secretion is coupled with chloride transport into the lung lumen. The postnatal clearance of lung liquid is dependent on osmotic force generated by active sodium absorption. OBJECTIVE: To study the interaction between airway epithelial sodium transport and postnatal lung function. METHODS: We determined lung compliance and nasal transepithelial potential difference as a measure of airway ion transport and epithelial sodium channel gene expression in 41 healthy newborn infants during the first 50 h after birth. MEASUREMENTS AND MAIN RESULTS: Lung compliance improved significantly during the study period, whereas nasal potential difference remained constant. There was a significant decrease in the expressions of beta and gamma subunits of the epithelial sodium channel. A positive correlation existed between amiloride-sensitive nasal potential difference measured at 1-4 h of age and lung compliance at 21-27 h of age. We found no correlation between the molecular data and functional measurements. CONCLUSIONS: An important part of pulmonary adaptation takes place during the first hour after birth. The improvement of lung compliance continues over the first postnatal days and coincides with down-regulation of epithelial sodium channel beta and gamma subunit expression. PMID- 16272453 TI - Rapid evolution in colorectal cancer: therapy now and over the next five years. AB - A large number of patients with colorectal cancer have relatively early disease, and thus, adjuvant therapy has the potential to save lives. In stage III patients, there has been a steady improvement in 3-year disease-free survival with the use of 5-fluorouracil/leucovorin (5-FU/LV) regimens and capecitabine (Xeloda); Hoffmann-La Roche Inc., Nutley, NJ, http://www.rocheusa.com) regimens. A median survival longer than 20 months was observed in patients with metastatic disease when treated with combination chemotherapy containing oxaliplatin (Eloxatin); Sanofi-Synthelabo Inc., New York, http://www.sanofi-synthelabo.us) or irinotecan (Camptosar); Pfizer Pharmaceuticals, New York, http://www.pfizer.com). This has led to 5-FU/LV/oxaliplatin becoming standard therapy, along with 5 FU/LV/irinotecan. New data confirm the beneficial effect on disease-free survival of adding oxaliplatin to adjuvant colorectal cancer regimens based on 5-FU. These regimens show an effect when given in bolus as well as in infusional schedules. Interest in future adjuvant regimens focuses on the potential additional benefit of molecularly targeted agents, such as bevacizumab (Avastin); Genentech, Inc., South San Francisco, CA, http://www.gene.com), and on the ability of applied genomics to distinguish between high- and low-risk populations. PMID- 16272454 TI - Can we approach zero relapse in breast cancer? AB - Adjuvant hormonal therapy and adjuvant chemotherapy have contributed significantly to the falling rates of breast cancer mortality. The introduction of taxanes and aromatase inhibitors in the adjuvant setting represents recent important improvements. More recently, the demonstration of significant benefit in the adjuvant setting with novel molecular targeted therapies (such as trastuzumab [Herceptin; Genentech, Inc., South San Francisco, CA, http://www.gene.com]) is already beginning to have a substantial impact on the adjuvant treatment of patients with certain tumor characteristics (i.e., HER-2 positivity). Neoadjuvant treatment represents an approach that offers an intermediate end point (i.e., pathologic complete response) that can be used as a marker of therapeutic activity. Furthermore, the use of genomic profiling is starting to replace the traditional prognostic and predictive factors currently used to estimate risks for recurrence and response to particular adjuvant therapies. These recent developments have demonstrated that the notion of approaching zero relapse in breast cancer patients is now within our reach. PMID- 16272455 TI - High-risk localized prostate cancer: integrating chemotherapy. AB - Docetaxel (Taxotere); Aventis Pharmaceuticals Inc., Bridgewater, NJ, http://www.aventispharma-us.com) is the first agent to significantly extend survival in hormone-refractory prostate cancer. Because agents active in advanced cancers tend to be beneficial in earlier stage disease, docetaxel is now to be assessed, along with hormonal therapy, in the adjuvant setting among patients whose localized prostate cancer has features that put them at particular risk for recurrence and cancer-specific mortality. Data from a pilot study suggest that neo-adjuvant treatment with docetaxel may be appropriate for selected high-risk patients and that such treatment can be undertaken without increasing surgical morbidity. Gene-expression profiling of tissue before and after docetaxel treatment is providing further insight into its effects. A randomized trial, conducted by the Cancer and Leukemia Group B, will evaluate neoadjuvant docetaxel in high-risk patients, whereby patients will be randomized to either immediate radical prostatectomy or surgery preceded by hormonal therapy plus docetaxel. Another large randomized trial will be evaluating the effect of adjuvant hormonal therapy with or without docetaxel in high-risk men after radical prostatectomy. PMID- 16272456 TI - How today's developments in the treatment of non-small cell lung cancer will change tomorrow's standards of care. AB - Cisplatin (Platinol; Bristol-Myers Squibb, Princeton, NJ, http://www.bms.com) and carboplatin (Paraplatin; Bristol-Myers Squibb), together with newer chemotherapies, such as docetaxel (Taxotere; Aventis Pharmaceuticals Inc., Bridgewater, NJ, http://www.aventispharma-us.com), paclitaxel (Taxol; Bristol Myers Squibb), vinorelbine (Navelbine; GlaxoSmith-Kline, Philadelphia, http://www.gsk.com), pemetrexed (Alimta; Eli Lilly and Company, Indianapolis, http://www.lilly.com), and gemcitabine (Gemzar; Eli Lilly and Company), have improved treatment outcomes in both advanced non-small cell lung cancer (NSCLC) and in the adjuvant/neoadjuvant setting. Newer systemic treatments for NSCLC, used in advanced stage IV management, are beginning to be studied in earlier stages of the disease, when treatment is better tolerated and potentially curative. Hopefully, newer agents with proven efficacies in advanced disease will enhance curability. Following the successful addition of bevacizumab (Avastin; Genentech, Inc., South San Francisco, CA, http://www.gene.com) to carboplatin/paclitaxel in advanced disease, bevacizumab is now being incorporated into adjuvant and neoadjuvant trials. Trials in stage IB-IIIA patients will study neoadjuvant docetaxel/cisplatin/bevacizumab. The discovery that patients with exon 19 and 21 mutations in the epidermal growth factor receptor gene EGFR have around an 80% response rate to gefitinib (Iressa; AstraZeneca Pharmaceuticals, Wilmington, DE, http:// www.astrazeneca-us.com) and that this response confers survival benefit indicates its potential utility for mutation-positive patients with advanced- and earlier-stage disease. Clinical characteristics, such as never smoking status and adenocarcinoma, and especially bronchioloalveolar carcinoma histological features, can also identify individuals likely to respond to EGFR tyrosine kinase inhibitors. Studies of neoadjuvant erlotinib (Tarceva; OSI Pharmaceuticals, Inc., Melville, NY, http://www.osip.com) in operable NSCLC are planned. One such study includes cisplatin and docetaxel. Effective development of active agents and disease management based on molecular profiling of lung tumors will change tomorrow's standard of care. PMID- 16272457 TI - Gene-expression profiling and the future of adjuvant therapy. AB - Gene-expression profiling can help distinguish between patients at high risk and those at low risk for developing distant metastases, and so identify patients for adjuvant therapy. For several years, the Netherlands Cancer Institute has been working on gene-expression profiling of breast cancer using a microarray platform containing 25,000 genes. Using supervised classification, a prognostic classifier consisting of 70 genes could be identified. In addition to providing prognostic information, gene profiling should also enable us to detect patients who are likely to respond to particular adjuvant interventions. Well-known predictors for response to systemic therapy include estrogen receptor status HER-2 status, c-kit mutation, and epidermal growth factor receptor mutation. Because of the long periods required for predicting responsiveness in the adjuvant setting, neoadjuvant trials promise far quicker results. Several neoadjuvant studies are under way or planned to investigate gene-expression profiling as a means of predicting the therapeutic response to docetaxel (Taxotere; Aventis Pharmaceuticals Inc., Bridgewater, NJ, http://www.aventispharma-us.com), paclitaxel (Taxol; Bristol-Myers Squibb, Princeton, NJ, http://www.bms.com), cyclophosphamide, and doxorubicin (Adriamycin; Bedford Laboratories, Bedford, OH, http://www.bedfordlabs.com) in breast cancer patients. It is expected that in the coming years an increasing number of novel prognostic and predictive tests will help in guiding the adjuvant systemic treatment of breast cancer and other malignancies. PMID- 16272458 TI - In vivo particle uptake by airway macrophages in healthy volunteers. AB - We combined two techniques, radiolabeled aerosol inhalation delivery and induced sputum, to examine in vivo the time course of particle uptake by airway macrophages in 10 healthy volunteers. On three separate visits, induced sputum was obtained 40, 100, and 160 min after inhalation of radiolabeled sulfur colloid (SC) aerosol (Tc99 m-SC, 0.2 microm colloid size delivered in 6-microm droplets). On a fourth visit (control) with no SC inhalation, induced sputum was obtained and SC particles were incubated (37 degrees C) in vitro with sputum cells for 40, 100, and 160 min (matching the times associated with in vivo sampling). Total and differential cell counts were recorded for each sputum sample. Compared with 40 min (6 +/- 3%), uptake in vivo was significantly elevated at 100 (31 +/- 5%) and 160 min (27 +/- 4%); both were strongly associated with the number of airway macrophages (R = 0.8 and 0.7, respectively); and the number and proportion of macrophages at 40 min were significantly (P < 0.05) elevated compared with control (1,248 +/- 256 versus 555 +/- 114 cells/mg; 76 +/- 6% versus 60 +/- 5%). Uptake in vitro increased in a linear fashion over time and was maximal at 160 min (40 min, 12 +/- 2%; 100 min, 16 +/- 4%; 160 min, 24 +/- 6%). These data suggest that airway surface macrophages in healthy subjects rapidly engulf insoluble particles. Further, macrophage recruitment and phagocytosis-modifying agents are factors in vivo that likely affect particle uptake and its time course. PMID- 16272459 TI - Beta-catenin in the fibroproliferative response to acute lung injury. AB - Resolution of alveolar epithelial/capillary membrane damage after acute lung injury requires coordinated and effective tissue repair to reestablish a functional alveolar epithelial/capillary membrane barrier. We hypothesized that signaling pathways important in lung alveolar bud ontogeny are activated in the recovery and remodeling phases after profound oxidant stress lung injury in a murine model. To test this, we characterized the expression of noncanonical beta catenin pathway proteins E-cadherin, integrin-linked kinase-1, and beta-catenin in mice undergoing normoxic recovery after exposure to butylated hydroxytoluene (BHT, ionol) and concomitant sublethal (75% O2) hyperoxia. Mice developed early acute lung injury with subsequent inflammation, collagen deposition, interstitial cellular proliferation, and lung architectural distortion. Reduced E-cadherin expression after 6 d of BHT and hyperoxia was accompanied by enhanced expression and nuclear localization of beta-catenin and increased integrin-linked kinase-1 expression during subsequent normoxic recovery. This resulted in increased expression of the cotranscriptional regulators TCF-1 and -3 and cyclin D1. Proliferation of murine lung epithelial-12 cells in vitro after 8 h of treatment with BHT quinone-methide and hyperoxia and 48 h of normoxic recovery was enhanced 2.7-fold compared with vehicle-treated control mice at the same time point. BHT/hyperoxia-exposed mice treated with the pan-caspase inhibitor z-ASP had increased acute lung injury and reduced survival despite the presence of TUNEL positive cells, suggesting enhanced lung cell necrosis. Beta-catenin expression was reduced in z-ASP-co-treated lungs after BHT/hyperoxia. The noncanonical cadherin-beta-catenin axis is associated with fibroproliferative repair after BHT/hyperoxia exposure and may regulate epithelial proliferation and lung matrix remodeling and repair in response to lung injury. PMID- 16272461 TI - Oxidant generation promotes iron sequestration in BEAS-2B cells exposed to asbestos. AB - Lung injury after asbestos exposure is associated with an oxidative stress that is catalyzed by iron in the fiber matrix, complexed to the surface, or both. We tested the hypothesis that the cellular response to asbestos includes the transport and sequestration of this iron through (1) generation of superoxide for ferrireduction, (2) up-regulation of divalent metal transporter-1 (DMT1) for intracellular transport of Fe2+, and (3) increased production of cellular ferritin where the metal is stored in a catalytically less reactive state. BEAS 2B cells with normal and elevated Cu,Zn superoxide dismutase (SOD) expression were employed for in vitro investigations. After exposure of these cells to asbestos, we demonstrated by fluorescence methodology a significantly increased generation of SOD with ferrireductive capacity. Fiber exposure also increased DMT1 protein and mRNA expression in the BEAS-2B cells. Incubation with asbestos elevated cellular iron and ferritin concentrations, and these responses were diminished in cells with an enhanced expression of SOD. Finally, fiber exposure increased supernatant concentrations of interleukin 8, but this inflammatory mediator was actually increased in cells with elevated SOD expression. We conclude that the response of respiratory epithelial cells to asbestos includes oxidant-mediated mechanisms to sequester catalytically active iron associated with the fiber. PMID- 16272460 TI - TNF-alpha sensitizes normal and fibrotic human lung fibroblasts to Fas-induced apoptosis. AB - Pulmonary accumulation of fibroblasts and myofibroblasts in idiopathic pulmonary fibrosis/usual interstitial pneumonia (IFP/UIP) has been linked to (1) increased migration of a circulating pool of fibrocytes, (2) cell proliferation, and (3) resistance to apoptosis. The mechanism of physiologic apoptosis of lung fibroblasts is poorly understood. Using normal and fibrotic human lung fibroblasts and the human lung fibroblast cell line, MRC-5, we examined the regulation of Fas-induced apoptosis by the proinflammatory cytokines TNF-alpha and IFN-gamma. Herein, we show that the basal resistance of lung fibroblasts and myofibroblasts to Fas-induced apoptosis is overcome by sensitization with TNF alpha. IFN-gamma did not sensitize cells to Fas-induced apoptosis, but exhibited synergistic activity with TNF-alpha. Sensitization by TNF-alpha was observed in MRC-5 cells and in fibroblasts and myofibroblasts from normal and fibrotic human lung, suggesting that this represents a conserved mechanism to engage Fas-induced apoptosis. The mechanism of sensitization was localized at the level of recruitment of the adapter protein, FADD, to the cytoplasmic domain of Fas. Collectively, these findings suggest that fibroblast apoptosis involves two steps, sensitization and induction, and that inadequate pulmonary inflammation in IPF/UIP may favor fibroblast accumulation by reducing sensitization to apoptosis. PMID- 16272462 TI - Protocols for the assurance of microarray data quality and process control. AB - Microarrays represent a powerful technology that provides the ability to simultaneously measure the expression of thousands of genes. However, it is a multi-step process with numerous potential sources of variation that can compromise data analysis and interpretation if left uncontrolled, necessitating the development of quality control protocols to ensure assay consistency and high quality data. In response to emerging standards, such as the minimum information about a microarray experiment standard, tools are required to ascertain the quality and reproducibility of results within and across studies. To this end, an intralaboratory quality control protocol for two color, spotted microarrays was developed using cDNA microarrays from in vivo and in vitro dose-response and time course studies. The protocol combines: (i) diagnostic plots monitoring the degree of feature saturation, global feature and background intensities, and feature misalignments with (ii) plots monitoring the intensity distributions within arrays with (iii) a support vector machine (SVM) model. The protocol is applicable to any laboratory with sufficient datasets to establish historical high- and low-quality data. PMID- 16272463 TI - Use of biochemical kinetic data to determine strain relatedness among Salmonella enterica subsp. enterica isolates. AB - Classical biotyping characterizes strains by creating biotype profiles that consider only positive and negative results for a predefined set of biochemical tests. This method allows Salmonella subspecies to be distinguished but does not allow serotypes and phage types to be distinguished. The objective of this study was to determine the relatedness of isolates belonging to distinct Salmonella enterica subsp. enterica serotypes by using a refined biotyping process that considers the kinetics at which biochemical reactions take place. Using a Vitek GNI+ card for the identification of gram-negative organisms, we determined the biochemical kinetic reactions (28 biochemical tests) of 135 Salmonella enterica subsp. enterica strains of pig origin collected in Spain from 1997 to 2002 (59 Salmonella serotype Typhimurium strains, 25 Salmonella serotype Typhimurium monophasic variant strains, 25 Salmonella serotype Anatum strains, 12 Salmonella serotype Tilburg strains, 7 Salmonella serotype Virchow strains, 6 Salmonella serotype Choleraesuis strains, and 1 Salmonella enterica serotype 4,5,12:-:- strain). The results were expressed as the colorimetric and turbidimetric changes (in percent) and were used to enhance the classical biotype profile by adding kinetic categories. A hierarchical cluster analysis was performed by using the enhanced profiles and resulted in 14 clusters. Six major clusters grouped 94% of all isolates with a similarity of > or =95% within any given cluster, and eight clusters contained a single isolate. The six major clusters grouped not only serotypes of the same type but also phenotypic serotype variations into individual clusters. This suggests that metabolic kinetic reaction data from the biochemical tests commonly used for classic Salmonella enterica subsp. enterica biotyping can possibly be used to determine the relatedness between isolates in an easy and timely manner. PMID- 16272464 TI - In vitro activities of anidulafungin against more than 2,500 clinical isolates of Candida spp., including 315 isolates resistant to fluconazole. AB - Anidulafungin is an echinocandin antifungal agent with potent activity against Candida spp. We assessed the in vitro activity of anidulafungin against 2,235 clinical isolates of Candida spp. using the CLSI broth microdilution method. Anidulafungin was very active against Candida spp. (the MIC at which 90% of strains are inhibited [MIC(90)] was 2 microg/ml when MIC endpoint criteria of partial inhibition [MIC-2] were used). Candida albicans, C. glabrata, C. tropicalis, C. krusei, and C. kefyr were the most susceptible species of Candida (MIC(90), 0.06 to 0.12 microg/ml), and C. parapsilosis, C. lusitaniae, and C. guilliermondii were the least susceptible (MIC(90), 0.5 to 2 microg/ml). In addition, 315 fluconazole-resistant isolates were tested, and 99% were inhibited by < or =1 microg/ml of anidulafungin. These results provide further evidence for the spectrum and potency of anidulafungin activity against a large and geographically diverse collection of clinically important isolates of Candida spp. PMID- 16272465 TI - Could human papillomaviruses be spread through blood? AB - The human papillomaviruses (HPVs) are epitheliotropic viruses that require the environment of a differentiating squamous epithelium for their life cycle. HPV infection through abrasion of the skin or sexual intercourse causes benign warts and sometimes cancer. HPV DNA detected in the blood has been interpreted as having originated from metastasized cancer cells. The present study examined HPV DNA in banked, frozen peripheral blood mononuclear cells (PBMCs) from 57 U.S. human immunodeficiency virus (HIV)-infected pediatric patients collected between 1987 and 1996 and in fresh PBMCs from 19 healthy blood donors collected in 2002 to 2003. Eight patients and three blood donors were positive mostly for two subgroups of the HPV type 16 genome. The HPV genome detected in all 11 PBMC samples existed as an episomal form, albeit at a low DNA copy number. Among the eight patients, seven acquired HIV from transfusion (three associated with hemophilia) and one acquired HIV through vertical transmission; this patient also had received a transfusion before sampling. Our data suggest that PBMCs may be HPV carriers and might spread the virus through blood. PMID- 16272466 TI - Rapid detection of Mycobacterium tuberculosis in respiratory samples by transcription-reverse transcription concerted reaction with an automated system. AB - The aim of this study was to evaluate the performance of the transcription reverse transcription concerted (TRC) method for the detection of Mycobacterium tuberculosis complex (MTC) 16S rRNA in clinical respiratory samples for the diagnosis of pulmonary tuberculosis. TRC is a novel method that enables the rapid and the completely homogeneous real-time monitoring of isothermal sequence RNA amplification without any postamplification procedure. The detection limit of the TRC method for MTC was one organism per 100 mul of sputum. The specificity of the method was confirmed by the absence of positive signals for sputum containing 10(6) M. avium or M. kansasii organisms per 100 microl. A total of 201 respiratory samples from patients diagnosed with or suspected of having tuberculosis were tested. Of the 72 MTC culture-positive samples, the TRC method was positive for 52 (sensitivity, 72.2%), whereas the Roche COBAS AMPLICOR PCR was positive for 58 (sensitivity, 80.6%). Both the TRC method and the COBAS AMPLICOR PCR showed no positive identification for any of the 129 culture negative samples. The percent agreement between the two methods was 95% (191 of 201 samples). The high sensitivity and specificity together with shorter detection time (within 1 h) of the TRC method allow it to be proposed as a useful method for the rapid detection of MTC in respiratory samples. PMID- 16272467 TI - Clonal association between Streptococcus pneumoniae serotype 23A, circulating within the United States, and an internationally dispersed clone of serotype 23F. AB - Streptococcus pneumoniae is an important pathogen in the United States and is associated with significant morbidity and mortality. Since the introduction of the seven-valent conjugate vaccine, a significant decline in pneumococcal disease has been reported. However, surveillance for pneumococcal disease remains essential, as the extent of cross protection against vaccine-related serotypes is still unclear. Further, any increase in non-vaccine-related serotypes also needs monitoring. We report on a new clonal association between a vaccine-related serotype, serotype 23A, obtained as part of the Active Bacterial Core surveillance, with an established internationally dispersed Pneumococcal Molecular Epidemiology Network (PMEN) clone, clone Colombia(23F)-26. Sixty-two isolates of serotype 23A collected from sterile sites during a 2-year period (2002 and 2003) were characterized. Twenty-one (34%) isolates were penicillin nonsusceptible, although none were fully resistant. Pulsed-field gel electrophoresis and multilocus sequence typing analysis showed that 24 (39%) of the serotype 23A isolates shared either genetic identity or high genetic relatedness with PMEN clone Colombia(23F)-26. Extensive variability was noted within the sequenced region of pbp2b in two penicillin-nonsusceptible isolates as well as in PMEN clone Colombia(23F)-26, suggesting that these isolates probably acquired penicillin resistance independently. The emergence of such new serotype and genotype associations highlights the dynamic nature of the pneumococcal population, necessitating continuous monitoring in the post-vaccine era. PMID- 16272468 TI - Molecular evolution of methicillin-resistant Staphylococcus aureus in the metropolitan area of Cologne, Germany, from 1984 to 1998. AB - To investigate the molecular evolution of methicillin-resistant Staphylococcus aureus (MRSA) in a large metropolitan area in Germany, 398 nonrepetitive MRSA isolates recovered from patients from various teaching and nonteaching hospitals in Cologne between 1984 and 1998 were characterized by pulsed-field gel electrophoresis (PFGE). On this basis, 95 representative isolates were selected and further investigated by multilocus sequence typing (MLST), spa typing, and staphylococcal cassette chromosome mec (SCCmec) typing. Overall, there were 9 MLST types and 16 spa types. The most prevalent sequence types (STs) were ST239 (38% of isolates), ST247 (29%), and ST228 (18%); the most prevalent spa types were 37 (32%) and 51 (29%). ST239 comprised five major PFGE types and various unique PFGE patterns, and ST5 comprised two PFGE types. While the same PFGE pattern was not observed among strains with different STs, spa type 37 was observed among strains representing two different STs (ST239 and ST241), and these belonged to the same clonal complex as single-locus variants. ST239 was the earliest predominant ST, with the highest prevalence from 1984 to 1988 (96%), followed by ST247 from 1989 to 1993 (83%) and ST228 from 1994 to 1998 (40%). Spa type 37 was the most prevalent from 1984 to 1988 (96%), spa type 51 was the most prevalent from 1989 to 1993 (83%), and spa types 1 and 458 were the most prevalent from 1994 to 1998 (26% and 14%, respectively). The prevalence of SCCmec type III decreased from 96% from 1984 to 1988 to 8% from 1989 to 1993, the prevalence of SCCmec type I increased from 4% from 1984 to 1988 to 97% from 1989 to 1993 and decreased to 62% from 1994 to 1998. While the genetic diversity of MRSA increased from 1984 to 1998, one prevalent ST usually accounted for most of the isolates in a given time period. PMID- 16272469 TI - Development of one-step, real-time, quantitative reverse transcriptase PCR assays for absolute quantitation of human coronaviruses OC43 and 229E. AB - The clinical significance of human coronaviruses in more severe respiratory illnesses has recently been shown to be higher than was previously assumed. Rapid and reliable diagnosis of human coronavirus infections therefore becomes indispensable in a routine clinical setting. In this study, we present a very sensitive and specific TaqMan-based, real-time quantitative reverse transcriptase PCR (qRT-PCR) for the rapid detection and quantitation of human coronaviruses (HCoVs) OC43 and 229E. Absolute viral load measurement in clinical samples was achieved through the construction of in-house HCoV OC43 and 229E cRNA standards for the generation of a standard curve. The HCoV OC43 assay allows quantitation over a range from 20 to 2 x 10(8) RNA copies per reaction mixture (5 microl RNA extract). When this is extrapolated to clinical samples, this corresponds to a detection range of 10(3) to 10(10) viral genome equivalents per ml. By using the HCoV 229E qRT-PCR assay, viral RNA copies ranging from 200 to 2 x 10(9) per reaction mixture can be detected, which corresponds to 10(4) to 10(11) viral genome equivalents per ml sample. A total of 100 respiratory samples screened for the presence of HCoVs OC43 and 229E by using conventional RT-PCR were assessed in parallel by the qRT-PCR assays. By use of the real-time qRT-PCR techniques, the detection rate of HCoVs OC43 and 229E increased from 2.0% to 3.1% and from 0.3% to 2.5%, respectively. The real-time qRT-PCR assays described here allow the rapid, specific, and sensitive laboratory detection and quantitation of human coronaviruses OC43 and 229E. PMID- 16272470 TI - Antigenic divergence of Bordetella pertussis isolates in Taiwan. AB - In recent studies, antigenic divergence has been observed in Bordetella pertussis circulating isolates. We collected 80 Bordetella pertussis isolates in Taiwan from 1998 to 2004 and analyzed them using a combination of pulsed-field gel electrophoresis (PFGE) and sequencing of the ptxS1 and prn genes. The incidence of pertussis increases every 3 years, and most of the isolates prevalent since 1998 have expressed nonvaccine ptxS1A and prn2 alleles. Through PFGE analysis, all isolates could be classified into four major groups, and the incidence of these groups exhibited a correlation with the prn allele expressed by the isolates. We found that PFGE is more discriminative than gene sequencing, since it could divide the isolates expressing the prn2 allele into two groups: one group circulating from 1998 to 2001 and another group circulating from 2001 to 2004. The transition between the two groups in 2000 coincided with an outbreak of 326 cases. This research indicates that the antigenic divergence of B. pertussis circulating isolates has evolved over time in Taiwan. Such information will have implications for vaccine policy in Taiwan. PMID- 16272471 TI - Characterization of a serodiagnostic complement fixation antigen of Coccidioides posadasii expressed in the nonpathogenic Fungus Uncinocarpus reesii. AB - Coccidioides spp. (immitis and posadasii) are the causative agents of human coccidioidomycosis. In this study, we developed a novel system to overexpress coccidioidal proteins in a nonpathogenic fungus, Uncinocarpus reesii, which is closely related to Coccidioides. A promoter derived from the heat shock protein gene (HSP60) of Coccidioides posadasii was used to control the transcription of the inserted gene in the constructed coccidioidal protein expression vector (pCE). The chitinase gene (CTS1) of C. posadasii, which encodes the complement fixation antigen, was expressed using this system. The recombinant Cts1 protein (rCts1(Ur)) was induced in pCE-CTS1-transformed U. reesii by elevating the cultivation temperature. The isolated rCts1(Ur) showed chitinolytic activity that was identical to that of the native protein and had serodiagnostic efficacy comparable to those of the commercially available antigens in immunodiffusion complement fixation tests. Using the purified rCts1(Ur), 74 out of the 77 coccidioidomycosis patients examined (96.1%) were positively identified by enzyme linked immunosorbent assay. The rCts1(Ur) protein showed higher chitinolytic activity and slightly greater seroreactivity than the bacterially expressed recombinant Cts1. These data suggest that this novel expression system is a useful tool to produce coccidioidal antigens for use as diagnostic antigens. PMID- 16272472 TI - Cultivable oral microbiota of domestic dogs. AB - Bacteria were isolated from the dental plaques of nine dogs and a sample of pooled saliva from five other dogs and were then identified by comparative 16S rRNA gene sequencing. Among 339 isolates, 84 different phylotypes belonging to 37 genera were identified. Approximately half of the phylotypes were identified to the species level, and 28% of these were considered members of the indigenous oral microbiota of humans. The 16S rRNA gene sequences of the remaining 44 phylotypes were not represented in GenBank, and most of these phylotypes were tentatively identified as candidate new species. The genera most frequently isolated from saliva were Actinomyces (26%), Streptococcus (18%), and Granulicatella (17%). The genera most frequently isolated from plaque were Porphyromonas (20%), Actinomyces (12%), and Neisseria (10%). A comparison of the DNA sequences from this study with sequences of the human microbiota available in GenBank showed that, on average, canine and human microbiotas differed by almost 7% in the 16S rRNA gene. In conclusion, this study has shown that the cultivable oral microbiotas of dogs and humans show significant differences. PMID- 16272473 TI - Detection of mutations associated with isoniazid resistance in Mycobacterium tuberculosis isolates from China. AB - Nine structural genes (furA, katG, inhA, kasA, Rv0340, iniB, iniA, iniC, and efpA) and two regulatory regions (the oxyR-ahpC intergenic region and the promoter of mabA-inhA) in 87 isoniazid (INH)-monoresistant and 50 INH-susceptible Mycobacterium tuberculosis isolates collected from five provinces of China were analyzed by sequencing. Eighty-two (94.3%) INH-resistant isolates had mutations in the katG gene, with the katG Ser315Thr mutation predominant (55.2%). No mutation at codon 463 of katG was detected among the 50 INH-susceptible isolates with different IS6110 fingerprints. In addition, there were 35 (40.2%) INH resistant isolates that had a mutation at codon 463 of katG. Of the INH-resistant strains, 20 (23.0%) isolates harbored double mutations at two separate loci of katG. Mutations in the inhA promoter region occurred in 13 (14.9%) isolates; 4.6% of the isolates had inhA structural gene mutations, and 11.5% harbored mutations in the oxyR-ahpC intergenic region. Drug resistance-associated mutations were detected in the iniBAC region and efpA. PMID- 16272474 TI - Assessment of band-based similarity coefficients for automatic type and subtype classification of microbial isolates analyzed by pulsed-field gel electrophoresis. AB - Pulsed-field gel electrophoresis (PFGE) has been the typing method of choice for strain identification in epidemiological studies of several bacterial species of medical importance. The usual procedure for the comparison of strains and assignment of strain type and subtype relies on visual assessment of band difference number, followed by an incremental assignment to the group hosting the most similar type previously seen. Band-based similarity coefficients, such as the Dice or the Jaccard coefficient, are then used for dendrogram construction, which provides a quantitative assessment of strain similarity. PFGE type assignment is based on the definition of a threshold linkage value, below which strains are assigned to the same group. This is typically performed empirically by inspecting the hierarchical cluster analysis dendrogram containing the strains of interest. This approach has the problem that the threshold value selected is dependent on the linkage method used for dendrogram construction. Furthermore, the use of a linkage method skews the original similarity values between strains. In this paper we assess the goodness of classification of several band-based similarity coefficients by comparing it with the band difference number for PFGE type and subtype classification using receiver operating characteristic curves. The procedure described was applied to a collection of PFGE results for 1,798 isolates of Streptococcus pneumoniae, which documented 96 types and 396 subtypes. The band-based similarity coefficients were found to perform equally well for type classification, but with different proportions of false-positive and false negative classifications in their minimal false discovery rate when they were used for subtype classification. PMID- 16272475 TI - Comparison of real-time PCR protocols for differential laboratory diagnosis of amebiasis. AB - Specific identification of Entamoeba spp. in clinical specimens is an important confirmatory diagnostic step in the management of patients who may be infected with Entamoeba histolytica, the species that causes clinical amebiasis. Distinct real-time PCR protocols have recently been published for identification of E. histolytica and differentiation from the morphologically identical nonpathogenic Entamoeba dispar. In this study, we compared three E. histolytica real-time PCR techniques published by December 2004. The limits of detection and efficiency of each real-time PCR assay were determined using DNA extracted from stool samples spiked with serially diluted cultured E. histolytica trophozoites. The ability of each assay to correctly distinguish E. histolytica from E. dispar was evaluated with DNA extracted from patients' stools and liver aspirates submitted for confirmatory diagnosis. Real-time PCR allowed quantitative analysis of the spiked stool samples, but major differences in detection limits and assay performance were observed among the evaluated tests. These results illustrate the usefulness of comparative evaluations of diagnostic assays. PMID- 16272476 TI - Use of full-length recombinant calflagin and its c fragment for improvement of diagnosis of Trypanosoma cruzi infection. AB - Serological diagnosis of Trypanosoma cruzi infection is hampered by issues related to test specificity due to the cross-reactivity of most antigens with proteins of related parasites such as Leishmania spp. The recombinant calflagins are considered relevant antigens for the diagnosis of infection by Trypanosoma cruzi. In the present work, we describe two genes coding for putative calflagins in Leishmania major with the N-terminal moieties presenting high similarity with T. cruzi genes. This fact raised questions about their role in some cross recognition of this antigen by sera from Leishmania spp.-infected individuals. The complete T. cruzi calflagin and two fragments of the protein, consisting of 146 amino acids of the N-terminal and 65 amino acids of the C-terminal regions, were expressed and evaluated against a panel of sera, which included well characterized samples from T. cruzi, and Leishmania-infected patients. We were able to show that sera from Leishmania (Viannia) braziliensis-infected individuals recognized the recombinant full-length calflagin. Both the N-terminal and the complete protein presented the same high sensitivity (98.5% of sera from T. cruzi-infected patients was detected) but different specificities (94% and 98%, respectively, when evaluated against sera from people not infected by T. cruzi, including 15 sera from people infected with L. braziliensis). The C terminal fragment presented low sensitivity (70%) but 100% specificity. We propose the use of these antigens in two sequential assays to optimize the serological diagnosis of T. cruzi infection in humans in geographic areas where Leishmania spp. infection is coendemic. PMID- 16272477 TI - A novel bacterial mucinase, glycosulfatase, is associated with bacterial vaginosis. AB - The modifications to the vaginal habitat accompanying a change to vaginal flora in bacterial vaginosis (BV) are poorly understood. In this study enzymes involved in mucin degradation were measured, including a novel glycosulfatase assay. Women attending an emergency walk-in sexually transmitted disease clinic were studied. One high vaginal swab (HVS) was used to prepare a gram-stained smear to determine BV status, using Ison and Hay's criteria, and a separate swab was used for the purposes of the assays. The median glycosulfatase activity was 8.5 (range, -1.2 to 31.9) nmol h(-1) 1.5 ml(-1) of HVS suspension in patients with BV compared to 0.5 (range, -0.7 to 9.4) nmol h(-1) 1.5 ml(-1) of HVS suspension in patients without BV (P = <0.001). The median glycoprotein sialidase activity was 29.2 (range, -17 to 190) nmol h(-1) 1.5 ml(-1) of HVS suspension in patients with BV compared to -1.1 (range, -41 to 48) nmol h(-1) 1.5 ml(-1) of HVS suspension in patients without BV (P < 0.001). A rapid spot test for sialidase was positive in 22/24 patients with BV (sensitivity, 91.7%; 95% confidence interval [CI], 73 to 99%) and negative in 32/35 patients without BV (specificity, 91.4%; 95% CI, 76.9 to 98.2%) (P < 0.001). Glycosulfatase activity significantly correlated with both glycoprotein sialidase activity and the sialidase spot test (P = 0.006 and P < 0.001, respectively). The results are consistent with the hypothesis that the consortium of bacteria present in BV requires the ability to break down mucins in order to colonize the vagina and replace the normal lactobacilli. PMID- 16272478 TI - Rapid identification of human adenovirus types 3 and 7 from respiratory specimens via multiplex type-specific PCR. AB - The rapid diagnosis of human adenovirus (Ad) infection is crucial for the timely recognition of epidemics. Moreover, identification of the serotypes known to cause serious disease can be helpful in therapeutic intervention. A multiplex PCR assay was developed for the rapid detection of adenovirus type 3 (Ad3) and Ad7 directly from clinical specimens. For this assay, three primer pairs (primers were based on the conserved and hypervariable regions of the hexon) were designed in order to simultaneously amplify all adenoviral serotypes and discriminate between Ad3 and Ad7. In our preliminary analysis, this multiplex PCR assay generated amplicons of the consensus primers from all 106 adenoviral isolates of diverse serotypes and proved able to correctly identify Ad3 and Ad7. This assay was subsequently applied to the detection of Ad3 and Ad7 in respiratory specimens. Among the 127 nasal aspirates from which an adenovirus was grown, the sensitivity with which any serotype could be detected was 91% (115/127). Two of the 53 nasal aspirates which did not grow Ads yielded adenovirus-specific bands, which were confirmed by sequencing analysis. Among the 115 specimens which produced common adenoviral bands, the sensitivity with which Ad3 could be detected was 93% (26/28), and the sensitivity with which Ad7 could be detected was 100% (35/35). Five out of the 115 specimens were proved to harbor more than one type of Ad via sequencing analysis of the amplicons, suggesting mixed infection with at least two different serotypes. In conclusion, this multiplex PCR system can be utilized in the rapid identification of Ad3 and Ad7 directly from clinical specimens. Furthermore, this method constitutes a diagnostic strategy for the detection of coinfection by different Ad serotypes. PMID- 16272479 TI - Evaluation of enzyme-linked immunosorbent assays, an immunofluorescent-antibody test, and two rapid tests (immunochromatographic-dipstick and gel tests) for serological diagnosis of symptomatic and asymptomatic Leishmania infections in dogs. AB - Enzyme-linked immunosorbent assays (ELISAs) based on soluble antigens derived from promastigote or amastigote-like stages of Leishmania infantum and on the recombinant rK39 antigen, each in combination with different conjugates [anti immunoglobulin G1 [IgG1], anti-IgG2, anti-IgG(gamma), and anti-IgG heavy plus light chains], were compared to an immunofluorescent-antibody test (IFAT) and two commercially available rapid test systems (DiaMed-Vet-IT Leish and ID-PaGIA canine leishmaniasis antibody test) for the detection of specific anti-Leishmania antibodies in symptomatic and asymptomatic dogs with proven L. infantum infections. ELISAs based on soluble promastigote and amastigote antigens had very high sensitivities in symptomatic (n = 30; 100%) and asymptomatic dogs (n = 17; 94.1 to 100%), except when combined with the anti-IgG1 conjugate (41.2 to 82.4%). Specificities were high for all combinations (n = 50; 96 to 100%). The rK39 ELISA detected fewer asymptomatic cases (sensitivities, 52.9 to 64.7%) but was highly specific (96 to 100%). The IFAT was 90% sensitive in symptomatic dogs but was significantly less sensitive in asymptomatic cases (29.4%). However, it had an excellent specificity (100%). Test performances of the rapid tests based on the rK39 antigen were comparable to the ELISAs based on the same antigen. ELISAs based on soluble promastigote or amastigote antigens seem to be most suited for the serological diagnosis of canine Leishmania infections in both symptomatic and asymptomatic dogs. IFAT and the rK39 ELISA lack sensitivity in asymptomatic cases but are highly specific. Rapid tests like the rK39 dipstick test or the ID-PaGIA are helpful for confirming clinically suspected cases because of their high specificities in symptomatic animals. PMID- 16272480 TI - Epidemiological survey of human metapneumovirus infection in a large pediatric tertiary care center. AB - The human metapneumovirus (hMPV) was recently identified and linked to acute respiratory tract infections (ARTI). To assess the clinical importance of this virus in infants and children, we developed a rapid and efficient reverse transcription-PCR-based screening method for a large volume of samples and tested retrospectively a collection of 1,132 respiratory specimens submitted over a full year period to the virology laboratory of a large tertiary care pediatric center in Montreal, Canada. A total of 41 samples from 37 patients were positive by this method. During the winter months of 2001, up to 8% of specimens submitted for respiratory virus testing were hMPV positive. Sequencing data of the hMPV M gene revealed that two genogroups of the virus, each of which can be divided into two subgroups, cocirculated during this time period. A case-controlled study was conducted to compare the symptoms associated with hMPV infection with those involving other etiologic agents causing ARTI. Symptoms most frequently observed in hMPV-positive patients were cough, wheezing, and dyspnea, although the symptomatology could differ substantially from patient to patient. No distinct symptom profile could be associated with hMPV. Three nosocomial cases of hMPV infection were identified. Together, our data suggest that hMPV is a significant cause of symptomatic respiratory tract infections in infants and children. The incidence of the disease and the morbidity associated with the infection justify adding hMPV to the list of common respiratory viruses routinely screened for by clinical laboratories. PMID- 16272481 TI - Analysis of the effect of DNA purification on detection of human papillomavirus in oral rinse samples by PCR. AB - Human papillomavirus (HPV) has recently been associated with oral cancers. To prepare for a study of the natural history of oral HPV infection, the effect of the DNA purification method on HPV genomic DNA detection in Scope mouthwash oral rinse samples and the reproducibility of HPV detection in rinse samples collected 7 days apart were investigated. The study was conducted with a population at high risk for oral HPV infection: human immunodeficiency virus-infected men with CD4 cell counts <200. Five DNA purification methods were compared among equal aliquots of oral rinse samples collected from a subset of individuals. The purification methods included (i) proteinase K digestion (PKD) and heat inactivation; (ii) PKD and ethanol precipitation (EP); (iii) PKD, phenol chloroform extraction, and EP; (iv) use of the Puregene DNA purification kit; and (v) use of the QIAamp DNA Blood Midi kit. HPV was detected by PCR amplification with PGMY09 and PGMY11 L1 primer pools and by use of a Roche linear array. Puregene-purified samples had higher human DNA yields and purities, and Puregene purification detected the greatest number of HPV-positive subjects and total HPV infections in comparison to the numbers detected by all other methods. The total number of HPV infections and HPV prevalence estimates were also higher for Puregene-processed oral rinse samples when a fixed volume (10 mul) rather than a fixed cell number ( approximately 50,000 cells) was used for PCR amplification. A good concordance was observed for oral HPV infection status (agreement, 80%; kappa value, = 0.60) and type-specific infection (agreement, 98%; kappa value, 0.57) in matched oral rinse samples. The method of DNA purification significantly affects the detection of HPV genomic DNA from oral rinse samples and may result in exposure misclassification that could contribute to the inconsistent associations reported in the literature. PMID- 16272482 TI - Multicenter evaluation of BBL CHROMagar MRSA medium for direct detection of methicillin-resistant Staphylococcus aureus from surveillance cultures of the anterior nares. AB - Active surveillance for methicillin-resistant Staphylococcus aureus (MRSA) is among the strategies recommended by the Society for Healthcare Epidemiology of America for control of nosocomial MRSA infections. Infection control and laboratory personnel desire rapid, sensitive, and inexpensive methods to enhance surveillance activities. A multicenter study was performed to evaluate a new selective and differential chromogenic medium, BBL CHROMagar MRSA (C-MRSA) medium (BD Diagnostics, Sparks, MD), which enables recovery and concomitant identification of MRSA strains directly from nasal swab specimens taken from the anterior nares. Specimens were inoculated to C-MRSA and Trypticase soy agar with 5% sheep blood agar (TSA II, BD Diagnostics). Mauve colonies on C-MRSA at 24 h and 48 h and suspicious colonies on TSA II were confirmed as Staphylococcus aureus by Gram stain morphology and a coagulase test. In addition, the results of C-MRSA were compared to results of susceptibility testing (five different methods) of S. aureus strains isolated on TSA II. A total of 2,015 specimens were inoculated to C-MRSA and TSA II. Three hundred fifty-four S. aureus isolates were recovered; 208 (59%) were oxacillin (methicillin) susceptible and 146 (41%) were oxacillin resistant (MRSA). On C-MRSA, 139/146 or 95.2% of MRSA isolates were recovered, whereas recovery on TSA II was 86.9% (127/146) (P = 0.0027). The overall specificity of C-MRSA was 99.7%. When C-MRSA was compared to each susceptibility testing method, the sensitivity and specificity, respectively, were as follows: oxacillin MIC by broth microdilution, 94.4% and 96.7%; oxacillin screen agar, 94.3% and 96.7%; PBP2' latex agglutination, 93.7% and 98.5%; cefoxitin disk diffusion, 95.0% and 98.1%; and mecA PCR, 95.1% and 98.1%. In this study, C-MRSA was superior to TSA II for recovery of MRSA from surveillance specimens obtained from the anterior nares and was comparable to conventional, rapid, and molecular susceptibility methods for the identification of MRSA isolates. PMID- 16272483 TI - Automated extraction of viral-pathogen RNA and DNA for high-throughput quantitative real-time PCR. AB - The performance of the m1000 system (Abbott Laboratories, Illinois) as a front end extraction system for high-throughput "in-house" quantitative real-time PCR assays was analyzed and compared to that of manual extraction of plasma and serum samples (hepatitis C virus [HCV] and hepatitis B virus [HBV]) and EDTA-blood samples (cytomegalovirus [CMV] and Epstein-Barr virus [EBV]). Linearity of extraction was tested on dilution series of HCV and HBV reference materials. The correlation coefficient for standard curves based on repeated extraction runs was 0.97 +/- 0.06 for HCV and 0.97 +/- 0.03 for HBV, indicating a linear extraction from 100 to 1.0 x 10(5) HCV IU/ml and from 100 to 1.0 x 10(6) HBV IU/ml. Intra- and interrun variability was below 0.23 log(10) IU/ml for 2.98 to 5.28 log(10) HCV IU/ml and 2.70 to 5.20 log(10) HBV IU/ml. Correlation between automated and manual extraction was very good. For HCV, the correlation coefficient was 0.91 and the mean difference in viral load was 0.13 log(10) HCV IU/ml. For HBV, the correlation coefficient was 0.98 and the mean difference in viral load 0.61 log(10) HBV IU/ml. For CMV and EBV, the correlation coefficient was 0.98 and the mean difference in viral load 0.33 log(10) copies/ml. Accuracy was confirmed with a reference panel (QCMD, Glasgow, Scotland) for all four assays. No cross contamination was observed when extracting strongly positive polyomavirus samples (8.10 log(10) copies/ml) interspersed with polyomavirus-negative samples. Automated extraction via the m1000 system offers a high reliability of extraction and resulted in a strong reduction of the required extraction hands-on time for high-throughput PCR compared to manual extraction protocols. PMID- 16272485 TI - Use of rapid genomic deletion typing to monitor a tuberculosis outbreak within an urban homeless population. AB - Beginning in mid-2002, a large tuberculosis outbreak occurred among homeless persons in King County, Washington. In order to further monitor the outbreak following its peak in 2003, Mycobacterium tuberculosis isolates from all new King County tuberculosis (TB) patients in 2004 and the first half of 2005 (n = 220) were genotyped by using a rapid comparative genomics-based (genomic deletion typing) approach, with confirmation by mycobacterial interspersed repetitive units and repetitive-sequence-based PCR (rep-PCR). Results were compared to retrospective genotypic data from 1995 to 2003. The outbreak strain SBRI9, which was not seen among King County homeless persons prior to 2002, accounted for 16 out of 30 TB cases (53%) within this population in 2002. This trend continued with 27 out of 35 cases (77%) caused by the outbreak strain in 2003, 11 out of 13 cases (85%) caused by the outbreak strain in 2004, and 4 out of 10 cases (40%) caused by the outbreak strain in the first 5 months of 2005. Thus, the outbreak strain remained well established within this homeless population throughout the study period. At least four SBRI9 cases were in people who had previously been infected by other strains. The novel PCR-based strain-typing approach used in this investigation proved to be cost-effective and very rapid. In most cases, it was possible to analyze DNA extracted directly from primary isolation (Mycobacterium growth indicator tube) cultures submitted by clinical laboratories, a feature that markedly reduced the delay between diagnosis and strain typing results. This rapid turnaround facilitated public health efforts to prevent new outbreaks involving this strain. PMID- 16272484 TI - Relationship between MIC and minimum sterol 14{alpha}-demethylation-inhibitory concentration as a factor in evaluating activities of azoles against various fungal species. AB - The minimum growth-inhibitory concentrations (MICs) of azole antifungals were compared to their minimum sterol 14alpha-demethylation-inhibitory concentrations (MDICs) for clinical fungal isolates. The ascomycetous Candida yeasts tested were clearly divided into two groups: group I, consisting of C. albicans, C. tropicalis, and C. lusitaniae, had MICs that were much higher than the MDICs, whereas group II, comprising C. glabrata, C. parapsilosis, C. guilliermondii, and C. krusei, had MICs that were approximately equal to the MDICs. In the ascomycetous fungi Aspergillus fumigatus and Sporothrix schenckii, the MICs were indistinguishable from the MDICs. In the basidiomycetous fungi Cryptococcus (Filobasidiella) neoformans, C. curvatus, and Trichosporon asahii, the MICs and the MDICs were practically identical. These results support the notion that there are two distinct classes of fungi differing in their degree of tolerance to sterol 14alpha-demethylation deficiency. These findings have significant implications for both fungal physiology and antifungal chemotherapy. PMID- 16272486 TI - New criteria for immunofluorescence assay for Q fever diagnosis in Japan. AB - A study was made to evaluate the cutoff value of indirect immunofluorescent antibody (IFA) test for Q fever diagnosis in Japan. We used 346 sera, including 16 from confirmed Q fever cases, 304 from Japanese pneumonia patients, and 26 from negative cases. Thirteen sera from the confirmed Q fever cases with an immunoglobulin M (IgM) titer of > or =1:128 and/or IgG titer of > or =1:256 by the IFA test were positive by both enzyme-linked immunosorbent assay (ELISA) and Western blotting assay (WBA), whereas 298 sera from pneumonia patients and 26 negative sera with an IgM titer of < or =1:16 and an IgG titer of < or =1:32 by the IFA test were negative by both ELISA and WBA. In the proposed "equivocal area," with an IgM titer of > or =1:32 and < or =1:64 and/or an IgG titer of > or =1:64 and < or =1:128, we found 9 sera, 3 from confirmed Q fever cases and 6 from Japanese pneumonia patients, by the IFA test. Three sera from the confirmed Q fever cases and one of the sera from pneumonia patients were IgM and/or IgG positive by both ELISA and WBA. These results suggest that a single cutoff value for the IFA test may cause false-positive and false-negative results. In conclusion, this study showed that an "equivocal area" should be used for the IFA test rather than a single cutoff value and that sera in the equivocal area should be tested by additional serological assays for confirmation. PMID- 16272487 TI - Quantitative nucleic acid sequence-based assay as a new molecular tool for detection and quantification of Leishmania parasites in skin biopsy samples. AB - Currently available methods for the diagnosis of cutaneous leishmaniasis (CL) have low sensitivities or are unable to quantify the number of viable parasites. This constitutes a major obstacle for the diagnosis of the disease and for the study of the effectiveness of treatment schedules and urges the development of improved detection methods. In this study, quantitative nucleic acid sequence based amplification (QT-NASBA) technology was used to detect and quantify Leishmania parasites in skin biopsy samples from CL patients. The assay is based on the detection of a small subunit rRNA (18S rRNA), which may allow for the detection of viable parasites. The QT-NASBA assay was evaluated using in vitro cultured promastigotes and amastigotes and 2-mm skin biopsy samples from Old and New World CL patients. The study demonstrated that the lower detection limit of the QT-NASBA was two parasites per biopsy sample. Parasites could be quantified in a range of 2 to 11,300,000 parasites per biopsy sample. The QT-NASBA could detect levels of parasites 100-fold lower than those detected by conventional PCR. Test evaluation revealed that the QT-NASBA had a sensitivity of 97.5% and a specificity of 100% in the present study. The QT-NASBA is a highly sensitive and specific method that allows quantification of both Old and New World Leishmania parasites in skin biopsy samples and may provide an important tool for diagnosis as well as for monitoring the therapy of CL patients. PMID- 16272488 TI - Analytical verification of a PCR assay for identification of Bordetella avium. AB - Bordetella avium is the etiologic agent of turkey coryza or bordetellosis, a respiratory disease responsible for substantial economic losses to the turkey industry. At present, identification of this bacterium relies on isolation and biochemical testing. Although a PCR for the detection of B. avium was proposed a number of years ago, lack of analytical verification precludes its use as a diagnostic tool. Furthermore, a number of details pertaining to the reaction conditions used are missing or unclear. In the present study we have identified an optimal set of PCR conditions for use with the previously described primer pair and determined the limit of detection under these conditions to be approximately 20 pg. Assay sensitivity is 100%, based on an analysis of 72 B. avium isolates from diverse geographic locations and covering a time span of at least 25 years. Evaluation of a separate group of 87 bacterial isolates from poultry, comprising both gram-positive and gram-negative commensals and pathogens representing 11 genera, demonstrated an assay specificity of 98.8%. Reproducibility is 100% using either purified genomic DNA or boiled cell lysates less than 3 days old. Sequence analysis of the B. avium PCR amplicons identified only three occasional sequence polymorphisms. These data indicate the B. avium PCR assay can provide clinically significant results. PMID- 16272489 TI - TaqMan real-time reverse transcription-PCR and JDVp26 antigen capture enzyme linked immunosorbent assay to quantify Jembrana disease virus load during the acute phase of in vivo infection. AB - Jembrana disease virus (JDV) is an acutely pathogenic lentivirus that affects Bali cattle in Indonesia. The inability to propagate the virus in vitro has made it difficult to quantitate JDV and determine the kinetics of virus replication during the acute phase of the disease process. We report for the first time two techniques that enable quantification of the virus and the use of these techniques to quantify the virus load during the acute phase of the disease process. A one-step JDV gag [corrected] TaqMan real-time reverse transcription PCR (RT-PCR) assay was developed for the detection and quantification of JDV RNA in plasma. The limit of detection was 9.8 x 10(2) JDV viral RNA copies over 35 cycles, equivalent to 4.2 x 10(4) JDV genome copies/ml, and a peak virus load of 1.6 x 10(12) during the acute febrile period. An antigen capture enzyme-linked immunosorbent assay (ELISA) was also developed to quantify the levels of JDV capsid (JDVp26) over a linear range of 10 to 200 ng/ml. Viral RNA and JDVp26 levels were correlated in 48 plasma samples obtained from experimentally infected cattle. A significant positive correlation (R = 0.860 and r(2) = 0.740) was observed between the two techniques within the range of their detection limits. The relatively insensitive capture ELISA provides an economical and feasible method for monitoring of virus in the absence of more sensitive techniques. PMID- 16272490 TI - Development of a general-primer-PCR-reverse-line-blotting system for detection of beta and gamma cutaneous human papillomaviruses. AB - The beta and gamma genera of papillomaviruses consist of epidermodysplasia verruciformis-related human papillomaviruses (HPVs) and phylogenetically related cutaneous HPVs. Here, we have developed a consensus primer PCR assay and reverse line blot typing system coupled thereto (referred to as beta and gamma cutaneous HPV PCR [BGC-PCR]) for detection and typing of 24 beta and gamma HPVs (HPV types 4, 5, 8, 9, 12, 14, 15, 17, 19, 20, 21, 22, 23, 24, 25, 36, 37, 38, 47, 48, 49, 50, 60, and 65). Because the HPV-specific PCR products are only 72 bp in size, the system is suitable for formalin-fixed, paraffin-embedded specimens and other samples in which the DNA is of suboptimal quality. This system was able to detect and type as little as 100 ag to 1 fg HPV DNA per reaction (depending on the HPV type) in a background of 100 ng human DNA without any cross-reactivity between the tested types. Beta and gamma HPVs were detected in DNA extracted from plucked eyebrow hairs of 31 of 34 renal transplant recipients. In addition, formalin fixed, paraffin-embedded specimens from nonmelanoma skin tumors of renal transplant recipients (n = 25) and immunocompetent individuals (n = 15) scored BGC-PCR positive in 21 and 6 cases, respectively, with HPV type 5 (HPV5) and HPV8 being the predominant types. The data indicate that this method can be a valuable, user-friendly tool for the detection and typing of cutaneous HPV in clinical specimens and may have implications for future monitoring of vaccines or alternative treatment modalities for diseases caused by these cutaneous HPVs. PMID- 16272491 TI - Resilience of the dominant human fecal microbiota upon short-course antibiotic challenge. AB - Recent studies have shown that the human fecal microbiota is composed of a consortium of species specific to the host and resistant to modifications over time. Antibiotics are known to affect the intestinal microflora, and ensuing changes may result in antibiotic-associated diarrhea. It is therefore important to characterize the nature and amplitude of these modifications and the ability of this ecosystem to return to its original profile-i.e., its resilience. Six healthy volunteers received oral amoxicillin (1.5 g/day) for 5 days. Fecal samples were collected at day 0 (D0) before antibiotic treatment and at set intervals until 60 days thereafter. Fecal DNA was isolated, and V6-to-V8 regions of the 16S rRNA genes were amplified by PCR with general primers and analyzed by temporal temperature gradient gel electrophoresis. Dominant species profiles were compared on the basis of similarity (Pearson correlation coefficient). Dominant species profiles at D0 were used as a reference. The fecal microbiota showed a major shift in dominant species upon antibiotic treatment, starting 24 h after treatment initiation and reaching an average similarity of only 74% after 4 days. Within 30 days following antibiotic treatment, the fecal microbiota tended to reach an average similarity of 88% to the D0 value; within 60 days, the average similarity to the D0 value was 89%. However, in one subject, important modifications persisted for at least 2 months, with similarity to the D0 value remaining below 70%. We demonstrated the resilience of the dominant human fecal microbiota upon short-course antibiotic challenge. Yet the persistence of long term alterations in some subjects may explain susceptibilities to antibiotic associated diarrhea. Furthermore, these findings suggest that strategies reinforcing the ability of the fecal microbiota to resist modifications would be of clinical relevance. PMID- 16272492 TI - Multilocus sequence typing for differentiation of strains of Candida tropicalis. AB - A system is described for typing isolates of the pathogenic fungus Candida tropicalis, based on sequence polymorphisms in fragments of six genes: ICL1, MDR1, SAPT2, SAPT4, XYR1, and ZWF1a. The system differentiated 87 diploid sequence types (DSTs) among a total of 106 isolates tested or 80 DSTs among 88 isolates from unique sources. Replicate isolates from the same source clustered together with high statistical similarity, with the exception of one isolate. However, a clade of very closely related isolates included replicate isolates from three different patients, as well as single isolates from eight other patients. This clade, provisionally designated clade 1, was one of three clusters of isolates with high statistical similarity. Five of six isolates in one cluster that may acquire clade status were resistant to flucytosine. This study adds C. tropicalis to Candida albicans and Candida glabrata as Candida species for which a multilocus sequence typing (MLST) system has been set up. The C. tropicalis MLST database can be accessed at http://pubmlst.org/ctropicalis/. PMID- 16272493 TI - Population structure and properties of Candida albicans, as determined by multilocus sequence typing. AB - We submitted a panel of 416 isolates of Candida albicans from separate sources to multilocus sequence typing (MLST). The data generated determined a population structure in which four major clades of closely related isolates were delineated, together with eight minor clades comprising five or more isolates. By Fisher's exact test, a statistically significant association was found between particular clades and the anatomical source, geographical source, ABC genotype, decade of isolation, and homozygosity versus heterozygosity at the mating type-like locus (MTL) of the isolates in the clade. However, these associations may have been influenced by confounding variables, since in a univariate analysis of variance, only the clade associations with ABC type and anatomical source emerged as statistically significant, providing the first indication of possible differences between C. albicans strain type clades and their propensity to infect or colonize different anatomical locations. There were no significant differences between clades with respect to distributions of isolates resistant to fluconazole, itraconazole, or flucytosine. However, the majority of flucytosine-resistant isolates belonged to clade 1, and these isolates, but not flucytosine-resistant isolates in other clades, bore a unique mutation in the FUR1 gene that probably accounts for their resistance. A significantly higher proportion of isolates resistant to fluconazole, itraconazole, and flucytosine were homozygous at the MTL, suggesting that antifungal pressure may trigger a common mechanism that leads both to resistance and to MTL homozygosity. The utility of MLST for determining clade assignments of clinical isolates will form the basis for strain selection for future research into C. albicans virulence. PMID- 16272494 TI - Persistent detection of varicella-zoster virus DNA in a previously healthy child after severe chickenpox. AB - In immunocompetent children with primary varicella-zoster virus (VZV) infection, peak viral loads are detected in peripheral blood near the onset of the vesicular rash. VZV DNA concentrations normally diminish and become undetectable within 3 weeks after the appearance of the exanthem. Here, we present a previously healthy, human immunodeficiency virus-negative, 4-year-old boy admitted with severe varicella. High viral loads (>340,000 copies/ml) were found in his blood, and the viral loads remained high for at least 1.5 years. Clinical recovery preceded complete clearance of the virus. General and VZV-specific immune reactivity were intact. NK cells and CD8(+) T cells were activated during acute infection, and VZV-specific CD4(+) T cells were detected at high frequencies. VZV DNA was initially detected in B cells, NK cells, and both CD4(+) and CD8(+) T cells. In contrast, during the persistent phase of VZV DNA detection, the viral DNA was primarily located in CD8(+) T cells. For the first time, we describe the persistent detection of VZV DNA in a previously healthy child. PMID- 16272495 TI - Fast ligation-mediated PCR, a fast and reliable method for IS6110-based typing of Mycobacterium tuberculosis complex. AB - IS6110 restriction fragment length polymorphism (RFLP) analysis is the most widely applied method for strain differentiation of Mycobacterium tuberculosis complex. We have previously described mixed-linker PCR, an IS6110-based PCR method that favorably compared with other typing methods for M. tuberculosis complex according to reproducibility and ability to differentiate between strains. Here we report the further development of this method, called fast ligation-mediated PCR (FLiP), which allows analysis of strains within one working day and starting from less than 1 ng of mycobacterial DNA or a crude cell lysate. Blinded analysis of a standard set of 131 M. tuberculosis complex and nontuberculous isolates showed the ability to differentiate 81 types among 90 M. tuberculosis complex isolates with 84 different IS6110 RFLP fingerprint patterns and detected 97% of the 31 duplicate samples. We suggest that FLiP can serve to rapidly detect chains of transmission prior to starting high-throughput analysis or standard IS6110 RFLP. It may as well serve as a secondary typing technique for other, non-IS6110-based methods. PMID- 16272497 TI - Evaluation of techniques for detection of carbapenem-resistant Klebsiella pneumoniae in stool surveillance cultures. AB - Screening for gastrointestinal colonization with multidrug-resistant nosocomial pathogens is an important component of infection control protocols. In the New York City region, carbapenem-resistant Klebsiella pneumoniae strains, which harbor the KPC carbapenem-hydrolyzing beta-lactamase, have rapidly emerged. The potential utility of screening medium, which involved using 10-mug imipenem disks, was investigated. The method of placing a sample from a fecal surveillance culture into broth containing an imipenem disk appeared to have the greatest sensitivity for detecting KPC-producing K. pneumoniae. Gastrointestinal colonization with two other carbapenem-resistant nosocomial pathogens, Pseudomonas aeruginosa and Acinetobacter baumannii, was also detected using this method. Placing fecal surveillance specimens into broth containing an imipenem disk is an easy method for screening samples for carbapenem-resistant nosocomial pathogens. PMID- 16272496 TI - Discriminatory power and reproducibility of novel DNA typing methods for Mycobacterium tuberculosis complex strains. AB - In recent years various novel DNA typing methods have been developed which are faster and easier to perform than the current internationally standardized IS6110 restriction fragment length polymorphism typing method. However, there has been no overview of the utility of these novel typing methods, and it is largely unknown how they compare to previously published methods. In this study, the discriminative power and reproducibility of nine recently described PCR-based typing methods for Mycobacterium tuberculosis were investigated using the strain collection of the interlaboratory study of Kremer et al. This strain collection contains 90 M. tuberculosis complex and 10 non-M. tuberculosis complex mycobacterial strains, as well as 31 duplicated DNA samples to assess reproducibility. The highest reproducibility was found with variable numbers of tandem repeat typing using mycobacterial interspersed repetitive units (MIRU VNTR) and fast ligation-mediated PCR (FLiP), followed by second-generation spoligotyping, ligation-mediated PCR (LM-PCR), VNTR typing using five repeat loci identified at the Queens University of Belfast (QUB VNTR), and the Amadio speciation PCR. Poor reproducibility was associated with fluorescent amplified fragment length polymorphism typing, which was performed in three different laboratories. The methods were ordered from highest discrimination to lowest by the Hunter-Gaston discriminative index as follows: QUB VNTR typing, MIRU VNTR typing, FLiP, LM-PCR, and spoligotyping. We conclude that both VNTR typing methods and FLiP typing are rapid, highly reliable, and discriminative epidemiological typing methods for M. tuberculosis and that VNTR typing is the epidemiological typing method of choice for the near future. PMID- 16272498 TI - Comparison of an automated repetitive sequence-based PCR microbial typing system to pulsed-field gel electrophoresis for analysis of outbreaks of methicillin resistant Staphylococcus aureus. AB - Rapid and sensitive methods for accurate strain delineation are essential for monitoring and preventing transmission of methicillin-resistant Staphylococcus aureus (MRSA). Pulsed-field gel electrophoresis (PFGE) has been the standard technique for strain typing most bacterial species including MRSA. The goal of this study was to compare the performance of the DiversiLab microbial typing system (Bacterial BarCodes, Inc., Houston, TX) (rep-PCR) to that of PFGE for typing MRSA isolates from five well-defined outbreaks. The DiversiLab rep-PCR assay is a rapid, semiautomated method based on PCR amplification of specific regions between noncoding repetitive sequences in the bacterial genome. rep-PCR was performed according to the manufacturer's recommendations, and the results were analyzed and dendrograms were generated using the DiversiLab analysis software (version 2.1.66 a). PFGE was performed and interpreted according to published procedures. rep-PCR results using similarity indices (SI) of 80%, 85%, and 90% were compared to PFGE analysis. In addition, intra- and interrun reproducibility was determined for rep-PCR. Overall, correct assignment to outbreak versus nonoutbreak clusters occurred for 91 of 109 isolates (85% agreement) when using a SI of 85%. For each specific outbreak, concordance between rep-PCR and PFGE ranged from 73% to 100%. There were 18 discrepant results (17%). Fourteen isolates were unique by PFGE, but they were placed in clusters by rep-PCR; the other 4 were placed in clusters different from those assigned by PFGE. Intra- and interrun reproducibility was excellent. Times to results were 12 to 24 h for rep-PCR compared to 2 to 4 days for PFGE. Rapid, standardized results and excellent reproducibility make rep-PCR a valuable tool for use in MRSA investigations. However, since rep-PCR was less discriminatory than PFGE, we recommend that it be used to screen isolates, followed by testing isolates which share the same rep-PCR pattern with a more sensitive method, such as PFGE or multilocus sequence typing. PMID- 16272499 TI - Sensitive EDTA-based microbiological assays for detection of metallo-{beta} lactamases in nonfermentative gram-negative bacteria. AB - The worldwide spread of metallo-beta-lactamase (MBL)-producing gram-negative bacilli represents a great concern nowadays. Sensitive assays for their specific detection are increasingly demanded to aid infection control and to prevent their dissemination. We have developed a novel microbiological assay employing crude bacterial extracts, designated EDTA-imipenem microbiological assay (EIM), to identify MBLs in nonfermentative gram-negative clinical strains. We also evaluated the ability of EIM to detect MBLs in comparison to those of other currently employed screening methods, such as the EDTA disk synergy test (EDS) with imipenem as a substrate and the Etest method. The sensitivities of EIM and Etest were similar (1 versus 0.92, respectively) and much higher than that of EDS (0.67). Moreover, both EIM and Etest displayed the maximum specificity. Modifications were introduced to EDS, including the simultaneous testing of three different beta-lactams (imipenem, meropenem, and ceftazidime) and two different EDTA concentrations. This resulted in a sensitivity improvement (0.92), albeit at a cost to its specificity. A simple strategy to accurately detect MBL producers is proposed; this strategy combines (i) an initial screening of the isolates by the extended EDS assay to select the potential candidates and (ii) confirmation of the true presence of MBL activity by EIM. PMID- 16272501 TI - Characterization of clonal complexity in tuberculosis by mycobacterial interspersed repetitive unit-variable-number tandem repeat typing. AB - In recent years, the application of molecular tools has shown us that clonal complexity in infection by Mycobacterium tuberculosis is not anecdotal. Exogenous reinfections, mixed infections, compartmentalization, and microevolution are different aspects of this issue. The detection and characterization of clonal variants of M. tuberculosis by standard genotyping methods is laborious and frequently requires expertise. Our aim was to evaluate a new genotyping PCR-based method for M. tuberculosis, mycobacterial interspersed repetitive unit-variable number tandem repeat typing (MIRU-VNTR), as a potential tool to simplify and optimize the clonal analysis of tuberculosis. MIRU-VNTR was able to detect mixed clonal variants in vitro, even for clones at low ratios (1:99). This technique was prospectively applied to search for cases infected by more than one clone. Clonal variants within the same host were detected in 3 out of 115 cases (2.6%), including cases with clones which were indistinguishable by restriction fragment length polymorphism or spoligotyping. In one case, coinfecting clonal variants differed in antibiotic susceptibilities. MIRU-VNTR was applied to cases with proven polyclonal infection, and it succeeded in detecting the coinfecting strains and proved useful in confirming cases of compartmentalized infection. MIRU-VNTR is a simple, rapid, and sensitive method which could facilitate and optimize the identification and characterization of clonal complexity in M. tuberculosis infection. PMID- 16272502 TI - Characteristics of Haemophilus influenzae type b responsible for meningitis in Poland from 1997 to 2004. AB - Two hundred forty-five H. influenzae isolates responsible for meningitis in Poland from 1997 to 2004 were studied. Among these, 233 (95.1%) belonged to serotype b (Hib), 2 belonged to serotype f, and 10 were noncapsulated. The relatedness of all isolates was evaluated by pulsed-field gel electrophoresis (PFGE), and selected representatives were evaluated by multilocus sequence typing. Resistance to ampicillin was identified in 34 (14.6%) of the Hib isolates and was associated with the production of beta-lactamase only. Except for four isolates nonsusceptible to chloramphenicol, all isolates were susceptible to cefotaxime, ciprofloxacin, and rifampin. The PFGE analysis divided the Hib isolates into five PFGE types; however, all of them were possibly related. The most common PFGE type, with 25 subtypes, was characteristic for 97.4% of the isolates. The most prevalent PFGE subtype found in our study was also the most common among the Hib isolates responsible for invasive disease in Italy and the Czech Republic and was found among isolates causing lower respiratory tract infections in Poland. The most prevalent sequence types (STs) in the studied group were ST6 and ST92. Four new STs were found: ST188, ST189, ST190, and ST268. Results of this study support the evidence that the genetic structure of encapsulated H. influenzae is clonal. The continuing high number of meningitis cases due to Hib in Poland underlines the need for mass vaccination against Hib in Poland. PMID- 16272500 TI - Specific and sensitive detection of Neisseria gonorrhoeae in clinical specimens by real-time PCR. AB - Early diagnosis of Neisseria gonorrhoeae infections is important with regard to patients' health and infectivity. We report the development of a specific and sensitive TaqMan assay for the detection of N. gonorrhoeae in clinical samples. The target sequence is a 76-bp fragment of the 5' untranslated region of the opa genes that encode opacity proteins. A panel of 448 well-defined N. gonorrhoeae isolates was used to evaluate and optimize the assay. The method employs two minor-groove binding probes, one of them recognizing a newly identified sequence in the opa genes. Testing a large panel of related and unrelated microorganisms revealed that other Neisseria strains and other microorganisms tested negative in the opa test. With a lower detection limit of one genome per reaction, the opa test appeared more sensitive than both the COBAS AMPLICOR (Roche Diagnostics Nederland BV, Almere, The Netherlands) and a LightCycler 16S rRNA test. Analysis of a panel of 122 COBAS AMPLICOR-positive samples revealed that 68% were negative in both the 16S rRNA test and the opa assay (confirming that the COBAS AMPLICOR test produces false positives), while 30% were positive in both assays. Three samples were opa positive and 16S rRNA negative, which may be due to the higher sensitivity of the opa assay. We conclude that the opa gene-based real-time amplification assay offers a sensitive, specific, semiquantitative, and reliable assay suitable for the detection of N. gonorrhoeae in clinical specimens and/or for confirmation of less specific tests. PMID- 16272503 TI - Direct detection and identification of Mycobacterium tuberculosis and Mycobacterium bovis in bovine samples by a novel nested PCR assay: correlation with conventional techniques. AB - Mycobacterium tuberculosis and M. bovis infect animals and humans. Their epidemiology in developed and developing countries differs, owing to differences in the implementation of preventive measures (World Health Organization, 1999). Identification and differentiation of these closely related mycobacterial species would help to determine the source, reservoirs of infection, and disease burden due to diverse mycobacterial pathogens. The utility of the hupB gene (Rv2986c in M. tuberculosis, or Mb3010c in M. bovis) to differentiate M. tuberculosis and M. bovis was evaluated by a PCR-restriction fragment length polymorphism (RFLP) assay with 56 characterized bovine isolates. The degree of concordance between the PCR-RFLP assay and the microbiological characterization was 99.0% (P < 0.001). A nested PCR (N-PCR) assay was developed, replacing the PCR-RFLP assay for direct detection of M. tuberculosis and M. bovis in bovine samples. The N-PCR products of M. tuberculosis and M. bovis corresponded to 116 and 89 bp, respectively. The detection limit of mycobacterial DNA by N-PCR was 50 fg, equivalent to five tubercle bacilli. M. tuberculosis and/or M. bovis was detected in 55.5% (105/189) of the samples by N-PCR, compared to 9.4% (18/189) by culture. The sensitivities of N-PCR and culture were 97.3 and 29.7, respectively, and their specificities were 22.2 and 77.7%, respectively. The percentages of animals or samples identified as infected with M. tuberculosis or M. bovis by N-PCR and culture reflected the clinical categorizations of the cattle (P of <0.05 to <0.01). Mixed infection by N-PCR was detected in 22 animals, whereas by culture mixed infection was detected in 1 animal. PMID- 16272504 TI - Detection of anti-hepatitis C virus effects of interferon and ribavirin by a sensitive replicon system. AB - Although combination therapy with interferon and ribavirin has improved the treatment for chronic hepatitis C virus (HCV) infection, the detailed anti-HCV effect of ribavirin in clinical concentrations remains uncertain. To detect the anti-HCV effect of ribavirin in lower concentrations, a sensitive and accurate assay system was developed using the reporter replicon system with an HCV genotype 2a subgenomic replicon (clone JFH-1) that exhibits robust replication in various cell lines. This reporter replicon was generated by introducing the luciferase reporter gene (instead of the neomycin resistance gene) into the subgenomic JFH-1 replicon. To assess the replication of this reporter replicon, luciferase activity was measured serially up to day 3 after transient transfection of Huh7 cells. The luciferase activity increased exponentially over the time course of the experiment. After adjustment for transfection efficiency and transfected cell viability, the impacts of interferon and ribavirin were determined. The administration of interferon and ribavirin resulted in dose dependent suppression of replicon RNA replications. The 50% inhibitory concentration of interferon and ribavirin was 1.80 IU/ml and 3.70 microg/ml, respectively. In clinical concentrations, replications were reduced to 0.09% and 53.74% by interferon (100 IU/ml) and ribavirin (3 microg/ml), respectively. Combination use of ribavirin and interferon enhanced the anti-HCV effect of interferon by 1.46- to 1.62-fold. In conclusion, we developed an accurate and sensitive replicon system, and the antivirus effect of interferon and ribavirin was easily detected within their clinical concentrations by this replicon system. This system will provide a powerful tool for screening new antiviral compounds against HCV. PMID- 16272505 TI - Rapid and accurate genotyping of YMDD motif variants in the hepatitis B virus genome by an improved reverse dot blot method. AB - By using the "flow-through hybridization" principle, we developed a new, rapid and accurate reverse dot blot (RDB) method to detect lamivudine resistance associated YMDD motif variants in hepatitis B virus (HBV) genome. The improved RDB method was very fast at simultaneously detecting HBV YMDD wild-type and mutant motifs. In a blind analysis, 100 samples previously genotyped by DNA clonal sequencing analysis were used to evaluate the sensitivity and specificity of this assay. Conventional restriction fragment length polymorphism (RFLP) data were also used to test our method. In blind experiments, our improved RDB method had an accuracy and specificity of 100%, which was much higher than RFLP, which had an accuracy and specificity of only 83.0%. In clinical detection practice, 49 patients highly suspected of lamivudine resistance were successfully diagnosed by this method. Our improved RDB assay is a simple, rapid, cheap, semiautomatic, accurate, sensitive, and contamination-proof method of detecting lamivudine resistance-associated mutants in the human hepatitis B virus genome. PMID- 16272506 TI - Use of resequencing oligonucleotide microarrays for identification of Streptococcus pyogenes and associated antibiotic resistance determinants. AB - Group A streptococci (GAS) are responsible for a wide variety of human infections associated with considerable morbidity and mortality. Ever since the first systematic effort by Lancefield to group Streptococcus species by M protein variants, the detection and characterization of Streptococcus by different methods have been an evolving process. The ideal assay for GAS identification not only would provide quick and accurate diagnostic results but also would reveal antibiotic resistance patterns and genotype information, aiding not only in treatment but in epidemiologic assessment as well. The oligonucleotide microarray is a promising new technology which could potentially address this need. In this study, we evaluated the usefulness of oligonucleotide resequencing microarrays for identifying GAS and its associated antibiotic resistance markers. We demonstrated an assay platform that combines the use of resequencing DNA microarrays with either random nucleic acid amplification or multiplex PCR for GAS detection. When detecting Streptococcus pyogenes from coded clinical samples, this approach demonstrated an excellent concordance with a more established culture method. To this end, we showed the potential of resequencing microarrays for efficient and accurate detection of GAS and its associated antibiotic resistance markers with the benefit of sequencing information from microarray analysis. PMID- 16272507 TI - Sensitive phenotypic detection of minor drug-resistant human immunodeficiency virus type 1 reverse transcriptase variants. AB - Detection of drug-resistant variants is important for the clinical management of human immunodeficiency virus type 1 (HIV-1) infection and for studies on the evolution of drug resistance. Here we show that hybrid elements composed of the Saccharomyces cerevisiae retrotransposon Ty1 and the reverse transcriptase (RT) of HIV-1 are useful tools for detecting, monitoring, and isolating drug-resistant reverse transcriptases. This sensitive phenotypic assay is able to detect nonnucleoside reverse transcriptase inhibitor-resistant RT domains derived from mixtures of infectious molecular clones of HIV-1 in plasma and from clinical samples when the variants comprise as little as 0.3 to 1% of the virus population. Our assay can characterize the activities and drug susceptibilities of both known and novel reverse transcriptase variants and should prove useful in studies of the evolution and clinical significance of minor drug-resistant viral variants. PMID- 16272508 TI - Campylobacter coli in swine production: antimicrobial resistance mechanisms and molecular epidemiology. AB - The aim of this study was to determine antimicrobial resistance, to evaluate and compare the use of two genotyping methods for molecular epidemiology purposes, and to determine the genotypic diversity of Campylobacter coli of porcine origin. A total of 100 C. coli isolates from swine were tested for susceptibility to six antimicrobials using the agar dilution method and genotyped using two high resolution fingerprinting approaches: multilocus sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE). Evaluation of the methods was based on their resistance patterns, discriminatory indexes (DI), high test throughputs, costs, and turnaround times. Resistance to erythromycin and tetracycline was the most common. Both genotypic methods were found to have high discriminatory power, although MLST had a higher DI (0.936) than PFGE (DI = 0.889). It also had a higher throughput than PFGE. Isolates were clustered into 27 groups by MLST compared to 11 by PFGE. MLST was able to further discriminate the isolates grouped under the same cluster by PFGE. Out of the 65 MLST sequence types (STs) identified among the total isolates, 50 were reported for the first time. Most STs were found to be specific to the farm (n = 38) and to slaughter (n = 22). Resistance against tetracycline and erythromycin was encoded by the tet(O) gene and a A2075G point mutation in the 23S rRNA gene, respectively. A high ciprofloxacin MIC (>64 microg/liter) was conferred by a point mutation in the gyrA gene. The weak clonal structure of the C. coli population among swine was further highlighted by the index of association value of 0.293. The findings of this study indicate that multidrug-resistant diverse C. coli strains exhibiting resistance to ciprofloxacin and erythromycin are concerning, since these are the drugs of choice for treating invasive campylobacteriosis cases in humans. PMID- 16272509 TI - Distribution of tccP in clinical enterohemorrhagic and enteropathogenic Escherichia coli isolates. AB - Enterohemorrhagic Escherichia coli (EHEC) and enteropathogenic E. coli (EPEC) are diarrheagenic pathogens that colonize the gut through the formation of attaching and effacing lesions, which depend on the translocation of effector proteins via a locus of enterocyte effacement-encoded type III secretion system. Recently, two effector proteins, EspJ and TccP, which are encoded by adjacent genes on prophage CP-933U in EHEC O157:H7, have been identified. TccP consists of a unique N terminus region and several proline-rich domains. In this project we determined the distribution of tccP in O157:H7, in non-O157 EHEC, and in typical and atypical EPEC isolates. All the EHEC O157:H7 strains tested were tccP(+). Unexpectedly, tccP was also found in non-O157 EHEC, and in typical and atypical EPEC isolates, particularly in strains belonging to serogroups O26 (EHEC), O119 (typical EPEC), and O55 (atypical EPEC). We recorded some variation in the length of tccP, which reflects diversity in the number of the proline-rich repeats. These results show the existence of a class of "attaching and effacing" pathogens which express a combination of EPEC and EHEC virulence determinants. PMID- 16272510 TI - Defining the normal bacterial flora of the oral cavity. AB - More than 700 bacterial species or phylotypes, of which over 50% have not been cultivated, have been detected in the oral cavity. Our purposes were (i) to utilize culture-independent molecular techniques to extend our knowledge on the breadth of bacterial diversity in the healthy human oral cavity, including not yet-cultivated bacteria species, and (ii) to determine the site and subject specificity of bacterial colonization. Nine sites from five clinically healthy subjects were analyzed. Sites included tongue dorsum, lateral sides of tongue, buccal epithelium, hard palate, soft palate, supragingival plaque of tooth surfaces, subgingival plaque, maxillary anterior vestibule, and tonsils. 16S rRNA genes from sample DNA were amplified, cloned, and transformed into Escherichia coli. Sequences of 16S rRNA genes were used to determine species identity or closest relatives. In 2,589 clones, 141 predominant species were detected, of which over 60% have not been cultivated. Thirteen new phylotypes were identified. Species common to all sites belonged to the genera Gemella, Granulicatella, Streptococcus, and Veillonella. While some species were subject specific and detected in most sites, other species were site specific. Most sites possessed 20 to 30 different predominant species, and the number of predominant species from all nine sites per individual ranged from 34 to 72. Species typically associated with periodontitis and caries were not detected. There is a distinctive predominant bacterial flora of the healthy oral cavity that is highly diverse and site and subject specific. It is important to fully define the human microflora of the healthy oral cavity before we can understand the role of bacteria in oral disease. PMID- 16272511 TI - Molecular epidemiology of clinical Cryptococcus neoformans strains from India. AB - Little is known about the molecular epidemiology of the human pathogenic fungus Cryptococcus neoformans in India, a country now in the midst of an epidemic of AIDS-related cryptococcosis. We studied 57 clinical isolates from several regions in India, of which 51 were C. neoformans var. grubii, 1 was C. neoformans var. neoformans, and 5 were C. neoformans var. gattii. This strain set included 18 additional sequential isolates from 14 patients. Strains were characterized phenotypically by measuring the polysaccharide capsule and by determining the MICs of standard antifungals. Molecular typing was performed by a PCR-based method using the minisatellite-specific core sequence (M13), by electrophoretic karyotyping, by restriction fragment length polymorphisms with the C. neoformans transposon 1 (TCN-1), and by URA5 DNA sequence analysis. Overall, Indian isolates were less heterogeneous than isolates from other regions and included a subset that clustered into one group based on URA5 DNA sequence analysis. In summary, our results demonstrate (i) differences in genetic diversity of C. neoformans isolates from India compared to isolates from other regions in the world; (ii) that DNA typing with the TCN-1 probe can adequately distinguish C. neoformans var. grubii strains; (iii) that TCN-1 sequences are absent in many C. neoformans var. gattii strains, supporting previous studies indicating that these strains have a limited geographical dispersal; and (iv) that human cryptococcal infection can be associated with microevolution of the infecting strain and by simultaneous coinfection with two distinct C. neoformans strains. PMID- 16272512 TI - PCR analysis of egyptian respiratory adenovirus isolates, including identification of species, serotypes, and coinfections. AB - Eighty-eight adenovirus (Ad) isolates and associated clinical data were collected from walk-in patients with influenza-like illness in Egypt during routine influenza surveillance from 1999 through 2002. Respiratory Ad distributions are geographically variable, and serotype prevalence has not been previously characterized in this region. Serotype identity is clinically relevant because it predicts vaccine efficacy and correlates strongly with both clinical presentation and epidemiological pattern. Species and serotype identities were determined using several well-validated multiplex PCR protocols culled from the literature and supplemented with a few novel primer sets designed to identify rare types. The isolates included common species B1 serotypes (Ad3 and Ad7), common species C serotypes (Ad1, Ad2, and Ad5), the less common species B2 serotype Ad11, and three isolates of the rare species B1 serotype Ad16. Two isolates that appear to be variant Ad16 were also identified. Fifteen coinfections of multiple adenoviral types, primarily AdB/AdC and Ad3/Ad7 dual infections, were detected. The majority of these were verified using redundant PCR tests targeted at multiple genes. PCR is able to resolve coinfections, in contrast to traditional serum neutralization tests. PCR is also comparatively rapid and requires very little equipment. Application of the method allowed an inclusive determination of the serotypes found in the Egyptian respiratory sample set and demonstrated that coinfections are common and may play a previously unrecognized role in adenovirus pathogenesis, evolution, and epidemiology. In particular, coinfections may influence adenoviral evolution, as interserotypic recombination has been identified as a source of emerging strains. PMID- 16272513 TI - Microbial risk indicators of early childhood caries. AB - The aim of this study was to use molecular identification methods, such as 16S RNA gene sequence and reverse-capture checkerboard hybridization, for identification of the bacteria associated with dental caries and with dental health in a subset of 204 twins aged 1.5 to 7 years old. A total of 448 plaque samples (118 collected from caries-free subjects and 330 from caries-active subjects) were used for analysis. We compared the bacteria found in biofilms of children exhibiting severe dental caries, with different degrees of lesion severity, with those found in biofilms of caries-free children. A panel of 82 bacterial species was selected, and a PCR-based reverse-capture checkerboard method was used for detection. A simple univariate test was used to determine the overabundance and underabundance of bacterial species in the diseased and in the healthy groups. Features identified with this univariate test were used to construct a probabilistic disease prediction model. Furthermore, a method for the analysis of global patterns of gene expression was performed to permit simultaneous analysis of the abundance of significant species by allowing cross bacterial comparisons of abundance profiles between caries-active and caries-free subjects. Our results suggested that global patterns of microbial abundance in this population are very distinctive. The top bacterial species found to be overabundant in the caries-active group were Actinomyces sp. strain B19SC, Streptococcus mutans, and Lactobacillus spp., which exhibited an inverse relationship to beneficial bacterial species, such as Streptococcus parasanguinis, Abiotrophia defectiva, Streptococcus mitis, Streptococcus oralis, and Streptococcus sanguinis. PMID- 16272515 TI - Supplementation of CHROMagar Candida medium with Pal's medium for rapid identification of Candida dubliniensis. AB - CHROMagar Candida medium is used for the isolation and identification of Candida species, but it does not differentiate Candida albicans from Candida dubliniensis. This differentiation can be achieved by using Pal's agar, which cannot be used in primary isolation. We have combined both media to obtain a new medium that can be used for the isolation and identification of C. dubliniensis in primary cultures. PMID- 16272514 TI - Human infection with an avian H9N2 influenza A virus in Hong Kong in 2003. AB - Avian H9N2 influenza A virus has caused repeated human infections in Asia since 1998. Here we report that an H9N2 influenza virus infected a 5-year-old child in Hong Kong in 2003. To identify the possible source of the infection, the human isolate and other H9N2 influenza viruses isolated from Hong Kong poultry markets from January to October 2003 were genetically and antigenically characterized. The findings of this study show that the human H9N2 influenza virus, A/Hong Kong/2108/03, is of purely avian origin and is closely related to some viruses circulating in poultry in the markets of Hong Kong. The continued presence of H9N2 influenza viruses in poultry markets in southern China increases the likelihood of avian-to-human interspecies transmission. PMID- 16272516 TI - Use of a real-time PCR TaqMan assay for rapid identification and differentiation of Burkholderia pseudomallei and Burkholderia mallei. AB - A TaqMan allelic-discrimination assay designed around a synonymous single nucleotide polymorphism was used to genotype Burkholderia pseudomallei and Burkholderia mallei isolates. The assay rapidly identifies and discriminates between these two highly pathogenic bacteria and does not cross-react with genetic near neighbors, such as Burkholderia thailandensis and Burkholderia cepacia. PMID- 16272517 TI - New quinolone resistance phenomenon in Salmonella enterica: nalidixic acid susceptible isolates with reduced fluoroquinolone susceptibility. AB - We describe the emergence of a new quinolone resistance pattern in Salmonella enterica isolates from Southeast Asia. These isolates are susceptible to nalidixic acid but exhibit reduced susceptibility to ciprofloxacin. The increase of such strains may threaten the value of the nalidixic acid disk test to screen for reduced fluoroquinolone susceptibility in salmonellas. PMID- 16272518 TI - Distribution of strain families of Mycobacterium tuberculosis causing pulmonary and extrapulmonary disease in hospitalized children in Cape Town, South Africa. AB - We studied the association between strain family and extrapulmonary tuberculosis among 285 children presenting to a pediatric hospital. Extrapulmonary disease occurred in 56% of children without known human immunodeficiency virus infection, with meningitis accounting for 22% of the cases. Two strain families, LAM3/F11 and W-Beijing, predominated; but there was no overall association with extrapulmonary disease. PMID- 16272520 TI - Use of four dengue virus antigens for determination of dengue immune status by enzyme-linked immunosorbent assay of immunoglobulin G avidity. AB - We used an enzyme-linked immunosorbent assay (ELISA) of immunoglobulin G avidity to determine the dengue immune status of 105 pairs of serum samples from patients infected with dengue virus. This study shows that a simple avidity test, for which only one acute-phase serum sample is required, is potentially more useful than the hemagglutination inhibition test for the discrimination of primary from secondary dengue virus infection, whatever the type of dengue antigen used. PMID- 16272519 TI - Comparison of the RSV respi-strip with direct fluorescent-antigen detection for diagnosis of respiratory syncytial virus infection in pediatric patients. AB - The RSV Respi-Strip was compared to the SimulFluor respiratory screen for detecting respiratory syncytial virus in nasopharyngeal aspirates from pediatric patients. Of samples tested, 115/239 (49%) were positive by direct fluorescent antigen detection. The sensitivity and specificity of the RSV Respi-Strip were 92% (95% confidence interval [CI], 86% to 96%) and 98% (95% CI, 94% to 100%), respectively, with a diagnostic efficiency of 95%. PMID- 16272522 TI - Detection of a bla(SHV) extended-spectrum {beta}-lactamase in Salmonella enterica serovar Newport MDR-AmpC. AB - Salmonella enterica serovar Newport MDR-AmpC expressing TEM-1b and extended spectrum beta-lactamase SHV-12 was isolated from affected animals during an outbreak of salmonellosis that led to a 3-month closure of one of the largest equine hospitals in the United States. PMID- 16272521 TI - Phylogenetic analysis of the p24-p7 region of the human immunodeficiency virus type 1 gag gene to determine subtype distribution among female sex workers in Calcutta, India. AB - Human immunodeficiency virus type 1 (HIV-1) subtype C, based on the envelope region, has been reported to be predominant in India. We sequenced the p24-p7 gag region from 51 HIV-1 seropositive female sex workers in Calcutta, India, for more detailed molecular characterization. Subtype C was found to be prevalent, although no strong monophyletic cluster was observed. PMID- 16272523 TI - Increased rate of DNA recovery from United Kingdom epidemic Clostridium difficile PCR ribotype 1 strains stored cryogenically. AB - We noted that some Clostridium difficile isolates are nonrecoverable after frozen storage and so used molecular typing analysis to characterize DNA from these strains. The recovery rate of C. difficile PCR ribotype 1 was statistically significantly greater than that of other strains. This observation has implications for C. difficile epidemiological studies. PMID- 16272524 TI - Epidemiology and molecular analysis of intestinal colonization by vancomycin resistant enterococci in greek hospitals. AB - From 1,246 specimens collected from 13 Greek hospitals, 266 vancomycin-resistant enterococci strains were isolated from 255 patients (20.5%). The VanA phenotype was present in 82 (30.8%) strains, the VanB phenotype in 17 (6.4%) strains, the VanC1 phenotype in 152 (57.1%) strains, and the VanC2/C3 phenotypes in 15 (5.6%) strains. When only VanA and VanB phenotypes were considered, the overall prevalence was 7.5%. Eighty-six isolates exhibiting the VanA or VanB phenotype were analyzed by pulsed-field gel electrophoresis (PFGE), and 46 PFGE groups were found. PMID- 16272525 TI - Emergence of a Streptococcus pneumoniae clinical isolate highly resistant to telithromycin and fluoroquinolones. AB - Streptococcus pneumoniae is a major pathogen causing community-acquired pneumonia and acute bronchitis. Macrolides, fluoroquinolones (FQs), and, recently, telithromycin (TEL) constitute primary therapeutic options, and rare cases of resistance have been reported. In this report, we describe the emergence of an S. pneumoniae clinical isolate with high-level TEL resistance (MIC, 256 microg/ml) and simultaneous resistance to FQs. Ongoing studies are oriented to elucidate the precise mechanism of resistance to TEL. PMID- 16272526 TI - Broth microdilution susceptibility testing of Brucella species: quality control limits for ten antimicrobial agents against three standard quality control strains. AB - Brucella broth without supplementation is the recommended medium for broth microdilution susceptibility tests of Brucella abortus, B. melitensis, and B. suis. Based on an eight-laboratory collaborative study using a pH-adjusted modification of this medium, we propose MIC quality control ranges for three control strains against 10 antimicrobials that are potentially efficacious for treating infections caused by these agents of bioterrorism. PMID- 16272527 TI - Evaluation of a fully automated system (RAISUS) for rapid identification and antimicrobial susceptibility testing of Staphylococci. AB - RAISUS is a system for rapid bacterial identification and antimicrobial susceptibility testing. RAISUS and VITEK showed 97.8% and 75.9% agreement in identification of 45 Staphylococcus aureus strains and 58 coagulase-negative staphylococci (CoNS), respectively, and RAISUS and CLSI (formerly NCCLS) methods showed 87.2% and 87.9% agreement in the MICs for S. aureus and CoNS, respectively. RAISUS provided these data within 3.75 h, suggesting its utility for clinical bacteriological laboratories. PMID- 16272528 TI - Laryngeal scleroma associated with Klebsiella pneumoniae subsp. ozaenae. AB - Klebsiella pneumoniae subsp. ozaenae was isolated from the pharynx of a woman with laryngeal scleroma. K. pneumoniae subsp. ozaenae is rarely isolated from clinical infections and has never been reported in laryngeal scleroma, which is usually caused by K. pneumoniae subsp. rhinoscleromatis. PMID- 16272529 TI - Three nonfatal cases of neonatal adenovirus infection. AB - Neonatal adenovirus infection is considered a rare and fatal disease. Three nonfatal neonatal adenovirus infections manifesting as conjunctivitis or conjunctivitis with other nonspecific symptoms are described. Adenovirus DNA was detected by PCR in eye swabs from two patients and in both cerebrospinal fluid and eye swabs in the third patient. PMID- 16272530 TI - Group A streptococcal meningitis in a pediatric patient following cochlear implantation: report of the first case and review of the literature. AB - The Food and Drug Administration published a public health warning on the association of bacterial meningitis and cochlear implants in June 2002. This article reports the first case of group A streptococcal (GAS) meningitis in a cochlear-implanted patient, followed by a review on cochlear implantation and GAS meningitis. PMID- 16272531 TI - Molecular identification of Rhizomucor pusillus as a cause of sinus-orbital zygomycosis in a patient with acute myelogenous leukemia. AB - Sinus-orbital zygomycosis caused by Rhizomucor pusillus in a patient with acute myelogenous leukemia is described. Identification was achieved by sequencing of the internal transcribed spacer (ITS) regions of the rRNA gene and by expression of zygospores in mating. This report highlights the value of ITS sequencing as a diagnostic tool for the identification of R. pusillus and expands the understanding of infection types caused by this zygomycete. PMID- 16272532 TI - Non-culture-based analysis of bacterial populations from patients with chronic rhinosinusitis. AB - Middle meatus aspirates from patients with chronic rhinosinusitis were analyzed by bacterial culture, denaturing gradient gel electrophoresis (DGGE), and antibiotic sensitivity techniques. DGGE detected a greater bacterial diversity than culture methods. Although resistance to antibiotics was low, there was evidence of changes in the composition of the bacterial microbiota over time, and the presence of noncultured bacteria was demonstrated. PMID- 16272533 TI - Intraabdominal zygomycosis caused by Syncephalastrum racemosum infection successfully treated with partial surgical debridement and high-dose amphotericin B lipid complex. AB - Invasive zygomycosis rarely complicates trauma. We describe the first recorded case of invasive infection of the anterior abdominal wall and omentum with the zygomycete Syncephalastrum racemosum, which was successfully treated with partial surgical debridement and amphotericin B lipid complex. PMID- 16272535 TI - Commercial mushrooms and bean sprouts are a source of Pseudomonas aeruginosa. PMID- 16272534 TI - The role of cryptococcal antigen assay in diagnosis and monitoring of cryptococcal meningitis. PMID- 16272536 TI - Vitek 2 automated identification system and Kocuria kristinae. PMID- 16272537 TI - Review of avian quarantine arrangements to report within a month. PMID- 16272543 TI - Observations on salpingitis, peritonitis and salpingoperitonitis in a layer breeder flock. AB - A flock of 13,951 hens and 1379 cockerels was monitored from 26 to 58 weeks of age for the complex of salpingitis, peritonitis and salpingoperitonitis (sps). Two hundred and forty-three hens (78 per cent of the hens that died) were examined postmortem, and sps was recognised by gross examination for inflammatory exudate, in the body cavity or oviduct in 111 (46 per cent) of them. Salpingoperitonitis was the most common form, followed by salpingitis and then peritonitis. There were acute and chronic cases in all three conditions, but only in peritonitis were acute cases more common than chronic cases. Seventeen birds that had died of sps were cultured for aerobic bacteria within 12 hours of death. Escherichia coli was recovered from a variety of tissues from all of them, and other bacteria, including staphylococci, Mannheimia haemolytica and Streptococcus bovis, were isolated from a few carcases, either alone or together with E coli. Relatively few isolations of E coli were made from normal hens cultured 48, 72 and 96 hours after death. PMID- 16272544 TI - Emergence of fasciolosis in cattle in East Anglia. AB - Liver fluke (Fasciola hepatica) infection caused weight loss, diarrhoea, decreased milk yield and occasionally death in cattle in East Anglia during the winters of 2001 to 2003. The condition had previously been limited mainly to stock imported into this part of Britain from endemically infected areas. In composite faecal samples collected by 16 farm animal veterinary practices in Norfolk, Suffolk and Essex, fluke eggs were found in 15 (28.8 per cent) of 52 previously unaffected suckler herds and 10 (16.7 per cent) of 60 dairy herds. Antibodies to F hepatica were detected by elisa in 32 (53.3 per cent) of the bulk milk samples from these 60 dairy herds, including the 10 in which fluke eggs were found. The emergence of fasciolosis in East Anglia was attributed to recent higher summer rainfall, which favoured the intermediate snail host Lymnaea truncatula and the free-living stages of F hepatica, the increased influx of sheep from endemic fluke areas for seasonal grazing, and the wetter grazing conditions associated with the Environmentally Sensitive Area scheme. PMID- 16272545 TI - Field evaluation of the efficacy of fenbendazole in captive wild ruminants. AB - The efficacy of in-feed fenbendazole at a dose rate of 7.5 mg/kg bodyweight for three consecutive days was assessed in five Arabian oryx (Oryx leucoryx), six scimitar-horned oryx (Oryx dammah), 14 slender-horned gazelles (Gazella leptoceros), eight Soay sheep (Ovis aries aries soay), 13 alpine ibex (Capra ibex ibex), six red deer (Cervus elaphus hippelaphus) and 11 Nelson's elk (Cervus elaphus nelsoni) kept in five herds in a zoo. The efficacy was assessed by means of repeated faecal egg count reduction (fecr) tests and in vitro egg hatch assays. Fenbendazole was highly effective against nematodes in five of the seven species, consistently reducing egg shedding by more than 90 per cent. In the egg hatch assays of the five herds, 50 per cent inhibition of hatching (ld50) was observed at a concentration of thiabendazole below 0.1 microg/ml. In the Arabian oryx and alpine ibex the efficacy of fenbendazole was less than 90 per cent, and the ld50 in the egg hatch assays was between 0.1 and 0.2 microg/ml thiabendazole. PMID- 16272546 TI - Verminous pneumonia due to Filaroides hirthi in a Scottish terrier in Spain. AB - A previously healthy Scottish terrier developed verminous pneumonia due to Filaroides hirthi. The disease was diagnosed on the basis of cytology of a transtracheal wash, which contained 17 per cent eosinophils and a large number of parasite larvae. Treatment with 50 mg/kg bodyweight fenbendazole daily for three weeks eliminated the parasites and improved the clinical condition of the dog. PMID- 16272547 TI - Serum catecholamine levels after halothane and isoflurane anaesthesia in rabbits. PMID- 16272548 TI - Recovery of Dictyocaulus species from the lungs of a Kashmir red deer (Cervus elaphus hanglu). PMID- 16272549 TI - Intraperitoneal actinomycosis in a cat. PMID- 16272552 TI - Avian influenza. PMID- 16272553 TI - Designation of approved veterinarians. PMID- 16272554 TI - Canine influenza virus. PMID- 16272555 TI - New medicines regulations. PMID- 16272556 TI - Doctor, doctor. PMID- 16272557 TI - Study on portosystemic shunts in dogs. PMID- 16272559 TI - Designation of 'new' mite species. PMID- 16272560 TI - Large scale protein profiling by combination of protein fractionation and multidimensional protein identification technology (MudPIT). AB - In the past decade, shotgun proteomic analysis has been utilized extensively to answer complex biological questions. New challenges arise in large scale proteomic profiling when dealing with complex biological mixtures such as the mammalian cell lysate. In this study, we explored the approach of protein separation prior to the shotgun multidimensional protein identification technology (MudPIT) analysis. We fractionated the mammalian cancer cell lysate using the PF 2D ProteomeLab system and analyzed the distribution of molecular weight, isoelectric point, and cellular localization of the eluted proteins. As a result, we were able to reduce sample complexity by protein fractionation and increase the possibility of detecting proteins with lower abundance in the complex protein mixture. PMID- 16272561 TI - The turnover kinetics of major histocompatibility complex peptides of human cancer cells. AB - Peptides presented by the major histocompatibility complex (MHC) are derived from the degradation of cellular proteins. Thus, the repertoire of these peptides (the MHC peptidome) should correlate better with the cellular protein degradation scheme (the degradome) than with the cellular proteome. To test the validity of this statement and to determine whether the majority of MHC peptides are derived from short lived proteins, from defective ribosome products, or from regular long lived cellular proteins we analyzed in parallel the turnover kinetics of both MHC peptides and cellular proteins in the same cancer cells. The analysis was performed by pulse-chase experiments based on stable isotope labeling in tissue culture followed by capillary chromatography and tandem mass spectrometry. Indeed only a limited correlation was observed between the proteome and the MHC peptidome observed in the same cells. Moreover a detailed analysis of the turnover kinetics of the MHC peptides helped to assign their origin to normal, to short lived or long lived proteins, or to the defective ribosome products. Furthermore the analysis of the MHC peptides turnover kinetics helped to direct attention to abnormalities in the degradation schemes of their source proteins. These observations can be extended to search for cancer-related abnormalities in protein degradation, including those that lead to loss of tumor suppressors and cell cycle regulatory proteins. PMID- 16272562 TI - Structural and functional insights into the interaction of echoviruses and decay accelerating factor. AB - Many enteroviruses bind to the complement control protein decay-accelerating factor (DAF) to facilitate cell entry. We present here a structure for echovirus (EV) type 12 bound to DAF using cryo-negative stain transmission electron microscopy and three-dimensional image reconstruction to 16-A resolution, which we interpreted using the atomic structures of EV11 and DAF. DAF binds to a hypervariable region of the capsid close to the 2-fold symmetry axes in an interaction that involves mostly the short consensus repeat 3 domain of DAF and the capsid protein VP2. A bulge in the density for the short consensus repeat 3 domain suggests that a loop at residues 174-180 rearranges to prevent steric collision between closely packed molecules at the 2-fold symmetry axes. Detailed analysis of receptor interactions between a variety of echoviruses and DAF using surface plasmon resonance and comparison of this structure (and our previous work; Bhella, D., Goodfellow, I. G., Roversi, P., Pettigrew, D., Chaudhry, Y., Evans, D. J., and Lea, S. M. (2004) J. Biol. Chem. 279, 8325-8332) with reconstructions published for EV7 bound to DAF support major differences in receptor recognition among these viruses. However, comparison of the electron density for the two virus.receptor complexes (rather than comparisons of the pseudo-atomic models derived from fitting the coordinates into these densities) suggests that the dramatic differences in interaction affinities/specificities may arise from relatively subtle structural differences rather than from large scale repositioning of the receptor with respect to the virus surface. PMID- 16272563 TI - Exendin-4 uses Irs2 signaling to mediate pancreatic beta cell growth and function. AB - The insulin receptor substrate 2 (Irs2) branch of the insulin/insulin-like growth factor-signaling cascade prevents diabetes in mice because it promotes beta cell replication, function, and survival, especially during metabolic stress. Because exendin-4 (Ex4), a long acting glucagon-like peptide 1 receptor agonist, has similar effects upon beta cells in rodents and humans, we investigated whether Irs2 signaling was required for Ex4 action in isolated beta cells and in Irs2(-/ ) mice. Ex4 increased cAMP levels in human islets and Min6 cells, which promoted Irs2 expression and stimulated Akt phosphorylation. In wild type mice Ex4 administered continuously for 28 days increased beta cell mass 2-fold. By contrast, Ex4 failed to arrest the progressive beta cell loss in Irs2(-/-) mice, which culminated in fatal diabetes; however, Ex4 delayed the progression of diabetes by 3 weeks by promoting insulin secretion from the remaining islets. We conclude that some short term therapeutic effects of glucagon-like peptide 1 receptor agonists can be independent of Irs2, but its long term effects upon beta cell growth and survival are mediated by the Irs2 branch of the insulin/insulin like growth factor signaling cascade. PMID- 16272565 TI - The properties of rabbit alpha1-macroglobulin upon activation are distinct from those of rabbit and human alpha2-macroglobulins. AB - We have characterized native and activated forms of rabbit alpha1M and compared them to rabbit and human alpha2M. Similar to human alpha2M, rabbit alpha1M is a tetramer associated via disulfide bonds and non-covalent interactions that exhibits autolysis into two fragments when heated. Like human alpha2M, rabbit alpha1M is cleaved by trypsin at one site; however, rabbit alpha1M shares characteristics with rabbit alpha2M that are different from the properties of human alpha2M. Amine or trypsin treatment of rabbit alpha-macroglobulins does not result in a significant conformational change or cleavage of four thiolester bonds. Full thiolester cleavage is only observed for rabbit alpha1M after exposure to both trypsin and a small amine. Additionally, amine-treated rabbit alpha-macroglobulins retain trypsin inhibitory potential and do not fully shield bound proteinases. Methylamine and trypsin treatment of rabbit alpha1M results in two dissimilar conformations that display differing exposure of the receptor recognition site. While ammonia- and methylamine-modified rabbit alpha1M bind to macrophages with similar affinity to that of human alpha2M, trypsin-treated rabbit alpha1M exhibits dramatically lower affinity. This suggests that rabbit alpha1M may not play the same proteinase-inhibiting physiological role as human alpha2M. PMID- 16272564 TI - The fasting-induced adipose factor/angiopoietin-like protein 4 is physically associated with lipoproteins and governs plasma lipid levels and adiposity. AB - Proteins secreted from adipose tissue are increasingly recognized to play an important role in the regulation of glucose metabolism. However, much less is known about their effect on lipid metabolism. The fasting-induced adipose factor (FIAF/angiopoietin-like protein 4/peroxisome proliferator-activated receptor gamma angiopoietin-related protein) was previously identified as a target of hypolipidemic fibrate drugs and insulin-sensitizing thiazolidinediones. Using transgenic mice that mildly overexpress FIAF in peripheral tissues we show that FIAF is an extremely powerful regulator of lipid metabolism and adiposity. FIAF overexpression caused a 50% reduction in adipose tissue weight, partly by stimulating fatty acid oxidation and uncoupling in fat. In addition, FIAF overexpression increased plasma levels of triglycerides, free fatty acids, glycerol, total cholesterol, and high density lipoprotein (HDL)-cholesterol. Functional tests indicated that FIAF overexpression severely impaired plasma triglyceride clearance but had no effect on very low density lipoprotein production. The effects of FIAF overexpression were amplified by a high fat diet, resulting in markedly elevated plasma and liver triglycerides, plasma free fatty acids, and plasma glycerol levels, and impaired glucose tolerance in FIAF transgenic mice fed a high fat diet. Remarkably, in mice the full-length form of FIAF was physically associated with HDL, whereas truncated FIAF was associated with low density lipoprotein. In human both full-length and truncated FIAF were associated with HDL. The composite data suggest that via physical association with plasma lipoproteins, FIAF acts as a powerful signal from fat and other tissues to prevent fat storage and stimulate fat mobilization. Our data indicate that disturbances in FIAF signaling might be involved in dyslipidemia. PMID- 16272566 TI - Regulation of cell adhesion and type VII collagen binding by the beta3 chain short arm of laminin-5: effect of its proteolytic cleavage. AB - The basement membrane protein laminin-5 (Lm5), a heterotrimer of alpha3 (or alpha3A), beta3, and gamma2 chains, regulates cellular adhesion and motility. Here we examined the proteolysis and biological function of the laminin beta3 chain. First, we found that the beta3 chain of Lm5 is cleaved at its N-terminal, short arm by an endogenous proteinase(s) in normal human keratinocytes and some other cell lines. To examine the effect of beta3 chain cleavage, we expressed a wild-type Lm5 and two Lm5 mutants with partially deleted beta3 chains in HEK293 cells. Experiments with the purified Lm5 forms demonstrated that the deletion of the beta3 short arm or its N-terminal domain LN decreases the cell adhesion activity of Lm5, but does not significantly affect the motility activity. A recombinant beta3 short arm protein enhanced integrin-mediated cell adhesion to Lm5 by binding to an unidentified cell receptor. It was also found that the laminin EGF-like domain of the beta3 short arm is a binding site for type VII collagen. These results suggest that the beta3 short arm is involved not only in the matrix assembly of Lm5, but also in its cell adhesion activity. The proteolytic cleavage of the beta3 chain may modulate these functions of Lm5 in vivo. PMID- 16272567 TI - 26 kDa endochitinase from barley seeds: an interaction of the ionizable side chains essential for catalysis. AB - To explore the structure essential for the catalysis in 26 kDa endochitinase from barley seeds, we calculated theoretical pKa values of the ionizable groups based on the crystal structure, and then the roles of ionizable side chains located near the catalytic residue were examined by site-directed mutagenesis. The pKa value calculated for Arg215, which is located at the bottom of the catalytic cleft, is abnormally high (>20.0), indicating that the guanidyl group may interact strongly with nearby charges. No enzymatic activity was found in the Arg215-mutated chitinase (R215A) produced by the Escherichia coli expression system. The transition temperature of thermal unfolding (T(m)) of R215A was lower than that of the wild type protein by about 6.2 degrees C. In the crystal structure, the Arg215 side chain is in close proximity to the Glu203 side chain, whose theoretical pKa value was found to be abnormally low (-2.4), suggesting that these side chains may interact with each other. Mutation of Glu203 to alanine (E203A) completely eliminated the enzymatic activity and impaired the thermal stability (deltaT(m) = 6.4 degrees C) of the enzyme. Substrate binding ability was also affected by the Glu203 mutation. These data clearly demonstrate that the Arg215 side chain interacts with the Glu203 side chain to stabilize the conformation of the catalytic cleft. A similar interaction network was previously found in chitosanase from Streptomyces sp. N174 [Fukamizo et al. (2000) J. Biol. Chem. 275, 25633-25640]; hence, this type of interaction seems to be at least partly conserved in the catalytic cleft of other glycosyl hydrolases. PMID- 16272568 TI - Bacillus circulans MH-K1 chitosanase: amino acid residues responsible for substrate binding. AB - To identify the amino acids responsible for the substrate binding of chitosanase from Bacillus circulans MH-K1 (MH-K1 chitosanase), Tyr148 and Lys218 of the chitosanase were mutated to serine and proline, respectively, and the mutated chitosanases were characterized. The enzymatic activities of Y148S and K218P were found to be 12.5% and 0.16% of the wild type, respectively. When the (GlcN)3 binding ability to the chitosanase was evaluated by fluorescence spectroscopy and thermal unfolding experiments, the binding abilities of both mutant enzymes were markedly reduced as compared with the wild type enzyme. The affinity of the enzyme for the trisaccharide decreased by 1.0 kcal/mol of binding free energy for Y148S, and 3.7 kcal/mol for K218P. The crystal structure of K218P revealed that Pro218 forms a cis-peptide bond and that the state of the flexible loop containing the 218th residue is considerably affected by the mutation. Thus, we conclude that the flexible loop containing Lys218 plays an important role in substrate binding, and that the role of Tyr148 is less critical, but still important, due to a stacking interaction or hydrogen bond. PMID- 16272569 TI - Suppression of multiclade R5 and X4 human immunodeficiency virus type-1 infections by a coreceptor-based anti-HIV strategy. AB - A cyclic chimeric dodecapeptide (cCD) mimicking the conformation-specific domains of CCR5 and CXCR4 was prepared in which Gly-Asp links the amino and carboxyl termini of two combined pentapeptides (S169-G173 of CCR5; E179-R183 of CXCR4) derived from human immunodeficiency virus type-1 (HIV-1) coreceptors. The immunization of Balb/c mice with cCD conjugated with a multiple-antigen peptide (cCD-MAP) induced seven cCD-specific monoclonal antibodies (mAbs, CPMAb-I to VII) that reacted with native CCR5 and CXCR4. Among the tested mAbs, CPMAb-I and II potently inhibited the infection of both the R5 and X4 laboratory strains. CPMAb-III and -VI were effective against only R5 laboratory strains, and also against some X4 and R5 primary isolates. CPMAb-IV and -V had potent antiviral activities against the R5 and X4 primary isolates. In particular, CPMAb-VII was protective against not only R5 and X4 laboratory strains, but also most of the R5 and X4 primary isolates. Moreover, cCD-MAP immunization also induced antibodies that were effective against R5 and X4 multiclade HIV-1 isolates in vitro in two of three cynomolgus monkeys. Taken together, the results suggest that cCD-MAP is a candidate multiclade immunogen that can be used to block multiclade R5 and X4 HIV-1 infections. PMID- 16272570 TI - Solution RNA structures of the HIV-1 dimerization initiation site in the kissing loop and extended-duplex dimers. AB - Dimer formation of HIV-1 genomic RNA through its dimerization initiation site (DIS) is crucial to maintaining infectivity. Two types of dimers, the initially generated kissing-loop dimer and the subsequent product of the extended-duplex dimer, are formed in the stem-bulge-stem region with a loop including a self complementary sequence. NMR chemical shift analysis of a 39mer RNA corresponding to DIS, DIS39, in the kissing-loop and extended-duplex dimers revealed that the three dimensional structures of the stem-bulge-stem region are extremely similar between the two types of dimers. Therefore, we designed two shorter RNA molecules, loop25 and bulge34, corresponding to the loop-stem region and the stem bulge-stem region of DIS39, respectively. Based upon the chemical shift analysis, the conformation of the loop region of loop25 is identical to that of DIS39 for each of the two types of dimers. The conformation of bulge34 was also found to be the same as that of the corresponding region of DIS39. Thus, we determined the solution structures of loop25 in each of the two types of dimers as well as that of bulge34. Finally, the solution structures of DIS39 in the kissing-loop and extended-duplex dimers were determined by combining the parts of the structures. The solution structures of the two types of dimers were similar to each other in general with a difference found only in the A16 residue. The elucidation of the structures of DIS39 is important to understanding the molecular mechanism of the conformational dynamics of viral RNA molecules. PMID- 16272572 TI - High level expression and preparation of autonomous Ca2+/calmodulin-dependent protein kinase II in Escherichia coli. AB - The chymotryptic fragment of Ca2+/calmodulin-dependent protein kinase II (30K CaMKII) is a constitutively active enzyme that phosphorylates a variety of protein substrates in vitro. Although 30K-CaMKII is an often used and powerful tool for protein phosphorylation, the efficient production of catalytically active 30K-CaMKII in Escherichia coli has not yet been successfully realized, probably due to its toxicity in host cells. In this study, we found that a high level expression of 30K-CaMKII as an insoluble form was attained when the N terminal 43 amino acid residues of Xenopus CaMKI were fused to the N-terminal end of 30K-CaMKII (CX-30K-CaMKII). The inactive CX-30K-CaMKII thus expressed in E. coli was reactivated by simple denaturation/renaturation processes and purified on a Ni2+-chelating column. The renatured CX-30K-CaMKII exhibited specific activity similar to that of rat brain CaMKII, and phosphorylated various proteins such as histones, myosin light chain, myelin basic protein, and synapsin I, as in case of 30K-CaMKII or purified CaMKII. Thus, CX-30K-CaMKII, an autonomous CaMKII, can be obtained with a simple procedure using E. coli expression system. PMID- 16272571 TI - Unique molecular architecture of egg case silk protein in a spider, Nephila clavata. AB - We describe a unique silk protein secreted from the cylindrical silk glands of the spider Nephila clavata. This silk is primarily composed of three proteins, whose transcripts of approximately 16.0, 14.5 and 13.0 kb are homologous to one another in two termini and repetitive units, as determined on Northern blotting. Its overall organization shows that it is similar to other characterized silk proteins, including in the mainly central repetitive region as well as the non repetitive N-terminal (166 residues) and C-terminal (176 residues) parts. However, up to 90% of the protein consists of highly ordered repetitive structures that are not found in other silks. The repetitive region mainly consists of several types of complexes and remarkably conserved polypeptide repeats. The assembled repeat units (A1B1) contain a high proportion of Ala (30.41%), Ser (25.15%), and residues with hydrophobic side chains (22.22% for Gly, Leu, Ile, Val and Phe combined). The presence of Ser-rich and GVGAGASA motifs suggests the formation of a beta-sheet. The repetitive region is characterized by alternating arrays of hydrophobic and hydrophilic blocks. The results suggested that this egg case silk is an exceptional protein when compared with previously investigated spider silks. PMID- 16272573 TI - NMR analysis of the Mg2+-binding properties of aequorin, a Ca2+-binding photoprotein. AB - Aequorin, which is a calcium-sensitive photoprotein and a member of the EF-hand superfamily, binds to Mg2+ under physiological conditions, which modulates its light emission. The Mg2+ binding site and its stabilizing influence were examined by NMR spectroscopy. The binding of Mg2+ to aequorin prevented the molecule from aggregating and stabilized it in the monomeric form. To determine the structural differences between Mg2+-bound and free aequorin, we have performed backbone NMR assignments of aequorin in the Mg2+-free state. Mg2+ binding induces conformational changes that are localized in the EF-hand loops. The chemical shift difference data indicated that there are two Mg2+-binding sites, EF-hands I and III. The Mg2+ titration experiment revealed that EF-hand III binds to Mg2+ with higher affinity than EF-hand I, and that only EF-hand III seems to be occupied by Mg2+ under physiological conditions. PMID- 16272574 TI - The role of the cathepsin E propeptide in correct folding, maturation and sorting to the endosome. AB - Cathepsin E (CE) is an endosomal aspartic proteinase of the A1 family that is highly homologous to the lysosomal aspartic proteinase cathepsin D (CD). Newly synthesized CE undergoes several proteolytic processing events to yield mature CE, from which the N-terminal propeptide usually comprising 39 amino acids is removed. To define the role of the propeptide of CE in its biosynthesis and processing, we constructed two fusion proteins using chimeric DNAs encoding the CE propeptide fused to the mature CD tagged with HA at the COOH terminus (termed ED-HA) and encoding the CD propeptide fused to the mature CE (termed DE). Pulse chase analysis revealed that wild-type CE expressed in human embryonic kidney cells is autoproteolytically processed into mature CE within a 12-h chase, whereas the chimeric DE failed to be converted into mature CE even after a 24-h chase. The DE chimera was nevertheless capable of acid-dependent autoactivation in vitro to yield a catalytically active form, although its specificity constants (kcat/Km) were considerably high but less (35%) than those of the wild-type CE. By contrast, the chimeric ED-HA expressed in HeLa cells underwent neither processing into a catalytically active enzyme nor acid-dependent autoactivation in vitro. The ED-HA protein was less stable than wt-CD-HA, as determined on pulse chase analysis and on trypsin digestion. These data indicate that the propeptide of CE is essential for the correct folding, maturation, and targeting of this protein to its final destination. PMID- 16272575 TI - Manipulation of membrane protein topology on the endoplasmic reticulum by a specific ligand in living cells. AB - Almost all integral membrane proteins in the secretory pathway are cotranslationally inserted into the endoplasmic reticulum membrane. Their membrane topology is determined by their amino acid sequences. Here we show that the topology can be manipulated by a factor other than the amino acid sequence. A dihydrofolate reductase (DHFR) domain was fused to the N-terminus of the type I signal-anchor sequence of synaptotagmin II, which mediates translocation of the preceding portion. The DHFR domain was translocated through the membrane in COS7 cells and a transmembrane (TM) topology was achieved. When a DHFR ligand, methotrexate, was added to the culture medium, translocation of the DHFR domain was suppressed and both ends of the signal-anchor sequence remained on the cytoplasmic side. In contrast, translocation of the DHFR domain fused after the signal peptide, which translocates the following region, was not affected by the ligand. The topology-altered fusion protein was anchored to the membrane in a high salt-resistant state, and partially extracted from the membrane under alkali conditions. We concluded that the topology of membrane proteins can be manipulated by a trans-acting factor, even in living cells. PMID- 16272576 TI - WAVE3 functions as a negative regulator of LDOC1. AB - WAVE3 belongs to the Wiskott-Aldrich syndrome protein family that mediates actin reorganization through activation of the Arp2/3 complex. However, the physiological function of WAVE3 is poorly understood. We found that LDOC1-encoded by a gene that is down-regulated in tumor cell lines-binds directly to the verprolin homology domain of WAVE3. Ectopically expressed LDOC1 is localized in the nucleus and induces apoptosis in the cells. This apoptosis is accompanied by an increase in the p53 protein level, but not in p53 transcription, suggesting that LDOC1 inhibits the degradation of p53. Further, the expression of WAVE3 induces the translocation of LDOC1 from the nucleus to the cytoplasm, resulting in the inhibition of LDOC1-induced apoptosis. Thus, it is possible that the LDOC1 function is negatively regulated by WAVE3. PMID- 16272577 TI - Cholinesterase activity of human lung tumours varies according to their histological classification. AB - The probable involvement of acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) in cancer and the relevance of cholinergic responses for lung cancer growth prompted us to study whether cholinesterase activity of human lung is altered by malignancy. Surgical pieces of non-small lung carcinomas (NSLC) and their adjacent non-cancerous tissues (ANCT) were analysed for AChE and BChE activities. AChE activity in adenocarcinoma (AC) was 7.80 +/- 5.59 nmol of substrate hydrolysed per min and per mg of protein (mU/mg), the same as in their ANCT (8.83 +/- 4.72 mU/mg; P = 0.823); in large cell carcinoma (LCC), 7.52 +/- 3.32 mU/mg, approximately 50% less than in their ANCT (15.39 +/- 5.66 mU/mg; P = 0.043); and in squamous cell carcinoma (SCC), 1.39 +/- 0.58 mU/mg, 80% less than in ANCT (6.08 +/- 2.88 mU/mg; P = 0.003). BChE activity was 5.85 +/- 3.20 mU/mg in AC and 9.56 +/- 3.38 mU/mg in ANCT (P = 0.022); 2.94 +/- 2.01 mU/mg in LCC and 6.50 +/- 6.63 mU/mg in ANCT (P = 0.068); and 4.49 +/- 2.30 mU/mg in SCC and ANCT 6.56 +/- 4.09 mU/mg (P = 0.026). Abundant AChE dimers and fewer monomers were identified in lung and, although their distribution was unaffected by cancer, the binding with concanavalin A revealed changes in AChE glycosylation between SCC and their ANCT. The fall in BChE activity affected all molecules, with a strong decrease of the amphiphilic tetramers. Western blotting revealed protein bands with the expected mass of the principal AChE subunits, and the deeper intensity of the protein signal in SCC than in healthy lung, in lanes loaded with the same units of AChE activity, supported an augment in the amount of AChE protein/unit of AChE activity in SCC. The increased availability of acetylcholine in neoplastic lung, resulting from the fall of cholinesterase activity, may enhance cholinergic signalling and contribute to tumour progression. PMID- 16272579 TI - Gallium tris(iodate), Ga(IO3)3. AB - Ga(IO(3))(3) crystallizes in the space group P6(3), with the Ga atom at a site with imposed threefold symmetry. The crystal structure consists of slightly distorted GaO(6) octahedra that are bridged by I atoms of IO(3)(-) groups, giving rise to a three-dimensional polar network. The framework contains unoccupied hexagonal channels running parallel to the hexagonal [001] direction. The iodate groups have their stereochemically active non-bonded electron pairs pointing in the same direction along [001], which creates the polarity in the structure. The I-O bond distances and O-I-O angles are normal, being in the ranges 1.783 (3) 1.847 (2) A and 94.68 (11)-99.61 (12) degrees , respectively. PMID- 16272578 TI - Two catalytic domains are required for protein deacetylation. AB - Histone deacetylase (HDAC)-6 was recently identified as a dual substrate, possibly multisubstrate, deacetylase that can act both on acetylated histone tails and on alpha-tubulin acetylated on Lys40. HDAC-6 is unique among deacetylases in having two hdac domains, and we have used this enzyme as a useful model to dissect the structural requirements for the deacetylation reaction. In this report, we show that both hdac domains are required for the intact deacetylase activity of HDAC-6 in vitro and in vivo. The spatial arrangement of these two domains in HDAC-6 is essential and alteration of the linker region between the two domains severely affects the catalytic activity. Artificial chimeric HDACs, made by replacing the hdac domains in HDAC-6 with corresponding domains from other class II HDACs, show de novo deacetylase activity. Taken together, our results demonstrate for the first time that the spatial arrangement of hdac domains is critical for in vivo deacetylation reaction and may provide a useful model for the development of novel HDAC inhibitors. PMID- 16272580 TI - The K2nCoMo2+nO7+4n family of structures, with n = 2, 3 or 5. AB - The compounds K(4)CoMo(4)O(15) (tetrapotassium cobalt tetramolybdate), K(6)CoMo(5)O(19) (hexapotassium cobalt pentamolybdate) and K(10)CoMo(7)O(27) (decapotassium cobalt heptamolybdate) belong to a series of compounds with closely related crystal structures. In K(4)CoMo(4)O(15), the Co atom and one of the two unique Mo atoms are at sites with threefold symmetry; two of the three unique K atoms lie at sites with 3 symmetry. K(6)CoMo(5)O(19) crystallizes in a new monoclinic structure type. Each [CoO(6)] octahedron is surrounded by one face sharing [MoO(6)] octahedron and six corner-sharing [MoO(4)] tetrahedra. All three compounds have this structural unit in common, but differ in the degree of connectivity between these units. They form layers in K(4)CoMo(4)O(15) and zigzag chains in K(6)CoMo(5)O(19), both by sharing [MoO(4)] tetrahedra. In K(10)CoMo(7)O(27), the structural units are isolated from each other. PMID- 16272581 TI - Diaqua(2,2'-diamino-4,4'-bi-1,3-thiazole)oxosulfatovanadium(IV) tetrahydrate. AB - The title compound, [VO(SO(4))(C(6)H(6)N(4)S(2))(H(2)O)(2)].4H(2)O, displays a distorted octahedral coordination geometry. The 2,2'-diamino-4,4'-bithiazole ligand is present in the usual chelating bidentate mode. The sulfate ligand coordinates in a monodentate fashion to the V atom. A large displacement of the V atom from the equatorial plane towards the oxo group correlates with the strong V=O double bond. In the crystal structure, a three-dimensional supramolecular network is formed by hydrogen bonds. PMID- 16272582 TI - Di-mu-acetato-bis{[2,4-dichloro-6-(2-pyridylmethyliminomethyl)phenolato]zinc(II)} and di-mu-thiocyanato-bis{[2,4-dichloro-6-(2 pyridylmethyliminomethyl)phenolato]copper(II)}. AB - The two title complexes, [Zn(2)(C(13)H(9)Cl(2)N(2)O)(2)(C(2)H(3)O(2))(2)], (I), and [Cu(2)(C(13)H(9)Cl(2)N(2)O)(2)(NCS)(2)], (II), are dinuclear Schiff base compounds. Both molecules are located on crystallographic centres of inversion. In (I), the Zn(II) atom is five-coordinated in a trigonal-bipyramidal coordination, with one imine N atom of one Schiff base and two acetate O atoms defining the basal plane, and one O atom and one pyridine N atom of the Schiff base occupying the axial positions, while in (II), the Cu(II) atom is five coordinated in a square-pyramidal coordination, with one O and two N atoms of one Schiff base and one terminal N atom of a bridging thiocyanate ligand defining the basal plane, and one terminal S atom of another bridging thiocyanate ligand occupying the apical position. The different bridging ligands lead to the different coordinations of the complexes. PMID- 16272583 TI - A novel cadmium(II) coordination polymer with biphenyl-3,3',4,4'-tetracarboxylic acid and 4,4'-bipyridine. AB - A novel cadmium(II) coordination polymer, poly[[[bis(4,4'-bipyridine)cadmium(II)] mu(3)-4,4'-dicarboxybiphenyl-3,3'-dicarboxylato] 0.35-hydrate], {[Cd(C(16)H(8)O(8))(C(10)H(8)N(2))(2)].0.35H(2)O}(n), was obtained by reaction of Cd(CH(3)COO)(2).3H(2)O, 4,4'-bipyridine (4,4'-bpy) and biphenyl-3,3',4,4' tetracarboxylic acid (H(4)L) under hydrothermal conditions. Each Cd(II) atom lies at the centre of a distorted octahedron, coordinated by four O atoms from three H(2)L(2-) ligands and N atoms from two monodentate 4,4'-bpy ligands. Each H(2)L(2 ) ligand coordinates to three Cd(II) atoms through two carboxylate groups, one acting as a bridging bidentate ligand and the other in a chelating bidentate fashion. Two Cd atoms, two H(2)L(2-) anions and four 4,4'-bpy ligands form a ring dimer node, which links into an extended broad zonal one-dimensional chain along the c axis. PMID- 16272584 TI - Why magnesium is five-coordinate in methanol(phthalocyaninato)magnesium(II). AB - The square-pyramidal Mg center in the title compound, [Mg(C(32)H(16)N(8))(CH(4)O)], is five-coordinate due to the formation of back-to back pi-pi dimers that saturate the vacant apical site of the metal coordination sphere. Each complex is a member of a back-to-back and a face-to-face dimer; the latter are tethered by two strong O-H...N hydrogen bonds. The dimers form columns that likely determine the solid-state packing. The phthalocyaninate ligands are essentially planar, with a slight 'hat visor' conformation character. PMID- 16272585 TI - Dichlorooxo(quinoline-8-thiolato-kappa2N,S)(triphenylphosphine oxide kappaO)rhenium(V) acetone solvate. AB - The complex molecule in the title compound, [Re(C(9)H(6)NS)Cl(2)O(C(18)H(15)OP)].C(3)H(6)O, has distorted octahedral geometry. The Re=O bond occupies the position trans to the triphenylphosphine oxide O atom. The Re-Cl bond trans to the thiolate S atom is longer than that trans to the quinoline N atom, implying a stronger trans influence of the S atom. Intra- and intermolecular pi-pi interactions are also observed between the pi rings in the complex. PMID- 16272586 TI - (2,9-dimethyl-1,10-phenanthroline-kappa2N,N')(pyridine-2,6-dicarboxylato kappa3O2,N,O6)copper(II) trihydrate. AB - The copper(II) centre in the mononuclear title complex, [Cu(C(7)H(3)NO(4))(C(14)H(12)N(2))].3H(2)O, is surrounded by one bidentate 2,9 dimethyl-1,10-phenanthroline (dmphen) ligand and one tridentate pyridine-2,6 dicarboxylate ligand, and exhibits a distorted square-pyramidal geometry. The crystal packing involves both hydrogen-bonding and pi-pi interactions. The solvent water molecules link monomers to one another through hydrogen-bonding interactions, forming ladder-like chains in the bc plane. Face-to-face and slipped pi-pi interactions also occur between dmphen rings of neighboring molecules and are responsible for interchain packing. PMID- 16272587 TI - Inclusion of diprotonated [2.2.2]cryptand in the cavity of uranyl-complexed p phenyltetrahomodioxacalix[4]arene. AB - In the title compound, 4,7,13,16,21,24-hexaoxa-1,10 diazoniabicyclo[8.8.8]hexacosane dioxo[7,13,21,27-tetraphenyl-3,17 dioxapentacyclo[23.3.1.1(5,9).1(11,15).1(19,23)]ditriaconta 1(29),5,7,9(30),11(31),12,14,19(32),20,22,25,27-dodecaene-29,30,31,32 tetraolato]uranium dimethyl sulfoxide trisolvate, (C(18)H(38)N(2)O(6))[U(C(54)H(40)O(6))O(2)].3C(2)H(6)OS, the uranyl ion is bound to the four phenoxide groups of the deprotonated p phenyltetrahomodioxacalix[4]arene ligand in a cone conformation, resulting in a dianionic complex. The diprotonated [2.2.2]cryptand counter-ion is located in the cavity defined by the eight aromatic rings of the homooxacalixarene, where it is held by cation-anion, cation-pi and possibly C-H...pi interactions. Dimerization in the packing leads to the formation of sandwich assemblages in which two diprotonated [2.2.2]cryptands are encompassed by two uranyl complexes. PMID- 16272588 TI - Dichlorotetrakis[3-(4-pyridyl)-1H-pyrazole]cobalt(II) acetonitrile tetrasolvate: an infinite hydrogen-bonded network, in an instant. AB - Reaction of 3-(4-pyridyl)pyrazole (4pypz) with cobalt(II) chloride in acetonitrile affords the title complex, [CoCl(2)(C(8)H(7)N(3))(4)].4CH(3)CN, within seconds of addition, as purple X-ray quality crystals. The molecule has C4 symmetry. The metal ion exhibits a trans-N(4)Cl(2) octahedral geometry, with the four 3-(4-pyridyl)-1H-pyrazole ligands coordinating through their pyridyl N-atom donors; one coordinated chloride ion forms hydrogen bonds with the pyrazole rings from four separate units. This configuration creates an infinite three dimensional coordination network containing channels that are filled with acetonitrile solvent molecules. PMID- 16272589 TI - [Di-2-pyridyl ketone N4,N4-(butane-1,4-diyl)thiosemicarbazonato kappa3N,N',S]dioxovanadium(V). AB - The V(V) atom in the title complex, [V(C(16)H(16)N(5)S)O(2)], is five-coordinate in a highly distorted square-pyramidal geometry, with the pyridyl N, the azomethine N and the thiolate S atoms of the di-2-pyridyl ketone N(4),N(4) (butane-1,4-diyl)thiosemicarbazone ligand and one oxo ligand occupying the basal coordination positions, while the second oxo ligand occupies the apical position. The molecules are interconnected by weak intermolecular interactions, mainly of the C-H...O type, involving the oxo atoms. PMID- 16272591 TI - catena-poly[[aqua(3,4,7,8-tetramethyl-1,10-phenanthroline-kappa2N,N')cadmium(II)] mu3-thiosulfato-kappa3S:S:O]. AB - The title compound, [Cd(S(2)O(3))(C(16)H(16)N(2))(H(2)O)](n), presents a polymeric one-dimensional structure running along the P2(1)/c glide direction, with elementary units defined by six-coordinate Cd(II) atoms bonded to three symmetry-related thiosulfate groups, a bidentate tetramethylphenanthroline ligand and one aqua ligand. The bridging thiosulfates bind metal centers through two different sequences, viz. Cd-S-Cd' and Cd'-S'-S'-O'-Cd, defining a closed six membered ring. Individual chains are held together via pi-pi interactions to generate two-dimensional networks parallel to the (100) plane. These, in turn, are connected by much weaker van der Waals interactions. PMID- 16272590 TI - catena-poly[[[diaqua[trans-3-(4-pyridyl)acrylato]samarium(III)]-di-mu-trans-3-(4 pyridyl)acrylato] dihydrate]. AB - In the title compound, {[Sm(4-pya)(3)(H(2)O)(2)].2H(2)O}(n) [4-pya is trans-3-(4 pyridyl)acrylate, C(8)H(6)NO(2)], each Sm(III) atom is ten-coordinated and has a bicapped square-antiprismatic coordination geometry. There is a crystallographic center of symmetry at the mid-point of the Sm...Sm line within each [Sm(4 pya)(3)(H(2)O)(2)](2) dimer. Each dimer is interconnected by two pairs of bridging 4-pya ligands to form a one-dimensional chain. Neighboring chains are connected via hydrogen bonds to form a three-dimensional network. PMID- 16272592 TI - Hydrogen bonding in brucinium dihydrogen citrate trihydrate at 130 K. AB - The structure of brucinium dihydrogen citrate trihydrate (systematic name: 2,3 dimethoxy-10-oxostrychnidinium dihydrogen citrate trihydrate), C(23)H(27)N(2)O(4)(+).C(6)H(7)O(7)(-).3H(2)O, has been determined at 130 K. The crystallographic asymmetric unit comprises two brucinium cations, two dihydrogen citrate anions and six water molecules of solvation. The two citrate anions, which are conformationally dissimilar, associate through extensive hydrogen bonding interactions with the common undulating brucinium cation layer substructures and the water molecules, forming a three-dimensional framework polymer. PMID- 16272593 TI - The hydrogen-bonding network in deacetylcephalothin. AB - The structural analysis of deacetylcephalothin [systematic name: (6R,7R)-3 hydroxymethyl-8-oxo-7-(2-thiophen-2-ylacetylamino)-5-thia-1-azabicyclo[4.2.0]oct 2-ene-2-carboxylic acid], C(14)H(14)N(2)O(5)S(2), shows that the geometry of the central bicyclic moiety is close to the geometry exhibited by other biologically active cephalosporin antibiotics. The molecules are arranged in a helical chain running parallel to the 2(1) axis via a strong O-H...O hydrogen bond. The main helices are zipped together via N-H...O interactions, forming infinite layers. The supramolecular architecture is stabilized by O-H...S and C-H...O hydrogen bonds. PMID- 16272594 TI - O-H...O-bridged dimers linked via C-H...O and C-H...pi interactions in 4,6-di-O benzyl-myo-inositol 1,3,5-orthoformate. AB - The centrosymmetric O-H...O-bonded head-to-head dimers of the title compound, C(21)H(22)O(6), are linked together via bifurcated C-H...O interactions along the a axis and via favourable C-H...pi interactions along the b axis in the crystal structure. PMID- 16272595 TI - 1,4-dimethyl-1,4-diazoniabicyclo[2.2.2]octane diiodide acetonitrile solvate. AB - In the crystal structure of the title compound, C(8)H(18)N(2)(2+).2I(-).CH(3)CN, the dication lies on a mirror plane containing the molecular dication threefold axis. The structure displays C-H...I interactions between H atoms of the 1,4 dimethyl-1,4-diazoniabicyclo[2.2.2]octane dication and the iodide anions. The H...I distances are in the range 2.96-3.18 (4) A. The dications pack forming channels along the b axis, which contain the iodide anions and acetonitrile solvent molecules. PMID- 16272596 TI - Contrasting three-dimensional framework structures in the isomeric pair 2-iodo-N (2-nitrophenyl)benzamide and N-(2-iodophenyl)-2-nitrobenzamide. AB - In 2-iodo-N-(2-nitrophenyl)benzamide, C(13)H(9)IN(2)O(3), the molecules are linked into a three-dimensional framework structure by a combination of a C-H...O hydrogen bond, and iodo-nitro, carbonyl-carbonyl and aromatic pi-pi stacking interactions. In the isomeric compound N-(2-iodophenyl)-2-nitrobenzamide, the framework structure is built from N-H...O, C-H...O and C-H...pi(arene) hydrogen bonds and an iodo-nitro interaction. PMID- 16272597 TI - 2,6-di-tert-butylphenol revisited at 110 K. AB - The title compound, C(14)H(22)O, was studied at 110 K. The phenolic hydroxy group was found to be coplanar with the benzene ring and, due to steric hindrance from the tert-butyl groups, this hydroxy group does not form hydrogen bonds. The shortest intermolecular O...O distance is 3.1008 (11) A, with an O-H...O angle of 117.3 (16) degrees . There are no significant intermolecular pi-pi stacking or C H...pi interactions. PMID- 16272598 TI - 2,2'-(iminodimethylene)bis(1H-benzimidazolium)1+ chloride. AB - The title compound, C(16)H(16)N(5)(+).Cl(-) (nbbH(+).Cl(-)), displays N-H...N, N H...Cl and pi-pi interactions in the crystal packing. The Cl(-) anion is chelated by the nbbH(+) cation via two N-H...Cl hydrogen bonds. Inter-ion N-H...N and N H...Cl hydrogen bonds link ions related by 2(1) screw axes into chains along the c axis. These chains are further linked by glide-plane operations to generate a three-dimensional network, which is additionally stabilized by interchain pi-pi interactions. PMID- 16272599 TI - N-benzylethylammonium nitrate: a three-dimensional hydrogen-bonded framework comprising substructures in zero, one and two dimensions. AB - The title compound is a salt, C(9)H(14)N(+).NO(3)(-), in which two N-H...O hydrogen bonds and two C-H...O hydrogen bonds generate a three-dimensional framework structure. The combination of one N-H...O hydrogen bond and one C-H...O hydrogen bond generates a finite (zero-dimensional) centrosymmetric R(4)(4)(14) aggregate containing two cations and two anions; the combination of the two N H...O hydrogen bonds generates a one-dimensional C(2)(2)(6) chain of alternating cations and anions, and the combination of one N-H...O hydrogen bond and two C H...O hydrogen bonds generates a two-dimensional sheet of alternating R(4)(4)(14) and R(8)(6)(34) rings. PMID- 16272600 TI - Supramolecular structures of three isomeric 4-(methylphenylamino)pyridine-3 sulfonamides. AB - The structures of the three title isomers, namely 4-(2-methylanilino)pyridine-3 sulfonamide, (I), 4-(3-methylanilino)pyridine-3-sulfonamide, (II), and 4-(4 methylanilino)pyridine-3-sulfonamide, (III), all C(12)H(13)N(3)O(2)S, differ in their hydrogen-bonding arrangements. In all three molecules, the conformation of the 4-aminopyridine-3-sulfonamide moiety is conserved by an intramolecular N H...O hydrogen bond and a C-H...O interaction. In the supramolecular structures of all three isomers, similar C(6) chains are formed via intermolecular N-H...N hydrogen bonds. N-H...O hydrogen bonds lead to C(4) chains in (I), and to R(2)(2)(8) centrosymmetric dimers in (II) and (III). In each isomer, the overall effect of all hydrogen bonds is to form layer structures. PMID- 16272601 TI - (RS)-6-ethyl 2-carboxy-1,2,3,4-tetrahydroazulene-6-carboxylate. AB - The title compound, C(14)H(16)O(4), was obtained during the synthesis of 2,6 disubstituted azulene derivatives. In the partially reduced azulene skeleton, the absence of a H atom at the ester substitutent position of the seven-membered ring, as well as lengthened double bonds, indicate a conjugative stabilized system with two overlaid tautomers. PMID- 16272602 TI - N,N',N''-Tris(p-methoxyphenyl)phosphoric triamide. AB - The title compound, C(21)H(24)N(3)O(4)P, is a self-complementary hydrogen-bond (HB) building unit, with (P=)O as the primary HB acceptor and N-H as the HB donor. Each of the four crystallographically distinct and nearly parallel molecules of the unit cell has a net dipole moment along the P=O bond direction and all of the dipoles are directed in the same general crystallographic direction. Head-to-tail N-H...O=P double-HB strands stack adjacent molecules into one-dimensional infinite polar columns. Each polar column is a 2(1) helix and all columns are essentially parallel, resulting in polar order throughout the entire crystal. PMID- 16272603 TI - 4-[2-(4-cyanophenyl)ethenyl]-N-methylpyridinium tetraphenylborate. AB - In the title compound, C(15)H(13)N(2)(+).C(24)H(20)B(-), the pyridyl ring of the cation makes a dihedral angle of 1.6 degrees with the benzene ring. Each is rotated in the same direction with respect to the central -C-CH=CH-C- linkage, by 3.8 and 5.3 degrees, respectively. The anions have a slightly distorted tetrahedral geometry. Molecular packing analysis was carried out using the packing energy portioning scheme in the program OPEC. Around each anion in the crystal structure there are eight anions, which interact with the central anion through C-H...pi interactions. The cations are hydrogen bonded in a head-to-tail fashion, forming chains along [101]. PMID- 16272605 TI - 1:1 complexes of 2,3,5,6-tetrachlorobenzene-1,4-dicarbonitrile with pyrene and phenanthrene: pseudo-isomorphs. AB - 2,3,5,6-Tetrachlorobenzene-1,4-dicarbonitrile forms alternate pi-stacked 1:1 complexes with pyrene, C(8)Cl(4)N(2).C(16)H(10), and phenanthrene, C(8)Cl(4)N(2).C(14)H(10). These complexes are pseudo-isomorphs. Phenanthrene, disordered about a centre of symmetry, takes the position of the pyrene, which sits exactly on this centre of inversion. The tetrachlorobenzenedicarbonitrile molecules in each complex also sit on centres of inversion and are in similar positions within the unit cells in the two structures, except that the orientation of the nitrile groups differs between the two. PMID- 16272604 TI - (Z)-2-(3-methoxybenzylidene)-1-azabicyclo[2.2.2]octan-3-one. AB - The crystal structure of the title compound, C(15)H(17)NO(2), contains two nearly identical but crystallographically independent molecules, each with a double bond connecting an azabicyclic ring system to a 3-methoxybenzylidene moiety. The space group is triclinic P-1. The benzene ring is twisted by 18.44 (5) and 22.35 (4) degrees with respect to the plane of the double bond connected to the azabicyclic ring system for the two molecules. In addition to C-H...pi interactions, molecules are held together in the solid state by van der Waals interactions. PMID- 16272606 TI - 6-phenyl-3-(4-pyridyl)-1,2,4-triazolo[3,4-b][1,3,4]thiadiazole. AB - The title compound, C(14)H(9)N(5)S, has been synthesized and characterized both spectroscopically and structurally. The triazolo-thiadiazole system, the pyridine ring and the phenyl ring are all planar. The plane of the triazolo-thiadiazole system forms dihedral angles of 1.53 (13) and 7.55 (12) degrees with the planes of the pyridine and phenyl rings, respectively. In the molecule, there are two intramolecular interactions of types C-H...N and C-H...S. Intermolecular C-H...N interactions involving a phenyl CH group and a triazole N atom lead to the formation of a one-dimensional chain. In the crystal structure, two types of pi pi interactions affect the packing of the molecules. In addition, there are intermolecular non-bonded S...N contacts of 2.870 (2) A, which may cause steric hindrance. PMID- 16272607 TI - Two phosphate-imidazole complexes with and without a hydrogen-bonded guest molecule. AB - The molecular structures of the complexes imidazolium 6,6'-di-tert-butyl-4,4' dimethyl-2,2'-thiodiphenyl phosphate, C(3)H(5)N(2)(+).C(22)H(28)O(4)PS(-), (I), and imidazolium 6,6'-di-tert-butyl-4,4'-dimethyl-2,2'-thiodiphenyl phosphate diisopropyl hydrazodicarboxylate hemisolvate, C(3)H(5)N(2)(+).C(22)H(28)O(4)PS( ).0.5C(8)H(16)N(2)O(4), (II), have been determined. While (I) forms the expected hydrogen-bonded chain utilizing the two imidazole N-bound H atoms, in (II), the substituted hydrazine solvent molecule inserts itself between the chains. Compound (I) exhibits a strong N-H...O hydrogen bond, with an N...O distance of 2.603 (2) A. The hydrazine solvent molecule in (II) lies about a twofold axis and the N-bound H atoms are involved in bifurcated hydrogen bonds with phosphate O atoms. A C-bound H atom of the imidazolium cation is involved in a C-H...O interaction with a carbonyl O atom of the hydrazine solvent molecule. PMID- 16272608 TI - Magnesium in drinking water and cardio-vascular disease--an epidemiological dilemma. AB - The relation between drinking water and cardiovascular disease has been evaluated in a number of studies. Certain of these show strong relationships whereas others do not. This review analyses the methodological aspects on epidemiological studies in terms of dose-range, confounding factors and multiple exposures. It is concluded that there is good evidence for a relation between drinking water quality and cardio-vascular disease, that several methodological criteria for such studies need to be considered, that there is little evidence for magnesium as the single causative agent, and that the relevant exposure should be considered as a mixture of minerals. PMID- 16272609 TI - [Magnesium for nutrition]. AB - Magnesium plays an essential role in a very wide range of fundamental cellular reactions. The deficiency in the organism may lead to serious biochemical and symptomatic changes. In 1932, McCollum and coworkers made the first systematic observations of magnesium deficiency in rats and dogs. In 1934, the first description of clinical depletion in man was published in patients with various underlying diseases. In the early 1950, Flink and coworkers documented depletion of magnesium ion in alcoholics and in patients on magnesium-free i.v. solutions. In this paper, the biochemical and nutritional function, and nutritional essentiality for magnesium are reviewed. PMID- 16272610 TI - [Homeostasis and magnesium--comparison with calcium]. AB - Although Mg and Ca, bivalent cations commonly coexisting in nature and living organism, share some similar chemical properties, the role played by Mg in biological regulation and signal transduction is far less important than that by Ca. In contrast to the vast extra- and intra-cellular concentration gradient of Ca as high as 10,000, intracellular Mg is only 3 times as high as extracellular Mg. Systemic Mg metabolism including serum Mg only controlled by renal tubular reabsorption is not as precisely regulated as the Ca counterpart most precisely controlled by parathyroid hormone (PTH), 1,25 (OH)(2) vitamin D and bone. Such difference between Mg and Ca is probably based on the properties of these two kinds of atoms. Mg has a smaller ionic radius with electron arrangement making protein binding more difficult and velocity constant for water binding smaller. Signal transduction by Mg is thus destined to be much less efficient than that by Ca. PMID- 16272611 TI - [Aging and magnesium]. AB - The aim of this article is to clarify the relationships between aging or age related diseases and magnesium (Mg). The mutation of mitochondrial DNA can occur in both aging and Mg deficiency, resulting in peroxidation, intracellular Ca(2+) accumulation and apoptosis. The capability against peroxidation decreases in aging and Mg deficiency. DNA polymerase I, RNA polimerase and DNA helicase require Mg for their activities. Under Mg deficiency the replication, transcription and translation of DNA become erroneous. The dysfunctions of vascular endothelial cell occur in aging and Mg deficiency. In comparison between adult and old rats using the rings of rat thoracic aortae, vasorelaxation by acetylcholine and isoproterenol is lower in old rats, but can be improved through high Mg concentration. Although women with menopause are suffered from osteoporosis due to estrogen deficiency, bone fragility increases with additive Mg deficiency. High Ca intake is recommended for women with menopause, but adequate Mg intake is necessary to lower dietary Ca/Mg ratio, because the high ratio prompts blood coagulation. About lipid metabolism Mg can play a statin-like activity. Mg deficiency is complicated with lifestyle-related diseases, osteoporosis, bone fragility, depression and elderly dementia. PMID- 16272612 TI - [Growth and magnesium]. AB - Magnesium is one of essential minerals for humans and animals. Mg has important roles in energy metabolism, protein synthesis, bone metabolism and muscle contraction. Mg deficiency causes tetany, muscular incoordination, growth retardation and death. Mg is necessary for normal growth, however the mechanism is unclear. In this article, we will describe Mg metabolism, Mg deficiency, and the relationship Mg and growth. PMID- 16272613 TI - [The calcium receptor and magnesium metabolism]. AB - The calcium receptor (CaR), which regulates Ca(2+) homeostasis, senses Mg(2+) and participates in Mg(2+) metabolism mainly at the parathyroid gland and kidney as well as Ca(2+) metabolism. In the parathyroid gland, Mg(2+) suppresses parathyroid hormone (PTH) secretion by the activation of CaR, and regulates the sensitivity of CaR by its unique effect. In the kidney, the activation of CaR inhibits Mg(2+) reabsorption directly or indirectly, mainly at the downstream of loop of Henle. PMID- 16272614 TI - [Nuclear Receptor PPARs and magnesium]. AB - Peroxisome proliferator-activated receptors (PPARs) are nuclear hormone receptors that are activated by fatty acids and their derivatives. PPARs consist of three isotypes named PPAR alpha (NR1C1), PPAR beta/delta (NR1C2) and PPAR gamma (NR1C3) in vertebrates. Each of them is encoded in a separate gene and binds fatty acids and eicosanoids. Although each isotype fulfills distinct functions, PPARs function not only as an important fatty acid sensor that regulate lipid, carbohydrate and amino acid metabolism but also play an important role in various signaling pathways (immunity, inflammation, apoptosis and cell differentiation). Dysfunction of PPAR-mediated signals leads to various diseases such as diabetes, obese, hyperlipidemia, inflammation and cancer. Importantly, magnesium appears to play a pivotal role in regulating the PPAR-mediated signaling pathways as a key cofactor in the protein phosphorylation. Therefore, restrict control of magnesium concentration in the body appears to be very important for protection for these diseases. In this review, I focus on emerging knowledge about relationship between PPAR-mediated signals and magnesium. PMID- 16272615 TI - [Lipid metabolism and magnesium]. AB - According to the Vital Statistics Report published by the Japanese Ministry of Health and Welfare, heart disease and cerebrovascular disease are the main causes of death in Japan. The main pathological finding in these diseases is atherosclerosis and the main risk factors, besides the patient's age and diathesis, include hyperlipidemia, hypertension, diabetes, obesity and smoking. Among the aforementioned various risk factors, hyperlipidemia play a crucial role at the stage of atherosclerosis. The main pathological findings in atherosclerosis include abnormal reactions of neutrophils, lymphocytes and monocytes/macrophages, vascular smooth muscle cells and vascular endothelial cells, and the accumulation of cholesterol ester in the arterial wall. Previously, Mg(2+) deficit and the lower blood concentration of Mg(2+) was a frequent in patients with the main risk factors, hyperlipidemia, hypertension, diabetes, and obesity. Magnesium is necessary the activity of lecithin cholesterol acyltransferase (LCAT) and lipoprotein lipase (LPL), which lowers triglyceride levels and raises HDL-cholesterol levels. Moreover, Mg(2+)-ATP is also the controlling factor for the rate-limiting enzyme in the cholesterol biosynthesis, which associated with cholesterol levels. In this article, we first discuss the effect of Mg(2+) deficit on atherosclerosis, especially hyperlipidemia in bloodstream and liver. Then, based on recent studies including our own, we describe the Mg(2+) deficit and the relationships between risk factors for atherosclerosis, hypertension, oxidative stress, cholesterol reverse transport system, and the molecular mechanisms, especially peroxisome preoliferator-activated receptor (PPAR), which have the pleiotropic effect in atherosclerosis. The mechanism is likely the effect of Mg(2+) on atherosclerosis. PMID- 16272617 TI - [Recent Topics for magnesium mobilization in neurons]. AB - Although magnesium ion (Mg(2+)) is believed as an important factor for maintenance of normal neuron function, our understanding of its intracellular mobilization and physiological function is still not enough. We need to know the following mobilization mechanisms of Mg(2+) in neurons:Mg(2+) influx via ion channels, Mg(2+) release from intracellular pools, and Mg(2+) transporters located on cell membrane. In this short review, I show the recent findings of intracellular Mg(2+) release and also Na/Mg antiport on cell membrane in PC12 cells that have been revealed by fluorescent Mg imaging techniques. PMID- 16272616 TI - [Role of magnesium in cardiac metabolism]. AB - Numerous studies have suggested that magnesium ion (Mg(2+)) plays an important role in pathogenesis of cardiovascular diseases including hypertension, ischemic heart disease. Hypomagnesemia is often associated with the imbalance of sodium, potassium, and calcium ions. Magnesium deficiency can occur frequently in alcoholics, and in patients with hypertension, congestive heart failure, myocardial infarction. Magnesium deficiency induces an increase in intracellular Ca(2+) concentration in cardiac myocytes, formation of reactive oxygen species, production of inflammatory cytokines, leading to the development of ischemic heart disease, congestive heart disease, sudden cardiac death, atherosclerosis, and arrhythmia. In addition, catecholamines can evoke marked Mg(2+)efflux which is associated with a concomitant increase in the force of contraction of the heart. While many of the mechanisms remains elusive, the beneficial effects of magnesium on the myocardium appear to be convincing. Further studies will be necessary to elucidate the molecular basis of the cardio-protective effects of magnesium. PMID- 16272618 TI - [Bone and magnesium]. AB - The half of magnesium exists in the bone. Although the long-term magnesium deficiency causes osteoporosis, most of them are accompanied by complications such as malabsorption syndrome or alcohol abuse. The animal models of magnesium deficiency show that the trabecular number or thickness is reduced, and the histological evaluation revealed that the bone formation is reduced, and bone resorption is enhanced. The trabecular bone volume, maximum load and elastic modulus are reduced. Degree of mineralization and trabecular microhardness are decreased in the hemo-dialysis patients, suggesting that bone quality is exacerbated. The magnesium takes part in the various chemical reactions in the body, but its mechanism of the regulation and the effect on the metabolism of bone are not completely understood. PMID- 16272619 TI - [Metabolic syndrome and magnesium]. AB - With westernization of lifestyle in Japanese people, dietary intake of Mg by grain, barley, seaweed, vegetable, and nuts has been remarkably diminished. Resultantly, Japanese people might develop hypomagnesemia easily. Likewise, upon drastic change of Japanese lifestyle, metabolic syndrome has been increasing a bigger problem of Japanese health in recent days, probably resulting from various causes, such as increasing intake of animal fat, exercise insufficiency, and accumulation of various stresses. People with metabolic syndrome are often complicated with obesity, hypertension, hyperglycemia, and hyperlipidemia, and thus be susceptible to cardiovascular events. Hypomagnesemia may cause an increase of vascular tonus by intracellular magnesium depletion, resulting in an increase of blood pressure. Furthermore, it might cause impaired insulin secretion, insulin resistance, and hyperlipidemia, and finally leading to the development of metabolic syndrome. Therefore, the importance of magnesium intake for the maintenance of health should be increasingly recognized. PMID- 16272620 TI - [Fetus and magnesium]. AB - Chronic magnesium deficiency in pregnant women is frequently seen because of inadequate or low intake of magnesium. Magnesium deficiency during pregnancy can induce not only maternal and fetal nutritional problem, but also pediatric consequences that might last throughout life. Many epidemiological studies have disclosed that restricted fetal growth, i.e., intrauterine growth retardation (IUGR) is associated with an increased risk of insulin resistance in adult life. We previously postulated that intracellular magnesium of cord blood platelets is lower in the small for gestational age than in the appropriate for gestational age group, suggesting chronic intrauterine magnesium deficiency may result in IUGR. Taken together, chronic intrauterine magnesium deficiency in the fetus may lead to or program the insulin resistance after birth. Prospective study whether the children born with magnesium induced IUGR are at high-risk for metabolic syndrome in childhood or adulthood is currently undertaken. PMID- 16272621 TI - Magnesium and myocardial infarction. AB - The data on magnesium supplementation in patients with acute myocardial infarction (AMI) is conflicting. Although a number of relatively small randomized clinical trials have demonstrated a remarkable reduction in mortality when administered to relatively high risk AMI patients, two recently published large scale randomized clinical trials (the Fourth International Study of Infarct Survival and Magnesium in Coronaries) failed to show any superiority of intravenous magnesium over placebo. Nevertheless, the theoretical potential benefits of magnesium supplementation as a cardioprotective agent in coronary artery disease (CAD) patients, in conjunction with previous promising results from work in animal and humans, its relatively low cost, easy administration, with no need for special expertise, and relatively free of adverse effects, gives magnesium a place in treating CAD patients, especially high-risk groups such as CAD patients with heart failure, the elderly and hospitalized patients with hypomagnesemia. PMID- 16272622 TI - [Role of magnesium in hypertension therapy]. AB - Magnesium is the second most abundant intracellular cation, and the important element that has numerous biological functions in the cardiovascular system. Furthermore, magnesium acts as calcium antagonist, regulates the calcium metabolism, and finally participates in blood pressure control. Although the evidence linking magnesium intake to the prevention of cardiovascular diseases was gathered, data from clinical studies about the magnesium usage in hypertension therapy have not been conclusive. The present review discusses the novel molecular and pharmacological effects of magnesium on cardiovascular systems and the implications in the clinical hypertension therapy based on the evaluating of patients' magnesium status. PMID- 16272623 TI - Potassium and magnesium depletions in congestive heart failure--pathophysiology, consequences and replenishment. AB - Congestive heart failure (CHF) is becoming more frequent worldwide. Both potassium (K) and magnesium (Mg) deficiencies are common and can be associated with risk factors and complications of heart failure (HF). The major causes of K and Mg depletions are the effects of compensatory neuroendocrine mechanisms (activation of the renin-angiotensin-aldosterone and sympathoadrenergic systems), digoxin therapy, and administration of thiazide or loop diuretic therapy in CHF. Particular attention should be paid to K and Mg restoration in CHF, because of the consequences of both deficiencies (increased arrhythmic risk, vasoconstriction), and the co-supplementation of both ions is necessary in order to achieve K repletion. Mg and K should be employed as first-line therapy in digitalis intoxication and drug-related arrhythmias, and should be considered an important adjuvant therapy in diuretic treated patients with CHF. Another possibility to restore normal K and Mg status is usage of a K, Mg sparing diuretics. PMID- 16272624 TI - [Treatment of adynamic bone disease with the complete replacement from calcium carbonate to sevelamer hydrochloride]. AB - The aim of this study was to examine the therapeutic effect of hypocalcemic stimulation caused by sevelamer hydrochloride (SH) administration on adynamic bone disease (ABD). The subjects were 28 maintenance hemodialysis (HD) patients who had remained in ABD state in spite of no administration of vitamin D(3) since HD induction (15 males and 13 females;12 diabetic patients and 16 non-diabetic patients). The mean age was 61.8+/-9.5 years and the mean HD duration was 5.5+/ 3.9 years. The calcium concentration in the dialysate was 3.0 mEq/L. We made the final daily dose of SH after two months the same as the first daily dose of calcium carbonate (CC) in the following manner. At first we administered only CC at breakfast and lunch and SH at supper. And for the next two weeks we administered CC at breakfast and SH at lunch and supper. And for the final two weeks we administered only SH. After that we increased the dose of SH as much as possible. We evaluated the therapeutic effect of the above treatment on ABD using intact-osteocalcin (iOC) [Teijin. Tokyo] as a marker before and 6, 12 months after the beginning of the replacement. If iOC Ievel of 30 to 70 ng/mL showed normal tumover bone (NTB), 5 cases (17.9%) changed into NTB in 6 months. 9 cases (32.1%) changed into NTB in 12 months and one case (3.6%) changed into ostitis fibrosa in 12 months. It is thought that SH is effective for the treatment of ABD but we have to be careful for ostitis fibrosa. PMID- 16272625 TI - [Treatment with phosphate binder (sevelamer hydrochloride, calcium carbonate) based on PTH]. AB - When sevelamer hydrochloride is used as a phosphate binder instead of the more common calcium carbonate, the PTH rises. This has been observed in many cases and makes it more difficult for practical use. However, considering the calcium load, the excessive dosing of calcium carbonate must be avoided. With that in mind, we divided the i-PTH in groups of 100 pg/mL and tested the changes in i-PTH value, P value, the adjusted Ca value, and the product of Ca and P before and after a dosage of sevelamer hydrochloride. When the average i-PTH was under 100mg/mL 6 months before dosing, the sevelamer hydrochloride single dosage group showed an early rise in i-PTH after dosing, maintaining a higher level than the calcium carbonate combined dosage group. Therefore, it was concluded that the use of sevelamer hydrochloride alone as a phosphate binder is best. On the other hand, the group with an average i-PTH over 100 pg/mL 6 months before dosing showed a rise of i-PTH that went over the K/DOQI guideline with a single dosing of sevelamer hydrochloride. Therefore, we concluded that using both sevelamer hydrochloride and calcium carbonate for phosphate binder is best. PMID- 16272626 TI - [Defining the role of sevelamer chloride as a therapeutic agent for management of phosphate in patient with hemodialysis]. AB - It has been about 18 months after the commercial release of sevelamer hydrochloride. I examined the phosphorus management in patient with hemodialysis with the use sevelamer hydrochloride. The phosphorus reduction of sevelamer hydrochloride was weaker than the precipitated calcium carbonate. Patients on artificial dialysis must undergo dietary therapy for the sevelamer hydrochloride to be effective. However, by switching from precipitated calcium carbonate to sevelamer hydrochloride, the calcium level dropped, and as a result the intact PTH rose. It was possible to subdue the rise of intact PTH with vitamin D. The usefulness of vitamin D dramatically increased with the decrease and the eventual cancellation of precipitated calcium carbonate. Furthermore, the calcium load lightened and the product of calcium and phosphorus decreased. At this point, I do not expect substantial phosphorus reduction from sevelamer hydrochloride. It must be recognized that the basis of phosphorus management is proper diet. Only with that can sevelamer hydrochloride be used to its fullest potential. PMID- 16272628 TI - [Fibroblast growth factor (FGF)-23 in patients with hypoparathyroidism]. AB - Hypoparathyroidism is a well-described cause of hyperphosphatemia. We aimed to clarify the physiological role of FGF-23 in serum phosphate homeostasis in hypoparathyroidism after thyroidectomy. Increased serum FGF-23 levels were found in patients with hyperphosphatemia and hypocalcemia, caused by hypoparathyroidism after thyroidectomy. After the recovery of parathyroid function, the serum level of calcium, phosphate, and FGF-23 was normalized. Serum FGF-23 levels were significantly higher in patients with permanent hypoparathyroidism than in healthy controls. These results indicate that FGF-23 may play an important role in serum phosphate homeostasis by its up-regulation in the hyperphosphatemic condition. PMID- 16272627 TI - [Effects of intermittent treatment with sevelamer hydrochloride on parathyroid hyperplasia and vascular calcification in rats with chronic kidney disease]. AB - Phosphorus directly controls parathyroid hormone (PTH) synthesis and secretion. Serum levels of the novel phosphate-regulating hormone, fibroblast growth factor 23 (FGF23), are positively correlated with hyperphosphatemia in patients with chronic kidney disease (CKD). Rats were fed a diet containing adenine for 4 weeks to establish CKD. Animals were then offered a diet containing sevelamer hydrochloride (sevelamer) or a normal diet for alternating 2 week periods over 8 weeks. Adenine-treated rats showed marked elevations of serum phosphorus, PTH and FGF23 levels associated with parathyroid hyperplasia and aortic calcification. Serum phosphorus, PTH and FGF23 levels decreased rapidly when sevelamer treatments commenced and recovered rapidly once they were discontinued. However, intermittent treatment with sevelamer successfully inhibited parathyroid hyperplasia and aortic calcification. In conclusion, phosphate-binder treatment can effectively inhibit the elevation of serum FGF23 levels, as well as PTH levels, under conditions of CKD. Setting up a period of reduced serum phosphorus levels, even if it is intermittent, is worthwhile for the inhibition of the development of parathyroid hyperplasia and aortic calcification. PMID- 16272629 TI - [Usefulness of power Doppler ultrasonography for the diagnosis of autoinfarction of parathyroid gland in secondary hyperparathyroidism]. AB - Spontaneous remission due to parathyroid infarction of secondary hyperparathyroidism is rare compared with that of primary hyperparathyroidism, probably because several glands are enlarged in secondary hyperparathyroidism. Lately, neck ultrasound examination has become a more beneficial and specific method for the diagnosis of enlarged parathyroid glands in contrast to classic diagnostic techniques such as computed tomography (CT), magnetic resonance imaging (MRI) and scintigraphy. However, the diagnosis of parathyroid infarction reported in previous studies was often based on CT, MRI and scintigraphy findings and there are few studies that reported such diagnosis by urgent power Doppler ultrasonography of the neck. Here we present a hemodialysis patient with autoinfarction of the left parathyroid gland diagnosed by urgent power Doppler ultrasonography of the neck. PMID- 16272630 TI - [Two patients with hyperparathyroidism which was progressive for more than ten years after successful kidney transplantation]. AB - It is well known that in some patients advanced renal hyperparathyroidism (HPT) persists after successful kidney transplantation (RTx) and in those patients parathyroidectomy is required usually within one year after RTx. We experienced two patients with advanced renal HPT which released after successful RTx and parathyroidectomy was performed in more than 10 years after RTx, even their kidney function was well-preserved. Hypercalcemia was gradually progressive for ten years after RTx and enlarged parathyroid glands were detected by image diagnosis. We performed parathyroidectomy and HPT was dramatically improved. It is possible that primary HPT occurred de novo after RTx or renal HPT was progressive. We evaluated those possibility based on histopathological findings dramatically improved of removed parathyroid glands. PMID- 16272631 TI - [Apoptosis: a possible mechanism of suppressing parathyroid hyperplasia by calcimimetics]. AB - The mechanism through which calcimimetic compounds suppress parathyroid cell growth has not been fully elucidated. We investigated the effect of the calcimimetic compound (NPS R-568:R568) on the parathyroid cell growth in vitro. Whole parathyroid glands of subtotally nephrectomized rats fed high phosphorus diet for 8 weeks were used in this study. Fresh rat parathyroid glands were incubated in a media (phosphorus concentration is 1.0 mM and calcium is 1.25 mM) with R-568 (10-4 M) or calcitriol (10-7 M) or vehicle for 6 hours. Medium PTH level was significantly decreased in both R568-treated group and calcitriol treated group compared with either vehicle-treated group, or no-treated group. While TUNEL-positive cells were similar in calcitriol-treated group compared with both vehicle-treated and no-treated groups, they were significantly increased in R568-treated group compared with other three groups. These results suggest that calcimimetic compounds, NPS R-568 reduces PTH secretion and induces apoptosis of hyperfunctional parathyroid cell in vitro. PMID- 16272632 TI - [Correlation of serum Bio-intact PTH (1-84) and parathyroid gland size in hemodialysed patients]. AB - Bio-intact parathyroid hormone (Bio-PTH) assay, which measures exclusively intact PTH (1-84) molecule, provides a better assay for estimating parathyroid function in hemodialysis (HD) patients, whereas intact PTH (I-PTH) assay cross-react with PTH (7-84) as well as PTH (1-84). We have found that PTH (7-84) accumulated into serum of hemodialysis patients due probably to its impaired excretion into urine. We have reported that parathyroid gland size is one of major predictor for vitamin D responsiveness in secondary hyperparathyroidism. Therefore, we investigated whether serum Bio-PTH, in comparison with serum I-PTH, may provide a relevant assay to estimate parathyroid function as evidence by its correlation with parathyroid gland size on ultrasound examination. PMID- 16272633 TI - [High prevalence of secondary hyperparathyroidism due to vitamin D insufficiency in Graves' disease]. AB - We studied concentrations of serum calcium, intact parathyroid hormone (iPTH), and 25 hydroxyvitamin D (25 [OH] D) in 343 patients with Graves' disease (273 women, 70 men) during one year period. The serum 25 (OH) D levels were 37+/-19 nmol/L and vitamin D deficiency (defined as a serum 25 (OH) D value below 25 nmol/L) was found in 35% of the patients. There was a significant seasonal variation in the 25 (OH) D concentrations, with high values of 51+/-21 nmol/L during the summer term (July-September), and with low values of 26+/-17 nmol/L during the winter term (January-March). The average serum iPTH level was 48+/-26 pg/mL, with 26% (89/343) having values above the normal range. Our results showed high prevalence of secondary hyperparathyroidism in patients with Graves' disease during antithyroid drug therapy and vitamin D and/or calcium supplements are recommended for patients with vitamin D deficiency. PMID- 16272634 TI - [Case of autosomal dominant polycystic kidney disease associated with pelvic insufficiency fracture]. AB - We experienced a case of A-59-year-old woman having autosomal dominant polycystic kidney disease with renal insufficiency associated with pelvic insufficiency fracture. On admission the pelvic bone compressed by the enlarged kidney and liver due to polycystic disease was demonstrated on the pelvic CT. Her bone mineral density was not significantly decreased contrary to our expectation. The prominent bone absorption in the endosteal and exosteal surface of the cortical bone was demonstrated in the bone specimen form iliac bone although the state of the cancellous bone and bone formation rate was normal. We performed the transcatheter embolization (TAE) to the kidney and liver to diminish their sizes. The fracture was rapidly improved and she could walk 6 months later after the TAE. In this case, we considered that the longstanding compression to the pelvic bone by the enlarged liver and kidney made the cortical and the binding of muscle/tendon and cortical bone fragile and it led to the pelvic insufficiency fracture. PMID- 16272635 TI - [Risedronate: a possible treatment for extraosseous calcification]. AB - A case report of 51 year-old female, diagnosed as CREST syndrome, presenting with an ectopic calcification in the left shoulder joint, which disappeared soon after the start of risedronate. She had been taking steroid and NSAIDs for the past four years, but the pain and the range of motion of her shoulder became worse and restricted progressively during the last three years only to form extraosseous calcification. Laboratory data showed normal renal function, no inflammatory changes, and no abnormalities in calcium and and phosphate metabolism including parathyroid hormone. Risedronate was administered for glucocorticoid-induced osteoporosis. Although the bone turnover markers, such as serum NTX (N-terminal telopeptides of type I collagen) and BSAP (bone specific alkaline phosphate), did not show remarkable changes, the pain disappeared a week later and the range of motion recovered a month later. The X-ray at 6 months risedronate treatment revealed a complete disappearance of the ectopic calcification. Risedronate, probably through a different process from etidronate, could prevent extraosseous calcification. PMID- 16272636 TI - [Suspected case of pseudohypoparathyroidism type II]. AB - We experienced a suspected case of pseudohypoparathyroidism type II. The patient came to our emergency room with no thermal convulsion. The Ellsworth-Howard test was applied to the patient to determine the type of PHP. PMID- 16272637 TI - [Recurrence of sarcoidosis in a hemodialysis patient confirmed by abnormal calcium metabolism]. AB - 28-year-old man hospitalized for a fever of unknown origin. This patient was already diagnosed as neurosarcoidosis proven by brain biopsy in 1997, then entered chronic hemodialysis therapy in 2002. The data showed hypercalcemia without taking any calcium agent and vitamin D, also showed suppressed intact parathyroid hormone and normalized 1,25-dihydroxyvitamin D even the condition of end stage renal failure. Recurrence of sarcoidosis was made in this hospitalization. The laboratory data as well as symptom improved after taking oral prednisolone. We reported the case of recurrent sarcoidosis in a hemodialysis patient confirmed by abnormal calcium metabolism. PMID- 16272638 TI - [2 dialysis cases which came to our clinic for the symptoms caused by hypercalcemia]. AB - We'll report 2 dialysis cases which came to our clinic for the symptoms caused by hypercalcemia. Patients complained of sleeplessness, itching, headache, palpitation, apathy, akinesis, leanness, foot gangrene and so on. Hypercalcemia is one of the complication of vitamin D and calcium carbonate administration in chronic renal failure, though the frequency and risk are not clearly documented. Hypercalcemia aggravates the outcome of patients on dialysis and contributes to vascular calcification in long term. Recently various factors involving cardiovascular calcification are discussed, but first of all we must be very careful for the symptoms of hypercalcemia, and careful monitoring of plasma calcium concentration are recommended. PMID- 16272639 TI - [Effects of raloxifene on bone biomarkers in postmenopausal women on maintenance haemodialysis]. AB - Postmenopausal women on maintenance haemodialysis (MHD) has considerably higher risk of bone fracture than general population with combination of postmenopausal osteoporosis and renal osteodystrophy. However, the treatment of osteoporosis on MHD has not been established. Evidence indicates raloxifene (RLX), a selective estrogen receptor modulator, is effective for a protection of bone fracture without increasing of breast cancer and endometrial cancer. We hereby report short-term use experience of RLX for the postmenopausal MHD patients. Fifteen postmenopausal MHD patients with less than 80% of YAM bone density in DEXA administrated 60 mg RLX on every HD days (3 days/week). Serum NTX level significantly decreased after 6 months (180 +/- 18 vs. 95 +/- 12 nmol/BCE/L, p< 0.05), however, i-PTH did not have the significant difference. (115 +/- 23 vs. 157 +/- 29 pg/mL). RLX is effective for bone biomarker improvement in postmenopausal MHD patients. Further evaluation for the effectiveness and safety of RLX is necessary in the long term. PMID- 16272640 TI - [Effects of parathyroid hormone gene polymorphism on cardiovascular mortality]. AB - It is well-known that secondary hyperparathyroidism of uremia influences not only bone and mineral metabolism but also cardiovascular complications. Here we reported the effects of the level of serum intact PTH and its gene polymorphism on cardiovascular and non-cardiovascular mortality in hemodialysis patients. We analyzed the association between clinico-molecular parameters and 3-year survival in 508 hemodialysis patients among whom 90 patients died. The multivariate Cox proportional hazards models showed that the presence of diabetes mellitus, levels of albumin and intact PTH, and BstB I genotype were indicated as independent predictors of cardiovascular mortality, whereas age and albumin level were indicated as those of non-cardiovascular mortality, suggesting that the level of intact PTH and its gene polymorphism effect cardiovascular mortality in hemodialysis patients. PMID- 16272641 TI - [Influence of serum phosphate level for effects of OCT pulse therapy in hyper-PTH HD patients]. AB - When we treat hyper-PTH patients with OCT pulse therapy, it is easy to suppress the PTH secretion below 30 ng/mL by controlling phosphate concentration under 5 mg/dL. But in hyperphosphate (>5.0 mg/dL) cases, we can't stop or avoid the secretion of PTH by OCT pulse therapy. PMID- 16272642 TI - [Clinical investigation of the effect of blood sampling day on serum phosphorus concentration]. AB - Whether the presence or absence of notification about the blood sampling date affects the serum phosphate concentration was examined. After periodical sampling at the beginning of the week with 2-day interval for regular dialysis, the blood was sampled on the midweek day with 1-day interval for dialysis upon the patient's consent right before sampling without an advance notification. In the next month subsequently periodical sampling at the beginning of the week and the sampling on the midweek day were performed after notification of the twice blood sampling before 1 week in advance. Then both serum phosphorus data were comparably investigated. In case the sampling was not notified, there is no difference in the serum concentration between the beginning of the week and the midweek day. On the other hand, in case the sampling was notified, the serum concentration significantly decreased on the weekday compared to that at the beginning of the week. It therefore was risky that the serum phosphorus concentration at the beginning of the week with 2-day interval for dialysis was presumed to be higher value than that on the midweek day with 1-day interval for dialysis. We need to consider the possibility that the presence or absence of notification about the blood sampling date affects the level of serum phosphorus concentration. PMID- 16272644 TI - [Clinical analysis of time-course changes in minerals and parathyroid hormone levels during treatment with phosphate binders in hemodialysis patients with secondary hyperparathyroidism]. AB - 146 hemodialysis (HD) patients with secondary hyperparathyroidism (2 degrees HPT) were studied about the therapeutic effects of phosphate binders. We divided these patients into four groups; Group I: 59 patients treated with CaCO(3) (1.5-6.0 g/day), Group II: 42 cases with sevelamer hydrochloride (0.75-9.0 g/day), Group III: 30 with both CaCO(3) and sevelamer, Group IV: 15 with both CaCO(3) and cholestimide (1.5-6.0 g/day). These patients were prescribed several phosphate binders for at least 18 months. The serum levels of P, albumin-corrected Ca (Ca), Ca x P product and intact parathyroid hormone (iPTH) were serially determined. In Group I and IV, these four parameters showed no significant difference between at before administration and at after 1, 3, 6 and 12 months. In Group II, the values of iPTH, Ca and Ca x P product between at before sevelamer administration and at after 9 months were 199.43+/-94.40 vs 159.86+/-96.03 pg/mL (p<0.05), 9.48+/-1.12 vs 9.01+/-1.00 mg/dL (p<0.05) and 62.72+/-19.62 vs 50.44+/-25.97 mg(2)/dL(2) (p<0.05), respectively. In Group III, P showed significant decrease from 7.16+/ 1.33 to 6.72+/-1.69 mg/dL (p<0.05) between at the time of adding sevelamer to CaCO(3) and at after nine months. These results indicate that sevelamer plays an excellent role in the treatment of 2 degrees HPT mainly by controlling Ca level and the combination therapy with CaCO(3) is useful for improvement of P level in patients undergoing HD. PMID- 16272643 TI - [Effects of reduced dialysate calcium on mineral metabolism in hemodialysis patients]. AB - AIM: To study the effects of decreased dialysis calcium on mineral metabolism. METHODS: Dialysis calcium concentration was switched from 3.0 mEq/L to 2.5 mEq/L. Changes of serum Ca, P, and PTH were monitored for 6 months in 58 hemodialysis patients. RESULTS: Serum calcium decreased 2 weeks after the switch of dialysate, although it returned to the basal level after 6 months because of increased dosage of vitamin D. Phosphorus transiently increased after the switch. I-PTH increased in patients whose i-PTH before the switch was less than 100 pg/mL. PTH decreased in patients whose i-PTH exceeded 300 pg/mL. CONCLUSION: Decreased dialysis calcium produced lower serum calcium and better PTH control. PMID- 16272645 TI - [Control of calcium and phosphate in hemodialysis patients]. AB - In many hemodialysis patients at our hospital on May 2003, serum calcium levels were higher and intact PTH levels were lower than the target level of the K/DOQI guidelines. We could decrease doses of calcium-based phosphate binders by using sevelamer hydrochloride, and that could lower the level of calcium and raise the level of intact PTH. The number of patients who satisfy all the targets for calcium, phosphate, Ca x P and int-PTH, increased from 1.7% to 14.7% as a result of diet therapy and a change in their dose of medicine. It is natural that diet greatly influences on the control of calcium and phosphate, but if the period of hemodialysis is longer or there is swelling of the parathyroid, both diet therapy and medical therapy become more important for hemodialysis patients. PMID- 16272646 TI - [Significance of serum calcium and phosphorus control for the cardiac function in patients with long term hemodialysis]. AB - We evaluated how serum calcium and phosphorus will effect a cardiomegaly of hemodialysis patients. The cardiac valve of hemodialysis patients have a high incidence of calcification. Forty four percent of mitral valve and seventy five percent of aortic valve occurred the calcification, and the progress of calcification involve with increasing LVMI. The consequence of this results was that we consider prevent calcification is important to inhibiting factor of cardiomegaly, but serum calcium and phosphorus wasn't effect a cardiomegaly. On the other hand, we detected a significant negative correlation between serum calcium and LVMI (P = 0.0008) and a significant positive correlation between serum phosphorus and LVMI (P = 0.0105) . Consequently we thought that to control the serum phosphorus is important factor to inhibit the cardiomegaly. PMID- 16272647 TI - [Suitable examination intervals for hemodialysis patients with secondary hyperparathyroidism treated with maxacalcitol]. AB - Five patients with secondary hyperparathyroidism undergoing chronic hemodialysis, whose level of intact-PTH was less than 500 pg/mL, were treated with maxacalcitol for 16 weeks. The levels of serum intact-PTH, corrected calcium, phosphorous, ALP and BAP were measured every 2 weeks, and we investigated the suitable examination intervals to evaluate parathyroid function in these patients. Intact-PTH decreased significantly after 6 weeks, and thereafter corrected serum calcium level increased. From these results it is necessary to put emphasis to the time before 8 weeks in the measurement of intact-PTH, and on the other hand, after 8 weeks in the measurement of corrected serum calcium. ALP showed no significant change during the administration period. As a bone metabolic marker, BAP is more useful, and its measurement is enough for every 2 months. In conclusion, suitable examination intervals during the period of maxacalcitol treatment might not be adapted to the recommendation in the K/DOQI guideline 8B.2. PMID- 16272648 TI - [Acute response of serum PTH and bone markers after injection of 1alpha,25 (OH)2D3 and 22-oxacalcitrol in hemodialysis patient]. AB - Time-course changes of serum PTH and various bone markers were compared after injection of 1alpha,25 (OH)(2)D(3) with that of 22-oxacalcitriol (OCT) in hemodialysis patients. Five patients (M/F; 3/2, mean ages of 61.6 years) were enrolled into the present study. Oral administration of vitamin D(3) derivatives was stopped at least one week before initiation of vitamin injection. After 1 week of single intravenous injection of OCT (5 microg), 1alpha,25 (OH)(2)D(3) (0.5 microg) was followed. Serum levels of intact PTH, intact osteocalcin, bone alkaline phosphatase, cross-linked N-telopeptides of type I collagen, calcium, and phosphate were measured before, 24h, and 48 h after injections of vitamin D(3) derivatives. Significant difference did not exist in time-course changes of serum PTH, any of bone markers, calcium and phosphate between after OCT and 1alpha,25 (OH)(2)D(3) injection. In conclusion, the present study may not support the presence of significant direct effect of vitamin D(3) derivatives on bone metabolism in hemodialysis patients. PMID- 16272649 TI - [Effect of vitamin D on remaining kidney function after chronic kidney failure in preservation period patient]. AB - It is necessary to manage the secondary hyperparathyroidism at the early stage. We concern about vitamin D, one of the treatment for secondary hyperparathyroidism has some adverse effects on the kidney function. We administered oral vitamin D (alfa calcidol 0.25 microg/day) of 21 outpatients who have chronic renal failure of preservation period and observed clinical laboratory test result of serum creatinine, adjusted calcium, intact phosphate. Renal function is evaluated by the value of the decline of serum creatinine reciprocal. A small dosage of oral vitamin D may not effect on kidney function, but we should examine this though studying more cases. PMID- 16272650 TI - [Clinical significance of PTH (1-84) and PTH (7-84) in patients with predialysis chronic renal failure in relation to bone metabolism markers]. AB - Serum levels of parathyroid hormone (PTH) in predialysis patients with chronic renal failure (CRF) were measured using both the "intact PTH" and "bio PTH" assays, and serum levels of PTH (7-84) were assessed by subtracting bio PTH from intact PTH. The PTH values measured by the two assays were strongly correlated, and were also significantly positively correlated with both bone formation and resorption markers. PTH (7-84) was significantly positively correlated with both the intact PTH and bio PTH, and was also significantly positively correlated with the bone metabolism markers. There is no significant relationship between bio PTH/PTH (7-84) ratio and bone metabolism markers. In conclusion, bio PTH and intact PTH assays have similar clinical significance in predialysis CRF patients. The PTH (7-84) and even the ration of bio-PTH/PTH (7-84) have little specific clinical effect on bone metabolism. PMID- 16272651 TI - [Serum calcium levels at first renal replacement therapy and patients survival]. AB - In K/DOQI guideline, albumin adjusted serum calcium concentrations between 8.4 to 9.5 mg/dL and serum phosphorus concentrations between 3.5 to 5.5 mg/dL are recommended. But without clinical symptoms relating to hypocalcemia, medication for increasing serum calcium levels is not needed even less than 8.4 mg/dL. To clarify this guideline, we examined the data of 271 patients who started dialysis at Shinrakuen Hospital, their medication before renal replacement therapy and life prognosis. Medication made higher serum calcium concentrations (p< 0.005) at first renal replacement therapy. Concerning serum calcium concentrations, the lower group (<8.4 mg/dL) showed significantly better prognosis than the middle group (8.4 to 9.5 mg/dL) and the higher group (>9.5 mg/dL) (p< 0.05). When serum calcium levels were adjusted for the level of serum albumin, this tendency was stronger (p< 0.0001). We concluded that although serum calcium concentrations might not need control, nutritional states are far more important before starting dialysis. PMID- 16272652 TI - Benefits of maternal participation in newborn nurseries. AB - OBJECTIVE: To evaluate the outcome of active involvement of mothers/mother substitutes in day-to-day care of high risk neonates admitted in a level II newborn care unit. METHODS: An observational study was carried out over a period of eleven years incorporating active participation of mothers/substitute in the day to day care of their sick neonates. The outcome is assessed in terms of mortality due to the three major illnesses (asphyxia, sepsis and prematurity) during this phase. The data is compared with that of a similar level II care centre where conventional neonatal care is practised. RESULTS: There is a significant and sustainable reduction in neonatal mortality due to the three major illnesses when the mothers are also involved in the neonatal care, in spite of a considerable increase in the number of admissions during this period. CONCLUSION: The concept of active participation of mother/substitute in neonatal nursery ensures 1:1 care at all times. It is a cheap and effective alternative to inadequacy of bed:nurse ratio (BNR). PMID- 16272653 TI - Neonatal screening for glucose-6-phosphate dehydrogenase deficiency. AB - OBJECTIVE: This study was carried out to detect the incidence of erythrocytic Glucose-6 -Phosphate dehydrogenase (G-6-PD) deficiency, to compare the incidence of hyperbilirubinemia in G-6-PD deficient neonates as compared to G-6-PD normal neonates and to asses the usefulness of neonatal screening for G-6-PD deficiency. METHOD: In a retrospective hospital based study 2,479 male and female neonates consecutively born at Indraprastha Apollo hospital between July 1998 to June 2003 who were screened for G-6-PD levels were evaluated for the incidence of G-6-PD deficiency. RESULTS: Incidence of G-6-PD deficiency was found to be 2.0%. Incidence in males was 283% and female was 1.05%. The incidence of hyperbilirubinemia was found to be 32% in G-6-PD deficient neonates which was significantly higher than the incidence of hyperbilirubinemia in neonates with normal G-6-PD, which was 12.3% (P< 0.001). CONCLUSION: Our data suggests that neonatal screening for G-6-PD deficiency is a useful test for preventing and early treatment of complications associated with it. PMID- 16272654 TI - Infantile tremor syndrome in Iraqi Kurdistan. AB - OBJECTIVE: To describe a rare condition that occurred for the first time in Iraqi Kurdistan during the UN Sanctions on Iraq in the nireties. Literature review was made to fine possible causes. METHODS: Patients were collected from various sources including private clinics, hospital outpatient clinics and the Centre for Handicapped Children in the city of Arbil. Clinical features and proper history were the main diagnostic tools. Different treatment regimes were used on those patients. RESULTS: 24 patients were collected; the majority was between 6-18 months. Males were more affected than females. Half of the patients were exclusively breast-fed and three quarter have received blood for anemia. CONCLUSION: Infantile tremor syndrome is a rare condition which was described both in India and outside India as well. It was observed among children in Iraqi Kurdistan during the difficult economic situation under the UN Sanctions. The condition has the same features inside and outside India. The etiology is not very clear yet. PMID- 16272655 TI - Shunt revision in hydrocephalus. AB - OBJECTIVE: A retrospective analysis of 50 hydrocephalic children having a minimum follow-up of 6 months was carried out to see their etiology, clinical features, complications, incidence of shunt revisions, outcome and the variation from their Western counterparts. METHODS: Clinical features, image findings and treatment of all the cases were recorded from their discharge summaries. Record of shunt revision complications and outcome was maintained by the principal author. The data of all the cases were analyzed. RESULTS: The age of children varied from 1 month to 12 yr (mean 2.2 yr). The most common etiology of hydrocephalus was aqueductal stenosis in 18 (36%) children. Post infective hydrocephalus, either of post-tubercular meningitis (TBM) or following bacterial meningitis, remained the cause in 15 children (30%). Congenital TORCH infection was responsible for 3 cases of hydrocephalus making infective etiology as the cause in 18 (36%) cases. Intra 4th ventricular neurocysticercus cyst caused blockade of CSF pathway in 2 children. 15 out of 50 children required shunt revision, either due to infection (8,16%) or shunt obstruction (7, 14%). Multiple shunt revisions were required in 2 children only. These revisions were required due to infection, obstruction or malfunction of the shunt. CONCLUSIONS: Infective etiology is responsible for hydrocephalus in significant number of children (36%). The possibility of TORCH infection, as a cause of hydrocephalus should be considered even amongst the children of screened mothers during antenatal check-up. Pure intra 4th ventricular neurocysticercus cysts (without intraparenchymal cyst), though rare, can manifest with outlet obstruction. Incidence of shunt revision using Chhabra's medium pressure shunt is very high in children at an average follow up of 1.6 yr. Post infective hydrocephalus is a major cause of delayed milestones, contributing to mental retardation. PMID- 16272656 TI - Therapeutic potentials of bovine colostrums. AB - Colostrum is the first milk produced by mammals for their young ones. This transfers the passive immunity gained by the mother to the baby. The bovine colostrum (BC) can be obtained in large quantity and has properties similar to human colostrum. It has been used for various disorders of the body. It has properties to stimulate immune system, contains growth factors and many bioactive substances needed for the body to combat with wear and tear. The BC has been used for various gastrointestinal disorders, respiratory tract infection, rheumatoid arthritis, healing injured tissues of body etc. There are not much double blind placebo-controlled trials to prove its efficacy, though a lot of experience about its good effects in various disorders is available in the literature. The dosage and duration of therapy need to be worked up. The BC has potential to treat as well to prevent certain diseases in the body. In future this will prove to be a very useful product to treat and control diseases in a natural way. PMID- 16272657 TI - Developmental and behavioral pediatrics--the present and the future. PMID- 16272658 TI - Neuroplasticity in children. AB - Research in the field of neurosciences and genetics has given us great insight into the understanding of learning and behavior and changes in the brain in response to experience. It is seen that brain is dynamically changing throughout life and is capable of learning at any time. Critical periods of neuroplasticity for various streams of development are also better understood. Technological advances in non invasive imaging techniques and advances in molecular genetics have helped us understand the basis of many developmental disorders which may help in planning effective intervention strategies. PMID- 16272659 TI - Syndromes and disorders associated with mental retardation. AB - This article reviews selective comments on the concept of Mental Retardation (MR) in adolescents. Issues covered include the definition, prevalence, and differential diagnosis of MR. Some of the syndromes and disorders associated with MR in the adolescents are also considered with emphasis on the behavioral concerns that may be present in this age group. Finally, concepts of management by the clinician are reviewed. It is recommended that health care professionals caring for adolescents with MR should help these youths maximize their potential as human beings, helping them achieve meaningful functioning in adulthood. PMID- 16272660 TI - Cerebral palsy-definition, classification, etiology and early diagnosis. AB - Cerebral palsy is a common neurodevelopmental condition encountered by pediatricians. The condition may present itself in many different clinical spectra. The etiological and risk factors are many and an awareness of the interplay of multiple factors in the causation of CP is crucial. In many cases, the cause of Cerebral palsy may not be apparent. Cerebral palsy is invariably associated with many deficits such as mental retardation, speech and language and oromotor problems. A thorough neurodevelopmental assessment of the child with Cerebral Palsy should include evaluation of associated deficits so that a comprehensive early intervention program an be planned and executed. PMID- 16272661 TI - Pharmacologic interventions for reducing spasticity in cerebral palsy. AB - Motor function abnormalities are a key feature of cerebral palsy. Spasticity is one of the main motor abnormalities seen in children with cerebral palsy. Spasticity is a velocity dependent increased resistance to movement. While in some children, spasticity may adversely impact the motor abilities, in others, it may help maintain posture and ability to ambulate. Thus, treatment to reduce spasticity requires careful consideration of various factors. Non-pharmacologic interventions used to reduce spasticity include physiotherapy, occupational therapy, use of adaptive equipment, various orthopedic surgical procedures and neurosurgical procedures. Pharmacologic interventions used for reducing spasticity in children with cerebral palsy reviewed in this article include oral administration of baclofen, diazepam, dantrolene and tizanidine, intrathecal baclofen, and local injections of botulinum toxin, phenol, and alcohol. PMID- 16272662 TI - Kawasaki disease: are we missing the diagnosis? AB - We report 6 cases of Kawasaki disease (KD) diagnosed over a period of one year and review of all the cases reported from India. The diagnosis of KD was based on clinical criteria The mean age of patients was 6.83 years and mean duration of symptoms before diagnosis was 7.5 days. Apart from classical clinical features, elevated transaminases and blood urea along with free fluid in abdomen was present in one case each. Two patients had dilated coronaries that returned to normal on follow up. One patient developed headache and neck stiffness following treatment with intravenous gamma globulins. The outcome was excellent in all the cases. PMID- 16272664 TI - Factors affecting antibiotic prescribing pattern in pediatric practice. AB - To determine the proportion of children receiving antibiotics for common illnesses and to understand the antibiotic prescription ptern and factors influencing it, a cross sectional study was done among the private practitioners in Chennai, India 403 prescriptions by 40 physicians from selected health facilities were analyzed 79.9% of children with ARI (Acute respiratory infection) and ADD (Acute watery diarrhea) were prescribed antibiotics. Penicillins (43.9%) were the commonest antibiotic prescribed. Factors like postgraduate qualification, experience of physician, source and method of updating knowledge, inpatient practice setting and presence of fever influenced the antibiotic prescription. PMID- 16272665 TI - Keratomalacia in a neonate secondary to maternal vitamin A deficiency. AB - A 40-day-old male child was admitted with complaints of not opening eyes from 2-3 days after birth. A diagnosis of X3-B keratomalacia was made. The treatment was done with vitamin-A to which the patient had responded. The mother of the baby had a history of night blindness throughout the pregnancy for which she was also treated. Keratomalacia secondary to vitamin-A deficiency is rare in neonates, although in children it is reported form developing countries. PMID- 16272666 TI - Scurvy in infantile tremor syndrome. AB - Infantile Tremor Syndrome is a distinct clinical entity most commonly seen in Indian Subcontinent. Syndrome consists of tremors, mental and developmental retardation, abnormal skin pigmentation and anemia in children between 6 months to 2 years. The etiology is still elusive. Amongst various theories, nutritional theory is the most accepted. So far there are no cases reported of vitamin C deficiency in ITS. In this article, three cases of ITS associated with vitamin C deficiency are reported. PMID- 16272667 TI - Congenital chylothorax treated with octreotide. AB - Congenital chylothorax is the accumulation of lymphatic fluid within the pleural space. Cases unresponsive to conservative management usually require surgery. Octreotide has been used successfully to treat post-traumatic chylothoraces in the pediatric and adult population. Its exact mode of action is uncertain but it is believed to reduce lymphatic drainage by a direct action on splanchnic lymph flow. We report a case of congenital chylothorax where surgery was avoided with the compassionate trial of the somatostatin analogue, octreotide. A 33 week gestation female infant, born with the presence of large bilateral pleural effusion, was unresponsive to conservative management. Octreotide was commenced on day 15, with 10 days of an octreotide infusion, initially 0.5 microg/kg per hour and increased daily by 1 microg/kg per hour to 10 microg/kg per hour. Treatment was associated with prompt respiratory improvement prior to cessation of pleural drainage over the 10 day. She remains well at 6 months of age. Further studies are required to ascertain its true value in congenital chylothorax. PMID- 16272668 TI - Tumoral calcinosis with hyperphosphatemia. AB - Tumoral calcinosis is a rare disorder of mineral metabolism among adolescents and young adults characterized by deposition of calcific masses around large joints. It is less commonly reported in pediatric population and commonly mistaken for bone tumors. Typical lab parameters include hyperphosphatemia with normal levels of serum calcium, parathyroid hormone (PTH) and alkaline phosphatase. A ten-year old boy with typical features of tumoral calcinosis is presented. PMID- 16272670 TI - Measles outbreak in a migrant population. PMID- 16272669 TI - Menkes' kinky hair syndrome. PMID- 16272671 TI - Maternal acute renal failure and non-immune hydrops. PMID- 16272673 TI - BCG under universal immunization programme. PMID- 16272675 TI - Superspecialty selection in postgraduate education. PMID- 16272676 TI - Biochemical measures in the diagnosis of alcohol dependence using discriminant analysis. AB - BACKGROUND: Alcohol dependence often cannot be diagnosed based on self-report alone. Various biochemical and haematological parameters have been used to screen alcohol use disorders. AIM: To develop discriminant equations based on lipid and liver measures independently for identifying alcohol dependent and non-dependent subjects. SETTINGS AND DESIGN: Case control study in a tertiary care hospital. METHODS AND MATERIALS: One hundred subjects fulfilling the criteria of alcohol dependence and seventy healthy controls were included. The socio-demographic details, caloric intake, height, weight and blood pressure were recorded. Samples were analysed for various lipid measures as well as liver function. STATISTICAL ANALYSIS USED: Diagnostic values such as sensitivity, specificity, positive predictive value (PV+), negative predictive value (PV-) and discriminant analysis. RESULTS: Using discriminant analysis, two equations were constructed based on liver and lipid measures independently. 84.7% of the subjects on the basis of total cholesterol (TC), apolipoprotein B (ApoB) and low density lipoprotein/high density lipoprotein-cholesterol (LDL/HDL-c and 89.1% on the basis of aspartate amino transferase (AST) and gamma glutamyl transferase (GGT) were correctly classified into their respective groups. CONCLUSIONS: This study demonstrates the ability of TC, ApoB and LDL/HDL-c (among lipid measures) and AST and GGT (among liver measures) in discriminating alcohol dependents from non dependent subjects. PMID- 16272678 TI - Interlocking nailing of humeral shaft fractures. A retrospective study of 114 patients. AB - BACKGROUND: Fractures of the humeral shaft are relatively common injuries. Literature suggests that humeral shaft fractures represent approximately 3 % of all fractures. There are several modalities for the management of diaphyseal humeral fractures. The latest investigations emphasize the concept of minimal exposure and rigid fixation. AIM: The aim of the study is to evaluate the results of antegrade intramedullary nailing in humeral shaft fractures. DESIGN: A retrospective review. SETTINGS: Patients were treated in private hospital settings by 3 orthopaedics surgeon. MATERIAL AND METHODS: Between 1995 and 2003, the technique of antegrade locked intramedullary nailing with UHN in humeral shaft fractures was performed on 114 patients. Forty-two (36%) patients sustained multiple traumas, and 22 (19%) fractures were open. The outcomes were evaluated with a mean follow-up of 41 months. STATISTICAL ANALYSIS USED: Ranges of results given. RESULTS: In 109 fractures primary union observed. In the other five patients union achieved after removal of the nail and fixation with DCP and bone grafting. The average time for union was 13 weeks (range, 10-36 weeks). One hundred-five patients had excellent or satisfactory recovery of shoulder and elbow function. Complications included impingement due to proximal locking screws in two patients and prominent nail in three patients, transient postoperative radial nerve palsy in four patients. CONCLUSIONS: This study shows that antegrade locked nailing in humeral shaft fractures are reliable and also effective in multiply injured patients. PMID- 16272677 TI - Angiotensin converting enzyme gene polymorphism in Indian children with steroid sensitive nephrotic syndrome. AB - CONTEXT: Nephrotic syndrome is one of the commonest renal problem encountered in children. It is difficult to predict at onset, the clinical course in terms of steroid responsiveness or resistance. Angiotensin converting enzyme (ACE) gene insertion/deletion (I/D) polymorphism has been studied as a predictor of clinical course in common multi-factorial diseases including focal segmental glomerulosclerosis. There is no study available from our country till date to find out any correlation of the steroid response in idiopathic nephrotic syndrome and ACE gene polymorphism. AIM: To study distribution and correlation of ACE gene I/D polymorphism in idiopathic steroid sensitive nephrotic syndrome. SETTINGS & DESIGN: Case-control retrospective study. SUBJECTS & METHODS: We studied ACE gene polymorphism in 90 consecutive patients (82% males, 18% females) with steroid sensitive idiopathic nephrotic syndrome and 300 normal controls (NC). STATISTICAL ANALYSIS: Chi-square test and Fisher's exact test (for cases with insufficient expected cell frequencies). RESULTS: The mean age of onset was 5.3 +/- 4 years. Steroid sensitive (SS) patients showed II (SS-48%, NC-26%) genotype was more frequent than normal controls (p=0.002). There was no significant difference in genotype frequencies among steroid SS subgroups. CONCLUSIONS: In our study II genotype was more frequent in steroid sensitive nephrotic syndrome children in comparison to normal controls. Further functional studies with large number of children are required to investigate the role of II genotype in steroid sensitive nephrotic syndrome. Comparison of the genotypic frequency with steroid resistant patients may provide information, which might be useful in clinical practice. PMID- 16272680 TI - Acquired angioedema secondary to hormone replacement therapy. AB - Angioedema is a potentially life threatening condition and may be either inherited or acquired. The latter is rare with only a handful of cases reported in the world literature. Presenting complaints are often vague. Those most commonly described include swelling in the subcutaneous and submucosal tissues. Patients presenting with laryngeal edema have high mortality, and high clinical suspicion is necessary to avoid instrumentation, which can precipitate laryngeal spasm. We present a review of reported cases of hormonally induced hereditary angioedema, along with a report of a patient with acquired angioedema secondary to hormone replacement therapy. To the best of our knowledge, this case probably represents the first reported case of acquired angioedema secondary to hormone replacement therapy. PMID- 16272679 TI - HIV-1 western blot assay: What determines an indeterminate status? AB - BACKGROUND: The Western blot assay is the gold standard for the detection of antibodies to human immunodeficiency virus type 1 (HIV-1). However, indeterminate Western blot reactivity to HIV-1 proteins may occur in individuals, who may not be infected with HIV. AIM: This retrospective study was aimed to determine the diagnostic value of the interpretation criteria in relation to commercial kits for HIV-1 diagnosis. METHODS AND MATERIALS: A total of 556 serum/plasma specimens collected from high-risk population attending our HIV clinic from 2000-2004 were tested by three different western blot kits: NEW LAV BLOT I (n=244), HIV BLOT 2.2; (n=112), Genetic Systems HIV-1 (n=237). And the results of western blot strips were analyzed using the various interpretation criteria: WHO/NACO, CDC/ ASTPHLD, ARC, FDA, CRSS and JHU. Some specimens were run on more than one kit. RT PCR assay was performed on 5 specimens, which were indeterminate with LAV BLOT I. RESULTS: The discrepancy in LAV BLOT I positive results were between 157(64) 176(72), and indeterminate results were between 44(18) to 63(25). No such variations were observed in genetic systems. There are some HIV negative (by PCR) specimens were indeterminate in LAV BLOT I revealing the kit more sensitive and less effective for diagnostic purpose. CONCLUSION: The genetic systems kit is superior to other kits we analyzed and its results are concordant with HIV-1 PCR results. To report, the choice of western blot commercial kit is paramount important than the use of particular interpretation criteria for the diagnosis of HIV-1. PMID- 16272682 TI - The cocaine 'body-packer' syndrome: diagnosis and treatment. PMID- 16272681 TI - Parasites (filaria & strongyloides) in malignant pleural effusion. PMID- 16272683 TI - An elderly man with dysphagia and dysarthria. PMID- 16272684 TI - In vivo multitracer analysis technique: screening of radioactive probes for noninvasive measurement of physiological functions in experimental animals. AB - A novel screening experiment, to find radioactive probes for non-invasive measurements of physiological functions in experimental animals, was tested using the in vivo multitracer analysis technique. The details of the efficiency of the detector settings used in the in vivo multitracer analysis technique were examined by both computer simulations and practical measurements. Multiple radioactive isotopes, i.e. multitracer, were prepared by irradiating a silver foil target with a heavy ion beam at the RIKEN ring cyclotron. After chemical separation of the silver target, the multitracer was finally dissolved in isotonic citrate buffer. The multitracer solution was intravenously injected into rats. Using a gamma-ray detector equipped with a well-defined slit, the collimated gamma-rays from the upper abdomen of living rats were measured. After correction of detection efficiencies, it was possible to compare the distribution of radioactive elements between two groups of rats different in body weight. The in vivo measurement showed that the tissue substantial volume of the selenium deficient (SeD) rat liver increased compared to normal rats. The possibility of a functional estimation of tissue/blood volume for living rats was proposed based on the characteristic in vivo distribution of 74As, 83Rb and 103Ru. PMID- 16272685 TI - Two proteins, Mn2+, and low molecular cofactor are required for C-glucosyl cleavage of mangiferin. AB - C-Glucosides, in which sugars are attached to the aglycone by carbon-carbon bonds, are generally resistant to acid and enzyme hydrolysis. The C-glucosyl bond of mangiferin, a xanthone C-glucoside, was cleaved by anaerobic incubation with a human intestinal bacterium, Bacteroides sp. MANG, to give norathyriol. A cell free extract obtained by sonication of B. sp. MANG demonstrated cleaving activity for mangiferin to norathyriol by adding NADH, diaphorase, and dithiothreitol. Both high molecular weight (>10 k) and low molecular weight (<10 k) fractions obtained from the cell-free extract were required for the activity. MnCl2 was necessary for the activity, but other metal ions were not. By purification of the high molecular weight fraction using DEAE-cellulose and Phenyl Sepharose column chromatography, two fractions, designated as proteins A and B, were separated and required for the activity. Neither protein A nor protein B alone showed any activity. This is the first report describing a C-glucosyl-cleaving enzyme from human intestinal bacterium that seems to involve a novel enzyme mechanism. PMID- 16272686 TI - Insulin signaling in adipocytes differentiated from mouse stromal MC3T3-G2/PA6 cells. AB - The stromal MC3T3-G2/PA6 (PA6) cells from mouse clavaria did not require insulin for differentiation into mature adipose cells, although insulin is well known to play a key role in adipocyte differentiation. Large lipid droplets were observed in the cytoplasm of PA6 cells, and mRNA expression of the adipose specific proteins (aP2, PPARgamma, C/EBPalpha, FAS, GLUT4, leptin, and adiponectin) as differentiation markers appeared or increased clearly in the cells at 8 d after stimulation without insulin. In addition, the glycerol released from the cells (lipolysis) was increased in a concentration-dependent manner by isoproterenol. However, the isoproterenol-induced lipolysis in the cells was not influenced by treatment with insulin, although that was observed in extramedullary adipocytes, 3T3-L1 cells. On the other hand, the 2-deoxy-D-[1-3H]glucose uptake in differentiated PA6 cells also increased by insulin, as shown in other adipose cells. In the cells, insulin induced the phosphorylation of extracellular signal regulated kinases (Erks), Akt at Ser 473 and ribosomal p70 S6 protein kinase (p70 S6K) at Thr 389, and the insulin-induced 2-deoxy-D-[1-3H]glucose uptake was inhibited by pre-treatment with wortmannin, an inhibitor of phosphatidylinositol 3-kinase (PI3K), or ML-9, an Akt inhibitor. These results suggest that the insulin signal for adipogenesis (lipogenesis) and lipolysis in bone marrow stroma PA6 cells differs from extramedullary adipocytes, such as 3T3-L1 cells. PMID- 16272688 TI - Tissue-specific and de novo promoter methylation of the mouse glucose transporter 2. AB - Glucose transporter 2 (GLUT2) is tissue-specifically expressed in liver and kidney, and reduced in neoplastic hepatic lesions and in most hepatoma cell lines. Here we examined the involvement of epigenetic modifications in the regulation of GLUT2. Four CpGs in the GLUT2 promoter were undermethylated in GLUT2-expressing tissues. In isolated hepatocytes, GLUT2 expression declined and the promoter was methylated de novo. This de novo methylation occurred with a similar time-course in hepatocytes cultured in a high-glucose medium that induced GLUT2 expression, suggesting that de novo methylation can be induced independently of GLUT2 expression. GLUT2 was reactivated in hepatocytes following exposure to the methylation inhibitor 5-aza-2'-deoxycytidine (AzaC) but only after the methylation had occurred. In p53-deficient mouse liver, the CpGs were methylated de novo; the GLUT2 expression declined. The GLUT2 promoter was hypermethylated in Hepa1c1c7 cells, but expression could be rescued by AzaC. Thus, it is proposed that DNA methylation has an important role in the regulation of GLUT2 in mouse tissues and liver-derived cells. PMID- 16272687 TI - Intestinal gene expression in TNBS treated mice using genechip and subtractive cDNA analysis: implications for Crohn's disease. AB - So far it has proven difficult to identify a causative gene(s) or gene product initiating the events that lead to inflammation of the intestinal mucosa and, ultimately, progression to Crohn's disease (CD), an inflammatory bowel disease. However, gene transcripts identified in the intestine of trinitrobenzene sulfonic acid (TNBS)-treated mice might suggest a clue, and even represent candidate genes leading to inflammation and mucosal damage, and to subsequent fibrosis. In the present study, DNA microarray (13000 transcripts) methodology was applied to mucosal RNA extracted from TNBS-treated mice, some transcripts of which were validated via cDNA subtraction and RT-PCR analyses. Intestinal biopsy samples from CD patients were then analyzed using cDNA mini-array (1300 cDNAs), focusing on gene transcripts associated with cancer and immunity. Mini-array results revealed transcript changes similar and also dissimilar to those found from the DNA microarray analysis. These changes, previously known or newly identified, possibly occurring during the initial and progressive stages of inflammatory conditions may provide a clue to identify marker transcripts and/or targets for the development of future gene therapy. PMID- 16272689 TI - Expression of constitutive androstane receptor splice variants in rat liver and lung and their functional properties. AB - The mammalian constitutive androstane receptor (CAR) is a transcription factor that participates in controlling the expression of xenobiotic metabolizing and transporting genes in response to xenobiotics in an organ-specific manner. In addition to the wild-type CAR (CAR WT) mRNA, mRNAs for five splice variants (SVs) could be detected in the liver of 7-week-old male Wistar rats by RT-PCR using primer pairs covering a full-length mRNA derived from 9 exons; insertion of 18 bp at the 5'-end of intron 8 with or without deletion of 3 bp from the 5'-end of exon 7 (CAR SV1 or SV2), deletion of 4 bp from the 5'-end of exon 8 (CAR SV3), insertion of 195 bp intron 7 (CAR SV4), and insertion of 91 bp intron 6 (CAR SV5). In contrast, only CAR SV5 was detected in lung. Due to the introduction of novel stop codons, all the SVs were considered to code for premature proteins. The liver homogenate gave two protein bands in the vicinity of 37 kDa on Western blotting. They were attributable to CAR WT and SV-complex, respectively, based on their putative molecular weights in descending order. Upon cotransfection with the reporter plasmid, only the cells transfected with the CAR SV4-expression plasmid showed enhanced luciferase activity similar to the WT-transfected cells, for which the further splicing of the remaining intron 7 seemed to be responsible. The transactivation-defective SVs downregulated CAR WT-induced luciferase activity to some extent in the cotransfection experiments. PMID- 16272690 TI - Inhibition of benzo(a)pyrene induced lung adenoma by panax ginseng extract, EFLA400, in Swiss albino mice. AB - In the present investigation the chemopreventive action of Panax ginseng extract, EFLA400, in Swiss albino mice has been evaluated. We used a 9-week medium term anticarcinogenicity test model of lung adenomas [Yun et al.1)]. Lung adenomas were induced by single subcutaneous injection in the subscapular region with 0.02 ml of benzo(a)pyrene (BP) (0.5 mg suspension in 1% aqueous gelatin) in newborn mice (less than 24 h old). Also chromosomal aberrations and micronuclei induction were evaluated in bone marrow cells. These genotoxicity end-points were compared with adenoma incidence at the same dose levels of BP and EFLA400. The oral administration of EFLA400 (10 mg/kg body weight) showed significant reduction in number of adenomas and weight of the lungs induced by BP. A significant reduction (p<0.001) in lung adenoma incidence in EFLA400-treated mice was observed as compared to the 68.3+/-2.96% lung adenoma incidence in BP-alone group. The inhibition rate was 72.05+/-1.36% in EFLA400-treated group with respect to the reference group (BP-alone group). However, tumor multiplicity was observed as 0.91+/-0.08 and 0.25+/-0.01 in BP alone and BP+EFLA400-treated groups respectively. In EFLA400-treated group significantly reduced frequencies of chromosomal aberrations and micronuclei induced by BP were observed. The results of the present investigation suggest the chemopreventive action and antimutagenic effect of EFLA400 in Swiss albino mice induced by BP in newborn mice. PMID- 16272691 TI - Bcl-2 up-regulation and P-p53 down-regulation account for the low sensitivity of murine L929 fibrosarcoma cells to oridonin-induced apoptosis. AB - Drug resistance has been a major limitation to chemotherapy. There are many mechanisms that contribute to such resistance. In our study, we subcloned oridonin-sensitive and low sensitive L929 cells and both types of cells grew at almost the same growth rate. The acquired low sensitivity to oridonin-induced apoptosis was associated with Bcl-2 up-regulation and down-regulation of p53 phosphorylation. The p38 inhibitor SB203580 decreased Bcl-2 expression in the low sensitive L929 cells and made the cells more sensitive to oridonin. Moreover, a higher dose of oridonin promoted p53 phosphorylation, increased Bax expression and subsequently induced death of low sensitive L929 cells, however, it had no effect on Bcl-2 expression. The increased Bcl-2/Bax ratio in oridonin low sensitive L929 cells did not inhibit caspase-9 or -3 activation, but suppressed the cleavage of poly (ADP-ribose) polymerase (PARP), indicating the existence of caspase-9 or -3 independent PARP activation. These results indicated that in L929 cells, there was a relationship among the low sensitivity to oridonin, down regulation of p53 phosphorylation and Bcl-2 up-regulation. PMID- 16272692 TI - Enzymologic and pharmacologic profile of loxoprofen sodium and its metabolites. AB - We investigated the mechanism of inhibition of loxoprofen sodium, a non-steroidal anti-inflammatory drug (NSAID), and its active metabolite (loxoprofen-SRS) on cyclooxygenase (COX). In in vitro assays, loxoprofen sodium appeared inactive against recombinant human COX-1 and COX-2, whereas loxoprofen-SRS inhibited both. In the investigation of kinetic behavior, loxoprofen-SRS showed time-dependent inhibition for both isozymes. Human whole blood assay also showed that loxoprofen SRS possesses the profile of a non-selective inhibitor for COX. In a rat air pouch model, oral administration of loxoprofen sodium lowered prostaglandin (PG) E2 in both fluid exudates of the inflammatory pouch and stomach tissue with ED50 values of 2.0 and 2.1 mg/kg, respectively. Additionally, platelet thromboxane B2 production was also inhibited by loxoprofen sodium (ED50 of 0.34 mg/kg). In a rat carrageenan-induced paw edema model, loxoprofen sodium dose-dependently reduced the paw edema, accompanied by a decrease in PGE2 content in inflamed paw exudates. These findings suggest that the COX inhibitory activity of loxoprofen sodium is attributable to its active metabolite, loxoprofen-SRS, and that loxoprofen-SRS shows non-selective inhibition for COX. PMID- 16272693 TI - Effect of combined vitamin E and insulin administration on renal damage in diabetic rats fed a high cholesterol diet. AB - In the present study, we investigated the effects of a long-term treatment with vitamin E, an antioxidant vitamin, insulin, or their combination on renal damage in streptozotocin (STZ)-induced diabetic rats fed a high cholesterol diet. Increases in urinary albumin and lipid peroxide (LPO) excretions were observed in these diabetic rats, when both urinary parameters were measured at 8 and 15 weeks after STZ administration. Daily treatment with vitamin E, insulin, or their combination markedly suppressed the increase in the 24 h urinary albumin and lipid peroxide excretions. Furthermore, glycogen degeneration of distal tubules, fatty degeneration of glomerular endothelium and hypertrophy of glomeruli and mesangium were observed in the kidneys of the diabetic animals when histopathological evaluation was performed at 4, 8, and 15 weeks (glomerular and mesangial hypertrophy were observed only at 15 weeks). Combined vitamin E and insulin treatment was the most effective at suppressing these renal histopathological changes. These results indicate that combined vitamin E and insulin treatment additively prevents the development and progression of renal damage in diabetic rats. Possible mechanisms for the preventive effect of this combined treatment are discussed. PMID- 16272694 TI - Protective effect of serum thymic factor, FTS, on cephaloridine-induced nephrotoxicity in rats. AB - Serum thymic factor (FTS), a thymic peptide hormone, has been reported to increase superoxide disumutase (SOD) levels in senescence-accelerated mice. In the present study, we examined the effect of FTS on cephaloridine (CER)-induced nephrotoxicity in vivo and in vitro. We previously reported that CER led to extracellular signal-regulated protein kinase (ERK) activation in the rat kidney. So, we also investigated whether FTS has an effect on ERK activation induced by CER. Treatment of male Sprague-Dawley rats with intravenous CER (1.2 g/kg) for 24 h markedly increased BUN and plasma creatinine levels and urinary excretion of glucose and protein, decreased creatinine clearance and also led to marked pathological changes in the proximal tubules, as revealed by electron micrographs. An increase in phosphorylated ERK (pERK) was detected in the nuclear fraction prepared from the rat kidney cortex 24 h after CER injection. Pretreatment of rats with FTS (50 microg/kg, i.v.) attenuated the CER-induced renal dysfunction and pathological damage. FTS also suppressed CER-induced ERK activation in the kidney. In vitro treatment of the established cell line, LLC PK1 cells, with FTS significantly ameliorated CER-induced cell injury, as measured by lactate dehydrogenase (LDH) leakage. Our results, taken together with our previous report that MEK inhibitors ameliorated CER-induced renal cell injury and ERK activation induced by CER, suggest that FTS participates in protection from CER-induced nephrotoxicity by suppressing ERK activation induced by CER. PMID- 16272695 TI - HMG-CoA reductase inhibitors do not improve glucose intolerance in spontaneously diabetic Goto-Kakizaki rats. AB - We examined whether 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) improve glucose intolerance in spontaneously diabetic Goto Kakizaki (GK) rats or not. The fasting blood glucose, plasma insulin, and serum cholesterol levels were significantly higher in GK rats than those in age-matched Wistar rats. All rats were given orally once a day 0.5% carboxymethylcellulose, pravastatin 8 mg/kg, simvastatin 8 mg/kg, or atorvastatin 8 mg/kg. An oral glucose tolerance test (OGTT) was performed before and 3, 6 and 12 weeks after statin treatments. The hyperglycemic response to OGTT in GK rats significantly exceeded that in Wistar rats. The plasma insulin level in GK rats increased with age until 14-week-old (treated for 6 weeks), and then decreased. Glucose intake significantly increased the plasma insulin in almost all rats. The increment of plasma insulin due to OGTT in GK rats appeared to be less than that in Wistar rats, because the basal level was already high in GK rats. Pravastatin, simvastatin, and atorvastatin did not modify changes in blood glucose and plasma insulin induced by glucose intake. In conclusion, long-term treatments of GK rats with statins did not improve glucose intolerance observed during OGTT. PMID- 16272696 TI - YM-231146, a novel orally bioavailable inhibitor of vascular endothelial growth factor receptor-2, is effective against paclitaxel resistant tumors. AB - Chemotherapy using anticancer drugs induces serious the problem of multidrug resistance (MDR) in the cancer cells. In contrast, endothelial cells so rarely acquire MDR that antiangiogenesis therapy has recently been considered as an effective means for cancer chemotherapy. We screened compounds in the chemical library to find a novel and orally active antitumor agent with vascular endothelial growth factor receptor-2 tyrosine kinase (VEGF-R2 TK) inhibition. The result was YM-231146 (IC50=0.080 microM). YM-231146 inhibited VEGF-stimulated proliferation, VEGF-R2 autophosphorylation, and vessel sprout formation of human vascular endothelial cells at concentrations between 0.15-0.30 microM. However, YM-231146 did not inhibit cancer cell proliferation at these concentrations (IC50>5 microM). In the in vivo studies, once-daily oral dosing of YM-231146 to human cancer xenografts elicited antitumor activity at doses of 3-100 mg/kg. Moreover, YM-231146 completely inhibited tumor growth of paclitaxel-resistant cancer cells without decreasing body weight at a dose of 100 mg/kg. These results suggest that YM-231146 is a novel orally bioavailable inhibitor of VEGF-R2 that would be useful for the treatment of multidrug resistant tumors. PMID- 16272697 TI - Transformation of ginsenosides Rb2 and Rc from Panax ginseng by food microorganisms. AB - Protopanaxadiol ginsenosides Rb2 and Rc were transformed using cell-free extracts from various edible food microorganisms and then analyzed by TLC and HPLC. Rb2 and Rc were transformed into compound K via Rd and F2 by Bifidobacterium sp. Int57 and Bifidobacterium sp. SJ32. Lactobacillus delbrueckii transformed Rb2 and Rc into ginsenoside Rh2. Bifidobacterium sp. SH5 transformed Rb2 and Rc into F2. Aspergillus niger transformed Rb2 into compound K via compound O and compound Y, whereas it transformed Rc into compound K via Mc. Taken together, these processes would allow a specific bioconversion process to obtain specific ginsenosides using an appropriate combination of ginsenoside substrates and specific microbial enzymes. PMID- 16272698 TI - Antihyperlipidemic effect of crocin isolated from the fructus of Gardenia jasminoides and its metabolite Crocetin. AB - The pancreatic lipase inhibitors were isolated from the fructus of Gardenia jasminoides ELLIS, and their antihyperlipidemic activities were measured. Gardeniae fructus (GF) water extract inhibited pancreatic lipase activity. Crocetin and crocin were isolated from GF water extract as inhibitors of pancreatic lipase with an IC50 value of 2.1 and 2.6 mg/ml (triolein as a substrate). Crocin and crocetin significantly inhibited the increase of serum TG level in corn oil feeding-induced triglyceridemic mice, as well as that of serum triglyceride and total and LDL cholesterol levels in Triton WR-1339-induced hyperlipidemic mice. These compounds also showed hypolipidemic activity in hyperlipidemic mice induced by high cholesterol, high fat or high carbohydrate diets for 5 weeks. The results suggest that the hypolipidemic activity of GF and its component crocin may be due to the inhibition of pancreatic lipase and crocin, and its metabolite, crocetin, can improve hyperlipidemia. PMID- 16272699 TI - Anti-angiogenic and immunomodulatory effect of the herbal medicine "Juzen-taiho to" on malignant glioma. AB - Juzen-taiho-to (JTT) is known as a Japanese herbal medicine that increases the immune function via the enhancement of phagocytosis, cytokine induction, and antibody production. Anti-neoplastic effects on malignant gliomas have been reported by means of the enhancement of the immune function in both animals and humans. We evaluated whether JTT has anti-angiogenic effects on malignant glioma growth in vitro and in vivo. In vitro, the anti-proliferative effect of JTT on malignant glioma cells and endothelial cells was assessed by cell proliferation assay. In vivo, a subcutaneous model of malignant glioma with different-aged mice (old, 43 weeks; young, 8 weeks) was used. After oral administration of JTT to mice, their immunological function and angiogenic status of tumor tissues were assessed by flow cytometry and immunohistochemistry, respectively. JTT inhibited human endothelial cell, but not glioma cell, proliferation in vitro. In vivo, the NK (natural killer) cell ratio within PBMC (peripheral blood mononuclear cells) and NK activity of fresh splenocytes obtained from JTT-treated old mice were significantly increased compared to the ratio and activity in control mice. In old mice, the vessel area of tumor tissues in JTT treatment groups was significantly decreased. These enhancements of immunological function and the inhibition of angiogenic activity were not observed in young mice. JTT not only increased host immunological function but also exerted anti-angiogenic effects on malignant glioma growth. JTT would be useful as an adjuvant medicine for malignant gliomas through its enhancement of systemic immunological function and its anti-angiogenic action. PMID- 16272700 TI - Thiopurine methyltransferase genotype and phenotype status in Japanese patients with systemic lupus erythematosus. AB - We investigated the genotypic status of thiopurine methyltransferase (TPMT) polymorphism to evaluate the possible risk of the toxicity of azathioprine (AZA) in 68 patients with systemic lupus erythematosus (SLE). The allele frequency of TPMT mutation in the SLE group (2.9%) was higher than that in 174 Japanese healthy volunteers (1.1%), although it did not reach statistically significant difference (p=0.23). The mean value of TPMT activities in 51 subjects with TPMT*1/*1 was 40% higher than that of 4 subjects with TPMT*1/*3C in SLE group (18.1+/-6.1 nmol/h/ml packed red blood cells (pRBC) versus 13.2+/-3.2 nmol/h/ml pRBC; p=0.11). Two out of 4 SLE patients with TPMT*1/*3C had been treated with AZA, and one patient showed a leucopenia. The TPMT genotyping before AZA treatment is recommended for Japanese SLE patient group to avoid the AZA-induced adverse events, although detection of the patient with low TPMT activity by genotyping is still imperfect. PMID- 16272701 TI - Effect of ginseng saponins on a rat visceral hypersensitivity model. AB - The 5-hydroxytryptamine3A (5-HT3) receptor is closely related with irritable bowel syndrome (IBS) in enteric nervous systems. We previously demonstrated that ginseng total saponins (GTS, also called ginsenosides), the active ingredients of Panax ginseng, inhibit the activity of 5-HT3A receptor channels expressed in Xenopus laevis oocytes. Here, we further investigated whether the in vitro inhibitory effect of ginsenosides on 5-HT3A receptor channel activity is coupled to in vivo attenuation of IBS. A rat model of IBS was induced by colorectal distention (CRD) and intracolonic infusion of 0.6% acetic acid (CRD-acetic acid), and visceral hypersensitivity was assessed by counting the contractions in the external oblique muscles of conscious rats during the 10 min distention period. We found that oral administration of GTS significantly and dose-dependently inhibited CRD-acetic acid-induced visceral hypersensitivity. The EC50 was 5.5+/ 4.7 mg/kg (95% confidence intervals: 1.2-15.7) and the inhibitory effect of GTS against visceral hypersensitivity persisted for 4 h. When we compared the effects of protopanaxadiol (PD) ginsenosides and protopanaxatriol (PT) ginsenosides against CRD-acetic acid-induced visceral hypersensitivity, we found that PT but not PD ginsenosides significantly attenuated the CRD-acetic acid-induced visceral hypersensitivity. These results indicate that PT ginsenosides of Panax ginseng might be the main active components for the attenuation of experimentally CRD acetic acid-induced visceral hypersensitivity, and may be clinically relevant for the future treatment of IBS. PMID- 16272702 TI - In vitro and in vivo antimicrobial action of tea: the commonest beverage of Asia. AB - The methanolic extract of leaves of Camellia sinensis (L) O. KUNTZE was screened for antimicrobial property against 111 bacteria comprising 2 genera of Gram positive and 7 genera of Gram negative bacteria. Most of these strains were inhibited by the compound at 10-50 microg/ml level and few strains were sensitive even at lower concentrations (5 microg/ml). The bacteria could be arranged in the decreasing order of sensitivity towards the compound in the following manner: Staphylococcus aureus, Vibrio cholerae, Escherichia coli, Shigella spp., Salmonella spp., Bacillus spp., Klebsiella spp. and Pseudomonas aeruginosa. The antibacterial activity of compound could also be confirmed in vivo. When it was given to Swiss strain of white mice at different dosages (30, 60 microg/mouse), it could significantly protect the animals challenged with 50 MLD of Salmonella typhimurium NCTC 74. According to Chi square test the in vivo data were highly significant (p<0.001). PMID- 16272704 TI - Presence of a motif conserved between Helicobacter pylori TNF-alpha inducing protein (Tipalpha) and penicillin-binding proteins. AB - Here we report a primary structure conserved between Helicobacter pylori (H. pylori)-tumor necrosis factor-alpha inducing protein (Tipalpha) and bacterial penicillin-binding proteins. H. pylori is a Gram-negative bacterium which plays a key part in carcinogenesis in the human stomach. We previously reported that Tipalpha has a carcinogenic potential as tumor promoter, and that it has no obvious homologue in other species. To investigate the structure-function relationship of Tipalpha and to predict its ancestral protein, we searched among proteins which have weak homology to Tipalpha in their primary structures, using Psi-Blast, and we identified numerous Gram-positive bacterial penicillin-binding proteins as weakly homologous to Tipalpha. Among these, several unique amino acids are conserved and form a motif-like structure. Phylogenic tree analysis indicated that Tipalpha is closer to the penicillin-binding proteins of Gram positive bacteria, based on their primary structures, than to H. pylori. This finding suggests that Tipalpha and penicillin-binding proteins are derived from a common ancestral protein, and that Tipalpha gene may be transferred horizontally from Gram-positive bacteria to H. pylori. PMID- 16272703 TI - Involvement of mast cells in the development of fibrosis in rats with postmyocarditis dilated cardiomyopathy. AB - Dilated cardiomyopathy (DCM) is a major cause of morbidity and mortality. Occurrence of myocardial fibrosis is an important event in the ventricular remodeling process, which takes place during DCM. Mast cells are well known inflammatory cells implicated in various biological phenomena. The involvement of mast cells in the development of myocardial fibrosis of DCM in rats after autoimmune myocarditis remains unknown. Nine-week-old male Lewis rats were immunized with cardiac myosin and divided into vehicle treated (group V) and disodium cromoglycate (DSCG), a mast cell stabilizer (24 mg/kg i.p.) treated (group DSCG) groups. The animals were sacrificed after 60 d of immunization. The myocardium was excised and preserved for histopathology and protein analysis. Myocardial levels of transforming growth factor (TGF) beta1 and collagen-III were quantified. Staining of mast cells was performed by toluidine blue. A significant correlation was obtained between myocardial fibrosis and cardiac mast cell density. DSCG reduced myocardial fibrosis besides preventing infiltration and degranulation of mast cells. Our findings confirm the active participation of mast cells in the progression of myocardial fibrosis in rats with postmyocarditis DCM. PMID- 16272705 TI - Piperazine propanol derivative as a novel antifungal targeting 1,3-beta-D-glucan synthase. AB - 1,3-beta-D-Glucan synthase, which synthesizes a main component of fungal cell wall, is one of the promising targets for antifungal agents. In order to identify novel chemical classes of 1,3-beta-D-glucan synthase inhibitors, we screened a chemical library monitoring inhibition of the Candida albicans 1,3-beta-D-glucan synthase activity. The piperazine propanol derivative GSI578 [(2,6-difluoro phenyl)-carbamic acid 3-(4-benzothiazol-2-yl-piperazine-1-yl)-propyl ester] was identified as a potent inhibitor against 1,3-beta-D-glucan synthase with an IC50 value of 0.16 microM. GSI578 exhibited in vitro antifungal activity against pathogenic fungi including C. albicans and Aspergillus fumigatus. Temperature sensitive mutations of the FKS1 gene in the Deltafks2 background of Saccharomyces cerevisiae, where FKS1 and FKS2 encode putative catalytic subunits of 1,3-beta-D glucan synthase, altered sensitivity to GSI578. This suggests that the antifungal activity of the piperazine propanol derivative has an effect on 1,3-beta-D-glucan synthase inhibition. Results of our initial evaluation suggest that the piperazine propanol derivative is a novel chemical structure of the class of antifungals which inhibit fungal cell growth by inhibiting fungal 1,3-beta-D glucan synthase. PMID- 16272706 TI - Synergistic effect of beta-cryptoxanthin and zinc sulfate on the bone component in rat femoral tissues in vitro: the unique anabolic effect with zinc. AB - The effect of the combination of beta-cryptoxanthin and zinc sulfate (zinc) on bone components in the femoral-diaphyseal and -metaphyseal tissues of young rats in vitro was investigated. Bone tissues were cultured for 48 h in a serum-free Dulbecco's modified Eagle's medium containing either vehicle, beta-cryptoxanthin (10(-9)-10(-7) M) or zinc sulfate (10(-6)-10(-4) M). The presence of beta cryptoxanthin (10(-9) M) or zinc (10(-6) M) did not have a significant effect on calcium content in the femoral-diaphyseal or -metaphyseal tissues. However, culture which combined beta-cryptoxanthin (10(-9) M) and zinc (10(-6) M) caused a significant increase in calcium content in the femoral-diaphyseal and metaphyseal tissues. Such an effect was not observed by the combination of beta cryptoxanthin (10(-9) M) plus genistein (10(-6) M) or menaquinone-7 (10(-6) M), or zinc (10(-6) M) plus genistein (10(-6) M) or menaquinone-7 (10(-6) M). Also, the combination of beta-cryptoxanthin (10(-9) M) plus zinc (10(-6) M) caused a remarkable increase in alkaline phosphatase activity and deoxyribonucleic acid (DNA) in the femoral-diaphyseal and -metaphyseal tissues, while their application alone did not have an effect on the enzyme activity or DNA content in the femoral tissues. The effect of the combination of beta-cryptoxanthin (10(-9) M) plus zinc (10(-6) M) in increasing calcium content, alkaline phosphatase activity, and DNA content in the femoral-diaphyseal and -metaphyseal tissues was completely prevented in the presence of cycloheximide (10(-6) M), an inhibitor of protein synthesis, or 5,6-dichloro-1-beta-D-ribofuranosylbenzimidazole (DBR), an inhibitor of transcriptional activity. This study demonstrates that the combination of beta-cryptoxanthin and zinc at a lower concentration has a synergistic effect on bone components in vitro. PMID- 16272707 TI - Comparison of the effects of cytoprotective drugs on human plasma adrenocorticotropic hormone and cortisol levels with continual stress exposure. AB - Cetraxate hydrochloride (cetraxate), ecabet sodium (ecabet), and sulpiride, which are cytoprotective drugs, have been used to treat peptic ulcers and acute or chronic gastritis. They are reported to improve mucosal blood flow in the stomach. One of the most important factors believed to cause gastric ulcers is mental and/or physiological stress. When people feel stress, the hypothalamo pituitary-adrenal (HPA) axis is activated. Therefore, corticotropin-releasing hormone (CRH), adrenocorticotropic hormone (ACTH), and cortisol can be indicators of stress. We examined the effects of cetraxate, ecabet and sulpiride on the plasma levels of ACTH and cortisol under stress conditions by repetitive blood sampling. Venous blood samples were taken before and 20-240 min after a single administration of the drugs or a placebo. A single dose of ecabet caused significant suppression of increases in plasma ACTH-like immunoreactive substance (IS) levels at 90 to 120 min and cortisol levels at 240 min, compared with the response to placebo. Sulpiride only suppressed increases in plasma cortisol levels at 180 to 240 min, compared with the response to placebo. A single dose of cetraxate had no effect on plasma ACTH-IS and cortisol levels. Ecabet may have a modulatory effect on the HPA axis while sulpiride may have a partial modulatory effect on the HPA axis. These effects might be beneficial in stress-related disease. PMID- 16272708 TI - Enzymatic measurement of tryptase-like protease release from isolated perfused guinea pig heart during ischemia-reperfusion. AB - To elucidate the details of tryptase release from the heart during ischemia reperfusion (I/R), we attempted the enzymatic measurement of tryptase release from the isolated guinea pig heart perfused by the Langendorff mode I/R model. Tryptase-like activity in the effluent was monitored by the hydrolysis of L-Pyr Gly-Arg-MCA. Tryptase-like protease and histamine were rapidly released from heart during ischemia within 10 min. After reperfusion, tryptase-like protease levels decreased, achieving stabilization. The tryptase-like protease activity in the effluent was inactivated by serine protease inhibitors. The pattern of inhibition was similar to those of guinea pig and human lung tryptase. In conclusion, tryptase was released into the coronary effluent during ischemia, but not during reperfusion in guinea pig heart. PMID- 16272709 TI - Antidiabetic effect of Nitobegiku, the herb Tithonia diversifolia, in KK-Ay diabetic mice. AB - Nitobegiku (the herb of Tithonia diversifolia (HEMSL) A. GRAY) has been used as a medicinal plant for diabetes. The antidiabetic effect of an 80% ethanol extract of Nitobegiku (Td) was investigated in KK-Ay-mice, an animal model of type 2 diabetes. Td (500 mg/kg body weight) reduced the blood glucose of KK-Ay mice 7 h after a single oral dose. No change in blood glucose in Td-treated normal mice (ddY) was seen. Td (500 mg/kg) reduced blood glucose in KK-Ay mice 3 weeks after a single oral dose and also significantly lowered plasma insulin in KK-Ay mice under similar conditions. Td-treated KK-Ay mouse blood glucose was significantly decreased in an insulin tolerance test. These results support the hypothesis that Td improves glucose metabolism by reducing insulin resistance. Therefore, Nitobegiku may be useful for the treatment of type 2 diabetes. PMID- 16272710 TI - Phenolic constituents of galla Rhois with hepatoprotective effects on tacrine- and nitrofurantoin-induced cytotoxicity in Hep G2 cells. AB - The bioassay-guided fractionation of the MeOH extract of Galla Rhois furnished two hepatoprotective compounds, an equilibrium mixture of 3-galloyl-gallic acid and 4-galloyl-gallic acid isomers (3), 1,2,3,4,6-penta-O-galloyl-beta-D-glucose (4), and two inactive phenolic compounds, gallic acid methyl ester (1) and gallic acid (2). Compounds 3 and 4 showed significant hepatoprotective effects with EC50 values of 70.39+/-5.4 and 29.51+/-0.7 microM, respectively, against tacrine induced cytotoxicity, and 150.9+/-6.4 and 23.81+/-0.5 microM, respectively, against nitrofurantoin-induced cytotoxicity in Hep G2 cells. PMID- 16272711 TI - Antimutagenicity and cytotoxicity of the constituents from the aerial parts of Rumex acetosa. AB - Four anthraquinones isolated for the first time from the aerial parts of Rumex acetosa (Polygonaceae), a Korean and a Japanese medicinal plant, and two synthetic derivatives were examined for their cytotoxicities against five cultured human tumor cell lines, i.e. A549 (non-small cell lung), SK-OV-3 (ovary), SK-MEL-2 (melanoma), XF498 (central nerve system) and HCY15 (colon), using the Sulfrhodamine-B method in vitro and antimutagenic activities by Ames test with Salmonella typhimurium TA98 and TA100 and SOS chromotest with E. coli PQ37. Among the tested compounds, emodin strongly inhibited the proliferation of each examined tumor cell line with IC50 values ranged from 2.94 to 3.64 microg/ml and showed potent antimutagenic activities with 71.5% and 53.3% at the concentration of 0.1 mg/plate against the mutagens, NPD and sodium azide, respectively. Its antigenotoxic activity was also very effective at the final concentration of 10 microg/reaction tube against the mutagens, MNNG and NQO by SOS chromotest, reducing the induction factors by 19.6% and 43.5%, respectively. The structure-activity correlation study suggests that an additional OH group at C-6 position in the anthraquinone nucleus may play an important role for their cytotoxicities and an introduction of OH- or OCH3 group at C-6 position is necessary for their antimutagenicities. PMID- 16272712 TI - Bofutsushosan, a traditional Chinese formulation, prevents intimal thickening and vascular smooth muscle cell proliferation induced by balloon endothelial denudation in rats. AB - Bofutsushosan (BOF), a traditional Chinese formulation (Kampo formulation in Japanese), is widely used for patients with obesity and hyperlipidemia resulting from long-term inappropriate lifestyles. Since atherosclerosis, a lifestyle related disease, is accompanied by an abnormal accumulation of vascular smooth muscle cells (VSMCs) in the intimal area of the artery, we investigated the preventive effect of BOF on intimal thickening. Oral administration of BOF extracts 3 d before and 7 d after balloon endothelial denudation dose dependently suppressed the intimal thickening and proliferation of VSMCs in the intimal area in rat carotid arteries. This model has a similar pathologic process to atherosclerosis and is considered to be an "accelerated atherosclerosis" model. BOF extract also dose dependently inhibited the migration of cultured VSMCs. BOF extract suppressed serum lipid levels, which are a major risk factor for atherosclerosis. These findings clarified the usefulness of BOF in cardiovascular risk-reduction therapy. PMID- 16272713 TI - Effect of solution composition of plasmid DNA on gene transfection following liver surface administration in mice. AB - We investigated the effect of plasmid DNA (pDNA) solution composition on gene transfection following liver surface administration in mice. Gene transfection experiments in situ and in vivo were performed using the following pDNA solutions: dextrose solution, NaCl solution, phosphate buffer, phosphate-buffered saline, Tris/HCl buffer with EDTA, Tris/HCl buffer with EDTA and Triton X-100, and water. In in situ experiments, we used a glass cylindrical diffusion cell that limited the contact area between the liver surface and the naked pDNA solution. The gene transfection at the site of diffusion cell attachment increased in hypotonic solution, and decreased in hypertonic solution, compared with isotonic solution. In in vivo experiments, instillation of naked pDNA solution onto the liver surface using a micropipette caused no significant differences in gene transfection in the applied lobe. These results suggest that it is important to select the optimal pDNA solution composition to control the gene transfection. PMID- 16272714 TI - Quantitative evaluation of effect of renal failure on the pharmacokinetics of panipenem in rats. AB - The pharmacokinetics of panipenem in experimental renal failure animal models was investigated in order to identify the appropriate covariates affecting the pharmacokinetic behavior. Panipenem and betamipron were administered intravenously to rats with a variety of renal failures, such as nephritis induced by glycerol, gentamicin, uranium and antiserum against glomerular basement membrane as well as 5/6 subtotal nephrectomy. Panipenem in plasma and urine was determined and pharmacokinetic analysis was performed using a one-compartment open model. The elimination half-life prolonged and total body clearance, renal clearance (CL(R)) and renal excretion ratio were decreased according to the renal function, i.e. control>glycerol>anti-GBM=gentamicin>nephrectomy=uranium in order. However, distribution volume was consistent in all models. CL(R) showed strong positive correlation with the glomerular filtration rate in spite of a weak correlation with the reciprocal of blood urea nitrogen. However, no obvious correlation was observed with secretory clearance of N-1-methylnicotinamide. This preliminary information based on animal model might be useful for designing pharmacokinetic studies in special population at early stage of new drug development. PMID- 16272715 TI - Regulation effect of 2',4',7-trihydroxyisoflavone on the expression of matrix metalloproteinase-1, 2 in ultraviolet-B irradiated primary cultured old aged human skin fibroblasts. AB - Long term and repeated exposure of UV light on the skin often induces chronic skin diseases such as skin cancer as well as photoaging, and the mechanisms of these skin damages are closely associated with up-regulation of matrix metalloproteinase's (MMPs) activities. We investigated the effect of 2',4',7 trihydroxyisoflavone purified from the whole plants of Viola hondoensis W. BECKER et H BOISSIEU (Violaceae) on the expression of MMPs in UV-B irradiated old aged human skin fibroblasts. 2',4',7-trihydroxyisoflavone markedly reduced UV-induced MMP-1 expression, but not MMP-2, at the both mRNA and protein levels in a dose dependent manner. Our report is the first description for the ability of 2',4',7 trihydroxyisoflavone to regulate MMP-1 expression from ultraviolet-B irradiated primary cultured old aged human skin fibroblasts. PMID- 16272716 TI - Meso-dihydroguaiaretic acid from Machilus thunbergii SIEB et ZUCC., and its effects on the expression of matrix metalloproteinase-2, 9 cause by ultraviolet irradiated cultured human keratinocyte cells (HaCaT). AB - Ethanol and aqueous extracts of Machilus thunbergii used traditionally for the treatments a wide variety of diseases were screened in vitro for the matrix metalloproteinase (MMP)-9 inhibitor actions. Meso-dihydroguaiaretic acid from the stems bark of Machilus thunbergii showed significant MMP-9 inhibition in human keratinocyte cells cause by ultraviolet irradiation. Here we investigated the effect of meso-dihydroguaiaretic acid, which was isolated from Machilus thunbergii, on UV-induced premature skin aging. We studied the effect of meso dihydroguaiaretic acid on UV-induced MMP-9 expression in an immortalized human keratinocyte cell line, HaCaT, in vitro. Acute UV irradiation induced MMP-9 expression at both the mRNA and protein levels and meso-dihydroguaiaretic acid suppressed this UV-induced MMP-9 expression in a dose-dependent manner. Taken together, these results show that meso-dihydroguaiaretic acid can prevent the harmful effects of UV that lead to skin aging. Therefore, we suggest that meso dihydroguaiaretic acid should be viewed as a potential therapeutic agent for preventing and/or treating premature skin aging. PMID- 16272717 TI - Development of new synthetic methods and its application to total synthesis of nitrogen-containing bioactive natural products. AB - A group of naturally occurring substances containing nitrogen is widely distributed in plants as well as in fungi, animal, marine organisms, and insects, and many exhibit significant biological activity. These natural products with a huge variety of chemical structures include antibiotics, antitumor agents, immunostimulants, drugs affecting the cardiovascular and central nervous systems, analgesics etc. The diverse activities and low natural abundance of this group of natural products when coupled with their molecular complexity warrant development of new and efficient synthetic methods and strategy for the total synthesis of these products, in particular alkaloids. The purpose of this review is to describe some of our achievements in the total synthesis of the naturally occurring bases including the Dendrobatid alkaloids pumiliotoxin B and allopumiliotoxin A, the anitibiotic streptazolin, the tricyclic marine alkaloids isolated from the ascidians such as fasicularin, lepadiformine, and cylindricine C, and the dimeric monoterpene alkaloid incarvillateine as well as the formal total synthesis of the spirocyclic marine alkaloids halichlorine and pinnaic acid, which are isolated from the Japanese marine sponge and the Okinawan bivalve, respectively. PMID- 16272718 TI - Semisynthesis and antitumor activities of new styryl-lactone derivatives. AB - Nineteen new derivatives of the naturally occurring compound, goniothalamin, were prepared by chemical modification and semi-synthetic methods. The antitumor activities of these derivatives and goniothalamin were evaluated in vitro against human tumor cell lines, and most of them showed an inhibitory effect against HL 60 cancer cells. The derivatives 10-nitro-goniothalamin and 10-amino goniothalamin gave selective inhibition concentration (IC50) of 1.10 and 1.14 microg/ml, respectively, against human stomach cancer SGC-7901 cells, while that of etoposide (vp-16) as the positive control was 6.07 microg/ml. Finally, the partition coefficients, logP (pi values), of these derivative molecules, were evaluated by calculating the additive approximate organic fragment logP value. PMID- 16272719 TI - Simultaneous determination of major bioactive components in Qingkailing injection by high-performance liquid chromatography with evaporative light scattering detection. AB - High-performance liquid chromatography with evaporative light scattering detection (HPLC/ELSD) was established for simultaneous determination of seven major bioactive components of Qingkailing injection including adenosine, geniposide, chlorogenic acid, baicalin, ursodeoxycholic acid, cholic acid, and hyodeoxycholic acid. The proposed method was applied to analyze ten various Qingkailing injections and produced data with acceptable linearity, repeatability, precision and accuracy having a limit of detection (LOD) of 10-50 ng. In comparison with UV detection, HPLC/ELSD permits the determination of non chromophoric compounds without prior derivatization, and shows good compatibility to the multi-components of complex analytes. The proposed method is a useful alternative for routine analysis in the quality control of traditional Chinese medicine. PMID- 16272720 TI - Intercalation compound of diclofenac sodium with layered inorganic compounds as a new drug material. AB - The intercalation reaction of diclofenac sodium (DFS) with layered inorganic compounds, gamma-titanium phosphate (gamma-TiP), proton type titanium oxide (H TiO2) and sodium type synthetic mica (Na-TSM), was examined on. The direct reaction of DFS in ethanol-water mixed solvent resulted in the large amount accommodation of DFS. The amount of intercalated DFS was the order of gamma-TiP>H TiO2>Na-TSM corresponding to the order of acidity. The intercalation using phospholiopids was also examined to assist the intercalation reaction. However, the amount of intercalated DFS was rather small in comparison with those in the direct reaction. DFS accommodated in gamma-TiP dissolved into neutral and basic buffer solution stoichiometry while scarcely dissolved in the acidic solution. The mechanism of the intercalation and reverse dissolution was successfully accounted according to the ion-exchange mechanism between Na+ in DFS and H+ in gamma-TiP. The dissolution from tablet of DFS/gamma-TiP intercalation compound was examined by using a disintegrator. It was found that the dissolution rate appropriately controlled by mixing the disintegrator. The present results suggested the different possibilities in the clinical field to use layered inorganic compounds such as drug delivery system (DDS). PMID- 16272721 TI - Synthesis of selenium-containing polyphenolic acid esters and evaluation of their effects on antioxidation and 5-lipoxygenase inhibition. AB - Six novel selenium-containing polyphenolic acid esters were synthesized and evaluated as antioxidants and 5-lipoxygenase inhibitors. Synthesis of the title compounds involved the Mitsunobu reaction of polyphenolic acids with 2 phenylselenoethanol. Compounds and were found to be very effective antioxidants and 5-lipoxygenase inhibitors with activity comparable to or better than caffeic acid (3,4-dihydroxycinnamic acid) phenethyl ester (CAPE). PMID- 16272722 TI - Hypoglycemic activity of a new carbohydrate isolated from the roots of Psacalium peltatum. AB - A new ulopyranose isolated from aqueous extract of roots and rhizomas of Psacalium peltatum has been determined to have hypoglycemic activity at doses of 100 mg/kg, comparable to that of tolbutamide and insulin in alloxan diabetic mice. The skeletal structure of the new compound was established by spectral analysis. PMID- 16272723 TI - Quantitative measurements of oxidative stress in mouse skin induced by X-ray irradiation. AB - To find efficient methods to evaluate oxidative stress in mouse skin caused by X ray irradiation, several markers and methodologies were examined. Hairless mice were irradiated with 50 Gy X-rays and skin homogenates or skin strips were prepared. Lipid peroxidation was measured using the skin homogenate as the level of thiobarbituric acid reactive substances. The level of lipid peroxidation increased with time after irradiation and was twice that of the control at 78 h. ESR spectra of skin strips showed a clear signal for the ascorbyl radical, which increased with time after irradiation in a manner similar to that of lipid peroxidation. To measure levels of glutathione (GSH) and its oxidized forms (GSSG) simultaneously, two HPLC methods, sample derivatization with 1-fluoro-2,4 dinitrobenzene and detection with a UV detector (method A) and no derivatization and detection with an electrochemical detector (method B), were compared and the latter was found to be better. No significant change was observed within 24 h after irradiation in the levels of GSH and GSSG measured by method B. The GSH/GSSG ratio may be a less sensitive parameter for the evaluation of acute oxidative stress caused by X-ray irradiation in the skin. Monitoring the ascorbyl radical seems to be a good way to evaluate oxidative stress in skin in vivo. PMID- 16272725 TI - Biotransformation of citrus aromatics nootkatone and valencene by microorganisms. AB - Biotransformations of the sesquiterpene ketone nootkatone from the crude drug Alpiniae Fructus and grapefruit oil, and the sesquiterpene hydrocarbon valencene from Valencia orange oil were carried out with microorganisms such as Aspergillus niger, Botryosphaeria dothidea, and Fusarium culmorum to afford structurally interesting metabolites. Their stereostructures were established by a combination of high-resolution NMR spectral and X-ray crystallographic analysis and chemical reaction. Metabolic pathways of compounds and by A. niger are proposed. PMID- 16272724 TI - Bioactive constituents from Chinese natural medicines. XV. Inhibitory effect on aldose reductase and structures of Saussureosides A and B from Saussurea medusa. AB - The 80% aqueous acetone extract from the whole plant of Saussurea medusa MAXIM. was found to inhibit rat lens aldose reductase (IC50=1.4 microg/ml). From this extract, flavonoids, lignans, and quinic acid derivatives were isolated together with two new ionone glycosides, saussureosides A and B. Their absolute stereostructures were elucidated on the basis of chemical and physicochemical evidence including the application of modified Mosher's method. In addition, some isolates were found to show an inhibitory effect on aldose reductase. PMID- 16272726 TI - Preparation of new nitrogen-bridged heterocycles. 58. Syntheses and intramolecular arene-pi interactions of 3-(allylthio)- and 3 (propargylthio)thieno[3,4-b]indolizine derivatives. AB - Various thieno indolizine derivatives having an allylthio or propargylthio group at the 3-position were prepared and their intramolecular arene-pi interactions were investigated. Their 1H-NMR spectra showed significant low-field shifts (delta 0.10-0.34 ppm) to the 5-proton on the thieno indolizine ring, and this effect was the reverse to that observed in 3-(arylmethylthio)thieno indolizines. However, their UV spectra exhibited a characteristic absorption band due to the arene-pi interaction near 430 nm and these values were almost similar to those for arene-arene interaction of 3-arylmethylthio derivatives though their molar extinction coefficients were largely varied by the 3-substituents. Furthermore, both types of gauche conformations in which the intramolecular arene-pi interactions are possible in one form and impossible in the other were confirmed by X-ray analyses of some compounds. PMID- 16272727 TI - Superoxide anion-scavenging effect of 2-amino-1,3-selenazoles. AB - We investigated the superoxide anion scavenging effects of thirteen 2-amino-1,3 selenazoles using a highly sensitive quantitative chemiluminescence method. At 166 microM, the 2-amino-1,3-selenazoles scavenged in the range of 14.3 to 96.7% of O2-. 2-Piperidino-1,3-selenazole and 4-phenyl-2-piperidino-1,3-selenazole exhibited the strongest superoxide anion-scavenging activity among the 2-amino 1,3-selenazoles. The 50% inhibitory concentrations (IC50) of 2-piperidino-1,3 selenazole and 4-phenyl-2-piperidino-1,3-selenazole were determined to be 4.03 microM and 92.6 microM, respectively. Thus, these compounds acted in vitro as effective O2- scavengers. PMID- 16272729 TI - A rapid and reliable solid-phase extraction method for high-performance liquid chromatographic analysis of opium alkaloids from papaver plants. AB - A rapid and reliable solid-phase extraction method for HPLC analysis of opium alkaloids from Papaver plants was established. Fifty mg of dried and powdered plant sample was extracted with 5 ml of 5% acetic acid for 30 min under sonication. After centrifugation, 3 ml of the supernatant was loaded on a reversed-phase cation-exchange solid-phase extraction cartridge. After seriate washings with 0.1 M hydrochloric acid and methanol, alkaloids were eluted with a mixture of 28% ammonia and methanol (1:19). The eluate was concentrated under nitrogen stream at 40 degrees C and the residue was dissolved in 50% aqueous methanol for high performance liquid chromatographic analysis. With this solid phase extraction method, the recovery of morphine, codeine, oripavine, thebaine, papaverine, noscapine and sanguinarine was from 99.94 to 112.18% when the standard alkaloids were added to the plant samples. Opium alkaloids of a variety of genus Papaver plants cultivated in a field and phytotron were analyzed by this method. PMID- 16272728 TI - Comparison of the glass transition temperature and fragility parameter of isomalto-olygomer predicted by molecular dynamics simulations with those measured by differential scanning calorimetry. AB - The purpose of this study is to examine whether molecular dynamics (MD) simulations using a commercially available software for personal computers can estimate the glass transition temperature (Tg) of amorphous systems containing pharmaceutically-relevant excipients. MD simulations were carried out with an amorphous matrix model constructed from isomaltoheptaose, and the Tg estimated from the calculated density versus temperature profile was compared with the Tg measured by differential scanning calorimetry (DSC) for freeze-dried isomalto oligomer having an average molecular weight close to that of isomaltoheptaose. The Tg values determined by DSC were lower by 10 to 20 K than those extrapolated from the Tg values estimated by MD simulation. Fragility parameter was estimated to be 56 and 51 from MD simulation and from DSC measurement, respectively. Thus, the results suggest that MD simulation can provide approximate estimates for the Tg and fragility parameter of amorphous formulations. However, a reduction of the cooling rate, achievable by sufficiently elongating the simulation duration, is necessary for more accurate estimation. PMID- 16272730 TI - Antitumor activity of Pulsatilla koreana saponins and their structure-activity relationship. AB - Seventeen saponins isolated from the root of Pulsatilla koreana were examined for their in vitro cytotoxic activity against the human solid cancer cell lines, A 549, SK-OV-3, SK-MEL-2, and HCT15, using the SRB assay method, and their in vivo antitumor activity using BDF1 mice bearing Lewis lung carcinoma (LLC). The saponins 5-17, with a free acidic functional group at C-28 of aglycon, exhibited moderate to considerable cytotoxic activity, however, the saponins 1-4, esterified with a trisaccharide at C-28 of aglycon, did not exhibit cytotoxic activity (ED50; >300 microM). Among them, oleanolic acid 3-O-alpha-L rhamnopyranosyl-(1-->2)-[beta-D-glucopyranosyl-(1-->4)]-alpha-L-arabinopyranoside (10) exhibited the most potent cytotoxic activity (ED50; 2.56, 2.31, 1.57, 8.36 microM, respectively). In vivo test, hederagenin 3-O-alpha-L-rhamnopyranosyl-(1- >2)-[beta-D-glucopyranosyl-(1-->4)]-alpha-L-arabinopyranoside (6, Inhibition Ratio, IR; 66.9%) exhibited more potent antitumor activity than taxol (IR; 35.8%) and doxorubicin (IR; 62.1%). Also, hedragenin 3-O-beta-D-glucopyranosyl-(1-->4)-O beta-D-glucopyranosyl-(1-->3)-O-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L arabinopyranoside (17, IR; 50.3%) exhibited potent antitumor activity. These two saponins were identically comprised of a hederagenin aglycon moiety and a sugar sequence O-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranoside at C-3 of the hederagenin, suggesting that the two elements are essential factors for the antitumor activity. PMID- 16272732 TI - A one-step synthesis of 2-Alkyl-5-hydroxychromones and 3-Alkoyl-2-alkyl-5 hydroxychromones. AB - 2-Alkyl-5-hydroxychromones (2-alkyl-5-hydroxy-4-oxo-4H-1-benzopyran) and 3-alkoyl 2-alkyl-5-hydroxychromones (3-alkoyl-2-alkyl-5-hydroxy-4-oxo-4H-1-benzopyran) were prepared in one-step and one pot reaction by condensation of 2',6' dihydroxyacetophenone with an alkoyl chloride in the presence of K2CO3. PMID- 16272731 TI - Microbial hydroxylation of pregnenolone derivatives. AB - Pregnenolone and pregnenolone acetate were incubated with the fungi Cunninghamella elegans, Rhizopus stolonifer and Gibberella fujikuroi. Incubation of with C. elegans yielded metabolites, 3beta,7beta,11alpha-trihydroxypreg-5-en 20-one, 3beta,6alpha,11alpha,12beta,15beta-pentahydroxypreg-4-en-20-one and 3beta,6beta,11alpha-trihydroxypreg-4-en-20-one, while incubation with G. fujikuroi yielded two known metabolites, 3beta,7beta-dihydroxypregn-5-en-20-one and 6beta,15beta-dihydroxypreg-4-ene-3,20-dione. Metabolites and were found to be new. Fermentation of by C. elegans yielded four known oxidative metabolites, androsta-1,4-diene-3,17-dione, 6beta,15beta-dihydroxyandrost-4-ene-3,17-dione and 11alpha,15beta-dihydroxypreg-4-ene-3,20-dione. Fermentation of with R. stolonifer yielded two known metabolites, 11alpha-hydroxypreg-4-ene-3,20-dione and. Compounds were screened for their cholinesterase inhibitory activity in a mechanism-based assay. PMID- 16272733 TI - Formosadimers A, B, and C from the Bark of Calocedrus macrolepis var. formosana. AB - Formosadimers A, B, and C, together with one known compound, sugikurojin B, have been isolated from the bark of Calocedrus macrolepis var. formosana. Formosadimers A, B, and C, and sugikurojin B are dimers of the abietane-O abietane type. Their structures were elucidated principally based on spectroscopic data. PMID- 16272734 TI - Cyclooxygenase-2 inhibitory phenylbutenoids from the rhizomes of Zingiber cassumunar. AB - Phenylbutenoids isolated previously from the CHCl3 extracts of the rhizomes of Zingiber cassumunar, were evaluated for their cyclooxygenase-2 (COX-2) inhibitory activity along with a new isolate, from the n-BuOH extracts of this plant. The COX-2 inhibitory assay was performed by measuring prostaglandin E2 production in lipopolysaccharide-stimulated mouse macrophage RAW 264.7 cells. Two phenylbutenoid dimers, and, exhibited considerable activity with IC50 values of 2.71 and 3.64 microM. Two phenylbutenoid monomers, and, showed moderate activity (IC50 14.97, 20.68 microM, respectively). The other three phenylbutenoids, were found to be inactive. Compound was elucidated as a new phenylbutenoid glycoside, namely, (E)-4-(3,4-dimethoxyphenyl)but-3-en-1-O-beta-D-glucopyranoside by spectral analysis including various 1D- and 2D-NMR experiments. PMID- 16272735 TI - Phenyl polypropanoids from Lindelofia stylosa. AB - A phytochemical investigation on the aerial parts of Lindelofia stylosa has resulted in the isolation of seven phenyl propanoids. This includes three analogs of lithospermic acid, along with rosmarinic acid and its ester derivatives. Compound 1 was identified as a new natural product. These compounds were studied for their antioxidant properties. PMID- 16272736 TI - Quantitative determination of incarvillateine in Incarvillea sinensis by solid phase extraction and high performance liquid chromatography. AB - Method for rapid quantitative analysis of incarvillateine in Incarvillea sinensis by high-performance liquid chromatography (HPLC) has been developed. The sample preparation involves solid phase extraction (SPE) with a mixed-mode reversed phase and cation-exchange cartridge. The linear calibration range for incarvillateine was 0.002-0.5 mg/ml. The limit of detection was 0.35 microg/ml (S/N=3). Intra- and interday precisions were less than 0.36% (n=6) and 1.61% (n=18), respectively. The recovery of incarvillateine was 97.61-102.44% with the relative standard deviation (RSD) ranging from 0.63 to 1.93% (n=3). This method was proposed as a simple, rapid and accurate method for quantitative determination of incarvillateine content in various samples of Incarvillea sinensis collected from different areas of China. PMID- 16272737 TI - New bis-spirolabdane-type diterpenoids from Leonurus heterophyllus Sw. AB - Twelve natural bis-spirolabdane-type diterpenoids, including eight new, named leoheteronones A-E, 15-epileoheteronones B, D, and E, and four known leopersin B, 15-epileopersin B, leopersin C, and 15-epileopersin C, together with hispanone and galeopsin were isolated from the aerial parts of the medicinal plant Leonurus heterophyllus SW. (Lamiaceae) grown in Vietnam. Their structures were determined by spectroscopic analyses. The current study emphasized the accumulation of C-15 oxygenated bis-spirolabdane-type diterpenoids of both 13R and 13S configurations in L. heterophyllus. PMID- 16272738 TI - Identification and comparative determination of senkyunolide A in traditional Chinese medicinal plants Ligusticum chuanxiong and Angelica sinensis by HPLC coupled with DAD and ESI-MS. AB - Using the HPLC/DAD/ESI/MS method, the qualitative and quantitative analysis of senkyunolide A (SA) in the rhizomes of Ligusticum chuanxiong (Rhizoma chuanxiong; CX) and roots of Angelica sinensis (DG) was established. As a result, it was found that SA is a characteristic standard compound for the quality evaluation and chemical differentiation between CX and DG. Methanol was chosen in the preparation of standard solutions and extraction of samples based on the stability data. The identity of SA in CX and DG was unambiguously determined based on the quasimolecular ions in ESI-MS. A comprehensive validation of the method, including sensitivity, linearity, reproducibility and recovery, was conducted using the optimized chromatographic conditions. The linear calibration curve was acquired with R2>0.999 and limit of detection (S/N=3) was estimated to be 12.5 mug/g. The reproducibility was evaluated by repeated sample injection and replicated analysis of samples with the relative standard deviation (RSD) value found within 0.68%. The recovery rates of SA varied within the range of 96.91 101.50% with RSD less than 2.38%. In the present work, the contents of SA were quantified within 3.94-9.14 mg/g and 0.108-0.588 mg/g for 12 batches each of CX and DG. The results demonstrated that SA is a useful standard compound for the quality evaluation and chemical differentiation between CX and DG. The analytical procedure is precise and reproducible and thus suitable for the analysis of a large number of samples. PMID- 16272739 TI - Establishment of GC-MS fingerprint of fresh Houttuynia cordata. AB - Fresh Houttuynia cordata THUNB. is a Chinese materia medica generally used in Chinese medicine therapy. It possesses the actions of clearing heat, eliminating toxins, reducing swelling, discharging pus and relieving stagnation. However, dry H. cordata has traditionally been used in clinical application instead of the fresh counterpart. In this paper, the chemical profiles of H. cordata were established using fingerprinting techniques. A modified GC-MS method was developed in the comparison of fingerprints among fresh and dry herbs of H. cordata. It was shown that the varieties, as well as relative levels of chemical components, in the fresh herb were more abundant than in the dry counterpart. Fingerprinting profiles were found to be consistent for fresh herbs acquired from various production areas, but the relative abundance of peaks were varied. Besides, the chemical components among different medicinal portions of fresh herbs were found to be inconsistent. The developed fingerprint can be successfully applied to distinguish between fresh and dry herbs, as well as determining differentiation among different medicinal portions. PMID- 16272740 TI - Preparation of 4-pentenoic acid ester of Neu5Ac and 4-pentenyl glycoside of Neu5Ac and their application to glycosylation. AB - Novel sialosyl donors, 4-pentenoic acid ester of N-acetylneuraminic acids (Neu5Ac) and 4-pentenyl glycoside of Neu5Ac were successfully prepared from the corresponding per-O-acetylated 2-hydroxy and 2-chloro derivatives of Neu5Ac, respectively and applied to the synthesis of O-sialosides. PMID- 16272741 TI - Chromatographic separation of (-)-ephedrine and (+)-pseudoephedrine in the traditional Chinese medicinal preparation Jiketing granule. AB - (-)-Ephedrine (EP) and (+)-Pseudoephedrine (PEP) in Jiketing granule were separated by TLC, RP-HPLC, GC, HPLC-MS, and the chromatographic phenomenon was interpreted by the topological indices. The observed chromatographic separations of investigated components were compared. This comparison indicated that simply methods as C18 RP-HPLC and GC are the more effective techniques, and more suitable than others for this preparation, for the separation of two alkaloids. PMID- 16272742 TI - Individual and simultaneous spectrophotometric determination of dapsone and metoclopramide HCl in pharmaceutical dosage forms and synthetic binary mixtures. AB - A rapid, sensitive and selective spectrophotometric method has been developed for the quantitative determination of dapsone (DAP) and metoclopramide hydrochloride (MCP) in both pure and dosage forms. Individual and simultaneous methods are based on the diazo coupling reaction of these drugs with benzoylacetone (BAC) in alkaline medium. The resulting azo dyes exhibit maximum absorption at 437 and 411 nm with a molar absorptivity of 4.14x10(4) and 2.97x10(4) l mol-1 cm-1 for DAP and MCP, respectively. Simultaneous determination of DAP and MCP was developed utilizing first-order digital derivative spectrophotometry. All variables have been optimized. No interferences were observed from drug excipients and the validity of the methods was tested against reference methods. PMID- 16272743 TI - Highly regioselective palladium-catalyzed annulation reactions of heteroatom substituted allenes for synthesis of condensed heterocycles. AB - We have developed a highly regioselective synthesis of heterocycles via palladium catalyzed annulation reaction of heteroatom-substituted allenes. Various aryl halides were reacted and one regioisomer was observed exclusively in all reactions. In addition, subsequent functionalizations of annulated products were carried out using alkyl metal reagents, and the introduction of alkyl moieties was accomplished. PMID- 16272744 TI - Preparation of (Cyanomethylene)tributylphosphorane: a new mitsunobu-type reagent. AB - (Cyanomethylene)tributylphosphorane (CMBP), which promoted the alkylation of various nucleophiles (HA) with alcohols (ROH) to give RA (Mitsunobu-type reaction), was prepared in two steps starting from chloroacetonitrile. PMID- 16272745 TI - Post-biotinylation of photocrosslinking by Staudinger-Bertozzi ligation of preinstalled alkylazide tag. AB - Post-biotinylation of the alkyl azide derivative of trifluoromethyl phenyldiazirine (TPD) was elucidated to apply a photoaffinity biotinylation technique. A photo-modified polyvinilidene difluoride (PVDF) membrane was used as a photolabeled component and we introduced biotin by Staudinger-Bertozzi ligation. The 15 pmol amount of biotinylated reagent was still effective for the visualization of cross-linked product on the matrix. The results show the potential utility of alkyl azide carrying TPD derivatives in the application of photoaffinity biotinylation, which could be useful for the ligands with tight structural requirements. PMID- 16272746 TI - Highly efficient production of nootkatone, the grapefruit aroma from valencene, by biotransformation. AB - Nootkatone, the most important and expensive aromatic of grapefruit, decreases the somatic fat ratio, and thus its demand is increasing in the cosmetic and fiber sectors. A sesquiterpene hydrocarbon, (+)-valencene, which is cheaply obtained from Valencia orange, was biotransformed by the green algae Chlorella species and fungi such as Mucor species, Botryosphaeria dothidea, and Botryodiplodia theobromae to afford nootkatone in high yield. PMID- 16272747 TI - A fission yeast SNAP-25 homologue, SpSec9, is essential for cytokinesis and sporulation. AB - The soluble NSF attachment protein 25 (SNAP-25) is a component of the SNARE complex that is essential for regulated exocytosis in diverse cell types. Here, we identified a fission yeast SNAP-25 homologue, SpSec9. The sec9+ gene was essential for vegetative growth. sec9 mRNA was detected in vegetative cells and further increased during sporulation. This increase during sporulation was dependent on Mei4, a meiosis-specific transcription factor. A sporulation deficient sec9 mutant was isolated by random PCR mutagenesis (sec9-10). The sec9 10 mutant also exhibited temperature sensitivity for growth and cell division was found to arrest before completion of cell separation at restrictive temperatures. In sec9-10 cells, the forespore membrane was normally initiated near spindle pole bodies during meiosis II. However, subsequent extension of the membrane was severely impaired. These results indicate that SpSec9 plays an important role both in cytokinesis and in sporulation. PMID- 16272748 TI - A molecular functional study on the interactions of drugs with plasma proteins. AB - The binding of drugs to plasma proteins, such as albumin and alpha1-acid glycoprotein (AGP) is a major determinant in the disposition of drugs. A topology analysis of drug binding sites on HSA and AGP was determined using various methods, including spectroscopy, QSAR, photoaffinity labeling and site directed mutagenesis. Recombinant albumin was found to be useful for rapidly identifying drug binding sites. The binding sites on AGP are not completely separated but are partially overlapped, and Trp, Tyr, Lys and His residues in the drug binding pockets play important roles in this process. Drug displacement is somewhat complex, due to the involvement of multiple effects. The reduced binding in uremic patients may be explained by a mechanism that involves a combination of direct displacement by free fatty acids as well as cascade effects of free fatty acids and unbound uremic toxins for significant inhibition in serum binding. Albumin-containing dialysate is useful for the extracorporeal removal of endogenous toxins and in the treatment of drug overdoses. Oxidized albumin is a useful biomarker for the quantitative and qualitative evaluation of oxidative stress. Interestingly, AGP undergoes a structural transition to a unique structure that differs from the native and denatured states, when it interacts with membranes. PMID- 16272749 TI - Long-term levothyroxine treatment decreases the oral bioavailability of cyclosporin A by inducing P-glycoprotein in small intestine. AB - We have noticed that the trough level of blood concentration of cyclosporin A (CyA) tends to be lower in patients receiving long-term oral levothyroxine (LTX) than in patients not receiving LTX. We confirmed this clinical observation in experiments using Wistar rats orally given LTX (8 microg/kg) or saline (control) for 3 weeks, followed by CyA (10 mg/kg). The LTX treatment had little effect on the blood concentrations of CyA after i.v. administration, whereas they were decreased significantly after p.o. administration. After p.o. administration, the value of the area under the blood concentration-time curve from 0 to 24 hr and the bioavailability of CyA in the LTX group were decreased to only about one fifth and a quarter of those in the control group, respectively. After treatment with LTX, the expression levels of mdr1a, mdr1b and CYP3A2 mRNAs in the duodenum were markedly increased to about twice the control, but in jejunum, ileum and liver the expression levels were little changed. These findings suggest that the absorption of CyA, which occurs mainly from the upper intestine, is reduced as a result of efflux transport via P-glycoprotein induced by LTX. In conclusion, careful monitoring of CyA levels is required in the event of LTX administration to patients receiving immunotherapy with CyA. PMID- 16272750 TI - Estimation of in vivo percutaneous absorption of emedastine from bile excretion data using a deconvolution method. AB - In vivo percutaneous absorption of emedastine difumarate was investigated in rats and compared with rat skin in vitro. Since emedastine entering the systemic circulation is mostly excreted in bile, we first came up with the method of collecting bile with a minimal skin incision. In vivo skin permeation of the drug was estimated from biliary excretion data by deconvolution analysis. Prior to applying deconvolution analysis, it was confirmed that biliary excretion of emedastine was linear against its dose. When the in vivo permeation profile estimated by deconvolution was compared with the in vitro profile, the lag time for permeation was significantly shorter in vivo than in vitro, whereas the skin permeability coefficient was almost the same. If we presume a two-layer diffusion model, then this finding may primarily be due to the shorter diffusion length of the dermis. PMID- 16272751 TI - Pharmacokinetics and pharmacodynamics of landiolol hydrochloride, an ultra short acting beta1-selective blocker, in a dose escalation regimen in healthy male volunteers. AB - OBJECTIVES: We conducted a randomized, double-blind, placebo-controlled study to evaluate the pharmacokinetics and pharmacodynamics of landiolol hydrochloride in a dose escalation regimen in healthy male volunteers. METHODS: We set two-dose escalation regimen (LM and MH groups) using three different doses [L (low): 0.03 mg/kg/min (1 min) loading-->0.01 mg/kg/min (10 min) continuous, M (medium): 0.06 mg/kg/min (1 min) loading-->0.02 mg/kg/min (10 min) continuous, H (high): 0.125 mg/kg/min (1 min) loading-->0.04 mg/kg/min (10 min) continuous]. Sixteen subjects were allocated randomly to the LM, MH, and placebo groups (n=6, 6, and 4, respectively). RESULTS: In both the LM and MH groups, the blood concentration of landiolol hydrochloride changed within a constant range from 2 minutes after initiation of administration to just before the higher dose escalation. By 2 minutes following the higher dose escalation, the concentration of landiolol hydrochloride reached C(max), and reached almost steady state levels until 6 minutes following administration of the higher dose. The t(1/2) of landiolol hydrochloride was 3.5 minutes. The heart rates and blood pressures of subjects administered landiolol hydrochloride decreased, but there were no adverse events in any subject. CONCLUSIONS: The concentration of landiolol hydrochloride rapidly reached steady state levels, and rapidly dissipated after completion of administration. PMID- 16272752 TI - Novel structure of the CYP2D6 gene that confuses genotyping for the CYP2D6*5 allele. AB - We encountered DNA samples which showed a positive product using a long PCR-based method for the detection of CYP2D6*5, indicating deletion of the entire CYP2D6 gene, but the samples did not show a band related to CYP2D6*5 in either XbaI- or EcoRI-RFLP analysis. To achieve genotyping with accuracy, we performed a further genetic analysis to clarify the discrepancy. An unknown 1.6-kb insert was identified in a region downstream from the CYP2D6 stop codon where a specific primer was designed for long-PCR analysis for CYP2D6*5 genotyping. This finding suggested that the CYP2D6 gene might not be deleted in the samples even if a positive product was detected by the long-PCR method. Furthermore, the allelic frequency of this type was found to be approximately 0.3% (4 heterozygous/771 samples) in a Japanese population. In conclusion, we found a novel structure of the CYP2D6 gene, which might lead to incorrect genotyping for CYP2D6*5. Although the long PCR-based strategy for the detection of CYP2D6*5 has been widely used due to its usefulness and convenience, we recommend caution when adopting this method and propose re-evaluating the method for detecting CYP2D6*5. PMID- 16272753 TI - In vitro inhibitory effect of 1-aminobenzotriazole on drug oxidations in human liver microsomes: a comparison with SKF-525A. AB - 1-Aminobenzotriazole (ABT) is extensively used as a non-specific cytochrome P450 (CYP) inhibitor. In this study, the inhibitory effect of ABT on CYP-dependent drug oxidations was investigated in human liver microsomes (HLM) and compared with that of SKF-525A, another non-specific inhibitor. The following probe activities for human CYP isoforms were determined using pooled HLM: phenacetin O deethylation (CYP1A2); diclofenac 4'-hydroxylation (CYP2C9); S-mephenytoin 4' hydroxylation, (CYP2C19); bufuralol 1'-hydroxylation (CYP2D6); chlorzoxazone 6 hydroxylation (CYP2E1); midazolam 1'-hydroxylation, nifedipine oxidation, and testosterone 6beta-hydroxylation (CYP3A). ABT had the strongest inhibitory effect on the CYP3A-dependent drug oxidations and the weakest effect on the diclofenac 4'-hydroxylation. SKF-525A potently inhibited the bufuralol 1'-hydroxylation, but weakly inhibited chlorzoxazone 6-hydroxylation. The inhibitory effects of ABT and SKF-525A were increased by preincubation in some probe reactions, and this preincubation effect was greater in ABT than in SKF-525A. The remarkable IC50 shift (> 10 times) by preincubation with ABT was observed on the phenacetin O deethylation, chlorzoxazone 6-hydroxylation, and midazolam 1'-hydroxylation. In conclusion, ABT and SKF-525A had a wide range of IC50 values in inhibiting the drug oxidations by HLM with and without preincubation. PMID- 16272754 TI - Stimulatory effects of testosterone and progesterone on the NADH- and NADPH dependent oxidation of 7beta-hydroxy-delta8-tetrahydrocannabinol to 7-oxo-delta8 tetrahydrocannabinol in monkey liver microsomes. AB - Microsomal alcohol oxygenase catalyzes the stereoselective oxidation of 7alpha- and 7beta-hydroxy-delta8-tetrahydrocannabinol (7alpha- and 7beta-hydroxy-delta8 THC) to 7-oxo-delta8-THC in monkey liver, and the activity for 7beta-hydroxy delta8-THC is relatively higher than that for 7alpha-hydroxy-delta8-THC. We previously reported that purified P450JM-E, assumed to be CYP3A8, is a major enzyme responsible for the oxidation of 7-hydroxy-delta8-THC to 7-oxo-delta8-THC in monkey liver and is capable of catalyzing the oxidative reaction by NADH as well as NADPH. In the present study, we demonstrated that some steroids such as testosterone and progesterone stimulated both the NADH- and NADPH-dependent conversions of 7beta-hydroxy-delta8-THC to 7-oxo-delta8-THC in monkey liver microsomes. Kinetic analyses revealed that both the NADH- and NADPH-dependent 7 oxo-delta8-THC formation showed sigmoid kinetics. Testosterone caused a decrease in S50 and an increase in V(max) for the NADH-dependent activity, and resulted in a decrease in S50 without changing the V(max) for the NADPH-dependent activity. On the other hand, NADH-dependent testosterone 6beta-hydroxylation activity showed Michaelis-Menten kinetics and was also inhibited by 7beta-hydroxy-delta8 THC, resulting in a decrease in V(max) with no effect on the K(m). NADPH dependent testosterone 6beta-hydrozylation activity was also inhibited by 7beta hydroxy-delta8-THC, resulting in a decrease in both S50 and V(max). In order to explain the metabolic interaction between 7beta-hydroxy-delta8-THC and testosterone, we propose a kinetic model involving at least three binding sites, for the mechanism of activation by testosterone. PMID- 16272755 TI - Receptor occupancy-based analysis of the contributions of various receptors to antipsychotics-induced weight gain and diabetes mellitus. AB - OBJECTIVE: Among various adverse reactions of atypical antipsychotics, weight gain and impaired glucose tolerance are clinically significant. The aim of this study is to analyze quantitatively the contributions of various receptors to these antipsychotics-induced adverse reactions based on the receptor occupancy theory. METHODS: Two indices of antipsychotics-induced weight gain (the values estimated by a meta-analysis and the observed values in clinical trials) and the morbidity rate of type 2 diabetes mellitus during treatment with antipsychotics were taken from the literature. We calculated the estimated mean receptor occupancies of alpha1 adrenergic, alpha2 adrenergic, dopamine D2, histamine H1, muscarinic acetylcholine (mACh), serotonin 5-HT1A, 5-HT2A and 5-HT2C receptors by antipsychotics by using the pharmacokinetic parameters and receptor dissociation constants, and analyzed the correlation between the occupancies and the extent of adverse reactions as assessed using the aforementioned indices. RESULTS: There were statistically significant correlations between the estimated occupancies of H1 and mACh receptors and antipsychotics-induced weight gain estimated by meta analysis (r(s) = 0.81 and r(s) = 0.83, respectively, p < 0.01). There were also statistically significant correlations between these receptor occupancies and observed weight gain in clinical trials (r(s) = 0.66 in each case, p < 0.01). The morbidity rate of type 2 diabetes mellitus was highly correlated with H1, mACh, and 5-HT2C receptor occupancies (r(s) = 0.90 in each case, p < 0.05). However, H1 receptor occupancy was also highly correlated with mACh receptor occupancy among antipsychotics, so that only one of them may be critically associated with the adverse reactions. Considering that these adverse reactions have not been reported for drugs with mACh receptor antagonistic action, other than antipsychotics, the H1 receptor may contribute predominantly to the antipsychotics-induced weight gain and diabetes mellitus. DISCUSSION/CONCLUSION: Model analysis based on receptor occupancy indicates that H1 receptor blockade is the primary cause of antipsychotics-induced weight gain and diabetes mellitus. PMID- 16272756 TI - Metformin is a superior substrate for renal organic cation transporter OCT2 rather than hepatic OCT1. AB - Although metformin, a cationic agent for type II diabetes, shows its pharmacological effect in the liver, the drug is mainly eliminated into urine. The tissue selectivity based on the function of drug transporters is unclear. In the present study, the transport of metformin was examined using HEK293 cells transiently transfected with five human renal organic ion transporter cDNAs. Human OCT1 and OCT2, but not OAT1, OAT3 or OCT2-A, stimulated the uptake. A kinetic analysis of metformin transport demonstrated that the amount of plasmid cDNA for transfection was also important parameter to the quantitative elucidation of functional characteristics of transporters, and both human and rat OCT2 had about a 10- and 100-fold greater capacity to transport metformin than did OCT1, respectively. In male rats, the mRNA expression level of rOCT2 in the whole kidneys was 8-fold greater than that of rOCT1 in the whole liver. The in vivo distribution of metformin in rats revealed that the expression level of renal OCT2 was a key factor in the control of the concentrative accumulation of metformin in the kidney. These findings suggest that metformin is a superior substrate for renal OCT2 rather than hepatic OCT1, and renal OCT2 plays a dominant role for metformin pharmacokinetics. PMID- 16272757 TI - A novel single nucleotide polymorphism of the human methylenetetrahydrofolate reductase gene in Japanese individuals. AB - The genetic polymorphisms of methylenetetrahydrofolate reductase (MTHFR) have been associated with increased toxicity of methotrexate (MTX), a folic acid antagonist that is widely used to treat cancer and immunosuppressive disorders such as rheumatoid arthritis. In this study, we analyzed all the exons and exon/intron junctions of the MTHFR gene from 200 Japanese individuals. We detected a novel single nucleotide polymorphism (SNP) 148C>T (Arg46Trp) in exon 1. The allele frequency of this polymorphism in the Japanese population appears to be extremely low (0.25%). PMID- 16272758 TI - [Stress and its neural mechanisms]. PMID- 16272759 TI - [The CRH gene as a key player in stress responses]. PMID- 16272760 TI - [Stress-induced gene expression in the hypothalamus]. PMID- 16272761 TI - [A role of the limbico-hypothalamic system in physiological manifestation of stress]. PMID- 16272762 TI - [Stress and animal model of psychiatric disease]. PMID- 16272763 TI - [Perception of stressful stimulus in the prefrontal cortex--the brain mechanism of a failure in adaptation to stress]. PMID- 16272764 TI - [Probes for diagnosis of Alzheimer's disease]. PMID- 16272765 TI - [Recording system for orofacial motor behaviors in the freely behaving mouse]. PMID- 16272766 TI - [Pharmacological and clinical profile of rosuvastatin calcium (Crestor)]. PMID- 16272767 TI - Insulin response to oral glucose loading and coronary artery disease in nondiabetics. AB - Hyperinsulinemia is related to coronary artery disease (CAD), as an indication of decreased insulin sensitivity. Although there are many studies showing the relation between fasting insulin levels and insulin resistance, there are fewer studies on postprandial insulin levels. The aim of the present study was to investigate the relationship between postprandial insulin levels and CAD and its extent in our patients. For this purpose, oral glucose tolerance testing was performed in 222 patients with no known diabetes and who were scheduled to undergo diagnostic coronary angiography. The patients were first separated into two groups, one group (group I) having an insulin response within reference values to oral glucose loading, and the other group (group II) with a higher than normal insulin response. The presence and extent of CAD in the two groups were compared. While 65% of the patients in group 1 had CAD, this rate increased to 79% in group 2 patients (P = 0.02). The mean vessel scores were 0.92 +/- 0.78 in group 1 and 1.67 +/- 0.99 (P < 0.0001) in group 2 patients. The stenosis scores were 2.192 +/- 2.077 in group 1 and 5.588 +/- 3.519 (P < 0.001) in group 2, while the extent scores were 1.230 +/- 1.292 in group 1 and 2.729 +/- 1.847 in group 2 (P < 0.0001). The differences between the two groups were significant. Postprandial insulin values were positively correlated with CAD (P = 0.001, r = 0.214), vessel scores (P < 0.0001, r = 0.326), stenosis scores (P < 0.0001, r = 0.261), and extent scores (P < 0.0001, r = 0.419). Logistic regression analysis revealed hyperinsulinemia increased CAD independent from the other risk factors (OR = 5.742, CI 95%: 1.809-18.227, P = 0.003). PMID- 16272768 TI - Prognostic significance of time-delay to peak creatine kinase after direct percutaneous coronary intervention in acute myocardial infarction patients. AB - The aim of the present study was to investigate the prognostic significance of time-delay to peak creatine kinase (CK) after successful direct percutaneous coronary intervention (PCI) in patients with acute myocardial infarction (AMI). Our 240 consecutive first AMI attack subjects admitted within 5 hours from onset were successfully reperfused by direct PCI therapy. Subjects were divided into two groups according to the upper quartile value of peak-CK time from onset, the early peak-CK group (peak-CK time < or = 16 hours from onset, n = 180) and the late peak-CK group (peak-CK time > 16 hours, n = 60). (I) The early ST-segment resolution rate was lower in the late peak-CK group compared with the early peak CK group (P < 0.05), and there were significantly fewer patients with preinfarction angina pectoris in the late peak-CK group than in the early peak-CK group (P < 0.01). (II) LVEF in the chronic stage was significantly lower in the late peak-CK group than in the early peak-CK group (49 +/- 13% versus 57 +/- 13%, P < 0.001). (III) There were significantly more patients with major complications in the late peak-CK group than in the early peak-CK group (required CABG: 10% versus 3%, P < 0.05; cardiac death: 18% versus 3%, P = 0.0001). (IV) Multivariate analysis identified late peak-CK as an independent predictor of cardiac death (Odds ratio 7.91, 95% C.I. 1.40-44.11, P < 0.05). In patients with AMI, the time delay to peak-CK after successful direct PCI may be closely related to left ventricular systolic dysfunction and poor patient outcome, including mortality. PMID- 16272769 TI - Simultaneous Carotid Endarterectomy and Coronary Revascularization is Safe Using Either On-Pump or Off-Pump Technique. AB - The appropriate surgical strategy for patients with combined carotid and coronary artery disease remains controversial. We retrospectively compared our surgical results for 2 types of approaches in this disorder. The records of 76 patients consecutively operated on for carotid and coronary artery disease between August 1993 and October 2004 were reviewed. There were 18 males (66.6%) and 9 females (33.3%) in group I. Group II consisted of 35 males (71.4%) and 14 females (28.5%). The patients were divided into two groups: patients with combined off pump coronary artery bypass and carotid endarterectomy (group I, n = 27), and those with one-stage on-pump coronary artery bypass and carotid endarterectomy (group II, n = 49). Surgical mortality and morbidity and late outcome were compared among the two groups. The average number of grafts was 1.2 +/- 0.4, with the average operative time of 3.3 +/- 0.3 hours in group I, and 2.3 +/- 0.5 grafts with operative time of 4.6 +/- 0.4 hours in group II (P < 0.001 and P < 0.001, respectively). There was 1 death (3.7%) in group I and 2 deaths (4.8%) in group II (P = 0.937). No patient from either group I or group II had postoperative stroke. Mean hospital stay was 7.4 +/- 1.9 days in group I and 11.3 +/- 1.7 days in group II (P < 0.001). At a mean follow-up of 5.5 +/- 3.3 years in group I, 1 patient had contralateral carotid endarterectomy (3.7%). Group II had a mean follow-up of 5.2 +/- 3.0 years and contralateral carotid endarterectomy was performed in 1 patient (2.0%). There were no late strokes or deaths in either group. Combined coronary artery bypass grafting and carotid endarterectomy using 2 different types of technique is a safe and effective procedure in patients with significant concomitant monolateral carotid and coronary artery disease. PMID- 16272770 TI - C-reactive protein is related to impaired oxygenation in patients with acute aortic dissection. AB - Impaired oxygenation sometimes occurs in patients with acute aortic dissection, however, the mechanism has not been fully investigated. We hypothesized that impaired oxygenation is related to inflammation secondary due to aortic dissection. Patients with acute aortic dissection who had received 14 days of conservative treatment were retrospectively examined. Patients who had undergone surgery or died within 14 days were excluded. Patients who had evidence of having pneumonia or pulmonary congestion during this period were also excluded. Twenty six patients were divided into a preserved oxygenation group (minimum oxygenation index > or = 200, n = 13) and an impaired oxygenation group (minimum oxygenation index < 200, n = 13). Maximum serum C-reactive protein (max CRP) and other factors (age, gender, hypertension, diabetes mellitus, smoking, hyperlipidemia, Stanford type, thrombosed false lumen, pleural effusion, atelectasis, use of intravenous vasodilators) for the two groups were compared. Max CRP was the only predictor for impaired oxygenation as calculated by single/multiple logistic regression analysis. Max CRP was significantly higher in the impaired oxygenation group (20.1 +/- 2.1 mg/dL) than in the preserved oxygenation group (10.5 +/- 1.4 mg/dL, P < 0.05). These results suggest that impaired oxygenation is related to inflammation, which is secondary due to acute aortic dissection. PMID- 16272771 TI - Elevated C-reactive protein levels and increased cardiovascular risk in patients with obstructive sleep apnea syndrome. AB - Obstructive sleep apnea syndrome (OSAS) is associated with cardiovascular morbidity and mortality. Inflammatory processes associated with OSAS may contribute to cardiovascular morbidity in these patients. C-reactive protein (CRP) is an important serum marker of inflammation. We tested the hypothesis that patients with OSAS have increased plasma CRP. After 173 male subjects underwent polysomnography, 94 were considered to have OSAS (apnea-hypopnea index (AHI) > 5), 38 cardiovascular disease (CVD), and 56 without CVD. Seventy-nine subjects were considered not to have OSAS (AHI < 5) and from among these 57 patients without CVD were enrolled as control subjects. Serum CRP levels were significantly elevated in the OSAS + CVD group compared to the two other groups (P < 0.05). When we evaluated the association between the serum CRP level and severity of OSAS, CRP levels were positively correlated with AHI in OSAS patients (r = 0.61, P < 0.001) OSAS, as a marker of inflammation and cardiovascular risk, is associated with elevated levels of CRP. According to these results, the link between cardiovascular morbidity and OSAS may be explained by the coexistence of other cardiovascular risk factors such as circulating CRP levels. PMID- 16272772 TI - Serum lipids in north Indian children treated for kawasaki disease. AB - A prospective cross sectional analysis of serum lipids was carried out in 20 children aged between 1.3 and 16 years with Kawasaki disease (KD). The controls were siblings of other patients. The mean interval between diagnosis of the disease and time of assay was 2.6 years (range 0.41 to 6 years). Standard biochemical methods were employed for determination of various components of the lipid profiles. There were significant differences in high-density lipoprotein cholesterol (HDL-C) (40.37 +/- 12.0 mg/dL versus 53.49 +/- 4.31 mg/dL, P < 0.001) and low-density lipoprotein cholesterol (LDL-C) concentrations (77.76 +/- 26.25 mg/dL versus 57.7 +/- 23.2 mg/dL, P < 0.05) between the cases and controls. Lower HDL-C levels persisted at 1-3 and more than 3 years of disease duration. No significant differences were seen in the values of other parameters in the lipidogram, such as total cholesterol (TC), triglycerides (TG), and very low density lipoprotein cholesterol (VLDL-C). Premature atherosclerosis that occurs in KD may be secondary to these lipid metabolism abnormalities. PMID- 16272773 TI - Factors associated with failure of weaning from long-term mechanical ventilation after cardiac surgery. AB - The purpose of this prospective, quantitative, comparative study, conducted at the 55 bed cardiothoracic intensive care unit of the Heart Institute (InCor), University of Sao Paulo Medical School, was to identify factors involved in the weaning of patients who require long-term (> 10 days) mechanical ventilation after cardiac surgery. The subjects included all patients who underwent open heart surgery with cardiopulmonary bypass during a 10 month period from April 2000 to January 2001 (n = 946). From this group, 52 (5.7%) patients who required a tracheotomy for the management of long-term mechanical ventilation after cardiac surgery with cardiopulmonary bypass were selected. Pre-, intra- and postoperative data from patients who were not successfully weaned after reintubation and who underwent an elective tracheotomy were compared. Parameters of respiratory mechanics such as respiratory complications, oxygenation, and cardiac, renal, and neurological function were evaluated. Weaning success was defined as the ability of a patient to tolerate 48 hours without pressure or flow support from a mechanical ventilator. A patient was considered to have failed weaning if they died or remained under ventilation for more than 8 weeks. Of the 52 patients studied, 25 were successfully weaned, 21 died, and 6 remained ventilated for more than 8 weeks. We found significant statistical differences (P < 0.05) between the groups with respect to success or failure in LVEF (P = 0.0035), the need for vasoactive agents (P = 0.0018), and renal failure (P = 0.002). Parameters of respiratory mechanics and oxygenation (eg, static airway compliance, airway resistance) did not influence the success or failure of weaning. There was a significant difference in relation to the presence of pneumonia (P = 0.0086) between the two groups. Although neurological complications were more frequent in patients in the weaning success group, the failure group had lower GCS scores, which is indicative of worse prognoses. It is concluded that cardiac dysfunction, the need for dialysis, and pneumonia are determinants for weaning failure in patients undergoing long-term mechanical ventilation after cardiac surgery. PMID- 16272774 TI - Ten-year follow-up of familial hypercholesterolemia patients after intensive cholesterol-lowering therapy. AB - To prevent coronary artery disease, it is necessary for patients with familial hyper-cholesterolemia (FH) to maintain a low cholesterol level. Recently a combination therapy of low-density lipoprotein (LDL) apheresis and statins has been used for FH patients, but their long-term prognosis over 10 years is unknown. In this single center prospective report, 18 FH patients with severe coronary stenosis received LDL apheresis every 2 or 4 weeks and statin therapy for 9.8 +/- 3.0 years. Probucol was given to 17 of the 18 patients. We observed their clinical events as well as coronary stenosis findings and ejection fractions for 10.7 +/- 2.6 years. Total and LDL cholesterol levels before therapy were 345 +/- 46 and 277 +/- 48 mg/dL, respectively. Immediately following LDL apheresis, these levels decreased to 104 +/- 7.5 and 66 +/- 16 mg/dL, respectively. There were no cardiac deaths and 4 patients were free from any coronary events. There was one noncardiac death. Nonfatal myocardial in-farction occurred in 2 patients and coronary bypass surgery was required in one patient. Twelve patients received additional coronary angioplasty. There was little change in coronary stenosis and ejection fraction following 10 years of the combination therapy. Univariate Cox regression analysis revealed that the calculated mean LDL cholesterol level was the predictive value of treatment efficacy (mean LDL cholesterol < 140 mg/dL, hazard ratio 0.23, P = 0.028). The combination therapy of LDL-apheresis and antilipid drugs delayed the progression of coronary atherosclerosis and prevented a major cardiac event, although complete inhibition was limited to a small group. Additional coronary angioplasty is likely to be required for a favorable clinical outcome in FH patients. PMID- 16272775 TI - Decreased heart rate recovery in patients with heart failure: effect of fluvastatin therapy. AB - Heart rate recovery is the difference in heart rate at peak exercise and at a specific time interval following the onset of recovery. Attenuated heart rate recovery is an independent predictor of mortality in patients with a history of coronary artery disease. The aim of the present study was to evaluate the effect of a statin on heart rate recovery, particularly in patients with ischemic heart failure and hyperlipidemia. Twenty-nine consecutive hyperlipidemic, stable coronary artery disease patients with heart failure and 19 healthy subjects were enrolled. Heart rate recovery values at the 1st and 3rd minutes and lipid profiles of the patients were evaluated at baseline and following 3 months of treatment with fluvastatin. Compared with healthy subjects, the heart rate recovery values were significantly lower in the heart failure patients in both the 1st and 3rd minutes, respectively (31 +/- 6 versus 19 +/- 7, P < 0.0001; 66 +/- 7 versus 47 +/- 8, P < 0.0001). Heart rate recovery in the 1st and 3rd minutes increased from 19 +/- 7 to 24 +/- 9 and 47 +/- 8 to 57 +/- 11, respectively, following treatment (P < 0.001, P < 0.001). There were no significant correlations among the changes in lipid parameters or HRR in the first and third minutes in the recovery period. The results revealed an improvement in heart rate recovery in heart failure patients by fluvastatin treatment. If this association can be confirmed by other studies, it would be interesting to perform further studies into the mechanism underlying this finding. PMID- 16272776 TI - Impact of the Japanese Diagnosis Procedure Combination-based Payment System on cardiovascular medicine-related costs. AB - In 2003, a lump-sum payment system based on Diagnosis Procedure Combinations (DPC) was introduced to 82 specific function hospitals in Japan. While the US DRG/PPS system is a "per case payment" system, the DPC based payment system adopts a "per day payment." It is generally believed that the Japanese system provides as much of an incentive as the DRG/PPS system to shorten the average length of stay (LOS). We performed an empirical analysis of the effect of LOS shortening on hospital revenue and expenditure under the DPC-based payment system, particularly in cardiovascular diseases. We also point out fundamentally controversial aspects of the current system. A total 109 cases were selected from patients hospitalized at the University of Tokyo Hospital from May to July, 2003 and classified into one of three categories: (1) cardiac catheter interventions, (2) cardiac catheter examinations, and (3) other conservative treatments. We analyzed the changes in profit per day in cases of a reduction in average LOS and an increase in the number of cases. In category (1) profit increased significantly in conjunction with reduced LOS. In category (2) profit increased only minimally. In category (3), profit increased rarely and sometimes decreased. In cases of conservative treatment, profits sometimes decreased because an increase in material costs exceeded the increase in revenue. It therefore became clear that the DPC-based payment system does not decisively provide an economic incentive to reduce LOS in cardiovascular medicine. PMID- 16272777 TI - Increased circulating calcitonin gene-related peptide in congestive heart failure caused by congenital heart disease. AB - Calcitonin gene-related peptide has potent vasodilatory and inotropic actions. The aim of this study was to characterize the changes in this peptide in children with varying degrees of heart failure secondary to congenital heart disease with left to right shunt and to assess its relationship to systolic pulmonary arterial pressure. Plasma calcitonin gene-related peptide levels were measured in 131 children including 13 healthy ones, 43 with various degrees of heart failure secondary to congenital heart disease, and 75 with congenital heart disease without heart failure. In patients with heart failure, calcitonin gene-related peptide concentrations were markedly elevated (15.8 +/- 2.1 pg/mL) as compared with healthy control subjects (7.0 +/- 0.8 pg/mL, P < 0.05) or patients with congenital heart disease but without heart failure (18.6 +/- 1.2 pg/mL, P < 0.01). Compared with the controls, there were highly significant stepwise increases in the calcitonin gene-related peptide levels in the mild (n = 15), moderate (n = 12), and severe (n = 16) heart failure subgroups by 1.5, 1.7 and 3.4 fold, respectively. The plasma calcitonin gene-related peptide levels also correlated directly with the pulmonary arterial systolic pressure (r = 0.515, P < 0.0001). The results of this study indicate that congestive heart failure secondary to congenital heart disease with increased pulmonary flow is associated with elevated levels of calcitonin gene-related peptide that are related to disease severity. Pulmonary overcirculation may play a role in upregulation of calcitonin gene-related peptide in congestive heart failure. PMID- 16272778 TI - Pharmacological preconditioning with bradykinin affords myocardial protection through NO-dependent mechanisms. AB - Bradykinin (BK) is one of the triggers of ischemic preconditioning. Protein kinase C (PKC) and mitochondrial ATP-dependent potassium (K(ATP)) channels are central factors in cardioprotection afforded by BK. However, the role of nitric oxide (NO) in the early phase protection of preconditioning with BK is not well understood. We assessed the signaling pathway of the early phase protection of pharmacological preconditioning afforded by BK. Isolated perfused rat hearts (n = 8/group) were subjected to 30-minute global ischemia and 50-minute reperfusion. Left ventricular systolic pressure (LVSP) was recorded prior to the global ischemia and at the end of reperfusion. Preconditioning with BK was induced by two cycles of 5-minute infusion of BK (0.5 micromol/L) and 5-minute washout prior to the global ischemia. To examine participants in the signaling pathway, 5 hydroxydecanoate (5-HD, 200 micromol/L), chelerythrine (CH, 5 micromol/L), or N(omega)-nitro-L-arginine methyl ester (L-NAME, 50 mmol/L) was added to the perfusate for 5 minutes prior to the infusion of BK. Pharmacological preconditioning by BK improved postischemic recovery of LVSP (+ 45.1% versus control, P < 0.01). Protection by BK was abolished by coadministration of CH, 5 HD, or L-NAME. BK affords myocardial protection in the early phase of pharmacological preconditioning through a pathway that includes endogenous NO, PKC, and mitochondrial K(ATP) channels. PMID- 16272779 TI - The relationship between angiotensin converting enzyme gene polymorphism, coronary artery disease, and stent restenosis: the role of angiotensin converting enzyme inhibitors in stent restenosis in patients with diabetes mellitus. AB - Patients with diabetes mellitus (DM) have advanced atherosclerosis compared with nondiabetics. Restenosis after intracoronary stent implantation occurs frequently in diabetic patients. Angiotensin II is an important growth factor for the development of neointimal hyperplasia after vascular injury. The aim of our study was to evaluate the relationships between angiotensin converting enzyme (ACE) gene insertion/deletion (I/D) polymorphism and coronary artery disease (CAD) and stent restonosis in diabetic patients. One hundred and thirty consecutive patients with CAD and 47 consecutive patients (14 males, mean age, 58.0 +/- 10.0) without CAD were enrolled in the study. All patients had type 2 (noninsulin dependent) DM. The patients with CAD underwent percutaneous transluminal coronary angioplasty (PTCA) and stenting. Ninety-four (59 males, mean age, 60.3 +/- 9.8) underwent control coronary angiography at the end of the follow-up period (mean duration, 9.1 +/- 2.9 months). ACE gene I/D genotyping was identified in all patients. No significant difference was found among patients with and without CAD with respect to ACE gene I/D polymorphism (P = 0.460). In the control coronary angiography, stent restenosis and new lesion development were comparable in each genotyping subgroup. However, a significant relationship was observed between restenosis and the use of ACE inhibitors (ACEI) in patients with D allele (ACEI ratio, 43.5% in the restenosis group and 56.5% in non-restenotic group, P < 0.05). We did not find any relationship between ACE gene I/D polymorphism and CAD and stent restenosis and new lesion development in diabetic patients. On the other hand, ACEI treatment may reduce stent restenosis in type 2 diabetic patients with D allele (DD or ID). PMID- 16272780 TI - A case of atrioventricular nodal reentrant tachycardia with atresia of the coronary sinus ostium. AB - We report a case of atrioventricular nodal reentrant tachycardia coexistent with atresia of the coronary sinus ostium. Radiofrequency current application between the supposed coronary sinus ostium and the tricuspid valve was effective at eliminating the tachycardia. A coronary venogram obtained by left coronary arteriography was useful for guiding the mapping catheter to the successful ablation site. PMID- 16272781 TI - Intracardiac lead fracture in an implantable cardioverter-defibrillator. AB - A patient with an implantable cardioverter-defibrillator (ICD) experienced inappropriate shocks. X-rays revealed that the ventricular lead was fractured at a point, which seemed to be in a mechanical stress-free space in the heart. Multiple leads were entwined making a short flexible section sandwiched between the entwined area and the stiff coil susceptible to the stress from heart beats. The present case shows that indirect concentrated stress caused by restriction of lead flexibility can result in lead fracture. Upon implanting multiple endocardial leads, attention should be given to the relative position of each lead. PMID- 16272782 TI - Inappropriate discharges of intravenous implantable cardioverter defibrillators owing to lead failure. AB - We describe here the case of a 58-year-old female patient who experienced inappropriate shocks from her implantable cardioverter-defibrillator (ICD). Stored electrograms from her ICD showed high frequency noise preceding the shock. Although the pacing threshold was normal and lead fracture was not found in chest X-rays, pacing lead impedance decreased to 480 omega. Moreover, such high frequency noise was observed by electrogram telemetry, but not by routine evaluation every 3 months. ICD lead dysfunction was suspected, so we elected to replace the ICD lead system. At the time of the operation, lead impedance was 410 omega and pacing threshold was the same as it was at the time of the ICD implantation, and no lead insulation disturbances were observed in the generator pocket. However, manipulation of the lead system produced high frequency noise reproducibly. Since some of the ICD lead dysfunction initially was clinically silent at rest, dysfunction was difficult to detect before serious problems occurred. Therefore, more careful evaluation of the ICD lead system is needed during long-term follow-up of ICD implants. PMID- 16272783 TI - Cor triatriatum with repeated episodes of syncope in an eighteen month-old girl: a rare cause of cardiogenic syncope. AB - Syncope can be caused by either cardiogenic or noncardiogenic causes. Structural heart diseases should be considered as a possible cause of recurrent episodes of syncope if no other causes are found. We report an 18 month-old girl who presented with recurrent syncope as well as dyspnea, failure to thrive, and frequent episodes of pulmonary infections, suggesting congestive heart failure. A prominent pulmonic component of the second heart sound and congested lungs noted from chest X-rays indicated pulmonary hypertension. An echocardiogram revealed an abnormal membrane with a small orifice in the left atrium (LA). An intact atrial septum and no evidence of other congenital heart diseases were observed. The result after surgical resection of the membrane was good during follow-up. Cor triatriatum may be one of the structural heart diseases responsible for repeated unexplained syncope. An echocardiogram performed by an experienced cardiologist would facilitate the diagnosis considerably and an excellent prognosis can be achieved via surgical resection of the obstructing membrane. PMID- 16272784 TI - Cough reflex by ventricular premature contractions. AB - A 72-year-old man was referred for further assessment of a chronic cough. He noticed an association between the episodes of coughing and palpitations. Electrocardiography (ECG) revealed normal sinus rhythm and sporadic unifocal ventricular premature contractions (VPCs). Each cough was preceded by a premature beat. Continuous wave Doppler echocardiography revealed a VPC-induced transient increase in the pulmonary artery blood flow. He was successfully treated for VPCs with oral disopyramide, resulting in subsidence of both the coughing and palpitations. We suspect that the VPC-induced hemodynamic changes in the pulmonary circulation might be responsible for coughing in our patient. Premature contractions should be considered as a possible cause of chronic dry cough in the clinical setting. PMID- 16272785 TI - Successful treatment of primary cardiac lymphoma with atrioventricular nodal block. AB - A 69-year-old female suffering from third-degree atrioventricular block with syncope underwent permanent pacemaker implantation. However, she developed shortness of breath 2 months after the implantation. Blood tests revealed elevated levels of LDH, CRP, BNP, and SIL-2R. Transthoracic echocardiography showed thickened left and right atrial walls with mild pericardial effusion. A diagnosis was made based on a CT scan and histology. Although most primary cardiac malignant lymphomas are associated with a poor prognosis, the patient was treated successfully with chemotherapy. PMID- 16272786 TI - A case of takotsubo-shaped hypokinesis of the left ventricle caused by a lightning strike. AB - A 62-year-old woman was struck by lightning while on a mountain and fortunately did not suffer burns or unconsciousness. She stayed at a mountain lodge overnight and was taken to our hospital by helicopter the next day. Upon admission, electrocardiography showed ST segment elevation indicating acute lateral myocardial infarction, and echocardiography showed takotsubo-shaped hypokinesis of the left ventricle indicating an apical aneurysm. Her serum escaped enzyme levels were increased, as is typical in cases of myocardial infarction, however, she did not complain of cardiac symptoms. Coronary arteriography performed 4 days after admission showed a normal coronary artery while left ventriculography showed apical akinesia. An echocardiogram obtained 2 days later showed resolution of the LV wall motion abnormality. This is the first reported case of takotsubo cardiomyopathy caused by lightning. Takotsubo-shaped hypokinesis is not described as a complication of lightning-induced cardiac injury and its pathogenesis remains controversial. PMID- 16272787 TI - Effect of adrenomedullin on adrenergic vasoconstriction in mesenteric resistance arteries of the rat. AB - Adrenomedullin (AM) is a hypotensive peptide that belongs to a family of peptides structurally related to calcitonin gene-related peptide (CGRP). The present study examined the effect of AM on adrenergic nerve-mediated vasoconstriction in rat perfused mesenteric vascular beds without endothelium. Perfusion of AM at 0.1 nM but not 10 nM increased vasoconstrictor responses to periarterial nerve stimulation (PNS) (1-4 Hz), while AM at 10 nM significantly attenuated vasoconstriction induced by bolus injection of norepinephrine (NE). In preparations treated with capsaicin (a CGRP depletor), pressor responses to both PNS and NE injection were markedly attenuated by AM. Perfusion of CGRP(8-37) (a CGRP-receptor antagonist) significantly potentiated the PNS- but not the NE induced vasoconstriction. Combined perfusion of CGRP(8-37) and AM had no effect on the PNS-induced response and antagonized the inhibitory effect of AM on the NE induced response. AM(2-52) (an AM-receptor antagonist) did not influence the effect of AM. These findings suggest that AM facilitates adrenergic vasoconstriction by inhibiting neurotransmission of CGRP-containing nerves, which counteract adrenergic nerve-mediated vasoconstriction. PMID- 16272788 TI - Thrombin-induced force development in vascular endothelial cells: contribution to alteration of permeability mediated by calcium-dependent and -independent pathways. AB - Endothelial cell (EC) barrier dysfunction is associated with many types of vascular diseases. Investigators have hypothesized that altered EC contraction in conjunction with morphological changes may lead to EC dysfunction. However, the nature of EC contraction and its regulatory mechanisms are not fully understood. In this study we measured thrombin-induced force in bovine arterial EC force using EC fibers that were grown in a collagen matrix. Contraction, which occurred in time- and dose-dependent fashion, was elicited by thrombin. The thrombin enhanced EC layer permeability was correlated with EC fiber contraction. These results suggest that EC contractile response is involved in alteration of EC barrier function. During the initial period of thrombin stimulation, cadherin complexes were disrupted and cell-to-cell connections were reduced. This was dependent on the transient increase in intracellular calcium concentration and myosin phosphorylation. Rho kinase activation led to rearrangement of actin stress fibers (ASF). Paracellular holes were created in the EC layer in parallel to EC morphological change. Our findings suggest that EC layer permeability is regulated by two distinguishable steps. In the initial period, the cell-to-cell connection was reduced in a calcium-dependent fashion. Subsequently, Rho kinase and ASF-mediated force development increased EC layer permeability via morphological change of EC. PMID- 16272789 TI - Pathophysiological significance of T-type Ca2+ channels: transcriptional regulation of T-type Ca2+ channel--regulation of CACNA1H by neuron-restrictive silencer factor. AB - Expression of T-type Ca(2+) current in the ventricle varies during development and in cardiac diseases. The alteration in quantity of two isoforms of T-type Ca(2+) channel genes in the heart, CACNA1G and CACNA1H, contributes to the changes of T-type Ca(2+) channel activity. However, the precise mechanisms governing the transcription of T-type Ca(2+) channel genes remain largely unknown. In this review, we briefly describe our recent finding that a transcriptional repressor named neuron-restrictive silencer factor is a potent regulator of T-type Ca(2+) channel gene expression. PMID- 16272790 TI - Pathophysiological significance of T-type Ca2+ channels: expression of T-type Ca2+ channels in fetal and diseased heart. AB - Re-expression of fetal genes has been considered to underlie ionic remodeling in diseased heart. T-type Ca(2+) channels have been reported to be functionally expressed in embryonic hearts. In this review, we summarize developmental changes of T-type Ca(2+) channels in mouse ventricles from 9.5 days postcoitum (dpc) to adulthood, using patch clamp and quantitative PCR. In addition, we introduced T type Ca(2+) channel expression in hypertrophied ventricles caused by myocardial infarction (MI) and aortic banding (AOB). Substantial T-type Ca(2+) channel current was recorded at both 9.5 and 18 dpc. The currents were inhibited by Ni(2+) at low concentrations. The current was not detectable in the adult stage. Ca(v)3.2 (alpha(1H)) mRNA is expressed dominantly at both 9.5 and 18 dpc. Ca(v)3.1 (alpha(1G)) increases from 9.5 to 18 dpc, but remains at low level compared with Ca(v)3.2. In contrast, Ca(v)3.1 is greater than Ca(v)3.2 at the adult stage. In MI, Ca(v)3.1 mRNA correlates negatively with brain natriuretic peptide (BNP) mRNA, whereas Ca(v)3.2 mRNA correlates positively with BNP mRNA. In AOB, these correlations are weak. We also analyzed the neuron-restrictive silencer factor (NRSF) in these hearts because it is the suppressor of transcription of the fetal cardiac gene program. The negative correlation between NRSF and BNP was stronger in MI than in AOB. Our findings show that Ca(v)3.2 underlies the functional T-type Ca(2+) channel in embryonic heart and suggest that NRSF may regulate Ca(v)3.2 expression in diseased hearts. PMID- 16272791 TI - Pathophysiological significance of T-type Ca2+ channels: properties and functional roles of T-type Ca2+ channels in cardiac pacemaking. AB - Calcium channels are essential for excitation-contraction coupling and pacemaker activity in cardiac myocytes. While L-type Ca(2+) channels (LCC) have been extensively studied, functional roles of T-type channels (TCC) in native cardiac myocytes are still debatable. TCC are activated at more negative membrane potentials than LCC and therefore facilitate slow diastolic depolarization in sinoatrial node cells. Recent studies showed that selective inhibition of TCC produced a marked slowing of the pacemaker rhythm, indicating that contribution of TCC to cardiac automaticity was relatively larger than what had been speculated in previous studies. To re-evaluate TCC, we measured current density and kinetics of TCC in sinoatrial node cells of various mammalian species. Current density of TCC was larger in mice and guinea pigs than in rabbit and porcine sinoatrial node cells. Interestingly, few or no obvious TCC were recorded in porcine sinoatrial node cells. Furthermore, it was demonstrated that TCC could be enhanced by several vasoactive substances, thereby increasing spontaneous firing rate of sinoatrial node cells. TCC may, at least in part, account for different heart rates among various mammalian species. In addition, TCC might be involved in physiological and/or pathophysiological modulations of the heart rate. PMID- 16272793 TI - Impaired sympathetic skin response in chronic obstructive pulmonary disease. AB - The sympathetic skin response (SSR) is considered as one of the indexes of autonomic nervous system functions, especially related with the sudomotor function of unmyelinated sympathetic fibers. SSRs are recorded as the potentials with biphasic or multiphasic waveforms by conventional electromyography. SSRs are evaluated by measuring latency (time from the stimulus to the onset), amplitude, and area (the space under the curve of the waveform). Although dysautonomia is a feature of chronic obstructive pulmonary disease (COPD), as demonstrated by acetylcholine sweat-spot test, there are no data concerning SSR in COPD patients. In this study, we electrophysiologically investigated the sudomotor function of the sympathetic nervous system in patients with COPD. SSRs were recorded in 30 patients with COPD and 21 healthy volunteers. Normal responses were obtained from all subjects in the control group. No response was observed in three patients with COPD. The mean latency, amplitude and area values of the potentials recorded of the remaining 27 patients were compared to the control. The mean latency was longer (p<0.01) and the mean amplitude and area values were lower (p=0.012, p=0.021, respectively) in the patients compared to the control. We also demonstrated significant correlations between the latency, amplitude, or area values of the SSR and two parameters of pulmonary function tests forced expiratory volume one second/forced vital capacity (FEV1/FVC) and FEV1/FVC %. In conclusion, SSR is impaired in patients with COPD, which indicates the dysfunction of the sympathetic nervous system. Furthermore, the degree of impairment in SSR may reflect the severity of airway obstruction in patients with COPD. PMID- 16272794 TI - The protective effects of amifostine on adriamycin-induced acute cardiotoxicity in rats. AB - Free oxygen radicals and lipid peroxidation are responsible for adriamycin induced cardiotoxicity. Amifostine is a scavenger of free radicals and may function as a selective cytoprotective agent. The aim of this study was to investigate the effects of amifostine on adriamycin-induced lipid peroxidation and the levels of protective enzymes in the heart. Male Wistar rats were randomly allocated to three groups: pretreated, untreated, and control (n=10 in each group). Rats were pretreated with an intraperitoneal injection of amifostine (200 mg/kg) 30 min before the injection of adriamycin. The pretreated rats were given an intraperitoneal injection of adriamycin (10 mg/kg) and were sacrificed after 72 h. Likewise, rats received intraperitoneal injection of adriamycin (untreated) or saline (control). The hearts were removed for the analyses of malondialdehyde (MDA), reduced glutathione (GSH) and catalase. MDA levels were increased (p<0.005) in the heart tissues of untreated rats compared to control, while GSH and catalase levels were decreased (p<0.05 and p<0.001, respectively) in untreated animals. In amifostine-preatreated group, MDA levels were lower (p<0.01), and GSH and catalase levels were higher (p<0.05 for both) than the untreated group. GSH levels were even higher in the amifostine-pretreated group compared to control (p<0.01), although catalase levels were significantly lower in the pretreated group (p<0.05). These results indicate that amifostine decreases adriamycin-induced lipid peroxidation and increases the levels of the protective enzymes in the heart tissue. Therefore, amifostine may ameliorate the adriamycin-induced acute cardiotoxicity. PMID- 16272795 TI - Forensic assessment of 16 single nucleotide polymorphisms analyzed by hybridization probe assay. AB - Of a number of DNA marker typing techniques for personal identification in the field of forensic medicine, polymorphic short tandem repeat (STR) typing is currently the most frequently used technique. However, the multiplex STR method is time consuming. In contrast, single nucleotide polymorphism (SNP) detection methods are relatively rapid and amenable to high throughput. The discrimination power of each SNP is inferior to that of an STR, but a combination of many SNPs could realize a high discriminating power. In this regard, 16 highly informative SNP markers were selected in the introns of genes whose alleles had a proportion of 0.4-0.6 in the Japanese SNP database. The 16 SNPs were sequentially detected within 40 min using the hybridization probe assay on the LightCycler system. The allele and genotype frequencies of these SNPs were determined in a group comprising 64 unrelated Japanese subjects. Based on the frequency data of this group, the combined matching probability, defined as the estimated probability that two unrelated individuals selected at random would possess identical multilocus genotypes, was calculated with the 16 SNPs in the Japanese population and was found to be 2.025x10(-7). This system is an effective tool in the forensic medicine to obtain information on personal identification. PMID- 16272796 TI - Comparison of brachial-ankle pulse wave velocity in Japanese and Russians. AB - Pulse wave velocity (PWV) is a well-known indicator of arterial stiffness and a marker of the presence of vascular lesions. Cardiovascular mortality in Russia has become the highest in the world. The Japanese are enjoying long lives, and the mortality caused by cardiovascular diseases has thus far remained at lower levels than that in Russia. In this study, we focused on brachial-ankle pulse wave velocity (baPWV) obtained from normal human subjects in Russia as well as in Japan, and compared their respective cardiovascular risks. We evaluated baPWV in 337 Japanese and 138 Russian healthy subjects. The baPWV was recorded using a PWV diagnosis device. BaPWV was measured between 2 locations of the arterial tree. The baPWV in the Russian group was significantly higher than that obtained in the Japanese of two groups categorized by age (40-59 years and 60- years). Further, body mass index (BMI), systolic blood pressure (SBP) and diastolic blood pressure in the Russian group were significantly higher than those obtained in Japanese in three age groups (under 39 years, 40-59 years, and 60- years). Moreover, the baPWV indicated a positive correlation with age, BMI and SBP in both Japanese and Russians, although the increasing trend of the baPWV against age of the Russian group had a larger value than that of the Japanese. Therefore, we suggest that arterial stiffness might be promoted earlier in the Russian group, which might be the main cause of the increased cardiovascular risk in Russia. PMID- 16272797 TI - Concentrations of copper and zinc in liver and serum samples in biliary atresia patients at different stages of traditional surgeries. AB - Many patients with biliary atresia (BA) have impaired metabolism of copper (Cu) and zinc (Zn) because of the obstruction of bile ducts. An excessive Cu accumulation is cytotoxic and results in fibrosis in hepatic tissues. Since Zn works antagonistically to Cu, lower Zn concentrations may deteriorate liver damage. In the 1980's, we performed a series of surgeries on BA patients for the construction and alteration of the bile flow route, which is the major excretion route for Cu. We obtained liver and serum samples at each surgery, and measured Cu and Zn concentrations by inductively coupled plasma atomic emission spectrometry. Hepatic Cu concentration decreased with the improvement of cholestasis after the establishment of bile excretion. Conversely, when cholestasis persisted or recurred, increases in hepatic and serum Cu concentrations were noted. Hepatic Zn concentration was lower than previously reported normal values. High hepatic and serum Cu concentrations due to persistent or recurrent cholestasis and low hepatic Zn concentration may deteriorate hepatic fibrosis and liver cirrhosis. PMID- 16272798 TI - Cystic fibrosis transmembrane conductance regulator gene mutations in infertile males with congenital bilateral absence of the vas deferens. AB - Congenital bilateral absence of the vas deferens (CBAVD) is characterized by azoospermia and male infertility. Mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene are associated with cystic fibrosis (CF), the most common autosomal recessive disorder in Caucasians. Recent publications on CBAVD raised the question whether CFTR gene mutations are responsible for CBAVD occurrence or not. This study was conducted to explore the role of CFTR gene mutations in the occurrence of CBAVD-dependent male infertility. Forty-four chromosomes of 22 CBAVD patients from Austrian ancestry were studied. For detection of the most common mutation DeltaF508, a deletion of phenylalanine at the 508th position of mature CFTR chloride channel protein, the 10th exon of the gene was screened by heteroduplex analysis. In order to identify non-DeltaF508 mutations, we also analyzed the entire coding regions, exon/intron boundaries of 27 exons and the 5'- and 3'-untranslated regions of the gene by denaturing gradient gel electrophoresis (DGGE) after polymerase chain reaction. All exons showing different banding patterns on the DGGE gels were sequenced to define existing DNA sequence variations. Among the analyzed 44 chromosomes of 22 patients, disease producing mutations were found in 31.8% (14/44). The most common mutation was DeltaF508 with a frequency of 43% (6/14), followed by R117H with 29% (4/14). Our results indicate that CFTR gene mutations are common but not the only reason for the occurrence of CBAVD-dependent male infertility. We recommend screening of the CFTR gene in these patients. PMID- 16272799 TI - Helicobacter pylori seroprevalence in patients with mild asthma. AB - Helicobacter pylori (H. pylori) is causally related to chronic active gastritis, peptic ulcer disease, primary low-grade B-cell gastric lymphoma, and is also a risk factor for gastric cancer. In addition, a high seroprevalence of H. pylori has been found in many extragastrointestinal disorders, including active bronchiectasis and chronic obstructive pulmonary disease (COPD). It appears that H. pylori has a close relationship with respiratory diseases, but data in the literature on the relationship between H. pylori infection and asthma are poor. We therefore investigated the relationship between them. In this study we evaluated 46 patients with mild asthma, 48 age- and sex-matched patients with peptic ulcer and 48 healthy control subjects. All enrolled subjects underwent a serologic test for H. pylori IgG and cytotoxin-associated gene-A (CagA) by enzyme linked immunosorbent assay (ELISA). There was no significant difference in both anti-H. pylori IgG seropositivity (p=0.6580) and anti-H. pylori-CagA IgG seropositivity (p=0.7183) between the asthmatic and control subjects. In contrast, both anti-H. pylori IgG seropositivity and anti-H. pylori-CagA IgG seropositivity were significantly higher in peptic ulcer patients than these in asthmatic patients (p<0.01). Despite the sero-epidemiological association of H. pylori infection with many inflammatory conditions, our data show no significant association between mild asthma and H. pylori infection. PMID- 16272801 TI - Does handgrip exercise training increase forearm ischemic vasodilator responses in patients receiving hemodialysis? AB - In patients receiving hemodialysis, exercise capacity is extremely limited. Although vasodilation is one of the key phenomena for blood perfusion into working skeletal muscles during exercise, it is not clear whether the vasodilator capacity is increased after physical training in this population. We attempted to clarify whether handgrip exercise training increases forearm vasodilator responses to arterial occlusion, and to determine the relationship between muscle contraction function and the vasodilator responses in patients receiving hemodialysis. Eight patients and 7 age-matched healthy controls were tested. The patients participated in handgrip training four times a week for 6 weeks. Before and after the training the maximum muscle strength and endurance were measured with a handgrip dynamometer, and the forearm vasodilator responses to 3-minute arterial occlusion were measured by the near infrared spectroscopy technique. Maximum strength and endurance were significantly lower in the patients than in the controls. Maximum strength (from 183+/-84 to 228+/-92 Newtons, p<0.05) and endurance (from 19+/-6 to 31+/-8 sec, p<0.05) were both increased after the training in the patients. Vasodilator responses estimated by the ratio of the maximum value of oxyhemoglobin after relief of arterial occlusion to its minimum value before the relief were significantly smaller in the patients compared with those in the controls (132+/-20 vs 161+/-27%, p<0.05). In contrast to the findings in muscle function, the decreased vasodilator responses were not improved after the training (141+/-17%). Additionally, no improvement in the vasodilator responses was observed in the parameters estimated by oxygen saturation. These data suggest that exercise capacity increased by physical training is produced by the functional improvement of skeletal muscles per se, but not by alterations in blood perfusion for oxygenation of the muscles in patients receiving hemodialysis. PMID- 16272800 TI - Inflammatory mediators down-regulate 11beta-hydroxysteroid dehydrogenase type 2 in a human lung epithelial cell line BEAS-2B and the rat lung. AB - In the lung, anti-inflammatory actions of glucocorticoids would be determined by 11beta-hydroxysteroid dehydrogenase type 2 (11beta-HSD2), the microsomal enzyme responsible for the breakdown of bio-active glucocorticoids. However, regulation of 11beta-HSD2 under inflammatory conditions such as acute lung injury is not well understood. In the present study, we examined whether inflammatory substances would influence the activity and mRNA expression of 11beta-HSD2 in the lung. In a human bronchial epithelial cell line BEAS-2B, endotoxin inhibited 11beta-HSD2 enzyme activity in a dose-dependent manner over 48 h with a significant decrease in the mRNA expression. Likewise, tumor necrosis factor (TNF)-alpha inhibited both activity and mRNA expression of 11beta-HSD2. The TNF alpha-dependent decrease in the enzyme activity was completely blocked by anti TNF-alpha antibody, while antibody alone showed no significant influence on the enzyme activity. An nitric oxide donor (NO) sodium nitropusside or a cGMP analog 8-br-cGMP caused moderate but significant decreases in both activity and mRNA expression of 11beta-HSD2. Importantly, treatment of rats with endotoxin significantly decreased both activity and mRNA expression of 11beta-HSD2 in the lung tissue. We conclude that lung inflammation reduces local glucocorticoid breakdown and augments glucocorticoid action in the lung by down-regulating 11beta-HSD2 via multiple mechanisms. PMID- 16272802 TI - Environmental exposure and p53 mutations in esophageal cancer patients in areas of low and high incidence of esophageal cancer in China. AB - Esophageal cancer is the 6th most common cancer in the world, and genetic factors (p53 mutations) in addition to the environmental factors (food, nutrition, smoking, drinking, etc.) are involved in its development. In this study, the association between the both factors, environmental risk factors for esophageal cancer and p53 mutations, in tumor tissues was investigated in 77 patients living in a high-incidence area and 50 patients living in a low-incidence area in Hebei Province, China. Among these patients, p53 mutations were observed in about 50%, without regional differences in the respective frequencies. G:C>A:T (G to A or C to T) transition mutations were the major type of mutations observed in patients in the high-incidence area (19 patients, 50%), whereas G:C>A:T transitions and insertions were observed with equal frequency (8 patients, 33.3%) in the low incidence area. As for the association with environmental factors, p53 mutations occurred with higher frequency in patients with a daily intake of spicy foods and in those who used unboiled well water in the low-incidence area. Logistic regression analysis showed no association between food intakes and p53 mutations in high- and low-incidence areas. Thus, higher frequency of spicy food intake and use of unboiled well water may be risk factors of esophageal cancer via p53 mutations in China. PMID- 16272803 TI - Gender and family composition related to discharge destination and length of hospital stay after acute stroke. AB - Informal care by family members still plays an important role in home care after acute stroke. This study determined the clinical and demographic factors, such as family structure, that predict discharge to home and length of hospital stay (LOS) after acute stroke hospitalization. We reviewed the sex, age, family structure before stroke, type of stroke, size of the lesion, activities of daily living (ADL) function at discharge, discharge destination, and LOS of stroke patients (114 cerebral infarctions and 44 intracerebral hemorrhages) admitted to a neurosurgical hospital. Patients with cerebral infarction were older than those with intracerebral hemorrhage (median 75 vs 66 years). Ninety-eight were discharged to home (62%). In the logistic regression analysis, low ADL function, medium or large infarction, and intracerebral hemorrhage (vs lacunar infarction) were significantly associated with discharge to a destination other than home. Of the patients discharged home, low ADL function was strongly associated with LOS in the multiple regression analysis. In addition, living with a spouse only had the opposite effect on LOS in men and women (p=0.050 and 0.071, respectively). LOS tended to be shorter for men with a wife, but longer for women with a husband. The structure and gender roles in a stroke patient's household may need further attention for the efficient use of hospital resources. PMID- 16272804 TI - Long-acting gonadotropin-releasing hormone analogue treatment for central precocious puberty in maternal uniparental disomy chromosome 14. AB - Uniparental disomy (UPD) is the inheritance of a chromosome pair from one parent and is increasingly recognized as a cause of abnormal phenotypes either due to imprinted genes or, in the case of isodisomy, to homozygosity of recessive alleles. Maternal uniparental disomy for chromosome 14 (matUPD[14]) may cause a characteristic phenotype including precocious puberty. Central precocious puberty (CPP) was diagnosed in a 6-year-old girl with some dysmorphic features, truncal obesity, small hands, and small feet. Cytogenetic analysis of her peripheral blood demonstrated chromosomal rearrangement: Robertsonian translocation 45, XX, der(13;14)(q10;q10). MatUPD(14) was demonstrated in the patient by haplotype analysis of chromosome 14, showing that the CPP is one of the features caused by matUPD(14). The CPP was successfully treated with higher dosage of long-acting gonadotropin releasing hormone (GnRH) analogue, Leuprolide, 90 microg/kg/month. This is the first report that describes GnRH analogue treatment for CPP associated with matUPD(14), suggesting that the GnRH analogue treatment is appropriate even for such a specific type of CPP. PMID- 16272805 TI - [Need to change the direction of cholesterol-related medication--a problem of great urgency]. AB - The cholesterol hypothesis implies that reducing the intake of saturated fatty acids and cholesterol and increasing that of polyunsaturated fatty acid are effective in lowering serum total cholesterol (TC), and thereby reducing the incidence of coronary heart disease (CHD). However, these dietary recommendations are essentially ineffective in reducing TC in the long run, but rather increase mortality rates from CHD and all causes. The reported "apparent relative risk of high TC in CHD mortality" (the ratio of mortality at the highest/lowest TC levels) varied several-fold among populations studied. The incidence of familial hypercholesterolemia (FH) in a population was proposed to be a critical factor in the observed variability, which could be accounted for by assuming that 1) the high CHD mortality rate in high-TC groups is mainly a reflection of the incidence and severity of FH, and 2) high TC is not a causative factor of CHD in non-FH cases. This interpretation is supported by recent observations that high TC is not positively associated with high CHD mortality rates among general populations more than 40-50 years of age. More importantly, higher TC values are associated with lower cancer and all-cause mortality rates among these populations, in which relative proportions of FH are likely to be low (circa 0.2%). Although the effectiveness of statins in preventing CHD has been accepted in Western countries, little benefit seems to result from efforts to limit dietary cholesterol intake or to TC values to less than approximately 260 mg/dl among the general population and the elderly. Instead, an unbalanced intake of omega6 over omega3 polyunsaturated fats favors the production of eicosanoids, the actions of which lead to the production of inflammatory and thrombotic lipid mediators and altered cellular signaling and gene expression, which are major risk factors for CHD, cancers, and shorter longevity. Based on the data reviewed here, it is urgent to change the direction of current cholesterol-related medication for the prevention of CHD, cancer, and all-cause mortality. PMID- 16272806 TI - [In-silico prediction of pharmacokinetic properties]. AB - In silico methods for predicting pharmacokinetic properties range from data-based approaches such as quantitative structure-activity relationships (QSARs), similarity searches, and 3-dimensional QSAR, to structure-based methods such as ligand-protein docking and pharmacophore modelling. Data-based modelling approaches are effective for many drug absorption, distribution, metabolism, and excretion (ADME) processes such as passive membrane permeation, where their molecular mechanism is barely delineated. Therefore QSAR approaches have been applied to simulate the relationships between ADME parameters and molecular structure and properties. In the present investigation, we describe the application of the genetic algorithm-combined partial least-squares (GA-PLS) method to QSAR modelling of various ADME properties. By selecting an appropriate set of molecular descriptors automatically using the genetic algorithm, many ADME properties could be well explained by simple molecular descriptors derived from the 2-dimensional chemical structure. PMID- 16272807 TI - [Synthesis of 2-, 4- and 5-(2-alkylcarbamoyl-1-methylvinyl)-7 alkyloxybenzo[b]furans and their leukotriene B4 receptor antagonistic activity]. AB - Variable 7-carboxylpropoxy or (1-phenyl)ethoxybenzo[b]furan derivatives with (E)- and (Z)-2-alkylcarbamoyl-1-methylvinyl groups at the 2-, 4-, and 5-positions were prepared to find novel and selective leukotriene B(4) (LTB(4)) receptor antagonists. (E)-2-(2-Diethylcarbamoyl-1-methylvinyl)-7-(1 phenylethoxy)benzo[b]furan (4v) showed selective inhibition of the human BLT(2) receptor (hBLT(2)). On the other hand, (E)-2-acetyl-4-(2-diethylcarbamoyl-1 methylvinyl)-7-(1-phenylethoxy)benzo[b]furan (7c) inhibited both human BLT(1) receptor (hBLT(1)) and hBLT(2). The (E)-2-(2-diethylcarbamoyl-1-methylvinyl) group lay on approximately the same plane as the benzo[b]furan ring, whereas the (E)-4-(2-diethylcarbamoyl-1-methylvinyl) group had a torsion angle (45.7 degrees ) from the benzo[b]furan ring plane. However, the (Z)-(2-alkylcarbamoyl-1 methylvinyl)benzo[b]furans were inactive. The inhibitory activity depended on the conformation of the 2-alkylcarbamoyl-1-methylvinyl groups. PMID- 16272808 TI - [Comprehensive determination of furanocoumarin derivatives in citrus juice by high performance liquid chromatography]. AB - We studied a reverse phase HPLC method employing a simple acetonitrile: 0.1% phosphoric acid aqueous solution gradient as the mobile phase for the determination of furanocoumarin (FC) derivatives, such as bergamottin (BG) and 6',7'-dihydroxybergamottin (DHB), using UV detection. Anthracene was added to samples as an internal standard. A Capcell Pak SG-Phenyl column (4.6 mm [inner diameter] x 25 cm; particle size 5 mm; Shiseido) was used, and the flow rate was set at a constant 1 ml/min. A photodiode array detector was used because it reveals the characteristic UV-absorption spectrum of FCs, commonly with 311 nm as the maximum wavelength. Furanocoumarin derivatives in pomelo juice (PJ) were detected by the method and compared with those in grapefruit juice (GJ) and orange juice (OJ). GJ contained 3 kinds of FCs, BG, DHB and bergaptol (BT). OJ had no FCs. On the other hand, PJ contained 8 kinds of FCs including BT, BG and DHB. This FC detection system may be effective for identifying foods and beverages that interact adversely with drugs. PMID- 16272810 TI - [Effects of sweetie juice on nifedipine pharmacokinetics in rats]. AB - It has been reported that grapefruit juice (GJ) causes pharmacokinetic interactions with many drugs after co-ingestion, but the effects of the juice of sweetie fruit, a cross between a pomelo and a grapefruit, on the pharmacokinetics of medicines have not been clear. The present study investigated the drug interaction capability of sweetie juice (SJ). The effect of SJ on nifedipine (NFP) pharmacokinetics in rats was studied. Two milliliters of SJ, GJ, or saline was administered to the rat duodenum. After 30 min, NFP was administered intraduodenally at a dose of 3 mg/kg body weight. The NFP concentrations in the plasma samples were determined by high performance liquid chromatography (HPLC). Although GJ increased the area under the plasma concentration-time curve (AUC) of NFP (1.6-fold), SJ had no significant effect on the NFP pharmacokinetics in rats. Furthermore, concentrations of furanocoumarin derivatives in SJ were measured by HPLC equipped with a photodiode array detector, and compared with those in GJ. SJ contained lower concentrations of bergamottin (0.53 microg/ml), 6', 7' dihydroxybergamottin (0.19 microg/ml), and bergaptol (0.2 microg/ml) than the GJ used in this study (6.3 microg/ml, 3.6 microg/ml, and 9.4 microg/ml, respectively). In conclusion, the results suggest that SJ had no effect on the NFP pharmacokinetics in rats due to low furanocoumarin concentrations in SJ. PMID- 16272809 TI - [Examination of rheumatoid factor and other serum markers in rheumatoid arthritis]. AB - Rheumatoid factor (IgM-RF) has been widely used to diagnose rheumatoid arthritis (RA) in clinical practice. We investigated the RA diagnostic performances of anti cyclic citrullinated peptide antibody (anti-CCP), matrix metalloproteinase-3 (MMP 3), anti-agalactosyl IgG antibody (CA*RF), and anti-calpastatin antibody (ACA) in comparison with IgM-RF. Among 68 RA patients, IgM-RF was positive in 31 (45.6%) and negative in 37 (54.4%). In the IgM-RF-positive group, positivity in anti-CCP, CA*RF, and ACA was 97%, 100%, and 97%, respectively, although that in MMP-3 (74%) was inferior to the others. On the other hand, in the IgM-RF-negative group, positivity in anti-CCP, MMP-3, and ACA was 73%, 81%, and 86%, respectively, although that in CA*RF was only 59%. We conclude that the combination of IgM-RF and anti-CCP/ACA will provide an accurate diagnosis of RA in clinical practice. PMID- 16272811 TI - Introduction of grapes and alfalfa into China: a reflection on the role of Zhang Qian. AB - There is a prevailing legend that grapes and alfalfa were introduced to China from West Asia by Zhang Qian in 126 BC. This paper intends to show the legend to be a misconception. The three crucial documents were thoroughly examined, and the author believes he can demonstrate that it was a later traveler who actually brought these two plants to China following the trade route pioneered by Zhang Qian. PMID- 16272812 TI - Effect of hot-smoked rainbow trout (Oncorhynchus mykiss) diets on the protein levels of liver tissue in rats. AB - AIMS: Free radicals may not only affect the denatured cell structure but also reduce protein levels. These mechanisms are critical and need to be clarified as a high priority. Because of the limited information on this subject, in this respect, the effects on protein levels of liver tissue in rats fed hot-smoked rainbow trout (Oncorhynchus mykiss) were investigated. METHODS: Four diets containing fresh and hot-smoked rainbow trout flesh and vitamins were prepared and commercial pellet food was purchased. Four groups of female Wistar rats were fed the diets for 28 days. The rats were killed by decapitation, and the hepatic tissue was removed. Total protein and detection of protein bands using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) were evaluated. RESULTS: The liver protein level in rats fed a hot-smoked rainbow trout flesh + vitamin diet was increased significantly compared with the other diet groups (p < 0.001, p < 0.009 and p < 0.011). 10 protein bands were visualized on rat gels. The molecular weights of protein bands detected were 200, 130, 70, 50, 42, 38, 26, 25, 24 and 14 kDa, respectively. The 25-kDa band of group V had a dense appearance. Also, the 14-kDa band appeared as very dense in groups V, B and K, but this band did not have a dense appearance in group D. CONCLUSION: Nourishment with smoked fish diets containing antioxidant substances reduced the harmful effects of free radicals and positive effects were observed on protein metabolism. PMID- 16272813 TI - Tissue electric properties in head and neck cancer patients. AB - BACKGROUND: The phase angle of the impedance vector was lower because of a decreased Xc component in hemodialysis patients with poorer prognosis, patients with hemodynamic instability, and in critically ill patients. The phase angle is easy to obtain. The aim of our study was to investigate in a case-control study the utility of phase angle and other impedance parameters in a population of male patients with head and neck cancer. MATERIAL AND METHODS: A case-control study was designed. A population of 67 ambulatory post-surgical male patients was enrolled with the following inclusion criteria: oral and/or laryngeal cancer confirmed by biopsy, without a recent loss weight (<5% during previous 3 months) and signed informed consent. As reference group, we selected 70 male subjects from the same geographic region and matched by age, which were selected from a database of healthy people of our hospital. Basal blood sampling was performed for determinations of blood chemistry. Weight, height, body mass index and tetrapolar body electrical bioimpedance were performed in both groups. RESULTS: A total of 67 head and neck cancer male patients were enrolled, mean age was 58.49 +/- 14.54 years, weight 73.32 +/- 11.4 kg and BMI 28.53 +/- 3.5. A total of 70 controls subjects were studied, mean age was 62.33 +/- 12.4 years. Weight 64.31 +/- 8.38 kg and BMI 24.33 +/- 3.2 were significantly lower than in cancer patients (p < 0.05). Renal function and blood sodium levels were similar in both groups. Anthropometric evaluation in cancer patients showed a mean fat mass lower than control patients (13.9 +/- 6.1 vs. 12.1 +/- 6.1 kg; p < 0.05). In control patients, fat-free mass was higher than in cancer patients (58.7 +/- 8.2 vs. 51.23 +/- 8.4 kg; p < 0.05). In cancer patients, reactance (62.3 +/- 17.2 vs. 56.6 +/- 15.1 ohm; p < 0.05) and phase angle (8.02 +/- 1.3 vs. 6.9 +/- 1.5 degrees ; p < 0.05) were lower than in control patients. CONCLUSION: Impedance in male head and neck cancer patients were characterized by a reduced reactance and phase angle. These early altered tissue electric properties appeared with a normal weight and body mass index. PMID- 16272814 TI - An appropriate indication for the initiation of beta-blocker therapy in dilated cardiomyopathy. AB - BACKGROUND: Although long-term treatment with beta-blockers has been shown to improve morbidity and mortality in dilated cardiomyopathy (DCM), patient responses are heterogeneous. METHODS: To establish the appropriate indication for the initiation of beta-blocker therapy, we retrospectively analyzed 38 DCM patients treated with beta-blockers (metoprolol or carvedilol) and examined differences in baseline profiles between patients who could continue the therapy (responders) and those who could not (non-responders). RESULTS: In 13 non responders, the duration from onset of symptoms to beta-blocker initiation was longer (p < 0.05), systolic blood pressure was lower (p < 0.001), serum sodium concentration was lower (p < 0.05), left ventricular posterior wall thickness was thinner (p < 0.05), left ventricular end-diastolic pressure was higher (p < 0.05) and left ventricular wall stress was lower (p < 0.05) than in 25 responders. In 19 patients receiving carvedilol, 5 non-responders showed higher levels of human atrial natriuretic peptide (p < 0.05) and brain natriuretic peptide (p < 0.01) than 13 responders. Discriminant analysis with a linear discriminant function showed the following equation predicted response to beta-blocker therapy: h = 0.004 x systolic blood pressure - 0.002 x brain natriuretic peptide + 0.667 (R2 = 0.67, p < 0.001). The probability of predicting the response was 94.1% with h > or = 0.5. CONCLUSION: We conclude that h > or = 0.5 is the appropriate indication for the initiation of beta-blocker therapy in DCM. PMID- 16272815 TI - Longitudinal variance of fat mass deposition during pregnancy evaluated by ultrasonography: the ratio of visceral fat to subcutaneous fat in the abdomen. AB - BACKGROUND: Defining regional fat distribution is of importance because of regional variation in adipocyte metabolism. In order to investigate whether maternal body fat distribution changes during pregnancy we measured the abdominal maternal fat thickness. METHODS: A longitudinal study of the thickness of the preperitoneal fat layer (P) and subcutaneous fat layer (S) in the abdomen was conducted; ultrasonography was used to obtain the measurements and the P/S ratio was calculated during each trimester as well as postpartum. RESULTS: When compared with the first and second trimesters, a significant increase in both the P and P/S ratios was observed in the third trimester; thus, regional fat distribution changes during pregnancy. CONCLUSION: Intra-abdominal visceral fat accumulation increases during pregnancy. PMID- 16272816 TI - Value of endocervical ureaplasma species colonization as a marker of preterm delivery. AB - AIMS: To study the predictive value for preterm delivery of colonization of the cervix and vagina by ureaplasmas and other potentially pathogenic microorganisms. METHODS: Prospective analysis of a study group of 200 pregnant women with preterm labor and intact membranes, and a control group of 50 pregnant women. The subjects in both groups were between 24 and 34 weeks of gestation. Ureaplasma spp. and Mycoplasma hominis endocervical cultures were performed for both groups. RESULTS: In the study group, 70 of the women delivered preterm, and all women in the control group carried their pregnancies to full term. Cervical cultures for Ureaplasma spp. were positive in 119 women, and were isolated more frequently in the study group (51.5%) than in the control group (32%), and more frequently in subjects in the study group who delivered preterm (65%) than in those who had a full-term delivery (45%), differences that were statistically significant. CONCLUSION: Detection of Ureaplasma spp. in the endocervix has a statistically significant relationship to preterm labor (p = 0.03) and preterm delivery (p = 0.02) in pregnant women with preterm labor and intact membranes. PMID- 16272817 TI - Plasma cell membrane glycoprotein-1 K121Q polymorphism in preeclampsia. AB - BACKGROUND: Preeclampsia is a common hereditary disease with unclear aetiology and various genetic and environmental components. We wanted to determine whether genetic variability in the gene encoding plasma cell membrane glycoprotein-1 (PC 1) contributes to individual susceptibility to the development of preeclampsia. METHODS: The case-control study involved 133 women with preeclampsia and 115 healthy controls. They were genotyped for the K121Q polymorphism in the PC-1 gene. chi(2) analysis was used to assess genotype and allele frequency differences between preeclamptic and control women. RESULTS: The frequency of the PC-1 gene 121K allele was found to be equal in the two groups, being 90.2% among women with preeclampsia and 90.4% among controls (p = 0.937; OR = 1.024; 95% CI = 0.564-1.861). Also the genotype distribution of the PC-1 K121Q polymorphism was similar (p = 0.516) in the preeclamptic and control groups. CONCLUSIONS: The K121Q polymorphism of the PC-1 gene is unlikely to be a major genetic factor predisposing to preeclampsia in Finnish women. PMID- 16272818 TI - Lipid peroxidation and Ca-ATPase activity of basal plasma membranes of syncytiotrophoblast from normotensive pregnant women. AB - BACKGROUND: The Ca-ATPase activity of the plasma membranes of several tissues of preeclamptic pregnant women is significantly reduced when compared with the values of normotensive pregnant women. This has been explained considering the raise in the level of lipid peroxidation of the plasma membranes with preeclampsia. In this work we studied the effect of lipid peroxidation of syncytiotrophoblast basal (fetal facing) plasma membranes from normotensive pregnant women, on their level of Ca-ATPase activity. METHODS: The syncytiotrophoblast basal (fetal facing) plasma membranes from normotensive pregnant women were isolated and irradiated with ultraviolet (UV) light (254 nm). The membranes were then assayed for Ca-ATPase activity and lipid peroxidation by TBARS. RESULTS: The UV irradiation raises the level of lipid peroxidation of the membranes, producing a concomitant inhibition of their Ca-ATPase activity. Presence of the antioxidant butylated hydroxytoluene during the UV irradiation of the membranes prevents increase in their level of lipid peroxidation and hence the inhibition of their Ca-ATPase activity. CONCLUSION: These results give a strong support to the hypothesis that the lowered Ca-ATPase activity already described for plasma membranes of several tissues of preeclamptic women is the consequence of the increased level of lipid peroxidation shown by these membranes. PMID- 16272819 TI - A patient with hypopituitarism and isochromosome 18q mosaicism. AB - AIMS: Patients with isochromosome 18 [i(18q)] have features of both trisomy 18 and deletion of 18p [del(18p)] syndromes. Although, hypopituitarism has been reported in patients with del(18p) syndrome, it has not been described in patients with i(18q) syndrome previously. We describe a case with i(18q)/del(18p) mosaicism associated with a novel finding of hypopituitarism. METHODS: Clinical characteristics of the patient have been discussed in the light of the literature. RESULTS: The patient had dysmorphic findings that are predominantly seen in del(18p) syndrome such as low nasal bridge, wide mouth, large ears, high forehead, hypopigmentation, upturned nostrils and hypopituitarism (TSH, ACTH, and GH deficiencies, and pituitary hypoplasia on magnetic resonance imaging). In addition, she also had upturning of upper lip and seizures, which are features of trisomy 18 syndrome. CONCLUSIONS: In agreement with the previous clinical reports, this case further supports the presence of a factor, which is involved in pituitary development and/or function, on the short arm of chromosome 18. PMID- 16272820 TI - Differential effects of short-acting beta2-agonists on human granulocyte functions. AB - BACKGROUND: beta2-Adrenergic agonists play a pivotal role in the management of bronchial asthma. Although the major effect of short-acting beta2-agonists on the airway is relaxation of smooth muscles, they may also have several effects on surrounding immunomodulatory cells. METHODS: We examined whether widely used short-acting beta2-agonists differ in their ability to modulate granulocyte functions, such as superoxide anion (O2-) production and degranulation. RESULTS: Procaterol (PC), a full agonist, significantly inhibited both O2- production by granulocytes (neutrophils and eosinophils) and their degranulation at the clinically relevant concentrations, whereas salbutamol and tulobuterol (partial agonists) showed smaller effects. PC inhibited N-formyl methionyl-leucyl phenylalanine-induced O2- production and peroxidase release, but failed to inhibit responses induced by phorbol 12-myristate 13-acetate and/or opsonized zymosan. Exposure to 5 x 10(-8)M PC for 120 min resulted in approximately 50% inhibition of O2- production and degranulation of neutrophils. The effects of beta2-agonists were more obvious in neutrophils than in eosinophils. A selective beta2-receptor antagonist, ICI-118551, reversed the inhibitory effect of beta2 agonists (PC, salbutamol, tulobuterol B) on N-formyl methionyl-leucyl phenylalanine-induced O2- production. CONCLUSIONS: These results suggest that beta2-agonists had an inhibitory effect on granulocyte functions, mainly mediated viareceptors and their efficacy. Our observations support that beta2-agonists with a rapid onset of action and high intrinsic efficacy (short-acting and full agonists) may be optimal for the rescue therapy against acute asthma attack and sedation of its airway inflammation in an early phase. PMID- 16272822 TI - Excretory-secretory products produced by paragonimus westermani differentially regulate the nitric oxide production and viability of microglial cells. AB - BACKGROUND: Tissue-invading helminth parasites secrete a large amount of cysteine proteases that may play critical roles in tissue invasion and immune modulation. However, roles of excretory-secretory products (ESP) secreted by Paragonimus westermani in the activation and death of microglial cells in brain are poorly understood. OBJECTIVES: In the present study, we investigated whether ESP could regulate microglial nitric oxide (NO) production and viability. METHODS: The NO production and cell viability were assessed by respectively measuring the formation of nitrite and the release of lactate dehyrogenase. RESULTS: At a low (0.2 microg/ml) concentration, ESP significantly stimulated NO production with no apparent cell injury or death in cultured microglial cells. However, at high (> or =2 microg/ml) concentrations, ESP induced severe cell death. Inhibition of inducible NO synthase significantly reduced the NO productivity, but not cytotoxicity, of ESP. Similarly, inhibitors of the extracellular signal-regulated kinase, p38 and nuclear factor kappa B also blocked only the NO productivity of ESP. Interestingly, heat inactivation did not hamper the ability of ESP to stimulate microglial NO production. Similarly, pretreatment with thiol crosslinking reagents dramatically reduced both proteolytic activity and cytotoxicity of ESP, but did not alter NO production in microglial cells. Interestingly, although cysteine protease competitive inhibitors and thiol alkylating reagents markedly reduced the proteolytic activity of ESP, they did not influence the NO productivity and cytotoxicity of ESP. CONCLUSION: The present results indicate that the NO production and cytotoxicity by ESP may be differentially regulated via unknown mechanisms, not related with cysteine protease activity. PMID- 16272821 TI - Allergy immunotherapy as an early intervention in patients with child-onset atopic asthma. AB - BACKGROUND: In patients with bronchial asthma an effective treatment is required at early stages of the disease to prevent irreversible structural changes of the airways. The objective of this study was to evaluate the beneficial effects of our routine immunotherapy as an early intervention on FEV1 in patients with child onset atopic asthma. METHODS: Beneficial effects of successful immunotherapy on FEV1 were analyzed retrospectively in 43 unselected patients who received our routine standard subcutaneous immunotherapy with periodic FEV1 measurements and became asymptomatic. RESULTS: Although there was no significant correlation between the duration of asthma symptoms prior to immunotherapy and the changes in FEV1 before and after immunotherapy in 43 unselected patients, there was a significant inverse correlation between these two parameters in 23 patients whose asthma duration was less than 20 years. As the FEV1 increased after immunotherapy in all 14 patients whose asthma duration was less than 5 years, the 43 patients were divided into a group 1 including these 14 patients and a group 2 including 29 patients whose asthma duration was more than 5 years. The FEV1 decreased in 7 of the 29 asymptomatic patients in group 2. There was no difference in the initial FEV1 between the two groups, but the final FEV1 and the mean of the average increase in FEV1 per year by immunotherapy were significantly higher in group 1 than in group 2. CONCLUSIONS: Immunotherapy should be started as early as possible at the youngest age in order to increase a beneficial effect of successful immunotherapy on FEV1 improvement. PMID- 16272823 TI - Lack of association between the IL13 variant Arg110Gln and susceptibility to cedar pollinosis in a Japanese population. AB - BACKGROUND: Interleukin (IL)-13 has come to be appreciated as a molecule critically involved in allergic inflammatory responses. Recent studies revealed that a common variant in the coding region of the IL13 gene, Arg110Gln, has been implicated in the development of asthma and atopy. METHODS: To assess whether the IL13 variant Arg110Gln is associated with cedar pollinosis, one of the most common atopic diseases in the Japanese population, we examined the Arg110Gln variant using PCR-RFLP to compare the genotype and allele frequencies between 95 patients with cedar pollinosis and 95 healthy control subjects. Relationships between the Arg110Gln variant and the pollinosis-related traits, e.g. rhinitis severity, eosinophil counts in nasal secretion and serum total and allergen specific IgE levels, were also investigated. RESULTS: The frequencies of the minor allele Gln110 were 25.8% in patients with cedar pollinosis and 30.9% in healthy control subjects (p > 0.05). There was also no significant difference in the genotype frequencies between cases and controls (p > 0.05). In addition, we found no significant association of the Arg110Gln variant with any of the pollinosis-related phenotypes (p > 0.05). CONCLUSIONS: Our data suggest lack of evidence for identifying the variant Arg110Glnat the IL13 locus as a genetic risk factor involved in the development of Japanese cedar pollinosis. PMID- 16272824 TI - Sopoongsan inhibits mast cell-mediated anaphylactic reactions and inflammatory cytokine secretion. AB - BACKGROUND: Mast cells are key effector cells in the early-phase allergic inflammation and in diverse immunological and pathological processes. Sopoongsan (SPS), a traditional Korean medicine, has been used as therapeutics for allergic diseases such as atopic dermatitis (AD). The precise effect in experimental models of SPS, however, remains unknown. In this report, we investigated the effect of SPS on mast cell-mediated anaphylactic reactions and cytokine production in in vivo and in vitro murine models. METHODS: Compound 48/80-induced histamine and ear swelling were measured with the various concentrations of SPS. The amount of dye was determined colorimetrically after antidinitrophenyl IgE antibody-induced passive cutaneous anaphylaxis reaction. Secretion of tumor necrosis factor-alpha (TNF-alpha), interleukin-8 (IL-8) and IL-6 in supernatants from HMC-1 cells was measured by a sandwich enzyme-linked immunosorbent assay. The expression level of nuclear factor (NF)-kappaB/Rel A in the nucleus and the activation of mitogen-activated protein kinases (MAPKs) were examined by Western blot analysis. RESULTS: SPS inhibited the degranulation and histamine release from the rat peritoneal mast cells activated by compound 48/80. Compound 48/80 induced ear swelling was significantly reduced. SPS also showed an inhibitory effect of passive cutaneous anaphylaxis reaction. Significantly reduced levels (p < 0.05) of TNF-alpha, IL-8 and IL-6 were observed in the human mast cell line with SPS and SPS components. In addition, SPS inhibited an increase of NF-kappaB and extracellular signal-regulated kinase 1/2 activity. CONCLUSIONS: These findings suggest that SPS has an inhibitory effect on atopic allergic reaction and this might be useful for the clinical application to treat allergic diseases such as AD. PMID- 16272825 TI - Phosphatidylinositide 3-kinase is important in late-stage fibroblast growth factor-1-mediated angiogenesis in vivo. AB - We previously reported that overexpression of a secreted version of fibroblast growth factor-1 (sp-FGF-1) has the ability to induce angiogenesis in the chicken chorioallantoic membrane (CAM). In our current study, we examine the effects of sp-FGF-1 through a time course analysis of angiogenesis in the chicken CAM on days 3, 4, and 5 after gene transfection. Significant angiogenesis was observed on days 4 and 5 after gene transfection in the CAM assay. To evaluate the role of phosphatidylinositide 3-kinase (PI3K) signaling in sp-FGF-1-induced angiogenesis, we analyzed mRNA expression levels of PI3K and protein activity through its immediate downstream target, AKT-1. We found upregulation of both PI3K and AKT mRNA expression levels in day 5 sp-FGF-1 versus day 5 vector control-transfected CAMs. Furthermore, by blocking PI3K phosphorylation using the specific inhibitor, LY294002, we found that downstream phosphorylation of AKT-1 was inhibited. More importantly, the blockade of the PI3K pathway via LY294002 in sp-FGF-1 transfected CAMs significantly inhibited angiogenesis. These results further elucidate the molecular mechanisms of the sp-FGF-1 signaling pathway and it underscores the importance of PI3K signaling in FGF-1-stimulated angiogenesis in vivo. It also provides a basis for the role of sp-FGF-1 in the development of therapeutic treatments to combat vascular insufficiencies and angiogenesis dependent cancers. PMID- 16272826 TI - Critical appraisal and review of the Rankin scale and its derivatives. AB - BACKGROUND AND PURPOSE: Efforts to reduce stroke burden require accurate assessment of outcomes in order to compare treatments. The Rankin Scale and its derivatives, the Modified Rankin Scale and the Oxford Handicap Scale, taken together, are among the most common outcome measures that have been used in stroke research. The aim of this study was to perform a critical appraisal of the clinimetric properties of these scales. It was also planned to review the use of these scales in a selection of articles to illustrate concerns raised by the critical appraisal. SUMMARY OF REVIEW: A literature search was performed using electronic databases to locate relevant articles about the reviewed scales. The scales were appraised using a structured format regarding the following properties: purpose, development, presentation, language, method of administration, content validity, face validity, feasibility, construct validity, reliability, responsiveness, and generalizability. There are concerns in each of the appraised areas regarding the clinimetric properties of these scales. CONCLUSION: Further work is needed to improve the clinimetric properties of the reviewed scales to ensure that they are more useful tools in determining the outcome of stroke. Alternatively, a newer global outcome scale with improved clinimetric properties may be a better option for future stroke research. PMID- 16272827 TI - Psychological and physiological effects of 24-style taijiquan. AB - This study was conducted to determine whether 24-style Taijiquan (24TJQ) exhibits measurable psychological and physiological effects. Twenty-two middle-aged female subjects performed 24TJQ and cycle ergometry exercises at the same intensity determined by the same heart rate during 2 exercises. Electroencephalography and Profile of Mood State (POMS) were monitored before and after 2 exercises. The responses to 24TJQ exercise were different from those to cycle ergometry exercise when the heart rate returned to the resting level: (1) alpha increased and theta decreased significantly in the frontal region, while theta decreased significantly in the central and occipital regions; (2) in the POMS test, an improved positive mood was found following 2 exercises, while negative mood was suppressed following 24TJQ exercise; (3) significant correlations were found between the increased alpha in the frontal region, the decreased theta in the central or occipital region and the vigor of the POMS test. These results suggest that 24TJQ exercise induced a resting awakening state and exhibited a relaxing effect on both mind and body. PMID- 16272828 TI - Transcranial magnetic stimulation in clinical practice. AB - Transcranial magnetic stimulation allows a non-invasive and painless stimulation of the human brain and cranial nerves. The method is in use since 1985. Transcranial magnetic stimulation can use single stimuli, pairs of stimuli separated by different intervals (to the same or to several brain areas), or trains of repetitive stimuli at various frequencies. Single stimuli give rise to motor evoked potentials that have clinical use and serve diagnostic and prognostic purposes. Repetitive transcranial magnetic stimulation can modify excitability of cerebral cortex. Repetitive transcranial magnetic stimulation has opened a new field of investigation of the neural circuitry, and is developing into a therapeutic tool. This general review considers basic principles of transcranial magnetic stimulation, discusses methodological aspects and techniques, and analyses their utility in clinical practice. PMID- 16272829 TI - [Relationship between the metabolic syndrome, endothelial function and intima media thickness in asymptomatic middle-aged individuals]. AB - OBJECTIVE: To evaluate the relation between cardiovascular risk factors, metabolic syndrome, endothelial function and carotid intima-media thickness in asymptomatic middle-aged individuals. MATERIALS AND METHODS: A total of 180 subjects (86 men and 94 women, mean age 38.8+/-0.3 years) have been investigated. Anthropometric, blood pressure measurements have been performed and lipid as well as high sensitivity C-reactive protein blood levels have been tested. The endothelial function was assessed by measuring the vasodilatation of the brachial artery. The carotid intima-media thickness was measured with high resolution B mode ultrasound imaging. Metabolic syndrome was diagnosed using International Diabetes Federation definition criteria (2005). RESULTS: The metabolic syndrome has been diagnosed in 48 (26.7%) individuals: 28 (32.6%) males and 20 (21.3%) females. Significantly reduced endothelial function has been established in asymptomatic men when compared to women. However, statistically significant decrease (4.84%) in endothelial function in metabolic individuals was detected only among males. In the metabolic patients intima-media of common carotid artery, carotid bulb and internal carotid artery was thicker than in those without metabolic syndrome (0.006, 0.007 and 0.007 cm, respectively) (alpha=0.000, beta=0.01). Endothelial dysfunction and intima-media thickening correlated with increased blood pressure, abdominal circumference and body mass index as well as with elevated blood triglyceride and glucose levels. Intima media thickness was greater in individuals with impaired endothelial function in all carotid segments tested. No relationship has been observed between total or low-density lipoprotein cholesterol concentrations and endothelial function, intima-media thickening or high sensitivity C-reactive protein levels. CONCLUSIONS: These observations suggest that the metabolic syndrome and/or its components may influence the different initial mechanisms of atherosclerosis- disorder of endothelial function and intima-media thickening. It is presumable that the lipid disorders as well as inflammation may play more significant role in the presence of impaired endothelial function. PMID- 16272830 TI - Gastrointestinal disorders in children with atopic dermatitis. AB - The aims of this study were to analyze the peculiarities of allergies to food; to determine gastrointestinal disorders, endoscopic signs of mucosal damage and histological lesions of the mucosa and to establish their relation to the extent of atopic dermatitis and its degree of severity. MATERIAL AND METHODS: A total of 164 children (86 boys and 78 girls) suffering only from atopic dermatitis were examined. Atopic dermatitis was diagnosed using standard diagnostic criteria; extent of disease (the Basic Clinical Scoring System (BCSS)) and the severity (Scoring Atopic Dermatitis (SCORAD) index), total serum IgE levels were determined; skin prick and patch tests with the main food allergens were performed. Using questionnaire gastrointestinal disorders with the symptoms of atopic dermatitis were ascertained. In children with atopic dermatitis suffering from chronic dyspepsia esophagealgastroduodenoscopy was performed and biopsy samples from the antrum of the stomach and duodenum were taken. RESULTS: The age of patients ranged from 6 months to 18 years. According to extent of atopic dermatitis and degree of severity localized, mild atopic dermatitis prevailed. Analysis of the changes in total Ig E levels showed different degree of sensitization of the children examined. Considering the type of allergic reaction, immediate-type allergic reactions dominated only in 11.6% of children with atopic dermatitis, whereas delayed-type allergic reactions manifested in 44.5% of children. No food allergy was present in one-fifth of children with atopic dermatitis. One hundred four (63.4%) children complained of gastrointestinal disorders. Of these 104 patients, 17 children (mean age 6.9 years) who underwent esophagealgastroduodenoscopy with biopsy had no pathology; however, histological examination of mucosa revealed eosinophilic infiltration in the gastric antrum and duodenum in three children. CONCLUSIONS: The most common gastrointestinal disorders are: abdominal pain vomiting, diarrhea, abdominal distention, and constipation. The incidence of gastrointestinal disorders does not depend on the extent and severity of atopic dermatitis. Gastrointestinal disorders manifest independent of the type of allergic reactions inducing atopic dermatitis. The most common food allergens, such as soy, milk, peanuts, corn, carrots, rye, wheat, white of the egg, cod, and chicken were determined by skin patch test. According to our data, no pathology was found in children with atopic dermatitis during endoscopic examination. Our data showed that allergic-like changes in gastric and duodenal mucosa were present only in older (7-10-year-old) children. PMID- 16272831 TI - Analysis of burned hand function (early versus delayed treatment). AB - The aim of this study was analyze the results of hand function in adult patients with deep partial thickness hand burns. All patients were treated in Kaunas University of Medicine Hospital in Lithuania during the period 2001-2004. MATERIAL AND METHODS: A total of 79 cases were reviewed prospectively. We divided the studied patients into two groups by the envelope method (A group of early and B of delayed necrectomy and plasty). Two treatment methods--early and delayed surgery--have been analyzed. We present results of analysis of hand function in the periods of 3 and 12 months after the burn injuries of the hands. RESULTS: During the study we ascertain that the deficiency of hand function in group A 3 and 12 months after the injury was lower than that in group B. The deficiencies of hand and arm functions decreased over time. After a period of 12 months the deficiency of hand and arm functions in group A reached 12%, while in group B the deficiencies of hand and arm function reached 23% and 26.5%, respectively. The study revealed that the deficiency of hand function because of wrist and hand burn depended on the age of patient, the area of hand burn, time period from the injury to the arriving to a hospital and time to the operation. The strength of digit pinch and hand grasp was larger in group A. Twelve months after the burn the strength measured in A and B groups was larger than that measured 3 months after the burn. The strength of hand grasp in group A in male patients regenerated up to 76%, in female--61% of norm. In group B the strength of hand grasp in male patients regenerated up to 60.8% and in female--39.36%. CONCLUSIONS: During the perspective analysis it was determined that after periods of 3 and 12 months after the injury the strength pinch and grip was statistically significantly larger in group A. The deficiency of hand function, which indicates the general hand function, was statistically significantly lower in group A. PMID- 16272832 TI - Occupational therapy for patients with spinal cord injury in early rehabilitation. AB - OBJECTIVE: To determine factors influencing the effectiveness of occupational therapy of patients with spinal cord injury in early rehabilitation. MATERIAL AND METHODS: Data were obtained on 136 patients with spinal cord injury admitted to the Department of Rehabilitation, Kaunas University of Medicine Hospital in 1999 2005. The study population consisted of 97 (71.3%) males and 39 (28.7%) females. Complex rehabilitation was started after the stabilization of functional state in the Department of Neurosurgery and transfer to Department of Rehabilitation. The average duration of early rehabilitation in the Department of Rehabilitation was 68.3+/-22.5 days. According to the level of spinal cord injury patients were divided into two groups: patients with cervical lesions (C1-Th1 segments) and with thoracic-lumbar lesions (Th2-S1 segments). According to the level of spinal cord injury patients also were divided into two groups: complete (American Spinal Injury Association (ASIA-A)) and incomplete injury (ASIA-B, ASIA-C). The evaluation of the level of patients' independence was dependent on the level of injury: C4; C5; C6; C7-C8; Th1-Th9; Th10-L1; L2-S5 segments. Functional state and activity were evaluated by Functional Independence Measure (FIM). Effectiveness of occupational therapy was supposed to be good, if predicted independence level of final rehabilitation was achieved by a patient and it was supposed to be unsatisfactory if the same level of independence was not achieved. RESULTS: The examination of patients has shown that 21 (15.4%) patients had complete injury (ASIA-A) in cervical level and 41 (30.2%) patients had complete injury in thoracic-lumbar level. Thirty-five (25.7%) patients had incomplete injury in cervical area and 39 (28.7%) patients in thoracic-lumbar level. At the end of early rehabilitation period in the case of complete spinal cord injury in cervical level (C4-C8) the expected level was achieved by 33.3-100% of patients. In the case of incomplete injury the expected level was achieved by 87.5-100% of patients. No patient with spinal cord injury in thoracic-lumbar (Th1-S5) level achieved the expected level of independence of final rehabilitation in early rehabilitation period. CONCLUSIONS: Patients with spinal cord injury had disturbances of all activities according to FIM in early rehabilitation. The level of spinal cord injury had greater influence on the level of independence of patients with injury in cervical level rather than in thoracic-lumbar level. The level of independence in early rehabilitation period was achieved in 17.7% of complete spinal cord injury cases and in 45.9% of incomplete spinal cord injury cases. PMID- 16272834 TI - Extensive limb lengthening in Ollier's disease: 25-year follow-up. AB - A case of extensive lower limb lengthening (32 cm) in a 14-year-old male patient with Ollier's disease is reported. A varus deformity of the femur and a valgus deformity of the tibia were evident. The femur was successfully lengthened 22 cm by metaphyseal distraction, and the tibia was lengthened 10 cm by two-stage distraction-compression method with a cylindrical bone allograft. Ilizarov's distraction device was used. Radiologically, a good bone regenerate was formed. Host bone has incorporated (like sarcophagi) the allograft of tibia. No evidence of vascular or neural disturbances was found. The lengthening indices were counted for femur 22.5 days per centimeter and for tibia 21 days per centimeter, altogether 15.5 days per centimeter. Bone lengthening was performed through the Ollier's disease foci. Fine needle biopsy investigation showed that most embryonic cartilage cells had been replaced with bone tissue. After five years and a 25-year follow-up the patient was satisfied with the result. The function of the knee joint was limited, but the limb was fully weight-bearing. Signs of knee osteoarthritis were found. PMID- 16272833 TI - [Successful treatment with continuous infusion of coagulation factor concentrates in severe hemophilia A complicated with mixed trauma, crush syndrome and acute renal failure]. AB - Treatment method of hemophilia with continuous infusion of clotting factor concentrates is not widespread. We report our experience with the use of continuous infusion of factor VIII in patient with mixed trauma, crush syndrome and acute renal failure. Patient was successfully treated with fasciotomy of left upper arm and forearm, 6 procedures of hemodialysis, 34.500 units of clotting factor VIII concentrate, and 13 red blood cells units. Our experience confirms the safety, efficiency, and economy of the method of the constant infusion of clotting factor concentrates for hemophilia patient requiring replacement therapy, surgical intervention, and hemodialysis. PMID- 16272835 TI - [The prevalence of metabolic syndrome in middle-aged in Kaunas population]. AB - AIM: To assess the prevalence of the metabolic syndrome among middle-aged Kaunas population. MATERIAL AND METHODS: A total of 433 (192 males and 241 females, mean age 38.8+/-0.3 years) asymptomatic individuals have been investigated. Metabolic syndrome was diagnosed using International Diabetes Federation definition criteria - central obesity (the waist circumference > or =94 cm for men and > or =80 cm for women) plus any two of the following four factors: elevated triglycerides (Tg) (> or =1.7 mmol/L), low high-density lipoprotein cholesterol (HDL-C) levels (<1.0 mmol/L for men and <1.3 mmol/L for women), blood pressure (BP) of 130/85 or higher (systolic pressure >130 mmHg or a diastolic pressure >85 mmHg) or hyperglycemia (> or =5.6 mmol/L). RESULTS: The metabolic syndrome was present in 21.7% (28.1% among males and 16.6% among females) of individuals. Furthermore, 41.1% had an increased waist circumference, 56.4% had elevated BP (130/85 or higher), 31.2% had elevated plasma glucose concentration, 19.9% had low HDL-C cholesterol values and 14.1% had increased Tg values. Significantly higher concentration (1.16 mg/L, alpha<0,001, beta<0.001) of high-sensitivity C reactive protein has been established in individuals with the metabolic syndrome. An increased concentration (>3 mg/L) of high-sensitivity C-reactive protein was more prevalent (17.3%) in the cohort with metabolic syndrome. Concentration of high-sensitivity C-reactive protein directly correlated with the waist and hips circumference, body mass index, concentration of Tg, glucose and BP. CONCLUSIONS: The prevalence of the metabolic syndrome among middle-aged Kaunas residents was 22%. Metabolic syndrome and central obesity correlated with elevated concentration of high-sensitivity C-reactive protein. Moderately (1-3 mg/L) and severely (>3 mg/L) elevated high-sensitivity C-reactive protein levels increase the chance of metabolic syndrome by 4.15 and 7.39 times, respectively (p=0,000). An innovative approach towards cardiovascular risk assessment integrating traditional cardiovascular risk factors, metabolic syndrome and high-sensitivity C-reactive protein values could improve the risk stratification in asymptomatic middle-aged population. PMID- 16272836 TI - [The economic estimates of well-timed diagnostics and early treatment of type 2 diabetes mellitus]. AB - Type 2 diabetes mellitus is a severe, high-priced, and to date not curable, however, controllable disease. When diabetes mellitus is not treated or is poorly controlled, complications of small and large blood vessels arise, and the mortality increases. The costs of treatment of diabetes-related complications are very high. Estimates performed in other countries show that expenses on treatment of diabetes-related complications are considerably higher than the expenses paid for diabetes control itself. Type 2 diabetes mellitus is often diagnosed too late, when complications requiring expensive treatment are already present. OBJECTIVE: The aim of the study was to estimate the impact of well-timed diagnostics and early treatment of type 2 diabetes mellitus on total health care costs. METHODS: Direct and indirect costs of treatment of the disease and its complications were estimated using Markov model with a discount of 5% per annum. RESULTS: It was calculated that with well-timed diagnostics and early initiation of treatment of type 2 diabetes mellitus the total costs to budget of the Compulsory Health Insurance Fund per one patient would be reduced by 1,736 Lt over 5 years and over 10 years our state would save 5,911 Lt. Not only direct but indirect costs would be reduced as well. The costs of 2,418 Lt would be saved per one patient over 5 years, and 7,061 Lt - over 10 years. With improvement of diagnostics of type 2 diabetes mellitus and with well-timed diagnostics in at least 50% of cases of diabetes mellitus in Lithuania, the health budget could save 34.7 million Lt over 5 years, and even 118.2 million Lt over 10 years. Diagnosis of type 2 diabetes mellitus made in time and administration of reimbursed drugs would prolong the life of a patient with type 2 diabetes mellitus by 2.67 months over 10 years. CONCLUSION: Diagnosis of type 2 diabetes mellitus in time provides a possibility to reduce expenses of diabetes treatment and care and to prolong a patient's life. PMID- 16272837 TI - [Peculiarities of problem-based learning in medical education]. AB - In most Western universities at the end of the 20th century and at the beginning of the 21st problem-based learning is treated as a new direction of didactics or as challenge to traditional study system. Some Western researchers name problem based learning as the method which most corresponds to the goals of modern medical education. When implementing the system of problem-based learning into traditional study systems, the junction problems can be conditioned by work principles of teacher performing tutor's functions and a traditional one. Student oriented teaching conditions the junction between traditional approach, which emphasizes student's dependence on a teacher as information source and modern one stressing student's freedom when choosing the values of education and responsibility for his/her learning. On integrating the content of various subjects, junction problems are conditioned by disappearance of classical borders between separate subjects. When emphasizing the solution of problems in study programs, transition from the teaching concentrated on separate subject studies to the studies based on systemic problem solution can cause junction problems. Realizing the study program content oriented to community, the junction can be conditioned by insufficient openness of university to society, as, first of all, one has to train not only students and teachers but also society for new interaction with university as an institution that realizes health study program. The objective of the article is to reveal the peculiarities of problem-based learning when realizing the aims of medical education. PMID- 16272838 TI - [Electrical defibrillation and cardioversion]. AB - Defibrillation and cardioversion are techniques in which a short electric impulse is administered to the heart in order to restore its normal rhythm. During cardioversion electric impulse is synchronized to the QRS on electrocardiogram. During defibrillation electric current passes through the heart in any phase of electric heart cycle. This mode of treatment is rather new - Lown et al. started to use it in a clinical practice in 1962. During defibrillation or cardioversion electric current goes from negative to positive electrode of defibrillator and passes the heart on its way. This induces transmembrane potential in myocardium cells and results in synchronic depolarization of all myocardium. The pathophysiology of defibrillation is explained by critical mass hypothesis as well as the upper limit of vulnerability hypothesis. The success of defibrillation depends on many factors, such as the location and size of electrodes, the type of defibrillator, the morphology of electric impulse, transthoracic impedance, the type and duration of arrhythmia. This procedure can be performed only on unconscious patient. The possible complications of the procedure can be disturbances in heart rhythm and conduction, the changes in arterial blood pressure, the damage to the myocardium, embolia, pulmonary edema and others. This article describes the mechanism of action of defibrillation and cardioversion, indications for this procedure, the technique and methods of defibrillation and cardioversion, the factors, responsible for the efficacy of the procedure and possible complications of defibrillation. PMID- 16272839 TI - Three-dimensional computed tomographic imaging of complex congenital cardiovascular abnormalities. AB - Primary evaluation of congenital cardiac abnormalities traditionally relies upon echocardiography and conventional angiography, both of which have potential limitations. Echocardiography is an operator dependant study, limited by a small window and patient movement. Conventional angiography is an invasive procedure with an inherent risk of catheter complication such as vessel damage, bleeding, stroke and infection. During angiography, overlapping of the pulmonary and systemic circulation often provides a confusing picture given complex anatomy. Another limiting factor of particular significance in young children is radiation dose and contrast administration during catherization procedures. Three dimensional MDCT provides an alternative to alleviate these pitfalls of traditional cardiac diagnostic studies. Development of multi-detector computed tomography (MDCT) and 3D software provides new methods for non-invasive visualization and evaluation of congenital cardiac abnormalities. The multiplanar, volumetric functions allow faster and more complete computed tomography diagnosis and better understanding of clinical relevance of complex cardiac anatomy. In addition, 3D imaging is particularly useful for preoperative planning and postoperative outcomes. This essay provides case studies to illustrate the usefulness of MDCT/3D CT for evaluation of complex congenital heart disease. PMID- 16272840 TI - Right ventricle shape and contraction patterns and relation to magnetic resonance imaging findings. AB - OBJECTIVE: To analyze and to describe the shape and contraction of the normal right ventricle (RV) as visualized by magnetic resonance imaging (MRI). METHODS: Thirty normal volunteers were imaged using cine MRI in axial, short-axis, and long-axis planes. The shape and contraction of the RV were qualitatively evaluated. Quantitative evaluation of RV shape was performed by calculating the angle subtended between the planes of horizontal long-axis view (HLA) and axial view and the RV base-to-apex distance. Multiplanar reformation was used to visualize changes between corresponding views. RESULTS: The spectrum of major RV shape (wedge, box, and round) was more variable on axial images (17%, 43%, and 23%, respectively) than on HLA images (63%, 20%, and 0%, respectively). Focal outpouching of the RV free wall was more frequent on the axial view than on the HLA view. The subtended plane angle and base-to-apex distance showed statistically significant dependence indicative of an artificially foreshortened RV in the axial view with a direct influence on RV variations. CONCLUSIONS: With increasing subtended angles, variation of the normal RV appearance is substantially higher on axial views compared with HLA views. PMID- 16272842 TI - Comprehensive multidetector computed tomography assessment of severe cardiac contusion in a pediatric patient: correlation with echocardiography. AB - Multidetector computed tomography (MDCT) cardiac findings in an unconscious teenager after blunt chest trauma are presented. Multidetector computed tomography enabled accurate comprehensive evaluation of the coronary arteries, myocardial perfusion, and left ventricular function. This case illustrates the full capabilities of MDCT in the evaluation of cardiac contusion in a noncooperative pediatric patient. PMID- 16272841 TI - Image quality and diagnostic accuracy of 16-slice multidetector spiral computed tomography for the detection of coronary artery disease in elderly patients. AB - OBJECTIVES: To determine the image quality and diagnostic accuracy of cardiac multislice spiral computed tomography (MSCT) in elderly patients (>65 years old) in comparison to younger patients, this retrospective analysis was performed. METHODS: The catheter-controlled MSCT results from patients older than 65 years of age were compared with the results of younger patients in a cohort of 117 patients with regard to sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and image quality. Fifty-three patients were older than 65 years of age (group 1: 31 men, age range: 72.2 +/- 4.1 years, number of risk factors: 2.6 +/- 1.3, Agatston score: 866 +/- 1090) and 64 were younger (group 2: 45 men, age range: 57.4 +/- 6.1 years, number of risk factors: 2.6 +/- 1.0, Agatston score: 765 +/- 1013). All patients were examined by MSCT (Sensation 16 Speed 4 D; Siemens, Forchheim, Germany, with a gantry rotation time of 375 milliseconds) and invasive coronary angiography. The MSCT results were compared blinded with the results of the coronary angiography with regard to the presence or absence of significant stenosis (>50%) in a 13-segment model. Image quality was assessed on a qualitative scale between 1 (very good) and 5 (insufficient image quality) for each segment. RESULTS: Sensitivity, specificity, PPV, and NPP were not different statistically in both groups (group 1: 0.80/0.96/0.89/0.93 and group 2: 0.89/0.98/0.93/0.97). Three patients (all <65 years old) had to be excluded from analysis because of technical problems. Image quality was significantly better in group 2. Gender, body mass index, number of risk factors, and mean heart rate were not significantly different in either group. CONCLUSIONS: Age has an impact on MSCT image quality but did not hamper diagnostic accuracy. Thus, MSCT is a noninvasive method to detect or rule out coronary artery disease independently of age. These retrospective data have to be confirmed in larger prospective trials. PMID- 16272843 TI - Giant cell myocarditis depicted by cardiac magnetic resonance imaging. AB - Giant cell myocarditis is a rare condition, the cardiac magnetic resonance imaging findings of which have not been previously described. The disease usually occurs in young previously healthy people and is typified by rapidly progressive cardiac dysfunction, often requiring cardiac transplantation. A case of giant cell myocarditis is presented, with associated pathologic and imaging findings. PMID- 16272844 TI - Demonstration of cardiac involvement of sarcoidosis by contrast-enhanced multislice computed tomography and delayed-enhanced magnetic resonance imaging. AB - Delayed-enhanced magnetic resonance (MR) imaging has recently been shown to be effective in detecting cardiac sarcoidosis. Two cases in which contrast-enhanced multislice computed tomography imaging clearly identified the localization and extension of cardiac sarcoidosis as delayed-enhanced MR imaging are presented. PMID- 16272846 TI - Evaluation of unroofing procedure of anomalous origin of right coronary artery from left sinus of valsalva between aorta and pulmonary trunk by multidetector computed tomography. AB - An anomalous origin of a coronary artery is a rare congenital anomaly. Conventionally, the diagnosis of coronary artery anomalies is performed by coronary angiography. Currently developed multidetector computed tomography has permitted better definition of the coronary arteries, however. Multidetector computed tomography could be used effectively in the diagnosis and evaluation of the postoperative status of a patient with an anomalous origin of the right coronary artery from the left sinus of Valsalva between the aorta and pulmonary trunk. PMID- 16272845 TI - Cardiac pseudosarcomatous fibromyxoid tumor: a review of the literature. AB - A cardiac pseudosarcomatous fibromyxoid tumor (PFT) is described in a previously healthy 35-year-old man, together with a review of the literature. Pseudosarcomatous fibromyxoid tumor is within the spectrum of inflammatory (myofibroblastic) pseudotumors. It has previously been described predominantly within the genitourinary tract and respiratory tract. Inflammatory pseudotumor is rare as a cardiac tumor, and cardiac PFT is not previously reported. No recurrence or metastatic disease has been reported after resection of PFTs elsewhere in the body, and this tumor seems to have a benign clinical course. PMID- 16272847 TI - Magnetic resonance imaging diagnosis of carcinoid heart disease. AB - Carcinoid tumors are associated with the production of a number of bioactive substances that might deposit on the endocardium (endocardial fibroelastosis) and produce cardiac failure. Carcinoid heart disease usually presents as right-sided valvular heart disease. Even if the diagnosis is frequently made by transthoracic echocardiography, cardiac magnetic resonance imaging adds valuable information when evaluating cardiac structures that might be difficult to analyze on echocardiography. PMID- 16272848 TI - Preliminary report of contrast-enhanced computed tomography for patients with a percutaneous cardiopulmonary support system. AB - The purpose of this study was to investigate a suitable protocol of contrast enhanced computed tomography (CECT) in cases with a cardiopulmonary support system. Contrast-enhanced computed tomography with intra-arterial injection (IAI) of contrast medium (CM) via a perfusion cannula showed sufficient contrast enhancement in 2 cases of cardiac decompensation (CD). Contrast-enhanced computed tomography with intravenous injection of CM showed insufficient and delayed contrast enhancement of the aorta in 2 cases of CD and 3 cases of pulseless electrical activity. We encourage administration of CM by means of IAI. PMID- 16272849 TI - Eosinophilic hepatic necrosis: magnetic resonance imaging and computed tomography comparison. AB - OBJECTIVE: To compare the findings of magnetic resonance (MR) imaging with those of computed tomography (CT) of focal liver lesions related to peripheral eosinophilia. METHODS: For 12 patients with peripheral eosinophilia (>7%) examined with hepatic MR imaging and CT, 52 focal hepatic lesions larger than 0.5 cm, including 31 lesions simultaneously found on the 2 imaging modalities, were subjected to a comparative analysis of their imaging features. RESULTS: The total number of lesions distinguished from background liver was 39 (75%) on MR imaging and 44 (85%) on CT scans. On arterial phase images of 10 patients with comparable data, homogeneously hyperintense lesions were demonstrated more frequently (P = 0.006) on MR imaging (16 [50%] of 32 lesions) than on CT scans (4 [13%] of 32 lesions). Only 7 (22%) of the 32 hypoattenuating lesions on portal phase CT were depicted as hypointense lesions on portal phase MR images in 12 patients. On delayed phase images in 8 patients, the number of hyperintense lesions on MR images (9 [56%] of 16) was greater (P = 0.077) than that seen on the CT scans (4 [25%] of 16). CONCLUSIONS: For many focal hepatic lesions related to peripheral eosinophilia, dynamic MR imaging more easily demonstrates lesional enhancement on arterial and delayed phases than CT scans. Because of the higher degree of lesional enhancement of MR imaging compared with CT, the lesion-to-liver contrast may not be sufficient to distinguish the lesion from the background liver, resulting in decreased sensitivity of portal phase dynamic MR imaging. PMID- 16272850 TI - Benign or malignant?: differentiating breast lesions with computed tomography attenuation values on dynamic computed tomography mammography. AB - OBJECTIVE: To evaluate enhancement effects of breast lesions with dynamic computed tomography (CT) and to determine characteristics to aid in differential diagnosis of benign and malignant lesions. METHODS: One hundred seventy-six women underwent preoperative dynamic CT, in which they were scanned with rapid injection of contrast media (3 mL/s) after 30 seconds and 2 minutes (early and delayed phases, respectively). The CT values and enhancement patterns of malignant lesions (n = 154) were compared with those of benign lesions (n = 22), and the cut point of CT values with the best validity was analyzed. RESULTS: The CT values of malignant lesions were higher than those of benign lesions in both phases (P < 0.0001). The cut point was determined to be 60 Hounsfield units (HU) in the early phase (44% validity, 90% sensitivity). Washout and plateau patterns were more commonly seen in the malignant group (73% specificity). CONCLUSIONS: The analysis of enhancement effects on CT may lead to more appropriate differentiation of benign and malignant lesions. PMID- 16272851 TI - Usefulness of a saline flush for intravenous 3-dimensional computed tomography portography using multidetector-row helical computed tomography. AB - PURPOSE: To evaluate the clinical usefulness of a saline flush technique in improving the imaging quality of 3-dimensional computed tomography portography (3D-CTP). METHODS: To evaluate liver metastases, 58 patients were divided into 2 groups undergoing 3D-CTP with or without a saline flush. The computed tomography (CT) values of the right portal vein (RPV), left portal vein (LPV), main portal vein (MPV), and right hepatic parenchyma (RHP) were assessed. Maximum intensity projection (MIP) 3D-CTP images were evaluated by vessel visualization. RESULTS: Higher mean CT attenuation values in the RPV, LPV, MPV, and RPV-RHP were observed in the saline flush group and were statistically significant (P = 0.04, P = 0.03, P = 0.01, and P = 0.04, respectively). The difference in imaging quality between 2 groups was statistically significant (P = 0.04). In segment VIII, the ability to depict the segmental branches was significantly higher when the saline flush technique was used (P = 0.03). CONCLUSIONS: The saline flush technique increases the CT attenuation values of the portal vein and the difference in values between the portal vein and the tissue around it and improves the MIP imaging quality of 3D-CTP. PMID- 16272852 TI - Benign mixed epithelial and stromal tumor of the kidney: imaging findings. AB - Three cases of mixed epithelial and stromal tumor of the kidney with their imaging findings are described; these cases have not been reported previously in the radiology literature. This benign tumor contains epithelial and spindle cell stromal components and arises exclusively in adult women. It is characterized morphologically by a solid and cystic nature with delayed enhancement. PMID- 16272853 TI - Bilateral scrotal extraperitoneal herniation of ureters: computed tomography urographic findings and review of the literature. AB - Scrotal herniation of the ureter is an extremely rare and often misdiagnosed condition with possible serious surgical complications. Most reported cases occur in obese men during the fifth and sixth decades of life. Two types of ureteroinguinal hernias are defined: paraperitoneal (more frequent and acquired form with a peritoneal hernia sac in which other abdominal structures are found) and extraperitoneal (uncommon and congenital form without a peritoneal sac that is almost always composed of the ureter and fat tissue). A case of scrotal extraperitoneal ureteral hernia with exquisite computed tomography urographic and ultrasound findings is presented. PMID- 16272854 TI - Pathologic subgroups of nonspecific interstitial pneumonia: differential diagnosis from other idiopathic interstitial pneumonias on high-resolution computed tomography. AB - OBJECTIVE: To determine whether the subtypes of nonspecific interstitial pneumonia (NSIP) could be differentiated from other idiopathic interstitial pneumonias (IIPs) on the basis of findings on high-resolution computed tomography (CT). METHODS: Two observers evaluated the high-resolution CT findings in 90 patients with IIPs. The patients included 36 with NSIP, 11 with usual interstitial pneumonia (UIP), 8 with cryptogenic organizing pneumonia (COP), 10 with acute interstitial pneumonia (AIP), 14 with desquamative interstitial pneumonia (DIP) or respiratory bronchiolitis-associated interstitial lung disease (RB-ILD), and 11 with lymphoid interstitial pneumonia (LIP). The NSIP cases were subdivided into group 1 NSIP (n = 6), group 2 NSIP (n = 15), and group 3 NSIP (n = 15). RESULTS: Observers made a correct diagnosis with a high level of confidence in 65% of NSIP cases, 91% of UIP cases, 44% of COP cases, 40% of AIP cases, 32% of DIP or RB-ILD cases, and 82% of LIP cases. Group 1 NSIP was misdiagnosed as AIP, DIP or RB-ILD, and LIP in 8.3% of patients, respectively. Group 2 NSIP was misdiagnosed as COP in 10% of patients, LIP in 6.7%, AIP in 3.3%, and DIP or RB-ILD in 3.3%. Group 3 NSIP was misdiagnosed as UIP in 6.7% of patients, COP in 6.7%, and DIP or RB-ILD in 3.3%. CONCLUSIONS: In most patients, NSIP can be distinguished from other IIPs based on the findings on high resolution CT. Only a small percentage of patients with predominantly fibrotic NSIP (group 3 NSIP) show overlap with the high-resolution CT findings of UIP. PMID- 16272855 TI - Specific high-resolution computed tomography findings associated with sputum smear-positive pulmonary tuberculosis. AB - OBJECTIVES: This study aimed to evaluate whether high-resolution computed tomography (HRCT) could predict the results of a sputum smear in patients with active pulmonary tuberculosis. METHODS: Forty-eight patients with active pulmonary tuberculosis were divided into 2 groups: sputum smear-positive (n = 25) and -negative (n = 23). The HRCT findings were retrospectively reviewed, focusing on the presence or absence of features previously reported to indicate active pulmonary tuberculosis, including ground-glass opacity, cavitation, centrilobular opacity, and air space consolidation. RESULTS: Although air space consolidation was the least common feature overall, it occurred significantly more frequently in the smear-positive group than in the smear-negative group. This feature also had the highest specificity and positive predictive value. Cavitation and ground glass opacity also occurred significantly more frequently in the smear-positive group. The frequency of centrilobular opacity did not differ between the 2 groups. CONCLUSIONS: The present study suggested that the HRCT findings of air space consolidation, cavitation, and ground-glass opacity are significantly associated with smear-positive pulmonary tuberculosis. PMID- 16272856 TI - Sixteen-slice multidetector computed tomography pulmonary angiography: evaluation of cardiogenic motion artifacts and influence of rotation time on image quality. AB - OBJECTIVE: To describe the site and severity of cardiogenic motion artifacts on 16-slice multidetector computed tomography (CT) angiograms of the pulmonary circulation and to determine the impact of the gantry rotation time on image quality. METHODS: Sixty-nine patients underwent nonelectrocardiographically gated 16-slice multidetector CT angiography of the pulmonary circulation with a rotation time of 0.5 second (group 1, n = 37) or 0.375 second (group 2, n = 32). All scans were prospectively analyzed for the presence of cardiogenic motion artifacts in the pulmonary arteries, defined as a blurring of the edges of the pulmonary arteries on lung window settings with or without an artifactual loss of enhancement in the arterial lumen on soft tissue windows ("pseudofilling defect"). In addition, the severity of pulmonary arterial blurring was recorded using a 4-point scale. RESULTS: Fifty-six patients (81%) had at least 1 pulmonary artery affected by cardiogenic motion artifacts. At the level of the central pulmonary arteries, movement artifacts responsible for a blurring of the arterial wall were present in 35 patients (51%), mainly observed at the level of the pulmonary trunk (n = 28, 41% of patients) and right main pulmonary artery (n = 23, 33% of patients); in 2 patients, motion artifact was responsible for a pseudoflap appearance within the arterial lumen (n = 2, 4% of patients). Motion artifacts at the level of lobar pulmonary arteries were noted only in the lower divisions of the lingula (n = 1, 1.5% of patients) and in the left lower lobe (n = 2, 3% of patients). At the level of the segmental pulmonary arteries, motion artifacts were most frequently found in the lower divisions of the left upper lobe (lingula) (n = 19, 27.5% of patients) and left lower lobe (n = 29, 42% of patients). At the subsegmental level, motion artifacts were most common in the lower divisions of the left upper lobe (lingula) (n = 30, 43.5% of patients) and left lower lobe (n = 35, 51% of patients). Pseudofilling defects were depicted at the segmental and subsegmental levels in 28 patients (41% of the study group), almost exclusively located in the lingula and left lower lobe. Cardiogenic motion artifacts were observed with a significantly lower frequency in group 2 (22 of 32 patients, 69%) than in group 1 (34 of 37 patients, 92%) (P = 0.0142), with a concurrent reduction in the frequency of pseudofilling defects identified in 20 patients (54%) in group 1 and in 8 patients (25%) in group 2 (P = 0.0142). CONCLUSION: The use of a 0.375-second rotation time enables significant reduction in the frequency of cardiogenic motion artifacts on 16-slice multidetector CT angiograms of the pulmonary circulation. PMID- 16272857 TI - Computed tomography findings of pathologically confirmed pulmonary parenchymal endometriosis. AB - OBJECTIVE: To evaluate the usefulness of computed tomography (CT) in the localization of parenchymal pulmonary endometriosis and to correlate the CT findings with fiberoptic bronchoscopic and pathologic findings. METHODS: A prospective study of 5 patients presenting with catamenial hemoptysis was conducted. The CT scans and fiberoptic bronchoscopy were performed twice during and 2 weeks after menstruation. After the localization of the presumed bleeding focus, surgical resection was performed. RESULTS: The CT scans obtained during menstruation revealed a well-demarcated area of consolidation (n = 4) and ground glass opacity (n = 5), whereas CT scans obtained after menstruation demonstrated ground-glass opacity (n = 4) or complete resolution of the previously noted lesion (n = 1). Fiberoptic bronchoscopy exhibited trails of blood clot at the orifice of the involved bronchi unilaterally (n = 4) or a thin bloody secretion in the bronchi bilaterally. Histopathologic examination of the resected specimens showed typical findings of pulmonary endometriosis. CONCLUSION: Computed tomography scans during and after menstruation were useful for the precise preoperative localization of parenchymal pulmonary endometriosis. PMID- 16272858 TI - Reliability of quantitative computed tomography to predict postoperative lung function in patients with chronic obstructive pulmonary disease having a lobectomy. AB - OBJECTIVE: To verify the reliability of quantitative computed tomography (CT) to estimate the postoperative lung function in patients with mild to severe chronic obstructive pulmonary disease (COPD) who underwent a lobectomy. METHODS: Nine COPD patients with lung cancer having a lung lobectomy with preoperative CT were enrolled. By applying a density mask technique and a specific equation, predicted postoperative forced expiratory volume in 1 second (FEV1) and vital capacity (VC) were calculated. Predicted values were correlated with postoperative measured values. RESULTS: Estimated FEV1 and VC were always significantly lower than the corresponding postoperative values; however, CT-estimated postresection FEV1 values were better than the postresection VC values (biases between estimated and measured values were -0.14 and -0.536 L, respectively, according to the Bland Altman method). Quantitative CT predicted postoperative FEV1 (r = 0.97, P < 0.001) and VC (r = 0.93, P < 0.001) well in all patients, however. CONCLUSIONS: Quantitative CT may be an alternative tool to perfusion scan to predict postresection lung function, even in patients with borderline pulmonary function undergoing a lobectomy. PMID- 16272859 TI - Pulmonary parenchymal involvement of low-grade lymphoproliferative disorders. AB - Lymphoid tissue is a normal component of the lung. The various lymphoproliferative diseases affect the lung parenchyma. The purpose of this article is to classify various lymphoproliferative diseases and to understand their computed tomography features of pulmonary involvement. The examples include follicular bronchiolitis, lymphocytic interstitial pneumonia, plasma cell granuloma, Castleman disease, lymphomatoid granulomatosis, and mucosa-associated lymphoid tissue lymphoma. Pathologic correlation is helpful for understanding imaging findings and their pathophysiology. PMID- 16272860 TI - Hemolymphangiomatosis of the spleen: imaging features. AB - Hemolymphangiomatosis is an extremely rare entity that has never been described in the diffuse form. Its main pathologic feature is proliferation of lymphatic and vascular cells inducing interstitial and hematic lakes and then splenomegaly. We report a complete imaging study, including magnetic resonance (MR) imaging, after administration of a superparamagnetic contrast agent. Ultrasonography (US), basal MR imaging, and late computed tomography (CT) contrastographic features are quite similar to those of other diffuse benign vascular neoplasms, without real hemangioma-like enhancement during the vascular phase. Its late contrastographic pattern could be considered specific if CT and iron oxide-enhanced MR imaging are applied as complementary diagnostic tools, however. PMID- 16272861 TI - Three-phase dynamic breast magnetic resonance imaging with two-way subtraction. AB - OBJECTIVE: This study was undertaken to introduce a new breast magnetic resonance (MR) imaging technique, 3-phase dynamic MR imaging with 2-way subtraction, and to examine the morphologic and kinetic features of malignant and benign breast lesions using this technique. METHODS: In 99 breasts from 89 consecutive women with suspicious breast lesions (age range: 32-72 years, mean = 48.2 years), MR imaging was performed using a fat-saturated, sagittal, gradient-echo sequence in 3 phases (1 precontrast and 2 postcontrast scans). Two-way subtraction was performed: standard subtraction (early postcontrast minus precontrast scans) and reverse subtraction (early postcontrast minus late postcontrast scans). A radiologist interpreted all the images, described the breast lesions (using the Breast Imaging Reporting and Data System MR lexicon), and assessed the lesions prospectively as being benign or malignant. The lesions with at least 1 malignant feature were classified as malignant. The MR classifications were compared with the biopsy or follow-up results. RESULTS: Sixty-five (65.7%) breast lesions were malignant, and 34 (34.3%) were benign. Forty-two (97.7%) of the 43 cases of microinvasive or invasive ductal cancer showed malignant morphologic features, and 38 (88.4%) showed washout kinetics. Eighteen (81.8%) of the 22 cases of ductal carcinoma in situ showed malignant morphologic features, and 11 (50.0%) showed washout kinetics. Twenty-nine (85.3%) of the 34 benign lesions showed neither malignant morphologic features nor washout kinetics. The sensitivity and specificity were 92.3% and 91.2% by morphologic analysis and 93.8% and 85.3% by the combined method (morphology plus kinetic analysis), respectively. CONCLUSIONS: This study demonstrated that 3-phase dynamic breast MR imaging with 2-way subtraction and a maximum intensity projection (MIP) image is a simple and useful technique for identifying breast lesions. Although the addition of the kinetic criteria enhanced the sensitivity at the cost of specificity, radiologists can diagnose a malignant lesion showing non-mass-like enhancement more confidently using reverse subtraction imaging. Therefore, this study proposes 3-phase dynamic imaging with 2-way subtraction and an MIP image as one of the standard protocols of breast MR imaging. PMID- 16272862 TI - Multidetector computed tomography of pediatric lateral condylar fractures. AB - OBJECTIVE: The objective of this study was to compare lateral condylar fracture characterization using radiographs and multidetector computed tomography (MDCT) of the elbow and to evaluate outcomes in a group of children managed based on MDCT findings. METHODS: Unenhanced MDCT of the elbow with sagittal and coronal reformations was prospectively performed without sedation in 10 children between 1 and 16 years of age with lateral condylar fractures identified on frontal and lateral radiographs of the elbow. Two blinded readers reviewed all radiographs and MDCT images independently and in consensus. For the radiographs and MDCT images, readers graded the displacement of the lateral condyle fracture fragment and classified the fractures according to the Milch classification. Articular and epiphyseal cartilage integrity was assessed on MDCT. Patients were managed based on the consensus interpretation of the MDCT study. The frequency of interobserver agreement, discordant grading of fracture displacement and fracture classification between radiographs and MDCT images, and altered management based on the MDCT findings were determined. RESULTS: Individual readers agreed on fracture displacement in 9 patients (90%) on radiographs and in all 10 patients (100%) on MDCT. Individual readers agreed on fracture classification in 4 patients (40%) on radiographs and in 9 patients (90%) on MDCT. In 4 patients (40%), grading of fracture displacement differed between the consensus interpretation of the radiographs and MDCT images. In 6 patients (60%), fracture classification differed between the consensus interpretation of the radiographs and MDCT images. All 3 patients with fracture displacement greater than 5 mm and 1 patient with fracture displacement between 2 and 5 mm demonstrated disruption of the articular and epiphyseal cartilage on MDCT. Two patients (20%) with fracture displacement near the surgical threshold of 2 mm had altered management based on the MDCT findings. All patients progressed to complete fracture healing, with no delayed displacement or other complications. CONCLUSIONS: Multidetector computed tomography is a highly reproducible means of characterizing pediatric lateral condylar fractures and frequently demonstrates disruption of the epiphyseal and articular cartilage in patients with fracture displacement greater than 2 mm. The findings of MDCT may lead to altered treatment in patients with fracture displacement near the surgical threshold of 2 mm. PMID- 16272863 TI - Neurothekeoma in the upper extremity: magnetic resonance imaging and computed tomography findings. AB - Neurothekeomas are rare benign tumors of soft tissue that are of presumed neural sheath origin. This report describes the magnetic resonance imaging and computed tomography features of a neurothekeoma in the left forearm of a 38-year-old woman with a 2-year history of a painful mass. Magnetic resonance imaging showed a smooth-bordered ovoid lesion within the inner portion of the extensor digitorum muscle with a Hounsfield number of 15. The lesion had intermediate signal on T1 weighted images, high signal on T2-weighted images, and mild to moderate heterogeneous gadolinium contrast enhancement. PMID- 16272864 TI - Volume determination of intracranial aneurysms using 16-row multislice computed tomography angiography: a systematic in vitro analysis of different reconstruction methods. AB - OBJECTIVE: To assess multislice computed tomography (CT) angiography for volume determination of intracranial aneurysms. METHODS: Submillimetric 16-row multislice CT angiography was performed with optimized scan parameters on precision spheres and a soft carotid artery model harboring 3 aneurysms connected to a pulsatile circuit. The CT angiography images were produced using optimized techniques for axial, multiplanar reformation, maximum intensity projection, surface-shaded display, and volume-rendered images. Measurements were made with electronic precision calipers by segmentation according to the method of Cavalieri and by the use of automated volumetric analysis software. RESULTS: Segmentation resulted in precise and accurate volume estimates of aneurysms, but small volumes were underestimated and evaluation time was long (36:44 minutes). Automated volume evaluation from volume-rendered reconstructions also resulted in low measurement error, although the evaluation process was significantly faster (3:25 minutes; P < 0.0001). CONCLUSIONS: The use of an automated volume analysis tool on volume-rendered reconstructions is recommended for time-efficient volume assessment of intracranial aneurysms. PMID- 16272865 TI - Magnetic resonance imaging findings of angiosarcoma of the scalp. AB - OBJECTIVE: The purpose of this study was to investigate the magnetic resonance (MR) imaging findings of angiosarcoma of the scalp retrospectively. METHODS: Eight patients with angiosarcoma of the scalp were included in this study. All patients were examined with 1.5-T MR imaging units and commercially available head coils. RESULTS: In all 8 patients, MR images revealed thickened scalp or tumors with prolonged T1 and T2 relaxation times. They were well enhanced. T2 weighted MR imaging with fat saturation and contrast-enhanced T1-weighted MR imaging with fat saturation clearly showed tumors invading the subcutaneous fat tissue and muscles. In 4 patients, the tumors were larger on MR images than on inspection. CONCLUSIONS: Magnetic resonance imaging was useful in determining the extent of angiosarcoma of the scalp because it visualized the tumor invasion into surrounding structures that could not be seen on physical inspection. PMID- 16272866 TI - Dorsoventral extension of the talairach transformation and its automatic calculation for magnetic resonance neuroimages. AB - The Talairach transformation (TT), the most prevalent method for brain normalization and atlas-to-data warping, is conceptually simple, fast and can be automated. Two problems with the TT in the clinical setting that are addressed in this article are reduced accuracy at the orbitofrontal cortex and upper corpus callosum (CC) and unsuitability for functional neurosurgery because of incomplete scanning. To increase dorsoventral accuracy, we introduce 2 additional landmarks: the top of the CC (SM) and the most ventral point of the orbitofrontal cortex on the midsagittal slab (IM). A method for their automatic calculation is proposed and validated against 55 diversified magnetic resonance (MR) imaging cases. The SM and IM landmarks are identified accurately and robustly in an automatic way. The average error of SM localization is 0.69 mm, and 91% of all cases have an error not greater than 1 mm. The average error of IM localization is 0.98 mm, approximately three quarters of cases have an error not greater than 1 mm, and 95% of all cases have an error not larger than 2 mm. The SM is correlated (R(2) = 0.72) with the most superior cortical landmark, whereas the IM is only loosely correlated (R(2) = 0.22) with the most inferior cortical landmark. On average, the original TT overlays the atlas axial plate at -24 on the orbitofrontal cortex as opposed to the correct plate at -28. Therefore, 1-dimensional ventral scaling in the original TT is insufficient to cope with variability in the orbitofrontal cortex. The key advantages of our approach are the preserved conceptual simplicity of the TT, fully automatic identification of the new landmarks, improved accuracy of the atlas-to-data match without compromising performance, and enabled TT use in functional neurosurgery when a dorsal part of the brain is not available in the scan. PMID- 16272867 TI - Magnetic resonance imaging morphometry of the midbrain in patients with Wilson disease. AB - In this study, the focus is on midbrain magnetic resonance imaging morphometric measures (transverse diameter of the midbrain peduncle [Tp], transverse diameter of the tegmentum, anteroposterior diameter of the midbrain, interpeduncular distance [IPD], and interpeduncular angle [IPA]) in a group of 47 consecutive patients with neurologic (37 patients) and hepatic (10 patients) forms of Wilson disease (WD). Morphometric measures were significantly different between the group of patients with WD and healthy controls (51 subjects) as well as patients with Parkinson disease (15 patients) and multiple sclerosis (15 patients). Among the studied variables, IPA, Tp, and IPD were particularly useful in differentiating patients from healthy subjects (probability reaching 93%). PMID- 16272868 TI - Basal cell adenoma of the parotid gland: characteristics of 2-phase helical computed tomography and magnetic resonance imaging. AB - OBJECTIVE: The goal of this study was to report the radiologic characteristics of basal cell adenoma of the parotid gland, which is a relatively rare neoplasm. METHODS: A radiology and otolaryngology specialist reviewed the 2-phase helical computed tomography (CT) (n = 6) and/or magnetic resonance (MR) imaging (n = 2) scans of 7 patients with basal cell adenoma. The authors evaluated the imaging characteristics, including tumor size, location, contour and margin, internal density or signal intensity, contrast enhancement pattern, and presence of calcification. The imaging features were then analyzed and correlated with the pathologic findings. RESULTS: All the tumors presented as small (less than 3 cm), well-encapsulated, round or oval masses on CT or MR imaging. On the 2-phase CT scan, the mostly solid-looking tumors (n = 4) showed marked contrast enhancement on the early phase, and there was a subsequent decrease in attenuation on the delayed phase. These tumors were classified as the solid subtype on histologic examination. Meanwhile, the tumors with large cystic areas (n = 2) showed gradual and additional enhancement on the delayed phase and were classified as the tubular or trabecular subtype on pathologic evaluation. There were small spots of low attenuation in the tumors of the solid subtype, which were proved to be intratumoral microcysts in the pathologic correlation. Calcification was found in a tumor. CONCLUSIONS: Basal cell adenomas of the parotid gland present as small well-marginated tumors and appear as masses with central large cysts or solid masses with microcysts on CT and MR imaging scans. Basal cell adenomas of the parotid gland had at least 2 different enhancement patterns on the 2-phase helical CT scans, and the enhancement patterns and imaging architecture were related to the histologic subtype of the tumors. PMID- 16272869 TI - Detailed nodal features of cervical tuberculous lymphadenitis on serial neck computed tomography before and after chemotherapy: focus on the relation between clinical outcomes and computed tomography features. AB - OBJECTIVE: To investigate the relation between clinical outcomes and nodal features on computed tomography (CT) in cervical tuberculous lymphadenitis (CTBL) before and after antituberculous chemotherapy. METHODS: Fifty-six patients with CTBL underwent CT before and after a 6- or 12-month course of standard chemotherapy. Three radiologists evaluated the nodal features on serial CT retrospectively, including calcification (no/punctuate/large), necrosis (no/eccentric/central), perinodal infiltrations (no/localized/extensive), and enhancing patterns (no/peripheral/homogeneous). The clinical outcome was defined as "favorable" (n = 33) or "unfavorable" (n = 23) at the completion of chemotherapy. RESULTS: All the features on the initial CT scan did not show a statistically significant difference between the favorable and unfavorable groups. On the final CT scan, absence of necrosis (P < 0.005), no infiltration (P < 0.005), no enhancement (P < 0.008), and central enhancement (P < 0.014) were more common in the favorable group, whereas large necrosis (P < 0.005), localized and extensive infiltration (P = 0.005, P < 0.005), and peripheral enhancement were more common in the unfavorable group (P < 0.005, P = 0.009). CONCLUSIONS: Central necrosis, perinodal infiltration, and peripheral rim enhancement on the final CT scan showed differences between the 2 groups. These CT features reflecting inflammation can be useful findings for assessing treatment response. PMID- 16272870 TI - Pedunculopontine nucleus: a new target for deep brain stimulation for akinesia. PMID- 16272872 TI - Bilateral deep brain stimulation of the pedunculopontine nucleus for Parkinson's disease. AB - Gait disturbance and postural instability are some the most disabling symptoms of idiopathic Parkinson's disease and in late stage disease can be resistant to both medical and surgical therapies. We implanted bilateral deep brain stimulation electrodes into the pedunculopontine nucleus in two patients with advanced Parkinson's disease. We demonstrate for the first time that low frequency (20-25 Hz) stimulation of this nucleus significantly improves gait dysfunction and postural instability in both the 'on' and 'off' medication states. Their combined total Unified Parkinson's Disease Rating Scale score improved by 53% and motor score by 57%. No procedure or stimulation-related complications were observed. If these findings are replicated in a larger number of patients, pedunculopontine nucleus stimulation may provide the means to alleviate these disabling and otherwise treatment-resistant symptoms of advanced Parkinson's disease. PMID- 16272871 TI - Implantation of human pedunculopontine nucleus: a safe and clinically relevant target in Parkinson's disease. AB - The peduncolopontine nucleus modulates locomotor activity and dysfunction in this nucleus may be responsible for the gait and postural impairments seen in Parkinson's disease and other movement disorders. We report the first surgical exploration and implantation of deep brain stimulating electrodes of the peduncolopontine nucleus area in two Parkinson's disease patients to examine the safety and the potential benefit of chronic electrical stimulation at this site. Under local anesthesia, the peduncolopontine nucleus was approached from a coronal burr hole using a trajectory that was 78-80 degrees and 62-64 degrees on the coronal and sagittal planes. Microrecordings helped to identify neurons in peduncolopontine nucleus and the adjacent substantia nigra pars reticulata. Chronic deep brain stimulating electrodes were implanted within the peduncolopontine nucleus in a manner similar to that practiced with deep brain stimulating surgery at other targets. Peduncolopontine nucleus neurons were characterized by small and broad multiunits (230 muV, 2.5 ms, 14.6 Hz). Caudal to this area, neurons firing at higher frequency, approximately 70 Hz, characteristic of nigral neuronal discharges, were encountered, followed by 2 mm of cells similar to those recorded in the dorsal peduncolopontine nucleus area. After deep brain stimulating electrodes implantation, acute intraoperative stimulation (up to 3 V) was performed with two stimulation frequencies in each session. Stimulation at 80 Hz has little discernable effect. On the other hand, stimulation at 10 Hz fostered a subjective feeling of 'well-being' and a time locked amelioration of the clinical scores. These findings demonstrate that the stereotactic approach of peduncolopontine nucleus is safe. The target may reliably be identified by microrecordings. Low-frequency stimulation may produce acute improvements in motor function. PMID- 16272873 TI - A hind limb tourniquet induces interleukin-6 expression in a rat dorsal root ganglion. AB - We investigated the mRNA levels of interleukin-6-related genes in a rat dorsal root ganglion after application of a tourniquet to a hind limb in order to identify the molecules that are induced immediately after peripheral nerve injury at the early stage. Induction of interleukin-6 and upregulation of glycoprotein 130 mRNA expressions were observed in the ipsilateral dorsal root ganglion at 4 h after tourniquet application. Interleukin-6 protein was detected in small-sized and medium-sized dorsal root ganglion cells by immunohistochemical analysis. The induction of interleukin-6 expression is likely to play a role in the protection of injured neurons perhaps related to growth of their axons. Glycoprotein 130 might also account for the inhibitory effects following nerve injury. PMID- 16272874 TI - Meditation experience is associated with increased cortical thickness. AB - Previous research indicates that long-term meditation practice is associated with altered resting electroencephalogram patterns, suggestive of long lasting changes in brain activity. We hypothesized that meditation practice might also be associated with changes in the brain's physical structure. Magnetic resonance imaging was used to assess cortical thickness in 20 participants with extensive Insight meditation experience, which involves focused attention to internal experiences. Brain regions associated with attention, interoception and sensory processing were thicker in meditation participants than matched controls, including the prefrontal cortex and right anterior insula. Between-group differences in prefrontal cortical thickness were most pronounced in older participants, suggesting that meditation might offset age-related cortical thinning. Finally, the thickness of two regions correlated with meditation experience. These data provide the first structural evidence for experience dependent cortical plasticity associated with meditation practice. PMID- 16272876 TI - Amygdala gray matter concentration is associated with extraversion and neuroticism. AB - Using high-resolution magnetic resonance imaging and voxel-based morphometry in 41 healthy individuals, this study evaluated the association between the personality traits of extraversion and neuroticism, on the one hand, and individual differences in localized brain volume and gray matter concentration, on the other, with a special focus on the amygdala. Extraversion was positively correlated with gray matter concentration in the left amygdala, whereas neuroticism was negatively correlated with gray matter concentration in the right amygdala. Given that neuroticism is a risk factor for depression, our finding offers one explanation as to why prior structural imaging studies of depressed patients (which did not control for personality) produced conflicting findings. Furthermore, our data are consistent with the view that amygdala reduction seen in depressed patients precedes the onset of the disease, rather than being a consequence of the illness. PMID- 16272875 TI - Differential brain activation according to chronic social reward frustration. AB - Neural correlates of reward frustration are increasingly studied in humans. In line with prediction error theory, omission of an expected reward is associated with relative decreases of cerebral activation in dopaminergic brain areas. We investigated whether a history of chronic work-related reward frustration influences this reward-dependent activation pattern by means of functional magnetic resonance imaging. Solving arithmetic tasks was followed by either monetary reward or omission of reward. Hyperactivations in the medial prefrontal, anterior cingulate and dorsolateral prefrontal cortex were observed in a group of healthy adults with high susceptibility to reward frustration as compared with a group with low susceptibility. Findings indicate a compromised ability of adapting brain activation among those suffering form chronic social reward frustration. PMID- 16272877 TI - Orbitofrontal thickness, retention of fear extinction, and extraversion. AB - People differ in their personality traits and in their ability to modulate fear. Does our personality determine how well we extinguish conditioned fear responses? Or is the opposite true? Herein, we examine the relationships between personality traits, memory for fear extinction, and cortical thickness as a measure of brain structure. We found that in healthy humans, extinction retention and thickness of the medial orbitofrontal cortex are positively correlated with extraversion. Path analysis indicates that extinction retention mediates the relationship between the medial orbitofrontal cortex thickness and extraversion, thereby illustrating one path through which brain structure influences personality. PMID- 16272878 TI - Cerebellar activation patterns in deaf participants for perception of sign language and written text. AB - To address the influence of language presentation on cerebellar activation, hearing volunteers were compared with deaf individuals using functional magnetic resonance imaging during observation of German sign language and text reading. In deaf participants, German sign language revealed activation of the auditory and language-related areas including the medial temporal gyrus (BA 21) and Crus I. In normal hearing participants not understanding German sign language, Crus I was less activated during observation of German sign language but strongly activated during text reading and accompanied by activation of the medial temporal gyrus. Our results indicate that cerebellar activation in Crus I corresponds to language perception as correlated with cortical activation in language-related areas and is not dependent on the mode of language presentation. PMID- 16272879 TI - Engraftment of embryonic stem cell-derived neurons into the cochlear modiolus. AB - This study aimed to evaluate the potential of embryonic stem cell-derived neural progenitors for use as transplants for the replacement of the auditory primary neurons, spiral ganglion neurons. Mouse embryonic stem cell-derived neural progenitors were implanted into the base of the cochlear modiolus of normal or deafened guinea pigs, which contains spiral ganglion neurons and cochlear nerve fibers. Histological analysis demonstrated the survival and neural differentiation of transplants in the cochlear modiolus and active neurite outgrowth of transplants toward host peripheral or central auditory systems. Functional assessments indicated the potential of transplanted embryonic stem cell-derived neural progenitors to elicit the functional recovery of damaged cochleae. These findings support the hypothesis that transplantation of embryonic stem cell-derived neural progenitors can contribute to the functional restoration of spiral ganglion neurons. PMID- 16272880 TI - Sound-induced flash illusion as an optimal percept. AB - Recently, it has been shown that visual perception can be radically altered by signals of other modalities. For example, when a single flash is accompanied by multiple auditory beeps, it is often perceived as multiple flashes. This effect is known as the sound-induced flash illusion. In order to investigate the principles underlying this illusion, we developed an ideal observer (derived using Bayes' rule), and compared human judgements with those of the ideal observer for this task. The human observer's performance was highly consistent with that of the ideal observer in all conditions ranging from no interaction, to partial integration, to complete integration, suggesting that the rule used by the nervous system to decide when and how to combine auditory and visual signals is statistically optimal. Our findings show that the sound-induced flash illusion is an epiphenomenon of this general, statistically optimal strategy. PMID- 16272881 TI - The Grueneberg ganglion projects to the olfactory bulb. AB - The Grueneberg ganglion is a compact cluster of neurons in the rostral nasal vestibule once thought to be a component of the terminal nerve, a non-sensory nerve that does not innervate the olfactory bulb. Its strong expression of olfactory marker protein, a pan-olfactory marker, in mice led us to re-examine this conclusion. Here, we demonstrate that the Grueneberg ganglion projects axons from the nasal vestibule, along the septum, through the cribriform plate and onto the olfactory necklace domain of the olfactory bulbs where it forms glomeruli. Its expression of olfactory marker protein, combined with its direct wiring to the olfactory bulb, strongly suggest that the Grueneberg ganglion is a component of the olfactory pathway. PMID- 16272882 TI - Lack of standard N2 in elderly participants indicates inhibitory processing deficit. AB - We analyzed the cortical responses to standard stimuli presented in an auditory oddball paradigm from two groups of young and elderly participants in unattended and attended conditions. A sustained negativity (N2) followed N1 and P2 in the responses of the young participants. This negativity was reduced or absent in the elderly. With attention, the early portion of N2 (N2early) decreased in both groups, whereas the late portion (N2late) increased only in young participants. We suggest inhibitory processes at the origin of this component. The lack of N2 in the elderly participants is consistent with structural changes in the aging brain and supports the hypothesis of a specific age-related deficit in the inhibition of irrelevant information processing. PMID- 16272883 TI - Expression of cyclic nucleotide-gated channels in the rat medial vestibular nucleus. AB - The role of cyclic nucleotide-gated (CNG) channels in sensory signal transduction in retinal and olfactory cells is widely recognized, but there is increasing evidence that they also play more general functions in the central nervous system as downstream effectors of cyclic nucleotides. Here, we demonstrate the expression of the alpha-subunit of rod- and olfactory-type CNG channels (CNG1 and CNG2, respectively) in the rat medial vestibular nucleus (MVN). Nested polymerase chain reaction revealed CNG channel mRNA in the MVN, and CNG1 and CNG2 proteins were also detected by Western blotting and immunohistochemistry. Finally, electrophysiological evidence is provided suggesting that CNG channels play a functional role in the MVN. PMID- 16272884 TI - Position-specificity of facial adaptation. AB - We investigated the representation of objects' position at the higher, shape selective stages of visual processing by testing the position-specificity of the behavioural and neural effects of facial adaptation. Here, we show that facial after-effects evoked by adaptation to both upright and upside-down faces are significantly larger when the adaptor and test faces are presented on the same retinal position than when they are displayed in different hemifields. Our event related potential recordings revealed that adaptation effects measured on the amplitude of the N170 event-related potential component over the hemisphere that was contralateral to the test face stimulus also show strong position specificity. These findings suggest that face adaptation effects are only partially translation invariant and facial after-effects measured with peripheral test stimuli primarily reflect the adaptation processes in the contralateral hemisphere. PMID- 16272885 TI - The voltage dependence of GABAA receptor gating depends on extracellular pH. AB - Recent studies have indicated that changes in extracellular pH and in membrane voltage affect the gamma-amino-n-butyric acid type A receptor gating mainly by altering desensitization and binding. To test whether the effects of membrane potential and pH are additive, their combined actions were investigated. By analyzing the current responses to rapid gamma-amino-n-butyric acid applications, we found that the current to voltage relationship was close to linear at acid pH but the increasing pH induced an inward rectification. Desensitization was enhanced at depolarizing potentials, but this strongly depended on pH, being weak at acidic and strong at basic pH values. A similar trend was observed for the onset rate of responses to saturating gamma-amino-n-butyric acid concentration. These data provide evidence that the voltage sensitivity of GABAA receptors depends on extracellular pH. PMID- 16272886 TI - Influence of the point mutation alpha-1-H101R on the assembly of gamma aminobutyric acid type A receptors. AB - The point mutation H101R in the alpha1-subunit of gamma-aminobutyric acid type A receptors is known to abolish effects by benzodiazepine diazepam. This mutation and homologous mutations in other alpha-subunits have been used to quantify receptor pentamers containing two different alpha-subunit isoforms, and to study the role of alpha-subunit isoforms in the response of mice to diazepam. Both types of study assumed implicitly or explicitly that this mutation strongly affects assembly with the gamma2-subunit. Here, we investigated the assembly properties of mutated in comparison with wild-type subunits, and demonstrate that alpha1H101R has similar assembly properties as wild-type alpha1. PMID- 16272887 TI - Analysis of hedgehog interacting protein in the brain and its expression in nitric oxide synthase-positive cells. AB - Hedgehog interacting protein (Hip) and Patched 1 (Ptc1) regulate the cell responses to the morphogen Sonic Hedgehog (Shh). Here, we compare the relative expression patterns of Shh, Hip and Ptc1 transcripts in the E13.5 mouse brain embryo. We observe that the expression of Hip and Ptc1 often overlaps and is found close to Shh-expressing cells, suggesting that both proteins are required for controlling Shh signals. In the adult striatum in which Ptc1 is not detected, we show that a majority of Hip-expressing cells correspond to neurons expressing the neuronal form of nitric oxide synthase. These data raise the hypothesis for a functional link between nitric oxide and Shh signaling and for a nonredundant role of Hip and Ptc1 in the adult brain. PMID- 16272888 TI - Neuroprotective effects of candesartan against cerebral ischemia in spontaneously hypertensive rats. AB - The cerebral protective effects of 4-week treatment with candesartan (0.3, 1, 3 mg/kg/day) and ramipril (0.5, 1.5, 5 mg/kg/day) were examined in spontaneous hypertensive rats 24 h after middle cerebral artery occlusion. We found that both candesartan and ramipril could reduce the infarct volume and neurological deficit scores compared with control. Importantly, the neuroprotective effects of candesartan (1 mg/kg/day) were abolished by PD123319 (an AT2 receptor antagonist, 10 mg/kg/day). AT1 receptor gene expression was downregulated while AT2 receptor gene expression was upregulated by candesartan. It is concluded that candesartan appears to provide beneficial effects against stroke in spontaneous hypertensive rats in three ways: AT1 receptor antagonism, downregulation of AT1 receptor expression and upregulation of AT2 receptor expression. PMID- 16272889 TI - PP2 inhibits glutamate release from nerve endings by affecting vesicle mobilization. AB - Src kinase is widely expressed in the brain and its inhibition with PP2 has previously been shown to depress depolarization-evoked glutamate release from rat cerebrocortical synaptosomes by reducing voltage-dependent Ca2+ entry. In this study, we further showed that the inhibitory effect of PP2 on 4-aminopyridine evoked glutamate release results from a reduction of vesicular exocytosis and not from an inhibition of non-vesicular release. In addition, PP2 significantly inhibited ionomycin-induced or hypertonic sucrose-induced glutamate release. Also, disruption of cytoskeleton organization with cytochalasin D occluded the inhibitory action of PP2 on 4-aminopyridine and ionomycin-evoked glutamate release. These results suggest that PP2-mediated inhibition of glutamate release involves the modulation of some exocytotic steps, possibly through a regulation of actin cytoskeleton dynamics. PMID- 16272890 TI - The effect of electromagnetic fields emitted by mobile phones on human sleep. AB - Previous research has suggested that exposure to radiofrequency electromagnetic fields increases electroencephalogram spectral power in non-rapid eye movement sleep. Other sleep parameters have also been affected following exposure. We examined whether aspects of sleep architecture show sensitivity to electromagnetic fields emitted by digital mobile phone handsets. Fifty participants were exposed to electromagnetic fields for 30 min prior to sleep. Results showed a decrease in rapid eye movement sleep latency and increased electroencephalogram spectral power in the 11.5-12.25 Hz frequency range during the initial part of sleep following exposure. These results are evidence that mobile phone exposure prior to sleep may promote rapid eye movement sleep and modify the sleep electroencephalogram in the first non-rapid eye movement sleep period. PMID- 16272891 TI - Eliminate guessing games with consistency. PMID- 16272892 TI - Amp up your leadership curiosity. AB - Employ curiosity as a way to relinquish control and empower employees. PMID- 16272893 TI - Get organized with an orientation database. AB - Information generated from a database can be used to monitor and improve an orientation program. PMID- 16272894 TI - For leadership effectiveness, look inside. AB - Learn to develop a personal model of leadership effectiveness. PMID- 16272895 TI - Measure patient harm in real time. AB - A California healthcare system incorporates clinical information technology to mine data for adverse events. PMID- 16272897 TI - Triple threat: diabetes, hypertension, and heart disease. AB - Review care specifics for prediabetes, including details of hypertension and metabolic syndrome. PMID- 16272896 TI - Best-practice protocols: Preventing surgical site infection. AB - Inconsistent application of infection control practices may contribute to differences in SSI rates and mortality of surgical patients. PMID- 16272899 TI - Transfer admission discharge teams keep things moving. AB - After implementing a transfer, admission, and discharge team, one facility experiences improvements in patient care, productivity, and ED diversions, and heightened patient, nurse, and physician satisfaction. PMID- 16272900 TI - Better project management: better patient outcomes. AB - Review production concepts specific to nurse managers. PMID- 16272901 TI - Ensure patients' wounds are best dressed. AB - Get the latest information on products specifically designed for effective wound management. PMID- 16272904 TI - Tips for top challenges: find strength in emotional leadership. PMID- 16272905 TI - Recalling our history as we look toward the future ... PMID- 16272907 TI - Time for a humanity check: nurses connecting ... nurses making a difference. PMID- 16272908 TI - The newborn foot: diagnosis and management of common conditions. AB - An assessment of the foot should be a part of every newborn physical examination. The newborn foot, although complex in structure, can be examined quickly in any office setting. Many foot deformities are diagnosed immediately after birth, allowing for earlier and often more successful treatment. A neonate with a foot deformity can be a source of anxiety to parents. Adequate knowledge of commonly encountered neonatal foot problems enables the nurse to give appropriate anticipatory guidance to the parents. Foot abnormalities usually occur as isolated findings in an otherwise healthy newborn. However, these deformities can also be seen in newborns with underlying neuromuscular disorders and syndromic conditions. Common newborn foot abnormalities include equinovarus deformity (clubfoot), metatarsus adductus, calcaneovalgus, congenital vertical talus, polydactyly (supernumerary digits), and syndactyly (webbed toes). These common foot disorders, their treatment, and their prognosis are discussed. This article also discusses fundamental nursing implications needed to care for these young patients and their families. PMID- 16272910 TI - Multiple organ dysfunction syndrome after multiple trauma. AB - Multiple organ dysfunction syndrome (MODS) is a serious, often fatal, sequel to multiple trauma. Prolonged shock states, severe sepsis, acute pancreatitis, acute renal failure, severe burns, ischemic conditions, and acute respiratory distress syndrome have been implicated in the development of MODS. Despite advances in critical care therapies and infection control practices, severe sepsis remains a major health problem, with an estimated mortality rate of 30-50% (Ely, Kleinpell, & Goyett, 2003). The purposes of this article are to describe the pathophysiologic changes that lead to the development of MODS and discuss strategies to prevent the development of MODS and to treat MODS if it develops. PMID- 16272912 TI - Health-related stressors experienced by patients who underwent total knee replacement seven days after being discharged home. AB - BACKGROUND: Empirical evidence indicates that the first few weeks after total knee replacement are likely to be stressful for patients. However, no reported studies have examined the type of stressors that individuals encounter during the first few days at home. PURPOSE: The purpose of this study was to describe stressors experienced by patients 7 days post-hospitalization after total knee replacement who were managing their care at home. METHOD: The stressor of most concern to the patient was identified using an open-ended question. Intensity, frequency, and duration of the stressors were measured using five-point Likert response scales. RESULTS: Various stressors were identified, but those of most concern related to pain management and bowel difficulties. Respondents indicated the stressors caused a great deal of concern/worry and occurred daily and most could have been prevented if patients had sufficient self-care knowledge. IMPLICATIONS: The study suggests that preoperative education of patients may need to be improved and that nurses may need to contact patients after discharge to assist them with postoperative problems and concerns. PMID- 16272913 TI - The evolution of orthopaedic nursing at the Hospital for Special Surgery: the first orthopaedic institution in the United States. AB - The history of nursing began in London in the late 1800s with the reform of unsanitary conditions by Florence Nightingale. During the same period, the United States was bitterly fighting the Civil War. Nursing had not developed as a profession, and most of the duties performed by nurses were conducted by men. Casualties of war required rehabilitation and care. Crippled children were left to die because they were considered a burden to society. Dr. James Knight founded the Hospital for the Ruptured and Crippled in his home on Second Avenue. This would later become a world-renowned orthopaedic institution with exceptional nursing care. A historical analysis of nursing education and practice are reviewed, along with the evolution of the first orthopaedic hospital in the United States. PMID- 16272914 TI - Multicenter pin care study. AB - BACKGROUND: Pin-site infection is a common complication of external fixation. Because few studies have compared methods of pin care that reduce infection rate, there is a need for evidence-based practice guidelines for pin-site care. METHODS: Two of 10 original clinical centers completed a prospective, randomized pin-care study between May 2000 and May 2002 to determine which of seven methods for caring for skeletal pins (external fixator, traction, or halo) resulted in the fewest pin-site infections. RESULTS: The 92 subjects had an average infection rate of 34%, and the 527 pins had a rate of 20%. Thirty patients (98 pins) had stage II infections, two patients (12 pins) had stage III infections, and none had deep infection or osteomyelitis. The protocols were (1) half-strength peroxide cleansing and gauze wraps (45%), (2) half-strength peroxide cleansing and Xeroform wraps (9%), (3) saline cleansing and gauze wraps (33%), (4) saline cleansing and Xeroform wraps (26%), (5) antibacterial soap-and-water cleansing and gauze (38%), (6) antibacterial soap-and-water cleansing and Xeroform gauze (50%), and (7) stable dressings with no pin cleansing (36%). Logistic regression analysis demonstrated significant inverse relationships (p = .05) between infection rate and age, as well as fixator type; the latter may be related to exposed threads. CONCLUSION: Results suggest that other factors outside the realm of this study may affect children's pin-site infection rate and that half strength peroxide and Xeroform dressings were superior to soap-and-water cleansing. This pilot study indicates a need for further research with a larger sample size and for exploring factors in a younger population. PMID- 16272915 TI - Treatment options for articular cartilage defects of the knee. AB - The treatment of symptomatic articular cartilage defects of the knee has evolved tremendously in the past decade. Previously, there were limited treatment options available to patients who suffered from either partial-thickness or full thickness cartilage lesions. Because articular cartilage has a limited capacity for healing, patients were often treated symptomatically until they became candidates for osteotomy or total joint replacement. Recently, both reparative and restorative procedures have been developed to address this significant source of morbidity in young active patients. Microfracture is a reparative technique that induces a healing response to occur in an area of articular cartilage damage. Osteochondral autografts and allografts in addition to autologous chondrocyte implantation are restorative techniques aimed at recreating a more normal articular surface. Both types of procedures have been developed to alleviate the symptoms associated with focal chondral defects, as well as limit their potential to progress to a diffuse degenerative arthritis. Treatment can vary depending on both cartilage defect and patient factors. This article summarizes the various treatment options that have recently become available. PMID- 16272918 TI - Do your patients understand? Determining your patients' health literacy skills. AB - Despite teaching endeavors, nurses are constantly faced with patients who do not understand how to manage their healthcare. This problem has come to the forefront of healthcare issues. As a society, there is concern that despite medical advances, progress with healthcare may be in jeopardy because the skills needed by patients to manage their care are insufficient. This issue is affected by many factors. One of the most prominent factors is the lack of patient health literacy skill assessment. One of the first and most basic parts of the nursing process is to assess the patient. To teach patients, we must identify their learning needs, but the assessment cannot stop there. Nurses need to know patients' health literacy skills so that they can teach them in the best manner possible. This article provides specific information on health literacy assessment tools and the skills needed by nurses to use these tools. Each nurse must decide what tools will work for his or her patients, so that in the end, each patient will understand how to manage his or her healthcare. PMID- 16272920 TI - Fighting infection: an ongoing challenge, part 3-antimycobacterials, antifungals, and antivirals. AB - The first part of this series provided a brief overview of how antimicrobials, the "silver bullets" of modern medicine, are designed to target specific agents of infection. The second part addressed several classes of antibacterials: bacterial cell wall inhibitors (penicillins, cephalosporins), protein synthesis inhibitors (macrolides, tetracyclines, aminoglycosides), and nucleic acid inhibitors (sulfonamides and quinolones). This third section focus on those drugs used to treat mycobacterial infections (Mycobacterium tuberculosis), fungal infections (Candida species, Aspergillus species), and viral infections (herpes, hepatitis, and influenza). Selected agents in each classification are identified, with a look at the specific use, action, and potential for adverse effects. A brief comment about the challenges for future development of antimicrobials is included. PMID- 16272922 TI - European consensus statement on cochlear implant failures and explantations. PMID- 16272923 TI - Fresh tympanic membrane perforations heal without significant loss of strength. AB - HYPOTHESIS: The mechanical and structural properties of the tympanic membrane change after a perforation has healed. BACKGROUND: In previous studies, efforts have been made to enhance the healing process of tympanic membrane perforations. The strength of the healed perforation has been tested with moire interferometry in gerbils, but in no other species. METHODS: A laser myringotomy was made on 10 Sprague-Dawley rats and 10 CBA mice, and assessments were made after 2 or 4 weeks with moire interferometry and light and electron microscopy. RESULTS: The mean peak displacement at pressure loads of +350 daPa and of -350 daPa did not differ significantly in the healed perforations as compared with the untouched tympanic membranes. Morphologic assays showed fivefold increased thickness at the site of the perforation due to invaded fibroblasts and extracellular matrix. CONCLUSION: Moire interferometry was successfully performed in the rat ears, whereas in mouse ears the method was not easily applicable due to technical difficulties. The stress-strain curve of the rat tympanic membrane displays an S-shape. The strength of the spontaneously healed tympanic membrane after myringotomy was not significantly impaired. The site of the perforation became significantly thickened at 2 and 4 weeks post-myringotomy. This information is of clinical importance, because recently closed perforations will be challenged by pressure gradient in everyday life. PMID- 16272924 TI - Mastoid and epitympanic obliteration in canal wall up mastoidectomy for prevention of retraction pocket. AB - OBJECTIVE: To evaluate the surgical outcome in patients who have undergone mastoid and epitympanic obliteration technique. STUDY DESIGN: : Retrospective review. SETTING: Tertiary care referral center. PATIENTS: The study group included 151 patients with cholesteatoma who underwent mastoid and epitympanic obliteration technique. This technique was applied to cases who had a strong chance of recurrent retraction pocket and cholesteatoma formation, including those patients with an adhesive drum indicating poor eustachian tube function (n = 52) and patients with a destructive scutum (n = 68). Thirty-one patients had both an adhesive drum and a destructive scutum. INTERVENTION: The connection between the mastoid cavity and the middle ear was blocked by obliterating the epitympanum and antrum with bone pate and the remaining mastoid cavity with abdominal fat. MAIN OUTCOME MEASURES: The postoperative drum state, the incidence of retraction pocket formation and cholesteatoma recurrence, the surgical complications of obliteration, and the hearing outcome. RESULTS: In 114 of 151 patients (75.4%), the middle ear was well healed and well aerated. The retraction pocket formation or cholesteatoma recurrence did not develop in any subject. Postauricular skin depression was the most common complication of this technique (n = 31 [20.5%]). In three patients (2.0%), the bone pate used for obliteration was infected. Of the 56 cases who underwent a staged operation to regain their hearing, 37 resulted in a postoperative air-bone gap less than 20 dB hearing level. CONCLUSION: Mastoid and epitympanic obliteration is an effective option for preventing a retraction pocket and cholesteatoma recurrence in patients with a poorly functioning eustachian tube or a defective scutum, while preserving the same advantage of the canal wall up technique. PMID- 16272925 TI - Endoscopic examination of the eustachian tube: a step-by-step approach. AB - OBJECTIVE: The aim of this study was to develop a step-by-step approach for endoscopic examinations of the eustachian tube on awake patients and to report anatomic and functional findings. STUDY DESIGN: Prospective study. SETTING: University hospital. PATIENTS: Convenience sample of seven individuals without a history of ear disease. INTERVENTION: Diagnostic transnasal-transpharyngeal videoendoscopy of the eustachian tube with 30- and 70-degree rigid Hopkins rod endoscopes, 2.5- and 0.8-mm, 0-degree flexible fiber endoscopes performed under local anesthesia in 12 eustachian tubes. MAIN OUTCOME MEASURES: Utility of the various endoscopes for the diagnosis in the different parts of the eustachian tube; quality of vision and the patient's comfort during the procedure. RESULTS: The 2.5-mm flexible endoscope was most useful for examination of the pharyngeal ostium and the cartilaginous lumen of the tube. The isthmus region could only be passed using an 0.8-mm fiberscope. In all cases, it was possible to insert the endoscope into the middle ear cavity. Eleven of the 12 tube examinations showed normal findings. The mobility of the tubal cartilage could be visualized with sufficient quality. In 50% of all examinations, application of local anesthesia via a tube catheter was necessary to make the procedure tolerable. CONCLUSION: The presented approach allows an assessment of both anatomic and functional changes to the eustachian tube in awake patients. The assessment of middle ear structures is limited. To ensure a comfortable and safe procedure, the use of topical anesthesia in a supine position and, in certain cases, additional anesthesia via eustachian tube catheter is recommended. PMID- 16272926 TI - Ultrastructural analysis of the chorda tympani nerve in chronic suppurative otitis media. AB - OBJECTIVE: To compare ultrastructure of the chorda tympani nerve by light and electron microscopy in patients with otosclerosis and chronic suppurative otitis media. STUDY DESIGN: Comparative prospective study. SETTING: Tertiary care, referral medical center. PATIENTS: The tympanic segments of chorda tympani nerves were collected for ultrastructural investigation in 20 cases with chronic suppurative otitis media and 10 cases with otosclerosis that underwent middle ear surgery. RESULTS: Histopathologic examinations of the suppurative group showed that unmyelinated fibers were almost totally lost and replaced by collagen fibers. The thickness of the myelin sheaths was very slender, whereas some of them were atrophic. A substantial increase was encountered in the endoneural collagen substance and connective tissue, whereas inflammatory elements and edema were present occasionally. Degenerative alterations of the myelinated fibers mainly occurred in the form of adaxonal vacuoles. There was disorganization and separation of parallel lamellae of Schmidt-Lanterman clefts. CONCLUSION: The chorda tympani nerve should be preserved in otologic surgery. However, inadvertent dissection of the chorda tympani nerve in chronic suppurative otitis media surgery will not possibly cause a postoperative disturbance in light of ultrastructural changes that occur in the nerve. PMID- 16272927 TI - The ponticulus: an anatomic study. AB - HYPOTHESIS: The purpose of this study was to resolve anatomic ambiguities of the ponticulus so the surgeon will be better able to remove disease from the posterior tympanum. BACKGROUND: The first step in refining an operative approach is to acquire a thorough understanding of the anatomy. A detailed study of one structure of the posterior tympanum, the ponticulus, has not yet been reported. METHODS: Fifty temporal bone plugs harvested from human cadavers were studied. The presence or absence of the ponticulus, its configuration, and its proximity to other middle ear structures were recorded for each temporal bone. RESULTS: There was complete formation of the ponticulus in 33 of the 50 specimens. In 7, only a remnant of the ponticulus formed, and it was completely absent in 10 of the 50 specimens. In the 33 specimens in which the ponticulus was present, it measured less than 1 mm in thickness in 22, it was larger than 1 mm in 7, and in 4 specimens it extended down to the floor of the middle ear as a sheet of bone. In 8 specimens, the ponticulus lay just adjacent to the stapes. CONCLUSION: The ponticulus is present in the majority of ears. It is best defined as a bridge of bone from the pyramidal eminence to the promontory and most commonly occurs as a thin, bony structure. This definitive study of the ponticulus will allow the otologic surgeon to better remove disease from the posterior tympanum. PMID- 16272928 TI - Possible role of Helicobacter pylori in the etiopathogenesis of chronic otitis media. AB - OBJECTIVE: To investigate a possible contribution from Helicobacter pylori to the etiopathogenesis of chronic otitis media. STUDY DESIGN: Prospective clinical trial. METHODS: Tympanomastoidectomy, radical mastoidectomy, and modified radical mastoidectomy procedures were performed according to the pathology of ear disease. Biopsy specimens were taken from middle ear, mastoid antrum, and tympanic orifice of eustachian tube. Helicobacter pylori DNA was extracted from these biopsy specimens by using nested polymerase chain reaction. RESULTS: Helicobacter pylori DNA was detected in 3 (7.9%) of 38 chronic otitis media patients. CONCLUSION: Even though it is possible to detect Helicobacter pylori in middle ear cleft in chronic otitis media, its role in the etiopathogenesis of the issue is controversial. PMID- 16272929 TI - Histologic otosclerosis is associated with the presence of measles virus in the stapes footplate. AB - HYPOTHESIS: Persistent measles virus infection of the otic capsule is presumed to be one of the etiologic factors in otosclerosis. The viral pathogenesis of otosclerosis could be established only by correlative analysis: histologic examination of the stapes footplates and reverse-transcriptase polymerase chain reaction amplification of the viral RNA. At present, histologic analysis of the removed stapes footplates is the only appropriate method of distinguishing otosclerotic and nonotosclerotic stapes fixations. BACKGROUND: The presence of measles virus was shown in otosclerotic patients by reverse-transcriptase polymerase chain reaction amplification of the viral RNA and detecting the viral proteins by immunohistochemistry. METHODS: Nucleic acids (mRNA, vRNA, and DNA) were extracted from ankylotic stapes footplates of stapes fixation patients (n = 44). Measles virus genomic nucleoprotein RNA was amplified by seminested reverse transcriptase polymerase chain reaction. Amplification results were correlated to postoperative histologic findings. RESULTS: Measles virus RNA was detectable only in histologically otosclerotic stapes footplates (n = 32). Histology for virus negative footplates (n = 12) excluded otosclerosis. Virus-negative stapes footplates showed annular calcification (n = 8), bone resorption with increased numbers of hemosiderophages (n = 2), and mononuclear cell infiltration with osteolysis (n = 2). CONCLUSION: Stapes ankylosis is a heterogenous disease causing conductive hearing loss with different causes. Nonotosclerotic stapes fixations may belong to degenerative disorders with variable histopathology. Otosclerosis is an inflammatory disease resulting from persisting measles virus infection of the otic capsule. PMID- 16272930 TI - Video-oculography findings in patients with otosclerosis. AB - OBJECTIVE: To evaluate the existence of vestibular irritation with video oculography before and after stapes surgery and to examine whether there would be signs of specific end-organ irritation. STUDY DESIGN: A prospective study of preoperative and postoperative nystagmus, vertigo, and hearing thresholds. SETTING: University hospital, tertiary referral center. PATIENTS: Thirty-three patients (mean age, 47 yr) with otosclerosis. INTERVENTION: Stapedotomy/stapedectomy with laser or microdrill. MAIN OUTCOME MEASURES: Spontaneous, gaze-evoked, and head-shaking nystagmus was measured preoperatively and approximately 1 week, 1 month, and 3 months after the operation. Three dimensions of nystagmus were identified and their slow-phase velocities were calculated. RESULTS: Spontaneous horizontal nystagmus was found preoperatively in 18% (slow-phase velocities, 1.3-3.3 deg/s) and postoperatively in 11 to 19% of the patients (slow-phase velocities, 1.3-3.8 deg/s). Head-shaking nystagmus was not detected preoperatively. After the operation, 11 to 15% of the patients had head-shaking nystagmus (slow-phase velocities, 6.6-17.8 deg/s), but this prevalence did not differ statistically significantly from the preoperative level (p = 0.18). Vertical nystagmus was found equally pre- and postoperatively. Torsional nystagmus was not found. One week after the operation, nine patients (27%) had some sensation of vertigo, but it lasted over 1 month in only one patient. We found no significant correlation with vertigo and the types of nystagmus. CONCLUSION: Nystagmus with a low slow-phase velocity can occur in patients with otosclerosis. However, according to the video-oculographic findings and subjective symptoms, significant vestibular dysfunction seems to be rare and temporary after stapes surgery. PMID- 16272931 TI - Bone-conduction hearing and the occlusion effect in otosclerosis and normal controls. AB - OBJECTIVE: The goal of this study was to better understand bone-conduction hearing in subjects with normal hearing and in those with otosclerosis through the occlusion effect. With this study, the authors hope to lend credence to commonly accepted theories of bone-conduction hearing and the effect of lateralization during the Weber tuning fork test. BACKGROUND: There are three accepted theories defining bone-conduction hearing: compressional bone conduction describes an auditory percept produced by the compression and expansion of the cochlea leading to basilar membrane vibration; inertial bone conduction describes the inertia of the ossicular chain as a result of skull vibration during bone conduction testing; whereas skull vibration may also be transmitted to the external auditory canal, surrounding soft tissues, and para-auditory structures to illicit tympanic membrane vibration known as osseotympanic bone conduction. METHODS: Twenty normal volunteers and 17 unilateral otosclerosis patients underwent external canal sound pressure level measurement during bone-conduction testing using a standardized bone oscillator placement and stimulation paradigm. Sound was detected with a probe microphone placed in the external auditory canal in nonoccluded and occluded conditions after a 50-dB hearing level bone conduction stimulus. RESULTS: There was no significant difference in sound pressure level between otosclerosis and normal subjects when the external auditory canals were nonoccluded. With occlusion, sound pressure level increased in both groups, but at a statistically significantly higher level for the otosclerosis group. CONCLUSION: Sound measured in the external canal likely represents energy lost to the environment transmitted through the middle and external ear systems, aided by the effect of both inertial and osseotympanic bone conduction. Occluding the ear leads to sound trapping and amplification. Also, the pressure exerted against the tympanic membrane reduces middle ear compliance and increases the impedance mismatch between air and the middle ear system, reflecting sound back into the external canal. This effect is further enhanced by stapes fixation to explain our data in both groups of subjects. The final common pathway in "lateralization" is probably a product of higher than normal impedance mismatch at the oval window. PMID- 16272932 TI - Success rate in revision stapes surgery for otosclerosis. AB - OBJECTIVE: The aim of this study was to evaluate the hearing results of revision stapes surgery performed because of previously failed operations and to determine the causes of failure. STUDY DESIGN: Retrospective review of revision stapes operations. SETTING: Tertiary referral center. PATIENTS: Sixty-three consecutive revision stapes operations were performed in 56 patients over a period of 12 years (1992-2004). The indication for revision surgery was recurrent or persistent air-bone gap greater than 20 dB after primary surgical treatment of otosclerosis of the oval window. RESULTS: All patients were operated on to improve hearing. Sixty-three revision stapes operations resulted in closure of the air-bone gap to 10 dB or less in 52.4% of cases. The average postoperative air-bone gap was 13.1 dB, and the mean pure-tone average improvement was 12.9 dB. In six patients (9.5%), revision surgery produced no change in hearing, and in four (6.3%) the hearing decreased by 5 dB or more. In one patient, the operation resulted in a profound hearing loss. Prosthesis malfunction was the most common primary cause of failure (60.3%). The original prosthesis was replaced with a new one in 48 cases. In 30 of these (62.5%), closure of the air-bone gap to within 10 dB was achieved. In 15 cases, the prosthesis was not replaced, and in only four of these (26.7%), closure of the air-bone gap within 10 dB was obtained (p < 0.022). CONCLUSION: Revision stapes surgery is less likely to be successful than the primary operation. Closure of the air-bone gap to within 10 dB was achieved in 52.4% of patients. The success rate was better in cases where the original prosthesis was replaced with a new one. The risk for decreased bone-conduction threshold does not seem to be higher than in primary surgery. PMID- 16272933 TI - Pulsatile tinnitus: a review of the literature and an unusual case of iatrogenic pneumocephalus causing pulsatile tinnitus. AB - BACKGROUND: Pulsatile tinnitus is frequently attributed to identifiable and treatable causes, in contrast to the more common subjective non-pulsatile tinnitus. It usually originates from vascular structures as a result of either increased blood flow or lumen stenosis; atherosclerotic carotid or subclavian artery disease; arterial, venous, or arteriovenous malformations, fistulas, or dissection; and paragangliomas. Other causes have also been reported, with often unclear pathophysiology. OBJECTIVE: The aim of this paper is to present a case of pulsatile tinnitus secondary to iatrogenic pneumocephalus and to review the literature on pulsatile tinnitus. SUBJECT: A 48-year-old white woman had a roaring, very disturbing, pulsatile tinnitus after the removal of a cerebellar lobe meningioma. When the patient experienced the symptom of tinnitus, a pulsatile movement of the tympanic membrane could be clearly seen, and this was synchronous with the patient's heartbeat. Computed tomography revealed an epidural pneumocephalus in the left posterior fossa communicating freely with the air cell system of the left mastoid cavity without any sign of residual tumor. A simple mastoidectomy was performed. The whole air cell system was removed and the mastoid cavity was filled with abdominal fat. After the operation, the pulsatile tinnitus ceased completely and the pneumocephalus disappeared gradually. The patient is free of symptoms 11 months after surgery. CONCLUSION: Otologists, neurosurgeons, and skull base surgeons should be aware of this surgical complication and be careful to identify any accidental opening to the air cell system of the temporal bone and meticulously close it when it happens. The review of the literature leads to the conclusion that pulsatile tinnitus should be thoroughly investigated, as it may be related to diseases that may have serious complications. PMID- 16272934 TI - Long-term functional outcomes and academic-occupational status in implanted children after 10 to 14 years of cochlear implant use. AB - OBJECTIVES: To assess a group of consecutively implanted children over 10 years after implantation with regard to implant device use and function, speech perception, and speech intelligibility outcomes; and to document current academic or occupational status. STUDY DESIGN: A prospective longitudinal study assessing device function, device use, speech perception, speech intelligibility, and academic/occupational status of implanted deaf children. SETTING: Pediatric tertiary referral center for cochlear implantation. METHODS: The auditory performance and speech intelligibility development of 30 profoundly deaf children were rated before cochlear implantation and at 5 and 10 years after implantation using the Categories of Auditory Performance and the Speech Intelligibility Rating. The academic and/or occupational status of the participants after 10 years of implant experience was documented. All children received a Nucleus multichannel cochlear implant between the ages of 2.5 and 11 years (mean age at implantation, 5.2 yr). Implant experience ranged from 10 to 14 years of use. RESULTS: After 10 years of implant experience, 26 subjects (87%) reported that they always wore their device; 2 subjects (7%), frequently; and 1 subject (3%), occasionally. Only one child had discontinued use of his device. After 10 years of implant use, 26 (87%) of the children understood a conversation without lip reading and 18 (60%) used the telephone with a familiar speaker. Ten years after implantation, 23 (77%) of the subjects used speech intelligible to an average listener or a listener with little experience of a deaf person's speech. One third to one-half of the implanted children continued to demonstrate improvements at 5 to 10 years of implant use. Of the 30 implanted children, 8 (26.7%) experienced nine device failures. The length of time from identification of the first faulty electrode to reimplant surgery ranged from 2 weeks to 5.5 years, as several failures were gradual or intermittent. However, all children were successfully reimplanted. At the end of the study (10-14 yr after implantation), 19 subjects were in secondary school for children aged 11 to 16 years: 6 were in mainstream schools, 7 were in specialist hearing-impaired units attached to a mainstream secondary school, and 6 were in schools for the deaf. Of the remaining 11 subjects, 4 were in college studying vocational subjects, 2 were in a university studying for a bachelor's degree, 3 were working full-time, 1 was working and going to a university part-time, and 1 was a full-time mother of two young children. CONCLUSION: All but 1 of the 30 implanted children continue using their devices 10 to 14 years after implantation, showing significant progress in speech perception and production. Device failure was frequent, but successful reimplantation occurred in all cases. One-third to one-half of the implanted children in this study continued to demonstrate improvements at 5 to 10 years of implant use. All children are studying or working and are actively involved in their local communities. The results suggest that cochlear implantation provides long-term communication benefit to profoundly deaf children that does not plateau for some subjects even after reimplantation. This study further indicates that cochlear implant centers need the structure and funding to provide long-term support, counseling, audiologic follow-up, rehabilitation, and device monitoring to implanted children. PMID- 16272935 TI - Evaluation of the advance off-stylet insertion technique and the cochlear insertion tool in temporal bones. AB - OBJECTIVE: The concept and design of new cochlear implant electrodes is a challenging process. To evaluate new electrode designs, we present a study that uses a microgrinding procedure to evaluate damage to the microstructures of the cochlea resulting from the insertion procedure. In this study, we compared different insertion techniques with the Contour electrode with Softip for placement inside the cochlea and any resulting damage. METHODS: Twenty-five fresh frozen human temporal bones were used to compare electrode insertion characteristics with three insertion techniques (i.e., conventional insertion, Advance Off-Stylet performed manually, and Advance Off-Stylet performed with insertion tool) and two prototype variants of the Contour electrode with Softip (referred as Softip I and Softip II in this article). Five temporal bones were used for each arm of the study: Softip I electrode and conventional insertion; Softip I electrode and manual Advance Off-Stylet insertion; Softip I electrode and Advance Off-Stylet insertion with an early experimental insertion tool; Softip II prototype electrode and manual Advance Off-Stylet insertion; and Softip II prototype and Advance Off-Stylet insertion with a prototype insertion tool. The temporal bones were dehydrated and embedded in epoxy and used for the microgrinding procedure. Resulting images were documented and compared with conventional radiographic images. RESULTS: Our results showed that, especially when using the conventional insertion technique with Softip I electrode arrays, basilar membrane perforations were observable. Using the prototype insertion tool, good placement of the electrode array but also two basilar membrane perforations (one with each type of electrode) were observed. In contrast, the Advance Off-Stylet insertion technique did not show basilar membrane perforation with Softip I and II electrodes and resulted in reliable perimodiolar placement of the arrays. CONCLUSION: Using microgrinding of temporal bones, the Advance Off Stylet insertion technique was proven to enable more atraumatic insertions of Contour electrodes with Softip and to provide very reliable perimodiolar placements. PMID- 16272936 TI - Diagnosis and management of spontaneous cerebrospinal fluid otorrhea. AB - OBJECTIVE: To describe the clinical presentation, diagnosis, surgical management, and outcome of patients with spontaneous cerebrospinal fluid otorrhea. STUDY DESIGN: Retrospective case series. SETTING: Tertiary referral center. METHODS: The authors conducted a chart review of all previously unreported cases of surgically confirmed cerebrospinal fluid otorrhea at their institution between September 1996 and February 2005. Acquired cases were excluded from this study. Eleven cases of spontaneous cerebrospinal fluid otorrhea were identified among 10 patients. RESULTS: Nine of the 10 patients presenting with spontaneous cerebrospinal fluid otorrhea were women. Ages ranged from 34 to 79 years. Eight patients presented with serous otitis media, and two women presented with meningitis. High-resolution computed tomography demonstrated a tegmen defect with a sensitivity of 80%. Nine tegmen defects were repaired using a transmastoid approach without recurrence. One patient with a contracted mastoid and a meningoencephalocele herniating from the tegmen tympani into the attic required the temporal craniotomy approach for definitive repair. Another patient with a tegmen tympani defect developed a recurrence of cerebrospinal fluid otorrhea 8 years after a transmastoid repair using only fascia and fibrin glue. A recurrent tegmen defect in this patient was repaired using a transmastoid approach and a multilayered closure technique. CONCLUSION: The diagnosis of spontaneous cerebrospinal fluid otorrhea requires clinical suspicion in the setting of persistent serous otitis media. High-resolution computed tomography can confirm the diagnosis. The authors' findings indicate that repair through a transmastoid approach is effective if the tegmen defect can be widely visualized. The authors advocate a multilayered closure technique. PMID- 16272937 TI - Approach design and closure techniques to minimize cerebrospinal fluid leak after cerebellopontine angle tumor surgery. AB - OBJECTIVE: The purpose of the study was to identify specific aspects of surgical approach design and closure technique aimed at reducing the incidence of cerebrospinal fluid leak after cerebellopontine angle tumor surgery. STUDY DESIGN: Retrospective case review. SETTING: Tertiary referral center. PATIENTS: All patients undergoing cerebellopontine angle tumor surgery at the study institution from January 1996 through September 2004. MAIN OUTCOME MEASURE: The presence or absence of cerebrospinal fluid leak after various surgical approaches for a wide variety of cerebellopontine angle tumors. RESULTS: Three hundred forty three patients underwent surgery for cerebellopontine angle tumors at the study institution during the study period. Tumor types in descending order of frequency were as follows: acoustic neuroma, 244; cerebellopontine angle meningiomas, 33; petroclival meningiomas, 32; foramen magnum meningiomas, 10; epidermoid tumors, 9; facial nerve tumors, 6; hemangiopericytomas, 3; schwannomas of glossopharyngeal/spinal accessory nerves, 3; and unusual internal auditory canal tumors, 3. Surgical approaches used for tumor resection included translabyrinthine, retrosigmoid, combined transpetrosal, far lateral/transcondylar, middle cranial fossa, and extended middle cranial fossa. During the nearly 8-year study period, four postoperative cerebrospinal fluid leaks were encountered, resulting in a leak rate of 1.2%. Two of these patients required surgical repair of their leaks; the other two stopped spontaneously. The authors describe specific aspects of approach design and closure that appear to have a positive impact on postoperative cerebrospinal fluid leak rates. CONCLUSION: Attention to specific aspects of surgical approach design and wound closure results in a reduced incidence of cerebrospinal fluid leak after surgery for cerebellopontine angle tumors. PMID- 16272938 TI - Office versus operating room insertion of the bone-anchored hearing aid: a comparative analysis. AB - BACKGROUND: The Bone-anchored Hearing Aid is indicated for patients with unilateral deafness and/or an irreversible conductive hearing loss. The insertion of this device is usually performed in the operating room under intravenous sedation or general anesthesia. We suggest that the placement of the abutment can occur easily and safely in the clinic setting, reducing time and costs for both the patient and the physician. STUDY DESIGN: Retrospective study. METHODS: Ten patients requiring 11 devices (1 bilateral) were implanted with the Bone-anchored Hearing Aid device in the outpatient clinic and another 8 patients were implanted in the operating room. All office procedures were done under local anesthesia; operating room procedures used either general anesthesia or intravenous sedation. Follow-up occurred over 12 months. Time required, cost of the procedure, and other logistical concerns between operating room and office-based insertion were evaluated and compared. RESULTS: When performed in the office, patient fees were reduced by 31%, and patient time investment was reduced 73%. Physician time was reduced by 50%. After office-based insertion, patients were able to return to their normal routine immediately after the procedure, and no chaperone or designated driver was required. No infection or complications occurred in either group. CONCLUSION: Bone-anchored Hearing Aid insertion in the clinic setting is safe and cost-effective, saving the patient and physician significant time and resources. PMID- 16272939 TI - Newborn hearing screening: effectiveness, importance of high-risk factors, and characteristics of infants in the neonatal intensive care unit and well-baby nursery. AB - BACKGROUND: In contrast to the recommendations of the Joint Committee on Infant Hearing, neonatal hearing screening programs are still not universally available, and many countries implement elective screening in high-risk newborns. OBJECTIVE: To assess the failure rates of neonates in hearing screening and the relative importance of risk factors for hearing impairment, both in neonatal intensive care units and in well-baby nursery neonates. The impact on cost-effectiveness is also evaluated. SUBJECTS: In the current study, 25,288 newborns were assessed; 23,574 were full-term newborns in the well-baby nursery and 1,714 neonates were in neonatal intensive care units. METHODS: All neonates had a general examination (including assessment for congenital anomalies and related history) and were assessed using transient evoked otoacoustic emissions. All newborns were older than 36 weeks at examination and thus had reliable transient evoked otoacoustic emissions. RESULTS: From the 23,574 full-term neonates in the well-baby nursery, 23,123 (98.1%) passed the test and 451 failed (1.9%). Fifty-three of the 23,574 neonates (0.2%) had a risk factor for hearing impairment; 44 (83%) passed the test and 9 failed (17%). Family history of congenital hearing loss and congenital anomalies were the most frequent risk factors for hearing loss. From the 1,714 neonates in neonatal intensive care units, 1,590 (93%) passed the test and 124 failed (7%). Two hundred thirty-two of the 1,714 neonates (14%) had a risk factor for hearing impairment; 205 (88%) passed the test and 27 failed (12%). In neonatal intensive care unit neonates, toxic levels of ototoxic drugs, mechanical ventilation for more than 24 hours, prematurity, and low birth weight were the most frequent risk factors for hearing loss. Congenital anomalies/syndromes were the most important risk factors for failing screening in both the neonatal intensive care unit and the well-baby nursery, as they showed the highest risk of failing hearing screening. The second most important factor in neonatal intensive care unit newborns was low birth weight, and the third was prematurity in relation to the possibility of failing hearing screening. CONCLUSION: The present study found 575 neonates failing hearing screening of 25,288 tested newborns (2.3%). The fact that 78% of newborns who failed hearing screening were in the well-baby nurseries further supports the necessity of universal hearing screening instead of selective screening in neonatal intensive care units, even with the obvious impact on cost-effectiveness. Even if limited funding lead to selective screening in neonatal intensive care units, this should not be applied to high risk newborns but to all neonatal intensive care unit neonates. Continuous assessment of risk factors and the related possibility of failing hearing screening are of paramount importance in designing hearing screening programs and refining the respective criteria. PMID- 16272940 TI - Are vestibular sensory cells preserved after destruction of Scarpa's ganglion? A study based on metastatic tumors of temporal bone. AB - OBJECTIVE: The contribution of nerve fibers to the maintenance of vestibular sensory cells is a controversial issue in previous studies using animals and has not yet been studied in humans. The authors investigated this issue by observing vestibular end organs in the temporal bone of three patients in whom the internal auditory canal was infiltrated with tumor cells, and Scarpa's ganglion cells showed complete degeneration. STUDY DESIGN: Retrospective case review. SETTING: University Hospital, Department of Otolaryngology. PATIENTS: Three patients with malignant metastatic temporal bone tumors. INTERVENTION: We investigated the preservative state of vestibular sensory hair cells with the Scalpa's ganglion was destructed. MAIN OUTCOME MEASURES: Maintenances of vestibular sensory hair cells. RESULTS: We found that sensory cells were intact despite the severe destruction of Scarpa's ganglion cells in two of the patients. CONCLUSION: The findings suggest that human vestibular sensory cells can be maintained for an indefinite period after denervation. PMID- 16272941 TI - Abnormal vestibular evoked myogenic potentials in the presence of normal caloric responses. AB - OBJECTIVE: Combined use of vestibular evoked myogenic potential (VEMP) and caloric response testing has enabled us to examine the function of the inferior and superior vestibular nerves separately. Although results of VEMP testing and caloric response testing have been reported for many diseases, a clinical entity showing abnormal VEMP responses but normal caloric test responses has rarely been reported. The aim of the study was to investigate clinical features of diseases showing abnormal VEMP responses with normal caloric test responses. STUDY DESIGN: Retrospective. SETTING: University hospital. PATIENTS: Eight hundred eleven patients with balance problems who had undergone both caloric response and VEMP testing were included in the study. MAIN OUTCOME MEASURES: The amplitudes and latencies of the first positive-negative peak of the VEMP (p13-n23) were measured. RESULTS: Forty of the 811 patients (5%) were found to have abnormal VEMP responses with normal caloric test responses. Clinical diagnoses of these patients were Meniere's disease (n = 12), acoustic neuroma (n = 8), sudden deafness with vertigo (n = 6), and other diseases (n = 6). Eight patients could not be diagnosed as having a disease already recognized. Clinical manifestations of these eight patients were rotatory vertigo in six patients and non-rotatory dizziness in two. None of these patients showed abnormalities other than VEMP responses on neurologic or neurotologic examinations. CONCLUSION: Apart from Meniere's disease, acoustic neuroma, and sudden deafness with vertigo, which are already known as diseases with abnormal VEMP responses but normal caloric test responses, some patients might be diagnosed as having a disease that involves only the inferior vestibular nerve region. PMID- 16272942 TI - Posturography can be used to screen for primary orthostatic tremor, a rare cause of dizziness. AB - OBJECTIVE: Primary orthostatic tremor is a rare neurologic condition of unknown origin characterized by a 10- to 20-Hz tremor in the legs while standing. Patients with primary orthostatic tremor usually complain of dizziness and unsteadiness that is relieved if they sit down or start to walk around. These patients might be referred to neurotology clinics. Previously, the only way to make the diagnosis has been by means of surface electromyographic recordings from the lower limbs during standing. The authors wanted to study whether posturography can be used to screen for primary orthostatic tremor. STUDY DESIGN: Retrospective case review. SETTING: Balance clinic at a tertiary referral center. PATIENTS: From September 2000 to August 2002, 701 patients were investigated at the authors' balance clinic. INTERVENTION: Static posturography on a force platform with 50-Hz sampling frequency. Recordings were made during 120 seconds of quiescent stance with open or closed eyes. MAIN OUTCOME MEASURE: Fast-Fourier transformation (FFT) analysis of recorded torque in the fore-to-aft direction to quantify the spectral power distribution in the range of 0 to 25 Hz. Visual inspection of power spectrum plots was performed. RESULTS: Five patients (0.7%) were identified (1 man and 4 women; mean age, 56 yr; age range, 36-73 yr) with narrow peaks at 8.5 to 18 Hz in the power spectra of their posturographic recordings. Surface electromyographic recordings during standing confirmed their diagnosis of primary orthostatic tremor. CONCLUSION: Analysis of power spectra from posturographic recordings is a simple, quick method to screen for primary orthostatic tremor. The condition might be more common than previously assumed. It is important to identify patients with primary orthostatic tremor because they might be misdiagnosed and left without treatment. PMID- 16272943 TI - Inner ear decompression sickness and mal de debarquement. AB - OBJECTIVE: To present a case series of vestibular symptoms appearing after combined sailing and diving activity, and to discuss the differential diagnosis and the workup algorithm. STUDY DESIGN: Case series. SETTING: Tertiary referral center. PATIENTS: Three patients aged 25 to 31 years suffering from unsteadiness and movement sensations after sailing and scuba diving. INTERVENTIONS: Neurotologic evaluation and recompression therapy in a hyperbaric chamber. MAIN OUTCOME MEASURES: The increasing popularity of marine sports and leisure activities has resulted in the exposure of a growing number of people to unique abnormalities not encountered under terrestrial conditions. The otolaryngologist who is involved in the care of these patients is required to diagnose and treat diving-related sinus and ear injuries such as barotrauma and decompression sickness, and also to be familiar with sailing-related disorientation syndromes such as seasickness and mal de debarquement. Treatment modalities for the various abnormalities differ significantly, and early commencement of treatment is often crucial for a successful outcome. CONCLUSION: Whenever doubt exists, recompression treatment must be instituted as soon as possible because of the potential for severe sequelae if the patient is left untreated, and because the risks involved in this therapy are minimal. PMID- 16272944 TI - Frequency dynamics shift of vestibular evoked myogenic potentials in patients with endolymphatic hydrops. AB - OBJECTIVE: To measure the frequency dynamics of the vestibular evoked myogenic potential in patients with endolymphatic hydrops. STUDY DESIGN: A prospective study. SETTING: A university hospital. SUBJECTS: The endolymphatic hydrops group consisted of 28 affected ears of patients with definite unilateral Meniere's disease and a control group of 36 ears of 20 healthy volunteers. INTERVENTIONS: Vestibular evoked myogenic potentials generated by tone bursts at 250, 500, 700, 1,000, 1,500, 2,000, and 4,000 Hz were measured in both groups. Vestibular evoked myogenic potentials were also measured after furosemide administration in six patients in the endolymphatic hydrops group. MAIN OUTCOME MEASURE: The frequency sensitivity of vestibular evoked myogenic potential, as evaluated by p13-n23 normalized amplitude. RESULTS: Peak amplitudes were noted at 500 Hz in the control group and at 1,000 Hz in the endolymphatic hydrops group. After furosemide loading, peak amplitude shifted to a lower frequency in four of six ears. CONCLUSION: The peak amplitude of vestibular evoked myogenic potentials in the endolymphatic hydrops group was at a higher frequency than in the control group. The frequency of the saccule (nu) should be proportional to radical(tau/sigma), where tau is the tension of membrane and sigma is its density. We advocate the hypothesis that the shift in frequency dynamics of vestibular evoked myogenic potential in patients with endolymphatic hydrops originates from the morphologic features of the saccule, analogous to an expanded balloon. PMID- 16272945 TI - Analysis of cortisol and other stress-related hormones in patients with Meniere's disease. AB - OBJECTIVE: To evaluate cortisol and catecholamine levels in patients with Meniere's disease. STUDY DESIGN: Prospective, controlled study. SETTING: Tertiary referral center. PATIENTS: Thirty patients with Meniere's disease and 18 healthy controls. MAIN OUTCOME MEASURES: Serum and saliva cortisol, serum cortisol after the dexamethasone suppression test, urine free cortisol, and urine catecholamines. RESULTS: The serum and saliva cortisol levels were higher in Meniere's patients compared with their control group: 440 +/- 127 (n = 28) versus 366 +/- 90 (nmol/L) (n = 18) and 17.2 +/- 6.1 (n = 18) versus 11.6 +/- 4.6 (nmol/L) (n = 9), respectively. Both differences were significant (p < 0.05). There were no dissimilarities in urine cortisol or urine catecholamines for either group. Twenty-eight Meniere's patients were divided into two subgroups, namely, high- and low-cortisol groups, using a serum cortisol level cutoff point of 465 nmol/L (median). The total Meniere's disease time and the duration of tinnitus tended to be longer in the high-cortisol group (p = 0.07, two-tailed). The total Meniere's disease time was 13.5 +/- 9.9 years in the high-cortisol group and 7.1 +/- 7.4 years in the low-cortisol group (n = 14 for both). The subgroups were matched for age, gender, and unilaterally or bilaterally affected ears. CONCLUSION: Patients with Meniere's disease have higher serum cortisol levels. It is suggested that these higher cortisol levels are rather the result than the cause of this chronic disease, because patients affected longer seem to have higher cortisol levels. The exact impact of these higher cortisol levels on the inner ear and endolymph homeostasis is yet unknown. PMID- 16272946 TI - Distortion of magnetic resonance images used in gamma knife radiosurgery treatment planning: implications for acoustic neuroma outcomes. AB - OBJECTIVE: To quantify the image distortion of our series of acoustic neuromas treated with gamma knife radiosurgery. STUDY DESIGN: Retrospective chart and digital radiographic file review with quantitative assessment of gamma knife treatment plans. SETTING: Tertiary referral center. PATIENTS: Patients undergoing gamma knife radiosurgery for the treatment of acoustic neuromas. INTERVENTION: Gamma knife radiosurgery. MAIN OUTCOME MEASURES: Gamma knife treatment plans containing magnetic resonance images were reviewed at each axial, sagittal, and coronal slice. The length of the greatest displacement of the treatment plan was measured and the volume of the treatment plan that fell outside of the internal auditory canal calculated. Known clinical measurements of audiometric, vestibular, facial, and trigeminal nerve functions were then compared with current measurements of tumor size. RESULTS: Twenty-two of the 23 patients had measurable image shifts on the axial images. The range of the image shift was 0 to 5.8 mm, with a mean shift of 1.92 +/- 1.29 mm (+/- standard deviation). Tumor volumes of the treatment plan that fell outside of the internal auditory canal ranged from 0 to 414 mm, with a mean of 90.5 mm. The mean percentage that fell outside of the internal auditory canal was 16.7% of total tumor volume (range, 2.4-77.6%). We could not draw any consistent correlations between degree of image shift and continued tumor growth or objective examination values. CONCLUSION: We have demonstrated a small but potentially significant shift in the treatment plan of gamma knife radiosurgery when based on magnetic resonance images. Although the image shift does not seem to affect the growth of the acoustic neuromas or auditory or facial nerve function, longer term follow-up is required to fully appreciate the true impact of this image shift. PMID- 16272947 TI - Treatment of glomus jugulare tumors in patients with advanced age: planned limited surgical resection followed by staged gamma knife radiosurgery: a preliminary report. AB - OBJECTIVE: To minimize treatment comorbidities in glomus jugulare tumor patients with advanced age while reducing pulsatile tinnitus and preserving or improving residual hearing using a limited middle ear/mastoid tumor resection and postoperative gamma knife radiosurgery to tumor remnants in the jugular foramen region. STUDY DESIGN: Retrospective consecutive case review of five patients. SETTING: Tertiary referral, academic medical center. PATIENTS: Patients with advanced age (mean, 69.6 yr; range, 61-78 yr) harboring symptomatic glomus jugulare tumors. INTERVENTION: All patients were treated with resection of middle ear and mastoid portions of tumor and subsequent gamma knife radiosurgery to jugular foramen portion of tumor. MAIN OUTCOME MEASURES: Length of hospitalization; hearing, pulsatile tinnitus, cranial nerve, and tumor control status. RESULTS: All patients were treated on an outpatient surgical basis without the need for blood transfusion. There were no incidents of a change in cranial nerve status (Cranial Nerves VII, IX, X, XI, and XII) in the immediate postoperative period. All patients had improvement or resolution of pulsatile tinnitus with preservation or improvement of preoperative hearing levels. Tumor volume was stable or reduced in all patients at mean follow-up of 19 months (range, 11-24 mo). Gamma knife radiosurgery (mean peripheral dose of 15 Gy) was not associated with any significant immediate or delayed complications. CONCLUSION: Short-term data reveals that staged microsurgical and radiosurgical therapy for glomus jugulare tumors in the symptomatic patient with advanced age is safe and yields favorable results regarding tinnitus, hearing, and cranial nerve status. Long-term data are needed to further evaluate the effectiveness of this treatment algorithm before extrapolating this treatment option to younger patients. PMID- 16272948 TI - Bell's palsy before Bell: Cornelis Stalpart van der Wiel's observation of Bell's palsy in 1683. AB - Bell's palsy is named after Sir Charles Bell (1774-1842), who has long been considered to be the first to describe idiopathic facial paralysis in the early 19th century. However, it was discovered that Nicolaus Anton Friedreich (1761 1836) and James Douglas (1675-1742) preceded him in the 18th century. Recently, an even earlier account of Bell's palsy was found, as observed by Cornelis Stalpart van der Wiel (1620-1702) from The Hague, The Netherlands in 1683. Because our current knowledge of the history of Bell's palsy before Bell is limited to a few documents, it is interesting to discuss Stalpart van der Wiel's description and determine its additional value for the history of Bell's palsy. It is concluded that Cornelis Stalpart van der Wiel was the first to record Bell's palsy in 1683. His manuscript provides clues for future historical research. PMID- 16272949 TI - Ossifying hemangioma of the internal auditory canal. PMID- 16272950 TI - Unilateral double vestibular schwannoma. PMID- 16272951 TI - Exostoses of the external auditory canal. PMID- 16272952 TI - Bone-conducted sound: physiological and clinical aspects. AB - OBJECTIVE: The fact that vibration of the skull causes a hearing sensation has been known since the 19th century. This mode of hearing was termed hearing by bone conduction. Although there has been more than a century of research on hearing by bone conduction, its physiology is not completely understood. Lately, new insights into the physiology of hearing by bone conduction have been reported. Knowledge of the physiology, clinical aspects, and limitations of bone conduction sound is important for clinicians dealing with hearing loss and is the purpose of this review. DATA SOURCES: The data were compiled from the published literature in the areas of clinical bone conduction hearing, bone conduction hearing aids, basic research on bone conduction physiology, and recent research on bone conduction hearing from our laboratory. CONCLUSION: Five factors contributing to bone conduction hearing have been identified: 1) sound radiated into the external ear canal, 2) middle ear ossicle inertia, 3) inertia of the cochlear fluids, 4) compression of the cochlear walls, and 5) pressure transmission from the cerebrospinal fluid. Of these five, inertia of the cochlear fluid seems most important. Bone conduction sound is believed to reflect the true cochlear function; however, certain conditions such as middle ear diseases can affect bone conduction sensitivity, but less than for air conduction. The bone conduction route can also be used for hearing aids; since the bone conduction route is less efficient than the air conduction route, bone conduction hearing aids are primarily used for hearing losses where air conduction hearing aids are contraindicated. PMID- 16272954 TI - Transcranial magnetic stimulation for tinnitus. PMID- 16272955 TI - The role of the cytochrome P450 3A5 enzyme for blood pressure regulation in the general Caucasian population. AB - Cytochrome P450 3A (CYP3A) enzymes are important for drug metabolism in gut and liver. The CYP3A5 isoenzyme is also expressed in the kidney and has been implicated in renal sodium reabsorption and blood pressure regulation. Its expression and activity is strongly linked to a polymorphism (i.e. 6986G > A). Thus, appreciable expression is found in carriers of the CYP3A5*1 (6986A) but not in homozygous carriers of the CYP3A5*3 (6986G) allele. We tested whether the presence of CYP3A5*1 affects blood pressure in Caucasian individuals who were enrolled in the Prevention of REnal and Vascular ENd stage Disease (PREVEND) study. In addition, we evaluated whether the genetic effect of CYP3A5*1 on blood pressure is modulated by sodium intake. CYP3A5*1 was found in 13.3% (901 individuals) of the cohort (6777 individuals). Diastolic blood pressure was not affected by CYP3A5*1. Overall, systolic and pulse pressure were significantly lower in carriers of CYP3A5*1, both after univariate analysis adjusted for age (P = 0.012 and P = 0.008) and in logistic regression analysis (P = 0.015 and P = 0.012). The effect on systolic blood pressure was significantly modulated by sodium intake (P = 0.038). In separate analysis according to gender, CYP3A5*1 accounted for a significant age adjusted decrease in systolic blood pressure ( 1.6 mmHg, P = 0.04) and pulse pressure (-1.2 mmHg, P = 0.04) in females but not in men. The present study demonstrates that the CYP3A5*1 allele affects systolic blood pressure and pulse pressure in the general population. Its role in hypertensive disease and potential gender differences should be investigated in further studies. PMID- 16272956 TI - CYP2A6, MAOA, DBH, DRD4, and 5HT2A genotypes, smoking behaviour and cotinine levels in 1518 UK adolescents. AB - OBJECTIVES: Smoking is a major cause of death and often initiates in adolescence. Mutations in CYP2A6 slow metabolism of nicotine to cotinine. Haploinsufficiency in adults is associated with lower cigarette consumption, lower cotinine level and higher quit rates. Other genes are also implicated in smoking behaviour. We explored smoking behaviour and cotinine levels in relation to genotypes in adolescents. METHODS: 1518 subjects from the Ten Towns Heart Health Study were genotyped for CYP2A6 alleles *1A, *1B, *2, *4, *5, *9 and *12 to classify predicted nicotine metabolism rate. DBH(rs77905), MAOA(rs1801291+VNTR), DRD4(VNTR) and 5HT2A(rs6313) were also studied. Smoking status was established by questionnaire and salivary cotinine measurement at 13-15 and 18 years. RESULTS: No significant associations were identified for DBH, MAOA, DRD4 and 5HT2A markers, with smoking status or cotinine level at either age. At age 18, haploinsufficiency (HI) for CYP2A6 was associated with a higher odds of being a current smoker compared with the *1B carriers (WT1B) (OR = 2.23 (1.16, 4.27) for current versus ex); *1A homozygotes (WT1A) were also at slightly higher risk (OR = 1.44 (1.01, 2.06)). Partial haploinsufficiency (PHI) was not associated with being a current smoker. There were no significant associations at age 13-15. PHI and HI were associated with higher cotinine levels amongst smokers at both 13-15 and at 18 years compared with WT1B and WT1A groups. CONCLUSIONS: CYP2A6 haploinsufficiency increases likelihood of continuing smoking in teenagers. We hypothesize an explanatory 'occupancy' model to explain why haploinsufficiency results in faster progression to nicotine dependence, but lower subsequent consumption. PMID- 16272957 TI - Interaction magnitude, pharmacokinetics and pharmacodynamics of ticlopidine in relation to CYP2C19 genotypic status. AB - OBJECTIVES: The aim of this study was to investigate the impact of CYP2C19 polymorphism on the extent of the interaction and on the pharmacokinetics and pharmacodynamics of ticlopidine. METHODS: Homozygous (hmEMs) and heterozygous extensive metabolizers (htEMs), and poor metabolizers (PMs, n = 6 each) took an oral dose (20 mg) of omeprazole. After a 1-week washout period, each subject received ticlopidine (200 mg) for 8 days, and ticlopidine pharmacokinetics were studied on days 1 and 7. On day 8, omeprazole was given again and its kinetic disposition was compared with that in the first dose. ADP-induced platelet aggregation was measured as a pharmacodynamic index. RESULTS: In contrast to the PMs, whose mean kinetic parameters were not altered by the repeated dosings of ticlopidine, an eight- to 10-fold increase in the mean AUC ratio of omeprazole to 5-hydroxyomeprazole was observed in both the EM groups. No significant intergenotypic differences in the pharmacokinetic parameters of ticlopidine were observed, although the accumulation ratio tended to be greater in hmEMs than in PMs (2.4 +/- 0.2 versus 1.7 +/- 0.2). A significantly positive correlation (P = 0.031) was observed between the individual percent inhibition of platelet aggregation and AUC0-24 of ticlopidine regardless of the CYP2C19 polymorphism. CONCLUSIONS: Ticlopidine is a potent inhibitor for CYP2C19 and may be associated with the phenocopy when CYPC19 substrates are co-administered to EMs. Whether and to what extent CYP2C19 would be involved in the metabolism of ticlopidine remain unanswered from the present in-vivo study. PMID- 16272958 TI - Genetic variability of CYP2B6 in populations of African and Asian origin: allele frequencies, novel functional variants, and possible implications for anti-HIV therapy with efavirenz. AB - The present study investigated CYP2B6 genetic variability by sequencing genomic DNA samples of African-American, Ghanaian, Taiwanese, Japanese and Korean subjects throughout all exons and exon-intron boundaries. The most common nonsynonymous single nucleotide polymorphisms (SNPs) were 15631G > T (Q172H) and 18053A > G (K262R, together defining allele 2B6*6), both of which had frequencies close to 50% in Ghanaians and 30% in African-Americans. These SNPs have recently been shown to affect efavirenz pharmacokinetics and response in HIV patients. Eight new missense mutations (76A > T [T26S], 83A > G [D28G], 85C > A, 86G > C [both R29T], 15618C>T [T168I], 18038G > A [D257N], 21034C > T [R336C], 21498C > A [P428T]), three new silent mutations and two new intronic SNPs defining six novel alleles (*17A and B, *18, *19, *20, *21) were identified. Heterologous expression in COS-1 cells revealed pronounced reduction in expression and/or bupropion hydroxylase activity for variants T168I, D257N, R336C and P428T, whereas the triple mutant 2B6.17 (T26S, D28G, R29T) appeared to be functionally normal. These data extend the CYP2B6 knowledge base and should be particularly relevant for anti-HIV-therapy with efavirenz. PMID- 16272959 TI - Identification and functional analysis of genetic variants of the human beta glucuronidase in a German population sample. AB - The deleterious consequences of total beta-glucuronidase deficiency, leading to symptomatic mucopolysaccharidosis type VII (MPS VII), have been firmly established. However, the question of whether sequence variations in beta glucuronidase of non-MPS VII patients affect expression of the enzyme, thereby explaining the wide inter-individual expression, has not been addressed in a systematic manner. In the present study, a population of 965 subjects were screened for enzyme activity and relevant fractions of the beta-glucuronidase gene were sequenced in those individuals belonging to the highest or lowest decile of activity. The study showed a substantial inter-individual variability of beta-glucuronidase in plasma (range 0.5-150.2 micromol/min) and confirmed the association of beta-glucuronidase activity with gender (P < 0.001), age (r = 0.218; P < 0.001) and body mass index (r = 0.311; P < 0.001). We were able to identify six beta-glucuronidase single base substitutions (-1026A > G, -72G > T, 12G > A, +7728C > T, +14 209C > T and +14 604A > G) in 193 non-MPS VII patients at a rate of one single nucleotide polymorphism (SNP) per 520 bp sequenced. The GG genotype of +14 604A > G and the CT genotype of +14 209C > T were associated with higher beta-glucuronidase activity (P < 0.05). Subsequently, reporter gene assays were carried out to elucidate the effects of the SNPs -1026A > G, -72G > T and -12G > A, and the combined genotype -1026A > G and -12G > A observed in one of the subjects. Variant -12G > A reduced the promoter activity (75%; 95% confidence interval 70-84%, P < 0.05). The present study demonstrates that three of the described SNPs influence the activity and/or expression of beta glucuronidase. Taken together, the data indicate a rather limited influence of genetic factors on inter-individual variability in beta-glucuronidase activity. PMID- 16272960 TI - Pharmacogenetics and obesity: common gene variants influence weight loss response of the norepinephrine/dopamine transporter inhibitor GW320659 in obese subjects. AB - BACKGROUND: GW320659, a highly selective neuronal norepinephrine and dopamine re uptake inhibitor, has been evaluated for the treatment of obesity. Scrutiny of the weight loss data from a phase II study (GlaxoSmithKline study OBS20001) showed a wide variation in weight loss response following GW320659 treatment and the possibility that the study population might include subgroups with enhanced weight loss response. METHODS: Pharmacogenetic analysis was performed in 191 subjects prospectively ascertained from a Phase II dose ranging study to evaluate the influence of genotype on weight loss efficacy and safety of GW320659 in obese subjects. RESULTS: Common genetic polymorphisms in the drug target (norepinephrine transporter protein 1, SLC6A2) and mechanism pathway (NMDA receptor channel NR1 subunit, GRIN1) were associated with increased weight loss following GW320659 treatment in a proportion (36%) of the study population. In the patient subgroup selected for these genotypes, GW320659 (15 mg/day) produced a significant difference in mean weight loss of 7.84 kg (SD 5.23, n = 14), compared to 2.53 kg (SD 5.17, n = 24) in the subgroup that did not possess the genotypes (P = 0.006). This subgroup also showed a highly significant weight loss response for GW320659 compared to placebo (+0.31 kg, SD 3.32, n = 16) with the same genotypes (P < 0.0001). In addition, there was no difference in placebo response between either subgroup. CONCLUSIONS: Polymorphisms in SLC6A2 and GRIN1 could be used to maximize effective obesity pharmacotherapy by norepinephrine/dopamine transporter inhibitors by identifying patients that may be predisposed to particularly good treatment weight loss response. PMID- 16272961 TI - A GSTP1 functional variant associated with cocaine dependence in a Brazilian population. AB - Cocaine dependence aetiology is complex and genetically influenced. We hypothesize that, for many users, efficient metabolism of cocaine and its toxic byproducts aids persistent cocaine use, such as that leading to dependence. The glutathione-S-transferases - in particular, GST-Pi - may be important in preventing cocaine and alcohol-induced oxidative damage. We genotyped a GST-Pi functional polymorphism (Ile105Val) in 654 male cocaine users and 572 controls from Brazil. Genotype and allele frequencies of Ile105Val differed significantly (chi = 6.74; P=0.03 and chi = 6.54; P = 0.01, respectively). Ile/Ile cocaine dependents had an OR = 1.31 (95%CI: 1.04-1.65), and Ile/Ile dependents consuming >50 units alcohol weekly an OR of 1.44 (95% CI:1.06-1.96). Population stratification was assessed and did not affect the results. These data require replication but do suggest that the high activity Ile105 GST-Pi allele may influence the aetiology and development of cocaine dependence. PMID- 16272964 TI - [Pulmonary complications of Behcet's disease and Takayasu's arteritis]. AB - INTRODUCTION: Behcet's disease and Takayasu's arteritis are both forms of large vessel vasculitis involving autoreactive T cells active against HSP 60. STATE OF THE ART: Behcet's disease is more common than Takayasu's arteritis. Pulmonary aneurysms represent the major complication of pulmonary Behcet's disease and have a poor prognosis, being associated with massive haemoptysis. In situ pulmonary artery thrombus can lead to pulmonary infarction. Superior vena cava thrombosis progresses slowly, allowing the development of a prominent collateral circulation. Vascular inflammation can spread to the mediastinum, the pleura and the lungs with diffuse pulmonary haemorrhages, bronchiolitis and organising pneumonia. Electron beam tomodensitometry and MRI are the best diagnostic techniques for assessing pulmonary vascular lesions. In Takayasu's arteritis the pulmonary arteries are less frequently involved than the aorta. Pulmonary hypertension (PHT) and lung infarcts can complicate pulmonary arterial involvement and the association of malaise, fever and weight loss with PHT is characteristic of the disease. The diagnosis is confirmed by imaging means, particularly MRI, demonstrating parietal vascular inflammation. CONCLUSION: Treatment of both conditions is based upon steroids and immunosuppression. Endovascular treatment may be used in a complementary way for aneurysms, arterial occlusion, and vena cava obstruction. However, patients often respond poorly to treatment and clinical trials using TNFalpha blockade, interferon alpha and vasodilators are in progress. PMID- 16272970 TI - [Aspergillus in the immunocompetent patient. Towards a new classification?]. PMID- 16272969 TI - [Control of peri-operative smoking: A challenge for the pneumologists]. AB - The experts conference on control of smoking in surgical patients led a synthesis of the literature which reveals among smokers 3 times more complications of the surgical site, twice more transfer in intensive care unit, an increase length of hospital stay, a 8-fold increase of the risk of non consolidation of bone, whereas quitting tobacco 6-8 weeks before the surgical procedure and until the end of the consolidation makes disappear surgical over risk related to tobacco smoke. An organization must urgently set up in all surgical units, so that the 2 millions smokers who underwent surgical procedure each year in France may be advertised and help to quit. If the mobilization of the decision makers, health care staff and patient is strong, the excepted health and financial benefit will be high and may contribute to reduce significantly the deficit of French national health insurance organization since 2006. The lung specialists have a key role to play in the implementation of these new practices. PMID- 16272971 TI - [Tracheostomy in intensive care - the debate rages on]. PMID- 16272972 TI - [Role of surgery in the management of severe haemoptysis]. PMID- 16272973 TI - [The role of tobacco centres in smoking cessation programs]. PMID- 16272974 TI - [Romanian pulmonary medicine - Quo vadis?]. PMID- 16272975 TI - [Removal of a tracheostomy in ventilator-dependent patients with neuromuscular disease]. AB - BACKGROUND: Ventilation via a tracheostomy is effective but very restricting in patients with neuromuscular disease. Return to non-invasive ventilation (NIV) is possible but this is not common practice, partly for want of standardised procedures ensuring a safe transition. METHODS: A procedure for transfer of ventilation via a tracheostomy to a mask has been developed based on the literature and local experience (feasibility of NIV, absence of laryngo-tracheal lesions, adequate leak compensation, effective cough). It has been tested in three patients with severe but stable neuromuscular disorders (chronic polyneuropathy in two cases and progressive spinal amyotrophy on one). RESULTS: The three patients were able to be extubated and established on domiciliary ventilation in 6,7 and 10 days, at the end of which all were discharged home. After 4 months in two cases and 6 months in the other no significant complications developed, the respiratory status under NIV was comparable to that previously under tracheostomy and the patients were satisfied with the change. CONCLUSION: The proposed algorithm seems to permit a rapid and safe transition from a tracheostomy to a mask. Large scale studies are needed to verify this concept and subsequently to identify within which group a similar approach may be correctly applied. PMID- 16272976 TI - [Characteristics of smokers seeking smoking cessation services: the CDT programme]. AB - INTRODUCTION: The development of smoking cessation clinics in France since 1999 has been accompanied by the setting up of an electronic database to evaluate the appropriateness of these services to the needs of smokers. The aim of this paper is to analyse the characteristics of smokers registered in the smoking cessation services national database. METHODS: A cross-sectional population-based study was conducted in 40 smoking cessation centres from 20 French regions. The study population included 14,574 smokers attending the smoking cessation centres that participated in building a national computerised database during the period 2001 2003. RESULTS: A significant proportion of the study population was female (51.4%), middle-aged (42.8 years), more highly educated (34% had received further education) and employed (68%). Almost half of the population was considered to be highly dependent on tobacco. Thirty-four percent of smokers had a past medical history of cardiovascular or lung disease. A history of depression was found in nearly one third of the population. CONCLUSIONS: Young people and individuals from deprived backgrounds were underrepresented, but smoking cessation services were being accessed by highly-dependent smokers and smokers with tobacco-related diseases services. More targeted smoking cessation strategies should be considered in order to improve access to smoking cessation services by more deprived groups. PMID- 16272977 TI - [Tracheostomy in intensive care: impact on prognosis and the influence of underlying chronic respiratory insufficiency]. AB - INTRODUCTION: Numerous uncertainties remain concerning the place of tracheostomy in intensive care. Reluctance to perform tracheostomy is common, particularly in the presence of pre-existing chronic respiratory insufficiency (CRI), but some data suggest there may be benefits. The objective of this study was to evaluate the influence of tracheostomy on mortality in both intensive care and hospital, and to study the role of pre-existing CRI. MATERIAL AND METHODS: In a retrospective study of the records of 2901 patients admitted over a period of 5 years 882 were identified who had been intubated and ventilated. 127 patients who had had tracheostomies (T+) were compared with 755 who had not (T-), and with a sub-group of T- patients (T-app) matched for severity on admission (SAPSII). RESULTS: ICU and hospital mortality were significantly less in the T+ than the T patients (28 vs 52% and 42 vs 59%) and the duration of stay was longer. This was equally true when matched for severity on admission when T+ were compared with T app (28 vs 49% and 42 vs 59%). Pre-existing CRI did not influence the outcomes of the tracheostomised patients, regardless of whether the CRI was obstructive, restrictive or neuro-muscular. CONCLUSIONS: Tracheostomy can, in certain groups of artificially ventilated patients and in certain care settings, be associated with a reduction in hospital mortality. PMID- 16272978 TI - [Interpreting carbon monoxide transfer coefficient: significance and difficulties]. AB - INTRODUCTION: Interpreting measurements of carbon monoxide can be problematic. STATE OF THE ART: The transfer factor (TLCO) is usually calculated by multiplying the decrease in alveolar CO concentration between the beginning and the end of a 10 second breath hold (KCO) by the alveolar volume (VA). Thus a reduced TLCO may occur due to either a low KCO, a reduced VA, or a combination of both. PERSPECTIVES: A careful examination of KCO and VA will usually suggest a specific explanation for a reduction in TLCO. In restrictive lung diseases from extrapulmonary origin, TLCO is low but TLCO/VA [[/INF 120% of the reference values. In interstitial lung disease, the value of TLCO/VA depends on whether the loss of lung units is diffuse or not, and probably also on the status of the microcirculation. In airflow obstruction, a low VA is caused by uneven distribution of inspired He/CO mixture within the breath-hold time. CONCLUSIONS: The transfer factor for carbon monoxide is best interpreted in terms of its components, alveolar volume and carbon monoxide transfer coefficient. PMID- 16272980 TI - [Massive haemoptysis: management and treatment. What is the role of surgery?]. AB - OBJECTIVE: To define the role of interventional radiology and surgery respectively, in the treatment of massive haemoptysis. GENERAL CONSIDERATIONS: For the management of massive haemoptysis in non-terminal pathologies an intensive care facility and a multi-disciplinary team are necessary. It is of paramount importance to identify rapidly the pulmonary or bronchial source of the bleeding. CT scanning and bronchoscopy are essential to localise the bleeding and determine its cause. Initial management. An attempt to control the initial bleeding to allow localisation of its origin and determine the treatment. TREATMENT: Bronchial or systemic embolisation and surgery are the only effective medium and long-term treatments. Embolisation achieves excellent results in bleeding from bronchial or parietal systemic arteries prior to surgery and may be the only technique possible in the presence of major co-morbidity. Surgery is necessary in the case of failure, in certain specific conditions, and in the case pulmonary artery haemorrhage from a proximal lesion. Various surgical techniques are available depending on the type of lesion encountered and the facilities for post-operative care. Emergency surgery carries a high risk and deferred surgery gives better results. CONCLUSION: The management of massive haemoptysis should be multi-disciplinary. Intensive care, respiratory and radiological diagnosis, Surgical management and interventional radiology should be combined to improve the prognosis of this grave condition. Pulmonary arterial haemorrhage from a necrotic tumour constitutes a surgical emergency and should be operated on without delay. PMID- 16272979 TI - [Genetic susceptibility to mycobacterial disease: Mendelian disorders of the interleukin-12 -interferon-gamma axis]. AB - INTRODUCTION: Environmental non tuberculous mycobacteria and Bacillus Calmette Guerin vaccines are weakly virulent mycobacteria. Nevertheless they may cause severe diseases in otherwise healthy children with no overt immunodeficiency. Parental consanguinity and familial forms are frequently observed among these patients, therefore this syndrome was named "Mendelian Susceptibility to Mycobacterial Disease". STATE OF THE ART: In the last nine years, fife genes have been found to be mutated in patients with this syndrome: IFNGR1, IFNGR2, STAT1, IL12B, IL12RB1. Allelic heterogeneity accounts for ten distinct genetic disorders. Clinical phenotype differs between patients. The spectrum of disease extends from early-onset overwhelming mycobacterial infection to adult-onset localized disease and tuberculosis. Impaired IFN-gamma-mediated immunity is the common mechanism of the disease, outlining its major role in mycobacterial immunity. PERSPECTIVES AND CONCLUSIONS: Better understanding of these disorders reveals an expanding clinical phenotype which justifies studying adult patients with pulmonary non tuberculous mycobacterial infection without known risk factors, severe BCGitis and recurrent tuberculosis. Molecular diagnosis makes it possible to introduce a specific regimen based on physiopathology. PMID- 16272981 TI - [Surgical aspects of (cardio) pulmonary transplantation]. AB - INTRODUCTION AND STATE OF THE ART: Both short and long-term outcomes following lung transplantation have improved substantially in recent years as a result of advances in the selection and management of donors, organ preservation, immunosuppressive therapy, and the treatment of infectious and malignant complications. In addition surgical techniques have evolved over time and have contributed to this increase in success rates. PERSPECTIVES AND CONCLUSIONS: This review outlines surgical aspects of lung transplantation including a historical note, techniques of lung harvesting, some anaesthetic considerations, the different transplant types and incisions, as well as anastomotic techniques and their pitfalls. PMID- 16272982 TI - [Genetics of pulmonary arterial hypertension: recent data and practical applications]. AB - INTRODUCTION: Pulmonary arterial hypertension (PAHT) is defined as an increase of mean pulmonary artery pressure above 25 mmHg at rest, or 30 mmHg on exercise, due to obliteration of small calibre pulmonary arteries by proliferation of endothelial cells and smooth muscle. Beside idiopathic PAHT and that associated with other conditions, a familial form has been identified. STATE OF THE ART: Family studies have shown an association between mutations of the BMPR2 gene and PAHT phenotypes. The products of this gene appear to be involved in vascular homeostasis and its mutations are the basis of a loss this function and, in consequence, proliferation of pulmonary vascular cells. PERSPECTIVES: Certain characteristics, such as incomplete penetrance and genetic anticipation, lead to a complex relationship between genotype and phenotype and make genetic counselling difficult. Other members of the extended family of TGF-beta receptors are implicated in the development of the Osler-Rendu syndrome, but may also be associated with the development of PAHT. CONCLUSION: Progress in genetics allows better understanding of the pathophysiology of this disease and could lead to new therapeutic possibilities. PMID- 16272984 TI - [Aspergillus bronchitis and aspergilloma treated successfully with voriconazole]. AB - INTRODUCTION: Aspergillus fumigatus is a ubiquitous soil-dwelling organism, which can cause both aspergillomas which develop in a preformed lung cavity, and aspergillus bronchitis. The two pathologies can occasionally co-exist, notably in patients with of cystic fibrosis. CASE REPORT: We describe a 57 year old patient, with diffuse bronchiectasis, who developed aspergillus bronchitis as well as an aspergilloma complicating a cavity caused by an atypical mycobacterial infection. After one month of therapy with voriconazole the aspergilloma had decreased in size and the endobronchial changes had resolved. CONCLUSION: This case report illustrates that in addition to its established role for the treatment of invasive aspergillosis, voriconazole is a promising new therapy for the treatment of aspergilloma and aspergillus bronchitis. PMID- 16272983 TI - [Invasive pulmonary aspergillosis complicating septic shock]. AB - INTRODUCTION: Invasive pulmonary aspergillosis (IPA) is an opportunistic infection with a poor prognosis, occurring primarily in patients who are severely immunocompromised. CASE REPORT: We report a case of IPA that occurred in a 37 year-old woman with no history of previous immunosuppression or significant co morbidity. She was admitted to our intensive care unit (ICU) with septic shock and multi-organ failure complicating a bacterial pneumonia. After an initial improvement, her condition deteriorated on the 10th day after admission with fever and lesions consistent with IPA seen on high-resolution computed tomography (HRCT). IPA was confirmed by isolating Aspergillus fumigatus from bronchoalveolar lavage and by a positive circulating galactomannan test (sandwich ELISA). First line therapy with voriconazole had to be stopped after 12 days due to hepatic toxicity. The patient was successfully treated with caspofungine for 2 months then itraconazole for 4 months with progressive improvement of HRCT abnormalities. Her galactomannan test became negative at 6 months. CONCLUSION: The diagnosis of IPA must be considered in critically ill patients even in the absence of underlying immunosupression where suggestive HRCT abnormalities occur in the context of septic shock and multiple organ failure. PMID- 16272985 TI - [Massive haemoptysis associated with pulmonary Strongyloides stercoralis hyperinfestation]. AB - INTRODUCTION: Pulmonary infestation with Strongyloides stercoralis is an exceptionally rare cause of haemoptysis, the diagnosis being difficult and often delayed. CASE REPORT: We report the case of a retired coal miner suffering from pneumoconiosis who presented with acute respiratory insufficiency and massive haemoptysis, with a fatal outcome, associated with pulmonary stongyloidosis. The only identified source of infestation with Strongyloides stercoralis was his period in the coal mine and the only risk factors for the hyperinfestation were a short course of systemic corticosteroid therapy and the presence of a peritoneal auricular valve. CONCLUSION: This observation illustrates the importance of a systematic search for anguillosis in ex coal miners prior to any immunosuppressant treatment in order to avoid the serious and frequently fatal form of hyperinfestation with Strongyloides stercoralis. PMID- 16272986 TI - [Pleural effusion following hepatic radiofrequency]. AB - INTRODUCTION: Surgery is the treatment of choice for primary or secondary hepatic tumours. If surgery is contra-indicated radiofrequency offers an alternative treatment capable of destroying tumours small in size and/or number. It is undertaken percutaneously, during laparotomy or endoscopically. Respiratory physicians should be familiar with certain complications. CASE REPORTS: We report two cases of pleurisy following hepatic radiofrequency. The effusions were diagnosed 15 days and one month after radiofrequency treatment for liver metastases from colonic carcinoma. The effusions were exudative and predominantly lymphocytic. The first effusion required thoracoscopy to establish the diagnosis; the second dried up following 5 pleural aspirations. CONCLUSION: Post radiofrequency pleurisy may develop late and present problems in diagnosis and treatment. PMID- 16272987 TI - [Bilateral lung cavitation and acute renal failure]. PMID- 16272988 TI - [Chondromas - a rare lung tumour]. PMID- 16272989 TI - [Survival analysis - the log rank test]. PMID- 16272990 TI - [Antithrombotic therapy for venous thromboembolism - report of the 7th Conference of The American College of Chest Physicians]. PMID- 16272991 TI - [Antithrombotic therapy for venous thromboembolism - report of the 7th Conference of The American College of Chest Physicians]. PMID- 16272994 TI - [What is important in asthmatic patients' consultations?]. PMID- 16272995 TI - [How can patients' compliance to inhalers be obtained?]. PMID- 16272996 TI - [On what criteria is the choice of an inhaler made for the treatment of asthma?]. PMID- 16272997 TI - [Novolizer: what place among inhaled treatments?]. PMID- 16272998 TI - [What are the main problems with inhaled treatment in children?]. PMID- 16272999 TI - [How should clinical practice guidelines be implemented? Conclusion]. PMID- 16273001 TI - [Pemetrexed: new multitargeted antimetabolite]. PMID- 16273002 TI - [Malignant pleural mesothelioma. Role of surgery]. PMID- 16273003 TI - [Medical treatment of malignant pleural mesothelioma. Role of pemetrexed in current treatment]. PMID- 16273004 TI - [Second-line treatment of advanced non-small-cell lung cancer: role of pemetrexed]. PMID- 16273005 TI - [Role of pemetrexed given with radiotherapy]. PMID- 16273006 TI - [First-line non-small-cell lung cancer: role of gemcitabine]. PMID- 16273007 TI - [Therapeutic strategies in non-small-cell lung carcinoma: how to optimize patient management?]. PMID- 16273008 TI - [Has survival of non-small-cell lung cancer patients been ameliorated over the course of the past 20 years?]. PMID- 16273009 TI - [Good medication usage in hospitals: results of an independent expert and multidisciplinary study of antineoplastic agents]. PMID- 16273010 TI - [New therapies, pharmaco-economic impact of lung cancerology]. PMID- 16273011 TI - Functional disorders and comorbidity. PMID- 16273012 TI - Interactive case conference: depression, dementia, or pseudodementia? PMID- 16273013 TI - An 8q21 deletion in a patient with comorbid psychosis and mental retardation. AB - Systematic investigations indicate that some of the recognized psychiatric disorders can be identified among those with mental retardation due to chromosomal abnormalities. We report a psychotic patient with mild mental retardation (intelligence quotient: 68) and minor anomalies that had a chromosomal aberration not previously described in a psychotic patient. Our patient highlights the importance of the cytogenetic study in psychiatric patients with comorbid mental retardation or minor anomalies. In addition, her psychosis symptoms may be helpful to propose a new candidate gene for psychosis. PMID- 16273014 TI - Statins and cognition: what can we learn from existing randomized trials? AB - BACKGROUND: Statins are being developed as treatments for Alzheimer's dementia based on evidence from preclinical and observational studies. However, cholesterol plays an integral role in cell membrane signal transduction and suboptimal cholesterol level could potentially impair neuronal function. Additionally, results of observational nonrandomized studies may have been affected by treatment bias. METHODS: We performed a systematic literature review in MEDLINE from January 1966 to July 2004 and included published prospective, randomized, and placebo-controlled human studies that examined the cognitive effects of statins. RESULTS: Nine studies with sample sizes ranging from 22 to 20,000 and duration of 3 weeks to 5 years, met criteria for review. Study populations and cognitive outcomes varied. Four studies were >6 months or longer. Overall, none of these studies reported finding a positive benefit for any statin on cognition in non-demented subjects although there was inconsistent evidence for acute cognitive worsening in some studies. CONCLUSION: While statins intuitively have appeal for the prevention or treatment of dementia, any conclusions about their efficacy should await more definitive evidence from on going prospective clinical trials. PMID- 16273016 TI - Pain perception in irritable bowel syndrome. AB - Irritable bowel syndrome (IBS) is a functional gastrointestinal disorder characterized by chronic abdominal discomfort or pain in the absence of detectable organic disease. IBS is common and is associated with a significant impairment in health-related quality of life. Enhanced perception of visceral stimuli ("visceral hypersensitivity") appears to be an important pathophysiological mechanism. Early IBS studies using functional brain imaging techniques suggest an alteration in central pain modulation circuits, rather than an increased sensitivity of peripheral visceral pain pathways. The frequent comorbidity with psychiatric disorders suggests the possibility of shared pathophysiological mechanisms and etiologic factors. PMID- 16273017 TI - The use of antidepressants in functional gastrointestinal disorders: new uses for old drugs. AB - Since their introduction 50 years ago, antidepressants have been used in a wide variety of settings involving gastrointestinal (GI) disease. In the 1950s, antidepressants were shown to have some efficacy for the treatment of peptic ulcer disease. This is most likely due to their antihistaminic and anticholinergic effects. Since then, more efficacious and more disease-specific treatments have become available. In the last 20 years, antidepressants have been increasingly used for the treatment of functional gastrointestinal disorders such as irritable bowel syndrome, noncardiac chest pain, and other functional GI disorders. This article will review the rationale for the use of antidepressant drugs for the treatment of functional GI disorders. The role of psychiatric comorbidity in functional GI disorders, the impact of antidepressants on GI motility and visceral sensation, and the ability of these agents to produce improvements in the global well-being and overall quality of life will be reviewed. Finally, guidelines for prescribing and barriers to a patient's acceptance of these agents will be discussed. PMID- 16273018 TI - Cognitive-behavioral treatment of irritable bowel syndrome. AB - There is increasing evidence that supports the view that irritable bowel disorder (IBS) is a disorder of brain-gut function. Cognitive-behavioral therapy (CBT) has received increased attention in light of this recent shift in the conceptualization of IBS. This review has two main aims. The first is to provide a critical review of controlled trials on CBT for IBS. The second is to discuss ways of further developing CBT interventions that are more clinically relevant and meaningful to health care providers and individuals with a diagnosis of IBS. A theme from a CBT intervention will be presented to illustrate how CBT interventions can be incorporated within a larger social context. A review of CBT for IBS lends some limited support for improvement in some IBS symptoms and associated psychosocial distress. This conclusion needs to be expressed with some caution, however, in light of many methodological shortcomings including small sample sizes, inadequate control conditions and failure to identify primary versus secondary outcome measures. In addition, future studies will need to further develop more relevant CBT protocols that more fully integrate the patient's perspective and challenge social cognitions about this stigmatized disorder. PMID- 16273019 TI - Increased prevalence of functional gastrointestinal disorders in panic disorder: clinical and theoretical implications. AB - BACKGROUND: Functional gastrointestinal disorders (FGID) are a group of disorders characterized by recurrent gastrointestinal distress for which no structural or biochemical cause can be discerned. Irritable bowel syndrome (IBS) is an FGID estimated to affect 10% to 25% of the United States population. IBS occurs in over 40% of individuals with panic disorder, and in patients with IBS, 25% to 30% have panic disorder, which has led to speculation about possible shared pathophysiology between the two. Less is known about the prevalence of other FGID in individuals with panic disorder. OBJECTIVE: The purpose of this study was to examine the prevalence of IBS and all the other FGID in patients with current Diagnostic and Statistical Manual of Mental Disorders, Fourth Edition (DSM-IV) panic disorder. INTRODUCTION: We assessed FGIDs in 73 treatment-seeking DSM-IV panic disorder patients via the Diagnostic Interview Questions for Functional Gastrointestinal Disorders and made descriptive comparisons with a large convenience sample from an already-completed United States Household Survey (USHS), which employed the same diagnostic criteria. RESULTS: The prevalence of IBS and other FGIDs in both men and women with panic disorder were substantially higher than in the USHS respondents. Women with panic disorder had significantly more functional chest pain than men, but there was no gender difference in IBS. With the exception of functional anorectal and biliary disorders, the FGID prevalences were comparatively higher in panic disorder versus the USHS respondents. DISCUSSION: This survey supports earlier reports of a high prevalence of IBS in individuals with panic disorder and also suggests that the prevalence of several other FGIDs were comparatively high as well. Methodological limitations precluded direct statistical analysis. It may be that commonly overlapping psychiatric and often-painful FGIDs, and extra-intestinal disorders increase the risk for comorbidity in already-affected individuals via shared pathophysiology. One potential model for which there is some evidence for a role in stress, panic disorder, FGIDs and several extra-intestinal functional conditions is dysregulation of corticotropin-releasing factor function. CONCLUSION: The prevalence of FGIDs in DSM-IV panic disorder was comparatively higher than in USHS respondent community sample, which used similar FGID diagnostic criteria. The cause for the apparent close association of panic disorder with FGID may represent shared pathophysiology. Increased understanding of the mechanism of the overlap may allow for improved treatment of the significant proportion of the population suffering from comorbid psychiatric and functional medical conditions. PMID- 16273022 TI - Alzheimer's disease pathways to practice: assessing diagnosis and outcome measures. PMID- 16273023 TI - The interplay of neurotransmitters in Alzheimer's disease. AB - Evidence exists for both cholinergic and glutamatergic involvement in the etiology of Alzheimer's disease. Acetylcholine (ACh), a neurotransmitter essential for processing memory and learning, is decreased in both concentration and function in patients with Alzheimer's disease. This deficit and other presynaptic cholinergic deficits, including loss of cholinergic neurons and decreased acetylcholinesterase activity, underscore the cholinergic hypothesis of Alzheimer's disease. The glutamatergic hypothesis links cognitive decline in patients with Alzheimer's to neuronal damage resulting from overactivation of N methyl-d-aspartate (NMDA) receptors by glutamate. The sustained low-level activation of NMDA receptors, which are pivotal in learning and memory, may result from deficiencies in glutamate reuptake by astroglial cells in the synaptic cleft. This article reviews the roles of ACh and glutamate in Alzheimer's disease, with particular attention given to the overlap between cholinergic and glutamatergic pathways. In addition, the potential synergy between cholinesterase inhibitors and the NMDA receptor antagonist memantine in correcting neurologic abnormalities associated with Alzheimer's disease is addressed. PMID- 16273024 TI - Formulating a clinical practice care plan for the diagnosis and assessment of Alzheimer's disease. AB - Alzheimer's disease is a devastating neurological disorder characterized by cognitive deficits, functional impairment, and often troublesome behavioral symptoms. Unfortunately, Alzheimer's disease can be difficult to diagnose at early and even moderate stages of the disease. This article outlines simple, efficient strategies for identifying patients with potential dementia. The discussion focuses on the key components of a comprehensive assessment plan, which typically involves taking the patient's history, as well as conducting cognitive testing, laboratory testing, neurologic examination, and neuroimaging to evaluate patients for Alzheimer's disease. Finally, opportunities for the improvement of care are discussed, with an emphasis on overcoming barriers to diagnosis such as time limitations and reimbursement issues. PMID- 16273025 TI - Utilizing advanced imaging and surrogate markers across the spectrum of Alzheimer's disease. AB - Alzheimer's disease is a degenerative neurological condition characterized by the presence of beta-amyloid plaques and neurofibrillary tangles in the limbic and neocortical regions of the brain. Pittsburgh Compound-B (PIB), a benzothiazole analog, has recently been found to specifically label amyloid deposits in positron emission tomography (PET) studies of the brain, opening the door for a wide range of applications related to Alzheimer's disease. In this article, data demonstrating the specificity of PIB as a PET tracer for beta-amyloid lesions are reviewed, and the potential clinical applications of PIB PET imaging is discussed. Because amyloid plaques are common even in elderly individuals who are not suffering from dementia, the primary diagnostic function of PIB PET imaging presumably would be to rule out, rather than definitively confirm, Alzheimer's diagnoses in elderly patients. Other possible uses include monitoring plaque loads in patients receiving anti-amyloid therapy for Alzheimer's disease, as well as assessing plaque formation in unaffected individuals as a means of evaluating future Alzheimer's disease. PMID- 16273026 TI - Rationalizing therapeutic approaches in Alzheimer's disease. AB - Deficits in cholinergic and glutamatergic neurotransmission have been linked to the symptomatology of Alzheimer's disease, and current therapies for Alzheimer's, including cholinesterase inhibitors (ChEIs) and the N-methyl-d-aspartate receptor antagonist memantine, have been developed to compensate for these deficits. This article reviews the results of clinical trials involving agents approved by the United States Food and Drug Administration for use in the treatment of Alzheimer's disease (namely, ChEIs for mild to moderate Alzheimer's and memantine for moderate to severe Alzheimer's). In particular, the efficacy of current monotherapy strategies in the treatment of cognitive and functional symptoms of Alzheimer's disease will be addressed. In addition, data from a clinical trial examining the use of a ChEI in combination with memantine will also be discussed, as it has been hypothesized that ChEIs and memantine may offer synergistic benefits due to their distinct mechanisms of action. PMID- 16273027 TI - Behavioral and neuropsychiatric outcomes in Alzheimer's disease. AB - Behavioral and psychological symptoms of dementia pose significant challenges in the management of patients with Alzheimer's disease. Neuropsychiatric symptoms are associated with cognitive decline, highly impaired activities of daily living, and frontal lobe pathology. Moreover, behavioral and psychological symptoms can diminish patient quality of life, increase caregiver distress, and accelerate nursing home placement. Although these symptoms are often associated with the later stages of Alzheimer's disease, a high percentage of individuals with mild cognitive impairment or mild Alzheimer's report symptoms as well. This article provides an overview of behavioral and neuropsychiatric symptoms associated with Alzheimer's disease and discusses nonpharmacologic and pharmacologic approaches to the management of such symptoms. For patients with severe behavioral and psychological symptoms of dementia, psychotropic agents may be warranted, whereas approved therapies for Alzheimer's, including cholinesterase inhibitors and the N-methyl-d-aspartate receptor antagonist memantine, may be appropriate in less severe cases. PMID- 16273028 TI - Diversity of denitrifying bacteria isolated from Daejeon Sewage Treatment Plant. AB - The diversity of the denitrifying bacterial populations in Daejeon Sewage Treatment Plant was examined using a culture-dependent approach. Of the three hundred and seventy six bacterial colonies selected randomly from agar plates, thirty-nine strains that showed denitrifying activity were selected and subjected to further analysis. According to the morphological and biochemical properties, the thirty nine isolates were divided into seven groups. This grouping was supported by an unweighted pair group method, using an arithmetic mean (UPGMA) analysis with fatty acid profiles. Restriction pattern analysis of 16S rDNA with four endonucleases (AluI, BstUI, MspI and RsaI) again revealed seven distinct groups, consistent with those defined from the morphological and biochemical properties and fatty acid profiles. Through the phylogenetic analysis using the 16S rDNA partial sequences, the main denitrifying microbial populations were found to be members of the phylum, Proteobacteria; in particular, classes Gamma proteobacteria (Aeromonas, Klebsiella and Enterobacter) and Beta proteobacteria (Acidovorax, Burkholderia and Comamonas), with Firmicutes, represented by Bacillus, also comprised a major group. PMID- 16273029 TI - Fluoroquinolone resistance and gyrA and parC mutations of Escherichia coli isolated from chicken. AB - Escherichia coli is a common inhabitant of the intestinal tracts of animals and humans. The intestines of animals also represent an ideal environment for the selection and transfer of antimicrobial resistance genes. The aim of this study was to investigate the resistance of E. coli isolated from chicken fecal samples to fluoroquinolones and to analyze the characterization of mutations in its gyrA and parC gene related resistance. One hundred and twenty-eight E. coil isolates showed a high resistance to ciprofloxacin (CIP; 60.2%), enrofloxacin (ENO; 73.4%) and norfloxacin (NOR; 60.2%). Missense mutation in gyrA was only found in the amino acid codons of Ser-83 or Asp-87. A high percentage of isolates (60.2%) showed mutations at both amino acid codons. Missense mutation in parC was found in the amino acid codon of Ser-80 or Glu-84, and seven isolates showed mutations at both amino acid codons. Isolates with a single mutation in gyrA showed minimal inhibitory concentrations (MIC) for CIP (6 years) died of malignant glioblastoma multiforme. Event-free survival at 3, 5, and 10 years is 56, 51, and 46%, respectively. Five of six patients in >CR2 or relapse at the time of transplant died. Our data should encourage the use of alternative donor transplants early in the course of disease for children with Ph+ ALL. PMID- 16273116 TI - Unrelated donor bone marrow transplantation for beta-thalassemia major: an experience from China. AB - Although related bone marrow transplantation (BMT) is effective for thalassemia, less than 30% of patients have sibling donors. Here, we report unrelated BMT in nine thalassemic children using a high-resolution HLA typing technique to identify donors. HLA mismatches between donors and recipients were 0, 1 and 2 in 2, 5 and 2 cases, respectively. The results showed that white blood cells, platelets and hemoglobin all returned to normal at various time points, and blood transfusion was eliminated from 13 to 62 days after transplantation. Full engraftment was achieved in eight patients while ABO blood types were replaced with that of donors in five of the six ABO mismatched recipients. Acute skin GVHD was found in seven patients and acute liver GVHD in one. One patient with acute intestinal GVHD eventually developed chronic GVHD. One patient died of pulmonary hemorrhage in spite of having a fully functional graft. We conclude that this is the first successful application of unrelated BMT for thalassemia major in Chinese people and that the results will certainly expand donor resources and greatly enhance the survival and quality of life of thalassemic patients. PMID- 16273117 TI - In vivo release of non-neuronal acetylcholine from the human skin as measured by dermal microdialysis: effect of botulinum toxin. AB - 1.--Acetylcholine is synthesized in the majority of non-neuronal cells, for example in human skin. In the present experiments, the in vivo release of acetylcholine was measured by dermal microdialysis. 2.--Two microdialysis membranes were inserted intradermally at the medial shank of volunteers. Physiological saline containing 1 muM neostigmine was perfused at a constant rate of 4 microl min(-1) and the effluent was collected in six subsequent 20 min periods. Acetylcholine was measured by high-pressure liquid chromatography (HPLC) combined with bioreactors and electrochemical detection. 3.--Analysis of the effluent by HPLC showed an acetylcholine peak that disappeared, when the analytical column was packed with acetylcholine-specific esterase, confirming the presence of acetylcholine. 4.--In the absence of neostigmine, 71+/-51 pmol acetylcholine (n=4) was found during a 120 min period. The amount increased to 183+/-43 pmol (n=34), when the perfusion medium contained 1 microM neostigmine. 5.--Injection of 100 MU botulinum toxin subcutaneously blocked sweating completely, but the release of acetylcholine was not affected (botulinum toxin treated skin: 116+/-70 pmol acetylcholine/120 min; untreated skin: 50+/-20 pmol; n=4). 6.--Quinine (1 mM), inhibitor of organic cation transporters, and carnitine (0.1 mM), substrate of the Na(+)-dependent carnitine transporter OCTN2, tended to reduce acetylcholine release (by 40%, not significant). 7.--Our experiments demonstrate, for the first time, the in vivo release of non-neuronal acetylcholine in human skin. Organic cation transporters are not predominantly involved in the release of non-neuronal acetylcholine from the human skin. PMID- 16273118 TI - Human colonic myocytes are involved in postischemic inflammation through ADAM17 dependent TNFalpha production. AB - The aim of this study was to identify human colonic resident cells able to initiate an inflammatory response in postischemic injury. Postischemic colonic injury, a condition relevant to various clinical settings, involves an inflammatory cascade in intestinal tissues through the recruitment of circulating inflammatory cells. However, there is no information on the nature of resident cells of the different intestinal layers able to initiate a postischemic inflammatory response. It is however an important issue in the context of a pharmacological approach of the early phase of intestinal ischemia. We reasoned that maintaining the different colonic layers as explant cultures in an oxygenated medium immediately after colonic resection, that is, after an ischemic period, would allow one to identify the resident cells able to initiate an inflammatory cascade, without interference of recruited inflammatory/immune cells. To this end, we designed an explant culture system that operationally defines three compartments in surgical specimens of the human colon, based on the microdissected layers, that is, mucosa, submucosa (containing muscularis mucosae) and muscularis propria. To validate the results obtained in explant cultures in the clinical setting of ischemic colitis, eight cases of sigmoid volvulus were examined. Only the myocytes-containing explants produced tumor necrosis factor alpha (TNFalpha), via an ADAM17 (a disintegrin and metalloproteinase-17) dependent pathway, as shown by the abrogation of TNFalpha production by the inhibitor Tapi-2. Immunofluorescence studies identified nonvascular and vascular myocytes as resident cells coexpressing TNFalpha and ADAM17, both in our postischemic explant system and in surgical specimens from ischemic colitis patients. Finally, time-course experiments on explanted tissues showed that TNFalpha production by myocytes was an early event triggered by a postischemic oxidative stress involving nuclear factor kappa B (NF-kappaB). In conclusion, this study identifies human intestinal myocytes as resident cells able to initiate an inflammatory reaction through TNFalpha production in postischemic conditions, and delineates two points of control in TNFalpha production, NF kappaB and ADAM17, which can be targeted by pharmacological manipulation. PMID- 16273119 TI - Angiotensin II and angiotensin II receptor blocker modulate the arrhythmogenic activity of pulmonary veins. AB - Angiotensin II receptor blockers (AIIRBs) have been shown to prevent atrial fibrillation. The pulmonary veins (PVs) are the most important focus for the generation of atrial fibrillation. The aim of this study was to evaluate whether angiotensin II or AIIRB may change the arrhythmogenic activity of the PVs. Conventional microelectrodes and whole-cell patch clamps were used to investigate the action potentials (APs) and ionic currents in isolated rabbit PV tissue and single cardiomyocytes before and after administering angiotensin II or losartan (AIIRB). In the tissue preparations, angiotensin II induced delayed after depolarizations (1, 10, and 100 nM) and accelerated the automatic rhythm (10 and 100 nM). Angiotensin II (100 nM) prolonged the AP duration and increased the contractile force (10 and 100 nM). Losartan (1 and 10 microM) inhibited the automatic rhythm. Losartan (10 microM) prolonged the AP duration and reduced the contractile force (1 and 10 microM). Angiotensin II reduced the transient outward potassium current (I(to)) but increased the L-type calcium, delayed rectifier potassium (I(K)), transient inward (I(ti)), pacemaker, and Na(+)-Ca(2+) exchanger (NCX) currents in the PV cardiomyocytes. Losartan decreased the I(to), I(K), I(ti), and NCX currents. In conclusion, angiotensin II and AIIRB modulate the PV electrical activity, which may play a role in the pathophysiology of atrial fibrillation. PMID- 16273120 TI - In vitro and in vivo pharmacological characterization of the novel UT receptor ligand [Pen5,DTrp7,Dab8]urotensin II(4-11) (UFP-803). AB - The novel urotensin-II (U-II) receptor (UT) ligand, [Pen(5),DTrp(7),Dab(8)]U-II(4 11) (UFP-803), was pharmacologically evaluated and compared with urantide in in vitro and in vivo assays. In the rat isolated aorta, UFP-803 was inactive alone but, concentration dependently, displaced the contractile response to U-II to the right, revealing a competitive type of antagonism and a pA(2) value of 7.46. In the FLIPR [Ca(2+)](i) assay, performed at room temperature in HEK293(hUT) and HEK293(rUT) cells, U-II increased [Ca(2+)](i) with pEC(50) values of 8.11 and 8.48. Urantide and UFP-803 were inactive as agonists, but antagonized the actions of U-II by reducing, in a concentration-dependent manner, the agonist maximal effects with apparent pK(B) values in the range of 8.45-9.05. In a separate series of experiments performed at 37 degrees C using a cuvette-based [Ca(2+)](i) assay and CHO(hUT) cells, urantide mimicked the [Ca(2+)](i) stimulatory effect of U-II with an intrinsic activity (alpha) of 0.80, while UFP-803 displayed a small (alpha=0.21) but consistent residual agonist activity. When the same experiments were repeated at 22 degrees C (a temperature similar to that in FLIPR experiments), urantide displayed a very small intrinsic activity (alpha=0.11) and UFP-803 was completely inactive as an agonist. In vivo in mice, UFP-803 (10 nmol kg(-1)) antagonized U-II (1 nmol kg(-1))-induced increase in plasma extravasation in various vascular beds, while being inactive alone. In conclusion, UFP-803 is a potent UT receptor ligand which displays competitive/noncompetitive antagonist behavior depending on the assay. While UFP-803 is less potent than urantide, it displayed reduced residual agonist activity and as such may be a useful pharmacological tool. PMID- 16273121 TI - Protein S modulates the anticoagulant action of recombinant human activated protein C: a comparison between neonates and adults. AB - Recombinant human-activated protein C (rhAPC, Drotrecogin alpha (activated), Xigris) has been shown to reduce organ damage and decrease mortality in severe sepsis. Since protein S (PS) serves as a potentiating cofactor of activated protein C and since PS levels are low in neonatal plasma, we hypothesized that the anticoagulant effect of rhAPC would be decreased in cord plasma compared to adult plasma. We demonstrate that the anticoagulant action of 0.3 microg ml(-1) rhAPC (5 nmol l(-1)) was decreased in cord plasma compared to adult plasma, and dose dependently increased in cord plasma in the presence of increasing activities of PS. Correspondingly, the anticoagulant action of rhAPC decreased in adult plasma in the presence of decreasing activities of PS. The low anticoagulant action of rhAPC in cord compared to adult plasma is attributable to low neonatal levels of PS, and as previously shown, to low neonatal levels of TFPI and AT. Our laboratory experiments do not allow definite conclusions for clinical situations. However, we speculate that the anticoagulant efficacy of rhAPC is impaired in neonates and in clinical situations associated with consumption and/or inhibition of PS, AT, and TFPI, such as severe sepsis. PMID- 16273123 TI - Direct inhibition of the pacemaker (If) current in rabbit sinoatrial node cells by genistein. AB - Genistein is a tyrosine kinase inhibitor which interferes with the activity of several ionic channels either by altering modulatory phosphorylating processes or by direct binding. In whole-cell conditions, genistein induces a partial inhibition of the pacemaker (I(f)) current recorded in cardiac sinoatrial and ventricular myocytes. We investigated the mechanism of action of genistein (50 microM) on the I(f) current in whole-cell, cell-attached, and inside-out configurations, and the measured fractional inhibitions were similar: 26.6, 27.2, and 33.6%, respectively. When ATP was removed from the whole-cell pipette solution no differences were revealed in the effect of the drug when compared to metabolically active cells. Genistein fully maintained its blocking ability even when herbimycin, a tyrosine kinase inhibitor, was added to the whole-cell ATP free pipette solution. Genistein-induced block was independent of the gating state of the channel and did not display voltage or current dependence; this independence distinguishes genistein from all other f-channel blockers. When inside-out experiments were performed to test for a direct interaction with the channel, genistein, superfused on the intracellular side of the membrane, decreased the maximal I(f) conductance, and slightly shifted the current activation curve to the left. Furthermore, the effect of genistein was independent of cAMP modulation. We conclude that, in addition to its tyrosine kinase-inhibitory properties, genistein also blocks I(f) by directly interacting with the channel, and thus cannot be considered a valuable pharmacological tool to investigate phosphorylation-dependent modulatory pathways of the I(f) current and of cardiac rhythm. PMID- 16273125 TI - The role of Na+/H+ exchanger in serotonin secretion from porcine blood platelets. AB - This study was undertaken to evaluate whether a link exists between the activation of protein kinase C (PKC), operation of Na(+)/H(+) exchanger (NHE), cell swelling and serotonin (5-HT) secretion in porcine platelets. Activation of platelets by thrombin or phorbol 12-myristate 13-acetate (PMA), a PKC activator, initiated a rapid rise in the activity of Na(+)/H(+) exchanger and secretion of 5 HT. Both thrombin- and PMA-evoked activation of Na(+)/H(+) exchanger was less pronounced in the presence of ethyl-isopropyl-amiloride (EIPA), an NHE inhibitor, and by GF 109203X, a PKC inhibitor. Monensin (simulating the action of NHE) caused a dose-dependent release of 5-HT that was not abolished by GF 109203X or EGTA. Lack of Na(+) in the suspending medium reduced thrombin-, PMA-, and monensin-evoked 5-HT secretion. GF 109203X nearly completely inhibited 5-HT release induced by PMA-, partly that induced by thrombin, and had no effect on 5 HT release induced by monensin. EIPA partly inhibited 5-HT release induced by thrombin and nearly totally that evoked by PMA. Electronic cell sizing measurements showed an increase in mean platelet volume upon treatment of cells with monensin, PMA or thrombin. The PMA- and thrombin-evoked rise in mean platelet volume was strongly reduced in the presence of EIPA. As judged by optical swelling assay monensin and PMA produced a rapid rise in platelet volume. The swelling elicited by PMA was inhibited by EIPA and its kinetics was similar to that observed in the presence of monensin. Hypoosmotically evoked platelet swelling did not affect platelet aggregation but significantly potentiated thrombin-evoked release of 5-HT and ATP. Taken together, these results show that in porcine platelets PKC may promote 5-HT secretion through the activation of NHE. It is hypothesized that enhanced Na(+)/H(+) antiport may result in a rise in cell membrane tension (due to cell swelling) which in turn facilitates fusion of secretory granules with the plasma membrane leading to 5-HT secretion. PMID- 16273122 TI - Long-term exposure to the new nicotinic antagonist 1,2-bisN-cytisinylethane upregulates nicotinic receptor subtypes of SH-SY5Y human neuroblastoma cells. AB - Nicotinic drug treatment can affect the expression of neuronal nicotinic acetylcholine receptors (nAChR) both in vivo and in vitro through molecular mechanisms not fully understood. The present study investigated the effect of the novel cytisine dimer 1,2-bisN-cytisinylethane (CC4) on nAChR natively expressed by SH-SY5Y neuroblastoma cells in culture. CC4 lacked the agonist properties of cytisine and was a potent antagonist (IC50=220 nM) on nAChRs. Chronic treatment of SH-SY5Y cells with 1 mM CC4 for 48 h increased the expression of 3H epibatidine (3H-Epi; 3-4-fold) or 125I-alpha-bungarotoxin (125I-alphaBgtx; 1.2 fold) sensitive receptors present on the cell membrane and in the intracellular pool. Comparable data were obtained with nicotine or cytisine, but not with carbamylcholine, d-tubocurarine, di-hydro-beta-erythroidine or hexametonium. Immunoprecipitation and immunopurification studies showed that the increase in 3H Epi-binding receptors was due to the enhanced expression of alpha3beta2 and alpha3beta2beta4 subtypes without changes in subunit mRNA transcription or receptor half-life. The upregulation was not dependent on agonist/antagonist properties of the drugs, and did not concern muscarinic or serotonin receptors. Whole-cell patch clamp analysis of CC4-treated cells demonstrated larger nicotine evoked inward currents with augmented sensitivity to the blockers alpha-conotoxin MII or methyllycaconitine. In conclusion, chronic treatment with CC4 increased the number of nAChRs containing beta2 and alpha7 subunits on the plasma membrane, where they were functionally active. In the case of beta2-containing receptors, we propose that CC4, by binding to intracellular receptors, triggered a conformational reorganisation of intracellular subunits that stimulated preferential assembly and membrane-directed trafficking of beta2-containing receptor subtypes.. PMID- 16273126 TI - The HPV16 E2 transcriptional regulator mode of action depends on the physical state of the viral genome. AB - Human papillomavirus (HPV) infection is a major risk factor for the development of cervical cancer. The HPV-induced immortalization of epithelial cell usually requires integration of the viral DNA into the host cell genome. The integration event causes disruption of the E2 gene and this is followed by overexpression of the E6 and E7 oncoproteins. The E2 protein is a transcription factor that regulates expression of the E6 and E7 oncoproteins by binding to four sites within the viral long control region. We used an in vitro cell culture model to explore the role of the E2 protein in the transcriptional control of the HPV16 long control region. Employing transient and stable transfection experiments we simulated the episomal and integrated states of the viral genome, respectively. We show that the E2 protein up-regulates E6/E7 transcription from episomal DNA but represses it in the case of integrated DNA. The activator function of the E2 protein seems to counteract the repressive chromatin structure formed over episomal DNA. Steroid hormones and retinol also modulate oncogene transcription differently depending on the physical structure of the viral DNA. Our data suggest regulatory mechanisms involving interactions between the E2 protein and nuclear hormone receptors. PMID- 16273127 TI - Thermostable Pyrococcus woesei beta-D-galactosidase--high level expression, purification and biochemical properties. AB - The gene encoding beta-D-galactosidase from Pyrococcus woesei was PCR amplified, cloned, expressed in Escherichia coli under the control of an inducible T7 promoter, purified and characterized. The expression system was developed by the construction of recombinant plasmid, based on the high copy number pUET1 vector, giving four times more efficient expression of P. woesei beta-D-galactosidase (20 mg of enzyme from 1 liter of culture) than that obtained from a previously constructed one. The recombinant enzymes were purified in a two-step procedure: double heat-denaturation of E. coli cell proteins and affinity chromatography on p-aminobenzyl 1-thio-beta-D-galactopyranoside-agarose. To achieve efficient purification of P. woesei beta-D-galactosidase by immobilized metal-ion affinity chromatography (IMAC), a His-tag was placed either at the N- or the C-terminal of the coding sequence. The obtained fusion proteins revealed the same specific activity of approximately 5400 U/mg, which was 10 times lower than the wild-type beta-D-galactosidase (51100 U/mg). The activity of P. woesei beta-D-galactosidase was enhanced by thiol compounds, Mg(2+) ions and D-galactose, and was inhibited by heavy metal ions and D-glucose, while Ca(2+) ions had no effect. PMID- 16273128 TI - New spectrophotometric methods for the determination of nifedipine in pharmaceutical formulations. AB - Two simple, sensitive and economical spectrophotometric methods were developed for the determination of nifedipine in pharmaceutical formulations. Method A is based on the reaction of the nitro group of the drug with potassium hydroxide in dimethyl sulphoxide (DMSO) medium to form a coloured product, which absorbs maximally at 430 nm. Method B uses oxidation of the drug with ammonium molybdate and subsequently reduced molybdenum blue is measured at 830 nm. Beer's law is obeyed in the concentration range of 5.0-50.0 and 2.5-45.0 microg ml(-1) with methods A and B, respectively. Both methods have been successfully applied for the assay of the drug in pharmaceutical formulations. No interference was observed from common pharmaceutical adjuvants. The reliability and the performance of the proposed methods are established by point and interval hypothesis tests and through recovery studies. PMID- 16273129 TI - The effect of Topotecan on oxidative stress in MCF-7 human breast cancer cell line. AB - PURPOSE: Topotecan, a semisynthetic water-soluble derivative of camptothecin exerts its cytotoxic effect by inhibiting topoisomerase I and causes double strand DNA breaks which inhibit DNA function and ultimately lead to cell death. In previous studies it was shown that camptothecin causes ROS formation. The aim of this study was to investigate if Topotecan like camptotecin causes oxidative stress in MCF-7 human breast cancer cell line. Determining the oxidant effect of Topotecan may elucidate a possible alternative mechanism for its cytotoxicity. EXPERIMENTAL DESIGN: MCF-7 cells were cultured and exposed to Topotecan for 24 h at 37 degrees C. The viability of the cells (% of control) was measured using the colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. Lipid peroxidation (TBARS), protein oxidation (carbonyl content), sulfhydryl, glutathione (GSH) levels, superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) activities were determined in MCF-7 cells with and without Topotecan incubation. RESULTS: We found the IC(50) concentration of Topotecan as 0.218 microM in MCF-7 cells. This concentration of Topotecan was used in the incubations of the cells. Our data indicated increased oxidative status, as revealed by increased lipid peroxidation and protein oxidation, and decreased GSH and sulfhydryl levels in MCF-7 cells exposed to Topotecan compared to control cells. In contrast, there was a slight increase in SOD and a significant increase in GPx and catalase activity in MCF-7 cells incubated with Topotecan compared to the control. CONCLUSIONS: These results support our hypothesis that Topotecan increases oxidative stress in MCF-7 cells. PMID- 16273130 TI - Combined radiation and suicide gene therapy for Nasopharyngeal Carcinoma: in vitro and in vivo models. AB - Nasopharyngeal carcinoma (NPC) cells are highly radiosensitive. Historically, the main modality for NPC has been radiotherapy. Now, to explore a novel and more effective approach to NPC therapy, a combined strategy of suicide gene and radiation was developed. The yeast CDglyTK (yCDglyTK) gene controlled by a synthetic CMV-enhanced Egr-1 promoter (CE) was constructed whose expression in NPC CNE-2 cells was detected by Western Blot. The cytotoxicity of the combined radio-gene therapy was assayed by MTT. The conversion rate of 5-FC to 5-FU in vitro and the bio-distribution of 5-FU in vivo were determined by HPLC. An animal study in which yCDglyTK-expressing CNE-2 tumors were treated with 5-FC and radiation was also conducted. The results reveal that yCDglyTK is expressed in CNE-2 cells, the CEyCDglyTK/5-FC system has a potent anti-tumor action in NPC, and the radio-sensitization of Egr-1 promoter plays a key role in the killing of CNE-2 cells and in the conversion of 5-FC to 5-FU. Moreover, the treated tumors regressed significantly, and a significant difference of tumor volumes was observed between the CEyCDglyTK+5-FC+radiation group and the other four groups (P<0.05). The results also showed that suicide gene therapy and radiation have a synergic anti-tumor effect on NPC, and the combined strategy of radio-gene therapy is of great potential as a substitute for the traditional method, radiation alone, in NPC therapies. PMID- 16273131 TI - Determination of promethazine hydrochloride in pharmaceuticals by capillary isotachophoresis. AB - Capillary isotachophoresis (ITP) in cationic regime of the separation with conductometric detection has been used for the separation and determination of promethazine hydrochloride (PRO) in commercial mass-produced pharmaceutical preparations. Several electrolyte systems of different compositions and pH were examined and the optimized ITP electrolyte system consisted of 10 mmol/l of potassium acetate adjusted to pH 4.8 with acetic acid as the leading electrolyte with electroosmotic flow (EOF) suppressing additive, 0.2% (w/v) methylhydroxyethylcellulose (m-HEC), and 5 mmol/l of beta-alanine as the terminating electrolyte. The proposed electrophoretic method was successfully validated. It was convenient for the sensitive, simple, rapid, and highly reproducible assay of promethazine. The calibration graph relating the ITP zone length to concentration of the analyte was rectilinear in the range of 40-200 mg/l of the drug standard, with a coefficient of determination r(2)=0.9992. The relative standard deviation (RSD) was 1.12% (n=6) when determining 100 mg/l of PRO in standard sample. Good quantitation was obtained in short analysis time (a single analysis took 6 min). The recoveries of drug from samples were found to be 97.22% (tablets), 99.72% (injections), and 99.14% (syrup). The minimal sample pretreatment and low running cost make the proposed ITP method a good alternative to commonly used analytical methods. PMID- 16273132 TI - Evaluation of antidiarrheal potential of trichodesma indicum root extract in rats. AB - Trichodesma indicum is found as a weed throughout the greater part of India. The decoction of the root is used for diarrhea, dysentery, and fever in Indian traditional medicine. However, the traditional claims need to be validated by suitable experimental animal models. Therefore, this study was undertaken to evaluate its antidiarrheal potential in several animal models. The extract significantly inhibited the castor oil-induced diarrhea and decreased propulsion of charcoal meal through the gastrointestinal tract; it also reduced the castor oil-induced small intestinal fluid accumulation (enteropooling). The ethanol extract of T. indicum root has significant antidiarrheal activity and substantiates the use of this herbal remedy as a nonspecific treatment for diarrhea in folk medicine. PMID- 16273133 TI - Neuropharmacokinetic characterization of lamotrigine after its acute administration to rats. AB - The purpose of this study is to characterize the neuropharmacokinetics of lamotrigine following a single intraperitoneal dose. Adult male Wistar rats were given lamotrigine dose of 5, 10, or 20 mg/kg. Blood and brain samples were obtained at predetermined times over 120 h and analyzed by HPLC. The overall characteristics of plasma curves were determined by noncompartmental analysis with WINNONLIN. The kinetic characterization of lamotrigine distribution between plasma and brain was performed by indirect numerical deconvolution with MULTI(FILT). A linear disposition kinetics was observed within 5-20 mg/kg. The lamotrigine concentrations in brain homogenate were approx. twofold higher than in plasma. The following pharmacokinetic parameters were obtained for lamotrigine 5, 10, and 20 mg/kg, respectively: clearance of distribution from plasma to brain normalized with the volume of the brain, CL/V(h(-1)) = 4.64, 2.47, 2.40; brain-to plasma partition coefficient, P = 0.40, 0.37, 0.34; first-order transfer rate constant from the brain to the plasma, K(h(-1)) = 11.68, 6.68, 5.96; single-pass mean transit time in the brain, MTT(h) = 0.086, 0.150, 0.168. These results indicate that lamotrigine plasma levels may be good indicators of lamotrigine levels in the brain and that higher response intensities could be expected with higher doses of lamotrigine, since efficacious concentrations are maintained for a longer period. PMID- 16273134 TI - Antiinflammatory, analgesic and hypoglycemic effects of Mangifera indica Linn. (Anacardiaceae) stem-bark aqueous extract. AB - Previous studies in our laboratories and elsewhere have shown that some members of Anacardiaceae family possess antiinflammatory, analgesic and hypoglycemic effects in man and mammalian experimental animals. The present study was, therefore, undertaken to examine the antiinflammatory, analgesic and antidiabetic properties of the stem-bark aqueous extract of Mangifera indica Linn., M. indica a member of the Anacardiaceae family, in rats and mice. The stem-bark powder of M. indica was Soxhlet extracted with distilled water and used. The analgesic effect of the plant's extract was evaluated by the hot-plate and acetic acid test models of pain in mice, while the antiinflammatory and antidiabetic effects of the stem-bark extract were investigated in rats, using fresh egg albumin-induced paw edema, and streptozotocin (STZ)-induced diabetes mellitus, respectively. Morphine (MPN, 10 mg/kg i.p.), diclofenac (DIC, 100 mg/kg i.p.), and chlorpropamide (250 mg/kg p.o.) were used respectively as reference analgesic, antiinflammatory, and hypoglycemic agents for comparison. M. indica stem-bark aqueous extract (MIE, 50-800 mg/kg i.p.) produced dose-dependent and significant (p<0.05-0.001) analgesic effects against thermally and chemically induced nociceptive pain stimuli in mice. MIE (50-800 mg/kg i.p.) also significantly (p<0.05-0.001) inhibited fresh egg albumin-induced paw edema, and caused significant (p<0.05-0.001) hypoglycemic effects in rats. It is suggested that the analgesic effects of MIE (50-800 mg/kg i.p.) may be peripherally and centrally mediated. The different chemical constituents of the plant, especially the polyphenolics, flavonoids, triterpenoids, mangiferin, and other chemical compounds present in the plant may be involved in the observed antiinflammatory, analgesic, and hypoglycemic effects of the plant's extract. However, the results of this experimental animal study lend pharmacological credence to the suggested folkloric uses of the plant in the management and control of painful, arthritic and other inflammatory conditions, as well as in the management of adult-onset type 2 diabetes mellitus in some rural African communities. PMID- 16273135 TI - Pharmacodynamic equivalence study of CFC-free and CFC-containing procaterol hydrochloride metered-dose inhalers. AB - Equivalence between a CFC-free procaterol hydrochloride metered-dose inhaler using HFA-227 as a propellant (Meptin [HFA]) and a CFC-containing procaterol hydrochloride metered-dose inhaler (Meptin [CFC]) was assessed in 28 patients with bronchial asthma. The study was conducted in a randomized, double-dummy, double-blind crossover manner, using forced expiratory volume in the first second (FEV1) as an index of bronchodilatory effect. In Period I, the patients received 20 microg of either Meptin [HFA] or Meptin [CFC] and then crossed over in Period II after a washout interval of 3-28 days. Pharmacodynamic equivalence was assessed using AUC (FEV1)/h and peak FEV1 as indices, and the data was analyzed by analysis of variance. Factors used for the analysis were the treatment group and/or carryover effect, patients within each group, period, and treatment. The 90% confidence intervals for the differences between the two treatments were 0.0507 to 0.0039 (L) for mean AUC (FEV1)/h and -0.056 to 0.026 (L) for mean peak FEV1, both within the acceptance criteria of -0.15 to 0.15 (L). Meptin [HFA] was therefore assessed as being equivalent to the current Meptin [CFC]. PMID- 16273136 TI - Cytochrome P450 3A: genetic polymorphisms and inter-ethnic differences. AB - Ethnicity is a demographic variable that plays an important role in interindividual variability of drug metabolism and response. The genetic variations of drug-metabolizing enzymes exhibiting interindividual differences of drug metabolism also show differences between populations. The reason for this is that the frequency of a polymorphism is found to differ between populations. The other reason is that different variants are seen in different populations. Most drugs are biotransformed in the body by cytochrome P450. The CYP3A isozymes are responsible for the metabolism of 50-60% of all currently prescribed drugs. Studies have shown that there is variability in CYP3A activity and also inter ethnic differences in CYP3A-mediated drug metabolism. The purpose of this review is to focus on the genetic polymorphism and ethnic variations in CYP3A-mediated oxidative drug metabolism. PMID- 16273137 TI - Gateways to clinical trials. AB - Gateways to Clinical Trials is a guide to the most recent clinical trials in current literature and congresses. The data in the following tables have been retrieved from the Clinical Trials Knowledge Area of Prous Science Integrity, the drug discovery and development portal, http://integrity.prous.com. This issue focuses on the following selection of drugs: (-)-Epigallocatechin gallate, (Z)-4 hydroxytamoxifen; Ad.muIFN-beta AD-237, adalimumab, adefovir dipivoxil, agalsidase alfa, alemtuzumab, almotriptan, ALVAC vCP1452, alvimopan hydrate, ambrisentan, anakinra, anti-IFN-gamma MAb; Bimatoprost, BMS-188797, BMS-214662, bortezomib, bosentan, bovine lactoferrin; Caffeine, canertinib dihydrochloride, canfosfamide hydrochloride, cannabidiol, caspofungin acetate, cetuximab, cH36, ChimeriVax-JE, ciclesonide, cilansetron, cinacalcet hydrochloride, clopidogrel, CpG-7909, Cypher; Daptomycin, darbepoetin alfa, darifenacin hydrobromide, decitabine, denufosol tetrasodium, Dexamet, diindolemethane, drotrecogin alfa (activated), duloxetine hydrochloride, DX-9065a; E-7010, edaravone, efalizumab, eicosapentaenoic acid/docosahexaenoic acid, elacridar, eletriptan, emtricitabine, epratuzumab, erlotinib hydrochloride, ertapenem sodium, eszopiclone, everolimus, ezetimibe; Fludarabine, fondaparinux sodium; gamma-Hydroxybutyrate sodium, gavestinel sodium, gefitinib, granisetron-Biochronomer; Human Albumin, human insulin; Imatinib mesylate, indiplon, interleukin-2 XL, isatoribine, ISS-1018, i.v. gamma-globulin, ivabradine hydrochloride, ixabepilone; Lanthanum carbonate, L-arginine hydrochloride, liposomal doxorubicin, LY-450139; Magnesium sulfate, melatonin, motexafin gadolinium, mycophenolic acid sodium salt; Natalizumab, nesiritide, niacin/lovastatin; OGX-011, olmesartan medoxomil, omalizumab, ospemifene; PACAP38, panitumumab, parathyroid hormone (human recombinant), parecoxib sodium, patupilone, pegfilgrastim, peginterferon alfa-2a, peginterferon alfa-2b, peginterferon alfa-2b/ribavirin, pemetrexed disodium, pimecrolimus, pirfenidone, posaconazole, prasterone, pregabalin; R-112, ramelteon, ranolazine, rasagiline mesilate, rebimastat, roflumilast, rosuvastatin calcium, rotigotine hydrochloride, rupatadine fumarate; S-3013, S-3304, semustine, sitaxsentan sodium, St. John's Wort extract; Tadalafil, tamoxifen, Taxus, Tc-99m-EDDA-HYNIC TOC, TH-9507, tiotropium bromide, tipifarnib, tocilizumab, tolvaptan, torcetrapib, TR-14035, tramadol hydrochloride/acetaminophen, treprostinil diethanolamine, troglitazone, troxacitabine; Valdecoxib, valganciclovir hydrochloride, vandetanib, vardenafil hydrochloride hydrate, VAS-991, veglin, vinflunine, voriconazole; White sweet potato extract; Ximelagatran. PMID- 16273138 TI - Organic surfaces excited by low-energy ions: atomic collisions, molecular desorption and buckminsterfullerenes. AB - This article reviews the recent progress in the understanding of kiloelectronvolt particle interactions with organic solids, including atomic displacements in a light organic medium, vibrational excitation and desorption of fragments and entire molecules. This new insight is the result of a combination of theoretical and experimental approaches, essentially molecular dynamics (MD) simulations and secondary ion mass spectrometry (SIMS). Classical MD simulations provide us with a detailed microscopic view of the processes occurring in the bombarded target, from the collision cascade specifics to the scenarios of molecular emission. Time of-flight SIMS measures the mass and energy distributions of sputtered ionized fragments and molecular species, a precious source of information concerning their formation, desorption, ionization and delayed unimolecular dissociation in the gas phase. The mechanisms of energy transfer and sputtering are compared for bulk molecular solids, organic overlayers on metal and large molecules embedded in a low-molecular weight matrix. These comparisons help understand some of the beneficial effects of metal substrates and matrices for the analysis of molecules by SIMS. In parallel, I briefly describe the distinct ionization channels of molecules sputtered from organic solids and overlayers. The specific processes induced by polyatomic projectile bombardment, especially fullerenes, are discussed on the basis of new measurements and calculations. Finally, the perspective addresses the state-of-the-art and potential developments in the fields of surface modification and analysis of organic materials by kiloelectronvolt ion beams. PMID- 16273139 TI - Position-dependent energy of molecules in nano-confined water. AB - Real time decrease in the energy (or enthalpy) measured during confinement of controlled amounts of water in 2 nm radius pores of Vycor shows that exothermic transfer of bulk water to nanopores via the vapour-phase occurred in two stages. In the first stage, at saturation pressure, H2O molecules from the vapour rapidly accumulated in the nanopore channels near the Vycor surface. In the second, at vapour pressure below saturation, the accumulation rate abruptly decreased and water (slowly) diffused and redistributed in the nanopore channels until the vapour pressure equilibrium was attained. The energy decrease per H2O molecule was highest, 14.5 kJ mol(-1), at low amounts when the pore-wall was incompletely covered by H2O. This value approached zero at higher amounts when pores were gradually filled. The results show that the vibrational and configurational contributions to the energy of H2O molecules depend upon their position in the nanopore and these contributions approach their bulk water values at high water concentration, but do not attain those values for completely filled pores. PMID- 16273140 TI - Translational self diffusion anisotropy in the smectic A phase measured by a static fringe field gradient 1H NMR diffusometry approach. AB - Self diffusion along the phase director in the nematic phase and parallel to and across the layer in the smectic A phase of the OAB smectogen has been measured exploiting a static fringe field gradient 1H NMR diffusometry approach. PMID- 16273141 TI - Investigation of the surface structure of zeolite A. AB - The surface structure of zeolite A was investigated by FE-SEM, AFM, and HRTEM. First, it was shown by FE-SEM and AFM that the surface of zeolite A terminated with the same structure. Then, by combining the results with those of HRTEM, the terminal structure of zeolite A was identified as incomplete sodalite cages. PMID- 16273142 TI - Infrared spectra of the Cl- -C2H4 and Br- -C2H4 anion dimers. AB - Infrared spectra of mass-selected Cl- -C2H4 and Br- -C2H4 complexes are recorded in the vicinity of the ethylene CH stretching vibrations (2700-3300 cm(-1) using vibrational predissociation spectroscopy. Spectra of both complexes exhibit 6 prominent peaks in the CH stretch region. Comparison with calculated frequencies reveal that the 4 higher frequency bands are associated with CH stretching modes of the C2H4 subunit, while the 2 weaker bands are assigned as overtone or combinations bands gaining intensity through interaction with the CH stretches. Ab initio calculations at the MP2/aug-cc-pVDZ level suggest that C2H4 preferentially forms a single linear H-bond with Cl- and Br- although a planar bifurcated configuration lies only slightly higher in energy (by 110 and 16 cm( 1), respectively). One-dimensional potential energy curves describing the in plane intermolecular bending motion are developed which are used to determine the corresponding vibrational energies and wavefunctions. Experimental and theoretical results suggest that in their ground vibrational state the Cl- -C2H4 and Br- -C2H4 complexes are localized in the single H-bonded configuration, but that with the addition of modest amounts of internal energy, the in-plane bending wavefunction also has significant amplitude in the bifurcated structure. PMID- 16273143 TI - Photophysics of ruthenium polypyridyl complexes formed with lacunary polyoxotungstates with iron addenda. AB - The interactions between luminophore [Ru(bpy)3]2+, and the lacunary Dawson heteropolyanions, [P2W17O61(FeOH2)]7-, [P2W17O61(FeBr)]6- and [P2W17O61]10- were investigated using a combination of photophysics, optical and Raman spectroscopy. Extensive quenching of the excited state of [Ru(bpy)3]2+ was observed in each case. Quenching is attributed to the formation of association complexes between [Ru(bpy)(3)]2+ and the heteropolyanions in which the charge on the heteropolyanions is fully compensated for by the ruthenium polypyridyl species. The interaction appears to be principally electrostatic in nature producing [Ru(bpy)3]3.5[P2W17O61(FeOH2)], [Ru(bpy)3]3[P2W17O61(FeBr)] and [Ru(bpy)3]5[P2W17O61]10-. The association constants for formation of the clusters were obtained from photophysical studies and surprisingly, despite the electrostatic nature of the interaction, there was no correlation between the charge on the polyoxometallate and the association constant. In particular, the unsubstituted lacunary, [P2W17O61]10-, showed considerably weaker association compared to the transition metal substituted lacunaries, in spite of its 10- charge. Difference absorption spectroscopy revealed a new transition at ca. 480 nm for each of the cluster complexes. From resonance Raman spectroscopy the origin of this transition was found to involve the polyoxometallate. Unlike previously reported adducts, the cluster complexes formed were not luminescent. In all cases the cluster complexes exhibit remarkable photostability, with no photodecomposition or photo-induced ligand exchange reactions evident in acetonitrile, under conditions where [Ru(bpy)3]2+ alone exhibits considerable photolability. PMID- 16273144 TI - A kinetic model for uptake of HNO3 and HCl on ice in a coated wall flow system. AB - A simple model of gas flow and surface exchange with a single site Langmuir mechanism has been developed to describe effects of adsorption and desorption on trace gas concentrations at the outflow from a coated wall flow tube reactor. The model was tested by simulating experimental results for the uptake of HNO3 and HCl on ice films at temperatures and gas concentrations corresponding to the ice stability region in the upper troposphere. The experimental time-dependent uptake profiles were best fitted with an additional process involving diffusion of the adsorbed molecules into the ice film. The model allowed true surface coverages to be distinguished from total uptake including transfer to the bulk, leading to more accurate estimates of the Langmuir constant, Keq, for surface adsorption. A revised expression was obtained for the temperature dependence of the Keq=-(4.43 +/- 0.77)x 10(5)T+(10.72 +/- 1.75)x 10(7) hPa-1. Reasonable fits to the desorption profiles observed following cessation of exposure of the film to HNO3 or HCl were obtained at high surface coverage but at low coverage desorption was too slow. The analysis suggested that the ice surface was characterised by sites of different binding energy, some weakly bound sites from which the acid molecules desorbed rapidly, and some strong-binding sites which led to essentially irreversible uptake. Experiments involving competitive co-adsorption of HNO3 and HCl, conducted at relatively high equilibrium surface coverage, were well simulated by the model, as were those where the same surface was repeatedly exposed to gas phase acids. PMID- 16273145 TI - Kinetics of model energy landscapes: an approach to complex systems. AB - An idealized potential energy surface (PES), simply a PES-like network of stationary points, is demonstrated to be a useful tool to study kinetic relaxation of complex energy landscapes. Combined with a master equation, we show that if constructed with proper regularity, the kinetics of the PES is easy to predict and understand by carefully examining the eigenmodes of the master equation. By modifying the idealized PES model to make it more and more complicated, we demonstrate a systematic method to study the complex kinetics on large PES. The idealized PES model is used to explore the feasibility and the robustness of statistical sampling of large PES. We develop several sampling strategies, such as the "rough topography sampling" and the "low barrier saddle sampling" in the idealized PES model and find they are also applicable to a realistic PES of the 13-atom Morse cluster with range parameter rho= 6. To measure the robustness of the sampling methods, we compare the eigenvalue spectra, the eigenvector similarity and the relaxation times of the total energy of the full and sample PESs. PMID- 16273146 TI - Time-resolved total internal reflection fluorescence spectroscopy. Part I. Photophysics of Coumarin 343 at liquid/liquid interface. AB - Pico-second time-resolved time-correlated single photon counting (TCSPC) technique under the total internal reflection (TIR) condition has been used to study the photophysical properties of Coumarin 343 (C343) dye molecules adsorbed at the water/1,2-dichloroethane (DCE) interface. The fluorescence decay profile of C343 under TIR condition at the water/DCE interface was non-exponential and fitted to the double exponential decay function with the fluorescence lifetimes 0.3 and 3.6 ns, which proved the existence of two different forms of C343 species having largely different lifetimes at the interface. The longer fluorescence lifetime component of C343 at the interface is attributed to the emission from the monomeric form of the dye molecules and the shorter lifetime component is due to the aggregation of dye molecules. The penetration depth dependence of decay curves indicated no change in the fluorescence lifetime components, however, the amplitude corresponding to the lifetime of aggregate increased and the amplitude corresponding to the lifetime of monomer decreased with the decrease in penetration depth of the aqueous phase from the interface. Aggregation is significant in the interfacial layer. The decrease in monomer lifetime at the interface compared to that in the bulk solution is interpreted in terms of excitation energy migration between the dye molecules. PMID- 16273147 TI - Conformation of glycosaminoglycans by ion mobility mass spectrometry and molecular modelling. AB - We have performed conformational analyses of heparin-derived oligosaccharide ions in the gas phase using a combination of ion-mobility mass spectrometry and molecular modelling. Negative mode electrospray ionisation was used to generate singly (disaccharide, [C12H15NO19S3Na3]-) and doubly charged (tetrasaccharides, [C24H30N2O38S6Na6]2- and [C24H31N2O35S5Na5]2-) ions containing three and six Na+ ions, respectively. Good agreement was obtained between the experimental and theoretical cross sections. The latter were obtained using modelled structures generated by the AMBER-based force field. Analysis of the conformations of the oligosaccharide ions shows that sodium cations play a major role in stabilizing these ions in the gas phase. This was seen in the formation of oligomers of the disaccharide ion and "compact" structures of tetrasaccharide ions. Interestingly, the gas phase conformations of the three tetrasaccharide ions with different primary structures were significantly different. PMID- 16273148 TI - Reduced and oxidized cytochrome c4 exhibit differences in dynamics. AB - The temperature-dependent dynamics of the fully reduced and fully oxidized forms of Pseudomonas stutzeri cytochrome c4 have been studied by Mossbauer spectroscopy. Prior to the dynamic analysis, an efficient labelling strategy has been developed for the expression of highly enriched (57)Fe recombinant cytochrome c4. Subsequently, the protein has been purified to apparent homogeneity. Mossbauer measurements were recorded in the temperature range 77-240 K for both protein forms. A detailed analysis of the high quality spectra is presented. Based on the information obtained from Mossbauer spectroscopy, similarities and differences between cytochrome c4, cytochrome c and HiPIP are discussed. The obtained results reveal that (a) cytochrome c4 exists in pure low spin electronic configuration in both oxidation states in the temperature range 77-240 K, (b) the heme pocket is more relaxed in cytochrome c4 than in cytochrome c, (c) the reduced cytochrome c4 is the most flexible at low temperatures, and (d) protein specific dynamics are most distinct in the oxidized protein. PMID- 16273149 TI - Protein-lipid interactions at the air/water interface. AB - Surface pressure measurements and external reflection FTIR spectroscopy have been used to probe protein-lipid interactions at the air/water interface. Spread monomolecular layers of stearic acid and phosphocholine were prepared and held at different compressed phase states prior to the introduction of protein to the buffered subphase. Contrasting interfacial behaviour of the proteins, albumin and lysozyme, was observed and revealed the role of both electrostatic and hydrophobic interactions in protein adsorption. The rate of adsorption of lysozyme to the air/water interface increased dramatically in the presence of stearic acid, due to strong electrostatic interactions between the negatively charged stearic acid head group and lysozyme, whose net charge at pH 7 is positive. Introduction of albumin to the subphase resulted in solubilisation of the stearic acid via the formation of an albumin-stearic acid complex and subsequent adsorption of albumin. This observation held for both human and bovine serum albumin. Protein adsorption to a PC layer held at low surface pressure revealed adsorption rates similar to adsorption to the bare air/water interface and suggested very little interaction between the protein and the lipid. For PC layers in their compressed phase state some adsorption of protein occurred after long adsorption times. Structural changes of both lysozyme and albumin were observed during adsorption, but these were dramatically reduced in the presence of a lipid layer compared to that of adsorption to the pure air/water interface. PMID- 16273150 TI - Monte Carlo simulation of an inhomogeneous dielectric continuum model for B-DNA. AB - Thermodynamic and structural properties of the counterion atmosphere surrounding B-DNA are calculated by Monte Carlo simulation in a spatially inhomogeneous, but piecewise uniform, dielectric continuum cell model - the "barbarous" model. A boundary element formulation is implemented to study the sensitivity of these properties with respect to perturbations in the location of discontinuous dielectric boundaries relative to fixed and mobile charges. High concentrations are considered corresponding to the liquid crystalline hexagonally ordered phase of DNA. Primitive model results are verified against other simulation reports and a comparison of barbarous model predictions with experimental data is discussed. The internal energy, osmotic coefficient, radial distributions and the population ratio of counterions in the geometrically resolved major and minor grooves are all found to strongly depend on the dielectric boundary position. This suggests that a self-consistent development of the model should consider a free surface problem where the boundary is not specified a priori. PMID- 16273151 TI - Simulation of proliferation of neural stem cells on a surface with emphasis on spatial constraints on cell division. AB - We present Monte Carlo lattice simulations of proliferation of cells on a surface in the situation when the cell-cell adhesion is relatively strong and the cells may form islands and/or flattened hemispheres. The model parameters were chosen to mimic proliferation of adult rat neural stem cells (or, more specifically, adult hippocampal progenitor cells) deposited on polyornithine and laminin coated polystyrene. The results obtained show that the spatial constraints on cell division may result in slowdown of the exponential growth. Depending on the rules used for cell division, this effect may be either nearly negligible or appreciable. In the latter case, the scale of the deviations from the exponential growth is comparable with that observed in our experiments. In the simulations, the slowdown of the growth starts however somewhat earlier and occurs in a less abrupt manner. This seems to indicate that the spatial constraints on division of cells are not the main factor behind the experimentally observed termination of the growth. PMID- 16273152 TI - Guest-host encapsulation of microporous zeolites in ordered mesoporous materials by molecular simulations. AB - The present work provides the first study of ordered mesoporous materials SBA-15 coated with microporous zeolites ZSM-5 using molecular simulations. Several model structures with characteristics such as periodic arrangement of mesopores, randomly arranged micropores, surface hydroxyls and bulk deformations of SBA-15 were used. Cartesian coordinates of ZSM-5 unit lattice were obtained from the literature and the 100 face of H-ZSM-5 unit cell was then placed on the surface of SBA-15 and the entire structure was equilibrated to obtain final configuration. The resulting structure was characterized using simulated small angle and wide angle X-ray diffraction, Connolly surface area (to compare BET area), accessible pore volume for nitrogen molecules (to compare with t-plot volume of micro and mesopores) and methane adsorption at 303 K. The orientation of ZSM-5 on the SBA-15 had no effect on the surface area, pore volume or adsorption capacity. In order to find out if the addition of microporous ZSM-5 should increase the total methane adsorption capacity due to addition of micropores, we studied adsorption on bare and coated SBA-15. However, total adsorption capacity was found to decrease, while the number of methane molecules adsorbed per unit cell of the SBA-15 structure increased. An existing experimental method (J. Am. Chem. Soc., 2004, 126, 14324) of the synthesizing hybrid ZSM-5/SBA-15 structure was studied using accessible micropore volume (by t plot). It was found that the procedure made all the micropores inaccessible. A modification of the method or use of other host materials is suggested to use the benefits of narrow micropore distribution in ZSM-5. PMID- 16273154 TI - Middle aortic syndrome: from presentation to contemporary open surgical and endovascular treatment. AB - Middle aortic syndrome (MAS) is a clinical condition generated by segmental narrowing of the abdominal or distal descending thoracic aorta. MAS may be acquired, caused by Takayasu's or temporal arteritis (giant cell arteritides), neurofibromatosis, fibromuscular dysplasia, retroperitoneal fibrosis, mucopolysaccharidosis, and the Williams syndrome, or congenital, ascribed to a developmental anomaly in the fusion and maturation of the paired embryonic dorsal aortas. Segmental aortic stenosis may be located at the suprarenal, inter-renal or infrarenal aorta, with a high propensity for concomitant stenoses in both the renal (63%) and visceral (33%) arteries. Hypertension proximal to the aortic stenosis, and relative hypotension distal to it, are characteristic findings in MAS. Typical manifestations include headache, early fatigue on exertion, and bilateral lower-limb claudication. The severity of hypertension is the primary indication for intervention and the factor determining procedural timing. As a great proportion of patients with MAS are children or teenagers, the clinical benefits of early surgical intervention to reverse refractory hypertension have to be weighed against the repercussions pertaining to the insult of surgery on the developing aorta. Open surgery is the primary treatment of tubular aortic narrowing (MAS) associated with renovascular hypertension and visceral artery stenosis. This entails aortoaortic bypass of the diseased segment or, less often, patch aortoplasty and usually bypass grafting of the stenosed renal and visceral arteries performed with autologous conduits, particularly in the youngest of patients. Endovascular therapy may provide a sound minimally invasive treatment in MAS caused by discrete aortic stenoses that do not encompass the mesenteric and renal arteries. Hypertension is thus improved or cured in more than 70% of patients. Prognosis after uncompromised surgical reconstruction is rewarding in the mid and long term in patients with congenital aortic coarctation but deteriorates in patients with aortoarteritis and recurrent inflammatory activity. PMID- 16273155 TI - Commentary on "Middle aortic syndrome: from presentation to contemporary open surgical and endovascular treatment.". PMID- 16273157 TI - Thoracoabdominal aortic aneurysm repair: historical review and description of a re-engineered technique. AB - Most complications related to thoracoabdominal aortic reconstruction stem from ischemia-induced injury to the viscera, kidneys, and spinal cord. Pioneers in the treatment of thoracoabdominal aortic aneurysms recognized the danger of producing ischemic damage to these vital organs. In addition to adjunctive methods designed to minimize metabolic demands of the spinal cord during aortic cross-clamping, a variety of extracorporeal techniques have been developed that provide supplemental blood flow to vital end organs during the period of clamp-induced ischemia. This article reviews these extracorporeal methods and provides a historical perspective of thoracoabdominal aortic aneurysm repair. In addition, a reengineered technique for thoracoabdominal aortic aneurysm repair is highlighted. PMID- 16273158 TI - Commentary on "Thoracoabdominal aortic aneurysm repair: historical review and description of a re-engineered technique". PMID- 16273160 TI - Surgical repair of extensive aortic aneurysms. AB - We present our 14-year experience in the management of extensive aortic aneurysms. Significant progress has been made in reducing the morbidity and mortality associated with these procedures. Our strategies for organ protection, operative techniques, including the elephant trunk technique, and surgical results are discussed. PMID- 16273161 TI - Commentary on "Surgical repair of extensive aortic aneurysms". PMID- 16273163 TI - Extra-anatomic visceral revascularization and endovascular stent-grafting for complex thoracoabdominal aortic lesions. AB - Open surgical treatment of lesions of the thoracoabdominal aorta is a formidable procedure. It is associated with significant perioperative morbidity and mortality as well as a prolonged recuperative period. Because of the magnitude of the surgery required, many patients with thoracoabdominal aortic lesions are considered a prohibitive surgical risk and are denied elective surgical intervention. Risk factors for open surgery include prior thoracoabdominal aortic procedures, underlying pulmonary, renal, or cardiac dysfunction, and advanced age. In an effort to reduce the surgical risk, a combined technique has been developed consisting of retrograde extra-anatomic revascularization of visceral vessels followed by thoracoabdominal aortic endografting. Our experience and reports from other institutions suggest that early outcomes are acceptable, even in high-risk patients otherwise deemed unacceptable candidates for open thoracoabdominal surgery. The general principles for patient selection, the surgical and endovascular techniques involved, and representative cases will be presented. PMID- 16273164 TI - Commentary on "Extra-anatomic visceral revascularization and endovascular stent grafting for complex thoracoabdominal aortic lesions". PMID- 16273166 TI - Infrainguinal endovascular procedures should be reserved for patients who do not have good open surgical options. AB - This article is the result of a debate. The motion proposed was "Infrainguinal endovascular procedures should be reserved for patients who do not have good open surgical options.'' Arguments in favor of the motion were offered by Daniel J. Reddy of Henry Ford Hospital in Detroit, MI, and arguments against the motion were offered by Peter Kalman of Loyola University Medical Center in Maywood, IL. PMID- 16273167 TI - Evaluation of macrocirculatory endothelium-dependent and endothelium-independent vasoreactivity in vascular disease. AB - Macrocirculatory endothelium-dependent and independent vasodilatation is integral to tissue-bed oxygen delivery and homeostasis. Dysfunction of macrocirculatory vasoreactivity is a precursor to atherosclerosis and occurs in a similar fashion in multiple tissue beds long before the onset of symptoms. Impaired macrocirculatory vasodilatation has been shown to occur in certain disease states including diabetes mellitus, hypercholesterolemia, chronic renal failure, peripheral arterial atherosclerosis, and abdominal aortic aneurysms, as well as secondary to smoking, advanced age, menopause, high-fat diet, and sedentary lifestyle. Brachial artery vasoreactivity is a noninvasive means of assessing macrocirculatory vasodilatory capacity that may help identify patients at increased risk for peripheral and cardiovascular disease and allow for objective assessment and monitoring of treatment. Endothelium-dependent vasoreactivity, or flow-mediated dilatation, is measured after brachial artery occlusion with a pneumatic blood pressure cuff, and endothelium-independent vasoreactivity is measured after the administration of sublingual nitroglycerin. The accuracy of brachial artery vasoreactivity is dependent on hematologic variables, as well as diurnal, day-to-day, ultrasound operator, and reader variations; however, the overall coefficient of variation is only 1.8%. We discuss the importance of the macrocirculation, investigative methods for evaluating macrocirculatory vasoreactivity, and review the literature of vasoreactivity in these different states. PMID- 16273169 TI - Commentary. Cilostazol inhibits leukocyte integrin Mac-1, leading to a potential reduction in restenosis after coronary stent implantation. PMID- 16273168 TI - Multi-institutional pivotal trial of the Zenith TX2 thoracic aortic stent-graft for treatment of descending thoracic aortic aneurysms: clinical study design. AB - Despite significant improvements in surgical techniques and perioperative management, the repair of descending thoracic aortic aneurysms remains a challenge with the potential for substantial morbidity and mortality. Over the past several years, buoyed by the technical success of endovascular repair of infrarenal abdominal aortic aneurysms, several case series reports of endovascular stent-graft placement for descending thoracic aortic aneurysms have demonstrated the potential safety and efficacy of this treatment modality. Several single-institution studies have documented promising results with these devices, but without controlled clinical trials, the data are insufficient to determine if thoracic aortic endografts provide equivalent or improved outcomes compared with standard open repair. We describe a study design of an ongoing prospective, nonrandomized, multi-institutional, investigational device exemption phase II pivotal clinical trial investigating the safety and effectiveness of the Zenith TX2 thoracic aortic endovascular graft in the elective treatment of patients with descending thoracic aortic aneurysms. PMID- 16273171 TI - Commentary. Weighing the evidence for expanding physician supply. PMID- 16273173 TI - Commentary. Distinct endothelial phenotypes evoked by arterial waveforms derived from atherosclerosis-susceptible and -resistant regions of human vasculature. PMID- 16273176 TI - Commentary. COX-2-derived prostacyclin confers atheroprotection on female mice. PMID- 16273177 TI - Commentary. Meta-analysis: outcomes in patients with suspected pulmonary embolism managed with computed tomographic pulmonary angiography. PMID- 16273179 TI - Commentary. Are patients receiving maximal medical therapy following carotid endarterectomy? PMID- 16273182 TI - Commentary. Early carotid artery stenting and angioplasty in patients with acute ischemic stroke. PMID- 16273183 TI - Commentary. Comparison of surgery and compression with compression alone in chronic venous ulceration (ESCHAR study): randomised controlled trial. PMID- 16273185 TI - Editorial: clear cell sarcoma and osteoclast-rich clear cell sarcoma-like tumor of the gastrointestinal tract: one tumor type or two? Melanoma or sarcoma? PMID- 16273186 TI - Clear cell sarcoma-like tumor with osteoclast-like giant cells in the small bowel: further evidence for a new tumor entity. AB - Most mesenchymal neoplasms of the gastrointestinal tract belong to the category of gastrointestinal stromal tumors (GISTs) and are characterized by the immunohistochemical expression of KIT receptor. In cases without detectable KIT receptor expression several differential diagnoses have to be taken into consideration. Here, we report a case of a 41-year-old man with a tumor of the small bowel composed of large epithelioid tumor cells arranged in solid and alveolar sheets including scattered osteoclast-like multinucleated giant cells. Immunohistochemically, the tumor cells expressed strongly S-100 protein, vimentin, and to a lesser extent, bcl-2. HMB-45, melan-A, KIT receptor, desmin, smooth-muscle actin, and CD-34 were not detectable. Ki-67 index was 20%. The diagnosis was established by 2 different FISH strategies demostrating the presence of a t(12;22)(q13;q12) translocation, the diagnostic hallmark of clear cell sarcoma of soft parts. Our results provide further evidence for the existence of a new tumor entity designated gastrointestinal clear cell sarcoma with osteoclast-like giant cells. The diagnosis of this entity should be considered in the presence of S-100-positive tumors of the gastrointestinal tract containing multinucleated giant cells and can be established by FISH analysis. PMID- 16273187 TI - Remarkably high frequency of EGFR expression in breast carcinomas with squamous differentiation. AB - The human epidermal growth factor receptor (EGFR) is reportedly overexpressed in 15-20% of breast carcinomas. EGFR overexpression is associated with reduced survival and is inversely correlated with expression of estrogen receptor (ER). This study assessed EGFR expression in breast carcinomas with squamous differentiation. The immunohistochemical (IHC) expression of EGFR was evaluated in 39 breast carcinomas with squamous differentiation (30 pure squamous, 6 adenosquamous, 3 carcinosarcomas) by use of the pharmDx assay (clone 2-18C9, DakoCytomation). Cases were considered positive if at least 10% of the cells showed 1+ positivity in the squamous component. Squamous differentiation was confirmed with IHC for CK5-6 (clone D5/16B4, DakoCytomation). ER, PR, and HER2 status as well as clinical information regarding treatment and outcome were correlated. As a control, a tissue microarray comprising 280 lymph node positive breast carcinomas was evaluated with the same EGFR assay. The 39 patients ranged in age from 33 to 77 years (mean 52). The tumors measured 1.3-30 cm (mean 4.8). Sentinel or full axillary lymph node dissection was performed in 28 patients. Fourteen patients had positive lymph nodes. At the time of initial diagnosis, 3 patients had distant metastasis. Follow-up was available for 16 patients (mean 45 months). Disease-free survival at 3 years was 70%. Among the 39 tumors 87% (34) were positive for EGFR (p<0.0001). Sixty-nine percent (27 of 39) showed >50% 2+ EGFR staining. EGFR-positive tumor cells (showing squamous morphology) were also found in 1 bone, 1 lung, and 8 of 11 lymph node metastases available for evaluation. All 11 lymph nodes showed squamous differentiation. All but 1 of the EGFR+ tumors examined were ER and PR negative. Six EGFR-positive tumors were HER2 positive. No statistically significant differences in HER2 status, size, lymph node status and disease-free survival were observed between EGFR+ and EGFR- cases, but the number of EGFR-negative tumors was quite small. Nine of 280 (3%) of lymph node-positive invasive carcinomas on the tissue microarray were EGFR+; review of the initial diagnostic slides failed to reveal squamous features in all but 1 of the 9 EGFR+ tumors. Breast carcinomas with squamous differentiation are a distinct subgroup of breast tumors with a very high frequency of EGFR positivity. Breast carcinomas of this type would be ideal candidates for a trial with EGFR inhibitors. PMID- 16273188 TI - p63 expression in salivary gland tumors: role of DeltaNp73L in neoplastic transformation. AB - To illustrate the role of p63 and its truncated variants in salivary gland tumors, 23 consecutive tumors and 6 normal salivary glands were studied immunohistochemically with anti-p63 antibody and by reverse transcriptase (RT) and nested polymerase chain reaction (PCR) to detect p63 isoform expression. Normal salivary glands: p63 antibody-stained basal and myoepithelial cells; by RT and nested PCR, the 2 main isoforms were present, whereas DeltaNp73L was absent. Tumors: p63 antibody was positive in the following: Warthin tumor (WT) (3/3), oncocytoma (OC) (1/1), pleomorphic adenoma (PA) (7/7), polymorphous-low-grade adenocarcinoma (PLGA) (3/3), adenoid-cystic carcinoma (ADCC)(3/4), epithelial myoepithelial-cell carcinoma (EMC) (1/1), and myoepithelial-cell carcinoma (MCC) (1/1). By RT and nested PCR all tumors expressed p63 irrespective of their morphologic differentiation. The DeltaNp73L isoform was present in tumoral tissue but absent in normal salivary gland. These data suggest that p63, particularly its splice variant DeltaNp73L, is involved in the neoplastic transformation of salivary glands. PMID- 16273189 TI - Biomarker analysis in carcinoma ex pleomorphic adenoma at an early phase of carcinomatous transformation. AB - Diagnostic criteria for intracapsular carcinoma ex pleomorphic adenoma (CXPA) are subjective and vary among authors. Biomarker analysis, which could provide more objective evaluation of these tumors, has rarely been studied in intracapsular CXPA. Immunohistochemical evaluation of c-erbB-2, p53 protein, bcl-2, and Ki-67 was performed in 8 cases of CXPA at an early phase of malignant transformation (4 intracapsular and 4 minimally invasive) and in 17 pleomorphic adenomas (PA). In all cases of CXPA, p53 and Ki-67 were demonstrated predominantly in luminal cells of benign and malignant areas, significantly more in the latter. Few benign myoepithelial cells were p53 positive. c-erbB-2 reactivity was strongly associated with atypical luminal cells. Bcl-2 expression was weak and focal in malignant areas from 2 cases. In conclusion, both p53 and c-erbB-2 proteins appear to be involved at an early stage of malignization of PA. In PA with atypical cells, evaluation of the expression of these 2 markers provides more objective criteria for the diagnosis of intracapsular CXPA. PMID- 16273190 TI - Role of chromogenic in situ hybridization (CISH) in the evaluation of HER2 status in breast carcinoma: comparison with immunohistochemistry and FISH. AB - We report our experience with Chromogenic in Situ Hybridization (CISH) for the evaluation of HER2 amplification on 55 cases of formalin-fixed, paraffin-embedded invasive breast carcinomas of different histology. All the results were corrected for chromosome 17 aneusomy and compared with immunohistochemistry (IHC); a subset of cases was compared to FISH. Thirty-one of 32 cases in which FISH and CISH were performed yielded the same results. CISH and IHC showed a good concordance in the 0/1+ and 3+ category, while a poor agreement with weakly protein overexpression was confirmed. Chromosome 17 analysis was necessary in cases with a low number of HER2 gene copies. CISH is a useful tool to evaluate breast cancer HER2 status that can be easily implemented in a laboratory of surgical pathology. PMID- 16273191 TI - Antigen retrieval methods and estrogen receptor immunoexpression using 1D5 antibody: a comparative study. AB - Long-term formalin tissue fixation results in antigen masking, probably through aldehydic linkage between proteins and fixative molecules. Immunohistochemistry results depend on the type of the detection procedure and the type of antibody used for the reaction. Considering the difficulty in working with estrogen receptor (ER) antibodies and the lack of standardization of the antigen retrieval methods, we quantified the immunoexpression of ER using the 1D5 antibody and a standard streptavidin-biotin detection procedure retrieving with microwave oven, steamer, pressure cooker, and water bath in a set of SBR grade 2 invasive breast carcinomas. Pressure-cooking provided the best results. No significant differences were observed in using the other methods. Pressure-cooking should be recommended as the method of choice for standardization of the ER immunohistochemical reaction. PMID- 16273192 TI - Images in pathology: Snoopy in flight gear. PMID- 16273194 TI - Images in pathology: lymphocyte rosettes in an aspirate of bone marrow involved by T-Cell-Rich B-Cell lymphoma (TCRBCL). PMID- 16273193 TI - Images in pathology: granule cell loss in AIDS-associated progressive multifocal leukoencephalopathy. PMID- 16273195 TI - Images in pathology: ectopic embryo in a glomerulus. PMID- 16273196 TI - Images in pathology: parotid bunny eating carrot. PMID- 16273197 TI - Osteoblastoma of the larynx: a case report. AB - We describe a unique case of an osteoblastoma in the cricoid cartilage of a 50 year-old man who presented with hoarseness and progressive dyspnea. Panendoscopic examination revealed a reddish tumor at the left side of the cricoid cartilage. Computed tomography (CT) imaging showed a sclerotic lesion involving the cricoid cartilage, protruding in the airway lumen. The tumor was removed via an external procedure. Histology confirmed the diagnosis of osteoblastoma. PMID- 16273198 TI - Anaplastic large cell lymphoma: a unique presentation with urinary bladder involvement: a case report. AB - Anaplastic large cell lymphoma (ALCL) is a T-cell lymphoma composed of large pleomorphic CD30-positive cells. While systemic ALCL frequently involves extranodal sites, involvement of the urinary bladder is extremely rare. We report a case of systemic ALCL presenting with bladder involvement. A 28-year-old man presented with hematuria, dysuria, and lower abdominal pain. Imaging revealed pelvic lymphadenopathy and a thickened bladder wall. Bladder biopsies showed diffuse infiltration of the lamina propria by large pleomorphic cells, with preservation of the overlying urothelium. Immunohistochemistry demonstrated cell membrane and Golgi region staining for CD30 and epithelial membrane antigen. This is the first documented instance of systemic ALCL presenting with bladder symptoms. PMID- 16273199 TI - Urothelial signet-ring cell carcinoma of the renal pelvis with collagenous spherulosis: a case report. AB - We present a unique case of urothelial carcinoma of the right renal pelvis. It occurred in a 58-year-old woman. The tumor was located in the renal pelvis with extension into the adjacent renal medulla and cortex. Two years after surgical excision the patient is free of recurrence and metastasis. The tumor was well demarcated, without capsule, firm, solid, and whitish on the cut surface. It was 3x4 cm in largest diameter and without signs of necroses and hemorrhages. The tumor did not infiltrate the ureter. Histologically the predominant pattern of the tumor was adenocarcinomatous differentiation, and only very rare foci of urothelial carcinoma composed of typical transitional cells were found. No signs of intestinal type of metaplasia and adenocarcinoma, changes similar to the cystitis cystica or cystitis glandularis, were found in the tumor or in its vicinity. Most of the tumor looked like solid nests composed of cells with intracytoplasmic lumens. The resulting appearance was that of typical signet-ring cell change. These solid nests were usually surrounded by columnar epithelium, which in many areas formed papillary structures. A very striking feature was formation of collagen spherules. Small collagen spherules were often surrounded by a layer of the neoplastic cells so that collagenous rosettes were formed. In some areas these collagenous spherules clustered together so that they formed areas of collagenous spherulosis. The collagen in the spherules reacted positively with collagen IV. Ultrastructurally these spherules were formed by basal membrane-like material. Intracytoplasmic lumens of the signet-ring cell change were endowed by slender microvilli at ultrastructural level. PMID- 16273200 TI - Peribiliary cysts can mimic Caroli's disease: a case report. AB - Peribiliary cysts, otherwise known as cystic dilatation of the peribiliary glands, are uncommon, and are usually discovered incidentally at autopsy, or in explants following liver transplantation. Preoperative diagnosis is often difficult owing to their asymptomatic nature and small size. Exclusion of a premalignant or malignant cystic condition is mandatory. We report a case of peribiliary cysts, initially thought to represent Caroli's disease, and briefly discuss the management of this condition. PMID- 16273202 TI - Identification of SP5 as a downstream gene of the beta-catenin/Tcf pathway and its enhanced expression in human colon cancer. AB - Mutations in APC, CTNNB1, AXIN1 or AXIN2 cause impairment in the beta-catenin degradation pathway and result in accumulation of beta-catenin in a wide range of human cancers. Accumulated beta-catenin then associates with Tcf/LEF transcription factors and transactivates their target genes. To uncover in detail the role of accumulated beta-catenin in colorectal carcinogenesis, we searched for genes involved in the beta-catenin/Tcf signaling pathway by cDNA microarray. We identified and characterized a human gene, SP5, that was down-regulated after depletion of beta-catenin by transduction of wild-type APC into SW480 cells. SP5 is a member of the Sp transcription factor family, which binds to the GC box or closely related sequences in promoters of many genes and control their expression. Reporter assays and an electromobility-shift assay revealed a DNA fragment between -285 and -279 in the 5' flanking region of this gene to be a target of the beta-catenin/Tcf4 complex. Our results indicate that SP5 is a novel direct down-stream target in the Wnt signaling pathway. PMID- 16273201 TI - Cluster analysis of S100 gene expression and genes correlating to psoriasin (S100A7) expression at different stages of breast cancer development. AB - Gene expression patterns in ductal carcinoma in situ (DCIS) and invasive and metastatic breast tumors have been determined using serial analysis of gene expression (SAGE). The purpose of this approach was to identify biologically and clinically meaningful subgroups of DCIS with a high risk of progression to invasive disease. The analyses have led to the identification of several differentially expressed genes, such as HIN-1, dermcidin and S100A7 (psoriasin). The aim of the present study was further to delineate the expression profile of S100 genes using information from 22 breast epithelial SAGE libraries. We demonstrated the down-regulation of S100A6 and S100A10 in breast cancer, irrespective of pathological stage. S100P and S100Z were both up-regulated in cancer; whereas S100A7, S100A8 and S100A9 were strongly up-regulated only in DCIS. The hierarchical clustering of S100 gene expression in these 22 libraries revealed two major groups with distinguishable S100 gene expression profiles. One of them was characterized by the high concomitant expression of S100A7, S100A8 and S100A9. Using SAGE informatics, we found 21 genes with a high positive correlation to S100A7 expression in libraries representing different categories of tissues archived at SAGE Genie, suggesting a function of psoriasin that is not tissue specific. Like S100A7, several of these genes displayed cation-binding properties. We also report the strong correlation in the breast epithelial SAGE libraries between the expression of S100A7 and genes reported as being up regulated in DCIS, as well as in the inflammatory skin disorder, psoriasis; including RGS5, UPK1A, TMPRSS3, S100A9, p53, SCCA1, SCCA2 and KRT17. PMID- 16273203 TI - Vesnarinone inhibits angiogenesis and tumorigenicity of human oral squamous cell carcinoma cells by suppressing the expression of vascular endothelial growth factor and interleukin-8. AB - Vesnarinone is a synthesized positive oral inotropic agent that has multiple biological activities on mammalian cells both in vitro and in vivo. This agent has been reported in relation to its antitumor effect with apoptosis-inducing activity. In the present study, we determined whether vesnarinone could suppress angiogenesis and growth of human oral squamous cell carcinoma cells in vitro and in vivo. Vesnarinone significantly inhibited the in vitro and in vivo expression of two major proangiogenic molecules, vascular endothelial growth factor (VEGF) and interleukin-8 (IL-8), in cultured cells and in cells implanted into the subcutaneous tissue of nude mice. Also, vesnarinone inhibited the nuclear factor kappa B (NF-kappaB) activity in human oral squamous cell carcinoma cells in vitro. The decreased expression of VEGF and IL-8 correlated with decreased tumorigenicity and decreased vascularization of lesions in vivo. These findings suggest that vesnarinone can suppress the angiogenesis and growth of oral squamous cell carcinoma cells by inhibiting the expression of VEGF and IL-8 involved in blockade of NF-kappaB activity. PMID- 16273204 TI - A mutation in the common docking domain of ERK2 in a human cancer cell line, which was associated with its constitutive phosphorylation. AB - The EGFR/Ras/Raf/MEK/ERK pathway is a major pathway involved in the control of growth signals, cell survival and differentiation. Mutations of signaling components, such as EGFR (c-erbB1), Ras, and B-Raf, have been shown to play roles in the genesis of human cancer, while point mutation of ERK has not been reported. In this study, we present evidence for a mutation in an oral squamous cell carcinoma cell line, HSC6. PCR-amplification of cDNA, cloning and sequencing resulted in the identification of glutamic acid to lysine substitution at codon 322 (E322K) that occurred in the common docking (CD) domain of ERK2. The mutant protein contributed towards faster-migration in SDS-PAGE, and constitutive phosphorylation in a MEK-dependent manner. The transient transfection of the mutant ERK2 in 293T cells resulted in the expression of the same faster-migrating band in SDS-PAGE as was detected in HSC6 cells, which was preferentially phosphorylated relative to endogenous wild-type ERK2. The present study is the first to report ERK2 substitution mutation in a human cancer cell line which resulted in constitutive phosphorylation. PMID- 16273206 TI - Expression of the licensing factors, Cdt1 and Geminin, in human colon cancer. AB - Licensing of chromatin for replication is an evolu-tionarily conserved step in the control of cell division and genomic integrity. Proteins that participate in licensing have been recently documented to denote the proliferative state of cells and they have been proposed as diagnostic and prognostic markers in human cancer. Cdt1 was recently discovered as an important licensing factor, that is inhibited by Geminin. In the present study we analyzed Cdt1 and Geminin expression in human colon cancer. We showed that Cdt1 protein is highly expressed in human neoplastic lesions of the colon while its cell-cycle phase-specific expression profile appears preserved during human carcinogenesis. Similarly, Geminin, Cdt1's inhibitor, is also overexpressed in colon carcinomas and its expression correlates with significant clinicopathological parameters of the disease. Moreover, both Cdt1 and Geminin expression are severely downregulated upon differentiation of Caco-2 cells, an in vitro model of intestinal epithelial differentiation. PMID- 16273205 TI - Leptin and leptin receptor expression in the myometrium and uterine myomas: Is leptin involved in tumor development? AB - Leptin, the product of the obesity (ob) gene, along with its receptors (Ob-Rs), is expressed in several tissues and organs. Evidence has been provided that leptin, in addition to being involved in obesity development, plays a role in the regulation of the female reproductive system, angiogenesis and tumor growth. Uterine myoma is a rather common disease that develops more frequently in obese than lean women, where plasma leptin concentrations are elevated. RT-PCR and Western blotting showed that leptin was expressed, as mRNA and protein, in several uterine myomas but not in normal myometrium, while leptin receptors were expressed in both tissues. Immunocytochemistry indicated that leptin immunoreactivity was located in both myometrial cells and blood-vessel walls of uterine myomas. Leptin(22-56), at concentrations of 10(-7) and 10(-6) M, enhanced the proliferative activity of both the normal myometrium and myoma cells in primary culture. Taken together, our findings allow us to suggest that leptin, acting through autocrine-paracrine mechanism(s), may be involved in the development of uterine myomas. PMID- 16273207 TI - Synchronous endometrial and ovarian carcinomas: analysis of genetic relationship of the tumors. AB - Simultaneous carcinomas of the endometrium and ovary may represent a diagnostic dilemma and the clinical management of such cases may be problematic. In surgical pathology practice, we classify them either as double primary tumors (DP) or a single primary tumor with metastasis (PM), according to the conventional clinicopathologic criteria. The distinction has important therapeutic and prognostic implications, but it can be sometimes difficult, especially in advanced cases. In this study, in addition to the clinicopatho-logic classification, we assessed tumor cell clonality in 13 cases with synchronous endometrial and ovarian endometrioid adenocarcinomas using PCR-based microsatellite analysis of microdissected archival tissues for loss of heterozygosity (LOH) and microsatellite instability (MSI). All paired endometrial and ovarian tumors demonstrated either MSI or/and LOH except for 1 case, and therefore the microsatellite analysis was informative in 92.3% of the cases. Nine of 26 tumors (34.6%) exhibited MSI-H and 15 of 26 (57.7%) showed LOH. In contrast to 4 DP cases and 9 PM cases classified according to clinicopathologic criteria, microsatellite analysis suggested 10 DP cases and 2 PM cases. The molecular analysis was not informative in 1 case. Thus, analysis of microsatellite abnormality is a helpful adjunct in the assessment of synchronous tumors, especially to differentiate DP from PM cases in advanced tumor cases. Moreover, the combination of conventional clinicopathologic evaluation and molecular analysis is important and helpful in distinction between the two groups of synchronous tumors. PMID- 16273208 TI - Vitamins K1 and K2 potentiate hyperthermia by down-regulating Hsp72 expression in vitro and in vivo. AB - Hyperthermia is used to treat various malignancies, including esophageal, stomach and rectal cancer. Since hyperthermia alone has produced limited results, much attention has been focused on combining hyperthermia with chemotherapy and on searching for substances able to sensitize tumor cells to hyperthermia-induced damage. Here, we show that vitamins K1 and K2 (VK1, VK2) inhibited the expression of heat-shock protein 72 (Hsp72) but did not affect the constitutive expression of Hsc70 or calnexin in vitro and in vivo. VK1 and VK2 sensitized A549 cells to heat-shock induced cell death, while the compounds alone had no effect on cell viability. The suppression of Hsp72 was apparently at the protein level because the mRNA expression of Hsp72 was unchanged. Moreover, the chaperone activity of Hsp72 was compromised after heat-shock when cells were pre-treated with VK2. The effect of VK2 on Hsp72 suppression, however, was also observed in normal mouse tissue after the mice were subjected to whole-body hyperthermia. To eliminate this side effect, local hyperthermia was performed on tumors in mice. The pre treatment with VK2 potentiated the effect of local hyperthermia on tumor growth suppression. The findings here that VK1 and VK2 inhibit heat-shock-induced Hsp72 suggest their possible use as an adjuvant for hyperthermia in cancer therapy. PMID- 16273209 TI - Expression of SIP1 in oral squamous cell carcinomas: implications for E-cadherin expression and tumor progression. AB - Loss of E-cadherin expression allows carcinoma cells to liberate from the primary site and enhances invasion and metastasis. The genetic aberration of E-cadherin is a rare event in sporadic carcinomas, and transcription repressors are considered to take a central role in E-cadherin loss. However, expression of E cadherin repressors is largely dependent on tissue and cell type. To identify the repressor expressed in oral squamous carcinomas, we compared the expression levels of E-cadherin and repressors by real-time RT-PCR. Among the repressors including SNAIL, SLUG, SIP1, E12 and E47, SIP1 was inversely correlated to E cadherin (P < 0.05). Chromatin immunoprecipitation showed that SIP1 specifically bound to the E-cadherin promoter region. SIP1 expression was immuno histochemically detected in 27.7% of 47 oral carcinomas, and SIP1-positive carcinomas did not express E-cadherin (P < 0.01). Thirteen patients with SIP1 staining showed a lower disease-specific survival rate (P < 0.05). Multivariate risk factor analysis demonstrated that SIP1 expression was an independent prognostic value for disease-specific overall survival (P < 0.05). These results suggest that SIP1 contributes to the loss of E-cadherin expression and that detection of SIP1 expression is a predictive and prognostic tool in clinical management of oral carcinomas. PMID- 16273210 TI - Hypericin: a promising fluorescence marker for differentiating between glioblastoma and neurons in vitro. AB - The naturally occurring photosensitizer, hypericin, with its high quantum yield of singlet oxygen photogeneration was studied for its ability to differentiate between glioblastoma cells and fetal rat neurons using fluorescence microscopy. Eight human glioma cell lines and twelve primary human glioma cell cultures were compared to human astrocytes and cerebellar granule neurons after incubation with 20 microM hypericin for 5-120 min. Photobleaching effects have been studied by exposing the cell lines to 100 msec of excitation light (510-550 wavelength). Mainly, perinuclear hypericin staining was detected. Neurons can be differentiated from glioblastoma cell lines and astrocytes by a lower fluorescence intensity (Tukey-Kramer HSD test, p < 0.0001). Therefore, hypericin seems to be a promising substance for the photodynamic therapy of malignant brain tumors. PMID- 16273211 TI - Dehydroepiandrosterone causes hyperplasia and impairs regeneration in rat liver. AB - DHEA, a steroid of the adrenal gland, is a non-genotoxic hepatocarcinogen of the peroxisome proliferator type in rodents. However, DHEA also exerts anti carcinogenic effects by reducing the number and proliferation of preneoplastic and neoplastic lesions induced by N-nitrosomorpholine. The mechanism underlying this growth-modulating effect is unclear, and no data are available on DHEA effects on normal liver. Here, we show that DHEA is a liver mitogen, increasing proliferation in rat liver after 3 days of treatment (100 mg/kg body weight per day) as indicated by significantly enhanced expression of cyclin E, PCNA and Ki 67 and an elevated number of mitotic figures. Histological observation of the liver and biochemical evaluation of serum transaminases and bilirubin did not reveal any evidence for cell death, demonstrating that increased proliferation was not due to liver damage. After 2 weeks of DHEA-treatment, proliferation parameters returned to control values and, after 4 weeks, cyclin E and Ki-67 were even lower than in controls. To study the DHEA effect on regenerating liver, we performed partial hepatectomy (PHx) on rats pretreated for 4 weeks with DHEA and analyzed the kinetics of the cell cycle. DHEA-treatment delayed the entry of hepatocytes into G1 phase by about 6 h indicated by a later rise in Ki-67 and cyclin E expression. Reduced STAT-3 activation before G1-phase entry indicates an impaired recruitment of hepatocytes to regenerative proliferation in DHEA-treated livers. The rise in proliferation observed after PHx in DHEA-treated livers was more flat and, in contrast to controls, did not show a peak value within the first 35h as indicated by Ki-67, PCNA, cyclin E and BrdU-incorporation levels in hepatocytes. In conclusion, the results show that DHEA acts as a mitogen in rat liver but reduces the regenerative capacity of the liver. PMID- 16273212 TI - Dielectrophoresis-based 'Lab-on-a-chip' devices for programmable binding of microspheres to target cells. AB - There is a general agreement on the fact that the Laboratory on chip (Lab-on-a chip) technology will enable laboratory testing to move from laboratories employing complex equipments into non-laboratory settings. In this respect, dielectrophoresis (DEP) is a very valuable approach to design and produce Lab-on a-chip devices able to manipulate microparticles and cells. In this study, we report the application of DEP-based devices for facilitating programmable interactions between microspheres and target tumor cells. We used two Lab-on-a chip devices, one (the SmartSlide) carrying 193 parallel electrodes and generating up to 50 cylinder-shaped DEP cages, the other (the DEP array) carrying 102,400 arrayed electrodes and generating more than 10,000 spherical DEP cages. We determined whether these devices can be used to levitate and move microspheres and cells in order to obtain a forced interaction between microspheres and target cells. The first major point of this manuscript is that the DEP-based SmartSlide can be used for transfection experiments in which microspheres and target cells are forced to share the same DEP cage, leading to efficient binding of the microspheres to target cells. The data obtained using the DEP array show that this system allows the sequential, software-controlled binding of individually and independently moved single microspheres to a single target tumor cell. To our knowledge, this is the first report on the possible use of a DEP-based Lab-on-a chip device for guided multiple binding of singularly moved microspheres to a single tumor cell. This approach can be of interest in the field of drug discovery, delivery and diagnosis. PMID- 16273213 TI - A model for PTCH1/Ptch1-associated tumors comprising mutational inactivation and gene silencing. AB - Mutations of the Sonic hedgehog (SHH) receptor, Patched1 (PTCH1), have been identified in a variety of tumors. PTCH1 is usually considered to be a tumor suppressor gene. However, one normal allele is retained in many tumors. We investigated the mechanism of tumorigenesis in murine heterozygous Ptch1 knock out mice. Here we show that Ptch1 transcripts, which are consistently overexpressed in tumors in these mice, are derived predominantly from the mutated allele. These transcripts give rise to a mutant protein incapable of pathway inhibition. In contrast, the expression of wild-type transcripts in the tumor is reduced. The transcriptional activity of a Ptch1 promoter is sensitive to methylation. Based on these results, we propose a model, in which tumorigenesis begins with the transcriptional silencing of one PTCH1/Ptch1 allele. This alone has no functional consequences. Upon mutational inactivation of the other allele, the resulting loss of PTCH1/Ptch1 function activates PTCH1/Ptch1 transcription from the non-silenced, i.e. the mutant, allele. These events can occur in an opposite order. This model is consistent with the expression of PTCH1/Ptch1 derived transcripts and proteins found in tumors, with the sensitivity of the murine Ptch1 promoter to methylation, and with the recently reported effect of demethylating agents on Ptch1 expression. These latter agents could be effective in treatment of, at least, some tumors associated with loss of PTCH1 function. PMID- 16273214 TI - Evaluation of p16 and Id1 status and endogenous reference genes in human chondrosarcoma by real-time PCR. AB - Both the tumour suppressor, p16, and the helix-loop-helix transcription factor, Id1, have been assigned roles in tumour growth in general and appear to be involved in chondrosarcoma. Id1 has further been found to repress the expression of p16. Therefore, the mRNA expression of these two genes was studied by real time PCR in a search for prognostic markers in human chondrosarcoma. To get reliable quantitative data, however, the choice of endogenous reference gene for use in the assay is important. Therefore, eleven different endogenous reference genes were evaluated in chondrosarcoma cells and articular chondrocytes. 18S rRNA appeared to be the best choice to use as endogenous reference gene, since it was suitable for both kinds of cells. Several of the other reference genes tested showed variation between individuals or between normal chondrocytes and chondrosarcoma cells. This demonstrates the importance of using a correct endogenous reference gene to get reliable results from quantitative measurements. Both p16 and Id1 showed varied gene expression patterns among the samples and none of these genes could be significantly correlated to prognosis. PMID- 16273215 TI - Telomeres, telomerase and oral cancer (Review). AB - Oral squamous cell carcinoma (oral cancer) and many squamous cell carcinomas of the head and neck arise as a consequence of multiple molecular events induced by the effects of various carcinogens related to tobacco use, environmental factors, and viruses in some instances (e.g., mucosal oncogenic human papillomaviruses), against a background of inheritable resistance or susceptibility. Consequent genetic damage affects many chromosomes and genes, and it is the accumulation of these changes that appears to lead to carcinoma. Telomere maintenance by telomerase or, in its absence, alternative lengthening of telomeres protect this acquired altered genetic information ensuring immortality without losing eukaryotic linear DNA; when this does not occur DNA is lost and end-replication problems arise. Telomerase is reactivated in 80-90% of cancers thus attracting the attention of pathologists and clinicians who have explored its use as a target for anticancer therapy and to develop better diagnostic and prognostic markers. In the last few years, valuable research from various laboratories has provided major insights into telomerase and telomeres leading to their use as diagnostic and prognostic markers in several types of cancer. Moreover, many strategies have emerged which inhibit this complex enzyme for anticancer therapy and are one step ahead of clinical trials. This review explains the basic biology and the clinical implications of telomerase-based diagnosis and prognosis, the prospects for its use in anticancer therapy, and the limitations it presents in the context of oral cancer. PMID- 16273216 TI - Voacamine, an alkaloid extracted from Peschiera fuchsiaefolia, inhibits P glycoprotein action in multidrug-resistant tumor cells. AB - Multidrug resistance (MDR) in tumor cells is generally associated with increased efflux of the cytotoxic compounds, due to the activation of mechanisms of intracellular transport and to the overexpression of surface proteins, such as P glycoprotein (Pgp), which act as ATP-dependent molecular pumps. In a previous study, voacamine, a bisindolic alkaloid from Peschiera fuchsiaefolia, was examined for its possible capability of enhancing the cytotoxic effect of doxorubicin (DOX) on resistant human osteosarcoma cells. The effects of voacamine on the cell survival and on accumulation of DOX were investigated on both the parental cell line, U-2 OS-WT, and its resistant counterpart, U-2 OS-R. A differential effect between sensitive and resistant cells on the intracellular DOX concentration and distribution was revealed. In particular, voacamine induced a significant increase of drug retention and intranuclear location in resistant cells. Moreover, the cell survival analysis and the electron microscopic observations revealed an enhancement of the cytotoxic effect of DOX induced by the plant extract. In the present study, a panel of monoclonal antibodies (MAbs), recognizing different and specific structural and functional state of Pgp, was used. By flow cytometry and immunofluorescence confocal microscopy, a dose dependent increase of the reactivity of Pgp with MAb UIC2, which specifically recognizes an epitope of the drug transporter in its functional conformation, was detected in voacamine-treated U-2 OS-R cells. Conversely, the expression of the epitope recognized by MAb MC57 was downregulated while MAb MM4.17 did not change its binding level to treated and untreated MDR cells. These data suggest that the plant extract reacts with Pgp producing conformational changes with consequent epitope modulation. Taken together, our observations seem to demonstrate that voacamine is a substrate for Pgp and, therefore, interferes with the Pgp-mediated drug export, acting as a competitive antagonist of cytotoxic agents. PMID- 16273217 TI - The molecular mechanisms responsible for resistance to ET-743 (Trabectidin; Yondelis) in the Ewing's sarcoma cell line, TC-71. AB - Identification of new active agents against sarcoma is considered an important challenge in medical oncology. ET-743 (Trabectidin; Yondelis) has recently emerged as the first active drug developed against sarcoma in the last two decades, with promising results especially against soft-tissue sarcoma and Ewing's sarcoma (ES). In this study, we analyzed the molecular mechanisms responsible for resistance to ET-743 in ES cells. Three resistant cell variants (TC/ET 3 nM, TC/ET 6 nM and TC/ET 12 nM) were obtained, showing 28-, 47- and 102 fold increase in ET-743 resistance. Cross-resistance to other drugs was analyzed. Comparative genomic hybridization and cDNA microarray technology were employed to characterize and compare the gene expression profile of two TC/ET variants with the parental cell line. TC/ET cells show a conventional multidrug resistance phenotype and P-glycoprotein overexpression was found to significantly contribute to ET-743 resistance. However, functional studies with the cyclosporine analogue, PSC-833, indicate that other mechanisms are involved in resistance to ET-743. The gene expression profile of TC/ET cells indicated, among up-regulated genes, an increase in expression of insulin-like growth factor receptor-I (IGF-IR) and one of its major intracellular mediators, insulin receptor substrate-1. Functional studies using a neutralizing antibody anti-IGF-IR confirmed involvement of this signaling pathway in resistance to ET-743. Simultaneous blockage of both P glycoprotein and IGF-IR completely restored sensitivity to ET-743 in ES cells. Overall, these findings provide impetus for future studies testing the therapeutic value of new specific inhibitors of P-glycoprotein and IGF-IR, which could represent a concrete therapeutic option for ES patients refractory to conventional agents. PMID- 16273218 TI - Doxazosin induces apoptosis in LNCaP prostate cancer cell line through DNA binding and DNA-dependent protein kinase down-regulation. AB - Doxazosin is a quinazoline-based compound acting as an alpha-1-adrenergic inhibitor shown to induce apoptosis in prostate cancer cell lines via an alpha-1 adrenergic receptor-independent mechanism. To better understand the mechanism of doxazosin-induced apoptosis in prostate cancer, we performed cDNA microarray to analyze gene expression changes produced by doxazosin in the androgen-dependent human prostate cancer cell line, LNCaP. We found that 70 and 92 genes were deregulated after 8 and 24 h of doxazosin treatment, respectively. These genes are involved in several cellular processes such as cell-cycle regulation, cell adhesion and signal transduction pathways. Strikingly, we found that doxazosin induces deregulation of genes implicated in DNA replication and repair, such as GADD45A, XRCC5 and PRKDC. These facts, together with the demonstration of the ability of doxazosin to bind DNA, allowed us to propose a novel mechanism of action for doxazosin in prostate cancer cells that implies DNA-damage mediated apoptosis by down-regulation of XRCC5 and PRKDC genes. PMID- 16273219 TI - An autocrine loop directed by the vascular endothelial growth factor promotes invasiveness of human melanoma cells. AB - The vascular endothelial growth factor-A (VEGF-A) is a cytokine that promotes angiogenesis through the activation of two tyrosine kinase receptors, VEGFR-1 and VEGFR-2, on vascular endothelial cells. Moreover, several experimental evidences indicate that VEGF-A may also play a role in tumor progression by acting on neoplastic cells expressing VEGFRs. In this study we show that human melanoma cells that simultaneously produce VEGF-A and express VEGFRs exhibit a higher spontaneous ability to invade the extracellular matrix (ECM) than melanoma cells not expressing either VEGF-A or VEGFRs. Exposure of VEGFR expressing melanoma cells to exogenous VEGF-A further increases their ability to invade the ECM. Moreover, an inhibitor of VEGFR tyrosine kinase activity is able to abrogate VEGF A-induced stimulation of ECM invasion. A cell clone (13443/N2) derived from a VEGF-A responsive melanoma cell line and expressing high levels of VEGFR-2 invades the ECM eight-fold more efficiently than a cell clone derived from the same cell line and expressing extremely low levels of the receptor. Exposure of 13443/N2 cells to VEGF-E, which selectively binds and activates VEGFR-2, increases their ability to invade the ECM. Finally, the expression of the VEGF-A mRNA antisense sequence in 13443/N2 cells markedly reduces the release of VEGF-A and ECM invasion. In conclusion, our data show for the first time that a VEGF-A driven autocrine loop promotes human melanoma cell ability to invade the ECM, and strongly support the hypothesis that activation of VEGFR-2 plays a primary role in this process. PMID- 16273220 TI - The Liverpool Lung Project research protocol. AB - The Liverpool Lung Project (LLP) is being conducted within a defined geographic area of Merseyside, based on contiguous districts with a high incidence of lung cancer. As well as increasing the scientific understanding of the interactions between the different risk factors for lung cancer, the LLP aims to develop a model which predicts lung cancer risk from an extensive record of epidemiological risk factors, molecular/genetic risk factors and from changes in molecular biomarkers, which maybe used in early detection programmes. The LLP has two components: Firstly, a case-control study of 800 newly diagnosed cases of lung cancer matched 2:1 for age (+/-2 years) and gender will facilitate the development of the molecular genetic and epidemiological model to assess an individual's risk of developing lung cancer. Secondly, a prospective cohort study will recruit 7,500 individuals to identify markers of pre-clinical carcinogenesis in a high-risk population and validate and strengthen the molecular genetic and epidemiological risk assessment model. Structured questionnaires collect detailed information on lifestyle factors (active and passive smoking, medical history, family cancer history, occupation, residence and diet) over the whole life course. Collection of biological specimens includes blood, sputum, bronchial lavage, oral scrapes and tumour. Cohort study subjects will be followed up over a 10-year period. In addition, serial biological samples (blood and induced sputum) will be collected from a high-risk subgroup of the cohort. The LLP protocol provides detailed information on the study design and analysis strategy, together with the informed consent documentation and the LLP lifestyle questionnaire. Conclusions from the LLP programme will be based around epidemiological and genotyping risk assessment models enabling us to identify high-risk populations, and will ultimately contribute to, and facilitate the development of effective prevention, early detection and chemoprevention strategies. PMID- 16273221 TI - Complex agonist-like properties of ICI 182,780 (Faslodex) in human breast cancer cells that predominantly express progesterone receptor-B: implications for treatment resistance. AB - ICI 182,780 (Faslodex), considered a pure anti-estrogen, is approved for treatment of post-menopausal breast cancer patients who fail to respond to tamoxifen therapy. We recently reported that, like mifepristone, ICI 182,780 exhibits anti-progestin activity, blocking the progestin-dependent increase in endogenous vascular endothelial growth factor (VEGF) mRNA and protein release. Some anti-progestins have partial agonist-like activity in breast cancer cells expressing high levels of progesterone receptor B (PRB). Our results show that ICI 182,780 can also induce reporter activity from a plasmid containing a simple progestin responsive element (PRE) in these cells. Using small interfering RNA, we determined that induction is dependent on the presence of PR, estrogen receptor and SRC-1. Regulation of more complex progestin-responsive promoters was context-dependent; induction was observed from the MMTV promoter but not from the VEGF promoter. In contrast, ICI 182,780 increased the release of angiogenically active VEGF from cells expressing elevated levels of PRB. This effect was dependent on the phosphatidylinositol-3 kinase and ERK/MAPK signaling pathways. We hypothesize that these agonist-like properties of ICI 182,780 (one genomic and one non-genomic) may contribute to the acquisition of drug resistance, suggesting that both anti-hormonal and anti-angiogenic treatment may be appropriate in these patients. PMID- 16273222 TI - Expression of p63, COX-2, EGFR and beta-catenin in smokers and patients with squamous cell carcinoma of the head and neck reveal variations in non-neoplastic tissue and no obvious changes in smokers. AB - Squamous cell carcinoma of the head and neck (SCCHN), the 6th most common malignancy in the world, is associated with smoking and has a low 5-year survival rate. Various changes have been described at different stages of SCCHN tumour development, including overexpression of p63, a protein important for development of normal epidermal structures. p63 has been suggested to activate beta-catenin, and nuclear accumulation of beta-catenin is an important event in many cancers. Elevated COX-2 activity and overexpression of EGFR protein has been shown in a variety of human cancers, including SCCHN. An important question for the pathogenesis of SCCHN is when the genetic changes take place during the natural course of the disease, and whether they appear in clinically normal oral mucosa to predispose tumour development. We mapped the expression of p63, COX-2, EGFR, beta-catenin, and PP2A in oral mucosa from smokers/non-smokers and from patients with SCCHN. We also considered if changes occurring in tumours are present in the clinically normal tissue adjacent to the tumour. No direct influence of heavy smoking on the levels of the proteins studied could be seen. Tumours and clinically normal non-neoplastic tissue from SCCHN patients showed increased expression of COX-2 and PP2A. Interestingly, non-neoplastic tissue adjacent to SCCHN also showed increased beta-catenin, although this was not seen in tumours. The data support the notion that pre-existing alterations in clinically normal epithelium exist in patients with SCCHN and could be important for the pathogenesis of the disease and for local recurrences. PMID- 16273223 TI - Cytotoxicity of the polyamine oxidase inactivator MDL 72527 to cancer cells: comparison with a saturated structural analogue. AB - MDL 72527 (N1,N4-di-2,3-butadienyl-1,4-butanediamine) is a selective inactivator of polyamine oxidase with therapeutic potential. However, the development of lethal toxic effects due to prevention of spermine degradation is a considerable disadvantage of the compound. Since the cytotoxicity of MDL 72527 was postulated to be independent of its anti-polyamine oxidase activity, its cytotoxicity to cancer cells was compared with that of a close analogue that is devoid of structural features enabling mechanism-based inactivation of polyamine oxidase. N1,N4-di-n-butyl-1,4-butanediamine proved to be a cytotoxic agent of considerable potency, which induces mainly non-apoptotic cell death, whereas MDL 72527 causes under identical conditions both, apoptotic and non-apoptotic cell death. The sensitivity of cells to both compounds is presumably dependent of their glutathione content. PMID- 16273224 TI - Epithelial-mesenchymal transition in gastric cancer (Review). AB - Endoscopic mucosal resection (EMR), endoscopic submucosal dissection (ESD), surgical gastrectomy, and chemotherapy are therapeutic options of gastric cancer; how-ever, prognosis of advanced gastric cancer patients is still poor. Gastric cancer cells with fibroblastoid morphological changes show increased motility and invasiveness due to decreased cell-cell adhesion, which are reminiscent of epithelial-mesenchymal transition (EMT) during embryonic development. Here, EMT signaling networks in gastric cancer were reviewed. E-cadherin at adherens junction is a key molecular target of EMT. CDH1 gene at human chromosome 16q22.1 encodes E-cadherin. Familial diffuse type gastric cancer occurs due to germ-line mutations of the CDH1 gene. Down-regulation of E-cadherin function due to mutation, deletion, CpG hyper-methylation, and SNAIL (SNAI1)- or SIP1-mediated transcriptional repression of the CDH1 gene leads to EMT in gastric cancer. Amplification of ERBB2, MET, FGFR2, PIK3CA, AKT1 genes, up-regulation of WNT2, WNT2B, WNT8B, and down-regulation of SFRP1 lead to EMT in gastric cancer through GSK3beta inhibition and following SNAIL-mediated CDH1 repression. Claudin (CLDN) and PAR3/PAR6/aPKC complex at tight junction are other key molecular targets of EMT. CLDN23 gene is down-regulated in intestinal type gastric cancer. Down regulation of PAR3/PAR6/aPKC complex also leads to EMT. Single nucleotide polymorphisms (SNPs) and copy number polymorphisms (CNPs) of genes encoding EMT signaling molecules will be identified as novel risk factors of gastric cancer. In addition, antibodies, RNAi compounds, and small molecular inhibitors for EMT signaling molecules will be developed as novel therapeutic agents for gastric cancer. Personalized medicine based on the combination of genetic screening and novel therapeutic agents could dramatically improve the prognosis of gastric cancer patients in the future. PMID- 16273225 TI - Enhanced induction of prostatic dysplasia and carcinoma in Noble rat model by combination of neonatal estrogen exposure and hormonal treatments at adulthood. AB - Estrogens have been implicated to play certain but yet undefined roles in the normal and neoplastic growth of prostate gland. Studies of perinatal exposure in rodents demonstrate that effects of perinatal estrogenization are permanent and carcinogenic in prostate gland. In the Noble (Nb) rat model, prostatic dysplasia and neoplastic lesions can be induced by a chronic treatment with both testosterone and estrogen at adulthood. However, by this conventional protocol, neoplastic lesions are mostly confined to the lateral (LP) and ventral (VP) prostates, while gross prostatic tumors are rarely induced. Based on these two experimental models, we developed a modified treatment protocol for the enhancement of prostate cancer induction in Nb rat model by combining neonatal estrogen exposure of male offspring followed by the hormonal treatment at adulthood (NeoE + T-E2). Using this modified protocol, we were able to induce more extensive development of neoplastic lesions in all three prostatic lobes and also gross tumors at relatively high incidence within 6-9 months. Western blottings and immunohistochemistry showed that ERalpha expression was increased in the hypertrophic peri-acinar and -ductal smooth muscle cells while ERbeta and AR expressions are markedly decreased in dysplastic and neoplastic lesions in NeoE + T-E2-treated prostates. Immunohistochemistry showed that expression of three tumor suppressors (BRCA2, PTEN, and Rap1) and tubulin-alpha are markedly decreased in dysplastic and neoplastic lesions. In addition, loss of expression of smooth muscle differentiation markers (desmin, alpha-actin, and vinculin) and defects of basement membranes were also seen in the reactive stroma. These results suggest that exposure to high levels of estrogens, either endogenous or exogenous, in early life could play a role in the development of prostate cancer in later life. PMID- 16273226 TI - Presence of MLH1 protein aggravates the potential of the HSP90 inhibitor radicicol to sensitize tumor cells to cisplatin. AB - MLH1 is one of five proteins crucial to DNA mismatch repair (MMR) function the loss of which is associated with a cisplatin resistance phenotype in tumor cells. An experimental approach was designed to determine whether the presence or absence of MLH1 affects the potential of radicicol to increase the sensitivity of tumor cells to cisplatin and oxaliplatin, and whether perhaps radicicol increases sensitivity to cisplatin specifically in cisplatin-resistant, MLH1-deficient cells. Radicicol is a novel specific inhibitor for heat shock protein 90 (HSP90) and structurally unrelated to geldanamycin. Clonogenic data demonstrated that sublethal concentrations of radicicol increased the sensitivity to cisplatin and to oxaliplatin in both MLH1-proficient cells and MLH1-deficient cells. Notably, the radicicol-imposed increase in sensitivity to cisplatin was up to 1.6-fold higher in MLH1-proficient cells than in MLH1-deficient cells, whereas no difference in the extent of the increase in sensitivity between the two sublines was observed for oxaliplatin. This indicates that the presence of MLH1 protein aggravates the radicicol-imposed increase in sensitivity of cells to cisplatin but not to oxaliplatin. However, the increases in platinum drug sensitivity imposed by radicicol observed in the clonogenic assay were not accompanied by reproducible alterations in the susceptibility to apoptosis and to changes in cell cycling. Although not conclusive at this point, the results seem to argue against radicicol as a means to selectively re-sensitize cisplatin-resistant, MLH1-deficient tumor cells to this drug. But they may point to a possible functional relationship between HSP90 and MLH1, where HSP90 might affect the function of MLH1 in a way that this leads to the counter-regulation of cytotoxic pathways initiated by MMR as a consequence of the presence of DNA damage introduced by cisplatin. PMID- 16273227 TI - The role of combined allelic imbalance and mutations of p53 in tumor progression and survival following surgery for colorectal carcinoma. AB - The role of p53 mutations in disease progression and survival of colorectal cancer is unclear, since numerous studies have reported different conclusions. However, few reports, if any, have evaluated disease progression and survival in relationship to 'functional' and 'non-functional' p53 status defined by genetic and molecular indications. Malignant colorectal tumors, from 72 unselected patients who underwent primary and potentially curative elective tumor resections, were either classified as p53 functional (p53+/+, p53+/-) or non functional (p53-/-) based on DNA sequence analysis of all p53 exons, including determination of allelic imbalance of p53 (LOH), according to four DNA markers; 2 within the coding gene and two markers in the immediate flanking regions of p53. Tumor frequency of microsatellite instability was also analyzed according to Dukes' A-D stages. Dukes' staging predicted survival as expected, while the conceptual p53 status, functional p53 vs non-functional p53, did not clear-cut predict disease specific survival. p53 mutations alone or allelic imbalance inside the reading frame of the gene were unpredictive of survival, while allelic imbalance downstream of p53 predicted reduced survival (p < 0.05). The present study demonstrates that base mutations in combination with allelic imbalance within the reading frame of p53 do not predict survival or progression of colorectal cancer, while allelic imbalance upstream coding parts of the gene predicted disease-specific survival in univariate analysis. Thus, structural alterations within the gene seem less important than alterations in regions with potential control elements. PMID- 16273228 TI - Contribution of reactive oxygen species and caspase-3 to apoptosis and attenuated ICAM-1 expression by paclitaxel-treated MDA-MB-435 breast carcinoma cells. AB - Paclitaxel is a microtubule-stabilizing and apoptosis-inducing drug that is commonly used to treat metastatic breast cancer, although the mechanism of paclitaxel-induced apoptosis remains incompletely understood. Furthermore, adhesion molecule expression is attenuated on mouse mastocytoma and human leukemia cells that survive short-term culture in the presence of paclitaxel. In the present study we show that MDA-MB-435 human breast carcinoma cells that survived culture for 72 h in the presence of submaximal cytotoxic concentrations of paclitaxel (0.02 and 0.01 microg/ml) showed decreased expression of the adhesion molecule ICAM-1. Paclitaxel treatment of MDA-MB-435 cells was associated with the generation of reactive oxygen species (ROS), dissipation of mitochondrial transmembrane potential, and the activation of caspase-3. The antioxidant glutathione protected MDA-MB-435 cells from paclitaxel-induced cytotoxicity and reduced ICAM-1 expression. In addition, a selective inhibitor of caspase-3 (Z-DEVD-FMK), as well as a pan-caspase inhibitor (Z-VAD-FMK), partially prevented the decrease in ICAM-1 expression observed following paclitaxel treatment, but did not protect against paclitaxel-induced cytotoxicity. We conclude that the paclitaxel-induced reduction in ICAM-1 expression by MDA-MB-435 breast carcinoma cells is both ROS- and caspase-dependent, whereas paclitaxel induced cytotoxicity is ROS-dependent and does not involve caspases. Decreased ICAM-1 expression by breast carcinoma cells that survive paclitaxel treatment may negatively impact on cytotoxic lymphocyte-mediated destruction of paclitaxel resistant breast cancer cells in the context of chemo-immunotherapy or chemo adoptive immunotherapy. PMID- 16273229 TI - Differential gene expression of sulindac-treated human breast epithelial cells. AB - Breast cancer is the most common malignancy and the second major cause of cancer related deaths among women in the United States. Recent advances in the molecular genetics of breast cancer have identified various genes associated with tumorigenesis. There is evidence that non-steroidal anti-inflammatory drugs, e.g. sulindac, have some anti-proliferative effects on various tumors involving altered p53 function. Most of these studies have been performed with various human colon carcinoma cell lines and few of them focus on non-malignant proliferative human mammary epithelial cell lines. Therefore, the present study was undertaken to analyze the differentially expressed genes of the p53 signaling pathway by means of a gene array for the immortalized human breast epithelial cell line, MCF-10F, treated with sulindac. Out of the total 96 genes, only 17 were altered by the drug treatment. Among these 17 genes, 6 showed significant alteration (Q > 2.0), whereas 11 genes showed moderate alterations. Altered genes included BRCA1 associated protein-1 [ubiquitin carboxy-terminal hydrolase (bap1)]; cell division cycle 2, G1 to S and G2 to M [cdk1(cdc2)]; and DNA-damage inducible transcript 1 (gadd45), which were down-regulated. However, N-myc gene 1 (rtp), promyelocytic leukemia (pml), and nuclear factor of kappa-light polypeptide gene enhancer in B-cell 3 and p65 [avian (rel A)] were up-regulated. Northern blot analysis confirmed some of these alterations. The alteration of p53 signaling pathway gene markers by sulindac treatment can give us valuable information about the response to drug treatments in a proliferative cell population. PMID- 16273230 TI - Conjugated linoleic acid induces monocytic differentiation of murine myeloid leukemia cells. AB - Conjugated linoleic acid (CLA) refers to a group of naturally occurring positional and geometrical conjugated dienoic isomers of linoleic acid (C18:2), of which the cis-9,trans-11 (c9,t11) and trans-10,cis-12 (t10,c12) isomers predominate. Accumulating evidence has demonstrated that CLA isomers are capable of inhibiting the growth of a variety of cancer cell lines in vitro; however, their modulatory effects on the proliferation and differentiation of myeloid leukemia cells remain poorly understood. In the present study, CLA was shown to inhibit the proliferation of murine myeloid leukemia WEHI-3B JCS cells in a dose- and time-dependent manner. Morphological, flow cytometric and functional analyses revealed that CLA induced the differentiation of WEHI-3B JCS cells into matured macrophage-like cells, as indicated by increases in the cytoplasm:nucleus ratio and vacuolation, the expression of macrophage differentiation antigens (Mac-1 and F4/80) and the enhanced monocytic serine esterase activity of CLA-treated WEHI-3B JCS cells. RT-PCR analysis showed that CLA up-regulated the expression of TNF alpha, IL-1beta and IFN-gamma genes in WEHI-3B JCS cells, which had previously been shown to play an important role in triggering the differentiation of myeloid leukemia cells. Moreover, CLA-treated WEHI-3B JCS cells had also shown reduced tumorigenicity in vivo. Collectively, our results indicate that CLA might exert its growth-inhibitory effects on myeloid leukemia cells by triggering their terminal differentiation, which is mediated, at least in part, by modulation of the cytokine gene expression in the leukemia cells. PMID- 16273231 TI - Anti-angiogenic effects of a nutrient mixture on human umbilical vein endothelial cells. AB - Matrix metalloproteinases (MMPs) have been recognized as key players in the degradation of the extracellular matrix (ECM) by migration and proliferation of endothelial cells and their subsequent invasion of the underlying stroma. The prevention of ECM degradation through the inhibition of MMP activity has been shown to be a promising therapeutic approach to block the invasion that occurs during angiogenesis. In previous studies, we demonstrated the anti-tumor effect of a nutrient mixture (NM) containing ascorbic acid, lysine, proline, green tea extract, arginine, N-acetyl cysteine, selenium, copper and manganese on various tumor cell lines in vivo and in vitro. The aim of the present study was to determine whether this mixture has anti-angiogenic effects on human umbilical vein endothelial cells (HUVECs). At near confluence, the HUVEC cell cultures were tested with NM at 0, 10, 50, 100, 500, and 1000 microg/ml in triplicate at each dose for proliferation, migration, MMP expression, and invasion. Cell proliferation was evaluated by MTT assay, invasion potential by Matrigel invasion, MMP expression by gelatinase zymography, and cell migration by a 2 mm wide scratch in plates. For tube formation, HUVECs were cultured in previously polymerized Matrigel. NM inhibited HUVEC migration, MMP expression and invasion through Matrigel in a dose-dependent manner. Zymography showed a dose-dependent inhibition of MMP-2 expression with virtual total inhibition at a 500 microg/ml concentration. Invasion through Matrigel was totally inhibited at 500 microg/ml NM. NM reduced cell migration by scratch test in a dose-dependent fashion with total inhibition at a 500 microg/ml concentration. NM also inhibited the tube formation of HUVECs, but did not significantly inhibit cell proliferation. These results together with our earlier findings suggest that NM is a relatively non toxic formulation with anti-angiogenic effects, such as inhibiting vascular tube formation and endothelial cell invasion and migration. PMID- 16273232 TI - Use of granulocyte colony-stimulating factor: a survey among Italian medical oncologists. AB - In October 2003, the Italian Association of Medical Oncology (AIOM) published its own guidelines on the use of granulocyte colony-stimulating factor (G-CSF). The present survey was conducted during the same period with the aim of collecting data on the current use of G-CSF to provide a starting point for future evaluations of the implementation of AIOM guidelines. From October 2003 to January 2004, 1591 AIOM members were asked to complete a questionnaire based on specific clinical scenarios, regarding the use of G-CSF for primary and secondary prophylaxis and treatment of neutropenia. The rate of response was 22%. For primary prophylaxis, the majority of physicians avoid using G-CSF, with no difference in cases of adjuvant, curative or palliative chemotherapy (CT). In fact, 67.2% to 74.9% would 'rarely or never' use G-CSF in the proposed clinical scenarios. In chemosensitive tumors, rather than reducing CT doses, 55.7% would use G-CSF as a secondary prophylaxis after afebrile neutropenia (AN), and 68.8% after febrile neutropenia (FN). In elderly patients experiencing FN, 35.7% would reduce the adjuvant CT doses and 23.1% would change the regimen. Most oncologists would use G-CSF to treat neutropenia, and the median duration of G-CSF treatment is less than 1 week and would depend on neutrophil count. Our survey shows that Italian oncologists are particularly oriented towards the use of G-CSF in clinical practice to maintain the CT dose intensity, and are sensitive to the prevention and treatment of not only FN, but also AN. Finally, Italian medical oncologists appear to be very cautious in introducing G-CSF when treating elderly patients. PMID- 16273234 TI - Radiosensitivity of hypoxic and proliferating clonogen in a human lung cancer grown in nude mice. AB - Using cultured and nude mouse tumor cells (IA) derived from a human lung cancer, we studied their radiosensitivity by focusing attention on the dynamics of tumor clonogens. The movement of clonogens in the regrowing IA tumor after irradiation can be divided into three phases: first, the early and rapid survival recovery (PLD repair) phase; second, the delay phase involving a certain lag in survival change; and third, the repopulation phase consisting of two stages: the anoxic repopulation before angiogenesis and the hypoxic repopulation in the presence of a poorly developed vascular network. Clonogens in a regrowing tumor after irradiation were found to actively proliferate even in an anoxic environment before angiogenesis and under the hypoxic conditions prevailing after the formation of a tumor with a poorly developed vascular system. This re-grown tumor was found to be more radioresistant than a sham-treated control tumor. It is believed that these clonogens are genetically selected under hypoxic conditions throughout the process of tumor growth and regrowth, and may be primarily involved in tumor recurrence or accelerated repopulation in fractionated irradiation. PMID- 16273233 TI - The expression of CD97EGF and its ligand CD55 on marginal epithelium is related to higher stage and depth of tumor invasion of gastric carcinomas. AB - CD97EGF is a member of the EGF-TM7 family of class II seven-transmembrane (7TM), and its cellular ligand CD55 (also known as decay accelerating factor; DAF) protects host cells from complement attack. To determine whether the expression levels of these two molecules are correlated with the clinicopathological features of gastric carcinomas, a total of 35 gastric carcinomas and their corresponding margins and normal specimens were investigated by RT-PCR, Western blot analysis and immunohistochemistry. Transcript levels of CD97EGFand CD55 were higher in tumors than those in the margin and normal epithelial mucous tissues (P<0.05). However, the expression levels of CD97EGF and CD55 mRNA had no correlation with the clinicopathological features of gastric carcinoma patients. All three groups of specimens were immunoreactive for CD97EGF and the CD55 protein. Strong and specific immunoreactivities of CD97EGF were located in the mucosal epithelia of the marginal basal membrane. Expression of CD97(EGF) in the margins showed a marked difference between the depth of tumor invasion T1 and T2, 3 and 4, and stages I and II/III/IV of gastric carcinomas (P<0.05). The expression of CD55 protein was highly correlated with CD97EGF (R=0.6483, P<0.001). Western blot analysis confirmed the expression and distribution patterns of CD97EGF and CD55. Our findings suggest that CD97EGF may play a role in the development and invasion of gastric carcinomas by binding its cellular ligand CD55. Detection of the CD97EGF expression in the tumor margin could be referred to as the molecular edge of gastric carcinomas. PMID- 16273235 TI - Genomic alterations detected by comparative genomic hybridization in primary lung adenocarcinomas with special reference to the relationship with DNA ploidy. AB - To clarify the clinicopathological and biological significance of genomic alterations in pulmonary adenocarcinomas, we examined chromosomal DNA sequence copy number aberrations (DSCNAs) and DNA ploidy in 42 surgically resected specimens by comparative genomic hybridization (CGH) and laser scanning cytometry (LSC), respectively. The number of DSCNA did not affect the size of carcinoma or number of nodal metastasis. More than 60% of carcinomas showed a 1q21-23 gain or 13q21 loss. Gains of 9q22-33 and 10q26-qter and a loss of 15q14-qter were significantly associated with nodal metastasis (p<0.05). Of 42 pulmonary adenocarcinomas, 32 (76%) showed DNA aneuploidy. The number of DSCNAs in aneuploid carcinomas was larger than that in diploid carcinomas (p<0.01). Our results suggest that diploid and aneuploid types are cytogenetically different in pulmonary adenocarcinomas and aneuploid carcinomas are genetically more unstable and aggressive than diploid carcinomas. PMID- 16273236 TI - Alterations and correlations of the components in the Wnt signaling pathway and its target genes in breast cancer. AB - Both cyclin D1 and c-myc are key molecules in breast cancer carcinogenesis, and their transcriptional level and stability are regulated through several signaling pathways, including the Wnt signaling pathway. We performed immunohistochemical and mutational analyses of Wnt signaling components to investigate the association of Wnt signaling alterations with breast cancer carcinogenesis using 49 surgically resected primary breast cancer samples. Positive staining of cyclin D1 and c-myc was observed in 55.1% and 30.6% of the 49 breast cancer samples, respectively. Aberrant cytoplasmic expression of beta-catenin, which indicates the existence of alterations in the Wnt signaling pathway, was observed in 38.8% of breast cancer samples, though no mutation was found in the beta-catenin and Axin 1 genes. Reduced expression of APC was observed in 34.7% of samples. Statistical analysis revealed strong correlations between overexpression of beta catenin and that of cyclin D1 and c-myc (p=0.0001 and 0.0117, respectively). Furthermore, overexpression of beta-catenin was significantly correlated with reduced expression of APC (p=0.0127). Wnt signaling alterations were frequently observed in breast cancer from the results of beta-catenin immunohistochemistry, although no mutation in the components of the Wnt signaling pathway was found in the present study. Based on the statistical analyses, we speculated that reduced expression of APC leads to overexpression of beta-catenin, and aberrant expression of cyclin D1 and c-myc mainly depends on alterations in the Wnt signaling pathway in breast cancer. PMID- 16273237 TI - Plasminogen activator inhibitor-1 (PAI-1) gene transfection inhibits the liver metastasis of pancreatic cancer by preventing angiogenesis. AB - Plasminogen activator inhibitor-1 (PAI-1) is a unique type of serine protease inhibitor and one of the key regulators of tumor invasion and metastasis. The purpose of this study was to elucidate the effect of PAI-1 gene transfection on liver metastasis and its mechanism by using the human high liver metastasis pancreatic cancer cell line, SW1990. PAI-1-transfected SW1990 (SW/PAI-1) produced a significantly higher level of PAI-1 in supernatant than parental cells. While no difference was observed for the production of u-PA and u-PA activity in the supernatant, cell proliferation of SW/PAI-1 was slightly suppressed on the 7th day of incubation compared to parental cells. Cellular invasion, in vivo tumorigenesis in xenograft and liver metastasis were significantly suppressed in SW/PAI-1 cells compared to parental cells. The angiogenesis of xenograft by detecting microvascular density and the production of metastasis-related factors, such as VEGF and TGF-beta1, were also decreased in SW/PAI-1 cells. These findings suggested that PAI-1 gene transfection might have the ability to prevent the liver metastasis of pancreatic cancer by modulating angiogenesis. PMID- 16273238 TI - Alterations of p53, cyclin D1 and pRB expression in the carcinogenesis of esophageal squamous cell carcinoma. AB - Many molecular alterations occur in esophageal carcinogenesis; however, little is known about the molecular genetic events responsible for the development of carcinoma. We investigated the expression of ki67, p53, cyclin D1 and pRB in 105 biopsy specimens using immunohistochemistry from iodine unstained lesions as indicators of carcinogenesis of the esophagus. Also, the genetic alternation of esophageal dysplasia from patients with accompanying esophageal squamous cell carcinoma (ESCC) was examined to study the evidence for field carcinogenesis in the esophagus. The expression of p53, cyclin D1 and pRB was detected in 31, 0 and 51.7% respectively of mild dysplasia; 40, 0 and 70% of moderate dysplasia; 40, 20 and 70% of severe dysplasia; and 48, 32 and 80% of carcinoma specimens. p53 expression was significantly increased in mild dysplasia, whereas cyclin D1 and pRB expression were significantly increased in carcinoma as compared to both normal epithelium and esophagitis. The ki67 LI and the rate of p53 expression were significantly higher in dysplasia with ESCC than in dysplasia without ESCC. Ki67, p53, cyclin D1 and pRB expression may be useful biomarkers for assessing the risk of developing esophageal cancer. Dysplasia observed at screening for secondary lesions has a highly malignant potential and careful follow-up studies are required. PMID- 16273240 TI - Pathomorphological study on small nodular lesions in hepatitis C virus-related cirrhosis. AB - We examined pathomorphologically small nodular lesions in hepatitis C virus related cirrhosis (type C cirrhosis). Small nodular lesions seen in the non cancerous areas of 128 consecutively resected hepatocellular carcinomas (HCCs) associated with type C cirrhosis were selected for the study. From these, 18 large regenerative nodules (LRNs), 14 low-grade dysplastic nodules (LDNs), 10 high-grade DNs (HDNs), and 12 well-differentiated HCCs were detected. The diameters of HDN (11.7+/-4.3 mm) and well-differentiated HCC (12.3+/-3.5) were significantly larger than those of LRN (8.1+/-1.0) and LDN (8.6+/-2.7). Fatty change was not seen in LRNs and LDNs, while it was found in 4 HDNs (40%) and 8 well-differentiated HCCs (75%). Iron deposits were found in 2 LRNs (11%), 5 LDNs (36%), and 3 HDNs (30%), but not in the well-differentiated HCCs. The number of Kupffer cells in LDN and HDN was increased compared with that in the surrounding regenerative nodules. Cell density and the degree of neovascularization were well correlated with the progression from DN to HCC. The lack of neovascularization supports the concept that LRN has no neoplastic potency and can be regarded as the morphologic marker in the differentiation of LRN from DN. PMID- 16273239 TI - Cyclin B1 expression is an independent prognostic marker for poor outcome in diffuse large B-cell lymphoma. AB - Diffuse large B-cell lymphoma (DLBCL) is the most frequent non-Hodgkin's lymphoma in the Western world; it is characterized by marked genetic, morphological and clinical heterogeneity. The identification of prognostic markers could help to develop risk-adapted treatment strategies. As proliferation of cells is essential for tumour growth, analysis of the cell cycle and its individual phases might give additional information on tumour progression and clinical behaviour. To investigate the prognostic value of proteins specifically related to the cell cycle phases S, G2 and M in DLBCL, the expression of cyclin A as a marker of S phase and cyclin B1 as a G2/M phase marker were analysed in combination with other clinicopathological parameters in a large cohort of patients. Expression of cyclin B1 and cyclin A were determined by immunohistochemistry using tissue microarray methodology. Immunoreactivity for cyclin B1 and cyclin A was correlated with clinical data using a two-sided Fisher's exact test. Impact on overall survival was analysed by the Kaplan-Meier method. A negative prognostic impact was found for expression of cyclin B1. Nuclear and/or cytoplasmic staining in > or = 1% of tumour cells was significantly associated with shorter overall survival in the multivariate analysis (p=0.008). Furthermore, the prognostic impact of cyclin B1 expression was independent of the tumour stage. No prognostic significance was found for expression of cyclin A. In conclusion, this study demonstrates the independent prognostic value of an expression of cyclin B1 in DLBCL and proposes its evaluation as a prognostic marker in the assessment of this entity which is easily applicable in daily routine practice. PMID- 16273241 TI - Treatment with hydroxyurea and tyrphostin-1 significantly improves the transduction efficiency of recombinant adeno-associated viruses in human cancer cells. AB - To enhance the transduction efficiency (TE) of a recombinant adeno-associated virus 2 (rAAV2) in human cancer cells, we examined the combined effects of various chemicals known to influence the rAAV2 transduction process at distinct steps. Among the agents tested were trichostatin A, a histone deacetylase inhibitor, MG-132, a proteosome inhibitor, the genotoxic agents hydroxyurea, aphidicolin, etoposide and camptothecin, and tyrphostin-1, an epidermal growth factor receptor inhibitor. During or after chemical treatment, various human cancer cells were infected with rAAV2 expressing beta-galactosidase. Treatment with hydroxy-urea or etoposide plus tyrphostin-1 dramatically increased the TE in most cell lines. The combination of hydroxyurea plus tyrphostin-1 increased TE to 37.7+/-7.9%, 32.8+/-2.0% and 31.8+/-2.1% in SK-Hep1, HeLa, and HCT116 cells, respectively. In addition, following rAAV2 infection and treatment with hydroxyurea plus tyrphostin-1, long-term transgene expression was observed for up to 6 months, with no damage to the transduced cells. These results indicate that rAAV2 transgene expression can be significantly enhanced by a combination of chemical agents with distinct activity and prolonged gene expression can occur following rAAV2 gene transfer into human cancer cells. PMID- 16273242 TI - Exploration of genetic alterations in human endometrial cancer and melanoma: distinct tumorigenic pathways that share a frequent abnormal PI3K/AKT cascade. AB - Mutations of RAS, RAF, and PTEN, all important members of the RAS/MAPK and PI3K/AKT cascades, are reported in a variety of human tumors, including melanomas and endometrial cancer. In endometrial cancer, mutually exclusive mutations of PTEN and KRAS have been reported. On the other hand, mutation of BRAF is highly frequent, and mutually exclusive mutations of BRAF and NRAS have also been reported in melanomas. In this study, we elucidated the involvement of the up regulation of RAS/MAPK and PI3K/AKT cascades in the pathogenesis of endometrial cancer and melanoma by analyzing the genes and molecules in these cascades. Twelve cell lines, six melanoma and six endometrial cancer, were analyzed; 4 (67%) of the 6 melanomas had gene mutations in the RAS/MAPK cascade, and a decrease or loss of PTEN expression was also observed. These results suggested that simultaneous up-regulations in these two cascades play important roles in carcinogenesis of melanocytes. However, no activation of AKT by phosphorylation was observed. On the other hand, 4 (67%) of the 6 endometrial cancer cell lines had mutually exclusive up-regulations in these cascades. However, two cell lines with up-regulation of the PI3K/AKT cascade also had up-regulation in the RAS/MAPK cascade induced by inactivation of DUSP6. These results suggest that simultaneous up-regulation of RAS/MAPK and PI3K/AKT cascades are crucial events in the pathogenesis of melanocytes, whereas up-regulation of either the RAS/MAPK or PI3K/AKT cascade is crucial for the majority of endometrial cancers. PMID- 16273243 TI - The influence of oral tumor cell proliferation activity and membrane potential on the transfection efficiency of a cationic liposome. AB - The transfection efficiency of cationic liposomes varies according to cell type, but the specific cellular characteristics that affect transfection efficiency have not yet been defined. We investigated whether the transfection efficiency of cationic liposomes correlates with cell proliferation activity or cell membrane potential in oral malignant melanoma (HMG) and oral osteosarcoma cell lines (HOSM 1 and HOSM-2). The cell membrane potential was assessed by uptake of a cationic probe. Three oral tumor cell lines were exposed to a cationic liposome complexed with a beta-galactosidase expression plasmid, and beta-galactosidase expression was compared. Cell proliferation was about 2-fold higher in HOSM-1 cells than in HMG cells. The cell membrane potential in HMG and HOSM-1 cells was comparable, while the membrane potential in HOSM-2 cells was 1.6-fold higher. beta galactosidase expression was measured by X-Gal staining in 7.0% of HMG, 17.0% of HOSM-1 and 11.5% of HOSM-2 cells. The present study demonstrates that gene therapy with cationic liposomes may be a promising new strategy for treatment of oral malignant melanoma and osteosarcoma. In addition, the transfection efficiency of cationic liposomes appears to be influenced by cell proliferation activity, but not cell membrane potential. PMID- 16273244 TI - S100A2 expression as a predictive marker for late cervical metastasis in stage I and II invasive squamous cell carcinoma of the oral cavity. AB - The purpose of this study was to discover whether S100A2 expression is associated with late cervical metastasis in patients with stage I and II invasive squamous cell carcinoma of the oral cavity. We retrospectively investigated the clinicopathological parameters and S100A2 expression in surgical specimens taken from 52 patients with T1-2N0M0 invasive squamous cell carcinoma of the oral cavity (OSCC) who had not undergone elective neck dissection or irradiation to the neck. All of the clinicopathological factors and S100A2 expression were compared in terms of late cervical metastasis. In univariate analysis, late cervical metastasis correlated with poor overall survival. A higher rate of late cervical metastasis was observed in patients with S100A2-negative tumors than those with S100A2-positive tumors. Multivariate analysis on late cervical metastasis revealed that S100A2 expression was demonstrated to be the only independent factor for late cervical metastasis. Our results indicate that patients with stage I or II invasive OSCC without S100A2 expression should be considered a high-risk group for late cervical metastasis when a wait-and-see policy for the neck is being considered. PMID- 16273245 TI - Glucose transporter-1 expression in the thyroid gland: clinicopathological significance for papillary carcinoma. AB - Glucose transporter-1 (GLUT-1) expression was immunohistochemically analysed in a total of 268 cases of thyroid gland disease, including 129 cases of papillary carcinoma (PC), 60 cases of follicular carcinoma (FC), 57 cases of follicular adenoma, and 22 cases of adenomatous goitre. Seventy-one percent (91/129) of PC cases showed GLUT-1 expression, semi-quantitatively evaluated as: +, 21 cases (16%); 2+, 37 cases (29%); 3+, 33 cases (26%); and negative, 38 cases (29%). These positive cases were divided into two groups: 'membrane-like' pattern in 24 cases (19%), and 'cytoplasm-predominance' pattern in 67 cases (52%). GLUT-1 expression was observed in 5% (3/60) of FC cases, but all follicular adenomas and adenomatous goitres were negative for GLUT-1 (PC vs. FC, p<0.0001). Membrane-like expression was observed more frequently in non-organ-confined PCs (pT4) than in organ-confined PCs (pT1, 2, and 3) (p=0.0056). Seventy-five percent (18/24) of PC cases showing membrane-like expression were non-organ-confined. The membrane-like pattern was observed more frequently in PCs with lymph node (LN) metastasis compared to those without (p=0.0036). Ninety-two percent (22/24) of PC cases showing the membrane-like pattern were not organ-confined. Semi-quantitative analysis of glut-1 mRNA by RT-PCR showed a tendency toward higher expression in PCs compared to FCs, follicular adenomas and adenomatous goitres, and the mRNA expression in PCs with a membrane-like pattern was higher than those showing cytoplasm-predominance. We concluded that: 1) GLUT-1 is immunohistochemically useful in distinguishing PC from FC and benign diseases; 2) GLUT-1 may play an important role in the advancement of PC and LN metastasis, and its membrane-like expression is of more clinical significance than the cytoplasm-predominance pattern; and 3) glut-1 mRNA expression corresponds with the immunohistochemical expression profile. PMID- 16273246 TI - Recurrence and 5-FU sensitivity of stage II/III node-positive gastric cancer with occult neoplastic cells in lymph node sinuses. AB - The 5-year overall survival (OS) rates for patients without occult neoplastic cells (ONCs) were 43.0% in stage II (n=15), 52.2% in stage III (n=23), and 48.5% for stages II and III combined (n=38). For ONC-positive patients, the 5-year OS rates were 44.4% in stage II (n=7; p=0.88322), 11.3% in stage III (n=30; p=0.0006), and 17.5% for stages II and III combined (n=37; p=0.0019). Among the ONC(+) recurrence group (75.7%, 28/37), 42.9% (12/28) showed high TS expression in metastatic lymph nodes and 57.1% (16/28) showed low TS expression. In the case of DPD expression, 32.1% (9/28) showed high expression and 67.9% (19/28) showed low expression. Among the ONC(+) non-recurrence group (24.3%, 9/37), 66.7% (6/9) showed high TS expression and 33.3% (3/9) showed low TS expression, while high and low DPD expression was seen in 22.2% (2/9) and 77.8% (7/9), respectively. A combination of high TS and low DPD expression was found in 32.1% (9/28) of the recurrence group vs. 66.7% (6/9) of the non-recurrence group (p=0.070). These results suggest that ONCs are associated with OS. Unlike the non-recurrence group, the ONC(+) patients with recurrence of stage II/III node-positive gastric cancer are unlikely to respond to treatment with 5-FU + LV and may need combination chemotherapy based on L-OHP and/or CPT-11. PMID- 16273247 TI - Two different global gene expression profiles in cancer cell lines established from etiologically different oral carcinomas. AB - cDNA arrays were used to characterize the gene expression profiles in 6 oral carcinoma cell lines (UT-SCC-10, UT-SCC-14, UT-SCC-37, UT-SCC-54A and UT-SCC-54B, UT-SCC-74) established from 5 patients with different etiological backgrounds, including young patients, classical risk factors and lichen-derived lesions. In addition, 2 human papillomavirus (HPV)-positive cell lines (hypophraryngeal cancer and HPV16 E6/E7-transformed oral keratinocytes) were similarly tested. Two distinct global gene expression profiles with down-regulated and up-regulated patterns were identified, which closely related to the etiologic backgrounds of the primary tumors. Typically in cluster analysis, interferon or interferon related genes and T- and B-lymphocyte-related genes were up-regulated in lichen derived carcinoma cell lines. Common to all carcinoma cell lines were 6 genes, which were up- or down-regulated (IgC mu heavy chain constant region, semaphorin, T-cell growth factor, cAMP-dependent protein kinase beta-catalytic subunit, desmocollin 1A/1B precursor and recA-like protein HsRad51). In HPV-positive cell lines, 13 genes were identified with similar down-regulation as shown in our previous studies on HPV-positive genital cell lines. Importantly, all of these genes were also down-regulated in 3 of the 6 oral cancer cell lines. These data suggest that oral carcinomas with different etiological backgrounds can be distinguished by their different global gene expression patterns. PMID- 16273248 TI - Up-regulation of PINCH in the stroma of oral squamous cell carcinoma predicts nodal metastasis. AB - Particularly interesting new cysteine-histidine rich protein (PINCH), an adapter protein involved in integrin and growth factor signalling, is up-regulated in the stroma of colorectal, breast, prostate, lung and skin cancer. Strong stromal immunostaining for PINCH is an independent prognostic indicator for reduced survival in colorectal cancer, suggesting that PINCH is involved in the signalling that promotes tumour progression. Since no study on PINCH has been carried out in oral squamous cell carcinoma (OSCC), this study aimed to determine PINCH expression in OSCC and its clinicopathological significance. PINCH protein expression was examined by immunohistochemistry in 20 normal oral mucosa and in 57 OSCC specimens. The frequency of strong PINCH immunostaining was higher in tumour-associated stroma of OSCC (37%) as compared to normal oral mucosa (10%) (p=0.02). Strong PINCH stromal immunostaining predicted nodal metastasis: 19/26 (73%) OSCC cases with nodal metastasis had strong PINCH immunostaining compared to 9/31 (29%) cases without nodal metastasis (p=0.02). The PINCH expression in OSCC was more intense in stroma at the invasive edge than in intratumoural stroma. In conclusion, the up-regulation of PINCH protein in stroma may be involved in promoting invasion and metastasis in OSCC. PMID- 16273249 TI - Inhibition of cytotoxicity of cisplatin by cyclooxygenase-2 inhibitor nimesulide in head and neck cancer cell lines. AB - Although head and neck cancer is a common malignancy, investigations have not yet improved the poor prognosis of patients. Therefore, it is important to find new cancer treatment modalities. Recent studies showed that cyclooxygenase, especially its isoform cyclooxygenase-2, is involved in tumorigenesis, tumor growth and metastasis. Inhibition of this enzyme by cyclooxygenase inhibitors has been shown to be antiproliferative in numerous cancer cell lines. The aim of this study was to investigate if the selective cyclooxygenase-2 inhibitor nimesulide could enhance cytotoxicity of the standard chemotherapeutic agent cisplatin. Head and neck squamous cell cancer cells were incubated with nimesulide and/or cisplatin, and counted after 24, 48 and 72 h treatment. Visualization of apoptotic cells was done by immunohistochemistry. We demonstrated that nimesulide inhibits the cytotoxic effect of cisplatin in HNSCC cells. Therefore, COX-2 inhibitor nimesulide may not be a good partner for cisplatin in combination therapy for cancer. PMID- 16273251 TI - Disturbance of circadian gene expression in endometrial cancer: detection by real time quantitative RT-PCR. AB - Circadian genes control the daily changes of the circadian rhythms in a variety of physiological processes, which in turn regulate many functions in the human body. Disruption of circadian rhythms can have a profound influence on our well being. We established a set of PCR primers and fluorescent probes to analyze the mRNA levels of nine different circadian genes, and used immunohistochemical methods to study four important circadian proteins in 35 endometrial cancers and their paired non-cancerous tissues. Of these, 13 cases showed reduced expression in all nine circadian genes in the cancerous tissues relative to the paired non cancerous tissues; the remaining cases showed similar reduced expression in 4-8 of the genes analyzed. Conversely, 3 non-cancerous tissues showed reduced expression in all nine circadian genes in comparison with their respective adjacent cancerous tissues, whereas 6 other non-cancerous tissues showed reduced expression in 6-8 of the circadian genes. These results were also confirmed by immunohistochemical study. Expression of the circadian genes is perturbed in endometrial cancer. Based on these results, we suggest that different circadian rhythms occur in endometrial cancer and non-cancerous tissues. Our results may provide the molecular basis for chronotherapy of endometrial cancer. PMID- 16273250 TI - Cell-cycle-associated markers and clinical outcome in human epithelial cancers: a tissue microarray study. AB - The development and progression of epithelial cancers are the result of an imbalance in signals promoting and inhibiting cellular proliferation and apoptosis. The aim of this study is to evaluate the expression of cell-cycle and apoptosis regulators and correlate them with clinical outcome in the most frequent carcinomas, in order to establish common prognostic biomarkers independent of cancer origin. Using tissue microarrays (TMAs), we have analysed the immuno-expression of Ki-67, Bcl-2, Bax, cyclin D1, cyclin D3, CDK1, CDK2, CDK6, p16, p21, and p27 in a series of 205 carcinomas of the large bowel, breast, lung and prostate (80, 73, 37 and 15 cases, respectively). By univariate analysis, positivity for p27, p16 and Bcl-2 was associated with better overall survival (P<0.0135, P<0.0442 and P<0.0001, respectively). The risk of mortality was 2.3-fold greater in patients without Bcl-2 expression. TMA immunohistochemical analysis identified a subset of epithelial cancers with overlapping alterations in cell-cycle checkpoints, apoptosis regulators and tumour suppressor pathways. We found that in most common epithelial cancers, regardless of origin, Bcl-2 appears to be the key biological factor influencing clinical behaviour. PMID- 16273252 TI - First-line systemic treatment with gefitinib in stage IV non-small cell lung cancer. AB - Lung cancer has a high mortality rate and is often diagnosed in locally advanced or metastatic stages. A new therapeutic option for patients with non-small cell lung cancer (NSCLC) in these stages with progress or relapse after platinum-based chemotherapy exists in the inhibitors of the epidermal growth factor receptor (EGFR) tyrosine-kinase. EGFR tyrosine-kinase inhibitor treatment might also be an option for patients ineligible for surgery and conventional chemotherapy. We present a case of a 53-year-old woman who was diagnosed due to pain from multiple bone metastases of a lung adenocarcinoma. She refused cytotoxic chemotherapy, and we administered first-line systemic treatment with gefitinib subsequent to radiotherapy of metastatic bone disease. The patient responded well to gefitinib treatment and achieved a partial response after 3 weeks. No relevant side effects occurred, and the patient experienced an 8-month remission of disease. With a follow-up of 10 months, the patient is still alive. Retrospectively, we found a mutation of the EGF receptor in tumor cells of the patient, which is associated with sensitivity to gefitinib. EGFR tyrosine-kinase inhibitors (TKI) can be an alternative first-line systemic treatment option for selected patients with metastatic NSCLC. PMID- 16273253 TI - Combination hepatic arterial infusion therapy is effective for ocular melanoma metastasis to the liver. AB - Regional treatments including chemotherapy have been applied to patients with malignant melanoma metastasis to the liver in clinical trials, but with limited efficacy. To improve efficacy, combination therapy may be beneficial. We had a case of multiple liver metastasis from right ocular malignant melanoma. The primary melanoma was surgically resected, and combination hepatic arterial infusion (HAI) therapy using dacarbazine, nimustine, vincristine (DAV), and cisplatin was applied to the metastatic focus. After HAI, marked regression of the liver metastatic focus was observed. Unfortunately, the patient passed away due to involvement of thrombotic thrombocytopenic purpura 9 months after the initial consultation, however, aggravation of the liver metastatic focus was not observed. Combination HAI therapy may have a potent therapeutic effect on malignant melanoma metastasis to the liver, and our regimen using DAV and CDDP may be beneficial to the improvement of prognosis without serious side effects. PMID- 16273255 TI - Nicotine induces chromatin changes and c-Jun up-regulation in HL-60 leukemia cells. AB - Although nicotine has been implicated as a potential factor in the pathogenesis of human cancer, its mechanisms of action regarding cancer development remain largely unknown. HL-60 cells were used to investigate the effects of a short-term treatment with nicotine at concentrations found in the blood of smokers. The findings show that nicotine induces chromatin decondensation, histone H3 acetylation and up-regulation of the c-Jun transcription factor mRNA. This increase is inhibited by mecamylamine, a nicotinic receptor antagonist, suggesting that nicotine alters cellular function directly via nicotinic acetylcholine receptors and may then play a role in cell physiology and tumor promotion. PMID- 16273254 TI - Topotecan plus ifosfamide in patients with platinum refractory advanced/metastatic non-small cell lung cancer: a phase II trial. AB - A number of second line treatments have been proposed in patients with advanced pretreated non-small cell lung cancer (NSCLC). However, either single agents or two or three drug combinations achieved very poor results with no superiority of any combination over monotherapy. We have treated 42 patients (30 males) affected by advanced/metastatic NSCLC progressing during front line cisplatin-based chemotherapy with a combination of topotecan (1.2 mg/m2) plus ifosfamide (1200 mg/m2) for 3 consecutive days every 3 weeks. The median age was 63 years (range 43-76); cell types were: squamous carcinoma (n=17), adenocarcinoma (n=16), large cell carcinoma (n=3), broncho-alveolar carcinoma (n=2) and undifferentiated carcinoma (n=4). All patients were treated with a platinum containing chemotherapy: 39 patients with cisplatin, 2 patients with carboplatin and 1 patient with oxaliplatin, respectively. The ECOG PS was 0 in 8 patients (19%), 1 in 11 patients (26%), and 2 in 23 patients (55%). The median number of courses administered was 3 (range 1-8). Grade 3-4 neutropenia was the dose limiting toxicity, observed in 36% of patients. Moreover, grade 3-4 anemia and thrombocytopenia were observed in 17% and in 12% of patients, respectively. One PS 2 patient died of grade 4 hematological toxicity after the first cycle. No complete response was observed. Six (14.2%) subjects obtained a partial response (PR). In addition, 1 (2.4%) minimal response (MR) plus 14 (34%) stable diseases (SD) and 21 (51%) progressive diseases (PD) were observed. Median time to disease progression and median survival were 9 weeks (range 1-13) and 26 weeks (range 1 91+), respectively. The 1-year survival rate was 14%. Combination of topotecan and ifosfamide demonstrated antitumor activity in patients with relapsing or refractory NCSLC with a modest side effect profile and an overall disease control (PR + MR + SD) of 50.7%. Nevertheless, the still low response rate and the shortness of median survival indicates the need for more effective second line treatments in this disease. PMID- 16273256 TI - Decrease in size of azoxymethane induced colon carcinoma in F344 rats by 180-day fermented miso. AB - The present study was designed to investigate the effects of fermented miso (fermented soybean paste) on the induction of colon tumors by azoxymethane (AOM) in male F344 rats. A total of 91 rats, 6 weeks of age, were divided into 5 groups and given weekly subcutaneous injections of AOM (15 mg/kg body wt) for 3 weeks. The animals were placed on diets one week before the first AOM dose: commercial normal control MF diet or a diet containing 10% 2-year, 180-day fermented, or 3-4 day fermented miso. There were no differences in body and organ weights, and no aberrant crypt foci (ACF) among carcinogen-treated groups at week 25. The rates of tumor incidence were 45%, 85%, 75% and 60% with the 2-year, 180-day, and 3-4 day fermented miso and MF, respectively, and those for colon tumors were 34%, 55%, 60% and 55%, respectively. The size of well-differentiated adenocarcinomas and total (well differentiated and signet ring cell) adenocarcinomas in the 180 day fermented miso group was significantly smaller than that in the 2-year fermented miso and MF+AOM groups. Nuclear staining of beta-catenin in colon tumors was increased for the 3-4-day fermented miso compared to the 180-day fermented miso. Cdx2 staining tendency was decreased in colon tumors and adenocarcinomas compared to normal mucosa and ACF, which stained in 100% of cases. In addition, the PCNA index was significantly reduced in the 180-day group compared with those groups receiving the 3-4-day fermented miso and MF diet. The germinal region was also decreased. The present results indicate that dietary supplementation with 180-day fermented dietary miso could act as a chemopreventive agent for colon carcinogenesis. PMID- 16273257 TI - Vascular endothelial growth factor and soluble FLT-1 receptor interactions and biological implications. AB - Vascular endothelial growth factor (VEGF), binding to an appropriate receptor like FLT, is the main mitogen for endothelial cells and a strong inducer of angiogenesis. A soluble form of VEGF receptor, sFLT-1, specifically binds VEGF and inhibits its activity. The following expression plasmids were used in the experiments: pVEGF plasmid encoding VEGF165, pFGF-2 encoding FGF-2 and psFLT-1 plasmid encoding the soluble form of VEGF receptor, sFLT-1. The interaction between VEGF and sFLT-1 was evaluated using a migration test and ERK1/2 activity utilizing mouse sarcoma cells (L-1). Implication of the VEGF/sFLT-1 action was also visualized using in vivo angiogenesis assay. The conditioned medium (CM) from L-1 phVEGF-165 transfectants stimulated L-1 cell migration more than medium from non-transfected L-1 cells. Media collected from phVEGF-165 transfectants or original L-1 cells only slightly stimulated the migration of cells transfected with psFLT-1. The L-1 cells also showed intensive phospho-ERK1/2 activity when treated with the CM from VEGF transfectants. In vivo tests showed that sFLT-1 effectively suppressed VEGF-mediated angiogenesis without affecting FGF-2-driven angiogenesis. To summarize, this study documented that sFLT-1 released from transfected cells might inhibit cell functions induced by VEGF, but not by FGF. The results obtained from in vivo angiogenesis tests also confirm the antiangiogenic potency of cloned sFLT-1, which can be useful for planning cancer experimental therapy studies. PMID- 16273258 TI - Improved survival using multi-modality therapy in patients with lung metastases from colorectal cancer: a preliminary study. AB - This retrospective study was conducted to assess the safety, efficacy, and long term results of multi-modality therapy including radio-frequency thermal ablation (RFA) and radiotherapy as an additional cytoreductive method for eliciting the marked effects of chemotherapy in treating unresectable lung metastases from colorectal cancer. Total of 21 patients with lung metastasis from colorectal cancer were included. They were treated with modified pharmacokinetic modulating chemotherapy (PMC). Eleven were also treated with RFA and/or radiotherapy (multi modality group), and 10 were treated with chemotherapy alone (chemotherapy group). Characteristics and survival of patients in the multi-modality group were compared with those of the chemotherapy group. The median survival of all patients was 38.6 months after the initial PMC. The cumulative 3-year survival rate of patients in the multi-modality group was 87.5% compared with 33.3% in the chemotherapy group (p=0.0041). The course of multi-modality therapy was uneventful except for pneumothorax in those who received RFA. Although pneumothorax developed in 4 of 11 patients (36.4%) treated with RFA, all were able to receive chemotherapy within 2 weeks after RFA. In conclusion, multi modality therapy combined with modified PMC, radiation and RFA is a feasible choice of treatment associated with reasonable morbidity and mortality in patients with inoperable lung metastases from colorectal cancer. PMID- 16273259 TI - Complex formations involving both SP-1 and SP-3 at the transcriptional regulatory sequence correlate with the activation of the Keratin 14 gene in human oral squamous cell carcinoma cells. AB - We have previously reported that significantly higher levels of Keratin 14 (Ker 14) was observed in oral squamous cell carcinoma (OSCC) and severely dysplastic tissues, whereas this expression was reversed in hyperplasia and in mild to moderate dysplasia. In this study, the mechanism of Keratin 14 activation in oral squamous cell carcinoma (OSCC) cell lines (HSC-2, HSC-3 and Ca9-22) was investigated. Reporter analysis demonstrated that an upstream region (-1759/ 1629) accounted for efficient promoter activity. Furthermore, electromobility sift and supershift assay demonstrated that interactions of the SP-1/SP-3 complex at the elements resided in -1737/-1702 and -1680/-1652 and may be essential for this activation in OSCC cells. PMID- 16273260 TI - WNT/PCP signaling pathway and human cancer (review). AB - WNT/planar cell polarity (PCP) signaling pathway controls tissue polarity and cell movement through the activation of RHOA, c-Jun N-terminal kinase (JNK), and nemo-like kinase (NLK) signaling cascades. PCP is induced in Drosophila by the asymmetrical localization of Frizzled-Dishevelled-Diego-Starry night (Flamingo) complex and Van Gogh (Strabismus)-Prickle complex. Here, WNT/PCP signaling pathway implicated in human carcinogenesis is reviewed. Human WNT5A, WNT5B, and WNT11 are representative non-canonical WNTs transducing PCP signals through FZD3 or FZD6 receptors, and ROR1, ROR2 or PTK7 co-receptors. Human VANGL1, VANGL2 (Van Gogh homologs), CELSR1, CELSR2, CELSR3 (Starry night homologs), DVL1, DVL2, DVL3 (Dishevelled homologs), PRICKLE1, PRICKLE2 (Prickle homologs), and ANKRD6 (Diego homolog) are core PCP signaling molecules. MAGI3 assembles FZD, VANGL, PTEN, and adhesion molecules. Dishevelled-dependent WNT/PCP signals are transduced to the RHOA signaling cascade through Formin homology proteins DAAM1 and DAAM2, and to the JNK signaling cascade through MAPKKKs and MAPKK4/7. Dishevelled-independent WNT/ PCP signals are transduced to the NLK signaling cascade through MAP3K7 (TAK1). ANKRD6, NKD1 and NKD2 induce class switch from the WNT/GSK3beta signaling pathway to the WNT/PCP signaling pathway. WNT5A is up-regulated in various types of human cancer, such as gastric cancer, lung cancer, and melanoma. FZD3/FZD6 receptor and ROR2 co-receptor transduce WNT5A signal in gastric cancer. Aberrant activation of WNT/PCP signaling pathway in human cancer leads to more malignant phenotypes, such as abnormal tissue polarity, invasion, and metastasis. cDNA-PCR, microarray or ELISA reflecting aberrant activation of WNT/PCP signaling pathway could be developed as novel cancer prognostics. Single nucleotide polymorphism (SNP) and copy number polymorphism (CNP) of WNT/PCP signaling molecules mentioned above are suitable for use in screening of cancer predisposition, especially for gastric cancer. Antibody, RNAi, or small molecule compounds to regulate the function of WNT/PCP signaling molecules mentioned above are good candidates for development as novel cancer therapeutics. PMID- 16273261 TI - Thiamin deficiency: a possible major cause of some tumors? (review). AB - Based solely on clinical clues from a malnourished population, thiamin alone was intentionally and successfully injected to human cases with some tumors or masses. Two cases of submandibular gland cyst and 13 out of 15 cases of Baker's cyst were cured without recurrence for several decades. In a case with pathology confirmed osteosarcoma, subcutaneous perfusion of thiamin HCl 300 once only reduced its circumference from 30 to 20 cm, equivalent to a reduction of 50-75% in volume, within 2 days. Current concepts on the role of thiamin in carcinogenesis are controversial. Some authors claimed that thiamin supported high rate of tumor cell survival, proliferation and chemotherapy resistance and suggested anti-thiamin therapy for cancer. On the other hand, some investigators have reported evidence of prevention of several varieties of cancers by dietary thiamin. A limited number of animal studies revealed evident relationship between thiamin deficiency and cancer development. Therefore, further study on the mechanism switching thiamin between cancer supporter and suppressor is needed. PMID- 16273262 TI - The role of serotonin in tumour growth (review). AB - Serotonin (5-hydroxytryptamine; 5HT) a monoamine neurotransmitter mediates a wide range of physiological actions in the human body. For example 5HT is implicated in psychiatric and neurological disorders and also plays a fundamental role in tumour growth, differentiation and gene expression. 5HT acts as a growth factor for several types of tumoural and non-tumoural cells. This review considers the role of 5HT and its receptors in the human body with particular reference to carcinogenesis. We conclude that 5HT causes growth proliferation and 5HT antagonists cause growth inhibition in a variety of tumour cells (e.g. prostate carcinoma, lung carcinoma and colonic carcinoma). Therefore, further studies should look into the potential use of 5HT antagonists in the treatment of cancer. PMID- 16273263 TI - Artesunate in the treatment of metastatic uveal melanoma--first experiences. AB - Artesunate (ART) is a derivative of artemisinin, the active principle of the Chinese herb Artemisia annua L. Artesunate is approved for the treatment of multidrug-resistant malaria and has an excellent safety profile. It has been shown that Artesunate, apart from its anti-malarial activity, has cytotoxic effects on a number of human cancer cell lines, including leukemia, colon cancer and melanoma. We report on the first long-term treatment of two cancer patients with ART in combination with standard chemotherapy. These patients with metastatic uveal melanoma were treated on a compassionate-use basis, after standard chemotherapy alone was ineffective in stopping tumor growth. The therapy regimen was well tolerated with no additional side effects other than those caused by standard chemotherapy alone. One patient experienced a temporary response after the addition of ART to Fotemustine while the disease was progressing under therapy with Fotemustine alone. The second patient first experienced a stabilization of the disease after the addition of ART to Dacarbazine, followed by objective regressions of splenic and lung metastases. This patient is still alive 47 months after first diagnosis of stage IV uveal melanoma, a situation with a median survival of 2-5 months. Despite the small number of treated patients, ART might be a promising adjuvant drug for the treatment of melanoma and possibly other tumors in combination with standard chemotherapy. Its good tolerability and lack of serious side effects will facilitate prospective randomized trials in the near future. PMID- 16273264 TI - Demographic and psychosocial factors associated with risk perception for breast cancer. AB - The objective of this study was to investigate, in a female population-based sample, specific demographic and psychosocial predictors of perception of risk for female breast cancer. The study was a cross-sectional survey of 7135 healthy women ranging from 15 to 85 years. A questionnaire assessed the perceived risk for breast cancer in 6 German centers. Specified awareness was personal knowledge in relation to breast cancer incidence, risk factors, perception, and level of concern. An awareness score was defined and related to values of several risk factors. Generally, 78.1% of women were aware of breast cancer. However, only one third (31.2%) of women estimated the incidence of breast cancer correctly; 52% recognized age, 33.9% recognized hormonal contraceptives, and 34.1% recognized hormonal replacement therapy as factors of breast cancer risk. The most frequent specified sources of information were gynecologists (48.4%). The awareness score of women informed by gynecologists was significantly better compared to women informed by other healthcare professionals. However, a statistically significant improvement of the awareness score was reached for women with a higher educational level, information seeking by TV, radio, printed sources, medical books and healthcare flyers. Recommendations for improvement of cancer prevention programs include targeting comprehension regarding the lifetime risk of breast cancer, age as a risk factor, survival from breast cancer and hormonal factors. According to these results, there is an urgent need to separately address the perceptions of women, depending on age, social status and educational level. PMID- 16273265 TI - Down-regulation of PEDF expression by ribozyme transgene in endothelial and lung cancer cells and its impact on angiogenesis in vitro. AB - Pigmental epithelial-derived factor (PEDF) is an important anti-angiogenesis factor. It has many different functions in ocular cells. The inhibition on angiogenesis by PEDF is associated with endothelial cell apoptosis. Until now, there have been few reports on the PEDF's role in cancer cell progression. This study examined the effect of a ribozyme transgene, designed to inhibit human PEDF expression, and its impact on in vitro lung cancer, endothelial cell growth and the angiogenesis forming process, in vitro. A transgene encoding ribozymes to specially target human PEDF was constructed using pEF6/v5-his vector. The human lung cancer cell line, A549, and endothelial cells, HECV, were transfected with PEDF ribozyme. MTT assay was used to analyse cell growth alternation after the PEDF gene was knocked out. An in vitro endothelial tubule formation assay was employed to analyse the microtubule forming change after the PEDF gene was knocked out in HECV cells. The PEDF message and protein were successfully removed with the PEDF ribozyme as shown by reverse transcription-PCR and Western blotting, respectively. The growth of both A549 and HECV cells were significantly accelerated when the PEDF gene was knocked out. The HECV cell ability to form microtubules increased following PEDF knockout. Furthermore, HECV significantly enhanced the ability to form microtubules when co-cultured with A549 cells whose PEDF expression was lost by way of the ribozyme transgene. Ribozyme transgenes targeting PEDF in lung cancer and endothelial cells can reduce the growth of these cells. Expression of PEDF in both cancer cells and endothelial cells are correlated with the angiogenic process' paracrine and autocrine pattern. PMID- 16273266 TI - Aberrant methylation: common in thymic carcinomas, rare in thymomas. AB - Thymic carcinoma, which is a rare epithelial neoplasm of the thymus gland, is different from thymoma in its clinical and pathological features. To clarify the mechanism underlying the aggressive behavior of thymic carcinoma, we examined the clinicopathologic features, aberrant methylation patterns of the tumor suppressor genes, and epidermal growth factor receptor (EGFRs) mutation in both thymic carcinomas and thymomas. Clinical data of 11 thymic cancers and 13 thymomas were reviewed. Resected samples of 5 thymic cancers and 6 thymomas selected from 24 cases were used for methylation and mutation studies. Positive tumor markers were more frequent in thymic cancers than in thymomas (p=0.0233), and the methylation index, which reflects the overall methylation pattern, was significantly higher in thymic carcinomas (p=0.0053). No tumors showed a mutation of EGFR, KRAS, and HER2. Thymic carcinoma is distinct from thymoma not only with respect to clinicopathological features, but also aberrant methylation patterns of the tumor suppressor genes. PMID- 16273267 TI - Preventive effect of the soluble tumor-associated antigens on DMBA induced tumorigenesis in C3H/He mice. AB - In our previous studies, we showed that soluble tumor-associated antigens (sTAA) of 66 kDa and 51 kDa have distinct tumor-preventive effects on chemically induced mammary cancer in rats and are able to repair the damage caused by tumorigenesis in its early stages. In the present study, we investigated whether these proteins can prevent the development of chemically induced tumors in mice. The study was performed on C3H/He mice which have the ability to develop many spontaneous tumors with age. Forty-four, 6-week-old mice were exposed twice at a 2-week interval to the carcinogen 9,10-dimethyl-1,2-benz(alpha)anthracene (DMBA), at a dose of 2 mg/mouse administered intragastrically. Two months later, the mice were divided into two groups. One group received sterile saline twice a week at a dose of 0.2 ml/mouse, intraperitoneally (i.p.). The other group received sTAA twice a week at a dose of about 10 microl in 0.2 ml of sterile saline/mouse, i.p. Periodically, all mice were checked for the presence of tumors. The experiment was terminated at week 35. Vaccination with sTAA increased the time of involvement of mice in the experiment, prevented the tumorigenic effect of DMBA, and inhibited further development of existing tumors. PMID- 16273268 TI - Radical radiotherapy alone for glomus jugulare and tympanicum tumours. AB - The management of glomus jugulare and tympanicum tumours is controversial due the long natural history and morbidity associated with intervention. We report the experience of radical radiotherapy (RT) alone for these tumours. Between 1965 and 1987, 49 patients received RT. Median age at presentation was 55 years (range, 23 82). Common presentations were deafness in 27 patients, tinnitus in 25 and cranial nerve palsies in 18. RT was given as a 2D simulator-planned wedge pair in the majority with a median dose of 45 Gy (range, 37.5-50.0) in 15 or 16 fractions over 21 days (range, 20-26). Median follow-up was 7.4 years (range, 2.0-23.4). At 6 months post-RT, complete clinical response was seen in 38 patients, partial response in 4, no response in 1 and no data were available for 6. At both 5 and 10 years, 92% of patients were recurrence-free and cancer-specific survival was 96%. There were no reports of radionecrosis. Although tumour eradication is not the aim, RT can achieve good local control, survival and symptom relief without the significant morbidity that can be associated with radical surgery. Therefore, RT alone has a significant role in the management of these tumours. PMID- 16273269 TI - The proteasome inhibitor MG132 induces apoptosis in human pancreatic cancer cells. AB - The ubiquitin-proteasome system plays a critical role in the regulation of programmed cell death. Proteasome inhibitors induce apoptosis in various cancer cells and have antitumor effects in murine tumor models. In the present study, we investigated whether the cell-permeable proteasome inhibitor MG132 (carbobenzoxyl L-leucyl-L-leucyl-L-leucinal) reduced the growth of a human pancreatic cancer cell line through induction of apoptosis in vitro. The effects of MG132 (0.125 1.000 microM) on the growth of the human pancreatic cancer cell line BxPC-3 were analyzed by cell count and MTT assay. Apoptosis was determined by FACS analysis after annexin V and propidium iodide staining and the enrichment of intracellular nucleosomes. The proteasome inhibitor MG132 decreased cell growth of the human pancreatic cancer cell line BxPC-3 in a dose- and time-dependent manner. This effect was at least in part mediated by the induction of apoptosis. A combination therapy with standard cytotoxic agents and proteasome inhibitors could potentially be a novel therapeutic strategy in treatment of pancreatic cancer. PMID- 16273270 TI - A novel region of deletion on 13q33-q34 in esophageal squamous cell carcinoma. AB - Chromosome 13 presents frequent allelic loss in esophageal squamous cell carcinomas (ESCC). However, no ESCC suppressor gene has been identified from this chromosome. To define common deletion regions that possibly contain the ESCC suppressor gene(s), we performed a mapping of allelic loss in 50 esophageal squamous cell carcinomas using a panel of 25 microsatellite markers on chromosome 13q21-qter, which has rarely been studied for allelic loss. Loss of heterozygosity (LOH) with high frequencies (> or = 50%) was observed at markers D13S1494, D13S1323, D13S248, D13S1315, D13S285, and D13S1295, in which the peak LOH (69.2%) was at locus D13S248. Seven cases presented LOH at three consecutive markers D13S248, D13S1315 and D13S285, 4 of which also displayed LOH at another adjacent marker D13S1295. This overlapping region of deletion covers an interval of 6.36 Mb at 13q33.1-q34, whose deletion has not previously been reported in ESCC. Tumors of grade II showed significantly more frequent LOH at D13S248 than those of grade I. A significantly higher frequency of allelic loss at D13S152 was also found in tumors with lymph node metastasis compared to those without lymph node metastasis. The present study defined a novel region of allelic loss in 13q33-q34. LOH at D13S248 and D13S152 are associated with higher tumor grade and metastasis, respectively. PMID- 16273271 TI - Thoracic radiation therapy and concomitant low-dose daily paclitaxel in non-small cell lung cancer: a phase I study. AB - The primary objective of the current study was to determine the maximum tolerated dose of paclitaxel (PTX) that could be added daily to radiation therapy (RT) in patients with non-small cell lung cancer (NSCLC). The secondary objective was to achieve a plasma concentration of PTX estimated to exert a radiosensitizing effect. Eighteen patients with locally advanced NSCLC were treated. 60 Gy of RT were given in 2 Gy fractions, 5 days per week. A daily dose of PTX was delivered by 4-h i.v. infusion before RT. The initial dose level of PTX was 9 mg/m2/day, escalated by 1 mg at each additional patient triplet. Two out of 6 patients experienced acute dose limiting toxicity (DLT) at the 12 mg/m2/day PTX dose level. PTX continuously greater than 10 nM, the estimated radiosensitizing condition, was achieved at the PTX dose level of 12 mg/m2/day. PTX at doses up to 11 mg/m2/day may be safely added to a conventional conformal RT course. Both DLT and the estimated radiosensitizing plasma exposure to PTX were encountered at the 12 mg/m(2)/day dose level. PMID- 16273273 TI - Analysis of tumor progression by transcriptional profiling of mouse MK16 cell lines transformed with human papillomavirus type 16 E6 and E7 oncogenes and activated H-ras. AB - A better understanding of the molecular basis of tumor progression and invasion is needed to improve therapy for malignant tumors. Recently, we established a mouse metastatic MK16 model by transduction of secondary kidney cells with human papillomavirus type 16 (HPV16) E6 and E7 oncogenes and human H-ras activated by G12V mutation. In this study, we extended the model to MK16 cell lines derived from lung metastases and compared the oncogenicity of seven cell lines successively isolated from primary tumors or metastases. By observing the formation and growth of subcutaneous tumors and generation of lung metastasis, we showed a gradual increase in oncogenicity of MK16 cell lines. Interestingly, we demonstrated metastatic potential of MK16/A cells with low oncogenic potential in primary tumor development. To detect changes in gene expression associated with increasing oncogenicity of MK16 cell lines, we performed transcriptional profiling with the Atlas Plastic Mouse 5K microarray. We found that a substantial proportion of up-regulated genes encoded ribosomal proteins. Among the down regulated genes, the highest number (n=10) belonged to a group coding for transcription factors. Expression of two of these, Pou3f2 and Gtl3, was reduced both in cells derived from primary tumors and those isolated from metastases. Furthermore, microarray hybridization suggested that the down-regulation of cyclin-dependent kinase inhibitors p16(Ink4a) and p57(Kip2) and up-regulation of A6 and A10 members of the S100 protein family might play a role in the increase of MK16 oncogenicity. PMID- 16273272 TI - Expression and prognostic value of EGFR in invasive breast cancer. AB - Epidermal growth factor receptor (EGFR) is a membrane receptor expressed in a variety of solid human cancers and directly related with poor prognosis. The objective of this work was to evaluate the EGFR content in breast carcinomas, its possible relationship with different clinical-pathological parameters, and its potential prognostic significance and predictive value. EGFR levels were examined by radioligand binding assays in 846 patients with invasive breast cancer. The median follow-up period was 50 months. There was a wide variability of EGFR levels among the studied tumors (0.01-403 fmol/mg protein). Statistical analysis showed that EGFR levels were significantly higher in younger patients (p=0.0001). EGFR were also notably higher in ER-negative or PgR-negative tumors than in ER positive (p=0.0001) or PgR-positive tumors (p=0.001). In addition, the presence of high intratumoral EGFR levels (cut-off: 6 fmol/mg protein) was associated with both shorter relapse-free survival (p=0.04) and overall survival (p=0.01) in the group of patients as a whole, as well as with overall survival in the subgroup of patients without any type of systemic adjuvant treatment (p=0.02). However, EGFR levels did not achieve significance as independent prognostic factor in the multivariate analysis. There is a wide variability of intratumoral EGFR levels in breast carcinomas, and these protein levels correlated positively with a poor prognosis in the t univariate analysis. However, further studies are necessary in order to assess the possible clinical value of EGFR in combination with other essential components of the EGFR family network. PMID- 16273274 TI - Thimerosal induces neuronal cell apoptosis by causing cytochrome c and apoptosis inducing factor release from mitochondria. AB - There is a worldwide increasing concern over the neurological risks of thimerosal (ethylmercury thiosalicylate) which is an organic mercury compound that is commonly used as an antimicrobial preservative. In this study, we show that thimerosal, at nanomolar concentrations, induces neuronal cell death through the mitochondrial pathway. Thimerosal, in a concentration- and time-dependent manner, decreased cell viability as assessed by calcein-ethidium staining and caused apoptosis detected by Hoechst 33258 dye. Thimerosal-induced apoptosis was associated with depolarization of mitochondrial membrane, generation of reactive oxygen species, and release of cytochrome c and apoptosis-inducing factor (AIF) from mitochondria to cytosol. Although thimerosal did not affect cellular expression of Bax at the protein level, we observed translocation of Bax from cytosol to mitochondria. Finally, caspase-9 and caspase-3 were activated in the absence of caspase-8 activation. Our data suggest that thimerosal causes apoptosis in neuroblastoma cells by changing the mitochondrial microenvironment. PMID- 16273275 TI - Time-dependent RNA degradation affecting cDNA array quality in spontaneous canine tumours sampled using standard surgical procedures. AB - Heterogeneous gene expression in tumours and the degradation of RNA when sampling under non-RNAse-free conditions may limit the potential benefit of cDNA array studies. This study examines changes in the integrity of RNA by means of RNA gel electrophoresis at various post-operative intervals on canine mammary tumours (n=10) and malignant lymphoma (n=1). The tumours were cut into pieces (3-5 mm diameter, approximately 50 mg) and kept in tubes without RNAse-free buffer at room temperature. No special precautions were taken to avoid the influences of Rnase; rather, normal surgical procedures were used. We found that total RNA of the mammary tumours started to degrade within 30 min of the operation, and the rate of degradation increased up to 4 h, which was the last time point included in this study. RNA in the lymphoma tumours degraded more rapidly, and was completely degraded at 30 min post-operation. The degradation of mRNA in the mammary tumours, as studied by human cDNA arrays, was heterogeneous, i.e. some mRNA degraded completely, some only partially. This indicates that the mRNA degradation rate varied depending on the type of mRNA. However, since we found that gene expression differs depending on the part of the mammary tumour examined, one cannot exclude that the variation in the mRNA degradation rate may simply reflect heterogeneous gene expression within the tumour. We conclude that RNA integrity is unaffected immediately after sampling under non-RNAse-free conditions; however, the tumour sample should be preserved under RNAse-free conditions within 15 min to avoid RNA degradation. This is a much shorter time interval than previously reported in other similar studies; however, these studies generally treated normal tissue, under which 3-5 h non-RNAse-free conditions have been found not to affect RNA quality. PMID- 16273276 TI - Activation of ectopic Oct-4 and Rex-1 promoters in human amniotic fluid cells. AB - Recently, amniotic fluid was suggested as a new source for stem-cell research and tissue engineering approaches. In order to enable isolation of stem cells and establishment of lines of such cells with an undifferentiated phenotype we have introduced green fluorescent protein regulated by the promoters of the stem cell specific genes, Oct-4 or Rex-1, into human amniotic fluid cells. For the introduction of DNA into human amniotic fluid cells, we have optimized a specific transfection protocol. We found that human amniotic fluid contains cell populations which are able to activate these promoters. These undifferentiated cells expressing green fluorescent protein can be analysed on a flow cytometer. In addition, we have introduced a plasmid harboring a neomycin-resistance gene under the control of the Oct-4 promoter. G418 selection allowed the isolation of undifferentiated stem cells expressing Oct-4 protein out of human amniotic fluid samples. Our findings confirm the existence of stem cells within amniotic fluid. In addition, the ability to transfect human amniotic fluid cells and to isolate stem-cell marker-positive cells will provide the means to study and manipulate these cells for the purpose of basic and applied research. PMID- 16273277 TI - Anti-angiogenic effects of Hypericin-photodynamic therapy in combination with Celebrex in the treatment of human nasopharyngeal carcinoma. AB - Photodynamic therapy (PDT) is being investigated as an alternative treatment modality in cancer treatment. It has been shown to induce tumor hypoxia and upregulation of cyclooxygenase-2 (COX-2) and vascular endothelial growth factor (VEGF). The objective of this study was to improve in vivo tumor growth control of nasopharyngeal carcinoma (NPC), treated at a subcurative dosage by using a combination of Hypericin-PDT and COX-2 inhibitor, Celebrex (CX). The effect of an initial CX dose at 6- and 24-h post-PDT was investigated simultaneously. It was observed that hypoxic NPC/CNE2 cells upregulate both COX-2 and VEGF A genes in vitro. In vivo studies, down-regulation of COX-2 and hypoxia inducible factor 1alpha (HIF-1alpha) genes at 24-h post-PDT and bulk tumor ablation at 48-h post PDT was observed. However, 24-28 days later regrowth was observed. In a combination treatment, 1st CX dose at 6-h post-PDT had the highest tumor control in which tumors were 1,400 pg/ml and >25, respectively. The resulting figures for sensitivity, specificity, and positive and negative predictive values were 78%, 93%, 78% and 93%, respectively. The achieved accuracy (89%) was significantly higher than that for the 14-3-3 test (67%; P=0.023). The high predictive values achieved by the combined tau test suggest that it could be used in conjunction with other diagnostic methods for accurate intra vitam diagnosis of CJD. PMID- 16273299 TI - HFE gene mutations in Polish patients with disturbances of iron metabolism: an initial assessment. AB - Hereditary hemochromatosis is one of the most frequent genetic disorders in Europeans, but its prevalence in Poland is still unknown. The aim of the study was an initial assessment of the prevalence of C282Y and H62D HFE gene mutations and their influence on the course of chronic hepatitis C. Forty-one patients were admitted to the Department of Infectious Diseases, Medical University of Gdansk in 2000-2004 because of chronic liver diseases with accompanying disturbances in iron metabolism. Genetic tests for the C282Y and H63D mutations were performed by PCR and restriction fragment length polymorphism (PCR-RFLP) analysis. The HFE gene mutations were confirmed in 24 of 41 (59%) cases with symptoms of chronic liver disease and iron overload, significantly more frequently in HCV-negative patients (12/14 vs. 12/27; chi2=8.28; p=0.05). The C282Y and H63D HFE gene mutations were detected in 16 of 41 (39%) and 9 of 41 (22%) cases, respectively. HCV-negative patients were C282Y carriers significantly more frequently than HCV positive patients [9/14 vs. 2/27 C282Y homozygotes; 2/14 vs. 3/27 C282Y heterozygotes (p<0.0001)]. The carrier state of the H63D HFE gene mutation was not significantly more frequent in HCV-positive than HCV-negative patients. HCV infection seems to be a negative predictive marker of HFE gene mutations in patients with iron overload. The relationship of H63D HFE gene mutations with chronic hepatitis C and the possible influence of HCV infection on iron metabolism needs further analysis. PMID- 16273301 TI - Mutation screening of three candidate genes, ELOVL5, SMAP1 and GLULD1 in autosomal recessive retinitis pigmentosa. AB - Retinitis pigmentosa (RP) is the most common form of retinal dystrophy. It is featured by a great clinical and genetic heterogeneity. Different patterns of inheritance exist, such as autosomal dominant and recessive, X-linked and digenic. RP25, a locus for autosomal recessive retinitis pigmentosa (arRP), the most frequently inherited form of RP, was mapped to chromosome 6q between D6S257 and D6S1644 microsatellite markers. ELOVL5, SMAP1 and GLULD1 were selected on the basis of their location, tissue expression and/or function. ELOVL5 is implicated in the elongation of long chain fatty acids, including docosahexanoic acid (DHA), which constitutes 50% of the fatty acids of the outer segment of the photoreceptor. SMAP1 (stromal membrane associated protein 1) was found to be located within RP25 locus and is expressed in retina. GLULD1, glutamate-ammonia ligase (glutamine synthase) domain containing 1, plays a key role in the uptake and metabolism of glutamate in the retina. The absence of pathogenic mutations after molecular analysis argues against the implication of ELOVL5, SMAP1 and GLULD1 in the development of RP25 phenotype. Nevertheless, we could not rule them out as good candidates for other retinal degeneration mapping to the same chromosomal region. PMID- 16273300 TI - Antiproliferative and apoptosis-inducing activity of Brucea javanica extract on human carcinoma cells. AB - We have recently demonstrated the antiproliferative and apoptotic activities of herbal traditional Chinese medicines, including the analomous fruit extract of Gleditsia sinensis, the fresh juice of Scutellaria barbata and the warmed water extract of Radix Sophorae Tonkinensis on a series of human carcinoma cells. Here, we further report the potential anti-cancer activity of the warmed water extract of Brucea javanica (BJE). Four cancer cell lines, including A549 non-small cell lung cancer, Hep3B hepatocellular carcinoma, MDA-MB231 breast cancer and SLMT-1 oesophageal squamous cell carcinoma, were incubated with BJE and strong apoptotic induction was observed under inverted microscopic investigation for all of the four cell lines tested. Using the MDA-MB231 breast cancer cell line as an experimental model, additional analyses supported the hypothesis that the mitochondrial membrane potential depolarization pathway was induced by BJE. The APO-1/Fas receptor death induction pathway was not activated under the influence of BJE, as studied by staining with Fas ligand and Fas receptor specific antibodies. Accordingly, only weak activation of caspase 8 was observed upon BJE treatment. On the other hand, caspase 3 activity was stimulated up to five-fold in BJE-treated cells compared to untreated controls. Oligonucleosomal DNA fragmentation formation was detected by labelling the nucleic acid ladders with TdT-mediated dUTP nick end labelling. Collectively, BJE-induced cancer cell death proceeds through a mitochondrial dependent pathway associated with caspase 3 activation. PMID- 16273302 TI - Aberrant expression of FGFRL1, a novel FGF receptor, in ovarian tumors. AB - FGFRL1 is a novel member of the FGF receptor family. It is expressed at very low levels in a great variety of cell lines and at relatively high levels in SW1353 chondrosarcoma cells, MG63 osteosarcoma cells and A204 rhabdomyosarcoma cells. Screening of 241 different human tumors with the help of a cancer profiling array suggested major alterations in the relative expression of FGFRL1 in ovarian tumors. Five distinct ovary tumors were therefore analyzed by quantitative and competitive PCR. Several tumors were found to exhibit a significant decrease in the expression of FGFRL1 in the tumor tissue relative to the matched control tissue. One ovarian tumor showed a 25-fold increase in the relative expression. Since FGFRL1 appears to be involved in the control of cell proliferation and differentiation, its aberrant expression might contribute to the development and progression of ovarian tumors. PMID- 16273303 TI - Mutational risk in highly repetitive exon ORF15 of the RPGR multidisease gene is not associated with haplotype background. AB - Exon ORF15 is an alternative exon in the retinitis pigmentosa GTPase regulator (RPGR) gene containing a highly repetitive, purine-rich internal region. It constitutes a mutational hot spot giving rise to a group of heterogeneous X linked retinal disorders. We sought to determine whether non-pathogenic substitutions and sequence length variations in the repetitive sequence have an influence on the risk of pathogenic exon ORF15 mutations. The type and distribution of exon ORF15 polymorphisms were assessed by genotyping 107 healthy German males using standard procedures. Polymorphisms were grouped into haplotypes and their frequencies determined in normal controls and previously analyzed patients with X-linked retinitis pigmentosa (XLRP). In the control group we identified 6 complex variants of the most common, ancestral exon ORF15 allele corresponding to the GenBank reference sequence (accession no. AF286472). Exon ORF15 mutations in XLRP patients were associated with the ancestral allele in 75% of affected cases. Four of the most recent founder haplotypes termed H3, H4, H6 and H7 were not identified in the patient samples. Our analysis and review of polymorphism data from the published literature suggests the presence of common exon ORF15 haplotypes in the European population. While the mutational risk in the RPGR gene appears not to be altered by the haplotype background, exon ORF15 haplotype analysis may be useful for tracing the evolutionary history of RP3 associated diseases. PMID- 16273304 TI - Conditionally replicative adenovirus vector carrying TRAIL gene for enhanced oncolysis of human hepatocellular carcinoma. AB - Conditionally replicative adenovirus (CRAd) mediated tumor specific gene therapy based on transcriptional control is considered a new direction for the treatment of cancer. Our previous studies showed that an HS4 insulator increased the alpha fetoprotein (AFP) promoter-driven expression in the context of an adenovirus (Ad) vector, while retaining the highly specific gene expression in hepatoma cells in vitro and in vivo. In this study, we constructed two HS4-AFP promoter based CRAd vectors (Ad.HS4.AFP.E1a/TRAIL and Ad.HS4.AFP.E1a) with and without the expression cassette of TNF-related apoptosis-inducing ligand (TRAIL). The TRAIL-expressing virus vector, Ad.HS4.AFP.E1a/TRAIL, exhibited more obvious oncolytic effect than Ad.HS4.AFP.E1a in both high-AFP-producing HCC cell lines (Hep3B and HUH7) and a low-AFP-producing HCC cell line (PLC/PRF/5) examined, indicating endogenous TRAIL over-expression increased CRAd potency. The enhanced hepatoma cell death was mainly mediated through apoptotic mechanism, as evidenced by the activation of caspase-3, binding of annexin V and inhibition by caspase inhibitor z-vad-fmk. In s.c. xenograft of low-AFP-producing PLC/PRF/5 hepatoma model, the administration of Ad.HS4.AFP.E1a/TRAIL resulted in a more potent oncolytic effect compared with the same dose of Ad.HS4.AFP.E1a 28 days after virus exposure. This study demonstrated that the TRAIL in the context of a CRAd vector was able to increase the oncolytic activity in low-AFP-producing HCC cells in vitro and in vivo. Considering that oncolytic viruses destroy tumor cells expressing low levels of the tumor marker is a clinical concern, TRAIL might be a useful tool to improve the efficacy of these vectors. PMID- 16273305 TI - Is the chimpanzee, Pan troglodytes, an endangered species? It depends on what "endangered" means. AB - I review information on the status in the wild of the robust chimpanzee, Pan troglodytes, and consider whether this evidence is consistent with the designation of P. troglodytes as Endangered in the International Union for the Conservation of Nature and Natural Resources (IUCN) Red List, and with public statements to the effect that great apes as a whole will be extinct within a few decades. Chimpanzees remain widespread in tropical Africa, occurring in a variety of forested habitats. Estimates of total surviving numbers have increased from about 200,000 in the 1980s to a maximum of almost 300,000 in 2003. However, this apparent increase comes about from new survey data, rather than representing a measured increase in actual population numbers. Infectious disease decimated several chimpanzee populations during the 1990s, and data from parts of Gabon, extrapolated to that country as a whole, suggest a major decline in great ape populations caused by disease and hunting. However, accurate data on population numbers are absent for the majority of wild chimpanzee populations. I found reports of the presence of Pan troglodytes in at least 51 national parks in at least 19 countries; some of these parks have been established very recently. Chimpanzees also occur in many non-park conservation areas. A set of large, well protected parks could safeguard chimpanzees for the foreseeable future. Although many African parks do not function well at present, mechanisms to improve their function are understood and available. By a strict application of IUCN threat criteria, P. troglodytes can be considered Endangered, based on estimated rates of past decline and on the species' long generation time. Relatively speaking, however, P. troglodytes is less endangered than are orangutans or gorillas, and the species is unlikely to go extinct by the year 2100, especially if existing conservation measures improve. The IUCN threat-rating system has become overly complex; the system can produce results that do not accord with common sense and these results must therefore be interpreted with care. PMID- 16273307 TI - Prospective study on the presentation and outcome of patients with an acute hernia. AB - Recent advances in hernia surgery should help to make operation more acceptable to patients and their doctors. The aim of this study was to prospectively assess the presentation and management of patients with an acute hernia in light of these changes. Data on all patients admitted with an acute hernia between March 2001 and February 2004 was entered on a prospective database. During the 3 year study period, of the 91 patients admitted with an acute hernia, 41 were ventral, 24 femoral, 24 inguinal and 2 parastomal. Forty-six had a previous medical assessment, 18 of these had been declared unfit for operation at that assessment; ten were ASA4 (ASA, American Society of Anaesthesiology), five ASA3 and three ASA2. Eleven patients were on the waiting list for operation, three of whom had a previous acute hospital admission. For 30 patients this hospital admission was the first indication that they had a hernia while the remaining were aware that they had a hernia but did not seek medical advice. Of the five patients who died, two while being assessed for operation and three postoperatively, three were ASA4 and had a ventral hernia while two were ASA3 with a femoral hernia. Despite advances in hernia surgery there is still room for improvement in preoperative assessment of patients presenting with an acute hernia. PMID- 16273306 TI - Cost-effective laparoscopic TEP inguinal hernia repair: the Portsmouth technique. AB - NICE (UK) has not recommended unilateral primary laparoscopic inguinal hernia repair because of its expense. A two-port technique without balloon inflator or routine tacking was developed, which helped reduce costs to just Pounds 35 more than day-case open hernia repair. Over a 6-month period, 40 patients underwent 60 TEP repairs with a 6-month follow up. Zero degree laparoscope (10 mm) and blunt graspers (5 mm) created the pre-peritoneal space, identified landmarks and completed the dissection. Trimmed 15 x 15 cm mesh was placed over each defect. Operating times for unilateral and bilateral hernias for consultants and supervised trainees were 30*, 42.5* and 40*, 55* min (*: Median) respectively. Verbal rating pain scores at 24 and 72 h were 1* (0-3) and 0* (0-2) respectively. Patients returned to activity, driving and work in 5*, 7* and 14* days respectively. Cost of laparoscopic hernia repair was calculated at Pounds 105. A two-port laparoscopic hernia repair can be performed effectively and safely, in reasonable time and at a low cost. These data support the use of this technique in primary unilateral inguinal hernia. PMID- 16273308 TI - Multiple intraosseous meningiomas. AB - In our knowledge, we are the first to report an observation on multiple osteomeningioma. PMID- 16273309 TI - Marked saggital sinus dilatation and thrombi without thrombosis in a patient with systemic lupus erythematosus. AB - Superior saggital sinus thrombosis (SSST), which has a strong causal link with antiphospholipid syndrome, rarely occurs in patients with systemic lupus erythematosus (SLE). We describe a 34-year-old woman with SLE whose clinical problem was mild headache. Her serology indicated negative antiphospholipid, anticardiolipin antibodies and lupus anticoagulants. However, marked dilatation of the entire saggital sinus with scattered thrombi was observed in enhanced-, surface- and three-dimensional reconstructed CTs (3D-CTs) without abnormal intra axial signal in brain MRI. The enhanced-, surface- and 3D-CTs are useful to detect silent dural sinus dilatation with scattered thrombi in a patient with SLE without any symptoms of SSST. PMID- 16273310 TI - Evaluating the Korean version of the Multidimensional Health Assessment Questionnaire in patients with rheumatoid arthritis. AB - Although the Health Assessment Questionnaire (HAQ) and the Modified Health Assessment Questionnaire are useful tools for assessing and monitoring patients with rheumatic diseases, they have a "floor effect" and do not fully reflect the psychological status of patients. Recently, the Multidimensional Health Assessment Questionnaire (MDHAQ) was developed to overcome these shortcomings. We translated the MDHAQ into the Korean language and evaluated its reliability and validity for use with Korean-speaking patients with rheumatoid arthritis (RA). The questionnaire was translated into the Korean language by three translators, who were aware of its objectives, and it was translated back into the English language by three different translators. One question was modified to reflect Korean culture, and imperial measures were changed to metric measures because most Koreans use the metric system. The Korean MDHAQ was administered to 136 patients with RA who were attending the outpatient rheumatology clinic at the Chonnam National University Hospital (Gwangju, South Korea). Test-retest reliability was assessed in 101 patients after 1 week. To assess criterion validity, we compared MDHAQ scores with HAQ scores and the American College of Rheumatology (ACR) functional class. To test construct validity, the MDHAQ was compared to ACR core criteria (tender and swollen joint count, pain, patient's global assessment, physician's global assessment, erythrocyte sedimentation rate, and C-reactive protein), the Beck Depression Inventory (BDI), and the State-Trait Anxiety Inventory (STAI). The test-retest reliability was analyzed by computing kappa statistics, which ranged from 0.60 to 0.76. Cronbach's alpha coefficient ranged from 0.892 to 0.938. The MDHAQ was significantly correlated with the HAQ and ACR functional class (all p<0.001). The correlations between the MDHAQ scores and the ACR core set, BDI, and STAI were all high and statistically significant. The Korean version of the MDHAQ is a reliable, valid tool for assessing Korean patients with RA. PMID- 16273311 TI - Successful closure of a bladder neck fistula complicated by urethral and vaginal stenosis, using oxidized cellulose (Surgicel) for reinforcement. AB - We describe a case of a bladder neck fistula in a 25-year-old lady presenting with true urinary incontinence, vaginal constriction and induration, with vaginal length reduced to only 1.5 cm. There was an 8-mm fistula involving upper urethra and bladder neck, with fibrosis all around. Using Schuchardt incision, the fistula was reached and mobilization tried. As there was less available tissue, a 3 x 2-cm layer of oxidized cellulose was stitched between the bladder and the vaginal mucosa for reinforcement and to achieve a watertight closure of fistula. PMID- 16273312 TI - Visualization of a large ventricular aneurysm in a young man by 16-slice multi detector row spiral computed tomography before successful surgical treatment. AB - Left ventricular (LV) aneurysm after myocardial infarction without any clinical symptoms is a rare complication of coronary artery disease. In most cases, especially in young people, this complication is diagnosed too late, after rupture into the pericardial cavity has occurred. Here we present the successful surgical repair of a large LV aneurysm in a 35-year-old man. We could visualize this aneurysm and the coronary arteries non-invasively by contrast-enhanced 16 slice multi-detector row spiral computed tomography (MDCT). PMID- 16273314 TI - Receptor-selective retinoids inhibit the growth of normal and malignant breast cells by inducing G1 cell cycle blockade. AB - Despite advances in treatment, breast cancer continues to be the second leading cause of cancer mortality in women. Statistics suggest that while focus on treatment should continue, chemopreventive approaches should also be pursued. Previous studies have demonstrated that naturally occurring retinoids such as 9 cis retinoic acid (9cRA) can prevent breast cancer in animal models. However, these studies have also shown that these compounds are too toxic for general use. Work from our laboratory showed that an RXR-selective retinoid LGD1069 prevented tumor development in animal models of cancer with reduced toxicity as compared to an RAR-selective retinoid TTNPB. In the present study, we investigated the mechanisms by which receptor-selective retinoids inhibit the growth of normal and malignant breast cells. Our results demonstrate that the synthetic retinoids tested are as effective as 9cRA in suppressing the growth of normal human mammary epithelial cells (HMECs) and estrogen receptor-positive (ER-positive) breast cancer cells. Although the receptor-selective retinoids induce minimal amounts of apoptosis in T47D breast cancer cells, the predominant factor that leads to growth arrest is G1 cell cycle blockade. Our data indicate that this blockade results from the downregulation of Cyclin D1 and Cyclin D3, which in turn causes Rb hypophosphorylation. Non-toxic retinoids that are potent inducers of cell cycle arrest may be particularly useful for the prevention of breast cancer. PMID- 16273315 TI - The application of a random effects model to censored twin data. AB - The authors propose a random effects model to analyze the latent genetic and environmental effects on determining censored outcomes in twin studies. In this model, six normally distributed random effects are used to describe the correlation within twin pairs. The authors employ a Monte Carlo Expectation Maximization approach for obtaining maximum likelihood estimates of fixed effects and the variances of random effects. The variances of the random effects are reparameterized to be equivalent to genetic and environmental effects in traditional twin models. The authors illustrate this model using data from the Vietnam Era Twin Registry to explore the magnitude of the genetic influence on twin similarity for the age of onset of alcohol abuse. Our results show genetic factors contribute about one third of twin similarity in the age of onset of alcohol abuse in male twins. The application of this model to twin data is discussed. PMID- 16273313 TI - Ototoxicity of cisplatin plus standard radiation therapy vs. accelerated radiation therapy in glioblastoma patients. AB - PURPOSE: To assess the effect of cisplatin (CDDP) plus concurrent radiation therapy on hearing loss. METHODS: 451 patients with glioblastoma multiforme (GBM) were randomly assigned after surgery to: Arm A: Carmustine (BCNU) + standard radiation therapy (SRT); Arm B: BCNU + accelerated radiation therapy (ART: 160 cGy twice daily for 15 days); Arm C: CDDP + BCNU + SRT; or Arm D: CDDP + BCNU + ART. Patients on arms C and D received audiograms at baseline, and prior to the start of RT, and prior to cycles 3 and 6. Otologic toxicities were recorded at each visit. RESULTS: 56% of patients had hearing loss at baseline. 13% and 50% of patients experienced worsening ototoxicity after 1 year of treatment in arms A and B vs. C and D, respectively, with 13% of those on arms C and D experiencing significant ototoxicity (>or= grade 3) at 6 months. Increasing age was associated with an increased risk of ototoxicity. CONCLUSIONS: Increased exposure to CDDP increases the risk of ototoxicity over time. Older patients are more susceptible to hearing loss with CDDP. The low proportion of patients with clinically significant ototoxicity suggests that baseline screening is unnecessary in GBM patients. PMID- 16273316 TI - Application of Bayesian inference using Gibbs sampling to item-response theory modeling of multi-symptom genetic data. AB - Several "genetic" item-response theory (IRT) models are fitted to the responses of 1086 adolescent female twins to the 33 multi-category item Mood and Feeling Questionnaire relating to depressive symptomatology in adolescence. A Markov chain Monte Carlo (MCMC) algorithm is used within a Bayesian framework for inference using Gibbs sampling, implemented in the program WinBUGS 1.4. The final model incorporated separate genetic and non-shared environmental traits ("A and E") and item-specific genetic effects. Simpler models gave markedly poorer fit to the observations judged by the deviance information criterion (DIC). The common genetic factor showed major loadings on melancholic items, while the environmental factor loaded most highly on items relating to self-deprecation. The MCMC approach provides a convenient and flexible alternative to Maximum Likelihood for estimating the parameters of IRT models for relatively large numbers of items in a genetic context. Additional benefits of the IRT approach are discussed including the estimation of latent trait scores, including genetic factor scores, and their sampling errors. PMID- 16273317 TI - A school-based twin study of handedness among adolescents in Taiwan. AB - This study aimed to evaluate whether twinning might influence handedness and the relative contribution of genetic and environmental factors to handedness in a total of 321 pairs of twins, 36 same-sex sib-pairs, and 1020 singletons, aged 12 16 and systematically recruited from the junior high schools in Taipei. Twins' zygosity was determined by a combination of DNA typing and physical similarity. The direction and consistency of handedness in twins did not differ from that seen in singletons. Compared with the full model containing additive genes (A), shared (C), and non-shared (E) environment, both AE and CE models had equivalently acceptable fit. The contribution from additive genes in the AE model was estimated to be 16% (directional) to 13% (consistent) for the continuous handedness and 34-10% for the categorical one, whereas the corresponding contribution from shared environment in the CE model was 14-14% and 32-11%, respectively. Handedness in adolescents appears to be not influenced by twinning and not substantially heritable, whereas environmental factors, especially those not shared between siblings, are the most important ones for explaining individual variations. PMID- 16273318 TI - Genetic covariation between event-related potential (ERP) and behavioral non-ERP measures of working-memory, processing speed, and IQ. AB - The aim of this study was to identify genetic covariants for fundamental measures of brain function (event-related potentials (ERPs): P300 latency and slow wave amplitude recorded in a working-memory task) and more complex cognitive measures (behavioral non-ERP measures: working-memory performance, information processing speed, IQ). Data were collected from 252 monozygotic and 297 dizygotic twin pairs aged 16. Multivariate modeling identified two independent genetic factors associated with processing speed that also influenced working-memory performance (one reflected the duration of neural activity required to evaluate target information, the other reflected more general cognitive and speed-related abilities). However, the allocation of neural resources, as assessed by ERP slow wave amplitude measures, was not associated with the other cognitive measures investigated. Thus, of the ERP measures examined, P300 latency, but not slow wave amplitude, may be an informative measure to include (i.e., with working-memory performance) in future multivariate linkage and association analyses of cognitive function. PMID- 16273319 TI - Opposite effects of androgen receptor CAG repeat length on increased risk of left handedness in males and females. AB - Prenatal exposure to testosterone has been hypothesised to effect lateralization by influencing cell death in the foetal brain. Testosterone binds to the X chromosome linked androgen receptor, which contains a polymorphic polyglutamine CAG repeat, the length of which is positively correlated with testosterone levels in males, and negatively correlated in females. To determine whether the length of the androgen receptor mediates the effects of testosterone on laterality, we examined the association between the number of CAG repeats in the androgen receptor gene and handedness for writing. Association was tested by adding regression terms for the length of the androgen receptor alleles to a multi factorial-threshold model of liability to left-handedness. In females we found the risk of left-handedness was greater in those with a greater number of repeats (p=0.04), this finding was replicated in a second independent sample of female twins (p=0.014). The length of the androgen receptor explained 6% of the total variance and 24% of the genetic variance in females. In males the risk of left handedness was greater in those with fewer repeats (p=0.02), with variation in receptor length explaining 10% of the total variance and 24% of the genetic variance. Thus, consistent with Witelson's theory of testosterone action, in all three samples the likelihood of left handedness increased in those individuals with variants of the androgen receptor associated with lower testosterone levels. PMID- 16273320 TI - Assortative mating in Drosophila adapted to a microsite ecological gradient. AB - Elucidating the causes of population divergence, and ultimately speciation, is a central objective of evolutionary biology. A number of previous studies of Drosophila populations from the Nahal Oren canyon (Mt Carmel, Israel) revealed significant interslope differences for a complex of fitness and behavioral traits. Peculiarities in courtship song patterns and nonrandom mating were observed, despite a small interslope distance. Single and multiple mate choice tests with D. melanogaster from the opposite slopes displayed highly significant assortative mating, with preference for sexual partners from the same slope. Here we report the results on mate choice in the sibling species D. melanogaster and D. simulans inhabiting Nahal Oren canyon. Significant assortative mating was found in both species. Genetic heterogeneity in mate choice was found among the isofemale lines of D. melanogaster. Samples of isofemale lines established from females collected in spring and fall seasons show the same mating patterns. PMID- 16273321 TI - Widespread evidence for non-additive genetic variation in Cloninger's and Eysenck's personality dimensions using a twin plus sibling design. AB - Studies using the classical twin design often conclude that most genetic variation underlying personality is additive in nature. However, studies analyzing only twins are very limited in their ability to detect non-additive genetic variation and are unable to detect sources of variation unique to twins, which can mask non-additive genetic variation. The current study assessed 9672 MZ and DZ twin individuals and 3241 of their siblings to investigate the environmental and genetic architecture underlying eight dimensions of personality: four from Eysenck's Personality Questionnaire and four from Cloninger's Temperament and Character Inventory. Broad-sense heritability estimates from best-fitting models were two to three times greater than the narrow-sense heritability estimates for Harm Avoidance, Novelty Seeking, Reward Dependence, Persistence, Extraversion, and Neuroticism. This genetic non additivity could be due to dominance, additive-by-additive epistasis, or to additive genetic effects combined with higher-order epistasis. Environmental effects unique to twins were detected for both Lie and Psychoticism but accounted for little overall variation. Our results illustrate the increased sensitivity afforded by extending the classical twin design to include siblings, and may provide clues to the evolutionary origins of genetic variation underlying personality. PMID- 16273322 TI - Genetic and environmental contributions to loneliness in adults: the Netherlands twin register study. AB - Heritability estimates based on two small studies in children indicate that the genetic contribution to individual differences in loneliness is approximately 50%. Heritability estimates of complex traits such as loneliness may change across the lifespan, however, as the frequency, duration, and range of exposure to environmental influences accrues, or as the expression of genetic factors changes. We examined data on loneliness from 8,387 young adult and adult Dutch twins who had participated in longitudinal survey studies. A measure of loneliness was developed based on factor analyses of items of the YASR (The Young Adult Self Report). Variation in loneliness was analyzed with genetic structural equation models. The estimate of genetic contributions to variation in loneliness in adults was 48%, which is similar to the heritability estimates found previously in children. There was no evidence for sex or age differences in genetic architecture. Sex differences in prevalence were significant, but we did not see an association with age or birth cohort. All resemblance between twin relatives was explained by shared genes, without any suggestion of a contribution of shared environmental factors. PMID- 16273323 TI - Left ventricular remodeling after myocardial infarction in mice with targeted deletion of the NADPH oxidase subunit gp91PHOX. AB - BACKGROUND: Oxidative stress is involved in progression of left ventricular hypertrophy and heart failure. Since NADPH oxidases are a major source of reactive oxygen species in the heart, we studied left ventricular remodeling after myocardial infarction in mice with targeted deletion of the NADPH oxidase subunit gp91(phox). METHODS AND RESULTS: gp91(phox) knockout (KO) and wild-type (WT) animals underwent coronary artery ligation. Mortality was significant higher in the gp91(phox) KO mice. However, transthoracic echocardiography performed at days 1, 7, and 56 at mid-papillary levels revealed that progression of left ventricular remodeling was not influenced by the genotype. Moreover, systemic oxidative stress was not reduced in gp91(phox) KO mice as indicated by a significant increase in lipid peroxides potentially mediated by an increase of the NADPH subunit nox-1 in gp91(phox) KO mice. CONCLUSION: Targeted deletion of the NADPH subunit gp91(phox) does not affect left ventricular remodeling following myocardial infarction and does not decrease the production of oxidative stress. However, the final role of the different NADPH subunits in the heart under pathophysiologic conditions remains to be determined. PMID- 16273324 TI - Retinoic acids increase expression of GLUT4 in dedifferentiated and hypertrophied cardiac myocytes. AB - Sufficient expression of the insulin-sensitive glucose transporter GLUT4 may be crucial for the survival of cardiac myocytes in situations of stress. Expression of GLUT4 in cardiac myocytes correlates with cell differentiation and is reduced in the hypertrophied and failing myocardium. Adult rat cardiomyocytes (ARC) in primary culture undergo dedifferentiation and reduction of GLUT4 expression. Depending on the culture condition partial redifferentiation and/or hypertrophy follows. All-trans (at) and 9-cis retinoic acids (RA) are morphogenetic agents important for cell differentiation. Both atRA and 9-cisRA restored GLUT4 expression in dedifferentiated ARC, while only 9-cisRA could increase GLUT4 expression in hypertrophic ARC. The effects of RA were associated with improved differentiation of the cardiac myocytes, as assessed from the expression of atrial natriuretic factor and the morphology of the contractile apparatus. In neonatal rat cardiomyocytes, 9-cisRA, but not atRA, stimulated transcription from the glut4 promoter. In conclusion, treatment with RA can restore the down regulated expression of GLUT4 in cardiomyocytes in association with a partial improvement of the differentiated phenotype. PMID- 16273325 TI - Is there an association between high heparanase level and osteoporosis risk in breast cancer patients without clinical evidence of bone metastases? PMID- 16273326 TI - A method for improving the management of controversial wetland. AB - Valley bottom wetlands in agricultural landscapes often are neglected in national and regional wetland inventories. Although these areas are small, located in the bottomlands of the headwater catchments, and scattered in the rural landscape, they strongly influence hydrology, water quality, and biodiversity over the whole catchment area. Valley bottom wetlands often are considered as controversial wetlands. Awareness of the functional role of wetlands is increasing, in parallel with their progressive disappearance in intensive farming landscapes. The need to improve tools for controlling wetland management is a primary consideration for decision makers and land users. This article proposes a method for the inventory of valley bottom wetlands. The method is based on the functional analysis of potential, existing, and efficient valley bottom wetlands (the PEEW approach). Several indicators are proposed for checking the validity of such an approach. Potential wetlands are delineated by means of a topographic index using topographic and pedoclimatic criteria computed from a Digital Elevation Model and easily accessible databases. Existing wetlands are identified from observed surface moisture, the presence of specific wetland vegetation, or soil feature criteria. Efficient wetlands are defined through a given function, such as flow or pollutant regulation or biodiversity control. An analysis of areas at the limits between potential, existing, and efficient wetlands highlights land cultivated or drained in the past, which currently represents negotiating areas in which rehabilitation and other intended management actions can be implemented. PMID- 16273327 TI - Cost-effective control of interdependent water pollutants. AB - Excess nutrient loads to coastal waters lead to increased production of algae, which when decaying cause oxygen depletion in bottom sediments, which in turn leads to major changes in marine ecosystems. A cost-minimization model was investigated in which nitrogen and phosphorus were assumed to interact with respect to algae production. It was investigated under what conditions only a single nutrient should be decreased and when it is cost-effective to decrease both nutrients. The model was applied to the Baltic Proper, which is the largest of the seven major basins in the Baltic Sea. Results showed that the stringency of the environmental target, as well as assumptions regarding substitutability between nutrients, are important factors to determine the nutrient on which to focus. Uniform decrease rates for emissions are often used in international agreements, and the results of this model showed that the costs of making such proportional decreases could be substantial. PMID- 16273329 TI - Pelvic organ prolapse and urinary incontinence in women with surgically managed rectal prolapse: a population-based case-control study. AB - PURPOSE: This study aimed to investigate the prevalence of genital prolapse surgery and urinary incontinence in female patients operated on for rectal prolapse compared with a matched control group without rectal prolapse. METHODS: Fifty-two patients with a history of abdominal rectal prolapse surgery and 200 randomly selected age-matched and gender-matched control subjects without rectal prolapse received an extensive health care history survey. RESULTS: Response rate in the patient group was 48 of 52 (92 percent) and 165 of 200 (82 percent) in the control group. Rectal prolapse was associated with an increased risk of surgery for uterine prolapse (odds ratio = 3.1; 95 percent confidence interval = 1.4-6.9) and vaginal wall prolapse (odds ratio = 3.2; 95 percent confidence interval = 1.3 7.8). Mean age at hysterectomy because of uterine prolapse was 54.7 years in the patient group compared with 62.6 years in the control group (P < 0.01). Mean age at vaginal wall prolapse surgery was 60.2 years in the patient group compared with 66.6 years in the control group (P < 0.05). There were no significant differences between the cohorts regarding prevalence or age at debut of urinary incontinence. CONCLUSION: Our results indicate a strong association between rectal and genital prolapse surgery suggesting that diagnosis of rectal prolapse necessitating surgical intervention should prompt a multidisciplinary pelvic floor assessment. PMID- 16273330 TI - Morphologic alterations associated with mechanical bowel preparation before elective colorectal surgery: a randomized trial. AB - PURPOSE: The feasibility and safety of left-sided colorectal procedures with avoidance of mechanical bowel preparation has recently been demonstrated. Moreover, mechanical preparation has been associated with an increased risk for abdominal septic complications, including anastomotic leakage. This study was designed to determine whether mechanical bowel preparation is associated with histologic alterations in the colon. METHODS: Fifty patients (mean age, 61 (range, 45-78) years) scheduled to undergo elective colorectal surgery were prospectively randomized to receive mechanical preparation (polyethylene glycol; Group 1) or no preparation (Group 2) preoperatively. A macroscopically healthy segment of the bowel was excised at the proximal margin of the colectomy piece. A pathologist, blinded to the patient's group allocation, assessed various morphologic parameters. RESULTS: Indications for colectomy (cancer and complicated diverticulosis) did not differ between groups. Bowel wall alterations were more frequent in patients who received a preparation. The most striking alterations associated with mechanical preparation were loss of superficial mucus (moderate-to-severe in 96 and 52 percent in Groups 1 and 2, respectively; P < 0.001) and epithelial cells (moderate-to-severe in 88 and 40 percent in Groups 1 and 2, respectively; P < 0.01). In addition, inflammatory changes, i.e., lymphocytes (severe in 48 and 12 percent in Groups 1 and 2, respectively; P < 0.02) and polymorphonuclear cells infiltration (severe in 52 and 8 percent in Groups 1 and 2, respectively; P < 0.02), were more prevalent after mechanical preparation. CONCLUSIONS: Mechanical bowel preparation is associated with structural alteration and inflammatory changes in the large bowel wall. Although bowel wall inflammation is a known risk factor for anastomotic leak, it remains to be elucidated whether these changes have a direct relation to the deleterious effect of mechanical bowel preparation in terms of abdominal morbidity. PMID- 16273331 TI - Minimal access colorectal surgery: is it truly minimally invasive? PMID- 16273332 TI - Granulocyte colony-stimulating factor therapy and systemic inflammation: effect on interleukin-10. PMID- 16273333 TI - Effects of cyclooxygenase inhibition on bone, tendon, and ligament healing. AB - Cyclooxygenases (COX-1 and COX-2) catalyze the conversion of arachidonic acid to prostaglandins (PGs). PGs play a significant role in bone metabolism in health and disease. Conventional non-selective, non-steroidal anti-inflammatory drugs (NSAIDs) and selective COX-2 inhibitors (COXIBs) are widely used in patients with musculoskeletal conditions and as a post-surgical analgesics. Due to their effects on PG synthesis, these drugs have been hypothesized to affect the healing process of bone fractures and orthopedic surgical sites. A variety of experimental models of bone, ligament, and tendon repair have assessed the effects of these selective and non-selective COX inhibitors in animals, but with variable outcomes. At this time, large-scale, robust clinical study data do not exist, limiting the relevant assessment of experimental animal data to humans. Here, we provide a critical review of available data on the role of PGs and the effect of COX inhibitors on bone, tendon, and ligament repair. Collectively, this assessment suggests potential involvement of PGs in the healing process of these tissues via modulation by non-selective NSAIDs and selective COX-2 inhibitors. PMID- 16273334 TI - Histamine H1 receptor occupancy and pharmacodynamics of second generation H1 antihistamines. AB - The predictive efficacy of drugs in humans is frequently estimated from both a high affinity for their receptor as measured in vitro and a long plasmatic half life. This is grossly misleading since one key parameter is missing: drug concentration at the receptor site in vivo. As a case study we compared the efficacies of three H(1) antihistamines in inhibiting histamine-induced wheal and flare in humans at two different time points with the above mentioned parameters. It is concluded that estimating in vivo receptor occupancy, which takes into account both the affinity of the drug for the receptor and its free plasma concentration, is a far better predictor for human pharmacodynamics and hence antihistamine potency, than considering in vitro affinity and plasmatic half-life only. PMID- 16273335 TI - Mast cell hyperplasia induced by multiple challenge with cedar pollen in sensitised guinea pigs. AB - OBJECTIVE AND DESIGN: Because immediate allergic symptoms are aggravated by repetitive antigen challenges in a guinea pig model of Japanese cedar pollen induced conjunctivitis, we determined whether conjunctival mast cells are different in number between the acute and chronic stages. METHODS: Sensitised guinea pigs were challenged by dropping a pollen suspension into their eyes once a week. Conjunctival tissue sections were stained with toluidine blue. Ophthalmic lavage was performed to assay for mast cell mediators. RESULTS: At the 20th and 40th challenges, the number of mast cells increased by 4- to 5-fold compared with the 1st challenge. Although mast cell degranulation was insignificant 10 min after the 1st challenge, the 20th and 40th challenges produced significant degranulation. After multiple challenges, the amount of histamine and tryptase like activity in the lavage fluid was dramatically increased. CONCLUSION: Increased mast cells are associated with aggravated symptoms. Mast cell mediators may be involved in pathogenesis at the chronic stage. PMID- 16273336 TI - Release of TNF-alpha and IL-1beta from porcine brain endothelium corresponds to the pyrogenic potential of three marketed formulations of amphotericin. AB - BACKGROUND: Formulations of amphotericin include a deoxycholate suspension (d Amph), an amphotericin-B lipid complex (Ablc), and a liposomal product (L-Amph). Fever is most frequent with d-Amph, intermediate with Ablc, and lowest with L Amph. OBJECTIVE: To determine if the release of tumor necrosis factor-alpha (TNF alpha) and interleukin-1-beta (IL-1) from brain endothelium corresponds to the incidence of amphotericin fever. RESULTS: Release of TNF-alpha and IL-1beta after L-Amph treatment was similar to negative controls while after d-Amph treatment release was similar to lipopolysaccharide. Ablc treatment produced intermediate pyrogen release.NF-kappaB expression, a transcriptional regulator for TNF-alpha and IL-1beta genes, corresponded to this secretion pattern. TNF-alpha release was elevated 2 hours (p = 0.0021) after treatment while significant elevations in IL 1beta required 6 hours (p = 0.0009). CONCLUSION: Results from this in vitro study suggest that amphotericin fever may be directly mediated by brain endothelium. These experiments also suggest that amphotericin fever is initially mediated by TNF-alpha. PMID- 16273337 TI - Human immunodeficiency virus-1 Tat protein up-regulates interleukin-6 and interleukin-8 expression in human breast cancer cells. AB - OBJECTIVE: To examine whether HIV-1 Tat protein increases the metastatic potential of human breast cancer cells through induction of pro-inflammatory tumor microenvironment. METHODS: Real-time RT-PCR and ELISA were employed to determine the mRNA and protein expression of IL-6 and IL-8 in highly metastatic human breast cancer cell line, MDA-MB-231. To investigate the transcriptional regulatory mechanisms of Tat-mediated up-regulation of IL-6 and IL-8, EMSA and reporter gene assay were carried out. RESULTS: Exposure of MDA-MB-231 cells to Tat resulted in a significant and dose-dependent up-regulation of IL-6 and IL-8 mRNA and protein expression. HIV-1 Tat protein also markedly increased NF-kappaB DNA-binding activity and transactivation in MDA-MB-231 cells. Additionally, pretreatment with NF-kappaB inhibitors significantly attenuated the ability of Tat to up-regulate IL-6 and IL-8 expression. It was also found that exposure of MDA-MB-231 cells to Tat induced up-regulation of MMP-9 expression at both mRNA and protein levels. CONCLUSIONS: These results suggest that HIV-1 Tat protein up regulates expression of IL-6 and IL-8 in human breast cancer cells by NF-kappaB dependent pathway. These data may contribute to exploration of the new molecular mechanisms leading to novel approaches for the therapeutic drug developments specifically targeted against the inflammatory pathways of breast cancer metastasis in AIDS patients. PMID- 16273338 TI - Riboflavin in innate and acquired immune responses. AB - OBJECTIVE AND DESIGN: Riboflavin, also known as vitamin B2, is a micronutrient with a key role in maintaining human health. It has also been shown to enhance host resistance to bacterial infections in mice. The aim of this study was to assess the role of vitamin B2 treatment in inflammatory conditions. SUBJECTS AND METHODS: Three models of inflammatory states were assessed. One of them encompasses neutrophil mediated but T cell/macrophage independent cutaneous inflammation. Another one is delayed type hypersensitivity reaction (DTH), a T cell/macrophage dependent but neutrophil independent inflammatory response. The third one is collagen- induced arthritis, having components from both of the above described reactions. Mice were treated with vitamin B2, administered by peritoneal injections, throughout the course of the experiments. RESULTS: The granulocyte dependent reaction to olive oil was significantly reduced in vitamin B2 treated mice. In contrast, DTH reactivity and collagen II arthritis were not affected by the treatment. CONCLUSION: Riboflavin administration affects neutrophil migration but does not alter acquired immune responsiveness. PMID- 16273339 TI - High-performance liquid chromatography-tandem mass spectrometry method for quantifying sulfonylurea herbicides in human urine: reconsidering the validation process. AB - We have developed a method for measuring 17 sulfonylurea (SU) herbicides in human urine. Urine samples were extracted using solid phase extraction (SPE), pre concentrated, and analyzed by high-performance liquid chromatography-tandem mass spectrometry using turboionspray atmospheric pressure ionization. Carbon 13 labeled ethametsulfuron methyl was used as an internal standard. Chromatographic retention times were under 7 minutes. Total throughput was estimated as >100 samples per day. Because only one labeled internal standard was available for the analysis, we were forced to reconsider and restructure the validation process to include stringent stability tests and analyses of urine matrices of differing compositions. We describe our restructured validation process and the critical evaluation it provides for the method developed. The limits of detection (LOD) ranged from 0.05 microg/L to 0.10 microg/L with an average LOD of 0.06 microg/L. Average total relative standard deviations were 17%, 12% and 8% at 0.1 microg/L, 3.0 microg/L and 10 microg/L, respectively. Average extraction efficiencies of the SPE cartridges were 87% and 86% at 2.5 microg/L and 25 microg/L, respectively. Chemical degradation in acetonitrile and urine was monitored over 250 days. Estimated days for 10% and 50% degradation in urine and acetonitrile ranged from 0.7 days to >318 days. The influence of matrix effects on precision and accuracy was also explored. PMID- 16273341 TI - [Therapeutically induced arteriogenesis in the brain. A new approach for the prevention of cerebral ischemia with vascular stenosis]. AB - Stroke is the leading cause of disability and a major cause of death in Germany and the western world. Ischemic stroke involves different pathophysiologic mechanisms such as thromboembolic vascular occlusion, cerebral micro- or macroangiopathy, extracranial arterial stenosis, and cardiac embolism. Experimental and clinical studies have shown that arteriogenesis, the adaptive growth of pre-existing collateral arteries, can be therapeutically enhanced in peripheral circulation and the heart. We examined the consequences to time course and hemodynamics of brain arteriogenesis in a chronic hypoperfusion model following systemic administration of the hemopoietic growth factor called granulocyte macrophage colony stimulating factor (GM-CSF). Treatment with GM-CSF led to the growth of intracranial collateral arteries, which improved the cerebral hemodynamic reserve and significantly reduced energy failure when brains were additionally challenged by hypotension. Therapeutically induced arteriogenesis may be of considerable interest for preventing infarction in patients with uncompensated cerebrovascular disease. PMID- 16273340 TI - [Present status and future possibilities of adjuvant pharmacotherapy for aphasia]. AB - Aphasia is one of the most frequent and disabling consequences of stroke. Poor spontaneous recovery and the limited success of conventional speech therapy bring up the question of how current treatment approaches can be improved. Besides increasing training frequency-with daily sessions lasting several hours and high repetition rates of language materials ("massed training")-adjuvant drug therapy may help to increase therapy efficacy. In this article, we illuminate the potential of monoaminergic (bromocriptine, levodopa, d-amphetamine) and cholinergic (donepezil) substances for treating aphasia. For a final evaluation of combined massed training and adjuvant pharmacotherapy, randomized, placebo controlled (multicenter) clinical trials with sufficient numbers of patients are needed. Furthermore, results of experimental animal studies of functional recovery in brain damage raise hopes that neurotrophic factors or stem cells might find a place in recovery from aphasia in the intermediate future. PMID- 16273342 TI - Does the English NHS have a 'Health Benefit Basket'? AB - A 'health benefit basket' is a range of publicly entitled health-related goods and services. Primary legislation ensures the provision of broad categories of health care, but this provision is subject to political discretion. Case law has established that health care organisations may not operate a 'blanket ban' for particular services. This means that the English health basket currently has very few specific services explicitly included or excluded. Regulation may, however, be important in determining citizens' rights. With reference to 'services of curative care', this paper explores whether the NHS is moving towards a more explicit definition of a health basket. PMID- 16273343 TI - Coffee and tomato share common gene repertoires as revealed by deep sequencing of seed and cherry transcripts. AB - An EST database has been generated for coffee based on sequences from approximately 47,000 cDNA clones derived from five different stages/tissues, with a special focus on developing seeds. When computationally assembled, these sequences correspond to 13,175 unigenes, which were analyzed with respect to functional annotation, expression profile and evolution. Compared with Arabidopsis, the coffee unigenes encode a higher proportion of proteins related to protein modification/turnover and metabolism-an observation that may explain the high diversity of metabolites found in coffee and related species. Several gene families were found to be either expanded or unique to coffee when compared with Arabidopsis. A high proportion of these families encode proteins assigned to functions related to disease resistance. Such families may have expanded and evolved rapidly under the intense pathogen pressure experienced by a tropical, perennial species like coffee. Finally, the coffee gene repertoire was compared with that of Arabidopsis and Solanaceous species (e.g. tomato). Unlike Arabidopsis, tomato has a nearly perfect gene-for-gene match with coffee. These results are consistent with the facts that coffee and tomato have a similar genome size, chromosome karyotype (tomato, n=12; coffee n=11) and chromosome architecture. Moreover, both belong to the Asterid I clade of dicot plant families. Thus, the biology of coffee (family Rubiacaeae) and tomato (family Solanaceae) may be united into one common network of shared discoveries, resources and information. PMID- 16273344 TI - The IA-2 interactome. AB - AIMS/HYPOTHESIS: Islet antigen-2 (IA-2), a major autoantigen in type 1 diabetes, is an enzymatically inactive member of the transmembrane protein tyrosine phosphatase (PTP) family. IA-2 is located in dense-core secretory vesicles and is involved in the regulation of insulin secretion. The present experiments were initiated to identify those proteins that interact with IA-2 (i.e. the IA-2 interactome) as a first step towards elucidating the mechanism(s) by which IA-2 influences insulin secretion and serves as an autoantigen. MATERIALS AND METHODS: To determine the proteins with which IA-2 interacts, a yeast two-hybrid system was used to screen a human foetal library, and deletion mutants were used to determine the binding sites. Positive interactions were confirmed by immunoprecipitation pull-down experiments using cell lysate from transfected mammalian cell lines. RESULTS: Six new interacting proteins were identified by this approach: mitogen-activated protein kinase-activating death domain (MADD), the MADD isoform IG20, PTPrho, PTPsigma, sorting nexin 19 (SNX19) and cyclophilin A. Using a series of IA-2 deletion mutants, we identified the regions on the IA-2 molecule to which five of the interacting proteins bound. Amino acids 744-979 of IA-2 were required for the maximum binding of MADD, IG20 and SNX19, whereas amino acids 602-907 of IA-2 were required for the maximum binding of PTPrho and PTPsigma. Pull-down experiments with cell lysate from transfected mammalian cells confirmed the binding of the interacting proteins to IA-2. CONCLUSIONS/INTERPRETATION: The IA-2 interactome based on, pull-down experiments, currently consists of 12 proteins. The identification of these interacting proteins provides clues as to how IA-2 exerts its biological functions. PMID- 16273346 TI - Mealtime inhaled insulin lowers fasting glucose: a look at possible explanations. PMID- 16273345 TI - Dynamic strength training improves insulin sensitivity and functional balance between adrenergic alpha 2A and beta pathways in subcutaneous adipose tissue of obese subjects. AB - AIMS/HYPOTHESIS: The aim of this study was to investigate whether dynamic strength training modifies the control of lipolysis, with particular attention paid to the involvement of the antilipolytic adrenergic alpha 2A receptor (ADRA2A) pathway. METHODS: Twelve obese men (age: 47.4+/-2.8 years; BMI: 32.7+/ 0.9) were investigated during a 210-min euglycaemic-hyperinsulinaemic clamp conducted before and after 3 months of dynamic strength training. Before and during the third hour of the clamp, the lipolytic effect of a perfusion of isoproterenol or adrenaline (epinephrine) alone or associated with the ADRA2A antagonist phentolamine was evaluated using the microdialysis method of measuring extracellular glycerol concentration (EGC) in subcutaneous abdominal adipose tissue (SCAAT). In addition, biopsies of SCAAT were carried out before and after training to determine mRNA levels RESULTS: The training increased insulin sensitivity in adipose tissue. The decrease of EGC was more pronounced during the clamp conducted after the training period than during the clamp done in pre training conditions. Before and after the training, catecholamines induced an increase in EGC, the increase being lower during the clamp on each occasion. The isoproterenol-induced increase in EGC was higher after the training. Adrenaline induced lipolysis was potentiated by phentolamine after but not before the training. There were no training-induced changes in mRNA levels of key genes of the lipolytic pathway in SCAAT. CONCLUSIONS/INTERPRETATION: In obese subjects, dynamic strength training improves whole-body and adipose tissue insulin responsiveness. It increases responsiveness to the adrenergic beta receptor stimulation of lipolysis and to the antilipolytic action of catecholamines mediated by ADRA2As. PMID- 16273347 TI - Comment on: McClung JA, Naseer N, Saleem M et al (2005) circulating endothelial cells are elevated in patients with type 2 diabetes mellitus independently of HbA1c. Diabetologia 48:345-350. PMID- 16273348 TI - Effects of the nitric oxide donor, sodium nitroprusside, on resting leg glucose uptake in patients with type 2 diabetes. AB - AIMS/HYPOTHESIS: Nitric oxide (NO) has been implicated as an important signalling molecule in the contraction-mediated glucose uptake pathway and may represent a novel strategy for blood glucose control. The current study sought to determine whether acute infusion of the NO donor, sodium nitroprusside (SNP), increases leg glucose uptake at rest in patients with type 2 diabetes. METHODS: Fifteen male patients with type 2 diabetes (aged 54+/-4 years, mean+/-SD) were entered into a randomised, cross-over design study, examining the effect of a 30-min intra femoral infusion of SNP on leg glucose uptake. Comparison was made with a 30-min infusion of verapamil, titrated to elicit similar leg blood flow responses to SNP. Leg blood flow was measured by thermodilution in the femoral vein, and leg glucose uptake was calculated as the product of leg blood flow and the femoral arterio-venous (A-V) glucose concentration gradient. RESULTS: The two drugs increased leg blood flow to a similar extent (p=0.50). Both leg A-V glucose concentration gradient (SNP 0.12+/-0.05, verapamil -0.06+/-0.04 mmol/l; mean+/- SEM, p=0.03) and leg glucose uptake (SNP 0.17+/-0.09, verapamil -0.09+/-0.06 mmol/min; p=0.03) were higher with the SNP treatment than with verapamil. These results occurred independently of any significant difference in plasma insulin concentration between drugs (p=0.56). CONCLUSIONS/INTERPRETATION: Acute infusion of SNP resulted in greater glucose uptake relative to verapamil. NO may therefore be an important mediator of peripheral glucose disposal and a potential therapeutic target in patients with type 2 diabetes. PMID- 16273350 TI - NK cell activation by dendritic cell vaccine: a mechanism of action for clinical activity. AB - Recent reports revealed that dendritic cell (DC)-natural killer (NK) cell interaction plays an important role in tumor immunity, but few DC vaccine studies have attempted to evaluate the non-specific, yet potentially clinically relevant, NK response to immunization. In this study, we first analyzed in vitro activation of NK cells by DCs similar to those used in clinical trials. Subsequently, NK cell responses were analyzed in a phase I clinical trial of a vaccine consisting of autologous DCs loaded with a fowlpox vector encoding CEA. The data were compared with the clinical outcome of the patients. DC enhances NK activity in vitro, partly by sustaining NK cell survival and by enhancing the expression of NK-activating receptors, including NKp46 and NKG2D. Among nine patients in our clinical trial, NK cytolytic activity increased in four (range 2.5-5 times greater lytic activity) including three who had increased NK cell frequency, was stable in two and decreased in three. NKp46 and NKG2D expression showed a good correlation with the patients' NK activity. When patients were grouped by clinical activity (stable disease/no evidence of disease (stable/NE, n=5) vs progressive disease (N=4) at 3 months), the majority in the stable/NE group had increases in NK activity (P=0.016). Anti-CEA T cell response was enhanced in all the nine patients analyzed, but was not significantly different between the two groups (P=0.14). Thus, NK responses following DC vaccination may correlate more closely with clinical outcome than do T cell responses. Monitoring of NK response during vaccine studies should be routinely performed. PMID- 16273349 TI - Increased expression and activity of heme oxygenase-2 in pregnant rat aorta is not involved in attenuated vasopressin-induced contraction. AB - Pregnancy is associated with attenuated vascular reactivity to a variety of contractile agonists. Heme oxygenases are expressed in the placenta, and it has been suggested that the heme oxygenase/carbon monoxide (HO/CO) pathway plays a significant role in regulating blood flow through the feto-placental unit. In this study we investigated the possible involvement of heme oxygenases in the reduced vascular reactivity associated with pregnancy. Arginine vasopressin (AVP) (10(-10)-3x10(-7) M) induced concentration-dependent contraction of aortic ring segments from non-pregnant and pregnant (16-19 days) rats. Pregnancy did not alter the sensitivity to AVP (pD2=8.5+/-0.1 and pD2=8.4+/-0.2 in non-pregnant and pregnant rats, respectively) but significantly reduced the maximum response (107.9+/-12.7% and 38.6+/-7.4%, respectively, relative to noradrenaline-induced contraction). Western blot analysis revealed the expression of HO-2 but not HO-1 isoform in both groups. There was a significant increase in the expression and activity of HO-2 protein in aortic tissues from pregnant rats compared with those from age-matched non-pregnant rats. In the presence of L-NAME to inhibit nitric oxide (NO) synthesis, tin protoporphyrin IX (SnPP-IX, 10(-5) M), an inhibitor of heme oxygenase, did not significantly affect AVP-induced contraction in aorta segments from pregnant and non-pregnant rats. It was concluded that, though pregnancy increased the expression and activity of HO-2 in the aorta, HO-2 was not involved in the attenuated response to AVP. PMID- 16273351 TI - Monoclonal antibody A7 coupled to magnetic particles as a contrast enhancing agent for magnetic resonance imaging of human colorectal carcinoma. AB - BACKGROUND: Local recurrence, the most frequent pattern of recurrence of rectal carcinoma, is almost always fatal. The difficulty of diagnosing local recurrence contributes importantly to the poor prognosis. METHODS: We coupled monoclonal antibody (Mab) A7, which reacts specifically with human colorectal carcinoma, to ferromagnetic lignosite (FML) particles to distinguish rectal carcinoma from other tissues by magnetic resonance (MR) imaging. We examined retention of immunoreactivity by the A7-FML complexes in vitro, and also their distribution in vivo according to radiolabeling and MR imaging when injected into nude mice bearing human colorectal carcinoma xenografts. RESULTS: A7-FML retained binding activity nearly identical to that of Mab A7. Significantly more (125)I-labeled A7 FML accumulated in engrafted tumors than did (125)I-labeled normal mouse IgG-FML complexes (P < 0.05). A7-FML disappeared rapidly from the blood. Normal tissues accumulated less (125)I-labeled A7-FML than tumors; this accumulation decreased linearly with time. In MR imaging, signal intensity was reduced in the tumor by the injection of A7-FML. CONCLUSIONS: A7-FML is potentially useful as a MR contrast enhancing agent for human colorectal carcinoma xenografts implanted subcutaneously. PMID- 16273352 TI - An intracellular antibody can suppress tumorigenicity in hepatitis B virus X expressing cells. AB - Although the hepatitis B virus X protein (HBx) is thought to play a causative role in the development of hepatocellular carcinoma, it is not yet known whether interfering with HBx function may affect the cellular transformation of HBx expressing tumor cells. To address this question, we adopted an intracellular antibody fragment expression approach to block the function of HBx. Expression of a single-chain variable fragment (scFv) specific to HBx (designated as H7scFv) inhibited HBx-dependent cellular transactivation. Furthermore, H7scFv suppressed the growth of HBx-expressing tumor cells in both soft agar and nude mice. The suppressive effect of H7scFv on tumorigenicity appeared not to be mediated by inhibition of HBx-induced growth stimulation since the growth rate of these cells was not affected significantly by H7scFv expression. In conclusion, these data suggest that the HBx-dependent transformed phenotype is reversible and that HBx may be a good molecular target for the treatment of HBV-related tumors. PMID- 16273353 TI - Resistance to endocrine therapy in breast cancer. AB - Endocrine therapy is the treatment of choice for patients with breast cancer expressing estrogen receptor (ER) and/or progesterone receptor. The efficacy of endocrine therapy is well established in the prevention, adjuvant and metastatic settings. However, either de novo or acquired resistance is frequently observed. Much effort has been made to elucidate the mechanisms of action underlying resistance to endocrine therapy in breast cancer, and several possible explanations have been suggested. Our previous studies have indicated that combined treatment with an antiestrogen, fulvestrant, and an inhibitor of the HER2 signaling pathway, trastuzumab, or an inhibitor of the HER1 signaling pathway, gefitinib, leads to an additive antitumor effect in breast cancer cells expressing ER and HER2 or HER1, respectively. It has also been suggested that the HER1 or HER2 signaling pathway is upregulated during the development of antiestrogen-resistant growth in breast cancer cells. These findings suggest that signal transduction inhibitors are effective for the treatment of antiestrogen resistant breast cancer. A hypoxic microenvironment has been shown to promote malignant progression in cancer cells. Our previous study and others have suggested that hypoxia posttranscriptionally reduces ER expression and decreases sensitivity to hormonal agents in breast cancer cells. Our preliminary study has also shown that a hypoxic cytotoxin, tirapazamine, increases ER expression in breast cancer xenografts. Differential antitumor activity of tirapazamine on tumor cells under normoxic or hypoxic conditions may cause this phenomenon. These findings suggest that hypoxic cytotoxins may retard the development of endocrine resistance induced by hypoxia. Molecular mechanisms responsible for endocrine resistance in breast cancer are reviewed and possible therapeutic strategies against this resistance are discussed. PMID- 16273354 TI - Beyond simple castration: targeting the molecular basis of treatment resistance in advanced prostate cancer. AB - Over the past 20 years, research on hormonal treatments for prostate cancer focused on maximizing androgen ablation through combination therapy. Unfortunately, maximal androgen ablation increases treatment-related side effects and expense and has not significantly prolonged time to androgen-independent (AI) progression. Intermittent androgen suppression (IAS) is based on the hypothesis that if tumor cells surviving androgen withdrawal can be forced along a normal pathway of differentiation by androgen replacement, then apoptotic potential might be restored, androgen dependence may be prolonged and progression to androgen independence may be delayed. Observations from animal model studies suggest that progression to androgen independence is delayed by IAS and this strategy is now being evaluated in phase III trials. Another strategy for improving therapies in advanced prostate cancer involves targeting genes that are activated by either androgen withdrawal or chemotherapy to delay or prevent the emergence of the resistant AI phenotype. Targeted inhibition of stress-associated increases in gene expression precipitated by androgen withdrawal or chemotherapy may enhance treatment-induced apoptosis and delay progression to AI disease. Proteins fulfilling these criteria include antiapoptotic members of the Bcl-2 protein family, clusterin, Hsp27, and IGFBP-2 and IGFBP-5. The purpose of this paper is to review the rationale and progress in using targeted gene therapies to enhance tumor cell death after androgen withdrawal or taxane chemotherapy. Antisense oligonucleotides offer one approach to target genes involved in cancer progression, especially those not amenable to small molecule or antibody inhibition. The current status and future direction of several antisense oligonucleotides that have potential clinical use in cancer are also reviewed. PMID- 16273355 TI - A self-renewal assay for cancer stem cells. AB - Cancers of epithelial origin are responsible for the majority of cancer-related deaths in the USA. Unfortunately, although chemotherapy and/or radiation therapy can sometimes shrink tumors, metastatic cancers of epithelial origin are essentially incurable. It is clear that new approaches are needed to treat these diseases. Although cancer cell lines provide invaluable information, their biological properties often differ in crucial ways from de novo cancer cells. Our laboratory has developed a novel mouse model that reliably permits individual cancer cells isolated directly from patients' tumors to be assayed. This will allow the characterization of crucial signaling pathways involved in processes such as self-renewal that are critical for tumor formation by the cancer cells within de novo tumors. These tools should lead to new insights into the cellular and molecular mechanisms that drive human breast cancer growth and invasion. PMID- 16273356 TI - Estrogen signaling and prediction of endocrine therapy. AB - Estrogen plays an important role in the growth and progression of human breast cancer. Understanding the whole picture of estrogen signaling is a very important goal towards clarifying the biology of this disease. On the other hand, hormonal therapy for breast cancer has been progressing rapidly with the advent of drugs such as selective estrogen receptor (ER) modulators and aromatase inhibitors. Prediction of individual response to these hormonal therapies is becoming important for the management of breast cancer patients. To help address these basic and clinical issues, we are developing several new tools such as the focused microarray and the green fluorescent protein-reporter cell system. We first carried out expression profiling of approximately 10,000 genes in ER positive breast cancer cells. Based on the results, estrogen-responsive genes (ERG) were selected and a custom-made cDNA microarray consisting of 200 genes from a narrowed-down subset was produced. Using this microarray, we investigated various aspects of estrogen signaling such as the effect of estrogen-antagonists on ERG expression profile and functional analysis of ERbeta and novel estrogen responsive gene EGR3. Furthermore, expression levels of several candidate genes selected from the custom-array contents were analyzed by real-time RT-PCR and immunohistochemistry using breast cancer tissues to determine novel predictive factors for responsiveness to hormone therapy in primary breast cancer patients. Expression of several genes, such as HDAC6, significantly correlated with disease free and overall survival of patients treated with adjuvant tamoxifen therapy. We are currently developing a new tool for analyzing the effects of novel aromatase inhibitors in individual breast cancer patients using estrogen-responsive element green fluorescent protein-indicator cells. We hope that these approaches may provide not only new clues for elucidation of estrogen-dependent growth mechanisms of cancer, but also clinical benefits to patients by assessment of individual responses to endocrine therapy. PMID- 16273357 TI - Trastuzumab: updates and future issues. AB - Trastuzumab has had an enormous impact on the clinical management of breast cancer: the survival of Her-2-positive metastatic breast cancer patients has improved significantly and tumor Her-2 status has been built into the decision making tree for primary breast cancer patients. Several pioneering studies have shown that trastuzumab-combined chemotherapy elicits high levels of pathological complete response in the neoadjuvant setting. Currently, therefore, a more precise understanding of the mechanisms of therapeutic response is needed so that trastuzumab-based therapies can be optimized more individually. It might also be important to investigate, with greater depth, the interaction between the Her axis and the hormone-axis. This short review describes and discusses these topics. PMID- 16273358 TI - Current status of dose-dense chemotherapy for breast cancer. AB - In an effort to improve the effectiveness of chemotherapy for breast cancer, examination of the impact of dose intensity, dose density, and treatment duration may have as much relevance as the specific antineoplastic agents utilized. After several years of pilot feasibility studies of dose-dense chemotherapy regimens, whose delivery has been made safe and feasible by the use of hematopoietic growth factor support (in particular, filgrastim), we now have phase III data demonstrating the advantages of this approach in the adjuvant treatment of breast cancer. PMID- 16273359 TI - Advanced concepts in estrogen receptor biology and breast cancer endocrine resistance: implicated role of growth factor signaling and estrogen receptor coregulators. AB - Estrogen receptor (ER), mediating estrogen-signaling stimuli, is a dominant regulator and a key therapeutic target in breast cancer etiology and progression. Endocrine therapy, blocking the ER pathway, is one of the most important systemic therapies in breast cancer management, but de novo and acquired resistance is still a major clinical problem. New research highlights the role of both genomic and nongenomic ER activities and their intimate molecular crosstalk with growth factor receptor and other signaling kinase pathways in endocrine resistance. These signaling pathways, when overexpressed and/or hyperactivated, can modulate both activities of ER, resulting in endocrine resistance. Thus, these signal transduction receptors and signaling molecules may serve as both predictive markers and novel therapeutic targets to circumvent endocrine resistance. Compelling experimental and clinical evidence suggest that the epidermal growth factor/HER2/neu receptor (EGFR/HER2) pathway might play a distinct role in endocrine resistance, and especially in resistance to selective estrogen receptor modulators (SERMs) such as tamoxifen. Results from preclinical studies of treatment combinations with various endocrine therapy drugs together with several potent anti-EGFR/HER2 inhibitors are very promising, and clinical trials to see whether this new strategy is effective in patients are now ongoing. PMID- 16273360 TI - Clinical significance of estrogen receptor beta in breast cancer. AB - Ever since the estrogen receptor (ER) beta was discovered in 1996, we have been trying to determine its value as a prognostic and/or predictive factor in breast cancer and its potential as a novel target for pharmacological intervention. Recent progress in cellular experiments has shown that ERbeta works as counter partner of ERalpha through inhibition of the transactivating function of ERalpha by heterodimerization, distinct regulation on several specific promoters by ERalpha or ERbeta, and ERbeta-specific regulated genes which are probably related to its anti-proliferative properties. Accumulated data from protein studies in breast cancer tissues indicate that positive expression of ERbeta appears to correlate with a favorable prognosis. Although the number of studies is small, a positive response to tamoxifen treatment is observed in both ERalpha- and ERbeta positive populations. The significance of ERbeta2/cx, a splicing variant of ERbeta, remains controversial and needs to be analyzed in further studies. We postulate that a combined evaluation of ERbetacx with progesterone receptor may help the stratification of ERalpha-positive breast cancer. Epidemiological studies of hormone replacement therapy and isoflavone (genistein) consumption indicate the possible contribution of ERbeta-specific signaling in breast cancer prevention. A selective estrogen receptor modulator, which works as an antagonist of ERalpha and an agonist of ERbeta, may be a promising chemo-preventive treatment. PMID- 16273362 TI - New concepts of treatment strategies for hormone-related cancer. Proceedings of the 20th Bristol-Myers Squibb Nagoya International Cancer Treatment Symposium. March 11-12, 2005. Nagoya, Japan. PMID- 16273361 TI - Drug resistance in chemotherapy for breast cancer. AB - Recent developments with chemotherapy for breast cancer have improved patient survival. However, there continue to be nonresponders to conventional anticancer agents. Multidrug resistance (MDR) is caused by the expression of P-glycoprotein (P-gp) on the cell membrane. The expression of P-gp is encoded by MDR1 mRNA in tumors and is associated with clinical drug resistance. Since P-gp appears to be involved in both acquired and congenital MDR in human cancers, P-gp could be an important target for improving the efficacy of chemotherapy. Recently, we have focused on a therapeutic approach to reduce drug resistance in chemotherapy for breast cancer. Dofequidar fumarate (Dof) is a novel, orally active quinoline derivative that reverses multidrug resistance. In preclinical studies, the inhibition of doxorubicin-resistant cancer cell lines was observed in the presence of Dof + doxorubicin. We conducted clinical trials including Dof + cyclophosphamide (C), doxorubicin (A), and fluorouracil therapy (F) for patients with advanced or recurrent breast cancer. We compared the efficacy and tolerability of Dof + CAF with CAF alone. In this randomized, placebo-controlled trial, all patients were treated with six cycles of CAF therapy. Patients received Dof (900 mg p.o.) 30 min before doxorubicin. The primary endpoint was overall response rate (partial or complete response). In total, 221 patients were evaluable. The overall response rate was 42.6% for CAF alone versus 53.1% for Dof + CAF. Although the response rate improved by more than 10% with the combination of Dof + CAF, it was not statistically significant. Initially, we were expecting more than 20% improvement in the overall response rate. However, Dof significantly improved progression-free survival in patients who were premenopausal (P=0.046), who had received no prior therapy (P<0.01), or patients with advanced (stage IV) primary tumors (P=0.017). In addition, treatment with Dof did not affect the plasma concentration of doxorubicin in patients. These clinical studies indicate that Dof was well tolerated and displayed promising efficacy in patients who had not received prior therapy. The antiestrogens, tamoxifen, and toremifene, may moderate P-gp-related drug resistance in vitro. Toremifene demonstrated a synergistic effect in combination with paclitaxel on various human breast cancer cell lines. Furthermore, a synergistic effect was observed on a multidrug-resistant cell line. This synergistic effect was more potent when paclitaxel was combined with toremifene than with tamoxifen. Clinical benefits in some patients with recurrent breast cancer were reported. PMID- 16273363 TI - Translational studies for target-based drugs. AB - The biological background for the clinical and prognostic heterogeneity among tumors within the same histological subgroup is due to individual variations in the biology of tumors. The number of investigations looking at the application of novel technologies within the setting of clinical trials is increasing. The most promising way to improve cancer treatment is to build clinical research strategies on intricate biological evidence. New genomic technologies have been developed over recent years. These techniques are able to analyze thousands of genes and their expression profiles simultaneously. The purpose of this approach is to discover new cancer biomarkers, to improve diagnosis, predict clinical outcomes of disease and response to treatment, and to select new targets for novel agents with innovative mechanisms of action. Gene expression profiles are also used to assist in selecting biomarkers of pharmacodynamic effects of drugs in the clinical setting. Biomarker monitoring in surrogate tissues may allow researchers to assess "proof of principle" of new treatments. Clinical studies of biomarkers monitoring toxicity profiles have also been done. Such pharmacodynamic markers usually respond to treatment earlier than clinical response, and as such may be useful predictors of efficacy. Epidermal growth factor receptor (EGFR) mutation in lung cancer tissues is a strong predictive biomarker for EGFR targeted protein tyrosine kinase inhibitors. Monitoring of EGFR mutation has been broadly performed in retrospective and prospective clinical studies. However, global standardization for the assay system is essential for such molecular correlative studies. A more sensitive assay for EGFR mutation is now under evaluation for small biopsy samples. Microdissection for tumor samples is also useful for the sensitive detection of EGFR mutation. Novel approaches for the detection of EGFR mutation in other clinical samples such as cytology, pleural effusion and circulating tumor cells are ongoing. PMID- 16273364 TI - Chemotherapy for prostate cancer: implementing early systemic therapy to improve outcomes. AB - Prostate cancer remains a significant health concern for men in the USA as it is a leading cancer diagnosis and a cause of death. With the use of prostate specific antigen or screening, a stage migration has occurred with an increase in the number of men diagnosed with early-stage disease. The optimal primary management of these men is evolving, but despite adequate local treatment a significant percentage will develop either biochemical or clinical evidence of recurrent disease. Several criteria for risk stratification have been developed, thus, improving the ability to identify a high-risk population. Small studies have been reported demonstrating the feasibility of neoadjuvant or adjuvant chemotherapy in conjunction with either radiation or radical prostatectomy in this high-risk population, and large phase III studies are ongoing. With the advent of life-prolonging chemotherapy in the hormone-refractory setting, attention must now also be given to early-stage disease so as to develop multi modality approaches with the hope of increasing survival and ultimately providing a cure. PMID- 16273365 TI - Function of nuclear sex hormone receptors in gene regulation. AB - The development of reproductive organ tumors such as breast and prostate cancer often depends on the action of sex hormones. Nuclear sex hormone receptors are members of the nuclear hormone receptor superfamily and act as ligand-inducible transcription factors, controlling the expression of target genes. Nuclear receptors are considered to directly and indirectly interact with a number of nuclear co-regulatory complexes involved in chromatin remodeling and histone modification. Moreover, many intracellular signalings via cell membrane receptors are shown to modulate nuclear receptor-regulated transcription. We have shown that estrogen receptors (ER) associate with a number of nuclear complexes, one of which is a spliceosome complex. We recently found that this spliceosome complex interacts with phosphorylated ER by MAP kinase, generating a novel cross-talk of estrogen and growth factor signalings. We also observed that a dioxin receptor (AhR) is capable of associating with ER, resulting in modulation of ER transactivation function. From our findings we believe that development of estrogen-dependent breast cancer may be mediated through the other signaling pathways. To address the function of the androgen receptor (AR) in androgen dependent prostate cancer, we established a transgenic mouse line expressing a human AR mutant that is found in androgen-independent prostate cancer patients. The hAR mutant mice, generated through a Cre-loxP system, developed hyperplasia in the prostates. Hypersensitivity of AR mutants to antagonists and endogenous steroid hormones may potentiate hormone-dependency in prostate cancer development. PMID- 16273366 TI - The use of aromatase inhibitors in adjuvant therapy for early breast cancer. AB - Clinical evidence supporting the use of aromatase inhibitors (AIs) in adjuvant therapy for hormone-sensitive early breast cancer (EBC) has grown rapidly over the past few years and is reviewed in this article. The results of two studies the Arimidex, Tamoxifen, Alone or in Combination (ATAC) trial and the Breast International Group (BIG) 1-98 trial-support the use of AIs as primary adjuvant therapy for EBC, with significantly prolonged disease-free survival, time to recurrence, and time to distant recurrence for both anastrozole and letrozole over tamoxifen. Furthermore, anastrozole has an established beneficial risk:benefit ratio compared with tamoxifen with mature data over the full 5-year recommended treatment period. For women who have already received 2-3 years of tamoxifen, switching studies with anastrozole [the Italian Tamoxifen Anastrozole (ITA) trial and the Austrian Breast and Colorectal Cancer Study Group (ABCSG) 8/Arimidex-Nolvadex (ARNO 95) trial] and exemestane [the Intergroup Exemestane Study (IES)] have also shown that 5 years of primary adjuvant tamoxifen therapy is not optimal and that switching to an AI should be considered. Finally, for those women who have completed 5 years of tamoxifen, based on the results of the MA 17 trial, extended adjuvant treatment with letrozole should be considered. Although no sequencing data are available yet, current evidence suggests that an AI should be the adjuvant treatment of choice over tamoxifen, and anastrozole is the only AI with mature adjuvant data to date. PMID- 16273367 TI - Hormone treatment for prostate cancer: current issues and future directions. AB - Most prostate cancers are androgen-dependent and essentially respond to androgen ablation therapy. However, these tumors eventually become androgen-independent and progress despite androgen ablation. Since the androgen receptor (AR) sequence was determined, numerous studies have shown that AR plays a critical role in the development of androgen-refractory prostate cancer. Amplification of AR, mutations of AR, and deregulation of growth factors, cytokines and AR co activators, which could be classified as AR-dependent pathways, are frequently observed in this condition. There are other pathways, AR-independent pathways that bypass AR, which involve neuroendocrine differentiation of prostate cancer cells, deregulation of apoptotic genes and unknown mechanisms related to down regulation of AR. Androgen-refractory prostate cancers with the AR-dependent pathway could be treated by suppressing AR activity, whereas AR-independent tumors would require alternative management strategies. When more cell survival pathways are defined, improvement of patients' survival could be achieved by developing specific gene-targeting therapies that interfere with those pathways. PMID- 16273368 TI - The peculiar distribution of class I and class II aldolases in diatoms and in red algae. AB - Diatom plastids probably evolved by secondary endocytobiosis from a red alga that was up by a eukaryotic host cell. Apparently, this process increased the complexity of the intracellular distribution of metabolic enzymes. We identified genes encoding fructose-bisphosphate aldolases (FBA) in two centric (Odontella sinensis, Thalassiosira pseudonana) and one pennate (Phaeodactylum tricornutum) diatoms and found that four different aldolases are present in both groups: two plastid targeted class II enzymes (FBAC1 and FBAC2), one cytosolic class II (FBA3) and one cytosolic class I (FBA4) enzyme. The pennate Phaeodactylum possesses an additional plastidic class I enzyme (FBAC5). We verified the classification of the different aldolases in the diatoms by enzymatic characterization of isolated plastids and whole cell extracts. Interestingly, our results imply that in plastids of centric and pennate diatoms mainly either class I or class II aldolases are active. We also identified genes for both class I and class II aldolases in red algal EST databases, thus presenting a fascinating example of the reutilization and recompartmentalization of different aldolase isoenzymes during secondary endocytobiosis but as well demonstrating the limited use of metabolic enzymes as markers for the interpretation of phylogenetic histories in algae. PMID- 16273369 TI - Moc3, a novel Zn finger type protein involved in sexual development, ascus formation, and stress response of Schizosaccharomyces pombe. AB - The cAMP pathway in Schizosaccharomyces pombe is the major nutrient sensing pathway to initiate sexual development when opposite mating type cells exist. We identified moc1-moc4 as genes that overcome a partially sterile S. pombe strain due to an elevation of cAMP. When we compared the strength of inducing ability of sexual development in the same S. pombe strain, Moc1 had highest, Moc2 had lowest, and both Moc3 and Moc4 had intermediate effects. Moc1/Sds23 and Moc2/Ded1 are known to be a potential regulator of M-phase progression and an essential RNA helicase, respectively. While Moc4 was found to be identical with a Zn-finger protein Zfs1, Moc3 (SPAC821.07c) was a novel protein containing a Zn-finger (Zn(2)-Cys(6)) motif. Deletion mutant of the moc3 gene was constructed and its disruptant was found to be lower in mating efficiency and formed aberrant asci. In addition, unexpectedly, a moc3 disruptant was sensitive to CaCl(2) and DNA damaging agents such as MMS and UV. Those phenotypes were opposite to the phenotypes observed in a zfs1 disruptant, and quite different from the ones in a moc1 disruptant. Moc3 localized in the nucleus as observed for Zfs1. Moc3 bound with Moc4/Zfs1 weakly in the two hybrid system, but no other combination of Moc(s) bound each other in the same analysis. Thus, Moc3 is not only involved in sexual development, but also in ascus formation and DNA integrity in an independent manner with Moc1 and Moc2 in S. pombe. PMID- 16273370 TI - Heterologous expression of a pleiotropic drug resistance transporter from Phytophthora sojae in yeast transporter mutants. AB - A system for the expression of an ATP binding cassette (ABC) transporter from the soybean pathogen Phytophthora sojae is described. Pdr1, an ABC transporter with homology to the pleiotropic drug resistance (PDR) family of transporters, was cloned by primer walking from a P. sojae genomic library. Reverse transcriptase PCR assays showed that the transcript disappeared after encystment of zoospores and was not detected in hyphal germlings in dilute salts, in hyphae growing in liquid V8 media, or in tissue extracts from infected hypocotyls. BLAST analysis of Pdr1 against the P. sojae EST database also revealed that this gene was present only in zoospore libraries. Comparison of the number of hits to Pdr1 with that of a set of housekeeping genes revealed that Pdr1 was expressed at rates two to threefold higher than other transcripts. To test the hypothesis that Pdr1p functions as a broad substrate membrane transporter, Pdr1 was transformed into yeast mutants deficient in several drug resistance transporters. Yeast mutants transformed with Pdr1 possessed partial drug resistance against only 5 of 17 chemically distinct compounds. Thus, when expressed in yeast, this transporter has a significantly narrower substrate specificity in comparison to the yeast transporters, Pdr5p, Yorlp, and Snq2p. PMID- 16273371 TI - Intraductal papillary mucinous adenoma of the pancreas presenting with lobular panniculitis. PMID- 16273372 TI - The midterm success rates of tubularized incised plate urethroplasty in reoperative patients with distal or midpenile hypospadias. AB - To review the midterm results of tubularized incised plate (TIP) urethroplasty (Snodgrass method) in reoperative patients with distal or midpenile hypospadias. The results of TIP urethroplasty in 37 patients who had previously failed hypospadias repair were reviewed. Of the patients, 21 (56.8%) had coronal, 11 (29.7%) had subcoronal, and 5 (13.5%) had midpenile hypospadias. The mean age was 4.1 (2-16) years. Twenty-three patients had one operation and 14 patients had two operations previously. Of all the patients, 14 did not have a foreskin because of circumcision. The urethral plate had been disturbed in 6 patients, but there was not apparent scarring of the plate. Postoperative follow-up was 2.3 years with a range of 1.1-3.9 years. Genital examination, urethral calibration, and uroflowmetry were performed in control. Satisfaction of the families about the function and appearance of penis was also evaluated. Successful functional and cosmetic results were achieved in 29 patients (78.4%). All the families were happy with penile aesthetic appearance. The urethral plate seemed healthy at the operation in nine patients who had undergone TIP urethroplasty before and the outcomes were successful in eight of them. The operation was successful in 19/23 (82.6%) patients who had undergone one operation before and in 10/14 (71.4%) patients with two operations as well (P < 0.05). In addition, sufficient outcomes were obtained in also 11 of the 14 patients with circumcision. The success rate was higher in patients <5 years (P < 0.05). The rate was also higher during the recent period (2001-2003) since the experience we had increased (P < 0.05). TIP urethroplasty was unsatisfied in four of the six patients who had had disturbed urethral plate before and in five of eight patients who did not have sufficient amount of dartos tissue for flap to cover neourethra. Complication was observed in eight patients (21.6%): four had a pinpoint fistula, two had wound dehiscence, one had meatal stenosis, and one had mild meatal regression and a short neourethral stricture. All of these complications were repaired successfully at a later date. The mean hospital stay was 4.6 days. TIP urethroplasty provides good functional and cosmetic midterm outcomes in most of the reoperative patients with distal or midpenile hypospadias unless contraindicated by previous resection or gross scarring of the urethral plate. This procedure seems not to disturb the urethral plate and, therefore, it can be applied on reoperative patients who had undergone TIP urethroplasty before. It can also be used in a circumcised patient when there is a lack of foreskin. PMID- 16273373 TI - A complicated case of bilateral cystic nephroma in a 16-month-old boy. AB - We describe a case of a progressive bilateral cystic nephroma (BCN) in a child undergoing a multistaged surgical procedure. After partial resection of the left sided tumor, a progress occurred on that side and the left kidney had to be removed 10 weeks later. After 35 months a tumor progression was observed on the right side together with an ureteral obstruction leading to a decreased renal function. In a third operation a complete tumor resection on the right side was achieved through longitudinal partial nephrectomy, reconstruction of the renal pelvis, and reanastomosis of the reconstructed pelvis and ureter. The patient showed no evidence of the disease at 28 months of follow-up. The presented case provides an evidence that in BCN a tumor progress may occur after multistaged surgical approaches. A single-staged complete tumor resection with renal salvage techniques seems indicated. PMID- 16273374 TI - Delayed gastric emptying and malrotation in gastroesophageal reflux disease. PMID- 16273375 TI - A new autogenous graft choice in pelvic reconstructions: free vascularized rib (a case report). AB - Primary or secondary bone tumours are not uncommon in pelvic girdle. In some cases, after radical resection, there is a big bony defect where the prosthesis is not applicable; arthrodesis is the only choice for good functional results. In this instance, the major problem is to achieve the fusion. In this case report, we focused on easy harvesting and minimal time consumption with free vascularized rib graft to achieve the fusion between the resected segments. Two year follow up showed fusion with good functional result. PMID- 16273376 TI - Management of the bilateral congential dislocation of the hip and knee: a case report. AB - Congenital dislocation of the knee (CDK) is a rare disease and may be associated with other congenital and musculoskeletal disorders. We report our result in the treatment of a patient with CDK and DDH that treated with serial casting and Pavlik harness. Early diagnosis of the CDK is very important non operative treatment usually provides more stabile, greater range of motion and much more quadriceps strength than the surgical treatment. Early reduction of dislocation prevents formation of the knee contracture. PMID- 16273377 TI - Correction of severe post-traumatic deformities in the distal femur by distraction osteogenesis using Taylor Spatial Frame: a case report. AB - A case of deformity and shortening after post-traumatic growth arrest treated using the Taylor Spatial Frame (Smith & Nephew, Tennessee, USA) is presented. This is the first report showing the application of the frame for post-traumatic deformity in the distal femur, and successful outcomes promise utilization of the frame even for correction of severe deformity in the distal femur. PMID- 16273378 TI - Modified Steindler procedure for the treatment of brachial plexus injuries. AB - A retrospective follow-up study was completed on ten patients who suffered from a brachial plexus injury that was treated with a modified Steindler procedure. The mean postoperative period was 6.8 years. The postoperative elbow joint range of motion was -42 degrees of extension (range -5 degrees to -65 degrees ) and 107 degrees of flexion (range 90 degrees -130 degrees ). Manual muscle testing showed grade 4 or 5 in eight patients and grade 3 in two patients. In the subjective assessment, the patients scored 20 out of 30 points and were able to perform almost all activities with the exception of shoulder elevation. Innervation of the musculocutaneous nerve was evaluated by electromyography and no correlation was seen between preoperative and postoperative amplitude of the biceps brachii by electromyogram. Based on these results, we concluded that a modified Steindler procedure is useful for reconstruction of upper brachial plexus injuries, and recovery of the biceps brachii was difficult to predict by an electromyogram. PMID- 16273379 TI - Joint kinematics and in situ forces after single bundle PCL reconstruction: a graft placed at the center of the femoral attachment does not restore normal posterior laxity. AB - INTRODUCTION: Femoral tunnel placement has a great influence on the clinical outcome after PCL reconstruction. MATERIALS AND METHODS: Using a robotic/universal force moment sensor (UFS) testing system, we examined joint kinematics and in situ forces of human knees following soft-tissue single bundle PCL reconstruction fixed at the center of the femoral attachment. RESULTS: Posterior tibial translation significantly increased at all flexion angles after transsection of the posterior cruciate ligament (p<0.05). PCL reconstruction resulted in significantly less posterior tibial translation at all flexion angles when compared to the PCL deficient knee (p<0.05). The differences in the in situ force between the intact ligament and the reconstructed graft were statistical significant (p<0.05). CONCLUSION: Single bundle PCL reconstruction with a soft tissue graft fixed at the center of the femoral attachment is able to reduce the posterior tibial translation significantly. However, it cannot restore kinematics of the intact knee and in situ forces of the intact PCL. PMID- 16273380 TI - The anterior approach for the treatment of posterior osteochondral lesions of the talus: comparison of different surgical techniques. AB - INTRODUCTION: Most of all osteochondral talar lesions are located in the middle and posterior area of the talar surface. Malleolar osteotomy is often used to access the defect but may be associated with malunion or secondary osteoarthritis. We present an alternative approach to the talus with temporary removal and replacement of a tibial bone block and compare it with other anterior approaches described in the literature. PATIENTS AND METHODS: Thirteen patients (5 males, 8 females) with an average age of 27.2 years and an osteochondral talar lesion were included in our study. All patients were previously operated on the same ankle. Ten lesions were caused by a sports injury. The average follow up was 45 months. The patients were evaluated before and after surgery using the ankle and hindfood score (AOFAS). For the analyses baseline clinical data were compared with follow up data using the Wilcoxon test. RESULTS: The overall improvement between the preoperative and postoperative AOFAS scores was an average of 34.9 points (P=0.0002). No complications occurred at the site of the tibial bone block and the donor site at the talus. There were no patients with recurrence or an ankle osteoarthrosis in the follow up period. CONCLUSION: The removal of a tibial bone block and its subsequent replacement is a useful technique to access osteochondral talar lesions for osteochondral transplantation for which arthroscopic interventions have failed. The results are comparable to other anterior approaches described in the literature. PMID- 16273381 TI - Radiation risk estimates after radiotherapy: application of the organ equivalent dose concept to plateau dose-response relationships. AB - Estimates of secondary cancer risk after radiotherapy are becoming more important for comparative treatment planning. Modern treatment planning systems provide accurate three-dimensional (3D) dose distributions for each individual patient. The dose distributions can be converted into organ equivalent doses to describe radiation-induced cancer after radiotherapy (OED(rad-ther)) in the irradiated organs. The OED(rad-ther) concept assumes that any two dose distributions in an organ are equivalent if they cause the same radiation-induced cancer risk. In this work, this concept is applied to dose-response relationships, which are leveling off at high dose. The organ-dependent operational parameter of this dose response relationship was estimated by analyzing secondary cancer incidence data of patients with Hodgkin's disease. The dose distributions of a typical radiotherapy treatment plan for treating Hodgkin's disease was reconstructed. Dose distributions were calculated in individual organs from which cancer incidence data were available. The model parameter was obtained by comparing dose and cancer incidence rates for the individual organs. PMID- 16273382 TI - Influence of mitotic delay on the results of biological dosimetry for high doses of ionizing radiation. AB - The purpose of this study was to systematically investigate how high doses of sparsely and densely ionizing radiations influence the proliferation time of lymphocytes in short-term cultures and, consequently, the observed frequencies of dicentric and centric ring chromosomes. Peripheral blood samples from five volunteers were irradiated with high doses of 200 kV X-rays and with neutrons with a mean energy of or=2.1 MeV. First division metaphase cells were collected after different culture times of 48, 56, and 72 h and dicentrics, centric ring chromosomes, and acentric fragments were determined. The data hint at considerable mitotic delay. The main increase in the number of chromosome aberrations occurred between 48 and 72 h after an X-ray exposure and between 56 and 72 h after neutron exposure. When the data were used for a calibration of aberration frequency versus dose, subsequent dose estimations resulted, however, in comparable values. Thus, in spite of the influence of mitotic delay on observable chromosome aberrations, at least for the radiation types investigated here, a culture time of 48 h is acceptable for biological dosimetry. PMID- 16273383 TI - Cell death in trichomonads: new insights. AB - Tritrichomonas foetus is an amitochondriate parasite that possesses hydrogenosomes, unusual anerobic energy-producing organelles. In these organisms the "mitochondrial cell death machinery" is supposed to be absent, and the mechanisms that lead to cell demise remain to be elucidated. The presence of a cell death program in trichomonads has already been reported, suggesting the existence of a caspase-like execution pathway in such organisms. Here we demonstrate the alterations provoked by the fungicide griseofulvin and raise the possibility that other cell death pathways may exist in T. foetus. Dramatic changes in trichomonads morphology are presented after griseofulvin treatment, such as intense plasma membrane and nuclear envelope blebbing, nucleus fragmentation, and an abnormal number of oversized vacuoles. One important finding was the exposition of phosphatidylserine (PS) in the outer leaflet of the plasma membrane in cells after drug treatment, and also the presence of a high amount of misshapen flagella and tubulin precipitates as vacuolar contents, suggesting an autophagic process of abnormal cellular elements. Interestingly, immunoreactivity for activated caspase-3 was not detected during griseofulvin treatment, a finding distinct from the observed when this cell was treated with H(2)O(2). The possibility of the existence of different pathways to cell death in trichomonads is discussed. PMID- 16273384 TI - A novel population of neuronal cells expressing the olfactory marker protein (OMP) in the anterior/dorsal region of the nasal cavity. AB - The olfactory marker protein (OMP) is expressed in mature chemosensory neurons in the nasal neuroepithelium. Here, we report the identification of a novel population of OMP-expressing neurons located bilaterally in the anterior/dorsal region of each nasal cavity at the septum. These cells are clearly separated from the regio olfactoria, harboring the olfactory sensory neurons. During mouse development, the arrangement of the anterior OMP-cells undergoes considerable change. They appear at about stage E13 and are localized in the nasal epithelium during early stages; by epithelial budding, ganglion-shaped clusters are formed in the mesenchyme during the perinatal phase, and a filiform layer directly underneath the nasal epithelium is established in adults. The anterior OMP-cells extend long axonal processes which form bundles and project towards the brain. The data suggest that the newly discovered group of OMP-cells in the anterior region of the nasal cavity may serve a distinct sensory function. PMID- 16273385 TI - How to find decision makers in neural networks. AB - Nervous systems often face the problem of classifying stimuli and making decisions based on these classifications. The neurons involved in these tasks can be characterized as sensory or motor, according to their correlation with sensory stimulus or motor response. In this study we define a third class of neurons responsible for making perceptual decisions. Our mathematical formalism enables the weighting of neuronal units according to their contribution to decision making, thus narrowing the field for more detailed studies of underlying mechanisms. We develop two definitions of a contribution to decision making. The first definition states that decision making activity can be found at the points of emergence for behavioral correlations in the system. The second definition involves the study of propagation of noise in the network. The latter definition is shown to be equivalent to the first one in the cases when they can be compared. Our results suggest a new approach to analyzing decision making networks. PMID- 16273387 TI - Large-scale morphometric analysis of neuroanatomy and neuropathology. AB - Brain imaging research with MRI spans a wide area, covering both structure and function, and ranging from basic research through clinical research to drug design and clinical trials. In recent years there has been a trend towards the collection of very large MRI databases which can allow for the detection of very small group-dependent effects. However, the logistical challenges of analysing such large datasets presents new challenges. This paper describes the "pipeline" framework developed at the Montreal Neurological Institute for the fully automated morphometric analysis of large brain imaging databases. The potential use of these techniques is illustrated by examples of their applications in multiple sclerosis, Alzheimer's disease, and pediatric development. PMID- 16273386 TI - An adiponectin receptor, T-cadherin, was selectively expressed in intratumoral capillary endothelial cells in hepatocellular carcinoma: possible cross talk between T-cadherin and FGF-2 pathways. AB - T-cadherin is a unique receptor of adiponectin, which plays a critical role in various angiogenesis. In the present study, T-cadherin expression in tumor vessels of hepatocellular carcinoma (HCC) and, subsequently, the molecular mechanism, which induced T-cadherin expression in sinusoidal endothelial cells were investigated. Sinusoidal endothelium in nontumorous liver, chronic hepatitis, or liver cirrhosis expressed little or no T-cadherin. By contrast, T cadherin was found in intratumoral capillary endothelial cells of 34 out of 63 HCC specimens. In positive cases, focal T-cadherin expression was found in well differentiated HCC, whereas diffuse and intense T-cadherin expression was observed in poorly differentiated HCC specimens. T-cadherin was much expressed in intratumoral capillary endothelial cells in a less differentiated HCC region than that in a well-differentiated region in five specimens, in which various differentiated HCC components were coexistent. In a double-cell chamber assay, fibroblast growth factor-2 appeared to have a critical role to induce T-cadherin in cultured liver sinusoidal endothelial cells. The present finding indicated that T-cadherin was selectively expressed in intratumoral capillary endothelial cells of many HCCs, increasingly expressed as tumor progression, and T-cadherin may have a positive role in angiogenesis of HCC. In addition, cross talk between the signal pathways mediated by fibroblast growth factor-2 and adiponectin was suggested. PMID- 16273388 TI - Significant microsynteny with new evolutionary highlights is detected between Arabidopsis and legume model plants despite the lack of macrosynteny. AB - The increased amount of data produced by large genome sequencing projects allows scientists to carry out important syntenic studies to a great extent. Detailed genetic maps and entirely or partially sequenced genomes are compared, and macro- and microsyntenic relations can be determined for different species. In our study, the syntenic relationships between key legume plants and two model plants, Arabidopsis thaliana and Populus trichocarpa were investigated. The comparison of the map position of 172 gene-based Medicago sativa markers to the organization of homologous A. thaliana genes could not identify any sign of macrosynteny between the two genomes. A 276 kb long section of chromosome 5 of the model legume Medicago truncatula was used to investigate potential microsynteny with the other legume Lotus japonicus, as well as with Arabidopsis and Populus. Besides the overall correlation found between the legume plants, the comparison revealed several microsyntenic regions in the two more distant plants with significant resemblance. Despite the large phylogenetic distance, clear microsyntenic regions between Medicago and Arabidopsis or Populus were detected unraveling new intragenomic evolutionary relations in Arabidopsis. PMID- 16273389 TI - Localization of a novel locus for alopecia with mental retardation syndrome to chromosome 3q26.33-q27.3. AB - Alopecia with mental retardation syndrome is a rare autosomal recessive disorder characterized clinically by total or partial alopecia and mental retardation. In an effort to understand the molecular bases of this form of alopecia syndrome, large Pakistani consanguineous kindred with multiple affected individuals has been ascertained from a remote region in Pakistan. Genome wide scan mapped the disease locus on chromosome 3q26.33-q27.3. A maximum two-point LOD score of 3.05 (theta = 0.0) was obtained at marker D3S3583. Maximum multipoint LOD score exceeding 5.0, obtained with several markers, supported the linkage. Recombination events observed in affected individuals localized the disease locus between markers D3S1232 and D3S2436, spanning 11.49-cM region on chromosome 3q26.33-q27.3. Sequence analysis of a candidate gene ETS variant gene 5 from DNA samples of two affected individuals of the family revealed no mutation. PMID- 16273390 TI - Admixture-matched case-control study: a practical approach for genetic association studies in admixed populations. AB - Case-control genetic association studies in admixed populations are known to be susceptible to genetic confounding due to population stratification. The transmission/disequilibrium test (TDT) approach can avoid this problem. However, the TDT is expensive and impractical for late-onset diseases. Case-control study designs, in which, cases and controls are matched by admixture, can be an appealing and a suitable alternative for genetic association studies in admixed populations. In this study, we applied this matching strategy when recruiting our African American participants in the Study of African American, Asthma, Genes and Environments. Group admixture in this cohort consists of 83% African ancestry and 17% European ancestry, which was consistent with reports from other studies. By carrying out several complementary analyses, our results show that there is a substructure in the cohort, but that the admixture distributions are almost identical in cases and controls, and also in cases only. We performed association tests for asthma-related traits with ancestry, and only found that FEV(1), a measure for baseline pulmonary function, was associated with ancestry after adjusting for socio-economic and environmental risk factors (P=0.01). We did not observe an excess of type I error rate in our association tests for ancestry informative markers and asthma-related phenotypes when ancestry was not adjusted in the analyses. Furthermore, using the association tests between genetic variants in a known asthma candidate gene, beta(2) adrenergic receptor (beta(2)AR) and DeltaFEF(25-75), an asthma-related phenotype, as an example, we demonstrated population stratification was not a confounder in our genetic association. Our present work demonstrates that admixture-matched case-control strategies can efficiently control population stratification confounding in admixed populations. PMID- 16273391 TI - Genome-wide linkage of febrile seizures and epilepsy to the FEB4 locus at 5q14.3 q23.1 and no MASS1 mutation. AB - Febrile seizures (FS) represent the most common seizure disorder in childhood and contribution of a genetic predisposition has been clearly proven. In some families FS is associated with a wide variety of afebrile seizures. Generalized epilepsy with febrile seizures plus (GEFS+) is a familial epilepsy syndrome with a spectrum of phenotypes including FS, atypical febrile seizures (FS+) and afebrile generalized and partial seizures. Mutations in the genes SCN1B, SCN1A and GABRG2 were identified in GEFS+ families. GEFS+ is genetically heterogeneous and mutations in these three genes were detected in only a minority of the families. We performed a 10 cM density genome-wide scan in a multigenerational family with febrile seizures and epilepsy and obtained a maximal multipoint LOD score of 3.12 with markers on chromosome 5q14.3-q23.1. Fine mapping and segregation analysis defined a genetic interval of approximately 33 cM between D5S2103 and D5S1975. This candidate region overlapped with a previously reported locus for febrile seizures (FEB4) in the Japanese population, in which MASS1 was proposed as disease gene. Mutation analysis of the exons and exon-intron boundaries of MASS1 in our family did not reveal a disease causing mutation. Our linkage data confirm for the first time that a locus on chromosome 5q14-q23 plays a role in idiopathic epilepsies. However, our mutation data is negative and do not support a role for MASS1 suggesting that another gene within or near the FEB4 locus might exist. PMID- 16273392 TI - Ectomycorrhizal symbiosis in vitro between Tricholoma matsutake and Pinus densiflora seedlings that resembles naturally occurring 'shiro'. AB - We established an in vitro ectomycorrhizal symbiosis between Tricholoma matsutake and Pinus densiflora. Mycorrhiza formed in a substrate of Modified Norkrans' C medium and granite-based soil had features similar to those observed previously only in naturally occurring mycorrhizal system called 'shiro,' and promoted the growth of plants with smaller root/shoot ratios. The in vitro formation of 'shiro' is essential for the development of T. matsutake system to produce mushrooms and is useful for the propagation and plantation of the mycorrhizal seedlings. PMID- 16273393 TI - Amt/MEP/Rh proteins conduct ammonia. AB - The structure determination of the ammonium transport protein AmtB from Escherichia coli strongly indicates that the members of the ubiquitous ammonium transporter/methylamine permease/Rhesus (Amt/MEP/Rh) protein family are ammonia conducting channels rather than ammonium ion transporters. The most conserved part of these proteins, apart from the common overall structure with 11 transmembrane helices, is the pore lined by hydrophobic side chains except for two highly conserved histidine residues. A high-affinity ion-binding site specific for ammonium is present at the extracellular pore entry of the Amt/MEP proteins. It is proposed to play an important role in enhancing net transport at very low external ammonium concentrations and to provide discrimination against water. The site is not conserved in the animal Rhesus proteins which are implicated in ammonium homeostasis and saturate at millimolar ammonium concentrations. Many aspects of the biological function of these ammonia channels are still poorly understood and further studies in cellular systems are needed. Likewise, studies with purified, reconstituted Amt/MEP/Rh proteins will be needed to resolve open mechanistic questions and gain a more quantitative understanding of the conduction mechanism in general and for different subfamily representatives. PMID- 16273394 TI - Excitability of the pathways mediating the startle reaction before execution of a voluntary movement. AB - Studies with transcranial electrical or magnetic stimulation have shown a progressive increase of motor cortex excitability beginning at about 80 ms before the onset of electromyographic (EMG) activity in a voluntary movement. We studied whether a similar increase in excitability occurs in subcortical motor tracts before execution of a ballistic movement. In ten healthy volunteers, we examined the effects of a startling auditory stimulus (SAS) applied at various intervals following the imperative signal (IS) in a reaction time task experiment. We hypothesized that, if the excitability of the reticulospinal tract increases before onset of muscle activity, there would be a corresponding change in the size of the startle response elicited either in muscles responding to the SAS, the orbicularis oculi (OOc) and sternocleidomastoid (SCM), or in the agonist muscle for the reaction. A SAS was applied at intervals of 0, 20, 40, 60, 80, and 100 ms after IS in a reaction time task paradigm in which subjects had to perform a ballistic movement of wrist flexion. We measured the size of the EMG bursts recorded in the OOc and the SCM, as well as the size of the first EMG burst in the wrist flexors (WF). Comparisons were done with data obtained in baseline trials, in which unexpected SAS of the same intensity were delivered without preparation, and control trials, in which subjects performed the same motor task with no SAS. The size of the averaged OOc, SCM, and WF were larger in trials with SAS than in baseline or control trials, with significant differences in the SCM and in the WF (p<0.05). However, there were no effects of time interval on the size of OOc, SCM, or WF (p>0.05). These results indicate that, in the execution of a ballistic movement, the excitability of the subcortical motor tracts activated by a SAS is similarly enhanced at the time of the IS and 100 ms afterwards, just before the onset of EMG activity. We conclude that, in contrast with the reported progressive increase of excitability in the corticospinal tract, the excitability of the tracts activated by a SAS do not change between the IS and the onset of EMG activity. PMID- 16273396 TI - Lower extremity control and dynamics during backward angular impulse generation in backward translating tasks. AB - Observation of complex whole-body movements suggests that the nervous system coordinates multiple operational subsystems using some type of hierarchical control. When comparing two backward translating tasks performed with and without backward angular impulse, we have learned that task-specific modifications in trunk-leg coordination contribute to the regulation of total-body center of mass (CoM) position relative to the reaction force (RF). In this study, we hypothesized that task-specific differences in trunk-leg coordination would affect the control of the lower extremity joints during the impulse-generation phase of the tasks. Eight highly skilled performers executed a series of backward translating jumps with and without backward rotation (back somersault and back timer, respectively). Sagittal plane kinematics, RFs and electromyograms of lower extremity muscles were acquired during the take-off phase of both tasks. Lower extremity joint kinetics was calculated using inverse dynamics. The results indicate that between-task differences in the relative angles between the lower extremity segments and the net joint forces/RF contributed to significant reductions in knee-extensor net joint moments and increases in hip-extensor net joint moments during the push interval of the back somersault as compared to the back timer. Between-task differences in backward trunk angular velocity also contributed to the re-distribution of work done by the lower extremity net joint moments. Between-task differences in lower extremity joint kinetics were associated with synergistic activation of the bi-articular muscles crossing the knee and hip. These results indicated that task-specific control of CoM relative to the RF in order to regulate the backward angular-impulse-involved modification in the control and dynamics of the knee and hip joints. These results indicate that between-task differences in the control objectives at the total-body level (position of CoM relative to the RF) alters the control and dynamics of the multi joint lower extremity subsystem. PMID- 16273395 TI - Differential effects of subthalamic nucleus stimulation in advanced Parkinson disease on reaction time performance. AB - The aim of the present study was to assess the effect of bilateral subthalamic nucleus (STN) stimulation and dopaminergic medication on speed of mental processing and motor function. Thirty-nine patients suffering from advanced Parkinson disease (PD) were operated on. Motor function and reaction time (RT) performance [simple RT (SRT) and complex RT (CRT)] were evaluated under four experimental conditions with stimulation (stim) and medication (med) on and off: stim-on/med-on, stim-on/med-off, stim-off/med-off and stim-off/med-on. In the last condition, the patients received either low medication (usual dose) or high medication (suprathreshold dose). STN stimulation improved the motor performance in the SRT and CRT tasks. Furthermore, STN deep brain stimulation (DBS) also improved response preparation as shown by the significant improvement of the RT performance in the SRT task. This effect of STN DBS on the RT performance in the SRT task was greater as compared with the CRT task. This is due to the more complex information processing that is required in the CRT task as compared to the SRT task. These data suggest that treatment of STN hyperactivity by DBS improves motor function, confirming earlier reports, but has a differential effect on cognitive functions. The STN seems to be an important modulator of cognitive processing and STN DBS can differentially affect motor and associative circuits. PMID- 16273397 TI - Our eyes deviate away from a location where a distractor is expected to appear. AB - Previous research has shown that in order to make an accurate saccade to a target object, nearby distractor objects need to be inhibited. The extent to which saccade trajectories deviate away from a distractor is often considered to be an index of the strength of inhibition. The present study shows that the mere expectation that a distractor will appear at a specific location is enough to generate saccade deviations away from this location. This suggests that higher order cognitive processes such as top-down expectancy interact with low-level structures involved in eye movement control. The results will be discussed in the light of current theories of target selection and possible neurophysiological correlates. PMID- 16273398 TI - The use of quantitative oculometry in the assessment of Huntington's disease. AB - Huntington's disease (HD) is a neurodegenerative disorder characterised by progressive motor, cognitive and psychiatric symptoms. Objective measurement of disease severity is of increasing importance for detecting symptomatic disease, as well as monitoring disease progression and the response to novel therapeutic interventions. Using a newly-developed infra-red scleral oculometer, we measured saccadic latencies and durations in HD patients exhibiting a broad range of symptoms (n=24) and control subjects of comparable ages (n=20) to see whether these parameters might reflect the presence or severity of HD. Latency distributions were characterised by creating reciprobit plots for each subject, whilst parametric statistics were applied to durations. Compared with the control group, we found the HD group had a significantly increased median latency, and early saccades were more prominent. In addition, HD patients exhibited an increased saccadic duration and variability of duration. Using Bayesian (likelihood) analysis, we obtained saccadic support values for the presence of clinical HD, which correlated with the motor Unified Huntington's Disease Rating Scale (UHDRS) score in these patients. A sensitivity/specificity analysis of all 44 participants showed that the use of this multivariate support measure was highly successful in predicting HD status, correctly diagnosing 75% of the HD patients, and (95%) of the controls; with a different criterion, these figures were 96 and 15%. Furthermore, we correctly predicted absence of disease in two additional subjects subsequently confirmed to be genetically unaffected. This strongly suggests that multivariate support values derived from saccadic parameters may provide an objective, quantitative biomarker of HD, especially the degree of motor impairment. However, larger longitudinal studies are required to determine whether they can reliably detect the earliest presymptomatic disease, or faithfully reflect disease progression. PMID- 16273399 TI - Attentional demands associated with the use of a light fingertip touch for postural control during quiet standing. AB - The purpose of the present experiment was to investigate whether and how using a light fingertip touch for postural control during quiet standing requires additional attentional demands. Nine young healthy university students were asked to respond as rapidly as possible to an unpredictable auditory stimulus while maintaining stable seated and upright postures in three sensory conditions: vision, no-vision and no-vision/touch. Touch condition involved a gentle light touch with the right index finger on a nearby surface at waist height. Center of foot pressure (CoP) displacements were recorded using a force platform. Reaction times (RTs) values were used as an index of the attentional demand necessary for calibrating the postural system. Results showed decreased CoP displacements in both the vision and no-vision/touch conditions relative to the no-vision condition. More interestingly, a longer RT in the no-vision/touch than in the vision and no-vision conditions was observed. The present findings suggest that the ability to use a light fingertip touch as a source of sensory information to improve postural control during quiet standing is attention demanding. PMID- 16273400 TI - A role for terrain slope in orienting hippocampal place fields. AB - The three-dimensional topography of the environment is a potentially important source of orienting information for animals, but little is known about how such features affect either navigational behaviour or the neural representation of place. One component of the neural place representation comprises the hippocampal place cells, which show location-specific firing that can be oriented by directional cues in the environment. The present study investigated whether a simple topographical feature, terrain slope, could provide such orienting information to place cells. Place cells were recorded as rats explored a tilted (30 degrees) square box located in the centre of a dark, curtained and visually symmetrical circular enclosure. The orientation of the tilted surface was varied, first in conjunction with that of a visible cue card (to stabilise the system) and then in the absence of the cue card, when the slope of the box was the only remaining stable polarising cue in the environment. In the latter condition, place fields continued to be reliably oriented by the slope. Thus, terrain slope provides sufficient orienting information to set and probably maintain the orientation of the hippocampal place system. This may explain previous behavioural observations that spatial orientation is improved when slope information is available. PMID- 16273401 TI - A single re-implanted ventral root exerts neurotropic effects over multiple spinal cord segments in the adult rat. AB - Spinal cord injuries, particularly traumatic injuries to the conus medullaris and cauda equina, are typically complex and involve multiple segmental levels. Implantation of avulsed ventral roots into the spinal cord as a repair strategy has been shown to be neuroprotective and promote axonal regeneration by spinal cord neurons into an implanted root. However, it is not well known over what distance in the spinal cord an implanted ventral root can exert its neurotropic effect. Here, we investigated whether an avulsed L6 ventral root acutely implanted into the rat spinal cord after a four level (L5-S2) unilateral ventral root avulsion injury may exert neurotropic effects on autonomic and motor neurons over multiple spinal cord segments at 6 weeks postoperatively. Using retrograde labeling techniques and stereological quantification methods, we demonstrate that autonomic and motor neurons from all four lesioned spinal cord segments, spanning more than an 8 mm rostro-caudal distance, reinnervated the one implanted root. The rostro-caudal distribution suggested a gradient of neurotropism, where the axotomized neurons closest to the implanted site had the highest probability of root reinnervation. These results suggest that implantation of a single ventral root may provide neurotropic effects to injured neurons at the site of lesion as well as in the adjacent spinal cord segments. Our findings may be of translational research interest for the development of surgical repair strategies after multi-level conus medullaris and cauda equina injuries, in which fewer ventral roots than spinal cord segments may be available for implantation. PMID- 16273402 TI - Smooth pursuit rather than visual signals mediate short-term adaptation of the cervico-ocular reflex in humans. AB - Cervical stimulation in the horizontal plane induces small and variable eye movements in normal human adults [cervico-ocular reflex (COR)]. In patients with bilateral vestibular loss, the slow COR component increases in amplitude and is thought to contribute to gaze stabilization during head movements, as it is directed opposite to head-on-trunk excursions. The procedures underlying COR slow phase gain enhancement in the compensatory direction remain unclear. We studied the horizontal COR during passive trunk oscillations of +/-16 degrees under the stationary head at 0.1 Hz in ten normal subjects, aged 24-30 years, before and immediately after the application of an adaptation procedure engaging various combinations of sinusoidal neck-proprioceptive, pursuit and retinal slip signals. The duration of this adaptation period was 40 min. A significant gain increase and phase modulation in the compensatory direction were observed in four out of eight subjects after exposing them to neck-proprioceptive stimulation, while pursuing a spot moving in-phase with their trunk. In contrast, staring at the rotating optokinetic pattern or fixating at a stationary spot, while being subjected to combined cervical and optokinetic stimulation, failed to result in any significant modification of the subjects' COR gain and phase. Conceivably, the contribution of the pursuit system was greatly reduced in the paradigm using optokinetic stimulation, while full engagement of retinal slip signals, in the absence of any pursuit contribution, was obtained in the latter adaptation paradigm. These results indicate that motor responses of target tracking rather than simply sensory signals of retinal slip may represent the 'error signal' modifying the COR in humans. PMID- 16273403 TI - The attentional mechanism of temporal orienting: determinants and attributes. AB - A review of traditional research on preparation and foreperiod has identified strategic (endogenous) and automatic (exogenous) factors probably involved in endogenous temporal-orienting experiments, such as the type of task, the way by which temporal expectancy is manipulated, the probability of target occurrence and automatic sequential effects, yet their combined impact had not been investigated. These factors were manipulated within the same temporal-orienting procedure, in which a temporal cue indicated that the target could appear after an interval of either 400 or 1,400 ms. We observed faster reaction times for validly versus invalidly cued targets, that is, endogenous temporal-orienting effects. The main results were that the probability of target occurrence (catch trial proportion) modulated temporal orienting, such that the attentional effects at the short interval were independent of catch trials, whereas at the long interval the effects were only observed when catch trials were present. In contrast, the interval duration of the previous trial (i.e., exogenous sequential effects) did not influence endogenous temporal orienting. A flexible and endogenous mechanism of attentional orienting in time can account for these results. Despite the contribution of other factors, the use of predictive temporal cues was sufficient to yield attentional facilitation based on temporal expectancy. PMID- 16273404 TI - Behavioral and neurophysiological effects of delayed training following a small ischemic infarct in primary motor cortex of squirrel monkeys. AB - A focal injury within the cerebral cortex results in functional reorganization within the spared cortex through time-dependent metabolic and physiological reactions. Physiological changes are also associated with specific post-injury behavioral experiences. Knowing how these factors interact can be beneficial in planning rehabilitative intervention after a stroke. The purpose of this study was to assess the functional impact of delaying the rehabilitative behavioral experience upon movement representations within the primary motor cortex (M1) in an established nonhuman primate, ischemic infarct model. Five adult squirrel monkeys were trained on a motor-skill task prior to and 1 month after an experimental ischemic infarct was induced in M1. Movement representations of the hand were derived within M1 using standard electrophysiological procedures prior to the infarct and again one and two months after the infarct. The results of this study show that even though recovery of motor skills was similar to that of a previous study in squirrel monkeys after early training, unlike early training, delayed training did not result in maintenance of the spared hand representation within the M1 peri-infarct hand area. Instead, delaying training resulted in a large decrease in spared hand representation during the spontaneous recovery period that persisted following the delayed training. In addition, delayed training resulted in an increase of simultaneously evoked movements that are typically independent. These results indicate that post-injury behavioral experience, such as motor skill training, may modulate peri-infarct cortical plasticity in different ways in the acute versus chronic stages following stroke. PMID- 16273405 TI - Metabotropic glutamate receptor antagonists but not NMDA antagonists affect conditioned taste aversion acquisition in the parabrachial nucleus of rats. AB - The effect of glutamate receptor antagonists on conditioned taste aversion (CTA) was studied in rats. The association of the short-term memory of a gustatory conditioned stimulus (CS) with visceral malaise (unconditioned stimulus, US) in the CTA paradigm takes place in the parabrachial nuclei (PBN) of the brainstem. The first direct evidence of participation of glutamatergic neurotransmission in the PBN during CTA demonstrated that the extracellular level of glutamate rises during saccharin drinking (Bielavska et al. in Brain Res 887:413-417, 2000). Our results show an effect of microdialysis administration of selective GluR antagonists into the PBN on the formation of CTA engram. We used four glutamate receptor (GluR) antagonists of different types (D-AP5, MK-801 as antagonists of ionotropic GluR and L-AP3, MSPG as antagonists of metabotropic GluR). The disruptive effect of MK-801 on CTA formation in the PBN is concentration dependent, with the greatest inhibition under the higher concentrations eliciting significant disruption. The application of D-AP5 (0.1, 1, 5 mM) did not elicit a statistically significant blockade of CTA acquisition. This indicates that the association of the US-CS in the PBN is not dependent on NMDA receptors. On the contrary, application of L-AP3 (0.1, 1, 5 mM) blocked the CS-US association. PMID- 16273408 TI - Clinical experience with decitabine in North American patients with myelodysplastic syndrome. AB - Recent evidence demonstrates that epigenetic silencing of genes is associated with myelodysplasia and that a worse prognosis may be correlated with hypermethylation of certain genes, such as the cyclin-dependent kinase inhibitor p15. 5-Aza-2'-deoxycytidine (decitabine, DAC) is a nucleoside analog, which, at low doses, acts as a hypomethylating agent and is fivefold to tenfold more active than 5-azacytidine (azacitidine, Vidaza)--currently the only approved drug for treatment of myelodysplastic syndrome (MDS). Clinical studies have demonstrated that decitabine has activity in patients with MDS. Preliminary results of a phase III multicenter North American trial comparing low-dose decitabine to supportive care verified that therapy with decitabine resulted in higher response rates, improved quality of life, and prolonged time to leukemic transformation and/or death. However, further elucidation of its mechanism of action is required, as clinical response to decitabine does not correlate with demethylation of the p15 gene promoter or the repetitive DNA element LINE. Decitabine appears to upregulate both hypermethylated and nonmethylated genes. Ongoing studies aim to determine the optimal dose, schedule, and route of administration of decitabine, and to evaluate whether efficacy can be improved by using it in combination with other agents, such as histone deacetylase inhibitors. PMID- 16273407 TI - Are somatosensory saccades voluntary or reflexive? AB - The present study examines whether the distinction between voluntary (endogenous) and reflexive (stimulus-elicited) saccades made in the visual modality can be applied to the somatosensory modality. The behavioural characteristics of putative reflexive pro-saccades and voluntary anti-saccades made to visual and somatosensory stimuli were examined. Both visual and somatosensory pro-saccades had much shorter latency than voluntary anti-saccades made in the direction opposite to a peripheral stimulus. Furthermore, erroneous pro-saccades were made towards both visual and somatosensory stimuli on approximately 11-13% of anti saccade trials. The observed difference in pro- and anti-saccade latency and the presence of pro-saccade errors in the anti-saccade task indicates that a somatosensory stimulus can elicit a form of reflexive saccade comparable to pro saccades made in the visual modality. It is proposed that a peripheral somatosensory stimulus can elicit a form of reflexive saccade and that somatosensory saccades do not depend exclusively on higher level endogenous control processes for their generation. However, a comparison of the underlying latency distributions and of peak-velocity profiles of saccades made to visual and somatosensory stimuli showed that this distinction may be less clearly defined for the somatosensory modality and that modality-specific differences (such as differences in neural conduction rates) in the underlying oculomotor structures involved in saccade target selection also need to be considered. It is further suggested that a broader conceptualisation of saccades and saccade programming beyond the simple voluntary and reflexive dichotomy, that takes into account the control processes involved in saccade generation for both modalities, may be required. PMID- 16273410 TI - Intrauterine hemodynamics of tricuspid valve dysplasia in a growth-restricted infant. AB - We present a case of fetal tricuspid valve dysplasia, which was diagnosed at 22 weeks of gestation during a routine obstetrical examination. Serial fetal echocardiographic evaluation revealed progressive right ventricular outflow tract obstruction and persistent cardiomegaly. A female infant weighing 1,916 g was delivered by elective cesarean section at 38 weeks of gestation. Longstanding compression of the fetal lungs secondary to the persistent cardiomegaly resulted in severe respiratory distress and cyanosis immediately after birth. Although, mechanical ventilation and continuous infusion of prostaglandin were instituted, the infant died of respiratory failure at 21 days of age. During the pregnancy, the fetus exhibited intrauterine growth restriction, but hydrops did not occur. In regard to the fetal hemodynamics in this cardiac anomaly, transatrial communication is essential for fetal survival. The diameter of the fossa ovalis, which is a marker of transtrial blood flow, was adequate in this case. However, marked enlargement of the right heart associated with regurgitation interfered with left ventricular filling and output, which resulted in restriction of the combined ventricular output and intrauterine fetal growth restriction. PMID- 16273409 TI - Reproduction elevates the corticosterone stress response in common fruit bats. AB - Changes in reproductive state or the environment may affect the sensitivity of the hypothalamic-pituitary-andrenal (HPA) axis. However, little is known about the dynamics of the resulting corticosteroid stress response, in particular in tropical mammals. In this study, we address the modulation of corticosterone release in response to different reproductive conditions and seasonality in 326 free-living common fruit-eating bats (Artibeus jamaicensis) on Barro Colorado Island in Panama during dry and wet seasons. We present strong evidence that stress sensitivity is primarily modulated by reproductive condition. In reproductively active females, corticosterone increases were more rapid and reached higher levels, but also decreased significantly faster than in inactive females. The corticosterone response was weaker in reproducing males than in females and delayed compared to non-reproductive males. Testes volume in reproductively active males was negatively correlated with corticosterone concentrations. Our findings suggest differentiated dynamics in the corticosterone stress response between sexes, potentially reflecting conflicting ecological demands. In females, a strong acute corticosterone response may represent high stress- and risk-sensitivity that facilitates escape and thus helps to protect reproduction. In males, suppression during reproductive activity could reflect lowered stress sensitivity to avoid chronically elevated corticosterone levels in times of frequent aggressive and therefore costly inter male encounters. PMID- 16273406 TI - Influence of motor unit properties on the size of the simulated evoked surface EMG potential. AB - The purpose of the study was to quantify the influence of selected motor unit properties on the simulated amplitude and area of evoked muscle potentials detected at the skin surface. The study was restricted to a motor unit population simulating a hand muscle whose potentials were recorded on the skin over the muscle. Peak-to-peak amplitude and area of the evoked potential were calculated from the summed motor unit potentials and compared across conditions that simulated variation in different motor unit properties. The simulations involved varying the number of activated motor units, muscle fiber conduction velocities, axonal conduction velocities, neuronal activation times, the shape of the intracellular action potential, and recording configurations commonly used over hand muscles. The results obtained for the default condition simulated in this study indicated that ~7% of the motor unit potentials were responsible for 50% of the size of the evoked potential. Variation in the amplitude and area of the evoked muscle potential was directly related to the number of active motor units only when the stimulus activated motor units randomly, and not when activation was based on a parameter such as motor unit size. Independent adjustments in motor unit properties had variable effects on the size of the evoked muscle potential, including when the stimulus activated only a subpopulation of motor units. These results provide reference information that can be used to assist in the interpretation of experimentally observed changes in the size of evoked muscle potentials. PMID- 16273411 TI - Early pregnancy loss and neonatal deaths associated with Klebsiella pneumonia infection: a mini review of possible occupational health risk. AB - Recurrent pregnancy loss is a disease of grave psychological and economic concern. The etiology in the vast majority of the cases is unknown or at best poorly understood. Although Klebsiella pneumonia infections have been reported in humans and animals during pregnancy, there is hardly any information to indicate whether or not these infections may be responsible for early pregnancy loss. We present a review of literature and report for the first time in humans, Klebsiella pneumonia infection in placenta of a 38-year-old secondary recurrent aborter (parity 2 + 3). PMID- 16273412 TI - Renal hypoplasia: lessons from Pax2. AB - The functions of Pax2 during renal development are many. It organizes caudal descent of the nephric duct, emergence of the ureteric bud, branching morphogenesis, and sustained arborization of the collecting system. In this review, we use lessons from the study of Pax2 as organizing principles to focus on the developmental processes which, if disrupted, might lead to renal hypoplasia in humans. We consider the problem of renal hypoplasia as a continuum, ranging from renal agenesis to subtle congenital nephron deficits. Early failure in the first two developmental stages (e.g. homozygous inactivation of Pax2) should preclude formation of metanephric kidneys and cause bilateral renal agenesis, incompatible with life. Interference with the later stages affects the extent of branching morphogenesis (e.g. heterozygous Pax2 mutations). Although the resulting nephron deficits are compatible with life, they may be moderately severe and account for up to 40% of the children in dialysis and transplant units around the world. Finally, the effect of Pax2 on apoptosis in the branching ureteric bud seems to imply a quantitative process which is finely tuned. Modest changes in this program could account for subtle nephron deficits in "normal" humans and increased risk of hypertension or susceptibility to acquired renal disease later in life. PMID- 16273414 TI - Total persistence of thyroglossal duct with direct communication between cyst and foramen caecum. AB - During embryonic development the thyroid gland migrates through the thyroglossal duct from the pharyngeal endoderm to the anterior cervical region. The final step in this process is the total obliteration of the thyroglossal duct. A case is presented of a patient with a thyroglossal cyst together with a complete failure of the obliteration of the duct, which caused regurgitations of mucopurulent material after the expression of the cyst. This indicates a complete failure of the obliterative process. To the best of our knowledge, this phenomenon has not been described previously in the literature. PMID- 16273413 TI - Ultrasound of the abdomen and total bone scintigraphy in patients with cancer of the head and neck. AB - In this prospective study, the patients with head and neck cancer admitted to the Department of Otolaryngology and Head and Neck Surgery, Haukeland University Hospital, underwent examination with ultrasound of the abdomen (112 patients) and total bone scintigraphy (118 patients) in the primary staging of the disease and before the start of treatment. In only one patient did we find metastases of head and neck cancer in the liver by these additional investigations, and one patient was found to have bone metastases. No second primary cancers were found by these extra examinations. One patient was found to have an asymptomatic aorta aneurysm and was in need of urgent surgical treatment. Based on these findings, we nowadays only use ultrasound of the abdomen and total bone scintigraphy as part of the general examination of new head and neck patients in selected cases. PMID- 16273415 TI - Conservative management of significant epidural haematomas in children. AB - OBJECTIVE: A significant epidural haematoma (EDH) is generally treated by craniotomy and evacuation. This is a report of conservative management following an EDH on computerized tomography (CT) in a paediatric population. The authors examined whether conservative treatment of radiologically significant EDH is a successful and safe therapeutic option. METHODS: Retrospective data were collected from charts of patients with conservatively treated EDH in the Department of Surgery of the University Children's Hospital Zurich between September 1993 and January 2004. Included were patients without focal neurological deficits, with a Glasgow Coma Scale (GCS) of 15 and an initial CT demonstrating an EDH with a minimal thickness of 1 cm. Mild clinical symptoms of raised intracranial pressure such as headache, nausea or vomiting were treated symptomatically. Follow-up included a standardized interview, a neuropaediatric examination and CT. RESULTS: Thirteen children with EDH had successful conservative management. Only one 12-year-old female patient with a delayed diagnosed frontal EDH required surgical intervention 24 h after admission and 5 days after the accident. Clinical follow-up showed patients without neurological deficits, a Glasgow Outcome Scale of 5 and no post-traumatic sequelae over an average of 4 4/12 years (range 4 months to 10 4/12 years). Follow-up CT showed complete resolution of the EDH within 2 to 3 months. CONCLUSIONS: Our results demonstrate that significant EDH can be treated non-operatively in neurologically normal children. We recommend that such treatment be performed in specialised paediatric centres under adequate neurological observation since prompt emergency operation in case of neurological deterioration should be provided. PMID- 16273416 TI - Influence of a high-oxalate diet on intestinal oxalate absorption. AB - Hyperoxaluria is a major risk factor for renal stones. In most cases, it is sustained by increased dietary loads. In healthy individuals with a normal Western diet, the majority of urinary oxalate is usually derived from endogenous metabolism. However, up to 50% may be derived from the diet. We were interested in the effect of a high-oxalate diet on oxalate absorption, not merely on the frequently studied increased oxalate excretion. In study I, 25 healthy volunteers were tested with the [13C2]oxalate absorption test once while following a low oxalate (63 mg) and once while following a high-oxalate (600 mg) diet for 2 days each. In study II, four volunteers repeated study I, and afterwards continued with a high-oxalate diet (600 mg oxalate/day) for 6 weeks. In the last week, the [13C2]oxalate absorption test was repeated. After 4 weeks of individual normal diet, the oxalate absorption test with a high-oxalate diet was performed again. The results of study I show that the mean [13C2]oxalate absorption under low oxalate diet was 7.9 +/- 4.0%. In the presence of oxalate-rich food, the percent absorption for the soluble labelled oxalate almost doubled (13.7 +/- 6.3%). The results of study II show that the mean [13C2]oxalate absorption of the four volunteers under low-oxalate diet was 7.3 +/- 1.4%. The absorption increased to 14.7+/-5.2% under 600 mg oxalate. After 6 weeks under a high-oxalate diet, the [13C2]oxalate absorption was significantly decreased (8.2 +/- 1.7%). After the wash-out phase, the absorption was again high (14.1 +/- 2.2%) under the 600 mg oxalate challenge. PMID- 16273418 TI - Treating erectile dysfunction by endothelial rehabilitation with phosphodiesterase 5 inhibitors. AB - A large body of evidence has accumulated demonstrating that a common pathway in conditions such as hypertension, atherosclerosis, hypercholesterolemia, diabetes mellitus, and erectile dysfunction (ED) is endothelial dysfunction. Although a complete pharmacological cure for ED is currently unavailable, the phosphodiesterase 5 (PDE5) inhibitors sildenafil, vardenafil, and tadalafil are efficacious oral therapy for ED. Results from recent studies suggest that regular treatment with a PDE5 inhibitor may lead to enhanced erectile function (EF) beyond that observed with on-demand usage, possibly through improvement of endothelial function. Such an effect may be viewed as rehabilitation of damaged erectile tissue. The present review focuses on several recent studies which provide evidence for the beneficial effect of regular PDE5 inhibitor administration on the improvement of EF by rehabilitation of vascular endothelium. PMID- 16273417 TI - Clinical update on phosphodiesterase type-5 inhibitors for erectile dysfunction. AB - Erectile dysfunction (ED) affects the sexual lives of millions of men. The first line oral pharmacotherapy for most ED patients is phosphodiesterase type-5 (PDE 5) inhibitors, of which three are available. Sildenafil is the most widely prescribed oral agent for ED and has a very satisfactory efficacy-safety profile in all patient categories. Tadalafil and vardenafil were introduced in the European Union and in the United States in 2003 and 2004, respectively. The three PDE-5 inhibitors share many pharmacological and clinical characteristics, and each has unique features. This review, which is based on the contemporary literature on PDE-5 inhibitors, describes the chemical, pharmacological, and clinical features of sildenafil, vardenafil, and tadalafil. The first section reviews the pathophysiology of penile erection and PDE-5 inhibitor pharmacology. The second section summarizes data regarding efficacy and safety of the three drugs in treating ED in the general population as well as in selected patient categories. PMID- 16273419 TI - In vitro functional responses of isolated human vaginal tissue to selective phosphodiesterase inhibitors. AB - Only little is known as to the significance of the cyclic nucleotide-mediated signal transduction in the control of the function of human vaginal smooth musculature. Recently, the presence of the phosphodiesterase (PDE) isoenzymes 4 (cAMP-PDE) and 5 (cGMP-PDE) in the human vagina was reported. Thus, it was the aim of the study to elucidate the effects of some PDE inhibitors on the tension induced by endothelin 1 (ET-1), as well as on levels of cGMP and cAMP in isolated human vaginal wall tissue. Using the organ bath technique, the ability of norepinephrine (NE), carbachol, serotonin (5-HT), oxytocin and ET-1 to contract isolated vaginal wall muscle strips was evaluated. In another set-up, the effects of the PDE4 inhibitor rolipram and PDE5 inhibitors sildenafil and vardenafil (1 nM-10 microM) on the tension induced by 0.1 microM ET-1 of human vaginal wall tissue strips were investigated. In order to measure drug effects on tissue levels of cGMP and cAMP, vaginal tissue was exposed to different concentrations (0.1, 1 and 10 microM) of the compounds and the accumulation of cyclic nucleotides was determined. The adenylyl cyclase stimulating agents forskolin and nitric oxide donor sodium nitroprusside (SNP) (0.01, 0.1 and 1 microM) were used as reference compounds. While NE, carbachol and oxytocin failed to contract the vaginal tissue, ET-1 and, to a certain degree, 5-HT elicited contractile responses of the isolated strip preparations. The tension induced by 0.1 microM ET-1 was dose-dependently reversed by the drugs. The rank order of efficacy was sildenafil > forskolin > rolipram >or= vardenafil > SNP. Rmax values ranged from 24% (SNP) to 50% (sildenafil). With sildenafil being the only exception, none of the compounds reached an EC50 value. The relaxing effects of the drugs were paralleled by a fourfold to tenfold increase in tissue levels of cGMP and/or cAMP. Our results demonstrate that PDE inhibitors can relax human vaginal tissue and increase levels of cyclic nucleoside monophosphates. The findings with regard to the PDE5 inhibitors may indicate that the NO-cGMP pathway is, to a certain degree, involved in the control of vaginal smooth muscle tone. This might be of significance with regard to the pharmacological treatment of disorders connected with female sexual arousal and the ability to achieve orgasm. PMID- 16273420 TI - Dihydroartemisinin enhances radiosensitivity of human glioma cells in vitro. AB - PURPOSE: The antimalarial agent, artemisinin, also confers cancer-specific cytotoxic effects by reacting with ferrous iron atoms to form free radicals. Here, we investigated the radiosensitizing effects of dihydroartemisinin on glioma cells and assessed some possible mechanisms for these effects. MATERIALS AND METHODS: U373MG glioma cells treated with various concentrations of dihydroartemisinin plus radiation, and efficiency of radiosensitization was assessed by clonogenic survival assay. Expression and activity of antioxidant enzymes, glutathione-S-transferase (GST) were quantified by western blot and enzymatic activity analyses, respectively. RESULTS: Dihydroartemisinin showed higher cytotoxicity in the glioma cell lines than in the liver, breast or cervical cancer cell lines. In clonogenic survival assays, treatment with dihydroartemisinin alone dose-dependently reduced the number of U373MG colonies, while treatment with dihydroartemisinin plus gamma-irradiation showed far lower clonal survival than cultures treated with radiation or dihydroartemisinin alone. The radiosensitizing effect of dihydroartemisinin was blocked significantly by the free radical scavengers, NAC and TIRON, indicating association with dihydroartemisinin-induced ROS generation. In addition, the radiation-induced expression of endogenous GST was suppressed by treatment with dihydroartemisinin. The radiosensitizing effect of dihydroartemisinin was also markedly enhanced by the addition of holotransferrin CONCLUSION: Taken together, our results strongly suggest that dihydroartemisinin triggers production of ROS and inhibits GST activity, leading to effective and therapeutically relevant radiosensitization of human glioma cells. PMID- 16273421 TI - Effect of air leak on the performance of auto-PAP devices: a bench study. AB - Automatic positive airway pressure (auto-PAP) devices, used in the treatment of patients with obstructive sleep apnea (OSA), may not function optimally in the presence of an air leak. We set out to determine the magnitude of air leak that prevents auto-PAP devices from responding to respiratory events of OSA in a bench model. We simulated apnea, flow limitation, obstructive hypopnea, nonobstructive hypopnea, and snoring events of OSA with an artificial airway and a loudspeaker in a bench model connected to a commercially available auto-PAP device. Four auto PAP devices were tested, but two of the "older-generation" devices (Tranquility and Virtuoso) did not respond to events of OSA that involved changes in flow contour; hence, we studied the effects of air leak and humidifier in the two "newer-generation" auto-PAP devices only (GoodKnight 418P, Autoset-T). When the air leak was progressively increased from baseline levels recommended by the manufacturer to levels seen clinically--5 to 7, 10, and 30 l/min--the GoodKnight 418P decreased pressure response by 56% (5.6+/-1.8 cm H(2)O, p=0.04). The pressure response of the Autoset-T, however, did not change from baseline during similar levels of air leak. The GoodKnight performed appropriately when the air leak was within 20 l/min, but the corresponding value for the Autoset was higher at 50 l/min. For both devices combined, air leak caused the pressure to drop between the device and the airway: 2.8+/-0.3 cm H(2)O at 30 l/min of air leak (p<0.001). Air leaks cause auto-PAP devices to underestimate the pressure required to treat events of OSA and to overestimate the pressure delivered at the upper airway. Physicians should be aware of performance limitations of auto-PAP devices in the presence of an air leak. PMID- 16273422 TI - Activity levels of bats and katydids in relation to the lunar cycle. AB - Animals are exposed to many conflicting ecological pressures, and the effect of one may often obscure that of another. A likely example of this is the so-called "lunar phobia" or reduced activity of bats during full moon. The main reason for lunar phobia was thought to be that bats adjust their activity to avoid predators. However, bats can be prey, but many are carnivorous and therefore predators themselves. Thus, they are likely to be influenced by prey availability as well as predation risk. We investigated the activity patterns of the perch hunting Lophostoma silvicolum and one of its main types of prey, katydids, to assess the influence of the former during different phases of the lunar cycle on a gleaning insectivorous bat. To avoid sampling bias, we used sound recordings and two different capture methods for the katydids, as well as video monitoring and radio-telemetry for the bats. Both, bats and katydids were significantly more active during the dark periods associated with new moon compared to bright periods around the full moon. We conclude that foraging activity of L. silvicolum is probably influenced by prey availability to a large extent and argue that generally the causes of lunar phobia are species-specific. PMID- 16273424 TI - Studies of S-adenosylhomocysteine-hydrolase polymorphism in a Croatian population. AB - Recently, a proven case of human S-adenosylhomocysteine-hydrolase (SAHH) deficiency was reported in a Croatian boy. As molecular analysis of the SAHH gene in this case revealed two different mutant alleles, we investigated the polymorphism of human SAHH in a total of 237 red blood samples from unrelated Croats using starch gel electrophoresis and an enzyme-specific staining procedure. From the relative enzymatic activity of SAHH--determined by densitometric assessment of electrophoretic patterns, and calculated on the basis of the protein concentration of the red blood cells-we detected three individuals as being heterozygous for an SAHH 0-allele. Moreover, a total of four different electromorphic SAHHs have been observed, giving allele frequencies calculated as SAHH 1 = 0.941, SAHH 2 = 0.032, SAHH 3 = 0.006, SAHH 4 = 0.015, and SAHH 0 = 0.006. PMID- 16273423 TI - Activation tagging approach in a model legume, Lotus japonicus. AB - We constructed T-DNA insertional lines of a model legume, Lotus japonicus, using a multifunctional vector for gene and exon activation tagging. The vector had the CaMV 35S promoter together with two additional enhancer elements, the start codon, and splice donor and acceptor sites facing the left border, in anticipation of the activation of T-DNA flanking genes and forced expression of flanking exons. The improved transformation technique yielded more than 3,500 lines, including 45 dominant mutant candidates with abnormal phenotypes with respect to aerial parts, nodules, and roots. Among the 44 selected lines, one copy of T-DNA was inserted into the genome of 37 lines (84%). The T-DNA flanking regions of seven lines were isolated by thermal asymmetric interlaced (TAIL)-PCR or reverse transcription (RT)-PCR, and the corresponding genomic clones were analyzed. The transcripts of four genes adjacent to T-DNA out of 11 genes tested were increased in the T(1) generation, demonstrating that gene and exon activation effects by the newly developed tagging vector are heritable. The T-DNA insertional population of L. japonicus will provide legume-specific dominant mutants. PMID- 16273425 TI - California ground squirrel (Spermophilus beecheyi) defenses against rattlesnake venom digestive and hemostatic toxins. AB - Previous studies have shown that some mammals are able to neutralize venom from snake predators. California ground squirrels (Spermophilus beecheyi) show variation among populations in their ability to bind venom and minimize damage from northern Pacific rattlesnakes (Crotalus oreganus), but the venom toxins targeted by resistance have not been investigated. Four California ground squirrel populations, selected for differences in local density or type of rattlesnake predators, were assayed for their ability to neutralize digestive and hemostatic effects of venom from three rattlesnake species. In Douglas ground squirrels (S. b. douglasii), we found that animals from a location where snakes are common showed greater inhibition of venom metalloprotease and hemolytic activity than animals from a location where snakes are rare. Effects on general proteolysis were not different. Douglas ground squirrels also reduced the metalloprotease activity of venom from sympatric northern Pacific rattlesnakes (Crotalus oreganus oreganus) more than the activity of venom from allopatric western diamondback rattlesnakes (C. atrox), but enhanced fibrinolysis of sympatric venom almost 1.8 times above baseline levels. Two Beechey ground squirrel (S. b. beecheyi) populations had similar inhibition of venoms from northern and southern Pacific rattlesnakes (C. o. helleri), despite differences between the populations in the locally prevalent predator. However, the venom toxins inhibited by Beechey squirrels did vary among venom from Pacific rattlesnake subspecies, and between these venoms and venom from allopatric western diamondback rattlesnakes. Blood plasma from Beechey squirrels showed highest inhibition of metalloprotease activity of northern Pacific rattlesnake venom, general proteolytic activity and hemolysis of southern Pacific rattlesnake venom, and hemolysis by allopatric western diamondback venom. These results reveal previously cryptic variation in venom activity against resistant prey that suggests reciprocal adaptation at the molecular level. PMID- 16273426 TI - Chemical defense in harvestmen (arachnida, opiliones): do benzoquinone secretions deter invertebrate and vertebrate predators? AB - Two alkylated 1,4-benzoquinones were identified from the defensive secretion produced by the neotropical harvestman Goniosoma longipes (Gonyleptidae). They were characterized as 2,3-dimethyl-1,4-benzoquinone and 2-ethyl-3-methyl-1,4 benzoquinone. We tested the effectiveness of these benzoquinone secretions against several predator types, including invertebrates and vertebrates. Different predators were exposed to the harvestmen's gland secretion or to distilled water in laboratory bioassays. Our results indicate that secretions containing the 1,4-benzoquinones released by G. longipes can be an effective defense against predation, and that the effectiveness of the secretion is dependent on the predator type. The scent gland secretion repelled seven ant species, two species of large wandering spiders, and one frog species, but was not an effective defense against an opossum. Our study also demonstrates that the scent gland secretion of G. longipes can work as a chemical shield preventing the approach of three large predatory ants for at least 10 min. The chemical shield may protect the harvestman against successive attacks of the same ant worker and also allow the harvestman to flee before massive ant recruitment. Our data support the suggestion that chemical defenses may increase survival with some but not all potential predators. This variation in defense effectiveness may result from many interacting factors, including the attack strategy, size, learning ability, and physiology of the predators, as well as the chemical nature of the defensive compounds, type of emission, and amount of effluent released by the prey. PMID- 16273427 TI - Processing of a sesquiterpene lactone by Papilio glaucus caterpillars. AB - Papilio glaucus caterpillars encounter a diverse array of sesquiterpene lactones, including parthenolide, in the leaves of host plants Liriodendron tulipifera and Magnolia virginiana. These compounds are toxic to unadapted herbivores, and the development of P. glaucus caterpillars likely depends on their ability to excrete or detoxify them efficiently. A new metabolite of parthenolide, 2-alpha hydroxydihydroparthenolide, identified by crystal structure determination and nuclear magnetic resonance, was present in the waste of the caterpillars. The parent compound was modified by the reduction of an alpha-methylene group, rendering the compound less reactive, and the addition of a hydroxyl group, which increases the polarity and prepares it for the conjugation reactions of phase II metabolism. Unmetabolized parthenolide was also present in large amounts in waste. P. glaucus larvae are apparently capable of excreting intact sesquiterpene lactones and sesquiterpene lactone metabolites during consumption of foliage rich in these compounds. PMID- 16273428 TI - Similarity of cuticular lipids between a caterpillar and its host plant: a way to make prey undetectable for predatory ants? AB - Ithomiine butterflies (Nymphalidae) have long-lived, aposematic, chemically protected adults. However, little is known about the defense mechanisms in larvae and other juvenile stages. We showed that larvae Mechanitis polymnia are defended from ants by a chemical similarity between their cuticular lipids and those of the host plant, Solanum tabacifolium (Solanaceae). This is a novel defense mechanism in phytophagous insects. A field survey during one season showed that larval survivorship was up to 80%, which is high when compared with other juvenile stages. In a laboratory bioassay, live larvae on their host plant were not attacked by the predatory ant Camponotus crassus (Formicidae). Two experiments showed that the similarity between the cuticular lipids of M. polymnia and S. tabacifolium protected the larvae from C. crassus: (a) when the caterpillar was switched from a host plant to a non-host plant, the predation rate increased, and (b) when a palatable larva (Spodoptera frugiperda, Noctuidae) was coated with the cuticular lipids of M. polymnia and placed on S. tabacifolium leaves, it no longer experienced a high predation rate. This defensive mechanism can be defined as chemical camouflage, and may have a double adaptive advantage, namely, protection against predation and a reduction in the cost of sequestering toxic compounds from the host plant. PMID- 16273429 TI - Divergence in structure and activity of phenolic defenses in young leaves of two co-occurring Inga species. AB - The leaves of tropical forest trees are most likely to suffer herbivore damage during the period of expansion. Herbivore selection on young leaves has given rise to a variety of leaf developmental strategies and age-specific chemical defense modes. We are studying correlations between leaf developmental types and chemical defenses in the Neotropical genus Inga. We have characterized defense metabolites in Inga goldmanii and Inga umbellifera, two species that co-occur in the lowland moist forest of Panama. These congeners have markedly different young leaf developmental phenotypes but suffer approximately equal rates of herbivory. Bioassays of whole and fractionated leaf extracts using larvae of Heliothis virescens show that I. goldmanii chemical defenses are nearly three times more inhibitory than those of I. umbellifera. In both species, most of the inhibitory activity resides in complex mixtures of monomeric and polymeric flavan-3-ols. This group comprises >30% of young leaf dry weight in both I. goldmanii and I. umbellifera. The species' phenolic chemistry differs markedly, however, both in the structure of the monomeric units and in the distribution of polymer sizes. The differences in chemical structure have pronounced effects on their bioactivities, with I. goldmanii flavans being twice as inhibitory to H. virescens larvae as I. umbellifera flavans, and more than three times more efficient at protein binding. Given the extraordinarily high polyphenol concentrations that are found in the young leaves of these species, protein precipitation could be an important mechanism of growth inhibition. Nevertheless, our data show that another mode of phenolic action, possibly oxidative stress, occurs simultaneously. PMID- 16273430 TI - The effect of pollination on floral fragrance in thistles. AB - We investigated postpollination changes in fragrance composition and emission rates, as well as pollinator discrimination in hand-pollinated flower heads of two thistle species: Canada thistle (Cirsium arvense) and sandhill thistle (C. repandum). Following pollination, neither species emitted any novel compounds that could function as repellents. Scent emission rates declined in pollinated plants of both species by approximately 89% within 48 hr. This decline was evident in all 13 scent components of C. arvense. Apis mellifera, the dominant pollinator in the study population of C. arvense, was nearly three times more likely to visit an unpollinated rather than a pollinated flower head. A more complex pattern was observed for C. repandum, whose scent comprised 42 compounds. Quantities of aromatic and sesquiterpenoid volatiles declined after pollination, whereas two classes of scent compounds, fatty acid derivatives and monoterperpenoids, continued to be emitted. In C. repandum, discrimination against pollinated flower heads by Papilio palamedes (its primary pollinator) was not as marked. Unpollinated control plants of both species maintained moderate levels of scent production throughout this experiment, demonstrating that senescence and floral advertisement may be delayed until pollination has occurred. We expect postpollination changes in floral scent contribute to communication between plants with generalized pollinator spectra and their floral visitors. This study provides the first field study of such a phenomenon outside of orchids. PMID- 16273431 TI - Effects of drought stress and nutrient availability on dry matter allocation, phenolic glycosides, and rapid induced resistance of poplar to two lymantriid defoliators. AB - The growth-differentiation balance hypothesis (GDBH) postulates that variation in resource availability can increase or decrease allocation to secondary metabolism, depending on how growth is affected relative to carbon assimilation. Growth and leaf area of black poplar (Populus nigra) increased substantially in response to increased nutrient availability, while net assimilation rate and photosynthesis were less strongly affected. In response, total phenolic glycoside concentrations declined, which is consistent with GDBH. Drought stress decreased net assimilation rate and photosynthesis as well as growth, while increasing total phenolic glycoside concentrations. This pattern does not follow GDBH, which predicts lower secondary metabolism when resource limitation decreases both growth and carbon assimilation. However, there was a strong negative correlation between growth and total phenolic glycoside concentration consistent with a trade off between primary and secondary metabolism, a key premise of GDBH. Drought decreased the growth of gypsy moth (Lymantria dispar) larvae but had no effect on whitemarked tussock moth (Orgyia leucostigma). Increased nutrient availability had a positive linear effect on growth of whitemarked tussock moth, but no effect on gypsy moth. Treatment effects on gypsy moth corresponded closely with effects on total phenolic glycosides, whereas effects on whitemarked tussock moth more closely tracked changes in nutritional quality. Localized gypsy moth herbivory elicited rapid induced resistance to gypsy moth, with the effect being independent of water and nutrient availability, but did not affect whitemarked tussock moth, indicating that the effects of biotic and abiotic stress on insect resistance of trees can be species-specific. PMID- 16273433 TI - Changes in heartwood chemistry of dead yellow-cedar trees that remain standing for 80 years or more in southeast Alaska. AB - We measured the concentrations of extractable bioactive compounds in heartwood of live yellow-cedar (Chamaecyparis nootkatensis) trees and five classes of standing snags (1-5, averaging 4, 14, 26, 51, and 81 years-since-death, respectively) to determine how the concentrations changed in the slowly deteriorating snags. Three individuals from each of these six condition classes were sampled at four sites spanning a 260-km distance across southeast Alaska, and the influence of geographic location on heartwood chemistry was evaluated. Cores of heartwood were collected at breast height and cut into consecutive 5-cm segments starting at the pith. Each segment was extracted with ethyl acetate and analyzed by gas chromatography. Concentrations of carvacrol, nootkatene, nootkatol, nootkatone, nootkatin, and total extractives (a sum of 16 compounds) for the inner (0-5 cm from pith), middle (5-10 cm from pith), and surface (outer 1.1-6.0 cm of heartwood) segments from each core were compared within each tree condition class and within segments across condition classes. Heartwood of class 1 and 2 snags had the same chemical composition as live trees. The first concentration changes begin to appear in class 3 snags, which coincides with greater heartwood exposure to the external environment as decaying sapwood sloughs away, after losing the protective outer bark. Within core segments, the concentrations of all compounds, except nootkatene, decrease between snag classes 2 and 5, resulting in the heartwood of class 5 snags having the lowest quantities of bioactive compounds, although not different from the amounts in class 4 snags. This decline in chemical defense is consistent with heartwood of class 5 snags being less decay resistant than heartwood of live trees, as observed by others. The unique heartwood chemistry of yellow cedar and the slow way it is altered after death allow dead trees to remain standing for up to a century with a profound impact on the ecology of forests in southeast Alaska where these trees are in decline. PMID- 16273432 TI - Plant surface properties in chemical ecology. AB - The surface of the primary aerial parts of terrestrial plants is covered by a cuticle, which has crucial autecological functions, but also serves as an important interface in trophic interactions. The chemical and physical properties of this layer contribute to these functions. The cuticle is composed of the cuticular layer and the cuticle proper, which is covered by epicuticular waxes. Whereas the cutin fraction is a polyester-type biopolymer composed of hydroxyl and hydroxyepoxy fatty acids, the cuticular waxes are a complex mixture of long chain aliphatic and cyclic compounds. These highly lipophilic compounds determine the hydrophobic quality of the plant surface and, together with the microstructure of the waxes, vary in a species-specific manner. The physicochemical characteristics contribute to certain optical features, limit transpiration, and influence adhesion of particles and organisms. In chemical ecology, where interactions between organisms and the underlying (allelo-) chemical principles are studied, it is important to determine what is present at this interface between the plant and the environment. Several useful equations can allow estimation of the dissolution of a given organic molecule in the cuticle and its transport properties. The implementation of these equations is exemplified by examining glucosinolates, which play an important role in interactions of plants with other organisms. An accurate characterization of physicochemical properties of the plant surface is needed to understand its ecological significance. Here, we summarize current knowledge about the physical and chemical properties of plant cuticles and their role in interactions with microorganisms, phytophagous insects, and their antagonists. PMID- 16273434 TI - Impact of phenolic compounds and related enzymes in sorghum varieties for resistance and susceptibility to biotic and abiotic stresses. AB - Contents of phenolic compounds and related enzymes before and after sorghum grain germination were compared between varieties either resistant or susceptible to biotic (sooty stripe, sorghum midge, leaf anthracnose, striga, and grain molds) and abiotic (lodging, drought resistance, and photoperiod sensitivity) stresses. Independent of grain germination, sorghum varieties resistant to biotic and abiotic stresses had on average higher contents of proanthocyanidins (PAs), 3 deoxyanthocyanidins (3-DAs), and flavan-4-ols than susceptible varieties. Results show that content of 3-DAs is a good marker for sorghum resistance to both biotic and abiotic stresses because it correlates with resistance to all stresses except for photoperiod sensitivity. The second good marker for stress resistance is content of PAs. Total phenolic compounds and the activities of related enzymes are not good markers for stress resistance in sorghum grains. PMID- 16273435 TI - Whole plant response of lettuce after root exposure to BOA (2(3H) benzoxazolinone). AB - The goal of our work was to expand the knowledge about plant stress response to the allelochemical 2(3H)-benzoxazolinone (BOA). We focused on physiological processes that are affected by this secondary metabolite. Physiological and biochemical characteristics of plants exposed to BOA help us to better understand its mode of action and open the gate to the use of allelochemicals as "natural" herbicides. Measurements on photosynthesis, fluorescence, water relations, antioxidant enzymes (superoxide dismutase, peroxidase), ATPases, and lipid peroxidation indicated that a phytotoxic effect follows BOA exposition. This effect was intense enough to interfere with plant growth and development and to produce "induced senescence." Based on this, we propose a multifaceted mode of action for BOA with effects at different levels and in different parts of the plant. PMID- 16273436 TI - Pheromonal activity of compounds identified from male Phyllotreta cruciferae: field tests of racemic mixtures, pure enantiomers, and combinations with allyl isothiocyanate. AB - Four himachalene sesquiterpenes and (+)-gamma-cadinene, previously identified as possible pheromone components from males of a North American population of Phyllotreta cruciferae Goeze (Coleoptera, Chrysomelidae), were tested for attractiveness in field trapping experiments in Hungary. A mixture of the four synthetic racemic himachalene derivatives and (+)-gamma-cadinene from a botanical source was slightly attractive to beetles, but much more attractive when blended with the known host-plant-derived attractant allyl isothiocyanate. This result was consistent with a previous study in North America. In tests with optically pure synthetic compounds, a blend of the same himachalene enantiomers found from male beetles was equivalent to the corresponding blend of racemic compounds, whereas a blend of the opposite enantiomers was not active. Through subtraction tests, it was found that the single compound, (6R,7S)-2,2,6,10 tetramethylbicyclo[5.4.0.]undeca-9,11-diene [compound (+)-A in this study], was as active as the whole mixture, suggesting that this compound is the key pheromone component of the European population of P. cruciferae. During field trials, several congeneric species, including P. vittula, P. nemorum, P. nodicornis, and P. ochripes, also were caught, suggesting that the same compound(s) may be relatively widespread as pheromone components in this genus. PMID- 16273437 TI - Identification of a sex pheromone from male yellow mealworm beetles, Tenebrio molitor. AB - The sex pheromone released by the adult female Tenebrio molitor, 4-methyl-1 nonanol, is well known. In addition, there is evidence that adult males release a pheromone that attracts females. The purpose of the present study was to isolate and identify male-released pheromone(s). Emissions from virgin adult males and females were collected on filter paper and extracted with pentane. Extracts were analyzed by gas chromatography-mass spectrometry. One male-specific compound was detected and identified as (Z)-3-dodecenyl acetate (Z3-12:Ac). In arena bioassays, E3-12:Ac was attractive to females only, at 1 and 10 microg doses. E3 12:Ac was also attractive to females at a 10-microg dose. The presence of both male and female pheromones, each attracting the opposite sex, may contribute to maintaining a high-density population of both sexes. PMID- 16273438 TI - Pheromone communication in the honeybee (Apis mellifera L.). AB - Recent studies have demonstrated a remarkable and unexpected complexity in social insect pheromone communication, particularly for honeybees (Apis mellifera L.). The intricate interactions characteristic of social insects demand a complex language, based on specialized chemical signals that provide a syntax that is deeper in complexity and richer in nuance than previously imagined. Here, we discuss this rapidly evolving field for honeybees, the only social insect for which any primer pheromones have been identified. Novel research has demonstrated the importance of complexity, synergy, context, and dose, mediated through spatial and temporal pheromone distribution, and has revealed an unprecedented wealth of identified semiochemicals and functions. These new results demand fresh terminology, and we propose adding "colony pheromone" and "passenger pheromone" to the current terms sociochemical, releaser, and primer pheromone to better encompass our growing understanding of chemical communication in social insects. PMID- 16273439 TI - Becoming a research-oriented family physician. PMID- 16273440 TI - Ethics and spirituality are not synonyms. PMID- 16273441 TI - Continuity of care. PMID- 16273442 TI - Do resident work-hour regulations really increase patient safety? A resident's perspective. PMID- 16273443 TI - A new model of practice: implications for medical student teaching in family medicine. PMID- 16273444 TI - Allopathic family medicine residents can learn osteopathic manipulation techniques in a 1-month elective. AB - BACKGROUND: Graduating family medicine residents report a relative lack of confidence in managing musculoskeletal problems, and many primary care physicians desire more instruction in manual medicine. METHODS: We conducted a 1-month osteopathic manipulative treatment elective with five allopathic family medicine residents, utilizing multiple teaching and assessment strategies. RESULTS: Residents averaged 30 patient encounters each. Faculty graded their attainment of the knowledge and skills objectives at 3.9 and 3.8 on a 5-point scale, respectively. Residents reported unanimously that the course had reasonable expectations and fostered independent decision making and that they achieved the educational goals. CONCLUSIONS: After a 1-month elective, allopathic residents demonstrated competency in a defined set of osteopathic principles and skills. PMID- 16273447 TI - Poetry in the borderlands of medicine. PMID- 16273448 TI - Student and resident education and rural practice in the Southwest Indian Health Service: a physician survey. AB - BACKGROUND AND OBJECTIVES: The Indian Health Service (IHS) is an educational rotation site for numerous medical students and residents. These IHS rotations may be an important factor in recruitment and retention of physicians to the IHS. We describe the combined number of student/resident rotations in the Southwest IHS and their influence on recruitment and retention. We also analyze factors related to choice of rural practice in the IHS. METHODS: We conducted a survey of clinical directors and IHS physicians in Arizona and New Mexico. RESULTS: Twenty (87%) clinical director surveys and 289 (66%) physician surveys were returned. More than 400 students/residents participate in rotations annually in the IHS in Arizona and New Mexico. Eighty-four percent of clinical directors feel that educational programs are important to recruitment. Forty-five percent of current IHS physicians participated in IHS rotations as students or residents, and 87% feel that rotating influenced their decision to join the IHS. Eighty percent of IHS physicians who teach feel that working with students and residents improves their job satisfaction. Seventy-five percent of respondents practice in rural areas. Rural medical student and resident rotations are associated with subsequent rural practice. CONCLUSIONS: Many medical students and residents rotate in the Southwest IHS. Clinical directors state that these rotations are helpful to recruitment, and IHS physicians who rotated feel it was important in their decision to join the IHS. IHS clinicians feel that teaching improves job satisfaction. PMID- 16273449 TI - The current state of flexible sigmoidoscopy training in family medicine residency programs. AB - BACKGROUND AND OBJECTIVES: The US Preventive Services Task Force has recommended that adults ages 50 and over be screened for colorectal cancer. Flexible sigmoidoscopy (FS) is one available screening option. This study determined the current state of FS training in US family medicine residencies. METHODS: Directors of the Accreditation Council for Graduate Medical Education-accredited family medicine residencies were surveyed regarding FS training. RESULTS: Of 486 mailed surveys, 370 (76%) were completed and returned. Fifty-two percent of responding residency programs trained at least one resident in FS in 2003. Residents in these programs performed a mean of 20.1 +/- 1.2 FSs during their training. In 2003, 44% of family medicine graduates from these programs were certified by their programs as competent to perform FS. Fewer residents were certified in FS by programs in the eastern versus western United States. Military programs certified more residents than did nonmilitary programs. CONCLUSIONS: More than half of programs offered FS training, but less than half of family medicine graduates were certified by their programs as competent. There were significant differences for FS training by region and program type. PMID- 16273451 TI - Are patients who use a generalist physician healthier than those who seek specialty care directly? AB - BACKGROUND: American health care consumers want the option of seeing specialists whenever they wish, but given this option, do they in fact use it without consideration of their health status? This paper reports on a cross-sectional analysis that compares the demographics and health status of fee-for-service Medicare enrollees who exhibited four different patterns of physician access. METHODS: The Medicare Beneficiary Survey data from 1998 were used. Subjects ages 65 and older were categorized into one of four groups: those with no physician claim, those who saw a generalist only, those who saw a specialist only, and those who saw both. Age, income, education, health status, level of impairment, and disease burden for the four patient groups were compared using ANOVA. Urban/rural status, race, ethnicity, mortality rates, and gender for the four patient groups were compared using chi-square. A predictive model using mutinomial logistic regression was created. RESULTS: Twelve percent of subjects saw no physician in 1998, 11.6% saw a generalist only, 14.2% visited a specialist only, and 62.1% visited both types of physicians. Subjects who saw both physician types had significantly worse health status and more chronic diseases than the other groups. Subjects who saw generalists only or specialists only had intermediate levels of health status and disease burden that were not significantly different from each other. Subjects who saw a specialist only were the most affluent and highly educated group. Subjects who saw no physician had the best health status and the fewest chronic diseases of all subject groups. Urban residents were more likely to visit some type of physician than were rural residents and were more likely to see a specialist only. Regional differences were noted, with New England showing the highest rates of specialist only use. DISCUSSION: As expected, the healthiest subjects were least likely to visit any health care provider. Subjects with the worst health status were likely to access both generalists and specialists for their care. Subjects who visited a specialist only had higher incomes, more education, and urban residence but no difference in health status when compared to subjects who visited a generalist only. PMID- 16273450 TI - Correlates of medication knowledge and adherence: findings from the residency research network of South Texas. AB - BACKGROUND: Medication adherence is a complex phenomenon, influenced by a variety of factors. Most adherence research focuses on one medicine and does not represent the realities of clinical family medicine. This analysis examined factors associated with medication knowledge and adherence in family medicine patients with chronic conditions. METHODS: The Residency Research Network of South Texas (RRNeST) enrolled 150 patients with chronic disease who "sometimes have trouble taking medicines." Seventy-five percent were Latinos. This cross sectional analysis used baseline survey data from an intervention study. Investigators correlated medication knowledge and adherence with known predictors -patient, health, medication, economic, and physician factors. New variables related to patients' motivation to change treatment behaviors ("importance" and "confidence") were also included. RESULTS: Linear regression analysis demonstrated that patient satisfaction, education level, and confidence were associated with better medication knowledge. Higher confidence, Spanish language, better functional and health status, and more prescription medicines were correlated with medication adherence. CONCLUSIONS: We recommend that family physicians enhance medication adherence by providing good information about treatment and counseling strategies to build patients' confidence. Our findings suggest that poor health status can be a barrier to, rather than a motivator for, treatment adherence. PMID- 16273452 TI - Are personal digital assistants an acceptable incentive for rural community-based preceptors? AB - BACKGROUND AND OBJECTIVES: This study's objective was to evaluate the acceptability, effect, and use of handheld computers (also known as personal digital assistants or PDAs) as a reward for undergraduate rural community-based family medicine preceptors. METHODS: All rural, undergraduate family physician teachers who accepted an undergraduate student for a 1-month placement were offered the choice between a PDA that carried medical software or a monetary payment of an equivalent value. Approximately 1 year later, different surveys were sent to both groups of preceptors to collect data on their use of PDAs and computer technology. RESULTS: The most commonly reported reason for choosing a PDA in lieu of payment was that it provided a good opportunity to learn about PDA technology. Of those who accepted a PDA, however, 10% had not yet used it, and another 44% of recipients had difficulty in getting started using the PDA. There were more reported problems with the software than the hardware. When surveyed 1 year later, those who received a PDA and were still using it reported satisfaction with the medical software, ranging from 31% for Epocrates qid to 71% for the 5-Minute Medical Consult. More than 90% of those using their PDA 1 year later reported that they used it in clinical settings, with 68% feeling their PDA had some or a significant effect on patient care. CONCLUSIONS: Rural family physicians appeared to find PDAs an acceptable reward for teaching, based on the reported use and utility of their PDA, but many had technical difficulties. Recipients of the PDA reported using their PDA primarily in the clinical setting, with the feeling that the PDA had a positive effect on their patient care. Many users had difficulty with technical aspects of PDA use. To support PDA recipients, technical assistance should be provided. PMID- 16273453 TI - Effect of a PDA-assisted evidence-based medicine course on knowledge of common clinical problems. AB - BACKGROUND AND OBJECTIVES: It is not yet known if personal digital assistant (PDA)-assisted evidence-based medicine (EBM) courses in postgraduate training enhance knowledge of common clinical problems. This study's objective was to determine if PDA-assisted EBM training would improve clinical knowledge. METHODS: In a controlled trial, intervention group residents received InfoRetriever on a PDA coupled with an EBM course integrated within clinical rotations in family medicine. The effect of the intervention and the rate of use of InfoRetriever on a written test of knowledge were evaluated after adjusting for baseline knowledge scores. The test measured knowledge of primary care management of hypertension and diabetes as well as estimation of disease probability. RESULTS: There was no effect on first posttest knowledge scores of the intervention overall or of the rate with which participants had used InfoRetriever during the intervention. However, when intervention group residents retook the test with access to InfoRetriever while taking the knowledge test, scores increased 7.4% (+2.4 correct test questions). Access to InfoRetriever Clinical Prediction Rules on a PDA, however, had an unclear effect on residents' ability to estimate disease probability. CONCLUSIONS: There was no effect of a PDA-assisted EBM course on knowledge test scores, although using the PDA during the test results in higher scores. It is unclear if using PDA Clinical Prediction Rules can improve residents' estimates of disease probability. PMID- 16273454 TI - Will technology make us better educators and clinicians? PMID- 16273455 TI - Sigmoidoscopy versus colonoscopy: ask yourself. PMID- 16273458 TI - [Electrophysiological estimation of the peripheral nerves conduction parameters and the autonomic nervous system function in the course of amyotrophic lateral sclerosis]. AB - BACKGROUND AND PURPOSE: The aim of the study was to electrophysiologically examine the peripheral nervous system, mainly its autonomic part in the course of amyotrophic lateral sclerosis (ALS). MATERIAL AND METHODS: 19 patients with clinically definite or probable ALS were examined twice (group A and B). 20 healthy subjects were included in the study (group C). Motor conduction study (ulnar, peroneal nerve) with F wave examination, sensory conduction study (ulnar, sural nerve), EMG from 4 muscles, sympathetic skin response (SSR) and heart rate variability test at rest were performed. RESULTS: In the motor conduction study we revealed a significant lowering of the muscle action potential amplitude in the patient group compared with controls and its lowering in the course of ALS. F wave latencies were significantly longer in patients when compared with controls with no progression in the course of ALS. Sensory conduction parameters and heart rate variability parameters at rest did not differ significantly in groups A, B and C. SSR amplitudes were significantly lower and SSR latencies were significantly longer in the patient group in comparison with the control group. There was a significant progression of these parameters during the ALS course. CONCLUSIONS: Loss of motor fibres creating ventral roots and sudomotor fibres lesion are the integral parts of the ALS clinical picture. They tend to worsen during the ALS course. PMID- 16273459 TI - [Analyses of 14-3-3 protein in the cerebrospinal fluid in Creutzfeldt-Jakob disease. Preliminary report]. AB - BACKGROUND AND PURPOSE: Definitive diagnosis of Creutzfeldt-Jakob disease (CJD) is possible only after neuropathological confirmation; there are no non-invasive tests which allow to definitely diagnose CJD pre mortem. The only one biochemical marker included in the diagnostic criteria for CJD approved by WHO is an elevated level of 14-3-3 protein in the cerebrospinal fluid (CSF). The aim of the study was to introduce 14-3-3 analysis for a routine diagnostics of CJD in Poland and to optimise the Western blotting technique used in the Reference Centre of Prion Diseases in Lodz. MATERIAL AND METHODS: We analysed 33 samples of CSF: 17 from patients suspected of CJD (2 definitive, 2 probable, 4 possible, 11 others), and 16 controls (from patients with other neurological diseases). CSF samples were analysed using the Western blotting method and commercially available antibodies (Santa Cruz). We used 2 methods of CSF preparation: with and without lyophilization, and 2 different anti-14-3-3 antibodies: rabbit polyclonal and mouse monoclonal antibodies. RESULTS: We found 14-3-3 protein in both definitive sCJD cases and in one probable case. Among other samples, we obtained a positive result only in one patient with alcohol abuse history and in samples of CSF contaminated with erythrocytes. CONCLUSIONS: The Western blotting test for 14-3-3 protein in CSF is useful for diagnosis of CJD, but only when considered in an appropriate clinical context, together with other diagnostic criteria. Contamination of CSF with erythrocytes can cause false positive results. Analysis of a non-lyophilised CSF and detection of 14-3-3 protein using the mouse monoclonal antibody gave the most reliable results. PMID- 16273460 TI - ["Malignant" middle cerebral artery territory infarction]. AB - The pathology, clinical course, outcome, diagnosis, treatment and prognosis of dramatic malignant middle cerebral artery territory infarction were presented. About 10% of stroke patients suffer from malignant middle cerebral artery territory infarction, mainly due to brain edema and herniation. This syndrome causes high mortality. The newest conservative and surgical treatment was presented. PMID- 16273461 TI - [Attention impairments in aphasia. Psychological aspects]. AB - This paper reviews the relationship between the attention impairment and language functions in aphasia. It contains the description of some experiments with methods useful to examine the functions of attention: divided, selective, focused attention, suitable for patients with aphasia. The paper summarises the growing literature which documents the influence of divided attention on language functions such as: word retrieval, comprehension, spoken language, picture description. Lastly, the clinical implications of adopting the attentional model of aphasia are considered with respect to strategies for both assessment and treatment of adults with aphasia. PMID- 16273462 TI - [Sleep disturbances in Parkinson's disease]. AB - Parkinson's disease is a progressive disorder of the central nervous system. Degeneration of the dopaminergic neurons is the main cause of the disease. The basic symptoms of Parkinson's disease are bradykinesia, rigidity and resting tremor. Disturbances of the autonomous nervous system, depression, dementia and sleep disorders are common, too. People with Parkinson's disease suffer from insomnia, excessive daytime sleepiness, "sleep attacks", nightmares, REM sleep behaviour disorder, periodic limb movement in sleep, restless legs syndrome and sleep apnea syndrome. The main cause of sleep disorders in Parkinson's disease are age-connected changes in sleep architecture, disturbances of neurotransmission, movement disturbances in sleep, medications and concomitant diseases. The authors present the current state of knowledge on sleep disorders in Parkinson's disease, especially, the role of dopaminergic therapy, methods of diagnostics and treatment as well as the influence of sleep disturbances on patient's quality of life. PMID- 16273463 TI - [The diagnostic and therapeutic application of transcranial magnetic stimulation]. AB - The functional abnormalities of the central motor structures and its contribution of rigidity, tremor and bradykinesia in Parkinson's disease seem mainly due to the degeneration of the nigro-striatal pathway. Transcranial magnetic stimulation (TMS) of the motor cortex may provide useful data on the pathophysiology of motor dysfunction in Parkinson's disease. Recent reviews on the basic mechanisms of TMS in Parkinson's disease show reduced inhibitory motor network at the cortical and spinal level. The observed changes are thought to be in relation with a dysfunction of subcortico-cortical and subcortico-spinal pathways. The abnormalities of the central motor function seem to be modified by several clinical related factors as prevalence of cardinal Parkinson's disease signs (e.g. rigidity versus tremor or bradykinesia), L-dopa therapy ('on/off' states) and laterality of the Parkinson's disease signs. Observations made using TMS give new pathophysiological insights in functioning of the central motor structures in Parkinson's disease and started new form of TMS - repetitive TMS (rTMS) as a treatment of the Parkinson's disease motor signs. A few studies using rTMS with repetition rate of 0.2, 1, and 5 Hz showed improvement of motor signs in the Parkinson's disease patients. Although these results support the beneficial effects of rTMS on parkinsonian symptoms, long-term studies with large numbers of subjects should be conducted to assess the efficacy of the rTMS on Parkinson's disease in future. PMID- 16273465 TI - [A case of a giant cavernoma located in the temporal region]. AB - We present a case of a 21-year-old female with a giant cavernoma (5 cm x 3.1 cm x 3.6 cm) located in the left temporal region. Giant cavernomas are very rarely diagnosed. In our research we wanted to focus on the occurrence of these vascular malformations and diagnostic problems related to this matter. PMID- 16273464 TI - [The role of protein conformational disturbances in the pathomechanism of the extrapyramidal system diseases]. AB - Neurodegenerative disorders are characterised by cell damage due to accumulation of toxic, pathologic proteins. Mutations in genes coding different cell proteins result in conformational disturbances of the protein structure, their accumulation and aggregation manifested at the level of light microscope as various intracellular inclusions. This paper is an attempt of approach to cellular mechanisms underlying neurodegenerative diseases of the extrapyramidal system with special attention to ubiquitin-proteasome pathway -- the pathway whose discoverers received the 2004 Nobel Prize in chemistry. PMID- 16273466 TI - [A rare case of chondromyxoid fibroma of the parietal bone]. AB - The authors present a very rare case of the chondromyxoid fibroma (CMF) of the parietal bone. This is an uncommon chondroid tumour which constitutes less than 1% of primary bone tumours and which is usually localised in the metaphysis of the long bones, often in the knee region. There are 23 cases of cranial localisation of the chondromyxoid fibroma reported in several papers, 14 cases involved the cranial base and 9 the calvaria. A different ossification process of the skull-base and calvaria is probably responsible for this distribution. In our case we found the focal, lytic lesion in the parietal bone and fibro-greasy tumour mass. The dura was spared. The tumour was removed totally. Curettage of these kind of lesions should be avoided because it may lead to tumour recurrence. Histologically this tumour may cause problems in a differential diagnosis with other chondroid tumours like chondrosarcoma or chondroblastoma. PMID- 16273467 TI - [Guillain-Barre syndrome in a course of early cutaneous type of Lyme borreliosis: diagnostic and therapeutic difficulties]. AB - A case is reported of a 33-year-old man in whom Guillain-Barre syndrome (GBs) developed three weeks after a tick's sting. At the sting site typical for an early cutaneous type of Lyme borreliosis -- erythema migrans -- appeared. The demyelinating polyradiculoneuropathy of GBs occurred after disappearance of erythema migrans, and was manifested by progressive neuropathic symmetrical limb weakness with distal numbness and pain, and bilateral facial paralysis. The GBs was confirmed by electrophysiological examination and elevated protein concentration with a normal range of cells in the cerebrospinal fluid. Antibodies IgM and IgG against Borrelia burgdorferi in the blood serum and cerebrospinal fluid assessed using immunoenzymatic assay, MEIA, were negative on account of their early search. The above findings suggested that the GBs appearance after the probable Borrelia burgdorferi infection was in fact due to that infection. The patient recovered after treatment with plasma-phoresis and corticosteroid therapy followed by intravenous immunoglobulin, and physiotherapy. This is the first case in the Polish neurological literature of GBs with an early skin form of borreliosis which developed after the tick's sting. PMID- 16273468 TI - [Spinal cord stimulation in refractory angina pectoris -- a case description]. AB - Spinal cord stimulation (SCS) is one of the methods for treating angina pectoris and has been used since 1985. Patients with refractory angina pectoris despite optimal medication and revascularisation procedures are suitable for the SCS. This kind of treatment is not common in Poland. This article reports the first case of a patient with ischaemic heart disease treated with spinal cord stimulation in the Clinical Department of Neurosurgery in the Military Clinical Hospital in Bydgoszcz. The procedure was based on epidural implantation of the electrode at the level of Th1-Th2 and connection with the impulse generator located subcutaneously. As a result of the applied stimulation reduction of the anginal pain was achieved. PMID- 16273469 TI - [Osteoblastoma of the dens axis in a 14-year-old boy. A case report]. AB - We present a case of osteoblastoma of C1 in a 14-year-old boy. The case deserves attention because it is a conglomerate of issues important from the point of view of clinical practice. Key issues associated with this interdisciplinary pathology are discussed, such as diagnostic problems, stabilisation of the cervical spine and extensive surgery associated with the risk of severe bleeding in a Jehovah Witness. A short review of pertinent literature is included. PMID- 16273470 TI - [123I-metaiodobenzylguanidine myocardial scintigraphy in the differential diagnosis of patients with parkinsonian syndromes -- case reports]. AB - The aim of this study is to present a clinical role for 123I metaiodobenzylguanidine (MIBG) myocardial scintigraphy in the differential diagnosis in patients with parkinsonian syndromes. We present a 51-year-old woman with parkinsonian syndrome and syncope occurrence. She reproduced spontaneous syncope in the tilt test. We present also a 60-year-old man with parkinsonian syndrome and syncope and presyncope occurrence. He also reproduced spontaneous syncope in the tilt test. Cardiac 123I-MIBG scintigraphy was performed in both patients. In the former patient, the H/M ratio of MIBG uptake was within normal ranges, in the latter, it was abnormally impaired. The results of the 123I-MIBG cardiac scintigraphy confirm results of the other studies. In the patient with Parkinson's disease the H/M ratio of MIBG uptake was abnormally impaired. The patient with the multiple system atrophy was within normal ranges. PMID- 16273471 TI - The treatment of advanced chronic lower limb ischaemia with marrow stem cell autotransplantation. AB - INTRODUCTION: Conventional methods of critical leg ischaemia treatment are of limited efficacy. Amputation, as an ultimate solution, is not so rare. The results of marrow stem cell therapy as a potential novel approach to peripheral artery disease management were presented in 2002 by Tateishi-Yuyamy. AIM: To assess efficacy and safety of critical lower limb ischaemia treatment with marrow stem cell autotransplantation. METHODS: Ten patients suffering from chronic leg ischaemia in Fontaine IV stadium were involved in the study. They did not require emergency amputation and had previously been unsuccessfully treated with conventional therapy. Autologic marrow stem cells were condensated by a separator from bone marrow samples taken from the iliac crest. The cells were delivered intramuscularly by repeated injections into the pedal and tibial regions. The number of CD34 and AC133 positive mononuclear cells in each sample was evaluated by flow cytometry. After two weeks and one, two, three and twelve months the following parameters were measured: Laser Doppler Flux (LDF), percutaneous oxygen partial pressure, ankle-brachial index (ABI), visual analgesic scale (VAS), analgesic therapy requirement and ulceration area. Also, lower leg angiography and scintigraphy were performed. RESULTS: An improvement of the peripheral blood flow assessed by Laser Doppler Flux and percutaneous oxygen partial pressure was found. Pain severity decreased in the majority of patients. Amputation was required in three patients in whom the therapy failed. No side effects of the therapy were observed. The clinical effect of the treatment did not correlate with the amount of cells injected. CONCLUSION: Marrow stem cell autotransplantation into the ischaemic lower limb seems to be a potentially effective method of peripheral perfusion enhancement. Further studies are needed to clarify the underlying mechanisms of such improvement. PMID- 16273473 TI - Circumferential pulmonary vein RF ablation in the treatment of atrial fibrillation: 3-year experience of one centre. AB - INTRODUCTION: In patients with atrial fibrillation (AF), significantly symptomatic in particular, restoring and maintaining sinus rhythm is one of treatment strategies. Considering the limited efficacy and side effects of anti arrhythmic agents, growing hopes are attributed to the developing techniques of percutaneous ablation. AIM: To determine the efficacy and safety of circumferential pulmonary vein ablation performed using the CARTO system in patients with paroxysmal or permanent AF. METHODS: The study involved 94 patients (mean age 54 years, males 65%, structural heart disease 29.4%) with symptomatic, recurrent and AF resistant to antiarrhythmic agents (paroxysmal AF 63.8%), selected for circumferential pulmonary vein ablation with the Pappone method. Follow-up examinations were performed after 1, 3, 6, 9, and 12 months. The symptoms, ECG, 24-hour ECG monitoring and complications were recorded. RESULTS: Mean procedure and fluoroscopy durations were 4.5 hours and 22.4 minutes respectively. The long-term follow-up ranged from 3 to 24 months, with median time of 12 months. At six months, 47.8% of patients remained free from AF, and improvement in terms of infrequent arrhythmia occurrence and low incidence of symptoms in an additional 36.7% was observed. Efficacy was lower in patients with permanent AF (12 months 90% vs 70%). Complications were seen in six (6.4%) patients: cardiac tamponade in two patients; and pericardial effusion, retroperitoneal bleeding, stroke, and pulmonary vein thrombosis each in one patient. CONCLUSIONS: Circumferential pulmonary vein ablation leads to resolution of arrhythmia or marked clinical improvement in about 75% of patients with symptomatic, resistant AF. The success rate is lower in patients with permanent rather than paroxysmal AF. As severe complications are not unlikely, the indications for such therapy must be carefully balanced. PMID- 16273475 TI - The prognostic impact of renal failure in patients with ST-segment elevation acute myocardial infarction. AB - INTRODUCTION: Renal insufficiency (RI) is associated with higher morbidity and mortality in patients (P) with coronary artery disease and in P submitted to angioplasty. In ST-segment elevation acute myocardial infarction (STEAMI), this impact has not been well demonstrated. AIM: To evaluate the impact of RI in P with STEAMI. METHODS: We evaluated 160 P admitted with STEAMI, mean age of 62+/ 14 years, 76% male. We determined creatinine levels on admission. RI was defined as a level >1.5 mg/dl. Analysis of clinical, electrocardiographic and laboratory variables was performed, in relation to the endpoint defined as the occurrence of death at 30-day follow-up. RESULTS: There were 16 deaths (10%) at 30-day follow up. P with RI (n=21) were older (68+/-11 vs 61+/-14 years, p<0.001), more often had diabetes (57 vs 24 %, p=0.004) and presented more often with Killip class > or =2 (57 vs 12%, p<0.001). The use of statins (62 vs 83%, p=0.05) and beta blockers (24 vs 65%, p<0.001) was lower in P with RI. Mortality was higher in RI P (62 vs 2%, p<0.001). The univariate predictors of death were age > or =75 years, diabetes, Killip class > or =2 on admission, RI, non-use of statins and beta-blockers and use of diuretics. In multivariate analysis, independent predictors of death at 30 days were RI (HR 29.6, 95% CI 6.3-139.9, p<0.001) and non-use of beta-blockers (HR 0.13, 95% CI 0.02-1.01, p=0.01). CONCLUSION: In P admitted for STEAMI, the presence of RI was an independent predictor of death at 30 days whereas the usage of beta-blockers was protective. PMID- 16273477 TI - Evaluation of cerebral circulation in patients with significant carotid artery stenosis. AB - BACKGROUND: A significant proportion of ischaemic stroke episodes are caused by atherosclerotic lesions in extracranial arteries. Assessment of haemodynamical profile of cerebral arteries in both symptomatic and asymptomatic patients with carotid artery stenosis is of clinical importance. AIM: To assess haemodynamic changes in cerebral arteries in patients with significant internal carotid artery (ICA) stenosis. METHOD: Patients (n=109) were divided into the following groups: group I (GI) - 42 subjects (64.6+/-9.0 years) with asymptomatic ICA stenosis > or =70%; and group II (GII) - 67 subjects (63.4+/-7.1 years) after stroke. The control group consisted of 30 patients (60.3+/-8.9 years) without significant stenoses of extracranial arteries on USG and angiography. In all cases ultrasonographic evaluation of flow velocities and directions in cerebral arteries within the circle of Willis and collateral flow was performed. RESULTS: The severity of ICA stenosis did not differ significantly between GI and GII. Patients in GI had flow velocity in the middle cerebral artery (MCA) increased by 15.7% and by 40.8% in the anterior cerebral artery (ACA) contralateral to the ICA stenosis (p<0.001 and p<0.001), whereas in GII no significant changes in flow velocity in these arteries were observed in comparison with the control group. Patients in the groups I and II had lower flow velocities in MCA ipsilateral to the ICA stenosis, however values for GII patients were significantly lower than in GI patients (p<0.001). The presence of collateral circulation through the anterior and posterior communicating arteries (ACoA and PCoA) was similar in GI and GII; however, the flow velocities in the ipsilateral MCA and ACA were significantly higher in asymptomatic patients (GI). The frequency of active collateral circulation through both ACoA and PCoA increased along with the increase of ICA stenosis severity (p=0.003; p<0.001). CONCLUSIONS: Collateral flow in the circle of Willis in subjects with ICA stenosis occurs equally often in symptomatic and asymptomatic patients; however, it is more efficient in patients without symptoms. The rate of development of collateral circulation depends on ICA stenosis severity. The important role in maintaining collaterals within the circle of Willis is played by ACoA, although in some patients MCA may also be supplied by PCoA. PMID- 16273479 TI - Implantable cardioverter-defibrillators in patients with hypertrophic cardiomyopathy -- dilemmas and difficulties. AB - INTRODUCTION: The implantation of a cardioverter-defibrillator (ICD) is an established method of sudden cardiac death (SCD) prevention. The value of ICD therapy in secondary prevention of SCD is unquestionable. Precise identification of high-risk patients and ICD use for primary prevention of SCD, especially in patients with hypertrophic cardiomyopathy (HCM), remain controversial. Problems include the high prevalence of complications associated with ICD implantation and optimal selection of ICDs. AIM: To estimate the frequency and type of complications after ICD implantations in HCM patients in a long-term follow-up. METHOD: The efficacy and safety of ICD therapy were estimated in 46 HCM patients with devices implanted for a secondary (n-18) or primary prevention (n-28) of SCD. RESULTS: During the mean follow-up period of 28.2+/-26.1 months (from 2 to 68) appropriate ICD interventions occurred in 10 (55%) patients of the secondary prevention group and in 3 (10%) patients of the primary prevention group. Complications were documented in 15 (33%) patients. The most frequent were inappropriate ICD interventions recorded in 14 (30%) patients. The causes of these inappropriate ICD shocks were: T-wave oversensing (7 patients), atrial fibrillation with rapid ventricular rhythm (3 patients), lead failure (2 patients), and sinus tachycardia (2 patients). In two patients infections of the ICD pocket requiring removal of the system occurred. Displacement of the lead occurred in one patient. There were no significant differences in the prevalence of complications between the primary and secondary prevention groups or in the number of inappropriate interventions with respect to ICD type. CONCLUSIONS: The high rate of appropriate ICD shocks provides proof of high ICD-based SCD prevention efficacy. There is a high rate of complications observed after ICD implantation with inappropriate interventions being the most frequent among them. This indicates that careful programming of the device as well as the use of a programme with T-wave oversensing prevention should be ensured. PMID- 16273481 TI - One-year clinical outcome after emergency hospitalisation for suspected acute coronary syndrome -- a comparative analysis with respect to angiographic findings. AB - INTRODUCTION: Among patients with suspected acute coronary syndromes (ACS) referred for urgent coronary angiography there are some with normal angiograms. AIM: To compare, with respect to angiographic findings, one-year clinical outcomes in patients hospitalised for suspected ACS. METHOD: Between January 2001 and December 2003 emergency angiography was performed in 1169 patients. It revealed no significant coronary lesions in 97 (8.3%) cases, of these 40 being women and 57 men with a mean age of 55+/-15 yrs (Group 1). Sixty consecutive patients with ACS (20 women and 40 men with a mean age of 54+/-9 yrs) and significant coronary artery disease (CAD) confirmed by angiography were the study controls (Group 2). Demographic data, CAD risk factors, angiographic findings, ECG and echocardiographic data as well as laboratory test results were analysed. In a long-term follow-up, the prevalence of recurrent angina and all-cause mortality were assessed. RESULTS: There were no significant differences in patients' demographics between the two groups with the exception of arterial hypertension, which was more frequent in group 1. In group 1 the diagnosis of ACS was established in 14.5% cases, X syndrome in 14.4%, Prinzmetal angina in 4.1%, myocarditis in 6.1% and pulmonary thromboembolization in 5.1%. A definite diagnosis was not related to the cardiovascular disease in 36%. No complications were observed in the late follow-up of group 1 patients. CONCLUSIONS: Apparently normal angiograms are relatively common in patients referred for emergency coronary angiography. In patients without significant CAD on their angiograms the clinical outcome is favourable, without major adverse cardiovascular events in the long-term follow-up. PMID- 16273482 TI - [The influence of deterioration of clinical condition on the change of revascularisation method in coronary artery disease]. AB - A case of a 52-year old woman with diabetes mellitus and symptomatic, multivessel coronary disease is presented. The patient underwent coronary angiography. Taking in consideration the intensity of atherosclerotic lesions and vessels diameter, she was initially selected for cardiovascular surgery. However, because of sudden deterioration of clinical condition caused by myocardial infarction the strategy of cardiac revascularisation was changed. The patient underwent multivessel percutaneous coronary intervention with good result in short- and 6-month observation. PMID- 16273483 TI - [Thrombotic thrombocytopenic purpura -- a rare complication associated with clopidogrel therapy. A case report]. AB - Clopidogrel -- a popular antiplatelet drug widely used in treatment of various vascular diseases -- is known for its favourable safety profile. Thrombotic thrombocytopenic purpura (TTP) is very rare but serious adverse reaction associated with clopidogrel. We present a case of patient treated with clopidogrel who developed moderate TTP with leukopenia and symptoms of haemorrhagic diathesis. PMID- 16273484 TI - [Infective endocarditis in a patient with multiple myeloma. A case report]. AB - A case of a 59 year old male with infective endocarditis is presented. Antibiotic therapy seemed effective, however, inflammation laboratory parameters increased two weeks after clinical improvement and body temperature normalisation. Subsequent extensive laboratory investigations revealed multiple myeloma. The patient underwent successful aortic valve replacement and received pharmacological therapy for multiple myeloma. Difficulties in diagnosing and treatment of patients with infective endocarditis who have other concomitant diseases, are discussed. PMID- 16273485 TI - [Congestive heart failure after the Senning operation. Case report]. AB - From the early 1960s to the mid 1980s, the Mustard and Senning procedures were the treatment of choice for transposition of the great arteries (d-TGA). We report a case of a young girl who had undergone surgical repair of complete transposition by the Senning procedure in the early infancy. Twelve years later she developed a congestive heart failure. Based on the x-ray, echo and angiography study she was qualified for reoperation at German Heart Institute Berlin with a very good result. Long term follow-up of patients who underwent Mustard or Senning repair for d-TGA leads to identification of multiple residua and sequelae, as well as functional problems associated with these two procedures. Our patient illustrates the effective therapy which can improve the quality of life and postpone the time for the heart transplantation. PMID- 16273486 TI - [Palliative stenting of venous graft in a patient with multivessel coronary artery disease]. PMID- 16273487 TI - [Tachycardia with rSr' <110 ms in lead V1 -- is it always an atrio-ventricular nodal tachycardia?]. AB - We present a case of a 15-year old girl in whom supraventricular tachycardia (SVT) with rSr' pattern in lead V1 and the QRS width of less than 110 ms, suggesting typical slow-fast AVNRT, was the presenting arrhythmia. During sinus rhythm no preexcitation was observed. Considerable variability of the tachycardia cycle length (240-370 ms), attributable to the presence of the fast and slow nodal pathways, was also observed. However, during the electrophysiological study only orthodromic atrio-ventricular reciprocating tachycardia with the left-sided superior accessory pathway as the retrograde limb was documented. Thus, rSr' pattern was due to incomplete right bundle branch block. We describe how the correct diagnosis and the site of the atrial insertion of the accessory pathway could be inferred from the careful analysis of the P' wave morphology during the tachycardia. PMID- 16273489 TI - [ST segment elevation after pacemaker implantation]. PMID- 16273488 TI - [Thrombus visualisation during radiofrequency catheter ablation. A case report]. AB - We report two patients in whom thrombus formation during radiofrequency catheter ablation was detected by echocardiography. Resolution of thrombus after intravenous use of heparin was observed in both patients. Transesophageal and intracardiac echocardiography may be useful in management of this complication. PMID- 16273490 TI - [ASCOT Study confirms the benefit of new classes of hypotensive drugs]. PMID- 16273491 TI - [Left main coronary artery disease -- only for cardiac surgeon?]. PMID- 16273492 TI - Interpretation of clopidogrel resistance. PMID- 16273495 TI - [Pain treatment of patients with incurable malignant tumors]. AB - BACKGROUND AND OBJECTIVE: Efficacious pain therapy is one of the main challenges in the management of patients with advanced cancer. It was the aim of this study to ascertain whether adequate pain treatment was achieved in a palliative care unit and what changes in medical treatment were undertaken during the patients stay in this unit. SUBJECTS AND METHODS: Medication against pain and pain intensity were documented during hospital care of 94 patients with cancer (mean age: 66.8 years; female/male 52.2/47.8%; discharged/deceased 58.5/41.5%) at admission and discharge. 32 patients were interviewed by phone after discharge about their current perception and intensity of pain, as well as their health. RESULTS: Highly significant reduction of pain intensity was achieved in all patients while receiving palliative care. Only those patients who were on oral morphine at admission to hospital required significantly raised morphine dosages while receiving palliative treatment. The other patients did not need significant changes of opioid medication. Pain reduction persisted even after discharge. CONCLUSION: In patients with advanced cancer the effects of psychological, social and spiritual factors on their perception of pain is probably of greater importance in achieving efficacious pain reduction than a change of medication. Further studies will be needed to confirm these findings. PMID- 16273494 TI - [Driving simulator performance in patients with obstructive sleep apnea syndrome: what consequences for driving capability?]. AB - BACKGROUND AND OBJECTIVE: Patients with obstructive sleep apnea (OSA) have an increased accident risk. The German Society of Sleep Research and Sleep Medicine (DGSM) recommends for patients with OSA and daytime sleepiness that their driving ability should be re-established 6 weeks after the initiation of CPAP (continuous positive airway pressure), with documentation of therapeutic effects on daytime symptoms and performance. The present study was conducted to investigate whether an improvement of driving ability can be documented in neuropsychological tests and a simulated driving situation 14 days after the initiation of CPAP. PATIENTS AND METHODS: Driving simulation and neuropsychological tests of vigilance were conducted in 36 patients (36 males, aged 54 9 years) with OSAS before and 2 (n=23), 14 (n=18) and 42 days (n=17) after initiation of CPAP. RESULTS: Vigilance tests showed only slight changes under CPAP. Frequency of accidents during driving simulation was reduced after 14 days of CPAP, but a statistically remarkable decrease was achieved only on day 42. In contrast, concentration faults were reduced after 2 and 14 days of CPAP. CONCLUSIONS: In OSA-patients improvement of daytime performance in a simulated driving situation can be documented 14 days after initiation of CPAP. There is need for investigating larger cohorts of patients so that current recommendations for driving licensing can be modified and permission to drive can be given earlier. PMID- 16273496 TI - [Human infection with Gongylonema pulchrum]. AB - HISTORY: A 27-year-old woman repeatedly attended an outpatient clinic and reported sensations of movement within the buccal mucosa. Various local inspections were without any obvious abnormal findings. A few days later the patient brought a filiform structure, which she had extracted from the affected mucosa by manipulation with her toothbrush. INVESTIGATION: Microscopy indicated a gravid female nematode of the genus Gonglyonema. DIAGNOSIS: Gonglyonem pulchrum infection. TREATMENT AND COURSE: Careful inspection revealed a second worm at the other side of the oral cavity, which was extracted by local incision. After removal of the second worm the patient remained free of symptoms without any further therapy. CONCLUSION: Even obviously very unusual symptoms, which might be indicative of a dermatozoic delusion, should be carefully clarified. In some cases it may be appropriate to consult a specialized institutions or laboratory to rule out a possible parasitic infections. PMID- 16273497 TI - [Cor pulmonale and atrial thrombus]. PMID- 16273501 TI - [Treatment of proven growth hormone deficiency in adults with recombinant human growth hormone according to evidence-based criteria]. PMID- 16273502 TI - [Use of pharmaceuticals after the expiration date?]. PMID- 16273498 TI - [Nephrotoxic drugs]. PMID- 16273504 TI - [Is there a German precedent for evidence-based medicine?]]. PMID- 16273505 TI - Computation of vertical excitation energies of retinal and analogs: scope and limitations. AB - A comprehensive survey of computational methods: semiempirical (ZINDO/S), Time Dependent Hartree-Fock (TD-HF), Configuration Interaction Singles (CIS), and several approximate functionals within the Time-Dependent Density Functional Theory (TD-DFT) has been carried out for the description of vertical excitation energies and oscillator strengths of retinal and related polyenals. ZINDO and TD DFT computations showed the best agreement with the experimental data. In particular, hybrid functionals including approximately 25% of exact exchange (B3LYP, B3P86, and PBE0) were found to perform best with these highly conjugated polyenes. A systematic average error of 0.18-0.22 eV has been found after a simple one-parameter correction. Thus, 0.18 eV might be considered the upper limit of accuracy for current one-determinant methods in the computation of vertical excitation energies. The consideration of adiabatic excitations, conformational sampling, solvation, and nondynamic correlation should describe this processes more accurately, but this leads to highly demanding methods beyond feasibility for these large polyenes. The trends observed, particularly the good performance of the ZINDO/S method, should pave the way for the prediction of excited states properties in natural and artificial photoreceptor proteins, thus advancing towards the description of their light-transducing biological role in Nature. PMID- 16273506 TI - A charge-scaling implementation of the variational electrostatic projection method. AB - Two new charge-scaling methods for efficient modeling of the solvated macromolecular environment in hybrid QM/MM calculations of biological reactions are presented. The methods are extensions of the variational electrostatic projection (VEP) method, and allows a subset of atomic charges in the external environment to be adjusted to mimic, in the active dynamical region, the electrostatic potential and field due to the large surrounding macromolecule and solvent. The method has the advantages that it offers improved accuracy, does not require the use of a three-dimensional grid or auxiliary set of fitting points, and requires only minor molecular simulation code modifications. The VEP-cs and VEP-RVM+cs methods are able to attain very high accuracy (relative force errors of 10(-7) or better with appropriate choice of control parameters), and take advantage of a recently introduced set of high-order discretization schemes and Gaussian exponents for boundary element solvation and VEP methods. The methods developed here serve as potentially powerful tools in the arsenal of computational techniques used in multiscale computational modeling problems. PMID- 16273507 TI - Is anoxic depolarisation associated with an ADC threshold? A Markov chain Monte Carlo analysis. AB - A Bayesian nonlinear hierarchical random coefficients model was used in a reanalysis of a previously published longitudinal study of the extracellular direct current (DC)-potential and apparent diffusion coefficient (ADC) responses to focal ischaemia. The main purpose was to examine the data for evidence of an ADC threshold for anoxic depolarisation. A Markov chain Monte Carlo simulation approach was adopted. The Metropolis algorithm was used to generate three parallel Markov chains and thus obtain a sampled posterior probability distribution for each of the DC-potential and ADC model parameters, together with a number of derived parameters. The latter were used in a subsequent threshold analysis. The analysis provided no evidence indicating a consistent and reproducible ADC threshold for anoxic depolarisation. PMID- 16273509 TI - In vivo single-shot, proton-localized 13C MRS of rhesus monkey brain. AB - A single-shot, proton-localized, polarization transfer (13)C spectroscopic method was proposed and implemented on a 4.7 T scanner for studying rhesus monkey brains. The polarization transfer sequence was mostly adiabatic, minimizing signal loss due to B(1) inhomogeneity. RF pulses in polarization transfer were also used for voxel selection of protons with gradient fields. The transferred (13)C magnetization was refocused by additional refocusing adiabatic pulses. With the intravenous infusion of D-[1-(13)C]glucose solution, (13)C NMR spectra from a 30 mL voxel were acquired for the resonances of C1 of glucose, C2,3,4 of glutamate and glutamine. The time-resolved turnover of glutamate, glutamine and aspartate from intravenously infused D-[1-(13)C]glucose at a temporal resolution of 12 min was demonstrated with excellent spectral resolution and signal-to-noise ratio. Typically, the half-height linewidth of the decoupled (13)C peaks was approximately 4 Hz. Data obtained with infusion of sodium [2-(13)C]acetate using the proposed polarization transfer method and data from the carboxylic carbon region using non-localized acquisition are also presented. PMID- 16273508 TI - Grey and white matter GABA level differences in the human brain using two dimensional, J-resolved spectroscopic imaging. AB - A novel, two-dimensional, J-resolved chemical-shift imaging sequence was used to collect gamma-aminobutyric acid (GABA) spectroscopic imaging data on six healthy subjects at 4 T. Using image segmentation and a linear-regression analysis relating brain GABA level to tissue-type, a consistent and significant (n = 6, p < 0.01) elevation of mean GABA levels was measured in the cortical grey matter (0.96 +/- 0.24 mm) compared with white matter (0.44 +/- 0.16 mm) across all six subjects. The results suggest an approximately two-fold elevation of GABA levels in cortical grey matter compared with white matter in vivo. Our findings are consistent with ex vivo studies in the literature of both animal and human brain and demonstrate the significant potential of this technique for detecting and quantifying tissue-specific neurochemical pathology in vivo. PMID- 16273510 TI - Germ cell lineage differentiation in non-seminomatous germ cell tumours. AB - Human germ cell tumours (GCTs) have long fascinated investigators for a number of reasons. Being pluripotential tumours, they can differentiate into both extra embryonic and embryonic (somatic) tissues. However, it has never been shown convincingly that, in humans, these tumours are truly totipotent and can also give rise to the germ lineage, the third major differentiation lineage occurring early during embryonic life. Using a number of newly available, distinct, immunohistochemical markers, such as OCT3/4, VASA and TSPY, the occurrence of germ cells was investigated in a number of germ cell tumours. Development of germ cells was identified in three independent non-seminomas, including two pure yolk sac tumours and one mixed tumour composed of yolk sac tumour and immature teratoma. Our finding indicates a previously unknown totipotent potential of human GCTs and raises the question of whether, under certain culture conditions, primordial germ cells could be derived from human GCT cell lines. PMID- 16273511 TI - Ethical considerations in writing psychological assessment reports. AB - In this article, the author addresses the ethical questions and decision evaluators associated with the writing of psychological assessment reports. Issues related to confidentiality, clinical judgment, harm, labeling, release of test data, and computer usage are addressed. Specific suggestions on how to deal with ethical concerns when writing reports are discussed, as well as areas in need of further research. PMID- 16273512 TI - Forensic report writing. AB - As the area of specialization of forensic psychology broadens its horizons, it will become increasingly more difficult for clinical psychologists to avoid participating in the forensic arena. As a result, clinical psychologists may be asked to write reports about therapeutic intervention or evaluations in divorce, personal injury, competency, abuse, or criminal proceedings. In this article, the author addresses the issues that the clinical psychologist needs to understand in writing reports that may be utilized in forensic settings. How these reports are affected by the American Psychological Association Ethics Code (APA; 2002), the audience receiving the report, the subject matter of the report, and dissemination of the information are all discussed in this article. PMID- 16273513 TI - The psychological report: a review of current controversies. AB - A summary of the most frequent controversies in report writing is covered. These include length, readability, acknowledging use of poorly validated measures, use of computer-based narratives, inclusion of test scores, degree of integration, inclusion of client strengths, and development of a feedback report. Available research is summarized along with suggestions for future research. PMID- 16273514 TI - Conformation of alloHyp in the Y position in the host-guest peptide with the pro pro-gly sequence: implication of the destabilization of (Pro-alloHyp-Gly)10. AB - The crystal structure of the host-guest peptide, (Pro-Pro-Gly)4-(Pro-alloHyp-Gly) (Pro-Pro-Gly)4, was analyzed at high resolution. allohydroxyproline (alloHyp), 4S hydroxyproline, was successfully characterized through the use of a host-guest peptide, while the previous study indicated the inability of a triple helical formation of (Pro-alloHyp-Gly)10. A detailed analysis of alloHyp conformation in collagen-like models sheds light on the role played by its puckering in the triple-helix stabilization and destabilization. That is, the alloHyp typically adopts down puckering. However, it adopted up puckering in the Y position in the Pro-alloHyp-Gly guest triplet, which was not preferable conformation for alloHyp. Therefore, the energetically unfavorable conformations seemed to play the key role in giving destabilization to the triple helix in (Pro-alloHyp-Gly)10. The intrinsic hydration pattern in (Pro-Pro-Gly)9 was conserved even in the surrounding alloHyp residues. PMID- 16273515 TI - Thermal stabilization of human albumin by medium- and short-chain n-alkyl fatty acid anions. AB - A comprehensive study of the thermal stabilization of defatted human albumin monomer by n-alkyl fatty acid anions (FAAs), formate through n-decanoate, was carried out by differential scanning calorimetry (DSC). The concentration of each ligand affording maximum thermal stabilization was determined; n-nonanoate provides the greatest stabilization but is only marginally better than n octanoate and n-decanoate. The use of reversible thermodynamics and a two-state denaturation model for albumin has been validated. Standard free energies of binding, calculated from increases in free energy of denaturation, for n butanoate and longer FAAs, are linear with n-alkyl chain length whereas those for formate, acetate, and n-propionate deviate from linearity; those for acetate and n-propionate are even greater than that of n-butanoate, thereby suggesting, in addition to the common class of sites available to all such ligands, the presence of an additional class of lower affinity binding sites available only to these shortest ligands. Competition experiments involving acetate and n-octanoate and involving n-pentanoate and n-octanoate confirmed the binding of acetate to lower affinity sites unavailable to n-octanoate and n-pentanoate. Furthermore, an equation is provided, allowing computation of the transition temperature as a function of the free energy for any reversible process causing a change in thermal stability of a protein undergoing reversible, two-state denaturation. With this equation, modeling the competition experiments by using the binding parameters determined by DSC provides additional support for the class of lower affinity sites, which play a significant role in thermal stabilization of albumin at higher concentrations of these shortest FAAs. PMID- 16273516 TI - Variables affecting the clarity of psychological reports. AB - Effective psychological reports are consumer-focused: They address the concerns of the referring persons, present data appropriately, communicate clearly and concisely, and include useful and appropriate recommendations. Although the importance of clear communication has been stressed repeatedly, psychologists often write reports that are very difficult for nonpsychologists to read. In this article, the author explores four reasons behind this dichotomy: (a) model reports available to psychologists in training are written at a level that is very difficult to understand; (b) psychological terms are not commonly defined; (c) the amount of time it takes to write easily understood reports is substantial; and (d) psychologists are confused about how to address multiple audiences. Methods to address each issue are discussed. PMID- 16273517 TI - Introduction to the special series on psychological reports. PMID- 16273518 TI - Computer utilization and clinical judgment in psychological assessment reports. AB - The process of assessment report writing is a complex one, involving both the statistical evaluation of data and clinical methods of data interpretation to appropriately answer referral questions. Today, a computer often analyzes data generated in a psychological assessment, at least in part. In this article, the author focuses on the interaction between the decision-making processes of human clinicians and the test interpretations that are computer-based. The benefits and problems with computers in assessment are highlighted and are presented alongside the research on the validity of automated assessment, as well as research comparing clinicians and computers in the decision-making process. The author concludes that clinical judgment and computer-based test interpretation each have weaknesses. However, by using certain strategies to reduce clinicians' susceptibility to errors in decision making and to ensure that only valid computer-based test interpretations are used, clinicians can optimize the accuracy of conclusions that they draw in their assessment report. PMID- 16273519 TI - Balancing psychological assessments: including strengths and hope in client reports. AB - Borrowing from B. A. Wright's (1991) suggestions, a balanced approach is advocated for gathering information and writing the subsequent report. Specifically, it is suggested that the clinician attend to four aspects of an interviewee: (a) the strengths in the client's psychological makeup, (b) the weaknesses in the client's psychological makeup, (c) the strengths in the client's environment, and (d) the weaknesses in the client's environment. Additionally, using hope theory (C. R. Snyder, 1994) as a framework, the importance of including information about client goals, along with the routes to those goals (pathways thinking) and the motivation to use those pathways (agency thinking), is described. Furthermore, the implications of using this framework in conducting a diagnostic interview and writing the ensuing report are provided. Finally, the advantages of including human strengths to achieve a balanced interview and report are discussed. PMID- 16273520 TI - Migratory routes and fates of cells transcribing the Wnt-1 gene in the murine hindbrain. AB - To investigate the origins, migrations, and fates of Wnt-1-expressing cells in the murine hindbrain, mice carrying a Wnt-1 enhancer/lacZ transgene were observed from embryonic day (E) 8 through postnatal day 18. The transgene-stained ventricular layer waxed and waned prior to and following migrations from it. Stained cells migrated first external to the hindbrain as neural crest and then within it to form typical populations of the rhombic lip, as well as others not recognized as lip derivatives. Migrations originated in a temporally defined sequence, many from discrete rhombomeres. All moved first radially, then rostrally and/or ventrally, ipsi-, or contralaterally, in the mantle or marginal layers. These movements ultimately formed elements of several nuclei, aligned in four longitudinal bands: dorsal (including the gracile, cuneate, cochlear, and vestibular nuclei, plus cerebellar granular cells), dorsal intermediate (including trigeminal sensory, parvicellular reticular, and deep cerebellar nuclei), ventral intermediate (including lateral and intermediate reticular nuclei), and ventral (including the raphe obscurus and pontine nuclei). Transgene staining often persisted long enough to identify stained cells in their definitive, adult nuclei. However, staining was transient. The strength of the staining, however, was in its ability to reveal origins and migrations in both whole-mounts and sections, in single cell detail. The present results will permit analyses of the effects of genetic manipulations on Wnt-1 lineage cells. PMID- 16273521 TI - Imbalance in liver homeostasis leading to hyperplasia by overexpressing either one of the Bcl-2-related genes, zfBLP1 and zfMcl-1a. AB - Apoptosis is an essential part of normal embryonic development in vertebrates, and it is involved in sculpturing organs and controlling cell populations. In previous studies, we identified two novel proteins, zfBLP1 and zfMcl-1a, which are similar to those of the Bcl-2 family as a group of evolutionarily conserved proteins that regulate cellular anti-apoptosis. To evaluate the effect of dysregulated hepatocyte apoptosis during zebrafish hepatogenesis, we demonstrate the transgenic overexpression of either zfBLP1 or zfMcl-1a in zebrafish larval liver. Results showed that 18%-43% of larvae overexpressed zfBLP1 and that 16% 37% of larvae overexpressed zfMc1-1a in the liver leading to liver hyperplasia in 5-day postfertilization (dpf) zebrafish larvae. Histologically, zebrafish larvae exhibiting liver hyperplasia displayed a normal type of hepatocyte and the same cell numbers in their two liver buds compared with only one liver bud of wild type larvae. Of interest, the expression of cyclin genes (A2, B, D1, and E), hepatocyte nuclear factor genes (HNF-1alpha, beta, -3beta, and 4alpha), and oncogenic markers (P53, c-myc, beta-catenin, N-ras, and gankyrin) were up regulated, while the expression of C/EBP-alpha was down-regulated in a zfMcl-1a mediated anti-apoptotic process of the liver. Increased cell death and proliferation was found in both hepatic cells of zebrafish larvae overexpressing either zfBLP1 or zfMcl-1a. However, those zebrafish larvae with liver hyperplasia only lived approximately 10 days. (This finding may have been due to liver abnormalities that led to failure of liver function.) In conclusion, transgenic overexpression of zfBLP1 or zfMcl-1a in zebrafish larvae interrupts regulation of the homeostatic balance between cell proliferation and programmed cell death during hepatogenesis and leads to liver hyperplasia. PMID- 16273522 TI - Discrete pulses of molting hormone, 20-hydroxyecdysone, during late larval development of Drosophila melanogaster: correlations with changes in gene activity. AB - Periodic pulses of the insect steroid molting hormone 20-hydroxyecdysone (20E), acting via its nuclear receptor complex (EcR/USP), control gene expression at many stages throughout Drosophila development. However, during the last larval instar of some lepidopteran insects, subtle changes in titers of ecdysteroids have been documented, including the so-called "commitment peak." This small elevation of 20E reprograms the larva for metamorphosis to the pupa. Similar periods of ecdysteroid immunoreactivity have been observed during the last larval instar of Drosophila. However, due to low amplitude and short duration, along with small body size and staging difficulties, their timing and ecdysteroid composition have remained uncertain. Employing a rigorous regimen of Drosophila culture and a salivary gland reporter gene, Sgs3-GFP, we used RP-HPLC and differential ecdysteroid RIA analysis to determine whole body titers of 20E during the last larval instar. Three small peaks of 20E were observed at 8, 20, and 28 hr following ecdysis, prior to the well-characterized large peak around the time of pupariation. The possible regulation of 20E levels by biosynthetic P450 enzymes and the roles of these early peaks in coordinating gene expression and late larval development are discussed. PMID- 16273523 TI - Inhibition of zebrafish fin regeneration using in vivo electroporation of morpholinos against fgfr1 and msxb. AB - Increased interest in using zebrafish as a model organism has led to a resurgence of fin regeneration studies. This has allowed for the identification of a large number of gene families, including signaling molecules and transcription factors, which are expressed during regeneration. However, in cases where no specific inhibitor is available for the gene product of interest, determination of a functional role for these genes has been difficult. Here we demonstrate that in vivo electroporation of morpholino oligonucleotides is a feasible approach for protein knock-down during fin regeneration. Morpholino oligonucleotides against fgfr1 and msxb were utilized and knock-down of both proteins resulted in reduced fin outgrowth. Importantly, Fgfr1 knock-down phenocopied outgrowth inhibition obtained with an Fgfr1 inhibitor. Furthermore, this method provided direct evidence for a functional role for msxb in caudal fin regeneration. Finally, knock-down of Fgfr1, but not Msxb, affected the blastemal expression of msxc, suggesting this technique can be used to determine epistasis in genetic pathways affecting regeneration. Thus, this convenient reverse genetic approach allows researchers to quickly (1) assess the function of genes known to be expressed during fin regeneration, (2) screen genes for functional relevance during fin regeneration, and (3) assign genes to the molecular pathways underlying fin regeneration. PMID- 16273525 TI - Hot hearts in the Sonoran Desert: the 11th Weinstein Cardiovascular Development Conference in Tucson. AB - The 11th Annual Weinstein Cardiovascular Development Conference was held May 19 22, 2005 at the Westward Look Resort and Conference Center in Tucson, Arizona. The Westward Look was the site of the 6th Weinstein Meeting in 1999, and this year, 330 basic research scientists and research clinicians returned to Tucson for 3 days of meetings, Mariachis, and margaritas. The meeting was hosted by the cardiovascular research group at the University of Arizona and offered flavors of the desert southwest that included record temperatures, the Skopopelli conference logo modified from the Kokopelli of Native American mythology (Fig. 1), and liberal use of a cattle prod to encourage speaker timeliness. PMID- 16273524 TI - Pea3 expression is regulated by FGF signaling in developing retina. AB - FGF signaling has been implicated as an important regulator of retinal development. As a first step in characterizing potential downstream targets of FGF signaling in the retina, we have analyzed expression of Pea3, a member of the Pea3 class of Ets-domain transcription factors, in the developing eye. We find that Pea3 is expressed in the developing retina, and its transcription is regulated by FGF receptor activation. In addition, FGF signaling activates Cath5, a gene necessary for retinal ganglion cell differentiation. These results suggest that FGF signaling via MAPK up-regulates transcription factors that in turn control retinal ganglion cell differentiation. PMID- 16273526 TI - One- and two-photon induced fluorescence of Pacific Blue-labeled human serum albumin deposited on different core size silver colloids. AB - We studied one- and two-photon induced fluorescence of Pacific Blue (PB)-labeled human serum albumin (HSA) in the presence of different size silver colloids. The PB fluorescence emission intensity was observed with small (30-40 nm) and large (about 120 nm) colloids and compared with PB emission in absence of colloids. For the system with a small core size colloids we did not detect any fluorescence enhancement with one-photon excitation and the enhancement observed with two photon excitation was about 2.5-fold. In contrast, for large silver colloids we observed about a 2-fold increase in PB fluorescence brightness for one-photon excitation, and the enhancement with two-photon excitation excided 13-folds. Much stronger increases in brightness observed with two-photon excitation, compared to one-photon excitation, indicate a dominant role of enhanced local field in fluorescence enhancement on silver colloids in solutions. PMID- 16273527 TI - Expression of chondrogenic potential of mouse trunk neural crest cells by FGF2 treatment. AB - There is a significant difference between the developmental patterns of cranial and trunk neural crest cells in the amniote. Thus, whereas cranial neural crest cells generate bone and cartilage, trunk neural crest cells do not contribute to skeletal derivatives. We examined whether mouse trunk neural crest cells can undergo chondrogenesis to analyze how the difference between the developmental patterns of cranial and trunk neural crest cells arises. Our present data demonstrate that mouse trunk neural crest cells have chondrogenic potential and that fibroblast growth factor (FGF) 2 is an inducing factor for their chondrogenesis in vitro. FGF2 altered the expression patterns of Hox9 genes and Id2, a cranial neural crest cell marker. These results suggest that environmental cues may play essential roles in generating the difference between developmental patterns of cranial and trunk neural crest cells. PMID- 16273528 TI - Anorectal motility responses to selective stimulation of the ventral sacral nerve roots in an experimental model. AB - BACKGROUND: Control of defaecation and continence may be lost in patients with spinal cord injury. Electrical stimulation of sacral nerve roots to promote defaecation simultaneously activates both the rectum and the external anal sphincter (EAS), and may actually obstruct defaecation. The aim of this study was to investigate whether the EAS could be blocked selectively by selective stimulation of the ventral sacral nerve roots, and whether activation of the rectum without activation of the EAS could be obtained by stimulation of the ventral sacral nerve roots. METHODS: Selective electrical stimulation was performed using anodal blocking, a tripolar cuff electrode and monophasic rectangular current pulses applied to the sacral nerve roots in nine Gottingen minipigs. RESULTS: Simultaneous responses in the rectum and the anal canal were observed in five animals, whereas only anal responses were noted in four. Variations in cross-sectional area and an increase in rectal pressure seemed to facilitate defaecation. Without blocking, the increase in anal canal pressure was 16-45 cmH(2)O. With blocking, this increase was abolished in seven and reduced to 3-6 cmH(2)O in two animals. CONCLUSION: Selective activation of the rectum is possible using an anodal block of somatic motor fibres. This technique holds promise in further development of electro-defaecation. PMID- 16273529 TI - Operative mortality after colorectal resection in the Netherlands. AB - BACKGROUND: The aim of this study was to quantify factors related to operative mortality after colorectal resection in the Netherlands. METHODS: Multilevel logistic regression modelling was used. Institutional effects were calculated with and without adjustment for specific patient (age, sex, urgency of operation) and hospital (number of procedures, type of hospital) characteristics. All adult Dutch patients who underwent primary colorectal resection between 1994 and 1999 were included, except those who had (sub)total colectomy or local rectal resection. RESULTS: A total of 67 594 patients underwent colorectal resection. The in-hospital mortality rate was 7.0 per cent (elective 3.9 per cent, acute 14.3 per cent). Acute operation (odds ratio 3.89) and age (odds ratios 2.63, 5.23 and 10.13 for patients aged 50-69, 70-79 and 80 or more years respectively compared with those aged less than 50 years) had the strongest effects, followed by male sex (odds ratio 1.48) and type of hospital. There was no difference in operative mortality rate between low-, medium- and high-volume hospitals. CONCLUSION: In the Netherlands, advanced age and acute operation are by far the most important factors related to operative mortality after colorectal resection. Male sex and type of hospital have only a modest effect, and there is no discernible effect of hospital volume. PMID- 16273530 TI - The evolving role of gene-based treatment in surgery. AB - BACKGROUND: The completion of the sequencing of the human genome in 2003 marked the dawn of a new era of human biology and medicine. Although these remarkable scientific advances improve the understanding of human biology, the question remains how this rapidly expanding knowledge of functional genomics affects the role of surgeons. This article reviews the potential therapeutic application of gene therapy for various surgical conditions. METHODS: The core of this review was derived from a Medline database literature search. RESULTS AND CONCLUSION: The currently available vectors in the field of gene therapy and their limitations for clinical applications were analysed. The achievements of gene therapy in clinical trials and the future ramifications for surgery were also explored. Whether gene therapy takes a major role in surgical practice will depend greatly on the success of future vector development. Advances in viral vector technology to reduce the inflammatory effect, and improvements in the efficiency of gene delivery using non-viral vector technology, would allow this form of therapy to become more clinically applicable. PMID- 16273531 TI - Laparoscopic liver resection. AB - BACKGROUND: This paper describes a 10-year experience of laparoscopic liver surgery, including several major hepatectomies for malignant tumours. METHODS: Of 243 hepatectomies carried out between January 1995 and December 2004, 113 (46.5 per cent) were performed by laparoscopy and 89 were included in this retrospective study. RESULTS: Twenty-four laparoscopic hepatectomies (27 per cent) were for benign disease and 65 (73 per cent) for malignant tumours, including hepatocellular carcinoma (HCC) in 16 patients and colorectal metastasis (CRM) in 41. Minor hepatectomy was performed in 51 patients and major hepatectomy (three or more Couinaud segments) in 38. Conversion to laparotomy was necessary in 12 patients and perioperative blood transfusion in eight. One patient with cirrhosis who underwent right hepatectomy for HCC with conversion to open surgery died 8 days after surgery. Major morbidity occurred in eight patients (16 per cent) having minor hepatectomy and in 11 (29 per cent) of those having a major resection. The 3-year overall and disease-free survival rates for patients with CRM (mean follow-up 30 months) were 87 (11 patients at risk) and 51 (6 patients at risk) per cent respectively. Corresponding values for patients with HCC (mean follow-up 40 months) were 85 (10 patients at risk) and 68 (5 patients at risk) per cent. CONCLUSION: In experienced hands, the results of laparoscopic liver surgery are similar to those for laparotomy. PMID- 16273533 TI - Duct-to-mucosa pancreaticogastrostomy is a safe anastomosis following pancreaticoduodenectomy. AB - BACKGROUND: Pancreatic fistula following pancreaticoduodenectomy is relatively common, and remains a major cause of morbidity and mortality. The aim of this study was to evaluate the results of two-layered duct-to-mucosa pancreaticogastrostomy as a method for restoring pancreaticoenteric continuity. METHODS: Prospectively collected data from 100 consecutive patients who underwent Whipple's pancreaticoduodenectomy for tumour were evaluated. All operations were performed by the same surgeon. RESULTS: The perioperative 60-day mortality rate was 1.0 per cent. There were no pancreatic fistulas or anastomotic leaks. Sixteen patients had significant complications that delayed discharge from hospital. Twenty-one patients subsequently required empirical pancreatic exocrine supplements. CONCLUSION: Two-layered duct-to-mucosa pancreaticogastrostomy for restoration of pancreaticoenteric continuity after pancreaticoduodenectomy is associated with a low incidence of complications. PMID- 16273532 TI - Meta-analysis of colonic reservoirs versus straight coloanal anastomosis after anterior resection. AB - BACKGROUND AND METHODS: The comparative benefits and drawbacks of straight coloanal anastomosis (CAA), colonic J-pouch and coloplasty anastomosis after anterior resection are uncertain. Studies published between 1986 and 2005 of colonic J-pouch versus transverse coloplasty or straight CAA were analysed. Endpoints included postoperative complications, and functional and physiological outcomes measured within 6 months, 1 year and 2 years or more after the procedure. A random-effect model was used to aggregate the study endpoints and assess heterogeneity. RESULTS: Thirty-five studies containing 2240 patients (1066 straight CAA, 1050 J-pouch and 124 coloplasty) were included. There was no significant difference in postoperative complications between the three groups. There was a significant reduction in the frequency of defaecation per day by 1.88, 1.35 and 0.74 motions at the three time intervals in the J-pouch group compared with the straight CAA group. Faecal urgency was less prevalent in patients with a J-pouch than those with a straight CAA (odds ratio 0.27 at 6 months or less and 0.21 at 1 year). There was no difference in functional outcome between J-pouch and coloplasty anastomosis. CONCLUSIONS: The colonic J-pouch provided functional benefits over straight anastomosis with no increase in postoperative complications. Coloplasty appeared to have similar benefits but further studies are required for validation. PMID- 16273535 TI - Recent progress towards a molecular understanding of Marfan syndrome. AB - Marfan syndrome (MFS) is a systemic disorder of the connective tissue that is inherited as an autosomal dominant trait and which displays variable manifestations in the ocular, skeletal, and cardiovascular systems. These pleiotropic manifestations are accounted for by mutations in fibrillin-1, the building block of extracellular microfibrils. During the past 10 years, we have witnessed significant progress in delineating the pathological events responsible for the manifestations of MFS. Much of this progress has been based on the creation and analysis of fibrillin-1 mutant mouse lines that faithfully recapitulate the spectrum of clinical severity of MFS. These studies have established the critical contribution of fibrillin-1 deficiency to disease progression through altered cell-matrix interactions and dysregulated TGF-beta signaling. As a result, our definition of MFS as the prototypical structural disorder of the connective tissue has changed to that of a developmental abnormality with broad and complex effects on the morphogenesis and function of multiple organ systems. Importantly, new biological targets have emerged that may yield exciting new opportunities for the development of productive treatment strategies in MFS. PMID- 16273536 TI - Genetic basis of thoracic aortic aneurysms and aortic dissections. AB - Ascending thoracic aortic aneurysms leading to type A dissections (TAAD) can occur in association with a genetic syndrome, such as Marfan syndrome (MFS), or as an autosomal dominant disorder in the absence of syndromic features, termed familial TAAD. Familial TAAD demonstrates genetic heterogeneity, and linkage studies have identified three TAAD loci at 5q13-14 (TAAD1), 11q23 (FAA1), and 3p24-25 (TAAD2). The underlying genetic heterogeneity of TAAD is reflected in the phenotypic variation associated with familial TAAD with respect to age of onset, progression, penetrance, and association with additional cardiac and vascular features. Recently, mutations in the TGFBR2 gene have been identified as the cause of disease linked to the TAAD2 locus, supporting the hypothesis that dysregulation of TGFbeta signaling is a mechanism leading to aneurysms and dissections. The recent identification of the TGFbeta pathway as a key target in the molecular pathogenesis of TAAD has opened new avenues for future genetic and therapeutic research. PMID- 16273537 TI - Differential expression of gamma-aminobutyric acid--a receptor subunits in rat dorsal and ventral hippocampus. AB - Recent data demonstrate weaker gamma-aminobutyric acid (GABA)-ergic inhibition in ventral (VH) compared with dorsal (DH) hippocampus. Therefore, we examined possible differences regarding the GABAA receptors between VH and DH as follows: 1) the expression of the GABAA receptor subunits (alpha1/2/4/5, beta1/2/3, gamma2, delta) mRNA and protein and 2) the quantitative distribution and kinetic parameters of [3H] muscimol (GABAA receptor agonist) binding. VH compared with DH showed: 1) lower levels for alpha1, beta2, gamma2 but higher levels for alpha2 and beta1 subunits in CA1, CA2, and CA3, the differences being more pronounced in CA1 region; in the CA1 region, the mRNA levels of alpha5 were higher, whereas those of alpha4 subunit were slightly lower; in dentate gyrus, the mRNA levels of alpha4, beta3, and delta subunits were significantly lower, presumably suggesting a lower expression of the alpha4/beta3/delta receptor subtype; and 2) lower levels of [3H]muscimol binding, with the lowest value observed in CA1, apparently resulting from weaker binding affinity, insofar as the KD values were higher in VH, whereas the Bmax values were similar between DH and VH. The differences in the subunit expression and the lower affinity of GABAA receptor binding observed predominantly in the CA1 region of VH suggest that the alpha1/beta2/gamma2 GABAA receptor subtype dominates in DH, and the alpha2/beta1/gamma2 subtype prevails in VH. This could underlie the lower GABAA-mediated inhibition observed in VH and, to some extent, explain 1) the higher liability of VH for epileptic activity and 2) the differential involvement of DH and VH in cognitive and emotional processes. PMID- 16273538 TI - Estrogen differentially regulates expression of Per1 and Per2 genes between central and peripheral clocks and between reproductive and nonreproductive tissues in female rats. AB - Although it has long been established that estrogen alters circadian rhythms in behavior, physiology, and reproductive functions in mammals, the molecular mechanism for these effects remains unknown. To explore the possibility that estrogen affects circadian rhythms by changing the expression of clock-related genes, we investigated the effects of chronic treatment with 17beta-estradiol (E2) on the expression of Per1 and Per2 genes in the brain (suprachiasmatic nucleus and cerebral cortex) and periphery (liver, kidney, and uterus) of ovariectomized rats by means of in situ hybridization and northern blotting. In the brain, E2 treatment advanced the peak of Per2 mRNA expression in the SCN; however, it failed to affect the rhythm of Per2 mRNA expression in the CX and Per1 mRNA expression in both the SCN and the CX. In nonreproductive peripheral tissues (liver and kidney), E2 delayed the phase and increased the amplitude of Per1 mRNA expression. In the reproductive tissues (uterus), biphasic rhythms in Per1 and Per2 mRNA were observed after E2 treatment. These findings suggest that the effects of estrogen are different between central and peripheral clock in the brain, and between reproductive and nonreproductive tissues in the periphery. PMID- 16273539 TI - Factors contributing to neuronal degeneration in retinas of experimental glaucomatous rats. AB - After our studies on ganglion cell degeneration in the glaucomatous retina, the current work further confirmed the reduction of amacrine cells in the retina after the onset of glaucoma. Present study also tried to understand the possible mechanisms underlying neuronal degeneration in the glaucomatous retina. Changes of expressions in immediate early genes (IEGs), glutamate receptors (GluRs), calcium-binding proteins (CaBPs), 8-hydroxy-deoxyguanosine (8-OH-dG) and nitric oxide synthase (NOS), as well as apoptotic-related factors including caspase 3, bax, and bcl-2 were examined. IEGs such as c-fos and c-jun were induced in the retina of the glaucomatous rat as early as 2 hr after the onset of glaucoma and lasted up to 2 weeks. Expressions of GluRs and CaBPs (i.e., parvalbumin and calbindin D-28k) were observed to be increased in the retinal ganglion cell layer (GCL) and inner nuclear layer (INL) at 3 days and 1 week after the onset of glaucoma. The increase occurred well before and during the phase where significant neuronal death was observed in the GCL and INL of the glaucomatous retinae. Induction of 8-OH-dG was present in both the GCL and INL of the glaucomatous retina at 3 days after the onset of glaucoma before significant neuronal death was observed. Furthermore, confocal microscopy study showed the complete colocalization of immunohistochemical expression of caspase 3 with glial fibrillary acidic protein (GFAP), but not with neuronal nuclei (NeuN). It indicates that astrocytes and Muller cells are involved in the pathological processes of neuronal death. The relationship between the linked factors and neuronal degeneration is also discussed. PMID- 16273540 TI - Neural precursor cell susceptibility to human cytomegalovirus diverges along glial or neuronal differentiation pathways. AB - Cytomegalovirus (CMV) is a major cause of congenital brain disease, and its neuropathogenesis may be related to viral infection of rapidly dividing, susceptible neural precursor cells (NPCs). In the present study, we evaluated the susceptibility of human fetal brain-derived NPCs (nestin(+), A2B5(+), CD133(+)) to infection with CMV. Data derived from these studies demonstrated that undifferentiated NPCs supported productive viral replication. After differentiation in the presence of serum, a treatment that promotes development of an astroglial cell phenotype (GFAP(+), nestin(-), A2B5(-)), viral expression was retained. However, differentiation of NPCs in medium containing platelet derived growth factor and brain-derived neurotropic factor, conditions that support the development of neurons (Tuj-1(+), nestin(-), A2B5(-)), resulted in reduced viral expression, with corresponding decreased CMV major immediate-early promoter (MIEP) activity relative to undifferentiated cells. Further experiments showed that cellular differentiation into a neuronal phenotype was associated with elevated levels of various CCAAT/enhancer binding protein beta (C/EBP)-beta isoforms, which suppressed MIEP activity in cotransfected NPCs. Taken together, these data demonstrate that the susceptibility of primary human NPCs to CMV is retained concomitantly with differentiation into glial cells but is actively repressed following differentiation into neurons. PMID- 16273541 TI - Nuclear factor-kappab activation is associated with glutamate-evoked tissue transglutaminase up-regulation in primary astrocyte cultures. AB - We have previously demonstrated that alterations of cell redox state, evoked by glutamate, are associated with tissue transglutaminase increases in primary astrocyte cultures. Furthermore, glutamate exposure activated the nuclear factor (NF)-kappaB pathway, and its effects were significantly reduced by antioxidants. Here, we investigated the possible involvement of activated NF-kappaB pathway in glutamate-evoked tissue transglutaminase up-regulation in primary astrocytes. The presence of DNA binding activity by NF-kappaB in nuclear extracts of astrocytes, treated for 24 hr with glutamate (500 microM) or untreated, was assessed by EMSA, using an oligonucleotide probe containing the NF-kappaB consensus sequence present in the tissue transglutaminase promoter. Supershifting with monoclonal antibodies revealed that activated NF-kappaB dimer complexes were composed of p50 and p65 subunits. Interestingly, the specific NF-kappaB inhibitor SN50 (but not its inactive analogue SN50M), when added to cell cultures 30 min prior to glutamate treatment, was able gradually to reduce glutamate-induced NF-kappaB activation. Western blot analysis confirmed the reduction of the p50 amount in nuclear extracts. Notably, the preincubation with SN50 also diminished glutamate increased tissue transglutaminase expression, as showed by both RT-PCR and Western blotting. Competition experiments, carried out with an excess of a probe containing the NF-kappaB consensus sequence present in the kappa-light-chain promoter, demonstrated a preferential binding of the tissue transglutaminase specific NF-kappaB probe in the nuclear extracts of glutamate-treated astrocytes compared with untreated astrocytes. These preliminary data suggest that NF-kappaB activation, which has been demonstrated to be involved in astrocyte response to glutamate, could also be associated with the molecular pathway leading to glutamate-evoked tissue transglutaminase up-regulation. PMID- 16273542 TI - Allopregnanolone treatment, both as a single injection or repetitively, delays demyelination and enhances survival of Niemann-Pick C mice. AB - Niemann-Pick C disease (NPC) is an irreversible neurodegenerative disorder without current treatment. It is thought to result from deficient intracellular cholesterol and/or ganglioside trafficking. We have investigated the effects of allopregnanolone treatments on survival, weight loss, motor function, magnetic resonance imaging (MRI), and neuropathology in the mouse model of NPC (Npc1(-/-) mice). We confirmed previous results showing that a single injection of 250 microg of allopregnanolone on postnatal day 7 significantly extended the life span of Npc1(-/-) mice. This caused a marked difference in the weight curves of the treated mice but no statistical difference in the Rota-Rod performance. T2 weighted MRI and diffusion tensor imaging (DTI) of treated mice showed values of signal intensity and fractional anisotropy closer to those of wild-type mice than those of untreated Npc1(-/-) mice. Neuropathology showed that day-7 treatment markedly suppressed astrocyte reaction and significantly reduced microglial activation. Furthermore, the steroid treatment also increased myelination in brains of Npc1(-/-) mice. Similar effects of allopregnanolone treatment were observed in Npc1(-/-), mdr1a(-/-) double-mutant mice, which have a deficient blood-brain barrier, resulting in increased steroid uptake. The effects on survival and weight loss of a single injection on day 7 followed by injections every 2 weeks were also evaluated in Npc1(-/-) mice, and the beneficial effects were found to be greater than with the single injection at day 7. We conclude that allopregnanolone treatment significantly ameliorates several symptoms of NPC in Npc1(-/-) mice, presumably by effects on myelination or neuronal connectivity. PMID- 16273543 TI - Activation and circuitry of uterine-cervix-related neurons in the lumbosacral dorsal root ganglia and spinal cord at parturition. AB - Stimulation of the uterine cervix at parturition activates neural circuits involving primary sensory nerves and supraspinally projecting neurons of the lumbosacral spinal cord, resulting in output of hypothalamic neurohormones. Dorsal root ganglia (DRG) and spinal neurons of these circuits are not well characterized. The objectives of this study were to detail the activation of DRG and spinal neurons of the L6/S1 levels that are stimulated at late pregnancy, verify hypothalamic projections of activated spinal neurons, and determine whether activated neurons express estrogen receptor-alpha (ERalpha). Expression of phosphorylated cyclic-AMP response element-binding protein (PCREB) and Fos immunohistochemistry were used to "mark" activated DRG and spinal neurons, respectively. Retrograde tracing identified uterine-cervix-related and spinohypothalamic neurons. Baseline PCREB expression in the DRG increased during pregnancy and peaked during the last trimester. Some PCREB-expressing neurons contained retrograde tracer identifying them as cervix-related neurons. Fos expressing neurons were few in spinal cords of nonpregnant and day 22 pregnant rats but were numerous in parturient animals. Some Fos-expressing neurons located in the dorsal half of the spinal cord contained retrograde tracer identifying them as spinohypothalamic neurons. Some DRG neurons expressing PCREB also expressed ERalpha, and some spinal neurons activated at parturition projected axons to the hypothalamus and expressed ERalpha. These results indicate that DRG and spinal cord neurons are activated at parturition; that those in the spinal cord are present in areas involved in autonomic and sensory processing; that some spinal neurons project axons to the hypothalamus, ostensibly part of a neuroendocrine reflex; and that sensory and spinal neurons can respond to estrogens. Moreover, some activated sensory neurons may be involved in the animal's perception of labor pain. PMID- 16273544 TI - Molecular alterations resulting from frameshift mutations in peripheral myelin protein 22: implications for neuropathy severity. AB - Alterations in peripheral myelin protein 22 (PMP22) expression are associated with a heterogeneous group of hereditary demyelinating peripheral neuropathies. Two mutations at glycine 94, a single guanine insertion or deletion in PMP22, result in different reading frameshifts and, consequently, an extended G94fsX222 or a truncated G94fsX110 protein, respectively. Both of these autosomal dominant mutations alter the second half of PMP22 and yet are linked to clinical phenotypes with distinct severities. The G94fsX222 is associated with hereditary neuropathy with liability to pressure palsies, whereas G94fsX110 causes severe neuropathy diagnosed as Dejerine-Sottas disease or Charcot-Marie-Tooth disease type IA. To investigate the subcellular changes associated with the G94 frameshift mutations, we expressed epitope-tagged forms in primary rat Schwann cells. Biochemical and immunolabeling studies indicate that, unlike the wild-type protein, which is targeted for the plasma membrane, frameshift PMP22s are retained in the cell, prior to reaching the medial Golgi compartment. Similar to Wt-PMP22, both frameshift mutants are targeted for proteasomal degradation and accumulate in detergent-insoluble, ubiquitin-containing aggregates upon inhibition of this pathway. The extended frameshift PMP22 shows the ability to form spontaneous aggregates in the absence of proteasome inhibition. On the other hand, Schwann cells expressing the truncated protein proliferate at a significantly higher rate than Schwann cells expressing the wild-type or the extended PMP22. In summary, these results suggest that a greater potential for PMP22 aggregation is associated with a less severe phenotype, whereas dysregulation of Schwann cell proliferation is linked to severe neuropathy. PMID- 16273545 TI - Antioxidant and antiinflammatory activities of xanthorrhizol in hippocampal neurons and primary cultured microglia. AB - Xanthorrhizol, a natural sesquiterpenoid isolated from the rhizome of Curcuma xanthorrhiza Roxb (Zingiberaceae), has antibacterial activities and protective effects against cisplatin-induced hepatotoxicity. In this study, we investigated the activities of xanthorrhizol as an antioxidant or antiinflammatory agent using neuronal and microglial cells. Xanthorrhizol had potent neuroprotective effects on glutamate-induced neurotoxicity and reactive oxygen species (ROS) generation in the murine hippocampal HT22 cell line. Also, xanthorrhizol inhibited H(2)O(2) induced lipid peroxidation in rat brain homogenates. The properties of xanthorrhizol as an antiinflammatory agent were investigated in microglial activation by lipopolysaccharide. It reduced the expression of cyclooxygenase-2 and the inducible nitric oxide synthase, which consequently resulted in the reduction of nitric oxide. The production of proinflammatory cytokines, such as interleukin-6 and tumor necrosis factor-alpha in activated microglial cells, was reduced by xanthorrhizol. These results suggest that xanthorrhizol could be an effective candidate for the treatment of Alzheimer's disease- and other neurological disease-related ROS and inflammation. PMID- 16273547 TI - Immunolocalization of NFATc4 in the adult mouse brain. AB - NFATc4 has recently been identified as playing an important role in variety of activity-dependent neuronal processes, including hippocampal plasticity, axonal growth, neuronal survival, and apoptosis. However, a systematic study examining the distribution of NFATc4 within the nervous system has yet to be conducted. With this in mind, we sought to determine the regional localization of NFATc4 within the adult mouse brain. Interestingly, NFATc4 was expressed broadly, but not uniformly, throughout various brain structures. The highest levels of NFATc4 expression were localized to the hippocampus, olfactory bulb, and various hypothalamic nuclei. Other brain regions that expressed NFATc4 included the cerebellum, striatum, globus pallidus, amygdala, neocortex, and brainstem nuclei. Given NFATc4's widespread expression, these results are consistent with the notion that NFATc4 may underlie activity-dependent neuronal plasticity throughout the adult brain. PMID- 16273548 TI - Sleep deprivation-induced protein changes in basal forebrain: implications for synaptic plasticity. AB - The need to sleep is universal and lack of sleep often results in decreases in alertness and cognitive function. Data suggest that sleep-related mechanisms and deficits resulting from loss of sleep are associated anatomically with the basal forebrain. Long-term effects of sleep deprivation, those lasting a day or more, likely require transcriptional changes leading to changes in protein synthesis, whereas short-term effects may be mediated by rapid changes in the functional status of proteins. To understand sleep deprivation-induced changes in proteins in the wake-promoting area of the basal forebrain in rat, proteomic analysis was carried out by a combination of 2D gel electrophoresis to separate and visualize proteins and matrix-assisted laser desorption/ionization time-of flight mass spectrometry for protein identification. Among 969 protein spots that were compared, 89 spots showed more than a twofold difference between 6-hr sleep deprived rats and undisturbed sleeping controls. We have identified 11 of these proteins to be either cytoskeletal or associated with synapses. The changes in four of these proteins were analyzed further by Western blots of 1D and 2D. Two proteins associated with the cytoskeleton, tubulin and GAP43, show posttranslational modifications. Increased tyrosination of alpha tubulin isoforms and increased phosphorylation of GAP43 was observed after 6-hr sleep deprivation when compared to that in sleeping controls. The synaptic protein synaptosomal associated protein-25 (SNAP25) is decreased whereas amphiphysin is phosphorylated after sleep deprivation. These changes in proteins in the basal forebrain during short-term sleep deprivation are suggestive of changes in the substrate for neuronal transmission and plasticity. PMID- 16273546 TI - Modulation of excitatory synaptic transmission by endogenous glutamate acting on presynaptic group II mGluRs in rat substantia nigra compacta. AB - Excitatory synaptic inputs from the subthalamic nucleus (STN) have been proposed to underlie burst firing of substantia nigra pars compacta (SNc) dopamine (DA) neurons in Parkinson's disease. Given the potential importance of the STN-SNc synapse in health and disease, our goal was to study how transmission at this synapse is regulated. We tested the hypothesis that neurotransmission at STN-SNc synapses is tonically inhibited by endogenous glutamate acting on presynaptic group II metabotropic glutamate receptors (mGluRs). By using whole-cell recording techniques in brain slices, we examined the effect of LY341495, a mGluR antagonist that is most potent at group II mGluRs, on excitatory postsynaptic currents (EPSCs) that either were evoked in SNc DA neurons by stimulation of the STN or were spontaneously occurred in the presence of tetrodotoxin (miniature EPSCs; mEPSCs). LY341495 increased the evoked EPSC amplitude and mEPSC frequency without changing mEPSC amplitude. In contrast, the group III mGluR antagonist UBP1112 failed to increase the evoked EPSC amplitude. An elevation of extracellular glutamate concentration by a glutamate transporter inhibitor, TBOA, suppressed the evoked EPSCs. LY341495, but not UBP1112, partially reversed the TBOA action. The modulations of EPSCs by TBOA and LY341495 were associated with changes in paired-pulse facilitation ratio. Furthermore, TBOA decreased mEPSC frequency, which was partially reversed by LY341495, without affecting mEPSC amplitude. The results indicate that presynaptic group II mGluRs at STN-SNc synapses appear to be partially activated by a basal level of extracellular glutamate and able to sense the change in extracellular glutamate concentration, subsequently modulating synaptic glutamate release. PMID- 16273549 TI - Regeneration of granule neurons after lesioning of hippocampal dentate gyrus: evaluation using adult mice treated with trimethyltin chloride as a model. AB - The hippocampal dentate gyrus in adult animals is known to contain neural progenitors that proliferate and differentiate into neurons in response to brain injury. Little has been observed, however, on regeneration of the granule cell layer of the dentate gyrus that has been directly injured. Using trimethyltin (TMT)-treated mice as an in vivo model, we evaluated the ability of this layer to regenerate after injury. The administration of TMT induced neuronal death in the dentate gyrus selectively 2 days later, with recovery of granule neurons on day 14 and thereafter. At an early stage (days 2-5) after the damage by TMT treatment, 5-bromo-2'-deoxyuridine (BrdU) incorporation into at least two different types of cells was facilitated in the dentate gyrus: BrdU positive/neuronal nuclear antigen (NeuN)-negative cells were found predominantly in the subgranular zone and granule cell layer, whereas BrdU-positive/NeuN positive cells were numerous in the dentate molecular layer and hilus. In addition, expression of proliferating cell nuclear antigen, nestin, NeuroD3, and doublecortin, which are markers for proliferating cells and neural progenitors/neuronal precursors, was extremely enhanced in the dentate gyrus at the early stage after treatment. Double staining revealed that BrdU was colocalized with nestin and doublecortin in the subgranular zone. Behavioral analysis revealed that TMT-induced cognition impairment was ameliorated by day 14 after the treatment. Taken together, our data indicate that the hippocampal dentate gyrus itself is capable of regenerating the neuronal cell layer through rapid enhancement of neurogenesis after injury. PMID- 16273550 TI - Cultured astrocytes from heme oxygenase-1 knockout mice are more vulnerable to heme-mediated oxidative injury. AB - Hemin, the oxidized form of heme, is released from hemoglobin after CNS hemorrhage and may contribute to injury to surrounding tissue. The heme oxygenase (HO) enzymes catalyze the breakdown of hemin to biliverdin, carbon monoxide, and ferric iron. Although HO-2, the isoform expressed predominantly in neurons, accelerates heme-mediated neuronal injury, inhibitor studies suggest that HO-1 induction has a protective effect on astrocytes. In the present study, we directly compared the vulnerability of cultured HO-1 knockout and wild-type astrocytes to hemin. Consistent with prior observations, exposure of wild-type cultures to hemin for 24 hr resulted in protein carbonylation and concentration dependent cell death between 10 and 60 microM, as determined by MTT and lactate dehydrogenase release assays. In cultures prepared from mice lacking the HO-1 gene, oxidative cell injury was approximately doubled. Both protein oxidation and cell death in HO-1 knockout astrocytes were significantly reduced by pretreating cultures with an adenovirus encoding the HO-1 gene prior to hemin exposure. HO-2 expression was observed in both knockout and wild-type cultures and was not altered by HO-1 gene deletion. Cell hemin accumulation after 20 hr hemin exposure was 4.7-fold higher in knockout cells. These results support the hypothesis that HO-1 protects astrocytes from heme-mediated oxidative injury. Selectively increasing its expression in astrocytes may be beneficial after hemorrhagic CNS injuries. PMID- 16273551 TI - Blockade of nicotinic acetylcholine receptors suppresses hippocampal long-term potentiation in wild-type but not ApoE4 targeted replacement mice. AB - Both impaired nicotinic neurotransmission and the inheritance of apoE4 are associated with increased risk for Alzheimer disease (AD) as well as other deficiencies in memory-related behavior. Long-term potentiation (LTP), a cellular model of memory, is known to be altered by nicotinic agents. Recent studies also support an emergent role for apoE in LTP. We compared the effects of mecamylamine, a nonspecific antagonist of nicotinic acetylcholine receptors (nAChRs), on basal synaptic transmission and LTP in hippocampal slices from wild type (wt) mice and targeted replacement mice expressing human apoE4 (apoE4-TR). Field excitatory postsynaptic potentials (EPSPs) were recorded in the dentate gyrus (DG) in response to medial perforant path activation, and theta burst stimulation was used to induce LTP. Bath application of mecamylamine (3 microM) did not alter input-output relationships or paired pulse depression in either mouse strain. Under control conditions, apoE4-TR mice showed significantly less LTP than wt mice (17.5% +/- 3.2%, n = 9, vs. 30.1% +/- 3.9%, n = 11, P < 0.02). Mecamylamine reduced LTP in wt mice to a level that was similar to control levels for apoE4-TR mice (15.7% +/- 3.4%, n = 9), whereas apoE4-TR showed no further reduction of LTP (16.6% +/- 3.7%, n = 8) by mecamylamine. Thus mice expressing human apoE4 differ from wt mice both in their capacity for LTP and in the effect on LTP of nicotinic cholinergic blockade. It is possible that nicotinic neurotransmission is already compromised in apoE4-TR mice and, hence, that interference with the integrity of this cholinergic system represents a mechanism by which inheritance of the apoE4 allele contributes to cognitive risk. PMID- 16273552 TI - Mass spectrometry in bioinorganic analytical chemistry. AB - A considerable momentum has recently been gained by in vitro and in vivo studies of interactions of trace elements in biomolecules due to advances in inductively coupled plasma mass spectrometry (ICP MS) used as a detector in chromatography and capillary and planar electrophoresis. The multi-isotopic (including non metals such as S, P, or Se) detection capability, high sensitivity, tolerance to matrix, and large linearity range regardless of the chemical environment of an analyte make ICP MS a valuable complementary technique to electrospray MS and MALDI MS. This review covers different facets of the recent progress in metal speciation in biochemistry, including probing in vitro interactions between metals and biomolecules, detection, determination, and structural characterization of heteroatom-containing molecules in biological tissues, and protein monitoring and quantification via a heteroelement (S, Se, or P) signal. The application areas include environmental chemistry, plant and animal biochemistry, nutrition, and medicine. PMID- 16273553 TI - Modeling product formation in anaerobic mixed culture fermentations. AB - The anaerobic conversion of organic matter to fermentation products is an important biotechnological process. The prediction of the fermentation products is until now a complicated issue for mixed cultures. A modeling approach is presented here as an effort to develop a methodology for modeling fermentative mixed culture systems. To illustrate this methodology, a steady-state metabolic model was developed for prediction of product formation in mixed culture fermentations as a function of the environmental conditions. The model predicts product formation from glucose as a function of the hydrogen partial pressure (P(H2)), reactor pH, and substrate concentration. The model treats the mixed culture as a single virtual microorganism catalyzing the most common fermentative pathways, producing ethanol, acetate, propionate, butyrate, lactate, hydrogen, carbon dioxide, and biomass. The product spectrum is obtained by maximizing the biomass growth yield which is limited by catabolic energy production. The optimization is constrained by mass balances and thermodynamics of the bioreactions involved. Energetic implications of concentration gradients across the cytoplasmic membrane are considered and transport processes are associated with metabolic energy exchange to model the pH effect. Preliminary results confirmed qualitatively the anticipated behavior of the system at variable pH and P(H2) values. A shift from acetate to butyrate as main product when either P(H2) increases and/or pH decreases is predicted as well as ethanol formation at lower pH values. Future work aims at extension of the model and structural validation with experimental data. PMID- 16273554 TI - Prevention of thermal induced aggregation of cytochrome c at isoelectric pH values by polyanions. AB - Differential scanning calorimetry, viscometry, optical and CD spectroscopy were used to characterize the influence of two polyanions, poly(vinylsulfate) (PVS), and poly(4-styrene-sulfonate) (PSS) on thermal transition reversibility of ferricytochrome c at or near isoelectric pH. In these conditions, both PVS and PSS enhance the thermal transition reversibility of cytochrome c by preventing the aggregation of denatured protein molecules. Data indicate that the polyanions are in complex with cytochrome c that is stabilized by synergistic effect of Coulombic and non-Coulombic interactions. PMID- 16273555 TI - Induction of murine leukemia and lymphoma by dominant negative retinoic acid receptor alpha. AB - Acute promyelocytic leukemia (APL) is invariably associated with chromosomal translocation to retinoic acid receptor alpha (RARalpha) locus. In a vast majority of cases, RARalpha translocates to and fuses with the promyelocytic leukemia (PML) gene. It was thought that the fusion protein PML-RARalpha acts as a double dominant negative mutant to inhibit the PML and RARalpha signaling. In an attempt to study the physiological role of retinoic acid in mammary gland development, we created a transgenic model system expressing a dominant negative RARalpha under the regulation of murine mammary tumor viral promoter. We found that the transgene was also targeted to the lymphoid system in addition to mammary gland. Here we showed that dominant negative RARalpha induced acute lymphoblastic leukemia and lymphoma development in the transgenic mice. Retinoic acid blocked tumor development ex vivo through induction of apoptosis. Thus, our results suggested that disruption of RARalpha signaling was the first essential step in the development of APL in vivo. PMID- 16273557 TI - Ba8CoNb6O24: a d0 dielectric oxide host containing ordered d7 cation layers 1.88 nm apart. PMID- 16273556 TI - Recurrent spontaneous coronary artery dissection. AB - Recurrence of spontaneous coronary artery dissection is a rare event with limited documentation of its existence in the world's literature. We present a case of recurrent spontaneous coronary artery dissection treated with coronary artery bypass surgery and discuss the literature regarding this entity. PMID- 16273559 TI - Nucleobases in molecular recognition: molecular adducts of adenine and cytosine with COOH functional groups. PMID- 16273558 TI - 1,4-migration of rhodium and palladium in catalytic organometallic reactions. AB - Mechanisms based on the 1,4-migration of metal centers have been proposed to explain some interesting transition-metal-catalyzed organic transformations. Despite the lack of solid evidence for the real nature of this process, the 1,4 metal migration reaction is proposed to proceed by the reductive elimination of trivalent intermediates for the Rh-involved reactions and of tetravalent intermediates for the Pd-catalyzed processes. These high-valent organometallic species would be formed by the oxidative addition to C-H and C-X bonds. This minireview summarizes the results in this area. PMID- 16273560 TI - Structural preference versus metal within the MB2C2 (M = Mg, Sc, Ca, Y, Ln) phases: the coloring problem revisited by DFT calculations. PMID- 16273561 TI - Solid-phase oligosaccharide synthesis of a small library of N-glycans. AB - Solid-phase oligosaccharide synthesis is based on a hydroxymethylbenzyl benzoate spacer linker which is connected to the Merrifield resin (1 P). Glycosylation was performed with O-glycosyl trichloroacetimidates of glucosamine, mannose, and galactose permitting chain extension (2e, 5e), branching (4b, 7b, 8b), and chain termination (3t, 6t, 9t) with the use of O-benzyl, O-benzoyl, and N dimethylmaleoyl as permanent and O-fluorenylmethoxycarbonyl (Fmoc) and O phenoxyacetyl (PA) as temporary protecting groups. The steps required on solid phase are i) glycosylation under TMSOTf catalysis, ii) selective cleavage of the temporary protecting groups, Fmoc with NEt3 and PA with 0.5 equivalents of NaOMe in CH2Cl2/MeOH, and iii) product cleavage from the resin with 4.0 equivalents of NaOMe in CH2Cl2/MeOH and following O-acetylation for convenient product isolation. Thus a highly successful synthesis of a small library of seventeen N glycan structures was made possible comprising the N-glycan pentasaccharide core structure 53 and two further chain extended hexa- and heptasaccharide N-glycans with a glucosamine or a lactosamine residue, respectively, which is attached to one of the mannose residues of the core structure (56 and 59). PMID- 16273562 TI - Synthesis and structure of novel organocycloborates. AB - A series of alpha,omega-boryl(bromo)alkanes of the general formula R2B-(CH2)n-Br (n = 3, 4, 5, 6) was obtained in high yield following a standard hydroboration protocol. Upon treatment with Mg turnings and formation of the respective Grignard species, all alkanes with n = 4 to 6 underwent an unprecedented boron centered cyclisation reaction with formation of boratacyclopentanes, -hexanes, and -heptanes, respectively. All new compounds were isolated in high yields as colourless, crystalline materials and characterised in solution by multinuclear NMR spectroscopy. Four representative examples were chosen for X-ray diffraction studies, thus providing the first structurally characterised ring systems of that size at a tetraalkyl borate centre. PMID- 16273563 TI - Domino reactions of 1,3-bis-silyl enol ethers with benzopyrylium triflates: efficient synthesis of fluorescent 6H-benzo[c]chromen-6-ones, dibenzo[c,d]chromen 6-ones, and 2,3-dihydro-1H-4,6-dioxachrysen-5-ones. AB - The condensation of 1,3-bis-silyl enol ethers with benzopyrylium triflates, generated in situ by the reaction of chromones with Me3SiOTf, afforded functionalized 2,3-dihydrobenzopyrans; treatment of the latter with NEt3 or BBr3 resulted in a domino retro-Michael-aldol-lactonization reaction and the formation of a variety of 7-hydroxy-6H-benzo[c]chromen-6-ones. The hydroxy group was functionalized by using Suzuki cross-coupling reactions. The methodology reported was applied to the synthesis of the natural product autumnariol and a new fluorescence dye, which exhibits promising optical properties. 2,3-Dihydro-1H-4,6 dioxachrysen-5-ones were prepared by condensation of chromones with 1,3-bis-silyl enol ethers containing a remote chloride group, domino retro-Michael-aldol lactonization, and an intramolecular Williamson reaction. PMID- 16273564 TI - Vascular response to sirolimus-eluting stents delivered with a nonaggressive implantation technique: comparison of intravascular ultrasound results from the multicenter, randomized E-SIRIUS, and SIRIUS trials. AB - BACKGROUND: The effectiveness of SES to reduce the risk of restenosis was initially demonstrated in short lesions using stent implantation with routine pre dilatation and post-dilatation. This intravascular ultrasound (IVUS) substudy of the E-SIRIUS trial sought to evaluate local arterial responses to sirolimus eluting stents (SES) delivered with a stent implantation technique allowing direct stenting and only selectively applying high-pressure post-dilatation. METHODS AND RESULTS: IVUS was performed immediately after intervention and at 8 month follow-up in 51 patients randomised to either bare-metal stents (BMS; Bx Velocitytrade mark; N=20) or SES (Cyphertrade mark N=31). Direct stenting was allowed (24%) and post-dilation was performed only selectively (32%). Lumen dimensions, intimal hyperplasia and vessel remodeling were compared between SES and BMS. Subsequently, results of SES in the E-SIRIUS IVUS substudy (N=31) were compared to those of SES in the IVUS substudy of the SIRIUS trial (N=137). SES in SIRIUS IVUS substudy were delivered with 100% pre-dilatation and 77% post dilatation. Baseline stent and reference segment measurements were similar between BMS and SES in E-SIRIUS IVUS patients. Using SES there was a 96% reduction in intimal hyperplasia volume within the stented segment (1.8+/-4.9 vs 50.6+/-39.7 mm3, P<0.001) and a significantly larger minimal lumen cross sectional area at 8-month follow-up (4.5+/-1.1 vs 2.3+/-0.9 mm2, P<0.001). No vessel remodeling was observed with the use of SES. The applied stent implantation technique resulted in a minimal stent/reference vessel area ratio of 0.75+/-0.17 in E-SIRIUS SES as compared to 0.84+/-0.23 in SIRIUS SES (P=0.046). Mean intimal hyperplasia cross-sectional area at follow-up was 0.1+/-0.2 mm2 in the SES group of E-SIRIUS and 0.5+/-0.8 mm2 in the SES group of SIRIUS (P=0.003). CONCLUSIONS: An implantation technique of SES which includes direct stenting and minimizes the use of high-pressure post-dilatation results in less optimal stent expansion. However, follow-up results compare very favourable to those of BMS and are characterised by even less intimal hyperplasia than after a more forceful implantation of SES. PMID- 16273565 TI - Ultrafast intramolecular charge transfer and internal conversion with tetrafluoro aminobenzonitriles. AB - The five 2,3,5,6-tetrafluoro-4-aminobenzonitriles XABN4F with a dimethyl-amino (DMABN4F), diethyl-amino (DEABN4F), azetidinyl (AZABN4F), methyl-amino (MABN4F) or amino (ABN4F) group undergo ultrafast intramolecular charge transfer (ICT) at room temperature, in the polar solvent acetonitrile (MeCN) as well as in the nonpolar n-hexane. ICT also takes place with the corresponding non-fluorinated aminobenzonitriles DMABN, DEABN and AZABN in MeCN, whereas for these molecules in n-hexane only minor (DMABN, DEABN) or no (AZABN) ICT fluorescence is detected. For the secondary (MABN) and primary (ABN) amines, an ICT reaction does not occur, which makes ABN4F the first electron donor/acceptor molecule with an NH(2) group for which ICT is observed. The ICT state of the XABN4Fs has a dipole moment of around 14 D, clearly smaller than that of DMABN (17 D). This difference is attributed to the electron withdrawing from the CN group to the phenyl ring, exerted by the four F-substituents. The reaction from the initially prepared locally excited (LE) to the ICT state in n-hexane proceeds in the sub-picosecond time range: 0.35 ps (DMABN4F), 0.29 ps (DEABN4F) and 0.13 ps (AZABN4F), as determined from femtosecond transient absorption measurements. In the highly polar solvent MeCN, an ICT reaction time of around 90 fs is observed for all five XABN4Fs, irrespective of the nature of their amino group. This shows that with these molecules in MeCN the ICT reaction rate is limited by the solvent dielectric relaxation time of MeCN, for which a value of around 90 fs has been reported. It is therefore concluded that, during this ultrashort ICT reaction, a large-amplitude motion such as a full 90 degrees twist of the amino group is unlikely to occur in the XABN4Fs. The ICT state of the XABN4Fs is strongly quenched via internal conversion (IC), with a lifetime tau'(0) (ICT) down to 3 ps, possibly by a reaction passing through a conical intersection made accessible due to a deformation of the phenyl group by out-of-plane motions induced by vibronic coupling between low-lying pisigma* and pipi* states in the XABN4Fs. PMID- 16273566 TI - Performance of fluorescence correlation spectroscopy for measuring diffusion and concentration. AB - Fluorescence correlation spectroscopy (FCS) has become an important tool for measuring diffusion, concentration, and molecular interactions of cellular components. The interpretation of FCS data critically depends on the measurement set-up. Here, we present a rigorous theory of FCS based on exact wave-optical calculations. Six of the most important optical and photophysical factors that influence FCS are studied: fluorescence anisotropy, cover-slide thickness, refractive index of the sample, laser-beam geometry, optical saturation, and pinhole adjustment. Our theoretical framework represents a general attempt to link all relevant parameters of the experimental set-up with the measured correlation function. PMID- 16273567 TI - Energy-transfer efficiency in stacked oligo(p-phenylene vinylene)s: pronounced effects of order. AB - The supramolecular structure of two types of oligo(p-phenylene vinylene) (OPV) building blocks in dodecane solution is studied. Monofunctional chromophores (MOPV) form well-defined helical assemblies, whereas bifunctional molecules (BOPV) aggregate into so-called frustrated stacks, lacking any higher helical order. This difference in organization has a major influence on the transfer of excitation energy through the assemblies. Energy transfer to supramolecularly incorporated guests (MOPV with lower bandgap) is used to probe the intrinsic differences in exciton mobility in these two types of mixed aggregates. From the observed donor fluorescence quenching, it can be concluded that the helically ordered nature of the MOPV stacks facilitates the transfer of excitation energy, yielding evidence for higher exciton mobility in the well-ordered assemblies than in the frustrated stacks. Finally, the concept of energy transfer in supramolecular assemblies is extended to the solid state by the successful implementation in a light-emitting diode (LED). PMID- 16273568 TI - Photophysical properties of BODIPY-derived hydroxyaryl fluorescent pH probes in solution. AB - The photophysical properties of five fluorescent pH probes derived from 4,4 difluoro-4-bora-3a,4a-diaza-s-indacene with phenolic or naphtholic subunits at position 8 and with substituents having different electron driving forces at positions 3 and 5 have been investigated in several organic solvents, by means of absorption, steady-state, and time-resolved fluorimetry. For each compound, the fluorescence quantum yield and lifetime are lower in solvents with higher polarity, owing to an increase in the rate of nonradiative deactivation. The rate constants for radiative deactivation, k(f), are nearly constant for all dyes in all solvents studied [k(f)=(1.7+/-0.2)x10(8) s(-1)]. In aqueous solution, these probes undergo a reversible protonation-deprotonation in the near-neutral to basic pH range, producing intensity increases with lower pH. The pK(a) values of the indicators are between 7.5 and 9.3, depending on the substitution pattern on positions 3, 5, and 8. The difference between the absorption and excitation spectra as a function of pH is indicative of the presence of two species in aqueous solution: the phenol- or naphthol-based indicator and its conjugate base. PMID- 16273569 TI - Identifiability of models for fluorescence quenching in aqueous micellar systems. AB - The first deterministic identifiability analysis is presented for four commonly used kinetic models of fluorescence quenching of an excited probe in aqueous micelles: A) model with immobile quenchers, B) model with mobile quenchers, C) an extension of model B in which exchange of quenchers both via the aqueous phase and during micelle collisions is taken into account, and D) model with probe migration. It is shown that these specific models for fluorescence decay of an excited probe solubilized in a micelle and quenched by molecules or ions that are Poisson-distributed over the micelles, resulting in the generalized four parameter equation f(t)=A(1) exp{-A(2)t-A(3)[1-A(4)t]}, are uniquely identifiable in terms of four descriptive A parameters. Moreover, each model also can be uniquely identified in terms of the underlying rate constants and micellar concentration or mean micellar aggregation number. This means that these parameters can be extracted in a unique way from time-resolved fluorescence quenching experiments on a probe in micelles. For each model the recommended analysis approach is given. PMID- 16273570 TI - Artificial photosynthetic reaction centers: mimicking sequential electron and triplet-energy transfer. AB - An artificial photosynthetic reaction center consisting of a carotenoid (C), a dimesitylporphyrin (P), and a bis(heptafluoropropyl)porphyrin (P(F)), C-P-P(F) , and the related triad in which the central porphyrin has been metalated to give C P(Zn)-P(F) have been synthesized and characterized by transient spectroscopy. These triads are models for amphipathic triads having a carboxylate group attached to the P(F) moiety; they are designed to carry out redox processes across lipid bilayers. Triad C-P-P(F) undergoes rapid singlet-singlet energy transfer between the porphyrin moieties, so that their excited states are in equilibrium. In benzonitrile, photoinduced electron transfer from the first excited singlet state of P and hole transfer from the first excited singlet state of P(F) yield the initial charge-separated state C-P(.) (+)-P(F) (.) (-). Subsequent hole transfer to the carotenoid moiety generates the final charge separated state C(.) (+)-P-P(F) (.) (-), which has a lifetime of 1.1 mus and is formed with a quantum yield of 0.24. In triad C-P(Zn)-P(F) energy transfer from the P(Zn) excited singlet to the P(F) moiety yields C-P(Zn)-(1)P(F) . A series of electron-transfer reactions analogous to those observed in C-P-P(F) generates C(.) (+)-P(Zn)-P(F) (.) (-), which has a lifetime of 750 ns and is formed with a quantum yield of 0.25. Flash photolysis experiments in liposomes containing an amphipathic version of C-P(Zn)-P(F) demonstrate that the added driving force for photoinduced electron transfer in the metalated triad is useful for promoting electron transfer in the low-dielectric environment of artificial biological membranes. In argon-saturated toluene solutions of C-P-P(F) and C-P(Zn)-P(F) , charge separation is not observed and a considerable yield of triplet species is generated upon excitation of the porphyrin moieties. In both triads triplet energy localized in the P(F) moiety is channeled to the carotenoid chromophore by a triplet energy-transfer relay mechanism. Certain photophysical characteristics of these triads, including the sequential electron transfer and the triplet energy-transfer relay mechanism, are reminiscent of those observed in natural reaction centers of photosynthetic bacteria. PMID- 16273571 TI - Image contrast analysis of STM images of self-assembled dioctadecyl chalcogenides on graphite at the liquid-solid interface. AB - The structures of self-assembled monolayers of dioctadecyl selenide (CH3(CH2)17)2Se and dioctadecyl telluride (CH3(CH2)17)2Te, as well as dioctadecyl ether (CH3(CH2)17)2O and dioctadecyl sulfide (CH3(CH2)17)2S, on graphite at the liquid-solid interface were systematically investigated by scanning tunneling microscopy (STM). Both dioctadecyl selenide and telluride formed monolayer structures in which the tilt angle between the molecular axis of the alkyl chain and the lamellae axis was 90 degrees , while dioctadecyl ether assembled with a tilt angle of 60 degrees . Dioctadecyl sulfide was found to make two different self-assembled structures having tilt angles of 60 and 90 degrees . When selenide was embedded in ether compounds in mixed self-assembled monolayers, the alkyl chains of the selenide became blurred, implying that the alkyl chains in the monolayers were unstable. This is in contrast with the structure of co-adsorbed monolayers of the ether and sulfide compounds, where the images of all alkyl chains had high spatial resolution. For the co-adsorbed monolayers, the image contrast of chalcogen atoms was normalized compared with that of alkyl chains of the ether compound in the same image frame. The normalized image contrast was found to be independent of the measurement conditions involving tip shapes, having the following trend, Te>Se>S>C>O. The difference in the normalized image contrast among chalcogen atoms are discussed based on fundamental parameters like polarizability and atomic radii. PMID- 16273572 TI - Scanning tunneling microscopy and spectroscopy of donor-acceptor-donor triads at the liquid/solid interface. AB - By means of scanning tunneling microscopy (STM), the self-assembly of two organic donor-acceptor-donor triads (donor=oligo(p-phenylene vinylene) (OPV); acceptor=perylene diimide (PDI)) and their mixtures has been investigated at the liquid/solid interface. Both triads differ in the nature of the substituents and, therefore, in the redox properties of the central perylene diimide unit (H or Cl). Thanks to the submolecular resolution, the distinct electronic properties of the units, within a triad and between the two triads, are reflected by the relative STM contrast in the bias-dependent imaging experiments. Moreover, scanning tunneling spectroscopy reveals an inverse rectifying behavior of the OPV and H-substituted PDI units, which is discussed in the framework of quasi resonant tunneling. A striking difference is observed for the Cl-substituted triad. PMID- 16273573 TI - Frans De Schryver: forty years of photochemistry and photophysics. PMID- 16273574 TI - Enhancement of Forster energy transfer within a microspherical cavity. AB - Energy transfer from pyrene to perylene molecules co-doped within a poly(methyl methacrylate) latex microsphere was drastically accelerated relative to free space. Fluorescence spectra of the microspheres showed that the relative emission intensities of pyrene and perylene changed with the sphere diameter. Analyses of emission decay profiles clarified that Forster-type energy-transfer processes were induced and that the transfer rates increased within the microspherical cavity. This enhancement can be ascribed not only to the quantum electrodynamic effects on the pyrene emission rate, but also the cavity effect of increasing the overlapping factor between donor emission and acceptor absorption spectra. PMID- 16273575 TI - Energy transfer by a hopping mechanism in dinuclear Ir(III)/Ru(II) complexes: a molecular wire? AB - The synthesis and electrochemical and photophysical properties of a series of heterodinuclear ruthenium-iridium complexes linked by a modular para-phenylene bridge [Ir-ph(n)-Ru]3+ (Ir=Ir(ppyFF)2bpy, Ru=Ru(bpy)3, ppyFF=2-(2,4 difluorophenyl)pyridine), bpy=2,2'-bipyridine, ph=phenylene, n=2, 3, 4, 5) are reported. The use of a high-energy iridium complex, which can act as an energy donor when coupled to the lower energy ruthenium-based component, allows the investigation of photoinduced energy transfer from the excited iridium-centre to the ruthenium fragment (energy acceptor). The rate constants of the energy transfer processes are determined by time-resolved emission and sub-picosecond transient absorption spectroscopy. Interestingly, there is almost no decrease in transfer efficiency or rates as the length between the two chromophores (number of spacers) is increased. This "molecular wire" behavior indicates the dominance of the incoherent hopping mechanism, allowing a very fast energy transfer over long distances (with n = 5 the metal-to-metal distance is estimated to be 32.5 Angstrom). This is the first case in which such behavior is observed for metal complexes, and could lead to new development in molecular electronics. PMID- 16273576 TI - Rapid progression of common femoral artery stenosis leading to development of ischemic foot ulcer following repeated ipsilateral arterial access. AB - Common femoral artery stenosis is well appreciated in patients with peripheral arterial disease, but not described following repeated arterial access. We present a case of progressive right common femoral artery stenosis resulting in resting leg ischemia and ulceration that was temporally related to multiple ipsilateral vascular access procedures for recurrent coronary and peripheral diagnostic and therapeutic procedures. We hypothesize that recurrent arterial injury in a setting of diabetes resulted in progressive critical common femoral artery stenosis. PMID- 16273578 TI - Ultrafast electron diffraction: dynamical structures on complex energy landscapes. AB - In this contribution, we report studies in ultrafast electron diffraction (UED), with the aim of exploring new directions. The main focus is on the determination of complex structures and their dynamics with spatial and temporal resolutions sufficient to give an atomic-scale picture for the evolution in chemical or biological change. We also provide the theoretical framework for UED, and compare the experimental findings of UED to those predicted by density functional and charge density calculations. Selected applications are given in order to highlight phenomena related to concepts such as bifurcation of trajectories in dynamics, far-from-equilibrium coherent structures, and conformational robustness in biological structures. For the former two cases, we consider chemical systems, and, for the latter, we examine proteins of 200 atoms (angiotensin I) or more. PMID- 16273579 TI - Long-lived charge-transfer states in compact donor-acceptor dyads. AB - Photoinduced electron transfer is a widely applied method to convert photon energy into a useful (electro)chemical potential, both in nature and in artificial devices. There is a continuing effort to develop molecular systems in which the charge-transfer state, populated by photoinduced electron transfer, survives sufficiently long to tap the energy stored in it. In general this has been found to require the construction of rather complex molecular systems, but more recently a few approaches have been reported that allow the use of much more simple and relatively small electron donor-acceptor dyads for this purpose. The most successful examples of such systems seem to be those that apply "electron spin control" to slow down the spontaneous decay of the charge-transfer state, and these are reviewed in this minireview, with a discussion of the underlying principles and a critical evaluation of some of the claims made with regard to using a pronounced "inverted-region effect" as an alternative method to prolong the lifetime of charge-transfer states. PMID- 16273580 TI - Ultrafast electron diffraction: oriented molecular structures in space and time. AB - The technique of ultrafast electron diffraction allows direct measurement of changes which occur in the molecular structures of isolated molecules upon excitation by femtosecond laser pulses. The vectorial nature of the molecule radiation interaction also ensures that the orientation of the transient populations created by the laser excitation is not isotropic. Here, we examine the influence on electron diffraction measurements--on the femtosecond and picosecond timescales--of this induced initial anisotropy and subsequent inertial (collision-free) molecular reorientation, accounting for the geometry and dynamics of a laser-induced reaction (dissociation). The orientations of both the residual ground-state population and the excited- or product-state populations evolve in time, with different characteristic rotational dephasing and recurrence times due to differing moments of inertia. This purely orientational evolution imposes a corresponding evolution on the electron scattering pattern, which we show may be similar to evolution due to intrinsic structural changes in the molecule, and thus potentially subject to misinterpretation. The contribution of each internuclear separation is shown to depend on its orientation in the molecular frame relative to the transition dipole for the photoexcitation; thus not only bond lengths, but also bond angles leave a characteristic imprint on the diffraction. Of particular note is the fact that the influence of anisotropy persists at all times, producing distinct differences between the asymptotic "static" diffraction image and the predictions of isotropic diffraction theory. PMID- 16273582 TI - Eosinophilic tissue response several weeks after sirolimus-eluting Cypher stent implantation within a bare metal stent in a coronary saphenous vein graft. PMID- 16273581 TI - Endovascular stent-graft placement for acute and contained rupture of the descending thoracic aorta. AB - OBJECTIVES: To identify determinants of postinterventional death after endovascular stent-graft placement for acute rupture of the descending thoracic aorta, an emerging therapeutic modality for this highly life-threatening condition. METHODS: Between July 1999 and November 2004, 17 patients (14 males; mean age, 65+/-16 (25-83) years) underwent stent-graft repair of the descending thoracic aorta for acute rupture from a thoracic aneurysm (TAA, n=6), acute aortic dissection (AAD, n=6), penetrating aortic ulcer (PAU, n=3), or blunt chest trauma (n=2). Immediate, 30-day, 1-year, and 3-year mortality was assessed. Twenty-one clinical and procedural variables were evaluated in a post-hoc analysis regarding their influence on mortality. Of these, four preprocedural factors with the greatest impact were used to construct a rupture score with a scale from 0 (no adverse prognostic factors present) to 4 (all four adverse factors present). RESULTS: Stent-graft placement was technically feasible in all patients. Complete exclusion of the ruptured aortic pathology could be achieved in only 11 (65%) patients, despite implantation of 1.6+/-0.9 stent-grafts per patient, with a median length of 130 mm. There was one procedure-related early complication (bleeding at the access site). One patient died immediately following the procedure because of progressive mediastinal hematoma, although the rupture site was effectively sealed. Overall survival rates were (76.5+/-10.3)% at 30 days and (52.9+/-12.1)% at 1 year and remained at (52.9+/-12.1)% at 3 years. The four most important preprocedural denominators of death were (1) TAA or AAD as the underlying etiology of aortic rupture (P=0.024), (2) maximum aortic diameter>5 cm (P=0.024), (3) presence of mediastinal hematoma (P=0.056), and (4) an estimated lesion length requiring >1 stent-graft to be covered (P=0.009). Furthermore, residual leakage at the conclusion of the procedure (P=0.009), postprocedural need for dialysis (P=0.004), and prolonged ventilation (P=0.043) were significantly associated with postprocedural death. Using a threshold of >or=3, the rupture score constructed on the basis of the four preprocedural denominators of death was found to be well suited to discriminate postprocedural death (1-year survival: (20.0+/-12.7)% in patients with a rupture score>or=3 vs. 100% in patients with a rupture score<3, P=0.001). CONCLUSION: Endovascular stent graft placement in patients with acute aortic rupture was technically feasible, albeit still associated with high mortality. A simple risk score constructed in retrospect, on the basis of preprocedural prognostic factors, appeared to provide a useful separation of candidates who are likely to benefit from a straightforward endovascular procedure and should be tested prospectively in future studies. PMID- 16273583 TI - Temperature-dependence of preconditioning for lengthened capillary DNA sequencing. AB - A previous study shows that electrophoretic preconditioning of a commercial polymer solution increases the spacing and resolution of DNA fragments fractionated in this solution by CE at 50 degrees C (Griess, G. A. et al., Electrophoresis 2005, 26, 102). The present study shows that this preconditioning effect on peak spacing progressively increases when the temperature of preconditioning increases to 70 degrees C, though fractionation is still performed at 50 degrees C. An increase in peak sharpness accompanies the increase in peak separation for DNA fragments longer than 200 bases. Changing the preconditioning temperature from 50 to 70 degrees C optimally improves resolution of fragment analysis in the range of 600-2000 nucleotides. When DNA sequencing is performed with automated base calling and 70 degrees C preconditioning at 319 V/cm (47 cm long capillary, Applied Biosystems 310 apparatus), the range of high quality base calls is increased by 25% to 750; the range of low-quality base calls is increased by about 100% to 1200 in comparison to DNA sequencing without preconditioning. PMID- 16273584 TI - Introduction of an single nucleodite polymorphism-based "Major Y-chromosome haplogroup typing kit" suitable for predicting the geographical origin of male lineages. AB - The European Consortium "High-throughput analysis of single nucleotide polymorphisms for the forensic identification of persons--SNPforID", has performed a selection of candidate Y-chromosome single nucleotide polymorphisms (SNPs) for making inferences on the geographic origin of an unknown sample. From more than 200 SNPs compiled in the phylogenetic tree published by the Y Chromosome Consortium, and looking at the population studies previously published, a package of 29 SNPs has been selected for the identification of major population haplogroups. A "Major Y-chromosome haplogroup typing kit" has been developed, which allows the multiplex amplification of all 29 SNPs in a single reaction. Allele genotyping was performed with a single base extension reaction (minisequencing) detected by CE. The validation of the multiplex was performed in a total of 1126 unrelated males distributed among 12 worldwide populations. The approach takes advantage of the specific geographic distribution of the Y chromosome haplogroups and demonstrates the utility of binary polymorphisms to infer the origin of a male lineage. PMID- 16273586 TI - Enhancement of electroosmotic flow using zwitterionic additives. AB - Zwitterionic additives provide a means of altering the EOF without increasing conductivity. The magnitude of the EOF in a bare silica capillary increased by as much as 69% upon addition of 500 mM of zwitterion to the running buffer. The EOF enhancement increases linearly with the zwitterion concentration. With zwitterionic additives of the form +NH3-(CH2)n-COO-, the magnitude of the EOF increase is directly related to the number of methylene groups, (n), which ranges from n = 1 to 7. The endgroups on the zwitterions also affect the EOF enhancement. The effect of Z1-methyl (+N(CH3)3CH2CH2CH2SO3-) on EOF was not a function of either the buffer cation or pH. The EOF enhancement is a function of the dielectric increment of the additive and the nature of the amine functionality. PMID- 16273585 TI - Electrokinetic-driven microfluidic system in poly(dimethylsiloxane) for mass spectrometry detection integrating sample injection, capillary electrophoresis, and electrospray emitter on-chip. AB - A novel microsystem device in poly(dimethylsiloxane) (PDMS) for MS detection is presented. The microchip integrates sample injection, capillary electrophoretic separation, and electrospray emitter in a single substrate, and all modules are fabricated in the PDMS bulk material. The injection and separation flow is driven electrokinetically and the total amount of external equipment needed consists of a three-channel high-voltage power supply. The instant switching between sample injection and separation is performed through a series of low-cost relays, limiting the separation field strength to a maximum of 270 V/cm. We show that this set-up is sufficient to accomplish electrospray MS analysis and, to a moderate extent, microchip separation of standard peptides. A new method of instant in-channel oxidation makes it possible to overcome the problem of irreversibly bonded PDMS channels that have recovered their hydrophobic properties over time. The fast method turns the channel surfaces hydrophilic and less prone to nonspecific analyte adsorption, yielding better separation efficiencies and higher apparent peptide mobilities. PMID- 16273587 TI - Determination of dissociation constants of compounds with potential cognition enhancing activity by capillary zone electrophoresis. AB - Accurate determination of pK(a) values is important for proper characterization of newly synthesized molecules. In this work we have used CZE for determination of pK(a) values of new compounds prepared from intermediates, 2, 3 and 4-(2 chloro-acetylamino)-phenoxyacetic acids, by substituting chloride for 2-oxo pyrrolidine, 2-oxo-piperidine or 2-oxo-azepane. These substances are expected to have a cognition enhancing activity and free radicals scavenging effect. Measurements were performed in a polyacrylamide-coated fused-silica capillary of 0.075 mm ID using direct UV detection at 254 nm. Three electrolyte systems were used for measurements to eliminate effects of potential interactions between tested compounds and components of the BGE. In the pH range 2.7-5.4, chloride, formate, acetate and phosphate were used as BGE co-ions, and sodium, beta-alanine and epsilon-aminocaproate as counterions. Mobility standards were measured simultaneously with the tested compounds for calculations of correct electrophoretic mobilities. Several approaches for the calculation of the pK(a) values were used. The values of pK(a) were determined by standard point-to-point calculation using Henderson-Hasselbach equation. Mobility and pH data were also evaluated by using nonlinear regression. Three parameter sigmoidal function fitted the experimental data with correlation coefficients higher than 0.99. Results from CZE measurements were compared with spectrophotometric measurements performed in sodium formate buffer solutions and evaluated at wavelength where the highest absorbance difference for varying pH was recorded. The experimental pK(a) values were compared with corresponding values calculated by the SPARC online calculator. Results of all three used methods were in good correlation. PMID- 16273588 TI - Separation of proteins with a molecular mass difference of 2 kDa utilizing preparative double-inverted gradient polyacrylamide gel electrophoresis under nonreducing conditions: application to the isolation of 24 kDa human growth hormone. AB - A method for separating proteins with a molecular mass difference of 2 kDa using SDS-PAGE under nonreducing conditions is presented. A sample mixture containing several human growth hormone (hGH) isoforms was initially separated on a weak anion-exchange column. Fractions rich in 24 kDa hGH as determined by analytical SDS-PAGE were pooled and further separated by cation-exchange chromatography. The fractions pooled from the cation-exchange chromatography contained two hGH isoforms with a 2 kDa molecular mass difference according to SDS-PAGE analysis, 22 and 24 kDa hGH. The 22 and 24 kDa hGH were separated using continuous-elution preparative double-inverted gradient PAGE (PDG-PAGE) under nonreducing conditions. The preparative electrophoresis gel was composed of three stacked tubular polyacrylamide matrices, a 4% stacking gel, a 13-18% linear gradient gel, and a 15-10% linear inverted gradient gel. Fractions containing purified 24 kDa hGH were pooled and Western blot analysis displayed immunoreactivity to antihGH antibodies. PDG-PAGE provides researchers with an electrophoretic technique to preparatively purify proteins under nonreducing conditions with molecular mass differences of 2 kDa. PMID- 16273589 TI - Fetal transesophageal echocardiography: clinical introduction as a monitoring tool during cardiac intervention in a human fetus. AB - Because of insufficient imaging by maternal transabdominal fetal echocardiography (TAE) in a human fetus with aortic atresia, imperforate atrial septum and progressive cardiac failure, we assessed the feasibility of fetal transesophageal echocardiography (TEE) as a monitoring tool during fetal cardiac intervention at 24 + 6 weeks of gestation. Percutaneous fetoscopic intraesophageal deployment of the ultrasound catheter was achieved and did not result in any maternal or fetal complications. Fetal TEE permitted substantially clearer definition of fetal cardiac anatomy and intracardiac device manipulations than conventional maternal TAE. Despite the employment of various devices, no sufficiently large opening could be achieved within the atrial septum. Although the fetus tolerated the procedure remarkably well and satisfactory fetoplacental flow could be documented at the end of the procedure, the fetus died from progressive cardiac failure 3 days after the intervention. Fetoscopic TEE is feasible in the human fetus and permits substantially clearer definition of fetal cardiac anatomy and intracardiac manipulations than conventional maternal TAE. Based on the observation of spontaneous closure of multiple iatrogenic perforations of the atrial septum, specialized devices are required in order to improve the technical success rate of septoplasty methods and hence the survival odds of these high risk patients. PMID- 16273590 TI - Impact of a lead glass screen on scatter radiation to eyes and hands in interventional cardiologists. AB - The objective of this study was to assess the impact of a transparent lead glass screen (TLGS) on scatter radiation to the eyes and the hands in interventional cardiologists and to compare the results to the recommended annual threshold values of 150 and 500 mSv, respectively. Local radiation doses to the left eye and the ring finger of the left hand of three operators (A, B, C) were assessed by thermoluminiscence dosimeters during 813 coronary angiographies (CAs), including 190 ad hoc percutaneous coronary interventions (PCIs) either with a TLGS placed between patient and operator [615 CAs including 138 ad hoc PCIs; dose area product (DAP) = 84.9 +/- 71.3 Gy x cm(2)], or without (198 CAs including 52 PCIs; DAP = 85.7 +/- 61.5 Gy x cm(2)). To determine the efficacy of the TLGS, average DAP-normalized local doses were calculated. Using a TLGS, operator A, B, and C performed 259 (in 9 months), 211 (in 8 months), and 145 CAs (in 8 months) with TLGS and acquired cumulative eye lens doses of 5.5, 1.5, and 1.0 mSv corresponding to extrapolated annual doses of 7.3, 2.3, and 1.5 mSv. The cumulative finger doses were 9.6, 10.3, and 6.4 mSv, resulting in extrapolated annual doses of 12.8, 15.5, and 9.6 mSv. Compared to 139 (in 5 months), 36 (in 2 months), and 23 CAs (in 2 months) without TLGS, the use of a TLGS reduced the DAP normalized eye dose by a factor of 19 (with TLGS 0.153 vs. without TLGS 2.924 microSv/Gy x cm(2)), whereas only a weak effect on the dose to the hands was observed (with TLGS 0.504 vs. without TLGS 0.578 microSv/Gy x cm(2)). The consequent use of a TLGS efficiently reduces scatter radiation to the operator's eyes in daily practice, but has only minimal effects on the dose to the hands. PMID- 16273591 TI - Radiation shielding really works, when you use it. PMID- 16273592 TI - Delayed abrupt tamponade by isolated left atrial compression following coronary artery perforation during coronary angioplasty. AB - Cardiac tamponade following coronary artery perforation during percutaneous coronary intervention is a rare but potentially life-threatening complication. When associated with Ellis type III coronary perforations, tamponade develops rapidly during the intervention. In contrast, Ellis type I and II coronary perforations, because of their contained nature, are usually managed conservatively and rarely result in tamponade. We report two unusual cases of delayed but abrupt tamponade caused by localized left atrial compression after contained coronary artery perforation following angioplasty. This complication is an extremely rare event. Etiology, diagnostic modalities, and management are discussed. PMID- 16273593 TI - Acute myeloid leukemia with concurrent myeloid sarcoma treated with autologous bone marrow transplantation: two illustrative cases and a literature review. AB - Myeloid sarcoma (MS) is an invasive extramedullary solid tumor composed of immature cells of the myeloid series. It complicates the clinical course of a minority of patients with acute myeloid leukemia (AML). Traditionally its presence has been regarded as an indicator of aggressive disease. Currently, the optimal treatment of AML with concurrent MS remains to be determined. We report two cases of autologous bone marrow transplantation (auto-BMT) for AML with concurrent MS followed by a review of the literature. PMID- 16273594 TI - Routine pre-evacuation ultrasound diagnosis of hydatidiform mole: experience of more than 1000 cases from a regional referral center. AB - OBJECTIVES: To examine the accuracy of sonographic findings of routine ultrasound examinations in patients with a proven histological diagnosis of complete or partial hydatidiform mole referred to a supra-regional referral center, and to examine the relationship of sonographic findings to gestational age across the first and early second trimesters. METHODS: Review of consecutive cases referred to a trophoblastic disease unit from June 2002 to January 2005 with a diagnosis of possible or probable hydatidiform mole in whom results of a pre-evacuation ultrasound examination were documented. Ultrasound detection rates for partial and complete hydatidiform moles were calculated and comparison of detection rates between complete and partial mole, and gestational age groups carried out. RESULTS: 1053 consecutive cases were examined. The median maternal age was 31 (range, 15-54) years and the median gestational age was 10 (range, 5-27) weeks. 859 had a final review diagnosis of partial or complete hydatidiform mole (82%), including 253 (29%) complete moles and 606 (71%) partial moles. Non-molar hydropic miscarriage was diagnosed following histological review in 194 (18%). Overall, 378 (44%) cases with a final diagnosis of complete or partial hydatidiform mole had a pre-evacuation ultrasound diagnosis suggesting hydatidiform mole, including 200 complete moles and 178 partial moles, representing 79% and 29%, respectively, of those with complete (253) or partial (606) moles in the final review diagnosis. The ultrasound detection rate was significantly better for complete versus partial hydatidiform moles (Z = 13.4, P < 0.001). There was a non-significant trend towards improved ultrasound detection rate with increasing gestational age, with an overall detection rate of 35-40% before 14 weeks' gestation compared to around 60% after this gestation. The sensitivity, specificity, positive predictive value and negative predictive value for routine pre-evacuation ultrasound examination for detection of hydatidiform mole of any type were 44%, 74%, 88% and 23%, respectively. CONCLUSIONS: Routine pre-evacuation ultrasound examination identifies less than 50% of hydatidiform moles, the majority sonographically appearing as missed or incomplete miscarriage. Detection rates are, however, higher for complete compared to partial moles, and improve after 14 weeks' gestation. Histopathological examination of products of conception remains the current gold standard for the identification of gestational trophoblastic neoplasia. PMID- 16273595 TI - Cervical assessment at 22 and 27 weeks for the prediction of spontaneous birth before 34 weeks in twin pregnancies: is transvaginal sonography more accurate than digital examination? AB - OBJECTIVES: This study compared the accuracy of ultrasound cervical assessment (cervical length and cervical index) and digital examination (Bishop score and cervical score) in the prediction of spontaneous birth before 34 weeks in twin pregnancies. METHODS: In a prospective multicenter study, digital examination and transvaginal sonography were performed consecutively in twin pregnancies attending for routine sonography at either 22 weeks (175 women) or 27 weeks (153 women). The digital examination took place first, and the Bishop score and cervical score (cervical length minus cervical dilatation) were calculated. Ultrasound measurements were then made of cervical length and funnel length to yield the cervical index (1 + funnel length/cervical length). The association between each variable and delivery before 34 weeks was tested by the Mann-Whitney U-test. The receiver-operating characteristics (ROC) curves of the ultrasound and digital indicators were determined for both gestational age periods, and the areas under the ROC curves compared. The best cut-off values for each indicator were used to determine predictive values for delivery before 34 weeks. RESULTS: The median gestational age at delivery among the women included in the 22-week examination period was 36.0 (range, 21-40) weeks; 10.9% (19) gave birth spontaneously before 34 weeks. The median cervical length was 40 (range, 6-65) mm. All four parameters were predictors of delivery before 34 weeks. The areas under the ROC curves for cervical index, cervical length, Bishop score and cervical score did not differ significantly. The median gestational age at delivery among the women in the 27-week examination period was 36.0 (range, 27 40) weeks; 9.2% (14) gave birth spontaneously before 34 weeks. The median cervical length was 35 (range, 1-57) mm. All parameters except the Bishop score were predictors of delivery before 34 weeks. The likelihood ratio of the positive and negative tests for cervical length < or = 25 mm was 5.4 (range, 3.2-9.0) and 0.3 (range, 0.1-0.7), respectively, compared with 2.3 (range, 1.3-4.2) and 0.6 (range, 0.3-1.1), respectively, for cervical score < or = 1. The area under the curve for the cervical index was significantly larger than that for the Bishop score (P = 0.008) or cervical score (P = 0.02). CONCLUSION: Transvaginal sonography predicted spontaneous delivery before 34 weeks better than digital examination at the 27-week but not the 22-week examination. PMID- 16273596 TI - Molecular basis for the binding polyspecificity of an anti-cholera toxin peptide 3 monoclonal antibody. AB - The onset of autoimmune diseases is proposed to involve binding promiscuity of antibodies (Abs) and T-cells, an often reported yet poorly understood phenomenon. Here, we attempt to approach two questions: first, is binding promiscuity a general feature of monoclonal antibodies (mAbs) and second, what is the molecular basis for polyspecificity? To this end, the anti-cholera toxin peptide 3 (CTP3) mAb TE33 was investigated for polyspecific binding properties. Screening of phage display libraries identified two epitope-unrelated peptides that specifically bound TE33 with affinities similar to or 100-fold higher than the wild-type epitope. Substitutional analyses revealed distinct key residue patterns recognized by the antibody suggesting a unique binding mode for each peptide. A database query with one of the consensus motifs and a subsequent binding study uncovered 45 peptides (derived from heterologous proteins) that bound TE33. To better understand the structural basis of the observed polyspecificity we modeled the new cyclic epitope in complex with TE33. The interactions between this peptide and TE33 suggested by our model are substantially different from the interactions observed in the X-ray structure of the wild-type epitope complex. However, the overall binding conformation of the peptides is similar. Together, our results support the theory of a general polyspecific potential of mAbs. PMID- 16273597 TI - Prenatal prognosis of congenital diaphragmatic hernia using magnetic resonance imaging measurement of fetal lung volume. AB - OBJECTIVES: To investigate the correlation between fetal lung volume (FLV), measured with magnetic resonance imaging (MRI), and postnatal mortality in newborns with prenatally diagnosed isolated congenital diaphragmatic hernia (CDH). METHODS: In a 4-year prospective multicenter study, 77 fetuses with isolated CDH diagnosed between 20 and 33 weeks' gestation underwent fast spin echo T2-weighted lung MRI. These MRI-FLV measurements were compared with a previously published normative curve obtained in 215 fetuses without thoracoabdominal malformations and with normal ultrasound biometric findings. FLV measurements were correlated with postnatal survival. The mean gestational age at MRI was 31.3 weeks. RESULTS: The measured/expected FLV ratio was significantly lower in the newborns with CDH who died compared with those who survived (23.6 +/ 12.2 vs. 36.1 +/- 13.0, P < 0.001). When the ratio was below 25%, there was a significant decrease in postnatal survival (19% vs. 40.3%, P = 0.008). Survival was significantly lower for neonates when one lung could not be seen by fetal MRI compared with those fetuses with two visible lungs on MRI (17.9% vs. 62.1%, P < 0.001). CONCLUSION: In isolated CDH, FLV measurement by MRI is a good predictor of postnatal mortality due to pulmonary hypoplasia. PMID- 16273598 TI - Fetal OK-432 pleurodesis: complete or incomplete? PMID- 16273599 TI - Inhibitory effects of extracellular adenosine triphosphate on growth of esophageal carcinoma cells. AB - AIM: To study the growth inhibitory effects of ATP on TE-13 human squamous esophageal carcinoma cells in vitro. METHODS: MTT assay was used to determine the inhibition of proliferation of ATP or adenosine (ADO) on TE-13 cell line. The morphological changes of TE-13 cells induced by ATP or ADO were observed under fluorescence light microscope by acridine orange (AO)/ethidium bromide (EB) double stained cells. The internucleosomal fragmentation of genomic DNA was detected by agarose gel electrophoresis. The apoptotic rate and cell cycle after treatment with ATP or ADO were determined by flow cytometry. RESULTS: ATP and ADO produced inhibitory effects on TE-13 cells at the concentration between 0.01 and 1.0 mmol/L. The IC(50) of TE-13 cells exposed to ATP or ADO for 48 and 72 h was 0.71 or 1.05, and 0.21 or 0.19 mmol/L, respectively. The distribution of cell cycle phase and proliferation index (PI) value of TE-13 cells changed, when being exposed to ATP or ADO at the concentrations of 0.01, 0.1, and 1 mmol/L for 48 h. ATP and ADO inhibited the cell proliferation by changing the distribution of cell cycle phase via either G(0)/G(1) phase (ATP or ADO, 1 mmol/L) or S phase (ATP, 0.1 mmol/L) arrest. Under light microscope, the tumor cells exposed to 0.3 mmol/L ATP or ADO displayed morphological changes of apoptosis. A ladder-like pattern of DNA fragmentation was obtained from TE-13 cells treated with 0.1-1 mmol/L ATP or ADO in agarose gel electrophoresis. ATP and ADO induced apoptosis of TE-13 cells in a dose-dependent manner at the concentration between 0.03 and 1 mmol/L. The maximum apoptotic rate of TE-13 cells exposed to ATP or ADO for 48 h was 16.63% or 16.9%, respectively. CONCLUSION: ATP and ADO inhibit cell proliferation, arrest cell cycle, and induce apoptosis of TE-13 cell line. PMID- 16273600 TI - Prognostic implication of isolated tumor cells and micrometastases in regional lymph nodes of gastric cancer. AB - AIM: To determine the prognostic significance of isolated tumor cells (ITCs) and lymph node micrometastases in gastric cancer. METHODS: Hematoxylin and eosin stained slides of lymph node dissections of 632 consecutive gastric cancers were reviewed. Cytokeratin immunostaining was performed in 280 node-negative cases and 5 cases indefinite for lymph node metastases. Lymph node metastases were divided into ITCs, micrometastases, or macrometastases, according to the sizes of tumor deposits in largest dimension. ITCs were further classified into four groups according to metastasis pattern. RESULTS: Lymph node metastases were identified by immunostaining in 58 of 280 node-negative cases (20.7%) and were not significantly associated with patient survival (P = 0.3460). After cytokeratin immunostaining, 196 cases were classified as pN1, which consisted of 20 cases with micrometastases detected by immunostaining (pN1mi(i+)), 34 cases with only micrometastases (pN1mi), and 142 cases with pN1 with one or more macrometastases (pN1). Cases with pN1mi and pN1mi(i+) had a significantly better prognosis than the cases with pN1 (P = 0.0037). ITCs were found in 38 of these 58 cases, and could be divided into four groups: 12 cases with only a single cell pattern, 7 cases with multiple individual cells, 5 cases with single small cluster, and 14 cases with multiple small clusters. Among these four groups, cases with ITCs of multiple individual cell pattern showed the worst survival (median survival: 28 mo, P<0.0001). CONCLUSION: Both size and pattern of lymph node metastases can give prognostic information on the survival of gastric cancer patients. PMID- 16273601 TI - Overexpression of decoy receptor 3 in hepatocellular carcinoma and its association with resistance to Fas ligand-mediated apoptosis. AB - AIM: To characterize the expression and genomic amplification of decoy receptor 3 (DcR3) in hepatocellular carcinoma (HCC) and to evaluate the role of DcR3 in apoptosis. METHODS: We examined 48 cases of HCC for DcR3 expression by RT-PCR and DcR3 gene amplification by quantitative genomic PCR. DcR3 protein was detected by immunohistochemistry. Terminal deoxynucleotidyl transferase-mediated dUTP digoxigenin nick and labeling (TUNEL) was used to identify the apoptosis cells in tissues. Primary hepatoma cell culture and MTT test were used to evaluate the protection against FasL- and chemical-induced apoptosis by DcR3 expression. RESULTS: DcR3 mRNA overexpression was detected in 60% HCC (29/48) patients. The occurrence of HCC was not associated with amplification of the gene. One sample base substitution was found in three sites as sequence in Genbank. The expression of DcR3 in HCC was the associatied with the apoptotic index (0.067+/-0.04 vs 0.209+/-0.12, P<0.01), size of mass, stage and infiltration or metastasis (41.2% vs 71.0%, 40% vs 75%, 51.8% vs 84.6%, P<0.05). DcR3 expression could be protect hepatoma cells against apoptosis induced by FasL, but not by chemicals. CONCLUSION: These data suggest that in addition to gene amplification there may be another mechanism underlying DcR3 overexpression. The effect of overexpression of DcR3 on the apoptosis of cancer cells may have direct therapeutic implications for the management of HCC. PMID- 16273603 TI - Anti-hepatoma effect of arsenic trioxide on experimental liver cancer induced by 2-acetamidofluorene in rats. AB - AIM: To study the anti-hepatoma efficiency of arsenic trioxide (As(2)O(3)) in the treatment of experimental rat hepatocellular carcinoma (HCC) induced by 2 acetamidofluorene (2-FAA) and to elucidate the possible mechanisms. METHODS: SD rats (2 mo old) had been fed with 2-FAA for 8 wk to induce HCC, and then they were treated with As(2)O(3) or matrine. On d 29, the rats were killed and the liver was weighed and liver tumors were counted. The histological changes of liver tissue were observed under microscope, and the cellular dynamic parameters were studied by flow cytometry. Immunohistochemistry (two-step method) was used to observe the expression of vascular endothelial growth factor (VEGF) and micro vessel density (MVD) on consecutive sections. The pathological parameters were also analyzed, the levels of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), total bilirubin (TBi), and direct bilirubin (DBi). RESULTS: The number of liver tumors decreased significantly in groups treated with As(2)O(3), especially in medium-dose (1 mg/kg) group (t = 2.80, P<0.01). As(2)O(3) caused HCC cell death via apoptosis; necrosis was seen and apoptosis was common when the dose was 1 mg/kg. Proliferation index decreased sharply in medium-dose (1 mg/kg) group (7.87+/-4.11 vs 24.46+/-6.49, t = 2087, P<0.01), but not in 0.2 mg/kg group. However, S-phase fraction decreased dramatically in both groups, it reached the bottom level only when the dose was 1 mg/kg compared with control (0.40+/-0.13 vs 3.01+/-0.51, t = 2.97, P<0.01), and it was obviously accompanied with accumulation of cells in G(0)/G(1) (G(0)/G(1) restriction). The expressions of VEGF and MVD in medium-dose (1 mg/kg) group were significantly lower than normal saline group (0.63+/-0.74 vs 2.44+/-0.88, P<0.05; 15.75+/-3.99 vs 47.44+/-13.41, t = 2.80, P<0.01). Compared with normal saline group, medium- and low-dose groups As(2)O(3) and matrine lowered the levels of ALT in serum (61.46+/-9.46, 63.75+/-20.40, 61.18+/-13.00 vs 108.98+/-29.86, t = 2.14, P<0.05), but had no effect on the level of serum AST, TBi, and DBi. CONCLUSION: As(2)O(3) had inhibitory effect on growth of experimental HCC in rats induced by 2-FAA, but had no obvious effect on normal hepatic cells. The mechanisms may involve decrease of cell division, accumulation of cells in G(0)/G(1) phase, apoptosis of tumor cells, and inhibitory effect on angiogenesis through blocking VEGF. PMID- 16273602 TI - Expressions of inducible nitric oxide synthase and matrix metalloproteinase-9 and their effects on angiogenesis and progression of hepatocellular carcinoma. AB - AIM: To determine the expressions of inducible nitric oxide synthase (iNOS) and matrix metalloproteinase-9 (MMP-9) in hepatocellular carcinoma (HCC) and to investigate the relationship between iNOS and MMP-9 expression and their effects on angiogenesis and progression of HCC. METHODS: In this study, we examined iNOS, MMP-9, and CD34 expression in specimens surgically removed from 32 HCC patients and 7 normal liver tissues by immunohistochemical staining. Meanwhile, microvessel density (MVD) was determined as a marker of angiogenesis by counting CD34-positive cells. RESULTS: The positive rates of iNOS and MMP-9 expression were 71.88% (23/32) and 78.13% (25/32) in HCC. MMP-9 expression was significantly correlated with tumor size, capsule status, TNM stage, and risk of HCC recurrence (P = 0.032, P = 0.033, P = 0.007, and P = 0.001, respectively). There was also a significant relationship between iNOS expression and capsule status and risk of HCC recurrence (P = 0.049 and P = 0.004, respectively), but no correlation between iNOS expression and tumor size and TNM stage. There was a positive association between MVD and TNM stage and risk of HCC recurrence (P = 0.037 and P = 0.000, respectively). The count of MVD was significantly different in different iNOS and MMP-9 immunoreactivity groups (F = 17.713 and 17.097, P = 0.000 and P = 0.000, respectively). The examination of Spearman's rank correlation coefficient showed that there was a significant positive correlation between MVD and iNOS, MMP-9 immunoreactivity (r = 0.754 and 0.751, P = 0.000 and P=0.000, respectively). There was also a significant association between MMP-9 and iNOS expression in HCC (P = 0.010). CONCLUSION: Nitric oxide (NO) produced by iNOS could modulate MMP-9 production and therefore contribute to tumor cell angiogenesis and invasion and metastasis in HCC. The strong expression of iNOS and MMP-9 in HCC may be helpful in evaluating the recurrence of HCC, predicting poor prognosis. For patients with strong expression of MMP-9 and iNOS, the optimal treatment scheme needs to be selected. PMID- 16273604 TI - Excess body weight, liver steatosis, and early fibrosis progression due to hepatitis C recurrence after liver transplantation. AB - AIM: To investigate how weight gain after OLT affects the speed of fibrosis progression (SFP) during recurrent hepatitis C virus (HCV) infection of the graft. METHODS: Ninety consecutive patients (63 males, median age 53 years; 55 with HCV-related liver disease), transplanted at a single institution, were studied. All were followed for at least 2 years after OLT and had at least one follow-up graft biopsy, performed not earlier than 1 year after the transplant operation. For each biopsy, a single, experienced pathologist gave an estimate of both the staging according to Ishak and the degree of hepatic steatosis. The SFP was quantified in fibrosis units/month (FU/mo). The lipid metabolism status of patients was summarized by the plasma triglycerides/cholesterol (T/C) ratio. Body mass index (BMI) was measured before OLT, and 1 and 2 years after it. RESULTS: In the HCV positive group, the highest SFP was observed in the first post-OLT year. At that time point, a SFP <=0.100 FU/mo was observed more frequently among recipients who had received their graft from a young donor and had a pre transplant BMI value >26.0 kg/m(2). At completion of the first post-transplant year, a BMI value >26.5 kg/m(2) was associated with a T/C ratio <=1. The proportion of patients with SFP >0.100 FU/mo descended in the following order: female recipients with a high T/C ratio, male recipients with high T/C ratio, and recipients of either gender with low T/C ratio. Hepatic steatosis was observed more frequently in recipients who, in the first post-transplant year, had increased their BMI >=1.5 kg/m(2) in comparison to the pre-transplant value. Hepatic steatosis was inversely associated with the staging score. CONCLUSION: Among HCV positive recipients, excess weight gain post-OLT does not represent a factor favoring early liver fibrosis development and might even be protective against it. PMID- 16273605 TI - Upregulation of cathepsin W-expressing T cells is specific for autoimmune atrophic gastritis compared to other types of chronic gastritis. AB - AIM: To investigate a pathophysiological role of cathepsin W (CatW), a putative thiol-dependent cysteine protease, which is specifically expressed in cytotoxic lymphocytes, in different types of chronic inflammation of the gastric mucosa. METHODS: Gastric and duodenal biopsies of patients with Helicobacter pylori (H pylori)-associated active gastritis (Hp, n = 19), chemically induced reactive gastritis (CG, n = 17), autoimmune atrophic gastritis (AIG, n = 20), lymphocytic corpus gastritis (LG, n = 29), celiac disease (CD, n = 10), and corresponding controls (n = 24) were analyzed by immunohistochemistry for the expression of CatW and CD45. Furthermore, immunohistochemical double staining with anti-CD3 and anti-cathepsin was performed for the samples of AIG. RESULTS: Median values of CatW-expressing cells among CD45-positive immune cells were between 2% and 6% for normal gastric mucosa, CG, and LG, whereas the corresponding value was significantly increased for AIG (24.7%, P<0.001) and significantly decreased for HP (0.7%, P<0.05). Double staining with anti-CD3 and anti-CatW antibodies revealed that >90% of CatW-expressing cells in gastric mucosa of AIG were T cells. Duodenal mucosa had significantly more CatW/CD45-positive cells than normal gastric mucosa (median: 17.8% vs 2%, P<0.01). The corresponding proportion of CatW/CD45-postive cells was decreased in CD compared to duodenal mucosa (median: 2.1% vs 17.8%, P<0.05). CONCLUSION: The opposite findings regarding the presence of CatW-positive cells in AIG (increase) and CD (decrease) reflects the different cellular composition of immune cells involved in the pathogenesis of these diseases. PMID- 16273606 TI - Ischemic preconditioning inhibits development of edematous cerulein-induced pancreatitis: involvement of cyclooxygenases and heat shock protein 70. AB - AIM: To determine whether ischemic preconditioning (IP) affects the development of edematous cerulein-induced pancreatitis and to assess the role of cyclooxygenase-1 (COX-1), COX-2, and heat shock protein 70 (HSP 70) in this process. METHODS: In male Wistar rats, IP was performed by clamping of celiac artery (twice for 5 min at 5-min intervals). Thirty minutes after IP or sham operation, acute pancreatitis was induced by cerulein. Activity of COX-1 or COX-2 was inhibited by resveratrol or rofecoxib, respectively (10 mg/kg). RESULTS: IP significantly reduced pancreatic damage in cerulein-induced pancreatitis as demonstrated by the improvement of pancreas histology, reduction in serum lipase and poly-C ribonuclease activity, and serum concentration of pro-inflammatory interleukin (IL)-1beta. Also, IP attenuated the pancreatitis-evoked fall in pancreatic blood flow and pancreatic DNA synthesis. Serum level of anti inflammatory IL-10 was not affected by IP. Cerulein-induced pancreatitis and IP increased the content of HSP 70 in the pancreas. Maximal increase in HSP 70 was observed when IP was combined with cerulein-induced pancreatitis. Inhibition of COXs, especially COX-2, reduced the protective effect of IP in edematous pancreatitis. CONCLUSION: Our results indicate that IP reduces pancreatic damage in cerulein-induced pancreatitis and this effect, at least in part, depends on the activity of COXs and pancreatic production of HSP 70. PMID- 16273607 TI - Role of nuclear receptor CAR in carbon tetrachloride-induced hepatotoxicity. AB - AIM: To investigate the precise roles of CAR in CCl(4)-induced acute hepatotoxicity. METHODS: To prepare an acute liver injury model, CCl(4) was intraperitoneally injected in CAR+/+ and CAR-/- mice. RESULTS: Elevation of serum alanine aminotransferase and extension of centrilobular necrosis were slightly inhibited in CAR-/- mice compared to CAR+/+ mice without PB. Administration of a CAR inducer, PB, revealed that CCl(4)-induced liver toxicity was partially inhibited in CAR-/- mice compared with CAR+/+ mice. On the other hand, androstanol, an inverse agonist ligand, inhibited hepatotoxicity in CAR+/+ but not in CAR-/- mice. Thus, CAR activation caused CCl(4) hepatotoxicity while CAR inhibition resulted in partial protection against CCl(4)-induced hepatotoxicity. There were no differences in the expression of CYP2E1, the main metabolizing enzyme for CCl(4), between CAR+/+ and CAR-/- mice. However, the expression of other CCl(4)-metabolizing enzymes, such as CYP2B10 and 3A11, was induced by PB in CAR+/+ but not in CAR-/- mice. Although the main pathway of CCl(4)-induced acute liver injury is mediated by CYP2E1, CAR modulates its pathway via induction of CYP2B10 and 3A11 in the presence of activator or inhibitor. CONCLUSION: The nuclear receptor CAR modulates CCl(4)-induced liver injury via induction of CCl(4)-metabolizing enzymes in the presence of an activator. Our results suggest that drugs interacting with nuclear receptors such as PB might play critical roles in drug-induced liver injury or drug-drug interaction even though such drugs themselves are not hepatotoxic. PMID- 16273608 TI - Damaging effects of gliadin on three-dimensional cell culture model. AB - AIM: To evaluate the effects of gliadin on the oxidative environment in the "in vivo-like" model of a three-dimensional cell culture system. METHODS: LoVo cell line (intestinal adenocarcinoma) multicellular spheroids were treated with digested gliadin (with albumin used as a control). Spheroid volumes, cell viability and morphology, lactate dehydrogenase (LDH) release, content of reduced glutathione (GSH) and activity of GSH-related enzymes were examined. The data were statistically analyzed using the Student's t-test. was considered statistically significant. RESULTS: Gliadin reduced cell viability (from 20% to 60%) and led to morphological alterations characterized by apoptotic findings and cytoskeletal injuries. LDH activity increased. The content of GSH reduced (-20% vs controls), and activity of GSH-related enzymes was significantly inhibited. CONCLUSION: Gliadin treatment induces an imbalance in the antioxidative mechanism of cells cultured by the three-dimensional technique. This alteration may explain the cell damage directly caused by gliadin and the subsequent morphological abnormalities. PMID- 16273609 TI - Insulin promotes sinusoidal endothelial cell proliferation mediated by upregulation of vascular endothelial growth factor in regenerating rat liver after partial hepatectomy. AB - AIM: To determine whether insulin could promote sinusoidal endothelial cell (SEC) proliferation mediated by upregulation of vascular endothelial growth factor (VEGF) in regenerating rat liver after partial hepatectomy (PHx). METHODS: Adult male Sprague-Dawley rats undergoing 70% PHx were injected with insulin (300 MU/kg) or saline via the tail veins every 8 h after surgery for 7 d and killed at 0, 24, 48, 72, 96, 120, 144, and 168 h after surgery. Proliferation of both hepatocytes and SECs was monitored by evaluating the proliferating cell nuclear antigen (PCNA) labeling index (LI). The expression of VEGF protein was evaluated by immunohistochemistry. The mRNA expressions of VEGF and its receptors Flt-1 and Flk-1 were evaluated by semi-quantitative reverse transcription-PCR. RESULTS: Insulin markedly increased the expression of VEGF mRNA between 24 and 120 h after hepatectomy compared to controls. Similarly, insulin significantly increased the expression of Flt-1 between 24 and 96 h. However, insulin had no significant effect on Flk-1. Furthermore, the immunohistochemical staining revealed that expression of VEGF protein increased in the insulin groups. Insulin significantly increased the PCNA LI of hepatocytes and SECs compared to controls. CONCLUSION: Exogenous insulin may promote SEC proliferation with an enhanced expression of VEGF and its receptor Flt-1 in regenerating rat liver after PHx. PMID- 16273610 TI - Sealing of the hepatic resection area using fibrin glue reduces significant amount of postoperative drain fluid. AB - AIM: To investigate whether the routine use of fibrin glue applied onto the hepatic resection area can diminish postoperative volume of bloody or biliary fluids drained via intraoperatively placed perihepatic tubes and can thus lower the complication rate. METHODS: Two groups of consecutive patients with a comparable spectrum of recent hepatic resections were compared: (1) 13 patients who underwent application of fibrin glue immediately after resection of liver parenchyma; (2) 12 patients who did not. Volumes of postoperative drainage fluid were determined in 4-h intervals through 24 h indicating the intervention caused bloody and biliary segregation. RESULTS: Through the first 8 h postoperatively, there was a tendency of higher amounts of fluids in patients with no additional application of fibrin glue while through the following intervals, a significant increase of drainage volumes was documented in comparison with the first two 4-h intervals, e.g., after 12 h, 149.6 mL +/-110 mL vs 63.2 mL +/-78 mL. Using fibrin glue, postoperative fluid amounts were significantly lower through the postoperative observation period of 24 h (851 mL +/-715 mL vs 315 mL +/-305 mL). CONCLUSION: For hepatic resections, the use of fibrin glue appears to be advantageous in terms of a significant decrease of surgically associated segregation of blood or bile out of the resection area. This might result in a better outcome. PMID- 16273611 TI - Low circulating levels of gastrin-17 in patients with Barrett's esophagus. AB - AIM: To examine whether the fasting levels of serum gastrin-17 (G-17) are lower in Barrett's esophagus (BE) patients than in non-Barrett controls. METHODS: Nineteen patients with BE (presenting with a tubular segment > or =2 cm long in lower esophagus and intestinal metaplasia of incomplete type ("pecialized columnar epithelium") in endoscopic biopsies from the tubular segment below the squamocolumnar junction were collected prospectively from outpatients referred to diagnostic gastroscopy. The controls comprised 199 prospectively collected dyspeptic outpatients without BE or any endoscopically visible lesions in the upper GI tract. Fasting levels of serum G-17 (G-17fast) were assayed with an EIA test using a mAb highly specific to amidated G-17. None of the patients and controls received therapy with PPIs or other antisecretory agents. RESULTS: The mean and median levels of G-17fast in serum were significantly lower (P = 0.001) in BE patients than in controls. The positive likelihood ratios (LR+) of low G 17fast to predict BE in the whole study population at G-17fast levels <0.5, <1, or <1.5 pmol/L were 3.5, 3.0, and 2.8, respectively. Among patients and controls with healthy stomach mucosa, the LR+ were 5.6, 3.8, and 2.6, respectively. In the whole study population, serum G-17 was below 2 pmol/L in 15 of 19 BE patients (79%). The corresponding prevalence was 66 of 199 (33%) in controls (P<0.001). The G-17fast was 5 pmol/L or more in only one of the 19 BE patients (5%). In controls, 76 of the 199 patients (38%) had such high serum G-17fast levels (P<0.01). CONCLUSION: Serum levels of G-17fast tend to be lower in native patients with BE than in healthy controls. PMID- 16273612 TI - Expression profiling of gastric cancer samples by oligonucleotide microarray analysis reveals low degree of intra-tumor variability. AB - AIM: Gene expression profiling provides an unique opportunity to gain insight into the development of different types of gastric cancer. Tumor sample heterogeneity is thought to decrease the sensitivity and tumor specificity of microarray analysis. Thus, microdissection and pre-amplification of RNA is frequently performed. However, this technique may also induce considerable changes to the expression profile. To assess the effect of gastric tumor heterogeneity on expression profiling results, we measured the variation in gene expression within the same gastric cancer sample by performing a gene chip analysis with two RNA preparations extracted from the same tumor specimen. METHODS: Tumor samples from six intestinal T2 gastric tumors were dissected under liquid nitrogen and RNA was prepared from two separate tumor fragments. Each extraction was individually processed and hybridized to an Affymetrix U133A gene chip covering approximately 18 000 human gene transcripts. Expression profiles were analyzed using Microarray Suite 5.0 (Affymetrix) and GeneSpring 6.0 (Silicon Genetics). RESULTS: All gastric cancers showed little variance in expression profiles between different regions of the same tumor sample. In this case, gene chips displayed mean pair wise correlation coefficients of 0.94+/-0.02 (mean+/ SD), compared to values of 0.61+/-0.1 for different tumor samples. Expression of the variance between the two expression profiles as a percentage of "total change" (Affymetrix) revealed a remarkably low average value of 1.18+/-0.78 for comparing fragments of the same tumor sample. In contrast, comparison of fragments from different tumors revealed a percentage of 24.4+/-4.5. CONCLUSION: Our study indicates a low degree of expression profile variability within gastric tumor samples isolated from one patient. These data suggest that tumor tissue heterogeneity is not a dominant source of error for microarray analysis of larger tumor samples, making total RNA extraction an appropriate strategy for performing gene chip expression profiling of gastric cancer. PMID- 16273613 TI - Effect of herpesvirus infection on pancreatic duct cell secretion. AB - AIM: To examine the effect of acute infection caused by herpesvirus (pseudorabies virus, PRV) on pancreatic ductal secretion. METHODS: The virulent Ba-DupGreen (BDG) and non-virulent Ka-RREp0lacgfp (KEG) genetically modified strains of PRV were used in this study and both of them contain the gene for green fluorescent protein (GFP). Small intra/interlobular ducts were infected with BDG virus (10(7) PFU/mL for 6 h) or with KEG virus (10(10) PFU/mL for 6 h), while non-infected ducts were incubated only with the culture media. The ducts were then cultured for a further 18 h. The rate of HCO(3)(-) secretion (base efflux -J(B-)) was determined from the buffering capacity of the cells and the initial rate of intracellular acidification (1) after sudden blockage of basolateral base loaders with dihydro-4,4-diisothiocyanatostilbene-2,2-disulfonic acid (500 micromol/L) and amiloride (200 micromol/L), and (2) after alkali loading the ducts by exposure to NH(4)Cl. All the experiments were performed in HCO(3)(-)-buffered Ringer solution at 37 degrees (n = 5 ducts for each experimental condition). Viral structural proteins were visualized by immunohistochemistry. Virally encoded GFP and immunofluorescence signals were recorded by a confocal laser scanning microscope. RESULTS: The BDG virus infected the majority of accessible cells of the duct as judged by the appearance of GFP and viral antigens in the ductal cells. KEG virus caused a similarly high efficiency of infection. After blockage of basolateral base loaders, BDG infection significantly elevated -J(B-) 24 h after the infection, compared to the non-infected group. However, KEG infection did not modify -J(B-). After alkali loading the ducts, -J(B-) was significantly elevated in the BDG group compared to the control group 24 h after the infection. As we found with the inhibitor stop method, no change was observed in the group KEG compared to the non-infected group. CONCLUSION: Incubation with the BDG or KEG strains of PRV results in an effective infection of ductal epithelial cells. The BDG strain of PRV, which is able to initiate a lytic viral cycle, stimulates HCO(3)(-) secretion in guinea pig pancreatic duct by about four to fivefold, 24 h after the infection. However, the KEG strain of PRV, which can infect, but fails to replicate, has no effect on HCO(3)(-) secretion. We suggest that this response of pancreatic ducts to virulent PRV infection may represent a defense mechanism against invasive pathogens to avoid pancreatic injury. PMID- 16273615 TI - Helicobacter pylori antibiotic resistance in Iran. AB - AIM: To examine the frequency of antibiotic resistance in Iranian Helicobacter pylori (H pylori) strains isolated from two major hospitals in Tehran. METHODS: Examination of antibiotic resistance was performed on 120 strains by modified disc diffusion test and PCR-RFLP methods. In addition, in order to identify the possible causes of the therapeutic failure in Iran, we also determined the resistance of these strains to the most commonly used antibiotics (metronidazole, amoxicillin, and tetracycline) by modified disc diffusion test. RESULTS: According to modified disc diffusion test, 1.6% of the studied strains were resistant to amoxicillin, 16.7% to clarithromycin, 57.5% to metronidazole, and there was no resistance to tetracycline. Of the clarithromycin resistant strains, 73.68% had the A2143G mutation in the 23S rRNA gene, 21.05% A2142C, and 5.26% A2142G. None of the sensitive strains were positive for any of the three point mutations. Of the metronidazole resistant strains, deletion in rdxA gene was studied and detected in only 6 (5%) of the antibiogram-based resistant strains. None of the metronidazole sensitive strains possessed rdxA gene deletion. CONCLUSION: These data show that despite the fact that clarithromycin has not yet been introduced to the Iranian drug market as a generic drug, nearly 20% rate of resistance alerts toward the frequency of macrolide resistance strains, which may be due to the widespread prescription of erythromycin in Iran. rdxA gene inactivation, if present in Iranian H pylori strains, may be due to other genetic defects rather than gene deletion. PMID- 16273616 TI - Receptor-binding cancer antigen expressed on SiSo cells can be detected in metastatic lymph nodes from gastrointestinal cancers. AB - AIM: To investigate the expression of receptor-binding cancer antigen expressed on SiSo cells (RCAS1) in metastatic lymph nodes from gastrointestinal cancer. METHODS: Metastatic lymph nodes from gastrointestinal cancer were detected for RCAS1 by immunohistochemical staining and mRNA in situ hybridization. RESULTS: A total of 102 metastatic lymph nodes from bile duct, gastric, colon and pancreatic cancer were investigated for RCAS1 expression. The immunoreactivity of RCAS1 was identified in 100% of metastatic lymph nodes. Both local and distant metastatic lymph nodes showed RCAS1 expression. On the contrary, specimens of non-cancerous lymph nodes were negative for RCAS1. The result of mRNA in situ hybridization was also confirmed by the finding of immunohistochemical staining. RCAS1 mRNA was detected in all tumor cells that metastasized to lymph nodes. CONCLUSION: All metastatic lymph nodes express RCAS1 in tumor cells at both mRNA levels, and RCAS1 that should be used as a complementary factor for identification of metastatic lymph nodes from gastrointestinal cancers. PMID- 16273614 TI - Haplotype of prostaglandin synthase 2/cyclooxygenase 2 is involved in the susceptibility to inflammatory bowel disease. AB - AIM: Prostaglandin G/H synthase 2 (PTGS2 or COX2) is one of the key factors in the cellular response to inflammation. PTGS2 is expressed in the affected intestinal segments of patients with inflammatory bowel diseases (IBD). In IBD patients, non-steroidal anti-inflammatory drugs, which have been shown to reduce both the production and activity of PTGS2, may activate IBD and aggravate the symptoms. We aimed at examining genetic variants of PTGS2 that may be risk factors for IBD. METHODS: We have genotyped 291 individuals diagnosed with IBD and 367 controls from the Dutch population for the five most frequent polymorphisms of the PTGS2 gene. Clinical data were collected on all patients. DNA was extracted via normal laboratory methods. Genotyping was carried out using multiplex PCR followed by the Invader Assay and the 5' exonuclease assay (TaqMan). New polymorphism screening was performed by pre-screening with denaturing high-performance liquid chromatography, followed by fluorescent sequencing. RESULTS: Allele 5209G was weakly associated with Crohn's disease (odds ratio (OR) 1.63, 95% confidence interval (CI) 1.03-2.57), and allele 8473T with ulcerative colitis (OR 1.50, 95%CI 1.00-2.27). The haplotype including both alleles showed a strong association with IBD (OR 13.15, 95%CI 3.17-116.15). This haplotype, while rare (-0.3%) in the general population, is found more frequently in the patients (3.5%). CONCLUSION: Our data suggest that this haplotype of PTGS2 contributes to the susceptibility of IBD. PMID- 16273617 TI - Normal serum alanine aminotransferase activity in uncomplicated obesity. AB - AIM: To evaluate serum alanine aminotransferase (ALT) activity in a well characterized group of uncomplicated obese subjects and its correlation with insulin resistance, plasma adiponectin, and leptin concentrations. METHODS: One hundred and five uncomplicated obese subjects (87 women, 18 men, age 34.3+/-9.6 years, BMI 39.9+/-8.3 kg/m(2)) were studied. Serum ALT activity was evaluated. Insulin sensitivity was assessed by euglycemic hyperinsulinemic clamp (M index) and fasting insulin. Plasma leptin and adiponectin levels were also measured. RESULTS: Serum ALT concentration in the whole group of uncomplicated obese subjects was 17.73+/-6.33 U/L with none of the subjects presenting ALT levels greater than 43 U/L and only 9 (11%) women and 3 (19%) men showed ALT levels >19 and >30 U/L for women and men, respectively. No significant difference was detected in serum ALT levels between severe obese subjects (BMI >40 kg/m(2)) and those with BMI <40 kg/m(2) (18.63+/-6.25 vs 17.26+/-6.02 U/L). ALT was significantly correlated with fasting insulin (r = 0.485, P = 0.02) and triglycerides (r = 0.358, P = 0.03). CONCLUSION: Serum ALT activity is practically normal in uncomplicated obese subjects, independently of their obesity degree. These findings suggest the role of obesity-related comorbidities and not of BMI as main risk factors for elevated ALT levels in obese subjects. PMID- 16273618 TI - Endoscopic treatment and follow-up of gastrointestinal Dieulafoy's lesions. AB - AIM: To investigate retrospectively the clinical and endoscopic features of bleeding Dieulafoy's lesions and to assess the short- and long-term effectiveness of endoscopic treatment. METHODS: Twenty-three patients who had gastrointestinal bleeding from Dieulafoy's lesions underwent endoscopic therapy. Demographic data, mode of presentation, risk factors for gastrointestinal bleeding, blood transfusion requirements, endoscopic findings, details of endoscopic therapy, recurrence of bleeding and mortality rates were collected and analyzed retrospectively. RESULTS: Hemostasis was attempted by dextrose 50% plus epinephrine in 10 patients, hemoclipping in 8 patients, heater probe in 2 patients and ethanolamine oleate in 2 patients. Comorbid conditions were present in 17 patients (74%). Overall permanent hemostasis was achieved in 18 patients (78%). Initial hemostasis was successful with no recurrent bleeding in patients treated with hemoclipping, heater probe or ethanolamine injection. In the group of patients who received dextrose 50% plus epinephrine injection treatment, four (40%) had recurrent bleeding and one (10%) had unsuccessful initial hemostasis. Of the four patients who had rebleeding, three had unsuccessful hemostasis with similar treatment. Surgical treatment was required in five patients (22%) owing to uncontrolled bleeding, recurrent bleeding with unsuccessful retreatment and inability to approach the lesion. One patient (4.3%) died of sepsis after operation during hospitalization. There were no side-effects related to endoscopic therapy. None of the patients in whom permanent hemostasis was achieved presented with rebleeding from Dieulafoy's lesion over a mean long-term follow-up of 29.8 mo. CONCLUSION: Bleeding from Dieulafoy's lesions can be managed successfully by endoscopic methods, which should be regarded as the first choice. Endoscopic hemoclipping therapy is recommended for bleeding Dieulafoy's lesions. PMID- 16273619 TI - Manometric assessment of idiopathic megarectum in constipated children. AB - AIM: Chronic constipation is a frequent finding in children. In this age range, the concomitant occurrence of megarectum is not uncommon. However, the definition of megarectum is variable, and a few data exist for Italy. We studied anorectal manometric variables and sensation in a group of constipated children with megarectum defined by radiologic criteria. Data from this group were compared with those obtained in a similar group of children with recurrent abdominal pain. METHODS: Anorectal testing was carried out in both groups by standard manometric technique and rectal balloon expulsion test. RESULTS: Megarectum patients displayed discrete abnormalities of anorectal variables and sensation with respect to controls. In particular, the pelvic floor function appeared to be impaired in most patients. CONCLUSION: Constipated children with megarectum have abnormal anorectal function and sensation. These findings may be helpful for a better understanding of the pathophysiological basis of this condition. PMID- 16273620 TI - Different effects of a CD14 gene polymorphism on disease outcome in patients with alcoholic liver disease and chronic hepatitis C infection. AB - AIM: Clinical and experimental data suggest that gut-derived endotoxins are an important pathogenic factors for progression of chronic liver disease. Recently, a C-T (-159) polymorphism in the promoter region of the CD14 gene was detected and found to confer increased CD14 expression and to be associated with advanced alcoholic liver damage. Here, we investigated this polymorphism in patients with less advanced alcoholic liver disease (ALD) and chronic hepatitis C virus (HCV) infection. METHODS: CD14 genotyping was performed by PCR-RFLP analysis in (a) 121 HCV patients, (b) 62 patients with alcohol-associated cirrhosis (Alc-Ci), (c) 118 individuals with heavy alcohol abuse without evidence of advanced liver damage (Alc-w/o Ci), and (d) 247 healthy controls. Furthermore, serum levels of soluble CD14 (sCD14) and transaminases were determined. RESULTS: The TT genotype was significantly more frequent in Alc-Ci compared to Alc-w/o Ci or controls (40.3% vs 23.7% or 24.0%, respectively). In Alc-w/o Ci, serum levels of transaminases did not differ significantly between patients with different CD14 genotypes. In HCV patients, TT-homozygotes had significantly higher sCD14 levels and sCD14 serum levels were significantly higher in patients with advanced fibrosis or cirrhosis. However, no association was found between CD14 genotypes and histological staging or grading. CONCLUSION: Considering serum transaminases as surrogate markers for alcoholic liver damage, the CD14 polymorphism seems to exhibit different effects during the course of ALD. Differences in genotype distribution between cirrhotic HCV patients and alcoholics and the known functional impact of this polymorphism on CD14 expression levels further indicate differences in the pathophysiological role of CD14 and CD14-mediated lipopolysaccharides signal transduction with regard to the stage as well as the type of the underlying liver disease. PMID- 16273621 TI - Hyperhomocysteinemia in ulcerative colitis is related to folate levels. AB - AIM: To study the prevalence and clinical significance of hyperhomocysteinemia (hHcys), an independent factor for arterial and venous thrombosis, in a group of patients with ulcerative colitis (UC). METHODS: Fasting homocysteine (Hcys), folate, and vitamin B(12) serum levels were measured in 40 UC patients and 50 healthy controls. Clinical data regarding UC were gathered. RESULTS: Median serum Hcys levels in UC patients were similar to controls (12.26 micromol/L vs 12.32 micromol/L), but the prevalence of hHcys was higher in UC patients than in controls (30% vs 10%, P = 0.028). UC significantly increased the risk of hHcys (adjusted odds ratio: 4.125; 95%CI: 1.26-13.44). Multivariate regression analysis showed that male sex, folate and vitamin B(12) deficiency or lower serum values were significant independent predictors of higher Hcys levels in UC patients (r(2) = 0.4; P<0.001). CONCLUSION: HHcys is common in UC patients and it is related to folate and vitamin B(12) deficiency or lower serum values. It would be reasonable for patients with UC to receive folate and vitamin B complex supplements as a prophylactic measure. PMID- 16273622 TI - CD14 promoter polymorphism in Chinese alcoholic patients with cirrhosis of liver and acute pancreatitis. AB - AIM: To investigate the relationship between genetic polymorphism of the CD14 promoter and the occurrence of alcoholic cirrhosis and alcoholic pancreatitis, and to challenge the conclusion made earlier that the patients with acute alcoholic pancreatitis and patients with alcoholic cirrhosis of liver are two different subpopulations. METHODS: Using the polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) method, we determined the polymorphism of CD14 gene and aldehyde dehydrogenase gene 2 (ALDH 2) in 335 alcoholic patients with different organ complications i.e., cirrhosis of liver (n = 100), acute pancreatitis (n = 100), esophageal cancer (n = 82) and avascular necrosis of hip joint (AVN) (n = 53) and 194 non-alcoholic controls in a Chinese group. RESULTS: The results showed that the carriage of T allele was not different among alcoholic patients with cirrhosis of liver, alcoholic patients with other complication and non-alcoholic controls. On the other hand, the carriage of the C allele was significantly more prevalent for alcoholic pancreatitis than for esophageal cancer (0.79 vs 0.60, P<0.001), alcoholic AVN (0.79 vs 0.65, P<0.025) and non-alcoholic controls (0.79 vs 0.68, P<0.025). Furthermore, when only subjects with ALDH2 1-1 genotype were examined, the C allele frequency was significantly more prevalent for alcoholic pancreatitis than for alcoholic liver cirrhosis (0.82 vs 0.69, P<0.025), esophageal cancer (0.82 vs 0.61, P<0.01), alcoholic AVN (0.82 vs 0.64, P<0.01) and non-alcoholic controls (0.82 vs 0.69, P<0.05). CONCLUSION: The C allele may be associated with some mechanism, which is important in the pathogenesis of alcoholic pancreatitis, and that alcoholic patients with acute pancreatitis and cirrhosis of liver are probably two different subpopulations. PMID- 16273623 TI - Balthazar computed tomography severity index is superior to Ranson criteria and APACHE II scoring system in predicting acute pancreatitis outcome. AB - AIM: Acute pancreatitis (AP) is a process with variable involvement of regional tissues or organ systems. Multifactorial scales included the Ranson, Acute Physiology and Chronic Health Evaluation (APACHE II) systems and Balthazar computed tomography severity index (CTSI). The purpose of this review study was to assess the accuracy of CTSI, Ranson score, and APACHE II score in course and outcome prediction of AP. METHODS: We reviewed 121 patients who underwent helical CT within 48 h after onset of symptoms of a first episode of AP between 1999 and 2003. Fourteen inappropriate subjects were excluded; we reviewed the 107 contrast enhanced CT images to calculate the CTSI. We also reviewed their Ranson and APACHE II score. In addition, complications, duration of hospitalization, mortality rate, and other pathology history also were our comparison parameters. RESULTS: We classified 85 patients (79%) as having mild AP (CTSI <5) and 22 patients (21%) as having severe AP (CTSI > or =5). In mild group, the mean APACHE II score and Ranson score was 8.6+/-1.9 and 2.4+/-1.2, and those of severe group was 10.2+/-2.1 and 3.1+/-0.8, respectively. The most common complication was pseudocyst and abscess and it presented in 21 (20%) patients and their CTSI was 5.9+/-1.4. A CTSI > or =5 significantly correlated with death, complication present, and prolonged length of stay. Patients with a CTSI > or =5 were 15 times to die than those CTSI <5, and the prolonged length of stay and complications present were 17 times and 8 times than that in CTSI <5, respectively. CONCLUSION: CTSI is a useful tool in assessing the severity and outcome of AP and the CTSI > or =5 is an index in our study. Although Ranson score and APACHE II score also are choices to be the predictors for complications, mortality and the length of stay of AP, the sensitivity of them are lower than CTSI. PMID- 16273624 TI - Downregulation of alpha-fetoprotein siRNA inhibits proliferation of SMMC-7721 cells. AB - AIM: To study the function of alpha-fetoprotein (AFP) in SMMC-7721 hepatoma cells. METHODS: A hairpin siRNA expressing plasmid pSilencer3.0-H1-afp was constructed and transfected into SMMC-7721 cells with Lipofectamine 2000. The expression of AFP was monitored by real-time RT-PCR and immunoassays, its effect on SMMC-7721 cell proliferation and cell death was detected by MTT and fluorescence activated cell sorter (FACS). RESULTS: The AFP-siRNA expressing plasmid downregulated the expression of AFP obviously (about 34%), and inhibited SMMC-7721 cell proliferation, but did not induce apoptosis. CONCLUSION: Downregulation of AFP siRNA inhibits proliferation of SMMC-7721 cells, but cannot cause apoptosis. PMID- 16273625 TI - Interaction models of CYP1A1, GSTM1 polymorphisms and tobacco smoking in intestinal gastric cancer. AB - AIM: To explore the interaction models of the cytochrome P-450 (CYP) 1A1 Val variant and glutathione S-transferase (GST) M1 null polymorphisms with tobacco smoking in the occurrence of intestinal gastric cancer. METHODS: A community based case-control study was conducted in Yangzhong. Subjects included 114 intestinal types of gastric cancer with endoscopic and pathological diagnosis during January 1997 and December 1998, and 693 controls selected from their spouse, siblings or siblings-in-law who had no history of digestive system cancer. Logistic regression was used to estimate the interaction models. RESULTS: The frequency of the CYP1A1 Val variant allele in cases did not differ from that in controls. The OR of GSTM1 null genotype was 2.0 (95% confidence interval (95%CI): 1.2-3.1, P<0.01). It showed a significant type 2 form of interaction model when both CYP1A1 Val variant allele and former tobacco smoking existed (i.e., among the multiplicative effects, the disease risk is increased by the tobacco exposure alone but not by the CYP1A1 variant alone). The interaction index gamma was 2.8, and OR(eg) (95%CI) was 5.0 (1.9-13.4). GSTM1 null genotype and former tobacco smoking were significant in a type 4 interaction model (i.e., the disease risk is increased by GSTM1 null genotype or tobacco exposure alone among the multiplicative effects). The interaction index gamma and OR(eg) (95%CI) were 3.4 and 8.4 (3.4-20.9), respectively. CONCLUSION: Different interaction models of CYP1A1 Val variant allele and GSTM1 null genotype with tobacco smoking will contribute to understanding carcinogenic mechanism, but there is a need to further investigate in larger scale studies. PMID- 16273627 TI - Multiseptate gallbladder with anomalous pancreaticobiliary ductal union: a case report. AB - Multiseptate gallbladder, characterized by the presence of multiple septa dividing the gallbladder lumen, is a very extremely rare congenital anomaly of the gallbladder. On the other hand, anomalous pancreaticobiliary ductal union is also one of the congenital anomalous biliary diseases and thought to be related with choledochal cyst or biliary tract malignancies. In this paper, we describe a unique and first patient of multiseptate gallbladder with anomalous pancreaticobiliary ductal union and a review of the literature. To clarify more characters of the multiseptate gallbladder, examination of a larger patient population will be needed and further studies will be required. PMID- 16273626 TI - Effect of peroxisome proliferator-activated receptor-gamma ligand on inflammation of human gallbladder epithelial cells. AB - AIM: To investigate the effect of peroxisome proliferator-activated receptor gamma (PPAR-gamma) and its ligand, ciglitazone, on inflammatory regulation of human gallbladder epithelial cells (HGBECs) and to assess the effect of human epithelial growth factor (hEGF) on growth of HGBECs. METHODS: HGBECs were cultured in media containing hEGF or hEGF-free media. HGBECs were divided into normal control group, inflammatory control group and ciglitazone group (test group). Inflammatory control group and ciglitazone group were treated with 5 microg/L of human interleukin-1beta (hIL-1beta) to make inflammatory model of HGBECs. The ciglitazone group was treated with various concentrations of ciglitazone, a potent ligand of PPAR-gamma. Subsequently, interleukin-8 (IL-8), IL-6, and tumor necrosis factor-alpha (TNF-alpha) concentrations in all groups were measured. The data were analyzed statistically. RESULTS: HGBECs were cultured in medium successfully. The longevity of HGBECs in groups containing hEGF was longer than that in hEGF-free groups. So was the number of HGBECs. The longest survival time of HGBEC was 25 d. The inflammatory model of HGBECs was obtained by treating with hIL-1beta. The concentrations of IL-6 and IL-8 in ciglitazone group were lower than those in inflammatory control group (P<0.05). The secretion of IL-6 in inflammatory control group was higher (350.31+/-37.05 microg/L) than that in normal control group (50.0+/-0.00 microg/L, P<0.001). Compared to normal control group, IL-8 concentration in inflammatory control was higher (P<0.05). CONCLUSION: hEGF improves the growth of HGBECs in vitro. Ciglitazone inhibits the inflammation of HGBECs in vitro and has potential therapeutic effect on cholecystitis in vivo. PMID- 16273628 TI - Autoimmune hepatitis triggered by acute hepatitis A. AB - The patient was a 57-year-old woman presenting with jaundice as the chief complaint. She began vomiting on July 10, 2003. Jaundice was noted and admitted to our hospital for thorough testing. Tests on admission indicated severe hepatitis, based on: aspartate aminotransferase (AST), 1 076 IU/L; alanine aminotransferase (ALT), 1 400 IU/L; total bilirubin (TB), 20.9 mg/dL; and prothrombin time rate (PT%), 46.9%. Acute hepatitis A (HA) was diagnosed based on negative hepatitis B surface antigen and hepatitis C virus RNA and positive immunoglobulin (Ig) M HA antibody, but elevation of anti-nuclear antigen (X320) and IgG (3 112 mg/dL) led to suspicion of autoimmune hepatitis (AIH). Plasma exchange was performed for 3 d from July 17, and steroid pulse therapy was performed for 3 d starting on July 18, followed by oral steroid therapy. Liver biopsy was performed on August 5, and the results confirmed acute hepatitis and mild chronic inflammation. Levels of AST and ALT normalized, so dose of oral steroid was markedly reduced. Steroid therapy was terminated after 4 mo, as the patient had glaucoma. Starting 3 mo after cessation of steroid therapy, levels of AST and ALT began to increase again. Another liver biopsy was performed and AIH was diagnosed based on serum data and biopsy specimen. Oral steroid therapy was reinitiated. Levels of AST and ALT again normalized. The present case was thus considered to represent AIH triggered by acute HA. PMID- 16273629 TI - Role of von Willebrand factor levels in the prognosis of stage IV colorectal cancer: do we have enough evidence? PMID- 16273630 TI - Research progress of vasculopathy in portal hypertension. AB - Portal hypertension, one of the vascular diseases, not only has lesions in liver, but also changes in vascular structures and functions of extrahepatic portal system, systemic system and pulmonary circulation. The pathological changes of vasculopathy in portal hypertension include remodeling of arterialized visceral veins, intimal injury of visceral veins and destruction of contractile structure in visceral arterial wall. The mechanisms of vasculopathy in portal hypertension may be attributed to the changes of hemodynamics in portal system, immune response, gene modulation, vasoactive substances, and intrahepatic blood flow resistance. Portal hypertension can cause visceral hyperdynamic circulation, and the development and progression of visceral vasculopathy, while visceral vasculopathy can promote the development and progression of portal hypertension and visceral hyperdynamic circulation in turn. The aforementioned three factors interact in the pathogenesis of hepatic cirrhosis-induced portal hypertension and are involved in hemorrhage due to varicose vein rupture. PMID- 16273631 TI - Chronic hepatitis C virus infection and post-liver transplantation diabetes mellitus. AB - Patients with chronic hepatitis C virus (HCV) infection have a significantly increased prevalence of type 2 diabetes mellitus compared to controls or HBV infected patients. Moreover, the incidence rate of post-liver transplantation diabetes mellitus (PTDM) also appears to be higher among patients with HCV infection. PTDM is often associated with direct viral infection, autoimmune disorders, and immunosuppressive regimen. Activation of tumor necrosis factor alpha may be the link between HCV infection and diabetes. In this article, we reviewed the epidemiologic association between HCV infection and PTDM, highlighting the most recent pathophysiologic insights into the mechanisms underlying this association. PMID- 16273632 TI - Value of endoscopic methylene blue and Lugol's iodine double staining and detection of GST-Pi and telomerase in the early diagnosis of esophageal carcinoma. AB - AIM: To explore the expressions of GST-Pi and telomerase activity in esophageal carcinoma and premalignant lesions and to investigate the value of endoscopic methylene blue (MB) and Lugol's iodine double staining. METHODS: Seventy-two patients with esophagopathy were sprayed endoscopically with MB and Lugol's iodine in proper order and the areas stained blue and brown, and the area between the blue and brown stains were obtained. Depending on the pattern of mucosal staining, biopsy specimen was obtained. GST-Pi and telomerase activity in specimens were examined by immunohistochemistry and PCR-based silver staining telomeric repeat amplification protocol, respectively. RESULTS: After MB and Lugol's iodine staining, the area between both the colors was obtained in 64 of the 72 patients and the areas were stained blue and brown in all of the 72 patients. Association test of two simultaneous ordinal categorical data showed a correlation between the esophageal mucosal staining and the esophageal histology (P<0.005). The expression of GST-Pi and telomerase activity in esophageal carcinoma and premalignant lesions increased. The expression of GST-Pi and telomerase activity in dysplasia and carcinoma was significantly higher than that in normal epithelium (P<0.005). The expression in hyperplasia was slightly higher than that in normal epithelium. With the lesions progressing from low- to moderate- to high-grade dysplasia, the positive rate increased (P<0.025). Expression of GST-Pi was correlated with that of telomerase activity in dysplasia and carcinoma (phis = 0.4831, P<0.005; phis = 0.3031, P<0.025, respectively); but there was no correlation between them in normal epithelium and hyperplasia. CONCLUSION: The expression of GST-Pi and telomerase may be an early event in the carcinogenesis of esophagus. They may play an induced and synergistic role with each other in the carcinogenesis of esophagus. Endoscopic MB and Lugol's iodine double staining and detection of GST-Pi and telomerase activity may contribute to the early diagnosis of esophageal carcinoma. PMID- 16273633 TI - Epstein-Barr virus-associated gastric carcinoma: Evidence of age-dependence among a Mexican population. AB - AIM: To investigate features of Epstein-Barr virus (EBV)-associated gastric carcinoma (EBVaGC) among a Mexican population. METHODS: Cases of primary gastric adenocarcinoma were retrieved from the files of the Departments of Pathology at the Instituto Nacional de Cancerologia and the Instituto Nacional de la Nutricion in Mexico City. The anatomic site of the gastric neoplasia was identified, and carcinomas were histologically classified as intestinal and diffuse types and subclassified as proposed by the Japanese Research Society for Gastric Cancer. EBV-encoded small non-polyadenylated RNA-1 (EBER-1) in situ hybridization was conducted to determine the presence of EBV in neoplastic cells. RESULTS: We studied 330 consecutive, non-selected, primary gastric carcinomas. Among these, there were 173 male and 157 female patients (male/female ratio 1.1/1). EBER-1 was detected in 24 (7.3%) cases (male/female ratio: 1.2/1). The mean age for the entire group was 58.1 years (range: 20-88 years), whereas the mean age for patients harboring EBER-1-positive gastric carcinomas was 65.3 years (range: 50 84 years). Age and histological type showed statistically significant differences, when EBER-1-positive and -negative gastric carcinomas were compared. EBER-1 was detected in hyperplastic- and dysplastic-gastric mucosa surrounding two EBER-1-negative carcinomas, respectively. CONCLUSION: Among Latin-American countries, Mexico has the lowest frequency of EBVaGC. Indeed, the Mexican population >50 years of age was selectively affected. Ethnic variations are responsible for the epidemiologic behavior of EBVaGC among the worldwide population. PMID- 16273635 TI - Effects of adenovirus-mediated human cyclooxygenase-2 antisense RNA on the growth of hepatocellular carcinoma. AB - AIM: To investigate the relation between the expression of cyclooxygenase-2 (COX 2) and liver cancer, to construct the recombinant adenovirus encoding human COX-2 antisense RNA, and to explore its effects on liver cancer cell proliferation. METHODS: We studied the expression of COX-2 in 34 cases of hepatocellular carcinoma (HCC) and SMMC7402 and SMMC7721 by immunohistochemical technique. Recombinant adenovirus Ad-AShcox-2 was constructed and transfected into human HCC cell lines SMMC7402 and SMMC7721, and its effects on COX-2 expression, cell apoptosis and cell cycle were analyzed by flow cytometry. Cell proliferation was determined by colony-forming efficiency. RESULTS: We observed COX-2 expression in 82.4% of the HCC and SMMC7402 cells, but no COX-2 expression in SMMC7721 cells. In addition, recombinant adenovirus encoding antisense COX-2 fragment Ad-AShcox-2 was obtained with the titer of 1.06 x 10(12) PFU/mL. Ad-AShcox-2 could reduce the expression of COX-2 and enhance the percentage of cells in G(1)/G(0) phase in SMMC7402 cell line. The difference of apoptotic index between the Ad-AShcox-2 group and control group was statistically significant (t( control group ) = 32.62 and t( Ad-LacZ ) = 10.93, P<0.001) in SMMC7402 but not in SMMC7721. Similarly, colony-forming rates of SMMC7402 and SMMC7721 cell lines, after the transfer of Ad-AShcox-2, were (2.7+/-0.94)% and (33.6+/-4.24)%, respectively. CONCLUSION: Reduction in the expression of COX-2 can inhibit COX-2 expressing HCC cells. PMID- 16273634 TI - Combined TACE and PEI for palliative treatment of unresectable hepatocellular carcinoma. AB - AIM: To assess whether the effectiveness of a combination of transarterial chemoembolization (TACE) and percutaneous ethanol injection (PEI) in the treatment of unresectable hepatocellular carcinoma (HCC) is superior to TACE alone a randomized controlled trial was performed. METHODS: The effect of combination therapy on long-term survival rates and duration of hospitalization was evaluated in 52 previously untreated HCCs, randomly allocated to TACE-PEI (27 pts) or TACE alone (25 pts). RESULTS: The cumulative survival rate of the TACE group was 75.8% at 6 mo, 62.9% at 12 mo, and 18.0% at 24 mo and of the TACE-PEI group 76.9%, 61.5%, and 38.7%, respectively. Comparison of overall survival in both groups showed no statistically significant difference. Regarding the patients with HCCs Okuda stage I (n = 26), the median survival of the TACE-PEI group was significantly longer (>24 mo, median not yet reached in the study period) compared to the TACE group (18.4 mo [range 11.6-21.7 mo]; P = 0.04). TACE PEI reduced the relative risk for mortality to 0.4 (95%CI 0.15-0.96) compared to patients who received TACE alone. Median survival in patients with HCCs Okuda stage II or III was 5.0 mo in the TACE group (1.7 mo-not defined) compared to 10.4 mo in the TACE-PEI group. CONCLUSION: The combination TACE-PEI improved survival time compared to TACE alone. Our study revealed a statistically significant improved survival in HCCs Okuda stage I. Side effects were minor and the combination therapy did not prolong duration of hospitalization considerably. PMID- 16273636 TI - Usefulness of serum des-gamma-carboxy prothrombin in detection of hepatocellular carcinoma. AB - AIM: Des-gamma-carboxy prothrombin (DCP) has been reported to be more sensitive and specific in diagnosing hepatocellular carcinoma (HCC) when compared with alpha-fetoprotein (AFP). However, its ability to identify small HCC still remains unclear. Thus, we conducted a cross-sectional case control study to evaluate whether DCP is better than AFP for differentiating HCC from nonmalignant liver disease and further evaluate the usefulness of DCP in early diagnosis of small HCC. METHODS: Serum DCP and AFP levels were determined in 127 patients. Among these patients, 32 were with non-cirrhotic chronic hepatitis, 34 were with compensated cirrhosis, and 61 were with HCC. The cut-off value for the DCP and AFP were set as 40 mAU/mL and 20 ng/mL, respectively. To compare the diagnostic value of DCP and AFP in distinguishing HCC from nonmalignant chronic liver disease, receiver operating characteristic (ROC) curves were constructed for each assay. RESULTS: The accuracy, sensitivity and specificity of DCP were higher than AFP in detecting HCC (81.9%, 77% and 86.4% vs 68.5%, 59% and 77.3%, respectively). The area under the ROC (AUROC) curves revealed that DCP had a better accuracy than AFP in diagnosis of HCC (0.85 [95%CI, 0.78-0.91] vs 0.73 [95%CI, 0.65-0.81], P = 0.013). In 39 patients with solitary HCC, the positive rates of DCP were 100% in patients with tumor size larger than 3 cm, 66.7% in patients with tumor size 2-3 cm and 50% in patients with tumor size less than 2 cm. The positive rates of AFP in patients with tumor size larger than 3 cm, 2-3 cm and less than 2 cm were 55.6%, 50%, and 33.3%, respectively. The median level of DCP in HCC patients with tumor size larger than 3 cm was significantly higher than those with tumor size 2-3 cm and those with the size of less than 2 cm. CONCLUSION: Our study indicates that DCP has a better diagnostic value than AFP in differentiating HCC from nonmalignant chronic liver disease. DCP has not only a stronger correlation with HCC than AFP in tumor size but also more effectiveness than AFP in detecting small size of HCC. PMID- 16273638 TI - Mechanism of counterattack of colorectal cancer cell by Fas/Fas ligand system. AB - AIM: To determine the role of Fas/Fas ligand (FasL) in the immune escape of colon cancer cells. METHODS: Immunohistochemistry was used to observe the expression of Fas and FasL in the tissues of colon cancer patients. In situ hybridization was used to detect the localization of FasL mRNA expression in cancer tissues. Terminal deoxynucleotide transferase-mediated dUTP nick end labeling (TUNEL) assay and CD45 staining were performed to detect the apoptosis of tumor infiltrating lymphocytes (TILs). Co-culture assays of colon cancer cells (SW480) and Jurkat cells (Fas-sensitive cells) were performed to observe the counterattack of colon cancer cells to lymphocytes. RESULTS: Of 53 cases of colon carcinomas, 23 cases (43.4%) expressed Fas which was significantly lower as compared to the normal colonic mucosa (73.3%, P<0.01), and 45 cases (84.9%) of colon carcinomas expressed FasL, whereas only two cases (3.75%) in normal mucosa expressed FasL. FasL expression in the colon cancer cells was found to be associated with increased cell death of TILs. The apoptotic rate of TIL in the FasL-positive staining regions of tumor cells was significantly higher than that in the FasL-negative staining region (54.84+/-2.79% vs 25.73+/-1.98%, P<0.01). The co-culture of SW480 cells and Jurkat cells confirmed the function of FasL on the SW480 cells. The apoptotic rates of Jurkat cells were found to be related with the amount of SW480 cells. CONCLUSION: Colon cancer cells can escape the immune surveillance and killing via decreasing Fas expression, and can counterattack the immune system via increasing FasL expression. Fas/FasL can serve as potential targets for effective antitumor therapy. PMID- 16273637 TI - Genetic alterations and expression of inhibitor of growth 1 in human sporadic colorectal cancer. AB - AIM: To explore the effect and significance of inhibitor of growth 1 (ING1) gene in carcinogenesis and progression of human sporadic colorectal cancer. METHODS: mRNA expression, mutation, and loss of heterozygosity (LOH) of ING1 gene in 35 specimens of sporadic colorectal cancer tissues and the matched normal mucous membrane tissues were detected by semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR), PCR-single strain conformation polymorphism (PCR-SSCP) and PCR-simple sequence length polymorphism (PCR-SSLP) using microsatellite markers, respectively. RESULTS: The average ratios of light intensities of p33(ING1b) and p47(ING1a) mRNA expression in the cancerous tissues were significantly lower than those in normal tissues. The difference between the two mRNA splices was not significant in the matched tissues. In addition, the ratios of light intensities of p33(ING1b) and p47(ING1a) mRNA expression in the cancerous tissues of Dukes' stages C and D were significantly lower than those in cancerous tissues of Dukes' stages A and B. However, no mutation of ING1 gene was detected in all 35 cases; only 4 cases of LOH (11.4%) were found. CONCLUSION: p33(ING1b) and p47(ING1a) mRNA expressions are closely related with the carcinogenesis and progression of human sporadic colorectal cancer. No mutation of ING1 gene is found, and there are only few LOH in sporadic colorectal cancers. These might not be the main reasons for the down regulation of ING1 expression. Its low expression may happen in transcription or post-transcription. PMID- 16273639 TI - Specific ssDNA concentration in liver tissue as an index of apoptosis in hepatitis C virus-infected patients. AB - AIM: To evaluate the activity of apoptosis in liver tissue and explore its possible association with hepatic necroinflammation and fibrosis as well as serum hepatitis C virus (HCV) load. METHODS: The studied population included 50 chronic hepatitis C patients (20 women and 30 men, aged 18-66 years). HCV-RNA quantification was performed by two-step real-time quantitative RT-PCR method using the TaqMan technology (reagents of Applera Corporation firm, USA). The morphology of liver tissue was assessed descriptively and scored (necroinflammatory activity and fibrosis). The early apoptosis activity in liver tissue was examined by ssDNA apoptosis ELISA kit, (Chemicon, Germany). RESULTS: The correlation between apoptosis and fibrosis in liver tissue was observed. High intensification of apoptosis was proportional to the increase of fibrosis (ssDNA: 16.65 x 10(-5) microug/g; 12.71 x 10(-5) microg/g), however, this difference was not statistically significant (P>0.05). Activity of apoptosis in the liver tissue, expressed by ssDNA concentration did not depend on hepatic necroinflammatory changes, HCV-RNA viral load, ALT, and AST activity as well as prothrombin time and INR index. CONCLUSION: Fibrosis in the tissue is closely associated with early apoptosis in HCV-infected patients. PMID- 16273641 TI - Endothelin-1 stimulates contraction and migration of rat pancreatic stellate cells. AB - AIM: Activated pancreatic stellate cells (PSCs) are implicated in the pathogenesis of pancreatic fibrosis and inflammation. Endothelin-1 (ET-1), which acts through G-protein coupled ET(A) and ET(B) receptors, has several biological activities. We here examined the ability of ET-1 to affect the cell functions of PSCs and the underlying molecular mechanisms. METHODS: PSCs were isolated from the pancreas of male Wistar rats after perfusion with collagenase, and cells between passages two and five were used. Expression of ET-1 and ET receptors was assessed by reverse transcription-PCR and immunostaining. Phosphorylation of myosin regulatory light chain (MLC), extracellular-signal regulated kinase (ERK), and Akt was examined by Western blotting. Contraction of PSCs was assessed on hydrated collagen lattices. Cell migration was examined using modified Boyden chambers. Cell proliferation was assessed by measuring the incorporation of 5 bromo-2'-deoxyuridine. RESULTS: Culture-activated PSCs expressed ET(A) and ET(B) receptors, and ET-1. ET-1 induced phosphorylation of MLC and ERK, but not Akt. ET 1 induced contraction and migration, but did not alter proliferation of PSCs. ET 1-induced contraction was inhibited by an ET(A) receptor antagonist BQ-123 and an ET(B) receptor antagonist BQ-788, whereas migration was inhibited by BQ-788 but not by BQ-123. A Rho kinase inhibitor Y-27632 abolished both contraction and migration. CONCLUSION: ET-1 induced contraction and migration of PSCs through ET receptors and activation of Rho-Rho kinase. ET(A) and ET(B) receptors play different roles in the regulation of these cellular functions in response to ET 1. PMID- 16273640 TI - NF-kappaB and ERK-signaling pathways contribute to the gene expression induced by cag PAI-positive-Helicobacter pylori infection. AB - AIM: To elucidate the sequential gene expression profile in AGS cells co-cultured with wild-type Helicobacter pylori (H pylori) as a model of H pylori-infected gastric epithelium, and to further examine the contribution of cag-pathogenicity islands (cagPAI)-coding type IV secretion system and the two pathways, nuclear factor kappa B (NF-kappaB) and extracellular signal-regulated kinases (ERK) on wild-type H pylori-induced gene expression. METHODS: Gene expression profiles induced by H pylori were evaluated in AGS gastric epithelial cells using cDNA microarray, which were present in the 4 600 independent clones picked up from the human gastric tissue. We also analyzed the contribution of NF-kappaB and ERK signaling on H pylori-induced gene expression by using inhibitors of specific signal pathways. The isogenic mutant with disrupted cagE (Delta cagE) was used to elucidate the role of cagPAI-encoding type IV secretion system in the gene expression profile. RESULTS: According to the expression profile, the genes were classified into four clusters. Among them, the clusters characterized by continuous upregulation were most conspicuous, and it contained many signal transducer activity-associated genes. The role of cagPAI on cultured cells was also investigated using isogenic mutant cagE, which carries non-functional cagPAI. Then the upregulation of more than 80% of the induced genes (476/566) was found to depend on cagPAI. Signal transducer pathway through NF-kappaB or ERK are the major pathways which are known to be activated by cagPAI-positive H pylori. The role of these pathways in the whole signal activation by cagPAI-positive H pylori was analyzed. The specific inhibitors against NF-kappaB or ERK pathway blocked the activation of gene expression in 65% (367/566) or 76% (429/566) of the genes whose activation appealed to depend on cagPAI. CONCLUSION: These results suggest that more than half of the genes induced by cagPAI-positive H pylori depend on NF-kappaB and ERK signaling activation, and these pathways may play a role in the gene expression induced by host-bacterial interaction which may associate with H pylori-related gastro-duodenal diseases. PMID- 16273642 TI - Identification of anrF gene, a homology of admM of andrimid biosynthetic gene cluster related to the antagonistic activity of Enterobacter cloacae B8. AB - AIM: To identify the gene (s) related to the antagonistic activity of Enterobacter cloacae B8 and to elucidate its antagonistic mechanism. METHODS: Transposon-mediated mutagenesis and tagging method and cassette PCR-based chromosomal walking method were adopted to isolate the mutant strain(s) of B8 that lost the antagonistic activity and to clone DNA fragments around Tn5 insertion site. Sequence compiling and open reading frame (ORF) finding were done with DNAStar program and homologous sequence and conserved domain searches were performed with BlastN or BlastP programs at www.ncbi.nlm.nih.gov. To verify the gene involved in the antagonistic activity, complementation of a full-length clone of the anrF gene to the mutant B8F strain was used. RESULTS: A 3 321 bp contig around the Tn5 insertion site was obtained and an ORF of 2 634 bp in length designated as anrF gene encoding for a 877 aa polyketide synthase-like protein was identified. It had a homology of 83% at the nucleotide level and 79% ID/87% SIM at the protein level, to the admM gene of Pantoea agglomerans andrimid biosynthetic gene cluster (AY192157). The Tn5 was inserted at 2 420 bp of the gene corresponding to the COG3319 (the thioesterase domain of type I polyketide synthase) coding region on B8F. The antagonistic activity against Xanthomonas oryzae pv. oryzae was resumed with complementation of the full-length anrF gene to the mutant B8F. CONCLUSION: The anrF gene obtained is related to the antagonistic activity of B8, and the antagonistic substances produced by B8 are andrimid and/or its analogs. PMID- 16273643 TI - Receptor-binding domain of SARS-Cov spike protein: soluble expression in E. coli, purification and functional characterization. AB - AIM: Spike protein of coronavirus is responsible for virus binding, fusion and entry, and is a major inducer of neutralizing antibodies. This paper was to find a soluble and functional recombinant receptor-binding domain of severe acute respiratory syndrome-associated coronavirus (SARS-Cov), and to analyze its receptor binding ability. METHODS: Three fusion tags (glutathione S-transferase, GST; thioredoxin, Trx; maltose-binding protein, MBP), which preferably contributes to increasing solubility and to facilitating the proper folding of heteroprotein, were used to acquire the soluble and functional expression of RBD protein in Escherichia coli (BL21(DE3) and Rosetta-gamiB(DE3) strains). The receptor binding ability of the purified soluble RBD protein was then detected by ELISA and flow cytometry assay. RESULTS: RBD of SARS-Cov spike protein was expressed as inclusion body when fused as TrxA tag form in both BL21 (DE3) and Rosetta-gamiB (DE3) under many different cultures and induction conditions. And there was no visible expression band on SDS-PAGE when RBD was expressed as MBP tagged form. Only GST tagged RBD was soluble expressed in BL21(DE3), and the protein was purified by AKTA Prime Chromatography system. The ELISA data showed that GST/RBD antigen had positive reaction with anti-RBD mouse monoclonal antibody 1A5. Further flow cytometry assay demonstrated the high efficiency of RBD's binding ability to ACE2 (angiotensin-converting enzyme 2) positive Vero E6 cell. And ACE2 was proved as a cellular receptor that meditated an initial affinity interaction with SARS-Cov spike protein. The geometrical mean of GST and GST/RBD binding to Vero E6 cells were 77.08 and 352.73 respectively. CONCLUSION: In this paper, we get sufficient soluble N terminal GST tagged RBD protein expressed in E.coli BL21(DE3); data from ELISA and flow cytometry assay demonstrate that the recombinant protein is functional and binding to ACE2 positive Vero E6 cell efficiently. And the recombinant RBD derived from E.coli can be used to developing subunit vaccine to block S protein binding with receptor and to neutralizing SARS-Cov infection. PMID- 16273644 TI - Effects of Saccharomyces boulardii on fecal short-chain fatty acids and microflora in patients on long-term total enteral nutrition. AB - AIM: Diarrhea is a significant problem in patients on total enteral nutrition (TEN) and may involve changes in intestinal short chain fatty acids or microflora. Recent studies suggest that the probiotic yeast Saccharomyces boulardii (Sb) may decrease its incidence. The aim of this study was to assess the effects of Sb on fecal flora and short-chain fatty acids (SCFA) in patients on long-term TEN. METHODS: Ten patients (3 females, 7 males, 59+/-5.5 years), on TEN for a median of 13 mo (1-125), and 15 healthy volunteers (4 females, 11 males, 32+/-2.0 years) received Sb (0.5 g bid PO) for 6 d. Two stool samples were taken before, on the last 2 d and 9-10 d after treatment, for SCFA measurement and for culture and bacterial identification. Values (mean+/-SE) were compared using sign tests and ANOVA. RESULTS: Fecal butyrate levels were lower in patients (10.1+/-2.9 mmol/kg) than in controls (19.2+/-3.9, P = 0.02). Treatment with Sb increased total fecal SCFA levels in patients (150.2+/-27.2 vs 107.5+/-18.2 mmol/kg, P = 0.02) but not in controls (129.0+/-28.6 vs 113.0+/-15.2 mmol/kg, NS). At the end of treatment with Sb, patients had higher fecal butyrate (16.0+/ 4.4 vs 10.1 [2.9] mmol/kg, P = 0.004). Total SCFAs remained high 9 d after treatment was discontinued. Before the treatment, the anaerobe to aerobe ratio was lower in patients compared to controls (2.4+/-2.3 vs 69.8+/-1.8, P = 0.003). There were no significant changes in the fecal flora of TEN patients. CONCLUSION: Sb-induced increase of fecal SCFA concentrations (especially butyrate) may explain the preventive effects of this yeast on TEN-induced diarrhea. PMID- 16273645 TI - Prevalence of cholelithiasis in patients with chronic inflammatory bowel disease. AB - AIM: To investigate the effect of chronic inflammatory bowel disease (CIBD) specific risk factors for cholecystolithiasis, as duration and involvement pattern of the disease and prior surgery in patients with Crohn's disease (CD) and ulcerative colitis (UC). METHODS: A total of 222 patients with CD (135 females, 87 males; average age, 35.8+/-11.8 years; range 17-81 years) and 88 patients with UC (39 females, 49 males; average age, 37.2+/-13.6 years; range 16 81 years) underwent clinical and ultrasound examinations. Besides age, sex and degree of obesity, patients' CIBD specific parameters, including duration and extent of disease and prior operations were documented and evaluated statistically using logistic regression. RESULTS: The overall prevalence of gallbladder stone disease in patients with CD was 13% (n = 30). Only age could be shown to be an independent risk factor (P = 0.014). Compared to a collective representative for the general population in the same geographic region, the prevalence of cholecystolithiasis was higher in all corresponding age groups. Patients with UC showed an overall prevalence of gallbladder stone disease of only 4.6%. CONCLUSION: Disease-specific factors such as duration and extent of disease, and prior surgery are independent risk factors for the development of cholecystolithiasis in patients with CIBD. PMID- 16273646 TI - Quantitative evaluation of long-term liver repopulation and the reconstitution of bile ductules after hepatocellular transplantation. AB - AIM: The treatment of liver disease is severely limited by a shortage of donor livers. In trying to address this growing problem, hepatocellular transplantation (HTx) has received much attention as an alternative to whole organ transplant. However, the expansion of transplanted cells is at low level, and the reconstitution of functional liver tissue is limited by this cellular property. We set up an animal model to better understand cell dose effect and the kinetics of liver repopulation following HTx. METHODS: Dipeptidyl peptidase IV (DPPIV) deficient rats treated with retrorsine and subjected to partial hepatectomy were infused with DPPIV-positive hepatocytes. Rats were injected with varying numbers of donor hepatocytes down to 100 cells low, and liver repopulation was examined at different time points up to 20 mo long. Repopulation was assessed by computer aided quantitative detection. RESULTS: Transplanted hepatocytes underwent multiple rounds of proliferation and stably repopulated the injured livers after 20 mo and at all cell doses. Transplanted cells divided 14 times within the 3-mo time period following infusion, and the liver repopulation reached a plateau between 3 and 20 mo. Approximately 90% replacement occurred. Donor-derived cells also reconstituted the bile ductules of the recipients. CONCLUSION: The ability of transplanted hepatocytes to fully reconstitute injured livers strongly supports further investigation into the clinical potential of HTx. Additionally, the observation that transplanted hepatocytes also form components of the biliary system suggests that these cells may have bi-potential property of the stem cells. PMID- 16273647 TI - Serum leptin levels and insulin resistance are associated with gallstone disease in overweight subjects. AB - AIM: To establish an association between the serum leptin levels and the development of gallstone disease (GD). METHODS: We carried out a non-matched case controlled study in a university hospital in Mexico City. Two hundred and eighty seven subjects were included: 97 cases with gallstones and 190 controls. Body mass index (BMI), fasting plasma leptin, insulin, serum lipid, and lipoprotein levels were measured. Insulin resistance was calculated by homeostasis model assessment (HOMA-IR). Unconditional logistic regression analysis (univariate and multivariate) stratified by BMI was used to calculate the risk of GD. RESULTS: The multivariate conditional regression analysis revealed a model for those patients with BMI <30. The selected variables in the model were HOMA-IR index with OR = 1.31, P = 0.02 and leptin higher than median with OR = 2.11, P = 0.05. In the stratum of BMI >=30, we did not find a useful model. CONCLUSION: We concluded that insulin resistance and the development of GD appears to be associated with serum leptin levels in subjects with overweight, but not in obese subjects with similar metabolic profiles. PMID- 16273648 TI - Treatment of genotype 2 and 3 chronic hepatitis C virus-infected patients. AB - AIM: Before pegylated interferon alpha (IFN) was introduced for the therapy of chronic hepatitis C virus (HCV)-induced hepatitis, conventional thrice weekly IFN therapy was supplemented by ribavirin. Also, at that time, higher and more frequent doses of IFN were expected to be more effective than the standard regimen of 3 MU thrice weekly. As ribavirin significantly increases side effects and negatively influences the quality of life particularly in young patients, we started a prospective non-randomized study with a daily IFN-2a monotherapy as an initial treatment for chronic hepatitis C. METHODS: Forty-six consecutive chronic HCV-infected patients received 3 MU IFN-2a per day as an initial treatment. Patients with genotype 2 or 3 (n = 12) were treated for 24 wk, and patients with genotypes other than 2 or 3 (n = 34) for 48 wk. Treatment outcome was followed up for 48 wk after the end of treatment (EOT). Virological response was defined as the absence of detectable serum HCV-RNA. Patients without virological response at 12 wk after the start of treatment received low-dose ribavirin (10 mg(kg/d)) additionally. RESULTS: During treatment, three genotype 3 patients were excluded from the study due to incompliance. The remaining patients (n = 9) infected with genotype 2 or 3 showed an initial virological response rate of 100%. Six patients (66.7%) were still found to be virus-free at the end of follow-up period. In these patients, initial virological response was evident already after 2 wk of treatment. In contrast, initial virological response occurred first after 4 wk of treatment in the three patients who relapsed (33.3%). In comparison, patients infected with genotypes other than 2 or 3 (n = 34) showed an initial virological response rate of only 23.5% (n = 8), and even in combination with ribavirin a sustained virological response (SVR) rate of only 11.8% (n = 4) could be achieved. CONCLUSION: In chronic HCV-infected patients with genotype 2 or 3, a SVR can be expected after 24 wk of daily dose IFN-2a treatment without ribavirin, if initial virological response develops early. This finding is worth to be confirmed in a prospective randomized study with pegylated IFN. PMID- 16273649 TI - Quantitative analysis of plasma HBV DNA for early evaluation of the response to transcatheter arterial embolization for HBV-related hepatocellular carcinoma. AB - AIM: Transcatheter arterial embolization (TAE) is an important palliative treatment for patients with hepatocellular carcinoma (HCC) who are poor candidates for surgery or percutaneous ablative therapy. It generally takes 4 wk after lipiodol-TAE to properly assess lipiodol retention on computed tomography (CT). HBV DNA is integrated into the genome of HCC cells, and circulating plasma DNA may serve as a marker for cell damage. We assessed changes in plasma HBV DNA after TAE in HBV-related HCC and correlated the levels with the pattern of lipiodol accumulation on CT. METHODS: Between April and June 2001, 14 patients with HBV-associated HCC who underwent TAE for inoperable or recurrent tumor were studied. Levels of plasma HBV DNA were measured by real-time quantitative PCR daily for five consecutive days after TAE. More than twofold elevation of circulating HBV DNA was considered as a definite elevation. Abdominal CT was performed 1-2 mo after TAE for the measurement of lipiodol retention. RESULTS: Circulating HBV DNA in 10 out of 13 patients was elevated after TAE, except for one patient whose plasma HBV DNA was undetectable before and after TAE. In group I patients (n = 6), the HBV DNA elevation persisted for more than 2 d, while in group II (n = 7), the HBV DNA elevation only appeared for 1 d or did not reach a definite elevation. There were no significant differences in age or tumor size between the two groups. Patients in group I had significantly better lipiodol retention (79.31+/-28.79%) on subsequent abdominal CT than group II (18.43+/ 10.61%) (P = 0.02). CONCLUSION: Patients with durable HBV DNA elevation for more than 2 d correlated with good lipiodol retention measured 1 mo later, while others associated with poor lipiodol retention. Thus, circulating HBV DNA may be an early indicator of the success or failure of TAE. PMID- 16273650 TI - Glutamine-supplemented total parenteral nutrition attenuates plasma interleukin-6 in surgical patients with lower disease severity. AB - AIM: Previous reports have shown that decrease in plasma glutamine (Gln) level following major surgery may contribute to the state of immunosuppression. Gln supplementation improves the depletion of body Gln pool, and may have indirect effect on reducing proinflammatory mediator release. This study evaluated whether the effect of Gln dipeptide-enriched total parenteral nutrition (TPN) on postoperative cytokine alteration depended on the disease severity of surgical patients. METHODS: Forty-eight patients with major abdominal surgery were allocated to two groups to receive isonitrogenous (0.228 g nitrogen/kg per d) and isocaloric (30 kcal/kg per d) TPN for 6 d. Control group (Conv) using conventional TPN solution received 1.5 g amino acids/kg per day, whereas the test group received 0.972 g amino acids/kg per day and 0.417 g L-alanyl-L-glutamine (Ala-Gln)/kg per day. Blood samples were collected on d 1 and d 6 postoperatively for plasma interleukin (IL)-2, IL-6, IL-8, and interferon (IFN)-gamma analysis. RESULTS: Plasma IL-2 and IFN-gamma were not detectable. IL-6 concentrations were significantly lower on the 6(th) postoperative day in the Ala-Gln group than those in the Conv group in patients with APACHE II <=6, whereas no difference was noted in patients with APACHE II >6. There was no difference in IL-8 levels between the two groups. No difference in cumulative nitrogen balance was observed on d 2-5 after the operation between the two groups (Ala-Gln -3.2+/-1.6 g vs Conv -6.5+/-2.7 g). A significant inverse correlation was noted between plasma IL-6 levels and cumulative nitrogen balance postoperatively in the Ala-Gln group, whereas no such correlation was observed in the Conv group. CONCLUSION: TPN supplemented with Gln dipeptide had no effect on plasma IL-8 levels after surgery. However, Gln supplementation had a beneficial effect on decreasing systemic IL-6 production after surgery in patients with low admission illness severity, and lower plasma IL-6 may improve nitrogen balance in patients with abdominal surgery when Gln was administered. PMID- 16273651 TI - Expression of serine protease SNC19/matriptase and its inhibitor hepatocyte growth factor activator inhibitor type 1 in normal and malignant tissues of gastrointestinal tract. AB - AIM: To provide the expression profile of serine protease SNC19/matriptase and its inhibitor hepatocyte growth factor activator inhibitor type 1 (HAI-1) in normal and malignant tissues of gastrointestinal tract at mRNA level for further study on their correlations with tumor progression and metastasis. METHODS: Total RNAs were prepared from 37 samples of colorectal cancer tissues, 40 samples of gastric cancer tissues, and their adjacent normal tissues. The expression of SNC19/matriptase and HAI-1 in these samples was detected by real-time fluorescent quantitative PCR using glyceraldehyde-3-phosphate dehydrogenase as internal standard, and the clinical significance for the correlation with clinicopathological parameters was evaluated. RESULTS: In gastric cancer tissues the expression of HAI-1 and SNC19/matriptase was significantly lower than that in the corresponding adjacent normal tissues (Z = -3.280, P = 0.006; Z = -4.651, P = 0.000). HAI-1:SNC19/matriptase ratio showed no difference between normal and malignant tissues (P>0.05). Analysis of clinicopathological parameters showed decreased expression of HAI-1 and HAI-1:SNC19/matriptase ratio associated with stage III/IV gastric tumors as compared to stage I/II ones (Z = -2.140, P = 0.031; Z = -2.155, P = 0.031), and with lymph node-positive gastric cancer tissues as compared to lymph node-negative ones (Z = -2.081, P = 0.036; Z = 2.686, P = 0.006). The expression of SNC19/matriptase had no relationship with stages and lymph node metastasis (P>0.05). The expression of HAI-1 and HAI 1:SNC19/matriptase ratio increased in well-differentiated gastric cancer tissues, but there was no statistical significance (P>0.05). The difference of SNC19/matriptase expression was not significant in gastric cancer tissues of different histological differentiation status (P>0.05). In colorectal cancer tissues, the expression of HAI-1 and SNC19/matriptase was also markedly lower than that in their adjacent normal tissues (Z = -3.100, P = 0.002; Z = -2.731, P = 0.006), whereas HAI-1:SNC19/matriptase ratio showed no difference. Decreased expression of HAI-1 was associated with increased invasive depth and lymph node metastasis, but there was no statistical significance (P>0.05). The difference of SNC19/matriptase expression and HAI-1:SNC19/matriptase ratio was not significant in different stages and different lymph node metastasis status (P>0.05). The expression of SNC19/matriptase, HAI-1 or HAI-1:SNC19/martiptase ratio showed no difference in colorectal cancer tissues of different histological differentiation status (P>0.05). CONCLUSION: The expressions of SNC19/matriptase and its inhibitor HAI-1 are decreased in gastrointestinal cancer tissues compared to their normal counterparts, and the decreased expression of HAI-1 may correlate with invasion and lymph node metastasis. The possible mechanisms involved need to be further investigated. PMID- 16273652 TI - Microcirculation disturbance affects rats with acute severe pancreatitis following lung injury. AB - AIM: To study the effects of microcirculation disturbance (MD) on rats with acute severe pancreatitis (ASP). METHODS: We developed ASP rat models, and anatomized separately after 1, 3, 5, 7, and 9 h. We took out blood and did hemorrheologic examination and erythrocyte osmotic fragility test, checked up the water content, capillary permeability, and genetic expression of intercellular adhesion molecule 1 (ICAM-1) in lung tissues, examined the apoptosis degree of blood vessel endothelium while we tested related gene expression of Bax and Bcl-2 in lung tissues. We did the same examination in control group. RESULTS: The viscosity of total blood and plasma, the hematocrit, and the erythrocyte osmotic fragility were all increased. Fibrinogen was decreased. The water content in lung tissues and capillary permeability were increased. Apoptosis degree of blood vessel endothelium was increased too. ICAM-1 genetic expression moved up after 1 h and reached its peak value after 9 h. CONCLUSION: MD plays an important role in ASP following acute lung injury (ALI). The functional damage of blood vessel endothelium, the apoptosis of capillary vessel endothelium, WBC edging concentration and the increasing of erythrocyte fragility are the main reasons of ALI. PMID- 16273653 TI - Effects of hepatitis B virus on p53 expression in hepatoma cell line SMMU-7721. AB - AIM: To investigate the contribution of HBV in the development of hepatocarcinoma by examining the effects of HBV on p53 function in SMMU-7721 cell line. METHODS: Plasmid pCMVp53 was transfected or cotransfected with pCMVHBVa (wild-type HBV) or PCMVHBVb (mutation type HBV) into the hepatoma cell line SMMU-7721 by lipofectamine. Apoptosis cells were labeled with annexin V-FITC and confirmed by flow cytometry. Reporter plasmid PG13-CAT or p21-luc was cotransfected, respectively, into each group to determine the transactivation activity of p53 and its effect on p21 promoter. Western blot was performed to observe p53 expression in hepatoma cell line of each group. RESULTS: The group transfected with pCMVp53 alone exhibited higher luciferase activity and higher apoptosis rate, otherwise, the p53 expression and reporter activity of PG13-CAT or P21-luc as well as cell apoptosis rate were obviously higher in the group cotransfected of pCMVp53 with pCMVHBVa, but not in the other cotransfected group. CONCLUSION: Transient transfection of HBV into the SMMU-7721 cell line can enhance p53 expression and its effects on development of hepatocarcinoma. PMID- 16273654 TI - Alterations of intestinal immune function and regulatory effects of L-arginine in experimental severe acute pancreatitis rats. AB - AIM: To discuss the changes of intestinal mucosal immune function in rats with experimental severe acute pancreatitis (SAP) and the regulatory effect of L arginine. METHODS: Male adult Wistar rats were randomly divided into pancreatitis group, sham-operation group, and L-arginine treatment group. Animals were killed at 24, 48, and 72 h after SAP models were developed and specimens were harvested. Endotoxin concentration in portal vein was determined by limulus endotoxin analysis kit. CD3+, CD4+, CD8+ T lymphocytes in intestinal mucosal lamina propria were examined by immunohistochemistry. Secretory immunoglobulin A (SIgA) in cecum feces was examined by radioimmunoassay. RESULTS: Compared to the control group, plasma endotoxin concentration in the portal vein increased, percentage of CD3+ and CD4+ T lymphocyte subsets in the end of intestinal mucosal lamina propria reduced significantly, CD4+/CD8+ ratio decreased, and SIgA concentrations in cecum feces reduced at 24, 48, and 72 h after SAP developed. Compared to SAP group, the L-arginine treatment group had a lower level of plasma endotoxin concentration in the portal vein, a higher CD3+ and CD4+ T lymphocyte percentage in the end of intestinal mucosal lamina propria, an increased ratio of CD4+/CD8+ and a higher SIgA concentration in cecum feces. CONCLUSION: Intestinal immune suppression occurs in the early stage of SAP rats, which may be the main reason for bacterial and endotoxin translocation. L-arginine can improve the intestinal immunity and reduce bacterial and endotoxin translocation in SAP rats. PMID- 16273655 TI - Primary biliary cirrhosis after aortoiliac reconstruction surgery using a Y graft. AB - Primary biliary cirrhosis (PBC) is an autoimmune disease characterized by anti mitochondrial antibodies and destruction of intra-hepatic bile ducts. Though little is known about the etiology of PBC, some reports suggest that xenobiotics and viral infections may induce PBC. We describe a case of PBC after the aortoiliac reconstruction surgery using a Y-graft. PMID- 16273656 TI - Primary pancreatic anaplastic large cell lymphoma, ALK negative: a case report. AB - We present the fourth case of a primary pancreatic anaplastic large cell lymphoma (ALCL), ALK-. An 80-year-old man was admitted to our clinic for further investigation of a fever of unknown origin. He noted anorexia, weight loss and fatigue. His laboratory tests showed anemia and a great elevation of ESR, LDH, and beta (2) microglobulin. In CT and MRI scan, a soft tissue mass in the pancreas was observed. A repeated endoscopy after his admission revealed an ulcerated mass-like deformity of the duodenal bulb. Explorative laparotomy confirmed a diffuse spread of an unresectable malignant pancreatic mass extending to the adjacent organs. Duodenal and surgical biopsies identified an ALCL of T cell lineage, ALK-. The patient died in the Intensive Care Unit due to hemodynamic instability. Our case is the first one indicating that primary pancreatic lymphoma should be suspected in a patient with pancreatic mass and elevated serum LDH and beta(2) microglobulin. PMID- 16273657 TI - Adenosquamous carcinoma arising within a retrorectal tailgut cyst: report of a case. AB - Retrorectal, developmental tail gut cysts, include dermoid cysts, rectal duplication cysts and retrorectal cyst-hamartomas. Retrorectal cyst-hamartomas (RCH) are derived from remnants of the tail gut, the most caudal part of the embryonic hind gut, which normally involutes by the 8(th) wk of embryonic development (3-8 mm stage). They have specific radiological and histopathological features that distinguish them from other similar formations (dermoid cysts, enteric duplication cysts and teratomas). We report a patient with adenosquamous carcinoma arising within RCH, who underwent complete resection of the cyst through anterior laparotomy, and reached complete (recurrence-free for 14 mo, so far) functional recovery. The cyst was incidentally discovered during hysterectomy 12 years ago. Diagnostic, therapeutic and histopathological aspects of this rare case are discussed. The mentioned period between diagnosis and surgical treatment suggests that RCH, given enough time, can develop malignant degeneration, and should be resected at the time of diagnosis. PMID- 16273658 TI - Agenesis of the gallbladder: a dangerously misdiagnosed malformation. AB - Isolated agenesis of the gallbladder is a rare anomaly, often asymptomatic. However, one patient out of four presented with right upper abdominal pain, nausea, and fatty food intolerance. The condition is frequently mistaken with an excluded or sclero-atrophic gallbladder, regardless of the imaging modality used. Consequently, AG often leads to unnecessary and potentially dangerous laparoscopic surgery as described in a few case reports over the last 10 years. The aim of this study is to clarify the diagnostic and therapeutic approach of this unusual pathology. Two cases seen in our institutions were retrospectively reviewed, together with a review of the American and European literature. During laparoscopy, the absence of normal anatomical structures and the impossibility of pulling on the gallbladder to expose and dissect the triangle of Callot increases the risk of iatrogenic injury to biliary or portal structures. Depending on the experiment of the surgeon in laparoscopic procedure, this has to be taken into account to decide a conversion to laparotomy. A high index of suspicion is necessary when interpreting the radiological images. In case of doubt, a MRI cholangiography is mandatory. Because of possible inherited transmission, relatives with a history of biliary symptoms should be investigated. PMID- 16273659 TI - Treatment of a duodenal perforation secondary to an endoscopic sphincterotomy with clips. AB - Perforation is one of the most serious complications of endoscopic sphincterotomy (ES) necessitating immediate surgical intervention. We present a case of successful management of such a complication with endoclipping. A 85-year-old woman developed duodenal perforation after ES. The perforation was identified early and its closure was achieved using three metallic clips in a single session. There was no procedure-related morbidity or complications and our patient was discharged from hospital 10 d later. Endoclipping of duodenal perforation induced by ES is a safe, effective and alternative to surgery treatment. PMID- 16273661 TI - Abstracts of the XXIII World Congress of Pathology and Laboratory Medicine (WASPaLM), Istanbul, Turkey, May 26-30, 2005. PMID- 16273660 TI - Ruptured hepatoblastoma with massive internal bleeding in an adult. AB - Hepatoblastoma is the most common primary hepatic tumor of children. However, only a very few cases have been reported in adults. Most studies support treatment with chemotherapy followed by surgical resection. We present the first reported case of adult hepatoblastoma in Taiwan. A 52-year-old female suffered from sudden onset of abdominal pain and general weakness for days. Internal bleeding with hemorrhagic shock was suspected and two massive lesions in both lobes of the liver with hemoperitoneum were noted from imaging studies. Surgical resection of the larger left lobe tumor and radio-frequency ablation of the right smaller one were performed. The histopathology diagnosis was of a hepatoblastoma. PMID- 16273662 TI - A soldier with traumatic brain injury and meningitis. PMID- 16273663 TI - Creation of new national committee on bioethics. PMID- 16273664 TI - Food, glorious food? PMID- 16273665 TI - Fungi behaving badly. PMID- 16273666 TI - Prediction of mesorectal nodal metastases after chemoradiation for rectal cancer: results of a randomised trial: implication for subsequent local excision. AB - BACKGROUND AND PURPOSE: For patients with rectal cancer treated with full thickness local excision the risk of mesorectal nodal metastases has to be very low. The aim was to assess this risk after preoperative radiotherapy in relation to pathological T-category. PATIENTS AND METHODS: Three hundred sixteen patients with resectable cT3-4 low rectal carcinoma were randomised to receive either pre operative 5 x 5 Gy irradiation with subsequent surgery performed within 7 days or chemoradiation (50.4, 1.8 Gy per fraction plus bolus 5-fluorouracil and leucovorin) followed by surgery after 4-6 weeks. The pathological reports of patients who fulfilled entry criteria and had preoperative irradiation followed by transabdominal surgery were analysed. RESULTS: Significant downstaging of primary tumour (P<0.001) and of nodal disease (P=0.007) was observed after chemoradiation in comparison with short-course irradiation. In chemoradiation group, for patients with complete pathological response and for ypT1 category, the rate of nodal metastases was low - 5% (95% confidence interval [CI] 0-14%) and 8% (95% CI 0-24%), respectively. The rate of ypN-positive disease in chemoradiation group was similar to that recorded in short-course irradiation group for ypT2 category 26% (95% CI 14-38%) vs. 28% (95% CI 16-40%), P=0.83 and for ypT3-4 category 55% (95% CI 41-69%) vs. 64% (95% CI 54-74%), respectively, P=0.37. For ypT2 category after chemoradiation, the rate of nodal disease remained high even in subgroup with low residual cancer cells density (20%, 95% CI 4-36%). CONCLUSIONS: For patients with tumours downstaged by chemoradiation to ypT0 and ypT1 full thickness local excision may be considered as an acceptable approach, because the risk of mesorectal lymph nodes metastases is low. The selection criteria for preoperative radio(chemo)therapy and local excision are discussed. PMID- 16273667 TI - Wilhelm Fresenius Memorial Issue. PMID- 16273668 TI - Professor David F. Horrobin 1939-2003: A tribute. Proceeding from the 1st Brain Phospholipids Conference, Aviemore, Scotland, United Kingdom, August/September 2003. PMID- 16273669 TI - Endocrine-related resources from the National Institutes of Health. PMID- 16273670 TI - F.D.A. to weigh at-home testing for AIDS virus. PMID- 16273671 TI - Appendix: acute exposure guideline levels (AEGLs) toxicity and properties of selected chemical and biological agents. PMID- 16273672 TI - Fetal interests vs. maternal rights: is the state going too far? PMID- 16273673 TI - Delicate balance. PMID- 16273674 TI - [Are most of the alternative procedures ineffective? Test of substances in complementary medicine (interview by Dr. Thomas Meissner)]. PMID- 16273676 TI - Continuous intravenous heparin infusion prevents peri-operative thromboembolic events in bariatric surgery patients. AB - BACKGROUND: The pharmacokinetics of subcutaneous heparin administration in the obese patient are unpredictable. Peak levels are slowly reached and the effects are not rapidly reversible. Low-dose, continuous, intravenous heparin is easily reversed, is more efficacious and is cost-effective. METHODS: From November 2000 until July 2005, 822 consecutive patients were administered continuous intravenous unfractionated heparin at 400 U/hr (9,600 U/day) starting in the preoperative holding area and maintained until discharge. All clinically significant events were documented. RESULTS: 634 laparoscopic gastric bypass, 10 revisions and 188 Lap-Band procedures were performed. The mean age was 43+/-11 years (15-74) and mean BMI was 45.2+/-7.1 (30-86). There was only one (0.12%) clinically evident thromboembolic event in the entire cohort (after a gastric bypass). Anti-Xa levels and prothrombin time were followed in a group of 40 patients and were found to be normal in all. Bleeding that required transfusion occurred in 1.3% of patients. In 41 patients (5%), heparin therapy was terminated or temporarily held due to need for extensive adhesiolysis or acute drop in hematocrit, with-or-without other evidence of postoperative bleeding. Average estimated blood loss during surgery was 36 cc (5-500 cc). One patient was inadvertently administered excessive doses of heparin due to a pump error without significant sequelae. CONCLUSIONS: Continuous low-dose intravenous heparin therapy is associated with an extremely low incidence of thromboembolic events and a low risk for perioperative hemorrhage. Intravenous heparin also has the benefits of being inexpensive and rapidly reversible. PMID- 16273675 TI - An analysis of response-blocking parameters in the prevention of pica. AB - We manipulated two parameters of response blocking to reduce pica: (a) the criteria for initiating the procedure (either earlier or later in the response chain) and (b) the distance from which the procedure was initiated. Results suggested that response blocking may be effective only when implemented early in the chain and with near-perfect consistency. Further, additional treatment components may be required to eliminate all pica attempts. PMID- 16273677 TI - The disjunction between policy and practice: HIV discrimination in health care and employment in Indonesia. AB - Through the areas of health and employment, this study examined the disjunction between policy and practice with regards to HIV/AIDS-related discrimination in Indonesia. The primary centres for data collection were Jakarta and Bali, where it was found that the contradictions between international commitments and written internal policies supported for forms of arbitrary discrimination against People Living with HIV/AIDS (PLWHA). This paper argues that for effective change to occur, the cultural mindset of service providers in the health care and employment sectors need to be changed through integrating institutional support and educational training. PMID- 16273678 TI - Seeking a better solution for the disposition of frozen embryos: is embryo adoption the answer? PMID- 16273679 TI - An Oklahoma perspective: end of life decision-making and termination of treatment. PMID- 16273680 TI - Prescription for fairness: health insurance reimbursement for Viagra and contraceptives. PMID- 16273681 TI - Autonomy and death. AB - In this Article, Professor Clark explores the contours of the current debate over physician-assisted death. She beings by focusing on the legal issues raised by statutory attempts to either legalize or criminalize physician-assisted death, with particular emphasis on the constitutional questions that are currently before the United States Supreme Court. She then examines physician-assisted death from both medical and societal perspectives. Professor Clark uses a thought experiment in which assisted death is facilitated by persons other than physicians, and in doing so, questions whether physicians are the proper persons in whom to rest power over assisted death. She points out the irony in a process that would set up physicians as protectors of individual autonomy, and ultimately concludes that by deferring to the medical profession in this process, we risk losing the very autonomy that assisted death is designed to effectuate. PMID- 16273682 TI - Autonomy, justice, and disability. AB - In this Article, Professor Carlos A. Ball explores the philosophical foundations for the types of rights and benefits that our society currently provides to individuals with disabilities. The concept of autonomy places on society a moral obligation to assist individuals with disabilities when their basic human functional capabilities are impaired. The exercise of this obligation entails assisting individuals with crossing a minimum threshold of functional capabilities below which it is not possible to lead autonomous lives. In making this argument, Professor Ball responds to libertarian critics who contend that notions of freedom or liberty proscribe an activist role for government in this arena. He explains how even a libertarian state redistributes wealth in order to provide for some incapacities. Professor Ball also disputes the idea that the meeting of the needs of the disabled is enough to provide moral justification for the rights and benefits provided to individuals with disabilities. The problem with the concept of needs, Professor Ball argues, is that it fails to account sufficiently for the human good of personal autonomy. PMID- 16273683 TI - Pulmonary arterial hypertension. PMID- 16273684 TI - Lochner redeemed: family privacy after Troxel and Carhart. AB - At least since its 1992 decision in Planned Parenthood of Southeastern Pennsylvania v. Casey, the Supreme Court has differentiated its review of abortion laws from its scrutiny of other intrusions on family privacy. Whereas abortion restrictions are reviewed under the middling "undue burden" standard, incursions on other family-related liberties, including marriage, kinship, and child rearing are said to be subject to the strict scrutiny ordinarily employed in the defense of fundamental rights. This Article contends that the Court's most recent decisions in this context give reason to reconsider both sides of that equation. Stenberg v. Carhart, striking down Nebraska's ban on "partial-birth" abortions, suggests that the Court's scrutiny in the abortion context will be more aggressive and rigid than most had supposed. At the same time, its decision in Troxel v. Granville, limiting states' authority to order grandparent visitation over the objects of a parent, suggest that there is more fluidity in the Court's review of other family liberties than is conventionally assumed. Together the cases signal a convergence in both sorts of family-privacy controversies toward a common standard of "reasonableness." That standard bears, for many, an uncomfortable association with the much-maligned "natural law-due process formula" of the Lochner era, but Professor Meyer argues that it is precisely the right approach in the context of family privacy. Although more rigid doctrinal formulae are sometimes preferred on the ground that they constrain judicial judgment, here they are undesirable precisely because they obscure the value judgments that are inevitably at the core of every family privacy decision. PMID- 16273685 TI - Characterization of memory profile in subjects with amnestic mild cognitive impairment. AB - Different aspects of episodic long-term, short-term and implicit long-term memory were investigated in subjects who strictly fulfilled the criteria for the amnestic form of Mild Cognitive Impairment (a-MCI). Results showed normal short term memory abilities in these subjects, while each of the episodic long-term memory indices explored showed poorer results in a-MCI subjects with respect to normal controls. Although some episodic memory functions were relatively well preserved, others appeared to have deteriorated to a level comparable to that of mild AD patients. The finding of an extensive impairment of all memory functions depending on hippocampal structures in a population with a high risk of developing dementia is strongly supportive of the hypothesis that a pure amnesic syndrome characterizes the preclinical phase of AD. PMID- 16273686 TI - Why we need the independent sector: the behavior, law, and ethics of not-for profit hospitals. AB - Among the major forms of corporate ownership, the not-for-profit ownership form is distinct in its behavior, legal constraints, and moral obligations. A new empirical analysis of the American Hospital industry, using eleven years of data for all urban general hospitals in the country, shows that corporate form accounts for large differences in the provision of specific medical services. Not for-profit hospitals systematically provide both private and public goods that are in the public interest, and that other forms fail to provide. Two hypotheses are proposed to account for the findings, one legal and one moral. While no causal claims are made, not-for-profit hospital behavior is consistent with the behavior required by law and morality. The moral argument, developed as a preliminary theory of not-for-profit ethics, also provides a potential reason to prefer not-for-profit hospitals. The findings provide a new justification for the not-for-profit tax exemption for hospitals, and also suggest new uses for ownership categories as regulatory tools. PMID- 16273687 TI - Technology and the legal discourse of fetal autonomy. AB - The relationship between society, medicine, and the law is multi-faceted and complex. This Article examines the process of, and the influences on, the construction of fetal personhood in the legal discourses in American and Commonwealth case law and statutes. It demonstrates how the physical and visual separation of the fetus, as made possible by medical advances, has influenced the development of legal doctrine relating to the rights of the fetus. PMID- 16273688 TI - Increasing access to emergency contraceptive pills through state law enabled dependent pharmacist prescribers. AB - Emergency contraceptive pills ("ECPs"), a form of contraception which has a 75% chance of preventing pregnancy when taken within 72 hours after unprotected intercourse, are currently available only by prescription. Increasing access to ECPs will help to reduce unintended pregnancies and abortions and will help to provide women with an extra level of control over their reproductive futures. Private access initiatives, while helpful, are insufficient to address the access problem; federal level solutions are unlikely to be implemented soon. Thus, this Article proposes that all states adopt the Washington model for dependent pharmacist prescribers, whereby state law enables physicians to create collaborative agreements with pharmacists, pursuant to which the physician can effectively delegate to a pharmacist in a retail setting the power to prescribe ECPs. Dependent pharmacist prescribing of ECPs increases access to contraception, increases patient satisfaction and decreasing cost, while preserving patient safety through screening mechanisms and the good safety profile of ECPs. A state by-state analysis of pharmacy law reveals that while there is a national trend expanding the scope of pharmacy practice, states vary widely in the amount of prescribing power allowed to pharmacists. However, dependent pharmacist prescribing of ECPs is currently feasible in a few states and close in a number of others, and implementation of dependent pharmacist prescribing in some states may lead to greater nationwide acceptance of both dependent pharmacist prescribing and of ECPs. PMID- 16273689 TI - Don't rush to judgment on "Dolly": human cloning and its individual procreative liberty implications. PMID- 16273690 TI - Telemedicine: patient privacy rights of electronic medical records. PMID- 16273691 TI - When technology and health care collide: issues with electronic medical records and electronic mail. PMID- 16273692 TI - Ethical considerations in managed care: a commentary. PMID- 16273693 TI - Mainstreaming complementary and alternative medicine in the face of uncertainty. PMID- 16273694 TI - A backdoor to policy making: the use of philosophers by the Supreme Court. PMID- 16273695 TI - Better off dead than disabled?: should courts recognize a "wrongful living" cause of action when doctors fail to honor patients' advance directives? PMID- 16273696 TI - Financial conflicts of interest in human subjects research: proposals for a more effective regulatory scheme. PMID- 16273697 TI - AIDS and the blood supply: an analysis of law, regulation, and public policy. PMID- 16273698 TI - Sacrificing patients for profits: physician incentives to limit care and ERISA fiduciary duty. PMID- 16273699 TI - Surpassing the material: the human rights implications of informed consent in bioprospecting cells derived from indigenous people groups. PMID- 16273700 TI - Coercive abortions and criminalizing the birth of children: some thoughts on the impact on women of State v. Oakley. PMID- 16273701 TI - Ethical dilemmas in reproductive medicine. PMID- 16273702 TI - [Fibroangioma of the umbilical cord: report of one case]. PMID- 16273703 TI - The choice of participation by physicians in capital punishment. PMID- 16273704 TI - [Ovarian hyperstimulation syndrome associated with spontaneous pregnancy: report of two cases]. PMID- 16273705 TI - Bioethical and legal implications of pediatric gastric bypass. PMID- 16273706 TI - Immunohistological detection of growth factors and cytokines in tissue mast cells. AB - The relative rarity of mast cells (MCs) and the rich content of heparin in the cytoplasmic granules of MCs pose technical challenges in reliably detecting growth factors (GFs) or cytokines in MCs by conventional immunohistological stain (IHS) methods. A variety of polypeptide growth factors are characterized by high affinity to heparin. Binding of GFs to MC granules during detection can lead to highly specific yet falsely positive results that cannot be easily discovered by conventional procedure controls. Many reagents used in IHS detection or related experiments contain GFs and are potential nonphysiological sources of the detected GFs in MCs. In addition, heparin also exhibits high binding affinity to avidin and streptavidin, key components of the most widely used IHS detection and amplification system. Although biotin-avidin-free detection systems are readily available and are highly recommended for the future studies in this field, the vast majority of the studies of GFs in MCs in the literature have used biotin avidin-based methods. In this chapter, the inherent technical pitfalls related to the aforementioned features of MCs, and suggested solutions are presented. They are intended to provide technical assistance to investigators in this field and to help interpret the results of the past studies in the literature. PMID- 16273707 TI - Toward a methodology of Reform Jewish bioethics. PMID- 16273708 TI - Portrait of an electronic optometric office. PMID- 16273709 TI - The morality of ovarian transplants. PMID- 16273710 TI - When there is a reluctance to refer patients, everyone loses. PMID- 16273711 TI - Begin year-end tax planning with a look at vehicle purchases. PMID- 16273712 TI - Attitudes of the public and scientists to biotechnology in Japan at the start of 2000. AB - This survey on biotechnology and bioethics was carried out on national random samples of the public and scientists in November 2000-January 2000 [sic]throughout Japan, and attendees at the Novartis Life Science Forum held on 29 September 1999 in Tokyo. The sample size was 297, 370, and 74 respectively. While there is a better awareness of GMOs in 2000 compared to 1991; the trend shows an increase in the perceived risks of GMOs followed by growing resistance in Japan. While a majority of persons believed genetic engineering would make life better over the next twenty years (57%), the proportion of respondents who thought genetic engineering would make life worse over the next twenty years doubled from 1997 to 2000 (from 12% to 25%). Respondents were asked whether they had heard about applications in several areas and the order of familiarity (high low) was: pest-resistant crops, human genes in bacteria, mouse to develop cancer, food and drinks, pigs with human hearts and pre-implantation diagnosis. A divide of opinion can be seen when the results on benefit, risk and moral acceptability of applications of biotechnology by the public are compared to the forum and scientist samples. A significant change in the acceptance of the public occurred in 2000 where only 22% agreed on the moral acceptability of GM food compared to 41% in 1997. In 2000 fewer people said they are willing (20%) to buy genetically modified fruits that taste better compared to 1997 (36%). The results show less public support for use of gene therapy than 1993 and twice as many scientists rejected gene therapy than they did in 1991. When asked who is best placed to regulate modern biotechnology, the respondents were overwhelmingly in favor of international regulatory bodies, such as the United Nations and the World Health Organization (72%), rather than national bodies. The comparison between scientists and public is interesting, however the more enthusiastic sample were participants from the Novaritis Life Science Forum with its mixed occupations. PMID- 16273713 TI - All electronic Medicare fee-for-service claims must now be HIPAA compliant. PMID- 16273714 TI - Anagogy of autonomy. AB - The philosophical and ethical concept of autonomy is herein examined, ex post facto, using an existential lens to examine the process of a personal friend's dying. Anagogy, defined as interpretation of a word, passage, or text that finds beyond the literal, allegorical, and moral senses a fourth and ultimate spiritual or mystical sense, is intended to enlarge the understanding of the use of autonomy in this case. The idea of personhood linked inextricably to reason is, therefore, understood as empowering an individual to choose among various actions, to define and redefine life goals, and to give priority to selected values and moral tenants, which reveal a moral hermeneutic. Conditions and circumstances, existentially exposed, limit choice in unexpected ways, such that the predicted value of autonomy is vulnerable to misuse or misunderstanding. The intent to respect the dignity of every person is central to the philosophy of Respect for Persons ethics, and assumes that autonomy, as freedom of the moral agent, is a moral duty. Implicit reality of freedom is, in a practical sense, essential to being rational agents who can thereby exercise informed choice. The moral law, law of freedom, involves the autonomy of the will and an ultimate end to which all action is directed. Defined as the highest good, morality unites virtue and happiness by ascribing the ultimate end sought as God. The freedom to use rational will finds principles within its own rational nature. The ability to create maxims is autonomy of the will, which equates with the dignity of persons. My recent experience as a companion to a personal friend with a terminal illness inspired me to re-evaluate the concept of autonomy as it is too often interpreted in modern ethical discourse as a individualistic right of choice as opposed to the hermeneutic of dignity of person. This paper describes a shift of position in understanding the paradox of autonomy in this existential context. PMID- 16273715 TI - Immune to marketing? Med students say gifts won't influence them: study. PMID- 16273716 TI - Attitudes and practices of patients and physicians towards patient autonomy: a survey conducted prior to the enactment of the Patients' Rights Bill in Israel. AB - On the surface, it would appear that patients would welcome the opportunity to relinquish their traditional subordination to doctors in therapeutic decision making, and that doctors would be pleased to have partners with whom to share the burden involved in making such fateful decisions. We investigated the attitudes and practices of patients and physicians towards "patient autonomy" in an outpatient clinic of an internal medicine department prior to the enactment of the Patient's Rights Bill in Israel. There were 81 patients randomly chosen from those attending the study clinic and 21 physicians randomly selected from among the physicians treating them. They were all administered the Krantz, the Abramson Health Index, and the Christie Ethical Decision Making pre-tested questionnaires. They were also queried on demographic and background material. The results indicated that the patient sample was neither particularly interested in participating in medical decision making (average score of 3 out of 9 in the Krantz behavioral involvement sub-scale) nor in receiving medical information (average score of 4 out of 7 in the Krantz preference of information sub-scale). The physicians exhibited a willingness to establish equal relations with their patients, and claimed to prefer their taking an active role in decision making. However, when presented with ethical dilemmas, the physicians were not consistent in their attitude in terms of respecting "patient autonomy." The findings of an Israeli survey conducted three years after the bill's passage showed that only one-third of the studied physicians had read the Israel Medical Association booklet's explaining the new law and most of them claimed that the new law had no affect on their daily encounter with patients, meaning that the law did not affect any change in these physicians' pattern of behavior. We concluded that if the Patient's Rights Bill is to achieve its goals, it will have to be accompanied by a widespread educational campaign to encourage the public to appreciate the value and the importance of the autonomy granted to them, and to guide them in exercising this autonomy to its best advantage. In parallel, the medical profession will need to be aware of the importance of achieving the therapeutic goals while upholding ethical and moral values in health care. PMID- 16273718 TI - [Rectal metastases of breast carcinoma]. PMID- 16273717 TI - RNA reigns in neurons. AB - A workshop entitled "RNA Control of Neuronal Function" was recently held in Kfar Blum, Israel. The main topics discussed at the meeting included neuronal RNA targeting mechanisms and the contributing codes and components, translational control mechanisms in dendrites and axons, and the relevance of these mechanisms for neuronal development, plasticity, and dysfunction. PMID- 16273719 TI - [In the memory of E. L. Berezov (the 100(th) anniversary of birth)]. PMID- 16273720 TI - Management of neuropathic pain. AB - Pain may be the most common reason patients seek treatment from physicians. When persistent and unrelieved, pain can frustrate both the person suffering with this condition and the physician trying to alleviate it. Relief from chronic pain may be particularly difficult to achieve and fraught with misconceptions. Treatment usually requires trials of physical, pharmacologic, and surgical interventions to achieve resolution. In cases that remain insoluble, patients must accept partial relief and seek adaptive strategies. The source of persistent pain may be nociceptive or neuropathic. Both utilize the same nervous system pathways for transmission, but significant physiologic differences exist in the mechanism through which the body processes and resolves these painful stimuli. Nociceptive pain that results from a known or obvious source (eg, trauma, cancer metastasis, ischemia, arthritis) is often easy to identify. Neuropathic pain, however, may occur in the absence of an identifiable precipitating cause. Physicians must remain alert to differences in presentation and course of neuropathic pain syndromes, some of which may be subtle or unusual. PMID- 16273721 TI - Update: West Nile virus activity--United States, 2005. AB - This report summarizes West Nile virus (WNV) surveillance data reported to CDC through ArboNET as of 3 a.m. Mountain Standard Time, November 1, 2005. PMID- 16273722 TI - Treatment of Epstein-Barr virus--associated lymphoproliferative disorder (EBV PTLD) and pure red cell aplasia (PRCA) with Rituximab following unrelated cord blood transplantation: a case report and literature review. AB - OBJECTIVE: We report the first case of EBV-PTLD and pure red cell aplasia (PRCA) after an unrelated cord blood transplant in China, who was successfully treated with Rituximab. METHODS: Case report and literature review. RESULTS: A 5-year-old girl with CML underwent a major ABO-incompatible HLA-identical unrelated cord blood transplantation (U-CBT) in January 2003. The post transplant course was complicated by PRCA. She presented on day +75 with fever, followed by rapid enlargement of tonsils, and a cluster of lymph nodes in the cervical and submandibular regions. A cervical lymph node biopsy revealed the histopathologic findings consistent with PTLD. The immunoblasts were shown to contain EBV viral genomic DNA by PCR, and immunocytochemistry study for the latent membrane protein 1 (LMP-1) and in situ hybridization for Epstein-Barr encoded RNAs1 (EBER1) were both positive. She responded rapidly to Rituximab and achieved complete resolution of clinical findings and symptoms of both EBV-PTLD and PRCA. CONCLUSION: EBV-PTLD may occur with increasing frequency due to the increasing numbers of transplant recipients, transplant physician should be aware of this life-threatening complication. Rituximab alone may be an effective therapeutic strategy for patients who develop EBV-PTLD and PRCA after U-CBT. PMID- 16273723 TI - Primary duodenal diffuse large cell non-hodgkin lymphoma with involvement of ampulla of Vater: report of 3 cases. AB - Primary gastrointestinal system lymphomas constitute about one third of all extranodal lymphomas. Duodenal involvement of the lymphoma is a rare condition. Periampullary lymphoma or lymphomatous involvement of ampulla of Vater is even rarer. Since, periampullary lymphoma is not easy to differentiate from epithelial carcinoma of these sites clinically and radiologically, accurate histopathological diagnosis is essential to plan optimal treatment strategy. Obstructive jaundice and the need for some form of drainage procedure and dose modification of the chemotherapy are additional challenges. In this report, we present three cases of duodenal diffuse large cell lymphoma with involvement of ampulla of Vater, two of whom presented with the initial signs of obstructive jaundice. One of the icteric patients was only diagnosed histopathologically following an explorative laparotomy with the initial diagnosis of carcinoma. PMID- 16273724 TI - Intravenous immunoglobulin given to lymphoma patients with recurrent haemolytic transfusion reactions after transfusion of compatible blood. AB - Accelerated destruction of red cells after transfusion of compatible blood has been reported in both sickle cell disease (SCD) and non-SCD patients. We report three patients with lymphoma, all of whom had recurrent haemolytic transfusion reactions after receiving compatible red cell units. The direct antiglobulin test (DAT) was negative and there were no detectable red cell alloantibodies in either pre-transfusion or post-transfusion samples. As there was no evidence of red cell antibody-mediated haemolysis and response to oral steroids, a trial of intravenous immunoglobulin (IVIg) was given. Immediate cessation of haemolysis with sustained haemoglobin level was achieved in all cases. The response to IVIg in these cases suggests that IVIg should be tried when recurrent non-antibody mediated haemolytic transfusion reactions occur in patients with a lymphoid malignancy. PMID- 16273725 TI - Philadelphia chromosome positive acute lymphoblastic leukemia showing normal karyotype in G-banding chromosomal examination before chemotherapy. AB - A 29-year-old male was admitted because of thrombocytopenia. A diagnosis of acute lymphoblastic leukaemia was made on the basis of a 61.6% infiltration of leukemic cells in his bone marrow. Standard G-binding chromosome analysis of bone marrow cells revealed a normal karyotype. He received combination chemotherapy, and achieved hematological complete remission. However, chromosomal analysis of bone marrow cells after 2 courses of consolidation therapy showed the Philadelphia (Ph) chromosome in two cells out of 20 analysed. We retrospectively examined the sample of bone marrow cells before chemotherapy; It showed minor BCR/ABL positivity with FISH and RT-PCR methods. The Ph chromosome disappeared after consolidation chemotherapy and allogeneic bone marrow transplantation, but the Ph chromosome reappeared at relapse. We postulated that there were two clones, both a Ph-positive clone and Ph-negative clone. At the initial diagnosis, Ph chromosome was not detected because the G-banding method analyzed only metaphase cells, which contained few Ph-positive clones. In order to offer effective therapy with molecular targeting agents, in this poor prognostic disease, it is necessary to detect Ph chromosome before the first chemotherapy and BCR/ABL detection with FISH or RT-PCR methods appears more useful than G-banding chromosome analysis. PMID- 16273726 TI - Complete resolution of hepatic aspergillosis after non-myeloablative hematopoietic stem cell transplantation in a patient with acute myeloid leukemia. AB - Fungal infections due to Aspergillus are a frequent cause of transplant-related mortality. For this reason, leukemic patients with severe fungal infection are usually excluded from conventional allotransplantation. Recently, some authors suggested a role for non-myeloablative hematopoietic stem cell transplantation (HSCT) in this subset of patients. We used this therapeutic approach in a patient with high-risk acute myeloid leukemia in second complete remission (CR) with pre existing hepatic aspergillosis refractory to conventional anti-fungal therapy. A complete regression of hepatic lesions was observed after 3 months from allogeneic stem cell transplantation. Our work confirms previous reports suggesting that non-myeloablative HSCT is effective in patients not eligible for conventional transplantation because of invasive aspergillosis. PMID- 16273727 TI - The TCR Vbeta repertoire usage of T-cells from cord blood induced by chronic myelogenous leukemia associated antigen. AB - Understanding the clonality and restricted usage of the TCR Vbeta repertoire of expanded T-cells induced by the chronic myelogenous leukemia (CML) associated antigen may be useful in helping design new immunotherapeutic strategies specifically for CML. T-cells from cord blood that had been stimulated by different stimulators (IL-2, PHA, CML cells, K562 cells and bcr-abl peptide) were amplified in vitro by liquid T-cell culture and the mixed lymphocyte and tumor cell culture (MLTC) method. By using the RT-PCR, the CDR3 segments of 24 variable region genes of TCR beta was analyzed in T-cells from 22 cases of cord blood before and after T-cell culture, to observe the usage of TCR Vbeta repertoire. The PCR products were further labeled with fluorescence and analyzed by the Genescan technique for the CDR3 size, to evaluating clonality of the detectable TCR Vbeta T-cells. Only a part of 24 Vbeta subfamily T-cells (3-15 subfamilies) could be detected, however, all of the 24 TCR Vbeta subfamily of T-cells were detected after in vitro culture with PHA or IL-2+anti-CD3 antibody. The number of expressed TCR Vbeta subfamilies was gradually reduced by prolonging the time of culture with CML-associated antigens. The restricted expression of TCR Vbeta subfamilies and oligoclonal expansion of Vbeta21 T-cells were found in cultured T cells induced by CML cells, K562 cells or bcr-abl peptide. In conclusion, T-cells from cord blood may have the potential capability of proliferation in different TCR Vbeta subfamily T-cells, and the ability for restricted expression and clonal expansion, when T-cells were induced by CML associated antigen. PMID- 16273728 TI - Schiavo under the law. PMID- 16273729 TI - Side effects of corticosteroid therapy in children with chronic idiopathic thrombocytopenic purpura. AB - In 29 symptomatic children of both sexes with chronic idiopathic thrombocytopenic purpura (CITP) with platelet counts < or = 30,000/mm3, of mean age 7.2 +/- 1.3 years, the side effects of long-term glucocorticoid therapy were evaluated. Patients were divided into three groups based on the different glucocorticoid protocols they were receiving. Baseline measurements (t0) of height (H) as standard deviation score (SDS), body mass index (BMI) as kg/m2 and bone mineral density (BMD) at the femoral neck (FBMD) and lumbar spine L2-L4 (LBMD) by a dual energy X-ray absorption technique, expressed as Z score, with follow-up measurements at 1 (t1) and 2 (t2) years were assessed. Group I patients (10 pts) treated with oral prednisone (2 mg/kg/die-for one month for 2 cycles) showed significantly different HSDS, BMI, FBMD and LBMD at t1 and t2 (P < 0.005) than other groups treated respectively with pulsed high doses of dexamethasone (24 mg/m2 over 4 days/months for 6 cycles) and methylprednisolone (9 mg/kg/die for 5 days for 3-4 months). These findings suggest that pulsed high doses of glucocorticoid lead to fewer side effects than oral prednisone therapy. PMID- 16273730 TI - Thrombocytopenia due to cytomegalovirus infection in an immunocompetent adult. AB - A healthy 33-year-old man presented with a short history of viral infection and recent petechiae. Examination of the blood showed a profound thrombocytopenia. He was diagnosed as having cytomegalovirus infection by serological examination. A high level of platelet-associated immunoglobulins was demonstrated suggesting an immune basis for the thrombocytopenia and we, therefore, treated the patient with bolus methylprednisolone and gamma-globulin, resulting in prompt recovery. Recognition for CMV-induced thrombocytopenia is important for early diagnosis. PMID- 16273731 TI - Schiavo under the law. PMID- 16273732 TI - Schiavo under the law. PMID- 16273734 TI - Mormon bioethics. PMID- 16273733 TI - Impact of excess weight and estrogen receptor gene polymorphisms on clinical course of homozygous beta thalassemia. AB - In homozygous beta thalassemic patients we examined the role played by the interaction of ER gene polymorphisms with adverse environmental factors. A total of 108 homozygous beta thalassemic patients, 60 prepubertal mean age 9,5 +/- 3,7 years (27 M, 33 F) and 48 pubertal mean age 22,2 +/- 5,4 years (21 M, 27 F), regularly treated with red cell transfusion and iron chelation therapy were segregated on the basis of their XbaI and PvuII ER gene polymorphisms. Body mass index (BMI), lipidic pattern and blood pressure values were evaluated in each group. No significant differences were observed between patients segregated by their PvuII ER genotypes. Prepubertal and pubertal patients of both sexes lacking XbaI site showed BMI, HDL, LDL cholesterol significantly different than the other patients. In addition, triglyceride levels and blood pressure values were significantly higher in pubertal patients of both sexes lacking XbaI site than in other patients. ER XbaI polymorphism appear to influence nutritional factors, metabolic status and blood pressure and could be considered additional risk factors for later cardiac involvement in beta thalassemic patients. PMID- 16273736 TI - Granulocytic adhesive interactions and their role in sickle cell vaso-occlusion. AB - Recent research has high-lighted the importance of leukocytes in sickle cell disease (SCD). Here we summarize evidence to show that the granulocytes may play a role in SCD due to their increased numbers and adhesive properties, facilitating their participation in the vaso-occlusive process. PMID- 16273735 TI - Acquired haemoglobin H disease. AB - Acquired haemoglobin H disease has been described in various premalignant haematological conditions and is most commonly associated with myelodysplastic and myeloproliferative syndromes. The condition is not restricted to any specific population group or geography. Affected individuals have no family or past history of alpha thalassaemia and these subjects usually suffer from severe uncompensated haemolysis. Extensive mapping and sequence analysis of the alpha globin gene cluster have demonstrated intact alpha globin genes, leading workers to conclude that an acquired in trans mechanism is responsible for the disorder. ATRX gene mutations on the X chromosome have been shown to be instrumental in the suppression of alpha globin gene expression. Despite recent advances in the understanding of its pathogenesis, the precise mechanism of acquired haemoglobin H disease remains a mystery. PMID- 16273737 TI - Investigation of persistent hypochromic microcytosis unmasks hemoglobin Evanston [alpha 14 (A12) Try--> Arg] in a patient of cyclic thrombocytopenia preceding Takayasu's disease. AB - We report an unusual north Indian patient with Hemoglobin Evanston [alpha 14 (A12) Try --> Arg] who had acquired cyclic thrombocytopenia (10-1230 x 10(9)/l periodic oscillation of four week duration) which recovered without any specific therapy. She later developed Takayasu's disease and underwent three corrective stents. She is presently in clinical remission and is on regular follow up. To the best of our knowledge our patient is the first report of Hb Evanston from the indigenous population of India and highlights the need to look for point mutations in the alpha globin gene, which may interact with thalassemia or other hemoglobinopathies, in atypical cases. The association of these three disorders in our patient is possibly unrelated though an immune basis for the cyclic thrombocytopenia and Takayasu's disease is likely as seen in this report. PMID- 16273738 TI - Discussing the theological grounds of moral principles. AB - Discussing the theological beliefs that ground Catholic moral principles can make some people uncomfortable, even while others will appreciate it. But these reactions will sometimes be revealed not as the emotions they are, but as objections to the relative independence or dependence of morality on foundational beliefs. In the end, context should dictate whether one displays the theological beliefs that ground Catholic moral principles. PMID- 16273739 TI - [Polycythemia vera]. PMID- 16273740 TI - Anti HCV seroprevalence among the voluntary blood donors in Thailand. AB - Hepatitis C virus (HCV) infection is an increasing problem, affecting large numbers of the population in both the developed and the developing parts of the world. It is generally accepted as a significant public health problem with major associated morbidities and mortalities, in particular hepatocellular carcinoma. One of several strategies for prevention of HCV transmission is screening for anti HCV serology among donated blood in the blood bank. However, screening for HCV infection varies considerably throughout the world; differences between resource-poor and resource-rich countries are particularly pronounced. This is a particular problem in the developing countries in Asia where HCV infection has high prevalence, this is especially the case among the underprivileged populations and those countries are usually less able to afford routine HCV serological screening in blood bank. In Thailand, HCV infection is an important infectious disease. However, screening for anti HCV serology is performed in only a few large blood banks. The purpose of this study is to summarize the prevalence of Anti HCV seropositivity among the voluntary blood donors in the previous reports in Thailand. This review identified 5 reports in the literature reporting data in 39,633 documented voluntary donors. In all there were 541 cases with Anti HCV seropositivity. The summative percentage for Anti HCV seropositive was 1.37%. This rate is similar to the high levels reported previously from many other Asian countries, implying the importance of HCV infection in Asia. Thorough donor screening to eliminate high-risk donors is recommended to improve blood transfusion services in Thailand and other developing countries and screening for Anti HCV serology should be set as the national strategies covering all blood banks. PMID- 16273741 TI - Discontinuing implantable cardiac devices & the ERDs. AB - Ethics committees are use [sic] to questions concerning the withdrawal of life support. Such questions become increasingly complex when that life-support is implantable, like a pacemaker. This essay seeks to address the question of under what, if any, circumstances it would be permissible to discontinue the use of such implantable devices. PMID- 16273742 TI - A 45-year-old male with fever and hepatosplenomegaly. PMID- 16273743 TI - Nontraditional sources of pluripotent stem cells: a new chapter in the debate about embryonic stem cell research. AB - Recent efforts to resolve the political impasse over human embryonic stem cells (ESC) have generated proposals for obtaining ESC while avoiding the destruction of human embryos. This new chapter in the scientific and ethical debate provides an important opportunity to introduce additional ethical considerations to enhance public discourse. PMID- 16273744 TI - Honoring experience in moral discourse. AB - This essay seeks to address the role of human experience in moral discourse. The authors propose that it is precisely through incorporating a collective experience of what it means to be human in moral discourse that we come to understand the relevance of moral norms in ethical decision-making. We conclude by offering a number of recommendations for ethics committees and hospital administrators. PMID- 16273745 TI - Requests for inappropriate treatment: can a doctor "just say no"? AB - This essay examines (1) the underlying philosophical considerations when patients or decision makers request "inappropriate treatment"; (2) questions to consider in determining if the treatment sought would be ineffective, or, in the words of Weijer et al., effective toward a controversial end; and (3) practical ways to resolve such conflicts. PMID- 16273746 TI - Patients' rights. Final rule. AB - This final rule amends Department of Veterans Affairs (VA) medical regulations to update the patients' rights regulation by bringing its provisions regarding medication, restraints, and seclusion into conformity with current law and practice. The changes are primarily intended to clarify that it is permissible for VA patients to receive medication prescribed by any appropriate health care professional authorized to prescribe medication, and that it is permissible for any authorized licensed health care professional to order the use of restraints and seclusion when necessary. The rule also makes nonsubstantive changes in the patients' rights regulation for purposes of clarification. PMID- 16273747 TI - Medical devices; general and plastic surgery devices; classification of the low energy ultrasound wound cleaner. Final rule. AB - The Food and Drug Administration (FDA) is classifying the low energy ultrasound wound cleaner into class II (special controls). The special control that will apply to the device is the guidance document entitled "Class II Special Controls Guidance Document: Low Energy Ultrasound Wound Cleaner." The agency is taking this action in response to a petition submitted under the Federal Food, Drug, and Cosmetic Act (the act) as amended by the Medical Device Amendments of 1976, the Safe Medical Devices Act of 1990, and the Food and Drug Administration Modernization Act of 1997 (FDAMA). The agency is classifying this device into class II (special controls) in order to provide a reasonable assurance of safety and effectiveness of the device. Elsewhere in this issue of the Federal Register, FDA is announcing the availability of the guidance document that will serve as the special control for the class II device. PMID- 16273748 TI - Medicare program; e-prescribing and the prescription drug program. Final rule. AB - This final rule adopts standards for an electronic prescription drug program under Title I of the Medicare Prescription Drug, Improvement, and Modernization Act of 2003 (MMA). These standards will be the foundation standards or the first set of final uniform standards for an electronic prescription drug program under the MMA, and represent the first step in our incremental approach to adopting final foundation standards that are consistent with the MMA objectives of patient safety, quality of care, and efficiencies and cost savings in the delivery of care. PMID- 16273749 TI - [Antiplatelet drugs and gastrointestinal bleeding: numquam periclum sine periclo vincitur]. PMID- 16273750 TI - [Right ventricular pacing: a resource or a threat?]. AB - Early after the beginning of the pacemaker era, endocardial right ventricular apex has been the most extensively used site for cardiac pacing because it was easily accessible and reliable in a long-term perspective. However many data have demonstrated that this kind of pacing is suboptimal from a physiologic point of view because it causes several adverse effects such as altered ventricular contraction geometry, mitral regurgitation, perfusion alterations and interference with myocardial ion channels which determine a worsening of left ventricular function. Several strategies have been proposed to solve these problems (alternative pacing sites, specific algorithms able to reduce the percentage of ventricular pacing) which are still under evaluation. In this review we analyzed the effects of right apical ventricular pacing and its possible alternatives. PMID- 16273751 TI - [Deleterious effects of apical right ventricular stimulation. Should we change our standard method of pacemaker implantation?]. AB - Up to now the apical right ventricle one is the best pacemaker implantation. As a matter of fact this site is easily reachable by catheter and dislocations are few. Nevertheless evidence from the literature demonstrates dyssynchrony in myocardial contraction pattern, diastolic dysfunction and mismatch in perfusion and innervation. For this reason alternative sites of stimulation have been tested. One of these is represented by the right ventricular outflow tract. Some studies have compared this site to the apical one, showing a better cardiac index in the former; moreover QRS was narrower and fewer perfusional defects have been found. On the contrary, other studies did not show any significant differences between these two sites of stimulation. In order to obtain cardiac resynchronization, biventricular pacing, has been introduced, consisting in the contemporary stimulation of the lateral wall of both ventricles from a cardiac vein, originating from the coronary sinus. It has been proposed a bifocal stimulation, in which we introduce one catheter into the apex and another one in the right ventricular outflow tract: in this case QRS complex is narrower but cardiac output is not increased. A newer pacing technique is represented by direct His bundle stimulation. We can obtain a narrow QRS complex, like the physiological one. So we might solve problems related to intraventricular dyssynchrony. PMID- 16273752 TI - [Isolated left ventricular non-compaction]. AB - Isolated left ventricular noncompaction is a genetically heterogeneous congenital disorder characterized by an altered structure of the myocardial wall. This cardiomyopathy is thought to be due to an arrest of intrauterine compaction of the myocardial fibers in the absence of any other structural heart disease. Noncompaction of the left ventricular myocardium is an uncommon finding and remains frequently overlooked even by experienced echocardiographers. However, a correct diagnosis of noncompaction has important implications due to the possible association with other cardiac abnormalities and/or muscle disorders, progressive left ventricular dysfunction, risk of thromboembolism, and life-threatening arrhythmias. Furthermore, because of the familial association described with ventricular noncompaction, screening with echocardiography of first relatives is recommended. Since echocardiography is the diagnostic technique of choice, missed diagnoses may be due to nonoptimal imaging of the lateral and apical myocardium, and/or insufficient disease awareness by echocardiographers. To increase awareness of left ventricular noncompaction, the present paper reviews embryology, genetics, clinical features and pathophysiology, diagnosis, treatment and prognosis of patients affected by isolated left ventricular noncompaction. PMID- 16273753 TI - [Coronary flow reserve assessment by transthoracic color Doppler echocardiography after primary angioplasty: relationship with recovery of left ventricular function]. AB - BACKGROUND: The aim of this study was to determine the relationship between coronary flow reserve measurement by transthoracic Doppler echocardiography in recent acute myocardial infarction treated with primary coronary angioplasty (PTCA) and recovery of left ventricular function. METHODS: Forty-one consecutive patients (3 patients excluded for not good quality of the Doppler signal) have been studied with: (1) recent first acute myocardial infarction treated with primary PTCA within 6 hours of pain onset; (2) optimal angioplasty result with stent deployment, anti-IIb/IIIa infusion and TIMI 3 flow; (3) lack of type 1 diabetes and/or hypertension; (4) good tolerance to adenosine. Transthoracic Doppler echocardiography was used to record coronary flow velocities in the distal left anterior descending and posterior descending coronary arteries at rest and after infusion of adenosine. Coronary flow reserve was measured after 11 +/- 1 days from the acute event. The wall motion score index (WMSI) was calculated at baseline, 1 month and 3 months from myocardial infarction. RESULTS: Patients of group A (n = 29 with coronary flow reserve > or = 1.6) showed a progressive and significant recovery of left ventricular function at follow-up. Patients of group B (n = 9 with coronary flow reserve < 1.6) had persistent left ventricular dysfunction at 3 months (ANOVA, p < 0.0001). WMSI was 1.64 +/- 0.26 in group A and 1.81 +/- 0.16 in group B (p = 0.09) at baseline; 1.30 +/- 0.26 in group A and 1.75 +/- 0.16 in group B (p < 0.0001) at 1 month; and 1.20 +/- 0.25 in group A and 1.73 +/- 0.17 in group B at 3 months. There was an inverse correlation between coronary flow reserve and WMSI at 1 month (r = -0.564, p < 0.0001), and at 3 months (r = -0.583, p < 0.0001). On multivariate analysis baseline WMSI and coronary flow reserve were the only predictors of 1-month WMSI recovery and of WMSI recovery at 3 months. CONCLUSIONS: Coronary flow reserve by transthoracic color Doppler echocardiography is a useful method for predicting left ventricular function recovery in patients after primary PTCA. PMID- 16273754 TI - [Italian national register of major coronary events: attack rates and fatality in different areas of the country]. AB - BACKGROUND: The national register is a monitoring surveillance system of fatal and non-fatal events in the general population aged 35-74 years; it was launched in Italy at the end of the 1990's with the aim of estimating the occurrence and fatality of coronary events in different geographical areas of the country. METHODS: Two sources of information were used to assess current events: death certificates and hospital discharge diagnosis registers. Once the events were identified through the International Classification for Diseases-ICD9 codes and the duration of the event, the number of current events in each single area was multiplied for the positive predictive value of each specific mortality or discharge code derived from suspected events validated by applying the MONICA Project diagnostic criteria. The attack rate was calculated as the mean value of a 2-year period, dividing the average number of estimated events by the average resident population; case fatality was calculated at 28 days from admission as fatal to total event ratio. RESULTS: Attack rates are higher in men than in women: mean age-adjusted (Italian population 1998) attack rate of all areas was 33.9 per 10,000 men and 9.1 per 10,000 women; age-adjusted 28-day case fatality was higher in women (35.5%) than in men (27.3%). Statistically significant geographical differences in comparison with the mean attack rate of all areas were found both in men and women. Case fatality rates result significantly heterogeneous when compared among areas in men but not in women. CONCLUSIONS: Results show that there still exist some differences in the geographic distribution of attack rate and fatality of coronary events which seem to be independent of the North-South gradient. These data show the feasibility of implementing a population-based register, essential for cardiovascular disease surveillance. PMID- 16273756 TI - [Observational research: a fundamental tool for clinical practice]. PMID- 16273755 TI - [Cardiac surgery in octogenarians: a six-year follow-up with a multidimensional intervention]. AB - BACKGROUND: Elderly subjects frequently experience a decline in function following hospitalization and surgery. Specific changes in the provision of acute hospital care can improve the ability of acutely ill older patients to perform activities of daily living at the time of discharge and the quality of life. The aim of this study was to investigate outcomes of older (age > or =80 years) cardiac surgery patients managed with multicomponent intervention. METHODS: Between 1998 and 2004, we studied records of 193 octogenarian patients who underwent cardiac surgery and were treated with a multicomponent intervention that included: specially designed environment, patient-centered care, planning for patient discharge at home, and an interdisciplinary approach that incorporates in- and out-of-hospital health professionals. RESULTS: Mean follow up was 26.4 months and 100% complete. Mean age of patients was 82.3 +/- 2 years. Eighty-nine patients had myocardial revascularization (CABG), 40 aortic valve replacement (AVR), 34 AVR + CABG, 8 mitral valve replacement (MVR), 11 MVR + CABG and 11 other interventions. Rates of hospital death, major complications and prolonged stay (> 14 days) were as follows: CABG 4 (4.4%), 3 (3.3%), 6 (6.4%); AVR 1 (2.5%), 3 (7.5%), 2 (5%); AVR + CABG 1 (2.9%), 2 (5.8%), 4 (11.7%); MVR 0 (0%), 0 (0%), 1 (12.5%); MVR + CABG 2 (18.1%), 2 (18.1%), 3 (27.2%). Multivariate predictors of hospital deaths were NYHA class, cardiopulmonary bypass and cross clamping time, urgent procedure and ischemic mitral valve procedures. The actuarial 6-year survival was as follows: CABG 91%,AVR 92.5%, AVR + CABG 88.2%, MVR + CABG 81.8%. Total survival rate, free from rehospitalization and redo surgery, was 89.7, 69.8 and 99% respectively. Multivariate predictors of late death were urgent procedure and ischemic mitral valve procedures. At follow-up NYHA classification had improved a median of two classes. Global patients' satisfaction was excellent in 76.7% of survivors; 95.7% were autonomous, 40.5% live at home, 64% had a light-moderate physical activity, and 70% of patients had good social relationships and quality of life. Medical therapy was reduced in 29.3% and level of anxiety improved in 76%. CONCLUSIONS: An interdisciplinary approach and multicomponent intervention with an appropriate postoperative care, provides beneficial effects on outcome in geriatric cardiac surgery patients. PMID- 16273757 TI - Behcet's disease and other autoinflammatory conditions: what's in a name? PMID- 16273758 TI - The rise and fall of FMF research--fifty years of publications. PMID- 16273759 TI - Fibromyalgia in Behcet's disease is associated with anxiety and depression, and not with disease activity. AB - OBJECTIVE: To determine the prevalence of fibromyalgia (FM) in Korean patients with Behcet's disease (BD) and to evaluate the association between FM and clinical and psychological variables. METHODS: Seventy patients with BD were examined for FM tender points and asked to complete a Korean version of the Fibromyalgia Impact Questionnaire (FIQ). Disease activity was measured using the erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), and a clinical activity score, which was calculated by summing the clinical manifestations. The State-Trait Anxiety Inventory (STAI) and Beck Depression Inventory (BDI) were used for psychometric scoring. RESULTS: Twenty-six BD patients (37.1%) met the American College of Rheumatology criteria for FM. The patients who met the criteria for FM were more frequently female, less frequently employed, and less well educated. Age, disease duration, clinical manifestations, medication, and measures of disease activity did not differ between BD patients with and without FM. Nevertheless, BD patients with FM had higher STAI and BDI scores than did patients without FM (all p < 0.05). FM tender points were significantly correlated with the STAI and BDI, and not with disease activity variables. The FIQ scores were also strongly correlated with the STAI and BDI scores, and not with disease activity. CONCLUSION: FM was very common among BD patients and was associated with the presence of anxiety and depression, and not with disease activity. PMID- 16273760 TI - Cytokines, cytokine antagonists and soluble adhesion molecules in patients with ocular Behcet's disease treated with human recombinant interferon-alpha2a. Results of an open study and review of the literature. AB - OBJECTIVE: To elucidate the influence that interferon-alpha exerts on the cytokine network in active ocular Behcet's disease (BD). METHODS: Fifty patients with active ocular BD were treated with human recombinant interferon-alpha2a (rhIFN-alpha2a). Serum was analysed for the presence of IL-10, TNF-alpha, IL-8, IL-6, sIL-2R, IFN-gamma, IFN-alpha, IL-12, IL-4, sTNFRI (p55), sTNFRII (p75), IL 1RA, G-CSF, sE-selectin, sVCAM-1, sICAM-1 and neopterin before initiation of and at several time points during IFN treatment and compared to 21 healthy controls. RESULTS: The levels of IFN-alpha IL1-RA and sTNFRII were significantly increased in the patients at baseline in comparison to healthy controls. During treatment with rhIFN-alpha2a, when remission was achieved as defined by the scoring system used, a significant increase in levels of IFN-alpha, IL-2R, TNF-alpha, sTNF-RII, sICAM-1, sVCAM-1, neopterin in the serum was observed, with a tendency towards increased IL-1RA as well. In contrast, leuko- and thrombocyte counts and sE selectin serum levels significantly decreased. Positive correlations were found between IFN dosage or serum levels and sVCAM-I, neopterin, sTNF-RII and sIL-2R, between sVCAM-1, sIL-2R, TNF-alpha, sTNF-RII and neopterin, sICAM-I and sVCAM-1, sIL2-R and sTNF-RII, and, finally, between sIL2-R and sICAM-I. CONCLUSIONS: IFN alpha exerts diverse influences mainly on cytokine antagonists and soluble adhesion molecules. Because sTNF-RII and IL-1RA were increased by IFN-alpha treatment, these might be interesting alternative treatment options in refractory BD. Some of the side-effects of IFN-alpha may be caused by activation of monocytes, which is reflected by an increase in neopterin serum levels. PMID- 16273762 TI - Behcet's disease associated with malignancies. Report of two cases and review of the literature. AB - OBJECTIVE: To investigate the incidence of malignancies in a cohort of Behcet's disease patients and review the world literature. METHODS: Our database of 128 patients was searched and the age standardized rate (ASR) for cancer was calculated. Furthermore, we performed a MEDLINE search from 1970 through 2003, as well as, a search in the proceedings of international conferences for cases of malignancies associated with Behcet's disease. RESULTS: Two of our 128 patients with Behcet's disease were found to have solid tumors. One male had lung cancer and the other female had kidney cancer. The ASR for cancer cases in our population was investigated and it was found to be 1,600 per 100,000 in 10 years. The ASR for cancer cases in Greece according to WHO is 272.51 per 100,000 per year and therefore 2,725 per 100,000 in 10 years. In the world literature 112 cases of malignancies associated with Behcet's disease were found: Sixty five cases were of male patients and 46 of female with 1 case of unknown gender. The solid malignancies associated with Behcet's disease included cases of bladder, breast, uterus, thyroid and stomach cancer, whereas haematological malignancies included leukemia, myelodysplastic syndrome, lymphoma, multiple myeloma, Hodgkin's disease and lymphosarcoma. The treatment administered in these patients with their disease is also reported. CONCLUSION: The age standardized rate of cancer in our population was lower than that of the general population in Greece, although the difference was not statistically significant. However, there is discrepancy in the world literature and the possibility of development of malignancies in Behcet's disease patients should not be ignored. PMID- 16273761 TI - Behcet's disease patients present high levels of deglycosylated anti-lipoteichoic acid IgG and high IL-8 production after lipoteichoic acid stimulation. AB - OBJECTIVES: Lipoteichoic acid (LTA), induces some of the clinical symptoms of Behcet's disease (BD) in a rat animal model. These results led to the hypothesis that LTA may also trigger BD in humans. We investigated the humoral and cellular immune response against LTA and lipopolysaccharide (LPS) in patients with BD, and compared these responses with those of patients with active chronic oral ulcers (OU) and normal controls. METHODS: Samples were obtained from 12 active BD, 12 inactive BD, 12 active OU and 12 normal controls. Anti-LTA, anti-LPS antibodies levels and the capacity of immune complexes anti-LTA IgG-LTA to activate complement were studied. Exposed mannose residues in anti-LTA IgG were analyzed in the four groups. The interleukin-8 (IL-8) production by peripheral blood mononuclear cells cultures after LTA and LPS stimulation was also studied in all groups. RESULTS: The capacity to bind mannan binding protein (MBP) of anti-LTA IgGs was significantly higher in BD and active OU patients relative to normal controls (p < 0.001). However, only active BD patients generated significantly higher levels of C5a than controls (p < 0.0001). The IgGs purified from the sera of BD patients showed a high specificity for LTA from Streptococcus sanguis or Streptococcus faecalis. LTA also stimulates the secretion of IL-8 in peripheral blood mononuclear cells isolated from active BD patients. Anti-LPS IgA and IgG titers were significantly higher only in active OU patients relative to normal controls (p < 0.0018). CONCLUSION: These results suggest a mechanism involving LTA from streptococci in the pathogenesis of BD. PMID- 16273763 TI - Vascular endothelial growth factor and monocyte chemoattractant protein-1 in Behcet's patients with venous thrombosis. AB - OBJECTIVE: Vascular lesions can involve both arterial and venous systems which are often the major causes complicating the disease course of Behcet's disease (BD). Vascular endothelial growth factor (VEGF) is a stimulant of angiogenesis secondary to ischemia while monocyte chemoattractant protein 1 (MCP-1) is induced by shear stresses leading to vascular collateral development. MCP-1 has been also shown to contribute to the recanalization of venous thrombi. Tumor necrosis factor-alpha (TNF-alpha) is known to play a major role in the pathogenesis of BD. Furthermore, up-regulation of secreted MCP-1 and VEGF was observed following stimulation with TNF-alpha. In view of the above functions of VEGF, MCP-1 and TNF alpha, we hypothesized that these factors may be important in the pathogenesis of thrombosis seen in BD. METHODS: A total of 36 patients with a diagnosis of BD were studied. BD patients were separated into 3 groups with respect to vascular involvement. Group BD-AT (n = 9) with acute thrombosis, BD-CT (n = 12) with chronic thrombosis and BD-MC (n = 15) with mucocutaneous involvement only. The control group (group H) was comprised of 20 healthy persons. In addition, patients with acute, DC-AT (n= 11) and patients with chronic DC-CT (n = 9) thrombosis without BD served as disease controls. Serum measurements of VEGF MCP 1 and TNF-alpha were performed by quantitative sandwich ELISA. The acute phase reactants, including erythrocyte sedimentation rate (ESR) and C-reactive protein (CRP) were also measured. RESULTS: The levels of VEGF were significantly higher in the patients in group BD-AT than either in group BD-CT or BD-MC (p = 0.03 and p < 0.001, respectively). However, no significant difference was found for VEGF levels of thrombotic patients regarding the cause (BD-AT vs. DC-AT, p = 0.063; BD CT vs. DC-CT, p = 0.084) or the stage of thrombosis (DC-AT vs. DC-CT, p > 0.05). Both BD patients and disease controls with acute thrombosis had significantly higher levels of MCP-1 as compared to corresponding chronic thrombosis patients (BD-AT vs. DC-CT; p < 0.001; DC-AT vs. DC-CT, p < 0.001). Patients with BD and disease controls had significantly higher serum TNF-alpha level when compared with healthy subjects. No significant difference with respect to serum TNF-alpha level was noted when patient subgroups with BD and disease controls were compared with each other Serum levels of VEGF, MCP-1, and TNF-alpha were not found to be correlated with either ESR or CRP (p > 0.05). CONCLUSIONS: Increased levels of VEGF and MCP-1 detected in BD thrombosis suggest the possible role of those angiogenic cytokines in the pathogenesis. Although not specific for BD, detection of VEGF or MCP-1 levels seems to serve as an assay for differentiation of BD patients with acute thrombosis from chronic. PMID- 16273764 TI - Circulating soluble CD28 in patients with Behcet's disease: relationship to clinical manifestations. AB - OBJECTIVE: This study evaluates the presence of serum soluble CD28 (sCD28) in Behcet's disease (BD) and its relationship with clinical manifestations. METHODS: Soluble CD28 concentration was determined by ELISA in 120 patients with BD (80 patients in active stage), 60 patients with rheumatoid arthritis (RA) and 60 healthy subjects. RESULTS: Concentrations of sCD28 were significantly higher in patients with BD and RA than in healthy subjects. Patients with active BD expressed the highest level of sCD28 in serum. Soluble CD28 exhibited a drastic increase in active BD patients, compared to BD in remission. Soluble CD28 concentrations were higher in patients with active BD patients having vasculitis. Significant positive correlation was observed in a longitudinal study of 15 BD patients, between sCD28 and C-reactive protein. CONCLUSION: Our study suggests that fluctuations of sCD28 in BD reflects disease activity and should be assessed in evaluating disease activity. PMID- 16273765 TI - Fibrinolytic activity and d-dimer levels in Behcet's syndrome. AB - OBJECTIVE: Thrombophlebitis occurs in a third of patients with Behcet's syndrome (BS). The thrombotic tendency in BS has been studied with inconclusive results perhaps due to the inadequate numbers of patients studied during the acute phase of the thrombosis as well as the lack of appropriate diseased controls. We have studied tissue-type plasminogen activator (t-PA) and its inhibitor (PAI-1), and d dimer levels in BS patients with and without thrombosis both in the acute and chronic phases along with suitable diseased and healthy controls. METHODS: t-PA and PAI-1 were studied by ELISA and d-dimer by semiquantitative latex agglutination slide test in 30 BS patients without deep vein thrombosis (DVT), 10 BS with acute DVT (ADVT), 25 BS with chronic DVT, 27 with ankylosing spondylitis, 26 diffuse systemic sclerosis, 15 patients with ADVT due to other causes, 10 patients with sepsis, and 23 healthy controls. RESULTS: The t-PA levels in BS with ADVT were significantly lower than those in patients with ADVT due to other causes (7.4 +/- 6.2 vs. 13.4 +/- 6.3, P = 0.027) while PAI-1 levels did not show significant differences between the groups (P = 0.60). The numbers of patients with d-dimer levels of > or = 0.5 microg/ml in BS with ADVT were similar to those found in patients with ADVT due to other causes (9/10 vs. 14/14). CONCLUSION: The relatively low t-PA levels point to a defect in fibrinolysis in BS. d-dimer levels are increased in the acute phase of thrombosis in BS. PMID- 16273766 TI - Interleukin-18 gene polymorphisms in Korean patients with Behcet's disease. AB - OBJECTIVE: There is strong evidence that Th1-type cytokines play an important role in the pathogenesis of Behcet's disease (BD). Interleukin (IL)-18 is a proinflammatory cytokine that mediates Th1-polarized immune responses, and elevated levels of IL-18 have been observed in the sera and bronchoalveolar lavage fluid of patients with active BD. Therefore, the aim of this study was to investigate the potential associations of two single nucleotide polymorphisms (SNPs) at positions -137 (G/C) and -607 (C/A) in the promoter region of the IL-18 gene with a susceptibility to BD in the Korean population. METHODS: Ninety-eight patients with BD and 105 healthy controls were studied. All of the subjects were genotyped using sequence specific PCR. The genotypes and alleles between patients with BD and controls were compared using the chi2 test, together with Yate's correction where appropriate. Haplotype analysis was assessed using the EH program. RESULTS: The genotype and allele distributions of the two SNPs did not differ significantly between patients with BD and controls. The haplotype frequencies of the IL-18 promoter polymorphisms were also similar between patients with BD and controls. However, the frequency of the GG genotype at position -137 was significantly higher in BD patients with ocular lesions than in those without ocular lesions (p = 0.026, pc = 0.048, OR = 4.1). CONCLUSION: Although the IL-18 gene polymorphisms were not associated with a susceptibility to BD in the Korean population, the patients carrying the GG genotype at position -137 had a higher risk of developing the ocular lesions. Further studies in other populations are required to confirm these results. PMID- 16273767 TI - Behcet's disease and hereditary periodic fever syndromes: casual association or causal relationship? AB - OBJECTIVE: Mutations in the MEFV and the type 1 TNF receptor (TNFRSF 1A) genes have recently been linked to familial Mediterranean fever (FMF) and TNF receptor associated periodic syndrome (TRAPS), respectively. A higher prevalence of Behcet's disease (BD) among FMF patients has been described compared to the general population. The aim of this study was to evaluate whether FMF TRAPS and BD could be genetically related. METHODS: We screened a cohort of 50 BD patients and 100 healthy subjects for the common MEFV and TNFRSF 1A mutations. An initial screening of exons 10 and 2 of the MEFV gene and exon 4 of the TNFRSF 1A was performed in all chromosomes. RESULTS: The heterozygous MEFV mutation (K695R) was found in one (2%) BD patient. Analysis for FMF mutations in the control group revealed that 5 (5%) individuals bore MEFV gene mutations (3 were heterozygous for the E148Q and 2 were heterozygous for the A744S). At codon 202, there were no differences in allele frequencies between BD and control population: 73%R 27%Q in the BD patients vs 75%R 25%Q in controls. Concerning mutations in the TNFRSF 1A gene, the R92Q mutation was present in heterozygous state in one (2%) BD patient and in 4 (4%) controls without differences between allele frequencies: 99%R 1%Q in BD patients vs 98%R 2%Q in controls, respectively. There was no association between the clinical manifestations of BD patients and the presence of a particular polymorphism or a mutation. CONCLUSIONS: Neither FMF nor TRAPS are genetically associated with BD in our cohort of Spanish patients. PMID- 16273768 TI - Anti-Saccharomyces cerevisiae antibodies (ASCA) in Behcet's syndrome. AB - OBJECTIVES: Anti-Saccharomyces cerevisiae antibodies (ASCA) are found in 50-60% of patients with Crohn's disease. Increased as well as normal levels have been reported in Behcet's syndrome (BS). We reassessed the level of IgG and IgA ASCA antibodies in BS and in a group of diseased and healthy controls. METHODS: Eighty five patients with BS were studied along with 20 patients with ankylosing spondylitis (AS), 24 with Crohn's disease (CD), 25 with ulcerative colitis (UC) and 21 healthy volunteers. A commercial ELISA kit was used (Inova Diagnostics). RESULTS: It was only the patients with CD who had significantly higher levels of antibodies compared with the rest of the group (ANOVA: ASCA IgG, p = 0.0001; ASCA IgA, p = 0.0001). 42% of CD, 4% of BS, 4% of UC and 15% of AS patients had a positive IgG+IgA ASCA. There was a significant trend for patients with gastrointestinal (GI) involvement with BS (n = 8) to be more positive for IgG and IgG+IgA ASCA compared to the rest of the patients with BS (n = 77) (Chi-square, IgG, p = 0.02, IgG+IgA, p = 0.001). CONCLUSION: The rate of positivity of ASCA in BS is comparable to that observed among patients with UC and AS. Patients with BS who have GI involvement may have higher levels of ASCA and this needs to be further studied. PMID- 16273769 TI - Increased thickness of the carotid artery intima-media assessed by ultrasonography in Behcet's disease. AB - OBJECTIVE: Behcets disease (BD), is a unique systemic vasculitis, which affects almost all types and sizes of blood vessels. Carotid intima-media thickness (IMT) is an endothelial cell dysfunction (ECD) parameter which may also be associated with atherosclerosis. We aimed to search carotid IMT and plaque formation in BD, using high-resolution B-mode Doppler ultrasonography (USG). METHODS: We studied 114 BD patients (M/F: 68/46; mean age 38.15 +/- 9.44 years; disease duration 121 +/- 79 months), being followed up by Ege University Rheumatology Department. Age and sex-matched, 77 healthy controls, and as the disease control group 46 non matched SLE patients were also included. Exclusion criteria for all the study participants were hypertension, hyperlipidemia, diabetes mellitus, obesity and history of cardiovascular or cerebrovascular disease. Comparison of the three groups were made by ANOVA and for post-hoc confirmation, Bonferoni test was used. RESULTS: The carotid IMT in BD (mean +/- SD, 0.55 +/- 0.14 mm) was significantly higher than in healthy controls (0.48 +/- 0.09 mm) (p = 0.004), but significantly lower than in SLE (0.66 +/- 0.24 mm) (p = 0.001). Likewise, plaque frequency in BD (5/114) was significantly higher than in healthy controls (0/77), but significantly lower than in SLE (8/46) (p < 0.001). CONCLUSION: Despite significantly higher carotid IMT and plaque frequency in BD compared with healthy controls, these parameters in BD were not as marked as in SLE. Less severe carotid artery abnormalities in BD, may partially explain why cardiovascular morbidity and mortality do not seem to be increased in BD, unlike in SLE. PMID- 16273770 TI - Serum interleukin 17 and interleukin 18 levels in familial Mediterranean fever. AB - OBJECTIVE: Familial Mediterranean fever (FMF) attacks are characterized by serosal inflammation rich in PMNL leukocytes and activation of a definite cytokine network. Moreover, there is sustained inflammation in attack-free FMF patients. Interleukin (IL)-17 and IL-18 are recently described proinflammatory cytokines, which can modulate certain neutrophil functions. In this study we measured serum levels of IL-17 and IL-18 in FMF patients. METHODS: The study groups comprised of 18 FMF patients in attack-free period (mean age: 30.2 +/- 9.5 years; male/female: 10/8), and 18 patients with an acute FMF attack (mean age: 25.4 +/- 4.9 years; male/female: 10/8). Twenty age-matched healthy subjects were included as a control group (male/female: 10/10). Levels of IL-17 and IL-18 were determined by commercial ELISA kits (Biosource International, USA). RESULTS: Serum IL-17 levels were 42.8 +/- 3.7, 42.7 +/- 3.2, and 39.9 +/- 2.3 pg/mL for FMF patients in attack-free period, FMF patients with acute attack, and healthy controls, respectively. Serum IL-18 levels were 878.8 +/- 315.0, 854.2 +/- 261.4, and 314.6 +/- 80.8 pg/mL for FMF patients in an attack-free period, FMF patients with acute attack, and healthy controls, respectively. Levels of both IL-17 and IL-18 were significantly higher in FMF patients with and without acute attack compared to control group (p < 0.05). Concentrations of those cytokines were comparable in FMF patients with acute attack and in attack-free period (p > 0.05). CONCLUSION: Our data suggest that IL-17 and IL-18 contribute to the cytokine network in the inflammatory cascade of FMF. However, their roles for the initiation of FMF attacks remain to be established. PMID- 16273771 TI - Oxidative burst response to monosodium urate crystals in patients with Behcet's syndrome. AB - OBJECTIVE: An erythematous response to intradermal injection of monosodium urate crystals (MSU) has been demonstrated in Behcet's syndrome (BS). To further elucidate the pathogenesis of this response, the effects of MSU on in vitro oxidative burst reaction of neutrophils and monocytes were investigated. METHODS: Peripheral blood mononuclear cells from patients with Behcet's syndrome (BS), rheumatoid arthritis (RA), familial Mediterranean fever (FMF) and healthy controls (HC) were incubated with 100 ng/ml phorbol myristate acetate (PMA) and MSU at different dosages (25-500 microg/ml). Oxidative burst reaction was evaluated in neutrophils and monocytes by flow cytometry. RESULTS: In patients with BS, oxidative burst of neutrophils was significantly increased compared to HC at 125 microg/ml and 250 microg/ml dosages of MSU (p < or = 0.001 and 0.004 respectively). In patients with FMF; there was also an increased oxidative burst reaction at 75 microg/ml, 250 g/ml and 500 microg/ml (p < or = 0.007; 0.001 and 0.004 respectively). In patients with BS, oxidative burst of monocytes was increased only at 125 g/ml dosage of MSU (p < or = 0.002). However, in patients with FMF monocyte burst response was increased at 25 microg/ml, 75 microg/ml and 125 g/ml (p < or = 0.004; < 0.0001; < 0.0001 and 0.002 respectively). In RA group, stimulation with PMA resulted in a higher oxidative burst reaction than FMF and BS (p < or = 0.000 and p < or = 0.008). No correlation was observed between oxidative burst of neutrophils or monocytes and intradermal responses to MSU crystals. CONCLUSION: Oxidative burst reaction with MSU is augmented in neutrophils and monocytes of BS. However, the response is not specific and is unassociated with skin dermal test which has a high specificity for BS. PMID- 16273772 TI - The proportional Venn diagram of Behcet's disease-related manifestations among young adult men in Turkey. AB - OBJECTIVES: To determine the frequency of the features associated with Behcet's disease (BD) in a young men population and generate a proportional Venn diagram of those features. METHODS: Data was collected from 3714 otherwise healthy men recruited for military service at the entrance. Study was conducted in a two-step procedure. Firstly, all participants were questioned by a general practitioner via using visual Behcet's questionnaire. Those participants, in whom at least one BD-related manifestation of the disease (oral ulcer, genital ulcer, folliculitis, erythema nodosum, uveitis, venous involvement of the lower extremities) have been demonstrated, were further examined by a rheumatologist at the second-stage of the study. RESULTS: The areas of intersection among the 6 individual BD-related manifestations produced 63 mutually exclusive symptom groups. Sixteen out 63 of these groups were functionally operative in our study population. Forty-seven (1.2%) of the all participants were considered to have at least 1 of the BD related manifestation after examined by rheumatologist. The prevalence rates of the individual manifestations among the study population were as follows; oral ulcer 29 (0.78%), folliculitis 31 (0.83%), genital ulcer 9 (0.24%), venous involvement 13 (0.35%), erythema nodosum 4 (0.10%) and uveitis 3 (0.08%). The group consisting of oral ulcer with folliculitis was the largest proportion of participants followed by the group having oral ulcer only, accounting for 0.29% and 0.18%, respectively. Four (0.1%) of the participants were fulfilled the International Study Group for BD criteria following rheumatologic and ophthalmologic examinations. After excluding the group having oral ulcer with folliculitis, additional 12 cases had features suggesting BD though they didn't fulfill the International Study Group for BD criteria. CONCLUSION: The Venn diagram of this study demonstrates that International Study Group for BD criteria can detect almost the quarter of 16 cases suspected as having BD. We suggest that the application of information regarding the frequencies of individual BD-related manifestations and their association with each other in a general population might serve as a helpful tool for physicians while making diagnosis. PMID- 16273774 TI - Behcet's disease: familial clustering and immunogenetics. AB - Behcet's disease (BD) is a relapsing, multisystemic inflammatory disorder, characterized by major symptoms consisting of recurrent orogenital ulcerations, eye and skin lesions. Other clinical features may include musculoskeletal, vascular, gastrointestinal, renal, cardiopulmonary or neurological involvement. Vasculitis affecting all types and sizes of blood vessels is the main histopathologic process, in a third of cases complicated by thrombosis. The etiopathogenesis is presently unknown, but BD likely represents the result of a peculiar immune response to hitherto unidentified environmental factors in genetically predisposed subjects. The prevalent distribution in a specific geographical area spanning the Mediterranean basin and Asia, the close association with human leukocyte antigen B*51 in different ethnic groups, and the familial clustering of BD are hallmarks accounting for the strong contribution of a genetic background. The BD familial aggregation is characterized by both genetic anticipation and higher prevalence in childhood patients, likely defining a subset with stronger immunogenetic influences. Polymorphisms in genes encoding for host effector molecules may have a supplementary role in disease susceptibility and/or severity. The contribution of prothrombotic mutations and polymorphisms in the pathogenesis of BD thrombosis is controversial. In this paper, the available reports on BD familial clustering and the evidence for the role of immunogenetic predisposing factors are reviewed. PMID- 16273773 TI - Behcet's disease, myelodysplastic syndrome, trisomy 8, gastroenterological involvement--an association. AB - Only a limited number of cases of Behcet's disease and hematological malignancies have been reported in the literature. We report the case of a 45 year old female patient with Behcet's disease who developed myelodysplastic syndrome, refractory anemia with excess blasts in transformation subtype, with complex chromosomal abnormalities, including excess of chromosome 8, following several years of treatment with chlorambucil for Behcet's disease. As has been described in most such cases, gastrointestinal involvement was most prominent. This case is described and the occurrence of myelodysplastic syndrome in Behcet's disease reviewed. PMID- 16273775 TI - Superior vena cava thrombosis and chylothorax in a young patient. PMID- 16273776 TI - Immunosuppressive treatment of AA amyloidosis of familial Mediterranean fever. PMID- 16273777 TI - Behcet's disease associated with trisomy 8 in a male Caucasian patient from Great Britain--a case report. PMID- 16273778 TI - Complexities in the quantitative assessment of patients with rheumatic diseases in clinical trials and clinical care. AB - Quantitative measurement has led to major advances in the diagnosis, prognosis and management of chronic diseases. Quantitative measures in rheumatic diseases differ from measures in many chronic diseases in several respects. There is no single "gold standard," such as blood pressure or cholesterol, in the diagnosis, management, and prognosis of any rheumatic disease. Laboratory tests are limited; for example, in rheumatoid arthritis > 40% of patients or more have a normal erythrocyte sedimentation rate (ESR). Formal joint counts have poor reliability and are not performed at most visits of most patients. Radiographs are rarely read quantitatively, except in formal clinical trials. The optimal quantitative measures to monitor status and assess long-term prognosis are often derived from patient self-report questionnaires. Quantitative measures may reflect disease activity, e.g., swollen joint counts or C-reactive protein (CRP), long-term damage, e.g., radiographic damage, or poor outcomes, e.g., work disability and premature death. Disease activity measures used in clinical trials are primarily surrogates for long-term outcomes. As there is no single "gold standard" measure, indices of multiple measures are used in patient assessment. Indices used in rheumatoid arthritis assess primarily disease activity, but separate indices have been developed to assess disease activity versus damage in patients with ankylosing spondylitis, systemic lupus erythematosus, and vasculitis. PMID- 16273779 TI - OMERACT: an international initiative to improve outcome measurement in rheumatology. AB - OMERACT is the acronym for an international, informally organized network aimed at improving outcome measurement in rheumatology. Chaired by an executive committee it organizes consensus conferences in a 2-yearly cycle that circles the globe since 2002. Data driven recommendations are prepared and updated by expert working groups. Recommendations include core sets of measures for most of the major rheumatologic conditions. Since 2002 patients have been actively engaged in the process. OMERACT 8 will take place in Malta, May 2006 (www.omeract.org). PMID- 16273780 TI - The Health Assessment Questionnaire (HAQ). AB - Patient-reported outcomes (PROs) provide intrinsic knowledge about a patient's health, functional status, symptoms, treatment preferences, satisfaction, and quality of life. They have become an established approach for assessing health outcomes. The Health Assessment Questionnaire (HAQ), introduced in 1980, is among the first PRO instruments designed to represent a model of patient-oriented outcome assessment. The HAQ is based on five patient-centered dimensions: disability, pain, medication effects, costs of care, and mortality. It has been validated by mail, in the office, by telephone, and by comparison with paraprofessional and physician judgments as a reliable instrument, and has been significantly correlated with other PRO instruments. Typically, one of two HAQ versions is used: the Full HAQ, which assesses all five dimensions, and the Short or 2-page HAQ, which contains only the HAQ disability index (HAQ-DI) and the HAQ's patient global and pain visual analog scales (VAS). The HAQ-DI and the global and pain VAS (i.e., the short HAQ) have essentially retained their original content since their inception, while the Full HAQ undergoes periodic revision to address issues of contemporary scientific interest. The HAQ-DI has been translated or culturally adapted into more than 60 different languages or dialects and has become part of the National Institutes of Health "Road-map" Project, the Patient-Reported Outcomes Measurement Information System (PROMIS). PMID- 16273781 TI - Development of a multi-dimensional health assessment questionnaire (MDHAQ) for the infrastructure of standard clinical care. AB - The HAQ has become the pre-eminent patient questionnaire used in rheumatology. It is easily completed by patients, but not easily reviewed and scored in standard clinical care and has some minor psychometric limitations, as do all questionnaires. Modifications of the HAQ been made to facilitate use in standard care, particularly to include 8-10 activities of daily living, along with scores for pain and global status and other information on one side of one page for rapid review by the clinician. A patient questionnaire for standard care should be limited to 2 sides of 1 page, in a format amenable to "eyeball" review by the clinician in 5 seconds or less. It can be scored formally in 15-20 seconds or less, and is useful in patients with all rheumatic diseases. The current version of a multi-dimensional HAQ (MDHAQ) includes scoring templates on the questionnaire to allow formal scoring in less than 15 seconds by a rheumatologist or an assistant, for possible entry onto a paper and/or computerized flow sheet. Various versions of the MDHAQ may also include a "constant" region of physical function, pain and patient global status, and "variable" regions of fatigue, morning stiffness, psychological distress, change in status, a review of systems, a rheumatoid arthritis disease activity self-report joint count (RADAI), review of recent health events, and review of medications. The MDHAQ can be used in the infrastructure of rheumatology care to include quantitative data in standard care of all patients with all rheumatic diseases. PMID- 16273782 TI - Why the HAQ-II can be an effective substitute for the HAQ. AB - The HAQ-II is a psychometrically improved 10-item version of the standard HAQ functional questionnaire. It is simpler and faster to administer and score, thereby reducing patient burden. HAQ-II is better correlated with clinical measures than the HAQ and is sensitive to change. The HAQ-II is suitable for use in the clinic and research studies. PMID- 16273783 TI - Measuring functional disability in early rheumatoid arthritis: the validity, reliability and responsiveness of the Recent-Onset Arthritis Disability (ROAD) index. AB - OBJECTIVE: Disability has been identified as a core outcome measure in rheumatoid arthritis (RA). The aim of this study was to test the Recent-Onset Arthritis Disability (ROAD) questionnaire for validity, reliability and responsiveness in Italian patients with early RA. METHODS: The psychometric properties of ROAD were tested in 159 patients with early RA, mean age 54.7 (+/- 8.8), 74.3% women, mean disease duration 14.5 months (+/- 1.9 months). All completed the ROAD, the Medical Outcomes Study SF-36 Health Survey (SF-36), the Health Assessment Questionnaire (HAQ) and the patient global assessment (PGA) of functional disability twice, in order to test for validity and responsiveness. Of the 159 patients who completed the health status instruments on two occasions, 121 were included in the responsiveness analyses. The test-retest reliability of the ROAD questionnaire was calculated using intraclass correlation coefficients (ICCs) and the Bland and Altman method on 77 patients who completed the questionnaire twice over an interval of one week. Construct validity was assessed using Spearman's correlations, while responsiveness was evaluated by 3 different methods: (1) effect size (the mean difference between the baseline scores and thefollow-up scores divided by the standard deviation of the baseline scores); (2) standardized response mean (the mean change in scores divided by the standard deviation of the change in scores); (3) receiver operating characteristics (ROC) curve analysis. RESULTS: ROAD fulfilled the established criteria for validity, reliability and responsiveness. In comparison with the SF-36, the expected correlations were found when comparing items measuring similar constructs, thus supporting the convergent construct validity. Significant correlations were seen between ROAD scores and HAQ scores (rho = 0.372), SF-36 physical component summary (PCS) (rho = -0.413), PGA functional disability (rho = 0.417), pain (rho = 0.639), Ritchie index (rho = 0.357), number of swollen joints (rho = 0.387), patient and physician assessment of disease activity (rho = 0.467 and 0.323, respectively), and Disease Activity Score (rho = 0.476). Test-retest reliability was satisfactory, with ICCs of 0.927 (upper extremity function), 0.892 (lower extremity function), and 0.851 (activity of daily living/work). Bland-Altman plots confirmed this finding. The results of responsiveness analysis indicate that the ROAD subscales were slightly more sensitive to perceived change in functional disability than those of HAQ, SF-36 PCS, and PGA offunctional disability. CONCLUSION: Our data suggest that the ROAD index is a reliable, valid and responsive tool for measuring physical functioning in patients with early RA, and is suitable for use in clinical trials and daily clinical practice. Its generalizability and utility for assessing aggressive treatment and functional outcomes must now be evaluated in broader settings. PMID- 16273784 TI - The consequences of rheumatoid arthritis: quality of life measures in the individual patient. AB - Despite conventional treatment, RA still has many deleterious consequences. From the patients' perspective, these include persistent pain, functional disability, fatigue, and depression modified by health beliefs and underlying psychological problems. Disability is a consequence of pain, active synovitis and joint damage. It is usually assessed by self-reported questionnaire; the Health Assessment Questionnaire (HAQ) remains the dominant disability measure, although generic health measures such as Short Form-36 and Nottingham Health Profile provide similar information. Treatment with disease modifying drugs and biologic agents improves pain, fatigue and disability. We specifically evaluated the effects of both these drugs and also disease duration on disability assessed by HAQ scores, as there is most information on this topic and it is of fundamental importance to patients. In early RA HAQ gives a 'J-shaped' curve; the initial fall is due to the immediate benefits of treatment and the subsequent gradual rise due to the inability of therapy to fully suppress the disease or prevent progressive joint damage. In established RA HAQ scores increase by about 1% annually and over 25 years average HAQ scores increase by 1.0. Disease modifying drugs and biologics both significantly reduce HAQ scores and the reduction is maintained for 2-5 years. This reduction is seen in both early and established disease. Early steroid therapy has immediate symptomatic treatment, but does not have long-term benefits. Over 5 years the impact of aggressive therapy with disease modifying drugs declines and there is evidence that insufficient treatment is given to many patients with RA. The outcome of RA is greatly improved by current treatment with disease modifying drugs and biologic agents. However, more needs to be done and achieving better results is enhanced by routinely measuring the impact of the disease in routine practice. PMID- 16273786 TI - Quantitative joint assessment in rheumatoid arthritis. AB - A count of swollen and tender joints is the most specific quantitative clinical measure to assess and monitor the status of patients with rheumatoid arthritis. Many methods have been described to quantitate joint abnormalities, including scoring various numbers of joints (with or without grading of abnormality) for different types of abnormalities, including swelling, tenderness, pain on motion, limited motion, and deformity. This article reviews selected methods for the performance of joint counts, with discussion of their advantages and limitations in the assessment of patients with rheumatoid arthritis. PMID- 16273785 TI - The promise of PROMIS: using item response theory to improve assessment of patient-reported outcomes. AB - PROMIS (Patient-Reported-Outcomes Measurement Information System) is an NIH Roadmap network project intended to improve the reliability, validity, and precision of PROs and to provide definitive new instruments that will exceed the capabilities of classic instruments and enable improved outcome measurement for clinical research across all NIH institutes. Item response theory (IRT) measurement models now permit us to transition conventional health status assessment into an era of item banking and computerized adaptive testing (CAT). Item banking uses IRT measurement models and methods to develop item banks from large pools of items from many available questionnaires. IRT allows the reduction and improvement of items and assembles domains of items which are unidimensional and not excessively redundant. CAT provides a model-driven algorithm and software to iteratively select the most informative remaining item in a domain until a desired degree of precision is obtained. Through these approaches the number of patients required for a clinical trial may be reduced while holding statistical power constant. PROMIS tools, expected to improve precision and enable assessment at the individual patient level which should broaden the appeal of PROs, will begin to be available to the general medical community in 2008. PMID- 16273787 TI - Radiographic progression in rheumatoid arthritis. AB - Radiographic progression is an important outcome measure in clinical trials and observational studies with patients with rheuamtoid arthritis. In this article we describe several aspects of measuring radiographic progression. We introduce the scoring method, discuss scoring methodology and issues regarding reliability of scoring, describe the relation between disease activity, radiographic progression and physical function, and introduce the concept of repair, a novelty in the field of measuring structural changes in RA. PMID- 16273788 TI - The use of second generation anti-CCP antibody (anti-CCP2) testing in rheumatoid arthritis--a systematic review. AB - OBJECTIVE: To evaluate the diagnostic properties and predictive value of the second generation of anti-CCP antibodies (anti-CCP2) in rheumatoid arthritis (RA) patients. METHODS: A systematic review of the published literature between January 2002 and June 2005 was performed. Data were extracted regarding the sensitivity and specificity of anti-CCP2 antibodies in making an accurate diagnosis of RA, predicting future development of RA, and predicting future radiological damage in RA patients. In addition, the prevalence of CCP2 antibodies in patients with other rheumatic diseases was examined. RESULTS: Among 38 studies initially identified, 27 provided information on the use of anti-CCP2 testing. Diagnostic properties were assessed in 13 studies; reported sensitivities ranged from 14.4% to 96%, and specificities from 88.9% to 100%. Odds ratios (OR) for the future development of RA varied from 15.9 among previously healthy individuals to 37.8 among a group of patients with undifferentiated arthritis. Several studies suggested that the presence of anti CCP2 antibodies is highly predictive of current radiographic damage and further damage progression. CONCLUSIONS: Anti-CCP2 has a low sensitivity to be used as a screening test. However, a positive test is highly specific for RA. In addition, anti-CCP2 appears to be highly predictive of the future development of RA in both normal individuals and patients with undifferentiated arthritis. Finally, the presence of anti-CCP2 antibodies appears to predict radiographic damage and progression among patients with RA. PMID- 16273789 TI - Assessment of pain in rheumatic diseases. AB - Pain is the most prominent symptom in people with musculoskeletal disorders, and the most common motivation for patients seeking medical help. However, pain generally is not recorded quantitatively in routine medical care. Over the last three decades, self-report questionnaires have been developed in which a patient may record quantitatively a pain score at baseline and over time to determine whether their condition has improved, remains unchanged, or has worsened. The most robust quantitative pain measure appears to be a simple 10 cm visual analog scale (VAS), which can be completed by the patient and scored by a health professional in less than 10 seconds. Quantitative data concerning pain cannot be obtained from any source other than the patient. Quantitative assessment of pain at each visit in routine rheumatology care, along with the assessment of functional disability, global status, and other patient variables, using a patient self-report questionnaire might lead to improved patient care. PMID- 16273790 TI - Rheumatology function tests: quantitative physical measures to monitor morbidity and predict mortality in patients with rheumatic diseases. AB - Physical measures of functional status, including grip strength, walking time and button test, had been used in rheumatology clinical trials for many years, but have been supplanted in recent years by patient questionnaires. While patient questionnaire measures involve minimum professional time and have greater predictive value than physical measures for severe long-term outcomes of rheumatoid arthritis (RA), physical measures bypass socio-cultural differences which may be seen in use of patient questionnaires. Inter-observer and intra observer reliabilities of these physical measures were excellent when administered according to a standard protocol for instructions. Physical measures of function also were significant predictors of mortality in two cohorts of patients with RA, one monitored between 1973 and 1988, and a second between 1985 and 1990. PMID- 16273791 TI - Laboratory monitoring of biologic therapies. AB - The purpose of this report is to provide suggested guidance concerning the monitoring of TNF blocker therapy. Since the completion of randomized trials, several new long-term safety concerns have arisen, involving mycobacterial and opportunistic infections, cytopenias, lymphoma, demyelinating disease, drug induced lupus, congestive heart failure and hepatotoxicity. Since these serious events are rare, widespread post-marketing use and prolonged follow-up have been required to analyze their prevalence. Monitoring of TNF inhibitors is necessary to reassure physicians and patients of the continued efficacy and safety of these drugs. No published recommendations on monitoring are available. The clinician must weigh the potential clinical benefits of TNF inhibition against potential adverse effects. Patients should be evaluated carefully for the risk or presence of infection, tuberculosis and other serious adverse events by regular visits, careful clinical assessments, and an assiduous, high index of suspicion for these rare events. Tuberculin skin testing using PPD is recommended before starting treatment with any TNF inhibitor. PMID- 16273792 TI - The Disease Activity Score and the EULAR response criteria. AB - In rheumatoid arthritis (RA), inflammatory activity cannot be measured using one single variable. For this reason the Disease Activity Score (DAS). has been developed. The DAS is a clinical index of RA disease activity that combines information from swollen joints, tender joints, the acute phase response and general health. The DAS-based European League Against Rheumatism (EULAR) response criteria were developed to measure individual response in clinical trials. The EULAR response criteria classify individual patients as non-, moderate, or good responders, dependent on the extent of change and the level of disease activity reached. PMID- 16273793 TI - The Simplified Disease Activity Index (SDAI) and the Clinical Disease Activity Index (CDAI): a review of their usefulness and validity in rheumatoid arthritis. AB - Composite indices or pooled indices are useful tools for the evaluation of disease activity in patients with rheumatoid arthritis (RA). They allow the integration of various aspects of the disease into a single numerical value, and may therefore facilitate consistent patient care and improve patient compliance, which both can lead to improved outcomes. The Simplified Disease Activity Index (SDAI) and the Clinical Disease Activity Index (CDAI) are two new tools for the evaluation of disease activity in RA. They have been developed to provide physicians and patients with simple and more comprehensible instruments. Moreover, the CDAI is the only composite index that does not incorporate an acute phase response and can therefore be used to conduct a disease activity evaluation essentially anytime and anywhere. These two new tools have not been developed to replace currently available instruments such as the DAS28, but rather to provide options for different environments. The comparative construct, content, and discriminant validity of all three indices--the DAS28, the SDAI, and the CDAI- allow physicians to base their choice of instrument on their infrastructure and their needs, and all of them can also be used in clinical trials. PMID- 16273794 TI - The American College of Rheumatology (ACR) Core Data Set and derivative "patient only" indices to assess rheumatoid arthritis. AB - Pooled indices of several measures have been developed to assess and monitor patients with rheumatoid arthritis in clinical trials and clinical care, as no single measure can serve as a "gold standard" in all individual patients. Early indices of disease activity include the Steinbrocker "therapeutic scorecard in rheumatoid arthritis," the Lansbury Index, and Paulus criteria. The most widely used indices at this time are the American College of Rheumatology (ACR) Core Data Set and disease activity score (DAS). A simplified disease activity index (SDAI) and clinical disease activity index (CDAI) are derived from the DAS. The ACR Core Data Set includes 7 measures--swollen joint count, tender joint count, patient assessment of global status, an acute phase reactant [erythrocyte sedimentation rate (ESR) or C-reactive protein (CRP)], health professional assessment of global status, physical function, and pain; the first four of these measures are included on the DAS. Improvement criteria for the ACR Core Data Set are based on improvement of at least 20% in both tender and swollen joint counts, and three of the five additional measures (ACR 20), and corresponding "ACR 50," and "ACR 70." A pooled index which includes only the three patient self-report questionnaire measures from the Core Data Set, physical function, pain, and patient assessment of global status performs as well as ACR 20 or DAS to discriminate between efficacy of active versus placebo treatment in a clinical trial. PMID- 16273795 TI - A 3-page standard protocol to evaluate rheumatoid arthritis (SPERA): efficient capture of essential data for clinical trials and observational studies. AB - An efficient 3-page format known as the "standard protocol to evaluate rheumatoid arthritis" (SPERA) has been developed to collect essential baseline clinical data in clinical trials and clinical research studies. The three pages address: 1) clinical features of rheumatoid arthritis (RA), 2) medications taken, and 3) a 42 joint count. Two additional documents, a patient questionnaire and a radiographic scoring sheet, are included for a comprehensive database. The 15-20 minutes needed to complete the SPERA generally adds efficiency over time in standard clinical care, and does not preclude the collection of additional information for clinical care and/or clinical research. The SPERA is presented not as the most desirable format, but rather as an example of a possible approach to the development of a consensus in the rheumatology community regarding a common format for the collection of core clinical data in RA. PMID- 16273796 TI - Assessment of systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) is the archetypal autoimmune disease given its complex clinical and molecular manifestations. Like the other rheumatic diseases, appropriate management is critically dependent upon the proper assessment of disease activity, organ damage, and quality of life. Here, we describe the components of the comprehensive assessment of SLE, including accurate physical and laboratory diagnosis, monitoring of disease activity, recording of accumulated organ morbidity, and integration of these with the patient's own perceptions of health status and quality of life. In doing so, we will review the most appropriate laboratory tests and indices currently used in standard clinical care and in clinical research. PMID- 16273797 TI - Assessment of ankylosing spondylitis. AB - Outcome measures for the assessment of patients with ankylosing spondylitis (AS) have been the subject of considerable research in the last decade, largely through the contributions of the ASessments in Ankylosing Spondylitis (ASAS) International Working Group. This review focuses on the measurement of disease activity, physical function and structural damage in AS, both in daily patient care and to measure treatment response in clinical trials. The ASAS Core Sets for assessment in AS are an important tool to guide disease monitoring, and the domains they contain are discussed, along with other possible concepts important to patient care, including imaging and health-related quality of life. In clinical trials, the assessment of disease response to therapy using the ASAS Response Criteria is a valuable means of determining treatment efficacy and allows comparison of response across trials and interventions. PMID- 16273798 TI - The assessment of disease activity and outcomes in psoriatic arthritis. AB - Psoriatic Arthritis (PsA) is a common condition that significantly impacts affected patients. The introduction of novel therapeutic agents for PsA has generated considerable interest in both clinical trials and in clinical care. Thus, there is a great need for standardized outcome measures to assess the activity of disease and the response to therapy. Because psoriasis is a heterogeneous and multi-faceted condition, defining outcome measures has been a challenge. To date, such measures have largely been adapted from related diseases, as described in this essay. Further research is needed to further develop outcome measures for PsA to facilitate optimal treatment of patients with PsA. PMID- 16273799 TI - The WOMAC Knee and Hip Osteoarthritis Indices: development, validation, globalization and influence on the development of the AUSCAN Hand Osteoarthritis Indices. AB - Clinical measurement in both clinical research and clinical practice requires tools and techniques that are valid, reliable and responsive. Patient-centred self-reported measures provide opportunity to evaluate consequences of osteoarthritis, that are important and relevant to patients with the condition. The WOMAC and AUSCAN Indices are health status measurement questionnaires that are valid, reliable and responsive, easy to complete, simple to score and available in multiple language forms and scaling formats. They provide opportunities to capture patient relevant information, relating to the impact of interventions, in clinical research and clinical practice environments. WOMAC data have also contributed to the development of proposed definitions for responder criteria and state-attainment criteria in osteoarthritis. PMID- 16273800 TI - The Fibromyalgia Impact Questionnaire (FIQ): a review of its development, current version, operating characteristics and uses. AB - The Fibromyalgia Impact Questionnaire (FIQ) was developed in the late 1980s by clinicians at Oregon Health & Science University in an attempt to capture the total spectrum of problems related to fibromyalgia and the responses to therapy. It was first published in 1991 and since that time has been extensively used as an index of therapeutic efficacy. Overall, it has been shown to have a credible construct validity, reliable test-retest characteristics and a good sensitivity in demonstrating therapeutic change. The original questionnaire was modified in 1997 and 2002, to reflect ongoing experience with the instrument and to clarify the scoring system. The latest version of the FIQ can be found at the web site of the Oregon Fibromyalgia Foundation (www.myalgia.com/FIQ/FIQ). The FIQ has now been translated into eight languages, and the translated versions have shown operating characteristics similar to the English version. PMID- 16273801 TI - The Arthritis, Rheumatism and Aging Medical Information System (ARAMIS): still young at 30 years. AB - Chronic diseases such as atherosclerosis, arthritis, diabetes, and cancer are among major public health concerns. To understand their cumulative risk factors and antecedents, a chronic disease databank consisting of time-oriented, multidisciplinary longitudinal data, prospectively collected on consecutive patients and describing their clinical courses, provides a systematic anthology of patient reported outcome (PRO) data. ARAMIS, which began in the mid-1970s, was the first large-scale chronic disease data bank system. Outcomes data are collected using the Health Assessment Questionnaire (HAQ), a well established PRO instrument that collects patient-centered data in the areas of disability, pain and other symptoms, adverse effects of treatment, economic impact, and mortality. More than 900 peer-reviewed studies have emanated from ARAMIS since its inception. In the earlier days, and even today, ARAMIS had to invent its own tools for the study of these new sciences. ARAMIS has made dominant contributions to the understanding of PROs and to helping improve treatment and health outcomes in rheumatoid arthritis (RA), osteoarthritis (OA), scleroderma, lupus, aging, and drug side effects. It continues to traverse terrain with participation in the NIH "Roadmap" project, the Patient Reported Outcome Measurement Information System (PROMIS). PROMIS is designed to provide improved assessment of health status across all chronic illnesses as part of an improved infrastructure for clinical research. As initiator of the rich history of chronic disease data banks with "rolling" consecutive open patient cohorts, ARAMIS has enabled the study of real world PROs in rheumatology, with a wealth of resultant improved approaches to treatment, outcome, cost effectiveness, and quality of life. PMID- 16273802 TI - A brief introduction to the National Data Bank for Rheumatic Diseases. AB - The National Data Bank for Rheumatic Diseases (NDB) is a research data bank with a broad agenda and approach to important clinical, regulatory, epidemiological, safety, effectiveness, outcome and patient care questions that cannot be answered by conventional data banks. It has systematic, ongoing quality control programs that assure high quality data. NDB-developed programs result in very efficient analytic capabilities and rapid publication. PMID- 16273803 TI - The CORRONA database. AB - BACKGROUND: Large, long-term databases are needed in order to provide information on the safety and efficacy of new agents used in the treatment of rheumatoid arthritis (RA) and psoriatic arthritis (PsA). These databases can provide data which is well beyond what is available from industry-sponsored investigations. METHODS: The structure, governance, content, context and developmental plan of the CORRONA database is described. RESULTS: The CORRONA database has grown from start up in 2002 to the largest independent database in North America which collects data from both rheumatologists and patients at the time of a clinical encounter. Data are collected as often as every 3 months in RA and every 6 months in PsA. As of the time of this writing, the CORRONA database consists of approximately 9,000 patients with RA and 1,000 with PsA. Data can be used to elucidate toxicities found in frequencies which would be considerably less common than can be uncovered in industry-sponsored investigations. In addition, actual prescribing patterns and responses in clinical practice can be investigated and described. CONCLUSION: After 3 years of data collection, the CORRONA database is now appropriately able to make significant contributions to our understanding of the safety, efficacy of drugs, as well as demographic, and socioeconomic profiles of patients with RA and PsA. It has evolved from a nascent database to a mature one poised to make significant contributions. PMID- 16273804 TI - An Early Rheumatoid Arthritis Treatment Evaluation Registry (ERATER) in the United States. AB - An Early Rheumatoid Arthritis Treatment Evaluation Registry (ERATER) was established in 2001 to enroll patients with a disease duration of 3 years or less, in order to analyze treatment patterns in an era of biological therapies and to study the long-term outcomes of rheumatoid arthritis (RA). Patients were assessed at baseline according to a standard protocol to evaluate their rheumatoid arthritis (SPERA). Similar data from earlier cohorts with RA in the same community will allow for comparisons with treatments and outcomes from previous decades. This essay describes the background regarding the identification of the cohort, methods for data collection, and observations to date. PMID- 16273805 TI - A database in private practice: the Brooklyn Outcomes of Arthritis Rheumatology Database (BOARD). AB - Rheumatologists generally use few quantitative measures in making clinical decisions. In the US, fewer than 10% use questionnaires in routine clinical care, and fewer than 15% perform a formal joint count at each visit. Patient questionnaires are the quantitative tools rheumatologists have to monitor their patients' health status and response to therapy. The health assessment questionnaire (HAQ) and its derivatives have been shown to be the best predictors of functional and work disability, costs, joint replacement surgery and mortality; they are as good as and usually better predictors than joint counts, radiographs and laboratory tests. The Brooklyn Outcomes of Arthritis Registry Database was initiated with the aim of collecting quantitative data using a multi dimensional health assessment questionnire (MDHAQ) from all rheumatology patients seen as part of routine care, each and every time the patient was seen. Data that are feasible to collect in routine clinical care provide the only way to assess quantitatively how our patients are doing. If data are not collected and recorded, an opportunity is lost forever. If there is a reason for the visit, there is a reason to complete a questionnaire. PMID- 16273807 TI - Rheumatoid arthritis registries in Sweden. AB - Patient registries provide valuable contributions to the field of rheumatology for both quality control and scientific purposes. With respect to the latter, patient registries are among the most important datasets used for longitudinal observational studies in rheumatic diseases, which are in turn an essential complement to data obtained from randomized, controlled trials. In Sweden a number of registries are available for such studies, ranging from general medical registries such as the in-patient registry, to rheumatoid arthritis (RA)-specific inception cohorts and biologics registries focusing on a specific patient population defined by a group of treatments. In recent years it has become particularly clear that questions regarding new therapies, their use in practice and their long-term safety, as well as aspects such as pharmacoeconomics, cannot fully be assessed using the data from clinical trials, and that registries are indispensible to obtain accurate answers to such questions. In this review we describe the Swedish rheumatology registries, including the Swedish RA registry and the Swedish biologics registries ARTIS (Antirheumatic Therapies Iin Sweden), SSATG (Southern Sweden Antirheumatic Therapy Group), and STURE (Stockholm Tumor Necrosis Factor-a Follow-up Registry). Data obtained from analyses based on these registries are reviewed. It is concluded that rheumatology registries are excellent tools for improving our knowledge base regarding rheumatic diseases. PMID- 16273806 TI - A Norwegian DMARD register: prescriptions of DMARDs and biological agents to patients with inflammatory rheumatic diseases. AB - Information concerning the effectiveness of drug therapy cannot be obtained only from randomized controlled clinical trials, due to limitations such as a short time frame and narrow inclusion and exclusion criteria. Therefore, complementary longitudinal observational studies performed in a real life setting are required. NOR-DMARD, a Norwegian 5-center register, was established in December 2000. All DMARD prescriptions to patients with inflammatory arthropathies are included, and patients are followed longitudinally with a variety of assessments. As of 2005, 4683 DMARD regimens have been included. Methotrexate is the most commonly used DMARD in rheumatoid arthritis and psoriatic arthritis. The proportions of patients who have received anti-TNF drugs in rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis, juvenile arthritis and other diseases have been 22.5, 21.6, 53.8, 36.9 and 9.7%, respectively. The proportion of patients receiving anti-TNF drugs is considerably higher in 2004 than earlier, and criteria for prescribing anti-TNF drugs appear to be trending toward patients with less severe and active disease. Confounding by indication or channeling bias represents a challenge for the group comparisons of longitudinal effectiveness data, but can be addressed by modern statistical techniques. The NOR-DMARD register may in the future provide comparative real life effectiveness data that may also be used in cost-effectiveness analyses. PMID- 16273808 TI - Rheumatoid arthritis databases in Finland. AB - A tradition exists in Finland of longitudinal studies to analyze the long-term outcomes of rheumatoid arthritis (RA), including early studies in the 1970s and 1980s of work disability and premature mortality in patients with RA and follow up studies of early RA cohorts. This article reviews long-term observations from the Finnish RA Combination Therapy Trial (FIN-RACo), a 2-year multicenter randomized controlled trial which has been continued as a longitudinal observational study after the initial two years. We also describe the Central Finland RA Database, which captures most patients who have been diagnosed with RA since 1980 in a district of a quarter of a million people. PMID- 16273809 TI - DANBIO: a nationwide registry of biological therapies in Denmark. AB - Since the year 2000, Danish rheumatologists have been collecting data on a routine basis in the nationwide DANBIO registry, which includes all rheumatologic patients receiving biological drugs. Demographic data, markers of disease activity, current treatment, serious and non-serious adverse events and reasons for discontinuation are registered at each visit either on paper forms or on line. By June 2005, approximately 3000 treatment courses (18,000 visits) were in the registry, corresponding to close to 90% of eligible patients. Rheumatoid arthritis was the most prevalent diagnosis (75%) followed by ankylosing spondylitis (11%) and psoriatic arthritis (7%). Infections occurred in 43% of the treatment series. PMID- 16273810 TI - Implant occlusion: a literature review. PMID- 16273811 TI - Immediate implant loading of single and multiple implants: where are we now. PMID- 16273813 TI - Mentorship rewards all. PMID- 16273812 TI - Atrophic posterior maxilla and mandible: alveolar ridge reconstruction with mandibular block autografts. PMID- 16273814 TI - Safety of and contraindications for use of intrauterine devices. PMID- 16273815 TI - Planned visits to help patients self-manage chronic conditions. PMID- 16273816 TI - Evaluation of syncope. AB - Though relatively common, syncope is a complex presenting symptom defined by a transient loss of consciousness, usually accompanied by falling, and with spontaneous recovery. Syncope must be carefully differentiated from other conditions that may cause a loss of consciousness or falling. Syncope can be classified into four categories: reflex mediated, cardiac, orthostatic, and cerebrovascular. A cardiac cause of syncope is associated with significantly higher rates of morbidity and mortality than other causes. The evaluation of syncope begins with a careful history, physical examination, and electrocardiography. Additional testing should be based on the initial clinical evaluation. Older patients and those with underlying organic heart disease or abnormal electrocardiograms generally will need additional cardiac evaluation, which may include prolonged electrocardiographic monitoring, echocardiography, and exercise stress testing. When structural heart disease is excluded, tests for neurogenic reflex-mediated syncope, such as head-up tilt-table testing and carotid sinus massage, should be performed. The use of tests such as head computed tomography, magnetic resonance imaging, carotid and transcranial ultrasonography, and electroencephalography to detect cerebrovascular causes of syncope should be reserved for those few patients with syncope whose history suggests a neurologic event or who have focal neurologic signs or symptoms. PMID- 16273817 TI - Supporting self-management in patients with chronic illness. AB - Support of patient self-management is a key component of effective chronic illness care and improved patient outcomes. Self-management support goes beyond traditional knowledge-based patient education to include processes that develop patient problem-solving skills, improve self-efficacy, and support application of knowledge in real-life situations that matter to patients. This approach also encompasses system-focused changes in the primary care environment. Family physicians can support patient self-management by structuring patient-physician interactions to identify problems from the patient perspective, making office environment changes that remove self-management barriers, and providing education individually and through available community self-management resources. The emerging evidence supports the implementation of practice strategies that are conducive to patient self-management and improved patient outcomes among chronically ill patients. PMID- 16273818 TI - Subclinical thyroid disease. AB - Subclinical thyroid dysfunction is defined as an abnormal serum thyroid stimulating hormone level (reference range: 0.45 to 4.50 microU per mL) and free thyroxine and triiodothyronine levels within their reference ranges. The management of subclinical thyroid dysfunction is controversial. The prevalence of subclinical hypothyroidism is about 4 to 8.5 percent, and may be as high as 20 percent in women older than 60 years. Subclinical hyperthyroidism is found in approximately 2 percent of the population. Most national organizations recommend against routine screening of asymptomatic patients, but screening is recommended for high-risk populations. There is good evidence that subclinical hypothyroidism is associated with progression to overt disease. Patients with a serum thyroid stimulating hormone level greater than 10 microU per mL have a higher incidence of elevated serum low-density lipoprotein cholesterol concentrations; however, evidence is lacking for other associations. There is insufficient evidence that treatment of subclinical hypothyroidism is beneficial. A serum thyroid stimulating hormone level of less than 0.1 microU per mL is associated with progression to overt hyperthyroidism, atrial fibrillation, reduced bone mineral density, and cardiac dysfunction. There is little evidence that early treatment alters the clinical course. PMID- 16273819 TI - Genital herpes: a review. AB - Genital herpes simplex virus infection is a recurrent, lifelong disease with no cure. The strongest predictor for infection is a person's number of lifetime sex partners. The natural history includes first-episode mucocutaneous infection, establishment of latency in the dorsal root ganglion, and subsequent reactivation. Most infections are transmitted via asymptomatic viral shedding. Classic outbreaks consist of a skin prodrome and possible constitutional symptoms such as headache, fever, and inguinal lymphadenopathy. As the infection progresses, papules, vesicles on an erythematous base, and erosions appear over hours to days. These lesions usually crust, re-epithelialize, and heal without scarring. First-episode infections are more extensive: primary lesions last two to six weeks versus approximately one week for lesions in recurrent disease. Atypical manifestations are common. Infected persons experience a median of four recurrences per year after their first episode, but rates vary greatly. Genital herpes simplex virus type 2 recurs six times more frequently than type 1. Viral culture is preferred over polymerase chain reaction testing for diagnosis. Serologic testing can be useful in persons with a questionable history. Effective oral antiviral medications are available for initial, episodic, and suppressive therapy but are not a cure. There is some evidence that alternative therapies such as L-lysine, zinc, and some herbal preparations may offer some benefit. Counseling patients about the risk of transmission is crucial and helps prevent the spread of disease and neonatal complications. PMID- 16273820 TI - Information from your family docter. Genital herpes: what you should know. PMID- 16273822 TI - Information from your family doctor. Hidradenitis suppurativa: what you should know. PMID- 16273821 TI - Hidradenitis suppurativa: a treatment challenge. AB - Hidradenitis suppurativa is a chronic, recurrent, debilitating disease that presents with painful, inflamed lesions in the apocrine-gland-bearing areas of the body, most commonly the axillary, inguinal, and anogenital areas. Etiology traditionally has been attributed to occlusion of the apocrine duct by a keratinous plug; however, defects of the follicular epithelium also have been noted. Contributing factors include friction from axillary adiposity, sweat, heat, stress, tight clothing, and genetic and hormonal components. Multiple treatment regimens are available, including antibiotics, retinoids, corticosteroids, incision and drainage, local wound care, local excision, radiation, and laser therapy. However, no single treatment has proved effective for all patients. Radical excision of the defective tissue is the most definitive treatment. The psychological impact on the patient can be great, encompassing social, personal, and occupational challenges. This impact should be addressed in all patients with significant disease. PMID- 16273823 TI - Newborn with a scalp lesion. PMID- 16273824 TI - Information from your family docter. Prenatal diagnosis: amniocentesis and CVS. PMID- 16273825 TI - Information from your family docter. Toxoplasmosis in pregnancy. PMID- 16273827 TI - Time to speak out. PMID- 16273826 TI - Information from your family docter. Respiratory infections during pregnancy. PMID- 16273828 TI - Every child should have an Auntie Mary. PMID- 16273829 TI - Hospital readmissions for preschool viral-wheeze. AB - The prevalence of preschool viral-wheeze (PVW) has increased significantly over the last decade but little is known about the severity of the condition or the incidence of further serious episodes. In order to assess disease severity, number of readmissions, and clinical predictors for readmission we carried out an audit on the clinical notes of 208 children who were admitted with PVW over a six month period. The median duration of hospitalisation was 24 hours. Four children received mechanical ventilation. Forty-six (22 per cent) children were readmitted within six months. No clinical or demographic variable could distinguish the readmitted from the non-readmitted groups. Duration of hospitalisation for PVW is short, but the readmission rate is high: more effective early management and better parental education may be required. PMID- 16273830 TI - Child poverty: is it child abuse? PMID- 16273831 TI - Student assessment using poster presentations. PMID- 16273832 TI - Sexuality in the hospital setting. PMID- 16273833 TI - Staff nurse development programme: evaluation. PMID- 16273834 TI - Ask the children--auditing children's services with PALS standards. AB - Working together is a key element of the NHS Plan (DH 2000). The establishment of the Patient Advice and Liaison Service (PALS) presented staff at the Bournemouth and Christchurch Hospitals NHS Foundation Trust : with an opportunity to examine the experiences of children attending the outpatient department. An audit of outpatient services was undertaken using PALS national standards and the National Service Frame work for Children (DH 2003a). Parents reported receiving good information prior to and during the consultation but would like more written information to take home. The environment was rated positively but older children felt that the waiting area was lacking in appropriate activities to suit their needs. Very few respondents had heard of PALS and around half did not know who to approach with concerns. PMID- 16273835 TI - Inequalities in children's health. AB - This article explores some of the differences between adult and child health services and aims to assist readers appreciate the possible inequalities that can occur. In order to achieve this, different factors affecting child development are explored and the problems of developing policies and practices that support children's health considered. PMID- 16273836 TI - [Job stress in the service industry. Evaluation of the Italian version of the Job Content Questionnaire]. AB - BACKGROUND: Recent changes in work organization have greatly expanded the service industry, where stress perception appears to be the major factor of discomfort. Job stress remains a critical issue in occupational exposure assessments. OBJECTIVES: To assess the compliance and the consistency of the three major constructs (Psychological Job Demand (PJD); Decision Latitude (DL); and Social Support at Work (SSW)) of the Italian version of the Job Content Questionnaire (JCQ) adopted in the MOPSY and JACE studies and their gender-specific distributions in a large sample of civil servant in northern Italy. METHODS: JCQ was administered to 2601 men and 5761 women employed in six different departments of the Municipality of Milan. Two JCQ versions - short and intermediate which differ by the number of items that assess DL for inclusion of SSW, were used in different sub-samples. Compliance was measured in terms of participation rate and percentage of missing items. A construct-specific Cronbach alpha coefficient was used to assess internal consistency. As a measure of external consistency the correlation coefficients between the three constructs were adopted. RESULTS: The participant rate was very high in the entire sample: 75.3% in men and 76.2% in women. The percentage of missing items was small and acceptable, ranging from 3.61% for DL to 6.33% for SSW. The Cronbach alpha coefficients ranged from 0.72 for PJD to 0.88 for SSW. Assumptions for external validity were satisfied in both gender groups. The percentage of perceived high job stress diminished in older men, but the age trend was inverse among women, indicating gender differences in the possibility of improving work conditions and career. Interferences with the family-related load and responsibilities may play a significant role. In both gender groups, a satisfactory SSW level reduced perceived high job stress. CONCLUSIONS: The adopted versions of the JCQ, originally developed in American and north European socio-cultural contexts, demonstrated satisfactory levels of compliance, and internal and external consistency. The JCQ adequately described perceived job stress in the groups of workers under study. PMID- 16273837 TI - [The role of the occupational health physician in smoking cessation programmes: the experience of the "Clinica del Lavoro" of Milan]. AB - <>. BACKGROUND: The "Centre for the prevention, diagnosis and treatment of tobacco dependency", set up at the Occupational and Environmental Respiratory Diseases Section of the Clinica del Lavoro in Milan in September 2000, can count on multidisciplinary expertise, including occupational health physicians, pneumologists and clinical psychologists. Each step of the diagnostic and therapeutic process is based on an integrated approach that considers both the biological aspects of tobacco dependency and the psychological and behavioural aspects. The Centre, moreover, within the framework of "Health Promotion" programmes, offers advice to businesses as regards the new legislation that prohibits smoking at the workplace. METHODS: The proposed treatment programme is based on psychological and motivational support (personal or team work), that can be associated with pharmacological therapy, either as an alternative or an addition. The drug preferred is slow-releasing buproprione, which has proved to be extremely effective compared to other drugs in increasing the probability of smoking cessation and decreasing side effects. RESULTS: In an occupational population of more than 2000 workers, we succeeded in treating about 51% of the subjects, who had stopped smoking, after one year from the beginning of the programme. The best results were achieved using the association of psychological support and buproprione therapy (54%), compared with the other methods (psychological support alone: 42%; psychological support and free nicotine replacement therapy: 33%). PMID- 16273838 TI - [The logo of the Italian Society of Occupational Medicine and Industrial Hygiene]. AB - In 2003 the Board of the Italian Society of Occupational Medicine and Industrial Hygiene (SIMLII) decided to adopt a graphic symbol of the Society, that would be easy recognizable and could be used for the 75th anniversary of its foundation, in 1929. The task of drawing up proposals for the symbol was entrusted to the Section of Occupational Medicine and Toxicology of the University of Perugia. The article explains the difficulties in adequately representing the various different types of information enclosed in the logo; in particular, in choosing the most appropriate way to represent the art of medicine between Mercury's caduceus and Aesculapius' symbol. The logo chosen by the Board of the Society on 11 November, 2003, is illustrated by its author, Professor Umberto Raponi. The symbol officially came into use for the 67th National Congress of SIMLII, held in Sorrento, 3-6 November, 2004. PMID- 16273839 TI - [Radiological and functional progression in silicosis]. AB - BACKGROUND: The incidence of silicosis has decreased today because of a clear improvement of working conditions. According to recent data from Italian National Compensation Agency (INAIL) silicosis represents more than 5% of occupational diseases claimed for annually. Since silicosis is an evolving chronic disease, it has serious consequences on patient health, modifying the quality of life and increasing public costs. Some studies show a relationship between occupational exposure to silica and radiological and functional deterioration, but the results are not satisfactory because of the variety of clinical patterns and the interference of many risk factors. OBJECTIVES: The aim of our investigation was to evaluate silicosis evolution in relation to living habits and to different occupational exposures. METHODS: All admissions from 1980 to 2000 in the Occupational Medicine Unit of the Maugeri Foundation in Cassano Murge (Bari), Southern Italy, with final diagnosis of silicosis were examined. From 586 medical records gathered, subjects with double admissions were selected and their chest radiographs and FVC, FEV1 from lung function test were detected. No industrial hygiene data were available and individual exposure was estimated through calculation of a global exposure index. RESULTS: On the basis of data quality control, 106 consecutive male patients were recruited (age at first admission 51.7 +/- 8.3 years, occupational exposure at second admission 23.8 +/- 9.1 years). More than 50% of the subjects had an estimated high risk exposure at work. The first diagnosis of silicosis was made at first admission in 40% of the cases, 33% of the patients had been suffering from silicosis for 10 years while 27% for more than 10 years. Chest radiographs showed mostly p, q nodular pattern, and over a 4. 7 +/- 3.3 year follow-up period they appeared largely unchanged, although 8 radiographs showed confluent areas, 18 subjects showed pleural thickening, 10 calcified opacities, 17 COPD (Chronic Obstructive Pulmonary Disease) with emphysema, 2 tubercolosis lesions, 4 radiographic abnormalities attributable to cancer (3 lung, 1 oesophageal). CONCLUSIONS: The mainly stationary results of chest radiographs and the slight loss of respiratory function we observed confirm the slow evolution over time of silicosis. Multiple logistic regression analysis of main risk factors showed that subjects with radiographic opacities having a diameter greater than 10 mm or conglomerate shadows at the time of diagnosis seem to have a lower risk of progression of the disease after a relatively short period of follow up, even in mining related exposure. PMID- 16273841 TI - [Characterisation of an HBsAg mutant of hepatitis B virus (HBV) isolated from a dialysed patient involved in an occupational accident]. AB - BACKGROUND: Mutants of the hepatitis B virus (HBV) following vaccination (escape mutants) have been isolated over the course of the last decade. They consist most commonly of an aminoacid change from glycine to arginine at position 145 of the highly antigenic a determinant of the surface antigen (HBsAg). OBJECTIVE: Description of an escape mutant of HBV identified in the course of the post exposure follow-up of a percutaneous exposure. METHODS: The viral DNA was extracted from serum samples of a dialysed patient vaccinated against hepatitis B, who developed an acute infection. A direct sequencing was performed on the amplified DNA followed by a sequence analysis. RESULTS AND CONCLUSIONS: A threonine to lysine substitution at position 118 of HBsAg (Thrll8Lys) was observed in the analysed viral aminoacid sequence. Such mutation could have significantly changed the antigenic profile of the HBsAg compared to that of the wild type. PMID- 16273840 TI - The study of gap junctional intercellular communication in keratinocytes as screening of promoter effect induced by industrial and environmental toxic substances. AB - BACKGROUND: Disordered functioning of gap junctions between normal and initiated cells has been proposed as one possible mechanism of tumour promotion. Many putative carcinogens such as peroxisome proliferators, are known to activate various signal transduction mechanisms and modulate gap junctional intercellular communication (GJIC). They act as tumour promoters on pre-existing "initiated" cells, rather than as genotoxic initiators. OBJECTIVES: The aim of this article is to provide a screening-tool to evaluate the promoter carcinogen effect of environmental and occupational chemical contaminants, focusing on their ability to alter GJIC. METHODS: GJIC was investigated in serum-free cultured primary human keratinocytes, by directly evaluating the intercellular transfer of a microinjected fluorescent dye (Dye transfer). The expression of caspase 3, which is the ultimate target to be activated of both mitochondrial- and non mitochondrial-linked pro-apoptotic pathways, was evaluated using Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR). RESULTS: Mercury chloride (10 nM), mono-methyl Mercury (250 nM) and Trichloroethylene (500 I1M) were shown to significantly inhibit GJIC. Conversely di-methyl mercury, lead acetate and epichloridine had no effect on GJIC. All Trans Retinoic Acid completely reversed the inhibitory effect on GJIC induced by HgCI2 but not that induced by mono methyl mercury and trichloroethylene. The result of a RT-PCR assay on total RNA cell extract showed that treatment of keratinocytes with 10 nM HgCl2 resulted in a decrease of the pro-apoptotic caspase 3 expression. CONCLUSIONS: In this work a protocol is designed to study gap junction intercellular communication in primary cultures of human keratinocytes which could be used as a reliable screening tool to test the promoter carcinogen effect of various environmental and occupational contaminants. PMID- 16273842 TI - [Fatal occupational accidents: updating of data from a mortality register]. AB - BACKGROUND: In Italy, almost one thousand deaths due to occupational accidents are usually registered by INAIL each year. Case registration by INAIL has merely administrative purposes and therefore it is necessary to use other sources for case ascertainment in order to better estimate the real number of deaths related to occupational accidents, as shown also by previous papers. OBJECTIVES: Evaluation of the contribution of another data source, namely the Tuscany Regional Mortality Registry, to obtain the correct figure for occupational accident deaths through the use of a place-of-occurrence notation on the death certificate. METHODS: Cases that occurred in residents in Tuscany in 2000-2001 were considered. They were identified from : a) the Tuscany Regional Mortality Registry (RMR) using the E code of the ICD LX code of death, the year and place of occurrence; b) the INAIL archive using the year of event, the type of definition and management. RESULTS: The INAIL source was without doubt the most informative but was only 51% complete, whereas the RMR source, although less informative, was more complete (82.4%) and allowed identification of cases not registered by INAIL, that had occurred for instance in the Armed Forces and in the National Railway Company. However, the vast majority of RMR extra-cases occurred in subjects aged 65+, in agriculture and in the building industry. CONCLUSION: It is currently possible to plan a systematic linkage of the two sources due to the new possibilities that are available: the place-of-occurrence in the death certificate and the availability of individual data in the INAIL source. PMID- 16273843 TI - [Assessment of asbestos exposure, mortality study, and health intervention in workers formerly exposed to asbestos in a small factory making drying machines for textile finishing and the paper mill industry in Pistoia, Italy]. AB - BACKGROUND AND OBJECTIVES: Three malignant pleural mesotheliomas occurred among workers of a small factory that manufactured drying machines for the textile and paper mill industries using asbestos cement (crocidolite, amosite and chrysotile) as insulating panels. The Occupational Medicine Unit of the Local Health Unit of Pistoia, Italy, carried out an intervention programme in the plant in order to 1) assess past asbestos exposure via analysis of the fibre content of samples from drying machines, and of dust samples collected in the factory. Information on the characteristics of occupational exposure was also collected; 2) investigate cancer mortality by means of a mortality study of the employees and, 3) carry out a health intervention programme in workers formally exposed to asbestos in the past. METHODS: Samples from the drying machines and dust samples collected in the factory were analysed using X-ray diffractometric methods. Information on the characteristics of occupational exposure were collected by interviewing plant workers. Two-hundred and fifty employees who had worked in the factory between 1962 and 2000 were included in the mortality study. Follow-up was performed from 1962 to 2002. Health intervention in workers exposed to asbestos in the past involved general practitioners and occupational physicians (first level medical examinations); pneumologists and radiologists (second level medical examinations) of the local health unit. RESULTS: Asbestos fibres were found both in samples from drying machines and in dust samples collected in the factory. Interviews with workers showed that asbestos exposure varied considerably. The SMR for mesothelioma and lung cancer in 234 male workers were 37.0 (95%CI: 4.47-130.0), and 1.29 (95%CI: 0.26-3.78), respectively, based on mortality rates for Tuscany region. Sixty-two workers underwent first level medical examinations; 57 second level examinations. Chronic obstructive lung disease was found in 3 workers; restrictive lung disease was found in 3 employees, one of whom had pleural plaques. CONCLUSIONS: Further investigation is needed in order to identify unknown asbestos exposures in small metal engineering factories. PMID- 16273844 TI - [Asbestos risk in the textile industry: braking systems on machinery used until the 1990's]. AB - BACKGROUND: We recently described asbestos risk in the non-asbestos textile industry as the result of fibre dispersion from ceilings, pipe insulation and machines. OBJECTIVES: The widespread use of brakes with asbestos linings on the machines as well as other functional details were considered for a proper evaluation of their role in producing atmospheric pollution METHODS: All the information was collected on the basis of the personal technical experience of two of the Authors and by direct observation of the machines. RESULTS: All the textile machines (ring spinning, twisting, warping, winding, looms) used until the 1990's were without exception equipped with asbestos-lined mechanical brakes. The heavy action required produced relatively rapid wear of the linings and the dust produced was spread into the atmosphere by the continuous action of the "travelling blowing cleaners" and by the daily cleaning of the machines using compressed air at the end of the shift: violent air blowing undoubtedly caused redispersion of the fine dust from the brakes and also acted as a mechanical grinder on the bundles that sedimented on the machines from the ceilings and pipes, producing more ultrathin respirable fibres. CONCLUSIONS: the contribution of textile machinery to atmospheric pollution by asbestos fibres was significant and due both to the widespread use of brakes with asbestos-containing materials and to the continuous action on the machines of compressed air blowers. Asbestos pollution was certainly high in all the factories so that in the near future still further mesothelioma cases among ex-workers are to be expected. PMID- 16273845 TI - [Revisiting the normative and medico-legal principles for updating the lists of occupational diseases]. PMID- 16273846 TI - [Asbestos fiber dimensions and mesothelioma]. PMID- 16273847 TI - [Apropos of amianth and mesothelioma]. PMID- 16273848 TI - [Disruption of organizational productivity ("mobbing"): difficulty of intensity and consequences]. PMID- 16273849 TI - [The memory of Professor Antonio Grieco. A founder of contemporary ergonomics in Italy and internationally]. PMID- 16273850 TI - [The national convention "asbestos-related pathology and health surveillance of those formerly exposed" --Pisa, 21-2 April, 2005 ]. PMID- 16273851 TI - Quality of life assessment in otorhinolaryngology. History, measures and methods. AB - The term "quality of life" was first mentioned in 1920 in economic journals and became an issue in the early 1960s in North America. Initially it became accepted in social sciences and later in medicine. The development of generic measures began in the early 1970s and continues today. Disease-specific measures were introduced in the 80th and 90th. During that time methodological studies were implemented and additional emphasis was put on analytic approaches, interpretation of scale scores and cultural including language issues. In ENT study groups began slowly with research in the 1980s and expanded during the 1990s. Today Quality of Life is an accepted part of medical research and covers a broad field of issues. Study groups in ENT work on allergy treatment, oncology and economic aspects of Quality of Life research as well as patient's perception of hearing devices. Future challenges include conceptualization and testing of theoretical models, further refinement of individualized measures for use in routine clinical practice and the inclusion of Quality of Life information in health databases. This article describes the methodological concept, the history of Quality of Life assessment in ENT and gives a detailed overview on current measures. PMID- 16273852 TI - [Some acute phase proteins in children suffering from recurrent laryngeal papillomatosis]. AB - INTRODUCTION: The aim of the study was to compare the character of inflammatory reaction in the groups of sick children, asymptomatic carriers of HPV and healthy children by quantitative and qualitative study of chosen acute phase proteins like CRP, AT, ACT. Profile of glycosylation of ACT was analysed. MATERIAL AND METHODS: The study group was 21 children treated in ENT Department of Pediatric Institute of the Karol Marcinkowski University for laryngeal papillomatosis (infected by HPV 6/11 in 100%) and the group of 22 selected asymptomatic carriers of HPV 6/11. The control group were 22 children in which the presence of HPV 6/11 DNA was excluded using PCR technique. Serum concentration of the APP was measured using immunoelectrophoresis technique according to Laurell. Qualitative study of the APP based on analysis of AGP and ACT microheterogeneity using crossed immuno affinoelectrophoresis on agarose gel according to Bog-Hansen in Mackiewicz modification was carried out. RESULTS: The results were analysed with regard to: 1. the age of the first incident (ten children in whom the first symptoms occurred up to the second year of life, and eleven in whom the first symptoms were detected later; 2. severity of the disease course (seven children in whom four or more microsurgeries were performed within a year, and fourteen children in whom fever than four interventions were carried out). The decrease of ACT, AT levels in serum was observed. It was found that the ACT glycosylation profile is close to normal in sick children, what suggests the absence of inflammatory reaction stimulation. CONCLUSIONS: The results suggest that clinical infection of HPV is likely to be connected with discoordination of immunological reaction mechanism. PMID- 16273853 TI - [Horizontal glottectomy--oncologic and functional results. Part I. Horizontal glottic laryngectomy--oncologic results and assessment of the protective and respiratory function of the larynx]. AB - INTRODUCTION: In the Otolaryngology and Laryngological Oncology Department of the Pomeranian Medical University in Szczecin (Poland) horizontal glottectomy is executed from 1985. MATERIAL AND METHODS: Material consists of 27 patients (26 males and 1 female; an average age 57 min. 40, max.). We evaluated: 1) oncological results, 2) 3 and 5-years survival rate without recurrence of the cancer, 3) evaluation of the protective and respiratory function. Protective function was evaluated basing on: a) period after operation when feeding tube was removed, b) subjective assessment of swallowing liquid and solid food. For evaluating respiratory function we stated period after operation to decannulation and also spirometric investigations were done. RESULTS: From among 27 operated patients died 4 (15%)--one on the second day after operation. Recurrence of the cancer (local or into lymph nodes) were observed in 5/26 (19.2%). Three years without symptoms of recurrence survived 89% (17/19), five years--80% (12/15). Swallowing through natural way was possible during the first 24 hours after operation in 13 of 26 (50%) patients. In the remaining 13 only 2 feeding tubes were removed on the 35th day after operation. Decannulation was possible at 92% (24/26) patients. 15 (58%) patients were decannulated in the first two week after operation. Spirometric investigation (n = 10) showed no restriction and very small obturation on the larynx - very good laryngeal flow nearing to normal. CONCLUSIONS: Oncologic results, protective and respiratory function after horizontal function results are satisfactory and comparable to the other researchers. PMID- 16273854 TI - [Cefprozil in the treatment of chronic maxillary sinusitis. Clinical and microbiological effectiveness and penetration into sinuses examination]. AB - 40 patients with exacerbation of a chronic maxillary sinusitis were examined. Sinus puncture was performed (sinoject) in all of the patients. Before the treatment (500 mg cefprozil orally twice a day, the recommended duration of therapy was ten days) and on the fifth day, the microorganisms from sinus were isolated (the bacteriological culture, antibiogram and MIC were determined). On the second and the fifth day, blood cefprozil level and the presence of cefprozil in the washings from maxillary sinuses were investigated. RESULTS: 17 pathogens were isolated from maxillary sinuses, only two of them were anaerobic. The bacterial strains: Staphylococcus (55%), E.coli (15%), Klebsiella (10%) and 20% of others, were found. Four of them (23.5%) were resistant to cefprozil. In all patients cefprozil was identified in maxillary sinuses (0.87-2.52 ug/ml). The value of MIC were from 0.094 to 2.0 ug/ml. A satisfactory clinical response was observed in 92.5% persons but the eradication of pathogens was obtained in 70%. The adverse clinical events (diarrhea) were observed only in one patient (2.5%). CONCLUSIONS: cefprozil well penetrates into inflammable mucous membrane of maxillary sinuses. Cefprozil obtains efficient bactericidal concentration in relation to sensitive bacteria on the fifth day of therapy. It is also well tolerated by the patients. PMID- 16273855 TI - [Epithelial cell proliferation in nasal polyps]. AB - The mechanism stimulating the growth of nasal polyps remains unclear. The fact that nasal polyps formation is connected with increase of the epithelial surface suggests that the dysregulation of epithelial cell proliferation takes part in their pathogenesis. Furthermore, macroscopically nonsuspected nasal polyp, usually benign tissue lesion, occasionally may be misdiagnosed such as neoplasm. Only a few studies have undertaken the subject of cell proliferation in nonneoplastic respiratory epithelium. We examined 30 nasal polyps specimens by immunohistochemistry to recognize the expression of nuclear antigen present in proliferating cells, using rabbit anti-human Ki-67 antigen. Our study revealed significantly higher Ki-67 index in nasal polyps than in nasal mucosa (p < 0.005, tab.I). Based on this findings it seems that increased epithelial cell proliferation and chronic inflammation generate the lesion of the nasal mucosa leading to nasal polyps formation. PMID- 16273856 TI - [The impact of paraglottic space for laryngeal cancer spread]. AB - INTRODUCTION: Malignant laryngeal cancer is the most common tumor of the head and neck. Radical laryngectomy not only causes the changes in breathing routes and disability to retain the air in the chest during the physical effort, but it also deprives the patient of a possibility to communicate. In order to limit and avoid mutilating surgery i.e. total laryngectomy, precise evaluation of tumor dimensions and spread is indispensable for organ preservation techniques application. AIM: The authors have undertaken the task of estimating the role of paraglottic space in spread of larynx cancer. MATERIALS: The material of 180 patients operated on because of larynx cancer were analysed. 32% patients suffered from epiglottic carcinoma, 26% from glottic, 17% from transglottic carcinoma and 27% from multifocal carcinoma. Subglottic carcinomas was not identified. CONCLUSIONS: The paraglottic space was the most frequently penetrated by advanced tumors invading all three levels of the larynx, next by the transglottic tumors and by glottic localisations of cancer. The difficulties in diagnosing of this anatomical region have been stressed. PMID- 16273857 TI - [Changes of laryngeal mucosa in the course of pemphigus vulgaris]. AB - Blister diseases are chronic autoimmune reactions connected with formation of intraepithelial blisters. Pemphigus vulgaris (PV) change is appear most often, almost 80% of all cases. Erosions on mucosa appear as first symptoms at 50-70% patients. Blisters occurring on the skin are typical for this illness and usually come into with weeks or months with delay in relation to the changes on mucous membranes. In this work we have described character and location of changes on mucous membranes at 5 patients with PV, diagnosed based on clinical symptoms and confirmed in immunofluorescent investigations. PMID- 16273858 TI - [Nasopharyngeal and middle ear flora in children with acute otitis media]. AB - Nasopharyngeal flora can be a reservoir of bacteria caused acute otitis media in children. The aim of the study was to identify microorganisms and antimicrobial susceptibilities of pathogens from the nasopharynx and middle ear of children with acute otitis media. The study comprised 128 children ages 1 year to 14 years with diagnosed of acute otitis media with purulent discharge. The nasopharyngeal and middle ear samples were collected at the same time. Agar, chocolate, blood and Chapman plates were inoculated for isolation of bacteria. The plates were incubated at 37 degrees C and examined at 24 hours. The susceptibility of bacteria was determined by disk diffusion technique containing concentration gradients for following antibiotics: penicillin, amoxicillin/clavulanate, ampicillin/sulbactam, cefaclor, cefprozil, cefuroxime, erythromycin, azithromycin, clindamycin and trimethoprim/sulfamethoxazole. 196 organisms from nasopharynx and 325 organisms from middle ear were isolated. Most frequent cultured bacteria were: Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis--75.6% in nasopharynx and 77.8% in middle ear. We observed statistically significant (p < 0.01) increased of Moraxella catarrhalis in specimens from the middle ear than from nasopharynx. Most of the organisms were susceptible to amoxicillin/clavulanate--83.2% of bacteria from nasopharynx and 81.8% of bacteria from middle ear. Most organisms were resistant to trimethoprim/sulfamethoxazole--60.7% of bacteria from nasopharynx and 62.6% of bacteria from middle ear. Penicillin resistance was observed in 25.0% of bacteria from nasopharynx and 25.6% of bacteria from middle ear. The correlation in resistance of bacteria between trimethoprim/sulfamethoxazole and erythromycin (r = 0.4886) and between trimethoprim/sulfamethoxazole and penicillin (r = 0.5027) was observed. Nasopharyngeal and middle ear flora in children with acute otitis media is similar. In that case susceptibility of bacteria from the nasopharynx can be useful for empirical treatment of acute otitis media in children. PMID- 16273859 TI - [Evaluation of adenoid mast cells in children with secretory otitis media]. AB - Mast cells may play a role in pathogenesis of the otitis media with effusion (OME) in children. The study involved 72 children with no history of allergic diseases. The analysed material were adenoids removed on the grounds of hypertrophy. Immunohistochemical analyses were carried out using antibodies (Dako, M 7052) directed against mast cell tryptase. The slides evaluation was performed by means of optic microscope. The presence of mast cells within adenoidal tissue were scored. There were no statistically significant difference between the numbers of mast cells in each examined adenoid as well as age groups. Our results correspond with those obtained by other authors according to mast cells distribution in adenoid tissue and do not indicate any particular role of mast cells in pathogenesis of OME in children. PMID- 16273860 TI - [Distant metastases to the parotid gland--review of the literature and report of own two cases]. AB - Metastatic tumors to the parotid gland are very uncommon and it accounts for 8% of all cancers of parotid gland. The parotid gland and its lymph nodes are possible sites of metastases from head and neck cancers. However, metastasis from distant primary neoplasm below clavicle is possible, too. The authors presented the two cases of distant metastases to the parotid gland. The women with breast cancer and metastases to the parotid gland 11 years after surgery, radio- and chemotherapy of breast cancer. She died 7 months after parotid surgery of systemically advanced disease. The man with metastasis of malignant melanoma of unknown primary site. He underwent total parotid surgery with the tumor and lymph nodes. He died 7 months after surgery. In spite of intensive exam the primary location of the melanoma was unknown. The authors described pathophysiology of distant metastases to the parotid gland with special attention to possibilities of treatment and survival. PMID- 16273861 TI - [Glucocorticosteroids in the treatment of subglottic laryngitis in children]. AB - Subglottic laryngitis is one of the acute children's diseases, directly caused by a violently growing edema of the subglottic area. Its symptoms generally appear very suddenly, when children seem to be in perfect health, at night, several hours after falling asleep. Their symptoms included barking cough, clear voice, stridor, inspiratory dyspnoea with participation of auxiliary respiratory muscles, excitation and anxiety of a child, changes in skin coloration. The movement of the wings of the nostrils is intensified. In especially severe cases, agitation, cyanosis, pallor of skin, obnubilation, apnea, loss of consciousness and circulatory failure may also occur. Subglottic laryngitis is a disease, which can threaten the life of a small child. The aim of this study was to observe efficacy of the treatment of the subglottic laryngitis with glucocorticoids, especially budesonide in nebulization. The research covered 169 children: 58 girls (34.31%) and 111 boys (65.69%) aged 9. months do 5. years (mean 3 years 6 months) hospitalized in the Children's Hospital in Warsaw with the following symptoms: dry barking cough, stridor, inspiratory dyspnoea with the participation of auxiliary respiratory muscles, agitation and change of colour of skin. The examination of each patient included subjective, objective (pediatric and laryngological). Disease severity was assessed by a clinical croup score based on stridor, cough retractions, dyspnoea and cyanosis and the overall clinical assessment was scored on a visual scale. The results indicate that nebulised budesonide can be used as a safe and effective alternative treatment in children with moderate to severe subglottic laryngitis. PMID- 16273862 TI - [Lymphoproliferative disorders in Sjogren's syndrome]. AB - INTRODUCTION: Sjogren's syndrome [SS] is an autoimmune disease that mainly affects the exocrine glands. B-cell lymphoproliferation is a characteristic feature of this syndrome and the lesion may range from benign to malignant. MATERIAL AND METHODS: After a systematic search of Pubmed we reviewed literature regarding the histopathology, pathophysiology and clinics of lymphoproliferation in SS. RESULTS: Patients with Sjogren's syndrome [SS] have over 40-fold increased risk of the development B-cell non-Hodgkin's lymphoma. Most cases of lymphomas complicating the course of SS arise in mucosal extranodal sites, especially in the salivary gland, and are classified as low grade marginal zone B-cell lymphoma with long-term survival. The main problem in salivary lymphoproliferation in Sjogren's syndrome consists in the difficulties in the differential diagnosis of lymphoma. Genotypic studies have documented the rearrangement of immunoglobulin genes across the full spectrum of lymphoid infiltrates in the salivary gland including cases regarded as reactive lymphoepithelial sialadenitis [LESA], borderline cases with halos of monocytoid cells surrounding epimyoepithelial islets, and cases with fully developed marginal zone lymphoma [MZL]. Thus, the simple detection of B-cell clonality cannot be used as a criterion for the diagnosis of B-cell malignancy. Broad strands of monocytoid B-cells that surround and invade epimyoepithelial islets and monotypic immunoglobulin expression detected by immunohistochemistry are an essential feature for the histopathological diagnosis of MZL. The pathophysiology of lymphoma in SS remains still unknown. Viral infection, hyperstimulation of B cells, disregulation in the process of apoptosis, and unknown oncogenes are suspected to initiate the start of lymphoma. The main clinical features associated with the development of lymphoma in SS include persistent major salivary gland enlargement (> 2 months), persistent lymphadenopathy or splenomegaly, monoclonal gammapathy and type II mixed cryoglobulinemia. The treatment and prognosis of lymphoma associated with SS depend on the type and stage of lymphoma. CONCLUSION: Patients with SS develop a variety B lymphoproliferative disorders. The nature of these must be determined by multiparameter analysis including clinical, histopathological, immunohistochemical and genotypic studies. PMID- 16273863 TI - [Usefulness of ventilation tests in the diagnosis and evaluation of treatment results in select laryngeal diseases]. AB - The obstruction of upper airways is a common symptom of some laryngological diseases. The patients need exact diagnosis and proper treatment. The ventilation tests seem to be one of the most important tool in process of diagnosis and treatment, however they are more pulmonological than laryngological procedures. The aim of the study was to evaluate the efficacy of ventilation tests and analysis of their interpretation in patients with laryngeal and/or tracheal obstruction. Also, we intended to show a method of spirometric examination in patients with tracheostoma. Based on our common clinical experience (more than 3000 tests in patients with upper airways obstruction in ENT Departments in Zabrze and Szczecin) we presented our observations. In majority, the tests were used in patients after laryngeal surgery due to the cancer or bilateral vocal cord paralysis and in patients with tracheostomy, before decannulation. The attention was paid on usefulness of some coefficients, especially in diagnosis of upper airway obstruction. For better results interpretation the authors introduced the term of mixed obstruction in contrast to fixed and dynamically variable narrowing. Also the FIV1/N FEV1 parameter was purposed. In conclusion the great role of ventilation tests was emphasized, especially in diagnosis of the narrowing on the larynx and trachea level and also in evaluation of the results after surgical procedures on upper airways. PMID- 16273865 TI - [Use of plate osteosynthesis as an effective method in surgical treatment of congenital cranial anomalies]. AB - Operative treatment of developmental anomalies were presented in actual work carried out in Maxillo-Facial Clinic PMA in Szczecin. Mesiocclusion was the most frequent anomaly, followed by microgenia, joint ankylosis of tempora mandibular, next laterogenia and first and second branchial arch syndrome. The operative treatment was finished by placing and fixing the bones in new changed position. The obtained results indicated that this type of management is the best effective in correction of visceral cranium development anomalies, it assure quick maintenance of good oral hygiene as well as stabilisation and functioning jaws. PMID- 16273864 TI - [Cystic fibrosis (CF) of adults--current problem of laryngologists. The role of genetic research]. AB - INTRODUCTION: The goal of this work is to evaluate DNA in the direction of gene mutation in the patients with nasal polyps and/or recurring inflammation of the sinuses, and who tested negative for allergic basis of those ailments. MATERIAL AND METHODS: 50 patients aged 20-45 have been included in the research by the University Hospital and the Laryngological Outpatient Clinic SPSK in Bialystok. The patients had symptoms of nasal polyps and recurring inflammation of the sinuses. The analysis of DNA samples of the blood according to the PCR method using the ABI Prism 3100 device and testing for finding mutations of cystic fibrosis--The Cystic Fibrosis Assay. RESULTS: We discovered 1 mutation of delta F508 in a 40-year-old patient, with recurring inflammation of nasal sinuses (described in the case of this patient). CONCLUSIONS: The DNA research method can be used for discovering atypical forms of CF in patients with recurring symptoms of the inflammation of the sinuses and nasal polyps, and who tested negative for allergic basis of those ailments. A wide routine screening in the direction of the mutation of CF may lead to an explanation the reason of the nasal polyps and inflammation of the sinuses. PMID- 16273866 TI - [Rhinophyma--diagnosis and treatment]. AB - The authors present four cases with rare disease--rhinophyma. These disease occurs mainly in males of middle and old age. It results from long-standing rosacea in some patients. Risk factors are blood vessels disorders, endocrinal abnormality, digestive tract disorder. In laryngological practice there are many methods of non-surgical treatment of rhinophyma. The aim of the treatment is to remove deformation and restore function of the nose and prevent the relapse of rhinophyma. Rhinophyma is removed with the epidermis and the wound is realest to self epithelisation in case of small changes, but in more severe cases of rhinophyma the wound needs to be covered by skin scalp. PMID- 16273867 TI - [Smell and taste in drug addicts]. AB - The main objective of this study was to establish the degree of influence of various illicit drugs, taken into the body in different ways, on the senses of smell and taste. Other possible factors, that might have caused disturbances in normal functioning of both senses, were outlined. The studied group consisted of 48 drug addicts, between the age of 16 and 48 years, addicted to various illicit drugs for several years. Olfactory testing included two methods. First quantitative method described by Elsberg and Levy, modified by Pruszewicz, was used to establish the smell detection threshold. Then the University of Pennsylvania Smell Identification Test was used as a method for odor identification. The sense of taste was tested with the use of method described by Krarup and modified by Pruszewicz. The results showed disturbances in olfactory performance (combined in both method) in 52.1% of all drug abusers, and 16.7% of them were diagnosed with ageusia when the sense of taste was tested. The route of administration of illicit drugs proved to be a statistically significant factor that might have caused disturbances of smell perception and identification in the study group. Drug abusers who were taking drugs intravenously and those who smoked and inhaled various drugs had the most significant olfactory problems. Both means for assessing smell function were statistically compared and the results showed that approximately 75% of drug users with olfactory disturbances had olfactory problems in both tests. PMID- 16273868 TI - [Evaluation of phonemic hearing development in CI-patients]. AB - Phonemic hearing it is a typical interdisciplinary problem. Many specialists like pedagogues, psychologists, phoniatricians and speech therapists are interested in this field. So, we may find many definitions describing this kind of hearing. The speech perception and acquisition of speech is a difficult process profoundly hearing impaired children. With the development of the cochlear implant, acoustic input can be provide to these children. Studies documenting the postoperative performance of children with implants have generally shown significant improvements in speech perception and production. On the opposite side--in the group of postlingually deafened adult the speech rehabilitation is quite short. Only a few specialists described their observation about phonemic hearing development in CI-patients. How do it differ from hearing development in healthy population? The aim of this study was to describe the phonemic hearing development in CI-patients. In the ENT Dept. of University of Medical Sciences in Poznan 280 cochlear implantation were performed. They were observe in three groups: pre-, peri- and postlingually deafened ones. We concluded the same order of phonemic hearing development in every group. The time of acquisition differed, and depended of oral language development before implantation. At first they start to distinct voiced and unvoiced phones, than oral-nasal and sharp-soft ones. The most difficult for them was to discriminate fricative and plosive sounds. PMID- 16273869 TI - [Results of hearing perception after cochlear implantation in patients with auditory neuropathy]. AB - INTRODUCTION: Results of hearing perception with patient with auditory and visual neuropathy combined with peripheral polyneuropathy one year after cochlear implantation are presented in this article. Visual problems and hearing loss were progressing during recent years leading to social problems and communication disability. Speech discrimination was much worse than could be suspected based on pure tone audiometry. Lack of benefit from hearing aids was also stated. METHODOLOGY: As the examination method free field speech audiometry was used based on Pruszewicz word tests and set of modified speech therapy tests TAPS II. Simple sentences, monosyllables and polysyllables words as well as logotoms perception and identification of music instruments sounds were estimated. MAIN RESULT: Progress of speech intelligibility in monosyllables test according to Pruszewicz in free field speech audiometry was stated, as well as better understanding of sentences, monosyllables and polysyllables words, and more precise recognition of music instruments sounds. MAIN CONCLUSION: In many sensory neural profound hearing losses as well as auditory neuropathy cases good results may be obtained by cochlear implantation. Early diagnosis of auditory neuropathy is important because of proper management. Lack of benefit from hearing aids in such cases seems to be indication to cochlear implantation. In described case cochlear implantation improved hearing perception one year after using speech processor. PMID- 16273870 TI - [Voice disorders in rheumatoid arthritis]. AB - The report presents three patients with coincidence of voice disorders and rheumatoid arthritis. Material consisted of 3 females aged 39, 48 and 74 years. Methods included phoniatric examination, videolaryngoscopy, stroboscopy and acoustic voice analysis MDVP). RESULTS: In the two youngest patients the laryngoscopic changes were located on vocal folds, mostly as yellow nodules. The appropriate mobility of both vocal folds was found. The MDVP analysis particularly revealed changes in parameters describing perturbation of amplitude in both cases. In the third case oedema in the region of the crico-arytenoid joints and vestibular folds were observed. MDVP analysis showed slight disturbances in the parameters describing frequency perturbation. PMID- 16273871 TI - [Nasopharyngeal carcinoma coexistent with lymph node tuberculosis, diagnostic difficulties--case report]. AB - Nasopharyngeal carcinoma occurs about 0.25-0.5% all malignancies in Poland. From the mids of a group more frequent are undifferentiated cancer (about 70%). The authors describe a case of undifferentiated nasopharyngeal cancer in 17-years old woman, which is connected with lymphonodular tuberculosis. They point to precise diagnostics of lymphonodular metastases, looking for primary carcinoma focus and difficulties in diagnosis. PMID- 16273872 TI - [Fibrous dysplasia of the temporal bone]. AB - Fibrous dysplasia of the temporal bone is a rare disease which may lead to progressive stenosis of the external auditory canal with conductive hearing loss. Sensorineural hearing loss has usually been attributed to involvement of the otic capsule or inner auditory canal. The case of fibrous dysplasia of the temporal bone in 11 years old girl was described. The conductive hearing loss with sensorineural component improved after surgical treatment. PMID- 16273873 TI - [The rare case of nasal glioma]. AB - The authors presented a case of nasal glioma in an adult man which had performed as nasal polyp. Histopathologic studies confirmed neuroglial tissue--Astrocytoma fibrillare. These tumor cells were immunohistochemically positive for glial fibrillary acidic protein as well as for S-100 protein and vimentin. There was no communication between the tumor and the cranial cavity on radiological examination (TC, MRI). Treatment is usually surgical. In this article authors describe a case of a nasal glioma that was removed via a lateral rhinotomy. PMID- 16273874 TI - [A neck chemodectoma--survey of literature and description of one case of malignant chemodectoma]. AB - Neck tumors are one of the most difficult diagnostic and therapeutic problems of the contemporary otolaryngology. Therapeutic success mainly depends on early and exact diagnosis. The goal of the study is to pay attention to the necessity of careful diagnosis and introducing computer tomography (CT) as a standard of neck tumors treatment procedure in order not to miss exceptionally rarely neck chemodectoma and to plan treatment process properly. CT can be a first step of vascular diagnostics what implicates safe operation course, providing effects and complications. Chemodectoma is a tumor coming from the parasympathetic part of the nervous system. The authors present a case of neck chemodectoma with malignant character in a 37-years-old woman with 10-years history of suspicion of a benign cyst. PMID- 16273875 TI - [Metastasis of rectal adenocarcinoma to the skull base and paranasal sinuses, with unusual clinical symptoms]. AB - INTRODUCTION: The isolated distant metastasis of digestive tract adenocarcinoma, to the head and neck region is very rare. Also, the VI cranial nerve paralysis, present, as an isolated symptom of the disease, in this case is very rare. MATERIAL AND METHODS: The diagnosis in 71 years female patient was based on CT, endoscopic examination and biopsy, and pathologic examination, with immunohistochemical differentiation between primary intestinal type adenocarcinoma of paranasal sinuses, and metastasis of adenocarcinoma from digestive tract. RESULTS: The immunohistochemical staining (CEA +, CK20 +, CK7 -, ChrA -, NSE -) proved the diagnosis. The colonoscopy revealed asymptomatic primary tumor in colon. The palliative Rtg-therapy was not commenced due to deterioration of general condition of patient. The patient died 2 months after diagnosis was established. CONCLUSIONS: The importance of differential diagnosis between the primary intestinal type adenocarcinoma in the upper respiratory tract and metastases of adenocarcinoma from digestive tract to head and neck region is crucial, due to entirely different type of treatment planning in both situations. PMID- 16273876 TI - [Use of the lymphoscintigraphy in detection of sentinel lymph nodes of the neck]. AB - Technique of the application of radioactive tracer to the primary tumor and lymphoscintigraphy with use of the stationary and handheld gamma probe was presented. Various types of the used radioactive tracers were characterized. Scintigraphical criteria for sentinel nodes recognition were discussed. Advantages and limitations of the use of lymphoscintigraphy for sentinel nodes detection were described. Necessity of further study to determine feasibility of sentinel nodes radiolocalization in stage N0 in head and neck cancer was pointed out. PMID- 16273877 TI - [Horizontal glottectomy--oncological and functional results. Part II. Morphology of the glottis and perceptive-acoustics characteristic of the voice and speech after horizontal glottectomy]. AB - INTRODUCTION: The authors showed findings concerning glottis morphology and perceptual-acoustic characteristics of voice and speech after partial classical (PCGLg) and extended glottic partial laryngectomy (PEGLg). MATERIAL AND METHODS: 10 patients (9 M., 1 F. average age 56 (min. 47 max. 65) were examined. All patients were undergone glottic partial laryngectomy: a) classical (n = 5) b) extended of vocal process (n = 3) with (n = 2) or without (n = 1) the removal of the mucous false folds, c) extended of part of arytenoid cartilage with (n = 2) or without (n = 1) the removal of the mucous of the false folds. The following examinations were executed: phoniatric, videolaryngoscopic and perceptual acoustic analysis. RESULTS: After PCGLg and one extended of vocal process, voice and speech has mostly characterized of features of hypofunction dysphonia. Hyperfunction was found in patients after removal of the mucous of the false folds due to leucoplakia. In case of removing of a part of arythenoid cartilage the notable or entire standstill or lack of full phonatory closure were found. The phonetical-acoustic analysis showed that in patients using melodious voice, the character of the source of actuating was periodically-noise, with the component of noise in all range of the course of the acoustic signal of voice. The parameters such as F0, jitter, shimmer does not make coherent conclusions and are less useful in the assessment of the quality of voice. CONCLUSIONS: In case of the resection of the part of the arythenoid cartilage during glottis laryngectomy, we take into account lack of full phonatory closure and using whisper by the patients. Obtaining the reliable conclusions needs continuations of the investigations and increasing number of patients. These researches are in progress. PMID- 16273880 TI - [29th Midwinter Symposium on Practical Challenges in Otolaryngology. 21 25.02.2005, Snowmass, Colorado, USA]. PMID- 16273881 TI - 'Show-stopping' disagreements may buy merger delay--Warner. PMID- 16273882 TI - DoH allows commissioning to be outsourced in Oxfordshire. PMID- 16273883 TI - Bulling. He gave me a photo of Mussolini. He was laughing. It was totally demeaning and humilitating. PMID- 16273884 TI - On more work for NICE. PMID- 16273885 TI - Data briefing. Public health:. PMID- 16273886 TI - Mental health. Stars in their eyes. AB - In 2004, Worcestershire Mental Health Partnership trust got zero stars and a damning report from the then Commission for Health Improvement. The resultant improvement strategy focused on promoting group ownership of organisational performance and ensuring that high-calibre leaders were in place to drive changes. In the final round of star-ratings, Worcestershire became the only mental health trust in the country to go from zero to three stars. PMID- 16273887 TI - On demeaning titles. PMID- 16273888 TI - Clinical management. Unknown quantities. AB - Poor performance on cancer waiting-time targets has been partly blamed on the government only recently pushing it to the top of the agenda. The first priority is to improve data collection. A number of trusts have made good progress, through streamlining services and encourging whole-trust ownership of the problem. PMID- 16273889 TI - British variations. Cure-me-quick. PMID- 16273890 TI - Topic selection. Take your pick. PMID- 16273891 TI - Roundtable discussion. Appraise be. PMID- 16273892 TI - Public health. Rhetoric into reality. PMID- 16273893 TI - Staff absence. Ill feeling. PMID- 16273894 TI - Recruitment. Not from round 'ere. PMID- 16273895 TI - Randomized, controlled study of inhaled fluticasone propionate, oral administration of prednisone, and environmental management of horses with recurrent airway obstruction. AB - OBJECTIVE: To determine whether administration of glucocorticoids provides additional benefits to environmental management of horses with recurrent airway obstruction (RAO). ANIMALS: 28 horses with RAO. PROCEDURE: Horses were classified as having mild, moderate, or severe RAO. Within each category, horses were randomly assigned to receive inhaled fluticasone propionate, inhaled control substance, or oral administration of prednisone. During the 4-week study, horses were maintained outdoors and fed a pelleted feed. Clinical scores, pulmonary function, results of cytologic examination of bronchoalveolar lavage fluid (BALF), and adrenal gland function were determined before and 2 and 4 weeks after initiation of treatment. RESULTS: Clinical score and pulmonary function of all RAO-affected horses improved during the treatment period. After 4 weeks, clinical scores and pulmonary function of horses treated with a glucocorticoid were not different from those for the control treatment. In horses with severe RAO, treatment with fluticasone for 2 weeks resulted in significantly greater improvement in pulmonary function, compared with pulmonary function after treatment with prednisone or the control substance. Treatment with a glucocorticoid for 4 weeks and a low-dust environment did not have any effect on cellular content of BALF Treatment with prednisone for 2 weeks resulted in a significant decrease in serum cortisol concentration, compared with concentrations after administration of fluticasone or the control substance. CONCLUSIONS AND CLINICAL RELEVANCE: Environmental management is the most important factor in the treatment of horses with RAO. Early treatment with inhaled fluticasone can help accelerate recovery of horses with severe RAO. PMID- 16273896 TI - Rate of change of oxygen concentration for a large animal circle anesthetic system. AB - OBJECTIVE: To describe the effects of changes in circuit volume and oxygen inflow rate on inspired oxygen concentration for a large animal circle anesthetic system. STUDY POPULATION: A large animal circle anesthetic system, a 10 L/min flowmeter, and 20- and 40-L breathing bags. PROCEDURE: Circuit volume was determined by a carbon dioxide dilution technique. Oxygen flow rates of 3, 6, and 10 L/min were delivered to the circuit with the large breathing bag, and a flow rate of 6 L/min was used with the small bag. Gas samples were collected during a 20-minute period. The time constant (tau) and half-time (T1/2) were calculated and compared with measured values. RESULTS: Mean +/- SEM volume of the breathing circuit with a 20- and 40-L breathing bag was 32.97 +/- 0.91 L and 49.26 +/- 0.58 L, respectively. The tau from measurements was 11.97, 6.10, and 3.60 minutes at oxygen flow rates of 3, 6, and 10 L/min, respectively, for the large breathing bag and 3.73 minutes at a flow rate of 6 L/min for the small breathing bag. The T1/2 was 8.29, 4.22, and 2.49 minutes at oxygen flow rates of 3, 6, and 10 L/min, respectively, for the large breathing bag and 2.58 minutes for the small breathing bag. CONCLUSIONS AND CLINICAL RELEVANCE: This study emphasizes that there are delays in the rate of increase in the inspired oxygen concentration that accompany use of conventional large animal circle anesthetic systems and low rates of inflow for fresh oxygen. PMID- 16273897 TI - Products resulting from cleavage of the interglobular domain of aggrecan in samples of synovial fluid collected from dogs with early- and late-stage osteoarthritis. AB - OBJECTIVE: To investigate interglobular domain (IGD) cleavage of aggrecan in dogs with naturally developing osteoarthritis (OA). SAMPLE POPULATION: Samples of synovial fluid (SF) obtained from 3 cubital (elbow) joints and 3 stifle joints of 4 clinically normal dogs, 24 elbow joints of 12 dogs with early-stage OA, 8 stifle joints of 5 dogs with early-stage OA, and 10 stifle joints of 9 dogs with late-stage OA. PROCEDURE: Fractions of SF were assayed for total glycosaminoglycan (GAG) content and also subjected to Western blot analysis by use of monoclonal antibodies against neoepitopes generated by cleavage of the IGD of the aggrecan protein core by matrix metalloproteinase (MMP; BC-14) and aggrecanase (BC-3). RESULTS: Total GAG content of SF from joints of clinically normal dogs did not differ from that of dogs with early-stage OA. The GAG content of SF from joints of dogs with late-stage OA was significantly lower, compared with GAG content for other SF samples. Aggrecanase-generated fragments were detected in SF from all groups but not in all samples. Matrix metalloproteinase generated fragments were not detected in any SF samples. In early-stage OA, high molecular-weight aggrecanase-generated aggrecan catabolites were evident. CONCLUSIONS AND CLINICAL RELEVANCE: GAG content of SF obtained from dogs with late-stage OA is significantly decreased, suggesting proteoglycan depletion of cartilage. Aggrecanases, but not MMPs, are the major proteolytic enzymes responsible for IGD cleavage of aggrecan in canine joints. Analyses of SF samples to detect aggrecanase-generated catabolites may provide an early biomarker for discriminating early- and late-stage OA in dogs. PMID- 16273898 TI - Development of a physiologic-based pharmacokinetic model for estimating sulfamethazine concentrations in swine and application to prediction of violative residues in edible tissues. AB - OBJECTIVE: To develop a flow-limited, physiologic-based pharmacokinetic model for use in estimating concentrations of sulfamethazine after IV administration to swine. SAMPLE POPULATION: 4 published studies provided physiologic values for organ weights, blood flows, clearance, and tissue-to-blood partition coefficients, and 3 published studies provided data on plasma and other tissue compartments for model validation. PROCEDURE: For the parent compound, the model included compartments for blood, adipose, muscle, liver, and kidney tissue with an extra compartment representing the remaining carcass. Compartments for the N acetyl metabolite included the liver and the remaining body. The model was created and optimized by use of computer software. Sensitivity analysis was completed to evaluate the importance of each constant on the whole model. The model was validated and used to estimate a withhold interval after an IV injection at a dose of 50 mg/kg. The withhold interval was compared to the interval estimated by the Food Animal Residue Avoidance Databank (FARAD). RESULTS: Specific tissue correlations for plasma, adipose, muscle, kidney, and liver tissue compartments were 0.93, 0.86, 0.99, 0.94, and 0.98, respectively. The model typically overpredicted concentrations at early time points but had excellent accuracy at later time points. The withhold interval estimated by use of the model was 120 hours, compared with 100 hours estimated by FARAD. CONCLUSIONS AND CLINICAL RELEVANCE: Use of this model enabled accurate prediction of sulfamethazine pharmacokinetics in swine and has applications for food safety and prediction of drug residues in edible tissues. PMID- 16273899 TI - Pharmacokinetics and tissue distribution of itraconazole after oral and intravenous administration to horses. AB - OBJECTIVE: To determine the pharmacokinetics of itraconazole after IV or oral administration of a solution or capsules to horses and to examine disposition of itraconazole in the interstitial fluid (ISF), aqueous humor, and polymorphonuclear leukocytes after oral administration of the solution. ANIMALS: 6 healthy horses. PROCEDURE: Horses were administered itraconazole solution (5 mg/kg) by nasogastric tube, and samples of plasma, ISF, aqueous humor, and leukocytes were obtained. Horses were then administered itraconazole capsules (5 mg/kg), and plasma was obtained. Three horses were administered itraconazole (1.5 mg/kg, IV), and plasma samples were obtained. All samples were analyzed by use of high-performance liquid chromatography. Plasma protein binding was determined. Data were analyzed by compartmental and noncompartmental pharmacokinetic methods. RESULTS: Itraconazole reached higher mean +/- SD plasma concentrations after administration of the solution (0.41 +/- 0.13 microg/mL) versus the capsules (0.15 +/- 0.12 microg/mL). Bioavailability after administration of capsules relative to solution was 33.83 +/- 33.08%. Similar to other species, itraconazole has a high volume of distribution (6.3 +/- 0.94 L/kg) and a long half-life (11.3 +/- 2.84 hours). Itraconazole was not detected in the ISF, aqueous humor, or leukocytes. Plasma protein binding was 98.81 +/- 0.17%. CONCLUSIONS AND CLINICAL RELEVANCE: Itraconazole administered orally as a solution had higher, more consistent absorption than orally administered capsules and attained plasma concentrations that are inhibitory against fungi that infect horses. Administration of itraconazole solution (5 mg/kg, PO, q 24 h) is suggested for use in clinical trials to test the efficacy of itraconazole in horses. PMID- 16273900 TI - Evaluation of analgesia resulting from extracorporeal shock wave therapy and radial pressure wave therapy in the limbs of horses and sheep. AB - OBJECTIVE: To identify the duration and potential mechanisms of analgesia following extracorporeal shock wave therapy (ESWT) and radial pressure wave therapy (RPWT) in limbs of horses and sheep. ANIMALS: 6 horses and 30 sheep. PROCEDURE: An electrical stimulus was used to identify the nociceptive threshold for each horse daily for 3 days before treatment (baseline) with ESWT or RPWT, 8 hours after treatment, and at 24-hour intervals for 7 days after treatment. Testing was conducted for the treatment field (midmetacarpus or midmetatarsus) and nerve field (medial and lateral forelimb heel bulbs) distal to a treatment site that included the nerve on the abaxial surface of the proximal sesamoid bone. All 4 limbs of 30 sheep were treated with ESWT, RPWT, or a sham treatment. Two sheep were euthanatized daily and tissue harvested for histologic evaluation of nerves, and concentrations of substance P and calcitonin gene-related peptide were measured in the skin and periosteum. RESULTS: Values did not differ significantly between baseline and after treatment for the treatment field or nerve field sensation. There was a large difference in the slope when data for horses were plotted for the first 3 days after treatment, compared with the slope for days 4 to 7 after treatment. No differences were found in neuropeptide concentrations after treatment of the sheep, but there was an inflammatory response in the treated nerves. CONCLUSIONS AND CLINICAL RELEVANCE: A small cutaneous analgesic effect may exist at the treatment site for approximately 3 days after ESWT or RPWT in horses. PMID- 16273901 TI - Vasopressin, cortisol, and catecholamine concentrations in dogs with dilated cardiomyopathy. AB - OBJECTIVE: To evaluate plasma concentrations and urinary excretion of vasopressin and cortisol and urinary excretion of catecholamines in dogs with dilated cardiomyopathy (DCM). ANIMALS: 15 dogs with clinical signs of DCM, 15 dogs with preclinical DCM, and 15 control dogs. PROCEDURE: Physical examinations, thoracic radiography, ECG, and echocardiography were performed on all dogs. Blood and urine samples were collected. RESULTS: Plasma concentration of vasopressin and the urine cortisol-to-urine creatinine ratio were significantly increased in dogs with clinical signs of DCM and dogs with preclinical DCM, compared with control dogs. Plasma vasopressin concentration was significantly higher in dogs with clinical signs of DCM, compared with dogs with preclinical DCM. Urine vasopressin to-urine creatinine ratio was significantly increased in dogs with clinical signs of DCM, compared with dogs with preclinical DCM and control dogs. Urine epinephrine-to-urine creatinine ratio and urine norepinephrine-to-urine creatinine ratio were significantly increased in dogs with clinical signs of DCM, compared with control dogs. Plasma concentration of cortisol and urine dopamine to-urine creatinine ratio did not differ significantly among groups. CONCLUSIONS AND CLINICAL RELEVANCE: According to this study, the neuroendocrine pattern is changed in dogs with preclinical DCM. These changes are even more pronounced in dogs with clinical signs of DCM. Analysis of concentrations of vasopressin, cortisol, and catecholamines may aid in identification of the clinical stages of DCM. These findings may also provide a basis for additional studies of the possible beneficial effects of vasopressin antagonists and beta-adrenergic receptor antagonists in the treatment of dogs with congestive heart failure and DCM. PMID- 16273902 TI - Serum creatine kinase response to exercise during dexamethasone-induced insulin resistance in Quarter Horses with polysaccharide storage myopathy. AB - OBJECTIVE: To determine effects of dexamethasone on insulin sensitivity, serum creatine kinase (CK) activity 4 hours after exercise, and muscle glycogen concentration in Quarter Horses with polysaccharide storage myopathy (PSSM). ANIMALS: 4 adult Quarter Horses with PSSM. PROCEDURE: A 2 x 2 crossover design was used with dexamethasone (0.08 mg/kg) or saline (0.9% NaCl) solution administered IV every 48 hours. Horses were exercised on a treadmill daily for 3 wk/treatment with a 2-week washout period between treatments. Serum CK activity was measured daily 4 hours after exercise. At the end of each treatment period, serum cortisol concentrations were measured, a hyperinsulinemic euglycemic clamp (HEC) technique was performed, and muscle glycogen content was determined. RESULTS: Mean +/- SEM serum cortisol concentration was significantly lower after 48 hours for the dexamethasone treatment (0.38 +/- 0.08 mg/dL), compared with the saline treatment (4.15 +/- 0.40 mg/dL). Dexamethasone significantly decreased the rate of glucose infusion necessary to maintain euglycemia during the HEC technique, compared with the saline treatment. Muscle glycogen concentrations and mean CK activity after exercise were not altered by dexamethasone treatment, compared with the saline treatment. CONCLUSIONS AND CLINICAL RELEVANCE: Dexamethasone significantly reduced whole-body insulin-stimulated glucose uptake in Quarter Horses with PSSM after a 3-week period but did not diminish serum CK response to exercise or muscle glycogen concentrations in these 4 horses. Therefore, a decrease in glucose uptake for 3 weeks did not appear to alleviate exertional rhabdomyolysis in these horses. It is possible that long-term treatment may yield other results. PMID- 16273903 TI - Use of corrosion casting techniques to evaluate coronary collateral vessels and anastomoses in hearts of canine cadavers. AB - OBJECTIVE: To study and investigate branching patterns of the canine coronary arteries and collateral circulation by use of corrosion casting techniques. SAMPLE POPULATION: 31 hearts obtained from cadavers of clinically normal dogs of various ages and breeds and of either sex. PROCEDURE: 3-dimensional reproduction of coronary arteries was achieved by postmortem injection and perfusion with casting materials into the aortic sinus via the ascending aorta. Perfused hearts were macerated and carefully irrigated; the air-dried specimens were examined macroscopically and with a magnifying headset. RESULTS: Collateral arteries and inter- and intra-arterial anastomoses were successfully detected in 8 corrosion cast specimens. In total, 9 coronary collateral arteries and 3 interarterial anastomoses were found. CONCLUSIONS AND CLINICAL RELEVANCE: Our finding of coronary collateral arteries in canine hearts is in agreement with recent findings in coronary flow study. On the basis of our results, vasodilation treatment to improve collateral vessel remodeling in dogs with myocardial dysfunction may be warranted. PMID- 16273905 TI - Platelet function and association of bovine viral diarrhea virus with platelets of persistently infected cattle. AB - OBJECTIVE: To determine whether viral involvement with platelets obtained from cattle persistently infected (PI) with bovine viral diarrhea virus (BVDV) is associated with altered platelet function or decreased platelet counts. SAMPLE POPULATION: Platelets obtained from 8 cattle PI with BVDV and 6 age-, sex-, and breed-matched uninfected control cattle. PROCEDURE: Manual platelet counts were determined, and platelet function was assessed through optical aggregometry by use of the aggregation agonists ADP and platelet-activating factor. Identification of BVDV in serum and preparations of purified platelets was determined by use of virus isolation tests. RESULTS: No significant difference in platelet counts was detected between cattle PI with BVDV and control cattle. In response to the aggregation agonists, maximum aggregation percentage and slope of the aggregation curve were not significantly different between cattle PI with BVDV and control cattle. We isolated BVDV from serum of all PI cattle and from purified platelets of 6 of 8 PI cattle, but BVDV was not isolated from serum or platelets of control cattle. CONCLUSIONS AND CLINICAL RELEVANCE: Isolation of BVDV from platelets in the peripheral circulation of cattle immunotolerant to BVDV does not result in altered platelet function or decreases in platelet counts. PMID- 16273906 TI - Computed tomography and cross-sectional anatomy of the brain in clinically normal dogs. AB - OBJECTIVE: To provide a detailed anatomic description of brain structures in clinically normal dogs by means of computed tomography (CT). ANIMALS: 4 clinically normal adult German Shepherd Dogs weighing 30 to 35 kg. PROCEDURE: Each dog was anesthetized and positioned in ventral recumbency for CT examination of the brain; transverse scans were completed at 2-mm intervals from the cribriform plate of the ethmoid bone to the cranial part of the atlas by use of a third-generation CT scanner. Contrast material was injected IV, and a second series of scans was completed. Images (with or without contrast) from all dogs were reviewed by use of a soft tissue setting (window width, 150 Hounsfield units; window level, 50 Hounsfield units). One of the dogs was euthanatized, and a 3.5% formaldehyde solution was perfused via the common carotid arteries. After fixation, the brain was embedded in gelatin and sectioned into 5-mm-thick transverse sections by use of a stainless-steel knife. Anatomic sections were photographed and compared with the corresponding CT views. RESULTS: Most features of the brain that were identified on anatomic sections could be identified on the corresponding CT scans despite the low contrast between structures, particularly if adjacent bony and soft tissue structures were used as landmarks. Additional anatomic structures surrounding the brain were also identifiable on the CT images. CONCLUSIONS AND CLINICAL RELEVANCE: Images obtained in this study could be used as a guide for evaluation of CT images of the brain in dogs with brain diseases. PMID- 16273904 TI - Effects of osteogenic inducers on cultures of canine mesenchymal stem cells. AB - OBJECTIVE: To examine age-related efficacy of bone morphogenetic protein (BMP)-2, ascorbate, and dexamethasone as osteogenic inducers in canine marrow-derived stromal cells (MSCs). SAMPLE POPULATION: Samples of femoral bone marrow obtained from 15 skeletally immature (< 1 year old) and 4 skeletally mature (> 1.5 years old) dogs. PROCEDURE: First-passage canine MSC cultures were treated with 100 microg of ascorbate phosphate/mL, 10(-7)M dexamethasone, 100 ng of BMP-2/mL, or a combination of these osteoinducers. On day 6, cultures were harvested for quantitation of alkaline phosphatase (ALP) activity and isolation of RNA to prepare cDNA for real-time polymerase chain reaction analyses of osteoblast markers. RESULTS: Early markers of osteogenesis were induced in canine MSCs by BMP-2 but not dexamethasone. In young dogs, the combination of BMP-2 and ascorbate yielded the highest ALP mRNA concentrations and activity. This combination also induced significant increases in mRNA for osteopontin and runt domain transcription factor 2. In comparison to MSCs from immature dogs, those from mature dogs had diminished ALP activity in response to BMP and ascorbate. Results for cultures treated with 3,4-dehydroproline suggested that ascorbate induced production of extracellular matrix was important for maximal BMP-2 response in canine MSCs. CONCLUSIONS AND CLINICAL RELEVANCE: BMP-2 was capable of inducing markers of osteogenesis in short-term cultures of canine MSCs. In MSCs obtained from skeletally immature dogs, ascorbate was required for maximal effects of BMP These results define optimal conditions for stem cell osteogenesis in dogs and will facilitate development of stem cell-based treatments for dogs with fractures. PMID- 16273907 TI - Effects of radial shock waves on membrane permeability and viability of chondrocytes and structure of articular cartilage in equine cartilage explants. AB - OBJECTIVE: To investigate in vitro effects of radial shock waves on membrane permeability, viability, and structure of chondrocytes and articular cartilage. SAMPLE POPULATION: Cartilage explants obtained from the third metacarpal and metatarsal bones of 6 horses. PROCEDURE: Equine cartilage was subjected to radial shock waves and then maintained as explants in culture for 48 hours. Treatment groups consisted of a negative control group; application of 500, 2,000, and 4,000 impulses by use of a convex handpiece (group A); and application of 500, 2,000, and 4,000 impulses by use of a concave handpiece (group B). Effects on explant structure were evaluated by use of environmental scanning electron microscopy (ESEM). Membrane permeability was determined by release of lactate dehydrogenase (LDH). Chondrocyte viability was assessed by use of vital cell staining. Comparisons of LDH activity and nonviable cell percentages were performed by ANOVA. RESULTS: Cell membrane permeability increased significantly after application of 2,000 and 4,000 impulses in groups A and B. A significant decrease in cell viability was observed for application of 4,000 impulses in explants of group A. There was no detectable damage to integrity of cartilage explants observed in any treatment group by use of ESEM. CONCLUSIONS AND CLINICAL RELEVANCE: Radial shock waves do not appear to structurally damage articular cartilage but do impact chondrocyte viability and membrane permeability. Caution should be exercised when extremely high periarticular pulse doses are used until additional studies can determine the long-term outcome of these effects and appropriate periarticular treatment regimens can be validated. PMID- 16273908 TI - Evaluation of papillomaviruses associated with cyclosporine-induced hyperplastic verrucous lesions in dogs. AB - OBJECTIVE: To determine whether cyclosporine A-induced hyperplastic skin lesions of dogs were associated with papillomavirus infections. ANIMALS: 9 dogs that were treated with cyclosporine A and developed hyperplastic skin lesions. PROCEDURE: History and clinical and histopathologic data were collected. Paraffin-embedded skin biopsy specimens from hyperplastic skin lesions were immunostained for common papillomavirus genus-specific structural antigens by use of a polyclonal rabbit anti-bovine papillomavirus type 1 antiserum. Sections from each tissue block underwent DNA extraction, and polymerase chain reaction (PCR) assays were performed with several sets of primers to amplify a wide range of papillomavirus DNA from humans and other animals. RESULTS: In 7 of 9 dogs, there were more than 10 hyperplastic skin lesions that microscopically resembled those of psoriasiform lichenoid dermatosis. In those dogs, results of testing for papillomavirus via immunohistochemical analyses and PCR assays were negative. In the other 2 dogs, there were only 1 and 3 verrucous lesions, and in those dogs, histologic evaluation revealed koilocytes and nuclear viral inclusions that were immunoreactive for papillomavirus antigens. Papillomavirus DNA was amplified from both dogs. One of the sequences was characteristic for the canine oral papillomavirus, whereas the other had similarities with the recently described canine papillomavirus 2. CONCLUSIONS AND CLINICAL RELEVANCE: In dogs, hyperplastic skin lesions occasionally develop during treatment with cyclosporine A. Most of the lesions resemble those of psoriasiform lichenoid dermatosis, although papillomavirus can be detected in some instances. PMID- 16273909 TI - Evaluation of the concentration of marbofloxacin in alveolar macrophages and pulmonary epithelial lining fluid after administration in dogs. AB - OBJECTIVE: To determine concentrations of marbofloxacin in alveolar macrophages (AMs) and epithelial lining fluid (ELF) and compare those concentrations with plasma concentrations in healthy dogs. ANIMALS: 12 adult mixed-breed and purebred hounds. PROCEDURE: 10 dogs received orally administered marbofloxacin at a dosage of 2.75 mg/kg every 24 hours for 5 days. Two dogs served as nontreated controls. Fiberoptic bronchoscopy and bronchoalveolar lavage procedures were performed while dogs were anesthetized with propofol, approximately 6 hours after the fifth dose. The concentrations of marbofloxacin in plasma and bronchoalveolar fluid (cell and supernatant fractions) were determined by use of high-performance liquid chromatography with detection of fluorescence. RESULTS: Mean +/- SD plasma marbofloxacin concentrations 2 and 6 hours after the fifth dose were 2.36 +/- 0.52 microg/mL and 1.81 +/- 0.21 microg/mL, respectively. Mean +/- SD marbofloxacin concentration 6 hours after the fifth dose in AMs (37.43 +/- 24.61 microg/mL) was significantly greater than that in plasma (1.81 +/- 0.21 microg/mL) and ELF (0.82 +/- 0.34 microg/mL), resulting in a mean AM concentration-to-plasma concentration ratio of 20.4, a mean AM:ELF ratio of 60.8, and a mean ELF-to-plasma ratio of 0.46. Marbofloxacin was not detected in any samples from control dogs. CONCLUSIONS AND CLINICAL RELEVANCE: Marbofloxacin concentrations in AMs were greater than the mean inhibitory concentrations of major bacterial pathogens in dogs. Results indicated that marbofloxacin accumulates in AMs at concentrations exceeding those reached in plasma and ELF The accumulation of marbofloxacin in AMs may facilitate treatment for susceptible intracellular pathogens or infections associated with pulmonary macrophage infiltration. PMID- 16273910 TI - Evaluation of intra- and interobserver reliability and image reproducibility to assess usefulness of high-resolution ultrasonography for measurement of anterior segment structures of canine eyes. AB - OBJECTIVE: To evaluate the usefulness of high-resolution ultrasonography (HRUS) for measurements of anterior segment structures in canine eyes. ANIMALS: 4 clinically normal Beagles. PROCEDURE: Images were obtained from 8 eyes with a handheld 20-MHz transducer. Eleven anterior segment structures on each image were measured 5 times by 2 independent observers. Coefficients of variation (CVs) for measurements were used to assess intraobserver reliability. Interobserver reliability was assessed by comparing measurements obtained by the 2 observers from the same images. Five images were sequentially obtained from 2 locations (ie, superior and temporal) to evaluate image reproducibility. Anterior segment structures were measured once on each image; image reproducibility was assessed by use of the CV for each parameter measured. Imaging location was assessed by comparison of CV for measurements from each location. RESULTS: CVs were < 10% for observer A for all measurements except the ciliary cleft area (11.63%). The CVs were > 10% for observer B for measurements of the angle recess area (18.51%) and ciliary cleft width (17.44%) and area (16.01%). Significant differences in measurements between observers were found for 5 of 11 anterior segment structures. Imaging the superior aspect of the globe provided the most reproducible images, although image reproducibility was still somewhat variable, with the highest and lowest CVs for measurements of 33.01% and 11.32%, respectively, in the superior position. CONCLUSIONS AND CLINICAL RELEVANCE: High resolution ultrasound images can be used to reliably measure various anterior segment structures. Clinically relevant findings in the anterior segment of canine eyes may be detectable by use of HRUS. PMID- 16273911 TI - Serologic responses of dogs given a commercial vaccine against Leptospira interrogans serovar pomona and Leptospira kirschneri serovar grippotyphosa. AB - OBJECTIVE: To evaluate serum titers obtained by use of the microscopic agglutination test (ie, MAT titers) to Leptospira interrogans serovar pomona and autumnalis and Leptospira kirschneri serovar grippotyphosa in dogs given a commercial vaccine against serovars pomona and grippotyphosa. ANIMALS: Forty 12 week-old puppies and 20 mature Beagles. PROCEDURE: Puppies received a commercial vaccine against serovars pomona and grippotyphosa at 12 weeks of age, then received a booster vaccine and 3 weeks later; mature dogs received the vaccine once. Serum MAT titers to serovars pomona, autumnalis, and grippotyphosa were measured before vaccination and at 2, 4, 6, 10, and 16 weeks after the first or only vaccination. RESULTS: Of the 40 puppies vaccinated, 40, 0, and 40 developed MAT titers of > 100 after vaccination to serovars pomona, grippotyphosa, and autumnalis, respectively. Microscopic agglutination test titers to serovar autumnalis were higher than MAT titers to serovars pomona and grippotyphosa and persisted in some dogs for 16 weeks (6 weeks longer than for titers to serovar pomona). Of the 20 mature dogs, 13, 5, and 20 developed MAT titers of > 100 at 2 weeks to serovars pomona, grippotyphosa, and autumnalis, respectively. Titers to serovar pomona were higher and persisted in some dogs beyond 16 weeks after vaccination, compared with titers to serovars pomona and grippotyphosa, which persisted for 10 and 6 weeks, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Subunit vaccines against serovars pomona and grippotyphosa induce MAT titers not only to homologous antigens but also to serovar autumnalis, which could lead to a misdiagnosis of leptospirosis caused by serovar autumnalis. PMID- 16273912 TI - Characterization of protection from systemic infection and disease by use of a modified-live noncytopathic bovine viral diarrhea virus type 1 vaccine in experimentally infected calves. AB - OBJECTIVE: To evaluate protection against systemic infection and clinical disease provided by use of a modified-live noncytopathic bovine viral diarrhea virus (BVDV) type 1 vaccine in calves challenged with NY-1 BVDV. ANIMALS: 10 calves, 5 to 7 months of age. PROCEDURES: Calves were allocated (n = 5/group) to be nonvaccinated or vaccinated SC on day 0 with BVDV type 1 (WRL strain). Calves in both groups were challenged intranasally with NY-1 BVDV on day 21. Calves' rectal temperatures and clinical signs of disease were recorded daily, total and differential WBC and platelet counts were performed, and serum neutralizing antibody titers against NY-1 BVDV were determined. Histologic examinations and immunohistochemical analyses to detect gross lesions and distribution of viral antigens, respectively, were performed. RESULTS: After challenge exposure to NY-1 BVDV, nonvaccinated calves developed high rectal temperatures, increased respiratory rates, viremia, leukopenia, lymphopenia, and infection of the thymus. Vaccinated calves did not develop high rectal temperatures or clinical signs of respiratory tract disease. Vaccinated calves appeared to be protected against systemic replication of virus in that they did not develop leukopenia, lymphopenia, viremia, or infection of target organs, and infectious virus was not detected in peripheral blood mononuclear cells or the thymus. CONCLUSIONS AND CLINICAL RELEVANCE: The modified-live BVDV vaccine protected calves against systemic infection and disease after experimental challenge exposure with NY-1 BVDV. The vaccine protected calves against infection and viremia and prevented infection of target lymphoid cells. PMID- 16273913 TI - Temporality of early-term abortions associated with mare reproductive loss syndrome in horses. AB - OBJECTIVE: To characterize the temporality of dates of breeding and abortion classified as mare reproductive loss syndrome (MRLS) among mares with abortions during early gestation. ANIMALS: 2,314 mares confirmed pregnant at approximately 28 days after breeding from 36 farms in central Kentucky, including 515 mares that had early-term abortions. PROCEDURE: Farm veterinarians and managers were interviewed to obtain data for each mare that was known to be pregnant to determine pregnancy status, breeding date, last date known to be pregnant, and date of abortion. RESULTS: Mares bred prior to April 1, 2001, appeared to be at greatest risk of early-term abortion, both among and within individual farms. Mares bred in mid-February appeared to be at greatest risk of abortion, with an estimated weekly incidence rate of abortion of 66% (95% CI, 52% to 80%). CONCLUSIONS AND CLINICAL RELEVANCE: Mares in central Kentucky bred between mid February and early March were observed to be at greatest risk of early-term abortion, and risk gradually decreased to a background incidence of abortion of approximately 11%. Mares bred after April 1, 2001, appeared to be at markedly less risk, indicating that exposure to the cause of MRLS likely occurred prior to this date. PMID- 16273914 TI - Assessment of the dark-adaptation time required for recovery of electroretinographic responses in dogs after fundus photography and indirect ophthalmoscopy. AB - OBJECTIVE: To investigate the duration of dark-adaptation time required for recovery of electroretinographic responses after fundus photography or indirect ophthalmoscopy in dogs. ANIMALS: 6 dogs. PROCEDURE: Initially, scotopic-intensity series of electroretinograms (ERGs) were recorded after 20 minutes of dark adaptation. The fundus of the left eye of each dog was photographed (n = 10) or examined via indirect ophthalmoscopy for 5 minutes with moderate- (117 candela [cd]/m2) or bright-intensity (1,693 cd/m2) light; ERGs were repeated after a further 20 or 60 minutes of dark adaptation (6 procedures/dog). RESULTS: Following 20 minutes of dark adaptation after fundus photography, the b- and a wave amplitudes were reduced in response to brighter stimuli, compared with pretest ERGs; after 60 minutes of dark adaptation, ERG amplitudes had recovered. Following 20 minutes of dark adaptation after indirect ophthalmoscopy (moderate intensity light), significantly lower b-wave amplitudes were recorded in response to 2 of the brighter flash stimuli, compared with pretest ERGs; after 60 minutes of dark adaptation, ERG amplitudes had recovered. Following 20 minutes of dark adaptation after indirect ophthalmoscopy (bright-intensity light), all ERG amplitudes were significantly decreased and implicit times were significantly decreased at several flash intensities, compared with pretest ERGs; after 60 minutes of dark adaptation, ERG amplitudes and implicit times had returned to initial values, except for b-wave amplitudes recorded in response to dimmer stimuli. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that at least 60 minutes of dark adaptation should be allowed before ERGs are performed in dogs after fundus photography or indirect ophthalmoscopy. PMID- 16273915 TI - Association between subjective lameness grade and kinetic gait parameters in horses with experimentally induced forelimb lameness. AB - OBJECTIVE: To evaluate the association between subjective lameness grades and kinetic gait parameters and assess the variability in kinetic parameters in horses with experimentally induced forelimb lameness. ANIMALS: 32 horses. PROCEDURES: Forelimb lameness was induced in each horse via injection of lipopolysaccharide into 1 metacarpophalangeal joint (40 experimental trials). Subjective lameness grading and 13 kinetic gait parameters (force plate analysis) were assessed before (baseline) and at 12, 18, and 24 hours after lipopolysaccharide injection. While horses were trotting, kinetic gait analysis was performed for 8 valid repetitions at each time point. Repeated-measures analyses were performed with 8 repetitions for each kinetic parameter as the outcome, and lameness grades, time points after lipopolysaccharide injection, and repetition order as explanatory variables. Sensitivity and specificity of kinetic parameters for classification of horses as sound or lame (in relation to subjective lameness scores) were calculated. Between- and within-horse variabilities of the 13 kinetic parameters were assessed by calculation of coefficients of variation. RESULTS: Subjective lameness grades were significantly associated with most of the kinetic parameters. Vertical force peak and impulse had the lowest between- and within-horse coefficients of variation and the highest correlations with subjective lameness grade. Vertical force peak had the highest sensitivity and specificity for lameness classification. Vertical force peak and impulse were significantly decreased even in horses with mild or unobservable lameness. CONCLUSIONS AND CLINICAL RELEVANCE: Among the kinetic gait parameters, vertical force peak and impulse had the best potential to reflect lameness severity and identify subclinical forelimb gait abnormalities. PMID- 16273917 TI - Evaluation of subchondral bone mineral density associated with articular cartilage structure and integrity in healthy equine joints with different functional demands. AB - OBJECTIVE: To determine and correlate subchondral bone mineral density and overlying cartilage structure and tensile integrity in mature healthy equine stifle (low magnitude loading) and metacarpophalangeal (high magnitude loading) joints. ANIMALS: 8 healthy horses, 2 to 3 years of age. PROCEDURE: Osteochondral samples were acquired from the medial femoral condyle (FC) and medial trochlear ridge (TR) of the stifle joint and from the dorsal (MC3D) and palmar (MC3P) aspects of the distal medial third metacarpal condyles of the metacarpophalangeal joint. Articular cartilage surface fibrillation (evaluated via India ink staining) and tensile biomechanical properties were determined. The volumetric bone mineral density (vBMD) of the underlying subchondral plate was assessed via dual-energy x-ray absorptiometry. RESULTS: Cartilage staining (fibrillation), tensile moduli, tensile strength, and vBMD were greater in the MC3D and MC3P locations, compared with the FC and TR locations, whereas tensile strain at failure was less in MC3D and MC3P locations than FC and TR locations. Cartilage tensile moduli correlated positively with vBMD, whereas cartilage staining and tensile strain at failure correlated negatively with vBMD. CONCLUSIONS AND CLINICAL RELEVANCE: In areas of high joint loading, the subchondral bone had high vBMD and the articular cartilage surface layer had high tensile stiffness but signs of structural wear (fibrillation and low failure strain). The site dependent variations and relationships in this study support the concept that articular cartilage and subchondral bone normally adapt to physiologic loading in a coordinated way. PMID- 16273916 TI - Experimental canine leptospirosis caused by Leptospira interrogans serovars pomona and bratislava. AB - OBJECTIVE: To evaluate gross, histopathologic, and serum biochemical findings caused by Leptospira interrogans serovars pomona and bratislava inoculated in dogs. ANIMALS: Twenty-seven 8-week-old female Beagles. PROCEDURE: Dogs were randomly assigned to challenge or control groups. Challenge groups were conjunctivally inoculated on 3 successive days with 5 x 10(7) L interrogans serovar pomona (n = 12) or serovar bratislava (11). Clinical signs were recorded throughout the experiment, and clinical pathology assays, bacteriologic culture, and necropsies (6 or 7 dogs necropsied at each time point) were done on postinoculation day (PID) 7, 10, 14, and 20. RESULTS: Infection could not be confirmed in any serovar bratislava-inoculated dog, and control dogs remained healthy throughout the experiment. Positive culture and fluorescent antibody test results were confirmed in 11 of 12 serovar pomona-inoculated dogs. Fever and lethargy starting at PID 7 were the most common clinical signs in serovar pomona infected dogs. On day 10, gross lesions included multifocal renal and pulmonary hemorrhage and perirenal edema. Serovar pomona-inoculated dogs had histopathologic lesions including hepatitis, interstitial nephritis, and pneumonia at PID 7, 10, 14, and 20. Increases in BUN, anion gap, and bilirubin concentration occurred on PID 10, 14, and 20. Platelet counts in dogs with positive results of bacteriologic culture were decreased from baseline values on PID 10, 12, and 14. CONCLUSIONS AND CLINICAL RELEVANCE: Conjunctival inoculation with L interrogans serovar pomona resulted in a high rate of infection with concomitant hemorrhagic and inflammatory lesions of the kidneys, liver, and lungs. PMID- 16273918 TI - Measurement of plasma chromogranin A concentrations for assessment of stress responses in dogs with insulin-induced hypoglycemia. AB - OBJECTIVE: To determine whether cross-reactivity exists between canine chromogranin A (CgA) and anti-human CgA antibody and investigate the usefulness of plasma CgA concentration measurements as an index of acute stress responses in dogs. ANIMALS: 12 healthy Beagles. PROCEDURE: Canine CgA was extracted and purified from canine adrenal glands of cadaver dogs for studying cross-reactivity with anti-human CgA antibody. Western blotting with anti-human CgA antibody was performed. Blood samples were collected from dogs at 0, 10, 20, 30, 40, 60, 120, and 180 minutes after IV administration of saline (0.9% NaCl) solution or insulin. Canine plasma CgA concentrations were determined by use of a CgA ELISA kit with rabbit antiserum against the carboxy-terminal fragment of human CgA. Plasma cortisol and catecholamine (ie, norepinephrine and epinephrine) concentrations were measured by use of an ELISA and a high-performance liquid chromatography method, respectively. RESULTS: Purified canine CgA was specifically detected by use of western blot analysis and an ELISA with anti human CgA antibody. An increase in plasma CgA concentrations was observed in insulin-induced hypoglycemic dogs. Changes in plasma CgA concentration were correlated with changes in plasma cortisol or catecholamine concentrations of hypoglycemic dogs. CONCLUSIONS AND CLINICAL RELEVANCE: Use of the CgA ELISA kit for determination of human plasma CgA concentrations is applicable to the measurement of canine plasma CgA concentrations. Canine plasma CgA concentrations, along with measurements of plasma cortisol and catecholamine concentrations, correctly reflect insulin-induced hypoglycemic stressed conditions in dogs. Measurement of canine plasma CgA concentrations may provide a useful index for evaluation of an acute stress response. PMID- 16273920 TI - [Topical issues of informatization in public health and medical science]. AB - Relevant trends in informatization progress in Russian public health and medical science are justified. Actual processes in population health are considered. PMID- 16273919 TI - Assessment of scintigraphic and thermographic changes after focused extracorporeal shock wave therapy on the origin of the suspensory ligament and the fourth metatarsal bone in horses without lameness. AB - OBJECTIVE: To monitor the effect of focused extra-corporeal shock wave therapy (ESWT) on bone and bone-tendon junction of horses without lameness by use of nuclear scintigraphy and thermography. ANIMALS: 6 warmblood horses without lameness. PROCEDURE: The origin of the suspensory ligament at the metacarpus (OSL MC) and the fourth metatarsal bone were treated at 2 time points (days 0 and 16) with 2,000 shocks applied by a focused ESWT device at an energy flux density of 0.15 mJ/mm2. One forelimb and 1 hind limb were treated, and the contralateral limbs served as controls. To document the effect of focused ESWT, nuclear scintigraphy was performed on days -1, 3, 16 (before second ESWT), and 19. Thermography was performed on days -1, 0 (1 hour after first ESWT), 1, 3, 8, 16 (twice; before and 1 hour after second ESWT), and 19. On days 3, 16 (first scans), and 19, thermography was performed before scintigraphy. RESULTS: Scintigraphically, significant variations in radiopharmaceutical activity at the OSL-MC were detected in treatment and control limbs. No significant differences, however, in mean temperature or radiopharmaceutical activity could be detected by use of thermography or nuclear scintigraphy, respectively, between the treatment and control limbs at any time point in response to ESWT. CONCLUSIONS AND CLINICAL RELEVANCE: After 2 treatments of focused ESWT, no physiologic effect on the studied structures could be demonstrated by use of nuclear scintigraphy or thermography. Results of this study indicate that at currently used ESWT settings, no damage to the bone or bone-tendon junction should occur. PMID- 16273921 TI - [National program of poverty elimination and health of economically active population in Russia]. AB - Effective implementation of actual tasks on social economic development of Russia is certainly determined by significant increasing of health level of laboring people. Death rate of economically active population is an appropriate and highly informative indicator of both population health and society social well-being. Main trend in implementation of program of poverty elimination and in enhancement of national competitive capacity is effective carrying out of public program? Health of laboring population in Russia: 2004 - 2015? with highest possible employers' resource support. PMID- 16273922 TI - [Characteristics of patient informed consent in physician practice]. AB - Informed consent as one of major problems of contemporary medical law and medical ethics is considered. Basic elements of this process are revealed. Standards determining patient's competence are described. The following issues are discussed: scope of information to be presented to patient; possible alternatives to particular treatment; risk degree, specifics and purpose of proposed treatment. Peculiarities of introducing principle of informed consent into pediatric practice are emphasized. Any child is most vulnerable patient therefore additional efforts are needed to protect his rights. Necessity to obtain child's approval to be examined and to be treated is issue of pressing importance. PMID- 16273923 TI - [Characteristics of male morbidity in complicated involution period]. PMID- 16273924 TI - [Prospects of medical sanitary care development in rural areas of Republic of Tajikistan]. AB - In present social economic conditions of Republic of Tajikistan, health sector reform is carried out on basis of National concept of public health reformation. Its major elements are medical preventive institutions. They implement primary medical sanitary care in rural areas where most of country population is dwelling. Amelioration of corresponding material and technical background and enhancement of management mode aligned to WHO directives is intended. PMID- 16273925 TI - [Mortality of population living in areas of local earth's crust ruptures]. AB - In areas of local earth's crust ruptures on territory of City of Surgut (Khanty Mansy autonomous Okrug), relationship between death cases of cardiovascular diseases and malignant neoplasms and physical factors anomalies (geomagnetic field, gamma rays, radon escape) has been studied. Findings analysis demonstrated that among city residents death rates of major cardiovascular diseases, lung cancer and stomach cancer reliably prevailed in areas of local earth's crust ruptures as compared with the rest of city territory. Planning of medical care and implementing of treatment and prevention activities requires to consider dwelling in abnormal physical fields zones. PMID- 16273926 TI - [Scientific rationale of capacity requirements in emergency and acute care of urban population]. AB - Statistical data related to analysis of emergency care in Russian Federation and its subjects is presented. By the example of city of Tver calculation data on requirements in emergency and acute medical care to urban population is given. Capacity of acute medical care is determined including rate of emergency calls which, depending of reception time, can be transfered to general practitioner. PMID- 16273927 TI - [New approaches to medical social problems in caring family reproductive health]. PMID- 16273928 TI - [Medical social conditions in motivating enhancement of out-patient health care in municipal and non-governmental sectors]. PMID- 16273929 TI - [Patients opinions on in-patient substituting technologies]. PMID- 16273931 TI - [Functioning of out-patient institutions in conditions of budget gap]. PMID- 16273930 TI - [Clinical examination of children in municipal pediatric polyclinics]. AB - In 3 municipal polyclinics in city of Makhachkala, Republic of Dagestan, special research was carried out to elaborate technical guidelines enhancing quality of children clinical examinations. Quality of children clinical examinations had been assessed according to expert evaluation of 437 outpatient records. Expert appraisal revealed that in municipal pediatric polyclinics quality of children medical examinations didn't correspond the needed level. Irregular medical supervision of children by district pediatricians (23.1% of cases), district nurses (38%) and clinical specialists (68.3%) is established. It is noted that in 19.9% of cases untimely medical examinations were held. Moreover, for every forth patient the medical examination was incomplete. Health promoting activities (17.5%) and medical rehabilitation (24.8%) were carried out inadequately. On basis of research data system of actions enhancing quality of children medical examinations under municipal pediatric polyclinics practice is elaborated. PMID- 16273932 TI - [Logbook system application in regional multi-field hospital]. PMID- 16273933 TI - [On appropriateness of development of in-patient substituting technologies in diagnostic centers]. PMID- 16273934 TI - [Sanitary epidemiologic service of land forces (1918-1919)]. PMID- 16273935 TI - [Free economic society activities in field of medicine (1765-1917)]. PMID- 16273936 TI - [Major stages of homoeopathy development in Russia]. PMID- 16273937 TI - [Evolution of pediatric surgery in Vilnyus (on the occasion of centenary of first pediatric surgery hospital in Russian Empire)]. PMID- 16273938 TI - [Insurance medicine in Western Belorussia during interwar period (1921-1939)]. PMID- 16273939 TI - Pancreatitis: an introduction to history, etiology and pathogenesis. PMID- 16273940 TI - Acute pancreatitis. AB - Acute pancreatitis is characterized by the occurrence of necroinflammatory changes in the pancreas. Three types of necrosis may be distinguished: (1) interstitial tissue necrosis, which subsequently may also involve acinar and ductal cells, (2) ductal necrosis, and (3) acinar necrosis. The first type of necrosis is autodigestive in nature and is typical of the most common forms of acute pancreatitis, which are associated with alcohol, bile duct disease, metabolic conditions, and other rare factors. Clinically, these types of pancreatitis may be either mild or severe (Atlanta classification). The mild form is also known as edematous pancreatitis, because there is edematous swelling of the pancreas combined with tiny foci of interstitial (fat) necrosis. Severe or necrotizing pancreatitis shows large areas of often hemorrhagic necrosis of the pancreatic and particularly the peripancreatic tissue. The ductal type of necrosis is rare and may be seen in pancreatitis associated with prolonged circulatory failure. The acinar type of necrosis is caused by infectious agents. Complications of acute pancreatitis, such as pseudocyst, bleeding, and infection, determine the course of the disease. PMID- 16273941 TI - Chronic pancreatitis of alcoholic and nonalcoholic origin. AB - Fibroinflammatory changes to the pancreatic tissue characterize chronic pancreatitis. This article summarizes the current state of knowledge on the pathology and pathogenesis of chronic pancreatitis associated with alcohol, hereditary factors, metabolic conditions, and anatomical abnormalities. Specifically, the pathogenetic mechanisms that lead to chronic pancreatitis in patients with alcohol abuse will be discussed. In addition, brief descriptions of the features of chronic pancreatitis of nonalcoholic origin and of the pancreatic fibrosis that is not associated with symptoms of chronic pancreatitis will be given. PMID- 16273942 TI - Lymphoplasmacytic sclerosing (autoimmune) pancreatitis. AB - Lymphoplasmacytic sclerosing pancreatitis (LPSP), also known as autoimmune pancreatitis or nonalcoholic, duct destructive chronic pancreatitis, has been increasingly recognized in the past decade as a histologically distinctive type of pancreatitis that affects middle-aged patients who lack the typical risk factors for chronic pancreatitis (alcohol abuse in particular). LPSP is sometimes associated with other autoimmune diseases or fibroinflammatory lesions, although in some patients, pancreatic and biliary involvement represent the only known disease process. Many patients present with pancreatic masses clinically and radiographically simulating pancreatic carcinoma, and associated bile duct strictures enhance the resemblance. Elevated serum IgG4 levels have been described in patients with LPSP and have been used to distinguish LPSP from pancreatic carcinoma preoperatively. Although there is some heterogeneity of pathologic findings, resected cases of LPSP typically demonstrate dense periductal lymphoplasmacytic inflammation, periductal and parenchymal fibrosis, and obliterative venulitis; neutrophilic infiltration of the ductal epithelium ("granulocytic epithelial lesions") may also occur. Large tumor-like masses of fibroinflammatory tissue ("reactive fibroinflammatory pseudotumors") may develop and extend beyond the pancreas. Following surgical resection, a few patients suffer recurrence of fibroinflammatory lesions in the pancreatobiliary tree, or they may develop other manifestations of autoimmune disease elsewhere in the body. However, the overall prognosis is excellent. Response to steroid therapy has been noted. Current studies are focusing on identifying additional preoperative diagnostic tests and on characterizing possible variants of LPSP. This review presents the defining clinical and pathologic features of LPSP and discusses the ongoing efforts to understand the pathogenesis of this disease. PMID- 16273943 TI - Paraduodenal pancreatitis: a clinico-pathologically distinct entity unifying "cystic dystrophy of heterotopic pancreas", "para-duodenal wall cyst", and "groove pancreatitis". AB - A distinct form of chronic pancreatitis occurring predominantly in and around the duodenal wall (near the minor papilla) has been reported under various names, including cystic dystrophy of heterotopic pancreas, pancreatic hamartoma of duodenum, para-duodenal wall cyst, myoadenomatosis, and groove pancreatitis. Our experience with these lesions and the review of the literature show that these lesions have the following common characteristics: (1) The duodenal wall contains dilated ducts, some with inspissated secretions, and pseudocystic changes as well as adjacent stromal reactions including hypercellular granulation tissue, foreign body type giant cell reaction engulfing mucoprotein material, and myofibroblastic proliferation. (2) Brunner's gland hyperplasia is typically present. (3) Dense myoid stromal proliferation, with intervening rounded lobules of pancreatic acinar tissue, creates a histologic picture reminiscent of "myoadenomatosis," "pancreatic hamartoma," or even leiomyoma in some cases. (4) Spillover of fibrosis into the adjacent pancreas and soft tissue occurs, especially in the "groove" area (between the pancreas, common bile duct and duodenum), including the region around the common bile duct. (5) Clinically, these lesions often mimic "pancreas cancer" or periampullary tumors, because of marked scarring as well as the ill-defined borders of the process. Patients with these findings are predominantly males, 40-50 years old, with a history of alcohol abuse. That the process is often centered in the region of minor papilla (and the adjacent pancreas) suggests that an anatomic variation of the ductal system may render this area particularly susceptible to the effects of alcoholic injury, and the myo-adenomatoid and cystic changes on the duodenal wall may in turn represent changes related to a localized recurrent pancreatitis. In conclusion, these clinicopathologic findings characterize a distinctive process that can be referred to as paraduodenal pancreatitis. PMID- 16273944 TI - The inflamed pancreas transplant: histological differential diagnosis. AB - Excellent results are obtained with pancreas transplantation for the treatment of diabetes mellitus. The clinical parameters indicating graft dysfunction are nonspecific, and evaluation of a graft biopsy is often necessary to determine the etiology of graft dysfunction and assess the status of the graft. Inflammation of the pancreas allograft is the most common histological feature seen in core biopsies. In most cases, the inflammation is secondary to acute rejection, chronic rejection, and infectious complications. Although in many ways the morphological features of these processes overlap, there are also important differences that can help the pathologist to reach the correct diagnosis. PMID- 16273945 TI - Pancreatic pseudotumors: non-neoplastic solid lesions of the pancreas that clinically mimic pancreas cancer. AB - In the pancreas, a variety of non-neoplastic conditions may form solid masses that may mimic cancer. Up to 5% of pancreatectomies performed with the preoperative clinical diagnosis of carcinoma will prove to be non-neoplastic by pathologic examination, although this figure is decreasing with improved diagnostic modalities. Chronic inflammatory lesions are the leading cause of this phenomenon ("pseudotumoral pancreatitis"), and among these, autoimmune and paraduodenal pancreatitides (discussed separately in this issue) are most important. In this article, we will focus on the noninflammatory lesions that may form tumor-like lesions of the pancreas. Adenomyomatous hyperplasia of ampulla of Vater is a subtle lesion that is difficult to define; larger examples (>5 mm) have been found to be the cause of obstructive jaundice. Accessory (heterotopic) spleen may form a well-defined nodule within the tail of the pancreas and is typically mistaken for endocrine neoplasm. Lipomatous hypertrophy is the replacement of pancreatic tissue with mature adipose tissue that occasionally leads to moderate to marked enlargement of the pancreas. Hamartomas are very rare if the entity is defined strictly. They are characterized by irregularly arranged mature pancreatic elements admixed with stromal tissue. A cellular, spindle-cell variant with c-kit (CD117) expression is recognized. Pseudolymphoma forms well defined nodules composed of hyperplastic lymphoid tissue. Rarely, foreign-body deposits, granulomatous inflammations (such as sarcoidosis or tuberculosis), and congenital lesions may form tumoral lesions. In conclusion, it is important to recognize the types of conditions that form pseudotumors in the pancreas so that they can be distinguished from ductal adenocarcinomas, especially clinically, but also pathologically. Nonspecific terms such as "inflammatory pseudotumor" ought to be avoided, and every attempt should be made to classify a "pseudotumor" into a more specific diagnostic category discussed above. PMID- 16273947 TI - Serving the unserved: informal refuse collection in Mexico. AB - The incomplete collection of municipal solid waste is a significant problem in the cities of many developing countries. Mexican cities collect less than 75% of the waste generated. The uncollected waste generates pollution and poses risks to human health and the environment. Low-income communities comprise most areas that lack refuse collection. In many of the areas where there is no official collection service, informal collectors provide this service for a fee. This paper analyses informal refuse collection in three Mexican cities, and argues that it can supplement municipal collection, create jobs, benefit the economy and help clean up the urban environment. PMID- 16273946 TI - Chronic pancreatitis or pancreatic ductal adenocarcinoma? AB - The histopathologic distinction of ductal adenocarcinoma (DA) of the pancreas from chronic pancreatitis (CP) is a well-known challenge. Several parameters have been determined by the authors and other investigators to be useful in this distinction. The findings that are entirely diagnostic for DA are perineural and vascular invasion; however, they are rarely detectable in biopsy specimens. The most common findings that are highly suggestive of DC and can also be expected in biopsy specimens include random distribution of ductal structures, irregular ductal contours, nuclear enlargement (>3 times the size of a lymphocyte), and pleomorphism, distinct nucleoli, and mitosis. Other, somewhat rarer findings are uninterrupted proliferation of numerous (>50) ducts, intraluminal necrotic cellular debris, hyperchromatic raisinoid nucleoli, the presence of naked ducts in fat without surrounding pancreatic elements or fibrous tissue, and ducts lying adjacent to arterioles. Findings that favor a benign process over an invasive carcinoma are: lobular architecture with clusters of evenly spaced ductal units, uniformly sized ductal elements, smooth ductal contours, ducts surrounded by acini or islets, and intraluminal mucoprotein plugs. Combinations of these criteria should aid in the differential diagnosis of invasive ductal adenocarcinoma from benign/reactive ducts in the pancreas. PMID- 16273948 TI - Guidelines for the evaluation and assessment of the sustainable use of resources and of wastes management at healthcare facilities. AB - This paper presents guidelines that can be used by managers of healthcare facilities to evaluate and assess the quality of resources and waste management at their facilities and enabling the principles of sustainable development to be addressed. The guidelines include the following key aspects which need to be considered when completing an assessment. They are: (a) general management; (b) social issues; (c) health and safety; (d) energy and water use; (e) purchasing and supply; (f) waste management (responsibility, segregation, storage and packaging); (g) waste transport; (h) recycling and re-use; (i) waste treatment; and (j) final disposal. They identify actions required to achieve a higher level of performance which can readily be applied to any healthcare facility, irrespective of the local level of social, economic and environmental development. The guidelines are presented, and the characteristics of facilities associated with sustainable (level 4) and unsustainable (level 0) healthcare resource and wastes management are outlined. They have been used to assess a major London hospital, and this highlighted a number of deficiencies in current practice, including a lack of control over purchasing and supply, and very low rates of segregation of municipal solid waste from hazardous healthcare waste. PMID- 16273949 TI - Micrometeorological measurements of N2O and CH4 emissions from a municipal solid waste landfill. AB - Micrometeorological measurements of methane (CH4) and nitrous oxide (N2O) emissions were made at the decommissioned Park Road Landfill in Grimsby, Ontario, Canada between June and August 2002. The influence of precipitation, air temperature, wind speed and barometric pressure on the temporal variability of landfill biogas emissions was assessed. Gas flux measurements were obtained using a micrometeorological mass balance measurement technique [integrated horizontal flux (IHF)] in conjunction with two tunable diode laser trace gas analyser (TDLTGA) systems. This method allows for continuous, non-intrusive measurements of gas flux at high temporal resolution. Mean fluxes of N2O were negligible over the duration of the study (-0.23 to 0.02 microg m(-2) s(-1)). In contrast, mean emissions of CH4 were much greater (80.4 to 450.8 microg m(-2) s(-1)) and varied both spatially and temporally. Spatial variations in CH4 fluxes were observed between grass kill areas (biogas 'hot spots') and the densely grass-covered areas of the landfill. Temporal variations in CH4 fluxes were also observed, due at least in part to barometric pressure, wind speed and precipitation effects. PMID- 16273950 TI - Nuisance flies and landfill activities: an investigation at a West Midlands landfill site. AB - Nuisance-causing flies were studied in and around a West Midlands, UK, landfill site from mid-January 2003 to mid-January 2004. The most important species was the common housefly, Musca domestica, which made up more than 92% of the total catch on traps in premises and was also frequent on the landfill site. An estimated 2 million common houseflies were imported to the site in waste as eggs, larvae or pupae during the peak month, July. Most did not apparently survive to maturity, only about 20 000 per month emerging from the tipped refuse. Lesser houseflies (Fannia canicularis) were also very commonly imported but they seemed unable to survive the conditions on the tip in their immature stages and few were found in emergence traps. No correlation was found between the distance separating premises from the landfill site and the number of M. domestica trapped in those premises. White sticky traps were effective for monitoring changes in fly populations over the longer term whereas the Scudder grill was more useful for making objective 'snapshots' of fly activity. There is scope to improve both the monitoring of fly activity and the investigation of complaints made by the general public about flies. PMID- 16273951 TI - The influence of operational conditions in sequencing batch reactors on removal of nitrogen and organics from municipal landfill leachate. AB - The removal of nitrogen and organics from municipal landfill leachate in sequencing batch reactors (SBR) was investigated in the present study. The influence of hydraulic retention time (HRT), sludge age, manner of leachate dosage (short filling period of SBR and filling during the reaction period), and operational conditions with and without a mixing phase in the SBR cycle was explored. Four series were performed. In each series, the HRT used in the four SBRs was 12, 6, 3 and 2 days, respectively. Series 1 and 2 were characterized by a short leachate filling period, whereas series 3 and 4 were characterized by filling during the 4 h duration of the reaction in the SBR cycle. In series 1-3 SBR reactors worked with mixing and aeration phases, whereas in series 4 they worked only with an aeration phase. The effectiveness of the removal of organics increased with the extension of the HRT of leachate, particularly under operational conditions with the mixing and aeration phases in the SBR cycle. At 12 days HRT, the SBRs with the mixing and aeration phases in the cycle (series 1 3) showed better results than those with only an aeration phase (series 4). However, at 2 days HRT the operational conditions in SBR reactors with leachate filling over the reaction period (series 3 and 4) were more suitable. The highest efficiency of ammonium removal was obtained in series 1 with a short leachate filling period. In this series, at an HRT of 3-12 days, the ammonium concentration in the effluent did not exceed 1 mg NNH4 L(-1). Nitrogen removal proceeded mainly in the aeration phase as a result of ammonium losses and, to a lesser extent, dissimilative nitrate reduction over the mixing phase. The highest percentage of nitrogen removal as a result of ammonium losses was observed in series with a short filling period and long sludge age (series 1) and also in series without a mixing phase and filling over the aeration phase (series 4), whereas the highest nitrogen consumption for biomass production occurred in series 3 with filling during the reaction period and mixing phase of the cycle. PMID- 16273952 TI - A laboratory study on migration of K+ in a two-layer landfill liner system. AB - Contaminant transport through the clay liner and the underIying secondary leachate drain layer (SLDL) in landfills was studied through a laboratory test, and analysis method on the transport of K+ in a two-layer soil system. The soils used for this study were Ariake clay and the underlying layer, Shirasu soil from the Kyushu region of Japan, representing the clay liner material and SLDL material, respectively. The effective diffusion coefficients (De) of the selected target chemical species, potassium (K+) for the Ariake clay and Shirasu soil were back-calculated using a computer program, and it was found that values of De derived from this study were consistent with those previously published. The hypothesis that the mechanical dispersion process can be negligible has been proved to be reasonable based on both the observation that the predicted values fit the experimental data and the analyses of two dimensionless parameters. Parametric analysis showed the transport of K+ through the soils is controlled by advection-diffusion rather than diffusion only, whereas at low Darcy velocity (i.e. < or = 10(-9) m s(-1)), transport of K+ would be controlled by diffusion. The test results and parametric analysis may be applied in design of landfill liners and SLDLs, particularly in coastal areas. PMID- 16273953 TI - Combined incineration of industrial wastes with in-plant residues in fluidized bed utility boilers--decision relevant factors. AB - In Austria more than 50% of the high-calorific industrial residues and wastes generated are utilized for energy recovery in industrial utility boilers. This study investigated full-scale trials of combined incineration of in-plant residues with various industrial wastes. These trials were carried out in order to learn how the alternatively used fuel influences the incineration process itself as well as the quantity and quality of the various incineration products. The currently used fuel, which consisted of in-plant residues as well as externally acquired waste wood and the refuse-derived fuel (RDF) mixtures used during the full-scale trials are characterized in terms of material composition as well as chemical and physical parameters. An input-output mass balance for the incineration plant (two fluidized bed combustion units, 20 and 30 MW, respectively) has been established, based on the data collected during the full scale incineration trials. Furthermore, pollutant concentrations in the off-gas as well as the solid incineration residue are reported. It is not only the pollutant content but also a variety of other internal as well as external factors that have to be considered if a company is to decide whether or not to thermally utilize specific waste types. Therefore a strengths and weaknesses profile for several types of waste and the specific industrial boiler is also presented. PMID- 16273954 TI - Leachability testing of metallic wastes. AB - The performance of two tests, a batch test and a percolation test for the characterization of waste as suggested in the EU council decision 2003/33/EC was investigated. The tests were carried out on two solid waste streams from a metal recycling industry. The concentrations of heavy metals such as Cu, Zn and Pb were more than one order of magnitude lower than the proposed limit values. Generally, batch test values were equal or higher than percolation test values. With the proposed test procedures both materials could be considered as non-dangerous wastes. The test performance was also investigated using a leachant with higher ionic strength instead of demineralized water as prescribed. The results clearly show a significant increase in the concentration of some heavy metals. Total concentrations of phenolic compounds and polychlorinated biphenyls were less than 1 p.p.m. and 2 p.p.b., respectively. The precision of the batch and the percolation tests were on average 48 and 35%, respectively. PMID- 16273955 TI - Utilization of bottom ash from the incineration of separated wastes as a cement substitute. AB - Waste incineration is still an essential technology in the concept of integrated waste management. Most of the combustion residues are incinerator bottom ash. It has been discovered that incinerator bottom ash from the incineration of separated waste in the primary chamber of the modular two-stage incinerator mainly consists of metal oxides, especially SiO2 and CaO, in proportions that are quite similar to those in cement and so the feasibility of its application as a substitute for cement in concrete was investigated. It was found that after 28 days, the flexural and compressive strengths of the binder using bottom ash were practically comparable with those of a pure cement mixture. The results show that it is reasonable to use a binder containing incinerator bottom ash for applications in which an early-stage lower strength of concrete element is acceptable. PMID- 16273956 TI - Investigation of briquetting of metal waste from the bearing industry. AB - An economical method to process the metal waste that comes from the ball-bearing industry is presented. The purpose of the study was to determine the physical chemical properties of the material, to present the most suitable binders and identify the factors that can affect briquette strength. The mechanical strength and resistance to gravitational drop were defined for both fresh briquettes and those that had been seasoned. The briquette structure was also tested. On the basis of the results of experimental studies and laboratory trials two techniques for processing the waste from the ballbearing industry on an industrial technological scale were developed. The economic and ecological impacts of these industrial applications were examined. The results of the investigations suggest that the briquettes might be recycled in steel-making furnaces. The reported solution to the problem of management of this type of waste appears to be universal and could also be applied by other waste-related enterprises. PMID- 16273957 TI - Estimation and allocation of solid waste to bin through geographical information systems. AB - This study presents a geographical information system (GIS)-based procedure for the precise estimation of solid waste generation, computed using the local population density and income group distribution. Using a triangulated irregular network (TIN) in a GIS environment, the procedure further determines the command area for waste allocation to a particular bin which is generally located so the route slopes towards the collection points for ease of transportation by cart pullers. Computational results of bin location, type, size and the frequency of removal are presented for a typical urban area with known population density, income group distribution, road network and topology. PMID- 16273958 TI - Long workhours, work scheduling and work-related injuries among construction workers in the United States. AB - OBJECTIVES: The objectives of this study were (i) to examine work scheduling in construction and (ii) to establish whether there is any connection between workhours and safety outcomes among construction workers. METHODS: The National Longitudinal Survey of Youth, 1979 cohort (NLSY79), was used for the data analysis. Odds ratios were used to measure the risk of work-related injury in different worker groups. RESULTS: The findings showed that (i) construction workers started work earlier, worked longer days and fewer weeks a year, and were more likely to hold multiple jobs and change jobs than their nonconstruction counterparts and (ii) long workhours and irregular work schedules were significantly associated with a higher work-related injury rate after control for possible confounders. CONCLUSION: The results provide evidence that overtime and irregular work scheduling have an adverse effect on worker safety. PMID- 16273959 TI - Stressful life events and occupational accidents. AB - OBJECTIVE: The purpose of this study was to examine the association between stressful life events and occupational accidents. METHODS: This was a population based case-control study, carried out in the city of Botucatu, in southeast Brazil. The cases consisted of 108 workers who had recently experienced occupational accidents. Each case was matched with three controls. The cases and controls answered a questionnaire about recent exposure to stressful life events. RESULTS: Reporting of "environmental problems", "being a victim of assault", "not having enough food at home" and "nonoccupational fatigue" were found to be risk factors for work-related accidents with estimated incidence rate ratios of 1.4 [95% confidence interval (95% CI) 1.1-1.7], 1.3 (95% CI 1.1-1.7), 1.3 (95% CI 1.1 1.6), and 1.4 (95% CI 1.2-1.7) respectively. CONCLUSIONS: The findings of the study suggested that nonwork variables contribute to occupational accidents, thus broadening the understanding of these phenomena, which can support new approaches to the prevention of occupational accidents. PMID- 16273960 TI - Occupational risk of affective and stress-related disorders in the Danish workforce. AB - OBJECTIVES: A population-based, nested, case-control study was carried out to quantify the risk of affective and stress-related disorders according to occupation in the entire Danish workforce. METHODS: All incident hospital patients and out-patients aged 18-65 years who received a first-time-ever diagnosis of an affective disorder (ICD-10, F 30-39) or stress-related condition (ICD-10, F 40-48) in Denmark from 1 January 1995 through 31 December 1998 were identified in the Danish Psychiatric Central Research Register (N=28 971). For each case, five randomly selected referents of the same age and gender were drawn from a 5% sample of the Danish population (N= 144 855). The occupation held 1 year before a person became a case was obtained from Denmark's Integrated Database for Labour Market Research. Occupation was classified according to the Danish version of the International Standard Classification of Occupations (ICD). Relative risks (RR) and 95% confidence intervals (95% CI) for 25 occupational categories with clerical staff as the reference were calculated using a conditional logistic regression adjusted for sociodemographic covariates. RESULTS: Eight occupations were associated with significantly elevated risks (RR range 1.20-1.58) among the women, while eight occupations were associated with a significantly reduced risk (RR range 0.50-0.76) among the men. The risks were highest for the teaching (RR 1.58) and health (RR 1.53) professions. Only social workers and professionals caring for mentally and physically disabled persons faced an elevated risk irrespective of gender (women RR 1.72, 95% CI 1.38-2.16; men RR 2.09, 95% CI 1.38-3-15). CONCLUSIONS: Major depression and stress-related psychiatric disorders are related to occupation. Risk profiles vary strongly according to gender. PMID- 16273961 TI - Association between work-related psychological stress and arterial stiffness measured by brachial-ankle pulse-wave velocity in young Japanese males from an information service company. AB - OBJECTIVES: This study examined the relationship between work-related psychological stress and arterial stiffness in young Japanese workers. METHODS: A cross-sectional study was conducted on 396 Japanese male workers, aged 24 to 39 years, employed in a Japanese information service company. Work-related psychological stress was measured by the Job Content Questionnaire based on the job demand-control model. The job-strain index was defined as the ratio of job demand to job-control scores. The outcome of the study was the degree of arteriosclerosis as assessed by brachial pulse-wave velocity (baPWV). The cardiovascular risk factors analyzed were age, heart rate, blood pressure, body mass index, serum lipid, blood sugar levels, catecholamine levels, ethanol consumption, smoking, and overtime. In addition, psychological responses were assessed by tension-anxiety and anger-hostility scales in the Profile of Mood States (POMS). RESULTS: The baPWV was positively (P<0.05) associated with physiological variables including age, heart rate, body mass index, and serum levels of total cholesterol, fasting glucose, and noradrenaline, but negatively (P<0.01) associated with the job-strain index. Significant associations were not found on the POMS tension-anxiety and anger-hostility scale scores. The negative correlation between baPWV and the job-strain index was consistent even after control for the effects of significant physiological variables. CONCLUSIONS: The association between job stress and baPWV was found to be inconsistent with the results of previous western studies, and it may require further investigation while taking into account occupation, cardiovascular risk factors, and Japanese culture. PMID- 16273962 TI - Acute effects of cigarette smoking on the heart rate variability of taxi drivers during work. AB - OBJECTIVES: This study attempted to clarify the acute effects of cigarette smoking on autonomic nervous function among taxi drivers under ordinary work conditions. METHODS: Holter electrocardiographic recordings from 20 healthy middle-aged taxi drivers were analyzed for the time from 0800 in the morning to 0159 at night. The amplitudes (milliseconds) of the high-frequency (HF) component and the ratio of the low-frequency component to HF (LF/HF) were calculated as changes in the R-R interval, and time-course changes were investigated by a complex demodulation method. The exact starting time of smoking was identified with the use of a specially designed cigarette lighter. The mean LF/HF and HF for 5 minutes immediately prior to smoking were calculated as the baseline, and the means for every 5 minutes up to 15 minutes were calculated. The average values of these parameters for all of the cigarette smoking within the same time span of 0800-1659 and 1700-0159 were also determined. RESULTS: The LF/HF significantly increased (P<0.05) within 5 minutes from the baseline immediately after smoking. This significant change in LF/HF was observed only at night. Although the interactive effect of the time of day on time course changes was not significant for the LF/HF or HF, the reactivity to increase LF/HF and decrease HF was more prominent at night. CONCLUSIONS: Cigarette smoking significantly increased LF/HF within 5 minutes during ordinary taxi driving. Nighttime smoking seemed to have a more potent acute effect on the cardiac modulation of taxi drivers than in the daytime. The sympathomimetic and parasympatho-withdrawal response of smoking may play an additional role in increasing cardiac risk among taxi drivers. PMID- 16273963 TI - Reduced productivity after sickness absence due to musculoskeletal disorders and its relation to health outcomes. AB - OBJECTIVES: This prospective cohort study quantified the reduced productivity of workers on full duty after sickness absence from a musculoskeletal disorder and determined the effect of health parameters such as perceived pain, functional disability, and general health on reduced productivity. METHODS: Workers were included who were returning to work from 2- to 6-week sickness absence due to a musculoskeletal disorder. Self-administered questionnaires at baseline, after return to work, and at a 12-month follow-up were used to collect information on productivity and health status. Logistic regression analyses evaluated the determinants of reduced productivity and determined the level of productivity loss shortly after return to work. RESULTS: Reduced productivity was prevalent for 60% of the workers after they returned to work, and for 40% still at the 12 month follow-up. The initial musculoskeletal disorder caused 75% of the productivity loss shortly after return to work and 60% at the follow-up. Among those with productivity loss, the median loss for an 8-hour workday was 1.6 hours shortly after return to work and also at the follow-up. Worse physical health, more functional disability, and poorer relations with the supervisor were associated with productivity loss shortly after return to work, whereas recurrent sick leave was the greatest predictor of productivity loss at the follow-up. CONCLUSIONS: Reduced productivity was common among workers returning to full duty after sickness absence due to a musculoskeletal disorder. Productivity loss illustrates the importance of the timing of return to work, especially among workers with residual functional disability after return to work. Moreover, the supervisor should be engaged early in the return-to-work process to guarantee an early, sustainable, and productive return to work for the employee. PMID- 16273964 TI - Job strain and risk of musculoskeletal symptoms among a prospective cohort of occupational computer users. AB - OBJECTIVES: Most previous studies of the association between psychosocial stress and musculoskeletal illness among computer users have been cross-sectional and have yielded inconsistent results. The association between a measure of psychosocial stress, "job strain", and incident neck-shoulder and arm-hand musculoskeletal symptoms was investigated among recently hired computer users. METHODS: The participants worked for one of several large employers and were followed prospectively for 6 months. The "job demands" and "decision latitude" subscales of the Job Content Questionnaire were used to estimate the job-strain quadrants and a ratio measure of job strain which was subsequently categorized. Incident musculoskeletal symptoms were obtained with weekly diaries. Proportional hazards models were used to estimate associations between job strain and incident musculoskeletal symptoms. RESULTS: Those in the high-strain quadrant were at increased risk of neck-shoulder symptoms [hazard ratio (HR) 1.65, 95% confidence interval (95% CI) 0.91-2.99] when compared with those in the low-strain quadrant. Those in the highest strain-ratio category were also at increased risk of neck shoulder symptoms when compared with those in the lowest strain-ratio category (HR 1.52, 95% CI 0.88-2.62). Modification by previous years of computer use was observed, with an elevated risk observed for those in the highest job-strain ratio category who also had low previous computer use (HR 3.16, 95% CI 1.25 8.00). There did not appear to be an association between either measure of job strain and incident arm-hand symptoms. CONCLUSIONS: In this cohort, workers who reported high job strain were more likely to develop neck-shoulder symptoms. PMID- 16273965 TI - Utility of restricted neck movement as a diagnostic criterion in case definition for neck disorders. AB - OBJECTIVES: This study explored the utility of restricted neck movement in epidemiologic case definition for neck disorders. METHODS: Data on neck pain, sensory symptoms in the arm, psychosocial and physical risk factors for neck disorders, and the range of active neck movements were obtained through a self administered questionnaire, interview, and physical examination for a community based sample of 2145 adults aged 25-64 years. The prevalence of neck pain and sensory symptoms was examined according to the extent of neck movements. Logistic regression was used to assess the relation of risk factors to neck disorders, defined by various combinations of pain and restricted movement, and the associations were compared. RESULTS: The ranges of different categories of neck movement were correlated within persons, and their sum ("total neck movement") was unimodally distributed across persons. The prevalence of neck pain and sensory symptoms was elevated when total neck movement was below the 30th centile, and particularly below the 10th centile. Associations between neck pain and low vitality and poor support at work were stronger when there was also restricted neck movement, but the extent of neck movement did not materially modify the relation of neck pain to physical risk factors. CONCLUSIONS: The restriction of total neck movement may serve as a marker of severity in community based studies of neck disorders, but no indication was found that it distinguishes a subset of cases with a distinct etiology. PMID- 16273967 TI - Opportunities and challenges for collaboration in research and practice in injury prevention across work and other settings. AB - Even though injuries at work and in other settings (homes, roads, etc) commonly involve similar mechanisms, research and prevention are usually specific to each setting. The potential benefits of addressing common features of injury prevention are noted across setting, while some unique features of the work environment are acknowledged. Further integration is recommended using the following approaches: (i) organization of research questions by injury mechanisms and (ii) support of comparative research across settings by funding agencies. PMID- 16273966 TI - A new bricklayers' method for use in the construction industry. AB - OBJECTIVES: The aim of this study was to investigate the effect of raised bricklaying on physical workload, reported musculoskeletal disorders, sickness absence, and job satisfaction. METHODS: A controlled intervention study with a follow-up period of 10 months was performed among 202 bricklayers from 25 construction companies. RESULTS: The introduction of devices for raised bricklaying decreased the physical load on the lower back and, to a less extent, on the shoulders and upper extremities. Although raised bricklaying had no effect on the number of lifts, decreases in trunk bending lowered the biomechanical moment. The results showed no decrease in reported musculoskeletal symptoms as a result of the adoption of raised bricklaying. Irrespective of the reason(s), the percentage of bricklayers in the intervention group reporting sickness absence was significantly lower than the same percentage in the control group. The results also showed that, in general, the bricklayers in this study were very satisfied with the use of devices for raised bricklaying. CONCLUSIONS: Controlled intervention studies on ergonomic improvements are rare. This study shows that the introduction of an ergonomic improvement in the construction industry may reduce physical load and the incidence of sickness absence. PMID- 16273968 TI - Toward an integrated medical curriculum. PMID- 16273969 TI - Communication: deaf patients and their physicians. PMID- 16273970 TI - A collaborative management model for mental health care at the Rhode Island Free Clinic. PMID- 16273971 TI - The doctor dilemma: understanding the health workforce in Rhode Island. PMID- 16273972 TI - The health of Rhode Island adults, 2001: results from the Rhode Island Health Interview Survey. PMID- 16273973 TI - Systemic corticosteroids and COPD exacerbations. PMID- 16273974 TI - Judicial diagnosis 'conscience' vs. care how refusal clauses are reshaping the rights revolution. PMID- 16273975 TI - Healthy life expectancy in Rhode Island. PMID- 16273976 TI - The social matrix of adult physical activity in Rhode Island. PMID- 16273977 TI - Working together to improve ICU care in Rhode Island. PMID- 16273978 TI - Capitalism, competition, and commercialism: the healthcare marketplace vs. the medical home. PMID- 16273979 TI - Charting new HMO territory. The action was in the Northeast last week, as WellPoint moved on WellChoice, and HIP-Group Health merger announced. AB - Consolidation in the industry made bigger strides last week, with two large managed-care deals and an information technology merger. While some fear the insurer deals will put even more price pressure on physicians and hospitals, the GE Healthcare-IDX merger drew some praise. James Thrall, left, of Massachusetts General Hospital, says it's likely to improve integration of radiology systems. PMID- 16273980 TI - GE makes big move in IT. Healthcare unit to buy IDX for dollar 1.2 billion. PMID- 16273981 TI - Agenda forges ahead. DeLay scandal not seen as affecting GOP plans. PMID- 16273982 TI - Inching toward EMRs. Fla. Blues, Humana to share patient data over Web. PMID- 16273983 TI - Inundated with patients. Hurricanes boost volume at Houston, La. hospitals. PMID- 16273984 TI - Prices fluctuate wildly: GAO. Study finds no correlation between prices, costs. PMID- 16273985 TI - Outsourcing in and outs. Add concierge services to the tasks being handled by outsiders. For many hospitals, however, outsourcing is still out of bounds. PMID- 16273987 TI - A market doubles--overnight. Baton Rouge hospitals face unprecedented planning and logistical challenges. PMID- 16273988 TI - Good news and bad news. EMR financial gains high,quality gains low: study. PMID- 16273989 TI - Demanding change. Brailer decries healthcare's 'broken,' inefficient system. PMID- 16273990 TI - By the numbers. Largest healthcare financing companies ranked by amount of loans underwritten January through August 2005 (dollar in millions). PMID- 16273991 TI - One hundred years of Alzheimer's disease and the neglected second lesson of Alois Alzheimer on multicausality in dementia. PMID- 16273992 TI - Subjective appraisal of Alzheimer's disease caregiving: the role of self-efficacy and depressive symptoms in the experience of burden. AB - Most studies investigating correlates of Alzheimer's disease (AD) caregiver burden have focused on the role of objective factors as opposed to subjective factors. Although objective variables (e.g., caregiver age, patient dementia severity, functional status) have been shown to be significantly associated with burden, the correlations generally are modest and explain relatively little of the variability in caregiver outcomes. Moreover, many of these objective variables are not modifiable and are of limited use in the development of caregiver interventions. Thus, there continues to be a need to identify powerful and modifiable determinants of caregiver burden. This study examined the role of two subjective factors-self-efficacy and depressive symptoms-as predictors of AD caregiver burden. Based on a sample of 80 AD caregivers, hierarchical regression analyses revealed that self-efficacy and depressive symptoms each had a significant independent effect on the experience of burden even after accounting for objective factors. These findings suggest that caregiver interventions aimed to reduce burden may benefit from the incorporation of specific strategies to increase self-efficacy and decrease depressive symptoms. PMID- 16273993 TI - Perceptions of family caregivers' psychosocial behavior when communicating with spouses who have Alzheimer's disease. AB - The literature for caregivers of persons with Alzheimer's disease (AD) notes the importance of attending to the linguistic and psychosocial dimensions of communication. The objective of this study was to investigate the relationship between psychosocial aspects of caregivers' behavior and communication outcomes for persons with AD and their spousal caregivers. In the first part of the study, we selected 40 segments of audio-recorded conversations between persons with AD and their spouses. We then asked 20 healthy seniors who were not caregivers to listen to and read the conversation segments and evaluate each segment along four psychosocial dimensions. They were also asked to independently rate how smooth the communication was in each segment. We hypothesized that when caregivers' speech is perceived to be respectful, caring, not controlling, and/or it ascribes competence to their spouses, it would be associated with more effective communication. Our results supported these hypotheses in that communication was more likely to be rated higher in smoothness when the content and manner of caregivers' speech were perceived to have positive psychosocial qualities. The findings have implications for training caregivers on effective psychosocial behavior when interacting with persons who have AD. PMID- 16273994 TI - Ask the consumer: an innovative approach to dementia-related adult day service evaluation. AB - Historically, family caregivers have been considered the "consumers" when evaluating respite programs for persons with dementia offered by adult day service (ADS) centers. The purpose of this article is to describe a unique evaluation of ADS conducted directly with persons with dementia. Seventeen persons who regularly attended the Silver Club, an ADS program associated with the University of Michigan Turner Geriatric Clinic, were interviewed by an independent, trained interviewer using a single group, one-time, cross-sectional administration of a consumer satisfaction survey. Fifteen persons were able to complete the interview successfully. The implication of this evaluation is that when specially designed procedures are used, persons with dementia are capable of contributing usable data to consumer satisfaction surveys. Including the voice of this vulnerable population improves the quality of an agency 's overall evaluation process and supports the basic philosophy of ADS to preserve the self worth, independence, and dignity of cognitively impaired individuals. PMID- 16273995 TI - Effects of galantamine on working memory and global functioning in patients with mild cognitive impairment: a double-blind placebo-controlled study. AB - Mild cognitive impairment (MCI) causes memory impairment and executive function deficits in those with the condition. There is also some evidence that MCI patients are impaired in their daily functioning. Cholinesterase inhibitors have been widely used for patients with Alzheimer's disease (AD), with evidence of improving cognitive function. There is currently no established treatment for MCI, and cholinesterase inhibitors are beginning to be studied in these patients. Galantamine is a cholinesterase inhibitor that also has nicotinic receptor modulating properties that has been successful in improving AD patients. This study examined the effects of galantamine in patients with MCI in areas of memory, executive functioning, and global functioning. There was a significant improvement in scores on the Functional Activities Questionnaire, which is a measure of global functioning. There were also improvements in the galantamine group on two of six measures in the Cambridge Automated Neuropsychiatric Test Assessment Battery and in immediate free recall on the California Verbal Learning Test. PMID- 16273997 TI - Alpha-Synuclein aggregation in pathological aging and Alzheimer's disease: the impact of beta-amyloid plaque level. AB - In central nervous system diseases, abnormal aggregation of one protein is often associated with aggregation of other proteins. To begin to assess whether beta amyloid (Abeta) is associated with alpha-synuclein (AS) aggregation [secondary Lewy body (LB) formation], we used immunohistochemical techniques to compare the amygdala of 11 subjects with pathological aging and 18 with Alzheimer's disease. Overall, Abeta-40 plaque level was greater in cases with secondary AS aggregates. Abeta-42 plaque level was not associated with AS aggregation. Abeta-40 plaque levels cannot be ruled out as a factor involved in secondary LB formation. PMID- 16273998 TI - Chest imaging. PMID- 16273996 TI - Deglycosylation of anti-beta amyloid antibodies inhibits microglia activation in BV-2 cellular model. AB - Immunotherapy has become a strategy for treatment of Alzheimer's disease, by inducing antibody response to amyloid-beta peptide (AbetaP) or by passive administration of anti-AbetaP antibodies. Clearance of amyloid plaques involves interaction of immunoglobulin Fc receptor (FcR)-expressing microglia and antibodyopsonized Abeta deposits, stimulating phagocytosis but may promote neuroinflammation. Carbohydrate moiety of Fc of the immunoglobulin G molecule plays a significant role in modulating binding to FcR and its effector functions. Here, we enzymatically removed Fc glycan from monoclonal antibody 196 raised against AbetaP Antigen binding ability and in vitro stability of deglycosylated antibody were unaffected by deglycosylation. Moreover, the deglycosylated antibody exhibits low affinity to FcR on microglial BV-2 cells and has limited ability to mediate microglial chemotaxis and antibodydependent cytotoxicity compared to native antibody. These data suggest that deglycosylation of anti Abeta antibodies before in vivo administration might prevent microglial overactivation, thus reducing the risk of neuroinflammatory response during passive immunization. PMID- 16273999 TI - Computed tomography and pulmonary embolus: a review. AB - Pulmonary embolus (PE) is a common clinical condition with significant morbidity and mortality that remains a diagnostic challenge for clinicians. Although many tests exist, multi-detector computed tomography (MDCT) pulmonary angiography has recently emerged as the test of choice. MDCT technique, diagnostic criteria, prognosis assessment and common causes of misdiagnosis are reviewed with a look toward future trends. PMID- 16274000 TI - CT of nontraumatic thoracic aortic emergencies. AB - Computed tomography (CT), especially multidetector row CT (MDCT), is often the preferred imaging test used for evaluation of nontraumatic thoracic aortic abnormalities. Unenhanced images, usually followed by contrast-enhanced arterial imaging, allow for rapid detailed aortic assessment. Understanding the spectrum of acute thoracic aortic conditions which may present similarly (aortic dissection, aneurysm rupture, penetrating atherosclerotic ulcer, intramural hematoma) will ensure that patients are diagnosed and treated appropriately. Familiarity with imaging protocols and potential mimics will prevent confusion of normal anatomy and variants with aortic disease. PMID- 16274001 TI - Imaging of acute thoracic injury: the advent of MDCT screening. AB - Chest radiography remains the primary screening study for the assessment of victims of chest trauma, but computed tomography (CT), particularly multidetector CT (MDCT), has progressively changed the imaging approach to these patients. MDCT acquires thinner sections with greater speed, allowing higher quality axial images and nonaxial reformations than conventional or single-detector helical CT. The speed of MDCT, both in acquiring data and in reconstructing images, makes the performance of total body surveys in the blunt polytrauma patient practicable. In general, CT has been well documented to offer major advantages over chest radiography in both screening for thoracic injuries and in characterizing such injuries. This capacity has been enhanced by the application of multichannel data acquisition. The greater sensitivity of MDCT has been well demonstrated in diagnosing vascular and diaphragmatic injuries. This article reviews current concepts of diagnostic imaging in acute chest trauma from blunt force and penetrating mechanisms emphasizing the spectrum of diagnostic imaging findings for various injuries, based primarily on radiographic and CT appearances. The advantages of MDCT for selected injuries are emphasized. PMID- 16274002 TI - High-resolution CT in diagnosis of diffuse infiltrative lung disease. AB - The chest radiograph remains the first imaging modality for the approach to diffuse infiltrative lung disease (DILD), but, 23 years after its introduction, high-resolution CT (HRCT) is still considered the best imaging tool for the evaluation of the pulmonary interstitium and to diagnose and assess DILD. The introduction of multidetector computed tomography (MDCT) has provided the thoracic radiologist with a powerful tool with which to image the lung. Moreover MDCT has enabled radiologists to understand better the functional information contained within CT images of DILD. By focusing on the HRCT signs, patterns, and distributions of abnormalities, and mentioning the clinical aspects and the new recent advances in pulmonary imaging, in this article we provide an overview of a practical approach to the interpretation of the DILD. PMID- 16274003 TI - An update of CT screening for lung cancer. AB - Currently available results on CT screening for lung cancer show that 1) the work up on baseline screening can be confined to less than 15% of the individuals and to less than 6% on annual repeat screening, 2) almost all cases are detected by screening with very few diagnoses made between screening on the prompting of symptoms, and 3) over 80% of all the diagnoses are of Stage I. This diagnostic performance results from following the I-ELCAP regimen of screening which defines a positive result of the initial CT in the regimen as well as the work-up leading to a diagnosis of lung cancer. The diagnostic performance raises prognostic questions as to the genuineness and curability of these screen-diagnosed lung cancers. All diagnoses of malignancy were confirmed by an expert pathology review and found to represent genuine lung cancer as defined by the 2004 World Health Organization pathologic criteria. Estimates based on growth rates suggest that about 90% of the baseline-diagnosed Stage I cancers are genuine cancers as are essentially all of those diagnosed on annual repeat screening. Preliminary results of the curability of genuine screen-diagnosed Stage I lung cancers indicate a high curability rate of more than 90%. This suggests that more than a high proportion of deaths from lung cancer can be prevented by CT screening followed by early resection. PMID- 16274005 TI - Virtual bronchoscopy: accuracy and usefulness--an overview. AB - Multidetector CT generated virtual bronchoscopy (VB) represents one of the most recent developments in three-dimensional (3D) visualization techniques which allows a 3D evaluation of the airways down to the sixth- to seventh-generation. In comparison with real bronchoscopy, VB has some advantages: it is a non invasive procedure that can visualize areas inaccessible to the flexible bronchoscope. Virtual bronchoscopy is able to evaluate bronchial stenosis and obstruction caused by both endoluminal pathology (tumor, mucus, foreign bodies) and external compression (anatomical structures, tumor, lymph nodes), can be helpful in the preoperative planning of stent placement and can be used to evaluate surgical sutures after lung transplantations, lobectomy or pneumectomy. In children, in some indications, VB can replace fiber optical bronchoscopy (FB) when this technique is considered too invasive. Finally, VB can also be used to evaluate anatomical malformations and bronchial variants. Virtual bronchoscopy is accurate but its accuracy is not 100% because false-positives and false-negatives occur. Virtual bronchoscopy contributes to a better understanding of tracheo bronchial pathology. Fiber optical bronchoscopy will, without doubt, remain the golden standard but it can be expected that in the near future, the technique of VB will find a place in the daily routine. PMID- 16274004 TI - Computer-aided diagnosis in thoracic CT. AB - Computer-aided diagnosis (CAD) provides a computerized diagnostic result as a "second opinion" to assist radiologists in the diagnosis of various diseases by use of medical images. CAD has become a practical clinical approach in diagnostic radiology, although, at present, primarily in the area of detection of breast cancer in mammograms. Currently, a large research effort has been devoted to the detection and classification of various lung diseases in thoracic computed tomography (CT) images. We describe in this article the current status of the development of CAD schemes in thoracic CT, including nodule detection, distinction between benign and malignant nodules, and detection, characterization, and differential diagnosis of diffuse lung disease. Observer performance studies indicate that these CAD schemes would be useful in clinical practice by providing radiologists with computer output as a "second opinion." PMID- 16274007 TI - Population dynamics of Fasciola gigantica in cattle slaughtered in Uyo, Nigeria. PMID- 16274006 TI - Reduction of Theileria annulata infection in ticks fed on calves immunized with purified larval antigens of Hyalomma anatolicum anatolicum. AB - Larval antigen of Hyalonmma anatolicum anatolicum, the vector of Theileria annulata, was purified by two-step affinity chromatography using anti-tick gut specific rabbit IgG and IgG from immunized cattle. The purified antigen showed the presence of a single polypeptide of 37 kDa (GHLAgP) on SDS-PAGE. Two groups (I and II) of naive crossbred calves (Bos taurus x B. indicus) were immunized with I mg of GHLAgP in three divided doses. Immunized calves of group I were also infected with a sublethal dose of T annulata along with a group of non-immunized calves (group III). Animals in groups I, II, III as well a control group (group IV) were challenged with live nymphs of H. a. anatolicum on the 10th day of immunization. There was a significant reduction in the number of emerging adults of 56.9% +/- 1.67%, in calves of group I (p < 0.01) and 63.09% +/- 1.26% in calves of group II (p < 0.001) compared to the controls. The calves of groups I and II showed antibody responses to tick antigen up to day 70 post immunization. Infection with T. annulata was determined in the salivary glands of adult ticks that developed from the nymphs used for challenge infection. In ticks taken from group I calves, there was a 75.0% +/- 0.00% infection compared with only 85.0 +/- 2.88% infection in ticks taken from calves of group III. Using PCR, a lower infection (83.33% +/- 3.33%) was detected in ticks that developed from calves of group I compared with calves from group III (90.00% +/- 2.88%). The ground-up tick supernatants (GUTS) of the ticks taken from calves of group III yielded higher infection rate and exhibited higher infectivity titre in in vitro infection assay of bovine mononuclear cells than the GUTS of the ticks taken from calves of group I. The results suggest a partial reduction in growth rate of T. annulata in ticks feeding on calves immunized with GHLAgP. PMID- 16274008 TI - Serological prevalence of Toxoplasma gondii in free-range chickens from Costa Rica. PMID- 16274009 TI - Reproductive performance of Fogera heifers treated with prostaglandin F2alpha for synchronization of oestrus. PMID- 16274010 TI - The status of selected minerals in soil, forage and beef cattle tissues in a semi arid region of Zimbabwe. AB - Five districts in the Matabeleland region, an arid western area of Zimbabwe, were investigated for the status of Ca, P, Na, Cu and Zn in soil, forage and cattle during the wet and dry seasons over a period of one year. The cattle came from the natural grazing lands and were not supplemented at the time of sampling. Some deficiencies in soil Zn and P were found in the districts of Lupane and Bulilimamangwe, respectively. Dry season soil Ca, Cu and P concentrations were significantly higher (p < 0.05) than rainy season values owing to leaching in all five districts. Most forage samples had mineral concentrations below the critical levels known to be adequate for animal requirements. Forage levels of Ca, Na. Cu and Zn significantly increased (p < 0.05) with advancing maturity, while P significantly decreased (p < 0.05) in almost all the districts. Marked deficiencies of minerals were found in cattle tissues and these levels followed the seasonal trend seen in the forage. These results indicate that cattle in Matebeleland are deficient in P, Ca, Cu and Zn and that grazing areas in the region cannot provide adequate levels of the five minerals studied. PMID- 16274011 TI - Ovarian follicular dynamics during anoestrus in Anglo-Nubian and Saanen goats raised in tropical climate. PMID- 16274013 TI - Some factors affecting reproductive success in crossbred dairy cows on smallholder farms in coastal north-east Tanzania. AB - A 2-year longitudinal survey was carried out to investigate factors affecting reproduction in crossbred cows on smallholder farms in and around an urban centre. Sixty farms were visited at approximately 2-week intervals and details of reproductive traits and body condition score (BCS) were collected. Fifteen farms were within the town (U), 23 farms were approximately 5 km from town (SU). and 22 farms approximately 10 km from town (PU). Sources of variation in reproductive traits were investigated using a general linear model (GLM) by a stepwise forward selection and backward elimination approach to judge important independent variables. Factors considered for the first step of formulation of the model included location (PU, SU and U), type of insemination, calving season, BCS at calving, at 3 months postpartum and at 6 months postpartum, calving year, herd size category, source of labour (hired and family labour), calf rearing method (bucket and partial suckling) and parity number of the cow. The effects of the independent variables identified were then investigated using a non-parametric survival technique. The number of days to first oestrus was increased on the U site (p = 0.045) and when family labour was used (p = 0.02). The non-parametric test confirmed the effect of site (p = 0.059), but effect of labour was not significant. The number of days from calving to conception was reduced by hiring labour (p = 0.003) and using natural service (p = 0.028). The non-parametric test confirmed the effects of type of insemination (p = 0.0001) while also identifying extended calving intervals on U and SU sites (p = 0.014). Labour source was again non-significant. Calving interval was prolonged on U and SU sites (p = 0.021), by the use of AI (p = 0.031) and by the use of family labour (p = 0.001). The nonparametric test confirmed the effect of site (p = 0.008) and insemination type (p > 0.0001) but not of labour source. It was concluded that under favourable conditions (PU site, hired labour and natural service) calving intervals of around 440 days could be achieved. PMID- 16274012 TI - Some factors affecting variation in milk yield in crossbred dairy cows on smallholder farms in north-east Tanzania. AB - A 2-year longitudinal survey was carried out to investigate factors affecting milk yield in crossbred cows on smallholder farms in and around an urban centre. Sixty farms were visited at approximately 2-week intervals and details of milk yield, body condition score (BCS) and heart girth measurements were collected. Fifteen farms were within the town (U), 23 farms were approximately 5 km from town (SU), and 22 farms approximately 10 km from town (PU). Sources of variation in milk yield were investigated using a general linear model by a stepwise forward selection and backward elimination approach to judge important independent variables. Factors considered for the first step of formulation of the model included location (PU, SU and U), calving season, BCS at calving, at 3 months postpartum and at 6 months postpartum, calving year, herd size category, source of labour (hired and family labour), calf rearing method (bucket and partial suckling) and parity number of the cow. Daily milk yield (including milk sucked by calves) was determined by calving year (p < 0.0001), calf rearing method (p = 0.044) and BCS at calving (p < 0.0001). Only BCS at calving contributed to variation in volume of milk sucked by the calf, lactation length and lactation milk yield. BCS at 3 months after calving was improved on farms where labour was hired (p = 0.041) and BCS change from calving to 6 months was more than twice as likely to be negative on U than SU and PU farms. It was concluded that milk production was predominantly associated with BCS at calving, lactation milk yield increasing quadratically from score 1 to 3. BCS at calving may provide a simple, single indicator of the nutritional status of a cow population. PMID- 16274015 TI - Current perspectives. Therapy with organic nitrates: newer ideas, more controversies. AB - Because of their potent hemodynamic effects, organic nitrates have been introduced in the cardiovascular pharmacopoeia since more than a century in the treatment of coronary artery disease and congestive heart failure. Today, nitroglycerin is the most commonly prescribed generic drug in Italy. While their hemodynamic effects and some of their side effects (e.g. nitrate tolerance, the rebound phenomenon) are at least in part known, little is known regarding other effects of nitrates, for instance their antiaggregant, preconditioning-mimetic, and antiatherosclerotic properties. These effects might have tremendous importance in the treatment of cardiovascular patients. At the same time, the effects of nitrate-derived oxygen free radical species require further investigation. This review provides an update on recent findings in this field. PMID- 16274014 TI - Reproductive disorders of crossbred dairy cows in the central highlands of Ethiopia and their effect on reproductive performance. AB - The study was conducted to estimate the proportion of reproductive disorders and to determine factors affecting reproductive performance of crossbred dairy cows under four different production systems in the central highlands of Ethiopia. The principal postpartum reproductive disorders were retained fetal membranes (14.7%) and uterine infection (15.5%). Anoestrus was the major postpartum reproductive problem in the mixed crop-livestock production system (38.6%) and was significantly associated with this production system. Apart from anoestrus, the occurrence of reproductive disorders was not significantly associated with a production system. Most of the reproductive disorders occurred as a complex rather than as a single abnormality. Two or more abnormal conditions were seen in 11.4% of the cases. Each reproductive trait measured was affected adversely by reproductive disorders. Cows with reproductive disorders in each production system, lactation group and suckling and non-suckling group had longer intervals from calving to first service and to conception (p < 0.001) and required more services per conception (p < 0.001). Pregnancy rate and conception to first service were 84.7% and 51.7%, respectively, for cows without reproductive health problems; and 64.2%, and 15.1%, respectively, for cows with reproductive disorders (p < 0.001). Overall, intervals from calving to first service were shorter (p < 0.05) than in younger cows. Intervals from calving to first service and to conception were longer in suckling than in non-suckling cows (p > 0.05). Cows with a good body condition score (> 3.5) at calving had shorter calving to first service and conception intervals than cows in poor condition (p < 0.001). The results showed that reproductive abnormalities, coupled with poor body condition, are important factors that contributed to reproductive inefficiency. An appropriate reproductive health management, a reliable artificial insemination service and supplementary feeding could be the management options to reduce or alleviate some of the problems. PMID- 16274016 TI - Functional assessment of coronary atherosclerosis in the catheterization laboratory: the key role of fractional flow reserve. AB - Patients with suspected coronary artery disease often undergo coronary angiography without prior non-invasive functional stress testing, mainly because of logistic reasons, shortcomings of the non-invasive tests, and a more widespread confidence in invasive techniques. The availability in the catheterization laboratory of the pressure-derived fractional flow reserve (FFR) has provided the interventional cardiologist with the ideal tool for appropriate decision-making based on the functional significance of the coronary stenosis detected at the angiogram. In fact, FFR allows a more refined and individualized understanding of the true severity of coronary artery disease, and, therefore, a more appropriate selection of the epicardial lesions to be treated, especially in patients with dubious lesions and complex disease. A clinical decision-making based on coronary pressure measurement results in a more effective strategy than placing stents on a "trial and error" basis. This is particularly true in case of drug-eluting stents where an approach based on an indiscriminate multi-stenting will annihilate the benefits of these new stents and be unacceptably expensive. In addition, many angiographically mild stenoses happen to be hemodynamically significant and, therefore, deserve revascularization, especially in the drug eluting stent era. PMID- 16274017 TI - Individuals' cardiovascular risk profile projected by family doctors and individuals' knowledge of cardiovascular risk factors: a challenge for primary prevention. The cardiovascular risk prevention project "Help Your Heart Stay Young". AB - BACKGROUND: Whether the practice of family doctors of assessing individuals' cardiovascular risk profile improves individuals' knowledge of risk factors in primary prevention has not been established. Accordingly, we evaluated patients' knowledge of cardiovascular risk factors and lifestyle in healthy subjects whose family doctors provided individual cardiovascular risk score. METHODS: Subjects who visited their family doctor in a time frame of 3 months, who accepted to fill in a simple questionnaire measuring their knowledge of cardiovascular risk factors and of non-pharmacological interventions able to reduce cardiovascular risk were evaluated. RESULTS: Fifty-one family doctors were involved. The study sample comprised 4239 subjects (mean age 56 +/- 9 years, 62% women) in primary prevention. They were classified by their family doctors, based on the Framingham algorithm, as being at low (< 10%; 45.7% of subjects), medium (10-20%; 38.7% of subjects) or moderate-to-high (> 20%; 15.6% of subjects) cardiovascular risk. The prevalence of obese subjects (40, 48, and 49%, respectively) and of heavy smokers (> or = 20 cigarettes/day; 26, 30, and 34%) increased from the low to the moderate-to-high risk group (both p < 0.05). The proportion of subjects unaware of personal history of arterial hypertension (5, 6, and 9%) and that of subjects who were unaware of history of elevated cholesterol levels (10, 11, and 12%, both p < 0.01) increased with higher cardiovascular risk score. The proportion of subjects self-reporting blood pressure > 135/85 mmHg, but self-reporting being normotensive (30, 50, and 52%), and the proportion of subjects who referred cholesterol levels > 200 mg/dl among those who self-referred not to have elevated cholesterol levels (13, 25, and 31%) increased both with cardiovascular risk category (p < 0.001). The proportion of subjects who were unaware of their personal history of diabetes was similar in the cardiovascular risk groups. The prevalence of low educational level was higher (56, 58, and 62%, p < 0.01) and the level of knowledge of non-pharmacological remedies to cardiovascular risk factors (63, 61, and 59%, p < 0.01) was lower in higher cardiovascular risk score group. Subjects aged < 55 years showed similar lack of knowledge about cardiovascular risk factors and the proportion of heavy smoking was as high as in the group of older participants. CONCLUSIONS: In cardiovascular primary prevention, the projection of higher individuals' risk profile by family doctors was not paralleled by an increase in individual's knowledge of major cardiovascular risk factors and of lifestyle interventions able to reduce the cardiovascular risk. PMID- 16274018 TI - Risk stratification and prognosis of patients with known or suspected coronary artery disease by use of supine bicycle exercise stress echocardiography. AB - BACKGROUND: The aim of this study was to assess the long-term predictive values of supine bicycle exercise stress echocardiography (ESE), and the ESE additional role compared to other traditional clinical and rest echocardiographic variables, in 607 patients with low, intermediate and high pretest risk of cardiac events. METHODS: Clinical status and long-term outcome were assessed for a mean period of 46 months (range 12-60 months). ESE was performed for the diagnosis of suspected coronary artery disease (CAD) in 267 patients (43.9%), and for risk stratification of known CAD in 340 patients (56.1%). At baseline, the mean value of wall motion score index (WMSI) was 1.22 +/- 0.36, and the mean left ventricular ejection fraction was 58.5 +/- 10.9%. RESULTS: ESE was positive for ischemia in 210 patients (34.9%), while ECG was suggestive for ischemia in 157 patients (25.8%). During the test only 97 patients (15.9%) experienced angina. At peak effort, the mean WMSI was 1.38 +/- 0.46. A low workload was achieved by 158 patients (26.1%). During the follow-up period there were 222 events, including 82 hard events (36.9%), 48 deaths (21.6%) and 34 acute non-fatal myocardial infarction (15.3%). At stepwise multivariate model, cigarette smoking (p < 0.01), peak WMSI (p < 0.001), ESE positive for ischemia (p < 0.001) and low workload (p < 0.01) were the only independent predictors of cardiac death, while positive ESE, peak WMSI, angina during the test and hypercholesterolemia were the only independent determinants of hard cardiac events. The cumulative 5-year mean survival rate according to ESE response was 95.9% in patients with negative ESE, and 83.7% in patients with positive ESE (log rank 13.6; p < 0.00001). CONCLUSIONS: ESE yields prognostic information in known or suspected CAD, especially in patients with intermediate pretest risk level. The combined evaluation of clinical variables and other ESE variables, such as peak WMSI and exercise capacity, may further select patients at greatest risk of cardiac death in the overall population. PMID- 16274019 TI - Homecare for patients with heart failure in Italy. AB - BACKGROUND; Heart failure (HF) represents an important health issue in western countries, especially for the elderly, frail population. A number of HF patients must usually be assisted at home. No information is available about the usual care of HF patients in Italy. The aim of this study was to describe the characteristics of HF patients receiving homecare in the Italian general practice. METHODS: A questionnaire was sent to 320 general practitioners (GPs) involved in the Health Search project. Among these, 148 (46.2%) answered and 376 home-ridden HF patients (60.3% women, median age 85 years) were identified. RESULTS: 257 (57%) patients were in NYHA class III or IV. Multiple relevant concomitant diseases occurred in 326 (86.7%) subjects. Only 140 (37.2%) patients were able to take their pills without any help; caregivers, mainly family members, were required 24 hours a day in 78.7% of cases. The length of homecare was > 1 year in 84.5% of cases. CONCLUSIONS: According to our data, thousands of HF patients are usually assisted at home for long periods in Italy. This is a very old group of subjects with heavy co-morbidity and a high need for continuous, prolonged assistance. Studies specifically aimed at the care of HF patients are needed. PMID- 16274020 TI - Reverse ventricular remodeling and improved functional capacity after ventricular resynchronization in advanced heart failure. AB - BACKGROUND: Ventricular resynchronization is a non-pharmacological treatment for advanced heart failure refractory to drug therapy and with intraventricular conduction delay. We describe the time course of echocardiographic and functional recovery after resynchronization in 31 patients (mean age 67 +/- 8 years). METHODS: We evaluated NYHA class, echocardiogram, respiratory function, and cardiopulmonary test before pacemaker implantation (baseline), after 1-3 months (short-term evaluation), and 10-15 months afterwards (long-term evaluation, n = 21 patients). Mortality at 1 year was considered. RESULTS: Both at short and long term, patients improved NYHA class, ventricular function, and ventricular volumes. Already at short-term, we observed an increase in oxygen consumption at peak exercise (12.6 +/- 0.6 vs 10.5 +/- 0.5 ml/kg/min), oxygen consumption at anaerobic threshold (9.8 +/- 0.6 vs 8.3 +/- 0.6 ml/kg/min) and oxygen pulse (8.3 +/- 0.5 vs 7.5 +/- 0.5 ml/beat). Ventilatory efficacy (VE/VCO2 slope) and alveolo capillary diffusion (estimated by the measurement of lung diffusion capacity for carbon monoxide - DLCO) improved only at long-term (VE/VCO2: 40.7 +/- 1.6 vs 45.3 +/- 1.8; DLCO: 70.3 +/- 2.7 vs 59.4 +/- 5.9% of predicted, p = 0.05). The 1-year mortality was 9.7%. CONCLUSIONS: Ventricular resynchronization is linked to a fast and prolonged recovery of NYHA class, echocardiographic variables and stress tolerance. The improvement of indexes known to carry a prognostic value confirms that ventricular resynchronization can positively interfere with the evolution of the disease. PMID- 16274021 TI - Atrial fibrillation recurrence after drug-induced typical atrial flutter ablation. AB - BACKGROUND: Catheter ablation of typical atrial flutter (AFL) occurring in patients who take antiarrhythmic drugs for atrial fibrillation (AF) has been proposed as a curative approach for AF. The aim of this study was to evaluate the efficacy of this technique. METHODS: Forty-six consecutive patients (30 males, 16 females, mean age 67 +/- 9 years) with paroxysmal or persistent AF were submitted to right atrial isthmus ablation: 1) 33 patients (group 1) in whom typical AFL spontaneously occurred during oral treatment with propafenone (n = 19), flecainide (n = 9) or amiodarone (n = 6); 2) 13 patients (group 2) submitted to electrophysiological study while taking oral propafenone (n = 3), flecainide (n = 8) or amiodarone (n = 1), in whom sustained AFL was induced (n = 9) or AF was induced and AFL was obtained by intravenous administration of class IC drugs (n = 4). The same antiarrhythmic drug which induced the conversion of AF into AFL was administered after ablation. RESULTS: During a follow-up of 20 +/- 18 months (range 1-78 months), 23 patients (50%) remained asymptomatic and free from AF recurrences. Fifteen patients (33%) with AF recurrences reported a reduction in arrhythmia-related symptoms. Eight patients (17%) did not show symptomatic improvement. These results did not significantly differ between group 1 and group 2. The duration of follow-up was significantly longer in patients with AF recurrence. Among several clinical, echocardiographic and electrophysiological parameters, only atrial enlargement and the absence of structural heart disease were independently associated with AF recurrence. CONCLUSIONS: In selected patients with AF and drug-induced AFL, right atrial isthmus ablation and prosecution of the drug treatment is a safe and feasible approach, which totally eliminates or reduces symptomatic AF recurrences in one half and one third of patients, respectively. However, the number of AF-free patients tends to decrease over time. PMID- 16274022 TI - Lessons from catheter ablation: how a proarrhythmic effect has become a therapeutic chance. The case of class IC/III drugs in atrial flutter. PMID- 16274023 TI - Transesophageal echocardiographic follow-up of pulmonary veins in patients undergoing ostial radiofrequency catheter ablation for atrial fibrillation. AB - BACKGROUND: Ostial radiofrequency catheter ablation (RFCA) of pulmonary veins (PVs) is a promising invasive approach for the non-pharmacologic treatment of atrial fibrillation, but PV stenosis has been reported as a possible complication of this intervention. The aim of this study was to assess PV anatomy and stenosis (i.e. number and progression) by means of transesophageal echocardiography (TEE) during the follow-up of patients undergoing RFCA. METHODS: Twenty-three consecutive patients with refractory and highly symptomatic atrial fibrillation underwent ostial radiofrequency isolation of arrhythmogenic triggers/foci, localized into the PVs, by an electroanatomic approach (CARTO system) or circular mapping with a multipolar catheter (LASSO) placed under radioscopic guidance. All patients were investigated using TEE and magnetic resonance angiography before radiofrequency application to evaluate PV anatomy. TEE examination was repeated after 2 months of follow-up and, in the presence of a stenosis, 1 year later. RESULTS: TEE allowed to identify 100% of the left and right superior PVs, 96% of right inferior PVs, and 74% of the left inferior PVs. Anatomic variants were detected at TEE in 33% of patients against 37% at magnetic resonance angiography (95% of concordance). After ostial RFCA, TEE disclosed a significant reduction in the mean diameters of the left superior PV (14.1 +/- 3.2 vs 12.0 +/- 2.7 mm, p < 0.01), left inferior PV (11.2 +/- 2.3 vs 9.8 +/- 2.2 mm, p = 0.05) and right superior PV (14.2 +/- 2.6 vs 12.9 +/- 2.7 mm, p < 0.05), and an increase in the mean peak velocities of the left superior PV (69.8 +/- 14.8 vs 91 +/- 42.4 cm/s, p < 0.05) and left inferior PV (59.2 +/- 18.1 vs 79.3 +/- 40.5 cm/s, p < 0.05). From a total of 88 PVs treated, 7 (7.9%) showed a higher significant stenosis in patients treated using the LASSO than the CARTO system (31.3 vs 2.8% respectively, p < 0.01). After 1-year follow-up there was no progression of PV stenosis. CONCLUSIONS: TEE was successful to evaluate PV anatomy and stenosis of patients undergoing ostial RFCA for atrial fibrillation. This complication is not rare and seems to be strictly related to the method of ablation, in particular when circular mapping and disconnection of triggers/foci was carried out by only a circular multipolar catheter without an electroanatomic approach. PMID- 16274024 TI - Successful treatment of electrical storm with oral quinidine in Brugada syndrome. AB - A patient implanted with a cardioverter-defibrillator for symptomatic Brugada syndrome was referred to our hospital 17 months later because of recurrent shocks due to ventricular fibrillation (VF). Isoprenaline was intravenously infused and prevented VF episodes, but VF recurred after every attempt of drug discontinuation. A total of 34 shocks were recorded over 25 days. Subsequently, we treated the patient with oral quinidine and the drug suppressed the electrical storm and prevented VF episodes during a follow-up period of 3 years. This case report, together with few others reported in the literature, suggests a role of oral quinidine in the treatment of electrical storm in Brugada syndrome. PMID- 16274025 TI - Sudden heart failure due to a ruptured posterior Valsalva sinus aneurysm into the right atrium: feasibility of catheter closure using the Amplatzer duct occluder. AB - Valsalva sinus aneurysms are usually congenital and relatively rare and tend to be more frequent in adults. Rupture of these aneurysms can result in sudden death or in an abrupt and rapid progressive heart failure. Surgical repair is the traditional treatment of choice. We report the case of a 48-year-old female with a ruptured posterior non-coronary Valsalva sinus aneurysm, resulting in an anomalous aorto-right atrial fistula. Successful percutaneous catheter closure of the massive left-to-right shunt by using the Amplatzer duct occluder is presented. PMID- 16274026 TI - Shaping a vascular graft for total aortic arch replacement. AB - Aortic arch replacement extended to the ascending and/or descending thoracic aorta with a single vascular graft may cause kinking of the prosthesis. We propose an artifact to obtain a curved prosthesis from a straight one for total aortic arch replacement without the risk of kinking. PMID- 16274027 TI - Non-invasive diagnosis of acute ischemic bowel disease detected as hepatic portal venous gas during intra-aortic balloon percutaneous counterpulsation. AB - The authors report a very unusual case of non-invasive diagnosis of acute ischemic bowel disease detected as hepatic portal venous gas during intra-aortic balloon percutaneous counterpulsation. A 64-year-old man with acute ST-elevation myocardial infarction complicated by cardiogenic shock was treated with percutaneous angioplasty and intra-aortic balloon percutaneous counterpulsation. The post-procedural period was complicated by severe abdominal pain. Abdominal computed tomography revealed hepatic portal venous gas. Multiple kidney and splenic ischemic areas were also identified. Colonoscopy showed signs referring to acute ischemic colitis. Computed tomography detection of hepatic portal venous gas has permitted the non-invasive diagnosis of bowel necrosis. PMID- 16274028 TI - Relapsing brucellosis related to pacemaker infection. AB - Infection is a serious complication following pacemaker implantation. Usually it results from normal flora of the skin or from Enterobacteriaceae, Pseudomonas aeruginosa, Streptococcus spp., Enterococcus spp. We report here a case suggesting that Brucella melitensis is able to persist around pacemaker device being a cause of relapsing brucellosis. PMID- 16274029 TI - Two cases of tako-tsubo cardiomyopathy in Caucasians. AB - Tako-tsubo cardiomyopathy is a recently described disease characterized by chest pain, transient left ventricular dysfunction and specific electrocardiographic changes. The disease takes its name from the typical left apical ballooning observed at left ventriculogram. Tako-tsubo cardiomyopathy was first described by Sato in 1990. Since then sporadic cases were reported by Japanese authors, and only a few European publications are available. We describe 2 cases of patients affected by this syndrome. PMID- 16274030 TI - [Development standardized diagnosis and treatment for pancreatic cancer]. AB - The key points on standardized diagnosis and treatment of pancreatic cancer include early detection, preoperative staging, and reasonable resection area and combined therapy, which is important to improve the prognosis of pancreatic cancer in general. PMID- 16274031 TI - [Surgical management of pancreatic cancer: current practice and future prospect]. AB - Pancreatic cancer still represents a serious medical concern for which no adequate solution has thus far been found. Surgical resection, when possible, remains the primary treatment modality and can result in long-term cure. The value of more radical resection remains open to debate, despite the negative results of some recent randomized trials with standard vs. extended lymphadenectomy. More effective patient selection, more rational resection, and more compositive treatment should be emphasized in management strategies. In the future, appropriately designed randomized trials of standard vs. extended resections may confirm the benefit of extended surgical resections. In addition, well powered trials of adjuvant therapies strategies together with surgical resections may identify more effective combinations, which may improve the survival of patients with pancreatic cancer. PMID- 16274032 TI - [Surgical treatment of pancreatic head carcinomas]. AB - OBJECTIVE: To compare the outcomes of patients undergoing surgical treatment for pancreatic head carcinomas during different time course over 40 years in one hospital. METHODS: Totally 346 cases of pancreatic head carcinoma in one hospital were retrospectively analyzed during the periods of 1958-1976, 1977-1987, 1988 1998, and 1999-2003. RESULTS: During the period of 1958-1976, 79 patients with pancreatic head carcinoma were diagnosed and the rate of pancreaticoduodenectomy (PD) was 20.6% (21/79). During the period of 1977-1987, 60 patients with pancreatic head carcinoma were diagnosed and the PD rate was 26.7% (16/60). During the period of 1988-1998, 109 patients with pancreatic head carcinoma were diagnosed and the resection rate was 20.18% (22/109). During the period of 1999 2003, 98 patients with pancreatic head carcinoma were diagnosed and the resection rate was 22.4% (20/98). The total resection rate of pancreatic head carcinomas was 22.8% (79/346). The complication rate of the operation was 42.3% (9/21) during 1958-1976, 37.5% (6/16) during 1977-1987, 27.3% (6/22) during 1988-1998, and 10.0% (2/20) during 1998-2003. The perioperative mortality was 19.0% (4/21) during 1958-1976 (2 cases of pancreatic fistula and 1 case of liver and renal failure), 12.5% (2/16) during 1977-1987 (1 case of multiple organ failure and 1 case of bleeding), and 4.5% (1/22) during 1988-1998 (1 case of multiple organ failure). One patient with bile duct fistula during 1977-1987 and one patient with pancreatic fistula during 1988-1998 were cured by non-operative treatment. There was no perioperative death during 1999-2003. During the period of 1958 1976, the survival rate of patients undergoing PD was 55.5% by 1 year, 23.1% by 3 year, and 11.0% by 5 year. During 1977-1987, it was 56.3% by 1 year, 25.0% by 3 year, and 12.5% by 5 year. During 1988-1998, it was 59.1% by 1 year, 27.2% by 3 year, and 13.6% by 5 year. CONCLUSIONS: Remarkable improvement have been achieved in perioperative preparation and care after surgical treatment of pancreatic head cancer in the past decades. However, the resection rate and prognosis of PC were still poor, although the accuracy of early diagnosis is increasing, and the complications and perioperative mortality of PD are decreasing. PMID- 16274033 TI - [Outcome of extended retroperitoneal lymphadenectomy in the surgical treatment of adenocarcinoma of the head of the pancreas]. AB - OBJECTIVE: To evaluate the clinical outcome of extended retroperitoneal lymphadenectomy as surgical therapy for adenocarcinoma of the head of the pancreas. METHODS: Twenty patients with adenocarcinoma of the head of the pancreas were treated by standard pancreatoduodenectomy (standard group) between 1994 and 1997, and 46 patients with the same disease underwent extended retroperitoneal lymphadenectomy associated with standard pancreatoduodenectomy (radical group) between 1998 and 2002. Clinical and pathological parameters in both groups were reviewed. The postoperative morbidity, mortality, and survival data were compared. RESULTS: The mean total number of lymph nodes resected was significantly higher in the radical group than in the standard group (P < 0.05). Of the 46 patients in the radical group, 26.09% (12/46) had metastatic adenocarcinoma in the resected retroperitoneal lymph nodes. There was one perioperative death in the standard group and two in the radical group. Postoperative diarrhea and lymphatic leakage were only observed in the radical group. Transfusion requirements and postoperative morbidity did not differ between the two groups. The 1-, 2-, and 3-year survival rates were 63.16%, 31.58%, and 21.05% in the standard group, and 65.91%, 37.71%, and 21.21% in the radical group (P > 0.05). When the subgroups of patients with positive lymph nodes were analyzed, the 1-, 2-, and 3-year survival rates were 41.67%, 16.67%, and 8.33% in the standard group, and 64.52%, 32.26%, and 12.9% in the radical group (P < 0.05). A trend toward a better survival was observed in the first 2 years after operation in the radical group, but with no significant differences 2 years later. CONCLUSION: The addition of an extended lymphadenectomy may improve the early survival without increasing the morbidity, but has no significant effect on long-term survival. PMID- 16274034 TI - [Combined multiple organ resection in 16 patients with adenocarcinoma of the body or tail of the pancreas]. AB - OBJECTIVE: To investigate the feasibility and therapeutic results of multiple organ resection in patients with tumor of the body and tail of pancreas. METHODS: The clinical and pathological data were analysed in 16 consecutive patients with neoplasm of the body and tail of pancreas from 1999 to 2004 retrospectively. RESULTS: Multiple organ resection was performed in 6 cases of primary pancreatic adenocarcinoma of the body and tail (3 cases of pancreatic cancer, 2 cases of malignant glucagonoma, and 1 case of well-differentiated pancreatic stromal sarcoma) and 10 cases of extrapancreatic malignancy (4 cases of gastric cancer, 2 cases of gastric leiomyosarcoma, 1 case of duodenal cancer, and 3 cases of colon cancer of hepatic flexure). Distal pancreatectomy with splenectomy was performed in all cases. In addition, 10 patients received splenic flexure colectomy, 6 patients received distal gastrectomy, 3 patients received left nephrectomy, left colectomy, total gastrectomy, liver lobe resection, left adrenalectomy, and local diaphragma resection, and 2 patients received transverse colectomy, subtotal colectomy, proximal proctectomy, proximal gastrectomy, and duodenectomy. No perioperative death and severe complications were observed. Patients with primary pancreatic cancer or pancreatic stromal sarcoma died within 1 year. Two patients with malignant glucagonoma died 51 and 39 months later. The 3-year survival rate was 70% in 10 patients with extrapancreatic malignancy, among which 2 patients with enteric cancer have survived 37 and 48 months. CONCLUSION: Radical combined multiple organ resection may be performed actively in appropriately selected patients. PMID- 16274035 TI - [Clinical comparison on the classical versus extensive Whipple's resection for adenocarcinoma of head of pancreas]. AB - OBJECTIVE: To evaluate the effect of extensive Whipple's resection to the adenocarcinoma of head of pancreas on the survival, complications, and surgical mortality. METHOD: Ninety three patients who received Whipple's surgery between January 1995 and March 2003 were divided into classical group (n = 51) and extensive group (n = 42). Their short-term outcome and survival rate were compared retrospectively. RESULTS: The postoperative complication rate and mortality in classical group and extensive group were 19.61%/3.92% and 16.67%/2.38%, respectively. And 1- and 2- year survival rates in classical group and extensive group were 58.82%/20.59% and 63.33%/23.33%, respectively. CONCLUSIONS: Postoperative complications and mortality will not increase in extensive Whipple's resection for adenocarcinoma of head of pancreas. However, whether extensive Whipple's resection will improve long-term survival still requires further investigation. PMID- 16274036 TI - [Perioperative nutrition support of the patients with pancreatic head cancer]. AB - OBJECTIVE: To explore the effect of perioperative nutrition support on nutritional condition and complications of the patients with postoperative pancreatic head cancer. METHODS: Thirty four patients received perioperative nutrition support, including enteral nutrition and parenteral nutrition (treatment group). Forty eight patients received routine postoperative parenteral nutrition (control group). According to the operative method, these two groups were further divided into two sub-groups: (1) pancreaticoduodenectomy (PD) subgroup, including 13 cases from treatment group, and 24 cases from control group; (2) palliative operation subgroup, including 21 cases from treatment group, and 24 cases from control group. Body weight, total protein (TP), serum albumin (ALB), and the complications after operation were compared. RESULTS: The concentrations of ALB and TP in the treatment group were significantly higher than those in the control group (P< 0.05). Body weight and TP of the patients received PD in the treatment group were significantly better than those of the control group (P < 0.05). CONCLUSION: Perioperative nutrition support can improve postoperative nutritional condition and reduce the postoperative complications in patients with pancreatic head cancer. PMID- 16274037 TI - [Clinicopathological study of lymphatic vessel density and distribution in pancreatic cancer]. AB - OBJECTIVE: To clarify the clinicopathological significance of lymphatic vessel density (LVD) and distribution in pancreatic cancer. METHODS: We measured LVD in 43 pancreatic cancer specimens by immunostaining with specific lymphatic endothelium marker, and examined their relationship with well-defined clinicopathological variables. RESULTS: Intratumoral LVD (9.4 +/- 10.0) was significantly lower than periturmoral (16.0 +/- 9.7) (P < 0.001) and nontumoral LVD (13.5 +/- 6.0) (P < 0.01). Increased peritumoral LVD correlated significantly with tumor staging (P < 0.05) and lymph node involvement (P < 0.05). CONCLUSION: The lymphatic vessels distribution in pancreatic cancer samples and peritumoral lymphangiogenesis may promote the malignant progression and lymph node metastasis of pancreatic cancer. PMID- 16274038 TI - [Cloning of Human canstatin gene and expression of its recombinant protein]. AB - OBJECTIVE: To clone human canstatin gene and express its recombinant protein. METHODS: The total RNA was extracted from human placenta. The canstatin gene fragment was synthesized and amplified from the total RNA by RT-PCR. The resulting product was cloned into pUCm-T vector and transformed into E.coli DH5alpha through electroporation. The gene was sequenced by the Sanger Dideoxy mediated chain-termination method, and then the canstatin cDNA was cloned into the BamHI and HindIII sites of plasmid pET-22b (+) and transformed into E.coli BL21 where it was induced to express proteins by isopropyl-1-thio-b Dgalactopyranoside (IPTG). RESULTS: The extracted total RNA was separated into three clear bands indicating 28S, 18S, and 5S after electrophoresis. The canstatin gene fragment was synthesized and amplified from the total RNA by RT PCR. The resulting products were cloned into pUCm-T vectors, and then were transformed into E.coli DHSa. After an over night culture, both blue and white colonies were found on the agar plate. Six white colonies were selected and cut by BamHI and HindIII. The plasmids DNA in one white colony showed one band near the location of primary plasmid after digested by BamHI and two bands near the locations of primary plasmid and objective gene fragment after digested by HindIII. The cloned gene in this white colony was sequenced and demonstrated to have the same sequence as that of canstatin gene in GenBank. Then canstatin cDNA was cut down from pUCm-T with BamHI and HindIII and ligated into the vector pET 22b (+). The resultant plasmid pET-22b (+)/canstatin was then transformed into E.coli BL21. White colonies were found on LB agar plate. Seven of them were selected and their plasmids were digested with both BamHI and HindIII. After electrophoresis, all selected colonies showed two specific bands, one was found near the location of primary plasmids, and the other near that of objective gene fragment. After IPTG induction, there was a new protein band about Mr 24 000 on SDS-PAGE. As estimated by densitometry, the percentage of the expressed product over total bacterial proteins was 18.2%, 18.8%, 23.0% and 23.4%, respectively, 1, 2, 3, and 4 hours after induction. CONCLUSION: Human canstatin gene was successfully cloned and its recombinant proteins were expressed in this study. PMID- 16274039 TI - [Establishment, characterization, and biological analysis of pancreatic adenocarcinoma cell strain SW1990/FU]. AB - OBJECTIVE: To establish a 5-fluorouracil (5-FU)-resistant pancreatic adenocarcinoma (PAC) cell strain, and to investigate its biological characteristics. METHODS: The PAC cell strain SW1990 was selected into a multidrug-resistant cell strain stepwise with 5-FU, one of the most common drugs used in PAC chemotherapy, for 12 months and subsequently named SW1990/FU. The cell strain was characterized in terms of morphology, biology, and cross resistance to adriamycin(ADM), mitomycin-C (MMC), and gemcitabine. BHLB/c-m nude mice tumor growth and CEA and CA19-9 levels were analyzed. In addition, karyotyping and FACS analysis were performed in SW1990/FU and SW1990. RESULTS: The SW1990/FU cell strain was 132.7 times more resistant to 5-FU than the parental SW1990 cells, and exhibited cross-resistance to other agents. Compared to the parental cells, SW1990/FU cells exhibited a smaller growth rate, delayed cell-doubling time, and specific changes in chromosomes 18. Tumor diameters in multidrug resistance and parental cells inoculated in in vivo experiments were (1.5 +/- 0.30) cm and (0.8 +/- 0.15) cm, respectively. CONCLUSIONS: Morphological adaptation and intracellular changes can be induced by drug challenge in PAC cells. SW1990/FU may be used as an experimental system for the search to overcome drug resistance and to elucidate possible mechanisms of acquired drug resistance in human pancreatic cancer. PMID- 16274040 TI - [Expression of programmed cell death 4 and its clinicopathological significance in human pancreatic cancer]. AB - OBJECTIVE: To investigate the expression of programmed cell death 4 (PDCD4) protein and its clinicopathological significance in human pancreatic cancer. METHODS: Immunohistochemistry was used to examine the expression of PDCD4 protein in 69 specimens of pancreatic cancer and Western blot in 8 fresh specimens. RESULTS: The expression of PDCD4 protein was significantly lower in all 8 fresh pancreatic cancer tissues than that in non-cancerous tissues detected by Western blot. Compared with non-cancerous pancreatic tissue (> 80% of positive cells), low PDCD4 expression was shown in 69 pancreatic cancer tissues (< 30% of positive cells in 36 cases and 30%-80% of positive expression cells in 33 cases). In the 33 cases with 30% and 80% of positive expression cells, the expression rates of PDCD4 protein were 57.6%, 24.2%, and 18.2% in well, moderately, and poorly differentiated cancers, respectively. In the 36 cases less than 30% of positive expression cells, however, the expression rate of PDCD4 protein in well, moderately, and poorly differentiated cases were 19.4%, 41.7%, and 38.9%, respectively. 67.4% (15/23) of the moderately differentiated cases and 70% (14/20) of the poorly differentiated cases showed < 30% of positive expression cells. Only 26.9% (7/26) of the well differentiated cases, however, showed < 30% of positive expression cells, indicating that low PDCD4 expression was associated with histological grade (P < 0.01). There was no relationship between PDCD4 expression and other clinicopathological parameters including patients' sex, age, and TNM stage. CONCLUSIONS: Expression of PDCD4 protein is low in human pancreatic cancer and is correlated with the differentiation levels of human pancreatic cancer. PDCD4 may play an important role in the occurrence and development of pancreatic carcinomas. PMID- 16274041 TI - [NS398 induced apoptosis in pancreatic carcinoma cell strain BxPC-3 through a COX 2-in dependent pathway]. AB - OBJECTIVE: To investigate the effects of the selective cyclooxygenase-2 (COX-2) inhibitor NS398 on the growth of human pancreatic tumor BxPC-3 cell strain and its possible mechanisms. METHODS: The effect of NS398 on cell growth was assessed by 3- (4,5-dimethylthiazol-2-yl) -2, 5-diphenyl thiazolyl blue (MTT) assay. Apoptosis was determined by fluorescence-activated cell scanning (FACS) analysis and assessment of the floating cell/attached cell ratio. Caspase-3 activation was evaluated by Active Caspase-3 Apoptosis Kit with flow cytometry. Reverse transcriptase-polymerase chain reaction analysis (RT-PCR) and Western blot were used to demonstrate expression levels of COX-1, COX-2 mRNA, and protein, as well as Caspase-3 protein in pancreatic tumor BxPC-3 cell strain. RESULTS: Selective COX-2 inhibitor NS398 significantly decreased cell viability and induced apoptosis in pancreatic tumor BxPC-3 cell strain. The protein expression of Caspase-3 was induced by high-concentration NS398. Caspase-3 activity was strongly activated by NS398. CONCLUSIONS: Selective COX-2 inhibitor NS398 has antiproliferative and proapoptotic potential in pancreatic tumor BxPC-3 cells. Such effect is independent of COX-2, but correlates with Caspase-3 activation. PMID- 16274042 TI - [Drug resistance and activity changes of thioredoxin reductase in pancreatic cancer cells strain SW1990 induced by gemcitabine]. AB - OBJECTIVE: To establish gemcitabine-resistant pancreatic cancer cell strain and study the role of thioredoxin reductase (TrxR) in drug-resistant process. METHODS: Gemcitabine-resistant pancreatic cancer cell strain SW1990/GZ was induced by increasing drug dosage intermittently, then the changes of its biological features and the activity of TrxR were examined. RESULTS: Stable drug resistant SW1990/GZ cell strain was established by culturing with gemcitabine for 9 months. The morphology and growth characteristics of the cell strain changed remarkably. The cells shrunk and became rounder; its endoplasm expanded; granular substances increased; and the doubling-time was prolonged. Resistance of the cell line to gemcitabine, fluorouracil, adriamycin, and mitomycin significantly increased. The TrxR activity of the drug-resistant cells was increased markedly. CONCLUSION: SW1990/GZ has certain multidrug resistance to some chemotherapy drugs, and TrxR plays a role in the drug-resistant process. PMID- 16274043 TI - [Comparison of pBcl-2 and pBax expression in primary invasive ductal pancreatic cancer between Chinese and Japanese patients]. AB - OBJECTIVE: To clarify the clinicopathologic significance of the expression of the Bcl-2 protein (pBcl-2) and the Bax protein (pBax), and their clinical implications in Chinese and Japanese patients with human invasive ductal carcinomas (IDCs) of the pancreas. METHODS: The study included 59 Chinese and 65 Japanese patients with IDCs of the pancreas. pBcl-2 and pBax expression were immuno-stained with streptavidin-biotin (SAB) method. RESULTS: pBcl-2 (+) was seen in 35.6% of Chinese and in 23.1% of Japanese patients. pBax (+) was seen in 49.2% of Chinese and 64.7% of Japanese patients. A comparison between them showed that there were significant differences in the male patients, in the patients with the moderately differentiated cancer, and in the elderly patients (chi squared = 4.447, P = 0.035; chi squared = 4.114, P = 0.043; chi squared = 6.657, P = 0.010 respective). In both Chinese and Japanese patients, those with pBcl-2 positive expression had a significantly higher survival rate than those with negative one (chi squared = 9.99, P = 0.0016; chi squared = 7.63, P = 0.0058). The group with pBax positive expression had a significantly higher survival rate in Japanese patients (chi squared = 9.37, P = 0.0022). Japanese patients whose tumors exhibited pBcl-2 and pBax positive immunostaining survived significantly longer than Chinese patients did (chi squared = 4.48, P = 0.0342; chi squared = 5.23, P = 0.023). CONCLUSIONS: The expressions of both pBcl-2 and pBax are high found in Chinese and Japanese patients. The pBcl-2 positive expression implies a better prognosis in both Chinese and Japanese patients with IDCs of the pancreas. The effect of pBax expression on prognosis is different between Chinese and Japanese patients. PMID- 16274044 TI - [Expression and diagnostic value of mesothelin in specimen of pancreas fine needle aspiration]. AB - OBJECTIVE: To investigate the expression of mesothelin in specimen of pancreas fine-needle aspiration and to evaluate the potential contribution of immunohistochemical labeling of mesothelin to the interpretation of pancreas fine needle aspiration (FNA). METHODS: Specimens from 27 patients were selected for immunolabeling. Immunohistochemical EnVision method was used to detect the expression of mesothelin in specimen of pancreas fine-needle aspiration. The labeling in each patient was scored as positive or negative. These results were compared with the cytologic diagnosis and the follow-up data. RESULTS: Nineteen of the 27 patients were ultimately shown to have an adenocarcinoma, and 8 had no evidence of malignancy on follow-up. Initial cytologic diagnosis of malignancy correlated with carcinoma on follow-up in 10 of 10 cases, and initial benign cytologic diagnosis correlated with benign follow-up in 4 of 6 cases. Seven of the 11 patients with suspicious cytology were found to have carcinomas on follow up. Mesothelin labeling was seen in 14 of the 19 patients ultimately shown to have carcinomas and was absent in 7 of the 8 benign lesions (sensitivity, 73.7%; specificity, 87.5%). Five of the 7 cytologically suspicious cases with malignant follow-up labeled for mesothelin. Positive mesothelin labeling was observed in one of the 4 suspicious cases who finally proved to be benign during follow-up. CONCLUSION: Immunohistochemical labeling for mesothelin may be a highly specific tool for the detection of pancreatic adenocarcinoma in FNA specimens and is useful in categorizing cytologically suspicious lesions. PMID- 16274045 TI - [Metallothionein and its isoform genes expression in the human pancreatic cancer cell strains and their function]. AB - OBJECTIVE: To compare the expression of metallothionein (MT) genes and proteins in six human pancreatic cancer cell strains and two human pancreatic cancer drug resistant cell strains and to explore the relationship between the expression of the MT and pancreatic cancer cell chemo-resistance. METHODS: Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to determine the MT isoform-specific mRNA, and cadmium/hemoglobin saturation-electrochemistry to determine MT protein levels. RESULTS: MT protein expression in the pancreatic cancer cell strains was encoded by MT-1A, MT-1B, MT-1E, MT-1F, MT-1G, MT-1X, and MT-2A genes. The expression of MT proteins was upregulated and MT-1B, MT-1E, MT 1X, MT-2A genes overexpressed in human pancreatic cancer drug-resistant cell lines (P < 0.05). CONCLUSION: Expressions of MT proteins and genes correlate with the proliferation and chemoresistance of human pancreatic cancer cell strains. PMID- 16274046 TI - [Clinical significance of the expression of MUC4 mRNA in peripheral blood mononuclear cells of pancreatic cancer patients]. AB - OBJECTIVE: To explore the expression and clinical significance of MUC4 mRNA in peripheral blood mononuclear cells of pancreatic cancer patients. METHODS: The expression of MUC4 mRNA in peripheral blood mononuclear cells of pancreatic cancer patients were detected with reverse transcription realtime PCR. RESULTS: Expression of MUC4 mRNA was not detected in the peripheral blood mononuclear cells of chronic pancreatitis patients and normal healthy people, but was observed in those of pancreatic cancer patients. The positive expression rate of MUC4 mRNA in pancreatic cancer patients was 60%, which was significantly higher than those of chronic pancreatitis patients and normal healthy people (P < 0.01). The positive expression rate of MUC4 mRNA increased with the development of clinical stage, and the positive expression rate in stage of II - IV (TNM system) was 76.92%, which was significantly higher than that of I - II stage (P < 0.01). CONCLUSIONS: Expression of MUC4 mRNA is highly correlated with the clinical stage in pancreatic cancer patients. Detecting the expression of MUC4 mRNA in peripheral blood mononuclear cells of pancreatic cancer patients may be helpful for the early diagnosis and differential diagnosis. PMID- 16274047 TI - [Expressions of p53 and Gadd45a proteins in human pancreatic cancer and their clinicopathological significance]. AB - OBJECTIVE: To study the expressions of p53 and Gadd45a proteins and their clinicopathological significance in human pancreatic cancer. METHODS: The expression of p53 and Gadd45a proteins was detected with immunohistochemistry in a series of 59 pancreatic cancers. Their relationships with the clinicopathological parameters including gender, tumor site, TNM stage, histological differentiation, and the prognosis of pancreatic cancer patients were analyzed. RESULTS: The positive expression rate of p53 protein was 67.8% (40/59) and that of Gadd45a protein was 42.4% (25/59). The positive expression rate of p53 protein is significantly higher in patients < 65 years than in patients > or = 65 years (chi squared = 4.711, P = 0.030). Gadd45a expression was not correlated to the age of the patients. No significant difference was found between the expression of p53 proteins and histological differentiation and TNM stage of the tumors. Gadd45a expression was correlated with histological differentiation of pancreatic cancer (chi squared = 10.052, P = 0.007), but not with TNM stage of the tumors. No significant differences in the prognosis were found between the groups with and without p53 expression (chi squared = 0.09, P = 0.764) and the groups with and without Gadd45a expression (chi squared = 0.14, P = 0.704). CONCLUSIONS: Both p53 and Gadd45a are highly expressed in human pancreatic cancer and may be associated with biological features of pancreatic cancer. Their expression alone or co-expression may be not helpful to evaluate the prognosis of patients with pancreatic cancer. PMID- 16274048 TI - [Effects of Ras antisense oligoribonucleotide on multidrug resistance of pancreatic carcinoma Pc-2 cells]. AB - OBJECTIVE: To investigate the role of Ras antisense oligoribonucleotide (ASODN) in multidrug resistance (MDR) of pancreatic carcinoma Pc-2 cells. METHODS: Ras and P-gp expression was suppressed by Ras ASODN. Sensitivity of Pc-2 cells to chemotherapy was determined by the MTT assay. MDR-1 mRNA level was detected by fluorogenic probe quantitative reverse transcription polymerase chain (RT-PCR) method. Flow cytometry (FCM) was used to detect the accumulative concentration of adriamycin (ADR) in the cells. RESULTS: Ras ASODN significantly inhibited the Ras and P-gp expression (P < 0.05), increased the sensitivity of Pc-2 cells to chemotherapeutic agents (P < 0.05), decreased MDR-1 gene level in Pc-2 cells (P < 0.05), and increased the intracellular intake of ADR in Pc-2 cells (P < 0.05). CONCLUSION: Ras ASODN may enhance the sensitivity of multidrug-resistant pancreatic cancer Pc-2 cells to chemotherapeutic agents by regulating MDR-1 gene level. PMID- 16274049 TI - [Observation of the short term therapeutic effectiveness of body gamma-knife in 48 patients with advanced pancreatic carcinoma]. AB - OBJECTIVE: To investigate the short-term therapeutic effectiveness of body gamma knife in patients with advanced pancreatic carcinoma. METHODS: Forty-eight patients with advanced pancreatic carcinoma were treated by body gamma-knife therapy. The dosage distribution and other radiotherapeutic plans were established on the basis of the carcinoma position, clinical target volume, and patient health condition. The isodose curve was 50%-60% and covered about 95% of the target volume. The single dose was 350-450cGy. The radiation was performed once every one or two days for 10-12 times. RESULTS: There were 33 patients with back pain. 63.6% of the patients got completely controlled, 30.3% pain remitted, and 6.1% ineffective after 2 to 18 months of therapy. The analgesic effective rate was 93.9%. Among 28 patients with obstructive jaundice, 21 patients (75.0%) recovered. Among 42 patients who received CT, tumor disappeared in 5 patients (11.9%), tumor size decreased in 30 patients (71.4%), remained unchanged in 5 patients (11.9%), and enlarged in 2 patients (4.8%). The 6-month, 12-month, and 18-month overall survival rates were 77.1%, 37.5%, and 10.4%, respectively. The whole process was tolerable for all patients and no severe side-effect was observed. CONCLUSIONS: Body gamma-knife can achieve a high local control rate and survival rate. Its short-term therapeutic effectiveness is satisfactory. Body gamma-knife is a safe and reliable treatment option for patients with locally advanced pancreatic carcinoma. PMID- 16274051 TI - [Neoadjuvant therapy in pancreatic cancer]. AB - Pancreatic cancer remains a major troublesome clinical problem, with conventional cancer treatments having little impact on disease course. The extent of disease is often classified as localized, locally advanced, and metastatic. Radical operation is the most effective method, but only 15%-20% of patients have resectable disease, and around 20% of them survive to 5 years. For locally advanced, unresectable, and metastatic diseases, palliative treatment is more appropriate, but the median survival in these patients is less than 6 months and the 5-year survival rates are even lower than 4%. Neoadjuvant therapy has been gradually accepted in breast cancer and gastroenterological cancer, and its value in pancreatic cancer has attracted increasing interests. This paper reviews recent advances of neoadjuvant therapy in pancreatic cancer. PMID- 16274050 TI - [Research development of proteomics in pancreatic cancer]. AB - This review covers the development of proteomics in pancreatic cancer, including the research strategy, technology, content, and problems. PMID- 16274052 TI - [Update in the research of gene therapy for pancreatic carcinoma]. AB - With the advances in immunology and molecular biology, new recognition in the pathogenesis, progression, and metastasis of carcinoma have been achieved. Studies on gene therapy for pancreatic carcinoma have been attempted in different ways, such as inhibiting oncogene, activating tumor suppressor gene, inducing apoptosis, applying gene directed enzyme prodrug therapy, and immune activation. New specific target genes and further development of gene technology may bring the break-through in this field. PMID- 16274053 TI - [Photodynamic therapy: perspectives of its use in ophthalmology]. PMID- 16274054 TI - [The first results of phase IIA of clinical studies of photodynamic therapy for subretinal neovascular membranes with photosense]. AB - The subretinal neovascular membrane (SNM) is a common complication of diseases, such as senile macular degeneration, myopia, pseudohistoplasmic syndrome, and many others. There is no algorithm of treatment for this pathology in today's ophthalmological care; however, many authors have arrived at the conclusion that photodynamic therapy (PDT) plays a major role in solving this difficult problem. Fifteen (18 eyes) patients, including 8 (8 eyes) patients diagnosed as having senile macular degeneration and SNM and 7 (10 eyes) patients diagnosed as having complicated high-grade myopia, and 3 were diagnosed as having bilateral SNM. The Russian agent Photosense, 0.05 mg/kg, was used as a photosensitizer. PDT was performed on day 3 after administration of the agent according to the peak of accumulation and the maximum laser concentration gradient at a wavelength of 675 nm and a light dose of 120 J/cm2. Good functional and angiographic results may be observed after the first 3 months of a prospective controlled open-labeled study of the efficiency of PDT with Photosense; however, longer follow-ups and larger groups of patients are required to make a final assessment of this treatment. PMID- 16274056 TI - [Experimental assessment of the capacities of use of photosense. Communication 1. Fluorescence diagnosis of epibulbar and choroid tumors]. AB - Whether Photosense (PS) may be used in the fluorescence diagnosis (FD) of experimental epibulbar and choroid tumors is discussed. The fluorescent ability of Photosense in FD permits detection of advanced malignancies. The kinetics of concentration of PS and the selectivity of its accumulation in malignant versus normal tissues were investigated. PS is shown to be an effective photosensitizer in the FD and, possibly, photodynamic therapy of tumors. PMID- 16274057 TI - [Experimental assessment of the capacities of use of photosense. Communication 2. Photodynamic therapy for epibulbar and choroid tumors]. AB - The study was undertaken to evaluate the efficiency of photodynamic therapy of pigmented choroid and epibulbar melanoma, by using the agent Photosense. Pigmented choroid and epibulbar melanoma were identified in 50 rabbit eyes. After intravenous injection of Photosense, 0.7 mg/kg, the tumors were irradiated at 675 nm with an argon-pumped dye laser at 150 J/cm2. Photodynamic therapy with Photosense may be effective in treating pigmented choroid and epibulbar melanomas. PMID- 16274058 TI - [Experimental (in vivo) study of the capacities of use of radachlorine (chlorine E-6) for photodynamic therapy in ophthalmology]. AB - To study the capacities of ophthalmological application of the photosensitizer Radachlorine (0.35% solution for intravenous injection), the authors carried out a series of in vivo experiments on rabbits. They studied the time parameters of distribution of the agent in the chorioretinal complex and the reaction of newly formed vessels to photodynamic therapy. Radiation of a diode laser at a wavelength of 0.662 microm was used to stimulate the photosensitizer. A video micro-camera adapted to the ocular of a split lamp was applied to fluorescence fixation. Superficial corneal vascularization was used as a model of the neovascular membrane in response to the application of a non-penetrating silk suture. The findings suggest that photodynamic therapy using Radachlorine is safe and effective in treating ocular tissue neovascularization. PMID- 16274055 TI - [Russian photosensitizer radachlorine in photodynamic therapy for epibulbar and choroid tumors: experimental studies]. AB - The authors used the Russian agent Radachlorine as a photosensitizer (PS) in photodynamic therapy (PDT) for epibulbar and chorioid tumors. Analyzing the results suggests that the efficacy of Radachlorine used as PS in a correctly adjusted dose in PDT may be high. The optimization of effects in the experiment was observed when Radachlorine was used in a dose of 3 mg/kg with a light radiation dose of 500 mW/cm2 upon 5-min exposure. However, a rapid relapse in some cases indicates the instability of the effect and makes it necessary to make repeated PS injections and PDT sessions. PMID- 16274059 TI - [Development of a new technology of photodynamic destruction of newly formed corneal vessels via local administration of photosensitizers]. AB - The purpose of the study was to perfect the procedure, based on the local administration of photosensitizers (PS) into the lumen of vascular trunks, for photodynamic destruction of newly formed corneal vessels. It was performed on 24 rabbits in which corneal intergrowth of newly formed vessels was induced, by applying nine No. 8.0 silk sutures. The vasculature formed 3 weeks later was represented as 20-35 large vascular trunks. A special microneedle, 200 microm in diameter, was designed for local administration of PS. Surgery was performed, by applying microsurgical techniques under an "Opton" operating microscope. The study showed the basic possibility of inserting the microneedle into the lumen of vascular trunks at the limbus with the subsequent filling of vessels with PS. It was sufficient for photodynamic effects to irradiate the cornea with light from the illumination lamp of an operating microscope throughout the operation (3-5 min). The most effective method for locally using PS involved its successive administration into 7-8 largest vascular trunks (the whole network was filled in by anastomoses in each case). The administration of PS into one large nutrient vessel was less effective due to the rapid flowing-out of PS from the vascular minor branches. Deuteroporphyrin sodium (dimegin) has a high photodynamic effect. The morphological basis of the effect is total endothelial destruction of newly formed vessels. The performed studies have demonstrated that the local administration of PS is as effective as the conventional technology of photodynamic therapy. The procedure is more adaptable to the problems of eye surgery due to the absence of systemic action on the patient's body. PMID- 16274061 TI - [Optical coherent tomography in the diagnosis of amblyopia]. AB - The central retinal area was studied in children with different types of amblyopia, by using optical coherent tomography. The results of the latter performed in 113 (208 eyes) children with different types of amblyopia were analyzed. Amblyopia was found to be characterized by structural changes in the central retinal area, which were more pronounced in its dysbinocular form. In obscurant amblyopia, these changes depended on the remoteness of deprivation. The preliminary results given suggest that the similar studies are promising in choosing an algorithm of treatment for such a severe abnormality as amblyopia. PMID- 16274060 TI - [A device for fluorescence diagnosis and photodynamic therapy of eye diseases, by using photosense]. AB - By having a high photodynamic effectiveness and an ability of fluorescence, a Photosense photosensibilizer provides a way of combining photodynamic therapy (PDT) and monitoring its control within a session, which enhances the efficiency of treatment for the subretinal neovascular membrane. A slit lamp-based apparatus complex has been developed to employ the methods of fluorescence diagnosis (FD) and PDT, by applying this photosensitizer. The complex comprises an optical adapter that focusing laser radiation on the fundus of the eye in a range of 100 1000 microm, a video adapter that includes color and high-sensitive monochromic video cameras, as well as a personal computer and software that processes video information from the high-sensitive camera and displays the obtained images in real time. The original system of filters provides an image of the eye fundus in the fluorescent and usual color light at once during a FR procedure. The spatial resolution of the developed apparatus was tested on the test object specially devised for these purposes, which was 10 microm. The sensitivity of the complex is sufficient to record slightly fluorescent objects on the fundus of the eye. PMID- 16274062 TI - [A role of laser retinal tomography in the study of patients with congestive optical disks]. AB - A Heidelberg retina tomograph II was used to examine 35 (55 eyes) patients with intracranial hypertension caused by brain tumors. Ophthalmoscopy revealed no changes in the optic disk in 6 eyes, mild, moderate, and severe congestive optic disks (COD) in 6, 14, and 29 eyes, respectively. The volume of the neuroretinal rim (V) and the area of the Optic disk (S) were measured. There was a significant difference in V (p < 0.01) and S (p < or = 0.02) with the intact optic disk, in mild, moderate, and severe COD. One-week-to-one-month follow-ups after tumor removal showed that in 12 patients the positive or negative changes, as evidenced by the readings of a retina tomograph, corresponded to those in the ophthalmoscopic pattern. Laser retinal tomography is an objective and accurate study of the stereometric parameters of COD in patients with intracranial hypertension, which permits a follow-up of COD. PMID- 16274063 TI - [Retinal vascular lesions in systemic lupus erythematosus and secondary antiphospholipid syndrome]. AB - The study was undertaken to determine a possible association of retinal vascular lesion in systemic lupus erythematosus (SLE) with the antiphospholipid syndrome (APS). A hundred and ninety-four patients (160 females and 34 males, whose mean age was 30.7+/-8.9 years) with verified SLE were examined. Group I comprised 67 patients with retinal vascular lesions (Subgroup la (n = 13) with retinal vascular occlusions and Subgroup 1b (n = 54) without occlusions). Group 2 included 127 patients without retinal vascular lesions. APS was detected in 86 patients. Retinal vascular occlusions more frequently occurred in patients with APS (13.9%) than in those without APS (0.9%) (p = 0.0009) and more frequently in patients with APS and thrombocytopenia (24.3%) than in those with APS without thrombocytopenia (6.1%) (p = 0.0359). Extraocular thromboses were more frequently encountered in Subgroup 1a (69.2%) than in Subgroup 1b (33.3%) (p = 0.0399) and in Group 2 (22.8%), (p = 0.0010). Cerebral circulatory disorder (CCD) was observed in 30.7% in Subgroup la, in 14.8% in Subgroup 1b, and in 7.9% in Group 2 (p = 0.0268). Transient ischemic attack (TIA) occurred in 46.2% in Subgroup la, in 24.1% in Subgroup 2a, and in 14.9% in Group 2 (p = 0.0129). Thrombocytopenia was identified in 69.2% in Subgroup 1a, in 22.2% in Subgroup 2a (p = 0.0021), and in 12.6% in Group 2 (p < 0.0000). The frequency of elevated IgG of anticadiolipin (aCL) in Subgroup 1a (80%) exceeded that in Subgroup 1b (37.9%) (p = 0.0309) and Group 2 (30.7%) (p = 0.0096). Another isotype of aCL (IgM 80%) was observed in patients with retinal vascular occlusions, but the differences in this index were insignificant in the groups and subgroups. The association of pathological changes in retinal vessels in the presence of lupus anticoagulant (LA) was particularly noticeable (91.7 and 52% in Subgroup la and Subgroup 1b, respectively (p = 0.0191) and 47.1% in Group 2 (p = 0.0088). There was an association of amaurosis fugax with eyeground occlusions (p < 0.004), CCD and TIA (p < 0.0002), APS (p < 0.0003), and essential hypertension (p < 0.05). Thus, occlusive lesions in the fundus of the eye are a common manifestation of thrombogenesis in SLE in the presence of APS. The frequency of concomitance of retinal vascular occlusions with cerebral circulatory disorders should be referred to as severe manifestations of SLE. Amaurosis fugas is a manifestation of retinal vascular lesion and associated with the presence of APS and elevated blood pressure in patients with SLE. There is an association of an occlusive process in the fundus of the eye with different symptoms of APS and primarily with IgG of aCL, LA, and thrombocytopenia. PMID- 16274064 TI - [Preliminary results of primary implantation of flexible intraocular lenses in infants under 1 year of age]. AB - The paper presents the results of surgical treatment in 12 (12 eyes) infants aged 3 to 11 months who had different forms of unilateral congenital cataract. The surgical techniques involved the following stages: formation of a sclerocorneal tunnel cut, 3.5 mm in width, anterior continuous circular capsulotomy (anterior capsular rhexis), aspiration-irrigation techniques for removal lens masses, implantation of a flexible intraocular lens (IOL) into the capsular sac, graded opening of the posterior capsule with anterior vitrectomy in some infants. The authors could achieve symmetrical capsular rhexis in 75% and intracapsular IOL fixation in all the cases. During 3-month-to-2-year follow-ups after surgery, IOL held its position in the capsular sac; IOL dislocation and lenticular body capture were not observed in any case. PMID- 16274065 TI - [Sympathetic ophthalmia. Clinical and morphological study in 6 cases after penetrating injury and subsequent repeated vitreoretinal operations]. AB - The paper outlines the clinical and morphological picture of 6 sympathizing eyes enucleated in 1996 to 2003 after vitrectomy and other endovitreal interventions into early injured or operated eyes. Resurgery was attempted early after wounds or a primary operation on the average following 20 days. Sympathetic ophthalmia (SO) generally occurred in the posterior uveal tract as panuveitis or posterior uveitis and diagnosed in the late periods. Specific granulomatous inflammation in the uveal tract was observed in all eyes and the morphological feature of sympathizing eyes was the spread of an inflammatory process to the retina with the development of the latter's epithelioid-cell granulomas and adhesive chorioretinitis. The authors also discuss the possibility of additional antigenic stimulation accompanied by a progressive autoimmune process in the eye and by the generalization of a granulomatous inflammation, as well as the role of retinal minor lesions in this process as a source of additional autoantigens during repeated vitreoretinal operations. The latter, as the authors believe, are a risk factor of SO. In this connection, when repeated vutreoretinal interventions should be made in the presence of an uncompleted wound process in the eye and when they should be conducted under the conditions of active immunosuppressive therapy gain in great importance. PMID- 16274066 TI - [Experimental study of relationships of carbotexim-M to the adjacent tissues during its subcutaneous implantation]. AB - A Russian Carbotexim-M carbonic felt was implanted into the subcutaneous pocket on the back of albino nice. Gross and morphological analysis of changes in rejection was first performed during 4 months and the status after material yield and skin defect healing was also analyzed after 4 months. The risk factors of rejection were the excessive volume of an implant, epithelial thickness and metabolism, the abrasive effect of Carbotexim-M particles. These data indicate the potential problems associated with the superficial implantation of the material in clinical practice. PMID- 16274067 TI - [Analysis of the clinical effectiveness of vitrum vision in the tired eye syndrome and senile macular degeneration]. AB - The paper evaluates the clinical effectiveness of Vitrum Vision ("Unifarm") that contains a complex of carotenoids, vitamins, and trace elements, which is adapted to ocular tissue metabolic features, in the tired eye syndrome and senile macular degeneration. The use of Vitrum Vision in patients with the tired eye syndrome improved the basic parameters of photosensitivity and increased the reserve of relative accommodation and in those with myopia enhanced uncorrectable visual acuity, ameliorated the magnitude of subjective visual and ocular symptoms, and increased the duration of a visual load until sensations of eye tiredness appeared. The administration of the drug in all clinical forms of senile macular degeneration increased visual acuity, improved the basic parameters of photosensitivity, enhanced the functional activity of neurons in the central portions of the retina (as shown by electrophysiological studies) while that in the subretinal neovascular membrane diminished retinal edema, improved the patients' general condition, far vision, and increased comfort on reading. PMID- 16274068 TI - [The specific features of contact correction in children with aphakia]. PMID- 16274069 TI - [A procedure for layer therapeutic-and-tectonic keratoplasty in corneal thinning]. PMID- 16274070 TI - [Causes of low visual functions and rehabilitative methods in patients after successful surgery for retinal detachment]. PMID- 16274071 TI - [Pyotr Fyodorovich Brosse]. PMID- 16274073 TI - [Identification of developmental stages of wisdom teeth on orthopantomograms of adolescents and young adults as an aid for forensic-odontological age estimations: predictive values for the chronological age of 18 years]. AB - Dental findings are an important source of information regarding forensic odontologic determination of identity and age. A main part of dental age assessment is the determination of the wisdom teeth' development. The aim of this study was to answer the question whether the development of wisdom teeth can give information for the task of individual age estimation. Thereby, the investigation could contribute to age estimation efforts that are carried out on persons with an unknown age and where it is of high importance to know as exactly as possible the age of a person, e.g. the age of 18 years or older, for determining the punishment for a criminal offence. MATERIAL AND METHODS: The orthopantomograms of 1053 outpatients were evaluated. The patients were aged 14 to 24 years. The developmental stages of the wisdom teeth were determined according to Demirjian. The data were calculated using statistical tools, like descriptive and inferential statistics in order to provide predictive values for the age to be above or below 18 years. RESULTS: The correlation between the developmental stages of wisdom teeth and the chronological age was high in general. Some values allowed the prediction of an age above 18 years with a probability of more than 70% to 80%, in some instances even more than 90%. The most appropriate developmental stage to determine the age of 18 years was A 1/2 (completed root development, apex open). The positive predictive value of this finding was 94%. DISCUSSION: This study provides for the first time predictive values for the estimation whether or not a person is 18 years of age, based on the evaluation of developmental stages of wisdom teeth on orthopantomograms only. These results should be compared to data from other populations gained by the same method. PMID- 16274072 TI - [An unusual case of a double casualty during sexual intercourse]. AB - Sudden death during sexual activity is generally caused by natural diseases, like coronary sclerosis and myocardial infarction. Only a few case reports describe outer circumstances as responsible for the fatal outcome. This is a case report of a couple that drowned within a car sinking into a river. The extraordinary scenery led to the supposition that sexual intercourse was involved with these fatalities. The victims--a young woman and a young man, both had children, the man was married--kept up an extramarital liaison. Both were reported missing by their relatives days before the corpses could be recovered. PMID- 16274074 TI - [Quantification of human DNA from forensic samples via real time PCR by the help of a telomerase assay]. AB - Quantitative data concerning the content of human DNA and the effect of PCR inhibitors in a sample would provide very important information within a forensic DNA analysis. With the help of real-time PCR it is possible to test DNA samples for these influencing factors. However, the amplified DNA segments detected by means of usual TaqMan DNA probes are longer than most of the short tandem repeats to be investigated. Because of possible DNA degradation, a DNA probe located in the human telomerase transcriptase gene was successfully tested on forensic samples in the present study. Its amplified DNA segment is only 94 bases in length and, thus, shorter than the short tandem repeats. Therefore, it seems perfectly suited for the examination of degraded DNA. With the telomerase probe it was possible to obtain accurate results of the influence factors mentioned above. Based on the findings of this examination and on a case example, in which the poorly preserved remains of a baby were subjected to paternity testing, the possible uses of the telomerase assay are discussed. PMID- 16274075 TI - [Intramuscular bleeding in deaths from an internal cause: forensically significant differential diagnosis between external trauma and hemorrhages occurring during agony]. AB - Five cases of death from an internal cause are described in which bleeding into the muscles of the neck, the region of the shoulder girdle, the back of the thorax and partly the arms occurred during agony. As the cause of death was unclear at first and a violent crime could not be ruled out, a forensic autopsy was ordered in the course of which hemorrhages were detected which raised the differential diagnostic question whether extravasation might have been caused by an impact of blunt force. However, thorough investigation of the macroscopic and especially the microscopic structure of the lesions provided clear evidence that all the hemorrhages were due to internal causes. The pattern of these muscular lesions suggests intensified breathing with dyspnea leading to ruptures of the accessory respiratory muscles. Intramuscular bleedings of this type are a well known phenomenon in legal medicine in connection with death by drowning, but have not been described in the medicolegal literature in cases of natural death from an internal cause so far. PMID- 16274076 TI - Where has all the groundwater gone? PMID- 16274077 TI - Kinetics of removal of particulate chemical oxygen demand in the activated-sludge process. AB - The existing theories incorporated to state-of-the-art, activated-sludge consensus models indicate that the removal of particulate substrate from the liquid in the activated-sludge process is a two-step process: instantaneous enmeshment of particles and hydrolysis followed by oxidation. However, experimental observations indicate that the removal of particles is not instantaneous and needs a more accurate description. This removal process can actually be described as a three-step process: flocculation, hydrolysis, and oxidation. The principal objective of this research was to observe and model the kinetics of the removal of suspended particles and colloidal particles. A first order, particulate-removal expression, based on flocculation, accurately described the removal rates for supernatant suspended solids and colloidal chemical oxygen demand (COD). The rate of reaction for removal of colloidal COD was slow and comparable to that for soluble organic matter. PMID- 16274078 TI - Evaluation of ceramic and membrane diffusers under operating conditions with the dynamic offgas method. AB - The aeration systems of two full-scale, activated-sludge basins were compared during a period of three years, under the same operating conditions, using dynamic offgas testing. Only the material of the diffuser was different (membrane versus ceramic-tube diffusers). The investigation has shown that, although the membrane diffusers have higher initial standard-oxygen-transfer efficiency (alphaSOTE) and standard-aeration efficiency (alphaSAE), these decreased over time, while the alphaSAE of the ceramic diffusers started lower, but increased slightly over the whole period. A cost comparison makes clear how important it is to evaluate the aeration system under process conditions. The operating costs were the dominant factor (approximately 10x higher than capital costs), and operating costs were approximately 20% higher for membrane versus ceramic diffusers. The poor performance of the membrane-tube diffusers under process conditions could be explained on the basis of the actual alphaAE values in the basin, not the standardized values. PMID- 16274079 TI - Effects of reduced return activated sludge flows and volume on anaerobic zone performance for a septic wastewater biological phosphorus removal system. AB - Enhanced biological phosphorous removal (EBPR) performance was found to be adequate with reduced return-activated sludge (RAS) flows (50% of available RAS) to the anaerobic tank and smaller-than-typical anaerobic zone volume (1.08 hours hydraulic retention time [HRT]). Three identical parallel biological nutrient removal pilot plants were fed with strong, highly fermented (160 mg/L volatile fatty acids [VFAs]), domestic and industrial wastewater from a full-scale wastewater treatment facility. The pilot plants were operated at 100, 50, 40, and 25% RAS (percent of available RAS) flows to the anaerobic tank, with the remaining RAS to the anoxic tank. In addition, varying anaerobic HRT (1.08 and 1.5 hours) and increased hydraulic loading (35% increase) were examined. The study was divided into four phases, and the effect of these process variations on EBPR were studied by having one different variable between two identical systems. The most significant conclusion was that returning part of the RAS to the anaerobic zone did not decrease EBPR performance; instead, it changed the location of phosphorous release and uptake. Bringing less RAS to the anaerobic and more to the anoxic tank decreased anaerobic phosphorus release and increased anoxic phosphorus release (or decreased anoxic phosphorus uptake). Equally important is that, with VFA-rich influent wastewater, excessive anaerobic volume was shown to hurt overall phosphorus removal, even when it resulted in increased anaerobic phosphorus release. PMID- 16274080 TI - Factors affecting oxygen-transfer rates in headspace-gas respirometers. AB - Satisfactory measures of the biological-oxygen-uptake rate in headspace-gas respirometers can only be achieved if the rate of oxygen transfer from the headspace gas to liquid is greater than the rate of oxygen uptake by microorganisms. In the authors' study, factors potentially affecting oxygen transfer limitations in headspace-gas respirometers were evaluated quantitatively. Tests were conducted to measure maximum-oxygen-uptake rates by operating a respirometer under various test conditions. Analysis of respirometric data indicated that limiting oxygen-transfer rates were related to mixing intensity, length of magnetic stirring bar, volume of sample, and oxygen content in the headspace gas. A multivariable model was developed to describe the overall contribution of these factors to the limiting oxygen-transfer rate. This model should be useful for estimating maximum-oxygen-transfer rates for essentially all headspace-gas respirometers. PMID- 16274081 TI - Gamma irradiation of municipal sludge for safe disposal and agricultural use. AB - Gamma radiation was found to be an effective tool for hygienization of municipal wastewater sludge. The sludge received from the primary settling tank of a municipal wastewater treatment plant was gamma irradiated using a cobalt-60 source in a sludge hygienization research irradiator. The process parameters were adjusted to effectively eliminate coliform bacteria in the sludge and to prevent their regrowth. Irradiated sludge was found to be free of fecal coliform and could be directly disposed after drying in a landfill or used as manure. It could also be used as a medium for growth of Rhizobium sp for obtaining a bio fertilizer. PMID- 16274082 TI - Comparative assessment of municipal wastewater disposal methods in southeast Florida. AB - A comparative assessment of the risks of three effluent disposal alternatives currently available to wastewater utilities in Southeast Florida is presented in this paper. The alternatives are: deep well injection and ocean outfalls following secondary treatment, and surface water (canal) discharges following secondary wastewater treatment, filtration and nutrient removal. Water quality data, relative to disposal of wastewater treatment plant effluent were gathered, along with water quality data on the receiving waters, from utilities. Comparisons and conclusions regarding potential health concerns associated with the three disposal alternatives are presented. The results indicated that health risks associated with deep wells were generally lower than those of the other two alternatives. The proximity of injection wells to aquifer storage and recovery wells was a determining factor relative to injection well risk. Urban ecological risks were also indicated to be lower, though impacts of urban water use/reuse to the Everglades were not studied. Additional data collection and analysis were recommended to understand the effects of wastewater management on the cycling of water, nutrients and other constituents on southeast Florida. In particular, it was recommended that monitoring of effluents for nitrosamines and pharmaceutically active substances be implemented on a broad scale. PMID- 16274083 TI - Insights to false positive total cyanide measurements in wastewater plant effluents. AB - Many publicly owned treatment works in North America are exceeding permitted limits for total cyanide in their wastewater treatment effluents. A recently introduced rapid, segmented, flow-injection analysis procedure using UV digestion and amperometric detection of the membrane-separated cyanide was used to investigate the various scenarios by which elevated cyanide levels might be present in wastewater treatment plant effluent. A number of significant interferences can produce false positive bias during sample analysis with the traditional acid distillation technique, but are minimized or absent with the new analytical method. However, increased levels of cyanide were found in some chlorinated wastewaters compared to the levels before chlorination, suggesting a fast reaction mechanism associated with the disinfectant and some precursor in the wastewater. In particular, the contact of chlorine with nitrite in the presence of a carbon precursor appears to contribute to cyanide formation during wastewater treatment and sample handling. This paper explores the scenarios under which cyanide can form during wastewater treatment as well as those in which a false bias for total cyanide can be obtained during sample processing and provides guidance for appropriate sample handling, screening, and processing to ensure valid analytical results. PMID- 16274084 TI - An evaluation of the urban stormwater pollutant removal efficiency of catch basin inserts. AB - In a storm sewer system, the catch basin is the interface between surface runoff and the sewer. Responding to the need to improve the quality of stormwater from urban areas and transportation facilities, and spurred by Phase I and II Stormwater Rules from the U.S. Environmental Protection Agency, several companies market catch basin inserts as best management practices for urban water quality management. However, little data have been collected under controlled tests that indicate the pollutant removal efficiency of these inserts when the inflow is near what can be expected to occur in the field. A stormwater simulator was constructed to test inserts under controlled and replicable conditions. The inserts were tested for removal efficiency of total suspended solids (TSS) and total petroleum hydrocarbons (TPH) at an inflow rate of 757 to 814 L/min, with influent pollutant concentrations of 225 mg/L TSS and 30 mg/L TPH. These conditions are similar to stormwater runoff from small commercial sites in the southeastern United States. Results from the tests indicate that at the test flowrate and pollutant concentration, average TSS removal efficiencies ranged from 11 to 42% and, for TPH, the removal efficiency ranged from 10 to 19%. PMID- 16274085 TI - 2-Chlorobenzoate biodegradation by recombinant Burkholderia cepacia under hypoxic conditions in a membrane bioreactor. AB - The feasibility of applying bacterial hemoglobin technology to degrade 2 chlorobenzoate (2-CBA) through co-metabolism under hypoxic conditions in a membrane bioreactor (MBR) process has been studied in the laboratory. 2 chlorobenzoate removal and chloride release rates in the MBR system varied from 99 to 78% and 98 to 73%, respectively, depending on the operation conditions. Chemical oxygen demand (COD) removal efficiencies were more than 90% at food-to microorganism ratios ranging from 0.32 to 0.62 g/g/d, and the observed yield was 0.13 to 0.20 g biomass/g COD. The bacterial cell-floc size-distribution analysis showed that there is a significant change in bacterial floc size due to high shear stress during operation of the MBR. To characterize growth kinetics of Burkholderia cepacia strain dinitrotoluene, a mathematical model that describes co-metabolic oxidation of 2-CBA in an MBR has been developed. PMID- 16274086 TI - Three-dimensional, quantitative-structure-property-relationship study of aqueous solubility for phenylsulfonyl carboxylates using comparative-molecular-field analysis and comparative-molecular-similarity-indices analysis. AB - With both the comparative-molecular-field analysis (CoMFA) and the comparative molecular-similarity-indices analysis (CoMSIA), the paper describes two five component, three-dimensional, quantitative-structure-property-relationship (3D QSPR) models for the aqueous solubility logSw (Sw, mol x L(-1)) of 52 phenylsulfonyl carboxylates. Two models yield the leave-one-out cross-validated correlation coefficient q2 values 0.851 and 0.821, and the conventional correlation coefficient r2 values 0.963 and 0.929, respectively. The achievement of high q2 and r2 values of the CoMFA model indicates the significance of correlation of steric and electrostatic fields with the aqueous solubility. The key features in the CoMFA contour maps are critical to trace the important properties and gain insight to the solvation mechanism of tested compounds. The quality of CoMSIA model is slightly lower than that of CoMFA in terms of q2 and r2 values. Not requiring molecular superposition, CoMSIA may be faster than CoMFA in data processing. PMID- 16274087 TI - Temperature modeling in activated sludge systems: a case study. AB - A model of temperature dynamics was developed as part of a general model of activated-sludge reactors. Transport of heat was described by the one dimensional, advection-dispersion equation, with a source term based on a theoretical heat balance over the reactor. The model was compared to several reference models, including a tanks-in-series model and the dispersion model with heat components neglecting biochemical-energy inputs and other activated-sludge, heat-balance terms. All the models were tested under steady-state and dynamic conditions at a full-scale facility, the Rock Creek wastewater treatment plant in Hillsboro, Oregon, using meteorological data from a station located 16 km from the plant. The dispersion model and tanks-in-series model matched in situ temperature data with absolute-mean errors less than 0.1 degrees C. Neglecting biochemical-heat-energy inputs in the activated-sludge reactor underestimated temperatures by up to 0.5 degrees C. The biochemical-heat-energy inputs accounted for 30 to 40% of the total heat flux throughout the year. PMID- 16274088 TI - Scale-up considerations for a hollow-fiber-membrane bioreactor treating trichloroethylene-contaminated water. AB - Scale-up of a hollow-fiber-membrane (HFM) bioreactor treating trichloroethylene- (TCE-) contaminated water via co-metabolism with the methanotroph Methylosinus trichosporium OB3b PP358 was investigated through cost comparisons, bioreactor experiments, and mathematical modeling. Cost comparisons, based on a hypothetical treatment scenario of 568-L/min (150-gpm) flowrate with an influent TCE concentration of 100 microg/L, resulted in a configuration of treatment trains with two HFM modules in series and an overall annual cost of US dollar 0.36/m3 treated. Biological experiments were conducted with short lumen and shell residence times, 0.16 and 0.40 min, respectively, as a result of the cost comparisons. A new variable, specific transformation, was defined for characterizing the cometabolic transformation in continuous-flow systems, and values as large as 38.5 microg TCE/mg total suspended solids were sustainable for TCE treatment. Using mathematical modeling, HFM bioreactor system design was investigated, resulting in a five-step system design strategy to facilitate sizing of the unit processes. PMID- 16274089 TI - Continuous monitoring of conventional parameters to assess receiving water quality in support of combined sewer overflow abatement plans. AB - As part of its long-term control plan for combined sewer overflow (CSO) abatement, the city of Buffalo, New York, maintained a network of Hydrolab Datasondes (Hydrolab-Hach Company, Loveland, Colorado) to assess receiving water quality effects by continuously logging dissolved oxygen, pH, temperature, conductivity, and turbidity. Although the effect of individual CSOs could be visualized, turbidity levels entering the Buffalo River from the upper watershed often were greater than from CSO discharges. Turbidity data showed that the Buffalo River was a net-sediment sink. Low dissolved oxygen levels were observed in the summer during dry weather, baseflow, and watershed-wide storms and CSO events. Some CSOs did not produce dissolved oxygen sags in the receiving waters, but others did. This information, together with the sampling done for organic and inorganic contaminants, can aid the decisionmaking process when prioritizing outfalls for abatement work and provides a baseline against which receiving water quality improvements can be measured. PMID- 16274091 TI - One-stop cardiovascular diagnostic imaging (and radiation dose). AB - Kefer et al. indicate that "MSCT and MR are useful for the noninvasive detection of coronary artery stenoses on both a segmental and per vessel basis. Because neither technique is 100% accurate, these techniques are not ready yet to replace conventional coronary angiography." In summary, all things considered, a properly done cardiac catheterization with contrast angiography can really be a one-stop shop for diagnostic imaging and therapy of the cardiovascular system, and the radiation dose is acceptable. PMID- 16274090 TI - Phosphorus release in aerobic sludge digestion. AB - The objectives of this study are to examine the phosphorus release in aerobic sludge digestion and to better understand its governing mechanisms. In this study, phosphorus release was examined using the secondary sludge from both conventional and biological nutrient removal processes. The experiments were carried out at room temperature (22 +/- 2 degrees C), with or without automatic control of pH (4.5 to 7.8), and under three aeration schemes: fully aerobic (dissolved oxygen [DO] at 3 to 4 mg/L), low DO (0.2 to 0.8 mg/L), and cyclic (with alternate on/off aeration). The released phosphorus concentrations were 20 to 80 mg/L for the conventional sludge and 60 to 130 mg/L for the biophosphorus sludge. Higher phosphorus release also occurred at low pH (<6.0). As for the effect of DO, fully aerobic digestion caused higher phosphorus release than the low-DO and cyclic operations. For better understanding, the solid phosphorus in sludge was conceptually categorized into three forms: inorganic phosphorus precipitates, organic cellular phosphorus, and polyphosphate (poly-P) in polyphosphate-accumulating organisms. Dissolution of inorganic phosphorus precipitates is controlled by physical and chemical conditions, with pH being the most important in this study. Lowering the pH to 4 to 6 clearly promoted the release of inorganic phosphorus. Polyphosphate hydrolysis, on the other hand, was found to be regulated biologically (sensitive to occurrence of anaerobic conditions) and was insignificant in the glutaraldehyde-fixed sludge. Phosphorus release from organic phosphorus should correlate with the volatile solid (VS) digestion, which lyses the cells and frees the phosphorus covalently bonded with the organic matters. The amounts of phosphorus released per unit VS digested (deltaP/deltaVS) were therefore calculated for experiments with long periods of constant pH (to minimize interferences from dissolution/precipitation of inorganic phosphorus). The results suggested that some poly-P was hydrolyzed and released accompanying the aerobic VS digestion, but at rates far lower than those under anaerobic conditions. PMID- 16274092 TI - Anemia in heart failure--a concise review. AB - Heart failure affects 5 million persons in the United States, with 400,000 new cases occurring every year. Paradoxically, although advances in coronary angioplasty and effective drugs have increased survival post infarction, the myocardial damage and subsequent neurohormonal activation-induced remodeling causes significant morbidity years later in the form of heart failure. Angiotensin-converting enzyme inhibitors (ACEIs) and angiotensin receptor blockers (ARBs) together with beta blockers modify the neurohormonal activation associated with heart failure and are key treatments for improving cardiac function and survival. Anemia is a significant risk factor predicting morbidity and mortality in heart failure. This article describes the various etiologies of anemia in heart failure. Of particular importance is the fact that recent stem cell studies have shown that the drugs acting on the renin-angiotensin system inhibit erythropoiesis in vivo and may cause anemia in patients with both normal renal function and end-stage renal disease (ESRD). The role of angiotensin-II as an erythropoietic growth factor and ACE in facilitating erythropoiesis is described in this article. Anemia has been shown to be a modifiable risk factor and its treatment correlates with improvement in clinical outcomes. Thus, anemia, its etiology (especially the contribution of ACEIs and ARBs), physiologic and prognostic impact, and treatment in the setting of heart failure are critical areas for investigation. PMID- 16274093 TI - Relationship between endothelial function in the coronary and brachial arteries. AB - BACKGROUND: Endothelial dysfunction is the first step in the progression to atherosclerosis, but little is known regarding whether there is a correlation in endothelial function between the coronary and peripheral arteries. HYPOTHESIS: We investigated the relationship between coronary and peripheral endothelial function. METHODS: In 41 patients (mean age 63 years; 23 men, 18 women) with angiographically normal coronary arteries, changes in brachial artery diameter in response to hyperemic flow and sublingual nitroglycerin (NTG) were measured by high-resolution ultrasonography. During coronary angiography, acetylcholine (ACh, 3 and 30 microg/min) and NTG were infused into the left coronary ostium. The diameter of the coronary artery was quantitatively measured and coronary blood flow (CBF) was calculated by quantitative angiography and Doppler flow velocity measurements. Changes in these parameters in response to each drug infusion were expressed as the percent change from the baseline values. RESULTS: Flow-mediated dilation (FMD) of the brachial artery was 5.0 +/- 3.5% and correlated positively not only with the change in coronary diameter (ACh at 30 microg/min, r = 0.31, p < 0.05) but also with the change in CBF (ACh at 3 microg/min, r = 0.39, p < 0.05; ACh at 30 microg/min, r = 0.46, p < 0.01). Multivariate analysis demonstrated that FMD was one of the factors associated with the changes in coronary diameter and CBF. CONCLUSIONS: These results suggest that brachial endothelial function is associated with coronary endothelial function in patients with angiographically normal coronary arteries, suggesting that impairment of endothelial function may occur simultaneously in both coronary and peripheral arteries. PMID- 16274094 TI - Serum troponin I level after external electrical direct current synchronized cardioversion in patients with normal or reduced ejection fraction: no evidence of myocytes injury. AB - BACKGROUND: External electrical cardioversion (EEC) has been suggested as a cause of myocardial damage, but results from several previously published studies are conflicting. HYPOTHESIS: The purpose of the study was to evaluate myocardial electrical injury caused by EEC. METHODS: After elective EEC for atrial fibrillation (AF), cardiac troponin I (cTnI) was measured in 193 consecutive patients attending the Cardiology Department of the San Maurizio Hospital of Bolzano for elective EEC of AF over a period of 13 months. External electrical cardioversion was performed by one of the attending cardiologists with a synchronized monophasic defibrillator. Blood sample for cTnI was taken 18-20 h after EEC. RESULTS: Of 193 patients, 183 (95%) were successfully cardioverted. Mean number of shocks was 1.46 and the mean total energy discharged per procedure was 379.4 +/- 229.2 J. Cardiac troponin remained under the limit of confidence for all patients with a mean value of 0.017 +/- 0.021 mcrg/l. No correlation between total energy delivered and cTnI was found. In the subgroup of patients with low ejection fraction, none had elevated cTnI, and no difference in cTnI values between these and patients with an ejection fraction > 40% was found. CONCLUSIONS: The results of our analysis indicate that EEC caused no myocardial injury even in patients with low ejection fraction. PMID- 16274095 TI - Prognostic impact of demographic factors and clinical features on the mode of death in high-risk patients after myocardial infarction--a combined analysis from multicenter trials. AB - BACKGROUND: Contemporary information is lacking on the effect of demographic features and clinical features on the specific mode of mortality after myocardial infarction (MI) in the thrombolytic era. HYPOTHESIS: The aims of this study were (1) to examine the risk and trend of a different mode of mortality (i.e., all cause, arrhythmic, and nonarrhythmic cardiac mortality) in high-risk patients post MI with reduced left ventricular ejection fraction (LVEF) or ventricular arrhythmias; and (2) to assess the predictive value of demographic and clinical variables in the prediction of specific modes of death in high-risk patients post MI in the thrombolytic era. METHODS: In all, 3,431 patients receiving placebo (2,700 men, median age 64 +/- 11 years) from the EMIAT, CAMIAT, SWORD, TRACE, and DIAMOND-MI studies, with LVEF < 40% or ventricular arrhythmia were pooled. Risk factors for mortality among patients surviving > or = 45 days after MI up to 2 years were examined using Cox regression. Short-term survival (from onset of MI to Day 44 after MI) was also examined for TRACE and DIAMOND-MI, in which patients were recruited within 2 weeks of MI. RESULTS: After adjustment for treatment and study effects, age, previous MI/angina, increased heart rate, and higher New York Heart Association functional class increased the risk of all-cause, arrhythmic, and cardiac mortality. Male gender, history of hypertension, low baseline systolic blood pressure, and Q wave were predictive of all-cause and arrhythmic mortality, whereas diabetes was only predictive of all-cause mortality. Smoking habit and atrial fibrillation had no prognostic value. Similar parameters were also predictive of short-term mortality, but not identical. CONCLUSIONS: Our study has shown that in high-risk patients post MI, who have been preselected using LVEF or frequent ventricular premature beats, demographic and clinical features are powerful predictors of mortality in the thrombolytic era. We propose that demographic and clinical factors should be considered when designing risk stratification or survival studies, or when identifying high-risk patients for prophylactic implantable cardiodefibrillator therapy. PMID- 16274096 TI - Effectiveness of triple-site triggered atrial pacing for prevention of atrial fibrillation after coronary artery bypass graft surgery. AB - BACKGROUND: Recently, several temporary multisite pacing methods have been developed for prevention of postoperative atrial fibrillation (AF). HYPOTHESIS: In this study, we evaluated the effect of triple-site temporary triggered pacing in the AAT mode on the development of AF in patients undergoing coronary artery bypass graft (CABG) at high risk for developing postoperative AF. METHODS: A total of 70 patients undergoing CABG were randomly assigned either to pacing group (study group, n = 35 patients) or to no pacing group (control group, n = 35 patients). The external pacemaker was programmed to pace at the atrial triggered mode at a lower rate of 40 beats/min for 4 days. RESULTS: Atrial fibrillation, defined as lasting > 30 s, occurred in 4 patients (11.4%) in the study group and in 16 patients (45.7%) in the control group (p = 0.003). Sustained AF, defined as AF lasting > 10 min, also was observed less frequently in the study group than in the control group (11.6 vs. 37.1%, p = 0.024). Triple-site triggered atrial pacing was observed to reduce the incidence of AF by 75% and the incidence of sustained AF by 69%. CONCLUSIONS: We believe that multiple-site temporary pacing in the triggered mode is an effective way of preventing postoperative AF. This technique may be used especially in patients at high risk of developing AF. PMID- 16274098 TI - Anomalous coronary arteries: a familial clustering. AB - We present two brothers with anomalous origin of left coronary artery from the right sinus of Valsalva. Screening of other members of the family revealed no coronary artery anomalies. The likelihood of finding such a rare anomaly in two members of a family by chance alone is extremely small; extensive review of the literature found only three other reports of such familial association. PMID- 16274099 TI - Images in cardiology: acute anterior wall myocardial infarction in a young male with hypertrophic cardiomyopathy and myocardial bridging. PMID- 16274097 TI - Spironolactone improves diastolic function in the elderly. AB - BACKGROUND: Diastolic dysfunction is common in the elderly. Increased myocardial fibrosis, a major determinant of diastolic function, has been observed with advancing age. Spironolactone prevents age-related increases in myocardial fibrosis in old normotensive rats. HYPOTHESIS: Spironolactone, via its antifibrotic activity, can improve diastolic function in the elderly with isolated diastolic dysfunction. METHODS: The study was a prospective, double blind, randomized, placebo-controlled trial. Thirty elderly subjects between 60 and 85 years of age with isolated diastolic dysfunction and no contraindications for spironolactone were randomized to 25 mg/day of spironolactone or placebo for 4 months. Mitral E/A and deceleration time, plasma levels of carboxy-terminal of procollagen type I (PICP), and brain natriuretic peptide (BNP) were measured at baseline and at the end of 4 months. Plasma level of potassium was also monitored to prevent clinically significant hyperkalemia. RESULTS: There was no serious adverse event or clinically significant hyperkalemia in the spironolactone group. Compared with baseline values, spironolactone significantly improved mitral E/A ratio (0.71 +/- 0.08 vs. 0.84 +/- 0.19, p = 0.025) and deceleration time (285.5 +/- 73.1 vs. 230.0 +/- 54.7, p = 0.035). There were no significant differences in the magnitude of change in the levels of PICP and BNP between the two treatment groups. CONCLUSION: Spironolactone may improve diastolic function in the elderly. PMID- 16274100 TI - Allen B. Weisse. PMID- 16274101 TI - Induction of bladder lesion by terephthalic acid and its mechanism. AB - OBJECTIVE: To provide more information for rational evaluation of potential risks of terephthalic acid (TPA), we studied the effects of TPA on rats' bladders in 90 days after TPA exposure. METHODS: Sprague Dawley rats were subdivided into five groups, ingesting 0%, 0.04%, 0.2%, 1%, and 5% TPA respectively for a sub-chronic feeding study lasting for 90 days. Urine, serum and samples of brain, liver, lung, kidney, bladder, etc. were collected and analyzed. RESULTS: TPA ingesting decreased the value of urinary pH, and increased the contents of Ca2+, Zn2+, Mg2+, Na+, K+ in urine. The volume of 24 h urine was significantly increased in male rats in the 1% and 5% TPA groups. Urinary white sediment was found in both sexes, and its formation in male rats seemed more susceptible than that in female rats. Alpha 2u-globulin (AUG) in serum and urine of male rats was markedly increased in a dose-dependent manner. Fifteen cases of hyperplasia (simple or atypical) were determined in the 5% TPA ingesting group, 14/52 in male rats and 1/23 in female rats. Among them 3 male rats had no stone or calculus. Those with either bladder stones or hyperplasia were accompanied with urinary white sediments. CONCLUSION: White sediment accompanied with elevated urine AUG is the basis of TPA induced urolith formation, and is also associated with TPA induced bladder epithelial cell proliferation. It can act as an early biomarker for the potential toxic effect of TPA. PMID- 16274102 TI - Case fatality rate of severe acute respiratory syndromes in Beijing. AB - OBJECTIVE: To describe the case fatality rate of SARS in Beijing. METHODS: Data of SARS cases notified from Beijing Center for Disease Control and Prevention (BCDC) and supplemented by other channels were collected. The data were analyzed by rate calculation. RESULTS: The case fatality rate of SARS in Beijing was 7.66%, and had an ascending trend while the age of cases was getting older, and a descending trend while the epidemic development. The case fatality rate in Beijing was lower than that in other main epidemic countries or regions. CONCLUSIONS: The risk of death increases with the increment of age of SARS patients. Beijing is successful in controlling and treating SARS. PMID- 16274103 TI - Epidemiological features of severe acute respiratory syndrome in Beijing urban and suburb areas in 2003. AB - OBJECTIVE: To describe the epidemiologic features of an outbreak of severe acute respiratory syndrome (SARS) in urban and suburb areas in Beijing and to explore their differences between these two areas. METHODS: Data of SARS cases were collected from daily notification of China Ministry of Health and a database of infectious diseases was established by the Beijing Municipal Center for Disease Prevention and Control (BCDC). All the data were put into dataset files by Microsoft Excel-2000 and analyzed with SPSS version 10.0 software. RESULTS: The respective urban incidence and mortality rate were 29.06 and 2.21 per 100,000, while the case fatality rate was 7.62%. In contrast, the respective suburb incidence and mortality rate were 10.61 and 0.78 per 100,000, and the case fatality rate was 7.32%. No significant differences were found in demographic characteristics between the urban and suburb areas. CONCLUSION: Beijing urban area suffered a more serious SARS epidemic than the suburb area in 2003. PMID- 16274104 TI - Study on the total amount control of atmospheric pollutant based on GIS. AB - OBJECTIVE: To provide effective environmental management for total amount control of atmospheric pollutants. METHODS: An atmospheric diffusion model of sulfur dioxide on the surface of the earth was established and tested in Shantou of Guangdong Province on the basis of an overall assessment of regional natural environment, social economic state of development, pollution sources and atmospheric environmental quality. Compared with actual monitoring results in a studied region, simulation values fell within the range of two times of error and were evenly distributed in the two sides of the monitored values. Predicted with the largest emission model method, the largest emission of sulfur dioxide would be 54,279.792 tons per year in 2010. CONCLUSION: The mathematical model established and revised on the basis of GIS is more rational and suitable for the regional characteristics of total amount control of air pollutants. PMID- 16274105 TI - Effect of dermal exposure to paraphenylenediamine and linear alkylbenzene sulphonate in guinea pigs. AB - OBJECTIVE: To study the effects of paraphenylenediamine (PPD) and linear alkylbenzene sulphonate (LAS) alone and in combination on the skin. METHODS: Forty-eight guinea pigs were divided equally into 4 groups and exposed to PPD (4 mg/kg), LAS (12 mg/kg) and PPD (4 mg/kg) plus LAS (12 mg/kg) for 30 days. The biochemical parameters such as acid phosphatase, gtutathione-s-transferase, glutathione peroxidase, glutathione, lipid peroxidation and histamine contents in exposed skin were estimated. The histopathological examination of the exposed skin was also carried out. RESULTS: The skin enzymes, lipid peroxidation, and histamine increased while glutathione decreased in skin. The simultaneously exposed group showed additive toxic effects. The histopathological examination showed severe hyperkeratosis, thickening of collagen fibres and vacuolisation of epidermal cells in PPD plus LAS exposed skin. CONCLUSION: The findings of the present study suggest that simultaneous exposure to PPD and LAS has additive toxic effects. PMID- 16274107 TI - Preparation of monoclonal antibody and development of enzyme-linked immunosorbent assay specific for Escherichia coli O157 in foods. AB - OBJECTIVE: To prepare monoclonal antibodies (MAb) and antisera specific for Escherichia coli (E. coli) O157, and to develop a sandwich enzyme-linked immunosorbent assay (ELISA) to detect E. coli O157 in foods. METHODS: Spleen cells from BALB/c mice immunized with the somatic antigen of E. coli O157:H7 were fused with murine Sp2/0 myeloma cells. The hybridoma cell line specific for E. coli O157 was established after having been subcloned. Antisera specific for E. coli O157 was prepared by intravenous injection into New Zealand rabbits with a stain of E. coli O157:H7. The sandwich ELISA was developed with the polyclonal antibody as the capture antibody and the MAb 3A5 as the detection antibody. The inoculated ground poultry meat and pasteurized milk were tested to confirm efficiency of the method. RESULTS: MAb 3A5 specific for E. coli O157 and O113:H21 belonged to subtype IgM. The ascetic titers of the antibody was 1:1x10(6). No cross-reactivity of the MAb was observed with strains of Salmonella spp, Yersinia enterocolitica, Shigella dysenteriae, etc. The purified polyclonal antibody had a titer of 1:1x10(5) with E. coli O157. The detection limit of this sandwich ELISA was 10(3)-10(4) cfu E. coli O157/mL in pure culture with a high specificity, which was characterized by every non-O157 strain with negative response. With 10h enrichment procedure, E. coli O157:H7 recovered well from inoculated ground poultry meat and pasteurized milk at levels of 0.1 cfu/g and 0.1 cfu/mL. CONCLUSION: MAb 3A5 specific for E. coli O157 and O113:H21 can be produced by immunizing BALB/c mice with a strain of E. coli O157:H7. Then a sandwich ELISA can be developed with the polyclonal antibody as the capture antibody and the MAb 3A5 as the detection antibody. The method is proved to be a sensitive and specific technique to detect low number of E. coli O157 in food. PMID- 16274106 TI - Spatial genetic structure of two HIV-I-resistant polymorphisms (CCR2-64 I and SDF1-3'A) alleles in population of Shandong Province, China. AB - OBJECTIVE: To explore the spatial genetic structure of two HIV-I-resistant polymorphisms (CCR2-64 I and SDF1-3'A) alleles in the population of Shandong Province, China. METHODS: Using the techniques of spatial stratified sampling and spatial statistics, the spatial genetic structure of the locus (CCR2-64 I and SDF1-3'A), which was shown to be important co-receptor for HIV infection, was quantified from the populations of 36 sampled counties of Shandong Province, and a total of 3147 and 3172 samples were taken for testing CCR2-64I and SDF1-3'A respectively from individuals without known history of HIV-I infection and AIDS symptoms. RESULTS: There were significantly spatial genetic structures of the two alleles at different spatial distance classes on the scale of populations, but on the scale of individuals, no spatial structure was found in either the whole area of Shandong Province or the area of each sampled county. Although the change of frequencies of the two alleles with geographic locations in Shandong Province both showed gradual increase trends, their changing directions were inverse. The frequency of CCR2-64I allele gradually increased from the southwest to the northeast, while the frequency of SDF1-3'A allele gradually increased from the northeast to the southwest. However the RH to AIDS of combined types of their different genotypes did not represent obvious geographic diversity on the whole area of the Province. CONCLUSION: The frequency of allele usually has some spatial genetic structures or spatial autocorrelation with different spatial distance classes, but the genotypes of individuals have random distribution in the same geographic area. Evaluating spatial distribution of the genetic susceptibility of HIV (AIDS) to CCR2-64I and SDF1-3'A alleles, should focus on the frequencies of combined genotypes of CCR2 and SDF1 based on the two-locus genotypes of each individual rather than the frequencies of CCR2-64I and SDF1-3'A alleles. PMID- 16274108 TI - Elevated plasma tissue-type plasminogen activator (t-PA) and soluble thrombomodulin in patients suffering from severe acute respiratory syndrome (SARS) as a possible index for prognosis and treatment strategy. AB - OBJECTIVE: To detect the presence of endothelial injury in patients with severe acute respiratory syndrome (SARS) via enhanced levels of tissue-type plasminogen activator (t-PA) and soluble thrombomodulin (sTM). METHODS: Case patients were from Xuanwu Hospital (Capital University of Medical Sciences, Beijing, China), and all of them met clinical criteria for SARS. Healthy controls were some of the hospital employees. Endothelial injury bio-markers tPA and sTM were detected by commercial ELISA-methods. RESULTS: Classic plasma markers of endothelial injury, tPA and sTM significantly elevated in SARS patients in comparison to controls [t PA: 1.48 +/- 0.16 nmol/L versus 0.25 +/- 0.03 nmol/L (P<0.0001), and sTM: 0.26 +/ 0.06 nmol/L versus 0.14 +/- 0.02 nmol/L (P<0.05)]. The only patient who died had extremely high levels of these endothelial injury markers (t-PA: 2.77 nmol/L and sTM: 1.01 nmol/L). The likelihood ratio analysis indicated the excellent discriminating power for SARS at the optimal cut-point of 0.49 nmol/L for tPA and 0.20 nmol/L for sTM, respectively. Significant numerical correlations were found among these endothelial injury markers in SARS patients. The numerical coefficient of correlation Pearson r between t-PA and sTM was 0.5867 (P<0.05). CONCLUSION: Increased plasma concentrations of tPA and sTM in patients with SARS suggest the possibility of endothelial injury. SARS patients might need anticoagulant therapy or fibrinolytic therapy in order to reverse intraalveolar coagulation, microthrombi formation, alveolar and interstitial fibrin deposition. It may not only provide a useful treatment and prognostic index but also allow a further understanding of the pathological condition of the disease. PMID- 16274111 TI - Stand up and be counted. PMID- 16274109 TI - Development of a new high-throughput screening model for human high density lipoprotein receptor (CLA-1) agonists. AB - OBJECTIVE: To develop a new high-throughput screening model for human high density lipoprotein (HDL) receptor (CD36 and LIMPII analogous-1, CLA-1) agonists using CLA-1-expressing insect cells. METHODS: With the total RNA of human hepatoma cells BEL-7402 as template, the complementary DNA (cDNA) of CLA-1 was amplified by reverse transcription-polymerase chain reaction (RT-PCR). Bac-to-Bac baculovirus expression system was used to express CLA-1 in insect cells. CLA-1 cDNA was cloned downstream of polyhedrin promoter of Autographa californica nuclear polyhedrosis virus (AcNPV) into donor vector pFastBac1 and recombinant pFastBac1-CLA-1 was transformed into E. coli DH10Bac to transpose CLA-1 cDNA to bacmid DNA. Recombinant bacmid-CLA-1 was transfected into Spodoptera frugiperda Sf9 insect cells to produce recombinant baculovirus particles. Recombinant CLA-1 was expressed on the membrane of Sf9 cells infected with the recombinant baculoviruses. A series of parameters of DiI-lipoprotein binding assays of CLA-1 expressing Sf9 cells in 96-well plates were optimized. RESULTS: Western blot analysis and DiI-lipoprotein binding assays confirmed that CLA-1 expressed in insect cells had similar immunoreactivity and ligand binding activity as its native counterpart. A reliable and sensitive in vitro cell-based assay was established to assess the activity of CLA-1 and used to screen agonists from different sample libraries. CONCLUSION: Human HDL receptor CLA-1 was successfully expressed in Sf9 insect cells and a novel high-throughput screening model for CLA 1 agonists was developed. Utilization of this model allows us to identify potent and selective CLA-1 agonists which might possibly be used as therapeutics for atherosclerosis. PMID- 16274112 TI - Solving the athletic patient puzzle. PMID- 16274110 TI - Preparation of monoclonal antibodies against prion proteins with full-length hamster PrP. AB - OBJECTIVE: To prepare the PrP specific monoclonal antibodies (mAbs) that can be used for the detection of mammalian prions and study of pathogenesis of prion diseases. METHODS: Several BALB/c mice were immunized with recombinant hamster prion protein (HaPrP). Three hybridoma cell lines designated as B7, B9, and B10, secreting monoclonal antibodies against HaPrP, were established by hybridoma technique. The mAbs reactivities were evaluated with ELISA, Western blot, and immunohistochemistry. RESULTS: The mAbs produced by these cell lines reacted well with different recombinant hamster PrP proteins. Western blot analyses showed that mAbs B7 and B9 reacted with PrPSc from the scrapie-infected animals after proteinase K digestion with three glycosylated forms. The mAbs exhibited cross reactivity with various PrPC from several other mammalian species, including humans and cattles. Immunohistochemistry assays confirmed that mAbs B7 and B9 could recognize not only extracellular but also intracellular PrPsSc. CONCLUSION: The mAbs of prion protein are successfully generated by hybridoma technique and can be applied for the diagnosis of prion associated diseases. PMID- 16274113 TI - Oh, those aching backs! PMID- 16274114 TI - Strolling along. PMID- 16274115 TI - Biofeedback vs electrophysiology. PMID- 16274116 TI - Intimate assistance. PMID- 16274117 TI - In pursuit of (clinical) excellence. PMID- 16274118 TI - First you say you do, then you don't. PMID- 16274119 TI - Three issues to watch. PMID- 16274120 TI - [Repair and reconstruction of oral and maxillofacial defect--clinical analysis of 1973 cases]. AB - OBJECTIVE: To compare the reconstructive method of oral and maxillofacial defect with free tissue flaps. METHODS: The clinical materials were collected from 1 973 reconstructive cases between January 2000 and June 2004 and analyzed in terms of the distribution of age, gender, disease type, defect location, reconstructive method and the incidence of vascular crisis of free flaps as well as success rate of free flap respectively. SAS 6. 12 was adopted for statistical analysis. RESULTS: A total of 1973 reconstructive cases included 764 in middle age (>45 years to < or =60 years, 38.72%), 527 in old age (>60 years, 26.71%), 450 young adults (>28 years to < or =45 years, 22.81%), 187 in young age (>14 years to < or =28 years, 9.48%) and 45 children (< or =14 years, 2.28%). The ratio of male to female was 1.5 : 1. The ratio of benign to malignancy lesion was 1 : 1.94. The tongue defect accounted for 20.63%, followed by mandibular defect(17.38%), parotid defect(13.74%), buccal defect(12.72%), maxillary defect (8.16%), oral pharynx defect (7.60%), floor of mouth defect (5.68%) and others (14.09%). Vascular free flap transfers accounted for 45.82%(904), followed by axial flap(38.17%, 753), random flap (10.19%, 201), a vascularized bone graft (1.52%, 30) and others(4.30%, 85). The most frequently used flap was the forearm flap(594 cases), followed by the fibula free flap(143 cases) and the pedicled pectoralis major myocutaneous flap(369 cases); these three flaps accounted for 56.06% (1106/1973). In 47 free tissue flaps (5.20%) having vascular crisis, 30 were saved (63.83%). The success rate of total free tissue flaps was 98.19% (923/940). CONCLUSION: The majority of reconstructive cases of oral and maxillofacial defects is the middle aged and the old aged male patients with malignancy. The tongue defect accounts for about one fifth of all the cases. The vascularized free flap has a high success rate, so it is a main method for reconstruction of oral and maxillofacial defects. The forearm flap, the fibular free flap and the pedicled pectoralis major myocutaneous flap are the main management for repairing oral and maxillofacial defects. PMID- 16274121 TI - [Donor site choice for free flaps in head and neck reconstruction after tumor surgery]. AB - OBJECTIVE: To choose suitable free flaps for reconstructing head and neck defects caused by tumor resection. METHODS: A retrospective analyses was made in 86 cases of head and neck defects treated with four kinds of free flaps between January 1999 and January 2002. The head and neck defects were caused by tumor resection. The locations were oral cavity (n=32), hypopharynx (n=27), mandible (n=12), skull base (n=5), scalp and skin (n=6) and midface (n=4). The donor sites of free flaps included the rectus abdominis (n=32), anterolateral thigh (n=10), jejunum (n=25), fibula (n=11), latissimus dorsi (n=4), forearm (n=3) and scapula (n=1). The sizes of the cutaneous/musculocutaneous flaps ranged from 4 cm x 5 cm to 14 cm x 24 cm. The lengths of the fibula were 4-16 cm, of jejunum 9-20 cm. RESULTS: The overall free flap success rate was 92% (79/86). Of 32 oral cavity defects, 22 were reconstructed by rectus abdominis (69%) and 10 by anterolateral thigh flaps (31%). Of 27 hypopharyngeal defects, 25 were restored by jejunum flaps (93%). Eleven of 12 mandibular defects were reconstructed by fibula flaps (92%). Four of 5 defects of skull base were reconstructed by rectus abdominis flaps (80%). The free flaps of rectus abdominis, anterolateral thigh, jejunum and fibula were most frequently used, accounting for 91% (78/86) of all flaps in head and neck defect reconstruction. CONCLUSION: Although head and neck defects represent a complicated spectrum of subsites and loss, these four free flaps can manage most reconstruction problems. PMID- 16274122 TI - [Reconstruction of soft tissue defects in oral and maxillofacial regions after tumors surgery using cervical pedicle tissue flaps]. AB - OBJECTIVE: To report 4 methods of reconstructing soft tissue defects in oral and maxillofacial regions after tumors resection using cervical pedicle tissue flaps. METHODS: One hundred seventy-two soft tissue defects were repaired with cervical myocutaneous flaps after resection of oral and facial cancer (165 cases of squamous cell carcinoma and 7 cases of salivary carcinoma). The clinical stage of the tumors was stage I in 21 cases, stage II in 116 cases and stage III in 35 cases. Primary sites of the lesions were the tongue (59 cases), buccal mucosa (55 cases), lower gingiva (26 cases), floor of the mouth (25 cases), parotid gland (4 cases) and oropharynx (3 cases). Infrahyoid myocutaneous flaps were used in 60 cases, platysma flaps in 45 cases, sternocleidomastoid flaps in 59 cases and submental island flaps in 8 cases. The sizes of skin paddle ranged from 2.5 cm x 5.0 cm to 5.0 cm x 8.0 cm. RESULTS: Among 153 survival flaps, there were 55 infrahyoid myocutaneous flaps, 40 platysma flaps, 52 sternocleidomastoid flaps and 6 submental island flaps. There were 11 cases of total flap necrosis and 8 cases of partial flap necrosis. The success rates were 91.67% (55/60) for infrahyoid myocutaneous flap, 88.89% (40/45) for platysma flap, 88.14% (52/59) for sternocleidomastoid flap and 75% (6/8) for submental island flap. After a follow-up of 3 11 years (5.7 years on average) among 101 cases local recurrence in 18 cases, cervical recurrence in 4 cases, distance metastasis in 2 cases. The survival rate at 3 years were 83.17% (84/101). CONCLUSION: Cervical pedicle tissue flaps have clinical value in reconstruction of small and medium-sized soft tissue defects after resection of oral and maxillofacial tumors. PMID- 16274123 TI - [Platysma myocutaneous flaps in reconstruction of oral defect caused by tumor resection]. AB - OBJECTIVE: To study the method and feasibility of platysma myocutaneous flap in reconstruction of oral defects. METHODS: From March 2003 to June 2004, 15 patients with defects of tongue, floor of mouth, bucca, lateral pharynx and soft palate were treated with platysma myocutaneous flaps. There were 12 males and 3 females with an age range of 41-77 years. Defect was caused by resection of bucca (11 cases), tongue (1 case), floor of mouth (2 cases)and lateral pharyngeal and soft palate (1 case) squamous cell carcinoma. All patients were given primary cancer dissection and radical neck dissection. The defect sizes were from 3.0 cmX 3.5 cm to 7.0 cm X 4.0 cm. The sizes of the platysma myocutaneous flap were from 7.0 cm x 3.5 cm to 12.0 cm x 4.0 cm. RESULTS: All the patients were followed up for 6-12 months. In 15 patients, the flap survived, and the color and appearance of the flaps were normal. Mouth-opening was not limited. Eight patients had cutaneous fistula after 10-15 days of operation and were cured by secondary operation. No complication of ankyloglossia and tumor relapse occurred. The operation results were satisfactory. CONCLUSION: Platysma myocutaneous flap has generous blood supply, it is useful in reconstruction of oral tissue defects. PMID- 16274124 TI - [Repairing defects of tongue and mouth floor with submental island flap after tumor surgery]. AB - OBJECTIVE: To evaluate the clinical significance of submental island flap in repairing tongue defects. METHODS: Nine patients (6 men and 3 women) with tongue squamous cell carcinoma underwent subtotal or partial glossectomy, resection of mandible, radical neck dissection and immediate reconstruction of tongue defects with submental island flap. The age ranged from 48 years to 71 years, the lesion locations were right part of tongue (5 cases) and left part of tongue (4 cases). The defect sizes were 4.2 cm x 3.2 cm to 5.5 cm x 4.0 cm. The flap area ranged from 6.0 cm x 3.0 cm to 7.0 cm x 4.0 cm. The flap pedicle included submental artery in 8 cases and both submental artery and facial artery in 1 case. RESULTS: The submental island flap survived in 8 cases. Postoperative articulation and swallowing were investigated in all cases. The static shape of tongue after reconstruction with submental island flap was acceptable. The dynamic speech, swallowing and food transport function were well preformed. No complication occurred. Three patients were given radiotherapy 3 weeks after operation. CONCLUSION: It is simple and convenient to repair defects of tongue and oral floor with submental island flap. PMID- 16274125 TI - [Presurgical orthodontic treatment of complete unilateral cleft lip and palate in 100 infants]. AB - OBJECTIVE: To investigate the application and curative effect of naso-alveolar molding plate in the presurgical orthodontic treatment of complete unilateral cleft lip and palate in infants. METHODS: From January 2003 to March 2004, 100 infants with complete unilateral cleft lip and palate received presurgical orthopedics by using naso-alveolar molding plate. According to the age, 100 infants (aging 10 days to 3 months, including 60 males and 40 females) were divided into the test group (10 days to 1 month) and the control group (1-3 months). The widths of lip cleft and dentoalveolar cleft and the rate of satisfaction for nose wing were compared between 2 groups before treatment and after the treatment. RESULTS: The widths of cleft lip and dentoalveolar cleft before orthodontic (5.0 +/- 1.6 mm, 6.9 +/- 2.6 mm) and after orthodontics (4.1 +/- 2.7 mm, 6.4 +/- 2.9 mm)in the test group were less than those in the control group (7.5 3.1 mm, 12.5 +/- 4.0 mm and 8.3 +/- 3.0 mm, 10.8 +/- 2.6 mm), being statistically different (P < 0.05). The satisfactory rate of nose wing in the test group (86.3%) was significantly higher than that in the control group (62.4%, P < 0.05). CONCLUSION: The advantages of presurgical orthodontic treatment of complete unilateral cleft lip and palate in infants are as follows: repositioning the premaxillary segment, reducing the width of palate cleft, correcting the nasal deformities and facilitating surgical repair of cleft lip and palate. PMID- 16274126 TI - [Reconstruction of mandibular defect caused by resection of oral carcinoma with pectoralis major myocutaneous flap and ti-plate system]. AB - OBJECTIVE: To investigate the clinical effect of the pectoralis major myocutaneous flap and Ti-plate system in repairing mandibular defects caused by resection of oral carcinoma. METHODS: From November 2001 to February 2003, 32 patients with mandibular defect caused by resection of oral carcinoma were treated. Combined radical neck dissection with resection of gingival and mandible was performed on 11 patients with carcinoma of the lower gingival, combined radical neck dissection with glossectomy and mandibulectomy on 13 patients with carcinoma of tongue, combined radical neck dissection with resection of floor of mouth and mandible on 4 patients with carcinoma of floor of mouth, and combined radical neck dissection with resection of cheek and mandible on 4 patients with carcinoma of buccal mucosa, respectively. The defects of mandible were associated with soft-tissue component, the sizes of defect ranged from 5.5 cm x 7.6 cm to 8. 2 cm x 10.5 cm. The defects were reconstructed with 6 cm x 7 cm to 9 cm x 10 cm pectoralis major myocutaneous flaps and Ti-plate system. The effect was studied retrospectively. RESULTS: Thirty-two cases were followed for 2-19 months; 29 cases of flaps survived and 3 cases of flaps partly necrosed (10% or less of the skin paddle). The appearance of face was satisfactory in 27 patients, and slight deformity of face was observed in 5 patients. The occluding relation and masticatory function were recovered well. Opening mouth extents ranged from 2.7 cm to 3.4 cm. No temporomandibular arthrosis relating to operation was found in all cases. CONCLUSION: A combination of the pectoralis major myocutaneous flap and Ti-plate system is an ideal method for reconstruction of mandible defects associated with soft-tissue component after radical operation of oral carcinoma. PMID- 16274127 TI - [Evaluation of rigid internal fixation in mandibular reconstruction with autogenous bone]. AB - OBJECTIVE: To investigate the application of rigid internal fixation in mandibular reconstruction with autogenous bone and to evaluate its efficacy. METHODS: From January 1994 to May 2004, 98 patients with mandibular defect received mandibular reconstruction with autogenous bone by using rigid internal fixation. Seventy-two cases of benign tumor and 26 cases of malignant tumor were included. Four hundred and ono rigid fixation plates were inserted. The clinical results and X-ray films were analyzed and the healing processes were evaluated. The functional and aesthetic results of the mandibular reconstruction were also evaluated according to Lopez assessment system. RESULTS: After a follow-up of 1 to 3 years, 95 patients (96.9%)achieved successful effect. The forms and function of the mandibles were resumed. Eighty-one (82.7%) patients were satisfied with the results of operations. Thirteen patients (13.3%) achieved acceptable results. Four patients (4.1%) were dissatisfied with the results of operations. CONCLUSION: The rigid internal fixation is conductive to healing and remodeling of the transplant bone in mandibular reconstruction. PMID- 16274128 TI - [Clinical study on reconstruction of hemifacial atrophy with serratus anterior free-muscle flap]. AB - OBJECTIVE: To study the method of treating hemifacial atrophy with free serratus muscle flap. METHODS: Three patients diagnosed as having serious hemifacial atrophy was treated with free serratus muscle flap. The root of the flap was thoracodorsal artery and thoracodorsal vein, which was anastomosed with superficial temporal artery and vein, facial artery and vein, lingual artery and vein, and so on. During the operation, long thoracic nerve was dissected and anastomosed with facial nerve. The sizes of the flaps were 12 cm x 8 cm - 16 cm x 12 cm. RESULTS: All free-muscle flaps healed well after the transplant. The face and buccal area looked chubby and rounded. There were no obvious protuberance and discontentment on the buccal area. The shoulders of all patients moved well. The facial contour of the patients recovered well during the follow-up period (1-3 years). CONCLUSION: The method has a good result. The long-term effect needs further study. PMID- 16274129 TI - [Individual digital design and functional reconstruction of large mandibular defect with computer-aided design/computer aided manufacture technique]. AB - OBJECTIVE: To build up a new contour and functional reconstruction technique of mandibular defects with rapid prototyping and reverse engineering technique. METHODS: From April 2002 to August 2004, 4 cases of mandibular defects due to resection of large mandible lesion were treated. Of 4 patients, there were 3 females and 1 male, with an age range of 21-42 years, which underwent secondary operation and presented a deviation as mandibular movement. The opening-mouth extent was 1.8-2.5 cm (2.2 cm on average). The data of defects area were renewed with Mimics and Geomagic Studio software; and the titanium reconstructive frame was designed and manufactured with rapid prototyping technique. Defect were reconstructed by using CT digital data of patients. RESULTS: The CT data could be used by image software directly. The implant design could be completed by computer-aimed design (CAD) / computer-aided manufacture (CAM). The resin model and titanium frame were manufactured accurately by RP technique. Four patients achieved one stage healing. After a follow-up of 3 months to 2 years, large mandibular defect was reconstructed satisfactorily and the opening-mouth extent was 3.0-3.4 cm (3.2 cm on average). The occluding relation was normal. The implant denture was put on and the mastication function was good in 1 case. CONCLUSION: Individual design and repair of large mandibular defect with CAD/CAM techniques is worth extending application clinically. It is a simple and accurate method. PMID- 16274130 TI - [Combined vascularized iliac osteo-musculo-cutaneous flap with zygomatic implant anchorage in reconstructing 1 case of maxillary defect]. AB - OBJECTIVE: To reconstruct the maxillary defect by using free vascularized iliac osteo-musculo-cutaneous flap combined with immediate zygomatic implantation for early rehabilitation of maxillary contour and masticatory function. METHODS: In August 2003, the patient presented with deformity of left middle face (Brown II type defect) after subtotal maxillectomy. After hospitalization, a set of preoperative preparations were made, including spiral CT scanning, manufacture of nature size anatomical model and implantation protocol design. The maxillary defect was reconstructed with free vascularized iliac osteo-musculo-cutaneous flap combined with simultaneous insertion of one Branemark zygomatic implant and two general implants. Six months later the prosthesis were placed. RESULTS: The vascularized osteo-musculo-cutaneous flap survived, the osseointegration was observed between bone and implant 6 months later. The contour of face and palate was satisfactory, the normal occluding relation was gained. The average masticatory force of operative side was 76.3% of the normal side. No tumor recurrence was noticed during the follow-up of 14 months. CONCLUSION: It is a reliable method for functional reconstruction of maxillary defect via vascularized iliac osteo-musculo-cutaneous flap combined with immediate zygomatic implantation. PMID- 16274131 TI - [Fabrication of 3-dimensional skull model with rapid prototyping technique and its primary application in repairing one case of cranio-maxillo-facial trauma]. AB - OBJECTIVE: To investigate the methods of establishing 3-dimensional skull model using electron beam CT (EBCT) data rapid prototyping technique, and to discuss its application in repairing cranio-maxillo-facial trauma. METHODS: The data were obtained by EBCT continuous volumetric scanning with 1.0 mm slice at thickness. The data were transferred to work-station for 3-dimensional surface reconstruction by computer-aided design software and the images were saved as STL file. The data can be used to control a laser rapid-prototyping device (AFS 320QZ) to construct geometric model. The material for the model construction is a kind of laser-sensitive resin power, which will become a mass when scanned by laser beam. The design and simulation of operation can be done on the model. The image data were transferred to the device slice by slice. Thus a geometric model is constructed according to the image data by repeating this process. Preoperative analysis, surgery simulation and implant of bone defect could be done on this computer-aided manufactured 3D model. One case of cranio-maxillo facial bone defect resulting from trauma was reconstructed with this method. The EBCT scanning showed that the defect area was 4 cm x 6 cm. The nose was flat and deviated to left. RESULTS: The 3-dimensional skull was reconstructed with EBCT data and rapid prototyping technique. The model can display the structure of 3 dimensional anatomy and their relationship. The prefabricated implant by 3 dimensional model was well-matched with defect. The deformities of flat and deviated nose were corrected. The clinical result was satisfactory after a follow up of 17 months. CONCLUSION: The 3-dimensional model of skull can replicate the prototype of disease and play an important role in the diagnosis and simulation of operation for repairing cranio-maxillo-facial trauma. PMID- 16274132 TI - [One-stage reconstruction of large through-and-through defect of cheek with a combination of forehead skin flap and sternocleidomastoid island myocutaneous flap after cancer dissection]. AB - OBJECTIVE: To study the methods and results of a combination of forehead skin flap and sternocleidomastoid island myocutaneous flap in the reconstruction of large through-and-through defect of check. Methods One case of check cancer received ampliative resection and functional neck dissection. The defect area of the skin side was 9 cm x 7 cm, of the mucosa side 4.5 cm x 3.0 cm. The defect of the mucosa side was repaired with sternocleidomastoid island myocutaneous flap which blood supply was from thyroidea superior artery, occipitalis artery and carotis extra vein; of the skin side with forehead skin flap which blood supply was from temporalis superficialis artery and vein. The size of the sternocleidomastoid island myocutaneous flap was 5 cm x 3 cm, of the forehead skin flap 10 cm x 6 cm. RESULTS: Two flaps and the split survived after operation. One-stage healing was achieved. The patient was discharged from hospital 2 weeks after operation. The color and the quality were good. The tumor did not recur during follow-up of one year. The patient could take care of herself, and she lived normally in talk and diet. CONCLUSION: A combination of forehead skin flap and sternocleidomastoid island myocutaneous flap is a useful method to repair large through-and-through defect of cheek after cancer dissection. It is easy-to-operate and economical. PMID- 16274133 TI - [Experimental study on repair of critical-sized cranial defect by tissue engineered bone]. AB - OBJECTIVE: To evaluate repair of critical-sized cranial defect with tissue engineered bone fabricated by coral, bone mesenchymal stem cells (MSCs) and sustainedly released recombinant human bone morphogenetic protein 2 (rhBMP-2) by collagen. METHODS: Three scaffolds of rhBMP-2 + coral, collagen + rhBMP-2 + coral and MSCs+collagen+rhBMP-2+coral were fabricated. Forty New Zealand rabbits were made the models of critical-sized defects and divided into 5 groups according to different implants: group I, auto-ilium; group II, coral; group III, rhBMP 2+coral; group IV, collagen+rhBMP-2+coral; and group V , MSCs+collagen+rhBMP 2+coral. Repair of bone defect was evaluated after 8 and 16 weeks of implantation by gross observation, X-ray, HE staining and Masson's trichrome staining. RESULTS: Repair of bone defect in group V was similar to that in group I, and was better than that in group IV; and group III was worse. The gross appearance showed that defect region filled with bony tissue which had similar strength to adjacent bone and formed bone union with surrounding bone. The X-ray result displayed high radiopacity (80.45% +/- 2.52% in the 16th week). Histological observation showed new lamellar bone tissue and with few pore blank area. However, only transparent fibrous tissue filled the defect in group II. CONCLUSION: Collagen may be a suitable sustained release system for rhBMP-2. And MSCs may have important effect on enhancing repair of bone defect. Tissue engineered bone fabricated by MSCs+collagen+rhBMP-2+coral may be a useful material for bone defect repair. PMID- 16274134 TI - [Biological effect of WO-1 on human embryonic osteoblasts]. AB - OBJECTIVE: To investigate the effect of WO-1 on the proliferation and differentiation of human embryonic osteoblasts (HEO) and to provide research methods of bone tissue engineering. Methods HEO were isolated from periosteum and calvaria and then cultured in vitro. The dose-effect relationship between WO-1 concentration and biological effect of HEO was evaluated by growth curve and 3H TdR count. The effect of WO-1 on cell activity and proliferation was investigated by cloning efficiency, cell cycle analysis was determined by flow cytometer and morphological was examined through transmission electron microscope. Moreover, the effect of WO-1 on osteoblastic function was evaluated at protein and mRNA levels by ALP activity, 3H-proline incorporation, osteocalcin secretion (RIA) and mRNA expression of type I collagen and osteocalcin (RT-PCR). Results The proliferation of HEO was inhibited in high concentration of WO-1, while it was promoted in low concentration of WO-1. The optimal dose was 8 microg/ml, and there was dose-effect relationship in the certain range of WO-1 concentration (0.25 microg/ml to 8 microg/ml). In 8 microg/ml of WO-1, the cloning efficiency and cloning volume of HEO were increased, population doubling time was decreased. All indexes of osteoblastic function including ALP activity, type I collagen synthesis and osteocalcin secretion were increased, the more sufficed cell organs were observed under transmission electron microscope than control group (P < 0.05). CONCLUSION: WO-1 can promote the cell activity and proliferation of HEO cultured in vitro in low concentration, enhance the synthesis of extracellular matrix, such as type I collagen and osteocalcin, and accelerate the mineralization of osteoid. WO-1 can be used as a stimulant of proliferation and differentiation of HEO in the research of bone tissue engineering, which provide the theoretical basis in clinical application. PMID- 16274135 TI - [Primary study on histocompatibility of three kinds of collagen-chitosan porous scaffolds]. AB - OBJECTIVE: To construct three kinds of collagen-chitosan porous scaffolds with enhanced bio-stability and to investigate the histocompatibility of the scaffolds in vivo. METHODS: Collagen-chitosan porous scaffolds were fabricated by freeze drying method, cross-linked using dehydrothermal treatment and glutaraldehyde, respectively. The morphology of the uncross-linked scaffold (scaffold 1), dehydrothermal cross-linked scaffold (scaffold 2) and glutaraldehyde cross-linked scaffold (scaffold 3) was studied by scanning electron microscopy. Three kinds of scaffolds were embedded subcutaneously on dorsal surface of 12 rabbit ears. The general and local responses were recorded daily. The bio-stability and histocompatibility of the scaffolds were observed by using HE staining after 3, 7, 14 and 28 days of operation. RESULTS: The scaffolds had three-dimensional porous structures with a porosity of more than 90%, and possessed pore sizes of 120 +/- 10 microm, 80 +/- 15 microm and 170 +/- 20 microm, respectively. All experimental rabbits survived with good general condition during the study. All skin incisions healed well without obvious reactive red or swelling. Histological study showed that scaffold 1 was degraded rapidly with obvious inflammation. The degradation of scaffold 2 was slower than that of scaffold 1 and the inflammation of scaffold 2 was also milder than that of scaffold 1. Scaffold 3 possessed slow degradation property with slight inflammatory reaction, and rapid tissue regeneration. CONCLUSION: The collagen-chitosan porous scaffolds have three dimensional porous structures that are suitable for tissue regeneration. The bio stability and histocompatibility of the scaffolds are enhanced after cross linked. Glutaraldehyde cross-linked is better than dehydrothermal cross-linked, which can facilitate dermal tissue reconstruction. PMID- 16274136 TI - [Application of chitosan in cartilage tissue engineering]. AB - OBJECTIVE: To introduce the application of polymer material, chitosan, in the cartilage tissue engineering. METHODS: The recent original articles on the application of chitosan in cartilage tissue engineering were extensively reviewed. The biocompatibility and biodegradation characters of chitosan and its application were analysed. RESULTS: Chitosan has a high degree of biocompatibility and a favorable chondrogenic characteristic. It can support the maintenance of the phenotypic morphology of chondrocytes besides being used as a scaffold for cell growth. CONCLUSION: The perspective of the application of chitosan in cartilage tissue engineering is hopeful. PMID- 16274137 TI - [Research advance of ligament injury and ligament healing]. AB - OBJECTIVE: To introduce the research advance of the ligament injury and ligament healing. METHODS: Recent original articles related to such aspects of ligament were reviewed extensively. RESULTS: The ligament properties would be influenced when the situations of the biochemistry and biomechanics had changed. Injuries to ligaments induce a healing response that is characterized by scar formation. Graft could not recovery the ultrastructure, anatomy and biomechanics of the normal ligament. CONCLUSION: The healing ligament is weaker than normal one, and the graft could not reconstruct normal ligament at present. PMID- 16274138 TI - [Research advance of dynamic hip screw internal fixation in treatment of intertrochanteric fractures]. AB - OBJECTIVE: To investigate the development of dynamic hip screw (DHS) internal fixation in the treatment of intertrochanteric femur fractures. METHODS: The latest relevant articles were reviewed extensively, including biomechanics and clinical application research. RESULTS: DHS is the effective selection for the treatment of intertrochanteric femur fractures, especially by the innovation of DHS structure, operative manipulation. CONCLUSION: Treatment of intertrochanteric femur fractures with DHS is still a gold level, but strict selection of patients, proficiency operation and invasive manipulation are the most essential principles. PMID- 16274139 TI - Patient and clinician perspectives on quality of life. PMID- 16274140 TI - The effect of a machined collar on coronal hard tissue around titanium implants: a radiographic study in the canine mandible. AB - PURPOSE: The purpose of this study was to radiographically evaluate the effect of a machined titanium coronal collar on the marginal bone around 1-part endosseous dental implants placed at different heights relative to the bone crest. MATERIALS AND METHODS: Sixty dental implants were placed in edentulous spaces bilaterally in 5 foxhounds. Thirty test implants had a sandblasted, large-grit, dual acid etched surface (SLA) over the entire length of the implant. The other 30 implants (control) had a machined collar around the most coronal 1.8 mm of the implant; an SLA surface covered the remainder of the implant. Both control and test implants were placed at 3 distinct levels relative to the bone crest. Six implants (3 control and 3 test) were randomly placed side by side in each hemimandible. Radiographs were taken at placement (baseline) and monthly for 6 months postplacement using a standardized radiographic template. RESULTS: Fifty-eight of the implants integrated and were analyzed on each proximal surface. Bone loss occurred around all implants over the 6 months of the study. In general, implants placed with the top of the SLA surface above the bone crest had significantly less bone loss than implants with the top of the SLA surface placed flush with the bone level. Apically placed implants had greater bone loss than coronally placed implants. The magnitude of bone loss around paired control and test implants was approximately the same. DISCUSSION AND CONCLUSION: The least bone loss with each implant type was observed when the top of the implant was placed above the alveolar crest. When there was no machined collar, the least distance from the implant top to the bone crest (not, however, the least bone loss) was observed when the top of the implant was level with the bone crest. PMID- 16274141 TI - Clinical parameters associated with success and failure of single-tooth titanium plasma-sprayed cylindric implants under stricter criteria: a 5-year retrospective study. AB - PURPOSE: The purpose of this study was to determine the clinical parameters associated with long-term success and failure of single-tooth titanium plasma sprayed (TPS) cylindric implants. MATERIALS AND METHODS: Thirty-nine implants in 39 subjects were followed for 5 years. The following data were collected: subject age and gender, implant length, implant location, bone density, and implant position in relation to crestal bone. Assessments made at recall intervals included: Gingival Index (GI), probing depth, relative attachment level, and standardized radiographs. Failure was defined as a mean annual attachment loss rate (ALR) of > or = 0.25 mm after the first year of implant function. Between group differences were assessed nonparametrically using the Mann-Whitney and chi square tests. RESULTS: Nineteen implants were considered successes and 20 were considered failures with respective mean ALRs of 0.12 +/- 0.07 mm and 0.42 +/- 0.19 mm. The following factors were associated with success: longer implants (P < .001), lower GI (P < .001), higher bone density (P < .0001), and implant position at the crest or supracrestally (P <.0001). Age, gender, probing depth, and implant location were not related to outcome. CONCLUSIONS: A model using attachment loss as a parameter for success and failure has not been previously utilized. Longer implants, lower GI, higher bone density, and implant position at the crest or supracrestally were clinical factors associated with long-term success of single-tooth TPS cylindric implants in this patient population. PMID- 16274142 TI - Distraction osteogenesis versus autogenous onlay grafting. Part I: outcome of implant integration. AB - PURPOSE: The primary goal of this study was to compare bone-to-implant contact (BIC) in alveolar bone augmented by distraction osteogenesis with BIC in alveolar bone augmented by onlay iliac crest grafting. MATERIALS AND METHODS: Alveolar bone defects were created bilaterally in 5 American foxhounds, and after healing, bone augmentation was accomplished using distraction osteogenesis on 1 side of the jaw and onlay grafting on the other. Twelve weeks after consolidation, implants were placed in augmented and control sites. The animals were sacrificed and the jaws harvested for histologic analysis after an additional 8 weeks. RESULTS: The mean BIC was 54.7% +/- 14.6% for implants placed in distracted sites, 53.8% +/- 11.8% for sites where an onlay graft was used, and 51.2% +/- 14.4% for control sites. Significant differences in BIC were noted between experimental and control sites only at the apical third of the implant (19.8 +/- 1.8 for distracted sites; 15.5 +/- 1.5 for grafted sites; 8.0 +/- 0.5 for control sites; P < .05). DISCUSSION: The data showed that both distraction osteogenesis and onlay grafting produce sufficient bone for implant placement. There were no differences between procedures in regard to BIC after 8 weeks. CONCLUSION: These data suggest that both onlay grafting and vertical distraction are appropriate methods for bone augmentation prior to implant placement. PMID- 16274143 TI - Bone regeneration in standardized bone defects with autografts or bone substitutes in combination with platelet concentrate: a histologic and histomorphometric study in the mandibles of minipigs. AB - PURPOSE: To evaluate the effect of the addition of platelet concentrate (PC) to autografts or bone substitutes on bone regeneration in standardized bone defects. MATERIALS AND METHODS: Three standardized bone defects were prepared in both mandibular angles of 12 adult minipigs. The defects were grafted with autograft, anorganic bovine bone, or synthetic beta-tricalcium phosphate (beta-TCP). PC was added to only 1 side. The animals were divided into 4 groups, which were sacrificed at 4 different time points (1, 2, 4, and 8 weeks) for histologic and histomorphometric analysis. The concentrations of platelets and growth factors were measured to identify correlation to the histologic and histomorphometric results. RESULTS: No correlation was found between platelet count in whole blood and platelet count in PC (r(p) = 0.36). Furthermore, no correlation could be demonstrated between the platelet count of the PC and the concentrations of PDGF AB (r(p) = -0.27) and TGF-beta (r(p) = 0.34). There were no signs of a stimulating effect of PC on bone formation in combination with autografts or bone substitutes at any time point (P = .89). Addition of PC did not alter the pattern of graft degradation. DISCUSSION: The present study underlines the need for further investigation to identify the optimal concentrations of platelets and combinations of growth factors to achieve a predictable stimulatory effect on bone regeneration. One of the first steps to achieve this goal will be the development of a reliable method for the procurement of PC. CONCLUSION: PC had no impact on bone formation and graft degradation in standardized bone defects in the mandibles of minipigs. PMID- 16274144 TI - Bone density around titanium implants may benefit from smoking cessation: a histologic study in rats. AB - PURPOSE: This study tested the hypothesis that interruption of cigarette smoke inhalation (CSI) would reverse its impact on bone quality around implants. MATERIALS AND METHODS: Sixty-nine rats were assigned to 1 of 4 groups. Group 1 (n = 16) was the control group; group 2 experienced CSI for the duration of the study (150 days); group 3 experienced CSI for 83 days prior to implant placement, until 7 days prior to implant placement, when CSI ceased; and for group 4, CSI exposure was temporarily halted from 7 days before implantation to 21 days afterward. Bone density (the proportion of mineralized bone in a 500-microm-wide zone lateral to the implant) was calculated for each specimen (mean +/- SD). RESULTS: In the cortical bone, a slight difference in bone density was noted between the groups (97.66% +/- 3.69% for group 1, 98.30% +/- 0.95% for group 2, 98.83% +/- 0.73% for group 3, and 98.11% +/- 1.14 for group 4; P > .05). In contrast, continuous exposure to cigarette smoke (group 2) significantly decreased density in the cancellous bone in comparison to the other groups (25.69% +/- 9.41% for group 1, 18.08% +/- 6.0% for group 2, 25.46% +/- 5.42 for group 3, and 26.20% +/- 6.77% for group 4; P < .05), with no significant differences between groups 1, 3, and 4 (P > .05). DISCUSSION: The results support the concept that the effects of cigarette consumption on dental implants may be reversible, and therefore suggest that smokers may realize satisfactory outcomes if they cease smoking, even temporarily. CONCLUSION: In conclusion, smoking may affect bone quality around titanium implants in cancellous bone, and cessation could result in a return toward to the levels of the control group. PMID- 16274145 TI - Dimensional accuracy analysis of implant framework castings from 2 casting systems. AB - PURPOSE: To compare the dimensional accuracy of implant framework castings from an argon vacuum casting machine with those from a centrifugal casting machine. MATERIALS AND METHODS: Three 4 x 10-mm external hex-type implants (3i/Implant Innovations) were embedded in an acrylic resin block 7 mm apart, with a 2 mm offset of the middle implant. Eight reference points were marked on the implant collars. Twenty implant bar frameworks were waxed with UCLA abutments, invested with a ringless system, and subjected to the same thermal cycle. Ten wax patterns were cast in gold alloy using an oxygen-propane torch and centrifugal casting system; 10 were cast using an argon vacuum casting machine (KDF; Denken). The White 1-screw technique was applied after sequentially tightening the mesial and distal abutment screws to 10 Ncm. Fit of the implant framework castings was evaluated by measuring the marginal opening between the casting and implant at the reference points. These measurements were averaged and statistically compared for differences. RESULTS: The mean marginal openings at the most distant measuring locations from the tightened retaining screw at location 1 was between 44 to 48 microm for the centrifugal system compared to between 28 to 32 mm for KDF (P < .01). For screws tightened at location 3, the mean marginal openings at the most distant measuring locations were between 40 to 51 mm for the centrifugal system compared to between 27 to 29 microm for KDF (P < .01). DISCUSSION: In comparison with the centrifugal casting and oxygen-propane system, the argon vacuum system was more accurate and user friendly and less technique-sensitive. CONCLUSION: The argon vacuum casting machine tested produced more accurate, better fitting implant-supported prosthesis frameworks than a conventional centrifugal casting system. The "1-screw" method of evaluating casting fit was most effective when either of the prostheses' end screws were tightened. PMID- 16274146 TI - In vivo evaluation of hydroxyapatite coatings of different crystallinities. AB - PURPOSE: The influence of calcium phosphate (CaP) and hydroxyapatite (HA) crystallinity on bone-implant osseointegration is not well established. In this study, the effect of HA crystallinity and coating method on bone-implant osseointegration was investigated using a rat tibia model. MATERIALS AND METHODS: HA coatings 1 to 5 microm thick were produced using a supersonic particle acceleration (SPA) technology. The HA crystallinities used for this study were weight ratios of 30%, 50%, 70%, and 90%. A total of 128 HA-coated implants were placed into the tibiae of 64 male Sprague-Dawley rats. Bone-implant interfaces were evaluated using histology and push-out strength testing at 3 and 9 weeks after implantation. RESULTS: The 70% crystalline coatings exhibited significantly greater interfacial strength (5 implants/time point/treatment) than the 30%, 50%, and 90% crystalline coatings at 3 and 9 weeks following implantation. The implants with coatings of 70% crystallinity also had the greatest bone contact length. In addition, the HA coatings produced with SPA demonstrated greater interfacial strength and bone contact length than plasma-sprayed HA coatings (except for the HA coating with 30% crystallinity). DISCUSSION: HA coatings of different crystallinities exhibited different dissolution and re-precipitation properties which may enhance early bone formation and bone bonding. CONCLUSIONS: This study suggested that coating crystallinity and coating methods can influence the bone-implant interface. PMID- 16274147 TI - In vitro evaluation of the implant-abutment bacterial seal: the locking taper system. AB - PURPOSE: To test in vitro whether the seal provided by the locking taper used in the implant-abutment connection was capable of preventing the invasion of oral microorganisms. MATERIALS AND METHODS: Twenty-five wide-body implants (5 x 11 mm) and 25 abutments were divided into 2 groups for a 2-phase experiment. The first phase tested the ability of the seal to shield the implant well from outside bacteria; the second phase tested the ability of the seal to prevent bacteria present in the implant well from seeping out. For phase 1, 10 implant-abutment units were immersed in a bacterial broth for 24 hours. The abutments were then separated from the implants and bacterial presence was evaluated using scanning electron microscopy. In phase 2, the tested abutments were inoculated with a droplet of soft agar bacterial gel and assembled with the implant. These units were incubated in a sterile nutrient broth for 72 hours, sampled, and plated to assess bacterial presence. RESULTS: In phase 1, no bacteria were detected in any of the implant wells. In phase 2, no bacteria were detected in the nutrient broth or on the agar plates at 72 hours. DISCUSSION: In implants where a microgap is present, microbial leakage could lead to inflammation and bone loss; thus, it is important to minimize bacterial presence in and around the the implant-abutment junction. CONCLUSION: The seal provided by the locking taper design has been demonstrated to be hermetic with regard to bacterial invasion in vitro. PMID- 16274148 TI - Implant prosthodontic rehabilitation of fibula free-flap reconstructed mandibles: a Memorial Sloan-Kettering Cancer Center review of prognostic factors and implant outcomes. AB - PURPOSE: This study aimed to estimate the cumulative survival rates (CSRs) of implants placed in reconstructed mandibles and to identify prognostic factors that may influence implant survival. MATERIALS AND METHODS: The charts of 24 patients (10 male, 14 female) who had undergone mandibular resection and reconstruction with fibula free-flaps treated with implant-supported prostheses from April 1986 through December 2001 were reviewed. Information on demographics, surgical characteristics, treatment modalities, dentition, implant parameters, prostheses, and hyperbaric oxygen therapy (HBO) was gathered. Kaplan-Meier survival estimates were generated for the 100 implants that satisfied the inclusion criteria. Multivariate Cox proportional hazards regression models accounting for correlated implants within subjects were developed to identify prognostic factors for implant survival. RESULTS: Ninteen implants had been placed in native mandible (3 in irradiated bone) and 81 in fibula bone flap. Six implants failed during the follow-up period (mean 51.7 months). The overall 5- and 10-year CSRs were 97.0% and 79.9%, respectively. In the univariate analysis, variables associated with implant survival were age, gender, chemotherapy, radiation therapy, HBO, irradiated bone, implant diameter, xerostomia, trismus, opposing dentition, and type of prosthesis. At 5 years, the CSR of implants in patients with HBO was 86.7%; HBO was statistically associated with an increased risk for implant failure (P = .005, hazard ratio = 19.79, 95% CI: 2.42 to 161.71). DISCUSSION: The CSR was lower when implants were placed in a previously irradiated mandible. There is still a lack of reliable clinical evidence to support the effectiveness of HBO in these patients. CONCLUSIONS: A high survival rate was demonstrated for implants placed in fibula free-flap reconstructed mandibles. The finding that HBO was a risk factor can probably be attributed to the small sample size; further study is needed in this patient population. PMID- 16274149 TI - Stability measurements of 1-stage implants in the maxilla by means of resonance frequency analysis: a pilot study. AB - PURPOSE: The objective of the present study was to determine standard Implant Stability Quotient (ISQ) values for apparently successfully osseointegrated 1 stage implants in the maxilla. MATERIALS AND METHODS: To measure implant stability, resonance frequency analysis (RFA) was performed in 35 patients (18 women, 17 men) with a total of 120 maxillary ITI implants. Based on the time interval between implant placement and measurements, the ISQ values of anterior and posterior implants were divided into subgroups: unloaded (n = 41), loaded < or = 12 months (n = 31), and loaded > 1 year (n = 48). Statistical analysis was performed using a mixed-effects model with the variables lading, implant location, and gender as fixed effects. RESULTS: The mean ISQ of all measured implants was 52.5 +/- 7.9 (range 40 to 68). Statistical analysis showed no significant differences in ISQ values between the 3 tested time intervals: unloaded (48.8 +/- 3.6), loaded < or = 12 months (54.1 +/- 7.0), and loaded > 1 year (53.1 +/- 9.5). Neither for the location in the jaw nor for bone quality (assessed using radiographs) could a significant difference be found. Gender was the only parameter which was found to be significant (P < .003); on average, men showed higher implant stability than women (56.3 +/- 6.6 versus 48.7 +/- 7.4). DISCUSSION: Standard values for osseointegrated maxillary ITI implants exhibited an individual range. Single RFA measurements of an implant do not allow assessment of its current status or prediction of its performance. Repeated measurements over a longer time period would be necessary. CONCLUSIONS: No significant differences in ISQ values were found between implants with regard to loading period or location in the jaw. Postmenopausal women exhibited significantly lower ISQ values compared to men of the same age group. PMID- 16274150 TI - Interproximal papilla levels following early versus delayed placement of single tooth implants: a controlled clinical trial. AB - PURPOSE: The aim of this study was to evaluate interproximal papillae and clinical crown height following the placement of single-tooth implants according to early and delayed protocols. MATERIALS AND METHODS: Forty-five patients were randomly allocated to either the "early" group or the "delayed" group. They were treated with a single-tooth acid-etched Osseotite implant in the maxillary or mandibular anterior or premolar region an average of 10 days (in the case of early placement) or 3 months (in the case of delayed placement) following tooth extraction. Interproximal papilla dimensions and clinical crown height were evaluated using a score index in 39 patients who attended a follow-up visit 16 to 18 months after prosthesis delivery. The patients were evaluated in photographs taken 1 week after crown placement (baseline) and approximately 1.5 years after crown placement (follow-up). RESULTS: It was demonstrated by logistic regression the risk of presenting no papilla or a negative papilla was 7 times greater at baseline for delayed cases than for early cases (33% versus 8%). However, the soft tissue fill in the proximal spaces improved significantly from baseline to the 1.5-year follow-up in both groups, with no significant difference between the groups found at follow-up. The papilla height almost 2 years after implant placement was inversely correlated with patient age. The clinical crown height was acceptable in significantly more cases in the early group than in the delayed group at follow-up. Half of the crowns in the delayed group exhibited an inappropriate height; of these, almost two thirds were assessed to be too short. DISCUSSION AND CONCLUSION: Early placement of single-tooth implants may be preferable to delayed implant placement technique in terms of early generation of interproximal papillae and the achievement of an appropriate clinical crown height, but no difference in papilla dimensions was seen at 1.5 years after seating of the implant crown. PMID- 16274151 TI - Effect of mandibular ridge height on patients' perceptions with mandibular conventional and implant-assisted overdentures. AB - PURPOSE: This study assessed the impact of mandibular ridge height on patients' perceptions of dentures following treatment with a mandibular conventional denture (CD) or an implant-assisted overdenture (IOD). MATERIALS AND METHODS: Evaluation of patient satisfaction in 63 participants was made with original complete dentures and 6 months after treatment completion with new dentures. Twenty-five patients received a new mandibular CD and 38 received a new mandibular IOD. The subjects were divided into 3 subgroups according to ridge height (low, moderate, or high). Two questionnaires with categorical responses were administered. Questionnaire 1 had 13 questions to determine patients' assessment of their original dentures at entry and of their study dentures at 6 months after treatment completion. Questionnaire 2, which was given at 6 months after treatment completion, had 11 questions assessing the change perceived by patients with new dentures compared to their original dentures. RESULTS: No significant differences between the 2 groups were found for most of the variables in Questionnaire 1 at either time point or in regard to the difference between time points. The retrospective questionnaire 2 showed the IOD group to have significantly better perceptions than the CD group for improvement in chewing comfort, ability to eat hard foods, eating enjoyment, and denture security. The only effect of ridge height was an interaction with denture treatment for eating enjoyment, where mean improvement with the study denture was significantly less for the moderate ridge height group with the CD. CONCLUSION: The results indicate that patients in all ridge height groups had similar improvement in perceptions of dentures following treatment with either a mandibular CD or IOD and that these perceptions were not dependent on the bone height of the mandibular ridge. PMID- 16274152 TI - Correlation between placement torque and survival of single-tooth implants. AB - PURPOSE: This study evaluated the survival parameters of single-tooth implants through clinical and radiographic analysis. MATERIALS AND METHODS: Implants were restored within a 24-hour period with a provisional crown designed to receive an occlusal masticatory load. This approach was compared to implants restored after a healing period (the control group). Forty-six implants were placed in 23 patients who were each treated with 2 Frialit-2 implants placed in sites between the second premolar in the maxilla or mandible. The manufacturer's recommended formal surgical procedure was followed, and primary stability was standardized with a minimum insertion torque of 20 Ncm. The sites were randomly selected, and the clinical and radiographic parameters were standardized with individual templates. RESULTS: Data were collected at 24 h, and at 1, 3, 6, 12, 18, and 24 months. The experimental group included 10 failed implants; 9 of the failed implants had been placed with an insertion torque of 20 Ncm. One implant from the control group failed during the 24-month follow-up period. The survival rate was independent of implant length, site position, and bone quality and quantity. Relative risk for implant failure was associated with insertion torque (relative risk 0.79 [CI: 0.66-0.930]; Cox regression) (P < or = .007), in the experimental group but was not significant for those in the control group (ie, implants placed after a healing period; relative risk 0.78 [CI: 0.34-1.78]; Cox regression) (P < or = .057). To achieve osseointegration, it was found that an insertion torque above 32 Ncm was necessary (chi2= 15.68; P < or = .004). DISCUSSION: A careful evaluation is necessary for a better understanding of the survival rates of immediately loaded implants. In this study, insertion torque was associated with the potential for risk, which can be decreased by 20% per 9.8 Ncm added. CONCLUSION: Given these results, and considering the number of patients treated, immediate provisional crowns should only be proposed with early loading if an appropriate initial insertion torque has been applied. PMID- 16274153 TI - Palatal core graft for alveolar reconstruction: a new donor site. AB - PURPOSE: A simple technique is described for alveolar reconstruction in cases where resorption has already occurred. The palatal core graft allows immediate regeneration of vertical, horizontal, and combined defects at the alveolus with minimal donor site morbidity and time consumption. MATERIALS AND METHODS: Seventeen patients (10 female and 7 male) were treated over a 1-year period. Bone core grafts from the palatal vault were harvested with a trephine and used for alveolar reconstruction in the esthetic zone. Patient age and gender, type and location of the defect, size of the graft, simultaneous tooth removal, simultaneous implant placement, need for soft tissue coverage, and postoperative complications were recorded. RESULTS: Mean length of the grafts was 12.5 mm (range 9 to 14 mm). Mean width was 7.3 mm (range 5 to 9 mm). In 11 cases, simultaneous tooth removal was performed and a connective tissue flap with posterior pedicles was used. Patients did not experience discomfort in the donor area. Sixteen cases were successfully restored with single implants and crowns DISCUSSION: Material for osseous reconstruction should ideally be autologous and easy to harvest and provoke minimal morbidity. The palatal core graft compares favorably with previously described techniques and donor sites for alveolar reconstruction. CONCLUSION: In this preliminary report, experience with a new technique for alveolar reconstruction is presented. The "palatal core graft" for alveolar reconstruction is effective and easy to harvest and has low donor site morbidity, allowing 3-dimensional restoration of alveolar defects. PMID- 16274154 TI - Distribution of the maxillary artery as it relates to sinus floor augmentation. AB - PURPOSE: Knowledge of the blood supply to the sinus is of importance in sinus augmentation, both as it pertains to vascularization of the sinus graft and as its location relates to the position of the required lateral osteotomy. The purpose of this study was to investigate the distribution of the endosseous branches of the maxillary artery in the area of the proposed lateral window. MATERIALS AND METHODS: Fifty computerized tomographic (CT) scans from 625 available patients undergoing sinus augmentation surgery at the New York University Department of Implant Dentistry were chosen at random for evaluation. In those cases where the maxillary artery could be identified, measurements were taken to determine the distance between the alveolar crest and the lower border of the vessel. RESULTS: The vessel was radiographically identified in 51.4% of right sinuses and 54.3% of left sinuses in the 50 CT scans. The average height of the artery from the alveolar crest was 16 mm (+/- 3.5 mm). DISCUSSION: and CONCLUSIONS: Because of its location, the intraosseous artery has the potential to cause bleeding complications in approximately 20% of normally positioned lateral window osteotomies. Although a previous anatomic study on cadavers identified the vessel in 100% of the specimens, it could only be visualized in 53% of the CT scans in the present series. PMID- 16274155 TI - Zygomatic implants using the sinus slot technique: clinical report of a patient series. AB - PURPOSE: The management of 5 patients with extreme maxillary atrophy and treatment consisting of maxillary fixed prostheses supported by conventional implants and zygomatic fixations positioned according to the sinus slot technique is reported. MATERIALS AND METHODS: A total of 16 conventional implants were placed, together with 2 pterygoid implants and 10 zygomatic fixations. In 2 cases zygomatic fixation could not be performed on the alveolar ridge, thus requiring palatal displacement. One patient presented nasogenian ecchymosis. The fixed rehabilitations were either screwed or cemented after 5 to 6 months. RESULTS: Follow-up from implantation lasted 12 to 18 months, during which the prostheses and implants remained stable and in function. DISCUSSION: The placement of zygomatic fixations based on the sinus slot technique offers advantages over the conventional technique, though extreme atrophy of the alveolar processes does not allow fixation at the supracrestal level, and complications may develop. CONCLUSION: While zygomatic fixation is a valid alternative for treating the atrophic jaw, long-term studies are required to confirm its efficacy. PMID- 16274156 TI - Clinical evaluation of dental implants with surfaces roughened by anodic oxidation, dual acid-etched implants, and machined implants. AB - PURPOSE: The purpose of this study was the clinical and radiographic comparison of dental implants with surfaces roughened by anodic oxidation (TiUnite), dual acid-etched implants (Osseotite), and machined implants. MATERIALS AND METHODS: Seventy-four patients (mean age, 52.8 +/- 14.2 years; range, 23 to 80 years; 41 men and 33 women) received 198 dental implants-58 TiUnite implants (25 patients), 52 Osseotite implants (27 patients), and 88 machined implants (22 patients). Clinical measurements and radiographs were evaluated at the time of surgery, at the restorative phase, and 2 years postloading. To account for statistical correlation among multiple implants in the same subject, a "per patient" mode of analysis was conducted. A 1-way analysis of variance of bone loss was conducted by type of implant as well by area of the mouth. In addition, differences in mean bone loss were tested for bone density category, gender, and smoking status using Student t tests. RESULTS: Eighteen TiUnite implants (31.0%) were placed in the maxilla and 40 (69.0%) in the mandible. The Osseotite group included 29 maxillary implants (55.8%) and 23 mandibular implants (44.2%). The machined group included 49 maxillary implants (55.7%) and 39 mandibular implants (44.3%). All 198 implants were considered radiographically and clinically successful. No mobility, signs of infection, or inflammation were detected. DISCUSSION: Implant size, location, bone quality, gender, age, and smoking did not influence the comparative clinical outcomes of the 3 groups (P > .05). A trend toward greater coronal bone loss in the TiUnite group was detected. CONCLUSION: Within the limitations of the present study, TiUnite, Osseotite, and machined dental implants had similar short-term clinical outcomes. No statistically significant differences in bone loss could be detected among implant groups or among the different regions of the oral cavity. The present data underlined the significance of surgical and prosthetic treatment planning. PMID- 16274158 TI - For bird flu, good PR equals bad medicine. PMID- 16274157 TI - Surgical management of the partially edentulous atrophic mandibular ridge using a modified sandwich osteotomy: a case report. AB - Loss of posterior mandibular teeth resulting in insufficient posterior vertical bone height can pose a problem in implant therapy. In the case reported, a modified sandwich osteotomy was used to solve this problem. PMID- 16274159 TI - The coming storm over a cancer vaccine. PMID- 16274160 TI - Diet stocks for the post-Atkins age. PMID- 16274161 TI - Made for the shade: the Hawaii Skin Cancer Coalition celebrates its ten year history. PMID- 16274162 TI - The obstetrician's role in newborn metabolic screening: a physician survey. AB - Seventy-seven of 247 (31.2%) Hawaii obstetricians completed a questionnaire about newborn metabolic screening (NBMS). Only 13.0% correctly answered the knowledge questions, and 81.8% reported that they did not discuss NBMS with patients. Of the minority of obstetricians who discuss NBMS, only 25.0% correctly answered the knowledge questions. The study results revealed that obstetricians need to receive appropriate NBMS education and be encouraged to discuss NBMS with their patients. PMID- 16274163 TI - Overweight among Kauai first-graders. AB - This 2003 study examined pre-school physical exam data on student health records of Kauai public school first-graders and found that 14.5% were at risk for overweight (between > or =85th and 94th percentile BMI-for-age), and 21.5% were overweight (>95 percentile BMI-for-age). The prevalence of overweight among Kauai students was twice that reported nationally for the same age group. PMID- 16274164 TI - A case report of adult croup: a new old problem. AB - Larnygeotracheobronchitis is a common disease of childhood most often associated with infection by parainfluenza virus. Although rare, adult cases have been reported. Adult croup presents similarly to the childhood version; however, the severity is usually much worse, often leading to hospitalization. Presented is the case of an 80-year-old woman with croup, which is the twelfth reported in the English literature. PMID- 16274165 TI - Humanism: the greatest technological advance in medicine. PMID- 16274166 TI - Dietary changes in Korean American immigrants: implications for chronic disease risk. PMID- 16274167 TI - Is ICE as simple as it sounds? EMS Providers question the concept. PMID- 16274168 TI - An instant to decide, a lifetime to regret: the aftermath of ambulance accidents. PMID- 16274170 TI - Cheap, fast and easy. PMID- 16274169 TI - Late again: coping with those schedule headaches. PMID- 16274171 TI - "Is there someone on board?". PMID- 16274172 TI - Resuscitation in 2005: new ways to optimize manual CPR. PMID- 16274173 TI - Prehospital heart sounds. PMID- 16274174 TI - Cardiac case study. 50-year-old male with chest pressure. AB - This case study represented a patient with a relatively uncomplicated myocardial infarction that, after prompt prehospital care and transport, was successfully halted in the emergency department with fibrinolytic therapy. This patient was provided excellent care in the prehospital setting because the paramedic and his EMT-B partner worked together effectively as a team. Although ECG monitoring, IV therapy and medication administration are beyond the usual scope of practice of an EMTB, many EMS systems are training their EMT-Bs to assist with these important procedures and interventions. This involves preparing IV equipment and supplies, applying the cardiac monitor, and recognizing and handling the various paramedic medications. This enhanced role of the EMT-B allows the paramedic to perform a more focused and careful patient assessment. A cohesive working relationship between BLS and ALS personnel is absolutely crucial to the outcome of the patient. Although each level of prehospital provider possesses a different knowledge of pathophysiology and patient management, it is the combined contributions and efforts of each provider that will afford patients the high quality of care they deserve. PMID- 16274175 TI - Unusual resuscitation situations: environmental emergencies. PMID- 16274176 TI - Congestive heart failure and noninvasive positive pressure ventilation. PMID- 16274177 TI - Marfan Syndrome, aortic dissection and the EMS provider. PMID- 16274178 TI - A disaster doesn't have to be a disaster. An evidence-based triage method that "takes the guesswork out of triage". PMID- 16274179 TI - Quality at its best. PMID- 16274180 TI - Budgeting: an analysis of the basic key concepts. AB - The process of budgeting can be the most important tool available to an EMS department management team. The person(s) responsible for the oversight of the department should have a working knowledge of both the budget numbers themselves and, more important, the process of working through a budgetary cycle. This should include members of the management team who do not have direct responsibility for preparation and presentation of the budget, as they will often be the persons who assist the preparer in budgetary maintenance over its life cycle. A working knowledge of basic financial concepts by all members of the management team is paramount to the continued viability and success of any proactive EMS department. PMID- 16274181 TI - Temp industry merger mania. While a backlash from pinched hospitals has cut into revenue, temp firms see chances for growth with spiraling nurse shortage. AB - While hospitals may be looking for savings by cutting back on temporary personnel, staffing agencies still see potential for growth. That's one of the factors fueling the mergers that have been sweeping the field in recent months as agencies look to diversify and grow. "Big players are looking to get bigger," says analyst Barry Asin, left, a chief analyst at Staffing Industry Analysts. PMID- 16274182 TI - Focus stays on charity care. New class action filed, as one system wins in court. PMID- 16274183 TI - Hospital CEOs lead the way. They top not-for-profit pay list and expect even more. PMID- 16274184 TI - Saved by the budget. States' revenue gains stanch some Medicaid cuts: study. PMID- 16274185 TI - Asset transfers go unseen: GAO. Dems use report to critique commission proposals. PMID- 16274186 TI - Brailer's IT plans draw fire. Privacy, RHIO concerns surface as contracts awarded. PMID- 16274187 TI - ICU effort saved lives, money: organizers. More than 70 hospitals took part in the Keystone: ICU program. PMID- 16274188 TI - Two Providences seek merger. Deal would create Pacific Northwest powerhouse. PMID- 16274189 TI - In deep. The federal spending spree is squeezing out funds for healthcare. PMID- 16274190 TI - Legal implications. National standards needed to ensure interoperability, prevent fraud in EHRs. PMID- 16274191 TI - Destination RHIO. As regional data networks continue to grow in number, some find financial strength. PMID- 16274192 TI - Evolving and expanding. Wash. county's only hospital anchors network . PMID- 16274193 TI - Corn, soybeans and fiber optics. Ind. county providers make sure network takes root. PMID- 16274194 TI - IT's not just for hospitals. Long-term-care facilities have distinct needs for information systems. PMID- 16274195 TI - Digital medicine and the managed care medical director: a look into the future. PMID- 16274196 TI - A view of preserved systolic function and the approach to the heart failure patient. PMID- 16274197 TI - Multiple benefits of bariatric surgery. PMID- 16274198 TI - Evolving perspectives on disease management. PMID- 16274199 TI - Better outcomes through health and productivity management. PMID- 16274200 TI - Quality of care: where do we go from here? PMID- 16274202 TI - U.S. must adopt ICD-10-CM and ICD-10-PCS. PMID- 16274201 TI - Testimony summer: AHIMA testifies on key HIM issues in three government hearings. PMID- 16274203 TI - E-HIM: from vision to reality. PMID- 16274204 TI - Short-term forecast: experts speak up on ONC's RFI, RFPs, and the year ahead. PMID- 16274205 TI - Partnering for quality healthcare. PMID- 16274206 TI - Physician order entry goes online. PMID- 16274207 TI - Spoliation of medical evidence. PMID- 16274208 TI - Privacy compliance--maintaining the gains. PMID- 16274209 TI - Subacute care units. PMID- 16274210 TI - Countdown to HIPAA claims attachments. PMID- 16274211 TI - Educating the public about personal health records. PMID- 16274212 TI - The legal process and electronic health records. PMID- 16274213 TI - Coding complex bariatric surgery. PMID- 16274214 TI - Coding ethically. PMID- 16274215 TI - Treating physician practices with a 5 percent solution. PMID- 16274216 TI - Rhodopsin activation follows precoupling with transducin: inferences from computational analysis. AB - The electrostatic and shape complementarities between the crystal structures of dark rhodopsin and heterotrimeric transducin (Gt) have been evaluated by exhaustively sampling the roto-translational space of one protein with respect to the other. Structural complementarity, reliability, and consistency with in vitro evidence all converge in the same rhodopsin-Gt complex, showing that the functionally important R135 of the E/DRY motif is almost accessible to the C terminus of Gt(alpha) already in the dark state. The main inference from this study is that activation of rhodopsin and Gt may be concurrent processes, consisting of conformational changes in a supramolecular complex formed prior to the light-induced activation of the photoreceptor. PMID- 16274217 TI - Out-of-shape DNA minor groove binders: induced fit interactions of heterocyclic dications with the DNA minor groove. AB - DB921 and DB911 are benzimidazole-biphenyl isomers with terminal charged amidines. DB911 has a central meta-substituted phenyl that gives it a shape similar to those of known minor groove binding compounds. DB921 has a central para-substituted phenyl with a linear conformation that lacks the appropriate radius of curvature to match the groove shape. It is thus expected that DB911, but not DB921, should be an effective minor groove binder, but we find that DB921 not only binds in the groove but also has an unusually high binding constant in SPR experiments (2.9 x 10(8) M(-)(1), vs 2.1 x 10(7) M(-)(1) for DB911). ITC thermodynamic analysis with an AATT sequence shows that the stronger binding of DB921 is due to a more favorable binding enthalpy relative to that of DB911. CD results support minor groove binding for both compounds but do not provide an explanation for the binding of DB921. X-ray crystallographic analysis of DB921 bound to AATT shows that an induced fit structural change in DB921 reduces the twist of the biphenyl to complement the groove, and places the functional groups in position to interact with bases at the floor of the groove. The phenylamidine of DB921 forms indirect contacts with the bases through a bound water. The DB921 water pair forms a curved binding module that matches the shape of the minor groove and provides a number of strong interactions that are not possible with DB911. This result suggests that traditional views of compound curvature required for minor groove complex formation should be reevaluated. PMID- 16274218 TI - Picosecond structural dynamics of myoglobin following photodissociation of carbon monoxide as revealed by ultraviolet time-resolved resonance Raman spectroscopy. AB - Picosecond protein dynamics of myoglobin in response to structural changes in heme upon CO dissociation were observed in a site-specific fashion for the first time using time-resolved UV resonance Raman spectroscopy. Transient UV resonance Raman spectra showed several phases of intensity changes in both tryptophan and tyrosine Raman bands. Five picoseconds after dissociation, the W18, W16, and W3 bands of tryptophan residues and the Y8a band of tyrosine residues decreased in intensity, followed by recovery of the Y8a band intensity in hundreds of picoseconds and recovery of the tryptophan bands in nanoseconds. These spectral changes suggest that the change in heme structure impulsively drives concerted movement of the EF helical section and that rearrangements toward a deoxy structure occur in the heme vicinity and in the A helix within a time frame of sub-nanoseconds to nanoseconds. PMID- 16274219 TI - P61A mutation in the factor for inversion stimulation results in a thermostable dimeric intermediate. AB - The factor for inversion stimulation (FIS) is a homodimeric DNA-binding protein found in enteric bacteria. FIS consists of 98 residues and self-assembles into an entwined dimer containing a flexible and mostly disordered N-terminus followed by four alpha-helices. Proline 61, which is 100% conserved in FIS homologues, is located at the center of helix B, and its substitution for alanine (P61A) was previously shown to result in nonuniform stabilization of the protein, leading to the appearance of a marginally populated dimeric intermediate in urea denaturation equilibrium studies. Here we show that, in contrast to WT FIS, the thermal denaturation of P61A FIS was incomplete and yielded a transition curve that was independent of FIS concentration, suggesting the presence of a dimeric intermediate at 90 degrees C. In the presence of urea, the thermal denaturation of P61A FIS became concentration dependent, consistent with the denaturation of the dimeric intermediate. The existence of a thermostable dimeric intermediate of P61A FIS was further confirmed by glutaraldehyde cross-linking experiments at 95 degrees C. Urea denaturation experiments at 90 degrees C revealed a cooperative transition, indicating that the dimeric intermediate of P61A FIS has a solvent protected hydrophobic core. P61A FIS, unlike the WT protein, was found to be resistant to denaturation by low pH, but its thermal denaturation at pH 3.5 revealed a biphasic transition, providing clues about the structure of the dimeric intermediate. From a functional perspective, it is plausible that the full conservation of proline 61 in FIS may serve to limit the stability and proteolytic resistance of this highly regulated transcription factor. PMID- 16274220 TI - Recombinant expression and functional characterization of human hephaestin: a multicopper oxidase with ferroxidase activity. AB - Human hephaestin (Hp) is a transmembrane protein that has been implicated in duodenal iron export. Mutations in the murine hephaestin gene (sla) produce microcytic, hypochromic anemia that is refractory to oral iron therapy. Hp shares approximately 50% sequence identity with the plasma multicopper ferroxidase ceruloplasmin including conservation of residues involved in disulfide bond formation and metal coordination. On the basis of this similarity to ceruloplasmin, human hephaestin may also bind copper and possess ferroxidase activity. To test this hypothesis, human hephaestin cDNA has been cloned by reverse transcription of human duodenal mRNA. Following in vitro mutagenesis to make the encoded polypeptide suitable for expression and purification, the hephaestin cDNA was cloned into the expression vector pNUT and introduced into baby hamster kidney cells. After selection with methotrexate, the baby hamster kidney cells secreted the recombinant human hephaestin into the medium at a level of approximately 2 mg/L. Purification was achieved by a single immunoaffinity chromatography step. As judged by SDS-PAGE, N-terminal sequence analysis, and matrix-assisted laser desorption ionization-time-of-flight mass spectrometry, the purified hephaestin was homogeneous with a mass of 129600 Da, suggesting a carbohydrate content of 7.7%. Inductively coupled plasma mass spectrometry revealed that recombinant hephaestin contained an average of 3.13 atoms of copper per protein molecule. A visible absorption maximum was observed at 607 nm, consistent with the presence of a Type 1 copper site. By using ferrous ammonium sulfate as a substrate, recombinant hephaestin was shown to have ferroxidase activity with a K(m) of 2.1 microM for Fe(II). Lastly, urea PAGE showed that hephaestin was able to catalyze formation of diferric transferrin from Fe(II) and apotransferrin. PMID- 16274221 TI - Subunits of the yeast mitochondrial ADP/ATP carrier: cooperation within the dimer. AB - The mitochondrial ADP/ATP carrier, or Ancp, is a member of the mitochondrial carrier family (MCF). It exchanges ADP and ATP between matrix and intermembrane space. It is postulated from numerous experiments that the inactive Ancp bound to one of its inhibitors (CATR or BA) is a dimer, and it is inferred that the active unit is a dimer, too. However, the structure of beef Ancp bound to CATR obtained at high resolution is that of a monomer. To ascertain the dimeric organization of Ancp, we have constructed covalent tandem dimers of which one "subunit" (protomer) is the wild type and the other is inactive for ADP/ATP exchange. We have chosen either the op1 mutant or another member of the MCF, the phosphate carrier (Picp). Activities of the chimeras were first evaluated in vivo. The Ancp/op1 constructs exchange the adenine nucleotides. The Anc/Pic chimeras are considered as bifunctional forms since they exchange ADP and ATP and transport P(i) within the same cells. We have then controlled the fact that the chimeras are stable in vivo and in vitro. Proteinase K digestion showed that both protomers of Ancp/op1 have similar organization in the membrane. Analyses of kinetic properties indicated that protomers of Ancp/op1 chimeras crosstalk during the nucleotide exchange unlike those of Anc/Pic. However, full inhibition of phosphate uptake by CATR, a very specific inhibitor of Ancp, strongly suggests that the native functional unit of Ancp, and thus of Picp, is a dimer. PMID- 16274222 TI - Structural basis for the functional differences between type I and type II human methionine aminopeptidases. AB - Determination of the crystal structure of human MetAP1 makes it possible, for the first time, to compare the structures of a Type I and a Type II methionine aminopeptidase (MetAP) from the same organism. Comparison of the Type I enzyme with the previously reported complex of ovalicin with Type II MetAP shows that the active site of the former is reduced in size and would incur steric clashes with the bound inhibitor. This explains why ovalicin and related anti angiogenesis inhibitors target Type II human MetAP but not Type I. The differences in both size and shape of the active sites between MetAP1 and MetAP2 also help to explain their different substrate specificity. In the presence of excess Co(2+), a third cobalt ion binds in the active site region, explaining why metal ions in excess can be inhibitory. Also, the N-terminal region of the protein contains three distinct Pro-x-x-Pro motifs, supporting the prior suggestion that this region of the protein may participate in binding to the ribosome. PMID- 16274223 TI - The role of electrostatics in the interaction of the inhibitory region of troponin I with troponin C. AB - We have addressed the electrostatic interactions occurring between the inhibitory region of cardiac troponin I with the C-lobe of troponin C using scanning glycine mutagenesis of the inhibitory region. We report variations in the electric potentials due to mutation of charged residues within this complex based upon the solved NMR structure (1OZS). These results demonstrate the importance of electrostatics within this complex, and it is proposed that electrostatic interactions are integral to the formation and function of larger ternary troponin complexes. To address this hypothesis, we report (15)N NMR relaxation measurements, which suggest that, within a ternary complex involving the C-lobe and the N-terminal region of troponin I (residues 34-71), the inhibitory region maintains the electrostatic interactions with the E-helix of the C-lobe as observed within the binary complex. These results imply that, in solution, the cardiac troponin complex behaves in a manner consistent with that of the crystal structure of the skeletal isoform (1YTZ). A cardiac troponin complex possessing domain orientations similar to that of the skeletal isoform provides structural insights into altered troponin I activities as observed for the familial hypertrophic cardiomyopathy mutation R144G and phosphorylation of Thr142. PMID- 16274224 TI - Structure of PITPbeta in complex with phosphatidylcholine: comparison of structure and lipid transfer to other PITP isoforms. AB - Phosphatidylinositol transfer protein (PITP) is a ubiquitous eukaryotic protein that preferentially binds either phosphatidylinositol or phosphatidylcholine and catalyzes the exchange of these lipids between membranes. Mammalian cytosolic PITPs include the ubiquitously expressed PITPalpha and PITPbeta isoforms (269-270 residues). The crystal structure of rat PITPbeta complexed to dioleoylphosphatidylcholine was determined to 2.18 A resolution with molecular replacement using rat PITPalpha (77% sequence identify) as the phasing model. A structure comparison of the alpha and beta isoforms reveals minimal differences in protein conformation, differences in acyl conformation in the two isoforms, and remarkable conservation of solvent structure around the bound lipid. A comparison of transfer activity by human and rat PITPs, using small unilamellar vesicles with carefully controlled phospholipid composition, indicates that the beta isoforms have minimal differences in transfer preference between PtdIns and PtdCho when donor vesicles contain predominantly PtdCho. When PtdCho and PtdIns are present in equivalent concentrations in donor vesicles, PtdIns transfer occurs at approximately 3-fold the rate of PtdCho. The rat PITPbeta isoform clearly has the most diminished transfer rate of the four proteins studied. With the two rat isoforms, site-directed mutations of two locations within the lipid binding cavity that possess differing biochemical properties were characterized: I84alpha/F83beta and F225alpha/L224beta. The 225/224 locus is more critical in determining substrate specificity. Following the mutation of this locus to the other amino acid, the PtdCho transfer specific activity became PITPalpha (F225L) approximately PITPbeta and PITPbeta (L224F) approximately PITPalpha. The 225alpha/224beta locus plays a modest role in the specificity of both isoforms toward CerPCho. PMID- 16274225 TI - Oxidation of the non-heme iron complex in photosystem II. AB - In photosystem II (PSII), the redox properties of the non-heme iron complex (Fe complex) are sensitive to the redox state of quinones (Q(A/)(B)), which may relate to the electron/proton transfer. We calculated the redox potentials for one-electron oxidation of the Fe complex in PSII [E(m)(Fe)] based on the reference value E(m)(Fe) = +400 mV at pH 7 in the Q(A)(0)Q(B)(0) state, considering the protein environment in atomic detail and the associated changes in protonation pattern. Our model yields the pH dependence of E(m)(Fe) with -60 mV/pH as observed in experimental redox titration. We observed significant deprotonation at D1-Glu244 in the hydrophilic loop region upon Fe complex oxidation. The calculated pK(a) value for D1-Glu244 depends on the Fe complex redox state, yielding a pK(a) of 7.5 and 5.5 for Fe(2+) and Fe(3+), respectively. To account for the pH dependence of E(m)(Fe), a model involving not only D1 Glu244 but also the other titratable residues (five Glu in the D-de loops and six basic residues near the Fe complex) seems to be needed, implying the existence of a network of residues serving as an internal proton reservoir. Reduction of Q(A/B) yields +302 mV and +268 mV for E(m)(Fe) in the Q(A)(-)Q(B)(0) and Q(A)(0)Q(B)(-) states, respectively. Upon formation of the Q(A)(0)Q(B)(-) state, D1-His252 becomes protonated. Forming Fe(3+)Q(B)H(2) by a proton-coupled electron transfer process from the initial state Fe(2+)Q(B)(-) results in deprotonation of D1-His252. The two EPR signals observed at g = 1.82 and g = 1.9 in the Fe(2+)Q(A)(-) state of PSII may be attributed to D1-His252 with variable and fixed protonation, respectively. PMID- 16274226 TI - Thrombomodulin tightens the thrombin active site loops to promote protein C activation. AB - Thrombomodulin (TM) forms a 1:1 complex with thrombin. Whereas thrombin alone cleaves fibrinogen to make the fibrin clot, the thrombin-TM complex cleaves protein C to initiate the anticoagulant pathway. Crystallographic investigations of the complex between thrombin and TMEGF456 did not show any changes in the thrombin active site. Therefore, research has focused recently on how TM may provide a docking site for the protein C substrate. Previous work, however, showed that when the thrombin active site was occupied with substrate analogues labeled with fluorophores, the fluorophores responded differently to active (TMEGF1-6) versus inactive (TMEGF56) fragments of TM. To investigate this further, we have carried out amide H/(2)H exchange experiments on thrombin in the presence of active (TMEGF45) and inactive (TMEGF56) fragments of TM. Both on exchange and off-exchange experiments show changes in the thrombin active site loops, some of which are observed only when the active TM fragment is bound. These results are consistent with the previously observed fluorescence changes and point to a mechanism by which TM changes the thrombin substrate specificity in favor of protein C cleavage. PMID- 16274227 TI - Cofactor activation and substrate binding in pyruvate decarboxylase. Insights into the reaction mechanism from molecular dynamics simulations. AB - We have performed long-term molecular dynamics simulations of pyruvate decarboxylase from Zymomonas mobilis. Nine structures were modeled to investigate mechanistic questions related to binding of the cofactor, thiamin diphosphate (ThDP), and the substrate in the active site. The simulations reveal that the proposed three ThDP-tautomers all can bind in the active site and indicate that the equilibrium is shifted toward 4'-aminopyrimidine ThDP in the absence of substrate. 4'-Aminopyrimidinium ThDP is found to be a likely intermediate in the equilibrium. Mutations of important active site residues, Glu473Ala and Glu50Ala, were modeled to further elucidate their catalytic role. Formation of the catalytic important ylide by deprotonation of ThDP(C2) is investigated. Only the less favored tautomer, 1',4'-iminopyrimidine ThDP (imino-ThDP), could be deprotonated. The two other tautomers of ThDP could not be activated at the C2 position, thus, explaining the mechanistic importance of the less stable imino ThDP. Finally, binding of pyruvate in the active site with the cofactor modeled as the nucleophilic ylide (ylide-ThDP) is studied. The carbonyl group of the substrate forms a hydrogen bond to Tyr290(OH). No hydrogen bond could be identified between ThDP(N4') and the substrate. The geometry of the substrate binding is well-suited for a nucleophilic attack by ylide-ThDP(C2). We propose that a proton relay from His113 via Asp27 and Tyr290 to the carbonyl oxygen atom of the substrate may be involved in the mechanism. PMID- 16274228 TI - Properties of p-hydroxybenzoate hydroxylase when stabilized in its open conformation. AB - p-Hydroxybenzoate hydroxylase is extensively studied as a model for single component flavoprotein monooxygenases. It catalyzes a reaction in two parts: (1) reduction of the FAD in the enzyme by NADPH in response to binding of p hydroxybenzoate to the enzyme and (2) oxidation of reduced FAD with oxygen in an environment free from solvent to form a hydroperoxide, which then reacts with p hydroxybenzoate to form an oxygenated product. These different reactions are coordinated through conformational rearrangements of the protein and the isoalloxazine ring during catalysis. Until recently, it has not been clear how p hydroxybenzoate gains access to the buried active site. In 2002, a structure of a mutant form of the enzyme without substrate was published that showed an open conformation with solvent access to the active site [Wang, J., Ortiz-Maldonado, M., Entsch, B., Massey, V., Ballou, D., and Gatti, D. L. (2002) Proc. Natl. Acad. Sci. U.S.A. 99, 608-613]. The wild-type enzyme does not form high-resolution crystals without substrate. We hypothesized that the wild-type enzyme without substrate also forms an open conformation for binding p-hydroxybenzoate, but only transiently. To test this idea, we have studied the properties of two different mutant forms of the enzyme that are stabilized in the open conformation. These mutant enzymes bind p-hydroxybenzoate very fast, but with very low affinity, as expected from the open structure. The mutant enzymes are extremely inactive, but are capable of slowly forming small amounts of product by the normal catalytic pathway. The lack of activity results from the failure of the mutants to readily form the out conformation required for flavin reduction by NADPH. The mutants form a large fraction of an abnormal conformation of the reduced enzyme with p hydroxybenzoate bound. This conformation of the enzyme is unreactive with oxygen. We conclude that transient formation of this open conformation is the mechanism for sequestering p-hydroxybenzoate to initiate catalysis. This overall study emphasizes the role that protein dynamics can play in enzymatic catalysis. PMID- 16274229 TI - Crystal structures of the wild-type, P1A mutant, and inactivated malonate semialdehyde decarboxylase: a structural basis for the decarboxylase and hydratase activities. AB - Malonate semialdehyde decarboxylase (MSAD) from Pseudomonas pavonaceae 170 is a tautomerase superfamily member that converts malonate semialdehyde to acetaldehyde by a mechanism utilizing Pro-1 and Arg-75. Pro-1 and Arg-75 have also been implicated in the hydratase activity of MSAD in which 2-oxo-3 pentynoate is processed to acetopyruvate. Crystal structures of MSAD (1.8 A resolution), the P1A mutant of MSAD (2.7 A resolution), and MSAD inactivated by 3 chloropropiolate (1.6 A resolution), a mechanism-based inhibitor activated by the hydratase activity of MSAD, have been determined. A comparison of the P1A-MSAD and MSAD structures reveals little geometric alteration, indicating that Pro-1 plays an important catalytic role but not a critical structural role. The structures of wild-type MSAD and MSAD covalently modified at Pro-1 by 3 oxopropanoate, the adduct resulting from the incubation of MSAD and 3 chloropropiolate, implicate Asp-37 as the residue that activates a water molecule for attack at C-3 of 3-chloropropiolate to initiate a Michael addition of water. The interactions of Arg-73 and Arg-75 with the C-1 carboxylate group of the adduct suggest these residues polarize the alpha,beta-unsaturated acid and facilitate the addition of water. On the basis of these structures, a mechanism for the inactivation of MSAD by 3-chloropropiolate can be formulated along with mechanisms for the decarboxylase and hydratase activities. The results also provide additional evidence supporting the hypothesis that MSAD and trans-3 chloroacrylic acid dehalogenase, a tautomerase superfamily member preceding MSAD in the trans-1,3-dichloropropene degradation pathway, diverged from a common ancestor but retained the key elements for the conjugate addition of water. PMID- 16274230 TI - Role of the tetrahemic subunit in Desulfovibrio vulgaris hildenborough formate dehydrogenase. AB - In the anaerobic sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough (DvH), the genome sequencing revealed the presence of three operons encoding formate dehydrogenases. fdh1 encodes an alphabetagamma trimeric enzyme containing 11 heme binding sites; fdh2 corresponds to an alphabetagamma trimeric enzyme with a tetrahemic subunit; fdh3 encodes an alphabeta dimeric enzyme. In the present work, spectroscopic measurements demonstrated that the reduction of cytochrome c(553) was obtained in the presence of the trimeric FDH2 and not with the dimeric FDH3, suggesting that the tetrahemic subunit (FDH2C) is essential for the interaction with this physiological electron transfer partner. To further study the role of the tetrahemic subunit, the fdh2C gene was cloned and expressed in Desulfovibrio desulfuricans G201. The recombinant FDH2C was purified and characterized by optical and NMR spectroscopies. The heme redox potentials measured by electrochemistry were found to be identical in the whole enzyme and in the recombinant subunit, indicating a correct folding of the recombinant protein. The mapping of the interacting site by 2D heteronuclear NMR demonstrated a similar interaction of cytochrome c(553) with the native enzyme and the recombinant subunit. The presence of hemes c in the gamma subunit of formate dehydrogenases is specific of these anaerobic sulfate-reducing bacteria and replaces heme b subunit generally found in the enzymes involved in anaerobic metabolisms. PMID- 16274231 TI - Solution structure of the C-terminal transcriptional activator domain of FixJ from Sinorhizobium meliloti and its recognition of the fixK promoter. AB - FixJ is a response regulator of the two-component signal transduction pathway involved in the transcriptional activation of nitrogen fixation genes of Sinorhizobium meliloti. Upon phosphorylation, FixJ transcriptionally activates the fixK and nifA promoters. We identified a FixJ recognition sequence of 16 bp in the high affinity binding site of the fixK promoter by means of a gel shift assay. In addition, the solution structure of the truncated C-terminal DNA binding domain of FixJ (FixJC) was solved by NMR spectroscopy. FixJC contains five alpha-helices that encode a typical helix-turn-helix motif as a potential DNA binding core with the highest structural similarity toward the C-terminal DNA binding domain of NarL. The addition of the DNA fragment containing the recognition sequence of the high affinity FixJ binding site resulted in intermediate to slow exchange interactions on the NMR time scale in the spectrum of FixJC, while the exchange was rapid in the case of control DNA. These spectral data suggest that more than one molecule of FixJC binds to the recognition sequence, although FixJC alone is present in monomeric form in solution. This result is consistent with a scenario in which a transcriptionally active species of FixJ is a homodimer of the phosphorylated form. PMID- 16274232 TI - Discovery of acetylcholinesterase peripheral anionic site ligands through computational refinement of a directed library. AB - The formation of beta-amyloid plaques in the brain is a key neurodegenerative event in Alzheimer's disease. Small molecules capable of binding to the peripheral anionic site of acetylcholinesterase (AChE) have been shown to inhibit the AChE-induced aggregation of the beta-amyloid peptide. Using the combination of a computational docking model and experimental screening, five compounds that completely blocked the amyloidogenic effect of AChE were rapidly identified from an approximately 200-member library of compounds designed to disrupt protein protein interactions. Critical to this docking model was the inclusion of two explicit water molecules that are tightly bound to the enzyme. Interestingly, none of the tested compounds inhibited the related enzyme butyrylcholinesterase (BuChE) up to their aqueous solubility limits. These compounds are among the most potent inhibitors of amyloid beta-peptide aggregation and are equivalent only to propidium, a well-characterized AChE peripheral anionic site binder and aggregation inhibitor. PMID- 16274233 TI - Interactive domains for chaperone activity in the small heat shock protein, human alphaB crystallin. AB - Protein pin arrays identified seven interactive sequences for chaperone activity in human alphaB crystallin using natural lens proteins, beta(H) crystallin and gammaD crystallin, and in vitro chaperone target proteins, alcohol dehydrogenase and citrate synthase. The N-terminal domain contained two interactive sequences, (9)WIRRPFFPFHSP(20) and (43)SLSPFYLRPPSFLRAP(58). The alpha crystallin core domain contained four interactive sequences, (75)FSVNLDVK(82) (beta3), (113)FISREFHR(120), (131)LTITSSLS(138) (beta8), and (141)GVLTVNGP(148) (beta9). The C-terminal domain contained one interactive sequence, (157)RTIPITRE(164), that included the highly conserved I-X-I/V motif. Two interactive sequences, (73)DRFSVNLDVKHFS(85) and (131)LTITSSLSDGV(141), belonging to the alpha crystallin core domain were synthesized as peptides and assayed for chaperone activity in vitro. Both synthesized peptides inhibited the thermal aggregation of beta(H) crystallin, alcohol dehydrogenase, and citrate synthase in vitro. Five of the seven chaperone sequences identified by the pin arrays overlapped with sequences identified previously as sequences for subunit-subunit interactions in human alphaB crystallin. The results suggested that interactive sequences in human alphaB crystallin have dual roles in subunit-subunit assembly and chaperone activity. PMID- 16274234 TI - 3D model of the Escherichia coli multidrug transporter MdfA reveals an essential membrane-embedded positive charge. AB - MdfA is an Escherichia coli multidrug transporter of the major facilitator superfamily (MFS) of secondary transporters. Although several aspects of multidrug recognition by MdfA have been characterized, better understanding the detailed mechanism of its function requires structural information. Previous studies have modeled the 3D structures of MFS proteins, based on the X-ray structure of LacY and GlpT. However, because of poor sequence homology, between LacY, GlpT, and MdfA additional constraints were required for a reliable homology modeling. Using an algorithm that predicts the angular orientation of each transmembrane helix (TM) (kPROT), we obtained a remarkably similar pattern for the 12 TMs of MdfA and those of GlpT and LacY, suggesting that they all have similar helix packing. Consequently, a 3D model was constructed for MdfA by structural alignment with LacY and GlpT, using the kPROT results as an additional constraint. Further refinement and a preliminary evaluation of the model were achieved by correlated mutation analysis and the available experimental data. Surprisingly, in addition to the previously characterized membrane-embedded glutamate at position 26, the model suggests that Asp34 and Arg112 are located within the membrane, on the same face of the cavity as Glu26. Importantly, Arg112 is evolutionarily conserved in secondary drug transporters, and here we show that a positive charge at this position is absolutely essential for multidrug transport by MdfA. PMID- 16274235 TI - A role for the protein in internal electron transfer to the catalytic center of cytochrome c oxidase. AB - Internal electron transfer (ET) to heme a(3) during anaerobic reduction of oxidized bovine heart cytochrome c oxidase (CcO) was studied under conditions where heme a and Cu(A) were fully reduced by excess hexaamineruthenium. The data show that ET to heme a(3) is controlled by the state of ionization of a single protolytic residue with a pK(a) of 6.5 +/- 0.2. On the basis of the view that ET to the catalytic site is limited by coupled proton transfer, this pK(a) was attributed to Glu60 which is located at the entrance of the proton-conducting K channel on the matrix side of CcO. It is proposed that Glu60 controls proton entry into the channel. However, even with this channel open, there is the second factor that regulates ET, and this is ascribed to the rate of proton diffusion in the channel. In addition, it is concluded that proton transfer in the K channel is reversibly inhibited by the detergent Triton X-100. It is also found that the rate of ET to heme a(3) in the as-isolated resting enzyme and in CcO "activated" by reaction of fully reduced enzyme with O(2) is the same, implying that the catalytic sites of these two forms of oxidized enzyme are essentially identical. PMID- 16274236 TI - Modeling the endosomal escape of cell-penetrating peptides: transmembrane pH gradient driven translocation across phospholipid bilayers. AB - Cell-penetrating peptides (CPPs) are able to mediate the efficient cellular uptake of a wide range of cargoes. Internalization of a number of CPPs requires uptake by endocytosis, initiated by binding to anionic cell surface heparan sulfate (HS), followed by escape from endosomes. To elucidate the endosomal escape mechanism, we have modeled the process for two CPPs: penetratin (pAntp) and the N-terminal signal peptide of the unprocessed bovine prion protein (bPrPp). Large unilamellar phospholipid vesicles (LUVs) were produced encapsulating either peptide, and an ionophore, nigericin, was used to create a transmembrane pH gradient (DeltapH(mem), inside acidic) similar to the one arising in endosomes in vivo. In the absence of DeltapH(mem), no pAntp escape from the LUVs is observed, while a fraction of bPrPp escapes. In the presence of DeltapH(mem), a significant amount of pAntp escapes and an even higher degree of bPrPp escape takes place. These results, together with the differences in kinetics of escape, indicate different escape mechanisms for the two peptides. A minimum threshold peptide concentration exists for the escape of both peptides. Coupling of the peptides to a cargo reduces the fraction escaping, while complexation with HS significantly hinders the escape. Fluorescence correlation spectroscopy results show that during the escape process the LUVs are intact. Taken together, these results suggest a model for endosomal escape of CPPs: DeltapH(mem)-mediated mechanism, following dissociation from HS of the peptides, above a minimum threshold peptide concentration, in a process that does not involve lysis of the vesicles. PMID- 16274237 TI - Dynamic function of the alkyl spacer of acetogenins in their inhibitory action with mitochondrial complex I (NADH-ubiquinone oxidoreductase). AB - Studies on the inhibitory mechanism of acetogenins, the most potent inhibitors of mitochondrial complex I (NADH-ubiquinone oxidoreductase), are useful for elucidating the structural and functional features of the terminal electron transfer step of this enzyme. Previous studies of the structure-activity relationship revealed that except for the alkyl spacer linking the two toxophores (i.e., the hydroxylated THF and the gamma-lactone rings), none of the multiple functional groups of these inhibitors is essential for potent inhibition. To elucidate the function of the alkyl spacer, two sets of systematically selected analogues were synthesized. First, the length of the spacer was varied widely. Second, the local flexibility of the spacer was specifically reduced by introducing multiple bond(s) into different regions of the spacer. The optimal length of the spacer for inhibition was approximately 13 carbon atoms. The decrease in the strength of the inhibitory effect caused by elongating the spacer from 13 carbons was much more drastic than that caused by shortening. Local flexibility in a specific region of the spacer was not important for the inhibition. These observations indicate that the active conformation of the spacer is not an extended form, and is not necessarily restricted to a certain rigid shape. Moreover, an analogue in which a spacer covering 10 carbon atoms was hardened into a rodlike shape still maintained a potent inhibitory effect. Our results strongly suggest that the spacer portion is free from steric congestion arising from the putative binding site probably because there is no cavity-like binding site for the spacer portion. The manner of acetogenin binding to the enzyme may not be explained by a simple "key and keyhole" analogy. PMID- 16274238 TI - Engineering mouse apolipoprotein A-I into a monomeric, active protein useful for structural determination. AB - Apolipoprotein AI (apoAI), the major protein component of HDL, is one of the best predictors of coronary artery disease (CAD), with high apoAI and HDL levels being correlated with low occurrences of CAD. The primary function of apoAI is to recruit phospholipid and cholesterol for assembly of HDL particles. Like other exchangeable apolipoproteins, lipid-free apoAI forms a mixture of different oligomers even at 1.0 mg/mL. This self-association property of the exchangeable apolipoproteins is closely associated with the lipoprotein-binding activity of this protein family. It is unclear if the self-association property of apolipoprotein is required for its lipoprotein-binding activity. We developed a novel method for engineering an oligomeric protein to a monomeric, biologically active protein. Using this method, we generated a monomeric mouse apoAI mutant that is active. This mutant contains the first 216 residues of mouse apoAI and replaces six hydrophobic residues with either polar or smaller hydrophobic residues at the defined positions (V118A/A119S/L121Q/T191S/T195S/T199S). Cross linking results show that this mutant is greater than 90% monomeric at 8 mg/mL. CD, DSC, and NMR results indicate that the mutant maintains an identical secondary, tertiary structure and stability as those of the wild-type mouse apoAI. Lipid-binding assays suggest that the mutant shares an equal lipoprotein binding activity as that of the wild-type apoAI. In addition, both the monomeric mutant and the wild-type protein make nearly identical rHDL particles. With this monomeric mouse apoAI, high-quality NMR data has been collected, allowing for the NMR structural determination of lipid-free apoAI. On the basis of these results, we conclude that this apoAI mutant is a monomeric, active apoAI useful for structural determination. PMID- 16274239 TI - Functional effects of glycosylation at Asn-579 of the epidermal growth factor receptor. AB - We have investigated functional effects of glycosylation at N(579) of the epidermal growth factor receptor (EGFR). Our previous study showed that the population of cell-surface expressed EGFRs in A431 cells, a human epidermoid carcinoma cell line, is composed of two subpopulations that differ by glycosylation at N(579) [Zhen et al. (2003) Biochemistry 42, 5478-5492]. To characterize the subpopulation of receptors not glycosylated at N(579), we established a 32D cell line expressing a point mutant of the EGFR (N579Q), which cannot be glycosylated at this position. Analysis of epitope accessibility suggests that the lack of glycosylation at N(579) weakens auto-inhibitory tether interactions, and cross-linking experiments suggest a somewhat elevated level of preformed N579Q-EGFR dimers in the absence of ligand relative to wild-type EGFR (WT-EGFR). However, ligand drives the majority of N579Q-EGFR dimerization, suggesting that untethering, while necessary, is not sufficient to drive dimerization. Ligand-binding experiments reveal a much greater fraction of N579Q EGFRs in a high-affinity state compared to the fraction of WT-EGFRs in a high affinity state. However, differences in the kinetic association and dissociation rates indicate that the high-affinity states of the WT and the N579Q receptors are distinct. EGF-stimulated phosphorylation in cells expressing N579Q-EGFRs results in notable differences in the pattern of tyrosine phosphorylated proteins compared with that obtained in cells expressing WT-EGFRs. Moreover, although WT EGFRs confer cell survival in 32D cells in the absence of interleukin-3 and EGF, we found that receptors lacking glycosylation at N(579) do not. This is the first study of which we are aware to show that selective glycosylation of a specific N glycosylation site can produce two functionally distinct receptors. PMID- 16274240 TI - Decoding protein-protein interactions through combinatorial chemistry: sequence specificity of SHP-1, SHP-2, and SHIP SH2 domains. AB - A general, combinatorial library method for the rapid identification of high affinity peptide ligands of protein modular domains is reported. The validity of this method has been demonstrated by determining the sequence specificity of four Src homology 2 (SH2) domains derived from protein tyrosine phosphatase SHP-1 and SHP-2 and inositol phosphatase SHIP. A phosphotyrosyl (pY) peptide library was screened against the SH2 domains, and the beads that carry high-affinity ligands of the SH2 domains were identified and peptides were sequenced by partial Edman degradation and mass spectrometry. The results reveal that the N-terminal SH2 domain of SHP-2 is capable of recognizing four different classes of pY peptides. Binding competition studies suggest that the four classes of pY peptides all bind to the same site on the SH2 domain surface. The C-terminal SH2 domains of SHP-1 and SHP-2 and the SHIP SH2 domain each bind to pY peptides of a single consensus sequence. Database searches using the consensus sequences identified most of the known as well as many potential interacting proteins of SHP-1 and/or SHP-2. Several proteins are found to bind to the SH2 domains of SHP-1 and SHP-2 through a new, nonclassical ITIM motif, (V/I/L)XpY(M/L/F)XP, which corresponds to the class IV peptides selected from the pY library. The combinatorial library method should be generally applicable to other protein domains. PMID- 16274241 TI - Perturbation of yeast 3-phosphoglycerate kinase reaction mixtures with ADP: transient kinetics of formation of ATP from bound 1,3-bisphosphoglycerate. AB - 3-Phosphoglycerate kinase (PGK) is the first ATP-producing enzyme in glycolysis: ADP + 1,3-bisphosphoglycerate (bPG) <--> ATP + 3-phosphoglycerate (PG). Whereas extensive studies have been carried out on its structure, there is less information about its reaction pathway, which is usually studied in the reverse direction because of the instability of bPG. We studied the transients of the PGK reaction by chemical sampling in a rapid quench flow apparatus, using [gamma (32)P]ATP, in 30% methanol at 4 degrees C to decrease k(cat). There were two types of experiment, both at low PG concentrations to prevent bPG release. In the first, reaction mixtures were quenched in acid at different times (from 4 ms) and the bPG concentrations were determined. This type gave information about the ATP binding and phospho-transfer steps. In the second, PGK reaction mixtures at equilibrium were perturbed by the injection of ADP, the new mixtures aged for different times and quenched in acid, and the bPG concentrations were determined. This gave information about the kinetics of the binding of ADP to a PGK intermediate. The data from the two types of experiments were fitted to simple schemes and then treated together by a global fitting procedure using a five-step pathway, deduced from previous structural studies. Under our conditions, it appears that (1) a binary PGK.bPG complex is an important intermediate on the reaction pathway, i.e., that ADP is released before bPG, (2) ADP binds to a "closed" conformation in the PGK.bPG complex, and (3) the PGK reaction can be studied in the physiologically important direction without having to handle bPG. PMID- 16274242 TI - Thermodynamics and kinetics of formation of the alkaline state of a Lys 79- >Ala/Lys 73-->His variant of iso-1-cytochrome c. AB - The alkaline transition kinetics of a Lys 73-->His (H73) variant of iso-1 cytochrome c are triggered by three ionizable groups [Martinez, R. E., and Bowler, B. E. (2004) J. Am. Chem. Soc. 126, 6751-6758]. To eliminate ambiguities caused by overlapping phases due to formation of the Lys 79 alkaline conformer and proline isomerization associated with the His 73 alkaline conformer, we mutated Lys 79 to Ala in the H73 variant (A79H73). The stability and guanidineHCl m-values of the A79H73 and H73 variants at pH 7.5 are the same. The Ala 79 mutation causes formation of the alkaline conformer to depend on [NaCl]. The salt dependence saturates at 500 mM NaCl, and the thermodynamics of alkaline state formation for the A79H73 and H73 variants become identical. The salt dependence is consistent with loss of an electrostatic contact between Lys 79 and heme propionate D in the A79H73 variant. The kinetics of alkaline state formation for the A79H73 variant support the three trigger group model developed for the H73 variant, with the primary trigger, pK(HL), being ionization of His 73. The low pH ionization, pK(H1), is perturbed by the Ala 79 mutation indicating that this ionization is modulated by the buried hydrogen bond network involving heme propionate D. The A79H73 variant has a high spin heme above pH 9 suggesting that the high pH ionization, pK(H2), involves a high spin heme conformer. The proline isomerization phase is modulated by both pK(HL) and pK(H2) indicating that it is sensitive to protein conformation. PMID- 16274243 TI - CouO and NovO: C-methyltransferases for tailoring the aminocoumarin scaffold in coumermycin and novobiocin antibiotic biosynthesis. AB - During the biosynthesis of the streptomycete aminocoumarin antibiotics novobiocin and the dimeric coumermycin A(1), the bicyclic coumarin scaffold is C-methylated adjacent to the phenolic oxygen. The SAM-dependent C-methyltransferases NovO and CouO have been heterologously expressed and purified from Escherichia coli and shown to act after the aminocoumarin ring has been constructed by prior action of Nov/CouHIJK. Neither C-methyltransferase works on the tyrosyl-derived S pantetheinyl intermediates tethered to NovH or on the subsequently released free aminocoumarin. NovL ligates the aminocoumarin to prenylhydroxybenzoate to yield novobiocic acid, which is the substrate for NovO before it is O-glycosylated by NovM. In coumermycin assembly, the corresponding ligase CouL makes the bis-amide by tandem ligation of two aminocoumarins to a dicarboxypyrrole. CouO works on both the mono- and bis-amides for mono- and di-C-methylation adjacent to the phenolic hydroxyl before it is glycosylated by CouM. Thus, the specific timing of C-methylation in the aminocoumarin antibiotic pathways is established. PMID- 16274245 TI - Conjoined hemoglobins. Loss of cooperativity and protein-protein interactions. AB - Hemoglobin cross-linked as a bis(isophthalamide) of the epsilon-amino groups of lysine 82 of each beta-subunit binds and releases oxygen with a Hill coefficient indicative of cooperative oxygen binding (typically approximately 2.0). However, connecting two such cross-linked tetramers with a relatively short covalent linkage produces cross-linked bis-tetramers that bind oxygen with Hill coefficients near unity. To separate the effect of the linkages from the effects of protein-protein interactions in the conjoined proteins, reagents (1 and 2) were used to produce bis-tetramers (A and B). These have a considerably greater distance between cross-linked tetramers than earlier examples. Yet, the bis tetramers (A and B) bind oxygen with minimal cooperativity (n(50) = 1.4, 1.2). To assess the effect of the linkage itself, cross-linked tetramers (Cand D) were prepared from reactions with the same reagents. These bind oxygen with cooperativity similar to that of cross-linked tetramers without the extended chain (C, n(50) = 2.0; D, n(50) = 1.8). Other tetramers (E and F) with flexible, saturated hydrocarbon appendages were also prepared. These also showed cooperativity in oxygen binding (E, n(50) = 1.7; F, n(50) = 1.8) despite their high degree of hydrophobicity. Thus, the intertetrameric linkages themselves do not induce the loss of cooperativity, leading to the conclusion that solution effects of the tetramers upon one another are the source of the decline in cooperativity: protein-protein interactions are most significant in disrupting the cooperativity of the bis-tetramers, regardless of the span or composition of the linker. This suggests that effects of oligomerization of hemoglobin within red cell substitutes should be considered in terms of such interactions. PMID- 16274244 TI - Mechanism of allosteric regulation of Dnmt1's processivity. AB - We have analyzed the relationship between the allosteric regulation and processive catalysis of DNA methyltransferase 1 (Dnmt1). Processivity is described quantitatively in terms of turnover rate, DNA dissociation rate, and processivity probability. Our results provide further evidence that the active site and the allosteric sites on Dnmt1 can bind DNA independently. Dnmt1's processive catalysis on unmethylated DNA is partially inhibited when the allosteric site binds unmethylated DNA and fully inhibited when the allosteric site binds a single-stranded oligonucleotide inhibitor. The partial inhibition by unmethylated DNA is caused by a decrease in the turnover rate and an increase in the substrate DNA dissociation rate. Processive catalysis with premethylated DNA is not affected if the allosteric site is exposed to premethylated DNA but is fully inhibited if the allosteric site binds unmethylated DNA or poly(dA-dT). In sum, the occupancy of the allosteric site modulates the enzyme's commitment to catalysis, which reflects the nature of the substrate and the DNA bound at the allosteric site. Our in vitro results are consistent with the possibility that the processive action of Dnmt1 may be regulated in vivo by specific regulatory nucleic acids such as DNA, RNA, or poly(ADP-ribose). PMID- 16274246 TI - Binding of cationic porphyrin to isolated DNA and nucleoprotein complex: quantitative analysis of binding forms under various experimental conditions. AB - We studied the complex formation of tetrakis(4-N-methylpyridyl)porphyrin (TMPyP) with double stranded DNAs and T7 phage nucleoprotein complex. We analyzed the effect of base pair composition of DNA, the presence of capsid protein, and the composition of the microenvironment on the distribution of TMPyP between binding forms as determined by the decomposition of porphyrin absorption spectra. No difference was found in the amount of bound TMPyP between DNAs of various base compositions; however, the ratio of TMPyP binding forms depends on the AT/GC ratio. The presence of protein capsid opposes the binding of TMPyP to DNA. This behavior offers a possibility to investigate the protein capsid integrity due to the analysis of porphyrin binding. Increasing ionic strength of monovalent ions decreases the amount of bound porphyrin through the inhibition of intercalation, but does not influence the quantity of groove-binding forms when TMPyP interacts with isolated DNA. In the case of the nucleoprotein complex the groove-binding is also inhibited already at 140 mM ionic strength. The presence of 1 mM divalent cations (Mg(2+), Ca(2+), Cu(2+) and Ni(2+)) in a buffer solution of 70 mM ionic strength does not influence significantly the free to bound ration of TMPyP when it interacts with isolated DNA. The contribution of binding forms is remarkably different in Mg(2+)/Ca(2+) and Cu(2+)/Ni(2+) containing solutions. Transition metals significantly decrease the binding sites for intercalation in both DNA and nucleoprotein complex, but facilitate the groove-binding of TMPyP to isolated DNA. PMID- 16274247 TI - A trans acting ribozyme that phosphorylates exogenous RNA. AB - The structural complexity required for substrate recognition within an active site constrains the evolution of novel catalytic functions. To evaluate those constraints within populations of incipient ribozymes, we performed a selection for kinase ribozymes under conditions that allowed competition for phosphorylation at nine candidate sites. Two candidate sites are the hydroxyl groups on a "quasi-diffusible" chloramphenicol (Cam) moiety tethered to the evolving library through an inert, flexible linker. A subtractive step was included to allow only seven ribose 2' hydroxyls to compete with the two Cam hydroxyls for phosphorylation. After the library was incubated with gamma-thio ATP (ATPgammaS), active species were recovered from a polyacrylamide gel containing [(N-acryloylamino)phenyl] mercury (APM) and amplified for further cycles of selection. Activity assays on selected isolates and truncated derivatives identified the essential secondary structure of the dominant RNA motif. Phosphorylation was independent of the Cam moiety, indicating ribose 2' phosphorylation. The dominant motif was separated into catalytic "ribozyme" and "substrate" strands. Partial alkaline digestion of the substrate strand before and after phosphorylation identified the precise modification site as the first purine (R) within the required sequence 5'-RAAAANCG-3'. The reaction shows approximately 10-fold preference for ATPgammaS over ATP and is independent of pH over a wide range (5.5-8.9), consistent with a dissociative reaction mechanism that is rate-limited by formation of a metaphosphate transition state. Divalent metal ions are required, with a slight preference of Mn(2+) > Mg(2+) > Ca(2+). Lack of reactivity in [Co(NH(3))(6)](3+) indicates a requirement for inner sphere contact with the metal ion, either for structural stabilization, catalysis, or both. PMID- 16274248 TI - Bipolar tetraether lipids: chain flexibility and membrane polarity gradients from spin-label electron spin resonance. AB - Membranes of thermophilic Archaea are composed of unique tetraether lipids in which C40, saturated, methyl-branched biphytanyl chains are linked at both ends to polar groups. In this paper, membranes composed of bipolar lipids P2 extracted from the acidothermophile archaeon Sulfolobus solfataricus are studied. The biophysical basis for the membrane formation and thermal stability is investigated by using electron spin resonance (ESR) of spin-labeled lipids. Spectral anisotropy and isotropic hyperfine couplings are used to determine the chain flexibility and polarity gradients, respectively. For comparison, similar measurements have been carried out on aqueous dispersions of diacyl reference lipid dipalmitoyl phosphatidylcholine and also of diphytanoyl phosphatidylcholine, which has methyl-branched chains. At a given temperature, the bolaform lipid chains are more ordered and less flexible than in normal bilayer membranes. Only at elevated temperatures (80 degrees C) does the flexibility of the chain environment in tetraether lipid assemblies approach that of fluid bilayer membranes. The height of the hydrophobic barrier formed by a monolayer of archaebacterial lipids is similar to that in conventional fluid bilayer membranes, and the permeability barrier width is comparable to that formed by a bilayer of C16 lipid chains. At a mole ratio of 1:2, the tetraether P2 lipids mix well with dipalmitoyl phosphatidylcholine lipids and stabilize conventional bilayer membranes. The biological as well as the biotechnological relevance of the results is discussed. PMID- 16274249 TI - Concanamycin and indolyl pentadieneamide inhibitors of the vacuolar H+-ATPase bind with high affinity to the purified proteolipid subunit of the membrane domain. AB - The macrolide antibiotic concanamycin is a potent and specific inhibitor of the vacuolar H(+)-ATPase (V-ATPase), binding to the V(0) membrane domain of this eukaryotic acid pump. Although binding is known to involve the 16 kDa proteolipid subunit, contributions from other V(0) subunits are possible that could account for the apparently different inhibitor sensitivities of pump isoforms in vertebrate cells. In this study, we used a fluorescence quenching assay to directly examine the roles of V(0) subunits in inhibitor binding. Pyrene-labeled V(0) domains were affinity purified from Saccharomyces vacuolar membranes, and the 16 kDa proteolipid was subsequently extracted into chloroform and methanol and purified by size exclusion chromatography. Fluorescence from the isolated proteins was strongly quenched by nanomolar concentrations of both concanamycin and an indolyl pentadieneamide compound, indicating high-affinity binding of both natural macrolide and synthetic inhibitors. Competition studies showed that these inhibitors bind to overlapping sites on the proteolipid. Significantly, the 16 kDa proteolipid in isolation was able to bind inhibitors as strongly as V(0) did. In contrast, proteolipids carrying mutations that confer resistance to both inhibitors showed no binding. We conclude that the extracted 16 kDa proteolipid retains sufficient fold to form a high-affinity inhibitor binding site for both natural and synthetic V-ATPase inhibitors and that the proteolipid contains the major proportion of the structural determinants for inhibitor binding. The role of membrane domain subunit a in concanamycin binding and therefore in defining the inhibitor binding properties of tissue-specific V-ATPases is critically re assessed in light of these data. PMID- 16274250 TI - Isomers of epidermal growth factor with Ser --> Cys mutation at the N-terminal sequence: isomerization, stability, unfolding, refolding, and structure. AB - The structure of human epidermal growth factor (EGF, 53 amino acids) comprises three distinct loops (A, B, and C) connected correspondingly by the three native disulfide bonds, Cys(6)-Cys(20), Cys(14)-Cys(31), and Cys(33)-Cys(42). The connection of Cys(6) and Cys(20) forming the N-terminal A loop is essential for the biological activity of EGF [Barnham et al. (1998) Protein Sci. 7, 1738-1749] and has also been shown to represent a major kinetic trap in the oxidative folding of EGF [Chang et al. (2001) J. Biol. Chem. 276, 4845-4852]. To further understand the chemical nature of this kinetic trap, we have prepared three EGF mutants each with a single Ser --> Cys mutation at Ser residues (Ser(2), Ser(4), and Ser(9)) flanking Cys(6). This allows competition between Cys(6) and mutated Cys(2), Cys(4), and Cys(9) to link with Cys(20) and to form EGF isomers containing different sizes of the A loop. The results show that, in the cases of EGF(S2C) and EGF(S4C), native Cys(6)-Cys(20) is favored over Cys(2)-Cys(20) and Cys(4)-Cys(20) by 4.5- and 9-fold, respectively, in the state of equilibrium. However, in the case of EGF(S9C), a non-native Cys(9)-Cys(20) is thermodynamically more stable than the native Cys(6)-Cys(20) by a free-energy difference (DeltaG degrees ) of 1.12 kcal/mol. Implications of these data in the formation of kinetic trap of EGF folding are discussed. Stabilized isomers of EGF were further generated from denaturation of wild-type and mutant EGF via the method of disulfide scrambling. Properties of these diverse isomers of EGF, including their isomerization, stability, unfolding, refolding, and disulfide structures, are described in this paper. PMID- 16274251 TI - Insights into human Lck SH3 domain binding specificity: different binding modes of artificial and native ligands. AB - We analyzed the ligand binding specificity of the lymphocyte specific kinase (Lck) SH3 domain. We identified artificial Lck SH3 ligands using phage display. In addition, we analyzed Lck SH3 binding sites within known natural Lck SH3 binding proteins using an Lck specific binding assay on membrane-immobilized synthetic peptides. On one hand, from the phage-selected peptides, representing mostly special class I' ligands, a well-defined consensus sequence was obtained. Interestingly, a histidine outside the central polyproline motif contributes significantly to Lck SH3 binding affinity and specificity. On the other hand, we confirmed previously mapped Lck SH3 binding sites in ADAM15, HS1, SLP76, and NS5A, and identified putative Lck SH3 binding sites of Sam68, FasL, c-Cbl, and Cbl-b. Without exception, the comparatively diverse Lck SH3 binding sites of all analyzed natural Lck SH3 binding proteins emerged as class II proteins. Possible explanations for the observed variations between artificial and native ligands which are not due to significant K(D) value differences as shown by calculating Lck SH3 affinities of artificial peptide PD1-Y(-3)R as well as for peptides comprising putative Lck SH3 binding sites of NS5A, Sos, and Sam68-are discussed. Our data suggest that phage display, a popular tool for determining SH3 binding specificity, must-at least in the case of Lck-not irrevocably mirror physiologically relevant protein-ligand interactions. PMID- 16274252 TI - Tubulin assembly, taxoid site binding, and cellular effects of the microtubule stabilizing agent dictyostatin. AB - (-)-Dictyostatin is a sponge-derived, 22-member macrolactone natural product shown to cause cells to accumulate in the G2/M phase of the cell cycle, with changes in intracellular microtubules analogous to those observed with paclitaxel treatment. Dictyostatin also induces assembly of purified tubulin more rapidly than does paclitaxel, and nearly as vigorously as does dictyostatin's close structural congener, (+)-discodermolide (Isbrucker et al. (2003), Biochem. Pharmacol. 65, 75-82). We used synthetic (-)-dictyostatin to study its biochemical and cytological activities in greater detail. The antiproliferative activity of dictyostatin did not differ greatly from that of paclitaxel or discodermolide. Like discodermolide, dictyostatin retained antiproliferative activity against human ovarian carcinoma cells resistant to paclitaxel due to beta-tubulin mutations and caused conversion of cellular soluble tubulin pools to microtubules. Detailed comparison of the abilities of dictyostatin and discodermolide to induce tubulin assembly demonstrated that the compounds had similar potencies. Dictyostatin inhibited the binding of radiolabeled discodermolide to microtubules more potently than any other compound examined, and dictyostatin and discodermolide had equivalent activity as inhibitors of the binding of both radiolabeled epothilone B and paclitaxel to microtubules. These results are consistent with the idea that the macrocyclic structure of dictyostatin represents the template for the bioactive conformation of discodermolide. PMID- 16274253 TI - Differential regulation of vascular endothelial growth factor receptors (VEGFR) revealed by RNA interference: interactions of VEGFR-1 and VEGFR-2 in endothelial cell signaling. AB - Vascular endothelial growth factor (VEGF) plays a central role in vascular homeostasis. VEGF receptors (VEGFRs) include several subtypes that may have a differential role in endothelial signal transduction, but interactions among these receptors are incompletely understood. In these studies, we designed small interfering RNA (siRNA) duplexes that targeted specific VEGFR subtypes in bovine aortic endothelial cells (BAEC). siRNA-mediated downregulation of VEGFR-2 by its cognate siRNA resulted in a significant attenuation of VEGF-mediated signaling. Compared to control siRNA-treated cells, VEGFR-2 siRNA markedly inhibited VEGF mediated activation of PI3K/Akt/GSK3-beta as well as MAP kinase and PKC pathways. VEGFR-2 siRNA also blocked VEGF-stimulated phosphorylation and dephosphorylation of endothelial nitric oxide synthase (eNOS) at Ser(1179) and Ser(116), respectively. VEGFR-2-specific siRNA had no effect on the abundance of VEGFR-1 protein. By contrast, VEGFR-1-specific siRNA markedly not only downregulated the abundance of VEGFR-1 but also significantly reduced VEGFR-2 protein and mRNA abundance. VEGFR-1 siRNA had no effect on the stability of VEGFR-2 protein or mRNA. However, VEGFR-1 siRNA significantly inhibited VEGFR-2 promoter activity, as determined in luciferase assays using VEGFR-2 promoter fusion constructs in transfected BAEC. Deletion of either the 5' E box or the 3' E box and the GATA element in the VEGFR-2 promoter completely abolished the inhibition of VEGFR-2 promoter activity elicited by VEGFR-1 siRNA. Taken together, our data suggest that VEGFR-1 receptor is a critical determinant of VEGFR-2 abundance, while VEGFR 2 is the key receptor directly responsible for endothelial cell signaling stimulated by VEGF. PMID- 16274257 TI - Antidepressant drug-drug interaction profile update. AB - Drug-drug interactions continue to be underappreciated and misunderstood by most clinicians. Although life-threatening drug interactions are rare, serious clinical consequences, including altered drug response, poor tolerability with reduced medication adherence, and increased costs for care tied to the increased complexity of therapy, are fairly commonplace. Drug interactions may be further complicated by genetic differences in metabolic capacity. Patients who routinely require long-term treatment for depression have an increased likelihood of experiencing a drug-drug interaction since they will take over-the-counter and prescription medications for intercurrent and/or co-morbid illness. Antidepressants can be the object of drug interactions when their metabolic pathways are affected by other substances, or they can precipitate interactions by inhibiting enzyme pathways. Clinicians can improve the short- and long-term outcomes of patients with a depressive disorder by considering the possibility of drug-drug interactions both before prescribing a specific antidepressant and while monitoring for response, adverse effects and patient compliance. PMID- 16274254 TI - Calmodulin regulation and identification of calmodulin binding region of type-3 ryanodine receptor calcium release channel. AB - Ryanodine receptors (RyRs) are a family of intracellular Ca(2+) channels that are regulated by calmodulin (CaM). At low Ca(2+) concentrations (<1 microM), CaM activates RyR1 and RyR3 and inhibits RyR2. At elevated Ca(2+) concentrations (>1 microM), CaM inhibits all three RyR isoforms. Here we report that the regulation of recombinant RyR3 by CaM is sensitive to redox regulation. RyR3 in the presence of reduced glutathione binds CaM with 10-15-fold higher affinity, at low and high Ca(2+) concentrations, compared to in the presence of oxidized glutathione. However, compared to RyR1 assayed at low Ca(2+) concentrations under both reducing and oxidizing conditions, CaM binds RyR3 with reduced affinity but activates RyR3 to a greater extent. Under reducing conditions, RyR1 and RyR3 activities are inhibited with a similar affinity at [Ca(2+)] > 1 microM. Mutagenesis studies demonstrate that RyR3 contains a single conserved CaM binding site. Corresponding amino acid substitutions in the CaM binding site differentially affect CaM binding and CaM regulation of RyR3 and those of the two other isoforms. The results support the suggestion that other isoform dependent regions have a major role in the regulation of RyRs by CaM [Yamaguchi et al. (2004) J. Biol. Chem. 279, 36433-36439]. PMID- 16274258 TI - Topical ketoprofen patch. AB - Although oral nonsteroidal anti-inflammatory drugs (NSAIDs) are effective in the treatment of a variety of acute and chronic pain conditions, their use may be associated with serious systemic adverse effects, particularly gastrointestinal disorders. In order to minimise the incidence of systemic events related to such agents, topical NSAIDs have been developed. Topical NSAIDs, applied as gels, creams or sprays, penetrate the skin, subcutaneous fatty tissue and muscle in amounts that are sufficient to exert a therapeutic effect on peripheral and central mechanisms in the absence of high plasma concentrations. Data indicate that topical NSAIDs are effective at relieving pain in a number of acute and chronic pain indications. This review article discusses the pharmacokinetics, efficacy and tolerability of a new formulation of ketoprofen available as a topical patch. The topical patch containing ketoprofen 100mg as the active principle has been developed using a novel delivery system that dispenses therapeutic doses of the drug directly to the site of injury. Pharmacokinetic data indicate that although plasma levels of ketoprofen are higher when the drug is administered as a patch versus a gel, the total systemic bioavailability of ketoprofen 100 mg administered via a patch is no more than 10% of that reported for ketoprofen 100 mg administered orally. Because the patch facilitates ketoprofen delivery over a 24-hour period, the drug remains continually present in the tissue subjacent to the site of application. High tissue but low plasma ketoprofen concentrations mean that while tissue concentrations are high enough to exert a therapeutic effect, plasma concentrations remain low enough to not result in systemic adverse events caused by elevated serum NSAID levels. Phase III clinical trials in patients with non-articular rheumatism and traumatic painful soft tissue injuries showed that the topical ketoprofen patch was significantly more effective than placebo at reducing pain during daily activities and spontaneous pain after 7 days' treatment. Moreover, the topical ketoprofen patch was well tolerated; adverse events were primarily cutaneous in nature and occurred in a similar number of ketoprofen and placebo recipients suggesting that these events were related to the patch itself rather than the active ingredient. The incidence of gastrointestinal adverse events was low (<8% of all patients), and occurred in a similar proportion of patients receiving ketoprofen and placebo. Thus, the topical ketoprofen patch appears to be a simple, effective and safe therapeutic option for the treatment of local painful inflammation. PMID- 16274259 TI - Analysis of lipid peroxidation and electron microscopic survey of maturation stages during human cataractogenesis: pharmacokinetic assay of Can-C N acetylcarnosine prodrug lubricant eye drops for cataract prevention. AB - Morphological and biophysical techniques described in this study have shown that membrane derangement occurs in human cataractous lenses. The data suggest that these disruptions were globules, vacuoles, multilamellar membranes and clusters of highly undulating membranes. Deleterious structural damage of the lens fibre cell plasma membranes serve as the primary light-scattering centres that cause the observed lens opacity. Nuclear cataract, a major cause of loss of lens transparency in the aging human, has been thought to be associated with oxidative damage, particularly at the site of the nuclear plasma membrane. Phospholipid molecules modified by oxygen accumulate in the lipid bilayer, change its geometry and impair lipid-lipid and protein-lipid interactions in lenticular fibre membranes. Lipid peroxidation (LPO) is a causative and pathogenic factor in cataract. Increased concentrations of primary molecular LPO products (diene conjugates, lipid hydroperoxides, oxy-derivatives of phospholipid fatty acids) and end-fluorescent LPO products have been detected in the lipid moieties of aqueous humour samples and human lenses obtained from patients with senile and complicated cataracts as compared with normal donors. In the present study, a rapid and simple high-performance liquid chromatographic (HPLC) assay for determination of imidazole-containing dipeptides in the aqueous humour of the eye was developed. The method was applied to determine the pharmacokinetic parameters and the time-course of N-acetylcarnosine and L-carnosine-related product in the eye, following a single dosage of topical ocular administration of peptide. Utilising data from pharmacokinetic studies and the specific purity of the N acetylcarnosine (NAC) ingredient as a source of the pharmacological principle L carnosine, we have created an ophthalmic time-release prodrug form including the US FDA-approved carboxymethylcellulose lubricant and other essential ingredients (Can-C, private label Nu-Eyes). This formulation increases the intraocular absorption of L-carnosine in the aqueous humour and optimises its specific antioxidant activity in vivo while reducing the toxic effects of lipid peroxides on the crystalline lens. L-carnosine that enters the aqueous humour can accumulate in the lens tissue for a reasonable period of time. The presence of L carnosine in transparent crystalline lenses during normal aging was detected and its concentration in this case was about 25 microM. At different stages of cataract development, the level of L-carnosine drastically decreased, reaching about 5 microM in ripe human cataracts. However, administration of pure L carnosine (1% solution) to the rabbit eye (instillation or subconjunctival injection) does not lead to accumulation of this natural compound in the aqueous humour at the time level over 30 minutes at a concentration exceeding that in placebo-treated matched eyes, and its effective concentration is exhausted more rapidly. Use of NAC prodrug eye drops optimises the clinical effects of L carnosine in the treatment of ophthalmic disorders (such as prevention and reversal of cataracts in human and animal [canine] eyes). The data provided predict a clinical effect with NAC ophthalmic prodrug, and show that the magnitude and duration of this effect are directly related to the bioavailability of L-carnosine released from NAC in the aqueous humour of the anterior eye segment. The ophthalmic NAC drug shows promise in the treatment of a range of ophthalmic disorders that have a component of oxidative stress in their pathogenesis (including cataract, glaucoma, dry eye, vitreous floaters, inflammatory disorders, and corneal, retinal and systemic diseases [such as diabetes mellitus and its ophthalmic complications]). There is a need for further and better collaboration between Innovative Vision Products' cataract control and ophthalmic services, improved education of people affected by cataract, a commitment that N-acetylcarnosine eye drops will be the preferred treatment before orthodox cataract surgery is attempted, and consideration of outcomes and a possible role of the NAC drug cataract treatment as source of referral for orthodox surgical, ophthalmic and optometric services. PMID- 16274260 TI - Preclinical pharmacology of bilastine, a new selective histamine H1 receptor antagonist: receptor selectivity and in vitro antihistaminic activity. AB - OBJECTIVE: This study aimed to establish the receptor selectivity and antihistaminic activity of bilastine, a new selective antihistamine receptor antagonist. DESIGN AND METHODS: In vitro experiments were conducted using a receptor binding screening panel and guinea-pig and rat tissues. Antihistaminic activity was determined using H1 receptor binding studies and in vitro H1 antagonism studies conducted in guinea-pig tissues and human cell lines. Receptor selectivity was established using a receptor binding screening panel and a receptor antagonism screening conducted in guinea-pig, rat and rabbit tissues. Inhibition of inflammatory mediators was determined through the Schultz-Dale reaction in sensitised guinea-pig ileum. RESULTS: Bilastine binds to histamine H1 receptors as indicated by its displacement of [3H]-pyrilamine from H1-receptors expressed in guinea-pig cerebellum and human embryonic kidney (HEK) cell lines. The studies conducted on guinea-pig smooth muscle demonstrated the capability of bilastine to antagonise H1-receptors. Bilastine is selective for histamine H1 receptors as shown in receptor-binding screening conducted to determine the binding capacity of bilastine to 30 different receptors. The specificity of its H1-receptor antagonistic activity was also demonstrated in a series of in vitro experiments conducted on guinea-pig and rat tissues. The results of these studies confirmed the lack of significant antagonism against serotonin, bradykinin, leukotriene D4, calcium, muscarinic M3-receptors, alpha1-adrenoceptors, beta2 adrenoceptors, and H2- and H3-receptors. The results of the in vitro Schultz-Dale reaction demonstrated that bilastine also has anti-inflammatory activity. CONCLUSIONS: These preclinical studies provide evidence that bilastine has H1- antihistamine activity, with high specificity for H1-receptors, and poor or no affinity for other receptors. Bilastine has also been shown to have anti inflammatory properties. PMID- 16274261 TI - Influence of hepatic impairment on the pharmacokinetics and pharmacodynamics of landiolol hydrochloride, an ultra-short-acting beta1-blocker. AB - OBJECTIVE: To investigate the effects of mild to moderate hepatic impairment on the pharmacokinetics and pharmacodynamics of landiolol hydrochloride, a new ultra short-acting beta1-adrenergic antagonist. METHODS: Six patients with hepatic impairment and six healthy volunteers were enrolled in the open-label, parallel group study. Landiolol hydrochloride was given intravenously with a 1-minute loading infusion of 0.06 mg/kg/min, followed by a 60-minute infusion of 0.02 mg/kg/min using an automated infusion pump. Venous blood was drawn just before (predose) and 1, 2, 5, 15, 30 and 61 minutes after beginning the continuous intravenous infusion (during infusion); 2, 5, 10 and 30 minutes and 1, 4 and 8 hours after the end of the infusion (after infusion); and 24 hours after beginning the infusion (next day). Urine samples were collected up to 24 hours after beginning the infusion. Before subjects were discharged, an indocyanine green elimination test, clinical laboratory testing, physical examination and recording of ECGs and vital signs were performed. RESULTS: The geometric mean maximum plasma concentration and area under the concentration-time curve values for the patients with hepatic impairment were 42% and 44% higher, respectively, than those observed for the healthy volunteers, indicating that hepatic impairment affected the disposition of landiolol hydrochloride. There were no significant changes in the elimination half-life of the drug. There were no clinically significant differences between the two groups in terms of reductions in heart rate or blood pressure. CONCLUSION: The pharmacokinetic and pharmacodynamic characteristics of this ultra-short-acting beta1-blocker were maintained even in the patients with hepatic impairment. Although we did not observe any drug-related adverse events in these patients, hypotension or bradycardia should be considered, necessitating continuous monitoring of both heart rate and BP in patients with hepatic impairment who receive landiolol hydrochloride. PMID- 16274262 TI - Influence of silymarin administration on hepatic glutathione-conjugating enzyme system in rats treated with antitubercular drugs. AB - OBJECTIVE: This study evaluated the influence of simultaneous administration of silymarin (SIL), a hepatoprotective and antioxidant agent, on the status of glutathione (GSH) and its metabolising enzymes in the liver tissue of rats treated with antitubercular drugs, i.e. isoniazid (INH), rifampicin (RIF) and pyrazinamide (PYR). METHODS: Male Wistar albino rats (n = 24) were randomly divided into four groups. Group I received saline as they served as controls. Group II rats were administered antitubercular drugs (INH 25 mg/kg + RIF 50 mg/kg + PYR 140 mg/kg orally) daily for 45 days. Group III animals were treated with SIL (50 mg/kg orally) simultaneously with the antitubercular drugs for the same period. Group IV animals were treated with SIL alone. The status of GSH, glutathione peroxidase (GPx), glutathione reductase (GR) and glutathione-s transferase (GST) in liver tissue was evaluated at the end of the study. RESULTS: Administration of antitubercular drugs caused a significant decrease (p < 0.001) in the status of GPx, GST and GR and of non-enzymic (GSH) antioxidants in liver tissue when compared with saline-treated control rats. Simultaneous treatment of SIL with antitubercular drugs completely prevented decreases in the levels of all the above parameters. Treatment with SIL alone enhanced the activities of GST (p < 0.001) and GPx (p < 0.05) and did not alter glutathione levels compared with control. CONCLUSION: A fall in the status of glutathione and its conjugating enzymes upon administration of antitubercular drugs denotes an impairment of the antioxidant defence mechanism. Simultaneous administration of SIL afforded complete protection of the liver against this abnormality, an effect that could have been due to the strong antioxidant properties of SIL. PMID- 16274265 TI - Advanced magnetic resonance imaging techniques to evaluate CNS glioma. AB - To make full use of the benefits of magnetic resonance imaging (MRI) in the evaluation of intracranial glioma, a variety of advanced MRI and qualitative and quantitative techniques can be added to the radiologists' diagnostic armamentarium, beyond the standard contrast-enhanced images. These techniques include perfusion MRI, which is useful in the characterization of glioma because a relative quantification of cerebral microcirculatory parameters may be estimated; magnetic resonance spectroscopy, which can provide an estimate of the concentration of normal and abnormal metabolites in the brain and, when supplemented with magnetic resonance perfusion data, can be a useful tool for distinguishing between tumor recurrence and radiation necrosis; and diffusion tensor imaging, which combines physiologic behaviors unique to the cytoarchitecture of brain cells and the interstitial tissues with 2D and 3D anatomic magnetic resonance images, and thereby differentiates intact white matter tracts, displaced by tumor or surrounding edema, from invasion or disruption by a more aggressive neoplasm. PMID- 16274264 TI - Brain tumors: controversies and challenges in management. PMID- 16274263 TI - Ancient and recent positive selection transformed opioid cis-regulation in humans. AB - Changes in the cis-regulation of neural genes likely contributed to the evolution of our species' unique attributes, but evidence of a role for natural selection has been lacking. We found that positive natural selection altered the cis regulation of human prodynorphin, the precursor molecule for a suite of endogenous opioids and neuropeptides with critical roles in regulating perception, behavior, and memory. Independent lines of phylogenetic and population genetic evidence support a history of selective sweeps driving the evolution of the human prodynorphin promoter. In experimental assays of chimpanzee-human hybrid promoters, the selected sequence increases transcriptional inducibility. The evidence for a change in the response of the brain's natural opioids to inductive stimuli points to potential human-specific characteristics favored during evolution. In addition, the pattern of linked nucleotide and microsatellite variation among and within modern human populations suggests that recent selection, subsequent to the fixation of the human-specific mutations and the peopling of the globe, has favored different prodynorphin cis regulatory alleles in different parts of the world. PMID- 16274266 TI - Surgery for low-grade gliomas: current evidence and controversies. AB - Despite decades of studies, the surgical management of cerebral low-grade gliomas remains quite controversial. Recommendations range from observation with biopsy to radical resection. This controversy arises from several factors, chief of which is the lack of prospective, randomized studies specifically addressing the issue of resection. Nevertheless, the available data, with its inherent inadequacies, must be used to help guide the care of these patients. This article reviews the current literature on the role of surgery for these tumors and attempts to put forward reasonable treatment options in an area where even the American Association of Neurological Surgeons can find no standard management to recommend aside from biopsy prior to the onset of therapy. PMID- 16274267 TI - Chemotherapy for low-grade gliomas. AB - Low-grade gliomas pose a difficult problem for the neuro-oncologist. More needs to be known about their natural history. In addition, the role and timing of radiotherapy and chemotherapy in the management of low-grade tumors is highly controversial. Most agree that physicians should recommend treatment in symptomatic cases or in instances of radiologic tumor progression. This review discusses some of the recent advances with respect to chemotherapy for low-grade gliomas, with a special emphasis on low-grade astrocytomas and oligodendrogliomas. PMID- 16274268 TI - Oligodendrogliomas: clinical significance of 1p and 19q chromosomal deletions. AB - Oligodendrogliomas are a distinct subgroup of brain tumors with a fairly favorable clinical prognosis. However, these tumor exhibit varying degrees of heterogeneity and their clinical behavior is therefore not always the same. For this reason, genetic markers have been developed to further guide the clinical treatment. One such marker, the 1p and 19q chromosomal deletions, has been well documented in a subset of patients with oligodendrogliomas. Most importantly, patients who exhibit these chromosomal deletions respond favorably to chemotherapy. This article reviews the evidence describing the association of such deletions with a favorable response to chemotherapy and improved prognosis of patients with oligodendrogliomas. In addition, attempts to elucidate the molecular mechanisms behind the biologic behavior of these tumors are further explored. PMID- 16274269 TI - Surgical management of high-grade gliomas. AB - High-grade gliomas, in particular anaplastic astrocytoma and glioblastoma multiforme, represent two of the most devastating forms of brain cancer. In spite of the poor prognosis, new treatments and emerging therapies are making an impact on this disease. This review discusses the role of the surgical management of high-grade gliomas and provides an overview of the currently available therapies which depend on surgical intervention. At the same time, cutting-edge clinical trials for patients with malignant brain tumors are reviewed to provide further insights into potential future therapies. PMID- 16274270 TI - Chemotherapy for malignant glioma. AB - Malignant gliomas comprise a small percentage of all cancers, but continue to cause disproportionate levels of morbidity and mortality. Despite decades of intensive effort from many disciplines--surgery, radiation oncology and medicine- they remain refractory to cure and, in most cases, even to prolonged treatment response. Comprehensive multidisciplinary treatment is well recognized as the optimal approach. While continued advances and refinement in both surgical and radiotherapy-based techniques are certain, medical therapies are expanding at a much more rapid rate. This is due, in large part, to an understanding of the molecular events that underlie cancer pathogenesis and improved laboratory techniques to manufacture and study molecules that influence this process. This review will focus on medical therapies in the treatment of malignant glioma, never losing sight of their place as one of several therapeutic modalities used to confront brain cancer. PMID- 16274271 TI - Role of radiotherapy in the treatment of gliomas. AB - Malignant gliomas are challenging tumors that are often treated with a multimodality approach. This article focuses on the role of radiotherapy in the management of these tumors. The role of radiotherapy in low-grade gliomas remains controversial and this review focuses on the importance of prognostic factors, recent randomized trials involving radiotherapy, and toxicity from radiotherapy. In terms of high-grade gliomas, radiotherapy has a more established role and this review will address methods that have been evaluated in order to improve radiotherapy outcome. Improvements in radiotherapy delivery, tumor imaging and biologic modifiers may ultimately lead to improved outcome in the treatment of these difficult tumors. PMID- 16274272 TI - Primary central nervous system lymphoma. AB - Primary central nervous system lymphoma is a stage 1E non-Hodgkin's lymphoma confined to the nervous system. It is seen in immunocompetent and immunodeficient populations, the latter group associated with the Epstein-Barr virus. Primary central nervous system lymphoma can affect the brain, leptomeninges, spinal cord or eyes. The institution of high-dose methotrexate-based regimens and whole-brain radiation therapy has significantly increased survival, but neurotoxicity is high in patients over 60 years of age. Despite these advances, 50% of patients initially treated will relapse. Recent investigations include the use of rituximab (immunotherapy) and stem-cell transplantation, as well as regimens without whole-brain radiation therapy in the elderly. The optimal treatment regimen is yet to been determined. PMID- 16274273 TI - Managing symptoms and side effects during brain tumor illness. AB - Malignant brain tumors and the therapies used to treat them can present challenging problems. Headache is the most common symptom during brain tumor illness. Etiology determines the exact management approach, but pharmacologic and non pharmacologic measures may be used. Seizures also commonly occur and are best managed with anti epileptic drug therapy. Infection and deep venous thrombosis are concerns and are best approached by preventive measures and early aggressive intervention if those measures fail. Depression, fatigue, memory and personality changes may complicate care and are approached on an individual basis. Early discussion about end-of-life issues is necessary because the disease itself can impair decision-making ability. PMID- 16274277 TI - The children in the community study of developmental course of personality disorder. AB - The Children in the Community (CIC) Study is an ongoing investigation of the course of psychiatric disorders including personality disorders (PDs) in an epidemiological sample of about 800 youths. In addition to tracking developmental trajectories over 20 years from adolescence into adulthood, the CIC Study has used prospective data to investigate early risks for Axis II disorders and symptoms (including both environmental factors and early characteristics), implications of comorbidity with Axis I disorders, and associated negative prognostic risk of adolescent PDs into adulthood. The substantial independent impact of PD on subsequent Axis I disorders, suicide attempts, violent and criminal behavior, interpersonal conflict, and other problematic adult outcomes confirms the importance of attention to these problems when they manifest in early adolescence. The implications of study findings for potential changes in the DSM are discussed. PMID- 16274279 TI - The McLean Study of Adult Development (MSAD): overview and implications of the first six years of prospective follow-up. AB - The McLean Study of Adult Development (MSAD) began 12 years ago. It is the first NIMH-funded prospective study of the course and outcome of borderline personality disorder (BPD). After careful analysis of the first six years of follow-up, 5 main findings concerning the symptomatic and psychosocial course of BPD have emerged from this study. The first finding is that remissions are far more common than previously recognized (about 74%). The second is that these remissions are quite stable and thus, recurrences are quite rare (about 6%). The third finding is that completed suicides are far more rare than anticipated (about 4% vs. 10%). The fourth finding is that a "complex" model of borderline psychopathology best describes BPD. In this model, some symptoms resolve relatively quickly, are the best markers for the disorder, and are often the immediate reason for needing costly forms of treatment, such as psychiatric hospitalizations. We termed these symptoms (e.g., self-mutilation, help-seeking suicide threats or attempts) acute symptoms. Other symptoms resolve more slowly, are not specific to BPD, and are closely associated with ongoing psychosocial impairment. We termed such symptoms (e.g., chronic feelings of intense anger, profound abandonment concerns) temperamental symptoms. Fifth, it was also found that borderline patients were improving psychosocially over time, particularly remitted borderline patients; psychosocial functioning of remitted patients continued to improve as time progressed, suggesting that they were somewhat belatedly achieving the milestones of young adulthood and not simply returning to a prodromal level of functioning. Taken together, these results suggest that the prognosis for BPD is better than previously recognized. PMID- 16274280 TI - Stability and change in personality pathology: revelations of three longitudinal studies. AB - Three major longitudinal studies of personality disorder jointly provide new insights into the nature of stability and change in this important domain of psychopathology. This commentary highlights some of the many convergent results, which indicate robust findings that can be considered "established," and also differences in the studies' purposes and methods that permit each study to make unique contributions. One of the most important findings across the studies is that DSM- diagnosed personality disorder is not as stable as the characteristic traits that underlie personality pathology, leading to the insight that DSM personality disorders are hybrids of more acute, "Axis I-like" symptoms that resolve more quickly and longer lasting affective, cognitive, and behavioral personality dysfunctions. Implications for future research are considered. PMID- 16274278 TI - The Collaborative Longitudinal Personality Disorders Study (CLPS): overview and implications. AB - The Collaborative Longitudinal Personality Disorders Study (CLPS; Gunderson et al., 2000) was developed to fill gaps in our understanding of the nature, course, and impact of personality disorders (PDs). Here, we review published findings to date, discuss their implications for current conceptualizations of PDs, and raise questions that warrant future consideration. We have found that PDs are more stable than major depressive disorder, but that meaningful improvements are possible and not uncommon. We have confirmed also that PDs constitute a significant public health problem, with respect to associated functional impairment, extensive treatment utilization, negative prognostic impact on major depressive disorder, and suicide risk. At the same time, we have demonstrated that dimensional models of PDs have clinical validity that categories do not, especially greater temporal stability. Furthermore, dimensional personality traits appear to be the foundation of behaviors described by many PD criteria. Taken together, our results lead us to hypothesize that PDs may be reconceptualized as hybrids of stable personality traits and intermittently expressed symptomatic behaviors. PMID- 16274281 TI - Lessons from longitudinal studies for new approaches to the DSM-V: the FFM and FFT. AB - After brief comments about each target article, we discuss their significance for the DSM-V, the implications for personality disorders of universal trait developmental trends, and our emerging theoretical model, the Five-Factor Theory, which provides an integrative context for these remarkable findings. PMID- 16274282 TI - Progress and innovation: personality disorders and the vanguard of psychopathology research. AB - Important progress in our understanding of the natural course of personality disorders (PDs) is documented in the articles for this special section. This progress could set the stage for ideas developed in the study of PDs to play a central role in research on psychopathology more broadly conceived. The Collaborative Longitudinal Personality Disorders Study (Skodol et al., this issue), the Children in the Community Study (Cohen, Crawford, Johnson, & Kasen, this issue), and the McLean Study of Adult Development (Zanarini, Frankenburg, Hennen, Reich, & Silk, this issue) reveal the importance of personality in understanding psychopathology, and point toward a dimensional approach to conceptualizing psychopathology that could also frame categorical clinical decision making processes. PMID- 16274283 TI - Longitudinal studies of personality disorders: four lessons from personality psychology. AB - The three longitudinal projects described in this special section of the Journal of Personality Disorders raise a number of intriguing questions concerning the natural history of personality disorders and offer more than their share of surprises. In addition, they underscore several valuable lessons derived from the literature on normal-range personality traits. Drawing in part from the writings of the American trait psychologist Gordon Allport, I describe four such lessons: (1) change and continuity of personality traits and disorders can and do coexist, (2) the covariation among personality traits helps to account for the "comorbidity" among personality disorders, (3) personality traits and disorders influence how individuals interpret life events, and (4) personality traits must be distinguished from behavioral adaptations to these traits. These lessons remind us that the science of personality disorders must be informed by the basic science of personality. PMID- 16274284 TI - Outcome and epidemiological research on personality disorders: implications for classification. AB - The construct of personality disorder has an overall definition that describes chronicity. However, the findings of outcome research indicate that while many patients stop meeting criteria for specific categories of disorder over time, they often continue to suffer from dysfunctional traits. One can also interpret epidemiological findings on personality disorders as overestimating disorders due to a failure to distinguish them from traits. Taken together, these findings point to the need for a revision of the classification of personality disorders that would separate enduring traits from reversible symptoms. PMID- 16274285 TI - Changing views about personality disorders: Comment about the prospective studies CIC, CLPS, and MSAD. AB - Three large-scale prospective studies (Children in the Community Study [CIC; Cohen, Crawford, Johnson, & Kasen, this issue], Collaborative Longitudinal Personality Disorders Study [CLPS; Skodol et al., this issue], and the McLean Study of Adult Development [MSAD; Zanarini, Frankenburg, Hennen, Reich, & Silk, this issue]) are discussed with respect to the following issues: shared and complementary features of the study designs, evidence for stable and changing personality disorder features, methodological issues (reliability, stability, and invariance), the relationship between Axis I and II disorders, and dimensional versus categorical representations of personality disorders. PMID- 16274286 TI - Temporal change: the third dimension of personality disorder. AB - Until fairly recently, almost everyone who learned about, or carried out research into, personality disorders was taken in by the notion that personality disorder was a stable entity whose main characteristic was persistence, at least between the time of adolescence to middle age. We also suspected that those with these disorders, unless they received specific and necessarily powerful interventions, would remain continuously handicapped. Now it is clear that change in personality is possible, often rapidly so in some groups, and undergoes dynamic development from its genesis in childhood onward, and, though often achieving equilibrium, can never be said to be fixed in its manifestations. Longitudinal studies also show that personality abnormality identified early in life, whether temporary or longer-lasting, has a major influence on mental development and disorder generally, and, while it is impossible to say at present whether this influence is causal, it does suggest that early intervention could confer long-term benefits. An initiative such as that currently developed for early intervention in psychosis is at least as apt for personality disorder. PMID- 16274287 TI - Taxonomic implications of three prospective studies on the course and outcome of personality disorders. AB - The Children in the Community (CIC) Study (Cohen, Crawford, Johnson, & Kasen, this issue), the McLean Study of Adult Development (MSAD; Zanarini, Frankenburg, Hennen, Reich, & Silk, this issue), and the Collaborative Longitudinal Personality Disorders Study (CLPS; Skodol et al., this issue) are three large prospective studies on the course and outcome of psychiatric disorders, borderline personality disorder, and personality disorders, respectively. These studies, involving approximately 2,000 subjects, have yielded an enormous amount of scientifically and clinically relevant information. In this commentary, I primarily aim to discuss the implications for future revisions to the DSM classification of personality disorders. PMID- 16274288 TI - CIC, CLPS, and MSAD. AB - The purpose of this article is to provide a commentary on the reports of the findings from the longitudinal Children in the Community (CIC) study by Cohen, Crawford, Johnson, and Kasen (this issue), Collaborative Longitudinal Personality Disorders Study (CLPS) by Skodol et al. (this issue), and McLean Study of Adult Development (MSAD) by Zanarini, Frankenburg, Hennen, Reich, and Silk (this issue). This article focuses in particular on the implications of their longitudinal findings with regard to a fundamental component of personality disorder: temporal stability. Also discussed are their findings with regard to an integration of general personality structure with personality disorder. PMID- 16274290 TI - Functional dissection of the HNF-1alpha transcription factor: a study on nuclear localization and transcriptional activation. AB - Hepatocyte nuclear factor-1alpha (HNF-1alpha) is a homeodomain-containing transcription factor regulating the expression of liver and pancreas-specific genes. Mutations in the HNF-1alpha-encoding gene TCF1 cause maturity-onset diabetes of the young, type 3 (MODY3). These mutations may affect nuclear import or reduce the ability of HNF-1alpha to stimulate transcription. We performed a functional dissection of HNF-1alpha, attempting both to define its nuclear localization signals (NLSs) and to identify important elements of the Cterminal transactivation domain. Three HNF-1alpha regions, A (amino acids 158-171), B (197 205), and C (271-282), highly similar to consensus NLSs, were studied by immunolocalization in HeLa cells. Region B could be identified as the most critical for correct nuclear localization. Deletion of two subregions (amino acids 398-470 and 544-631, respectively) in the HNF-1alpha C-terminal transactivation domain, resulted in the greatest reduction in stimulation of transcription compared to wild-type protein. However, this domain probably consists of many elements that work in concert to give the full transactivation potential of the protein. PMID- 16274291 TI - In vitro plant regeneration and genetic transformation of Dichanthium annulatum. AB - Optimization of in vitro plant regeneration and genetic transformation of apomictic species such as Dichanthium annulatum would enable transfer of desirable genes. Seven genotypes of this grass species were screened through mature seed explant for embryogenic callus induction, callus growth and quality (color and texture), and shoot induction. Genotype IG-1999, which produced highly embryogenic, rapidly growing good-quality callus capable of regenerating at a high frequency, was selected for transformation experiments. Using a binary vector (pCAMBIA1305), frequency of GUS expression was compared between two methods of transformation. Bombardment of embryogenic calli with gold particles coated with pCAMBIA1305 at a distance of 11 cm, pressure of 4 bars, and vacuum of 27 Hg passing through 100 muM mesh produced maximum GUS expression (23%). Agrobacterium infection was maximum at an optical density of 2.0 when cocultured under vacuum for 15 min and cocultivated for 3 days at 28 degrees C in constant dark on MS medium of pH 5.8 with 3 mg/l 2,4-D, and 400 muM acetosyringone. Among two binary vectors used for Agrobacterium-mediated transformation, pCAMBIA1301 showed higher frequency of GUS expression while pCAMBIA1305 recorded more of the GUS spots per callus. Supplementation of acetosyringone in the cocultivation medium was found indispensable for Agrobacterium-mediated transformation. Injuring the calli through gold particle bombardment before their cocultivation with Agrobacterium improved the transformation efficiency. Several transgenic plants were developed using the PIG method, while stable GUS-expressing calli were multiplied during selection on MS medium containing 250 mg/l cefotaxime and 50 mg/l hygromycin, incubated in constant dark. A highly significant difference was observed between two methods of transformation for both frequency of GUS expression and GUS spots per callus. PIG-mediated transformation resulted in higher GUS expression compared to the Agrobacterium method. These results demonstrate that Dichanthium annulatum is amenable to Agrobacterium-mediated genetic transformation using a binary vector. PMID- 16274292 TI - Spontaneously immortalized mouse embryo fibroblasts: growth behavior of wild-type and vimentin-deficient cells in relation to mitochondrial structure and activity. AB - Dependent on the presence or absence of vimentin, primary mouse embryo fibroblasts exhibit different growth characteristics in vitro. While most Vim(+/+) fibroblasts stop dividing and die via apoptosis, a substantial fraction of cells immortalize and proliferate almost normally. Vim(-/-) fibroblasts cease to divide earlier, immortalize in vanishingly small numbers and thereafter proliferate extremely slowly. Early after immortalization, Vim(+/+) (imm) fibroblasts appear structurally almost normal, whereas Vim(-/-) (imm) fibroblasts equal postmitotic "crisis" cells, which are characterized by increased cell size, altered cell ultrastructure, nuclear enlargement, genome destabilization, structural degeneration of mitochondria, and diminution of mitochondrial respiratory activity. The differences between immortalized Vim(+/+) (imm) and Vim(-/-) (imm) fibroblasts persist during early cell cloning but disappear during serial subcultivation. At high cell passage, cloned, immortalized vim(-) fibroblasts grow nearly as fast as their cloned vim(+) counterparts, and also resemble them in size, ultrastructure, nuclear volume, and mitochondrial complement; they very likely employ redundancy to cope with the loss of vimentin function when adjusting structure and behavior to that of immortalized vim(+) fibroblasts. Reduction in nuclear size occurs via release of large amounts of filamentous chromatin into extracellular space; because it is complexed with extracellular matrix proteins, it tends to form clusters and to tightly stick to the surface of other cells, thus providing a potential for horizontal gene transfer. On the other hand, cloned vim(+) and vim(-) fibroblasts are equal in showing contact inhibition at young age and becoming anchorage-independent during serial subcultivation, as indicated by the formation of multilayered and -faceted cell sheets and huge spheroids on top of or in soft agar. With this, immortalized vim(-) fibroblasts reduce their adhesiveness to the substratum which, in their precrisis state and early after cloning, is much higher than that of their vim(+) counterparts. In addition, the coupling between the mitochondrial respiratory chain and oxidative phosphorylation is stronger in vim(+) than vim(-) fibroblasts. It appears from these data that after explantation of fibroblasts from the mouse embryo the primary cause of cell and mitochondrial degeneration, including genomic instability, is the mitochondrial production of reactive oxygen species in a vicious circle, and that vimentin provides partial protection from oxidative damage. As a matrix protein with specific in vitro and in vivo affinities for nuclear and mitochondrial, recombinogenic DNA, it may exert this effect preferentially at the genome level via its influence on recombination and repair processes, and in this way also assist the cells in immortalizing. Additional protection of mitochondria by vimentin may occur at the level of mitochondrial fatty acid metabolism. PMID- 16274293 TI - Stability and association with the cytomatrix of mitochondrial DNA in spontaneously immortalized mouse embryo fibroblasts containing or lacking the intermediate filament protein vimentin. AB - To extend previous observations demonstrating differences in number, morphology, and activity of mitochondria in spontaneously immortalized vim(+) and vim(-) fibroblasts derived from wild-type and vimentin knockout mice, some structural and functional aspects of mitochondrial genome performance and integrity in both types of cells were investigated. Primary Vim(+/+) and Vim(-/-) fibroblasts, which escaped terminal differentiation by immortalization were characterized by an almost twofold lower mtDNA content in comparison to that of their primary precursor cells, whereby the average mtDNA copy number in two clones of vim(+) cells was lower by a factor of 0.6 than that in four clones of vim(-) cells. However, during serial subcultivation up to high passage numbers, the vim(+) and vim() fibroblasts increased their mtDNA copy number 1.5- and 2.5-fold, respectively. While early-passage cells of the vim(+) and vim(-) fibroblast clones differed only slightly in the ratio between mtDNA content and mitochondrial mass represented by mtHSP70 protein, after ca. 300 population doublings the average mtDNA/mtmass ratio in the vim(+) and vim() cells was increased by a factor of 2 and 4.5, respectively. During subcultivation, both types of cells acquired the fully transformed phenotype. These findings suggest that cytoskeletal vimentin filaments exert a strong influence on the mechanisms controlling mtDNA copy number during serial subcultivation of immortalized mouse embryo fibroblasts, and that vimentin deficiency causes a disproportionately enhanced mtDNA content in high-passage vim(-) fibroblasts. Such a role of vimentin filaments was supported by the stronger retention potential for mtDNA and mtDNA polymerase (gamma) detected in vim(+) fibroblasts by Triton X-100 extraction of mitochondria and agaroseembedded cells. Moreover, although the vim(+) and vim(-) fibroblasts were equally active in generating free radicals, the vim(-) cells exhibited higher levels of immunologically detectable 8-oxoG and mismatch repair proteins MSH2 and MLH1 in their mitochondria. Because in vim(-) fibroblasts only one point mutation was detected in the mtDNA D-loop control region, these cells are apparently able to efficiently remove oxidatively damaged nucleobases. On the other hand, a number of large-scale mtDNA deletions were found in high-passage vim(-) fibroblasts, but not in low-passage vim(-) cells and vim(+) cells of both low and high passage. Large mtDNA deletions were also induced in young vim(-) fibroblasts by treatment with the DNA intercalator ethidium bromide, whereas no such deletions were found after treatment of vim(+) cells. These results indicate that in immortalized vim(-) fibroblasts the mitochondrial genome is prone to large-scale rearrangements, probably due to insufficient control of mtDNA repair and recombination processes in the absence of vimentin. PMID- 16274294 TI - Diversity and complexity of the mu opioid receptor gene: alternative pre-mRNA splicing and promoters. AB - Mu opioid receptors play an important role in mediating the actions of a class of opioids including morphine and heroin. Binding and pharmacological studies have proposed several mu opioid receptor subtypes: mu(1), mu(2), and morphine-6beta glucuronide (M6G). The cloning of a mu opioid receptor, MOR-1, has provided an invaluable tool to explore pharmacological and physiological functions of mu opioid receptors at the molecular level. However, only one mu opioid receptor (Oprm) gene has been isolated. Alternative pre-mRNA splicing has been proposed as a molecular explanation for the existence of pharmacologically identified subtypes. In recent years, we have extensively investigated alternative splicing of the Oprm gene, particularly of the mouse Oprm gene. So far we have identified 25 splice variants from the mouse Oprm gene, which are controlled by two diverse promoters, eight splice variants from the rat Oprm gene, and 11 splice variants from the human Oprm gene. Diversity and complexity of the Oprm gene was further demonstrated by functional differences in agonist-induced G protein activation, adenylyl cyclase activity, and receptor internalization among carboxyl terminal variants. This review summarizes these recent results and provides a new perspective on understanding and exploring complex opioid actions in animals and humans. PMID- 16274295 TI - Efficacy of combination gene therapy with multiple growth factor cDNAs to enhance skin flap survival in a rat model. AB - The objective of this study was to investigate the efficacy of combination gene therapy with multiple angiogenic growth factor cDNAs to enhance survival of ischemic skin flaps in a rat model. Sixty Sprague-Dawley rats were divided into six groups. Varying combinations of VEGF165, PDGF-B, and bFGF-plasmids were injected to prefabricate the flaps. Random skin flaps were raised on the dorsal aspect of rats following prefabrication with growth factor cDNAs. Flap viability was determined by measurement of percentage area of survival. The efficacy of gene therapy was evaluated by flap survival and neovascularization of representative histologic sections stained immunohistologically. The VEGF165 plus bFGF cDNAs enhanced the viability of the flap and neovascularization most effectively; the flap survival area was 64.3 +/- 8.7% after transfer of these two growth factor genes. Addition of PDGF-B cDNA is deleterious to the effects of combined VEGF165 and bFGF, leading to a significant decrease in flap viability (44.9 +/- 2.7%). Viability of the flaps with combined VEGF165 and bFGF cDNA transfer was significantly greater than that of the flaps with VEGF165 transfer alone (57.6 +/- 5.2%) or sham plasmid control (52.3 +/- 5.0%). Combined transfer of VEGF165 and bFGF cDNA is the most effective combination of multiple growth factor genes to improve flap viability in this model. Simultaneous transfer of three growth factor genes (VEGF165, PDGF-B, and bFGF) is deleterious to flap survival, at least for the ratio of lipofectin:transgene employed. PMID- 16274296 TI - Maternal immunization with both hemagglutinin- and neuraminidase-expressing DNAs provides an enhanced protection against a lethal influenza virus challenge in infant and adult mice. AB - Maternal immunization is the major form of protection against many infectious diseases in early life. In this report, transmission of vaccine-specific maternal antibodies and protection of offspring against a lethal influenza virus challenge were studied. Adult female BALB/c mice were immunized intramuscularly with plasmid DNAs encoding influenza virus hemagglutinin (HA), neuraminidase (NA), or mixture of the two plasmids. The levels of specific antibodies in sera of offspring at different ages and the survival rates following the lethal viral challenge were valued. The results showed effective transmission of maternal antibodies and long-lasting protection in offspring. Along with the growth of offspring, the antibody titers in vivo decreased and the ability against virus infection decreased accordingly. The HA-specific maternal antibodies protected the offspring from a lethal influenza infection up to 2 weeks old, and the NA specific maternal antibodies protected offspring up to 4 weeks old. Furthermore, antibodies transferred by the mother immunized with the mixture of HA and NA DNAs protected the offspring up to 6 weeks old. This suggests that maternal immunization with a mixture of HA and NA DNAs provide the most effective protection against the virus challenge for the offspring of mice. PMID- 16274297 TI - Expression of endothelins and their receptors promotes an invasive phenotype of breast tumor cells but is insufficient to induce invasion in benign cells. AB - There is increased staining of endothelins (ET-1, -2, and -3) and receptors (ET RA and -RB) in invasive breast tumors compared to nonneoplastic tissue, and ETs stimulate MCF-7 cell invasion in vitro. We analyzed ETstimulation of benign and transformed mammary epithelial cells, and whether expression of ETs is sufficient to induce invasiveness. In breast cancer patient serum, ET-1 was increased in those patients with lymph node metastases compared to those with no lymph node involvement; ETs, however, had no mitogenic effect on breast tumor cell lines in vitro. The benign mammary epithelial cell line, hTERT-HME1, and the poorly invasive breast tumor cell line MCF-7 secreted low levels of ET-1, while the invasive cell lines SKBR3 and MDAMB231 secreted high levels. Expression of the ETs and receptors by the cell lines broadly correlated with their in vitro invasiveness; overexpression of ETs in MCF-7 cells increased basal invasion. ET mediated invasion involved both receptors and a calcium influx to induce a pertussis toxin-sensitive MAPK pathway. MMP-14 activity was induced via ET-RA in an autocrine manner. In contrast to transformed cells, ET stimulation or overexpression did not induce an invasive phenotype in benign cells. Benign cells do not respond to ETs, and ET expression is not sufficient to induce invasion; however, the level of ET production by tumor cells correlates with their invasiveness, and increasing expression of the ET axis promotes breast tumor cell invasion via both receptors, while MMP-14 is induced via ET-RA. PMID- 16274298 TI - A systematic review on the effects of subgingival chlorhexidine gel administration in the treatment of chronic periodontitis. AB - BACKGROUND: Since scaling and root planing are not always successful, predominantly when access to periodontal defects is compromised, various antimicrobial agents as adjuncts have been suggested to enhance their efficacy. Gel vehicles delivering chlorhexidine (CHX) have become available and tested for these indications. METHODS: An electronic (MEDLINE and Cochrane Oral Health Group Specialized Trials Register) and manual search were made to detect studies concerning the use of chlorhexidine gels as a single measure of treatment and as an adjunct to scaling and root planing. Only full-text randomized controlled trials published in English up to January 2005 were included. RESULTS: Eight studies were finally selected. Due to considerable heterogeneity in study design and outcome variables measured a qualitative data analysis was performed. There is evidence that subgingival chlorhexidine gel administration as a monotherapy temporarily reduces bleeding tendency on probing; a clinical effect coinciding with relevant microbiological changes described in all but one study. To what extent chemical effects contribute to these changes appears to be related to the frequency of gel administration. There seem to be little to no data indicating that the treatment outcome of scaling and root planing will benefit from the adjunctive subgingival administration of a CHX gel. CONCLUSION: The limited data currently available on the effects of subgingival chlorhexidine gel application do not justify its use in the treatment of chronic periodontitis. PMID- 16274299 TI - Acellular dermal matrix for mucogingival surgery: a meta-analysis. AB - BACKGROUND: Many clinical studies revealed the effectiveness of acellular dermal matrix (ADM) in the treatment of mucogingival defects. The purpose of this meta analysis was to compare the efficacy of ADM-based root coverage (RC) and ADM based increase in keratinized tissues to other commonly used mucogingival surgeries. METHODS: Meta-analysis was limited to randomized clinical trials (RCT). Articles from January 1, 1990 to October 2004 related to ADM were searched utilizing the MEDLINE database from the National Library of Medicine, the Cochrane Oral Health Group Specialized Trials Registry, and through hand searches of reviews and recent journals. Relevant studies were identified, ranked independently, and mean data from each were weighted accordingly. Selected outcomes were analyzed using a meta-analysis software program. The significant estimates of the treatment effects from different trials were assessed by means of Cochrane's test of heterogeneity. RESULTS: 1) Few RCT studies were found to compile the data. In summary, selection identified eight RCT that met the inclusion criteria. There were four studies comparing ADM versus a connective tissue graft for root coverage procedures, two studies comparing ADM versus coronally advanced flap (CAF) for root coverage procedures, and two studies comparing ADM to free gingival graft in augmentation of keratinized tissue. 2) There were no statistically significant differences between groups for any of the outcomes measured (recession coverage, keratinized tissue formation, probing depths, and clinical attachment levels). 3) The majority of the analyses demonstrated moderate to high levels of heterogeneity. 4) Considering the heterogeneity values found among the studies, certain trends could be found: a) three out of four studies favored the ADM-RC group for recession coverage; b) a connective tissue graft tended to increase keratinized tissue compared to ADM (0.52-mm difference; P = 0.11); c) there were trends of increased clinical attachment gains comparing ADM to CAF procedures (0.56-mm difference; P = 0.16). CONCLUSIONS: Differences in study design and lack of data precluded an adequate and complete pooling of data for a more comprehensive analysis. Therefore, considering the trends presented in this study, there is a need for further randomized clinical studies of ADM procedures in comparison to common mucogingival surgical procedures to confirm our findings. It is difficult to draw anything other than tentative conclusions from this meta-analysis of ADM for mucogingival surgery, primarily because of the weakness in the design and reporting of existing trials. PMID- 16274300 TI - Histologic and histomorphometric findings from retrieved, immediately occlusally loaded implants in humans. AB - BACKGROUND: The immediate loading treatment concept can be successfully used in implant dentistry. Bone cells migrate onto the implant surface and establish a stable anchorage on the titanium surface. When implants are loaded immediately after surgery, there is a high long-term success rate of the implant-supported reconstruction. Based on histologic observations from different animal studies, the interface of immediately loaded implants can have a direct bone-to-implant connection without any fibrous tissue formation. Mature bone formation is dependent on the loading period. The aim of this study was to demonstrate a histologic analysis of retrieved, clinically stable immediately loaded implants with different implant designs and surfaces. An objective demonstration of the bone-implant interface was presented for the implant systems used. METHODS: A total of 29 implants with different implant designs and surfaces were retrieved from patients who were treated with implants using an immediate loading protocol and fixed immediate restorations placed the same day after surgery. The loading period was between 2 and 10 months. The bone-implant interface was examined histologically and histomorphometrically. RESULTS: A high bone-to-implant percentage of 66.8% (+/-8.9%) was found in the examined retrieved implants. Some marginal bone resorption was observed in the crestal part of the implants. CONCLUSION: According to the present histologic and histomorphometric evaluation of retrieved, clinically stable implants, immediate occlusal loading can present a high level of bone-to-implant contact in humans. PMID- 16274301 TI - Platelet-derived growth factor enhancement of two alloplastic bone matrices. AB - BACKGROUND: The use of alloplastic matrices that mimic the mineral phase of bone has become a viable alternative to current mainstream therapies in dentistry such as allografts and autogenous grafts. Because alloplastic bone substitutes generally have relatively poor osteogenic properties, analyzing their potential as vehicles to deliver growth factors is an important step in assessing methods to enhance their clinical efficacy. The aim of these studies was to treat beta tricalcium phosphate (beta-TCP) and calcium sulfate (CaSO(4)) with platelet derived growth factor (PDGF)-BB to enhance the osteogenic capabilities of these materials. METHODS: In the beta-TCP studies, PDGF-BB adsorption and release were accomplished using (125)I radiolabeled growth factor and non-radioactive human recombinant PDGF at a ratio of 1:300 M. For the adsorption studies, the radiolabeled PDGF-BB/ non-radioactive PDGF solutions with resultant PDGF concentrations of 10(7) and 10(8) M were incubated with beta-TCP from 1 to 120 minutes, and the amount of adsorbed (125)I-PDGF-BB was measured using a gamma counter. Similar adsorption studies were conducted with a 30-minute incubation of beta-TCP with various PDGF concentrations. In vitro release studies were conducted with beta-TCP to which radiolabeled PDGF had been adsorbed as above. Release studies were also conducted with CaSO(4) that was hydrated with the radioactive PDGF solution described above for the TCP studies. In vivo PDGF-BB release from beta-TCP and CaSO(4) was evaluated in a mouse model, where the radioactive PDGF/non-radioactive PDGF-BB treated beta-TCP or CaSO(4) sample was inserted subcutaneously and later removed for radioactive measurement. Proliferation of human osteoblastic cells in the presence of PDGF- treated beta TCP or CaSO(4) was assessed by (3)H thymidine incorporation. RESULTS: The absorption studies revealed that PDGF-BB was absorbed in a concentration and time dependent manner to beta-TCP. In the in vitro release studies, approximately 45% of the adsorbed PDGF-BB was released after 10 days. In vivo release from both materials occurred faster than in vitro release. Osteoblastic cells incubated with PDGF-BB-treated matrices showed significantly (P <0.05, ANOVA) greater proliferation than with control matrices alone. CONCLUSION: These experiments demonstrate the feasibility of using PDGF-BB in combination with alloplastic materials such as beta-TCP or CaSO(4) to serve as more effective bone graft materials with enhanced osteogenic properties. PMID- 16274302 TI - Treatment of gingival recession: comparison of two techniques of subepithelial connective tissue graft. AB - BACKGROUND: The increasing interest in esthetics and the subsequent need to solve related problems such as hypersensitivity and root caries have favored the development of many surgical procedures that permit the coverage of exposed roots. This clinical study was conducted to examine the coverage of gingival recession defects, where two different subepithelial connective tissue graft (SCTG) techniques (Langer and Langer and modified tunnel) were used. METHODS: Thirty one patients (21 females and 10 males), each contributing Miller Class I and II gingival recessions, were selected. Recession defects were randomly treated by using the Langer and Langer technique (17 patients) or the modified tunnel technique (14 patients). Vertical recession, probing depth (PD), and attachment level were assessed at baseline and 6 months postoperatively. RESULTS: Six months after the surgery, a significant reduction in recession depth was noticed in both groups. There was also a decrease of PD and attachment level for both groups, but not statistically significant. In comparison, at 6 months, statically significant differences were found between the tunnel and Langer and Langer techniques for root coverage and attachment gain. The percentage of root coverage was 96.4% and 75.5% in the tunnel and Langer and Langer groups, respectively. CONCLUSIONS: The present study suggests that the use of SCTG in combination with a tunnel procedure may result in an increased amount of root coverage and clinical attachment gain compared to the Langer and Langer technique. Further comparative studies are necessary to understand the periodontal healing generated by the tunnel procedure and Langer and Langer technique. PMID- 16274303 TI - Gingival crevicular fluid levels of monocyte chemoattractant protein-1 and tumor necrosis factor-alpha in patients with chronic and aggressive periodontitis. AB - BACKGROUND: Monocyte chemoattractant protein-1 (MCP-1) is a well-known chemotactic cytokine that regulates mononuclear inflammatory cell recruitment. This recruitment has particular importance in the oral cavity because inflammatory cells will be challenged with periodontopathogenic bacteria during infections. Tumor necrosis factor-alpha (TNF-alpha) is a cytokine that induces bone resorption by stimulating proliferation and differentiation of osteoclasts' progenitors and also stimulates MCP-1 expression. The aims of this study were to investigate the presence of MCP-1 in gingival crevicular fluid (GCF) samples from patients with chronic periodontitis (CP) and aggressive periodontitis (AgP) and to examine the possible correlations between the GCF levels of MCP-1 and TNF alpha. METHODS: Twenty-five CP, 20 AgP, and 20 healthy control (C) patients were selected for the study. Plaque index (PI), gingival index (GI), probing depth (PD), and clinical attachment level (CAL) measurements were recorded from each sampling area. Assays for GCF MCP-1 and TNF-alpha were carried out by an enzyme linked immunosorbent assay (ELISA) method. RESULTS: The concentration (nanograms per microliters) and total MCP-1 and TNF-alpha (nanograms per site) were not statistically significant between CP and AgP groups, but total MCP-1 and TNF alpha was statistically different between CP and C and between AgP and C groups (P <0.001). All clinical parameters were statistically different between CP and C and between AgP and C groups (P <0.001). A positive statistical correlation was detected between the levels of MCP-1 and TNF-alpha, and there was also a positive correlation between all clinical parameters and total MCP-1 and TNF-alpha levels. CONCLUSIONS: These results suggest that MCP-1 could have an important role in the activation and recruitment of inflammatory and immune cells in periodontal diseases, and both AgP and CP patients may have the same pattern of MCP-1 expression. A strong positive correlation between the GCF levels of MCP-1 and TNF alpha may account for the mechanism of amplification of inflammatory events in gingival inflammation. PMID- 16274304 TI - Dimension of keratinized mucosa and the interproximal papilla between adjacent implants. AB - BACKGROUND: The interproximal papilla between two adjacent implants is gaining critical interest in the implant dentistry. The aims of this study were to evaluate the effect of 1) the width of keratinized mucosa, 2) the distance from the base of the contact point to the crestal bone, and 3) the horizontal distance between two implants on the radiographic dimension from the tip of the papilla to the crestal bone between two implants. METHODS: This study involved 72 interproximal papillae between two adjacent implants (interimplant papilla) in 52 patients who had implants placed adjacent to each other and had a prosthesis in place more than 1 year. The shortest distance between the radiopaque material on the tip of the interimplant papilla and most coronal portion of the interimplant crestal bone was measured (the radiographic length of the papilla [RL]). The width of keratinized mucosa from the tip of the papilla was measured (WK). The vertical distance between the base of the contact point and the interimplant crestal bone was measured (CC). The horizontal distance between the two adjacent implants was measured at the fixture-abutment interface level (HD). Multiple regression analysis was performed between WK-RL, CC-RL, and HD-RL. RESULTS: RL had a significant relation with WK between two adjacent implants (P = 0.001). However, RL was not related with other variables, such as CC and HD (CC, P = 0.641; HD, P = 0.901). CONCLUSION: The results of this study suggest that the width of keratinized mucosa between two adjacent implants might be related to the dimension of the interproximal papilla between two adjacent implants. PMID- 16274305 TI - Local simvastatin effects on mandibular bone growth and inflammation. AB - BACKGROUND: Simvastatin has been shown to increase bone growth when applied topically to murine bone; however, it causes considerable soft tissue inflammation at high doses (2.2 mg), making future clinical use problematic. This study evaluated the effect of lower simvastatin doses and cyclooxygenase (COX) synthase inhibitors on tissue inflammation and bone growth in rats and gene expression in mice. METHODS: Adult female rats were untreated or treated with a single dose of 0.1, 0.5, 1.0, 1.5, or 2.2 mg simvastatin in methylcellulose gel in a polylactic acid membrane (SIM) on the lateral aspect of the mandible. The contralateral mandible side was implanted with methylcellulose gel/polylactic acid membrane alone (GEL), and five rats in each dose pairing were evaluated histomorphometrically after 3, 7, and 24 days. Subsequent rats were similarly treated with 0.5 mg simvastatin (optimal dose) and daily intraperitoneal injections of COX-2 inhibitor (NS-398; 1 mg/kg x 7 days; N = 16), general COX inhibitor (indomethacin; 1 mg/kg x 7 days; N = 16), or no inhibitor (N = 10) and evaluated histomorphometrically after 7 or 24 days by analysis of variance (ANOVA). Gene arrays were also used to evaluate osteogenic gene expression from 0.5 mg simvastatin in murine calvaria (N = 12). RESULTS: There was a 45% increase in bone area with 0.5 mg simvastatin versus gel control (P <0.001; similar to the 2.2-mg dose), and clinical swelling was reduced compared to the high simvastatin dose (P <0.05). The 0.1-mg simvastatin dose failed to stimulate significant bone growth. NS-398 and indomethacin reduced inflammation and bone growth. Simvastatin significantly upregulated procollagen, fibronectin, and matrix metalloproteinase 13 genes. CONCLUSION: Reducing the simvastatin dose from 2.2 to 0.5 mg reduced inflammation to a more clinically acceptable level without sacrificing bone growth potential, but COX-associated inflammation appears to be necessary for in vivo bone growth. PMID- 16274307 TI - Plasma levels of mannan-binding lectin in relation to periodontitis and smoking. AB - BACKGROUND: Mannan-binding lectin (MBL) is an important molecule of innate immunity; it acts as an opsonin and stimulates the classical complement pathway. Moreover, it has been suggested that MBL acts as a weak acute phase protein. We investigated whether MBL levels are increased in periodontitis, and we tested whether individuals deficient for MBL are more susceptible to periodontitis. METHODS: A total of 219 subjects participated in the study. Plasma samples from 115 periodontitis patients and 104 healthy controls were taken, and the MBL levels were measured by enzyme-linked immunosorbent assay (ELISA). MBL levels were analyzed in relation to periodontitis, taking into consideration age, gender, ethnic and educational background, and smoking status. In some analyses, subjects with MBL plasma levels <0.8 microg/ml were considered MBL deficient. RESULTS: MBL plasma concentrations were not significantly different in moderate and severe periodontitis compared to controls (1.6, 1.4, and 1.6 microg/ml, respectively). Also, the prevalence of MBL deficiency was not found to be different between controls and moderate and severe periodontitis (45%, 37%, and 36%). However, among all subjects and among the non-deficient subjects, MBL levels were markedly increased in heavy smokers (>10 cigarettes per day), irrespective of periodontal disease status, in comparison to non-smokers and light smokers. MBL plasma levels did not show a correlation with plasma C reactive protein (CRP) and were also not related to the prevalence of specific periodontal pathogens. CONCLUSIONS: MBL levels were not elevated in periodontitis, and MBL deficiency was not related to susceptibility for periodontitis. The fact that MBL levels were higher among heavy smokers is the subject of further investigation. PMID- 16274306 TI - Coverage of Miller class I and II recession defects using enamel matrix proteins versus coronally advanced flap technique: a 2-year report. AB - BACKGROUND: The aim of this study was to evaluate a comparison of the coronally advanced flap procedure with or without the use of enamel matrix proteins in the treatment of recession defects. METHODS: This 2-year study was conducted as a blinded, split-mouth, placebo-controlled, and randomized design. Thirty patients from two dental schools with two paired buccal recession defects were chosen. Surgical recession coverage was performed as the coronally advanced flap technique. One site was additionally treated with derivative (EMD) and the other site with a placebo (propylene glycol alginate [PGA]). A blinded examiner assessed pre- and post-surgical measurements. Measurements comprised the height and width of the gingival recession, height of keratinized tissue, probing attachment level, probing depth, and alveolar bone level. RESULTS: Twenty-four months after therapy, both treatment modalities showed significant root coverage and probing attachment gain. The mean gingival recession decreased from 3.6 to 0.8 mm for the EMD-treated sites and from 3.8 to 1.4 mm for the control sites. However, this difference was not statistically significant (P = 0.122). Similarly, all other clinical parameters did not differ significantly in the between-group comparison except for the recession width (P = 0.027) and probing depth (P = 0.046) exhibiting higher reductions in the EMD group. Complete root coverage could be maintained over 2 years in 53% of the EMD versus merely 23% in the control group. A total of 47% of the treated recessions in the control group deteriorated again in the second year after therapy compared to 22% in the EMD group. CONCLUSION: Enamel matrix derivative seems to provide better long-term results. PMID- 16274308 TI - Bisphosphonate modulates cementoblast behavior in vitro. AB - BACKGROUND: Cementum formation is deemed to be instrumental for the successful regeneration of periodontal tissues, and thus events and modifiers of cementum formation and mineralization need to be determined. This study aimed to determine whether the bisphosphonate 1-hydroxyethylidene-1,1-bisphosphonate (HEBP) altered the behavior of immortalized cementoblasts (osteocalcin-cementoblasts [OCCM]). METHODS: OCCM from transgenic mice were exposed to HEBP at concentrations ranging from 0.01 to 10.0 microM. The assays performed included the count of cell number for proliferation, Northern blot analysis for gene expression (up to 10 days for core binding factor alpha-1 [Cbfa1], bone sialoprotein [BSP], osteocalcin [OCN], and osteopontin [OPN], markers for cementoblast/osteoblast maturation/mineralization), von Kossa stain and alizarin red S stain for mineralization, and enzyme assay (p-nitrophenol phosphate cleavage) for alkaline phosphatase (ALP) activity. RESULTS: Mineral nodule formation was inhibited at the higher doses of HEBP (1.0 and 10.0 microM) only. At early stages (1, 3, and 6 days), gene expression assays revealed only subtle changes in treated cells versus untreated cells, but by day 10, groups treated with lower doses (0.01 and 0.1 microM) were markedly different at the gene expression level. OCN was significantly downregulated (70%) at the lowest dose, with less pronounced effects at higher doses. In concurrence, the master switch gene for osteoblasts, Cbfa1, was also downregulated at the lower doses. Inversely, OPN mRNA was enhanced at the lower doses. ALP activity was not altered by HEBP. CONCLUSION: Bisphosphonate alters cementoblast function in vitro through the regulation of gene expression and mineral formation. PMID- 16274309 TI - Effect of sodium alendronate on alveolar bone resorption in experimental periodontitis in rats. AB - BACKGROUND: Bisphosphonates are potent inhibitors of bone resorption and were shown to inhibit bone resorption in experimental periodontitis by unknown mechanisms. We studied the effect of the aminobisphosphonate sodium alendronate (SA) in experimental periodontitis. Wistar rats were subjected to ligature placement around the second upper left molars. METHODS: Animals were treated with SA 0.01 to 0.25 mg/kg subcutaneously (sc), either 1 hour before (prophylactic) or starting 5 days after (therapeutic) periodontitis induction and daily until the rats were sacrificed (11 days). Controls received saline. Animals were weighed daily. Alveolar bone loss was measured as the difference (in millimeters) between the cusp tip and the alveolar bone. The periodontium and the surrounding gingivae were examined at histopathology, and the neutrophil influx into the gingivae was assayed using myeloperoxidase activity. The local bacterial flora was assessed through culture of the gingival tissue in standard aerobic and anaerobic media. RESULTS: Alveolar bone loss was significantly and dose dependently inhibited by SA either as a prophylactic or therapeutic treatment compared to the control. SA reduced tissue lesion at histopathology, with partial preservation of the periodontium, coupled to decreased myeloperoxidase activity compared to the control. The reduced neutrophil influx was also shown in carrageenan-induced peritonitis, used as a control experiment for this parameter. SA also significantly inhibited the growth of pigmented bacilli and Fusobacterium nucleatum, which are important in the pathogenesis of periodontal disease. SA also inhibited the in vitro growth of isolated Peptostreptococcus sp. CONCLUSION: Sodium alendronate preserves alveolar bone resorption and has anti-inflammatory and antibacterial activities in experimental periodontitis. PMID- 16274310 TI - Risk indicators for tooth loss due to periodontal disease. AB - BACKGROUND: Several risk indicators for periodontal disease severity have been identified. The association of these factors with tooth loss for periodontal reasons was investigated in this cross-sectional comparative study. METHODS: All extractions performed in 21 general dental practice clinics (25% of such clinics in Kuwait) over a 30-day period were recorded. Documented information included patient age and gender, medical history findings, dental maintenance history, toothbrushing frequency, types and numbers of extracted teeth, and the reason for the extraction. Reasons were divided into periodontal disease versus other reasons in univariate and binary logistic regression analyses. RESULTS: A total of 1,775 patients had 3,694 teeth extracted. More teeth per patient were lost due to periodontal disease than for other reasons (2.8 +/- 0.2 versus 1.8 +/- 0.1; P <0.001). Factors significantly associated with tooth loss due to periodontal reasons in logistic regression analysis were age >35 years (odds ratio [OR] 3.45; 95% confidence interval [CI] 2.79 to 4.26), male gender (OR 1.42; 95% CI 1.17 to 1.73), never having periodontal maintenance (OR 1.48; 95% CI 1.23 to 1.78), never using a toothbrush (OR 1.81; 95% CI 1.49 to 2.20), current or past smoking (OR 1.56; 95% CI 1.28 to 1.91), anterior tooth type (OR 3.23; 95% CI 2.57 to 4.05), and the presence of either of the following medical conditions: diabetes mellitus (OR 2.64; 95% CI 2.19 to 3.18), hypertension (OR 1.73; 95% CI 1.41 to 2.13), or rheumatoid arthritis (OR 4.19; 95% CI 2.17 to 8.11). CONCLUSION: Tooth loss due to periodontal disease is associated with the risk indicators of age, male gender, smoking, lack of professional maintenance, inadequate oral hygiene, diabetes mellitus, hypertension, rheumatoid arthritis, and anterior tooth type. PMID- 16274311 TI - Tobacco smoking and periodontal health in a Saudi Arabian population. AB - BACKGROUND: The objective of this study was to examine the association between tobacco smoking, in particular water pipe smoking, and periodontal health. METHODS: A total of 262 citizens of Jeddah, Saudi Arabia in the age range from 17 to 60 years volunteered to participate in the study. The clinical examinations were carried out at King Faisal Specialty Hospital and Research Center in Jeddah and included assessments of oral hygiene, gingival inflammation, and probing depth. Smoking behavior was registered through a questionnaire and confirmed by an interview. Participants were stratified into water pipe smokers (31%), cigarette smokers (19%), mixed smokers (20%), and non-smokers (30%). RESULTS: The mean probing depth per person was 3.1 mm for water pipe smokers, 3.0 mm for cigarette smokers, 2.8 mm for mixed smokers, and 2.3 mm for non-smokers. The association between smoking and probing depth was statistically significant controlling for age (P <0.001). The association between lifetime smoking exposure and mean probing depth was statistically significant in water pipe as well as cigarette smokers controlling for age (P <0.001). Using multivariate analysis, besides smoking, the gingival and plaque indexes were associated with increased probing depth. The prevalence of periodontal disease defined as a minimum of 10 sites with a probing depth > or =5 mm was 19.5% in the total population, 30% in water pipe smokers, 24% in cigarette smokers, and 8% in non-smokers. The prevalence was significantly greater in water pipe and cigarette smokers compared to non-smokers (P <0.001). The relative risk for periodontal disease increased by 5.1- and 3.8-fold in water pipe and cigarette smokers, respectively, compared to non-smokers (P <0.001 and P <0.05, respectively). CONCLUSIONS: An association was observed between water pipe smoking and periodontal disease manifestations in terms of probing depth measurements. The impact of water pipe smoking was of largely the same magnitude as that of cigarette smoking. PMID- 16274312 TI - Systemic low-dose doxycycline and alendronate administration and serum interleukin-1beta, osteocalcin, and C-reactive protein levels in rats. AB - BACKGROUND: The aim of the present study was to evaluate the effects of systemic administration of low-dose doxycycline and a bisphosphonate, alendronate, on serum levels of interleukin-1beta (IL-1beta), osteocalcin (OC), and C-reactive protein (CRP) in experimental periodontitis in rats. METHODS: Experimental periodontitis was induced by repeated injection of purified lipopolysaccharide (LPS) derived from Escherichia coli endotoxin. Forty-seven adult male Sprague Dawley rats were divided into five study groups and given LPS, LPS + doxycycline, LPS + alendronate, LPS + doxycycline + alendronate, and saline control. At the end of the 1-week protocol, blood samples were obtained, and the rats were sacrificed. Serum samples were analyzed for IL-1beta, OC, and CRP concentrations by enzyme-linked immunosorbent assay (ELISA). The jaws were defleshed, and alveolar bone loss was assessed morphometrically. Data were evaluated statistically by non-parametric tests. RESULTS: Morphometric measurements revealed significantly more bone loss in the LPS group compared to the saline control group (P <0.05). Alendronate revealed slight inhibition on alveolar bone loss either alone or in combination with doxycycline (alveolar bone loss: 0.41 mm in alendronate and combined drug treatment groups versus 0.45 mm in LPS and doxycycline groups). Significantly higher IL-1beta levels were observed with alendronate either alone or in combination with doxycycline than in the LPS group (P <0.05). Combined administration of doxycycline and alendronate showed significantly higher levels of OC than all of the other groups (P <0.01). Serum CRP levels did not exhibit significant differences between the study groups. CONCLUSIONS: Alendronate either alone or in combination with doxycycline provided slight inhibition on LPS-induced alveolar bone resorption. The significantly increased serum OC level observed in the combined drug treatment group suggests that combined administration of alendronate and doxycycline might increase bone remodeling and thereby inhibit the progression of alveolar bone resorption in rats. PMID- 16274313 TI - Analysis of eosinophilic round bodies formed after injection of enamel matrix derivative into the backs of rats. AB - BACKGROUND: Enamel matrix derivative (EMD) is used in dental clinics for the regeneration of alveolar bone. Its effects have not yet been clarified, although it induces eosinophilic round bodies (ERBs) and cartilage formation at the injection site. The objective of this experiment was to examine the histopathologic and biochemical properties of ERBs formed after EMD injection. METHODS: The backs of Sprague-Dawley rats injected with various concentrations of EMD were examined histopathologically. For biochemical examinations, ERBs were microdissected out from the sections. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), matrix-assisted laser-desorption ionization time-of flight (MALDI-TOF) mass spectrometry (MS), and database analysis of ERBs were carried out. RESULTS: The histopathological findings were consistent with a foreign body reaction. Numerous ERBs were observed 7 days after injection of 30.0 mg/ml EMD. Histopathologically, ERBs did not contain polysaccharide, amyloid, or hemosiderin. The cells surrounding ERBs were not macrophages or vascular endothelial cells. SDS-PAGE of the microdissected ERBs revealed an intense band at around the 40-kDa region. MALDI-TOF MS showed that the spectrum for ERBs has only a single strong ion intensity. Analysis of the amino acid sequence revealed that the ERBs were composed of various molecular fragments, which all contained an identical seven amino acid sequence. In addition, these peptides are a component of amelogenin. CONCLUSIONS: A high concentration of EMD induces ERBs that consist of a 40-kDa protein which includes a constituent part of amelogenin. The ERBs (or remaining EMD) might promote mesenchymal cell differentiation into hard tissue-forming cells around the EMD injection site. PMID- 16274314 TI - In vitro, clinical, and microbiological evaluation of a linear oscillating device for scaling and root planing. AB - BACKGROUND: The purpose of this study was to conduct an in vitro and short-term clinical and microbiological evaluation of a linear oscillating device for scaling and root planing (SRP). A comparison was made between conventional ultrasonic scaling (US) and hand scaling (HS) with and without chlorhexidine. METHODS: In vitro, SRP was carried out on human teeth with calculus. Roots and cross-sections thereof were microscopically examined for the efficacy of calculus removal, hard tissue loss, and surface smoothness. In vivo, 11 patients with chronic periodontitis and single-rooted teeth in all quadrants with probing depths of > or =5 mm were selected. One quadrant was treated with linear oscillation and compared to US with chlorhexidine irrigation in the contralateral site. The other arch was treated with HS and compared to HS followed by laser disinfection. One hundred twenty teeth were assessed for clinical attachment level, probing depth, bleeding on probing, and suppuration at baseline and 7, 28, 90, and 180 days. Microbiologically, total numbers of bacteria and six specific periodontal pathogens were determined by quantitative polymerase chain reaction prior to and 1 and 28 days after SRP. Clinical and microbiological data were analyzed statistically with respect to the SRP method, patient specificity, and time effect. RESULTS: In vitro, linear oscillation preserved more root tissues but left more calculus (P <0.05). Significant improvements of all clinical and microbiological parameters were observed for all groups. However, 21 out of 24 tests demonstrated that the clinical microbiological correlations between linear oscillation and control groups did not differ (P <0.05). CONCLUSION: Linear oscillation scaling was clinically acceptable and microbiologically comparable to the control groups despite microscopic remnants of calculus observed in vitro. PMID- 16274315 TI - The expression profile of lipopolysaccharide-binding protein, membrane-bound CD14, and toll-like receptors 2 and 4 in chronic periodontitis. AB - BACKGROUND: This study aimed to investigate the interrelationship of in vivo expression of lipopolysaccharide-binding protein (LBP) and membrane-bound CD14 (mCD14) in human gingival tissues as well as the coexpression of Toll-like receptors (TLR) 2 and 4 in association with periodontal conditions. METHODS: Gingival biopsies were collected from 43 subjects with chronic periodontitis, including periodontal pocket tissues (PoTs) and clinically healthy gingival tissues (HT-Ps), and from 15 periodontally healthy subjects as controls (HT-Cs). The expression of LBP, CD14, TLR 2, and TLR 4 was detected by immunohistochemistry and reverse transcription-polymerase chain reaction (RT PCR). RESULTS: LBP and mCD14 peptides were simultaneously detected in 91% of PoTs, 85% of HT-Ps, and 100% of HT-Cs. LBP and mCD14 mRNAs were simultaneously detected in 55% of PoTs, 55% of HT-Ps, and 75% of HT-Cs. The expression of LBP was confined to the gingival epithelium, whereas mCD14 was observed around the epithelium-connective tissue interface. A positive correlation existed between LBP and mCD14 peptides in both detection expression (r(s) = 0.608; P <0.001) and expression levels (r = 0.304; P <0.05) of these two molecules. In PoTs, TLR 2 was detected in both pocket epithelia and macrophage-like cells in connective tissues, whereas TLR 4 was predominantly detected in connective tissues. In HT-Ps and HT-Cs, a weak expression of TLR 2 was found in gingival epithelia, and no TLR 4 expression was detected. In PoTs, mCD14 was codetected on CD68-labeled macrophages in the underlying connective tissues of pocket epithelium as well as on CD1a-labeled dendritic cells in the pocket epithelium and connective tissues interface. No similar expression profile was detected in HT-Ps and HT-Cs. CONCLUSIONS: This study suggests that the in vivo expression of LBP and mCD14 may be interrelated. Altered cellular expression profiles of mCD14 and TLR 2 and 4 in periodontal pocket tissues imply that these pattern recognition receptors may play a role in periodontal pathogenesis. PMID- 16274316 TI - CD38 expression in neutrophils from patients with localized aggressive periodontitis. AB - BACKGROUND: Localized aggressive periodontitis (LAgP) is associated with neutrophil dysfunction including decreased chemotaxis and reduced calcium entry. It has been suggested that CD38 is involved in chemotaxis. Little is known, however, about the relationship of CD38 and LAgP patients. In this study, we focused on the level of CD38 expression between LAgP and normal subjects and examined the involvement of CD38 in abnormal neutrophil chemotaxis of LAgP patients. METHODS: Neutrophils from 12 normal subjects and 12 LAgP patients were isolated from peripheral venous blood. Membrane associated proteins were extracted from cells with or without N-formylmethionine leucyl-phenylalanine (fMLP) stimulation. CD38 expression was measured using Western blotting. Band density was measured using an imaging densitometer. RESULTS: There was no statistical difference between normal subjects and LAgP patients in resting CD38 expression (basal level). However, the fMLP-stimulated neutrophils exhibited a significant decrease of CD38 expression in LAgP subjects compared to normal subjects. The decrease of CD38 was positively correlated with the defect in chemotactic migration to fMLP. CONCLUSION: These data suggest that the lower expression of CD38 in neutrophils may be related to altered neutrophil function in LAgP. PMID- 16274317 TI - Alveolar bone loss and aging: a model for the study in mice. AB - BACKGROUND: Several animal studies have shown a positive correlation between aging and alveolar bone loss (ABL). The purpose of this study was to develop a model for the study of ABL in mice and aging. METHODS: Mucoperiosteal flap surgery (MFS) was performed on the buccal aspect of the left side of the mandible (BL) in 72 CF(1) Mus domesticus mice and divided into three groups as follows: males, breeding (non-virgin) females, and virgin females. The MFS was performed in 3-, 6-, 9-, and 12-month-old animals under anesthesia. The buccal aspects of right hemimandibles were used as controls (BR). Animals were sacrificed under anesthesia 21 days after surgery. Mandibles were removed, defleshed, stained with toluidine blue, and photographed in a microscope. The photographs were digitized, and ABL was measured as the exposed root surface area (mm(2)). Blinded measurements were performed using a computer-assisted image analysis system. RESULTS: In terms of alveolar bone loss, the BL (operated) area showed a significant difference (paired Student t test; P <0.001) when compared to the BR area in all three groups. Sex and breeding differences were not observed in this experiment. ABL in the left hemimandibles was significantly larger in 3- (mean: 0.70; 95% confidence interval [CI]: 0.59 to 0.80) and 12-month-old animals (mean: 0.58; 95% CI: 0.46 to 0.71) than in 6- (mean: 0.39; 95% CI: 0.33 to 0.46) and 9 month-old animals (mean: 0.42; 95% CI: 0.35 to 0.48); P <0.001. CONCLUSION: The results suggest that 3-month-old CF(1) mice, in a sex and breeding independent process, could be a useful model for provoked alveolar bone loss studies in aging. PMID- 16274318 TI - Immediate occlusal loading of double acid-etched surface titanium implants in 41 consecutive full-arch cases in the mandible and maxilla: 6- to 74-month results. AB - BACKGROUND: The high success rate of dental implants has changed the quality of life for many patients. Immediate loading finds its application in some clinical cases and certainly adds another modality of treatment for the implant patient. Starting with a few implants immediately loaded with a bar overdenture in the mandible, the concept of immediate loading evolved to loading multiple implants in both the maxilla and mandible. METHODS: In this investigation, 41 consecutive patients who needed full-arch restorations were treated with 343 double acid etched surface implants between May 1998 and March 2004. The sample included smokers and bruxers. Twenty-three mandibular and 26 maxillary cases were treated, loading the implants within 48 hours, by using resin provisional prostheses, metal-reinforced provisional prostheses, or definitive prostheses (metal-acrylic or metal-ceramic). All implants were followed for 12 to 74 months. Follow-up consisted of clinical as well as radiographic examination. Furthermore, resonance frequency analysis was done in most of the implants during the last 2 years. RESULTS: The success rate obtained was 99.42% (only two mandibular implants that are still surviving were considered failures). The bone level was measured every year. The average radiographic bone level change was 0.56 mm at 12, 0.76 mm at 24, 0.84 mm at 36, 0.82 mm at 48, 0.83 mm at 60, and 0.94 mm at 72 months. CONCLUSION: A high success rate can be achieved when double acid-etched surface implants are immediately loaded with fixed full-arch restorations in the maxilla and mandible. PMID- 16274319 TI - Management of gingival recession by the use of an acellular dermal graft material: a 12-case series. AB - BACKGROUND: Different soft tissue defects can be treated by a variety of surgical procedures. Most of these techniques require the palatal area as a donor site. Recently, an acellular dermal graft has become available that can substitute for palatal donor tissue. METHODS: This study describes the surgical technique for gingival augmentation and root coverage and the results of 12 clinical cases. A comparison between the three most popular mucogingival procedures for root coverage is also presented. RESULTS: The results of the 12 patients and the 26 denuded surfaces have shown that we can obtain a mean root coverage of 74% with the acellular dermal graft. Thirteen out of the 26 denuded surfaces had complete root coverage. The average increase in keratinized tissue was 1.19 mm. It seems that the long-term results of the cases are stable. CONCLUSION: The proposed technique of root coverage with an acellular dermal graft can be a good alternative to soft tissue grafts for root coverage, and it should be part of our periodontal plastic surgery armamentarium. PMID- 16274320 TI - Periodontal microcirculatory abnormalities in patients with systemic sclerosis. AB - BACKGROUND: The aim of this study was to observe the differences in periodontal microcirculation between healthy patients and patients with systemic sclerosis (SSc). METHODS: Fifteen healthy patients and 15 patients with SSc were examined. Periodontal capillaroscopy was used to investigate the characteristics of microcirculation. The visibility, course, tortuosity, possible presence of microhemorrhages, average caliber of the capillary loops, and number of visible capillary loops per square millimeter were evaluated for each patient. RESULTS: The investigation was simple, non-invasive, and repeatable for each patient. In patients with SSc, it was possible to observe a reduced number of capillaries and a greater capillary diameter and tortuosity. CONCLUSIONS: Capillary alterations in patients with SSc are not limited to the nailfold bed but also occur in periodontal mucosa microcirculation. Such evidence could be extremely important in the pathogenesis and treatment of periodontal diseases in patients with SSc. PMID- 16274321 TI - Papillon-Lefevre syndrome: a successful outcome. AB - BACKGROUND: Papillon-Lefevre syndrome (PLS) is a rare autosomal recessive condition manifested clinically by hyperkeratosis of the palms and soles and rapidly progressive periodontitis resulting in loss of deciduous and permanent teeth. This case report describes the clinical periodontal findings and treatment of a 10-year-old male patient with PLS. The patient provided informed consent, and the study was conducted in accordance with the Helsinki Declaration of 1975, as revised in 2000. METHODS: Upon initial presentation, a full periodontal examination was completed. Conventional probing depths, clinical attachment levels (CAL), gingival index (GI), and plaque index (PI) were measured prior to initial therapy, which involved oral hygiene instruction and scaling and root planing. At reevaluation, initial treatment proved unsuccessful, and a surgical approach with concomitant systemic antibiotic therapy was implemented. In addition, the patient's dermatologist treated his palmoplantar keratoderma with systemic retinoids. Subsequently, the patient was placed on a strict 3-month maintenance protocol and was evaluated over a period of 1 year. RESULTS: Initial treatment with mechanical therapy, oral hygiene instruction, frequent recalls, and systemic antibiotics did not yield efficacious results. However, with the addition of surgical treatment, a favorable clinical outcome was obtained. CONCLUSIONS: Numerous treatment regimens for the periodontal disease seen in PLS can be found in the literature. We demonstrate successful treatment of the periodontal disease seen in this condition using mechanical therapy, systemic antibiotics, and surgical modalities; over a period of 1 year, we were able to achieve significant reductions in gingival inflammation and erythema. PMID- 16274322 TI - Progressive, generalized, apical idiopathic root resorption and hypercementosis. AB - BACKGROUND: Root resorption is a multifactorial process that results in loss of tooth structure. The causes of root resorption may vary, leading to several types of resorptions. Some factors have been identified and may be categorized into physiological resorption, local factors, systemic conditions, and idiopathic resorptions. The objective of this report was to present a case of a 17-year-old white female with progressive, generalized, apical idiopathic root resorption followed up for 34 months. METHODS: Two panoramic radiographs, 14 and 34 months after initial clinical and radiological examinations, showed the rapid progression of apical root resorption. Two molars, teeth #15 and #16, which had to be extracted, and a bone sample from the distal aspect of tooth #15 were processed for histologic analysis. RESULTS: Two millimeters apical to the cemento enamel junction, an abrupt increase in the cementum thickness was noted, amounting to 300 and 800 microm in teeth #15 and #16, respectively. The thickening of the cementum layer was due to an accelerated deposition of cellular intrinsic fiber cementum. An unusually high number of mineralization foci were observed in association with acellular extrinsic fiber cementum, and both free and fused cementicles were seen. In contrast to tooth #16, tooth #15 revealed extensive dentin replacement by a bone-like and a cementum-like tissue. Furthermore, ankylosis was demonstrated in tooth #15 and confirmed in the bone sample. CONCLUSIONS: At present, there is no preventive or therapeutic regimen for the type of root resorption seen in this case report. Treatment usually consists of the extraction of teeth with advanced lesions. PMID- 16274323 TI - President's address. PMID- 16274324 TI - Position paper: The role of supra- and subgingival irrigation in the treatment of periodontal diseases. AB - This position paper addresses the role of supra- and subgingival irrigation in the treatment of periodontal diseases. It was prepared by the Committee on Research, Science and Therapy of the American Academy of Periodontology. The document is divided into two portions, consisting of supragingival irrigation and subgingival irrigation. In their respective segments, these treatment techniques are assessed as monotherapies and as adjuncts to conventional treatment. The conclusions drawn in this paper represent the position of the American Academy of Periodontology regarding irrigation therapy in the treatment of periodontal diseases. PMID- 16274326 TI - Beta-blockers in post-traumatic stress disorder: uses and controversies. PMID- 16274328 TI - Selegiline orally disintegrating tablets for the treatment of Parkinson's disease. AB - Selegiline orally disintegrating tablet is a potent, selective and irreversible inhibitor of monoamine oxidase type B. It is delivered in an innovative formulation that largely bypasses the gut and first-pass hepatic metabolism by allowing transbuccal absorption. Selegiline orally disintegrating tablet dissolves in the mouth within seconds and is rapidly absorbed directly into the systemic circulation, increasing parent drug bioavailability and lowering plasma metabolites compared with conventional oral formulations. Adding selegiline orally disintegrating tablet to levodopa in patients experiencing 'wearing-off' episodes significantly decreases off time and increases dyskinesia-free 'on' time. Adding selegiline orally disintegrating tablet to levodopa also significantly improves Unified Parkinson's Disease Rating Scale motor scores and patients' and physicians' ratings of disease severity. Selegiline orally disintegrating tablet has been demonstrated to be safe and well tolerated in placebo-controlled clinical trials. PMID- 16274329 TI - Risperidone in pervasive developmental disorders. AB - Atypical antipsychotics are increasingly used in the treatment of severe behavioral disturbances in people with pervasive developmental disorders. Among these compounds, risperidone has been the most broadly studied. This drug profile will review the available data on the use of risperidone in individuals with pervasive developmental disorders. In addition, the chemistry, pharmacokinetics, pharmacodynamics and metabolism of risperidone are discussed. Emphasis will be placed on adverse effects associated with risperidone in these people, including a discussion of measures that can be taken to monitor for these effects. Finally, a discussion of the future direction of risperidone use in this population and key issues will be presented. PMID- 16274330 TI - Cladribine for multiple sclerosis: review and current status. AB - In the 1990s, cladribine was developed as an adenosine deaminase-resistant nucleoside analog with selective lymphotoxic specificity in the hope that it might become useful in the treatment of some lymphoid neoplasms and autoimmune disorders. Several clinical trials demonstrated very significant effectiveness and safety of cladribine in the cure of hairy-cell leukemia, and the control of many other lymphoid malignancies. Cladribine was also extensively tested in selected autoimmune disorders, most notably in multiple sclerosis, with evidence of efficacy, tolerability and acceptable side effects/toxicity. The previous clinical studies and current status of cladribine for the treatment of multiple sclerosis are considered in this drug profile. In January 2005, Serono and IVAX announced plans to initiate a Phase III study of a specially formulated oral tablet of cladribine (Mylinax, Serono and IVAX) for the treatment of relapsing forms of multiple sclerosis. The proposed study will be the first large multicenter randomized controlled clinical trial of oral cladribine in multiple sclerosis. PMID- 16274332 TI - Psychologic treatment of patients with psychogenic nonepileptic seizures. AB - Psychogenic nonepileptic seizures are relatively common, often disabling and costly to patients and society. Most authorities consider psychologic treatment as the therapeutic intervention of choice. This review is intended primarily for psychologists and therapists who treat patients with psychogenic nonepileptic seizures, and for neurologists who make the diagnosis and wish to find out more about psychologic treatment options. The first section describes the nature and etiology of psychogenic nonepileptic seizures. General questions regarding the psychologic treatment of patients with psychogenic nonepileptic seizures are addressed, before discussing specific therapeutic approaches. The final part summarizes the authors' views on optimal treatment and the direction of future research. PMID- 16274331 TI - Lamotrigine for neuropathic pain. AB - Lamotrigine is an antiepileptic drug that stabilizes neural membranes by blocking the activation of voltage-sensitive sodium channels and inhibiting the presynaptic release of glutamate. Full length reports of five open trials and six out of seven randomized controlled trials (plus two abstracts) have demonstrated the efficacy of lamotrigine in the treatment of various forms of neuropathic pain. The present drug profile provides a review of the pharmacologic properties of lamotrigine, the clinical evidence related to its efficacy and safety, and discusses the current and future role of the drug in the treatment of neuropathic pain. PMID- 16274333 TI - Idiopathic generalized epilepsies: clinical and electroencephalogram diagnosis and treatment. AB - This review concentrates on the principles of the clinical and electroencephalogram diagnosis of idiopathic generalized epilepsies and their treatment. The electroclinical variability of the main seizure types is detailed and particular emphasis is placed on the differential diagnosis from other seizures and nonepileptic conditions that is essential for the optimal management of these patients. The authors review the various idiopathic generalized epilepsy subsyndromes and conditions that are included in both the 1989 International League Against Epilepsy classification system and the recently proposed International League Against Epilepsy scheme, but also syndromes and forms that have not been formally recognized. Finally, the authors describe the principles of antiepileptic drug treatment with the old and newer drugs, and their specific indications and contraindications in the various syndromes and seizure types. PMID- 16274334 TI - Use of the ketogenic diet as a treatment for epilepsy refractory to drug treatment. AB - The ketogenic diet is a high-fat, low-carbohydrate and low-protein diet used in the treatment of epilepsy that does not respond to antiepileptic drugs. The diet has been found to be very effective in treating intractable epilepsy in children. There is also some evidence that the diet is useful in treating drug-resistant epilepsy in infants, adolescents and adults. This paper traces the history and development of the ketogenic diet and reviews the clinical and animal research investigating its effects. PMID- 16274335 TI - Pharmacologic management of convulsive status epilepticus in childhood. AB - The incidence of convulsive status epilepticus in children is approximately 20 50/100,000/year, and is an emergency requiring prompt medical intervention. Prolonged seizures lasting over 5 min are unlikely to stop spontaneously, and time-to-treatment influences treatment response. Prolonged seizures should thus be treated as early status epilepticus. Mortality and morbidity increase significantly with the length of ongoing seizure activity, especially after 60 min. Benzodiazepines remain the first-line drug therapy due to their rapid onset of action. Recent studies imply that buccal midazolam is more effective and easier to administer than rectal diazepam. Phenytoin/fosphenytoin and phenobarbital administered intravenously remain the second-line treatments of choice, whilst barbiturates and midazolam as intravenous anesthetics are used for third-line treatment. Electroencephalogram monitoring is essential to evaluate the electrophysiologic treatment response and depth of anesthesia, especially in refractory status epilepticus. In the future, more individualized protocols and pathways are needed in order to optimize treatment responses. Randomized clinical trials are needed to evaluate new treatment protocols, which should not only stop the seizures more effectively but also be safer and include some neuroprotective elements to halt the cascade of neuronal injury and minimize the risk for neurologic morbidity caused by the convulsive status epilepticus. PMID- 16274336 TI - Treatment options and paradigms in childhood temporal lobe epilepsy. AB - Temporal lobe epilepsy in adults is a relatively homogenous syndrome with hippocampal sclerosis being its most common pathologic substrate. In the pediatric age group, low-grade neoplasms and cortical dysplasia are much more common than hippocampal sclerosis. Pediatric temporal lobe epilepsy has distinct semiologic, electrophysiologic and imaging characteristics as compared with its adult counterpart. The various treatment options for pediatric temporal lobe epilepsy include antiepileptic drugs, resective surgery, vagal nerve stimulation and the ketogenic diet. In spite of the multiple antiepileptic drugs currently available, 5-10% of all newly diagnosed cases will remain intractable to medical therapy and should be referred for presurgical evaluation. Resective surgery offers the best chance of seizure freedom in carefully selected patients. Future areas of research include new drug development, better imaging and localization techniques, and brain stimulation. PMID- 16274337 TI - A matter of motion or an emotional matter? Management of depression in Parkinson's disease. AB - Depression is one of the most frequent comorbidities occurring in Parkinson's disease, affecting up to 50% of patients. Depression is associated with severe negative symptoms and has been shown to contribute to an increased rate of decline of both cognitive and motor function, profoundly impacting on the patient's quality of life. The symptoms of depression overlap with the motor features of Parkinson's disease, making detection difficult. Moreover, the lack of specialized screening tools means that depression remains undiagnosed and untreated in a high percentage of patients. However, depression in Parkinson's disease, when identified early, can be effectively treated with a variety of antidepressant medications, improving quality of life and preserving daily function. The focus of this review is to provide an overview of current knowledge regarding depression in Parkinson's disease, followed by a practical discussion addressing the issues of the detection, diagnosis and treatment. PMID- 16274338 TI - Recent advances in Parkinson's disease therapy: use of monoamine oxidase inhibitors. AB - Monoamine oxidase inhibitors inhibit dopamine metabolism and are therefore effective in treating Parkinson's disease, a condition associated with progressive striatal dopamine deficiency secondary to degeneration of dopaminergic neurons in the substantia nigra. Selegiline is currently the most widely used monoamine oxidase-B inhibitor for Parkinson's disease, but has a low and variable bioavailability, and is metabolized to L-methamphetamine and L amphetamine that carry a risk for potential neurotoxicity. There are two new approaches that circumvent these potential disadvantages. First, selegiline orally disintegrating tablets provide a novel delivery form of selegiline, avoiding first pass metabolism by rapid absorption through the oral mucosa, thus leading to significantly lower plasma concentrations of L-metamphetamine and L amphetamine. Selegiline orally disintegrating tablets prove to be clinically effective and safe in patients with moderately advanced Parkinson's disease. Second, rasagiline is a new monoamine oxidase inhibitor, without known neurotoxic metabolites. In large clinical trials, rasagiline proves effective as monotherapy in early Parkinson's disease, as well as adjunctive therapy to levodopa in advanced disease. Clinical data suggest, in addition, a disease-modifying effect of rasagiline that may correlate with neuroprotective activity of monoamine oxidase-B inhibitors in animal models of Parkinson's disease. PMID- 16274339 TI - Combination pharmacotherapy for neuropathic pain: current evidence and future directions. AB - Current drugs reduce neuropathic pain and improve mood and quality of life. However, as single agents they are limited by incomplete efficacy and dose limiting adverse effects. Recent experimental and clinical data support the potential of combination pharmacotherapy for neuropathic pain. Therapeutic benefits may include greater efficacy, lower doses and fewer adverse effects. Due to potential adverse, as well as beneficial, drug interactions, safety and efficacy of specific combinations must be empirically evaluated. Techniques such as isobolographic analysis, response-surface modeling and other model-free tests have been used in order to characterize analgesic interactions as antagonistic, additive or synergistic. Whether synergistic or not, a clinically useful combination could simply have additive or even subadditive analgesia, provided that there is less additivity for side effects. Despite widespread clinical use, there are surprisingly few published observations on combination therapy for neuropathic pain. This review discusses future directions and proposes research strategies aimed at bridging current knowledge gaps, including safety, compliance and cost-effectiveness; discovering optimal drug combinations and dose ratios; comparing concurrent with sequential combination therapy; and combining more than two drugs. Continued close integration of basic and clinical sciences is crucial in further harnessing the potential of combination pharmacotherapy in neuropathic pain. PMID- 16274340 TI - New therapeutics in spine metastases. AB - The number of patients who will develop metastatic spinal tumors is estimated to be between 5 and 10% of all cancer patients. As the therapy for systemic cancer improves, the number of patients developing symptomatic spinal tumors that require local therapy will increase. Over the last 10 years there has been a dramatic evolution in our ability to treat spinal tumors. These advances have not only been created by improvements in surgical techniques and instrumentation, but also developments in radiographic imaging, radiation therapy and chemotherapy. It is important for spine surgeons, radiologists, and radiation and medical oncologists to continue developing techniques for spinal salvage that will improve pain relief, achieve mechanical stability, improve or maintain neurologic function and sustain local tumor control. The evolution of these technologies will help to provide palliation and improve quality of life for patients with metastatic disease. PMID- 16274342 TI - Asthma in Australia 2005. AB - A recent report outlines the good and the bad news about asthma. PMID- 16274341 TI - More students and less patients: the squeeze on medical teaching resources. AB - We urgently need to expand clinical teaching into the private sector. PMID- 16274343 TI - Systemic adjuvant therapies for early breast cancer: 15-year results for recurrence and survival. PMID- 16274344 TI - Cost-effectiveness of rehabilitation after an acute coronary event: a randomised controlled trial. AB - OBJECTIVE: To estimate the incremental effects on cost and quality of life of cardiac rehabilitation after an acute coronary syndrome. DESIGN: Open randomised controlled trial with 1 year's follow-up. Analysis was on an intention-to-treat basis. SETTING: Two tertiary hospitals in Sydney. INTERVENTION: 18 sessions of comprehensive exercise-based outpatient cardiac rehabilitation or conventional care as provided by the treating doctor. PARTICIPANTS: 113 patients aged 41-75 years who were self-caring and literate in English. Patients with uncompensated heart failure, uncontrolled arrhythmias, severe and symptomatic aortic stenosis or physical impairment were excluded. MAIN OUTCOME MEASURES: Costs (hospitalisations, medication use, outpatient visits, investigations, and personal expenses); and measures of quality of life. Incremental cost per quality adjusted life year (QALY) saved at 1 year (this estimate combines within-study utility effects with reported 1-year risk of survival and treatment effects of rehabilitation on mortality). Sensitivity analyses around a base case estimate included alternative assumptions of no treatment effect on survival, 3 years of treatment effect on survival and variations in utility. RESULTS: The estimated incremental cost per QALY saved for rehabilitation relative to standard care was 42,535 US dollars when modelling included the reported treatment effect on survival. This increased to 70,580 US dollars per QALY saved if treatment effect on survival was not included. The results were sensitive to variations in utility and ranged from 19,685 US dollars per QALY saved to rehabilitation not being cost effective. CONCLUSIONS: The effects on quality of life tend to reinforce treatment advantages on survival for patients having postdischarge rehabilitation after an acute coronary syndrome. The estimated base case incremental cost per QALY saved is consistent with those historically accepted by decision making authorities such as the Pharmaceutical Benefits Advisory Committee. PMID- 16274345 TI - Barriers to delivering asthma care: a qualitative study of general practitioners. AB - OBJECTIVES: To ascertain what general practitioners' priorities are for achieving optimal outcomes in people with asthma, and the barriers they face in delivering this care. DESIGN: A qualitative study using the Nominal Group Technique (a highly structured meeting to gain information from experts about a particular issue) was conducted between August 2002 and September 2003. GPs in six discussion groups were asked "What do you think is needed to achieve best outcomes for asthma care?" To augment analysis of the discussion, sessions were taped and transcribed. PARTICIPANTS: Forty-nine GPs were recruited: 34 from metropolitan and 15 from rural areas. RESULTS: All groups nominated asthma education for patients and continuing professional education for GPs as major priorities, but they also described educational and structural barriers to achieving these priorities. Other priorities were: medication adherence, facilitating regular patient review, negotiated treatment/management plans, making the correct diagnosis, increased remuneration and consultation time, and safer asthma medications and access to these. Health promotion initiatives and increased public awareness were also priorities. Spirometry was a significant cause of uncertainty. Overall, written asthma action plans were not considered a high priority. CONCLUSIONS: Remarkable consistency was found between GPs' priorities for delivering best asthma care. Our study identified barriers to asthma guideline adherence, including accessible, relevant education for GPs, and structural, time and cost barriers GPs must overcome in providing asthma treatment and patient education. PMID- 16274346 TI - Barriers to student access to patients in a group of teaching hospitals. AB - OBJECTIVES: To determine the number of patients in our teaching hospitals who were, on any given day, both available and willing to see medical students. DESIGN AND SETTING: Repeated cross-sectional audit in four teaching hospitals in the greater Newcastle area of New South Wales (one tertiary referral hospital, two district general hospitals, and one hospital combining general medicine and surgery with specialised oncology services). Audits were conducted three times, 2 months apart. PARTICIPANTS: All adult inpatients in the four hospitals. MAIN OUTCOME MEASURES: Numbers of patients present and accessible to students, present but inaccessible, absent, or unfit to be seen for clinical reasons; numbers of patients who agreed to history-taking and physical examination by a medical student. RESULTS: Of 1960 patients, 959 (49%) were present and accessible to students. Only 11% were absent, and the most common reason students could not see patients was that the patients were said by nursing staff to be unfit to see medical students (25%). Of those present and accessible, 70% said they would agree to provide a history, and 67% that they would agree to physical examination. CONCLUSIONS: Across all four teaching hospitals about 200-250 patients are available and willing to see medical students on any given day. This is too few to provide our current student population of 500 with extensive clinical experience. PMID- 16274348 TI - Adult domiciliary oxygen therapy: a patient's perspective. PMID- 16274347 TI - A systematic review and economic analysis of drug-eluting coronary stents available in Australia. AB - OBJECTIVES: To compare the safety, effectiveness and cost-effectiveness of drug eluting coronary stents used in Australia with bare-metal stents and determine whether the benefits are greater for high-risk subgroups. DATA SOURCES: MEDLINE, Pre-Medline, EMBASE, Current Contents, CINAHL and the Cochrane Library database were searched to identify eligible randomised controlled trials and systematic reviews published in English between January 1966 and June 2004. STUDY SELECTION: Seven randomised controlled trials that assessed polymer-based paclitaxel- or sirolimus-eluting stents versus bare-metal stents in patients with coronary atherosclerosis and reported on stent thrombosis, mortality, myocardial infarction, coronary artery bypass grafting or target lesion revascularisation. DATA EXTRACTION: Two independent reviewers appraised eligible studies and extracted data. Relative risks (RRs) were calculated for each outcome and pooled using the Mantel-Haenszel method. DATA SYNTHESIS: Rates of stent thrombosis, mortality, myocardial infarction and bypass grafts did not differ by stent type. Drug-eluting stents (DESs) resulted in a 71%-80% lower risk of revascularisation at 12 months (RR 0.29 [95% CI, 0.20-0.43] for paclitaxel-eluting stents [n = 1593 patients]; RR 0.20 [95% CI, 0.13-0.29] for sirolimus-eluting stents [n = 1296 patients]). Similar benefits were seen in several high-risk subgroups of patients: those with diabetes, lesion length > 20 mm and target-vessel diameter < or = 2.5 mm. The benefits of DESs in these high-risk groups over lower-risk groups were inconclusive because of low numbers. The cost per revascularisation avoided by using DESs was 3,750-6,100 Australian dollars, with an estimated cost per quality-adjusted-life-year (QALY) gained of 46,829-76,467 Australian dollars. In sensitivity analyses, estimates varied from DESs being cost-saving to costing an additional 314,385 Australian dollars per QALY gained. CONCLUSIONS: DESs are effective in reducing revascularisation. Estimates of cost-effectiveness are very sensitive to changes in estimates of their true effects in clinical practice, market price and the number of stents used per patient. Decisions to limit DESs to only patients at the highest risk of restenosis may improve their cost effectiveness but will need to be reassessed when evidence is available to compare absolute benefits between patient groups. PMID- 16274351 TI - Tubulointerstitial nephritis and uveitis syndrome: sore eyes and sick kidneys. PMID- 16274350 TI - Teaching on the run tips 11: the junior doctor in difficulty. PMID- 16274352 TI - Telling you our story: how apology and action relate to health and social problems in Aboriginal and Torres Strait Islander communities. PMID- 16274353 TI - The use of diagnostic imaging in sports medicine. AB - Imaging should only be undertaken if it is likely to influence patient management. The dose of ionising radiation to the patient should be considered. Requesting the appropriate imaging method requires an understanding of the pathological process. Plain x-ray should still generally be the first imaging technique; exceptions include some forms of superficial tendinopathy, in which ultrasound may be more appropriate, and situations where radiation exposure is contraindicated, such as in a pregnant patient. The cost of the examination to the patient and the community should also be considered (eg, ultrasound v magnetic resonance imaging). PMID- 16274354 TI - Two linked cases of legionellosis with an unusual industrial source. PMID- 16274355 TI - Hepatitis E virus: overseas epidemics and Victorian travellers. PMID- 16274356 TI - Sight-seeing in the Solomon Islands. PMID- 16274357 TI - Health development assistance works: a Pacific example. PMID- 16274358 TI - Vision loss in Australia. PMID- 16274359 TI - "GP Psych Opinion": evaluation of a psychiatric consultation service. PMID- 16274360 TI - Adult domiciliary oxygen therapy. Position statement of the Thoracic Society of Australia and New Zealand. PMID- 16274361 TI - Spectroscopic and kinetic characterization of the light-dependent enzyme protochlorophyllide oxidoreductase (POR) using monovinyl and divinyl substrates. AB - The enzyme POR [Pchlide (protochlorophyllide) oxidoreductase] catalyses the reduction of Pchlide to chlorophyllide, which is a key step in the chlorophyll biosynthesis pathway. This light-dependent reaction has previously been studied in great detail but recent reports suggest that a mixture of MV (monovinyl) and DV (divinyl) Pchlides may have influenced some of these properties of the reaction. Low-temperature absorbance and fluorescence spectroscopy have revealed several spectral differences between MV and DV Pchlides, which were purified from a Rhodobacter capsulatus strain that was shown to contain a mixture of the two pigments. A thorough steady-state kinetic characterization using both Pchlide forms demonstrates that neither pigment appears to affect the kinetic properties of the enzyme. The reaction has also been monitored following illumination at low temperatures and was shown to consist of an initial photochemical step followed by four 'dark' steps for both pigments. However, minor differences were observed in the spectral properties of some of the intermediates, although the temperature dependency of each step was nearly identical for the two pigments. This work provides the first detailed kinetic and spectroscopic study of this unique enzyme using biologically important MV and DV substrate analogues. It also has significant implications for the DV reductase enzyme, which is responsible for converting DV pigments into their MV counterparts, and its position in the sequence of reactions that comprise the chlorophyll biosynthesis pathway. PMID- 16274362 TI - Human acyl-CoA:cholesterol acyltransferase 2 gene expression in intestinal Caco-2 cells and in hepatocellular carcinoma. AB - Humans express two ACAT (acyl-CoA:cholesterol acyltransferase) genes, ACAT1 and ACAT2. ACAT1 is ubiquitously expressed, whereas ACAT2 is primarily expressed in intestinal mucosa and plays an important role in intestinal cholesterol absorption. To investigate the molecular mechanism(s) responsible for the tissue specific expression of ACAT2, we identified five cis-elements within the human ACAT2 promoter, four for the intestinal-specific transcription factor CDX2 (caudal type homeobox transcription factor 2), and one for the transcription factor HNF1alpha (hepatocyte nuclear factor 1alpha). Results of luciferase reporter and electrophoretic mobility shift assays show that CDX2 and HNF1alpha exert a synergistic effect, enhancing the ACAT2 promoter activity through binding to these cis-elements. In undifferentiated Caco-2 cells, the ACAT2 expression is increased when exogenous CDX2 and/or HNF1alpha are expressed by co-transfection. In differentiated Caco-2 cells, the ACAT2 expression significantly decreases when the endogenous CDX2 or HNF1alpha expression is suppressed by using RNAi (RNA interference) technology. The expression levels of CDX2, HNF1alpha, and ACAT2 are all greatly increased when the Caco-2 cells differentiate to become intestinal like cells. These results provide a molecular mechanism for the tissue-specific expression of ACAT2 in intestine. In normal adult human liver, CDX2 expression is not detectable and the ACAT2 expression is very low. In the hepatoma cell line HepG2 the CDX2 expression is elevated, accounting for its elevated ACAT2 expression. A high percentage (seven of fourteen) of liver samples from patients affected with hepatocellular carcinoma exhibited elevated ACAT2 expression. Thus, the elevated ACAT2 expression may serve as a new biomarker for certain form(s) of hepatocellular carcinoma. PMID- 16274363 TI - Agonist-evoked inositol trisphosphate receptor (IP3R) clustering is not dependent on changes in the structure of the endoplasmic reticulum. AB - The size and number of IP3R (inositol 1,4,5-trisphosphate receptor) clusters located on the surface of the ER (endoplasmic reticulum) is hypothesized to regulate the propagation of Ca2+ waves in cells, but the mechanisms by which the receptors cluster are not understood. Using immunocytochemistry, live-cell imaging and heterologous expression of ER membrane proteins we have investigated IP3R clustering in the basophilic cell line RBL-2H3 following the activation of native cell-surface antigen receptors. IP3R clusters are present in resting cells, and upon receptor stimulation, form larger aggregates. Cluster formation and maintenance required the presence of extracellular Ca2+ in both resting and stimulated cells. Using transfection with a marker of the ER, we found that the ER itself also showed structural changes, leading to an increased number of 'hotspots', following antigen stimulation. Surprisingly, however, when we compared the ER hotspots and IP3R clusters, we found them to be distinct. Imaging of YFP (yellow fluorescent protein)-IP3R transfected in to living cells confirmed that IP3R clustering increased upon stimulation. Photobleaching experiments showed that the IP3R occupied a single contiguous ER compartment both before and after stimulation, suggesting a dynamic exchange of IP3R molecules between the clusters and the surrounding ER membrane. It also showed a decrease in the mobile fraction after cell activation, consistent with receptor anchoring within clusters. We conclude that IP3R clustering in RBL-2H3 cells is not simply a reflection of bulk-changes in ER structure, but rather is due to the receptor undergoing homotypic or heterotypic protein-protein interactions in response to agonist stimulation. PMID- 16274364 TI - Type 2 diabetes mellitus and risk of developing urinary incontinence. AB - OBJECTIVES: To evaluate the association between type 2 diabetes mellitus (DM) and development of urinary incontinence in women. DESIGN: Prospective, observational study. SETTING: The Nurses' Health Study cohort. PARTICIPANTS: Eighty-one thousand eight hundred forty-five women who reported information on urinary function in 1996. MEASUREMENTS: Self-reported, physician-diagnosed DM was ascertained using questionnaire from 1976 to 1996 and confirmed using standard criteria. Self-reported urinary incontinence, defined as leakage at least weekly, was ascertained in 1996 and 2000. Logistic regression models were used to calculate multivariate-adjusted relative risks (RRs) and 95% confidence intervals (CIs) for the relationship between DM (as of 1996) and prevalent and incident incontinence. RESULTS: The risk of prevalent incontinence (multivariate RR=1.28, 95% CI=1.18-1.39) and incident incontinence (multivariate RR=1.21, 95% CI=1.02 1.43) was significantly greater in women with DM than women without. Using a validated severity index, risk of developing severe incontinence was even more substantial in women with DM than in those without (multivariate RR=1.40, 95% CI=1.15-1.71 for leakage enough to wet the underwear; RR=1.97, 95% CI=1.24-3.12 for leakage enough to wet the outer clothing). In addition, risk of incontinence increased with duration of DM (P-trend=.03 for prevalent incontinence; P=.001 for incident incontinence). CONCLUSION: DM independently increases risk of urinary incontinence in women. Because risk of incontinence appeared associated with longer duration of DM, even delaying the onset of DM could have important public health implications. PMID- 16274365 TI - Mortality and adverse health events in newly admitted nursing home residents with and without dementia. AB - OBJECTIVES: To evaluate the association between dementia and mortality, adverse health events, and discharge disposition of newly admitted nursing home residents. It was hypothesized that residents with dementia would die at a higher rate and develop more adverse health events (e.g., infections, fevers, pressure ulcers, falls) than residents without dementia because of communication and self care difficulties. DESIGN: An expert clinician panel diagnosed an admission cohort from a stratified random sample of 59 Maryland nursing homes, between 1992 and 1995. The cohort was followed for up to 2 years or until discharge. SETTING: Fifty-nine Maryland nursing homes. PARTICIPANTS: Two thousand one hundred fifty three newly admitted residents aged 65 and older not having resided in a nursing home for 8 or more days in the previous year. MEASUREMENTS: Mortality, infection, fever, pressure ulcers, fractures, and discharge home. RESULTS: Residents with dementia had significantly lower overall rates of infection (relative risk (RR)=0.77, 95% confidence interval (CI)=0.70-0.85) and mortality (RR=0.61, 95% CI=0.53-0.71) than those without dementia, whereas rates of fever, pressure ulcers, and fractures were similar for the two groups. These results persisted when rates were adjusted for demographic characteristics, comorbid conditions, and functional status. During the first 90 days of the nursing home stay, residents with dementia had significantly lower rates of mortality if not admitted for rehabilitative care under a Medicare qualifying stay (RR=0.25, 95% CI=0.14-0.45), were less often discharged home (RR=0.33, 95% CI=0.28-0.38), and tended to have lower fever rates (RR=0.78, 95% CI=0.63-0.96) than residents without dementia. CONCLUSION: Newly admitted nursing home residents with dementia have a profile of health events that is distinct from that of residents without dementia, indicating that the two groups have different long-term care needs. Results suggest that further investigation of whether residents with dementia can be well managed in alternative residential settings would be valuable. PMID- 16274366 TI - Longitudinal analysis of the relationship between regular eye examinations and changes in visual and functional status. AB - OBJECTIVES: To determine whether regular eye examinations are associated with a greater or lesser rate of loss of ability to read newsprint, onset of blindness or low vision, or onset of limitations in instrumental activities of daily living (IADLs) and activities of daily living (ADLs). DESIGN: A sample of 14,215 Medicare beneficiaries observed between 1994 and 1999 linked to the 1994 and 1999 National Long-Term Care Surveys (NLTCS). Effects of annual examinations were assessed using instrumental variables. SETTING: The Medicare-linked NLTCS is representative of U.S. elderly persons from 1994 to 1999. PARTICIPANTS: Longitudinal observational study of persons aged 65 and older. MEASUREMENTS: Change in self-reported and provider-reported vision and change in functional limitations associated with vision related to the number of years with eye examinations and other factors. RESULTS: Persons with more-regular eye examinations between 1994 and 1998 were less likely to have experienced a decline in vision or in functional status between 1994 and 1999. On average, an additional year with an eye examination was associated with a decrease in the probability of becoming unable to read newsprint of 0.12 (P=.03), a lower probability of onset of low vision or blindness of 0.009 (P=.06), and a decrease in the probability that the number of functional limitations increased of 0.13 (P=.002) for IADLs and 0.05 (P=.003) for ADLs. CONCLUSION: Elderly persons who have regular eye examinations experience less decline in vision and functional status. PMID- 16274367 TI - Magnesium intake from food and supplements is associated with bone mineral density in healthy older white subjects. AB - OBJECTIVES: To determine whether magnesium intake from supplemental and dietary sources is associated with bone mineral density (BMD) in older men and women. DESIGN: Cross-sectional. SETTING: Memphis, Tennessee, and Pittsburgh, Pennsylvania. PARTICIPANTS: Two thousand thirty-eight older black and white men and women aged 70 to 79 at baseline enrolled in the Health, Aging and Body Composition Study. MEASUREMENTS: Dietary intake of magnesium was assessed using a semiquantitative food frequency questionnaire, and supplement data were collected based on a medication inventory. BMD of the whole body was obtained using a fan beam densitometer. Additional covariates included age, body mass index (BMI), smoking status, alcohol use, physical activity, estrogen use, and supplemental calcium (Ca) and vitamin D use. RESULTS: In white, but not black, men and women, magnesium intake was positively associated with BMD of the whole body after adjustment for age, self-report of osteoporosis or fracture in adulthood, caloric intake, Ca and vitamin D intake, BMI, smoking status, alcohol intake, physical activity, thiazide diuretic use, and estrogen use in women (P=.05 for men and P=.005 for women). BMD was 0.04 g/cm2 higher in white women and 0.02 g/cm2 higher in white men in the highest than in the lowest quintile of magnesium intake. CONCLUSION: Greater magnesium intake was significantly related to higher BMD in white women and men. The lack of association observed in black women and men may be related to differences in Ca regulation or in nutrient reporting. PMID- 16274368 TI - Should older people in residential care receive vitamin D to prevent falls? Results of a randomized trial. AB - OBJECTIVES: To determine whether vitamin D supplementation can reduce the incidence of falls and fractures in older people in residential care who are not classically vitamin D deficient. DESIGN: Randomized, placebo-controlled double blind, trial of 2 years' duration. SETTING: Multicenter study in 60 hostels (assisted living facilities) and 89 nursing homes across Australia. PARTICIPANTS: Six hundred twenty-five residents (mean age 83.4) with serum 25-hydroxyvitamin D levels between 25 and 90 nmol/L. INTERVENTION: Vitamin D supplementation (ergocalciferol, initially 10,000 IU given once weekly and then 1,000 IU daily) or placebo for 2 years. All subjects received 600 mg of elemental calcium daily as calcium carbonate. MEASUREMENTS: Falls and fractures recorded prospectively in study diaries by care staff. RESULTS: The vitamin D and placebo groups had similar baseline characteristics. In intention-to-treat analysis, the incident rate ratio for falling was 0.73 (95% confidence interval (CI)=0.57-0.95). The odds ratio for ever falling was 0.82 (95% CI=0.59-1.12) and for ever fracturing was 0.69 (95% CI=0.40-1.18). An a priori subgroup analysis of subjects who took at least half the prescribed capsules (n=540), demonstrated an incident rate ratio for falls of 0.63 (95% CI=0.48-0.82), an odds ratio (OR) for ever falling of 0.70 (95% CI=0.50-0.99), and an OR for ever fracturing of 0.68 (95% CI=0.38 1.22). CONCLUSION: Older people in residential care can reduce their incidence of falls if they take a vitamin D supplement for 2 years even if they are not initially classically vitamin D deficient. PMID- 16274369 TI - Physicians' assessments of adjuvant tamoxifen's effectiveness in older patients with primary breast cancer. AB - OBJECTIVES: To examine physicians' assessments of tamoxifen effectiveness in breast cancer patients, identify predictors of these assessments, and estimate the relationship between these assessments and receipt of tamoxifen prescription. DESIGN: A cohort of breast cancer patients aged 65 and older at diagnosis and their physicians were surveyed using mailed questionnaires and telephone interviews. SETTING: Community and academic hospitals in Rhode Island; North Carolina; Minnesota; and Los Angeles, California between 1996 and 1998. PARTICIPANTS: Physicians completed treatment recommendation forms for 496 of 865 Stage Ic to IIIa breast cancer patients. MEASUREMENTS: Visual scales measured physicians' assessments of the risk that individual patients would have a breast cancer recurrence or die of breast cancer with, and without, tamoxifen therapy. RESULTS: The mean risk ratio+/-standard deviation comparing risk of recurrence without tamoxifen with the risk with tamoxifen was 1.8+/-1.0 and for breast cancer mortality was 1.8+/-1.2. Only estrogen-receptor status and enrollment site emerged as significant predictors of recurrence and mortality risk ratios in regression models. Patients for whom the physician estimated that the recurrence or mortality risk doubled without tamoxifen were more likely to receive a tamoxifen prescription than patients for whom the physician estimated that tamoxifen would have no effect (odds ratio (OR)=1.4, 95% confidence interval (CI)=0.98-2.1 for recurrence risk, OR=1.8; 95% CI=1.2-2.6 for mortality risk). CONCLUSION: Estrogen receptor status most strongly influenced physicians' assessments of tamoxifen's effectiveness in individual patients; this effectiveness was not found to be associated with advancing patient age. Estrogen receptor status and enrollment site were related to receipt of tamoxifen prescription, but advancing age was not after accounting for physician's individualized assessment of tamoxifen's effectiveness. These findings suggest that an evidence-based approach for hormonal therapy has been widely adopted for care of older patients with breast cancer. PMID- 16274370 TI - Treatment for breast cancer in patients with Alzheimer's disease. AB - OBJECTIVES: To report use of breast cancer treatment (surgery, radiation, and chemotherapy) by patients with Alzheimer's disease (AD). DESIGN: Retrospective cohort study. SETTING: Surveillance, Epidemiology, and End Results (SEER) is a population-based cancer registry covering 14% of the U.S. population. PARTICIPANTS: Fifty thousand four hundred sixty breast cancer patients aged 65 and older, of whom 1,935 (3.8%) had a diagnosis of AD before or up to 6 months after cancer diagnosis. MEASUREMENTS: Diagnosis of AD was taken from International Classification of Diseases, Ninth Revision, diagnostic codes accompanying Medicare billing claims between 1992 and 1999. The SEER program reported surgery and radiation. Chemotherapy was taken from Medicare billing records. RESULTS: Subjects with AD were diagnosed with breast cancer at later stages, when tumors were larger and the likelihood of lymph node involvement had increased. Patients with AD had a lower likelihood of surgery (odds ratio (OR)=0.60, 95% confidence interval (CI)=0.46-0.81), radiation (OR=0.31, 95% CI=0.23-0.41), and chemotherapy (OR=0.44, 95% CI=0.34-0.58) than those without AD. CONCLUSION: Overall, AD patients receive less treatment for breast cancer than do comparable female Medicare beneficiaries. Chemotherapy and radiation are administered less frequently to women with AD than to other comparable patients. It is unclear whether suboptimal medical care has an effect on their survival. Further research on the effect of screening and treatment decision-making for these patients is warranted. PMID- 16274371 TI - Bereaved family member perceptions of quality of end-of-life care in U.S. regions with high and low usage of intensive care unit care. AB - OBJECTIVES: To compare the quality of end-of-life care of persons dying in regions of differing practice intensity. DESIGN: Mortality follow-back survey. SETTING: Geographic regions in the highest and lowest deciles of intensive care unit (ICU) use. PARTICIPANTS: Bereaved family member or other knowledgeable informants. MEASUREMENTS: Unmet needs, concerns, and rating of quality of end-of life care in five domains (physical comfort and emotional support of the decedent, shared decision-making, treatment of the dying person with respect, providing information and emotional support to family members). RESULTS: Decedents in high- (n=365) and low-intensity (n=413) hospital service areas (HSAs) did not differ in age, sex, education, marital status, leading causes of death, or the degree to which death was expected, but those in the high-intensity ICU HSAs were more likely to be black and to live in nonrural areas. Respondents in high-intensity HSAs were more likely to report that care was of lower quality in each domain, and these differences were statistically significant in three of five domains. Respondents from high-intensity HSAs were more likely to report inadequate emotional support for the decedent (relative risk (RR)=1.2, 95% confidence interval (CI)=1.0-1.4), concerns with shared decision-making (RR=1.8, 95% CI=1.0-2.9), inadequate information about what to expect (RR=1.5, 95% CI=1.3 1.8), and failure to treat the decedent with respect (RR=1.4, 95% CI=1.0-1.9). Overall ratings of the quality of end-of-life care were also significantly lower in high-intensity HSAs. CONCLUSION: Dying in regions with a higher use of ICU care is not associated with improved perceptions of quality of end-of-life care. PMID- 16274372 TI - Optimizing the use of antithrombotic therapy for atrial fibrillation in older people: a pharmacist-led multidisciplinary intervention. AB - OBJECTIVES: To develop, implement, and evaluate a pharmacist-led multidisciplinary intervention in a hospital setting that would optimize antithrombotic use in elderly atrial fibrillation patients. The hypothesis that there would be an increase in the proportion of patients receiving antithrombotic therapy at discharge was tested. DESIGN: Evidence-based algorithms were developed to define the criteria (stroke risk vs contraindications) by which an elderly patient's requirement for antithrombotic therapy was assessed. SETTING: A major Sydney teaching hospital. PARTICIPANTS: Two hundred eighteen consecutively admitted elderly patients (mean age 85.2) were recruited over a 6-month period. INTERVENTION: A pharmacist-coordinated multidisciplinary review process was implemented to coordinate risk assessments and subsequently recommend appropriate antithrombotic therapy, as per the algorithms. MEASUREMENTS: The proportion of patients receiving antithrombotic therapy was assessed on admission (preintervention), at discharge (postintervention), and postdischarge (follow-up at 3 and 6 months). RESULTS: As a result of the intervention, 78 patients (35.8%) required changes to their existing antithrombotic therapy. Of these changes, 60 (76.9%) were "upgrades" to more-effective treatment options (e.g., from no therapy to any agent or from aspirin to warfarin). The remaining 18 (23.1%) changes were "downgrades" to less-effective, albeit safer, options. Despite a significant increase in anti thrombotic use overall (59.6% vs 81.2%, P<.001), fewer patients received warfarin postintervention, after having been assessed as inappropriate candidates (20.7% vs 17.4%, P=.39). CONCLUSION: A pharmacist-led multidisciplinary process was successfully developed and implemented within the hospital setting to increase overall antithrombotic use. Having addressed some of the known barriers and limitations to warfarin use, these algorithms may allow allied health workers, patients, and clinicians to work collaboratively to achieve optimal and, importantly, appropriate (i.e., safe and effective) antithrombotic use in at-risk elderly patients. PMID- 16274373 TI - Effect of acetaminophen on behavior, well-being, and psychotropic medication use in nursing home residents with moderate-to-severe dementia. AB - OBJECTIVES: To evaluate the effect of regularly scheduled administration of analgesic medication on behavior, emotional well-being, and use of as-needed psychotropic medications in nursing home residents with moderate-to-severe dementia. DESIGN: Randomized, double-blind, placebo-controlled, crossover trial. SETTING: Nursing-home based. PARTICIPANTS: Twenty-five nursing home residents with moderate-to-severe dementia. INTERVENTION: Participants received 4 weeks of acetaminophen (3,000 mg/d) and 4 weeks of placebo. MEASUREMENTS: Behavior and emotional well-being were assessed using Dementia Care Mapping, an observational method that quantifies time spent in behaviors across 26 domains (e.g., social interaction, unattended distress) and assesses emotional state while behaviors are being observed. Agitation was measured using the Cohen-Mansfield Agitation Inventory. As-needed psychotropic medication use was aggregated from medication logs. RESULTS: Participants spent more time in social interaction, engaged with media, talking to themselves, engaged in work-like activity, and experiencing unattended distress when they received acetaminophen than they did when they received placebo. Participants also spent less time in their rooms, less time removed from the nursing home unit, and less time performing personal care activities when they received acetaminophen. There were no effects on agitation, emotional well-being, or as-needed psychotropic medication use. CONCLUSION: Untreated pain inhibits activity in nursing home residents with moderate-to severe dementia. Pain treatment in this group may facilitate engagement with the environment. PMID- 16274374 TI - Potential for alcohol and prescription drug interactions in older people. AB - OBJECTIVES: To examine the patterns and prevalence of concomitant alcohol and alcohol-interactive (AI) drug use in older people. DESIGN: Cross-sectional analysis of survey and prescription claims data. SETTING: The Pennsylvania Pharmaceutical Assistance Contract for the Elderly (PA-PACE) program, a state funded program providing prescription benefits to older people with low to moderate incomes. PARTICIPANTS: A total of 83,321 PA-PACE cardholders (age range 65-106) who were using any prescription medications at the time of survey completion. MEASUREMENTS: All AI drugs were identified using a database of medication warning labels obtained from First DataBank. Prescription drug claims were used to characterize AI drug exposure according to therapeutic class of prescription drug use. A mail survey of PA-PACE cardholders was used to examine alcohol use, as well as sociodemographic and health factors associated with concomitant use of alcohol and AI drugs. RESULTS: Seventy-seven percent of all prescription drug users were exposed to AI medications, with significant variation in exposure and concomitant alcohol use according to therapeutic class. Overall, 19% of AI drug users reported concomitant alcohol use, compared with 26% of non-AI drug users (P<.001). Multinomial logistic regression analyses showed that certain groups of older people, including younger older people, men, and those with higher educational levels, were at greater risk for concomitant exposure to alcohol and AI drugs. CONCLUSION: Many older people use alcohol in combination with AI prescription drugs. Clinicians should warn every patient who is prescribed an AI drug about alcohol-drug interactions, especially those at high risk for concomitant exposure. PMID- 16274375 TI - An evaluation of an intervention to assist primary care physicians in screening and educating older patients who use alcohol. AB - OBJECTIVES: To evaluate whether providing physicians and older patients with personalized reports of drinking risks and benefits and patient education reduces alcohol-related risks and problems. DESIGN: Prospective comparison study. SETTING: Community primary care. PARTICIPANTS: Twenty-three physicians and 665 patients aged 65 and older. INTERVENTION: Combined report, in which six physicians and 198 [corrected] patients received reports of patients' drinking classifications and patients also received education; patient report, in which 245 patients received reports and education, but their five physicians did not receive reports; and usual care. MEASUREMENTS: Assessments at baseline and 12 months later to determine patients' nonhazardous (no known risks), hazardous (risks for problems), or harmful (presence of problems) classifications using the Computerized Alcohol-Related Problems Survey (CARPS). The CARPS contains a scanned screening measure and scoring algorithms and automatically produces patient and physician reports and patient education. RESULTS: At baseline, 21% were harmful drinkers, and 26% were hazardous drinkers. The patient report and combined report interventions were each associated with greater odds of lower risk drinking at follow-up than usual care (odds ratio=1.59 and 1.23, respectively, P<.05 for each). The patient report intervention significantly reduced harmful drinking at follow-up from an expected 21% in usual care to 16% and increased nonhazardous drinking from 52% expected in usual care to 58%. Patients in the combined report intervention experienced a significantly greater average decrease in quantity and frequency. CONCLUSION: Older primary care patients can effectively reduce their alcohol consumption and other drinking risks when given personalized information about their drinking and health. PMID- 16274376 TI - Cardiovascular risk factors predictive for survival and morbidity-free survival in the oldest-old Framingham Heart Study participants. AB - OBJECTIVES: To examine whether midlife cardiovascular risk factors predict survival and survival free of major comorbidities to the age of 85. DESIGN: Prospective community-based cohort study. SETTING: Framingham Heart Study, Massachusetts. PARTICIPANTS: Two thousand five hundred thirty-one individuals (1,422 women) who attended at least two examinations between the ages of 40 and 50. MEASUREMENTS: Risk factors were classified at routine examinations performed between the ages of 40 and 50. Stepwise sex-adjusted logistic regression models predicting the outcomes of survival and survival free of morbidity to age 85 were selected from the following risk factors: systolic and diastolic blood pressure, total serum cholesterol, glucose intolerance, cigarette smoking, education, body mass index, physical activity index, pulse pressure, antihypertensive medication, and electrocardiographic left ventricular hypertrophy. RESULTS: More than one third of the study sample survived to age 85, and 22% of the original study sample survived free of morbidity. Lower midlife blood pressure and total cholesterol levels, absence of glucose intolerance, nonsmoking status, higher educational attainment, and female sex predicted overall and morbidity-free survival. The predicted probability of survival to age 85 fell in the presence of accumulating risk factors: 37% for men with no risk factors to 2% with all five risk factors and 65% for women with no risk factors to 14% with all five risk factors. CONCLUSION: Lower levels of key cardiovascular risk factors in middle age predicted overall survival and major morbidity-free survival to age 85. Recognizing and modifying these factors may delay, if not prevent, age-related morbidity and mortality. PMID- 16274377 TI - Is physical activity counseling effective for older people? A cluster randomized, controlled trial in primary care. AB - OBJECTIVES: To establish the effectiveness of the Green Prescription physical activity counseling program in increasing activity and quality of life in older community-dwelling people. DESIGN: Post hoc subgroup analysis of a large cluster randomized, controlled trial. SETTING: One hundred seventeen doctors in 42 primary care practices (74% participation rate) in the Waikato region of New Zealand. PARTICIPANTS: Two hundred seventy sedentary primary healthcare patients aged 65 and older (67% participation rate). INTERVENTION: Patients in intervention practices prompted their primary care doctors or practice nurse to deliver brief activity counseling. A "Green Prescription" was written involving the negotiation of activity goals. Trained exercise specialists from a regional sports foundation gave follow-up telephone support over 3 months. MEASUREMENTS: Leisure moderate and vigorous physical activity, total energy expenditure, systolic and diastolic blood pressure, health-related quality of life, musculoskeletal injuries, falls, and hospitalizations. RESULTS: After 12 months of follow-up, leisure time moderate activity increased by 0.67 h/wk more in the intervention group than the control group (95% confidence interval (CI)=0.17 1.17) and energy expenditure increased by 2.67 kcal/kg per week (95% CI=0.87 4.47) more. For intervention group participants, vitality and general health scales of the 36-item Short Form showed statistically and clinically relevant improvements, and there was a decrease in hospitalizations (P<.03). There were no observable changes in blood pressure, injuries, or falls as a result of the Green Prescription program. CONCLUSION: This physical activity intervention improved activity, energy expenditure, health-related quality of life, and hospitalizations for older primary care patients. Systematic inclusion of the Green Prescription in routine primary health care will probably lead to health gain for older people. PMID- 16274378 TI - A longitudinal study of idiopathic exudative lymphocytic pleural effusion in older people. AB - OBJECTIVES: To assess the long-term prognosis of older patients with idiopathic exudative lymphocytic pleural effusion. DESIGN: Prospective observational study. SETTING: A university-affiliated tertiary care center. PARTICIPANTS: Forty-seven consecutive patients (aged 74.9+/-5.4) with idiopathic exudative lymphocytic pleural effusion were enrolled over a 42-month period. MEASUREMENTS: Baseline sociodemographic information, clinical data, and Charlson Comorbidity Index score were obtained. After an exhaustive examination, clinical evaluation and periodic chest radiographs were taken until one of the endpoints was met: complete resolution of the pleural effusion, death from all causes, or the end of the study period. RESULTS: The mean follow-up period was 16.3+/-17.0 months. During the course of the study, complete resolution of the pleural effusion occurred in 17% of the patients, whereas it remained stable in 45%, and progressed in 38%. In seven cases, the cause of the effusion was established after an average of 84 days, and in another two, the diagnosis was made postmortem. Malignancy was documented in eight of the nine cases. Although the burden of comorbidities and cardiac function at baseline were similar in the three categories, the 3-year survival rate was 63%, 5%, and 0%, respectively. None of the patients developed active tuberculosis, although 15% had positive tuberculin test. CONCLUSION: By categorizing the presence of idiopathic effusion into resolving, persistent, or progressive, this study may provide a more practical approach to the long-term prognosis of older patients with idiopathic exudative lymphocytic effusion who refuse or are considered too frail to undergo an invasive procedure. PMID- 16274379 TI - Age-dependent differences in presentation, risk factor profile, and outcome of suspected acute coronary syndrome. AB - OBJECTIVES: To compare the presenting complaint, risk factors, and outcome of suspected acute coronary syndrome (ACS) in those aged 65 and older with those of a younger cohort. DESIGN: Prospective observational cohort study. SETTING: A typical Scottish district general hospital covering a population of 150,000. PARTICIPANTS: Patients presenting with suspected ACS (N=869) over a 6-month period. MEASUREMENTS: Main presenting complaint and major risk factors including electrocardiogram (ECG) changes. Primary outcome measures were percutaneous coronary intervention, recurrent myocardial infarction, and death at 3-month follow-up. RESULTS: Four hundred seventy-seven (55%) were aged 65 and older. Older patients were less likely to present with chest pain and more likely to present with breathlessness or collapse. They had fewer major risk factors for heart disease. There was a higher proportion with ischemic ECG changes, elevated troponin, and major acute coronary events at follow-up. Older patients were less likely to be accepted for angiography even though they were more likely than the younger cohort to have significant coronary artery disease when angiography was performed (chi-square test, P<.01 for all above). CONCLUSION: Older patients with suspected ACS were more likely to present atypically and have worse outcomes than their younger counterparts, despite having fewer major risk factors. The results highlight the importance of age as a predictor of adverse outcome and suggest that clinicians need to ensure equitable access to angiography for older patients. PMID- 16274380 TI - Mild cognitive impairment and 10-year trajectories of disability in the Iowa Established Populations for Epidemiologic Studies of the Elderly cohort. AB - OBJECTIVES: To apply diagnostic criteria for mild cognitive impairment (MCI) to a geographically representative sample, to estimate the prevalence of MCI, and to estimate 10-year trajectories of incident disability for cognitively intact participants and subgroups with MCI. DESIGN: Prospective cohort; 10 years of follow-up. SETTING: Community-based survey of noninstitutionalized population aged 65 and or older in two rural Iowa counties (Washington and Iowa). PARTICIPANTS: Iowa Established Populations for Epidemiologic Studies of the Elderly (aged > or = 65; N = 3,673; 61.3% female; 99.9% white). MEASUREMENTS: Age, sex, education, Short Portable Mental Status Questionnaire (SPMSQ), 20-item word recall, activities of daily living (ADLs), instrumental activities of daily living (IADLs), chronic medical conditions. RESULTS: MCI was prevalent in 24.7% of participants at baseline. Most participants in the overall cohort remained stable or changed slowly (< or = 1 new limitations) over 10 years (63.1% for SPMSQ, 89.3% for word recall, and 61.7% for ADL disability). For MCI/no prevalent IADL disability (Stage 1 MCI), disability progression was similar to that in the cognitively intact subgroup (median = 0.08 vs 0.05 disabilities per year). For MCI plus prevalent IADL disability (Stage 2 MCI), the median rate of change was equivalent to that of the severely impaired (0.23 disabilities per year; interquartile range = 0.12-0.36). CONCLUSION: Unlike participants with MCI who reported no IADL limitations, those with such limitations were more likely to develop ADL disability--a prerequisite for a diagnosis of dementia. PMID- 16274381 TI - Effects of ultra-low-dose estrogen therapy on muscle and physical function in older women. AB - OBJECTIVES: To determine the effects of ultra-low-dose hormone therapy on muscle mass and physical function in community-dwelling women. DESIGN: Double-blind, placebo-controlled trial. SETTING: Clinical research center in Connecticut. PARTICIPANTS: Healthy, community-dwelling women aged 65 and older (n=167). INTERVENTION: Eligible women were randomly assigned to treatment with 0.25 mg 17 beta estradiol or placebo for 36 months. All women (estradiol or placebo) with an intact uterus received micronized progesterone 100 mg/d for 2 weeks every 6 months. All participants received 1,300 mg elemental calcium with 1,000 IU vitamin D per day. MEASUREMENTS: Appendicular skeletal muscle mass (ASM), lean body mass (LBM), and percentage body fat were measured using dual x-ray absorptiometry. Sarcopenia was defined as skeletal muscle mass (ASM/height2) 2 standard deviations or less than young, healthy reference population mean. Physical activity (Physical Activity Scale in the Elderly (PASE)) and performance were measured. Serum estrone, estradiol, and sex hormone-binding globulin were measured. RESULTS: The prevalence of sarcopenia at baseline was 13%. There were no baseline differences between groups except for PASE score and chair rise time, in which the estrogen group had better performance. No changes in ASM, LBM, percentage of body fat, or physical performance were found after 3 years of estrogen therapy. CONCLUSION: Sarcopenia was present in 13% of this group of community-dwelling, postmenopausal older women. Ultra-low-dose estrogen therapy neither improves nor harms ASM. Similarly, no changes in body fat or physical performance were detected. PMID- 16274382 TI - Validity of the SF-36 five-item Mental Health Index for major depression in functionally impaired, community-dwelling elderly patients. AB - OBJECTIVES: To examine criterion and construct validity of the five-item Mental Health Index (MHI-5) of the 36-item Short Form health survey (SF-36) in relation to the presence of major depression in functionally impaired, community-dwelling elderly patients and of eight subsamples defined by cognitive functioning, levels of functional impairment, and proxy report versus self-report. DESIGN: Cross sectional observational. SETTING: Nineteen counties in western New York, West Virginia, and Ohio. PARTICIPANTS: One thousand four hundred forty-four functionally impaired, community-dwelling Medicare beneficiaries aged 65 and older who participated in the Medicare Primary and Consumer-Directed Care Demonstration. MEASUREMENTS: MHI-5, Mini-International Neuropsychiatric Interview Major Depressive Episode (MINI-MDE) module. RESULTS: The MHI-5 demonstrated sufficient criterion validity (area under the receiver operating characteristic curve=0.837; sensitivity=78.7% and specificity=72.1% using a cutpoint of 59/60) with respect to the presence of depression for the entire sample. A significant correlation between MHI-5 scores and presence of major depression as identified using the MINI-MDE (Spearman correlation=-0.426, P<.001), a strong correlation between the MHI-5 and the SF-36 role emotional scale (Spearman correlation=0.522) and a weak correlation with the SF-36 physical functioning scale (Spearman correlation=0.133) provided evidence for construct validity. Additional evidence is provided by decline in mean MHI-5 score as level of formal education and number of close friends and relatives decreased. All eight subsamples demonstrated similar criterion and construct validity. A Cronbach alpha of 0.794 demonstrated internal consistency reliability. CONCLUSION: This study provides evidence for adequate criterion and construct validity of the MHI-5 in relation to the presence of major depression among functionally impaired, community dwelling elderly Medicare patients. PMID- 16274383 TI - Nursing home practitioner survey of diagnostic criteria for urinary tract infections. AB - OBJECTIVES: To identify clinical and laboratory criteria used by nursing home practitioners for diagnosis and treatment of urinary tract infections (UTIs) in nursing home residents. To determine practitioner knowledge of the most commonly used consensus criteria (i.e., McGeer criteria) for UTIs. DESIGN: Self administered survey. SETTING: Three New Haven-area nursing homes. PARTICIPANTS: Physicians (n=25), physician assistants (PAs, n=3), directors/assistant directors of nursing (n=8), charge nurses (n=37), and infection control practitioners (n=3). MEASUREMENTS: Open- and closed-ended questions. RESULTS: Nineteen physicians, three PAs, and 41 nurses completed 63 of 76 (83%) surveys. The five most commonly reported triggers for suspecting UTI in noncatheterized residents were change in mental status (57/63, 90%), fever (48/63, 76%), change in voiding pattern (44/63, 70%), dysuria (41/63, 65%), and change in character of urine (37/63, 59%). Asked to identify their first diagnostic step in the evaluation of UTIs, 48% (30/63) said urinary dipstick analysis, and 40% (25/63) said urinalysis and urine culture. Fourteen of 22 (64%) physicians and PAs versus 40 of 40 (100%) nurses were aware of the McGeer criteria for noncatheterized patients (P<.001); 12 of 22 (55%) physicians and PAs versus 38 of 39 (97%) nurses used them in clinical practice (P<.001). CONCLUSION: Although surveillance and treatment consensus criteria have been developed, there are no universally accepted diagnostic criteria. This survey demonstrated a distinction between surveillance criteria and criteria practitioners used in clinical practice. Prospective data are needed to develop evidence-based clinical and laboratory criteria of UTIs in nursing home residents that can be used to identify prospectively tested treatment and prevention strategies. PMID- 16274384 TI - B-type natriuretic peptide is associated with mortality in older functionally impaired patients. AB - OBJECTIVES: To determine the predictive power of B-type natriuretic peptide (BNP) regarding death in older, functionally impaired patients with multiple comorbidity. DESIGN: Prospective cohort study. SETTING: Specialist geriatric assessment clinic and day hospital. PARTICIPANTS: Two hundred ninety-nine older, functionally impaired patients, mean age 79 at enrollment. MEASUREMENTS: Full clinical history and examination, baseline BNP, and echocardiography. Date and cause of death were ascertained from Scottish death records. Kaplan-Meier survival curves were constructed for quartiles of log (BNP), and the contribution of BNP to prediction of death was investigated. RESULTS: The follow-up period ranged from 3.9 to 5.2 years (mean 4.4 years). BNP was a powerful independent predictor of all-cause and cardiovascular mortality. BNP was a more powerful predictor than blood pressure, diabetes mellitus, smoking, echocardiographic left ventricular hypertrophy, left ventricular systolic dysfunction, or age. BNP predicted death in those with and without a previous cardiovascular event at baseline. CONCLUSION: BNP has significant predictive power for death in older, functionally impaired patients. PMID- 16274385 TI - Weight loss, muscle strength, and angiotensin-converting enzyme inhibitors in older adults with congestive heart failure or hypertension. AB - OBJECTIVES: To determine whether angiotensin-converting enzyme (ACE) inhibitor use may be associated with weight maintenance and sustained muscle strength (measured by grip strength) in older adults. DESIGN: Data from the Cardiovascular Health Study (CHS), a community-based prospective cohort study of 5,888 older adults, were used. SETTING: Subjects were recruited from four U.S. sites beginning in 1989; this analysis included data through 2001. PARTICIPANTS: CHS participants with congestive heart failure (CHF) or treated hypertension. MEASUREMENTS: The exposure, current ACE inhibitor use, was ascertained by medication inventory at annual clinic visits; the outcomes were weight change and grip-strength change during the following year. Multivariate linear regression was used, accounting for correlations between observations on the same participant over time. RESULTS: The average annual weight change was -0.38 kg in 2,834 participants (14,443 person-years) with treated hypertension and -0.62 kg in 342 participants (980 person-years) with CHF. ACE inhibitor use was associated with less annual weight loss after adjustment for potential confounders: a difference of 0.17 kg (95% confidence interval (CI)=0.05-0.29) in those with treated hypertension and 0.29 kg (95% CI=-0.25-0.83) in those with CHF. There was no evidence of association between ACE inhibitor use and grip-strength change. CONCLUSION: ACE inhibitor use may be associated with weight maintenance, but not maintenance of muscle strength, in older adults with treated hypertension. PMID- 16274386 TI - Mortality risk and prospective medicare expenditures for persons with dementia. AB - OBJECTIVES: To determine whether an index that stratifies dementia patients into high- and low-mortality-risk groups can be effective in prospectively identifying persons with high and low expenditures. DESIGN: A mortality risk index developed for persons with dementia was used to stratify persons into groups at high and low risk of 12-month mortality. Expenditures were compiled from Medicare claims for 12 months after the predicted mortality classification. SETTING: The Medicare Alzheimer's Disease Demonstration Evaluation (MADDE) operated in eight sites across the United States. Subjects were living in a community setting at time of their mortality risk classification. PARTICIPANTS: Participants (N=2,255) were volunteer enrollees in MADDE; all had a diagnosis of irreversible dementia and two or more limitations in activities of daily living (ADLs). MEASUREMENTS: Outcomes were Medicare Part A expenditures. Risk factors included age, sex, ADL limitations, selected chronic conditions, prior year hospital stays, and relationship to caregiver. RESULTS: Patients predicted as at high risk of death had higher average monthly Medicare expenditures than those predicted to be at low risk of death. At the 50th sensitivity percentile for mortality, differences in average expenditures exceeded 700 dollars per month. CONCLUSION: It is not known whether it is possible to achieve reductions in expenditures for patients with advanced dementia at high risk of mortality, but study findings support consideration of an advanced illness model that would attempt to generate acute services savings and use these savings for palliative services that might improve patients' and caregivers' quality of life. PMID- 16274387 TI - Effect of a mandatory geriatric medicine clerkship on third-year students. AB - A nationwide push has increased geriatric medicine instruction within medical school curricula. Some institutions have proceeded with an integrated 4-year curriculum while others have constructed discrete courses in the third or fourth year of medical school. This paper describes the impact of a new mandatory 4-week geriatric medicine clerkship on third-year students developed by the Donald W. Reynolds Department of Geriatric Medicine at the University of Oklahoma Health Sciences Center. In the first year of implementation, 135 students took the course on both the Oklahoma City and Tulsa campuses. Clinical sites included inpatient, VA extended care unit, outpatient clinics, dementia clinics, home care, long-term care settings, and hospice. Didactic instruction used formal lectures and problem-based learning. The impact of the clerkship on students was assessed in three areas: knowledge, skills, and attitude using a pre- and postknowledge test, student satisfaction survey, and written comments. This article discusses how the clerkship resulted in increased knowledge of geriatric medicine. Student self-report indicates that the clerkship enhanced clinical evaluation and patient assessment skills. Students indicated that the experience was positive and recognized the importance of geriatric medicine in their development as doctors. PMID- 16274388 TI - Impediments to timely diagnosis of Alzheimer's disease in African Americans. AB - The purpose of this study was to identify early patterns of care for Alzheimer's disease (AD) in a cohort of African-American patients and their caregivers presenting at an inner city clinic and a suburban memory assessment clinic. Caregivers (N=79) of patients diagnosed with probable AD were interviewed. Data were collected about the delay from noticing first AD signs until recognition that a problem existed and delay from problem recognition until first physician consultation. Patients and caregivers had lower educational status, and patients had been diagnosed more recently at the inner city clinic than at the suburban clinic, although MMSE scores of patients at the two clinics did not differ; median delays in caregivers' recognizing a problem and in consulting a physician were also similar across clinics. Delay was as long as 7 years between noticing symptoms and problem recognition and between problem recognition and physician consultation. Although patients attending the suburban clinic were more likely to have previously seen a physician than those attending the inner city clinic, they were no more likely to have received a prior diagnosis of AD. Lack of physician contact is likely to be widespread in families caring for African Americans with AD. Physician consultation is more characteristic of more highly educated families but may not yield a correct diagnosis for the patient. Intensive efforts are needed to connect African-American families with physicians and to achieve more timely diagnosis of AD to enable families to understand the illness, plan for patient safety, and make long-term plans. PMID- 16274389 TI - Geriatrics in Brazil: a big country with big opportunities. AB - Brazil has approximately 180 million inhabitants, of whom 15.2 million are aged 60 and older and 1.9 million are aged 80 and older. By 2025, the Brazilian elderly population is expected to grow to more than 32 million. Brazil has many problems related to its geographic and population size. Great distances between major cities, marked cultural and racial heterogeneity between the various geographic regions, high poverty levels, and decreasing family size all combine to put pressure on the medical and social services that can be made available to the elder population. Less than 500 Brazilian physicians are certified as geriatricians, translating into one geriatrician for every 37,000 elderly Brazilians. Beside 15 geriatric medicine residencies a larger number of fellowship programs exist, and these programs are in high demand, with more than 20 candidates per position, indicating new opportunities for growth in elder care. In addition, geriatric initiatives such as the annual elder vaccination program and the elder statute, recently approved by the Brazilian Congress, indicate that geriatric care in Brazil is entering a new era of growth and development. Although the challenges remain great, there are opportunities for Brazilian geriatrics and gerontology. PMID- 16274390 TI - No old man ever forgot where he buried his treasure: concepts of cognitive impairment in old age circa 1700. AB - Cognitive impairment in old age is one of the most important topics in modern geriatrics. This article discusses the historical dimensions of this phenomenon. To this end, a number of primary sources ranging from antiquity to the modern era are evaluated. Although a physiology and pathology of old age were conceptualized in Greco-Roman times, cognitive impairment in old age remained a marginal issue until the 17th century. Alternatively, after 1500, medicine boasted detailed theories on the physiology and pathology of old age. There are several possible explanations for this unusual situation. Underlying conflict between idealistic and materialistic views of man played a decisive role, for these concepts differed considerably regarding the intellectual and mental functioning of the soul as well as the effects of the passage of time. After Cartesianism and Iatromechanism had pushed these traditional boundaries back, the problem of cognitive impairment in old age was increasingly regarded as a physical illness and began to receive more attention. Just as its philosophical and theological context shaped early modern medicine, contemporary nonmedical disciplines such as genetics, (neuro-)biology, and the information sciences influence modern research. PMID- 16274391 TI - Urinary incontinence: an important and underrecognized complication of type 2 diabetes mellitus. PMID- 16274392 TI - Pleiotropic effects of angiotensin-converting enzyme inhibitors and the future of cachexia therapy. PMID- 16274395 TI - Compliance after colorectal cancer screening and health-related quality of life domains in geriatric Japanese. PMID- 16274397 TI - A case report of paranoid delusion with galantamine use. PMID- 16274396 TI - Vitamin B as a predictor of mortality in elderly patients. PMID- 16274398 TI - Improvement of heart rate recovery after exercise training in older people. PMID- 16274399 TI - Etidronate ameliorates painful soft-tissue calcification in Werner syndrome. PMID- 16274400 TI - Hypoadiponectinemia in bedridden female patients younger than 75. PMID- 16274401 TI - Acute cholecystitis as a presentation of metastatic breast carcinoma of the gallbladder: a case report. PMID- 16274402 TI - Diagnostic accuracy of telehealth community dementia assessments. PMID- 16274403 TI - A comparison of gait assessment methods: Tinetti and GAITRite electronic walkway. PMID- 16274404 TI - How patients experience psoriasis: results from a European survey. AB - BACKGROUND: Most of the psoriasis that affects the European population is treated with topical preparations. Compliance rates, however, are known to be quite low. OBJECTIVE: To provide an update on how psoriasis is experienced by patients and how well these patients comply with prescribed topical treatment. METHODS: This was an observational and transversal survey conducted in 1281 patients with psoriasis. Patient input alone formed the basis of the study. Participants were contacted through the national psoriasis patient associations in France, the United Kingdom, Belgium, Germany and the Netherlands. RESULTS: The study showed that 32% of the patients suffer currently from psoriasis in the face, skin folds and genital areas--areas very sensitive to topical treatments. Most of the patients (74%) considered their psoriasis as at least moderately severe, and 73% stated not to comply with their current treatment. Lack of efficacy and messiness of the treatment were the main reasons for non-compliance, an element that is especially important for the use in sensitive areas such as the face, skin folds and genitalia. CONCLUSION: The present survey confirmed that cosmetic acceptability is, together with efficacy and safety, a very important aspect of a successful treatment of psoriasis. PMID- 16274406 TI - Topical use of calcitriol 3 microg/g ointment in the treatment of mild-to moderate psoriasis: results from an open-label study. AB - BACKGROUND: Calcitriol is the active metabolite and hormonal analogue of vitamin D3. It is widely used for the topical treatment of psoriasis showing good tolerability and effectiveness. OBJECTIVES: To assess the efficacy, tolerability and safety of calcitriol 3 microg/g ointment in mild to moderate plaque psoriasis involving sensitive areas. METHODS: Sixty patients with a body surface area < 35% were enrolled into a prospective open label clinical study. Patients were treated for 12 weeks with a twice daily calcitriol 3 microg/g topical ointment application. Efficacy and safety were assessed during the therapy and during a 6 month follow-up period. RESULTS: The study demonstrated a high clinical remission rate which progressively increased throughout therapy (11.6% at week 4, 28.3% at week 8 and 63.3% at week 12). No serious adverse events and clinically relevant changes of calcium/phosphorus homeostasis were reported. CONCLUSIONS: The study suggests that calcitriol 3 microg/g ointment, applied twice daily, is an effective topical treatment for chronic psoriatic plaques involving less than 35% of the body surface and sensitive areas. PMID- 16274407 TI - The right dose in the right place: an overview of current prescription, instruction and application modalities for topical psoriasis treatments. AB - BACKGROUND: Compared to treatment success with oral medications, treatment success with topical treatments is currently difficult to achieve. AIM: To assess and compare existing methods of lesion size evaluation, prescription habits, instruction and application modalities of topical medications. METHODS: Review of current and new procedures to estimate the body surface area, to calculate the most adequate quantity of medication to be prescribed, to instruct patients and to assess the individual dosing procedures of topical treatments. RESULTS: The most accurate method to assess the body surface area remains the rule of nine, allowing, together with a recently developed calculation disc, to estimate the most adequate quantity of topical treatment (here calcitriol 3 g/g ointment) to be prescribed. Precise instructions to the patient are other important elements for the successful treatment of dermatoses. Dosing devices, such as a spatula, may help patients to dose their daily topical treatment, avoiding over- and underdosing of the medication. CONCLUSION: A correct evaluation of the lesions sizes, a precise calculation of the treatments quantity to be prescribed, clear instructions and treatment-specific individual dosing devices may help to achieve higher patient compliance and treatment success. PMID- 16274405 TI - Rheological properties of three different vitamin D ointments and their clinical perception by patients with mild to moderate psoriasis. AB - BACKGROUND: Ointments, classically used for the treatment of dermatological diseases, are monophasic viscous semisolid formulations. According to the proportion of their compounds, they have physicochemical and organoleptic properties and when applied on skin show a specific behaviour allowing to be spread more or less easily. OBJECTIVE: To measure in vitro rheological characteristics of three vitamin D derivative ointments prescribed for the treatment of psoriasis, and to compare their viscosity and clinical acceptability when applied on the diseased skin. METHODS: Rheological characteristics of tacalcitol 4 microg/g, calcipotriol 50 microg/g and calcitriol 3 microg/g ointments were assessed by measuring the oscillatory viscoelastic parameters and the permanent flow analysis. Clinical acceptability was studied in 20 psoriatic male or female subjects, aged 18 years or older. A survey evaluated the acceptability of calcitriol vs. tacalcitol and calcipotriol. Questions included information about fluidity, spreading capacity and stickiness after application. RESULTS: We demonstrated that viscoelastic parameters were four times higher for ointment tacalcitol than for calcipotriol and calcitriol, corresponding to a higher consistency of ointment tacalcitol compared to calcipotriol and calcitriol showing both similar results; better fluidity was demonstrated by calcitriol than by tacalcitol and calcipotriol. Comparable results were obtained for the quality to be spread. The sensation of stickiness, significantly different between tacalcitol and calcitriol, was not different between calcipotriol and calcitriol. CONCLUSION: The above results confirm the relationship between rheological in vitro and sensorial in vivo results: variations between different formulations may have an important influence on non-adherence and treatment failure. PMID- 16274411 TI - Our search for the porcine mother rotor. AB - A single stationary mother rotor, located in the fastest activating region and giving rise to activation fronts that propagate throughout the remainder of the myocardium, has been hypothesized to be responsible for the maintenance of ventricular fibrillation (VF). Others have reported a mother rotor in guinea pigs and rabbits. We wanted to see if a mother rotor exists in a larger heart, that is, pigs. Epicardial mapping studies have demonstrated that VF wavefronts in pigs tend to propagate from the posterior basal LV to the anterior LV and on to the anterior RV, raising the possibility of a mother rotor in the posterior LV. However, no sustained reentry consistent with a mother rotor was found on the posterior LV epicardium, even though an intramural mapping study showed that the fastest activating transmural layer was near the epicardium. Many wavefronts in the posterior LV entered the mapped region from the posterior boundary of the mapping array, adjacent to the posterior descending coronary artery, raising the possibility that a mother rotor is located in the right ventricle or septum. Since a previous study has shown that the RV activates more slowly than the LV during VF, the more likely site for a mother rotor was the septum. However, we then performed a study in which we recorded from the right side of the septum and found that reentry was uncommon there also and that the activation rate was slower than the posterobasal LV. Many of the VF wavefronts in the septum passed from the posterior septum toward the anterior septum. This fact coupled with the fact that many wavefronts passed from the posterior LV free wall toward the anterior LV free wall point to the region where the posterior free wall intersects with the septum, the region where the posterior papillary muscle is located, as the possible site of a mother rotor. Indeed, a recent abstract by others reports that, after propranolol, a stable reentrant circuit is present on the endocardium at the insertion of the posterior papillary muscle into the LV free wall in pigs. PMID- 16274412 TI - Cardiac resynchronization--a heart failure perspective. AB - Over the past 15-20 years the development of new heart failure pharmacologic therapy has lowered mortality by 30-40% for this serious and prevalent clinical syndrome, within clinical trials conducted in patients with a dilated cardiomyopathy phenotype. However, over the past 5 years progress in the development of additional effective drugs has slowed, in part due to the success of neurohormonal inhibitors, on which background new therapies must be developed. That there is not an absolute ceiling on the development of new heart failure therapies has been convincingly recently demonstrated in electrophysiologic device trials, conducted on the background of maximal neurohormonal inhibition. Two trials, COMPANION and CARE-HF, have demonstrated unambiguously that in advanced heart failure patients with a marker of mechanical intraventricular dyssynchrony, increased QRS duration, cardiac resynchronization therapy in the form of biventricular pacing can improve major clinical outcomes including mortality. In addition, COMPANION also demonstrated that the addition of an ICD further improved mortality reduction, by lowering the incidence of sudden death. These trials indicate that device/drug therapy is at least additive in the treatment of heart failure, and they herald a new era in the multi-modality approach to therapeutics. PMID- 16274413 TI - Relationship of the implantable cardioverter defibrillator and chronic resynchronization therapy: the perfect marriage? AB - BACKGROUND: The two major modes of death in the patient with a reduced ejection fraction (EF) are death due to heart failure and death due to lethal arrhythmia, essentially the two sides of the same coin. Over the last 20 years, two therapies cardiac resynchronization therapy (CRT) and the implantable cardioverter defibrillator (ICD)-have been developed and tested in clinical trials. They are now, in conjunction with appropriate medical therapy, the mainstays of therapy for these two commonly encountered clinical problems. METHOD AND RESULTS: Both of these therapies were conceived and patented by two Baltimore cardiologists, Michel Mirowski and Morton Mower (Table I). The path to everyday acceptance of both therapies was remarkably similar. The concept and early success of both devices was accomplished but the proof of concept depended on a series of carefully designed randomized clinical trials that showed that both the CRT and ICD devices saved lives in the low EF population, especially when used together. These trials overcame substantial skepticism on behalf of elements of the cardiology and electrophysiology establishment. CONCLUSION: We are now at a crossroads in the further extension of either therapy. The majority of the indications for either device alone or in combination are established. In the next few years, assuming the continued commitment on the part of regulatory agencies to fully embrace evidence-based medicine, we will see indications extended but only by the careful clinical trials that became the bedrock of their initial acceptance. PMID- 16274414 TI - Multicenter automatic defibrillator implantation trial-cardiac resynchronization therapy (MADIT-CRT): design and clinical protocol. AB - The planned MADIT-CRT trial is designed to determine if CRT-D will reduce the risk of mortality and HF events by approximately 25% in subjects with ischemic (NYHA class I-II) and non-ischemic (NYHA class II) cardiomyopathy, left ventricular dysfunction (EFor=130 ms). PMID- 16274415 TI - Cardiac resynchronization therapy: the MGH experience. AB - Cardiac resynchronization therapy (CRT) has gained acceptance as a useful form of device therapy for patients with refractory congestive heart failure. Despite recent technical advances, a significant number of patients continue to remain unresponsive to this form of therapy. This article provides an overview of CRT, highlights several unresolved issues and describes ongoing research efforts to address some of these important questions. PMID- 16274416 TI - CRT-D therapy in patients with left ventricular dysfunction and atrial fibrillation. AB - The number of patients with atrial fibrillation and congestive heart failure has steadily increased in the United States. The presence of atrial fibrillation increases morbidity and mortality for patients with left ventricular dysfunction. The emergence of cardiac resynchronization therapy to improve symptoms and survival from congestive heart failure may provide benefits for those with atrial fibrillation; we review the pathophysiology of atrial fibrillation in the presence of left ventricular dysfunction and the promise of cardiac resynchronization therapy to improve symptoms for the for these patients. PMID- 16274417 TI - Safety and efficacy of flecainide in subjects with Long QT-3 syndrome (DeltaKPQ mutation): a randomized, double-blind, placebo-controlled clinical trial. AB - BACKGROUND: We conducted a study of chronic therapy with flecainide versus placebo in a small group of LQT-3 patients with the DeltaKPQ deletion to evaluate the safety and efficacy of flecainide in this genetic disorder. In vitro studies have shown that flecainide provides correction of the impaired inactivation associated with the DeltaKPQ deletion. METHODS: A randomized, double-blind, placebo-controlled clinical trial was conducted with flecainide and placebo in six male LQT-3 subjects with the DeltaKPQ deletion. RESULTS: The lowest possible dose of flecainide associated with at least a 40 ms reduction in the QTc interval was determined in an initial open-label, dose-ranging investigation using one fourth or half of the recommended maximal antiarrhythmic flecainide dose. QTc reduction was achieved with a flecainide dose of 1.5 mg/kg per day in 4 subjects and with 3.0 mg/kg per day in 2 subjects. Subjects were randomized to four 6 month alternating periods of flecainide and placebo therapy based on the open label dose findings. Average QTc values during placebo and flecainide therapies were 534 ms and 503 ms, respectively, with an adjusted reduction in QTc of -27.1 ms (95% confidence interval: -36.8 ms to -17.4 ms; P<0.001) at a mean flecainide blood level of 0.11+/-0.05 microg/ml. Minimal prolongation in QRS occurred (mean: +2.5 ms), and there were no major adverse cardiac effects. CONCLUSIONS: Chronic low-dose flecainide significantly shortens the QTc interval in LQT-3 subjects with the DeltaKPQ mutation. No major adverse drug effects were observed with flecainide during this trial, but the sample size is not large enough to evaluate the safety of flecainide therapy in patients with this mutation. PMID- 16274418 TI - Cost-effectiveness of implanted defibrillators in young people with inherited cardiac arrhythmias. AB - BACKGROUND: The implanted cardioverter-defibrillator (ICD) has been shown to improve survival in adult patients with high risk acquired cardiac disease, with a cost-effectiveness ratio in the range of $30,000 to $185,000 per quality adjusted-life-year saved. However, data on the benefit and cost-effectiveness of device therapy in high-risk patients with inherited cardiac disorders are limited. METHODS: We developed two separate computer-based analytical models to compare non-ICD with ICD therapy in patients (age range: 10-75 years) with long QT syndrome (LQTS) and hypertrophic cardiomyopathy (HCM). In each disease entity patients were stratified into low-risk (no known risk factors); high-risk (known risk factors [primary prevention]); and very high-risk (prior near-fatal events [secondary prevention]). Net costs were defined as the difference between costs resulting from treatment of the disease and savings due to gained productivity attributable to prevention of sudden cardiac death. Outcome was defined as costs per quality-adjusted life-years saved. RESULTS: In LQTS, defibrillator therapy was shown to be cost effective in high-risk male patients (incremental cost effectiveness ratio [ICER]=$3328 per quality-adjusted-life-year saved), and cost saving in high-risk females (ICER=$7102 gained per quality-adjusted-life-year saved) and very high-risk males and females (ICER=$15,483 and 19,393 gained per quality-adjusted-life-year saved, respectively). In HCM, defibrillator therapy was cost saving in both male and female high-risk (ICER=$17,892 and $17,526 gained per quality-adjusted-life-year saved, respectively) and very high-risk (ICER $22,944 and $22,329 gained per quality-adjusted-life-year saved, respectively) patients. Defibrillator therapy was not shown to be cost effective in low-risk patients with either LQTS or HCM (ICER in the range of $400,000 to $600,000 lost per quality-adjusted-life-year saved). Sensitivity analyses were consistent with the results in each risk group. CONCLUSIONS: In appropriately selected patients with inherited cardiac disorders, early intervention with ICD therapy is cost-effective to cost saving due to added years of gained productivity when the lifespan of an individual at risk is considered. PMID- 16274419 TI - Quantitative analysis of production traits in saltwater crocodiles (Crocodylus porosus): I. reproduction traits. AB - Repeatability and phenotypic correlations were estimated for saltwater crocodile reproductive traits. No pedigree information was available to estimate heritability or genetic correlations, because the majority of breeder animals on farms were wild-caught. Moreover, as the age of the female breeders could not be accounted for, egg-size measurements were used as proxies. The reproductive traits investigated were clutch size (total number of eggs laid), number of viable eggs, number of eggs that produced a live, healthy hatchling, hatchability, average snout-vent length of the hatchlings and time of nesting. A second data set was also created comprising binary data of whether or not the female nested. Repeatability estimates ranged from 0.24 to 0.68 for the measurable traits, with phenotypic correlations ranging from -0.15 to 0.86. Repeatability for whether a female nested or not was 0.58 on the underlying scale. Correlations could not be estimated between the measurement and binary traits because of confounding. These estimates are the first published for crocodilian reproduction traits. PMID- 16274420 TI - Quantitative analysis of production traits in saltwater crocodiles (Crocodylus porosus): II. age at slaughter. AB - Crocodile morphometric (head, snout-vent and total length) measurements were recorded at three stages during the production chain: hatching, inventory [average age (+/-SE) is 265.1 +/- 0.4 days] and slaughter (average age is 1037.8 +/- 0.4 days). Crocodile skins are used for the manufacture of exclusive leather products, with the most common-sized skin sold having 35-45 cm in belly width. One of the breeding objectives for inclusion into a multitrait genetic improvement programme for saltwater crocodiles is the time taken for a juvenile to reach this size or age at slaughter. A multivariate restricted maximum likelihood analysis provided (co)variance components for estimating the first published genetic parameter estimates for these traits. Heritability (+/-SE) estimates for the traits hatchling snout-vent length, inventory head length and age at slaughter were 0.60 (0.15), 0.59 (0.12) and 0.40 (0.10) respectively. There were strong negative genetic (-0.81 +/- 0.08) and phenotypic (-0.82 +/- 0.02) correlations between age at slaughter and inventory head length. PMID- 16274421 TI - Pedigree information reveals moderate to high levels of inbreeding and a weak population structure in the endangered Catalonian donkey breed. AB - The Catalonian donkey is one of the most endangered donkey breeds in the world. At present, five main subpopulations exist: AFRAC, which consists of many genetically connected Catalonian localities; Berga, which consists of a single herd located also in Catalunya but under private management; and three minor non Catalonian subpopulations (Huesca, Sevilla and Toledo). In this study, we analysed the pedigree information of the Catalonian donkey herdbook to assess the genetic diversity and population structure of the breed. We found that the Catalonian donkey has suffered an important loss of genetic diversity and moderate to high increases of inbreeding because of the abuse of a few individuals in matings. This scenario is mainly characterized by the fact that both the effective number of founders and ancestors for the whole population was 70.6 and 27, respectively, while the equivalent number of founders was 146.5 and the number of ancestors explaining overall genetic variability was 93. In addition, only 14% of animals born between the 1960s and 1970s were significantly represented in the pedigree. Our results also show that subpopulations where breeders exchanged reproductive individuals had low levels of inbreeding and average relatedness. One subpopulation, Berga, was reproductively isolated and showed high levels of inbreeding (F = 7.22%), with average relatedness (AR = 6.61%) playing an important role in increasing the values of these coefficients in the whole pedigree. Using genealogical F-statistics we have found little evidence of population structuring (F(ST) = 0.0083) with major genetic differences among non-Catalonian subpopulations. PMID- 16274422 TI - A genotype probability index for multiple alleles and haplotypes. AB - We use linear algebra to calculate an index of information content in genotype probabilities which has previously been calculated using trigonometry. The new method can be generalized allowing the index to be calculated for loci with more than two alleles. Applications of this index include its use in genotyping strategies, strategies to manage genetic disorders and in estimation of genotype effects. PMID- 16274423 TI - Genetic and environmental parameters for racing time at different distances in Brazilian Thoroughbreds. AB - The aim of the present study was to investigate genetic parameters for racing time in Thoroughbred horses racing at distances between 1000 and 1600 m subdivided into 100-m intervals. The data provided by TURFETOTAL Ltda comprised races that occurred in the Gavea and Cidade Jardim race tracks over a period of 11 years (1992-2002) and consisted of 32,145 races and 238,890 time records. The variance components necessary to obtain the heritability and repeatability estimates of the traits studied were estimated with the MTDFREML program, and animal age at race (3 years old or younger, 4, 5 and older than 5 years), sex (male and female), number of races (1-32,145), and postposition at start (1-11) as fixed effects, and animal and permanent environmental random effects were included in a one-trait animal model. Males were significantly superior to females at all distances. Excluding the 1100 m distance, animals 4 years of age were significantly faster than the mean of the other ages for all distances analysed. Horses older than 5 years showed a significantly lower performance than the mean of the other ages for all distances analysed, except for the 1100 m. Postpositions one and two did not differ significantly from one another for any of the distances analysed. These two inner positions both together varied from the other positions depending on race length. The components of additive genetic and permanent environmental variance varied in a similar way, tending to decrease with increasing racing distance, and the other temporary environmental variance almost doubled from 1000 to 1600 m. As was the case for the additive genetic and environmental variances, heritability and repeatability estimates tended to decrease with increasing distance, indicating that selection based on racing time will be less successful when the racing distance increases. PMID- 16274424 TI - Effect of the polymorphism of prolactin receptor (PRLR) and leptin (LEP) genes on litter size in Polish pigs. AB - The aim of the experiment was to use the DNA mutations in the PRLR and LEP genes to determine associations between the genotype and litter size in Polish Large White x Landrace sows. Reproductive traits investigated were: total number of piglets born (TNB), number of piglets born alive (NBA) and number of piglets weaned. The polymorphism in PRLR and LEP genes was detected using the polymerase chain reaction-restriction fragment-length polymorphism (PCR-RFLP) method, with specific primers and the restriction enzymes AluI and HinfI respectively. Two different alleles of PRLR and LEP gene were identified: alleles A (0.62) and B (0.38) of the PRLR gene and alleles C (0.10) and T (0.90) of the LEP gene. The relationships between the PRLR and LEP genotypes and TBN, NBA and NW were analysed. The analysis showed, in first parity sows, statistically significant (p < or = 0.01) differences between sows carrying different PRLR genotypes. In later parities, sows with the AA genotype still had the largest litter size compared with AB and BB sows, but the difference was statistically not significant. Analysis of the interaction PARITY x PRLR showed small and statistically not significant differences. The analysis of relationship between different LEP genotypes and TNB, NBA, NW showed small and statistically non-significant differences. PMID- 16274425 TI - Estimates of genetic diversity in the brown cattle population of Switzerland obtained from pedigree information. AB - The study investigates the genetic diversity present as well as its development in the Brown Cattle population of Switzerland from pedigree information. The population consisted of three subpopulations, the Braunvieh (BV), the original Braunvieh (OB) and the US-Brown Swiss (BS). The BV is a cross of OB with BS where crossing still continues. The OB is without any genetic influence of BS. The diversity measures effective population size, effective number of ancestors (explaining 99% of reference genome) and founder genome equivalents were calculated for 11 reference populations of animals born in a single year from 1992 onwards. The BS-subpopulation consisted of animals and their known ancestors which were used in the crossing scheme and was, therefore, quite small. The youngest animals were born in 2002, the oldest ones in the 1920s. Average inbreeding was by far the highest in BS, in spite of the lowest quality of pedigrees, and lowest in OB. Effective population size obtained from the difference between average inbreeding of offspring and their parents was, mostly due to the heavy use of few highly inbred BS-sires, strongly overestimated in some BV-reference populations. If this parameter was calculated from the yearly rate of inbreeding and a generation interval of 5 years, no bias was observed and ranking of populations from high to low was OB-BV-BS, i.e. equal to the other diversity parameters. The high genetic diversity found in OB was a consequence of the use of many natural service sires. Rate of decrease of effective number of ancestors was steeper in BV than OB was, however, equal for founder genome equivalents. Founder genome equivalents were more stable than effective population sizes calculated from the difference between average inbreeding of offspring and parents. The five most important ancestors contributed one-third of the 2002-reference genomes of BV and OB, in BV all were BS-sires. The relative amount of BS-genes in the BV-genome increased from 59.2% to 78.5% during the 11 years considered. PMID- 16274426 TI - Polymorphism of 5'-region of the bovine growth hormone receptor gene. AB - Genes coding for growth hormone (GH) and GH receptor (GHR) are candidates for quantitative trait markers in farm animals. This work describes a search for nucleotide sequence polymorphisms within the 5'-region of the bovine GHR gene. Two new single nucleotide polymorphisms were found: restriction fragment length polymorphisms (RFLPs) at a Fnu4HI/TseI site (C/T transition at position -1104), and at a Sau96I site (C/T transition at position -262). The Fnu4HI/TseI polymorphic site is located within the 1.2-kbp LINE-1 retrotransposon upstream of the P1 promoter, while the Sau96I RFLP locates in the P1 promoter for exon 1A. The appearance of the Sau96I RFLP was studied in representatives of two bovine species, Bos taurus and Bos indicus. An absolute correlation was observed between Sau96I genotype and the insertion/deletion of LINE-1. PMID- 16274427 TI - Single nucleotide polymorphism detection in promoter III of the acetyl-CoA carboxylase-alpha gene in sheep. AB - Three novel SNPs were identified in the locus OAR292286, encoding the DNA sequence of promoter III of the ovine acetyl-CoA carboxylase-alpha gene, in Italian sheep of four breeds: Gentile di Puglia (25 individuals) and Sopravissana (31) which are triple-purpose local endangered breeds, Comisana (25) which is a local non-selected, non-endangered dairy breed and Sarda (15) which is a popular selected high yielding dairy breed. Variant alleles are: G/T at 1330 bp, C/G at 1338 bp and C/T at 1430 bp. Frequencies of the variant alleles were calculated and chi-squared analysis of the differences in allele frequency between breed pairs was performed. Allele frequencies of the Sarda breed differ significantly from the other considered breeds. PMID- 16274430 TI - Mother-child conversations about pictures and objects: referring to categories and individuals. AB - The distinction between individuals (e.g., Rin-Tin-Tin) and categories (e.g., dogs) is fundamental in human thought. Two studies examined factors that influence when 2- to 3-year-old children and adults focus on individuals versus categories. Mother-child dyads were presented with pictures and toys (e.g., a picture of a boat or a toy boat). Conversations were coded for references to generic categories ("Dogs are furry"), ostensive labels ("This is a dog"), or specific individuals ("Lassie"). Overall, pictures generated more talk about categories; objects generated more talk about individuals. However, when objects could not be manipulated, speakers expressed relatively more category references. These results suggest that representations (in the form of pictures or objects-on display) encourage young children and parents alike to think about categories. PMID- 16274431 TI - The role of parenting styles in children's problem behavior. AB - This study investigated the combination of mothers' and fathers' parenting styles (affection, behavioral control, and psychological control) that would be most influential in predicting their children's internal and external problem behaviors. A total of 196 children (aged 5-6 years) were followed up six times from kindergarten to the second grade to measure their problem behaviors. Mothers and fathers filled in a questionnaire measuring their parenting styles once every year. The results showed that a high level of psychological control exercised by mothers combined with high affection predicted increases in the levels of both internal and external problem behaviors among children. Behavioral control exercised by mothers decreased children's external problem behavior but only when combined with a low level of psychological control. PMID- 16274432 TI - Prospective relations among victimization, rejection, friendlessness, and children's self- and peer-perceptions. AB - This study investigated the prospective links between three forms of peer adversities (i.e., victimization, rejection, and lack of reciprocated friendships) and children's perceptions of themselves and of their peers. The sample consisted of 212 children (107 boys and 105 girls, 11-13 years) recruited from four primary schools and followed up for a period of one year. The results showed that a negative self-perception was a risk factor for the development of all forms of peer adversities. Of the three forms of peer adversities assessed, victimization and rejection had an influence on children's peer perceptions. None of the peer adversities predicted changes in self-perceptions. The results partially support a transactional model between children and their environments. PMID- 16274433 TI - Pathways from prematurity and infant abilities to later cognition. AB - This study examined the relation of information processing in 7-month-old preterms (<1750 g at birth) and full-terms to Bayley Mental Development Indexes (MDIs) at 2 and 3 years. The infant measures were drawn from four cognitive domains: attention, speed, memory, and representational competence. Structural equation modeling showed that these measures of infant information processing mediated the effects of prematurity, and that there was a cascade of effects, with infant processing speed influencing memory and representational competence, which in turn influenced later MDI. This study shows that infant information processing mediates the effect of prematurity on later cognition, and delineates pathways whereby infant abilities relate to one another and to later outcome. PMID- 16274434 TI - The nominal passover effect depends on addressee age, speaker goal, and object similarity. AB - If after teaching a label for 1 object, a speaker does not name a nearby object, 3-year-olds tend to reject the label for the nearby object (W.E. Merriman, J.M. Marazita, L.H. Jarvis, J.A. Evey-Burkey, and M. Biggins, 1995a). In Studies 1 (5 year-olds) and 3 (3-year-olds), this effect depended on object similarity. In Study 2, when a speaker used a label without teaching it, 5-year-olds showed no passover effect. 3-year-olds showed none for inanimate objects, but one for animate objects. When extraneous factors that may have promoted animate object individuation were eliminated (Study 3), 3-year-olds showed the effect when a label was taught, but not when it was merely used. Children honor rational restrictions on when the unacceptability of a name can be inferred from its nonoccurrence. PMID- 16274435 TI - Development of temporal-reconstructive abilities. AB - In a study of the ability to reconstruct the times of past events, 86 children from 4 to 13 years recalled the times of 2 in-class demonstrations that had occurred 3 months earlier and judged the times of hypothetical events. Many of the abilities needed to reconstruct the times of events were present by 6 years, including the capacity to interpret many temporally relevant cues, but there were substantial changes well into middle childhood in the availability of temporally useful episodic information. Children were poor at remembering the events' proximity or order with respect to a major holiday, but the order of the 2 target events was well recalled by 6 years. PMID- 16274436 TI - Analysis and interpretation of twin studies including measures of the shared environment. AB - Recent reports using a classical behavior genetic research design in which twin data are combined with a measured characteristic of their shared family environment have made striking claims about estimating environmental influences on behavior with genetic effects controlled. Such claims are overstated for two related reasons. First, when a variable is measured at the family level in a way that makes it necessarily equivalent for twins reared together, it is not possible to partition it into genetic and environmental components. Second, although structural equation modeling and DeFries-Fulker analysis are sound tools for the analysis of many types of twin data, they do not control for genetic or environmental confounds when estimating the effect of measured family-level variables. PMID- 16274437 TI - Physical discipline and children's adjustment: cultural normativeness as a moderator. AB - Interviews were conducted with 336 mother-child dyads (children's ages ranged from 6 to 17 years; mothers' ages ranged from 20 to 59 years) in China, India, Italy, Kenya, the Philippines, and Thailand to examine whether normativeness of physical discipline moderates the link between mothers' use of physical discipline and children's adjustment. Multilevel regression analyses revealed that physical discipline was less strongly associated with adverse child outcomes in conditions of greater perceived normativeness, but physical discipline was also associated with more adverse outcomes regardless of its perceived normativeness. Countries with the lowest use of physical discipline showed the strongest association between mothers' use and children's behavior problems, but in all countries higher use of physical discipline was associated with more aggression and anxiety. PMID- 16274438 TI - A monozygotic twin differences study of nonshared environmental influence on adolescent depressive symptoms. AB - The monozygotic (MZ) twin differences method was used to examine nonshared environmental aspects of the association of parenting (punitive and constructive discipline), negative life events (independent and dependent), and peers (antisocial and prosocial) with adolescent depressive symptoms in terms of sequelae, risk, and maintaining factors. Two self-report booklets were completed by 328 MZ twin pairs aged 12-19 years, 6 months apart. Increased punitive discipline and negative life events were found to be relevant nonshared environmental factors that followed depression. Negative life events were the only factor that had a direct nonshared environmental association with concurrent depressive symptoms in adolescents independent of previous depressive symptoms, and all other study variables. PMID- 16274439 TI - Preschoolers mistrust ignorant and inaccurate speakers. AB - Being able to evaluate the accuracy of an informant is essential to communication. Three experiments explored preschoolers' (N=119) understanding that, in cases of conflict, information from reliable informants is preferable to information from unreliable informants. In Experiment 1, children were presented with previously accurate and inaccurate informants who presented conflicting names for novel objects. 4-year-olds-but not 3-year-olds-predicted whether an informant would be accurate in the future, sought, and endorsed information from the accurate over the inaccurate informant. In Experiment 2, both age groups displayed trust in knowledgeable over ignorant speakers. In Experiment 3, children extended selective trust when learning both verbal and nonverbal information. These experiments demonstrate that preschoolers have a key strategy for assessing the reliability of information. PMID- 16274440 TI - Differences in the relations between antisocial behavior and peer acceptance across contexts and across adolescence. AB - This study tests the hypothesis that, during adolescence, antisocial behavior becomes positively associated with peer acceptance. This hypothesis was tested considering both classroom and out-of-class peer relations. Data from a previously published study, with a cross-sectional sample of 577 Italian 11- to 13-year-olds, were used. Analyses showed that in the 6th grade antisocial behavior was negatively related to classroom peer preference, but not significantly related to out-of-class peer inclusion. By the 8th grade, antisocial behavior was positively related to out-of-class peer inclusion, but not significantly related to classroom peer preference. Similar results were found for males and females. The higher level of peer acceptance among the 8th grade antisocial individuals was primarily due to nominations received by other antisocial individuals. PMID- 16274441 TI - Out of the toolbox: toddlers differentiate wobbly and wooden handrails. AB - This study examined whether 16-month-old walking infants take the material composition of a handrail into account when assessing its effectiveness as a tool to augment balance. Infants were encouraged to cross from one platform to another via bridges of various widths (10, 20, 40 cm) with either a "wobbly" (foam or latex) or a wooden handrail available for assistance. Infants attempted to walk over wider bridges more often than narrow ones, and attempts were more frequent when the sturdy wooden handrail was available. Infants tailored their exploratory behaviors, bridge-crossing strategies, and handrail-use strategies to the material properties of the rail. PMID- 16274442 TI - Personality and daily experience revisited. AB - In this introduction to the Journal of Personality special issue devoted to "Advances in Personality and Daily Experience" we attempt to trace recent progress in the study of personality and daily life. We begin by comparing and contrasting the methods of daily experience research adopted by investigators who authored empirical articles in the 1991 and current special issues according to the mode, schedule, and sampling frame for data collection. We then compare data analytic strategies across the 14 years between the two special issues and conclude that although the nature of the questions addressed has not changed dramatically, newer analytic methods and associated software have enhanced our ability to answer these questions more precisely. Finally, we provide an overview of the special issue contributions organized around three broad themes: personal vulnerabilities and resources that predict reactions to everyday events, interpersonal manifestations of personality in daily experience, and how personality in daily life affects physical and mental health and relates to disease risk. PMID- 16274443 TI - The daily life of the garden-variety neurotic: reactivity, stressor exposure, mood spillover, and maladaptive coping. AB - This article describes a series of studies using the daily process paradigm to describe and understand the affective dynamics of people who experience frequent and intense bouts of a wide range of negative emotions. In several studies, community residents reported on problem occurrence and affect several times a day or at the end of the day. We found reliable evidence that persons who scored high (vs. low) in Neuroticism reported more daily problems, tended to react with more severe emotions, experienced more mood spillover from prior occasions, and exhibited stronger reactions to recurring problems (the "neurotic cascade"). The susceptibility of neurotics to stress seems to extend to all types of problems while certain other dimensions of personality (e.g., Agreeableness) are associated with hyperreactivity to particular kinds of problems. The research demonstrates how daily process research can provide insight about classic problems in the field of individual differences. PMID- 16274445 TI - Distinguishing affective and non-affective reactions to daily events. AB - The study of daily events has been dominated by a focus on affective reactions to daily events. Although informative, this research needs to be complemented by research on non-affective and cognitive reactions to events. Although daily events are certainly related to how people feel, they are also related to how people think, particularly about themselves. The present article presents the results of a series of studies examining relationships between daily events and both affective and non-affective states. These results suggest that although affective and non-affective reactions to daily events may covary (e.g., when people feel badly, they may think more poorly about themselves and vice versa), this covariation is not perfect. Non-affective states covary with daily events above and beyond the covariation between events and affect, and affective states covary with events above and beyond the covariation between events and non affective states. PMID- 16274444 TI - Dynamic approaches to emotions and stress in everyday life: Bolger and Zuckerman reloaded with positive as well as negative affects. AB - A decade ago, Bolger and Zuckerman (1995) incorporated personality into the study of daily life events and psychological distress. Their approach put an entirely new cast on research and theorizing in this area. In their work, they focused on the predominantly negative personality trait of Neuroticism. In this article we extend their work to include theory and measurement of positive events and indicators of well-being. Integrating these research strands offers the possibility of a comprehensive yet highly sensitive and dynamic approach to the study of emotions, stress, and health in everyday life. PMID- 16274446 TI - Measuring self-esteem in context: the importance of stability of self-esteem in psychological functioning. AB - In this article, I report on a research program that has focused on the joint roles of stability and level of self-esteem in various aspects of psychological functioning. Stability of self-esteem refers to the magnitude of short-term fluctuations that people experience in their current, contextually based feelings of self-worth. In contrast, level of self-esteem refers to representations of people's general, or typical, feelings of self-worth. A considerable amount of research reveals that self-esteem stability has predictive value beyond the predictive value of self-esteem level. Moreover, considering self-esteem stability provides one way to distinguish fragile from secure forms of high self esteem. Results from a number of studies are presented and theoretical implications are discussed. PMID- 16274447 TI - Unfolding interpersonal behavior. AB - Models of interpersonal traits have traditionally contained two independent dimensions, one referring to dominance as the opposite of submissiveness and the other referring to agreeableness as the opposite of quarrelsomeness. These models are primarily based on psychometric analyses of the co-occurrence of interpersonal characteristics. The present article reviews literature based on event-contingent recording studies that examine whether the structure of interpersonal behavior as revealed in its everyday occurrence is consistent with this model of interpersonal traits. Evidence from studies of the effects of hierarchical social role situations, the relations between behaviors and affect, and the effects of alterations in serotonin are used to evaluate whether dominance, submissiveness, agreeableness, and quarrelsomeness are related, opposite, or independent behavioral systems. The pattern of findings suggests that agreeableness and quarrelsomeness may be part of the same behavioral system while dominance and submissiveness may have separate underlying behavioral systems. PMID- 16274448 TI - Coping in context: the role of stress, social support, and personality in coping. AB - Personality and social relationships play an important role in almost every aspect of stress and coping. Daily process methods are particularly useful in elucidating how these factors might influence both responses to and outcomes of stress. Our work has linked both dimensions of personality, particularly the Big Five, and aspects of social relationships, particularly social support, to the likelihood of engaging in certain coping strategies and the effectiveness or outcomes of these coping strategies. In addition, we have found the effect of personality on coping and stress outcomes to vary by the situational context in which stress occurs. We review findings from our recent daily process studies of stress, coping, and social support. Further, we discuss the costs and benefits of the daily process methodology for addressing these questions, highlighting the clinical utility of findings gleaned with the use of this approach. Finally, we discuss future directions and applications of daily process methods to the study of stress and coping. PMID- 16274449 TI - A daily process approach to individual differences in stress-related alcohol use. AB - In our research we have used a variety of daily designs (paper-and-pencil or Internet-based daily diaries, experience sampling via palmtop computers) to examine how individual differences in coping styles, alcohol-outcome expectancies, and drinking motives predict stress-related and negative affect related alcohol use. Our work has furthered research in this area in two ways. First, we have examined how these risk factors are associated with within-person associations among temporally proximal reports of stressors, negative affect, and alcohol use. Second, we have examined whether purportedly stable individual difference factors demonstrate meaningful within-person variation and thus also might be conceptualized as important process variables. Our findings from this line of research, along with those from new data that we present, indicate some disparities between the results found at the within-person versus the between person levels of analysis. More importantly, this line of research offers insights into questions that could not be addressed using traditional cross sectional or long-term longitudinal designs. Limitations and future directions are discussed. PMID- 16274450 TI - Daily affective reactivity as a prospective predictor of depressive symptoms. AB - Daily affective reactivity refers to the within-subject relationship between daily stress and daily mood. Most stress researchers have conceptualized daily affective reactivity as a dependent variable to be predicted by individual difference variables such as personality and psychopathology. In contrast, in our recent research, we have conceptualized daily affective reactivity as an independent variable that can predict depressive symptoms. In this article, we summarize three studies that relied on a daily process methodology and multilevel modeling to assess affective reactivity in the context of daily stressful events. Two of the studies (Cohen, Butler, Gunthert, & Beck, 2005; Gunthert, Cohen, Butler, & Beck, 2005) sampled adult outpatients in cognitive therapy and evaluated the predictive role of daily affective reactivity in treatment outcome (depression reduction). A third study (O'Neill, Cohen, Tolpin, & Gunthert, 2004) evaluated the predictive role of college students' daily affective reactivity in the development of depressive symptoms. We consider the strengths and weaknesses of a daily process methodology for research on depression in both clinical and nonclinical samples. PMID- 16274451 TI - Dynamic influences on smoking relapse process. AB - This article describes a program of research applying Ecological Momentary Assessment (EMA) methods to study relapse to cigarette smoking, with a particular focus on the role of negative affect (NA) and self-efficacy (SE). Day-to-day changes in mood and stress did not predict lapse risk, but more proximal changes in affect were associated with lapses: Many lapses were marked by intense NA and by NA increases in the preceding hours. Individual differences in baseline SE predicted lapse risk, but daily SE was relatively stable during abstinence and did not influence lapse risk. However, lapses resulted in immediate drops in SE, and day-to-day changes in postlapse SE predicted progression to relapse, even after accounting for concurrent smoking. SE showed momentary drops associated with NA, but only among smokers with low baseline SE. Individual differences in baseline SE were only expressed situationally under conditions of NA. The findings highlight the importance of dynamic changes in background conditions and in immediate states as important influences on lapses and relapse and also suggest the importance of considering person by situation interactions. PMID- 16274452 TI - Psychosocial stress and cardiovascular risk: what is the role of daily experience? AB - We describe an ecological momentary assessment (EMA) protocol designed to measure daily life experiences along several psychosocial dimensions (Social Conflict, Task Demand, Decisional Control, Negative Affect, and Arousal) hypothesized to be relevant for cardiovascular disease risk. In a large community sample, these assessments have been administered in conjunction with ambulatory blood pressure (ABP) monitoring and measures of subclinical cardiovascular disease. Several results have emerged that support the promise of this approach. First of all, each of these dimensions of experience appears to be a trigger for cardiovascular activation, with movement on each scale being associated with significant within person changes in heart rate and blood pressure in the natural environment. Second, there appear to be individual differences in physiological responsiveness to these dimensions of daily experience, with such differences being associated in some cases with laboratory-based assessments of cardiovascular reactivity, as might be expected. Third, EMA ratings are associated in several instances with (between-person) stable individual differences in ABP readings. And finally, we have found that some of the characteristics defined by our EMA ratings are related to measures of subclinical atherosclerosis. Such effects appear to be mediated, in part, by ABP. The advantages of using EMA measures for capturing the effects of psychosocial stressors are highlighted by comparing the predictive validity of these daily life assessments to traditional global self-reports. We conclude by describing future plans for use of this type of assessment protocol for helping us to characterize psychosocial characteristics relevant to cardiovascular disease risk. Personality and social processes during daily living may have an important impact not only on psychological functioning but also on physical health and disease. In this article we review our recent work involving the assessment of daily experience as a means of better understanding the role of psychosocial stress in cardiovascular disease risk. First, we outline the rationale for our interest in daily experience sampling for examining these processes, and we describe the methods we have adopted for this purpose. Next, we review findings from a recent study in which we have used these approaches to examine the role of psychosocial stress on ABP and atherosclerosis in a community dwelling sample. We compare our approach to more traditional (global self-report and laboratory-observation) methods for examining these relationships in humans, and we discuss potential future directions for use of daily experience sampling in studies of health and behavior. PMID- 16274453 TI - Mechanisms of blister induction by autoantibodies. AB - Autoimmune diseases are characterized by defined self-antigens, organ specificity, autoreactive T cells and/or autoantibodies that can transfer disease. Autoimmune blistering diseases are organ-specific autoimmune diseases associated with an immune response directed to structural proteins mediating cell cell and cell-matrix adhesion in the skin. While both autoreactive T and B cells have been detected and characterized in patients with autoimmune blistering diseases, current evidence generally supports a pathogenic role of autoantibodies for blister formation. The immunopathology associated with blisters induced by autoantibodies relies on several mechanisms of action. Autoantibodies from patients with pemphigus diseases can exert a direct effect just by binding to their target mediated by steric hindrance and/or by triggering the transduction of a signal to the cell. In most subepidermal autoimmune blistering conditions, in addition to the binding to their target antigen, autoantibodies need to interact with factors of the innate immune system, including the complement system and inflammatory cells, in order to induce blisters. Generally, decisive progress has been made in the characterization of the mechanisms of blister formation in autoimmune skin diseases. However, various aspects, including the exact contribution of steric hindrance and signal transduction for pemphigus IgG induced acantholysis or the fine tuning of the inflammatory cascade triggered by autoantibodies in some subepidermal blistering diseases, still need to be addressed. Understanding the mechanisms by which autoantibodies induce blisters should facilitate the development of more specific therapeutic strategies of autoimmune blistering diseases. PMID- 16274454 TI - Comparison of histologic, biochemical, and mechanical properties of murine skin treated with the 1064-nm and 1320-nm Nd:YAG lasers. AB - The goal of this study was to compare the effects of the Q-switched 1064-nm Nd:YAG laser and the 1320-nm Nd:YAG laser non-ablative treatments on mouse skin in vivo. Skin elasticity measurements were carried out with a Reviscometer, and skin samples were taken for histological study, hydroxyproline content assay and estimation of collagen type I and III. By the second month after non-ablative treatments, the 1064-nm laser treatment resulted in an average of 25% greater improvement of skin elasticity, 6% more increase of dermal thickness, and 11% higher synthesis of hydroxyproline than the 1320-nm laser. Collagen type III increased markedly after the 1064-nm laser treatment whereas more collagen type I was elicited by the 1320-nm laser. Our results demonstrated that the 1064-nm laser was more effective than the 1320-nm Nd:YAG laser in non-ablative treatments, but the results needed to be confirmed in humans. It appeared that photo-mechanic reaction could cause more collagen type III synthesis whereas the photo-thermal effect was in favor of the formation of collagen type I. PMID- 16274455 TI - Comparative genomic hybridization analysis of cutaneous large B-cell lymphomas. AB - The aim of the present study was to identify genetic aberrations in a series of patients with cutaneous large B-cell lymphoma (LBCL) using comparative genomic hybridization (CGH). Eighteen consecutive patients with primary (13 patients) (PCLBCL) and secondary (five patients) (SCLBCL) cutaneous large B-cell lymphoma were included in the study. Nine cases corresponded to PCLBCL leg type and four cases primary cutaneous follicle centre-cell lymphoma (PCFCL). Chromosomal imbalances (CIs) were detected in 14 of 18 samples (77.8%). All of nine cases with PCLBCL leg type and two of four cases with PCFCL showed CIs (100% and 50%, respectively). Regarding SCLBCL, in three of five cases (60%), CIs were detected. The most frequently detected gains involved 2q, 5q, 3 and 7q and amplifications affected 18, 12 and 13. Frequent losses were found in 17p. In PCLBCL leg type, the most frequent gains involved 2q and 7q, amplifications were localized in chromosomes 12, 13 and 18 and losses affected chromosomes 17p and 19. In PCFCL, gains located in 3q, 4 and 7q were found. Our study seems to confirm clear-cut differences between primary cutaneous LBCL and nodal diffuse LBCL, and it suggests the presence of genotypic differences between cases of PCLBCL leg type and cases of PCFCL. PMID- 16274456 TI - A novel splice-site mutation in ECM-1 gene in a consanguineous family with lipoid proteinosis. AB - Lipoid proteinosis (LP) (OMIM 247100) is a rare, autosomal recessive disorder. Recent studies have shown that LP is the result of reduced expression of the extracellular matrix protein gene (ECM-1), in which loss-of-function mutations have been described. In the present report, we describe a large consanguineous family with LP. We identified a homozygous splice-site mutation in intron 1 (IVS1 + 1G-->C) in three clinically affected patients. This is the first splice-site mutation reported in LP and is the most 5' of all ECM-1 mutations described thus far. It is predicted to result in the removal of the translation initiation site, thus ablating all three known ECM-1 isoforms (ECM-1a, ECM-1b, and ECM-1c). In addition, we found a novel splicing variant that is not associated with the disease (DQ010946) and results in the generation of a short, prematurely terminating transcript. This case further emphasizes the role of ECM-1 in LP and highlights the unresolved genotype-phenotype correlation in this disease. PMID- 16274457 TI - Increased scratching counts depend on a decrease in ability of cutaneous prostaglandin D2 biosynthesis in NC/Nga mice with atopic dermatitis. AB - Spontaneous and 2,4,6-trinitrochlorobenzene (TNCB)-induced dermatitis models using NC/Nga mice have been recognized as animal models of atopic dermatitis. We reported that scratching behavior leads to dermatitis in a spontaneous dermatitis but not in a TNCB-induced dermatitis. Prostaglandin D2 (PGD2) suppressed the scratching behavior of NC/Nga mice, suggesting that PGD2 plays a physiological role on inhibiting pruritus. We studied whether there was a difference in skin PG contents between spontaneous and TNCB-induced dermatitis. Spontaneous dermatitis was induced by cohabitation with NC/Nga mice having severe skin lesions. TNCB induced dermatitis was caused by applications of TNCB. PGD2, PGE2, 6keto PGF1alpha, and PGF2alpha contents in the skin were examined using enzyme immunoassay kits. For studying ability to produce skin PGs, PG contents were evaluated after topical treatment of arachidonic acid (AA) or mechanical scratching. In spontaneous dermatitis, PGE2, 6keto-PGF1alpha, and PGF2alpha contents increased with dermatitis, but only PGD2 did not do so. In TNCB-induced dermatitis, PGD2, PGE2, 6keto-PGF1alpha, and PGF2alpha increased. Determination of skin PG contents after AA treatment or mechanical scratching revealed that skin PGD2 production of conventional group of spontaneous dermatitis was lower than the specific pathogen-free group. It seemed that ability of skin PGD2 production was attenuated in spontaneous dermatitis. These results suggest that enhancement of scratching behavior in spontaneous dermatitis was caused by the defect of ability to produce PGD2, which plays a physiological role in inhibiting pruritus, resulting in development of dermatitis. PMID- 16274458 TI - Increased expression of chemokines in the skin of chronic proliferative dermatitis mutant mice. AB - Chemokines direct the migration of leukocytes to sites of inflammation and are potential targets for anti-inflammatory therapy. Chronic proliferative dermatitis (cpdm/cpdm) mutant mice develop a persistent eosinophilic dermatitis associated with increased T(H)2 cytokines in the skin. Expression patterns of chemokines in the skin of cpdm/cpdm mice were evaluated to define the mechanisms driving cutaneous infiltration by leukocytes. RNA isolated from the skin of mutant and littermate control mice revealed a significant increase in Ccl1 (TCA-3), Ccl2 (MCP-1), Ccl11 (eotaxin), Ccl17 (TARC), Cxcl10 (IP-10), and the chemokine receptor Ccr3. The concentration of CCL11 protein was increased two- to threefold in the skin of cpdm/cpdm mice by enzyme-linked immunosorbent assay. In vitro culture of primary dermal fibroblasts from cpdm/cpdm and control mice with tumor necrosis factor, IL-4, and IL-13 stimulation did not reveal differences in their ability to secrete CCL11, suggesting that the increased chemokine expression observed in the skin of cpdm/cpdm mice is most likely caused by the increased T(H)2 cytokines in the dermis of this mouse model. Treatment of cpdm/cpdm mice with CCL11-neutralizing polyclonal antibodies did not affect the number of eosinophils in the skin or the severity of the dermatitis. Neutralizing multiple chemokines or chemokine receptors may be necessary to decrease eosinophil accumulation. The cpdm/cpdm mutant mouse is a potentially useful model to determine the role of various chemokines in eosinophil accumulation in chronic inflammation. PMID- 16274459 TI - The EP3 receptor stimulates ceramide and diacylglycerol release and inhibits growth of primary keratinocytes. AB - Primary human keratinocytes (PHKs) are known to express the EP3 subtype of prostaglandin E2 receptor. To better understand the role of EP3 receptors in regulating epidermal function, we characterized their expression, localization, and signaling effects in human skin. Three different splice variants of the EP3 receptor (EP3A1, EP3C, and EP3D) were found to be expressed. Immunohistochemical analysis of human skin demonstrated that EP3 receptors were most prominently expressed in the basal and lower spinous layers of the epidermis. The EP3 receptor agonist sulprostone was then used to examine EP3 receptor-dependent keratinocyte signaling pathways and functional effects. We observed that sulprostone inhibits keratinocyte growth at doses between 0.02 and 2 nM and induces sn-1,2-diacylglycerol (DAG) and ceramide production. Concurrent expression of the cell-cycle inhibitory protein p21WAF1 also occurred. These data suggest that EP3 receptors produce epidermal growth inhibition through the action of DAG and ceramide second messengers. PMID- 16274460 TI - The status quo and quo vadis of mast cells. AB - Mast cells (MCs) have been intensely investigated over the past two decades, e.g. the numbers of PubMed-listed reports on MCs have steadily increased and doubled over the past twenty years. Surprisingly, many recent findings that have fundamentally changed our understanding of MC biology and functions have yet to be sufficiently recognized by scientists interested in cutaneous biology and clinical dermatologists. The aim of this study is to review recent hallmark contributions to the field of MC research, to outline the development of our current knowledge of MCs, and to predict the outcome of future MC research efforts. The development of straightforward rodent in vivo models has allowed for the identification and characterization of various novel MC functions. MC effects are not limited to the induction of pathology, but can serve important functions in maintaining health and preventing disease. Attempts to better define the role of MCs in the human system may lead to novel strategies for treating inflammatory disorders and could eventually allow us to utilize MCs for improving responses to environmental danger signals. PMID- 16274461 TI - General practictioners' management of cancer in England: secondary analysis of data from the National Survey of NHS Patients-Cancer. AB - Relatively little is understood concerning the exact role of general practice in the cancer patients' pre-diagnostic, and post-diagnostic journey. This paper aims to explore this role using data from the National Survey of NHS Patients-Cancer. Data from 65,192 patients relating to five questions from this survey were analysed in detail with particular relevance to differences between the six cancers [breast, colorectal, lung, ovarian, prostate and non-Hodgkin's lymphoma (NHL)], and socio-demographic variables (age, gender and social class). There were considerable differences between patients with the six cancers, and the role of general practice in the cancer diagnosis, and post-diagnosis management. The vast majority of patients saw their general practitioner (GP) with symptoms prior to being seen in hospital. A significant minority were told their diagnosis by their GP. About half the sample were told to contact their GP post-discharge, and about half did so. Being told to contact the GP post-discharge was strongly associated with actually seeing the GP. Most patients felt that their GP was given enough information about their treatment or condition. In conclusion, this work has quantified the central role of general practice in cancer diagnosis and management in England. There remain considerable resource, educational and research needs to continue to provide high-quality cancer care in primary care. PMID- 16274462 TI - Knowledge of treatment intent among patients with advanced cancer: a longitudinal study. AB - Increasingly, patients with cancer wish to be more fully informed about their disease, treatment and prognosis, and to participate in decision making. The objective of this study was to assess knowledge of diagnosis and goals of treatment among patients with advanced cancer, and also to assess whether this knowledge changed over time. A cohort of 181 subjects with advanced cancer receiving palliative therapies were interviewed at entry and again 12 weeks later. Knowledge of disease diagnosis, treatment intent, and the main sources of information were determined. Twenty per cent of subjects considered their illness to be non-life threatening, and 46% correctly perceived treatment intent as non curative; 29% believed the intent of treatment was cure. Subjects resident in rural areas were more likely to misunderstand the goal of their treatment. Treatment modality was significantly associated with knowledge of treatment intent, and subjects in the last 6 months of life had clearer understanding that treatment intent was non-curative. Many patients with advanced cancer do not understand the goals of treatment. Excessive optimism may lead to impaired decision making. Further empirical research into information transfer and predictors of accurate patient understanding would assist clinicians in their discussions of prognosis and potential treatment outcomes with patients. PMID- 16274463 TI - Family members' experiences, information needs and information seeking in relation to living with a patient with oesophageal cancer. AB - A qualitative study was conducted with the aim to describe family members' experiences, information needs and information seeking in relation to living with a patient suffering from oesophageal cancer. Data were collected by means of semi structured interviews with nine family members. A content analysis was used in order to organize data. It was found that family members were not aware of the severe diagnosis and the illness caused intrusions on the family. The time following diagnosis family members' information seeking was low. They used interpersonal as well as mass media sources to obtain knowledge about and handle the uncertainty related to the illness. Some family members did not actively seek information. In conclusion, healthcare professionals are expected to improve quality of cancer care. These improvements are unlikely to occur without an understanding of family members' needs. This study shows that the family members were unprepared of receiving a diagnosis of oesophageal cancer. They emphasized the importance of including the children in the care given. Moreover, the whole family was faced with uncertainty, which led to intrusion on everyday life. Therefore, family members used different strategies for managing the uncertainty, whereof one was searching for information. The primary source of information was the physician. PMID- 16274464 TI - Clinical usefulness of oral granisetron hydrochloride for alleviation of delayed nausea and vomiting induced by CPT-11. AB - This open label pilot study evaluated the safety and efficacy of the oral 5-HT3 receptor antagonist granisetron for prophylaxis of delayed chemotherapy-induced nausea and vomiting (CINV) in 30 patients with advanced or recurrent colorectal cancer. Patients were studied during two cycles of a 5-week regimen with irinotecan (CPT-11) and UFT. Patients received prophylactic anti-emetic therapy that included intravenous granisetron. If Grade 1 or higher severity gastrointestinal symptoms occurred during 6 days after CPT-11 administration in Cycle 1, then oral granisetron was administered daily for the following 5 days of CPT-11 in Cycle 2. Sixteen patients (53.3%) experienced delayed CINV in Cycle 1. The incidence of Grade 2 or higher vomiting was 32.1% and 27.7% in Cycles 1 and 2 in males (P = 0.554) respectively, and 54.6% and 32.4% in females (P = 0.001) respectively. Granisetron is effective against delayed Grade 2 or higher vomiting induced by CPT-11/UFT in female patients, although granisetron alone may not sufficiently control nausea induced by this regimen. PMID- 16274465 TI - Pulmonary embolism after transcatheter arterial chemoembolization. AB - Metastatic hepatic tumours can be treated with hepatic transcatheter arterial chemoembolization (TACE). Common complications associated with TACE include hepatic insufficiency, fever, and pain. However, pulmonary embolism is rarely documented as a fatal adverse effect. We report a case of pulmonary embolism following TACE in a renal cell carcinoma patient with liver metastases. Total recovery is noted after the effective treatment. PMID- 16274466 TI - Naturopathy/herbalism consultations by mid-aged Australian women who have cancer. AB - Complementary and alternative medicine (CAM) is now a significant practice issue for those delivering cancer care with a range of CAM being utilized by a significant number of patients with cancer. While various studies have explored the prevalence of CAM use among cancer patients, little is currently known about naturopathy/herbalism use by patients with cancer in Australia. This paper reports the prevalence of naturopath/herbalist consultations among mid-aged Australian women with cancer. The research was conducted as part of the Australian Longitudinal Study on Women's Health, with the data for this analysis coming from the third survey of 11,202 women aged 50-55, conducted in 2001. For all cancers combined, 15.7% of women with cancer were found to consult a naturopath/herbalist. Mid-aged women with cancer were found to be more likely to consult a naturopath/herbalist than mid-aged women without cancer. Naturopathy/herbalism consultations appear to be utilized by the women with cancer alongside and as a supplement to conventional health services. Given the prevalence of consultations with herbal therapists/naturopaths among mid-aged women with cancer in Australia, it is important that physicians and others involved in cancer patient care and management are informed and educated about naturopathy/herbalism and its use among their patients. PMID- 16274467 TI - An evaluation of the introduction of Clinical Trial Officer roles into the cancer clinical trial setting in the UK. AB - The National Cancer Research Network (NCRN) was created in 2001 to improve the infrastructure for cancer research within the National Health Service (NHS) in the UK and ensure that research is better integrated with cancer care. The NCRN consists of 34 regional networks which map onto the 34 cancer networks that were established following the publication of the NHS Cancer Plan with its aim of improving the coordination and delivery of cancer care and treatment nationally. An objective of the NCRN is to increase recruitment into cancer trials through improved support. One cancer research network responded by introducing Clinical Trial Officers (CTOs) into the cancer clinical trial setting in order to combine the range of tasks required to support clinical trials into a single role with the ultimate aim of increasing recruitment into cancer clinical trials. Evaluation during the first 14 months of their introduction assessed the impact of the new CTO role on cancer trial recruitment, its acceptability to those involved in trials and its effectiveness in achieving increased recruitment. Evaluation identified appropriate induction and training programmes required to support these new roles and the identification of a model for the introduction of CTO posts in other networks across the country. The findings presented in this paper identify that CTO roles can effectively be introduced into a cancer network and have an impact on recruitment to clinical trials within that network. The data collected provided an in-depth insight into how these roles were perceived, have developed and what supporting structures need to be in place to enable them to flourish. Recruitment in the network has increased and there has been a raising of awareness of clinical trials and cancer research across the whole of the cancer network where the CTOs were based. We conclude that CTO roles can offer a creative alternative to staffing cancer clinical trial units and that such a model could be introduced across other networks. Similar models could also be introduced into other disease settings where clinical research is taking place. PMID- 16274468 TI - Effects of Qi therapy (external Qigong) on symptoms of advanced cancer: a single case study. AB - The aim of this study was to examine the effectiveness of Qi therapy (external Qigong) in the management of symptoms of advanced cancer in a man. We used a single case study design to evaluate the effectiveness of Qi therapy (external Qigong) in a 35-year-old man with advanced cancer (Stage IV) involving metastases in the stomach, lung and bone (Karnofsky performance scale: KPS, 40: requires special care and assistance, disabled). Treatment involved six days of pre assessment, eight treatment sessions on alternate days over 16 days, and a two week follow-up phase. A visual analogue scale (VAS) was used to assess the patient's self-reported symptoms of cancer over the intervention and follow-up periods. Following treatment, VAS scores' analysis revealed beneficial effects on pain, vomiting, dyspnoea, fatigue, anorexia, insomnia, daily activity and psychological calmness. These improvements were maintained over the two-week follow-up phase. After the first Qi therapy session, the patient discontinued medication and could sit by himself; after the fourth session, the patient was able to walk and use the toilet without assistance (improvement in KPS: 70: care for self, unable to perform normal activity or to do active work). Although limited by the single case study approach, our results support previous studies on this topic and provide reasons to conduct controlled clinical trials. PMID- 16274469 TI - Conversion of the Caregiver Quality of Life Index to an interview instrument. PMID- 16274470 TI - Squamous cell carcinoma of colorectal cancer: adjuvant radiotherapy is necessary? PMID- 16274471 TI - Developing patient information: a UK approach. PMID- 16274473 TI - A non-parametric framework for estimating threshold limit values. AB - BACKGROUND: To estimate a threshold limit value for a compound known to have harmful health effects, an 'elbow' threshold model is usually applied. We are interested on non-parametric flexible alternatives. METHODS: We describe how a step function model fitted by isotonic regression can be used to estimate threshold limit values. This method returns a set of candidate locations, and we discuss two algorithms to select the threshold among them: the reduced isotonic regression and an algorithm considering the closed family of hypotheses. We assess the performance of these two alternative approaches under different scenarios in a simulation study. We illustrate the framework by analysing the data from a study conducted by the German Research Foundation aiming to set a threshold limit value in the exposure to total dust at workplace, as a causal agent for developing chronic bronchitis. RESULTS: In the paper we demonstrate the use and the properties of the proposed methodology along with the results from an application. The method appears to detect the threshold with satisfactory success. However, its performance can be compromised by the low power to reject the constant risk assumption when the true dose-response relationship is weak. CONCLUSION: The estimation of thresholds based on isotonic framework is conceptually simple and sufficiently powerful. Given that in threshold value estimation context there is not a gold standard method, the proposed model provides a useful non-parametric alternative to the standard approaches and can corroborate or challenge their findings. PMID- 16274472 TI - Mapping and characterization of the amplicon near APOA2 in 1q23 in human sarcomas by FISH and array CGH. AB - BACKGROUND: Amplification of the q21-q23 region on chromosome 1 is frequently found in sarcomas and a variety of other solid tumours. Previous analyses of sarcomas have indicated the presence of at least two separate amplicons within this region, one located in 1q21 and one located near the apolipoprotein A-II (APOA2) gene in 1q23. In this study we have mapped and characterized the amplicon in 1q23 in more detail. RESULTS: We have used fluorescence in situ hybridisation (FISH) and microarray-based comparative genomic hybridisation (array CGH) to map and define the borders of the amplicon in 10 sarcomas. A subregion of approximately 800 kb was identified as the core of the amplicon. The amplification patterns of nine possible candidate target genes located to this subregion were determined by Southern blot analysis. The genes activating transcription factor 6 (ATF6) and dual specificity phosphatase 12 (DUSP12) showed the highest level of amplification, and they were also shown to be over-expressed by quantitative real-time reverse transcription PCR (RT-PCR). In general, the level of expression reflected the level of amplification in the different tumours. DUSP12 was expressed significantly higher than ATF6 in a subset of the tumours. In addition, two genes known to be transcriptionally activated by ATF6, glucose-regulated protein 78 kDa and -94 kDa (GRP78 and GRP94), were shown to be over-expressed in the tumours that showed over-expression of ATF6. CONCLUSION: ATF6 and DUSP12 seem to be the most likely candidate target genes for the 1q23 amplification in sarcomas. Both genes have possible roles in promoting cell growth, which makes them interesting candidate targets. PMID- 16274474 TI - A flow cytometry based method for studying embryogenesis and immune reactivity to embryogenic stages in filarial parasites. AB - BACKGROUND: In the absence of intermediate animal hosts, the process of embryogenesis leading to fecundity of adult female filarial worms is very critical for persistence of these obligate parasites in human communities. Embryogenesis in adult female filarial parasites involves fertilization of eggs or oocytes by sperms and their subsequent development into motile microfilariae inside the uterine cavity of worms. Development of assays for monitoring embryogenesis in adult female worms is a critical requirement in filariasis research--filarial worms are known to harbour endosymbionts such as Wolbachia which play a significant role in fecundity. Tetracycline or doxycycline treatment of the infected hosts effectively eliminates the endosymbionts resulting in inhibition of embryogenesis in female worms. Currently, inhibition of embryogenesis in adult filarial worms can be monitored only by microscopic examination of in vitro harvested intrauterine stages. METHODS: Adult female filarial worms of bovine filarial parasites, Setaria digitata were collected from the peritoneum of infected animals and intrauterine stages were harvested in culture medium and were analyzed for forward and side scatter by flowcytometry using a BD FACS Calibur. Different populations were gated, sorted and identified by phase microscopy. Binding of biotinylated lectins to intra uterine stages was monitored using FITC labeled Avidin and monitored by flow cytometry of gated populations. Similarly, binding of antibodies in human filarial sera to intrauterine stages was monitored using FITC labeled anti-human immunoglobulins. RESULTS: The forward and side scatter for intrauterine stages delineated 3 distinct populations labeled as R1, R2 and R3. The three populations were sorted and identified to be a) fully stretched microfilariae, b) early and c) late developmental stages of eggs respectively. Lectins such as Wheat Germ agglutinin or Concanavalin-A were found to bind strongly to egg stages and less prominently to intra-uterine microfilariae. Similarly the binding of antibodies in filarial sera to the three intra-uterine stages could also be precisely quantified. CONCLUSION: The manuscript reports a novel flow cytometry based method to monitor progression of embryogenesis in adult filarial worms. Apart from relative quantification of different intra uterine developmental stages, the assay allows quantitative binding of lectins and antibodies to each of the intrauterine stages. It may now be possible to quantify levels of antibodies in infected and immune hosts to monitor anti-fecundity immunity in filariasis--the assay can thus be used as a powerful tool for drug development and in immunological studies in human and experimental filariasis. PMID- 16274475 TI - Increasing work-place healthiness with the probiotic Lactobacillus reuteri: a randomised, double-blind placebo-controlled study. AB - BACKGROUND: Short term illnesses, usually caused by respiratory or gastrointestinal diseases are disruptive to productivity and there is relatively little focus on preventative measures. This study examined the effect of the probiotic Lactobacillus reuteri protectis (ATCC55730) on its ability to improve work-place healthiness by reducing short term sick-leave caused by respiratory or gastrointestinal infections. METHODS: 262 employees at TetraPak in Sweden (day workers and three-shift-workers) that were healthy at study start were randomised in a double-blind fashion to receive either a daily dose of 108 Colony Forming Units of L. reuteri or placebo for 80 days. The study products were administered with a drinking straw. 181 subjects complied with the study protocol, 94 were randomised to receive L. reuteri and 87 received placebo. RESULTS: In the placebo group 26.4% reported sick-leave for the defined causes during the study as compared with 10.6% in the L. reuteri group (p < 0.01). The frequency of sick days was 0.9% in the placebo group and 0.4% in the L. reuteri group (p < 0.01). Among the 53 shift-workers, 33% in the placebo group reported sick during the study period as compared with none in the L. reuteri group(p < 0.005). PMID- 16274476 TI - Alternative splicing and protein function. AB - BACKGROUND: Alternative splicing is a major mechanism of generating protein diversity in higher eukaryotes. Although at least half, and probably more, of mammalian genes are alternatively spliced, it was not clear, whether the frequency of alternative splicing is the same in different functional categories. The problem is obscured by uneven coverage of genes by ESTs and a large number of artifacts in the EST data. RESULTS: We have developed a method that generates possible mRNA isoforms for human genes contained in the EDAS database, taking into account the effects of nonsense-mediated decay and translation initiation rules, and a procedure for offsetting the effects of uneven EST coverage. Then we computed the number of mRNA isoforms for genes from different functional categories. Genes encoding ribosomal proteins and genes in the category "Small GTPase-mediated signal transduction" tend to have fewer isoforms than the average, whereas the genes in the category "DNA replication and chromosome cycle" have more isoforms than the average. Genes encoding proteins involved in protein protein interactions tend to be alternatively spliced more often than genes encoding non-interacting proteins, although there is no significant difference in the number of isoforms of alternatively spliced genes. CONCLUSION: Filtering for functional isoforms satisfying biological constraints and accounting for uneven EST coverage allowed us to describe differences in alternative splicing of genes from different functional categories. The observations seem to be consistent with expectations based on current biological knowledge: less isoforms for ribosomal and signal transduction proteins, and more alternative splicing of interacting and cell cycle proteins. PMID- 16274477 TI - TOIB Study. Are topical or oral ibuprofen equally effective for the treatment of chronic knee pain presenting in primary care: a randomised controlled trial with patient preference study. [ISRCTN79353052]. AB - BACKGROUND: Many older people have chronic knee pain. Both topical and oral non- steroidal anti-inflammatory drugs (NSAIDs) are commonly used to treat this. Oral NSAIDS are effective, at least in the short term, but can have severe adverse effects. Topical NSAIDs also appear to be effective, at least in the short term. One might expect topical NSAIDs both to be less effective and to have fewer adverse effects than oral NSAIDs. If topical NSAIDs have fewer adverse effects this may outweigh both the reduction in effectiveness and the higher cost of topical compared to oral treatment. Patient preferences may influence the comparative effectiveness of drugs delivered via different routes. METHODS: TOIB is a randomised trial comparing topical and oral ibuprofen, with a parallel patient preference study. We are recruiting people aged 50 or over with chronic knee pain, from 27 MRC General Practice Research Framework practices across the UK. We are seeking to recruit 283 participants to the RCT and 379 to the PPS. Participants will be followed up for up to two years (with the majority reaching one year). Outcomes will be assessed by postal questionnaire, nurse examination, laboratory tests and medical record searches at one and two years or the end of the study. DISCUSSION: This study will provide new evidence on the overall costs and benefits of treating chronic knee pain with either oral or topical ibuprofen. The use of a patient preference design is unusual, but will allow us to explore how preference influences response to a medication. In addition, it will provide more information on adverse events. This study will provide evidence to inform primary care practitioners, and possibly influence practice. PMID- 16274478 TI - Identification of clustered microRNAs using an ab initio prediction method. AB - BACKGROUND: MicroRNAs (miRNAs) are endogenous 21 to 23-nucleotide RNA molecules that regulate protein-coding gene expression in plants and animals via the RNA interference pathway. Hundreds of them have been identified in the last five years and very recent works indicate that their total number is still larger. Therefore miRNAs gene discovery remains an important aspect of understanding this new and still widely unknown regulation mechanism. Bioinformatics approaches have proved to be very useful toward this goal by guiding the experimental investigations. RESULTS: In this work we describe our computational method for miRNA prediction and the results of its application to the discovery of novel mammalian miRNAs. We focus on genomic regions around already known miRNAs, in order to exploit the property that miRNAs are occasionally found in clusters. Starting with the known human, mouse and rat miRNAs we analyze 20 kb of flanking genomic regions for the presence of putative precursor miRNAs (pre-miRNAs). Each genome is analyzed separately, allowing us to study the species-specific identity and genome organization of miRNA loci. We only use cross-species comparisons to make conservative estimates of the number of novel miRNAs. Our ab initio method predicts between fifty and hundred novel pre-miRNAs for each of the considered species. Around 30% of these already have experimental support in a large set of cloned mammalian small RNAs. The validation rate among predicted cases that are conserved in at least one other species is higher, about 60%, and many of them have not been detected by prediction methods that used cross-species comparisons. A large fraction of the experimentally confirmed predictions correspond to an imprinted locus residing on chromosome 14 in human, 12 in mouse and 6 in rat. Our computational tool can be accessed on the world-wide-web. CONCLUSION: Our results show that the assumption that many miRNAs occur in clusters is fruitful for the discovery of novel miRNAs. Additionally we show that although the overall miRNA content in the observed clusters is very similar across the three considered species, the internal organization of the clusters changes in evolution. PMID- 16274480 TI - Effects of co-administered dexamethasone and diclofenac potassium on pain, swelling and trismus following third molar surgery. AB - BACKGROUND: The apparent interactions between the mechanisms of action of non steroidal anti-inflammatory drugs (NSAIDS) and steroids suggest that co-therapy may provide beneficial inflammatory and pain relief in the absence of side effects. The aim of the study was to compare the effect of co-administered dexamethasone and diclofenac potassium (diclofenac K) with diclofenac K alone on the postoperative pain, swelling and trismus after surgical removal of third molars. PATIENTS AND METHODS: A prospective randomized double-blind study was conducted at the Department of Oral and Maxillofacial Surgery, Lagos University Teaching Hospital, Nigeria. A total of 100 patients were randomly allocated to two treatment groups of dexamethasone (prophylactic 8 mg and postoperative 4 mg IV) and diclofenac K (50 mg Oral before and after surgery), and diclofenac K alone (as with first group). The overall analgesic efficacy of the drug combinations was assessed postoperatively by determination of pain intensity using a category rating scale. Facial swelling was measured using a tape measure placed from tragus to gonion to tragus, while interincisal mouth-opening of patients was measured using a vernier calibrated caliper pre-operatively and post operatively. RESULTS: Co-administration of dexamethasone and diclofenac K was significantly superior to diclofenac alone for the relief of pain (P < 0.05), and facial swelling up to post-operative 48 hour (P < 0.05). However, there was no significant difference for trismus relief between the two medication protocols (P > 0.05). CONCLUSION: This study illustrates enhanced effects of co-administered dexamethasone and diclofenac K on short-term post-operative pain and swelling, compared to diclofenac potassium alone in third molar surgery. PMID- 16274479 TI - Methylenetetrahydrofolate reductase polymorphism 677C>T is associated with peripheral arterial disease in type 2 diabetes. AB - BACKGROUND: Individuals with diabetes are twice as likely to develop peripheral arterial disease (PAD), the manifestation of extensive atherosclerosis throughout the lower extremities. One putative determinant of PAD is the 677C>T polymorphism in the gene encoding methylenetetrahydrofolate reductase (MTHFR), which has previously been found to associate with various diabetic complications including retinopathy, nephropathy, atherosclerosis and coronary heart disease. The objective of this study was to investigate a possible role for the MTHFR 677C>T gene polymorphism with PAD in subjects with type 2 diabetes from an isolated aboriginal Canadian population. METHODS: The 677C>T MTHFR gene polymorphism was genotyped in 138 subjects of Oji-Cree descent. Participants were selected from a community-wide survey that included PAD assessment by ankle-brachial index (ABI) measurement, and also intermittent claudication assessment by the Rose questionnaire. RESULTS: MTHFR 677T allele carriers had an increased risk of PAD with an odds ratio of 3.54 (95% CI 1.01, 12.4), P = 0.049, after adjustment for age, sex, duration of diabetes, hypertension, current smoking habits, and use of insulin or oral treatment for diabetes. None of these additional co-variables was significantly associated with PAD. No association was found between MTHFR genotype and intermittent claudication. CONCLUSION: The genetic influence of the MTHFR 677C>T genotype on diabetic PAD is modest, yet for the Oji-Cree it is a major risk factor in comparison to other traditional risk factors. PMID- 16274481 TI - Reasons for and consequences of missed appointments in general practice in the UK: questionnaire survey and prospective review of medical records. AB - BACKGROUND: Missed appointments are a common occurrence in primary care in the UK, yet little is known about the reasons for them, or the consequences of missing an appointment. This paper aims to determine the reasons for missed appointments and whether patients who miss an appointment subsequently consult their general practitioner (GP). Secondary aims are to compare psychological morbidity, and the previous appointments with GPs between subjects and a comparison group. METHODS: Postal questionnaire survey and prospective medical notes review of adult patients missing an appointment and the comparison group who attended appointments over a three week period in seven general practices in West Yorkshire. RESULTS: Of the 386 who missed appointments 122 (32%) responded. Of the 386 in the comparison group 223 (58%) responded, resulting in 23 case control matched pairs with complete data collection. Over 40% of individuals who missed an appointment and participated said that they forgot the appointment and a quarter said that they tried very hard to cancel the appointment or that it was at an inconvenient time. A fifth reported family commitments or being too ill to attend. Over 90% of the patients who missed an appointment subsequently consulted within three months and of these nearly 60% consulted for the stated problem that was going to be presented in the missed consultation. The odds of missing an appointment decreased with increasing age and were greater among those who had missed at least one appointment in the previous 12 months. However, estimates for comparisons between those who missed appointments and the comparison group were imprecise due to the low response rate. CONCLUSION: Patients who miss appointments tend to cite practice factors and their own forgetfulness as the main reasons for doing so, and most attend within three months of a missed appointment. This study highlights a number of implications for future research. More work needs to be done to engage people who miss appointments into research in a meaningful way. PMID- 16274483 TI - Increased PAI-1 plasma levels and risk of death from dengue: no association with the 4G/5G promoter polymorphism. AB - BACKGROUND: Dengue virus infected patients have high plasminogen activator inhibitor type I (PAI-1) plasma concentrations. Whether the insertion/deletion (4G/5G) polymorphism in the promotor region of the PAI-1 gene is associated with increased PAI-1 plasma concentrations and with death from dengue is unknown. We, therefore, investigated the relationship between the 4G/5G polymorphism and PAI-1 plasma concentrations in dengue patients and risk of death from dengue. METHODS: A total of 194 patients admitted to the Dr. Kariadi Hospital in Semarang, Indonesia, with clinical suspected severe dengue virus infection were enrolled. Blood samples were obtained on day of admission, days 1, 2 and 7 after admission and at a 1-month follow-up visit. Plasma concentrations of PAI-1 were measured using a sandwich ELISA kit. The PAI-1 4G/5G polymorphism was typed by allele specific PCR analysis. RESULTS: Concentrations of PAI-1 on admission and peak values of PAI-1 during admission were higher than the values measured in healthy controls. Survival was significantly worse in patients with PAI-1 concentrations in the highest tertile (at admission: OR 4.7 [95% CI 0.9-23.8], peak value during admission: OR 6.3 [95%CI 1.3-30.8]). No association was found between the PAI-1 4G/5G polymorphism, and PAI-1 plasma concentrations, dengue disease severity and mortality from dengue. CONCLUSION: These data suggest that the 4G/5G polymorphism has no significant influence on PAI-1 concentrations in dengue virus infected patients and is not associated with the risk of death from dengue. Other factors contributing to the variability of PAI-1 plasma concentrations in patients with dengue need to be explored. PMID- 16274482 TI - Evolution of subtype C HIV-1 Env in a slowly progressing Zambian infant. AB - BACKGROUND: Given the high prevalence of mother to child infection, the development of a better understanding of African subtype C HIV-1 transmission and natural evolution is of significant importance. In this study, we genotypically and phenotypically characterized subtype C viruses isolated over a 67-month follow-up period from an in utero-infected Zambian infant. Changes in genotype and phenotype were correlated to alterations of the host humoral immune response. RESULTS: A comparison of baseline maternal and infant samples indicated that the infant sequences are monophyletic and contain a fraction of the diversity observed in the mother. This finding suggests that selective transmission occurred from mother to child. Peaks in infant HIV-1 Env genetic diversity and divergence were noted at 48 months, but were not correlated with changes in co receptor usage or syncytia phenotype. Phylogenetic analyses revealed an accumulation of mutations over time, as well as the reappearance of ancestral lineages. In the infant C2-V4 region of Env, neither the median number of putative N-glycosylation sites or median sequence length showed consistent increases over time. The infant possessed neutralizing antibodies at birth, but these decreased in effectiveness or quantity with time. De novo humoral responses were detected in the child after 12 months, and corresponded with an increase in Env diversity. CONCLUSION: Our study demonstrates a correlation between HIV-1 Env evolution and the humoral immune response. There was an increase in genetic diversification in the infant viral sequences after 12 months, which coincided with increases in neutralizing antibody titers. In addition, episodes of viral growth and successive immune reactions in the first 5-6 years were observed in this slow progressor infant with delayed onset of AIDS. Whether this pattern is typical of slow progressing subtype C HIV-1 infected infant needs to be further substantiated. PMID- 16274484 TI - The pp24 phosphoprotein of Mason-Pfizer monkey virus contributes to viral genome packaging. AB - BACKGROUND: The Gag protein of Mason-Pfizer monkey virus, a betaretrovirus, contains a phosphoprotein that is cleaved into the Np24 protein and the phosphoprotein pp16/18 during virus maturation. Previous studies by Yasuda and Hunter (J. Virology. 1998. 72:4095-4103) have demonstrated that pp16/18 contains a viral late domain required for budding and that the Np24 protein plays a role during the virus life cycle since deletion of this N-terminal domain blocked virus replication. The function of the Np24 domain, however, is not known. RESULTS: Here we identify a region of basic residues (KKPKR) within the Np24 domain that is highly conserved among the phosphoproteins of various betaretroviruses. We show that this KKPKR motif is required for virus replication yet dispensable for procapsid assembly, membrane targeting, budding and release, particle maturation, or viral glycoprotein packaging. Additional experiments indicated that deletion of this motif reduced viral RNA packaging 6-8 fold and affected the transient association of Gag with nuclear pores. CONCLUSION: These results demonstrate that the Np24 domain plays an important role in RNA packaging and is in agreement with evidence that suggests that correct intracellular targeting of Gag to the nuclear compartment is an fundamental step in the retroviral life cycle. PMID- 16274485 TI - Sequential analysis of surfactant, lung function and inflammation in cystic fibrosis patients. AB - BACKGROUND: In a cross-sectional analysis of cystic fibrosis (CF) patients with mild lung disease, reduced surfactant activity was correlated to increased neutrophilic airway inflammation, but not to lung function. So far, longitudinal measurements of surfactant function in CF patients are lacking and it remains unclear how these alterations relate to the progression of airway inflammation as well as decline in pulmonary function over time. METHODS: As part of the BEAT trial, a longitudinal study to assess the course of airway inflammation in CF, we studied lung function, surfactant function and endobronchial inflammation using bronchoalveolar lavage fluid from 20 CF patients with normal pulmonary function (median FEV1 94% of predicted) at three times over a three year period. RESULTS: There was a progressive loss of surfactant function, assessed as minimal surface tension. The decline in surfactant function was negatively correlated to an increase in neutrophilic inflammation and a decrease in lung function, assessed by FEV1, MEF(75/25%VC), and MEF(25%VC). The concentrations of the surfactant specific proteins A, C and D did not change, whereas SP-B increased during this time period. CONCLUSION: Our findings suggest a link between loss of surfactant function driven by progressive airway inflammation and loss of small airway function in CF patients with limited lung disease. PMID- 16274486 TI - Hypothyroidism attenuates protein tyrosine nitration, oxidative stress and renal damage induced by ischemia and reperfusion: effect unrelated to antioxidant enzymes activities. AB - BACKGROUND: It has been established that hypothyroidism protects rats against renal ischemia and reperfusion (IR) oxidative damage. However, it is not clear if hypothyroidism is able to prevent protein tyrosine nitration, an index of nitrosative stress, induced by IR or if antioxidant enzymes have involved in this protective effect. In this work it was explored if hypothyroidism is able to prevent the increase in nitrosative and oxidative stress induced by IR. In addition the activity of the antioxidant enzymes catalase, glutathione peroxidase, and superoxide dismutase was studied. Control and thyroidectomized (HTX) rats were studied 24 h of reperfusion after 60 min ischemia. METHODS: Male Wistar rats weighing 380 +/- 22 g were subjected to surgical thyroidectomy. Rats were studied 15 days after surgery. Euthyroid sham-operated rats were used as controls (CT). Both groups of rats underwent a right kidney nephrectomy and suffered a 60 min left renal ischemia with 24 h of reperfusion. Rats were divided in four groups: CT, HTX, IR and HTX+IR. Rats were sacrificed and samples of plasma and kidney were obtained. Blood urea nitrogen (BUN) and creatinine were measured in blood plasma. Kidney damage was evaluated by histological analysis. Oxidative stress was measured by immunohistochemical localization of protein carbonyls and 4-hydroxy-2-nonenal modified proteins. The protein carbonyl content was measured using antibodies against dinitrophenol (DNP)-modified proteins. Nitrosative stress was measured by immunohistochemical analysis of 3 nitrotyrosine modified proteins. The activity of the antioxidant enzymes catalase, glutathione peroxidase, and superoxide dismutase was measured by spectrophotometric methods. Multiple comparisons were performed with ANOVA followed by Bonferroni t test. RESULTS: The histological damage and the rise in plasma creatinine and BUN induced by IR were significantly lower in HTX+IR group. The increase in protein carbonyls and in 3-nitrotyrosine and 4-hydroxy-2-nonenal modified proteins was prevented in HTX+IR group. IR-induced decrease in renal antioxidant enzymes was essentially not prevented by HTX in HTX+IR group. CONCLUSION: Hypothyroidism was able to prevent not only oxidative but also nitrosative stress induced by IR. In addition, the antioxidant enzymes catalase, glutathione peroxidase, and superoxide dismutase seem not to play a protective role in this experimental model. PMID- 16274487 TI - Lumbar segmental instability: a criterion-related validity study of manual therapy assessment. AB - BACKGROUND: Musculoskeletal physiotherapists routinely assess lumbar segmental motion during the clinical examination of a patient with low back pain. The validity of manual assessment of segmental motion has not, however, been adequately investigated. METHODS: In this prospective, multi-centre, pragmatic, diagnostic validity study, 138 consecutive patients with recurrent or chronic low back pain (R/CLBP) were recruited. Physiotherapists with post-graduate training in manual therapy performed passive accessory intervertebral motion tests (PAIVMs) and passive physiological intervertebral motion tests (PPIVMs). Consenting patients were referred for flexion-extension radiographs. Sagittal angular rotation and sagittal translation of each lumbar spinal motion segment was measured from these radiographs, and compared to a reference range derived from a study of 30 asymptomatic volunteers. Motion beyond two standard deviations from the reference mean was considered diagnostic of rotational lumbar segmental instability (LSI) and translational LSI. Accuracy and validity of the clinical assessments were expressed using sensitivity, specificity, and likelihood ratio statistics with 95% confidence intervals (CI). RESULTS: Only translation LSI was found to be significantly associated with R/CLBP (p < 0.05). PAIVMs were specific for the diagnosis of translation LSI (specificity 89%, CI 83-93%), but showed poor sensitivity (29%, CI 14-50%). A positive test results in a likelihood ratio (LR+) of 2.52 (95% CI 1.15-5.53). Flexion PPIVMs were highly specific for the diagnosis of translation LSI (specificity 99.5%; CI 97-100%), but showed very poor sensitivity (5%; CI 1-22%). Likelihood ratio statistics for flexion PPIVMs were not statistically significant. Extension PPIVMs performed better than flexion PPIVMs, with slightly higher sensitivity (16%; CI 6-38%) resulting in a likelihood ratio for a positive test of 7.1 (95% CI 1.7 to 29.2) for translation LSI. CONCLUSION: This study provides the first evidence reporting the concurrent validity of manual tests for the detection of abnormal sagittal planar motion. PAIVMs and PPIVMs are highly specific, but not sensitive, for the detection of translation LSI. Likelihood ratios resulting from positive test results were only moderate. This research indicates that manual clinical examination procedures have moderate validity for detecting segmental motion abnormality. PMID- 16274488 TI - Genetic covariance between indices of body condition and immunocompetence in a passerine bird. AB - BACKGROUND: Condition-dependence is a ubiquitous feature of animal life histories and has important implications for both natural and sexual selection. Mate choice, for instance, is typically based on condition-dependent signals. Theory predicts that one reason why condition-dependent signals may be special is that they allow females to scan for genes that confer high parasite resistance. Such explanations require a genetic link between immunocompetence and body condition, but existing evidence is limited to phenotypic associations. It remains unknown, therefore, whether females selecting males with good body condition simply obtain a healthy mate, or if they acquire genes for their offspring that confer high immunocompetence. RESULTS: Here we use a cross-foster experimental design to partition the phenotypic covariance in indices of body condition and immunocompetence into genetic, maternal and environmental effects in a passerine bird, the zebra finch Taeniopygia guttata. We show that there is significant positive additive genetic covariance between an index of body condition and an index of cell-mediated immune response. In this case, genetic variance in the index of immune response explained 56% of the additive genetic variance in the index of body condition. CONCLUSION: Our results suggest that, in the context of sexual selection, females that assess males on the basis of condition-dependent signals may gain genes that confer high immunocompetence for their offspring. More generally, a genetic correlation between indices of body condition and imuunocompetence supports the hypothesis that parasite resistance may be an important target of natural selection. Additional work is now required to test whether genetic covariance exists among other aspects of both condition and immunocompetence. PMID- 16274489 TI - Development of a health care policy characterisation model based on use of private health insurance. AB - OBJECTIVE: The aim of this study was to develop a policy characterisation process based on measuring shifts in use of private health insurance (PHI) immediately following implementation of changes in federal health care policy. METHOD: Population-based hospital morbidity data from 1980 to 2001 were used to produce trend lines in the annual proportions of public, privately insured and privately uninsured hospital separations in age-stratified subgroups. A policy characterisation model was developed using visual and statistical assessment of the trend lines associated with changes in federal health care policy. RESULTS: Of eight changes in federal health care policy, two (introduction of Medicare and Lifetime Health Cover) were directly associated with major changes in the trend lines; however, minor changes in trends were associated with several of the other federal policies. Three types of policy effects were characterised by our model: direction change, magnitude change and inhibition. Results from our model suggest that a policy of Lifetime Health Cover, with a sanction for late adoption of PHI, was immediately successful in changing the private: public mix. The desired effect of the 30% rebate was immediate only in the oldest age group (70+ years), however, introduction of the lifetime health cover and limitations in the model restricted the ability to determine whether or if the rebate had a delayed effect at younger ages. CONCLUSION: An outcome-based policy characterisation model is useful in evaluating immediate effects of changes in health care policy. PMID- 16274490 TI - Correlation between human papillomavirus infection and bladder transitional cell carcinoma. AB - BACKGROUND: To determine the association of human papillomavirus infection (HPV) and transitional cell carcinoma (TCC). METHODS: Using polymerase chain reaction, fifty-nine bladder tissue specimens of patients with transitional cell carcinoma of bladder compared with 20 bladder samples of cases with non-neoplastic disorders. RESULTS: Male to female ratio was similar in the two groups (50/9 vs. 16/4, P = 0.62). Mean age was 67 +/- 10.8 years and 52 +/- 20.3 years in the case and control groups, respectively (P = 0.6). Of the 59 tissue specimens with diagnosis of transitional cell carcinoma, HPV DNA was detected in 21 (35.6%) samples, while it was present in only one sample (5%) in the control group (P = 0.008). HPV18 was the most common type of virus with the incidence rate of 17/21(81%). CONCLUSION: HPV might play a causative role in transitional cell carcinoma of bladder in our geographic area. PMID- 16274493 TI - Vaccination and early protection against non-host-specific Salmonella serotypes in poultry: exploitation of innate immunity and microbial activity. AB - A recent European Union Directive required member states to put monitoring and control programmes in place, of which vaccination is a central component. Live Salmonella vaccines generally confer better protection than killed vaccines, because the former stimulate both cell-mediated and humoral immunity. Administering Salmonella bacteria orally to newly hatched chickens results in extensive gut colonization and a strong adaptive immune stimulus but broiler chickens are immunologically immature. However, colonization exerts a variety of rapid (within 24 h) protective effects. These include specific colonization inhibition (competitive exclusion) in which the protective bacteria exert a profound resistance to establishment and colonization by other related bacteria. This is thought to be primarily a metabolic attribute of the vaccinating bacteria but may also involve competition for attachment sites. The presence of large numbers of bacteria originating from a live Salmonella vaccine in the intestine can also induce infiltration of polymorphonuclear cells into the intestinal wall, which confers resistance to invasion and systemic spread by virulent Salmonella strains. This opens new perspectives for vaccine usage in broilers, layers and breeding poultry but also in other animals which show increased susceptibility to infection because of their young age or for other reasons, such as oral chemoprophylaxis or chemotherapy, where the lack of established normal gut flora is an issue. We recommend that all live vaccines considered for oral administration should be tested for their ability to induce the two protective effects described above. Further developments in live Salmonella vaccines are, however, currently hindered by fears associated with the use and release of live vaccines which may be genetically modified. PMID- 16274491 TI - Current molecular understanding of Axenfeld-Rieger syndrome. AB - Axenfeld-Rieger syndrome (ARS) is a rare autosomal dominant inherited disorder affecting the development of the eyes, teeth and abdomen. The syndrome is characterised by complete penetrance but variable expressivity. The ocular component of the ARS phenotype has acquired most clinical attention and has been dissected into a spectrum of developmental eye disorders, of which open-angle glaucoma represents the main challenge in terms of treatment. Mutations in several chromosomal loci have been implicated in ARS, including PITX2, FOXC1 and PAX6. Full-spectrum ARS is caused primarily by mutations in the PITX2 gene. The homeobox transcription factor PITX2 is produced as at least four different transcriptional and splicing isoforms, with different biological properties. Intriguingly, PITX2 is also involved in left-right polarity determination, although asymmetry defects are not a feature of ARS. In experimental animal models and in cell culture experiments using PITX2, abundant evidence indicates that a narrow window of expression level of this gene is vital for its correct function. PMID- 16274494 TI - Past mortality from infectious diseases and current burden of allergic diseases in England and Wales. AB - This study documents the changes in mortality in England and Wales over the last 100 years as a possible explanation for our increasingly allergy-prone Western society. A total of 53 million computerized recorded deaths, which occurred from 1901 to 2000 were analysed retrospectively. Childhood mortality decreased by 98%, from 40.6% of total annual deaths in 1901 to 0.9% in 2000. In 1901, 36.2% of all deaths and 51.5% of childhood deaths were from infectious diseases. By contrast in 2000, 11.6% of all deaths and only 7.4% of childhood deaths were from infectious diseases. Infectious diseases were a significant cause of childhood mortality in British cities until about 40 years ago. Several factors, including vaccination, antibiotics and improved sanitation have contributed to this trend. Survival of individuals with heightened immunity to infections may have led to natural selection of allergy-prone individuals in England and Wales. However, the relationship between changes in rates of infection and allergy is complex and not fully understood. PMID- 16274495 TI - The incidence of molluscum contagiosum, scabies and lichen planus. AB - We aimed to describe the incidence of new episodes of molluscum contagiosum, scabies and lichen planus presenting to general practitioners in England and Wales. We examined data collected in a sentinel practice network (the Weekly Returns Service of the Royal College of General Practitioners) in which about half a million persons were observed each year over the period 1994-2003. The incidence of molluscum contagiosum in males was 243/100,000 person-years and in females 231; of scabies, males 351, females 437; of lichen planus, males 32, females 37. Incidence varied by year and age. Ninety per cent of molluscum contagiosum episodes were reported in children aged 0-14 years, where incidence in 2000 (midpoint of a 6-year period of stable incidence) was 1265/100,000 (95% CI 1240-1290). Scabies affected all ages and annual incidence ranged between 233 (95% CI 220-246) in 2003 and 470 (95% CI 452-488) in 2000. Lichen planus occurred chiefly in persons aged over 45 years: incidence (all ages) ranged between 27 (95% CI 23-31) in 2003 and 43 (95% CI 37-49) in 1998. The relative risk of female to male incidence (all ages) of molluscum contagiosum was 0.95 (95% CI 0.91 0.99); of scabies 1.25 (95% CI 1.21-1.28); and of lichen planus 1.19 (95% CI 1.08 1.13). PMID- 16274496 TI - Maximizing statistical power in group-randomized vaccine trials. AB - Statistical power in group-randomized vaccine trials is complex: group randomization may increase power by capturing more transmission effects but may decrease power as more individuals are affected by a common source of variance. The former effect dominates when most infections arise from within the group and the latter when most arise outside. This process is complicated further when individuals possess partial natural immunity. Vaccine trials typically compare infection frequency (as measured by agent or antibody detection) in vaccinated vs. unvaccinated groups. To assess the relative contributions to statistical power by direct agent detection vs. serological detection of recent infection, we constructed stochastic compartmental models using differential equations describing all possible discrete states of the group. We contrasted models where natural immunity was absent, altered only the susceptible state, or altered both the susceptible and infected states. We observed the effects of endemic infection levels, the fraction of infection arising from outside the group, infectiousness vs. susceptibility vaccine effects and waning rates. There was significant enhancement of statistical power when serological testing separated infected and susceptible classes into subsets by natural immunity status. PMID- 16274497 TI - Matrix models for childhood infections: a Bayesian approach with applications to rubella and mumps. AB - Mathematical modelling is an established tool for planning and monitoring vaccination programmes. However, the matrices describing contact rates are based on subjective choices, which have a large impact on results. This paper reviews published models and obtains prior model probabilities based on publication frequency and expert opinion. Using serological survey data on rubella and mumps, Bayesian methods of model choice are applied to select the most plausible models. Estimates of the basic reproduction number R0 are derived, taking into account model uncertainty and individual heterogeneity in contact rates. Twenty-two models are documented, for which publication frequency and expert opinion are negatively correlated. Using the expert prior with individual heterogeneity, R0=6.1 [95% credible region (CR) 4.3-9.2] for rubella and R0=19.3 (95% CR 4.0 31.5) for mumps. The posterior modes are insensitive to the prior for rubella but not for mumps. Overall, assortative models with individual heterogeneity are recommended. PMID- 16274498 TI - Where diseases and networks collide: lessons to be learnt from a study of the 2001 foot-and-mouth disease epidemic. AB - This paper uses a graph-theoretical approach to investigate the properties of the observed network of disease transmission in the 2001 foot-and-mouth epidemic in the United Kingdom. This analysis revealed both global and local heterogeneity in the contact pattern between the infected premises in the first 3 weeks of the disease. In particular, the global heterogeneity contributed to the failure of the culling strategy imposed by the UK government. However, a more effective strategy targeting selective deletion of key premises in the network was not available once the epidemic had begun. We recommend that post-hoc analyses of this sort should become part of preventative and proactive policy rather than part of a reaction to an ongoing crisis. PMID- 16274499 TI - Spatial and temporal epidemiology of sporadic human cases of Escherichia coli O157 in Scotland, 1996-1999. AB - In Scotland, between 1995 and 2000 there were between 4 and 10 cases of illness per 100000 population per year identified as being caused by Escherichia coli O157, whereas in England and Wales there were between 1 and 2 cases per 100000 population per year. Within Scotland there is significant regional variation. A cluster of high rate areas was identified in the Northeast of Scotland and a cluster of low rate areas in central-west Scotland. Temporal trends follow a seasonal pattern whilst spatial effects appeared to be distant rather than local. The best-fit model identified a significant spatial trend with case rate increasing from West to East, and from South to North. No statistically significant spatial interaction term was found. In the models fitted, the cattle population density, the human population density, and the number of cattle per person were variously significant. The findings suggest that rural/urban exposures are important in sporadic infections. PMID- 16274500 TI - Differences in survival of Escherichia coli O157:H7 under various conditions that re-enact the cooking of lunches implicated in an outbreak of haemorrhagic diarrhoea. AB - Two elementary schools were served lunches that were cooked in the same kitchen. An outbreak of Escherichia coli O157:H7 occurred at one school where the dishes that were prepared for the school were lukewarm and kept for 33 min at an average temperature of 45 degrees C before serving. However, no outbreak occurred at the other school where dishes were hot and were kept for 60 min at an average temperature of 50 degrees C before serving. In a series of experiments on the survival of E. coli O157:H7 in the liquid portion of similarly prepared food, the population of E. coli O157:H7 was reduced by 10-3 by heating at 50 degrees C for 60 min and by only 10-1 by heating at 45 degrees C for 40 min. Further, E. coli O157:H7 survived at 45 degrees C for 40 min but not at 50 degrees C for 60 min at pH 4.0 with a 4.0% salt concentration that was similar to that of the liquid part of the food. These results indicate that pH and salt concentration of cooked food markedly affect the survival of E. coli O157:H7 and help to explain the occurrence of the disease outbreak at only one of the schools. PMID- 16274501 TI - Microbial health risk posed by table eggs in Trinidad. AB - A survey of the microbial quality of table eggs sold in Trinidad was conducted. For 23 poultry layer farms each visited twice approximately 1 month apart, 25 pooled eggs constituted a composite sample, for 14 shopping malls each visited twice approximately 1 month apart, six pooled eggs made a composite sample and for a total of 102 other retailers across the country each visited once over a 4 month period, six pooled eggs constituted a composite sample. Swabs of egg shells and egg content were tested for selected bacteria. Twenty-four (13.0%), 68 (37.0%), and two (1.1%) of a total of 184 composite eggs (shells, egg content or both) sampled were positive for Salmonella, Escherichia coli, and Campylobacter respectively. All 184 samples tested were negative for Listeria spp. Salmonella was recovered from seven (3.8%) egg shell samples only compared with 14 (7.6%) egg content samples only positive for the pathogen. Fifty-two (28.3%) egg shell samples and seven (3.8%) egg content samples were positive for E. coli. Both isolates of Campylobacter coli originated from egg contents. Of a total of 24 composite egg samples positive for Salmonella, eight different serotypes of Salmonella were isolated from a total of 24 Salmonella-positive composite eggs of which S. Enteritidis was the most prevalent, 58.3% (14/24). Salmonella Georgia was isolated for the first time in Trinidad. Failure to properly handle or heat table eggs sold in Trinidad poses a potential health hazard to consumers because of their poor microbial quality. PMID- 16274502 TI - An outbreak of norovirus gastroenteritis associated with wedding cakes. AB - We sought to determine the source of a norovirus outbreak among attendees of 46 weddings taking place during a single weekend. Norovirus-compatible illness was experienced by 332 (39%) of wedding guests surveyed; the outbreak affected up to 2700 persons. Illness was associated with eating wedding cake provided by a bakery common to the weddings (adjusted RR 4.5, P<0.001). A cake requiring direct hand contact during its preparation accounted for the majority of illness. At least two bakery employees experienced norovirus-compatible illness during the week preceding the weddings. Identical sequence types of norovirus were detected in stool specimens submitted by two wedding guests, a wedding hall employee, and one of the ill bakery employees. It is likely that one or more food workers at the bakery contaminated the wedding cakes through direct and indirect contact. These findings reinforce the necessity of proper food-handling practices and of policies that discourage food handlers from working while ill. PMID- 16274504 TI - Typhoid fever and typhoid hepatitis in Taiwan. AB - The annual incidence of typhoid fever in Taiwan was 2.1-3.6 cases per 1,000,000 population from 1995 to 2002. More than 80% of 45 patients with typhoid fever treated at National Taiwan University Hospital from 1996 to 2002 had elevated serum aminotransferase levels at presentation. Ten of these patients were treated during an outbreak in Taipei County in 2002, and seven of them who did not have pre-existing liver disease developed hepatitis, which was unrelated to other aetiologies. All Salmonella typhi isolates were susceptible to extended-spectrum cephalosporins and fluoroquinolones. Multidrug resistance (intermediate resistance to ampicillin, trimethoprim-sulphamethoxazole, and chloramphenicol) was found in one (2.5%) of the 40 isolates studied. Pulsed-field gel electrophoresis analysis demonstrated a high genetic diversity among S. typhi isolates and identified a novel clone associated with the 2002 outbreak. Physicians should be alert to the possibility of typhoid fever when patients, without other gastrointestinal symptoms, present with sustained fever and hepatitis. PMID- 16274503 TI - Sesame seed products contaminated with Salmonella: three outbreaks associated with tahini. AB - In November 2002, the first of three outbreaks of Salmonella Montevideo infection in Australia and New Zealand was identified in New South Wales, Australia. Affected persons were interviewed, and epidemiologically linked retail outlets inspected. Imported tahini was rapidly identified as the source of infection. The contaminated tahini was recalled and international alerts posted. A second outbreak was identified in Australia in June-July 2003 and another in New Zealand in August 2003. In a total of 68 S. Montevideo infections, 66 cases were contacted. Fifty-four (82%) reported consumption of sesame seed-based foods. Laboratory analyses demonstrated closely related PFGE patterns in the S. Montevideo isolates from human cases and sesame-based foods imported from two countries. On the basis of our investigations sesame-based products were sampled in other jurisdictions and three products in Canada and one in the United Kingdom were positive for Salmonella spp., demonstrating the value of international alerts when food products have a wide distribution and a long shelf life. A review of the controls for Salmonella spp. during the production of sesame-based products is recommended. PMID- 16274505 TI - The importance of Campylobacter coli in human campylobacteriosis: prevalence and genetic characterization. AB - The primary aim of this study was to detect and genotype Campylobacter coli strains from humans over a period of one year from November 2002 to October 2003. Campylobacter spp. were isolated from patients with symptoms of enteric infection. Amplified fragment length polymorphism (AFLP) analysis was used to identify the genetic diversity of C. coli strains by cluster analysis. A total of 18.6% of all Campylobacter isolates were identified as C. coli. These data show, that C. coli is indeed of importance to human campylobacter infections. Heterogeneous patterns were detectable among the human C. coli pool by AFLP analysis suggesting different sources of infection. A continuous seasonal shift of genotypes was detectable. PMID- 16274506 TI - Species diversity and antimicrobial resistance of Shigella spp. isolated between 2001 and 2004 from hospitalized children with diarrhoea in Kolkata (Calcutta), India. AB - The incidence, phenotypic characteristics and antimicrobial resistance patterns of 193 Shigella strains isolated from 2489 hospitalized children with acute diarrhoea were studied during January 2001 to August 2004. S. flexneri (60%) was the most prevalent serogroup, followed by S. sonnei (23.8%), S. dysenteriae (9.8%) and S. boydii (5.7%). Since 2002, S. flexneri 2a was the most dominant serotype. Almost all S. flexneri strains exhibited resistance to ampicillin, co trimoxazole, tetracycline, nalidixic acid and fluoroquinolones. After a lapse of almost 14 years, S. dysenteriae type 1 strains reemerged for the first time during 2002 and these strains were resistant to more than two antibiotics (multidrug resistance), including fluoroquinolones. An upsurge of similar resistance patterns was also noted among S. flexneri type 2a since December 2003. Resistance to fluoroquinolone increased year on year among S. dysenteriae type 1 and S. flexneri, but not in S. boydii or S. sonnei. Monitoring of antimicrobial susceptibility through a surveillance programme is recommended to select appropriate antibiotics for the effective treatment of shigellosis in this region. PMID- 16274507 TI - Molecular epidemiology of Legionella pneumophila environmental isolates representing nine different serogroups determined by automated ribotyping and pulsed-field gel electrophoresis. AB - The purposes of the study were (i) to describe the abundance and epidemiology of Legionellaceae in the man-made environment in a northern Italian area, (ii) to assess the concordance between pulsed-field gel electrophoresis (PFGE) and automated ribotyping (AR) techniques for genotyping L. pneumophila and (iii) to investigate the correlation between serogrouping and genotyping data. Water was sampled from reservoirs in 12 buildings across an area of 80-km radius. Despite the water temperature always being maintained above 55 degrees C, all of the buildings sampled were contaminated with Legionellaceae on at least one occasion and 63 L. pneumophila isolates representing nine different serogroups were collected. The two DNA methods revealed a high degree of genetic heterogeneity, even though identical L. pneumophila clones were recovered at different sites. The AR technique provided a fairly reliable approximation of PFGE results (73% concordance), however there was poor correlation between serogrouping and genotyping data as identical DNA fingerprints were shared by isolates of different serogroups. PMID- 16274508 TI - Group A streptococcal infections: trend and strain emm typing in an area of central Italy, 1985-2002. AB - A retrospective study of group A streptococcal (GAS) infections was performed for the period 1985-2002 in an area of central Italy. Although very severe diseases such as streptococcal toxic shock syndrome (STSS) were observed, a general increase in invasive infections was not found. Isolates of GAS were classified by M protein genotyping (emm typing) and analysed according to their origin from invasive and non-invasive infections. The predominant emm types were types 1, 4 and 12, followed by types 3, 6 and 28. During the study period the proportion of isolates of types 1 and 12 fell, while other types (3, 6, 22, 28 and 77) appeared. Isolates from invasive and non-invasive infections shared several emm types; however, most invasive strains belonged to five types only (types 1, 4, 12, 28 and 77), while non-invasive isolates were generally more heterogeneous. PMID- 16274509 TI - Haemophilus influenzae type b carriage among 3- to 24-month-old Turkish children. AB - There are few studies from developing countries on the epidemiology of Haemophilus influenzae (Hib) infections among infants and children. We set out to determine the prevalence of oropharyngeal Hib colonization among Turkish children younger than two years of age and to identify antimicrobial resistance among the isolates. A cross-sectional study was conducted on 818 healthy children and oropharyngeal secretions were sampled. The carriage rate of Hib was found to be 7.2% and this increased significantly with age. Carriage of Hib among 3- to 6 month-old children (3.5%) was higher than expected and was significantly higher among children who were passive smokers (P=0.04). Logistic regression analysis showed that breastfeeding status was the sole significant factor for colonization (OR 2.2, 95% CI 1.26-3.82). Antimicrobial susceptibility tests on 56 isolates of H. influenzae showed that 51.8% and 21.4% were resistant to trimethoprim sulphamethoxazole and ampicillin respectively. Other notable resistances were to cefalexin (10.7%) and chloramphenicol (3.6%); no isolates were resistant to ceftriaxone. PMID- 16274510 TI - Seroprevalence of SARS coronavirus antibody in household contacts. AB - Between March and July 2003, 671 cases of severe acute respiratory syndrome (SARS) were diagnosed in Taiwan with a total of 84 fatalities. After the epidemic, a serological survey was conducted involving the asymptomatic household contacts. Household contacts of 13 index patients were enrolled in the study. Contact history and clinical symptoms of the household contacts were recorded by standardized questionnaires. Blood samples of patients and household contacts were collected at least 28 days after symptom onset in the index patients or household exposure in the contacts for SARS-associated coronavirus (SARS-CoV) IgG testing. On the basis of this investigation, 29 persons (25 adults and 4 children) were identified as having had unprotected exposure to the index cases before infection-control practices were implemented. Laboratory evaluation of clinical specimens showed no evidence of transmission of SARS-CoV infection to any contacts. This investigation demonstrated that subclinical transmission among household contacts was low in the described setting. PMID- 16274511 TI - Investigation into a school enterovirus outbreak using PCR detection and serotype identification based on the 5' non-coding region. AB - A summer camp was followed by an outbreak of illness involving around 90 children. Investigations included individual questionnaires, inspection of the camp facilities, and laboratory analysis of water and clinical samples. Contamination of drinking and swimming water was demonstrated. An enterovirus was detected by polymerase chain reaction (PCR) and/or culture in 4/4 cerebrospinal fluid samples, 9/15 (60%) stool samples from symptomatic children and 2/9 (22%) stool samples from asymptomatic children. The virus was identified as an echovirus 3 by sequencing and phylogenetic analysis of a short 5' non-coding region (NCR) PCR product. Viruses from the outbreak clustered closely and an echovirus 3 from a temporally associated non-outbreak case could be readily distinguished. Despite the lack of a standardized approach, direct molecular detection and identification of enteroviruses is an efficient epidemiological tool. Here the 5'-NCR was successfully used for both detection and 'serotyping', and the close genetic relatedness of isolates was proven. PMID- 16274512 TI - Non-polio enterovirus isolation among families in Ulaanbaatar and Tov province, Mongolia: prevalence, intrafamilial spread, and risk factors for infection. AB - Studies of non-polio enterovirus prevalence and transmissibility in developing countries are limited and few studies have investigated specific risk factors for infection. An epidemiological survey of non-polio enterovirus among families in Mongolia was conducted in the late summer of 2003. Stools of 122 healthy persons were collected weekly for 5 weeks. Eight serotypes of non-polio enteroviruses (echovirus 30, 33, 12, 25, coxsackievirus A10, A2, A4, A24) were isolated from 62 persons, with an overall isolation rate of 51%, and 64% and 35% among children under 10 years and adults over age 21 years. Fifty-four per cent of isolations were due to intrafamilial infection. Analysis of risk factors for infection suggested contamination of indoor kitchen, bathroom, toilet, and waste disposal area. Hand washing after defecation was protective against infection. Our study findings stress the importance of hand washing and cleaning hygienic facilities to prevent infection by enteric viruses in the home environment. PMID- 16274513 TI - A controversial necrotizing enterocolitis outbreak in a neonatal unit. PMID- 16274514 TI - Tattoos, incarceration and hepatitis B and C among street-recruited injection drug users in New Mexico, USA: update. AB - To the Editor:In a previous report [1], we described significant risks for hepatitis B (HBV) and hepatitis C (HCV) positivity associated with receipt of tattoos, particularly while incarcerated, among a street-recruited population of injection drug users (IDUs) in New Mexico, United States from 1995 to 1997. Another recent report in this Journal, based on a study conducted on prisoners in Australia, found tattooing in prison to be an independent risk for HCV [2]. Another report also described a strong association between tattoos and HCV, but found the strongest association to be with commercial tattooing venues [3]. That study found the risk associated with receipt of tattoos in prison elevated, but not statistically significant. That same report reviewed other articles and found a significant risk for HCV infection associated with tattoos in six out of eight studies that had data available. Further, a recent U.S. Centers for Disease Control and Prevention (CDC) document summarized the literature on risks for hepatitis infections in correctional settings and developed extensive control guidelines [4]. PMID- 16274515 TI - Heuristics in education and clinical practice in urology. PMID- 16274516 TI - Development of prostate cancer quality indicators: a modified Delphi approach. AB - OBJECTIVES: There is evidence of variation in both the processes and outcomes of prostate cancer care, resulting in possible harm to patients and increased costs to the health system. Care could be improved by first identifying critical, measurable indicators that correlate with quality of care. This work was conducted to develop indicators of prostate cancer care using a modified three step Delphi approach. METHODS: A 17-member multidisciplinary panel reviewed potential indicators extracted from the medical literature through two consecutive rounds of rating followed by consensus discussion. The panel then prioritized the indicators selected in the previous two rounds. RESULTS: Of 31 possible indicators that emerged from 49 reviewed articles, 11 were prioritized by the panel as benchmarks for assessing the quality of surgical care for prostate cancer. The 11 indicators represent three levels of measurement (regional, hospital, individual provider) across several phases of care (diagnosis, surgery, pathology, and follow-up), as well as broad measures of outcomes. CONCLUSION: A systematic evidence- and consensus-based approach was used to develop quality indicators of prostate cancer care, with a focus on pre-, peri- and post-operative care as well as outcomes. Some of the indicators selected by the panel were also recommended by a similarly structured panel process. These indicators can be used by individual providers and organizations to monitor the quality of their services, and develop interventions to address any variations. PMID- 16274518 TI - The development of laparoscopic surgical skills in pediatric urologists: longterm outcome of a mentorship-training model. AB - INTRODUCTION AND OBJECTIVES: We previously reported the successful attainment of laparoscopic skills in a group of practicing pediatric urologists without previous formal laparoscopic training. During the mentorship period, the four urologists (trainees A, B, C, and D) performed a number of renal retroperitoneal laparoscopic procedures (RRLP) under the tutelage of an expert mentor. Specifically, trainee A performed or assisted in 8 RRLP while trainees B, C, and D performed/assisted in 10, 7, or 18 RRLP, respectively. Herein we assessed the outcome of this training program and practice pattern of this same group of urologists. METHODS: Following the completion of the mentorship period, we reviewed the outcomes of all of the consecutive RRLP performed from September 2001 to March 2005 with respect to operative time, conversion rate, perioperative complications and length of hospital stay (LOS). Furthermore, we attempted to correlate the number of procedures each surgeon performed both during and subsequent to the mentorship period. RESULTS: Fifty-two ablative RRLP including nephrectomy (n=38), partial nephrectomy (n=12), or synchronous bilateral nephrectomy (n=2), were performed on 50 patients (19 males, 31 females) with a mean age of 5.5 years (range 4 months-14 years). Trainee A performed 16/40 procedures, trainees B and C each performed 2/40, while trainee D performed 20/40 procedures. Mean operative time was 2.4 hours (range 1.5-6.3 hours). Five patients required open conversion due to inability to obtain retroperitoneal access (n=3) or failure to progress (n=2). Two patients (one nephrectomy, one partial nephrectomy) developed retroperitoneal urinomas requiring temporary urinary diversion. There were no other perioperative complications and mean LOS was 1.2 days (range 1-4 days). More advanced reconstructive procedures have since been performed with the aid of laparoscopic exposure; trainee D has thus far successfully performed 12 laparoscopically assisted pyeloplasties. CONCLUSIONS: This series demonstrates the effectiveness of the mentorship-training model to introduce RRLP to a pediatric urology training program. It is evident that the post-mentorship practice is affected by the number of cases initially performed during the training period. The development of an "expert" laparoscopist is dependent not only on initial training experience, but continued education through ongoing case exposure. PMID- 16274517 TI - French-Canadian linguistic validation of the NIH Chronic Prostatitis Symptom Index. AB - INTRODUCTION: The NIH Chronic Prostatitis Symptom Index (CPSI) is recommended in the clinical evaluation of men with chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS). However, its use is not possible in French speakers, as it has not been validated in this population. We performed a linguistic validation of the CPSI. METHODS: Linguistic translation followed the forward-backward forward technique and relied on professional medical translators, bilingual health professionals, and patient input. Along with the SF-12, the translated version was administered to a convenience sample of men presenting for pre vasectomy visits (controls) and to consecutive patients with established CP/CPPS (cases). Men with CP/CPPS were subsequently asked to complete a 14-day retest questionnaire. Psychometric testing addressed standard reliability and validity characteristics. RESULTS: Thirty-six cases and 38 controls with respective mean ages of 46.5 and 44.0 years participated and 33 (91.2%) cases completed the retest questionnaire. Pain (p<0.001), urinary (p<0.001) and quality-of-life (QOL) scale (p<0.001) score means differed between cases and controls. For the same scales, Cronbach's alphas for cases were respectively 0.70, 0.72 and 0.79 versus 0.80, 0.57, and 0.88 for controls. The retest product-moments were 0.83 for pain, 0.55 for urinary, and 0.83 for QOL scales. In cases, strong correlation was noted between QOL and pain scales (r=0.7), and between urinary and pain scales (r=0.6), versus moderate correlation between QOL and urinary scales (r=0.4). Negative correlation was recorded between CPSI scales and SF-12 scales, which ranged from 0.2 to -0.4. CONCLUSIONS: When applied to CPPS and control subjects, the French Canadian CPSI translation demonstrates excellent discriminant properties. Moreover, its reliability and validity characteristics confirm the qualities of the CPSI as a standard evaluative tool for men with CPPS. PMID- 16274520 TI - Renal medullary carcinoma as an incidental finding in a horseshoe kidney: case report and literature review. AB - Renal medullary carcinoma is rare and extremely aggressive neoplasm that typically affects young patients of African decent who demonstrate sickle cell trait or disease. Since the original description in 1995, only few cases have been reported outside the United States. A 29 year-old Canadian male of Afro Caribbean decent with sickle cell trait developed right-sided hemiparesis due to brain infarct. During the clinical work-up, a 3 cm renal tumor was detected in a horseshoe kidney. The patient died suddenly 2 weeks after the presentation of massive non-neoplastic pulmonary thromboembolism, confirmed at autopsy. The final diagnosis of renal medullary carcinoma was established after the autopsy. Due to the small size of the tumor and the limited metastatic spread only to the regional lymph nodes, the tumor was considered an incidental finding, and not the primary cause of patient's death. PMID- 16274519 TI - Urological manifestations of BK polyomavirus in renal transplant recipients. AB - OBJECTIVES: BK polyomavirus (BKV) disease in renal transplant recipients has become an increasingly problematic clinical entity. Complications of BKV disease lead to chronic allograft nephropathy and ultimately loss in greater than 50% of cases. We reviewed our experience with BKV disease over a 5-year period. PATIENTS AND METHODS: We performed 155 cadaveric and 168 live-related transplants between January 2000 and June 2005. During this period, seven patients had biopsy confirmed BKV disease, which compromised the renal function of six cadaveric and one live-related renal transplant recipients. BKV was suspected as a potential cause of renal function deterioration after eliminating other possibilities. BKV was then confirmed by detecting viral DNA in urine samples by polymerase chain reaction (PCR) and visualizing viral particles in allograft biopsies by electron microscopy. RESULTS: The deterioration of allograft function in five renal transplant recipients was due to polyomavirus-associated nephropathy and two due to ureteric stenosis. Upon confirmation of BKV, overall immunosuppression was reduced or modified with follow-up of 5 to 44 months. However, additional rescue therapies were used to stabilize allograft function including ciprofloxacin, intravenous immunoglobulin, and leflunomide. One patient died and another lost his allograft due to non-compliance and reverted to hemodialysis, but renal function in the remaining five allografts has remained stable at higher serum creatinine levels. CONCLUSIONS: The management of BKV disease in renal transplant recipients is not yet clearly defined. However, early recognition of urological sequelae and modification of immunosuppressive therapy are essential to ensure adequate long-term function of these allografts. PMID- 16274521 TI - Remission of hormone-refractory prostate cancer attributed to Essiac. AB - Essiac is a popular complementary and alternative medicine (CAM) that is utilized by many cancer patients in North America. Much anecdotal reporting exists about its cancer-fighting qualities, but so far no clinical trials have been preformed to validate those claims. We describe here the case of a 64-year-old man whose hormone-refractory prostate cancer responded well to Essiac tea. PMID- 16274522 TI - Specimen extraction with a surgical glove during laparoscopic radical prostatectomy. AB - INTRODUCTION AND OBJECTIVE: Cost containment during techniques of laparoscopic radical prostatectomy remains an ongoing concern. We present a cost-effective alternative technique of specimen extraction using a surgical glove. MATERIALS AND METHODS: We describe an alternative technique of prostate removal using a surgical glove in seven patients who underwent laparoscopic radical prostatectomy using a robotic-assisted technique. RESULTS: The glove extraction technique was successful in all seven patients for which the method was attempted without complication. CONCLUSION: Similar to commercially available devices, a surgical glove provides a reliable straightforward means of specimen extraction during laparoscopic radical prostatectomy. PMID- 16274523 TI - Pseudo-tumoral eosinophilic cystitis in a 3 year-old girl. AB - Eosinophilic cystitis (EC) is an uncommon form of bladder inflammation. It is a rare disorder in children and fewer than 25 cases have been described in the literature. We report a case of eosinophilic cystitis mimicking a bladder tumor in a 3 year-old girl with symptoms of urinary frequency. The diagnosis was confirmed with pathology and she underwent clinical treatment with corticosteroids. PMID- 16274524 TI - The core learning objectives education model: an approach to the teaching of core concepts in the clinical clerkship. AB - INTRODUCTION AND OBJECTIVE: The classical approach to the undergraduate medical clerkship has several limitations, including variability of clinical exposure and method of examination. As a result, the clerkship experience does not ensure exposure to and reinforcement of the fundamental concepts of a given specialty. MATERIALS AND METHODS: This article reviews the classic approach to clerkship education within the undergraduate medical education. Specific attention is placed on clinical exposure and clerkship examination. RESULTS: We describe the introduction of the Core Learning Objective (CLO) educational model at the University of Chicago Section of Urology. This model is designed to provide an efficient exposure to and evaluation of core clerkship learning objectives. CONCLUSIONS: The CLO model has been successfully initiated, focusing on both technical and clinical skill sets. The proposed model has been introduced with positive initial results and should allow for an efficient approach to the teaching and evaluation of core objectives in clerkship education. PMID- 16274525 TI - Metastatic umbilical mass from transitional cell carcinoma of the bladder. AB - We present the case of an 83-year-old female presenting with an umbilical mass measuring 2 cm x 3 cm. This patient had an extensive prior history of transitional cell carcinoma of the bladder. A fine needle aspiration of this umbilical mass confirmed this to be metastatic transitional cell carcinoma and the diagnostic workup demonstrated the presence of invasive bladder cancer with metastatic liver lesions. PMID- 16274529 TI - Passive standoff detection of SF6 at a distance of 5.7 km by differential Fourier transform infrared radiometry. AB - Recent results are presented on the passive detection, identification, and quantification of a vapor cloud of SF6 measured at a horizontal standoff distance of 5.7 km using a dual-beam interferometer optimized for background signal suppression. The measurements were performed at Defense Research and Development Canada (DRDC)-Valcartier during a number of recent open-air experiments. The measurement approach is based on the differential passive standoff detection method that has been developed by DRDC Valcartier during the past few years. This work represents the first such measurement reported in the open literature for a standoff distance as large as 5.7 km. These results clearly demonstrate the capability of the differential radiometry approach to the detection, identification, and quantification of chemical vapor clouds located at long distances from the sensor. PMID- 16274526 TI - Overeating after restraint stress in cholecystokinin-a receptor-deficient mice. AB - In mammals, including humans, a brain-gut hormone, cholecystokinin (CCK) mediates the satiety effect via CCK-A receptor (R). We generated CCK-AR gene-deficient (-/ ) mice and found that the daily food intake, energy expenditure, and gastric emptying of a liquid meal did not change compared with those of wild-type mice. Because CCK-AR(-/-) mice show anxiolytic status, we examined the effects of restraint stress. Seven hours of restraint stress was found to significantly decrease both body weight and food intake during the subsequent 3 days in all tested animals. On the fourth day after restraint stress, the CCK-AR(-/-) mice showed a significantly higher level of daily food intake than prior to stress, and food intake recovered to prestress levels in the wild-type mice. Since peripheral CCK-AR has been known to mediate gastric emptying, both gastric emptying and gastric acid secretion were determined to examine the mechanism of overeating in CCK-AR(-/-) mice. Neither gastric emptying nor gastric acid secretion differed between CCK-AR(-/-) and wild-type mice on the fourth day after stress. In contrast, however, the contents of dopamine and its metabolites in the cerebral cortex of CCK-AR(-/-) mice were increased by stress, but were rather decreased in wild-type mice. Changes in 5-hydroxytryptamine (5-HT) and its metabolite 5HIAA did not differ between the genotypes. In conclusion, CCK-AR(-/-) mice showed overeating after restraint stress, and dopaminergic hyperfunction in the brain of these mice was observed. The present evidence suggests that the CCK AR function, possibly via altering the dopaminergic function, might be involved in overeating after stress. PMID- 16274531 TI - Measurement of polystyrene nanospheres using excimer laser fragmentation fluorescence spectroscopy. AB - Monodisperse polystyrene nanospheres with a mean diameter of 102 nm are photofragmented with 193 nm light in N2 at laser fluences from 1 to 20 J/cm2. Carbon atom fluorescence at 248 nm from the disintegration of the particles is used as a signature of the polystyrene. The normalized fluorescence signals are self-similar with an exponential decay lifetime of approximately 10 ns. At fluences above 17 J/cm2, optical breakdown occurs and a strong continuum emission is generated that lasts significantly longer. A non-dimensional parameter, the photon-to-atom ratio (PAR), is used to interpret the laser-particle interaction energetics. Carbon fluorescence from polystyrene particles is compared with that from soot, and a similarity between the two particles is observed when normalized with PAR. Carbon emission from bulk polystyrene was also measured. Similar emission signals were observed, but the breakdown threshold of the surface is significantly lower at 0.2 J/cm2. PMID- 16274534 TI - Comparison of psychro-active arctic marine bacteria and common mesophillic bacteria using surface-enhanced Raman spectroscopy. AB - Psychro-active bacteria, important constituents of polar ecosystems, have a unique ability to remain active at temperatures below 0 degrees C, yet it is not known to what extent the composition of their outer cell surfaces aids in their low-temperature viability. In this study, aqueous suspensions of five strains of Arctic psychro-active marine bacteria (PAMB) (mostly sea-ice isolates), were characterized by surface-enhanced Raman spectroscopy (SERS) and compared with SERS spectra from E. coli and P. aerigunosa. We find the SERS spectra of the five psychro-active bacterial strains are similar within experimental reproducibility. However, these spectra are significantly different from the spectra of P. aeruginosa and E. coli. We find that the relative intensities of many of the common peaks show the largest differences reported so far for bacterial samples. An indication of a peak was found in the PAMB spectra that has been identified as characteristic of unsaturated fatty acids and suggests that the outer membranes of the PAMB may contain unsaturated fatty acids. We find that using suspensions of silver colloid particles greatly intensifies the Raman peaks and quenches the fluorescence from bacterial samples. This technique is useful for examination of specific biochemical differences among bacteria. PMID- 16274537 TI - Characterization of historic silk by polarized attenuated total reflectance Fourier transform infrared spectroscopy for informed conservation. AB - When assessing historic textiles and considering appropriate conservation, display, and storage strategies, characterizing the physical condition of the textiles is essential. Our work has concentrated on developing nondestructive or micro-destructive methodologies that will permit this. Previously, we have demonstrated a correlation between the physical deterioration of unweighted and "pink" tin (IV) chloride weighted silk and certain measurable spectroscopic and chromatographic signatures, derived from polarized Fourier transform infrared attenuated total reflectance (FTIR-ATR) spectroscopy (Pol-ATR) and high performance liquid chromatography (HPLC) microsampling analyses. The application of the Pol-ATR technique to aged silk characterization has now been extended to include a more comprehensive range of weighting methods and aging regimes. This was intended to replicate the full spectrum of states of deterioration observed in silk textiles, from pristine to heavily degraded. Breaking strength was employed as a measure of the physical integrity of the fibers, and, as expected, decreased with aging. An orientational crystallinity parameter, reflecting the microstructural ordering of the fibroin polymer within the fibers, was derived from the Pol-ATR spectra. A good correlation was observed between the breaking strength of the variety of fibers and this parameter. This suggests that the physical state of historic silk fabrics might be adequately characterized for conservation purposes by such indirect micromethodology. PMID- 16274542 TI - Spectral variable selection for partial least squares calibration applied to authentication and quantification of extra virgin olive oils using Fourier transform Raman spectroscopy. AB - The limits of quantitative multivariate assays for the analysis of extra virgin olive oil samples from various Greek sites adulterated by sunflower oil have been evaluated based on their Fourier transform (FT) Raman spectra. Different strategies for wavelength selection were tested for calculating optimal partial least squares (PLS) models. Compared to the full spectrum methods previously applied, the optimum standard error of prediction (SEP) for the sunflower oil concentrations in spiked olive oil samples could be significantly reduced. One efficient approach (PMMS, pair-wise minima and maxima selection) used a special variable selection strategy based on a pair-wise consideration of significant respective minima and maxima of PLS regression vectors, calculated for broad spectral intervals and a low number of PLS factors. PMMS provided robust calibration models with a small number of variables. On the other hand, the Tabu search strategy recently published (search process guided by restrictions leading to Tabu list) achieved lower SEP values but at the cost of extensive computing time when searching for a global minimum and less robust calibration models. Robustness was tested by using packages of ten and twenty randomly selected samples within cross-validation for calculating independent prediction values. The best SEP values for a one year's harvest with a total number of 66 Cretian samples were obtained by such spectral variable optimized PLS calibration models using leave-20-out cross-validation (values between 0.5 and 0.7% by weight). For the more complex population of olive oil samples from all over Greece (total number of 92 samples), results were between 0.7 and 0.9% by weight with a cross validation sample package size of 20. Notably, the calibration method with Tabu variable selection has been shown to be a valid chemometric approach by which a single model can be applied with a low SEP of 1.4% for olive oil samples across three different harvest years. PMID- 16274543 TI - A new Fourier transform infrared method for the determination of moisture in edible oils. AB - A rapid, practical, and accurate Fourier transform infrared (FT-IR) method for the determination of moisture content in edible oils has been developed based on the extraction of water from oil samples into dry acetonitrile. A calibration curve covering a moisture content range of 0-2000 ppm was developed by recording the mid-infrared (MIR) spectra of moisture standards, prepared by gravimetric addition of water to acetonitrile that had been dried over molecular sieves, in a 500 microm ZnSe transmission flow cell and ratioing these spectra against that of the dry acetonitrile. Water was measured in the resulting differential spectra using either the OH stretching (3629 cm(-1) or bending (1631 cm(-1)) bands to produce linear standard curves having standard deviations (SDs) of approximately +/-20 ppm. For moisture analysis in oils, the oil sample was mixed with dry acetonitrile in a 1:1 w/v ratio, and after centrifugation to separate the phases, the spectrum of the upper acetonitrile layer was collected and ratioed against the spectrum of the dry acetonitrile used for extraction. The method was validated by standard addition experiments with samples of various oil types, as well as with oil samples deliberately contaminated with alcohols, hydroperoxides, and free fatty acids to investigate possible interferences from minor constituents that may be present in oils and are potentially extractable into acetonitrile. The results of these experiments confirmed that the moisture content of edible oils can be assessed with high accuracy (on the order of +/-10 ppm) by this method, thus providing an alternative to the conventional, but problematic, Karl Fischer method and facilitating the routine analysis of edible oils for moisture content. PMID- 16274545 TI - Prescribing constraints in primary care: false economy or necessary husbandry? PMID- 16274546 TI - Using the Internet to promote sexual health awareness among young people. PMID- 16274547 TI - "Late deals, package care, it's really Hobson's choice.". PMID- 16274548 TI - Do sexual health services encourage teenagers to use condoms? A longitudinal study. AB - OBJECTIVE: Sexual health services have the potential to encourage teenagers' condom use through both the free supply of condoms and counselling. This study investigated whether 15/16-year-olds who attended sexual health services used condoms more and had different beliefs about condoms compared to those who did not use these services. METHODS: First, a cross-sectional multivariate model investigated the association between service visits and condom consistency (a ratio of the number of times a condom was used to the number of times a teenager had sexual intercourse in the past year) in teenagers at age 15/16 years (n = 1013). Second, a longitudinal multivariate model examined links between service use and changes in condom-related cognitions measured at age 13/14 and age 15/16 years (n = 3432). RESULTS: Visiting a service for free condoms was linked with greater condom consistency, after controlling for attitudes towards condoms, condom purchase and other factors. Visiting a service for other purposes was associated with lower consistency. Obtaining free condoms from services predicted greater condom self-efficacy and personal responsibility, and lower negative feelings relating to sexual pleasure when condoms were used. However, visiting a service for other purposes predicted less positive attitudes towards dual protection. CONCLUSIONS: Obtaining free condoms from services was associated with greater condom use and positive changes in attitudes towards condoms, although the role of service counselling remains unclear. Services could do more to stress the need for continued dual protection against sexually transmitted infections (STIs) when prescribing the pill. PMID- 16274549 TI - Involving service users in sexual health service development. AB - OBJECTIVES: The study objectives were to document users' experience of family planning and genitourinary medicine clinics and young people's services working within the time constraints of rapid service development and maximising the utility of this data for service improvement. METHODS: A total of 93 users of family planning and genitourinary medicine services participated in one of 13 facilitated discussion groups. Some 61% of the sample were women, 64% were aged over 25 years and 47% were Black Caribbean or Black African. The clinic journey was drawn on a wall covered with paper and participants added their comments during the discussion. RESULTS: Users had similar concerns across the three service types. Users perceived some receptionists and clinicians as un-friendly and judgmental and described others providing a quality service often under difficult conditions. Reception was insufficiently confidential, waiting environments uncomfortable, waiting times long and more information was needed throughout service use. CONCLUSIONS: Those elements of sexual health services known to be a source of dissatisfaction among young people may also be a problem for older service users and are experienced across different types of sexual health service. This preliminary study demonstrates the feasibility and acceptability of focus group evaluations of sexual health services. This approach generates qualitative data from relatively large numbers of users within a time scale consistent with service development. PMID- 16274551 TI - A qualitative study of women's use of emergency contraception. AB - BACKGROUND: While the use of emergency contraception (EC) is becoming more widespread in Australia, little is known about the reasons for, and the social context of, this use. METHODS: In order to explore the use of EC from the perspective of users, a qualitative study was conducted with women presenting to one of three health care settings in Melbourne, Australia for EC. RESULTS: Thirty two women ranging in age from 18 to 45 years were interviewed. While a number of themes were discussed with the women, this paper reports on four 'types of users' of EC identified from the data. 'Controllers' experienced failure of their contraceptive method and were very uncomfortable needing EC. They changed their contraceptive strategy in an attempt to avoid needing EC in the future. 'Thwarted controllers' were similar to controllers except that they could not improve their contraceptive strategy due to medical or social limitations. 'Risk takers' saw the use of EC as a component of their overall contraceptive strategy. They did not rely on EC regularly, but were comfortable to use it occasionally when the need arose. A final group of women were 'caught short' by a sexual experience that was unplanned and therefore they did not manage to use their chosen contraceptive strategy. CONCLUSIONS: The findings from this study challenge the assumptions that are often made about the users of EC and highlight the need to acknowledge the different ways that women make sense of, and make decisions about, contraception. PMID- 16274552 TI - Emergency contraception and prevention of induced abortion in India. AB - OBJECTIVE: Induced abortion is associated with high morbidity and mortality in India. Use of regular contraception with emergency contraception (EC) as backup can reduce the incidence of induced abortion. The study aimed to assess women's knowledge, practice, preference and acceptance of different contraceptive methods with special reference to the causes of induced abortion, and their willingness to use hormonal EC. METHODS: The study comprised a structured questionnaire survey conducted in the family planning clinic of a tertiary teaching hospital in New Delhi, India. A total of 623 women and three men seeking contraceptive advice and/or termination of pregnancy were interviewed. The main outcome measures were knowledge of different contraceptive methods including EC and the reasons for unintended pregnancy. RESULTS: More than 99% of the respondents knew about most of the modern methods of contraception whereas only 37 (5.9%; 95% CI 4.0-7.8) of the respondents knew about EC and none of them had ever used it. Contraceptive method failure led to unintended pregnancy in 39.1% (95% CI 33.7-44.5) of abortion seekers. Correct use of EC could have prevented nearly 65.5% (95% CI 57.0-74.0) of induced abortions due to contraceptive method failure and 25.6% (95% CI 20.7-30.5) of all induced abortions. CONCLUSIONS: More efforts are required to generate awareness about the safety, efficacy and availability of EC, regular use of effective contraception and the health hazards of induced abortion. PMID- 16274550 TI - Fear, hope and social desirability bias among women at high risk for HIV in West Africa. AB - BACKGROUND: Self-reports are widely used for measuring behaviour in HIV research and prevention, yet the accuracy of these measures has been shown to be questionable in many cases. Social desirability bias (SDB) is one of the key factors identified as affecting self-report accuracy. METHODS: Using in-depth interviews, we examined SDB from the perspective of 60 women at high risk for HIV in two West African countries: Ghana and Nigeria. We solicited suggestions for reducing SDB in the context of HIV research and prevention, and asked for feedback regarding methods currently being employed to reduce SDB. RESULTS: Themes pertaining to fear and a desire to have a better life were pervasive throughout the data. Thematic structure was similar between sites and age groups, although younger women tended to be more concerned about the interview context. CONCLUSIONS: Vulnerability of a population should be considered when asking sensitive questions. Audio-computer-assisted self-interviews may not be appropriate for vulnerable populations in developing countries, particularly for older respondents. PMID- 16274553 TI - Provision of emergency hormonal contraception through community pharmacies in a rural area. AB - OBJECTIVES: The provision of emergency hormonal contraception (EHC) through community pharmacies was introduced in Hambleton and Richmondshire, North Yorkshire, UK in December 2001 to contribute to the Teenage Pregnancy Strategy. The study aimed to establish how well the service is used, whether it is reaching the original target group, why people use the service and where it is accessed. METHODS: This was a descriptive study conducted in a rural primary care trust. RESULTS: From 1 January 2001 to 31 December 2003, there were 1412 pharmacy consultations for EHC and 1260 courses of EHC provided. General practitioner (GP) prescribing of EHC decreased but there was an overall increase in provision of EHC from pharmacies, GPs, family planning clinics, and accident and emergency departments. By December 2003, community pharmacies had become the largest provider of EHC. CONCLUSIONS: The supply of EHC through community pharmacies provided clients with wider choice and improved access to services, which resulted in increased overall provision of EC in this rural area. PMID- 16274554 TI - Chlamydia screening in general practice: views of professionals on the key elements of a successful programme. AB - OBJECTIVES: Chlamydia trachomatis is a common sexually transmitted infection with serious consequences if not treated. Chlamydia screening pilots in England have established feasibility in primary care but there are currently no examples of good practice in general practice. The objectives of the study were to understand issues of using general practice as a setting for chlamydia screening and to explore ways of implementing a successful screening strategy. METHODS: Based on findings of a literature review, a semi-structured schedule was constructed to interview a purposive sample of policymakers, consultants in sexual and reproductive health and primary care professionals. A thematic framework was used for qualitative analysis. RESULTS: Twenty-two themes were identified and were ranked in order of word count. The topic that generated most discussion was heterogeneity of knowledge, attitudes and skills in general practice. When broken down by professional group, this topic ranked the highest for practice nurses and consultants in sexual health; general practitioners (GPs) and the chlamydia screening coordinator spoke most about financial incentives while the public health consultant spoke most about access. CONCLUSIONS: Most believed screening can and should be done and general practice can offer better population coverage. It needs to have little impact on clinicians' workload, for example, by using urine tests and self-taken vaginal swabs. Financial recognition needs to reflect the administrative costs and the impact on reception staff, but this and the innovative tests might add to the cost of the screening programme. Incentives have to be handled sensitively to reduce inequity among GPs and other services offering screening. PMID- 16274556 TI - Are affluent, well-educated, career-orientated women knowledgeable users of the oral contraceptive pill? AB - OBJECTIVE: Studies have shown poor knowledge of oral contraceptives among women attending government health clinics and women in rural areas. Little is known about the level of contraceptive knowledge in educated, affluent, career orientated women, although it could be expected that access to information would be greater. The study objective was to describe the profile, knowledge and understanding of oral contraceptive users in a private general practice in Johannesburg, South Africa. METHODS: Over a period of 3 months, all women attending a private general practice who were using an oral contraceptive were asked to complete an anonymous questionnaire. Informed written consent was obtained in all cases. RESULTS: Fifty-one women participated in the study. Most women were nulliparous (71%), held a tertiary educational qualification (80%), were employed (84%) and were not concerned about the cost of their pill (65%). Most respondents (86%) obtained their information from a doctor. However, only 12% of women were aware of the danger of extending the active pill-free interval. Less than half (49%) were aware that their pill was less effective if taken more than 12 hours late and only 31% of women knew that their pill was effective again after taking seven active tablets. CONCLUSIONS: Educated, affluent women attending a private general practice lacked basic knowledge of the oral contraceptive pill. Consultations by practitioners need to be improved. PMID- 16274557 TI - An assessment of the first 3 years' use of Implanon in Luton. AB - OBJECTIVES: Implanon was introduced in the UK in September 1999. We present here the results of our first 106 Implanon insertions, performed over a period of 18 months. The aims of the study were to study the clinical and demographic profile of Implanon users, to assess the continuation rates of Implanon in the local population, and to identify the reasons for removal. METHODS: This was a case note-based study in which the data were transferred to a standardised pre-tested proforma. RESULTS: The age range of the 106 Implanon users was 15-43 years. Eighty-six of these clients had their Implanon removed and the Implanon status of 20 clients is not known since they were lost to follow-up by our service. Of these 86 cases, 26 had completed the full 3-year period; therefore, the continuation rate at 3 years was 30.2%. The continuation rate at the end of 1 year was 69.8% and at 2 years was 44.1%. Of the 60 women who had their Implanon removed before the recommended 3-year period, the most common reason was for bleeding irregularity (24 cases, 40%). CONCLUSIONS: This is the first published study set in the UK within a real-life setting to follow up a cohort of Implanon users for the full 3-year period. No contraceptive failures were found, replicating previous clinical trials. The continuation rate in this real-life situation was quite low compared to clinical trials. This is frequently the case when comparing real-life situations with clinical trials and may be in part due to higher motivation on the part of clinical trial participants. PMID- 16274558 TI - Terminology in early pregnancy loss: what women hear and what clinicians write. AB - OBJECTIVE: The Royal College of Obstetricians and Gynaecologists recommends that patient-focused terminology be used in early pregnancy care. The objective of the study was to evaluate whether inappropriate terminology is continuing to be used in Scottish gynaecology services. METHODS: A case note review (1259 records) and a patient survey (648 replies) assessed usage of four inappropriate terms ('abortion', 'blighted ovum', 'incompetent cervix' and 'pregnancy failure') in 18 Scottish hospitals providing secondary care to women with early pregnancy loss. RESULTS: Women reported hearing 'abortion' in 4.2% of hospital episodes (95% CI 2.9-6.0), but the term was used in 9.9% (95% CI 8.4-11.7) of hospital records. CONCLUSION: In order to meet national recommendations on terminology for early pregnancy loss, clinicians should not only say 'miscarriage' but also write it. PMID- 16274559 TI - Teenage pregnancies: is the high rate of teenage pregnancy and parenthood in the UK a public health problem? PMID- 16274562 TI - How to remove impalpable Implanon implants. PMID- 16274565 TI - Hidden agendas: psychosexual aspects in consultations. PMID- 16274567 TI - Workforce specialty review for family planning and reproductive health care 2003/2004: England, Wales, Northern Ireland and Scotland. PMID- 16274568 TI - Health needs of immigrants: rights to treatment and confidentiality. PMID- 16274569 TI - Thrombocytopenic purpura with depot medroxyprogesterone acetate and subsequent use of contraceptive implant: making decisions in the absence of evidence. PMID- 16274570 TI - Drug interactions with hormonal contraception. PMID- 16274572 TI - Confusion surrounding liver enzyme-inducing drugs. PMID- 16274574 TI - Use of contraception outside the terms of the product licence. PMID- 16274575 TI - Preoperative counselling for female sterilisation. PMID- 16274577 TI - Missed pill guidelines. PMID- 16274578 TI - Invisible contraception? PMID- 16274579 TI - Copying clinic letters to patients.